PMID- 19996973 TI - N-acetyltransferase 2, exposure to aromatic and heterocyclic amines, and receptor defined breast cancer. AB - The role of N-acetyltransferase 2 (NAT2) polymorphism in breast cancer is still unclear. We explored the associations between potential sources of exposure to aromatic and heterocyclic amines (AHA), acetylation status and receptor-defined breast cancer in 1020 incident cases and 1047 population controls of the German GENICA study. Acetylation status was assessed as slow or fast. Therefore, NAT2 haplotypes were estimated using genotype information from six NAT2 polymorphisms. Most probable haplotypes served as alleles for the deduction of NAT2 acetylation status. The risks of developing estrogen receptor alpha (ER) and progesterone receptor (PR)-positive or negative tumors were estimated for tobacco smoking, consumption of red meat, grilled food, coffee, and tea, as well as expert-rated occupational exposure to AHA with logistic regression conditional on age and adjusted for potential confounders. Joint effects of these factors and NAT2 acetylation status were investigated. Frequent consumption of grilled food and coffee showed higher risks in slow acetylators for receptor-negative tumors [grilled food: ER-: odds ratio (OR) 2.57, 95% confidence interval (CI) 1.07-6.14 for regular vs. rare; coffee: ER-: OR 2.55, 95% CI 1.22-5.33 for >or=4 vs. 0 cups/day]. We observed slightly higher risks for never smokers that are fast acetylators for receptor-positive tumors compared with slow acetylators (ER-: OR 1.32, 95% CI 1.00-1.73). Our results support differing risk patterns for receptor defined breast cancer. However, the modifying role of NAT2 for receptor-defined breast cancer is difficult to interpret in the light of complex mixtures of exposure to AHA. PMID- 19996974 TI - A meta-analysis on the utility of peripheral venous blood gas analyses in exacerbations of chronic obstructive pulmonary disease in the emergency department. AB - The objective of this case-based review is to identify and summarize the relevant evidence for the clinical utility of peripheral venous blood gas (pVBG) analyses in patients with acute exacerbations of chronic obstructive pulmonary disease (COPD) treated in the emergency department. Relevant studies were identified using the Cochrane Library, Medline, Embase, and CINAHL databases and by hand searching of references of published articles. Included studies were prospective trials comparing arterial and pVBG results in patients with COPD or respiratory distress that reported at least average differences and/or limits of agreement between the two results in English. Outcomes of interest were agreement between arterial and pVBG values for pH, pCO2, pO2, and HCO3. Eighty-nine studies were identified of which six were relevant. The weighted average difference for pCO2 was 5.92 mmHg, whereas those for pH, pO2, and HCO3 were 0.028, 18.65 mmHg, and 1.34 mEq/l, respectively. Using Bland-Altman analysis, the 95% limits of agreement were in the range of -0.10 to 0.08, -17 to 26 mmHg and, -3.5 to 3.5 mEq/l for pH, pCO2, and HCO3, respectively. Reported cutoff pVBG pCO2 values for screening of arterial hypercarbia ranged from 30 to 46 mmHg. No studies investigated the role of pVBG analysis in treatment alteration or clinical outcomes. Available evidence suggests that there is good agreement for pH and HCO3 values between arterial and pVBG results in patients with COPD, but not for pO2 or pCO2. Widespread clinical use is limited because of the lack of validation studies on clinical outcomes. PMID- 19996975 TI - Brugada-like ECG pattern induced by tricyclic antidepressants. AB - We present the case report of a 57-year-old woman with severe monointoxication with dosulepine (Prothiaden) who developed a Brugada-like electrocardiographic pattern. In tricyclic antidepressants (TCAs) poisoning the Brugada-like pattern on electrocardiogram is a characteristic albeit rare manifestation of the frequently occurring conduction abnormalities in the myocardium and its recognition is imperative as it is associated with a higher degree of morbidity and mortality. An overview of the literature is given and recommendations concerning treatment of TCA-induced arrhythmias are provided. After successful treatment, the electrocardiogram in the patient normalized. However, 4 days after intoxication, the ajmaline test was positive (pharmacological induction of a type I Brugada-like pattern), but a subsequent one, repeated after 11 days, was reportedly normal, probably because of the slow clearance of dosulepine. This raises questions about the specificity of ajmaline testing for Brugada syndrome in patients taking dosulepine and perhaps other TCAs and neuroleptic agents. PMID- 19996976 TI - Utilization of a UK emergency department by care home residents: a retrospective observational study. AB - There is little information available on the impact of care home residents upon emergency departments (EDs) in the UK. Our objective was to determine utilization of a UK ED by local care home residents. This study took place in a busy district general ED in an urban setting. Data were collected retrospectively for care home attendees during 2006. During the study period, an average of 2.4 care home patients attended per day. Just over half attended between 9 am and 5 pm. Fifty seven percent were admitted, of these 25% died. Most common presentation categories were trauma (38%), respiratory disorders (11.2%), cardiovascular disorders (10.8%), gastrointestinal disorders (9.3%) and genitourinary disorders (6.4%). Although accounting only for 1.5% of daily attendances, care home residents have a high admission and inpatient mortality rate. This study provides important information on presentation patterns for this subgroup of the population and identifies areas for further study. PMID- 19996977 TI - A placebo-controlled trial of 10-day bismuth-based quadruple therapy to eradicate Helicobacter pylori infection; a pilot study for the large Linqu County trial. AB - AIM: To explore a low-cost and highly-effective therapy for eradication of Helicobacter pylori (H. pylori), a placebo-controlled trial of quadruple therapy was conducted in a population at high risk of gastric cancer in Linqu County of Shandong Province, China. METHODS: Two hundred and seventy-seven adults aged 35 54 years with H. pylori infection in three villages were assigned to two groups: treatment (n=189 in two villages) and placebo (n=88 in one village). Participants received either a 10-day oral quadruple therapy regimen with omeprazole (20 mg, twice daily); tetracycline (750 mg, three times daily); metronidazole (500 mg, three times daily) and bismuth potassium citrate (300 mg, twice daily), or a similar lookalike placebo regimen. The status of H. pylori infection in each trial participant before and after six weeks of treatment was determined by a 13C urea breath test. RESULTS: One hundred and seventy-four of 189 participants completed the quadruple therapy (92.1%) and 84 participants completed the placebo therapy (95.5%). The H. pylori eradication rate by intention-to-treat analysis was 76.7% (145 of 189) in the treatment group and 1.1% (1 of 88) in the placebo group, respectively; by per-protocol analysis it was 83.3% (145 of 174) in the treatment group and 1.2% (1 of 84) in the placebo group, respectively. CONCLUSION: In a high-risk area of gastric cancer, we conducted a high compliance, tolerable, low side-effect and lowcost therapy of anti-H. pylori. The eradication rate of the 10-day quadruple treatment was more than 80% and significantly higher than the triple therapy regimen used in this population in an earlier trial. PMID- 19996978 TI - Efficacy of brief interventions in clinical care settings for persons living with HIV. AB - Prevention of HIV transmission from patients living with HIV (PLH) is a high national priority and strategies that are easy to implement and sustain to eliminate sexual transmission acts among PLH are needed. We evaluated a brief intervention that focused primarily on the enhancing motivations and encouraging PLH to act in accordance with their values without providing the intensity of the existing evidence-based programs for PLH. Using a quasiexperimental design, six medical clinics in Los Angeles County, CA, were evaluated across three intervention conditions: 1) computerized delivery; 2) provider delivery; or 3) standard care. We examined longitudinal changes in patients' reports of the number of HIV-negative (HIV-) or serostatus-unknown sexual partners and the number of unprotected vaginal and anal sex acts. Among 566 PLH, PLH in the computerized delivery condition reported a significant decrease in the number of HIV-/unknown sexual partners compared with the provider delivery and standard care conditions and a significant decrease in the number of unprotected sex acts in comparison to the standard care condition. Computerized motivational interventions delivered in waiting rooms at medical clinics may be an efficient strategy to reduce unprotected sex acts among PLH. PMID- 19996979 TI - An unusual case of left ventricular myxoma. PMID- 19996981 TI - Calcified anterior myocardial infarction depicted by cardiac MRI. AB - We present the case of a 68-year-old man with previous history of myocardial infarction and coronary artery bypass grafting. An initial chest X-ray showed a curvilinear calcification involving the left-sided profile of the mediastinal shadow. Cardiac MRI cine images demonstrated findings compatible with a calcified anterior myocardial infarction. PMID- 19996980 TI - Continuous positive airway pressure treatment in addition to optimal medical therapy for ventricular ectopy in a patient with heart failure and sleep-related breathing disorder. AB - In this report we describe the case of a 56-year-old man with a medical history of onset of asthenia, palpitations and dyspnoea for mild efforts. After a negative ergometric test for myocardial ischaemia and 24-h Holter monitoring showing frequent ventricular premature beats, but an echocardiogram with significant dilation and dysfunction of the left ventricle, coronary angiography was performed and did not show haemodynamically significant stenosis. Regarding the anamnesis of snoring, daytime sleepiness and the relationship between sleep related breathing disorder and cardiovascular disease, we performed a cardiorespiratory sleep study that indicated a diagnosis of sleep-related breathing disorder with prevalent obstructive apnoea. In addition to optimal medical therapy for cardiovascular disease, the patient began therapy with continuous positive airway pressure during the night. This showed a considerable decrease in ventricular premature beats (VPBs) during the night, a better control of ventricular ectopy during the day and a better compliance with medical therapy. PMID- 19996982 TI - Adipose tissue: a new source for cardiovascular repair. AB - Along with angiogenesis and gene therapy, stem cell transplantation is one of the newest treatment modalities proposed to improve the outcome of patients with heart failure or infarction. In this context, much interest has stemmed from experimental studies showing that cardiac transfer of unfractionated or partially purified bone marrow cells, or stem cells and progenitor cells derived from the bone marrow or peripheral blood, can enhance functional recovery after an acute myocardial infarction. On the basis of these data, stem cells and progenitor cells derived from the bone marrow have been proposed for use in the repair of cardiac tissue after acute myocardial infarction in patients. However, the relatively low abundance, small tissue volume, difficult accessibility and disease-related malfunction of bone marrow-derived stem cells make their clinical usefulness difficult in some situations. Recently it has been shown that adipose tissue contains a population of adult multipotent mesenchymal stem cells and endothelial progenitor cells that, in cell culture conditions, have extensive proliferative capacity and are able to differentiate into several lineages, including endothelial cells, smooth muscle cells and cardiomyocytes. The similarities between stem cells extracted from the bone marrow and the adipose tissue suggest the potential for the adipose tissue to act as an alternative, and perhaps preferable, cell source for repairing damaged tissues, such as the ischemic or infarcted heart. In this review we discuss molecular and functional characterization, preclinical results and currently ongoing clinical trials using adipose-derived stem cells in cardiovascular repair. PMID- 19996983 TI - A combination of rifaximin and neomycin is most effective in treating irritable bowel syndrome patients with methane on lactulose breath test. AB - AIM: There is a growing interest in methane and its association with constipation in functional bowel disease. Neomycin-based treatment of methane-positive subjects has resulted in improvement of constipation. Rifaximin, although superior for the treatment of irritable bowel syndrome compared with other antibiotics, seems less effective in methane-positive subjects. In this study, we evaluate 3 different antibiotic treatments in patients who have a methane positive breath test: rifaximin only, neomycin only, and the combination of neomycin and rifaximin. METHODS: A retrospective chart review was conducted on patients with methane on their lactulose breath test (> or =3 ppm of methane) who received one of the following antibiotic treatments: 500 mg b.i.d. for 10 days of neomycin alone, 400 mg t.i.d. for 10 days of rifaximin alone, or a combination of both rifaximin and neomycin for 10 days. All patients must have received antibiotic treatment after their initial consultation at the medical center and, in addition, had at least 1 follow-up to evaluate the effects of the treatment. After inclusion/exclusion criteria were met, all charts were evaluated to determine if the subject was a responder to the antibiotic therapy. This included clinical symptom improvement and eradication of methane on their breath test. RESULTS: Of the subjects receiving the treatment of rifaximin and neomycin (n=27), 85% had a clinical response, compared with 63% of subjects in the neomycin only group (n=8) (P=0.15) and 56% of subjects in the rifaximin only group (n=39) (P=0.01). When comparing the neomycin group with the rifaximin group, the difference was nonsignificant. When evaluating methane eradication results, 87% of subjects taking the rifaximin and neomycin combination eradicated the methane on their breath test. This is compared with 33% of subjects in the neomycin group that eradicated the methane (P=0.001), and only 28% of subjects in the rifaximin group (P=0.001). Of the patients who did not eliminate the methane with only rifaximin treatment, 66% of those who subsequently used the rifaximin and neomycin treatment were able to normalize their breath test. CONCLUSIONS: The combination of rifaximin and neomycin is more effective in treating methane producing subjects-in both clinical response and methane elimination. PMID- 19996985 TI - Marked variations in proximal colon cancer survival by race/ethnicity within the United States. AB - GOALS: To investigate the race/ethnicity and sex-specific variations in proximal colon cancer survival within the United States. BACKGROUND: Recent studies suggest a shift toward more proximal distributions of colon cancers. Survival benefits from screening colonoscopy may be limited to left-sided cancers. Identifying groups at greatest risk for mortality from proximal cancers will help to target health care resources to address these disparities. STUDY: A retrospective cohort study from 1973 to 2004 using a large population-based cancer registry to investigate demographic variations in colon cancer survival. RESULTS: Marked sex and race/ethnicity-specific variations in proximal colon cancer survival were observed. Overall 5-year survival was greatest in females (5 y survival, females: 44.5% and males: 41.7%, P<0.001) and in Asians (5-y survival, Asians: 50.4%, non-Hispanic whites: 43.1%, blacks: 39.7%, and Hispanics: 46.7%, P<0.001). Compared with females, males had significantly worse 5-year survival outcomes [odds ratios (OR), 0.77; 95% confidence intervals (CI), 0.75-0.79]. Compared with non-Hispanic whites, blacks had worse 5-year survival odds (OR, 0.74; 95% CI, 0.70-0.78) and Asians had better survival outcomes (OR, 1.16; 95% CI, 1.08-1.24). CONCLUSIONS: There exists significant sex and race/ethnicity-specific disparities in 5-year survival from proximal colon cancer. These differences may be explained by variations in delivery of health care between demographic groups or differences in disease biology specific to each group. PMID- 19996984 TI - A randomized, double-blind, phase 3 study of fospropofol disodium for sedation during colonoscopy. AB - GOALS: This double-blind, multicenter study evaluated the safety and efficacy of intravenous fospropofol (6.5 mg/kg vs. 2 mg/kg) for moderate sedation in patients undergoing colonoscopy. METHODS: In all, 314 patients >or=18 years (American Society of Anesthesiologists PS1 to PS3) were randomized to receive fospropofol 2 mg/kg, fospropofol 6.5- mg/kg, or midazolam 0.02 mg/kg, after pretreatment with intravenous fentanyl 50 mcg. Supplemental doses of study medication were permitted to achieve a Modified Observer's Assessment of Alertness/Sedation scale score 10 h x wk(-1) riding in a car or >23 h x wk(-1) of combined sedentary behavior had 82% and 64% greater risk of dying from CVD than those who reported <4 or <11 h x wk(-1), respectively. The pattern of the association did not materially change after multivariate adjustment. Regardless of the amount of sedentary activity reported by these men, being older, having normal weight, being normotensive, and being physically active were associated with a reduced risk of CVD death. CONCLUSION: In men, riding in a car and combined time spent in these two sedentary behaviors were significant CVD mortality predictors. In addition, high levels of physical activity were related to notably lower rates of CVD death even in the presence of high levels of sedentary behavior. Health promotion efforts targeting physically inactive men should emphasize both reducing sedentary activity and increasing regular physical activity for optimal cardiovascular health. PMID- 19996996 TI - Dose-response relationship of resistance training in older adults: a meta analysis. AB - PURPOSE: The purpose of this study was to determine the dose-response relationship of resistance training (RT) to improve strength and function in older adults. METHODS: A systematic literature search was performed in relevant databases and study reference lists to identify randomized controlled trials. Randomized controlled trials comparing the effects of different doses of strength training in older people (65 yr and older) on strength and functional outcomes were eligible. Two independent reviewers decided on study inclusion, extracted data, and assessed methodological quality. Standardized mean difference (SMD) and 95% confidence intervals (CI) were calculated for relevant outcomes and pooled using a random-effects model. RESULTS: Twenty-nine trials with a total of 1313 subjects (mean age = 65-81 yr) are summarized in this review. Trials comparing different training intensities show strong effects of progressive resistance training (PRT) on maximal strength in a dose-dependent manner, with high intensity (HI) PRT being more effective compared with moderate (MI)- and low intensity (LI) PRT (SMD [HI vs LI] = 0.88, 95% CI = 0.21-1.55; SMD [HI vs MI] = 0.62, 95% CI = 0.22-1.03). PRT was also successful for improving functional outcomes, but gains were independent of training intensity. Power training (PT) was more effective for improving muscle power (SMD [PT vs PRT] = 1.66, 95% CI = 0.08-3.24) and functional outcomes than PRT. There was only little information available on training volume and frequency. DISCUSSION: Higher training intensities are superior to lower intensities for improving maximal strength but not necessarily for functional performance of older adults. PT has shown to be a particularly effective method for enhancing muscle power and functional performance. More research is necessary to identify the effect of different training volumes and frequencies and the dose-response relationship for very old and frail populations. PMID- 19996997 TI - Epoch length and its effect on physical activity intensity. AB - BACKGROUND: Researchers have begun to investigate the issue of epoch length on children's physical activity using small sample sizes, and to date, no studies have been conducted in adolescents. PURPOSE: The purpose of this study was to investigate the effect of different epoch lengths (5, 15, 30, and 60 s) on derived levels of physical activity in both a child and an adolescent sample. METHODS: Three hundred and eleven children age 7-11 yr and 234 adolescents age 12 16 yr were asked to wear an accelerometer during waking hours for 7 d. The epoch was set at 5 s, and when data were downloaded, the activity counts were then reintegrated into 15-, 30-, and 60-s epochs. RESULTS: A significant epoch effect was seen for time spent in vigorous physical activity, light physical activity, and rest in the child and adolescent samples and moderate-to-vigorous and moderate physical activities in the child sample only. The Bland-Altman analysis showed reasonable agreement on moderate-to-vigorous, moderate, vigorous, and light physical activities and rest between 5- and 15-, 5- and 30-, and 30- and 60 s epochs in the child sample and between 5- and 15-, 5- and 30-, and 15- and 30-s epochs in the adolescent sample. CONCLUSIONS: A short epoch is strongly recommended for child and adolescent samples to obtain a "real" picture of young people's physical activity behavior and to prevent accumulation of counts reflecting the average activity level when longer epochs are used. Activity prevalence studies using epoch lengths of 5 and 60 s in a child or an adolescent sample should not be compared nor should 15- and 60- and 30- and 60-s epochs in an adolescent sample. PMID- 19996998 TI - Training effects on skeletal muscle calcium handling in human chronic heart failure. AB - PURPOSE: Patients with chronic heart failure (CHF) typically complain about skeletal muscle fatigue. In rat experiments, reduced intracellular calcium release seems to be related to fatigue development in normal skeletal muscle but not in muscle from rats with CHF. We therefore hypothesize that training may not improve intracellular calcium cycling to the same extent in muscles from patients with CHF compared with healthy controls (HC). METHODS: Thirteen HC and 11 CHF patients performed 6 wk of unilateral knee extensor endurance training. Computed tomographic examinations of the thigh and biopsies of vastus lateralis were obtained bilaterally before and after the training period. RESULTS: Peak power of the trained leg was 10% and 14% greater than that in the untrained leg in HC and CHF, respectively. For the HC, training resulted in a higher Ca2+ release rate and a lower leak in the trained leg associated with a tendency of increased ryanodine receptor (RyR) content with reduced phosphorylation level. In the trained leg of CHF patients, RyR content was reduced without associated changes of either Ca2+ leak or release rate. CONCLUSIONS: Training in HC has an effect on Ca2+ leak and release of the sarcoplasmic reticulum, but in CHF patients, training is achieved without such changes. Thus, calcium handling seems not to be the site of decreased exercise tolerance in CHF. PMID- 19996999 TI - Pressure pain sensitivity mapping in experimentally induced lateral epicondylalgia. AB - PURPOSE: The aim of this study was to apply topographical techniques to investigate changes in pressure pain sensitivity after induction of delayed onset muscle soreness (DOMS) in the elbow region in healthy subjects. METHODS: Pressure pain thresholds (PPT) were assessed over 12 points forming a 3 x 4 matrix (4 points in the superior part, 4 points in the middle, and 4 points in the lower part around the lateral epicondyle) over the dominant elbow in 13 healthy men, and pressure sensitivity maps were calculated. DOMS, as a model for lateral epicondylalgia (LE), was induced by repetitive high-level eccentric contractions of the wrist extensor muscles. PPT maps were assessed before, immediately after, and 24 h after eccentric exercise (DOMS). RESULTS: The two-way repeated-measure ANOVA detected significant differences in mean PPT for the measurement points (F = 5.96, P < 0.001), with lower PPT over the points located over the extensor carpi radialis brevis muscle. There was also a significant effect of time (F = 121.3, P < 0.001) but no time x point location interaction (F = 0.7, P = 0.8). PPT were lower 24 h after (P < 0.001) but not immediately after (P > 0.05) eccentric exercise. CONCLUSIONS: The study provides new key information regarding mechanical pain hyperalgesia in experimentally induced LE. Topographical pressure pain sensitivity maps from the elbow region revealed heterogeneously distributed mechanical sensitivity before and during DOMS. The most sensitive localizations for PPT assessment correspond to the muscle belly of the extensor carpi radialis brevis. Our results support the implication of the extensor carpi radialis brevis muscle in LE. PMID- 19997000 TI - Comparison of the ActiGraph 7164 and the ActiGraph GT1M during self-paced locomotion. AB - PURPOSE: This study compared the ActiGraph accelerometer model 7164 (AM1) with the ActiGraph GT1M (AM2) during self-paced locomotion. METHODS: Participants (n = 116, aged 18-73 yr, mean body mass index = 26.1 kg x m(-2)) walked at self selected slow, medium, and fast speeds around an indoor circular hallway (0.47 km). Both activity monitors were attached to a belt secured to the hip and simultaneously collected data in 60-s epochs. To compare differences between monitors, the average difference (bias) in count output and steps output was computed at each speed. Time spent in different activity intensities (light, moderate, and vigorous) based on the cut points of Freedson et al. was compared for each minute. RESULTS: The mean +/- SD walking speed was 0.7 +/- 0.22 m x s( 1) for the slow speed, 1.3 +/- 0.17 m x s(-1) for medium, and 2.1 +/- 0.61 m x s( 1) for fast speeds. Ninety-five percent confidence intervals (95% CI) were used to determine significance. Across all speeds, step output was significantly higher for the AM1 (bias = 19.8%, 95% CI = -23.2% to -16.4%) because of the large differences in step output at slow speed. The count output from AM2 was a significantly higher (2.7%, 95% CI = 0.8%-4.7%) than that from AM1. Overall, 96.1% of the minutes were classified into the same MET intensity category by both monitors. CONCLUSIONS: The step output between models was not comparable at slow speeds, and comparisons of step data collected with both models should be interpreted with caution. The count output from AM2 was slightly but significantly higher than that from AM1 during the self-paced locomotion, but this difference did not result in meaningful differences in activity intensity classifications. Thus, data collected with AM1 should be comparable to AM2 across studies for estimating habitual activity levels. PMID- 19997001 TI - ACE and AGTR1 polymorphisms and left ventricular hypertrophy in endurance athletes. AB - OBJECTIVE: This study aimed to evaluate the role of angiotensin type 1 receptor gene (AGTR1) polymorphism (A1166C) in left ventricular hypertrophy (LVH) mediated by the angiotensin-converting enzyme (ACE) in endurance athletes. METHODS: A group of 74 white, healthy male endurance athletes, aged between 25 and 40 yr, were enrolled in this study. All of them participated primarily in isotonic sports, training for at least >10 h x wk(-1), for at least 5 yr. The ACE genotype (insertion [I] or deletion [D] alleles) was ascertained by polymerase chain reaction (DD in 35, ID in 36, and II in 3). Group II was excluded from the analysis because of its small size. No difference was found between the two groups as regards age, blood pressure, HR, and echocardiographic data. RESULTS: The left ventricular mass index (LVMI) was significantly higher in group DD rather than in group ID (P = 0.029). The group DD showed a slightly higher prevalence of subjects with LVH (LVMI > 131 g x m(-2); 62.9%) than group ID (44.4%, P = 0.120). No association was found between ACE-DD and LVH (odds ratio (OR) = 2.12, 95% confidence interval = 0.82-5.46). Concerning the role of AGTR1 polymorphism, the highest LVMI was found in 15 athletes with ACE-DD and AGTR1 AC/CC genotypes (150 +/- 23 g x m(-2)); the lowest value of LVMI was found in the case of ACE-ID and AGTR1-AA (127 g x m(-2) +/- 18 g x m(-2)), whereas LVMI in subjects with ACE-DD + AGTR1-AA was similar to that in the ACE-ID + AGTR1-AC/CC group (134 +/- 18 g x m(-2) vs 133 +/- 20 g x m(-2), P = 0.880). The presence of ACE-DD + AGTR1 + AC/CC was strongly associated with LVH (OR = 4.6, P = 0.029). Moreover, subjects with LVH showed longer left ventricular isovolumetric relaxation time and higher end-systolic wall stress. The latter was strongly correlated to LVMI (r = 0.588), especially in the presence of ACE-DD + AGTR1 + AC/CC (r = 0.728). CONCLUSIONS: LVMI may be greater in the presence of ACE- DD and AGTR1-AC/CC polymorphisms. PMID- 19997002 TI - Branched-chain amino acid ingestion can ameliorate soreness from eccentric exercise. AB - PURPOSE: The purpose of this study was to examine the role of branched-chain amino acid (BCAA) supplementation during recovery from intense eccentric exercise. METHODS: Twenty-four non-weight-trained males were assigned to one of two groups: one group (supplementary, SUP) ingested BCAA beverages (n = 12); the second group (placebo, PLA) ingested artificially flavored water (n = 12). Diet was controlled throughout the testing period to match habitual intake. The eccentric exercise protocol consisted of 12 x 10 repetitions of unilateral eccentric knee extension exercise at 120% concentric one repetition maximum. On the day of the exercise, supplements were consumed 30 min before exercise, 1.5 h after exercise, between lunch and dinner, and before bed. On the following 2 d, four supplements were consumed between meals. Muscle soreness, muscle function, and putative blood markers of muscle damage were assessed before and after (1, 8, 24, 48, and 72 h) exercise. RESULTS: Muscle function decreased after the eccentric exercise (P < 0.0001), but the degree of force loss was unaffected by BCAA ingestion (51% +/- 3% with SUP vs -48% +/- 7% with PLA). A decrease in flexed muscle soreness was observed in SUP compared with PLA at 48 h (21 +/- 3 mm vs 32 +/- 3 mm, P = 0.02) and 72 h (17 +/- 3 mm vs 27 +/- 4 mm, P = 0.038). Flexed muscle soreness, expressed as area under the curve, was lower in SUP than in PLA (P = 0.024). CONCLUSIONS: BCAA supplementation may attenuate muscle soreness, but it does not ameliorate eccentric exercise-induced decrements in muscle function or increases in reputed blood markers of muscle damage, when consumed before exercise and for 3 d after an eccentric exercise bout. PMID- 19997003 TI - Timing protein intake increases energy expenditure 24 h after resistance training. AB - PURPOSE: To determine whether protein supplementation (PRO) before an acute bout of heavy resistance training (HRT) would influence postexercise resting energy expenditure (REE) and the nonprotein respiratory exchange ratio (RER). HYPOTHESIS: REE would be increased and RER would be decreased up to 48 h after timed PRO and HRT compared with CHO supplementation and HRT. METHODS: Eight resistance-trained subjects (five men and three women) participated in a double blind two-trial crossover design, where REE and RER were measured (7:00 a.m.) on four consecutive days. On the second day of trial 1, subjects consumed 376 kJ of either PRO (18 g of whey protein, 2 g of carbohydrate, 1.5 g of fat) or CHO (1 g of whey protein, 19 g of carbohydrate, 1 g of fat) 20 min before a single bout of HRT (nine exercises, 4 sets, 70%-75% 1-repetition maximum). REE and RER were measured 24 and 48 h after HRT. During trial 2, the same protocol was followed except subjects consumed the second supplement before HRT. RESULTS: Compared with baseline, REE was elevated significantly in both CHO and PRO at 24 and 48 h after HRT (P < 0.05). At 24 h after HRT, REE in response to PRO was significantly greater compared with CHO (P < 0.05). RER decreased significantly in both CHO and PRO at 24 h after HRT compared with baseline (P < 0.05). No differences were observed in total energy intake, macronutrient intake, or HRT volume (P > 0.05). CONCLUSIONS: Timing PRO before HRT may be a simple and effective strategy to increase energy expenditure by elevating REE the day after HRT. Increasing REE could facilitate reductions in body fat mass and improve body composition if nutritional intake is stable. PMID- 19997004 TI - Effects of vastus medialis oblique retraining versus general quadriceps strengthening on vasti onset. AB - PURPOSE: To compare the effects of vastus medialis oblique (VMO) motor control retraining (MCR) and quadriceps strengthening (QS) exercises on the onset timing of the medial (VMO) and lateral (vastus lateralis, VL) quadriceps muscle. METHODS: This single-blind randomized controlled trial involved 60 currently pain free individuals with a history of anterior knee pain and delayed (>10 ms) onset of VMO relative to VL during stair stepping. A blinded assessor took measures at baseline, immediately after 6 wk of treatment, and after an 8-wk follow-up. Both exercise programs involved weekly individual physiotherapy sessions with home exercises. The MCR program comprised specific VMO exercises incorporating EMG biofeedback, mostly in functional weight-bearing positions. The QS program comprised progressive-resistance inner range open kinetic chain exercises. The primary outcome was the latency between the onset of VMO EMG activity relative to that of VL during stair stepping measured using surface electrodes. RESULTS: During stair ascent, there was a significant change immediately after the intervention in VMO-VL timing in the MCR group only (P = 0.04), but there was no significant difference in the change between groups. During stair descent, VMO-VL timing changed in both groups (P < 0.01), with the MCR group showing a greater change than the QS group (P = 0.02). At the completion of training, quadriceps strength was only improved in the QS group (all P < 0.001). At follow-up, VMO timing and quadriceps strength had improved in both groups compared with baseline (P < 0.01), but there was no difference between groups. CONCLUSIONS: Although greater changes in motor control during stair descent and strength are induced by interventions that target each of these parameters in the short term, both parameters are similarly improved after the cessation of training, regardless of the target of the intervention. PMID- 19997005 TI - Effect of exercise protocol on deoxy[Hb + Mb]: incremental step versus ramp exercise. AB - PURPOSE: The aim of the present study was to investigate whether the sigmoid pattern of deoxy[Hb + Mb] during incremental exercise is specific to non-steady state conditions. METHODS: Ten highly trained cyclists performed an incremental step (40 W x 3 min(-1)) and ramp (35 W x min(-1)) exercise. Deoxy[Hb + Mb] was measured at the distal and proximal sites of the musculus vastus lateralis throughout the exercises using near-infrared spectroscopy. Deoxy[Hb + Mb] was set out as a function of work rate (% peak power), and using curve-fitting techniques, the best-fitting model was determined. RESULTS: These procedures showed that the sigmoid pattern also provided the best fit for the pattern of deoxy[Hb + Mb] in the step exercise. Furthermore, it was observed that the sigmoid model was similar for the ramp (d = 6.9% +/- 1.1% and 6.9% +/- 1.4% x %( 1) peak power; c/d = 52.1% +/- 3.8% and 52.1% +/- 4.5% peak power, for the proximal and distal measurement sites, respectively) and the step exercise (d = 7.4% +/- 1.5% and 6.4% +/- 1.5% x %(-1) peak power; c/d = 52.3% +/- 6.0% and 52.5% +/- 4.2% peak power, for the proximal and distal measurement sites, respectively). The pattern of deoxy[Hb + Mb] was not influenced by measurement site. CONCLUSIONS: From the present study, it can be concluded that the sigmoid pattern of deoxy[Hb + Mb] during incremental exercise is not specific to non steady-state conditions. It was hypothesized that this pattern is an expression of a nonlinear Q x m/V x O2m relationship, related to changes in muscle fiber type recruitment. PMID- 19997006 TI - Fitness and exercise as correlates of sleep complaints: is it all in our minds? AB - PURPOSE: Restoring sleep is associated with psychological well-being. In contrast, poor sleep leads to impaired daily cognitive, emotional, and social functioning. Both commonplace and expert opinion hold that exercise has a favorable impact on preventing poor sleep and improving its quality. However, the scientific basis for this opinion remains limited, and results are mixed. The aim of the present study, therefore, was to explore the impact of perceived physical fitness, exercise, and a perceived lack of activity on sleep in early adulthood. Gender-related patterns were also examined. METHODS: A total of 862 participants (639 females and 223 males; mean +/- SD = 24.67 +/- 5.91 yr) took part in the study. Respondents completed a series of self-report questionnaires assessing perceived physical fitness, exercise, perceived lack of physical activity, insomnia (Insomnia Severity Index), dysfunctional sleep-related thoughts (Fragebogen zur Erfassung allgemeiner Personlichkeitsmerkmale Schlafgestorter), and quality of sleep (Pittsburgh Sleep Quality Index). RESULTS: High perceived physical fitness, but not exercise, was associated with favorable scores for various sleep indicators. A perceived lack of physical activity was associated with poor sleep. Perceived physical fitness and exercise were moderately correlated. Compared with males, females reported more sleep difficulties and also more dysfunctional sleep-related thoughts. CONCLUSIONS: For early adulthood, findings did not support commonplace or expert opinion that exercise behavior has a favorable influence on sleep. Rather, the findings lend support to the importance of cognitive processes in the onset and maintenance of sleep complaints. PMID- 19997007 TI - Potent protective effect conferred by four bouts of low-intensity eccentric exercise. AB - PURPOSE: It is known that submaximal eccentric exercise does not confer as strong a protective effect as maximal eccentric exercise. This study tested the hypothesis that four bouts of submaximal eccentric exercise would confer a similar protective effect to one bout maximal eccentric exercise. METHODS: Thirty untrained men were placed into 4 x 40% (40%) or control (CON) groups (n = 15 per group) by matching preexercise maximal voluntary isometric contraction strength (MVC). The 40% group performed 30 eccentric contractions with a load of 40% MVC (40% ECC) every 2 wk for four times followed 2 wk later by 30 maximal eccentric exercise (100% ECC) of the elbow flexors of the nondominant arm. The CON group performed two bouts of the 100% ECC separated by 2 wk. MVC at six angles, optimum angle (OA), concentric isokinetic strength (30 degrees x s(-1) and 300 degrees x s(-1)), range of motion, upper arm circumference, plasma creatine kinase activity and myoglobin concentration, muscle soreness, and echo intensity of B-mode ultrasound images were taken before to 5 d after each exercise. RESULTS: No significant differences in the changes in any measures were evident between the 100% ECC of the 40% group and the second 100% ECC of the CON group. Changes in all measures except for OA and upper arm circumference after the second to the fourth 40% ECC bouts were significantly smaller than those after the first 40% ECC bout. The changes in the measures after any of the 40% ECC bouts were significantly (P < 0.05) smaller than those after the first 100% ECC bout of the CON group. CONCLUSIONS: These results suggest that repeating submaximal eccentric exercise confers the same magnitude of protective effect as one bout of maximal eccentric exercise against the subsequent maximal eccentric exercise. PMID- 19997008 TI - Abnormal neurovascular control in anabolic androgenic steroids users. AB - PURPOSE: Previous studies showed that anabolic androgenic steroids (AAS) increase vascular resistance and blood pressure (BP) in humans. In this study, we tested the hypotheses 1) that AAS users would have increased muscle sympathetic nerve activity (MSNA) and reduced forearm blood flow (FBF) compared with AAS nonusers and 2) that there would be an association between MSNA and 24-h BP. METHODS: Twelve AAS users aged 31 +/- 2 yr (means +/- SE) and nine age-matched AAS nonusers aged 29 +/- 2 yr participated in the study. All individuals were involved in strength training for at least 2 yr. AAS was determined by urine test (chromatography-mass spectrometry). MSNA was directly measured by microneurography technique. FBF was measured by venous occlusion plethysmography. BP monitoring consisted of measures of BP for 24 h. RESULTS: MSNA was significantly higher in AAS users than that in AAS nonusers (29 +/- 3 vs 20 +/- 1 bursts per minute, P = 0.01). FBF (1.92 +/- 0.17 vs 2.77 +/- 0.24 mL x min(-1) x 100 mL(-1), P = 0.01) and forearm vascular conductance (2.01 +/- 0.17 vs 2.86 +/- 0.31 U, P = 0.02) were significantly lower in AAS users than that in AAS nonusers. Systolic (131 +/- 4 vs 120 +/- 3 mm Hg, P = 0.001), diastolic (74 +/- 4 vs 68 +/- 3 mm Hg, P = 0.02), and mean BP (93 +/- 4 vs 86 +/- 3 mm Hg, P = 0.005) and heart rate (74 +/- 3 vs 68 +/- 3 bpm, P = 0.02) were significantly higher in AAS users when compared with AAS nonusers. Further analysis showed that there was a significant correlation between MSNA and 24-h mean BP (r = 0.75, P = 0.002). CONCLUSIONS: AAS increases MSNA and reduces muscle blood flow in young individuals. In addition, the increase in BP levels in AAS users is associated with augmented sympathetic outflow. These findings suggest that AAS increases the susceptibility for cardiovascular disease in humans. PMID- 19997009 TI - Tackle characteristics and injury in a cross section of rugby union football. AB - BACKGROUND: The tackle is the game event in rugby union most associated with injury. This study's main aims were to measure tackle characteristics from video using a qualitative protocol, to assess whether the characteristics differed by level of play, and to measure the associations between tackle characteristics and injury. METHODS: A cohort study was undertaken. The cohort comprised male rugby players in the following levels: younger than 15 yr, 18 yr, and 20 yr, grade, and elite (Super 12 and Wallabies). All tackle events and technique characteristics were coded in 77 game halves using a standardized qualitative protocol. Game injuries and missed-game injuries were identified and correlated with tackle events. RESULTS: A total of 6618 tackle events, including 81 resulting in a game injury, were observed and coded in the 77 game halves fully analyzed (145 tackle events per hour). An increase in the proportion of active shoulder tackles was observed from younger than 15 yr (13%) to elite (31%). Younger players engaged in more passive tackles and tended to stay on their feet more than experienced players. Younger than 15 yr rugby players had a significantly lower risk of tackle game injury compared with elite players. No specific tackle technique was observed to be associated with a significantly increased risk of game injury. There was a greater risk of game injury associated with two or more tacklers involved in the tackle event, and the greatest risk was associated with simultaneous contact by tacklers, after adjusting for level of play. CONCLUSIONS: Tackle characteristics differed between levels of play. The number of tacklers and the sequence of tackler contact with the ball carrier require consideration from an injury prevention perspective. PMID- 19997010 TI - Test of the classic model for predicting endurance running performance. AB - PURPOSE: To compare the classic physiological variables linked to endurance performance (VO2max, %VO2max at lactate threshold (LT), and running economy (RE)) with peak treadmill velocity (PTV) as predictors of performance in a 16-km time trial. METHODS: Seventeen healthy, well-trained distance runners (10 males and 7 females) underwent laboratory testing to determine maximal oxygen uptake (VO2max), RE, percentage of maximal oxygen uptake at the LT (%VO2max at LT), running velocity at LT, and PTV. Velocity at VO2max (vVO2max) was calculated from RE and VO2max. Three stepwise regression models were used to determine the best predictors (classic vs treadmill performance protocols) for the 16-km running time trial. RESULTS: Simple Pearson correlations of the variables with 16-km performance showed vVO2max to have the highest correlation (r = -0.972) and %VO2max at the LT the lowest (r = 0.136). The correlation coefficients for LT, VO2max, and PTV were very similar in magnitude (r = -0.903 to r = -0.892). When VO2max, %VO2max at LT, RE, and PTV were entered into SPSS stepwise analysis, VO2max explained 81.3% of the total variance, and RE accounted for an additional 10.7%. vVO2max was shown to be the best predictor of the 16-km performance, accounting for 94.4% of the total variance. The measured velocity at VO2max (PTV) was highly correlated with the estimated velocity at vVO2max (r = 0.8867). CONCLUSIONS: Among well-trained subjects heterogeneous in VO2max and running performance, vVO2max is the best predictor of running performance because it integrates both maximal aerobic power and the economy of running. The PTV is linked to the same physiological variables that determine vVO2max. PMID- 19997011 TI - Estimating MET values using the ratio of HR for persons with paraplegia. AB - The current compendium of physical activity (CPA) cannot be applied to persons with disabilities due to the lack of physical activity (PA) they are regularly engaged in and inaccurate MET values when applied to persons with disabilities. PURPOSE: The purposes of this study were (a) to determine whether HR ratio during PA and resting can be used to accurately predict MET values of PA in persons with paraplegia, (b) to compare individual calibration (IC) with group calibration (GC) in error reduction, and (c) to examine prediction generalizability through a cross-validation design. METHODS: Twenty-seven participants (aged 18-45 yr) with complete and incomplete paraplegia at T6 to L4 participated in this study. Oxygen uptake (VO2) and HR were measured simultaneously at rest and during 10 PA using indirect calorimetry and a Polar HR monitor. Predicted METs were calculated using the HR ratio for six activities by applying regression analysis by group (GC) and individuals (IC), respectively. The derived equations were then cross-validated using the four other activities, and corresponding METs were calculated. Absolute error rates (AC), paired t-test, and correlation (r) were used to determine the absolute and relative difference between observed and predicted METs. RESULTS: The overall correlation coefficient (r) between HR ratio and observed METs was 0.77 using group regression and 0.93 +/- 0.05 using individual regression. GC (R2 = 0.59, AC = 0.07%-65.25%) was less accurate than IC (R2 = 0.90 +/- 0.10, AC = 1.64%-10.26%). Cross-validation results also showed higher correlations for IC (r = 0.90 in IC and 0.72 in GC) between observed and predicted METs. CONCLUSIONS: HR ratio was able to accurately predict METs of persons with paraplegia. IC estimated METs more accurately than GC. PMID- 19997012 TI - Reflex inhibition of electrically induced muscle cramps in hypohydrated humans. AB - INTRODUCTION: Anecdotal evidence suggests that ingesting small volumes of pickle juice relieves muscle cramps within 35 s of ingestion. No experimental evidence exists supporting the ingestion of pickle juice as a treatment for skeletal muscle cramps. METHODS: On two different days (1 wk apart), muscle cramps were induced in the flexor hallucis brevis (FHB) of hypohydrated male subjects (approximately 3% body weight loss and plasma osmolality approximately 295 mOsm x kg(-1) H2O) via percutaneous tibial nerve stimulation. Thirty minutes later, a second FHB muscle cramp was induced and was followed immediately by the ingestion of 1 mL x kg(-1) body weight of deionized water or pickle juice (73.9 +/- 2.8 mL). RESULTS: Cramp duration and FHB EMG activity during the cramp were quantified, as well as the change in plasma constituents. Cramp duration (water = 151.9 +/- 12.9 s and pickle juice = 153.2 +/- 23.7 s) and FHB EMG activity (water = 60% +/- 6% and pickle juice = 68% +/- 9% of maximum voluntary isometric contraction EMG activity) were similar during the initial cramp induction without fluid ingestion (P > 0.05). During FHB muscle cramp induction combined with fluid ingestion, FHB EMG activity was again similar (water = 55% +/- 9% and pickle juice = 66% +/- 9% of maximum voluntary isometric contraction EMG activity, P > 0.05). However, cramp duration was 49.1 +/- 14.6 s shorter after pickle juice ingestion than water (84.6 +/- 18.5 vs 133.7 +/- 15.9 s, respectively, P < 0.05). The ingestion of water or pickle juice had little impact on plasma composition 5 min after ingestion. CONCLUSIONS: Pickle juice, and not deionized water, inhibits electrically induced muscle cramps in hypohydrated humans. This effect could not be explained by rapid restoration of body fluids or electrolytes. We suspect that the rapid inhibition of the electrically induced cramps reflects a neurally mediated reflex that originates in the oropharyngeal region and acts to inhibit the firing of alpha motor neurons of the cramping muscle. PMID- 19997013 TI - Physiological effects of tapering and detraining in world-class kayakers. AB - PURPOSE: This study analyzed changes in neuromuscular, body composition, and endurance markers during 4 wk of tapering and subsequent 5 wk of reduced training (RT) or training cessation (TC). METHODS: Fourteen world-class kayakers were randomly assigned to either a TC (n = 7) or an RT group (n = 7). One-repetition maximum (1RM) strength, mean concentric velocity with 45% 1RM (V45%) in the bench press (BP) and prone bench pull (PBP) exercises, and body composition assessments were conducted at the start (T0) and end (T1) of a 43-wk training program, after tapering for the world championships (T2) and after TC or RT (T3). A graded exercise test on a kayak ergometer for determination of maximal oxygen uptake at T0, T1, and T3 was also performed. RESULTS: After tapering, no significant changes were observed in 1RM or V45%. TC resulted in significantly greater declines in 1RM strength (-8.9% and -7.8%, P < 0.05, respectively, for BP and PBP) than those observed for RT (-3.9% and -3.4%). Decreases in V45% in BP and PBP were larger for TC (-12.6% and -10.0%) than for RT (-9.0% and -6.7%). Increases in sum of eight skinfolds were observed after both TC and RT, whereas declines in maximal aerobic power were lower for RT (-5.6%) than for TC (-11.3%). CONCLUSIONS: Short-term TC results in large decreases in maximal strength and especially V45% in highly trained athletes. These results suggest the need of performing a minimal maintenance program to avoid excessive declines in neuromuscular function in cases where a prolonged break from training is required. PMID- 19997014 TI - Physical fitness, adiposity, and metabolic risk factors in young college students. AB - PURPOSE: The purpose of this study was to assess anthropometry, physical fitness, and serum glucose and lipid levels in college students and to determine the relative impact of fitness level compared with body composition on blood chemistries related to metabolic syndrome. METHODS: College students (n = 564; means +/- SD: age = 19.4 +/- 1.1 yr) participating in the Tufts University Longitudinal Health Study between 2000 and 2007 were examined for the relative impact of fitness level compared with body composition on serum glucose, triglycerides, HDL, LDL, and total cholesterol controlling for age and ethnicity. RESULTS: In this college population, 16.2% were overweight or obese, whereas 60% had body fat percentages above desirable levels. Higher percent body fat was associated with increased cholesterol and LDL in both men and women, and with increased triglyceride and decreased HDL in women (P < 0.05). Increased fitness was associated with increased HDL and decreased triglycerides in women and decreased serum glucose in men (P < 0.05). When dichotomized into "fit" and "unfit" groups, students who were fit demonstrated more optimal levels of serum glucose and lipids independent of percent body fat (P < 0.05). CONCLUSIONS: Several metabolic risk factors are already evident in a young college population, and both increased body fatness and decreased physical fitness were associated with metabolic risk. Being physically fit can confer an added benefit to a healthy body composition. It is therefore important to encourage both healthy weights and fitness in late adolescence as each play important and independent roles in biochemical parameters associated with increased chronic disease risk. PMID- 19997015 TI - Massage impairs postexercise muscle blood flow and "lactic acid" removal. AB - PURPOSE: This study tested the hypothesis that one of the ways sports massage aids muscle recovery from exercise is by increasing muscle blood flow to improve "lactic acid" removal. METHODS: Twelve subjects performed 2 min of strenuous isometric handgrip (IHG) exercise at 40% maximum voluntary contraction to elevate forearm muscle lactic acid. Forearm blood flow (FBF; Doppler and Echo ultrasound of the brachial artery) and deep venous forearm blood lactate and H+ concentration ([La-], [H+]) were measured every minute for 10 min post-IHG under three conditions: passive (passive rest), active (rhythmic exercise at 10% maximum voluntary contraction), and massage (effleurage and petrissage). Arterialized [La-] and [H+] from a superficial heated hand vein was measured at baseline. RESULTS: Data are presented as mean +/- SE. Venoarterial [La-] difference ([La-]v-a) at 30 s of post-IHG was the same across conditions (passive = 6.1 +/- 0.6 mmol x L(-1), active = 5.7 +/- 0.6 mmol x L(-1), massage = 5.5 +/- 0.6 mmol x L(-1), NS), whereas FBF was greater in passive (766 +/- 101 mL x min( 1)) versus active (614 +/- 62 mL x min(-1), P = 0.003) versus massage (540 +/- 60 mL x min(-1), P < 0.0001). Total FBF area under the curve (AUC) for 10 min after handgrip was significantly higher in passive versus massage (4203 +/- 531 vs 3178 +/- 304 mL, P = 0.024) but not versus active (3584 +/- 284 mL, P = 0.217). La(-)- efflux (FBF x [La-]v-a) AUC mirrored FBF AUC (passive = 20.5 +/- 2.8 mmol vs massage = 14.7 +/- 1.6 mmol, P = 0.03, vs active = 15.4 +/- 1.9 mmol, P = 0.064). H+ efflux (FBF x [H+]v-a) was greater in passive versus massage at 30 s (2.2 +/- 0.4e(-5) vs 1.3 +/- 0.2e(-5) mmol, P < 0.001) and 1.5 min (1.0 +/- 0.2e(-5) vs 0.6 +/- 0.09e(-5) mmol, P = 0.003) after IHG. CONCLUSIONS: Massage impairs La(-) and H+ removal from muscle after strenuous exercise by mechanically impeding blood flow. PMID- 19997016 TI - Time course of neuromuscular changes during running in well-trained subjects. AB - PURPOSE: Prolonged exercise reduces the capacity of the neuromuscular system to produce force, which is known as fatigue. The purpose of this study was to examine the time course of neural and contractile processes during a 20-km running bout. METHODS: Eight experienced runners (mean T SD: age = 31 T 6 yr, VO2max = 60.1 T 2.2 mL x kg(-1) x min(-1)) completed an all-out self-paced 20-km treadmill run. Isometric knee extensor torque and EMG responses of the vastus lateralis (VL) in response to percutaneous electrical stimulation and voluntary contraction were measured before and after 5, 10, 15, and 20 km of exercise. RESULTS: Participant's RPE, measured using the Borg 6-20 scale, increased steadily throughout the run to a value of 18 T 1 at exercise termination. Maximal voluntary contraction (MVC) of the knee extensors only decreased during the final 5 km of running, with a 15% +/- 12% (P = 0.02) decrease at 20 km. Vastus lateralis EMG during an MVC was reduced after 15 km (-18% +/- 21%, P = or <0.01) and 20 km (-20% +/- 22%, P = 0.03). A significant correlation (r = 0.71, P = 0.048) was observed between the final reduction in MVC and the maximal EMG. Voluntary activation, estimated by the twitch interpolation technique, decreased by 13% +/- 6% at 20 km (P = or < 0.01), and this was significantly correlated (r = 0.70, P = 0.049) with MVC loss. There were no significant changes in the amplitude of the electrically evoked muscle action potential (M-wave) or potentiated twitch during or after the 20-km run. CONCLUSIONS: A reduction in knee extensor MVC only occurs during the final 5 km of a 20-km self-paced run. Impaired voluntary activation and neural drive but not contractile processes are responsible for this decreased strength. PMID- 19997017 TI - Force-velocity relationship in cycling revisited: benefit of two-dimensional pedal forces analysis. AB - PURPOSE: Maximal cycling exercise has been widely used to describe the power velocity characteristics of lower-limb extensor muscles. This study investigated the contribution of each functional sector (i.e., extension, flexion, and transitions sectors) on the total force produced over a complete pedaling cycle. We also examined the ratio of effective force to the total pedal force, termed index of mechanical effectiveness (IE), in explaining differences in power between subjects. METHODS: Two-dimensional pedal forces and crank angles were measured during a cycling force-velocity test performed by 14 active men. Mean values of forces, power output, and IE over four functional angular sectors were assessed: top = 330 degrees -30 degrees , downstroke = 30 degrees -150 degrees , bottom = 150 degrees -210 degrees , and upstroke = 210 degrees -330 degrees . RESULTS: Linear and quadratic force-velocity and power-velocity relationships were obtained for downstroke and upstroke. Maximal power output (Pmax) generated over these two sectors represented, respectively, 73.6% +/- 2.6% and 10.3% +/- 1.8% of Pmax assessed over the entire cycle. In the whole group, Pmax over the complete cycle was significantly related to Pmax during the downstroke and upstroke. IE significantly decreased with pedaling rate, especially in bottom and upstroke. There were significant relationships between power output and IE for top and upstroke when the pedaling rate was below or around the optimal value and in all the sectors at very high cadences. CONCLUSIONS: Although data from force velocity test primarily characterize the muscular function involved in the downstroke phase, they also reflect the flexor muscles' ability to actively pull on the pedal during the upstroke. IE influences the power output in the upstroke phase and near the top dead center, and IE accounts for differences in power between subjects at high pedaling rates. PMID- 19997018 TI - No effect of carbohydrate-protein on cycling performance and indices of recovery. AB - PURPOSE: The aim of this study was to determine whether adding protein to a CHO beverage would improve late-exercise cycle time-trial performance over CHO alone. Furthermore, we examined the effects of coingesting protein with CHO during exercise on postexercise markers of sarcolemmal disruption and the recovery of muscle function. METHODS: In a double-blind, crossover design, 12 trained male cyclists performed 120 min of steady-state (SS) cycling at approximately 55% VO2max followed by a time trial lasting approximately 1 h. At 15-min intervals during SS exercise, participants consumed either a CHO or a CHO + protein (CHO + Pro) beverage (providing 65 g x h(-1) CHO or 65 g x h(-1) CHO plus 19 g x h(-1) protein). Twenty-four hours after the onset of the SS cycle, participants completed a maximum isometric strength test. At rest and 24 h postexercise, a visual analog scale was used to determine lower-limb muscle soreness, and blood samples were obtained for plasma creatine kinase concentration. Dietary control was implemented 24 h before and during the time course of each trial. RESULTS: Average power output sustained during time trial was similar for CHO and CHO + Pro, with no effect of treatment on the time to complete the time trial (60:13 +/ 1:33 and 60:51 +/- 2:40 (min:s) for CHO and CHO + Pro, respectively). Postexercise isometric strength significantly declined for CHO (15% +/- 3%) and CHO + Pro (11% +/- 3%) compared with baseline (486 +/- 28 N). Plasma creatine kinase concentrations, and visual analog scale soreness significantly increased at 24 h postexercise, with no difference between treatments. CONCLUSIONS: The present findings suggest that CHO + Pro coingestion during exercise does not improve late-exercise time-trial performance, ameliorate markers of sarcolemmal disruption, or enhance the recovery of muscle function at 24 h postexercise over CHO alone. PMID- 19997019 TI - Body composition and strength changes in women with milk and resistance exercise. AB - PURPOSE: We aimed to determine whether women consuming fat-free milk versus isoenergetic carbohydrate after resistance exercise would see augmented gains in lean mass and reductions in fat mass similar to what we observed in young men. METHODS: Young women were randomized to drink either fat-free milk (MILK: n = 10; age (mean +/- SD) = 23.2 +/- 2.8 yr; BMI = 26.2 +/- 4.2 kg x m(-2)) or isoenergetic carbohydrate (CON: n = 10; age = 22.4 +/- 2.4 yr; BMI = 25.2 +/- 3.8 kg x m(-2)) immediately after and 1 h after exercise (2 x 500 mL). Subjects exercised 5 d x wk(-1) for 12 wk. Body composition changes were measured by dual energy x-ray absorptiometry, and subjects' strength and fasting blood were measured before and after training. RESULTS: CON gained weight after training (CON: +0.86 +/- 0.4 kg, P < 0.05; MILK: +0.50 +/- 0.4 kg, P = 0.29). Lean mass increased with training in both groups (P < 0.01), with a greater net gain in MILK versus CON (1.9 +/- 0.2 vs 1.1 +/- 0.2 kg, respectively, P < 0.01). Fat mass decreased with training in MILK only (-1.6 +/- 0.4 kg, P < 0.01; CON: -0.3 +/- 0.3 kg, P = 0.41). Isotonic strength increased more in MILK than CON (P < 0.05) for some exercises. Serum 25-hydroxyvitamin D increased in both groups but to a greater extent in MILK than CON (+6.5 +/- 1.1 vs +2.8 +/- 1.3 nM, respectively, P < 0.05), and parathyroid hormone decreased only in MILK (-1.2 +/- 0.2 pM, P < 0.01). CONCLUSIONS: Heavy, whole-body resistance exercise with the consumption of milk versus carbohydrate in the early postexercise period resulted in greater muscle mass accretion, strength gains, fat mass loss, and a possible reduction in bone turnover in women after 12 wk. Our results, similar to those in men, highlight that milk is an effective drink to support favorable body composition changes in women with resistance training. PMID- 19997020 TI - Validity of armband measuring energy expenditure in overweight and obese children. AB - PURPOSE: The purpose of this study was to examine the ability of the SenseWear Pro2 Armband (SWA) to accurately assess energy expenditure in free-living overweight or obese children during a 2-wk period by comparison with energy expenditure measured using the doubly labeled water (DLW) method. A second aim was to examine which software version, Innerview Professional 5.1 or Sensewear Professional 6.0, is the most appropriate for use together with SWA in overweight and obese children. METHODS: A random sample of 22 healthy, overweight, or obese children (11 girls and 11 boys) aged 8-11 yr were recruited from an ongoing intervention study. Energy expenditure in free-living conditions was simultaneously assessed with the SWA and DLW methods during a 14-d period. All data from the SWA were analyzed using InnerView Professional software versions 5.1 (SWA 5.1) and 6.1 (SWA 6.1). RESULTS: An accurate estimation in energy expenditure was obtained when SWA 5.1 was used, showing a nonstatistically significant difference corresponding to 17 (1200) kJ x d(-1) compared with the energy expenditure measured using the DLW method. However, when SWA 6.1 was used, a statistically significant (18%) underestimation of energy expenditure was obtained, corresponding to 1962 (1034) kJ x d(-1) compared with the DLW method. CONCLUSIONS: The SWA together with software version 5.1, but not 6.1, is a valid method for accurately measuring energy expenditure at group level of free-living overweight and obese children. PMID- 19997021 TI - Differences in cardiac parameters among elite rowers and subelite rowers. AB - There is significant individual variability in the cardiac adaptation that occurs in response to exercise training. Factors associated with this variability remain incompletely understood. To date, the relationship between the competition level at which athletes participate and their underlying cardiac parameters has not been explored. PURPOSE: The purpose of this study was to determine whether parameters of cardiac structure and function differ significantly among elite competitive rowers (ER), subelite competitive rowers (SR), and sedentary controls (C). METHODS: Cardiac parameters were assessed in ER (n = 20), SR (n = 20), and C (n = 20) using two-dimensional, tissue Doppler, and speckled-tracking echocardiography. RESULTS: Physiologic cardiac remodeling was present in both ER and SR as evidenced by the significant differences in the majority of structural and functional parameters in both rower groups when compared with C. When compared with SR, ER were found to have greater left ventricular (LV) end diastolic volume (76 +/- 6 vs 71 +/- 8 mL x m(-2), P = 0.02), LV mass (150 +/- 11 vs 134 +/- 16 g x m(-2), P = 0.002), and right ventricular (RV) end-diastolic chamber dimensions (15.6 +/- 0.9 vs 13.9 +/- 1.5 cm(2) x m(-2), P < 0.001). Further, ER demonstrated significantly more enhancement of RV systolic function (peak strain = 36% +/- 7% vs 31% +/- 6%, P = 0.008) and late diastolic relaxation in both the LV (A' = 4.2 +/- 1.3 vs 3.2 +/- 0.9 cm x s(-1), P = 0.01) and the RV (A' = 6.6 +/- 1.4 vs 4.3 +/- 1.3 cm x s(-1), P < 0.001). CONCLUSIONS: Although cardiac remodeling occurs in both ER and SR, specific aspects of cardiac structure and function differ between rowers who compete at the elite and the subelite levels of sport. PMID- 19997022 TI - Physical activity, antioxidant status, and protein modification in adolescent athletes. AB - Exercise may increase reactive oxygen species production, which might impair cell integrity and contractile function of muscle cells. However, little is known about the effect of regular exercise on the antioxidant status of adolescents. PURPOSE: This study aimed to evaluate the impact of exercise on the antioxidant status and protein modifications in adolescent athletes. METHODS: In 90 athletes and 18 controls (16 +/- 2 yr), exercise-related energy expenditure was calculated on the basis of a 7-d activity protocol. Antioxidant intake and plasma concentrations of alpha-tocopherol, carotenoids, and uric acid were analyzed. Plasma antioxidant activity was determined by Trolox equivalent (TE) antioxidant capacity and electron spin resonance spectrometry. Protein modifications were assessed with structural changes of transthyretin using a matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. Data were analyzed by two way ANOVA and post hoc by the Tukey-Kramer test (alpha = 0.05). RESULTS: Antioxidant intake correlated with energy intake and was within the recommended daily allowance for vitamins C and E and beta-carotene. Plasma levels of neither nutritional antioxidants nor uric acid differed between the groups. TE antioxidant capacity was higher in athletes (men = 1.47 +/- 0.2 mmol TE per liter, women = 1.45 +/- 0.2 mmol TE per liter) compared with controls (men = 1.17 +/- 0.04 mmol TE per liter, women = 1.14 +/- 0.04 mmol TE per liter) and increased with exercise-related energy expenditure (P = 0.007). Transthyretin cysteinylation rate differed between the groups, with the highest rate of protein modifications in moderately active subjects (P = 0.007). CONCLUSIONS: Results suggest that if the nutritional choice of athletes is well balanced, enough antioxidants are provided to meet recommended amounts. Moreover, regular exercise increases blood antioxidant capacity in young athletes, whereas chronic exercise was not shown to promote protein modifications. Thus, in young athletes who are sufficiently supplied with antioxidants, beneficial effects of exercise on antioxidant status rather than on oxidative stress may be anticipated. PMID- 19997024 TI - Sedentary behaviors and abnormal glucose tolerance among pregnant Latina women. AB - Studies conducted among nonpregnant populations have observed an association between sedentary behavior and glucose intolerance. Few studies have investigated this association during pregnancy, particularly among Latina women, a population with higher rates of sedentary behavior and abnormal glucose tolerance (AGT) as compared with non-Latina white women. OBJECTIVE: The purpose of this study was to estimate the association between sedentary behavior and AGT. RESEARCH DESIGN AND METHODS: We used data from the Latina gestational diabetes mellitus study, a prospective cohort of 1231 Latina prenatal care patients in western Massachusetts. Sedentary behavior was assessed by bilingual interviewers using a modified version of the Kaiser Physical Activity Survey in prepregnancy, early pregnancy, and midpregnancy and defined as hours spent TV watching, frequency of sitting at work, and participation in sports or exercise activity as well as a derived composite of total sedentary activity. AGT was measured at 24-28 wk of gestation and abstracted from medical records after delivery. Multivariate logistic regression was used to calculate odds ratios and 95% confidence intervals. RESULTS: A total of 12% (n = 119) of participants were classified as having AGT. After adjusting for AGT risk factors, sedentary behaviors in prepregnancy or in early pregnancy were not associated with AGT. However, in midpregnancy, low levels of participation in sports or exercise (odds ratio = 2.01, 95% CI = 1.01-4.02) and increasing total sedentary activity (Ptrend = 0.003) were associated with increased risk for AGT. CONCLUSIONS: Findings suggest an association between sedentary activity in midpregnancy and risk of AGT in this understudied population. PMID- 19997025 TI - Concurrent endurance and strength training not to failure optimizes performance gains. AB - PURPOSE: The purpose of this study was to examine the efficacy of 8 wk of resistance training to failure versus not to failure training regimens at both moderate and low volumes for increasing upper-body strength and power as well as cardiovascular parameters into a combined resistance and endurance periodized training scheme. METHODS: Forty-three trained male rowers were matched and then randomly assigned to four groups that performed the same endurance training but differed on their resistance training regimen: four exercises leading to repetition failure (4RF; n = 14), four exercises not leading to failure (4NRF; n = 15), two exercises not to failure (2NRF; n = 6), and control group (C; n = 8). One-repetition maximum strength and maximal muscle power output during prone bench pull (BP), average power during a 20-min all-out row test (W 20 min), average row power output eliciting a blood lactate concentration of 4 mmol x L( 1) (W 4 mmol x L(-1)), and power output in 10 maximal strokes (W 10 strokes) were assessed before and after 8 wk of periodized training. RESULTS: 4NRF group experienced larger gains in one- repetition maximum strength and muscle power output (4.6% and 6.4%, respectively) in BP compared with both 4RF (2.1% and j1.2%) and 2NRF (0.6% and -0.6%). 4NRF and 2NRF groups experienced larger gains in W 10 strokes (3.6% and 5%) and in W 20 min (7.6% and 9%) compared with those found after 4RF (-0.1% and 4.6%), whereas no significant differences between groups were observed in the magnitude of changes in W 4 mmol x L(-1) (4NRF = 6.2%, 4RF = 5.3%, 2NRF = 6.8%, and C = 4.5%). CONCLUSIONS: An 8-wk linear periodized concurrent strength and endurance training program using a moderate number of repetitions not to failure (4NRF group) provides a favorable environment for achieving greater enhancements in strength, muscle power, and rowing performance when compared with higher training volumes of repetitions to failure in experienced highly trained rowers. PMID- 19997026 TI - Comparison between glucose-6-phosphate dehydrogenase-deficient and normal individuals after eccentric exercise. AB - PURPOSE: Theoretically, glucose-6-phosphate dehydrogenase (G6PD)-deficient compared with nondeficient individuals may be less capable of performing physical activities and/or may be more vulnerable to muscle damage and oxidative stress. Therefore, the purpose of the present study was to examine the effects of a resistance muscle-damaging exercise bout on muscle function and damage, redox status in plasma, and erythrocytes and hemolysis. METHODS: Nine males with established G6PD deficiency and nine males with normal G6PD activity performed an eccentric muscle-damaging exercise protocol. Isometric torque, range of motion, delayed onset muscle soreness, and creatine kinase were measured as indices of muscle function and damage. Reduced glutathione, oxidized glutathione, thiobarbituric acid-reactive substances, protein carbonyls, catalase, uric acid, and total antioxidant capacity were measured as indices of blood redox status. Plasma hemoglobin and bilirubin were measured as indices of hemolysis. All measurements conducted before, immediately after, and 1, 2, 3, 4, and 5 d after exercise. RESULTS: All indices measured confirmed that eccentric exercise induced severe muscle damage, oxidative stress, and hemolysis, peaking at 2 and 3 d postexercise. Lower resting levels of reduced glutathione were detected in the G6PD-deficient group compared with the control group. Nevertheless, both the time course and the magnitude of the changes of the selected muscle performance, redox status (both in plasma and in erythrocytes), and hemolysis indices measured were similar between the two groups. CONCLUSIONS: The present study indicates that G6PD-deficient individuals may participate in high-intensity muscle-damaging activities, without a negative impact on muscle function, blood redox status, and hemolysis. PMID- 19997027 TI - Field evaluation of paralympic athletes in selected sports: implications for training. AB - PURPOSE: The purpose of this study was 1) to describe the acute cardiorespiratory and metabolic responses of Paralympic athletes participating in the following five sports: Nordic sit skiing (NS, n = 5), wheelchair distance racing (WR, n = 6), wheelchair basketball (WB, n = 13), wheelchair fencing (WF, n = 6), and wheelchair tennis (WT, n = 4); and 2) to examine the relationship between field test performance and laboratory measures of aerobic fitness of these Paralympic athletes. METHODS: Each athlete completed an incremental arm cranking exercise test to determine ventilatory threshold (VT) and peak oxygen uptake (VO2peak). Subsequently, field assessments were carried out using a telemetric system to measure the cardiorespiratory responses in their respective sport. RESULTS: VT and VO2peak (both expressed in milliliters per kilogram per minute) of athletes competing in NS (38.3 +/- 5.76 and 51.0 +/- 6.92 mL x kg(-1) x min(-1)) and WR (35.5 +/- 5.96 and 48.1 +/- 6.35 mL x kg(-1) x min(-1)) were significantly higher (P < 0.05) than those competing in WB (26.0 +/- 2.13 and 36.9 +/- 3.70 mL x kg( 1) x min(-1)), WF (23.2 +/- 3.96 and 34.4 +/- 5.81 mL x kg(-1) x min(-1)), and WT (24.0 +/- 2.30 and 33.1 +/- 2.85 mL x kg(-1) x min(-1)). In the field tests, the average V O2, higher in NS and WR than in WB, WF, and WT, during NS, WR, WB, WF, and WT was 79.4% +/- 3.30%, 84.4% +/- 2.10%, 72.1% +/- 5.72%, 73.0% +/- 3.10%, and 73.0% +/- 1.91%, respectively, of VO2peak. There was a strong linear relationship between VO2 measured during the field tests and VT and VO2peak (R2 = 0.92 in each case). CONCLUSIONS: Athletes regulated their average work intensity during the field tests in the five Paralympic sports to approximate their individualized VT measured during incremental arm cranking exercise test, and this intensity was within the range recommended by the American College of Sports Medicine to improve cardiorespiratory fitness in well-trained subjects. In addition, performance of Paralympic athletes in these sports was highly dependent upon athletes' aerobic fitness. PMID- 19997028 TI - Loading of trained inspiratory muscles speeds lactate recovery kinetics. AB - PURPOSE: The purpose of this study was to investigate the effects of inspiratory threshold loading (ITL) and inspiratory muscle training (IMT) on blood lactate concentration ([lac(-)]B) and acid-base balance after maximal incremental cycling. METHODS: Eighteen subjects were divided into a control (n = 9) or an IMT group (n = 9). Before and after a 6-wk intervention, subjects completed two maximal incremental cycling tests followed by 20 min of recovery with (ITL) or without (passive recovery (PR)) a constant inspiratory resistance (15 cm H2O). The IMT group performed 6 wk of pressure threshold IMT at 50% maximal inspiratory mouth pressure. Throughout recovery, acid-base balance was quantified using the physicochemical approach by measuring the strong ion difference ([SID] = [Na+] + [K+] - [Cl-] + [lac-]), the total concentration of weak acids ([Atot-]), and the partial pressure of carbon dioxide (PCO2). RESULTS: After the intervention, maximal inspiratory mouth pressure increased in the IMT group only (+34%). No differences in lactate clearance were observed between PR and ITL before the intervention in both groups and after the intervention in the control group. After IMT, relative to PR, [lac-]B was reduced throughout ITL (minutes 2-20) by 0.66 +/- 1.28 mmol x L(-1) (P < 0.05), and both the fast (lactate exchange) and the slow (lactate clearance) velocity constants of the lactate recovery kinetics were increased (P < 0.05). Relative to pre-IMT, ITL reduced plasma [H], which was accounted for by an IMT-mediated increase in [SID] due almost exclusively to a 1.7-mmol x L(-1) reduction in [lac-]B. CONCLUSIONS: After maximal exercise, ITL affected lactate recovery kinetics only after IMT. Our data support the notion that the inspiratory muscles are capable of lactate clearance that increases [SID] and reduces [H+]. These effects may facilitate subsequent bouts of high intensity exercise. PMID- 19997029 TI - Early physical activity provides sustained bone health benefits later in childhood. AB - PURPOSE: This study examined the potential effect of early childhood moderate and vigorous physical activity (MVPA) on later bone health. METHODS: Three hundred and thirty-three children, participating in the Iowa Bone Development Study, were studied at ages 5, 8, and 11 yr. MVPA (min x d(-1)) was measured using an accelerometry-based physical activity monitor. Bone mineral content (BMC; g) of the whole body, lumbar spine, and hip was measured using dual-energy x-ray absorptiometry. Mixed regression models were used to test whether MVPA at age 5 yr had an effect on BMC at ages 8 and 11 yr after adjustment for concurrent height, weight, age, maturity, and MVPA. The analysis was repeated to control for bone outcomes at age 5 yr. Mixed-model least-squares mean values at the person level of covariates for age group were used to compare the BMC at ages 8 and 11 yr of children in the highest and lowest quartiles of MVPA at age 5 yr. RESULTS: For boys and girls, MVPA at age 5 yr predicted BMC adjusted for concurrent height, weight, age, maturity, and MVPA at ages 8 and 11 yr (P < 0.05). When the analysis was repeated to also control for BMC at age 5 yr, the effect of MVPA at age 5 yr was significant for boys but not for girls. Boys and girls in the highest quartile of MVPA at age 5 yr had 4%-14% more BMC at ages 8 and 11 yr than those in the lowest quartile of MVPA at age 5 yr (P < 0.05). CONCLUSIONS: These results provide support for the benefits of early MVPA on sustained bone health during childhood especially for boys. Results indicate the importance of increasing MVPA as a strategy to improve BMC later in childhood. PMID- 19997030 TI - Measuring total and domain-specific sitting: a study of reliability and validity. AB - PURPOSE: Although independent relationships between sitting behaviors (mainly television viewing) and health outcomes have been reported, few studies have examined the measurement properties of self-report sitting questions. This study assessed gender-specific test-retest reliability and validity of a questionnaire that assessed time spent sitting on weekdays and weekend days: 1) traveling to and from places, 2) at work, 3) watching television, 4) using a computer at home, and 5) for leisure, not including television. METHODS: Test-retest reliability of domain-specific sitting time (min x d(-1)) on weekdays and weekend days was assessed using data collected on two occasions (median = 11 d apart). Validity of domain-specific self-reported sitting time on weekdays and weekend days was assessed against log data and sedentary accelerometer data. RESULTS: Complete repeat questionnaire and log data were obtained from 157 women (aged 51-59 yr) and 96 men (aged 45-63 yr). Reliability coefficients were high for weekday sitting time at work, watching television, and using a computer at home (r = 0.84 0.78) but lower for weekend days across all domains (r = 0.23-0.74). Validity coefficients were highest for weekday sitting time at work and using a computer at home (r = 0.69-0.74). With the exception of computer use and watching television for women, validity of the weekend-day sitting time items was low. CONCLUSIONS: This study confirms the importance of measuring domain- and day specific sitting time. The measurement properties of questions that assess structured domain-specific and weekday sitting time were acceptable and may be used in future studies that aim to elucidate associations between domain-specific sitting and health outcomes. PMID- 19997031 TI - Leisure time and occupational physical activity among racial or ethnic minorities. AB - PURPOSE: One of the factors distinguishing non-Latino blacks (NLB) and Latinos from non-Latino whites (NLW) is higher rates of occupational physical activity (OPA) and less participation in leisure time physical activity (LTPA). We examined participation in OPA and LTPA among employed individuals and the relationship between OPA and LTPA across select racial or ethnic groups and Latino subgroups. METHODS: We pooled data from 2000 to 2003 of the National Health Interview Survey. We divided the survey participants into three groups: 1) those with no LTPA, 2) those who reported some LTPA but not for sufficient time and intensity to meet recommended guidelines, and 3) those who reported LTPA at levels that met or exceeded recommendations. We used ordinal logistic regression to examine whether NLB and Latinos or Latino subgroups were less likely to report LTPA than NLW while controlling for social, economic, and demographic factors that may have accounted for group differences. We further examined the prevalence of OPA and the relationship between LTPA and OPA. RESULTS: Among employed individuals, NLB and Latinos had significantly more individuals reporting no LTPA compared with NLW. Latinos had the greatest proportion of individuals reporting no LTPA. Furthermore, it was found that significantly more Latinos had physically active occupations compared with NLB and NLB compared with NLW, respectively. Among employed Latinos, Cubans and Dominicans were most likely to report no LTPA, and Mexicans had the greatest percentage of workers with a physically active occupation. LTPA was not significantly associated with having a physically active occupation across races and Latino subgroups. CONCLUSIONS: Participation in LTPA among ethnic or racial minorities is lower than that of NLW, and the OPA rates are higher. OPA does not significantly impact participation in LTPA in employed adults. PMID- 19997032 TI - Assessment of extravascular lung water and cardiac function in trimix SCUBA diving. AB - An increasing number of recreational self-contained underwater breathing apparatus (SCUBA) divers use trimix of oxygen, helium, and nitrogen for dives deeper than 60 m of sea water. Although it was seldom linked to the development of pulmonary edema, whether SCUBA diving affects the extravascular lung water (EVLW) accumulation is largely unexplored. METHODS: Seven divers performed six dives on consecutive days using compressed gas mixture of oxygen, helium, and nitrogen (trimix), with diving depths ranging from 55 to 80 m. The echocardiographic parameters (bubble grade, lung comets, mean pulmonary arterial pressure (PAP), and left ventricular function) and the blood levels of the N terminal part of pro-brain natriuretic peptide (NT-proBNP) were assessed before and after each dive. RESULTS: Venous gas bubbling was detected after each dive with mean probability of decompression sickness ranging from 1.77% to 3.12%. After each dive, several ultrasonographically detected lung comets rose significantly, which was paralleled by increased pulmonary artery pressure (PAP) and decreased left ventricular contractility (reduced ejection fraction at higher end-systolic and end-diastolic volumes) as well as the elevated NT-proBNP. The number of ultrasound lung comets and mean PAP did not return to baseline values after each dive. CONCLUSIONS: This is the first report that asymptomatic SCUBA dives are associated with accumulation of EVLW with concomitant increase in PAP, diminished left ventricular contractility, and increased release of NT-proBNP, suggesting a significant cardiopulmonary strain. EVLW and PAP did not return to baseline during repetitive dives, indicating possible cumulative effect with increasing the risk for pulmonary edema. PMID- 19997033 TI - Behavioral compensatory adjustments to exercise training in overweight women. PMID- 19997034 TI - METs in adults while playing active video games: a metabolic chamber study. AB - PURPOSE: Active video game systems controlled through arm gestures and motions (Nintendo Wii Sports) and video games controlled through force plate (Wii Fit Plus) are becoming increasingly popular. This study was performed to determine the energy expenditure (EE) during Wii Fit Plus and Wii Sports game activities. METHODS: Twelve adult men and women performed all the activities of Wii Sports (five activities: golf, bowling, tennis, baseball, and boxing) and Wii Fit Plus (63 activities classified as yoga, resistance, balance, and aerobic exercises). Each activity was continued for at least 8 min to obtain a steady-state EE. Because EE was assessed in an open-circuit indirect metabolic chamber consisting of an airtight room (20,000 or 15,000 L), subjects were freed of apparatus to collect expired gas while playing the games. MET value was calculated from resting EE and steady-state EE during activity. RESULTS: The mean MET values of all 68 activities were distributed over a wide range from 1.3 METs (Lotus Focus) to 5.6 METs (single-arm stand). The mean MET values in yoga, balance, resistance, and aerobic exercise of Wii Fit Plus and Wii Sports were 2.1, 2.0, 3.2, 3.4, and 3.0 METs, respectively. Forty-six activities (67%) were classified as light intensity (<3 METs), and 22 activities (33%) were classified as moderate intensity (3.0-6.0 METs). There were no vigorous-intensity activities (>6.0 METs). CONCLUSIONS: Time spent playing one-third of the activities supplied by motion- and gesture-controlled video games can count toward the daily amount of exercise required according to the guidelines provided by the American College of Sports Medicine and the American Heart Association, which focus on 30 min of moderate-intensity daily physical activity 5 d x wk(-1). PMID- 19997035 TI - Exercise during hematopoietic stem cell transplant hospitalization in children. AB - PURPOSE: The purpose of this controlled trial was to assess the effect of an approximately 3-wk intrahospital exercise intervention performed during inpatient hospitalization for pediatric allogeneic hematopoietic stem cell transplant (HSCT) on (i) immune cell recovery and (ii) body composition. METHODS: Immune (i.e., blood counts of leukocytes, monocytes, lymphocytes, and lymphocyte subpopulations) and anthropometric variables (i.e., body mass, body mass index, and estimated fat-free mass) were measured before and after (+15 and 30 d) HSCT. Seven children (5 boys and 2 girls; age (mean +/- SD) = 8 +/- 4 yr) with high risk cancer performed an individualized training program (aerobic + resistance exercises) in their isolated hospital rooms. We also assessed a control group (n = 13; 9 boys and 4 girls; age = 7 +/- 3 yr) with similar medical conditions and following the same transplant protocol. RESULTS: In both groups, the dendritic cell count decreased from pre-HSCT to +15 d post-HSCT and thereafter (up to +30 d) remained stabile; however, the posttransplant decrease was more abrupt in the control group than that in the intervention group (-87% vs -63%, respectively, from pre-HSCT to +15 d). The rest of the immune cell parameters measured showed a similar response from pre-HSCT to post-HSCT in both groups. We found a significant effect of the interaction group x time for all anthropometric variables (weight, body mass index, body fat, and fat-free mass), indicating an increase over the hospitalization period only in the intervention group, for example, body mass increased from 32.9 +/- 18.7 kg pre-HSCT to 35.4 +/- 18.6 kg at +30 d in the intervention group versus a decrease from 30.2 +/- 16.6 to 29.3 +/- 6.3 kg in the control group. CONCLUSION: Our findings support the feasibility of exercise training interventions during hospitalization, including immunocompromised children. PMID- 19997036 TI - Semantic judgment of Chinese-Japanese bilinguals: a near-infrared spectroscopy study. AB - We used functional near-infrared spectroscopy to investigate the role of the dorsolateral prefrontal cortex of bilinguals in semantic processing. Chinese Japanese bilinguals and Japanese monolinguals judged if visually presented pairings of a word and a definition were Japanese. The four types of pairs appeared in Japanese and/or Chinese dictionaries: both, Japanese-specific, Chinese-specific, and neither. The bilinguals showed greater oxy-hemoglobin concentrations in their left dorsolateral prefrontal cortex for the Chinese specific pair, while those in its right homologue were greater for the Japanese specific, the Chinese-specific, and the neither pairs. These results imply that in the bilinguals, the left dorsolateral prefrontal cortex is related to reducing activation of semantic information in the nontarget language, while its right homologue focuses attention on the target language. PMID- 19997037 TI - PC3 is involved in the shift from proliferation to differentiation and maturation in spiral ganglion neurons. AB - PC3 is a member of the BTG/Tob family of antiproliferative genes. Here, we report the results of an analysis of PC3 protein expression in spiral ganglion neurons of the rat cochlea at embryonic days 16 (E16) and 20 (E20), and postnatal days 4 (P4) and 7 (P7). PC3 expression was observed in the cytoplasm of ganglion neurons at E16 and E20, and this protein had translocated to the nucleus by P4. The expression of Ki-67, a nuclear antigen expressed by dividing cells, was detected in ganglion neurons at E16 and E20, but not at P4 or P7. These results suggest that PC3 is involved in the shift from proliferation to differentiation and maturation in the ganglion neurons of the rat cochlea. PMID- 19997038 TI - Ipsilateral primary sensorimotor cortical response to mechanical tactile stimuli. AB - We studied somatosensory-evoked fields elicited by mechanical versus electrical stimuli to index finger of healthy participants. Mechanical stimulation was index pulp compression and decompression by using nonmagnetic mechanical stimulator. Electrical stimulation was three times of sensory threshold and delivered to index pulp by using ball-shaped electrodes. Mechanical/electrical stimuli evoked contralateral primary somatosensory cortical responses in all respective participants. Compressive stimuli evoked ipsilateral primary sensorimotor cortical responses in all respective participants, with dipole strengths less than contralateral primary somatosensory cortical responses of compressive stimuli. Mechanical/electrical stimuli evoked secondary somatosensory (SII) cortical responses bilaterally; electrical-stimuli SII dipole strengths were relatively stronger than compressive-stimuli SII responses. It is concluded that the use of mechanical stimulation may improve our understanding of functional sensory cortical responses compared with electrical stimulation. PMID- 19997039 TI - Parietal cortex and information granularity in labile and stable learning. AB - We investigated the effects of rule learning based on information granularity. Using two homogeneous Boolean arithmetic tasks, we examined parietal cortex activity during the calculation of labile and stabilized learning. The results revealed stability-related behavioral advantages in a comparison of granularity based effects with labile learning of Boolean problems. The functional MRI results revealed that different regions within the parietal cortex exhibited increased activity while solving Boolean problems in both the conditions. The calculation of labile rule learning based on low-granularity Boolean rules was significantly correlated with activation in bilateral parietal cortex, whereas stable rule learning based on high-granularity Boolean rules was correlated with activation in the left parietal cortex. PMID- 19997040 TI - Radiolabeling RGD peptide and preliminary biodistribution evaluation in mice bearing S180 tumors. AB - OBJECTIVE: To prepare the rhenium-188 (188Re)-arginine-glycine-aspartic acid (RGD) peptide in a convenient manner and to evaluate its potential as an agent for alphavbeta3 integrin receptor-positive tumors. METHODS: Radiolabeled RGD was obtained by conjugating the His group at the end of peptide with fac [188Re(H2O)3(CO)3]+. Chelating efficiency of fac-[188Re(H2O)3(CO)3]+ and radiolabeling efficiency of radiolabeled peptide were measured by thin-layer chromatography and high-performance liquid chromatography. In-vitro stability of the radio-complex was determined in phosphate-buffered saline (0.05 mol/l, pH 7.4), new-born calf serum, His or Cys solution at 37 degrees C or room temperature and analyzed by thin-layer chromatography. A biodistribution study was carried out in mice bearing S180 tumors. RESULTS: 188Re-RGD was obtained with a more than 95% of radiolabeling efficiency, and showed high stability in phosphate-buffered saline, new-born calf serum, His and Cys solution. Furthermore, this radio-complex was cleared rapidly from the blood and showed specific tumor uptake in mice bearing S180 tumors. CONCLUSION: 188Re-RGD was prepared by a simple method. Preliminary biodistribution results showed its potential as an agent for cancer therapy and encouraged further investigation. PMID- 19997041 TI - The effect of furosemide dose timing on bladder activity in oncology imaging with 18F-fluorodeoxyglucose PET/CT. AB - OBJECTIVE: To compare the effects of two furosemide administration protocols on bladder activity during 18F-fluorodeoxyglucose (18F-FDG) PET/computed tomography (CT) imaging. METHODS: A total of 109 consecutive patients with known or suspected malignancy, meeting our inclusion criteria, were chosen over a discrete time period. Group 1 (n=39) received furosemide 20 mg intravenous 15 min before PET/CT imaging (i.e. approximately 45 min after 18F-FDG administration). Group 2 (n=45) received furosemide 20 mg intravenous 15 min after 18F-FDG. Group 3 (n=25) did not receive furosemide and served as controls. Bladder standard uptake values (SUVs) and volume, and liver SUV data were collected. RESULTS: Relative to the control group, both furosemide groups showed significantly lower mean and maximum SUV bladder activities (P<0.001), lower mean bladder-to-liver SUV ratios (P<0.001), larger mean bladder volumes (P<0.001) and higher proportions of bladder PET/CT image mis-registration. Patients tolerated earlier administration of furosemide (group 2) better relative to urinary urgency during imaging. CONCLUSION: The use of a relatively simple diuretic protocol can significantly lower bladder FDG activity and potentially improve image quality by reducing bladder activity artifacts and avoid invasive bladder catheterization. Administering furosemide earlier after FDG injection (i.e. 15 min) versus later (i.e. 15 min before imaging) appears to be better tolerated by patients. PMID- 19997042 TI - Cytochrome b5 and NADH cytochrome b5 reductase: genotype-phenotype correlations for hydroxylamine reduction. AB - OBJECTIVES: NADH cytochrome b5 reductase (b5R) and cytochrome b5 (b5) catalyze the reduction of sulfamethoxazole hydroxylamine (SMX-HA), which can contribute to sulfonamide hypersensitivity, to the parent drug sulfamethoxazole. Variability in hydroxylamine reduction could thus play a role in adverse drug reactions. The aim of this study was to characterize variability in SMX-HA reduction in 111 human livers, and investigate its association with single nucleotide polymorphisms (SNPs) in b5 and b5R cDNA. METHODS: Liver microsomes were assayed for SMX-HA reduction activity, and b5 and b5R expression was semiquantified by immunoblotting. The coding regions of the b5 (CYB5A) and b5R (CYB5R3) genes were resequenced. RESULTS: Hepatic SMX-HA reduction displayed a 19-fold range of individual variability (0.06-1.11 nmol/min/mg protein), and a 17-fold range in efficiency (Vmax/Km) among outliers. SMX-HA reduction was positively correlated with b5 and b5R protein content (P<0.0001, r=0.42; P=0.01, r=0.23, respectively), and expression of both proteins correlated with one another (P<0.0001; r=0.74). A novel cSNP in CYB5A (S5A) was associated with very low activity and protein expression. Two novel CYB5R3 SNPs, R59H and R297H, displayed atypical SMX-HA reduction kinetics and decreased SMX-HA reduction efficiency. CONCLUSION: These studies indicate that although novel cSNPs in CYB5A and CYB5R3 are associated with significantly altered protein expression and/or hydroxylamine reduction activities, these low-frequency cSNPs seem to only minimally impact overall observed phenotypic variability. Work is underway to characterize polymorphisms in other regions of these genes to further account for individual variability in hydroxylamine reduction. PMID- 19997043 TI - Variation in the catechol-O-methyltransferase Val 158 Met polymorphism associated with conduct disorder and ADHD symptoms, among adolescent male delinquents. AB - OBJECTIVE: Variation in the catechol-O-methyltransferase gene (COMT) has been associated with antisocial behavior in populations with attention deficit/hyperactivity disorder (ADHD). This study examined whether COMT would predict antisocial behavior in a sample with high levels of behavior problems, not necessarily ADHD. In addition, because previous research suggests that COMT may be associated with ADHD in males, association between COMT and ADHD symptoms was examined. METHOD: This study tested whether variation in three polymorphisms of the COMT gene was predictive of symptoms of conduct disorder and ADHD, in a sample of 174 incarcerated Russian adolescent male delinquents. RESULTS: The Val allele of the ValMet polymorphism was significantly associated with conduct disorder diagnosis and symptoms, whereas the Met allele was associated with ADHD symptoms. CONCLUSION: The ValMet polymorphism of the COMT gene shows a complex relation to behavior problems, influencing conduct disorder and ADHD symptoms in opposite directions in a high-risk population. PMID- 19997044 TI - No association between the serotonin-1A receptor gene single nucleotide polymorphism rs6295C/G and symptoms of anxiety or depression, and no interaction between the polymorphism and environmental stressors of childhood anxiety or recent stressful life events on anxiety or depression. AB - BACKGROUND: There are conflicting reports of an association between a common functional single nucleotide polymorphism (rs6295C/G) in the serotonin-1A receptor gene (HTR1A) and psychological disorders. METHODS: In our study we investigated associations between this polymorphism and symptoms of anxiety and depression in a population sample of 6445 Caucasians in the age groups 20-24, 40 44, and 60-64 years. We also looked for interactions between the polymorphism and environmental stressors of childhood adversity or recent stressful life events on anxiety and depression. RESULTS: There were no significant associations between the polymorphism and anxiety, depression or associated personality traits in the three age cohorts. There were no significant gene-environment interactions between the polymorphism and either of the environmental stressors on anxiety or depression. CONCLUSION: We found no associations or gene-environment interactions involving the polymorphism and symptoms of anxiety or depression. PMID- 19997045 TI - M1 protein from streptococcus pyogenes induces nitric oxide-mediated vascular hyporesponsiveness to phenylephrine: involvement of toll-like receptor activation. AB - Streptococcus pyogenes carrying M1 protein causes the severe and increasingly prevalent streptococcal toxic shock syndrome and necrotizing fasciitis. M1 protein is an important virulence factor of S. pyogenes and induces an inflammatory response in human monocytes. We wanted to investigate if purified M1 protein in solution could induce vascular NO production leading to vasopressor hyporesponsiveness. Rat aortic segments were incubated with M1 protein or LPS in vitro. M1 protein (10 microg mL) and LPS (1 ng mL) to a similar extent induced NO production and hyporesponsiveness to the vasoconstrictor phenylephrine. Immunogold electron microscopy demonstrated that M1 protein binds to Toll-like receptor 2 (TLR2) as well as TLR4 in mouse aorta but only to TLR2 in human omental artery. Incubation with M1 protein caused a reduction in the contractile response to phenylephrine in aortic segments from wild-type and TLR2-knockout but not from TLR4-knockout mice. In conclusion, M1 protein causes vascular NO production leading to hyporesponsiveness to vasopressors via a mechanism involving TLR, but the subtypes may be species dependent. M1 protein could contribute to the circulatory disturbances accompanying severe invasive streptococcal infections. PMID- 19997046 TI - Protective effects of hydrogen gas on murine polymicrobial sepsis via reducing oxidative stress and HMGB1 release. AB - Despite recent advances in antibiotic therapy and intensive care, sepsis is still considered to be the most common cause of death in intensive care units. Excessive production of reactive oxygen species plays an important role in the pathogenesis of sepsis. Recently, it has been suggested that molecular hydrogen (H2) exerts a therapeutic antioxidant activity by selectively reducing hydroxyl radicals (*OH, the most cytotoxic reactive oxygen species) and effectively protects against organ damage induced by I/R. Therefore, we hypothesized that H2 treatment had a beneficial effect on sepsis. In the present study, we found that H2 inhalation starting at 1 and 6 h after cecal ligation and puncture (CLP) or sham operation significantly improved the survival rate of septic mice with moderate or severe CLP in a concentration- and time-dependent manner. Furthermore, moderate or severe CLP mice showed significant multiple organ damage characterized by the increases of lung myeloperoxidase activity, wet-to-dry weight ratio, protein concentration in bronchoalveolar lavage, serum biochemical parameters, and organ histopathologic scores at 24 h after CLP operation, which was significantly attenuated by 2% H2 treatment. In addition, we found that the beneficial effects of H2 treatment on sepsis and sepsis-associated organ damage were associated with the decreased levels of oxidative product, increased activities of antioxidant enzymes, and reduced levels of high-mobility group box 1 in serum and tissue. Thus, H2 inhalation may be an effective therapeutic strategy for patients with sepsis. PMID- 19997047 TI - Immunomodulatory properties of pentoxifylline are mediated via adenosine dependent pathways. AB - The phosphodiesterase inhibitor pentoxifylline (PTX) exerts multiple beneficial immunomodulatory effects in states of hyperinflammation. However, the exact mechanism of action still remains elusive, and the clinical effects of PTX cannot be reliably predicted. In immune cells, the G protein-coupled adenosine A2A receptor (A2AR) exerts strong anti-inflammatory effects. As PTX amplifies signaling pathways downstream of Gs protein-coupled receptors, the A2AR-signaling pathway might be involved in the mediation of immune-suppressive effects of PTX. Here, we investigated this assumption in LPS-stimulated human polymorphonuclear (PMN) leukocytes and in anti-CD3/CD28-stimulated human T cells. In stimulated PMN leukocytes, PTX treatment led to a 4.5-fold decrease of the 50% inhibitory concentrations of adenosine on the H2O2 production; i.e., for adenosine plus PTX (in clinically relevant concentrations), an overadditive increase of inhibitory effects from less than 20% (estimated for each) to 56% (+/-5%) was found. In T cells, adenosine plus PTX revealed similar synergistic inhibitory effects on proinflammatory cytokine production. Inhibition of interferon gamma and TNF-alpha production increased from 7% (+/-1%) and 31% (+/-6%) (PTX alone) to 49% (+/-2%) and 69% (+/-6%), respectively. In T cells and PMN leukocytes, mRNA transcription of the A2AR was significantly increased upon stimulation, which was not influenced by PTX. In human PMN leukocytes and T cells, clinically relevant anti inflammatory effects of PTX can be achieved only in the presence of sufficient adenosine concentrations. Sufficient adenosine levels might be a prerequisite for the accessibility of sepsis patients to treatment with PTX. PMID- 19997048 TI - 15-deoxy-Delta12,14 prostaglandin GJ2 but not rosiglitazone regulates metalloproteinase 9, NOS-2, and cyclooxygenase 2 expression and functions by peroxisome proliferator-activated receptor gamma-dependent and -independent mechanisms in cardiac cells. AB - Sepsis or endotoxemia produced by LPS followed by hypotension and multiorganic failure may lead to cardiac dysfunction contributing to mortality. Cardiac failure is usually associated to activation of nuclear factor kappaB (NF-kappaB) and mitogen-activated protein kinase (MAPK), which play an important role in proinflammatory enzymes expression. It has been shown that 15-deoxy-Delta12,14 prostaglandin J2 (15dPGJ2) can repress the inflammatory response by means of peroxisome proliferator-activated receptor gamma (PPARgamma)-dependent and independent mechanisms. However, its precise role in heart is poorly understood. In the present study, mouse neonatal cardiomyocytes were isolated and stimulated with LPS to investigate the role of PPARgamma-specific ligands 15dPGJ2 and rosiglitazone on cardiac inflammatory response. Inducible NO synthase, cyclooxygenase 2, and metalloproteinase 9 mRNA levels, protein expression, and activity were inhibited with 15dPGJ2 but not by rosiglitazone. Peroxisome proliferator-activated receptor gamma antagonist, GW9662, prevented all these 15dPGJ2 actions. To go inside the mechanisms by which 15dPGJ2 exerts inhibitory effects, cells were preincubated with specific chemical inhibitors of NF-kappaB and p38 MAPK, and we found that these signaling cascades are implicated in 15dPGJ2 action as well as PPARgamma. These results suggest that only the natural PPARgamma ligand, 15dPGJ2, but not the synthetic one, rosiglitazone, regulates the inflammatory response by inhibition of inducible NO synthase, cyclooxygenase 2, and metalloproteinase 9 expression. Moreover, our results offer an additional 15dPGJ2 mechanism of action, despite PPARgamma, showing NF-kappaB and p38 MAPK participation. PMID- 19997049 TI - Nos3 protects against systemic inflammation and myocardial dysfunction in murine polymicrobial sepsis. AB - NO has been implicated in the pathogenesis of septic shock. However, the role of NO synthase 3 (NOS3) during sepsis remains incompletely understood. Here, we examined the impact of NOS3 deficiency on systemic inflammation and myocardial dysfunction during peritonitis-induced polymicrobial sepsis. Severe polymicrobial sepsis was induced by colon ascendens stent peritonitis (CASP) in wild-type (WT) and NOS3-deficient (NOS3KO) mice. NOS3KO mice exhibited shorter survival time than did WT mice after CASP. NOS3 deficiency worsened systemic inflammation assessed by the expression of inflammatory cytokines in the lung, liver, and heart. Colon ascendens stent peritonitis markedly increased the number of leukocyte infiltrating the liver and heart in NOS3KO but not in WT mice. The exaggerated systemic inflammation in septic NOS3KO mice was associated with more marked myocardial dysfunction than in WT mice 22 h after CASP. The detrimental effects of NOS3 deficiency on myocardial function after CASP seem to be caused by impaired Ca handling of cardiomyocytes. The impaired Ca handling of cardiomyocytes isolated from NOS3KO mice subjected to CASP was associated with depressed mitochondrial ATP production, a determinant of the Ca cycling capacity of sarcoplasmic reticulum Ca-ATPase. The NOS3 deficiency-induced impairment of the ability of mitochondria to produce ATP after CASP was at least in part attributable to reduction in mitochondrial respiratory chain complex I activity. These observations suggest that NOS3 protects against systemic inflammation and myocardial dysfunction after peritonitis-induced polymicrobial sepsis in mice. PMID- 19997050 TI - Simvastatin reduces mortality and hepatic injury after hemorrhage/resuscitation in rats. AB - Statins are established in the prevention and therapy of chronic cardiovascular diseases because of inhibition of HMG-CoA (3-hydroxy-3-methylglutaryl-coenzyme A), thus lowering blood cholesterol levels. However, their cholesterol independent effects include regulation of Rho/Rho-kinases (ROCK) and eNOS, proteins centrally involved in various models of acute inflammation. Therefore, we tested the hypothesis that simvastatin confers protection after rat hemorrhage/resuscitation (H/R) and wanted to elucidate the mechanisms involved. Fifty-two female Lewis rats (180-250 g) were pretreated with simvastatin 5 mg/kg per day or vehicle for 6 days (i.p.). Then, rats were hemorrhaged to a mean arterial pressure of 30 +/- 2 mmHg for 60 min and resuscitated. Control group underwent surgical procedures without H/R. Two hours after resuscitation, tissues were harvested. Mortality was assessed 72 h after H/R. Simvastatin pretreatment increased survival after H/R from 20% to 80%. Serum alanine aminotransferase after H/R increased 2.2-fold in vehicle as compared with simvastatin-treated rats. Histopathological analysis revealed decreased hepatic necrosis in simvastatin-treated rats after H/R. Hepatic oxidative (4-hydroxynonenal) and nitrosative (3-nitrotyrosine) stress, inflammatory markers (serum IL-6 and hepatic infiltration with polymorphonuclear leukocytes), and actin cytoskeleton rearrangements were decreased after simvastatin pretreatment compared with vehicle-treated rats after H/R. Simvastatin increased eNOS and heme oxygenase 1 expression and eNOS activation. Expression of Rho/Rho-kinase and myosin phosphatase targeting subunit, Thr-MYPT1, a marker for Rho-kinase activity, decreased after simvastatin treatment compared with vehicle-treated rats after H/R. Simvastatin pretreatment exerts beneficial effects in this model of acute inflammation by supporting protective mechanisms that are important for hepatic microcirculation after H/R. PMID- 19997051 TI - Prior exercise alters responses to hemorrhage. AB - Traumatic injuries often occur to individuals while exercising. The author sought to determine whether exercise before injury resulting in hemorrhage would alter cardiovascular, metabolic, and neuroendocrine responses. Fifteen chronically instrumented splenectomized immature female swine were trained to run on a treadmill at 70% of maximum heart rate for 60 min. In six swine, responses to exercise were evaluated and found to return to baseline after 60 min of recovery. Swine were then randomly assigned to exercise (n = 7) or rest (n = 8) followed by hemorrhage of 25 mL/kg for 60 min then observed for an additional 60 min. The decrease in mean arterial pressure (MAP) was less after exercise, 26 +/- 9 mmHg compared with 49 +/- 2 mmHg with rest, with the difference sustained during the posthemorrhage period. Cardiac output decreased similarly in both groups. Posthemorrhage lactate and glucose concentrations were lower in exercise. The increase in plasma epinephrine was reduced in exercise, with significantly lower levels in epinephrine and norepinephrine noted posthemorrhage. Vasopressin levels and plasma renin activity were not different. In response to hemorrhage after exercise, blood pressure is better maintained although catecholamine levels were reduced, suggesting increased adrenoreceptor sensitivity. In addition, indices of increased glucose utilization and correction of lactate acidosis support a metabolic shift after exercise. Prior exercise alters responses to hemorrhage that mask the extent of hypovolemia and should be considered in the initial evaluation of a patient with hemorrhage caused by traumatic injuries. PMID- 19997052 TI - Tracking central hypovolemia with ecg in humans: cautions for the use of heart period variability in patient monitoring. AB - Heart period variability (HPV) metrics have been suggested for use in medical monitoring of trauma patients. This study sought to ascertain the use of various HPV metrics in tracking central blood volume during simulated hemorrhage in individual humans. One hundred one healthy nonsmoking volunteers (58 men, 43 women) were instrumented for continuous measurement of electrocardiogram and beat by-beat finger arterial blood pressure. Stroke volume (SV) was estimated from the arterial pulse wave and used to reflect central blood volume. Progressive lower body negative pressure (LBNP) was applied in 5-min stages until the onset of impending hemodynamic decompensation (systolic blood pressure <70 mmHg and/or presyncopal symptoms). HPV was assessed with analysis of R-to-R intervals using both linear (time and frequency domains) and nonlinear (e.g., complexity, fractality) methods. Application of increasing LBNP caused progressive reductions of SV, whereas arterial pressures changed only minimally and late. Group LBNP stage means for each HPV metric changed progressively and were strongly correlated with the mean decrease in SV (|r| > or = 0.87). To ascertain the utility of the HPV metrics to track individual responses to central hypovolemia, the difference scores for each HPV metric were correlated at each successive LBNP level, with percentage change in SV at the subject level. This cross-correlation of difference scores revealed that none of the HPV metrics showed strong and consistent correlations (|r| < or = 0.49) with percentage change in SV across successive LBNP levels. Although aggregate group mean values for HPV metrics are well correlated with SV changes during central hypovolemia, these metrics are less reliable when tracking individual reductions in central volume during LBNP. HPV metrics, therefore, may not be useful in monitoring hemorrhagic injuries in individual patients. PMID- 19997053 TI - Burn trauma induces early HMGB1 release in patients: its correlation with cytokines. AB - High-mobility group box protein 1 (HMGB1) is a nuclear protein that may be released actively from monocytes and macrophages or passively from necrotic or damaged cells. Several experimental data suggest that burn injury is accompanied by elevated plasma HMGB, but there are only few data available about its changes in burned patients. The aim of this study was to follow the time course and the prognostic value of plasma HMGB1 and cytokine changes in patients with severe burn injury affecting more than 10% of body surface area (n = 26). Blood samples were taken on admission and on the following 5 days. Plasma HMGB1 concentration was measured by the enzyme-linked immunosorbent assay method, whereas IL-6, IL-8, and IL-10 were assayed by the cytometric bead array kit. The HMGB1 and IL-10 concentrations were elevated on admission and gradually decreased thereafter. Significant differences were observed between survivors and nonsurvivors in HMGB1 (P < 0.01) and IL-10 (P < 0.001) concentrations on admission with higher levels in nonsurvivors. IL-6 and IL-8 started to increase markedly from day 2. Positive correlation (r = 0.669, P < 0.01) was found between burned body surface and HMGB1 on admission. Receiver operating characteristic analysis of data on admission showed that at a level of 16 ng/mL, HMGB1 indicated lethality, with 75.0% sensitivity and 85.7% specificity. Using the cutoff level of 14 pg/mL, IL-10 predicted intensive care unit mortality, with 85.7% sensitivity and 84.2% specificity. Very early HMGB1 and IL-10 release may have an important impact on the immune function of patients after burn trauma. PMID- 19997054 TI - Levosimendan increases portal blood flow and attenuates intestinal intramucosal acidosis in experimental septic shock. AB - It has been proposed that vasodilatory therapy may increase microcirculatory blood flow and improve tissue oxygenation in septic shock. The authors aimed to evaluate the effects of levosimendan in systemic and splanchnic hemodynamics in a porcine model of septic shock in a randomized animal controlled study. This study was performed in an animal research facility in a university hospital. Anesthetized pigs were monitored with a pulmonary artery catheter and an ultrasonic blood flow probe in the portal vein for measurement of systemic and portal blood flows and with a tonometer placed in the small intestine for measurement of the intramucosal-arterial PCO2 gap. Three groups of pigs were studied: nonseptic (n = 7), septic (n = 7), and septic treated with levosimendan (n = 7). Levosimendan was administered i.v. at t = -10 min (200 microg/kg in i.v. bolus followed by 200 microg/kg per h). Sepsis was induced at t = 0 min by the administration of live Escherichia coli. Vascular reactivity was tested by the hemodynamic response to noradrenaline. Levosimendan markedly attenuated the sepsis-induced increase in pulmonary vascular resistance, decrease in portal/systemic blood flow, oliguria, impairment in oxygenation, hyperkalemia, and the widened intramucosal-arterial PCO2 gap. Systemic blood pressure and vascular resistance did not differ as compared with the septic untreated group. Responses to noradrenaline significantly improved in animals treated with levosimendan. Treatment with levosimendan in this animal model of sepsis attenuated pulmonary vasoconstriction and improved portal blood flow, intestinal mucosal oxygenation, pulmonary function, and vascular reactivity. PMID- 19997055 TI - Mitochondrial DNA is released by shock and activates neutrophils via p38 map kinase. AB - Bacterial DNA (bDNA) can activate an innate-immune stimulatory "danger" response via toll-like receptor 9 (TLR9). Mitochondrial DNA (mtDNA) is unique among endogenous molecules in that mitochondria evolved from prokaryotic ancestors. Thus, mtDNA retains molecular motifs similar to bDNA. It is unknown, however, whether mtDNA is released by shock or is capable of eliciting immune responses like bDNA. We hypothesized shock-injured tissues might release mtDNA and that mtDNA might act as a danger-associated molecular pattern (or "alarmin") that can activate neutrophils (PMNs) and contribute to systemic inflammatory response syndrome. Standardized trauma/hemorrhagic shock caused circulation of mtDNA as well as nuclear DNA. Human PMNs were incubated in vitro with purified mtDNA or nuclear DNA, with or without pretreatment by chloroquine (an inhibitor of endosomal receptors like TLR9). Neutrophil activation was assessed as matrix metalloproteinase (MMP) 8 and MMP-9 release as well as p38 and p44/42 mitogen activated protein kinase (MAPK) phosphorylation. Mitochondrial DNA induced PMN MMP-8/MMP-9 release and p38 phosphorylation but did not activate p44/42. Responses were inhibited by chloroquine. Nuclear DNA did not induce PMN activation. Intravenous injection of disrupted mitochondria (mitochondrial debris) into rats induced p38 MAPK activation and IL-6 and TNF-alpha accumulation in the liver. In summary, mtDNA is released into the circulation by shock. Mitochondrial DNA activates PMN p38 MAPK, probably via TLR9, inducing an inflammatory phenotype. Mitochondrial DNA may act as a danger-associated molecular pattern or alarmin after shock, contributing to the initiation of systemic inflammatory response syndrome. PMID- 19997056 TI - Direct cardiac effects of dobutamine, dopamine, epinephrine, and levosimendan in isolated septic rat hearts. AB - In septic patients, myocardial depression-the so-called septic cardiomyopathy needing inotropic support is common. The aim of this study was to compare the dose-responsive electrophysiological and mechanical properties concerning metabolic effects of clinically available inotropic agents in an isolated septic rat heart model. After 20 h of incubation, both sham-operated and septic (cecal ligation and single puncture) hearts from male Wistar rats (n = 64) were isolated and received dobutamine, dopamine, epinephrine, or levosimendan at concentrations of 10 to 10 M. Electrophysiological, mechanical, and metabolic properties were measured, and the myocardial oxygen supply-demand ratio and cardiac efficiency were calculated. With the exception of levosimendan, all of the drugs tested showed dose-dependent, significantly positive changes in chronotropy, inotropy, and lusitropy in all hearts. Maximum increases in septic hearts were dose dependent and were ordered as follows: epinephrine > dopamine > dobutamine >>> levosimendan. These increases in cardiac performance were accompanied by a decrease in the myocardial oxygen supply-demand ratio. However, cardiac efficiency was significantly improved in the epinephrine-treated septic hearts. With the drug-induced increase in cardiac performance, the myocardial oxygen supply-demand ratio decreased proportionally in the epinephrine-, dobutamine-, and dopamine-treated septic hearts. However, epinephrine showed the most favorable results with regard to cardiac efficiency, and levosimendan showed no beneficial effect in septic hearts with regard to efficiency in this study. PMID- 19997057 TI - Activation of peroxisome proliferator-activated receptor-beta/delta attenuates myocardial ischemia/reperfusion injury in the rat. AB - Peroxisome proliferator-activated receptor-beta/delta (PPAR-beta/delta) is a transcription factor that belongs to the PPAR nuclear hormone receptor family. There is little information about the effects of the immediate administration of specific ligands of PPAR-beta/delta (e.g., GW0742) in animal models of myocardial I/R injury. Using a rat model of regional myocardial I/R in vivo, we have investigated the effects of immediate administration of GW0742 on myocardial infarct size. Male Wistar rats were subjected to 25 min of regional ischemia followed by 2 h of reperfusion and treated with GW0742 (3, 30, or 300microg/kg i.v. given at 30 min before ischemia and again at the start of reperfusion). Higher doses (30 or 300 microg/kg i.v.) of GW0742 caused a reduction in infarct size, whereas the lowest dose used was not effective. The degree of cardioprotection was similar when GW0742 (30 microg/kg i.v.) was given on reperfusion alone. The reduction in infarct size afforded by GW0742 was not reduced by the competitive irreversible PPAR-alpha antagonist GW6471 (1 mg/kg i.v., 15 min before ischemia). GW0742 (30 microg/kg i.v.) reduced the I/R-induced (a) decrease in the phosphorylation of Akt and glycogen synthase kinase-3beta, (b) nuclear translocation of the p65 subunit of nuclear factor-kappaB (activation of nuclear factor-kappaB), and (c) increase in the expression of iNOS and cyclooxygenase-2. Thus, immediate administration of the PPAR-beta/delta ligand GW0742 during reperfusion reduces myocardial infarct size in the rat by a mechanism that may involve inhibition of the activity of glycogen synthase kinase 3beta secondary to activation of the Akt pathway. PMID- 19997058 TI - The independent association between serum uric acid and graft outcomes after kidney transplantation. AB - BACKGROUND: Improving long-term outcomes of kidney transplantation depends on identifying novel risk factors that lead to poor outcomes. We sought to evaluate the predictive value of mean uric acid (UA) level during the first 6 months posttransplant for graft survival and function. METHODS: Two hundred twelve recipients of living donor kidneys transplanted during January 2000 to December 2001 were included. The study outcome included graft and patient survival and graft function at 1 year posttransplant. Regression models were used to adjust for the confounding variables including graft function during first 6 months. RESULTS: During 68.3 + or - 27.2 months follow-up, UA level (mg/dL) and hyperuricemia (n=45) were associated with graft loss (hazard ratio [HR]=1.26, P=0.026, 95% confidence interval [CI]=1.03-1.53, and HR=1.92, P=0.029, 95% CI=1.1 3.4, respectively) independent of graft function and other confounders. UA also seemed to be associated with risk of death with borderline significance (HR=1.2, P=0.096, 95% CI=0.97-1.46). Examining the predictive value for graft function, UA level and hyperuricemia were independent predictors of 1-year serum creatinine (beta=0.10, P=0.013, 95% CI=0.02-0.18, and beta=0.25, P<0.04, 95% CI=0.01-0.49, respectively). Similarly, both were associated with 1-year estimated glomerular filtration rate (beta=-3.9, P<0.001, 95% CI=-5.7 to -1.5 for UA, and beta=-7.6, P<0.02, 95% CI=-13.6 to -1.5 for hyperuricemia). Notably, these associations were all independent of renal function during first 6 months. CONCLUSION: The results of this study suggest that mean UA level during the first 6 months posttransplant is an independent predictor of long-term graft survival and short-term graft function. Further investigations are needed to evaluate its causal association with chronic allograft injury and cardiovascular disease. PMID- 19997059 TI - Quantitative short-tandem repeat analysis of recipient-derived cells as an additional tool for diagnosing cardiac allograft rejection. AB - BACKGROUND: Diagnosis of cardiac allograft rejection is currently based on histologic exploration of endomyocardial biopsies. Moderate interobserver reproducibility in the estimation of number and distribution of inflammatory cells leads to disagreements in the assignment of rejection grades. Short-tandem repeat (STR) analysis is routinely used after hematopoietic stem-cell transplantation to determine the proportions of donor cells. We compared the amount of recipient-derived cells with the histopathologic grade of rejection in cardiac allografts to determine whether this method might be useful for the assessment of rejection episodes. METHODS: One hundred forty-three endomyocardial biopsies from 18 patients were investigated for the percentage of recipient derived cell content by polymerase chain reaction-based STR analysis and correlated with rejection grades determined according to the International Society for Heart and Lung Transplantation grading system. Y-chromosome chromogene in situ hybridization was performed in gender-mismatched (female-to male) heart transplants to explore the influence of cardiomyocyte cell chimerism. RESULTS: The mean percentages of recipient-derived cells associated with various degrees of rejection were 13% in grade 0, 24% in grade 1A, 29% in grade 1B, 35% in grade 2, and 50% in grade > or =3A. Samples lacking signs of rejection (grade 0) had a significantly lower (P<0.001) amount of recipient-derived cells than higher degrees of rejections. Chromogene in situ hybridization analysis revealed that the recipient-derived cells were mainly inflammatory. CONCLUSIONS: The results of STR-analysis indicate that rejection is correlated with a higher proportion of recipient-derived cells. This assessment is observer independent and may thus represent an additional diagnostic tool for the assessment of rejection and management of immunosuppressive treatment. PMID- 19997060 TI - Predictors of survival following liver transplantation in infants: a single center analysis of more than 200 cases. AB - BACKGROUND: Infants (<12 months) who require liver transplantation (LTx) represent a particularly challenging and understudied group of patients. METHODS: This retrospective study aimed to describe a large single-center experience of infants who received isolated LTx, illustrate important differences in infants versus older children, and identify pretransplant factors which influence survival. More than 25 pre-LTx demographic, laboratory, and operative variables were analyzed using the Log-rank test and Cox proportional hazards model. RESULTS: Between 1984 and 2006, 216 LTx were performed in 186 infants with a mean follow-up time of 62 months. Median age at LTx was 9 months, the majority had cholestatic liver disease, were hospitalized pre-LTx, and received whole grafts. Leading indications for re-LTx (n=30) included vascular complications (43%) and graft nonfunction (40%), whereas leading causes of death were sepsis and multiorgan failure. One-, 5-, and 10-year graft and patient survivals were 75%/72%/68% and 79%/77%/75%, respectively. Relative to older pediatric recipients, infants had worse overall patient survival (P=0.05). The following were significant univariate predictors of graft loss: age less than 6 months and reduced cadaveric grafts; and of patient loss: age less than 6 months, calculated CrCl less than 90, pre-LTx hospitalization, pre-LTx mechanical ventilation, repeat LTx, infants transplanted for reasons other than cholestatic liver disease, and patients transplanted between 1984 and 1994. CONCLUSIONS: Long-term outcomes for infants undergoing LTx are excellent and have improved over time. As the largest, single-center analysis of LTx in infants, this study elucidates a unique set of predictors that can aid in medical decision making. PMID- 19997061 TI - Retro-orbital injection in adult zebrafish. AB - Drug treatment of whole animals is an essential tool in any model system for pharmacological and chemical genetic studies. Intravenous (IV) injection is often the most effective and noninvasive form of delivery of an agent of interest. In the zebrafish (Danio rerio), IV injection of drugs has long been a challenge because of the small vessel diameter. This has also proved a significant hurdle for the injection of cells during hematopoeitic stem cell transplantation. Historically, injections into the bloodstream were done directly through the heart. However, this intra-cardiac procedure has a very high mortality rate as the heart is often punctured during injection leaving the fish prone to infection, massive blood loss or fatal organ damage. Drawing on our experience with the mouse, we have developed a new injection procedure in the zebrafish in which the injection site is behind the eye and into the retro-orbital venous sinus. This retro-orbital (RO) injection technique has been successfully employed in both the injection of drugs in the adult fish as well as transplantation of whole kidney marrow cells. RO injection has a much lower mortality rate than traditional intra-cardiac injection. Fish that are injected retro-orbitally tend to bleed less following injection and are at a much lower risk of injury to a major organ like the heart. Further, when performed properly, injected cells and/or drugs quickly enter the bloodstream allowing compounds to exert their effect on the whole fish and kidney cells to easily home to their niche. Thus, this new injection technique minimizes mortality while allowing efficient delivery of material into the bloodstream of adult fish. Here we exemplify this technique by retro-orbital injection of Tg(globin:GFP) cells into adult casper fish as well as injection of a red fluorescent dye (dextran, Texas Red ) into adult casper fish. We then visualize successful injections by whole animal fluorescence microscopy. PMID- 19997062 TI - Generation of induced pluripotent stem cells by reprogramming human fibroblasts with the stemgent human TF lentivirus set. AB - In 2006, Yamanaka and colleagues first demonstrated that retrovirus-mediated delivery and expression of Oct4, Sox2, c-Myc and Klf4 is capable of inducing the pluripotent state in mouse fibroblasts.(1) The same group also reported the successful reprogramming of human somatic cells into induced pluripotent stem (iPS) cells using human versions of the same transcription factors delivered by retroviral vectors.(2) Additionally, James Thomson et al. reported that the lentivirus-mediated co-expression of another set of factors (Oct4, Sox2, Nanog and Lin28) was capable of reprogramming human somatic cells into iPS cells.(3) iPS cells are similar to ES cells in morphology, proliferation and the ability to differentiate into all tissue types of the body. Human iPS cells have a distinct advantage over ES cells as they exhibit key properties of ES cells without the ethical dilemma of embryo destruction. The generation of patient-specific iPS cells circumvents an important roadblock to personalized regenerative medicine therapies by eliminating the potential for immune rejection of non-autologous transplanted cells. Here we demonstrate the protocol for reprogramming human fibroblast cells using the Stemgent Human TF Lentivirus Set. We also show that cells reprogrammed with this set begin to show iPS morphology four days post transduction. Using the Stemolecule Y27632, we selected for iPS cells and observed correct morphology after three sequential rounds of colony picking and passaging. We also demonstrate that after reprogramming cells displayed the pluripotency marker AP, surface markers TRA-1-81, TRA-1-60, SSEA-4, and SSEA-3, and nuclear markers Oct4, Sox2 and Nanog. PMID- 19997063 TI - Detergent-resistant microdomains mediate activation of host cell signaling in response to attaching-effacing bacteria. AB - Enterohemorrhagic Escherichia coli (EHEC) O157:H7 causes outbreaks of bloody diarrhea and the hemolytic-uremic syndrome. EHEC intimately adheres to epithelial cells, effaces microvilli and induces attaching-effacing (AE) lesions. Detergent resistant microdomains (lipid rafts) serve as membrane platforms for the recruitment of signaling complexes to mediate host responses to infection. The aim of this study was to define the role of lipid rafts in activating signal transduction pathways in response to AE bacterial pathogens. Epithelial cell monolayers were infected with EHEC (MOI 100:1, 3 h, 37 degrees C) and lipid rafts isolated by buoyant density ultracentrifugation. Phosphoinositide 3-kinase (PI3K) localization to lipid rafts was confirmed using PI3K and anti-caveolin-1 antibodies. Mice with cholesterol storage disease Niemann-Pick, type C were used as in vivo models to confirm the role of lipid rafts in mediating signaling response to AE organisms. In contrast to uninfected cells, PI3K was recruited to lipid rafts in response to EHEC infection. Metabolically active bacteria and cells with intact cholesterol-rich microdomains were necessary for the recruitment of second messengers to lipid rafts. Recruitment of PI3K to lipid rafts was independent of the intimin (eaeA) gene, type III secretion system, and production of Shiga-like toxins. Colonization of NPC(-/-) colonic mucosa by Citrobacter rodentium and AE lesion formation were both delayed, compared with wild-type mice infected with the murine-specific AE bacterial pathogen. C. rodentium-infected NPC(-/-) mice had reduced colonic epithelial hyperplasia (64+/ 8.251 vs 112+/-2.958 microm; P<0.05) and decreased secretion of IFN-gamma (17.6+/ 17.6 vs 71+/-26.3 pg/ml, P<0.001). Lipid rafts mediate host cell signal transduction responses to AE bacterial infections both in vitro and in vivo. These findings advance the current understanding of microbial-eukaryotic cell interactions in response to enteric pathogens that hijack signaling responses mediated through lipid rafts. PMID- 19997064 TI - Complement system becomes activated by the classical pathway in intracranial aneurysm walls. AB - Inflammation and activation of the complement system in the intracranial aneurysm (IA) wall predispose to IA rupture. We have previously shown that increased C5b-9 accumulation correlates with IA rupture and wall degeneration. To elucidate the underlying mechanisms, we investigated initiators and the pathway of complement activation in unruptured and ruptured IAs. Unruptured and ruptured IA wall samples were studied in parallel sections by immunohistochemical and immunofluorescence stainings for the location and relations of classical and alternative pathway complement components (C1q, C3b/iC3b, C3d, C4b/iC4b; n=35 and properdin, n=10), putative complement activators IgG (n=90), IgM, CRP and OxLDL (n=10), and complement activation endproduct C5b-9. Classical pathway components were seen in all IAs, and they were located mostly in the extracellular matrix. The early pathway complement components colocalized with each other, but were present in larger areas than C5b-9. The areas positive for complement component accumulation were significantly broader in ruptured than in unruptured IAs. The potential complement activators IgG, IgM, CRP and OxLDL were found mostly in the extracellular matrix and in partial overlap with C5b-9. Lipids were seen in Oil Red-O staining in colocalization with C5b-9. Complement becomes activated by the classical pathway in the IA wall. The activation appears to be induced by multiple factors, which, in addition to the traditional activators (immunoglobulins, CRP, OxLDL), could involve vascular pressure-induced tissue damage. Despite wide early pathway activation, the terminal pathway is focused on a distinct lipid-rich layer. The profile of the complement components and the association of C5b-9 with lipids in the extracellular matrix indicate a long-term chronic inflammatory process rather than an acute targeted inflammatory reaction. The observed pattern of complement activation may be the consequence of local stress-induced insufficiency of complement regulation in IA walls. PMID- 19997065 TI - Epstein-Barr virus facilitates the malignant potential of immortalized epithelial cells: from latent genome to viral production and maintenance. AB - Epstein-Barr virus (EBV) is closely associated with several malignancies, including nasopharyngeal carcinoma. To investigate the EBV activity in tumor development, we tried to establish a malignant model of EBV-infected cells in nude mice. On the basis of the Maxi-EBV system, a human embryonic kidney epithelial cell line (293) with a low malignant potential was used for a stable EBV genome infection. The derived cell line, termed 293-EBV, exhibited obvious morphological transformation and significantly increased growth ability, with the cell cycle redistributed. The clonability and tumorigenicity were also substantially accelerated. In 293-EBV cells, the expression level of the transcription factor NF-kappaB and JNK2 were upregulated. The result suggested that latent membrane protein 1 (LMP1) was an important viral protein responsible for the enhanced malignant potential. Matured and budding virus particles were observed in tumor tissues, confirming the spontaneous reactivation of EBV from latent genome to lytic cycle at the site of tumor development. Primary culture of tumor tissues showed two patterns about the EBV maintenance or not in newly grown cells, and this was dependent on the thickness of the planted tissues. Moreover, the tumor cells lost EBV genome easily when subcultured at low density. Our findings revealed the cell-to-cell contact mechanism, which was required for the EBV maintenance in the tumor cells during the expansion of EBV-infected cells. This mechanism might give an explanation to the phenomenon that EBV genome in epithelial tumor cells becomes easily lost during subculture in vitro. Our results provided further evidence of a function for EBV in the etiology of tumor development. PMID- 19997066 TI - The protective role of pregnane X receptor in lipopolysaccharide/D-galactosamine induced acute liver injury. AB - The pregnane X receptor (PXR) is a nuclear receptor transcription factor regulating drug-metabolizing enzymes and transporters that facilitate xenobiotic and endobiotic detoxification. Recent studies show that PXR is important in abrogating intestinal tissue damage. This study examines the role of PXR in lipopolysaccharide (LPS)/D-galactosamine (GalN)-induced acute liver injury using wild-type and PXR-null mice. LPS/GalN-treated PXR-null mice had greater increases of alanine transaminase (ALT), hepatocyte apoptosis, necrosis, and hemorrhagic liver injury than wild-type mice. LPS/GalN-mediated phosphorylation of JNK1/2 and ERK1/2 was differentially regulated in wild-type and PXR-null mice. Importantly, LPS/GalN-induced hepatic Stat3 survival signaling was impaired and early activation of Jak2 was delayed in PXR-null mice. Expression levels of pro survival proteins Bcl-xL and heme oxygenase-1 (HO-1), which are downstream of Stat3, were substantially lower in PXR-null than wild-type mouse livers after LPS/GalN treatment. Autophagy is also involved in LPS/GalN-induced liver injury. Lack of PXR resulted in a significant reduction of LC3B-I, -II as well as Beclin 1 protein levels after LPS/GalN treatment. In addition, PXR is implicated in hepatocytes homeostasis. Taken together, PXR is a critical hepatoprotective factor. Increases of LPS/GalN-induced hepatocyte apoptosis and liver injury in PXR-null mice are due to deregulated mitogen-activated protein (MAP) kinase activation as well as delayed Jak2/Stat3 activation, which lead to a compromise in defense mechanisms that involve Bcl-xL-, HO-1, and autophagy-mediated pathways. PMID- 19997067 TI - Building the sequence map of the human pan-genome. AB - Here we integrate the de novo assembly of an Asian and an African genome with the NCBI reference human genome, as a step toward constructing the human pan-genome. We identified approximately 5 Mb of novel sequences not present in the reference genome in each of these assemblies. Most novel sequences are individual or population specific, as revealed by their comparison to all available human DNA sequence and by PCR validation using the human genome diversity cell line panel. We found novel sequences present in patterns consistent with known human migration paths. Cross-species conservation analysis of predicted genes indicated that the novel sequences contain potentially functional coding regions. We estimate that a complete human pan-genome would contain approximately 19-40 Mb of novel sequence not present in the extant reference genome. The extensive amount of novel sequence contributing to the genetic variation of the pan-genome indicates the importance of using complete genome sequencing and de novo assembly. PMID- 19997068 TI - Origin and evolution of the adaptive immune system: genetic events and selective pressures. AB - The adaptive immune system (AIS) in mammals, which is centred on lymphocytes bearing antigen receptors that are generated by somatic recombination, arose approximately 500 million years ago in jawed fish. This intricate defence system consists of many molecules, mechanisms and tissues that are not present in jawless vertebrates. Two macroevolutionary events are believed to have contributed to the genesis of the AIS: the emergence of the recombination activating gene (RAG) transposon, and two rounds of whole-genome duplication. It has recently been discovered that a non-RAG-based AIS with similarities to the jawed vertebrate AIS - including two lymphoid cell lineages - arose in jawless fish by convergent evolution. We offer insights into the latest advances in this field and speculate on the selective pressures that led to the emergence and maintenance of the AIS. PMID- 19997069 TI - Sequencing technologies - the next generation. AB - Demand has never been greater for revolutionary technologies that deliver fast, inexpensive and accurate genome information. This challenge has catalysed the development of next-generation sequencing (NGS) technologies. The inexpensive production of large volumes of sequence data is the primary advantage over conventional methods. Here, I present a technical review of template preparation, sequencing and imaging, genome alignment and assembly approaches, and recent advances in current and near-term commercially available NGS instruments. I also outline the broad range of applications for NGS technologies, in addition to providing guidelines for platform selection to address biological questions of interest. PMID- 19997070 TI - The role of testosterone in erectile dysfunction. AB - Erectile dysfunction (ED) is a clinical disorder that results from a continuous spectrum of clinical factors, including physical illness (comprising the organic component of ED), reaction to stress (the intrapsychic component of ED) and relationship difficulties (the relationship component of ED). Testosterone clearly has a relevant role in all three causes of ED; the usefulness of this hormone in the treatment of ED has not, however, been completely clarified. The main physiological action of testosterone in the male sexual response is to regulate the timing of the erectile process as a function of sexual desire, thereby coordinating penile erection with sex. The link between ED, hypogonadism and underlying disorders (such as metabolic syndrome and type 2 diabetes mellitus) is nowadays well documented. The recognition of underlying disorders might be useful in motivating men with ED to improve their health-related lifestyle choices. Hence, patients with ED might be considered 'lucky', because their disorder offers the opportunity to undergo medical examinations to detect underlying disease. Both ED and hypogonadism are treatable conditions. A range of testosterone preparations are available for supplementation; their combination with phosphodiesterase 5 inhibitors might improve outcomes in some cases. PMID- 19997071 TI - Nutraceuticals and prostate cancer prevention: a current review. AB - Nutraceuticals are 'natural' substances isolated or purified from food substances and used in a medicinal fashion. Several naturally derived food substances have been studied in prostate cancer in an attempt to identify natural preventative therapies for this disease. Vitamin E, selenium, vitamin D, green tea, soy, and lycopene have all been examined in human studies. Other potential nutraceuticals that lack human data, most notably pomegranate, might also have a preventative role in this disease. Unfortunately, most of the literature involving nutraceuticals in prostate cancer is epidemiological and retrospective. The paucity of randomized control trial evidence for the majority of these substances creates difficulty in making clinical recommendations particularly when most of the compounds have no evidence of toxicity and occur naturally. Despite these shortcomings, this area of prostate cancer prevention is still under intense investigation. We believe many of these 'natural' compounds have therapeutic potential and anticipate future studies will consist of well-designed clinical trials assessing combinations of compounds concurrently. PMID- 19997072 TI - The association between obesity and anxiety disorders in the population: a systematic review and meta-analysis. AB - Anxiety disorders are the most prevalent mental disorders in developed countries. Obesity is hypothesized to be a risk factor for anxiety disorders but evidence supporting an association between these two conditions is not clear. The objectives of this paper were to systematically review the literature for a link between obesity and anxiety disorders in the general population, and to present a pooled estimate of association. We performed a systematic search for epidemiological articles reporting on obesity (explanatory variable) and anxiety disorders (outcome variable) in seven bibliographical databases. Two independent reviewers abstracted the data and assessed study quality. We found 16 studies (2 prospective and 14 cross-sectional) that met the selection criteria. Measures of effect from prospective data were mixed but cross-sectional evidence suggested a positive association between obesity and anxiety. The pooled odds ratio from cross-sectional studies was 1.4 (confidence interval: 1.2-1.6). Subgroup analyses revealed a positive association in men and women. Overall, a moderate level of evidence exists for a positive association between obesity and anxiety disorders. Questions remain regarding the role of obesity severity and subtypes of anxiety disorders. The causal relationship from obesity to anxiety disorders could not be inferred from current data; future etiologic studies are recommended. PMID- 19997074 TI - Stereotactic body radiation therapy: a novel treatment modality. AB - Stereotactic body radiation therapy (SBRT) involves the delivery of a small number of ultra-high doses of radiation to a target volume using very advanced technology and has emerged as a novel treatment modality for cancer. The role of SBRT is most important at two cancer stages-in early primary cancer and in oligometastatic disease. This modality has been used in the treatment of early stage non-small-cell lung cancer, prostate cancer, renal-cell carcinoma, and liver cancer, and in the treatment of oligometastases in the lung, liver, and spine. A large body of evidence on the use of SBRT for the treatment of primary and metastatic tumors in various sites has accumulated over the past 10-15 years, and efficacy and safety have been demonstrated. Several prospective clinical trials of SBRT for various sites have been conducted, and several other trials are currently being planned. The results of these clinical trials will better define the role of SBRT in cancer management. This article will review the radiobiologic, technical, and clinical aspects of SBRT. PMID- 19997073 TI - MGMT promoter methylation in malignant gliomas: ready for personalized medicine? AB - The DNA repair enzyme O(6)-methylguanine-DNA methyltransferase (MGMT) antagonizes the genotoxic effects of alkylating agents. MGMT promoter methylation is the key mechanism of MGMT gene silencing and predicts a favorable outcome in patients with glioblastoma who are exposed to alkylating agent chemotherapy. This biomarker is on the verge of entering clinical decision-making and is currently used to stratify or even select glioblastoma patients for clinical trials. In other subtypes of glioma, such as anaplastic gliomas, the relevance of MGMT promoter methylation might extend beyond the prediction of chemosensitivity, and could reflect a distinct molecular profile. Here, we review the most commonly used assays for evaluation of MGMT status, outline the prerequisites for standardized tests, and evaluate reasons for difficulties in reproducibility. We critically discuss the prognostic and predictive value of MGMT silencing, reviewing trials in which patients with different types of glioma were treated with various chemotherapy schedules, either up-front or at recurrence. Standardization of MGMT testing requires comparison of different technologies across laboratories and prospectively validated cut-off values for prognostic or predictive effects. Moreover, future clinical trials will need to determine, for each subtype of glioma, the degree to which MGMT promoter methylation is predictive or prognostic, and whether testing should become routine clinical practice. PMID- 19997075 TI - Innovations in the systemic therapy of prostate cancer. AB - Systemic therapy has become an increasingly important component of treatment of advanced prostate cancer. In the past decade, important innovations have been achieved in the development of novel systemic hormonal therapies for the salvage treatment of metastatic castrate-resistant disease. These improvements have been accompanied by the broadening of potential indications for chemotherapy in castrate-resistant metastatic disease and the use of chemotherapy as an adjunct to the treatment of locally extensive tumors. These changes have begun to lead to improved outcomes, but at the expense of novel patterns of late toxic effects. We review the key steps in the recent evolution of systemic therapy of prostate cancer. PMID- 19997077 TI - Effects of cardiac rehabilitation referral strategies on referral and enrollment rates. AB - Despite recommendations in clinical practice guidelines, evidence suggests that utilization of cardiac rehabilitation (CR) following indicated cardiac events is low. Referral strategies, such as automatic referral, have been advocated to improve CR utilization. In this Review, we evaluate the effects of referral strategies on rates of CR referral and enrollment. Referral strategies are categorized as 'automatic' (the use of electronic health records or systematic discharge order sets), as 'liaison' (discussions with allied health-care providers), or as 'other' (for example, the use of motivational letter to patients). The highest rates of CR referral have been achieved in studies implementing automatic referral orders, whereas the highest rates of CR enrollment have resulted from a combination of automatic and liaison methods. Overall, innovative referral strategies significantly increase CR utilization. While further investigation is needed, institutions should evaluate their CR referral practice in light of these findings. PMID- 19997079 TI - Paclitaxel inhibits killing by murine cytotoxic T lymphocytes in vivo but not in vitro. AB - To kill antigen-specific target cells (TCs), cytotoxic T lymphocytes (CTLs) reorganise their microtubule cytoskeleton to deliver lytic granules to the TCs. We used two drugs that stabilise microtubules, paclitaxel and peloruside, to determine how the stabilising microtubule network affects CTL function in vitro and in vivo. In vitro, neither paclitaxel nor peloruside inhibited antigen specific killing, lytic granule delivery to the cell surface, nor interferon gamma release by murine CTLs. In contrast, in an in vivo murine model of antigen induced killing, a single dose of paclitaxel had a significant inhibitory effect on killing by CTLs. Furthermore, the inhibitory effect of paclitaxel was not caused by specific deletion of the effector CTL population in drug-treated mice. The findings reveal that microtubule-stabilising drug treatment can lead to immediate impairment of CTL function without affecting lytic granule release. The results also suggest that patients undergoing taxane anti-cancer therapy may be impaired in their ability to fight infection before the anti-mitotic effects of paclitaxel are apparent. PMID- 19997078 TI - PPAR-gamma agonist inhibits Ang II-induced activation of dendritic cells via the MAPK and NF-kappaB pathways. AB - The renin-angiotensin system exerts a profound regulatory effect on the functional features of dendritic cells (DCs), thus suggesting a new target of angiotensin II (Ang II) action in the immune system. This study analyzed whether peroxisome proliferator-activated receptor-gamma (PPAR-gamma) activation in DCs regulated Ang II-induced activation of DCs and exploited the possible molecular mechanisms, especially focused on the signaling pathways of mitogen-activated protein kinase (MAPK) and nuclear factor-kappa B (NF-kappaB). Ang II stimulation of human monocyte-derived DCs resulted in an intermediate state of DC maturation and function via modulating the balance of the negative or positive regulation of the signaling pathways of extracellular regulated kinase (ERK), p38 MAPK and NF kappaB, but not c-Jun N-terminal kinase (JNK). Moreover, pretreatment of DCs with the PPAR-gamma agonist pioglitazone reverted these effects of Ang II on DCs via suppression of the MAPK and NF-kappaB signaling pathways at least in part. Collectively, our data support the notion that PPAR-gamma activation in human DCs inhibits the activation of DCs induced by Ang II, with which involves the regulation of MAPK and NF-kappaB signaling pathways. These findings may support the important role of these mediators in the regulation of DC-mediated inflammatory and immunologic processes. PMID- 19997076 TI - Taxanes: optimizing adjuvant chemotherapy for early-stage breast cancer. AB - Taxanes are among the most widely used chemotherapy agents for advanced breast cancer. Results are now available from 21 trials that randomly allocated nearly 36,000 women with early-stage breast cancer to receive first-generation taxane based adjuvant chemotherapy versus non-taxane-based adjuvant regimens. Three recent meta-analyses suggest that taxanes are beneficial in the adjuvant setting, irrespective of the patient's age, lymph-node involvement, hormone-receptor expression, and HER2 status. Nevertheless, the optimal role for taxanes in the adjuvant management of early-stage breast cancer remains controversial. We review the results of the first-generation taxane trials and discuss possible explanations for the differences observed in these studies, including variation in the 'strength' of anthracycline therapy in the control arms; suboptimal timing, dosing, or schedule of the taxane regimen; a masking effect of trials that included patients with relatively chemotherapy-insensitive luminal A disease; and decreased representation of the putative taxane-sensitive disease subset. Inclusion criteria for future clinical trials must be revised to account for the molecular heterogeneity of breast cancer and further optimize the role of adjuvant taxane therapy in early-stage disease. PMID- 19997080 TI - Pharmacogenetics of risperidone therapy in autism: association analysis of eight candidate genes with drug efficacy and adverse drug reactions. AB - Little has been reported on the factors, genetic or other, that underlie the variability in individual response, particularly for autism. In this study we simultaneously explored the effects of multiple candidate genes on clinical improvement and occurrence of adverse drug reactions, in 45 autistic patients who received monotherapy with risperidone up to 1 year. Candidate genes involved in the pharmacokinetics (CYP2D6 and ABCB1) and pharmacodynamics (HTR2A, HTR2C, DRD2, DRD3, HTR6) of the drug, and the brain-derived neurotrophic factor (BDNF) gene, were analysed. Using the generalized estimating equation method these genes were tested for association with drug efficacy, assessed with the Autism Treatment Evaluation Checklist, and with safety and tolerability measures, such as prolactin levels, body mass index (BMI), waist circumference and neurological adverse effects, including extrapyramidal movements. Our results confirm that risperidone therapy was very effective in reducing some autism symptoms and caused few serious adverse effects. After adjusting for confounding factors, the HTR2A c.-1438G>A, DRD3 Ser9Gly, HTR2C c.995G>A and ABCB1 1236C>T polymorphisms were predictors for clinical improvement with risperidone therapy. The HTR2A c. 1438G>A, HTR2C c.68G>C (p.C33S), HTR6 c.7154-2542C>T and BDNF c.196G>A (p.V66M) polymorphisms influenced prolactin elevation. HTR2C c.68G>C and CYP2D6 polymorphisms were associated with risperidone-induced increase in BMI or waist circumference. We thus identified for the first time several genes implicated in risperidone efficacy and safety in autism patients. Although association results require replication, given the small sample size, the study makes a preliminary contribution to the personalized therapy of risperidone in autism. PMID- 19997081 TI - A multistep validation process of biomarkers for preclinical drug development. AB - Biomarkers that can be measured in preclinical models in a high-throughput, reproducible manner offer the potential to increase the speed and efficacy of drug development. Development of therapeutic agents for many conditions is hampered by the limited number of validated preclinical biomarkers available to gauge pharmacoefficacy and disease progression, but the validation process for preclinical biomarkers has received limited attention. This report defines a five step preclinical biomarker validation process and applies the process to a case study of diabetic retinopathy. By showing that a gene expression panel is highly reproducible, coincides with disease manifestation, accurately classifies individual animals and identifies animals treated with a known therapeutic agent, a biomarker panel can be considered validated. This particular biomarker panel consisting of 14 genes (C1inh, C1s, Carhsp1, Chi3l1, Gat3, Gbp2, Hspb1, Icam1, Jak3, Kcne2, Lama5, Lgals3, Nppa, Timp1) can be used in diabetic retinopathy pharmacotherapeutic research, and the biomarker development process outlined here is applicable to drug development efforts for other diseases. PMID- 19997082 TI - Antipsychotic drugs upregulate lipogenic gene expression by disrupting intracellular trafficking of lipoprotein-derived cholesterol. AB - Antipsychotic drugs (APDs) have been reported to induce lipogenic genes. This has been proposed to contribute to their efficacy in treating schizophrenia and other psychiatric disorders, as well as the metabolic side effects often associated with these drugs. The precise mechanism for the lipogenic effects of APDs is unknown, but is believed to involve increased activation of the lipogenic transcription factors, such as sterol regulatory element binding proteins (SREBPs). In a series of experiments in a model cell line, we found that a panel of typical and atypical APDs inhibited transport of lipoprotein-derived cholesterol to the endoplasmic reticulum (ER), which houses the cholesterol homeostatic machinery. APDs belong to the class of cationic amphiphiles and as has been shown for other amphiphiles, caused lipoprotein-derived cholesterol to accumulate intracellularly, preventing it from being esterified in the ER and suppressing SREBP activation. APDs did not activate the liver X receptor, another transcription factor involved in lipogenesis. However, these drugs markedly reduced cholesterol synthesis. This paradoxical result indicates that the upregulation of SREBP-target genes by APDs may not translate to increased cellular cholesterol levels. In conclusion, we have determined that APDs disrupt intracellular trafficking and synthesis of cholesterol, which may have important clinical ramifications. PMID- 19997083 TI - Alternative-splicing forms of the major phase II conjugating UGT1A gene negatively regulate glucuronidation in human carcinoma cell lines. AB - The UDP-glucuronosyltransferase UGT1A gene is a major biotransformation gene involved in the metabolism of a vast array of molecules. Recently, we uncovered a new series of alternative spliced isoforms referred to as isoforms 2 or UGT1As_i2 that use an alternative exon 5 (5b). The function of such mRNAs and the corresponding 45 kDa proteins still remains unclear. Although devoid of glucuronosyltransferase activity, UGT1As_i2 are widely co-expressed with the enzymatically active and classical UGT1A isoforms (UGT1As_i1). In this study, we observed abundant signal in human colon tissue samples, predominantly along intestinal crypts. In human cells, UGT1A_i2 proteins are expressed in similar subcellular compartments as UGT1As_i1. Cellular properties of i2-spliced forms were then studied using synthetic small-interfering RNA (siRNA) in two human colon cancer cell lines that show a significant amount of exon 5a- and exon 5b containing mRNAs and that display enzymatic activities for UGT1As substrates. We observed that siRNA-mediated knockdown of endogenous i2 upregulates cellular glucuronidation activities by 120-170% (P<0.01) for all substrates tested. Functional data support a dominant-negative function for endogenous exon 5b spliced forms of UGT1A, hence potentially affecting in vivo glucuronidation capacity. This new regulatory strategy may ensure an additional mean to modulate cellular response to endo/xeno stimulus. PMID- 19997085 TI - ApoE genotyping as a progression-rate biomarker in phase II disease-modification trials for Alzheimer's disease. PMID- 19997084 TI - Ethical considerations in the collection of genetic data from critically ill patients: what do published studies reveal about potential directions for empirical ethics research? AB - Critical illness trials involving genetic data collection are increasingly commonplace and pose challenges not encountered in less acute settings, related in part to the precipitous, severe and incapacitating nature of the diseases involved. We performed a systematic literature review to understand the nature of such studies conducted to date, and to consider, from an ethical perspective, potential barriers to future investigations. We identified 79 trials enrolling 24 499 subjects. Median (interquartile range) number of participants per study was 263 (116.75-430.75). Of these individuals, 16 269 (66.4%) were Caucasian, 1327 (5.4%) were African American, 1707 (7.0%) were Asian Pacific Islanders and 139 (0.6%) were Latino. For 5020 participants (20.5%), ethnicity was not reported. Forty-eight studies (60.8%) recruited subjects from single centers and all studies examined a relatively small number of genetic markers. Technological advances have rendered it feasible to conduct clinical studies using high-density genome-wide scanning. It will be necessary for future critical illness trials using these approaches to be of greater scope and complexity than those so far reported. Empirical research into issues related to greater ethnic inclusivity, accuracy of substituted judgment and specimen stewardship may be essential for enabling the conduct of such trials. PMID- 19997087 TI - Wwp2 mediates Oct4 ubiquitination and its own auto-ubiquitination in a dosage dependent manner. AB - Transcription factor Oct4 plays critical roles in maintaining pluripotency and controlling lineage commitment of embryonic stem cells (ESCs). Our previous study indicates that Wwp2, a mouse HECT-type E3 ubiquitin ligase, ubiquitinates Oct4 and promotes its degradation in a heterologous system. However, roles of Wwp2 in regulating endogenous Oct4 protein levels as well as molecular characteristics of the function of Wwp2 have not been determined. Here, we report that Wwp2 plays an important role in Oct4 ubiquitination and degradation during differentiation of embryonal carcinoma cells (ECCs), although it does not appear to affect Oct4 protein levels in the undifferentiated ECCs and ESCs. Importantly, inhibition of Wwp2 expression by specific RNA interference elevates the Oct4 protein level, leading to attenuation in retinoid acid-induced activation of differentiation related marker genes. Mechanistically, Wwp2 catalyzes Oct4 poly-ubiquitination via the lysine 63 linkage in a dosage-dependent manner. Interestingly, Wwp2 also regulates its own ligase activity in a similar manner. Moreover, auto ubiquitination of Wwp2 occurs through an intra-molecular mechanism. Taken together, these results demonstrate a crucial role of Wwp2 in controlling endogenous Oct4 protein levels during differentiation processes of ECCs and suggest an interesting dosage-dependent mechanism for regulating the catalytic activity of the E3 ubiquitin ligase, Wwp2. PMID- 19997086 TI - The nuclear signaling of NF-kappaB: current knowledge, new insights, and future perspectives. AB - The nuclear factor-kappa B (NF-kappaB) transcription factor plays a critical role in diverse cellular processes associated with proliferation, cell death, development, as well as innate and adaptive immune responses. NF-kappaB is normally sequestered in the cytoplasm by a family of inhibitory proteins known as inhibitors of NF-kappaB (IkappaBs). The signal pathways leading to the liberation and nuclear accumulation of NF-kappaB, which can be activated by a wide variety of stimuli, have been extensively studied in the past two decades. After gaining access to the nucleus, NF-kappaB must be actively regulated to execute its fundamental function as a transcription factor. Recent studies have highlighted the importance of nuclear signaling in the regulation of NF-kappaB transcriptional activity. A non-Rel subunit of NF-kappaB, ribosomal protein S3 (RPS3), and numerous other nuclear regulators of NF-kappaB, including Akirin, Nurr1, SIRT6, and others, have recently been identified, unveiling novel and exciting layers of regulatory specificity for NF-kappaB in the nucleus. Further insights into the nuclear events that govern NF-kappaB function will deepen our understanding of the elegant control of its transcriptional activity and better inform the potential rational design of therapeutics for NF-kappaB-associated diseases. PMID- 19997089 TI - Cocal-pseudotyped lentiviral vectors resist inactivation by human serum and efficiently transduce primate hematopoietic repopulating cells. AB - Lentiviral vectors are established as efficient and convenient vehicles for gene transfer. They are almost always pseudotyped with the envelope glycoprotein of vesicular stomatitis virus (VSV-G) due to the high titers that can be achieved, their stability, and broad tropism. We generated a novel cocal vesiculovirus envelope glycoprotein plasmid and compared the properties of lentiviral vectors pseudotyped with cocal, VSV-G, and a modified feline endogenous retrovirus envelope glycoprotein (RD114/TR). Cocal-pseudotyped lentiviral vectors can be produced at titers as high as with VSV-G, have a broad tropism, and are stable, allowing for efficient concentration by centrifugation. Additionally, cocal vectors are more resistant to inactivation by human serum than VSV-G-pseudotyped vectors, and efficiently transduce human CD34(+) nonobese diabetic/severe combined immunodeficient (NOD/SCID) mouse-repopulating cells (SRCs), and long term primate hematopoietic repopulating cells. These studies establish the potential of cocal-pseudotyped lentiviral vectors for a variety of scientific and therapeutic gene transfer applications, including in vivo gene delivery and hematopoietic stem cell (HSC) gene therapy. PMID- 19997090 TI - RNAi-mediated CCR5 silencing by LFA-1-targeted nanoparticles prevents HIV infection in BLT mice. AB - RNA interference (RNAi)-mediated knockdown of gene expression offers a novel treatment strategy for human immunodeficiency virus (HIV) infection. However, the major hurdle for clinical use is a practical strategy for small interfering RNA (siRNA) delivery to the multiple immune cell types important in viral pathogenesis. We have developed a novel immunoliposome method targeting the lymphocyte function-associated antigen-1 (LFA-1) integrin expressed on all leukocytes and evaluated it for systemic delivery of siRNA in a humanized mouse model. We show that in vivo administration of the LFA-1 integrin-targeted and stabilized nanoparticles (LFA-1 I-tsNPs) results in selective uptake of siRNA by T cells and macrophages, the prime targets of HIV. Further, in vivo administration of anti-CCR5 siRNA/LFA-1 I-tsNPs resulted in leukocyte-specific gene silencing that was sustained for 10 days. Finally, humanized mice challenged with HIV after anti-CCR5 siRNA treatment showed enhanced resistance to infection as assessed by the reduction in plasma viral load and disease-associated CD4 T cell loss. This study demonstrates the potential in vivo applicability of LFA-1 directed siRNA delivery as anti-HIV prophylaxis. PMID- 19997091 TI - Complete genetic correction of ips cells from Duchenne muscular dystrophy. AB - Human artificial chromosome (HAC) has several advantages as a gene therapy vector, including stable episomal maintenance that avoids insertional mutations and the ability to carry large gene inserts including the regulatory elements. Induced pluripotent stem (iPS) cells have great potential for gene therapy, as such cells can be generated from the individual's own tissues, and when reintroduced can contribute to the specialized function of any tissue. As a proof of concept, we show herein the complete correction of a genetic deficiency in iPS cells derived from Duchenne muscular dystrophy (DMD) model (mdx) mice and a human DMD patient using a HAC with a complete genomic dystrophin sequence (DYS-HAC). Deletion or mutation of dystrophin in iPS cells was corrected by transferring the DYS-HAC via microcell-mediated chromosome transfer (MMCT). DMD patient- and mdx specific iPS cells with the DYS-HAC gave rise to differentiation of three germ layers in the teratoma, and human dystrophin expression was detected in muscle like tissues. Furthermore, chimeric mice from mdx-iPS (DYS-HAC) cells were produced and DYS-HAC was detected in all tissues examined, with tissue-specific expression of dystrophin. Therefore, the combination of patient-specific iPS cells and HAC-containing defective genes represents a powerful tool for gene and cell therapies. PMID- 19997092 TI - Factors associated with non-adherence to oral medication for inflammatory bowel disease: a systematic review. AB - OBJECTIVES: Adherence is generally associated with improved treatment outcomes. Risk factors for non-adherence must be understood to improve adherence. A systematic review was undertaken to determine which variables were consistently associated with non-adherence to oral medication in inflammatory bowel disease (IBD). METHODS: The databases EMBASE, Medline, and PsycINFO were searched for titles relating to adherence, medication, and IBD (1980-2008). Primary, quantitative studies were included if they concerned adult patients with IBD, measured adherence to oral medication, and measured characteristics associated with adherence. The resulting 17 papers were independently reviewed by two researchers who also assessed their quality according to pre-defined criteria. The main outcome was the frequency with which demographic, clinical, treatment, and psychosocial variables were found to be statistically significantly associated with non-adherence. RESULTS: Non-adherence rates ranged from 7 to 72%, with most studies reporting that 30-45% of patients were non-adherent. No demographic, clinical, or treatment variables were consistently associated with non-adherence. Psychological distress and patients' beliefs about medications were both related to non-adherence in four out of five studies, and doctor patient discordance was associated with non-adherence in two out of three studies. CONCLUSIONS: This is the largest review of factors associated with non adherence in IBD. Demographic, clinical, and treatment variables were not consistently associated with non-adherence. Psychological distress, patients' beliefs about medications, and doctor-patient discordance were associated with non-adherence. These findings call into question some of the conclusions of earlier reviews that did not take into account nonsignificant findings. Practical suggestions for gastroenterologists and future research are discussed. PMID- 19997093 TI - High serum creatinine in acute pancreatitis: a marker for pancreatic necrosis? AB - OBJECTIVES: High serum creatinine is a well-known unfavorable prognostic parameter in acute pancreatitis. Elevated creatinine at 48 h after admission was recently described as a marker for pancreatic necrosis. As pancreatic necrosis is a serious complication of acute pancreatitis and its identification by a simple single laboratory test would be very helpful, the aim of this study was to test that statement. METHODS: In a prospective multicenter study of 462 patients with a first attack of acute pancreatitis, serum creatinine was determined on admission, and at 24 and 48 h thereafter, and compared with the findings of contrast-enhanced computed tomography (CT) performed within 96 h of admission. RESULTS: Pancreatic necrosis was present in 62 (13%) of the patients. Serum creatinine levels (abnormal > or = 2 mg/dl) on admission and after 24 and 48 h were evaluated vs. the presence or absence of pancreatic necrosis. Sensitivity rates varied between 14 and 23%, specificity between 95 and 97%, positive predictive values between 41 and 50%, and negative predictive values between 87 and 89%. Receiver operating characteristic curves revealed an area under the curve of between 0.604 and 0.669. CONCLUSIONS: An elevated serum creatinine concentration at any time during the first 48 h of admission is not a marker for pancreatic necrosis in a first attack of acute pancreatitis. If serum creatinine is normal, necrotizing pancreatitis is unlikely, and contrast-enhanced CT need not be performed unless complications occur and/or the patient's condition deteriorates. PMID- 19997094 TI - Increased proteasome-mediated degradation of occludin in irritable bowel syndrome. AB - OBJECTIVES: Proteasome-mediated protein degradation may contribute to the regulation of intestinal inflammation. At the same time, low-grade inflammation and increased intestinal permeability seem to be involved in the pathophysiology of irritable bowel syndrome (IBS). Thus, we aimed to evaluate proteasome composition and activities in colonic mucosa of IBS patients and its putative pathogenic role. METHODS: Proteasome activities and proteasome subunit expression were measured in colonic mucosa of IBS, Crohn's disease (CD), and control patients by fluorometric assays and western blot, respectively. Expression of inhibitor of kappa B factor (IkappaB alpha) and occludin, a tight junction protein, was also evaluated in colonic biopsies. The degradation of recombinant occludin incubated with protein extracts from colonic mucosa was evaluated in the presence or absence of proteasome inhibitor, MG132. RESULTS: Proteasome trypsin like activity was increased in IBS patients compared with CD and controls, whereas chymotrypsin-like activity was upregulated in CD patients only. Caspase like activity was reduced both in IBS and CD patients. IkappaB alpha expression was similar between IBS and controls. In contrast, occludin expression was lower in IBS than in controls, but occludin mRNA level was similar. Protein extracts from IBS patients but not from controls degraded recombinant occludin (20% over 160 min), which was blocked by MG132. Although mast cell number was increased in IBS patients, no correlation was found between this number and proteasome alterations. CONCLUSIONS: Our study shows that proteasome alterations are present in the colonic mucosa of IBS patients and may contribute to the pathophysiology of IBS by increasing occludin degradation. PMID- 19997095 TI - Characteristics of reflux episodes and symptom association in patients with erosive esophagitis and nonerosive reflux disease: study using combined impedance pH off therapy. AB - OBJECTIVES: We sought to compare reflux and symptom association patterns in patients with nonerosive reflux disease (NERD), erosive esophagitis (EE), and in healthy volunteers (HVs). METHODS: Patients with EE and NERD underwent combined impedance-pH monitoring. Normal values were defined on the basis of previously collected data from 48 HVs. We evaluated distal esophageal acid exposure time (AET), number and type of reflux episodes (acid, nonacid), acid and bolus clearance times, proximal extension of reflux episodes, and symptom association probability (SAP). RESULTS: Distal AET (percentage time, pH<4) was higher (P<0.01) in 58 EE patients (median 7.4%, 25-75th percentile 4.2-9.9%) compared with 168 NERD patients (4.2% (1.2-6.4%)) and 48 HVs (0.7% (0.2-1.4%)). Patients with EE and NERD had a higher (P<0.01) number of acid reflux episodes compared with HVs (51 (37-66) vs. 34 (22-51) vs. 17 (8-31); P<0.05), but a similar number of nonacid reflux episodes (22 (15-39) vs. 23 (15-38) vs. 18 (14-26); P=NS). The percentage of reflux episodes reaching the proximal esophagus was higher (P<0.01) in EE patients (57% (45-73%)) than in NERD patients (45% (36-60%)) and HVs (33% (19-46%)). A positive SAP for heartburn or regurgitation was found in 161 of 168 (96%) NERD and 54 of 58 (93%) EE patients (P=NS). CONCLUSIONS: Acid reflux episodes, volume, and acid clearance are important factors in the pathogenesis of reflux-induced lesions. Nonacid reflux contributes less to esophageal mucosa damage, but is involved in the development of reflux symptoms in both NERD and EE patients. PMID- 19997096 TI - Are colonoscopy and bowel ultrasound useful for assessing response to short-term therapy and predicting disease outcome of moderate-to-severe forms of ulcerative colitis?: a prospective study. AB - OBJECTIVES: Mucosal healing has been proposed as an important sign of the efficacy of medical treatment of inflammatory bowel disease; however, direct evidence in ulcerative colitis (UC) is scarce. We evaluated the usefulness of colonoscopy and bowel ultrasound (US) as indexes of response to short-term therapy and as predictors of subsequent outcome in UC. METHODS: A total of 83 patients with moderate-to-severe UC were recruited; endoscopic and US severity was graded 0-3 at entry according to validated scores. Of the recruited patients, 74, who were clinically responsive to steroids, were followed up with repeated colonoscopy and bowel US at 3, 9, and 15 months from recruitment. Concordance between clinical, endoscopic, and US scores at various visits was determined by kappa statistics. Multiple unconditional logistic regression models were used to assess the predictivity of clinical, endoscopic, and US scores measured at 3 and 9 months on the development of endoscopic UC relapse within 15 months. RESULTS: A variable concordance was found over time between endoscopic and clinical score (weighted kappa between 0.38 and 0.95), with high and consistent concordance between endoscopic and US scores (weighted kappa between 0.76 and 0.90). On logistic regression analysis, moderate-to-severe endoscopic and US scores at 3 months were associated with a high risk of endoscopic activity at 15 months (odds ratio (OR): 5.2; 95% confidence interval (CI): 1.6-17.6 and OR: 9.1; 95% CI: 2.5 33.5, respectively). CONCLUSIONS: Bowel US may be used as a surrogate of colonoscopy in assessing the short-term response of severe forms of UC to therapy. Both US score and endoscopic score after 3 months of steroid therapy predict outcome of disease at 15 months. PMID- 19997097 TI - Tumor-associated angiogenesis and lymphangiogenesis correlate with progression of intrahepatic cholangiocarcinoma. AB - OBJECTIVES: Little is known about the function of tumor-associated neovascularization in the progression of intrahepatic cholangiocarcinoma (IHC). This study was conducted to evaluate the influence of tumor-associated angiogenesis and lymphangiogenesis on progression of IHC. METHODS: We analyzed tissue specimens of IHC (N=114) by immunohistochemistry using the endothelial specific antibody CD31 and the lymphendothelial-specific antibody D2-40 and subsequently quantified microvessel density (MVD) and lymphatic microvessel density (LVD). To analyze the influence of tumor-associated angiogenesis and lymphangiogenesis on tumor progression, tumors were allocated according to mean MVD and LVD, respectively, into groups of "high" and "low" MVD and LVD, respectively, and various clinicopathological characteristics as well as recurrence and survival data were analyzed. RESULTS: IHC revealed an induction of tumor-associated angiogenesis and lymphangiogenesis. Tumors of "high" MVD displayed more frequently advanced primary tumor stages and multiple tumor nodes. Furthermore, patients with tumors of "high" MVD had an inferior curative resection rate and suffered more frequently from recurrence. A "high" LVD was correlated with increased nodal spread, and patients with "high" LVD tumors more frequently developed recurrence. In the univariate analysis, MVD and LVD revealed significant influence on survival, and MVD was identified as an independent prognostic factor for survival in the multivariate analysis. The 5-year survival of patients with "low" MVD tumors was 42.1%, compared with 2.2% in patients with "high" MVD tumors (P<0.001). CONCLUSIONS: This study suggests a critical function of tumor-associated angiogenesis and lymphangiogenesis for progression of IHC. Therefore, antiangiogenic and antilymphangiogenic approaches may have therapeutic potency in this tumor entity. PMID- 19997098 TI - Prejudice and truth about the effect of testosterone on human bargaining behaviour. AB - Both biosociological and psychological models, as well as animal research, suggest that testosterone has a key role in social interactions. Evidence from animal studies in rodents shows that testosterone causes aggressive behaviour towards conspecifics. Folk wisdom generalizes and adapts these findings to humans, suggesting that testosterone induces antisocial, egoistic, or even aggressive human behaviours. However, many researchers have questioned this folk hypothesis, arguing that testosterone is primarily involved in status-related behaviours in challenging social interactions, but causal evidence that discriminates between these views is sparse. Here we show that the sublingual administration of a single dose of testosterone in women causes a substantial increase in fair bargaining behaviour, thereby reducing bargaining conflicts and increasing the efficiency of social interactions. However, subjects who believed that they received testosterone-regardless of whether they actually received it or not-behaved much more unfairly than those who believed that they were treated with placebo. Thus, the folk hypothesis seems to generate a strong negative association between subjects' beliefs and the fairness of their offers, even though testosterone administration actually causes a substantial increase in the frequency of fair bargaining offers in our experiment. PMID- 19997099 TI - Pre-treatment with chemotherapy can enhance the antigenicity and immunogenicity of tumours by promoting adaptive immune responses. AB - BACKGROUND: Some cancer patients are immuno-compromised, and it has been long felt that immune-intervention is not compatible with standard chemotherapies. However, increasing evidence suggests that standard chemotherapy drugs may stimulate beneficial changes in both the immune system and tumour. METHODS: We have assessed the expression of human leucocyte antigen class 1 (HLA1) on tumour cells before and after chemotherapy agents (cyclophosphamide, oxaliplatin or gemcitabine). In addition, we show that chemotherapy-stressed tumour cells may release cytokines that enhance the interactions between dendritic cells (DCs) and T cells into growth media. RESULTS: Here we report that some chemotherapy agents can increase HLA1 expression in tumour cells, even when expression is low. Increases were associated with killing by cytotoxic T cells, which were negated by HLA1-blockade. Furthermore, T-cell function, as indicated by increased proliferation, was enhanced as supernatants derived from tumours treated with chemotherapy augmented DC-maturation and function. CONCLUSION: There is evidence that a facet of immune surveillance can be restored by appropriate chemotherapy agents. Also, tumours exposed to some chemotherapy may secrete cytokines that can mature DCs, which ultimately enhances T-cell responses. PMID- 19997100 TI - Mutation analysis of the MSMB gene in familial prostate cancer. AB - BACKGROUND: MSMB, a gene coding for beta-microseminoprotein, has been identified as a candidate susceptibility gene for prostate cancer (PrCa) in two genome-wide association studies (GWAS). SNP rs10993994 is 2 bp upstream of the transcription initiation site of MSMB and was identified as an associated PrCa risk variant. The MSMB protein is underexpressed in PrCa and it was previously proposed to be an independent marker for the recurrence of cancer after radical prostatectomy. METHODS: In this study, the coding region of this gene and 1500 bp upstream of the 5'UTR has been sequenced in germline DNA in 192 PrCa patients with family history. To evaluate the possible effects of these variants we used in silico analysis. RESULTS: No deleterious mutations were identified, however, nine new sequence variants were found, most of these in the promoter and 5'UTR region. In silico analysis suggests that four of these SNPs are likely to have some effect on gene expression either by affecting ubiquitous or prostate-specific transcription factor (TF)-binding sites or modifying splicing efficiency. INTERPRETATION: We conclude that MSMB is unlikely to be a familial PrCa gene and propose that the high-risk alleles of the SNPs in the 5'UTR effect PrCa risk by modifying MSMB gene expression in response to hormones in a tissue-specific manner. PMID- 19997101 TI - Reporting of prognostic studies of tumour markers: a review of published articles in relation to REMARK guidelines. AB - BACKGROUND: Poor reporting compromises the reliability and clinical value of prognostic tumour marker studies. We review articles to assess the reporting of patients and events using REMARK guidelines, at the time of guideline publication. METHODS: We sampled 50 prognostic tumour marker studies from higher impact cancer journals between 2006 and 2007. The inclusion criteria were cancer; focus on single biological tumour marker; survival analysis; multivariable analysis; and not gene array or proteomic data. Articles were assessed for the REMARK profile and other REMARK guideline items. We propose a reporting aid, the REMARK profile, motivated by the CONSORT flowchart. RESULTS: In 50 studies assessed for the REMARK profile, the number of eligible patients (56% of articles), excluded patients (54%) and patients in analyses (98%) was reported. Only 50% of articles reported the number of outcome events. In multivariable analyses, 54% and 30% of articles reported patient and event numbers for all variables. Of the studies, 66% used archival samples, indicating a potentially biased patient selection. Only 36% of studies reported clearly defined outcomes. CONCLUSIONS: Good reporting is critical for the interpretability and clinical applicability of prognostic studies. Current reporting of key information, such as the number of outcome events in all patients and subgroups, is poor. Use of the REMARK profile would greatly improve reporting and enhance prognostic research. PMID- 19997102 TI - Birth characteristics and the risk of childhood rhabdomyosarcoma based on histological subtype. AB - BACKGROUND: Little is known about risk factors for childhood rhabdomyosarcoma (RMS) and the histology-specific details are rare. METHODS: Case-control studies formed by linking cancer and birth registries of California, Minnesota, New York, Texas and Washington, which included 583 RMS cases (363 embryonal and 85 alveolar RMS) and 57 966 randomly selected control subjects, were analysed using logistic regression. The associations of RMS (overall, and based on embryonal or alveolar histology) with birth weight across five 500 g categories (from 2000 to 4500 g) were examined using normal birth weight (2500-3999 g) as a reference. Large (>90th percentile) and small (<10th percentile) size for gestational age were calculated based on birth weight distributions in controls and were similarly examined. RESULTS: High birth weight increased the risk of embryonal RMS and RMS overall. Each 500 g increase in birth weight increased the risk of embryonal RMS (odds ratio (OR)=1.27, 95% confidence interval (CI)=1.14-1.42) and RMS overall (OR=1.18, 95% CI=1.09-1.29). Large size for gestational age also significantly increased the risk of embryonal RMS (OR=1.42, 95% CI=1.03-1.96). CONCLUSIONS: These data suggest a positive association between accelerated in utero growth and embryonal RMS, but not alveolar RMS. These results warrant cautious interpretation owing to the small number of alveolar RMS cases. PMID- 19997104 TI - Influence of omega-6 PUFA arachidonic acid and bone marrow adipocytes on metastatic spread from prostate cancer. AB - BACKGROUND: Prostate cancer (CaP) preferentially metastasises to the bone, and we have previously shown that the poly-unsaturated fatty acid (PUFA) arachidonic acid (AA) is a potent stimulator of CaP invasion. Here we present that AA promotes CaP invasion by inducing bone marrow adipocyte formation. METHODS: Boyden invasion-chamber assays assessed the ability of dietary oils, their PUFA components, and specific PUFA-loaded adipocytes to induce PC-3 invasion. Lipid transfer and metabolism was followed using deuterated AA and Fourier Transform Infrared spectroscopy (FTIR). RESULTS: Poly-unsaturated fatty acid constituents, but not their corresponding dietary oils, induced PC-3 invasion. PUFAs induce bone marrow adipocyte (BM-Ad) differentiation with AA inducing higher levels of BM-Ad differentiation, as compared with other PUFAs (3998+/-514.4 vs 932+/-265.8; P=0.00002), which stimulated greater PC-3 invasion than free AA (22 408.5+/-607.4 vs 16 236+/-313.9; P=0.01111) or adipocytes generated in the presence of other PUFAs. In bone marrow co-culture PC-3 and BM-Ad interactions result in direct uptake and metabolism of AA by PC-3 cells, destruction of the adipocyte and subsequent formation of a bone metastasis. CONCLUSION: The data supports the hypothesis that AA not only promotes CaP invasion, it also prepares the 'soil', making it more supportive for implantation and propagation of the migrating metastatic cell. PMID- 19997103 TI - Prognostic effect of activated EGFR expression in human colon carcinomas: comparison with EGFR status. AB - BACKGROUND: Evidence suggests that epidermal growth factor receptor (EGFR) activation status may better predict the clinical behaviour of colon cancers than does EGFR expression. However, the prognostic effect of phospho-EGFR in primary colon cancer remains undefined. METHODS: Phospho-EGFR (Tyr-1173) and EGFR expression were analysed by immunohistochemistry (IHC) in tissue microarrays of TNM stage II and III colon cancers from completed adjuvant therapy trials (n=388). Staining intensity was scored and correlated with clinicopathological variables, DNA mismatch repair (MMR) status, rates of cell proliferation (Ki-67), apoptosis (caspase-3), and patient survival. RESULTS: Phospho-EGFR expression was detected in 157 of 388 (40%) tumours, whereas EGFR was found in 214 of 361 (59%). Although phospho-EGFR was unrelated to clinicopathological variables, strong EGFR intensity was associated with higher tumour stage (P=0.03). Tumours overexpressing EGFR (P=0.0002) or phospho-EGFR (P=0.015) showed increased Ki-67, but not caspase-3 expression. Phospho-EGFR was not prognostic. EGFR intensity was associated with worse disease-free survival (DFS) (hazard ratio (HR): 1.21 (1.03, 1.41); P=0.019) and overall survival (OS) (HR: 1.19 (1.02, 1.39); P=0.028). Tumours expressing both EGFR and phospho-EGFR had similar survival as EGFR alone. Stage and lymph node number were prognostic for DFS and OS, and histological grade for OS. EGFR was an independent predictor of DFS (P=0.042) after adjustment for stage, histological grade, age, and MMR status. CONCLUSION: Phospho-EGFR and EGFR expression were associated with tumour cell hyperproliferation. Phospho-EGFR was not prognostic, whereas increased EGFR intensity was independently associated with poor DFS. PMID- 19997105 TI - Human papillomavirus infection and squamous cell carcinoma of the conjunctiva. AB - BACKGROUND: Squamous cell carcinoma of the conjunctiva (SCCC) is associated with HIV-related immunosuppression, but human papillomavirus virus (HPV) is also suspected to have a role. We carried out a case-control study to assess the role of cutaneous and mucosal HPV types in SCCC, conjunctival dysplasia, and their combination (SCCC/dysplasia) in Uganda. METHODS: We compared HPV prevalence in frozen biopsies from 94 SCCC cases (79 of whom were found to be HIV-positive), 39 dysplasia cases (34 HIV-positive), and 285 hospital controls (128 HIV-positive) having other eye conditions that required surgery. Highly sensitive PCR assays that detect 75 HPV types were used. Odds ratios (ORs) and 95% confidence intervals (CIs) were computed, adjusting for, or stratifying by age, sex, and HIV status. RESULTS: Cutaneous HPV types were detected in 45% of SCCC cases, 41% of dysplasia cases and 11% of controls. Human papillomavirus virus 5 and 8 were the most common types in SCCC, and most often occurred in combination with other types. Associations were observed between SCCC/dysplasia and detection of both single (OR=2.3; 1.2-4.4) and multiple (OR=18.3; 6.2-54.4) cutaneous HPV types, and were chiefly based on findings in HIV-positive patients. Cutaneous HPV infections were rarely observed among HIV-negative patients and the association with SCCC/dysplasia was not significant (OR=2.4; 0.6-9.6) among them. Squamous cell carcinoma of the conjunctiva/dysplasia risk and mucosal HPV types were not associated in either HIV-positive or HIV-negative patients. CONCLUSIONS: We detected cutaneous HPV types in nearly half of SCCC/dysplasia cases and often multiple types (HPV5 and 8 being most common). The role of HIV (confounder or strong enhancer of cutaneous HPV carcinogenicity) is still uncertain. PMID- 19997106 TI - Gene set enrichment analysis provides insight into novel signalling pathways in breast cancer stem cells. AB - BACKGROUND: Tumour-initiating cells (TICs) or cancer stem cells can exist as a small population in malignant tissues. The signalling pathways activated in TICs that contribute to tumourigenesis are not fully understood. METHODS: Several breast cancer cell lines were sorted with CD24 and CD44, known markers for enrichment of breast cancer TICs. Tumourigenesis was analysed using sorted cells and total RNA was subjected to gene expression profiling and gene set enrichment analysis (GSEA). RESULTS: We showed that several breast cancer cell lines have a small population of CD24(-/low)/CD44(+) cells in which TICs may be enriched, and confirmed the properties of TICs in a xenograft model. GSEA revealed that CD24( /low)/CD44(+) cell populations are enriched for genes involved in transforming growth factor-beta, tumour necrosis factor, and interferon response pathways. Moreover, we found the presence of nuclear factor-kappaB (NF-kappaB) activity in CD24(-/low)/CD44(+) cells, which was previously unrecognised. In addition, NF kappaB inhibitor dehydroxymethylepoxyquinomicin (DHMEQ) prevented tumourigenesis of CD24(-/low)/CD44(+) cells in vivo. CONCLUSION: Our findings suggest that signalling pathways identified using GSEA help to identify molecular targets and biomarkers for TIC-like cells. PMID- 19997108 TI - Regarding: 'Epoetin beta treatment in patients with cancer chemotherapy induced anaemia: the impact of initial haemoglobin and target haemoglobin levels on survival, tumour progression and thromboembolic events'. PMID- 19997107 TI - Pemphigus vulgaris antigen mRNA quantification for the staging of sentinel lymph nodes in head and neck cancer. AB - BACKGROUND: Molecular diagnosis has been proposed to enhance the intra-operative diagnosis of sentinel lymph node (SLN) invasion in head and neck squamous cell carcinoma (HNSCC). Although cytokeratin (CK) mRNA quantification with real-time reverse transcriptase-PCR (QRT-PCR) has produced encouraging results, the more discriminating markers remain to be identified. METHODS: Pemphigus vulgaris antigen (PVA), squamous cell carcinoma antigen (SCCA), and CK17 mRNA were quantified using QRT-PCR, and the results were compared with an extensive histopathological examination of the entire SLNs on 78 SLNs harvested from 22 patients with HNSCC. RESULTS: SCCA and CK17 quantification showed significantly higher mRNA values for macrometastases (MAs) than for either negative or isolated tumour cell (ITC) SLNs (P<0.01). Pemphigus vulgaris antigen allowed the discrimination of all MAs and micrometastases from both negative and ITC SLNs (P<0.001). For the neck staging of patients, considering metastatic vs non metastatic status, receiver-operating characteristic curve analysis found areas under the curve of 93.8, 97.9, and 100% for CK17, SCCA, and PVA, respectively. With PVA, a cutoff value of 562 copies per 100 ng of cDNA permitted the correct distinction between patients with positive as opposed to negative neck nodes in all cases. CONCLUSION: PVA seems to be a highly promising marker for accurate intra-operative SLN staging in HNSCC by QRT-PCR. PMID- 19997109 TI - Epoetin-beta treatment in patients with cancer chemotherapy-induced anaemia: the impact of initial haemoglobin and target haemoglobin levels on survival, tumour progression and thromboembolic events. AB - BACKGROUND: Epoetin-beta is used to treat patients with cancer undergoing chemotherapy to alleviate the symptoms of anaemia, reduce the risk of blood transfusions and improve quality of life (QoL). METHODS: This meta-analysis of all 12 randomised, controlled studies of epoetin-beta evaluated the impact of therapy at different Hb-initiation levels and to different target Hb levels on overall survival, tumour progression and thromboembolic events (TEE). An analysis of risk factors pre-disposing patients to TEEs under epoetin-beta therapy was also performed. A total of 2297 patients are included in the analysis. RESULTS: Analyses based on various Hb-initiation levels indicate no detrimental impact on survival (HR 0.99; 95% CI 0.70, 1.40) and a favourable impact on disease progression (HR 0.73; 95% CI 0.57, 0.94) when epoetin-beta was used within its licensed indication (Hb initiation < or = 10 g dl(-1)) or the EORTC recommended level of 11 g dl(-1). An increased risk of TEEs is seen for all Hb-initiation level strata and a detrimental impact on survival is seen when initiating epoetin beta therapy at Hb levels >11 g dl(-1). We observe no association between high target Hb levels (> or = 13 g dl(-1)) and an increased risk of mortality, disease progression or TEEs with epoetin-beta compared with control. CONCLUSION: The results of this analysis indicate that epoetin-beta therapy has no detrimental impact on survival or tumour progression when initiated at Hb levels up to 11 g dl(-1). Furthermore, there is no evidence to suggest that high Hb values achieved during epoetin-beta therapy are associated with an increased mortality, disease progression or TEE rate. PMID- 19997110 TI - Regarding: 'high-intensity-focused ultrasound in the treatment of primary prostate cancer: the first UK series'. PMID- 19997112 TI - Regarding: 'High-intensity-focused ultrasound in the treatment of primary prostate cancer: the first UK series'. PMID- 19997114 TI - Profiling of high-grade central osteosarcoma and its putative progenitor cells identifies tumourigenic pathways. PMID- 19997115 TI - The cerebellar microcircuit as an adaptive filter: experimental and computational evidence. AB - Initial investigations of the cerebellar microcircuit inspired the Marr-Albus theoretical framework of cerebellar function. We review recent developments in the experimental understanding of cerebellar microcircuit characteristics and in the computational analysis of Marr-Albus models. We conclude that many Marr-Albus models are in effect adaptive filters, and that evidence for symmetrical long term potentiation and long-term depression, interneuron plasticity, silent parallel fibre synapses and recurrent mossy fibre connectivity is strikingly congruent with predictions from adaptive-filter models of cerebellar function. This congruence suggests that insights from adaptive-filter theory might help to address outstanding issues of cerebellar function, including both microcircuit processing and extra-cerebellar connectivity. PMID- 19997116 TI - Inhibition of reactive astrocytes with fluorocitrate retards neurovascular remodeling and recovery after focal cerebral ischemia in mice. AB - Glial scarring is traditionally thought to be detrimental after stroke. But emerging studies now suggest that reactive astrocytes may also contribute to neurovascular remodeling. Here, we assessed the effects and mechanisms of metabolic inhibition of reactive astrocytes in a mouse model of stroke recovery. Five days after stroke onset, astrocytes were metabolically inhibited with fluorocitrate (FC, 1 nmol). Markers of reactive astrocytes (glial fibrillary acidic protein (GFAP), HMGB1), markers of neurovascular remodeling (CD31, synaptophysin, PSD95), and behavioral outcomes (neuroscore, rotarod latency) were quantified from 1 to 14 days. As expected, focal cerebral ischemia induced significant neurological deficits in mice. But over the course of 14 days after stroke onset, a steady improvement in neuroscore and rotarod latencies were observed as the mice spontaneously recovered. Reactive astrocytes coexpressing GFAP and HMGB1 increased in peri-infarct cortex from 1 to 14 days after cerebral ischemia in parallel with an increase in the neurovascular remodeling markers CD31, synaptophysin, and PSD95. Compared with stroke-only controls, FC-treated mice demonstrated a significant decrease in HMGB1-positive reactive astrocytes and neurovascular remodeling, as well as a corresponding worsening of behavioral recovery. Our results suggest that reactive astrocytes in peri-infarct cortex may promote neurovascular remodeling, and these glial responses may aid functional recovery after stroke. PMID- 19997117 TI - Overexpression of heat shock protein 27 reduces cortical damage after cerebral ischemia. AB - Heat shock protein 27 (HSP27) has a major role in mediating survival responses to a range of central nervous system insults, functioning as a protein chaperone, an antioxidant, and through inhibition of cell death pathways. We have used transgenic mice overexpressing HSP27 (HSP27tg) to examine the role of HSP27 in cerebral ischemia, using model of permanent middle cerebral artery occlusion (MCAO). Infarct size was evaluated using multislice T(2)-weighted anatomical magnetic resonance imaging (MRI) after 24 h. A significant reduction of 30% in infarct size was detected in HSP27tg animals compared with wild-type (WT) littermates. To gain some insight into the mechanisms contributing to cell death and its attenuation by HSP27, we monitored the effect of induction of c-jun and ATF3 on tissue survival in MCAO and their effects on the expression of endogenous mouse HSP25 and HSP70. It is important that, the c-jun induction seen at 4 h tended to be localized to regions that were salvageable in HSP27tg mice but became infarcted in WT animals. Our results provide support for the powerful neuroprotective effects of HSP27 in cerebral ischemia. PMID- 19997118 TI - Matrix metalloproteinase-9 mediates hypoxia-induced vascular leakage in the brain via tight junction rearrangement. AB - Blood-brain barrier (BBB) disruption, resulting from loss of tight junctions (TJ) and activation of matrix metalloproteinases (MMPs), is associated with edema formation in ischemic stroke. Cerebral edema develops in a phasic manner and consists of both vasogenic and cytotoxic components. Although it is contingent on several independent mechanisms, involving hypoxic and inflammatory responses, the single effect of prolonged hypoxia on BBB integrity in vivo was not addressed so far. Exposing mice to normobaric hypoxia (8% oxygen for 48 h) led to a significant increase in vascular permeability associated with diminished expression of the TJ protein occludin. Immunofluorescence studies revealed that hypoxia resulted in disrupted continuity of occludin and zonula occludens-1 (Zo 1) staining with significant gap formation. Hypoxia increased gelatinolytic activity specifically in vascular structures and gel zymography identified MMP-9 as enzymatic source. Treatment with an MMP inhibitor reduced vascular leakage and attenuated disorganization of TJ. Inhibition of vascular endothelial growth factor (VEGF) attenuated vascular leakage and MMP-9 activation induced by hypoxia. In conclusion, our data suggest that hypoxia-induced edema formation is mediated by MMP-9-dependent TJ rearrangement by a mechanism involving VEGF. Therefore, inhibition of MMP-9 might provide the basis for therapeutic strategies to treat brain edema. PMID- 19997119 TI - Simultaneous estimation of input functions: an empirical study. AB - In neuroreceptor mapping, methods for the estimation of distribution volume require determination of a metabolite-corrected arterial input function. In application, this may be accomplished by collecting arterial blood samples during scanning, adjusting these measurements according to a separate metabolite analysis, and then modeling the resulting concentration data. Although many groups do this routinely, it is invasive and requires considerable effort. Furthermore, both the plasma and the metabolite data are noisy, and thus estimation of kinetic parameters can be affected by this variability. One promising alternative to full-input function modeling is the simultaneous estimation (SIME) approach, in which kinetic parameters and common input function parameters are estimated using results obtained from several regions at once. We investigate the performance of this approach on data from four different radioligands, using various kinetic models, comparing the results with those obtained by estimation using full-input function modeling. Results indicate that SIME provides a promising alternative for all the radioligands considered. PMID- 19997120 TI - Using linked markers to estimate the genetic age of a volunteer population: a theoretical and empirical approach. AB - Volunteers deriving from unharvested seeds of a crop can lead to persistent feral populations and participate in genetic exchanges across the agro-ecosystem, both between crop varieties and between crops and their wild relatives. A first step to understand the importance of volunteers is to characterize their capacity to reproduce autonomously for several generations. For that purpose, we constructed and evaluated a maximum-likelihood method to estimate the genetic age of a population deriving from one of the most common field crop type: an F1-hybrid variety. The method estimates the number of reproduction cycles that occurred since the cultivation of that variety. It makes use of genotypic data at a number of linked microsatellite loci pairs, thus exploiting the recombination of parental haplotypes, which is expected to occur as the population is reproducing. Estimates with moderate bias and variance were found for a broad range of parameter values in simulations, and the method revealed robust to some deviations from the assumptions of the underlying model. We propose a specific procedure to test the hypothesis of persistence, that is has a given volunteer population experienced more than one cycle of reproduction since the F1-hybrid state? The method was applied to both an experimental and a natural sunflower volunteer population and revealed promising, considering these ideal case studies. Possible further developments toward more complex natural systems are discussed. PMID- 19997121 TI - Multiple introductions of divergent genetic lineages in an invasive fungal pathogen, Cryphonectria parasitica, in France. AB - The occurrence of multiple introductions may be a crucial factor in the successful establishment of invasive species, but few studies focus on the introduction of fungal pathogens, despite their significant effect on invaded habitats. Although Cryphonectria parasitica, the chestnut blight fungus introduced in North America and Europe from Asia during the 20th century, caused dramatic changes in its new range, the history of its introduction is not well retraced in Europe. Using 10 microsatellite loci, we investigated the genetic diversity of 583 isolates in France, where several introductions have been hypothesized. Our analyses showed that the seven most frequent multilocus genotypes belonged to three genetic lineages, which had a different and geographically limited distribution. These results suggest that different introduction events occurred in France. Genetic recombination was low among these lineages, despite the presence of the two mating types in each chestnut stand analysed. The spatial distribution of lineages suggests that the history of introductions in France associated with the slow expansion of the disease has contributed to the low observed rate of recombination among the divergent lineages. However, we discuss the possibility that environmental conditions or viral interactions could locally reduce recombination among genotypes. PMID- 19997122 TI - Contrasting relations between diversity of candidate genes and variation of bud burst in natural and segregating populations of European oaks. AB - Nucleotide diversity was assessed within nine candidate genes (in total 4.6 kb) for the time of bud burst in nine sessile oak (Quercus petraea) populations distributed in central and northern Europe. The sampled populations were selected on the basis of their contrasting time of bud burst observed in common garden experiments (provenance tests). The candidate genes were selected according to their expression profiles during the transition from quiescent to developing buds and/or their functional role in model plants. The overall nucleotide diversity was large (pi(tot)=6.15 * 10(-3); pi(silent)=11.2 * 10(-3)), but population differentiation was not larger than for microsatellites. No outlier single nucleotide polymorphism (SNP), departing from neutral expectation, was found among the total of 125 SNPs. These results contrasted markedly with the significant associations that were observed between the candidate genes and bud burst in segregating populations. Quantitative trait loci (QTLs) for bud burst were identified for 13 year*site seasonal observations in a cloned mapping pedigree. Nineteen QTLs were detected, and QTLs located on linkage groups 2, 5 and 9 contributed repeatedly to more than 12% of the phenotypic variation of the trait. Eight genes were polymorphic in the two parents of the pedigree and could be mapped on the existing genetic map. Five of them located within the confidence intervals of QTLs for bud burst. Interestingly, four of them located within the three QTLs exhibiting the largest contributions to bud burst. PMID- 19997123 TI - European and Polynesian admixture in the Norfolk Island population. AB - The Norfolk Island population in the South Pacific is primarily the product of recent admixture between a small number of British male and Polynesian female founders. We identified and genotyped 128 Ancestry Informative Markers (AIMs) spread across the autosomes, X/Y chromosomes and mitochondrial DNA genome, to explore and quantify the current levels of genetic admixture in the Norfolk Islanders. On the basis of autosomal AIMs, the population shows mean European and Polynesian ancestry proportions of 88 and 12%, respectively. However, there is a substantial variation between individuals ranging from total European ancestry to near total Polynesian origin. There is a strong correlation between individual genetic estimates of Polynesian ancestry and those derived from the extensive pedigree and genealogical records of Islanders. Also in line with historical accounts, there is a substantial asymmetry in the maternal and paternal origins of the Islanders with almost all Y-chromosomes of European origin whereas at least 25% of mtDNAs appear to have a Polynesian origin. Accurate knowledge of ancestry will be important in future attempts to use the Island population in admixture mapping approaches to find the genes that underlie differences in the risk to some diseases between Europeans and Polynesians. PMID- 19997124 TI - Genetics of species differences in sailfin and shortfin mollies. AB - Premating reproductive isolation is a strong barrier to hybridization in natural populations, but little is known about the genetic mechanisms that allow changes in mating signals to develop and whether different components of a mating signal can evolve in concert when sexual selection favors phenotypic associations between them. In this study, we report results suggesting that changes in a behavioural trait (courtship display) and multiple phenotypically associated morphological traits (dorsal fin characters and length of the gonopodium) have contributed to divergence in mating signals used by sailfin mollies. Through the use of reciprocal F1 and backcross hybrids, we show that morphological traits important in separating sailfin from shortfin molly species have a genetic basis and are inherited in an autosomal, additive manner. We also report significant associations between the size of certain morphological traits (length of the dorsal fin and length of the gonopodium) and the tendency of males to perform courtship displays or gonopodial thrusts. In particular, higher courtship display rates were associated with increased dorsal fin length but decreased gonopodium length, characteristics most similar to sailfin species. Such phenotypic associations between different components of a mating signal suggest that selective forces can act in concert on multiple aspects of the signal, hence, promoting divergence and speciation in sailfin mollies. PMID- 19997125 TI - Mechanisms and evolution of genomic imprinting in plants. AB - Genomic imprinting, the allele-specific expression of a gene dependent on its parent-of-origin, has independently evolved in flowering plants and mammals. In mammals and flowering plants, imprinting occurs in the embryo as well as in embryo-nourishing tissues, the placenta and the endosperm, respectively, and it has been suggested that imprinted genes control the nutrient flow from the mother to the offspring ('kinship theory'). Alternatively, imprinting might have evolved as a by-product of a defense mechanism destined to control transposon activity in gametes ('defense hypothesis'). Recent studies provide substantial evidence for the 'defense hypothesis' by showing that imprinted genes in plants are located in the vicinity of transposon or repeat sequences, suggesting that the insertion of transposon or repeat sequences was a prerequisite for imprinting evolution. Transposons or repeat sequences are silenced by DNA methylation, causing silencing of neighboring genes in vegetative tissues. However, because of genome wide DNA demethylation in the central cell, genes located in the vicinity of transposon or repeat sequences will be active in the central cell and the maternal alleles will remain unmethylated and active in the descendent endosperm, assuming an imprinted expression. Consequently, many imprinted genes are likely to have an endosperm-restricted function, or, alternatively, they have no functional role in the endosperm and are on the trajectory to convert to pseudogenes. Thus, the 'defense hypothesis' as well as 'kinship theory' together can explain the origin of genomic imprinting; whereas the first hypothesis explains how imprinting originates, the latter explains how imprinting is manifested and maintained. PMID- 19997126 TI - The genetic structure of populations of an invading pest fruit fly, Bactrocera tryoni, at the species climatic range limit. AB - Previous population genetic studies of the Queensland fruit fly, Bactrocera tryoni Froggatt (Diptera: Tephritidae), in its central range have shown barely detectable genetic differentiation across distances of almost 3000 km (F(st)=0.003). In this study, we investigated the genetic structuring of southern border populations of B. tryoni, in the region extending from the central population to the recently colonized southern range limit. The expectation was that marginal populations would be small, fragmented population sinks, with local adaptation limited by gene flow or drift. Unexpectedly, we found that the population at the southern extreme of the range was a source population, rather than a sink, for the surrounding region. This was shown by assignment testing of recent outbreaks in an adjoining quarantine area and by indirect migration estimates. Furthermore, populations in the region had formed a latitudinal cline in microsatellite allele frequencies, spanning the region between the central population and the southern range limit. The cline has formed within 250 generations of the initial invasion and appears stable between years. We show that there is restricted gene flow in the region and that effective population sizes are of the order of 10(2)-10(3). Although the cline may result from natural selection, neutral evolutionary processes may also explain our findings. PMID- 19997127 TI - Heritable artificial sex chromosomes in the medaka, Oryzias latipes. AB - Chromosomal sex determination is widely used by vertebrates, however, only two genes have been identified as master sex-determining genes: SRY/Sry in mammals and DMY in the teleost medaka. Transfer of both genes into genetically female (XX) individuals can induce male development. However, transgenic strains have not been established in both cases because of infertility of the transgenic founders in mammals and low germline transmission rates in medaka. In this study, we used a BAC clone containing DMY in a 117 kb genomic region and two types of fluorescent marker to establish two DMY-transgenic medaka strains. In these strains, exogenous DMY is completely linked to a male phenotype and early gonadal development is not different from that of the wild-type strain. Sex-linkage analysis showed that the exogenous DMY was located on linkage group (LG) 23 in one strain and on LG 5 in the other strain, whereas the sex chromosome in medaka is on LG 1. Real-time PCR analysis indicated that these strains have multiple copies of DMY and higher DMY expression levels than the wild-type strain. These results showed that LGs 23 and 5 function as sex chromosomes in the two strains, respectively. This is not only the first example of the artificial generation of heritable sex chromosomes in vertebrates but also the first evidence showing plasticity of homomorphic sex chromosomes. This plasticity appears to be a characteristic of lower vertebrates and the underlying cause of frequent sex chromosome switching, recently reported in several fish and frog species. PMID- 19997128 TI - Enhancer elements upstream of the SHOX gene are active in the developing limb. AB - Leri-Weill Dyschondrosteosis (LWD) is a dominant skeletal disorder characterized by short stature and distinct bone anomalies. SHOX gene mutations and deletions of regulatory elements downstream of SHOX resulting in haploinsufficiency have been found in patients with LWD. SHOX encodes a homeodomain transcription factor and is known to be expressed in the developing limb. We have now analyzed the regulatory significance of the region upstream of the SHOX gene. By comparative genomic analyses, we identified several conserved non-coding elements, which subsequently were tested in an in ovo enhancer assay in both chicken limb bud and cornea, where SHOX is also expressed. In this assay, we found three enhancers to be active in the developing chicken limb, but none were functional in the developing cornea. A screening of 60 LWD patients with an intact SHOX coding and downstream region did not yield any deletion of the upstream enhancer region. Thus, we speculate that SHOX upstream deletions occur at a lower frequency because of the structural organization of this genomic region and/or that SHOX upstream deletions may cause a phenotype that differs from the one observed in LWD. PMID- 19997131 TI - ["...and discussions increase the fear"]. PMID- 19997129 TI - Sensors and regulators of intracellular pH. AB - Protons dictate the charge and structure of macromolecules and are used as energy currency by eukaryotic cells. The unique function of individual organelles therefore depends on the establishment and stringent maintenance of a distinct pH. This, in turn, requires a means to sense the prevailing pH and to respond to deviations from the norm with effective mechanisms to transport, produce or consume proton equivalents. A dynamic, finely tuned balance between proton extruding and proton-importing processes underlies pH homeostasis not only in the cytosol, but in other cellular compartments as well. PMID- 19997132 TI - [Nobel Prize for mitotic clock]. PMID- 19997133 TI - [Aid for better health--role of Norway]. PMID- 19997130 TI - Unleashing formins to remodel the actin and microtubule cytoskeletons. AB - Formins are highly conserved proteins that have essential roles in remodelling the actin and microtubule cytoskeletons to influence eukaryotic cell shape and behaviour. Recent work has identified numerous cellular factors that locally recruit, activate or inactivate formins to bridle and unleash their potent effects on actin nucleation and elongation. The effects of formins on microtubules have also begun to be described, which places formins in a prime position to coordinate actin and microtubule dynamics. The emerging complexity in the mechanisms governing formins mirrors the wide range of essential functions that they perform in cell motility, cell division and cell and tissue morphogenesis. PMID- 19997134 TI - [Mental health among teenage boys remanded to prisoner]. AB - BACKGROUND: Incarceration of young people should be avoided according to Norwegian policy. Even so, young people reside in our prisons at all times. Our aim was to investigate their mental health. MATERIAL AND METHODS: Among 42 teenage boys (below 20 years of age) remanded to prison, 40 consented to participate in the study. Socio-demographic data were collected and a structured diagnostic instrument (K-SADS) was used to assess their mental health. RESULTS: Only four participants had no mental disorder. The 36 who had mental disorders had a mean of 2.5 diagnoses. The most common diagnosis was substance abuse/dependence (n = 31). 30 of the adolescents had serious conduct disorders and nine of them fulfilled the ICD-10 criteria for a dissocial personality disorder. Anxiety and/or depressive disorders (n = 14) and hyperkinetic disorders (n = 12) were also frequent. Many of the adolescents had unmet treatment needs in addition to large social problems, such as disrupted education and unemployment. Three of four adolescents had an immigrant background. INTERPRETATION: Teenage boys remanded to prison have a variety of complex problems. They have an extensive need for psychiatric treatment, drug treatment, education and employment. This calls for multidisciplinary coordinated interventions to avoid further negative development of chronic maladjustment with substance abuse, mental problems and crime. PMID- 19997135 TI - [Lesbian women's health-promoting experiences]. AB - BACKGROUND: For lesbian women, we know more about causes of health problems than health-promoting factors. The aim of this study was to explore what lesbian women have perceived as health-promoting experiences. MATERIAL AND METHODS: Focus group study with two group interviews (with nine lesbian women aged 40-55 years). Participants were required to be "out of the closet" regarding sexual orientation and to feel comfortable with their lives as lesbians. The interviews were audiotaped, transcribed, and analyzed with systematic text condensation supported by a salutogenic frame of reference. RESULTS: The women told about how a perception of being different could translate into opportunities and a positive strength. A good "coming-out process" could lead to companionship and strategies for coping with challenges. Furthermore, feelings of exemption from expectations of a narrow gender role had a positive impact on identity and sexuality. Finally, the liberating consequences of being able to organize personal relationships and family according to own priorities opened up new possibilities, especially regarding choosing to have children or not. INTERPRETATION: Health promoting strategies and initiatives must be grounded in what individuals consider to be important in their own lives. Knowledge about lesbians' health-promoting experiences may contribute to a positive focus on coping with minority stress and challenge an established understanding of pathology in a marginalized group. PMID- 19997136 TI - [Simple electrophysiologic examination of patients with palpitation or tachycardia]. AB - BACKGROUND: Discomfort because of palpitation and tachycardia is a frequent phenomenon. Clinical practice for diagnosing tachycardia has a low sensitivity, and patients without a correct diagnosis are often not treated optimally. MATERIAL AND METHODS: Results from all 24-hour-ECGs taken at Telemark Hospital in the period 1.8.04-31.7.05 were reviewed. Indications and findings were retrieved from the hospital database. 252 of totally 614 ECGs were performed for the indications palpitation and tachycardia. 137 patients with an unclear diagnosis were offered a simple electrophysiologic examination with stimulation of the right atrium by a pacemaker electrode. 38 of these patients were included and underwent the examination. An additional 18 patients were referred to a simple electrophysiologic examination based on the same criteria. RESULTS: 24-hour-ECG provided a diagnosis for 29 patients with palpitation and tachycardia. A simple electrophysiologic examination uncovered reentry tachycardia in nine patients and paroxystic atrial flutter in three patients. A total of 56 electrophysiologic examinations were performed, uncovering 12 cases of tachycardia suitable for ablation. Two patients had to undergo DC-conversion, no other complication was observed. INTERPRETATION: A simplified electrophysiologic examination of this group of patients will uncover tachycardia suitable for ablation in approximately 20 % of the patients, and increase diagnostic yield of supraventricular tachycardia by 64 %. PMID- 19997137 TI - [Immune reconstitution inflammatory syndrome and treatment of HIV infection]. AB - BACKGROUND: HAART (highly active antiretroviral therapy) may trigger a condition known as IRIS (immune reconstitution inflammatory syndrome); i.e. a paradoxical reaction to latent infections associated with reconstitution of the immune system. The article provides an overview of the syndrome and discusses diagnosis, risk factors and management. MATERIAL AND METHODS: The basis for the article was literature identified through non-systematic searches in PubMed and clinical experience. RESULTS: IRIS typically occurs some weeks to months after initiation of HAART, usually in association with mycobacterial infections, cytomegalovirus, Cryptococcus neoformans and Pneumocystis jirovecii. In principle, any pathogen may cause a similar inflammatory response. Risk factors for IRIS include severe immunodeficiency, high antigen burden and rapid immune response to HAART. The prognosis is good. However, treatment of infections must not delay the initiation of HAART, as such a delay may increase morbidity and mortality. HAART should be continued unless symptoms are life-threatening or likely to cause permanent sequelae. Corticosteroids may be helpful in cases with lesions in the central nervous system, obstructive lymph nodes or increasing respiratory symptoms. INTERPRETATION: Treatment of HIV infection has improved substantially, which implies an increased number of patients developing IRIS. A quick diagnosis and correct and timely treatment of opportunistic infections is important for the prognosis. PMID- 19997138 TI - [Calcium antagonists in treatment of hypertension]. AB - A sizeable number of antihypertensive drugs from different drug classes have received market authorization. New results from clinical studies are steadily launched, and this is also the case for calcium antagonists, which were established as antihypertensive drugs already in the 1970s. Which place do calcium antagonists have in the present treatment of hypertension? PMID- 19997139 TI - [Diagnostic challenges in a man with subarachnoidal haemorrhage]. AB - A 39-year-old man with multiple sclerosis was admitted to the Neurological department because of sudden and intense headache, photophobia, nausea and vomiting. A subarachnoidal haemorrhage was suspected and cerebral CT scanning showed small amounts of subarachnoidal blood distributed symmetrically over both cerebral hemispheres - an unusual location for aneurismal bleeding. Liquor analysis revealed erythrocytes, positive bilirubin and slightly elevated protein. Subsequent cerebral MRI showed no signs of aneurysm, vascular malformation, venous sinus thrombosis or infarction. An intraarterial cerebral angiography was then performed and showed bilateral widespread segmental vasoconstriction in the anterior, middle and posterior cerebral arteries. A diagnostic workup on systemic vasculitis was negative. The patient's good general condition in spite of extensive angiographic findings rendered the diagnosis cerebral vasculitis unlikely. A state of reversible cerebral segmental vasoconstriction was suspected and the patient was treated with the calcium antagonist nifedipine and observed for four weeks. Cerebral angiography was then repeated and found to be normal. This case report highlights a less recognized and probably underdiagnosed cause of subarachnoidal bleeding. PMID- 19997140 TI - [Just a shadow of reality]. PMID- 19997141 TI - [Diabetes in pregnancy--prevention over generations]. PMID- 19997142 TI - [Extreme weight reduction without surgery]. PMID- 19997143 TI - [Patients on maintenance therapy for opiate dependence should be treated for hepatitis C]. PMID- 19997144 TI - [Short-time hepatitis C treatment and drug abuse]. PMID- 19997145 TI - [Hepatitis C is important for drug addicts!]. PMID- 19997146 TI - [Is the reader manipulated?]. PMID- 19997148 TI - [Epidemics and danger of infection]. PMID- 19997149 TI - [Consent and information]. PMID- 19997151 TI - [How to combat HIV, tuberculosis and malaria?]. AB - BACKGROUND: In year 2 000, the United Nations (UN) agreed on eight millennium development goals (MDGs). Goal number 6 is to combat HIV/AIDS, malaria and other communicable diseases, including tuberculosis. The aim of this paper is to provide an overview of current status and prognosis for this MDG, and to discuss strategies that need to be implemented to reach the goal. MATERIAL AND METHODS: The article is mainly based on publications from the UN or WHO-affiliated organizations. RESULTS: The global HIV prevalence rate has remained at 0.8 % for the last ten years, but the total infected population is still increasing. Access to treatment has increased considerably, but only 28 % of those in need of treatment (and living in developing countries), received it in 2007. Global tuberculosis incidence and prevalence rates of tuberculosis are falling, but not in Europe and Africa. For malaria, there is more uncertainty, but there seems to be a declining incidence in many countries. INTERPRETATION: The MDG for HIV/AIDS cannot be reached by treatment alone, continued emphasis on prevention and more specific prevention strategies is necessary. The global targets for tuberculosis can be reached, but probably not in Africa; in addition, multi-resistant tuberculosis is increasing. Modern combination treatment, impregnated bednets and indoor residual spraying has led to a substantial reduction in the prevalence of malaria during few years in African countries. PMID- 19997152 TI - [New approach to resistant tuberculosis]. PMID- 19997153 TI - Photorealistic images of carpet cloaks. AB - Using home-built dedicated ray-tracing software, we simulate photorealistic images of sceneries in three dimensions including dielectric carpet cloaks--i.e., continuously varying refractive-index distributions that allow for invisibility cloaking of a bump in a metallic carpet. Results for the ideal and for a simplified cloak are shown. The presented material gives a visual and intuitive impression of the performance of different arrangements and might be ideally suited for communicating the concepts of transformation optics to the general public. PMID- 19997154 TI - Optical polarization based logic functions (XOR or XNOR) with nonlinear Gallium nitride nanoslab. AB - We present a scheme of XOR/XNOR logic gate, based on non phase-matched noncollinear second harmonic generation from a medium of suitable crystalline symmetry, Gallium nitride. The polarization of the noncollinear generated beam is a function of the polarization of both pump beams, thus we experimentally investigated all possible polarization combinations, evidencing that only some of them are allowed and that the nonlinear interaction of optical signals behaves as a polarization based XOR. The experimental results show the peculiarity of the nonlinear optical response associated with noncollinear excitation, and are explained using the expression for the effective second order optical nonlinearity in noncollinear scheme. PMID- 19997155 TI - Ultrabroad emission from a bismuth doped chalcogenide glass. AB - We report emission from a bismuth doped chalcogenide glass which is flattened, has a full width at half maximum (FWHM) of 600 nm, peaks at 1300 nm and covers the entire telecommunications window. At cryogenic temperatures the FWHM reaches 850 nm. The quantum efficiency and lifetime were as high as 32% and 175 mus, respectively. We also report two new bismuth emission bands at 2000 and 2600 nm. Absorption bands at 680, 850, 1020 and 1180 nm were observed. The 1180 nm absorption band was previously unobserved. We suggest that the origin of the emission in Bi:GLS is Bi22- dimers. PMID- 19997156 TI - Thermal tunability of photonic bandgaps in liquid crystal infiltrated microstructured polymer optical fibers. AB - We demonstrate the photonic bandgap effect and the thermal tunability of bandgaps in microstructured polymer optical fibers infiltrated with liquid crystal. Two liquid crystals with opposite sign of the temperature gradient of the ordinary refractive index (E7 and MDA-00-1444) are used to demonstrate that both signs of the thermal tunability of the bandgaps are possible. The useful bandgaps are ultimately bounded to the visible range by the transparency window of the polymer. PMID- 19997157 TI - Airy beams generated by a binary phase element made of polymer-dispersed liquid crystals. AB - Using polymer-dispersed liquid crystals (PDLCs), an electrically switchable binary phase pattern was fabricated to generate Airy beams through a programmable lithographic system. The right main lobe of the reconstructed Airy beam experienced 1.3 mm transverse deflection within 24 cm propagation distance. With a suitable voltage applied, the binary PDLC pattern can be erased due to the index match between polymers and liquid crystals. This versatile approach can be also used to generate other special beams with electrically tunable capability. PMID- 19997158 TI - Optical absorption enhancement in silicon nanowire arrays with a large lattice constant for photovoltaic applications. AB - In this paper, we use the transfer matrix method to calculate the optical absorptance of vertically-aligned silicon nanowire (SiNW) arrays. For fixed filling ratio, significant optical absorption enhancement occurs when the lattice constant is increased from 100 nm to 600 nm. The enhancement arises from an increase in field concentration within the nanowire as well as excitation of guided resonance modes. We quantify the absorption enhancement in terms of ultimate efficiency. Results show that an optimized SiNW array with lattice constant of 600 nm and wire diameter of 540 nm has a 72.4% higher ultimate efficiency than a Si thin film of equal thickness. The enhancement effect can be maintained over a large range of incidence angles. PMID- 19997160 TI - Coupling of gap plasmons in multi-wire waveguides. AB - We investigate the coupling of gap plasmons in various configurations of neighboring metallic nanowires. Starting with the basic element defining a gap plasmon, consisting of two neighboring silver wires, we study the energy splitting and symmetry properties of hybridized plasmons resulting from the interaction of two wire pairs. The system is shown to display non-avoided crossings of hybridized modes, and it evolves at short distances towards a degenerate system consisting of four wires arranged in a square, where two new gap plasmons emerge from redshifted higher-energy modes. The gap modes of three neighboring wires are also described in a continuous transition from a coplanar configuration to an equilateral triangle arrangement. The interaction between wire pairs is shown to be weak enough to prevent efficient transfer of plasmon signal from a pair to the other one, which is beneficial to avoid crosstalking, but not to produce waveguide couplers. The coupling is significantly increased by placing a wire of rectangular cross section in between the wire pairs, thus allowing us to achieve large plasmon-signal transfers within propagation distances below the attenuation length. Our results can find application in the design of signal-processing devices based upon gap plasmons. PMID- 19997159 TI - Adaptive optics optical coherence tomography at 120,000 depth scans/s for non invasive cellular phenotyping of the living human retina. AB - This paper presents a successful combination of ultra-high speed (120,000 depth scans/s), ultra-high resolution optical coherence tomography with adaptive optics and an achromatizing lens for compensation of monochromatic and longitudinal chromatic ocular aberrations, respectively, allowing for non-invasive volumetric imaging in normal and pathologic human retinas at cellular resolution. The capability of this imaging system is demonstrated here through preliminary studies by probing cellular intraretinal structures that have not been accessible so far with in vivo, non-invasive, label-free imaging techniques, including pigment epithelial cells, micro-vasculature of the choriocapillaris, single nerve fibre bundles and collagenous plates of the lamina cribrosa in the optic nerve head. In addition, the volumetric extent of cone loss in two colour-blinds could be quantified for the first time. This novel technique provides opportunities to enhance the understanding of retinal pathogenesis and early diagnosis of retinal diseases. PMID- 19997161 TI - Optical mirror trap with a large field of view. AB - Holographic optical tweezers typically require microscope objectives with high numerical aperture and thus usually suffer from the disadvantage of a small field of view and a small working distance. We experimentally investigate an optical mirror trap that is created after reflection of two holographically shaped collinear beams on a mirror. This approach combines a large field of view and a large working distance with the possibility to manipulate particles in a large size range, since it allows to use a microscope objective with a numerical aperture as low as 0.2. In this work we demonstrate robust optical three dimensional trapping in a range of 1mm x 1mm x 2mm with particle sizes ranging from 1.4 mum up to 45 mum. The use of spatial light modulator based holographic methods to create the trapping beams allows to simultaneously trap many beads in complex, dynamic configurations. We present measurements that characterize the mirror traps in terms of trap stiffness, maximum trapping force and capture range. PMID- 19997162 TI - Freezing the carrier-envelope phase of few-cycle light pulses about a focus. AB - We study the effects of dispersive media and structures on the carrier-envelope phase (CEP) shift of focused few-cycles pulses. For phase-sensitive interactions with matter requiring focusing in vacuum, the variation of the CEP through the focal region can be significantly slow down by inserting a dispersive slab of adequate thickness between the focusing system and the focus. The focal CEP shift can also be slow down in experiments requiring focusing in a dispersive medium by a suitable choice of the dispersive propagation distance up to the focus. PMID- 19997163 TI - Improved bending loss characteristics of asymmetric surface plasmonic waveguides for flexible optical wiring. AB - We present improved characteristics of the curved plasmonic waveguide which consists of a thin metal stripe with asymmetric cladding layers. It is shown that in the proposed curved asymmetric plasmonic waveguides, a balance between a radiation due to bending and a radiation due to the asymmetric claddings allows a bending with a smaller radius curvature and a lower loss compared to the waveguide with symmetric claddings. At the same time, a symmetric metal stripe waveguide's typical trade-off between the bending characteristics and the propagation loss of a straight waveguide is relaxed with proper amount of asymmetry. With the proposed structure, a plasmonic waveguide bending whose radius is as small as 2 mm with a total loss of 1.8 dB/90 degrees is designed. Enhanced sensitivity to the surrounding medium and its application are discussed. PMID- 19997164 TI - Observation of electromagnetically induced transparency and slow light in the dark state--bright state basis. AB - The quantum coherence phenomenon of electromagnetically induced transparency (EIT) is observed in a three-level system composed of an excited state and two coherent superpositions of the two ground-state levels. This peculiar ground state basis is composed of the so-called bright and dark states of the same atomic system in a standard coherent population trapping configuration. The characteristics of EIT, namely, width of the transmission window and reduced group velocity of light, in this unusual basis, are theoretically and experimentally investigated and are shown to be essentially identical to those of standard EIT in the same system. PMID- 19997165 TI - Optical singularities associated with the energy flow of two closely spaced core shell nanocylinders. AB - Optical singularities associated with the energy flow of two closely spaced dielectric-core gold-shell nanocylinders are studied by two-dimensional finite difference time domain method. The simulation results show that optical vortices as well as saddle points can be observed in the energy flow pattern of light interacting with the core-shell nanocylinder pair in its in-phase symmetric dipolar plasmon mode. The rotating direction of the optical vortices can be tuned by varying the width of the gap between the nanocylinder pair and the value of the permittivity of the dielectric core. PMID- 19997166 TI - Tunable optical metamaterial based on liquid crystal-gold nanosphere composite. AB - Effect of the surrounding anisotropic liquid crystal medium on the surface plasmon resonance (SPR) exhibited by concentrated suspensions of gold nanospheres has been investigated experimentally and compared with the Mie scattering theory. The observed polarization-sensitive SPR and the red-shift in the SPR wavelength with increasing concentration of the gold nanospheres in the liquid crystal matrix have been explained using calculations based on the Maxwell Garnet effective medium theory. Agglomeration of the gold nanospheres that could also lead to such a red-shift has been ruled out using Atomic force microscopy study of thin nanoparticle-doped smectic films obtained on solid substrates. Our study demonstrates feasibility of obtaining tunable optical bulk metamaterials based on smectic liquid crystal - nanoparticle composites. PMID- 19997167 TI - Optical fiber nanoprobe preparation for near-field optical microscopy by chemical etching under surface tension and capillary action. AB - We propose a technique of chemical etching for fabrication of near perfect optical fiber nanoprobe (NNP). It uses photosensitive single mode optical fiber to etch in hydro fluoric (HF) acid solution. The difference in etching rate for cladding and photosensitive core in HF acid solution creates capillary ring along core-cladding boundary under a given condition. The capillary ring is filled with acid solution due to surface tension and capillary action. Finally it creates near perfect symmetric tip at the apex of the fiber as the height of the acid level in capillary ring decreases while width of the ring increases with continuous etching. Typical tip features are short taper length (approximately 4 microm), large cone angle (approximately 38 degrees ), and small probe tip dimension (<100 nm). A finite difference time domain (FDTD) analysis is also presented to compare near field optics of the NNP with conventional nanoprobe (CNP). The probe may be ideal for near field optical imaging and sensor applications. PMID- 19997168 TI - Plasmonic nano lithography with a high scan speed contact probe. AB - We demonstrate plasmonic lithography with an optical contact probe to achieve high speed patterning without external gap distance control between the probe and the photoresist. The bottom surface of the probe is covered with a 10 nm thickness silica glass film for the gap distance control and coated with self assembled monolayer (SAM) to reduce friction between the probe and the photoresist. We achieve a patterning resolution of ~50 nm and a patterning speed of ~10 mm/s. We obtain the quality of line patterning comparable to that in conventional optical lithography. PMID- 19997169 TI - Simultaneous dual-band optical coherence tomography in the spectral domain for high resolution in vivo imaging. AB - Optical coherence tomography (OCT) in the spectral domain is demonstrated simultaneously at two wavelength bands centered at 800 nm and 1250 nm. A novel commercial supercontinuum laser is applied as a single low coherence broadband light source. The emission spectrum of the source is shaped by optical and spatial filtering in order to achieve an adequate double peak spectrum containing the wavelength bands 700 - 900 nm and 1100 - 1400 nm for dual-band OCT imaging and thus reducing the radiation exposure of the sample. Each wavelength band is analyzed with an individual spectrometer at an A-scan rate of about 12 kHz which enables real-time imaging for the examination of moving samples. A common path optical setup optimized for both spectral regions with a separate single fiber based scanning unit was realized which facilitates flexible handling and easy access to the measurement area. The free-space axial resolutions were measured to be less than 4.5 microm and 7 microm at 800 nm and 1250 nm, respectively. Three dimensional imaging ten times faster than previously reported with a signal-to noise-ratio of above 90 dB is achieved simultaneously in both wavelength bands. Spectral domain dual-band OCT combines real-time imaging with high resolution at 800 nm and enhanced penetration depth at 1250 nm and therefore provides a well suited tool for in vivo vasodynamic measurements. Further, spatially resolved spectral features of the sample are obtained by means of comparing the backscattering properties at two different wavelength bands. The ability of dual band OCT to enhance tissue contrast and the sensitivity of this imaging modality to wavelength-dependent sample birefringence is demonstrated. PMID- 19997170 TI - Full duplex 60-GHz RoF link employing tandem single sideband modulation scheme and high spectral efficiency modulation format. AB - This study proposes a full duplex 60-GHz band radio-over-fiber (RoF) link using a modified tandem single sideband (TSSB) modulation scheme with frequency doubling. Based on the modified TSSB modulation scheme, no dispersion induced fading is observed; high spectral efficiency vector signal can be utilized; and wavelength reuse can also be achieved. Both single carrier 8-QAM and QPSK-OFDM signals for down-link transmissions are experimentally demonstrated. After transmission of 50 km SSMF, no significant receiver power penalties are observed. Wavelength reuses with 1.25-Gb/s OOK using a reflective semiconductor optical amplifier (RSOA) for up-link transmission are also demonstrated. After transmission of 50-km SSMF, no significant receiver power penalties are also observed. PMID- 19997171 TI - Model for passive coherent beam combining in fiber laser arrays. AB - We present a new model for studying the beam combining mechanism, spectral and temporal dynamics, the role of nonlinearity, and the power scaling issue of discretely coupled fiber laser arrays. The model accounts for the multiple longitudinal modes of individual fiber lasers and shows directly the formation of the composite-cavity modes. Detailed output power spectra and their evolution with increasing array size and pump power are also explored for the first time. In addition, it is, to our knowledge, the only model that closely resembles the real experimental conditions in which no deliberate control of the fiber lengths (mismatch) is required while highly efficient coherent beam combining is still attained. PMID- 19997172 TI - RF magnetron sputtered (BiDy)3(FeGa)5O12:Bi2O3 composite garnet-oxide materials possessing record magneto-optic quality in the visible spectral region. AB - Bismuth-substituted iron garnets are considered to be the most promising magneto optical materials because of their excellent optical transparency and very high magneto-optical figures of merit in the near-infrared spectral region. However, the practical application of garnets in the visible and short-wavelength infrared parts of spectrum is currently limited, due to their very high optical absorption (especially in sputtered films) in these spectral regions. In this paper, we identify the likely source of excess absorption observed in sputtered garnet films in comparison with epitaxial layers and demonstrate (Bi,Dy)(3)(Fe,Ga)(5)O(12): Bi(2)O(3) composites possessing record MO quality in the visible region. PMID- 19997173 TI - Synchronization of bandwidth-enhanced chaos in semiconductor lasers with optical feedback and injection. AB - We experimentally investigate the generation and synchronization of bandwidth enhanced chaos in a semiconductor laser (drive laser) that is subject to optical injection from another chaotic semiconductor laser (injection laser) with optical feedback. Effective bandwidth enhancement is achieved over 12 GHz, under the condition in which the optical wavelength of the drive laser is positively detuned with respect to that of the injection laser, outside the injection locking range. The bandwidth-enhanced chaotic signal of the drive laser is injected into a third semiconductor laser (response laser) for synchronization. Synchronization of chaos with a bandwidth greater than 12 GHz is observed between the drive and response lasers, under the condition in which the optical wavelength of the response laser is negatively detuned with respect to that of the drive laser, satisfying the injection locking condition. High-quality chaos synchronization is observed within the injection locking range between the drive and response lasers and under the condition of a low relaxation oscillation frequency of the response laser. PMID- 19997174 TI - Dual polarization fiber grating laser hydrophone. AB - A novel fiber optic hydrophone based on the integration of a dual polarization fiber grating laser and an elastic diaphragm is proposed and experimentally demonstrated. The diaphragm transforms the acoustic pressure into transversal force acting on the laser cavity which changes the fiber birefringence and therefore the beat frequency between the two polarization lines. The proposed hydrophone has advantages of ease of interrogation, absolute frequency encoding, and capability to multiplex a number of sensors on a single fiber by use of frequency division multiplexing technique. PMID- 19997175 TI - Phasing rectangular apertures. AB - Several techniques have been developed to phase apertures in the context of astronomical telescopes with segmented mirrors. Phasing multiple apertures, however, is important in a wide range of optical applications. The application of primary interest in this paper is the phasing of multiple short pulse laser beams for fast ignition fusion experiments. In this paper analytic expressions are derived for parameters such as the far-field distribution, a line-integrated form of the far-field distribution that could be fit to measured data, enclosed energy or energy-in-a-bucket and center-of-mass that can then be used to phase two rectangular apertures. Experimental data is taken with a MEMS device to simulate the two apertures and comparisons are made between the analytic parameters and those derived from the measurements. Two methods, fitting the measured far-field distribution to the theoretical distribution and measuring the ensquared energy in the far-field, produced overall phase variance between the 100 measurements of less than 0.005 rad(2) or an RMS displacement of less than 12 nm. PMID- 19997176 TI - Time-gated perturbation Monte Carlo for whole body functional imaging in small animals. AB - This paper explores a time-resolved functional imaging method based on Monte Carlo model for whole-body functional imaging of small animals. To improve the spatial resolution and quantitative accuracy of the functional map, a Bayesian hierarchical method with a high resolution spatial prior is applied to guide the optical reconstructions. Simulated data using the proposed approach are employed on an anatomically accurate mouse model where the optical properties range and volume limitations of the diffusion equation model exist. We investigate the performances of using time-gated data type and spatial priors to quantitatively image the functional parameters of multiple organs. Accurate reconstructions of the two main functional parameters of the blood volume and the relative oxygenation are demonstrated by using our method. Moreover, nonlinear optode settings guided by anatomical prior is proved to be critical to imaging small organs such as the heart. PMID- 19997177 TI - Scattering by solutions of major sea salts. AB - Increased scattering by seawater relative to that by pure water is primarily due to additional fluctuation of the refractive index contributed by sea salts. Salts with different ionic weight and sizes, while barely affecting the scattering that is due to density fluctuations, have a significant effect on the scattering that is due to concentration fluctuations. And this explains the major differences of their total scattering that would be observed. Scattering by solutions of NaCl, the major sea salt, is consistently about 6.7% and 4% lower than seawater of the same mass concentration and of the same refractive index, respectively. Because of ionic interactions, the molecular scattering does not follow the simple addition rule that applies to bulk inherent optical properties, with the total less than the summation of the parts. The possible values of scattering by waters of, such as, Dead Sea or Orca Basin, which have different salt composition from seawater, are discussed. PMID- 19997178 TI - Guided resonances in photonic crystals with point-defected aperiodically-ordered supercells. AB - In this paper, we study the excitation of guided resonances (GRs) in photonic crystal slabs based on point-defected aperiodically-ordered supercells. With specific reference to perforated-slab structures and the Ammann-Beenker octagonal lattice geometry, we carry out full-wave numerical studies of the plane-wave responses and of the underlying modal structures, which illustrate the representative effects induced by the introduction of symmetry-preserving and symmetry-breaking defects. Our results demonstrate that breaking the supercell mirror symmetries via the judicious introduction of point-defects enables for the excitation of otherwise uncoupled GRs, with control on the symmetry properties of their field distributions, thereby constituting an attractive alternative to those GR-engineering approaches based on the asymmetrization of the hole shape. In this framework, aperiodically-ordered supercells seem to be inherently suited, in view of the variety of inequivalent defect sites that they can offer. PMID- 19997179 TI - Stripe-array diode-laser in an off-axis external cavity: theory and experiment. AB - Stripe-array diode lasers naturally operate in an anti-phase supermode. This produces a sharp double lobe far field at angles +/-alpha depending on the period of the array. In this paper a 40 emitter gain guided stripe-array laterally coupled by off-axis filtered feedback is investigated experimentally and numerically. We predict theoretically and confirm experimentally that at doubled feedback angle 2alpha a stable higher order supermode exists with twice the number of emitters per array period. The theoretical model is based on time domain traveling wave equations for optical fields coupled to the carrier density equation taking into account diffusion of carriers. Feedback from the external reflector is modeled using Fresnel integration. PMID- 19997180 TI - Enhanced photo-assisted electrical gating in vanadium dioxide based on saturation induced gain modulation of erbium-doped fiber amplifier. AB - By incorporating saturation-induced gain modulation of an erbium-doped fiber amplifier (EDFA), we have demonstrated a high-speed photo-assisted electrical gating with considerably enhanced switching characteristics in a two-terminal device fabricated by using vanadium dioxide thin film. The gating operation was performed by illuminating the output light of the EDFA, whose transient gain was modulated by adjusting the chopping frequency of the input light down to 1 kHz, onto the device. In the proposed gating scheme, gated signals with a temporal duration of approximately 40 micros were successively generated at a repetition rate of 1 kHz. PMID- 19997181 TI - A linearly-polarized Nd:YVO4/KTP microchip green laser. AB - We described the principle and the fabrication of a Nd:YVO(4)/KTP microchip for the linearly-polarized green laser and verified its availability by manufacturing and characterizing the green laser using the microchip. Under the driving condition having the modulation frequency of 60 Hz and the duty ratio of 25%, the laser showed the stable linear polarization, the maximum average power of 37 mW, yielding the high electrical-to-optical efficiency of 10.9%. PMID- 19997182 TI - Femtosecond two-photon absorption measurements based on the accumulative photo thermal effect and the Rayleigh interferometer. AB - A rapid, straightforward method for measuring the two-photon absorption cross sections in liquid samples based on both the accumulative photo-thermal effect and the Rayleigh interferometery is described and demonstrated. This technique combines the sensitivity of the thermal lens approach and the accuracy of interferometry techniques. Focusing a high repetition rate laser beam in the sample, generating a localized change in its refractive index, induces the photo thermal phase shift. By recording and processing two interference patterns, this technique allows the rapid estimation of the two-photon absorption cross section of the sample. Significantly, the experimental results demonstrate that this new method can be used with both fluorescent and non-fluorescent samples. PMID- 19997183 TI - Modes of shallow photonic crystal waveguides: semi-analytic treatment. AB - We investigate the formation of photonic crystal waveguide (PCW) modes within the framework of perturbation theory. We derive a differential equation governing the envelope of PCW modes constructed from weak perturbations using an effective mass formulation based on the Luttinger-Kohn method from solid-state physics. The solution of this equation gives the frequency of the mode and its field. The differential equation lends itself to simple analytic approximations which reduce the problem to that of solving slab waveguide modes. By using this model, we demonstrate that the nature of the projected band structure and corresponding Bloch functions are central to the behaviour of PCW modes. With this understanding, we explain why the odd mode in a hexagonal PCW spans the entire Brillouin zone while the even mode is cut off. PMID- 19997184 TI - Polarization sensitive, three-dimensional, single-molecule imaging of cells with a double-helix system. AB - Double-helix point spread function photoactivation-localization microscopy allows three-dimensional (3D) superresolution imaging of objects smaller than the optical diffraction-limit. We demonstrate polarization sensitive detection with 3D super-localization of single-molecules and unveil 3D polarization specific characteristics of single-molecules within the intracellular structure of PtK1 cells expressing photoactivatable green fluorescent protein. The system modulates orthogonal polarization components of single-molecule emissions with a single spatial light modulator and detects them separately with a single detector. Information obtained from the two polarization channels demonstrates polarization based contrast in 3D superresolution imaging. Further, we show that the 3D information from the two channels can be optimally combined to yield up to 30% improvement in localization precision relative to a single polarization channel system. PMID- 19997185 TI - Metamaterial electromagnetic concentrators with arbitrary geometries. AB - The electromagnetic concentrators play an important role in the harnessing of light in solar cells or similar devices, where high field intensities are required. The material parameters for two-dimensional (2D) metamaterial-assisted electromagnetic concentrators with arbitrary geometries are derived based on transformation-optical approach. Enhancements in field intensities of the 2D concentrator have been shown by full-wave simulation. All theoretical and numerical results validate the material parameters for the 2D concentrator with irregular cross section we developed. PMID- 19997186 TI - Band-limited angular spectrum method for numerical simulation of free-space propagation in far and near fields. AB - A novel method is proposed for simulating free-space propagation. This method is an improvement of the angular spectrum method (AS). The AS does not include any approximation of the propagation distance, because the formula thereof is derived directly from the Rayleigh-Sommerfeld equation. However, the AS is not an all round method, because it produces severe numerical errors due to a sampling problem of the transfer function even in Fresnel regions. The proposed method resolves this problem by limiting the bandwidth of the propagation field and also expands the region in which exact fields can be calculated by the AS. A discussion on the validity of limiting the bandwidth is also presented. PMID- 19997187 TI - Assessing light scattering of intracellular organelles in single intact living cells. AB - This report presents a model-independent method of assessing contributions to the light scattering from individual organelles in single intact cells. We first measure the 3D index map of a living cell, and then modify the map in such a way so as to eliminate contrast due to a particular intracellular organelle. By calculating and comparing the light scattering distributions calculated from the original and modified index maps using the Rytov approximation, we extract the light scattering contribution from the particular organelle of interest. The relative contributions of the nucleus and nucleolus to the scattering of the entire cell are thus determined, and the applicability of the homogeneous spherical model to non-spherical and heterogeneous organelles in forward scattering is evaluated. PMID- 19997188 TI - Interaction-induced Lipkin-Meshkov-Glick model in a Bose-Einstein condensate inside an optical cavity. AB - In this paper we present an experimentally feasible scheme to simulate a generalized Lipkin-Meskov-Glick model in a Bose-Einstein condensate coupled dispersively with an ultrahigh-finesse optical cavity. This obtained Hamiltonian has a unique advantage in that all parameters can be controlled independently by using Feshbach resonance technique, a pump laser along cavity axis and an external driving laser. By the proper choice of parameters, the macroscopic quantum coherent effect with a large amplitude can be successfully achieved. Comparing with the exist schemes, our proposal has a cleaner, perhaps significantly improved to observe this whole coherent effect. Finally, we predict a novel interaction-induced topological transition, which is an abrupt variation from pi to zero of the Berry phase. PMID- 19997189 TI - One-unit system to reconstruct a 3-D movie at a video-rate via electroholography. AB - We have developed a one-unit system, including creating and displaying a hologram for real-time reproduction of a three-dimensional image via electroholography. We have constructed this one-unit system by connecting a special-purpose computer for holography and a special display board with a reflective liquid crystal display as a spatial light modulator. Using this one-unit system, we succeeded in reproducing a three-dimensional image composed of 10,000 points at a speed of 30 frames per second, which is the video rate in NTSC format. In addition, we were able to control a three-dimensional image in real-time using our system. PMID- 19997190 TI - Transverse motion as a source of noise and reduced correlation of the Doppler phase shift in spectral domain OCT. AB - Recently, a new phase-resolved Doppler model was presented for spectral domain optical coherence tomography (SD OCT) showing that the linear relation between the axial velocity component of the obliquely moving sample and the phase difference of consecutive A-Scans does not hold true in the presence of a transverse velocity component which is neglected in the widely-used classic Doppler analysis. Besides taking note of the new non-proportional relationship of phase shift and oblique sample motion, it is essential to consider the correlation of the phase shift and its specific characteristic at certain Doppler angles for designing Doppler experiments with SD OCT. A correlation quotient is introduced to quantify the correlation of the backscattering signal in consecutive A-Scans as a function of the oblique sample motion. It was found that at certain velocities and Doppler angles no correlation of the phases of sequential A-Scans exists, even though the signal does not vanish. To indicate how the noise of the Doppler phase shift behaves for oblique movement, the standard deviation is determined as a function of the correlation quotient and the number of complex Doppler data averaged. The detailed theoretical model is validated by using a flow phantom model consisting of a 1% Intralipid flow through a 310 microm capillary. Finally, a short discussion of the presented results and the consequence for performing Doppler experiments is given. PMID- 19997191 TI - Collimated light from a waveguide for a display backlight. AB - We report light collimation from a point source without the space normally needed for fan-out. Rays emerge uniformly from all parts of the surface of a blunt wedge light-guide when a point source of light is placed at the thin end and the source's position determines ray direction in the manner of a lens. A lenticular array between this light-guide and a liquid crystal panel guides light from color light-emitting diodes to designated sub-pixels thereby removing the need for color filters and halving power consumption but we foresee much greater power economies and wider application. PMID- 19997192 TI - Three-photon N00N states generated by photon subtraction from double photon pairs. AB - We describe an experimental demonstration of a novel three-photon N00N state generation scheme using a single source of photons based on spontaneous parametric down-conversion (SPDC). The three-photon entangled state is generated when a photon is subtracted from a double pair of photons and detected by a heralding counter. Interference fringes measured with an emulated three-photon detector reveal the three-photon de Broglie wavelength and exhibit visibility > 70% without background subtraction. PMID- 19997193 TI - First experimental demonstration of 6Gb/s real-time optical OFDM transceivers incorporating channel estimation and variable power loading. AB - The fastest ever 6Gb/s real-time FPGA-based optical orthogonal frequency division multiplexing (OOFDM) transceivers utilizing channel estimation are experimentally demonstrated, for the first time, with variable power loading being incorporated to effectively compensate for the rapid system frequency response roll-off effect. The implemented transceivers are constructed entirely from off-the-shelf components and incorporate crucial functionalities of on-line performance monitoring and live optimization of key parameters including signal clipping, subcarrier power and operating conditions of directly modulated DFB lasers (DMLs). Real-time end-to-end transmission of a 6Gb/s 16-QAM-encoded OOFDM signal over 300m OM1 multi-mode fiber with a power penalty of 0.5dB is successfully achieved in an intensity-modulation and direct-detection system employing a DML. PMID- 19997194 TI - Cavity design and characteristics of monolithic long-wavelength InAs/InP quantum dash passively mode-locked lasers. AB - By extending the net-gain modulation phasor approach to account for the discrete distribution of the gain and saturable absorber sections in the cavity, a convenient model is derived and experimentally verified for the cavity design of two-section passively mode-locked quantum dash (QDash) lasers. The new set of equations can be used to predict functional device layouts using the measured modal gain and loss characteristics as input. It is shown to be a valuable tool for realizing the cavity design of monolithic long-wavelength InAs/InP QDash passively mode-locked lasers. PMID- 19997195 TI - A continuously tunable and filterless optical millimeter-wave generation via frequency octupling. AB - This work proposes a cost-effective, continuously tunable and filterless optical millimeter-wave (MMW) signal generation employing frequency octupling. Optical MMW signals with 30-dB undesired sideband suppression ratios can be obtained. Since no optical filtering is required, the proposed system can be readily implemented in wavelength-division-multiplexing (WDM) systems. V-band 60-GHz and W-band 80-GHz optical MMW signals are experimentally demonstrated. Because of the high undesired sideband suppression ratio, 60-GHz waveform with 50% duty cycle is observed. The single-sideband (SSB) phase noise of the generated 60-GHz signal is -73 dBc/Hz at 10 kHz. The proposed system is a viable solution for the future ultra-high frequency MMW applications up to 320 GHz using the external modulator with a limited bandwidth of 40 GHz. PMID- 19997196 TI - An experimental study of the plasmonic Talbot effect. AB - The Talbot effect of the surface plasmon polaritons (SPPs) using SPP launching gratings is studied experimentally. Talbot carpets are obtained and the Talbot distance is given when the paraxial approximation is not satisfied. Multi-layer and multi-level-phase launching gratings are designed to enhance the intensities of the amplitude-modulated revivals. Effective focusing of SPPs with multiple focal spots and a subwavelength full width at half maximum is obtained by using a three-layer four-level-phase launching grating. PMID- 19997197 TI - Three distinct sarcomeric patterns of skeletal muscle revealed by SHG and TPEF microscopy. AB - We have extensively characterized the sarcomeric SHG signal as a function of animal species (rat versus xenopus), age (adult versus larval) and tissue preparation (fixed or fresh) and we found that the main feature of this signal is a single peak per mature sarcomere (about 85% of all sarcomeres). The remaining (15%) was found to be either double peak per mature sarcomere or mini sarcomeres (half of a sarcomere) using alpha-actinin immuno detection of the Z-band. The mini sarcomeres are often found in region of pitchfork-like SHG pattern. We suggest that double peak SHG pattern could indicate regions of sarcomeric proteolysis whereas pitchfork-like SHG pattern could reveal sarcomeric assembly. PMID- 19997198 TI - A novel phase measuring deflectometry for aspheric mirror test. AB - A method based on fringe reflection is presented to measure the aspheric mirror with higher precise. This method measures the absolute height of the aspheric mirror with dummy paraboloid, while the camera is located beside the optical axis of the test surface. It can be used to measure the aspheric mirror with high deviation. And for locating the camera beside the optical axis, this method doesn't have occlusion problem and can do the measurement more flexibility. Furthermore, compared with the traditional PMD, this method measures the tested surface with dummy paraboloid instead of the intersection of two straight lines, so it doesn't need to calibrate the image projection vectors. And the errors of the calibration influence this method less than the traditional method. Therefore, this method can measure the high deviation aspheric mirror with higher precise even if the calibration precise isn't very high. Computer simulations and preliminary experiment validate the feasibility of this method. PMID- 19997199 TI - Fiber Bragg gratings with enhanced thermal stability by residual stress relaxation. AB - Fiber Bragg gratings with greatly enhanced thermal stability have been fabricated by the use of femtosecond laser pulse irradiation on optical fibers with relaxed residual stress, through using high temperature annealing treatment. The grating reflectivity and resonant wavelength can be maintained for periods up to 20 hours using isothermal measurements and temperatures up to 1200 degrees C. No hysteresis was observed in the wavelength response when the gratings were annealed and the temperature cycled repeatedly between room temperature and 1200 degrees C. PMID- 19997200 TI - Omnidirectional constant transmission and negative Brewster angle at planar interfaces associated with a uniaxial medium. AB - This paper presents our detailedly theoretical analyses on omnidirectional constant transmission and negative Brewster angle at planar interfaces associated with a uniaxial medium. The amplitude reflection and transmission coefficients at planar interfaces associated with uniaxial media are derived by examining the boundary condition and the dispersion relation. It is found that under certain conditions, the coefficients are constants independent of the incident angle. Another interesting phenomenon is that an interface between isotropic and uniaxial media can exhibit negative refraction under Brewster condition. Our results offer considerable potential device applications of uniaxial media. PMID- 19997201 TI - Arbitrary-order all-fiber temporal differentiator based on a fiber Bragg grating: design and experimental demonstration. AB - A new technique to design an all-fiber temporal differentiator that has a large bandwidth and an arbitrary differentiation order is proposed and investigated. The proposed temporal differentiator is a special fiber Bragg grating (FBG) that is designed by controlling its magnitude and phase responses with the discrete layer peeling (DLP) method. There are three important features of this technique: 1) the temporal differentiator has an arbitrary magnitude response and a controllable bandwidth; 2) the temporal differentiator can be designed and fabricated with an arbitrary differentiation order that is realized in a single FBG; 3) the required maximum index modulation of the FBG-based differentiator is largely decreased by using a Gaussian windowing function. The use of the proposed technique to design temporal differentiators with a differentiation order up to the fourth and with a bandwidth up to 500 GHz is studied. A proof-of-concept experiment is then carried out. A first- and a second-order temporal differentiator with a bandwidth of 25 GHz are experimentally demonstrated. PMID- 19997202 TI - Polarization-independent drop filters based on photonic crystal self-collimation ring resonators. AB - We report here a polarization-independent drop filter (PIDF) based on a photonic crystal self-collimation ring resonator (SCRR). Despite of the large birefringence associated with the polarization-dependent dispersion properties, we demonstrate a PIDF based on multiple-beam interference theory and polarization peak matching (PPM) technique. The PIDF performance was also investigated based on finite-difference time-domain (FDTD) technique, with excellent agreement between the theory and the simulation. For the designed drop wavelength of 1550 nm, the polarization-independent free spectral range is about 36.1 nm, which covers the whole optical communication C-band window. The proposed PIDFs are highly desirable for applications in photonic integrated circuits (PICs). PMID- 19997203 TI - Pump absorption and temperature distribution in erbium-doped double-clad fluoride glass fibers. AB - We investigate diode pump absorption and temperature distribution in three erbium doped double-clad fluoride fibers. Absorption is measured via fluorescence intensity and temperature distribution is measured with thermal imaging. Ray tracing calculations of absorption and heat-equation modeling of temperature distribution are also conducted. We found excellent agreement between measurements and calculations for all fibers. Results indicate that erbium-doped fluoride fiber lasers have already reached maximum output powers allowed under natural convection cooling, with fiber end being the most critical. We propose cooling and fiber design optimizations that may allow an order-of-magnitude further power-scaling. PMID- 19997204 TI - Cyclic Sommerfeld resonances in nanorods at grazing incidences. AB - We investigate electromagnetic scattering from nanoscale wires and reveal the emergence of a family of exotic resonances for source waves close to grazing incidence. These grazing resonances have a much higher Q-bandwidth product and thus, a much higher Q factor and broader bandwidth than the pure plasmonic resonances found in metal nanowires. Furthermore, these grazing resonances are much less susceptible to material losses than surface plasmon resonances. Contrary to the process of exciting surface plasmon resonances, these grazing resonances can arise in both dielectric and metallic nanowires and appear near to the cutoff wavelength of the circular waveguide. This peculiar resonance effect originates from the excitation of long range guided surface waves through the interplay of coherently scattered continuum modes coupled with first-order azimuthal propagating modes of the cylindrical nanowire. These first-order cyclic Sommerfeld waves and associated cyclic Sommerfeld resonances revealed here opens up the possibility of an alternative scheme of enhanced fields with a better merit (higher Q-bandwidth product and lower loss) than conventional surface plasmon resonances in the nano-regime. This nanowire resonance phenomenon can be utilized in broad scientific areas, including: metamaterial designs, nanophotonic integration, nanoantennas, and nanosensors. PMID- 19997205 TI - Iterative bandgap engineering at selected areas of quantum semiconductor wafers. AB - We report on the application of a laser rapid thermal annealing technique for iterative bandgap engineering at selected areas of quantum semiconductor wafers. The approach takes advantage of the quantum well intermixing (QWI) effect for achieving targeted values of the bandgap in a series of small annealing steps. Each QWI step is monitored by collecting a photoluminescence map and, consequently, choosing the annealing strategy of the next step. An array of eight sites, 280 mum in diameter, each emitting at 1480 nm, has been fabricated with a spectral accuracy of better than 2 nm in a standard InGaAs/InGaAsP QW heterostructure that originally emitted at 1550 nm. PMID- 19997206 TI - Superlensing properties of one-dimensional dielectric photonic crystals. AB - We present the experimental observation of the superlensing effect in a slab of a one-dimensional photonic crystal made of tilted dielectric elements. We show that this flat lens can achieve subwavelength resolution in different frequency bands. We also demonstrate that the introduction of a proper corrugation on the lens surface can dramatically improve both the transmission and the resolution of the imaged signal. PMID- 19997207 TI - Cross-correlation between spiral modesand its influence on the overall spatial characteristics of partially coherent beams. AB - The overall spatial structure of a general partially coherent field is shown to be connected with the cross-correlation between the so-called spiral modes, understood as the terms of the spiral-harmonics series expansion of the field. The formalism based on the beam irradiance-moments is used, and the light field is globally described by the beam width, the far-field divergence, the beam quality factor, the orientation of the beam profile and the angular orbital momentum, given as the sum of its asymmetrical and vortex parts. This overall spatial description is expressed in terms of the intermodal coherence features (cross-correlation between spiral modes). The above analytical results are also illustrated by means of an example. PMID- 19997208 TI - Singular topology of optical absorption in biaxial crystals. AB - We show for the first time that biaxial crystals exhibit continua of directions of propagation where the absorption coefficient is the same for the two associated polarization modes. This statement is supported by both calculations and experimental data obtained in Nd:YCOB. PMID- 19997209 TI - An efficient optical knob from slow light to fast light in a coupled nanomechanical resonator-quantum dot system. AB - We theoretically present a highly efficient optical method to obtain slow and fast light in a coupled system consisting of a nanomechanical resonator and quantum dots in terms of mechanically induced coherent population oscillation (MICPO). Turning on or turning off the specific detuning of pump field from exciton resonance, this coupling system can provide us a direct optical way to obtain the slow or fast group velocity without absorption. Our coupling scheme proposed here works as a fast and slow-light knob and may have potential applications in various domains such as optical communication and biology sensor. PMID- 19997210 TI - Enhanced and broadened SRS spectra of toluene mixed with chloroform in liquid core fiber. AB - Stimulated Raman scattering of toluene and its mixture with chloroform is studied in liquid-core optical fiber. The results show a remarkably broadened Raman line of the mixture from about 630 nm to 650 nm by a pumping wavelength at 532 nm, which is assigned to the interaction of CH vibrations of the two liquids. The results suggest that interactions between adjacent vibrations can produce strong and wide Raman spectra in liquid-core fiber which may prove a new simple way for supercontinuum generation. PMID- 19997211 TI - High-efficiency, room-temperature nanosecond Yb:YAG laser. AB - Yb(3+)-doped gain media offer favorable properties for diode-pumped laser amplifiers for high-energy ns-pulses. To reach high optical-to-optical conversion efficiencies at room temperature however, very high and often impractical fluences are required both for pumping and extraction. Low temperature operation offers a solution, but the required cryogenic cooling systems add considerable complexity, bulkiness and cost. Multi-passing both pump and extraction beams through the gain medium is an alternative approach to overcome efficiency limitations at room temperature. In this article we present numerical and experimental results to this effect.We demonstrated ns-pulse output from a diode pumped Yb:YAG amplifier at an energy of 566 mJ and an optical-to-optical efficiency of 20%, which is almost a doubling of the efficiency achieved with ns lasers employing Yb(3+)-doped gain media at this energy level. PMID- 19997212 TI - Photoacoustic imaging of coronary artery stents. AB - Coronary stents are the most commonly used treatment in the United States to repair narrowed or weakened arteries. The ability to visualize the stent during the stenting procedure and post-surgery is crucial to correctly place the stent with respect to the vessel stenosis, and to identify its position within the vessel wall. Current imaging modalities suffer from low contrast, resolution and/or unfavorable artifacts that can inhibit correct visualization of the stent in the artery. We demonstrated the effectiveness of a combined intravascular photoacoustic and intravascular ultrasound imaging method for high resolution and sufficient contrast imaging of commercial stents with respect to the vessel wall. PMID- 19997213 TI - PDM-QPSK: on the system benefits arising from temporally interleaving polarization tributaries at 100Gb/s. AB - We experimentally study, over a dispersion-managed link relying on low chromatic dispersion fibre, the origins of the system benefits provided by temporally interleaving the polarization tributaries of 100Gb/s coherent RZ-PDM-QPSK by half a symbol period. Hence, we demonstrate that the amount of benefits provided by this technique is dependent on the configuration of the WDM transmission system. PMID- 19997214 TI - A high numerical aperture, polymer-based, planar microlens array. AB - We present a novel microfabrication approach for obtaining arrays of planar, polymer-based microlenses of high numerical aperture. The proposed microlenses arrays consist of deformable, elastomeric membranes that are supported by polymer filled microchambers. Each membrane/microchamber assembly is converted into a solid microlens when the supporting UV-curable polymer is pressurized and cured. By modifying the microlens diameter (40-60 microm) and curing pressure (7.5-30 psi), we demonstrated that it is possible to fabricate microlenses with a wide range of effective focal lengths (100-400 microm) and numerical apertures (0.05 0.3). We obtained a maximum numerical aperture of 0.3 and transverse resolution of 2.8 microm for 60 microm diameter microlenses cured at 30 psi. These values were found to be in agreement with values obtained from opto-mechanical simulations. We envision the use of these high numerical microlenses arrays in optical applications where light collection efficiency is important. PMID- 19997215 TI - A bi-stable 2x2 optical switch monolithically integrated with variable optical attenuators. AB - This work presents the development of a novel micromachined 2x2 optical switch monolithically integrated with variable optical attenuators. The proposed device can be easily realized by a standard manufacturing process with single photo mask. The key to realizing this device by such a simple approach is the employment the split-cross-bar (SCB) configuration. With this configuration, the fabrication challenges and layout constraints for accommodating all the sub components of this dual-function device can be completely eliminated. The monolithically-integrated system has four movable mirrors, two bi-stable mechanisms and six actuators. The switching of optical signals is achieved by moving the mirrors attached on the bi-stable mechanisms using four of the actuators. The attenuation of optical power is carried out by moving the mirrors using the other two actuators and the bi-stable mechanisms. Also, only simple in plane motions are needed for these sub-components to achieve all the functionalities. In addition, the adaption of bi-stable mechanisms can reduce the power consumption and simplify the actuation scheme. The measured insertion losses for both channels are about 1.0~1.1 dB, and the cross-talk is less than 60 dB. The attenuation range is about 30 dB for a maximum applied voltage of 20 V. Also, the measured switching time is less than 4 ms. PMID- 19997216 TI - Terahertz ambipolar dual-wavelength quantum cascade laser. AB - Terahertz frequency quantum cascade lasers (THz QCLs) are compact solid-state sources of terahertz radiation that were first demonstrated in 2002. They have a broad range of potential applications ranging from gas sensing and non destructive testing, through to security and medical imaging, with many polycrystalline compounds having distinct fingerprint spectra in the terahertz frequency range. In this article, we demonstrate an electrically-switchable dual wavelength THz QCL which will enable spectroscopic information to be obtained within a THz QCL-based imaging system. The device uses the same active region for both emission wavelengths: in forward bias, the laser emits at 2.3 THz; in reverse bias, it emits at 4 THz. The corresponding threshold current densities are 490 A/cm(2) and 330 A/cm(2), respectively, with maximum operating temperatures of 98K and 120 K. PMID- 19997217 TI - Optically pumped rolled-up InGaAs/GaAs quantum dot microtube lasers. AB - The authors report on the achievement of lasing in rolled-up semiconductor microtubes at room temperature, wherein self-organized InGaAs/GaAs quantum dots are incorporated as the gain medium. The free-standing quantum dot microtubes, with a diameter of approximately 5-6 microm and wall thickness of approximately 100 nm, are formed when the coherently strained InGaAs/GaAs quantum dot heterostructure is selectively released from the GaAs substrate. The devices are characterized by an ultralow threshold (approximately 4 microW) and a minimum intrinsic linewidth of approximately 0.2 - 0.3 nm at room temperature. The multiple lasing modes are analyzed using both the finite-difference time domain method and also a planar dielectric waveguide model. PMID- 19997218 TI - Single-electrode-tuned mode-hop-free tunable laser based on ring coupled cavity. AB - A novel tunable laser which can achieve mode-hop-free single-electrode tuning based on a ring coupled cavity is proposed. The device comprises of a passive ring resonator coupled to two optical waveguides, two far ends of which are terminated by partially reflecting mirrors to constitute a Fabry-Perot (FP) cavity. The waveguide in the ring resonator is divided into a tuning segment and a bias segment. The FP cavity contains an active gain waveguide and passive waveguide segments. A tuning electrode covers the tuning segment inside the ring resonator and the passive waveguide segments in the FP cavity. Mode-hop-free operation with a single-electrode tuning is achieved by properly designing the tuning electrode lengths within the FP cavity and within the ring resonator. Numerical analysis shows that a large mode-hop-free tuning range (>6 nm) with a very high single-mode selectivity can be achieved with an injection current less than 100 mA. PMID- 19997219 TI - Compact-sized and broadband carpet cloak and free-space cloak. AB - Recently, invisible cloaks have attracted much attention due to their exciting property of invisibility, which are based on a solid theory of transformation optics and quasi-conformal mapping. Two kinds of cloaks have been proposed: free space cloaks, which can render objects in free space invisible to incident radiation, and carpet cloaks (or ground-plane cloaks), which can hide objects under the conducting ground. The first free-space and carpet cloaks were realized in the microwave frequencies using metamaterials. The free-space cloak was composed of resonant metamaterials, and hence had restriction of narrow bandwidth and high loss; the carpet cloak was made of non-resonant metamaterials, which have broad bandwidth and low loss. However, the carpet cloak has a severe restriction of large size compared to the cloaked object. The above restrictions become the bottlenecks to the real applications of free-space and carpet cloaks. Here we report the first experimental demonstration of broadband and low-loss directive free-space cloak and compact-sized carpet cloak based on a recent theoretical study. Both cloaks are realized using non-resonant metamaterials in the microwave frequency, and good invisibility properties have been observed in experiments. This approach represents a major step towards the real applications of invisibility cloaks. PMID- 19997220 TI - Laser direct writing using submicron-diameter fibers. AB - In this paper, a novel direct writing technique using submicron-diameter fibers is presented. The submicron-diameter fiber probe serves as a tightly confined point source and it adopts micro touch mode in the process of writing. The energy distribution of direct writing model is analyzed by Three-Dimension Finite Difference Time-Domain method. Experiments demonstrate that submicron-diameter fiber direct writing has some advantages: simple process, 350-nm-resolution (lower than 442-nm-wavelength), large writing area, and controllable width of lines. In addition, by altering writing direction of lines, complex submicron patterns can be fabricated. PMID- 19997221 TI - Near-field optical microscopy with a nanodiamond-based single-photon tip. AB - We introduce a point-like scanning single-photon source that operates at room temperature and offers an exceptional photostability (no blinking, no bleaching). This is obtained by grafting in a controlled way a diamond nanocrystal (size around 20 nm) with single nitrogen-vacancy color-center occupancy at the apex of an optical probe. As an application, we image metallic nanostructures in the near field, thereby achieving a near-field scanning single-photon microscopy working at room temperature on the long term. Our work may be of importance to various emerging fields of nanoscience where an accurate positioning of a quantum emitter is required such as for example quantum plasmonics. PMID- 19997222 TI - Grayscale photomask fabricated by laser direct writing in metallic nano-films. AB - The grayscale photomask plays a key role in grayscale lithography for creating 3D microstructures like micro-optical elements and MEMS structures, but how to fabricate grayscale masks in a cost-effective way is still a big challenge. Here we present novel low cost grayscale masks created in a two-step method by laser direct writing on Sn nano-films, which demonstrate continuous-tone gray levels depended on writing powers. The mechanism of the gray levels is due to the coexistence of the metal and the oxides formed in a laser-induced thermal process. The photomasks reveal good technical properties in fabricating 3D microstructures for practical applications. PMID- 19997223 TI - Optically controllable side-polished fiber attenuator with photoresponsive liquid crystal overlay. AB - This study presents an optically controllable fiber-optic attenuator consisting of side-polished fiber (SPF) with a photoresponsive liquid crystal (LC) overlay operating in the telecommunication wavelength. Attenuation is controlled by a photochemical-induced phase transition of photoresponsive LC, which modulates the evanescent field leaked from the polished area. Before optical field illumination, the photoresponsive LCs are in the light-scattering state and attenuation is high. During photoirradiation, the formation of cis-azobenzene LC disrupts the nematic host and generates a light-transparent state in which the optical loss of the SPF attenuator decreases. The photoinduced tuning range is 15 dB at an environmental temperature of 45 degrees C, and a repeatable and reversible tuning is observed with a response time of less than 5 s. The proposed all-optical controllable attenuator has potential use as an optical signal modulator in an all-fiber telecommunication system. PMID- 19997224 TI - The forces from coupled surface plasmon polaritons in planar waveguides. AB - We analytically investigate the forces due to Surface Plasmon Polariton (SPP) modes between finite and infinitely thick metal slabs separated by an air gap. Using the Drude model and experimentally determined values of the dielectric functions of gold and silver, we study how frequency dispersion and loss in the metals affects the behavior of the SPP modes and the forces generated by them. We calculate the force using the Maxwell Stress Tensor for both the attractive and repulsive modes. PMID- 19997225 TI - In-plane all-photonic transduction with differential splitter using double-step rib waveguide for photonic microcantilever arrays. AB - We report a differential splitter consisting of an asymmetric double-step multimode rib waveguide and a Y-branch splitter for in-plane photonic transduction of photonic microcantilever deflection. Arrays of photonic microcantilevers are integrated with differential splitters and an optical waveguide network to demonstrate uniformity and sensitivity of transduction. Measurement results from multiple arrays indicate a sensitivity of 0.32x10(-3) nm(-1) and minimum detectable deflection of 141 pm for a 3.5 Hz measurement bandwidth. PMID- 19997226 TI - Stable operation of 4 mW nanoseconds radiation at 177.3 nm by second harmonic generation in KBe2BO3F2 crystals. AB - We report the generation of nanoseconds radiation at 177.3 nm with a maximum average power of 34.7 mW by second harmonic generation (SHG) in a 2.06-mm thick KBe(2)BO(3)F(2) (KBBF) crystal pumped with a homemade 4.2 W nanoseconds Nd: YAG laser at 355 nm operating at 10 KHz and 49 ns, which corresponds to an energy efficiency of approximately 0.826%. To our knowledge, it is the highest power generated at 177.3 nm. The dependence of phase matching angle of KBBF on temperature is presented for the first time. We also present the details on the measures for stable operation of a 4 mW nanosecond output at 177.3 nm with a lower pumping power in a thinner 1.37-mm KBBF crystal, and the stable output power is improved by about 20 times compared with previous results. PMID- 19997227 TI - Nonlinear optical properties ofBaAlBO3F2 crystal. AB - We investigated the nonlinear optical properties of new BaAlBO(3)F(2)(BABF) crystal. The high quality BABF is nonhygroscopic and possesses a moderate birefringence suitable for UV light generation. On the basis of its refractive index dispersion curves, it is inferred that BABF has great potential applications nonlinear optical material, notably for UV light generation at 355 nm. In order to characterize its nonlinear optical properties, BABF samples were cut an oriented in phase matching conditions The optical conversion efficiency from 1064 nm to 532 nm was investigated for the first time: up to 49.0% were achieved. The external angular acceptance bandwidth of SHG and THG for 1064 nm pump light was measured. PMID- 19997228 TI - Long period grating working in transition mode as promising technological platform for label-free biosensing. AB - We present the development of a platform for label-free biosensing based on overlayered Long Period Gratings (LPGs) working in transition mode. Nano-scale layers of Polystyrene (PS) with different thicknesses were deposited onto the same LPG to test the performances of the device in different working points of its modified sensitivity characteristic. Adsorption dynamic of biotinylated bovine serum albumin (BBSA) onto the PS overlays was on-line monitored as well as a subsequent streptavidin (SA) binding dynamic on the biotinylated sites of the protein ad-layer. Experimental results show that overlayered LPGs are among the most sensitive refractive index transducers to be employed in label-free biochemical detection and that wide margins of further optimization exist. PMID- 19997229 TI - Laser differential confocal ultra-long focal length measurement. AB - A new laser differential confocal focal-length measurement method is proposed for the measurement of an ultra-long focal-length. The approach proposed uses the property of an axial intensity curve that the absolute zero precisely corresponds to the focus of the objective in a differential confocal focusing system (DCFS) to measure the variation in position of DCFS focus with and without a measured ultra-long focal-length lens (UFL), uses the distance between the two focuses to obtain the UFL focal-length, and thereby achieving the precise measurement of ultra-long focal-length. The method has a high focusing precision, a strong anti interference capability and a short measurement light-path. The theoretical analyses and preliminary experimental results indicate that the relative measurement error is about 0.01% when the method is used for the measurement of back-focus-distance (BFD). PMID- 19997230 TI - Quadratic phase matching in nonlinear plasmonic nanoscale waveguides. AB - We analyze phase matching in metal-dielectric nonlinear structures which support highly localized plasmon polariton modes. We reveal that quadratic phase matching between the plasmon modes of different symmetries becomes possible in planar waveguide geometries. We discuss the example of a nonlinear LiNbO(3) waveguide sandwiched between two silver plates, and demonstrate that second-harmonic generation can be achieved for interacting plasmonic modes. PMID- 19997231 TI - Enhancement of second harmonic generation in one-dimensional nonlinear photonic crystal microcavities. AB - A numerical iteration technique starting from the analytical solution of the fundamental wave and the second harmonic wave propagation equations in undepleted pump approximation is presented for analysis of second harmonic generation in one dimensional nonlinear photonic-crystal microcavities under pump light incidence at an arbitrary angle, accounting for pump depletion. Numerical results are in good agreement with experimental observation and theoretical predication by transfer matrix method. Analysis of the conversion efficiency of second harmonic generation in nonlinear photonic-crystal microcavities shows that dramatic enhancement of second harmonic generation occurs in the microcavities which have the optimal numbers of distributed Bragg reflector layers. The enhancement of nonlinear interactions is ascribed to mode resonance and localization of high intensity fundamental field, corresponding to the defect mode within the forbidden band of the photonic-crystal microcavities. PMID- 19997232 TI - Modeling dispersive coupling and losses of localized optical and mechanical modes in optomechanical crystals. AB - Periodically structured materials can sustain both optical and mechanical excitations which are tailored by the geometry. Here we analyze the properties of dispersively coupled planar photonic and phononic crystals: optomechanical crystals. In particular, the properties of co-resonant optical and mechanical cavities in quasi-1D (patterned nanobeam) and quasi-2D (patterned membrane) geometries are studied. It is shown that the mechanical Q and optomechanical coupling in these structures can vary by many orders of magnitude with modest changes in geometry. An intuitive picture is developed based upon a perturbation theory for shifting material boundaries that allows the optomechanical properties to be designed and optimized. Several designs are presented with mechanical frequency approximately 1-10 GHz, optical Q-factor Qo > 107, motional masses meff approximately 100 femtograms, optomechanical coupling length LOM < 5 microm, and clampinig losses that are exponentially suppressed with increasing number of phononic crystal periods (radiation-limited mechanical Q-factor Qm > 107 for total device size less than 30 microm). PMID- 19997233 TI - Design of an efficient terahertz source using triply resonant nonlinear photonic crystal cavities. AB - We propose a scheme for efficient cavity-enhanced nonlinear THz generation via difference-frequency generation (DFG) processes using a triply resonant system based on photonic crystal cavities. We show that high nonlinear overlap can be achieved by coupling a THz cavity to a doubly-resonant, dual-polarization near infrared (e.g. telecom band) photonic-crystal nanobeam cavity, allowing the mixing of three mutually orthogonal fundamental cavity modes through a chi((2)) nonlinearity. We demonstrate through coupled-mode theory that complete depletion of the pump frequency - i.e., quantum-limited conversion - is possible. We show that the output power at the point of optimal total conversion efficiency is adjustable by varying the mode quality (Q) factors. PMID- 19997234 TI - Sub-100 fs single-walled carbon nanotube saturable absorber mode-locked Yb-laser operation near 1 microm. AB - Transmission- and reflection-type single-walled carbon nanotube saturable absorbers (SWCNT-SAs) were designed and fabricated for passive mode-locking of bulk lasers in the 1 microm spectral range. Mode-locked laser operation based on a diffusion-bonded Yb:KYW/KYW crystal was demonstrated, and pulses as short as 83 fs and 140 fs were achieved applying reflection-type and transmission-type SWCNT SA, respectively. The nonlinear parameters of the absorbers were measured to be in close vicinity to those of a semiconductor saturable absorber mirror for the same wavelength range. Mode-locking performance with SWCNT-SAs and the SESAM was compared utilizing the same cavity, with the SESAM resulting in only slightly shorter pulses of 66 fs duration. The nearly identical performance indicates that well-optimized SWCNT-SAs can substitute SESAMs even in the 1 microm region. PMID- 19997235 TI - Third-harmonic generation via broadband cascading in disordered quadratic nonlinear media. AB - We study parametric frequency conversion in quadratic nonlinear media with disordered ferroelectric domains. We demonstrate that disorder allows realizing broadband third-harmonic generation via cascading of two second-order quasi-phase matched nonlinear processes. We analyze both spatial and polarization properties of the emitted radiation and find the results in agreement with our theoretical predictions. PMID- 19997236 TI - Suppression of time delay signatures of chaotic output in a semiconductor laser with double optical feedback. AB - We experimentally and numerically demonstrate the time delay (TD) signature suppression of chaotic output in a double optical feedback semiconductor laser (DOF-SL) system. Two types of TD signature suppression are demonstrated by adjusting the lengths and the feedback power ratios of the two external cavities. One can significantly eliminate all TD signatures of the DOF-SL system and the corresponding power spectrum distribution becomes quite smooth and flat, the other suppresses one of two TD signatures and remains another one. PMID- 19997237 TI - Surface plasmon reflector based on serial stub structure. AB - Plasmonic reflectors based on serial stub structure are studied in this paper. A general theory of periodic stub structure using transmission line model is developed. The transmission characteristics, e.g., periodicity and symmetry of the spectra, are closely related to the ratio of structure period to stub length. Investigation reveals that the transmission valleys of the spectra could be divided into two categories, which is quite different from conventional Bragg reflectors. Finite-Difference Time-Domain (FDTD) method is used in numerical analysis in this paper. PMID- 19997238 TI - Enhanced degree of temporal coherence through temporal and spatial phase coupling within a focused supercontinuum. AB - In the diffraction of a supercontinuum source, a redistribution of amplitude and phase at the focal region is incurred by the coupling between the supercontinuum and the spatial phase caused by the lens diffraction, making it extremely difficult to predict the focal behaviour. We show that the coupling between the temporal phase of a SC source and the spatial phase from the diffraction by a low numerical aperture (NA) lens causes dramatic alterations in the spectra and the temporal coherence near the focal region, and that this effect is maximized in points of singularity. Furthermore, we show that such an enhancement in temporal coherence can be controlled by the pulse evolution through the photonic crystal fiber, in which nonlinear and disperive effects such as the soliton fission process provides the key phase evolution necessary for dramatically changing the coherence time of the focused electromagnetic wave. PMID- 19997239 TI - Temperature dependent gain characteristics in GaN-based vertical-cavity surface emitting lasers. AB - Temperature dependent gain characteristics and linewidth enhancement factor (alpha-factor) of vertical-cavity surface-emitting lasers with InGaN/GaN multiple quantum wells were studied by measuring the photoluminescence spectra below the threshold condition and analyzed by using the Hakki-Paoli method. The optical gain and differential gain showed a more rapid increase as a function of the injected carriers as temperature decreased. The alpha-factor for the lasing mode was estimated as 2.8 at room temperature and decreased to a value as low as 0.6 at 80 K. PMID- 19997240 TI - Near zero bending loss in a double-trenched bend insensitive optical fiber at 1550 nm. AB - We have developed a new single-mode optical fiber (SMF) which exhibits ultra low bend sensitivity over a wide communication band. The measured mean bending loss at 1550 nm was about 0.0095 dB for a loop of 10 mm diameter. PMID- 19997241 TI - High-accuracy wavefront control for retinal imaging with Adaptive-Influence Matrix Adaptive Optics. AB - We present an iterative technique for improving adaptive optics (AO) wavefront correction for retinal imaging, called the Adaptive-Influence-Matrix (AIM) method. This method is based on the fact that the deflection-to-voltage relation of common deformable mirrors used in AO are nonlinear, and the fact that in general the wavefront errors of the eye can be considered to be composed of a static, non-zero wavefront error (such as the defocus and astigmatism), and a time-varying wavefront error. The aberrated wavefront is first corrected with a generic influence matrix, providing a mirror compensation figure for the static wavefront error. Then a new influence matrix that is more accurate for the specific static wavefront error is calibrated based on the mirror compensation figure. Experimental results show that with the AIM method the AO wavefront correction accuracy can be improved significantly in comparison to the generic AO correction. The AIM method is most useful in AO modalities where there are large static contributions to the wavefront aberrations. PMID- 19997243 TI - Modeling surface plasmon-polariton gain in planar metallic structures. AB - Amplification of the single-interface and long-range surface plasmon-polariton modes is studied in planar metallic structures incorporating gain media formed by Rhodamine 6G dye molecules in solution. We employ a theoretical model that accounts for the nonuniformity of the gain medium close to the metal surface due to position-dependent dipole lifetime and pump irradiance. The results of this model are used as a baseline for a comparative study against two simplified models: one neglects the position-dependent dipole lifetime while the other assumes a uniform gain medium. The discrepancies between the models are explained in terms of the mode overlap with the gain distribution near the metal. For the cases under analysis, the simplified models estimate the required pump irradiance with deviation factors that vary from 1.45 at the lossless conditions to 8 for gains near saturation. The relevance of describing properly the amount o gain interacting with the SPP mode and the role played by the dipole quantum efficiency are discussed. PMID- 19997242 TI - Monte Carlo simulation of photon migration in 3D turbid media accelerated by graphics processing units. AB - We report a parallel Monte Carlo algorithm accelerated by graphics processing units (GPU) for modeling time-resolved photon migration in arbitrary 3D turbid media. By taking advantage of the massively parallel threads and low-memory latency, this algorithm allows many photons to be simulated simultaneously in a GPU. To further improve the computational efficiency, we explored two parallel random number generators (RNG), including a floating-point-only RNG based on a chaotic lattice. An efficient scheme for boundary reflection was implemented, along with the functions for time-resolved imaging. For a homogeneous semi infinite medium, good agreement was observed between the simulation output and the analytical solution from the diffusion theory. The code was implemented with CUDA programming language, and benchmarked under various parameters, such as thread number, selection of RNG and memory access pattern. With a low-cost graphics card, this algorithm has demonstrated an acceleration ratio above 300 when using 1792 parallel threads over conventional CPU computation. The acceleration ratio drops to 75 when using atomic operations. These results render the GPU-based Monte Carlo simulation a practical solution for data analysis in a wide range of diffuse optical imaging applications, such as human brain or small animal imaging. PMID- 19997244 TI - Plasmonic data storage medium with metallic nano-aperture array embedded in dielectric material. AB - We propose a plasmonic data storage medium with a high-transmission metal aperture array embedded in a dielectric material. Bowtie apertures, having an outline of 80 nm and a ridge gap of 30 nm, are arranged in a two dimensional array with a bit pitch of 100 nm and a track pitch of 280 nm. Using the finite differential time domain (FDTD) method, we calculate the exposure power needed to record optical data, the contrast for readability of recorded data, and cross talk between the main track and adjacent tracks. Compared to a conventional blu ray disc, the exposure power needed to record optical data in the proposed plasmonic data storage medium is less than a quarter of the conventional threshold power, and the density of the data storage is about 1.8 times larger. PMID- 19997245 TI - The absolute sensitivity of digital colour cameras. AB - A new and improved method to obtain the average spectral pixel responsivity and the quantum efficiency of Digital Single Lens Reflex (DSLR) cameras is outlined. Two semi-professional cameras, the Nikon D300 and the Canon 40D, are evaluated. The cameras red, green and blue pixel responsivities and quantum efficiency are retrieved by illuminating an integrating sphere with a wavelength tunable monochromator. 31 intensity calibrated monochromatic spectral lines from 4000 to 7000 A, with a bandpass of approximately 12 A, were used as a library to solve the main equations of observation for the cameras. Both cameras have peak sensitivity in the blue and minimum sensitivity in the red. The Canon 40D has blue and green channel sensitivity close to the Nikon D300. The Canon red channel has half the sensitivity of the Nikon camera. PMID- 19997246 TI - Measurement and modeling of high-linearity modified uni-traveling carrier photodiode with highly-doped absorber. AB - The third-order intermodulation distortions of InGaAs/InP modified uni-traveling carrier photodiodes with a highly-doped p-type absorber are characterized. The third-order local intercept point is 55 dBm at low frequency (< 3 GHz) and remains as high as 47.5 dBm up to 20 GHz. The frequency characteristics of the OIP3 are well explained by an equivalent circuit model. PMID- 19997247 TI - Visible-NIR tunable Pr3+-doped fiber laser pumped by a GaN laser diode. AB - We demonstrated continuous wave laser operation with a tunable range of over 300 nm in the visible and near-infrared regions (479-497, 515-548, 597-737, 849-960 nm) using a single 9-cm Pr(3+)-doped ZBLAN fiber pumped by a GaN laser. The total tunable range was 6469 cm(-1), which is wider than that of conventional Ti:sapphire lasers. PMID- 19997248 TI - Polarization-maintaining, high-energy, wavelength-tunable, Er-doped ultrashort pulse fiber laser using carbon-nanotube polyimide film. AB - A high-energy, wavelength-tunable, all-polarization-maintaining Er-doped ultrashort fiber laser was demonstrated using a polyimide film dispersed with single-wall carbon nanotubes. A variable output coupler and wavelength filter were used in the cavity configuration, and high-power operation was demonstrated. The maximum average power was 12.6 mW and pulse energy was 585 pJ for stable single-pulse operation with an output coupling ratio as high as 98.3%. Wide wavelength-tunable operation at 1532-1562 nm was also demonstrated by controlling the wavelength filter. The RF amplitude noise characteristics were examined in terms of their dependence on output coupling ratio and oscillation wavelength. PMID- 19997249 TI - Demonstration of balanced coherent detection using polymer optical waveguide integrated distributed traveling-wave photodetectors. AB - We have demonstrated balanced coherent detection using distributed balanced traveling-wave photodetectors integrated with single mode polymer optical waveguides. Balanced distributed traveling-wave photodetectors having 3 dB bandwidth of 20 GHz exhibited 20 dB signal to noise ratio improvement measured at 15 GHz modulation frequency in a balanced coherent detection demonstration. PMID- 19997250 TI - Dispersion-shifted all-solid high index-contrast microstructured optical fiber for nonlinear applications at 1.55 microm. AB - We report the fabrication of an all-solid highly nonlinear microstructured optical fiber. The structured preform was made by glass extrusion using two types of commercial lead silicate glasses that provide high index-contrast. Effectively single-moded guidance was observed in the fiber at 1.55 microm. The effective nonlinearity and the propagation loss at this wavelength were measured to be 120 W(-1)km(-1) and 0.8 dB/m, respectively. Numerical simulations indicate that the fiber is dispersion-shifted with a zero-dispersion-wavelength of 1475 nm and a dispersion slope of 0.16 ps/nm(2)/km respectively at 1.55 microm. These predictions are consistent with the experimentally determined dispersion of + 12.5 ps/nm/km at 1.55 microm. Tunable and efficient four-wave-mixing based wavelength conversion was demonstrated at wavelengths around 1.55 microm using a 1.5m-length of the fiber. PMID- 19997251 TI - Transmission line model and fields analysis of metamaterial absorber in the terahertz band. AB - Metamaterial (MM) absorber is a novel device to provide near-unity absorption to electromagnetic wave, which is especially important in the terahertz (THz) band. However, the principal physics of MM absorber is still far from being understood. In this work, a transmission line (TL) model for MM absorber was proposed, and with this model the S-parameters, energy consumption, and the power loss density of the absorber were calculated. By this TL model, the asymmetric phenomenon of THz absorption in MM absorber is unambiguously demonstrated, and it clarifies that strong absorption of this absorber under studied is mainly related to the LC resonance of the split-ring-resonator structure. The distribution of power loss density in the absorber indicates that the electromagnetic wave is firstly concentrated into some specific locations of the absorber and then be strongly consumed. This feature as electromagnetic wave trapper renders MM absorber a potential energy converter. Based on TL model, some design strategies to widen the absorption band were also proposed for the purposes to extend its application areas. PMID- 19997252 TI - Terahertz wave polarization analyzer using birefringent materials. AB - We present a terahertz wave polarization analysis method to extract the polarization rotation angle with respect to the horizontal direction. A quartz crystal is used as the polarization analyzer with the optical axis of the crystal fixed at 45 degrees orientation. The polarization angle of the terahertz waves generated from two-color laser-induced gas plasma is extracted by measuring the transmitted ordinary and extraordinary beams. This work demonstrates that low absorbance birefringent materials are good candidates for terahertz polarization analysis. PMID- 19997253 TI - Nonlinear inter-core coupling in triple-core photonic crystal fibers. AB - We have developed a coupled-mode theory to describe light propagation in triple core photonic crystal fibers (PCFs) in the nonlinear regime. The propagation equations including nonlinearities have been solved analytically. The nonlinear coupling for symmetric and asymmetric fibers is modeled for different coupling and nonlinear parameters. We investigate nonlinear inter-core coupling experimentally with a single high intensity pump beam. Optical switching of a signal beam in the presence of a control pump beam at a different wavelength is also demonstrated. The self-phase modulation (SPM) parameter gamma in the triple core PCF studied is determined. PMID- 19997254 TI - Few-cycle oscillator pulse train with constant carrier-envelope- phase and 65 as jitter. AB - We report on an octave-spanning Ti:sapphire laser oscillator stabilized to carrier-envelope-offset frequency zero, generating a pulse train with constant field profile for every pulse. Stabilization is realized using an extended self referenced locking scheme enabling to lock the carrier envelope-offset phase with less than 65 attosecond rms timing jitter. The stabilized system features a pulse repetition rate of 100 MHz with pulses as short as 4.5 fs and 220 mW average output power. With this laser system it was possible for the first time to demonstrate a spectral interference pattern of 1011 oscillator pulses in an out of-loop f-to-2f-interferometer. PMID- 19997255 TI - Photonic crystal structures in ion-sliced lithium niobate thin films. AB - We report on the first realization of photonic crystal structures in 600-nm thick ion-sliced, single-crystalline lithium niobate thin films bonded on a lithium niobate substrate using adhesive polymer benzocyclobutene (BCB). Focused ion beam (FIB) milling is used for fast prototyping of photonic crystal structures with regular cylindrical holes. Unwanted redeposition effects leading to conically shaped holes in lithium niobate are minimized due to the soft BCB layer underneath. A high refractive index contrast of 0.65 between the lithium niobate thin film and the BCB underlayer enables strong light confinement in the vertical direction. For TE polarized light a triangular photonic crystal lattice of air holes with a diameter of 240 nm and a separation of 500 nm has a photonic bandgap in the wavelength range from 1390 to 1500 nm. Experimentally measured transmission spectra show a spectral power dip for the GK direction of the reci ocal lattice with an extinction ratio of up to 15 dB. This is in good agreement with numerical simulations based on the three-dimensional plane wave expansion (PWE) and the finite-difference time-domain (FDTD) method. PMID- 19997256 TI - Direct near-field optical imaging of UV bowtie nanoantennas. AB - We report near-field optical imaging of bowtie nanoantennas obtained using a UV near-field scanning optical microscope (NSOM). A strong and highly localized UV intensity profile was observed at the antenna gap due to the localized surface plasmon resonance. The relationship of optical field enhancement and antenna size is discussed based on numerical simulations and NSOM experiments. PMID- 19997257 TI - A close-ring pair terahertz metamaterial resonating at normal incidence. AB - We present a systematic study of a close-ring pair based freestanding metamaterial fabricated by double-layer, self-aligned photolithography. Terahertz time-domain spectroscopy transmission measurements and numerical simulations have revealed negative index of refraction in the frequency range of 0.66-0.90 THz under normal wave incidence. The observed resonance behaviors can be well explained by a theoretical circuit model. The electromagnetic properties and the figure of merit of such close-ring metamaterials are also explored in terms of geometrical parameters of the unit cell with a goal of providing optimized design for three-dimensional metamaterials and potential device applications. PMID- 19997258 TI - Effect of multiple wavelengths combination on laser-induced damage in multilayer mirrors. AB - The damage effect of the combined irradiation of 1omega and 3omega in multilayer films was investigated. The experiments were held in both the Laser Induced Damage Threshold (LIDT) mode and the damage probability mode. Moreover, the effect of the laser pre-conditioning was also discussed. It was found that with two wavelengths illumination simultaneously, the number of the sensitive defects still govern the damage probability of the samples, and the energy absorption of the defects to pulse laser is a basic process in causing damage. Additionally, correlative theory models were built to explain the experimental results. PMID- 19997259 TI - Electrically controllable photonic molecule laser. AB - We have studied the coherent intercavity coupling of the evanescent fields of two microdisk terahertz quantum-cascade lasers. The electrically controllable optical coupling of the single-mode operating lasers has been observed for cavity spacings up to 30 mum. The strongest coupled photonic molecule with 2 mum intercavity spacing allows to conditionally switch the optical emission by the electrical modulation of only one microdisk. The lasing threshold characteristics demonstrate the linear dependence of the gain of a quantum-cascade laser on the applied electric field. PMID- 19997260 TI - Color-tuning and switching optical transport through CdS hybrid plasmonic waveguide. AB - We report the color-tuning and switching optical transport characters of CdS hybrid plasmonic waveguide by using near-field optical microscopy. The guided photoluminescence spectra under various waveguide lengths demonstrate a spectroscopic red-shift for the part of CdS nanoribbon placed on the sapphire substrate and an energy compensation at Ag film. Surface plasmon polariton leakage and radiation are explored by near-field characterizations. Finite difference time domain simulations have good agreement with the experimental observations of subwavelength confinement and propagation. With a strong end facet emission, the suggested hybrid plasmonic waveguide can serve as a color changeable optical nanosource in integrated photonic devices. PMID- 19997261 TI - Unusual transmission bands of one-dimensional photonic crystals containing single negative materials. AB - Unusual transmission bands are found in one-dimensional photonic crystals composed of alternating layers of positive-index materials and single-negative (permittivity- or permeability-negative) materials. By varying the thicknesses of the positive-index material layers, the number and central frequencies of these transmission bands can be tuned. On the other hand, by varying the thicknesses of the single-negative material layers, only the bandwidths of these transmission bands will change. Furthermore, omnidirectional transmission bands for TE or TM polarization can be realized from these periodic photonic structures. PMID- 19997262 TI - Real-time visualization of Karman vortex street in water flow field by using digital holography. AB - The Karman vortex street generated behind a circular cylinder in water flow field is displayed and analyzed in real time by means of digital holography. Using a modified Mach-Zehnder interferometer, a digital hologram of the flow field in still state and then a video of continuous digital holograms in flowing state are recorded at 14.6 frames per second by a CCD camera, respectively. A series of sequential phase maps of the flow field are numerically reconstructed from the holograms in different states above based on double-exposure holographic interferometry. By seriating these phase maps, the shape and evolution of Karman vortex street can be displayed in real time in the form of a movie. For comparison, numerical simulation of the Karman vortex street under the boundary conditions adopted in the experiment is also presented, and the consistent results indicate that the experimental observation of Karman vortex street by using digital holography is successful and feasible. PMID- 19997263 TI - Enhancing the light transmission of plasmonic metamaterials through polygonal aperture arrays. AB - While plasmonic metamaterials find numerous applications in the field of nanophotonic devices, a device may work as a normal or plasmonic device, depending on whether it operates at the resonance mode. In this paper, the extraordinary light transmission through coaxial polygonal aperture arrays, including circle, hexagon, square, and triangle geometries, is studied using FDTD simulation. Circular, hexagonal and squared aperture arrays have similar high transmission rate, while triangular aperture array has considerably lower transmission rate. It is found that the transmission peaks reflect the resonance modes propagating along the direction of neighboring apertures. We hence rearrange the apertures from square lattice to triangle lattice to obtain a uniform resonance mode along the neighboring apertures. This leads to enhanced light transmission. The study gains understanding of new properties of the metamaterials based on plasmonic resonance. PMID- 19997265 TI - Spatial and spectral beam shaping with space-variant guided mode resonance filters. AB - Novel all-dielectric beam shaping elements were developed based on guided mode resonance (GMR) filters. This was achieved by spatially varying the duty cycle of a hexagonal-cell GMR filter, to locally detune from the resonant condition, which resulted in modified wavelength dependent reflection and transmission profiles, across the device aperture. This paper presents the design, fabrication, and characterization of the device and compares simulations to experimental results. PMID- 19997264 TI - Electrically-pumped compact hybrid silicon microring lasers for optical interconnects. AB - We demonstrate an electrically-pumped hybrid silicon microring laser fabricated by a self-aligned process. The compact structure (D = 50 microm) and small electrical and optical losses result in lasing threshold as low as 5.4 mA and up to 65 degrees C operation temperature in continuous-wave (cw) mode. The spectrum is single mode with large extinction ratio and small linewidth observed. Application as on-chip optical interconnects is discussed from a system perspective. PMID- 19997266 TI - Probing the dynamic differential stiffness of dsDNA interacting with RecA in the enthalpic regime. AB - RecA plays a central role in homologous recombination of DNA. When RecA combines with dsDNA to form RecA-dsDNA nucleofilament, it unwinds dsDNA and changes its structure. The unwinding length extension of a DNA segment interacting with RecA has been studied by various techniques, but the dynamic differential stiffness of dsDNA conjugating with RecA has not been well characterized. We applied oscillatory optical tweezers to measure the differential stiffness of dsDNA molecules, interacting with RecA, as a function of time at a constant stretching force of 33.6pN. The values of the differential stiffness of DNA (for stretching force in the range of 20.0pN to 33.6pN) measured by oscillatory optical tweezers, both before and after its interaction with RecA, are consistent with those measured by stationary optical tweezers. In the dynamic measurement, we have shown that the association (or binding) rate increases with higher concentration of RecA; besides, we have also monitored in real-time the dissociation of RecA from the stretched RecA-dsDNA filament as ATPgammaS was washed off from the sample chamber. Finally, we verified that RecA (I26C), a form of RecA mutant, does not affect the differential stiffness of the stretched DNA sample. It implies that mutant RecA (I26C) does not bind to the DNA, which is consistent with the result obtained by conventional biochemical approach. PMID- 19997267 TI - Nanoimprinted plastic substrates for enhanced surface plasmon resonance imaging detection. AB - Periodic nanostructures fabricated by Nanoimprint Litography (NIL) in low-cost plastic substrates and coated with thin gold film were explored for enhanced surface plasmon resonance imaging (SPRi) detection. Rigorous coupled-wave analysis was used to model the SPRi response of these nanostructured surfaces. Two-dimensional nanogratings and nanogrooves were fabricated on Zeonor 1060R(TM) by NIL and followed by metal deposition. The detection of refractive index changes in the dielectric layer due to bulk medium change, DNA immobilization and DNA hybridization events were monitored using SPRi to assess the corresponding signal amplification. The results indicate target-dependent sensitivity enhancement which is maximized for the detection of biomolecular binding events. The 500 nm period nanogrooves provided a 4 times SPR signal amplification compared to the conventional uniform gold film on SF-11 glass for DNA hybridization detection. Our work demonstrates that the use of nanoimprinted plastic substrates provides a low-cost solution for the SPR-based detection with sensitivity that meets the requirements in practical diagnostic applications. PMID- 19997268 TI - Phase sensitive amplification based on quadratic cascading in a periodically poled lithium niobate waveguide. AB - We propose and demonstrate phase-sensitive amplification based on cascaded second harmonic generation and difference frequency generation within a periodically poled lithium niobate waveguide. Excellent agreement between our numerical simulations and proof-of-principle experiments using a 3-cm waveguide device operating at wavelengths around 1550 nm is obtained. Our experiments confirm the validity and practicality of the approach and illustrate the broad gain bandwidths achievable. Additional simulation results show that the maximum gain/attenuation factor increases quadratically with input pump power, reaching a value of +/- 19.0 dB at input pump powers of 33 dBm for a 3 cm-long waveguide. Increased gains/reduced powers for a fixed gain could be achieved using longer crystals. PMID- 19997269 TI - Optical Scatter Imaging with a digital micromirror device. AB - We had developed Optical Scatter Imaging (OSI) as a method which combines light scattering spectroscopy with microscopic imaging to probe local particle size in situ. Using a variable diameter iris as a Fourier spatial filter, the technique consisted of collecting images that encoded the intensity ratio of wide-to-narrow angle scatter at each pixel in the full field of view. In this paper, we replace the variable diameter Fourier filter with a digital micromirror device (DMD) to extend our assessment of morphology to the characterization of particle shape and orientation. We describe our setup in detail and demonstrate how to eliminate aberrations associated with the placement of the DMD in a conjugate Fourier plane of our microscopic imaging system. Using bacteria and polystyrene spheres, we show how this system can be used to assess particle aspect ratio even when imaged at low resolution. We also show the feasibility of detecting alterations in organelle aspect ratio in situ within living cells. This improved OSI system could be further developed to automate morphological quantification and sorting of non-spherical particles in situ. PMID- 19997270 TI - Spatial phase-shifting characteristic of double grating interferometer. AB - Double grating interferometer is usually used to achieve phase information from distorted wave front by its temporal phase-shifting characteristic. In this paper, the spatial phase-shifting characteristic of double grating interferometer is presented. The explicit intensity distributions of interferograms produced by double gratings are derived with the scalar diffraction theory, and the stable phase shift is found between plus-first, zero and minus-first order interferograms. Results indicate that the phase shift only depends on the grating period and the distance between two gratings if no phase object exists. If phase object exists, it varies on the interferograms. But the phase shifts are equal at any special point of interferograms. In particular, the triple grating interferometer is presented to generate at least four phase shift interferograms simultaneously with the similar method. PMID- 19997271 TI - High reflection mirrors for pulse compression gratings. AB - We report an experimental investigation of high reflection mirrors used to fabricate gratings for pulse compression application at the wavelength of 1.053microm. Two kinds of mirrors are studied: the mixed Metal MultiLayer Dielectric (MMLD) mirrors which combine a gold metal layer with some e-beam evaporated dielectric bilayers on the top and the standard e-beam evaporated MultiLayer Dielectric (MLD) mirrors. Various samples were manufactured, damage tested at a pulse duration of 500fs. Damage sites were subsequently observed by means of Nomarski microscopy and white light interferometer microscopy. The comparison of the results evidences that if MMLD design can offer damage performances rather similar to MLD design, it also exhibits lower stresses; being thus an optimal mirror substrate for a pulse compression grating operating under vacuum. PMID- 19997272 TI - Two-dimensional light confinement in periodic InGaN/GaN nanocolumn arrays and optically pumped blue stimulated emission. AB - Two-dimensional (2D) light diffraction in a uniform array of GaN nanocolumns arranged in a rectangular lattice dramatically enhanced the light intensity at a specific wavelength, indicating the function of 2D distributed feedback (DFB). Here a GaN rectangular-lattice nanocolumn array, which integrated InGaN/GaN multiple quantum wells (MQWs) in the top region of the nanocolumns, was grown by rf-plasma-assisted molecular beam epitaxy (rf-MBE). At a specific wavelength of 471.1 nm, the first observation of stimulated emission from 2D-DFB in an InGaN based nanocolumn array was obtained. The specific wavelength is calculated by the 2D finite-difference time domain (2D-FDTD) method on the assumption of a refractive index dispersion of GaN; a simple expression for specific wavelength, which is a function of the array period L and the hexagon side length S of each nanocolumn, is proposed, which is convenient for producing a simple design of a GaN nanocolumn array structure in a square lattice. PMID- 19997273 TI - Subsurface damage measurement of ground fused silica parts by HF etching techniques. AB - Detection and measurement of subsurface damage of ground optical surfaces are of major concern in the assessment of high damage thresholds fused silica optics for high power laser applications. We herein detail a new principle of SSD measurement based on the utilization of HF acid etching. We also review and compare different subsurface damage (SSD) characterization techniques applied to ground and fine ground fused silica samples. We demonstrate good concordance between the different measurements. PMID- 19997274 TI - A tunable unidirectional surface plasmon polaritons source. AB - A new structure to be used as a tunable unidirectional surface plasmon source is introduced. The structure is composed of two silver films, with a nanoslit fabricated in the top Ag film and lying below is a movable Ag film. The field distribution of the structure is investigated by using the finite-difference time domain(FDTD) method. It is found that the surface plasmon polariton intensity and the splitting ratio change periodically as the bottom film is moved, which is interpreted in terms of surface plasmon polaritons interference in two Fabry Perot(F-P) cavities. The period obtained by the FDTD agrees well with the F-P interferometer model. The surface plasmon polaritons can be unidirectionally excited with a large intensity splitting ratio. PMID- 19997275 TI - Electric currents induced by twisted light in Quantum Rings. AB - We theoretically investigate the generation of electric currents in quantum rings resulting from the optical excitation with twisted light. Our model describes the kinetics of electrons in a two-band model of a semiconductor-based mesoscopic quantum ring coupled to light having orbital angular momentum (twisted light). We find the analytical solution, which exhibits a "circular" photon-drag effect and an induced magnetization, suggesting that this system is the circular analog of that of a bulk semiconductor excited by plane waves. For realistic values of the electric field and material parameters, the computed electric current can be as large as microA; from an applied perspective, this opens new possibilities to the optical control of the magnetization in semiconductors. PMID- 19997276 TI - Spectral behavior of a terahertz quantum-cascade laser. AB - In this paper, the spectral behavior of two terahertz (THz) quantum cascade lasers (QCLs) operating both pulsed and cw is characterized using a heterodyne technique. Both lasers emitting around 2.5 THz are combined onto a whisker contact Schottky diode mixer mounted in a corner cube reflector. The resulting difference frequency beatnote is recorded in both the time and frequency domain. From the frequency domain data, we measure the effective laser linewidth and the tuning rates as a function of both temperature and injection current and show that the current tuning behavior cannot be explained by temperature tuning mechanisms alone. From the time domain data, we characterize the intrapulse frequency tuning behavior, which limits the effective linewidth to approximately 5 MHz. PMID- 19997278 TI - Designs for electromagnetic cloaking a three-dimensional arbitrary shaped star domain. AB - The design of electromagnetic cloaks based on the coordinate transformation requires a suitable geometrical definition of both the internal and the external surfaces of the cloak itself. We describe a straightforward method to design the electromagnetic cloak of a 3d-generic star domain whose surface is defined just by a set of points distributed over it. We also present numerical simulation for the ray tracing of a light beam inside the material calculated for an asymmetric three-dimensional example. PMID- 19997277 TI - Real-time demonstration of 128-QAM-encoded optical OFDM transmission with a 5.25bit/s/Hz spectral efficiency in simple IMDD systems utilizing directly modulated DFB lasers. AB - The feasibility of implementing 128-QAM in off-the-shelf component-based real time optical OFDM (OOFDM) transceivers incorporating advanced channel estimation, on-line performance monitoring and live parameter optimisation, is experimentally investigated, for the first time, in intensity-modulation and direct-detection (IMDD) single-mode fibre (SMF) and multi-mode fibre (MMF) transmission systems involving directly modulated DFB lasers. The highest ever spectral efficiency of 5.25bit/s/Hz is demonstrated successfully in the aforementioned simple systems. Experimental investigations show that, it is feasible to transmit 5.25 Gb/s 128 QAM-encoded OOFDM real-time signals over 25 km MetroCor(TM) SMFs and 500 m 62.5/125 microm OM1 MMFs. The impact of key parameters on the transmission performance of the real-time OOFDM transceivers with 128-QAM encoding are explored, based on which optimum signal clipping ratios are identified. PMID- 19997279 TI - Anomalous near-perfect extraordinary optical absorption on subwavelength thin metal film grating. AB - We demonstrate analytically and numerically that the anomalous extraordinary optical absorption through thin metal grating with air slit arrays can be excited simultaneously by long and short range surface plasmonic modes supported on the surfaces of thin metal film in weak coupling while this absorption can only be excited by short range surface plasmonic mode in strong coupling. In particular, we predict localized plasmonic mode inside air slits holding double-effect to generate the anomalous extraordinary optical absorption when only this mode is present and to suppress this absorption when the pathways excited by localized and surface plasmonic modes are present simultaneously. Furthermore, we present that double-layer metal grating consisting of two identical single-layer metal gratings can be exploited to enhance absorption efficiency more than 99% and 90% for excited localized plasmonic modes and surface plasmonic modes respectively. PMID- 19997280 TI - Coronagraphic mask design using Hermite functions. AB - We introduce a stellar coronagraph that uses a coronagraphic mask described by a Hermite function or a combination of them. It allows the detection of exoplanets providing both deep starlight extinction and high angular resolution. This angular resolution depends on the order of the Hermite function used. An analysis of the coronagraph performance is carried out for different even order masks. Numerical simulations of the ideal case, with no phase errors and perfect telescope pointing, show that on-axis starlight is reduced to very low intensity levels corresponding to a gain of at least 25 magnitudes (10(-10) light intensity reduction). The coronagraphic throughput depends on the Hermite function or combination selected. The proposed mask series presents the same advantages of band limited masks along with the benefit of reducing the light diffracted by the mask border thanks to its particular shape. Nevertheless, for direct detection of Earth-like exoplanets it requires the use of adaptive optics facilities for compensating the perturbations introduced by the atmosphere and by the optical system. PMID- 19997281 TI - Chi2 analysis for estimating the accuracy of optical properties derived from time resolved diffuse-reflectance. AB - Weighted residuals and the reduced chi(2) (chi(R) (2)) value are investigated with regard to their relevance for assessing optical property estimates using the diffusion equation for time-resolved measurements in turbid media. It is shown and explained, for all photon counting experiments including lifetime estimation, why chi(R) (2) increases linearly with the number of photons when there is a model bias. Only when a sufficient number of photons has been acquired, chi(R) (2) is a pertinent value for assessing the accuracy of mu(a) and mu(s)' estimates. It was concluded that chi(R) (2) is of particular interest for cases of small interfiber separation, low-level scattering, strong absorption and incorrect measurement of instrument response function. It was also found that chi(R) (2) is less pertinent for judging mu(a) in case of air boundary effects. PMID- 19997282 TI - Depth-of-focus (DoF) analysis of a 193nm superlens imaging structure. AB - We present a design of a 193 nm superlens imaging structure to enable the printing of 20 nm features. Optical image simulations indicate that the 20 nm resolution is feasible for both the periodic grating feature and the two-slit feature. Nominal depth-of-focus (DoF) position for both features is identified through the image contrast calculations. Simulations show that the two features have a common nominal dose at the nominal DoF to resolve 20 nm critical dimension when a suitable dielectric material is placed between mask and superlens layer. A DoF of micro8 nm is shown to be obtainable for the 20 nm half-pitch grating feature while the respective DoF for the two-slit feature is less than 8 nm which potentially can be enhanced by employing existing lithographic resolution enhancement techniques. PMID- 19997283 TI - Creating an extended focus image of a tilted object in Fourier digital holography. AB - We present a new method to numerically reconstruct images on a tilted plane by digital holography in Fourier configuration. The proposed technique is based on a quadratic deformation of spatial coordinates of the digital hologram. By this approach we demonstrate that it is possible to recover the extended focus image (EFI) of a tilted object in a single reconstruction step from the deformed hologram. PMID- 19997284 TI - Extraordinary optical transmission through multi-layered systems of corrugated metallic thin films. AB - Optical transmission through multi-layered systems of corrugated metallic thin films is investigated by rigorous electromagnetic simulations based on an exact Green tensor method. Compared to a single metal slab of equivalent thickness and volume, it was found that the multi-layered system can significantly impede the field decay, often leading to transmission greater than that expected from the Fabry-Perot resonance-like behavior exhibited by subwavelength slits in a single slab. Extraordinary optical transmission is also observable for systems of layers whose combined thicknesses are much greater than the skin depth of the metal. Structures consisting of up to five layers with a net thickness of 500 nm for the metal films were considered in our study. These findings demonstrate that an appreciable fraction of the optical power that is incident on the thin metal films can be transmitted over distances greater than their skin depth using plasmonic resonances. PMID- 19997285 TI - Coherent transfer of orbital angular momentum to excitons by optical four-wave mixing. AB - We demonstrate the coherent transfer of optical orbital angular momentum (OAM) to the center of mass momentum of excitons in semiconductor GaN using a four-wave mixing (FWM) process. When we apply the optical vortex (OV) as an excitation pulse, the diffracted FWM signal exhibits phase singularities that satisfy the OAM conservation law, which remain clear within the exciton dephasing time (approximately 1ps). We also demonstrate the arbitrary control of the topological charge in the output signal by changing the OAM of the input pulse. The results provide a way of controlling the optical OAM through carriers in solids. Moreover, the time evolution of the FWM with OAM leads to the study of the closed loop carrier coherence in materials. PMID- 19997286 TI - Comparison of wavefront sensor models for simulation of adaptive optics. AB - The new generation of extremely large telescopes will have adaptive optics. Due to the complexity and cost of such systems, it is important to simulate their performance before construction. Most systems planned will have Shack-Hartmann wavefront sensors. Different mathematical models are available for simulation of such wavefront sensors. The choice of wavefront sensor model strongly influences computation time and simulation accuracy. We have studied the influence of three wavefront sensor models on performance calculations for a generic, adaptive optics (AO) system designed for K-band operation of a 42 m telescope. The performance of this AO system has been investigated both for reduced wavelengths and for reduced r(0) in the K band. The telescope AO system was designed for K band operation, that is both the subaperture size and the actuator pitch were matched to a fixed value of r(0) in the K-band. We find that under certain conditions, such as investigating limiting guide star magnitude for large Strehl ratios, a full model based on Fraunhofer propagation to the subimages is significantly more accurate. It does however require long computation times. The shortcomings of simpler models based on either direct use of average wavefront tilt over the subapertures for actuator control, or use of the average tilt to move a precalculated point spread function in the subimages are most pronounced for studies of system limitations to operating parameter variations. In the long run, efficient parallelization techniques may be developed to overcome the problem. PMID- 19997287 TI - Theoretical study of the spurious-free dynamic range of a tunable delay line based on slow light in SOA. AB - We developed a predictive model describing harmonic generation and intermodulation distortions in semiconductor optical amplifiers (SOAs). This model takes into account the variations of the saturation parameters along the propagation axis inside the SOA, and uses a rigorous expression of the gain oscillations harmonics. We derived the spurious-free dynamic range (SFDR) of a slow light delay line based on coherent population oscillation (CPO) effects, in a frequency range covering radar applications (from 40 kHz up to 30 GHz), and for a large range of injected currents. The influence of the high order distortions in the input microwave spectrum is discussed, and in particular, an interpretation of the SFDR improvement of a Mach-Zehnder modulator by CPOs effects in a SOA is given. PMID- 19997288 TI - Dispersion management in femtosecond laser oscillators with highly dispersive mirrors. AB - Recently the manufacture of highly dispersive mirrors with -1300 fs(2) group delay dispersion per reflection was reported. Here we demonstrate the intracavity applicability of these novel mirrors in Ti:sapphire oscillators for the first time, as well as their capability of compensating a substantial amount of material dispersion in the cavity (40 mm fused silica). We also studied the influence of net negative cavity dispersion, realized with these mirrors, on the achievable maximum pulse energy in long-cavity femtosecond oscillators before the onset of anomalous behavior (e.g. multi-pulsing). In addition, we demonstrate a 0.5 GHz Ti:sapphire oscillator the dispersion compensation of which is realized with a single highly dispersive mirror. PMID- 19997289 TI - Spectral phase conjugation via temporal imaging. AB - We experimentally demonstrate wavelength-preserving spectral phase conjugation for compensating chromatic dispersion and self-phase modulation in optical fibers. Our implementation is based on a temporal imaging scheme that uses time lenses realized by broadband four-wave mixing in silicon waveguides. By constructing a temporal analog of a 4-f imaging system, we compensate for pulse distortions arising from second- and third-order dispersion and self-phase modulation in optical fibers. PMID- 19997290 TI - Ho:YLF pumped HBr laser. AB - A Ho:YLF laser pumped HBr molecular laser was developed that produced up to 2.5 mJ of energy in the 4 micron wavelength region. The Ho:YLF laser was fiber pumped using a commercial Tm:fibre laser. The Ho:YLF laser was operated in a single longitudinal mode via injection seeding with a narrow band diode laser which in turn was locked to one of the HBr transitions. The behavior of the HBr laser was described using a rate equation mathematical model and this was solved numerically. Good agreement both qualitatively and quantitatively between the model and experimental results was obtained. PMID- 19997291 TI - Dynamic behavior of an injection-locked quantum-dash Fabry-Perot laser at zero detuning. AB - This work investigates the behavior of a zero-detuned optically-injected quantum dash Fabry-Perot laser as the injected field ratio is increased from near-zero to levels resulting in stable locking. Using a normalized model describing optically injected semiconductor lasers, variations in the slave laser's free-running characteristics are shown to have a strong impact on the coupled system's behavior. The theoretical model is verified experimentally using a high resolution spectrometer. It is found that the quantum-dash laser has the technological advantage of a low linewidth enhancement factor at low bias currents that suppresses undesirable Period-2 and chaotic behavior. Such observations suggest that optically-injected quantum-dash lasers can be used as an enabling component for tunable photonic oscillators. PMID- 19997292 TI - Dual-frequency imaging using an electrically tunable terahertz quantum cascade laser. AB - We report dual-frequency transmission imaging of polycrystalline materials using an electrically tunable terahertz (THz) frequency quantum cascade laser (QCL). Using our system we are able to obtain images at both 3.05 THz and 3.24 THz in a single two-dimensional scan of a sample. By taking the difference of the natural logarithms of the transmission coefficients obtained at each frequency, the difference-attenuation coefficient is determined, and evaluated for samples of lactose monohydrate, glucose monohydrate, sucrose, and the high explosive PETN. We also demonstrate difference-intensity imaging at these frequencies by combining amplitude modulation of the QCL bias with lock-in detection. Owing to the specific molecular absorption spectra of these materials in the THz frequency range, the samples can be distinguished using our technique. PMID- 19997293 TI - Enhanced light emission from erbium doped silicon nitride in plasmonic metal insulator-metal structures. AB - Plasmonic gratings and nano-particle arrays in a metal-insulator-metal structures are fabricated on an erbium doped silicon nitride layer. This material system enables simple fabrication of the structure, since the active nitride layer can be directly grown on metal. Enhancement of collected emission of up to 12 is observed on resonance, while broad off-resonant enhancement is also present. The output polarization behavior of the gratings and nano-particle arrays is investigated and matched to plasmonic resonances, and the behavior of coupled modes as a function of inter-particle distance is also discussed. PMID- 19997294 TI - Tilted fiber grating accelerometer incorporating an abrupt biconical taper for cladding to core recoupling. AB - We demonstrate a compact power-referenced fiber-optic accelerometer using a weakly tilted fiber Bragg grating (TFBG) combined with an abrupt biconical taper. The electric-arc-heating induced taper is located a short distance upstream from the TFBG and functions as a bridge to recouple the TFBG-excited lower-order cladding modes back into the fiber core. This recoupling is extremely sensitive to microbending. We avoid complex wavelength interrogation by simply monitoring power change in reflection, which we show to be proportional to acceleration. In addition, the Bragg resonance is virtually unaffected by fiber bending and can be used as a power reference to cancel out any light source fluctuations. The proposed sensing configuration provides a constant linear response (nonlinearity < 1%) over a vibration frequency range from DC to 250 Hz. The upper vibration frequency limit of measurement is determined by mechanical resonance, and can be tuned by varying the sensor length. The tip-reflection sensing feature enables the sensor head to be made small enough (20~100 mm in length and 2 mm in diameter) for embedded detection. The polymer-tube-package makes the sensor sufficiently stiff for in-field acceleration measurement. PMID- 19997295 TI - Ramsey interferometry with an atom laser. AB - We present results on a free-space atom interferometer operating on the first order magnetically insensitive |F = 1,mF = 0) --> |F = 2,mF = 0) ground state transition of Bose-condensed (87)Rb atoms. A pulsed atom laser is output-coupled from a Bose-Einstein condensate and propagates through a sequence of two internal state beam splitters, realized via coherent Raman transitions between the two interfering states. We observe Ramsey fringes with a visibility close to 100% and determine the current and the potentially achievable interferometric phase sensitivity. This system is well suited to testing recent proposals for generating and detecting squeezed atomic states. PMID- 19997296 TI - High-efficiency intracavity Nd:YVO4?KTA optical parametric oscillator with 3.6 W output power at 1.53 microm. AB - An efficient intracavity KTA optical parametric oscillator (OPO) driven by diode end-pumped acousto-optical Q-switched Nd:YVO(4) laser is demonstrated. The mode mismatch between fundamental cavity and OPO cavity caused by the thermal lens effect in Nd:YVO(4) crystal as the pump power increased was studied. To lessen mode mismatch, a thermal lens-like cavity mirror made of plane BK7 glass induced by idler absorption was introduced into the OPO cavity. Under a diode pump power of 20 W, a maximum 1535nm light output power of 3.6 W was achieved at the pulse repetition rate of 60 kHz, corresponding to a diode-to-signal conversion efficiency of 18%. This is the highest efficiency reported for eye-safe laser based on intracavity OPO. PMID- 19997297 TI - A terahertz plastic wire based evanescent field sensor for high sensitivity liquid detection. AB - A highly sensitive detection method based on the evanescent wave of a terahertz subwavelength plastic wire was demonstrated for liquid sensing. Terahertz power spreading outside the wire core makes the waveguide dispersion sensitive to the cladding index variation, resulting in a considerable deviation of waveguide dispersion. Two liquids with transparent appearances, water and alcohol, are easily distinguished based on the waveguide dispersion, which is consistent with theoretical predictions. A melamine alcohol solution with various concentrations is identified successfully, and the detection limit is up to 20ppm, i.e. equivalent to the index variation on the order of 0.01. PMID- 19997298 TI - A terahertz time-domain spectrometer for simultaneous transmission and reflection measurements at normal incidence. AB - We present a versatile terahertz time-domain spectrometer which allows reflection measurements at normal incidence and double pass transmission measurements in a single experimental setup. Two different modes for transmission measurements are demonstrated for precise measurements of transparent high or low refractive index materials, respectively. The refractive indices and absorption coefficients of cesium iodide, potassium bromide, sodium chloride, polytetrafluoroethylene (PTFE, Teflon), and silicon have been measured in the frequency range between 1.4 and 4.7 THz. The parameters of the Lorentz oscillator functions describing the phonon polariton dispersions of CsI and KBr have been determined. PMID- 19997299 TI - Higher-order-mode assisted silicon-on-insulator 90 degree polarization rotator. AB - We propose and analyze a 90 degrees polarization rotator based on wave coupling through an intermediate, multimode, axially uniform waveguide. The coupling efficiency of the x- and y-polarized fundamental modes between the horizontal and vertical rectangular waveguides is remarkably enhanced with the help of the TE(01) mode in the multimode waveguide. The polarization rotator has a very short (21-microm) conversion length with a 17.22 dB extinction ratio. It also exhibits a 68-nm bandwidth for polarization conversion efficiency above 90%. PMID- 19997300 TI - All-fiber CARS microscopy of live cells. AB - Using an all-fiber laser system consisting of a femtosecond Er/Yb fiber oscillator as the pump and an ultra-highly nonlinear fiber for Stokes generation, we demonstrate multimodal (TPF+SHG+CARS) non-linear optical microscopy of both tissue samples and live cells. Multimodal imaging was successfully performed with pixel dwell times as short as 4 micros at low laser powers (< 40 mW total). PMID- 19997301 TI - Generation of double-scale femto/pico-second optical lumps in mode-locked fiber lasers. AB - We observed generation of stable picoseconds pulse train and double-scale optical lumps with picosecond envelope and femtosecond noise-like oscillations in the same Yb-doped fiber laser with all-positive-dispersion cavity mode-locked due to the effect of non-linear polarization evolution. In the noise-like pulse generation regime the auto-correlation function has a non-usual double (femto- and picosecond) scale shape. We discuss mechanisms of laser switching between two operation regimes and demonstrate a good qualitative agreement between experimental results and numerical modeling based on modified nonlinear Schrodinger equations. PMID- 19997302 TI - The characterization of GH shifts of surface plasmon resonance in a waveguide using the FDTD method. AB - We have explicated the Goos-Hanchen (GH) shift in a mum-order Kretchmann-Raether configuration embedded in an optical waveguide structure by using the finite difference time-domain method. For optical waveguide-type surface plasmon resonance (SPR) devices, the precise derivation of the GH shift has become critical. Artmann's equation, which is accurate enough for bulk optics, is difficult to apply to waveguide-type SPR devices. This is because Artmann's equation, based on the differentiation of the phase shift, is inaccurate at the critical and resonance angles where drastic phase changes occur. In this study, we accurately identified both the positive and the negative GH shifts around the incidence angle of resonance. In a waveguide-type Kretchmann-Raether configuration with an Au thin film of 50 nm, positive and negative lateral shifts of -0.75 and + 1.0 microm are obtained on the SPR with the incident angles of 44.4 degrees and 47.5 degrees, respectively, at a wavelength of 632.8 nm. PMID- 19997303 TI - Average BER analysis of SCM-based free-space optical systems by considering the effect of IM3 with OSSB signals under turbulence channels. AB - In this paper, we derive the average bit error rate (BER) of subcarrier multiplexing (SCM)-based free space optics (FSO) systems using a dual-drive Mach Zehnder modulator (DD-MZM) for optical single-sideband (OSSB) signals under atmospheric turbulence channels. In particular, we consider the third-order intermodulation (IM3), a significant performance degradation factor, in the case of high input signal power systems. The derived average BER, as a function of the input signal power and the scintillation index, is employed to determine the optimum number of SCM users upon the designing FSO systems. For instance, when the user number doubles, the input signal power decreases by almost 2 dBm under the log-normal and exponential turbulence channels at a given average BER. PMID- 19997304 TI - Y-branch integrated dual wavelength laser diode for microwave generation by sideband injection locking. AB - A Y-branch integrated dual wavelength laser diode is fabricated for optical microwave generation based on the principle of sideband injection locking. The device integrates a master laser and a slave laser with associated Y-branch coupler. By directly modulating the master laser near its relaxation resonance frequency, multiple sidebands are generated due to enhanced modulation nonlinearity. Beat signal with high spectral purity is obtained by injection locking the slave laser to one of the modulation sidebands. A millimeter-wave carrier of 42-GHz with a phase noise of -94.6 dBc/Hz at 10 kHz offset is demonstrated. PMID- 19997305 TI - A generalized reference-plane-based calibration method in optical triangular profilometry. AB - In this paper, a generalized reference-plane-based calibration method is proposed in optical triangular profilometry by exploring projection ray tracing method and image ray tracing method. The pin-hole camera model is used to model the camera and the projector, and parallel planes model is used to model the reference and test planes. The camera, projector, and planes can be in arbitrary positions and arbitrary directions. The reciprocal of the height and the reciprocal of the phase shift (or pixel position vertical distance) are in linear relationship. Experiments are conducted to verify the proposed method. PMID- 19997306 TI - Vertical plasmonic Mach-Zehnder interferometer for sensitive optical sensing. AB - Vertical plasmonic Mach-Zehnder Interferometers are investigated theoretically and experimentally, and their potential for ultra-sensitive optical sensing is discussed. Plasmonic interferences arise from coherently coupled pairs of subwavelength slits, illuminated by a broadband optical source, and this interference modulates the intensity of the far-field scattering spectrum. Experimental results, obtained using a simple experimental setup, are presented to validate theoretically predicted interferences introduced by the surface plasmon modes on top and bottom surfaces of a metal film. By observing the wavelength shift of the peaks or valleys of the interference pattern, this highly compact device has the potential to achieve a very high sensitivity relative to other nanoplasmonic architectures reported. PMID- 19997307 TI - Two-dimensional periodic structure induced by single-beam femtosecond laser pulses irradiating titanium. AB - A novel two-dimensional (2-D) periodic structure on the ablated hole wall is formed by using the method of single-beam femtosecond laser pulses irradiating a titanium target. This 2-D structure on the ablated surface, after consecutive irradiation of 10000 pulses, presents different spatial periods which characters a 1500 nm period along the hole wall in the vertical direction and a 400 nm period in the azimuthal direction of the hole. The reported experimental results can be well interpreted by a prophetic theoretical model of Bonch-Bruevich (A. M. Bonch-Bruevich et al., Opt. Eng. 31, 718 (1992)). PMID- 19997308 TI - NanoSi low loss horizontal slot waveguides coupled to high Q ring resonators. AB - Si-based horizontal slot waveguides coupled to ring resonators have been fabricated and characterised. The central layer of the slot has been filled by Silicon nanocrystals (Si-nc) obtained by deposition of silicon rich silicon oxide and then thermal annealing. A comparison of various deposition and annealing parameters to form the Si-nc is performed. Propagation losses as low as 3 dB/cm and ring resonator quality factor of 30,000 have been achieved at 1550 nm. PMID- 19997309 TI - A generalized model of maximizing the sensitivity in intensity-interrogation surface plasmon resonance biosensors. AB - Intensity interrogation of surface plasmon resonance (IISPR) biosensors possesses the greatest sensitivity beyond other interrogations and is operated at a fixed incident angle to enable real-time analysis without time delay, so that it promises excellent performance in biological/chemical detection and SPR imaging systems. Here we provide a general model to describe its sensitivity based on Lorentz equation and unveil the relation between the sensitivity and the metal thickness. This model presents the dependency between sensitivity and metal thickness, and the optimal thickness of gold layers to maximize the sensitivity in our experiment is 53 nm that agrees well in both measurement and simulation. This general model can be further applied in other intensity-interrogation SPR configurations as a design rule for sensing and imaging applications. PMID- 19997311 TI - Silicon dual-ring modulator. AB - A silicon electro-optic modulator based on coupled microring resonators is proposed. The dual-ring modulator has wide optical bandwidth, high extinction ratio and low insertion loss. A simple integrated circuit connecting the two rings allows for fast modulation speed of >40 Gbit/s. This device has both improved optical and electrical performance over a single-ring modulator. The dual-ring modulator can provide a feedback signal for active control of the modulation wavelength. PMID- 19997310 TI - Optical fiber magnetic field sensors with TbDyFe magnetostrictive thin films as sensing materials. AB - Different from usually-used bulk magnetostrictive materials, magnetostrictive TbDyFe thin films were firstly proposed as sensing materials for fiber-optic magnetic field sensing characterization. By magnetron sputtering process, TbDyFe thin films were deposited on etched side circle of a fiber Bragg Grating (FBG) as sensing element. There exists more than 45pm change of FBG wavelength when magnet field increase up to 50 mT. The response to magnetic field is reversible, and could be applicable for magnetic and current sensing. PMID- 19997312 TI - Coherent anti-Stokes Raman scattering of two-phonon complexes in diamond. AB - Coherent anti-Stokes Raman scattering (CARS) associated with the second-order Raman process has been confirmed in diamond. Frequency dependence of CARS was analyzed using two-color synchronously-generated picosecond pulses. Resonance enhancement of chi((3)) was observed at an energy shift corresponding to the overtone Sigma branch (K point) phonon, as well as the fundamental Raman-active Gamma((25+)) phonon. The excitation spectrum of CARS at the two-phonon resonance region showed dispersive dependence, reflecting the interference of resonant CARS and nonresonant four-wave mixing scattering. PMID- 19997314 TI - An explicit formula for metal wire plasmon of terahertz wave. AB - An explicit formula for metal wire plasmon of terahertz wave is analytically derived. The derivation is based on the huge relative permittivities of nonmagnetic metals in the spectral region of terahertz wave, some important properties of modified Bessel functions, and a suitable Taylor expansion. The obtained formula is further checked by many numerical tests. We find that, for all 11 tested nonmagnetic metals, for the whole spectral region of terahertz wave, and for the wide radius range from 10 microm to infinity, the relative deviation for the effective index is always smaller than 5%. This good agreement clearly shows that the derived expression can be conveniently used for the analysis and design of metal wire plasmon of terahertz wave. PMID- 19997313 TI - Comparative studies for Cr4+:YAG crystal and AlGaInAs semiconductor used as a saturable absorber in Q-switched Yb-doped fiber lasers. AB - We demonstrate comparative studies for Cr(4+):YAG crystal and AlGaInAs quantum well (QW) used as a saturable absorbers in passively Q-switched Yb-doped fiber lasers. Both saturable absorbers were designed to be possessed of nearly the same initial transmission. Under a pump power of 24 W, the average output powers were up to 14.4 W and 13.8 W obtained with the AlGaInAs QWs and with the Cr(4+):YAG crystal, respectively. The maximum pulse energies obtained with the Cr(4+):YAG crystal and with the AlGaInAs QWs were found to be 0.35 mJ and 0.45 mJ, respectively. PMID- 19997315 TI - Sub-100 W picosecond output from a phase-conjugate Nd:YVO4 bounce amplifier. AB - We demonstrated >80 W picosecond output at a pulse repetition frequency of 100 MHz from a dual Nd:YVO(4) amplifier laser system consisting of a phase-conjugate Nd:YVO(4) bounce amplifier combined with a second diode-side-pumped Nd:YVO(4) bounce amplifier. The output exhibited high quality spatial form with M(2) < 1.8 and a pulse duration (FWHM) of 9.2 ps. A peak power of >7.4 MW with an average power of 78.5 W was also achieved at a pulse repetition frequency of 1.0 MHz. PMID- 19997316 TI - Manipulative depolarization and reflectance spectra of morphologically controlled nano-pillars and nano-rods. AB - Depolarization of sub-mum-high Si nano-pillar/nano-rod surface reflectance with morphologically controlled anti-reflection spectrum is demonstrated. Extremely small reflectance dip of 1.5% at 400-450 nm for Si nano-pillars is extraordinary when comparing with Si nano-rods, in which the reflectance vs. L/lambda for Si nano-pillars coincides well with the graded-index multilayer based modeling spectrum. Alternatively, Si nano-rods preserve its flattened reflectance spectrum up to 1700 nm, whereas the Si nano-pillar surface reflectance monotonically increases to approach that of bulk Si. The destructive interference is only induced on Si nano-pillar surface with larger aspect-ratio > or =15 and small sidewall slope <7 to suppress surface reflectance at blue-green wavelength region. Anomalous depolarization observed from disordered Si nano-pillar/nano-rod surface reflection indicates that TM-mode incidence interacts with more bound electrons than TE-mode to preserve its effective dielectric permittivity less deviated from the bulk Si. The degraded depolarization ratio observed under TE mode incidence which correlates well with a simplified bounded-electron resonance model is elucidated. PMID- 19997317 TI - Mode-locked picosecond pulse generation from an octave-spanning supercontinuum. AB - We generate mode-locked picosecond pulses near 1110 nm by spectrally slicing and reamplifying an octave-spanning supercontinuum source pumped at 1550 nm. The 1110 nm pulses are near transform-limited, with 1.7 ps duration over their 1.2 nm bandwidth, and exhibit high interpulse coherence. Both the supercontinuum source and the pulse synthesis system are implemented completely in fiber. The versatile source construction suggests that pulse synthesis from sliced supercontinuum may be a useful technique across the 1000 - 2000 nm wavelength range. PMID- 19997318 TI - Experimental evaluation of a full-color compact lensless holographic display. AB - An iterative phase retrieval method for a lensless color holographic display using a single light modulator is experimentally validated. The technique involves iterative calculation of a three-plane synthetic hologram which is displayed on a SLM simultaneously lit with three laser beams providing an RGB illumination. Static and animated two-dimensional flicker-free full color images are reconstructed at a fixed position and captured using a high resolution CMOS sensor. The image finesse, color fidelity, contrast ratio and influence of speckles are evaluated and compared with other techniques of holographic color image encoding. The results indicate the technique superior in a case of full color real-life pictures which are correctly displayed by this ultra-compact and simple projection setup. PMID- 19997319 TI - Silicon based total internal reflection bio and chemical sensing with spectral phase detection. AB - Si-based total internal reflection (TIR) bio/chemical sensor presents an attractive alternative to Surface Plasmon Resonance (SPR) technology due to a relatively simple optical arrangement and technological implementation, as well as a relatively easy bio/chemical immobilization on Si/SiO(2) surface with a number of novel attractive applications. This sensor is based on the control of phase difference between p- and s-polarized components of light reflected from Si/air or Si/water interface in TIR geometry and a high sensitivity of the sensor is granted by a high refractive index of Si (3.56 at 1200 nm). We study properties of TIR sensors in a configuration of spectral phase detection and identify conditions of maximal phase sensitive response. We also experimentally show that the detection limit of Si-based TIR sensor can be lowered down to a level of detection of commercially available SPR devices (10(-6) Refractive Index Units, RIU) under the use of a proper low-noisy method of the phase control. The concept of Si-based TIR opens attractive prospects for the miniaturization of sensor devices, taking advantage of the advanced state of development of Si-based microfabrication technologies, while the proposed spectral phase detection scheme offers much easier packaging and calibration steps. PMID- 19997320 TI - Independent control of beam astigmatism and ellipticity using a SLM for fs-laser waveguide writing. AB - We have used a low repetition rate (1 kHz), femtosecond laser amplifier in combination with a spatial light modulator (SLM) to write optical waveguides with controllable cross-section inside a phosphate glass sample. The SLM is used to induce a controllable amount of astigmatism in the beam wavefront while the beam ellipticity is controlled through the propagation distance from the SLM to the focusing optics of the writing set-up. The beam astigmatism leads to the formation of two separate disk-shaped foci lying in orthogonal planes. Additionally, the ellipticity has the effect of enabling control over the relative peak irradiances of the two foci, making it possible to bring the peak irradiance of one of them below the material transformation threshold. This allows producing a single waveguide with controllable cross-section. Numerical simulations of the irradiance distribution at the focal region under different beam shaping conditions are compared to in situ obtained experimental plasma emission images and structures produced inside the glass, leading to a very satisfactory agreement. Finally, guiding structures with controllable cross section are successfully produced in the phosphate glass using this approach. PMID- 19997321 TI - Dual-operation-mode liquid crystal lens. AB - We demonstrate a dual-operation-mode liquid crystal (LC) lens, which is fabricated with the silica nanoparticle-doped (SND) hybrid-aligned nematic (HAN) LC cell. With AC voltage, the cell behaves as a conventional LC lens. The response time of the SND HAN LC lens is faster than that of the conventional LC lens, which is fabricated using the pristine HAN LC cell. This is because that the doped silica nanoparticles may decrease the dielectric relaxation time constant of the cell. The addition of the silica nanoparticles also increases the viscosity of the LC host, suppresses the backflow motion of the LCs and then decreases the response time of the SND LC lens. With DC voltage, the electrophoretic motion of the doped silica nanoparticles and the agglomerate silica networks on the substrate surface cause the SND HAN LC cell to function as a bistable LC lens. PMID- 19997322 TI - A microfluidic refractometric sensor based on gratings in optical fibre microwires. AB - In this paper we investigate a novel method to manufacture gratings in optical fiber microwires and discuss their application to sensing. Gratings can be manufactured by wrapping an optical fiber microwire on a microstructured rod. This method avoids post-processing the thin optical fiber microwire and it has great flexibility: chirping can be realized by designing the air hole size and position in the microstructured rod. By exploiting the large evanescent field in an inner channel, microfluidic refractometric sensors with sensitivity > 10(3) nm/RIU can be achieved. PMID- 19997323 TI - Gain-switched holmium-doped fibre laser. AB - We demonstrate the first gain-switched, singly doped, single-mode holmium-doped silicate glass fibre laser that operates at 2.106 microm. Using a gain-switched 1.909-microm thulium-doped fibre laser as the pump source, output pulses of energy 3.2 microJ and pulse duration of 150 ns were generated at 80 kHz and slope efficiency of 44%. Pulse stacking within the holmium-doped fibre laser resulted in significantly shorter 70 ns pulses. PMID- 19997324 TI - Coupling of light from microdisk lasers into plasmonic nano-antennas. AB - An optical dipole nano-antenna can be constructed by placing a sub-wavelength dielectric (e.g., air) gap between two metallic regions. For typical applications using light in the infrared region, the gap width is generally in the range between 50 and 100 nm. Owing to the close proximity of the electrodes, these antennas can generate very intense electric fields that can be used to excite nonlinear effects. For example, it is possible to trigger surface Raman scattering on molecules placed in the vicinity of the nano-antenna, allowing the fabrication of biological sensors and imaging systems in the nanometric scale. However, since nano-antennas are passive devices, they need to receive light from external sources that are generally much larger than the antennas. In this article, we numerically study the coupling of light from microdisk lasers into plasmonic nano-antennas. We show that, by using micro-cavities, we can further enhance the electric fields inside the nano-antennas. PMID- 19997325 TI - Enhanced 2D-image upconversion using solid-state lasers. AB - Based on enhanced upconversion, we demonstrate a highly efficient method for converting a full image from one part of the electromagnetic spectrum into a new desired wavelength region. By illuminating a metal transmission mask with a 765 nm Gaussian beam to create an image and subsequently focusing the image inside a nonlinear PPKTP crystal located in the high intra-cavity field of a 1342 nm solid state Nd:YVO(4) laser, an upconverted image at 488 nm is generated. We have experimentally achieved an upconversion efficiency of 40% under CW conditions. The proposed technique can be further adapted for high efficiency mid-infrared image upconversion where direct and fast detection is difficult or impossible to perform with existing detector technologies. PMID- 19997326 TI - Propagation losses in undoped and n-doped polycrystalline silicon wire waveguides. AB - Polycrystalline silicon (polySi) wire waveguides with width ranging from 200 to 500 nm are fabricated by solid-phase crystallization (SPC) of deposited amorphous silicon (a-Si) on SiO(2) at a maximum temperature of 1000 degrees C. The propagation loss at 1550 nm decreases from 13.0 to 9.8 dB/cm with the waveguide width shrinking from 500 to 300 nm while the 200-nm-wide waveguides exhibit quite large loss (>70 dB/cm) mainly due to the relatively rough sidewall of waveguides induced by the polySi dry etch. By modifying the process sequence, i.e., first patterning the a-Si layer into waveguides by dry etch and then SPC, the sidewall roughness is significantly improved but the polySi crystallinity is degraded, leading to 13.9 dB/cm loss in the 200-nm-wide waveguides while larger losses in the wider waveguides. Phosphorus implantation causes an additional loss in the polySi waveguides. The doping-induced optical loss increases relatively slowly with the phosphorus concentration increasing up to 1 x 10(18) cm(-3), whereas the 5 x 10(18) cm(-3) doped waveguides exhibit large loss due to the dominant free carrier absorption. For all undoped polySi waveguides, further 1-2 dB/cm loss reduction is obtained by a standard forming gas (10%H(2) + 90%N(2)) annealing owing to the hydrogen passivation of Si dangling bonds present in polySi waveguides, achieving the lowest loss of 7.9 dB/cm in the 300-nm-wide polySi waveguides. However, for the phosphorus doped polySi waveguides, the propagation loss is slightly increased by the forming gas annealing. PMID- 19997327 TI - Hybridized nanocavities as single-polarized plasmonic antennas. AB - We experimentally demonstrate that hybridized nanocavities in optically thick metal films radiate in coherence, and act as an efficient single-polarized plasmonic nano-antenna array. We employ propagating and localized plasmons to enhance polarization control along one axis, with total suppression of the perpendicular polarization component. The relationship between the near-field and far-field radiation is established through a quasi-static model connecting the individual nano-antenna behavior to the phenomenon of extraordinary light transmission. Hybridized nanocavity antennas, with length scales below the conventional diffraction limit, present opportunities for potential applications in photovoltaics, optoelectronic devices and optical sensors. PMID- 19997328 TI - High-Q double-disk microcavities for cavity optomechanics. AB - We design a double-disk microcavity consisting of a pair of silica microdisks separated by a nanoscale gap region on a silicon chip for cavity optomechanics. We show that this type of photonic structure can provide a per-photon gradient force with a magnitude much larger than for scattering-force-based structures. Moreover, this device provides for nearly independent optimization of optical and mechanical properties. We present the processing details of fabricated devices. PMID- 19997329 TI - Demonstration of an optical frequency synthesizer with zero carrier-envelope offset frequency stabilized by the direct locking method. AB - We developed an optical frequency synthesizer (OFS) with the carrier-envelope offset frequency locked to 0 Hz achieved using the "direct locking method." This method differs from a conventional phaselock method in that the interference signal from a self-referencing f-2f interferometer is directly fed back to the carrier-envelope-phase control of a femtosecond laser in the time domain. A comparison of the optical frequency of the new OFS to that of a conventional OFS stabilized by a phase-lock method showed that the frequency comb of the new OFS was not different to that of the conventional OFS within an uncertainty of 5.68x10(-16). As a practical application of this OFS, we measured the absolute frequency of an acetylene-stabilized diode laser serving as an optical frequency standard in optical communications. PMID- 19997330 TI - High power pulsed fiber MOPA system incorporating electro-optic modulator based adaptive pulse shaping. AB - We demonstrate active pulse shaping using an Electro-Optic Modulator in order to compensate the pulse shaping effects caused by Gain Saturation in a high power Yb doped fiber amplifier chain and to generate various custom-defined output pulse shapes. Square, step and smooth pulse shapes are achieved, with mJ pulse energies. Use of a modulator to shape pulses rather than direct modulation of the diode drive current allows us to eliminate undesired transients due to laser start up dynamics. The required shaping is calculated based on a simple measurement of amplifier performance, and does not require detailed modeling of the amplifier dynamics. PMID- 19997331 TI - Monolithic integration of elliptic-symmetry diffractive optical element on silicon-based 45 degrees micro-reflector. AB - A monolithically integrated micro-optical element consisting of a diffractive optical element (DOE) and a silicon-based 45 degrees micro-reflector is experimentally demonstrated to facilitate the optical alignment of non-coplanar fiber-to-fiber coupling. The slanted 45 degrees reflector with a depth of 216 microm is fabricated on a (100) silicon wafer by anisotropic wet etching. The DOE with a diameter of 174.2 microm and a focal length of 150 microm is formed by means of dry etching. Such a compact device is suitable for the optical micro system to deflect the incident light by 90 degrees and to focus it on the image plane simultaneously. The measured light pattern with a spot size of 15 microm has a good agreement with the simulated result of the elliptic-symmetry DOE with an off-axis design for eliminating the strongly astigmatic aberration. The coupling efficiency is enhanced over 10-folds of the case without a DOE on the 45 degrees micro-reflector. This device would facilitate the optical alignment of non-coplanar light coupling and further miniaturize the volume of microsystem. PMID- 19997332 TI - Single-anchor support and supercritical CO2 drying enable high-precision microfabrication of three-dimensional structures. AB - In high-precision two-photon microfabrication of three-dimensional (3-D) polymeric microstructures, supercritical CO(2) drying was employed to reduce surface tension, which tends to cause the collapse of micro/nano structures. Use of supercritical drying allowed high-aspect ratio microstructures, such as micropillars and cantilevers, to be fabricated. We also propose a single-anchor supporting method to eliminate non-uniform shrinkage of polymeric structures otherwise caused by attachment to the substrate. Use of this method permitted frame models such as lattices to be produced without harmful distortion. The combination of supercritical CO(2) drying and the single-anchor supporting method offers reliable high-precision microfabrication of sophisticated, fragile 3-D micro/nano structures. PMID- 19997333 TI - Enhanced 2.0 microm emission and gain coefficient of transparent glass ceramic containing BaF2: Ho3+,Tm3+ nanocrystals. AB - Transparent glass ceramic containing BaF(2):Ho(3+),Tm(3+) nanocrystals has been prepared by melt quenching and subsequent thermal treatment. The precipitation of BaF(2) nanocrystals was confirmed by X-ray diffraction and high-resolution transmission electron microscopy. Intense 2.0 microm fluorescence originating from Ho(3+): (5)I(7) --> (5)I(8) transition was achieved upon excitation with 808 nm laser diode. A large ratio of forward Tm(3+) --> Ho(3+) energy transfer constant to that of backward process indicated high efficient energy transfer from Tm(3+)((3)F(4)) to Ho(3+)((5)I(7)), benefited from the reduced ionic distances of Tm(3+)-Tm(3+) and Tm(3+)-Ho(3+) pairs and low phonon energy environment with the incorporation of rare-earth ions into the precipitated BaF(2) nanocrystals. The results indicate that glass ceramic is a promising candidate material for 2.0 microm laser. PMID- 19997334 TI - An investigation of harmonic generation in liquid media with a mid-infrared laser. AB - We present a harmonic generation experiment using liquid H(2)O and D(2)O interrogated by a mid-infrared, 3.66 mum, laser at a maximum intensity of 8x10(13) W/cm(2). The unique aspects of the experiment include the long wavelength and short (9 cycle-110 fs) pulse duration of the laser as well as the near-resonant excitation of the H(2)O and D(2)O vibrational modes. We observe up to the 13th harmonic order in H(2)O and intensity scaling is consistent with a direct perturbative process up to the 9th harmonic order. Phase matching and resonant absorption are unable to account for the observed differences in harmonic yields between samples. PMID- 19997335 TI - Realization of optical carpets in the Talbot and Talbot-Lau configurations. AB - Talbot and Talbot-Lau effects are frequently used in lensless imaging applications with light, ultrasound, x-rays, atoms and molecules--generally in situations where refractive optical elements are non-existent or not suitable. We here show an experimental visualization of the intriguing wave patterns that are associated with near-field interferometry behind a single periodic diffraction grating under plane wave illumination and which are often referred to as Talbot carpets or quantum carpets. We also show the patterns behind two separated diffraction gratings under nearly-monochromatic but spatially incoherent illumination that illustrate the nature of Talbot-Lau carpets. PMID- 19997336 TI - How much can guided modes enhance absorption in thin solar cells? AB - Absorption enhancement in thin metal-backed solar cells caused by dipole scatterers embedded in the absorbing layer is studied using a semi-analytical approach. The method accounts for changes in the radiation rate produced by layers above and below the dipole, and treats incoherently the subsequent scattering of light in guided modes from other dipoles. We find large absorption enhancements for strongly coupled dipoles, exceeding the ergodic limit in some configurations involving lossless dipoles. An antireflection-coated 100-nm layer of a- Si:H on Ag absorbs up to 87% of incident above-gap light. Thin layers of both strong and weak absorbers show similar strongly enhanced absorption. PMID- 19997337 TI - Light-extraction enhancement of red AlGaInP light-emitting diodes with antireflective subwavelength structures. AB - We demonstrate the enhancement of light extraction in 633 nm AlGaInP light emitting diodes (LEDs) with antireflective subwavelength structures (SWS). From the contour plots by the rigorous coupled wave analysis method, it is found that the reduction of the internal reflection strongly depends on the period of SWS. The Ag nanoparticles formed by thermal dewetting were used as an etch mask for dry etch process to fabricate antireflective SWS on the LED surface. The tapered pillars on the GaP were fabricated, on average, with distances below 200 nm, satisfying the required antireflection condition at the emission wavelength. The improvement in light output power by approximately 26.4% was achieved for the fabricated AlGaInP LEDs with SWS compared to the conventional LEDs due to a strongly reduced Fresnel internal reflection at the GaP/air interface. The improved directionality in the far-field pattern was also obtained due to the directional light extraction enhancement. PMID- 19997338 TI - Ultra-high quality-factor resonators with perfect azimuthal modal-symmetry. AB - We study circular grating resonators (CGRs) which are formed by a central defect surrounded by concentric rings composing a grating and which display perfect azimuthal modal-symmetry. Because of their radial symmetry they exhibit a complete band gap for a minimal index contrast. However, as is the case for all 2D resonators their quality factors are limited by vertical losses. To reduce the vertical losses we introduce a chirp of the grating period by reducing it towards the central defect. The chirped CGRs exhibit drastically improved quality factors of up to tens of millions with a modal volume of a few cubic wavelengths. PMID- 19997339 TI - Rotating single atoms in a ring lattice generated by a spatial light modulator. AB - We demonstrated trapping single neutral Rb atoms in micro traps of an optical ring lattice formed by superposing the +/-l components of the Laguerre-Gaussian mode, and generated by reflecting a single laser beam from a computer controlled spatial light modulator. A single atom in one trap or two atoms with one each in two traps were identified by observing the fluorescence. The trap array loaded with single atoms was rotated by dynamically displaying the hologram animation movie on the modulator. The modulation period in the fluorescence indicates the rotation of one or two single atoms in the lattice. PMID- 19997340 TI - Single-shot holography for depth resolved three dimensional imaging. AB - We introduce a method for depth-resolved photorefractive holographic imaging with potentially extremely short acquisition time for a complete three dimensional (3D) image. By combining the advantages of full-field frequency-domain optical coherence tomography with those of photorefractive holography our concept is capable of obtaining 3D information with only one single shot. We describe the operation principle of our concept and give a first experimental proof of principle. PMID- 19997341 TI - Broadband gradient index microwave quasi-optical elements based on non-resonant metamaterials. AB - Utilizing non-resonant metamaterial elements, we demonstrate that complex gradient index optics can be constructed exhibiting low material losses and large frequency bandwidth. Although the range of structures is limited to those having only electric response, with an electric permittivity always equal to or greater than unity, there are still numerous metamaterial design possibilities enabled by leveraging the non-resonant elements. For example, a gradient, impedance matching layer can be added that drastically reduces the return loss of the optical elements due to reflection. In microwave experiments, we demonstrate the broadband design concepts with a gradient index lens and a beam-steering element, both of which are confirmed to operate over the entire X-band (roughly 8-12 GHz) frequency spectrum. PMID- 19997342 TI - High resolution interferometer with multiple-pass optical configuration. AB - An interferometer having fourteen times higher resolution than a conventional single-pass interferometer has been developed by making multiple-pass optical path. To embody the multiple-pass optical configuration, a two-dimensional corner cube array block was designed, and its symmetric structure minimized the measurement error. The effect from the alignment error and the imperfection of corner cube is calculated as picometer level. An experiment proves that the suggested interferometer has about 45 nm of optical resolution and its nonlinearity is about 0.5 nm in peak-to-valley. PMID- 19997343 TI - The longitudinal offset technique for apodization of coupled resonator optical waveguide devices: concept and fabrication tolerance analysis. AB - In this paper, a novel technique to set the coupling constant between cells of a coupled resonator optical waveguide (CROW) device, in order to tailor the filter response, is presented. The technique is demonstrated by simulation assuming a racetrack ring resonator geometry. It consists on changing the effective length of the coupling section by applying a longitudinal offset between the resonators. On the contrary, the conventional techniques are based in the transversal change of the distance between the ring resonators, in steps that are commonly below the current fabrication resolution step (nm scale), leading to strong restrictions in the designs. The proposed longitudinal offset technique allows a more precise control of the coupling and presents an increased robustness against the fabrication limitations, since the needed resolution step is two orders of magnitude higher. Both techniques are compared in terms of the transmission esponse of CROW devices, under finite fabrication resolution steps. PMID- 19997344 TI - Elongation of femtosecond filament by molecular alignment in air. AB - We study the influence of the molecular alignment on the plasma channel length of femtosecond filament at 800 nm in air. The filament length is observed to be nearly doubled when the high-energy femtosecond laser pulse is properly tuned to match the perpendicular revivals of the molecular alignment, which is different from the low-energy femtosecond laser pulse where the filament is promoted for parallel molecular alignment revivals. These are understood to be the loosened or tightened focusing condition of the propagation of the femtosecond laser pulse by the cross-(de)focusing effect from the prealigned molecules. PMID- 19997345 TI - Angled sidewalls in silicon slot waveguides: conformal filling and mode properties. AB - Effect of angled sidewalls on the filling and properties of silicon slot waveguides is discussed. We demonstrate complete filling of slot waveguide structures with oxide material systems using the atomic layer deposition technique and discuss use of various slot filling materials. Properties of the optical modes in angled-sidewall slot waveguides are studied. Enhanced vertical confinement is obtained with certain waveguide parameters. The reduced effective mode area enhances e.g. nonlinear effects in the waveguide. We discuss the use of atomic layer deposition in realization of filled slot waveguides optimized for all-optical functionalities. PMID- 19997346 TI - Optimization of a polarization imaging system for 3D measurements of transparent objects. AB - This paper presents a multispectral imaging system for 3D reconstruction of transparent objects based on "shape from polarization" technique. The originality of this work relies on a multispectral active lighting system which enables to cope with the two ambiguities on the zenith angle and azimuth angle. A calibration step allows optimising the polarimetric measurements. Example of a reconstructed transparent object is presented. PMID- 19997347 TI - Tests of a practical visible-NIR imaging Fourier transform spectrometer for biological and chemical fluorescence emission measurements. AB - An imaging Fourier transform spectrometer (IFTS) designed for fluorescence emission measurements is reported. The spectral range extension from NIR to visible of the system is realized by using a simple and low-cost optical beam folding position-tracking technique. Spectral resolution as high as 9.78cm( 1)(0.4nm at 632.8nm) and maximum image resolution up to 300x300 pixels are proved by the system tests on its optical performances. Imaging fluorescence spectra acquisition of quantum dot clusters and single 200nm diameter fluorescent beads have demonstrated the system's potential for high throughput imaging spectroscopic measurements of fluorescent biological and chemical samples. PMID- 19997348 TI - Carrier-envelope phase stabilization of a multi-millijoule, regenerative amplifier-based chirped-pulse smplifier system. AB - This article reports on the successful stabilization of the carrier-envelope phase of a 1-kHz laser system that includes a large grating stretcher, a regenerative amplifier, a multipass amplifier and a grating compressor. Phase stability for pulse energies up to 6 mJ is demonstrated using electronic feedback to the oscillator locking electronics as well as feedback via an acousto-optic programmable dispersive filter. PMID- 19997349 TI - Room-temperature photoluminescence in erbium-doped deuterated amorphous carbon prepared by low-temperature MO-PECVD. AB - We report on a novel optical thin film material, erbium-doped deuterated amorphous carbon, fabricated directly on silicon substrate at room-temperature via controlled thermal evaporation of a Metal-Organic compound in a Plasma Enhanced Chemical Vapour Deposition (MO-PECVD) system. High erbium concentrations (up to 2.3 at.%) and room-temperature photoluminescence at 1.54 microm are successfully demonstrated. Concentration quenching due to erbium clustering is reduced by adopting an appropriate MO precursor-Er(tmhd)(3). Another quenching mechanism, caused by non-radiative C-H and O-H vibrational transitions, is shown for the first time to be significantly reduced by deuteration instead of hydrogenation of amorphous carbon. Our results suggest that erbium-doped deuterated amorphous carbon is a promising new class of photonic material for silicon-compatible optoelectronics applications in the technologically important 1.5microm wavelength region. PMID- 19997350 TI - Extremely low power optical bistability in silicon demonstrated using 1D photonic crystal nanocavity. AB - We demonstrate optical bistability in silicon using a high-Q (Q > 105) one dimensional photonic crystal nanocavity at an extremely low 1.6 microW input power that is one tenth the previously reported value. Owing to the device's unique geometrical structure, light and heat efficiently confine in a very small region, enabling strong thermo-optic confinement. We also showed with numerical analyses that this device can operate at a speed of approximately 0.5 micros. PMID- 19997351 TI - Infrared imaging with a wavefront-coded singlet lens. AB - We describe the use of wavefront coding for the mitigation of optical aberrations in a thermal imaging system. Diffraction-limited imaging is demonstrated with a simple singlet which enables an approximate halving in length and mass of the optical system compared to an equivalent two-element lens. PMID- 19997352 TI - Surface microstructuring of Ti plates by femtosecond lasers in liquid ambiences: a new approach to improving biocompatibility. AB - Microstructuring of Ti plates with femtosecond laser pulses is investigated in three different liquids. In these ambiences, complex microstructures with voids and islands are produced on the sample surfaces, whose feature sizes are controlled by the laser parameters. Through adopting supersaturated Hydroxyapatite suspension with higher incident laser fluences, it is for the first time to observe the firm deposition of biocompatible elements Ca-P on the microstructures. At lower laser fluence, only porous structure is present but without additional elements deposition. Both plasma-related ablation under the confinement of liquids and micro-bubbles striking are employed to discuss such structures formation. Tight combining elements Ca-P onto the structured surfaces provide a new way to improve the biocompatibility of body-embedded devices. PMID- 19997353 TI - Electroactive micro and nanowells for optofluidic storage. AB - This paper reports an optofluidic architecture which enables reversible trapping, detection and long term storage of spectrally multiplexed semiconductor quantum dot cocktails in electrokinetically active wells ranging in size from 200nm to 5microm. Here we describe the microfluidic delivery of these cocktails, fabrication method and principal of operation for the wells, and characterize the readout capabilities, storage and erasure speeds, internal spatial signal uniformity and potential storage density of the devices. We report storage and erase speeds of less than 153ms and 30ms respectively and the ability to provide 6-bit storage in a single 200nm well through spectral and intensity multiplexing. Furthermore, we present a novel method for enabling passive long term storage of the quantum dots in the wells by transporting them through an agarose gel matrix. We envision that this technique could find eventual application in fluidic memory or display devices. PMID- 19997354 TI - Direct studies of liquid flows near solid surfaces by total internal reflection fluorescence cross-correlation spectroscopy. AB - We present a new method to study flow of liquids near solid surface: Total internal reflection fluorescence cross-correlation spectroscopy (TIR-FCCS). Fluorescent tracers flowing with the liquid are excited by evanescent light, produced by epi-illumination through the periphery of a high numerical aperture oil-immersion objective. The time-resolved fluorescence intensity signals from two laterally shifted observation volumes, created by two confocal pinholes are independently measured. The cross-correlation of these signals provides information of the tracers' velocities. By changing the evanescent wave penetration depth, flow profiling at distances less than 200 nm from the interface can be performed. Due to the high sensitivity of the method fluorescent species with different size, down to single dye molecules can be used as tracers. We applied this method to study the flow of aqueous electrolyte solutions near a smooth hydrophilic surface and explored the effect of several important parameters, e.g. tracer size, ionic strength, and distance between the observation volumes. PMID- 19997355 TI - Quasi-uniform excitation source for cascade enhancement of SERS via focusing of surface plasmons. AB - A novel surface-enhanced Raman scattering (SERS) excitation source based on focusing of surface plasmons around the center hole of a metal disk for cascaded enhancement is put forward and studied theoretically. The device offers intense SERS excitation with quasi-uniformity and horizontal polarization over a comparatively large hole through the combination of electromagnetic field focusing and hole plasmon resonance. As revealed by finite-difference time-domain (FDTD) method, the intensity spectra and the characteristics of the near field for the wavelength range of 650-1000nm exhibit a number of enhancement modes. Electric field intensity of the optimal mode enhances the SERS signal inside the hole by over four orders. An analytical model was also developed to gain precise interpretation on FDTD results. Our model also reveals the possibility of achieving eight orders of enhancement by optimizing the scale of the disk. In addition to generation of highly optimized hot spots, the large active hole also offers potential applications in fluorescence enhancement and nonlinear spectroscopy. PMID- 19997356 TI - Luminescence from Bi2+-activated alkali earth borophosphates for white LEDs. AB - Bi(2+)-doped MBPO(5) (M = Ba(2+), Sr(2+), Ca(2+)), synthesized in air via solid state reaction, are considered as novel orange and red phosphors for white light emitting diodes with improved colour quality. Absorption of Bi(2+) due to (2)P(1/2)-->(2)S(1/2) and (2)P(1/2)-->(2)P(3/2) could be observed and quantified. Excitation to (2)P(3/2) is accompanied by vibronic sidebands, and corresponding emission behaviour is found. The electron-phonon coupling strength increases in the order M = Ba(2+)-->Sr(2+)-->Ca(2+). In the case of MBPO(5):Bi(2+), one-, two- and even three-phonon sidebands could clearly be observed. The crystal structure of all three compounds belongs to space group P3(1)21. Bi(2+) is incorporated on M(2+) sites, and reduction of Bi(3+) to Bi(2+) occurs for reasons of charge compensation. In accordance with crystallographic data, fluorescence decay behaviour indicates that only one type of Bi(2+)-emission centers is present. PMID- 19997357 TI - Unified perfectly matched layer for finite-difference time-domain modeling of dispersive optical materials. AB - Finite-difference time-domain (FDTD) simulations of any electromagnetic problem require truncation of an often-unbounded physical region by an electromagnetically bounded region by deploying an artificial construct known as the perfectly matched layer (PML). As it is not possible to construct a universal PML that is non-reflective for different materials, PMLs that are tailored to a specific problem are required. For example, depending on the number of dispersive materials being truncated at the boundaries of a simulation region, an FDTD code may contain multiple sets of update equations for PML implementations. However, such an approach is prone to introducing coding errors. It also makes it extremely difficult to maintain and upgrade an existing FDTD code. In this paper, we solve this problem by developing a new, unified PML algorithm that can effectively truncate all types of linearly dispersive materials. The unification of the algorithm is achieved by employing a general form of the medium permittivity that includes three types of dielectric response functions, known as the Debye, Lorentz, and Drude response functions, as particular cases. We demonstrate the versatility and flexibility of the new formulation by implementing a single FDTD code to simulate absorption of electromagnetic pulse inside a medium that is adjacent to dispersive materials described by different dispersion models. The proposed algorithm can also be used for simulations of optical phenomena in metamaterials and materials exhibiting negative refractive indices. PMID- 19997358 TI - Phase and amplitude sensitivities in surface plasmon resonance bio and chemical sensing. AB - We consider amplitude and phase characteristics of light reflected under the Surface Plasmon Resonance (SPR) conditions and study their sensitivities to refractive index changes associated with biological and chemical sensing. Our analysis shows that phase can provide at least two orders of magnitude better detection limit due to the following reasons: (i) Maximal phase changes occur in the very dip of the SPR curve where the vector of probing electric field is maximal, whereas maximal amplitude changes are observed on the resonance slopes: this provides a one order of magnitude larger sensitivity of phase to refractive index variations; (ii) Under a proper design of a detection scheme, phase noises can be orders of magnitude lower compared to amplitude ones, which results in a much better signal-to-noise ratio; (iii) Phase offers much better possibilities for signal averaging and filtering, as well as for image treatment. Applying a phase-sensitive SPR polarimetry scheme and using gas calibration model, we experimentally demonstrate the detection limit of 10(-8) RIU, which is about two orders of magnitude better compared to amplitude-sensitive schemes. Finally, we show how phase can be employed for filtering and treatment of images in order to improve signal-to-noise ratio even in relatively noisy detection schemes. Combining a much better physical sensitivity and a possibility of imaging and sensing in micro-arrays, phase-sensitive methodologies promise a substantial upgrade of currently available SPR technology. PMID- 19997359 TI - White-emissive tandem-type hybrid organic/polymer diodes with (0.33, 0.33) chromaticity coordinates. AB - This study reports fabrication of white-emissive, tandem-type, hybrid organic/polymer light-emitting diodes (O/PLED). The tandem devices are made by stacking a blue-emissive OLED on a yellow-emissive phenyl-substituted poly(para phenylene vinylene) copolymer-based PLED and applying an organic oxide/Al/molybdenum oxide (MoO(3)) complex structure as a connecting structure or charge-generation layer (CGL). The organic oxide/Al/MoO(3) CGL functions as an effective junction interface for the transport and injection of opposite charge carriers through the stacked configuration. The electroluminescence (EL) spectra of the tandem-type devices can be tuned by varying the intensity of the emission in each emissive component to yield the visible-range spectra from 400 to 750 nm, with Commission Internationale de l'Eclairage chromaticity coordinates of (0.33, 0.33) and a high color rendering capacity as used for illumination. The EL spectra also exhibit good color stability under various bias conditions. The tandem-type device of emission with chromaticity coordinates, (0.30, 0.31), has maximum brightness and luminous efficiency over 25,000 cd/m(2) and approximately 4.2 cd/A, respectively. PMID- 19997360 TI - Efficient sub-Doppler laser cooling of an Indium atomic beam. AB - We have realized efficient transverse cooling of an Indium atomic beam by combining optical pumping with a closed cycle UV laser cooling transition at 325.6 nm. The transverse velocity of the atomic beam is reduced to 13.5+/-3.8 cm/s, well below the Doppler cooling limit. The fraction of laser-cooled In atoms is enhanced to 12+/-3 % by optical pumping in the present experiment. It can be scaled up to approach 100% efficiency in cooling, providing high brightness atomic beams for further applications. Our results establish In on the map of elements suitable for applications involving laser cooling. PMID- 19997361 TI - Strong tunable slow and fast lights using a gain-clamped semiconductor optical amplifier. AB - Previously demonstrated slow light is still far from applications, particularly due to the limited bandwidth and control speed. Although semiconductor-based slow light has the high bandwidth and sub-nanosecond control speed, slow light was observed only in the absorption regime with attenuation, while fast light observed in the gain regime with amplification. The large power difference in two regimes makes the use of the optical delay impractical. We report novel slow light in the gain regime, with a high power comparable to that of fast light, utilizing the anomalous gain characteristic in a gain-clamped semiconductor optical amplifier. The slow light is tunable to fast light with the current as the only variable. Additional high speed operation, fast delay control, and wide range of operation wavelength make the present approach practical. PMID- 19997362 TI - Understanding near/far-field engineering of optical dimer antennas through geometry modification. AB - Numerical investigations based on the boundary element method (BEM) have been carried out to two-dimensional (2-D) silver dimer nano-antennas of various geometries. The near-field and far-field properties are mainly determined by the local geometry at the gap and the global shape of the antenna shafts respectively. A hybrid dimer antenna, which mixes the geometry ingredients of the rod dimer and the bowtie, benefits in both near and far field. Using a microcavity representation, the resonance in dimer nano-antennas is explained in a common and semi-analytical manner. The plasmonic enhancement and the wavelength mismatching in the optical dimer antenna are naturally embodied in this model. The quality factor of the resonance, which can be influenced by the wavelength and the geometry, is discussed intuitively. The understanding presented in this work could guide the future engineering of the optical dimer antenna. PMID- 19997363 TI - Stokes polarimeter optimization in the presence of shot and Gaussian noise. AB - An error minimization method is presented for Stokes polarimeters applicable when the detected signals are affected by a combination of shot and Gaussian noise. The expectation of the Stokes vector variance is used as a performance measure. This measure is compared with the condition number of a polarization state analyzer matrix that is commonly used as a figure of merit. We show that a polarimeter with the minimum condition number is not necessarily optimal. The approach is used to optimize existing prism based polarimeters giving improvements in the performance when shot-noise cannot be neglected. PMID- 19997364 TI - Oblique electron-beam evaporation of distinctive indium-tin-oxide nanorods for enhanced light extraction from InGaN/GaN light emitting diodes. AB - This paper presents a novel and mass-producible technique to fabricate indium-tin oxide (ITO) nanorods which serve as an omnidirectional transparent conductive layer (TCL) for InGaN/GaN light emitting diodes (LEDs). The characteristic nanorods, prepared by oblique electron-beam evaporation in a nitrogen ambient, demonstrate high optical transmittance (T>90%) for the wavelength range of 450nm to 900nm. The light output power of a packaged InGaN/GaN LED with the incorporated nanorod layer is increased by 35.1% at an injection current of 350mA, compared to that of a conventional LED. Calculations based on a finite difference time domain (FDTD) method suggest that the extraction enhancement factor can be further improved by increasing the thickness of the nanorod layer, indicating great potential to enhance the luminous intensity of solid-state lighting devices using ITO nanorod structures. PMID- 19997365 TI - Frequency comb swept lasers. AB - We demonstrate a frequency comb (FC) swept laser and a frequency comb Fourier domain mode locked (FC-FDML) laser for applications in optical coherence tomography (OCT). The fiber-based FC swept lasers operate at a sweep rate of 1kHz and 120kHz, respectively over a 135nm tuning range centered at 1310nm with average output powers of 50mW. A 25GHz free spectral range frequency comb filter in the swept lasers causes the lasers to generate a series of well defined frequency steps. The narrow bandwidth (0.015nm) of the frequency comb filter enables a approximately -1.2dB sensitivity roll off over approximately 3mm range, compared to conventional swept source and FDML lasers which have -10dB and -5dB roll offs, respectively. Measurements at very long ranges are possible with minimal sensitivity loss, however reflections from outside the principal measurement range of 0-3mm appear aliased back into the principal range. In addition, the frequency comb output from the lasers are equally spaced in frequency (linear in k-space). The filtered laser output can be used to self clock the OCT interference signal sampling, enabling direct fast Fourier transformation of the fringe signals, without the need for fringe recalibration procedures. The design and operation principles of FC swept lasers are discussed and designs for short cavity lasers for OCT and interferometric measurement applications are proposed. PMID- 19997366 TI - Spectral analysis of induced color change on periodically nanopatterned silk films. AB - We demonstrate controllable structural color based on periodic nanopatterned 2D lattices in pure protein films of silk fibroin. We show here periodic lattices in silk fibroin films with feature sizes of hundreds of nanometers that exhibit different colors as a function of varying lattice spacing. Further, when varying the index of refraction contrast between the nanopatterned lattice and its surrounding environment by applying liquids on top of the lattices, colorimetric shifts are observed. The effect is characterized experimentally and theoretically and a simple example of glucose concentration sensing is presented. This is the first example of a functional sensor based on silk fibroin optics. PMID- 19997367 TI - Photon energy entanglement characterization by electronic transition interference. AB - We propose photon energy qubits and schemes for photon energy entanglement characterization. Bell inequality violation for energy qubits and complete Bell state analysis are demonstrated theoretically. Photon energy superposition state detection is performed by a two-photon absorption interferometer based on electron transition path interference. The scheme can be realized at room temperature by two-level systems and semiconductor devices. PMID- 19997368 TI - Lamellar grating optimization for miniaturized fourier transform spectrometers. AB - Microfabricated Lamellar grating interferometers (LGI) require fewer components compared to Michelson interferotemeters and offer compact and broadband Fourier transform spectrometers (FTS) with good spectral resolution, high speed and high efficiency. This study presents the fundamental equations that govern the performance and limitations of LGI based FTS systems. Simulations and experiments were conducted to demonstrate and explain the periodic nature of the interferogram envelope due to Talbot image formation. Simulations reveal that the grating period should be chosen large enough to avoid Talbot phase reversal at the expense of mixing of the diffraction orders at the detector. Optimal LGI grating period selection depends on a number of system parameters and requires compromises in spectral resolution and signal-to-bias ratio (SBR) of the interferogram within the spectral range of interest. New analytical equations are derived for spectral resolution and SBR of LGI based FTS systems. PMID- 19997369 TI - Mode expansion and Bragg filtering for a high-fidelity fiber-based photon-pair Source. AB - We report the development of a fiber-based single spatial-mode source of photon pairs where the efficiency of extracting photon-pairs is increased through the use of fiber-end expansion and Bragg filters. This improvement in efficiency enabled a spectrally bright and pure photon-pair source having a small second order correlation function (0.03) and a raw spectral brightness of 44,700 pairs s(-1)nm(-1)mW(-1). The source can be configured to generate entangled photon pairs, characterized via optimal and minimal quantum state tomography, to have a fidelity of 97% and tangle of 92%, without subtracting any background. PMID- 19997370 TI - Propagation-dependent beam profile distortion associated with the Goos-Hanchen shift. AB - The propagation-dependent profile distortion of the reflected beam is studied via deriving the theoretical model of the optical field distribution in both the near and far field. It is shown that strong and fast-varying beam distortions can occur along the propagation path, compared to the profile on the reflecting surface. Numerical simulations for the case of a typical SPR configuration with a sharp angular response curve reveal that, when the phase distribution in the angular range covered by the input beam becomes nonlinear, previous theories based on the linear phase approximation fail to predict the Goos-Hanchen shift and its propagation-dependent variations precisely. Our study could shed light on more accurate modeling of the Goos-Hanchen effect's impact on the relevant photonic devices and measurement applications. PMID- 19997371 TI - Symmetric hybrid surface plasmon polariton waveguides for 3D photonic integration. AB - A two-dimensional symmetric hybrid plasmonic waveguide that integrates two high refractive-index dielectric slabs with a finite-width insulator-metal-insulator (IMI) structure is proposed, and the characteristics of its long-range propagation mode are numerically analyzed at 1550 nm wavelength. In contrast to the previously studied structures, the gap between the slabs and the metal stripe and the associated field enhancement effect result in the dramatically modified modal behavior. It is shown that, under optimized configurations, the transmission loss can be reduced significantly with little change in the mode confinement capability compared to similar dielectric-loaded surface plasmon polariton waveguides. Studies on the crosstalk between adjacent such hybrid waveguides reveal the ability to increase the integration density by approximately 60 times compared with the traditional IMI structures when used in 3D photonic circuits. The studied waveguide could be an interesting alternative to realize high density photonic circuits. PMID- 19997372 TI - Equivalent configurations of optical transformation media. AB - We demonstrate that a medium consisting of two adjoining distinct layers of transformation materials, corresponding respectively to two linear coordinate transformations, can behave effectively as that of the same region transformed by another linear transformation. The equivalence means that, irrespective of the direction of incident wave, the fields of the medium exterior to the transformed regions of the two configurations are exactly the same. This property can also apply to a domain that is transformed by a piecewise linear transformation function, and to a medium that is mapped by a general curved function. This proof is shown analytically based on a rigorous Fourier-Bessel analysis. The equivalence suggests that, for a given transformed domain, one can find an infinite number of complementary media that altogether can give a desired effective response of certain transformation path. PMID- 19997373 TI - Device based in-chip critical dimension and overlay metrology. AB - The use of optical metrology techniques in process control for microelectronic manufacturing has become widespread. These techniques are fast and non destructive, allowing a higher sampling rate than non-optical methods like scanning electron or atomic force microscopy. One drawback of most optical metrology tools is the requirement that special measurement structures be fabricated in the scribe line between chips. This poses significant limitations regarding the characterization of lithography processes that may be overcome via in-chip measurements. In this paper we present experimental results for an in chip optical metrology technique that allows direct measurement of both critical dimensions and overlay displacement errors in the DRAM manufacturing process. This technique does not require special target structures and is performed on the actual semiconductor devices. PMID- 19997374 TI - Emission properties of quantum dots in polymer optical fibres. AB - CdSe/ZnS core-shell quantum dots have been embedded within microstructured polymer optical fibres. The emission properties of quantum dots within fibres have been explored to show that variation in concentration, sample length and pumping regimes effects the emission characteristics of these quantum dots. PMID- 19997375 TI - Experimental investigation on the performance of closely spaced multi-carrier PDM QPSK with digital coherent detection. AB - We experimentally investigate the performance of a spectrally efficient multi carrier channel consisting of two or more optical carriers spaced around the baud rate, with each carrier modulated with polarization division multiplexed (PDM) quadrature phase shift keyed (QPSK) format. We first study the performance of a 100-Gb/s 2-carrier PDM-QPSK channel with each carrier modulated at 12.5 Gbaud as a function of various design parameters such as the time alignment between the modulated carriers, the frequency separation between the carriers, the oversampling factor at the receiver, and the bandwidth of the digital pre-filter used for carrier separation. While the measurements confirm the previously reported observations, they also reveal some interesting additional features. The coherent crosstalk between the modulated carriers is found to be minimized when these carriers are symbol aligned. Spacing the carriers at the baud rate, corresponding to the orthogonal frequency-division multiplexing (OFDM) condition, leads to a local maximum in performance only for some specific cases where large oversampling (>2 x ) is applied. It is found that 4 x oversampling, together with a constant modulus algorithm (CMA) based digital equalizer having multiple quarter-symbol (T/4) spaced taps, gives much better overall performance than 2 x oversampling with a CMA-based equalizer having T/2 spaced taps. In addition, using a T/4-delay-and-add filter (DAF) as a pre-filter for assist carrier separation is found to give better performance than the commonly used T/2-DAF. In addition, it is possible to set the carrier spacing to be as small as 80% of the baud rate while incurring negligible penalty at BER approximately 10(-3). 3 carrier and 5-carrier PDM-QPSK channels at 12.5-Gbaud with frequency-locked carriers spaced at 12.5 GHz and 4 x oversampling are also studied, and shown to perform reasonably well with small relative penalties. Finally, increasing the baud rate of the 2-carrier PDM-QPSK to 25 Gbaud and 28 Gbaud is investigated. It is found that with a fixed sampling speed of 50 Gsamples/s, scaling from 12.5 Gbaud to 25 and 28 Gbaud causes excess crosstalk penalties of about 2.8 dB and 4.8 dB, respectively, indicating the need to increase the sampling speed and transmitter bandwidth in order to support these high-data-rate channels without excessive coherent crosstalk. PMID- 19997376 TI - Organic light-emitting devices fabricated using a premetered coating process. AB - We present the results of a study of flat and uniform organic electroluminescent (EL) layers produced using a simple premetered horizontal-dipping process. It is shown that this process can produce high quality organic semiconductor thin films by utilizing the downstream meniscus of the solution, which may be controlled by adjusting the gap height and the carrying speed. It is also shown that the organic light emitting devices (OLEDs) produced using this method exhibit a peak brightness in excess of 52,000 cd/m(2) and a maximum efficiency of 24 cd/A, with a large active area. From these results, we show that this premetered process for solution coating offers considerable promise for the production of highly efficient, reliable, and large-area solution-processed OLEDs. PMID- 19997377 TI - Highly efficient organic light-emitting devices beyond theoretical prediction under high current density. AB - We develop a simple method to improve external quantum efficiencies (EQEs) of OLEDs under a wide range of current density. An insulating inorganic ultrathin layer (LiF) was sandwiched between exciton formation layer and electron transporting layer. A maximal EQE of 5.9% in a DCM based fluorescent OLED, which far exceeds the theoretical upper limit of 3.7%, was obtained under the current density of 487 mA/cm(2) with a brightness maximum of 76740 cd/m(2). The similar electroluminescence properties were also obtained in a C545T based green OLED using this method. The overall enhancement of EQE, and the nonlinear enhancement of EQE at high current density in these devices are attributed to the effect of electrical field on excitons. PMID- 19997378 TI - Analysis of fast full angle fluorescence diffuse optical tomography with beam forming illumination. AB - Challenges remain in imaging fast biological activities through whole body using fluorescence diffuse optical tomography (FDOT). We propose and analyze three full angle FDOT systems with different beam-forming illuminations (BF-FDOT), including line illumination (L-FDOT), area illumination (A-FDOT), and multiple-points illumination (MP-FDOT). Singular value analysis and experimental validation are used to optimize the experimental parameters in terms of hardware design, data collection and utilization. Comparisons are made on the system performance between L-FDOT and the conventional point illumination based full angle FDOT system (P-FDOT) with both numerical simulation and phantom experiment. We demonstrate that at least three cycles of projections are needed for P-FDOT to achieve comparable whole body image quality with L-FDOT. We also compare these three BF-FDOT systems and further discuss how these optimized parameters can be employed to improve spatial and temporal performances within current computational capacities, and guide the design of the BF-FDOT systems. PMID- 19997379 TI - Rapid Mueller matrix polarimetry based on parallelized polarization state generation and detection. AB - We present rapid Mueller matrix polarimetry that can extract twelve Muller matrix elements from a single intensity image in real time and with high spatial resolution. This is achieved by parallelizing the respective polarization state generation and polarization state detection processes, which in existing polarimeters is performed sequentially. Parallelization of the polarization state generation process is accomplished through the use of vector beams, for which this work represents a new application domain. Polarization state detection is parallelized by uniquely combining a microscope/array detector setup with a specialized algorithm that simultaneously utilizes information from multiple spatial regions of the array detector. Simulated results applying this technique to two anisotropic samples including metamaterial yield material parameters that are consistent with those reported in the literature. PMID- 19997380 TI - Color temperature tunable white light emitting diodes packaged with an omni directional reflector. AB - This study proposed a correlated color temperature (CCT) tunable phosphor converted white light emitting diode (LED) with an omni-directional reflector (ODR). Applying current to each individual InGaN based ultraviolet, purple and blue source LED chip of the white LED package, we can achieve the CCT tunability. The optimum color properties of the resulting white light are (0.3347, 0.3384), 5398 K, 81, 3137-8746 K for color coordinates, CCT, color rendering index (CRI) and CCT tuning range, respectively. Roughening the ODR substrate, we solve the non-uniformity color distribution caused by the reflectance of the ODR and positioning of source LED chips. PMID- 19997381 TI - Multispectral optoacoustic tomography (MSOT) scanner for whole-body small animal imaging. AB - A major difficulty arising from whole-body optoacoustic imaging is the long acquisition times associated with recording signals from multiple spatial projections. The acquired signals are also generally weak and the signal-to-noise ratio is low, problems often solved by signal averaging, which complicates acquisition and increases acquisition times to an extent that makes many in vivo applications challenging or even impossible. Herein we present a fast acquisition multispectral optoacoustic tomography (MSOT) scanner for whole-body visualization of molecular markers in small animals. Multi-wavelength illumination offers the possibility to resolve exogenously administered fluorescent probes, biomarkers, and other intrinsic and exogenous chromophores. The system performance is determined in phantom experiments involving molecular probes and validated by imaging of small animals of various scales. PMID- 19997382 TI - Non-Gaussian modes in a HeNe laser. AB - Single transverse mode oscillation is realized in a conventional HeNe laser outside the stability region of the optical resonator. Depending on the mirror separation different spatial modes can be generated. The mode volume of these modes is laterally limited by the diameter of the discharge capillary rather than by the beam waist of a stable Gaussian mode. Numerical solution of the Maxwell equations with appropriate boundary conditions shows good agreement with the observations. Such modes could potentially facilitate single transverse mode operation of waveguide lasers and fiber lasers. PMID- 19997383 TI - Temperature-dependent Goos-Hanchen shift on the interface of metal/dielectric composites. AB - The temperature-dependent Goos-Hanchen shift (GHS) for an electromagnetic wave reflected from a metal/dielectric composite material is investigated. With the stationary-phase method, we theoretically show that the effect of the temperature on GHS is significant near the Brewster angle for the dielectric composites and at the grazing angle for the metallic composites. For dielectric composites, the lateral shift can be negative as well as positive. And GHS may become much negative, much positive, and nonmonotonic variation with increasing the temperature under different conditions. Moreover, through the suitable adjustment of the temperature, one may realize the reversal of the GHS. To support the above results, numerical simulations for Gaussian incident beams based on the momentum method and COMSOL Multiphysics software are provided, and reasonable agreement between the theoretical results and numerical simulations is found. PMID- 19997384 TI - Time delays in the synchronization of chaotic coupled lasers with feedback. AB - Isochrony and time leadership was studied in the synchronized excitable behavior of coupled chaotic diode lasers. Each unit of the system had chaos due to feedback with a fixed delay time. The inter-units coupling signal had a second, independent, characteristic time. Synchronized excitable spikes present isochronous, time leading or time lagging behavior whose stability is shown to depend on a simple relation between the feedback and the coupling times. Experiments on the synchronized low frequency fluctuations of two optically coupled semiconductor lasers and numerical calculations with coupled laser equations verify the predicted stability conditions for synchronization. Synchronism with intermittent time leadership exchange was also observed and characterized. PMID- 19997386 TI - Delay control in attosecond pump-probe experiments. AB - The time delay between the pump and probe pulses in attosecond time-resolved experiments, such as attosecond streaking, is commonly introduced by splitting and recombining the two pulses in an interferometer. This technique suffers from instability in the optical path lengths of the two arms due to mechanical vibration of the optical elements and fluctuating environmental conditions. We present a technique with which the instability of the unconventional interferometer is suppressed while at the same time the time delay is controlled to within 20 as RMS using a feedback loop. Using this scheme, the streaked spectrogram of an attosecond pulse was measured. PMID- 19997385 TI - Semiconducting polymer waveguides for end-fired ultra-fast optical amplifiers. AB - A method to fabricate conjugated polymer waveguides with well defined edge facets is demonstrated. The utility of the approach is explored for application as end fired ultrafast optical amplifiers based on poly(9,9'-dioctylfluorene-co benzothiadiazole). An internal gain of 19 dB was achieved on a 760 microm long waveguide at 565 nm wavelength. This fabrication procedure may be applied to a wide range of conjugated polymers and organic light-emitting devices, providing an important step towards future applications of organic integrated photonics. PMID- 19997387 TI - Diffraction engineering of multimode waveguides using computer-generated planar holograms. AB - The self-imaging property in multimode waveguides is related to the waveguide widths and lengths. By engineering the diffraction properties of multimode waveguides, we propose a scheme to design devices with reduced self-imaging lengths at a fixed width. Using computer-generated planar holograms, the coupling coefficients between the guided modes are adjusted to generate the desired diffraction properties. Calculations based on the coupled-mode theory are presented. Devices are designed based on a silicon-on-insulator (SOI) platform. Beam propagation simulations are used to verify the coupled-mode theory analysis. PMID- 19997388 TI - Radiation force of scalar and electromagnetic twisted Gaussian Schell-model beams. AB - Radiation force of a focused scalar twisted Gaussian Schell-model (TGSM) beam on a Rayleigh dielectric sphere is investigated. It is found that the twist phase affects the radiation force and by raising the absolute value of the twist factor it is possible to increase both transverse and longitudinal trapping ranges at the real focus where the maximum on-axis intensity is located. Numerical calculations of radiation forces induced by a focused electromagnetic TGSM beam on a Rayleigh dielectric sphere are carried out. It is found that radiation force is closely related to the twist phase, degree of polarization and correlation factors of the initial beam. The trapping stability is also discussed. PMID- 19997389 TI - Stable UV to IR supercontinuum generation in calcium fluoride with conserved circular polarization states. AB - The supercontinuum generated with a linearly polarized near-IR (775 nm) pump in rotated calcium fluoride is shown to have intrinsic intensity and polarization modulations. To mask the rotation of the crystal plate, we circularly polarize the pump and find greatly improved output parameters for the generated white light: intensity fluctuations of 0.5% limited only by pump laser stability, and a circular polarization state-matching that of the pump-over the entire visible spectrum. This polarization conservation allows the return of the supercontinuum to a linear polarization state or to a pair of linearly polarized beams with correlated intensity fluctuations. We were also able to extend the supercontinuum source deep into the ultraviolet with a frequency doubled (387 nm) pump, to serve as an new source to probe the region where most molecular photochemistry occurs. PMID- 19997390 TI - Modulation instability, Akhmediev Breathers and continuous wave supercontinuum generation. AB - Numerical simulations of the onset phase of continuous wave supercontinuum generation from modulation instability show that the structure of the field as it develops can be interpreted in terms of the properties of Akhmediev Breathers. Numerical and analytical results are compared with experimental measurements of spectral broadening in photonic crystal fiber using nanosecond pulses. PMID- 19997391 TI - Pump power dependence of Kerr signals in femtosecond cross pump-probe optical Kerr measurements. AB - We investigated the influence of self-diffraction effect on femtosecond cross pump-probe optical Kerr shutter (OKS) measurements for fast response and slow response materials, respectively. In the slow response material of CS(2), a fast response resulted from self-diffraction effect was observed in the time-resolved OKS signals, and the signal intensity showed different pump power dependences at different delay time. For the fast response material of bismuth oxide (BI) glass, the pump power dependence of OKS signals varied when the polarization angle was varied. The pump power dependences of OKS signals for the both materials showed that, the OKS signals resulted mainly from laser induced birefringence effect at low pump powers, and the contribution of self-diffraction effect to OKS signals was enhanced with increasing the pump power. PMID- 19997392 TI - Counterpropagating surface solitons in two-dimensional photonic lattices. AB - We study interaction of counterpropagating beams in truncated two-dimensional photonic lattices induced optically in photorefractive crystals, and demonstrate the existence of counterpropagating surface solitons localized in the lattice corners and at the edges. We display intriguing dynamical properties of such composite optical beams and reveal that the lattice surface provides a strong stabilization effect on the beam propagation. We also observe dynamical instabilities for stronger coupling and longer propagation distances in the form of beam splitting. No such instabilities exist in the single beam surface propagation. PMID- 19997393 TI - Hybrid surface-enhanced Raman scattering substrate from gold nanoparticle and photonic crystal: maneuverability and uniformity of Raman spectra. AB - A novel hybrid surface-enhanced Raman scattering (SERS) substrate based on Au nanoparticles decorated inverse opal (IO) photonic crystal (PhC) is presented. In addition to the enhancement contributed from Au nanoparticles, a desired Raman signal can be selectively further enhanced by appropriately overlapping the center of photonic bandgap of the IO PhC with the wavelength of the Raman signal. Furthermore, the lattice structure of the IO PhC provides excellent control of the distribution of Au nanoparticles to produce SERS spectra with high uniformity. The new design of SERS substrate provides extra maneuverability for ultra-high sensitivity sensor applications. PMID- 19997394 TI - Fabrication of optical mosaic gratings with phase and attitude adjustments employing latent fringes and a red-wavelength dual-beam interferometer. AB - We present a method to make optical mosaic gratings that uses the exposure beams and the latent grating created by the previous exposure to adjust the lateral position and readjust the attitude of the substrate for the current exposure. As thus, it is a direct method without using any auxiliary reference grating(s) and it avoids the asynchronous drifts between otherwise independent exposure and alignment optical sub-systems. In addition, the method uses a red laser wavelength in the plane-mirror interferometers for the multi-dimensional attitude adjustment, so the adjustment can be done at leisure. The mosaic procedure is described step by step, and the principles to minimize substrate alignment errors are explained in detail. Experimentally we made several mosaics of (50 + 30) x 50 mm(2) final grating area. The typical peak-valley and root-mean-square values of the measured -1st-order diffraction wavefront errors are 0.036 lambda and 0.006 lambda, respectively. PMID- 19997395 TI - Efficient second harmonic generation of double-end diffusion-bonded Nd:YVO4 self Raman laser producing 7.9 W yellow light. AB - A high power and efficient 588 nm yellow light is demonstrated through intracavity frequency doubling of an acousto-optic Q-switched self-frequency Raman laser. A 30-mm-length double-end diffusion-bonded Nd:YVO(4) crystal was utilized for efficient self-Raman laser operation by reducing the thermal effects and increasing the interaction length for the stimulated Raman scattering. A 15 mm-length LBO with non-critical phase matching (theta = 90 degrees, phi = 0 degrees) cut was adopted for efficient second-harmonic generation. The focus position of incident pump light and both the repetition rate and the duty cycle of the Q-switch have been optimized. At a repetition rate of 110 kHz and a duty cycle of 5%, the average power of 588 nm light is up to 7.93 W while the incident pump power is 26.5 W, corresponding to an overall diode-yellow conversion efficiency of 30% and a slope efficiency of 43%. PMID- 19997396 TI - Thermally stabilized PCF-based sensor for temperature measurements up to 1000 degrees C. AB - We report on the development of a stable Photonic Crystal Fiber (PCF) based two mode interferometric sensor for ultra-high temperature measurements (up to 1000 degrees C). The device consists of a stub of PCF spliced to standard optical fiber. In the splice regions, the voids of the PCF are fully collapsed, thus allowing the excitation and recombination of two core modes. The device spectrum exhibits sinusoidal interference pattern which shifts with temperature. We show that, despite being compact and robust, the proposed sensor head needs a quite long burn in (thermal annealing) to achieve an adequate and stable functionality level. The burn in process eliminates the residual stress in the fiber structure, which had been accumulated during the drawing phase, and changes the glass fictive temperature. PMID- 19997397 TI - Breaking the feature sizes down to sub-22 nm by plasmonic interference lithography using dielectric-metal multilayer. AB - We have developed the plasmonic interference lithography technique to achieve the feature sizes theoretically down to sub-22 nm even to 16.5 nm by using dielectric metal multilayer (DMM) with diffraction-limited masks at the wavelength of 193 nm with p-polarization. An 8 pairs of GaN (10 nm)/Al (12 nm) multilayer is designed as a filter allowing only a part of high wavevector k (evanescent waves) to pass through for interference lithography. The analysis of the influence by the number of DMM layers is presented. 4 pairs of the proposed multilayer can be competent for pattern the minimal feature size down to 21.5 nm at the visibility about 0.4 to satisfy the minimum visibility required with positive resist. Finite difference time-domain analysis method is used to demonstrate the validity of the theory. PMID- 19997398 TI - Simultaneous remote extraction of multiple speech sources and heart beats from secondary speckles pattern. AB - The ability of dynamic extraction of remote sounds is very appealing. In this manuscript we propose an optical approach allowing the extraction and the separation of remote sound sources. The approach is very modular and it does not apply any constraints regarding the relative position of the sound sources and the detection device. The optical setup doing the detection is very simple and versatile. The principle is to observe the movement of the secondary speckle patterns that are generated on top of the target when it is illuminated by a spot of laser beam. Proper adaption of the imaging optics allows following the temporal trajectories of those speckles and extracting the sound signals out of the processed trajectory. Various sound sources are imaged in different spatial pixels and thus blind source separation becomes a very simple task. PMID- 19997399 TI - One-step fabrication of nanostructures by femtosecond laser for surface-enhanced Raman scattering. AB - This paper reports an efficient fabrication of nanostructures on silicon substrates for surface-enhanced Raman scattering (SERS). Silicon wafer substrates in the aqueous solution of silver nitrate were machined by the femtosecond laser direct writing to achieve simultaneously in one-step the generation of grating like nanostructures on the surface of the substrate and the formation of silver nanoparticles on the surface of the nanostructures via the laser-induced photoreduction effect. Parametric studies were conducted for the different concentrations of aqueous silver nitrate solutions and scanning speeds. The enhancement factor of the SERS is found to be higher than 10(9). The patterning technique provides an opportunity to incorporate the SERS capability in a functional microchip. PMID- 19997400 TI - Spherically shaped micro-structured antireflective surfaces. AB - An antireflecting micro-structured interface, working in the resonance domain, and made from a bi-periodic array of semi-spherical hollowing-out in a silicon substrate is presented. Its parameters such as sphere radius and position of sphere centers from the surface are optimized numerically. A simple and robust process is described allowing such kind of antireflective surfaces to be fabricated for the infrared range. Spectral and angular reflectance measurement demonstrates the efficiency of the antireflective micro-structured interface which can easily be adapted for the visible range and for photovoltaic applications by a simple homothetic modification of the micro-structure typical dimensions. PMID- 19997401 TI - Passive interferometric array optimization based on redundant spacing calibration. AB - An optimization of passive interferometric circular arrays for redundant spacing calibration (RSC) is advanced to eliminate phase errors of the array system. The principle of RSC is presented to solve corresponded constraints for passive interferometric circular arrays. The simulated annealing algorithm (SAA) is introduced to settle the array optimization with a criterion of maximizing the distance between u-v points. The optimized circular arrays with element numbers of 8 to 16 antennas are laid out, and RSC is used for the optimized ten-element passive interferometric circular array. PMID- 19997402 TI - Fabrication and characterization of a chalcogenide-tellurite composite microstructure fiber with high nonlinearity. AB - A highly nonlinear composite fiber, which has a 1.5 microm chalcogenide glass core surrounded by a tellurite glass microstructure cladding, has been fabricated by the method of stack and draw. A tellurite glass capillary containing a As(2)S(3) rod was sealed with negative pressure inside. Then this capillary and other empty capillaries were stacked into a tellurite glass tube, and elongated into a cane. This cane was then inserted into another tellurite glass jacket tube and drawn into the composite microstructure fiber. The fiber has a flattened chromatic dispersion together with a zero dispersion wavelength located in the near infrared range. The propagation losses at 1.55 microm were 18.3 dB/m. The nonlinear coefficient at 1.55 microm was 9.3 m(-1)W(-1). Such a high nonlinear coefficient counteracts the high propagation losses to a large extent. A supercontinuum spectrum of 20-dB bandwidth covering 800-2400 nm was generated by this composite microstructure fiber. PMID- 19997403 TI - Spatial and polarization entanglement of lasing patterns and related dynamic behaviors in laser-diode-pumped solid-state lasers. AB - To provide the underlying physical mechanism for formations of spatial- and polarization-entangled lasing patterns (namely, SPEPs), we performed experiments using a c-cut Nd:GdVO(4) microchip laser with off-axis laser-diode pumping. This extends recent work on entangled lasing pattern generation from an isotropic laser, where such a pattern was explained only in terms of generalized coherent states (GCSs) formed by mathematical manipulation. Here, we show that polarization-resolved transverse patterns can be well explained by the transverse mode-locking of distinct orthogonal linearly polarized Ince-Gauss (IG) mode pairs rather than GCSs. Dynamic properties of SPEPs were experimentally examined in both free-running and modulated conditions to identify long-term correlations of IG mode pairs over time. The complete chaos synchronization among IG mode pairs subjected to external perturbation is also demonstrated. PMID- 19997404 TI - A self-Q-switched all-fiber erbium laser at 1530 nm using an auxiliary 1570-nm erbium laser. AB - We demonstrate a self-Q-switched, all-fiber, tunable, erbium laser at 1530 nm with high pulse repetition rates of 0.9-10 kHz. Through the use of an auxiliary 10-mW, 1570 nm laser that shortened the relaxation time of erbium, sequentially Q switched pulses with pulse energies between 4 and 6 microJ and pulse widths of 40 ns were steadily achieved. A peak pulse power of 165 W was obtained. PMID- 19997406 TI - Laser-induced damage of KDP crystals by 1omega nanosecond pulses: influence of crystal orientation. AB - We investigate the influence of THG-cut KDP crystal orientation on laser damage at 1064 nm under nanosecond pulses. Since laser damage is now assumed to initiate on precursor defects, this study makes a connection between these nanodefects (throughout a mesoscopic description) and the influence of their orientation on laser damage. Some investigations have already been carried out in various crystals and particularly for KDP, indicating propagation direction and polarization dependences. We performed experiments for two orthogonal positions of the crystal and results clearly indicate that KDP crystal laser damage depends on its orientation. We carried out further investigations on the effect of the polarization orientation, by rotating the crystal around the propagation axis. We then obtained the evolution of the damage probability as a function of the rotation angle. To account for these experimental res ts, we propose a laser damage model based on ellipsoid-shaped defects. This modeling is a refined implementation of the DMT model (Drude Mie Thermal) [Dyan et al., J. Opt. Soc. Am. B 25, 1087-1095 (2008)], by introducing absorption efficiency calculations for an ellipsoidal geometry. Modeling simulations are in good agreement with experimental results. PMID- 19997405 TI - Retinal imaging with polarization-sensitive optical coherence tomography and adaptive optics. AB - Various layers of the retina are well known to alter the polarization state of light. Such changes in polarization may be a sensitive indicator of tissue structure and function, and as such have gained increased clinical attention. Here we demonstrate a polarization-sensitive optical coherence tomography (PS OCT) system that incorporates adaptive optics (AO) in the sample arm and a single line scan camera in the detection arm. We quantify the benefit of AO for PS-OCT in terms of signal-to-noise, lateral resolution, and speckle size. Double pass phase retardation per unit depth values ranging from 0.25 degrees/microm to 0.65 degrees/microm were found in the birefringent nerve fiber layer at 6 degrees eccentricity, superior to the fovea, with the highest values being noticeably higher than previously reported with PS-OCT around the optic nerve head. Moreover, fast axis orientation and degree of polarization uniformity measurements made with AO-PS-OCT demonstrate polarization scrambling in the retinal pigment epithelium at the highest resolution reported to date. PMID- 19997407 TI - Optical gain and penalty characteristics of a fiber-Bragg-grating based active optical-add-drop multiplexer. AB - We investigate optical gain and penalty characteristics of a fiber-Bragg-grating based active optical-add-drop multiplexer (OADM). The active OADM has a two-stage structure and can amplify all the add/drop/thru channels simultaneously. The drop channel penalty is a key parameter for the reliable operation of the active OADM. This penalty, caused mainly by multiple reflections within the active OADM, can be minimized adjusting pump powers. At the minimum drop-channel penalty, our active OADM shows fairly high optical gains as 15.5, 21, and 19.4 dB for the add, drop, and thru channels, respectively. Nevertheless, the penalties of these channels are comparable to the passive OADM. PMID- 19997408 TI - Tuning of an active photonic crystal cavity by an hybrid silica/silicon near field probe. AB - The influence of a near-field tip on the spectral characteristics of a resonant mode of an active photonic crystal micro-cavity was investigated. The wavelength shift of the mode was theoretically and experimentally demonstrated and evaluated as a function of the nature and the position of the tip above the cavity. Experiment showed that the shift induced is ten times higher with a Si-coated silica probe than with a bare silica tip: a shift until 2 nm was reached with Si coated tip whereas the shift with bare silica tip is in the range of the tenth of nanometer, for wavelengths around 1,55 microm. PMID- 19997409 TI - Multiple traps created with an inclined dual-fiber system. AB - Multiple optical traps allow one to manipulate multiple particles simultaneously, to characterize interactions in colloidal systems, and to assemble particles into complex structures. Most of the current multiple optical traps are realized with microscope objective-based optical tweezers, which are bulky in size. In this article, we created multiple optical traps with an inclined dual-fiber optical tweezers setup. One 3D trap and two 2D traps were formed at different vertical levels with adjustable separations and positions. We demonstrated that this fiber based trapping system can be used as a simple block to perform multiple functions, such as particle grouping, separation, and stacking. Moreover, we found that multiple beads can be trapped and stacked up in three dimensions. Compared with those formed with objective-based optical tweezers, the multiple traps presented here are small in size and independent of the objective or the substrate, and hence hold the promise to be integrated in microfluidic systems. This fiber-based multiple traps can be used for on-chip parallel manipulation, particle separation, and characterization of interactions of colloidal and biological systems. PMID- 19997410 TI - High efficient actively Q-switched Ho:LuAG laser. AB - We present the room temperature Q-switched performances of a Ho:LuAG laser operated at 2.1 microm. At the repetition rate of 10 kHz, the maximum average output power of 9.9 W with a slope efficiency of 69.9% relative to absorbed pump power was obtained in Ho:LuAG laser. Also, the minimum pulse width of 33.0 ns was obtained, corresponding to the peak power was 30.0 kW. PMID- 19997411 TI - Organic dye lasers with distributed Bragg reflector grating and distributed feedback resonator. AB - The paper presents polymeric waveguide dye laser with distributed Bragg reflector (DBR) grating or distributed feedback (DFB) resonator. DBR grating and DFB resonator were fabricated on a surface of SU-8 2002 photoresist polymer using interference of laser beams. Polystyrene (PS) and poly(vinyl butyral) (PVB) waveguides dispersed with laser dye of 3-(2-benzothiazolyl)-7-(diethylamino) coumarin (Coumarin 6) and tris(8-quinolinolate)aluminum (Alq) as a host were used. Single mode of lasing with TE mode was measured from the polymeric waveguide with DBR grating and DFB resonator. Threshold of lasing with DBR grating is lower than that with DFB resonator. For PVB/Alq/Coumarin 6 waveguide, 0.1 mJ cm(-2) pulse(-1) of threshold was measured with DBR grating and 0.3 mJ cm( 2) pulse(-1) with DFB resonator. Slope efficiency between 0.06 and 0.09 % was measured for PS/Alq/Coumarin 6 waveguide and that between 0.07 and 0.15 for PVB /Alq/Coumarin 6 waveguide. PMID- 19997412 TI - Ultra-sensitive fluorescence spectroscopy of isolated surface-adsorbed molecules using an optical nanofiber. AB - The strong radial confinement and the pronounced evanescent field of the guided light in optical nanofibers yield favorable conditions for ultra-sensitive surface spectroscopy of molecules deposited on the fiber. Using the guided mode of the nanofiber for both excitation and fluorescence collection, we present spectroscopic measurements on 3,4,9,10-perylenetetracarboxylic dianhydride molecules (PTCDA) at ambient conditions. Surface coverages as small as 1 per thousand of a compact monolayer still give rise to fluorescence spectra with a good signal to noise ratio. Moreover, we analyze and quantify the self-absorption effects due to reabsorption of the emitted fluorescence light by circumjacent surface-adsorbed molecules distributed along the fiber waist. PMID- 19997413 TI - Direct observation of lateral photovoltaic effect in nano-metal-films. AB - Lateral photovoltaic effect (LPE) observed on the metal films is unusual because it violates a principle that the LPEs are always observed on the surface of a semiconductor. Compared with early studies, we have realized an obvious metal film LPE in a metal-semiconductor (MS) structure. By further arguing with experimental results, this work also intensively elucidates many features of LPE which the early models never touched upon. All the data and analyses in this study indicate that metal side LPE in MS structure has some natural superiorities to the semiconductor side LPE and may open many exciting opportunities for realizing multifunctional devices. PMID- 19997414 TI - Diffractionless beam in free space with adiabatic changing refractive index in a single mode tapered slab waveguide. AB - We propose a novel design to produce a free space diffractionless beam by adiabatically reducing the difference of the refractive index between the core and the cladding regions of a single mode tapered slab waveguide. To ensure only one propagating eigenmode in the adiabatic transition, the correlation of the waveguide core width and the refractive index is investigated. Under the adiabatic condition, we demonstrate that our waveguide can emit a diffractionless beam in free space up to 500 micrometers maintaining 72% of its original peak intensity. The proposed waveguide could find excellent applications for imaging purposes where an extended depth of field is required. PMID- 19997415 TI - Self-focusing and spatial plasmon-polariton solitons. AB - We study nonlinear propagation of surface plasmon polaritons along an interface between metal and nonlinear Kerr dielectric. We demonstrate numerically self focusing of a plasmon beam at large powers and the formation of slowly decaying spatial soliton in the presence of losses. We develop an analytical model for describing the evolution of spatial plasmon-solitons and observe a good agreement with numerical results. PMID- 19997416 TI - Multi-sample parallel estimation in volume holographic correlator for remote sensing image recognition. AB - Based on volume holographic correlator, a multi-sample parallel estimation method is proposed to implement remote sensing image recognition with high accuracy. The essential steps of the method including image preprocessing, estimation curves fitting, template images preparation and estimation equation establishing are discussed in detail. The experimental results show the validity of the multi sample parallel estimation method, and the recognition accuracy is improved by increasing the sample numbers. PMID- 19997417 TI - Adjustable-chirp VCSEL-to-VCSEL injection locking for 10-Gb/s transmission at 1.55 microm. AB - 1.55-microm single-mode VCSEL frequency chirp behavior is investigated in under threshold and above-threshold operating conditions for different VCSEL-to-VCSEL injection locking configurations with respect to free-running case. We experimentally evaluated the capability of adjusting the frequency chirp, reducing its value and inverting the sign. The control over the frequency chirp is obtained changing the wavelength detuning and power injection ratio between the VCSEL master and the VCSEL slave. Advantages of the chirp inversion are demonstrated for 10 Gb/s error-free propagation at 1.55-microm over 40-km standard single mode fiber without any dispersion compensation. PMID- 19997418 TI - Dependence of dynamic Lorentz frequency shift on carrier-envelope phase and including local field effects. AB - We investigate the local field effects in a ZnO dense medium. Our results show due to the local-field effects, the Lorentz shifts can be found in the reflected spectra driven by the few-cycle laser pulse. Moreover, the dynamic Lorentz shifts depend sensitively on the carrier-envelope phase (CEP) of the few-cycle laser pulse, which provides a useful means to obtain the CEP information by the frequency shifts. PMID- 19997419 TI - In vivo dynamic optical coherence elastography using a ring actuator. AB - We present a novel sample arm arrangement for dynamic optical coherence elastography based on excitation by a ring actuator. The actuator enables coincident excitation and imaging to be performed on a sample, facilitating in vivo operation. Sub-micrometer vibrations in the audio frequency range were coupled to samples that were imaged using optical coherence tomography. The resulting vibration amplitude and microstrain maps are presented for bilayer silicone phantoms and multiple skin sites on a human subject. Contrast based on the differing elastic properties is shown, notably between the epidermis and dermis. The results constitute the first demonstration of a practical means of performing in vivo dynamic optical coherence elastography on a human subject. PMID- 19997420 TI - A tunable and switchable single-longitudinal-mode dual-wavelength fiber laser with a simple linear cavity. AB - A simple linear cavity erbium-doped fiber laser based on a Fabry-Perot filter which consists of a pair of fiber Bragg gratings is proposed for tunable and switchable single-longitudinal-mode dual-wavelength operation. The single longitudinal-mode is obtained by the saturable absorption of an unpumed erbium doped fiber together with a narrow-band fiber Bragg grating. Under the high pump power (>166 mW) condition, the stable dual-wavelength oscillation with uniform amplitude can be realized by carefully adjusting the polarization controller in the cavity. Wavelength selection and switching are achieved by tuning the narrow band fiber Bragg grating in the system. The spacing of the dual-wavelength can be selected at 0.20 nm (approximately 25.62 GHz), 0.22 nm (approximately 28.19 GHz) and 0.54 nm (approximately 69.19 GHz). PMID- 19997421 TI - InP/AlGaInP quantum dot semiconductor disk lasers for CW TEM00 emission at 716 - 755 nm. AB - Multiple layers of InP QDs, self-assembled during epitaxial growth, were incorporated into the active region of an (Al(x)Ga(1-x))(0.51)In(0.49)P based semiconductor disk laser with monolithic Al(x)Ga(1-x)As distributed Bragg reflector. Three gain structure samples were selected from the epitaxial wafer, bonded to single-crystal diamond heatspreaders and optically pumped at 532 nm within a high finesse external laser cavity. Laser emission with peak wavelengths at 716, 729, and 739 nm, respectively, was achieved from the three samples; the latter demonstrating tuning from 729 to 755 nm. Maximum continuous wave output power of 52 mW at 739 nm was achieved with 0.2% output coupling; the threshold and slope efficiency were 220 mW and 5.7% respectively. PMID- 19997422 TI - Femtosecond mode-locked fiber laser employing a hollow optical fiber filled with carbon nanotube dispersion as saturable absorber. AB - We propose a novel in-line saturable absorber incorporating a hollow optical fiber (HOF) filled with single-walled carbon nanotube (SWCNT) dispersion. The evanescent field of the propagating light in the ring core interacts with the SWCNT/polymer composite distributed over the whole length of the HOF. The proposed saturable absorber with all-fiber format offers the robust and long nonlinear interaction along the waveguide direction expecting the increase of the threshold for optical and thermal damages with simple fabrication process. Low concentration SWCNT/polymer composite exhibiting very broadband resonant absorption around 1.5 microm with low scattering loss is prepared and based on this, we successfully demonstrate the passively mode-locked fiber laser including the SWCNT-filled HOF where the spectral bandwidth and the pulse duration of the laser output are 5.5 nm and 490 fs, respectively, with a repetition rate of 18.5 MHz. PMID- 19997423 TI - Mid-infrared near-field spectroscopy. AB - We demonstrate continuous infrared spectra from 20 nm sample spots, by combining dispersive Fourier-transform infrared spectroscopy (FTIR) with scattering near field microscopy (s-SNOM). With the "apertureless" tip of a standard AFM cantilever in one arm of a Michelson interferometer the spectra arise simultaneously in amplitude and phase. The effect of near-field phonon resonance of SiC is used to verify background-free s-SNOM operation, and to determine the absolute scattering efficiency, at 6 cm(-1) spectral resolution. We further report first evidence of free-induction decay from a scatterer composed of parts coupled by near-fields. This is possible only with broadband illumination. It offers a new, unique tool to discriminate against background scattering artifacts. PMID- 19997424 TI - Near-infrared optical response of thin film pH-sensitive hydrogel coated on a gold nanocrescent array. AB - A hydrogel-based chemiresponsive sensor for monitoring H(+) (pH) has been developed by coating the surface of a gold nanocrescent array structure with a thin film of a poly(2-hydroxylethyl methacrylate)-based (poly-HEMA) hydrogel. The transmission measurement results of the close-packed gold nanocrescent array fabricated via electron beam lithography demonstrate near-infrared localized surface plasmon resonance peaks with sensitivities up to 332 nm/RIU in detecting refractive index change. Measurements of the hydrogel under solutions of increasing pH show the plasmon peak blueshifts by 17 nm and the integrated transmission increases by 1.8 in the operating range of 4.5 - 6.4 pH, which is ideal for biochemical sensor applications. PMID- 19997425 TI - ZnO subwavelength wires for fast-response mid-infrared detection. AB - Room temperature operating thermal detection for mid-infrared light based on ZnO subwavelength wires has been demonstrated. Electric resistance in ZnO wires increases linearly with the intensity of incident light. Noise equivalent power (NEP) of 5.8 microW/Hz(1/2) (at 1 kHz) with typical response times as fast as 1.3 ms is obtained at 10.6-microm wavelength. The sensitivity and response time of the detector are also found to be insensitive to the ambient. PMID- 19997426 TI - Nanowires/microfiber hybrid structure multicolor laser. AB - We demonstrate a compact hybrid structure red-green-ultraviolet three-color laser consisting of three distinct semiconductor nanowires (CdSe, CdS and ZnO) attached to a silica microfiber, which is pumped by 355 nm wavelength laser pulses. The exciting of the nanowires and the collection of the photoluminescence (PL) are implemented by means of evanescent coupling through the same silica microfiber. When pump energy higher than 1.3 microJ, three spatially and spectrally distinct lasing groups can be measured at the same output port simultaneously. The approach can be extended to other materials to produce hybrid lasers that cover ultraviolet to near infrared spectral regions. PMID- 19997427 TI - Geometrically robust image watermarking by sector-shaped partitioning of geometric-invariant regions. AB - In a feature-based geometrically robust watermarking system, it is a challenging task to detect geometric-invariant regions (GIRs) which can survive a broad range of image processing operations. Instead of commonly used Harris detector or Mexican hat wavelet method, a more robust corner detector named multi-scale curvature product (MSCP) is adopted to extract salient features in this paper. Based on such features, disk-like GIRs are found, which consists of three steps. First, robust edge contours are extracted. Then, MSCP is utilized to detect the centers for GIRs. Third, the characteristic scale selection is performed to calculate the radius of each GIR. A novel sector-shaped partitioning method for the GIRs is designed, which can divide a GIR into several sector discs with the help of the most important corner (MIC). The watermark message is then embedded bit by bit in each sector by using Quantization Index Modulation (QIM). The GIRs and the divided sector discs are invariant to geometric transforms, so the watermarking method inherently has high robustness against geometric attacks. Experimental results show that the scheme has a better robustness against various image processing operations including common processing attacks, affine transforms, cropping, and random bending attack (RBA) than the previous approaches. PMID- 19997428 TI - LD bar corner-pumped TEM00 CW composite Nd:YAG laser. AB - Ray-tracing algorithm was used to simulate the pump absorption efficiency and pump absorption distribution of LD corner-pumped laser with different parameters of Nd:YAG composite slab crystal. Resonator experiment was performed to suppress the oscillation of higher-order modes and realize TEM(00) operation. Experiment results showed that the output power was 11.94 W with an optical-optical efficiency of over 26%, and the M2 factors of beam quality at width and thickness directions were 1.18 and 1.34, respectively. It has been proven that the corner pumped configuration could obtain laser output with good beam quality as a result of high pump efficiency and good pump uniformity. PMID- 19997429 TI - Surface plasmon resonance and field enhancement in #-shaped gold wires metamaterial. AB - A #-shaped gold wires metamaterial is designed for surface enhanced Raman spectroscopy (SERS) and sensing. The tunability of surface plasmon resonance (SPR) excitations, hotspots distribution, localized field enhancement and sensitivity of the structure are investigated. In contrast to most metamaterial, the #-shaped structure exhibits two pronounced SPRs that are insensitive to the polarization of excitation light. Pure electromagnetic Raman enhancement factors of about 10(6) are achieved on the symmetrically distributed field hotspots. It is possible to break the usable wavelength range of conventional gold SERS substrates via higher order excitations of the #-shaped metamaterial. In addition, the sensitivity and the figure of merits are found to be comparable or even higher than those of conventional SERS substrates. All these factors together with the high reproducibility nature of metamaterial and its simple planer structure suggest that this structure is very promising for surface enhanced spectroscopy and sensing applications. PMID- 19997430 TI - Non-sequential optimization technique for a computer controlled optical surfacing process using multiple tool influence functions. AB - Optical surfaces can be accurately figured by computer controlled optical surfacing (CCOS) that uses well characterized sub-diameter polishing tools driven by numerically controlled (NC) machines. The motion of the polishing tool is optimized to vary the dwell time of the polisher on the workpiece according to the desired removal and the calibrated tool influence function (TIF). Operating CCOS with small and very well characterized TIF achieves excellent performance, but it takes a long time. This overall polishing time can be reduced by performing sequential polishing runs that start with large tools and finish with smaller tools. In this paper we present a variation of this technique that uses a set of different size TIFs, but the optimization is performed globally - i.e. simultaneously optimizing the dwell times and tool shapes for the entire set of polishing runs. So the actual polishing runs will be sequential, but the optimization is comprehensive. As the optimization is modified from the classical method to the comprehensive non-sequential algorithm, the performance improvement is significant. For representative polishing runs we show figuring efficiency improvement from approximately 88% to approximately 98% in terms of residual RMS (root-mean-square) surface error and from approximately 47% to approximately 89% in terms of residual RMS slope error. PMID- 19997431 TI - The general theory of phase shifting algorithms. AB - We have been reporting several new techniques of analysis and synthesis applied to Phase Shifting Interferometry (PSI). These works are based upon the Frequency Transfer Function (FTF) and how this new tool of analysis and synthesis in PSI may be applied to obtain very general results, among them; rotational invariant spectrum; complex PSI algorithms synthesis based on simpler first and second order quadrature filters; more accurate formulae for estimating the detuning error; output-power phase noise estimation. We have made our cases exposing these aspects of PSI separately. Now in the light of a better understanding provided by our past works we present and expand in a more coherent and holistic way the general theory of PSI algorithms. We are also providing herein new material not reported before. These new results are on; a well defined way to combine PSI algorithms and recursive linear PSI algorithms to obtain resonant quadrature filters. PMID- 19997432 TI - CO2 laser induced long period gratings in optical microfibers. AB - Long period gratings (LPGs) are fabricated by use of focused high frequency CO(2) laser pulses to periodically modify the transverse dimension of silica microfibers. A 20-period LPG with a 27 dB attenuation dip is realized in a microfiber with a diameter of approximately 6.3 microm. The resonant wavelength has a negative temperature coefficient and a high sensitivity to external refractive index. The microfiber LPGs may be useful in micron scale in-fiber devices and sensors. PMID- 19997433 TI - Devil's vortex-lenses. AB - In this paper we present a new kind of vortex lenses in which the radial phase distribution is characterized by the "devil's staircase" function. The focusing properties of these fractal DOEs coined Devil's vortex-lenses are analytically studied and the influence of the topological charge is investigated. It is shown that under monochromatic illumination a vortex devil's lens give rise a focal volume containing a delimited chain of vortices that are axially distributed according to the self-similarity of the lens. PMID- 19997434 TI - Resonance-enhanced optical forces between coupled photonic crystal slabs. AB - The behaviors of lateral and normal optical forces between coupled photonic crystal slabs are analyzed. We show that the optical force is periodic with displacement, resulting in stable and unstable equilibrium positions. Moreover, the forces are strongly enhanced by guided resonances of the coupled slabs. Such enhancement is particularly prominent near dark states of the system, and the enhancement effect is strongly dependent on the types of guided resonances involved. These structures lead to enhancement of light-induced pressure over larger areas, in a configuration that is directly accessible to externally incident, free-space optical beams. PMID- 19997435 TI - Complete compensation of pulse broadening in an amplifier-based slow light system using a nonlinear regeneration element. AB - We experimentally demonstrate complete compensation of pulse broadening in an amplifier-based slow light system. The configuration of the delay line basically consists of two stages: a conventional Brillouin slow light system and a nonlinear regeneration element. Signal pulses experienced both time delay and temporal broadening through the Brillouin delay line and then the delayed pulses were delivered into a nonlinear optical loop mirror. Due to the nonlinear response of the transmission of the fiber loop, the inevitably broadened pulses were moderately compressed in the output of the loop, without loss in the capacity to delay the pulses. The overall result is that, for the maximum delay, the width of the pulse could be kept below the input width while the time delays introduced by the slow light element were preserved. Using this delay line, a signal pulse with duration of 27 ns at full width at half maximum was delayed up to 1.3-bits without suffering from signal distortion. PMID- 19997436 TI - Trapping double negative particles in the ray optics regime using optical tweezers with focused beams. AB - The capabilities of optical tweezers to trap DNG (double negative) spherical particles, with both negative permittivity and permeability, are explored in detail by analyzing some interesting theoretical features not seeing in conventional DPS (double positive) particles possessing positive refractive index. The ray optics regime is adopted and, although this regime is quite simple and limited, its validity is already known and tested for DPS particles such as biological cells and molecules trapped by highly focused beams. Simulation results confirm that even for ray optics, DNG particles present unusual and interesting trapping characteristics. PMID- 19997437 TI - Adaptive improved element free Galerkin method for quasi- or multi-spectral bioluminescence tomography. AB - Bioluminescence tomography (BLT) has become a powerful tool for whole-body small animal imaging. In this contribution, an adaptive improved element free Galerkin method (IEFGM) is presented to perform a quantitative reconstruction of the internal light source using quasi- or multi-spectral information, which not only can avoid the time-consuming mesh generation but also can reduce the ill posedness of BLT effectively. In the algorithm, the reconstruction can be largely enhanced by an adaptive technology based on a posteriori error estimation. Finally, the numerical and physical phantom experiment results show that the bioluminescent source can be recovered accurately. PMID- 19997438 TI - A simple technique for accurate and complete characterisation of a Fabry-Perot cavity. AB - It has become a significant challenge to accurately characterise the properties of recently developed very high finesse optical resonators (F > 10(6)). A similar challenge is encountered when trying to measure the properties of cavities in which either the probing laser or the cavity length is intrinsically unstable. We demonstrate in this article the means by which the finesse, mode-matching, free spectral range, mirror transmissions and dispersion may be measured easily and accurately even when the laser or cavity has a relatively poor intrinsic frequency stability. PMID- 19997439 TI - Highly birefringent vaterite microspheres: production, characterization and applications for optical micromanipulation. AB - This paper reports on a simple synthesis and characterization of highly birefringent vaterite microspheres, which are composed of 20-30 nm sized nanocrystalls. Scanning electron microscopy shows a quite disordered assembly of nanocrystals within the microspheres. However, using optical tweezers, the effective birefringence of the microspheres was measured to be Deltan = 0.06, which compares to Deltan = 0.1 of vaterite single crystals. This suggests a very high orientation of the nanocrystals within the microspheres. A hyperbolic model of the direction of the optical axis throughout the vaterite spherulite best fits the experimental data. Results from polarized light microscopy further confirm the hyperbolic model. PMID- 19997440 TI - High repetition rate dual-rod acousto-optics Q-switched composite Nd:YVO4 laser. AB - We demonstrate the high-repetition rate acousto-optic Q-switching of a dual-rod Nd:YVO4 laser. Two thermally bonded composite YVO4-Nd:YVO4-YVO4 rod crystals were used as the gain medium to reduce the serious thermal effect. The pump mode and its influence on the laser mode were analyzed, and the effective pump radius was controlled by tuning the spectrum of laser diodes with adjustment on the temperature of laser diodes. Three different configurations of cavity optics were designed and the output performance using these cavities was investigated. In Q switching operation, 73.2 W TEM00 mode average power at 650 kHz was obtained. The stable Q-switching range was from 80 kHz to 650 kHz with the pulse duration increasing from 17.5 ns to 80 ns. In CW operation, 78 W TEM00 mode and 93 W multi mode output power was achieved with different cavity designs, corresponding to the optical-optical efficiency of 46.5% and 52.2% respectively. PMID- 19997441 TI - Simultaneous reference and differential waveform acquisition in time-resolved terahertz spectroscopy. AB - We present a new method for data acquisition in time-resolved terahertz spectroscopy experiments. Our approach is based on simultaneous collection of reference and differential THz scans. Both the optical THz generation beam and the pump beam are modulated at two different frequencies that are not harmonic with respect to each other. Our method allows not only twice as fast data acquisition but also minimization of noise connected to slowly varying laser power fluctuations and timing instabilities. Our use of the nonlinear crystal N benzyl-2-methyl-4-nitroaniline (BNA) enables time-resolved THz spectroscopy to beyond 5 THz, thereby highlighting that the presented method is especially valuable at higher frequencies where phase errors in the data acquisition become increasingly important. PMID- 19997442 TI - Thermo-optic locking of a semiconductor laser to a microcavity resonance. AB - We experimentally demonstrate thermo-optic locking of a semiconductor laser to an integrated toroidal optical microcavity. The lock is maintained for time periods exceeding twelve hours, without requiring any electronic control systems. Fast control is achieved by optical feedback induced by scattering centers within the microcavity, with thermal locking due to optical heating maintaining constructive interference between the cavity and the laser. Furthermore, the optical feedback acts to narrow the laser linewidth, with ultra high quality microtoroid resonances offering the potential for ultralow linewidth on-chip lasers. PMID- 19997443 TI - High-speed modulation of a compact silicon ring resonator based on a reverse biased pn diode. AB - High speed modulation based on a compact silicon ring resonator operating in depletion mode is demonstrated. The device exhibits an electrical small signal bandwidth of 19 GHz. The device is therefore a candidate for highly compact, wide bandwidth modulators for a variety of applications. PMID- 19997444 TI - Autonomous observations of in vivo fluorescence and particle backscatteringin an oceanic oxygen minimum zone. AB - The eastern South Pacific (ESP) oxygen minimum zone (OMZ) is a permanent hydrographic feature located directly off the coasts of northern Chile and Peru. The ESP OMZ reaches from coastal waters out to thousands of kilometers offshore, and can extend from the near surface to depths greater than 700 m. Oxygen minimum zones support unique microbial assemblages and play an important role in marine elemental cycles. We present results from two autonomous profiling floats that provide nine months of time-series data on temperature, salinity, dissolved oxygen, chlorophyll a, and particulate backscattering in the ESP OMZ. We observed consistently elevated backscattering signals within low-oxygen waters, which appear to be the result of enhanced microbial biomass in the OMZ intermediate waters. We also observed secondary chlorophyll a fluorescence maxima within low oxygen waters when the upper limit of the OMZ penetrated the base of the photic zone. We suggest that autonomous profiling floats are useful tools for monitoring physical dynamics of OMZs and the microbial response to perturbations in these areas. PMID- 19997445 TI - Wavelength tuning of planar photonic crystals by local processing of individual holes. AB - Tuning of the resonant wavelength of a single hole defect cavity in planar photonic crystals was demonstrated using transmission spectroscopy. Local post production processing of single holes in a planar photonic crystal is carried out after selectively opening a masking layer by focused ion beam milling. The resonance was blue-shifted by enlargement of selected holes using local wet chemical etching and red-shifted by infiltration with liquid crystals. This method can be applied to precisely control the resonant frequency, and can also be used for mode selective tuning. PMID- 19997446 TI - Anisotropy and particle-size effects in nanostructured plasmonic metamaterials. AB - We study the optical properties of metamaterials formed by layers of metallic nanoparticles. The effective optical constants of these materials are retrieved from the calculated angle-dependent Fresnel reflection coefficients for s and p incident-light polarization. We investigate the degree of anisotropy in the effective permittivity as a function of inter-layer spacing, particle size, filling fraction of the metal, and particle shape. For layers of spherical particles periodically arranged in a hexagonal lattice, the anisotropy disappears for the three inter-layer spacings corresponding to simple cubic (sc), bcc, and fcc volume symmetry. For non-spherical particles, an isotropic response can be still obtained with other values of the inter-layer spacing. Finally, we provide a quantitative answer to the question of how many layers are needed to form an effectively homogeneous metamaterial slab. Surprisingly, only one layer can be enough, except in the spectral range close to the particle plasmon resonances. PMID- 19997447 TI - Intense red-emitting Y4Al2O9:Eu3+ phosphor with short decay time and high color purity for advanced plasma display panel. AB - A new phosphor Y(4)Al(2)O(9):Eu(3+) (YAM:Eu(3+)) emitting intense monochromatic red at 612 nm under vacuum ultraviolet (VUV) and ultraviolet (UV) excitations has been developed for application in next generation plasma display panels (PDPs). The developed phosphor has better luminescence efficiency, colour purity and shorter decay time than commercial (Y,Gd)BO(3):Eu(3+) red emitting PDP phosphor. High color purity (x = 0.67, y = 0.32) under VUV excitation with short decay time (1.03 msec) and excellent stability against degradation during PDP panel preparation suggest that YAM:Eu(3+) is a potential candidate for present and future PDPs. Surface coating by SiO(2) further improved phosphor characteristics. PMID- 19997448 TI - THz photomixing synthesizer based on a fiber frequency comb. AB - A frequency doubled erbium doped modelocked fiber frequency comb is used to implement a THz photomixing synthesizer. The useful THz linewidth is in order of 150 kHz and has been assessed along with the frequency accuracy by spectroscopic measurements demonstrating a relative accuracy of 10(-8) at frequencies around 1 THz. The THz synthesizer is used to implement a THz spectrometer to study the rotational absorption spectrum of carbonyl sulfide (OCS). Measurement of the principal transitions between 813 GHz and 1283 GHz allowed the properties of the THz spectrometer to be compared with competing techniques, and demonstrates the potential of the THz photomixing synthesizer as an alternative means to explore the THz domain. PMID- 19997449 TI - Combined photoacoustic and ultrasound biomicroscopy. AB - We report on the development of an imaging system capable of combined ultrasound and photoacoustic imaging based on a fast-scanning single-element 25-MHz ultrasound transducer and a unique light-delivery system. The system is capable of 20 ultrasound frames per second and slower photoacoustic frame rates limited by laser pulse-repetition rates. Laser and ultrasound pulses are interlaced for co-registration of photoacoustic and ultrasound images. In vivo imaging of a human finger permits ultrasonic visualization of vessel structures and speckle changes indicative of blood flow, while overlaid photoacoustic images highlight some small vessels that are not clear from the ultrasound scan. Photoacoustic images provide optical absorption contrast co-registered in the structural and blood-flow context of ultrasound with high-spatial resolution and may prove important for clinical diagnostics and basic science of the microvasculature. PMID- 19997450 TI - Hartmann-Shack wavefront sensing for nonlinear materials characterization. AB - We present two new techniques exploiting a Hartmann-Shack wavefront sensor to characterize the optical self-focusing effect of nonlinear materials. We demonstrate that the defocus Zernike coefficient (C5) can be used to quantify nonlinear optical properties of materials. In the first technique proposed, the wavefront of a collimated laser beam transmitted through a nonlinear sample is analyzed with different irradiance values. In the second technique,instead of conventional detectors, a Hartmann- Shack sensor is used in a Z-scan setup. The methods are demonstrated by measuring the nonlinear refractive indices of CS2 and Quartz, using femtosecond Ti:sapphire lasers at 76 MHz and 1 KHz repetition rate. PMID- 19997451 TI - Detection of counterfeit U.S. paper money using intrinsic fluorescence lifetime. AB - Genuine U.S. Federal Reserve Notes have a consistent, two-component intrinsic fluorescence lifetime. This allows for detection of counterfeit paper money because of its significant differences in fluorescence lifetime when compared to genuine paper money. We used scanning two-photon laser excitation and the time correlated single photon counting (TCSPC) method to sample a approximately 4 mm(2) region. Three types of counterfeit samples were tested. Four out of the nine counterfeit samples fit to a one-component decay. Five out of nine counterfeit samples fit to a two-component model, but are identified as counterfeit due to significant deviations in the longer lifetime component compared to genuine bills. PMID- 19997452 TI - Conjugated polymer-fullerene blend with strong optical limiting in the near infrared. AB - Optical-quality, melt processable thick films of a conjugated polymer blend containing poly(2-methoxy-5-(2-ethyl-hexyloxy)-(phenylene vinylene)) (MEH-PPV), a C(60) derivative (PCBM) and a plasticizer (1,2-di-iso-octylphthalate) have been developed and their nonlinear absorption and optical limiting properties have been investigated. These blend materials exhibited strong optical limiting characteristics in the near infrared region (750-900 nm), with broad temporal dynamic range spanning femtosecond to nanosecond pulse widths. The dispersion of the optical limiting figure-of-merit of the MEH-PPV:PCBM:DOP blend shows a peak near the wavelength of the MEH-PPV cation, indicating an important role of one photon and two-photon induced charge transfer in the nonlinear absorption response. PMID- 19997453 TI - Blue light generated by intra-cavity frequency doubling of an edge-emitting diode laser with a periodically poled LiNbO3 crystal. AB - We demonstrate for the first time to our knowledge intra-cavity frequency doubling (ICFD) of an edge-emitter diode laser using a 10 mm-long 5.0 microm periodically poled LiNbO(3) (PPLN) crystal. An optical output power of 33 mW second harmonic blue light at 490.5 nm is generated at 1.0 A injection current, equivalent to an overall wall-plug efficiency of 1.8%. The measured M(2) values of blue beam are 1.7 and 2.4 along the fast and slow axis. PMID- 19997454 TI - A low bending loss multimode fiber transmission system. AB - This paper presents a high bend tolerant multimode optical fiber transmission system that is compatible with standard 50 microm graded index multimode fiber, in terms of achievable bandwidth and interconnectivity losses. When the 10 loops of the proposed bend resistive multimode fiber were wrapped around a cylinder of 1.5 mm radius, bend losses below -0.2 dB were achieved in case of experimentally produced fiber. Furthermore, when the section of the proposed bend resistive fiber was inserted between two sections of a standard 50 microm graded index multimode fiber, the total experimental measured loss proved to be below -0.15 dB. PMID- 19997455 TI - Region-restricted rapid keypoint registration. AB - A two-stage keypoint registration approach is proposed to achieve frame-rate performance, while maintaining high accuracy under large perspective and scale variations. First, an agglomerative clustering algorithm based on an effective edge significance measure is adopted to derive the corresponding regions for keypoint detection. Next, a light-weight detector and a compact descriptor are utilized to obtain the exact location of the keypoints. In conjunction with the point transferring method, the proposed approach can perform registration task in textureless regions robustly. Experiments are conducted to demonstrate that the approach can handle the real-time tracking tasks. PMID- 19997456 TI - Tri-material multilayer coatings with high reflectivity and wide bandwidth for 25 to 50 nm extreme ultraviolet light. AB - Magnesium/silicon carbide (Mg/SiC) multilayers have been fabricated with normal incidence reflectivity in the vicinity of 40% to 50% for wavelengths in the 25 to 50 nm wavelength range. However many applications, for example solar telescopes and ultrafast studies using high harmonic generation sources, desire larger bandwidths than provided by high reflectivity Mg/SiC multilayers. We investigate introducing a third material, Scandium, to create a tri-material Mg/Sc/SiC multilayer allowing an increase the bandwidth while maintaining high reflectivity. PMID- 19997457 TI - Lattice modes mediate radiative coupling in metamaterial arrays. AB - We show that a resonant response with very high quality factors can be achieved in periodic metamaterials by radiatively coupling their structural elements. The coupling is mediated by lattice modes and can be efficiently controlled by tuning the lattice periodicity. Using a recently developed terahertz (THz) near-field imaging technique and conventional far-field spectroscopy together with numerical simulations we pinpoint the underlying mechanisms. In the strong coupling regimes we identify avoided crossings between the plasmonic eigenmodes and the diffractive lattice modes. PMID- 19997458 TI - Microwave generation in an electro-absorption modulator integrated with a DFB laser subject to optical injection. AB - This paper presents a new technique to generate microwave signal using an electro absorption modulator (EAM) integrated with a distributed feedback (DFB) laser subject to optical injection. Experiments show that the frequency of the generated microwave can be tuned by changing the wavelength of the external laser or adjusting the bias voltage of the EAM. The frequency response of the EAM is studied and found to be unsmooth due to packaging parasitic effects and four-wave mixing effect occurring in the active layer of the DFB laser. It is also demonstrated that an EA modulator integrated in between two DFB lasers can be used instead of the EML under optical injection. This integrated chip can be used to realize a monolithically integrated tunable microwave source. PMID- 19997459 TI - Analytical study of optical bistability in silicon-waveguide resonators. AB - We present a theoretical model that describes accurately the nonlinear phenomenon of optical bistability in silicon-waveguide resonators but remains amenable to analytical results. Using this model, we derive a transcendental equation governing the intensity of a continuous wave transmitted through a Fabry-Perot resonator formed using a silicon-on-insulator waveguide. This equation reveals a dual role of free carriers in the formation of optical bistability in silicon. First, it shows that free-carrier absorption results in a saturation of the transmitted intensity. Second, the free-carrier dispersion and the thermo-optic effect may introduce phase shifts far exceeding those resulting from the Kerr effect alone, thus enabling one to achieve optical bistability in ultrashort resonators that are only a few micrometers long. Bistability can occur even when waveguide facets are not coated because natural reflectivity of the silicon- r interface can provide sufficient feedback. We find that it is possible to control the input-output characteristics of silicon-based resonators by changing the free carrier lifetime using a reverse-biased p-n junction. We show theoretically that such a technique is suitable for realization of electronically assisted optical switching at a fixed input power and it may lead to silicon-based, nanometer size, optical memories. PMID- 19997460 TI - A geometrical optics polarimetric bidirectional reflectance distribution function for dielectric and metallic surfaces. AB - A polarimetric bidirectional reflectance distribution function (pBRDF), based on geometrical optics, is presented. The pBRDF incorporates a visibility (shadowing/masking) function and a Lambertian (diffuse) component which distinguishes it from other geometrical optics pBRDFs in literature. It is shown that these additions keep the pBRDF bounded (and thus a more realistic physical model) as the angle of incidence or observation approaches grazing and better able to model the behavior of light scattered from rough, reflective surfaces. In this paper, the theoretical development of the pBRDF is shown and discussed. Simulation results of a rough, perfect reflecting surface obtained using an exact, electromagnetic solution and experimental Mueller matrix results of two, rough metallic samples are presented to validate the pBRDF. PMID- 19997461 TI - Incoherent "slow and fast light". AB - We show experimentally that the effects of pulse delay and advancement usually ascribed to the "slow and fast light" under conditions of the coherent population oscillations (CPO) can be universally observed with incoherent light fields on objects with the pure-intensity nonlinearity. As a light source, we used an incandescent lamp and as objects for study, a photochromic glass and a thermochromic coating. The response of the objects to intensity modulation of the incident light reproduced in all details the commonly accepted experimental evidences of the "light with a negative group velocity" and "ultraslow light". Thus we show that observations of the pulse delay (advancement) and characteristic changes in the light intensity modulation spectrum are not enough to make conclusion about modification of the light group velocity in the medium. PMID- 19997462 TI - Design of zero reference codes using cross-entropy method. AB - This paper considers the use of autocorrelation properties to design zero reference codes (ZRCs) for optical applications. Based on the properties of the autocorrelation function, the design of an optimum ZRC problem is transformed into a minimization problem with binary variables, and the objective is to minimize the second maximum of the autocorrelation signal sigma. However, the considerable computational complexity for an exhaustive search through all combinations of (nn (1)l) different code patterns is a potential problem especially for large codes, where n and n1 are the length of the ZRC and the number of transparent slits, respectively. To minimize sigma while reducing the computational complexity at the same time, we introduce the Cross-Entropy (CE) method, an effective algorithm that solves various combinatorial optimization problems to obtain a good code. The computer simulation results show that compared with the conventional genetic algorithm (GA), the proposed CE obtains the better sigma with low computational complexity. PMID- 19997463 TI - Strong exciton-photon coupling in inorganic-organic multiple quantum wells embedded low-Q microcavity. AB - Optoelectronic-compatible heterostructures are fabricated from layered inorganic organic multiple quantum wells (IO-MQW) of Cyclohexenyl ethyl ammonium lead iodide, (C(6)H(9)C(2)H(4)NH(3))(2)PbI(4) (CHPI). These hybrids possess strongly resonant optical features, are thermally stable and compatible with hybrid photonics assembly. Room-temperature strong-coupling is observed when these hybrids are straightforwardly embedded in metal-air (M-A) and metal-metal (M-M) low-Q microcavities, due to the large oscillator strength of these IO-MQWs. The strength of the Rabi splitting is 130 meV for M-A and 160 meV for M-M cavities. These values are significantly higher than for J-aggregates in all-metal microcavities of similar length. These experimental results are in good agreement with transfer matrix simulations based on resonant excitons. Incorporating exciton-switching hybrids allows active control of the strong-coupling parameters by temperature, suggesting new device applications. PMID- 19997464 TI - Geometry dependence of field enhancement in 2D metallic photonic crystals. AB - Geometry dependence of surface plasmon resonance of 2D metallic photonic crystals (PCs) was assessed using rigorous 3D finite difference time domain analysis. PCs of noble metallic rectangular and cylindrical nanopillars in square and triangular lattices on thick noble metal film were simulated for maximum field enhancement. It was found that the period, size and thickness of the nanopillars can be tuned to excite of surface plasmons at desired wavelengths in visible and near-infrared ranges. Maximum electric field enhancement near the nanopillars was found to be greater than 10X. The detail analysis of PCs tuned for 750 nm wavelength showed that thickness of nanopillars was the most sensitive parameter for field enhancement, and triangular lattice PCs had the wider enhancement bandwidth than square lattice PCs. Results showed that these PCs are sensitive with incident angle (theta) but not with polarization angle (phi). PMID- 19997466 TI - 170 Gbit/s transmission in an erbium-doped waveguide amplifier on silicon. AB - Signal transmission experiments were performed at 170 Gbit/s in an integrated Al(2)O(3):Er(3+) waveguide amplifier to investigate its potential application in high-speed photonic integrated circuits. Net internal gain of up to 11 dB was measured for a continuous-wave 1532 nm signal under 1480 nm pumping, with a threshold pump power of 4 mW. A differential group delay of 2 ps between the TE and TM fundamental modes of the 5.7-cm-long amplifier was measured. When selecting a single polarization open eye diagrams and bit error rates equal to those of the transmission system without the amplifier were observed for a 1550 nm signal encoded with a 170 Gbit/s return-to-zero pseudo-random 2(7)-1 bit sequence. PMID- 19997465 TI - Volumetric microvascular imaging of human retina using optical coherence tomography with a novel motion contrast technique. AB - Phase variance-based motion contrast imaging is demonstrated using a spectral domain optical coherence tomography system for the in vivo human retina. This contrast technique spatially identifies locations of motion within the retina primarily associated with vasculature. Histogram-based noise analysis of the motion contrast images was used to reduce the motion noise created by transverse eye motion. En face summation images created from the 3D motion contrast data are presented with segmentation of selected retinal layers to provide non-invasive vascular visualization comparable to currently used invasive angiographic imaging. This motion contrast technique has demonstrated the ability to visualize resolution-limited vasculature independent of vessel orientation and flow velocity. PMID- 19997467 TI - Pyroliton: pyroelectric spatial soliton. AB - The concept of optical beam self-trapping in pyroelectric photorefractive medium is presented. We show that the temperature controlled spontaneous polarisation of ferroelectric crystals produces an optical nonlinearity that can lead to formation of 2-D spatial soliton named pyroliton. Experimental demonstrations performed in lithium niobate crystals illustrate that efficient self-trapping occurs either for ordinary or extraordinary polarisation under moderate temperature increase. For instance, a 15 microm diameter pyroliton can be formed with a 10 degree temperature raise. PMID- 19997468 TI - Tunable delay slow-light in an active fiber Bragg grating. AB - We proposed and experimentally demonstrated an extremely simple and feasible slow light technique to achieve tunable optical delay by using the Er/Yb codoped fiber Bragg grating (FBG). The signal light experiences strong dispersion when it is launched into the reflection edge of FBG, and the group delay value is determined by the signal wavelength and the pump power. In the experiment, a controllable delay of 0.9 ns can be obtained through changing the 980 nm pump power. The group velocity can be slowed down to 5.6x10(7) m/s, which is 19% of the speed of light in free space. It provides a very simple approach to control the light group delay, which is likely to have important implications for practical applications. PMID- 19997469 TI - Optical and electrical properties of Au nanoparticles in two-dimensional networks:an effective cluster model. AB - We report a study of the optical and electric properties of Au nanoparticle networks grown on the porous alumina membrane by dry atom sputtering deposition approach. An effective cluster model was developed to evaluate the dielectric function and the electrical conductivities of the nanoparticle networks by taking into account the effects of the Au particle size, the Au volume fraction, and the particle-particle interaction. The calculated transmission spectra from the model were in good agreement with the experimental data. The percolation threshold of the as-fabricated structure was predicted to occur at Au volume fraction of 0.18, consistent with the dc electric resistance measurement. PMID- 19997471 TI - XPM and SBS nonlinear effects on MLSE with varying uncompensated dispersion. AB - We investigate the performance of a maximum likelihood sequence estimation (MLSE) receiver at 10.7 Gb/s in the presence of two optical nonlinear impairments, cross phase modulation (XPM) and stimulated Brillouin scattering (SBS). We find that the tolerance to both nonlinearities decreases with larger levels of uncompensated dispersion. Our results also suggest that the MSLE receiver loses its linear regime advantage in comparison to a standard receiver at some dispersion levels in the presence of the nonlinear effects. We demonstrate that long uncompensated links up to 160 km may show better tolerance to the nonlinear effects with a lower dispersion fiber when using an MLSE receiver. PMID- 19997470 TI - Magnetic field distribution of a highly focused radially-polarized light beam. AB - A simple expression for the magnetic filed of a highly focused radially polarized light is derived and the incorrect results for the time averaged Poynting vector and the trapping stability for a gold particle presented in the paper "Trapping metallic Rayleigh particles with radial polarization" by Zhan (Opt. Express 12, 3377-3382 (2004)) are corrected. PMID- 19997472 TI - A broadband ASE light source-based full-duplex FTTX/ROF transport system. AB - A full-duplex fiber-to-the-X (FTTX)/radio-over-fiber (ROF) transport system based on a broadband amplified spontaneous emission (ASE) light source is proposed and demonstrated for rural wide-spread villages. Combining the concepts of long transmission transmission and ring topology, a long-haul single-mode fiber (SMF) trunk is sharing with multiple rural villages. Externally modulated baseband (BB) (1.25 Gbps) and radio-frequency (RF) (622 Mbps/10 GHz) signals are successfully transmitted simultaneously. Good bit error rate (BER) performance was achieved to demonstrate the practice of providing wire/wireless connections for long-haul wide-spread rural villages. Since our proposed system uses only a broadband ASE light source to achieve multi-wavelengths transmissions, it also reveals an outstanding one with simpler and more economic advantages. PMID- 19997474 TI - Reduction of chromatic dispersion due to coupling for synchronized-router-based flat-passband filter using multiple-input arrayed waveguide grating. AB - An approach to reducing the chromatic dispersion due to coupling between input waveguides before the input slab for a synchronized-router-based flat-passband filter using a multiple-input arrayed waveguide grating (AWG) is proposed. The proposed method uses phase compensation at the waveguide array of the AWG by correction of waveguide lengths. The characteristics of the flat-passband filter that consists of a multiple-input AWG combined with cascaded Mach-Zehnder interferometers (MZIs) are simulated using a theoretical model of the multiple input AWG based on Fourier optics and the coupled-mode theory. The simulation result reveals that the chromatic dispersion within the passband can be significantly reduced by using phase compensation and additional dummy waveguides at the input just before the slab. PMID- 19997473 TI - A high spectral sensitivity interferometer based on the dispersive property of the semiconductor GaAs. AB - We develop an interferometer which has high spectral sensitivity based on the dispersive property of the semiconductor GaAs in the near-infrared region. Our experiment demonstrates that the spectral sensitivity could be greatly enhanced by adding a slow light medium into the interferometer and is proportional to the group index of the material. Subsequently the factors which influence the spectral sensitivity of the interferometer are analyzed. Moreover, we provide potential applications of such interferometers using the dispersive property of semiconductor in whole infrared region. PMID- 19997475 TI - Role of oocyte quality in meiotic maturation and embryonic development. AB - The quality of oocytes plays a key role in a proper embryo development. In humans, oocytes of poor quality may be the cause of women infertility and an important obstacle in successful in vitro fertilization (IVF). The competence of oocytes depends on numerous processes taking place during the whole oogenesis, but its final steps such as oocyte maturation, seem to be of key importance. In this paper, we overview factors involved in the development of a fully functional female gamete with Xenopus laevis as a major experimental model. Modern approaches, e.g. proteomic analysis, enable the identification of novel proteins involved in oocyte development. EP45, called also Seryp or pNiXa, which belongs to the serpin (serine protease inhibitors) super-family is one of such recently analyzed proteins. This protein seems to be involved in the stimulation of meiotic maturation and embryo development. EP45 is potentially a key factor in correct oocyte development and determining the quality of oocytes. PMID- 19997476 TI - The influence of mating on estrogen receptor alpha protein level in spleen and uterine macrophages in female mice. AB - Macrophages are antigen-presenting cells that have a key role in the regulation of immune phenomena and are responsible for the recognition of paternal antigens during pregnancy. The aim of this study was to investigate changes in the estrogen receptor alpha (ERalpha) protein level in splenic and uterine mature (F4/80(+)MHC II(+)) and immature (F4/80(+)MHC II(-) ) macrophages in female mice during time corresponding to the preimplantation period. C57BL/6J females in estrus were mated with Balb/c male mice or were mechanically stimulated through the vagina to achieve pseudopregnancy. Uterine and spleen cells were isolated on days 0.5 and 3.5 after mating or after uterine cervix stimulation. ERalpha content in macrophages was measured by flow cytometry and expressed as mean fluorescence intensity (MFI). The ERalpha level in splenic macrophages on 0.5 day after mating was higher than that in splenic macrophages of pseudopregnant mice on 0.5 day after stimulation. The ERalpha level was also higher in mature than in immature macrophages present in both the spleen and uterus, especially in mated mice. In the spleen, a correlation was found between the percentage of mature macrophages and ERalpha level in these cells. In conclusion, the elevated alpha level observed shortly after mating in splenic but not in uterine macrophages indicates an early systemic response to male antigens. PMID- 19997477 TI - Effect of naloxone treatment on luteinizing hormone and testosterone concentrations in boars with high and low libido. AB - The objective was to determine the effects of the opioid peptide receptor antagonist, naloxone on circulating concentrations of luteinizing hormone (LH) and testosterone in boars characterized as having high (n=8) or low libido (n=8) based on the willingness to mount an artificial sow and allow semen collection. On the day of the experiment, blood was sampled every 15 min for 4 h before and 4 h after i.v. injection of naloxone (1 mg/kg body weight). After naloxone treatment, a libido status by time interaction was detected and concentrations of LH within 15 min after treatment were greater (p<0.05) for High-libido boars than for Low-libido boars. Concentrations of testosterone were highly variable amongst boars and there were no effects of libido status (p=0.66) or libido status by time (p=0.66). There was, however, an effect of time (pA shift at position 998 of the ESR2 complete coding sequence in all tissue samples when compared to NCBI reference sequence U81451.1, but this polymorphism was not found in other ESR2 reference sequences. CONCLUSIONS: Our findings indicate that defects in the coding region of sex steroid receptors do not contribute to the pathogenesis of lacrimal gland disease in mouse models of Sjogren's syndrome. PMID- 19997527 TI - Aging and inflammation: etiological culprits of cancer. AB - The biochemical phenomenon of aging, as universal as it is, still remains poorly understood. A number of diseases are associated with aging either as a cause or consequence of the aging process. The incidence of human cancers increases exponentially with age and therefore cancer stands out as a disease that is intricately connected to the process of aging. Emerging evidence clearly suggests that there is a symbiotic relationship between aging, inflammation and chronic diseases such as cancer; however, it is not clear whether aging leads to the induction of inflammatory processes thereby resulting in the development and maintenance of chronic diseases or whether inflammation is the causative factor for inducing both aging and chronic diseases such as cancer. Moreover, the development of chronic diseases especially cancer could also lead to the induction of inflammatory processes and may cause premature aging, suggesting that longitudinal research strategies must be employed for dissecting the interrelationships between aging, inflammation and cancer. Here, we have described our current understanding on the importance of inflammation, activation of NF-kappaB and various cytokines and chemokines in the processes of aging and in the development of chronic diseases especially cancer. We have also reviewed the prevailing theories of aging and provided succinct evidence in support of novel theories such as those involving cancer stem cells, the molecular understanding of which would likely hold a great promise towards unraveling the complex relationships between aging, inflammation and cancer. PMID- 19997532 TI - Feasibility of Ecological Momentary Assessment Using Cellular Telephones in Methamphetamine Dependent Subjects. AB - BACKGROUND: Predictors of relapse to methamphetamine use are poorly understood. State variables may play an important role in relapse, but they have been difficult to measure at frequent intervals in outpatients. METHODS: We conducted a feasibility study of the use of cellular telephones to collect state variable data from outpatients. Six subjects in treatment for methamphetamine dependence were called three times per weekday for approximately seven weeks. Seven questionnaires were administered that assessed craving, stress, affect, and current type of location and social environment. RESULTS: 395/606 (65%) of calls attempted were completed. The mean time to complete each call was 4.9 (s.d. 1.8) minutes and the mean time to complete each item was 8.4 (s.d. 4.8) seconds. Subjects rated the acceptability of the procedures as good. All six cellular phones and battery chargers were returned undamaged. CONCLUSION: Cellular telephones are a feasible method for collecting state data from methamphetamine dependent outpatients. PMID- 19997531 TI - Building an evidence base on mental health interventions for children affected by armed conflict. AB - This paper reviews what is currently known from research about the effectiveness of interventions to address mental health problems in children and adolescents affected by armed conflict. The focus will be on interventions delivered in conflict affected countries either during active humanitarian emergencies or during the post conflict period. The paper will discuss two main paradigms of intervention dominating the field: psychosocial approaches and clinical/psychiatric approaches. The paper reviews some of the basic literature, theories and issues involved in assessment, programme planning, monitoring and evaluation of both approaches. In order to explore these issues in depth, the paper will draw from the author's field experiences with research in the Russian Federation and in northern Uganda. The paper also presents a brief review of a handful of other published evaluations of mental health interventions for war affected children. We will close with a discussion of what future research is needed to build an evidence base regarding mental health interventions for children affected by armed conflict as well as the ethical and feasibility issues associated with carrying out this work. PMID- 19997533 TI - Brief Therapy Based on Interrupting Ironic Processes: The Palo Alto Model. AB - The model of brief therapy developed by Fisch, Weakland, Watzlawick, and colleagues in Palo Alto is based on identifying and interrupting ironic processes that occur when repeated attempts to solve a problem keep the problem going or make it worse. Formulations of ironic problem-solution loops provide a template for assessment and strategic intervention, indicating where to look to understand what keeps a problem going (look for "more of the same" solution) and what needs to happen for the complaint to be resolved (someone must apply "less of the same" solution). Supporting research is preliminary but suggests this approach may be well suited for change-resistant clients. PMID- 19997535 TI - Characteristics of Activated Monocyte Phenotype Support R5-Tropic Human Immunodeficiency Virus. AB - BACKGROUND: Microbial translocation has been recognized as an important factor in monocyte activation and contributing to AIDS pathogenesis with elevated plasma lipopolysaccharide (LPS) levels, as a marker for microbial translocation, seen in advanced HIV disease. Therefore, the current study was undertaken to assess monocyte activation in vitro by LPS and to determine its impact on monocyte phenotype. METHODS: Monocytes from non-HIV-infected donors were analyzed for CD14, CD16, CD69, TNFalpha, and CCR5 by flow cytometry pre- and post-stimulation with LPS. In-vitro cultures were then set up to expose non-activated and activated monocytes to R5-, X4-, and dual (R5/X4)-tropic viruses; and the amount of HIV present on the cells was assayed. RESULTS: Non-HIV-infected monocytes, after LPS stimulation, were confirmed to have an activated phenotype with increase in CD16 and CD69 surface expressions (p<0.05). The activation phenotype was supported by increase in TNFalpha production, p<0.05. The activated monocytes had increased surface CCR5 (from 21% to 98%; p=0.05); and were found to have more R5-tropic virus than non-activated monocytes (p<0.05). CONCLUSIONS: Following activation by LPS, non-HIV-infected monocytes were found to have increase in surface CCR5. These activated monocytes, when exposed to R5-tropic virus, were found to have more virus compared to non-activated monocytes. The significance of the findings could lie in explaining how microbial translocation plays a role in HIV progression; and possibly promoting CCR5-directed strategies in treating HIV. PMID- 19997534 TI - Regenerative medicine strategies for treatment of neurogenic bladder. AB - Neurogenic bladder is a general term encompassing various neurologic dysfunctions in the bladder and external urethral sphincter caused by damage or disease. Therapeutic management options fall into the categories of conservative, minimally invasive or surgical. The current standard for surgical management is bladder augmentation using intestinal segments. However, because intestinal tissue possesses different functional characteristics to bladder tissue, numerous complications can ensue. Regenerative medicine uses combinations of cells and/or biomaterials to encourage regeneration of healthy tissue and offers an alternative approach for the replacement of lost or deficient organs, including the bladder. Promising results using the principles of regenerative medicine have already been obtained in children with neurogenic bladder caused by myelomeningocele. Human clinical trials, governed by the US FDA, are ongoing in the USA in both children and adults to further evaluate the safety and efficacy of this technology for regenerating bladders. More studies are in progress and additional advances in this field can be anticipated. PMID- 19997536 TI - Three Dimensional OCT in the Engineering of Tissue Constructs: A Potentially Powerful Tool for Assessing Optimal Scaffold Structure. AB - Optical Coherence Tomography (OCT) provides detailed, real-time information on the structure and composition of constructs used in tissue engineering. The focus of this work is the OCT three-dimensional assessment of scaffolding architecture and distribution of cells on it. PLGA scaffolds were imaged in two and three dimensions, both seeded and unseeded with cells. Then two types of scaffolds were reconstructed in three dimensions. Both scaffolding types were examined at three different seeding densities. The importance of three-dimensional assessments was evident, particularly with respect to porosity and identification of asymmetrical cell distribution. PMID- 19997537 TI - Engaging Client's Family and Friends in Online Counseling. AB - This paper describes how online counseling can be used to engage family members in the recovery of clients. The counselor guides the client and the family member separately through the stages of change, helping them commit to joint action. Each participant thinks through his/her concerns and the advantages of getting involved, each makes a specific commitment to working together under the guidance of the counselor. Finally each celebrates, with specific rituals, their decision to work together. Once the client and the family members commit to working together, the counselor guides them through the stages of change as a group. In the pre-contemplation stage, the family members are taught the goals of working together and the procedures to avoid dysfunctional conflict. In the contemplation phase, the client and the family members examine the shared environment and its links to target behavior. In the commitment to action stage, the client and the family members decide on specific changes in their shared environment. In the maintenance phase, the client and the family members analyze if the change has led to improvements. They analyze relapses and make additional cycles of improvement. PMID- 19997538 TI - SAPHO Syndrome Diagnosis and Treatment: Report of Five Cases and Review of the Literature. AB - BACKGROUND: The term "SAPHO (Synovitis, Acne, Pustulosis, Hyperostosis and Osteitis) syndrome" includes a variety of musculoskeletal disorders associated with skin conditions; Osteitis is the most prominent skeletal lesion, whereas palmoplantar pustulosis and acne are the main skin lesions. Diagnosing SAPHO syndrome is difficult, because this syndrome is often confused with suppurative osteomyelitis, which has similar clinical and pathologic findings. SAPHO diagnosis is even more difficult when atypical sites are involved and there are no skin lesions. PATIENTS AND METHODS: This case series presents five patients (3 women, 2 men), ages 27 to 44 years, who came to the Orthopaedic Department outpatient clinic for evaluation of pain in the humerus, clavicle, sacroiliac joints, and/or distal radius, and were diagnosed with SAPHO syndrome. Clinical and radiologic findings, treatment and outcome data, with up to 4 years of follow up are presented. An extensive discussion of the clinical presentation, published literature, treatment options and outcome of SAPHO syndrome is also included. RESULTS: Once the diagnosis of SAPHO syndrome was established, treatment with antibiotics (clindamycin) and non steroid anti-inflammatory drugs (lornoxicam) was remarkably effective. All patients did well and remained symptom free for up to four years, after a 3-8 month course of treatment. INTERPRETATION: SAPHO syndrome should be included in the differential diagnosis when evaluating patients with lytic, sclerotic, or hyperostotic bone lesions and pain. Prompt SAPHO syndrome recognition, followed by appropriate therapy with antibiotics and NSAIDs can produce rapid symptom resolution, while avoiding unnecessary procedures and longterm antibiotic therapy. PMID- 19997540 TI - Strategy for the treatment of clopidogrel low responsiveness in diabetes mellitus and stent implantation. PMID- 19997541 TI - Comparison of triple anti-platelet therapy (aspirin, clopidogrel, and cilostazol) and double anti-platelet therapy (aspirin and clopidogrel) on platelet aggregation in type 2 diabetic patients undergoing drug-eluting stent implantation. AB - BACKGROUND AND OBJECTIVES: Triple anti-platelet therapy may produce more potent inhibition of platelet aggregation in patients undergoing coronary stent implantation. We tested whether this effect could be maintained in diabetic patients, where platelet reactivity is increased and the risk of stent thrombosis is higher. SUBJECTS AND METHODS: Fifty five type 2 diabetic patients who had undergone drug-eluting stent (DES) implantation and chronic anti-platelet therapy (>1 month) were stratified according to the status of anti-platelet therapy. Platelet aggregation after adenosine diphosphate (ADP; 10 micromol/L and 20 micromol/L) stimulation was compared using light transmittance aggregometry between dual (aspirin plus clopidogrel, n=34) and triple therapy (aspirin, clopidogrel plus cilostazol, n=21) groups. RESULTS: The 2 groups had similar clinical and procedural characteristics. Maximal ADP-induced platelet aggregation was significantly lower in the triple therapy group than the dual therapy group (ADP 10 micromol/L, 37.1+/-15.4 vs. 28.3+/-11.8, p=0.03; ADP 20 micromol/L, 63.1+/-15.0 vs. 49.1+/-15.1, p=0.01), but there were no differences in diabetic treatment (oral hypoglycemic agent vs. insulin) or diabetic control {hemoglobin Alc (HbA1c) 7}. CONCLUSION: Triple anti-platelet therapy showed more potent inhibition of maximal ADP induced platelet aggregation in type 2 diabetic patients receiving chronic anti-platelet therapy. This finding suggests that triple antiplatelet therapy may be more effective in preventing thrombotic complications after DES implantation in type 2 diabetic patients. PMID- 19997542 TI - Long-term trend in the incidence of acute myocardial infarction in Korea: 1997 2007. AB - BACKGROUND AND OBJECTIVES: Information about disease incidence is indispensable for the active prevention and control of acute myocardial infarction (AMI). The purpose of this study was to provide basic information for the establishment of policy related to AMI by examining the long-term trends in incidence of AMI. SUBJECTS AND METHODS: This study identified the trend in disease incidence during between 1997 and 2007 using the Korean National Health Insurance Database that includes AMI {the 10th International Classification of Disease (ICD-10) code: I21, I22, I23, I250, I251} as a primary or secondary disease. RESULTS: The attack and incidence rates for AMI in 2007 were 118.4 and 91.8 per 100,000 persons, respectively, and the rates more than doubled for the 11 years. Both rates were higher among males than females and increased more in the older age groups. Incidence cases accounted for most of the total attack cases every year; however, in recent years the proportion of relapse cases was on the rise. The case fatality rate was highest (14.5%) in 2000, and declined rapidly to 9.8% in 2007. The case fatality rate was higher among females than males and the older age groups; in particular, female patients >/=65 years of age had the highest fatality rate. CONCLUSION: This study showed that AMI has been on the rise in Korea for 11 years. Therefore, the establishment of policy for intensive control of the incidence of AMI is necessary by building a continuous monitoring and surveillance system. PMID- 19997543 TI - Relationship between post-systolic motion during dobutamine stress echocardiography and functional recovery of myocardium after successful percutaneous coronary intervention. AB - BACKGROUND AND OBJECTIVES: Doppler myocardial imaging (DMI) has been suggested as a method of quantifying inducible ischemia during dobutamine stress echocardiography (DSE). Post-systolic motion (PSM) detected by DMI is related to peri-infarct ischemia during DSE. We hypothesized that PSM during DSE would predict recovery of dysfunctional myocardium after successful percutaneous coronary intervention (PCI). SUBJECTS AND METHODS: Thirty patients with dysfunctional myocardium in the left anterior descending coronary artery (LAD) territory were divided into two groups according to improvement of wall motion score index (WMSI) in the LAD territory at 6 months after successful PCI of the LAD. DMI was evaluated in the LAD territory during DSE. Fifteen patients showed improved WMSI (1.42+/-0.39) while the other 15 had unchanged WMSI (1.75+/-0.46) 1 month after PCI. Myocardial velocity was measured in the mid-septal, apico septal, and basal anterior segments of the LAD artery territory. PSM was defined as a positive wave appearing after the curve of systolic ejection had reached the zero line. RESULTS: Although there was no difference between resting PSMs in both groups, PSM during DSE was significantly higher in the improved WMSI group than in the WMSI group where it was unchanged. CONCLUSION: PSM during DSE predicts recovery of dysfunctional myocardium after successful PCI. PMID- 19997539 TI - Cardioembolic stroke in atrial fibrillation-rationale for preventive closure of the left atrial appendage. AB - Atrial fibrillation is the most common cardiac arrhythmias, and a major cause of morbidity and mortality due to cardioembolic stroke. The left atrial appendage is the major site of thrombus formation in non-valvular atrial fibrillation. Loss of atrial systole in atrial fibrillation and increased relative risk of associated stroke point strongly toward a role for stasis of blood in left atrial thrombosis, although thrombus formation is multifactorial, and much more than blood flow irregularities are implicated. Oral anticoagulation with vitamin-K antagonists is currently the most effective prophylaxis for stroke in atrial fibrillation. Unfortunately, this treatment is often contraindicated, particularly in the elderly, in whom risk of stroke is high. Moreover, given the risk of major bleeding, there is reason to be skeptical of the net benefit when warfarin is used in those patients. This work reviews the pathophysiology of cardioembolic stroke and critically spotlights the current status of preventive anticoagulation therapy. Various techniques to exclude the left atrial appendage from circulation were discussed as a considerable alternative for stroke prophylaxis. PMID- 19997544 TI - Ambulatory blood pressure monitoring and blood pressure load in obese children. AB - BACKGROUND AND OBJECTIVES: This study was aimed at evaluating the significance of blood pressure (BP) load in ambulatory blood pressure monitoring (ABPM) in obese children and adolescents. SUBJECTS AND METHODS: ABPM was conducted for 60 selected patients who had visited Sunlin Hospital between January 2008 and August 2008. Patients were classified into 3 groups; an obese group whose body mass index (BMI) was > the 95th percentile, an overweight group whose BMI was > the 85th percentile but less than the 95th percentile, and a normal group whose BMI was below the 85th percentile. Overall mean BP, day and night BP and BP load were measured by ABPM. RESULTS: Of the 60 patients, twenty-seven children belonged to the obese group, 9 and 24 to the overweight and the normal group, respectively. Among the three groups, the overall average systolic and diastolic BP, daytime diastolic BP, and systolic BP loads in daytime and nighttime were statistically different. Comparing the obese group with the normal group, systolic BP loads in daytime and nighttime in the obese group were significantly higher than those in the normal group. Also, the obese group had more patients whose BP loads were over 25% greater than the normal group while the difference in the number of patients with overall hypertension was not significantly different. CONCLUSION: Assessment of children's BP through assessment of BP load is a more detailed and precise tool than assessment through mean BP using ABPM and BP can be better controlled using measurement of BP load. PMID- 19997545 TI - The association between left ventricular hypertrophy and biomarkers in patients on continuous ambulatory peritoneal dialysis. AB - BACKGROUND AND OBJECTIVES: Left ventricular hypertrophy (LVH) is a major cardiovascular complication and an important predictor of mortality in patients with end stage renal disease. Some studies have shown that the serum aldosterone levels are correlated with LVH in non-diabetic patients undergoing hemodialysis. The objective of this study was to elucidate the relationships between serum biomarkers, including aldosterone, and echocardiographic findings, such as LVH, in patients on peritoneal dialysis. SUBJECTS AND METHODS: Thirty patients on continuous ambulatory peritoneal dialysis (CAPD) for >12 months at Soonchunhyang University Cheonan Hospital were included. Transthoracic echocardiography was performed and the left ventricular mass index (LVMI) was calculated using the Devereux formula. Serum biomarkers {N-terminal pro B-type natriuretic peptide (NT proBNP), troponin T, C-reactive protein, renin, and aldosterone} were measured. RESULTS: Sixteen of 30 patients had LVH on the basis of the LVMI. The mean serum aldosterone level was 62.53+/-60.73 pg/mL (range, 5.03-250.68 pg/mL). LVH, on the basis of the LVMI, was not correlated with the serum aldosterone level. The serum aldosterone levels were not associated with echocardiographic findings, even with co-existing diabetes mellitus. The LVMI had a negative correlation with the hemoglobin (r=-0.405, p=0.029) and hematocrit (r=-0.374, p=0.042), and a positive correlation with NT-proBNP (r=0.560, p=0.002). The other biomarkers (renin, aldosterone, troponin T, and C-reactive protein) were not correlated with the LVMI. The LVMI was correlated with the left atrium volume index (r=0.675, p<0.001). CONCLUSION: NT-proBNP is a good marker to predict LVH in patients undergoing CAPD. The serum aldosterone level is not correlated with LVMI, even with co-existing diabetes mellitus. PMID- 19997546 TI - Isolated left ventricular noncompaction cardiomyopathy accompanied by severe mitral regurgitation. AB - Isolated left ventricular noncompaction cardiomyopathy (IVNC) is a cardiomyopathy thought to be caused by arrest of normal embryogenesis of the endocardium and myocardium. This abnormality is often associated with other congenital cardiac defects. A 21-year-old man presented to the emergency department with worsening exertional dyspnea during the previous 2 months. Two-dimensional and Doppler echocardiography revealed an enlarged left atrium (LA) and a markedly dilated left ventricle (LV) with preserved LV systolic function, severe mitral valve regurgitation, and prolapse due to chordae rupture. The myocardium of the LV and right ventricle (RV) had excessively prominent trabeculations and deep intertrabecular recesses. He is the first patient in Korea who has undergone mitral valve replacement surgery because of severe mitral valve regurgitation and prolapse due to chordae rupture accompanied by IVNC. PMID- 19997547 TI - Effectiveness of the hugging balloon technique in coronary angioplasty for a heavy, encircling, calcified coronary lesion. AB - We report our experience in coronary angioplasty and intravascular ultrasonography (IVUS) on a heavy, encircling, calcified lesion that was not dilated with the use of a cutting balloon and a non-compliant balloon. The angioplasty was successfully performed with a simple and inexpensive hugging balloon technique. PMID- 19997548 TI - Kawasaki disease presenting as parotitis in a 3-month-old infant. AB - A male infant aged 3 months and 1 week had persistently high fever with parotitis that was unresponsive to antibiotics. Mumps was identified by serologic study, but he was finally diagnosed by clinical features as having Kawasaki disease and echocardiographic findings on the 9th day of fever. Parotitis, which is unresponsive to antibiotics, should be considered Kawasaki disease even though typical symptoms are not present. PMID- 19997549 TI - The preventive and therapeutic roles of regular physical activity. PMID- 19997552 TI - Complex and heterogeneous. PMID- 19997550 TI - Exercise as stroke prophylaxis. AB - BACKGROUND: Stroke is the third most common cause of death in industrialized countries, accounting for more than 10% of deaths over age 65. Most strokes are due to arteriosclerosis. Regular physical activity lowers arterial blood pressure and body weight and improves glucose and lipid metabolism, thereby slowing the development of arteriosclerosis and its cardiovascular complications, particularly myocardial infarction. This review focuses on the question whether physical activity might also have a preventive effect on cerebral infarction and hemorrhage. METHODS: This analysis is based on 33 prospective cohort studies and 10 case-control studies that addressed the potential effect of physical activity on stroke-related morbidity and mortality. RESULTS: Our meta-analysis shows that physical activity reduces the risk of all types of stroke (infarction, hemorrhage, and stroke of unspecified type). The relative risk (RR) of fatal or non-fatal cerebral infarction is 0.75, while the corresponding figures for cerebral hemorrhage and stroke of unspecified type are 0.67 and 0.71, respectively. The reduction of risk is only statistically significant for men. The case-control studies show an RR of 0.32 for men and women combined. CONCLUSIONS: When a multivariate analysis is performed that takes other vascular risk factors into account, physical activity is found to have an independent protective effect against cerebrovascular events. The effect is statistically significant only for men, not for women. PMID- 19997551 TI - Physical activity in the prevention and treatment of colorectal carcinoma. AB - BACKGROUND: Colorectal carcinoma is the most common type of tumor in Western countries. The risk of developing colorectal carcinoma depends both on genetic factors (familial predisposition) and on lifestyle-related factors such as body mass index, level of physical activity, and nutritional behavior. Regular physical activity is important in primary prevention, and there is also evidence that the prognosis after treatment of a colorectal carcinoma can be improved by exercise. METHODS: The PubMed database was searched for relevant articles that appeared in the last 10 years, and selected articles were evaluated. RESULTS: Cross-sectional studies have shown that regular physical activity (ca. 7 hours of brisk walking per week) lowers the risk of colon carcinoma by 40%. Physical activity also improves the outcome of patients already diagnosed with colorectal carcinoma: for example, patients with advanced disease (UICC stage II or III) have been found to survive significantly longer if they perform 4 hours of brisk walking per week, or the equivalent degree of physical exercise. CONCLUSIONS: Cross-sectional studies show that physically active persons are less likely to develop colorectal carcinoma than physically inactive persons, and that they have better outcomes in the event that they do develop the disease. The positive findings with respect to secondary prevention still need to be confirmed in interventional trials, but in primary prevention, at least, physical activity should be actively promoted, along with other beneficial lifestyle habits and screening measures. PMID- 19997553 TI - Definition should be changed. PMID- 19997554 TI - Meager result. PMID- 19997556 TI - Fertility desires and intentions of HIV-positive women of reproductive age in Ontario, Canada: a cross-sectional study. AB - BACKGROUND: Improvements in life expectancy and quality of life for HIV-positive women coupled with reduced vertical transmission will likely lead numerous HIV positive women to consider becoming pregnant. In order to clarify the demand, and aid with appropriate health services planning for this population, our study aims to assess the fertility desires and intentions of HIV-positive women of reproductive age living in Ontario, Canada. METHODOLOGY/PRINCIPAL FINDINGS: A cross-sectional study with recruitment stratified to match the geographic distribution of HIV-positive women of reproductive age (18-52) living in Ontario was carried out. Women were recruited from 38 sites between October 2007 and April 2009 and invited to complete a 189-item self-administered survey entitled "The HIV Pregnancy Planning Questionnaire" designed to assess fertility desires, intentions and actions. Logistic regression models were fit to calculate unadjusted and adjusted odds ratios of significant predictors of fertility intentions. The median age of the 490 participating HIV-positive women was 38 (IQR, 32-43) and 61%, 52%, 47% and 74% were born outside of Canada, living in Toronto, of African ethnicity and currently on antiretroviral therapy, respectively. Of total respondents, 69% (95% CI, 64%-73%) desired to give birth and 57% (95% CI, 53%-62%) intended to give birth in the future. In the multivariable model, the significant predictors of fertility intentions were: younger age (age<40) (p<0.0001), African ethnicity (p<0.0001), living in Toronto (p = 0.002), and a lower number of lifetime births (p = 0.02). CONCLUSIONS/SIGNIFICANCE: The proportions of HIV-positive women of reproductive age living in Ontario desiring and intending pregnancy were higher than reported in earlier North American studies. Proportions were more similar to those reported from African populations. Healthcare providers and policy makers need to consider increasing services and support for pregnancy planning for HIV-positive women. This may be particularly significant in jurisdictions with high levels of African immigration. PMID- 19997557 TI - Scrapie affects the maturation cycle and immune complex trapping by follicular dendritic cells in mice. AB - Transmissible spongiform encephalopathies (TSEs) or prion diseases are infectious neurological disorders of man and animals, characterised by abnormal disease associated prion protein (PrP(d)) accumulations in the brain and lymphoreticular system (LRS). Prior to neuroinvasion, TSE agents often accumulate to high levels within the LRS, apparently without affecting immune function. However, our analysis of scrapie-affected sheep shows that PrP(d) accumulations within the LRS are associated with morphological changes to follicular dendritic cells (FDCs) and tingible body macrophages (TBMs). Here we examined FDCs and TBMs in the mesenteric lymph nodes (MLNs) of scrapie-affected mice by light and electron microscopy. In MLNs from uninfected mice, FDCs could be morphologically categorised into immature, mature and regressing forms. However, in scrapie affected MLNs this maturation cycle was adversely affected. FDCs characteristically trap and retain immune complexes on their surfaces, which they display to B-lymphocytes. In scrapie-affected MLNs, some FDCs were found where areas of normal and abnormal immune complex retention occurred side by side. The latter co-localised with PrP(d) plasmalemmal accumulations. Our data suggest this previously unrecognised morphology represents the initial stage of an abnormal FDC maturation cycle. Alterations to the FDCs included PrP(d) accumulation, abnormal cell membrane ubiquitin and excess immunoglobulin accumulation. Regressing FDCs, in contrast, appeared to lose their membrane-attached PrP(d). Together, these data suggest that TSE infection adversely affects the maturation and regression cycle of FDCs, and that PrP(d) accumulation is causally linked to the abnormal pathology observed. We therefore support the hypothesis that TSEs cause an abnormality in immune function. PMID- 19997558 TI - Expression-based network biology identifies alteration in key regulatory pathways of type 2 diabetes and associated risk/complications. AB - Type 2 diabetes mellitus (T2D) is a multifactorial and genetically heterogeneous disease which leads to impaired glucose homeostasis and insulin resistance. The advanced form of disease causes acute cardiovascular, renal, neurological and microvascular complications. Thus there is a constant need to discover new and efficient treatment against the disease by seeking to uncover various novel alternate signalling mechanisms that can lead to diabetes and its associated complications. The present study allows detection of molecular targets by unravelling their role in altered biological pathways during diabetes and its associated risk factors and complications. We have used an integrated functional networks concept by merging co-expression network and interaction network to detect the transcriptionally altered pathways and regulations involved in the disease. Our analysis reports four novel significant networks which could lead to the development of diabetes and other associated dysfunctions. (a) The first network illustrates the up regulation of TGFBRII facilitating oxidative stress and causing the expression of early transcription genes via MAPK pathway leading to cardiovascular and kidney related complications. (b) The second network demonstrates novel interactions between GAPDH and inflammatory and proliferation candidate genes i.e., SUMO4 and EGFR indicating a new link between obesity and diabetes. (c) The third network portrays unique interactions PTPN1 with EGFR and CAV1 which could lead to an impaired vascular function in diabetic nephropathy condition. (d) Lastly, from our fourth network we have inferred that the interaction of beta-catenin with CDH5 and TGFBR1 through Smad molecules could contribute to endothelial dysfunction. A probability of emergence of kidney complication might be suggested in T2D condition. An experimental investigation on this aspect may further provide more decisive observation in drug target identification and better understanding of the pathophysiology of T2D and its complications. PMID- 19997559 TI - P-glycoprotein acts as an immunomodulator during neuroinflammation. AB - BACKGROUND: Multiple sclerosis is an inflammatory demyelinating disease of the central nervous system in which autoreactive myelin-specific T cells cause extensive tissue damage, resulting in neurological deficits. In the disease process, T cells are primed in the periphery by antigen presenting dendritic cells (DCs). DCs are considered to be crucial regulators of specific immune responses and molecules or proteins that regulate DC function are therefore under extensive investigation. We here investigated the potential immunomodulatory capacity of the ATP binding cassette transporter P-glycoprotein (P-gp). P-gp generally drives cellular efflux of a variety of compounds and is thought to be involved in excretion of inflammatory agents from immune cells, like DCs. So far, the immunomodulatory role of these ABC transporters is unknown. METHODS AND FINDINGS: Here we demonstrate that P-gp acts as a key modulator of adaptive immunity during an in vivo model for neuroinflammation. The function of the DC is severely impaired in P-gp knockout mice (Mdr1a/1b-/-), since both DC maturation and T cell stimulatory capacity is significantly decreased. Consequently, Mdr1a/1b -/- mice develop decreased clinical signs of experimental autoimmune encephalomyelitis (EAE), an animal model for multiple sclerosis. Reduced clinical signs coincided with impaired T cell responses and T cell-specific brain inflammation. We here describe the underlying molecular mechanism and demonstrate that P-gp is crucial for the secretion of pro-inflammatory cytokines such as TNF alpha and IFN-gamma. Importantly, the defect in DC function can be restored by exogenous addition of these cytokines. CONCLUSIONS: Our data demonstrate that P gp downmodulates DC function through the regulation of pro-inflammatory cytokine secretion, resulting in an impaired immune response. Taken together, our work highlights a new physiological role for P-gp as an immunomodulatory molecule and reveals a possible new target for immunotherapy. PMID- 19997560 TI - Combined bezafibrate and medroxyprogesterone acetate: potential novel therapy for acute myeloid leukaemia. AB - BACKGROUND: The majority of acute myeloid leukaemia (AML) patients are over sixty years of age. With current treatment regimens, survival rates amongst these, and also those younger patients who relapse, remain dismal and novel therapies are urgently required. In particular, therapies that have anti-leukaemic activity but that, unlike conventional chemotherapy, do not impair normal haemopoiesis. PRINCIPAL FINDINGS: Here we demonstrate the potent anti-leukaemic activity of the combination of the lipid-regulating drug bezafibrate (BEZ) and the sex hormone medroxyprogesterone acetate (MPA) against AML cell lines and primary AML cells. The combined activity of BEZ and MPA (B/M) converged upon the increased synthesis and reduced metabolism of prostaglandin D(2) (PGD(2)) resulting in elevated levels of the downstream highly bioactive, anti-neoplastic prostaglandin 15-deoxy Delta(12,14) PGJ(2) (15d-PGJ(2)). BEZ increased PGD(2) synthesis via the generation of reactive oxygen species (ROS) and activation of the lipid peroxidation pathway. MPA directed prostaglandin synthesis towards 15d-PGJ(2) by inhibiting the PGD(2) 11beta -ketoreductase activity of the aldo-keto reductase AKR1C3, which metabolises PGD(2) to 9alpha11beta-PGF(2alpha). B/M treatment resulted in growth arrest, apoptosis and cell differentiation in both AML cell lines and primary AML cells and these actions were recapitulated by treatment with 15d-PGJ(2). Importantly, the actions of B/M had little effect on the survival of normal adult myeloid progenitors. SIGNIFICANCE: Collectively our data demonstrate that B/M treatment of AML cells elevated ROS and delivered the anti neoplastic actions of 15d-PGJ(2). These observations provide the mechanistic rationale for the redeployment of B/M in elderly and relapsed AML. PMID- 19997561 TI - Identification of MAMDC1 as a candidate susceptibility gene for systemic lupus erythematosus (SLE). AB - BACKGROUND: Systemic lupus erythematosus (SLE) is a complex autoimmune disorder with multiple susceptibility genes. We have previously reported suggestive linkage to the chromosomal region 14q21-q23 in Finnish SLE families. PRINCIPAL FINDINGS: Genetic fine mapping of this region in the same family material, together with a large collection of parent affected trios from UK and two independent case-control cohorts from Finland and Sweden, indicated that a novel uncharacterized gene, MAMDC1 (MAM domain containing glycosylphosphatidylinositol anchor 2, also known as MDGA2, MIM 611128), represents a putative susceptibility gene for SLE. In a combined analysis of the whole dataset, significant evidence of association was detected for the MAMDC1 intronic single nucleotide polymorphisms (SNP) rs961616 (P -value = 0.001, Odds Ratio (OR) = 1.292, 95% CI 1.103-1.513) and rs2297926 (P -value = 0.003, OR = 1.349, 95% CI 1.109-1.640). By Northern blot, real-time PCR (qRT-PCR) and immunohistochemical (IHC) analyses, we show that MAMDC1 is expressed in several tissues and cell types, and that the corresponding mRNA is up-regulated by the pro-inflammatory cytokines tumour necrosis factor alpha (TNF-alpha) and interferon gamma (IFN-gamma) in THP-1 monocytes. Based on its homology to known proteins with similar structure, MAMDC1 appears to be a novel member of the adhesion molecules of the immunoglobulin superfamily (IgCAM), which is involved in cell adhesion, migration, and recruitment to inflammatory sites. Remarkably, some IgCAMs have been shown to interact with ITGAM, the product of another SLE susceptibility gene recently discovered in two independent genome wide association (GWA) scans. SIGNIFICANCE: Further studies focused on MAMDC1 and other molecules involved in these pathways might thus provide new insight into the pathogenesis of SLE. PMID- 19997563 TI - Spontaneous corneal perforation in a patient with lamellar ichthyosis and dry eye. AB - We report spontaneous corneal perforation in a patient with lamellar ichthyosis. The patient presented with complaints of pain, redness, diminished vision, and discharge in her right eye for 15 days. Visual acuities were light perception in the right and 20/400 in the left eye. Cicatricial ectropion in both lower eyelids and 2 mm perforation site in the center of the right cornea were observed. Lamellar ichthyosis was suspected because of scaling and excessive dryness of entire body skin and was confirmed by skin biopsy. Amniotic membrane transplantation and transient tarsorraphy was performed and systemic anti ichthyosis therapy was started. The follow-up visits were not possible because of patient inconsistency. In patients with cicatricial ectropion secondary to ichthyosis, corneal health should be closely monitored because of the perforation risk. PMID- 19997564 TI - Eye muscle surgery for infantile nystagmus syndrome in the first two years of life. AB - PURPOSE: To report visual and elctrophysioloigcal effects of eye muscle surgery in young patients with infantile nystagmus syndrome (INS). METHODS: Prospective, interventional case cohort of 19 patients aged under 24 months who were operated on for combinations of strabismus, an anomalous head posture, and nystagmus. All patients were followed at least nine months. Outcome measures, part of an institutionally approved study, included Teller acuity, head position, strabismic deviation, and eye movement recordings, from which waveform types and a nystagmus optimal foveation fraction (NOFF). Computerized parametric and nonparametric statistical analysis of data were perfomed using standard software on both individual and group data. RESULTS: Age averaged 17.7 months (13.1-month follow up). Thirteen (68%) patients had associated optic nerve or retinal disease. 42% had amblyopia, 68% had refractive errors. Group means in binocular Teller acuity (P < 0.05), strabismic deviation (P < 0.05), head posture (P < 0.001), and the NOFF measures (P < 0.01) from eye movement recordings improved in all patients. There was a change in null zone waveforms to more favorable jerk types. There were no reoperations or surgical complications. CONCLUSIONS: Surgery on the extraocular muscles in patients aged less than two years with INS results in improvements in multiple aspects of ocular motor and visual function. PMID- 19997562 TI - Original encounter with antigen determines antigen-presenting cell imprinting of the quality of the immune response in mice. AB - BACKGROUND: Obtaining a certain multi-functionality of cellular immunity for the control of infectious diseases is a burning question in immunology and in vaccine design. Early events, including antigen shuttling to secondary lymphoid organs and recruitment of innate immune cells for adaptive immune response, determine host responsiveness to antigens. However, the sequence of these events and their impact on the quality of the immune response remain to be elucidated. Here, we chose to study Modified Vaccinia virus Ankara (MVA) which is now replacing live Smallpox vaccines and is proposed as an attenuated vector for vaccination strategies against infectious diseases. METHODOLOGY/PRINCIPAL FINDINGS: We analyzed in vivo mechanisms triggered following intradermal (i.d.) and intramuscular (i.m.) Modified Vaccinia virus Ankara (MVA) administration. We demonstrated significant differences in the antigen shuttling to lymphoid organs by macrophages (MPhis), myeloid dendritic cells (DCs), and neutrophils (PMNs). MVA i.d. administration resulted in better antigen distribution and more sustained antigen-presenting cells (APCs) recruitment into draining lymph nodes than with i.m. administration. These APCs, which comprise both DCs and MPhis, were differentially involved in T cell priming and shaped remarkably the quality of cytokine-producing virus-specific T cells according to the entry route of MVA. CONCLUSIONS/SIGNIFICANCE: This study improves our understanding of the mechanisms of antigen delivery and their consequences on the quality of immune responses and provides new insights for vaccine development. PMID- 19997565 TI - Functional microperimetry and SD-OCT confirm consecutive retinal atrophy from optic nerve pit. AB - A congenital anomaly, optic nerve pit is often associated with serous retinal detachment involving macula. Long standing serous detachment leads to outer retinal atrophy and decrease in visual sensitivity. Recently, spectral-domain optical coherence tomography (OCT) has been reported to demonstrate a communication between the optic nerve sheath and the subretinal space. Vitreous cavity is proposed as an alternate source of fluid for accumulation in the subretinal space. We imaged a patient with optic nerve pit with Spectralis OCT and report the findings seen including the presence of an area of peripapapillary retinal atrophy, due to the spontaneous resolution of associated long-standing retinal detachment. PMID- 19997566 TI - Efficacy and safety of travoprost/timolol vs dorzolamide/timolol in patients with open-angle glaucoma or ocular hypertension. AB - PURPOSE: To compare the intraocular pressure- (IOP-) lowering efficacy of fixed combinations travoprost 0.004%/timolol 0.5% and dorzolamide 2%/timolol 0.5% in patients with ocular hypertension or open-angle glaucoma. METHODS: In this prospective, multicenter, double-masked, randomized clinical trial, 319 qualifying patients received either travoprost/timolol once daily in the morning (n = 157) or dorzolamide/timolol twice daily (n = 162). IOP was assessed morning and evening at 2 and 6 weeks. The primary outcome measure was mean diurnal IOP. RESULTS: Baseline mean IOP values were similar between groups. Mean pooled diurnal IOP was significantly lower in the travoprost/timolol group (16.5 mmHg +/ 0.23) than in the dorzolamide/timolol group (17.3 mmHg +/- 0.23; P = 0.011). Mean IOP was significantly lower in the travoprost/timolol group compared to the dorzolamide/timolol group at the 9 AM time point both at Week 2 (P = 0.006) and Week 6 (P = 0.002). The travoprost/timolol combination produced mean IOP reductions from baseline of 35.3% to 38.5%, while the dorzolamide/timolol combination produced mean IOP reductions from baseline of 32.5% to 34.5%. CONCLUSIONS: The fixed combination travoprost 0.004%/timolol 0.5% dosed once daily in the morning demonstrated superior mean diurnal IOP-lowering efficacy compared to dorzolamide 2%/timolol 0.5% dosed twice daily in patients with ocular hypertension or open-angle glaucoma. PMID- 19997567 TI - First-year treatment costs among new initiators of topical prostaglandin analogs. AB - OBJECTIVE: To estimate first-year costs among new initiators of topical prostaglandin analogs in a managed care population. RESEARCH DESIGN AND METHODS: We developed a model to estimate first-year direct medical costs. We derived treatment patterns from a claims database analysis. Published studies were used to estimate visit-related resource use. Costs were obtained from standard sources. RESULTS: The database analysis identified 9,063 patients meeting study criteria, 41% (n = 3,672) of whom remained on their initial prostaglandin therapy for 12 months after initiation. Adjunctive intraocular pressure lowering therapy was needed in 20.7%, 16.5%, 13.9%, and 8.9% of bimatoprost, latanoprost, travoprost, and BAK-free travoprost patients, respectively. Median numbers of days to the first prescription filled for adjunctive therapy (if required) were 69.5, 67.0, 123.0, and 158.5 for patients initiating on bimatoprost, latanoprost, travoprost, and BAK-free travoprost. Total estimated first-year costs were $1,457, $1,360, $1,278, and $1,307 for patients initiating therapy with bimatoprost, latanoprost, travoprost, and BAK-free travoprost. Findings were consistent through sensitivity analysis. CONCLUSIONS: A BAK-free prostaglandin analog may permit longer duration of monotherapy and be associated with lower first-year direct medical costs. Use of a claims database and the selection of new initiators of prostaglandin analogs limit projecting findings to all glaucoma patients. PMID- 19997568 TI - Characteristics of respondents with glaucoma and dry eye in a national panel survey. AB - BACKGROUND: There is an increasing body of evidence strongly suggesting that glaucoma medications may contribute to ocular surface disease and development of dry eye. OBJECTIVE: To identify glaucoma patients with dry eye, using a nationally representative sample, and to compare clinical and treatment characteristics with controls without dry eye. METHODS: Patients taking intraocular pressure-lowering medications were identified from the Medical Expenditure Panel Survey. A matched cohort without glaucoma served as controls. Dry eye was identified by diagnosis or use of prescription or over-the-counter medications. Demographic and clinical characteristics and medication use patterns were compared. RESULTS: The analysis identified 629 respondents with glaucoma and 6,934 controls without glaucoma. Dry eye was more common among glaucoma respondents than nonglaucoma controls (16.5% vs 5.6%, P < 0.0001). There was a nonsignificant trend for respondents with dry eye to report higher rates of glaucoma adjunctive therapy use compared to those without dry eye (44.2% vs 35.0%, P < 0.076). Prostaglandin analogs were the most common glaucoma medication. CONCLUSIONS: This analysis found that the rate of dry eye was higher in patients with glaucoma than in controls. The use of glaucoma adjunctive therapies may increase the rate of dry eye in glaucoma patients. PMID- 19997569 TI - Seasonal hematocrit variation and health risks in the adult population of Kinshasa, Democratic Republic of Congo. AB - Hematocrit (Hct) as an indicator of blood viscosity and mean arterial blood pressure (MAP) were assessed according to the season in adult participants of health screenings conducted throughout Kinshasa, Democratic Republic of Congo. Data was collected at the end of summer (April) and the end of winter (August) and identified by gender. Male Hcts in August were significantly higher (P < 0.0001) than in April (48.3% +/- 4.2% and 45.7% +/- 2.3%, respectively) while male MAP (85.0 +/- 8.4 mm Hg) was identical to that recorded in April (85.4 +/- 7.7 mm Hg). August female Hcts (41.4% +/- 3.1%) were statistically higher than those recorded in April (39.6% +/- 1.9%, P = 0.001), MAP being 82.3 +/- 7.3 vs 87.9 +/- 6.6 mm Hg, respectively (P = 0.0001). Systolic and diastolic blood pressures, heart rate, body mass indices, ages, and personal and familial medical histories of the August and April groups were not significantly different. This study offers further support for the assertion that the relationship between blood viscosity and pressure of a healthy population shows that increased Hct, and therefore increased blood viscosity is associated with lowered MAP, and presumably peripheral vascular resistance. PMID- 19997570 TI - Low use of statins and other coronary secondary prevention therapies in primary and secondary care in India. AB - OBJECTIVE: To determine the frequency of use of pharmacotherapy with aspirin, beta blocker, statin, and angiotensin-converting enzyme (ACE) inhibitor in patients with stable coronary heart disease (CHD) among physicians at different levels of health care in Rajasthan state, India. METHODS: Physicians practicing at tertiary hospitals and clinics at tertiary, secondary and primary levels were contacted. Prescriptions of CHD patients were audited and descriptive statistics reported. RESULTS: We evaluated 2,993 prescriptions (tertiary hospital discharge 711, tertiary 688, secondary 1,306, and primary 288). Use of aspirin was in 2,713 (91%) of prescriptions, beta blockers 2,057 (69%), ACE inhibitors or angiotensin receptor blockers (ARBs) 2,471 (82%), and statins 2,059 (69%). Any one of these drugs was prescribed in 2,991 (100%), any two in 2,880 (96%), any three in 1,740 (58%), and all four in 1,062 (35.5%) (P < 0.001). As compared to tertiary hospital, prescriptions at tertiary, secondary, and primary levels were lower: aspirin (96% vs 95%, 91%, 67%), beta blockers (80% vs 62%, 66%, 70%), statins (87% vs 82%, 62%, 21%): two drugs (98% vs 96%, 98%, 85%), three drugs (75% vs 58%, 55%, 28%), or four drugs (54% vs 44%, 28%, 7%) (P < 0.01). Use of ACE inhibitors/ARBs was similar while nitrates (43% vs 23%, 43%, 70%), dihydropyridine calcium channel blockers (12% vs 15%, 30%, 47%), and multivitamins (6% vs 26%, 37%, 47%) use was more in secondary and primary care. CONCLUSIONS: There is suboptimal use of various evidence-based drugs (aspirin, beta blockers, ACE inhibitors, and statins) for secondary prevention of CHD in India. PMID- 19997571 TI - Mitigating micro-and macro-vascular complications of diabetes beginning in adolescence. AB - Diabetes is a chronic disorder, which manifests when insulin levels or resistance to insulin action becomes insufficient to control systemic glucose levels. Although the number of available agents to manage diabetes continues to expand rapidly, the maintenance of euglycemia by individuals with diabetes remains a substantial challenge. Unfortunately, many patients with type 1 and type 2 diabetes will ultimately experience diabetes complications. These complications result from the toxic effects of chronic hyperglycemia combined with other metabolic derangements that afflict persons with diabetes. This review will present a comprehensive look at the complications of diabetes, the risk factors for their progression, the mechanistic basis for their development, and the clinical approach to screening for, preventing, and treating these sequelae. In addition, since diabetes is commonly diagnosed in childhood, we will provide a special focus on the care of the adolescent patient. PMID- 19997572 TI - Endothelial repair capacity and apoptosis are inversely related in obstructive sleep apnea. AB - PURPOSE: To investigate the impact of obstructive sleep apnea (OSA) on endothelial repair capacity and apoptosis in the absence of potentially confounding factors including obesity. PATIENTS AND METHODS: Sixteen patients with a body mass index <30 and newly diagnosed OSA and 16 controls were studied. Circulating levels of endothelial progenitor cells, a marker of endothelial repair capacity, and endothelial microparticles, a marker of endothelial apoptosis, were quantified before and after four-week therapy with continuous positive airway pressure (CPAP). Endothelial cell apoptotic rate was also quantified in freshly harvested venous endothelial cells. Vascular reactivity was measured by flow-mediated dilation. RESULTS: Before treatment, endothelial microparticle levels were greater and endothelial progenitor cell levels were lower in patients with OSA than in controls (P < 0.001 for both). Levels of endothelial microparticles and progenitors cells were inversely related (r = 0.67, P < 0.001). Endothelial progenitor cell levels increased after effective treatment (P = 0.036). CONCLUSIONS: In the absence of any co-morbid conditions including obesity, OSA alone impairs endothelial repair capacity and promotes endothelial apoptosis. These early endothelial alterations may underlie accelerated atherosclerosis and increased cardiovascular risk in OSA. PMID- 19997573 TI - Critical appraisal of the role of pitavastatin in treating dyslipidemias and achieving lipid goals. AB - Pitavastatin is a potent HMG-CoA reductase inhibitor and efficient hepatocyte low density lipoprotein cholesterol (LDL-C) receptor inducer, producing robust reduction of the serum LDL-C levels, even at a low dose. Pitavastatin and its lactone form are minimally metabolized by CYP enzymes, and are therefore associated with minimal drug-drug interactions (DDIs). Pitavastatin 2 to 4 mg has potent LDL-C-reducing activity, equivalent to that of atorvastatin 10 to 20 mg; several clinical trials have revealed consistently superior high-density lipoprotein cholesterol (HDL-C) elevating activity of pitavastatin than that of atorvastatin. Pitavastatin-induced HDL-C elevation has been shown to be sustained, even incremental, in long-term clinical trials. Pitavastatin was as well-tolerated as atorvastatin or simvastatin in double-blind randomized clinical trials. Two-year long-term safety and effectiveness of pitavastain has been confirmed in a large-scale, prospective post-marketing surveillance. The safety and efficacy profile of pitavastatin is favorable for the treatment of dyslipidemia, especially in metabolic syndrome patients. In addition to control of LDL-C, adequate control of triglyceride (TG) and HDL-C, hypertension and hyperglycemia is also necessary in metabolic syndrome patients. Pitavastatin produces adequate control of LDL-C and TG, along with potent and incremental HDL C elevation, with a low frequency of DDIs. PMID- 19997575 TI - Comparative assessment of angiotensin receptor blockers in different clinical settings. AB - Cardiovascular and renal disease can be regarded as progressing along a sort of continuum which starts with cardiovascular risk factors (hypertension, diabetes, dyslipidemia, smoking, etc), evolves with progression of atherosclerotic lesions and organ damage, and then becomes clinically manifest with the major clinical syndromes (myocardial infarction, stroke, heart failure, end-stage renal disease). The blood pressure control remains a fundamental mechanism for prevention of cardiovascular disease. The renin-angiotensin system is believed to play an important role along different steps of the cardiovascular disease continuum. Convincing evidence accumulated over the last decade that therapeutic intervention with angiotensin receptor blockers (ARBs) is effective to slow down or block the progression of cardiovascular disease at different steps of the continuum, with measurable clinical benefits. However, despite the shared mechanism of action, each ARB is characterized by specific pharmacological properties that may influence its clinical efficacy. Indeed, important differences among available ARBs emerged from clinical studies. Therefore, generalization of results obtained with a specific ARB to all available ARBs may be misleading. The present review provides a comparative assessment of the different ARBs in their efficacy on major clinical endpoints along the different steps of the cardiovascular disease continuum. PMID- 19997576 TI - The treatment of diabetic foot infections: focus on ertapenem. AB - Clinically, 3 distinct stages of diabetic foot infection may be recognized: localized infection, spreading infection and severe infection. Each of these presentations may be complicated by osteomyelitis. Infection can be caused by Gram-positive aerobic, and Gram-negative aerobic and anaerobic bacteria, singly or in combination. The underlying principles are to diagnose infection, culture the bacteria responsible and treat aggressively with antibiotic therapy. Localized infections with limited cellulitis can generally be treated with oral antibiotics on an outpatient basis. Spreading infection should be treated with systemic antibiotics. Severe deep infections need urgent admission to hospital for wide-spectrum intravenous antibiotics. Clinical and microbiological response rates have been similar in trials of various antibiotics and no single agent or combination has emerged as most effective. Recently, clinical and microbiological outcomes for patients treated with ertapenem were equivalent to those for patients treated with piperacillin/tazobactam. It is also important to judge the need for debridement and surgery, to assess the arterial supply to the foot and consider revascularization either by angioplasty or bypass if the foot is ischemic. It is also important to achieve metabolic control. Thus infection in the diabetic foot needs full multidisciplinary treatment. PMID- 19997577 TI - Aspirin and clopidogrel hyporesponsiveness and nonresponsiveness in patients with coronary artery stenting. AB - Patients undergoing coronary artery stenting receive an antiplatelet regimen to reduce the risk of antithrombotic complications. Current guidelines recommend the use of acetyl salicylic acid (aspirin) and clopidogrel as evidenced by large clinical trials. There has been a concern about variable responses of patients to aspirin and clopidogrel which may predispose them to subacute stent thrombosis or late stent thrombosis. Up to 25% of patients with acute myocardial infarction (AMI) undergoing percutaneous coronary intervention (PCI) were found to have hyporesponsiveness or resistance to clopidogrel which may predispose them to recurrent events. Dual antiplatelet regimen is a standard therapy in these patients and there is always a concern about variable responses to aspirin and clopidogrel predisposing them to acute coronary syndrome (ACS). Prevalence of this hyporesponsiveness or resistance may be due to noncompliance, genetic mutations, co-morbid situations and concomitant use of other drugs. This issue is of considerable importance in the era of coronary drug eluting stents when a long term dual antiplatelet regimen is needed. This paper is a review for clinicians taking care of such patients with hyporesponsiveness or nonresponsiveness to dual antiplatelet regimen. PMID- 19997579 TI - Significance of initial blood pressure and comorbidity for the efficacy of a fixed combination of an angiotensin receptor blocker and hydrochlorothiazide in clinical practice. AB - BACKGROUND: Two-thirds of all patients with arterial hypertension need drug combinations to achieve blood pressure (BP) goals. Fixed combinations have high efficacy and result in high patient compliance. 300 mg irbesartan plus 25 mg hydrochlorothiazide (HCTZ) has been investigated only in clinical trials but not in daily practice. METHODS: A multicenter, noninterventional, noncontrolled observational study with 8123 patients seen by 1604 physicians in daily practice. BP reduction (office measurements), co-morbid disease and tolerability were documented over a 6-month observational period. RESULTS: At mean baseline BP of 161 +/- 15/94 +/- 10 mmHg, administering of fixed combination resulted in a substantial BP reduction averaging 28 +/- 15/14 +/- 10 mmHg (P < 0.001). Decrease of systolic BP ran parallel with increasing systolic baseline BP (Spearman's Rho 0.731; P < 0.0001; diastolic BP vs diastolic baseline BP Rho 0.740; P < 0.0001), independent from patient characteristics (age, obesity, diabetes or nephropathy) but enhanced with short history of hypertension (P < 0.0001 vs long history), prior beta blockers (P = 0.001 vs prior angiotensin receptor blockers [ARBs]), prior calcium channel blockers (P = 0.046 vs prior ARBs) and no prior medication (P = 0.012 vs prior ARBs). High compliance (>98%) and low incidence of adverse events (0.66%) were documented. CONCLUSIONS: The fixed combination of 300 mg irbesartan with 25 mg HCTZ was efficacious and tolerable in an unselected patient population in primary care. PMID- 19997580 TI - NADPH oxidase expression and production of superoxide by human corneal stromal cells. AB - PURPOSE: Superoxide (O(2) (.-)) may function as a second messenger or regulator of signal transduction when produced at low concentrations in the proper locations within cells. The purpose of these studies was to determine whether human corneal stromal (HCS) fibroblasts are capable of producing O(2) (.-) via nicotinamide adenine dinucleotide phosphate (NADPH) oxidases, a family of protein complexes believed to be responsible for the localized and limited production of O(2) (.-) with regulatory activity. METHODS: HCS cells, grown as primary and low passage cultures of fibroblasts, were used as the sources of RNA for reverse transcriptase PCR, with primers specific for mRNAs encoding the proteins that comprise NADPH oxidases. Small interfering (si)RNAs were used to knockdown specific NOX mRNAs. Proteins composing the NADPH oxidase complexes were identified using western blots. The production of O(2) (.-) by whole cells and cell-free preparations was assessed by measurement of NADPH-dependent superoxide dismutase-inhibitable cytochrome c reduction. RESULTS: Whole cells and cell-free extracts of corneal stromal fibroblasts produced O(2) (.-) in an NADPH-dependent manner. These fibroblasts constitutively produced mRNAs encoding eight proteins known to comprise NADPH oxidase complexes. mRNAs encoding NOX1, NOX4, NOX5, p22 phox, p47 phox, p67 phox, and p40 phox as well as Rac were expressed. Treatment of HCS fibroblasts with siRNA pools specific for each of these three NOXs significantly reduced the steady state levels of the respective mRNAs. Western blots confirmed the existence of all the proteins required for O(2) (.-) production. Rac 1, a regulator of the activity of some forms of NADPH complexes was present in membranous cell fractions containing the oxidase proteins. CONCLUSIONS: HCS fibroblasts produced O(2) (.-) in a NADPH-dependent manner via at least three isoforms of NADPH oxidase. These cells expressed NOX1, NOX4, NOX5, p22 phox, p47 phox, p67 phox, and p40 phox as well as Rac. SiRNAs directed against each of the three putative isoforms of NOX significantly reduced the steady state levels of the appropriate NOX mRNA pools, thus confirming the existence of the three isoforms. The O(2) (.-) produced by the NADPH oxidases in HCS fibroblasts is a potential contributor to signal transduction pathways and a regulator of gene expression as well as a potential participant in processes that occur during inflammation. PMID- 19997582 TI - Strategies for nanoplasmonic core-satellite biomolecular sensors: Theory-based Design. AB - We present a systematic theoretical study of core-satellite gold nanoparticle assemblies using the Generalized Multiparticle Mie formalism. We consider the importance of satellite number, satellite radius, the core radius, and the satellite distance, and we present approaches to optimize spectral shift due to satellite attachment or release. This provides clear strategies for improving the sensitivity and signal-to-noise ratio for molecular detection, enabling simple colorimetric assays. We quantify the performance of these strategies by introducing a figure of merit. In addition, we provide an improved understanding of the nanoplasmonic interactions that govern the optical response of core satellite nanoassemblies. PMID- 19997581 TI - Phenotypic characterisation and ZEB1 mutational analysis in posterior polymorphous corneal dystrophy in a New Zealand population. AB - PURPOSE: Posterior Polymorphous Dystrophy (PPCD) is a genetically heterogeneous corneal dystrophy, with linkage to three different chromosomal loci, with several genes in these loci being implicated. The role of both VSX1 and COL8A2 in PPCD remains controversial but recent work suggests that mutations in the transcription factor gene ZEB1/TCF8 account for disease in up to 30% of subjects, with a significant association with connective tissue abnormalities. This study aimed to determine the phenotype and contribution of ZEB1 mutations in a New Zealand PPCD population METHODS: Following informed consent, 11 probands with PPCD underwent extensive clinical characterization; including a questionnaire to determine birth history, general health, and the incidence of connective tissue abnormalities, slit lamp examination, photography and in vivo confocal microscopy. Family members were recruited where available. Biological specimens underwent mutational analysis of all nine coding exons of ZEB1. RESULTS: ZEB1 mutational analysis identified one mutation in the 11 probands (9.1%), a novel mutation in the initiating methionine of exon 1, c.1A-->G that results in the protein change p.Met1Val, with resultant aberrant initiation of translation. This mutation segregated with disease in the family, and was not present in 100 control chromosomes. No other ZEB1 mutations were observed in this cohort. CONCLUSION: Recent studies suggest that ZEB1 mutations may account for PPCD in 18 to 30% of cases, with the majority of the mutations in exons 5 and 7. Clinical and molecular analyses in this New Zealand cohort show a much lower incidence of ZEB1 sequence change, confirming the genetic heterogeneity of PPCD. We also report identification of a novel mutation in the initiating methionine that removes the Kozak sequence, thereby altering the site of initiation translation. PMID- 19997578 TI - Vascular health in children and adolescents: effects of obesity and diabetes. AB - The foundations for cardiovascular disease in adults are laid in childhood and accelerated by the presence of comorbid conditions, such as obesity, diabetes, hypertension, and dyslipidemia. Early detection of vascular dysfunction is an important clinical objective to identify those at risk for subsequent cardiovascular morbidity and events, and to initiate behavioral and medical interventions to reduce risk. Typically, cardiovascular screening is recommended for young adults, especially in people with a family history of cardiovascular conditions. Children and adolescents were once considered to be at low risk, but with the growing health concerns related to sedentary lifestyle, poor diet and obesity, cardiovascular screening may be needed earlier so that interventions to improve cardiovascular health can be initiated. This review describes comorbid conditions that increase cardiovascular risk in youth, namely obesity and diabetes, and describes noninvasive methods to objectively detect vascular disease and quantify vascular function and structure through measurements of endothelial function, arterial compliance, and intima-media thickness. Additionally, current strategies directed toward prevention of vascular disease in these populations, including exercise, dietary interventions and pharmacological therapy are described. PMID- 19997583 TI - Soluble state high resolution atomic force microscopy study of Alzheimer's beta amyloid oligomers. AB - We report here the direct observation of high resolution structures of assemblies of Alzheimer beta-amyloid oligomers and monomers using liquid atomic force microscopy (AFM). Visualization of nanoscale features of Abeta oligomers (also known as ADDLs) was carried out in tapping mode AFM in F12 solution. Our results indicate that ADDL preparations exist in solution primarily as a mixture of monomeric peptides and higher molecular mass oligomers. Our study clearly reveals that the size and shape of these oligomer aggregates exhibit a pronounced dependence on concentration. These studies show that wet AFM enables direct assessment of oligomers in physiological fluids and suggests that this method may be developed to visualize Abeta oligomers from human fluids. PMID- 19997584 TI - Effect of pressure on electrospray characteristics. AB - An experimental study of pulsating electrosprays operated at subambient pressure is reported. The pressure domain that affords stable electrospray operation appears to be limited by the vapor pressure of the liquid. The voltage driving the electrospray is shown to have a logarithmic dependence on pressure. The observed scaling amends the relationship currently used to calculate the electric field at the tip of the meniscus of an electrified liquid. PMID- 19997585 TI - On ghost authorship and reviews: the 6th International Congress on Peer Review and Biomedical Publication. PMID- 19997586 TI - The diagnosis and treatment of hypertensive disorders of pregnancy: new findings for antenatal and inpatient care. AB - BACKGROUND: Hypertensive disorders of pregnancy (HDP) are among the leading causes of maternal and fetal morbidity and mortality. New guidelines and findings from clinical trials must be taken into account so that the diagnosis and treatment of HDP can be optimized. METHODS: Current guidelines, Cochrane reviews, metaanalyses, and randomized, controlled trials were retrieved by a search in PubMed and the Cochrane Library for reports published from 2006 to March 2009. These publications were then analyzed and evaluated for their evidence levels (EL). RESULTS AND CONCLUSIONS: Aside from hypertension and proteinuria, the definition of preeclampsia (PE) should also take organ dysfunction into account. Important aspects of antenatal care include the following: the early recognition of risk factors, measurement of the uterine arteries in the 1st and 2nd trimesters with Doppler ultrasonography (A diagnostic tool which is now well established), prophylactic oral administration of 100 mg of acetylsalicylic acid daily from the beginning of pregnancy, particularly in high-risk patients (EL I++), and appropriate measurement of blood pressure and urinary protein. Patients should be hospitalized whenever indicated. Therapeutic goals are adequate treatment of hypertension, as well as seizure prophylaxis with magnesium sulphate in severe preeclampsia to prevent maternal cerebrovascular complications (EL I++). If delivery is indicated, it should be performed, regardless of the gestational age (EL IV). Careful monitoring during the puerperium and a general medical review six weeks after delivery are essential. Women with preeclampsia have a significantly elevated long-term risk of developing cardiovascular diseases in later life (EL I++). PMID- 19997587 TI - Principles of pediatric emergency care. AB - BACKGROUND: Children account for only a small percentage of pre-hospital emergency patients but are a special challenge for the treating physician. METHODS: The Medline database was selectively searched for articles appearing up to June 2009. The authors added other important literature of which they were aware. RESULTS: The broad spectrum of diseases, the wide age range with the physiological and anatomical changes that occur in it, and the special psychological, emotional, and communicative features of children make pediatric emergencies a special challenge for emergency physicians. CONCLUSIONS: A mastery of basic emergency techniques including clinical evaluation of the child, establishment of venous access, airway management, resuscitation, and drug dosing is essential for the successful emergency treatment of children. We recommend classifying the common non-traumatic pediatric emergencies by four cardinal manifestations: respiratory distress, altered consciousness, seizure, and shock. Classifying these rare emergency situations in this way helps assure that their treatment will be goal-oriented and appropriate to the special needs of sick children. PMID- 19997589 TI - Let's go down to brass tacks. PMID- 19997590 TI - Comparative sequence analysis of the non-protein-coding mitochondrial DNA of inbred rat strains. AB - The proper function of mammalian mitochondria necessitates a coordinated expression of both nuclear and mitochondrial genes, most likely due to the co evolution of nuclear and mitochondrial genomes. The non-protein coding regions of mitochondrial DNA (mtDNA) including the D-loop, tRNA and rRNA genes form a major component of this regulated expression unit. Here we present comparative analyses of the non-protein-coding regions from 27 Rattus norvegicus mtDNA sequences. There were two variable positions in 12S rRNA, 20 in 16S rRNA, eight within the tRNA genes and 13 in the D-loop. Only one of the three neutrality tests used demonstrated statistically significant evidence for selection in 16S rRNA and tRNA-Cys. Based on our analyses of conserved sequences, we propose that some of the variable nucleotide positions identified in 16S rRNA and tRNA-Cys, and the D loop might be important for mitochondrial function and its regulation. PMID- 19997591 TI - Combined anti-angiogenic therapy targeting PDGF and VEGF receptors lowers the interstitial fluid pressure in a murine experimental carcinoma. AB - Elevation of the interstitial fluid pressure (IFP) of carcinoma is an obstacle in treatment of tumors by chemotherapy and correlates with poor drug uptake. Previous studies have shown that treatment with inhibitors of platelet-derived growth factor (PDGF) or vascular endothelial growth factor (VEGF) signaling lowers the IFP of tumors and improve chemotherapy. In this study, we investigated whether the combination of PDGFR and VEGFR inhibitors could further reduce the IFP of KAT-4 human carcinoma tumors. The tumor IFP was measured using the wick-in needle technique. The combination of STI571 and PTK/ZK gave an additive effect on the lowering of the IFP of KAT-4 tumors, but the timing of the treatment was crucial. The lowering of IFP following combination therapy was accompanied by vascular remodeling and decreased vascular leakiness. The effects of the inhibitors on the therapeutic efficiency of Taxol were investigated. Whereas the anti-PDGF and anti-VEGF treatment did not significantly inhibit tumor growth, the inhibitors enhanced the effect of chemotherapy. Despite having an additive effect in decreasing tumor IFP, the combination therapy did not further enhance the effect of chemotherapy. Simultaneous targeting of VEGFR and PDGFR kinase activity may be a useful strategy to decrease tumor IFP, but the timing of the inhibitors should be carefully determined. PMID- 19997592 TI - Accurate and reliable cancer classification based on probabilistic inference of pathway activity. AB - With the advent of high-throughput technologies for measuring genome-wide expression profiles, a large number of methods have been proposed for discovering diagnostic markers that can accurately discriminate between different classes of a disease. However, factors such as the small sample size of typical clinical data, the inherent noise in high-throughput measurements, and the heterogeneity across different samples, often make it difficult to find reliable gene markers. To overcome this problem, several studies have proposed the use of pathway-based markers, instead of individual gene markers, for building the classifier. Given a set of known pathways, these methods estimate the activity level of each pathway by summarizing the expression values of its member genes, and use the pathway activities for classification. It has been shown that pathway-based classifiers typically yield more reliable results compared to traditional gene-based classifiers. In this paper, we propose a new classification method based on probabilistic inference of pathway activities. For a given sample, we compute the log-likelihood ratio between different disease phenotypes based on the expression level of each gene. The activity of a given pathway is then inferred by combining the log-likelihood ratios of the constituent genes. We apply the proposed method to the classification of breast cancer metastasis, and show that it achieves higher accuracy and identifies more reproducible pathway markers compared to several existing pathway activity inference methods. PMID- 19997593 TI - Integrative discovery of epigenetically derepressed cancer testis antigens in NSCLC. AB - BACKGROUND: Cancer/testis antigens (CTAs) were first discovered as immunogenic targets normally expressed in germline cells, but differentially expressed in a variety of human cancers. In this study, we used an integrative epigenetic screening approach to identify coordinately expressed genes in human non-small cell lung cancer (NSCLC) whose transcription is driven by promoter demethylation. METHODOLOGY/PRINCIPAL FINDINGS: Our screening approach found 290 significant genes from the over 47,000 transcripts incorporated in the Affymetrix Human Genome U133 Plus 2.0 expression array. Of the top 55 candidates, 10 showed both differential overexpression and promoter region hypomethylation in NSCLC. Surprisingly, 6 of the 10 genes discovered by this approach were CTAs. Using a separate cohort of primary tumor and normal tissue, we validated NSCLC promoter hypomethylation and increased expression by quantitative RT-PCR for all 10 genes. We noted significant, coordinated coexpression of multiple target genes, as well as coordinated promoter demethylation, in a large set of individual tumors that was associated with the SCC subtype of NSCLC. In addition, we identified 2 novel target genes that exhibited growth-promoting effects in multiple cell lines. CONCLUSIONS/SIGNIFICANCE: Coordinated promoter demethylation in NSCLC is associated with aberrant expression of CTAs and potential, novel candidate protooncogenes that can be identified using integrative discovery techniques. These findings have significant implications for discovery of novel CTAs and CT antigen directed immunotherapy. PMID- 19997594 TI - Therapeutic and adverse effects of a non-steroidal glucocorticoid receptor ligand in a mouse model of multiple sclerosis. AB - BACKGROUND: Dissociating glucocorticoid receptor (GR) ligands hold great promise for treating inflammatory disorders since it is assumed that they exert beneficial activities mediated by transrepression but avoid adverse effects of GR action requiring transactivation. Here we challenged this paradigm by investigating 2-(4-acetoxyphenyl)-2-chloro-N-methyl-ethylammonium chloride (CpdA), a dissociating non-steroidal GR ligand, in the context of experimental autoimmune encephalomyelitis (EAE), an animal model of multiple sclerosis (MS). METHODOLOGY/PRINCIPAL FINDINGS: CpdA inhibited pro-inflammatory mediators in myelin-specific T cells and fibroblasts in a GR-dependent manner while gene activation was abolished. However, it also induced massive apoptosis in various cell types even in the absence of the GR by engaging a Bcl-2- and caspase dependent pathway. (1)H NMR spectroscopy corroborated these findings by revealing that CpdA dissolved in buffered solutions rapidly decomposes into aziridine intermediates known to act as alkylating pro-apoptotic agents. Importantly, the dichotomy of CpdA action also became evident in vivo. Administration of high-dose CpdA to mice was lethal while treatment of EAE with low to intermediate amounts of CpdA dissolved in water significantly ameliorated the disease. The beneficial effect of CpdA required expression of the GR in T cells and was achieved by down regulating LFA-1 and CD44 on peripheral Th cells and by repressing IL-17 production. CONCLUSIONS/SIGNIFICANCE: CpdA has significant therapeutic potential although adverse effects severely compromise its application in vivo. Hence, non steroidal GR ligands require careful analysis prior to their translation into new therapeutic concepts. PMID- 19997595 TI - Orientation of mitotic spindles during the 8- to 16-cell stage transition in mouse embryos. AB - BACKGROUND: Asymmetric cell divisions are involved in the divergence of the first two lineages of the pre-implantation mouse embryo. They first take place after cell polarization (during compaction) at the 8-cell stage. It is thought that, in contrast to many species, spindle orientation is random, although there is no direct evidence for this. METHODOLOGY/PRINCIPAL FINDINGS: Tubulin-GFP and live imaging with a spinning disk confocal microscope were used to directly study spindle orientation in whole embryos undergoing the 8- to 16-cell stage transition. This approach allowed us to determine that there is no predetermined cleavage pattern in 8-cell compacted mouse embryos and that mitotic spindle orientation in live embryo is only modulated by the extent of cell rounding up during mitosis. CONCLUSIONS: These results clearly demonstrate that spindle orientation is not controlled at the 8- to 16-cell transition, but influenced by cell bulging during mitosis, thus reinforcing the idea that pre-implantation development is highly regulative and not pre-patterned. PMID- 19997596 TI - Restoration of full-length SMN promoted by adenoviral vectors expressing RNA antisense oligonucleotides embedded in U7 snRNAs. AB - BACKGROUND: Spinal Muscular Atrophy (SMA) is an autosomal recessive disease that leads to specific loss of motor neurons. It is caused by deletions or mutations of the survival of motor neuron 1 gene (SMN1). The remaining copy of the gene, SMN2, generates only low levels of the SMN protein due to a mutation in SMN2 exon 7 that leads to exon skipping. METHODOLOGY/PRINCIPAL FINDINGS: To correct SMN2 splicing, we use Adenovirus type 5-derived vectors to express SMN2-antisense U7 snRNA oligonucleotides targeting the SMN intron 7/exon 8 junction. Infection of SMA type I-derived patient fibroblasts with these vectors resulted in increased levels of exon 7 inclusion, upregulating the expression of SMN to similar levels as in non-SMA control cells. CONCLUSIONS/SIGNIFICANCE: These results show that Adenovirus type 5-derived vectors delivering U7 antisense oligonucleotides can efficiently restore full-length SMN protein and suggest that the viral vector mediated oligonucleotide application may be a suitable therapeutic approach to counteract SMA. PMID- 19997597 TI - A proteomic view of an important human pathogen--towards the quantification of the entire Staphylococcus aureus proteome. AB - The genome sequence is the "blue-print of life," but proteomics provides the link to the actual physiology of living cells. Because of their low complexity bacteria are excellent model systems to identify the entire protein assembly of a living organism. Here we show that the majority of proteins expressed in growing and non-growing cells of the human pathogen Staphylococcus aureus can be identified and even quantified by a metabolic labeling proteomic approach. S. aureus has been selected as model for this proteomic study, because it poses a major risk to our health care system by combining high pathogenicity with an increasing frequency of multiple antibiotic resistance, thus requiring the development of new anti-staphylococcal therapy strategies. Since such strategies will likely have to target extracellular and surface-exposed virulence factors as well as staphylococcal survival and adaptation capabilities, we decided to combine four subproteomic fractions: cytosolic proteins, membrane-bound proteins, cell surface-associated and extracellular proteins, to comprehensively cover the entire proteome of S. aureus. This quantitative proteomics approach integrating data ranging from gene expression to subcellular localization in growing and non growing cells is a proof of principle for whole-cell physiological proteomics that can now be extended to address physiological questions in infection-relevant settings. Importantly, with more than 1700 identified proteins (and 1450 quantified proteins) corresponding to a coverage of about three-quarters of the expressed proteins, our model study represents the most comprehensive quantification of a bacterial proteome reported to date. It thus paves the way towards a new level in understanding of cell physiology and pathophysiology of S. aureus and related pathogenic bacteria, opening new avenues for infection-related research on this crucial pathogen. PMID- 19997598 TI - Global MYCN transcription factor binding analysis in neuroblastoma reveals association with distinct E-box motifs and regions of DNA hypermethylation. AB - BACKGROUND: Neuroblastoma, a cancer derived from precursor cells of the sympathetic nervous system, is a major cause of childhood cancer related deaths. The single most important prognostic indicator of poor clinical outcome in this disease is genomic amplification of MYCN, a member of a family of oncogenic transcription factors. METHODOLOGY: We applied MYCN chromatin immunoprecipitation to microarrays (ChIP-chip) using MYCN amplified/non-amplified cell lines as well as a conditional knockdown cell line to determine the distribution of MYCN binding sites within all annotated promoter regions. CONCLUSION: Assessment of E box usage within consistently positive MYCN binding sites revealed a predominance for the CATGTG motif (p<0.0016), with significant enrichment of additional motifs CATTTG, CATCTG, CAACTG in the MYCN amplified state. For cell lines over expressing MYCN, gene ontology analysis revealed enrichment for the binding of MYCN at promoter regions of numerous molecular functional groups including DNA helicases and mRNA transcriptional regulation. In order to evaluate MYCN binding with respect to other genomic features, we determined the methylation status of all annotated CpG islands and promoter sequences using methylated DNA immunoprecipitation (MeDIP). The integration of MYCN ChIP-chip and MeDIP data revealed a highly significant positive correlation between MYCN binding and DNA hypermethylation. This association was also detected in regions of hemizygous loss, indicating that the observed association occurs on the same homologue. In summary, these findings suggest that MYCN binding occurs more commonly at CATGTG as opposed to the classic CACGTG E-box motif, and that disease associated over expression of MYCN leads to aberrant binding to additional weaker affinity E-box motifs in neuroblastoma. The co-localization of MYCN binding and DNA hypermethylation further supports the dual role of MYCN, namely that of a classical transcription factor affecting the activity of individual genes, and that of a mediator of global chromatin structure. PMID- 19997599 TI - Antibody complementarity-determining regions (CDRs): a bridge between adaptive and innate immunity. AB - BACKGROUND: It has been documented that, independently from the specificity of the native antibody (Ab) for a given antigen (Ag), complementarity determining regions (CDR)-related peptides may display differential antimicrobial, antiviral and antitumor activities. METHODOLOGY/PRINCIPAL FINDINGS: In this study we demonstrate that a synthetic peptide with sequence identical to V(H)CDR3 of a mouse monoclonal Ab (mAb) specific for difucosyl human blood group A is easily taken up by macrophages with subsequent stimulation of: i) proinflammatory cytokine production; ii) PI3K-Akt pathway and iii) TLR-4 expression. Significantly, V(H)CDR3 exerts therapeutic effect against systemic candidiasis without possessing direct candidacidal properties. CONCLUSIONS/SIGNIFICANCE: These results open a new scenario about the possibility that, beyond the half life of immunoglobulins, Ab fragments may effectively influence the antiinfective cellular immune response in a way reminiscent of regulatory peptides of innate immunity. PMID- 19997600 TI - JC virus mediates invasion and migration in colorectal metastasis. AB - INTRODUCTION: JC Virus (JCV), a human polyomavirus, is frequently present in colorectal cancers (CRCs). JCV large T-Ag (T-Ag) expressed in approximately half of all CRC's, however, its functional role in CRC is poorly understood. We hypothesized that JCV T-Ag may mediate metastasis in CRC cells through increased migration and invasion. MATERIAL AND METHODS: CRC cell lines (HCT116 and SW837) were stably transfected with JCV early transcript sequences cloned into pCR3 or empty vectors. Migration and invasion assays were performed using Boyden chambers. Global gene expression analysis was performed to identify genetic targets and pathways altered by T-Ag expression. Microarray results were validated by qRT-PCR, protein expression analyses and immunohistochemistry. Matching primary CRCs and liver metastases from 33 patients were analyzed for T Ag expression by immunohistochemistry. RESULTS: T-Ag expressing cell lines showed 2 to 3-fold increase in migration and invasion compared to controls. JCV T-Ag expression resulted in differential expression of several genetic targets, including genes that mediate cell migration and invasion. Pathway analysis suggested a significant involvement of these genes with AKT and MAPK signaling. Treatment with selective PI3K/AKT and MAPK pathway inhibitors resulted in reduced migration and invasion. In support of our in-vitro results, immunohistochemical staining of the advanced stage tumors revealed frequent JCV T-Ag expression in metastatic primary tumors (92%) as well as in their matching liver metastasis (73%). CONCLUSION: These data suggest that JCV T-Ag expression in CRC associates with a metastatic phenotype, which may partly be mediated through the AKT/MAPK signaling pathway. Frequent expression of JCV T-Ag in CRC liver metastasis provides further clues supporting a mechanistic role for JCV as a possible mediator of cellular motility and invasion in CRC. PMID- 19997601 TI - Loss of the histone pre-mRNA processing factor stem-loop binding protein in Drosophila causes genomic instability and impaired cellular proliferation. AB - BACKGROUND: Metazoan replication-dependent histone mRNAs terminate in a conserved stem-loop structure rather than a polyA tail. Formation of this unique mRNA 3' end requires Stem-loop Binding Protein (SLBP), which directly binds histone pre mRNA and stimulates 3' end processing. The 3' end stem-loop is necessary for all aspects of histone mRNA metabolism, including replication coupling, but its importance to organism fitness and genome maintenance in vivo have not been characterized. METHODOLOGY/PRINCIPAL FINDINGS: In Drosophila, disruption of the Slbp gene prevents normal histone pre-mRNA processing and causes histone pre mRNAs to utilize the canonical 3' end processing pathway, resulting in polyadenylated histone mRNAs that are no longer properly regulated. Here we show that Slbp mutants display genomic instability, including loss of heterozygosity (LOH), increased presence of chromosome breaks, tetraploidy, and changes in position effect variegation (PEV). During imaginal disc growth, Slbp mutant cells show defects in S phase and proliferate more slowly than control cells. CONCLUSIONS/SIGNIFICANCE: These data are consistent with a model in which changing the 3' end of histone mRNA disrupts normal replication-coupled histone mRNA biosynthesis and alters chromatin assembly, resulting in genomic instability, inhibition of cell proliferation, and impaired development. PMID- 19997602 TI - Simple sequence repeats and mucoid conversion: biased mucA mutagenesis in mismatch repair-deficient Pseudomonas aeruginosa. AB - In Pseudomonas aeruginosa, conversion to the mucoid phenotype marks the onset of an irreversible state of the infection in Cystic Fibrosis (CF) patients. The main pathway for mucoid conversion is mutagenesis of the mucA gene, frequently due to 1 bp deletions in a simple sequence repeat (SSR) of 5 Gs (G(5)-SSR(426)). We have recently observed that this mucA mutation is particularly accentuated in Mismatch Repair System (MRS)-deficient cells grown in vitro. Interestingly, previous reports have shown a high prevalence of hypermutable MRS-deficient strains occurring naturally in CF chronic lung infections. Here, we used mucA as a forward mutation model to systematically evaluate the role of G(5)-SSR(426) in conversion to mucoidy in a MRS-deficient background, with this being the first analysis combining SSR-dependent localized hypermutability and the acquisition of a particular virulence/persistence trait in P. aeruginosa. In this study, mucA alleles were engineered with different contents of G:C SSRs, and tested for their effect on the mucoid conversion frequency and mucA mutational spectra in a mutS deficient strain of P. aeruginosa. Importantly, deletion of G(5)-SSR(426) severely reduced the emergence frequency of mucoid variants, with no preferential site of mutagenesis within mucA. Moreover, although mutagenesis in mucA was not totally removed, this was no longer the main pathway for mucoid conversion, suggesting that G(5)-SSR(426) biased mutations towards mucA. Mutagenesis in mucA was restored by the addition of a new SSR (C(6)-SSR(431)), and even synergistically increased when G(5)-SSR(426) and C(6)-SSR(431) were present simultaneously, with the mucA mutations being restricted to -1 bp deletions within any of both G:C SSRs. These results confirm a critical role for G(5) SSR(426) enhancing the mutagenic process of mucA in MRS-deficient cells, and shed light on another mechanism, the SSR- localized hypermutability, contributing to mucoid conversion in P. aeruginosa. PMID- 19997603 TI - Adaptive vaccination strategies to mitigate pandemic influenza: Mexico as a case study. AB - BACKGROUND: We explore vaccination strategies against pandemic influenza in Mexico using an age-structured transmission model calibrated against local epidemiological data from the Spring 2009 A(H1N1) pandemic. METHODS AND FINDINGS: In the context of limited vaccine supplies, we evaluate age-targeted allocation strategies that either prioritize youngest children and persons over 65 years of age, as for seasonal influenza, or adaptively prioritize age groups based on the age patterns of hospitalization and death monitored in real-time during the early stages of the pandemic. Overall the adaptive vaccination strategy outperformed the seasonal influenza vaccination allocation strategy for a wide range of disease and vaccine coverage parameters. CONCLUSIONS: This modeling approach could inform policies for Mexico and other countries with similar demographic features and vaccine resources issues, with regard to the mitigation of the S-OIV pandemic. We also discuss logistical issues associated with the implementation of adaptive vaccination strategies in the context of past and future influenza pandemics. PMID- 19997605 TI - Association of Epstein Barr virus infection (EBV) with breast cancer in rural Indian women. AB - INTRODUCTION: Breast cancer is the most common malignancy affecting females worldwide but conventional risk factors are able to explain only a small proportion of these cases. A possible viral etiology for breast cancer has been proposed and Epstein-Barr Virus (EBV) is a widely researched candidate virus. The aim of the present study, first one of its kind from India, was to determine if there is a greater association of EBV infection with breast cancer patients as compared to patients with benign breast diseases. METHODS: We looked for expression of Epstein-Barr Virus Nuclear Antigen-1 (EBNA-1) in breast cancer tissue specimens by employing immunohistochemistry (IHC). We also measured levels of anti-EBNA-1 Immunoglobulin (IgG) antibodies in stored sera of these patients using commercial Enzyme linked Immunosorbent Assay (ELISA) kit. Patients with benign breast diseases were used as a comparison group for both immunohistochemical and serological analysis. RESULTS: 58 cases of malignant breast disease and 63 of benign breast disease (controls) were included in the study. Using manufacturer determined cut-off of 3 IU/ml, 50/55 tested (90.9%) cases and 27/33 tested (81.8%) controls were seropositive for anti-EBNA-1 IgG. Mean antibody levels were significantly higher for cases (54.22 IU/ml) as compared to controls (18.68 IU/ml). IHC for EBNA-1 was positive in 28/51 cases (54.9%). No IHC positivity was noted in the tested 30 controls. Our results show that EBNA-1 expression is seen in a significant proportion of breast cancer tissue specimens from rural India and as compared to patients with benign breast diseases these patients also have a higher immunological response against EBNA-1. PMID- 19997604 TI - REST and CoREST modulate neuronal subtype specification, maturation and maintenance. AB - BACKGROUND: The repressor element-1 silencing transcription factor/neuron restrictive silencer factor (REST/NRSF) is a master regulator of neuronal gene expression. REST functions as a modular scaffold for dynamic recruitment of epigenetic regulatory factors including its primary cofactor, the corepressor for element-1-silencing transcription factor (CoREST), to genomic loci that contain the repressor element-1 (RE1) binding motif. While REST was initially believed to silence RE1 containing neuronal genes in neural stem cells (NSCs) and non neuronal cells, emerging evidence shows an increasingly complex cell type- and developmental stage-specific repertoire of REST target genes and functions that include regulation of neuronal lineage maturation and plasticity. METHODOLOGY/PRINCIPAL FINDINGS: In this study, we utilized chromatin immunoprecipitation on chip (ChIP-chip) analysis to examine REST and CoREST functions during NSC-mediated specification of cholinergic neurons (CHOLNs), GABAergic neurons (GABANs), glutamatergic neurons (GLUTNs), and medium spiny projection neurons (MSNs). We identified largely distinct but overlapping profiles of REST and CoREST target genes during neuronal subtype specification including a disproportionately high percentage that are exclusive to each neuronal subtype. CONCLUSIONS/SIGNIFICANCE: Our findings demonstrate that the differential deployment of REST and CoREST is an important regulatory mechanism that mediates neuronal subtype specification by modulating specific gene networks responsible for inducing and maintaining neuronal subtype identity. Our observations also implicate a broad array of factors in the generation of neuronal diversity including but not limited to those that mediate homeostasis, cell cycle dynamics, cell viability, stress responses and epigenetic regulation. PMID- 19997606 TI - Comparison of trunk activity during gait initiation and walking in humans. AB - To understand the role of trunk muscles in maintenance of dynamic postural equilibrium we investigate trunk movements during gait initiation and walking, performing trunk kinematics analysis, Erector spinae muscle (ES) recordings and dynamic analysis. ES muscle expressed a metachronal descending pattern of activity during walking and gait initiation. In the frontal and horizontal planes, lateroflexion and rotation occur before in the upper trunk and after in the lower trunk. Comparison of ES muscle EMGs and trunk kinematics showed that trunk muscle activity precedes corresponding kinematics activity, indicating that the ES drive trunk movement during locomotion and thereby allowing a better pelvis mobilization. EMG data showed that ES activity anticipates propulsive phases in walking with a repetitive pattern, suggesting a programmed control by a central pattern generator. Our findings also suggest that the programs for gait initiation and walking overlap with the latter beginning before the first has ended. PMID- 19997607 TI - Apolipoprotein E-mimetics inhibit neurodegeneration and restore cognitive functions in a transgenic Drosophila model of Alzheimer's disease. AB - BACKGROUND: Mutations of the amyloid precursor protein gene (APP) are found in familial forms of Alzheimer's disease (AD) and some lead to the elevated production of amyloid-beta-protein (Abeta). While Abeta has been implicated in the causation of AD, the exact role played by Abeta and its APP precursor are still unclear. PRINCIPAL FINDINGS: In our study, Drosophila melanogaster transgenics were established as a model to analyze AD-like pathology caused by APP overexpression. We demonstrated that age related changes in the levels and pattern of synaptic proteins accompanied progressive neurodegeneration and impairment of cognitive functions in APP transgenic flies, but that these changes may be independent from the generation of Abeta. Using novel peptide mimetics of Apolipoprotein-E, COG112 or COG133 proved to be neuroprotective and significantly improved the learning and memory of APP transgenic flies. CONCLUSIONS: The development of neurodegeneration and cognitive deficits was corrected by injections of COG112 or COG133, novel mimetics of apolipoprotein-E (apoE) with neuroprotective activities. PMID- 19997608 TI - Molecular mechanism for human sperm chemotaxis mediated by progesterone. AB - Sperm chemotaxis is a chemical guiding mechanism that may orient spermatozoa to the egg surface. A picomolar concentration gradient of Progesterone (P), the main steroidal component secreted by the cumulus cells that surround the egg, attracts human spermatozoa. In order to elucidate the molecular mechanism of sperm chemotaxis mediated by P, we combine the application of different strategies: pharmacological inhibition of signaling molecules, measurements of the concentrations of second messengers and activation of the chemotactic signaling. Our data implicate a number of classic signal transduction pathways in the response and provide a model for the sequence of events, where the tmAC-cAMP-PKA pathway is activated first, followed by protein tyrosine phosphorylation (equatorial band and flagellum) and calcium mobilization (through IP(3)R and SOC channels), whereas the sGC-cGMP-PKG cascade, is activated later. These events lead to sperm orientation towards the source of the chemoattractant. The finding proposes a molecular mechanism which contributes to the understanding of the signal transduction pathway that takes place in a physiological process as chemotaxis. PMID- 19997609 TI - Effective identification of conserved pathways in biological networks using hidden Markov models. AB - BACKGROUND: The advent of various high-throughput experimental techniques for measuring molecular interactions has enabled the systematic study of biological interactions on a global scale. Since biological processes are carried out by elaborate collaborations of numerous molecules that give rise to a complex network of molecular interactions, comparative analysis of these biological networks can bring important insights into the functional organization and regulatory mechanisms of biological systems. METHODOLOGY/PRINCIPAL FINDINGS: In this paper, we present an effective framework for identifying common interaction patterns in the biological networks of different organisms based on hidden Markov models (HMMs). Given two or more networks, our method efficiently finds the top matching paths in the respective networks, where the matching paths may contain a flexible number of consecutive insertions and deletions. CONCLUSIONS/SIGNIFICANCE: Based on several protein-protein interaction (PPI) networks obtained from the Database of Interacting Proteins (DIP) and other public databases, we demonstrate that our method is able to detect biologically significant pathways that are conserved across different organisms. Our algorithm has a polynomial complexity that grows linearly with the size of the aligned paths. This enables the search for very long paths with more than 10 nodes within a few minutes on a desktop computer. The software program that implements this algorithm is available upon request from the authors. PMID- 19997610 TI - Striated muscle regulation of isometric tension by multiple equilibria. AB - Cooperative activation of striated muscle by calcium is based on the movement of tropomyosin described by the steric blocking theory of muscle contraction. Presently, the Hill model stands alone in reproducing both myosin binding data and a sigmoidal-shaped curve characteristic of calcium activation (Hill TL (1983) Two elementary models for the regulation of skeletal muscle contraction by calcium. Biophys J 44: 383-396.). However, the free myosin is assumed to be fixed by the muscle lattice and the cooperative mechanism is based on calcium-dependent interactions between nearest neighbor tropomyosin subunits, which has yet to be validated. As a result, no comprehensive model has been shown capable of fitting actual tension data from striated muscle. We show how variable free myosin is a selective advantage for activating the muscle and describe a mechanism by which a conformational change in tropomyosin propagates free myosin given constant total myosin. This mechanism requires actin, tropomyosin, and filamentous myosin but is independent of troponin. Hence, it will work equally well with striated, smooth and non-muscle contractile systems. Results of simulations with and without data are consistent with a strand of tropomyosin composed of approximately 20 subunits being moved by the concerted action of 3-5 myosin heads, which compares favorably with the predicted length of tropomyosin in the overlap region of thick and thin filaments. We demonstrate that our model fits both equilibrium myosin binding data and steady-state calcium-dependent tension data and show how both the steepness of the response and the sensitivity to calcium can be regulated by the actin-troponin interaction. The model simulates non-cooperative calcium binding both in the presence and absence of strong binding myosin as has been observed. Thus, a comprehensive model based on three well-described interactions with actin, namely, actin-troponin, actin-tropomyosin, and actin-myosin can explain the cooperative calcium activation of striated muscle. PMID- 19997611 TI - How to turn a genetic circuit into a synthetic tunable oscillator, or a bistable switch. AB - Systems and Synthetic Biology use computational models of biological pathways in order to study in silico the behaviour of biological pathways. Mathematical models allow to verify biological hypotheses and to predict new possible dynamical behaviours. Here we use the tools of non-linear analysis to understand how to change the dynamics of the genes composing a novel synthetic network recently constructed in the yeast Saccharomyces cerevisiae for In-vivo Reverse engineering and Modelling Assessment (IRMA). Guided by previous theoretical results that make the dynamics of a biological network depend on its topological properties, through the use of simulation and continuation techniques, we found that the network can be easily turned into a robust and tunable synthetic oscillator or a bistable switch. Our results provide guidelines to properly re engineering in vivo the network in order to tune its dynamics. PMID- 19997614 TI - PALM: a paralleled and integrated framework for phylogenetic inference with automatic likelihood model selectors. AB - BACKGROUND: Selecting an appropriate substitution model and deriving a tree topology for a given sequence set are essential in phylogenetic analysis. However, such time consuming, computationally intensive tasks rely on knowledge of substitution model theories and related expertise to run through all possible combinations of several separate programs. To ensure a thorough and efficient analysis and avert tedious manipulations of various programs, this work presents an intuitive framework, the phylogenetic reconstruction with automatic likelihood model selectors (PALM), with convincing, updated algorithms and a best-fit model selection mechanism for seamless phylogenetic analysis. METHODOLOGY: As an integrated framework of ClustalW, PhyML, MODELTEST, ProtTest, and several in house programs, PALM evaluates the fitness of 56 substitution models for nucleotide sequences and 112 substitution models for protein sequences with scores in various criteria. The input for PALM can be either sequences in FASTA format or a sequence alignment file in PHYLIP format. To accelerate the computing of maximum likelihood and bootstrapping, this work integrates MPICH2/PhyML, PalmMonitor and Palm job controller across several machines with multiple processors and adopts the task parallelism approach. Moreover, an intuitive and interactive web component, PalmTree, is developed for displaying and operating the output tree with options of tree rooting, branches swapping, viewing the branch length values, and viewing bootstrapping score, as well as removing nodes to restart analysis iteratively. SIGNIFICANCE: The workflow of PALM is straightforward and coherent. Via a succinct, user-friendly interface, researchers unfamiliar with phylogenetic analysis can easily use this server to submit sequences, retrieve the output, and re-submit a job based on a previous result if some sequences are to be deleted or added for phylogenetic reconstruction. PALM results in an inference of phylogenetic relationship not only by vanquishing the computation difficulty of ML methods but also providing statistic methods for model selection and bootstrapping. The proposed approach can reduce calculation time, which is particularly relevant when querying a large data set. PALM can be accessed online at http://palm.iis.sinica.edu.tw. PMID- 19997613 TI - Differing prevalence and diversity of bacterial species in fetal membranes from very preterm and term labor. AB - BACKGROUND: Intrauterine infection may play a role in preterm delivery due to spontaneous preterm labor (PTL) and preterm prolonged rupture of membranes (PPROM). Because bacteria previously associated with preterm delivery are often difficult to culture, a molecular biology approach was used to identify bacterial DNA in placenta and fetal membranes. METHODOLOGY/PRINCIPAL FINDINGS: We used broad-range 16S rDNA PCR and species-specific, real-time assays to amplify bacterial DNA from fetal membranes and placenta. 74 women were recruited to the following groups: PPROM <32 weeks (n = 26; 11 caesarean); PTL with intact membranes <32 weeks (n = 19; all vaginal birth); indicated preterm delivery <32 weeks (n = 8; all caesarean); term (n = 21; 11 caesarean). 50% (5/10) of term vaginal deliveries were positive for bacterial DNA. However, little spread was observed through tissues and species diversity was restricted. Minimal bacteria were detected in term elective section or indicated preterm deliveries. Bacterial prevalence was significantly increased in samples from PTL with intact membranes [89% (17/19) versus 50% (5/10) in term vaginal delivery p = 0.03] and PPROM (CS) [55% (6/11) versus 0% (0/11) in term elective CS, p = 0.01]. In addition, bacterial spread and diversity was greater in the preterm groups with 68% (13/19) PTL group having 3 or more positive samples and over 60% (12/19) showing two or more bacterial species (versus 20% (2/10) in term vaginal deliveries). Blood monocytes from women with PTL with intact membranes and PPROM who were 16S bacterial positive showed greater level of immune paresis (p = 0.03). A positive PCR result was associated with histological chorioamnionitis in preterm deliveries. CONCLUSION/SIGNIFICANCE: Bacteria are found in both preterm and term fetal membranes. A greater spread and diversity of bacterial species were found in tissues of women who had very preterm births. It is unclear to what extent the greater bacterial prevalence observed in all vaginal delivery groups reflects bacterial contamination or colonization of membranes during labor. Bacteria positive preterm tissues are associated with histological chorioamnionitis and a pronounced maternal immune paresis. PMID- 19997615 TI - Genome-wide identification of Schistosoma japonicum microRNAs using a deep sequencing approach. AB - BACKGROUND: Human schistosomiasis is one of the most prevalent and serious parasitic diseases worldwide. Schistosoma japonicum is one of important pathogens of this disease. MicroRNAs (miRNAs) are a large group of non-coding RNAs that play important roles in regulating gene expression and protein translation in animals. Genome-wide identification of miRNAs in a given organism is a critical step to facilitating our understanding of genome organization, genome biology, evolution, and posttranscriptional regulation. METHODOLOGY/PRINCIPAL FINDINGS: We sequenced two small RNA libraries prepared from different stages of the life cycle of S. japonicum, immature schistosomula and mature pairing adults, through a deep DNA sequencing approach, which yielded approximately 12 million high quality short sequence reads containing a total of approximately 2 million non redundant tags. Based on a bioinformatics pipeline, we identified 176 new S. japonicum miRNAs, of which some exhibited a differential pattern of expression between the two stages. Although 21 S. japonicum miRNAs are orthologs of known miRNAs within the metazoans, some nucleotides at many positions of Schistosoma miRNAs, such as miR-8, let-7, miR-10, miR-31, miR-92, miR-124, and miR-125, are indeed significantly distinct from other bilaterian orthologs. In addition, both miR-71 and some miR-2 family members in tandem are found to be clustered in a reversal direction model on two genomic loci, and two pairs of novel S. japonicum miRNAs were derived from sense and antisense DNA strands at the same genomic loci. CONCLUSIONS/SIGNIFICANCE: The collection of S. japonicum miRNAs could be used as a new platform to study the genomic structure, gene regulation and networks, evolutionary processes, development, and host-parasite interactions. Some S. japonicum miRNAs and their clusters could represent the ancestral forms of the conserved orthologues and a model for the genesis of novel miRNAs. PMID- 19997616 TI - Does haplodiploidy purge inbreeding depression in rotifer populations? AB - BACKGROUND: Inbreeding depression is an important evolutionary factor, particularly when new habitats are colonized by few individuals. Then, inbreeding depression by drift could favour the establishment of later immigrants because their hybrid offspring would enjoy higher fitness. Rotifers are the only major zooplanktonic group where information on inbreeding depression is still critically scarce, despite the fact that in cyclical parthenogenetic rotifers males are haploid and could purge deleterious recessive alleles, thereby decreasing inbreeding depression. METHODOLOGY/PRINCIPAL FINDINGS: We studied the effects of inbreeding in two populations of the cyclical parthenogenetic rotifer Brachionus plicatilis. For each population, we compared both the parental fertilization proportion and F1 fitness components from intraclonal (selfed) and interclonal (outcrossed) crosses. The parental fertilization proportion was similar for both types of crosses, suggesting that there is no mechanism to avoid selfing. In the F1 generation of both populations, we found evidence of inbreeding depression for the fitness components associated with asexual reproduction; whereas inbreeding depression was only found for one of the two sexual reproduction fitness components measured. CONCLUSIONS/SIGNIFICANCE: Our results show that rotifers, like other major zooplanktonic groups, can be affected by inbreeding depression in different stages of their life cycle. These results suggest that haplodiploidy does not purge efficiently deleterious recessive alleles. The inbreeding depression detected here has important implications when a rotifer population is founded and intraclonal crossing is likely to occur. Thus, during the foundation of new populations inbreeding depression may provide opportunities for new immigrants, increasing gene flow between populations, and affecting genetic differentiation. PMID- 19997612 TI - The severity of pandemic H1N1 influenza in the United States, from April to July 2009: a Bayesian analysis. AB - BACKGROUND: Accurate measures of the severity of pandemic (H1N1) 2009 influenza (pH1N1) are needed to assess the likely impact of an anticipated resurgence in the autumn in the Northern Hemisphere. Severity has been difficult to measure because jurisdictions with large numbers of deaths and other severe outcomes have had too many cases to assess the total number with confidence. Also, detection of severe cases may be more likely, resulting in overestimation of the severity of an average case. We sought to estimate the probabilities that symptomatic infection would lead to hospitalization, ICU admission, and death by combining data from multiple sources. METHODS AND FINDINGS: We used complementary data from two US cities: Milwaukee attempted to identify cases of medically attended infection whether or not they required hospitalization, while New York City focused on the identification of hospitalizations, intensive care admission or mechanical ventilation (hereafter, ICU), and deaths. New York data were used to estimate numerators for ICU and death, and two sources of data--medically attended cases in Milwaukee or self-reported influenza-like illness (ILI) in New York--were used to estimate ratios of symptomatic cases to hospitalizations. Combining these data with estimates of the fraction detected for each level of severity, we estimated the proportion of symptomatic patients who died (symptomatic case-fatality ratio, sCFR), required ICU (sCIR), and required hospitalization (sCHR), overall and by age category. Evidence, prior information, and associated uncertainty were analyzed in a Bayesian evidence synthesis framework. Using medically attended cases and estimates of the proportion of symptomatic cases medically attended, we estimated an sCFR of 0.048% (95% credible interval [CI] 0.026%-0.096%), sCIR of 0.239% (0.134%-0.458%), and sCHR of 1.44% (0.83%-2.64%). Using self-reported ILI, we obtained estimates approximately 7-9 x lower. sCFR and sCIR appear to be highest in persons aged 18 y and older, and lowest in children aged 5-17 y. sCHR appears to be lowest in persons aged 5-17; our data were too sparse to allow us to determine the group in which it was the highest. CONCLUSIONS: These estimates suggest that an autumn winter pandemic wave of pH1N1 with comparable severity per case could lead to a number of deaths in the range from considerably below that associated with seasonal influenza to slightly higher, but with the greatest impact in children aged 0-4 and adults 18-64. These estimates of impact depend on assumptions about total incidence of infection and would be larger if incidence of symptomatic infection were higher or shifted toward adults, if viral virulence increased, or if suboptimal treatment resulted from stress on the health care system; numbers would decrease if the total proportion of the population symptomatically infected were lower than assumed. PMID- 19997617 TI - Engineering antigen-specific T cells from genetically modified human hematopoietic stem cells in immunodeficient mice. AB - There is a desperate need for effective therapies to fight chronic viral infections. The immune response is normally fastidious at controlling the majority of viral infections and a therapeutic strategy aimed at reestablishing immune control represents a potentially powerful approach towards treating persistent viral infections. We examined the potential of genetically programming human hematopoietic stem cells to generate mature CD8+ cytotoxic T lymphocytes that express a molecularly cloned, "transgenic" human anti-HIV T cell receptor (TCR). Anti-HIV TCR transduction of human hematopoietic stem cells directed the maturation of a large population of polyfunctional, HIV-specific CD8+ cells capable of recognizing and killing viral antigen-presenting cells. Thus, through this proof-of-concept we propose that genetic engineering of human hematopoietic stem cells will allow the tailoring of effector T cell responses to fight HIV infection or other diseases that are characterized by the loss of immune control. PMID- 19997618 TI - Hepatitis C virus (HCV) genotype 1 subtype identification in new HCV drug development and future clinical practice. AB - BACKGROUND: With the development of new specific inhibitors of hepatitis C virus (HCV) enzymes and functions that may yield different antiviral responses and resistance profiles according to the HCV subtype, correct HCV genotype 1 subtype identification is mandatory in clinical trials for stratification and interpretation purposes and will likely become necessary in future clinical practice. The goal of this study was to identify the appropriate molecular tool(s) for accurate HCV genotype 1 subtype determination. METHODOLOGY/PRINCIPAL FINDINGS: A large cohort of 500 treatment-naive patients eligible for HCV drug trials and infected with either subtype 1a or 1b was studied. Methods based on the sole analysis of the 5' non-coding region (5'NCR) by sequence analysis or reverse hybridization failed to correctly identify HCV subtype 1a in 22.8%-29.5% of cases, and HCV subtype 1b in 9.5%-8.7% of cases. Natural polymorphisms at positions 107, 204 and/or 243 were responsible for mis-subtyping with these methods. A real-time PCR method using genotype- and subtype-specific primers and probes located in both the 5'NCR and the NS5B-coding region failed to correctly identify HCV genotype 1 subtype in approximately 10% of cases. The second generation line probe assay, a reverse hybridization assay that uses probes targeting both the 5'NCR and core-coding region, correctly identified HCV subtypes 1a and 1b in more than 99% of cases. CONCLUSIONS/SIGNIFICANCE: In the context of new HCV drug development, HCV genotyping methods based on the exclusive analysis of the 5'NCR should be avoided. The second-generation line probe assay is currently the best commercial assay for determination of HCV genotype 1 subtypes 1a and 1b in clinical trials and practice. PMID- 19997619 TI - Hepatitis E virus infection in central China reveals no evidence of cross-species transmission between human and swine in this area. AB - Hepatitis E virus (HEV) is a zoonotic pathogen of which several species of animal were reported as reservoirs. Swine stands out as the major reservoir for HEV infection in humans, as suggested by the close genetic relationship of swine and human virus. Since 2000, Genotype 4 HEV has become the dominant cause of hepatitis E disease in China. Recent reports showed that genotype 4 HEV is freely transmitted between humans and swine in eastern and southern China. However, the infection status of HEV in human and swine populations in central China is still unclear. This study was conducted in a rural area of central China, where there are many commercial swine farms. A total of 1476 serum and 554 fecal specimens were collected from the general human and swine populations in this area, respectively. The seroepidemiological study was conducted by enzyme-linked immunosorbent assay. Conserved genomic sequences of open reading frame 2 were detected using reverse transcription-PCR. The results indicated that the overall viral burden of the general human subjects was 0.95% (14/1476), while 7.0% (39/554) of the swine excreted HEV in stool. The positive rate of anti-HEV IgG and IgM in the serum samples was 7.9% (117/1476) and 1.6% (24/1476), respectively. Phylogenetic analysis based on the 150 nt partial sequence of the capsid protein gene showed that the 53 swine and human HEV isolates in the current study all belonged to genotype 4, clustering into three major groups. However, the HEV isolates prevalent in the human and swine populations were classified into known distinct subgenotypes, which suggested that no cross species transmission between swine and humans had taken place in this area. This result was confirmed by cloning and phylogenetic analysis of the complete capsid protein gene sequence of three representative HEV strains in the three major groups. The cross reactivity between anti-HEV IgG from human sera and the two representative strains from swine in central China was confirmed by Dot-blot assay. In conclusion, although all the HEV strains prevalent in central China belonged to genotype 4, there is no evidence of cross-species transmission between human and swine in this area. PMID- 19997620 TI - Whole-genome gene expression profiling of formalin-fixed, paraffin-embedded tissue samples. AB - BACKGROUND: We have developed a gene expression assay (Whole-Genome DASL), capable of generating whole-genome gene expression profiles from degraded samples such as formalin-fixed, paraffin-embedded (FFPE) specimens. METHODOLOGY/PRINCIPAL FINDINGS: We demonstrated a similar level of sensitivity in gene detection between matched fresh-frozen (FF) and FFPE samples, with the number and overlap of probes detected in the FFPE samples being approximately 88% and 95% of that in the corresponding FF samples, respectively; 74% of the differentially expressed probes overlapped between the FF and FFPE pairs. The WG-DASL assay is also able to detect 1.3-1.5 and 1.5-2 -fold changes in intact and FFPE samples, respectively. The dynamic range for the assay is approximately 3 logs. Comparing the WG-DASL assay with an in vitro transcription-based labeling method yielded fold-change correlations of R(2) approximately 0.83, while fold-change comparisons with quantitative RT-PCR assays yielded R(2) approximately 0.86 and R(2) approximately 0.55 for intact and FFPE samples, respectively. Additionally, the WG-DASL assay yielded high self-correlations (R(2)>0.98) with low intact RNA inputs ranging from 1 ng to 100 ng; reproducible expression profiles were also obtained with 250 pg total RNA (R(2) approximately 0.92), with approximately 71% of the probes detected in 100 ng total RNA also detected at the 250 pg level. When FFPE samples were assayed, 1 ng total RNA yielded self-correlations of R(2) approximately 0.80, while still maintaining a correlation of R(2) approximately 0.75 with standard FFPE inputs (200 ng). CONCLUSIONS/SIGNIFICANCE: Taken together, these results show that WG-DASL assay provides a reliable platform for genome-wide expression profiling in archived materials. It also possesses utility within clinical settings where only limited quantities of samples may be available (e.g. microdissected material) or when minimally invasive procedures are performed (e.g. biopsied specimens). PMID- 19997621 TI - Reactive astrocytes in glial scar attract olfactory ensheathing cells migration by secreted TNF-alpha in spinal cord lesion of rat. AB - BACKGROUND: After spinal cord injury (SCI), the formation of glial scar contributes to the failure of injured adult axons to regenerate past the lesion. Increasing evidence indicates that olfactory ensheathing cells (OECs) implanted into spinal cord are found to migrate into the lesion site and induce axons regeneration beyond glial scar and resumption of functions. However, little is known about the mechanisms of OECs migrating from injection site to glial scar/lesion site. METHODS AND FINDINGS: In the present study, we identified a link between OECs migration and reactive astrocytes in glial scar that was mediated by the tumor necrosis factor-alpha (TNF-alpha). Initially, the Boyden chamber migration assay showed that both glial scar tissue and reactive astrocyte conditioned medium promoted OECs migration in vitro. Reactive astrocyte-derived TNF-alpha and its type 1 receptor TNFR1 expressed on OECs were identified to be responsible for the promoting effect on OECs migration. TNF-alpha-induced OECs migration was demonstrated depending on activation of the extracellular signal regulated kinase (ERK) signaling cascades. Furthermore, TNF-alpha secreted by reactive astrocytes in glial scar was also showed to attract OECs migration in a spinal cord hemisection injury model of rat. CONCLUSIONS: These findings showed that TNF-alpha was released by reactive astrocytes in glial scar and attracted OECs migration by interacting with TNFR1 expressed on OECs via regulation of ERK signaling. This migration-attracting effect of reactive astrocytes on OECs may suggest a mechanism for guiding OECs migration into glial scar, which is crucial for OECs-mediated axons regrowth beyond the spinal cord lesion site. PMID- 19997622 TI - Central role of the Holliday junction helicase RuvAB in vlsE recombination and infectivity of Borrelia burgdorferi. AB - Antigenic variation plays a vital role in the pathogenesis of many infectious bacteria and protozoa including Borrelia burgdorferi, the causative agent of Lyme disease. VlsE, a 35 kDa surface-exposed lipoprotein, undergoes antigenic variation during B. burgdorferi infection of mammalian hosts, and is believed to be a critical mechanism by which the spirochetes evade immune clearance. Random, segmental recombination between the expressed vlsE gene and adjacent vls silent cassettes generates a large number of different VlsE variants within the infected host. Although the occurrence and importance of vlsE sequence variation is well established, little is known about the biological mechanism of vlsE recombination. To identify factors important in antigenic variation and vlsE recombination, we screened transposon mutants of genes known to be involved in DNA recombination and repair for their effects on infectivity and vlsE recombination. Several mutants, including those in BB0023 (ruvA), BB0022 (ruvB), BB0797 (mutS), and BB0098 (mutS-II), showed reduced infectivity in immunocompetent C3H/HeN mice. Mutants in ruvA and ruvB exhibited greatly reduced rates of vlsE recombination in C3H/HeN mice, as determined by restriction fragment polymorphism (RFLP) screening and DNA sequence analysis. In severe combined immunodeficiency (C3H/scid) mice, the ruvA mutant retained full infectivity; however, all recovered clones retained the 'parental' vlsE sequence, consistent with low rates of vlsE recombination. These results suggest that the reduced infectivity of ruvA and ruvB mutants is the result of ineffective vlsE recombination and underscores the important role that vlsE recombination plays in immune evasion. Based on functional studies in other organisms, the RuvAB complex of B. burgdorferi may promote branch migration of Holliday junctions during vlsE recombination. Our findings are consistent with those in the accompanying article by Dresser et al., and together these studies provide the first examples of trans acting factors involved in vlsE recombination. PMID- 19997624 TI - Motor vehicle crashes in diabetic patients with tight glycemic control: a population-based case control analysis. AB - BACKGROUND: Complications from diabetes mellitus can compromise a driver's ability to safely operate a motor vehicle, yet little is known about whether euglycemia predicts normal driving risks among adults with diabetes. We studied the association between glycosylated hemoglobin (HbA1c) and the risk of a motor vehicle crash using a population-based case control analysis. METHODS AND FINDINGS: We identified consecutive drivers reported to vehicle licensing authorities between January 1, 2005 to January 1, 2007 who had a diagnosis of diabetes mellitus and a HbA1c documented. The risk of a crash was calculated taking into account potential confounders including blood glucose monitoring, complications, and treatments. A total of 57 patients were involved in a crash and 738 were not involved in a crash. The mean HbA1c was lower for those in a crash than controls (7.4% versus 7.9%, unpaired t-test, p = 0.019), equal to a 26% increase in the relative risk of a crash for each 1% reduction in HbA1c (odds ratio = 1.26, 95% confidence interval 1.03-1.54). The trend was evident across the range of HbA1c values and persisted after adjustment for measured confounders (odds ratio = 1.25, 95% confidence interval 1.02-1.55). The two other significant risk factors for a crash were a history of severe hypoglycemia requiring outside assistance (odds ratio = 4.07, 95% confidence interval 2.35-7.04) and later age at diabetes diagnosis (odds ratio per decade = 1.29, 95% confidence interval 1.07 1.57). CONCLUSIONS: In this selected population, tighter glycemic control, as measured by the HbA1c, is associated with an increased risk of a motor vehicle crash. PMID- 19997623 TI - Multi-organ expression profiling uncovers a gene module in coronary artery disease involving transendothelial migration of leukocytes and LIM domain binding 2: the Stockholm Atherosclerosis Gene Expression (STAGE) study. AB - Environmental exposures filtered through the genetic make-up of each individual alter the transcriptional repertoire in organs central to metabolic homeostasis, thereby affecting arterial lipid accumulation, inflammation, and the development of coronary artery disease (CAD). The primary aim of the Stockholm Atherosclerosis Gene Expression (STAGE) study was to determine whether there are functionally associated genes (rather than individual genes) important for CAD development. To this end, two-way clustering was used on 278 transcriptional profiles of liver, skeletal muscle, and visceral fat (n = 66/tissue) and atherosclerotic and unaffected arterial wall (n = 40/tissue) isolated from CAD patients during coronary artery bypass surgery. The first step, across all mRNA signals (n = 15,042/12,621 RefSeqs/genes) in each tissue, resulted in a total of 60 tissue clusters (n = 3958 genes). In the second step (performed within tissue clusters), one atherosclerotic lesion (n = 49/48) and one visceral fat (n = 59) cluster segregated the patients into two groups that differed in the extent of coronary stenosis (P = 0.008 and P = 0.00015). The associations of these clusters with coronary atherosclerosis were validated by analyzing carotid atherosclerosis expression profiles. Remarkably, in one cluster (n = 55/54) relating to carotid stenosis (P = 0.04), 27 genes in the two clusters relating to coronary stenosis were confirmed (n = 16/17, P<10(-27 and-30)). Genes in the transendothelial migration of leukocytes (TEML) pathway were overrepresented in all three clusters, referred to as the atherosclerosis module (A-module). In a second validation step, using three independent cohorts, the A-module was found to be genetically enriched with CAD risk by 1.8-fold (P<0.004). The transcription co factor LIM domain binding 2 (LDB2) was identified as a potential high-hierarchy regulator of the A-module, a notion supported by subnetwork analysis, by cellular and lesion expression of LDB2, and by the expression of 13 TEML genes in Ldb2 deficient arterial wall. Thus, the A-module appears to be important for atherosclerosis development and, together with LDB2, merits further attention in CAD research. PMID- 19997625 TI - NMR analysis of the dynamic exchange of the NS2B cofactor between open and closed conformations of the West Nile virus NS2B-NS3 protease. AB - BACKGROUND: The two-component NS2B-NS3 proteases of West Nile and dengue viruses are essential for viral replication and established targets for drug development. In all crystal structures of the proteases to date, the NS2B cofactor is located far from the substrate binding site (open conformation) in the absence of inhibitor and lining the substrate binding site (closed conformation) in the presence of an inhibitor. METHODS: In this work, nuclear magnetic resonance (NMR) spectroscopy of isotope and spin-labeled samples of the West Nile virus protease was used to investigate the occurrence of equilibria between open and closed conformations in solution. FINDINGS: In solution, the closed form of the West Nile virus protease is the predominant conformation irrespective of the presence or absence of inhibitors. Nonetheless, dissociation of the C-terminal part of the NS2B cofactor from the NS3 protease (open conformation) occurs in both the presence and the absence of inhibitors. Low-molecular-weight inhibitors can shift the conformational exchange equilibria so that over 90% of the West Nile virus protease molecules assume the closed conformation. The West Nile virus protease differs from the dengue virus protease, where the open conformation is the predominant form in the absence of inhibitors. CONCLUSION: Partial dissociation of NS2B from NS3 has implications for the way in which the NS3 protease can be positioned with respect to the host cell membrane when NS2B is membrane associated via N- and C-terminal segments present in the polyprotein. In the case of the West Nile virus protease, discovery of low-molecular-weight inhibitors that act by breaking the association of the NS2B cofactor with the NS3 protease is impeded by the natural affinity of the cofactor to the NS3 protease. The same strategy can be more successful in the case of the dengue virus NS2B-NS3 protease. PMID- 19997626 TI - Comparative transcriptional and translational analysis of leptospiral outer membrane protein expression in response to temperature. AB - BACKGROUND: Leptospirosis is a global zoonosis affecting millions of people annually. Transcriptional changes in response to temperature were previously investigated using microarrays to identify genes potentially expressed upon host entry. Past studies found that various leptospiral outer membrane proteins are differentially expressed at different temperatures. However, our microarray studies highlighted a divergence between protein abundance and transcript levels for some proteins. Given the abundance of post-transcriptional expression control mechanisms, this finding highlighted the importance of global protein analysis systems. METHODOLOGY/PRINCIPAL FINDINGS: To complement our previous transcription study, we evaluated differences in the proteins of the leptospiral outer membrane fraction in response to temperature upshift. Outer membrane protein-enriched fractions from Leptospira interrogans grown at 30 degrees C or overnight upshift to 37 degrees C were isolated and the relative abundance of each protein was determined by iTRAQ analysis coupled with two-dimensional liquid chromatography and tandem mass spectrometry (2-DLC/MS-MS). We identified 1026 proteins with 99% confidence; 27 and 66 were present at elevated and reduced abundance respectively. Protein abundance changes were compared with transcriptional differences determined from the microarray studies. While there was some correlation between the microarray and iTRAQ data, a subset of genes that showed no differential expression by microarray was found to encode temperature regulated proteins. This set of genes is of particular interest as it is likely that regulation of their expression occurs post-transcriptionally, providing an opportunity to develop hypotheses about the molecular dynamics of the outer membrane of Leptospira in response to changing environments. CONCLUSIONS/SIGNIFICANCE: This is the first study to compare transcriptional and translational responses to temperature shift in L. interrogans. The results thus provide an insight into the mechanisms used by L. interrogans to adapt to conditions encountered in the host and to cause disease. Our results suggest down regulation of protein expression in response to temperature, and decreased expression of outer membrane proteins may facilitate minimal interaction with host immune mechanisms. PMID- 19997628 TI - Ppargamma2 is a key driver of longevity in the mouse. AB - Aging involves a progressive physiological remodeling that is controlled by both genetic and environmental factors. Many of these factors impact also on white adipose tissue (WAT), which has been shown to be a determinant of lifespan. Interrogating a transcriptional network for predicted causal regulatory interactions in a collection of mouse WAT from F2 crosses with a seed set of 60 known longevity genes, we identified a novel transcriptional subnetwork of 742 genes which represent thus-far-unknown longevity genes. Within this subnetwork, one gene was Pparg (Nr1c3), an adipose-enriched nuclear receptor previously not associated with longevity. In silico, both the PPAR signaling pathway and the transcriptional signature of Ppargamma agonist rosiglitazone overlapped with the longevity subnetwork, while in vivo, lowered expression of Pparg reduced lifespan in both the lipodystrophic Pparg1/2-hypomorphic and the Pparg2-deficient mice. These results establish Ppargamma2 as one of the determinants of longevity and suggest that lifespan may be rather determined by a purposeful genetic program than a random process. PMID- 19997627 TI - Expression of genes encoding multi-transmembrane proteins in specific primate taste cell populations. AB - BACKGROUND: Using fungiform (FG) and circumvallate (CV) taste buds isolated by laser capture microdissection and analyzed using gene arrays, we previously constructed a comprehensive database of gene expression in primates, which revealed over 2,300 taste bud-associated genes. Bioinformatics analyses identified hundreds of genes predicted to encode multi-transmembrane domain proteins with no previous association with taste function. A first step in elucidating the roles these gene products play in gustation is to identify the specific taste cell types in which they are expressed. METHODOLOGY/PRINCIPAL FINDINGS: Using double label in situ hybridization analyses, we identified seven new genes expressed in specific taste cell types, including sweet, bitter, and umami cells (TRPM5-positive), sour cells (PKD2L1-positive), as well as other taste cell populations. Transmembrane protein 44 (TMEM44), a protein with seven predicted transmembrane domains with no homology to GPCRs, is expressed in a TRPM5-negative and PKD2L1-negative population that is enriched in the bottom portion of taste buds and may represent developmentally immature taste cells. Calcium homeostasis modulator 1 (CALHM1), a component of a novel calcium channel, along with family members CALHM2 and CALHM3; multiple C2 domains; transmembrane 1 (MCTP1), a calcium-binding transmembrane protein; and anoctamin 7 (ANO7), a member of the recently identified calcium-gated chloride channel family, are all expressed in TRPM5 cells. These proteins may modulate and effect calcium signalling stemming from sweet, bitter, and umami receptor activation. Synaptic vesicle glycoprotein 2B (SV2B), a regulator of synaptic vesicle exocytosis, is expressed in PKD2L1 cells, suggesting that this taste cell population transmits tastant information to gustatory afferent nerve fibers via exocytic neurotransmitter release. CONCLUSIONS/SIGNIFICANCE: Identification of genes encoding multi-transmembrane domain proteins expressed in primate taste buds provides new insights into the processes of taste cell development, signal transduction, and information coding. Discrete taste cell populations exhibit highly specific gene expression patterns, supporting a model whereby each mature taste receptor cell is responsible for sensing, transmitting, and coding a specific taste quality. PMID- 19997629 TI - Accuracy of serological testing for the diagnosis of prevalent neurocysticercosis in outpatients with epilepsy, Eastern Cape Province, South Africa. AB - BACKGROUND: Few studies have estimated prevalence of neurocysticercosis (NCC) among persons with epilepsy in sub-Saharan Africa. While the limitations of serological testing in identification of NCC are well known, the characteristics of persons who are misdiagnosed based on serology have not been explored. The first objective of this pilot study was to estimate the prevalence of NCC in epilepsy outpatients from an area of South Africa endemic for cysticercosis. The second objective was to estimate the accuracy of serological testing in detecting NCC in these outpatients and characterize sources of disagreement between serology and neuroimaging. METHODOLOGY/PRINCIPAL FINDINGS: All out-patients aged 5 or older attending the epilepsy clinic of St. Elizabeth's Hospital in Lusikisiki, Eastern Cape Province, between July 2004 and April 2005 were invited to participate. Epidemiological data were collected by local study staff using a standardized questionnaire. Blood samples were tested by ELISA for antibody and antigen for Taenia solium. Four randomly chosen, consenting participants were transported each week to Mthatha for brain CT scan. The proportion of persons with epilepsy attending St. Elizabeth clinic with CT-confirmed NCC was 37% (95% CI: 27%-48%). Using CT as the gold standard, the sensitivity and specificity of antibody testing for identifying NCC were 54.5% (36.4%-71.9%) and 69.2% (52.4% 83.0%), respectively. Sensitivity improved to 78.6% (49.2%-95.3%) for those with active lesions. Sensitivity and specificity of antigen testing were considerably poorer. Compared to false negatives, true positives more often had active lesions. False positives were more likely to keep pigs and to have seizure onset within the past year than were true negatives. CONCLUSIONS/SIGNIFICANCE: The prevalence of NCC in South African outpatients with epilepsy is similar to that observed in other countries where cysticercosis is prevalent. Errors in classification of NCC using serology alone may reflect the natural history of NCC. PMID- 19997630 TI - The need for management capacity to achieve VISION 2020 in Sub-Saharan Africa. PMID- 19997631 TI - Temporal control of immediate early gene induction by light. AB - BACKGROUND: The light-gated cation channel channelrhodopsin-2 (ChR2) is a powerful tool for the optical induction of action potentials in neurons. Mutations of the cysteine 128 (C128) residue have been shown to greatly extend the lifetime of the conducting state of ChR2. However, until now, only subthreshold depolarizations have been reported from C128 mutants. METHODS AND FINDINGS: Here we report the induction of long high-frequency spike trains by brief light pulses in ChR2(C128A)-transfected pyramidal cells in hippocampal slice culture. ChR2(C128A)-mediated spike bursts triggered expression of the immediate early gene c-fos in pyramidal neurons. Robust and cell-specific expression of c-Fos protein was detected after a single blue light pulse and depended on action potential firing, but not on synaptic activity. However, photocurrents diminished upon repeated stimulation and limited the number of action potential bursts that could be elicited. CONCLUSIONS: We conclude that the C128A mutant is not suitable for chronic stimulation of neurons, but very useful for light-controlled induction of immediate early genes. This property of ChR2(C128A) could be harnessed to control the expression of proteins under control of the c-fos promoter with precise timing and single cell specificity. PMID- 19997632 TI - High antibody titer against apical membrane antigen-1 is required to protect against malaria in the Aotus model. AB - A Plasmodium falciparum 3D7 strain Apical Membrane Antigen-1 (AMA1) vaccine, formulated with AS02(A) adjuvant, slowed parasite growth in a recent Phase 1/2a trial, however sterile protection was not observed. We tested this AS02(A), and a Montanide ISA720 (ISA) formulation of 3D7 AMA1 in Aotus monkeys. The 3D7 parasite does not invade Aotus erythrocytes, hence two heterologous strains, FCH/4 and FVO, were used for challenge, FCH/4 AMA1 being more homologous to 3D7 than FVO AMA1. Following three vaccinations, the monkeys were challenged with 50,000 FCH/4 or 10,000 FVO parasites. Three of the six animals in the AMA+ISA group were protected against FCH/4 challenge. One monkey did not become parasitemic, another showed only a short period of low level parasitemia that self-cured, and a third animal showed a delay before exhibiting its parasitemic phase. This is the first protection shown in primates with a recombinant P. falciparum AMA1 without formulation in Freund's complete adjuvant. No animals in the AMA+AS02(A) group were protected, but this group exhibited a trend towards reduced growth rate. A second group of monkeys vaccinated with AMA+ISA vaccine was not protected against FVO challenge, suggesting strain-specificity of AMA1-based protection. Protection against FCH/4 strain correlated with the quantity of induced antibodies, as the protected animals were the only ones to have in vitro parasite growth inhibitory activity of >70% at 1:10 serum dilution; immuno-fluorescence titers >8,000; ELISA titers against full-length AMA1 >300,000 and ELISA titer against AMA1 domains1+2 >100,000. A negative correlation between log ELISA titer and day 11 cumulative parasitemia (Spearman rank r = -0.780, p value = 0.0001), further confirmed the relationship between antibody titer and protection. High titers of cross-strain inhibitory antibodies against AMA1 are therefore critical to confer solid protection, and the Aotus model can be used to down-select future AMA1 formulations, prior to advanced human trials. PMID- 19997633 TI - Distortion and signal loss in medial temporal lobe. AB - BACKGROUND: The medial temporal lobe (MTL) contains subregions that are subject to severe distortion and signal loss in functional MRI. Air/tissue and bone/tissue interfaces in the vicinity of the MTL distort the local magnetic field due to differences in magnetic susceptibility. Fast image acquisition and thin slices can reduce the amount of distortion and signal loss, but at the cost of image signal-to-noise ratio (SNR). METHODOLOGY/PRINCIPAL FINDINGS: In this paper, we quantify the severity of distortion and signal loss in MTL subregions for three different echo planar imaging (EPI) acquisitions at 3 Tesla: a conventional moderate-resolution EPI (3x3x3 mm), a conventional high-resolution EPI (1.5x1.5x2 mm), and a zoomed high-resolution EPI. We also demonstrate the advantage of reversing the phase encode direction to control the direction of distortion and to maximize efficacy of distortion compensation during data post processing. With the high-resolution zoomed acquisition, distortion is not significant and signal loss is present only in the most anterior regions of the parahippocampal gyrus. Furthermore, we find that the severity of signal loss is variable across subjects, with some subjects showing negligible loss and others showing more dramatic loss. CONCLUSIONS/SIGNIFICANCE: Although both distortion and signal loss are minimized in a zoomed field of view acquisition with thin slices, this improvement in accuracy comes at the cost of reduced SNR. We quantify this trade-off between distortion and SNR in order to provide a decision tree for design of high-resolution experiments investigating the function of subregions in MTL. PMID- 19997635 TI - An auditory illusion of infinite tempo change based on multiple temporal levels. AB - Humans and a few select insect and reptile species synchronise inter-individual behaviour without any time lag by predicting the time of future events rather than reacting to them. This is evident in music performance, dance, and drill. Although repetition of equal time intervals (i.e. isochrony) is the central principle for such prediction, this simple information is used in a flexible and complex way that accommodates both multiples, subdivisions, and gradual changes of intervals. The scope of this flexibility remains largely uncharted, and the underlying mechanisms are a matter for speculation. Here I report an auditory illusion that highlights some aspects of this behaviour and that provides a powerful tool for its future study. A sound pattern is described that affords multiple alternative and concurrent rates of recurrence (temporal levels). An algorithm that systematically controls time intervals and the relative loudness among these levels creates an illusion that the perceived rate speeds up or slows down infinitely. Human participants synchronised hand movements with their perceived rate of events, and exhibited a change in their movement rate that was several times larger than the physical change in the sound pattern. The illusion demonstrates the duality between the external signal and the internal predictive process, such that people's tendency to follow their own subjective pulse overrides the overall properties of the stimulus pattern. Furthermore, accurate synchronisation with sounds separated by more than 8 s demonstrate that multiple temporal levels are employed for facilitating temporal organisation and integration by the human brain. A number of applications of the illusion and the stimulus pattern are suggested. PMID- 19997634 TI - Seasonal differences of gene expression profiles in song sparrow (Melospiza melodia) hypothalamus in relation to territorial aggression. AB - BACKGROUND: Male song sparrows (Melospiza melodia) are territorial year-round; however, neuroendocrine responses to simulated territorial intrusion (STI) differ between breeding (spring) and non-breeding seasons (autumn). In spring, exposure to STI leads to increases in luteinizing hormone and testosterone, but not in autumn. These observations suggest that there are fundamental differences in the mechanisms driving neuroendocrine responses to STI between seasons. Microarrays, spotted with EST cDNA clones of zebra finch, were used to explore gene expression profiles in the hypothalamus after territorial aggression in two different seasons. METHODOLOGY/PRINCIPAL FINDINGS: Free-living territorial male song sparrows were exposed to either conspecific or heterospecific (control) males in an STI in spring and autumn. Behavioral data were recorded, whole hypothalami were collected, and microarray hybridizations were performed. Quantitative PCR was performed for validation. Our results show 262 cDNAs were differentially expressed between spring and autumn in the control birds. There were 173 cDNAs significantly affected by STI in autumn; however, only 67 were significantly affected by STI in spring. There were 88 cDNAs that showed significant interactions in both season and STI. CONCLUSIONS/SIGNIFICANCE: Results suggest that STI drives differential genomic responses in the hypothalamus in the spring vs. autumn. The number of cDNAs differentially expressed in relation to season was greater than in relation to social interactions, suggesting major underlying seasonal effects in the hypothalamus which may determine the differential response upon social interaction. Functional pathway analyses implicated genes that regulate thyroid hormone action and neuroplasticity as targets of this neuroendocrine regulation. PMID- 19997636 TI - The role of copy number variation in susceptibility to amyotrophic lateral sclerosis: genome-wide association study and comparison with published loci. AB - BACKGROUND: The genetic contribution to sporadic amyotrophic lateral sclerosis (ALS) has not been fully elucidated. There are increasing efforts to characterise the role of copy number variants (CNVs) in human diseases; two previous studies concluded that CNVs may influence risk of sporadic ALS, with multiple rare CNVs more important than common CNVs. A little-explored issue surrounding genome-wide CNV association studies is that of post-calling filtering and merging of raw CNV calls. We undertook simulations to define filter thresholds and considered optimal ways of merging overlapping CNV calls for association testing, taking into consideration possibly overlapping or nested, but distinct, CNVs and boundary estimation uncertainty. METHODOLOGY AND PRINCIPAL FINDINGS: In this study we screened Illumina 300K SNP genotyping data from 730 ALS cases and 789 controls for copy number variation. Following quality control filters using thresholds defined by simulation, a total of 11321 CNV calls were made across 575 cases and 621 controls. Using region-based and gene-based association analyses, we identified several loci showing nominally significant association. However, the choice of criteria for combining calls for association testing has an impact on the ranking of the results by their significance. Several loci which were previously reported as being associated with ALS were identified here. However, of another 15 genes previously reported as exhibiting ALS-specific copy number variation, only four exhibited copy number variation in this study. Potentially interesting novel loci, including EEF1D, a translation elongation factor involved in the delivery of aminoacyl tRNAs to the ribosome (a process which has previously been implicated in genetic studies of spinal muscular atrophy) were identified but must be treated with caution due to concerns surrounding genomic location and platform suitability. CONCLUSIONS AND SIGNIFICANCE: Interpretation of CNV association findings must take into account the effects of filtering and combining CNV calls when based on early genome-wide genotyping platforms and modest study sizes. PMID- 19997637 TI - Susceptibility of human melanoma cells to autologous natural killer (NK) cell killing: HLA-related effector mechanisms and role of unlicensed NK cells. AB - BACKGROUND: Despite Natural Killer (NK) cells were originally defined as effectors of spontaneous cytotoxicity against tumors, extremely limited information is so far available in humans on their capability of killing cancer cells in an autologous setting. METHODOLOGY/PRINCIPAL FINDINGS: We have established a series of primary melanoma cell lines from surgically resected specimens and here showed that human melanoma cells were highly susceptible to lysis by activated autologous NK cells. A variety of NK cell activating receptors were involved in killing: particularly, DNAM-1 and NKp46 were the most frequently involved. Since self HLA class I molecules normally play a protective role from NK cell-mediated attack, we analyzed HLA class I expression on melanomas in comparison to autologous lymphocytes. We found that melanoma cells presented specific allelic losses in 50% of the patients analyzed. In addition, CD107a degranulation assays applied to NK cells expressing a single inhibitory receptor, revealed that, even when expressed, specific HLA class I molecules are present on melanoma cell surface in amount often insufficient to inhibit NK cell cytotoxicity. Remarkably, upon activation, also the so called "unlicensed" NK cells, i.e. NK cells not expressing inhibitory receptor specific for self HLA class I molecules, acquired the capability of efficiently killing autologous melanoma cells, thus additionally contributing to the lysis by a mechanism independent of HLA class I expression on melanoma cells. CONCLUSIONS/SIGNIFICANCE: We have investigated in details the mechanisms controlling the recognition and lysis of melanoma cells by autologous NK cells. In these autologous settings, we demonstrated an efficient in vitro killing upon NK cell activation by mechanisms that may be related or not to abnormalities of HLA class I expression on melanoma cells. These findings should be taken into account in the design of novel immunotherapy approaches against melanoma. PMID- 19997638 TI - Comparative effects of heterologous TRPV1 and TRPM8 expression in rat hippocampal neurons. AB - Heterologous channel expression can be used to control activity in select neuronal populations, thus expanding the tools available to modern neuroscience. However, the secondary effects of exogenous channel expression are often left unexplored. We expressed two transient receptor potential (TRP) channel family members, TRPV1 and TRPM8, in cultured hippocampal neurons. We compared functional expression levels and secondary effects of channel expression and activation on neuronal survival and signaling. We found that activation of both channels with appropriate agonist caused large depolarizing currents in voltage-clamped hippocampal neurons, exceeding the amplitude responses to a calibrating 30 mM KCl stimulation. Both TRPV1 and TRPM8 currents were reduced but not eliminated by 4 hr incubation in saturating agonist concentration. In the case of TRPV1, but not TRPM8, prolonged agonist exposure caused strong calcium-dependent toxicity. In addition, TRPV1 expression depressed synaptic transmission dramatically without overt signs of toxicity, possibly due to low-level TRPV1 activation in the absence of exogenous agonist application. Despite evidence of expression at presynaptic sites, in addition to somatodendritic sites, TRPM8 expression alone exhibited no effects on synaptic transmission. Therefore, by a number of criteria, TRPM8 proved the superior choice for control over neuronal membrane potential. This study also highlights the need to explore potential secondary effects of long-term expression and activation of heterologously introduced channels. PMID- 19997639 TI - Red-mediated transposition and final release of the mini-F vector of a cloned infectious herpesvirus genome. AB - Bacterial artificial chromosomes (BACs) are well-established cloning vehicles for functional genomics and for constructing targeting vectors and infectious viral DNA clones. Red-recombination-based mutagenesis techniques have enabled the manipulation of BACs in Escherichia coli without any remaining operational sequences. Here, we describe that the F-factor-derived vector sequences can be inserted into a novel position and seamlessly removed from the present location of the BAC-cloned DNA via synchronous Red-recombination in E. coli in an en passant mutagenesis-based procedure. Using this technique, the mini-F elements of a cloned infectious varicella zoster virus (VZV) genome were specifically transposed into novel positions distributed over the viral DNA to generate six different BAC variants. In comparison to the other constructs, a BAC variant with mini-F sequences directly inserted into the junction of the genomic termini resulted in highly efficient viral DNA replication-mediated spontaneous vector excision upon virus reconstitution in transfected VZV-permissive eukaryotic cells. Moreover, the derived vector-free recombinant progeny exhibited virtually indistinguishable genome properties and replication kinetics to the wild-type virus. Thus, a sequence-independent, efficient, and easy-to-apply mini-F vector transposition procedure eliminates the last hurdle to perform virtually any kind of imaginable targeted BAC modifications in E. coli. The herpesviral terminal genomic junction was identified as an optimal mini-F vector integration site for the construction of an infectious BAC, which allows the rapid generation of mutant virus without any unwanted secondary genome alterations. The novel mini-F transposition technique can be a valuable tool to optimize, repair or restructure other established BACs as well and may facilitate the development of gene therapy or vaccine vectors. PMID- 19997640 TI - Optimizing odor-baited trap methods for collecting mosquitoes during the malaria season in The Gambia. AB - BACKGROUND: Baited traps are potential tools for removal or surveillance of disease vectors. To optimize the use of counter-flow traps baited with human odor (nylon socks that had been worn for a single day) to capture wild mosquitoes in the Gambia, investigations were conducted at a field experimental site. METHODOLOGY/PRINCIPAL FINDINGS: Experiments employing Latin square design were conducted with a set of six huts to investigate the effects of the following on overnight mosquito trap catches: (1) placement of traps indoors or immediately outdoors, CO(2) supply, and presence of a human subject in the hut; (2) trap height for collecting mosquitoes immediately outdoors; (3) height and distance from hut; (4) interaction between multiple traps around a single hut and entry of mosquitoes into huts. A total of 106,600 adult mosquitoes (9.1% Anopheles gambiae s.l., 4.0% other Anopheles species) were collected over 42 nights. The high numbers of An. gambiae s.l. and other mosquitoes collected by odor-baited traps required CO(2) but were largely independent of the presence of a person sleeping in the hut or of trap placement indoors or outdoors. For outdoor collection that is considered less intrusive, traps opening 15 cm above the floor of the hut veranda were more highly effective than traps at other heights or further from the hut. There was no significant evidence of saturation or competition by the traps, with multiple traps around a hut each collecting almost as many mosquitoes as single traps and no effect on the numbers of mosquitoes entering the huts. CONCLUSIONS/SIGNIFICANCE: The outdoor trapping protocol is convenient to compare attractiveness of different odors or synthetic chemicals to malaria vectors and other wild mosquitoes. The finding that such traps are reliably attractive in the presence or absence of a human volunteer encourages their potential development as standardised surveillance tools. PMID- 19997641 TI - Distinct roles for ROCK1 and ROCK2 in the regulation of keratinocyte differentiation. AB - BACKGROUND: The human epidermis is comprised of several layers of specialized epithelial cells called keratinocytes. Normal homoeostasis of the epidermis requires that the balance between keratinocyte proliferation and terminal differentiation be tightly regulated. The mammalian serine/threonine kinases (ROCK1 and ROCK2) are well-characterised downstream effectors of the small GTPase RhoA. We have previously demonstrated that the RhoA/ROCK signalling pathway plays an important role in regulation of human keratinocyte proliferation and terminal differentiation. In this paper we addressed the question of which ROCK isoform was involved in regulation of keratinocyte differentiation. METHODOLOGY AND PRINCIPAL FINDINGS: We used RNAi to specifically knockdown ROCK1 or ROCK2 expression in cultured human keratinocytes. ROCK1 depletion results in decreased keratinocyte adhesion to fibronectin and an increase in terminal differentiation. Conversely, ROCK2 depletion results in increased keratinocyte adhesion to fibronectin and inhibits terminal differentiation. CONCLUSION: These data suggest that ROCK1 and ROCK2 play distinct roles in regulating keratinocyte adhesion and terminal differentiation. PMID- 19997642 TI - Comprehensive analysis of inflammatory immune mediators in vitreoretinal diseases. AB - Inflammation affects the formation and the progression of various vitreoretinal diseases. We performed a comprehensive analysis of inflammatory immune mediators in the vitreous fluids from total of 345 patients with diabetic macular edema (DME, n = 92), proliferative diabetic retinopathy (PDR, n = 147), branch retinal vein occlusion (BRVO, n = 30), central retinal vein occlusion (CRVO, n = 13) and rhegmatogenous retinal detachment (RRD, n = 63). As a control, we selected a total of 83 patients with either idiopathic macular hole (MH) or idiopathic epiretinal membrane (ERM) that were free of major pathogenic intraocular changes, such as ischemic retina and proliferative membranes. The concentrations of 20 soluble factors (nine cytokines, six chemokines, and five growth factors) were measured simultaneously by multiplex bead analysis system. Out of 20 soluble factors, three factors: interleukin-6 (IL-6), interleukin-8 (IL-8), and monocyte chemoattractant protein-1 (MCP-1) were significantly elevated in all groups of vitreoretinal diseases (DME, PDR, BRVO, CRVO, and RRD) compared with control group. According to the correlation analysis in the individual patient's level, these three factors that were simultaneously increased, did not show any independent upregulation in all the examined diseases. Vascular endothelial growth factor (VEGF) was significantly elevated in patients with PDR and CRVO. In PDR patients, the elevation of VEGF was significantly correlated with the three factors: IL-6, IL-8, and MCP-1, while no significant correlation was observed in CRVO patients. In conclusion, multiplex bead system enabled a comprehensive soluble factor analysis in vitreous fluid derived from variety of patients. Major three factors: IL-6, IL-8, and MCP-1 were strongly correlated with each other indicating a common pathway involved in inflammation process in vitreoretinal diseases. PMID- 19997643 TI - Regional gene expression of LOX-1, VCAM-1, and ICAM-1 in aorta of HIV-1 transgenic rats. AB - BACKGROUND: Increased prevalence of atherosclerotic cardiovascular disease in HIV infected patients has been observed. The cause of this accelerated atherosclerosis is a matter of controversy. As clinical studies are complicated by a multiplicity of risk-factors and a low incidence of hard endpoints, studies in animal models could be attractive alternatives. METHODOLOGY/PRINCIPAL FINDINGS: We evaluated gene expression of lectin-like oxidized-low-density lipoprotein receptor-1 (LOX-1), vascular cell adhesion molecule-1 (VCAM-1), and intercellular adhesion molecule-1 (ICAM-1) in HIV-1 transgenic (HIV-1Tg) rats; these genes are all thought to play important roles in early atherogenesis. Furthermore, the plasma level of sICAM-1 was measured. We found that gene expressions of LOX-1 and VCAM-1 were higher in the aortic arch of HIV-1Tg rats compared to controls. Also, the level of sICAM-1 was elevated in the HIV-1Tg rats compared to controls, but the ICAM-1 gene expression profile did not show any differences between the groups. CONCLUSIONS/SIGNIFICANCE: HIV-1Tg rats have gene expression patterns indicating endothelial dysfunction and accelerated atherosclerosis in aorta, suggesting that HIV-infection per se may cause atherosclerosis. This transgenic rat model may be a very promising model for further studies of the pathophysiology behind HIV-associated cardiovascular disease. PMID- 19997644 TI - Protective effects of human iPS-derived retinal pigment epithelium cell transplantation in the retinal dystrophic rat. AB - Transformation of somatic cells with a set of embryonic transcription factors produces cells with the pluripotent properties of embryonic stem cells (ESCs). These induced pluripotent stem (iPS) cells have the potential to differentiate into any cell type, making them a potential source from which to produce cells as a therapeutic platform for the treatment of a wide range of diseases. In many forms of human retinal disease, including age-related macular degeneration (AMD), the underlying pathogenesis resides within the support cells of the retina, the retinal pigment epithelium (RPE). As a monolayer of cells critical to photoreceptor function and survival, the RPE is an ideally accessible target for cellular therapy. Here we report the differentiation of human iPS cells into RPE. We found that differentiated iPS-RPE cells were morphologically similar to, and expressed numerous markers of developing and mature RPE cells. iPS-RPE are capable of phagocytosing photoreceptor material, in vitro and in vivo following transplantation into the Royal College of Surgeons (RCS) dystrophic rat. Our results demonstrate that iPS cells can be differentiated into functional iPS-RPE and that transplantation of these cells can facilitate the short-term maintenance of photoreceptors through phagocytosis of photoreceptor outer segments. Long-term visual function is maintained in this model of retinal disease even though the xenografted cells are eventually lost, suggesting a secondary protective host cellular response. These findings have identified an alternative source of replacement tissue for use in human retinal cellular therapies, and provide a new in vitro cellular model system in which to study RPE diseases affecting human patients. PMID- 19997646 TI - A cost of sexual attractiveness to high-fitness females. AB - Adaptive mate choice by females is an important component of sexual selection in many species. The evolutionary consequences of male mate preferences, however, have received relatively little study, especially in the context of sexual conflict, where males often harm their mates. Here, we describe a new and counterintuitive cost of sexual selection in species with both male mate preference and sexual conflict via antagonistic male persistence: male mate choice for high-fecundity females leads to a diminished rate of adaptive evolution by reducing the advantage to females of expressing beneficial genetic variation. We then use a Drosophila melanogaster model system to experimentally test the key prediction of this theoretical cost: that antagonistic male persistence is directed toward, and harms, intrinsically higher-fitness females more than it does intrinsically lower-fitness females. This asymmetry in male persistence causes the tails of the population's fitness distribution to regress towards the mean, thereby reducing the efficacy of natural selection. We conclude that adaptive male mate choice can lead to an important, yet unappreciated, cost of sex and sexual selection. PMID- 19997645 TI - CS-SELEX generates high-affinity ssDNA aptamers as molecular probes for hepatitis C virus envelope glycoprotein E2. AB - Currently, the development of effective diagnostic reagents as well as treatments against Hepatitis C virus (HCV) remains a high priority. In this study, we have described the development of an alive cell surface--Systematic Evolution of Ligands by Exponential Enrichment (CS-SELEX) technique and screened the functional ssDNA aptamers that specifically bound to HCV envelope surface glycoprotein E2. Through 13 rounds of selection, the CS-SELEX generated high affinity ssDNA aptamers, and the selected ssDNA aptamer ZE2 demonstrated the highest specificity and affinity to E2-positive cells. HCV particles could be specifically captured and diagnosed using the aptamer ZE2. A good correlation was observed in HCV patients between HCV E2 antigen-aptamer assay and assays for HCV RNA quantities or HCV antibody detection. Moreover, the selected aptamers, especially ZE2, could competitively inhibit E2 protein binding to CD81, an important HCV receptor, and significantly block HCV cell culture (HCVcc) infection of human hepatocytes (Huh7.5.1) in vitro. Our data demonstrate that the newly selected ssDNA aptamers, especially aptamer ZE2, hold great promise for developing new molecular probes, as an early diagnostic reagent for HCV surface antigen, or a therapeutic drug specifically for HCV. PMID- 19997647 TI - Deep brain stimulation reveals emotional impact processing in ventromedial prefrontal cortex. AB - We tested the hypothesis that modulation of monoaminergic tone with deep-brain stimulation (DBS) of subthalamic nucleus would reveal a site of reactivity in the ventromedial prefrontal cortex that we previously identified by modulating serotonergic and noradrenergic mechanisms by blocking serotonin-noradrenaline reuptake sites. We tested the hypothesis in patients with Parkinson's disease in whom we had measured the changes of blood flow everywhere in the brain associated with the deep brain stimulation of the subthalamic nucleus. We determined the emotional reactivity of the patients as the average impact of emotive images rated by the patients off the DBS. We then searched for sites in the brain that had significant correlation of the changes of blood flow with the emotional impact rated by the patients. The results indicate a significant link between the emotional impact when patients are not stimulated and the change of blood flow associated with the DBS. In subjects with a low emotional impact, activity measured as blood flow rose when the electrode was turned on, while in subjects of high impact, the activity at this site in the ventromedial prefrontal cortex declined when the electrode was turned on. We conclude that changes of neurotransmission in the ventromedial prefrontal cortex had an effect on the tissue that depends on changes of monoamine concentration interacting with specific combinations of inhibitory and excitatory monoamine receptors. PMID- 19997648 TI - Alternation of sound location induces visual motion perception of a static object. AB - BACKGROUND: Audition provides important cues with regard to stimulus motion although vision may provide the most salient information. It has been reported that a sound of fixed intensity tends to be judged as decreasing in intensity after adaptation to looming visual stimuli or as increasing in intensity after adaptation to receding visual stimuli. This audiovisual interaction in motion aftereffects indicates that there are multimodal contributions to motion perception at early levels of sensory processing. However, there has been no report that sounds can induce the perception of visual motion. METHODOLOGY/PRINCIPAL FINDINGS: A visual stimulus blinking at a fixed location was perceived to be moving laterally when the flash onset was synchronized to an alternating left-right sound source. This illusory visual motion was strengthened with an increasing retinal eccentricity (2.5 deg to 20 deg) and occurred more frequently when the onsets of the audio and visual stimuli were synchronized. CONCLUSIONS/SIGNIFICANCE: We clearly demonstrated that the alternation of sound location induces illusory visual motion when vision cannot provide accurate spatial information. The present findings strongly suggest that the neural representations of auditory and visual motion processing can bias each other, which yields the best estimates of external events in a complementary manner. PMID- 19997649 TI - The impact of the Wenchuan earthquake on birth outcomes. AB - BACKGROUND: Earthquakes and other catastrophic events frequently occurring worldwide can be considered as outliers and cause a growing and urgent need to improve our understanding of the negative effects imposed by such disasters. Earthquakes can intensively impact the birth outcomes upon psychological and morphological development of the unborn children, albeit detailed characteristics remain obscure. METHODS AND FINDINGS: We utilized the birth records at Du Jiang Yan and Peng Zhou counties to investigate the birth outcomes as a consequence of a major earthquake occurred in Wenchuan, China on May 12, 2008. Totally 13,003 of neonates were recorded, with 6638 and 6365 for pre- and post- earthquake, respectively. Significant low birthweight, high ratio of low birthweight, and low Apgar scores of post-earthquake group were observed. In contrast, the sex ratio at birth, birth length and length of gestation did not show statistical differences. The overall ratio of birth-defect in the post-earthquake (1.18%) is statistically high than that of pre-earthquake (0.99%), especially for those in the first trimester on earthquake day (1.47%). The birth-defect spectrum was dramatically altered after earthquake, with the markedly increased occurrences of ear malformations. The ratio of preterm birth post-earthquake (7.41%) is significant increased than that of pre-earthquake (5.63%). For the birth outcomes of twins, significant differences of the ratio of twins, birth weight, ratio of low birthweight and birth-defect rate were observed after earthquake. CONCLUSION: A hospital-based study of birth outcomes impacted by the Wenchuan earthquake shows that the earthquake was associated with significant effects on birth outcomes, indicating it is a major monitor for long-term pregnant outcomes. PMID- 19997650 TI - Two Gln187 mutants of human soluble APRIL inhibit proliferation of lung carcinoma A549 cells. AB - Soluble APRIL (sAPRIL), the active form of a proliferation-inducing ligand (APRIL), is implicated in the proliferation of tumor cells. Suppressing APRIL function has been considered as a potential strategy for the therapy of APRIL associated tumors. In the present study, we generated human sAPRIL and its two mutants, Gln187-D-sAPRIL (Gln187 deleted) and Gly187-sAPRIL (Gln187 replaced by Gly). In vitro experiments showed that the two mutants had similar specific binding capacity to lung carcinoma A549 cells compared to the wild-type sAPRIL, and both, especially Gly187-sAPRIL, exhibited significant antagonistic effect on sAPRIL-induced tumor cell proliferation in a dose-dependent manner, which might be predominantly mediated by blocking sAPRIL-induced MEK and ERK phosphorylation but not p38MAPK or JNK signaling. In vivo experiments with nude mice bearing A549 cell-derived xenograft tumor showed that only the Gly187-sAPRIL mutant could significantly suppress the tumor growth. These results suggest that Gln187 may be a crucial amino acid in APRIL-mediated tumor cell proliferation via the MEK-ERK signaling pathway and that the sAPRIL mutants may serve as novel potential antagonists of APRIL for the therapy of APRIL-associated cancers. PMID- 19997651 TI - Molecular cloning, expression and characterization of Bmserpin-2 gene from Bombyx mori. AB - Serpins are a broadly distributed family of protease inhibitors. In this study, the gene encoding Bombyx mori serpin-2 (Bmserpin-2) was cloned and expressed in E. coli. The Bmserpin-2 cDNA contains a 1125 bp open reading frame (ORF). The deduced protein has 374 amino-acid residues, contains a conserved SERPIN domain and shares extensive homology with other invertebrate serpins. RT-PCR analysis showed that Bmserpin-2 was expressed in all developmental stages of B. mori larvae and various larval tissues. Subcellular localization analysis indicated that Bmserpin-2 protein was located in the cytoplasm. Interestingly, real-time quantitative PCR revealed that the expression of Bmserpin-2 in the midgut of susceptible B. mori strain 306 significantly increased at 72 hours post inoculation (hpi) when infected with BmNPV. However, there was no significant increase of the Bmserpin-2 expression in resistant strain NB infected with BmNPV. Thus, our data indicates that Bmserpin-2 may be involved in B. mori antiviral response. PMID- 19997652 TI - New bradykinin B(2) receptor antagonists - influence of C-terminal segment modifications on their pharmacological properties. AB - In the present study we describe the synthesis and some pharmacological properties of eight new analogues of bradykinin (BK). Two peptides were designed by substitution of position 7 or 8 of the known [D-Arg(0),Hyp(3),Thi(5,8),D Phe(7)]BK antagonist (Stewart's antagonist) with L-pipecolic acid (L-Pip). The next two analogues were obtained by replacement of the d-Phe residue in position 7 of the Stewart's peptide with L-beta(2)-isoproline (L-beta(2)-iPro) or L beta(3)-homoproline (L-beta(3)-hPro). The four analogues mentioned above were also prepared in N-acylated form with 1-adamantaneacetic acid (Aaa). Biological activity of the compounds was assessed by isolated rat uterus and rat blood pressure tests. Our results showed that L-Pip in position 7 slightly increased antagonistic potency in the blood pressure test, but it turned the analogue into an agonist in the rat uterus test. Replacement of Thi by L-Pip in position 8 also enhanced antagonism in the rat pressure test but preserved the antagonism in the rat uterus test. L-beta(2)-iPro or L-beta(3)-hPro in position 7 decreased the potencies in both tests. We also demonstrated that acylation of the N-terminus did not increase, as was claimed previously, the antagonistic potencies of the resulting peptides. The results thus support the hypothesis about the existence of different types of BK receptors in the rat uterus and blood vessels. Our studies provide new information about the structure-activity relationship of BK antagonists which may help in designing more potent BK receptor blockers. PMID- 19997653 TI - Influence of megestrol acetate on nutrition and inflammation in dialysis patients - preliminary results. AB - Malnutrition is a common clinical problem in dialysis patients. So far the management of malnutrition in this population has not been fully successful. The aim of the study was to evaluate the efficacy and safety of use of megestrol acetate suspension in malnourished dialysis patients. Twenty-six hypoalbuminemic (albumin < or = 3.8 g/dl) dialysis patients took 160 mg of megestrol acetate daily for a period of two months. Anthropometry (dry weight, body mass index) and biochemical measurements of nutrition (serum albumin, triglycerides, total cholesterol) and inflammation (hsCRP, IL-1beta, IL-6) were performed on a monthly basis. The treatment led to a statistically significant increase (P < 0.05) in anthropometry and albumin concentration, with no statistically significant changes in total cholesterol, triglycerides and indices of inflammation. Side effects included overhydration, diarrhoea and hyperglycaemia. Thus, megestrol acetate may be an effective therapeutic agent in improving the nutritional status of carefully selected dialysis patients, while it might not mitigate inflammation. Because of the prevalent side effects it must be monitored closely. PMID- 19997654 TI - Matrix metalloproteinases in serum of Emery-Dreifuss muscular dystrophy patients. AB - In the pathogenesis of dilated cardiomyopathy (DCM) in Emery-Dreifuss muscular dystrophy (EDMD) matrix metalloproteinases (MMPs) are supposed to be involved and may have diagnostic/prognostic value. Serum levels of MT1-MMP, MMP-2 and MMP-9 were quantified by ELISA and zymography in 22 EDMD patients and 15 age-matched controls. In the autosomal-dominant EDMD MMP-2 and MT1-MMP were increased in all cases, and MMP-9 was increased in two of the eight examined patients. In the X linked EDMD MMP-2 expression was increased in all the cases, MMP-9 level was elevated in 3 of the 14 cases, and MT1-MMP was decreased in eight of these patients. There was no evident correlation between the MMPs level and the different cardiac parameters including left-ventricular end-diastolic diameter, left atrial diameter and left ventricular ejection fraction in either form of EDMD. The presented results indicate that a changed level of matrix metalloproteinases, especially that of MMP-2 in serum, may be of value for detection of cardiac involvement in EDMD patients, especially in those patients with no evident subjective cardiac symptoms. Further follow-up studies of MMPs are needed to check if their determination is of value for monitoring of the progression of atrial/ventricular dilatation. MMPs determinations may also be useful for monitoring DCM treatment by synthetic MMPs inhibitors. PMID- 19997655 TI - Elevated advanced oxidation protein products levels in patients with liver cirrhosis. AB - Serum concentrations of advanced oxidation protein products (AOPPs) and glycation end products (AGEs) were assessed with respect to functional compromise of liver, as determined by the Child-Pugh and MELD scores. Patients with decompensated liver cirrhosis (Child-Pugh B and C) exhibited significantly higher serum concentrations of AOPPs than both patients with compensated liver cirrhosis (Child-Pugh A) and controls. The levels of plasma AGEs in all liver cirrhotic patients were higher when compared with those with the controls and this difference was statistically significant. Plasma total antioxidant status of the patients was significantly lower than that of controls. Significant positive correlations between AOPPs level and the MELD score and between the oxidative stress index and the MELD score were found in all patients with liver cirrhosis. Altered AOPPs levels in decompensated patients may influence the potency of oxidative stress and the progression of liver disease. PMID- 19997656 TI - Full length genome sequence of Polish isolate of Beet soil-borne virus confirms low level of genetic diversity. AB - The complete nucleotide sequence of a Polish isolate of Beet soil-borne virus was determined for the first time. The genome organization was identical with those previously established for isolates from Germany and China. A comparison of the Polish isolate with others deposited in GenBank revealed high level of nucleotide identity, about 98-100%, throughout the genome analyzed. The ratio between non synonymous and synonymous substitutions was rather low suggesting a negative selective pressure. The non-synonymous mutations were particulary frequent in triple gene block. PMID- 19997657 TI - Small intestine bacterial overgrowth is frequent in cystic fibrosis: combined hydrogen and methane measurements are required for its detection. AB - INTRODUCTION: Hydrogen breath test (BT) is commonly used as a diagnostic tool for the detection of small intestine bacterial overgrowth (SIBO). It was reported that colonic methane production is far more frequent in cystic fibrosis (CF) patients than in other subjects. Therefore, measuring exclusively hydrogen in the diagnostic breath test for diagnosing SIBO might be of limited value. We aimed to assess the usefulness of combined measurement of hydrogen and methane expiration for the diagnosis of SIBO in CF. MATERIAL AND METHODS: The study comprised 62 CF patients aged 5 to 18 years. Three-hundred-ninety subjects assessed due to gastrointestinal symptoms for the presence of SIBO served as a comparative group. In all subjects hydrogen/methane BT using glucose was performed. A positive BT was defined as fasting hydrogen > or = 20 ppm or fasting methane > or = 10 ppm or a rise of > or = 12 ppm hydrogen or > or = 6 ppm methane over baseline during the test. RESULTS: In 23 (37.1%) CF patients and in 52 (13.3%) subjects from the comparative group abnormal BT results were found. In seven (11.3%) CF patients and 29 (7.4%) of the other subjects studied methane measurement allowed diagnosis of SIBO. CONCLUSIONS: Small intestine bacterial overgrowth is frequent in cystic fibrosis. For its detection in cystic fibrosis and other gastrointestinal patients, combined hydrogen and methane measurement instead of hydrogen breath test should be applied. Without the additional measurement of methane a significant percentage of SIBO will be missed. PMID- 19997664 TI - Health informatics in Japan, update, 2008- 2009. PMID- 19997658 TI - Measurement of functional independence level and falls-risk in individuals with undiagnosed phenylketonuria. AB - The aim of the study was to determine the level of functional independence in adult patients with previously undiagnosed or untreated phenylketonuria (PKU). The study was conducted among 400 intellectually impaired adult residents of Social Welfare Homes in South-Eastern Poland born prior to the introduction of neonatal PKU screening programs. PKU was screened by filter paper test using tandem mass spectrometry methods, and confirmed by gas chromatography-mass spectrometric analysis of PKU organic acids in urine. Degree of functional independence included the assessment of activities of daily living (Barthel Index) and measures of balance and gait (Tinetti scale). Eleven individuals with previously untreated PKU were identified whereby eight presented with moderate disability and three with mild disability. Six had a high risk of falls and five had a moderate risk of falls. This study indicates that there is considerable number of undiagnosed PKU patients within the Polish population who require assessment and management in order to reduce the impact of the neurological and neuropsychiatric problems associated with the condition. Appropriate therapy for those with undiagnosed PKU should, in particular, address the risk of falls. PMID- 19997665 TI - Risk of simultaneous phenotypic expression of hip and elbow dysplasia in dogs: a study of 1,411 radiographic examinations sent for official scoring. AB - In order to look for phenotypic correlation between hip (HD) and elbow (ED) dysplasia, we used radiographic scoring obtained from 1,411 dogs of different breeds, which were evaluated for authoritative grading of both conditions. In this population, we found that the risk ratio for an animal to be simultaneously affected by HD and ED is 1.67. For a dog with ED, the risk ratio to be affected by HD increases as the ED grade increases. Similarly, for a dog affected by HD, the risk ratio to be affected by ED increases as the HD grade increases. In a dog affected by HD or ED, the clinician should look for the second condition in the same animal. Due to the low, yet positive correlation, selection against one trait will not affect the other trait sufficiently. Therefore selection has to be conducted at reduction of HD as well as reduction of ED. PMID- 19997666 TI - The effects of ovariectomy on bone mineral density, geometrical, and biomechanical characteristics in the rabbit femur. AB - The aim of the study was to evaluate the bone mineral density, as well as the biomechanic and morphometric changes in the femur of ovariectomised rabbits. Twenty-four six-month-old New Zealand rabbits were randomly divided into an ovariectomy (n = 12) and a sham (n = 12) group. Six rabbits in each group were euthanatized at eight and 16 weeks after surgery, and the femora were resected. The morphometric data were obtained from tomographic images. Periosteal and endosteal diameters and cortical thickness were measured. Total cross- sectional, cortical and medullary areas were also measured. The bone mineral content, the bone area and the bone mineral density were measured from the proximal, distal and midshaft of the femur as well as the total femur by dual energy X-ray absorptiometry. Employing the three-point bending method, the ultimate force, stiffness and work-to-failure were measured. The mechanical data were normalised to obtain intrinsic biomechanical properties such as ultimate stress, elastic modulus, and toughness, all of which are independent of size and shape. The results indicated that the femur was both larger and weaker 16 weeks after surgery in the ovariectomised group. Results also suggest that the rabbit might be a useful animal model for investigation of diseases related to oestrogen loss such as human postmenopausal osteoporosis. However, additional studies with advanced techniques at several time points via in vivo animal studies, and precision and predictability analyses should be designed to standardise the rabbit as a model for osteoporosis. PMID- 19997667 TI - Long-term follow-up after arthroscopic tenotomy for partial rupture of the biceps brachii tendon. AB - OBJECTIVE: To report the long-term clinical outcomes and radiographic results in dogs diagnosed with partial bicipital rupture and treated by arthroscopic tenotomy. MATERIALS AND METHODS: The medical records of dogs that had undergone arthroscopic tenotomy were retrospectively reviewed. Inclusion criteria for this study were: performance of an arthroscopic tenotomy between August 1999 and July 2007, availability of arthroscopic records data for review, and ability to obtain follow-up data for more than one year after arthroscopic tenotomy. In all cases, owners were interviewed during follow-up appointments or via telephone to determine perceived outcome after surgery. RESULTS: Forty-seven arthroscopic tenotomies were performed on 40 dogs without any major surgical complications. Long-term follow-up examinations, ranging from 12 months to 48 months (mean 26 months) after the tenotomy, were obtained for 24 dogs (25 shoulders). Clinical outcome was assessed as excellent in 22 shoulders, with each dog showing a full return of limb function. A total of 10 dogs (11 joints) were evaluated radiographically; six joints revealed no progression of pathology, and five joints showed a limited progression of pathology. CONCLUSION: Arthroscopic tenotomy in the treatment of bicipital partial rupture yields favourable long term clinical results and a high degree of owner satisfaction. The feasibility of this technique and the long-term clinical and radiographic outcome from our study indicate that this technique can be considered a reliable and safe treatment for partial bicipital rupture. PMID- 19997668 TI - Screw loosening six weeks after triple pelvic osteotomy: comparison of pre-tapped with self-tapping 3.5 mm cortical screws. AB - A retrospective study of 62 dogs that underwent unilateral triple pelvic osteotomy with a Slocum triple pelvic osteotomy plate and Synthes 3.5 mm cortical screws. We compared the probability of screw loosening after six weeks in dogs in which pre-tapped screws were used (30 dogs), with dogs in which self-tapping screws were used (32 dogs). Screw loosening was significantly less prevalent with self-tapping screws than pre-tapped screws. PMID- 19997669 TI - Nutritional secondary hyperparathyroidism in two cats: evaluation of bone mineral density with dual-energy X-ray absorptiometry and computed tomography. AB - Two three-month-old, intact female Abyssinian cats were presented with a history of lameness, constipation and ataxia. The cats had been fed a diet composed almost exclusively of meat. Both showed severe osteopenia and multiple pathological fractures on radiography. Following euthanasia of the more severely affected cat, postmortem examination revealed changes consistent with nutritional secondary hyperparathyroidism and fibrous osteodystrophy, such as cortical thinning, massive connective tissue invasion in the diaphysis of long bones, and hypertrophy of the chief cells in both parathyroid glands. After introducing a balanced commercial diet to the surviving cat, bone mineralisation improved from the baseline value, and at subsequent examinations at three, six and 22 weeks later, as indicated by bone mineral density measurements obtained by dual-energy X-ray absorptiometry and computed tomography. PMID- 19997670 TI - Radiographic features of pelvis and hip joint development of English Bulldogs. AB - OBJECTIVES: To evaluate distinctive features of pelvis and hip joint development of English Bulldogs throughout the first year of life. METHODS: The pelves of 20 English Bulldogs were radiographed at three different ages (<4, 6-8, and 12-14 months). At each time point, the dogs were clinically evaluated and the abnormal hip joints were classified as mild, moderate, or severely deformed. The pelves were compared to a phantom study in which external rotation of a normal hemipelvis around its long axis was artificially created at different degrees, with different pelvic inclinations, and classified as either normal and without deformity, or as mild, moderate, or severely deformed. Hip joints and pelvic scores were statistically compared. RESULTS: Although none of the dogs were considered lame at the end of the study, none of the hips showed normal development; 77.5% were moderately to severely deformed at 12-14 months of age. At this age, 75% of the hemipelves had moderate to severe torsional deformity (>5.2 degrees of external rotation), with retroversion of the acetabulum confirmed by the presence of the crossover sign. An external rotation of the hemipelvis on its long axis >5 degrees was likely associated with a moderate to severely altered hip joint conformation. CLINICAL SIGNIFICANCE: Abnormal hip conformation was common in this series of English Bulldogs. Torsional deformity of the pelves with acetabular retroversion was a common and distinctive feature, which has not yet been thoroughly studied in dogs. These findings need further evaluation in English Bulldogs as well as in other breeds. PMID- 19997671 TI - Correction of a severe torsional malunion of the metacarpus in a calf by transverse osteotomy, transfixation pinning and casting. AB - A four-month-old Holstein heifer was presented for evaluation of an angular limb deviation. Upon initial presentation, the heifer was not clinically lame. An external rotation of 90 degrees of the fetlock was observed in the left forelimb. Radiographs identified a healing closed spiral fracture of the third and fourth metacarpal bones, with a 90 degrees external torsion. A mid-metacarpal bone transverse osteotomy was performed after insertion of transcortical pins in the distal and proximal part of the third and fourth metacarpal bones and in the distal radius. The reduction was performed in order to align both fetlock and carpal joints and a fiberglass cast was applied to complete the external fixation. A satisfactory outcome was associated with the treatment. PMID- 19997672 TI - Ground reaction force profiles after partial and pancarpal arthrodesis in dogs. AB - OBJECTIVES: To evaluate and compare long-term functional outcome after partial carpal arthrodesis and pancarpal arthrodesis in dogs using kinetic gait analysis. METHODS: Fourteen dogs with 19 partial carpal or pancarpal arthrodeses were retrospectively examined and underwent force-plate gait analysis. Mean times since surgery were 29.4 and 24.4 months for pancarpal and partial carpal arthrodesis respectively. Vertical and braking-propulsive ground reaction force profiles were compared between treatment groups, and to those of normal dogs (control group) using Kruskal-Wallis one-way analysis of variance. RESULTS: With the exception of time to vertical peak that occurred earlier in dogs with pancarpal than in dogs with partial carpal arthrodesis (p <0.01), there was no difference between the two treatment groups. Several parameters differed significantly between operated and healthy dogs (p <0.01): vertical impulses were significantly lower in both treatment groups, braking forces and impulses were also reduced after both techniques. Propulsive forces and impulses were only reduced in dogs with pancarpal arthrodesis. When comparing gait parameters of sound limbs of unilateral operated dogs to those of control dogs, braking forces and impulses (p <0.01; p <0.05) were significantly higher in the sound legs of unilateral operated dogs. CLINICAL SIGNIFICANCE: Long-term outcome after partial carpal and pancarpal arthrodesis is good and comparable to each other. Propulsive action may be altered more in dogs with pancarpal arthrodesis. PMID- 19997673 TI - Elbow lameness in dogs of six years and older: arthroscopic and imaging findings of medial coronoid disease in 51 dogs. AB - OBJECTIVES: To report on the frequency and distribution of lesions of the medial coronoid process in dogs of six years of age and older, and to describe the arthroscopic findings in the affected joints. To compare those lesions seen in 'old' dogs with those seen in 'young' dogs between the ages of five and 18 months. METHODS: A retrospective study of dogs six-years-old and older admitted for elbow lameness and subsequent elbow arthroscopy. The dogs were divided into groups according to the lesions of the medial coronoid process diagnosed during arthroscopic examination and computed tomography (CT). The radiographic degree of osteoarthritis (OA) and arthroscopically diagnosed lesions on the medial humeral condyle are described. RESULTS: In 51 'old' dogs, five types of lesions could be identified on arthroscopy and CT: chondromalacia-like lesions (2%), fissures (27.5%), non-displaced fragments (12%), displaced fragments (27.5%), and erosions within the medial compartment without fragmentation (31%). A significantly different distribution of lesions was seen in 'young' dogs: fissures (23%), non displaced fragments (45%), displaced fragments (29%), and erosions within the medial compartment without fragmentation (3%). No difference in radiographic degree of OA was seen between the two groups. CLINICAL SIGNIFICANCE: This study demonstrates the relatively high incidence of medial coronoid disease in dogs older than six years of age, and it highlights one particular problem in 'old' dogs: the complete erosion of the medial compartment. PMID- 19997674 TI - Long-term outcomes of thigh circumference, stifle range-of-motion, and lameness after unilateral tibial plateau levelling osteotomy. AB - Our study evaluated thigh circumference (TC), stifle range of motion (ROM), and lameness in dogs one to five years after unilateral tibial plateau levelling osteotomy (TPLO). We hypothesised that TC, stifle ROM, and lameness would not be different to the unoperated limb (control), one to five years after surgery. Patients that were one to five years post-TPLO were reviewed and were included if they had a unilateral TPLO, and no additional clinical evidence of orthopaedic disease. Standing mid-thigh TC measurements and stifle extension and flexion angles were made in triplicate. Clinical lameness was graded blindly. Data were evaluated statistically using paired t-tests for TC and stifle flexion and extension. Significance was set at p <0.05. Twenty-nine dogs met the inclusion criteria. Mean results for the surgery limbs and control limbs were 39.5 +/- 5.5 cm and 40.1 +/- 5.6 cm for TC, 36.6 +/- 6.8 degrees and 28.6 +/- 4.3 degrees for stifle flexion, and 155.2 +/- 6.6 degrees and 159.8 +/- 4.9 degrees for stifle extension, respectively. The mean TC for the operated limb was 98.5% of the control limb. A significant difference was found between the operated and the control limbs for all measurements. Time after surgery had no apparent affect on outcome. Four of 29 dogs (14%) exhibited some lameness in the TLPO limb during evaluation (one dog was 1 to 2 years postoperative and three dogs were 2 to 3 years postoperative). These results indicate that TC and stifle ROM in the TLPO limb do not return to control-limb measurements one to five years after a TPLO surgery. The clinical significance is unknown as TC returned to 98.5% of control, and the source of lameness in the lame dogs was not identified. PMID- 19997675 TI - Surgical management of large segmental femoral and radial bone defects in a dog: through use of a cylindrical titanium mesh cage and a cancellous bone graft. AB - In this case report, we describe the use of a cylindrical titanium mesh cage combined with cancellous bone graft to surgically manage large segmental bone defects in a dog. A seven-year-old, neutered male cross-breed dog, with highly comminuted fractures of the right femur and the left radius and ulna, was referred for treatment. Previous open reduction and internal fixation of these fractures had failed. Following implant removal and debridement of each bone, a 71 mm segmental femoral defect and a 27 mm segmental radial defect were present. A commercially available cylindrical titanium mesh cage was filled with ss tricalcium phosphate crystals mixed with an equal volume of autogenous cancellous bone graft. The mesh cage was aligned with the proximal and distal parts of each bone using an intramedullary pin passing through the cage, and a locking plate was applied to the proximal and distal fracture fragments to produce compression against the titanium cage. The dog had a successful long-term clinical outcome, and radiographic examination at 22 and 63 weeks after surgery showed the formation of remodelling bridging callus that was continuous across the titanium cage in each of the fractures. Due to the relative simplicity of the technique and the favourable outcome in this case, it should be considered an option when managing comminuted fractures with large bone defects. PMID- 19997676 TI - The effect of the combination of locking screws and non-locking screws on the torsional properties of a locking-plate construct. AB - Little is known about the torsional properties of bone-plate constructs when a combination of locking and non-locking screws have been used. Sixty cadaveric canine femurs were divided into three groups. In the first group, the plate was affixed using three non-locking screws. In the second group, only locking screws were used while a combination of one locking and two non-locking screws were used in the third group. All constructs were subjected to torsion until failure. Torque, angle of torsion, and work were all calculated at the maximum failure point, as well as at five degrees of plastic deformation, which was thought to be more representative of clinical failure. At the maximum failure point, the locking group had significantly higher torque, angle, and work values than the non-locking group. The combination group was intermediate to the two other groups, and significantly differed from the non-locking group in torque, and from the locking group in work. At five degrees of plastic deformation, the locking group required significantly higher torque and work than the non-locking group. The combination group required a significantly higher torque than the non-locking group. This study suggests that a construct composed of all locking screws will fail at a greater torque value, and sustain greater work to failure in torsion compared to a construct composed of all non-locking screws. The addition of a single locking screw to an otherwise non-locking construct will increase the torque at the offset failure point and may be of clinical value in constructs subjected to high torsional loads. PMID- 19997677 TI - Septic polyarthritis caused by Erysipelothrix rhusiopathiae in a dog. AB - A 14-month-old, male German Shepherd dog was admitted with a six-week history of lameness and swelling of the right hindlimb. Clinical examination revealed polyarthritis, fever, petechiae and ecchymoses of the abdominal skin and prepuce. The haematology and blood chemistry were indicative of sepsis. Mediolateral radiographic views of both of the stifle joints revealed signs of bilateral articular capsule swelling. The radiographical, bacteriological and necropsy findings confirmed a diagnosis of septic polyarthritis due to infection with Erysipelothrix rhusiopathiae. PMID- 19997678 TI - [Non recurrent inferior laryngeal nerve: our surgical experience]. AB - AIM: Iatrogenic damage to the recurrent laringeal nerve is the most feared complication in thyroid surgery and the main cause for vocal cord palsy. Prevention of such a lesion is based upon the thorough search for the nerve along its anatomic pathway. In the present study the Authors discuss an important anatomic variation: non-recurrent inferior laryngeal nerve, emphasizing the aspects of surgical anatomy, on the basis of their own experience in neck surgery. MATERIALS AND METHODS: The study takes into conideration 301 surgical interventions on the thyroid gland, in 268 women and 33 men; consisting in 256 total thyroidectomies and 45 loboisthmectomies, as follows: 186 for nodular goiter, 48 for follicular nodule, 34 for papillifer carcinoma, 33 for toxic goiter. The identification and exposure of inferior laryngeal nerve was performed according to the principles and technique by Lahey. RESULTS: The overall prevalence of non recurrent inferior laryngeal nerve was equal to 0.33% (1/301 operations). The anomaly is prevailing on the right side, being due by an anomalous reabsorption of the IV ventral arch. This is the cause of the formation of a subclavian artery that is responsible for dysphagia lusoria. CONCLUSIONS: The rationale for the preservation of recurrent nerve is its systematic identification and exposure along all its course from its laryngeal entry. Diagnosis of non recurrent inferior laryngeal nerve is exclusively intra operative. The anomaly is clinically asymptomatic and its suspicion can be posed in case of vascular anomalies or dysphagia. In such a circumstance the golden diagnostic tool is the angio Tc-RMN, while both the EGDS and baritate esophagous can be of some diagnostic help. PMID- 19997679 TI - [Risk stratification on 206 patients with acute coronary syndrome at Rome Policlinico Umberto I Emergency Department]. AB - OBJECTIVES: The objective of this study on patients with suspected Acute Coronary Disease (ACS) was to verify with the risk stratification any differences between patients in which the AHA/ACC guidelines were not systematically applied and the same patients in which, retrospectively, the AHA/ACC guidelines were adhered to. MATERIALS AND METHODS: Retrospective examination was carried out in our Emergency Department at Rome Policlinico Umberto I on 206 patients (age range 21-88, median age 56.6 +/- 18.9 years) (54.3% males) with symptoms compatible with ACS. All the patients underwent "triage" using code green or code yellow and were sub-divided into 7 subgroups based on degree of risk for death or non-fatal myocardial infarction (MI) at 30 days, (labelled A to G, with A representing highest risk and G the least at-risk) in accordance with the ACC/AHA guidelines. Each patient was then examined for: fi nal outcome, admittance and discharge from hospital, refusal of admittance, death. RESULTS: Of the 206 patients, 48 were judged improperly (23.3%) Of these, 13 were assigned code green rather than code yellow and 11 were assigned code yellow instead of code green. By risk stratification in accordance with the AHA/ACC guidelines, 128 patients (62.1%) belonging to subgroups A,B,C and D should have required hospitalization. Seventy eight patients belonging to subgroups E,F and G should have been sent for observation at the Chest Pain Unit (CPU), 54 from this group would have been potentially discharged after 6-12 hours of negative clinical observation. Actually, hospitalization was requested for 132 patients (64%) of whom 78 accepted, 50 declined and 4 were deceased before admission; 74 patients (35.9%) were discharged after brief observation. Risk stratification for 30 day mortality or non fatal MI emphasizes that 8 of the patients hospitalized should have been treated at the CPU (unwarranted hospitalization). Twenty four of the 50 patients who declined hospitalization should have benefited by brief observation at the Chest pain Unit. Twenty eight of the 78 patients examined in the Emergency Department and later discharged should have been hospitalized (wrong discharge). CONCLUSIONS: This study, even if limited by time restriction, provides enough evidence in support of the effectiveness of the ACC/AHA guidelines to determine subgroups and to correctly determine groups according to level of risk, thus limiting unwarranted hospitalizations and wrong discharges. PMID- 19997680 TI - Percutaneous endoscopic gastrostomy (PEG) in critically ill patients performed at bed in Intensive Care Unit: report of our experience. AB - BACKGROUND AND AIM: Patients with severe brain injures and severe neurological diseases frequently require prolonged nutritional support during their hospitalization as well as during their rehabilitation period. Since 1980, the percutaneous endoscopic gastrostomy (PEG) has become the method of choice for long term feeding. The aim of the present study was to present our experience concerning the placement of PEG in critically ill patients, recovered in Intensive Care Unit (ICU). MATERIALS AND METHODS: From 3-05-2001 to 28-09-2005, 36 patients (13 female, 23 male) with a median age of 63 years [range: 18-86 years], recovered in ICU of the Sandro Pertini Hospital, underwent PEG. These patients were retrospectively evaluated in terms of complications, indications to the procedures, durability of gastrostomy and mortality. Intravenous antibiotic prophylaxis was administered 1 h before the procedure (ceftriaxone 2gr). The entire PEG was placed in ICU at patient's bed, with the assistance of the anaesthetist. Propofol was used e.v. for sedation and fentanest for analgesia while lidocaine was used for local anesthesia. A 16-Fr or 20-Fr tube was inserted by the "pull method", after a complete upper gastroduodenoscopy. RESULTS: PEG was performed mainly for neurological disorders including cerebrovascular accidents (13), SLA (8), post-traumatic coma (7), post-cardiac arrest coma (7) and dementia (1). Procedure related mortality was 0%. The tube was changed in 4 patients due to clogging. The durability of the tube was a median of 2 months (range: 1-12 months). In 23 patients the placement of the PEG was definitive. CONCLUSIONS: Our experience underlines that PEG, in selected critically ill patients, is a safe technique easy to perform even in ICU. PMID- 19997681 TI - [Diabetes worsens spermatic oxidative "stress" associated with the inflammation of male accessory sex glands]. AB - AIMS: Few data in the literature to clarify the true prevalence of sexual male accessory glands inflammations, registered in the diabetic population (DM). Also just-depth are the real pathogenetic mechanisms underlying this association. In a selected population of diabetic patients (DM) with primary infertility and associated clinical condition of sexual male accessory glands infection (MAGI), description of conventional sperm parameters, the concentration of leukocytes and free radicals of oxygen in sperm, the ultrasound andrological characterization, compared to a group of infertile patients with only MAGI. MATERIALS AND METHODS: The study was conducted on 16 patients with type II DM, average age 37 + or - 3 years (27-45 years) with MAGI (WHO criteria, 1993), and primary infertility; 30 patients, average age 33 + or - 4 (21-45 years) with MAGI (WHO criteria, 1993), and primary infertility. RESULTS: The concentration of seminal leukocytes and oxygen free radicals in baseline and after stimulation with specific inducer (f- MLP) in chemiluminescence, in fractions of Percoll 45 and 90% was significantly increased in the group with DM and associated MAGI. The ultrasound evaluation showed impaired emptying of the ampullarvescicular-ductal tract, in the immediate post ejaculatory. CONCLUSIONS: MAGI is a clinically relevant model of spermatic oxidative stress. Our study shows that DM combined with MAGI worsens leukocyte and free oxygen radicals concentration in sperm, without altering the conventional parameters. Also DM seems to favor extension and maintenance of the inflammatory process. PMID- 19997682 TI - Bouveret's syndrome: description of a case. AB - Bouveret's syndrome is a rare condition usually caused by a single large stone impacted in the duodenum. This is a cause of gastric outlet. Even if endoscopy is the mainstay of diagnosis, the radiographic examinations are also important too. Generally, the stones are too large to be removed endoscopically. Conservative endoscopic treatment should be attempted initially, and if it fails, surgical approach should be performed. PMID- 19997683 TI - An interesting case of autonomic dysreflexia. AB - Autonomic dysreflexia (AD) is not an uncommon clinical condition and it is usually detected in patients with complete spinal injuries at or above thoracic 6th vertebral level (T 6). This condition is reported to occur in 48- 60% of cases of spinal cord injury at or above the level of T6. But AD due to injury below T6 is rare. The basic mechanism is thought to be due to excessive, uncontrolled activation of sympathetic system. In the present case, we discuss a persistent AD in 55-yr-old tetraplegic patient with C5 American Spinal Injury Association (ASIA) grade A lesion due to a fall from 10 metre height. MRI examination showed C5 and C6 bi-facets fracture and dislocation with canal compromise. Wiring and fusion was performed but recurrent mucous plugging and aspiration pneumonia and urinary tract infection happened during the hospital stay. Three months later, he was re-admitted with multiple pressure sores, pneumonia, sepsis and high blood pressure. He was administered with nifedepine but the blood pressure kept fluctuating. The present study highlights how the precipitating factors like concomitant urinary tract infection, decubitus ulcers, spasticity triggered the AD attack. The knowledge of the AD and its proper diagnosis and management may be beneficial to all clinicians and the present article attempts to highlight such. PMID- 19997684 TI - In remembrance of Eugene Ladislas Kanabrocki (18 April 1922 * 23 April 2009): a 'General' in the battle of penetrating the normal range and in revealing its relation to the cosmos. AB - Col. Eugene L. (Gene) Kanabrocki, PhD, commanding officer of the 361st Medical Laboratory of the U.S. Army Reserve, together with Col. Lawrence E. (Larry) Scheving, Professor at the University of Arkansas, initiated in May 1969 a linked cross-sectional (hybrid) study at Fort Sam Houston, TX, to examine the oscillatory (circadian) nature of many physiological variables in a group of 13 army men, 22-28 years of age, anticipating that such data would serve, as indeed they did, as time-specified reference values in future investigations of health and aging. In the initial study, 36 variables were examined around the clock in observations at 3-hour intervals. In subsequent 24-hour profiles, mapped in May of 1971 (mostly on new, young subjects, and not officially part of the Aging Project), 1979, 1988, 1993, 1998 and 2003, additional subjects and variables were included. The follow-up studies were conducted at the Hines VA Medical Center in Hines, Illinois. Of the original 13 subjects, four were measured in all 6 studies and another four in 5 of the 6 studies. Three of the eight became diabetic (Type II) and three had vascular circulatory problems. Presently, a bank of circadian data for 187 medically relevant variables of blood (plasma or serum), saliva, urine, vital signs and other variables on the same subjects covers a span of 34 years. Dr. Robert B. Sothern (RBS), of the University of Minnesota, USA, the major analyst of Gene's investigations, in addition to being an add-on subject as he was in three studies, set up the half-hourly monitoring of blood pressure (BP) and heart rate (HR) in the 2003 study that yielded the data suggesting that the standard deviations (SD) of systolic (S) and diastolic (D) BP and HR are infl uenced by a magnetic storm. Since the standard deviation rather than the amplitude of a vascular spectral component was affected, we may be dealing with a stochastic rather than frequency window-dependent resonance with a magnetic storm. Gene and RBS also found (p< 0.08) an about-decadal signature of solar activity in long-acting natriuretic peptide (LANP), vessel dilator (VSDL), insulin, LH, prolactin, T3 uptake and, most importantly, in melatonin (p=0.004), noted solely to constitute a stimulus for follow-up studies, even when resonance occurs in an anticipated Horrebow-Schwabe circadecadal window gauged by relative sunspot (Wolf) numbers and involves many endocrine variables, as anticipated on the basis of independent evidence in melatonin and cortisol. The wealth of circadian information collected in these studies by Gene constitutes a treasure trove of unique advances in the battle of the normal range, with solid contributions also by Prof. Germaine Cornelissen of the University of Minnesota, USA, and by Prof. Ramon C. Hermida of the University of Vigo, Spain. PMID- 19997685 TI - [The chemical neuromodulation of pain: a review]. AB - The chronic pain is a true emergency. In fact, the study "Pain in Europe 2005" showed that 26% of the Italian population is suffering by it. Chronic pain can be benign, when caused by a tissular damage, or malignant when cancer-related. The study of the pain has made a lot of progress in the last years. An example is the chemical neuromodulation, that interferes with the transmission of the pain afferences toward the brain, through the administration of chemical substances in the spinal or cranial compartment in well selected patients. This allows the use of doses lower than those required for other ways of administration, with less collateral effects and a more rapid response. PMID- 19997686 TI - [Bronchoscopy in respiratory intensive care]. AB - Born more than one century ago as a technique for the treatment of the respiratory emergencies due to foreign bodies inhalations, bronchoscopy plays nowadays a crucial and well-defined role in the management of the respiratory critical patient admitted in an intensive hospital setting thanks to its numerous applications both in the diagnostic and in the therapeutic fields. Despite the wide use of the bronchoscopy, especially of fibreoptic technique, it has never to be forgotten the importance of the operator's expertise specifically in this peculiar field of application in order to avoid usefulness risks and to prevent life-threatening consequences for the critical patient. The expert Pulmonologist represents the leader actor for the Clinical Governance of this branch of the Respiratory Intensive Care Medicine. In this review, the Authors examined the aspects connected with the choice of the instrument, the most clinical practice common indications, the risks and the consequences of the use of the bronchoscopy in respiratory intensive care. PMID- 19997687 TI - Effect of pubertal developmental stages and lower limb kinetics during vertical jump task in Sepak Takraw sport. AB - Sepak Takraw is a sport which actively involves the lower limb. A cross sectional study was performed to investigate the lower limb kinetics (ground reaction force, flight time and contact time) involved during the vertical jumping and landing task between the early pubertal and late pubertal stages in National level Sepak Takraw athletes in Malaysia. Twenty athletes (8 pre pubertal and 12 late pubertal) with no previous history of any physical injuries were taken for the study. The kinetics data of the vertical jump task was calculated using a force platform 3-AMTI Biomechanics Force Platform. The Silicon ProCoach (Chart 5 software) was used to collect the ground reaction force signals, flight time and contact time. The results showed a non-significant decrease in peak ground reaction force in the post pubertal group as compared to the pre pubertal group (t(18)=0.659, p=0.518,CI(95)(-2.54, 4.86). Comparison of flight time between the two age groups showed a significant increase in the post pubertal group with mean and SD (0.7773 + or - 0.03) as compared to the pre pubertal group mean and SD (0.7296 + or - 0.05), which was statistically considered significant (p<0.05) (t(18)=-2.401, p<0.05,CI(95)(-0.089, -0.005). The findings from this study demonstrated that the flight time of the vertical jump task differed between the pre and post pubertal athletes with the post pubertal group having an improved ability in maintaining longer duration of flight phase. PMID- 19997688 TI - Children with asthma on inhaled corticosteroids managed in general practice or by hospital paediatricians: is there a difference? AB - AIM: To investigate whether there are differences in asthma characteristics between two populations of children with moderate asthma requiring inhaled corticosteroids (ICS) who are treated in general practice or in hospital practice. PATIENTS AND METHODS: 45 children from general practice and 62 from hospital practice, diagnosed with asthma and treated with ICS, were analysed in terms of lung function parameters, asthma control (ACQ), and use of medication. RESULTS: Children in general practice did not differ significantly from those in paediatric practice with respect to mean age, lung function tests, and corrected daily dose of ICS. The median ACQ score was higher (representing poorer control) in the general practice group than in the paediatric practice group (0.67 and 0.33 respectively, p < 0.05). Fewer children (22.7%) from the general practice group than from the paediatric group (98.4%) had planned review visits (p< 0.01). Prescriptions for a combination ICS/long-acting beta2-agonist (LABA) inhaler were 28.9% in the general practice group and 6.5% in the paediatric group (p<0.05). CONCLUSION: The hospital-based group was better controlled with less frequent use of combination therapy. Our observations stress the necessity for regular review visits for children with moderately severe asthma especially in general practice. PMID- 19997689 TI - Treatment of hidradenitis suppurativa with tumour necrosis factor-alpha inhibitors. AB - Hidradenitis suppurativa (HS) is a common inflammatory skin disease. Medical treatment is often disappointing and in severe disease surgery remains the therapy of choice. Extensive surgery may be effective but also mutilating. Patients experience a significant reduction in quality of life and the need for new treatment modalities are urgent. In recent years patients with HS have been treated off-label with tumour necrosis factor-alpha (TNF-alpha) inhibitors with a varying degrees of effect. We performed a systematic review of papers retrieved from two databases (PubMed and Web of Science) using the follow-ing keywords: hidradenitis suppurativa, acne inversa, infliximab, etanercept, and adalimumab. A total of 34 publications were retrieved, describing treatment of 105 patients. Most cases report treatment with infliximab (52/105). A positive treatment outcome was reported in 90/105 cases, with only 7/105 non-responders and 8/105 patients experiencing side-effects. The side-effects were comparable to those seen in other TNF-alpha inhibitor studies. In the majority of cases the treatment was effective when given as a suppressive therapy, but 15/105 cases were described with long-term remission (>or= 3 months) after the end of therapy. In most publications follow-up was, however, insufficient to allow a systematic exploration of this. TNF-alpha inhibitors seem to be effective in the treatment of HS. However, several questions remain to be answered through specific studies. This review has also identified a need for more standardized reporting of the outcomes as well as randomized controlled trials in this disease. PMID- 19997690 TI - Increased serum tumour necrosis factor-alpha in hidradenitis suppurativa patients: is there a basis for treatment with anti-tumour necrosis factor-alpha agents? AB - Hidradenitis suppurativa (HS) is a recurrent, debilitating suppurative skin disease. Nowadays the major challenge is the choice of optimal treatment. Many conservative therapies seem to have only a supportive character and do not prevent progression of the disease. Early surgical intervention with complete excision of the involved areas is still considered to be the most efficient therapy, but anti-tumour necrosis factor (TNF)-alpha agents may offer a possible non-surgical treatment. The aim of this study was to determine the serum concentration of TNF-alpha and its probable alterations during the disease process in patients with HS. Analysis of TNF-alpha serum concentration in 54 individuals with HS revealed significantly higher levels than in the sera of healthy controls (p = 0.006). To the best of our knowledge, this is the first report of increased TNF-alpha serum concentration in patients with HS. PMID- 19997691 TI - Detection of human herpesvirus 8 in the skin of patients with pityriasis rosea. AB - Pityriasis rosea is an acute self-limiting dermatosis with clinical and epidemiological features that suggest viral involvement. The aim of this study was to investigate a possible association between pityriasis rosea and human herpesvirus 8 (HHV-8). Lesional skin tissue was obtained from 34 Kaposi's sarcoma negative, immunocompetent patients with typical acute phase pityriasis rosea. Nested polymerase chain reaction with specific primer for HHV-8 DNA sequences was performed and all positive results were confirmed by sequencing. Seven out of 34 lesional skin specimens (20.5%) were found to be positive for the HHV-8 genome. All the positive samples were confirmed by DNA sequencing. We conclude that, in some cases, HHV-8 is implicated the pathogenesis of pityriasis rosea. PMID- 19997692 TI - Epidemiology of chronic wound patients and relation to serum levels of mannan binding lectin. AB - The aim of this study was to describe the epidemiology of chronic wounds in a large cohort of patients from a tertiary hospital out-patient clinic, and examine the significance of serum mannan-binding lectin for the occurrence and clinical presentation of such wounds. The study comprised 489 consecutive patients with chronic foot and leg ulcers. A clinical classification of wound- aetiology was performed, and mannan-binding lectin was measured in the sera of patients and healthy controls. The patients presented with 639 wounds altogether; diabetic foot ulcers (309), venous leg ulcers (188), arterial ulcers (109), and vasculitis (33). The mannan-binding lectin levels of patients with venous leg ulcer, alone or in combination with other types of wounds, differed significantly from the control group, and the frequency of values < 100 ng/ml was significantly higher. In diabetic and arterial ulcer patients the frequency of values >or= 3000 ng/ml was significantly higher than that of the control group. This suggests a role for the innate immunity in the pathology of venous leg ulcers, and indicates different roles for mannan-binding lectin in the development of ulcers with different aetiologies; it further suggests that mannan-binding lectin substitution should be tested in a controlled clinical trial. PMID- 19997693 TI - Epidemiological features and costs of herpes zoster in Taiwan: a national study 2000 to 2006. AB - To analyse the epidemiological characteristics and related costs of herpes zoster in Taiwan, a nationally representative cohort of 1,000,000 individuals from the National Health Insurance register was followed up from 2000 to 2006 and their claims data analysed. Overall, 34,280 patients were diagnosed with zoster (incidence 4.89/1000 person-years) and 2944 patients (8.6%) developed post herpetic neuralgia 3 months after the start of the zoster rash (incidence 0.42/1000 person-years). People with older age, diabetes, and immunocompromising conditions were at higher risk of developing zoster and post-herpetic neuralgia. The overall hospitalization rate for zoster was 16.1 cases per 100,000 person years. The cost for each home care case and per hospitalized case were approximately 53.30 euro and 1224.70 euro, respectively. Further research into the cost-effectiveness of zoster vaccine is needed. PMID- 19997694 TI - Drug-induced papuloerythroderma: analysis of T-cell populations and a literature review. AB - Papuloerythroderma of Ofuji is characterized by coalescent solid papules that spare the skin folds. Although cutaneous lymphomas and internal malignancies are known associated conditions, the causative agents are unclear in most cases. A number of recent reports have documente d that drugs can induce papuloerythroderma. We review ed the reported cases and our own cases of drug induc ed papulo erythroderma, together with our data from lympho cyte transformation tests and T-cell subsets of peri pheral blood. All of the 9 patients were male, and the causative drugs were various. Provocation tests were positive in all 6 patients examined. Whereas drug patch tests were negative in all 5 cases tested, the patients' peripheral blood lymphocytes responded well to the culprit drug in 4 of 5 patients tested. The patients had higher percentages of circulating CCR4+CD4+ T helper (Th) 2 cells than CXCR3+CD4+ Th1 cells. Drug induced papuloerythroderma seems to be mediated by Th2 cells reacting with the causative drug. PMID- 19997695 TI - Decreasing prevalence of herpes simplex virus-2 antibodies in selected groups of women in Sweden. AB - High estimates of herpes simplex virus-2 (HSV-2) seroprevalence have been reported for women who were pregnant in Sweden in the 1980s, probably reflecting sexual risk-taking during the 1960s and 1970s. The aim of the present study was to evaluate the HSV-2 seroprevalence in pregnant women and in the female attendees at a clinic for sexually transmitted infections in Sweden at the beginning of the 21st century and to compare the results with those of earlier Swedish studies. Sera were collected during the period 2000 to 2002 from 299 pregnant women at an antenatal clinic and from 290 female attendees at a clinic for sexually transmitted infections in Gothenburg. To enable comparison with earlier seroprevalence data the same test method was used; Helix pomatia antigen in an enzyme-linked immunoassay. The overall HSV-2 prevalence was 10.4% for the pregnant women and 25.2% for the female attendees at the clinic for sexually transmitted infections. The seroprevalence of HSV-2 in pregnant women appears to have decreased in Sweden during the past decade, which may reflect changes in sexual behaviour. PMID- 19997696 TI - Fatal outcome of deep-penetrating lower limb primary cutaneous anaplastic large cell lymphomas in two immunocompromised patients. AB - The occurrence of primary cutaneous anaplastic large cell lymphoma (PCALCL) in immunocompromised patients is rare. Only 11 cases have been reported to date, all of them in organ transplant recipients and none in patient with idiopathic CD4+ T cell lymphocytopaenia. We describe here the original clinical pattern of deep, fascia and muscle-penetrating PCALCL of the lower limb in two immunocompromised patients, one in a renal transplant recipient, the other in a patient with idiopathic CD4+ T-cell lymphocytopaenia. Both patients experienced a negative outcome, contrasting with the usually indolent course of PCALCL in immunocompetent patients, since both died of complications related to the lymphoma 30 and 13 months later, respectively. The unusual clinical aggressiveness of these two cases of PCALCL suggests that, in this peculiar subset with a deep structures involvement hallmark, a worse prognosis could be expected, especially in immunocompromised patients. This information should be taken into consideration when making therapeutic choices. PMID- 19997697 TI - Vincristine, idarubicin, dexamethasone and thalidomide in scleromyxoedema. AB - Scleromyxoedema is a rare disease of unknown aetiology that is characterized by progressive cutaneous mucinosis and paraproteinaemia. A variety of systemic (e.g. gastro intestinal, neurological, pulmonary, cardiac and renal) complications may lead to significant morbidity and mortality necessitating therapeutic intervention. The latter remains challenging. Numerous treatment modalities have been reported in the literature, often, however, with inconsistent responses, frequent relapses and potentially serious side-effects. Moreover, the rarity of scleromyxoedema has prevented the execution of controlled therapeutic trials. This paper discusses current proposed therapeutic strategies and reports the case of a 64-year-old male patient with progressive scleromyxoedema associated with IgG-lambda paraproteinaemia in whom monthly administrations of vincristine, idarubicin and dexamethasone in addition to daily oral thalidomide led to clinical and laboratory remission within 12 weeks. PMID- 19997698 TI - Treatment-resistant Scopulariopsis brevicaulis infection after filler injection. PMID- 19997699 TI - Bowen's disease of the nail matrix presenting as melanonychia: detection of human papillomavirus type 56. PMID- 19997700 TI - Scrofuloderma associated with granuloma annulare-like lichen scrofulosorum. PMID- 19997701 TI - Complementary and alternative medicine: knowledge and attitudes among dermatologists. PMID- 19997702 TI - Eosinophilic cellulitis in a patient with gastric cancer. PMID- 19997703 TI - Cutaneous blastic plasmacytoid dendritic cell neoplasm associated with anaemia and thrombocytopaenia. PMID- 19997704 TI - Syphilitic chancres of the mouth: three cases. PMID- 19997705 TI - Response of ulcerated necrobiosis lipoidica to clofazimine. PMID- 19997706 TI - Cutaneous gamma/delta T-cell lymphoma during treatment with etanercept for rheumatoid arthritis. PMID- 19997707 TI - Purpura fulminans in a patient with rheumatoid arthritis. PMID- 19997708 TI - Subcutaneous phaeohyphomycosis in a renal transplant recipient successfully treated with voriconazole. PMID- 19997709 TI - A finely wrinkled, band-like naevus associated with hyper- and hypopigmented spots: a new entity? PMID- 19997710 TI - HLA typing in an IFN-alpha-induced scar sarcoidosis: possible pathogenetic and clinical implications. PMID- 19997711 TI - Cutaneous Mycobacterium chelonae infection presenting as symmetrical plaques on both shins in an immunocompetent patient. PMID- 19997712 TI - Recurrent angioedema associated with efalizumab. PMID- 19997713 TI - Cauliflower-like lesion on the buttock: a quiz. Pedunculated lipofibroma. PMID- 19997714 TI - Puffy feet in an 11-month-old infant: a quiz. Milroy disease. PMID- 19997715 TI - A crescent-shaped nodule in the temple: a quiz. Nodular fasciitis. PMID- 19997717 TI - [Time intervals during and after emergency room treatment. An analysis using the trauma register of the German Society for Trauma Surgery]. AB - BACKGROUND: The prognosis of severely injured patients depends on a rapid diagnosis and early initiation of therapeutic procedures. MATERIAL AND METHODS: To that end a total of 6,927 prospectively documented severely injured patients with an Injury Severity Score (ISS) > or =16 from the Trauma Registry of the German Trauma Society (DGU, 2002-2007) were analyzed with respect to time intervals during emergency trauma treatment. RESULTS: In cases of indicated emergency surgery the average +/-time in the emergency department was 42+/-34 min, in cases of early surgery 75+/-41 min and in cases of transfer to the intensive care unit (ICU) 83+/-43 min, respectively. The time from the last diagnostic procedure until the end of emergency treatment was 12 min (emergency surgery), 26 min (early surgery) and 32 min (ICU), respectively. Level I (78 min) and level II (72 min) trauma centres showed similar mean times in the emergency department while level III trauma centres had a mean time of 86 min. According to this analysis no general correlation between shorter duration of emergency trauma care and reduced mortality could be observed. CONCLUSION: The duration of time intervals depends on injury severity, treatment after completion of emergency trauma care and the level of the trauma centre. Time management in emergency trauma care can potentially be optimized after completion of the last diagnostic procedure in the emergency room. PMID- 19997718 TI - [Intramedullary nailing of proximal tibial fractures. Complications and risk factors]. AB - INTRODUCTION: We performed a consecutive study on patients with proximal tibial fractures without joint participation to determine the early clinical and radiological outcome. PATIENTS: From January 2000 to December 2005, 22 fractures of the proximal and proximal diaphyseal tibia without joint participation were operated on using the new tibia nail. A total of 18 patients (82%) completed full follow-up after 6 weeks and 3 and 6 months. RESULTS: Two primary and one secondary malalignment were observed Two of them were initially polytraumatised. Furthermore, two patients developed a delayed union, and one non-union occurred. Risk factors for limited bony consolidation were smoking and an open fracture. Screw or nail breakage did not occur, and no infection was observed. PMID- 19997719 TI - Bony Bankart is a positive predictive factor after primary shoulder dislocation. AB - It would be a great advantage if it were possible to categorise the patients with first time dislocations to an initial treatment with the most beneficial outcome. MRI could be a useful method for finding lesions after shoulder dislocation. Fifty-eight patients with traumatic anterior shoulder dislocation were treated by closed reduction and were examined by MRI after a maximum of 2 weeks. The hemarthrosis or effusion present in the joint after the primary dislocation could be used as a contrast for arthrography to identify the lesions present on MRI. At follow-up more than 8 years later, the MRI findings were compared to the shoulder function, shoulder stability, Rowe score and Western Ontario Shoulder Instability Index (WOSI). Besides the age of the patient being above 30, the MRI findings analysed showed that an isolated fracture of the major tubercle, as well as a bony Bankart lesion are prognostic factors for a good functional result and a stable shoulder after a primary dislocation. The glenoid rim fracture was only detected on plain radiographs in 6 out of 10 findings on MRI. MRI findings of a gleniod rim fracture, equal to a bony Bankart lesion, were found to be a prognostic factor for stability and a good functional outcome. PMID- 19997720 TI - A case of intra-articular synovial lipoma of the knee joint causing patellar dislocation. AB - Intra-articular synovial lipoma is rare and clinically manifested mostly by mass effect and occasionally by torsion of the stalk. Patellar dislocation rarely occurs owing to a mass effect of intra-articular tumor of the knee joint. The authors present a case of large intra-articular synovial lipoma at the lateral gutter of the knee joint, which presents as patellar dislocation. A 19-year-old man visited our clinic due to pain and limitation of motion owing to mass at his left knee. MRI revealed an intra-articular soft tissue mass at the lateral gutter of the knee joint with considerable mass effect displacing patella from the trochlear groove. Excisional biopsy confirmed the diagnosis of lipoma, and the final diagnosis was intra-articular synovial lipoma with clinical information. The final radiograph showed well-aligned patella on the trochlear groove. After marginal excision, there was no recurrence or tumor-related morbidity. PMID- 19997722 TI - Effect of blind treatment on stone disease. AB - Most of the drugs administered to stone patients appear to be inappropriate and doing more harm than good to the patients. The objective of this paper is to identify the prevalence of blind chemotherapy among the stone patients and find out the real indication for the drugs administered. Patients who attended the stone clinic for the first time were interviewed to find out what drugs they had been taking before the attendance at the stone clinic. 350 patients consuming specific drugs relevant to stone formation at least for a period of 15 days were selected for a detailed assessment. The type of drug consumed, the dose, the duration, the side effects, compliance rate and effect on stone disease were assessed. The biochemical profile of the patients was assessed to identify the role of the therapeutic modalities utilised. Conclusions regarding the utility of drugs in the process of stone formation were made. The values were compared with those of patients not on medication and considering laboratory standards. Of the 350 patients studied, 96 patients were consuming potassium citrate in different doses, 50 were consuming allopurinol, 44 cystone, 27 potassium citrate + magnesium, 25 calcury, 24 rowatinex, 21 ayurvedic drugs, 17 dystone, 17 homeopathic medicines and 17 other drugs. The longest duration of compliance was for cystone-2.5 years. All other drugs were stopped by the patients themselves due to recurrence of symptoms. As much as 93% of the patients did not feel that there was any significant relief of symptoms. The side effects which prompted the patients to stop medicine were gastro intestinal upset, particularly with potassium citrate, rowatinex and potassium citrate + magnesium combination. The relevant biochemical changes noted were increased urinary citrate levels in patients consuming potassium citrate alone or in combination with magnesium. Serum uric acid was within normal limits in patients consuming allopurinol. Urine uric acid levels were also lower in patients on allopurinol. It is concluded that most of the drugs administered blindly were neither indicated nor beneficial for the patients. Metabolic correction has to be based on proper metabolic assessment. PMID- 19997721 TI - Breathing with the pelvic floor? Correlation of pelvic floor muscle function and expiratory flows in healthy young nulliparous women. AB - INTRODUCTION AND HYPOTHESIS: The aim of this cross-sectional study was to determine correlations between pelvic floor muscle (PFM) function and expiratory function in healthy young nulliparous women. METHODS: In 40 volunteers, PFM function was assessed by vaginal palpation. Forced expiration patterns were evaluated visually and by palpation of the suprapubic insertion region of the anterolateral abdominal muscles. Forced vital capacity (FVC) and forced expiratory flows (FEF) were determined by spirometry. RESULTS: Incremental positive correlation was found between voluntary PFM contraction strength and forced expiratory flow at 25%, 50% and 75% (FEF(25%), FEF(50%), FEF(75%)) of the FVC, respectively. Positive correlation was also found between PFM contraction strength and forced expired volume in 1 s (FEV(1)). No correlation was found between PFM contraction strength and FVC or peak expiratory flow (PEF). CONCLUSIONS: Despite some limitations of this study, the observed correlation between PFM contraction strength and forced expiratory flows may serve as theoretical background for a potential role of coordinated abdominal and PFM training in diseases with expiratory flow limitations. PMID- 19997723 TI - Is high diuresis an important prerequisite for successful SWL-disintegration of ureteral stones? AB - A randomised comparison was made between high and normal diuresis during the primary session of extracorporeal shock wave lithotripsy (SWL) for removal of ureteral stones. High diuresis was accomplished by pressure infusion of approximately 1 L of Ringer acetate solution together with 20 mg of furosemide during the 30-40 min treatment session (Group D). These patients also had a bladder catheter. Patients in Group 0 had a normal diuresis and no bladder catheter. The mean (SD) number of treatment sessions for the 65 patients in Group D were 1.26 (0.51) and for the 60 patients from Group 0 1.35 (0.84) [p > 0.10]. 51 (78%) and 46 (77%) patients in Group D and Group 0, respectively, required only one SWL session. Stones were located in the proximal, mid and distal ureter in 22, 9, and 29 of the patients in Group D and in 22, 5, and 38 in Group 0. No difference in treatment outcome was observed with regard to stone localisation. The stone treatment index (STI(GR)) calculated for Group D and Group 0 were 4.77 and 4.70, respectively. In conclusion there was no obvious advantage of using a forced diuresis during the primary SWL-session for treating patients with ureteral stones. Although a high diuresis and bladder catheter might be beneficial in certain complicated cases, in most situations it seems appropriate to omit this step. PMID- 19997724 TI - Crystal sedimentation and stone formation. AB - Mechanisms of crystal collision being the first step of aggregation (AGN) were analyzed for calcium oxalate monohydrate (COM) directly produced in urine. COM was produced by oxalate titration in urine of seven healthy men, in solutions of urinary macromolecules and in buffered distilled water (control). Crystal formation and sedimentation were followed by a spectrophotometer and analyzed by scanning electron microscopy. Viscosity of urine was measured at 37 degrees C. From results, sedimentation rate (v (S)), particle diffusion (D) and incidences of collision of particles in suspension by sedimentation (I (S)) and by diffusion (I (D)) were calculated. Calculations were related to average volume and urinary transit time of renal collecting ducts (CD) and of renal pelvis. v (S) was in urine 0.026 +/- 0.012, in UMS 0.022 +/- 0.01 and in control 0.091 +/- 0.02 cm min(-1) (mean +/- SD). For urine, a D of 9.53 +/- 0.97 mum within 1 min can be calculated. At maximal crystal concentration, I (S) was only 0.12 and I (D) was 0.48 min(-1) cm(-3) which, even at an unrealistic permanent and maximal crystalluria, would only correspond to less than one crystal collision/week/CD, whereas to the same tubular wall being in horizontal position 1.3 crystals/min and to a renal stone 624 crystals/cm(2) min could drop by sedimentation. Sedimentation to renal tubular or pelvic wall, where crystals can accumulate and meet with a tissue calcification or a stone, is probably essential for stone formation. Since v (S) mainly depends on particle size, reducing urinary supersaturation and crystal growth by dietary oxalate restriction seems to be an important measure to prevent aggregation. PMID- 19997726 TI - Clinical image. Middle interhemispheric variant of holoprosencephaly. PMID- 19997725 TI - Late-onset pulmonary arterial hypertension after a successful atrial or arterial switch procedure for transposition of the great arteries. AB - Common complications after surgery for transposition of the great arteries (TGA) include systemic ventricular dysfunction and arrhythmia after atrial baffle repair (AB) and outflow tract stenosis or regurgitation after the arterial switch (AS). Severe pulmonary hypertension (PHT) is a rarely reported problem after AB and AS. In this study we sought to evaluate the frequency of late onset severe PHT following surgical repair for TGA. We report 3 cases, 2 after AB and 1 after AS, describe the frequency of this complication and treatment response, by comparing the response to pulmonary vasodilators in this group of patients to that of idiopathic or connective tissue disease (CTD) related PHT. We currently follow 85 patients >or=17 years of age with repaired TGA; 77 after AB and 8 after AS. 3.5% of our adult congenital heart disease patients with TGA have developed late severe PHT. None of these patients demonstrated clinical improvement with Bosentan at 6 months, however 2 of 3 were stabilised with the addition of Sildenafil to initial therapy. The third patient died 4 months after the diagnosis of severe PHT, whilst waiting for heart-lung transplantation, despite Bosentan, Sildenafil and inotropic support. By contrast, of 37 patients with idiopathic or CTD related PHT commenced on Bosentan as initial therapy, 32 (86.5%) demonstrated a clinical response at 6 months; the other patients had Sildenafil as added therapy after 6 months. Our data suggest that patients with TGA and late onset PHT are less likely to achieve a clinical response on pulmonary vasodilator monotherapy (P = 0.006). Whilst more investigation is needed, our experience suggests an aggressive clinical course, often requiring combination PHT treatment. PMID- 19997727 TI - Scatter radiation from chest radiographs: is there a risk to infants in a typical NICU? AB - OBJECTIVE: To evaluate the dose of scatter radiation to infants in a NICU in order to determine the minimal safe distance between isolettes. MATERIALS AND METHODS: Dose secondary to scattered radiation from an acrylic phantom exposed to vertical and horizontal beam exposures at 56 kVp was measured at 93 cm and 125 cm from the center of the phantom. This corresponds to 2 and 3 ft between standard isolettes, respectively. For horizontal exposures, the dosimeter was placed directly behind a CR plate and scatter dose at 90-degrees and 135-degrees from the incident beam was also measured. Exposures were obtained at 160 mAs and the results were extrapolated to correspond to 2.5 mAs. Four measurements were taken at each point and averaged. RESULTS: At 125 cm and 93 cm there was minimal scatter compared to daily natural background radiation dose (8.493 microGy). Greatest scatter dose obtained from a horizontal beam exposure at 135 degrees from the incident beam was still far below background radiation. CONCLUSION: Scatter radiation dose from a single exposure as well as cumulative scatter dose from numerous exposures is significantly below natural background radiation. Infants in neighboring isolettes are not at added risk from radiation scatter as long as the isolettes are separated by at least 2 ft. PMID- 19997728 TI - Hirayama disease. PMID- 19997729 TI - Lemierre syndrome. PMID- 19997730 TI - Scan time and patient dose for thoracic imaging in neonates and small children using axial volumetric 320-detector row CT compared to helical 64-, 32-, and 16- detector row CT acquisitions. AB - BACKGROUND: Recently a 320-detector-row CT (MDCT) scanner has become available that allows axial volumetric scanning of a 16-cm-long range (50 cm field of view) in a single 0.35-s rotation. For imaging neonates and small children, volume scanning is potentially of great advantage as the entire scan range can be acquired in 0.35 s, which can reduce motion artefacts and may reduce the need for sedation in clinical CT imaging. Also, because there is no over-ranging associated with axial volumetric scanning, this may reduce patient radiation dose. OBJECTIVE: To evaluate, by means of a phantom study, scan time and patient dose for thoracic imaging in neonates and small children by using axial cone-beam and helical fan-beam MDCT acquisitions. MATERIALS AND METHODS: Paediatric imaging protocols were assessed for a 320-MDCT volumetric scanner (Aquilion ONE, Toshiba, Otawara, Japan). The 320-MDCT scanner allows for cone-beam acquisitions with coverage up to 160 mm, but it also allows for helical fan-beam acquisitions in 64 , 32-, or 16-MDCT modes. The acquisition configurations that were evaluated were 320 x 0.5 mm, 240 x 0.5 mm, and 160 x 0.5 mm for axial volumetric scanning, and 64 x 0.5 mm, 32 x 0.5 mm, and 16 x 0.5 mm for helical scanning. Dose assessment was performed for clinically relevant paediatric angiographic or chest/mediastinum acquisition protocols with tube voltages of 80 or 100 kVp and tube currents between 40 and 80 mA. RESULTS: Scan time was 0.35 s for 320-MDCT acquisitions, scan times varied between 1.9 s and 8.3 s for helical acquisitions. Dose savings varying between 18% and 40% were achieved with axial volumetric scanning as compared to helical scanning (for 320- versus 64-MDCT at 160 mm and 80 kVp, and for 320- versus 16-MDCT at 80 mm and 100 kVp, respectively). Statistically significant reduction in radiation dose was found for axial 320 MDCT volumetric scanning compared to helical 64-, 32-, and 16-MDCT scanning. CONCLUSION: Axial thoracic CT of neonates and small children with volumetric 320 MDCT can be performed between 5 and 24 times faster compared to helical scanning and can save patient dose. PMID- 19997732 TI - Reconstruction of the medial patellofemoral ligament using a longitudinal patellar tunnel technique. AB - Patellar instability is a common clinical problem affecting a young, active population. A large number of procedures have been described to treat patellar instability. We present the clinical results in a case series of 25 medial patellofemoral ligament reconstructions in 21 patients with up to 30 months follow-up (mean: 7.3). Reconstruction was performed using either the gracilis or semitendinosus tendon autograft. The Tegner activity score improved overall from 3 to 4.4 at follow-up and the mean follow-up Kujala score was 87 (range: 55-100). No patella redislocations were observed. Five patients (20%) required a manipulation under anaesthetic but subsequently regained a satisfactory range of motion. Medial patellofemoral reconstruction with both gracilis and semitendinosus tendon graft using a longitudinal tunnel technique provided good post-operative stability restoring the primary soft tissue restraint to pathological lateral patellar displacement with no complications of post operative patellar fracture. PMID- 19997733 TI - Bone allograft provides bone healing in the medial opening high tibial osteotomy. AB - Various materials are used to fill osteotomy defects created in the medial opening wedge high tibial osteotomy (MOWHTO). Our hypothesis was that a bone allograft would provide the osteotomy site bone healing within the expected time. We performed the MOWHTO using a cancellous bone allograft in 310 knees in 284 patients between 2000 and 2005. Internal fixation was achieved with a T-profile AO plate. Preoperative and postoperative radiographic measurements were taken and statistically processed. All patients were followed up for a period ranging from three to eight years, or 5.9 years on average. Implanted cancellous bone allografts 10-fold) in I(SOC) emerged when both STIM1 and Orai1 were co-transfected into HL-7702 cells. Co-overexpression of STIM1 + TRPC1 also caused >10-fold increase in I(SOC), and addition of Orai1 did not cause any further increase. In HL-7702 cells, TRPC1 and Orai1 take part in SOCE independently of each other. Functional interactions of STIM1 and Orai1 or TRPC1 contribute to I(SOC) activation. PMID- 19997764 TI - An NMR structural study of nickel-substituted rubredoxin. AB - The Ni(II) and Zn(II) derivatives of Desulfovibrio vulgaris rubredoxin (DvRd) have been studied by NMR spectroscopy to probe the structure at the metal centre. The beta CH(2) proton pairs from the cysteines that bind the Ni(II) atom have been identified using 1D nuclear Overhauser enhancement (NOE) difference spectra and sequence specifically assigned via NOE correlations to neighbouring protons and by comparison with the published X-ray crystal structure of a Ni(II) derivative of Clostridium pasteurianum rubredoxin. The solution structures of DvRd(Zn) and DvRd(Ni) have been determined and the paramagnetic form refined using pseudocontact shifts. The determination of the magnetic susceptibility anisotropy tensor allowed the contact and pseudocontact contributions to the observed chemical shifts to be obtained. Analysis of the pseudocontact and contact chemical shifts of the cysteine H beta protons and backbone protons close to the metal centre allowed conclusions to be drawn as to the geometry and hydrogen-bonding pattern at the metal binding site. The importance of NH-S hydrogen bonds at the metal centre for the delocalization of electron spin density is confirmed for rubredoxins and can be extrapolated to metal centres in Cu proteins: amicyanin, plastocyanin, stellacyanin, azurin and pseudoazurin. PMID- 19997766 TI - Foot pain in rheumatoid arthritis prevalence, risk factors and management: an epidemiological study. AB - Foot involvement is a major feature of rheumatoid arthritis (RA). Most epidemiological studies of the RA foot report radiological changes and results of clinical examination. This study aimed to determine the prevalence of foot symptoms, frequency of foot assessment and access to foot care from the perspective of people with RA. A questionnaire was sent to 1,040 people with RA throughout the UK enquiring about foot symptoms, their anatomical distribution (via validated mannequins) availability of podiatry services and perceived usefulness of interventions for alleviating foot symptoms. Altogether 585 useable replies were received; 93.5% of respondents reported having experienced foot pain, and 35.4% reported current foot pain as the presenting symptom. Most (68.2%) reported moderate or severe foot pain daily. Pain was most prevalent in the forefoot and/or ankle. The main predictive factors for reporting current foot pain were longer disease duration (mean 13 vs 10.3 years, p = 0.009), higher BMI (25.6 vs 24.1 p = 0.001) and the prevalent foot symptoms foot stiffness and numbness (both p < 0.0001). Age, gender and current treatment were not significantly associated. Most (82%) had discussed foot symptoms with their rheumatologist, and only 64% had seen a podiatrist. Reported current adherence to prescribed orthoses was 55.8% and to prescribed shoes was 29.5%. Foot symptoms are ubiquitous in RA and frequently severe. Most patients had discussed their symptoms with their rheumatologist, and only 64% had specifically seen a podiatrist. Despite the remarkable progress in development of new treatment modalities for RA, foot pain remains a common and disabling symptom. Our findings support the need for wider access to specific foot care services and evidence based, patient-centred interventions. PMID- 19997765 TI - Traumatic displacement of maxillary permanent incisor into the nasal cavity. AB - PURPOSE: The purpose of this paper was to describe a case of unviable alveolar repositioning of an intruded tooth into the nasal cavity and to bring the subject of intrusive tooth injury among patients with dentoalveolar fractures to the attention of trauma surgeons. PATIENT: A 26-year-old male was involved in a car accident and crashed his mouth against the dashboard due to sudden deceleration. Intraoral examination revealed an anterior maxillary dentoalveolar fracture and absence of the central maxillary incisors, right lateral maxillary incisor, and left maxillary canine. Computed tomography showed a dislocated tooth in the nasal cavity. The "missing" left maxillary canine was easily recovered from the floor of the left nostril. CONCLUSIONS: Because complete dislocation of a tooth can cause a frontal sinus abscess, an airway complication, a respiratory tract obstruction, and a complicated lung abscess or sinusitis, anytime a tooth is not accounted for after a dentoalveolar trauma, the possibility that it has been fully intruded should be considered. Computed tomographic scan should be a routine diagnostic study in all cases with associated missing anatomical structures in the oral and maxillofacial region. The need to involve the dental professional in the initial assessment of dental trauma in emergency rooms in hospitals is important in order to identify how many teeth might be missing after dental trauma and to correctly reposition the avulsed teeth when possible. PMID- 19997767 TI - Ultrashort pulsed laser ablation and stripping of freeze-dried dermis. AB - Plasma-mediated laser ablation and dissection of freeze-dried human dermis using an ultrashort pulsed laser of pulse width 900 fs and wavelength 1,552 nm were investigated. The surface ablation line width and depth in relation to irradiation fluence and pulse overlap rate were characterized and measured by scanning electron microscopy. The ablation threshold fluence for freeze-dried dermis was determined as 8.32 J/cm(2) and the incubation factor subject to pulse train irradiation was found to be 0.54. Histological examination showed no thermal damage with single line ablation. Even with multiline ablation, thermal damage was insignificant and the lateral damage zone was generally within 10 microm with 100 continuously repeated line scans. Ultrashort pulsed laser ablation of the interior of dry dermal tissue was shown to strip thin dermal slices with different thicknesses ranging from 20 to 40 microm. PMID- 19997768 TI - Quantitative determination of gymnodimine-A by high performance liquid chromatography in contaminated clams from Tunisia coastline. AB - Quantitative determination by high performance liquid chromatography (HPLC) was performed for gymnodimine-A (GYM-A), a phycotoxin responsible for the contamination of Tunisian clams. This study demonstrates a rapid and reproducible HPLC-ultraviolet (UV) method for extraction, detection and quantification of GYM A in toxic clams. The extraction of GYM-A from the digestive gland of clams in acetone, subsequent clean-up with diethyl ether and extraction with dichloromethane is the more valid protocol. Chromatography analyses were performed using a gradient of acetonitrile-water (10:90 to 90:10), containing trifluoroacetic acid (0.1%) for 20 min at 1 mL/min rate with a C18 column. Recovery rates exceeded 96%, and limits of detection and quantification were 5 ng/mL and 8 ng/g digestive gland, respectively. Repeatability and reproducibility were tested for various samples containing different levels of GYM-A. A significant correlation was observed between toxicity level of samples and the determined amount of GYM-A. Also, the persistence of GYM-A in contaminated clams from Boughrara lagoon was demonstrated. The kinetics discharge study of GYM-A in controlled medium, during 1 month, showed that the process of depuration was biphasic with an exponential discharge of 75% of the total amount of sequestered GYM-A during the first 12 days followed by a slow discharge (>10%) for the subsequent days up to the seventeenth day. This is the first time that a quantitative study of GYM-A in clams from Tunisian coasts is performed through the development of a new method for detection and quantify of this phycotoxin. We found HPLC-UV a reliable and suitable alternative to the mouse bioassay. PMID- 19997770 TI - Effects of deferasirox and deferiprone on cellular iron load in the human hepatoma cell line HepaRG. AB - Two oral chelators, CP20 (deferiprone) and ICL670 (deferasirox), have been synthesized for the purpose of treating iron overload diseases, especially thalassemias. Given their antiproliferative effects resulting from the essential role played by iron in cell processes, such compounds might also be useful as anticancer agents. In the present study, we tested the impact of these two iron chelators on iron metabolism, in the HepaRG cell line which allowed us to study proliferating and differentiated hepatocytes. ICL670 uptake was greater than the CP20 uptake. The iron depletion induced by ICL670 in differentiated cells increased soluble transferrin receptor expression, decreased intracellular ferritin expression, inhibited (55)Fe (III) uptake, and reduced the hepatocyte concentration of the labile iron pool. In contrast, CP20 induced an unexpected slight increase in intracellular ferritin, which was amplified by iron-treated chelator exposure. CP20 also promoted Fe(III) uptake in differentiated HepaRG cells, thus leading to an increase of both the labile pool and storage forms of iron evaluated by calcein fluorescence and Perls staining, respectively. In acellular conditions, compared to CP20, iron removing ability from the calcein Fe(III) complex was 40 times higher for ICL670. On the whole, biological responses of HepaRG cells to ICL670 treatment were characteristic of expected iron depletion. In contrast, the effects of CP20 suggest the potential involvement of this compound in the iron uptake from the external medium into the hepatocytes from the HepaRG cell line, therefore acting like a siderophore in this cell model. PMID- 19997771 TI - Strategies for molecular expression profiling in bladder cancer. AB - Carcinoma of the urinary bladder involves alterations in multiple cellular pathways that dictate the pathology of the disease and clinical outcome of the patient. This includes alterations in regulation of the cell cycle, apoptotic mechanisms, signal transduction and tumor angiogenesis. Interrogation of alterations in multiple molecules associated with these pathways is leading to the development of biomarker panels that are capable of predicting an individual patient's outcome or response to specific treatments. With respect to gene expression profiling, two broad approaches may be identified: a global approach and a pathway-specific approach. The global approach involves a high-throughput effort to profile the entire genome, while the pathway-specific approach quantifies select genes across several pathways. While the former has a high potential for discovery of novel signatures, the latter is important in generating reproducible and concise panels that have the potential for rapid clinical implementation. A combination of both these approaches is needed for the identification and validation of robust marker panels of potential clinical importance in bladder cancer. PMID- 19997769 TI - Identification of an emerin-beta-catenin complex in the heart important for intercalated disc architecture and beta-catenin localisation. AB - How mutations in the protein emerin lead to the cardiomyopathy associated with X linked Emery-Dreifuss muscular dystrophy (X-EDMD) is unclear. We identified emerin at the adherens junction of the intercalated disc, where it co-localised with the catenin family of proteins. Emerin bound to wild type beta-catenin both in vivo and in vitro. Mutating the GSK3beta phosphorylation sites on beta-catenin abolished this binding. Wild type but not mutant forms of emerin associated with X-EDMD were able to reduce beta-catenin protein levels. Cardiomyocytes from emerin-null mice hearts exhibited erroneous beta-catenin distribution and intercalated disc architecture. Treatment of wild type cardiomyocytes with phenylephrine, which inactivates GSK3beta, redistributed emerin and beta-catenin. Emerin was identified as a direct target of GSK3beta activity since exogenous expression of GSK3beta reduced emerin levels at the nuclear envelope. We propose that perturbation to or total loss of the emerin-beta-catenin complex compromises both intercalated disc function and beta-catenin signalling in cardiomyocytes. PMID- 19997773 TI - Aspirin inhibits fractalkine expression in atherosclerotic plaques and reduces atherosclerosis in ApoE gene knockout mice. AB - OBJECTIVE: To determine the fractalkine expression in the aorta of ApoE (-/-) mice and the effect of high-dose aspirin intervention on fractalkine expression and atherosclerotic lesion formation. METHODS: Twenty-one male ApoE gene knockout mice were randomized into three groups to receive either placebo in addition to normal mice chow (n = 7), placebo in addition to a high-fat diet (n = 7), or aspirin (58 mg/kg/d) in addition to a high-fat diet (n = 7). After 12 weeks of study, the mice were euthanized and serum cholesterol, thromboxane B(2), and 6 keto-PGF(1alpha) were examined. Fractalkine and cyclooxygenase expression in aorta were measured and the atherosclerotic lesion analyzed. RESULTS: Pathology image analysis showed that the atherosclerotic plaque was the most extensive in the high-fat diet group while the addition of aspirin greatly reduced the severity of the plaque. Both RT-PCR analysis and immunohistochemical analysis showed that fractalkine expression was the strongest in the high-fat diet group and was significantly decreased by aspirin treatment. Serum thromboxane B(2) was lowered by aspirin while 6-keto-PGF(1alpha) and cholesterol remained unchanged. CONCLUSIONS: The results of our study indicate that high dose aspirin can improve the atherosclerotic lesion and suppress the fractalkine expression in murine aorta. PMID- 19997774 TI - The DSM diagnostic criteria for sexual sadism. AB - I reviewed the empirical literature for 1900-2008 on the paraphilia of Sexual Sadism for the Sexual and Gender Identity Disorders Workgroup for the forthcoming fifth edition of the Diagnostic and Statistical Manual of Mental Disorders (DSM). The results of this review were tabulated into a general summary of the criticisms relevant to the DSM diagnosis of Sexual Sadism, the assessment of Sexual Sadism utilizing the DSM in samples drawn from forensic populations, and the assessment of Sexual Sadism using the DSM in non-forensic populations. I conclude that the diagnosis of Sexual Sadism should be retained, that minimal modifications of the wording of this diagnosis are warranted, and that there is a need for the development of dimensional and structured diagnostic instruments. PMID- 19997772 TI - Diverse roles of the actin cytoskeleton in striated muscle. AB - In addition to the highly specialized contractile apparatus, it is becoming increasingly clear that there is an extensive actin cytoskeleton which underpins a wide range of functions in striated muscle. Isoforms of cytoskeletal actin and actin-associated proteins (non-muscle myosins, cytoskeletal tropomyosins, and cytoskeletal alpha-actinins) have been detected in a number of regions of striated muscle: the sub-sarcolemmal costamere, the Z-disc and the T tubule/sarcoplasmic reticulum membranes. As the only known function of these proteins is through association with actin filaments, their presence in striated muscles indicates that there are spatially and functionally distinct cytoskeletal actin filament systems in these tissues. These filaments are likely to have important roles in mechanical support, ion channel function, myofibrillogenenous and vesicle trafficking. PMID- 19997775 TI - Effect of autogenic training on general improvement in patients with irritable bowel syndrome: a randomized controlled trial. AB - Autogenic training (AT) is a useful and comprehensive relaxation technique. However, no studies have investigated the effects of AT on irritable bowel syndrome (IBS). In this study we tested the hypothesis that AT improves symptoms of IBS. Twenty-one patients with IBS were randomly assigned to AT (n = 11, 5 male, 6 female) or control therapy (n = 10, 5 male, 5 female). AT patients were trained intensively, while the control therapy consisted of discussions about patients' meal habits and life styles. All patients answered a question related to adequate relief (AR) of IBS symptoms and four questionnaires: Self-induced IBS Questionnaire (SIBSQ), Self-reported Depression Scale (SDS), State-Trait Anxiety Inventory (STAI), and Medical Outcome Short Form 36 Health Survey (SF-36). The proportion of AR in the last AT session in the AT group (9/11, 81.8%) was significantly higher than that in the controls (3/10, 30.0%, Chi-square test, p = 0.048). Two subscales of the SF-36, i.e., social functioning and bodily pain, were significantly improved in the AT group (p < 0.05) as compared to the control group. Role emotional (p = 0.051) and general health (p = 0.068) showed a tendency for improvement in the AT group. AT may be useful in the treatment of IBS by enhancing self-control. PMID- 19997777 TI - Changes in endogenous estrogens and expression of behaviors associated with estrus during the periovulatory period in Heatsynch treated Murrah buffaloes (Bubalus bubalis). AB - The objective of this study was (1) to establish the duration of behavioral estrus signs and timing of ovulation in Murrah buffaloes (n = 10) and (2) to determine relationship between behavioral estrus signs with change in plasma estrogen concentrations in animals treated with Heatsynch protocol. Estrus and its behavioral signs were detected at hourly intervals by visual observations per rectal examination of genitalia and bull parading four times in a day for 30 min each. Among the behavioral signs of estrus, swollen vulva was the best indicator of estrus followed by excitement and chasing by bull (90%). Among the duration of behavioral estrus signs, the first and longest duration of estrus signs was swollen vulva, which was seen up to 21.6 +/- 1.1 h after onset of estrus. The mean total duration of estrus symptoms from appearance to disappearance of all the behavioral estrus symptoms was 40.2 +/- 1.8 h. Endocrine profile during the periestrus period showed that the mean peak concentrations of total estrogen 330.9 +/- 108.3 pg/ml occurred at 9.6 +/- 1.0 h after estradiol benzoate injection. The average number of estrus symptoms observed per animal during Heatsynch treatment was 7.1. Ovulation occurred after 50.0 +/- 2.0 h after estradiol benzoate treatment and 26.7 +/- 2.0 h after end of total estrogen surge, respectively. In conclusion, our results suggest that all signs of behavioral estrus occurred after the preovulatory rise in estrogens. The first sign of estrus was swollen vulva, and this symptom persisted the longest. PMID- 19997778 TI - Can medicalization be good? Situating medicalization within bioethics. AB - Medicalization has been a process articulated primarily by social scientists, historians, and cultural critics. Comparatively little is written about the role of bioethics in appraising medicalization as a social process. The authors consider what medicalization means, its definition, functions, and criteria for assessment. A series of brief case sketches illustrate how bioethics can contribute to the analysis and public policy discussion of medicalization. PMID- 19997776 TI - Brain arachidonic acid cascade enzymes are upregulated in a rat model of unilateral Parkinson disease. AB - Arachidonic acid (AA) signaling is upregulated in the caudate-putamen and frontal cortex of unilaterally 6-hydroxydopamine (6-OHDA) lesioned rats, a model for asymmetrical Parkinson disease. AA signaling can be coupled to D(2)-like receptor initiated AA hydrolysis from phospholipids by cytosolic phospholipase A(2) (cPLA(2)) and subsequent metabolism by cyclooxygenase (COX)-2. In unilaterally 6 OHDA- and sham-lesioned rats, we measured brain expression of cPLA(2), other PLA(2) enzymes, and COX-2. Activity and protein levels of cPLA(2) were significantly higher as was COX-2-protein in caudate-putamen, frontal cortex and remaining brain on the lesioned compared to intact side of the 6-OHDA lesioned rats, and compared to sham brain. Secretory sPLA(2) and Ca(2+)-independent iPLA(2) expression did not differ between sides or groups. Thus, the tonically increased ipsilateral AA signal in the lesioned rat corresponds to upregulated cPLA(2) and COX-2 expression within the AA metabolic cascade, which may contribute to symptoms and pathology in Parkinson disease. PMID- 19997779 TI - Soft palate length and upper airway relationship in OSA and non-OSA subjects. AB - BACKGROUND: The narrowest area of the airway between the posterior nasal opening and the epiglottis is usually located in the retro palatal area. Many consider this the most likely site of airway obstruction during an obstructive sleep apnea (OSA) event. The aim of this study was to investigate the differences in soft palate and airway length between OSA and non-OSA patients. METHODS: In this study, we analyzed the ratio of the soft palate and the upper airway length in 45 consecutive patients. Twenty-five had an Apnea-Hypoapnea Index of more than five events per hour and were classified in the OSA group (male, 19; female, 6). These patients were compared with 20 normal controls (male, 12; female, 8). Controls who complained of snoring did have sleep studies (n=5). The other fifteen controls were clinically asymptomatic and did not have sleep studies. Medical computed tomography scans were taken to determine the length of the upper airway and the soft palate length measured in the midsagittal image. RESULTS: Soft palate length was significantly larger in OSA patients compared to controls (p=0.009), and in men compared to women (p=0.002). However, there were no differences in airway length. The soft palate length, as a percent of oropharyngeal airway length, was significantly larger in OSA patients compared to controls (p= <0.0001) and in men compared to women (p=0.02). Soft palate length increases significantly with age by 0.3 mm per year in males (after adjustment for body mass index (BMI) and OSA). Soft palate length as a percent of airway length is larger in OSA patients and increases significantly with BMI in males only after adjusting for age. CONCLUSION: In this study, OSA patients had a longer soft palate in proportion to their oropharyngeal airway compared to controls as well as men compared to women. This proportion could be used for identifying patients at risk for OSA in combination with age. PMID- 19997782 TI - Sutures. PMID- 19997780 TI - Sex ratio deviation of offspring in the area where exposure to the processing of electronic wastes. PMID- 19997781 TI - Several dystrophin-glycoprotein complex members are present in crude surface membranes but they are sodium dodecyl sulphate invisible in KCl-washed microsomes from mdx mouse muscle. AB - The dystrophin-glycoprotein complex (DGC) is a large trans-sarcolemmal complex that provides a linkage between the subsarcolemmal cytoskeleton and the extracellular matrix. In skeletal muscle, it consists of the dystroglycan, sarcoglycan and cytoplasmic complexes, with dystrophin forming the core protein. The DGC has been described as being absent or greatly reduced in dystrophin deficient muscles, and this lack is considered to be involved in the dystrophic phenotype. Such a decrease in the DGC content was observed in dystrophin deficient muscle from humans with muscular dystrophy and in mice with X-linked muscular dystrophy (mdx mice). These deficits were observed in total muscle homogenates and in partially membrane-purified muscle fractions, the so-called KCl-washed microsomes. Here, we report that most of the proteins of the DGC are actually present at normal levels in the mdx mouse muscle plasma membrane. The proteins are detected in dystrophic animal muscles when the immunoblot assay is performed with crude surface membrane fractions instead of the usually employed KCl-washed microsomes. We propose that these proteins form SDS-insoluble membrane complexes when dystrophin is absent. PMID- 19997783 TI - Barrett's esophagus: a late complication of laparoscopic adjustable gastric banding. AB - Laparoscopic adjustable gastric banding has become a popular bariatric restrictive procedure in the USA. The increasing popularity of the laparoscopic adjustable gastric band procedure could, in part, be related to the lower cost and lower morbidity compared with laparoscopic gastric bypass. Although its placement is related to a lower number of perioperative complications compared with laparoscopic gastric bypass, its morbidity may be substantial. Barrett's esophagus or esophageal intestinal metaplasia is a known complication of chronic gastro-esophageal reflux disease that, in rare occasions, progresses to dysplasia and esophageal adenocarcinoma. Barrett's esophagus, after laparoscopic adjustable gastric banding placement, is a rare but not unexpected complication after gastric band placement. The incidence of Barrett's esophagus after adjustable gastric banding is not known. We present a case of Barrett's esophagus as a result of laparoscopic adjustable gastric banding placement due to a chronically and highly restrictive gastric band in a former morbidly obese patient. PMID- 19997784 TI - Increased serum nitric oxide concentration after bariatric surgery--a potential mechanism for cardiovascular benefit. AB - BACKGROUND: It is believed that endothelial dysfunction associated with obesity contributes to reduced vascular production of nitric oxide (NO). Weight reduction after bariatric surgery is known to decrease the risk of cardiovascular disease. The purpose of this study was to determine whether bariatric surgery leads to improvement of metabolic markers of endothelial function: serum NO and its precursor (arginine) concentrations in obese patients. METHODS: Serum NO and L arginine concentrations were measured in 25 morbidly obese patients directly before and 6 months after bariatric surgery. Moreover, selected parameters that may be involved in development of endothelial dysfunction were also studied. Control group consisted of ten healthy individuals with normal body weight. RESULTS: Six months after bariatric surgery, serum NO concentration was approximately 40% higher than before surgery. Surprisingly, serum NO concentration in nonobese controls was essentially similar to obese patients before surgery. In contrast, serum L-arginine concentration was higher in obese patients than in controls and decreased significantly after surgery. The body weight, blood pressure, triacylglycerols, LDL/HDL-cholesterol ratio, insulin, homeostasis model assessment score (HOMA-index), C-reactive protein, and white blood cell count were higher in obese patients as compared with controls and decreased significantly after surgery. CONCLUSIONS: Our results indicate that improvement of insulin resistance, lipidemia, and blood pressure as well as reduction of systemic inflammation after bariatric surgery were associated with the increase of serum NO concentration. We propose that the increase in serum NO concentration contribute to diverse beneficial effects of weight loss after bariatric surgery especially in the context of risk of atherosclerosis. PMID- 19997785 TI - Pilot data on the endoscopic placement of covered metal stents to treat gastrojejunal leaks in a porcine model. AB - BACKGROUND: Placement of stents may be an alternative option to treat gastrojejunal (GJ) anastomotic leaks after Roux-en-Y gastric bypass (RYGB) surgery. The aim was to evaluate the performance of a covered metal stent (Alimaxx-E, 18 x 100 mm) across a GJ leak in a porcine model. METHODS: In eight pigs, a RYGB was performed and a 1-cm leak was created. A retrogastric and anterogastric GJ was performed equally with four pigs in each group. Stents were placed across the GJ anastomosis using endoscopy and fluoroscopy. The endpoints were position of the stents, healing of the leak, and complications at necropsy. RESULTS: Stent placement was feasible in all pigs it was attempted. In the retrogastric GJ group, pigs without stents survived 2 weeks without complications. Two pigs with stents died on postoperative days (PODs) 4 and 5. In the anterogastric GJ group, one pig with one stent survived 2 weeks with healing of the leak but with stent migration into the Roux limb. The remaining three pigs died between PODs 4 and 6. Pressure necrosis of the Roux limb caused by the distal end of the stent was found in all. CONCLUSIONS: Placement of fully covered metal stents is feasible. The use of small pigs with a thin small bowel wall in combination with relatively large stents was most likely the reason for complications found in all stented animals. Further studies of GJ leaks using these stents will require larger pigs or a different species. PMID- 19997786 TI - Laparoscopic sleeve gastrectomy as a single-stage bariatric procedure. AB - BACKGROUND: Laparoscopic sleeve gastrectomy is increasingly being used as a stand alone procedure in bariatric surgery, with medium-term follow-up data now emerging. We present our early experience in patients with a mean body mass index (BMI) in the super-obese range. METHODS: Review of prospectively collected data for the first 100 patients who underwent laparoscopic sleeve gastrectomy at Counties Manukau District Health Board between March 2007 and July 2008. RESULTS: One hundred patients were identified, with a mean age of 43 years (range, 20-60 years). Maori and Pacific Islanders made up 31% of the patient subset. Patients had a mean BMI of 50.3 kg/m(2) (range, 34.5-72.8 kg/m(2)). Forty-five patients were super-obese. The median hospital stay was 2 days (range, 1-7 days). Mean follow-up was 12.0 months (range, 0.9-23.3 months). Mean excess weight loss was 62.9% (range, 7.2-129.0%). Twenty-five percent of patients were diabetic and 45% of patients were hypertensive pre-operatively. Diabetics and hypertension resolved or improved in 72% and 60% of patients, respectively. There was a major complication rate of 7%, including three staple-line leaks (one requiring laparotomy), two staple-line bleeds (one requiring laparotomy) and one infected haematoma. There were no deaths. CONCLUSIONS: In this public hospital setting, laparoscopic sleeve gastrectomy has achieved satisfactory weight loss results with an acceptable complication rate in the medium-term. PMID- 19997787 TI - Production of cellulase from kraft paper mill sludge by Trichoderma reesei rut C 30. AB - Paper mill sludge is a solid waste material generated from pulping and papermaking operations. Because of high glucan content and its well-dispersed structure, paper mill sludges are well suited for bioconversion into value-added products. It also has high ash content originated from inorganic additives used in papermaking, which causes hindrance to bioconversion. In this study, paper mill sludges from Kraft process were de-ashed by a centrifugal cleaner and successive treatment by sulfuric acid and sodium hydroxide, and used as a substrate for cellulase production. The treated sludge was the only carbon source for cellulase production, and predominantly inorganic nutrients were used as the nitrogen source for this bioprocess. The cellulase enzyme produced from the de ashed sludge exhibited cellulase activity of 8 filter paper unit (FPU)/mL, close to that obtainable from pure cellulosic substrates. The yield of cellulase enzyme was 307 FPU/g glucan of de-ashed sludge. Specific activity was 8.0 FPU/mg protein. In activity tests conducted against the corn stover and alpha-cellulose, the xylanse activity was found to be higher than that of a commercial cellulase. Relatively high xylan content in the sludge appears to have induced high xylanase production. Simultaneous saccharification and fermentation (SSF) was performed using partially de-ashed sludge as the feedstock for ethanol production using Sacharomyces cerevisiae and the cellulase produced in-house from the sludge. With 6% (w/v) glucan feed, ethanol yield of 72% of theoretical maximum and 24.4 g/L ethanol concentration were achieved. These results were identical to those of the SSF using commercial cellulases. PMID- 19997788 TI - In the shadow of Darwin: Anton de Bary's origin of myxomycetology and a molecular phylogeny of the plasmodial slime molds. AB - In his Origin of Species (John Murray, London, 1859), Charles Darwin described the theory of descent with modification by means of natural selection and postulated that all life may have evolved from one or a few simple kinds of organisms. However, Darwin's concept of evolutionary change is entirely based on observations of populations of animals and plants. He briefly mentioned 'lower algae', but ignored amoebae, bacteria and other micro-organisms. In 1859, Anton de Bary, the founder of mycology and plant pathology, published a seminal paper on the biology and taxonomy of the plasmodial slime molds (myxomycetes). These heterotrophic protists are known primarily as a large composite mass, the plasmodium, in which single nuclei are suspended in a common 'naked' cytoplasm that is surrounded by a plasma membrane. Here we summarize the contents of de Bary's 1859 publication and highlight the significance of this scientific classic with respect to the establishment of the kingdom Protoctista (protists such as amoebae), the development of the protoplasmic theory of the cell, the introduction of the concept of symbiosis and the rejection of the dogma of spontaneous generation. We describe the life cycle of the myxomycetes, present new observations on the myxamoebae and propose a higher-order phylogeny based on elongation factor-1 alpha gene sequences. Our results document the congruence between the morphology-based taxonomy of the myxomycetes and molecular data. In addition, we show that free-living amoebae, common protists in the soil, are among the closest living relatives of the myxomycetes and conclude that de Bary's 'Amoeba-hypothesis' on the evolutionary origin of the plasmodial slime molds may have been correct. PMID- 19997790 TI - Socioeconomic status and psychosocial mechanisms of lifestyle change in a type 2 diabetes prevention trial. AB - BACKGROUND: Little is known about psychosocial mechanisms that may underlie differences in lifestyle change between socioeconomic groups. PURPOSE: The purpose of this study is to examine how educational level influences middle-aged participants' (N = 385) psychosocial responses to the GOAL Lifestyle Implementation Trial. METHODS: The measurements of self-efficacy and planning for healthy lifestyle were conducted pre-intervention (T1) and post-intervention (T2, 3 months), and measurements of exercise and healthy eating as outcomes at T1 and at 12 months (T3). RESULTS: Psychosocial determinants at T1 and their T1-T2 changes were mostly similar, irrespective of educational levels. Exercise barriers self-efficacy was enhanced slightly less (p = 0.08) among the low-SES. T2 levels as well as pre-post-intervention changes in exercise self-efficacy predicted 12-month changes in exercise, and T2 diet coping planning predicted changes in dietary fat intake. The associations were similar across all SES groups. CONCLUSIONS: Enhancing self-efficacy and planning is similarly effective among intervention participants regardless of education level. PMID- 19997789 TI - Challenges in umbilical cord blood stem cell banking for stem cell reviews and reports. AB - Twenty years has passed since the first report of a successful cord blood transplant was reported in 1989 in a child with Fanconi's anemia. During these 20 years, the cord blood field has had dramatic growth, with over 400,000 cord blood units donated and stored worldwide for unrelated use. Approximately, 14,000 unrelated cord blood transplants have been performed to date for patients with hematologic malignancies and bone marrow disorders, and who do not have matched family or unrelated donors. In contrast, about 900,000 cord blood units have been stored privately for personal use, with about 100 autologous transplants performed. Twenty years ago, due to the low cell dose, cord blood transplants were only performed in children. Today, with the use of better banking techniques, reduced intensity transplants, and double cord blood transplantation, the majority of cord blood transplants are being performed in adults. In this chapter, we review the scientific basis for cord blood transplantation, and outcome data in both pediatric and adult transplantation. We will then focus on the recent concerns regarding private and public cord blood banking. Finally, we discuss the future of cord blood transplantation, and the exciting work beginning outside of oncology. PMID- 19997791 TI - [Development of cardiovascular surgery]. AB - On the occasion of the 80th anniversary of the journal "Der Chirurg" (The Surgeon) this article summarizes the development of cardiac surgery. Beginning from the first cardiac suture in 1897 by Ludwig Wilhelm Carl Rehn, through the first catheter investigation of the heart by Werner Forssmann in 1923 to the year 2008 when nearly 100,000 cardiac interventions were carried out in Germany and of these some 90,000 using a heart-lung machine. The article describes the founding of the German Society for Thorax, Heart and Vascular Surgery in the year 1971 and the reintegration in the German Society for Surgery, stemming from efforts for the unification of the structure of further education (common trunk). The motto for this process, which was actively supported by both societies, was "Independence when necessary, cooperation when possible". PMID- 19997792 TI - [Analysis of satisfaction of referrers of patients with rectal carcinoma to surgical clinics]. PMID- 19997794 TI - [Ostomy for stool deviation: indications and operative techniques]. AB - The deviation of feces is a very old and still largely applied technique used by general surgeons. Indications divide into elective and emergency operations. Among the elective cases, rectal carcinoma, extensive perianal fistulae and neurogenic disorders of the distal colon are the most common indications. In an emergency situation causes such as extensive peritonitis (e.g. due to anastomotic leakage) and extensive trauma to the pelvis/rectum quite often result in a stoma. All segments of the intestine, from the first loop after the ligament of Treitz down to the sigmoid colon, are feasible for a deviation procedure depending on the indications and the anatomical conditions. The decision whether to create a loop ostomy or a terminal stoma is reached with respect to the underlying indications and the anatomical and pathophysiological situation. The aim of the procedure is a complete and adequate stool deviation as well as a situation where the patient is able to take care of the ostomy in a convenient and reliable fashion while guided and educated by a stoma therapist. The prevention and treatment of complications, such as stoma necrosis or retraction are absolutely crucial in order to have satisfying long-term results and an acceptable quality of life. PMID- 19997795 TI - Potentials of regenerative medicine for liver disease. AB - Liver transplantation is still the only effective treatment for end-stage liver disease. However, because of the serious worldwide shortage of donated organs, an alternative cellular therapy would be desirable. Animal studies and preclinical trials have indicated that hepatocyte transplantation can serve as an alternative to liver transplantation. Unfortunately, however, the harvesting of hepatocytes is associated with the same problem as organ transplantation, i.e., a lack of a suitable cell source. Therefore, current stem cell technology, which is attempting to establish an unlimited supply of hepatocytes, would facilitate the clinical application of hepatocyte transplantation. This review summarizes current knowledge of embryonic and adult stem cell differentiation into hepatocytes and discusses how liver stem cells could be applied clinically in the future. PMID- 19997796 TI - The possibility of performing a limited resection and a lymphadenectomy for proximal gastric carcinoma based on sentinel node navigation. AB - PURPOSE: This study examined the possibility of performing a limited resection and a lymphadenectomy with sentinel node navigation surgery (SNNS) for the treatment of proximal gastric carcinoma. METHODS: Thirty patients with cT1N0 (n = 23) and cT2N0 (n = 7) proximal gastric carcinoma that was located primarily in the U area (the upper third of the stomach) were enrolled. indocyanine green (ICG; 0.5 ml) was injected endoscopically into the submucosa of the four quadrants encompassing the cancer. Twenty minutes after injection, infrared ray electronic endoscopy (IREE) was used to identify the lymph nodes that were stained with ICG (sentinel nodes, SNs) around the serosa and surrounding fat tissue. RESULTS: One hundred percent of the SNs were identified with our SNNS method. The most common location of SNs was No. 3 (T1: 78%, T2: 100%). The main route of lymphatic drainage was from No. 1 or No. 3 to No. 7 (T1: 95%, T2: 100%). In T1 cancer, Indocyanine green was not distributed to the right gastric area, and no patients had SNs in No. 5 or No. 8a. Four cT2 cancer patients had lymph node metastases, all of which were SNs. There were no cases of postoperative metastasis or recurrence. CONCLUSIONS: For the cT1 proximal gastric carcinoma patients, limited dissection of the ICG tracer-positive lymphatic areas alone by SNNS using IREE may be acceptable. The main lymphatic drainage route of proximal gastric carcinoma is the left gastric artery area (Nos. 1, 3, and No. 7) and dissection of this area is important. PMID- 19997797 TI - Short-term intravenous antimicrobial prophylaxis in combination with preoperative oral antibiotics on surgical site infection and methicillin-resistant Staphylococcus aureus infection in elective colon cancer surgery: results of a prospective randomized trial. AB - PURPOSE: We performed a prospective randomized study to assess the effectiveness of short-term intravenous antimicrobial prophylaxis in combination with preoperative oral antibiotics on a surgical site and methicillin-resistant Staphylococcus aureus (MRSA) infection in elective colon cancer surgery. METHODS: The patients were administered preoperative oral antibiotics, kanamycin and erythromycin, after mechanical cleansing, which began within 24 h of elective surgery for colon cancer. The patients were randomly assigned to receive the intravenous administration of cefmetazol or cefotiam on the day of surgery (group 1) or for 3 days (group 2). A total of 275 patients (136 for group 1 and 139 for group 2) were eligible for the study. RESULTS: The incidence of a surgical site infection was 5.1% in group 1 and 6.5% in group 2 (P = 0.80). The incidence of MRSA infection was 2.2% in group 1 and 2.9% in group 2 (P > 0.99). A multivariate logistic regression analysis showed that the American Society of Anesthesiologists physical status score and the duration of surgery were independent significant factors affecting the surgical site infection and MRSA infection. CONCLUSION: These findings suggest that short-term intravenous antimicrobial prophylaxis in combination with preoperative oral antibiotics may be successfully applied to colon cancer surgery that is generally performed in Japan. PMID- 19997798 TI - Retrospective, matched case-control study comparing the oncologic outcomes between laparoscopic surgery and open surgery in patients with right-sided colon cancer. AB - PURPOSE: The short- and long-term outcomes of laparoscopic surgery for right sided colon cancer remain largely uninvestigated. This study was undertaken to compare the morbidity and mortality after either a laparoscopic right hemicolectomy (LRHC) or an open right hemicolectomy (ORHC) for this type of tumor. METHODS: The study group included 100 patients who underwent an LRHC and 100 patients who underwent an ORHC for right-sided colon cancer from 1990 through 2004. The two groups were retrospectively well matched with respect to sex, age (+/-5 years), and pathological tumor-node-metastasis (TNM) stage. RESULTS: The median follow-up period was 83 months in the LRHC group and 105 months in the ORHC group. The LRHC group had a lower volume of intraoperative bleeding (P < 0.001), a lower rate of wound infection (P = 0.019) or postoperative intestinal obstruction (P = 0.013), and a shorter hospital stay (P < 0.001) than the ORHC group. The rate of recurrence did not differ significantly between the LRHC group (19%) and the ORHC group (22%). In patients with TNM stage I or II, the disease free survival (DFS) rate (94.9% vs 95.1%) and overall survival (OS) rate (95.8% vs 95.0%) did not differ significantly between the two groups. A similar tendency was observed in patients with stage III with the rates for DFS (71.3% vs 60.4%) and OS (73.6% vs 64.1%), respectively. CONCLUSIONS: An LRHC for right-sided colon cancer has the advantage over an ORHC of better short-term outcomes, and both groups have similar long-term oncologic outcomes. An LRHC is thus an acceptable alternative to an ORHC for the treatment of this type cancer. PMID- 19997799 TI - TSU68, an antiangiogenic receptor tyrosine kinase inhibitor, induces tumor vascular normalization in a human cancer xenograft nude mouse model. AB - PURPOSE: Combination therapy using antiangiogenic and cytotoxic agents is a useful strategy for advanced cancer, but the mechanism has not yet been elucidated. Moreover, there is a persistent paradox that destroying tumor vasculature with antiangiogenic agents disturbs the delivery of cytotoxic agents. It has been hypothesized that antiangiogenic agents can lead to normalization of tumor vessels that are structurally and functionally abnormal. The normalization means enhancing the deliver of cytotoxic agents. Our purpose was to investigate whether TSU68, a multiple receptor tyrosine kinase inhibitor that targets vascular endothelial growth factor receptor-2 (VEGFR2), platelet-derived growth factor receptor (PDGFR), and fibroblast growth factor receptor (FGFR), would induce the normalization of tumor vessels. METHODS: TSU68 was administered for 7 days to mice with xenografted tumors. Tumors of interstitial fluid pressure (IFP) were measured before and after administration of agents. Immunofluorescence double staining for CD31 and alpha-SMA was performed, and a medical video endoscopy system with narrowband illumination (NBI) was used to visualize the vascular pattern. RESULTS: TSU68 treatment decreased IFP significantly. Immunofluorescence double staining showed a significant increase in the fraction of pericyte coverage in the TSU68-treated group. NBI endoscopy showed that many tumor vessels in TSU68-treated mice were pruned and the diameters of remaining vessels were reduced. CONCLUSION: The data supported our hypothesis of tumor vascular normalization by the antiangiogenic agent TSU68. PMID- 19997801 TI - Changes in diagnostic strategies for hilar-type Castleman's disease: report of two cases. AB - Castleman's disease (CD) is rare and difficult to diagnose preoperatively. We report two similar cases of hyaline vascular type CD diagnosed preoperatively using different modalities. In the first case, a biopsy specimen taken during diagnostic thoracoscopy revealed lymphatic structure suggestive of CD. In the second case, endobronchial ultrasound-guided transbronchial needle aspiration enabled sampling of histological cores for histological diagnosis. In both cases, the final diagnosis was established by surgical resection. PMID- 19997800 TI - Effect of thrombopoietin on platelet counts and liver regeneration after partial hepatectomy in a rat model. AB - PURPOSE: We investigated the effects of thrombopoietin (TPO) on platelet counts and liver regeneration after partial hepatectomy in rats. METHODS: We performed 70% partial hepatectomy on 60 rats given either TPO or a vehicle (controls). The rats were killed 6 h, 24 h, 72 h, or 168 h after the procedure, and we examined the platelet counts, weight of the regenerated liver, hepatocyte proliferation by bromodeoxyuridine (BrdU) labeling index, and expression of hepatocyte growth factor (HGF) mRNA by reverse transcription-polymerase chain reaction. RESULTS: In the controls, the platelet counts were significantly lower than the basal levels, by about 20%, at 6, 24, and 72 h; then recovered to the basal level at 168 h. Conversely, in the TPO-treated rats, the platelet counts increased significantly by 17%, 35%, and 60%, at 24, 72, and 168 h, respectively. The regenerated liver regained 80% of the pre-hepatectomy weight by 72 h in the controls, whereas reconstitution of the liver was accomplished by 72 h in the TPO-treated rats. Thrombopoietin significantly enhanced the BrdU labeling index of hepatocytes and expression of HGF mRNA 24 h post hepatectomy in TPO-treated rats versus controls. CONCLUSION: Thrombopoietin increased platelet counts; thereby accelerating liver regeneration after partial hepatectomy with enhanced induction of HGF. PMID- 19997802 TI - Chondroblastic osteosarcoma arising from the pleura: report of a case. AB - Extraskeletal osteosarcoma is an uncommon malignant neoplasm. The origin of osteosarcoma in the pleura is extremely rare, with only four such cases so far documented in the literature to the best of our knowledge. We herein report the case of a 64-year-old Japanese man in whom a left pneumonectomy and pleurectomy were carried out to remove a huge tumor. The pathological examination confirmed a diagnosis of chondroblastic osteosarcoma that had originally arisen from the pleura. PMID- 19997803 TI - Successful treatment of acute esophageal necrosis caused by intrathoracic gastric volvulus: report of a case. AB - Gastric volvulus is a potentially lethal condition. We report a case of esophageal hiatal hernia with strangulation of the esophagus and stomach caused by gastric volvulus. A 79-year-old woman was admitted to our hospital in a state of shock, and investigations showed necrotic changes in most of her distal esophagus and gastric body. Thus, we performed an emergency total gastrectomy and transhiatal esophagectomy, followed 3 months later by successful reconstruction of the esophagus using the jejunum. Occasionally, a large hiatal hernia accompanies gastric volvulus; however, the extent of esophageal necrosis observed in this patient is very unusual. Although a large hiatal hernia is usually a chronic disorder, surgical treatment is recommended, considering the risk of serious complications. PMID- 19997804 TI - Gastric metastasis of malignant fibrous histiocytoma diagnosed preoperatively by endoscopic ultrasound-guided fine-needle aspiration biopsy: report of a case. AB - Metastatic gastric tumors originating from malignant fibrous histiocytoma (MFH) are rare. We herein report the case of a 75-year-old woman who underwent a distal gastrectomy for gastric metastasis of MFH, who was preoperatively diagnosed by an endoscopic ultrasound-guided fine-needle aspiration biopsy. PMID- 19997805 TI - Post-transplant lymphoproliferative disorder localized in the colon after liver transplantation: report of a case. AB - Post-transplant lymphoproliferative disorder (PTLD) is a life-threatening complication of any organ transplantation. It is more common in children than in adults, and the risk factors include Epstein-Barr virus (EBV) infection and immunosuppression. We report a case of colonic marginal zone B-cell lymphoma occurring 4 years after liver transplantation in a 52-year-old man who had been taking immunosuppressive agents, namely, cyclosporin, mycophenolate mofetil (MMF), and prednisolone. A routine follow-up colonoscopy showed a 2.0-cm diffuse nodular transverse colon polyp covered with erosive mucosa. Snare polypectomy was done without any complications. Immunohistochemical staining was positive for CD3 and CD20 in the interstitially scattered small lymphocytes and negative for Bcl 2, revealing marginal zone B-cell lymphoma. EBV quantitative polymerase chain reaction was less than the detection limit (<10 copies/5 microl whole blood). Extraintestinal whole-body screening was negative. The patient underwent right hemicolectomy after colonoscopic tattooing for positive resection margins on snare polypectomy. Surgical pathology revealed no evidence of residual lymphoma. Treatment consisted of colonic resection and reduction of the doses of immunosuppressants. The patient is currently under close surveillance without any signs of recurrence. PMID- 19997806 TI - Ileal J-pouch perforation at the blind end: report of a case. AB - A 34-year-old woman presented with severe abdominal pain 8 years after undergoing ileal J-pouch anal anastomosis for ulcerative colitis (UC). Computed tomography (CT) showed free air and ascites in the abdomen, so she underwent laparotomy. A perforation was found at the ileal J-pouch blind end, which was remarkably enlarged. When we resected the blind end surgically, we observed a 3-mm perforation and some small ulcers. About 2 months after this operation, the patient complained of abdominal pain and an increased frequency of bowel movements. Based on our endoscopic observation of multiple ulcers, we diagnosed pouchitis. We report this case to show that the combination of an enlarged J pouch blind end and pouchitis can result in perforation. To prevent such complications, we recommend avoiding the formation of a blind end during construction of a J-pouch. PMID- 19997807 TI - Early infection of peripancreatic tissue in mild acute pancreatitis: report of a case. AB - Mild acute pancreatitis (AP) is rarely complicated by infection, and the value of prophylactic antibiotics is questionable. We report a case of mild AP complicated by infection, which developed within 1 week after the onset. A 66-year-old woman was referred to our hospital where a diagnosis of mild AP was made, based on laboratory data and computed tomography (CT) findings. She was managed conservatively with fluid resuscitation, intravenous antibiotics, and protease inhibitor. Her general condition improved initially, but a high fever redeveloped on hospital day 3. On hospital day 7, a repeat CT scan showed a peripancreatic fluid collection with gas, indicating peripancreatic abscess. A drainage operation was performed, and the organism cultured from the abscess was Escherichia coli. Her postoperative course was uneventful. We report this case to stress that infection may develop even in mild AP, and even in the early phase. PMID- 19997808 TI - Pancreaticojejunostomy with invagination of the punched pancreatic remnant after medial pancreatectomy and enucleation for multiple metastases of renal cell carcinoma: report of a case. AB - We report the successful resection of multiple pancreatic metastases of renal cell carcinoma (RCC), achieved by performing medial pancreatectomy and enucleation, preserving as much of the pancreatic parenchyma as possible. Most of the distal remnant pancreas was placed into the jejunal lumen and all three cut surfaces were covered to prevent pancreatic leakage. The postoperative course was uneventful, without any sign of pancreatic fistula. The patient is well without any evidence of recurrence or impairment of exocrine or endocrine pancreatic functions 1 year after surgery. Considering the unusual behavior of RCC metastasis and the difficulty in predicting the pattern of recurrence, we should devise the optimal surgical strategy to provide cancer-free surgical margins and preserve as much of the pancreatic parenchyma as possible. PMID- 19997809 TI - Resection of four synchronous invasive ductal carcinomas in the pancreas head and body associated with pancreatic intraepithelial neoplasia: report of a case. AB - This report describes a very rare case of four synchronous invasive ductal carcinomas (IDCs) in the pancreas head and body with possible multicentricity. The patient was a 75-year-old woman. Abdominal dynamic computed tomography showed four low-density masses (25 mm, 20 mm, 10 mm, and 10 mm in diameter) in the pancreas head and body. The patient underwent a pylorus-preserving subtotal pancreatoduodenectomy. Histologically, the discontinuity between the four tumors was confirmed; one tumor (20 mm) was moderately differentiated tubular adenocarcinoma, and the others (25 mm, 10 mm, and 10 mm) were papillary adenocarcinomas. Two smaller papillary adenocarcinomas were composed of abundant fibrosis, pancreatic intraepithelial neoplasia (PanIN) 2-3, and IDC with stromal invasion. PanIN-1-2 lesions proximal to the four IDCs were evident. The immunohistochemical staining by CK20, MUC1, and Ki-67 revealed apparently different features for 2 IDCs (25 mm and 20 mm) and somewhat differential features for three papillary adenocarcinomas. Therefore, the progression of PanIN to IDC and multicentric occurrences of these four IDCs were possible. In this report, we show that immunohistochemistry and the confirmation of the presence of PanINs in IDC were useful to some extent for the study of multiple pancreatic cancers. PMID- 19997810 TI - Laparoscopic treatment of splenosis: report of a case. AB - A 52-year-old woman with a history of splenectomy for a splenic injury 45 years earlier underwent investigations for a pre-existing gynecologic disease. Contrast enhanced abdominal computed tomography (CT) and magnetic resonance imaging (MRI) showed a tumor, 3 cm in diameter, adjacent to the sigmoid colon, suggestive of a gastrointestinal stromal tumor (GIST). Laparoscopic exploration revealed a white mass protruding from the subserosa of the sigmoid colon, and we performed laparoscopic-assisted sigmoidectomy to rule out a GIST. Pathological studies showed splenic tissue, thus confirming a diagnosis of splenosis, given the history of splenic injury. This case of splenosis mimicking GIST following splenic injury was diagnosed correctly and treated successfully with laparoscopic surgery. To our knowledge, this is the first case of splenosis treated with laparoscopic colectomy with no tumor extirpation. PMID- 19997811 TI - Laparoscopic distal pancreatic resection: our own experience in the treatment of solid tumors. AB - A laparoscopic resection is a new treatment for pancreatic tumors. Articles by surgeons who are writing about their first experience in carrying out this treatment have appeared in the literature, reporting that laparoscopic surgery can be used for the treatment of pancreatitis, benign lesions, and solid tumors. This is a study of three patients with pancreatic tumors who were treated by means of a laparoscopic distal pancreatic resection with preservation of the spleen and splenic vessels. In three cases a laparoscopic distal resection was performed for the tumor. The histologic examinations showed one insulinoma and two mucinous cystadenomas. No patient suffered from intra- or postoperative complications. A laparoscopic resection of the distal pancreas is a new alternative for the treatment of pancreatic tumors. This method takes advantage of the benefits of minimally invasive surgery. PMID- 19997812 TI - Computed tomography is useful for preoperative workup of gastric rupture caused by blunt trauma. PMID- 19997813 TI - Association of low dietary vitamin K intake with radiographic knee osteoarthritis in the Japanese elderly population: dietary survey in a population-based cohort of the ROAD study. AB - BACKGROUND: The present study sought to identify dietary nutrients associated with the prevalence of radiographic knee osteoarthritis (OA) in the Japanese elderly of a population-based cohort of the Research on Osteoarthritis Against Disability (ROAD) study. METHODS: From the baseline survey of the ROAD study, 719 participants >or=60 years of age (270 men, 449 women) of a rural cohort were analyzed. Dietary nutrient intakes for the previous 1 month were assessed by a self-administered brief diet history questionnaire. The radiographic severity at both knees was determined by the Kellgren/Lawrence (KL) system. RESULTS: The prevalence of knee OA of KL >or=2 was 70.8%. Age, body mass index, and female sex were positively associated with the prevalence. Among the dietary factors, only vitamin K intake was shown to be inversely associated with the prevalence of radiographic knee OA by multivariate logistic regression analysis. The presence of joint space narrowing of the knee was also inversely associated with vitamin K intake. The prevalence of radiographic knee OA for each dietary vitamin K intake quartile decreased with the increased intake. CONCLUSIONS: The present cross sectional study using a population-based cohort supports the hypothesis that low dietary vitamin K intake is a risk factor for knee OA. Vitamin K may have a protective role against knee OA and might lead to a disease-modifying treatment. PMID- 19997814 TI - Bone mass assessment in naval crew members by quantitative ultrasound technique. AB - OBJECTIVE: Of necessity, naval crews live in confined spaces when on board warships, which may lead to decreased bone mass and to subsequent bone fractures. Therefore, we investigated the bone mass of crew members and the relationship between bone mass and lifestyle factors. METHODS: We selected 1510 crew members of the Japan Maritime Self-Defense Force. All were men between 18 and 58 years of age. We measured their bone mass by applying quantitative ultrasound (QUS) to the calcaneus. In addition, we reviewed daily milk consumption, levels of physical exercise, type of on-board job, fracture history, nutritional supplementation habits, and body mass index (BMI). RESULTS: Bone mass values were lower than the published mean values for Japanese men across the twenties to forties age groups. CONCLUSIONS: The factors found to be related to bone mass in this study were age, type of on-board job, exercise level, and milk consumption. Lifestyle factors are usually within our control. Our data suggest that moderate levels of regular exercise and milk consumption may maintain bone mass. PMID- 19997815 TI - Clinical study of chronic lateral ankle instability: injured ligaments compared with stress X-ray examination. AB - BACKGROUND: A comparative analysis of ankle stress X-ray findings and the condition of injured ligaments in patients with chronic lateral ankle instability was performed to determine a method of identifying the condition of injured ligaments prior to operation. METHODS: Altogether, 36 males and 80 females were included in the study. The average age at the time of operation was 29.4 years. Anterior drawer distances and talar tilt angles were measured manually. During the operations, the injured ligaments were classified into 25 categories. RESULTS AND CONCLUSION: The condition of the injured ligaments in patients with a talar tilt angle of more than 15 degrees had injured ligaments that were almost avulsed and degenerated or completely absent. PMID- 19997816 TI - Antibiotics-impregnated cement spacers in the first step of two-stage revision for infected totally replaced hip joints: report of ten trial cases. AB - BACKGROUND: Periprosthetic infection is one of the serious complications after total hip arthroplasty (THA). This study analyzed the perioperative and postoperative status of patients who underwent antibiotics-impregnated cement spacer technique in the first step of the two-stage revision. METHODS: Ten joints of the nine patients (mean age, 65 years; seven women, two men) received two stage revision as a result of infection that appeared after primary THAs in seven joints, aseptic revision in one, and recurrent type in two. An antibiotics impregnated cement spacer made by a mold system was applied in the femoral side of all joints. An acetabular spacer was made by hand using a cup gauge in eight joints with extensive tissue loss. RESULTS: The change of leg length after the first stage was -2.2 mm, and range of hip flexion was 72 degrees on average, respectively. Patients could walk with crutches after the first stage, except one patient with simultaneous infections of both hips and one with fracture of the cement spacer. One fracture of femoral cement spacer, and one dislocation of femoral spacer accompanied by fracture of acetabular cement spacer and curable recurrent infection, were found. In all cases of the second-stage procedure, the acetabular side was reconstructed with allogeneic bone graft with cross plate and that of the femur was by impaction bone grafting method. In the latest follow-up, reconstructed implants were stable. Seven patients could walk without any supportive devices and two could walk with the support of a T-cane. CONCLUSIONS: An antibiotics-impregnated cement spacer in the first step of the two-stage revision was effective not only to compensate tissue loss after removal of the implants and to minimize discrepancy of leg length, but also to contribute to improvement of perioperative and postoperative daily activities of the patient's life as well as treatment of the infection. PMID- 19997817 TI - Validation study of a clinical diagnosis support tool for lumbar spinal stenosis. AB - BACKGROUND: A clinical diagnosis support tool for lumbar spinal stenosis was developed by the Japanese Society for Spine Surgery and Related Research. However, the use of this tool has not yet been validated. METHODS: Patients with symptoms in the lower extremities and who visited the Department of Orthopedics initially were recruited to the study. Orthopedic physicians who were not spine specialists completed the support tools. Spine specialists examined the patients, made a diagnosis, and completed the lumbar spine examination sheet made for the study. The support tool and lumbar spine examination sheet were sent to a central panel comprising four panelists who then decided on a final diagnosis. RESULTS: In total, 118 patients were evaluated, including 62 males and 56 females. Lumbar spinal stenosis was diagnosed in 58 and nonlumbar spinal stenosis in 60 patients. The mean score in the lumbar spinal stenosis group was 12.2 points (median 13 points). In the nonlumbar spinal stenosis group, the mean score was 7.5 points (median 7 points). Sensitivity was 0.948, and specificity was 0.40. CONCLUSIONS: Patients with lumbar spinal stenosis with a very low score were diagnosed with mild lumbar spinal stenosis, whereas nonlumbar spinal stenosis patients with a very high score were diagnosed as suffering from spine disease and needing special treatment by spine surgeons. Our results validate the use of the support tool for the diagnosis of lumbar spinal stenosis. Although the specificity observed in the present study was lower than that reported at development, we conclude that this support tool is useful for screening patients with lumbar spinal stenosis. PMID- 19997818 TI - Cement removal and bone bed preparation of the femoral medullary canal assisted by flexible endoscope in total hip revision arthroplasty. AB - PURPOSE: The flexible endoscope was applied to cemented femoral medullary canal to obtain better visual field and assist surgical procedures in total hip revision arthroplasty. METHODS: Fifteen cases of failed cemented total hip joints were analyzed. Efficacy of cement removal was assessed, combined with degree of implant loosening and bone defect, postoperative radiographic findings, and perioperative status of the patients and complications. Status of the bone bed between bone and cement and that of bone grafting were also evaluated. RESULTS: The cement mantle was efficiently extractable in all cases under good exposure and with maintenance of efficient working space. Endoscopic time for cement removal was dependent on the status of the cement-bone interface and bone defect. Rigid and less loosened interfaces, as well as cases of minimal bone defect, required a longer time. However, it was effective to confirm the status of the bone bed during the procedure. Occult foreign-body reaction was detectable in three cases of unloosened interface under endoscopic inspection. Impaction bone grafting was performed in eight cases. The scope was also helpful to confirm the status of a grafted bone bed. Three fractures occurred, of which two cases revealed minor cement leakage and one required additional osteosynthesis with extensive approach. JOA Hip Score was improved, and the implants were stable at latest follow-up. Dislocation was found in two cases. Neither thromboembolic events nor infection was found. CONCLUSIONS: Cement removal in the femoral medullar canal was effectively performed not only in the cases of loose interface but also those of rigid and less-loosened interface under flexible endoscopic inspection. The scope could support cement removal in femoral revisions under good exposure and maintenance of working space, as well as confirmation of bone bed state. Although one case required an additional approach, application of the flexible endoscope has potential merit to contribute to less invasive total hip revision arthroplasty, possibly combined with other refined devices of cement extraction. PMID- 19997819 TI - Total hip arthroplasty after hip joint ankylosis. AB - BACKGROUND: Back pain and knee pain are typical secondary degeneration symptoms after hip ankylosis. Take down of hip ankylosis and implantation of a total hip arthroplasty (THA) is believed to be a promising treatment option. METHODS: A total of 22 hip ankylosis patients [15 men, mean age 53.7 years (range 30-72 years); 7 women, mean age 50.8 years (range 42-61 years)] underwent THA during 1980-2000 after spontaneous (n = 10) or surgical (n = 12) fusion of the hip joint. The mean duration of ankylosis prior to THA was 32.5 years (range 2-61 years). RESULTS: At the mean follow-up of 13.2 years (range 2-19 years), the Harris hip score averaged 84.9 points (range 70.1-99.0 points). All patients (100%) confirmed that they would undergo conversion surgery again. Aseptic loosening of two stems (one cemented, one cementless; 9.5%) and two deep infections (9.5%) required revision surgery. CONCLUSIONS: THA is a promising option for treatment of secondary long-term hip ankylosis sequelae. A conversion operation after spontaneous ankylosis provides better functional outcome than after surgical fusion. However, full function with complete pain relief and a negative Trendelenburg sign might be not attainable in all cases. PMID- 19997820 TI - Immunolocalization of lactoferrin in cartilage-forming neoplasms. AB - BACKGROUND: Lactoferrin (Lf) is an 80-kDa basic glycoprotein, a member of the transferrin family of iron-binding proteins. Lf immunoreactivity has been extensively investigated in many neoplastic tissues. Recently, Lf expression was documented in the osteoblastic lineage of bone-forming tumors as well as in osteoblasts of fetal bone. METHODS: Lactoferrin (Lf) immunoexpression was investigated in 30 human cartilage-forming tumors [15 enchondromas, 6 osteochondromas, 3 chondroblastomas (CBL), 3 chondrosarcomas, and 3 chondromyxoid fibromas (CMF)] as well as in human normal bone specimens and cartilaginous tissues obtained at autopsy from 5 adults and 3 fetuses.In addition, the immunohistochemical expression of Ki-67 antigen was analyzed on parallel sections from the same specimens. Quantification of Lf immunoreactivity was performed by using an intensity distribution (ID) score. RESULTS: Lf immunoexpression with a variable ID score was encountered exclusively in 3 of 3 chondroblastomas and in 3 of 3 chondromyxoid fibromas. Lf immunoreactivity in these tumors, in clear contrast with the Lf absence in enchondromas, osteochondromas, and chondrosarcomas, may suggest a different histogenesis of the former. In agreement with this histogenetic origin, we detected Lf in the chondroblasts and osteoblasts within the fetal tissue, whereas no immunoreactivity was found in the corresponding adult cells. No significant associations were found between the Lf immunoexpression and the Ki 67 LI of the tumors of our series. CONCLUSIONS: The presence of Lf in neoplastic cells of CBL and CMF, as well as in fetal cartilaginous tissue, may reflect a less mature phenotype of these tumors. PMID- 19997821 TI - Asporin and transforming growth factor-beta gene expression in osteoblasts from subchondral bone and osteophytes in osteoarthritis. AB - BACKGROUND: To clarify the significance of subchondral bone and osteophytes in the pathology of osteoarthritis (OA), we investigated the expression of asporin (ASPN), transforming growth factor-beta1 (TGF-beta1), TGF-beta2, TGF-beta3, and runt-related transcription factor-2 (Runx2) genes involved in bone metabolism. METHODS: Osteoblasts were isolated from 19 patients diagnosed with knee OA and from 4 patients diagnosed with femoral neck fracture. Osteoblast expression of mRNA encoding ASPN, TGF-beta1, TGF-beta2, TGF-beta3, and Runx2 was analyzed using real-time RT-PCR. RESULTS: Expression of ASPN, TGF-beta1, and TGF-beta3 mRNA in the subchondral bone and osteophytes of OA patients increased compared with that of non-OA patients. The ratio of ASPN to TGF-beta1 mRNA in patients with severe cartilage damage was higher than that in patients with mild cartilage damage. CONCLUSIONS: The increased ratio of ASPN mRNA to TGF-beta1 mRNA in patients with severe relative to mild cartilage damage indicates that increased ASPN mRNA expression was significantly associated with the severity of cartilage degeneration. This finding suggests that ASPN may regulate TGF-beta1-mediated factors in the development of OA, which may provide clues as to the underlying pathology of OA. PMID- 19997822 TI - Changes in the sagittal spinal alignment of the elderly without vertebral fractures: a minimum 10-year longitudinal study. AB - BACKGROUND: Thoracic hyperkyphosis, or loss of lumbar lordosis, is often equated with osteoporosis because vertebral fractures are assumed to be a major causative factor. However, recent evidence suggests that up to one-half of the patients with hyperkyphosis have no evidence of underlying vertebral fracture. The shape characteristics of the intervertebral discs and their role in determining kyphotic curvature have been investigated. The spinal sagittal parameters and segmental disc angles of elderly subjects were examined during a longitudinal follow-up. METHODS: A total of 53 subjects (20 men, 33 women) without vertebral fractures during a more than 10-year follow-up were included in this study, undergoing standing lateral radiographs of the spine using 36-inch film at baseline and final follow-up. The mean age of the subjects was 63 years (range 50 77 years) at baseline and 75 years (range 62-88 years) at follow-up; and the mean follow-up period was 11 years 11 months. RESULTS: The lumbar lordosis and the sacral inclination angle decreased and the C7-plumbline distance increased with age. Among a total of 664 discs, 70 discs (10.5%) showed anterior wedging over 5 degrees at follow-up. In contrast, 39 discs (5.9%) showed posterior wedging over 5 degrees . The subjects had only discs with anterior wedging, decreased total lumbar lordosis, and the C7 plumbline displaced anteriorly. However, when the subjects had discs with posterior wedging, the C7 plumbline and sagittal spinal balance tended to be maintained. This compensatory mechanism was seen in younger subjects. CONCLUSIONS: A decrease in the total lumbar lordosis and the sacral inclination angle occurred with age. Increasing age correlated with a more forward sagittal vertical axis, depending on a decrease in the total lumbar lordosis. The cause of loss of lumbar lordosis in the subjects without vertebral fracture was anterior wedging of the segmental discs. Posterior wedging of the thoracic and lumbar segmental discs then could occur, compensating for the loss of lumbar lordosis. PMID- 19997823 TI - The effect of geometry of the tibial polyethylene insert on the tibiofemoral contact kinematics in Advance Medial Pivot total knee arthroplasty. AB - BACKGROUND: In modern total knee arthroplasty (TKA), it is important to reproduce both medial pivot motion and posterior femoral rollback to obtain greater postoperative knee flexion. Several studies have reported the factors affecting knee motion and range of motion after TKA. The purpose of this study was to evaluate the effect of the tibial insert geometry on the tibiofemoral contact kinematics, especially focusing on the medial pivot motion and posterior femoral rollback. METHODS: Seven cadaveric knees were replaced with the Advance Medial Pivot TKA, and two different geometries of polyethylene tibial insert, the standard medial pivot design (MP-design) and double high design (DH-design), were biomechanically compared. Four experimental configurations were evaluated in each specimen in this order: (1) the MP-design with posterior cruciate ligament (PCL) retaining, (2) the DH-design with PCL retaining, (3) the MP-design with PCL sacrificing, and (4) the DH-design with PCL sacrificing. RESULTS: Under the PCL retaining condition, both designs showed no medial pivot but bicondylar femoral rollback more than 60 degrees of knee flexion. In the MP-design, tibiofemoral contact point (estimated contact point, ECP) of the medial compartment was located on the posterior lip of the ball-insocket structure while demonstrating greater than 120 degrees of knee flexion. The posterior translation was also the same in both designs. On the other hand, ECP of the MP-design and the DH-design showed only medial pivot pattern under the PCL-sacrificing condition. In the DH design, ECP of the lateral compartment showed paradoxical anterior translation from 0 degrees to 60 degrees of knee flexion. Total posterior translation was significantly greater in the lateral compartment than that in the medial compartment. CONCLUSIONS: The results of this study suggest that in this type of TKA system the ball-in-socket geometry in the MP-design has an advantage for reproducing medial pivot motion in the PCL-sacrificing condition, and the flexion path structure in the DH-design is considered to be both effective and safe for femoral rollback in the PCL-retaining condition. However, neither design is sufficient to reproduce medial pivot motion and posterior femoral rollback. Therefore, a different design of tibial insert is needed for more physiological kinematics after TKA. PMID- 19997824 TI - Histological examination of frozen autograft treated by liquid nitrogen removed after implantation. AB - BACKGROUND: Several oncological sterilization methods involving autoclaving, irradiation, or pasteurization have been developed for limb reconstruction of large bone defects following tumor excision. Studies involving histological examinations of these autografts have all found that osteogenesis occurs slowly. We have used frozen autografts treated by liquid nitrogen for limb reconstruction and have achieved excellent results for bone union. To determine if frozen autografts exhibit early bone remodeling, we investigated the repair processes of the frozen bones. METHODS: We analyzed frozen autografts treated by liquid nitrogen, retrieved at a mean of 19.1 months (2-75 months) after implantation because of complications or local tumor recurrence. The specimens were obtained from six patients with a mean age of 36.2 years (8-68 years). The six grafts comprised three osteoarticular grafts, two intercalary grafts, and one joint graft. We histologically reviewed the autograft-containing sections for tumor cell necrosis, evidence of cortical repair, the cortical junction, and joint cartilage. RESULTS: Tumor cells were completely eradicated from the frozen bone in all cases. In a specimen retrieved 5 months after implantation, a small area of the bone showed active osteocytes and osteoblasts. In three cases retrieved more than 1 year after implantation, osteocytes and osteoblasts were observed in broad portions of the frozen bones, indicating the onset of osteogenesis in the frozen bone at an early stage. The cortical host-graft junction showed incorporation along with continuity of bone trabeculae. In addition, we were able to fi nd normal chondrocytes on the articular surface. CONCLUSIONS: The frozen bone specimens in this study thus showed evidence of newly formed bone and earlier osteogenesis than has been previously reported. Our results suggest that frozen autografts may be considered one of the most useful recycled materials for biological reconstruction. PMID- 19997825 TI - Quantitative analysis of Staphylococcus epidermidis biofilm on the surface of biomaterial. AB - BACKGROUND: Staphylococcus epidermidis biofilm is considered to be an important cause of device-related infection. Polysaccharide intercellular adhesin (PIA), encoded by the icaADBC locus, has been found to be a functional component of S. epidermidis biofilm, but the sequential change of the ica gene expression during biofilm development is still unclear. We have established a quantitative experiment of biofilm formation on nontranslucent biomaterial surfaces using the biofilm coverage rate (BCR). In this study, we quantified the time course of biofilm formation on a biomaterial (stainless steel) surface by means of BCR, viable cell count (VCC) with colony-forming units, and ATP-bioluminescence (ATP) as relative light units, and investigated the time-course relationship between biofilm development process and ica gene expression using reverse transcription polymerase chain reaction (RT-PCR). METHODS: S. epidermidis RP62A was inoculated on stainless steel washers and incubated for 0-8, 24, and 48 h. Biofilms attached to the washers were quantified by means of BCR, VCC, and ATP. RT-PCR of the ica gene was performed using total RNA isolated from biofilms at each incubation period. Results of these methods were compared. RESULTS: The amount of biofilms measured by BCR increased over time and particularly grew at 5-6 h into the incubation period. On the other hand, the results of VCC and ATP increased gradually, and at 24 h or 48 h the measurement values were very much greater than previously. Up to 8 h, there were significant correlations between BCR and VCC or ATP. The growth of BCR until 6 h is supported by RT-PCR of the ica gene. CONCLUSIONS: Compared with each result, two-dimensional biofilm occupation on a biomaterial surface is proposed to be rapidly completed within 6-8 h after bacterial attachment. Our data indicate that bacterial biofilms first grow two dimensionally with a producing matrix, and subsequently grow vertically and become mature. PMID- 19997826 TI - Influence of extracellular matrix on the expression of inflammatory cytokines, proteases, and apoptosis-related genes induced by hydrostatic pressure in three dimensionally cultured chondrocytes. AB - BACKGROUND: The purpose of this study was to investigate the influence of hydrostatic pressure (HP) on the gene expression of cartilage matrix, cytokines, and apoptosis-associated factors in chondrocytes in which the cartilage was in extracellular matrix (ECM)-rich or ECM-poor condition. METHODS: Chondrocytes were isolated from rabbit joints and cultured in alginate beads. Immediately after embedding (0W group) or after 2 weeks culture (2W group), the amounts of glycosaminoglycan (GAG) in the alginate beads were quantified. Both groups were exposed to continuous HP of 10 or 50 MPa for 12 h. The expression of inflammatory cytokines, proteases, and apoptosis-related factors were examined by reverse transcription-polymerase chain reaction (RT-PCR). The expression of proteoglycan core protein (PG) and collagen type II were quantified by real-time RT-PCR. RESULTS: All of the GAG components in alginate beads markedly increased in the 2W group. The expression of PG and collagen type II increased after exposure to 10 MPa in both groups. In the 0W group, these levels decreased after exposure to 50 MPa of HP. The expression of interleukins IL-6 and IL-8 increased after exposure to HP in the 0W group. HP at 50 MPa induced mRNA expression of ADAMTS-5 in the 0W group but not in the 2W group. The expression of Fas increased after exposure to HP in the 0W group. CONCLUSIONS: These findings suggested that nonphysiological, excessive HP on chondrocytes with the ECM in poor condition reduced matrix gene expression and increased expression of the genes associated with apoptosis and catabolism of the cartilage matrix. These results might therefore be associated with the pathogenesis of osteoarthritis. PMID- 19997827 TI - In vitro comparison of elution characteristics of vancomycin from calcium phosphate cement and polymethylmethacrylate. AB - BACKGROUND: Calcium phosphate cement [CPC (Biopex)] has been used as the drug delivery system of choice for treatment of infected joint replacement because of its good elution efficiency. The influence of CPC polymerization on the bactericidal activity of vancomycin (VCM) impregnated into CPC has not been investigated. We compared VCM concentration, bactericidal activity, and profile of eluates between CPC and polymethylmethacrylate (PMMA; Cemex RX). METHODS: Test specimens consisted of a powder composite of CPC or PMMA, VCM and solvent (10:0.25:3.3 g). Each test specimen was immersed in sterile phosphate-buffered saline. Eluates obtained on days 1, 3, 7, and 14 and weeks 4, 8, and 12 were evaluated by high performance liquid chromatography (HPLC) and by microbiological assay (MBA). RESULTS: The elution level of VCM from CPC/VCM on day 1 was 8.1 fold greater than that from PMMA/VCM. The detection periods of VCM from CPC/VCM and PMMA/VCM were 8 weeks and 14 days, respectively. The values of eluates from CPC/VCM and PMMA/VCM obtained by HPLC were comparable to those obtained by MBA. HPLC chromatogram showed that the elution profiles of VCM from CPC/VCM and PMMA/VCM on day 1 were very close to those of standard solutions. CONCLUSIONS: CPC could release more VCM over a longer period than PMMA. The polymerization of CPC and PMMA did not alter the inhibitory activity of VCM and did not denature VCM. PMID- 19997828 TI - Low levels of steroid-metabolizing hepatic enzyme (cytochrome P450 3A) activity may elevate responsiveness to steroids and may increase risk of steroid-induced osteonecrosis even with low glucocorticoid dose. AB - BACKGROUND: The main purpose of this study was to examine the relationships among osteonecrosis, steroid-metabolizing hepatic enzyme (cytochrome P450 3A; CYP3A) activity, and steroid dose to determine whether it is possible to prevent osteonecrosis in animals with low hepatic CYP3A activity by reducing exogenous steroid doses. METHODS: Japanese white rabbits (n = 103) were divided into three groups: a group with CYP3A activity induction (by intramuscular phenobarbital injection, n = 31), a group with CYP3A activity inhibition (by oral itraconazole administration, n = 30), and a control group (n = 42). Three weeks later, all rabbits received a methylprednisolone injection. Each group was divided into two subgroups by dosage of methylprednisolone (5 or 10 mg/kg body weight). Three weeks after methylprednisolone injections, the animals were killed and histological examination was performed to determine the incidences of osteonecrosis in the six subgroups. RESULTS: Incidence in the inhibition subgroup with 5 mg/kg steroid was higher than that in the induction subgroup receiving 10 mg/kg steroid. Thus, suppression of CYP3A activity significantly increased vulnerability to steroid-induced osteonecrosis, while increased CYP3A activity reduced this vulnerability. CONCLUSIONS: These findings suggest that low CYP3A activity may be vulnerable to the effect of steroids and increase risk of osteonecrosis, even with a low dose of steroid. PMID- 19997829 TI - Local application of olprinone for promotion of peripheral nerve regeneration. AB - BACKGROUND: Olprinone was applied locally to the sciatic nerves of rats, and nerve regeneration activity was examined in relation to heat shock protein (HSP)27 expression. METHODS: Adult rat sciatic nerves were locally exposed to olprinone, physiological saline, or a mixture of saline and HCl and underwent crush injury. HSP27 protein levels were assessed by Western blotting and immunohistochemistry. Nerve regeneration was assessed by sciatic function index (SFI) and by morphological evaluation. RESULTS: Olprinone was detected in ipsilateral L4-L6 dorsal root ganglion (DRG) and sciatic nerve proximal to the site of crush injury. Western blot analysis of the DRG 24 h postsurgery and the sciatic nerve 7 days postsurgery showed a marked increase in HSP27 protein levels in the olprinone-treated group. Immunohistochemical analysis was consistent with the Western blot analysis. SFI was significantly better for the olprinone-treated group than for either the saline- or HCl-treated groups at 7, 22, and 25 days, respectively, after application. The packing density and the number of myelinated axons 21 days after the crush injury in the olprinone-treated group were significantly greater in comparison to the other groups. CONCLUSION: The local application of olprinone to the sciatic nerve increases HSP27 protein levels in DRG in the early stage and in the sciatic nerve 1 week thereafter. The results from this crush model indicate that increasing levels of HSP27 appear to result in therapeutic effects. PMID- 19997830 TI - Effects of single and cyclical local injections of basic fibroblast growth factor on cancellous bone defects in rabbits. AB - BACKGROUND: Local administration of basic fibroblast growth factor (bFGF) has anabolic effects on bone formation. A delivery system for local treatment is required to increase efficacy because of its short half-life. However, little is known about the effects of cyclical local injection of bFGF. We evaluated the effects of single and cyclical local injection of bFGF at a cancellous bone defect in the femoral condyle in rabbits. METHODS: Using the "vehicle only" as a control, a single low dose (40 microg), single high dose (120 microg), or cyclical low dose (40 microg, three times) of bFGF was injected percutaneously into a bone defect implanted with a gelatin sponge. The rabbits were killed at 4 weeks after surgery and the femurs were harvested for evaluation. RESULTS: Both single and cyclical administration of bFGF dose-dependently increased the amount of new bone formation in the bone defect using radiographs (P < 0.01) and bone mineral density (BMD) measurements (P < 0.01) compared to controls. However, only high-dose bFGF injection significantly increased the cancellous bone volume at the bone defect (P < 0.05) compared to controls, using bone histomorphometry. Cyclical injection of bFGF significantly increased the number of runt-related transcription factor-2 (Runx2)-positive cells compared to single low- and high dose bFGF administration (P < 0.01 and P < 0.05, respectively), and single high dose and cyclical administration significantly increased the number of osteopontin-positive cells compared to controls (P < 0.01), based on immunohistochemical analysis. CONCLUSIONS: These results suggest that high-dose injection of bFGF, at the very early stage of cancellous bone healing, is more effective in increasing cancellous bone volume, and cyclical injection of bFGF may stimulate osteoprogenitor cells. PMID- 19997831 TI - Monteggia fracture managed by intramedullary Kirschner wire fixation with pyknodysostosis. PMID- 19997832 TI - Extraarticular triplane fracture of the distal tibial epiphysis in a child. PMID- 19997833 TI - Glomus tumor of the toe with symptoms similar to those of Morton's disease. PMID- 19997834 TI - Total en bloc spondylectomy and a greater omentum pedicle flap for a large bone and soft tissue defect: solitary lumbar metastasis from renal cell carcinoma. PMID- 19997835 TI - Editorial. Some contemporary aspects within the field of Urology. PMID- 19997836 TI - [How to use PSA in 2009]. AB - PSA is without any doubt the most frequently used marker in urology due to its helpful information regarding various aspects of diagnosis, therapy and prognosis in men with prostate cancer. On the other hand, many controversies still exist about the various indications for PSA-determination. The following overview is aimed to analyse the current status of PSA in the management of men undergoing screening, therapy or follow-up of prostate cancer. Anyhow, a detailed knowledge of how to use and interpret PSA and PSA-kinetics is considered to play a crucial role in prostate cancer patients. Current strategies are aimed to start and stop PSA-use earlier. PMID- 19997837 TI - Prostate cancer in senior adults: over- or undertreated? AB - Despite the widespread use of prostate specific antigen for early prostate cancer (PCa) detection in younger men, PCa is still as disease of the elderly as 2/3 of incident cases are detected in men older than 65 years and 25% are older than 75 years at diagnosis. Opportunistic screening for PCa is not recommended for men with a life expectancy of less than 10 years. The therapeutic strategy for senior adults is driven by tumour stage/aggressiveness, co-morbidity and chronological age. Elderly patients with low/intermediate risk tumours - particularly those with a life expectancy of less than 10 years - are best managed by watchful waiting. Senior adults with intermediate/high risk tumours and a life expectancy of >10 years may benefit from curative local therapy such as radical prostatectomy or combined external beam irradiation/androgen ablation therapy. For elderly patients with metastatic disease, androgen deprivation remains the mainstay of therapy, intermittent androgen ablation is a promising approach. PMID- 19997838 TI - [Erectile dysfunction]. AB - Erectile Dysfunction is one of the most prevalent sexual disorders in men. According to the current literature the prevalence is about 16% for all men. The diagnostic workup of a patient suffering from E.D. is a detailed anamnesis, a physical and endocrinological evaluation and the suggestion of the optimal therapeutic treatment. The therapeutic application of PDE-5 inhibitors has made the therapy of E.D. much easier. It is a safe and highly efficient therapeutic option. But man must not forget that in severe cases of erectile dysfunction, the application of SKAT therapy, penile implants or vacuum devices might be necessary. Since a functioning sex life is important not only for the men, but also for the couple, a more open attitude of us, physicians, is desirable to help more men who suffer from this sexual dysfunction. PMID- 19997839 TI - [Current surgical aspects of renal cell carcinoma]. AB - Surgery of renal cell carcinoma (RCC) has undergone substantial changes during the past years. Until the 1990s, radical nephrectomy together with ipsilateral adrenalectomy was considered the golden standard. Meanwhile it was demonstrated that tumours up to 4 cm in diameter and a normal contralateral kidney can be treated by partial nephrectomy (elective indication) without compromising oncological safety. Development of laparoscopy influenced renal surgery significantly. Currently, laparoscopic nephrectomy is considered a golden standard for renal masses larger than 4 cm (T1b-T2), whereas open radical nephrectomy is reserved for large, locally advanced tumours, caval extension or enlarged lymph nodes. Adrenalectomy can be omitted if the adrenal appears normal. Tumours less than 4 cm should undergo partial nephrectomy. The better renal functional reserve after elective partial compared with radical nephrectomy seems to be associated with better overall survival. For partial nephrectomy the open approach currently remains the golden standard, whereas laparoscopy is reserved for specialized centres. PMID- 19997840 TI - [Chronic pain in urology: inguinal pain, post vasectomy pain, and chronic pelvic pain]. AB - Undertreatment of all forms of acute and chronic pain is also present in Germany. Chronic inguinal pain and post vasectomy pain are frequently observed in urological practice and appear as chronic postsurgical pain. Chronic pelvic pain has inflammatory causes in the majority of cases and appears as a challenge for the treating physician. Pain should be treated as early as possible, if necessary also interdisciplinary, with regard to the further chronification process with changes of biological, psychological and social factors. For the patient the pain chronification can be accompanied by the reduction of quality of life and social isolation. PMID- 19997841 TI - [Male-to-female transsexualism. Sex reassignment surgery from a biopsychosocial perspective]. AB - Many man-to-female-transsexuals prefer sex reassignment surgery. Surgical complications are common, most frequently a stenosis of the vagina as well as the urethral ostium. In up to 24% of patients secondary corrective surgery is necessary. Regret and feelings of doubt can occur in up to 8% of the cases. In the setting of an interdisciplinary team the postoperative somatic, psychological and socio-economic situation can be improved. This review is based on a Medline literature search and summarizes the pertinent literature of the last 22 years. PMID- 19997842 TI - [Achievement of blood lipid target levels with Ezetimibe/Simvastatin in patients with atherosclerosis and/or diabetes mellitus--an Austrian observational study]. AB - BACKGROUND: In patients at risk for cardiovascular complications, established LDL cholesterol (LDL-C) target levels are frequently not achieved using standard statin therapy regimens. The additional lipid-lowering efficacy of the fixed combination ezetimibe/simvastatin was evaluated in a countrywide non interventional observational study in Austria. METHODS: About 3,156 subjects with clinically manifest atherosclerosis and/or diabetes at high cardiovascular risk and with LDL-C-levels >113 mg/dl under statin therapy met the inclusion criteria and 2,903 of these patients were treated by primary care physicians, in hospitals and in rehabilitation centers with 10 mg ezetimibe and 10-40 mg simvastatin daily in a fixed-combination tablet (Inegy). RESULTS: During the follow-up period of 4 12 weeks, LDL-C levels were reduced by a median of 27-31% of baseline values (mean 153.1 +/- 33.5 mg/dl) mainly regardless of previous statin therapy (rosuvastatin, atorvastatin, simvastatin, pravastatin, fluvastatin, and lovastatin) and dosing (pooled median values). LDL-C reduction correlated proportional with baseline LDL-C values and increased with increasing simvastatin dosage. Overall, 45.3% (10/10 mg), 43.9% (10/20 mg) and 62.7% (10/40 mg ezetimibe/simvastatin) achieved LDL-C target levels of <100 mg/dl. CONCLUSIONS: The fixed combination therapy with ezetimibe/simvastatin showed a clinically significant additional lipid-lowering potential as compared with established statin monotherapies and enabled more patients at cardiovascular risk to reach the LDL-C target level of <100 mg/dl. PMID- 19997843 TI - Blockade of serotonin 2A receptors prevents PCP-induced attentional performance deficit and CREB phosphorylation in the dorsal striatum of DBA/2 mice. AB - RATIONAL AND OBJECTIVE: Functional opposition between N-methyl-D-aspartate and 5 HT(2A) receptors may be a neural mechanism supporting cognitive functions. These systems converge on an intracellular signaling pathway that involves protein kinase A-dependent phosphorylation of different proteins including cyclic adenosine monophosphate response element binding (CREB). Thus, we tested whether selective 5-HT(2A) receptor antagonist, M100907, might abolish phencyclidine (PCP)-induced attentional performance deficit by preventing its effects on transduction mechanisms leading to CREB phosphorylation. METHODS: Using the five choice serial reaction time task, the ability of subcutaneous injections of 2.5 and 10 microg/kg of M100907 to abolish the effects of an intraperitoneal injection of 1.5 mg/kg PCP on attentional performance as measured by accuracy (percentage of correct responses) and anticipatory and perseverative responding was assessed in DBA/2 mice. The effects of PCP, M100907, and their combination on S(133)-CREB and T(34)-DARPP32 phosphorylation in the dorsal striatum and prefrontal cortex (PFC) of behaviorally naive mice were examined using Western blotting technique. RESULTS: PCP reduced accuracy and increased anticipatory and perseverative responses as well as it increased S(133)-CREB phosphorylation in the dorsal striatum but not in the PFC. Ten microg/kg M100907 abolished the PCP induced attentional performance deficits and the increase in S(133)-CREB but not T(34)-DARPP32 phosphorylation. By itself, M100907 had no effect on attentional performance or phospho-S(133)-CREB and phospho-T(34)-DARPP32. Interestingly, the effect of PCP on phospho-S(133)-CREB but not on phospho-T(34)-DARPP32 was dependent on endogenous 5-HT. CONCLUSIONS: The data indicate that blockade of 5 HT(2A) receptors may exert beneficial effects on cognitive deficits through a mechanism linked to striatal S(133)-CREB phosphorylation. PMID- 19997845 TI - The regulatory action of alpha-actinin on actin filaments is enhanced by cofilin. AB - We have used fluorescence recovery after photobleaching to study the effect of muscle alpha-actinin on the structure of actin filaments in dilute solutions. Unexpectedly we found that alpha-actinin partitioned filaments into two types: those with a high mobility and those with low mobility. We have determined that the high mobility (smaller sized) population is too large to be simple monomeric actin:alpha-actinin complexes. Although it is known that cofilin encourages the transformation of alpha-actinin:actin gels into large meshworks of inter digitating actin filament bundles (Maciver et al. 1991), we have found that the presence of cofilin also increases the cross-linking of actin filaments by alpha actinin and hypothesize that this is due to cofilin's ability to alter the filament twist. This effectively makes more potential alpha-actinin binding sites per unit of actin filament. As expected from previous work, this effect was more marked at pH 6.5 than at pH 8.0. Both effects are likely to operate in cells to deny other actin-binding proteins access to binding these particular filaments and may explain how very different actin cytoskeletal structures may co-exist in the same cell at the same time. PMID- 19997844 TI - Increased cortical inhibition deficits in first-episode schizophrenia with comorbid cannabis abuse. AB - RATIONALE/OBJECTIVES: There is a high prevalence of substance use disorder (SUD) in first-episode schizophrenia (SZ), but its contribution to the underlying SZ pathophysiology remains unclear. Several studies using transcranial magnetic stimulation (TMS) have observed abnormalities in human motor cortex (M1) excitability in SZ. Studies on cortical excitability comparing SZ patients with and without comorbid substance abuse are lacking. METHODS: A total of 29 first episode SZ patients participated in this study; 12 had a history of comorbid cannabis abuse (SZ-SUD) and 17 did not (SZ-NSUD). We applied TMS to right and left M1 areas to assess the resting motor threshold (RMT), short-interval cortical inhibition (SICI), intracortical facilitation (ICF), and the contralateral cortical silent period (CSP). RESULTS: In SICI and ICF conditions, right M1 stimulation led to significantly higher motor evoked potential ratios in SZ-SUD compared to SZ-NSUD. This suggests lower cortical inhibition and increased ICF in first-episode SZ with previous cannabis abuse. There were no group differences in RMT and CSP duration. Neither were there any significant correlations between psychopathology (as indexed by Positive and Negative Syndrome Scale), disease characteristics, the extent of cannabis abuse, and TMS parameters (SICI, ICF, and CSP). CONCLUSIONS: Comorbid cannabis abuse may potentiate the reduced intracortical inhibition and enhanced ICF observed in first-episode SZ patients in some previous studies. This finding suggests an increased alteration of GABA(A) and NMDA receptor activity in cannabis-abusing first-episode patients as compared to schizophrenia patients with no history of substance abuse. This may constitute a distinct vulnerability factor in this special population. PMID- 19997847 TI - Cerebral metabolic changes in neurologically presymptomatic patients undergoing haemodialysis: in vivo proton MR spectroscopic findings. AB - OBJECTIVE: To prospectively investigate and detect early cerebral metabolic changes in patients with end-stage renal disease (ESRD) by using in vivo proton MR spectroscopy (MRS). METHODS: We enrolled 32 patients with ESRD and 32 healthy controls between the ages of 26 and 50 years. Short echo time single-voxel proton MRS was acquired from volumes of interest (VOIs) located in the frontal grey and white matter, temporal white matter and basal ganglia. The choline/phospatidylcholine (Cho), myo-inositol (mI), N-acetylaspartate (NAA) and total creatine (tCr) peaks were measured and the metabolic ratios with respect to tCr were calculated. RESULTS: In the ESRD group, significant elevations of the Cho/tCr and mI/tCr ratios were observed for the frontal grey matter, frontal white matter, temporal white matter and basal ganglia as compared with controls. There was no significant difference in the NAA/tCr ratios at all VOIs between the ESRD patients and the healthy controls. CONCLUSIONS: Proton MRS is a useful and non-invasive imaging tool for the detection of early cerebral metabolic changes in neurologically presymptomatic ESRD patients. PMID- 19997846 TI - Whole-body MRI using a sliding table and repositioning surface coil approach. AB - OBJECTIVE: To introduce and assess a new way of performing whole-body magnetic resonance imaging (MRI) using a non-integrated surface coil approach as available on most clinical MRI systems worldwide. METHODS: Ten consecutive asymptomatic subjects prospectively underwent whole-body MRI for health screening. Whole-body MRI included T1-, T2- and diffusion-weighted sequences, and was performed using a non-integrated surface coil to image four different stations without patient repositioning. The four separately acquired stations were merged, creating seamless coronal whole-body T1-, T2- and diffusion-weighted images. Anatomical alignment, image quality at the boundaries of adjacent stations, and overall image quality of all stations were qualitatively assessed. RESULTS: The average time (+/-SD) taken to change the surface coil from one station to the next station was 53.8 (+/-7.1) s. The average total extra examination time +/- SD was 2 min 41.4 s (+/-15.3 s). Anatomical alignment, image quality at the boundaries of adjacent stations, and overall image quality of all stations of T1-, T2- and diffusion-weighted whole-body MRI were overall graded as "good" to "excellent". CONCLUSION: This study shows that a time-efficient and high-quality whole-body MRI examination can easily be performed by using a non-integrated sliding surface coil approach. PMID- 19997848 TI - High-resolution computed tomography to differentiate chronic diffuse interstitial lung diseases with predominant ground-glass pattern using logical analysis of data. AB - OBJECTIVES: We evaluated the performance of high-resolution computed tomography (HRCT) to differentiate chronic diffuse interstitial lung diseases (CDILD) with predominant ground-glass pattern by using logical analysis of data (LAD). METHODS: A total of 162 patients were classified into seven categories: sarcoidosis (n = 38), connective tissue disease (n = 32), hypersensitivity pneumonitis (n = 18), drug-induced lung disease (n = 15), alveolar proteinosis (n = 12), idiopathic non-specific interstitial pneumonia (n = 10) and miscellaneous (n = 37). First, 40 CT attributes were investigated by the LAD to build up patterns characterising a category. From the association of patterns, LAD determined models specific to each CDILD. Second, data were recomputed by adding eight clinical attributes to the analysis. The 20 x 5 cross-folding method was used for validation. RESULTS: Models could be individualised for sarcoidosis, hypersensitivity pneumonitis, connective tissue disease and alveolar proteinosis. An additional model was individualised for drug-induced lung disease by adding clinical data. No model was demonstrated for idiopathic non-specific interstitial pneumonia and the miscellaneous category. The results showed that HRCT had a good sensitivity (>or=64%) and specificity (>or=78%) and a high negative predictive value (>or=93%) for diseases with a model. Higher sensitivity (>or=78%) and specificity (>or=89%) were achieved by adding clinical data. CONCLUSION: The diagnostic performance of HRCT is high and can be increased by adding clinical data. PMID- 19997849 TI - 64-Slice multidetector computed tomography evaluation of gastrointestinal tract perforation site: detectability of direct findings in upper and lower GI tract. AB - OBJECTIVE: To evaluate wall discontinuity, as observed using 64-slice multidetector-row computed tomography (64-MDCT), as a direct finding (DF) indicating the perforation site in patients with gastrointestinal (GI) tract perforations. METHODS: We retrospectively studied 41 consecutive patients presenting with acute abdomen and exhibiting extraluminal air (EA) on 64-MDCT. Three readers evaluated the distribution of EA, extraluminal faeces, dirty mass, dirty fat sign, extraluminal fluid collection and bowel wall thickening (i.e. conventional findings, CFs) as well as DFs. RESULTS: Twenty-two cases were surgically or endoscopically confirmed to have upper GI tract perforations, and 19 had lower GI tract perforations. The DFs correctly identified the sites of perforation in 80.5% of patients when 2-mm-thick imaging slices were used. For the detection of upper GI tract perforations, the sensitivity, specificity and accuracy were 95.5%, 94.7% and 95.1% for the DFs and 50.0%, 100% and 73.2% for the CFs, respectively. Significant differences in sensitivity (p < 0.001) and diagnostic accuracy (p < 0.05) were observed between the DFs and CFs for upper GI perforations but not for lower GI tract perforations. CONCLUSION: DFs of the perforation site by using 64-MDCT were more sensitive and accurate than CFs for the detection of upper GI tract perforations. PMID- 19997851 TI - DNA-based identification of forensically important Australian Sarcophagidae (Diptera). AB - The utility of the forensically important Sarcophagidae (Diptera) for time since death estimates has been severely limited, as morphological identification is difficult and thermobiological histories are inadequately documented. A molecular identification method involving the sequencing of a 658-bp 'barcode' fragment of the mitochondrial cytochrome oxidase subunit I (COI) gene from 85 specimens, representing 16 Australian species from varying populations, was evaluated. Nucleotide sequence divergences were calculated using the Kimura-two-parameter distance model and a neighbour-joining phylogenetic tree generated. All species were resolved as reciprocally monophyletic, except Sarcophaga dux. Intraspecific and interspecific variation ranged from 0.000% to 1.499% (SE = 0.044%) and 6.658% to 8.983% (SE = 0.653%), respectively. The COI 'barcode' sequence was found to be suitable for the molecular identification of the studied Australian Sarcophagidae: 96.5% of the examined specimens were assigned to the correct species. Given that the sarcophagid fauna is poorly described, it is feasible that the few incorrectly assigned specimens represent cryptic species. The results of this research will be instrumental for implementation of the Australian Sarcophagidae in forensic entomology. PMID- 19997850 TI - Alcohol-induced changes in the brain as assessed by MRI and CT. AB - This review provides an overview of structural magnetic resonance imaging and computed tomography findings of direct and indirect alcohol-related toxic effects on the brain. In addition to ethanol-related changes to the brain, this article will also describe imaging findings in the acute setting of methanol and ethylene glycol poisoning. Alcohol will lead to brain atrophy, osmotic myelinolysis, Marchiafava-Bignami disease and, especially when related to malnutrition, may also cause Wernicke encephalopathy. Brain atrophy can be reversible if alcohol abuse is stopped. If not treated, Wernicke encephalopathy can lead to coma and death and an early diagnosis is important for immediate initiation of thiamine substitution. As clinical symptoms are often unspecific, the radiologist plays an important role in the detection of alcohol abuse and its related clinical conditions. PMID- 19997852 TI - Altered myoplasmic Ca(2+) handling in rat fast-twitch skeletal muscle fibres during disuse atrophy. AB - Calcium-dependent signalling pathways are believed to play an important role in skeletal muscle atrophy, but whether intracellular Ca(2+) homeostasis is affected in that situation remains obscure. We show here that there is a 20% atrophy of the fast-type flexor digitorum brevis (FDB) muscle in rats hind limb unloaded (HU) for 2 weeks, with no change in fibre type distribution. In voltage-clamp experiments, the amplitude of the slow Ca(2+) current was found similar in fibres from control and HU animals. In fibres loaded with the Ca(2+) dye indo-1, the value for the rate of [Ca(2+)] decay after the end of 5-100-ms-long voltage-clamp depolarisations from -80 to +10 mV was found to be 30-50% lower in fibres from HU animals. This effect was consistent with a reduced contribution of both saturable and non-saturable components of myoplasmic Ca(2+) removal. However, there was no change in the relative amount of parvalbumin, and type 1 sarco-endoplasmic reticulum Ca(2+)-ATPase was increased by a factor of three in the atrophied muscles. Confocal imaging of mitochondrial membrane potential showed that atrophied FDB fibres had significantly depolarized mitochondria as compared to control fibres. Depolarization of mitochondria in control fibres with carbonyl cyanide-p-trifluoromethoxyphenylhydrazone induced a slowing of the decay of [Ca(2+)] transients accompanied by an increase in resting [Ca(2+)] and a reduction of the peak amplitude of the transients. Overall results provide the first functional evidence for severely altered intracellular Ca(2+) removal capabilities in atrophied fast-type muscle fibres and highlight the possible contribution of reduced mitochondrial polarisation. PMID- 19997853 TI - Coccoid Helicobacter pylori exists in the palatine tonsils of patients with IgA nephropathy. AB - PURPOSE: Helicobacter pylori infection is acquired by oral ingestion. H. pylori has been reported to be present in the palatine tonsils. To clarify the route and mode of infection, the prevalence of tonsillar H. pylori was evaluated, and an attempt was made to culture tonsillar H. pylori. METHODS: In a prospective study, 55 patients with recurrent pharyngotonsillitis or IgA nephropathy underwent a tonsillectomy. The carbon 13-urea breath test and enzyme-linked immunosorbent assay for the detection of H. pylori IgG antibodies in the serum were performed. Tonsillar H. pylori was cultured under conventional culture conditions for gastric H. pylori with or without the following shock methods; heat shock, hydrogen-peroxide-degrading compounds, or parasitizing amoebae. Immunofluorescence and immunoelectron microscopy using antibodies against H. pylori and cytotoxin-associated antigen A were used to identify tonsillar H. pylori. RESULTS: H. pylori in the coccoid form was present in tonsillar crypts. Of 55 patients, 43 (78.2%) had tonsillar H. pylori, and 15 (27.3%) were infected with gastric H. pylori. All patients with gastric H. pylori also had tonsillar H. pylori (p < 0.01). Cytotoxin-associated antigen A was observed in 38 (88.4%) of 43 tonsillar H. pylori. Tonsillar H. pylori could not be cultured in any culture conditions. All patients with IgA nephropathy had tonsillar H. pylori (p < 0.01). CONCLUSIONS: The present research might provide some insight into clarifying the route and mode of H. pylori infection. Our findings may indicate that tonsillar H. pylori is one of the antigens causative of IgAN. PMID- 19997854 TI - Prevalence of functional dyspepsia in an outpatient clinic with primary care physicians in Japan. AB - BACKGROUND: Since little is known about the prevalence of patients with functional gastrointestinal disorders (FGID), this study was performed to clarify the prevalence of FGID, especially functional dyspepsia (FD), in new patients of an outpatient clinic with primary care physicians in Japan. METHODS: We analyzed consecutive outpatients (n = 5183) who first visited the Department of General Medicine, Asahikawa Medical College Hospital, between April 2004 and March 2009. RESULTS: Out of 5813 patients, 818 (14.1%) visited because of abdominal symptoms. Final diagnoses of the 818 patients were FGID (n = 320, 39.1%), organic gastrointestinal diseases (n = 237, 28.9%), organic non-GI disease (n = 135, 16.5%), and others (n = 126, 15.4%). The 320 patients with FGID had FD (n = 170), irritable bowel syndrome (IBS) (n = 68), and other FGIDs (n = 88). The rate of FGID or FD in all patients was 5.5% or 2.9%, respectively. Among outpatients (n = 381) who complained of upper abdominal symptoms, approximately 45% had FD. There was no gender difference in the frequency of FGID, FD, or IBS in all ages of patients. A lower incidence of FD was shown in female patients over 70 years old and a higher incidence of IBS in male patients under 29 years old. DISCUSSION: These results suggest that the prevalence of FGID, especially FD, is very high in an outpatient clinic with primary care physicians in Japan. PMID- 19997855 TI - Serotonin transporter activity and serotonin concentration in platelets of patients with irritable bowel syndrome: effect of gender. AB - BACKGROUND: In the past decade, a strong argument has been built for the role of serotonin (5HT) and the serotonin transporter (SERT) in irritable bowel syndrome (IBS). However, it is still not clear how SERT contributes to this clinically heterogeneous disease. The present study addressed this issue by implementing platelet (plt) markers of SERT activity in the assessment protocol. METHODS: Fasting blood samples of 149 (51 male/98 female) subjects with Rome II and III defined IBS subtypes, and 163 healthy control subjects (CSs; 75 male/88 female) were analyzed for platelet 5HT concentration and 5HT uptake activity [maximum uptake rate (V (max)) and affinity constant (K (m))]. RESULTS: Gender had a significant impact on platelet markers of SERT activity. Male IBS patients showed significantly lower median V (max) and K (m) values than the male CS (V (max) 1.706 vs. 2.148 nmol/10(9) plts x min, P < 0.001; K (m) 346 vs. 410 nmol, P = 0.008) without any significant reduction in platelet 5HT concentration (362 vs. 394 ng/10(9) plts). On the other hand, V (max) values were not different between female IBS patients and female CS (1.642 vs. 1.741 nmol/10(9) plts x min), but platelet 5HT concentration was significantly lower in females with diarrhea predominant IBS (363 vs. 435 ng/10(9) plts, P < 0.05). CONCLUSION: Although an absolute extrapolation from platelets to the gastrointestinal tissue does not appear to be justified, our findings demonstrated that the contribution of disturbed SERT activity to IBS is not uniform and is possibly gender-specific. The results suggest that an assessment of SERT function in platelets may help to elucidate the differences between IBS patients in response to drugs affecting the 5HT system. PMID- 19997856 TI - Molecular signatures of noncancerous liver tissue can predict the risk for late recurrence of hepatocellular carcinoma. AB - Hepatocellular carcinoma (HCC) is an aggressive malignancy mainly due to tumor metastases or recurrence even after undergoing potentially curative treatment. There are two types of HCC recurrence. The early and late tumor recurrences appear in distinct biological contexts, and their clinical courses are quite different. Therefore, it is important to precisely and distinctly discriminate the risk of each type of HCC recurrence. Many researchers have used DNA microarray technology to reclassify HCC with respect to its malignant potential. Some of these studies successfully identified specific gene-expression signatures derived from the cancerous tissues of HCC for predicting the early recurrence due to intrahepatic metastasis. However, there are no well-defined predictors for late recurrence. Recently, a few studies have focused on the nontumorous portion of liver tissues to predict late recurrence, possibly due to de novo hepatocarcinogenesis based on the idea of "field cancerization." This study reviewed the possible value of a gene-expression analysis of noncancerous liver tissue to clarify the risk for multicentric late recurrence of HCC. These findings may have important implications for chemopreventive strategies and tailored surveillance programs. Furthermore, this approach may also be applicable to other multifocal tumors, such as head and neck carcinoma. PMID- 19997857 TI - -651C/T promoter polymorphism in the CD14 gene is associated with severity of acute pancreatitis in Japan. AB - PURPOSE: This study aimed to clarify the association of the promoter variants in the CD14 gene with pancreatic diseases in Japan. METHODS: Three hundred forty-six unrelated patients with acute pancreatitis (AP) (107 with severe and 239 with mild), 263 patients with chronic pancreatitis (CP), 264 patients with pancreatic neoplasm, and 319 healthy controls were genotyped for the single nucleotide polymorphisms at positions -260 and -651 from the AUG start codon in the CD14 gene by polymerase chain reaction-restriction enzyme digestion. RESULTS: The allele and genotype frequencies of the -260C/T and -651C/T polymorphisms did not differ between controls and patients with AP. In subgroup analyses, patients with severe AP had more -651C allele than controls [P = 0.005; odds ratio (OR) 1.71; 95% confidence interval (CI) = 1.18-2.49] or patients with mild AP (P = 0.001; OR 1.95; 95% CI = 1.33-2.85). Genotype -651CC was more common (P = 0.001 vs. controls and P = 0.001 vs. mild AP), and -651CT was less (P = 0.009 vs. controls and P = 0.007 vs. mild AP) in patients with severe AP than in healthy controls or patients with mild AP. The frequencies of pseudocyst development and requirement of surgery were higher in AP patients with -651CC than in those without this genotype. The -260C/T polymorphism was not associated with the severity of AP. The allele and genotype frequencies of both polymorphisms did not differ between controls and patients with CP or pancreatic neoplasm. CONCLUSION: -651C/T promoter polymorphism in the CD14 gene was associated with severity of AP in Japan. PMID- 19997858 TI - Association of a functional variant of neuronal nitric oxide synthase gene with self-reported impulsiveness, venturesomeness and empathy in male offenders. AB - It has been shown that a functional promoter dinucleotide repeat length variation of the neuronal nitric oxide synthase gene (NOS1 Ex1f-VNTR) is associated with impulsivity-related behavioral phenotypes. In this study, the Eysenck Impulsivity Questionnaire (IVE-7) was administered to 182 male offenders to prove the hypothesis that NOS1 Ex1f-VNTR is associated with self-reported impulsiveness, venturesomeness and empathy. Multivariate analysis of variance (MANCOVA) revealed a significant multivariate effect of NOS1 Ex1f-VNTR genotype on IVE-7 measures (P = 0.0006). The effect was more pronounced regarding impulsiveness and empathy (P = 0.0052 and P = 0.0036, respectively) as compared with venturesomeness, which was only of borderline significance. The findings give additional evidence that NOS-I is involved in the regulation of impulsive personality traits and support the notion that the NOS1 gene takes part in the regulation of social behavior. PMID- 19997860 TI - Expression and function of histamine H4 receptor in mouse splenic dendritic cells. PMID- 19997859 TI - Two-year clinical performance of glass ionomer and resin composite restorations in xerostomic head- and neck-irradiated cancer patients. AB - The aim of this study was to evaluate the clinical performance of adhesive filling materials in class V cavities in xerostomic head- and neck-irradiated cancer patients, in terms of marginal adaptation, anatomical form and recurrent caries. We selected 35 high-caries-risk, post-radiation, xerostomic adults with >=3 cervical carious lesions in the same arch. Every patient received a KetacFil (KF), PhotacFil (PF) and Herculite XRV (HX) restoration. Patients were instructed to use a neutral 1% sodium fluoride gel in custom trays, on a daily basis. After 6, 12, 18 and 24 months, the restorations were examined for material loss, marginal integrity and recurrent caries. Fluoride compliance was determined at each recall appointment and recorded as the percentage of recommended use during that interval [compliance of <=50% = NFUs, >50% = FUs]. Only 30 patients were available for recall at 6 months, with 28 patients at 12 and 18 months, and 27 patients at 24 months. In the NFU group, differences in recurrent caries were found between KF and HX at all observation times (p < 0.05). Differences (p < 0.05) in adaptation and/or anatomical form were found between KF and PF in NFUs after 18 and 24 months. In FUs, significant differences were observed between KF and PF, and KF and HX after 6 and 12 months, between KF and HX, PF and HX after 18 and 24 months. In summary, glass ionomers (especially the conventionally setting formulation) provide clinical caries inhibition but erode easily, while composite resin provides greater structural integrity. PMID- 19997861 TI - Efficacy, pharmacokinetics, safety, and tolerability of Flebogamma 10% DIF, a high-purity human intravenous immunoglobulin, in primary immunodeficiency. AB - BACKGROUND: Flebogamma 10% DIF represents an evolution of intravenous immune globulin from the previous 5% product to be administered at higher rates and with smaller infusion volumes. Pathogen safety is enhanced by the combination of multiple methods with different mechanisms of action. OBJECTIVE: The objective of this study as to evaluate the efficacy, pharmacokinetics, and safety of Flebogamma 10% DIF for immunoglobulin replacement therapy in primary immunodeficiency diseases (PIDD). METHODS: Flebogamma 10% DIF was administered to 46 subjects with well-defined PIDD at a dose of 300-600 mg/kg every 21-28 days for 12 months. RESULTS: Serious bacterial infection rate was 0.025/subject/year. Half-life in serum of the administered IgG was approximately 35 days. No serious treatment-related adverse event (AE) occurred in any patient. Most of the potentially treatment-related AEs occurred during the infusion, accounting for 20% of the 601 infusions administered. CONCLUSIONS: Flebogamma 10% DIF is efficacious and safe, has adequate pharmacokinetic properties, and is well tolerated for the treatment of PIDD. PMID- 19997862 TI - HIV and related risk behavior among men who have sex with men in Zanzibar, Tanzania: results of a behavioral surveillance survey. AB - We conducted a respondent driven sampling survey to estimate HIV prevalence and risk behavior among men who have sex with men (MSM) in Unguja, Zanzibar. Men aged >= 15 years living in Unguja and reporting anal sex with another man in the past 3 months were asked to complete a questionnaire and provide specimens for biologic testing. HIV prevalence was 12.3% (95% confidence interval 8.7, 16.3). HIV infection was associated with injecting drugs in the past 3 months, Hepatitis C virus infection and being paid for sex in the past year. Interventions for MSM in Zanzibar are needed and should include linkages to prevention, care and treatment services. PMID- 19997863 TI - Isolation and identification of mucinolytic actinomycetes. AB - Biochemical and physiological tests, and 16S rRNA gene sequences, were used to classify nine Actinomycete strains isolated from soil and sand samples in Thailand. These strains were isolated based on their ability to readily degrade mucin glycoproteins. A turbidometric based mucinolytic assay was developed to confirm this. In addition all strains showed significant production of proteases. Phylogenetic analysis of the strains revealed that from the nine isolated Actinomycete strains eight were closely related to Streptomyces species and one was identified as belonging to the genus Kitasatospora. The biochemical and physiological tests performed identified two strain pairs that were similar (with only 3.9% difference observed) and this was in accordance with the phylogenetic results obtained. The remaining strains were distinct from each other, with the soil-isolated strains forming a separate clade to the sand-isolated strains in the inferred phylogenetic trees. The isolated mucinolytic Actinomycete strains will be the subject of further investigations into their proteolytic and glycosidic activity. Mucin degrading enzymes such as these are studied for their potential to be used for the development of a drug delivery system. PMID- 19997864 TI - Transferability of a tetracycline resistance gene from probiotic Lactobacillus reuteri to bacteria in the gastrointestinal tract of humans. AB - The potential of Lactobacillus reuteri as a donor of antibiotic resistance genes in the human gut was investigated by studying the transferability of the tetracycline resistance gene tet(W) to faecal enterococci, bifidobacteria and lactobacilli. In a double-blind clinical study, seven subjects consumed L. reuteri ATCC 55730 harbouring a plasmid-encoded tet(W) gene (tet(W)-reuteri) and an equal number of subjects consumed L. reuteri DSM 17938 derived from the ATCC 55730 strain by the removal of two plasmids, one of which contained the tet(W) gene. Faecal samples were collected before, during and after ingestion of 5 x 10(8) CFU of L. reuteri per day for 14 days. Both L. reuteri strains were detectable at similar levels in faeces after 14 days of intake but neither was detected after a two-week wash-out period. After enrichment and isolation of tetracycline resistant enterococci, bifidobacteria and lactobacilli from each faecal sample, DNA was extracted and analysed for presence of tet(W)-reuteri using a real-time PCR allelic discrimination method developed in this study. No tet(W)-reuteri signal was produced from any of the DNA samples and thus gene transfer to enterococci, bifidobacteria and lactobacilli during intestinal passage of the probiotic strain was non-detectable under the conditions tested, although transfer at low frequencies or to the remaining faecal bacterial population cannot be excluded. PMID- 19997865 TI - Neighborhood factors affecting rates of sexually transmitted diseases in Chicago. AB - High rates of gonorrhea have been shown to be associated with high rates of incarceration in the prior year. One hypothesized chain of events is that there is a negative effect of incarceration on neighborhood social characteristics, which in turn affect behaviors facilitating transmission of sexually transmitted diseases (STDs). This study examined whether neighborhood characteristics were associated with the incidence of STDs and homicide rates as a proxy for incarceration rates. Data were from the 1995 Program on Human Development in Chicago Neighborhoods, the Chicago Health Department, and the Chicago Police Department. Neighborhood gonorrhea rates increased by 192.2 (95% confidence interval (CI) 131.6, 252.9) cases per 100,000 population with a change from the 25th to the 75th percentile of social disorder. This rate difference was a value greater than the median neighborhood gonorrhea rate. Similar increases were observed for other neighborhood measures and for Chlamydia infection. We hypothesize that high rates of incarceration may play a role in undermining neighborhood social cohesion and control. Using homicide rates as a proxy for incarceration, a change from the 25th to the 75th percentile of 1995 neighborhood homicide rates yielded a gonorrhea rate increase of 164.6 (95% CI 124.4, 204.7) cases per 100,000. Factors that undermine the social fabric of a community can become manifest in health outcomes such as STDs. The effects of high rates of incarceration on neighborhoods merit further exploration. PMID- 19997866 TI - Entry of bovine viral diarrhea virus into ovine cells occurs through clathrin dependent endocytosis and low pH-dependent fusion. AB - Although mechanisms of bovine viral diarrhea virus (BVDV) entry into bovine cells have been elucidated, little is known concerning pestivirus entry and receptor usage in ovine cells. In this study, we determined the entry mechanisms of BVDV-1 and BVDV-2 in sheep fetal thymus cells. Both BVDV-1 and BVDV-2 infections were inhibited completely by chlorpromazine, beta-methyl cyclodextrin, sucrose, bafilomycin A1, chloroquine, and ammonium chloride. Simultaneous presence of reducing agent and low pH resulted in marked loss of BVDV infectivity. Moreover, BVDV was unable to fuse with ovine cell membrane by the presence of reducing agent or low pH alone, while combination of both led to fusion at low efficiency. Furthermore, sheep fetal thymus cells acutely infected with BVDV-1 or BVDV-2 were found protected from heterologous BVDV infection. Taken together, our results showed for the first time that entry of both BVDV-1 and BVDV-2 into ovine cells occurred through clathrin-dependent endocytosis, endosomal acidification, and low pH-dependent fusion following an activation step, besides suggesting the involvement of a common ovine cellular receptor during attachment and entry. PMID- 19997867 TI - Dexamethasone protects cultured rat hepatocytes against cadmium toxicity: involvement of cellular thiols. AB - In the present study, the effects of dexamethasone on cadmium-induced toxicity were evaluated in isolated rat hepatocytes. Hepatocytes were cultured for 24 h in William's E medium containing fetal calf serum (10%), insulin (0.1 IU/ml), and glucagon (0.01 microM) in the absence or presence of 0.1 microM dexamethasone. Cadmium chloride, 5 or 10 microM, was added to the medium and the toxicity was evaluated for up to 48 h after treatment. Lactate dehydrogenase (LDH) release, the reduced and oxidized glutathione ratio (GSH/GSSG), protein-SH groups, and lipid peroxidation levels were evaluated. Cadmium induced a dose- and time dependent LDH release in control hepatocytes at 24 h (Cd 10 microM 42%) while hepatocytes pretreated with dexamethasone showed lower necrosis (Cd 10 microM 12% at 24 h). GSH/GSSH ratio and protein-SH groups were higher while lipid peroxidation was lower in dexamethasone-treated hepatocytes as compared with untreated cells. In conclusion, cadmium toxicity was associated with an increase in intracellular oxidative stress responsible for accelerated cell death. The use of dexamethasone prevented cadmium damage, suggesting that the cytoprotective action of this hormone is related to its effect in preventing changes in thiols such as glutathione and protein-SH groups. PMID- 19997868 TI - Development of a scaffoldless three-dimensional engineered nerve using a nerve fibroblast co-culture. AB - Nerve grafts are often required to replace tissue damaged by disease, surgery, or extensive trauma. Limitations such as graft availability, donor site morbidity, and immune rejection have led investigators to develop strategies to engineer nerve tissue. The goal of this study was to fabricate a scaffoldless three dimensional (3D) nerve construct using a co-culture of fetal nerve cells with a fibroblast monolayer and allow the co-culture to remodel into a 3D construct with an external fibroblast layer and an internal core of interconnected neuronal cells. Primary fibroblasts were seeded on laminin-coated plates and allowed to form a confluent monolayer. Neural cells isolated from E-15 spinal cords were seeded on top of the fibroblast monolayer and allowed to form a networked monolayer across the monolayer of fibroblasts. Media shifts initiated contraction of the fibroblast monolayer and a remodeling of the co-culture into a 3D construct held statically in place by the two constraint pins. Immunohistochemistry using S100 (Schwann cell), beta3-tubulin, DAPI, and collagen I indicated an inner core of nerve cells surrounded by an external layer of fibroblasts. Conduction velocities of the 3D nerve and control (fibroblast-only) constructs were measured in vitro and compared to in vivo measures of neonatal sciatic nerve. The conduction velocities of the nerve constructs were comparable to 24-d-old neonatal nerve. The presence of Schwann cells and the ability to conduct neuronal signals in vitro suggest the scaffoldless 3D nerve constructs will be a viable option for nerve repair. PMID- 19997869 TI - A protocol to effectively create single cell suspensions of adherent cells for multiparameter high-throughput flow cytometry. AB - High-throughput flow cytometry of adherent cells is difficult because the creation of single cell suspensions can damage cells and yield artificial results. We describe a protocol to increase the single cell suspension yield of adherent human cells without injury. Doxorubicin, a cytotoxic agent, was administered to adherent human pancreatic carcinoma cell lines (Panc-1 and AsPC 1) to produce alterations in the cell cycle and intracellular protein expression. The cells in 96-well plates were disassociated using a collagenase and trypsin mixture. Fluorescence-activated high-throughput flow cytometry evaluated cellular viability as well as surface and intracellular protein expression. Cell cycle analysis was performed using 7-aminoactinomycin D and intracellular protein characterization was performed using a fluorescein-labeled monoclonal antibody against activated caspase-3. The collagenase-trypsin-based protocol increased single cell events from 31.9 +/- 0.5% using trypsin alone (standard) to a range of 62.1% to 85.5% without adversely affecting viability. High-throughput flow cytometry demonstrated that the addition of collagenase to the disassociation solution not only permitted significantly higher rates of single cell creation, but it did not negatively affect the doxorubicin-induced protein expression. This protocol allows for expedient and effective disassociation of adherent human cells in order to investigate alterations in specific cellular enzymes and pathways. PMID- 19997870 TI - Establishment and characterization of androgen-independent human prostate cancer cell lines, PcBra1, PcBra2, and PcBra3. AB - One of the main obstacles for understanding biological events involved in cancer is the lack of experimental models for in vitro studies especially for prostate cancer (PC). There are a limited number of PC cell lines being the majority originated from metastatic tumors mostly acquired from American Tissue Cell Culture which demands importation an expensive and bureaucratic process. Also it is well known that there are ethnic differences between populations concerning the behavior of tumors and the research based on cell lines derived from Brazilians should be interesting. Our aim was to develop tumor cell lines from primary PC. PMID- 19997871 TI - Humanin and the receptors for humanin. AB - Alzheimer's disease (AD) is a prevalent dementia-causing neurodegenerative disease. Neuronal death is closely linked to the progression of AD-associated dementia. Accumulating evidence has established that a 24-amino-acid bioactive peptide, Humanin, protects neurons from AD-related neuronal death. A series of studies using various murine AD models including familial AD gene-expressing transgenic mice have shown that Humanin is effective against AD-related neuronal dysfunction in vivo. Most recently, it has been shown that Humanin inhibits neuronal cell death and dysfunction by binding to a novel IL-6-receptor-related receptor(s) on the cell surface involving CNTFRalpha, WSX-1, and gp130. These findings suggest that endogenous Humanin [or a Humanin-like substance(s)] may suppress the onset of AD-related dementia by inhibiting both AD-related neuronal cell death and dysfunction. PMID- 19997872 TI - Immunohistological profile of the Ras homologous B protein (RhoB) in human testes showing normal spermatogenesis, spermatogenic arrest and Sertoli cell only syndrome. AB - Ras homologous B protein (RhoB) belongs to the Ras homologous subfamily which consists of low molecular weight (21 kDa) GTP-binding proteins. Rho proteins are regulatory molecules associated with various kinases and as such they mediate changes in cell shape, contractility, motility and gene expression. To date, no data are available about the expression pattern of RhoB protein in the human testis showing normal and abnormal spermatogenesis. The present study addresses these issues. Human testicular biopsy specimens were obtained from patients suffering from post-testicular infertility (testis showing normal spermatogenesis, 10 cases) and testicular infertility (testis showing Sertoli cell only syndrome and spermatogenic arrest, 10 patients each). The expression of RhoB was examined using in situ immunofluorescent staining methods. In testes showing normal spermatogenesis, RhoB had a strong expression in the seminiferous epithelium (cytoplasm of Sertoli-cells, spermatogonia and spermatocytes) and in the interstitium (Leydig cells). RhoB expression was weak in the myofibroblasts and absent in the spermatids and sperms. In the testes showing abnormal spermatogenesis, RhoB expression was moderate in the seminiferous epithelium (cytoplasm of Sertoli cells, spermatogonia and spermatocytes) and was completely absent in the Leydig cells, myofibroblasts, spermatids and sperms. To the best of our knowledge, this study provides the first morphological indication that RhoB protein is expressed in human testis and its expression undergoes testicular infertility associated changes. These findings suggest the involvement of RhoB in the process of spermatogenesis in human and their possible therapeutic ramifications in testicular infertility are open for further investigations. PMID- 19997873 TI - Glutamate promotes cell growth by EGFR signaling on U-87MG human glioblastoma cell line. AB - Accumulating evidences suggest that glutamate plays a key role in the proliferation and invasion of malignant glioblastoma (GBM) tumors. It has been shown that GBM cells release and exploit glutamate for proliferation and invasion through AMPA glutamate receptors. Additionally, amplification of the epidermal growth factor receptor (EGFR) gene occurs in 40-50% of GBM. Since, PI3K/Akt is considered one of the main intracellular pathways involved in EGFR activation, AKT functions could trigger EGFR signaling. Thus, we investigated whether EGFR phospho-Akt pathway is involved on the glutamate inducing U-87MG human GBM cell line proliferation. For these purpose, we treated the U-87MG cell line with 5 to 200 mM of glutamate and assessed the number of viable cells by trypan blue dye exclusion test. An increase in cell number (50%) was found at 5 mM glutamate, while the addition of DNQX (500 microM), an antagonist of AMPA receptor, inhibited the effect of glutamate on the U87-MG cells proliferation. Also, at 5 mM glutamate we observed an increase on the EGFR and phospho-Akt contents evaluated by immunohistochemistry. Moreover, U-87MG cells treated with glutamate exhibited an increase about 2 times in the EGFR mRNA expression. While, in the presence of the anti-EGFR gefitinib (50 muM) or the PI3K inhibitor wortmannin (5 muM), the U-87MG proliferation was restored to control levels. Together, our data suggest that glutamate signaling mediated by AMPA receptor induces U-87MG human GBM cell line proliferation via EGFR-phospho-Akt pathway. PMID- 19997875 TI - Thermal melting studies of ligand DNA interactions. AB - A simple thermal melting experiment may be used to demonstrate the stabilization of a given structure by a ligand (usually a small molecule, sometimes a peptide). Preparation of the sample is straightforward, and the experiment itself requires an inexpensive apparatus. Furthermore, reasonably low amounts of sample are required. A qualitative analysis of the data is simple: An increase in the melting temperature (T(m)) indicates preferential binding to the folded form as compared to the unfolded form. However, it is perilous to derive an affinity constant from an increase in T(m) as other factors play a role. PMID- 19997874 TI - Quantitative analysis of small molecule-nucleic acid interactions with a biosensor surface and surface plasmon resonance detection. AB - Surface plasmon resonance (SPR) technology with biosensor surfaces has become a widely-used tool for the study of nucleic acid interactions without any labeling requirements. The method provides simultaneous kinetic and equilibrium characterization of the interactions of biomolecules as well as small molecule biopolymer binding. SPR monitors molecular interactions in real time and provides significant advantages over optical or calorimetic methods for systems with strong binding coupled to small spectroscopic signals and/or reaction heats. A detailed and practical guide for nucleic acid interaction analysis using SPR biosensor methods is presented. Details of the SPR technology and basic fundamentals are described with recommendations on the preparation of the SPR instrument, sensor chips, and samples, as well as extensive information on experimental design, quantitative and qualitative data analysis and presentation. A specific example of the interaction of a minor-groove-binding agent with DNA is evaluated by both kinetic and steady-state SPR methods to illustrate the technique. Since the molecules that bind cooperatively to specific DNA sequences are attractive for many applications, a cooperative small molecule-DNA interaction is also presented. PMID- 19997876 TI - Circular and linear dichroism of drug-DNA systems. AB - When a drug binds to DNA, its electronic structure is perturbed, and it perturbs the DNA's electronic structure. The resulting change to the electronic spectroscopy can be used to probe the drug-DNA interaction. This chapter outlines how circular and linear dichroism spectroscopy can be used to provide information about drug-DNA systems. Circular dichroism spectroscopy involves measuring the difference in absorption of left and right circularly polarized light. It is uniquely sensitive to the helicity of the molecules being studied. Linear dichroism, as the name implies, involves measuring the difference in absorption of light linearly polarized parallel and perpendicular to an orientation axis. Linear dichroism provides information about the relative orientations of subunits of an interacting system. The material presented in this chapter is by no means comprehensive; the aim is to enable the user to collect reasonable quality data and to interpret it. PMID- 19997878 TI - Quantification of binding data using capillary electrophoresis. AB - The design of new DNA-targeted molecules, primarily for use in the therapy of diseases such as cancer, relies on the assessment of both affinity for DNA and selectivity of binding to chosen base pair sequences. Capillary electrophoresis, with a polymer added to the running buffer, is very well suited to the separation of oligonucleotides in the range 12-20 base pairs, with the separation based on length rather than base pair sequence. In this way, it is possible to conduct competition experiments using mixtures of up to four oligonucleotides and giving a direct measure of the relative affinity of high-affinity ligands, specifically those binding in the minor groove with slow on-off rates. The relative affinities can be securely quantified, even where the affinities are very high. Working from first principles, it is shown that the measurement of absolute affinities presents various problems, not least that the concentration of DNA and ligand used in the experiment will affect the magnitude of K(d), which is not constant. PMID- 19997877 TI - Drug binding to DNA x RNA hybrid structures. AB - The DNA x RNA hybrid duplexes are functionally important structures in gene expression that are underutilized as potential drug targets. Several tools are described here for the discovery and characterization of small molecules capable of the selective recognition of DNA x RNA hybrid structures. Competition dialysis and thermal denaturation of mixtures of polynucleotide structures can be used to identify small molecules that bind selectively to DNA x RNA hybrids. An assay that measures small molecule inhibition of RNase H can be used to measure a functional response to these ligands. PMID- 19997879 TI - Determination of equilibrium association constants of ligand-DNA complexes by electrospray mass spectrometry. AB - Electrospray mass spectrometry can be used to detect ligand-DNA noncovalent complexes formed in solution. This chapter describes how to determine equilibrium association constants of the complexes. Particular attention is devoted to describing how to tune an electrospray mass spectrometer using a 12-mer oligodeoxynucleotides duplex in order to perform these experiments. This protocol can then be applied to any nucleic acid structure that can be ionized with electrospray mass spectrometry. PMID- 19997880 TI - Detection of adriamycin-DNA adducts by accelerator mass spectrometry. AB - There have been many attempts in the past to determine whether significant levels of Adriamycin-DNA adducts form in cells and contribute to the anticancer activity of this agent. Supraclincal drug levels have been required to study drug-DNA adducts because of the lack of sensitivity associated with many of the techniques employed, including liquid scintillation counting of radiolabeled drug. The use of accelerator mass spectrometry (AMS) has provided the first direct evidence of Adriamycin-DNA adduct formation in cells at clinically relevant Adriamycin concentrations. The exceedingly sensitive nature of AMS has enabled over three orders of magnitude increased sensitivity of Adriamycin-DNA adduct detection (compared to liquid scintillation counting) and has revealed adduct formation within an hour of drug treatment. The rigorous protocol required for this approach, together with many notes on the precautions and procedures required in order to ensure that absolute levels of Adriamycin-DNA adducts can be determined with good reproducibility, is outlined in this chapter. PMID- 19997881 TI - Molecular modelling methods to quantitate drug-DNA interactions. AB - We describe a molecular modelling method for calculating the binding affinity of ligands for DNA. Though theoretically applicable to any form of noncovalent interaction, we concentrate on the case of predicting the sequence selectivity of a minor-groove binding ligand. The method is based on performing molecular dynamics (MD) simulations on DNA sequences, with and without the ligand bound, and postprocessing the molecular dynamics trajectory data to obtain approximate free energies of binding. We discuss issues relating to the preparation of the structures for simulation, choices for the molecular dynamics simulation method itself, methods for evaluating the reliability and stability of the simulation data, and finally alternative approaches to postprocessing the data to extract approximate free energies of binding. PMID- 19997882 TI - Application of anomalous diffraction methods to the study of DNA and DNA complexes. AB - Anomalous scattering is commonly used to solve X-ray structures. As discussed here, anomalous scattering is also useful for characterizing complex systems with mixed and partial occupancies, where true electron density is represented by unresolvable ensemble averages. The solvent environment surrounding nucleic acids is an example of such a system, as are some DNA-ligand systems. The atomic number and wavelength dependencies of anomalous scattering allow one to filter out the electron densities of C, N, and O, and to cleanly visualize the electron densities of heavier atoms. Therefore, anomalous scattering can make beacons of selected atoms. In addition, anomalous scattering provides a model-independent method for determining atomic identities. Here, we describe applications of anomalous scattering to the structure determination of DNA-platinum complexes and in cation associations of free DNA, of DNA-anthracycline complexes, of chemically modified DNA, and of DNA-protein complexes. The utility of Rb(+) and Tl(+) as K(+) substitutes is supported by similarities in Rb(+) and Tl(+) association with DNA. PMID- 19997883 TI - DNase I footprinting. AB - Footprinting is a method for determining the sequence selectivity of DNA-binding compounds in which ligands protect DNA from cleavage at their binding sites. Footprinting templates are typically 50-200 base pairs long, and DNase I is the most commonly used nuclease for these experiments. This chapter describes the preparation and labelling of suitable DNA footprinting substrates, the footprinting experiment itself, and the way in which these data can be used to estimate the dissociation constant of the interaction. PMID- 19997884 TI - Methods to characterize the effect of DNA-modifying compounds on nucleosomal DNA. AB - Eukaryotic DNA forms a complex with an equal mass of proteins to form chromatin. To fully understand the action of DNA-reactive antitumor antibiotics in the cell, their effect must be studied in a chromatin context. In particular, it is of interest to investigate how the distortion of DNA, in the context of a nucleosome, affects the action of drugs with either monoalkylation or crosslinking activity, and how modified DNA is assembled into chromatin. Here, we present experimental approaches that allow one to compare the effect of such drugs on free DNA and nucleosomes. We find significant differences that likely arise from the different geometry of nucleosomal DNA compared to free DNA and also find that drug-mediated DNA crosslinking affects nucleosome assembly. PMID- 19997885 TI - REPSA: combinatorial approach for identifying preferred drug-DNA binding sequences. AB - Many DNA-binding small molecules, typically those with a molecular mass less than 1,000 g/mol, recognize duplex DNA with some degree of sequence specificity. These include drugs used to treat several human diseases, including viral and bacterial infections, malaria, and cancer. Determining the binding specificity of DNA binding molecules can be important for their development, especially if they are being designed to target specific DNA sequences. A limited amount of information can be obtained through the study of small molecule binding to defined naturally occurring or synthetic DNA sequences; however, a full picture of a small molecule's binding specificity can only be obtained through combinatorial means, whereby vast libraries of sequences are screened. Several combinatorial methods have been developed for the study of ligand-DNA interactions, but only one method, Restriction Endonuclease Protection Selection and Amplification (REPSA), is generally applicable to the study of native small molecule-DNA complexes under physiologic conditions. REPSA may be used with both covalent and noncovalent small molecule-DNA complexes and with mixtures of small molecules with relatively unknown identities and properties. Thus, REPSA is a powerful, versatile, general method for the combinatorial determination of small molecule-DNA binding specificity and a functional means for drug discovery and characterization. PMID- 19997886 TI - In vitro transcription assay for resolution of drug-DNA interactions at defined DNA sequences. AB - A major class of anticancer agents in current clinical use exerts its anticancer effects by binding covalently or non-covalently to DNA. A detailed understanding of the nature of these drug-DNA complexes would be expected to lead to better uses of these drugs, and also assist with the design of improved drug derivatives. Here, we present a transcriptional footprinting assay that can be implemented to define the DNA sequence specificity and kinetics associated with drug-DNA complexes. The basic steps involve the formation of drug-DNA complexes, the formation of synchronised initiated transcripts, and finally transcriptional elongation to reveal drug blockage sites that impede the progression of RNA polymerase. We have used the "in vitro transcription assay" to investigate many covalent drug-DNA interactions; most notably those obtained using anthracycline anticancer agents such as doxorubicin and anthracenedione-based anticancer agents, including mitoxantrone and pixantrone. PMID- 19997887 TI - In vitro footprinting of promoter regions within supercoiled plasmid DNA. AB - Polypurine/polypyrimidine (pPu/pPy) tracts, which exist in the promoter regions of many growth-related genes, have been proposed to be very dynamic in their conformation. In this chapter, we describe a detailed protocol for DNase I and S1 nuclease footprinting experiments with supercoiled plasmid DNA containing the promoter regions to probe whether there are conformational transitions to B-type DNA, melted DNA, and G-quadruplex structures within this tract. This is demonstrated with the proximal promoter region of the human vascular endothelial growth factor (VEGF) gene, which also contains multiple binding sites for Sp1 and Egr-1 transcription factors. PMID- 19997888 TI - Topoisomerase I-mediated DNA relaxation as a tool to study intercalation of small molecules into supercoiled DNA. AB - Several biochemical and biophysical methods are available to study the intercalation of a small molecule between two consecutive base pairs of DNA. Among them, the topoisomerase I-mediated DNA relaxation assay has proved highly efficient, relatively easy to handle and very informative to investigate drug binding to DNA. The test relies on the use of a supercoiled plasmid to mimic the topological constraints of genomic DNA. The three main components of the assay - the topoisomerase I enzyme, DNA helix and intercalating small molecules - are presented here in a structural context. The principle of the assay is described in detail, along with a typical experimental protocol. PMID- 19997889 TI - A high-throughput assay for DNA topoisomerases and other enzymes, based on DNA triplex formation. AB - We have developed a rapid, high-throughput assay for measuring the catalytic activity (DNA supercoiling or relaxation) of topoisomerase enzymes that is also capable of monitoring the activity of other enzymes that alter the topology of DNA. The assay utilises intermolecular triplex formation to resolve supercoiled and relaxed forms of DNA, the principle being the greater efficiency of a negatively supercoiled plasmid to form an intermolecular triplex with an immobilised oligonucleotide than the relaxed form. The assay provides a number of advantages over the standard gel-based methods, including greater speed of analysis, reduced sample handling, better quantitation and improved reliability and accuracy of output data. The assay is performed in microtitre plates and can be adapted to high-throughput screening of libraries of potential inhibitors of topoisomerases including bacterial DNA gyrase. PMID- 19997890 TI - Measurement of DNA interstrand crosslinking in individual cells using the Single Cell Gel Electrophoresis (Comet) assay. AB - The Single Cell Gel Electrophoresis (Comet) assay, originally developed to allow visualisation of DNA strand break damage in individual cells, has been adapted to measure DNA interstrand cross-links. DNA interstrand cross-links are formed in cells by a number of commonly used cancer chemotherapy agents and are considered to be the critical lesion formed by such agents. This technique allows the analysis of DNA interstrand cross-link formation and repair at a single cell level, requires few cells, allows the determination of heterogeneity of response within a cell population and is sensitive enough to measure DNA interstrand cross links at pharmacologically relevant doses. The method can be applied to any in vitro or in vivo application where a single cell suspension can be obtained. The method has also become invaluable in studies using human tissue and can be used as a method for pharmacodynamic analysis in early clinical trials. PMID- 19997891 TI - Measurement of DNA interstrand crosslinking in naked DNA using gel-based methods. AB - Bifunctional DNA damaging agents continue to be the mainstay in various chemotherapeutic regimens used in the clinic. DNA interstrand crosslinks are considered to be the critical cytotoxic lesions for the biological activity of such agents. Gel-based electrophoretic assays can efficiently separate denatured single-stranded DNA from double-stranded, covalently-linked DNA resulting from the presence of an interstrand crosslink. The methods described here offer a simple way for the assessment of crosslinking efficiencies of bifunctional agents in both long fragments of DNA (e.g. 1-5 kb) and short oligonucleotide DNA duplexes. As the repair of interstrand crosslinks is a key determinant of cellular and clinical chemosensitivity, these methods can be useful for the characterization and isolation of site-directed adducted substrates for use in subsequent biochemical analysis of cellular recognition and DNA repair processes. PMID- 19997892 TI - An evaluation cascade for G-quadruplex telomere targeting agents in human cancer cells. AB - The targeting of telomerase and telomere maintenance in human cancer cells can be achieved by small molecules that induce the 3'single-stranded ends of telomeric DNA to fold up into four-stranded quadruplex structures that inhibit the action of the telomerase enzyme complex. In this chapter, we describe a series of biochemical, biophysical, and cellular assays that are used to evaluate the activity of new compounds, and so assess whether they are suitable for examination in xenograft models of human cancer. These assays evaluate quadruplex stabilisation properties, short- and long-term cell viability, telomerase enzymatic activity, cellular senescence, and telomere length changes. PMID- 19997893 TI - [Acute and chronic anal ulcers]. AB - Classic types of anal ulcers are acute and chronic anal fissure. Characteristic symptoms of chronic fissures are severe pain during defecation accompanied by the triad of ulceration, hypertrophic anal papilla and external skin tag. If the symptoms deviate other causes of ulceration must be considered. Primarily, malignancies should be excluded. The special setting in the anal fold, especially with concurrent immunosuppression, could lead to nonspecific manifestations of different proctological, dermatological and infectious diseases, which can only be clarified by further diagnostic workup and histopathology. Only the correct diagnosis will lead to causal and effective therapy. Successfully treated inflammatory dermatoses and precancerous lesions require regular follow-up because a recurrent or persistent course of the disease may result in malignant transformation. PMID- 19997895 TI - [Transmission of multidrug-resistant bacteria in ambulatory settings]. AB - Multidrug-resistant organisms (MDROs) such as methicillin-resistant Staphylococcus aureus (MRSA), vancomycin-resistant enterococci (VRE) und extended spectrum beta-lactamase (ESBL-) producing bacteria are an ever-expanding challenge regarding infection control and prevention strategies also in ambulatory healthcare settings. The most important transmission mode for MDROs is direct or indirect contact involving the hands of healthcare workers. Strict adherence to standard precautions (and especially adherence to alcoholic handrub use) is essential in prevention of cross-transmission. Additional measures contribute to the control of MDROs: These include flagging of patients' records, establishing standards of care for patients with MDROs, continuing education of staff, disclosure of information to other involved healthcare providers and guideline-based antibiotic treatment and prophylaxis. Optimised cooperation of ambulatory healthcare providers and hospitals regarding management and control of MDROs (networking) is a promising future option. PMID- 19997894 TI - [Clostridium difficile-associated infections. How dangerous are the new strains?]. AB - In recent years, a global increase of Clostridium difficile-associated infections (CDI) and the emergence of new hypervirulent strains causing numerous outbreaks was noticed. The appearance of these strains was accompanied by increased morbidity and mortality, affecting patients previously unknown to be at risk for CDI. Meanwhile, these hypervirulent strains occur in Germany as well. So far, the changing epidemiology of CDI did neither change diagnosis nor therapy of CDI but necessitates a series of preventive measures like the surveillance of CDI and contact precautions while caring for affected patients. If there are complicated or severe cases of CDI, the hypervirulent strains should be suspected and stool culture for isolation of C. difficile should be aimed. PMID- 19997896 TI - [Nosocomial infections. Evidence based infection control measures]. AB - Nosocomial infections remain a major side effect of medical care and are at least in part preventable. Evidence-based, valid infection control recommendations are needed based on multidisciplinary development with local adjustment and incorporation into every-day practice. The article reviews the special aspects which have to be considered for producing evidence based guidelines in the field of infection control. However, guidelines designed to prevent nosocomial infections may be ignored even in the face of persuasive scientific evidence. Insufficient compliance to hand hygiene is the most prominent example. Factors which impede the process of translating research findings into infection control practice have to be identified and suggestions how these barriers may be overcome should be provided. PMID- 19997897 TI - [Chronic periaortitis]. AB - Chronic periaortitis is a rare fibroinflammatory disorder which affects the abdominal aorta and may spread into the retroperitoneum, often encasing the ureters. An aneurysma of the abdominal aorta and vasculitis of the thoracic aorta and of supra-aortic vessels may also coexist. Chronic periaortitis can be idiopathic or secondary to different triggers such as drugs, tumors and infections. Abdominal and/or low back pain is the hallmark symptom. Laboratory markers of inflammation are usually increased. The diagnosis rests on computerized tomography or magnetic resonance imaging, which typically show a retroperitoneal mass displacing the aorta anteriorly and the ureters medially. Positron-emission tomography may assist in defining disease activity and extension. Chronic periaortitis should be differentiated from other fibrosing disorders of various origins. Histology is required in atypical cases to secure the diagnosis. Treatment is based on high-dose steroids with a tapering scheme combined with immunosuppressive agents in refractory or relapsing disease. In case of ureter obstruction early DJ-catheter placement is required. Operative interventions to relieve ureter obstruction are rarely necessary provided immunosuppressive treatment is timely instituted. PMID- 19997899 TI - The alternative Pharaoh approach: stingless bees mummify beetle parasites alive. AB - Workers from social insect colonies use different defence strategies to combat invaders. Nevertheless, some parasitic species are able to bypass colony defences. In particular, some beetle nest invaders cannot be killed or removed by workers of social bees, thus creating the need for alternative social defence strategies to ensure colony survival. Here we show, using diagnostic radioentomology, that stingless bee workers (Trigona carbonaria) immediately mummify invading adult small hive beetles (Aethina tumida) alive by coating them with a mixture of resin, wax and mud, thereby preventing severe damage to the colony. In sharp contrast to the responses of honeybee and bumblebee colonies, the rapid live mummification strategy of T. carbonaria effectively prevents beetle advancements and removes their ability to reproduce. The convergent evolution of mummification in stingless bees and encapsulation in honeybees is another striking example of co-evolution between insect societies and their parasites. PMID- 19997898 TI - The ins and outs of mitochondrial iron-loading: the metabolic defect in Friedreich's ataxia. AB - Friedreich's ataxia is a cardio- and neurodegenerative disease due to decreased expression of the mitochondrial protein, frataxin. This defect results in mitochondrial iron-overload, and in this review, we discuss the mechanisms that lead to this iron accumulation. Using a conditional knockout mouse model where frataxin is deleted in the heart, it has been shown that this mutation leads to transferrin receptor-1 upregulation, resulting in increased iron uptake from transferrin. There is also marked downregulation of ferritin that is required for iron storage and decreased expression of the iron exporter, ferroportin 1, leading to decreased cellular iron efflux. The increased mitochondrial iron uptake is facilitated by upregulation of the mitochondrial iron transporter, mitoferrin 2. This stimulation of iron uptake probably attempts to rescue the deficit in mitochondrial iron metabolism that is due to downregulation of mitochondrial iron utilization, namely, heme and iron-sulfur cluster (ISC) synthesis and also iron storage (mitochondrial ferritin). The resultant decrease in heme and ISC synthesis means heme and ISCs are not exiting the mitochondrion for cytosolic use. Hence, increased mitochondrial iron uptake coupled with decreased utilization and release leads to mitochondrial iron-loading. More generally, disturbance of mitochondrial iron utilization in other diseases probably also results in similar compensatory alterations. PMID- 19997900 TI - Case fatality rates after first acute coronary syndrome in persons treated for type 2 diabetes show an improving trend. AB - AIMS/HYPOTHESIS: We analysed whether the prognosis of a first acute coronary syndrome (ACS) in patients treated for type 2 diabetes has improved. We also compared the trends in patients with and without diabetes. METHODS: We used national registers to identify all patients with clinically known type 2 diabetes in Finland during the years 1988 to 2002 (n = 222,940). All first-ever ACS events (n = 43,412) among these patients were identified using the Hospital Discharge Register and the Causes of Death Register. From the National Cardiovascular Disease Register we identified all first ACS attacks (n = 191,403) among non diabetic patients in the country. Finally, we calculated annual age-standardised case fatality rates for ACS for three time periods: prehospital, days 0 to 27 and days 28 to 364 after the first ACS. RESULTS: The case fatality rate of first ACS declined significantly in both sexes at all time points considered. The declining trends were not different between patients with type 2 diabetes and those without. Among men aged 35 to 74 years, 58.5% (95% CI 57.6-59.4%) with type 2 diabetes and 44.1% (95% CI 43.8-44.5%) without diabetes had died from cardiovascular causes 1 year after their first ACS. Among women of the same age, the corresponding figures were 54.2% (95% CI 53.0-55.4%) and 36.5% (95% CI 35.9 37.1%). Men generally had higher case fatality rates than women. However, except for prehospital deaths, diabetic women had the same or even higher case fatality rates than non-diabetic men. CONCLUSIONS/INTERPRETATION: The case fatality rates for first ACS show similar improving trends in patients with type 2 diabetes and in those without. However, case fatality rates have remained higher in patients with type 2 diabetes. PMID- 19997901 TI - Gender and eccentric training in Achilles mid-portion tendinopathy. AB - The role of gender in Achilles tendinopathy is yet to be determined. We hypothesized that female patients respond the same as males to 12 weeks of painful eccentric training. A total number of 75 consecutive mid-portion patients with Achilles tendinopathy (25 females, 38 males) were enrolled in a cohort study with 63 being analyzed after 12 weeks according to their gender for tendon and paratendon microcirculatory mapping. Outcome was determined by pain on visual analogue scale, VISA-A score, Foot Ankle Outcome Score (FAOS), tendon and paratendon capillary blood flow, oxygen saturation, and postcapillary venous filling pressures. Eccentric training resulted in a morning resting pain reduction by 44% in males (P = 0.001) and by 27% in females (P = 0.08). VISA-A score improved in males by 27% from 63 +/- 12 to 86 +/- 13 (P = 0.036) and by 20% in females from 60 +/- 14 to 75 +/- 11 (P = 0.043, P < 0.05 for gender difference). Among females, only one out of five FAOS items was increased (sport 72 +/- 21 to 82 +/- 15, P = 0.045), while in males, four out of five items were increased (symptoms, pain, all-day-life, and sport, all P < 0.01). The microcirculatory gender-specific response to eccentric training revealed a greater postcapillary venous filling pressure reduction among symptomatic females and inconclusive capillary blood flow changes. No change in tendon oxygenation was noted in both genders. Symptomatic females suffering Achilles tendinopathy do not benefit as much as symptomatic males from 12 weeks of eccentric training. The pain reduction is significantly lower among symptomatic females in contrast to males, and the improvement in the FAOS and VISA-A scores is significantly lower among females in contrast to males. Additional treatment options warrant scrutiny to symptomatic females suffering Achilles tendinopathy beyond eccentric training. PMID- 19997902 TI - Use of oral bisphosphonates and risk of venous thromboembolism: a population based case-control study. AB - Oral bisphosphonates are widely used for fracture prevention, but there is a concern regarding potential adverse cardiovascular effects of bisphosphonates. In this large, population-based case-control study, we found no evidence of an association between bisphosphonate use and risk of venous thromboembolism (VTE). INTRODUCTION: We examined the relation between the use of oral bisphosphonates for osteoporosis and the risk of VTE. METHODS: We conducted a population-based case-control study in Northern Denmark (population, 1.7 million). Using the Danish National Registry of Patients, we identified all women with a first-time hospital diagnosis of VTE between 1999 and 2006. For each case, we selected up to ten female population controls, matched on date of the index VTE event and age. Data on use of oral bisphosphonates, other medications, and comorbidity were obtained from medical databases. We used logistic regression to estimate odds ratios (OR) for VTE associated with bisphosphonate users while adjusting for potential confounding factors. RESULTS: Four thousand one hundred ninety-three cases and 41,197 controls were included in the study. One hundred forty-nine cases (3.6%) and 1,078 controls (2.6%) were current bisphosphonate users. The adjusted OR for VTE among the current bisphosphonate users compared with nonusers was 1.03 (95% confidence interval (CI): 0.84-1.26), and when restricted to cases of unprovoked thromboembolism, the adjusted OR was 1.08 (95% CI: 0.82-1.42). There was no association either for pulmonary embolism or for deep venous thrombosis. CONCLUSION: We found no evidence of an association of oral bisphosphonate use with the risk of VTE. PMID- 19997903 TI - Pedometer determined ambulatory activity and bone mass: a population-based longitudinal study in older adults. AB - In this large population-based study, walking was assessed twice yearly for a week, each time by pedometer, had consistent clinically important associations with hip areal bone mineral density (aBMD) in both sexes which appears most important in those over 65 years of age suggesting that walking becomes more important with increasing age. INTRODUCTION: Walking is advocated as a preventative strategy for osteoporosis but the evidence is conflicting in females and lacking in males. The aim of this population-based longitudinal study in community dwelling older people (n=875) was to determine the association between pedometer determined ambulatory activity (PAA) and bone mass. METHODS: Bone mass was assessed as aBMD at the hip and spine using dual X-ray absorptiometry. Steps per day were measured using pedometers for 1 week on four occasions at least 6 months apart. Data were analysed using linear mixed models. RESULTS: At baseline, PAA was positively associated with hip aBMD. An age interaction was present with steps having a stronger association for those aged over 65 years. Longitudinally, the effect of steps on hip aBMD was constant, but not additive over time. For those over 65 years, the difference in hip aBMD between the lowest and highest steps quartiles ranged from 3.1% to 9.4%. With regard to the spine, the relationship between daily steps and spine aBMD was modified by sex. For males; there was no significant relationship between steps and spine aBMD. However, for females, higher steps were associated with higher spine aBMD with the effect being constant over time but not additive. There was no evidence of a threshold effect. CONCLUSION: In conclusion, pedometer-determined ambulatory activity has consistent clinically important associations with hip aBMD in both sexes which appears most important in those over 65 years of age. The associations for spine aBMD were both weaker and inconsistent suggesting site specificity. PMID- 19997904 TI - Epidemiology of hip fracture in Tucuman, Argentina. AB - The incidence of osteoporotic hip fracture was studied previously in a central area of Argentina. Studying Tucuman (north area) was very useful to compare results of the different areas and detect a similar incidence in women and a slightly higher incidence in men compared with previous data for the central region. INTRODUCTION/METHODS: Epidemiology of hip fracture was studied over a 1 year period in the city of San Miguel de Tucuman (SMT) and in the whole province of Tucuman, located in the northeast of Argentina (latitudes 26 degrees and 28 degrees south). The results were compared with previous studies performed in the central region of Argentina. RESULTS: Two hundred and eighty-three patients (208 women and 75 men) aged 50 years or over in SMT suffered a hip fracture. The incidence in females and males was 334.9 and 163.8 hip fractures per 100,000 inhabitants per year, respectively (female/male ratio 2.0). A total of 498 hip fractures were recorded in Tucuman province (367 in women and 131 in men). The results in females and males were 276.5 and 114.7 hip fractures per 100,000 inhabitants per year, respectively. Average age of the female and male population was 78+/-9 and 77+/-9 years, respectively. CONCLUSIONS: These results showed that the incidence of hip fracture in female and male populations in SMT was similar to previous studies performed in the central area of the country. Further studies on the south area of Argentina should be conducted to complete the information on a large country extending from latitudes 22 degrees to 55 degrees S. PMID- 19997906 TI - Perceived treatment, feedback, and placebo effects in double-blind RCTs: an experimental analysis. AB - INTRODUCTION: In double-blind randomized placebo-controlled trials (RCT) of therapeutic interventions, the effects of the treatment may provide feedback that undermines blinding and consequently distorts measures of the effectiveness of the intervention. METHODS AND RESULTS: This possibility was confirmed in an experimental model using a dummy placebo procedure whereby participants were led to believe that they were taking part in testing a cognitive-enhancing drug. In two experiments, false feedback given about cognitive performance influenced participants' beliefs about whether they had been allocated to the active treatment or placebo. These beliefs also appeared to influence actual cognitive performance in that participants who believed they had taken the active treatment had higher accuracy in Experiment 1 and faster reaction times in Experiment 2 than those who believed they had been given a placebo. The addition of no treatment control groups in Experiment 2 showed that these effects could not be accounted for by the feedback manipulation itself, thereby supporting expectancy as a causal factor. DISCUSSION: These results indicate the importance of assessing participants' beliefs about their treatment allocation in real double blind RCTs and considering if and how these may have affected the trial's outcome. PMID- 19997905 TI - A single dose of zoledronic acid reverses the deleterious effects of glucocorticoids on titanium implant osseointegration. AB - This study evaluates the effect of zoledronic acid (ZOL) on the osseointegration of titanium implants in rabbits with glucocorticoid (GC)-induced bone loss, and our findings demonstrated that a single dose of ZOL is able to reverse the detrimental effects of GCs on the osseointegration of titanium implants. INTRODUCTION: The purpose of this study is to evaluate the effect of ZOL on the osseointegration of titanium implants in rabbits with GC-induced bone loss. METHODS: Three groups of six NZW rabbits were treated for 18 weeks with saline (SALINE), GC (methylprednisolone, 0.35 mg/kg three times a week), or GC + ZOL (methylprednisolone + single dose of ZOL, 0.1 mg/kg). The animals received a titanium implant in the left tibia after 6 weeks and were killed at the 18th week. Bone mineral density (BMD) was measured with dual-energy X-ray absorptiometry at baseline, eighth week (W8), and 18th week (W18) after treatment to determine the change upon treatment (BMD). Histomorphometric and serum bone alkaline phosphatase analysis (BAP) were also performed. RESULTS: At W8, GC group had a significant reduction in lumbar spine and tibia BMD compared with SALINE (p = 0.003 and p = 0.000), as also observed for GC + ZOL group (p = 0.014 and p = 0.003) just 2 weeks after ZOL treatment. In contrast, at W18, the GC + ZOL had an evident BMD rescue with similar lumbar spine and tibia BMD compared with SALINE (0.043 +/- 0.006 vs. 0.055 +/- 0.009 g/cm(2), p = 0.457 and 0.027 +/- 0.003 vs. 0.041 +/- 0.011 g/cm(2), p = 0.232) and a significantly higher BMD compared with the GC (p = 0.024 and p = 0.001). Histomorphometry revealed that osseointegration was significantly reduced in GC (tibia cortical thickness and diameter, bone implant contact, total and peri-implant bone area) whereas GC + ZOL had these parameters similar to SALINE (p > 0.05). Likewise, ZOL reversed the BAP alteration induced by GC. CONCLUSIONS: Our findings demonstrated that a single dose of ZOL is able to reverse the detrimental effects of glucocorticoids on the osseointegration of titanium implants, suggesting that ZOL therapy may improve the outcome of bone implants in patients with glucocorticoid-induced osteoporosis. PMID- 19997908 TI - Adverse interaction of warfarin and paracetamol: evidence from a post-mortem study. PMID- 19997907 TI - Forebrain PENK and PDYN gene expression levels in three inbred strains of mice and their relationship to genotype-dependent morphine reward sensitivity. AB - RATIONALE: Vulnerability to drug abuse disorders is determined not only by environmental but also by genetic factors. A body of evidence suggests that endogenous opioid peptide systems may influence rewarding effects of addictive substances, and thus, their individual expression levels may contribute to drug abuse liability. OBJECTIVES: The aim of our study was to assess whether basal genotype-dependent brain expression of opioid propeptides genes can influence sensitivity to morphine reward. METHODS: Experiments were performed on inbred mouse strains C57BL/6J, DBA/2J, and SWR/J, which differ markedly in responses to morphine administration: DBA/2J and SWR/J show low and C57BL/6J high sensitivity to opioid reward. Proenkephalin (PENK) and prodynorphin (PDYN) gene expression was measured by in situ hybridization in brain regions implicated in addiction. The influence of the kappa opioid receptor antagonist nor-binaltorphimine (nor BNI), which attenuates effects of endogenous PDYN-derived peptides, on rewarding actions of morphine was studied using the conditioned place preference (CPP) paradigm. RESULTS: DBA/2J and SWR/J mice showed higher levels of PDYN and lower levels of PENK messenger RNA in the nucleus accumbens than the C57BL/6J strain. Pretreatment with nor-BNI enhanced morphine-induced CPP in the opioid-insensitive DBA/2J and SWR/J strains. CONCLUSIONS: Our results demonstrate that inter-strain differences in PENK and PDYN genes expression in the nucleus accumbens parallel sensitivity of the selected mouse strains to rewarding effects of morphine. They suggest that high expression of PDYN may protect against drug abuse by limiting drug-produced reward, which may be due to dynorphin-mediated modulation of dopamine release in the nucleus accumbens. PMID- 19997909 TI - Effects of diet composition and trophic structure on mercury bioaccumulation in temperate flatfishes. AB - The summer flounder Paralichthys dentatus and winter flounder Pseudopleuronectes americanus support valuable fisheries along the northeastern United States. The importance of these flatfish as a human dietary resource indicates they are potential sources of mercury (Hg) to fish-consuming citizens. In this study, summer flounder (SF) and winter flounder (WF) were collected from the Narragansett Bay (Rhode Island, USA) and were measured for total Hg burden in whole-body or dorsal muscle tissue. Interspecies differences in Hg contamination were analyzed relative to flounder body size, age, and Hg content of preferred prey. Stable isotope signatures were also used to elucidate the effect of trophic processes on Hg accumulation in the estuarine food web. The mean Hg content of SF exceeded concentrations measured in WF across multiple life-history stages (0.039 0.100 and 0.016-0.029 mg Hg/kg wet weight for SF and WF, respectively), and observed values for both species were lower than the US Environmental Protection Agency regulatory threshold of 0.3 mg Hg/kg wet weight. Total Hg concentrations were also positively correlated with flounder age and length, verifying that both flatfish bioaccumulate Hg. SF accumulate Hg at an accelerated rate, however, owing to this species consuming Hg-enriched prey (teleosts, squid, and macrocrustaceans; mean Hg content = 0.023 mg Hg/kg wet weight), whereas WF feed on prey with low Hg levels (amphipods and polychaetes; mean Hg content = 0.013 mg Hg/kg wet weight). The positive correlation observed between mean biota Hg content and stable nitrogen (delta(15)N) isotope signatures further indicates that Hg is trophically transferred through the food web, and higher trophic level organisms (i.e., enriched delta(15)N) have increased Hg concentrations. Therefore, results from this study suggest that dietary preference and trophic structure are the main factors affecting Hg bioaccumulation in the estuary. Total Hg concentrations of flatfish from the Narragansett Bay, however, do not necessarily reflect coastwide contamination patterns. This reinforces the importance of having research conducted at sufficiently small spatial scales, including the local assessment of Hg contamination for the purpose of issuing state consumption advisories. PMID- 19997911 TI - Characterization of thermostable FMN-dependent NADH azoreductase from the moderate thermophile Geobacillus stearothermophilus. AB - The gene encoding an FMN-dependent NADH azoreductase, AzrG, from thermophilic Geobacillus stearothermophilus was cloned and functionally expressed in recombinant Escherichia coli. Purified recombinant AzrG is a homodimer of 23 kDa and bore FMN as a flavin cofactor. The optimal temperature of AzrG was 85 degrees C for the degradation of Methyl Red (MR). AzrG remained active for 1 h at 65 degrees C and for 1 month at 30 degrees C, demonstrating both superior thermostability and long-term stability of the enzyme. AzrG efficiently decolorized MR, Ethyl Red at 30 degrees C. Furthermore, the enzyme exhibited a wide-range of degrading activity towards several tenacious azo dyes, such as Acid Red 88, Orange I, and Congo Red. These results suggested the sustainable utilization of G. stearothermophilus as an azo-degrading strain for AzrG carrying whole-cell wastewater treatments for azo pollutants under high temperature conditions. PMID- 19997912 TI - Symmetrical nail bed uptake on a (99m)Tc-HDP bone scan in a patient with Wegener's granulomatosis. PMID- 19997910 TI - X-ray scattering studies of model lipid membrane interacting with purothionin provide support for a previously proposed mechanism of membrane lysis. AB - Thionins, ubiquitous plant toxins, are believed to act by lysing the membrane of pathogenic organisms. Several competing mechanisms were proposed for the lysis of phospholipid membranes by the toxins. In order to study in more detail the proposed mechanisms and possibly resolve among the competing proposals, the interactions of purothionins with a model lipid membrane in the form of a monolayer were studied. The monolayer formed at the air-water interface was studied by synchrotron X-ray reflectivity and grazing incidents diffraction methods. The model membrane was composed of 90:10 mol% DPPC:DPPS (dipylmitoyl phosphatidylcholine:dipylmitoyl phosphatidylserine). The protein interaction with the monolayer disturbs the in-plane and out-of-plane order of phospholipids, increases the amount of the liquid phase of the monolayer, and increases the average surface area per alkyl chain. The results indicate that the protein is bound only transiently, and after approximately 4 h most of the properties of the monolayer are reminiscent of the pure DPPC monolayer suggesting partial withdrawal of DPPS. Obtained electron density distributions perpendicular to the membrane interface do not show any significant contribution from the adsorbed proteins, further supporting the withdrawal hypothesis. PMID- 19997913 TI - Unsuspected hibernating myocardium detected by routine oncology (18)F-FDG PET/CT. PMID- 19997914 TI - PET/CT for the assessment and quantification of (90)Y biodistribution after selective internal radiotherapy (SIRT) of liver metastases. PMID- 19997915 TI - Vincenz Alexander Bochdalek (1801-1883): some remarks to the article dedicated to his jubilee. PMID- 19997916 TI - Risk factors for malignancy in benign papillomas of the breast on core needle biopsy. AB - BACKGROUND: This study was designed to evaluate the clinical and pathologic parameters of benign papillomas diagnosed on core needle biopsy (CNB) and predict malignancy risk after surgical excision. METHODS: We retrospectively reviewed clinicopathologic findings for 160 CNB-diagnosed benign papillomas followed by surgical excision from 154 patients. RESULTS: Ten (6.3%) of the excised lesions were diagnosed as malignant. Univariate analysis showed that those that were palpable on physical examination, detected as a mass on mammography, or >1 cm on sonography were significantly associated with malignancy. In multivariate analysis, lesions that were palpable (odds ratio (OR), 29.2; 95% confidence interval (CI), 4.06-209.58; P = 0.001) or detected as a mass (OR, 5.68; 95% CI 1.08-29.87; P = 0.04) remained significantly associated with malignancy. In a CART analysis, including all variables, lesions that were palpable and associated with a mass on mammogram were confirmed as malignant. CONCLUSIONS: Breast lesions diagnosed as benign papillomas on CNB had a 6.3% risk of being malignant. The risk was highest for lesions that were palpable and detectable as a mass on a mammogram. In addition, the low-risk patients avoid immediate surgical excision, although they should be followed carefully. PMID- 19997918 TI - Prognostic significance of host- and tumor-related factors in patients with gastric cancer. AB - BACKGROUND: Various factors regarding the biological state of tumors or the nutritional status of patients have been reported individually to correlate with prognosis. Identification of defined patient groups based on a prognostic score may improve the prediction of survival and individualization of therapy. The aim of the present study was to identify clinically useful parameters obtainable before treatment that could be used for predicting clinical outcomes in patients with gastric cancer. METHODS: In 357 consecutive patients who had been treated for potentially curable gastric cancer, we retrospectively analyzed the following clinicopathological factors: sex, age, body mass index, body weight changes, hemoglobin, white blood cell count, neutrophil to lymphocyte (N/L) ratio, serum C reactive protein (CRP), serum albumin, serum cholinesterase, tumor location, tumor size, histology, and clinical tumor node metastasis (TNM) stage. Factors related to prognosis were evaluated by univariate and multivariate analysis. RESULTS: From univariate analysis, significant differences in survival were found for age, hemoglobin, N/L ratio, serum CRP, serum albumin, serum cholinesterase, tumor size, and clinical T and N grouping. N/L ratio, tumor size, and clinical T grouping were identified as independent prognostic indicators in multivariate analysis. A prognostic score was constructed using these variables to estimate the probability of death. The model gave an area under the receiver operating characteristic curve of 0.85 for prediction of death at 5 years. CONCLUSIONS: This model based on N/L ratio, tumor size, and clinical T grouping before treatment offers a very informative scoring system for predicting prognosis of gastric cancer. PMID- 19997919 TI - Breast cancer in young women in a limited-resource environment. AB - BACKGROUND: Despite the higher incidence of breast cancer in young women in developing countries, there is a paucity of data on their management. We present the clinicopathological features and outcome of treatment of women 35 years or younger with breast cancer in a resource-restricted environment. METHODS: A total of 141 patients who were diagnosed with primary breast cancer at 35 years or younger from January 2000 to June 2008 were retrieved from the cancer registry of a breast clinic at a tertiary hospital and a private breast health center in South Africa. Clinicopathological features, treatment, and survival were analyzed. RESULTS: Two patients presented with TNM stage 0 (1.4%), 14 with stage I (9.9%), 47 with stage II (33.35%), 47 with stage III (33.3%), and 31 with stage IV (21.9%). Tumor grade was 3 in 47%, grade 2 in 37%, and grade 1 in 16% of patients. One hundred and four patients with stage 0-III disease underwent treatment with curative intent, 83 had a mastectomy, and 12 had breast-conserving surgery. Ninety patients (86.5%) had chemotherapy, 68 (65.4%) had radiotherapy, and 50 (48.1%) had hormonal therapy. Of 93 patients who completed primary therapy, 4 developed contralateral cancers, 3 had locoregional recurrence, 8 developed synchronous locoregional and distant recurrence, and 19 relapsed with distant metastasis only. The 2-year disease-free and overall survival for stage 0 III disease was 48 and 56%, respectively. CONCLUSIONS: Young women with breast cancer in a resource-limited environment have similar adverse clinicopathological features to those in developed countries. Their disease is more advanced at presentation with poorer outcome. Increased awareness, better systemic therapy, and more comprehensive genetic studies are essential to improve the dismal outcome. PMID- 19997920 TI - Description of sulcal organization of the insular cortex. AB - PURPOSE: To study the morphology of the gyri and sulci of the insular region. MATERIALS AND METHODS: This study examined 20 formalin-fixed cerebral hemispheres (10 right and 10 left). Observations described: the peri-insular sulci, anatomical variations in the insular sulci and gyri, and the relationships between the central insular sulcus and the central cerebral sulcus and the anatomy of the insular arteries. RESULTS: Each insula had a trapezoid shape, surrounded by four peri-insular sulci (anterior, superior, posterior, and inferior). We differentiated the posterior peri-insular sulcus from the inferior peri-insular sulcus. These two sulci have two different axes separated by a clear angle. The central insular sulcus divides the insula into two parts. The anterior insula includes three short gyri and the anterior insular pole. The posterior insula includes two long gyri and the posterior insular pole. This structure defines two intra-insular opercula. In 60% of cases, the superior extremity of the central insular sulcus is in direct continuity with the inferior extremity of the cerebral central sulcus. The superior branch of the middle cerebral artery supplies the majority of the anterior insular gyri, and the inferior branch supplies the majority of the posterior insular gyri. CONCLUSIONS: This study details the morphology of each insular structure (gyri, sulci and poles), data which could facilitate surgical access to this deep cortical area and assist in future work correlating anatomy with function. PMID- 19997921 TI - Symptomatic and quality of life response to tolterodine in subgroups of men with overactive bladder symptoms and presumed non-obstructive benign prostatic hyperplasia. AB - OBJECTIVES: To investigate the symptomatic and quality of life (QoL) response to treatment with tolterodine extended release (ER) in subgroups of male patients with Overactive Bladder Syndrome (OAB) and LUTS suggestive of non-obstructive benign prostatic hyperplasia (BPH) according to age, symptom severity, diabetes mellitus status, and concomitant treatment for LUTS. METHODS: Patients treated with tolterodine ER 4 mg/day for OAB symptoms, alone or added to unsuccessful alpha-blocker treatment of > or =6 weeks duration, and presumed non-obstructive BPH (Q (max) > or = 15 ml/s) were observed for 12 weeks in a non-interventional study. Patients completed the International Prostate Symptom Score (IPSS) and Overactive Bladder Questionnaire (OAB-q) at baseline and after 12 weeks. RESULTS: 52.4% of 741 patients were aged < or =65 years; 4, 64, and 32% had mild, moderate, and severe symptoms, respectively, according to IPSS; 14% had diabetes mellitus, and in 42% tolterodine was added to alpha blockers. In the various subgroups, mean IPSS total scores improved by 2.8-11.1 points, IPSS QoL scores by 1.8-2.4 points, and all OAB-q subscores by more than 14 points. Only IPSS and OAB q baseline scores had a relevant impact on changes during treatment, benefits were greatest in patients with more severe symptoms and bother. CONCLUSIONS: In men with symptoms of OAB and LUTS suggestive of non-obstructive BPH of all IPSS severity classes, aged < or =65 years or above, with or without concomitant diabetes or alpha-blockers, symptoms and QoL improved markedly during treatment with tolterodine ER. PMID- 19997922 TI - [Is bioptic assurance reasonable in patients with Sjogren's syndrome? From focus score to diagnosing vasculitides]. AB - Sjogren's syndrome is an autoimmune disease which targets the salivary and lacrimal glands in particular, causing sicca syndrome. Extraglandular manifestations are often seen. Chronic sialadenitis of the parotid gland is the most common symptom to be assessed for differential diagnosis. Common HE and Giemsa slices are histopathologically examined and graduated for lymphocyte infiltration (focus): grade 0: absent, grade 1: slight, grade 2: moderate non focal infiltration, grade 3: 1 focus (> or =50 lymphocytes) per 4 mm2, grade 4: >1 focus. Grade 3 infiltrates correspond to a focus score of 1, which is one of four disease-classifying criteria acknowledged for diagnosis. Bioptic examination is also performed to rule out different (non-) immunologic sialadenitises, such as the necrotizing or epithelioid-like form (in sarcoidosis), and the extranodal marginal-zone lymphoma. Extraglandular manifestations of Sjogren's syndrome can also be safely diagnosed by histopathological examination. Emphases lie on vasculitides and myositides. Bioptic work-up, therefore, is not only reasonable but also an essential tool for diagnostics in Sjogren's syndrome. PMID- 19997924 TI - Amnestic syndrome due to meningovascular neurosyphilis. PMID- 19997923 TI - Differential nuclear envelope assembly at the end of mitosis in suspension cultured Apium graveolens cells. AB - NMCP1 is a plant protein that has a long coiled-coil domain within the molecule. Newly identified NMCP2 of Daucus carota and Apium graveolens showed similar peripheral localization in the interphase nucleus, and the sequence spanning the coiled-coil domain exhibited significant similarity with the corresponding region of NMCP1. To better understand disassembly and assembly of the nuclear envelope (NE) during mitosis, subcellular distribution of NMCP1 and NMCP2 was examined using A. graveolens cells. AgNMCP1 (NMCP1 in Apium) disassembled at prometaphase, dispersed mainly within the spindle, and accumulated on segregating chromosomes, while AgNMCP2 (NMCP2 in Apium), following disassembly at prometaphase with timing similar to that of AgNMCP1, dispersed throughout the mitotic cytoplasm at metaphase and anaphase. The protein accumulated at the periphery of reforming nuclei at telophase. A probe for the endomembrane indicated that the nuclear membrane (NM) disappears at prometaphase and begins to reappear at early telophase. Growth of the NM continued after mitosis was completed. NMCP2 in the mitotic cytoplasm localized in vesicular structures that could be distinguished from the bulk endomembrane system. These results suggest that NMCP1 and NMCP2 are recruited for NE assembly in different pathways in mitosis and that NMCP2 associates with NM-derived vesicles in the mitotic cytoplasm. PMID- 19997925 TI - Pasteurella multocida meningitis caused by kissing animals: a case report and review of the literature. PMID- 19997926 TI - Concentration of cytokines in age-related macular degeneration after consecutive intravitreal bevacizumab injection. AB - BACKGROUND: To evaluate the changes in aqueous humor cytokine levels following consecutive intravitreal bevacizumab (Avastin, Genentech Inc., San Francisco, CA, USA) injections in eyes with choroidal neovascularization (CNV) due to age related macular degeneration (AMD). METHOD: Aqueous humor samples were collected at the time of intravitreal injection of 1.25 mg of bevacizumab every 7.0 (+/ 2.0) weeks from ten eyes with AMD for the AMD group and during cataract surgery in nine eyes for the control group. Visual acuity with Early Treatment of Diabetic Retinopathy Study (ETDRS) letters and central macular thickness (CMT) using optical coherence tomography were measured before each injection in the AMD group. Aqueous cytokine levels were determined by immunoassay using multi-analyte biochip array technology (Evidence investigator cytokine and growth factor biochip array, RANDOX laboratories Ltd., Crumlin, UK). RESULT: In the AMD group, mean +/- standard deviation(SD) aqueous VEGF levels decreased from 68.0 +/- 32.1 pg/ml at baseline to 26.3 +/- 19.0 pg/ml after the first injection (p = 0.028) and to 25.2 +/- 12.8 pg/ml after the second injection (p = 0.005). While CMT decreased from 307.7 +/- 102.0 mum to 206.8 +/- 141.5 microm (p = 0.037), ETDRS visual acuity increased from 17.6 +/- 11.7 letters to 22.0 +/- 15.6 letters after three consecutive injections (p = 0.017). CONCLUSION: Significantly decreased VEGF levels were noted after the first injection of bevacizumab. These levels were maintained after the second injection, which paralleled the change in visual acuity and CMT. PMID- 19997927 TI - Expression profiling of a complex thaumatin-like protein family in western white pine. AB - The protein content in the plant apoplast is believed to change dramatically as a result of host defense response upon infection with various pathogens. In this study, six novel thaumatin-like proteins (TLPs) were identified in western white pine (Pinus monticola) needle apoplast by a proteomic strategy using two dimensional protein electrophoresis followed by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. Sequent cDNA cloning found that ten P. monticola TLP genes (PmTLP-L1 to -L6 and -S1 to -S4) were expressed in various tissues. Phylogenetic analysis demonstrated that these PmTLP genes belong to a large, complex, and highly diverse plant TLP family. Quantitative reverse transcription-polymerase chain reaction (RT-qPCR) using gene specific primer pairs showed that each PmTLP gene exhibited a characteristic pattern of mRNA expression based on their unique organ distribution, seasonal regulation, and response to abiotic and biotic stresses. A time-course analysis at the early stages of infection by white pine blister rust pathogen Cronartium ribicola revealed that a coordinated upregulation of multiple PmTLP genes was involved in P. monticola major gene (Cr2) resistance. The structural and expressional differentiations suggest that the PmTLP family may contribute to host defense as well as other mechanism. PMID- 19997928 TI - Sex differences in parking are affected by biological and social factors. AB - The stereotype of women's limited parking skills is deeply anchored in modern culture. Although laboratory tests prove men's average superiority in visuospatial tasks and parking requires complex, spatial skills, underlying mechanisms remain unexplored. Here, we investigated performance of beginners (nine women, eight men) and more experienced drivers (21 women, 27 men) at different parking manoeuvres. Furthermore, subjects conducted the mental rotation test and self-assessed their parking skills. We show that men park more accurately and especially faster than women. Performance is related to mental rotation skills and self-assessment in beginners, but only to self-assessment in more experienced drivers. We assume that, due to differential feedback, self assessment incrementally replaces the controlling influence of mental rotation, as parking is trained with increasing experience. Results suggest that sex differences in spatial cognition persist in real-life situations, but that socio psychological factors modulate the biological causes of sex differences. PMID- 19997929 TI - Altered resource availability and the population dynamics of tree species in Amazonian secondary forests. AB - Despite research demonstrating that water and nutrient availability exert strong effects on multiple ecosystem processes in tropical forests, little is known about the effect of these factors on the demography and population dynamics of tropical trees. Over the course of 5 years, we monitored two common Amazonian secondary forest species-Lacistema pubescens and Myrcia sylvatica-in dry-season irrigation, litter-removal and control plots. We then evaluated the effects of altered water and nutrient availability on population demography and dynamics using matrix models and life table response experiments. Our results show that despite prolonged experimental manipulation of water and nutrient availability, there were nearly no consistent and unidirectional treatment effects on the demography of either species. The patterns and significance of observed treatment effects were largely dependent on cross-year variability not related to rainfall patterns, and disappeared once we pooled data across years. Furthermore, most of these transient treatment effects had little effect on population growth rates. Our results suggest that despite major experimental manipulations of water and nutrient availability-factors considered critical to the ecology of tropical pioneer tree species-autogenic light limitation appears to be the primary regulator of tree demography at early/mid successional stages. Indeed, the effects of light availability may completely override those of other factors thought to influence the successional development of Amazonian secondary forests. PMID- 19997930 TI - Endoscopic submucosal dissection for metachronous tumor in the remnant stomach after distal gastrectomy. AB - BACKGROUND: Endoscopic submucosal dissection (ESD) of tumor in the upper part of the stomach is technically difficult and has a high complication rate. We compared the feasibility and safety of ESD for metachronous tumor found in the remnant stomach after distal gastrectomy with ESD for tumor in the upper third of the intact stomach. PATIENTS AND METHODS: We retrospectively compared clinical outcome and safety of 13 lesions (11 early gastric cancers (EGCs) and 2 dysplasias) in the remnant stomach and 30 lesions (28 EGCs and 2 dysplasias) in the upper third of the intact stomach. All patients were treated with ESD at a cancer center hospital. RESULTS: En bloc resection rates were 100% (13/13) for remnant gastric tumor and 87% (26/30) for upper-third tumor (p = 0.30). En bloc resection rates with tumor-free margins (R0 resection) were 92% (12/13) for remnant gastric tumor and 73% (22/30) for upper-third tumor (p = 0.24). Curative resection rates were 85% (11/13) for remnant gastric tumor and 67% (20/30) for upper-third tumor (p = 0.29). Median procedure duration for ESD was 68 min (range 32-233 min) in the remnant stomach and 78 min (range 45-261 min) in the intact stomach (p = 0.49). One case of perforation and one of delayed bleeding occurred in the upper-third tumor group, whereas none occurred in the remnant tumor group. Local recurrence was found in no patients in the remnant gastric tumor group but in one patient in the upper-third tumor group. CONCLUSIONS: ESD for metachronous remnant gastric tumor was feasible and safe in comparison with ESD for tumor in the upper third of the intact stomach. PMID- 19997932 TI - Laparoscopic splenectomy for severe blunt trauma: initial experience of ten consecutive cases with a fast hemostatic technique. AB - BACKGROUND: Minor splenic injuries from blunt trauma can be treated conservatively, whereas high-grade injuries commonly associated with multiple trauma require surgical treatment and usually removal of the organ. Although splenectomy is nowadays routinely performed laparoscopically for the treatment of hematological pathologies, in an emergency the operational procedure is performed through conventional laparotomy worldwide, advocating the need for hemostasis. Progress in surgical skill and new developments in equipment allow us to treat also patients affected by severe splenic blunt trauma minimally invasively. METHODS: In this study we analyzed 12 patients who consecutively came under our observation during a 2-year period and, being affected by severe spleen injury from blunt trauma requiring surgery, underwent emergency laparoscopy. All of them had Injury Severity Score (ISS) >or= 20 with Glasgow Coma Score (GCS) >or= 10. Laparoscopic splenectomy was performed in ten of the cases utilizing a quick hemostatic technique. In one case bleeding was controlled without removal of the organ and in another case laparoscopy revealed that the supposed hemoperitoneum and splenic rupture were in fact the rupture of a giant splenic cyst. RESULTS: The median operative time to reach hemostasis was 17 min (13-125 min) and the median overall operative time was 120 min (55-210 min). All operations were performed fully laparoscopically. Neither mortality nor morbidity related to abdominal problems was observed. Median postoperative stay was 4 days (3-11 days). CONCLUSION: Laparoscopic approach to splenic blunt trauma requiring surgery is a safe and effective procedure. The described technique allows laparoscopic splenectomy to be performed in an emergency, with much the same hemostatic efficacy as the open technique, but with much better outcome for the patient. PMID- 19997933 TI - Surveillance and treatment for second primary cancer in the gastric tube after radical esophagectomy. AB - BACKGROUND: Recent improvement in the survival of patients after esophagectomy for esophageal cancer has led to increasing occurrence of second primary cancer in the pulled-up stomach as gastric tube cancer (GTC). However, a treatment strategy for GTC including surveillance has not been established. The aims of this study are to clarify the incidence and clinicopathological characteristics of GTC and to assess the treatment results of endoscopic resection. METHODS: Twenty-five patients with 29 GTC lesions treated between 1989 and 2007 were analyzed retrospectively. RESULTS: The median interval between esophagectomy and GTC detection was 86 months, and the 10-year cumulative incidence rate of GTC was 8.6%. Of 18 asymptomatic GTCs, 17 lesions (94.4%) were detected by periodic endoscopy and 15 (88.2%) of them were treated endoscopically. Of all 29 GTCs, endoscopic submucosal dissection (ESD) was performed in 10 GTCs with a completely curative resection rate of 90%, which was significantly higher than that of 7 GTCs treated with endoscopic mucosal resection (EMR) (14.3%, P = 0.004). In these 17 GTCs, no cancer recurrence developed during a median follow-up period of 24 months, and the 3-year survival rate was 80.8%. CONCLUSIONS: For patients after esophagectomy with gastric pull-up, long-term follow-up including periodic endoscopy is necessary to detect a potentially curable GTC. ESD is a feasible and safe procedure for GTC, with oncologically favorable features. PMID- 19997934 TI - Dietetic-led management of patients undergoing laparoscopic gastric banding: early results. AB - BACKGROUND: Optimal results of bariatric surgery are achieved when it is performed within a multidisciplinary team. Within this team, the dietician plays a key role before and after surgery in patient education and behaviour change. With long-term follow-up, the number of patients per surgeon increases exponentially. This study evaluated the outcomes of a dietician-only led management program for patients who underwent laparoscopic gastric banding in our unit. METHODS: Between April 2003 and November 2007, 1,335 patients underwent laparoscopic gastric banding in two hospitals by the same surgical team. Weight loss outcomes were compared for patients in a dietician-led management program against a surgeon/nurse specialist follow-up program with more frequent patient visits. For the dietician-led group, a standard protocol of six postoperative visits and two to three fluoroscopic adjustments was developed from referral until 2 years after surgery. RESULTS: There were 316 patients followed up in a dietician-led program. They were compared with the remaining patients who were followed up in a surgeon/ nurse specialist led program. The mean preoperative weight and body mass index (BMI) for the dietetic-led subset was significantly higher (weight: 147.4 +/- 30.2; BMI: 52.8 +/- 8.9) compared with the remaining group (weight: 113.8 +/- 18.7; BMI: 41.6 +/- 5.2; p < 0.001: Mann-Whitney test). Percent BMI loss was initially lower in the dietician-led group, but this difference disappeared at the end of 24 months (p = 0.056). CONCLUSIONS: A patient management program led by specialist dieticians is an effective way to manage large numbers of patients after laparoscopic gastric banding while maintaining comparable weight loss to surgeon/nurse-led series. PMID- 19997935 TI - When does testing for GERD become cost effective in an integrated health network? AB - BACKGROUND: Gastroesophageal reflux (GERD) is the most common gastrointestinal disorder, affecting as many as 14% of the US population. Rising rates of esophageal adenocarcinoma are seen in this population, and chronic proton pump inhibitor (PPI) use does not normalize cancer risk. It has also been demonstrated that up to one-third of patients on PPI therapy did not actually have GERD and could be taken off the medication. These facts form the basis for a quality assurance study of care provided to patients in an integrated health care network who were on high-dose, long-term PPI therapy. METHODS: A cost-benefit analysis of patients who were on double-dose PPI therapy for more than 6 months was performed. Pharmacy, facility, physician reimbursement, and radiologic data from a cohort who were both primary-care patients and insured in our system were utilized. RESULTS: Two hundred and twenty-four patients were prescribed a double dose of this medication for over 6 months. Utilizing a 4.5% discount rate, our break-even analysis showed that Bravo testing [with esophagogastroduodenoscopy (EGD)] needed to identify those patients who could be taken off PPI therapy paid for itself in 33 months. Bravo + EGD + manometry testing needed to screen for other possible pathologies paid for itself in 38 months. Bravo + barium swallow + EGD testing to screen patients for possible esophageal adenocarcinoma paid for itself in 42 months. Bravo + barium swallow + manometry + EGD testing paid for itself in 47 months. CONCLUSIONS: Significant savings can be realized through early use of upper endoscopy, Bravo testing, barium swallow, and manometry to identify patients that are taking double-dose PPIs unnecessarily based on presumptive diagnosis of GERD. This early testing also has the potential to diagnose a variety of other clinically important pathologic conditions more readily. PMID- 19997936 TI - Ischemic conditioning shows a time-dependant influence on the fate of the gastric conduit after minimally invasive esophagectomy. AB - BACKGROUND: Minimally invasive esophagectomy (MIO) is now established as a valid alternative to open surgery for the management of esophagogastric cancers. However, a high incidence of ischemia-related gastric conduit failure (ICF) is observed, which is detrimental to any potential benefits of this approach. METHODS: Since April 2004, MIO has been the procedure of choice for esophagogastric resection in the authors' unit. Data relating to the surgical technique were collected, with a focus on ischemic conditioning by laparoscopic ligation of the left gastric artery (LIC) 2 weeks or 5 days before resection. RESULTS: A total of 97 patients underwent a planned MIO. Four in-patient deaths (4.1%) occurred, none of which were conduit related, and overall, 20 patients experienced ICF (20.6%). In four patients, ICF was recognized and dealt with at the initial surgery. The remaining 16 patients experienced this complication postoperatively, with 9 (9.3%) of them requiring further surgery. Of the 97 patients, 55 did not undergo ischemic conditioning, and conduit failure was observed in 11 (20%). Thirty-five patients had LIC at 2 weeks, and 2 (5.7%) experienced ICF. All seven patients (100%) who had LIC at 5 days experienced ICF. Timing of ischemic conditioning (p < 0.0001) had a definite impact on the conduit failure rate, and the benefit of ischemic conditioning at 2 weeks compared with no conditioning neared significance (p = 0.07). CONCLUSIONS: Ischemic failure of the gastric conduit significantly impairs recovery after MIO. Ischemic conditioning 2 weeks before surgery may reduce this complication and allow the benefits of this approach to be realized. PMID- 19997937 TI - The endoscopic U-stitch technique for primary button placement: an institution's experience. AB - BACKGROUND: Various gastrostomy tube placement techniques have been reported in the literature. The endoscopic U-stitch technique allows for primary button placement without the need for laparoscopy. The purpose of this study was to quantify the completion rate and the occurrence of complications with this procedure at one academic teaching hospital. METHODS: All gastrostomy procedures between February 2001 and September 2008 were reviewed. Data were collected from paper and electronic records for endoscopic U-stitch procedures. RESULTS: Endoscopic U-stitch gastrostomies were attempted in 121 patients, with primary button placement in 115 (95%) and conversion to an open procedure in 6. No procedure-related deaths occurred, and 7% of patients experienced postoperative complications that included hematemesis, cellulitis, colonic perforation, granulation tissue requiring operative excision, and surgical fistula closure. Average operative time was 18 min. CONCLUSION: The endoscopic U-stitch technique is safe and allows for primary button placement in infants and children. Its complication rate compares favorably with other laparoscopic and open techniques described in the literature. PMID- 19997938 TI - Consensus statement of the consortium for laparoendoscopic single-site surgery. PMID- 19997939 TI - Hemolytic uremic syndrome in a preterm infant. PMID- 19997940 TI - Development of renal and iliac aneurysms in a child with generalized infantile myofibromatosis. AB - Infantile myofibromatosis is a rare disorder characterized by the formation of tumors in the skin, soft tissues, bone, and viscera. We report the case of a 3 week-old girl who presented with severe hypertension due to generalized infantile myofibromatosis including renal involvement. The infant was treated by chemotherapy and showed progressive regression of the tumors. However, her evolution was marked by the development of aneurismal dilations of the renal and iliac arteries as observed in fibromuscular dysplasia. We discuss the possibility of a link between these two mesenchymal disorders. PMID- 19997941 TI - Medullary nephrocalcinosis in a pediatric patient. PMID- 19997942 TI - The prokinetic effect of mosapride citrate combined with omeprazole therapy improves clinical symptoms and gastric emptying in PPI-resistant NERD patients with delayed gastric emptying. AB - BACKGROUND: Previous studies have shown that non-erosive reflux disease (NERD) patients are less sensitive to proton pump inhibitor (PPI) treatment than patients with erosive reflux disease. The aim of this study was to investigate whether treatment with prokinetics in addition to omeprazole therapy would improve clinical symptoms, gastric emptying and esophageal peristalsis in PPI resistant NERD patients with or without delayed gastric emptying. METHODS: Subjects were 64 consecutive patients presenting with typical symptoms of PPl resistant NERD (n = 44) and 20 healthy volunteers. PPI-resistant NERD patients underwent mosapride citrate (15 mg/day) and omeprazole (20 mg/day) co-therapy for 12 weeks. We evaluated the clinical symptoms as well as gastric emptying and esophageal manometry before and after combined therapy. We measured both acylated and des-acylated plasma ghrelin levels by the ELISA method. The primary endopoint was to investigate whether co-administration of mosapride citrate and omeprazole would improve clinical symptoms and gastric emptying in PPI-resistant NERD patients with delayed gastric emptying. RESULTS: T (max) value in PPI resistant NERD patients was significantly higher than in healthy volunteers. Combination therapy with the prokinetic agent mosapride citrate and omeprazole significantly improved reflux symptoms and T (max) value in T (max) > 65 min NERD patients. Co-therapy also significantly reduced des-acylated-ghrelin levels in NERD patients with delayed gastric emptying. CONCLUSIONS: Administration of mosapride citrate in addition to omeprazole improved gastro-esophageal reflux and gastric emptying in PPI-resistant NERD patients with delayed gastric emptying. PMID- 19997943 TI - Hepatic encephalopathy: you should only comment on what you have actually measured. PMID- 19997945 TI - New stereoscopic virtual reality system application to cranial nerve microvascular decompression. AB - PURPOSE: Cranial nerve microvascular decompression is a habitual neurosurgical procedure. Authors describe a new application of the Dextroscope (Volume Interactions, Ltd.), a virtual reality environment, to plan and properly simulate this kind of procedures. METHODS: In three cases of hemifacial spasm refractory to drugs and botulinum toxin treatment, the authors used a virtual reality workstation (Dextroscope) to develop an interactive simulation of craniotomy, approach, and Gore-Tex implant optimal size and position in order to reach vascular decompression of facial nerve. RESULTS: Three-dimensional interactive environment allowed the authors to virtually carry out craniotomy, to visualize vascular and nerve relationship, and finally, to select and to simulate best Gore Tex graft positioning in each case. During surgical procedures, facial nerve vascular compressions were exposed and Gore-Tex grafts were successfully placed as it was virtually planned. Patient outcomes were excellent, with an average improvement of two units (0-4 grades of severity scale). CONCLUSION: Virtual reality environment can help the neurosurgeon to plan and train vascular decompression procedures. Use of appropriate craniotomy, knowledge of vascular and nerve relationship, and selection of optimum paddy size and position are the main useful applications of the Dextroscope in these procedures. PMID- 19997944 TI - The traditional Japanese medicine Rikkunshito increases the plasma level of ghrelin in humans and mice. AB - PURPOSE: Rikkunshito is a traditional Japanese medicine that is widely used for treating upper gastrointestinal symptoms. Our purpose is to clarify the effect of Rikkunshito on the levels of peptide hormones and cytokines in healthy humans and mice. METHODS: We administered Rikkunshito, 7.5 g per day, to 21 healthy volunteers for 2 weeks and examined the changes in plasma peptide and hormone levels. In mice, we administered free access to water containing 0, 0.7, 1.1, and 1.4% Rikkunshito for 2 weeks and examined the changes in plasma peptide levels and ghrelin mRNA expression levels in the stomach. RESULTS: Plasma acylated ghrelin levels at 0, 2, and 6 weeks after the start of administration were 6.7 +/ 2.8, 11.7 +/- 4.0, and 10.5 +/- 4.4 fmol/ml, respectively. The levels at 2 and 6 weeks were significantly higher than that at 0 time (p < 0.01, paired t test). The levels of interleukin 4, interferon gamma, and granulocyte colony-stimulating factor in plasma decreased significantly (p < 0.01) after the administration of Rikkunshito. In mice, the levels of plasma acylated ghrelin changed in proportion to the concentration of Rikkunshito in drinking water, and it was revealed by quantitative RT-PCR that the ghrelin mRNA expression level in the stomach was up regulated. CONCLUSION: Rikkunshito increased the plasma acylated ghrelin level in healthy volunteers and normal mice. Additionally, this change was maintained for at least 4 weeks after the end of administration. In mice, Rikkunshito increased the ghrelin mRNA expression level in the stomach. PMID- 19997946 TI - The transclival endoscopic endonasal approach (EEA) for prepontine neuroenteric cysts: report of two cases. AB - INTRODUCTION: The transclival endoscopic endonasal approach was used to completely remove a prepontine neuroenteric cyst in two different patients. CASE REPORTS: Full clinical improvement without postoperative complication was achieved in both cases. The postoperative hospital stay was limited to 2 and 3 days. DISCUSSION: In comparison to posterolateral skull base approaches, the transclival endoscopic endonasal approach allows direct access to the prepontine cistern without unnecessary manipulation of neurovascular structures at the cerebellopontine angle. In contrast to transoral surgery, patients may have decreased risk of infection and can be fed orally immediately without the risks of palatal and oropharyngeal dehiscence. CONCLUSION: Neuronavigation technology, strict adherence to microsurgical principles, and significant endoneurosurgical experience are strongly recommended when approaching these challenging lesions. PMID- 19997947 TI - Structural features of the salt glands of the leaf of Distichlis spicata 'Yensen 4a' (Poaceae). AB - The epidermal salt glands of the leaf of Distichlis spicata 'Yensen 4a' (Poaceae) have a direct contact with one or two water-storing parenchyma cells, which act as collecting cells. A vacuole occupying almost the whole volume of the collecting cell has a direct exit into the extracellular space (apoplast) through the invaginations of the parietal layer of the cytoplasm, which is interrupted in some areas so that the vacuolar-apoplastic continuum is separated only by a single thin membrane, which looks as a valve. On the basis of ultrastructural morphological data (two shapes of the extracellular channels, narrow and extended, are found in basal cells), the hypothesis on the mechanical nature of the salt pump in the basal cell of Distichlis leaf salt gland is proposed. According to the hypothesis, a driving force giving ordered motion to salt solution from the vacuole of the collecting cell through the basal cell of the salt gland to cap cell arises from the impulses of a mechanical compression expansion of plasma membrane, which penetrates the basal cell in the form of extracellular channels. The acts of compression-expansion of these extracellular channels can be realized by numerous microtubules present in the basal cell cytoplasm. PMID- 19997948 TI - Functional consequences of perturbing polyamine metabolism in the malaria parasite, Plasmodium falciparum. AB - Inhibition of polyamine biosynthesis and/or the perturbation of polyamine functionality have been exploited with success against parasitic diseases such as Trypanosoma infections. However, when the classical polyamine biosynthesis inhibitor, alpha-difluoromethylornithine, is used against the human malaria parasite, Plasmodium falciparum, it results in only a cytostatic growth arrest. Polyamine metabolism in this parasite has unique properties not shared by any other organism. These include the bifunctional arrangement of the catalytic decarboxylases and an apparent absence of the typical polyamine interconversion pathways implying different mechanisms for the regulation of polyamine homeostasis that includes the uptake of exogenous polyamines at least in vitro. These properties make polyamine metabolism an enticing drug target in P. falciparum provided that the physiological and functional consequences of polyamine metabolism perturbation are understood. This review highlights our current understanding of the biological consequences of inhibition of the biosynthetic enzymes in the polyamine pathway in P. falciparum as revealed by several global analytical approaches. Ultimately, the evidence suggests that polyamine metabolism in P. falciparum is a validated drug target worth exploiting. PMID- 19997950 TI - Treatment of chronic tophaceous gout with a free flap. AB - A 62-year-old man who suffered from severe deforming arthritis, secondary to chronic tophaceous gout with multilobular, solid, tender, enlarged subcutaneous nodules, and draining tophi in right second finger, was treated by free flap and finally got the more functional and aesthetic result. PMID- 19997949 TI - Influence of surfactants on the effectiveness of bleaching gels. AB - This study evaluated the influence of surfactants on the effectiveness of 35% hydrogen peroxide (HP) and 10% carbamide peroxide (CP) bleaching gels. One hundred and forty bovine teeth were used, which were stained by immersion in a coffee, red wine, and tobacco mixture for 7 days. At the end of this process, the color measurement at baseline was taken with the Vita Easyshade spectrophotometer. The teeth were divided into seven groups: (a) negative control (NC), (b) positive control for HP (PC-35), (c) HP + Tween 20 (T20-35), (d) HP + laurel sodium sulfate (LSS-35), (e) positive control for CP (PC-10), (f) CP + Tween 20 (T20-10), and (g) CP + laurel sodium sulfate (LSS-10). Group NC was kept in artificial saliva for 21 days. Groups PC-35, T20-35, and LSS 35 received three applications of bleaching gel for 10 min; the process was repeated after 7 days. Groups PC-10, T20-10, and LSS-10 received the gel for 8 h per day for 14 days. After the bleaching process, the final color was measured. The analysis of variance and Tukey tests showed statistically significant differences for the parameters of ?L, ?b, and ?E of the HP gels with surfactant and positive control group (PC-35). Within the limits of this in vitro study, the addition of surfactants to HP bleaching gel increased the bleaching effectiveness. PMID- 19997951 TI - Noncontrast multidetector CT of the kidneys: utility of 2D MPR and 3D rendering to elucidate genitourinary pathology. AB - To encourage routine use of 2D multiplanar reconstructions (MRPs) and 3D renderings when interpreting noncontrast computed tomography (CT) of the kidney, as this practice can unveil findings related to the kidney or its vasculature that are not apparent on axial sections. Inspecting the kidneys or characterizing CT findings with both axial sections and MPRs may facilitate identification of vascular and neoplastic pathology. PMID- 19997952 TI - Acute ovarian torsion in an infant: diagnostic clues on supine and decubitus abdominal radiographs confirmed by ultrasound. AB - Acute ovarian torsion is an uncommon yet important diagnostic consideration in any female pediatric patient presenting with acute abdominal pain. A 2-month-old infant presented to the emergency department with a several-day history of constipation. Evaluation with plain radiographs demonstrated a subtle yet persistent soft tissue mass in the right pelvis. Follow-up ultrasound revealed characteristic findings for ovarian torsion and subsequent salpingo-oophorectomy was performed confirming the diagnosis. Acute ovarian torsion is an uncommon and frequently overlooked diagnosis in female infants presenting with gastrointestinal or urinary tract symptoms. Careful assessment of plain radiographic findings may aid in the further management of this difficult diagnosis in female infants. Ultrasound is recommended to confirm the diagnosis. PMID- 19997953 TI - LigaSure versus Milligan-Morgan hemorrhoidectomy: a prospective randomized clinical trial. AB - BACKGROUND: The present study was conducted to compare operative time, postoperative course and outcome of LigaSure hemorrhoidectomy (LH), and conventional open hemorrhoidectomy (OH) for prolapsed hemorrhoids. METHODS: Eighty-four patients with grade III and IV hemorrhoids were randomized into two groups of 42 patients each; group 1 patients underwent LH whereas group 2 patients underwent OH. Data regarding patient demographics, operative details, postoperative pain score, amount of parenteral analgesics required, length of hospital stay, and time until return to work or normal physical activity were all prospectively collected. Postoperative complications and recurrence of prolapse were also recorded. All patients had regular follow-ups every 2 weeks for the first 8 weeks postoperatively, and at 2-month intervals thereafter, for a total period of 12 months. RESULTS: Patient demographics, clinical characteristics, and length of hospital stay were similar in both groups (P > 0.05). The mean operative time, postoperative pain score (up to 48 h), amount of parenteral analgesics required, time off work, and time needed for complete wound healing were significantly less in patients who underwent LH (P < 0.001). Both groups had similar postoperative complications except for delayed wound healing that was observed at 4 weeks postoperatively, in seven patients (16.67%) in the LH group compared to 17 patients (40.48%) in the OH group (chi(2) = 5.83, P = 0.016). Although hemorrhoid recurrence, at 1 year, was also lower among the LH group compared to the OH group (2.38 vs. 9.14%, respectively), the difference was not statistically significant (P = 0.167). CONCLUSIONS: LH is a better alternative than conventional OH in treating prolapsed hemorrhoids (grades III and IV) since it reduces operating time, postoperative pain, and time off work, and allows surgical wounds to heal faster, with minimal comparable side effects and a low recurrence rate. PMID- 19997954 TI - Two port laparoscopic-assisted removal of a migrating rectal foreign body. AB - A variety of rectal foreign bodies have been documented in the English literature. They are inserted for various reasons including social pleasure. Most of them can be removed manually per ano. Here, we report a case of a big soft foreign body in the recto-sigmoid, which required the assistance of laparoscopy to enable retrieval. PMID- 19997956 TI - The macroeconomic impact of pandemic influenza: estimates from models of the United Kingdom, France, Belgium and The Netherlands. AB - The 2003 outbreak of severe acute respiratory syndrome (SARS) showed that infectious disease outbreaks can have notable macroeconomic impacts. The current H1N1 and potential H5N1 flu pandemics could have a much greater impact. Using a multi-sector single country computable general equilibrium model of the United Kingdom, France, Belgium and The Netherlands, together with disease scenarios of varying severity, we examine the potential economic cost of a modern pandemic. Policies of school closure, vaccination and antivirals, together with prophylactic absence from work are evaluated and their cost impacts are estimated. Results suggest GDP losses from the disease of approximately 0.5-2% but school closure and prophylactic absenteeism more than triples these effects. Increasing school closures from 4 weeks at the peak to entire pandemic closure almost doubles the economic cost, but antivirals and vaccinations seem worthwhile. Careful planning is therefore important to ensure expensive policies to mitigate the pandemic are effective in minimising illness and deaths. PMID- 19997955 TI - Headache patients in the emergency department of a Greek tertiary care hospital. AB - The aim of this study was to record the demographic and epidemiological data on adult patients with headache who attend the emergency department (ED) and the diagnoses that made by the neurologists in the ED of a tertiary care hospital in metropolitan Thessaloniki (Greece). In an open prospective study, demographic and epidemiological data were collected on all patients who reported headache (as chief complaint or not) and presented to the ED of Papageorgiou Hospital between August 2007 and July 2008. Headache patients accounted for 1.3% of all ED patients and for 15.5% of patients primarily referred to the ED neurologist. Tension type headache was the most frequent diagnosis, followed by secondary headaches and migraine. The large number of patients without final ED diagnosis and ward admission for further evaluation sheds a light on the immense workload of Greek ED physicians. Furthermore, we found evidence for the misuse of Emergency Medical Services by chronic headache patients. These findings indicate shortcomings in the pre-hospital (primary care) management of headache patients in the Greek National Health System to an extent unreported so far. PMID- 19997957 TI - LIM domain only 4 protein promotes granulocyte colony-stimulating factor-induced signaling in neurons. AB - Granulocyte colony-stimulating factor (GCSF) is currently in clinical trials to treat neurodegenerative diseases and stroke. Here, we tested whether LIM domain only 4 protein (LMO4), a hypoxia-inducible gene that protects neurons from ischemic injury, could modulate the neuroprotective effect of GCSF. We showed that GCSF treatment acetylates and phosphorylates Stat3, activates expression of a Stat3-dependent anti-apoptotic gene, p27, and increases neuron survival from ischemic injury. LMO4 participates in Stat3 signaling in hepatocytes and associates with histone deacetylase 2 (HDAC2) in cancer cells. In the absence of LMO4, GCSF fails to rescue neurons from ischemic insults. In wild-type neurons, inhibition of HDAC promoted Stat3 acetylation and the antiapoptotic effect of GCSF. In LMO4 null cortical neurons, expression of wild-type but not HDAC interaction-deficient LMO4 restored GCSF-induced Stat3 acetylation and p27 expression. Thus, our results indicate that LMO4 enhances GCSF-induced Stat3 signaling in neurons, in part by sequestering HDAC. PMID- 19997958 TI - A thermostable phytase from Bacillus sp. MD2: cloning, expression and high-level production in Escherichia coli. AB - Phytase is used as a feed additive for degradation of antinutritional phytate, and the enzyme is desired to be highly thermostable for it to withstand feed formulation conditions. A Bacillus sp. MD2 showing phytase activity was isolated, and the phytase encoding gene was cloned and expressed in Escherichia coli. The recombinant phytase exhibited high stability at temperatures up to 100 degrees C. A higher enzyme activity was obtained when the gene expression was done in the presence of calcium chloride. Production of the enzyme by batch- and fed-batch cultivation in a bioreactor was studied. In batch cultivation, maintaining dissolved oxygen at 20-30% saturation and depleting inorganic phosphate below 1 mM prior to induction by IPTG resulted in over 10 U/ml phytase activity. For fed batch cultivation, glucose concentration was maintained at 2-3 g/l, and the phytase expression was increased to 327 U/ml. Induction using lactose during fed batch cultivation showed a lag phase of 4 h prior to an increase in the phytase activity to 71 U/ml during the same period as IPTG-induced production. Up to 90% of the total amount of expressed phytase leaked out from the E. coli cells in both IPTG- and lactose-induced fed-batch cultivations. PMID- 19997959 TI - Phase I trial of motexafin gadolinium and doxorubicin in the treatment of advanced malignancies. AB - PURPOSE: To assess the safety, maximum-tolerated dose (MTD), and dose-limiting toxicities (DLT), of motexafin gadolinium (MGd), given in combination with doxorubicin, in patients with advanced solid tumors. STUDY DESIGN: The combination of MGd and doxorubicin was administered every 28 days (cycle 1) and then every 21 days (subsequent cycles). The dose of MGd, given daily for 3 days, was escalated from 1.0 mg/kg/d to 3.3 mg/kg/d, while the dose of doxorubicin was held at 30 mg/m2. RESULTS: Fifteen patients received 37 cycles of treatment, for a median of 2 cycles per patient (range 0-6 cycles). Three patients (20%) completed 6 cycles of therapy. The MTD was identified as MGd, 2 mg/kg/day and doxorubicin, 30 mg/m2. Dose limiting toxicities included grade 3 hypertension, pneumonia, bacteremia, and elevated GGT. Serious adverse events also included pulmonary embolism and urinary tract infection requiring hospitalization. There was no exacerbation of cardiac toxicity. No patients attained a response to treatment. Six patients (54%) had stable disease. The median time to disease progression, or to last assessment, was 49 days (range 8-195 days). CONCLUSIONS: The combination of MGd and doxorubicin was fairly well tolerated. However, due to emerging preclinical data suggesting that MGd inhibits ribonucleotide reductase, further development of the combination of MGd plus doxorubicin is not recommended. PMID- 19997960 TI - A prospective phase II study of cetuximab in combination with XELOX (capecitabine and oxaliplatin) in patients with metastatic and/or recurrent advanced gastric cancer. AB - BACKGROUND: We evaluated the efficacy and safety of cetuximab in combination with XELOX [XELoda(r) (capecitabine) and OXaliplatin] in advanced gastric cancer (AGC) patients. The objectives were to evaluate overall response rate (ORR), progression-free survival (PFS), overall survival (OS), and safety of cetuximab plus XELOX. METHODS: Previously untreated AGC patients received intravenous infusion of cetuximab 400 mg/m2 on day 1 followed by weekly infusions of 250 mg/m2. Oxaliplatin 130 mg/m2 was administered intravenously on day 1 and capecitabine 1,000 mg/m2 bid was administered orally for 14 days of a 3-week cycle. Chemotherapy was given until disease progression or intolerable toxicities. On completing maximum 8 cycles of chemotherapy, patients were allowed weekly cetuximab until progression. Response evaluations were done every two cycles and toxicities were assessed at each visit. RESULTS: Forty-four patients (29 male) were enrolled; median age was 57.5 years (range 36-70). In total, 253 cycles of XELOX chemotherapy (range 1-8, median 6.5 cycles) and 917 cetuximab infusions (range 1-58, median 19.0) were delivered. Overall RR was 52.3%. Median PFS and OS were 6.5 months (95% CI, 4.9-8.4) and 11.8 months (95% CI, 6.7-16.8), respectively. The most common toxicities of all grades were anemia (81.8% of patients), asthenia (81.8%), anorexia (79.6%), hand-foot syndrome (79.6%), acneiform skin eruption (77.2%), and sensory neuropathy (75.0%), and they were mostly grade 1 or 2. Grade 3-4 hematologic toxicities were uncommon (anemia, 6.8%; thrombocytopenia, 2.3%). CONCLUSIONS: Cetuximab in combination with XELOX chemotherapy was active and safe as first-line treatment of metastatic and/or recurrent AGC patients. PMID- 19997961 TI - Temporal evolution of patient characteristics enrolled on phase I trials. AB - PURPOSE: Phase I trials serve a crucial role in anticancer drug development. Given the explosion in the number of both approved anticancer therapies and agents in development, we hypothesized that the characteristics of patients enrolling on phase I clinical trials is evolving. PATIENTS AND METHODS: We reviewed 476 published phase I trials over the past decade encompassing 15,100 patients and determined the following characteristics for patients enrolled: age; percentage with ECOG PS of 0, 1, or 2; sex; race; and number of prior chemotherapeutic therapies received: 0, 1, 2 or >= 3. We also identified the major tumor types enrolled: colorectal, lung, renal, breast, head/neck or "other". The change of patient characteristics over time as well as between the first half of studied period (period 1 = 1998-2001) and the second half period (period 2 = 2002-2006) was analyzed. RESULTS: Colorectal and lung cancer patients together comprise ~35% of all patients enrolled on phase I trials and this has not changed over the past decade. The contribution of "other" malignancies has however significantly increased over time. The proportion of patients with PS2 has declined while that of PS1 has increased. The proportion of patients with >=3 prior therapies prior to study enrollment has also significantly increased. CONCLUSION: The shifting of patient characteristics especially as related to tumor types enrolled and number of prior therapies has important implications for future design of studies and inadequate attention to these issues may slow the accrual process. PMID- 19997962 TI - ATP chemosensitivity testing of new antitumor duplex drugs linking 3'-C ethynylycytidine (ECyd) and 2'-deoxy-5-fluorouridine (5-FdU) in comparison to standard cytostatica and combinations thereof. AB - BACKGROUND: 2'-Deoxy-5-fluorouridylyl-(5'-5')-3'-C-ethynylcytidine [5-FdU(5' 5')ECyd] and 3'-C-ethynylcytidinylyl-(5'->1-O)-2-O-octadecyl-sn-glycerylyl-(3 Omicron->5')-2'-deoxy-5-fluorouridine [ECyd-lipid-5-FdU] are antitumor active duplex drugs and these heterodinucleoside phosphate analogues could be cleaved in vivo by wide-spread phosphodiesterases into different antitumor active antimetabolites. METHODS: We cultured breast MCF-7, MDA-MB-231 and ovarian OVCAR 29 and OAW-42 cancer cell lines and used the luminometric measuring of the ATP tumor chemosensitivity assay to assess the in vitro activity of 5-FdU(5'-5')ECyd and ECyd-lipid-5-FdU in comparison to standard single cytostatic agents and combinations thereof currently used in anticancer therapies. To allow comparison between samples and different regimens IndexSUM was determined based on the percentage tumor cell growth inhibition at each test drug concentration. Additionally, the cytostatic efficacy of 5-FdU(5'-5')ECyd and ECyd-lipid-5-FdU was evaluated at a minimum of five concentrations at 10 fold dilutions using 60 human tumor cell lines including ovarian and breast cancer cell lines from the National Cancer Institute (USA). RESULTS: 5-FdU(5'-5')ECyd and ECyd-lipid-5-FdU have a high cytostatic efficacy reaching 50% tumor cell growth inhibition at concentrations ranging between nano- and micomolar. IndexSum values for broad range efficacy in MCF-7 breast cancer cells were comparable to values obtained for standard drug combinations. Higher cytostatic efficacy was observed in MDA-MB 231 cells. CONCLUSION: The duplex drugs 5-FdU(5'-5')ECyd and ECyd-lipid-5-FdU represent potential new chemotherapeutic drugs for breast and ovarian cancer cells which are comparable to currently used drug combinations and more potent in comparison to some monocytostatica used in cancer therapy. PMID- 19997963 TI - Emerging targeted therapies for bladder cancer: a disease waiting for a drug. AB - Urothelial cell carcinoma is the fifth most common cancer and the costliest to treat. This is largely because of all new cases, about 70% present as superficial disease and this while rarely fatal, tends to recur, requiring long-term follow up and repeat interventions. The standard of care, intravesical chemo- and immunotherapy, while effective, is associated with a considerable side-effect profile and approximately 30% of patients either fail to respond to treatment or suffer recurrent disease within 5 years. Muscle-invasive bladder cancer is life threatening, showing modest chemosensitivity, and usually requires radical cystectomy. Although bladder cancer is fairly well-genetically characterized, clinical trials with molecularly targeted agents have, in comparison to other solid tumors such as lung, breast and prostate, been few in number and largely unsuccessful, with no new agents being registered in the last 20 years. Hence, bladder cancer represents a considerable opportunity and challenge for molecularly targeted therapy. PMID- 19997965 TI - Magnetic fluorescent composite nanoparticles for the fluoroimmunoassays of Newcastle disease virus and avian virus arthritis virus. AB - A new detection format for multiplexed analysis based on the use of magnetic fluorescent composite nanoparticles was presented in this paper. Two different antigens, Newcastle disease virus (NDV) antigen and Avian virus arthritis virus (AVAV) antigen, were conjugated to two kinds of magnetic fluorescent composite nanoparticles of different luminescent colors, while red-emitting CdTe QDs were attached to the antibody of NDV and AVAV. Both CdTe QDs-labeled antibodies and magnetic fluorescent composite nanoparticles labeled antigens were used to form a typical immunoreaction system for the detection of NDV and AVAV. Also a typical mixed separation format was realized, which showed the outstanding magnetic properties of composite nanoparticles and the broad application in immunoseparation. PMID- 19997966 TI - Fluorescence quenching to study protein-ligand binding: common errors. AB - A number of recent articles, amongst others several published in the Journal of Fluorescence, use inappropriate fluorescence methodology to determine ligand binding characteristics to (mostly) proteins. In this Letter, several common pitfalls are discussed in relation to two recent publications in the Journal of Fluorescence (Wang et al. (2009) 19:801-808; Ding et al. (2009) 19:783-791). The Author hopes that this contribution helps to prevent a further spread of the incorrect methodology, and results in a reappraisal of those articles already published using similar methodology. PMID- 19997967 TI - Elevated interleukin-21 correlated to Th17 and Th1 cells in patients with immune thrombocytopenia. AB - BACKGROUND: Interleukin-21 (IL-21) is critical in the development of autoimmune diseases. The role of IL-21 in the pathogenesis of immune thrombocytopenia (ITP) remains unknown. MATERIALS AND METHODS: We examined the expression of IL-21, IL 17, and interferon (IFN)-gamma in ITP patients and controls by enzyme-linked immunosorbent assay and flow cytometry. Detection of specific anti-platelet GPIIb/IIIa and/or GPIb/IX autoantibodies was measured by modified monoclonal antibody specific immobilization of platelet antigens. RESULTS: IL-21 was expressed on both CD3(+)CD8(-) T cells and CD3(+)CD8(+) T cells by flow cytometry. Plasma IL-21 level and the percentage of CD3(+)CD8(-)IL-21(+) T cells and CD3(+)CD8(+)IL-21(+) T cells were significantly elevated in ITP patients compared to controls. The percentage of CD3(+)CD8(-)IL-17(+) T (Th17), CD3(+)CD8( )IFN-gamma(+) T (Th1), and CD3(+)CD8(+)IFN-gamma(+) T (Tc1) cells also significantly increased in ITP patients. Moreover, we found a significant positive correlation between CD3(+)CD8(-)IL-21(+) T cells and Th17 cells. In addition, a positive correlation between CD3(+)CD8(-)IL-21(+) T cells and Th1 cells was also found. CONCLUSION: Together, our results indicated a possible role of IL-21 in ITP patients correlated to Th17 and Th1 cells, and blockade of IL-21 may be a reasonable therapeutic strategy for ITP especially those with active disease. PMID- 19997968 TI - Frequency of Treg cells is reduced in CVID patients with autoimmunity and splenomegaly and is associated with expanded CD21lo B lymphocytes. AB - INTRODUCTION: Common variable immunodeficiency is a heterogeneous antibody deficiency syndrome with autoimmune and inflammatory complications in a significant proportion of patients. The study was designed to evaluate the role of T regulatory (Treg) cells in common variable immunodeficiency (CVID) patients with autoimmunity. METHODS: The number and frequency of Treg cells (CD4(+), CD25(hi), Foxp3(+)) were evaluated in patients and controls, and Foxp3 expression in different subgroups of CVID patients with common clinical manifestations was compared. RESULTS: CVID patients had significantly fewer Treg cells than controls, and low frequency of Treg cells was associated with expansion of CD21(lo) B cells in patients. Patients with autoimmunity had significantly reduced frequency but normal numbers of regulatory T cells, whilst patients with splenomegaly had significant reduction in frequency and number of regulatory T cells. CONCLUSION: Foxp3 is useful on its own or as an adjunct to classify CVID patients although the possibility of reduction in Treg cells as a secondary phenomenon cannot be excluded. PMID- 19997969 TI - Shared agency with parents for educational goals: ethnic differences and implications for college adjustment. AB - This study proposed and confirmed three ways in which college students can perceive shared agency and two ways in which they can perceive non-shared agency with parents when pursuing educational goals in college. Differences and similarities were examined among participants from four ethnic backgrounds (N = 515; 67% female): East Asian American, Southeast Asian American, Filipino/Pacific Islander American, and European American. Results indicated that Asian American youth reported higher levels of non-shared agency with parents (i.e., parental directing and noninvolvement), lower levels of shared agency (i.e., parental accommodation, support, or collaboration), and poorer college adjustment compared to European Americans. However, ethnic similarities were found whereby perceived shared agency in education with parents was associated with college adjustment. Multiple mediation analyses also indicated that our model of shared and non shared agency with parents explained differences in college adjustment between Asian and European Americans, though more strongly for comparisons between European and East Asian Americans. Our results suggest that parents continue to be important in the education of older youth but that continued directing of youth's education in college can be maladaptive. PMID- 19997970 TI - Comparative analysis of American Dengue virus type 1 full-genome sequences. AB - Dengue virus (DENV; Genus Flavivirus, Family Flaviviridae) has been circulating in Brazil since at least the mid-1980s and continues to be responsible for sporadic cases of Dengue fever and Dengue hemorrhagic fever throughout this country. Here, we describe the full genomes of two new Brazilian DENV-serotype 1 (DENV-1) variants and analyze these together with all other available American DENV-1 full-genome sequences. Besides confirming the existence of various country specific DENV-1 founder effects that have produced a high degree of geographical structure in the American DENV-1 population, we also identify that one of the new viruses is one of only three detectable intra-American DENV-1 recombinants. Although such obvious evidence of genetic exchange among epidemiologically unlinked Latin American DENV-1 sequences is relatively rare, we find that at the population-scale there exists substantial evidence of pervasive recombination that most likely occurs between viruses that are so genetically similar that it is not possible to reliably distinguish and characterize individual recombination events. PMID- 19997971 TI - Barking up the wrong evidence tree. Comment on Lurie & Rosenthal, "Concurrent partnerships as a driver of the HIV epidemic in sub-Saharan Africa? The evidence is limited". PMID- 19997972 TI - Seroprevalence of Neospora caninum antibodies and its consequences for reproductive parameters in dairy cows from Dakar-Senegal, West Africa. AB - The aim of this study was to determine the seroprevalence of Neospora caninum antibodies and its effects on reproductive parameters in cows in intensive dairy herds in Dakar. Randomised blood samples were taken for serology from 196 cows in four herds with a history of sporadic abortion. All of the sera were assayed for antibodies against N. caninum, Candida guillermondii, Coxiella burnetii, and Chlamydophila sp. The associations between serostatus and reproductive parameters (abortion, number of inseminations to conception, and calving to conception interval) were assessed over a period of 5 years (2004-2008). The seroprevalence of N. caninum antibodies in dairy cattle was 17.9%. The local Gobra breed and crossbreeds had higher levels of N. caninum antibodies than exotic breeds (p < 0.05). For the most recent pregnancies, seropositive cows required more inseminations to establish conception than seronegative cows (p < 0.05). The results indicate that dairy cattle from Dakar are exposed to N. caninum. Neosporosis should, therefore, be systematically considered as a cause when the calving to conception interval is prolonged. PMID- 19997973 TI - Antioxidant inhibits HMGB1 expression and reduces pancreas injury in rats with severe acute pancreatitis. AB - BACKGROUND: Pathogenesis of severe acute pancreatitis is still unclear, which leads to a lack of proper treatment in severe acute pancreatitis therapeutic strategy. OBJECTIVE: To investigate the effect of treatment with antioxidant pyrrolidine dithiocarbamate on pancreas injury in rats with severe acute pancreatitis and its possible mechanism. METHODS: A total of 144 male Sprague Dawley rats were randomly allocated into a sham operation group (n=48), a severe acute pancreatitis group (n=48), and a pyrrolidine dithiocarbamate-treated group (n=48). All the rats were killed at 1, 3, 6, 12, 24, and 48 h after operation. The pancreas histopathologies were observed and serum amylase levels were tested. Meanwhile, the nuclear factor-kappaB activation, tumor necrosis factor-alpha levels and high-mobility group box protein-1 expression levels in pancreatic tissue were studied. RESULTS: Animals receiving pyrrolidine dithiocarbamate had significantly improved pancreas histopathology and lower serum amylase levels (p<0.05). In the severe acute pancreatitis group, pancreas tumor necrosis factor alpha levels reached a peak at 6 h after operation and afterwards rapidly declined to normal levels. However, high-mobility group box protein-1 levels in pancreatic tissue increased remarkably at the 12th hour, reached a peak at 24 h, and maintained up to 48 h post-severe acute pancreatitis. Compared to the severe acute pancreatitis group, the pancreas nuclear factor-kappaB activity, tumor necrosis factor-alpha, high-mobility group box protein-1 levels in the pyrrolidine dithiocarbamate-treated group all remarkably decreased (p<0.05). CONCLUSIONS: High-mobility group box protein-1 seems to act as a late cytokine mediator in the pathogenesis of severe acute pancreatitis. Pyrrolidine dithiocarbamate might inhibit the activation of nuclear factor-kappaB to blockade tumor necrosis factor-alpha, thereby indirectly suppressing the high-mobility group box protein-1 and reducing pancreatic tissue damage in rats with severe acute pancreatitis. PMID- 19997974 TI - Proinflammatory and anti-inflammatory cytokines present in the acute phase of experimental colitis treated with Saccharomyces boulardii. AB - PURPOSE: To study the proinflammatory and anti-inflammatory cytokines present in the acute phase of trinitrobenzene sulfonic acid (TNBS)-induced experimental colitis treated with Saccharomyces boulardii. METHODS: Thirty male Wistar rats were divided into three groups: (1) treated group--received Saccharomyces boulardii for 14 days; (2) non-treated group--received sodium chloride solution for 14 days; (3) control group. Colitis was induced on the seventh day of the study in the treated and the non-treated groups using TNBS (10 mg) dissolved in 50% ethanol. Quantification of cytokines, including interleukin (IL)-1beta (IL 1beta), IL-6, transforming growth factor-beta (TGF-beta), IL-10 and tumor necrosis factor-alpha (TNF-alpha), in the serum and colonic tissue collected on day 14 were carried out using an enzyme-linked immunosorbent assay (ELISA). RESULTS: The mean concentrations of TGF-beta in both the serum and the colonic tissue of the treated group were statistically higher than that of the control group. The mean concentration of TGF-beta in the colonic tissue of the non treated group was also statistically higher than the control group. CONCLUSION: The group treated with Saccharomyces boulardii showed increased amounts of TGF beta, an anti-inflammatory cytokine, during the acute phase of colitis. There were no differences in the amount of TNF-alpha, IL-1beta, IL-6, and IL-10 between the treated and the non-treated or the control groups during the acute phase of experimental colitis induced by TNBS. PMID- 19997975 TI - Gastric neuromuscular pathology in gastroparesis: analysis of full-thickness antral biopsies. AB - BACKGROUND: Pathologic assessment of gastric tissue in patients with gastroparesis is limited. Aims To evaluate gastric histopathology in patients with gastroparesis. METHODS: Full-thickness antral biopsies were obtained in 28 patients with gastroparesis. Control specimens were obtained from patients undergoing gastric resection. H&E and immunohistochemical stained slides were reviewed for the presence of inflammation, ganglion cells, and interstitial cells of Cajal (ICCs). RESULTS: A mild lymphocytic infiltrate in the myenteric plexus was present in 6 out of 14 patients with diabetic gastroparesis (DG), one of 14 idiopathic gastroparesis (IG) and 0 of eight controls. Significant reductions in total nerve cell bodies were seen in gastroparesis patients (2.2 +/- 0.3 per hpf; p = 0.0002) compared to controls (3.2 +/- 0.12). This was seen in both DG (2.4 +/ 0.32) and IG (2.0 +/- 0.2). Sixteen patients (ten IG and six DG) had a reduction of ganglion cells (<2.3 cells/hpf). C-kit staining showed a reduction of ICCs in six patients (five IG and one DG). Four patients (three IG and one DG) had abnormal ICC morphology on C-kit staining with more rounded morphology and less dendritic processes. CONCLUSIONS: This study shows several pathologic abnormalities in the gastric tissue in some patients with refractory gastroparesis. An inflammatory infiltrate was present in nearly half of the patients with diabetic gastroparesis. There was a reduction in nerve cell bodies in both idiopathic and diabetic gastroparesis. A reduced number of ICCs were found in the myenteric plexus. Thus, histologic abnormalities in gastroparesis are heterogeneous and include myenteric inflammation, decreased innervation, and reduction of ICCs. PMID- 19997976 TI - The redox state of the glutathione/glutathione disulfide couple mediates intracellular arginase activation in HCT-116 colon cancer cells. AB - BACKGROUND: Emerging studies have implicated arginase hyperactivity in the dysregulation of nitric oxide synthesis, which can lead to the development of vascular disease and the promotion of tumor cell growth. Recently, we showed that cysteine, in the presence of molecular iron, promotes arginase activity by driving the Fenton reaction. However, the exact mechanism of arginase activation in the cell induced by oxidative stress is unknown. AIM: The aim of the present study is to examine whether intracellular arginase is regulated by the cellular redox status of glutathione. METHOD: To test this hypothesis, the glutathione/glutathione disulfide redox couple was altered in colon cancer cells with the thiol-specific oxidant, diamide, or the glutathione inhibitor, buthionine-(S,R)-sulfoximine, and the activity of the arginase in the cells was assessed. RESULTS: Treatment of cells with diamide, a thiol-specific oxidant, resulted in a dose-dependent decrease in the glutathione/glutathione disulfide ratio that was associated with the loss of glutathione and a coincident increase in arginase activity and arginase-1 levels in drug-treated cells compared with untreated cells. These results show that oxidation-induced redox changes of glutathione are of sufficient magnitude to control the activity of arginase in the cells. Thus, the physiologic modulation of the glutathione/glutathione disulfide ratio could prove to be a fundamental parameter for the control of arginase activity in pathological conditions of increased oxidative stress. CONCLUSION: This is the first evidence supporting the ex vivo regulation of arginase activity through the redox modulation of intracellular glutathione. The potential adaptive and pathological consequences of glutathione redox regulation of arginase activity are discussed. PMID- 19997977 TI - Effect of atilmotin, a motilin receptor agonist, on esophageal, lower esophageal sphincter, and gastric pressures. AB - BACKGROUND: Motilin, an endogenous gastrointestinal (GI) hormone, increases upper gastrointestinal tract motility and is associated with phase III of the gastric migrating motor complex. The motilin receptor agonist, atilmotin, at doses of 6, 30 or 60 microg intravenously (IV), increases the early phase of gastric emptying. Prior studies at higher doses of 100-450 microg IV demonstrated that some subjects developed noncardiac chest pain. AIMS: The aim of this study is to determine the effects of atilmotin on esophageal, lower esophageal sphincter (LES), and gastric contractility and the development of esophageal-related symptoms. METHODS: Ten healthy volunteers underwent esophageal manometry to study the effects of atilmotin on upper GI motility. Five subjects were studied on three separate days following administration of saline placebo and subsequent IV bolus dose of atilmotin (6, 30 or 150 microg). Another five subjects were studied at the highest dose (150 microg). RESULTS: Atilmotin at 150 microg increased proximal gastric pressure by 6.5 mmHg (P = 0.001 compared with placebo). Atilmotin increased LES pressure at all studied doses; LES pressure increased from 24 +/- 2 mmHg following placebo injection to 34 +/- 4 mmHg following a 30 microg dose of atilmotin (P = 0.007). In the esophagus, atilmotin increased the percentage of failed swallows at the highest dose studied. Failed swallows increased from 17 +/- 7% following placebo injection to 36 +/- 7% following a 150 microg dose of atilmotin (P = 0.016). Atilmotin decreased distal esophageal contractile amplitude only at the highest dose studied, from 69 +/- 8 mmHg (placebo) to 50 +/- 5 mmHg following 150 microg atilmotin (P = 0.018). There were no serious adverse effects or episodes of chest pain with atilmotin. CONCLUSIONS: Atilmotin affects esophageal, LES, and gastric motility. LES and gastric pressures were increased, whereas there was disruption of esophageal peristalsis characterized by lower amplitude and failed contractions. PMID- 19997978 TI - Overexpressing cellular repressor of E1A-stimulated genes protects mesenchymal stem cells against hypoxia- and serum deprivation-induced apoptosis by activation of PI3K/Akt. AB - Bone marrow-derived mesenchymal stem cells (MSCs) have great potential for repair after myocardial infarction. However, poor viability of transplanted MSCs in the ischemic heart has limited their therapeutic potential. Cellular repressor of E1A stimulated genes (CREG) has been identified as a potent inhibitor of apoptosis. The aim of this study was to investigate the anti-apoptotic effects of CREG on MSCs under hypoxic and serum deprivation (SD) conditions. We also investigated the potential mechanism(s) that may mediate the actions of CREG. All experiments were performed on rat bone marrow MSCs. Apoptosis was induced by exposure of cells to hypoxia/SD in a sealed GENbox hypoxic chamber. Effects of CREG were investigated in the absence or presence of inhibitors that target phosphoinositide 3-kinase (PI3K). We found that the overexpression of CREG markedly protected MSCs from hypoxia/SD-induced apoptosis through inhibition of the mitochondrial apoptotic pathway, leading to attenuation of caspase-3. Moreover, CREG enhanced Akt phosphorylation and decreased the expression of p53 in MSCs under hypoxic/SD conditions. The PI3K/Akt inhibitor LY294002 significantly increased the amount of p53 protein and attenuated the anti apoptotic effects of CREG on MSCs. This study indicates that CREG is a novel and potent survival factor for MSCs, therefore, it may be a useful therapeutic adjunct for transplanting MSCs into damaged heart after myocardial infarction. PMID- 19997979 TI - Solitary necrotic nodule of the liver: always benign? AB - OBJECTIVE: Solitary necrotic nodule of the liver (SNNL) is a rare lesion and accepted as a benign entity. The aim of this study is to investigate the possible causes for the development of solitary necrotic nodules. METHODS: Twenty-two retrospective solitary necrotic nodule specimens were examined to evaluate histologic features. The clinical records of these patients were reviewed, and clinical data were obtained for all patients. RESULTS: Histologically, 17 of the 22 nodules were necrotic with surrounding fibrosis, and the remaining five nodules were completely fibrotic. Four of the 22 cases were found to have specific lesions within the nodules which may put light on the pathogenesis. Foci of metastatic carcinoma were identified in two of these four cases, and cuticle fragments of the hydatid cyst were identified in the other two cases. Clinical data showed that half of the cases with solitary necrotic nodule have an associated malignancy mainly involving the gastrointestinal system. CONCLUSIONS: SNNL is not always benign. The possible causes of this lesion include parasites and metastatic tumors. It is important to identify the minute foci of metastatic carcinoma for the appropriate management of this lesion. PMID- 19997980 TI - Non-functional neuroendocrine carcinoma of the pancreas: incidence, tumor biology, and outcomes in 2,158 patients. AB - OBJECTIVE: Pancreatic neuroendocrine cancer is a rare, indolent malignancy with no effective systemic therapy currently available. This population-based analysis evaluated the hypothesis that long-term survival benefit is greater with aggressive, rather than limited, surgical therapy. METHODS: Non-functional pancreatic neuroendocrine carcinoma (NF-pNEC) cases diagnosed from 1973 to 2004 were retrieved from the SEER database. RESULTS: A total of 2,158 patients with NF pNEC were identified, representing 2% of all pancreatic malignancies. The annual incidence increased from 1.4 to 3.0 per million during the study period. On average, tumors measured 59 +/- 35 mm (median 50), and age at diagnosis was 59 +/ 15 years; 29% of patients were younger than 50. Nodal (44%) and systemic metastases (60%) were common. Overall the 5-, 10-, and 20-year survival rates were 33%, 17%, and 10%, respectively. Removal of the primary tumor significantly prolonged survival in the entire cohort (median 1.2 vs. 8.4 years; p < 0.001) and among those with metastases (median 1.0 vs. 4.8 years; p < 0.001). No survival difference was seen between enucleation and resection of the primary tumor (median 10.2 versus 9.2 years, p = 0.456). CONCLUSION: This study suggests that surgical therapy improves survival among patients with localized, as well as metastatic, NF-pNEC. Enucleation may be oncologically equivalent to resection. PMID- 19997981 TI - Acute hyperglycemia worsens hepatic ischemia/reperfusion injury in rats. AB - BACKGROUND/AIMS: Acute hyperglycemia is known to worsen ischemia/reperfusion (I/R) injury following myocardial infarction and stroke. We investigated whether acute hyperglycemia worsens injury and amplifies the inflammatory response evoked by hepatic I/R. METHODS: Rats were pretreated with an intraperitoneal injection of 25% glucose or 0.9% sodium chloride (10 ml/kg BW). Subsequently, rats underwent partial (70%) hepatic ischemia for 45 min. After 4 h of reperfusion, hepatic injury, oxidative stress, inflammation, and heat shock protein expression were assessed. RESULTS: Liver injury was increased in the hyperglycemic group with alanine aminotransferase (ALT) and aspartate aminotransferease (AST) serum concentrations of 7,832 +/- 3,374 and 10,677 +/- 4,110 U/L compared to 3,245 +/- 2,009 and 5,386 +/- 3,393 U/L (p < 0.05 vs. control). Hyperglycemic I/R was associated with increased liver nitrotyrosine concentrations and increased neutrophil infiltration. I/R upregulated the protective heat shock proteins HSP32 and HSP70 in control animals, but this protective mechanism was inhibited by hyperglycemia: HSP32 expression decreased from 1.97 +/- 0.89 (control) to 0.46 +/ 0.13 (hyperglycemia), HSP70 expression decreased from 18.99 +/- 11.55 (control) to 3.22 +/- 0.56 (hyperglycemia), (expression normalized to sham, both p < 0.05 vs. control I/R). CONCLUSIONS: Acute hyperglycemia worsens hepatic I/R injury by amplifying oxidative stress and the inflammatory response to I/R. The increase in injury is associated with a downregulation of the protective heat shock proteins HSP32 and HSP70. PMID- 19997982 TI - Intraperitoneal virtual biopsy by fibered optical coherence tomography (OCT) at natural orifice transluminal endoscopic surgery (NOTES). AB - INTRODUCTION: Fibered optical coherence tomography (OCT) in conjunction with natural orifice transluminal endoscopic surgery (NOTES) could provide a facility for rapid, in situ pathological diagnosis of intraperitoneal tissues in a truly minimally invasive fashion. MATERIALS AND METHODS: A large porcine model was established to test this hypothesis. A standard double channel gastroscope (Olympus) was used to achieve a transgastric access to the peritoneum and initiate the pneumoperitoneum. Magnetic retraction was used to display the sigmoid colon along with its mesentery. A commercially available fibered OCT probe (NIRIS system, Imalux) was inserted via a working channel of the gastroscope and used to assess intraperitoneal tissues. Separately, OCT images of human tissue specimens ex vivo were contrasted with representative standard histopathological slides. RESULTS: Intraperitoneal OCT provided clear real-time images of both the serosal and muscularis propria mural layers as well as the submucosal-muscularis interface. Examination of mesenteric lymph nodes (including sentinel nodes) allowed visualization of their subcapsular sinus. Comparison of representative cross-sections however failed to evince sufficient resolution for confident diagnosis. CONCLUSION: This approach is technically feasible and, if the technology is advanced and proven accurate in human patients, could potentially be used to individualize operative extent prior to definitive resection. PMID- 19997983 TI - An antireflux anastomosis following esophagectomy: a randomized controlled trial. AB - BACKGROUND: Reflux of duodeno-gastric fluid is a significant problem after esophagectomy with gastric conduit reconstruction. Symptoms may be severe and impact considerably upon the quality of life. Previous studies have suggested that a fundoplication type anastomosis may limit post-esophagectomy reflux. AIM: The purpose of this study was to determine whether a modified fundoplication at the gastro-esophageal anastomosis prevents reflux after esophagectomy. METHODS: Prospective multicenter randomized controlled trial to compare a conventional end of esophagus to side of gastric conduit anastomosis with a modified fundoplication anastomosis in patients undergoing esophagectomy with intrathoracic anastomosis. Major outcomes were reflux symptoms, symptoms of dysphagia, and complications. RESULTS: Fifty-six patients were enrolled. The fundoplication anastomosis was associated with significantly lower incidence of reflux (40% vs 70%), as well as a reduced incidence of severe reflux (8% vs 30%). Disturbance of sleep due to reflux was significantly reduced in the fundoplication group (18% vs 47%) as was the incidence of respiratory symptoms. The fundoplication anastomosis was not associated with an increase in dysphagia, and there was no difference in complications between the two groups. CONCLUSIONS: Fundoplication anastomosis during esophagectomy is effective in protecting patients from reflux symptoms after esophagectomy and improves quality of life, particularly with regard to sleep disturbance. PMID- 19997984 TI - Surgery for small bowel perforation in an Asian population: predictors of morbidity and mortality. AB - INTRODUCTION: Peritonitis from small bowel perforation is associated with prohibitive morbidity and mortality rates. The aims of our study were to review our institution's experience in the surgical management of small bowel perforation and to identify factors that could predict morbidity and mortality. METHODS: A retrospective review of all patients who underwent operative intervention for peritonitis from small bowel perforation from January 2003 to May 2008 was performed. Patients were identified from the hospital's diagnostic index and operating records. The severity of abdominal sepsis for all patients was graded using the Mannheim peritonitis index (MPI). All the complications were graded according to the classification proposed by Clavien and group. RESULTS: Forty-seven patients, of median age 68 years (18-95 years), formed the study group. Pneumoperitoneum on chest radiographs was seen in only 11 (23.4%) patients. Foreign body ingestion (17.0%), adhesions (14.9%), and malignancy (12.8%) accounted for majority of the pathologies. There was one patient who had several small bowel perforations from Degos disease. Small bowel resection was performed in the majority of the patients (74.5%). The mortality rate in our series was 19.1%, while another 57.4% patients had perioperative complications. On univariate analysis, American Society of Anesthesiologists score >or= 3, MPI > 26, hypotension, stoma creation, abnormal electrolyte level, and renal impairment were related to worse outcome, while the three independent variables that were related to worse outcome after multivariate analysis were MPI > 26, hypotension, and abnormal serum potassium level. CONCLUSION: Surgery for small bowel perforation is associated with significant morbidity and mortality rates. Patients with more severe peritonitis and physiological derangement were more likely to fare worse. PMID- 19997986 TI - High-density biatrial pacing protects against atrial fibrillation by synchronizing left atrial tissue. AB - PURPOSE: The induction and sustainability of atrial fibrillation (AF) are most commonly due to pulmonary vein (PV) triggers and left atrial (LA) substrate. We hypothesized that simultaneous pacing of the PV ostia, LA, and right atrium (RA) reduces AF susceptibility by synchronizing atrial activity. METHODS: Mongrel dogs were rapidly paced and maintained in AF for 3 months before cardioversion. Pacing leads were then placed at PV ostia, LA, and RA sites. For single-site pacing (SSP), a drive train (S1) and premature stimulus (S2) were delivered from the same electrode. Multisite pacing (MSP) consisted of S1 at all leads simultaneously and S2 at one site. Pacing trials were performed with decremental S2 coupling intervals (CI) to endpoints of AF induction or refractoriness. RESULTS: AF induction was reduced by 40.0% with MSP vs. SSP (7.2% vs. 12.0%, p < 0.001). LA S2 sites were more likely to induce AF than RA sites (11.5% vs. 2.6%, p < 0.0001), with benefit of MSP over SSP at LA sites (8.6% vs. 14.6%, p < 0.0001). The CI for AF induction was longer with SSP vs. MSP (119.4 +/- 13.3 vs. 104.4 +/- 11.2 ms, p < 0.05). Atrial activation times were shortened with MSP vs. SSP (87.4 +/- 4.9 vs. 97.1 +/- 10 ms, p < 0.005). CONCLUSIONS: Atrial synchronization was shown by the decreased activation times with MSP. AF induction was reduced by 40.0% with MSP, supporting the hypothesis of decreased AF susceptibility with synchronization. This is the first study demonstrating AF protection with high-density pacing at LA sites associated with triggers and substrate for the induction and maintenance of AF. PMID- 19997987 TI - Physical fitness in morbidly obese patients: effect of gastric bypass surgery and exercise training. AB - BACKGROUND: There is a growing consensus that bariatric surgery is currently the most efficacious and long-term treatment for clinically severe obesity. However, it remains to be determined whether poor physical fitness, an important characteristic of these patients, improves as well. The purpose of this pilot study is to investigate the effect of gastric bypass surgery on physical fitness and to determine if an exercise program in the first 4 months is beneficial. METHODS: Fifteen morbidly obese patients (BMI 43.0 kg/m(2)) were tested before and 4 months after gastric bypass surgery. Eight of them followed a combined endurance and strength training program. Before and after 4 months the operation, anthropometrical characteristics were measured, and an extensive assessment of physical fitness (strength, aerobic, and functional capacity) was performed. RESULTS: Large-scale weight loss through gastric bypass surgery results in a decrease in dynamic and static muscle strength and no improvement of aerobic capacity. In contrast, an intensive exercise program could prevent the decrease and even induced an increase in strength of most muscle groups. Together with an improvement in aerobic capacity, functional capacity increased significantly. Both groups evolved equally with regard to body composition (decrease in fat mass and fat-free mass). CONCLUSIONS: An exercise training program in the first 4 months after bariatric surgery is effective and should be promoted, considering the fact that physical fitness does not improve by weight loss only. PMID- 19997988 TI - Advantages of multiplex proteomics in clinical immunology: the case of rheumatoid arthritis: novel IgXPLEXTM: planar microarray diagnosis. AB - Clinical multiplex diagnostic proteomics is the application of proteomic technologies to improve a patient's clinical outcomes. The future holds impact potential for testing prognosis, diagnosis, and drug therapy, while monitoring efficacious treatment with qualitative and quantitative data. Multiplex clinical diagnostic use of novel biomarkers in body fluids to confirm presence and severity of clinical disease states, holds great promise for clinical use. Challenges for diagnostic clinics include awareness of proteome complexity in clinical samples, the effects of high-abundance proteins, such as albumin, that could mask detection of other and low abundance disease proteins or biomarkers. Standardized approaches to sample collection and preparation, new analytical techniques and novel algorithms for bio-statistical analysis will facilitate release of the great potential of clinical multiplex diagnostic proteomics. A sensitive RA assay has been developed for the simultaneous measurement of the three rheumatoid factors (RFs), RF-IgA, IgG, and IgM, with the option to simultaneously measure anti-cyclic citrullinated peptide (anti-CCP) IgG antibodies using IgXPLEXTM: technology. Testing 10-MUL serum samples, SQI's multiplex microarray rheumatoid arthritis assay provides both positive/negative as well as qualitative/semi-quantitative results for anti-CCP IgG, RF-IgA, IgG, and IgM in each sample well on a 96-well microtiter-formatted microarray plate. Signal detection uses sensitive fluorescent-tagged markers captured onto planar microarray spots and read in a microarray scanner. Each result is verified with confidence confirmation technology and validating quality controls in every sample well. For an 80-RA positive patient cohort, the 4-PLEX profile sensitivity was determined at 82.5%. The specificity for the 44 RA healthy control cohort was determined at 97.7%. The multiplex data also demonstrated that a patients' severity of disease profile, mild to severe, correlates the status of RA biomarkers to disease status. PMID- 19997989 TI - Atopy risk in infants and children in relation to early exposure to fish, oily fish, or long-chain omega-3 fatty acids: a systematic review. AB - There are two main families of polyunsaturated fatty acids (PUFAs), the n-6 and the n-3 families. It has been suggested that there is a causal relationship between n-6 PUFA intake and allergic disease, and there are biologically plausible mechanisms, involving eicosanoid mediators of the n-6 PUFA arachidonic acid, that could explain this. Fish and fish oils are sources of long-chain n-3 PUFAs and these fatty acids act to oppose the actions of n-6 PUFAs. Thus, it is considered that n-3 PUFAs will protect against atopic sensitization and against the clinical manifestations of atopy. Evidence to examine this has been acquired from epidemiologic studies investigating associations between fish intake in pregnancy, lactation, infancy, and childhood, and atopic outcomes in infants and children and from intervention studies with fish oil supplements in pregnancy, lactation, infancy, and childhood, and atopic outcomes in infants and children. All five epidemiological studies investigating the effect of maternal fish intake during pregnancy on atopic or allergic outcomes in infants/children of those pregnancies concluded protective associations. One study investigating the effects of maternal fish intake during lactation did not observe any significant associations. The evidence from epidemiological studies investigating the effects of fish intake during infancy and childhood on atopic outcomes in those infants or children is inconsistent, although the majority of the studies (nine of 14) showed a protective effect of fish intake during infancy or childhood on atopic outcomes in those infants/children. Fish oil supplementation during pregnancy and lactation or during infancy or childhood results in a higher n-3 PUFA status in the infants or children. Fish oil provision to pregnant women is associated with immunologic changes in cord blood and such changes may persist. Studies performed to date indicate that provision of fish oil during pregnancy may reduce sensitization to common food allergens and reduce prevalence and severity of atopic dermatitis in the first year of life, with a possible persistence until adolescence with a reduction in eczema, hay fever, and asthma. Fish oil provision to infants or children may be associated with immunologic changes in the blood but it is not clear if these are of clinical significance and whether they persist. Fish oil supplementation in infancy may decrease the risk of developing some manifestations of allergic disease, but this benefit may not persist as other factors come into play. It is not clear whether fish oil can be used to treat children with asthma as the two studies conducted to date give divergent results. Further studies of increased long-chain n-3 PUFA provision in during pregnancy, lactation, and infancy are needed to more clearly identify the immunologic and clinical effects in infants and children and to identify protective and therapeutic effects and their persistence. PMID- 19997990 TI - Liver transplantation for primary hepatic leiomyosarcoma: a case report and review of the literatures. AB - Primary hepatic leiomyosarcoma (PHL), a malignant mesenchymal tumor of myogenic origin, is exceedingly rare. Patients may be afflicted with a wide spectrum of nonspecific symptoms, and most of the laboratory data are unhelpful to diagnosis. Here, we report a 44-year-old woman with anorexia and right upper quadrant pain who was diagnosed as having malignant hepatic tumor within the Milan criteria and treated by orthotopic liver transplantation (OLT), the pathologic diagnosis of PHL was made by postoperative immunohistochemical study. She underwent six courses of intravenous chemotherapy and enjoyed a 14-month tumor recurrence-free survival until the lymph node metastasis was detected, then we switched the tacrolimus to sirolimus and the patient survived for 20 months additionally. Unfortunately, she finally died of extensively lymphatic metastasis and cachexia. The diagnostic and therapeutic features of PHL are also discussed by reviewing the literatures. To our knowledge, this is the second report of administering OLT to PHL patients. PMID- 19997991 TI - Study on biodistribution and imaging of radioiodinated arginine-arginine-leucine peptide in nude mice bearing human prostate carcinoma. AB - OBJECTIVE: To investigate the biodistribution and imaging of (131)I-labeled arginine-arginine-leucine (RRL) peptide in human prostate carcinoma bearing nude mice. METHODS: The 10-mer cyclic peptide containing the RRL sequence (YCGGRRLGGC) was synthesized by the solid-phase method. Disulfide bonds between the cysteines maintain the cyclic structure. Radioiodination of the RRL peptide was performed by the chloramine-T method. (131)I-labeled peptides were injected into the nude mice bearing human prostate carcinoma via a tail vein. Biodistribution and imaging results in vivo were obtained. RESULTS: The (131)I-labeling rate of RRL peptide was about 60%. The radiochemical purity was 96.5%. The radiochemical purity of the labeled compound remained 90.3% at 24 h in human blood serum at 37 degrees C. In the biodistribution studies, radiolabeled RRL peptide probe accumulated in the tumor to a level of approximately 2.52 and 0.65% injected dose per gram of tissue at 6 and 24 h after administration. The data for the (131)I labeled control peptide were 0.73 and 0.06% ID/g at 6 and 24 h after administration. The ratios of radioactivity in tumors to radioactivity in blood at 1, 6, and 24 h after injection were about 0.32, 1.12, 1.30 for RRL peptide and 0.30, 0.37, 0.22 for control peptide. The ratios of radioactivity in tumors to radioactivity in muscle at 1, 6, 24 h after injection were about 1.40, 3.94, 9.08 for RRL peptide and 1.98, 2.89, 1.78 for control peptide. At 24 h after administration, the SPECT imaging obtained clearly showed a contrasting tumor on the right armpit of mice with high concentrations of radioactivity, and the surrounding background was very low. CONCLUSIONS: The results suggest that radioiodination of RRL peptide is feasible and that the labeled compound is stable in human blood serum. The (131)I-labeled RRL peptide shows high tumor uptake and good tumor-to-organ ratios that allow noninvasive visualization of tumors. The (131)I-labeled compound is valuable to detect tumors as molecular probe. PMID- 19997992 TI - Altered intracellular Ca2+ regulation in chronic rat heart failure. AB - intracellular Ca(2+) handling by the sarcoplasmic reticulum (SR) plays a crucial role in the pathogenesis of heart failure (HF). Despite extensive effort, the underlying causes of abnormal SR Ca(2+) handling in HF have not been clarified. To determine whether the diastolic SR Ca(2+) leak along with reduced Ca(2+) reuptake is required for decreased contractility, we investigated the cytosolic Ca(2+) transients and SR Ca(2+) content and assessed the expression of ryanodine receptor (RyR2), FK506 binding protein (FKBP12.6), SR-Ca(2+) ATPase (SERCA2a), and L-type Ca(2+) channel (LTCC) using an SD-rat model of chronic HF. We found that the cytosolic Ca(2+) transients were markedly reduced in amplitude in HF myocytes (DeltaF/F(0) = 12.3 +/- 0.8) compared with control myocytes (DeltaF/F(0) = 17.7 +/- 1.2, P < 0.01), changes paralleled by a significant reduction in the SR Ca(2+) content (HF: DeltaF/F(0) = 12.4 +/- 1.1, control: DeltaF/F(0) = 32.4 +/ 1.9, P < 0.01). Moreover, we demonstrated that the expression of FKBP12.6 associated with RyR2, SERCA2a, and LTCC was significantly reduced in rat HF. These results provide evidence for phosphorylation-induced detachment of FKBP12.6 from RyRs and down-regulation of SERCA2a and LTCC in HF. We conclude that diastolic SR Ca(2+) leak (due to dissociation of FKBP12.6 from RyR2) along with reduced SR Ca(2+) uptake (due to down-regulation of SERCA2a) and defective E-C coupling (due to down-regulation of LTCC) could contribute to HF. PMID- 19997993 TI - Developmental iodine deficiency and hypothyroidism impair spatial memory in adolescent rat hippocampus: involvement of CaMKII, calmodulin and calcineurin. AB - Developmental iodine deficiency (ID) leads to inadequate thyroid hormone that impairs learning and memory with an unclear mechanism. Here, we show that hippocampal calcium/calmodulin-dependent protein kinase II (CaMKII), calmodulin and calcineurin are implicated in the impaired spatial memory in adolescent rats following developmental ID and hypothyroidism. Three developmental rat models were created by administrating dam rats with either iodine-deficient diet or propylthiouracil (PTU, 5 or 15 ppm)-added drinking water from gestational day (GD) 6 till postnatal day (PN) 28. Then, the spatial memory to a water maze test was studied in pups before PN42. After testing periods, the latency to platform and the number of error in iodine-deficient and 15 ppm PTU-treatment groups were significantly higher than those in the controls (P < 0.05). Total and phosphorylated CaMKII, calmodulin, and calcineurin in the hippocampus were detected with both the immunohistochemistry and western blotting. Without going through water maze test, iodine-deficient and 15 ppm PTU-treatment groups showed significantly lower CaMKII and calmodulin and significantly higher calcineurin than the controls in hippocampal CA1 and CA3 regions (P < 0.05). After trials of water maze task, however, CaMKII and calmodulin were up-regulated and calcineurin was down-regulated in control group (P < 0.05), but not in iodine-deficient and 15 ppm PTU-treatment groups. Data indicate that hippocampal CaMKII, calmodulin, and calcineurin are involved in the impaired spatial memory induced by developmental ID and hypothyroidism. PMID- 19997994 TI - Gingival fibroblasts established on microstructured model surfaces: their influence on epithelial morphogenesis and other tissue-specific cell functions in a co-cultured epithelium: an in-vitro model. AB - OBJECTIVE: The objective of this study was to investigate how gingival fibroblasts cultured on microstructured model surfaces affect epithelial morphogenesis and other cell functions in a cocultured epithelium while conducting a molecular analysis of interactions between biomaterials employing periodontal tissue cells. MATERIALS AND METHODS: We are the first to have successfully co-cultured gingival fibroblasts together with gingival keratinocytes on biofunctionalized, microstructured model surfaces and, in the resulting co-cultured epithelium, examined the molecules of tissue homeostasis, the differentiation marker keratin (K) 1/10, and involucrin after 1- and 2-week periods of cultivation. Desmoplakin was perceived as evidence of cell-to-cell contact and thus as proof of epithelial integrity. We also analyzed the basement membrane component laminin-5. The aforementioned co-culture model without gingival fibroblasts was used as a control set-up. RESULTS: On the protein level, indirect immunofluorescence demonstrated the presence of K1/10, involucrin and the basement membrane component laminin-5 in the co-cultured epithelium in both culture periods. Furthermore, we observed that these epithelial markers had become re-oriented toward the suprabasal cell layers which, in turn, reflects the native in-vivo gingival epithelium. We identified cell-to-cell adhesion as a function of desmoplakin after just 1 week. In the mRNA analysis using quantitative RT-PCR after 2 weeks of cultivation, we noted a considerable rise in relative gene expression that was time-dependent for the early keratinocyte differentiation marker K1 and late marker involucrin. CONCLUSIONS: Our findings demonstrate that gingival fibroblasts on microstructured model surfaces are vitally important for tissue- specific cell functions such as epithelial morphogenesis and other biological cell functions such as differentiation and epithelial integrity. These study findings can thus contribute to the optimization and/or new development of biomaterials currently used in dental medicine. PMID- 19997995 TI - Abrasive effect of air-powder polishing on smoothsurface sealants. AB - AIM: To prevent decalcification during orthodontic treatment using fixed appliances, many orthodontists use sealants to protect the enamel surface around the brackets. If the patients' oral hygiene is insufficient, air polishing devices are often used to clean the teeth. The aim of this study was to investigate the effects of air polishing on the sealant. MATERIAL AND METHODS: Three sealants (Pro Seal, OrthoSolo, and Seal&Protect((R)) were used in this study. Each was applied on the surface of 30 extracted human teeth. The enamel surfaces of 10 teeth each were then air polished with the sodium-bicarbonate based powder (Air-Flow((R))) 5 or 10 seconds, or the glycine-based powder (Clinpro Prophy) for 10 seconds. RESULTS: The sealant had become thinner, revealing minor defects after 5 seconds of air polishing using the sodium bicarbonate-based powder, practically disappearing after 10 seconds. Use of the glycine-based powder allowed longer air-polishing times, but the sealant surface also showed minor defects thereafter. CONCLUSION: Reliable protection against demineralization may be seriously compromised after sealed tooth surfaces are cleaned with air-polishing devices. This finding correlates with the type of powder and air-polishing time. PMID- 19997996 TI - In-vitro study of surface changes in fixed orthodontic appliances following air polishing with Clinpro Prophy and Air-Flow. AB - OBJECTIVE: Air-polishing devices (APDs) are used to clean and polish tooth surfaces. The objective of this study was to examine the effects of exposing orthodontic wires and brackets to different types of powder when using an APD. MATERIAL AND METHODS: Three different wire materials (steel, TMA, Ni-Ti) were polished using an APD (Air-Flow((R)) S1), Clinpro Prophy and Air-Flow((R)). The exposure times were set at 5 and 10 seconds, the application distance at 2 and 4 mm. The wires were then measured for friction. The unpolished sections of the wires were used for control purposes. In a qualitative analysis, the brackets (steel, ceramic and plastic) were exposed for 5 s, 10 s, 30 s und 60 s from a distance of 2 mm using Clinpro Prophy and Air-Flow((R)). The polishing effects were then observed using a scanning electron microscope. RESULTS: The unpolished TMA wire exhibited greater friction (1.22 N) than the Ni-Ti (0.82 N) and the steel wire (0.97 N). Polishing resulted in a 2.5% increase in friction on average (p = 0.004). There was no evidence of a change in the ceramic bracket surfaces. When the Air-Flow((R)) device was used on steel brackets for longer exposure times, the result was a slight roughening of the surface. The plastic brackets exhibited changes that became more obvious after use of the Air-Flow((R)) device. CONCLUSIONS: The surface changes in archwires and brackets caused by air polishing devices when applying the Clinpro Prophy and Air-Flow((R)) powders can be considered as marginal overall. When the adjacent periodontium is intact, both powders are equally suitable, although the Air-Flow((R)) powder is somewhat more effective due to its greater abrasiveness. When using plastic brackets, Clinpro Prophy proved to be the powder of choice, thanks to its low abrasiveness. PMID- 19997997 TI - Torque stability of plastic brackets following multiple loading and artificial material aging--an in-vitro comparison. AB - OBJECTIVE: The objective of this study was to compare the loaddeflection behavior of plastic brackets made of various materials in response to repeated torque loads with each other and with steel brackets. Material fatigue during wire's play in the bracket slot, the brackets' elasticity and the torsional forces applied were analyzed. MATERIAL AND METHODS: Groups of ten brackets, each made of pure polycarbonate or variously reinforced polycarbonate or polyurethane, with and without a metal slot, were artificially aged and then torqued with a testing machine five times consecutively in a torque- measuring apparatus. The control group consisted of ten steel brackets. The resulting forces were recorded with the testing machine, and the wire's deflection recorded with a digital goniometer on a PC and submitted to one-way variance analysis at p < 0.05. RESULTS: There were strong fluctuations among the bracket types in both the play of the wire in the bracket slot as well as the brackets' elasticity. The slot of all the polycarbonate-based brackets was bent open after a single load, except for those with a metal slot. After a single load of up to 20 degrees torque, all the brackets exhibited a significant loss of torque stability ranging between 5% for pure polyurethane and 28.5% for ceramic-reinforced polycarbonate. The loss of torque stability was roughly 17% on average. This loss did not increase significantly when additional loads were applied. CONCLUSIONS: Each bracket material requires its own torque value to transfer identical torque values onto the tooth in clinical practice. Comparison with steel brackets revealed that only plastic brackets with a metal slot are suitable for clinical use. Adding ceramic and glass fibers to polycarbonate, or using polyurethane has no benefit in terms of torque stability. In addition, after a single application of torque, all brackets lose torque stability in response to a renewed load. PMID- 19997998 TI - Studies on the craniofacial morphology of adult cleft patients using euclidean distance matrix analysis (EDMA): a cephalometric study. AB - OBJECTIVE: The aim of this study was to compare the craniofacial morphology of a group of orthognathic adults to adult patients with unilateral cleft lip and palate who had undergone delayed cleft closure. PATIENTS AND METHODS: Lateral cephalograms of 38 adult patients with unilateral cleft lip and palate (cleft group) were submitted for cephalometric analysis with Onyx Ceph((R)) and the x-y coordinates compared to measurements from a reference group of 103 untreated orthognathic adults (non-cleft group). Comparison was based on the cephalometric variables, and a Euclidean distance matrix analysis (EDMA) was applied to identify morphological differences, and describe their size and location. RESULTS: Cephalometric analysis revealed a mesial basal jaw relationship with a retrognathic craniofacial morphology in the cohort with unilateral cleft. EDMA revealed an increase in the vertical distances between A-Pog, ANS-Pog and N-Or. Deficits in the sagittal and vertical distances of the midface (S-ANS, Ba-A) were more pronounced. Differences in the mandibular morphology were also apparent (Ba Go). CONCLUSIONS: Key morphological characteristics of cleft patients are pronounced underdevelopment of the midface in vertical and sagittal dimensions, as well as shortening and clockwise rotation of the mandible. Therefore, bimaxillary procedures should be con- sidered for the purposes of surgical correction, rather than correction of the maxillary position alone. PMID- 19997999 TI - Assessment of quality of life in patients undergoing orthognathic surgery. AB - CLINICAL QUESTION: Analysis to date on the impact of a pronounced skeletal dentofacial deformity on quality of life remains inadequate. OBJECTIVE: The objective of this study was to introduce a validated and internationally comparable questionnaire for the assessment of quality of life, and to use this questionnaire in a consecutively-selected patient cohort. MATERIAL AND METHODS: The Orthognathic Quality of Life Questionnaire (OQLQ) used internationally was translated into German and adapted in accordance with EU guidelines (OQLQ-G). A total of 50 patients (f = 25, m = 25) indicated for combined orthodontic and surgical treatment were included in the study. In addition to the assessment of individual questions, the patients were classified according to the following categories: (1) perception of the malocclusion, (2) masticatory function, (3) esthetics, and (4) social aspects. The statistics were assessed with software SPSS((R)) 15.0 (chi-squared test; p = 0.05). RESULTS: Patients with a pronounced dentofacial malocclusion demonstrated a considerably high awareness of the degree of the deformity. Responses frequently included severe and very severe functional restrictions (50.4%) and/or esthetic impairment (43.0%). With regard to social aspects, the responders also reported a reduction in quality of life. CONCLUSION: An internationally-comparable assessment of quality of life in patients with a pronounced jaw discrepancy was carried out for German-speaking countries. This pilot study demonstrates that their quality of life is significantly impaired. PMID- 19998001 TI - [Quality demands on the assessment of colour vision]. AB - Assessing colour vision comprises a wide spectrum of methods, many of which are practical and highly informative. Given this methodological variety this review aims to help select the most appropriate test and how to correctly execute it, thus achieving the highest quality.Some aspects of the physiology of colour vision are covered as far as is necessary for a basic understanding of colour testing methodology and possible pitfalls. For congenital colour anomalies most pertinent are questions of occupational aptitude. For acquired colour deficiencies assessing colour vision supplements diagnostics of the retina and the visual pathway, allowing both early diagnosis and/or monitoring. For both these fields colour tests provide different kinds of evidence and need to be adequately selected. Methodical artefacts due to both equipment design properties and testing procedures are highlighted so they can be avoided. A form is presented for recording colour examination results commensurate with quality objectives. Finally, a tabular overview of 19 common colour vision tests is provided. PMID- 19998004 TI - [On the press coverage on the Austrian allocation of vaccine against the pandemic influenza A/H1N1]. PMID- 19998005 TI - ["Spare time for research" -- a reality in Austria's University clinics]. PMID- 19998006 TI - Oral anticoagulation with coumarins for patients with atrial fibrillation and chronic kidney disease? PMID- 19998007 TI - [Pandora's Box: pathogens in Ixodes ricinus ticks in Central Europe]. AB - Among the various species of hard ticks, Ixodes ricinus is the most frequently found tick throughout Europe. As with other ixodid ticks, the developmental cycle runs through three stages. In each stage a blood meal is required in order to develop to the next stage. Ixodes ricinus has been found to feed on more than 300 different vertebrate species. Usually, larval ticks feed on small mammals such as mice and become infected with various microorganisms and viruses, of which some are substantial pathogens to humans. The pathogens remain in the tick during molting and are thus transstadially transmitted to the next developmental stage. Pathogens transmitted to humans are the agents of Lyme borreliosis, the tick borne encephalitis virus, Rickettsia species, Anaplasma phagocytophilum, occasionally Francisella tularensis, and protozoal Babesia species. Within the scope of an EU project Ixodes ricinus ticks from all federal states of Austria were searched by means of PCR methods for bacterial pathogens such as Anaplasma phagocytophilum, Borrelia burgdorferi sensu lato, Coxiella burnetii, Ehrlichia spp., Francisella tularensis, Rickettsia spp., and protozoal Babesia. Additionally, the prevalence of Bartonella spp. in this tick species was also determined. Besides the singular detection of Coxiella burnetii and Francisella tularensis in one tick collection site the overall prevalence of Anaplasma phagocytophilum, borreliae, rickettsae and babesiae in Ixodes ricinus amounted to 15%, 14%, 6% and surprising 36% and 51%, respectively. Bartonellae were detected in about 7%. PMID- 19998008 TI - An unusual pattern of Ecchymosis related to Gua Sha. PMID- 19998009 TI - Scientific activity and working hours of physicians in university hospitals: results from the Innsbruck and Salzburg physician lifestyle assessment (TISPLA). AB - OBJECTIVE: Controllable lifestyle has become an important factor influencing career decision-making among physicians. In academic medicine, doctors are required to combine both patient care and research in their daily routine. Insufficient release of clinicians for research during contracted work hours may lead to increased weekly working hours in academic medical centers and deter medical graduates from academia. We tested for an association between numbers of scientific publications and an increased hourly workload among physicians. METHODS: This was a cross-sectional online survey among all salaried physicians working in the university hospitals of Innsbruck and Salzburg, Austria. The main outcome measures were the self-reported total number of scientific papers published in peer-reviewed medical journals over the past two years and self reported working hours. RESULTS: Of 590 returned surveys, 393 were fully completed and included in the study. The sample was stratified into three groups according to scientific output in the past two years: Group A, >/= 6 publications; Group B, 1-5 publications; Group C, no publications. Men were more likely than women to have a scientific publication: in Group A there was a male predominance of 75%, whereas in Group C only 48% were men (P = 0.0034). A total of 59% (n = 232) of all participants had not published a scientific article in the past two years (Group C) and worked a mean of 58.3 +/- 12 h/week. Physicians in Group B (n = 113) had published 2.4 +/- 1.4 papers and worked 62.8 +/- 12.9 h/week; those in Group A (n = 48) had published 11.5 +/- 6.6 papers and worked 73 +/- 13.1 h/week (P < 0.0001). In Group A, research accounted for only 13.3% of total work time but for 60% of overtime hours, reflecting the fact that research was mainly performed during overtime. CONCLUSION: Research activity among clinicians in academic medical centers is associated with significantly increased overtime hours. Measures need to be taken to allow medical graduates an academic career at reasonable impairment of personal lifestyle. PMID- 19998010 TI - [Rationale for bone marrow examination in patients with inflammatory rheumatic diseases]. AB - OBJECTIVE: Inflammatory rheumatic diseases and the applied immunosuppressive treatments can lead to bone marrow depressions and promote hematologic malignancies. Our objective was to explore indications for and results of bone marrow examinations in a large cohort. METHODS: Between 1990 and 2004 146 bone marrow examinations in 3638 patients were performed due to abnormal laboratory results. Medical history, results of bone marrow examination (morphology, histology) and cytogenetic data were investigated retrospectively. RESULTS: Patients' (67.8% female) mean age at bone marrow examination was 53.5 years (SD 15.5), median disease duration 2.9 years. Indications for bone marrow examination were changes in peripheral blood counts in 81.7%. In 52 patients (35.6%) clinically relevant, partially neoplastic bone marrow changes (5 non-Hodgkin lymphoma, 9 myelodysplastic syndromes (MDS)/acute myeloid leukemia (AML) and 3 myeloproliferative neoplasias) were evident. Medication history showed intake of hydroxy-/chloroquine (13.5%), methotrexate (17.3%), cyclosporin (7.7%), sulfasalazine (7.7%), mycophenolatmofetil, gold, leflunomide (each 1.9%), azathioprine (aza, 25.0%) or cyclophosphamide (cyc, 7.7%) prior to bone marrow examination. 7 out of 9 patients, who developed MDS/AML had been treated with either azathioprine alone or additionally with cyclophosphamide (n = 3). CONCLUSION: One third of our patients showed relevant bone marrow changes that might be associated to therapy. The risk seems to be increased especially in patients with inflammatory rheumatic diseases who had received azathioprine alone or in combination with cyclophosphamide. Health care providers should bear in mind the risk of hematologic malignancies and monitor patients closely in this respect. Bone marrow examination should be performed in case of changes in peripheral blood counts; especially clinically relevant anemia, granulocytes < 2,500/microl, thrombocytes < 100,000/microl and relevant changes over time should lead to bone marrow examinations. PMID- 19998011 TI - Atrial fibrillation is an independent determinant of increased NT-proBNP levels in outpatients with signs and symptoms of heart failure. AB - BACKGROUND: N-terminal pro-B-type natriuretic peptide (NT-proBNP) is increasingly used in the diagnosis and prognostic assessment of heart failure; however, the possible influence of atrial fibrillation on BNP is still a matter of controversy. We assessed the influence of atrial fibrillation on NT-proBNP levels in outpatients with signs and symptoms of heart failure. METHODS: Consecutive outpatients (n = 306) referred to a university hospital heart-failure clinic for evaluation of signs and symptoms of heart failure underwent clinical and echocardiographic assessment and had their NT-proBNP levels determined in a sandwich chemiluminescent immunoassay with two antibodies on an Elecsys analyzer. The influence of atrial fibrillation on NT-proBNP levels was assessed using a non parsimonious linear regression model with propensity score adjustments to balance for possible confounders. RESULTS: Atrial fibrillation was associated with increased NT-proBNP levels in patients with (median concentration 1944 vs. 1390 pg/ml) and without (1093 vs. 172 pg/ml) underlying structural disease (P < 0.001). In a linear regression model with a propensity score, atrial fibrillation emerged as an independent determinant of NT-proBNP levels (P = 0.023), even after allowing for possible confounders (left ventricular ejection fraction and end diastolic diameter, left atrial diameter, mitral insufficiency, age, sex, NYHA class or heart rate). CONCLUSIONS: Atrial fibrillation is an independent determinant of increased NT-proBNP levels. This association should be taken into account when NT-proBNP levels are used in the diagnosis of heart failure in patients with atrial fibrillation. PMID- 19998012 TI - Robot-assisted laparoscopic cardiomyotomy. AB - INTRODUCTION: Robot-assisted laparoscopic surgery is an expanding field of medicine. In endoscopic microsurgery particularly, difficult maneuvers such as intracorporeal hand-sewn anastomoses or a narrow operating field are ideal indications for use of an operation robot. PATIENTS AND METHODS: Six patients, four men and two women, underwent robot-assisted laparoscopic cardiomyotomy for achalasia at a tertiary referral center with substantial expertise in robotic surgery. A detailed description of the operation technique is provided, together with a review of the literature. RESULTS: Robot-assisted laparoscopic cardiomyotomy was feasible without any particular problem and the postoperative course of all six patients was uneventful. The operation time was 236 (220-316) minutes, plus 38 (25-47) minutes for setup-time of the robot. At follow-up six months postoperatively, five of the six patients were free of significant dysphagia and all were free of reflux symptoms. DISCUSSION: There are several published reports, series and trials on robot-assisted laparoscopic cardiomyotomy. The general conclusion is that in experienced hands this operation is easy to perform, with a significantly lower rate of mucosal perforations, but that overall costs are higher, including a longer operation time during the learning curve. The avoidance of mucosal lacerations and their possible consequences has to be weighed against higher overall costs. CONCLUSION: Laparoscopic cardiomyotomy is the first standard laparoscopic operation where a clear advantage for use of an operation robot has been proven. Thus, wherever an operation robot is available it should be used for this procedure. PMID- 19998013 TI - Treatment of inappropriate sinus tachycardia with ivabradine. AB - BACKGROUND: Inappropriate sinus tachycardia is most often treated with beta blockers; in resistant cases, nonpharmacologic treatment has been attempted. Recent case reports have shown a favorable response to ivabradine in some patients. METHODS: A total of 13 patients (11 women, 2 men) aged from 27 to 66 years (mean 42 +/- 8) and having inappropriate sinus tachycardia were treated with ivabradine 15 mg per day. RESULTS: In 12 patients whose previous therapy could be discontinued or who did not have previous medication the mean daily heart rate decreased from 94.0 +/- 10.0 to 74.6 +/- 5.2 bpm (mean +/- SD) after ivabradine treatment. In 10 patients in whom we could reliably measure the highest and lowest daily heart rates, the highest rate decreased from 150.3 +/- 13.4 to 120.6 +/- 9.8 and the lowest from 66.7 +/- 9.6 to 54.8 +/- 6.9. All of the differences were statistically significant (P < 0.001). The percentage of heart rate reduction correlated with the initial heart rate (P < 0.005). The remaining patient, who had been treated with metoprolol 300 mg per day for more than 10 years and still had a resting heart rate of 106 bpm, switched to 15 mg of ivabradine daily without discontinuation of drug therapy. After this switch, a decrease of 4 bpm was noted in her mean daily heart rate. CONCLUSIONS: The results of our study show that patients with inappropriate sinus tachycardia can be successfully treated with ivabradine. PMID- 19998014 TI - Recurrent Pott's puffy tumor - atypical presentation of a rare disorder. AB - We report the case of a 14-year-old girl who presented with an 8-week history of unilateral pansinusitis. In the course of the disease a right-side frontal swelling was observed, which disappeared following antibiotic treatment. Four weeks later, however, the swelling reappeared and was now diagnosed as recurrent Pott's puffy tumor. Interestingly, C-reactive protein levels were in the normal range throughout. Following functional endoscopic frontal sinusotomy, antibiotic treatment with ceftriaxone was administered over 3 weeks and led to complete remission of the lesion. Pott's puffy tumor is a subperiostal abscess of the frontal bone, usually presenting as localized swelling of the soft tissue in the overlying region of the forehead, and is associated with localized osteomyelitis and occasionally with intracranial epidural abscess. The entity has been known since 1768, although recurrent cases with normal inflammation parameters have not been published previously. PMID- 19998015 TI - Bortezomib induces erythema multiforme-like cutaneous adverse effects: report of two cases. PMID- 19998018 TI - [Time delay in resuscitation room treatment and the effect on outcome]. PMID- 19998019 TI - [Fractures of the neck of the femur in younger patients (15-50 years old). Systematic literature research on medial fractures of the neck of the femur in young patients]. AB - Non-pathologic fractures of the neck of the femur in younger patients aged between 15 and 50 years old are rare injuries. These are so-called effectual injuries with very high energy induction due to traffic accidents, falls and sport accidents, causing healthy bones to be fractured and often leading to multiple injuries. The short-term and long-term complications of such injuries sometimes give rise to substantial problems, in particular from non-union fractures and avascular necrosis of the head of the femur. In the literature (and from some experts) the impression is occasionally given that there might be "proven" successful therapeutic procedures for the methods and in particular the timing of treatment of fractures of the neck of the femur in younger patients. This presentation has gone so far that in some cases the term "treatment malpractice" has been used in legal disputes (and judgments) (Judgment IU 5146/00, Higher Regional Court Munich; 2O 861/07 Hei, Regional District Court Ansbach; 118C 421/05, District Court Cologne). The authors have often been called upon as medical experts to comment on the current state of medical knowledge on the question of the biology of healing of fractures of the neck of the femur and the question of"verified" therapy strategies. With this in mind this article is designed to give a review of the current state of proven knowledge according to the available clinical and experimental data and last but not least to stimulate constructive discussion. PMID- 19998020 TI - [Impact of hypothermia on the severely injured patient]. AB - Accidental hypothermia is a common complication in severely injured patients. Risk factors include environmental exposure of the patient at the accident site or in the clinic, infusion of cold fluids, hemorrhagic shock and anesthetics which influence thermoregulation. In contrast to animal studies, human studies and clinical experiences have identified accidental hypothermia of the severely injured patient to be associated with increased complication and mortality rates. As a consequence, hypothermia together with acidosis and coagulopathy, have been coined the lethal triad in severely injured patients. On a cellular level hypothermia reduces cellular activity and metabolism resulting in reduced oxygen consumption, which is therapeutically used in patients following cardiac arrest. However, the activity of important enzymes, such as those of the coagulation pathway, is simultaneously down regulated. Hypothermia-induced coagulopathy, which is refractory to substitution of coagulation factors, is a major complication of hypothermia in traumatized patients. Therefore, hypothermic trauma patients with hemodynamic instability require aggressive rewarming. PMID- 19998021 TI - [Rewarming methods for severely injured hypothermic patients]. PMID- 19998022 TI - [Subtalar dislocation]. AB - Subtalar dislocations represent uncommon injuries of the foot. Leitner [7] described the relationship between medial and lateral dislocations as 6:1. The mechanism is a trauma in plantar flexion/supination of the forefoot with a fixed hindfoot.Immediate reduction, which can usually be performed as a closed reduction is the aim of the treatment. The reduction should be performed under anesthesia in the operating room (OR) and under OR conditions in case the closed reduction shows no success. Subsequently, x-rays and CT scans should be performed in two planes in order to rule out concomitant injuries at the processus posterior tali and the talar head, as bony fragments can necessitate an operative intervention in the case of an interposition of the articulation.In terms of aftercare an immobilization of 6 weeks with a lower leg cast is suggested in the literature. In the presented case an early functional therapy with 2 weeks cast and 4 weeks with a therapy boot could achieve good clinical results. PMID- 19998023 TI - Iliosacral bacterial arthritis and retroperitoneal abscess after tension-free vaginal mesh reconstruction. AB - A 64-year-old woman presented with severe infection accompanied by iliosacral arthritis and retroperitoneal abscess after tension-free vaginal mesh reconstruction using polypropylene meshes (GyneMesh; Gynecare, Ethicon, Somerville, NJ, USA) for pelvic organ prolapsed quantification of stage 3 pelvic organ prolapse. Three weeks after the operation, she complained of high fever (39 degrees C) and sharp pain from the left buttock down to the left lower limb. Computed tomography revealed a retroperitoneal abscess and osteolysis caused by iliosacral arthritis. Because the infection was not resolved by drainage and antibiotic therapy, the left portion of the mesh was removed. She recovered after mesh removal, and no recurrent abscess was observed in the CT after a 6-month follow-up. However, the patient complained of sporadic discomfort in the left buttock, and a blood examination performed during this period, revealed a slight elevation in the C-reactive protein level. Although this symptom was resolved by oral antibiotics, further follow-up was required. PMID- 19998024 TI - Long-term results of vaginal repairs with and without xenograft reinforcement. AB - INTRODUCTION AND HYPOTHESIS: The aim of this paper is to study if xenograft reinforcement of vaginal repair reduces recurrence of prolapse. METHODS: Results 1-5 years after vaginal repair were studied in 41 cases with xenograft and in 82 matched controls without. Symptoms were evaluated by a validated questionnaire and anatomy by pelvic organ prolapse quantification (POPQ). RESULTS: Significant more cases, 97% versus 81% controls, felt cured or much improved (p = 0.02); 11% of cases and 19% of controls had POP symptoms, POPQ > -1 was found in 31% cases and 24% controls. Defining recurrence as POPQ > -1 plus symptoms revealed recurrence in 3% of cases and 12% controls. None of the recurrence rates was significantly different for cases versus controls. No vaginal erosions were seen. Previous surgery was a significant risk factor with odds ratio 7.3 for another recurrence. CONCLUSIONS: Recurrence rates defined by POPQ plus symptoms were low compared to literature. Xenograft reinforcement might improve results. PMID- 19998025 TI - Electrochemical evaluation and determination of antiretroviral drug fosamprenavir using boron-doped diamond and glassy carbon electrodes. AB - Fosamprenavir is a pro-drug of the antiretroviral protease inhibitor amprenavir and is oxidizable at solid electrodes. The anodic oxidation behavior of fosamprenavir was investigated using cyclic and linear sweep voltammetry at boron doped diamond and glassy carbon electrodes. In cyclic voltammetry, depending on pH values, fosamprenavir showed one sharp irreversible oxidation peak or wave depending on the working electrode. The mechanism of the oxidation process was discussed. The voltammetric study of some model compounds allowed elucidation of the possible oxidation mechanism of fosamprenavir. The aim of this study was to determine fosamprenavir levels in pharmaceutical formulations and biological samples by means of electrochemical methods. Using the sharp oxidation response, two voltammetric methods were described for the determination of fosamprenavir by differential pulse and square-wave voltammetry at the boron-doped diamond and glassy carbon electrodes. These two voltammetric techniques are 0.1 M H(2)SO(4) and phosphate buffer at pH 2.0 which allow quantitation over a 4 x 10(-6) to 8 x 10(-5) M range using boron-doped diamond and a 1 x 10(-5) to 1 x 10(-4) M range using glassy carbon electrodes, respectively, in supporting electrolyte. All necessary validation parameters were investigated and calculated. These methods were successfully applied for the analysis of fosamprenavir pharmaceutical dosage forms, human serum and urine samples. The standard addition method was used in biological media using boron-doped diamond electrode. No electroactive interferences from the tablet excipients or endogenous substances from biological material were found. The results were statistically compared with those obtained through an established HPLC-UV technique; no significant differences were found between the voltammetric and HPLC methods. PMID- 19998026 TI - Determination of insoluble avian eggshell matrix proteins. AB - The organic components of bones and other mineralized tissues have a high impact on the organization and deposition of calcium, and consequently influence the mechanical properties of those tissues. The extractable proteins of avian eggshells have been studied extensively and many of them have been identified; insoluble (non-extractable) proteins have been sparsely studied, however. In the work discussed in this paper we studied EDTA-insoluble proteins by gradual decalcification of eggshell with EDTA. The insoluble proteinaceous films were chemically treated with cyanogen bromide and the mixtures of large fragments obtained were gradually precipitated with salt. The separated fractions were digested with trypsin and analyzed by HPLC-MS-MS (ion trap mass spectrometer). Analysis of the entire eggshell matrix (without precipitation steps) only enabled 6 proteins to be determined (ovocalyxins 32 and 36, ovocleidin 17 and 116, clusterin, and ovalbumin). Pretreatment of the individual eggshell layers and gradual precipitation with salt markedly increased the number of proteins identified - 28 proteins were determined. We identified for the first time collagens I (two chains) and III in the eggshell matrix, and Kunitz-like protease inhibitor as a major shell matrix protein. Besides the above mentioned proteins we can also mention EDIL3, fibronectin, sulfhydryl oxidase, tubulin alpha 1, lysozyme, Dickkopf-related protein 3, keratins, and ovotransferrin. The relative abundances of proteins in all eggshell layers were determined using the exponentially modified protein abundance index (emPAI). In the cuticle layer seven proteins were identified, whereas 16 proteins were described in the palisade layer and 23 in the mammillary layer. PMID- 19998027 TI - Highly sensitive rare cell detection based on quantum dot probe fluorescence analysis. AB - This study presents an efficient and sensitive method for detecting rare cells without cell culture, in which cells are analyzed quantitatively using quantum dots (QDs) as a fluorescent probe. By the conjugation of QDs with cells, the biotin-streptavidin reaction functions as a bridge to connect QDs and cells. The cells can be quantified based on the correlation of the QD fluorescence intensity with the cell population. Non-specific adsorption and cross-reaction of QD625 streptavidin on T cell membrane are neglected by reacting with biotin anti-human CD3 and mixing with red blood cell, respectively. Additionally, the photo activation period and pH can be controlled to enhance the fluorescence of cell populations, which increases linearly with the number of T cells from 40 to 100,000, not only in a single T cell line but also in mixing with a total of 10(6) red blood cells. Moreover, the specific T cells can be detected in less than 15 min, even though rare specific cells may number only 40 cells. Among the advantages, the proposed system for detecting rare cells include simplicity of preparation, low cost, rapid detection, and high sensitivity, all of which can facilitate the detection of circulating tumor cells in early stages of diagnosis or prognosis. PMID- 19998028 TI - New trends in fast and high-resolution liquid chromatography: a critical comparison of existing approaches. AB - Recent developments in chromatographic supports and instrumentation for liquid chromatography (LC) are enabling rapid and highly efficient separations. Various analytical strategies have been proposed, for example the use of silica-based monolithic supports, elevated mobile phase temperatures, and columns packed with sub-3 microm superficially porous particles (fused core) or with sub-2 microm porous particles for use in ultra-high-pressure LC (UHPLC). The purpose of this review is to describe and compare these approaches in terms of throughput and resolving power, using kinetic data gathered for compounds with molecular weights ranging between 200 and 1300 g mol(-1) in isocratic and gradient modes. This study demonstrates that the best analytical strategy should be selected on the basis of the analytical problem (e.g., isocratic vs. gradient, throughput vs. efficiency) and the properties of the analyte. UHPLC and fused-core technologies are quite promising for small-molecular-weight compounds, but increasing the mobile phase temperature is useful for larger molecules, for example peptides. PMID- 19998029 TI - A retrospective case-control study assessing the role of trabecular bone score in postmenopausal Caucasian women with osteopenia: analyzing the odds of vertebral fracture. AB - This case-control study assessed whether the trabecular bone score (TBS), determined from gray-level analysis of DXA images, might be of any diagnostic value, either alone or combined with bone mineral density (BMD), in the assessment of vertebral fracture risk among postmenopausal women with osteopenia. Of 243 postmenopausal Caucasian women, 50-80 years old, with BMD T-scores between -1.0 and -2.5, we identified 81 with osteoporosis-related vertebral fractures and compared them with 162 age-matched controls without fractures. Primary outcomes were BMD and TBS. For BMD, each incremental decrease in BMD was associated with an OR = 1.54 (95% CI = 1.17-2.03), and the AUC was 0.614 (0.550-0.676). For TBS, corresponding values were 2.53 (1.82-3.53) and 0.721 (0.660-0.777). The difference in the AUC for TBS vs. BMD was statistically significant (p = 0.020). The OR for (TBS + BMD) was 2.54 (1.86-3.47) and the AUC 0.732 (0.672-0.787). In conclusion, the TBS warrants a closer look to see whether it may be of clinical usefulness in the determination of fracture risk in postmenopausal osteopenic women. PMID- 19998030 TI - MEPE's diverse effects on mineralization. AB - Matrix extracellular phosphoglycoprotein (MEPE) is an inhibitor of mineralization in situ and in cell cultures where altered expression is associated with oncogenic osteomalacia and hypophosphatemic rickets. The purpose of this study was to determine whether the intact protein or the peptide(s) originating from this protein was responsible for the inhibition. The ability of the intact protein and the acidic, serine- and aspartate-rich MEPE-associated motif (ASARM) peptide to promote or inhibit de novo hydroxyapatite formation and growth of hydroxyapatite seed crystals, in both phosphorylated and dephosphorylated forms, was assessed at room temperature in a dynamic gel diffusion system at 3.5 and 5 days. The most effective nucleator concentration was also examined when associated with fibrillar type I collagen. The phosphorylated intact protein was an effective promoter of mineralization in the gelatin gel diffusion system, while the ASARM peptide was an effective inhibitor. When dephosphorylated both the intact protein and the ASARM peptide had no effect on mineralization. Associated with collagen fibrils, some of the effect of the intact protein was lost. This study demonstrates the importance of posttranslational modification for the site-specific activity of MEPE and its ASARM peptide. PMID- 19998031 TI - Magnetic resonance diffusion tensor imaging (MRDTI) and tractography in children with septo-optic dysplasia. AB - BACKGROUND: Septo-optic dysplasia (SOD) refers to a heterogeneous group of midline brain developmental anomalies, with optic nerve hypoplasia (ONH) being one of the morphologic correlates of the condition. Traditionally, ONH has been diagnosed on fundoscopic exam. Conventional MRI is used in cases of suspected ONH to identify associated brain abnormalities and to compare findings to the fundoscopic exam. Advances in magnetic resonance diffusion tensor imaging (MRDTI) permit in vivo, noninvasive, quantitative characterization of the entire visual pathway at 3.0 T. OBJECTIVE: To investigate the feasibility of MRDTI at 3.0 T in children with SOD to evaluate the entire visual pathway. MATERIALS AND METHODS: MRDTI at 3T was performed in two children with SOD and seven age-matched controls. Manual region-of-interest analysis was used to evaluate the tensor metrics of the optic nerves. Deterministic tractography was used to evaluate the tensor metrics of the optic radiations. RESULTS: The SOD patients demonstrated a significant decrease in anisotropy and increase in mean diffusivity of the optic nerves and radiations compared to the control subjects. CONCLUSION: This study demonstrates the feasibility of MRDTI to evaluate the entire visual pathway in children, and it demonstrates pre- and post-chiasmatic diffusion tensor abnormalities in SOD patients. PMID- 19998032 TI - Chronic perilunate dislocations treated with open reduction and internal fixation: results of medium-term follow-up. AB - The purpose of this study was to evaluate the medium-term follow-up results of a series of ten perilunate dislocations treated operatively at least three weeks following injury. The intervals from injury to treatment of the delayed and the chronic groups were four weeks (range, three to six weeks) and 17 weeks (range, 11-25 weeks), respectively. The average follow-up time was 90 months. Surgical procedures included open reduction, internal fixation, grafting, ligament repair, and external fixation. Clinical function was evaluated by the Cooney clinical scoring system. The radiological assessment included the radiolunate angle, scapholunate angle, the revised carpal height ratio, and presence or absence of midcarpal arthritis. In the delayed group, there were two excellent, one good and one poor results. The average postoperative clinical score was 81 (good). In the chronic group, there were four good, one fair, and one poor results. At the last follow-up, the average postoperative clinical score was 76.7 (good). Our results indicate that open reduction and internal fixation can be applied in the treatment of delayed and some of the chronic perilunate dislocations and achieve satisfactory results. PMID- 19998033 TI - Arthroscopic-assisted locking compression plate clavicular hook fixation for unstable fractures of the lateral end of the clavicle: a prospective study. AB - The aim of this prospective study was to assess the clinical outcomes of an unstable fracture of the lateral end of the clavicle treated with an arthroscopic assisted locking compressive plate (LCP) clavicular hook plate. Twenty-three patients underwent arthroscopic assisted LCP clavicular hook plate fixation for these fractures. All patients achieved clinical and radiological union over a mean of 4.2 months (range, 3.4-5 months). Four patients (17%) showed some degree of acromial osteolysis. Three patients (13%) showed radiological signs of arthrosis of the acromioclavicular joint. In one patient, a second fracture (stress) was observed between the medial two screws of the plate without an additional injury. Five patients (22%) showed subacromial bursitis on dynamic ultrasonography. The mean Constant and Murley score was 91 points (range, 81-98). The average level of pain in the shoulder at rest and on abduction was 1 (range, 0-2) and 2.4 (range, 0-4), respectively. Based on our experience, arthroscopic assisted LCP hook plate fixation for the treatment of unstable fractures of the lateral end of the clavicle is not without complications. However, it is an acceptable alternative method that is easy to apply with good results. Furthermore, it prevents rotator cuff impingement, allows early mobilisation and maintains the acromioclavicular joint biomechanics. PMID- 19998034 TI - Sagittal osteotomy of the patella after Morscher. AB - The aim of this study was to present long-term results of sagittal osteotomy of the patella after Morscher. The study included 70 patients treated for patellar dysplasia with a postoperative follow-up from nine to 20 years. There were 59 females and 11 males with an average age of 21 years at the time of operation. Results were evaluated for the entire group of 70 patients and then presented separately for the 30 patients in whom sagittal osteotomy of the patella was performed as a single procedure and 40 patients in whom, in addition to sagittal osteotomy of the patella, medialisation and ventralisation or distalisation of the tibial tuberosity were also performed. The mean value of the Wiberg patellar angle was 112 degrees preoperatively and 140 degrees postoperatively. In the whole group of patients excellent results were obtained in 24 (35%), good results in 30 (42%), fair results in 13 (19%) and poor results in three (4%) patients. In 30 patients with sagittal osteotomy as a single operation excellent results were obtained in 13 (43%), good results in 14 (47%) and fair results in three (10%) patients, while in the 40 patients with sagittal osteotomy and additional surgical procedures, 11 (27%) were excellent, 16 (40%) good, ten (25%) fair and three (8%) had poor results. Sagittal osteotomy of the patella after Morscher is an enrichment of the spectrum of the secondary-causal and preventive procedures for the treatment of patello-femoral disorders. PMID- 19998035 TI - The new "dual osteotomy": combined open wedge and tibial tuberosity anteriorisation osteotomies. AB - The high frequency with which medial compartment osteoarthritis is associated with patellofemoral osteoarthritis makes the addition of tibial tuberosity anteriorisation to high tibial osteotomy an appealing solution, despite the discouraging previously reported long-term results when tubercle anteriorisation was combined with a Coventry closed wedge technique. We conducted a prospective study of a new osteotomy combination: "the dual osteotomy". An open wedge high tibial osteotomy was combined with 1- to 1.5-cm Maquet-like tibial tuberosity anteriorisation. Thirty-four knees in 30 patients underwent surgery, including ten knees in nine male patients and 24 knees in 21 female patients with a mean age of 45 years (age range 34-58 years). All patients had varus medial compartment osteoarthritis and patellofemoral osteoarthritis with preoperative anatomical tibiofemoral angle exceeding 5 degrees . Twenty-four months after surgery, final evaluation detected improvement in the Knee Society clinical rating system function score from a mean of 61.3 (range 30-80) preoperatively to a mean of 87.3 (range 50-100) postoperatively and in the knee pain score from 27.3 (range 10-30) to 47 (range 30-50) postoperatively. Based on the rating system, at final follow-up, 70% of patients experienced no pain, 13% had mild or occasional pain, 10% had pain on stairs only, and 7% had pain during walking and on stairs. Anatomical tibiofemoral angles from 0 to 10 degrees valgus were achieved in 91% of operated knees, and union was achieved in all cases within six to twelve weeks after surgery. The dual osteotomy was effective in the short term in cases of medial compartment osteoarthritis associated with patellofemoral osteoarthritis. PMID- 19998037 TI - Improving cellular properties for genetic manipulation by dispersed growing mutagenesis in Myxococcus fulvus HW-1. AB - One of the key limitations to genetic manipulation in myxobacteria is that the cells grow in clumps in liquid. A salt-tolerant strain HW-1 of Myxococcus fulvus was treated with UV irradiation and produced a completely dispersedly growing mutant UV684. There were no significant differences between the parent HW-1 and the mutant UV684 in terms of salt-tolerant growth. The mutant UV684 and the parent strain had the similar abilities of the fruiting body formation and S motility. Interestingly, the mutant exhibited high transformation/transposition efficiency with 10(5)-10(6) colony-forming units per microg DNA, which was about 10(3)-10(5) fold higher than HW-1. The results indicate that the mutation that led to dispersed growth in the UV684 mutant strain had a few impacts on social behavior, but greatly facilitated molecular genetic manipulation. PMID- 19998038 TI - TvDim1 of Trichoderma virens is involved in redox-processes and confers resistance to oxidative stresses. AB - The evolutionarily conserved Dim1 proteins belong to the TRX fold superfamily. An EST showing high identity values with genes coding for Dim1 proteins was selected from an EST library collection of Trichoderma virens T59. Here, we report the cloning, characterization, and functional analysis of a T. virens T59 TvDim1 gene. The TvDim1 gene, with a sequence size of 614 bp, was PCR-amplified and found to contain three introns. The TvDim1 gene was present as a single copy in the T. virens genome and was also present in another five Trichoderma strains investigated. Increased levels of expression and redox-activity were detected when the fungus was grown in the presence of H(2)O(2). The overexpression and silencing of TvDim1 in T. harzianum T34 gave rise to transformants, with higher and lower growth, redox-activity, and quantities of biomass, respectively, than the wild-type strain after culture under oxidative stress. PMID- 19998039 TI - The complete mitochondrial genome sequence of the hornwort Phaeoceros laevis: retention of many ancient pseudogenes and conservative evolution of mitochondrial genomes in hornworts. AB - Plants have large and complex mitochondrial genomes in comparison to other eukaryotes. In bryophytes, the mitochondrial genomes exhibit a mixed mode of conservative and dynamic evolution. Here, we sequenced the complete mitochondrial genome from hornwort Phaeoceros laevis, to investigate the level of conservation in mitochondrial genome evolution within hornworts. The circular molecule consists of 209,482 base pairs and represents the largest known mitochondrial genome of bryophytes. It contains 30 protein genes, 3 rRNA genes, and 21 tRNA genes, with 34 cis-spliced group II introns disrupting 16 protein genes. There are 11 pseudogenes in this genome, and nine of them are shared with the other fully sequenced hornwort chondriome from Megaceros aenigmaticus, a distant relative of P. laevis. These pseudogenes were likely formed during an early stage of hornwort evolution. The two hornwort chondriomes differ by four inversions and translocations, seven genes, and four introns in the genome structure and organization. At the sequence level, they are very similar, with the identity values ranging mostly from 80 to 95% in intergenic spacers, introns, and exons. These data indicate that mitochondrial genome evolution in hornworts is less conservative than in liverworts, but has not reached the dynamic level as seen in seed plants. PMID- 19998040 TI - Emergency use of an endobronchial one-way valve in the management of severe air leak and massive subcutaneous emphysema. AB - Bronchopleural (BPF) and alveolar-pleural (APF) fistulas are frequently encountered in clinical practice with persistent air leaks that can lead to significant morbidity, prolonged hospital stay, and potentially increased mortality. BPF and APF are commonly related to pulmonary resections. Other etiologies include minimally invasive procedures (thoracentesis and image-guided biopsies), and spontaneous fistulas related to an underlying structural lung disease (e.g., emphysema) or a necrotizing pulmonary process (e.g., infection or malignancy). Radiofrequency ablation for pulmonary malignancies is an effective modality that can rarely lead to APF with persistent air leak. Surgical intervention remains the standard treatment option for BPF and APF. A variety of minimally invasive bronchoscopic approaches can be considered for selected nonsurgical candidates. The use of one-way endobronchial valves to manage severe and persistent air leaks can be considered a minimally invasive option in selected patients. The valves selectively block inspiratory airflow to a specific segmental or subsegmental airway but allow expiratory flow with drainage of air and secretions from the corresponding distal airways and lung parenchyma. PMID- 19998041 TI - Aerosolized polymerized type I collagen reduces airway inflammation and remodelling in a guinea pig model of allergic asthma. AB - Collagen-polyvinylpyrrolidone (Collagen-PVP) has been demonstrated to elicit immunomodulatory properties in different chronic inflammatory diseases. Nevertheless, its effects on asthma are still unknown. We have evaluated whether collagen-PVP could modulate airway inflammation and remodelling in a guinea pig model of allergic asthma. Sensitized guinea pigs were challenged with the allergen (ovalbumin) six times (at 10-day intervals). From the third challenge on, animals were treated every 5 days with saline aerosols containing 0.16, 0.33, or 0.66 mg/ml of collagen-PVP (n = 5, respectively). Some guinea pigs, sensitized and challenged with saline as well as treated with 0 or 0.66 mg/ml collagen-PVP, were included in the study as control (n = 7) and sham groups (n = 5), respectively. From the first challenge on, ovalbumin induced a transient airway obstruction, measured by barometric plethysmography, which was not modified by collagen-PVP treatments. After the last allergen challenge, guinea pigs were anesthetized to obtain bronchoalveolar lavage (BAL) and the left lung caudal lobe. As expected, BAL cell count from allergen-challenged guinea pigs showed abundant neutrophils and eosinophils, as well as numerous tumor necrosis factor (TNF)-alpha-expressing granulocytes and macrophages in airway wall (determined by immunohistochemical assay). Neutrophilia and TNF-alpha-expressing leukocytes, from collagen-PVP treated animals, diminished from 0.16 mg/ml, and eosinophilia from 0.66 mg/ml of collagen-PVP doses. Histological changes induced by allergen challenges include thickening of connective tissue below airway epithelium and vascular wall widening of airway adjacent vessels; these changes were reduced by collagen-PVP treatment. Collagen-PVP seems to have anti-inflammatory and antifibrotic properties in this guinea pig asthma model. PMID- 19998042 TI - Non-colorectal, non-neuroendocrine, and non-sarcoma metastases of the liver: resection as a promising tool in the palliative management. AB - PURPOSE: Hepatic resection is established as a safe procedure for colorectal, neuroendocrine, and sarcoma liver metastases. The present study evaluates whether liver resection is an option for patients with non-colorectal, non neuroendocrine, and non-sarcoma metastases of the liver. METHODS: According to data from our prospective clinical tumor registry, we reviewed the medical records of 44 consecutive patients with non-colorectal, non-neuroendocrine, and non-sarcoma liver metastases, who underwent hepatic resection from January 2000 to December 2008. Univariate Kaplan-Meier analysis and a stepwise multivariable Cox regression model were applied. RESULTS: Following hepatic resection, mean overall survival was 21 months, and 5-year survival was 20%. Following hepatic resection, gender, histology, and chemotherapy were of prognostic value in our patient cohort in univariate analysis (p < 0.05). Multivariate survival analysis confirmed chemotherapy (p = 0.002) as an independent prognostic variable. Following initial resection of the primary tumor, synchrone occurrence of metastases, histology, localization of primary, perioperative complications, interval between initial resection of the primary tumor and resection of the metastases, and metastases in follow-up after hepatic resection were of prognostic value in univariate analysis (p < 0.05). Histology (p = 0.017) and interval between resection of the primary and resection of the metastases (p = 0.030) were confirmed as independent prognostic variables in multivariate survival analysis. CONCLUSIONS: Hepatic resection seems to be a safe and promising additive for a selective group of patients with non-colorectal, non neuroendocrine, and non-sarcoma metastases of the liver. PMID- 19998043 TI - Pediatric community-acquired pneumonia in the ambulatory setting. PMID- 19998044 TI - Hepatitis-associated aplastic anemia during a primary infection of genotype 1a torque teno virus. AB - A 12-year-old Japanese boy suffered from severe acute hepatitis and pancytopenia. The patient underwent successful bone marrow transplantation from an HLA identical sister. Torque teno virus (TTV) DNA of genotype 1a and IgM-class antibody against the virus were detected in sera at the onset of hepatitis. TTV/1a DNA and anti-TTV/1a IgM antibody levels were undetectable on the 16th and 46th days after the onset of illness, respectively. Anti-TTV/1a IgG antibody was positive throughout the observation period. Sequential viral load and anti-TTV/1a IgM antibody suggested a primary infection of TTV/1a. Genomic sequence of the virus coincided with that of the original strain first isolated from human. TTV DNA was quantified at 130 copies in 10(5) bone marrow mononuclear cells, which suggested that infection of hematopoietic cells might be the cause of aplasia. This is the first report of TTV hepatitis-associated aplastic anemia assessed by the anti-TTV antibodies and viral load in peripheral blood and bone marrow. PMID- 19998045 TI - Living with multiple myeloma: experiences of patients and their informal caregivers. AB - GOALS OF WORK: The purpose of this study was to explore the experience of living with myeloma after completion of treatments in patients and their informal caregivers. PATIENTS AND METHODS: Qualitative interviews were carried out with 20 patients living with myeloma and 16 of their informal caregivers (mostly spouses), asking them through semi-structured interviews to talk about the effects of myeloma on their lives, issues and concerns, their supportive care needs and how they were coping in everyday life. MAIN RESULTS: Key findings indicate the significant impact myeloma has on patients' and caregivers' emotional, role, social and work-related areas of life. While patients seemed less engaged with their illness, their informal caregivers were providing practical and emotional support to patients almost exclusively, often by neglecting their own needs. This increased engagement with the informal caring often led to experiences of a heightened illness burden and difficulties with coping. Both patients and caregivers had significant fears and uncertainty about the future, with myeloma being described as a 'time bomb'. Both patients and caregivers were concealing stressful situations related to the illness from each other in an effort to protect them, although this resulted in isolation. CONCLUSIONS: While health professionals' attention has been concentrated in helping patients to get through treatments, it is imperative that psychosocial rehabilitation is offered also to those who survive many years after diagnosis and are off treatments. Caregivers are particularly vulnerable to the high demands of caring for myeloma patients, and a more concerted action by health professionals should be directed to them. PMID- 19998046 TI - Radiodermatitis prevention with sucralfate in breast cancer: fundamental and clinical studies. AB - BACKGROUND: Acute radiodermatitis induced by radiotherapy may affect the quality of life and in some cases requires withholding treatment. The present study concerns the protective effect of a 1% sucralfate lotion. We propose joint fundamental and clinical points of view. METHODS: The free radical scavenging capacity of sucralfate was measured with electron spin resonance and was supported by theoretical calculations. The clinical effects of sucralfate lotion were evaluated on 21 women treated for breast cancer. Breast skin response was evaluated at 0, 10, 20, 30, 40, and 50 Gy, according to (1) the radiation therapy oncology group (RTOG) acute toxicity scale and (2) spectrophotometry data obtained with X-Rite SP60. RESULTS AND CONCLUSIONS: Sucralfate appeared as a relatively poor free radical scavenger (compared to reference compounds such as vitamin E). The sucralfate-containing lotion used in the present study did not provide systematic radiodermatitis prevention. Spectrophotometric evaluation of the skin response to irradiation appeared to be a very effective and more sensitive technique than the RTOG scale. Its use should be recommended to study cutaneous radioprotective action. PMID- 19998047 TI - Subtype identification of the novel A H1N1 and other human influenza A viruses using an oligonucleotide microarray. AB - A novel strain of influenza A (H1N1) virus was isolated in Mexico and the US in March and April 2009. This novel virus spread to many countries and regions in a few months, and WHO raised the level of pandemic alert from phase 5 to phase 6 on June 11, 2009. The accurate identification of H1N1 virus and other human seasonal influenza A viruses is very important for further treatment and control of their infections. In this study, we developed an oligonucleotide microarray to subtype human H1N1, H3N2 and H5N1 influenza viruses, which could distinguish the novel H1N1 from human seasonal H1N1 influenza viruses and swine H1N1 influenza viruses. The microarray utilizes a panel of primers for multiplex PCR amplification of the hemagglutinin (HA), neuraminidase (NA) and matrix (MP) genes of human influenza A viruses. The 59-mer oligonucleotides were designed to distinguish different subtypes of human influenza A viruses. With this microarray, we accurately identified and correctly subtyped the reference virus strains. Moreover, we confirmed 4 out of 39 clinical throat swab specimens from suspected cases of novel H1N1. PMID- 19998048 TI - Do drainage liquid characteristics serve as predictors for seroma formation after incisional hernia repair? AB - BACKGROUND: Though the occurrence of postoperative seroma after incisional hernia repair using mesh reinforcement is very common, little is known about the genesis of seroma formation. The aim of this study was to determine the characteristics of drainage liquid as a potential predictor for the development of seroma after incisional hernia mesh repair. Furthermore, the characteristics of drainage liquid were compared to the characteristics of seroma liquid. METHODS: The incidence of postoperative seroma associated with pH value, concentration of lactate, total protein, albumin, propeptide-III-procollagen (P-III-P), hyaluronan, fibronectin and IL-1 receptor antagonist (IL-1-RA) in the drainage liquid were prospectively determined in 38 patients who underwent incisional hernia repair by lightweight polypropylene-polyglactin composite mesh (Vypro-II). The findings were compared to the seroma liquid characteristics of those patients who developed a seroma formation. RESULTS: In 11 patients (29%), seroma formation was present after removal of the drainage. We observed significantly elevated mean drainage volume (148 +/- 64 ml vs. 93 +/- 71 ml; P = 0.014) and significantly reduced pH value (7.26 +/- 0.12 vs. 7.41 +/- 0.23; P = 0.016) and IL-1-RA (100 +/- 71 U/ml vs. 145 +/- 108 U/ml; P = 0.016) in the drainage liquid of patients with seroma formation in comparison to patients without seroma formation. In addition, we found significantly altered concentration of lactate (9.8 +/- 2.0 mmol/l vs. 5.5 +/- 1.4 mmol/l; P < 0.001), P-III-P (24 +/- 13 U/ml vs. 89 +/- 79 U/ml; P = 0.045) and fibronectin (0.10 +/- 0.03 g/l vs. 0.24 +/- 0.13 g/l; P = 0.005) in the drainage liquid in comparison to seroma liquid. CONCLUSIONS: The pH value of wound solution proves to be a reliable predictor for the subsequent presence of seroma formation. Furthermore, our findings implicate that seroma formation cannot be seen as persistent drainage liquid. PMID- 19998049 TI - Dynamics of point defects and stripe textures in Smectic-C Langmuir monolayers. AB - Langmuir monolayers of an azobenzene fatty acid derivative have been studied experimentally in a regime where confined domains with Smectic-C order form spontaneously. Coalescence of domains results in a dynamics of formation and annihilation of point defects and string-like distortions of the molecular field amenable to semi-quantitative analysis. Absence of backflow and layer thickness effects enables us to extract values for material parameters from the analysis of defect dynamics. PMID- 19998050 TI - Frequency-dependent deformation of liquid crystal droplets in an external electric field. AB - Nematic droplets suspended in the isotropic phase of the same substance were subjected to alternating electrical fields of varying frequency. To keep the system at a constant nematic/isotropic volume ratio with constant droplet size, we carefully kept the temperature in the isotropic/nematic coexistence region, which was broadened by adding small amounts of a non-mesogenic liquid. Whereas the nematic droplets remained spherical at low (in the order of 10 Hz) and high frequencies (in the order of 1 kHz), at intermediate frequencies we observed a marked flattening of the droplets in the plane perpendicular to the applied field. Droplet deformation occurred both in liquid crystals (LCs) with positive and negative dielectric anisotropy. The experimental data can be quantitatively modelled with a combination of the leaky dielectric model and screening of the applied electric field due to finite conductivity. PMID- 19998051 TI - Rheology of a sonofluidized granular packing. AB - We report experimental measurements on the rheology of a dry granular material under a weak level of vibration generated by sound injection. First, we measure the drag force exerted on a wire moving in the bulk. We show that when the driving vibration energy is increased, the effective rheology changes drastically: going from a non-linear dynamical friction behavior --weakly increasing with the velocity-- up to a linear force-velocity regime. We present a simple heuristic model to account for the vanishing of the stress dynamical threshold at a finite vibration intensity and the onset of a linear force velocity behavior. Second, we measure the drag force on spherical intruders when the dragging velocity, the vibration energy, and the diameters are varied. We evidence a so-called "geometrical hardening" effect for smaller-size intruders and a logarithmic hardening effect for the velocity dependence. We show that this last effect is only weakly dependent on the vibration intensity. PMID- 19998052 TI - Necrotizing pneumonia in Greece caused by a USA400 (ST1) Staphylococcus aureus harboring SSCmec type V. PMID- 19998053 TI - Urinary tract infections caused by multi-drug resistant Proteus mirabilis: Risk factors and clinical outcomes. AB - BACKGROUND: Proteus mirabilis (PM) as well as other members of the Enterobacteriaceae family are a leading cause of infectious diseases in both the community and acute care settings. The prevalence of multi-drug resistant (MDR) bacterial isolates have increased in the last few years, affecting the prognosis and survival of hospitalized patients. The aim of our study was to determine the risk factors and clinical outcomes of urinary tract infections (UTIs) caused by MDR PM in patients hospitalized in our institution. METHODS: This was a retrospective matched case-control study. Records of patients with PM-positive urine culture were reviewed, and data were included for analysis. RESULTS: Univariate analysis revealed that the variables significantly associated with acquisition of MDR PM vs non-MDR PM UTI were younger age ([in years] median 77.5, range 20-94 vs median 78, range 40-94, p = 0.04), other concomitant infectious diseases (57.1 vs 35.7%, p = 0.037),number of prior infectious diseases (mean 0.95 +/- 0.99 vs 0.57 +/- 0.85, p = 0.035), diagnosis of infection at hospital admission (67.9 vs 42.9%, p = 0.008), and prior therapy with antipseudomonal penicillin (17.9 vs 1.8%, p = 0.004),respectively. Mean length of hospitalization was 29.95 days for the MDR group and 30.04 days for the non-MDR group(p = non significant [NS]). The crude mortality rate following hospital admission was 19/56 (33.9%) vs 14 (25%)in the MDR PM and non-MDR PM groups, respectively(p = 0.300, odds ratio [OR] 1.54, 95% confidence interval[CI] 0.63-3.82). The production of extended-spectrum beta lactamases(ESBL) was found in 100% of MDR-PM vs 31.5%of non-MDR-PM urine isolates (p < 0.001). All variables found to be significantly associated with MDR-PM UTI were included in a logistic regression model. Independent risk factors for MDR-PM UTI were empiric cephalosporin therapy(OR 4.694, 95% CI 1.76-12.516, p = 0.002) and prior antipseudomonal penicillin (piperacillin/tazobactam) therapy during the last year (OR 11.175, 95% CI 1.09-114.2,p = 0.04). CONCLUSIONS: Prior piperacillin/tazobactam and empiric cephalosporin use were the independent risk factors of MDR-PM strains. All MDR-PM urinary isolates at our institution were ESBL producers. Therefore, carbapenem use remains the only available treatment option for MDR-PM isolates in our institution. PMID- 19998054 TI - Choroidal granuloma as an initial manifestation of systemic sarcoidosis. AB - Systemic sarcoidosis presenting as a choroidal nodule is a rare phenomenon, with limited reports in the past. We present a case of systemic sarcoidosis in a young adult male presenting as a solitary choroidal mass. The diagnosis was made based on elevated serum lysozyme, serum angiotensin-converting enzyme, negative Mantoux, and computed tomography (CT) scan of the chest. Ultrasound examination and fundus fluorescein angiography helped us to differentiate this lesion from choroidal melanoma. Magnetic resonance imaging (MRI) of brain was done to rule out neurosarcoidosis. Patient responded well to systemic steroid therapy, with marked resolution of the choroidal granuloma, and reduced size of mediastinal lymph nodes on repeat CT scan of the chest. PMID- 19998055 TI - Thermosensitive drug delivery system of salmon calcitonin: in vitro release, in vivo absorption, bioactivity and therapeutic efficacies. AB - PURPOSE: The purpose of this study was to develop a biodegradable triblock copolymer, mPEG-PLGA-mPEG-based delivery system for long-term controlled release of salmon calcitonin (sCT) after single subcutaneous injection. METHODS: The delivery system was prepared by dissolving sCT into polymer solution. In vitro release of sCT from the delivery systems was studied in phosphate buffer saline (PBS, pH 7.4) at 37 degrees C. Stability of released sCT and sCT remaining in gel formulation was evaluated using circular dichroism, HPLC and MALDI-TOF mass spectrometry. In vivo absorption and therapeutic efficacy of sCT from the polymeric formulations were examined in female wistar rats and methylprednisolone acetate (MPA)-induced osteoporosis rat model, respectively. RESULTS: The polymeric formulations of sCT showed long term controlled release (~20 to 40 days) of sCT in its conformationally and chemically stable form. The sCT polymeric formulations controlled the release of sCT over ~20 to 40 days and prevented MPA induced osteoporosis in vivo. The released sCT was biologically active in terms of lowering serum calcium level. CONCLUSIONS: The triblock copolymer delivery systems controlled the release of sCT in vitro and in vivo in chemically and conformationally stable as well as biologically active and therapeutically effective form. PMID- 19998056 TI - Pseudovirions as vehicles for the delivery of siRNA. AB - Over the last two decades, small interfering RNA (siRNA)-mediated gene silencing has quickly become one of the most powerful techniques used to study gene function in vitro and a promising area for new therapeutics. Delivery remains a significant impediment to realizing the therapeutic potential of siRNA, a problem that is also tied to immunogenicity and toxicity. Numerous delivery vehicles have been developed, including some that can be categorized as pseudovirions: these are vectors that are directly derived from viruses but whose viral coding sequences have been eliminated, preventing their classification as viral vectors. Characteristics of the pseudovirions discussed in this review, namely phagemids, HSV amplicons, SV40 in vitro-packaged vectors, influenza virosomes, and HVJ Envelope vectors, make them attractive for the delivery of siRNA-based therapeutics. Pseudovirions were shown to deliver siRNA effector molecules and bring about RNA interference (RNAi) in various cell types in vitro, and in vivo using immune-deficient and immune-competent mouse models. Levels of silencing were not always determined directly, but the duration of siRNA-induced knockdown lasted at least 3 days. We present examples of the use of pseudovirions for the delivery of synthetic siRNA as well as the delivery and expression of DNA directed siRNA. PMID- 19998057 TI - Montelukast inhibits matrix metalloproteinases expression in atherosclerotic rabbits. AB - BACKGROUND: Matrix metalloproteinases (MMPs) play important roles in the development and destabilization of atherosclerotic plaques. It is known that montelukast inhibits neointimal hyperplasia. However, the underlying mechanisms for the inhibitory effects of montelukast on neointimal formation have been poorly defined. METHODS: Thirty-six male New Zealand White rabbits were randomized as normal control, placebo (0.9% NaCl, 1.5 ml/kg/day, via intraperitoneal injection), atorvastatin (atorvastatin, 1.5 mg/kg/day, orally) and montelukast groups (montelukast, 1.5 mg/kg/day, via intraperitoneal injection). Atherosclerosis was induced by balloon-injury and high-cholesterol (HC) diet. Serum lipids were measured at 0, 8 and 12 weeks. After 12 weeks, the rabbits were sacrificed and histopathological changes examined. Immunohistochemistry and reverse transcription-polymerase chain reaction were used to measure the expression of MMP-2 and MMP-9 in the plaques. RESULTS: It was found that montelukast reduced neointimal formation, decreased macrophage accumulation, and increased smooth muscle cells. It also attenuated the expression of MMP-2 and MMP-9 in atherosclerotic plaques, but it had no effect on plasma lipid levels. CONCLUSION: These data indicate that montelukast inhibits neointimal hyperplasia in association with decreased expression of MMP-2 and MMP 9 independent of plasma lipid levels in atherosclerotic plaques after vascular injury in hyperlipidemic rabbits. PMID- 19998058 TI - CFD simulation of an internal spin-filter: evidence of lateral migration and exchange flow through the mesh. AB - In the present work Computational Fluid Dynamics (CFD) was used to study the flow field and particle dynamics in an internal spin-filter (SF) bioreactor system. Evidence of a radial exchange flow through the filter mesh was detected, with a magnitude up to 130-fold higher than the perfusion flow, thus significantly contributing to radial drag. The exchange flow magnitude was significantly influenced by the filter rotation rate, but not by the perfusion flow, within the ranges evaluated. Previous reports had only given indirect evidences of this exchange flow phenomenon in spin-filters, but the current simulations were able to quantify and explain it. Flow pattern inside the spin-filter bioreactor resembled a typical Taylor-Couette flow, with vortices being formed in the annular gap and eventually penetrating the internal volume of the filter, thus being the probable reason for the significant exchange flow observed. The simulations also showed that cells become depleted in the vicinity of the mesh due to lateral particle migration. Cell concentration near the filter was approximately 50% of the bulk concentration, explaining why cell separation achieved in SFs is not solely due to size exclusion. The results presented indicate the power of CFD techniques to study and better understand spin-filter systems, aiming at the establishment of effective design, operation and scale-up criteria. PMID- 19998059 TI - Biallelic MYH germline mutations as cause of Muir-Torre syndrome. AB - Muir-Torre syndrome is a rare, inherited disease predisposing of gastrointestinal and cutaneous tumours, such as keratoacanthomas and sebaceous gland adenomas. Muir-Torre syndrome is usually inherited in an autosomal dominant fashion and associated with mutations in the mismatch repair genes, predominantly in MLH1 and MSH2 genes. This report describes a man who has multiple adenomatous colon polyps, a gastric cancer, multiple colorectal cancers and sebaceous adenomas caused by biallelic MYH germline mutations. This finding demonstrates that MYH gene analysis should be considered in Muir-Torre families where no mismatch repair gene mutations have been found. Furthermore, this report contributes to characterize the clinical phenotype caused by biallelic mutations in MYH gene, which may share with other hereditary colon cancer syndromes. PMID- 19998060 TI - Cultures of human tracheal gland cells of mucous or serous phenotype. AB - There are two main epithelial cell types in the secretory tubules of mammalian glands: serous and mucous. The former is believed to secrete predominantly water and antimicrobials, the latter mucins. Primary cultures of human airway gland epithelium have been available for almost 20 yr, but they are poorly differentiated and lack clear features of either serous or mucous cells. In this study, by varying growth supports and media, we have produced cultures from human airway glands that in terms of their ultrastructure and secretory products resemble either mucous or serous cells. Of four types of porous-bottomed insert tested, polycarbonate filters (Transwells) most strongly promoted the mucous phenotype. Coupled with the addition of epidermal growth factor (EGF), this growth support produced "mucous" cells that contained the large electron-lucent granules characteristic of native mucous cells, but lacked the small electron dense granules characteristic of serous cells. Furthermore, they showed high levels of mucin secretion and low levels of release of lactoferrin and lysozyme (markers of native serous cells). By contrast, growth on polyethylene terephthalate filters (Cyclopore) in medium lacking EGF produced "serous" cells in which small electron-dense granules replaced the electron-lucent ones, and the cells had high levels of lactoferrin and lysozyme but low levels of mucins. Measurements of transepithelial resistance and short-circuit current showed that both "serous" and "mucous" cell cultures possessed tight junctions, had become polarized, and were actively secreting Cl. PMID- 19998061 TI - Gene expression profiling of the developing mouse kidney and embryo. AB - The metanephros is formed from the reciprocal inductive interaction of two precursor tissues, the metanephric mesenchyme (MM) and the ureteric bud (UB). The UB induces MM to condense and differentiate forming the glomerulus and renal tubules, whilst the MM induces the UB to differentiate into the collecting tubules of the mature nephron. Uninduced MM is considered the progenitor cell population of the developing metanephros because of its potential to differentiate into more renal cell types than the UB. Previous studies have identified the phenotype of renal precursor cells; however, expression of candidate marker genes have not been analysed in other tissues of the murine embryo. We have assayed up to 19 candidate genes in eight embryonic tissues at five gestation stages of the mouse embryo to identify markers definitively expressed by renal cells during metanephric induction and markers developmentally regulated during kidney maturation. We then analysed their expression in other developing tissues. Results show Dcn, Hoxc9, Mest, Wt1 and Ywhaq were expressed at moderate to high levels during the window of metanephric specification and early differentiation (E10.5-E12.5 dpc), and Hoxc9, Ren1 and Wt1 expression was characteristic of mature renal cells. We demonstrated Cd24a, Cdh11, Mest, Scd2 and Sim2 were regulated during brain development, and Scd2, Cd24a and Sip1 expression was enriched in developing liver. These markers may be useful negative markers of kidney development. Use of a combination of highly expressed and negative markers may aid in the identification and removal of non-renal cells from heterogeneous populations of differentiating stem cells. PMID- 19998062 TI - Acid-base imbalance in uncomplicated ST-elevation myocardial infarction: the clinical role of tissue acidosis. AB - Little information is available on acid-base imbalance in uncomplicated ST elevation myocardial infarction (STEMI) submitted to primary percutaneous intervention (PCI). We therefore assessed acid-base imbalance in 257 consecutive uncomplicated STEMI patients submitted to PCI to determine whether its evaluation could help in identifying patients at higher risk for in-hospital complications (acute pulmonary edema and dysrhythmias). A basic metabolic profile was performed at hospital admission, that is before PCI. After PCI, we measured: creatinine, uric acid and NT-pro BNP and serum electrolytes. Peak troponin I was also considered. Acidemia was present in 11 patients (4.2%), HCO(3) < 22 in 62 (24.1%). Base excess < -3 was detectable in 70 patients (27.2%), anion gap > 12 in 13 (5.1%), Cl/Na < 0.79 in 93 patients (38.5%). Patients with a Cl/Na < 0.79 had a lower LVEF (p = 0.042) and higher values of NT-pro-BNP (p = 0.019) and of latency (p = 0.029) together with a higher length of stay (p = 0.017) and a higher incidence of in-hospital complications (p = 0.017). At backward stepwise regression analysis, the following variables resulted independent predictors of in-hospital complications: base excess OR 1.47 (95% CI 1.04-2.10) p = 0.031; Cl/Na ratio O.R. 1.85 (95% CI 1.05-3.27) p = 0.035. In STEMI patients submitted to mechanical revascularization the evaluation of acid-base status and, in particular the detection of even mild degrees of acidosis may help in risk stratification for in-hospital complications. A Cl/Na < 0.79 ratio and a base excess are independent predictors for in-hospital complications. PMID- 19998063 TI - Visceral obesity and metabolic syndrome: two faces of the same medal? AB - In this review, we have analyzed the role of visceral obesity in the occurrence of metabolic syndrome (MetS). MetS is a common metabolic disorder that has been related recently to the increasing prevalence of obesity. The disorder is defined in various ways, but in the near future a new definition(s) should be applicable worldwide. The pathophysiology has been largely attributed, in the past years, to insulin resistance, although several epidemiological and pathophysiological data now indicate visceral obesity as a main factor in the occurrence of all the components of MetS. In view of this, relationships among visceral obesity, free fatty acids, dyslipidemia and insulin resistance have been reported. In addition, the effects of some adipocytokines and other proinflammatory factors produced by fat accumulation on the occurrence of MetS have been also emphasized. Accordingly, the "hypoadiponectinemia hypothesis" has been proposed as the most interesting to explain the pathophysiology of MetS. The epidemiologic, pathophysiologic and clinical data reported seem to indicate that MetS might be considered a fatal consequence of visceral obesity. PMID- 19998064 TI - Persistence of derivative chromosome 22 after achieving a major molecular response in chronic myeloid leukemia with a cryptic BCR-ABL1 fusion gene. AB - We herein report the findings of a 47-year-old Japanese female with chronic myeloid leukemia (CML) with a cryptic BCR-ABL1 transcript on chromosome 9 and a derivative chromosome 22 unrelated to BCR-ABL1. Although she achieved and continued to demonstrate a major molecular response to imatinib treatment following interferon-alpha, there was persistence of a derivative chromosome 22. A detailed chromosome/molecular studies, including serial karyotyping analysis, finally resulted in the karyotyping at the disease onset to be 47,XX,+del(22)(q11.2), with two genetic evens, namely a cryptic BCR-ABL1 transcript on chromosome 9 and derivative chromosome 22 unrelated to BCR-ABL1. This CML case with these two rare genetic events thus raises diagnostic issues such as the difficulty in making a concise evaluation of the chromosomal/molecular events and an accurate disease prognosis, as well as the difficulty in determining the disease remission status after treatment. PMID- 19998065 TI - Interleukin-2 and granulocyte-macrophage-colony-stimulating factor immunomodulation with high-dose chemotherapy and autologous hematopoietic stem cell transplantation for patients with metastatic breast cancer. AB - Immunomodulation with cytokines was used to improve the result of high-dose chemotherapy (HDC)/autologous hematopoietic stem cell transplantation (AHST). We examined the use of IL-2 and growth factors for mobilization, ex vivo activation of peripheral blood stem cell (PBSC) and maintenance therapy after HDC/AHST in metastatic breast cancer. Eligible patients with metastatic breast cancer for HDC/AHST were assigned to 1 of 3 protocols for PBSC mobilization: G-CSF (group 1); IL-2 + G-CSF (group 2); or IL-2 + G-CSF + GM-CSF (group 3). HDC with cyclophosphamide, carmustine and thiotepa was given from day -7 to -5. PBSCs were treated ex vivo with IL-2 for 24 h and reinfused on day 0. Maintenance therapy included low-dose IL-2, followed by 2 courses of intermediate-dose IL-2. GM-CSF was given from day 1 until neutrophil recovery. Thirty-four patients (10 in group 1, 14 in group 2, and 10 in group 3) were included. Comparable numbers of CD34(+) cells were collected from all 3 groups; incremental increases of CD3(+) cells were collected from groups 1 to 2 and to 3 (p = 0.03). Major adverse effects from IL-2 were fever, hypotension and fatigue; no treatment-related mortality was seen. At a median follow-up of 790.5 days (range 150-2,722 days), median progression-free survival was 434 days and median overall survival was 1,432 days. Estimated 3-year progression-free and overall survival rates were 31 and 57%. Our study suggested that the use of IL-2 and growth factors immunomodulation with HDC/AHST was feasible with comparable survival rates. PMID- 19998066 TI - Development and validation of the pregnancy and infant orientation questionnaire. AB - The purpose of this study was the construction, scaling, and scale validation of a self-report questionnaire assessing biographical information and motivation for pregnancy and infant care in men and women with disorders of sex development or other gender variations of potential clinical relevance. The overall design of the questionnaire and the initial item pool were derived from related clinical and research experience. Collection of pilot data and, where appropriate, scale construction (via principal components analyses) were based on Canadian convenience samples of heterosexual (HET) and non-HET men and women (N = 414). A sample of gender-dysphoric (GDYS) men and women (n = 45) was added for validity analysis. Validation of the resulting scales was based on the demonstration of expected scale differences between HET men and women, as well as between HET, non HET, and GDYS men and between HET, non-HET, and GDYS women, and was successful with one exception. This study concludes that this new questionnaire, in addition to its descriptive sections, provides several distinct scales related to desires for child bearing and child rearing and has good psychometric properties. PMID- 19998067 TI - Evidence that isopropylthioxanthone (ITX) is devoid of anxiolytic and sedative effect. AB - Isopropylthioxanthone (ITX) is a well-known photo-initiator in ultraviolet light cured inks frequently used in milk packaging materials, yoghurt, ready-to-feed infant formula, and other drinks. Traces of ITX have been found in milk and, as a consequence, there was considerable interest in studying the biological activity of this molecule and its potential hazard for the human health. Although the ITX genotoxic effects have been excluded by the European Food Safety Authority (EFSA), the US Environmental Protection Agency (USEPA) is still examining its possible toxic potential depending on a dose-effect ratio. Little is known about the ITX activity on the function of the central nervous system and cerebral neurotransmitters. Using behavioural, biochemical, and electrophysiological tests, the authors have found that: (1) ITX did not exert an in vivo anxiolytic or sedative effect when administered orally to rats; (2) ITX did not affect the binding characteristics of central and peripheral benzodiazepine receptors studied in vitro; and (3) ITX did not influence the ability of gamma-Aminobutyric acid (GABA) to increase the chloride channel permeability studied by patch clamp technique in a single neuron of cultured cerebellar granule cells. PMID- 19998068 TI - Is drowning a serious public health problem in Guangdong Province, People's Republic of China?--results from a retrospective population-based survey, 2004 2005. AB - The aim of this study is to describe the characteristics of fatal drowning in Guangdong Province during the period 2004-2005. Twelve areas were randomly chosen by stratified cluster sampling, and a retrospective cross-sectional survey was conducted in 2006 to collect death cases during the 2004-2005 period. Causes of death relating to drowning were coded according to International Classification of Diseases, the tenth version. During 2004-2005, 849 unintentional drowning deaths occurred, representing a drowning rate of 5.6/100,000 per year. The rate was higher in rural areas (6.4/100,000) than in city (3.7/100,000), and the rate ratio of male to female was 1.5. The elderly aged 80 years and over and children aged 5-9 years had higher mortalities among all age groups, representing rates of 28.8 and 11.0 per 100,000 person years. For children aged 5-9 years, deaths from drowning accounted for the highest proportion (62.5%) of all injury deaths. The most common site of drowning was natural bodies of water (60.8%), and summer was the highest risk season for drowning. The results suggest drowning is an important public health problem, and it primarily threatens children in Guangdong province. Drowning intervention programs should be a health priority in Guangdong province. PMID- 19998069 TI - Estimating the effect of word predictability on eye movements in Chinese reading using latent semantic analysis and transitional probability. AB - Latent semantic analysis (LSA) and transitional probability (TP), two computational methods used to reflect lexical semantic representation from large text corpora, were employed to examine the effects of word predictability on Chinese reading. Participants' eye movements were monitored, and the influence of word complexity (number of strokes), word frequency, and word predictability on different eye movement measures (first-fixation duration, gaze duration, and total time) were examined. We found influences of TP on first-fixation duration and gaze duration and of LSA on total time. The results suggest that TP reflects an early stage of lexical processing while LSA reflects a later stage. PMID- 19998070 TI - The word class effect in the picture-word interference paradigm. AB - The word class effect in the picture-word interference paradigm is a highly influential finding that has provided some of the most compelling support for word class constraints on lexical selection. However, methodological concerns called for a replication of the most convincing of those effects. Experiment 1 was a direct replication of Pechmann and Zerbst (2002; Experiment 4). Participants named pictures of objects in the context of noun and adverb distractors. Naming took place in bare noun and sentence frame contexts. A word class effect emerged in both bare noun and sentence frame naming conditions, suggesting a semantic origin of the effect. In Experiment 2, participants named objects in the context of noun and verb distractors whose word class relationship to the target and imageability were orthogonally manipulated. As before, naming took place in bare noun and sentence frame naming contexts. In both naming contexts, distractor imageability but not word class affected picture naming latencies. These findings confirm the sensitivity of the picture-word interference paradigm to distractor imageability and suggest the paradigm is not sensitive to distractor word class. The results undermine the use of the word class effect in the picture-word interference paradigm as supportive of word class constraints during lexical selection. PMID- 19998071 TI - Response to open peer commentaries on "the coming era of nanomedicine". PMID- 19998072 TI - The era of nanomedicine and nanoethics: has it come, is it still coming, or will it pass us by? PMID- 19998073 TI - Medical care for terrorists--yes to treat! PMID- 19998074 TI - The coming era of nanomedicine. AB - This essay presents some general background on nanomedicine, particularly focusing on some of the investment that is being made in this emerging field. The bulk of the essay, however, consists of explorations of two areas in which the impacts of nanomedicine are likely to be most significant: diagnostics and medical records and treatment, including surgery and drug delivery. Each discussion includes a survey some of the ethical and social issues that are likely to arise in these applications. PMID- 19998075 TI - Nanotechnology will change more than just one thing. PMID- 19998076 TI - Nanomedicine: ethical concerns beyond diagnostics, drugs, and techniques. PMID- 19998077 TI - Nanomedicine and nanomedical ethics. PMID- 19998078 TI - Missing the boat on nanoethics. PMID- 19998079 TI - Legalism, countertransference, and clinical moral perception. AB - This target article focuses on dynamics that arise in three typical ethically complex cases in which psychiatric consultations are requested by physicians: a dying patient refuses life-prolonging treatment, an uncooperative patient demands to be allowed to go outside and smoke, and an angry patient demands to be admitted to the hospital. The discussion canvasses what is at stake morally and clinically in each of these cases and explores clinician-patient interactions, dynamics in relationships between consulting physicians and consultant psychiatrists, patient transference, and physician countertransference. The article defines legalism and countertransference and argues that an ethically and clinically important consequence of these phenomena for patients is distortion of clinicians' perceptions of patients' decisions and vulnerabilities. The discussion also describes how legalism and countertransference adversely affect how clinicians treat their psychiatrist colleagues. Finally, the article suggests how the effects of legalism and countertransference can be mitigated. PMID- 19998080 TI - What reason can do for clinical moral perception. PMID- 19998081 TI - Let's blame the physicians ... again: physician legalism and countertransference. PMID- 19998082 TI - The physician's fragility. PMID- 19998083 TI - Some distinctions, "hair splitting," and added worries. PMID- 19998084 TI - Do all physicians need to recognize countertransference? PMID- 19998085 TI - Medical care for terrorists--to treat or not to treat? AB - With the escalation of terrorism worldwide in recent years, situations arise in which the perpetration of violence and the defense of human rights come into conflict, creating serious ethical problems. The Geneva Convention provides guidelines for the medical treatment of enemy wounded and sick, as well as prisoners of war. However, there are no comparable provisions for the treatment of terrorists, who can be termed unlawful combatants or unprivileged belligerents. Two cases of severely injured terrorists are presented here to illustrate the dilemmas facing the medical staff that treated them. It is suggested that international legal and bioethical guidelines are required to define the role of the physician and auxiliary medical staff vis a vis injured terrorists. There are extreme situations where the perpetration of violence and the defense of human rights come into conflict, leading to serious ethical and psychological discord. Terrorists, using violence to create fear in order to further their political objectives, might require life-saving medical care if injured during the course of their terror activities. PMID- 19998086 TI - Medical care of terrorists is "Beyond the letter of the law". PMID- 19998087 TI - The obligations of health workers to "terrorists". PMID- 19998088 TI - The terrorist and the doctor: a legal and ethical response. PMID- 19998089 TI - The psychology of repugnance and the duty to trust. PMID- 19998090 TI - No exceptionalism needed to treat terrorists. PMID- 19998091 TI - Ethical caring and the treatment of terrorists. PMID- 19998092 TI - Terrorists are just patients. PMID- 19998093 TI - Physician, where art thou? PMID- 19998094 TI - Separating the 'rights of' and 'justice for' bombers. PMID- 19998097 TI - Response to open peer commentaries on "Research 2.0: social networking and direct to-consumer personal genomics". PMID- 19998098 TI - A pragmatic consideration of ethical issues relating to personal genomics. PMID- 19998099 TI - Social networkers' attitudes toward direct-to-consumer personal genome testing. AB - PURPOSE: This study explores social networkers' interest in and attitudes toward personal genome testing (PGT), focusing on expectations related to the clinical integration of PGT results. METHODS: An online survey of 1,087 social networking users was conducted to assess 1) use and interest in PGT; 2) attitudes toward PGT companies and test results; and 3) expectations for the clinical integration of PGT. Descriptive statistics were calculated to summarize respondents' characteristics and responses. RESULTS: Six percent of respondents have used PGT, 64% would consider using PGT, and 30% would not use PGT. Of those who would consider using PGT, 74% report they would use it to gain knowledge about disease in their family. 34% of all respondents consider the information obtained from PGT to be a medical diagnosis. 78% of those who would consider PGT would ask their physician for help interpreting test results, and 61% of all respondents believe physicians have a professional obligation to help individuals interpret PGT results. CONCLUSION: Respondents express interest in using PGT services, primarily for purposes related to their medical care and expect physicians to help interpret PGT results. Physicians should therefore be prepared for patient demands for information and counsel on the basis of PGT results. PMID- 19998100 TI - It is settled: the 'is' can (and should) inform the 'ought'! PMID- 19998101 TI - Response to open peer commentaries on "What is the best standard for the standard of care in clinical research?". PMID- 19998102 TI - Personal genome testing: do you know what you are buying? PMID- 19998103 TI - Direct-to-consumer personal genome testing: the problem is not ignorance--it is market failure. PMID- 19998104 TI - Social networking and personal genomics: suggestions for optimizing the interaction. PMID- 19998105 TI - Genethics 2.0: phenotypes, genotypes, and the challenge of databases generated by personal genome testing. PMID- 19998106 TI - Are social networkers and genome testers one in the same? The limitations of public opinion research for guiding clinical practice. PMID- 19998107 TI - How attitudes research contributes to overoptimistic expectations of personal genome testing. PMID- 19998108 TI - Apomediation and the significance of online social networking. PMID- 19998109 TI - Personal genomics as an interactive web broadcast. PMID- 19998110 TI - We are the genes we've been waiting for: rational responses to the gathering storm of personal genomics. PMID- 19998111 TI - Challenges in the use of direct-to-consumer personal genome testing in children. PMID- 19998112 TI - Research 2.0: social networking and direct-to-consumer (DTC) genomics. AB - The convergence of increasingly efficient high throughput sequencing technology and ubiquitous Internet use by the public has fueled the proliferation of companies that provide personal genetic information (PGI) direct-to-consumers. Companies such as 23andme (Mountain View, CA) and Navigenics (Foster City, CA) are emblematic of a growing market for PGI that some argue represents a paradigm shift in how the public values this information and incorporates it into how they behave and plan for their futures. This new class of social networking business ventures that market the science of the personal genome illustrates the new trend in collaborative science. In addition to fostering a consumer empowerment movement, it promotes the trend of democratizing information--openly sharing of data with all interested parties, not just the biomedical researcher--for the purposes of pooling data (increasing statistical power) and escalating the innovation process. This target article discusses the need for new approaches to studying DTC genomics using social network analysis to identify the impact of obtaining, sharing, and using PGI. As a locus of biosociality, DTC personal genomics forges social relationships based on beliefs of common genetic susceptibility that links risk, disease, and group identity. Ethical issues related to the reframing of DTC personal genomic consumers as advocates and research subjects and the creation of new social formations around health research may be identified through social network analysis. PMID- 19998113 TI - Direct-to-consumer genomics, social networking, and confidentiality. PMID- 19998114 TI - Personal genomics: democratization, or empowerment, or 'something'. PMID- 19998115 TI - Direct-to-consumer genetics and health policy: a worst-case scenario? PMID- 19998116 TI - Networking genetics, populations, and race. PMID- 19998117 TI - Genetic privacy: might there be a moral duty to share one's genetic information? PMID- 19998118 TI - Direct-to-consumer genome-wide scans: astrologicogenomics or simple scams? PMID- 19998119 TI - Direct-to-consumer genomics and research ethics: should a more robust informed consent process be included? PMID- 19998121 TI - Categorizing empirical research in bioethics: why count the ways? PMID- 19998120 TI - The role of empirical research in bioethics. AB - There has long been tension between bioethicists whose work focuses on classical philosophical inquiry and those who perform empirical studies on bioethical issues. While many have argued that empirical research merely illuminates current practices and cannot inform normative ethics, others assert that research-based work has significant implications for refining our ethical norms. In this essay, I present a novel construct for classifying empirical research in bioethics into four hierarchical categories: Lay of the Land, Ideal Versus Reality, Improving Care, and Changing Ethical Norms. Through explaining these four categories and providing examples of publications in each stratum, I define how empirical research informs normative ethics. I conclude by demonstrating how philosophical inquiry and empirical research can work cooperatively to further normative ethics. PMID- 19998122 TI - Answering the 'so what?' question for empirical research in bioethics. PMID- 19998123 TI - Kon's reinvention of the empirical bioethics wheel. PMID- 19998124 TI - What counts as empirical research in bioethics and where do we find the stuff? PMID- 19998125 TI - What empirical research can do for bioethics. PMID- 19998126 TI - Empirical bioethics: present and future possibilities. PMID- 19998127 TI - Beware dichotomies and grand abstractions: attending to particularity and practice in empirical bioethics. PMID- 19998128 TI - Conceptualizing disease: unifying the divide between philosophical inquiry and empirical research. PMID- 19998129 TI - Psychology and medical decision-making. PMID- 19998130 TI - An engine, not a pyramid. PMID- 19998131 TI - How to build a theory about empirical bioethics: acknowledging the limitations of empirical research. PMID- 19998132 TI - Evidence-based research ethics. PMID- 19998133 TI - A view from the borderlands of philosophical bioethics and empirical social science research: how the 'is' can inform the 'ought'. PMID- 19998134 TI - Empiricism, values, and bioethics. PMID- 19998135 TI - Methodology, epistemology, and empirical bioethics research: a constructive/ist commentary. PMID- 19998136 TI - Hurrah for empirical bioethics (where hermeneutically clarified) or how perception of facts 'depends' on values. PMID- 19998137 TI - Improving care and ethics: a plea for interactive empirical ethics. PMID- 19998138 TI - Empirical bioethics research in the developing world: when the 'is' is close to an 'ought'. PMID- 19998141 TI - Return of "accurate" and "actionable" results: yes! PMID- 19998142 TI - Response to open peer commentaries on "Why consent may not be needed for organ procurement". PMID- 19998143 TI - Sir John Maddox and the ethics of heresy. PMID- 19998144 TI - Disclosing clinical trial results: publicity, significance and independence. PMID- 19998145 TI - Why consent may not be needed for organ procurement. AB - Most people think it is wrong to take organs from the dead if the potential donors had previously expressed a wish not to donate. Yet people respond differently to a thought experiment that seems analogous in terms of moral relevance to taking organs without consent. We argue that our reaction to the thought experiment is most representative of our deepest moral convictions. We realize not everyone will be convinced by the conclusions we draw from our thought experiment. Therefore, we point out that the state ignores consent in performing mandatory autopsies in some cases. If readers are willing to give up the permissibility of mandatory autopsies, we then offer some metaphysical arguments against posthumous harm. Drawing upon claims about bodies ceasing to exist at death and Epicurean-inspired arguments against posthumous interests, we make a case for an organ conscription policy which respects fundamental liberal principles of autonomy, bodily integrity, and property. PMID- 19998146 TI - Response to open peer commentaries on "Prenatal diagnosis and abortion for congenital abnormalities: is it ethical to provide one without the other?". PMID- 19998147 TI - Pardon my asking: what's new? PMID- 19998148 TI - The organ conscription trolley problem. PMID- 19998149 TI - Hypotheticals, analogies, death's harms, and organ procurement. PMID- 19998150 TI - Survivors' interests in human remains. PMID- 19998151 TI - Why intuitions and metaphysics are the wrong approach for health law: a commentary on Delaney and Hershenov. PMID- 19998152 TI - Is context a distortional factor, really? PMID- 19998153 TI - Advancing the case for organ procurement. PMID- 19998154 TI - The duty to disclose adverse clinical trial results. AB - Participants in some clinical trials are at risk of being harmed and sometimes are seriously harmed as a result of not being provided with available, relevant risk information. We argue that this situation is unacceptable and that there is a moral duty to disclose all adverse clinical trial results to participants in clinical trials. This duty is grounded in the human right not to be placed at risk of harm without informed consent. We consider objections to disclosure grounded in considerations of commercial interest, and we argue that these concerns are insufficient to override the moral duty to disclose adverse clinical trial results. However, we also develop a proposal that enables commercial interests to be protected, while promoting the duty to disclose adverse clinical trial results. PMID- 19998155 TI - The duty to disclose (even more) adverse clinical trial results. PMID- 19998156 TI - Compounding risks to patients: selective disclosure is not an option. PMID- 19998157 TI - Improving subject recruitment by maintaining truly informed consent: a practical benefit of disclosing adverse clinical trial results. PMID- 19998158 TI - Respecting participant autonomy and the disclosure of clinical trial results. PMID- 19998159 TI - Enhancing informed consent in clinical trials and exploring resistances to disclosing adverse clinical trial results. PMID- 19998160 TI - The duty to register phase I trials. PMID- 19998161 TI - Precautionary harm disclosure in clinical trials. PMID- 19998162 TI - Disclosure of adverse clinical trial results--should legal immunity be granted to drug companies? PMID- 19998163 TI - Prenatal diagnosis and abortion for congenital abnormalities: is it ethical to provide one without the other? AB - This target article considers the ethical implications of providing prenatal diagnosis (PND) and antenatal screening services to detect fetal abnormalities in jurisdictions that prohibit abortion for these conditions. This unusual health policy context is common in the Latin American region. Congenital conditions are often untreated or under-treated in developing countries due to limited health resources, leading many women/couples to prefer termination of affected pregnancies. Three potential harms derive from the provision of PND in the absence of legal and safe abortion for these conditions: psychological distress, unjust distribution of burdens between socio-economic classes, and financial burdens for families and society. We present Iran as a comparative case study where recognition of these ethical issues has led to the liberalization of abortion laws for fetuses with thalassemia. We argue that physicians, geneticists and policymakers have an ethical and professional duty of care to advocate for change in order to ameliorate these harms. PMID- 19998164 TI - A closer look at the abortion debate in Iran. PMID- 19998165 TI - Prenatal diagnosis and abortion are not in conflict in Israel. PMID- 19998166 TI - From Iran to Latin America: must prenatal diagnosis necessarily be provided with abortion for congenital abnormalities? PMID- 19998167 TI - An unjustified exception to an unjust law? PMID- 19998168 TI - Does prenatal diagnosis morally require provision of selective abortion? PMID- 19998169 TI - Can, should, ought, must. PMID- 19998170 TI - Prenatal testing can be advantageous even when abortion is not an option. PMID- 19998171 TI - Abortion: prohibitions and exceptions. PMID- 19998175 TI - Responses to open peer commentaries on "The Subjective Brain, Identity, and Neuroethics". PMID- 19998176 TI - Neurologisms. PMID- 19998177 TI - Mirror neurons and the reenchantment of bioethics. PMID- 19998178 TI - Reviewing the unsubstantiated claims for the methodological superiority of 'placebo' over 'active' controlled trials: reply to open peer commentaries. PMID- 19998179 TI - The subjective brain, identity, and neuroethics. AB - The human brain is subjective and reflects the life of a being-in-the-world-with others whose identity reflects that complex engaged reality. Human subjectivity is shaped and in-formed (formed by inner processes) that are adapted to the human life-world and embody meaning and the relatedness of a human being. Questions of identity relate to this complex and dynamic reality to reflect the fact that biology, human ecology, culture, and one's historic-political situation are inscribed in one's neural network and have configured its architecture so that it is a unique and irreplaceable phenomenon. So much is a human individual a relational being whose own understanding and ownership of his or her life is both situated and distinctive that neurophilosophical conceptions of identity and human activity that neglect these features of our being are quite inadequate to ground a robust neuroethics. PMID- 19998180 TI - Response to Muller. Body integrity identity disorder (BIID). PMID- 19998181 TI - Subjectivity, consciousness, and pain: the importance of thinking phenomenologically. PMID- 19998182 TI - A descriptive social neuroethics is needed to reveal lived identities. PMID- 19998183 TI - Stigma and addiction: being and becoming. PMID- 19998184 TI - Skills, dementia, and bridging divides in neuroscience. PMID- 19998185 TI - Transferring emerging neuroscience to the clinical ethics bedside. PMID- 19998186 TI - Subjectivity, the brain, life narratives and the ethical treatment of persons with Alzheimer's disease. PMID- 19998187 TI - Brain, body, and society: bioethical reflections on socio-historical neuroscience and neuro-corporeal social science. PMID- 19998188 TI - What are the subjective processes in our brain? Empirical and ethical implications of a relational concept of the brain. PMID- 19998189 TI - From the subjective brain to the situated person. PMID- 19998190 TI - The subjective brain, identity, and neuroethics: a legal perspective. PMID- 19998191 TI - The place of moral responsibility and mental illness. PMID- 19998192 TI - Questioning the methodologic superiority of 'placebo' over 'active' controlled trials. AB - A resilient issue in research ethics is whether and when a placebo-controlled trial (PCT) is justified if it deprives research subjects of a recognized treatment. The clinicians' moral duty to provide the best available care seems to require the use of 'active' controlled trials (ACTs) that use an established treatment as a control whenever such a therapy is available. In another regard, ACTs are supposedly methodologically inferior to PCTs. Hence, the moral duty of the clinical researcher to use the best methods will favor PCTs. In this target article, I analyze the three reasons for believing that ACTs are inferior to PCTs namely: 1) ACTs lack 'assay sensitivity'; 2) ACTs do not measure absolute effect size; and 3) ACTs require more participants; and I contend that none are acceptable. Consequently the tension between clinical and research ethics dissolves: the moral duty of the clinician to avoid PCTs is unopposed by methodological considerations. PMID- 19998193 TI - The rationale for placebo-controlled trials: methodology and policy considerations. PMID- 19998194 TI - Preparing for a post-placebo paradigm: ethics and choice of control in clinical trials. PMID- 19998195 TI - Placebo misconceptions. PMID- 19998196 TI - A clinical perspective on placebo research: looking back, looking forward. PMID- 19998197 TI - Ethical issues in the difference between placebo-controlled and active-controlled trials. PMID- 19998198 TI - Conflating scientific with clinical considerations. PMID- 19998199 TI - Who's in control of the choice of control? PMID- 19998200 TI - Strengthening Howick's argument against the alleged superiority of placebo controlled trials. PMID- 19998201 TI - Research, medicine, and "placebos". PMID- 19998202 TI - Questioning the methodological superiority of 'placebo' over 'active' controlled trials. PMID- 19998203 TI - Control theory: placebo-controlled drug trials have problems. Active-controlled drug trials are not always the solution. PMID- 19998205 TI - High-frequency 3D visualization of an embryo early in the first trimester (8 + 2 weeks of gestation). PMID- 19998206 TI - Quality assurance in ultrasound diagnostics--where are we at the end of 2009? PMID- 19998207 TI - Ultrasound imaging of abdominal aortic aneurysms: diagnosis of aneurysms and complications and follow-up after endovascular repair. PMID- 19998208 TI - Correlation between SonoVue enhancement in CEUS, HCC differentiation and HCC diameter: analysis of 130 patients with hepatocellular carcinoma (HCC). AB - PURPOSE: The aim of the study was to characterize SonoVue enhancement in hepatocellular carcinoma in correlation to both lesion diameter and histological differentiation of the lesion. MATERIALS AND METHODS: In a prospective study 130 patients (72 male, 58 female, 62 +/- 10 years) with HCC lesions detected by B mode sonography were examined. After injection of 1.2 - 2.4 ml SonoVue, HCC lesions were examined continuously for up to 5 min using "low MI" sonography. RESULTS: Arterial hypervascularization was found in 72 % of the HCC lesions without correlation to the lesion diameter or histological grading, when analyzed for the total group. However, the analysis of the G 1 subgroup showed significant correlation between lesion diameter and arterial hypervascularization. Arterial hypervascularization was found in 95 % of the G 1 lesions > 3 cm but in only 43 % of the G 1 lesions < 3 cm (p < 0.001). In contrast, analysis of the remaining HCC lesions (without G 1) showed arterial hypervascularization in 69 % of the lesions < 3 cm and in 72 % of the lesions > 3 cm (n. s.) without correlation to the diameter. In the late phase in the G 1 subgroup, hypoechoic demarcation was found in 95 % of the G 1 lesions > 3 cm, but in only 64 % of the G 1 lesions < 3 cm (p < 0.001). In contrast, in the less differentiated HCC lesions (without G 1), hypoechoic demarcation was found in 91 % (HCC > 3 cm) and in 82 % (HCC < 3 cm) of the lesions (n. s.). CONCLUSION: In well-differentiated HCCs (G1) hyperechoic enhancement in the arterial phase and hypoechoic demarcation in the late phase correlate to the diameter. PMID- 19998209 TI - Diagnostic value of palpation and ultrasonography for diagnosing breast cancer recurrence after mastectomy--a comparison. AB - PURPOSE: According to the current interdisciplinary S 3 guideline for breast cancer diagnostics, treatment, and follow-up (1st update 2008), palpation is one of the diagnostic mainstays for follow-up examinations after mastectomy. Although recommended in the manuscript, regular ultrasonographic examinations are neither explicitly mentioned in the statement nor in the follow-up plan. In ambiguous cases, MRI can serve as a supplementary diagnostic method. In order to evaluate the value of palpation and ultrasonography for diagnosing recurrent disease after mastectomy, we analyzed the sensitivity of each method individually and the sensitivity of both methods combined. MATERIALS AND METHODS: Over a 12-year time span, histological data from 57 patients suspected to have recurrent disease after mastectomy (benign lesions: n = 15, 26.3% malignant lesions: n = 42, 73.7%) were collected. Prior to biopsy, the benign versus malignant character of all lesions was assessed by palpation and ultrasonography (BI-RADS). Sensitivity, specificity, positive predictive value (ppv), negative predictive value (npv), and efficacy were calculated using a contingency table. RESULTS: Palpation had a sensitivity of 85.7 % and a specificity of 6.7% the respective figures for ultrasonography were 90.5% and 46.7%. The sensitivity of palpation and ultrasonography combined was 100%, i. e. 14.3% higher than the sensitivity of palpation alone. Five cases of recurrent disease which remained undetected by palpation were only recognized by ultrasonography. CONCLUSION: In agreement with several other studies, the results of our retrospective study show that ultrasonography is superior to palpation for diagnosing recurrent disease after mastectomy. Prospective multicenter studies are needed to evaluate ultrasonographic follow-up before an amendment of the current S 3 guidelines can be recommended with a high degree of evidence. PMID- 19998210 TI - Pentalogy of Cantrell vs. limb body wall complex: differential diagnosis of a severe malformation in early pregnancy. PMID- 19998211 TI - [Olfactory dysfunction: correlation of olfactory bulb volume on MRI and objective olfactometry]. AB - PURPOSE: To define the role of olfactory bulb volume measurement by magnetic resonance imaging (MRI) for detecting olfactory dysfunction in comparison with objective olfactometry. MATERIALS AND METHODS: Thirty patients with suspected olfactory dysfunction (16 women, 14 men; mean age 52 years, range 20 - 79 years) were examined by MRI and objective olfactometry between January 2006 and January 2009. Olfactory bulb volumes were measured by two neuroradiologists using 3D MR data sets. The olfactory function was categorized as normosmia, hyposmia, and anosmia on the basis of objective olfactometry. Pearson correlation coefficients were calculated for objective olfactometry and olfactory bulb volumes on MRI. ROC analysis was performed to determine whether MRI bulb volumes can serve to predict anosmia or hyposmia. RESULTS: The bulb volumes measured by MRI ranged from 0 to 135.9 mm (3). Based on olfactometry, anosmia was present in 11 patients (total bulb volume of 15.7 +/- 23.3 mm (3)), hyposmia in 9 patients (total bulb volume of 50.0 +/- 25.5 mm (3)), and normosmia in 10 patients (total bulb volume of 110.7 +/- 21.5 mm (3)). There was good correlation (r > 0.9) between objective olfactometry and olfactory bulb volume on MRI. ROC analysis yielded a cut-off value of 32 mm (3) for anosmia, which had a sensitivity of 0.91 and specificity of 0.947. The cut-off value for olfactory dysfunction was 80.7 mm (3) (sensitivity 0.95; specificity of 0.9). CONCLUSION: The olfactory bulb volume determined by MRI is a suitable parameter for diagnosing complete or partial loss of the sense of smell. PMID- 19998212 TI - [Rarely bilateral: hypertrophic degeneration of the olivary nucleus]. PMID- 19998213 TI - [Temporary transvenous embolization of a giant arteriovenous fistula of the scalp -a preoperative option]. PMID- 19998214 TI - [Detection of a neuroendocrine differentiated cystic pancreatic lesion by gallium 68-DOTATOC-PET/CT with inconclusive MRI, CT and ultrasound diagnosis]. PMID- 19998216 TI - [Phenotyping specific language impairment in kindergarten children]. AB - OBJECTIVE: For phenotyping specific language impairment (SLI) in kindergarten children in clinical practice and research issues, we need a valid diagnostic method for dichotomous classification (language impaired, normal developing). PATIENTS AND METHODS: 27 kindergarten children belonged to SLI-group, 36 to control-group. The diagnostic accuracy of a composed language test battery was examined in comparison to a clinical assessment. The test battery was composed of 8 subtests of German norm-referenced, standardized tests. RESULTS: Several discriminant analyses showed acceptable levels of accuracy with over 80% for sensitivity and specificity. Using a single subtest the subtest "Phonologisches Arbeitsgedachtnis fur Nichtworter" (phonological short-term memory of nonwords) from Sprachentwicklungstest fur drei- bis funfjahrige Kinder (SETK 3-5; Grimm, 2001) showed best classification rates between the two groups using a Cut-off point of -0,39 SD. Means of the 8 used subtests showed significant differences for the two groups. CONCLUSION: The described method for phenotyping SLI can identify children with normal language and those with impaired language with acceptable levels of diagnostic accuracy. When using a norm-referenced standardized test for the assessment of language abilities, it is important to have empirically derived information about diagnostic accuracy (sensitivity, specificity, Cut-off score). PMID- 19998217 TI - [Morphologic classification of Reinke's edema through optical coherence tomography]. AB - OBJECTIVE: Several classification systems for Reinke's edema have been proposed in the past, which are somewhat less than morphologically ideal. The objective of the present study is to evaluate the ability of optical coherence tomography (OCT) to attain a reproducible graduation of Reinke's edema. MATERIAL AND METHODS: In a prospective study, 30 consecutive patients underwent endolaryngeal, microsurgical resection of their Reinke's edema. Graduation was performed through OCT based on morphologic criteria, where each result was compared with that of other classification systems. RESULTS: In Reinke's edema grade I according to Glanz, a feathered pattern is found, while Reinke's edema grade II demonstrates a lacunar and Reinke's edema grade III a confluent pattern. Correlation between the different classification systems was uniformly weak and not significant. CONCLUSIONS: Due to the three different recognizable patterns, OCT qualifies as an objective and reproducible aid for the graduation of Reinke's edema and gives certain insights in its pathogenesis. PMID- 19998218 TI - Safety and efficacy of cytology brushings versus standard fine-needle aspiration in evaluating cystic pancreatic lesions: a controlled study. AB - BACKGROUND AND STUDY AIMS: Cystic pancreatic lesions (CPLs) are increasingly detected by various imaging studies. Mucinous CPLs carry a risk of malignant transformation but this is often difficult to diagnose preoperatively. In a previous report of 10 suspected mucinous CPLs, the cellular yield of endoscopic ultrasonography (EUS)-guided cytology brushings was found to be superior to the yield from standard fine-needle aspiration (FNA). The aim of this prospective and blinded study was to compare the cytology yield of mucinous epithelium from brushing with FNA in suspected mucinous CPLs. PATIENTS AND METHODS: In total, 37 patients with 39 CPLs measuring at least 20 mm were enrolled between June 2006 and July 2008 for EUS-cytobrushing and EUS-FNA of CPLs. Demographic, clinical, EUS, cytopathologic, and surgical data were recorded whenever available. Yield of cytology brushings was compared with that of FNA. Procedure morbidity was evaluated after 30 days. The main outcome assessed was yield of intracellular mucin (ICM) on cytobrushing specimens compared with EUS-FNA for the diagnosis of suspected mucinous CPL. RESULTS: Cytobrushings were more likely to detect ICM than the EUS-FNA method ( P = 0.001). In three patients with hypocellular FNA, dysplasia was found on cytology brushing and later confirmed by surgical pathology. Significant complications occurred in three patients (8 %): one postbrushing bleeding and two acute pancreatitis. CONCLUSIONS: Cytology brushings are more likely to provide an adequate mucinous epithelium specimen than standard FNA and could aid the diagnosis of CPLs in a selective group of patients. PMID- 19998219 TI - Endoscopic treatment of difficult bile duct stones by using a double-lumen basket for laser lithotripsy--a case series. AB - Laser lithotripsy is an effective and safe method for difficult common bile duct (CBD) stones. However, radiolucent laser fibers make accurate fragmentation difficult and require continuous visual control or an accessory for effective targeting. The newly developed double-lumen basket may promote effective laser lithotripsy after stone capture. We performed laser lithotripsy using a double lumen basket in 14 patients with CBD stones refractive to conventional endoscopic treatment, and evaluated the feasibility and efficacy of this procedure. Stones were successfully fragmented in 13 of 14 patients, and 13 patients eventually became stone-free. Mechanical lithotripsy was applied in two patients with biliary strictures. Minor complications were noted in three patients, including transient hemobilia in one patient. For a selected group of patients with difficult CBD stones, laser lithotripsy using a double-lumen basket appears to be an effective and safe method. However, continuous development of basket protocols to increase the success rate of lithotripsy is needed. PMID- 19998220 TI - [Mucocele of the appendix - a heterogenous surgical pathology]. AB - BACKGROUND: Mucoceles of the appendix are rare. After appendectomy, mucoceles are detected with a frequency of 0.2 to 0.3 %. Both stenosing / obliterating processes and alterations of the epithelium (hyperplasia, mucinous cystadenoma, cystadenoma with uncertain malignant potential (UMP), mucinous cystadenocarcinoma lead to the occurrence of mucoceles. The perforation of a mucocele with possible spread of mucus and cells into the abdominal cavity constitutes a severe complication (pseudomyxoma peritonei). Surgical resection is the curative approach for mucoceles of the appendix. MATERIALS AND METHODS: Data of patients who were treated for an appendiceal mucocele between 1995 and 2009 were analysed retrospectively with regard to clinical presentation, diagnostic measures, surgical procedure and histopathological result. Follow-up was evaluated in telephone interviews. RESULTS: We extracted 5 cases from our database. Clinical symptoms varied greatly among the individual patients, ranging from peracute abdominal pain in the right lower quadrant to chronic obstipation. Results from abdominal ultrasound and / or abdominal CT scans contributed to the indication for surgical intervention in all cases. In 2 patients surgery was stated as urgent whereas in 3 the operation was scheduled electively. In one patient the diagnosis of an appendiceal mucocele was stated preoperatively and in another intraoperatively. In 3 patients only the histopathological result revealed the underlying mucocele. We performed 1 open and 1 laparoscopic appendectomy, 1 open appendectomy with a partial resection of the coecum and 2 laparoscopic ileocoecal resections. One of the patients had a pseudomyxoma peritonei. The histopathological diagnoses ranged from mere epithelial hyperplasia to an adenoma with uncertain malignant potential and a mucinous cystadenocarcinoma. One patient's long-term follow-up could not be evaluated. All other patients had neither recurrence nor any complications after discharge. CONCLUSIONS: Mucoceles of the appendix present with a wide spectrum of clinical symptoms and histopathological alterations. Only an accurate histological analysis reveals the underlying pathological lesion correctly. This study emphasises that a mucocele of the appendix constitutes an important differential diagnosis in patients presenting with pathologies in their right lower abdominal quadrant. PMID- 19998221 TI - [Evolution in the operating room]. PMID- 19998222 TI - [Bipolar hemiarthroplasty in femoral neck fractures--impact of duration of surgery, time of day and the surgeon's experience on the complication rate]. AB - AIM: Bipolar hemiarthroplasty is frequently used in femoral neck fractures. There is only little evidence regarding differences in complication rates if the procedure is performed by either a junior or senior surgeon. METHODS: 360 bipolar hemiarthroplasties were retrospectively investigated. Complication rates for junior and senior surgeons were evaluated as well as differences between daytime and nightshift surgery. We also assessed the duration of surgery for the two groups and its impact on the complication rate. RESULTS: The average duration of the procedure was 67 minutes (23-194) with statistically significant differences between junior and senior surgeons (77 vs. 61 minutes, p < 0.001). Complications occurred in 27 (7.5%) of all cases. Postoperative infections were most frequently found (3.1% of all complications), followed by haematomas and dislocations of the implant (1.7% and 1.1 %, respectively). 25 patients required revision operations. More complications were found in cases performed by junior surgeons (9.56% vs. 6.25%). This difference was not significant (p = 0.248). During on-call duty we also observed more complications compared to daytime surgery (11% vs. 7%). There was no coherence between the duration of surgery and the incidence of complications. CONCLUSIONS: Bipolar hemiarthroplasty is a reliable treatment option for femoral neck fractures even when performed by a junior surgeon. The higher incidence of complications during nighttime surgery should be a reason to perform those cases that are not urgent during the daytime shift. PMID- 19998223 TI - [Cage assisted fusion operation of the spine]. PMID- 19998224 TI - [Management of adolescents with childhood onset growth hormone deficiency in the transition--results of a field based study in Germany]. AB - BACKGROUND: Discontinuation of growth hormone (GH) treatment upon attainment of final height has been associated with impaired somatic development and altered body composition. Therefore, optimal care of patients with GH deficiency (GHD) in the transition phase from adolescence to adulthood is a challenge for all parties involved. We analyzed the current clinical practice in Germany. METHODS: In 2008, 124 endocrinologists (69 pediatric, 55 adult endocrinologists) in Germany were interviewed using a structured questionnaire. RESULTS: Overall, 67 % of pediatric endocrinologists (PE) and adult endocrinologists (AE) declared to have a contact physician for their patients. 13 endocrinologists declared to have a common transition clinic with their corresponding colleague. 74 % of PE stated to transfer their patients after the end of GH therapy to an AE. 62 % of the patients were transferred at the age of 18 years. 70 % of the PE stated to retest their patients themselves, while 70 % of the AE answered that the patients had not been retested when they first came to the adult clinic. For the evaluation of GH-secretion, PE most frequently used the arginine (86 %), ITT (35 %) and clonidine test (33 %), whereas AE utilized the GHRH/arginine test (71 %), and the ITT (67 %). The level of patient's information about his disease status was considered as "good" by 44 % of AE (77 % by AE having established a transition clinic). The quality of patient files transferred from the PE was considered as "good" by 54 % of all AE (100 % by AE with transition clinic). CONCLUSION: To a significant extent, there is an inconsistence in diagnostic methods and treatment modalities performed by PE and AE compared to recently published consensus guidelines. Only 13 PE interviewed in this study transfer their GHD patients in a transition clinic setting. Communication and transfer of information between both groups appears to be impaired in centres without a transition clinic. In those clinics having established transition clinics, patient's status of information and quality of patient files is considered to be much better. PMID- 19998225 TI - [Lead intoxication in a group of workers in Germany]. AB - HISTORY AND ADMISSION FINDINGS: Seventeen East-European workers with a suspected lead-intoxication presented themselves to the Department of Toxicology. All of them had worked on the renovation of pylons of a high-tension line. The old paint, known to contain lead was removed with needle descalers. The patients had blood lead concentrations between 325 and 1124 microg/l, but no specific symptoms. The workers neglected the protective measures at their working-place. INVESTIGATIONS: 12 of 17 workers had lead-concentrations above 400 microg/l (Reference < 90 microg/l). 10 of 17 patients showed an increased level of free protoporphyrins and all workers showed a decreased activity of delta aminolaevulinacid-dehydratase (ALAD). TREATMENT AND COURSE: Patients with lead concentration above 700 microg/l were treated with the chelating agent meso-2,3 dimercaptosuccinic acid (DMSA) 3 x 200 mg/d for nine days. The patients with lead concentrations between 400 and 700 microg/l were treated which DMSA 3 x 100 mg/d. After the DMSA-treatment the lead-concentrations had dropped (p < 0.001). During the DMSA-therapy one patient had to be treated in the hospital because of a generalised allergic exanthema. CONCLUSION: We report seventeen patients with high lead concentration in their blood due to occupational exposure. The high blood lead levels showed that the workers had not been protected adequately. This examplifies that occupational lead exposure still occurs, also in Germany. By patients with unspecific symptoms connected with lead exposure a biomonitoring for lead is necessary. PMID- 19998226 TI - [70-year-old patient with conspicuities at the forearm]. PMID- 19998227 TI - [Diagnosis and treatment in hereditary hemochromatosis]. PMID- 19998228 TI - [Acute lung injury: principles and perspectives]. PMID- 19998229 TI - [Pre-exposure prophylaxis in HIV--a vision or soon a reality?]. AB - Condom use is propagated as the most efficient measure to prevent HIV transmission. For several reasons, condoms are NOT ALWAYS used or misapplied during sexual intercourse. Therefore, alternative preventive measures through intake of antiretroviral drugs before sexual intercourse with a (presumably) HIV positive person are being considered, so called Pre-Exposure Prophylaxis (PrEP). In animal models the efficacy of HIV-PrEP was shown for Tenofovir alone or in combination with Emtricitabine). Several clinical studies are currently being conducted in different HIV-risk groups on various continents. First results from these studies are anticipated for the year 2010. In case of proven efficacy for HIV-PrEP, our health system would face a large interdisciplinary challenge. It would be a difficult task to define the appropriate recipients. Measures would have to be taken to limit possible misuse of antiretroviral drugs, due to the negative consequences with development of resistance, adverse events and illegal trading. It is already evident that HIV-PrEP will not provide absolute protection, nor will it replace other preventive strategies. However, if used cautiously, HIV-PrEP might be established as a useful supplement in the prevention of HIV. Paramount questions from the fields of epidemiology, behavioural science, logistics, health politics and ethics should be answered in advance. PMID- 19998230 TI - [Ischemia and reperfusion: options for pharmacological modulation with natriumselenit]. PMID- 19998231 TI - [Can the results of plastic surgery be improved by the modulation of reperfusion damage?]. PMID- 19998232 TI - [Biochemistry and pathophysiology of ischemia and reperfusion]. PMID- 19998233 TI - [Clinical data on the use of selenase in sepsis]. PMID- 19998234 TI - [Ischemia and reperfusion in vascular surgery interventions--visible and invisible risks]. PMID- 19998235 TI - [Selenium substitution after cardiac arrest]. PMID- 19998236 TI - [Reperfusion and potential damages in heart surgery]. PMID- 19998237 TI - [Transplantation: potential indication for antioxidants?]. PMID- 19998238 TI - Transient salt wasting in POMC-deficiency due to infection induced stress. AB - Obesity is a multifactorial disorder influenced by genetic, behavioral, environmental and cultural factors. A twelve month old male patient was admitted to the hospital because of malaise, irritability, disquietness and obesity. His BMI was 19.8 kg/m (2) and BMI SDS was 1.38. Mental development was normal, and motor skills were mildly delayed most probably due to his obesity. His physical examination was totally normal except obesity and red hair. A history of hypoglycemia on the fourth day of life, which resolved after oral glucose administration, was reported. The child had been hyperphagic from the first weeks of life and had aggressive behavior when food was denied. The body weight of the patient increased dramatically during the first year of life. Based on the clinical features and laboratory findings (the overgrowth syndrome, red hair, hypoglycemia and hypocortisolism) the patient was diagnosed as POMC deficiency and the diagnosis was confirmed by genetic studies. Hypoglycemia and apnea episodes ceased as he was put on hydrocortisone but he developed relative mineralocorticoid deficiency during a urinary tract infection. In POMC deficiency, relative mineralocorticoid deficiency should be in mind in episodes of severe stress and therapy should be initiated. PMID- 19998239 TI - False positive test results for pheochromocytoma from 2000 to 2008. AB - Testing for pheochromocytoma becomes more frequent in clinical practice. False positive test results may cause patient anxiety and unnecessary imaging studies. The data on false positive results for pheochromocytoma in routine clinical practice are lacking. To examine the prevalence of false positive results and to reveal the clinical features and laboratory tests of patients with markedly elevated but false positive test results, a database of tests for pheochromocytoma at a large general hospital between 2000 and 2008 was reviewed. Of 1,896 patients tested, 417 (22.0%) had at least one abnormal test result and 66 (3.5%) had markedly elevated results. 24 patients with markedly elevated but false positive results received 65 imaging studies and 1 adrenalectomy. The causes of the misleading results included physiological variations (33%), laboratory errors (29%), and drug interference with measurement (21%). The false positive rate of a markedly elevated result was lowest for vanillylmandelic acid (9%) and highest for urine metanephrines (50%) (p = 0.03). Nearly half of all test results were normal and 79% of patients had at least one normal result. Therefore false positive test results for pheochromocytoma are rather common. Markedly elevated but false positive test results can potentially be avoided by judicious selection of patients and tests. Pretest risk, physiological variations of catecholamine levels, laboratory errors, and drug interference should be considered in interpreting abnormal test results. PMID- 19998240 TI - Effects of glycyrrhizic acid on 11 beta-hydroxysteroid dehydrogenase (11 betaHSD1 and 2) activities and HOMA-IR in rats at different treatment periods. AB - Glycyrrhizic acid (GA) has been reported to inhibit postprandial blood glucose rise and 11 beta-hydroxysteroid dehydrogenase 1 (11 betaHSD1) activity. As not much work has been done on GA effects on 11 betaHSD1 and 2 and HOMA-IR at different treatment periods, this work was conducted. 60 male Sprague Dawley rats fed AD LIBITUM were assigned into six groups of control and treated that were given GA at different duration namely 12, 24 and 48 h. Treated and control groups were intraperitoneally administered with GA (50 mgkg (-1)) and saline respectively. Blood and subcutaneous (ATS) and visceral adipose tissue (ATV), abdominal (MA) and quadriceps femoris muscle (MT), liver (L) and kidney (K) were examined. HOMA-IR in GA-treated rats decreased in all groups (P<0.05). In the 12 h and 24-h treated rats, 11 betaHSD1 activities decreased in all tissues (P<0.05) except MA and MT (P>0.05) in the former and ATV (P>0.05) in the latter. However, 11 betaHSD1 activities decreased significantly in all tissues ( P<0.05) in the 48 h treated rats. Significant decrease in 11 betaHSD2 (P>0.05) activities were observed in the L of all treatment groups and K in the 24-h and 48-h treated rats (P<0.05). Histological analysis on ATS showed increase in the number of small size adipocytes while ATV adipocytes showed shrinkage after GA administration. Increased glycogen deposition in the L was observed in the GA-administered rats in all the treatment periods. In conclusion, GA treatment showed a decrease in the HOMA-IR and both 11 betaHSD1 and 2 activities in all tissues, with more profound decrease in the 48-h treated rats. PMID- 19998241 TI - Does normal thyroid gland by ultrasonography match with normal serum thyroid hormones and negative thyroid antibodies? AB - Few papers have shown that a hypoechoic appearance of the thyroid gland at ultrasonography (US) is related to a hypofunction and serum positivity of thyroid antibodies (T-Ab). However, it is not ascertained if normal thyroid appearance at US correspond to normal thyroid laboratory tests. The aim of this study was to assess the value of normal thyroid at US in predicting normal thyroid hormones and negative T-Ab in a cohort of 48 adult patients. All patients (37 females and 11 males) were referred to our hospital to undergo their first thyroid US examination, followed by a thyroid function evaluation. All subjects had normal thyroid gland at US. As a control group 65 patients with hypoechoic and inhomogeneous thyroid gland were enrolled. All 48 patients had normal free-T (3) and free-T (4) levels. While 41 patients (85.4%) showed normal TSH, in 7 subjects (14.6%) TSH was elevated and a significant (p < 0.001) difference was recorded between the two groups in mean TSH value. Positive T-Ab value was found in 5 patients (10.4%) and the remaining 43 patients (89.6%) had negative T-Ab. TSH was not significantly correlated with age, thyroid volume or BMI. The multivariate model showed that only BMI was significantly correlated to thyroid volume (p < 0.01, r(2)=0.31). These results showed that normal thyroid recorded by US matches with normal thyroid laboratory assessment to a large degree. These preliminary data need to be confirmed in a prospective study and in a larger series and should suggest the evaluation of thyrotropin and thyroid antibodies in subjects with normal thyroid gland as assessed by US. PMID- 19998242 TI - Effect of glucocorticoid-, parathyroid- and thyroid hormones excess on human iliac crest bone matrix insulin-like growth factor (IGF)-I in patients with osteoporosis. AB - Insulin-like growth factor-I (IGF-I) is a well documented bone-active growth factor. Clinical studies reported that circulating hormones may affect serum IGF I levels, with potential consequences on bone remodeling. However, no data on bone matrix concentrations of IGF-I in subjects with endocrine dysfunction is available in humans. Bone mineral density and cancellous bone matrix IGF-I levels were assessed in iliac crest biopsies from 38 patients with low bone mass related to glucocorticoid- (n=10), parathyroid- (n=14) or thyroid (n=14) hormones excess. Results were compared to those of sex- and age-matched patients with primary osteoporosis. Bone matrix extraction was performed based on a guanidine chlorhidric acid/ethylendiamine-tetraacetic acid method. Long-term glucocorticoid therapy (> or =24 months) led to significantly lower cancellous bone matrix IGF-I levels in comparison to age-matched controls (p=0.03). Although higher trabecular bone IGF-I levels were seen in hyperparathyroid subjects, the difference was not significant in comparison to controls (p=0.24). Likewise, no difference was noticed in cancellous bone matrix IGF-I concentrations between subjects with low bone mass and sub-clinical or overt thyrotoxicosis and euthyroid controls. Neither parathyroid hormone (PTH) nor thyroxin (T (4)) concentrations were associated with bone matrix IGF-I levels. To conclude, our study documented that in vivo long-term corticotherapy is associated with low trabecular human bone matrix IGF-I. In contrast, no influence of increased circulating parathyroid- or thyroid hormones levels on human iliac crest skeletal IGF-I concentrations was observed. PMID- 19998243 TI - The effect of metformin treatment for 2 years without caloric restriction on endocrine and metabolic parameters in women with polycystic ovary syndrome. AB - INTRODUCTION: The aim of this study was to evaluate long-term changes in endocrine and metabolic parameters and body mass index in women with polycystic ovary syndrome (PCOS) who were treated with metformin over 2 years without caloric restriction. MATERIAL AND METHODS: Twenty-six obese women with PCOS were treated with metformin over 2 years without caloric restriction. Clinical, metabolic and endocrine parameters and the body mass index were measured and an oral glucose tolerance test was carried out to calculate insulin resistance indices at the beginning and at the follow-up after 2 years. The Homeostatic Model for Assessment of Insulin Resistance (HOMA-IR) was calculated. RESULTS: No significant changes in body mass index or HOMA-IR were observed. However, a significant decrease in fasting and 2-h insulin levels was observed. Women showed a significant increase in sex hormone-binding globulin (SHBG) levels, while total testosterone (TT) levels and the free androgen index decreased significantly. Furthermore a significant decrease in hirsutism was observed. There was a decrease in cholesterol and an increase in high-density lipoprotein. CONCLUSIONS: Long-term treatment with metformin in women with PCOS appears to reduce androgen excess due to increased SHBG and decreased TT levels resulting in improvement of hirsutism as a clinical sign of androgen excess. Furthermore a significant decrease in fasting and 2-h insulin levels and slightly improved insulin resistance indices were observed. PMID- 19998244 TI - Urinary iodide excretion in adults in germany 2005 meets WHO target. AB - BACKGROUND: Recent regional and germanwide investigations have shown that the abolition of the requirement to declare iodine in foodstuffs and the greater emphasis on information about goitre prevention led to an increase in urinary iodine excretion in German schoolchildren. There was also a decrease in thyroid size and goitre prevalence in children. No up to date results in adults for the whole of Germany are available. METHODS: In 2005 we examined the urinary iodine excretion in the spontaneous morning urine of 1 538 healthy adults in 357 places from all over Germany. The iodine was measured by the Cer- arsenit method. RESULTS: The median iodine excretion amounted to 132 microg/l. There were no significant differences between age groups, sexes or regions. 64% had no iodine deficiency (>100 microg/l). In 23% the deficiency was slight (50-99 microg/l), in 10% moderate (20-49 microg/l) and in 3% there was severe iodine deficiency (<20 microg/l). 29% excreted >200 microg iodide per litre urine. CONCLUSION: According to the WHO guidelines, there is no longer an iodine deficiency in German adults. PMID- 19998245 TI - Association of the CYP27B1 C(-1260)A polymorphism with autoimmune Addison's disease. AB - Autoimmune Addison's disease (AAD) is a complex endocrine disorder with several susceptibility loci. This study was aimed to investigate the associations of CYP27B1 C(-1260)A and PDCD1 G7146A polymorphisms with AAD in a Polish cohort, comprising 101 AAD patients and 251 healthy controls. CYP27B1 encodes 1alpha hydroxylase, responsible for conversion of the vitamin D (3) precursor into its active form, involved in the immune function. PDCD1 gene gives rise to an inhibitory immune receptor, expressed on activated lymphocytes. Polymorphic variants of these genes had previously been associated with various autoimmune disorders. Genotyping was performed by PCR-RFLP method. The CYP27B1 C(-1260) allele appeared significantly more frequent in AAD compared to controls ( P=0.020), yielding an OR of 1.53 (95% CI 1.07-2.19). The distribution of C( 1260)A genotypes also demonstrated significant difference ( P=0.003). Stratification according to the presence of concomitant autoimmune disorders revealed an association of the C(-1260) allele with the polyendocrine cases of AAD ( P=0.031), while no significance was found for the isolated ADD compared with healthy controls ( P=0.253). Overall, the association between AAD and C( 1260)A was confirmed in a meta-analysis of 325 AAD patients and 952 controls from three different European populations. Under a fixed-effect model, C(-1260) allele and CC genotype were associated with AAD susceptibility with a pooled OR of 1.44 (95% CI 1.18-1.75) and 1.88 (95% CI 1.42-2.36), respectively. No differences were observed for the PDCD1 G7146A between affected subjects and controls (p>0.05). In conclusion, this study confirms the association of the CYP27B1 C(-1260)A polymorphism with AAD, whereas the contribution of PDCD1 G7146A seems less likely. PMID- 19998246 TI - [Where can one tax health care maintenance after the Federal election?]. PMID- 19998247 TI - [Publication price for psychiatry students 2008/2009]. PMID- 19998248 TI - Percutaneous cryoablation techniques and clinical applications. AB - Once requiring surgery, cryoablation now can be performed percutaneously under image guidance, thanks to the development of small probes. Sonography was used to guide cryoablation performed surgically; now, computed tomography and magnetic resonance images are typically used to guide percutaneous cryoablation. Intraprocedural monitoring helps those performing the procedure to treat the tumor completely, while avoiding complications. Percutaneous cryoablation has been shown to be safe and effective for many clinical applications including kidney, liver, prostate, breast, and musculoskeletal cancers. In this article, we briefly review percutaneous cryoablation techniques and clinical applications. PMID- 19998249 TI - Carotid artery stenting: a systematic review of randomized clinical trials. AB - Carotid occlusive disease is responsible for a significant proportion of major adverse cardiovascular events (death, stroke, myocardial infarction). Effective prevention by means of revascularization is a sufficient treatment, if performed at a center with an acceptably low procedural complication rate. Carotid surgery is the currently accepted standard of treatment for revascularization of extra cranial carotid occlusive disease. This has been validated by randomized clinical trials that have demonstrated its efficacy over best medical therapy. However, less invasive protected carotid artery stenting (CAS) has emerged as a potential therapeutic alternative to carotid endarterectomy (CEA) for the treatment of carotid atherosclerotic disease. Over the past decade several clinical trials have compared endovascular with surgical treatment. The Stenting and Angioplasty with Protection in Patients at High Risk for Endarterectomy (SAPPHIRE) trial favored stenting over surgery in high-risk patients. The effectiveness of endovascular treatment in low-risk patients and patients with asymptomatic stenoses at preventing of stroke is still uncertain. Carotid artery stenting with an embolic protection device cannot be considered a scientifically sound and evidence-based alternative to carotid surgery in low-risk and asymptomatic patients until we have the results of further randomized trials. This overview presents the currently available data from randomized trials. PMID- 19998250 TI - Varicose veins and chronic venous insufficiency. AB - Varicose veins are a very frequent disorder with prevalence in our adult population between 14% for large varices and 59% for small teleangiectasias. Subjective symptoms may be very non-specific. The term "chronic venous insufficiency (CVI)" defines functional abnormalities of the venous system producing advanced symptoms like oedema, skin changes or leg ulcers. Both entities, varicose veins and CVI, may be summarized under the term "chronic venous disorders" which includes the full spectrum of morphological and functional abnormalities of the venous system. A classification system to describe chronic venous disorders regarding clinical appearance, etiology, anatomical distribution and pathophysiology has been proposed under the acronym of CEAP. The revised version of the CEAP classification contains also definitions of clinical signs and suggests three levels of apparative investigations adjusted to the clinical stage. Concerning the etiology of venous disorders controversial theories exist leading to different therapeutic concepts. As a matter of fact there is a vicious circle between structural changes in valves and venous wall and hemodynamic forces leading to reflux and venous hypertension. Different methods for treating varicose veins are available producing satisfactory early outcome in most cases, but followed by a high recurrence rate after years. Chronic venous insufficiency requires "chronic management". Compression therapy by bandages for initial treatment of severe stages and maintenance therapy using medical compression stockings is essential. In addition correction of venous refluxes by surgery or endovenous procedures including echo-guided foam sclerotherapy should be considered in every single case. PMID- 19998251 TI - Structured surveillance of patients with peripheral arterial occlusive disease after peripheral vascular interventions. AB - Longterm follow-up and surveillance after either an endovascular or an open surgical vascular intervention is an essential part of the angiologist's or vascular surgeon's care for patients with peripheral arterial disease. This includes diagnosis and treatment of cardiovascular risk factors, selection and monitoring of antiplatelet and/or anticoagulant therapy after revascularization, and surveillance of the results of the vascular intervention. This integrated approach to patients' care will improve patients' prognosis quoad vitam and quoad extremitatem and their quality of life. We summarize the rationale and evidence based practical approaches and guidelines for structured long term surveillance of patients with peripheral arterial disease after a vascular intervention. PMID- 19998252 TI - Surgical reconstructions in peripheral arterial occlusive disease. AB - The prognosis of patients suffering from peripheral arterial occlusive disease (PAD) is directly correlated with the severity of the disease. In critically ischemic legs, after one year only 50% will be alive with a preserved leg. The other 50% will die or undergo an amputation during this time. Reconstructive surgery is highly effective in PAD caused by extensive arterial lesions. Depending on the localization of the occlusion, operative procedures range from local desobliteration to profundoplasty and from aortofemoral to femorodistal bypass procedures. Especially in critical ischemia, time is of the essence for limb salvage. Evidence-based data for diagnosis and operative treatment are described in detail. Only the consequent use of these critical techniques can improve the prognosis of these patients. PMID- 19998253 TI - Surgical treatment of arterial ischemia associated with the use of the angioseal vascular closure device. AB - BACKGROUND: Despite the theoretical effectiveness of the Angio-Seal closure device to control bleeding after arterial puncture it can increase the risk of ischemia. OBJECTIVES: To describe arterial lesions caused by the device, surgical techniques needed to repair those lesions and surgical outcome in patients who underwent surgery for arterial ischemia after heart catheterization. PATIENTS AND METHODS: Seven patients underwent surgery over a period of 12 months at our institution. Five patients underwent emergency surgery and two a delayed procedure. The cause of ischemia was dissection of an atheroma plaque at the puncture site in four cases, dissection of the superficial femoral artery in one case, thrombosis of the common femoral artery in one case, and plication of the posterior arterial wall in one case. Arterial repair in these patients required the insertion of a vascular graft in three cases, endarterectomy plus angioplasty in two cases, endarterectomy plus graft interposition plus thrombectomy of the superficial femoral artery in one case and endarterectomy plus femoropopliteal bypass in one case. RESULTS: Treatment was successful in all patients. Mean follow up was 7.6 months (range 5-11 months). During the study period one patient died due to cardiopathy. No patients had to be re-operated and no limb losses were recorded. All the patients were asymptomatic from a vascular point of view with normal active lives for their age. CONCLUSIONS: Surgical repair is effective, although, generally, it is not restricted to a simple thrombectomy, requiring the use of different arterial repair techniques. PMID- 19998254 TI - Computer and internet use in vascular outpatients--ready for interactive applications? AB - BACKGROUND: Exploring patients' computer and internet use, their expectations and attitudes is mandatory for successful introduction of interactive online health care applications in Angiology. PATIENTS AND METHODS: We included 165 outpatients suffering from peripheral arterial disease (PAD; n = 62) and chronic venous and / or lymphatic disease (CVLD; n = 103) in a cross-sectional-study. Patients answered a paper-based questionnaire. RESULTS: Patients were predominantly female (54.5%). 142 (86.1%) reported regular computer use for 9.7 +/- 5.8 years and 134 (81.2 %) used the internet for 6.2 +/- 3.6 years. CVLD-patients and internet-user were younger and higher educated, resulting in a significant difference in computer and internet use between the disease groups (p < 0.01 for both). Time spent online summed up to 4.3 +/- 2.2 days per week and 1.44 +/- 1.2 hours per day for all internet users without significant differences between the groups. The topics retrieved from the internet covered a wide spectrum and searches for health information were mentioned by 41.2 %. Although confidence in the internet (3.3 +/- 1.1 on a 1-6 Likert scale) and reliability in information retrieved from the internet (3.1 +/- 1.1) were relatively low, health-related issues were of high actual and future interest. 42.8% of the patients were even interested in interactive applications like health educational programs, 37.4% in self-reported assessments and outcome questionnaires and 26.9% in chatforums; 50% demanded access to their medical data on an Internetserver. Compared to older participants those < 50 yrs. used the internet more often for shopping, chatting, and e mailing, but not for health information retrieval and interactive applications. CONCLUSIONS: Computers are commonly used and the internet has been adopted as an important source of information by patients suffering from PAD and CVLD. Besides, the internet offers great potentials and new opportunities for interactive disease (self-)management in angiology. To increase confidence and reliability in the medium a careful introduction and evaluation of these new online applications is mandatory. PMID- 19998255 TI - Long-term outcome of patients with mild intermittent claudication under secondary prevention. AB - BACKGROUND: The known natural history of peripheral arterial disease (PAD) is determined by the generalization of atherosclerosis with resulting high cardiovascular and cerebrovascular morbidity and mortality. The aim of this prospective study was to record all vascular and non-vascular events in patients with mild intermittent claudication (IC) undergoing secondary preventive measures in a 10-year follow-up and to determine the time-points at which systemic localizations of atherosclerotic events develop. PATIENTS AND METHODS: Patients who originally had an isolated Fontaine stage IIa PAD were included in the follow up which involved assessments carried out on an annual basis. The incidence and timepoints of vascular (i.e., myocardial infarction, stroke, critical limb ischemia, vascular death), and nonvascular events (i.e., cancer, non vascular death) were recorded and compared with the known natural history. RESULTS: 534 events (vascular: 433) concerning 109 claudicants (M/F: 88/21; 60.8 +/- 8.8 years; ABI 0.66 +/- 0.11) were recorded over an average follow-up period of 104 months. 25.7% of the claudicants died, 39% due to vascular events, 36% due to cancer disease and 25% due to other events. A deterioration of PAD (n = 108) was the most frequent event after 20 months, followed by angina pectoris (n = 41) and cancer diseases (n = 20) after 42 and 45 months, stroke (n = 19) after 58 months, myocardial infarction (n = 12) after 63 months, and finally critical limb ischemia (n = 27) and amputations (n = 10) after 80 and 114 months (median). 111 revascularizations were carried out. 62.3% of the claudicants developed a polyvascular disease, with 20% in a trivascular territory. CONCLUSIONS: A high vascular comorbidity also develops under secondary prevention as an expression of the continuing generalization of the atherosclerotic process in PAD. The causes for death are determined both by the vascular and the tumor related comorbidity. PMID- 19998256 TI - Association of plaque echostructure and cardiovascular risk factors with symptomatic carotid artery disease. AB - BACKGROUND: Severity of stenosis remains the main factor for assessing risk of stroke in patients with internal carotid artery (ICA) disease. This study was conducted to investigate the association of plaque echostructure and other established and emerging cardiovascular risk factors with symptomatic ICA disease. DESIGN: Cross-sectional study of consecutive patients with significant (> 50%) ICA stenosis. PATIENTS AND METHODS: Carotid plaque echostructure, smoking, hypertension, diabetes mellitus, serum lipoprotein (a), homocysteine, vitamin B12, folate, cholesterol to high-density lipoprotein ratio, triglycerides, C-reactive protein, and the Framingham risk score were assessed in 124 consecutive patients (70 asymptomatic; 54 symptomatic) with significant (> 50%) ICA stenosis. RESULTS: The asymptomatic and symptomatic groups did not differ in terms of gender distribution (p = 0.76) and severity of stenosis (p = 0.62). Echolucent plaques (type 1 and 2) were more predominant in patients with symptomatic disease (p = 0.004, OR = 2.13, 95% CI = 1.26-3.6). Patients with plaques type 1 were relatively younger than those with type 4 (p = 0.02). None of the other factors assessed had any significant association with symptomatic disease and any type of carotid plaque. CONCLUSIONS: Besides the severity of carotid stenosis, the presence of an echolucent plaque appears as an important factor associated with symptomatic ICA disease. Also, young patients are more likely to have an echolucent plaque suggesting an age-related association with plaque maturation. PMID- 19998257 TI - Significance of infection markers and microbiological findings during tissue processing of cryopreserved arterial homografts for the early postoperative course. AB - BACKGROUND: To evaluate homograft implantation for the urgent treatment of vascular infections on the basis of the course of infection using microbiological findings in perioperatively obtained specimens and during homograft processing. PATIENTS AND METHODS: 85 patients were treated with cryopreserved homografts from 2004-2007. The microbiological findings of the decontamination process of homografts in the tissue bank were evaluated. The perioperative infection profile (microorganisms, CRP, leukocytes, body temperature) of the patients was analysed. RESULTS: Complete microbiological and clinical follow-up for the postoperative course was available for 35 patients, who were treated with homografts from the same tissue bank and finally included into this study. 55 cryopreserved homografts were implanted. 35/55 (64%) homografts were positive for microorganisms before decontamination. 3/35 (9%) homografts remained positive after the decontamination. 33 patients were operated for prosthetic graft infection and 2 for an infiltration of a large vessel from neighbouring malignant disease. The most common infection agent was Staphylococcus aureus. Thirty-day mortality was 20% (7/35). Only in 4/35 (11%) patients were the microorganisms of the intraoperative swabs also detected during the postoperative course. The microorganisms were ORSA, Enterococcus faecium, Enterobacter aerogenes and Burkholderia cepacia. The patient with ORSA infection died on POD 11 from multiple organ failure and all other patients recovered. None of the postoperative swabs showed the homograft predecontamination microorganisms. Interestingly, a significant association (P = 0.003) between C-reactive protein increase two weeks after surgery and donor-recipient ABO mismatch was found. CONCLUSIONS: The implantation of homografts following the established decontamination is an alternative urgent therapeutic option in vascular infections with encouraging outcomes. The absence of the predecontamination focus in the postoperative specimens of patients, suggests that the postoperative course and outcomes show no strong relation to potential homograft contamination prior to the decontamination process. PMID- 19998258 TI - Cystic adventitial degeneration: ectopic ganglia from adjacent joint capsules. AB - Cystic adventitial degeneration is a rare non-atherosclerotic cause of peripheral arterial occlusive disease, mainly seen in young men without other evidence of vascular disease. Diagnosis will be established by clinical findings and by ultrasound or angiography and can be treated by excision or enucleation of the affected arterial segment or by percutaneous ultrasound-guided aspiration. However, the etiology of adventitial cysts remains unknown. We report a case of cystic adventitial degeneration showing a connection between the joint capsule and the adventitial cyst, supporting the theory that cystic adventitial degeneration may represent ectopic ganglia from adjacent joint capsules. PMID- 19998259 TI - Successful surgical treatment of Nicolau's syndrome combined with intravenous iloprost. AB - A 4 year-old boy was admitted to our clinic with symptoms of pain and ecchymosis in his right leg and foot after injection of benzathine penicilline. There was a localized gangrenous area at the femoral injection site. Doppler ultrasonography showed no arterial flow in the femoral artery and clear evidence of acute thrombosis of the superficial femoral and popliteal veins. Femoral arterial and venous thrombectomy and fasciotomy were performed immediately. After surgery the boy was treated by Iloprost infusion and enoxaparine. One week later necrotic changes had regressed, fasciotomies were closed and only the distal phalanx of the third toe needed amputation. Early surgical intervention and standard management combined with Iloprost infusion may help in healing the lesions by increasing extremity perfusion and may prevent extremity loss. PMID- 19998260 TI - Cardiovocal syndrome secondary to an aortic pseudoaneurysm. AB - Left recurrent laryngeal nerve palsy characterized by hoarseness due to a cardiovascular disorder, which is termed as cardiovocal syndrome or Ortner's syndrome, is an unusual condition. The syndrome might be associated with diverse cardiovascular diseases. However, it is rarely caused by an aortic pseudoaneurysm. The prominent clinical features of such patients are a history of trauma and the injury to or compression of the aortic isthmus involving the laryngeal nerve. Surgical or interventional treatment is necessary, and recurrent laryngeal nerve palsy is usually expected to recover after the surgical intervention of the aortic pseudoaneurysm. PMID- 19998261 TI - Digital ischemic necrosis in a patient with systemic sclerosis: the role of laser Doppler perfusion imaging. AB - The impairment of microcirculation is a cardinal feature of systemic sclerosis (SSc). Raynaud's phenomenon and SSc can cause painful digital ulcers on the fingers or toes, with resulting complications such as serious infections, gangrene or possible amputation. This case report shows the possible role laser Doppler perfusion imaging (LDPI) could play in diagnosis and follow-up of digital arterial obstructions in SSc patients. LDPI is a non-invasive microvascular imaging technique able to provide maps of the cutaneous blood flow (CBF). We describe here the case of a woman affected by SSc who suddenly presented with pallor and pain; afterwards an area of necrosis became clearly visible at the distal phalanx of the third right finger. The LDPI of the dorsal surface of that finger showed a reduction of CBF. Selective arteriography was performed in order to assess the degree and location of the obstruction before proceeding with adventitiectomy. Following this procedure LDPI showed a normal CBF in the distal phalanx and complete healing of the necrotic area. Consequently, we can state that LDPI alone provides an indirect functional estimation of possible digital artery obstruction. Further, we can assert that the overall effect of the adventitiectomy resulted in an increase of the CBF. PMID- 19998262 TI - Occlusion of the celiac trunk, the inferior mesenteric artery and stenosis of the superior mesenteric artery in peripheral thrombangiitis obliterans. AB - Buerger's disease is an inflammatory occlusive disease which commonly involves medium-sized or smaller vessels of extremities. Mesenteric involvement in Buerger's disease is very rare. It can occur at any time during the course of the disease and presents with acute mesenteric ischaemia. In this study, a case of Buerger's disease with mesenteric involvement diagnosed before the onset of acute mesenteric ischaemia and managed endovascularly is reported. PMID- 19998267 TI - Psychiatric disorders among disaster bereaved: an interview study of individuals directly or not directly exposed to the 2004 tsunami. AB - BACKGROUND: Few studies have explored the long-term mental health consequences of disaster losses in bereaved, either exposed to the disaster themselves or not. This study examined the prevalence and predictors of mental disorders and psychological distress in bereaved individuals either directly or not directly exposed to the 2004 tsunami disaster. METHOD: A cross-sectional study of 111 bereaved Norwegians (32 directly and 79 not directly exposed) was conducted 2 years postdisaster. We used a face-to-face structured clinical interview to diagnose current posttraumatic stress disorder (PTSD) and depression (major depressive disorder, MDD) and a self-report scale to measure prolonged grief disorder (PGD). RESULTS: The prevalence of psychiatric disorders was twice as high among individuals directly exposed to the disaster compared to individuals who were not directly exposed (46.9 vs. 22.8 per 100). The prevalence of disorders among the directly exposed was PTSD (34.4%), MDD (25%), and PGD (23.3%), whereas the prevalence among the not directly exposed was PGD (14.3%), MDD (10.1%), and PTSD (5.2%). The co-occurrence of disorders was higher among the directly exposed (21.9 vs. 5.2%). Low education and loss of a child predicted PGD, whereas direct exposure to the disaster predicted PTSD. All three disorders were independently associated with functional impairment. CONCLUSIONS: The dual burden of direct trauma and loss can inflict a complex set of long-term reactions and mental health problems in bereaved individuals. The relationship between PGD and impaired functioning actualizes the incorporation of PGD in future diagnostic manuals of psychiatric disorders. PMID- 19998268 TI - Comments on "Extremely low frequency electric fields and cancer: assessing the evidence" by Kheifets et al. PMID- 19998269 TI - Mefloquine prescriptions in the presence of contraindications: prevalence among US military personnel deployed to Afghanistan, 2007. AB - PURPOSE: Contraindications to mefloquine use include a history of certain prevalent neuropsychiatric disorders, which are thought to increase the risk of severe adverse events including anxiety, paranoia, depression, hallucinations, psychosis, and possibly suicide. Within the US military, the continued availability and use of mefloquine is subject to administrative policies dating to 2002 that require clinicians to exercise added caution during prescribing. This analysis was performed to quantify the effectiveness of these policies in ensuring health care provider compliance with package insert prescribing guidance. METHODS: A previously identified cohort consisting of 11,725 active duty US military personnel, among whom 1127 (9.6%) had contraindications to mefloquine use identified through medical surveillance and pharmaceutical databases, was examined to identify individuals receiving prescriptions for mefloquine in the 45 days prior to a combat deployment in 2007. RESULTS: Among the 11,725 cohort members, 4505 (38.4% of the cohort) received a prescription for mefloquine. Among the 1127 cohort members with contraindications, 155 (1.3% of the cohort) were prescribed mefloquine, comprising 13.8% of those with contraindications. CONCLUSIONS: Despite the longstanding administrative policies meant to reduce such events, approximately one in seven individuals with neuropsychiatric contraindications received a prescription for mefloquine prior to a recent combat deployment, significantly increasing the risk of subsequent adverse events. Given the prevalence of neuropsychiatric disorders among US military personnel and the continued availability of mefloquine, additional study is recommended to describe and quantify the nature and extent of mefloquine associated adverse events experienced among this group. PMID- 19998270 TI - Sustained dopaminergic response of parkinsonism and depression in POLG-associated parkinsonism. PMID- 19998271 TI - Changing dendritic field size of mouse retinal ganglion cells in early postnatal development. AB - During early postnatal development, dendrites of retinal ganglion cells (RGCs) extend and branch in the inner plexiform layer to establish the adult level of stratification, pattern of branching, and coverage. Many studies have described the branching patterns, transient features, and regulatory factors of stratification of the RGCs. The rate of RGC dendritic field (DF) expansion relative to the growing retina has not been systematically investigated. In this study, we used two methods to examine the relative expansion of RGC DFs. First, we measured the size of RGC DFs and the diameters of the eyeballs at several postnatal stages. We compared the measurements with the RGC DF sizes calculated from difference of the eyeball sizes based on a linear expansion assumption. Second, we used the number of cholinergic amacrine cells (SACs) circumscribed by the DFs of RGCs at corresponding time points as an internal ruler to assess the size of DFs. We found most RGCs exhibit a phase of faster expansion relative to the retina between postnatal day 8 (P8) and P13, followed by a phase of retraction between P13 and adulthood. The morphological alpha cells showed the faster growing phase but not the retraction phase, whereas the morphological ON OFF direction selective ganglion cells expanded in the same pace as the growing retina. These findings indicate different RGCs show different modes of growth, whereas most subtypes exhibit a fast expansion followed by a retraction phase to reach the adult size. PMID- 19998272 TI - Long-term efficacy of tenofovir monotherapy for hepatitis B virus-monoinfected patients after failure of nucleoside/nucleotide analogues. AB - Tenofovir disoproxil fumarate (TDF) has demonstrated high antiviral efficacy in treatment-naive patients with chronic hepatitis B virus (HBV) infection but experience in nucleoside/nucleotide analogue (NA)-experienced patients is limited. In this retrospective multicenter study we therefore assessed the long term efficacy of TDF monotherapy in patients with prior failure or resistance to different NA treatments. Criteria for inclusion were HBV DNA levels >4.0 log(10) copies/mL at the start and a minimum period of TDF therapy for at least 6 months. In all, 131 patients (mean age 42 +/- 12 years, 95 male, 65% hepatitis B e antigen [HBeAg]-positive) were eligible. Pretreatment consisted of either monotherapy with lamivudine (LAM; n = 18), adefovir (ADV; n = 8), and sequential LAM-ADV therapy (n = 73), or add-on combination therapy with both drugs (n = 29). Three patients had failed entecavir therapy. Resistance analysis in 113 of the 131 patients revealed genotypic LAM and ADV resistance in 62% and 19% of patients, respectively. The mean HBV DNA level at TDF baseline was 7.6 +/- 1.5 log(10) copies/mL. The overall cumulative proportion of patients achieving HBV DNA levels <400 copies/mL was 79% after a mean treatment duration of 23 months (range, 6-60). Although LAM resistance did not influence the antiviral efficacy of TDF, the presence of ADV resistance impaired TDF efficacy (100% versus 52% probability of HBV DNA <400 copies/mL, respectively). However, virologic breakthrough was not observed in any of the patients during the entire observation period. Loss of HBeAg occurred in 24% of patients and HBsAg loss occurred in 3%. No significant adverse events were noticed during TDF monotherapy. CONCLUSION: TDF monotherapy induced a potent and long-lasting antiviral response in NA-experienced patients with previous treatment failure. Our data may have implications for current add-on strategies. PMID- 19998274 TI - Highly efficient generation of human hepatocyte-like cells from induced pluripotent stem cells. AB - There exists a worldwide shortage of donor livers available for orthotropic liver transplantation and hepatocyte transplantation therapies. In addition to their therapeutic potential, primary human hepatocytes facilitate the study of molecular and genetic aspects of human hepatic disease and development and provide a platform for drug toxicity screens and identification of novel pharmaceuticals with potential to treat a wide array of metabolic diseases. The demand for human hepatocytes, therefore, heavily outweighs their availability. As an alternative to using donor livers as a source of primary hepatocytes, we explored the possibility of generating patient-specific human hepatocytes from induced pluripotent stem (iPS) cells. CONCLUSION: We demonstrate that mouse iPS cells retain full potential for fetal liver development and describe a procedure that facilitates the efficient generation of highly differentiated human hepatocyte-like cells from iPS cells that display key liver functions and can integrate into the hepatic parenchyma in vivo. PMID- 19998275 TI - Micro-Raman spectroscopy of algae: composition analysis and fluorescence background behavior. AB - Preliminary feasibility studies were performed using Stokes Raman scattering for compositional analysis of algae. Two algal species, Chlorella sorokiniana (UTEX #1230) and Neochloris oleoabundans (UTEX #1185), were chosen for this study. Both species were considered to be candidates for biofuel production. Raman signals due to storage lipids (specifically triglycerides) were clearly identified in the nitrogen-starved C. sorokiniana and N. oleoabundans, but not in their healthy counterparts. On the other hand, signals resulting from the carotenoids were found to be present in all of the samples. Composition mapping was conducted in which Raman spectra were acquired from a dense sequence of locations over a small region of interest. The spectra obtained for the mapping images were filtered for the wavelengths of characteristic peaks that correspond to components of interest (i.e., triglyceride or carotenoid). The locations of the components of interest could be identified by the high intensity areas in the composition maps. Finally, the time evolution of fluorescence background was observed while acquiring Raman signals from the algae. The time dependence of fluorescence background is characterized by a general power law decay interrupted by sudden high intensity fluorescence events. The decreasing trend is likely a result of photo-bleaching of cell pigments due to prolonged intense laser exposure, while the sudden high intensity fluorescence events are not understood. PMID- 19998276 TI - Aerated Shewanella oneidensis in continuously fed bioelectrochemical systems for power and hydrogen production. AB - We studied the effects of aeration of Shewanella oneidensis on potentiostatic current production, hydrogen production in a microbial electrolysis cell, and electric power generation in a microbial fuel cell (MFC). The potentiostatic performance of aerated S. oneidensis was considerably enhanced to a maximum current density of 0.45 A/m(2) or 80.3 A/m(3) (mean: 0.34 A/m(2), 57.2 A/m(3)) compared to anaerobically grown cultures. Biocatalyzed hydrogen production rates with aerated S. oneidensis were studied within the applied potential range of 0.3 0.9 V and were highest at 0.9 V with 0.3 m(3) H(2)/m(3) day, which has been reported for mixed cultures, but is approximately 10 times higher than reported for an anaerobic culture of S. oneidensis. Aerated MFC experiments produced a maximum power density of 3.56 W/m(3) at a 200-Omega external resistor. The main reasons for enhanced electrochemical performance are higher levels of active biomass and more efficient substrate utilization under aerobic conditions. Coulombic efficiencies, however, were greatly reduced due to losses of reducing equivalents to aerobic respiration in the anode chamber. The next challenge will be to optimize the aeration rate of the bacterial culture to balance between maximization of bacterial activation and minimization of aerobic respiration in the culture. PMID- 19998273 TI - Annual report to the nation on the status of cancer, 1975-2006, featuring colorectal cancer trends and impact of interventions (risk factors, screening, and treatment) to reduce future rates. AB - BACKGROUND: The American Cancer Society, the Centers for Disease Control and Prevention (CDC), the National Cancer Institute (NCI), and the North American Association of Central Cancer Registries (NAACCR) collaborate annually to provide updated information regarding cancer occurrence and trends in the United States. This year's report includes trends in colorectal cancer (CRC) incidence and death rates and highlights the use of microsimulation modeling as a tool for interpreting past trends and projecting future trends to assist in cancer control planning and policy decisions. METHODS: Information regarding invasive cancers was obtained from the NCI, CDC, and NAACCR; and information on deaths was obtained from the CDC's National Center for Health Statistics. Annual percentage changes in the age-standardized incidence and death rates (based on the year 2000 US population standard) for all cancers combined and for the top 15 cancers were estimated by joinpoint analysis of long-term trends (1975-2006) and for short term fixed-interval trends (1997-2006). All statistical tests were 2-sided. RESULTS: Both incidence and death rates from all cancers combined significantly declined (P < .05) in the most recent time period for men and women overall and for most racial and ethnic populations. These decreases were driven largely by declines in both incidence and death rates for the 3 most common cancers in men (ie, lung and prostate cancers and CRC) and for 2 of the 3 leading cancers in women (ie, breast cancer and CRC). The long-term trends for lung cancer mortality in women had smaller and smaller increases until 2003, when there was a change to a nonsignificant decline. Microsimulation modeling demonstrates that declines in CRC death rates are consistent with a relatively large contribution from screening and with a smaller but demonstrable impact of risk factor reductions and improved treatments. These declines are projected to continue if risk factor modification, screening, and treatment remain at current rates, but they could be accelerated further with favorable trends in risk factors and higher utilization of screening and optimal treatment. CONCLUSIONS: Although the decrease in overall cancer incidence and death rates is encouraging, rising incidence and mortality for some cancers are of concern. PMID- 19998277 TI - Impact of recycling stillage on conversion of dilute sulfuric acid pretreated corn stover to ethanol. AB - Both the current corn starch to ethanol industry and the emerging lignocellulosic biofuels industry view recycling of spent fermentation broth or stillage as a method to reduce fresh water use. The objective of this study was to understand the impact of recycling stillage on conversion of corn stover to ethanol. Sugars in a dilute-acid pretreated corn stover hydrolysate were fermented to ethanol by the glucose-xylose fermenting bacteria Zymomonas mobilis 8b. Three serial fermentations were performed at two different initial sugar concentrations using either 10% or 25% of the stillage as makeup water for the next fermentation in the series. Serial fermentations were performed to achieve near steady state concentration of inhibitors and other compounds in the corn stover hydrolysate. Little impact on ethanol yields was seen at sugar concentrations equivalent to pretreated corn stover slurry at 15% (w/w) with 10% recycle of the stillage. However, ethanol yields became progressively poorer as the sugar concentration increased and fraction of the stillage recycled increased. At an equivalent corn stover slurry concentration of 20% with 25% recycled stillage the ethanol yield was only 5%. For this microorganism with dilute-acid pretreated corn stover, recycling a large fraction of the stillage had a significant negative impact on fermentation performance. Although this finding is of concern for biochemical based lignocellulose conversion processes, other microorganism/pretreatment technology combinations will likely perform differently. PMID- 19998278 TI - The effect of isolated lignins, obtained from a range of pretreated lignocellulosic substrates, on enzymatic hydrolysis. AB - The influence of the residual lignin remaining in the cellulosic rich component of pretreated lignocellulosic substrates on subsequent enzymatic hydrolysis was assessed. Twelve lignin preparations were isolated by two isolation methods (protease treated lignin (PTL) and cellulolytic enzymatic lignin (CEL)) from three types of biomass (corn stover, poplar, and lodgepole pine) that had been pretreated by two processes (steam and organosolv pretreatments). Comparative analysis of the isolated lignin showed that the CEL contained lower amounts of carbohydrates and protein than did the PTL and that the isolated lignin from corn stover contained more carbohydrates than did the lignin derived from the poplar and lodgepole pine. The lower yields of acid insoluble lignin (AIL) obtained from the corn stover when using the PTL method indicated that the lignin from the corn stover had a higher hydrophilicity than did the lignin from the poplar and lodgepole pine. The isolated lignin preparations were added to the reaction mixture containing crystalline cellulose (Avicel) and their possible effects on enzymatic hydrolysis were assessed. It was apparent that the lignin isolated from lodgepole pine and steam pretreated poplar decreased the hydrolysis yields of Avicel, whereas the other isolated lignins did not appear to decrease the hydrolysis yields significantly. The hydrolysis yields of the pretreated lignocellulose and those of Avicel containing the PTL showed good correlation, indicating that the nature of the residual lignin obtained after pretreatment significantly influenced hydrolysis. PMID- 19998279 TI - A sustainable pathway of cellulosic ethanol production integrating anaerobic digestion with biorefining. AB - Anaerobic digestion (AD) of animal manure is traditionally classified as a treatment to reduce the environmental impacts of odor, pathogens, and excess nutrients associated with animal manure. This report shows that AD also changes the composition of manure fiber and makes it suitable as a cellulosic feedstock for ethanol production. Anaerobically digested manure fiber (AD fiber) contains less hemicellulose (11%) and more cellulose (32%) than raw manure, and has better enzymatic digestibility than switchgrass. Using the optimal dilute alkaline pretreatment (2% sodium hydroxide, 130 degrees C, and 2 h), enzymatic hydrolysis of 10% (dry basis) pretreated AD fiber produces 51 g/L glucose at a conversion rate of 90%. The ethanol fermentation on the hydrolysate has a 72% ethanol yield. The results indicate that 120 million dry tons of cattle manure available annually in the U.S. can generate 63 million dry tons of AD fiber that can produce more than 1.67 billion gallons of ethanol. Integrating AD with biorefining will make significant contribution to the cellulosic ethanol production. PMID- 19998280 TI - Metabolite stress and tolerance in the production of biofuels and chemicals: gene expression-based systems analysis of butanol, butyrate, and acetate stresses in the anaerobe Clostridium acetobutylicum. AB - Metabolite accumulation has pleiotropic, toxic, or beneficial effects on cell physiology, but such effects are not well understood at the molecular level. Cells respond and adapt to metabolite stress by mechanisms largely unexplored, especially in the context of multiple and simultaneous stresses. Solventogenic and related clostridia have an inherent advantage for production of biofuels and chemicals directly from cellulosic material and other complex carbohydrates, but issues of product/metabolite tolerance and related culture productivities remain. Using DNA microarray-based gene expression analysis, the transcriptional-stress responses of Clostridium acetobutylicum to fermentation acids acetate and butyrate and the solvent product butanol were analyzed and compared in the context of cell physiology. Ontological analysis demonstrated that stress by all three metabolites resulted in upregulation of genes related to post-translational modifications and chaperone activity, and downregulation of the translation machinery genes. Motility genes were downregulated by acetate-stress only. The general metabolite stress included upregulation of numerous stress genes (dnaK, groES, groEL, hsp90, hsp18, clpC, and htrA), the solventogenic operon aad-ctfA ctfB, and other solventogenic genes. Acetate stress downregulated expression of the butyryl-CoA- and butyrate-formation genes, while butyrate stress downregulated expression of acetate-formation genes. Pyrimidine-biosynthesis genes were downregulated by most stresses, but purine-biosynthesis genes were upregulated by acetate and butyrate, possibly for thiamine and histidine biosynthesis. Methionine-biosynthesis genes were upregulated by acetate stress, indicating a possibly conserved stress response mechanism also observed in Escherichia coli. Nitrogen-fixation gene expression was upregulated by acetate stress. Butyrate stress upregulated many iron-metabolism genes, riboflavin biosynthesis genes, and several genes related to cellular repair from oxidative stress, such as perR and superoxide dismutases. Butanol stress upregulated the glycerol metabolism genes glpA and glpF. Surprisingly, metabolite stress had no apparent effect on the expression of the sporulation-cascade genes. It is argued that the list of upregulated genes in response to the three metabolite stresses includes several genes whose overexpression would likely impart tolerance, thus making the information generated in this study, a valuable source for the development of tolerant recombinant strains. PMID- 19998281 TI - Production of biocommodities and bioelectricity by cell-free synthetic enzymatic pathway biotransformations: challenges and opportunities. AB - Cell-free synthetic (enzymatic) pathway biotransformation (SyPaB) is the assembly of a number of purified enzymes (usually more than 10) and coenzymes for the production of desired products through complicated biochemical reaction networks that a single enzyme cannot do. Cell-free SyPaB, as compared to microbial fermentation, has several distinctive advantages, such as high product yield, great engineering flexibility, high product titer, and fast reaction rate. Biocommodities (e.g., ethanol, hydrogen, and butanol) are low-value products where costs of feedstock carbohydrates often account for approximately 30-70% of the prices of the products. Therefore, yield of biocommodities is the most important cost factor, and the lowest yields of profitable biofuels are estimated to be ca. 70% of the theoretical yields of sugar-to-biofuels based on sugar prices of ca. US$ 0.18 per kg. The opinion that SyPaB is too costly for producing low-value biocommodities are mainly attributed to the lack of stable standardized building blocks (e.g., enzymes or their complexes), costly labile coenzymes, and replenishment of enzymes and coenzymes. In this perspective, I propose design principles for SyPaB, present several SyPaB examples for generating hydrogen, alcohols, and electricity, and analyze the advantages and limitations of SyPaB. The economical analyses clearly suggest that developments in stable enzymes or their complexes as standardized parts, efficient coenzyme recycling, and use of low-cost and more stable biomimetic coenzyme analogs, would result in much lower production costs than do microbial fermentations because the stabilized enzymes have more than 3 orders of magnitude higher weight-based total turn-over numbers than microbial biocatalysts, although extra costs for enzyme purification and stabilization are spent. PMID- 19998282 TI - Multi-species nitrifying biofilm model (MSNBM) including free ammonia and free nitrous acid inhibition and oxygen limitation. AB - A multi-species nitrifying biofilm model (MSNBM) is developed to describe nitrite accumulation by simultaneous free ammonia (FA) and free nitrous acid (FNA) inhibition, direct pH inhibition, and oxygen limitation in a biofilm. The MSNBM addresses the spatial gradient of pH with biofilm depth and how it induces changes of FA and FNA speciation and inhibition. Simulations using the MSNBM in a completely mixed biofilm reactor show that influent total ammonia nitrogen (TAN) concentration, bulk dissolved oxygen (DO) concentration, and buffer concentration exert significant control on the suppression of nitrite-oxidizing bacteria (NOB) and shortcut biological nitrogen removal (SBNR), but the pH in the bulk liquid has a weaker influence. Ammonium oxidation increases the nitrite concentration and decreases the pH, which together can increase FNA inhibition of NOB in the biofilm. Thus, a low buffer concentration can accentuate SBNR. DO and influent TAN concentrations are efficient means to enhance DO limitation, which affects NOB more than ammonia-oxidizing bacteria (AOB) inside the biofilm. With high influent TAN concentration, FA inhibition is dominant at an early phase, but finally DO limitation becomes more important as TAN degradation and biofilm growth proceed. MSNBM results indicate that oxygen depletion and FNA inhibition throughout the biofilm continuously suppress the growth of NOB, which helps achieve SBNR with a lower TAN concentration than in systems without concentration gradients. PMID- 19998283 TI - Modeling predation processes in activated sludge. AB - Predation by protozoa plays an important role in activated sludge. In this work, the kinetics for protozoan predation of active bacteria (X(H)), extracellular polymeric substances (EPS), and intracellular storage products (X(STO)) are added into a previously expanded unified model that describes the dynamics of EPS, X(STO), and soluble microbial products (SMP). The new biomass growth-decay predation model describes the biomass fractions, soluble organic components, and oxygen-uptake rates considering EPS, X(STO), and predators during dynamic operating conditions in activated sludge. Model calibration using batch experimental data provides the new parameter values for predation processes and insights into mechanisms involving predators. The calibrated value of the maximum specific growth rate for the predators is much slower than for the bacteria, confirming that predators are relatively slow growers. However, the predators and bacteria have similar decay rates and dissolved oxygen affinities. Model testing with results independent of the calibration data shows two things. First, the model and calibrated parameters accurately simulate the independent results when predators are present. Second, eliminating predation by high salinity significantly lowers the OUR, and this is captured by the model. PMID- 19998284 TI - Potential hydrophobic interaction between two cysteines in interior hydrophobic region improves thermostability of a family 11 xylanase from Neocallimastix patriciarum. AB - In this study, we employed directed evolution and site-directed mutagenesis to screen thermostable mutants of a family 11 xylanase from Neocallimastix patriciarum, and found that the thermostability and specific activity are both enhanced when mutations (G201C and C60A) take place in the interior hydrophobic region of the enzyme. Far-ultraviolet circular dichroism analysis showed that the melting temperatures (T(m)) of the G201C and C60A-G201C mutants are higher than that of the wild type by about 10 and 12 degrees C, respectively. At 72 degrees C, their specific activities are about 4 and 6 times as that of the wild type, respectively. Homology modeling and site-directed mutagenesis demonstrated that the enhanced thermostability of the G201C and C60A-G201C mutants may be mainly attributed to a potential stronger hydrophobic interaction between the two well packed cysteines at sites 50 and 201, rather than the disulfide bond formation which was ruled out by thiol titration with dithionitrobenzoic acid (DTNB). And the strength of such interaction depends on the packing of the side-chain and hydrophobicity of residues at these two sites. This suggests that cysteine could stabilize a protein not only by forming a disulfide bond, but also by the strong hydrophobicity itself. PMID- 19998285 TI - Biohydrogen production from wheat straw hydrolysate by dark fermentation using extreme thermophilic mixed culture. AB - Hydrolysate was tested as substrate for hydrogen production by extreme thermophilic mixed culture (70 degrees C) in both batch and continuously fed reactors. Hydrogen was produced at hydrolysate concentrations up to 25% (v/v), while no hydrogen was produced at hydrolysate concentration of 30% (v/v), indicating that hydrolysate at high concentrations was inhibiting the hydrogen fermentation process. In addition, the lag phase for hydrogen production was strongly influenced by the hydrolysate concentration, and was prolonged from approximately 11 h at the hydrolysate concentrations below 20% (v/v) to 38 h at the hydrolysate concentration of 25% (v/v). The maximum hydrogen yield as determined in batch assays was 318.4 +/- 5.2 mL-H(2)/g-sugars (14.2 +/- 0.2 mmol H(2)/g-sugars) at the hydrolysate concentration of 5% (v/v). Continuously fed, and the continuously stirred tank reactor (CSTR), operating at 3 day hydraulic retention time (HRT) and fed with 20% (v/v) hydrolysate could successfully produce hydrogen. The hydrogen yield and production rate were 178.0 +/- 10.1 mL H(2)/g-sugars (7.9 +/- 0.4 mmol H(2)/g-sugars) and 184.0 +/- 10.7 mL-H(2)/day L(reactor) (8.2 +/- 0.5 mmol-H(2)/day L(reactor)), respectively, corresponding to 12% of the chemical oxygen demand (COD) from sugars. Additionally, it was found that toxic compounds, furfural and hydroxymethylfurfural (HMF), contained in the hydrolysate were effectively degraded in the CSTR, and their concentrations were reduced from 50 and 28 mg/L, respectively, to undetectable concentrations in the effluent. Phylogenetic analysis of the mixed culture revealed that members involved hydrogen producers in both batch and CSTR reactors were phylogenetically related to the Caldanaerobacter subteraneus, Thermoanaerobacter subteraneus, and Thermoanaerobacterium thermosaccharolyticum. PMID- 19998286 TI - Enhanced degradation of phenanthrene in a solid-liquid two-phase partitioning bioreactor via sonication. AB - The current article examined the feasibility of inducing improved delivery and degradation of phenanthrene in a solid-liquid partitioning bioreactor system at bench scale by means of ultrasonic energy input. Initial degradation rates of phenanthrene by a microbial consortium, delivered from Desmopan, were improved 2.7-fold in the presence of sonication relative to unsonicated controls. Results demonstrated that an operating window involving on/off sonication cycling improved substrate delivery and rational selection of ultrasound cycling profiles could lead to even further enhancements. Additionally, all results were obtained in a conventional bioreactor with commercial ultrasonic equipment and a commercially available polymer. Subsequent DGGE analysis demonstrated that the sonication cycles selected maintained consortium compositions, relative to control cases, and suggest that exposure would not reduce degradative capabilities under the periods of irradiation examined. Finally, consortium members were identified as belonging to the Pandoraea, Sphingobium, and Pseudoxanthomonas genera. Comparison of genetic sequences in the Ribosomal Database Project revealed that some of the bacterial members, identified at the strain level, had been previously observed in PAH degradations, while others have been reported only in the degradation of other aromatics, such as pesticides. PMID- 19998287 TI - An accumulative site-specific gene integration system using Cre recombinase mediated cassette exchange. AB - The Cre-loxP system is frequently used for site-specific recombination in animal cells. The equilibrium and specificity of the recombination reaction can be controlled using mutated loxPs. In the present study, we designed an accumulative site-specific gene integration system using Cre recombinase and mutated loxPs in which the Cre-mediated cassette exchange reaction is infinitely repeatable for target gene integration into loxP target sites. To evaluate the feasibility and usefulness of this system, a series of integration reactions were repeated and confirmed in vitro using Cre recombinase protein and plasmids. Accumulative gene integration was also performed on the genome of Chinese hamster ovary (CHO) cells. The results indicated that the system was applicable for repeated gene integration of multiple genes to the target sites on both plasmids and CHO cell genomes. This gene integration system provides a novel strategy for gene amplification and for biological analyses of gene function through the genetic modification of cells and organisms. PMID- 19998290 TI - Optical assessment of cutaneous blood volume depends on the vessel size distribution: a computer simulation study. AB - A Monte Carlo simulation was adapted to specify a skin model with pigmented epidermis, dermis with low baseline blood content, and vessels of a vascular lesion with an average added blood volume fraction of 5%. In the study, the lesion vessel diameters were increased and the number of lesion vessels decreased, such that the total lesion blood content was constant. The results show that reflectance (R) increases as vessel size increases, even though the blood content is constant. Light cannot penetrate effectively into larger blood vessels, so the interior of the vessel becomes less visible to the light - a result known in the literature. This study did repeated random placement of vessels to learn the variation in R due to variable vessel placement. The coefficient of variation was about 10% due to random placement. R varies with size, even with small vessels, and does not simply apply to large vessels. PMID- 19998291 TI - Local vibrational mechanism for negative thermal expansion: a combined neutron scattering and first-principles study. PMID- 19998292 TI - Probing in real time the soft crystallization of DNA-capped nanoparticles. PMID- 19998293 TI - Selective aerobic oxidation of benzylic alcohols catalyzed by carbon-based catalysts: a nonmetallic oxidation system. PMID- 19998294 TI - Transition-metal-catalyzed direct arylation of (hetero)arenes by C-H bond cleavage. AB - The area of transition-metal-catalyzed direct arylation through cleavage of C-H bonds has undergone rapid development in recent years, and is becoming an increasingly viable alternative to traditional cross-coupling reactions with organometallic reagents. In particular, palladium and ruthenium catalysts have been described that enable the direct arylation of (hetero)arenes with challenging coupling partners--including electrophilic aryl chlorides and tosylates as well as simple arenes in cross-dehydrogenative arylations. Furthermore, less expensive copper, iron, and nickel complexes were recently shown to be effective for economically attractive direct arylations. PMID- 19998295 TI - Modeling of the 5'-deiodination of thyroxine by iodothyronine deiodinase: chemical corroboration of a selenenyl iodide intermediate. PMID- 19998296 TI - Thermoresponsive chemical connectors based on hybrid nanowire forests. PMID- 19998297 TI - Boryl-based pincer systems: new avenues in boron chemistry. PMID- 19998298 TI - Sodium-ion-selective two-photon fluorescent probe for in vivo imaging. PMID- 19998299 TI - Surface passivation and transfer doping of silicon nanowires. PMID- 19998300 TI - The twisted side chain of antillatoxin is important for potent toxicity: total synthesis and biological evaluation of antillatoxin and analogues. PMID- 19998301 TI - Four-component synthesis of fully substituted 1,2,4-triazoles. PMID- 19998302 TI - Meso-trialkyl-substituted subporphyrins. PMID- 19998303 TI - Total synthesis of spirastrellolide F methyl ester--part 1: Strategic considerations and revised approach to the southern hemisphere. PMID- 19998304 TI - Neisseria meningitidis serogroup B polysialyltransferase: insights into substrate binding. PMID- 19998305 TI - Gas-phase kinetics of chlorosilylene reactions II. ClSiH + C2H4: absolute rate measurements and quantum chemical and RRKM calculations for the prototype pi addition reaction. AB - Time-resolved studies of chlorosilylene, ClSiH, generated by the 193 nm laser flash photolysis of 1-chloro-1-silacyclopent-3-ene, are carried out to obtain rate constants for its bimolecular reaction with ethene, C(2)H(4), in the gas phase. The reaction is studied over the pressure range 0.13-13.3 kPa (with added SF(6)) at five temperatures in the range 296-562 K. The second order rate constants, obtained by extrapolation to the high pressure limits at each temperature, fitted the Arrhenius equation: log(k(infinity)/cm(3) molecule(-1) s( 1)) = (-10.55+/-0.10) + (3.86 +/- 0.70) kJ mol(-1)/RT ln10. The Arrhenius parameters correspond to a loose transition state and the rate constant at room temperature is 43% of that for SiH(2) + C(2)H(4), showing that the deactivating effect of Cl-for-H substitution in the silylene is not large. Quantum chemical calculations of the potential energy surface for this reaction at the G3MP2//B3LYP level show that, as well as 1-chlorosilirane, ethylchlorosilylene is a viable product. The calculations reveal how the added effect of the Cl atom on the divalent state stabilisation of ClSiH influences the course of this reaction. RRKM calculations of the reaction pressure dependence suggest that ethylchlorosilylene should be the main product. The results are compared and contrasted with those of SiH(2) and SiCl(2) with C(2)H(4). PMID- 19998306 TI - Systematic dielectric and NMR study of the ionic liquid 1-alkyl-3-methyl imidazolium. AB - The dynamic behaviors of ionic liquid samples consisting of a series of 1-alkyl-3 methylimidazolium cations and various counteranionic species are investigated systematically over a wide frequency range from 1 MHz to 20 GHz at room temperature using dielectric relaxation (DR) and nuclear magnetic resonance (NMR) spectroscopies. DR spectra for the ionic liquids are reasonably deconvoluted into two or three relaxation modes. The slowest relaxation times are strongly dependent upon sample viscosity and cation size, whereas the relaxation times of other modes are almost independent of these factors. We attribute the two slower relaxation modes to the rotational relaxation modes of the dipolar cations because the correlation times of the cations evaluated using longitudinal relaxation time (T(1) (13)C NMR) measurements corresponded to the dielectric relaxation times. On the other hand, the fastest relaxation mode is presumably related to the inter-ion motions of ion-pairs formed between cationic and anionic species. In the case of the ionic liquid bis(trifluoromethanesulfonyl)imide, the system shows marked dielectric relaxation behavior due to rotational motion of dipolar anionic species in addition to the relaxation modes attributed to the dipolar cations. PMID- 19998307 TI - Conductivity of poly(pyrrole)-poly(styrene sulfonate) core-shell nanoparticles. AB - The dielectric properties of poly(styrene) nanoparticles decorated at their surfaces with poly(styrene sulfonate) [PSS] brushes and subsequently loaded with polypyrrole (PPy) were studied. These film-forming materials which may serve as hole-injection layers in organic light-emitting diodes, exhibit a core-shell-type morphology with a core of electrically insulating poly(styrene) and a shell consisting of a corona of PSS chains which form the matrix in which the electrically conducting complex of PPy and PSS is embedded. This conducting complex exists in form of domains of nanoscale dimensions. Thin compressed pellets of these nanoparticles were studied using mainly impedance spectroscopy. Measurements were carried out in the temperature range between 123 and 453 K and frequency range from 10(-1) to 10(6) Hz. While earlier studies were centered around the effect of polypyrrole volume fraction on the conductivity films and pellets composed of these nanoparticles, the present study reveals in which way the conductivity can be modified by exchange of the mobile inorganic counter ions of PSS. Besides the free-acid form (H(+)), the Li(+)-, Na(+)- and Cs(+)-salts of PSS were investigated. The PPy volume fraction was the same for all PPy/PSS core shell nanoparticles. The distance for phonon-assisted hopping between next neighbor polypyrrolium chains is influenced by the presence of these inorganic cations. For all samples containing PPy, a transition from insulating to conducting behavior in the range of 300-350 K was found. Using the fluctuation induced tunneling model, the average tunneling distance, as well as the potential energy barrier separating neighboring conducting grains was estimated. Finally, a detailed analysis of the dielectric spectra suggests the localization length of the charge carriers to be about 0.33 nm. PMID- 19998308 TI - Total synthesis of spirastrellolide F methyl ester--part 2: Macrocyclization and completion of the synthesis. PMID- 19998309 TI - Depletion of the ozone layer in the 21st century. PMID- 19998310 TI - Understanding fluorine effects in liquid crystals. PMID- 19998311 TI - Polarizable atomistic calculation of site energy disorder in amorphous Alq3. AB - A polarizable molecular dynamics simulation and calculation scheme for site energy disorder is presented in amorphous tris(8-hydroxyquinolinato)aluminum (Alq(3)) by means of the charge response kernel (CRK) method. The CRK fit to the electrostatic potential and the tight-binding approximation are introduced, which enables modeling of the polarizable electrostatic interaction for a large molecule systematically from an ab initio calculation. The site energy disorder for electron and hole transfers is calculated in amorphous Alq(3) and the effect of the polarization on the site energy disorder is discussed. PMID- 19998312 TI - Mechanism of charging of Au atoms and nanoclusters on Li doped SiO2/Mo(112) films. AB - We present the results of supercell DFT calculations on the adsorption properties of Au atoms and small clusters (Au(n), n < or = 5) on a SiO(2)/Mo(112) thin film and on the same system modified by doping with Li atoms. The adsorbed Li atoms penetrate into the pores of the silica film and become stabilized at the interface where they donate one electron to the Mo metal. As a consequence, the work function of the Li-doped SiO(2)/Mo(112) film is reduced and results in modified adsorption properties. In fact, while on the undoped SiO(2)/Mo(112) film Au interacts only very weakly, on the Li-doped surface Au atoms and clusters bind with significant bond strengths. The calculations show that this is due to the occurrence of an electron transfer from the SiO(2)/Mo(112) interface to the adsorbed gold. The occurrence of the charge transfer is related to the work function of the support but also to the possibility for the silica film to undergo a strong polaronic distortion. PMID- 19998313 TI - Bioinspired peony-like beta-Ni(OH)2 nanostructures with enhanced electrochemical activity and superhydrophobicity. AB - Constructing complex nanostructures has become increasingly important in the development of hydrogen storage, self-cleaning materials, and the formation of chiral branched nanowires. Several approaches have been developed to generate complex nanostructures, which have led to novel applications. Combining biology and nanotechnology through the utilization of biomolecules to chemically template the growth of complex nanostructures during synthesis has aroused great interest. Herein, we use a biomolecule-assisted hydrothermal method to synthesize beta phase Ni(OH)(2) peony-like complex nanostructures with second-order structure nanoplate structure. The novel beta-Ni(OH)(2) nanostructures exhibit high-power Ni/MH battery performance, close to the theoretical capacity of Ni(OH)(2), as well as controlled wetting behavior. We demonstrate that this bioinspired route to generate a complex nanostructure has applications in environmental protection and green secondary cells. This approach opens up opportunities for the synthesis and potential applications of new kinds of nanostructures. PMID- 19998314 TI - Congenital abnormalities in children of 43 pregnant women who attempted suicide with large doses of nitrazepam. AB - OBJECTIVE: To evaluate the teratogenic effect of very large doses of nitrazepam in children born to pregnant women who attempted suicide and to check the feasibility of self-poisoning pregnant women model. DESIGN AND SETTING: Comparative analysis of exposed children and their unexposed sibs born to the same mothers who attempted suicide during the study pregnancy and admitted to the toxicological inpatients clinic, Budapest, 1960-1993. STUDY PARTICIPANTS: Of 1044 pregnant women who attempted suicide, 107 (10.3%) used large doses of nitrazepam alone or combination with other drugs, and 43 delivered live-born babies, these exposed children were evaluated. MAIN OUTCOMES MEASURES: Structural birth defects, i.e., congenital abnormalities (CAs), pregnancy age at delivery, and birth weight. RESULTS: The mean dose of nitrazepam used for suicide attempt was 204 mg. Of 43 exposed children, 13 (30.2%) were affected with CAs, while of their 29 sib controls, 3 (10.3%) (OR with 95%CI: 3.8, 1.0-14.6). Most CAs in exposed children were mild and belonged to the deformation type. The mean pregnancy age was shorter. CONCLUSIONS: The very large doses of nitrazepam used for suicide attempt during pregnancy resulted in a high rate of CAs which may be connected with the disruption of protein metabolism in fetal mesenchyma. The self-poisoning pregnant women model is feasible for the evaluation of teratogenic effect of drugs. PMID- 19998315 TI - The reliability of three depression rating scales in a general population of Dutch older persons. AB - OBJECTIVE: To compare the reliability of three rating scales for assessing depressive symptoms in a community-based, non-clinically depressed older population. METHODS: The study sample comprised of 302 independently living subjects aged 65 years or older. Depressive symptoms were assessed using the centre for epidemiologic studies depression scale (CES-D), the geriatric depression scale (GDS-15) and the Montgomery and Asberg depression rating scale (MADRS) at three time points: at baseline, after 13 weeks (except the GDS-15) and after 26 weeks. Three dimensions of reliability were compared: (i) internal consistency (Cronbach's alpha), (ii) reproducibility (Spearman correlations) and (iii) the intra- and inter-rater reliability (Spearman correlations to compare the differences between correlations of subjects tested by the same vs. different raters at three time points). RESULTS: Cronbach's alpha was high for the CES-D (0.84), good for the MADRS (0.72) and relatively low for the GDS-15 (0.55). Reproducibility was also higher for the CES-D (0.71) than for the MADRS (0.61) and the GDS-15 (0.52). The rater had little influence on CES-D scores (intra/inter-rater ratio = 0.99). The GDS-15 and the MADRS, however, performed better when administered by the same rater. CONCLUSIONS: The CES-D was the most reliable scale for measuring depressive symptoms in a non-clinically depressed older population. PMID- 19998316 TI - Late life depression: a comparison of risk factors and symptoms according to age of onset in community dwelling older adults. AB - BACKGROUND: It has been reported that late onset depression is more frequently associated with acquired organic pathology and that patients are less likely to report a family history of depression. Differences in phenomenology according to age of onset have been described although these have not been consistently replicated. The majority of these studies have been in hospital populations. The aim of this study is to address this question in a sample of community dwelling older adults. METHODS: 89 subjects with GMS-AGECAT depression were identified from a sample of 1231 community dwelling adults aged 65 years and over. Subjects were analysed across a range of aetiological and phenomenological variables according to age of onset of first depressive episode. RESULTS: Subjects with late onset depression (>= 60) were significantly less likely to report a family history of depression, were less likely to report previous hospitalisation for depression and had greater cognitive impairment. Late onset subjects were also less likely to report feelings of guilt or thoughts that life was not worth living in the previous month. CONCLUSION: While we found that patients with late onset depression differed from early onset patients according to certain aetiological risk factors, we did not find a distinctive profile of depressive symptomatology which might be considered clinically useful at an individual level. These findings are consistent with studies based in hospital populations. PMID- 19998317 TI - How chondrogenic are human umbilical cord matrix cells? A comparison to adipose derived stem cells. AB - The umbilical cord matrix as well as liposuction material have been demonstrated to contain cells capable of differentiating towards the mesodermal lineage. High availability and low donor site morbidity appear promising for the use of human umbilical cord matrix cells (HUCMs) and adipose-derived stem cells (ASCs) in cell based therapies. In the present study we focused on cartilage regeneration and compared HUCMs and ASCs regarding their potential to differentiate towards the chondrogenic lineage. Cells were isolated by explantation culture or enzymatic digestion, phenotypically characterized by flow cytometry and differentiated as 3D micromass pellets for up to 35 days. Under tested conditions, ASCs demonstrated significantly higher glycosaminoglycan synthesis compared to HUCMs. qRT-PCR data gave evidence that chondrogenic genes are expressed by both ASCs and HUCMs. However, higher expression levels of ASCs suggest that this cell type has higher potential for differentiation towards a cartilage-like phenotype than HUCMs. In conclusion, both cell types, HUCMs and ASCs, are easily available, possess typical properties of mesenchymal stem cells and are thus promising for cell-based therapies. However, in terms of cartilage regeneration, ASCs might be more suitable than HUCMs. PMID- 19998318 TI - Thiazolidinediones and the risk of incident strokes in patients with type 2 diabetes: a nested case-control study. AB - PURPOSE: To determine whether the use of thiazolidinediones (TZDs) decreases the risk of incident strokes in patients with type 2 diabetes. METHODS: We conducted a nested case-control study within a population-based cohort from the UK General Practice Research Database (GPRD). The cohort comprised patients over the age of 40 who were prescribed a first oral hypoglycemic agent between 1 January 1988 and 30 June 2008. Cases included all subjects who experienced a first stroke during follow-up. Up to 10 controls were matched to each case on age (+/-2 years), sex, date of cohort entry (+/-1 year), and duration of follow-up. Rate ratios (RRs) of stroke associated with TZD use, including rosiglitazone and pioglitazone, relative to combinations of other oral hypoglycemic agents, were estimated using conditional logistic regression. RESULTS: The cohort comprised 75 717 users of oral hypoglycemic agents, of whom 2417 had a stroke during follow-up. The rate of stroke in users of TZDs given as monotherapy (RR: 1.20, 95%CI: 0.77, 1.86) or in combination with other oral hypoglycemic agents (RR: 0.78, 95%CI: 0.58, 1.04) was not lower than combinations of other oral hypoglycemic agents. The RRs were similar for rosiglitazone and pioglitazone. CONCLUSIONS: The results of this study indicate that TZDs do not appear to decrease the incidence of first strokes. PMID- 19998319 TI - How to quantify iron in an aqueous or biological matrix: a technical note. AB - Iron oxide (nano)particles are powerful contrast agents for MRI and tags for magnetic cellular labeling. The need for quantitative methods to evaluate the iron content of contrast media solutions and biological matrixes is thus obvious. Several convenient methods aiming at the quantification of iron from iron oxide nanoparticle-containing samples are presented. PMID- 19998320 TI - How to perform accurate and reliable measurements of longitudinal and transverse relaxation times of MRI contrast media in aqueous solutions. AB - Knowledge of the longitudinal and transverse relaxation times (T(1) and T(2)) of water protons in an aqueous solution of an MRI contrast agent is essential for its characterization. These parameters can be measured at low field on low resolution spectrometers or at high field on high resolution spectrometers. The reliability and the accuracy of T(1) and T(2) measurements rely on several experimental settings and on the equation used to fit the data. Examples of the importance of careful adjustment of the most important parameters are illustrated through several measurements performed on a low-resolution, low-magnetic field instrument. In addition, some specificities of T(1) and T(2) measurements on high resolution, high-magnetic field spectrometers are pointed out. PMID- 19998321 TI - Breath carbon monoxide and semiquantitative saliva cotinine as biomarkers for smoking. AB - OBJECTIVE: As a biomarker of smoking, semiquantitative analysis of cotinine (NicAlert) offers several advantages over breath carbon monoxide (CO) and quantitative analysis of cotinine. Recent studies have used urine NicAlert and breath CO in combination to verify abstinence. However, no studies have evaluated the performance of saliva NicAlert against or in combination with breath CO. METHOD: Breath CO, saliva NicAlert, and smoking history were compared in an urban population of daily smokers (n = 24) and nonsmokers (n = 25). RESULTS: Saliva NicAlert predicted self-reported smoking with 100% sensitivity and 96% specificity. At a cutoff of > 5 ppm, breath CO had 100% sensitivity and 100% specificity in predicting self-reported smoking. Breath CO was positively correlated with saliva NicAlert and negatively correlated with minutes since last cigarette. CONCLUSION: Saliva NicAlert had high sensitivity and specificity in identifying daily smokers. Compared to saliva NicAlert, breath CO level was more indicative of recent smoking. Future treatment studies should evaluate the performance of saliva NicAlert as an alternative to the urine test. PMID- 19998322 TI - Bioguided isolation of angiotensin-converting enzyme inhibitors from the seeds of Plantago asiatica L. AB - Ethanolic extract of the seeds of Plantago asiatica L. showed significant inhibitory activity of angiotensin-converting enzyme (ACE) determined by monitoring the transformation from a substrate hippuryl-histidyl-leucine (HHL) to the product hippuric acid (HA) in vitro using an UPLC-MS method. The bioguided fractionation of the extract resulted in the isolation of four ACE inhibitory active phenylpropanoid glycosides acteoside, isoacteoside, plantainoside D, and plantamajoside with IC(50) values of 2.69 mM, 2.46 mM, 2.17 mM, and 2.47 mM, respectively. Their structures were elucidated through the analysis of NMR, UV, IR and MS data. Our study is the first demonstration that Plantago asiatica L. and its major constituents have ACE inhibitory activity in vitro. It is assumed that the identified compounds contribute to the angiotensin-converting enzyme inhibitory activity of the extract. PMID- 19998323 TI - Berberine: a plant alkaloid with therapeutic potential for central nervous system disorders. AB - Berberine, an isoquinoline alkaloid of the protoberberine type found in an array of plants, has been used in Indian and Chinese medicines as an antimicrobial, stomachic, bitter tonic and in the treatment of oriental sores. Although pharmacological investigations of berberine have been reported by many in the past, there is renewed interest in berberine because of its reported beneficial effect in various neurodegenerative and neuropsychiatric disorders. The alkaloid is reported to modulate neurotransmitters and their receptor systems in the brain. This review attempts to discuss the pharmacological basis of the use of berberine in various central nervous system and related disorders. Its protective effect in Alzheimer's, cerebral ischemia, mental depression, schizophrenia and anxiety are highlighted. However, more detailed clinical trials along with a safety assessment of berberine are warranted for positioning the alkaloid in the treatment of neurological disorders. PMID- 19998324 TI - Pharmacology and immunological actions of a herbal medicine ASHMI on allergic asthma. AB - Allergic asthma is a chronic and progressive inflammatory disease for which there is no satisfactory treatment. Studies reported tolerability and efficacy of an anti-asthma herbal medicine intervention (ASHMI) for asthma patients, developed from traditional Chinese medicine. To investigate the pharmacological actions of ASHMI on early- and late-phase airway responses (EAR and LAR), Ovalbumin (OVA) sensitized mice received 6 weeks of ASHMI treatment beginning 24 h following the first intratracheal OVA challenge. EAR were determined 30 min following the fourth challenge and LAR 48 h following the last challenge. ASHMI effects on cytokine secretion, murine tracheal ring contraction and human bronchial smooth muscle cell prostaglandin (PG) production were also determined.ASHMI abolished EAR, which was associated with significantly reduced histamine, leukotriene C4, and OVA-specific IgE levels, as well as LAR, which was associated with significantly reduced bronchoalveolar lavage fluid (BALF) eosinophils, decreased airway remodeling, and lower Th2 cytokine levels in BALF and splenocyte cultures. Furthermore, ASHMI inhibited contraction of murine tracheal rings and increased production of the potent smooth muscle relaxer PGI(2). ASHMI abrogation of allergic airway responses is associated with broad effects on asthma pathological mechanisms. PMID- 19998325 TI - Fermented ginseng protects streptozotocin-induced damage in rat pancreas by inhibiting nuclear factor-kappaB. AB - In this study, we investigated the protective effects of fermented ginseng (FG) on hyperglycemia induced by streptozotocin (STZ) in Sprague Dawley rats. FG was administered orally at dose of 250 (FGL) or 500 mg/kg (FGH) for 20 days starting one week before STZ injection. FG restored the plasma insulin levels by 266% and 334% in FGL and FGH, respectively, and resulting in reduction of plasma glucose concentration. Histological observation indicated that STZ-induced destruction of pancreatic islets was protected by FG. Consistent with this observation, FG reduced protein and mRNA levels of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2), as determined by Western blotting and RT-PCR, respectively. The molecular mechanism of FG's inhibition of iNOS and COX-2 gene expressions appeared to involve the inhibition of nuclear factor-kappaB (NF kappaB) activation via prevention of inhibitor kappaB (IkappaB) phosphorylation and degradation. The cytoprotective effects of FG were also mediated through suppression of extracelluar signal-regulated kinase (ERK) and c-JUN N-terminal kinase (JNK) pathways. Collectively, these results suggest that FG might be used to preserve functional beta-cell mass. PMID- 19998328 TI - Passaged human chondrocytes accumulate extracellular matrix when induced by bovine chondrocytes. AB - A source of sufficient number of cells is a major limiting factor for cartilage tissue engineering. To circumvent this problem, we developed a co-culture method to induce redifferentiation in bovine articular chondrocytes, which had undergone dedifferentiation following serial passage in monolayer culture. In this study we determine whether human osteoarthritic (OA) and non-diseased passaged dedifferentiated chondrocytes will respond similarly. Human passaged chondrocytes were co-cultured for 4 weeks with primary bovine chondrocytes and their redifferentiation status was determined. Afterwards the cells were cultured either independently or in co-culture with cryopreserved passaged cells for functional analysis. The co-culture of passaged cells with primary chondrocytes resulted in reversion of their phenotype towards articular chondrocytes, as shown by increased gene expression of type II collagen and COMP, decreased type I collagen expression and extracellular matrix formation in vitro. Furthermore, this redifferentiation was stable, as those cells not only formed hyaline-like cartilage tissue when grown on their own but also they could induce redifferentiation of passaged chondrocytes in co-culture. These data suggest that it may be possible to use autologous chondrocytes obtained from osteoarthritic cartilage to form tissue suitable to use for cartilage repair. PMID- 19998329 TI - Cytochrome P450-dependent toxicity of dapsone in human erythrocytes. AB - The most prominent adverse effects seen during treatment with dapsone, an antibacterial and antiprotozoal agent, are hemolysis and methemoglobinemia. An in vitro microsomal/cytochrome P(450) (CYP)-linked assay, which allows reactive metabolites generated in situ to react with the co-incubated human erythrocytes, was employed to profile CYP isoforms responsible for hemotoxicity of dapsone. Dapsone caused a robust generation of methemoglobin in human erythrocytes in the presence of human/mouse liver microsomes, which indicates contribution of CYP mediated metabolism for hemotoxicity. The highest methemoglobin formation with dapsone was observed with CYP2C19, with minor contributions from CYP2B6, CYP2D6 and CYP3A4. Cimetidine and chloramphenicol completely abrogated methemoglobin generation by dapsone, thus confirming a predominant contribution of CYP2C19. The results provide useful insights into CYP-dependent hemotoxicity of dapsone in human erythrocytes. PMID- 19998331 TI - Coping with parental cancer: web-based peer support in children. AB - OBJECTIVE: To investigate the use and content of web-based peer support in children coping with parental cancer. METHODS: In children aged above 12 years, 158 forum discussions on the Dutch website www.kankerspoken.nl in a time-period of 3 months were investigated. Age, gender, number of discussions, participation and main activities on the website were quantitatively assessed. Contents of the discussions were qualitatively analyzed. RESULTS: In total, 129 children and 8 adults participated on the website with an average age of 15.3 years. The majority was female (80%). On average, a child was on-line for 3 days and participated in 4 to 5 forum discussions. Main activities on the website were sharing personal experience, providing encouragement/support, providing and seeking information or advice and seeking contact outside of the website. Qualitative content analysis revealed that children regularly faced emotional problems and experienced a lack of understanding and communication in their direct environment. CONCLUSIONS: In dealing with parental cancer, children experience a variety of difficulties. This study offers a window into the use of web-based peer support by children dealing with parental cancer. PMID- 19998330 TI - Micropatterned three-dimensional hydrogel system to study human endothelial mesenchymal stem cell interactions. AB - The creation of vascularized engineered tissues of clinically relevant size is a major challenge of tissue engineering. While it is known that endothelial and mural vascular cells are integral to the formation of stable blood vessels, the specific cell types and optimal conditions for engineered vascular networks are poorly understood. To this end, we investigated the vasculogenic potential of human mesenchymal stem cell (MSC) populations derived from three different sources: (a) bone marrow aspirates; (b) perivascular cells from the umbilical cord vein; and (c) perivascular cells from the umbilical cord artery. Cell populations were isolated and identified as MSCs according to their phenotypes and differentiation potential. Human umbilical vein endothelial cells (HUVECs) were used as a standard for endothelial cells. A novel co-culture system was developed to study cell-cell interactions in a spatially controlled three dimensional (3D) fibrin hydrogel model. Using microfluidic patterning, it was possible to localize hydrogel-encapsulated HUVECs and MSCs within separate channels spaced at 500, 1000 or 2000 microm. All three MSC populations had similar expression profiles of mesenchymal cell markers and similar capacity for osteogenic and adipogenic differentiation. However, bone marrow-derived MSCs (but not umbilical vein or artery derived MSCs) showed strong distance-dependent migration toward HUVECs and supported the formation of stable vascular networks resembling capillary-like vasculature. The presented approach provides a simple and robust model to study the cell-cell communication of relevance to engineering vascularized tissues. PMID- 19998332 TI - Are cardiovascular diseases bad for economic growth? AB - We assess the impact of cardiovascular disease (CVD) mortality on economic growth, using a dynamic panel growth regression framework taking into account potential endogeneity problems. In the worldwide sample we detect a non-linear influence of working age CVD mortality rates on growth across the per capita income scale. Splitting the sample (according to the resulting income threshold) into low- and middle-income countries, and high-income countries, we find a robust negative contribution of increasing CVD mortality rates on subsequent five year growth rates in the latter sample. Not too surprisingly, we find no significant impact in the low- and middle-income country sample. PMID- 19998333 TI - An evaluation of a computer-imaging program to prepare women for chemotherapy related alopecia. AB - OBJECTIVE: This study was conducted to evaluate a computer program named Help with Adjustment to Alopecia by Image Recovery (HAAIR) that was developed to provide educational support and reduce distress in women with hair loss following chemotherapy. METHODS: Forty-five women who had been diagnosed with cancer and anticipated alopecia following treatment were randomly assigned to either the Imagining group (IG) or Standardized Care group (SCG). Patients in the IG used a computer-imaging program that created the patient's image on a screen to simulate baldness and use of wigs whereas patients in the SCG were directed to a resource room at the Cancer Center established for women with chemotherapy-related alopecia. Assessment data using the Brief Symptom Inventory, Importance of Hair Questionnaire, and the Brief Cope were completed at baseline (T1), before chemotherapy and hair loss, following hair loss (T2), and 3 months follow-up (T3). RESULTS: All women were able to successfully use the touch screen computerized-imaging program and reported that using the computer was a positive, helpful experience, thus establishing acceptability and usability. Women in both the IG and the SCG group showed significantly lower hair loss distress scores at T2 after hair loss than at T1 with T3 distress scores increasing in the SCG and decreasing in the IG. Those with avoidance coping reported more distress. CONCLUSIONS: This evaluation demonstrates that the HAAIR program is a patient endorsed educational and supportive complement to care for women facing chemotherapy-related alopecia. PMID- 19998334 TI - Ras inhibits endoplasmic reticulum stress in human cancer cells with amplified Myc. AB - In neuroblastoma LAN-1 cells harboring an amplified MycN gene, disruption of cooperation between Ras and MycN proteins by the Ras inhibitor farnesylthiosalicylic acid (FTS, Salirasib) reportedly arrests cell growth. Our aim was to establish whether this is a general phenomenon. We examined the effects of FTS on gene-expression profiles, growth and death of NCIH929 myeloma cells and K562 leukemia cells, which-like LAN-1 cells-exhibit Myc gene amplification and harbor active Ras. Under specified conditions, FTS reduced Ras and Myc and induced cell growth arrest and death in all Myc-amplified cell lines but not in SHEP, a neuroblastoma cell line without Myc gene amplification. Gene expression analysis revealed a common pattern of FTS-induced endoplasmic reticulum (ER) stress, known as the unfolded protein response (UPR), in Myc amplified cells, but not in SHEP. Thus, Ras negatively regulates ER stress in cells with amplified Myc. ER stress was also inducible by dominant-negative (DN) Ras or shRNA to Ras isoforms, all of which induced an increase in BIP (the master regulator of ER stress) and its downstream targets Nrf2 and eIF2alpha, both regulated by active p-PERK. FTS also induced an increase in p-PERK, while small interfering RNA to PERK reduced Nrf2 and ATF4 and rescued cells from FTS-induced death. BIP and its downstream targets were also increased by inhibitors of MAPK p38 and MEK. Ras, acting through MAPK p38 and MEK, negatively regulates the ER stress cascades BIP/PERK/Nrf2 and eIF2alpha/ATF4/ATF3. These findings can explain the Ras-dependent protection of Myc-amplified cells from ER stress-associated death. PMID- 19998335 TI - Changes in the risk of death from cancer up to five years after diagnosis in elderly patients: a study of five common cancers. AB - Cancer mortality in elderly people is described to highlight the mechanisms that could potentially explain observed differences with other age groups. Data from 15 French cancer registries were considered in the search for the 5-year outcome of patients diagnosed during the period 1989-1997. Relative survival, excess mortality hazard, and hazard ratio of mortality were estimated to describe patient outcome according to age. Five cancer sites were selected: colon/rectum, prostate, breast, head/neck, and lung. An excess mortality rate was found in patients aged over 75 at the time of diagnosis. This excess mortality rate was mainly seen during the first months after diagnosis, then it decreased gradually with time. An initial phenomenon of patient selection, a greater disease severity at the time of diagnosis, and less-effective treatments given to elderly patients are the most plausible explanations for the increased risk of cancer-related death in the eldest patients. PMID- 19998337 TI - Prognostic significance and therapeutic potential of eukaryotic translation initiation factor 5A (eIF5A) in hepatocellular carcinoma. AB - Using comparative proteomic and genomic approaches, the authors identified eukaryotic translation initiation factor 5A (eIF5A) as an oncofetal molecule highly abundant in mouse embryonic livers and human hepatocellular carcinoma (HCC) cell lines. To evaluate the oncogenic role and prognostic significance of eIF5A in HCC, we investigate the expression patterns of the two isoforms (eIF5A1 and eIF5A2) in a cohort of 258 HCC cases by cDNA microarray. Both eIF5A isoforms were expressed in the tumors, and clinically correlated eIF5A1 with more numbers of tumor nodules and eIF5A2 with tumor venous infiltration in HCC. In a separate cohort of 50 HCCs, high level of eIF5A2, but not eIF5A1, was associated with elevated levels of deoxyhypusine synthase and deoxyhypusine hydroxylase that catalyze post-translational hypusination of eIF5A protein. Interestingly, N1 guanyl-1,7-diaminoheptane (GC7), which is an inhibitor for the first step of eIF5A hypusination, was shown to significantly impair the cell proliferation and invasion of primary HCC cells (HepG2 and Hep3B). To further demonstrate the tumorigenic role associated with eIF5A, a drastic reduction of cell proliferation was associated with suppression of eIF5A2 by transfecting Hep3B, H2-P and H2-M HCC cells expressing high level of this isoform using small interfering RNA (siRNA) against eIF5A2. For these assays, a milder response was usually observed in normal hepatocyte cell line. Therefore, these findings suggest that eIF5A plays an important role in HCC tumorigenesis and metastasis, and targeting eIF5A hypusination by GC7 inhibitor or eIF5A2 by RNA interference (RNAi) may offer new therapeutic alternatives to HCC patients. PMID- 19998336 TI - Dietary fatty acids, luminal modifiers, and risk of colorectal cancer. AB - Inconsistent observations in epidemiologic studies on the association between total fat intake and colorectal cancer may be ascribed to opposing effects of individual fatty acids and the presence of other dietary constituents that modify luminal or systemic lipid exposure. We analyzed the data from a population-based case-control study that included 1,163 cases and 1,501 controls to examine the effects of individual fatty acid groups on colorectal cancer risk as well as their interactions with calcium and fiber intake. Odds ratios (OR) and 95% confidence intervals (CI) were estimated by unconditional logistic regression model according to quartile levels of energy-adjusted fatty acid intake. In the bivariable analyses, the risk of colorectal cancer increased with trans fatty acid (TFA) intake (OR for top vs. bottom quartile =1.46, 95% CI 1.17-1.59, p value for a trend <0.001), but the associations was substantially attenuated in multivariable analyses (p value for a trend = 0.176). However, a significant linear trend in the multivariable OR (p = 0.029) for TFA was present for subjects with lower calcium intake. Furthermore, multivariable ORs progressively decreased with increasing both omega-3 and omega-6 poly- unsaturated fatty acid intake (p values for linear trend: 0.033 and 0.011, respectively) for subjects with lower dietary fiber intake. These interactions were also significant or marginally significant (p = 0.085 for TFA, 0.029 for omega-3 and 0.068 for omega-6). Our results suggest that populations with lower intake of luminal modifiers, i.e., calcium and fiber, may have differential risks of colorectal cancer associated with dietary fatty acid intake. PMID- 19998338 TI - Novel soluble HLA-A2/MELAN-A complexes selectively stain a differentiation defective subpopulation of CD8+ T cells in patients with melanoma. AB - Multimeric MHC I-peptide complexes containing phycoerythrin-streptavidin are widely used to detect and investigate antigen-specific CD8+ (and CD4+) T cells. Because such reagents are heterogeneous, we compared their binding characteristics with those of monodisperse dimeric, tetrameric and octameric complexes containing linkers of variable length and flexibility on Melan-A specific CD8+ T cell clones and peripheral blood mononuclear cells (PBMC) from HLA-A*0201(+) melanoma patients. Striking binding differences were observed for different defined A2/Melan-A(26-35) complexes on T cells depending on their differentiation stage. In particular, short dimeric but not octameric A2/Melan A(26-35) complexes selectively and avidly stained incompletely differentiated effector-memory T cells clones and populations expressing CD27 and CD28 and low levels of cytolytic mediators (granzymes and perforin). This subpopulation was found in PBMC from all six melanoma patients analyzed and proliferated on peptide stimulation with only modest phenotypic changes. By contrast influenza matrix(58 66) -specific CD8+ PBMC from nine HLA-A*0201(+) healthy donors were efficiently stained by A2/Flu matrix(58-61) multimers, but not dimer and upon peptide stimulation proliferated and differentiated from memory into effector T cells. Thus PBMC from melanoma patients contain a differentiation defective sub population of Melan-A-specific CD8+ T cells that can be selectively and efficiently stained by short dimeric A2/Melan- A(26-35) complexes, which makes them directly accessible for longitudinal monitoring and further investigation. PMID- 19998340 TI - Molecular analysis of the polymorphisms of thymidylate synthase on cell-free circulating DNA in blood of patients with advanced colorectal carcinoma. AB - As a key enzyme in folate metabolism, the thymidylate synthase (TS) is important for the synthesis of nucleotides. Its polymorphisms may affect the TS gene expression and the susceptibility for Fluoropyrimidine (FU)-based chemotherapies. In this study, we assessed the relationship between the TS genotypes and clinical outcome to 5-FU-based chemotherapy, and examined whether cell-free circulating DNA is applicable for these molecular analyses. We combined the variable number tandem repeat (VNTR) and single nucleotide (SNP) polymorphisms of the TS promoter and the deletion variants (1494del6) in the 3'UTR with the occurrence of loss of heterozygosity (LOH) at the microsatellite markers D18S59, D18S1140, and D18S976 mapped up- and downstream to the TS locus. Cell-free blood DNA, tumor tissues, and leukocytes of 51 patients with advanced colorectal cancer were used. Genotyping revealed linkage disequilibrium between TS promoter and 3'UTR (p = 0.03) in blood and leukocytes. Inverse associations of the response to therapy with the number of polymorphisms (p = 0.05) and the 494del6 polymorphism (p < 0.02) were detected on serum DNA. The quantification of serum DNA showed significant correlations with the 1494del6 variant (p = 0.006) in tumor tissues and the combined polymorphisms in leukocytes (p < 0.04). In contrast to the low LOH frequency in blood, LOH spanned more than one marker in the primary tumor of the majority of the patients suggesting the loss of the entire TS locus. Our data provide insight into the molecular diversity of the regulation of the TS gene expression. This is the first study that compares multi-variant TS genotypes in different clinical specimens. PMID- 19998339 TI - ERK/MAPK activation involves hypoxia-induced MGr1-Ag/37LRP expression and contributes to apoptosis resistance in gastric cancer. AB - We previously demonstrated that hypoxia increased the hypoxia-inducible factor (HIF-1)-dependent MGr1-Ag/37LRP expression, which enhanced adhesion of gastric cancer cells to laminin, inhibited drug-induced apoptosis and caused cell adhesion-mediated drug resistance (CAM-DR). Here, we investigated the role of extracellular-regulated kinase (ERK) 1/2 in the signaling mechanisms underlying these events. We found that hypoxia activated ERK activity in vitro and in vivo. Overexpression of mitogen-activated protein kinase (MAPK) kinase (MEK), which preferentially activated ERK, mimics, in a nonadditive way, hypoxia-induced activity of MGr1-Ag/37LRP promoter and expression of MGr1-Ag/37LRP. Furthermore, U0126, the MEK inhibitor, inhibited hypoxia- and MEK-induced MGr1-Ag/37LRP promoter activity in a dose-dependent manner. MEK inhibition also reversed hypoxia- and MEK-induced HIF-1 protein and its activity in a dose-dependent manner. We also investigated reactive oxygen species signaling this response. Exogenous addition of H(2)O(2) was sufficient to activate ERK in a dose-dependent profile. Reactive oxygen species scavengers of H(2)O(2) significantly inhibited hypoxia-induced ERK or HIF-1 activation and sequential expression of MGr1 Ag/37LRP. We also investigated the signaling in hypoxia-induced cell adhesion and apoptosis induced by vincristine. Hypoxia significantly enhanced adhesion of SGC7901 cells to laminin in a time-dependent manner, which might be inhibited by the MEK inhibitor U0126 and MGr1-Ag/37LRP siRNA. Consistent with results of adhesion assay, hypoxia-resistant apoptosis might be reversed by U0126 in a dose dependent manner. Our results suggest that hypoxia-elicited MGr1-Ag/37LRP expression activated by HIF-1 depends on ERK activation. These events are dependent of reactive oxygen intermediates. PMID- 19998342 TI - Early and late effects of the immunosuppressants rapamycin and mycophenolate mofetil on UV carcinogenesis. AB - Increased skin cancer risk in organ transplant recipients has been experimentally emulated with enhanced UV carcinogenesis from administering conventional immunosuppressants. However, newer generation immunosuppressive drugs, rapamycin (Rapa) and mycophenolate mofetil (MMF), have been shown to impair angiogenesis and outgrowth of tumor implants. To ascertain the overall effect on UV carcinogenesis, Rapa and MMF were admixed into the food pellets of hairless SKH1 mice receiving daily sub-sunburn UV dosages. With immunosuppressive blood levels neither of the drugs affected onset of tumors (<2 mm), but in contrast to MMF, Rapa significantly increased latency of large tumors (>or=4 mm, medians of 190 vs 125 days) and reduced their multiplicity (1.6 vs 4.5 tumors per mouse at 200 days). Interestingly, tumors (>2 mm) from the Rapa-fed group showed a reduction in UV-signature p53 mutations (39% vs 90%) in favor of mutations from putative base oxidation. This shift in mutation spectrum was not essentially linked to the reduction in large tumors because it was absent in large tumors similarly reduced in number when feeding Rapa in combination with MMF, possibly owing to an antioxidant effect of MMF. Significantly fewer tumor cells were Vegf-positive in the Rapa-fed groups, but a correspondingly reduced expression of Hif1alpha target genes (Vegf, Ldha, Glut1, Pdk1) that would indicate altered glucose metabolism with increased oxidative stress was not found. Remarkably, we observed no effect of the immunosuppressants on UV-induced tumor onset, and with impaired tumor outgrowth Rapa could therefore strongly reduce skin carcinoma morbidity and mortality rates in organ transplant recipients. PMID- 19998343 TI - Radiofrequency devices and hemolymphatic cancer. PMID- 19998344 TI - Hereditary hemorrhagic telangiectasia/avastin. AB - This is the first scientific report of hereditary hemorrhagic telangiectasia (HHT) epistaxis treatment by intranasal spraying of the vascular endothelial growth factor (VEGF) inhibitor bevacizumab (Avastin). Epistaxis in patients with HHT is a morbid, mortal condition that is difficult and unpleasant to manage. Nasal telangiectasia growth is modulated by VEGF, which is elevated in HHT patients. Bevacizumab is a VEGF inhibitor that diminishes epistaxis when administered intravenously or injected locally, or as reported here when sprayed topically onto the nasal mucosa. PMID- 19998345 TI - Calcium hydroxylapatite injection laryngoplasty for the treatment of presbylaryngis: long-term results. AB - OBJECTIVES/HYPOTHESIS: Presbylaryngis is a normal part of the aging process, but many people visit hospitals with communication difficulties. The authors evaluated the efficacy of calcium hydroxylapatite (CaHA) injection laryngoplasty in patients with presbylaryngis. STUDY DESIGN: Retrospective review. METHODS: Thirty-three patients with diagnosed presbylaryngis were administered a CaHA injection, and 17 of these patients without other vocal pathologies were included in the analysis. All 17 were male (mean age 65.9 years), mean follow-up duration was 335 days, and all injections were performed through the cricothyroid membrane under local anesthesia in a clinic. RESULTS: Subjective ratings, perceptual ratings, maximum phonation time, and closed quotients significantly improved after injection, and these improvements persisted without significant change for over 12 months. No major complications were encountered except for transient hematoma, pain, and a foreign body sensation. CONCLUSIONS: The authors conclude that CaHA injection laryngoplasty offers an effective and safe means of treating presbylaryngis. PMID- 19998346 TI - Life expectancy of screen-detected invasive breast cancer patients compared with women invited to the Nijmegen screening program. AB - BACKGROUND: Screening can lead to earlier detection of breast cancer and thus to an improvement in survival. The authors studied the life expectancy of women with screen-detected invasive breast cancer (patients) compared with women invited to the breast cancer screening program in Nijmegen, the Netherlands (comparison group). METHODS: Each patient diagnosed between 1975 and 2006 was randomly age matched with a woman invited in the same calendar year and free from breast cancer at the time of diagnosis of the patient. Survival analyses were performed to study differences in life expectancy. RESULTS: The life expectancy for 858 patients was 6 years shorter than for the comparison group. However, for 360 patients with small (<15 mm) invasive breast cancer, life expectancy was similar to that of the comparison group. In contrast, for patients detected with larger tumors (> or = 15 mm) the life expectancy was 6 to 12 years shorter, depending on tumor size. Furthermore, life expectancy was modified by screening history. For patients who had a negative screening examination 2 years before the detection of their breast cancer, the difference in life expectancy from the comparison group became smaller for the larger tumor sizes (> or = 15 mm). CONCLUSIONS: In conclusion, about 40% (360 of 858) of all women with invasive screen-detected breast cancer have the same life expectancy as women from the comparison group (reflecting the general population). For women diagnosed with larger tumors at diagnosis, life expectancy diminishes with increasing tumor size and is modified by screening history. PMID- 19998347 TI - Head and neck sequelae of torture. AB - OBJECTIVES/HYPOTHESIS: To increase awareness of torture among otolaryngologists, and to describe methods and complications of head and neck torture. STUDY DESIGN: Retrospective review. METHODS: Five cases of survivors of torture were evaluated in an otolaryngology practice in an urban hospital setting. RESULTS: The subjects presented with widely variable symptoms and physical manifestations related to the head and neck as a result of torture, in addition to psychiatric disease. Documentation of head and neck findings were essential to the asylum claim. CONCLUSIONS: Otolaryngologists serving immigrant and refugee populations must be familiar with methods and manifestations of torture involving the head and neck. PMID- 19998349 TI - Septal button insertion--the screw technique. PMID- 19998348 TI - Histologic evaluation of metastases in renal cell carcinoma with sarcomatoid transformation and its implications for systemic therapy. AB - BACKGROUND: Sarcomatoid features in renal cell carcinoma may represent an aggressive subclone arising from the primary tumor. The patterns of metastases for these tumors were evaluated to determine if sarcomatoid features were retained at metastasis and whether the percentage of sarcomatoid features in the primary tumor influenced spread. METHODS: All patients with sarcomatoid features found at nephrectomy with synchronous or metachronous resection of metastases were evaluated. The histology, grade, and percentage of sarcomatoid features in the primary and metastatic site were recorded. The association between percentage of sarcomatoid features, grade, histology, and pattern of metastases was evaluated. RESULTS: Thirty-two patients were identified with sarcomatoid features and resected metastases. Fifty-two metastatic sites were evaluated. A single histologic appearance (sarcomatoid or carcinomatoid) was present in 50 of 52 sites (96%). Thirty sites (58%) demonstrated only a sarcomatoid pattern, whereas 20 (38%) contained only a carcinoma pattern. Histology and carcinoma grade did not influence metastatic pattern; however, greater percentage of sarcomatoid features was associated with the presence of distant sarcomatoid histology. A cutoff of 30% sarcomatoid features in the primary tumor was useful in predicting systemic sarcomatoid histology. CONCLUSIONS: Sarcomatoid elements are frequently observed in the metastases of primary tumors with sarcomatoid features, and these metastases generally contain a solitary pattern supporting the subclone hypothesis. However, both components can metastasize in the same patient. The percentage of sarcomatoid features influences the pattern of spread, and patients with >30% sarcomatoid features in the primary tumor frequently have distant sarcomatoid histology. This cutpoint may be helpful for inclusion criteria for future clinical trials. PMID- 19998350 TI - Are patients of low socioeconomic status receiving suboptimal management for pancreatic adenocarcinoma? AB - BACKGROUND: The objective of this study was to define the effects of socioeconomic status (SES) and other demographic variables on outcomes for patients with pancreatic adenocarcinoma. METHODS: Florida cancer registry and inpatient hospital data were queried for pancreatic adenocarcinoma diagnosed from 1998 to 2002. RESULTS: In total, 16,104 patients were identified. Low SES (LSES) patients were younger at diagnosis (P < .001) but presented with similar disease stage and tumor grade. LSES patients were less likely to receive surgical extirpation (16.5% vs 19.8%; P < .001), chemotherapy (30.7% vs 36.4%; P < .001), or radiotherapy (14.3% vs 16.9%; P = .003). Among surgical patients, 30-day mortality was significantly higher (5.1% vs 3.7%; P < .001) and overall median survival was significantly worse (5.0 months vs 6.2 months; P < .001) in the LSES cohorts. Although surgical patients who were treated at teaching facilities (TF) did significantly better; an increased 30-day surgical mortality (2.2% vs 1.3%; P < .001) and decreased median survival (5 months for poverty level >15% vs 6.2 months for poverty level <5%; P < .001) also were observed for patients of LSES. In a multivariate analysis that corrected for patient comorbidities, significant independent predictors of a poorer prognosis included LSES (hazard ratio [HR], 1.09); treatment at a non-TF (HR, 1.09); and failure to receive surgical extirpation (HR, 1.92), chemotherapy (HR 1.41), or radiation (HR 1.25). CONCLUSIONS: Patients of LSES were less likely to receive surgical extirpation, chemotherapy, or radiation and had significantly higher perioperative and long term mortality rates. A greater understanding of the barriers to providing optimal care and identifying means for improving successful delivery of therapies to the poor with pancreatic cancer are needed. PMID- 19998351 TI - Long-term outcomes after hepatic resection for colorectal metastases in young patients. AB - BACKGROUND: Long-term outcomes after hepatectomy for colorectal liver metastases in relatively young patients are still unknown. The aim of the current study was to evaluate long-term outcomes in patients < or = 40 years old, and to compare them with patients >40 years old. METHODS: All consecutive patients who underwent hepatectomy for colorectal liver metastases at the authors' hospital between 1990 and 2006 were included in the study. Patients < or = 40 years old were compared with all other patients treated during the same period. Overall survival (OS), progression-free survival (PFS), and disease-free survival (DFS) rates were determined, and prognostic factors were identified. RESULTS: In total, 806 patients underwent hepatectomy for colorectal liver metastases, of whom 56 (7%) were aged < or = 40 years. Among the young patients, more colorectal liver metastases were present at diagnosis, and they were more often diagnosed synchronous with the primary tumor. Five-year OS was 33% in young patients, compared with 51% in older patients (P = .12). Five-year PFS was 2% in young patients, compared with 16% in older patients (P < .001). DFS rates were comparable between the groups (17% vs 23%, P = .10). At multivariate analysis, age < or = 40 years was identified as an independent predictor of poor PFS. CONCLUSIONS: In young patients, colorectal liver metastases seem to be more aggressive, with a trend toward lower OS, more disease recurrences, and a significantly shorter PFS after hepatectomy. However, DFS rates were comparable between young and older patients, owing to an aggressive multimodality treatment approach, consisting of chemotherapy and repeat surgery. Therefore, physicians should recognize the poor outcome of colorectal liver metastases in young patients and should consider an aggressive approach to diagnosis and early treatment. PMID- 19998352 TI - Effects of venue-specific state clean indoor air laws on smoking-related outcomes. AB - A large literature has documented relationships between state clean indoor air laws (SCIALs) and smoking-related outcomes in the United States. These laws vary within states over time and across venues such as schools, government buildings, and bars. Few studies, however, have evaluated whether the effects of SCIALs are plausibly concentrated among workers who should have been directly affected because they worked at locations covered by the venue-specific restrictions. We fill this gap in the literature using data on private sector workers, government employees, school workers, eating and drinking place workers, and bartenders from the 1992-2007 Tobacco Use Supplements to the Current Population Survey. Our quasi experimental models indicate robust effects of SCIALs restricting smoking in bars: these laws significantly increased the presence of workplace smoking restrictions as reported by bartenders and reduced the fraction of bartenders who smoke. We do not, however, find that SCIALs in private workplaces, government workplaces, schools, or restaurants increased the presence of workplace smoking restrictions among groups of workers working in venues covered by these laws. This suggests that the smoking reductions associated with SCIALs in previous research are unlikely to have been directly caused by effects of workplace smoking restrictions on workers. PMID- 19998354 TI - Adaptation of object descriptions to a partner under increasing communicative demands: a comparison of children with and without autism. AB - This study compared the object descriptions of school-age children with high functioning autism (HFA) with those of a matched group of typically developing children. Descriptions were elicited in a referential communication task where shared information was manipulated, and in a guessing game where clues had to be provided about the identity of an object that was hidden from the addressee. Across these tasks, increasingly complex levels of audience design were assessed: (1) the ability to give adequate descriptions from one's own perspective, (2) the ability to adjust descriptions to an addressee's perspective when this differs from one's own, and (3) the ability to provide indirect yet identifying descriptions in a situation where explicit labeling is inappropriate. Results showed that there were group differences in all three cases, with the HFA group giving less efficient descriptions with respect to the relevant context than the comparison group. More revealing was the identification of distinct adaptation profiles among the HFA participants: those who had difficulty with all three levels, those who displayed Level 1 audience design but poor Level 2 and Level 3 design, and those demonstrated all three levels of audience design, like the majority of the comparison group. Higher structural language ability, rather than symptom severity or social skills, differentiated those HFA participants with typical adaptation profiles from those who displayed deficient audience design, consistent with previous reports of language use in autism. PMID- 19998355 TI - Investigating the disorder-order transition of calmodulin binding domain upon binding calmodulin using molecular dynamics simulation. AB - We have studied the conformational transition of the calmodulin binding domains (CBD) in several calmodulin-binding kinases, in which CBD changes from the disordered state to the ordered state when binding with calmodulin (CaM). Targeted molecular dynamics simulation was used to investigate the binding process of CaM and CBD of CaM-dependent kinase I (CaMKI-CBD). The results show that CaMKI-CBD began to form an alpha-helix and the interaction free energy between CaM and CaMKI-CBD increased once CaM fully encompassed CaMKI-CBD. Two series of CaM/CBD complex systems, including the complexes of CaM with the initially disordered and the final ordered CBD, were constructed to study the interaction using molecular dynamics simulations. Our analyses suggest that the VDW interaction plays a dominant role in CaM/CBD binding and is a key factor in the disorder-order transition of CBD. Additionally, the entropy effect is not in favor of the formation of the CaM/CBD complex, which is consistent with the experimental evidence. Based on the results, it appears that the CBD conformational change from a non-compact extended structure to compact alpha helix is critical in gaining a favorable VDW interaction and interaction free energy. PMID- 19998356 TI - Attention deficit/hyperactivity disorder symptoms moderate cognition and behavior in children with autism spectrum disorders. AB - Recent estimates suggest that 31% of children with autism spectrum disorders (ASD) meet diagnostic criteria for attention deficit/hyperactivity disorder (ADHD), and another 24% of children with ASD exhibit subthreshold clinical ADHD symptoms. Presence of ADHD symptoms in the context of ASD could have a variety of effects on cognition, autistic traits, and adaptive/maladaptive behaviors including: exacerbating core ASD impairments; adding unique impairments specific to ADHD; producing new problems unreported in ASD or ADHD; having no clear impact; or producing some combination of these scenarios. Children with ASD and co-morbid ADHD symptoms (ASD+ADHD; n = 21), children with ASD without ADHD (ASD; n = 28), and a typically developing control group (n = 21) were included in the study; all groups were matched on age, gender-ratio, IQ, and socioeconomic status. Data were collected on verbal and spatial working memory, response inhibition, global executive control (EC), autistic traits, adaptive functioning, and maladaptive behavior problems. In this sample, the presence of ADHD symptoms in ASD exacerbated impairments in EC and adaptive behavior and resulted in higher autistic trait, and externalizing behavior ratings. ADHD symptoms were also associated with greater impairments on a lab measure of verbal working memory. These findings suggest that children with ASD+ADHD symptoms present with exacerbated impairments in some but not all domains of functioning relative to children with ASD, most notably in adaptive behavior and working memory. Therefore, ADHD may moderate the expression of components of the ASD cognitive and behavioral phenotype, but ASD+ADHD may not represent an etiologically distinct phenotype from ASD alone. PMID- 19998358 TI - Mechanical peeling of free-standing single-walled carbon-nanotube bundles. AB - An in situ electron microscopy study is presented of adhesion interactions between single-walled carbon nanotubes (SWNTs) by mechanically peeling thin free standing SWNT bundles using in situ nanomanipulation techniques inside a high resolution scanning electron microscope. The in situ measurements clearly reveal the process of delaminating one SWNT bundle from its originally bound SWNT bundle in a controlled-displacement manner and capture the deformation curvature of the delaminated SWNT bundle during the peeling process. A theoretical model based on nonlinear elastica theory is employed to interpret the measured deformation curvatures of the SWNTs and to quantitatively evaluate the peeling force and the adhesion strength between bundled SWNTs. The estimated adhesion energy per unit length for each pair of neighboring tubes in the peeling interface based on our peeling experiments agrees reasonably well with the theoretical value. This in situ peeling technique provides a potential new method for separating bundled SWNTs without compromising their material properties. The combined peeling experiments and modeling presented in this paper will be very useful to the study of the adhesion interactions between SWNTs and their nonlinear mechanical behaviors in the large-displacement regime. PMID- 19998357 TI - Preparation and unique pH-responsive properties of novel biodegradable nanocapsules composed of poly(gamma-glutamic acid) and chitosan as weak polyelectrolytes. AB - Novel biodegradable hollow nanocapsules composed of two kinds of weak polyelectrolytes, CT and gamma-PGA, were successfully prepared by the deposition of their LbL-assembled films onto silica particles and the subsequent removal of the silica. These CT-gamma-PGA hollow nanocapsules showed unique size increases at pH = 1.0, due to the swelling of capsule membranes induced by electrostatic repulsions between ammonium groups of CT components. On the other hand, no significant changes in the capsule size were observed at pH = 4.0, 7.0, and 10. By using the CT-gamma-PGA nanocapsules, the release of encapsulated substances in response to acidic pH values was accomplished. PMID- 19998359 TI - Recent advances in the analysis of antibiotics by CE and CEC. AB - This article reviews the latest advances in the development of analytical methodologies by CE and CEC for the analysis of different groups of antibiotics. CZE and MEKC have been the most employed CE modes. Analytical information on experimental conditions, detection mode, and LOD are provided to the readers. In addition, the main applications performed in the pharmaceutical, biological, food, and environmental fields have been reviewed emphasizing the sample preparation requirements. The publications reviewed cover the period of time from June 2007 to May 2009. PMID- 19998360 TI - Assessing systemic drug exposure in repeated dose toxicity studies in the case of complete and incomplete sampling. AB - Repeated dose toxicity studies are performed to characterize the toxicological profile of a test compound following repeated administrations. The findings and interpretations from these systemic exposure studies in animals are essential for designing subsequent studies and evaluating the safety of the test item for humans. Blood samples for assessment of systemic exposure are usually collected on day one and at the end of the study with multiple dosings of the compound in between. Restrictions in blood volume often require an incomplete sampling design, in which each animal contributes sample measurements at some but not all time points. In this manuscript we derive an estimator for the ratio of area under the concentration versus time curves (AUCs), a frequently used measure of exposure to a compound, and a corresponding confidence interval to assess differences in exposure as well as equivalence between first and repeated administration that is applicable in such sparse sampling designs as well as complete data situations. An illustrative example is provided and the statistical properties of the proposed estimator, which incorporates the dependencies of measurements between first and repeated dosings as well as the dependency inherent in repeated sampling for each dosing, is studied asymptotically as well as in simulation. PMID- 19998361 TI - Embedded phases: a way to active and stable catalysts. AB - Industrial catalysts are typically made of nanosized metal particles, carried by a solid support. The extremely small size of the particles maximizes the surface area exposed to the reactant, leading to higher reactivity. Moreover, the higher the number of metal atoms in contact with the support, the better the catalyst performance. In addition, peculiar properties have been observed for some metal/metal oxide particles of critical sizes. However, thermal stability of these nanostructures is limited by their size; smaller the particle size, the lower the thermal stability. The ability to fabricate and control the structure of nanoparticles allows to influence the resulting properties and, ultimately, to design stable catalysts with the desired characteristics. Tuning particle sizes provides the possibility to modulate the catalytic activity. Unique and unexpected properties have been observed by confining/embedding metal nanoparticles in inorganic channels or cavities, which indeed offers new opportunities for the design of advanced catalytic systems. Innovation in catalyst design is a powerful tool in realizing the goals of more green, efficient and sustainable industrial processes. The present Review focuses on the catalytic performance of noble metal- and non precious metal-based embedded catalysts with respect to traditional impregnated systems. Emphasis is dedicated to the improved thermal stability of these nanostructures compared to conventional systems. PMID- 19998362 TI - Transition metal-tungsten bimetallic catalysts for the conversion of cellulose into ethylene glycol. PMID- 19998363 TI - C factors pinpoint resource utilization in chemical industrial processes. AB - This Full Paper illustrates the use of the C factor (CO(2)/product mass ratio) as a parameter to evaluate the CO(2)-burden of a product. The C factor contains information of the total amount of CO(2) emitted in order to produce a product, and thus enables a direct comparison of different processes from a CO(2) aspect. We illustrate how this simple concept can be used to evaluate different resource types and processes. The C factors for different chemicals such as methanol, synfuels, and acetic acid are calculated for oil, coal, natural gas, and biomass. Based on these calculations, the combination of biomass and natural gas is an attractive alternative to coal, leading to products that have significantly lower C factors. PMID- 19998364 TI - Brain-behavior relationships in young traumatic brain injury patients: DTI metrics are highly correlated with postural control. AB - Traumatic brain injury (TBI) is a major cause of impairment and functional disability in children and adolescents, including deterioration in fine as well as gross motor skills. The aim of this study was to assess deficits in sensory organization and postural ability in a young group of TBI patients versus controls by using quantitative force-platform recordings, and to test whether balance deficits are related to variation in structural properties of the motor and sensory white matter pathways. Twelve patients with TBI and 14 controls (aged 8-20 years) performed the Sensory Organisation Test (SOT) protocol of the EquiTest (Neurocom). All participants were scanned using Diffusion Tensor Imaging (DTI) along with standard anatomical scans. Quantitative comparisons of DTI parameters (fractional anisotropy, axial and radial diffusivity) between TBI patients and controls were performed. Correlations between DTI parameters and SOT balance scores were determined. Findings revealed that the TBI group scored generally lower than the control group on the SOT, indicative of deficits in postural control. In the TBI group, reductions in fractional anisotropy were noted in the cerebellum, posterior thalamic radiation, and corticospinal tract. Degree of white matter deterioration was highly correlated with balance deficits. This study supports the view that DTI is a valuable tool for assessing the integrity of white matter structures and for selectively predicting functional motor deficits in TBI patients. PMID- 19998365 TI - Identifying spatially overlapping local cortical networks with MEG. AB - Recent modelling studies (Hadjipapas et al. [2009]: Neuroimage 44:1290-1303) have shown that it may be possible to distinguish between different neuronal populations on the basis of their macroscopically measured (EEG/MEG) mean field. We set out to test whether the different orientation columns contributing to a signal at a specific cortical location could be identified based on the measured MEG signal. We used 1.5deg square, static, obliquely oriented grating stimuli to generate sustained gamma oscillations in a focal region of primary visual cortex. We then used multivariate classifier methods to predict the orientation (left or right oblique) of the stimuli based purely on the time-series data from this one location. Both the single trial evoked response (0-300 ms) and induced post transient power spectra (300-2,300 ms, 20-70 Hz band) due to the different stimuli were classifiable significantly above chance in 11/12 and 10/12 datasets respectively. Interestingly, stimulus-specific information is preserved in the sustained part of the gamma oscillation, long after perception has occurred and all neuronal transients have decayed. Importantly, the classification of this induced oscillation was still possible even when the power spectra were rank transformed showing that the different underlying networks give rise to different characteristic temporal signatures. PMID- 19998366 TI - Structural brain differences and cognitive functioning related to body mass index in older females. AB - Little is known about the effect of obesity on brain structures and cognition in healthy older adults. This study examined the association between body mass index (BMI), regional volume differences in gray and white matter measured by magnetic resonance imaging (MRI), and cognitive functioning in older females. Participants included 95 community-dwelling older females (ages 52-92 years) who underwent extensive neuropsychological testing and high-resolution MRI scanning. Optimized voxel-based morphometry techniques were employed to determine the correlation between BMI and regional gray and white matter volumes. Volumes of significant regions were then correlated with cognitive functioning. Higher BMI was associated with decreased gray matter volumes in the left orbitofrontal, right inferior frontal, and right precentral gyri, a right posterior region including the parahippocampal, fusiform, and lingual gyri, and right cerebellar regions, as well as increased volumes of white matter in the frontal, temporal, and parietal lobes, even when hypertension was considered. Compared to normal weight women, obese women performed poorer on tests of executive functioning. Smaller gray matter volume in the left orbitofrontal region was associated with lower executive functioning. Additionally, despite the lack of significant group differences in memory and visuomotor speed, gray and white matter volumes predicted performance on these measures. The results provide additional evidence for a negative link between increased body fat and brain functioning in older females. PMID- 19998367 TI - 3D mapping of brain differences in native signing congenitally and prelingually deaf subjects. AB - In the prelingual and congenital deaf, functional reorganization is known to occur throughout brain regions normally associated with hearing. However, the anatomical correlates of these changes are not yet well understood. Here, we perform the first tensor-based morphometric analysis of voxel-wise volumetric differences in native signing prelingual and congenitally deaf subjects when compared with hearing controls. We obtained T1-weighted scans for 14 native signing prelingual and congenitally deaf subjects and 16 age- and gender-matched controls. We used linear and fluid registration to align each image to a common template. Using the voxel-wise determinant of the Jacobian of the fluid deformation, significant volume increases, of up to 20%, were found in frontal lobe white matter regions including Broca's area, and adjacent regions involved in motor control and language production. A similar analysis was performed on hand-traced corpora callosa. A strong trend for group differences was found in the area of the splenium considered to carry fibers connecting the temporal (and occipital) lobes. These anatomical differences may reflect experience-mediated developmental differences in myelination and cortical maturation associated with prolonged monomodal sensory deprivation. PMID- 19998368 TI - HNF1B and JAZF1 genes, diabetes, and prostate cancer risk. AB - BACKGROUND: Epidemiologic studies have shown that men with type II diabetes have a lower risk of prostate cancer than non-diabetic men. Recently, common variants in two genes, HNF1B and JAZF1, were found to be associated with both of these diseases. METHODS: We examined whether the relationship between HNF1B and JAZF1 variants and decreased prostate cancer risk may potentially be mediated through diabetes in two large prospective studies, the Cancer Prevention Study II Nutrition Cohort and the Prostate, Lung, Colorectal, and Ovarian Cancer Screening Trial. RESULTS: Three HNF1B SNPS, rs11649743, rs4430796, and rs7501939, were associated with decreased risk of prostate cancer and were also associated, with marginal statistical significance, with increased risk of diabetes. The JAZF1 SNPs rs6968704 and rs10486567 were associated with decreased risk of prostate cancer but were not associated with diabetes. All five SNP-prostate cancer relationships did not substantially differ when the analyses were stratified by diabetic status or when diabetic status was controlled for in the model. Furthermore, the association of diabetes with prostate cancer was not altered when the SNPs were included in the logistic model. CONCLUSIONS: These findings indicate that the HNF1B variants are directly associated with both diabetes and prostate cancer, that diabetes does not mediate these gene variant-prostate cancer relationships, and the relationship between these diseases is not mediated through these gene variants. PMID- 19998369 TI - The impact of HMG-CoA reductase therapy on serum PSA. AB - BACKGROUND: 3-hydroxy-3-methyl-glutaryl-CoA (HMG-CoA) reductase inhibitors, otherwise known as statins, inhibit the enzyme that controls the conversion of HMG-CoA to mevalonate, a precursor for cholesterol. Statins may be important to prostate cancer biology by inhibiting cell growth, inflammation, and oxidative stress. The purpose of this study was to assess the influence of statin therapy on serum prostate-specific antigen (PSA) levels. METHODS: The computerized medical records at the University of Rochester Medical Center were used to identify men who filled statin prescriptions between May 31st, 2008 and September 30th, 2008. Men with at least one PSA assay performed within 2 years before and at least one PSA assay performed within 1 year after starting a statin medication were included. The primary endpoint was the change in PSA concentration computed as the difference between PSA levels before and after starting a statin medication. Paired t-tests were used to analyze the mean differences in PSA values. RESULTS: A total of 962 patients were identified. The mean difference in serum PSA level after statin administration was -0.29 ng/ml (-8.04%). Subgroup analyses for mean PSA concentration change before and after statin administration by age group revealed: 50-59 years old (-0.1609, 95% CI: -0.2444, -0.0775, P < 0.0002), 60-69 years old (-0.3393, 95% CI: -0.4641, -0.2145, P < 0.0001), and >70 years old (-0.351, 95% CI: -0.490, -0.212, P < 0.0001). CONCLUSIONS: These observations suggest a statistically significant reduction in serum PSA level that is associated with the onset of statin therapy. PMID- 19998370 TI - Preventable drug related mortality in a Swedish population. AB - PURPOSE: Several studies indicate that the medical burden of fatal adverse drug reactions (FADRs) is significant, but the preventability of FADRs in the general population is largely unknown. The aim of this study was to determine the proportion of preventable FADRs and preventable fatal drug poisonings (FDPs) in a Swedish population. METHODS: Previously, a population-based sample of 1574 deceased subjects was scrutinised for FADRs and FDPs using relevant case records, including death certificates, medical charts and medico-legal files. Forty-nine cases (3%) of FADRs and nine cases (0.6%) of FDPs were identified in 57 subjects. In this study, the preventability of all these identified FADRs and FDPs was evaluated by clinical experts in a stepwise manner, applying a set of predefined and well established preventability criteria. Only cases for which consensus was achieved were included in the study. RESULTS: Of 49 FADRs, 14% (seven fatalities) was considered definitely or possibly preventable and four of these were due to the presence of a contraindication for the drug. All nine FDPs were considered possibly preventable. As one subject had a combination of an FADR and an FDP, a total of 15 persons (26%) were considered having a definitely or possibly preventable FADR or FDP, corresponding to 0.95% of all deceased subjects in Sweden. CONCLUSIONS: The results of this study indicate that approximately one fourth of FADRs and FDPs could be prevented. Therefore, an increased awareness of the possibility to reduce the risk of fatal events due to pharmaceutical drugs is warranted. PMID- 19998371 TI - Law, ethics, religion, and clinical translation in the 21st century--a discussion with John Sinden. PMID- 19998372 TI - Law, ethics, religion, and clinical translation in the 21st century. PMID- 19998374 TI - Screening of closantel and rafoxanide in animal muscles by HPLC with fluorescence detection and confirmation using MS. AB - A sensitive and selective LC-FLD method was described for determination of closantel and rafoxanide in bovine and ovine muscles. Bovine and ovine muscles were extracted with ACN and acetone mixture (80:20, v/v). After cleanup with Oasis MAX SPE cartridges, the sample was analyzed by LC-FLD using the control point approach. No false-negative result was observed at or below maximum residue limits and the false-positive rate was below 5%. Suspected positive sample was confirmed by LC-MS/MS. This method was suitable for screening of large batch of samples and hence considerably reduced the time and cost required for quantitation and confirmatory analyses. PMID- 19998373 TI - Identification of vitronectin as an extrinsic inducer of cancer stem cell differentiation and tumor formation. AB - There is mounting evidence that tumors are initiated by a rare subset of cells called cancer stem cells (CSCs). CSCs are generally quiescent, self-renew, form tumors at low numbers, and give rise to the heterogeneous cell types found within a tumor. CSCs isolated from multiple tumor types differentiate both in vivo and in vitro when cultured in serum, yet the factors responsible for their differentiation have not yet been identified. Here we show that vitronectin is the component of human serum driving stem cell differentiation through an integrin alpha V beta 3-dependent mechanism. CSCs cultured on vitronectin result in downregulation of stem cell genes, modulation of differentiation markers, and loss of beta-catenin nuclear localization. Blocking integrin alpha V beta 3 inhibits differentiation and subsequently tumor formation. Thus, CSCs must be engaged by one or more extracellular signals to differentiate and initiate tumor formation, defining a new axis for future novel therapies aimed at both the extrinsic and intracellular pathways. PMID- 19998375 TI - Application of bromoacetate-substituted beta-CD-bonded silica particles as chiral stationary phase for HPLC. AB - Bromoacetate-substituted [3-(2-O-beta-cyclodextrin)-2-hydroxypropoxy]propylsilyl appended silica particles (BACD-HPS), an important and useful synthetic intermediate for preparation of novel types of macrocycles-capped beta-CD-bonded silica particles including crown ether/cyclam/calix[4]arene-capped beta-CD-bonded silica particles, have been prepared and used as chiral stationary phase for HPLC. This synthetic stationary phase is characterized by means of elemental analysis. For the first time, the chromatographic behavior of BACD-HPS was systematically evaluated with several disubstituted benzenes and some chiral drug compounds under both normal and RP conditions in HPLC. The results show that BACD HPS has excellent selectivity for the separation of aromatic positional isomers and chiral isomers of some drug compounds when used as stationary phase in HPLC. PMID- 19998376 TI - Determination of melamine in royal jelly and lyophilized powder using ultrasonically assisted extraction by ion-pair HPLC with UV detection. AB - A rapid method for the analysis of melamine in royal jelly (RJ) and RJ lyophilized powder (RJLP) was developed using ion-pair RP-HPLC coupled with UV detector. The method utilized an optimized buffer system to avoid the elution of melamine near the column void volume and improve retention of melamine in a generic C8 chromatographic column. In addition, sample preparation included deproteination, ultrasonic-assisted extraction, and cleanup on a mixed-mode cation exchange extraction cartridge. The extraction procedure was optimized with regard to the amount of extraction solvent and the duration of sonication for RJ and RJLP samples. The following criteria were used to validate the HPLC-UV detection method: selectivity, linearity, precision, LOD, and LOQ. Correlation coefficient was higher than 0.999 by applying the linear regression model based on the least square method with a weighting factor (1/x). Precision was evaluated as repeatability and intermediary precision with RSD of less than 15%. The mean percentage recoveries of melamine were varied from 72.5 to 90.5% for RJ and RJLP. This approach will be of particular utility for the evaluation of melamine residue level and routine monitoring of melamine in RJ and RJLP samples. PMID- 19998377 TI - Highly selective GC stationary phases consisting of binary mixtures of polymeric ionic liquids. AB - GC stationary phases composed of binary mixtures of two polymeric ionic liquids (PILs), namely, poly(1-vinyl-3-hexylimidazolium) bis[(trifluoromethyl)sulfonyl]imide (poly(ViHIm-NTf(2)))/poly(1-vinyl-3 hexylimidazolium) chloride (poly(ViHIm-Cl)) and poly(1-vinyl-3 hexadecylimidazolium) bis[(trifluoromethyl)sulfonyl]imide (poly(ViHDIm NTf(2)))/poly(1-vinyl-3-hexadecylimidazolium) chloride (poly(ViHDIm-Cl)), were evaluated in terms of their on-set bleed temperature and separation selectivity. A total of six neat or binary PIL stationary phases were characterized using the solvation parameter model to investigate the effects of the polymeric cation and anion and PIL composition on the system constants of the resulting stationary phases. The hydrogen bond basicity of the mixed poly(ViHIm-NTf(2))/poly(ViHIm-Cl) stationary phases was enriched linearly with the increase in the poly(ViHIm-Cl) content. Results revealed that tuning the composition of the stationary phase allowed for fine control of the retention factors and separation selectivity for alcohols and carboxylic acids as well as selected ketones, aldehydes, and aromatic compounds. A reversal of elution order was observed for particular classes of analytes when the weight percentage of the chloride-based PIL was increased. PMID- 19998378 TI - Marker compounds for the determination of retention factors in EKC. AB - EKC and its sub-techniques, such as MEKC and microemulsion EKC, have attracted wide interest in recent years. Investigations on this topic have covered several analytical applications, but attention has also been paid more and more to basic studies. This review provides an overview of the different approaches to calculating retention factors, which express the ratio of the amount of sample component in the pseudostationary and mobile phases. Special attention is given to the selection of markers for the determination of the electrophoretic mobility or migration time of a marker describing the behavior of the pseudostationary phase in EKC. Introduction of a hydrophobic marker is by far the most common approach, but the use of a homologous series of compounds is also quite popular. In addition, other possible approaches found in the literature will be described. PMID- 19998379 TI - Determination of phthalates in fruit jellies by dispersive SPE coupled with HPLC MS. AB - In this study, a simple, rapid and sensitive method for the determination of five phthalates including dimethyl phthalate, diethyl phthalate, dipropyl phthalate, benzyl butyl phthalate, and dicyclohexyl phthalate in fruit jellies by LC coupled with MS has been developed. Samples were pretreated by a dispersive SPE method, termed QuEChERS, which is an acronym for quick, easy, cheap, effective, rugged, and safe. The standard calibration curves were linear for all the analytes over the concentration range of 10-250 ng/mL, and the correlation coefficients ranged from 0.9976 to 0.9991. The LODs and LOQs were in the ranges of 0.09-3.68 ng/mL and 0.28-11.25 ng/mL, respectively. The accuracy of this method was evaluated by measuring the recovery from spiked samples. The recoveries of all five phthalates from samples spiked at three different concentrations (0.01, 0.03, and 0.05 mg/kg), were in the ranges of 83.5-103.9%, 86.7-95.8%, and 87.1-95.2%, respectively. The RSD values for the samples spiked at 0.01, 0.03, and 0.05 mg/kg ranged from 2.0-7.6%, 1.4-6.4%, and 1.2-3.8%, respectively. The method has been used for the analysis of real samples and BBP and DEP were found in real samples. PMID- 19998380 TI - Multistep mechanism of probiotic bacterium, the effect on innate immune system. AB - The etiology and pathogenesis of inflammatory bowel disease are still not fully understood. However, evidence from both animal models and clinical observations suggests luminal bacteria as the most probable inducer of this disease. The intestinal bacterial microbiota may be modified by dietary addition of viable probiotic bacteria, thereby constituting an alternative approach to disease prevention and treatment. The aim of this study was to evaluate and compare the effects of two probiotic regiments; Lactobacillus GG and a mixture of Streptococcus thermophilus, Lactobacillus acidophilus, and Bifidobacterium lactis (YO-MIX Y 109 FRO 1000) in both normal and trinitrobenzenesulfonic acid colitis induced rats. Colon morphology and damage were evaluated histologically; colonic tissues were used for mRNA analysis, using real-time PCR. Administration of both probiotics reduced the expression of proinflammatory cytokines tumor necrosis factor-alpha and IL-6 and increased the expression of mucin 2 in compared with colitis group and reduced the inflammatory response. These results provide additional support for the positive effect of probiotics in the gut and may shed light on the mechanism by which probiotic bacteria exert their action in an animal model. PMID- 19998381 TI - Dietary intake and depressive symptoms: a systematic review of observational studies. AB - The importance of research into the possible role of dietary intake in depressive symptoms is emphasized by the fact that diet is modifiable. We systematically reviewed observational studies investigating the association between dietary intake and depressive symptoms published in English as of December 2008. Using the PubMed database, 34 publications (23 cross-sectional, 10 prospective cohort, and 1 case-control studies) were identified. The number of subjects (n=80 27,111), age of subjects (15-97 years), dietary assessment method (dietary record, diet history interview, and validated and non-validated dietary questionnaire), depressive symptom assessment (discharge diagnosis, established scale, and self-reported information) varied among studies. Dietary variables most frequently investigated included long chain n-3 PUFA, fish, folate, and other B vitamins. Most studies found no association between dietary variables and depressive symptoms. However, most studies included at least one important methodological limitation, such as no inference for causality, unreliable or rough assessment of diet or depressive symptoms, inadequate treatment of potential confounding factors, and ignorance of the possible mediating or confounding influence of other dietary variables. Further evidence from well designed observational studies is required to confirm or refute the association between dietary intake and depressive symptoms in free-living settings. PMID- 19998382 TI - Increased plasma n-3 polyunsaturated fatty acid is associated with improved insulin sensitivity in type 2 diabetes in China. AB - Increased tissue n-3 polyunsaturated fatty acid (PUFA) is associated with improved insulin sensitivity in type 2 diabetes. However, this relationship among Chinese is not clear. To investigate the relationship between plasma phospholipids (PL) fatty acid composition and insulin resistance (IR) in type 2 diabetes mellitus, 186 type 2 diabetes and 180 healthy subjects were studied in this case-control study. In the sex, age and BMI controlled partial correlation, homeostasis model assessment (HOMA)-IR and blood glucose was significantly negatively correlated with plasma PL n-3 PUFA, 20:5n-3 and ratio of n-3:n-6 (p<0.01), and positively correlated with n-6 PUFA (p<0.001) and saturated fatty acid (p<0.05) in the diabetes patients. PL 22:6n-3 was also significantly negatively correlated with HOMA-IR (p<0.01), but not with blood glucose. Fasting insulin was significantly negatively correlated with plasma PL n-3 PUFA, 20:5n-3, 22:6n-3 and ratio of n-3:n-6 (p<0.01). The 18:3n-3 was not associated with HOMA IR and fasting insulin. The results suggested that increased plasma PL n-3 PUFA, 20:5n-3, 22:6n-3 and ratio of n-3:n-6 PUFA was associated with decreased HOMA-IR in type 2 diabetes. Increased plasma PL n-3 PUFA improves insulin sensitivity in type 2 diabetes. PMID- 19998383 TI - Diosgenin attenuates inflammatory changes in the interaction between adipocytes and macrophages. AB - Obese adipose tissues are characterized by the enhanced infiltration of macrophages. It is considered that the paracrine loop involving monocyte chemoattractant protein-1, tumor necrosis factor-alpha, and the free fatty acid between adipocytes and macrophages establishes a vicious cycle that aggravates inflammatory changes and insulin resistance in obese adipose tissues. Diosgenin, a saponin aglycon found in a variety of plants, has anti-inflammatory properties. In the present study, we examined the effect of diosgenin on the inflammatory changes in the interaction between adipocytes and macrophages. A coculture of 3T3 L1 adipocytes and RAW 264 macrophages markedly enhanced the production of tumor necrosis factor-alpha, monocyte chemoattractant protein-1, and nitric oxide compared with the sum of their single cultures; however, treatment with diosgenin inhibited the production of these proinflammatory mediators. Diosgenin also suppressed the inflammation in RAW 264 macrophages that was induced by the conditioned medium derived from 3T3-L1 adipocytes. Furthermore, diosgenin inhibited the conditioned medium-induced degradation of inhibitor kappaB and the phosphorylation of c-jun N-terminal kinase in macrophages. These results indicate that diosgenin exhibits anti-inflammatory properties in the interaction of adipocytes and macrophages by inhibiting the inflammatory signals in macrophages. Diosgenin may be useful for ameliorating the inflammatory changes in obese adipose tissues. PMID- 19998384 TI - The red clover isoflavone irilone is largely resistant to degradation by the human gut microbiota. AB - Intestinal bacteria may influence bioavailability and physiological activity of dietary isoflavones. We therefore investigated the ability of human intestinal microbiota to convert irilone and genistein in vitro. In contrast to genistein, irilone was largely resistant to transformation by fecal slurries of ten human subjects. The fecal microbiota converted genistein to dihydrogenistein, 6' hydroxy-O-desmethylangolensin, and 2-(4-hydroxyphenyl)-propionic acid. However, considerable interindividual differences in the rate of genistein degradation and the pattern of metabolites formed from genistein were observed. Only one metabolite, namely dihydroirilone, was formed from irilone in minor amounts. In further experiments, Eubacterium ramulus, a prevalent flavonoid-degrading species of the human gut, was tested for transformation of irilone. In contrast to genistein, irilone was not converted by E. ramulus. Irilone only differs from genistein by a methylenedioxy group attached to the A-ring of the isoflavone skeleton. This substitution obviously restricts the degradability of irilone by human intestinal bacteria. PMID- 19998385 TI - Sterigmatocystin: occurrence in foodstuffs and analytical methods--an overview. AB - Sterigmatocystin (STC) is a mycotoxin produced by fungi of many different Aspergillus species. Other species such as Bipolaris, Chaetomium, Emiricella are also able to produce STC. STC producing fungi were frequently isolated from different foodstuffs, while STC was regularly detected in grains, corn, bread, cheese, spices, coffee beans, soybeans, pistachio nuts, animal feed and silage. STC shows different toxicological, mutagenic and carcinogenic effects in animals and has been recognized as a 2B carcinogen (possible human carcinogen) by International Agency for Research on Cancer. There are more than 775 publications available in Scopus (and more than 505 in PubMed) mentioning STC, but there is no summary information available about STC occurrence and analysis in food. This review presents an overview of the worldwide information on the occurrence of STC in different foodstuffs during the last 40 years, and describes the progress made in analytical methodology for the determination of STC in food. PMID- 19998386 TI - J-compensated PGSE: an improved NMR diffusion experiment with fewer phase distortions. AB - Peak distortion caused by homonuclear (1)H J-coupling is a major problem in many spin-echo-based experiments such as pulsed gradient spin-echo (PGSE) experiments. Although peak phase distortions can be lessened by the incorporation of anti phase purging sequences, the sensitivity is substantially decreased. Techniques for lessening the effect of homonuclear J-coupling evolution in spin-echo-based experiments have been investigated. Two potentially useful candidates include a J compensated inversion sequence that is efficient over a wide range of J-coupling values and a pulse sequence that refocuses homonuclear J-evolution during the spin-echo. The latter was found to work superbly on samples containing two spin (AX or AB) systems and still provided significant advantage over the standard method on samples containing more complicated spin systems. Implementation of this J-refocusing technique into a PGSE-type experiment (J-PGSE) leads to dramatic improvement of spectra and easier data analysis. The J-PGSE sequence should find applications in many diffusion studies where the PGSE-type method is required and should be a viable alternative to PGSTE especially in dilute samples due to its enhanced sensitivity. PMID- 19998387 TI - Major histocompatibility complex class I-related chains A and B (MIC A/B): a novel role in nonalcoholic steatohepatitis. AB - Stress-induced soluble major histocompatibility complex class I-related chains A/B (MIC A/B) are increased in chronic liver diseases and hepatocellular malignancy. We investigated the impact of these molecules on liver injury, apoptosis, and fibrosis in nonalcoholic steatohepatitis (NASH). Blood and liver tissue were obtained from 40 patients with NASH undergoing bariatric surgery for obesity. The control group consisted of 10 healthy individuals. We also investigated 10 patients with nonalcoholic fatty liver (NAFL). Polymerase chain reaction was used to measure messenger RNA (mRNA) transcripts of MIC A/B, natural killer cell receptor G2D (NKG2D), CD95/Fas, and tumor necrosis factor-related apoptosis-inducing ligand (TRAIL)-death receptor 5 (DR5). Apoptosis was quantified by way of terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL) (intrahepatic) and M30/M65 (systemic). Liver injury was assessed histopathologically and serologically (alanine aminotransferase/aspartate aminotransferase). Fibrosis was identified by Sirius red staining, quantitative morphometry, and alpha-smooth muscle actin and collagen 1alpha transcripts. Compared with controls, patients with NASH revealed significant increases in (1) NKG2D mRNA (13.1-fold) and MIC A/B mRNA (3.6-fold and 15.8-fold, respectively); (2) TRAIL-DR5 and CD95/Fas mRNA (2.7-fold and 3.6-fold, respectively); (3) TUNEL positive hepatocytes (4.0-fold); and (4) M30 and M65 levels (4.6-fold and 3.4 fold, respectively). We found relevant correlations between MIC protein expression rates and NAS and fibrosis stages. In contrast, NKG2D and MIC A/B transcripts were attenuated in patients with NAFL compared with NASH. Histopathologically, NASH patients revealed increased NAS scores, an accumulation of natural killer cells, and 2.7-fold increased hepatic fibrosis by quantitative morphometry. CONCLUSION: Our findings suggest an important role for MIC A/B in liver injury. Therapeutic intervention aimed at reducing MIC A/B levels may beneficially affect the progression of NASH. PMID- 19998388 TI - Derivatization of fatty alcohol ethoxylate non-ionic surfactants using 2 sulfobenzoic anhydride for characterization by liquid chromatography/mass spectrometry. AB - A derivatization procedure has been developed for the improved characterization of fatty alcohol ethoxylate non-ionic surfactants by liquid chromatography/mass spectrometry. The end hydroxyl group of each surfactant species was converted into an oxycarbonylbenzene-2-sulfonic acid group with 2-sulfobenzoic anhydride under mild conditions. The produced sulfonic acid group allows all species, including fatty alcohols and those with less than three ethoxylates, to be uniformly ionized by electrospray ionization (ESI) mass spectrometry. Both acid and base can be used as a mobile phase additive for liquid chromatography without affecting M(n) and average ethoxylate values, although ion intensities are suppressed during the ESI process. The method was used to analyze seven commercial fatty alcohol ethoxylate non-ionic surfactants, and the determined M(n) and EO values were comparable with the results obtained by NMR. The relative ratio of different fatty alcohol based ethoxylates in a sample can also be determined using the summed mass spectral data. PMID- 19998389 TI - 13C isotope effects on 1H chemical shifts: NMR spectral analysis of 13C-labelled D-glucose and some 13C-labelled amino acids. AB - The one- and two-bond (13)C isotope shifts, typically -1.5 to -2.5 ppb and -0.7 ppb respectively, in non-cyclic aliphatic systems and up to -4.4 ppb and -1.0 ppb in glucose cause effects that need to be taken into account in the adaptive NMR spectral library-based quantification of the isotopomer mixtures. In this work, NMR spectral analyses of some (13)C-labelled amino acids, D-glucose and other small compounds were performed in order to obtain rules for prediction of the (13)C isotope effects on (1)H chemical shifts. It is proposed that using the additivity rules, the isotope effects can be predicted with a sufficient accuracy for amino acid isotopomer applications. For glucose the effects were found strongly non-additive. The complete spectral analysis of fully (13)C-labelled D glucose made it also possible to assign the exocyclic proton signals of the glucose. PMID- 19998390 TI - Complete analysis of the 1H and 13C NMR spectra of diastereomeric mixtures of (R,S- and S,S-)-3,6-dimethoxy-2,5-dihydropyrazine-substituted indoles and their conformational preference in solution. AB - Complete analysis of the (1)H and (13)C NMR spectra obtained with and without a chemical shift reagent (Eu(fod)(3)), of bis-lactim ether 1 (Schollkopf auxiliary) and monosubstituted 3- or 2-{(2R,5S or 2S,5S)-5-isopropyl-3,6-dimethoxy-2,5 dihydropyrazin-2-yl]methyl}-1H-indoles is presented using gradient-selected one dimensional (1D) and two-dimensional NMR techniques, such as 1D TOCSY, 1D NOESY (DPFGSE NOE), gCOSY, NOESY, ROESY gHETCOR, gHSQC and gHMBC. The contour plot of the gCOSY spectrum of 1-10 revealed cross peaks arising from the five-bond coupling between the H2 and H5 resonances of the dihydropyrazine ring for syn- ((5)J(H2, H5) = 4-5.7 Hz) and for anti-isomers ((5)J(H2, H5) = 3.4-3.8 Hz). The magnitude of the coupling constant was utilized to distinguish between the syn- and the anti-isomers (diastereomers). The precise values of (n)J(HH) (n = 3, 4, 5, 6) coupling constants for the indole and 2,5-dihydropyrazine moieties deduced from the calculated NMR spectra were supported by 1D TOCSY and gCOSY experiments and gauge invariant atomic orbital (GIAO) calculations. The magnitude of the coupling constants ((5)J(H2, H5)) indicates that the dihydropyrazine ring exists in a boat conformation. In both isomers, the indole group adopts a 'folded' conformation in which one diastereotopic face is effectively shielded by the aromatic benzene ring of the indole. This is supported by gradient-selected 1D NOESY and 2D NOESY experiments. Theoretical calculations of the conformation were performed to support the through-space shielding effect of the aromatic indole moiety based on the DFT/GIAO calculated (1)H NMR data (chemical shifts and coupling constants) for 2-syn- and 2-anti-diastereomers in CDCl(3). PMID- 19998391 TI - Grooming patterns in Verreaux's sifaka. AB - Lemur grooming has received very little attention in the literature. Nevertheless, allogrooming in lemurs has been suggested to be fundamentally different from the grooming of anthropoids. One reason is that lemurs generally rely on oral rather than manual grooming. Lemur allogrooming has also been suggested to serve less of a social function than has been attributed to anthropoid grooming. I analyzed the allogrooming behaviors of 29 Verreaux's sifaka (Propithecus verreauxi) living in five social groups in the Kirindy Forest of Madagascar. Based upon 1,586 observation hours, I found that sifaka, like anthropoids, spend very little time mutual grooming (2+/-3%). Half of all allogrooming involved parts of the body that could have been easily groomed by the recipient, such as the limbs. Even though ectoparasite loads are expected to be greater during the rainy season, allogrooming did not increase during the rainy season. Allogrooming rates were influenced by both rank and sex, and increased by 50-100% during the mating season. The results of this study suggest that allogrooming in Verreaux's sifaka plays an important social function, even though it is performed with a toothcomb. PMID- 19998392 TI - Developing an effective community conservation program for cotton-top tamarins (Saguinus oedipus) in Colombia. AB - Developing effective conservation programs that positively impact the survival of a species while considering the needs of local communities is challenging. Here we present an overview of the conservation program developed by Proyecto Titi to integrate local communities in the conservation of Colombia's critically endangered primate, the cotton-top tamarin (Saguinus oedipus). Our comprehensive assessment of the threats effecting the long-term survival of the cotton-top tamarin allowed us to establish the primary components of our program. Proyecto Titi has three areas of emphasis: (1) scientific studies detailing the biology and long-term survival of the cotton-top tamarin, (2) conservation education programs to increase public awareness and conservation knowledge, and (3) community empowerment programs that demonstrate a valuable economic incentive to protecting wildlife and forested areas in Colombia. This integrated approach to conservation that involves local communities in activities that benefit individuals, as well as wildlife, has proven to be remarkably effective in protecting cotton-top tamarins and their forested habitat. Our bindes program, which uses small cook stoves made from clay, has demonstrated a marked reduction in the number of trees that have been harvested for firewood. Developing environmental entrepreneurs, who create products made from recycled plastic for sale in national and international markets, has had a significant impact in reducing the amount of plastic that has been littering the environment and threatening the health of wildlife, while creating a stable economic income for rural communities. Proyecto Titi has provided economic alternatives to local communities that have dramatically reduced the illegal capture of cotton-top tamarins and forest destruction in the region that has positively impacted the long-term survival of this critically endangered primate. PMID- 19998394 TI - Comparing methods of measurement: Extending the LoA by regression. AB - Method comparison studies are usually analyzed by computing limits of agreement (LoA). If only one measurement by each method is taken on each person, and the difference across the range is not constant, it has been suggested (Stat. Methods Med. Res. 1999; 8:136-160) to regress the differences on the averages and use the resulting equation to construct LoA. LoA can be converted to a prediction formula for one method given a measurement by the other. The meaning of the regression of differences on means is clarified in the framework of a proper model and prediction equations linking one method to another are devised. The performance of this model based method is evaluated against the simple approach proposed earlier and against the Deming regression. PMID- 19998393 TI - Preventive interventions among children exposed to trauma of armed conflict: a literature review. AB - Increasing research is available on the preconditions for child mental health and optimal development in traumatic conditions, whereas less is known how to translate the findings into effective interventions to help traumatized children. This literature review analyses the effectiveness of psychosocial preventive interventions and treatments and their theoretical bases among children traumatized in the context of armed conflicts (war, military violence, terrorism and refugee). The first aim is to evaluate the effectiveness of preventive interventions in preventing emotional distress and impairment and promoting optimal emotional-cognitive and social development. The second task is to analyze the nature of the underlying mechanisms for the success of preventive interventions, and the theoretical premises of the choice of intervention techniques, procedures and tools. We found 16 relevant published studies, but an examination of them revealed that only four of them had experimental designs strong enough that they could be included in the meta-analysis. While the subjective reports of the researchers suggested that systematic preventive interventions were effective in decreasing PTSD and depressive symptoms among children traumatized due to armed conflict, the more objective results of the meta-analysis and the weaknesses in designs uncovered during the meta-analysis undermine such a conclusion. Additionally, a majority of the reported preventive interventions focused only on children's biased cognitive processes and negative emotions, while only a few aimed at influencing multiple domains of child development and improving developmental functioning on emotional, social and psychophysiological levels. It is concluded that substantial additional work needs to be done in developing effective preventive interventions and treatments for children traumatized by exposure to war and violence. PMID- 19998395 TI - Zwitterionic relatives of cationic platinum group metal complexes: applications in stoichiometric and catalytic sigma-bond activation. AB - Zwitterionic platinum group metal complexes that feature formal charge separation between a cationic metal fragment and a negatively charged ancillary ligand combine the desirable reactivity profile of related cationic complexes with the broad solubility and solvent tolerance of neutral species. As such, zwitterionic complexes of this type have emerged as attractive candidates for a diversity of applications, most notably involving the breaking and/or forming of E-H and E-C sigma bonds involving a main group element E. Important advances in ancillary ligand design are documented that have enabled the construction of platinum group metal zwitterions. Also summarized are the results of stoichiometric and catalytic investigations in which the reactivity of such zwitterions and their more traditionally employed cationic relatives in sigma bond activation chemistry are compared and contrasted. PMID- 19998396 TI - Through-bond energy transfer cassettes with minimal spectral overlap between the donor emission and acceptor absorption: coumarin-rhodamine dyads with large pseudo-Stokes shifts and emission shifts. PMID- 19998397 TI - Prescription refill records as a screening tool to identify antidepressant non adherence. AB - PURPOSE: Non-adherence is a significant problem with antidepressants. Identifying patients at highest risk for discontinuing antidepressant treatment can be used to target clinical management. Accordingly, our purpose was to determine the shortest gap in medication supply that is predictive of discontinuation, while minimizing false positive results. METHODS: A retrospective cohort study of medical and prescription claims from a national health plan affiliated with i3 Innovus. Sensitivities, specificities, and positive and negative predictive values were calculated for gap lengths to assess how well they predicted discontinuation. Continuously insured individuals aged 18-65 with newly diagnosed major depression and an antidepressant prescription within 45 days of diagnosis were included. Gap length was defined as the maximum number of continuous days without medication supply during acute phase treatment. Discontinuation was defined as a continuous gap of 30 or more days between an expected refill and actual refill. RESULTS: Of 4545 eligible patients, 73% discontinued antidepressant treatment during the study period. A maximum continuous gap of 14 days had a sensitivity of 87% and a specificity of 82% for predicting discontinuation. In analyses that varied the way gaps and discontinuation were defined, gap lengths between 8 and 19 days were highly predictive of discontinuation without exceeding a 20% false positive rate. CONCLUSIONS: Based on administrative pharmacy records, screening for gaps in medication supply of at least 14 days can accurately identify four of every five patients at risk for discontinuing. This early indicator can be used to target clinical interventions. PMID- 19998399 TI - Spontaneous deracemization of disc-like molecules in the columnar phase. PMID- 19998398 TI - Wogonin inhibits osteoclast formation induced by lipopolysaccharide. AB - To evaluate the inhibitory activity of wogonin against lipopolysaccharide (LPS) induced bone resorption, we investigated the effect of wogonin on osteoclastogenesis induced by LPS. Wogonin inhibited LPS-induced osteoclastogenesis in co-cultures of mouse calvaria-derived osteoblasts and bone marrow-derived pre-osteoclasts. Wogonin also suppressed osteoclastogenesis in LPS injected mouse calvaria. In osteoblasts, the upregulation of receptor activator of nuclear factor-kappaB (RANKL) expression and the downregulation of osteoprotegerin (OPG) expression by LPS were inhibited by wogonin. Wogonin and NS 398, a COX-2 inhibitor, suppressed LPS-stimulated PGE(2) production in osteoblasts. NS-398 inhibited the effect of LPS on RANKL and OPG expression in osteoblasts. These results suggest that wogonin acts as an inhibitor of LPS induced osteoclastogenesis through downregulation of RANKL and upregulation of OPG expression via blockage of PGE(2) production. Based on these results, wogonin has potential for use as a therapeutic agent in bacteria-induced bone resorption. PMID- 19998401 TI - Hydrogen-bond networks in water clusters (H2O)20: an exhaustive quantum-chemical analysis. AB - Water aggregates allow for numerous configurations due to different distributions of hydrogen bonds. The total number of possible hydrogen-bond networks is very large even for medium-sized systems. We demonstrate that targeted ultra-fast methods of quantum chemistry make an exhaustive analysis of all configurations possible. The cage of (H(2)O)(20) in the form of the pentagonal dodecahedron is a common motif in water structures. We calculated the spatial and electronic structure of all hydrogen-bond configurations for three systems: idealized cage (H(2)O)(20) and defect cages with one or two hydrogen bonds broken. More than 3 million configurations studied provide unique data on the structure and properties of water clusters. We performed a thorough analysis of the results with the emphasis on the cooperativity in water systems and the structure property relations. PMID- 19998400 TI - The thioesterase Bhp is involved in the formation of beta-hydroxytyrosine during balhimycin biosynthesis in Amycolatopsis balhimycina. AB - The putative hydrolase gene bhp from the balhimycin biosynthetic gene cluster has been cloned and overexpressed in Escherichia coli. The corresponding enzyme Bhp was purified to homogeneity by nickel-chelating chromatography and characterized. Although Bhp has sequence similarities to hydrolases with "haloperoxidase"/perhydrolase activity, it did not show any enzymatic activity with standard "haloperoxidase"/perhydrolase substrates (e.g., monochlorodimedone and phenol red), nonspecific esterase substrates (such as p-nitrophenyl acetate, p-nitrophenyl phosphate and S-thiophenyl acetate) or the model lactonase substrate dihydrocoumarin. However, Bhp could be shown to catalyse the hydrolysis of S-beta-hydroxytyrosyl-N-acetyl cysteamine thioester (beta-OH-Tyr-SNAC) with 15 times the efficiency of S-L-tyrosyl-N-acetyl cysteamine thioester (L-Tyr-SNAC). This is in agreement with the suggestion that Bhp is involved in balhimycin biosynthesis, during which it was supposed to catalyse the hydrolysis of beta-OH Tyr-S-PCP (PCP=peptidyl carrier protein) to free beta-hydroxytyrosine (beta-OH Tyr) and strongly suggests that Bhp is a thioesterase with high substrate specificity for PCP-bound beta-OH-Tyr and not a "haloperoxidase"/perhydrolase or nonspecific esterase. PMID- 19998402 TI - Oriented electric fields accelerate Diels-Alder reactions and control the endo/exo selectivity. AB - Herein we demonstrate that an external electric field (EEF) acts as an accessory catalyst/inhibitor for Diels-Alder (DA) reactions. When the EEF is oriented along the "reaction axis" (the coordinate of approach of the reactants in the reaction path), the barrier of the DA reactions is lowered by a significant amount, equivalent to rate enhancements by 4-6 orders of magnitude. Simply flipping the EEF direction has the opposite effect, and the EEF acts as an inhibitor. Additionally, an EEF oriented perpendicular to the "reaction axis" in the direction of the individual molecule dipoles can change the endo/exo selectivity, favouring one or the other depending on the positive/negative directions of the EEF vis-a-vis the individual molecular dipole. At some critical value of the EEF along the "reaction axis", there is a crossover to a stepwise mechanism that involves a zwitterionic intermediate. The valence bond diagram model is used to comprehend these trends and to derive a selection rule for EEF effects on chemical reactions: an EEF aligned in the direction of the electron flow between the reactants will lower the reaction barrier. It is shown that the exo/endo control by the EEF is not associated with changes in secondary orbital interactions. PMID- 19998403 TI - Voltage-induced payload release and wettability control on TiO2 and TiO2 nanotubes. PMID- 19998404 TI - Kinetics of the self-aggregation and film formation of poly-L-proline at high temperatures explored by circular dichroism spectroscopy. AB - Poly-L-proline has been used as a model system for various purposes over a period of more than 60 years. Its relevance among the protein/peptide community stems from its use as a reference system for determining the conformational distributions of unfolded peptides and proteins, its use as a molecular ruler, and from the pivotal role of proline residues in conformational transitions and protein-protein interactions. While several studies indicate that polyproline can aggregate and precipitate in aqueous solution, a systematic study of the aggregation process is still outstanding. We found, by means of UV-circular dichroism and IR measurements, that polyproline is predominantly monomeric at room temperature at millimolar concentrations. Upon heating, the polypeptide stays in its monomeric state until the temperature reaches a threshold of ca. 60 degrees C. At higher temperatures, the peptide aggregates as a film on the inside surface of the employed cuvette. The process proceeds on a time scale of 10(3) s and can best be described by a bi-exponential relaxation function. The respective CD and IR spectra are qualitatively different from the canonical spectra of polyproline in aqueous solution, and are indicative of a highly packed state. PMID- 19998406 TI - Radiation inactivation of galactose oxidase, a monomeric enzyme with a stable free radical. AB - To determine the radiation sensitivity of galactose oxidase, a 68 kDa monomeric enzyme containing a mononuclear copper ion coordinated with an unusually stable cysteinyl-tyrosine (Cys-Tyr) protein free radical. Both active enzyme and reversibly rendered inactive enzyme were irradiated in the frozen state with high energy electrons. Surviving polypeptides and surviving enzyme activity were analyzed by radiation target theory giving the radiation sensitive mass for each property. In both active and inactive forms, protein monomer integrity was lost with a single radiation interaction anywhere in the polypeptide, but enzymatic activity was more resistant, yielding target sizes considerably smaller than that of the monomer. These results suggest that the structure of galactose oxidase must make its catalytic activity unusually robust, permitting the enzymatic properties to survive in molecules following cleavage of the polymer chain. Radiation target size for loss of monomers yielded the mass of monomers indicating a polypeptide chain cleavage after a radiation interaction anywhere in the monomer. Loss of enzymatic activity yielded a much smaller mass indicating a robust structure in which catalytic activity could be expressed in cleaved polypeptides. PMID- 19998405 TI - Human tissue transglutaminase is inhibited by pharmacologic and chemical acetylation. AB - Human tissue transglutaminase (TGM2) is implicated in the pathogenesis of several neurodegenerative disorders including Alzheimer's, Parkinson's and expanded polyglutamine (polyQ) diseases. TGM2 promotes formation of soluble and insoluble high molecular weight aggregates by catalyzing a covalent linkage between peptide bound Q residues in polyQ proteins and a peptide-bound Lys residue. Therapeutic approaches to modulate the activity of TGM2 are needed to proceed with studies to test the efficacy of TGM2 inhibition in disease processes. We investigated whether acetylation of Lys-residues by sulfosuccinimidyl acetate (SNA) or aspirin (ASA) would alter the crosslinking activity of TGM2. Acetylation by either SNA and/or ASA resulted in a loss of >90% of crosslinking activity. The Lys residues that were critical for inhibition were identified by mass spectrometry as Lys(444), Lys(468), and Lys(663). Hence, acetylation of Lys-residues may modulate the enzymatic function of TGM2 in vivo and offer a novel approach to treatment of TGM2 mediated disorders. PMID- 19998407 TI - De novo structure generation using chemical shifts for proteins with high sequence identity but different folds. AB - Proteins with high-sequence identity but very different folds present a special challenge to sequence-based protein structure prediction methods. In particular, a 56-residue three-helical bundle protein (GA(95)) and an alpha/beta-fold protein (GB(95)), which share 95% sequence identity, were targets in the CASP-8 structure prediction contest. With only 12 out of 300 submitted server-CASP8 models for GA(95) exhibiting the correct fold, this protein proved particularly challenging despite its small size. Here, we demonstrate that the information contained in NMR chemical shifts can readily be exploited by the CS-Rosetta structure prediction program and yields adequate convergence, even when input chemical shifts are limited to just amide (1)H(N) and (15)N or (1)H(N) and (1)H(alpha) values. PMID- 19998408 TI - Bile salt sequestration induces hepatic de novo lipogenesis through farnesoid X receptor- and liver X receptor alpha-controlled metabolic pathways in mice. AB - Diabetes is characterized by high blood glucose levels and dyslipidemia. Bile salt sequestration has been found to improve both plasma glycemic control and cholesterol profiles in diabetic patients. Yet bile salt sequestration is also known to affect triglyceride (TG) metabolism, possibly through signaling pathways involving farnesoid X receptor (FXR) and liver X receptor alpha (LXRalpha). We quantitatively assessed kinetic parameters of bile salt metabolism in lean C57Bl/6J and in obese, diabetic db/db mice upon bile salt sequestration using colesevelam HCl (2% wt/wt in diet) and related these to quantitative changes in hepatic lipid metabolism. As expected, bile salt sequestration reduced intestinal bile salt reabsorption. Importantly, bile salt pool size and biliary bile salt secretion remained unchanged upon sequestrant treatment due to compensation by de novo bile salt synthesis in both models. Nevertheless, lean and db/db mice showed increased, mainly periportally confined, hepatic TG contents, increased expression of lipogenic genes, and increased fractional contributions of newly synthesized fatty acids. Lipogenic gene expression was not induced in sequestrant treated Fxr(-/-) and Lxralpha(-/-) mice compared with wild-type littermates, in line with reports indicating a regulatory role of FXR and LXRalpha in bile salt mediated regulation of hepatic lipid metabolism. CONCLUSION: Bile salt sequestration by colesevelam induces the lipogenic pathway in an FXR- and LXRalpha-dependent manner without affecting the total pool size of bile salts in mice. We speculate that a shift from intestinal reabsorption to de novo synthesis as source of bile salts upon bile salt sequestration affects zonation of metabolic processes within the liver acinus. PMID- 19998409 TI - Farnesoid X receptor alleviates age-related proliferation defects in regenerating mouse livers by activating forkhead box m1b transcription. AB - Elucidating the mechanism of liver regeneration could lead to life-saving therapy for a large number of patients, especially elderly patients, after segmental liver transplantation or resection of liver tumors. The forkhead box m1b (Foxm1b) transcription factor is required for normal liver regeneration. Here we report that Foxm1b is the first direct farnesoid X receptor (FXR) target gene known to be involved in cell cycle regulation and that aging regenerating livers have delayed activation of FXR, which results in defective induction of Foxm1b and thereby contributes to defective liver regeneration. An inverted repeat 0 (IR-0) FXR response element, acting as an enhancer in intron 3 of the Foxm1b gene, was identified by a combination of transcriptional reporter, electrophoretic mobility shift, and chromatin immunoprecipitation assays. Diminished FXR binding to the IR 0 element was found in aging regenerating livers. FXR activation by a novel ligand in aging livers induced Foxm1b expression and elevated hepatocyte DNA replication to about 70% of the levels found in young regenerating livers, which were specifically suppressed by hepatic expression of anti-Foxm1b short hairpin RNA. CONCLUSION: Our results have revealed Foxm1b as the first known direct FXR target gene involved in cell cycle regulation and have demonstrated that defective activation of FXR could be an intrinsic defect in aging regenerating livers. Activation of FXR alone is largely able to alleviate age-related liver regeneration defects. These findings highlight FXR as a potential target of drug design for promoting liver regeneration in older subjects. PMID- 19998410 TI - PGE2 inhibits MMP expression by suppressing MKK4-JNK MAP kinase-c-JUN pathway via EP4 in human articular chondrocytes. AB - Prostaglandin E2 (PGE2) is one of pro-inflammatory mediators. PGE2 maintains the homeostasis of many organs including articular cartilage, and a previous report showed that continuous inhibition of PGE2 accelerates the progression of osteoarthritis (OA). While PGE2 inhibits matrix metalloprotease (MMP) expression in several types of cells, little is known on direct effects of PGE2 on MMP expression in articular chondrocytes. The objective of this study was to investigate direct effects of PGE2 on IL-1beta-induced MMP-1 and MMP-13 expression and the intracellular signaling in articular chondrocytes. PGE2 showed inhibitory effects on IL-1beta-induced MMP-1 and MMP-13 expression demonstrated by immunoblotting both in OA and normal chondrocytes, which was further confirmed by enzyme-linked immunosorbent assay and immunohistochemistry of explant cultures of articular cartilages. An EP4 agonist, ONO-AE1-329, mimicked the inhibitory effect of PGE2, while an EP4 antagonist, ONO-AE3-208, blocked the effects. PGE2 suppressed the phosphorylation of JNK and ERK MAP kinases, but only knockdown of JNK by specific siRNA mimicked the effect of PGE2. PGE2 further inhibited the phosphorylation of MKK4 without suppression of MKK7 phosphorylation, and of c-JUN to decrease expression levels of MMP-1 and MMP-13. These results demonstrate that PGE2 inhibits IL-1beta-induced MMP-1 and MMP-13 productions via EP4 by suppressing MKK4-JNK MAP kinase-c-JUN pathway. PMID- 19998411 TI - Methylation of histone H3 and H4 by PRMT5 regulates ribosomal RNA gene transcription. AB - In an effort to understand the epigenetic regulation of ribosomal RNA gene (rDNA) expression we have previously demonstrated the role of DNA methyltransferases and methyl CpG binding proteins in rRNA synthesis. Here, we studied the role of protein arginine methyltransferase PRMT5 and the two methylated histones H3R8Me2 and H4R3Me2, in rDNA expression in Epstein Barr virus- transformed primary B cells (LCLs) and in HeLa cells responding to serum-regulated growth. Chromatin immunoprecipitation assay showed that histones H3 and H4 associated with rRNA promoters were differentially methylated at arginine residues 8 and 3, respectively, depending on its transcriptional activity. Association of PRMT5 and methylated H3 with the unmethylated promoters in resting B-cells was significantly reduced in rapidly growing LCLs. Unlike PRMT5 and H3R8Me2, histone H4 associated with both methylated and unmethylated rRNA promoters in resting B cells was methylated at the R3 residue. However, a dramatic decrease in R3 methylation of H4 recruited to the unmethylated rRNA promoters was observed in LCLs while it remained unaltered in the fraction bound to the methylated promoters. Differential interaction of PRMT5 and methylation of H3 and H4 associated with the rRNA promoters was also observed when serum starved HeLa cells were allowed to grow in serum replenished media. Ectopic expression of PRMT5 suppressed activity of both unmethylated and methylated rRNA promoter in transient transfection assay whereas siRNA mediated knockdown of PRMT5 increased rRNA synthesis in HeLa cells. These data suggest a key role of PRMT5 and the two methylated histones in regulating rRNA promoter activity. PMID- 19998412 TI - A bipartite butyrate-responsive element in the human calretinin (CALB2) promoter acts as a repressor in colon carcinoma cells but not in mesothelioma cells. AB - The short-chain fatty acid butyrate plays an essential role in colonic mucosa homeostasis through the capacity to block the cell cycle, regulate differentiation and to induce apoptosis. The beneficial effect of dietary fibers on preventing colon cancer is essentially mediated through butyrate, derived from luminal fermentation of fibers by intestinal bacteria. In epithelial cells of the colon, both in normal and colon cancer cells, the expression of several genes is positively or negatively regulated by butyrate likely through modulation of histone acetylation and thereby affecting the transcriptional activity of genes. Calretinin (CALB2) is a member of the EF-hand family of Ca(2+)-binding proteins and is expressed in a majority of poorly differentiated colon carcinoma and additionally in mesothelioma of the epithelioid and mixed type. Since CALB2 is one of the genes negatively regulated by butyrate in colon cancer cells and butyrate decreases calretinin protein expression levels in those cells, we investigated whether expression is regulated via putative butyrate-responsive elements (BRE) in the human CALB2 promoter. We identified two elements that act as butyrate-sensitive repressors in all colon cancer cell lines tested (CaCo-2, HT-29, Co-115/3). In contrast, in cells of mesothelial origin, MeT-5A and ZL34, the same two elements do not operate as butyrate-sensitive repressors and calretinin expression levels are insensitive to butyrate indicative of cell type specific regulation of the CALB2 promoter. Calretinin expression in colon cancer cells is negatively regulated by butyrate via a bipartite BRE flanking the TATA box and this may be linked to butyrate's chemopreventive activity. PMID- 19998413 TI - PPAR-beta facilitating maturation of hepatic-like tissue derived from mouse embryonic stem cells accompanied by mitochondriogenesis and membrane potential retention. AB - Relatively little is known about mitochondria metabolism in differentiating embryonic stem (ES) cells. Present research focused on several elements of cellular energy metabolism in hepatic-like tissue derived from mouse ES cells. We demonstrated that mitochondrial location patterns and mitochondrial membrane potential (DeltaPsi(m)) existed in subsequent differentiation of the tissue. Mitochondriogenesis appeared at the early stage and kept a normal DeltaPsi(m) in differentiated mature hepatocytes. Peroxisome proliferator-activated receptor alpha (PPAR-alpha) expression was transitorily increased at the beginning, and kept a relatively low level later, which accompanied by expression of PPAR-gamma coactivator (PGC)-1alpha, a master regulator of mitochondrial biogenesis. PPAR beta expression showed robust up-regulation in the late differentiation course. Enhanced co-expressions of PPAR-beta and albumin with catalysis of UDP glucuronosyltransferases (UGTs) were observed at mature stage. While PPAR-gamma expression changed little before and after differentiation. Mitochondriogenesis could be accelerated by PPAR-alpha specific agonist WY14643 and abolished by its antagonist GW6471 at the early stage. Neither of them affected mitochondrial DeltaPsi(m) and albumin generation in the differentiated hepatocytes. Furthermore, maturation of hepatic-like tissue and mitochondriogenesis in hepatocyte could be efficiently stimulated by PPAR-beta specific agonist L165041 and abolished by PPAR-beta specific antagonist GSK0660, but not affected by PPAR gamma specific agonist GW1929. In conclusion, the derived hepatic tissue morphologically possessed cellular energy metabolism features. PPAR-alpha seemed only necessary for early mitochondriogenesis, while less important for DeltaPsi(m) retention in the mature tissue derived. The stimulation of PPAR-beta but not -gamma enhanced hepatogenesis, hepatocytes maturation, and mitochondriogenesis. PPAR-beta took an important role in cellular energy metabolism of hepatogenesis. PMID- 19998414 TI - Summary of recommendations for primate conservation education programs. PMID- 19998415 TI - MicroRNAs align with accessible sites in target mRNAs. AB - The importance of microRNAs (miRs) in control of gene expression is now clearly recognized. While individual microRNAs are thought to target hundreds of disparate mRNAs via imperfect base pairing, little is known about the characteristics of miR target sites. Here we show that the miRs can be aligned with empirically identified accessible sites in a target RNA (Cytokeratin 19, KRT), and that some of the aligned miRs functionally down-regulate KRT expression post-transcriptionally. We employed an RNase-H-based random library selection protocol to identify accessible sites in KRT RNA. We then aligned the Sanger Institute database collection of human miRs to KRT mRNA, and also aligned them using the web-based MicroInspector program. Most miRs aligned with the accessible sites identified empirically; those not aligned with the empirically identified sites also functioned effectively in RNase-H-based assays. Similar results were obtained with a second target RNA (Mammoglobin). Transient transfection assays established that some of the miRs which aligned with KRT significantly down regulated it at the protein level, with no effect on RNA level. The functionally effective miRs aligned within the coding region of KRT, whereas a number of miRs which aligned with the 3'-untranslated region did not produce down-regulation. PMID- 19998416 TI - Transient expression of serotonin 5-HT4 receptors in the mouse developing thalamocortical projections. AB - The serotonin 5-HT(4) receptor (5-HT(4)-R) is an unusually complex G-protein coupled receptor that is likely to play important roles in brain development and that may underlie the comorbidity of central and peripheral abnormalities in some developmental disorders. We studied the expression of 5-HT(4)-Rs in the developing mouse forebrain at embryonic days 13, 15, 17, and at postnatal days 3 and 14 by using immunohistochemistry, tract tracing, and quantitative RT-PCR. The developing thalamocortical projections transiently expressed 5-HT(4)-Rs in the embryonic brain and the 5-HT(4)-R expression in the forebrain changed from axonal to somatic around birth. From embryonic days 13-17, the forebrain mRNA levels of the 5-HT(4(a))-R and 5-HT(4(b))-R splice variants increased nine- and fivefold, respectively, whereas the levels of the 5-HT(4(e))-R and 5-HT(4(f))-R variants remained relatively low throughout the studied period of embryonic development. These results suggest that during development 5-HT(4)-R expression undergoes a dynamic regulation and that this regulation may be important for the normal development of sensory and limbic processing. PMID- 19998417 TI - The efficacy of Echinacea in a 3-D tissue model of human airway epithelium. AB - We evaluated the antirhinovirus efficacy of a standardized preparation of Echinacea purpurea (Echinaforce) in a 3-dimensional organotypic model of normal human airway epithelium (EpiAirway tissue). Individual replicate tissue samples, maintained as inserts in culture for 3 days or 3 weeks, were infected with rhinovirus type 1A (RV1A), Echinacea alone, a combination of the two, or medium only. None of the treatments affected the histological appearance or integrity of the tissues, all of which maintained a high level of cell viability and preservation of cilia. RV infection resulted in increased mucopolysaccharide inclusions in the goblet cells, but this feature was reversed by Echinacea treatment. This result was confirmed by measurements of mucin secretion, which was stimulated by RV but reversed by Echinacea, suggesting that mucus production during colds could be ameliorated by Echinacea. We did not find evidence of virus replication, although the RV-infected tissues secreted substantial amounts of the pro-inflammatory cytokines IL-6 and IL-8 (CXCL8), and this response was reversed by Echinacea treatment. These results confirmed previous findings derived from studies of bronchial and lung epithelial cell lines, namely, that RV infection results in a substantial inflammatory response in the absence of virus replication. PMID- 19998418 TI - The ethyl acetate extract of Phellinus linteus grown on germinated brown rice induces G0/G1 cell cycle arrest and apoptosis in human colon carcinoma HT29 cells. AB - It is well known that Phellinus linteus has a variety of biological functions, such as antitumor and immunomodulating activities. In our previous studies, we developed a P. linteus grown on germinated brown rice (PBR) and found that organic solvent extracts of PBR possessed immunomodulating activity to regulate a balance of cytokine network in mice. The components of PBR are ergosterol peroxide, gamma-aminobutyric acid (GABA) and Beta-glucan. In this study, we demonstrate that an organic solvent extract of P. linteus grown on PBR induced apoptotic cell death through the induction of G(0)/G(1) arrest of cell cycle and the apoptosis via DNA fragmentation in human colon carcinoma HT-29 cells. Cell death induced by the extract of P. linteus grown on PBR was shown to be associated with the upregulation of p21(CIP1/WAF1), the downregulation of cyclin D1, anti-apoptotic protein, Bcl-2, the release of cytochrome c, and the activation of caspase-9, caspase-3 and caspase-8. This study suggests that the ethyl acetate extract of P. linteus grown on PBR induces apoptosis accompanied by cell cycle arrest at G(0)/G(1) phase and regulates apoptosis-regulatory proteins, which may be applicable to anticancer therapy. PMID- 19998419 TI - Current practice in injection augmentation of the vocal folds: indications, treatment principles, techniques, and complications. AB - OBJECTIVES/HYPOTHESIS: To identify contemporary indications, treatment principles, technique, injection materials, complications, and success rates of vocal fold injection augmentation. STUDY DESIGN: Multi-institutional retrospective review. METHODS: Records of patients undergoing injection augmentation at seven university medical centers from July 2007 through June 2008 were reviewed for information regarding diagnosis, unilateral or bilateral injection, route of injection, anesthesia, treatment site (office or operating room), material used, reason for technique selected, and technical success. RESULTS: In 12 months, 460 injections were performed, 236 (51%) in awake, unsedated patients, and 224 (49%) under general anesthesia. Indications included vocal fold paralysis (248; 54%), paresis (97; 21%), atrophy (68; 15%) and scar (47; 10%). Scar was more likely to be treated in the operating room (P = .000052). In awake patients, 112 (47%) injections were performed by transcricothyroid approach, 55 (23%) by peroral approach, 49 (21%) by transthyrohyoid membrane approach, and 20 (8%) by transthyroid cartilage approach. Neither technical success rate (99% vs. 97%) nor complication rate (3% vs. 2%) differed between awake and asleep techniques. The most common materials in the clinic setting were methylcellulose (35%), bovine collagen (28%), and calcium hydroxylapatite (26%); in the operating room these were calcium hydroxylapatite (36%) and methylcellulose (35%). Calcium hydroxylapatite was more likely to be used under general anesthesia (P = .019). Five-year data show that the use of injection in the awake patient rose from 11% to 43% from 2003 to 2008. CONCLUSIONS: Injection augmentation remains a safe, effective, and clinically practical treatment with a high rate of success, whether performed in the awake or asleep patient. The rapid adoption of awake injection over the past 5 years speaks to its clinical utility. Complication rates are low and equivalent to those under general anesthesia. Otolaryngologists continue to use a variety of techniques and materials to treat a range of conditions of glottic insufficiency. PMID- 19998420 TI - Contact endoscopy for the evaluation of the pharyngeal and laryngeal mucosa. AB - OBJECTIVES: Contact endoscopy is a noninvasive tool that allows in vivo and in situ examination of superficial mucosa. Its use for early diagnosis of cancerous lesions of the oropharynx and larynx has not been evaluated. The aim of the study was to validate contact endoscopy for the examination of pharyngeal and laryngeal mucosa. STUDY DESIGN: Prospective clinical study. METHODS: Superficial cells of the mucosa were stained with methylene blue and examined with contact endoscopes. The documented images were assessed by a cytopathologist and by an otolaryngologist independently for each patient. Biopsies for histopathological examination of the area were performed and correlated with contact endoscopic findings of both examiners. RESULTS: Of the 42 examined specimen, 32 (76.2%) showed benign changes in the histological analysis. Squamous cell carcinoma was revealed in 10 specimen (23.8%). Using contact endoscopy, the cytopathologist accurately identified 90.6% of the benign findings (29 of 32); however, only seven of 10 (70%) carcinomas were correctly categorized. In comparison, the otolaryngologist made a correct diagnosis in 93.75% (30 of 32) of the benign and in 90% (nine of 10) of the malignant cases. Thus, a sensitivity of 90% and a specificity of 93.75% can be achieved by contact endoscopy. CONCLUSIONS: Contact endoscopy offers valuable support for the evaluation of oropharyngeal, hypopharyngeal, and laryngeal mucosa. Contact endoscopy can be a useful contribution to rapid intraoperative evaluation of mucosal alterations for early diagnosis of tumors and might reduce diagnostic biopsy sampling. Even so, it does not replace biopsy sampling. PMID- 19998421 TI - Combined antegrade and retrograde esophageal dilation for head and neck cancer related complete esophageal stenosis. AB - OBJECTIVES/HYPOTHESIS: Assess the safety and efficacy of combined antegrade and retrograde esophageal dilation (CARD) for complete esophageal stenosis following head and neck cancer (HNC) treatment. Review HNC dysphagia management. STUDY DESIGN: Retrospective review of all patients undergoing CARD following HNC treatment between May 2001 and September 2008. METHODS: Forty-five patients were identified for review. Parameters assessed included: ability to obtain intraoperative esophageal patency, complications, number of dilations required, diet, and gastric tube (GT) status. Factors associated with dilation failure were analyzed. RESULTS: Intraoperative esophageal patency was obtained in 91% of patients. Median number of all dilations per patient was three. Median number of CARDs per patient was one. Resumption of oral intake occurred in 36/45 (80%). Diet results included: regular or soft diet 32/45 (71%), GT removal 27/45 (60%), and GT dependence with nothing by mouth 9/45 (20%). Laryngeal and pharyngeal stenosis, radionecrosis, tracheotomy dependence, and elongated stenosis were associated with dilation failure. Complications occurred in 18/63 (29%) CARD procedures: eight pneumomediastinum, seven GT site problems, two esophageal perforations, and one pharyngeal infection. All complications resolved spontaneously or with minimal interventions. CONCLUSIONS: CARD was safe and effective. Intraoperative patency was achieved in 91% of patients. Eighty percent resumed oral intake. The majority of patients had their GTs removed and resumed a soft or regular diet. Dilation failure was associated with laryngeal, pharyngeal, and excessively long esophageal stenosis, often resulting from radionecrosis. Complications were minor. CARD should be considered before relegating patients with complete esophageal stenosis to chronic GT dependence or subjecting them to laryngopharyngo esophagectomy. PMID- 19998423 TI - Prospects for the automation of analysis and interpretation of flow cytometric data. PMID- 19998422 TI - SLC26A4 genotype, but not cochlear radiologic structure, is correlated with hearing loss in ears with an enlarged vestibular aqueduct. AB - OBJECTIVES/HYPOTHESIS: Identify correlations among SLC26A4 genotype, cochlear structural anomalies, and hearing loss associated with enlargement of the vestibular aqueduct (EVA). STUDY DESIGN: Prospective cohort survey, National Institutes of Health, Clinical Center, a federal biomedical research facility. METHODS: Eighty-three individuals, 11 months to 59 years of age, with EVA in at least one ear were studied. Correlations among pure-tone hearing thresholds, number of mutant SLC26A4 alleles, and the presence of cochlear anomalies detected by computed tomography or magnetic resonance imaging were examined. RESULTS: Linear mixed-effects model indicated significantly poorer hearing in ears with EVA in individuals with two mutant alleles of SLC26A4 than in those with EVA and a single mutant allele (P = .012) or no mutant alleles (P = .007) in this gene. There was no detectable relationship between degree of hearing loss and the presence of structural cochlear anomalies. CONCLUSIONS: The number of mutant alleles of SLC26A4, but not the presence of cochlear anomalies, has a significant association with severity of hearing loss in ears with EVA. This information will be useful for prognostic counseling of patients and families with EVA. PMID- 19998424 TI - Empowering the disempowered: harm reduction with racial/ethnic minority clients. AB - Harm-reduction principles, such as empowerment of clients and goals for moderation, align well with the cultural worldviews of many clients of color. Empirically supported harm-reduction strategies often work well with ethnic minority clients. However, clients of color require special clinical considerations. A case study illustrates the use of the community participatory model, which combines harm reduction, cultural sensitivity, and community support. Treatment services can be provided under one roof to eliminate barriers to seeking and receiving services. Harm-reduction therapists can carefully assess and diagnose clients of color to minimize the potential shortcomings and cultural biases in assessment tools. Therapy will include the family and incorporate traditional practices as desired by clients. Use of these methods will empower and support clients of color as they seek their treatment goals. PMID- 19998425 TI - Mindfulness intervention for child abuse survivors. AB - Twenty-seven adult survivors of childhood sexual abuse participated in a pilot study comprising an 8-week mindfulness meditation-based stress reduction (MBSR) program and daily home practice of mindfulness skills. Three refresher classes were provided through final follow-up at 24 weeks. Assessments of depressive symptoms, post-traumatic stress disorder (PTSD), anxiety, and mindfulness, were conducted at baseline, 4, 8, and 24 weeks. At 8 weeks, depressive symptoms were reduced by 65%. Statistically significant improvements were observed in all outcomes post-MBSR, with effect sizes above 1.0. Improvements were largely sustained until 24 weeks. Of three PTSD symptom criteria, symptoms of avoidance/numbing were most greatly reduced. Compliance to class attendance and home practice was high, with the intervention proving safe and acceptable to participants. These results warrant further investigation of the MBSR approach in a randomized, controlled trial in this patient population. PMID- 19998429 TI - Reversible switching of a micelle-to-vesicle transition by compressed CO2. AB - The study of the micelle-to-vesicle transition (MVT) is of great importance from both theoretical and practical points of view. Herein, we studied the effect of compressed CO(2) on the aggregation behavior of dodecyltrimethylammonium bromide (DTAB)/sodium dodecyl sulfate (SDS) mixed surfactants in aqueous solution by means of direct observation, turbidity and conductivity measurements, steady state fluorescence, time-resolved fluorescence quenching (TRFQ), fluorescence quantum yield, and template methods. Interestingly, all these approaches showed that compressed CO(2) could induce the MVT in the surfactant system, and the vesicles returned to the micelles simply by depressurization; that is, CO(2) can be used to switch the MVT reversibly by controlling pressure. Some other gases, such as methane, ethylene, and ethane, could also induce the MVT of the surfactant solution. A possible mechanism is proposed on the basis of the packing parameter theory and thermodynamic principles. It is shown that the mechanism of the MVT induced by a nonpolar gas is different from the MVT induced by polar and electrolyte additives. PMID- 19998427 TI - E-cadherin expression in male urethral smears and correlation with PCR-based detection of human papillomavirus infection. AB - We performed cytological examination of urethral brushing to study aberrant E Cadherin expression as a possible marker for papillomavirus in cytological samples. A total of 30 cytobrush male urethral smears were examined E-cadherin expression and human papillomavirus (hpv) was confirmed using PCR-DNA. The age range was 19-36 years (mean: 27 years), 17 (56.7%) cases corresponded to low-risk HPV and 13 (43.3%) cases were high-risk HPV. The mean age ranges were 25.77 +/- 5.90 years for high-risk HPV and 26.77 +/- 4.31 years for low-risk HPV. Pap smears showed dyskeratosis in 23 (76.7%) cases, koilocytes in 13 (43.3%) cases, infection or bacterial background in 14 (43.7%) cases, suggestive changes of Gardnerella infection in 7 (23.3%) cases, and Chlamydia in 3 (10%) cases. Immunohistochemistry was positive for membrane E-Cadherin; there was weak expression in 13 (43.3%) cases, moderate expression in 11 (36.7%) cases (P = 0.109) (Figs. 3a and b), and strong expression in 6 (20%) cases. There were a statistically significant correlations between E-Cadherin expression and koilocytes (P = 0.007), individual cell dyskeratosis (P = 0.041), and HPV risk (P = 0.000). We concluded that the loss of E-Cadherin membrane expression was greater in high-risk HPV cases and was associated with individual cell dyskeratosis features and koilocytes. There were statistically significant correlations between E-Cadherin expression and dyskeratosis (P = 0.043), koilocytes (P = 0.007), and type of HPV (P = 0.000). Using male urethral smears to test for the loss of E-Cadherin membrane expression is simple, rapid, specific and more sensitive than conventional morphologic observations. We concluded that E-Cadherin can be used to discriminate between high- and low-risk papillomavirus in urethral cytologic specimens. PMID- 19998430 TI - Pectin-derived porous materials. AB - Porous forms of pectin, a major industrial waste biomass polysaccharide, have been prepared by aqueous phase expansion routes (S(BET)>200 m(2) g(-1); V(pore)>0.80 cm(3) g(-1)). It was demonstrated that the aqueous phase acidity crucially influenced the properties of the porous pectin form. Preparation route selection allows direction of material textural and morphological properties, thought to be the result of polysaccharide configuration, and methyl ester group hydrolysis, believed to alter the lowest energy accessible metastable polysaccharide state during gel recrystallisation. The resulting low density amorphous powders or mouldable monoliths (rho(powder) approximately 0.20 g cm( 3)/rho(monolith) approximately 0.07 g cm(-3)) can be directly transformed by thermal carbonisation into low density mesoporous carbonaceous materials (e.g. rho approximately 0.27 g cm(-3) (T(p)=550 degrees C)), which possess textural and nanoscale material morphology reflective of the porous pectin precursor employed. Acidic gelation promotes methyl ester groups hydrolysis of the polysaccharide structure, generating carbons with unusual interdigitated rod-like nanoscale morphology. Importantly, the materials presented herein are produced directly from the parent porous pectin material, without the need for additive catalyst (or template) to yield highly mesoporous products (e.g. V(meso) > or = 0.45 cm(3) g(-1); polydispersity (PD)>10 nm), with accessible tuneable functionally rich surfaces. Due to the high mesoporosity (>85%), materials have potential application in chromatography, heterogeneous catalysis and large molecule adsorption strategies. PMID- 19998431 TI - The effect of heavy atoms on photoinduced electron injection from nonthermalized and thermalized donor states of M(II)-polypyridyl (M=Ru/Os) complexes to nanoparticulate TiO(2) surfaces: an ultrafast time-resolved absorption study. AB - We have synthesized ruthenium(II)- and osmium(II)-polypyridyl complexes ([M(bpy)(2)L](2+), in which M=Os(II) or Ru(II), bpy=2,2'-bipyridyl, and L=4-(2,2' bipyridinyl-4-yl)benzene-1,2-diol) and studied the interfacial electron-transfer process on a TiO(2) nanoparticle surface using femtosecond transient-absorption spectroscopy. Ruthenium(II)- and osmium(II)-based dyes have a similar molecular structure; nevertheless, we have observed quite different interfacial electron transfer dynamics (both forward and backward). In the case of the Ru(II)/TiO(2) system, single-exponential electron injection takes place from photoexcited nonthermalized metal-to-ligand charge transfer (MLCT) states. However, in the case of the Os(II)/TiO(2) system, electron injection takes place biexponentially from both nonthermalized and thermalized MLCT states (mainly (3)MLCT states). Larger spin-orbit coupling for the heavier transition-metal osmium, relative to that of ruthenium, accounts for the more efficient population of the (3)MLCT states in the Os(II)-based dye during the electron-injection process that yields biexponential dynamics. Our results tend to suggest that appropriately designed Os(II)-polypyridyl dye can be a better sensitizer molecule relative to its Ru(II) analogue not only due to much broader absorption in the visible region of the solar-emission spectrum, but also on account of slower charge recombination. PMID- 19998433 TI - RuCl(3)x H(2)O-catalyzed direct arylation of arenes with aryl chlorides in the presence of triphenylphosphine. PMID- 19998432 TI - A chiral pyrrolic tripodal receptor enantioselectively recognizes beta-mannose and beta-mannosides. PMID- 19998434 TI - Adsorption of CO(2), CH(4), and N(2) on zeolitic imidazolate frameworks: experiments and simulations. AB - Experimental measurements and molecular simulations were conducted for two zeolitic imidazolate frameworks, ZIF-8 and ZIF-76. The transferability of the force field was tested by comparing molecular simulation results of gas adsorption with experimental data available in the literature for other ZIF materials (ZIF-69). Owing to the good agreement observed between simulation and experimental data, the simulation results can be used to identify preferential adsorption sites, which are located close to the organic linkers. Topological mapping of the potential-energy surfaces makes it possible to relate the preferential adsorption sites, Henry constant, and isosteric heats of adsorption at zero coverage to the nature of the host-guest interactions and the chemical nature of the organic linker. The role played by the topology of the solid and the organic linkers, instead of the metal sites, upon gas adsorption on zeolite like metal-organic frameworks is discussed. PMID- 19998435 TI - High molar extinction coefficient organic sensitizers for efficient dye sensitized solar cells. AB - We have designed and synthesized highly efficient organic sensitizers with a planar thienothiophene-vinylene-thienothiophene linker. Under standard global AM 1.5 solar conditions, the JK-113-sensitized cell gave a short circuit photocurrent density (J(sc)) of 17.61 mA cm(-2), an open-circuit voltage (V(oc)) of 0.71 V, and a fill factor (FF) of 72%, corresponding to an overall conversion efficiency (eta) of 9.1%. The incident monochromatic photo-to-current conversion efficiency (IPCE) of JK-113 exceeds 80% over the spectral region from 400 to 640 nm, reaching its maximum of 93% at 475 nm. The band tails off toward 770 nm, contributing to the broad spectral light harvesting. Solar-cell devices based on the sensitizer JK-113 in conjunction with a volatile electrolyte and a solvent free ionic liquid electrolyte gave high conversion efficiencies of 9.1% and 7.9%, respectively. The JK-113-based solar cell fabricated using a solvent-free ionic liquid electrolyte showed excellent stability under light soaking at 60 degrees C for 1000 h. PMID- 19998436 TI - Lanthanide(III) bis(phthalocyaninato)-[60]fullerene dyads: synthesis, characterization, and photophysical properties. AB - A novel series of double-decker lanthanide(III) bis(phthalocyaninato)-C(60) dyads [Ln(III)(Pc)(Pc')]-C(60) (M=Sm, Eu, Lu; Pc=phthalocyanine) (1 a-c) have been synthesized from unsymmetrically functionalized heteroleptic sandwich complexes [Ln(III)(Pc)(Pc')] (Ln=Sm, Eu, Lu) 3 a-c and fulleropyrrolidine carboxylic acid 2. The sandwich complexes 3 a-c were obtained by means of a stepwise procedure from unsymmetrically substituted free-base phthalocyanine 5, which was first transformed into the monophthalocyaninato intermediate [Ln(III)(acac)(Pc)] and further reacted with 1,2-dicyanobenzene in the presence of 1,8 diazabicyclo[5.4.0]undec-7-ene (DBU). (1)H NMR spectra of the bis(phthalocyaninato) complexes 3 a-c and dyads 1 a-c were obtained by adding hydrazine hydrate to solutions of the complexes in [D(7)]DMF, a treatment that converts the free radical double-deckers into the protonated species, that is, [Ln(III)(Pc)(Pc')H] and [Ln(III)(Pc)(Pc')H]-C(60). The electronic absorption spectra of 3 a-c and 1 a-c in THF exhibit typical transitions of free-radical sandwich complexes. In the case of dyads 1 a-c, the spectra display the absorption bands of both constituents, but no evidence of ground-state interactions could be appreciated. When the UV/Vis spectra of 3 a-c and 1 a-c were recorded in DMF, typical features of the reduced forms were observed. Cyclic voltammetry studies for 3 a-c and 1 a-c were performed in THF. The electrochemical behavior of dyads 1 a-c is almost the exact sum of the behavior of the components, namely the double-decker [Ln(III)(Pc)(Pc')] and the C(60) fullerene, thus confirming the lack of ground-state interactions between the electroactive units. Photophysical studies on dyads 1 a-c indicate that only after irradiation at 387 nm, which excites both C(60) and [Ln(III)(Pc)(Pc')] components, a photoinduced electron transfer from the [Ln(III)(Pc)(Pc')] to C(60) occurs. PMID- 19998437 TI - A micellar multitasking device: sensing pH windows and gauging the lipophilicity of drugs with fluorescent signals. AB - A multitasking fluorescent device can be obtained by forming micelles of Triton X 100, containing a lipophilic macrocyclic Cu(2+) complex and the coordinating fluorophore Coumarin 343 (C343), which features a COOH moiety. At low pH the two micellised components do not interact, and the fluorescence of Courmarin 343 (C343) is intense. At intermediate pH, C343 is deprotonated and coordinates to the Cu(2+) centre in its apical position, with fluorescence quenching. At higher pH the deprotonated C343 is displaced from Cu(2+) by the formation of an OH(-) complex, and the fluorescence is revived. This allows the system to carry out its first task as it behaves as an "on-off-on" fluorescent sensor for pH windows. The "off" part of the window ranges from pH 6 to 8. In this interval, in which the carboxylate form of C343 is apically coordinated to the Cu(2+) complex inside micelles, the device carries out its second task, that is, it behaves as a gauge for lipophilicity. For pHs between 6 and 8, molecules containing a COOH group are in their COO(-) form and distribute between bulk water and micelles proportionally to their lipophilicity. Upon entering the micelle, their COO(-) moiety competes for coordination with C343, displacing it from the Cu(2+) centre, and this results in fluorescence revival, the intensity of which is also proportional to the lipophilicity of the examined molecule. We have chosen the physiological pH value (7.4) as the working pH, and we have examined the lipophilicity of fatty acids and of the widely used family of non-steroidal anti inflammatory drugs (NSAIDs). The device successfully measures their lipophilicity, expressing it with an "off-on" type fluorescent signal, as demonstrated by the correlation of the fluorescence increase with the logarithmic water/octanol partition coefficient (log P) and with the difference between the pK(a) observed in micelles and that measured in water for NSAIDs. PMID- 19998438 TI - Structural diversity in the self-assembly of pseudopeptidic macrocycles. AB - The self-assembling abilities of several pseudopeptidic macrocycles have been thoroughly studied both in the solid (SEM, TEM, FTIR) and in solution (NMR, UV, CD, FTIR) states. Detailed microscopy revealed large differences in the morphology of the self-assembling micro/nanostructures depending on the macrocyclic chemical structures. Self-assembly was triggered by the presence of additional methylene groups or by changing from para to meta geometry of the aromatic phenylene backbone moiety. More interestingly, the nature of the side chain also plays a fundamental role in some of the obtained nanostructures, thus producing structures from long fibers to hollow spheres. These nanostructures were obtained in different solvents and on different surfaces, thus implying that the chemical information for the self-assembly is contained in the molecular structure. Dilution NMR studies (chemical shift and self-diffusion rates) suggest the formation of incipient aggregates in solution by a combination of hydrogen bonding and pi-pi interactions, thus implicating amide and aryl groups, respectively. Electronic spectroscopy further supports the pi-pi interactions because the compounds that lead to fibers show large hypochromic shifts in the UV spectra. Moreover, the fiber-forming macrocycles also showed a more intense CD signature. The hydrogen-bonding interactions within the nanostructures were also characterized by attenuated total-reflectance FTIR spectroscopy, which allowed us to monitor the complete transition from the solution to the dried nanostructure. Overall, we concluded that the self-assembly of this family of pseudopeptidic macrocycles is dictated by a synergic action of hydrogen-bonding and pi-pi interactions. The feasibility and geometrical disposition of these interactions finally render a hierarchical organization, which has been rationalized with a proposal of a model. The understanding of the process at the molecular level has allowed us to prepare hybrid soft materials. PMID- 19998439 TI - New insights into the formation and reactivity of molecular organostannonic acids. AB - The controlled base hydrolysis of 2,6-Mes(2)C(6)H(3)SnCl(3) (1; Mes=mesityl) provided 2,6-Mes(2)C(6)H(3)Sn(OH)Cl(2).H(2)O (2) and the trinuclear organostannonic acid trans-[2,6-Mes(2)C(6)H(3)Sn(O)OH](3) (3), respectively. In moist C(6)D(6), 3 reversibly reacts with water to give the monomeric organostannonic acid 2,6-Mes(2)C(6)H(3)Sn(OH)(3) (3 a). The reaction of 3 with (tBu(2)SnO)(3), Ph(2)PO(2)H, and NaH, gives rise to the multinuclear hypercoordinated organostannoxane clusters [tBu(2)Sn(OH)OSnR(OH)(2)OC(OSntBu(2)OH)(2)(O)SnR(OH)(H(2)O)](2) (5), [RSn(OH)(2)(O(2)PPh(2))](2) (6), and Na(3)(RSn)(4)O(6)(OH)(3) (7), respectively (R=2,6-Mes(2)C(6)H(3)). The characterization of the new compounds is achieved by multinuclear NMR spectroscopy and electrospray mass spectrometry in solution and (119)Sn MAS NMR spectroscopy, IR spectroscopy, and X-ray crystallography in the solid-state. PMID- 19998440 TI - Facile synthesis of highly stable tetraazaheptacene and tetraazaoctacene dyes. PMID- 19998441 TI - Levels of oxidative damage and lipid peroxidation in thyroid neoplasia. AB - BACKGROUND: This study assessed the presence of oxidative damage and lipid peroxidation in thyroid neoplasia. METHODS: Using tissue microarrays and immunohistochemistry, we assessed levels of DNA damage (8-oxo-dG) and lipid peroxidation (4-HNE) in 71 follicular thyroid adenoma (FTA), 45 papillary thyroid carcinoma (PTC), and 17 follicular thyroid carcinoma (FTC) and matched normal thyroid tissue. RESULTS: Cytoplasmic 8-oxo-dG and 4-HNE expression was significantly higher in FTA, FTC, and PTC tissue compared to matched normal tissue (all p values < .001). Similarly, elevated nuclear levels of 8-oxo-dG were seen in all in FTA, FTC, and PTC tissue compared to matched normal (p values < .07, < .001, < .001, respectively). In contrast, a higher level of 4-HNE expression was detected in normal thyroid tissue compared with matched tumor tissue (p < .001 for all groups). Comparing all 3 groups, 4-HNE levels were higher than 8-oxo-dG levels (p < .001 for all groups) except that cytoplasmic levels of 8-oxo-dG were higher than 4-HNE in all (p < .001). These results were independent of proliferation status. CONCLUSION: High levels of DNA damage and lipid peroxidation in benign and malignant thyroid neoplasia indicates this damage is an early event that may influence disease progression. PMID- 19998442 TI - Feasibility and surgical approach of transaxillary robotic thyroidectomy without CO(2) insufflation. AB - BACKGROUND: Our objective was to evaluate the anatomic basis for robotic-assisted transaxillary thyroidectomy and to determine its feasibility in a prospective clinical trial. METHODS AND RESULTS: Using the da Vinci Surgical Robotic System, we performed 5 cadaveric dissections, via transaxillary approach without gas insufflation. Once the safety and feasibility of this approach had been demonstrated in cadavers, it was utilized to perform a thyroid lobectomy in a patient. The da Vinci system provided excellent visualization of the recurrent and superior laryngeal nerves, parathyroid glands, and paratracheal lymphatics. After the 5 cadaver dissections, the procedure time diminished from >90 minutes to <30 minutes. CONCLUSION: Robotic-assisted transaxillary thyroidectomy is feasible with proper instrumentation and an understanding of the surgical anatomy. Based on this preclinical laboratory study and our experience in 1 patient, further evaluation of this approach in the setting of a prospective clinical trial is warranted to determine standardized criteria identifying patients who would benefit from this approach. PMID- 19998443 TI - Cytogenetics of long-term survivors of ETV6-RUNX1 fusion positive acute lymphoblastic leukemia. AB - This study describes the cytogenetics of 33 children with ETV6-RUNX1 positive acute lymphoblastic leukemia (ALL) who had been in continuous complete remission for a minimum of 8.8 years [median event-free survival (EFS) 10.9 years]. The results were compared with a published series of 16 fusion positive patients treated on the same childhood ALL trial, who had relapsed (median EFS, 2.3 years). Interphase fluorescence in situ hybridization (FISH) at diagnosis showed deletion of the second ETV6 signal from all fusion positive cells in 45% of the long-term survivors but in none of the relapsed patients, whereas patients with mixed populations with retained or lost second signals were more frequent among those who had relapsed (69%) than the long-term survivors (21%). Interphase populations with two fusion signals in 18% of the long-term survivors and 31% of relapsed patients were smaller in the long-term survivors (median, 4% of total cells) than in the relapsed patients (median, 84%). The additional copy of chromosome 21 in 30% of long-term survivors and in 69% of relapsed patients was a derived chromosome 21 in 20% and 55% of patients, respectively. Metaphase FISH for 26 long-term survivors and 15 relapsed patients revealed complex karyotypes in both groups. Variant translocations involved different chromosome arms between the long-term survivors and relapsed patients. It appears that the two groups have some distinguishing cytogenetic features at the time of diagnosis, which may provide pointers to relapse that are worthy of more detailed study. PMID- 19998444 TI - Telomere shortening and chromosomal instability in myelodysplastic syndromes. AB - Telomere shortening and chromosomal instability are believed to play an important role in the development of myeloid neoplasia. So far, published data are only available on the average telomere length in myelodysplastic syndromes (MDS) and acute myeloid leukemia (AML), but not on the telomere length of individual chromosomes. We used a new technique, telomere/centromere-fluorescence in situ hybridization (T/C-FISH), which combines fluorescence R-banding and FISH using a probe against the telomere repeats to measure the telomere length of each chromosome arm in 78 patients with MDS. In line with the previous results, patients with MDS showed significantly shorter telomeres than those of healthy controls. Telomere lengths did not differ significantly between distinct morphological subtypes of MDS. However, there was a significant difference in telomere length between patients with an isolated monosomy 7 and patients with a normal karyotype (P < 0.05). Notably, patients with an isolated monosomy 7 showed significantly longer telomeres than patients with a normal karyotype in many chromosome arms, among them 7p and 7q. Neo-telomeres were found in two patients with a complex karyotype, in one case at the fusion site of a dic(14;20). Normal and aberrant metaphases of the same patient did not differ in telomere length, thus indicating to telomere shortening as a basic mechanism affecting all hematopoietic cells in patients with MDS. In some MDS subtypes, like MDS with isolated monosomy 7, telomeres may be stabilized and even increase in length because of the activation of telomerase or alternative mechanisms. PMID- 19998445 TI - A multimethod study of needs for physician assessment: implications for education and regulation. AB - INTRODUCTION: Deficiencies in physician competence play an important role in medical errors and poor-quality health care. National trends toward implementation of continuous assessment of physicians hold potential for significant impact on patient care because minor deficiencies can be identified before patient safety is threatened. However, the availability of assessment methods and the quality of existing tools vary, and a better understanding of the types of deficiencies seen in physicians is required to prioritize the development and enhancement of assessment and remediation methods. METHODS: Surveys of physicians and licensing authorities and analysis of the Federation of State Medical Boards (FSMB) Board Action Data Bank were used to collect information describing the nature and types of problems seen in practicing physicians. Focus groups, depth interviews with key professional stakeholders, and state medical board site visits provided additional information about deficiencies in physician competence. RESULTS: Quantitative and qualitative analyses identified (1) communication skills as a priority target for assessment approaches that also should focus on professional behaviors, knowledge, clinical judgment, and health-care quality; and (2) differences between regulatory approaches of licensing and certifying bodies contribute to a culture that limits effective self-assessment and continuous quality improvement. System problems impacting physician performance emerged as an important theme in the qualitative analysis. DISCUSSION: Considering alternative perspectives from the regulatory, education, and practice communities helps to define assessment priorities for physicians, facilitating development of a coherent and defensible approach to assessment and continuing professional development that promises to provide a more comprehensive solution to problems of health-care quality in the United States. PMID- 19998446 TI - Interrater reliability to assure valid content in peer review of CME-accredited presentations. AB - INTRODUCTION: The Accreditation Council for Continuing Medical Education (ACCME) provides guidelines for continuing medical education (CME) materials to mitigate problems in the independence or validity of content in certified activities; however, the process of peer review of materials appears largely unstudied and the reproducibility of peer-review audits for ACCME accreditation and designation of American Medical Association Category 1 Credit(TM) is unknown. METHODS: Categories of presentation defects were constructed from discussions of the CME committee of the American Epilepsy Society: (1) insufficient citation, (2) poor formatting, (3) nonacknowledgment of non-FDA-approved use, (4) misapplied data, (5) 1-sided data, (6) self- or institutional promotion, (7) conflict of interest/commercial bias, (8) other, or (9) no defect. A PowerPoint lecture (n = 29 slides) suitable for presentation to general neurologists was purposefully created with the above defects. A multirater, multilevel kappa statistic was determined from the number and category of defects. RESULTS: Of 14 reviewers, 12 returned completed surveys (86%) identifying a mean +/- standard deviation 1.6 +/ 1.1 defects/slide. The interrater kappa equaled 0.115 (poor reliability) for number of defects/slides. No individual categories achieved kappa > 0.38. DISCUSSION: Interrater reliability on the rating of durable materials used in subspecialty CME was poor. Guidelines for CME appropriate content are too subjective to be applied reliably by raters knowledgeable in their specialty field but relatively untrained in the specifics of CME requirements. The process of peer review of CME materials would be aided by education of physicians on validation of materials appropriate for CME. PMID- 19998448 TI - The use of medical information in Nigeria: the influence of gender and status. AB - INTRODUCTION: Medical doctors are important members of the health-care team, and to discharge their duties credibly, they need adequate and quick information. A search of the literature yielded a handful of studies on the provision and organization of medical information with little emphasis on the influence of personal variables on information utilization. We report the influence of gender and status on reasons for searching medical information by doctors in Nigeria. METHODS: A pretested questionnaire was given to all 720 doctors in active practice working in the 5 Teaching Hospitals of southeastern Nigeria. The returned questionnaires were analyzed by analysis of variance (ANOVA). RESULTS: Six hundred fourteen of 720 (85.3%) doctors responded to the questionnaire; 116 (18.9%) of the respondents were females. It was found that among the male resident doctors the most common reason for searching medical information was for examination preparation, which had 356 (89.9%) positive respondents, followed by career development, 340 (85.9%). For the female residents, updating knowledge was the most common reason, with 90 (91.8%) positive respondents followed by examination preparation, 87 (88.8%). Updating knowledge was the most common reason for both the male and female consultants, followed by research for teaching and publication. DISCUSSION: The male resident doctors were more concerned with passing their professional examinations, and that was their main reason for searching medical information. The other groups searched information mostly to update their knowledge. Better medical practice was not an important reason to any of the groups. This attitude should be changed through continuing medical education (CME), which refers to a specific form of continuing education that helps those in the medical field maintain competence and learn about new and developing areas of their field. PMID- 19998447 TI - Characteristics that predict physician participation in a Web-based CME activity: the MI-Plus study. AB - INTRODUCTION: Physician use of the Internet for practice improvement has increased dramatically over the last decade, but research shows that many physicians choose not to participate. The current study investigated the association of specific physician characteristics with enrollment rates and intensity of participation in a specific Internet-delivered educational intervention to improve care to post-myocardial infarction (MI) patients. METHODS: Primary-care physicians were recruited for participation in a randomized controlled trial designed to compare effectiveness of an intervention Web site versus a control Web site in the management of adult chronic disease. Physicians were informed that the intervention focused on ambulatory post-myocardial infarction patients. Physician characteristics were obtained from a commercial vendor with data merged from the American Medical Association and Alabama State Licensing Board. Enrollment and Web use were tracked electronically. RESULTS: Out of a sample of 1337 eligible physicians, 177 (13.2%) enrolled in the study. Enrollment was higher for physicians with more post-MI patients (> or = 20 vs < 20 patients, 15.3% vs 9.3%, P = .002) and for those practicing in rural compared to urban areas (16.3% vs 12.1%, P = .046). Intensity of use of the Internet courses after initial enrollment was not predicted by physician characteristics in the current sample. DISCUSSION: Physicians with more post-MI patients and rural practice location were found to predict enrollment in an Internet-delivered continuing medical education (CME) intervention designed to improve care for post MI patients. These factors predicted program interest but not program use. More research is needed to replicate these findings to investigate variables that determine physician engagement in Internet CME. PMID- 19998449 TI - Polymorphisms in the transforming growth factor beta 1 pathway in relation to colorectal cancer progression. AB - Transforming growth factor beta1 (TGFB1) acts as a growth inhibitor of normal colonic epithelial cells, however, as a tumor promoter of colorectal cancer (CRC) cells. To explore the association between genetic polymorphisms in the TGFB1 pathway and CRC susceptibility and clinical outcome, we carried out a case control study on a Swedish population of 308 CRC cases and 585 age- and gender matched controls. The cases were sampled prospectively and had up to 16 years follow-up, making the study material particularly suitable for survival analysis. On the basis of their reported or predicted functional effect, nine single nucleotide polymorphisms (TGFB1: Leu10Pro; TGFBR1: 9A/6A and IVS7G+24A; FURIN: C 229T; THBS1: T+42C; LTBP1L: C-256G; LTBP4: T-893G and Thr750Ala; BAMBI: T-779A) were selected for genotyping. We evaluated the associations between genotypes and CRC and Dukes' stage. Survival probabilities were compared between different subgroups. The observed statistically significant associations included a decreased CRC risk for TGFBR1 IVS7G+24A minor allele carriers (odds ratio (OR): 0.72, 95% confidence interval (CI): 0.53-0.97), less aggressive tumors with Dukes' stage A+B for carriers of LTBP4 Thr750Ala and BAMBI T-779A minor alleles (OR: 0.58, 95%CI: 0.36-0.93 and OR: 0.51, 95%CI: 0.29-0.89, respectively) and worse survival for FURIN C-229T heterozygotes (hazard ratio: 1.63, 95%CI: 1.08 2.46). As this is the first study about the influence of the polymorphisms in the TGFB1 pathway on CRC progression, further studies in large independent cohorts are warranted. PMID- 19998450 TI - Teaching and evaluating point of care learning with an Internet-based clinical question portfolio. AB - INTRODUCTION: Diplomates in the American Board of Internal Medicine (ABIM) Maintenance of Certification (MOC) program satisfy the self-evaluation of medical knowledge requirement by completing open-book multiple-choice exams. However, this method remains unlikely to affect practice change and often covers content areas not relevant to diplomates' practices. We developed and evaluated an Internet-based point of care (POC) learning portfolio to serve as an alternative. METHODS: Participants enter information about their clinical questions, including characteristics, information pursuit, application, and practice change. After documenting 20 questions, they reflect upon a summary report and write commitment to-change statements about their learning strategies. They can link to help screens and medical information resources. We report on the beta test evaluation of the module, completed by 23 internists and 4 internal medicine residents. RESULTS: Participants found the instructions clear and navigated the module without difficulty. The majority preferred the POC portfolio to multiple-choice examinations, citing greater relevance to their practice, guidance in expanding their palette of information resources, opportunity to reflect on their learning needs, and "credit" for self-directed learning related to their patients. Participants entered a total of 543 clinical questions, of which 250 (46%) resulted in a planned practice change. After completing the module, 14 of 27 (52%) participants committed to at least 1 change in their POC learning strategies. DISCUSSION: Internists found the portfolio valuable, preferred it to multiple-choice examinations, often changed their practice after pursuing clinical questions, and productively reflected on their learning strategies. The ABIM will offer this portfolio as an elective option in MOC. PMID- 19998451 TI - Self-directed learning needs, patterns, and outcomes among general surgeons. AB - INTRODUCTION: To explore the relationship between self-directed learning (SDL) needs, patterns, barriers, and outcomes among nonacademic general surgeons. METHODS: Participants dictated details of SDL episodes associated with cancer patient management from October 2007 to March 2008. Transcripts were coded thematically. Frequencies were calculated for elements of each SDL stage. Statistical significance among subgroups was established with the use of the Pearson chi-square test, adjusted for clustering by surgeon. Participants were interviewed by telephone, and transcripts were analyzed by qualitative methods. RESULTS: Of 21 consenting surgeons, 15 submitted 115 cases, and 108 were analyzed. Most involved breast (40.7%), colon (18.5%), or rectal cancer (13.0%); 2 or more clinical tasks (41.7%); and 2 or more questions (89.8%). Information was sought from the Internet (48.1%), colleagues (24.2%), or both (6.8%). Information was partially, or not relevant for 21.3% of cases. Evidence was new for 66.7%, and confirmed knowledge for 10.7% of cases. Learning helped surgeons formulate new (34.2%), or confirm original (16.5%) management plans, or determine that referral was appropriate (39.2%). Use of codified sources was associated with information retrieval (P < .05), and identifying new evidence leading to a change in management from that initially proposed (P < or = .001). DISCUSSION: Numerous individual and systemic barriers may prevent practicing physicians from undertaking SDL, but provision of structured guidance prompted SDL and resulted in several beneficial outcomes. Further research is needed to validate these findings, and investigate who should support SDL, and how. PMID- 19998452 TI - Information seeking in the digital age - Why closing knowledge gaps is not education and why the difference matters. PMID- 19998454 TI - Can fecal calprotectin or lactoferrin identify postoperative recurrence in Crohn's disease? PMID- 19998453 TI - Physician-pharmacist collaborative care for dyslipidemia patients: knowledge and skills of community pharmacists. AB - INTRODUCTION: In a physician-pharmacist collaborative-care (PPCC) intervention, community pharmacists were responsible for initiating lipid-lowering pharmacotherapy and adjusting the medication dosage. They attended a 1-day interactive workshop supported by a treatment protocol and clinical and communication tools. Afterwards, changes in pharmacists' knowledge, their skills, and their satisfaction with the workshop were evaluated. METHODS: In a descriptive study nested in a clinical trial, pharmacists assigned to the PPCC intervention (n = 58) completed a knowledge questionnaire before and after the workshop. Their theoretical skills were evaluated with the use of a vignette approach (n = 58) after the workshop and their practical skills were assessed by direct observation with study patients (n = 28). RESULTS: The mean (SD) overall knowledge score was 45.8% (12.1%) before the workshop; it increased significantly to 89.3% (8.3%) afterwards (mean difference: 43.5%; 95% CI: 40.3%-46.7%). All the pharmacists had an overall theoretical-skill score of at least 80%, the minimum required to apply the PPCC in the trial. From 92.9% to 100% of the pharmacists' interventions with study patients complied with the treatment protocol. DISCUSSION: In primary care, a short continuing-education program based on a specific treatment protocol and clinical tools is necessary and probably sufficient to prepare pharmacists to provide advanced pharmaceutical care. PMID- 19998455 TI - Interleukin 21 expression is increased in rectal biopsies from patients with ulcerative colitis. PMID- 19998456 TI - CD24 is upregulated in inflammatory bowel disease and stimulates cell motility and colony formation. AB - BACKGROUND: We investigated whether CD24 (reportedly a stem cell marker and adhesion molecule) was expressed in regenerative mucosa in inflammatory bowel disease (IBD) and whether it could be functionally relevant. METHODS: CD24 expression was examined in 10 cases of IBD and the relationship of CD24 with Wnt signaling was tested using dominant negative (DN)-TCF4 expression. For functional evaluation, CD24 was 1) cloned and forcibly expressed in HCT116 (which expresses very low levels of CD24) and 2) knocked-down by RNA interference in HT29 (which expresses high levels of CD24). The effect of altered CD24 expression on proliferation/apoptosis, staurosporine-induced apoptosis, colony formation in soft agar, migration, and invasion was examined. RESULTS: CD24 was not expressed in normal tissue, while 10/10 cases of IBD showed CD24 upregulation. Inhibition of Wnt signaling with DN-TCF4 caused CD24 downregulation. Forced expression of CD24 did not influence cell proliferation, apoptosis, or staurosporine-induced apoptosis but it did significantly enhance colony forming efficiency (P < 0.01). Furthermore, there was increased transwell migration (P < 0.001) and invasion (P < 0.03) and there was increased cell migration in wounding assays. Conversely, knockdown of CD24 reduced transwell migration (P < 0.01) and invasion (P < 0.01) and reduced cell motility in wounding assays. CD24 knockdown did not influence proliferation, apoptosis resistance, or staurosporine-induced apoptosis. CONCLUSIONS: This is the first study to report upregulation of CD24 in regenerating tissue in IBD. This may be regulated by Wnt signaling and can confer enhanced colony forming ability and enhanced cell motility-features that may be important in tissue healing in the colon. PMID- 19998457 TI - Ocular manifestations in a community-based cohort of patients with inflammatory bowel disease. AB - BACKGROUND: Ophthalmologic diseases in patients with inflammatory bowel disease (IBD) have been reported with varying frequency, mostly from tertiary referral centers. The aim was to describe the spectrum of ophthalmologic conditions in patients with IBD in a community setting and to compare it with a control non-IBD cohort. METHODS: A prospective cohort of patients with Crohn's disease (CD), ulcerative colitis (UC), and non-IBD controls underwent evaluation by an ophthalmologist, including visual acuity, slit-lamp exam, and assessment of lacrimal output. RESULTS: The ophthalmologic exam was completed in 112 subjects; 48 with CD, 40 with UC, and 24 controls. Active intestinal disease was present in 52/88 (59%) of the IBD patients, and 79/88 (89%) were taking 5-aminosalicylates (5-ASAs). The IBD and control populations had similar age and gender profiles. Patients with IBD were more likely to report ocular symptoms (odds ratio [OR] 5.6, 95% confidence interval [CI] 1.5-20), particularly dry eyes (OR 5.3, 95% CI 1.4-19), than the control population. On objective exam, 42% of IBD patients had evidence of dry eyes. In a univariate analysis, 5-ASA use was associated with an increased risk of ocular symptoms (OR 7.4, 95% CI 0.9-64), and 5-ASA use >3 g per day was associated with an increased odds ratio of dry eyes (OR 15, 95% CI 1.9 122). Active disease was not associated with eye symptoms or dry eyes. Other eye conditions such as episcleritis, uveitis, or cataracts were infrequent in this cohort. CONCLUSIONS: Patients with IBD in the community frequently have dry eyes. This is associated with 5-ASA use, particularly doses >3 g per day. Whether this is a surrogate marker of disease severity is unclear. PMID- 19998458 TI - Nutriose, a prebiotic low-digestible carbohydrate, stimulates gut mucosal immunity and prevents TNBS-induced colitis in piglets. AB - BACKGROUND: We investigated a prebiotic low-digestible carbohydrate (LDC) as a possible food ingredient to stimulate bowel functions in the treatment of inflammatory bowel disease. The study aimed to assess a fermentable dextrin fiber (Nutriose) and its relationship to the immune management of the disease and the microbiota profile in colitis-bearing piglets. METHODS: In a randomized placebo controlled parallel blind preclinical study, 32 male piglets were fed LDC (4% Nutriose) or dextrose placebo for 44 days before being challenged with trinitrobenzene sulfonic acid (TNBS) to induce colitis. We followed the microbiota profile using real-time polymerase chain reaction (PCR) targeted to 9 bacterial genera. Secretory IgA was evaluated by enzyme-linked immunosorbent assay (ELISA). Inflammatory protein profiles were monitored in blood and colonic tissues. Both histological scoring of biopsy samples and live endoscopic scoring were used to measure colitis development. RESULTS: Prior and continuing LDC supplementation alleviated the symptoms of colitis (body weight loss, bloody stools) induced by a TNBS challenge. This effect was associated with an improvement in endoscopic and histological scores. LDC was shown to selectively downregulate some of the proinflammatory factors and their concomitant pyretic events and to stimulate the Th2-related immune pathway (IL-10 and s-IgA). CONCLUSIONS: At the dose tested, LDC is a well-tolerated prebiotic agent able to not only stimulate butyrogenic bacteria strains and reduce intestinal transit disorders and energy intake, but also to prevent chronic inflammatory intestinal injuries. PMID- 19998459 TI - Population-based Birth Defects Surveillance data from selected states, 2002-2006. PMID- 19998460 TI - Quality and publication success of abstracts of randomized clinical trials in inflammatory bowel disease presented at Digestive Disease Week. AB - BACKGROUND: The incorporation of abstracts from scientific meetings into systematic reviews and practice guidelines may reduce publication bias and delays in implementing therapeutic interventions. METHODS: All abstracts of Phase III randomized controlled trials in inflammatory bowel disease accepted at Digestive Disease Week (1998-2003) were identified. MedLine, PubMed (1997-current), EMBASE, and Google Scholar were searched for subsequent full publications. Characteristics of methodology and outcomes of the abstracts and articles were analyzed. RESULTS: The 5-year cumulative publication rate of the 82 eligible abstracts was 78%. Abstracts that presented negative results were less likely to be published than those with positive findings, particularly after the first 2 years (hazard ratio 6.45; 95% confidence interval [CI]: 2.22-18.7) with 5-year cumulative publication rates of (50% versus 91%, respectively, P < 0.001). The median time to publication was longer for negative than positive abstracts (58 versus 26 months, P < 0.001). Abstracts selected for oral presentation were more likely to be published than poster presentations (89% versus 69%; P = 0.03). A change in primary outcome results was observed in 28% (n = 18) of abstracts compared to that in the final publication, and 6% (n = 4) had a statistically significant change resulting in a change of study conclusions. CONCLUSIONS: Our findings suggest that the use of abstract data would enable detection and mitigation of publication bias. Improving the uniformity and quality of abstract reporting of randomized clinical trials at scientific meetings may facilitate their incorporation in practice guidelines and systematic reviews. PMID- 19998461 TI - Asymmetric endoscopic inflammation of the ileal pouch: a sign of ischemic pouchitis? AB - BACKGROUND: Pouchitis is associated with dysbiosis and dysregulated mucosal immunity, although secondary pouchitis with special etiologic factors, such as ischemia, can occur. The aim was to describe a disease phenotype of the ileal pouch with an endoscopic appearance suggestive of ischemia. METHODS: We identified consecutive patients with endoscopic asymmetric inflammation of the pouch (inflammation of side of the pouch with a completely normal other limb of the pouch one limb and a sharp demarcation along the staple suture line). Patients with Crohn's disease (CD) of the pouch or antibiotic-responsive pouchitis, matched for duration of the pouch, served as controls. Histology slides of mucosal biopsies were re-reviewed independently by 2 blinded gastrointestinal pathologists. Demographic, clinical, endoscopic, histologic, and imaging characteristics were compared between the groups. RESULTS: Ten patients with "ischemic" pouchitis, 15 with CD of the pouch, and 15 with antibiotic responsive pouchitis were studied. Pyloric gland metaplasia was observed only in the groups with CD of the pouch (23.1%) or antibiotic-responsive pouchitis (13.3%). Of patients with "ischemic" pouchitis, 80% had extracellular hemosiderin or hematoidin deposits (versus 30.8% those with CD of the pouch and 13.3% of those with pouchitis, P = 0.003). The majority of patients (80%) with "ischemic" pouchitis did not respond to conventional antibiotic therapy. It appeared that subsequent abdominal surgeries after pouch construction and a history of postoperative portal vein thrombi were associated with "ischemic" pouchitis. CONCLUSIONS: Endoscopic asymmetric inflammation of the pouch may represent an ischemia-associated pouchitis with characteristic clinical, radiographic, and histologic features. Its hemodynamic, cellular, and molecular basis of mechanism warrants further study. PMID- 19998463 TI - Genetic Drift. Word smithing in medical genetics. AB - Certain terms used in medical genetics and more often in other medical fields are in need of clarification. The terms cited are frequently misunderstood, mispronounced, and/or misspelled. The discussion includes two Latin-derived terms (genua valga and calvaria), one Greek-derived term (apoptosis), one gene-derived term (RUNX), one syndromic eponym (Kartagener syndrome), and three vascular eponyms (Klippel-Trenaunay syndrome, Parkes Weber syndrome, and Kasabach-Merritt phenomenon). PMID- 19998462 TI - Health-related quality of life of youth with inflammatory bowel disease: a comparison with published data using the PedsQL 4.0 generic core scales. AB - BACKGROUND: This study compared youth and parent-proxy reports of health-related quality of life (HRQoL) among youth with inflammatory bowel disease (IBD) to published comparison group data and examined concordance between youth and parent proxy reports of HRQoL. METHODS: One hundred thirty-six youth and parent-proxy reports on the PedsQL 4.0 Generic Core Scales were compared to published data from chronically ill, acutely ill, and healthy comparison groups using independent samples t-tests. Reporter agreement was examined using paired samples t-tests and intraclass correlations (ICCs). RESULTS: Youth with IBD reported lower psychosocial functioning than the healthy comparison group, higher physical and social functioning than the chronically ill group, and lower school functioning than all published comparison groups. Parent-proxy reports of youth HRQoL were higher than the chronically ill group, but lower than the healthy group on all scales except psychosocial functioning. Youth with active IBD reported lower physical health domain scores than youth with inactive disease. Concordance between youth and parent-proxy reports was moderate, with the lowest agreement in school and social functioning. CONCLUSIONS: Youth with IBD and their parents rate HRQoL as lower than healthy youth but do not perceive the impact of IBD to be as limiting as in other chronic conditions. Youth report suggests that IBD may be particularly detrimental to HRQoL in the school functioning domain. Moderate agreement between parent and youth reports substantiates continued use of multiple informants in studies of pediatric HRQoL. PMID- 19998464 TI - Effect of recombinant galectin-1 on the growth of immortal rat chondrocyte on chitosan-coated PLGA scaffold. AB - The effect of galectin-1 (GAL1) on the growth of immortal rat chondrocyte (IRC) on chitosan-modified PLGA scaffold is investigated. The experimental results showed that water absorption ratio of chitosan-modified PLGA scaffold was 70% higher than that of PLGA alone after immersion in ddH(2)O for 2 weeks, indicating that chitosan-modification significantly enhances the hydrophilicity of PLGA. The experimental results also showed that GALl efficiently and spontaneously coats the chitosan-PLGA scaffold surface to promote adhesion and growth of immortal rat chondrocyte (IRC). To investigate the effect of endogenous GAL1, the full-length GAL1 cDNAs were cloned and constructed into pcDNA3.1 vectors to generate a plasmid expressed in IRC (IRC-GAL1). The results showed that IRC-GAL1 growth was significantly higher than that of IRC on chitosan-PLGA scaffold. The GAL1 potentiated IRC growth on chitosan-PLGA scaffold was dose-dependently inhibited by TDG (specific inhibitor of GAL1 binding). These results strongly suggest that GAL1 is critical for enhancing IRC cell adhesion and growth on chitosan-PLGA scaffold. Moreover, GAL1-coating or expression tends to promote IRC cell-cell aggregation on chitosan-PLGA scaffold and significantly enhances IRC migration. These results suggest that GAL1 probably could induce tissue differentiation and facilitates cartilage reconstruction. In conclusion, the experimental results suggest that both GAL1 and chitosan are important for enhancing IRC cell adhesion and growth on PLGA scaffold, and GAL1 is a potential biomaterial for tissue engineering. PMID- 19998465 TI - Central nervous system involvement at the time of presentation in acute promyelocytic leukemia. AB - Central nervous system (CNS) involvement is rarely observed in acute promyelocytic leukemia (APML). Most cases of CNS involvement occur at relapse rather than at presentation. Because of the extremely low incidence of CNS disease, diagnostic lumbar puncture is not routinely required and prophylactic intrathecal chemotherapy is not routinely administered. Here, we describe a teenage patient with newly diagnosed APML, chloromas, and symptomatic CNS involvement confirmed by MRI and cerebrospinal fluid (CSF) findings. PMID- 19998466 TI - Red cell transfusions for patients with sickle cell disease: do we have more to learn? PMID- 19998467 TI - Spontaneous tumor lysis syndrome in a child with T-cell acute lymphoblastic leukemia. AB - We report a 5-year-old female who presented with unexplained acute renal failure (ARF) and hyperuricemia and who was subsequently diagnosed of T-cell acute lymphoblastic leukemia (ALL). Peripheral smear was initially unremarkable. She required hemodialysis. Two weeks later, peripheral smear showed 40% blasts and bone marrow demonstrated T-cell ALL. Our case was the fifth and the youngest case of ALL with spontaneous tumor lysis syndrome. However, in contrast to previous reports in ALL or acute myeloid leukemia, our patient did not have blasts noted on periphereal blood smear and her white blood cell count and serum lactate dehydrogenase level were normal on admission, a time when dialysis-dependent ARF and severe hyperuricemia were present. Occult hematologic malignancy should be considered in cases of ARF and hyperuricemia of unknown etiology even when peripheral hematologic findings are not informative. PMID- 19998468 TI - Pseudometastasis secondary to histoplasmosis infection: false-positive PET/CT findings. AB - Positron emission computed tomography (PET)/CT is a common modality used in the workup of neoplastic conditions. However, false-positive results may be produced by underlying infectious processes. We report two cases of false-positive PET/CT studies secondary to histoplasmosis infections in both a pediatric and a young adult patient. After the diagnoses, one patient was observed with no therapy and the other received a complete course of itraconazole. In both cases biopsy results were positive for fungal elements consistent with histoplasmosis. Both patients were successfully managed and have had follow-up histoplasmosis titers that reveal resolving infections. PMID- 19998469 TI - Successful mobilization, harvest and transplant of peripheral blood stem cells using AMD3100 and G-CSF following high dose craniospinal irradiation for medulloblastoma in a young child. AB - Contemporary protocols for the treatment of malignant brain tumors such as medulloblastoma (MB) in children, often involve craniospinal irradiation (CSI) at diagnosis followed by serial courses of high dose chemotherapy and autologous hematopoietic stem cell support. Patients often require several pheresis procedures in order to collect sufficient stem cells for this type of treatment, particularly if they have already had CSI. We describe the successful mobilization, collection and subsequent transplant of a 7-year-old female with medulloblastoma after recent CSI using granulocyte colony stimulating factor (G CSF) and the CXCR4 antagonist AMD3100 after a failed previous mobilization attempt using G-CSF alone. PMID- 19998470 TI - Thiotepa/topotecan/carboplatin with autologous stem cell rescue in recurrent/refractory/poor prognosis pediatric malignancies of the central nervous system. AB - BACKGROUND: Thiotepa and carboplatin are known to be active in central nervous system tumors. Topotecan potentiates the anti-cancer effects of alkylators and crosses the blood-brain barrier. We present ten patients with recurrent or progressive central nervous system malignancies treated on a myeloablative regimen using these drugs. METHODS: Treatment included: Thiotepa 300 mg/m(2) on days -8, -7, and -6; topotecan 2 mg/m(2) on days -8, -7, -6, -5, and -4; and carboplatin approximately 500 mg/m(2) (Calvert formula-area under the curve = 7) on days -5, -4, and -3. Stem cell rescue was on day 0. RESULTS: Age at study entry ranged from 2.5 to 20 years old (median age 8.7 years). Five had medulloblastoma (MB), four had high grade glioma (HGG), and one had trilateral retinoblastoma/pineoblastoma (tRB/PB). Prior treatment for all patients included surgery and chemotherapy (1-7 regimens, median 2). Nine patients received radiotherapy; one patient did not receive radiotherapy pre-study. Three patients had residual disease at the time of transplant. There were two toxic deaths. Four patients are event-free survivors at a median of 6 years (range 2.8-7.6 years) after treatment including 2/5 MB patients, 1/4 HGG patients, and the tRB/PB patient. Four of the seven patients with no evidence of disease/minimal residual disease status at the time of stem cell rescue are long-term survivors versus 1/3 with measurable disease. CONCLUSION: Thiotepa/topotecan/carboplatin may help consolidate remission of poor prognosis pediatric central nervous system tumors. Diagnosis and extent of disease prior to stem cell rescue may have an impact on outcome. PMID- 19998471 TI - Tumor vaccination for malignant brain tumors. PMID- 19998472 TI - Right foot congenital infantile fibrosarcoma treated only with chemotherapy. AB - Congenital infantile fibrosarcoma (CIF) is a rare tumor in childhood. The 5-year survival rate for CIFs is high and has been reported between 84% and 93%, but limb-amputation/disarticulation is still a major problem. We report the case of a male newborn with a mass in his right foot. X-ray and MRI revealed a mass destroying all tarsal, metatarsal, and phalangeal bones. The patient was treated only with VAC chemotherapy and is able to walk normally. PMID- 19998473 TI - Pancreatic tumors in children and adolescents: the Italian TREP project experience. AB - INTRODUCTION: Malignant pancreatic tumors are exceedingly rare in pediatric age and their clinical features and treatment usually go unappreciated by most pediatric oncologists and surgeons. METHODS: From January 2000 to July 2009, 21 patients <18 years old with pancreatic tumors were prospectively registered in the Italian cooperative TREP project dedicated to very rare pediatric tumors. RESULTS: Tumor types were 4 pancreatoblastomas, 2 pancreatic carcinomas, 3 neoplasms of the endocrine pancreas, and 12 solid pseudopapillary tumors. Three of the four patients with pancreatoblastoma had advanced disease at diagnosis and were given chemotherapy; at the time of this report, three patients were alive in first remission, while one died due to treatment toxicity. Both the cases of pancreatic carcinoma had the acinar cell subtype and successfully underwent pancreaticoduodenectomy with complete tumor resection, remaining without evidence of disease at the time of this analysis. The histological diagnoses of the three endocrine tumors were a malignant islet cell tumor, a gastrinoma, and a well differentiated tumor. All 12 patients with solid pseudopapillary tumors underwent complete tumor resection and were given no adjuvant treatment; 11 were alive in first remission, while one experienced a local and distant relapse 5 years after diagnosis. CONCLUSIONS: Surgery remains the keystone of treatment for pancreatic tumors in pediatric age as in adults. The TREP project shows that prospective cooperative studies are feasible even for such very rare tumors as these and may serve as a model for developing international cooperative schemes. PMID- 19998474 TI - UK health-care professionals' experience of on-line learning techniques: a systematic review of qualitative data. AB - INTRODUCTION: Continuing professional development and education is vital to the provision of better health services and outcomes. The aim of this study is to contribute to the evidence base by performing a systematic review of qualitative data from studies reporting health professionals' experience of e-learning. No such previous review has been published. METHOD: A systematic review of qualitative data reporting UK health professionals' experiences of the ways in which on-line learning is delivered by higher education and other relevant institutions. Evidence synthesis was performed with the use of thematic analysis grounded in the data. RESULTS: Literature searches identified 19 relevant studies. The subjects of the studies were nurses, midwives, and allied professions (8 studies), general practitioners and hospital doctors (6 studies), and a range of different health practitioners (5 studies). The majority of courses were stand-alone continuing professional development modules. Five key themes emerged from the data: peer communication, flexibility, support, knowledge validation, and course presentation and design. DISCUSSION: The effectiveness of on-line learning is mediated by the learning experience. If they are to enhance health professionals' experience of e-learning, courses need to address presentation and course design; they must be flexible, offer mechanisms for both support and rapid assessment, and develop effective and efficient means of communication, especially among the students themselves. PMID- 19998475 TI - German ambulatory care physicians' perspectives on continuing medical education - a national survey. AB - INTRODUCTION: This survey aimed to investigate German ambulatory physicians' opinions about mandatory continuing medical education (CME) and CME resources shortly before the introduction of mandatory CME in 2004. METHODS: A structured national telephone survey of general practitioners and specialists was conducted. Main outcome measures were opinions about mandatory CME, preferred CME media, and financial aspects of CME. Data analysis applied Pearson's correlation coefficient for explorative analysis of correlations, analysis of variance for group comparison, and chi(2)-test for investigation of distribution of 2 or more categorical variables. RESULTS: Of the 511 participants, 73% felt that CME supported them in keeping up to date with new medical knowledge, yet only half believed in its ability to minimize medical errors or increase quality of patient care. Traditional ways of learning were preferred. Significant differences in CME behavior were found between primary-care physicians and specialists, the latter using more communicative media. Also, a small group of very skeptical physicians who do not agree with the need for CME in general was identified. Average expenditure for CME was estimated between 500 and 1000 Euros per year. A majority of physicians called for other parties, eg, health insurances or government, to share the financial responsibility for CME. DISCUSSION: Mandatory CME was accepted as a necessity already before the new legislation took effect, with prevailing skepticism at the same time. Future research is needed on how to motivate physicians and especially certain subgroups of physicians to use CME methods shown to be effective in changing clinical behavior. PMID- 19998476 TI - Genetic modification of pigs as organ donors for xenotransplantation. AB - Transgenic pigs are promising donor organisms for xenotransplantation as they share many anatomical and physiological characteristics with humans. The most profound barrier to pig-to-primate xenotransplantation is the rejection of the grafted organ by a cascade of immune mechanisms commonly referred to as hyperacute rejection (HAR), acute humoral xenograft rejection (AHXR), immune cell mediated rejection, and chronic rejection. Various strategies for the genetic modification of pigs facilitate tailoring them to be donors for organ transplantation. Genetically modified pigs lacking alpha-1,3-Gal epitopes, the major xenoantigens triggering HAR of pig-to-primate xenografts, are considered to be the basis for further genetic modifications that can address other rejection mechanisms and incompatibilities between the porcine and primate blood coagulation systems. These modifications include expression of human complement regulatory proteins, CD39, endothelial protein C receptor, heme oxygenase 1, thrombomodulin, tissue factor pathway inhibitor as well as modulators of the cellular immune system such as human TNF alpha-related apoptosis inducing ligand, HLA-E/beta-2-microglobulin, and CTLA-4Ig. In addition, transgenic strategies have been developed to reduce the potential risk of infections by endogenous porcine retroviruses. The protective efficacy of all these strategies is strictly dependent on a sufficiently high expression level of the respective factors with the required spatial distribution. This review provides an overview of the transgenic approaches that have been used to generate donor pigs for xenotransplantation, as well as their biological effects in in vitro tests and in preclinical transplantation studies. A future challenge will be to combine the most important and efficient genetic modifications in multi-transgenic pigs for clinical xenotransplantation. PMID- 19998477 TI - Poly(ADP-ribose)polymerase inhibitor can attenuate the neuronal death after 1 methyl-4-phenyl-1,2,3,6-tetrahydropyridine-induced neurotoxicity in mice. AB - An excessive expression of poly(ADP-ribose)polymerase (PARP) has been demonstrated to play a key role in the pathogenesis of Parkinson's disease (PD). Here we investigated the therapeutic effect of the PARP inhibitor benzamide against 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) neurotoxicity in mice. In our HPLC and Western blot analysis, pretreatment with benzamide showed a neuroprotective effect against MPTP neurotoxicity in mice. Posttreatment with benzamide also attenuated MPTP neurotoxicity in mice. Furthermore, our immunohistochemical study showed that posttreatment with benzamide significantly prevented neuronal damage by suppressing overexpression of neuronal, microglial, and astroglial PARP after MPTP treatment. These findings have important implications for the therapeutic time window and choice of PARP inhibitors in PD patients. Our present findings provide further evidence that PARP inhibitor may offer a novel therapeutic strategy for PD. PMID- 19998478 TI - Functional silica nanoparticle-mediated neuronal membrane sealing following traumatic spinal cord injury. AB - The mechanical damage to neurons and their processes induced by spinal cord injury (SCI) causes a progressive cascade of pathophysiological events beginning with the derangement of ionic equilibrium and collapse of membrane permeability. This leads to a cumulative deterioration of neurons, axons, and the tissue architecture of the cord. We have previously shown that the application of the hydrophilic polymer polyethylene glycol (PEG) following spinal cord or brain injury can rapidly restore membrane integrity, reduce oxidative stress, restore impaired axonal conductivity, and mediate functional recovery in rats, guinea pigs, and dogs. However there are limits to both the concentration and the molecular weight of the application that do not permit the broadest recovery across an injured animal population. In this study, PEG-decorated silica nanoparticles (PSiNPs) sealed cells, as shown by the significantly reduced leakage of lactate dehydrogenase from damaged cells compared with uncoated particles or PEG alone. Further in vivo tests showed that PSiNPs also significantly reduced the formation of reactive oxygen species and the process of lipid peroxidation of the membrane. Fabrication of PSiNPs containing embedded dyes also revealed targeting of the particles to damaged, but not undamaged, spinal cord tissues. In an in vivo crush/contusion model of guinea pig SCI, every animal but one injected with PSiNPs recovered conduction through the cord lesion, whereas none of the control animals did. These findings suggest that the use of multifunctional nanoparticles may offer a novel treatment approach for spinal cord injury, traumatic brain injury, and possibly neurodegenerative disorders. PMID- 19998479 TI - It is time to study the costs and benefits of regulating continuing medical education in the United States. AB - There is a dual system of continuing medical education (CME) accreditation in the United States, with developmental milestones dating back four decades. The Accreditation Council for Continuing Medical Education (ACCME) approves national providers of CME. State medical societies (SMS) approve intrastate providers. The ACCME recognizes SMS to approve intrastate providers of CME. Both types of accredited providers, national and intrastate, designate the same credit, American Medical Association, Physician Recognition Award Category 1 Credit, which is trademarked by the AMA. The ACCME collects data to help the AMA monitor use of AMA PRA Category 1 Credit. The AMA is a nationally accredited provider of the ACCME. The AMA retains the right to act unilaterally to remove the privilege of any organization to designate Category 1 Credit. Although intrastate providers typically are charged through SMS, both types of providers, national and intrastate, pay fees for accreditation services provided by the ACCME. Select fee increases of the ACCME for 2009 through 2011 are estimated at $4.3M. Actions taken in June 2009 by the AMA House of Delegates suggest the value of accreditation and credit systems in the US should be studied with regard to costs and benefits that might further align CME with patients' interests and physicians' incentives. PMID- 19998481 TI - Electrical stimulation induces calcium-dependent release of NGF from cultured Schwann cells. AB - Production of nerve growth factor (NGF) from Schwann cells (SCs) progressively declines in the distal stump, if axonal regeneration is staggered across the suture site after peripheral nerve injuries. This may be an important factor limiting the outcome of nerve injury repair. Thus far, extensive efforts are devoted to modulating NGF production in cultured SCs, but little has been achieved. In the present in vitro study, electrical stimulation (ES) was attempted to stimulate cultured SCs to release NGF. Our data showed that ES was capable of enhancing NGF release from cultured SCs. An electrical field (1 Hz, 5 V/cm) caused a 4.1-fold increase in NGF release from cultured SCs. The ES-induced NGF release is calcium dependent. Depletion of extracellular or/and intracellular calcium partially/ completely abolished the ES-induced NGF release. Further pharmacological interventions showed that ES induces calcium influx through T type voltage-gated calcium channels and mobilizes calcium from 1, 4, 5 trisphosphate-sensitive stores and caffeine/ryanodine-sensitive stores, both of which contributed to the enhanced NGF release induced by ES. In addition, a calcium-triggered exocytosis mechanism was involved in the ES-induced NGF release from cultured SCs. These findings show the feasibility of using ES in stimulating SCs to release NGF, which holds great potential in promoting nerve regeneration by enhancing survival and outgrowth of damaged nerves, and is of great significance in nerve injury repair and neuronal tissue engineering. PMID- 19998480 TI - Interactions between TLR7 and TLR9 agonists and receptors regulate innate immune responses by astrocytes and microglia. AB - Toll-like receptors 7 (TLR7) and 9 (TLR9) are important mediators of innate immune responses. Both receptors are located in endosomal compartments, recognize nucleic acids, and signal via Myeloid differentiation factor 88 (MyD88). In the current study, we analyzed TLR7 and TLR9 induced activation of astrocytes and microglia, two cell types that contribute to innate immune responses in the CNS. TLR7 and TLR9 agonists induced similar cytokine profiles within each cell type. However, there were notable differences in the cytokine profile between astrocytes and microglia, including the production of the anti-inflammatory cytokine IL-10 and antiapoptotic cytokines G-CSF and IL-9 by microglia but not astrocytes. Costimulation studies demonstrated that the TLR7 agonist, imiquimod, could inhibit TLR9 agonist-induced innate immune responses, in both cell types, in a concentration-dependent manner. Surprisingly, this inhibition was not mediated by TLR7, as deficiency in TLR7 did not alter suppression of the TLR9 agonist-induced responses. The suppression of innate immune responses was also not due to an inhibition of TLR9 agonist uptake. This suggested that imiquimod suppression may be a direct effect, possibly by blocking CpG-ODN binding and/or signaling with TLR9, thus limiting cell activation. An antagonistic relationship was also observed between the two receptors in microglia, with TLR7 deficiency resulting in enhanced cytokine responses to CpG-ODN stimulation. Thus, both TLR7 and its agonist can have inhibitory effects on TLR9-induced cytokine responses in glial cells. PMID- 19998482 TI - Acute tryptophan depletion potentiates 3,4-methylenedioxymethamphetamine-induced cerebrovascular hyperperfusion in adult male Wistar rats. AB - The serotonergic (5-hydroxytryptamine; 5-HT) dysfunction found in depression may affect not only brain function (mood) but also cerebrovascular control. Similar, but possibly occult, disturbances may also be induced by 3,4 methylenedioxymethamphetamine-induced neurotoxicity (MDMA, or "ecstasy"). Acute tryptophan depletion (ATD) is widely used to identify vulnerability to depression, and we hypothesized that repeated MDMA administration would increase the sensitivity of rats to this acute serotonergic challenge. In this study, male Wistar rats were injected with MDMA (20 mg kg(-1), twice daily for 4 days) and challenged 3 weeks later with ATD, induced by intragastric administration of a nutritional mixture with tryptophan (TRP) removed. Cerebral metabolism (CMRG) and blood flow (CBF) were measured in parallel groups of animals following ATD by using quantitative [(14)C]2-deoxyglucose and [(14)C]iodoantipyrine autoradiographic techniques, respectively. A significant reduction in paroxetine binding to 5-HT transporter sites in MDMA-treated rats indicated 5HT terminal depletion, whereas the plasma TRP/sum large neutral amino acids ratio was reduced by 40% following ATD. Under all experimental conditions, the normal close correlation between CBF and metabolic demand was maintained. However, a global analysis of all brain regions revealed a significant decrease in the overall ratio of CBF to CMRG after ATD in control animals, whereas a higher ratio was observed after ATD in the MDMA-treated group. This increase in blood flow relative to cerebral metabolism suggests an ATD-induced loss of cerebrovascular tone in MDMA-treated animals that could have pathophysiological consequences and might conceivably contribute to the behavioral dysfunction of depression. PMID- 19998483 TI - Sulforaphane protects immature hippocampal neurons against death caused by exposure to hemin or to oxygen and glucose deprivation. AB - Oxidative stress is a mediator of cell death following cerebral ischemia/reperfusion and heme toxicity, which can be an important pathogenic factor in acute brain injury. Induced expression of phase II detoxification enzymes through activation of the antioxidant response element (ARE)/Nrf2 pathway has emerged as a promising approach for neuroprotection. Little is known, however, about the neuroprotective potential of this strategy against injury in immature brain cells. In this study, we tested the hypothesis that sulforaphane (SFP), a naturally occurring isothiocyanate that is also a known activator of the ARE/Nrf2 antioxidant pathway, can protect immature neurons from oxidative stress induced death. The hypothesis was tested with primary mouse hippocampal neurons exposed to either O(2) and glucose deprivation (OGD) or hemin. Treatment of immature neurons with SFP immediately after the OGD during reoxygenation was effective in protecting immature neurons from delayed cell death. Exposure of immature hippocampal neurons to hemin induced significant cell death, and both pre- and cotreatment with SFP were remarkably effective in blocking cytotoxicity. RT-PCR analysis indicated that several Nrf2-dependent cytoprotective genes, including NAD(P)H quinone oxidoreductase 1 (NQO1), heme oxygenase 1 (HO1), and glutamate-cysteine ligase modifier subunit (GCLM), which is involved in glutathione biosynthesis, were up-regulated following SFP treatment both in control neurons and following exposure to OGD and hemin. These results indicate that SFP activates the ARE/Nrf2 pathway of antioxidant defense and protects immature neurons from death caused by stress paradigms relevant to those associated with ischemic and traumatic injury to the immature brain. PMID- 19998484 TI - Human umbilical cord blood cell therapy blocks the morphological change and recruitment of CD11b-expressing, isolectin-binding proinflammatory cells after middle cerebral artery occlusion. AB - Secondary neurodegeneration resulting from stroke is mediated by delayed proinflammatory signaling and immune cell activation. Although it remains unknown which cell surface markers signify a proinflammatory phenotype, increased isolectin binding occurs on CD11b-expressing immune cells within injured brain tissue. Several reports have confirmed the efficacy of human umbilical cord blood (HUCB) cell therapy in reducing ischemic injury in rat after middle cerebral artery occlusion (MCAO), and these effects were attributed in part to dampened neuroinflammation. The present study examined the time course of lectin binding to cells of microglia/macrophage lineage within 96 hr after MCAO and whether delayed HUCB cell treatment alters the migration and/or morphological characteristics of these cells throughout the period of infarct expansion. Isolectin binding was up-regulated in response to injury, was maximal at 96 hr, and colocalized with cells that expressed the putative proinflammatory markers MMP-9 and nitric oxide. Isolectin-tagged fluorescence was also significantly increased at 72 hr and localized to greater numbers of amoeboid, CD11b-expressing cells relative to 51 hr. Treatment with 1 x 10(6) HUCB cells significantly reduced total lectin binding at 72 hr, as well as the total area occupied by lectin-tagged fluorescence at both 51 and 72 hr, relative to vehicle-treated controls. This effect was accompanied by a shift in the morphology of CD11b positive cells from amoeboid to ramified shape. These data indicate that HUCB cell therapy suppressed the recruitment of proinflammatory, isolectin-binding cells during the period of infarct expansion, thus offering a potential mechanism for the protective effects of HUCB cell therapy. PMID- 19998486 TI - Commentary on Sabine Hildebrandt, anatomy in the Third Reich: an outline. PMID- 19998485 TI - Enhanced neurogenesis in the olfactory bulb in adult mice after injury induced by acute treatment with trimethyltin. AB - In adults, the subventricular zone is known to contain undifferentiated neural progenitor cells that proliferate and generate the olfactory bulb (OB) interneurons throughout life. We earlier showed that trimethyltin (TMT) causes neuronal damage in the granular cell layer of the OB in adult mice. In the current study, we examined neurogenesis in the OB in adult mice after injury induced by acute treatment with TMT. On day 2 post-TMT treatment, enhanced incorporation of 5-bromo-2'-deoxyuridine (BrdU) was seen in the granular cell layer of the OB. Many of the BrdU-labeled cells were undifferentiated cells on day 2 post-treatment. On day 30 post-TMT treatment, BrdU-labeled neuronal cells were dramatically increased in number in the granular cell layer of the OB. However, TMT treatment was ineffective in affecting the migration of BrdU-labeled cells from the subventricular zone to the OB. The results of a neurosphere assay revealed that the number of neurospheres derived from the OB was significantly increased on day 2 post-TMT treatment. The neurosphere-forming neural progenitor cells derived from the OB of TMT-treated animals were capable of differentiating into neuronal cells as well as into astrocytes. Taken together, our data suggest that the OB has the ability to undergo enhanced neurogenesis following TMT induced neuronal injury in adult mice. PMID- 19998487 TI - Letter in response to Winkelmann and Schagen, Seidelman and Levi: call for a new era in research on "Medicine/Anatomy in the Third Reich". PMID- 19998488 TI - N-methyl-D-aspartate preconditioning improves short-term motor deficits outcome after mild traumatic brain injury in mice. AB - Traumatic brain injury (TBI) causes impairment of fine motor functions in humans and nonhuman mammals that often persists for months after the injury occurs. Neuroprotective strategies for prevention of the sequelae of TBI and understanding the molecular mechanisms and cellular pathways are related to the glutamatergic system. It has been suggested that cellular damage subsequent to TBI is mediated by the excitatory neurotransmitters, glutamate and aspartate, through the excessive activation of the N-methyl-D-aspartate (NMDA) receptors. Thus, preconditioning with a low dose of NMDA was used as a strategy for protection against locomotor deficits observed after TBI in mice. Male adult mice CF-1 were preconditioned with NMDA (75 mg/kg) 24 hr before the TBI induction. Under anesthesia with O(2)/N(2)O (33%: 66%) inhalation, the animals were subjected to the experimental model of trauma that occurs by the impact of a 25 g weight on the skull. Sensorimotor gating was evaluated at 1.5, 6, or 24 hr after TBI induction by using footprint and rotarod tests. Cellular damage also was assessed 24 hr after occurrence of cortical trauma. Mice preconditioned with NMDA were protected against all motor deficits revealed by footprint tests, but not those observed in rotarod tasks. Although mice showed motor deficits after TBI, no cellular damage was observed. These data corroborate the hypothesis that glutamatergic excitotoxicity, especially via NMDA receptors, contributes to severity of trauma. They also point to a putative neuroprotective mechanism induced by a sublethal dose of NMDA to improve motor behavioral deficits after TBI. PMID- 19998490 TI - Variations of the piriformis and sciatic nerve with clinical consequence: a review. AB - The deep gluteal region is often encountered when performing injections, when performing surgery such as total hip replacements, or diagnosing problems of this region or lower limbs using clinical or imaging techniques. Previously, the prevalence figures of piriformis and sciatic nerve anomalies have ranged from 1.5 to 35.8% in dissected specimens. This study systematically reviews and meta analyses the prevalence of piriformis and sciatic nerve anomalies in humans using previously published literature. A further review is conducted regarding the anatomical abnormalities present in surgical case series of procedures for patients suffering from piriformis syndrome. After pooling the results of 18 studies and 6,062 cadavers, the prevalence of the anomaly in cadavers was 16.9%; 95% confidence interval (CI) 16.0-17.9%. The prevalence of the piriformis and sciatic nerve anomaly in the surgical case series was 16.2%, 95% CI: 10.7-23.5%. The difference between the two groups was not found to be significant 0.74%; 95% CI: -5.66 to 7.13; P = 0.824. Because of the high likelihood of an anomaly being present in a patient, clinicians and surgeons should be aware of the potential complications this anomaly may have on medical or surgical interventions. Furthermore, because the prevalence of the anomaly in piriformis syndrome patients is not significantly different from what is thought to be a normal population, it indicates that this anomaly may not be as important in the pathogenesis of piriformis syndrome as previously thought. PMID- 19998489 TI - Neuroprotective effects of PDGF against oxidative stress and the signaling pathway involved. AB - The neuroprotective effects of platelet-derived growth factor (PDGF) and the major signaling pathways involved in these were examined using primary cultured mouse cortical neurons subjected to H(2)O(2)-induced oxidative stress. The specific function of the PDGF beta-receptor (PDGFR-beta) was examined by the selective deletion of the corresponding gene using the Cre-loxP system in vitro. In wild-type neurons, PDGF-BB enhanced the survival of these neurons and suppressed H(2)O(2)-induced caspase-3 activation. The prosurvival effect of PDGF AA was less than that of PDGF-BB. PDGF-BB highly activated Akt, extracellular signal-regulated kinase (ERK), c-jun amino-terminal kinase (JNK) and p38. PDGF-AA activated these molecules at lesser extent than PDGF-BB. In particular, PDGF-AA induced activation of Akt was at very low level. The neuroprotective effects of PDGF-BB were antagonized by inhibitors of phosphatidylinositol 3-kinase (PI3-K), mitogen-activated protein kinase kinase (MEK), JNK and p38. The PDGFR-beta depleted neurons showed increased vulnerability to oxidative stress, and less responsiveness to PDGF-BB-induced cytoprotection and signal activation, in which Akt activation was most strongly suppressed. After all, these results demonstrated the neuroprotective effects of PDGF and the signaling pathways involved against oxidative stress. The effects of PDGF-BB were more potent than those of PDGF-AA. This might be due to the activation and additive effects of two PDGFRs after PDGF-BB stimulation. Furthermore, the PI3-K/Akt pathway that was deduced to be preferentially activated by PDGFR-beta may explain the potent effects of PDGF-BB. PMID- 19998491 TI - Comparative structural and functional analysis of the GLT-1/EAAT-2 promoter from man and rat. AB - In the vertebrate CNS, glutamate transport predominantly occurs through the glutamate transporter subtype, GLT-1/EAAT-2, which prevails in astrocytes. GLT 1/EAAT-2 expression is impaired in many acute and chronic brain diseases, leading to increases in extracellular glutamate and subsequent excitotoxic neuronal cell death. An obvious therapeutical approach to prevent glutamate-induced brain damage would be targeting GLT-1/EAAT-2 expression. Since so far, insights into the mechanisms modulating GLT-1/EAAT-2 expression mostly originated from work with rat astrocytes, we now sought to determine whether this modulatory network would also apply to humans. To this end, we have cloned the previously unknown rat GLT-1/EAAT-2 promoter and compared it to the human promoter sequence. In reporter assays, the cloned 2.7-kb region immediately flanking the 5'-end of the rat GLT-1/EAAT-2 gene allowed for similar increases in constitutive gene expression as the human promoter sequence. Sequence analysis demonstrated the presence of highly conserved regions on the rat and human GLT-1/EAAT-2 promoters, which turned out to be likewise essential for constitutive GLT-1/EAAT-2 expression, stimulation of gene transcription by EGF, TGFalpha, and PACAP as well as inhibition of gene transcription by TNFalpha. Intriguingly, endothelin-1 which inhibits endogenous GLT-1/EAAT-2 expression, promoted activity of both rat and human reporter constructs, indicating the existence of (an) inhibitory mechanism(s) not operational in the reporter gene assay. Our findings establish close similarities in the regulation of GLT-1/EAAT-2 expression in rat and man and, hence, validate rat astrocytes as an assay system for studying the molecular mechanisms affecting glutamate homeostasis in the healthy and diseased human brain. PMID- 19998492 TI - Progressive noradrenergic deficits in the locus coeruleus of Mecp2 deficient mice. AB - Methyl-CpG binding protein 2 (MeCP2) is a transcriptional regulator. Mutations in this gene cause a wide range of neurological disorders. Mecp2 deficiency has been previously associated to catecholaminergic dysfunctions leading to autonomic defects in the brainstem and the sympathoadrenergic system of the mouse. The present study was undertaken to determine if the locus coeruleus (LC), the main noradrenergic cell group of the brain, is affected. Using real type PCR, we found a reduction of the tyrosine hydroxylase (Th) mRNA level, the rate-limiting enzyme in catecholamine synthesis, in the whole pons of P15 (-36%), P30 (-47%) and P50 ( 42%) Mecp2 null male as well as in adult heterozygous female (-44%) mice. Using immunoquantification we did not observe any difference of the Th staining level in P30 null male mice. However at P50, we demonstrated a significant decrease in both the Th staining level (-24%), and the number of Th-positive neurons (-23%). We subsequently characterized a reduction (-28%) of the dendritic density of the Th-positive fibers surrounding the LC in P50 null male mice. In heterozygous female mice immunoquantification did not revealed significant modifications, but only a tendency towards reduction. Finally, we did not found any apoptotic neurons in the pons indicating that LC neurons are not dying but are more likely loosing their catecholaminergic phenotype. In conclusion, our results showing a progressive catecholaminergic deficit in the LC of Mecp2 deficient null male mice could open new perspectives to better understand the autonomic and cognitive deficits due to the lack of Mecp2. PMID- 19998493 TI - Chronic hyperammonemia alters the circadian rhythms of corticosteroid hormone levels and of motor activity in rats. AB - Patients with liver cirrhosis may present hepatic encephalopathy with a wide range of neurological disturbances and alterations in sleep quality and in the sleep-wake circadian rhythm. Hyperammonemia is a main contributor to the neurological alterations in hepatic encephalopathy. We have assessed, in an animal model of chronic hyperammonemia without liver failure, the effects of hyperammonemia per se on the circadian rhythms of motor activity, temperature, and plasma levels of adrenal corticosteroid hormones. Chronic hyperammonemia alters the circadian rhythms of locomotor activity and of cortisol and corticosterone levels in blood. Different types of motor activity are affected differentially. Hyperammonemia significantly alters the rhythm of spontaneous ambulatory activity, reducing strongly ambulatory counts and slightly average velocity during the night (the active phase) but not during the day, resulting in altered circadian rhythms. In contrast, hyperammonemia did not affect wheel running at all, indicating that it affects spontaneous but not voluntary activity. Vertical activity was affected only very slightly, indicating that hyperammonemia does not induce anxiety. Hyperammonemia abolished completely the circadian rhythm of corticosteroid hormones in plasma, completely eliminating the peaks of cortisol and corticosterone present in control rats at the start of the dark period. The data reported show that chronic hyperammonemia, similar to that present in patients with liver cirrhosis, alters the circadian rhythms of corticosteroid hormones and of motor activity. This suggests that hyperammonemia would be a relevant contributor to the alterations in corticosteroid hormones and in circadian rhythms in patients with liver cirrhosis. PMID- 19998494 TI - Olfactory ensheathing cell membrane properties are shaped by connectivity. AB - Olfactory ensheathing cells (OECs) have been repeatedly implicated in mediating plasticity, particularly in situ in the olfactory nerve in which they support the extension of olfactory sensory neuron (OSN) axons from the olfactory epithelium to the olfactory bulb (OB). OECs are specialized glia whose processes surround OSN axon fascicles within the olfactory nerve and across the OB surface. Despite their purported importance in promoting axon extension, and following transplants, little is known about either morphology or biophysical properties of OECs in situ. In particular, cell-cell interactions that may influence OEC function are largely unexplored. Here, we studied OEC connectivity and morphology in slice preparations, preserving tissue structure and cell-cell interactions. Our analyses showed that OECs form a matrix of cellular projections surrounding axons, unique among glia, and express high levels of connexin-43. Lucifer Yellow injections revealed selective dye coupling among small subgroups of OECs. Two types of OECs were biophysically distinguished with whole-cell voltage-clamp recordings: (1) with low-input resistance (R(i)), linear current profiles, and frequently dye coupled; and (2) with high R(i), nonlinear current profiles, and infrequent dye coupling. Pharmacological blockade of gap junctions changed OEC membrane properties such that linear OECs became nonlinear. Double recordings indicated that the appearance of the nonlinear current profile was associated with the loss of electrical coupling between OECs. We conclude that the diversity of OEC current profiles can be explained by differences in gap-junction connectivity and discuss implications of this diversity for OEC influences on axon growth and excitability. PMID- 19998496 TI - Confocal laser endomicroscopy in the "in vivo" histological diagnosis of the gastrointestinal tract. AB - Recent technological advances in miniaturization have allowed for a confocal scanning microscope to be integrated into a conventional flexible endoscope, or into trans-endoscopic probes, a technique now known as confocal endomicroscopy or confocal laser endomicroscopy. This newly-developed technology has enabled endoscopists to collect real-time in vivo histological images or "virtual biopsies" of the gastrointestinal mucosa during endoscopy, and has stimulated significant interest in the application of this technique in clinical gastroenterology. This review aims to evaluate the current data on the technical aspects and the utility of this new technology in clinical gastroenterology and its potential impact in the future, particularly in the screening or surveillance of gastrointestinal neoplasia. PMID- 19998495 TI - Clinical relevance and public health significance of hepatitis B virus genomic variations. AB - Ten hepatitis B virus (HBV) genotypes (A-J) and 34 HBV subgenotypes have been identified so far. HBV genotypes and subgenotypes have distinct geographical distributions, and have been shown to differ with regard to clinical outcome, prognosis, and response to interferon treatment. Infection with subgenotype A2 is frequently associated with high viral load, resulting in acute infection via horizontal transmission. Genotypes A and B are more sensitive to interferon treatment than genotypes D and C, respectively. Genotype B is more frequent in acute hepatitis than genotype C, whereas genotype C (C2) is more frequently associated with an increased risk of hepatocellular carcinoma (HCC), mostly cirrhotic, as compared with genotype B (B2). Genotype mixture is associated with high viral load and worse outcome of HBV infection. HBV mutations in the S genes, especially amino acids substitution at position 145 (G145R), are associated with immune escape, whereas mutations in the PreS or S genes which impair HBsAg secretion could present a risk to blood safety. HBV variants harboring mutations in the viral polymerase gene that confer resistance to nucleoside analogs may be selected during antiviral therapy. Different genotypes have distinct mutation patterns in the PreS and EnhII/BCP/Precore regions. PreS deletions, C1653T, T1753V, and A1762T/G1764A are associated with an increased risk of HCC. HCC associated HBV mutants may not transmit via mother-to-child transmission, and are likely generated during HBV-induced pathogenesis. Examination of HBV mutations alone or in combination and host genetic susceptibility will be helpful in classifying the HBV-infected subjects who will develop HCC and need active anti viral treatments. PMID- 19998498 TI - Potential role of Th17 cells in the pathogenesis of inflammatory bowel disease. AB - The etiopathology of inflammatory bowel disease (IBD) remains elusive. Accumulating evidence suggests that the abnormality of innate and adaptive immunity responses plays an important role in intestinal inflammation. IBD including Crohn's disease (CD) and ulcerative colitis (UC) is a chronic inflammatory disease of the gastrointestinal tract, which is implicated in an inappropriate and overactive mucosal immune response to luminal flora. Traditionally, CD is regarded as a Th1-mediated inflammatory disorder while UC is regarded as a Th2-like disease. Recently, Th17 cells were identified as a new subset of T helper cells unrelated to Th1 or Th2 cells, and several cytokines [e.g. interleukin (IL)-21, IL-23] are involved in regulating their activation and differentiation. They not only play an important role in host defense against extracellular pathogens, but are also associated with the development of autoimmunity and inflammatory response such as IBD. The identification of Th17 cells helps us to explain some of the anomalies seen in the Th1/Th2 axis and has broadened our understanding of the immunopathological effects of Th17 cells in the development of IBD. PMID- 19998497 TI - Signal molecule-mediated hepatic cell communication during liver regeneration. AB - Liver regeneration is a complex and well-orchestrated process, during which hepatic cells are activated to produce large signal molecules in response to liver injury or mass reduction. These signal molecules, in turn, set up the connections and cross-talk among liver cells to promote hepatic recovery. In this review, we endeavor to summarize the network of signal molecules that mediates hepatic cell communication in the regulation of liver regeneration. PMID- 19998499 TI - Role of the receptor for advanced glycation end products in hepatic fibrosis. AB - AIM: To study the role of advanced glycation end products (AGE) and their specific receptor (RAGE) in the pathogenesis of liver fibrogenesis. METHODS: In vitro RAGE expression and extracellular matrix-related gene expression in both rat and human hepatic stellate cells (HSC) were measured after stimulation with the two RAGE ligands, advanced glycation end product-bovine serum albumin (AGE BSA) and N(epsilon)-(carboxymethyl) lysine (CML)-BSA, or with tumor necrosis factor-alpha (TNF-alpha). In vivo RAGE expression was examined in models of hepatic fibrosis induced by bile duct ligation or thioacetamide. The effects of AGE-BSA and CML-BSA on HSC proliferation, signal transduction and profibrogenic gene expression were studied in vitro. RESULTS: In hepatic fibrosis, RAGE expression was enhanced in activated HSC, and also in endothelial cells, inflammatory cells and activated bile duct epithelia. HSC expressed RAGE which was upregulated after stimulation with AGE-BSA, CML-BSA, and TNF-alpha. RAGE stimulation with AGE-BSA and CML-BSA did not alter HSC proliferation, apoptosis, fibrogenic signal transduction and fibrosis- or fibrolysis-related gene expression, except for marginal upregulation of procollagen alpha1(I) mRNA by AGE BSA. CONCLUSION: Despite upregulation of RAGE in activated HSC, RAGE stimulation by AGE does not alter their fibrogenic activation. Therefore, RAGE does not contribute directly to hepatic fibrogenesis. PMID- 19998500 TI - Identification of TRPM7 channels in human intestinal interstitial cells of Cajal. AB - AIM: To investigate the characteristics of slow electrical waves and the presence of transient receptor potential melastatin-type 7 (TRPM7) in the human gastrointestinal (GI) tract. METHODS: Conventional microelectrode techniques were used to record intracellular electrical responses from human GI smooth muscle tissue. Immunohistochemistry was used to identify TRPM7 channels in interstitial cells of Cajal (ICCs). RESULTS: The human GI tract generated slow electrical waves and had ICCs which functioned as pacemaker cells. Flufenamic acid, a nonselective cation channel blocker, and 2-APB (2-aminoethoxydiphenyl borate) and La(3+), TRPM7 channel blockers, inhibited the slow waves. Also, TRPM7 channels were expressed in ICCs in human tissue. CONCLUSION: These results suggest that the human GI tract generates slow waves and that TRPM7 channels expressed in the ICCs may be involved in the generation of the slow waves. PMID- 19998501 TI - Role of diffusion-weighted magnetic resonance imaging in the differential diagnosis of focal hepatic lesions. AB - AIM: To evaluate the utility of diffusion-weighted imaging (DWI) in screening and differential diagnosis of benign and malignant focal hepatic lesions. METHODS: Magnetic resonance imaging (MRI) examinations were performed using the Signa Excite Xl Twin Speed 1.5T system (GE Healthcare, Milwaukee, WI, USA). Seventy patients who had undergone MRI of the liver [29 hepatocellular carcinomas (HCC), four cholangiocarcinomas, 34 metastatic liver cancers, 10 hemangiomas, and eight cysts] between April 2004 and August 2008 were retrospectively evaluated. Visualization of lesions, relative contrast ratio (RCR), and apparent diffusion coefficient (ADC) were compared between benign and malignant lesions on DWI. Superparamagnetic iron oxide (SPIO) was administered to 59 patients, and RCR was compared pre- and post-administration. RESULTS: DWI showed higher contrast between malignant lesions (especially in multiple small metastatic cancers) and surrounding liver parenchyma than did contrast-enhanced computed tomography. ADCs (mean +/- SD x 10(-3) mm(2)/s) were significantly lower (P < 0.05) in malignant lesions (HCC: 1.31 +/- 0.28 and liver metastasis: 1.11 +/- 0.22) and were significantly higher in benign lesions (hemangioma: 1.84 +/- 0.37 and cyst: 2.61 +/- 0.45) than in the surrounding hepatic tissues. RCR between malignant lesions and surrounding hepatic tissues significantly improved after SPIO administration, but RCRs in benign lesions were not improved. CONCLUSION: DWI is a simple and sensitive method for screening focal hepatic lesions and is useful for differential diagnosis. PMID- 19998502 TI - NT4(Si)-p53(N15)-antennapedia induces cell death in a human hepatocellular carcinoma cell line. AB - AIM: To construct the recombinant lentivirus expression plasmid, pLenti6/V5-NT4 p53(N15)-antennapedia (Ant), and study its effect on HepG2 cells. METHODS: Plasmid pLenti6/V5-NT4 p53(N15)-Ant was constructed incorporating the following functional regions, including signal peptide sequence and pro-region of neurotrophin 4, N-terminal residues 12-26 of p53 and 17 amino acid drosophila carrier protein, Ant. Hepatocellular carcinoma (HepG2) cells were used for transfection. 3-[4,5-dimethyl-thiazol-2yl]-2,5 diphenyl tetrazolium bromide (MTT) assay, lactate dehydrogenase (LDH) release assay, transmission electron microscopy (TEM) and flow cytometric analysis (FCM) were employed to investigate the effects of LV-NT4(Si)-p53(N15)-Ant in vitro on HepG2 cells. In vivo experiment was also performed to investigate the inhibitory effect of LV-NT4(Si) p53(N15)-Ant on tumor growth in nude mice. RESULTS: LV-NT4(Si)-p53(N15)-Ant significantly suppressed the growth of HepG2 cells. MTT assay showed that the growth of HepG2 cells was mucj more significantly inhibited by LV-NT4(Si) p53(N15)-Ant than by LV-EGFP. The inhibition rate for HepG2 cell growth in the two groups was 46.9% and 94.5%, respectively, 48 h after infection with LV NT4(Si)-p53(N15)-Ant, and was 33.9% and 95.8%, respectively, 72 h after infection with LV-NT4(Si)-p53(N15)-Ant (P < 0.01). Light microscopy and TEM showed morphological changes in HepG2 cells infected with LV-NT4(Si)-p53(N15)-Ant, but no significant changes in HepG2 cells infected with LV-EGFP. Changes were observed in ultra-structure of HepG2 cells infected with LV-NT4(Si)-p53(N15)-Ant, with degraded membranes, resulting in necrosis. LDH release from HepG2 cells was analyzed at 24, 48, 72 and 96 h after infection with LV-NT4(Si)-p53(N15)-Ant and LV-EGFP, which showed that LDH release was significantly higher in LV-NT4(Si) p53(N15)-Ant treatment group (682 IU/L) than in control group (45 IU/L, P < 0.01). The longer the time was after infection, the bigger the difference was in LDH release. FCM analysis showed that LV-NT4(Si)-p53(N15)-Ant could induce two different kinds of cell death: necrosis and apoptosis, with apoptosis being the minor type and necrosis being the main type, suggesting that LV-NT4(Si)-p53(N15) Ant exerts its anticancer effect on HepG2 cells by inducing necrosis. The in vivo study showed that LV-NT4(Si)-p53(N15)-Ant significantly inhibited tumor growth with an inhibition rate of 66.14% in terms of tumor size and weight. CONCLUSION: LV-NT4(Si)-p53(N15)-Ant is a novel recombinant lentivirus expression plasmid and can be used in gene therapy for cancer. PMID- 19998503 TI - Separate basolateral and apical phosphatidylcholine secretion routes in intestinally differentiated tumor cells. AB - AIM: To investigate whether the secretion of phosphatidylcholine (PC) in intestinal mucus occurs by apical secretion or via basolateral excretion and to determine its subsequent passage across the tight junctions to the apical mucus. METHODS: We addressed this question using the polarized intestinally differentiated tumor cell line CaCo-2 grown on filters to confluence in Transwell culture chambers. The released PC and sphingomyelin (Sph) from apical and basolateral media were analyzed by mass spectrometry. RESULTS: The secreted PC species were identical in both compartments indicating the same intracellular origin of PC. However, PC secretion into the basolateral compartment was more effective, and the PC:Sph ratio in the basolateral compartment was significantly higher than that in the apical compartment (8.18 +/- 1.84 vs 4.31 +/- 1.22, P = 0.01). Both pathways were temperature sensitive and were unaltered in the presence of cyclosporine. CONCLUSION: The data demonstrate the PC secretion capacity of CaCo-2 cells and indicate two separated apical and basolateral release mechanisms. PMID- 19998504 TI - Value of three-dimensional reconstructions in pancreatic carcinoma using multidetector CT: initial results. AB - AIM: To evaluate the use of three-dimensional imaging of pancreatic carcinoma using multidetector computed tomography (CT) in a prospective study. METHODS: Ten patients with suspected pancreatic tumors were examined prospectively using multidetector CT (Somatom Sensation 16, Siemens, Erlangen, Germany). The images were evaluated for the presence of a pancreatic carcinoma and invasion of the peripancreatic vessels and surrounding organs. Using the isotropic CT data sets, a three-dimensional image was created with automatic vascular analysis and semi automatic segmentation of the organs and pancreatic tumor by a radiologist. The CT examinations and the three-dimensional images were presented to the surgeon directly before and during the patient's operation using the Medical Imaging Interaction Toolkit-based software "ReLiver". Immediately after surgery, the value of the two images was judged by the surgeon. The operation and the histological results served as the gold standard. RESULTS: Nine patients had a pancreatic carcinoma (all pT3), and one patient had a serous cystadenoma. One tumor infiltrated the superior mesenteric vein. The infiltration was correctly evaluated. All carcinomas were resectable. In comparison to the CT image with axial and coronal reconstructions, the three-dimensional image was judged by the surgeons as better for operation planning and consistently described as useful. CONCLUSION: A 3D-image of the pancreas represents an invaluable aid to the surgeon. However, the 3D-software must be further developed in order to be integrated into daily clinical routine. PMID- 19998505 TI - Iodized oil uptake assessment with cone-beam CT in chemoembolization of small hepatocellular carcinomas. AB - AIM: To evaluate the utility of assessing iodized oil uptake with cone-beam computed tomography (CT) in transarterial chemoembolization (TACE) for small hepatocellular carcinoma (HCC). METHODS: Cone-beam CT provided by a biplane flat panel detector angiography suite was performed on eighteen patients (sixteen men and two women; 41-76 years; mean age, 58.9 years) directly after TACE for small HCC (26 nodules under 30 mm; mean diameter, 11.9 mm; range, 5-28 mm). The pre procedural locations of the tumors were evaluated using triphasic multi-detector row helical computed tomography (MDCT). The tumor locations on MDCT and the iodized oil uptake by the tumors were analyzed on cone-beam CT and on spot image directly after the procedures. RESULTS: All lesions on preprocedural MDCT were detected using iodized oil uptake in the lesions on cone-beam CT (sensitivity 100%, 26/26). Spot image depicted iodized oil uptake in 22 of the lesions (sensitivity 85%). The degree of iodized oil uptake was overestimated (9%, 2/22) or underestimated (14%, 3/22) on spot image in five nodules compared with that of cone-beam CT. CONCLUSION: Cone-beam CT is a useful and convenient tool for assessing the iodized oil uptake of small hepatic tumors (< 3 cm) directly after TACE. PMID- 19998506 TI - Image-guided conservative management of right colonic diverticulitis. AB - AIM: To study the clinical outcomes of medical therapy in patients with right colonic diverticulitis. METHODS: The records of 189 patients with right colonic diverticulitis which was finally diagnosed by computed tomography, ultrasonography, or operative findings were retrospectively reviewed. RESULTS: Of the 189 patients hospitalized for right colonic diverticulitis, the stages of diverticulitis by a modified Hinchey classification were 26 patients (13.8%) in stage 0, 139 patients (73.5%) in stage Ia, 23 patients (12.2%) in stage Ib, and 1 patient (0.5%) in stage III. Medical therapy was undertaken in 185 of 189 patients (97.9%). One hundred and eighty three of 185 patients were successfully treated with bowel rest and antibiotics. Two patients in stage Ib required a resection or surgical drainage because of an inadequate response to conservative treatment. Recurrent diverticulitis developed in 15 of 183 patients (8.2%) who responded to medical therapy. All 15 patients who suffered a second attack had uncomplicated diverticulitis, and were successfully treated with medical therapy. CONCLUSION: Our results indicate that right colonic diverticulitis is essentially benign and image-guided conservative treatment is primarily required. PMID- 19998507 TI - Changes in intestinal mucosal immune barrier in rats with endotoxemia. AB - AIM: To investigate the dysfunction of the immunological barrier of the intestinal mucosa during endotoxemia and to elucidate the potential mechanism of this dysfunction. METHODS: Male Wistar rats were randomly distributed into two groups: control group and lipopolysaccharide (LPS) group. Endotoxemia was induced by a single caudal venous injection of LPS. Animals were sacrificed in batches 2, 6, 12 and 24 h after LPS infusion. The number of microfold (M)-cells, dendritic cells (DCs), CD4(+) T cells, CD8(+) T cells, regulatory T (Tr) cells and IgA(+) B cells in the intestinal mucosa were counted after immunohistochemical staining. Apoptotic lymphocytes were counted after TUNEL staining. The levels of interleukin (IL)-4, interferon (IFN)-gamma and forkhead box P3 (Foxp3) in mucosal homogenates were measured by ELISA. The secretory IgA (sIgA) content in the total protein of one milligram of small intestinal mucus was detected using a radioimmunological assay. RESULTS: This research demonstrated that LPS-induced endotoxemia results in small intestinal mucosa injury. The number of M-cells, DCs, CD8(+) T cells, and IgA(+) B cells were decreased while Tr cell and apoptotic lymphocyte numbers were increased significantly. The number of CD4(+) T cells increased in the early stages and then slightly decreased by 24 h. The level of IL-4 significantly increased in the early stages and then reversed by the end of the study period. The level of IFN-gamma increased slightly in the early stages and then decreased markedly by the 24 h time point. Level of Foxp3 increased whereas sIgA level decreased. CONCLUSION: Mucosal immune dysfunction forms part of the intestinal barrier injury during endotoxemia. The increased number and function of Tr cells as well as lymphocyte apoptosis result in mucosal immunodeficiency. PMID- 19998508 TI - Effect of implanting fibrin sealant with ropivacaine on pain after laparoscopic cholecystectomy. AB - AIM: To investigate the safety and efficacy of implanting fibrin sealant with sustained-release ropivacaine in the gallbladder bed for pain after laparoscopic cholecystectomy (LC). METHODS: Sixty patients (American Society of Anesthesiologists physical status was I or II and underwent LC) were randomly divided into three equal groups: group A (implantation of fibrin sealant in the gallbladder bed), group B (implantation of fibrin sealant carrying ropivacaine in the gallbladder bed), and group C (normal saline in the gallbladder bed). Postoperative pain was evaluated, and pain relief was assessed by visual analog scale (VAS) scoring. RESULTS: The findings showed that 81.7% of patients had visceral pain, 50% experienced parietal, and 26.7% reported shoulder pain after LC. Visceral pain was significantly less in group B patients than in the other groups (P < 0.05), and only one patient in this group experienced shoulder pain. The mean VAS score in group B patients was lower than that in the other groups. CONCLUSION: Visceral pain is prominent after LC and can be effectively controlled by implanting fibrin sealant combined with ropivacaine in the gallbladder bed. PMID- 19998509 TI - Multidrug resistance protein 3 R652G may reduce susceptibility to idiopathic infant cholestasis. AB - AIM: To evaluate the role of genetic factors in the pathogenesis of idiopathic infant cholestasis. METHODS: We performed a case-control study, including 78 infants with idiopathic infant cholestasis and 113 healthy infants as controls. Genomic DNA was extracted from peripheral venous blood leukocytes using phenol chloroform methodology. Polymerase chain reaction was used to amplify the multidrug resistance protein 3 (MDR3) R652G fragment, and products were sequenced using the ABI 3100 Sequencer. RESULTS: The R652G single nucleotide polymorphism (SNP) was significantly more frequent in healthy infants (allele frequency 8.0%) than in patients (allele frequency 2.60%) (P < 0.05), odds ratio, 0.29; 95% confidence interval, 0.12-0.84. The conjugated bilirubin in patients with the AG genotype was significantly lower than in those with the AA genotype (44.70 +/- 6.15 micromol/L vs 95.52 +/- 5.93 micromol/L, P < 0.05). CONCLUSION: MDR3 R652G is negatively correlated with idiopathic infant cholestasis. Children with the R652G SNP in Guangxi of China may have reduced susceptibility to infant intrahepatic cholestasis. PMID- 19998510 TI - Two synchronous somatostatinomas of the duodenum and pancreatic head in one patient. AB - Somatostatinomas are extremely rare neuroendocrine tumors of the gastrointestinal tract, first described in the pancreas in 1977 and in the duodenum in 1979. They may be functional and cause somatostatinoma or inhibitory syndrome, but more frequently are non-functioning pancreatic endocrine tumors that produce somatostatin alone. They are usually single, malignant, large lesions, frequently associated with metastases, and generally with poor prognosis. We present the unique case of a 57-year-old woman with two synchronous non-functioning somatostatinomas, one solid duodenal lesion and one cystic lesion within the head of the pancreas, that were successfully resected with a pylorus-preserving Whipple's procedure. No secondaries were found in the liver, or in any of the removed regional lymph nodes. The patient had an uneventful recovery, and remains well and symptom-free at 18 mo postoperatively. This is an extremely rare case of a patient with two synchronous somatostatinomas of the duodenum and the pancreas. The condition is discussed with reference to the literature. PMID- 19998511 TI - Is iron overload in alcohol-related cirrhosis mediated by hepcidin? AB - In this case report we describe the relationship between ferritin levels and hepcidin in a patient with alcohol-related spur cell anemia who underwent liver transplantation. We demonstrate a reciprocal relationship between serum or urinary hepcidin and serum ferritin, which indicates that inadequate hepcidin production by the diseased liver is associated with elevated serum ferritin. The ferritin level falls with increasing hepcidin production after transplantation. Neither inflammatory indices (IL6) nor erythropoietin appear to be related to hepcidin expression in this case. We suggest that inappropriately low hepcidin production by the cirrhotic liver may contribute substantially to elevated tissue iron stores in cirrhosis and speculate that hepcidin replacement in these patients may be of therapeutic benefit in the future. PMID- 19998512 TI - Neoadjuvant peptide receptor radionuclide therapy for an inoperable neuroendocrine pancreatic tumor. AB - Pancreatic endocrine tumors are rare but are among the most common neuroendocrine neoplasms of the abdomen. At diagnosis many of them are already advanced and difficult to treat. We report on an initially inoperable malignant pancreatic endocrine tumor in a 33-year-old woman, who received neoadjuvant peptide receptor radionuclide therapy (PRRT) as first-line treatment. This resulted in a significant downstaging of the tumor and allowed its subsequent complete surgical removal. Follow-up for eighteen months revealed a complete remission. This is the first report on neoadjuvant PRRT in a neuroendocrine neoplasm with subsequent successful complete resection. PMID- 19998513 TI - Cronkhite-Canada syndrome associated with myelodysplastic syndrome. AB - We report a case of Cronkhite-Canada syndrome (CCS) associated with myelodysplastic syndrome (MDS). A 54-year-old woman, diagnosed as MDS the prior year after evaluation of anemia, visited our hospital with the chief complaint of epigastric discomfort. She also had dysgeusia, alopecia, atrophic nail change, and pigmentation of the palm, all of which began several months ago. Blood tests revealed severe hypoalbuminemia. Colonoscopy (CS) showed numerous, dense, red polyps throughout the colon and rectum. Biopsy specimens showed stromal edema, infiltration of lymphocytes, and cystic dilatation of the crypt. Her clinical manifestations and histology were consistent with CCS. We prescribed corticosteroids, which dramatically improved her physical findings, laboratory data, and endoscopic findings. This is the first report of CCS in a patient with MDS. PMID- 19998514 TI - Spontaneous liver rupture in hypereosinophilic syndrome: a rare but fatal complication. AB - We report a rare case of spontaneous liver rupture in a patient with hypereosinophilic syndrome (HES), of which the diagnosis was delayed, resulting in a fatal outcome. The diagnostic criteria and treatment of HES with hepatic involvement were reviewed. The possible cause of spontaneous liver rupture in HES and its management were also discussed. To our knowledge, this is the first case report of spontaneous liver rupture in HES. We emphasized the need of a high index of suspicion in diagnosing HES, so that early treatment could be initiated. PMID- 19998515 TI - Surgery for rare aneurysm associated with colorectal cancer. AB - The occurrence of concomitant aortic aneurysm and colorectal cancer is a rare medical entity, and controversy surrounds its optimal treatment. We report a case of rectal cancer and concomitant aneurysm from the ascending aorta to the common iliac artery. As with DeBakey type I aortic dissecting aneurysm, our patient was treated by rectal cancer resection, with preservation of the anus (Dixon operation) under controlled hypotension. Blood pressure was maintained at 80 90/50-60 mmHg and the pulse at 70-90 beats/min. The pathological examination of the surgical specimen showed a poorly differentiated T3N0 tumor. The patient had an uneventful recovery without aneurysm rupture, and was discharged from hospital on postoperative day 15 after 3 d adjuvant chemotherapy with oxaliplatin combined with calcium folinate and fluorouracil. The patient was given six courses of adjuvant chemotherapy in 6 mo, without recurrence or metastasis, and the aneurysm was still stable after 2 years follow-up. PMID- 19998516 TI - Lethal neuroendocrine carcinoma in ulcerative colitis. AB - A 48-year old male with longstanding and extensive pancolitis developed a high grade and rapidly lethal malignant lesion in the ascending colon characterized by a neuroendocrine carcinoma. Prior biopsies obtained from multiple sites in the colon during endoscopic surveillance were reported to show only inflammatory changes without dysplasia. Although operator-dependent, repeated endoscopic studies may have limitations during surveillance programs because the biological behavior of some colonic neoplastic lesions may have a rapid and very aggressive clinical course. PMID- 19998517 TI - Editorial comment. PMID- 19998518 TI - Editorial comment. PMID- 19998519 TI - Editorial comment. PMID- 19998520 TI - Proceedings of the 4th International Conference on Spatial Cognition: Spatial Cognition and Action. Rome, Italy. September 14-18, 2009. PMID- 19998521 TI - A review of human carcinogens--Part F: chemical agents and related occupations. PMID- 19998522 TI - Natural products and healthy heart. PMID- 19998523 TI - Antioxidant properties of an endogenous thiol: Alpha-lipoic acid, useful in the prevention of cardiovascular diseases. AB - In the past few years, a growing interest has been given to the possible antioxidant functions of a natural acid, synthesized in human tissues: alpha lipoic acid (ALA). Both the oxidized (disulfide) and reduced (dithiol: dihydrolipoic acid, DHLA) forms of ALA show antioxidant properties. ALA administered in the diet accumulates in tissues, and a substantial part is converted to DHLA via a lipoamide dehydrogenase. Commercial ALA is usually a racemic mixture of the R and S forms. Chemical studies have indicated that ALA scavenges hydroxyl radicals, hypochlorous acid, and singlet oxygen. ALA exerts antioxidant effects in biological systems not only through direct ROS quenching but also via transition metal chelation. ALA has been shown to possess a number of beneficial effects both in the prevention and treatment of diabetes in experimental conditions. ALA presents beneficial effects in the management of symptomatic diabetic neuropathy and has been used in this context in Germany for more than 30 years. In cardiovascular disease, dietary supplementation with ALA has been successfully employed in a variety of in vivo models: ischemia reperfusion, heart failure, and hypertension. More mechanistic and human in vivo studies are needed to determine whether optimizing the dietary intake of ALA can help to decrease cardiovascular diseases. A more complete understanding of cellular biochemical events that influence oxidative damage is required to guide future therapeutic advances. PMID- 19998524 TI - Dispute over pneumococcal vaccine initiative. PMID- 19998525 TI - The long shadow of malaria interventions in tropical Africa. PMID- 19998526 TI - Exploratory safety pharmacology: a new safety paradigm to de-risk drug candidates prior to selection for regulatory science investigations. AB - The primary objective of Safety Pharmacology is to ensure the safety of medicines on physiological functions in order to protect humans against adverse drug reactions. Safety Pharmacology became a major non-clinical discipline in 2000 when the International Conference on Harmonization approved the S7A guideline. This regulatory document requires pharmaceutical companies to undertake Safety Pharmacology assessment under Good Laboratory Practice (GLP) in order to guarantee the absence of unmanageable risks on vital organ function for compounds to be tested on humans. These regulatory studies often reveal liabilities impacting on the smooth transition of drug candidates from the discovery phase into the clinical arena. However, if these safety issues were uncovered prior to regulatory science assessment, the chemistry of poorly safe molecules could be modified during the lead optimisation phase for preventing later occurring attrition accidents. This article proposes the establishment of a spin-off specialty of Regulatory Safety Pharmacology, for which the name 'Exploratory Safety Pharmacology' is proposed. The objective of this discipline would be to conduct early safety investigations on potential drug candidates by applying, outside the constraints of GLP, in silico, in vitro, ex vivo and in vivo platforms translating clinical liabilities into simple, fast and cost-effective screening assays. This approach should result in early hazard detection with rapid turnaround of the data, enabling medicinal chemists to mitigate the safety liabilities of new compounds in an iterative manner. Hence, the ultimate aim of Exploratory Safety Pharmacology activities is to transform Regulatory Safety Pharmacology investigations into risk-known exercises. PMID- 19998527 TI - Orlistat: current status in clinical therapeutics. AB - BACKGROUND: Obesity has rapidly become a life-threatening epidemic worldwide. There are a plethora of obesity-related co-morbidities and complications that increase morbidity, mortality and cost of care associated with obesity. Orlistat is approved for the treatment of obesity and has been evaluated both in obesity and in the several obesity-related co-morbidities. OBJECTIVE: The purpose of this article is to provide detail of the pharmacotherapeutic role of orlistat in obesity, describe orlistat and its pharmacological properties, critique the evidence for orlistat's use in obesity and obesity related co-morbidities, and define the role of orlistat in clinical practice. METHODS: A thorough, all inclusive literature search was conducted to isolate clinical trials, case reports and meta-analyses evaluating the safety and efficacy of orlistat in various patient populations. RESULTS: Orlistat's unique mechanism of action, beneficial effects on multiple co-morbidity surrogates and relatively mild adverse effect and drug interaction profile position it favorably as the first option for pharmacotherapy in comprehensive obesity management of adults and children. PMID- 19998528 TI - Octreotide LAR: safety and tolerability issues. AB - BACKGROUND: Somatostatin analogues are the cornerstone in therapy of acromegaly and functioning neuroendocrine tumors. Long-acting retard formulations have improved patient survival and contributed considerably to quality of life. The first such compound was octreotide LAR ('long-acting release'), characterized by high affinity to somatostatin receptor subtypes 2 and 5, which has to be injected intramuscularly every 4 weeks. OBJECTIVE: The aim was to screen all octreotide LAR-related literature and assess the compound's profile for safety and tolerability. METHODS: An extensive literature search has been performed using the MEDLINE database to retrieve data from clinical studies evaluating the efficacy and tolerability of octreotide LAR. RESULTS/CONCLUSION: Octreotide LAR is well tolerated; however, diarrhea and gallstone formation were identified as the main adverse events. Impairment of glucose homeostasis was a regular phenomenon, but its occurrence was unpredictable. General side effects such as headache, abdominal discomfort or fatigue were also reported. According to incidental case reports, administration during pregnancy appears to be safe for both mother and child; however, definitive evidence is missing. In addition, octreotide LAR has been evaluated for further indications including treatment of solid tumor entities, due to its antiproliferative effect. Currently, several compounds (lanreotide, SOM230) with a broader receptor spectrum are under evaluation and may improve treatment efficacy and lower incidence of side effects. PMID- 19998529 TI - A review of acitretin for the treatment of psoriasis. AB - BACKGROUND: Acitretin is an oral retinoid that is approved for the treatment of psoriasis. It is unique compared to other systemic therapies for psoriasis such as methotrexate and cyclosporine in that it is not immunosuppressive. It is, therefore, safe for use in psoriasis patients with a history of chronic infection such as HIV, hepatitis B, hepatitis C or malignancy who have a contraindication to systemic immunosuppressive therapy and require systemic therapy because topical therapy is inadequate and they are unable to commit to phototherapy. Acitretin is one of the treatments of choice for pustular psoriasis. Even though acitretin is less effective as a monotherapy for chronic plaque psoriasis, combination therapy with other agents, especially UVB or psoralen plus UVA phototherapy, can enhance efficacy. OBJECTIVE: To provide an updated review of the safety and efficacy of acitretin in the treatment for psoriasis. METHODS: Literature review of journal articles from 2008 to 2009 since the last review of acitretin evaluated medical literature from 2005 to 2008. RESULTS/CONCLUSION: Acitretin is an effective systemic therapy for psoriasis and is generally well tolerated at low doses for long-term use. If monotherapy with acitretin is inadequate, it can be used in combination with other treatments, particularly UVB phototherapy, to increase efficacy. PMID- 19998530 TI - Self-consciousness, friendship quality, and adolescent internalizing problems. AB - The correlates between public and private self-consciousness and internalizing difficulties were examined during early adolescence. Friendship quality was assessed as a possible moderator of the relation between self-consciousness and maladjustment. One hundred and thirty-seven young adolescents (N = 87girls; M age = 13.98 years) reported on their self-consciousness, internalizing problems, and the quality of their best friendship. Results indicated stronger associations between private self-consciousness and internalizing correlates than between public self-consciousness and internalizing problems, suggesting that private self-consciousness may be a stronger risk factor during adolescence. Contrary to expectations, evidence revealed that positive friendship quality may exacerbate some difficulties associated with self-consciousness. Results pertaining to friendship quality add to the growing literature on the ways in which friendships can contribute to adjustment difficulties. PMID- 19998531 TI - The effects of modelling, local stimulus enhancement, and affordance demonstration on the production of object-directed actions in 6-month-old infants. AB - The present study investigated whether local stimulus enhancement and the demonstration of objects' affordances--both of which are inherent in modelling of object-directed target actions--are themselves sufficient to evoke the target behaviour on imitation test trials. Six-month-old infants were presented with a puppet wearing a removable mitten and observed either a demonstration of mitten removal (modelling group), the experimenter pointing at the mitten (stimulus enhancement group), the mitten falling off apparently by itself (affordance demonstration group), or no specific action directed at the mitten (control group). For all infants, the puppet was next presented without any accompanying demonstrations and infants' mitten removal behaviours in the response period were recorded. The results showed that local stimulus enhancement and affordance demonstration were as effective as full modelling in evoking the target action. This finding shows that the performances of 6-month-old infants on imitation tests can be multiply determined and evoked by variables other than modelling of target behaviour. PMID- 19998532 TI - Childhood rituals and executive functions. AB - Repetitive and ritualistic behaviours (RRBs) are a feature of both typical and atypical development. While the cognitive correlates of these behaviours have been investigated in some neurodevelopmental conditions these links remain largely unexplored in typical development. The current study examined the relationship between RRBs and executive functions (EF) in a sample of typically developing children aged between 37 and 107 months. Results showed that cognitive flexibility, and not response inhibition or generativity, was most strongly associated with the frequency of RRBs in this sample. In younger children (<67.5 months) cognitive flexibility was significantly associated with 'Repetitive Behaviours' but in older children (>67.5 months) cognitive flexibility was associated with both 'Just Right' and 'Repetitive Behaviour', suggesting that the association between EF and RRBs may become stronger with age in typically developing children. PMID- 19998533 TI - From fancy to reason: scaling deaf and hearing children's understanding of theory of mind and pretence. AB - We examined deaf and hearing children's progression of steps in theory of mind (ToM) development including their understanding of social pretending. Ninety three children (33 deaf; 60 hearing) aged 3-13 years were tested on a set of six closely matched ToM tasks. Results showed that deaf children were delayed substantially behind hearing children in understanding pretending, false belief (FB) and other ToM concepts, in line with their delayed uptake of social pretend (SP) play. By using a scaling methodology, we confirmed previous evidence of a consistent five-step developmental progression for both groups. Moreover, by including social pretence understanding, both deaf and hearing children's ToM sequences were shown to extend reliably to six sequential developmental steps. Finally and focally, even though both groups' sequences were six steps long, the placement of pretence relative to other ToM milestones varied with hearing status. Deaf children understood social pretending at an earlier step in the ToM sequence than hearing children, albeit at a later chronological age. Theoretically, the findings are relevant to questions about how universal developmental progressions come together along with culturally distinctive inputs and biological factors (such as hearing loss) to set the pace for ToM development. PMID- 19998534 TI - Attentional processes in young children with congenital visual impairment. AB - The study investigated attentional processes of 32 preschool children with congenital visual impairment (VI). Children with profound visual impairment (PVI) and severe visual impairment (SVI) were compared to a group of typically developing sighted children in their ability to respond to adult directed attention in terms of establishing, maintaining, and shifting attention on toys. The measures of children's sensory-motor understanding (SMU) and language ability were obtained using the Reynell-Zinkin scales of mental development. The videorecordings of these play-based assessments were coded for three categories of behavioural responses (Establish, Maintain, and Shift). The three groups were matched on verbal comprehension (VC), but differed significantly in their SMU and their chronological age. The groups of children with PVI and SVI were found to be comparable in their ability to establish and maintain attention on objects. Despite a relatively good performance overall both groups scored significantly lower on those skills than children who were sighted. However, with regards to attention shifting, children with PVI showed significantly lower performance than both the children with SVI and the sighted children who were similar on this component. Ability to maintain and shift attention was significantly related to the cognitive ability of children with PVI; however the poorer attentional responses were not confined only to the children with low IQ. The results are discussed in relation to the role of vision, cognitive ability and executive function in attentional processes in children with congenital VI. PMID- 19998535 TI - Early tracking of informant accuracy and inaccuracy. AB - Three- and four-year-old children (N = 131) were tested for their sensitivity to the accuracy and inaccuracy of informants. Children were presented with one of three conditions. In the Accurate-Inaccurate condition, one informant named objects accurately whereas the other named them inaccurately. In the Accurate Neutral condition, one informant named objects accurately whereas the other merely drew attention to them. Finally in the Inaccurate-Neutral condition, one informant named objects inaccurately whereas the other merely drew attention to them. In subsequent test trials, 4-year-olds preferred to seek and accept information in a selective fashion across all three conditions, suggesting that they monitor informants for both accuracy and inaccuracy. By contrast, 3-year olds were selective in the Accurate-Inaccurate and Inaccurate-Neutral conditions but not in the Accurate-Neutral condition, suggesting that they monitor informants only for inaccuracy and take accuracy for granted. PMID- 19998537 TI - Young children understand multiple pretend identities in their object play. AB - This set of studies examined the ability of 3-year-olds to conceptualize multiple pretend identities with objects. Rather than relying on verbal response measures, as has been done in the past, children's creative and inferential pretend actions were used as indicators of their understanding. The common structure to all four studies was that children were confronted with one pretend scenario, moved to a second pretend scenario and then back again to the first. Children proficiently tailored their pretence to an object whose pretend identity changed between scenarios despite being less able to name each identity. Thus, using an inferential action methodology, these studies provide early and particularly convincing evidence that children can track the multiple pretend identities of objects. PMID- 19998536 TI - When hearts, hands, and feet trump brains: centralist versus peripheralist responses in children and adults. AB - A series of studies examined the presence of centralist versus peripheralist responding about the physical location of psychological processes. Centralists respond that processes such as cognition and emotion are a function of the brain. Peripheralists respond that such processes are located in other parts of the body, such as the heart. Although peripheralist responses declined across grade levels, even older children and adults often gave peripheralist answers, depending on the context of the questions. Peripheralist responses occurred when participants were asked about the effect of switching irrelevant body parts between two people and when they were asked to choose a different body part among four choices. Results also showed adults' responses varied with different contextual cues. The findings support a coexistence model of development which argues for the simultaneous presence of developmentally advanced reasoning or scientifically based knowledge along with presumably less advanced, intuitive based reasoning, or folk beliefs. PMID- 19998538 TI - An investigation of the impact of young children's self-knowledge of trustworthiness on school adjustment: a test of the realistic self-knowledge and positive illusion models. AB - The study aimed to examine the relationship between self-knowledge of trustworthiness and young children's school adjustment. One hundred and seventy three (84 male and 89 female) children from school years 1 and 2 in the United Kingdom (mean age 6 years 2 months) were tested twice over 1-year. Children's trustworthiness was assessed using: (a) self-report at Time 1 and Time 2; (b) peers reports at Time 1 and Time 2; and (c) teacher-reports at Time 2. School adjustment was assessed by child-rated school-liking and the Short-Form Teacher Rating Scale of School Adjustment (Short-Form TRSSA). Longitudinal quadratic relationships were found between school adjustment and children's self-knowledge, using peer-reported trustworthiness as a reference: more accurate self-knowledge of trustworthiness predicted increases in school adjustment. Comparable concurrent quadratic relationships were found between teacher-rated school adjustment and children's self-knowledge, using teacher-reported trustworthiness as a reference, at Time 2. The findings support the conclusion that young children's psychosocial adjustment is best accounted for by the realistic self knowledge model (Colvin & Block, 1994). PMID- 19998539 TI - Generativity and imagination in autism spectrum disorder: evidence from individual differences in children's impossible entity drawings. AB - This study examined the cognitive underpinnings of spontaneous imagination in autism spectrum disorder (ASD) by way of individual differences. Children with ASD (N = 27) and matched typically developing (TD) children were administered Karmiloff-Smith's (1990) imaginative drawing task, along with measures that tapped specific executive functions (generativity, visuospatial planning, and central coherence processing style) and false belief theory of mind (ToM) understanding. The ASD group drawings displayed deficits in imaginative content and a piecemeal pictorial style. ASD participants also showed group deficits in generativity, planning and ToM, and exhibited weak coherence. Individual differences in generativity were related to imaginative drawing content in the ASD group, and the association was mediated through planning ability. Variations in weak coherence were separately related to a piecemeal drawing style in the ASD group. Variations in generativity were also linked with imaginative drawing content in the TD group; the connection unfolded when it received pooled variance from receptive language ability, and thereupon mediated through false belief reasoning to cue imaginative content. Results are discussed in terms of how generativity plays a broad and important role for imagination in ASD and typical development, albeit in different ways. PMID- 19998540 TI - Young children's understanding of the context-relativity of normative rules in conventional games. AB - We investigated young children's awareness of the context-relative rule structure of simple games. Two contexts were established in the form of spatial locations. Familiar objects were used in their conventional way at location 1, but acquired specific functions in a rule game at location 2. A third party then performed the conventional act at either of the two locations, constituting a mistake at location 2 (experimental condition), but appropriate at location 1 (control condition). Three-year-olds (but not 2-year-olds) systematically distinguished the two conditions, spontaneously intervening with normative protest against the third party act in the experimental, but not in the control condition. Young children thus understand context-specific rules even when the context marking is non-linguistic. These results are discussed in the broader context of the development of social cognition and cultural learning. PMID- 19998541 TI - Relationships between magical thinking, obsessive-compulsiveness and other forms of anxiety in a sample of non-clinical children. AB - Despite the obvious phenomenological similarities between magical thinking and obsessive-compulsiveness, the relationship between them has been the subject of few empirical investigations in samples of children. The present study aimed to examine the relationship between a general epistemic stance towards magical causation and tendencies towards obsessive-compulsiveness in a non-clinical sample of schoolchildren. One-hundred and two children, aged between 5 and 10 years (48 boys and 54 girls), completed questionnaire measures designed to assess magical thinking, obsessive-compulsiveness, and other forms of anxiety. School teachers completed a measure of strengths and difficulties for each child. General belief in magical causation was correlated with all types of anxiety, not just obsessive-compulsiveness, with significant correlations shown for boys in the sample, but not girls. General belief in magical causation contributed little to the prediction of obsessive-compulsiveness beyond general anxiety. In this study, a general epistemic stance towards magical causation did not differentiate obsessive-compulsiveness from other anxiety dimensions. The findings are considered in the context of developmental theories of magical and scientific causal reasoning. PMID- 19998542 TI - An examination of emotion regulation, temperament, and parenting style as potential predictors of adolescent depression risk status: a correlational study. AB - Given that depression is a debilitating disorder, it is critical that we advance our understanding about the aetiology of this disorder. This study investigated both traditional (temperament and parenting) and novel (emotion regulation strategy) risk factors associated with adolescent depression. Forty-four adolescents (12-16 years; 64% females) with high scores on a self-report depressive symptomatology questionnaire were compared to a similar group of 44 adolescents with low scores, matched for age, gender, and ethnicity. Significant group differences were present on all assessed risk factors. The presence of high depressive symptomatology was found to be associated with (1) low levels of temperamentally based positive mood, flexibility, and approach behaviours, (2) a parenting style characterized by low nurturance and high overprotection, and (3) emotion regulation characterized by higher levels of expressive suppression and lower levels of cognitive reappraisal. It was concluded that, in addition to specific temperament characteristics and parenting style, use of particular emotion regulation strategies is associated with varying levels of depressive symptomatology. These findings reinforce the importance of incorporating emotion regulation into explanatory models of depression symptomatology. Further research that tests the direction of effects for these cross-sectional findings is warranted. PMID- 19998543 TI - Childhood anxiety and attention to emotion faces in a modified stroop task. AB - This study used an emotional face stroop task to investigate the effects of self report trait anxiety, social concern (SC), and chronological age (CA) on reaction time to match coloured outlines of angry, happy, and neutral faces (and control faces with scrambled features) with coloured buttons in a community sample of 74 children aged 6-12 years. The results showed an interference of colour matching for angry (relative to neutral) faces in children with elevated SC. The same effect was not found for happy or control faces. In addition, the results suggest that selective attention to angry faces in children with social concern (SC) was not significantly moderated by age. PMID- 19998544 TI - Speech priming: an examination of rate and syntactic persistence in preschoolers. AB - Interactional coordination is important for conversational competence. For example, the syntactic form and rate of perceived speech can influence future productions in adults. Previous work has shown that children are similarly primed by syntax. This experiment demonstrates that syntactic priming and rate priming exist simultaneously in children. Participants (4- and 5-years-old) alternated between listening to priming sentences that described visual scenes and producing their own descriptions of similar scenes. The priming sentences varied in rate (fast and slow) and syntactic structure (active and passive). Children's sentences reflected the timing and syntactic structure of the primes, and there were developmental differences in their rate persistence. PMID- 19998545 TI - Is understanding regret dependent on developments in counterfactual thinking? AB - Children's understanding of counterfactual emotions such as regret and relief develops relatively late compared to their ability to imagine counterfactual worlds. We tested whether a late development in counterfactual thinking: understanding counterfactuals as possibilities, underpinned children's understanding of regret. Thirty 5- and 6-year-olds completed tasks assessing counterfactual thinking and understanding regret. Performance on the counterfactual task was better than that on the regret task. We suggest that thinking about counterfactuals as possibilities is a necessary but not sufficient cognitive development in children's understanding of regret. We discuss how other developments in counterfactual thinking may underpin children's emotional understanding. PMID- 19998546 TI - [Towards a better outcome for children]. PMID- 19998547 TI - [Childhood cancers, progress and perspectives]. PMID- 19998548 TI - Fertile grounds. PMID- 19998549 TI - Violence against women. Guest editor's introduction. PMID- 19998550 TI - [Primary sclerosing cholangitis]. PMID- 19998551 TI - Molecular advances in early breast cancer. Breast cancers are as diverse as the women who have them. Now experts are using gene-based information to tailor treatment. PMID- 19998552 TI - Becoming a vegetarian. Avoiding meat is only one part of the picture. A healthy vegetarian diet should be chock-full of foods with known benefits. PMID- 19998553 TI - Study shows that a new type of osteoporosis drug reduces fracture risk. PMID- 19998554 TI - Psychodynamic therapy is effective in treating chronic worry. PMID- 19998555 TI - By the way, doctor. I have stress urinary incontinence and don't want to have surgery. What can you tell me about Renessa? PMID- 19998556 TI - Recognizing and managing ADHD in adults. Symptoms tend to evolve and attenuate with age, but may still require treatment. PMID- 19998557 TI - Supporting survivors of suicide loss. Family, friends, co-workers, and mental health clinicians are all affected. PMID- 19998558 TI - Treating somatoform disorders. Most research supports cognitive behavioral therapy, but other options exist. PMID- 19998559 TI - One question may help screen for unhealthy alcohol use. PMID- 19998560 TI - Commentary. The Mediterranean diet, physical activity, and cognitive health. PMID- 19998562 TI - [On the (pre)history of ethnological encyclopedia production activities and the present status of Slovenian ethnological encyclopedia scholarship]. PMID- 19998563 TI - [Serbian economic, cultural, educational, and humanitarian organizations]. PMID- 19998564 TI - [The structure of peasant families in the regions of Prigorje and Zagorje, 1918 1941]. PMID- 19998565 TI - Why health reform will bend the cost curve. AB - The health reform bills passed by the U.S. House of Representatives and under consideration in the Senate introduce a range of payment and delivery system changes designed to achieve a significant slowing of health care cost growth. Most assessments of health reform legislation have focused only on the federal budgetary impact. This study projects the effect of national reform on total national health expenditures and the insurance premiums that American families would likely pay. We estimate that the combination of provisions in the House and Senate bills would save $683 billion or more in national health spending over the 10-year period 2010-2019 and lower premiums by nearly $2,000 per family. Moreover, the annual growth rate in national health expenditures could be slowed from 6.4 percent to 6.0 percent PMID- 19998566 TI - Empirically based treatments for family caregiver distress: what works and where do we go from here? AB - Family caregivers of older adults with significant cognitive and/or physical impairment are at increased risk for both psychiatric and physical morbidity. This article examines the research literature dedicated to the development of effective interventions to reduce distress and enhance well-being for these caregivers. Using a recent empirically based treatment (EBT) review of the literature as a backdrop, 3 overarching types of interventions were described as effective: psychoeducational skill building, psychotherapy (cognitive-behavioral in focus), and multicomponent (using a combination of at least 2 approaches such as education, family meetings, and skill building). Suggestions are made to facilitate future caregiver intervention research and translation of EBTs into the community including the need to expand rigorous research with caregivers of older adults facing problems other than dementia, including older patients with psychiatric problems; develop and test interventions designed for transitions into and out of the caregiving roles, as well as interventions designed to improve physical health outcomes and promote health behaviors; create linkages between interventions at multiple levels of delivery; extend caregiver intervention work with underrepresented ethnic and racial populations; and investigate the cost-effectiveness of caregiver interventions. PMID- 19998567 TI - Climate change is almost totally nature-driven. PMID- 19998568 TI - In a time of torture. PMID- 19998569 TI - [The sea is the place of the origin of life on our planet]. PMID- 19998570 TI - The future: telemedicine. PMID- 19998571 TI - Problems associated with direct-to-consumer advertising (DTCA) of restricted, implantable medical devices: should the current regulatory approach be changed? AB - Advertising and promotion of Food and Drug Administration (FDA)-approved medical products has been one of the most controversial and bitterly litigated areas in food and drug law in the U.S. for more than a decade. Hundreds of newspaper articles and dozens of law review articles have been written on the subject of the risks and benefits of direct to consumer advertising (DTCA) of medical products, but until very recently virtually all of this literature and commentary has focused exclusively on prescription and over-the-counter drugs. Even when FDA has sponsored public hearings to address the issue of DTCA of all medical products, as it did in 2005, review of the content of the speakers' presentations reveals that almost all of the subject matter, nearly all of the data, and the majority of comments concerned DTCA of drugs. Not a single law review article has ever been devoted exclusively to the subject of advertising and promotion of medical devices to consumers--until now. PMID- 19998572 TI - Dietary supplement labeling and advertising claims: are clinical studies on the full product required? AB - Whether labeling and advertising claims for multi-ingredient dietary supplements may be based on the testing of individual, key ingredients--rather than the actual product--has caused a good deal of confusion. This confusion stems from the dearth of case law and the open-endedness of Federal Trade Commission (FTC) and Food and Drug Administration (FDA) guidance on this issue. Nevertheless, the relevant regulatory guidance, case law and self-regulatory case law--when assessed together--indicate that the law allows and even protects "key ingredient claims" (i.e., claims based on efficacy testing of key ingredients in the absence of full product testing). This article provides an overview of the relevant substantiation requirements for dietary supplement claims and then reviews FTC's and FDA's guidance on key ingredient claims; relevant case law; use of key ingredient claims in the advertising of other consumer products; and the National Advertising Division of the Better Business Bureau, Inc.'s (NAD's) approach to evaluating key ingredient claims for dietary supplements. This article concludes that key ingredient claims--provided they are presented in a truthful and non deceptive manner--are permissible, and should be upheld in litigation and cases subject to industry self-regulation. This article further concludes that the NAD's approach to key ingredient claims provides practical guidance for crafting and substantiating dietary supplement key ingredient claims. PMID- 19998573 TI - Pandemic influenza: is there a corporate duty to prepare? AB - This article considers whether in the wake of an influenza pandemic companies may be exposed to claims of legal liability for failing to provide employees with access to antiviral medications, as the Department of Health and Human Services (HHS) now encourages businesses to do. It begins by describing influenza and influenza pandemics. It then discusses the benefits and limitations of antiviral therapies and the recent creation of antiviral option programs. It concludes by considering whether claims may be brought on the theory that corporate leadership is under a duty to prepare for a pandemic by considering whether to provide access to antiviral protection for employees. PMID- 19998574 TI - Government regulation of pricing and reimbursement of prescription medicines: results of a recent multi-country review. AB - Access to prescription medicines is an essential component of publicly funded or publicly assisted healthcare programs. Thus, important policy questions are presented when government agencies that administer those programs establish procedures for making decisions about the appropriate use of medicines, including determinations as to which medicines will be reimbursed, at what prices, and for which patients. The authors reviewed systems for making determinations on pricing and reimbursement of prescription medicines under public healthcare systems in 10 countries, including four member states of the European Union (EU) (France, Germany, Italy and the United Kingdom), plus Australia, Brazil, China, India, Japan and Korea. In addition to national regulatory requirements, the authors considered international agreements that may impose obligations with respect to procedures for pricing and reimbursement of prescription medicines, including the EU's Transparency Directive, agreements administered by the World Trade Organization (WTO), and bilateral free trade agreements negotiated by the United States in recent years. Drawing on this experience, the authors sought to identify common themes, pitfalls and best practices in national systems for regulating pricing and reimbursement of prescription medicines. The authors have focused primarily on procedures, rather than the underlying political assumptions or substantive economic objectives. They have assumed that all countries have--or should have--a common interest in assuring that decision-making procedures afford interested persons who have relevant information a meaningful opportunity to participate PMID- 19998575 TI - Memory improving and antidepressant effects of topiramate in rats. AB - Topiramate (TOP, CAS 97240-79-4) particularly potentiates gamma-aminobutyric acid (GABA) neuroinhibition, and GABA and glutamate receptors, which have also been implicated in memory formation. Patients' giving up treatment due to adverse effects (disorders of attention, memory, and verbal fluency) is the main problem with a therapy based on this drug. The antidepressant effect of TOP administered to rats in the dose of 15 mg/kg is observed only after 14 and 21 days of treatment. The delay in the antidepressant effect of TOP may be due to the modulation of GABA A receptors as well as due to the influence of the drug on receptors for type AMPA/KAIN excitatory amino acids. The research has also shown that long-term treatment with TOP in a dose of 15 mg/kg improved spatial memory, as tested in rats with Morris test. Probably, this results from the influence of the drug on GABA A receptors and the function of glutaminergic receptors (especially in processes like coding, retrieving, and potentiation of information), adaptive processes in the brain, notwithstanding, being an important factor. As TOP is a normothymic drug, its use in the treatment of epilepsy may also positively influence cognitive processes in the so-called interseizure intervals with memory disorders; the same effects can be expected in the treatments of bipolar disorder. PMID- 19998576 TI - Gender differences in the pharmacokinetics of rivastigmine in rats. AB - The effect of gender on the pharmacokinetics of rivastigmine (CAS 123441-03-2) was studied in male and female Wistar rats following intravenous bolus administration. The area under the plasma concentration-time curve (AUC), apparent volume of distribution (Vd), systemic clearance (CL), and terminal plasma halflife (t1/2) of rivastigmine were compared between male and female rats. Compared to male rats, female rats exhibited higher plasma rivastigmine levels showing significantly (p < 0.05) larger AUC (226.77 vs. 149.68 ng h/ml), Vd (6.70 vs. 4.13 L), t1/2 (0.84 vs. 0.34 h) and a lower CL (5.51 vs. 8.35 L/h). The male rats had a 2.5 fold greater elimination rate constant than female rats (2.02 vs. 0.82 h(-1)). Gender had a significant effect on the pharmacokinetics of rivastigmine. Gender differences were reported due to gonadal hormones, and the observed difference in pharmacokinetics of rivastigmine might be attributed to testosterone in male rats. PMID- 19998577 TI - Effects of DA-6034, a flavonoid derivative, on mucin-like glycoprotein and ocular surface integrity in a rabbit model. AB - This study was designed to assess whether DA-6034 (7-carboxymethyloxy-3',4',5 trimethoxy flavone monohydrate), a new synthetic derivative of eupatilin, increases secretion of mucin-like glycoprotein and some mucins species in conjunctiva and cornea, and contributes to the preservation of ocular surface integrity. Human conjunctival and corneal epithelial cells were incubated with DA 6034 (1-250 microM). To investigate mucin secreting activity more directly, isolated rat conjunctival goblet cells were also used. Corneal protection was investigated using a desiccation-induced rabbit model of dry eye syndrome. It was found that DA-6034 increased mucin-like glycoprotein levels of both conjunctival and corneal epithelial cells at concentrations above 100 microM. Using human conjunctival epithelial cells, it was demonstrated that treatment with DA-6034 (200 microM) significantly increased production of some mucins species including MUC1, MUC2, MUC4, MUC5AC, MUC5B, and MUC16. DA-6034 also significantly increased MUC5AC production from conjunctival goblet cells isolated from rats. In the rabbit desiccation model, an ophthalmic suspension containing 3% DA-6034 significantly reduced corneal damage induced by desiccation. These results suggest that DA-6034 is a good candidate for treatment of dry eye through maintaining ocular surface integrity, which might be related to mucin secretion. PMID- 19998578 TI - Therapeutic properties and tolerance of procaine and phenazone containing ear drops in infants and very young children. AB - Since ear pain is often intolerable in very young children with acute otitis media (AOM), a safe pain reduction is indispensable both from the medical as well as from the ethical point of view. METHODS: To confirm the safety and therapeutic properties of ear drops (Otalgan) consisting of the short acting local anesthetic procaine hydrochloride (CAS 51-05-8) and the anti-inflammatory component phenazone (CAS 60-80-0), a multi-center, non-interventional post marketing surveillance (PMS) study was performed in 428 children (f/m: 45.6%/ 54.4%) aged between 0 and 6 years (mean 3.44 ys). Otalgia as the overall diagnostic criterion present in all children was allotted to otitis media in 398 (93%) and to otitis externa in 28 patients (6.5%), while in two patients, both diagnoses were documented. RESULTS: No adverse drug reactions (ADR) to the target medication have been detected during the entire study. The evaluation of tolerance by the physicians (mean score 1.38) and by the children/ parents (mean score 1.48) was in compliance with the absence of ADR. The drug tolerance was rated by the physicians in 99.8% of the cases as very good (62.2%; n = 266) or good (37.6%; n = 161). In one child (0.2%), the tolerance was evaluated by the physician as moderate. The judgement of parents revealed for all children either a very good (51.9%; n = 222) or good (48.1%; n = 206) evaluation of tolerance. The target medication was tolerated well, independent of the duration of treatment (2 to 10 days with a median of 5 days), of the daily dose and of the accumulated dose per treatment cycle, i.e. longer treatments and higher doses were not associated with a worsening of tolerance evaluations. Even in the highest dose class of over 100 drops per treatment cycle, no changes of tolerance could be revealed. As far as can be derived from non-controlled observational data, the results also confirm the known efficacy profile of the target medication in children. Under the treatment, the initial mean pain score of 2.33 was reduced by nearly 93% down to 0.17, accompanied by a corresponding normalization of the otoscopic findings of the ear canal and the tympanic membrane, and by the Improvement of the general condition. In 95.3% of the children, the physicians rated the efficacy of the target medication as very good (n = 156; 36.4%) or good (n = 252; 58.9%) while in 14 patients (3.3%) the efficacy was scored as moderate, in 4 patients (0.9%) as minimal and in two patients as inadequate (0.5%). The results also demonstrate that with the use of the target medication for local symptomatic treatment of pain in the outer ear canal and in AOM in children, the use of antibiotics in most of the cases can be avoided. CONCLUSIONS: The findings are in full accordance with the results of former clinical studies, with positive evaluations of ototoxicity and with accumulated pharmacovigilance data showing that since introduction of the drug in 1911, no ADR have ever been reported in children. The study confirms that the target medication provides a safe and rapid pain reduction and improvement of inflammation in very young children suffering from painful acute inflammatory ear conditions. PMID- 19998579 TI - Study on the pharmacokinetics of synthetic genistein after multiple oral intake in post-menopausal women. AB - Genistein (CAS 446-72-0), an isoflavone and phytoestrogen predominantly found in soy, is considered a promising natural bioactive to prevent post-menopausal bone loss. geniVida (previously Bonistein), a novel product containing of min. 98.5% synthetic genistein aglycone, was investigated in 12 healthy post-menopausal women to assess the safety and tolerability as well as to obtain pharmacokinetic data after 7 days of repeated intakes. 24 h pharmacokinetic profiles were determined after the first oral dose and after 7 days repeated intakes of 30 mg of the test formulation. Plasma genistein (aglycone) and its conjugates were determined by a standardised LC/MS analytical method using D4-genistein as the internal standard. The plasma-concentration time profiles for conjugated genistein showed a fast, monophasic one-peak course until T(max) (5.9 h (first dose), 5.3 h (steady state (SS)); C(max) (456.8 ng/ml (first dose), 498.5 ng/ ml (SS)). Elimination half-lives (t1/2) were calculated to be 10.8 h (first dose) and 8.2 h (SS), respectively. Determination of AUC(0-inf.)) (first dose) was good with a low percentage of extrapolation (3949.1 h ng/ ml). AUC(0-24h) at SS was 5923.3 h ng/ml. Steady state was reached after 4 to 5 days and no relevant accumulation occurred (R = 1.02). The test formulation was safe and very well tolerated. PMID- 19998581 TI - Investigation of selective cytotoxicity and determination of ligand induced apoptosis of a new acenaphtho [1,2-b] quinoxaline derivative. AB - Several new acenaphtho[1,2-b]quinoxaline derivatives were prepared by the reaction of o-phenylenediamines with acenaphthenequinones. The response of different carcinoid cell lines to these compounds were evaluated by MTT (3-(4,5 dimethylthiazol-2-yl)2,5-diphenyl tetrazolium bromide) assay and trypan blue exclusion tests. The cytotoxicity of 3,4-dinitroacenaphtho[1,2-b]quinoxaline (IIId) on the tested cell lines was confirmed by both tests. Furthermore, the MTT test showed a significant difference (p < 0.05) between the cytotoxicity of this compound on malignant cell lines of Caco-2, HT-29, T47D and non malignant mouse fibroblast cell line of NIH-3T3. An apoptosis inducing effect of compound IIId on K562 cells was detected by flow cytometry using Annexin-V-fluorescein isothiocyanate (AnV-FITC) and propidium iodide (PI) staining. The apoptosis induction (PI-/AnV+) in treated K562 cells was significantly (p < 0.01) more at 0.5 microg/ml concentration of compound IIId in comparison to all other concentrations of this compound and also doxorubicin (CAS 25316-40-9) (250 nM). PMID- 19998580 TI - Two-way crossover bioequivalence study of alendronate sodium tablets in healthy, non-smoking male volunteers under fasted conditions. AB - This study was conducted in order to assess the bioequivalence of two different formulations containing 70 mg alendronate sodium (CAS 121268-17-5) under fasted conditions. One hundred twenty-two healthy male volunteers were enrolled in an open label, randomized, crossover design with a wash-out period of 20 days in one study center. Urine samples were collected up to 36 h post-dose, and the concentrations of alendronic acid were determined using a high performance liquid chromatographic method with pre-derivatization and fluorescence detection (HPLC/FL) method. The mean Ae(0-t) were 604.24 +/- 348.73 microg and 627.36 +/- 327.99 microg, while the mean R(max) were 193.87 +/- 114.68 microg/h and 202.00 +/- 107.83 microg/h for the test and reference formulations, respectively. The T(max) of the test and reference tablets were 1.26 +/- 0.58 h and 1.26 +/- 0.51 h, respectively. No significant differences of pharmacokinetic parameters between the two studied formulations were found. The 90% confidence intervals for the primary target parameters, intra-individual ratios for Ae(0-t) and R(max) of alendronic acid, were between 0.86-1.00 and 0.85-1.01, respectively, and thus within the acceptance range for bioequivalence criteria. In the light of the present study it can be concluded that the test formulation is bioequivalent to the reference formulation. PMID- 19998582 TI - Bioequivalence study of two minocycline capsule formulations in healthy volunteers. AB - The bioequivalence of two capsule formulations containing 100 mg minocycline was assessed in 12 healthy adult male and female volunteers in a crossover, randomized, single-blind study. The participating volunteers were required to fast overnight and in the next morning and were given orally one capsule of the test drug (Acnez) or one capsule of the reference drug. Blood samples were drawn immediately before taking the drug (control), and at 0.33, 0.67, 1, 2, 3, 4, 6, 8, 12, 24, 36, 48, and 60 h after drug administration. One week after the first drug administration (washout period), the procedure was repeated using the alternate drug. Plasma concentrations of the drug were determined by high performance liquid chromatography method with ultraviolet detection (HPLC-UV). The pharmacokinetic parameters assessed in this study were area under the plasma concentration-time curve from time zero to 60 h (AUC(t)), area under the plasma concentration-time curve from time zero to infinity (AUC(inf)), the peak plasma concentration of the drug (C(max)), time needed to achieve the peak plasma concentration (t(max)), and the elimination half life (t1/2). The mean AUC(t), AUC(inf), C(max), and t were 18038.55 ng x h x mL(-1), 19648.21 ng x h x mL(-1), 1076.01 ng x mL(-1), and 17.33 h, respectively, for the test drug and 17979.43 ng x h x mL(-1), 19639.78 ng x h x mL(-1), 1095.97 ng x mL(-1), and 16.44 h, respectively, for the reference drug. The median (range) of t(max) of the test drug and reference drug were 2.0 (1.0-4.0) h and 2.0 (0.67-4.0) h, respectively. The geometric mean ratios of the test drug/the reference drug for AUC(t), AUC(inf), and C(max) were 98.27% 98.30%, and 97.31%, respectively. The 90% confidence intervals (CIs) were 89.26-108.19% for AUC(t), 89.95-107.41% for AUC(inf), and 89.55-105.73% for C(max). Using Wilcoxon matched-pairs test on the original data, there was no statistically significant difference found between the test and the reference drug products for t(max), values. It can be concluded that the two minocycline capsules (test drug and reference drug) are bioequivalent in terms of the rate and extent of absorption. PMID- 19998584 TI - Safeguarding the vulnerable in custody. AB - 2009 saw two deeply disturbing coronial decisions about the deaths of persons in custody in different parts of Australia. Coroner Jamieson in Victoria made telling criticisms of the quality of medical care provided to a prisoner at Port Phillip Prison who died because he was unable to summon help when the intercom system in his cell failed while he was dying of an asthma attack. State Coroner Hope in Western Australia castigated government and private facilities that played a role in causing the death by heatstroke of an indigenous man who was transported in appalling conditions to Kalgoorlie. The facts of each death raise fundamental questions about the quality of care provided to those in custody in contemporary Australia and about cultures of inadequate respect for the human rights of those in confinement. The development of a rights consciousness throughout the custodial community by charters of rights and freedoms may play a role in engendering greater respect for the imprisoned. The strengthening of coronial legislation to mandate accessibility to coronial decisions and responsiveness to them on the part of entities the subject of recommendations for improved health and safety may also play a constructive role in adding to the checks and balances against abuses of the rights of prisoners to life, humane treatment and receipt of adequate health services. PMID- 19998583 TI - Bioequivalence evaluation of a fixed dose combination lamivudine + stavudine tablet with concurrent administration of lamivudine tablet and stavudine capsule in healthy volunteers. AB - The study was designed to compare the rate and extent of absorption of a fixed dose combination tablet of lamivudine (CAS 134678-17-4) and stavudine (CAS 3056 17-5) with the concurrent administration of lamivudine tablet and stavudine capsule in 24 healthy volunteers under fasting conditions. The volunteers were randomly assigned to the test or reference treatment, with the two treatment periods separated by a washout period of at least 7 days. Plasma samples were analyzed for both analytes lamivudine and stavudine by a validated analytical method. Since the 90% confidence intervals for the "test/reference" mean ratio of the In-transformed pharmacokinetic variables C(max) AUC(0-t) and AUC(0-infinity) were clearly within the conventional bioequivalence range of 80% to 125%, the two treatments were considered bioequivalent. The safety profiles of both the test and reference formulations were comparable. PMID- 19998585 TI - Defendants' liability for pure mental harm to third parties in Australia: still a work in progress. AB - In Australia, both common and statutory law allows compensation for negligently occasioned recognised psychiatric injury, but distinguishes between pure mental harm and consequential mental harm. This column briefly discusses the concept of pure "mental harm" and the major Australian cases relating to defendants' liability to third parties for causing them pure mental harm (Jaensch v Coffey (1984) 155 CLR 549; Tame v New South Wales; Annetts v Australian Stations Pty Ltd (2002) 211 CLR 317; Sullivan v Moody (2001) 207 CLR 562; and Gifford v Strang Patrick Stevedoring Pty Ltd (2003) 214 CLR 269). The analysis focuses on judicial approaches to determining liability in these cases, especially causation. Lack of guiding principles and precise tests for attribution of liability are illustrated by Kemp v Lyell McEwin Health Service (2006) 96 SASR 192. This case is analysed first in the context of common law, and then in the light of the reform legislation contained in the Civil Liability Act 1936 (SA) and similar provisions in other jurisdictions. PMID- 19998586 TI - Disputed family claims to bury or cremate the dead. AB - A significant number of court decisions in Australia have wrestled with the issue of who among equally ranked next-of-kin should have priority in determining the timing and place of a loved one's burial or cremation. The first port of call for such decision-making can be the coroner who must determine to whom to release a body, where a death has been reportable, but disputation occurs also in non coronial contexts and has repeatedly fallen for resolution by Supreme Court judges. The decisions have identified a variety of practical considerations which have been taken into account. However, there remains considerable uncertainty about the significance of factors such as religious, spiritual and cultural values, as well as the nature and extent of the care-giving role, as influential considerations in respect of courts' decisions. While greater predictability of courts' decision-making might be therapeutic, it may be that the variability of factual situations precludes the construction of a hierarchy of relevant considerations. PMID- 19998587 TI - When two are born as one: the ethics of separating conjoined twins. AB - Conjoined twins pose a serious challenge to medical ethics because of the fact that most legally informed ethicists recognise the need to respect the sanctity of life principle for all children. This principle is not negated by impairments to a child nor by the fact that two children's fates may be so closely intertwined that what one does to one inevitably affects the other. Various mitigating factors enter clinical decisions such as the best interests of the child but these can appear rather strained in some of the more debated conjoined twins cases. We therefore need to evaluate the ethics of treatment for conjoined twins quite carefully so that significant ethical values and legal principles are not compromised or distorted beyond credibility. The idea of a potentiality principle that is close to but distinct from the substantial benefit principle as it is used in adult clinical decisions is mooted as one valuable tool in this area. PMID- 19998588 TI - National regulation and accreditation of Australian health practitioners. AB - In 2010, nurses and midwives, together with a number of other identified health professionals, will come under the Australian National Registration and Accreditation Scheme for Health Professionals. The scheme is focused on the improvement of safety and quality of health care services through the independent accreditation of educational programs and the establishment and maintenance of a national public register. In this column the framework and legislative provisions supporting the scheme are discussed. PMID- 19998589 TI - Constitutional limits on federal legislation practically compelling medical employment: Wong v Commonwealth; Selim v Professional Services Review Committee. AB - A recent decision by the High Court of Australia (Wong v Commonwealth; Selim v Professional Services Review Committee (2009) 236 CLR 573) (the PSR case) has not only clarified the scope of the Australian constitutional prohibition on "any form of civil conscription" in relation to federal legislation concerning medical or dental services (s 51xxiiiA), but has highlighted its importance as a great constitutional guarantee ensuring the mixed State-federal and public-private nature of medical service delivery in Australia. Previous decisions of the High Court have clarified that the prohibition does not prevent federal laws regulating the manner in which medical services are provided. The PSR case determined that the anti-overservicing provisions directed at bulk-billing general practitioners under Pt VAA of the Health Insurance Act 1973 (Cth) did not offend the prohibition. Importantly, the High Court also indicated that the s 51(xxiiiA) civil conscription guarantee should be construed widely and that it would invalidate federal laws requiring providers of medical and dental services (either expressly or by practical compulsion) to work for the federal government or any specified State, agency or private industrial employer. This decision is likely to restrict the capacity of any future federal government to restructure the Australian health care system, eg by implementing recommendations from the National Health and Hospitals Reform Commission for either federal government or private corporate control of presently State-run public hospitals. PMID- 19998590 TI - Science and judicial proceedings--seventy-six years on. AB - The intersection of law and science, particularly in relation to causality and the legal concept of causation, were of considerable interest to Sir Owen Dixon. In this article, revisiting Dixon's 1933 lecture "Science and Judicial Proceedings", the Chief Justice refers to Dixon's deep interest in science and the issues to which it can give rise in legal proceedings. The 1933 lecture followed shortly after the judgment of the High Court in Australian Knitting Mills Ltd v Grant (1933) 50 CLR 387 which involved consideration of expert testimony and causal connections between product characteristics and personal injury to the consumer. PMID- 19998591 TI - The influence of the pharmaceutical industry in medicine. AB - Pharmaceutical companies are known to be among the most profitable companies in the world. Proceedings of legal cases and published research provide insights into the nature of the influence of drug companies on research and publication practices relating to the drugs they manufacture, on marketing disguised as "education" and on doctors who prescribe their drugs. The influence of drug companies extends further to sponsorship of opinion leaders who promote their drugs and groups that produce clinical guidelines. More rigorous regulation of the relationship between the pharmaceutical industry and medicine is required. PMID- 19998592 TI - Determining manner of death: statistical modelling of coronial decisions. AB - Lack of standardised procedures and the varied expertise of decision-makers can make manner of death determination in ambiguous cases equivocal in its own right. The aim of the present study was to identify factors influencing coronial manner of death determinations in cases of equivocal death. Twenty-nine equivocal cases were randomly selected from the files of an experienced British county coroner. The coroner had determined these cases to be accidental deaths (n = 10), open findings (n = 8) or suicides (n = 11). Case files were individually reviewed and the information contained within each was identified for subsequent analysis. The structure of the data was examined using multidimensional scalogram analysis (MSA). Overall, MSA conducted on the data revealed cases fell within three geometric regions (accidental death, open finding and suicide), thus indicating a moderate systematic basis for manner of death determinations by this coroner. However, within the three MSA regions seven cases were identified where the coroner's manner of death determination differed from MSA manner of death region suggested by the case data. These discrepant cases are discussed in detail, with specific reference to the influence of subjective case variables on manner of death determinations. PMID- 19998593 TI - Defensive medicine in general practice: recent trends and the impact of the Civil Liability Act 2002 (NSW). AB - This article presents the results of a survey conducted among New South Wales medical practitioners to assess the extent to which the enactment of the Civil Liability Act 2002 (NSW) has reduced the practice of defensive medicine. The new legislation was intended in part to reduce the practice of defensive medicine, both "assurance-type" measures, such as performing additional tests to assure patients they have received all possible care, and "avoidance-type" measures, such as avoiding the treatment of patients who may be at a higher risk for adverse outcomes and therefore at higher risk for filing lawsuits. However, the results of the survey reveal that many medical practitioners in New South Wales remain unaware of the legal reforms and the consequent reduction in their legal liability and continue to practise defensive medicine. This article argues therefore that while the ultimate aim of reducing litigation has been achieved in New South Wales through the introduction of the Civil Liability Act, the underlying and arguably more important aim of providing medical practitioners with a more secure environment in which to practise their profession effectively has not been achieved. The apparent failure to disseminate the legal changes to the medical profession illustrates the limitations of law reform to effectively engender social change without the active use of educative and other implementation initiatives. PMID- 19998594 TI - Termination of a minor's pregnancy: critical issues for consent and the criminal law. AB - The recent Supreme Court decision of Queensland v B [2008] 2 Qd R 562 has significant implications for the law that governs consent and abortions. The judgment purports to extend the ratio of Secretary, Department of Health and Community Services (NT) v JWB and SMB (1992) 175 CLR 218 (Marion's Case) and impose a requirement of court approval for terminations of pregnancy for minors who are not Gillick-competent. This article argues against the imposition of this requirement on the ground that such an approach is an unjustifiable extension of the reasoning in Marion's Case. The decision, which is the first judicial consideration in Queensland of the position of medical terminations, also reveals systemic problems with the criminal law in that State. In concluding that the traditional legal excuse for abortions will not apply to those which are performed medically, Queensland v B provides further support for calls to reform this area of law. PMID- 19998595 TI - Worth the wait? A critique of the Abortion Act 2008 (Vic). AB - This article offers a critique of the likely impact of the Abortion Act 2008 (Vic) in light of the fact that the Act was intended to reflect rather than alter current clinical practice surrounding abortion. The author traces the development of abortion law in Victoria and compares the two models for regulating abortion: the "common law model" and the "legislative model". The author argues in favour of legislative intervention. The author also discusses current uncertainties that exist due to the unclear effect of the Charter of Human Rights and Responsibilities Act 2006 (Vic) on abortion legislation, focusing on the intersection between women's rights to an abortion and doctors' rights to freedom of conscience. PMID- 19998596 TI - Removing the "relative" uncertainty within the Australian donor insemination network. AB - In Australia there is no federal legislation limiting the use of donor sperm. However, it is important to place limits on the multiple use of sperm donors to reduce the risk of inadvertent half-sibling mating between the offspring of anonymous donors and to control for the consequences of contact between identity release donors and their donor-inseminated offspring. A nationally mandated donor registry should be established to enable, first, the calculation of updated variable values for use in the development and implementation of a predictive model to estimate the probability of half-siblings mating and provide policy makers with empirical evidence to inform the setting of anonymous donor limits; and secondly, the linking of identity-release donors to their donor-inseminated offspring and an investigation into the psychosocial consequences of that linking so as to be able to implement suitable donor limits as well as management strategies and support systems for these new "extended families" within the donor insemination network. PMID- 19998597 TI - Why the "widespread agreement" is wrong: contesting the non-harm arguments for the prohibition of full commercial surrogacy. AB - Entering a commercial surrogacy agreement is an offence in almost all Australian jurisdictions. A 2009 Consultation Paper produced by the Standing Committee of Attorneys-General suggested that there was "widespread agreement" that commercial surrogacy should remain prohibited. The arguments most commonly raised against legalising commercial surrogacy are not harm-based; that is, they do not purport to show that any party involved is tangibly, objectively and non-consensually worse off as a result of the transaction. This would be very difficult to show. Rather, the arguments against commercial surrogacy tend to focus on non-harm considerations, including principally concerns about the commodification of life and exploitation. This article argues that there are no sound non-harm reasons for banning one form of commercial surrogacy namely full commercial surrogacy. PMID- 19998598 TI - Early dental implant failure after immediate placement. PMID- 19998599 TI - Disruptive physicians. An old problem comes under new scrunity in an era of patient safety. AB - How to handle the disruptive physician or other clinician is an old and complex problem that is coming under new scrutiny in an era of patient safety. PMID- 19998600 TI - How to listen to your lawyer. AB - As regulations become more complex and tensions increase between conservative legal guidance and the move to more transparency, the issue of when and how to listen to your lawyer deserves the board's focused attention. PMID- 19998601 TI - Contrarian leadership for challenging times. PMID- 19998602 TI - Physician alignment. Long-running trend intensifies because of recession, reform fears. AB - Most hospital leaders feel strongly that their future viability depends on the organization's relationship with physicians. Hospitals are using different methods to obtain physician alignment. Also: Challenges in building the medical home. PMID- 19998603 TI - Electronic health records and the board's connection. AB - As hospitals spend millions on information technology, boards have an important oversight role to play from system selection to implementation. PMID- 19998604 TI - For the common good. AB - Hospital leaders are called not just to lead and advocate for hospitals or even health care, but for the total society of which we are members. PMID- 19998606 TI - [Climate change affects the health]. PMID- 19998605 TI - Tools to alleviate safety concerns. AB - Resources and support provided by the Agency for Healthcare Research and Quality can help trustees reduce a number of unknown factors when tackling patient safety issues. PMID- 19998607 TI - [Hope about necessary reform of health care in USA]. PMID- 19998608 TI - [Clinical research must attract the young. Universities and health care can together show possibilities of research]. PMID- 19998609 TI - [Recruitment of medical students to research requires early engagement. A research assistant program shows that early initiation yields better results]. PMID- 19998610 TI - [Self-sampling at home can prevent cervix cancer]. PMID- 19998611 TI - [Video-assisted technique is best when performing laparoscopic gastrostomy in children. Meta-analysis of the frequency of fistulas following two different methods]. PMID- 19998612 TI - [Noradrenaline infusion was the solution in severe orthostatism. Ambulatory intravenous administration turned dependence into independence]. PMID- 19998614 TI - [The National Board of Health and Welfare about the registry debate: "The registries open for unique possibilities"]. PMID- 19998613 TI - [Cochlear implant is good--but not always sufficient]. PMID- 19998615 TI - [Time for a new outpatient psychiatry]. PMID- 19998616 TI - [The Swedish Medical Association about increased medical education: The Swedish National Agency for Higher Education will assess the suitability of Orebro]. PMID- 19998617 TI - [Tycho Brahe's moustache a posthumous witness to murder?]. PMID- 19998618 TI - ODA patient's page. Gum disease. PMID- 19998619 TI - Tracking charitable care. PMID- 19998620 TI - Diffuse pigmentation of the palate. PMID- 19998621 TI - ANF campaign tell them it is 'time to deliver' in aged care. PMID- 19998622 TI - Indigenous prisoner health: where is the prevention focus? PMID- 19998623 TI - Health care and the right to cultural liberty. PMID- 19998624 TI - Nursing intervention for chronic disease management. PMID- 19998625 TI - A script for change. Will nurse practitioners be allowed to fly? PMID- 19998626 TI - Being prepared for rising temperatures. PMID- 19998627 TI - Doing time as a court liaison nurse. PMID- 19998628 TI - A hospital wide nursing documentation project. PMID- 19998629 TI - HARP--addressing chronic disease management and complex care needs in western Melbourne. PMID- 19998630 TI - NPs advancing renal care. PMID- 19998631 TI - Helping nurses achieve better practice. PMID- 19998632 TI - Collaborating to improve health outcomes for older people. PMID- 19998633 TI - Nurse led respiratory team assist GPs. PMID- 19998634 TI - Ged. Cutting Medicare rebates for cataract surgery. PMID- 19998635 TI - Major depressive disorder in the African American population: meeting the challenges of stigma, misdiagnosis, and treatment disparities. AB - This article examines major depressive disorder (MDD) in the African American population. As prevalence rates and severity of depression in African Americans are investigated, the findings indicate many blacks are underdiagnosed. Further, African Americans seem to have more severe episodes of depression compared to Caucasians. Explanations for this difference are that African Americans with MDD often present with somatic symptoms, leading physicians to miss a MDD diagnosis. Depression is often stigmatized in the African American population, seen as a "personal weakness." Educating the community about depression and educating physicians to make cultural competent diagnoses are necessary. Treatment disparities emerge as African Americans are more likely uninsured, and many are nonresponsive to traditional pharmacological interventions for depression. African American and other ethnic groups differ in the way they metabolize selective serotonin reuptake inhibitors, leading physicians to have less of an understanding of how to treat the African American patients. The lack of minorities in research trials limits the number of effective medication to treat this population of patients. PMID- 19998636 TI - Depression, suicidal ideation, and attempts in black patients with sickle cell disease. AB - There is a strong relationship between suicidal ideation, suicide attempts, and depression. Rates of successful suicides are relatively high among the chronically ill compared to other populations but are reduced with treatment. Depression and suicide rates also often differ among blacks as compared to other populations. Using survey methods, we evaluated self-reported rates of depression, suicidal ideation, and suicide attempts in 30 male and 37 female black patients with sickle cell disease (SCD). SCD is a condition characterized by chronic, unpredictable pains and psychosocial distress. Thirty-six percent of the sample self-reported depression in the past 30 days, while 22 percent of the sample exhibited scores on the Beck Depression Inventory indicative of mild or greater depression (mean BDI, 8.31 +/- 7.79). Twenty-nine percent of patients indicated an episode of suicidal ideation and 8%, a suicidal attempt in their lifetime. Thirty-three percent reported treatment by a mental health professional. We conclude that there is a continuing need for mental health services in the management of depressed affect and risk for suicide among patients with SCD. Standards of clinical care must remain flexible to accommodate the mental health needs of this population of patients. PMID- 19998638 TI - Community-based strategies for recruiting older, African Americans into a behavioral intervention study. AB - PURPOSE: To describe community-based strategies that were effective in recruiting older African Americans into a behavioral intervention study designed to increase eye examination behavior. METHODS: Sites were identified that targeted older African-Americans, including senior centers, senior housing, and church groups. We conducted presentations at these sites, networked with community organizations, placed ads on the radio and in newspapers, and attended health fairs. Potential participants also called us in response to flyers and through word of mouth. RESULTS: We conducted 147 activities at 118 sites. A total of 688 potential participants were screened, with 330 (48%) enrolling, 33% ineligible, and 19% not interested. Highest enrollment rates were for word of mouth (69%), flyers (67%), and senior centers (66%). Barriers to participation included hesitancy of seniors to leave their apartments to attend presentations and competing health issues taking precedence over eye concerns. CONCLUSIONS: A multifaceted recruitment approach incorporating both direct and indirect activities at a variety of sites should be used to recruit older African Americans into a behavioral intervention study. Establishing relationships in the community, both prior to initiating recruitment activities and as an ongoing process, was important to the study's success. PMID- 19998639 TI - Patient and provider determinants associated with the prescription of adjuvant hormonal therapies following a diagnosis of breast cancer in Medicaid-enrolled patients. AB - PURPOSE: This study examined patient and provider characteristics associated with being prescribed an aromatase inhibitor (AI) vs tamoxifen-only therapy among a cohort of postmenopausal North Carolina Medicaid enrollees diagnosed with hormone receptor-positive breast cancer. METHODS: A logistic regression model was built to determine the odds of an individual ever receiving an AI during the study period using data from the Linked North Carolina Central Cancer Registry-Medicaid Claims database. RESULTS: A total of 452 patients were included, of which 307 (67.9%) and 145 (32.1%) received tamoxifen only and AI (alone or in combination) therapy, respectively. Results of the final logistic model revealed that odds of receiving an AI generally increased over the study period; however, patients who lived in urban areas had 1.86 (95% CI, 1.20-2.89) times the odds of ever receiving some form of AI therapy compared to patients who lived in rural areas. Additionally, patients with distant stage or unstaged breast cancer (opposed to local stage) had 2.15 (95% CI, 1.39-3.32) times the odds of ever receiving an AI. CONCLUSIONS: Results suggest that the use of AI therapy is becoming more widespread over time; however, differences in the type of antiestrogenic treatment prescribed based on urban/rural status may represent disparities in access to advanced care. Furthermore, it may be the case that women with local stage breast cancer are not being treated aggressively enough with novel antiestrogenic drug therapies. PMID- 19998637 TI - A pilot investigation of the rates and predictors of mental health services utilization among depressed African American mothers and their children. AB - OBJECTIVE: Depressed mothers, especially those who are African American, are likely to underutilize mental health services. Children of depressed mothers are an at-risk population with mental health needs that are often unmet. This prospective pilot study examined 3-month frequency rates and predictors of mental health utilization for a sample of African American depressed mothers and their children. METHODS: Mothers and 1 of their children completed assessment interviews. Three months later, mothers completed a telephone interview of maternal and child mental health utilization. RESULTS: Overall, 65.3% of depressed mothers and 36.7% of their children had utilized mental health services. Logistic regression analyses indicated that initial mental health treatment significantly predicted maternal mental health utilization. Maternal reports of child behavioral problems significantly predicted child mental health utilization. CONCLUSIONS: The findings were consistent with other research and showed evidence of mental health use among African Americans who are in need of such services. PMID- 19998640 TI - A retrospective characterization of African- and European American asthmatic children in a pediatric critical care unit. AB - OBJECTIVE: To determine if African American and European American children with asthma admitted to an intensive care unit (ICU) had different characteristics, we conducted a retrospective chart review of asthma admissions to the region's only pediatric ICU. PATIENTS AND METHODS: A chart review was performed on 125 patients with asthma admitted to the pediatric critical care unit at Vanderbilt Children's Hospital. Descriptive statistics, clinical characteristics, and disparities in care were compared using either Fisher's exact tests or Wilcoxon ranksum tests. RESULTS: Most of the children reported previous admissions to a pediatric ICU (63%) or a hospital (82%) for asthma. Despite this, only 48% of the children were taking inhaled corticosteroids before admission. Only 28% of the children reported being followed by an asthma specialist, but, of these, 97% were taking corticosteroids. There were no racial/ethnic disparities in medication use, treatment, or outcomes. CONCLUSION: We found no racial/ethnic disparities in inpatient/outpatient medication usage, treatment, or outcomes between African American and European American children in our cohort. Recurrent admissions to the ICU among children with severe asthma are common, and inhaled corticosteroids usage is relatively low. Asthmatic children with ICU admissions should be followed and treated aggressively by an asthma specialist. PMID- 19998641 TI - The superobese mother and ethnic disparities in preterm birth. AB - OBJECTIVES: We assessed the association between preterm birth and obesity subtypes across racial/ethnic groups. METHODS: We analyzed data on 540981 women from birth cohort files for the State of Florida from 2004 to 2007. Obese women were categorized using body mass index (BMI) values as class I obese (30.0 < or = BMI < or = 34.9), class II obese (35.0 < or = BMI < or = 39.9), class III or extremely obese (40 < or = BMI < or = 49.9), and superobese (BMI > or = 50.0). Logistic regression was used to obtain adjusted estimates. RESULTS: About 28% of women were obese, with the highest rate (40.9%) registered among black gravidas, while whites and Hispanics had comparable rates (24.3% vs 25.5%, respectively). Superobesity was also most prevalent in blacks (1.3%). Among obese women, the risk for preterm birth was greatest among blacks (OR, 1.71; 95% CI, 1.65-1.77), while whites (OR, 1.15; 95% CI, 1.12-1.19) and Hispanics (OR, 1.22; 95% CI, 1.18 1.27) had significantly lower and comparable risk levels. CONCLUSIONS: Extremely obese and superobese women are emerging high-risk groups for adverse birth outcomes, and black women appear to bear the heaviest burden. The disproportionately rising trend in extreme forms of obesity among black women is of utmost concern and represents a clarion call for infusion of more resources into obesity prevention programs in black communities. PMID- 19998643 TI - Disparities in antiretroviral prescribing for blacks and whites in the United States. AB - PURPOSE: The health benefits for blacks on highly active antiretroviral therapy (HAART) lags behind that of whites. We therefore investigated whether this discrepancy in health outcomes is attributable to disparities in the receipt of appropriate HAART between black and white human immunodeficiency virus (HIV) patients. METHODS: The 2000-2005 National Ambulatory Medical Care Surveys were used to identify patients receiving antiretrovirals. Regimens were evaluated for appropriateness according to national antiretroviral guideline recommendations. A multivariable logistic regression model was used to assess black race as a predictor for appropriate HAART. RESULTS: An estimated 4.1 million HIV-related visits with antiretroviral therapy were identified. Eighty-six percent of visits were associated with appropriate therapy; inappropriate therapy was often due to antiretroviral monotherapy. Interestingly, blacks were more likely to receive appropriate therapy in comparison to white patients (94% vs 83%, P < .001). Multivariable analysis revealed black race as an independent predictor for an appropriate regimen (chi2 likelihood ratio, 12.3, P < .001) when controlling for age, gender, insurance status, and geographic region. CONCLUSIONS: Health outcome disparities between black and white HIV patients do not appear to be attributable to differences in receipt of appropriate HAART. Further investigations are warranted to identify factors responsible for these outcome disparities. PMID- 19998642 TI - Racial disparities in revascularization rates among patients with similar insurance coverage. AB - BACKGROUND: Racial disparities in coronary revascularization--percutaneous coronary intervention (PCI) and coronary artery bypass grafting (CABG)--have been extensively documented. However, it is unclear whether disparities are consistent among patients with similar health insurance coverage. Our objective was to assess racial disparities in coronary revascularization among white, black, and Hispanic patients with similar insurance coverage hospitalized with acute myocardial infarction (AMI). METHODS: We used 2000-2005 state inpatient data for 9 states to identify white, black, and Hispanic patients hospitalized with AMI. Patients were grouped into 3 health insurance cohorts: (1) Medicare, (2) private insurance, and (3) Medicaid/uninsured. We examined use of revascularization (PCI or CABG) among blacks and Hispanics as compared to whites in each of the 3 insurance cohorts. RESULTS: The 418 study hospitals admitted 430509 AMI patients with Medicare, 238956 with private insurance, and 74926 patients who were uninsured/Medicaid. In unadjusted analyses, black and Hispanic patients were significantly less likely to receive in-hospital revascularization among the Medicare cohort (38.9% vs 44.9% vs 47.3%, P < .001), privately insured cohort (62.9% vs 69.7% vs 74.2%, P < .001), and uninsured/Medicaid cohort (55.2% vs 61.0% vs 68.4%, P <.001). In Cox models adjusting for patient demographics, comorbidity, and clustering of patients within hospitals, blacks were approximately 25% less likely and Hispanics 5% less likely to receive revascularization as compared to whites with similar insurance. CONCLUSIONS: Blacks hospitalized with AMI are significantly less likely to receive revascularization when compared to whites and Hispanics with similar health insurance. Our data suggest that patients' ability to pay for costly procedures is unlikely to explain racial disparities. PMID- 19998644 TI - Trend and racial disparities in infant mortality rate in Texas from 1990 to 2004. AB - PURPOSE: To determine the trend and disparities in infant mortality rate (IMR) from 1990 to 2004 in Texas, which is one of the few states with severe physician shortage in the United States. METHODS: Trend analysis and Poisson regression were performed with data from Texas Department of State for Health Services and other Web sites. RESULTS: IMR decreased from 1990 to 2000 followed by a trend of increase from 2000 to 2004 in Texas. The recent trend of increase in IMR was observed in all ethnic groups. IMR was associated with ethnicity, area of residence in Texas (rural vs urban areas in east vs south vs west), median income index, and primary care physician supply (primary care physician to population ratio). IMR in blacks was more than 2 times that of other ethnic groups, and the gap had been increasing since 1997. The recent increasing trend in IMR coincided with a decreasing trend in primary care physician supply and a decrease or a slower increase in median income index. CONCLUSIONS: IMR increased in recent years, particularly in African Americans in Texas. Measures should be taken to reverse the worsening trend in IMR and to reduce regional and racial disparities. PMID- 19998645 TI - A peer-to-peer campaign to promote organ donation among racially diverse college students in New York City. AB - PURPOSE: Due to the documented disparities in organ transplantation, individuals from racially diverse backgrounds are encouraged to register as donors and communicate their donation intentions to family. The present study reports an intervention aimed at addressing students' fears and misconceptions related to organ transplantation on college campuses with racially diverse student enrollments. The intervention uses peer-to-peer campaigns at colleges in New York City in an attempt to increase declarations of intent to donate and to educate students about transplantation. PROCEDURES: Six campuses in the New York City area participated in the intervention. Students participating in the college campaign intervention were educated about donation through active learning methods and implemented campus-wide campaigns to recruit fellow students as donors. RESULTS: Student campaigners reported increases in topic salience and self-efficacy from precampaign to postcampaign activities. In addition, rates of donor registration and family communication improved over time. Across 6 campuses, campaigners were able to recruit 1019 students as registered donors. CONCLUSIONS: Participation in the campaign course increases students' vested interest in donation and has proven effective at increasing donor registration among racially diverse groups. PMID- 19998646 TI - A pilot study to explore knowledge, attitudes, and beliefs about sickle cell trait and disease. AB - INTRODUCTION: In the United States, newborn screening programs universally identify newborns with sickle cell disease (SCD) and heterozygote carriers (sickle cell trait [SCT]). Although there is a consensus to disclose SCT to parents, there are limited empirical data about whether and how this information is transmitted to the carrier children. METHODS: In-person questionnaires were administered to parents with SCT and parents of a child with either SCD or SCT to examine the knowledge, attitudes, beliefs, and disclosure patterns about SCT of parents. RESULTS: Fifty-three adults were interviewed, half (27) of whom had a child with SCD. There was significant misunderstanding about sickle cell inheritance (mean score, 68%), but parents who have a child with SCD have better knowledge compared to those without a child with SCD (78% vs 58%, p = .002). Respondents perceive minimal stigma associated with SCT. Unless there is an affected proband, individuals with SCT rarely receive counseling or education outside of the family. CONCLUSIONS: There is significant misinformation about what it means to be a carrier and its health and reproductive implications. Formal professional counseling is rare, especially for those families without an affected proband. Strategies to increase the utilization of counseling and improve genetic literacy are necessary. PMID- 19998647 TI - In the Journal of the National Medical Association 100 years ago: salutatory. PMID- 19998648 TI - Social immunization: a public health approach for the management of substance abuse. PMID- 19998649 TI - Mack Roach III, MD, expert radiation oncologist and prostate cancer specialist, par excellence! AB - Mack Roach, III, MD, is a son of Texas who was born in the east Texas town of Palestine. Dr Roach recalled that while growing up, he, along with his 4 brothers, was strongly and lovingly supported by their parents, especially their mother, who was, as Dr Roach recalled, "always there" for them. He went on to remark that his father taught him 2 important life lessons. One lesson his dad imparted to him was that "whatever you do, be the best," and equally as important, his dad taught him how to play dominos. Following his early youth in Texas, he and his family, relocated to northern California to the San Francisco Bay Area. After completing high school there, he entered Morehouse College in Atlanta, Georgia, where he completed his undergraduate studies in physics, finishing with honors in 1975. He later attended medical school at Stanford University in California, graduating in 1979. Dr Roach is married and has 2 daughters: Imani, who is in a PhD program at Harvard University, and Sarah, who is currently in high school. Dr Roach is currently a tenured professor of radiation oncology and urology, and serves as the chairman of the Radiation Oncology Department at the University of California Medical Center in San Francisco, California. He has received many accolades in his career to date such as being voted one of the Best Doctors in the Bay Area (1997, 1998) and Best Doctor in America in (1996, 1997, 2003, and 2005), and he was awarded the Health and Wellness Award from the 100 Black Men of the Bay Area (1999). He has traveled the world lecturing on the intricacies of prostate cancer treatment from a radiotherapeutic perspective, both technical and clinical, including a United Nations meeting on prostate cancer in Vienna, Switzerland. PMID- 19998650 TI - The price of soft drinks. PMID- 19998651 TI - What is trust worth? PMID- 19998652 TI - Dentistry and medicine--a new crossing point. PMID- 19998653 TI - Destructive periodontal diseases, systemic inflammation, and atherosclerotic complications: the emerging role of the dental profession. AB - An emerging body of evidence has associated moderate to severe periodontitis with atherosclerotic complications. The contribution of periodontitis to systemic inflammation may account for this association in view of the pivotal role inflammation plays in atherosclerotic complications. Periodontal therapy has been shown to decrease systemic inflammation and to improve early atherosclerotic events; however, to date, periodontal therapy has not been shown to decrease subsequent atherosclerotic complications although aggressive treatment in at-risk populations appears warranted. PMID- 19998654 TI - Anti-microbial peptides for plaque control and beyond. AB - Anti-microbial peptides perform many functions in the oral cavity. They may provide protection against microbial pathogens, assist in oral biofilm control, and function as an important part of the innate immune system in response to local and systemic infection. Synthetic versions of these peptides may be useful to supplement natural anti-microbial peptides or as therapeutic agents. PMID- 19998656 TI - Craniofacial tissue regeneration: where are we? AB - This paper provides a brief review of adult stem cells and their potential clinical applications, specifically in craniofacial regeneration. The initial discovery of stem cells from a variety of tissues has infused tremendous research, and, in conjunction with bioengineering technologies, has potential to transform clinical dentistry. PMID- 19998657 TI - Treating the older adult dental patient: what are the issues of concern? AB - Just as aging successfully requires a multifaceted approach that includes full engagement in life, maintenance of high physical and cognitive function, and avoidance of disease and related disability, so does the care of adult patients. This geriatric treatment model suggests that understanding the psychosocial, behavioral, and medical presentation of the older patient may prove to be the key to the ultimate success of the dental/oral treatment arrived at collaboratively by the dentist and the older patient. PMID- 19998655 TI - Improving oral cancer survival: the role of dental providers. AB - Oral cancer accounts for 2 percent to 4 percent of all cancers diagnosed each year in the United States. In contrast to other cancers, the overall U.S. survival rate from oral cancer has not improved during the past 50 years, mostly due to late-stage diagnosis. Several noninvasive oral cancer detection techniques that emerged in the past decade will be discussed, with a brief overview of most common oral cancer chemopreventive agents. PMID- 19998659 TI - Leap of faith. PMID- 19998658 TI - Oral health and HIV infection: a chronic disease model. AB - HIV disease is now considered a chronic illness requiring continued management and monitoring. However, for those with poor access to anti-retroviral medications, the disease continues to be associated with higher morbidity and mortality. With the expansion of the HIV pandemic into vulnerable subpopulations, HIV care requires coordinated and integrated care for a complex mix of psychosocial and clinical services that must include oral health care. PMID- 19998660 TI - Antibiotic prophylaxis in dentistry and oral surgery: use and misuse. AB - INTRODUCTION: Antibiotic prophylaxis (AP) still represents a common but often misused procedure in dental practice, thus aggravating the risk for antimicrobial resistance and adverse effects occurrence. AIMS AND METHODS: Our primary objective is to review the available scientific evidence regarding AP in dentistry both among healthy subjects and medically compromised patients. Additionally, the latest available guidelines provided by some of the most authoritative associations are here discussed. RESULTS AND CONCLUSIONS: AP is advisable only in a small percentage of patients where a risk of severe infective complications (i.e. infective endocarditis and prosthetic joint infection, septicaemia in severely immuno-compromised patients, bisphosphonate-related osteonecrosis of the jaw) exists. On the contrary, little or no scientific evidence exists for AP in subjects with other systemic diseases as well as in healthy individuals. This pioneering recommendation is strongly evidence-based, since a consistent association between any dental procedure and the development of local and distant infective complications is still lacking. In addition, the daily bacteraemia secondary to routine habits has been thought to be robustly associated with a greater risk of systemic disease bacterial-related than a single dental procedure exposure. Compliance of general dental or family practitioners to the current recommendations seems not to be optimal, thus, efforts to improve it should be planned and undertaken. PMID- 19998661 TI - Sample infection control needs assessment survey data from eight countries. AB - OBJECTIVES: To compare knowledge, attitudes and practice of standard methods in infection control by dentists in eight countries using a standardised questionnaire. METHODS: Self-administered questionnaires on dental infection control and safety (IC&S) knowledge, attitudes and practice were distributed to a convenience sample of dentists in India, Pakistan, Thailand, Philippines, Taiwan, China, South Korea and the United States of America. RESULTS: Knowledge and practice of infection control among the Asian dentists lagged behind the levels of US dentists. Availability and affordability of equipment and materials were some of the reasons for the practice of infection control and safety. Attitude towards universal/standard precautions in controlling bloodborne pathogens was comparable between groups surveyed. Tested knowledge and practice of infection control was acceptable but the attitude and understanding of universal/standard precautions was ambiguous among both Asian and USA dental practitioners. Education in dental infection control should focus on improving the attitudes of practitioners towards universal precautions. CONCLUSIONS: Knowledge and practice of dental IC&S was high among the USA respondents. Immunisation and IC & S should be stressed in Asia. Information obtained through this study could be used to plan and implement policies in improving dental infection control and occupational safety in individual countries. PMID- 19998662 TI - A comparative study of the health care systems of Canada and Saudi Arabia: lessons and insights. AB - INTRODUCTION: Understanding the structure of a health care system is essential in improving public health policies and health outcomes. OBJECTIVES: To describe and compare the health care systems of Canada and Saudi Arabia; to discuss possible lessons that could be learned from both for policy-making purposes. METHODS: A comprehensive method was used to describe the national health care systems of both countries. For each country, the system is described by: context, ownership, delivery, financing, organisational structure, target groups, and comprehensiveness of services. RESULTS: In Canada, the Medicare system provides comprehensive medical services except for dental, optometric, chiropractic, pharmacologic and home care services. The dental care system is financed privately (94%) and is owned and delivered by private for-profit dental practitioners. In Saudi Arabia, the government sector is owned, delivered, and financed by the government and provides free comprehensive medical and dental services. The same services are provided by the private sector, but under governmental supervision. Among the relevant lessons: access to care, accountability, quality assurance, mix and reimbursement of providers. CONCLUSIONS: Canada can learn about different approaches to socialising the dental care system. Saudi Arabia can improve the implementation of quality assurance practices and management. PMID- 19998663 TI - A silica toothpaste containing blue covarine: a new technological breakthrough in whitening. AB - There continues to be a demand from consumers and patients for products that whiten teeth. These products are generally either peroxide-based bleaching formats to improve intrinsic tooth colour or whitening toothpastes with enhanced physical and chemical cleaning properties to remove and prevent extrinsic tooth stain. Through the understanding of colour science and its application to tooth colour a new approach to tooth whitening is now possible from toothpaste. This is achieved by the deposition of blue covarine onto the tooth surface where it changes the optical properties of the teeth such that their appearance is both measurably and perceivably whiter immediately after treatment. In vitro and in vivo studies confirm the efficacy of this whitening approach in a silica based whitening toothpaste containing blue covarine. In addition, this toothpaste has been shown not to have an undue degree of abrasivity to enamel or dentine compared to other relevant commercially available products, and is an efficacious source of fluoride. PMID- 19998664 TI - Strategies to promote better access to over the counter products for oral health in Europe: a Delphi survey. AB - OBJECTIVES: 'Over The Counter' (OTC) is a pharmaceutical product or medicine whose distribution or the administration does not require medical authorisation, and which can be used by consumers on their own initiative. This can be to prevent, relieve or treat symptoms or moderate pathologies and whose use, under the forms, packaging and authorised formulation are safe for the consumer. A Delphi consultation in the perspective of coming to a consensus was initiated to identify the current and future orientations on the best strategies of administration of OTC products for oral health in Europe. METHODS: A Delphi Survey was conducted with 54 experts from 23 countries in Europe. Each indicator statement was considered to be in consensus if the expert's opinion rating was of 4 or 5 for more than 75% in a scale of seven categories. The first questionnaire concerned self medication and the situation of OTC prescriptions in 2006. The second included 19 statements focused on the possible role of OTC products in dental practice. Both qualitative and quantitative analyses were created. RESULTS: There was a strong consensus that the population's common practices towards OTCs should be modified. Practitioners should possess communication qualities allowing them to share their power and to advise patients of their decision-making concerning oral care. CONCLUSIONS: The Delphi Survey was successful in underlining that dentists have to be involved in oral health OTC prescription which, currently, seems unsatisfactory. OTC products and especially fluoride toothpaste improve oral health but have to be available, accessible and used advisedly. PMID- 19998665 TI - Systemic and local applications of steroids in endodontics: an update review. AB - Up to 80% of endodontic patients who report with preoperative pain continue to experience some level of pain following the endodontic procedure. Various classes of drugs have been studied for the management of post-treatment endodontic pain. Since endodontic pain is often associated with chronic inflammation, the presence of bacterial by-products, influx of primed immune cells and activation of the cytokine network and other inflammatory mediators, pain may be reduced by administration of glucocorticoid steroids. The aim of this paper is to review the pharmacology and mechanisms of actions of steroids as well as their indications for endodontics, contraindications, dosages and side-effects. PMID- 19998668 TI - Doing well while doing good. PMID- 19998666 TI - Modulation of the host inflammatory response in periodontal disease management: exciting new directions. AB - BACKGROUND: New strategies for periodontal disease management have been emerging as more is learned about the role of the host response. Our increasing understanding of inflammation and its resolution has opened the door to the study of new periodontal treatment strategies. This review examines periodontal disease in the light of a new understanding of the role of inflammation in disease expression thus setting the stage for the development of new prevention and treatment strategies of a widespread disease. METHODS: We examined current publications and focused on articles relating to anti-inflammatory and pro resolution mechanisms in periodontal disease. RESULTS: Recent research has examined the inflammatory and resolution cascade in greater detail while looking at endogenous and exogenous mediators that can be utilised to achieve therapeutic end-points. The possible introduction of 'resolution indices' for drug testing warrants a new look at pharmacologic agents that might have been overlooked for their beneficial effects in periodontal disease treatment. CONCLUSION: The emerging awareness of inflammation and its control in periodontal disease management underscores the importance of exploring inflammatory pathways and mediators, thus exploring new ways to control inflammation. This direction of research promises a new era in drug discovery and therapeutics for periodontal disease treatment. PMID- 19998669 TI - The future of documentation. PMID- 19998670 TI - Frequent episodes of fainting. PMID- 19998671 TI - Patient information. Bladder outlet obstruction. PMID- 19998672 TI - Low libido in postmenopausal women. PMID- 19998673 TI - Type 1 diabetes in children and adolescents. An overview of real-life issues. PMID- 19998674 TI - Dysphagia in older adults. PMID- 19998675 TI - Through the looking glass. The multiple faces of prescriptive authority. PMID- 19998676 TI - Comparing NPs, PAs and physicians. PMID- 19998677 TI - Both sides of pharmaceutical promotion. PMID- 19998678 TI - Resolving work conflicts. A little forethought is most effective. PMID- 19998679 TI - Relationships. PMID- 19998680 TI - Speed of processing in older adults: a cognitive overview for nursing. AB - With advancing age, subtle declines in various cognitive abilities occur. One ability that is particularly important as people age is speed of processing. Speed of processing is suggested to impact other cognitive abilities; in fact, some studies suggest that other cognitive abilities depend on speed of processing as a foundational cognitive resource. Studies also indicate that declines in speed of processing influence everyday functioning and driving behavior. Fortunately, cognitive remediation therapy designed to improve speed of processing has been shown to accentuate speed of processing and everyday functioning in older adults. Nurses are in a unique position to listen to patient's cognitive complaints and can offer suggestions and referrals to help older adults maintain this cognitive ability. Implications for nursing research are posited. PMID- 19998681 TI - Sex differences in depressive symptoms and their correlates after mild-to moderate traumatic brain injury. AB - The purpose of this secondary data analysis, guided by allostatic load theory, was to compare depressive symptoms and their correlates in men and women following mild or moderate traumatic brain injury (n = 159). Using general linear modeling procedures in the Statistical Analysis Software, women reported significantly higher Center for Epidemiological Studies-Depression scores compared with men. According to the Neurobehavioral Functioning inventory subscales, women also reported higher somatic and motor symptoms and difficulties with memory and cognition. Further, women within the first 6 months of their injury reported higher levels of depressive and depressive-somatic symptoms, perceived chronic stress, pain, memory difficulties, and somatic symptoms. These findings were no longer present at the 6- to 12-month or >12-month cutoffs. Women's depressive symptoms during the early recovery period are explained by higher symptom loads and perceived stress, yet mechanisms responsible for these differences remain to be elucidated. Future research is needed to describe hormonal, perceptual, or brain structure differences that may account for these findings. Findings from such research will most likely to contribute to our understanding of postconcussion syndrome. PMID- 19998682 TI - Content and characteristics of goals created during a self-management intervention for people with epilepsy. AB - Goals are presented in the chronic illness literature as effective strategies to help people adopt self-management behaviors; however, not much is known about the types and characteristics of individuals' goals. The purpose of this study was to examine goal setting among people with epilepsy who participated in the WebEase program. WebEase is an Internet-based, theory-driven, self-management program with modules on medication adherence, stress management, and sleep habits. Participants had the opportunity to create and evaluate goals over the course of 6 weeks, with 2 weeks for each module. The goals were analyzed using three dimensions: content, specificity, and proximity. Most participants in the sample wrote goals for each week of the program. Several main content areas emerged within the modules. Goal quality, measured by specificity and proximity, did not differ according to readiness for behavior change. Readiness to change did not differ between those who wrote a goal and those who did not. The diversity of goal content and quality indicates that individuals should be supported in goal development and encouraged to set their own self-management goals, regardless of their readiness for behavior change. PMID- 19998683 TI - The impact of pain and other symptoms on quality of life in women with relapsing remitting multiple sclerosis. AB - The purpose of this study was to assess pain, fatigue, depression, sleep disturbance, and quality of life (QOL) in women with relapsing-remitting multiple sclerosis (RRMS) compared with healthy controls. A prospective, cross-sectional, matched-control study was conducted in women with RRMS compared with healthy women. Compared with healthy women, women with RRMS had (a) greater pain presence over 7 days (67%), (b) higher pain intensity, and (c) more pain interference. Pain had a negative impact on fatigue, depression, and sleep in both groups. In all participants, fatigue, depression, and sleep disturbance contributed to decreased mental QOL (mental component summary of QOL scores). Pain has significant nursing implications for women with RRMS. Pain often occurs in association with fatigue, depression, and sleep disturbance, which can lead to a decreased mental QOL. PMID- 19998684 TI - Nurse practitioner-based sign-out system to facilitate patient communication on a neurosurgical service: a pilot study with recommendations. AB - Failure to communicate important patient information between physicians causes medical errors and adverse patient events. On-call neurosurgery physicians at the Toronto Western Hospital do not know the medical details of all the patients that they are covering at night because they do not care for the entire service of patients during the day. Because there is no formal handover system to transfer patient information to the on-call physician, a nurse practitioner-based sign-out system was recently introduced. Its effectiveness for communication was evaluated with preintervention-postintervention questionnaires and by recording daily logins. There was a statistically significant decrease in number of logins after 8 weeks of use (p = .05, Fisher's exact test), and the tool was abandoned after 16 weeks. Modifications identified to improve the system include the ability to sort by attending physician and to automatically populate the list with new patients. Effective communication is important for reducing medical errors, and perhaps these modifications will facilitate this important endeavor. PMID- 19998685 TI - Stroke education for nurses through a technology-enabled program. AB - Today's nurse faces many challenges in the workplace. Required to keep up in a constantly changing knowledge-based environment, he or she must balance complex professional responsibilities, staffing shortages, and increased acuity among the patient population. Continuing education must, therefore, be highly flexible and responsive to the personal and professional needs of the nurse learner. Technology-supported continuing education is suggested to be an appropriate way of meeting the learning needs of busy working nurses. The Stroke Best Practices for Nursing project used three complementary and integrated educational technologies-a-Web-based learning site, Web casting (live and archived), and two way interactive videoconferencing--to deliver a minicourse focused on best practice stroke care to nurses working in northeastern and northwestern Ontario, a geographical area of approximately 600 km. In total, 96 nurses participated in the educational part of the program; 46 of the 96 (47%) took part in the assessment of the program. On the basis of this assessment strategy and the nurses' requests for other programs that do not use traditional face-to-face classrooms and lecture, the value of using educational technologies in health based continuing education was strongly identified. This article describes key components of the project and celebrates the partnership among the organizing stakeholders: faculty in the school of nursing at the Laurentian University, the West Greater Toronto Area Stroke Network, and the Ontario Telemedicine Network. The article further describes findings related to the program's impact on participants' perceptions of competence as caregivers for stroke patients, participants' confidence using technology for educational purposes, and participants' satisfaction with the overall program. PMID- 19998686 TI - Research and quality improvement activities: when is institutional review board review needed? PMID- 19998687 TI - Pharmacological management for the adult migraine sufferer. AB - Migraine is like any other chronic illness. Patients require education, reassurance, and support from their healthcare providers. An accurate history is imperative to successful treatment. Maintaining a detailed headache diary is important for the patient as well as the healthcare provider to help sort out triggers and possible errors in how patients take their medications. Choosing the most appropriate abortive agent and, if needed, preventive treatment leads to successful migraine management. PMID- 19998688 TI - Promoting your voice and vision with Canadian Nurse. PMID- 19998689 TI - Off-label prescribing of the nicotine patch for psychiatric patients. PMID- 19998690 TI - Setting a new standard of care in nursing homes. AB - The Winnipeg Regional Health Authority's introduction of a full-time nurse practitioner in a 116-bed non-profit nursing home provided an opportunity to explore a collaborative relationship between an NP acting as the primary care provider and a single physician serving as the consultant for complex care and after-hours care. The outcomes were measured in terms of resident and family satisfaction, quality of care indicators and cost effectiveness. Data were collected from pre-existing quality indicators, including a resident/family satisfaction survey, transfers to acute care, and medication use statistics. Unstructured interviews were also conducted with nursing staff and members of the interdisciplinary team. Dramatic improvements in medication use were observed, including a 17 per cent reduction in overall drug costs, a 55 per cent decrease in polypharmacy rates and a 63 per cent reduction in antipsychotic drug use. Transfers to emergency decreased by 20 per cent. Family satisfaction with the quality of health care provided to residents increased by 24 per cent. The collaborative practice of an NP with physician consultation is an effective way of delivering quality care to nursing home residents. PMID- 19998691 TI - Challenge, opportunity and adventure: stories from Newfoundland and Labrador. Interview by Tara Tosh Kennedy. PMID- 19998692 TI - Mental health nursing in Afghanistan. PMID- 19998693 TI - A catastrophe in health care. PMID- 19998694 TI - Domestic violence screening in pregnancy. AB - PURPOSE: Domestic violence is an important health concern that has been shown to have adverse effects on maternal and neonatal outcomes. The objectives of this study were to compare the prevalence of prenatal screening for domestic violence in a hospital-based resident clinic setting with screening practices in private obstetric offices in Honolulu, Hawai'i and to explore physician attitudes towards domestic violence screening during pregnancy. METHODS: A retrospective chart review was conducted at Queen's Medical Center in Honolulu, Hawai'i in women who delivered between 2003 and 2004. A 6 item written survey was also given to all attending and resident physicians with obstetric privileges. Descriptive statistics including frequency measures were generated and chi square tests were used to compare categorical variables. RESULTS: A total of 270 charts were reviewed. There was a statistically significant difference (p < 0.01) between the number of antepartum patients from the resident clinic (2.4 percent) and antepartum patients from private obstetric practices (39.3 percent) that were screened for domestic violence. While the majority of respondents (77.6%) to the domestic violence survey were aware that the American College of Obstetricians and Gynecologists recommends domestic violence screening in pregnancy most respondents (69.0 percent) indicated that they "never or rarely" screened their patients for domestic violence. CONCLUSION: Despite professional recommendations and an awareness of these recommendations, between 2003 and 2004, routine prenatal screening for domestic violence was markedly lacking for patients in this study population. PMID- 19998695 TI - "Reversible" late bioprosthetic aortic valve stenosis with spontaneous recovery. AB - BACKGROUND: Advantages of bioprosthetic valve replacement in patients with normal sinus rhythm are avoidance of the need for long-term anticoagulation and reduced hemorrhagic accidents. On the other hand, low durability of the valve and a higher re-operation rate are known disadvantages. Bioprosthetic valve thrombosis and related embolism are considered to be rare complications. CASE REPORT: This 80-year-old man underwent aortic valve replacement with a 23 mm porcine prosthesis and concomitant single vessel right coronary artery bypass graft with an autogenous vein. Two years after the initial surgery the patient developed progressive dyspnea. Continuous wave doppler echocardiography showed a greatly elevated mean pressure gradient of 48 mmHg and an aortic valve area of 0.45 cm2 compatible with severe aortic stenosis. Two months later prior to the elective repeat aortic valve replacement surgery, a transthoracic echocardiography showed a significantly reduced mean pressure gradient of 19 mmHg and the effective valve area was 1.3 cm2, and the surgery was cancelled. CONCLUSION: The authors report a case of "reversible" late porcine aortic valve stenosis with spontaneous resolution, likely due to thrombus formation on the valve leaflets. Spontaneous thrombosis of the Medtronic mosaic porcine aortic bioprosthetic valve may occur in the absence of any identifiable causes. PMID- 19998696 TI - Problem based learning and academic performance in residency. AB - BACKGROUND: In the Problem Based Learning (PBL) curriculum, knowledge is acquired by solving medical cases. Because of this quality, it has been hypothesized that residents trained in a PBL curriculum during medical school will be better equipped to succeed during residency. The purpose of this study was to compare the academic performance of obstetrics and gynecology residents who were trained using a PBL curriculum during medical school with those who were trained in a traditional curriculum. METHODS: A retrospective analysis was performed on residents at the University of Hawai'i Obstetrics and Gynecology Residency Program between 1997 and 2007. Scores on standardized tests were compared between residents who attended a PBL medical school (PBL group) and residents who attended a traditional medical school (non-PBL group) using a t-test analysis. United States MedicalLicensing Examination (USMLE) Step 1, USMLE Step2, and the Council on Resident Education in Obstetrics and Gynecology (CREOG) examination scores for all four years of residency were compared between the two groups. RESULTS: Thirty-five students had complete data. There were no significant differences between the mean scores in PBL group versus the non-PBL group for the Step 1 (205.7 vs. 202.0, p = 0.491), CREOG PGY1 (197.8 vs. 195.7, p = 0.711), CREOG PGY2 (202.7 vs. 198.1, p = 0.394), CREOG PGY3 (197.3 vs. 201.1, p = 0.545), and CREOG PGY4 (202.4 vs. 198.8, p = 0.531). However, there was a significant difference between the mean scores of the two study groups for the USMLE Step 2 (215.1 vs.202.1, p = 0.046). CONCLUSIONS: Although PBL students performed significantly better on the USMLE Step 2, the academic performance of obstetrics and gynecology residents who attended a PBL medical school and those who attended a traditional medical school was similar. PMID- 19998697 TI - A case with transient anterolateral wall ballooning syndrome; new variant form of Takotsubo cardiomyopathy? AB - Proposed to be a new variant form of the left ventricular apical ballooning syndrome or the so-called "Takotsubo cardiomyopathy" the presented case has several clinical characteristics resembling previous reported cases of this particular syndrome except for its unique anterolateral wall akinesis and sparing basal and apical wall motion with preserved left ventricular ejection fraction. PMID- 19998698 TI - Simulation crisis team training effect on rural hospital safety climate (SimCritter). PMID- 19998699 TI - The physician's role in health and social justice. PMID- 19998700 TI - Stopping cancer cells in their tracks... PMID- 19998701 TI - CNA negotiates alternate plan for safe prescribing--APNs moving away from collaborative agreements. PMID- 19998702 TI - Colorado Nurses Association advocates for APN scope of practice. SB 007 postponed indefinitely, then resurrected. PMID- 19998703 TI - Porter Hospital achieves Magnet designation-nursing reflects on meaning. PMID- 19998704 TI - Forensic nursing: evolving practice in response to the epidemic of violence. PMID- 19998705 TI - 'Sled docs' and quest for adventure in Alaska. PMID- 19998706 TI - Malpractice relief. Lower premiums, tort reform add to Alaska's appeal. PMID- 19998707 TI - The medical school applicant pool from Alaska: implications for the development of medical education in the state. AB - Several recent physician workforce reports in Alaska have called for a rapid increase in the number of state-supported medical school positions to between 30 and 50 new students per year, preferably through WWAMI - Alaska's Medical School. Here we compare applicant and matriculant data for Alaska WWAMI and for Alaskans applying to all U.S. allopathic medical schools to national applicant and matriculant trends gathered from databases of the Association of American Medical Colleges. The assessment demonstrates that 1) changes in the number of Alaska WWAMI applicants parallels changes in the number of Alaskans applying to all U.S. medical schools, but these do not track changes in the national applicant pool, 2) historical records suggest that a fundamental change is needed to provide an applicant pool to support a class size of 50 within the next decade, 3) smaller states with independent, four-year medical schools generate more medical school applicants per capita than Alaska. PMID- 19998708 TI - Family violence. Prevention doesn't have to be an impossible dream. PMID- 19998709 TI - Sled docs. Physicians & the Iditarod: Dr. Jim Lanier. PMID- 19998710 TI - Training and racing. PMID- 19998711 TI - Sled docs. Physicians & the Iditarod: Dr. Bob Bundtzen. PMID- 19998712 TI - Sled Docs. Physicians & the Iditarod: Dr. Lily Lou. PMID- 19998713 TI - Rates and correlates of postpartum depression in Alaska. PMID- 19998714 TI - Management of the open abdomen: from initial operation to definitive closure. AB - The open abdomen is a relatively new and increasingly common strategy for the management of abdominal emergencies in both trauma and general surgery. The use of an abbreviated laparotomy can reduce mortality associated with conditions such as abdominal compartment syndrome; however, the resulting open abdomen is a complex clinical problem. Modern techniques and technologies are now available that allow for improved management of the open abdomen and the progressive reduction of the fascial defect. Indeed, recent evidence indicates that a large proportion of patients treated with open abdomen can now be closed within the initial hospitalization. These techniques and technologies include the appropriate use of negative pressure therapy and synthetic or biologic repair materials. It is essential that general and trauma surgeons understand the core principles underlying the need for and management of the open abdomen. Toward this goal, an Open Abdomen Advisory Panel was established to identify core principles in the management of the open abdomen and to develop a set of recommendations based on the best available evidence. This review presents the principles and recommendations identified by the Open Abdomen Advisory Panel and provides brief case studies for the illustration of these concepts. PMID- 19998715 TI - [MicroRNAs in a variety of life phenomena]. PMID- 19998716 TI - Efficacy of mediodorsal thalamic nucleus stimulation in a rat model of cortical seizure. AB - OBJECTIVE: The mediodorsal thalamic nucleus (MD) has strong connectivity to the limbic systems and frontal lobe. The aim of this study was to determine whether the mediodorsal thalamic nucleus influences seizure induction by electrical stimulation in the rat frontal cortex. MATERIALS AND METHODS: Juvenile male rats (n=7) were used in the experiment. At postnatal day 28 (P28), stainless steel skull screws were inserted for stimulating cortices. Depth-stimulating electrodes were stereotaxically inserted into the MD of the bilateral thalamus. Electrical stimulation, which produces afterdischarges, was applied to the frontal cortices once/day for four consecutive days from postnatal day 38. Additional conditioning electrical stimulation to the bilateral MD was fixed at 0.1 mA, and the stimulus frequency was increased from 0 Hz, 1 Hz, and 5 Hz to 10 Hz. All seven rats were stimulated with those frequencies by rotation every other day. The data of animals with 0 Hz MD stimulation were defined as controls. Afterdischarge thresholds and durations were measured at each frequency. RESULTS: Seizures were accompanied by bilateral tonic-clonic convulsions and cortical spikes. Seizure behaviors were not different among the groups and there were no statistically significant differences in the cortical afterdischarge (AD) thresholds between animals with MD stimulations and controls in all frequencies. In the cortical AD durations, there were also no statistically significant differences between animals with MD stimulation and controls although the mean duration at 10-Hz stimulation was less than the control animals. CONCLUSION: These results indicated that thalamic MD stimulation might not suppress epileptic seizure induction. Further studies are needed to analyze other stimulus parameters, altered stimulus locations, and different test paradigms. PMID- 19998717 TI - A Japanese case of familial Mediterranean fever with a MEFV gene mutation. AB - A 17-year-old Japanese male with a three-year history of recurrent episodic chest pain and fever, each lasting for one to three days, was diagnosed with familial Mediterranean fever (FMF) because of the following: 1) short attacks of chest pain and fever recurring at varying intervals; 2) no symptoms with a sense of well-being between attacks; and 3) identification of the Mediterranean fever gene (MEFV) mutation demonstrating M6941. Although FMF has been described primarily in several limited ethnic groups, a limited number of cases have been reported in Japan. No specific diagnostic tests are commercially available for FMF so identifying the characteristic clinical picture of FMF is important. PMID- 19998718 TI - Metabolic changes during cataract formation by ultraviolet radiation in the incubated rabbit lens. AB - We studied the energy metabolism of cataracts induced by ultraviolet (UV) irradiation by observing metabolic changes in lenses using 31P-, 1H-, and 13C nuclear magnetic resonance (NMR) spectroscopy. % of hexose monophosphate shunt flux activity increased over time. The lactate/glucose ratio in the UV irradiation group decreased to about half of that of the corresponding control group (11.6 +/- 2.0% vs. 20.7 +/- 1.7%, respectively, p < 0.05) after 10 h of irradiation and to about 30% of that of the control group (25 +/- 2% vs. 92 +/- 6%, respectively) after 24 h. The adenosine triphosphate (ATP) level significantly decreased after 3 h of irradiation (86 +/- 29%, p < 0.05) and continuously decreased to 68 +/- 33% (p < 0.01) after 10 h of irradiation and 26 +/- 2% (p < 0.01) after 24 h of irradiation. Conversely, a significant increase in inorganic phosphate (Pi) was observed after 1 h of irradiation (111 +/- 26%, p < 0.05), and the Pi level gradually increased to 140 +/- 28% after 10 h of irradiation (p < 0.01) and 207 +/- 26% after 24 h (p < 0.01). A significant decrease in alpha-glycerophosphate was noted after 24 h (38 +/- 13%, p < 0.01). The ribose-5-phosphate (R-5-P) level gradually increased after irradiation to 128 +/- 13% (p < 0.05) after 10 h and 141 +/- 21% after 24 h (p < 0.01). The results suggest that of these metabolic changes a marked decline in glycolytic production of ATP, which inhibits membrane metabolism, may be an important cataract-inducing factor following UV irradiation. PMID- 19998720 TI - Prolonged intrauterine device leading to abscess formation and sepsis. PMID- 19998719 TI - Practice self exam: critical self appraisal in family medicine. PMID- 19998721 TI - Fine particulate air pollution and cardiovascular disease. PMID- 19998722 TI - A father's and physician's perspective on Down syndrome. PMID- 19998723 TI - Lewis Blackman Hospital Patient Safety Act. PMID- 19998724 TI - The three cultures and health care reform. PMID- 19998725 TI - Decision-making for treatment planning a cantilevered fixed partial denture. AB - Considerable controversy exists in the dental literature regarding cantilevered pontics. This article discusses basic concepts of the cantilever fixed partial denture (CFPD) in which one cantilevered pontic is supported by only one or two abutment teeth. Three primary factors should be considered carefully to optimize the prognosis for a CFPD: abutment selection, control of functional forces, and rigidity/strength of connectors. Abutments should have a root surface area greater than the tooth being replaced and a crown-to-root ratio of 2:3. They also should exhibit minimal mobility and be vital and periodontally sound. Contact on cantilevered pontics should be light in centric position and nonexistent in excursions. CFPDs ideally should be metal or metal-ceramic, and connectors, which are high-stress areas, require bulk for strength. A cantilevered prosthesis may require more consideration and planning than a conventional fixed partial denture, but when kept within the patient's biological limitations and executed properly, can provide a restorative option with many advantages. PMID- 19998726 TI - Clinical evaluation alveolar ridge preservation with a beta-tricalcium phosphate socket graft. AB - PURPOSE: To determine the efficacy of an alloplastic graft material, consisting of a pure-phase beta-tricalcium phosphate (beta-TCP), in the preservation of ridge volume after tooth extraction and before dental implant placement. Histomorphometric analysis was completed on a few samples to determine the percentage of vital bone over a fixed healing period. MATERIALS AND METHODS: Patients requiring tooth extraction and bone regeneration before implant placement were included in this study. Measurements of alveolar width were made at the time of extraction and the time of implant placement. The extraction sites were grafted with a pure-phase beta-TCP and covered with a barrier. Approximately 6 months after surgery, the sites were reentered for implant placement. Cores were taken of the regenerated material for histologic analysis, with a trephine used as the first bur in preparation for some of the osteotomies. Implants were placed according to the manufacturers' recommendations and loaded at the appropriate time. RESULTS: The beta-TCP placed at the time of grafting extraction sockets was well tolerated in all sites with all of the barriers used. There were no incidences of postoperative infection or graft rejection. At the time of implant placement, much of the graft material had resorbed and been converted to vital alveolar bone. The implant recipient sites were dense and supported placement of endosseous dental implants that were fully stable. The width of the extraction sockets was preserved to 91% of the preoperative width. CONCLUSIONS: Extraction socket grafting with the pure-phase beta-TCP tested in this study and covered with either a resorbable collagen or dense polytetrafluoroethylene barrier is a predictable method for preserving alveolar dimensions. The graft material resorbs to a high percentage in the timeframe desired between extraction and dental implant placement, as shown clinically, radiographically, and histologically. In addition, the regenerated material in the socket has enough density to support implant placement with subsequent loading in the 4- to 6-month period used in this study. PMID- 19998727 TI - Tissue-specific, direct-application class II tooth repair: a case report. PMID- 19998728 TI - Etiology and prevention of acid erosion. AB - Acid erosion is the chemical effect of dietary or gastric acids on enamel and dentine. Unlike dental caries, which is a bacterially mediated condition, acid erosion normally is combined with physical forms of attrition and abrasion. The clinical appearance of acid erosion in the early stages is seen characteristically as hollowed-out lesions on occlusal surfaces and on smooth surfaces as a subtle change in the tooth contour. As the condition progresses, the lesions coalesce and form widespread dentin exposure and coronal destruction. Dietary acids that are present in beverages and fruits potentially can cause acid erosion. However, dietary habits involving frequent consumption of acids are believed to be important determinants in the risk of developing clinical signs of acid erosion. Prevention using fluoride toothpastes, dietary modifications, and calcium-based products probably have a significant clinical effect in reducing the risk of developing acid erosion. PMID- 19998730 TI - Removal of Invisalign retention attachments: a new minimally invasive method. AB - Removal of Invisalign resin retention buttons without damaging underlying enamel is a major challenge. To date, the use of tungsten carbide burs is the most common and fastest--yet a risky-ablation method. Stainbuster, a fiber-reinforced resin bur, has been introduced for removal of surface stains and resin remnants from tooth surfaces. This comparative in vitro and in vivo study proved that a combined technique, using multifluted tungsten carbide burs for fast removal of the bulk of resin followed by Stainbuster grinding for gentle removal of the final resin layer, is a safe and minimally invasive procedure for removing composite buttons from enamel. PMID- 19998731 TI - Impressioning. PMID- 19998729 TI - Mechanism of action of a desensitizing fluoride toothpaste delivering calcium and phosphate ingredients in the treatment of dental hypersensitivity. Part II: comparison with a professional treatment for tooth hypersensitivity. AB - Tooth hypersensitivity can occur when gum recession causes exposure of dentin. Tiny tubules, which permeate dentin, provide open passageways from the mouth to the intradental nerve in the pulpal cavity. Under such circumstances, stimuli in the mouth can cause pressure on the intradental nerve, leading to pain. Sealing the outside of the tubules with an impermeable substance can effectively treat hypersensitivity. One such clinically proven composition is a professionally applied tooth desensitizer, which has been shown to initially produce a layer of amorphous calcium phosphate (ACP) on the surface of dentin. Under the influence of fluoride, ACP reforms as hydroxyapatite (HAP), which has essentially the same composition as tooth mineral. Three fluoride toothpastes that deliver calcium and phosphate salts to the teeth also have been demonstrated in clinical trials to relieve hypersensitivity. This study compared the mechanism of action of these toothpastes to that of the professional desensitizer. A single application of the professional desensitizer or multiple applications of any of the three toothpastes was shown to reduce dentin permeability. A conventional fluoride toothpaste also was found to inhibit fluid flow through the dentin but to a lesser degree than the other toothpastes. The desensitizer and the three toothpastes were found to occlude the dentinal tubules with a layer of calcium phosphate that had a calcium-to-phosphate ratio consistent with the formation of ACP or HAP. The morphology of the coherent mineral layer formed by Arm & Hammer Enamel Care Sensitive was similar, especially to that produced by the desensitizer. In contrast, the conventional toothpaste left localized areas of surface residue composed of silica particles. The mechanism of action of the three toothpastes that deliver calcium and phosphate salts is the same as that of the professional desensitizer. PMID- 19998732 TI - While we were arguing. PMID- 19998733 TI - Coagulopathy associated with superwarfarin exposure. AB - A female patient with significant coagulopathy is presented for case discussion. This case represented a presumed Munchausen exposure to commercially available rat poison that contained one of the superwarfarin chemicals, brodifacoum. Review of the medical literature is undertaken to discuss the diagnostic approach and treatment of superwarfarin exposure. The accidental or intentional exposure to this group of rodenticide represents a significant public health problem that often is not considered by primary care physicians when confronted with coagulopathy. PMID- 19998734 TI - Health system reform. PMID- 19998735 TI - Part seven: The role of narrative in the everyday practice of medicine developing narrative competence. AB - It has been the thesis of this symposium that medicine is a narrative enterprise. We have presented our case that the work in is largely narrative. If that is true, then one of the goals of medical education should be to create methods of improving the narrative competencies in learners and practitioners of medicine. This final paper will explore the field of Narrative Medicine and briefly discuss methods currently in use in American Medical Education and conclude with the experience at the University of Oklahoma College of Medicine over the past ten years. PMID- 19998737 TI - The nursing shortage is over? PMID- 19998736 TI - Antibiotics have been revolutionary force in medicine. PMID- 19998738 TI - To mandate, or not to mandate, that is the question. PMID- 19998739 TI - What we know about influenza vaccine and Guillain-Barre syndrome. PMID- 19998740 TI - Advance directive and POLST: complements and contrasts. PMID- 19998741 TI - H1N1 and your contract: what you need to know. PMID- 19998742 TI - Top ten reasons to support National Health Care Reform. PMID- 19998743 TI - No longer "if," but "when": the coming abbreviated approval pathway for follow-on biologics. AB - Abbreviated approval of follow-on biologics involves answering complex scientific, legal, and policy questions. The Food and Drug Administration (FDA or the Agency) asserts that it lacks the statutory authority to approve follow-on versions of biologics licensed under Section 351 of the Public Health Service Act (PHSA). Despite persuasive arguments to the contrary the one hundred and tenth Congress entertained four legislative proposals to give FDA this authority, each markedly different. It is no longer a question of "if," but "when" FDA will receive authority to review and license abbreviated applications for follow-on biologics. Any legislation in the one hundred and eleventh Congress must determine: (1) if FDA should be granted authority to develop an abbreviated pathway through rulemaking or guidance; (2) if human clinical trials should be mandatory or discretionary; (3) the feasibility of interchangeability determinations in light of patient safety concerns; (4) the duration of marketing exclusivity for associated products; (5) which products are eligible for follow on approval; and (6) the degree to which uniformity is achievable between the FD&C Act and the PHSA. This paper recommends the one hundred and eleventh Congress strike a balance between patient safety, incentives for product innovation, price competition, and the need for a flexible, transparent process that capitalizes on FDA's growing expertise with follow-on biologics approvals under Section 505(b)(2) of the FD&C Act. PMID- 19998744 TI - Export of pharmaceuticals and medical devices under the federal Food, Drug & Cosmetic Act: FDA's striking change in interpretation post-Shelhigh. AB - With no communication to industry except court filings in United States v. Undetermined Quantities of Boxes of Articles of Device (Shelhigh) and a draft guidance document, the Food and Drug Administration (FDA) has articulated new policies regarding export of pharmaceutical products and medical devices. FDA's departure from its historic interpretation of the export provisions of the Federal Food, Drug, and Cosmetic Act (FDCA) significantly limits the ability of manufacturers to export misbranded drugs and medical devices that FDA deems "adulterated," contrary to the plain language and legislative intent of the FDCA. To further exacerbate the issue, FDA has begun to implement these policies without the notice-and-comment rulemaking required by the Administrative Procedures Act (APA), but rather through an enforcement proceeding brought in the United States District Court for the District of New Jersey. In a letter opinion, the District Court prevented the export of Current Good Manufacturing Practices (CGMP) --adulterated medical devices that complied with FDCA Section 801(e)(1), at least as historically interpreted by FDA. The purpose of this article is to review the history of FDA's export policies for pharmaceuticals and medical devices, particularly those aspects of the export policies that are affected by FDA's recent change in position. Three changes in FDA's interpretation of the export provisions of the FDCA will be addressed: 1) unapproved devices that a manufacturer reasonably believes are eligible for Section 510(k) clearance may no longer be exported under Section 801(e) and now must be exported under Section 802, in substantial compliance with Current CGMP; 2) adulterated devices and misbranded drugs can only be exported if the foreign purchaser's specifications cause the product to be adulterated; and 3) an article may not be exported if a like article has ever been sold or offered for sale in domestic commerce. FDA's new interpretations of FDCA Sections 801 and 802 are not valid interpretations of these statutory provisions. Although FDA always exercised its enforcement discretion under the FDCA with respect to export of an unapproved device that a manufacturer reasonably believed to be eligible for Section 510(k) clearance, FDA's recent shift in interpretation contravenes its longstanding approach without sufficient public notice. PMID- 19998745 TI - Analysis of the impact of the Uruguay Round Agreements Act on pharmaceutical patents. AB - The Uruguay Round Agreements Act (URAA) was enacted by the United States Congress in 1994. The URAA provided substantial modifications to the framework for U.S. patent law that came into effect January 1, 1953. Changes in patent regulation are especially important in the pharmaceutical sector because the patent system determines, in large part, the reward that an inventor can derive from discovery of a new drug. Most pharmaceutical patents are classified as utility patents. Pharmaceutical patents may include claims for the active ingredient per se, for the formulation of the active ingredient for use as a pharmaceutical, for therapeutic indications and uses, and for methods of manufacturing the drug. The U.S. has a First-to-Invent system to establishing the right of priority of an invention, but most countries have a First-to-File priority system. The First-to Invent system allows for public disclosure of inventions prior to patent filing. In contrast, the First-to-File system encourages early filing of patent applications. In both systems, filing an application for a patent as soon as the drug is discovered allows inventors to obtain an earlier date of invention relative to potential competitors and establish the right of priority over the invention. PMID- 19998746 TI - FDA's proposed regulations to expand access to investigational drugs for treatment use: the status quo in the guise of reform. AB - On December 14, 2006, the Food and Drug Administration (FDA) proposed two new regulations in the Federal Register amending current regulations governing expanded access to investigational drugs for treatment use and charging for investigational drugs. The proposals come at a time when FDA has found itself under new pressure to provide seriously ill patients with early access to investigational drugs outside the framework of clinical trials. In recent years, patient advocacy groups have filed citizen petitions with FDA asking the agency to provide specific criteria to patients and sponsors seeking expanded access or to create an early approval mechanism to permit easier access to investigational therapies. Further, FDA has seen proposed federal legislation intended to ensure early patient access to investigational treatments and nearly lost a lawsuit in federal court in which terminally ill patients sought a fundamental right of access to investigational therapies under the Due Process Clause of the Constitution. The proposed rules seek to assuage patient activists, physicians, drug sponsors and other critics who contend that FDA must strike an appropriate balance between allowing patient access to promising treatments while protecting against undue risk and safeguarding the clinical trials process. Although FDA heralded the announcement of the rules as a key step forward to improving patient access, the proposal does not expand access beyond measures currently available under longstanding agency practice and, in fact, creates new regulatory barriers and disincentives to industry participation in expanded access programs. This article examines the proposal in light of historical agency regulation and recent pressures to expand access. Section II describes the historical development of FDA's statutory authority to regulate drugs and the traditional new drug approval process. Section III describes the various methods through which FDA has allowed expanded access to investigational treatments since 1962. Section IV recounts various recent pressures on FDA to reform its expanded access procedures and describes the context in which FDA's recent proposal arises. Section V examines the changes proposed in both the proposed rules to expand access to investigational treatment and charging for investigational drugs. Section VI evaluates the rules and argues that the proposal will fail to expand access for patients because new restrictions on charging provide no incentive for industry participation and the proposed regulations create increased regulatory barriers to access inconsistent with FDA's statutory mandate PMID- 19998747 TI - A legal duty to disclose individual research findings to research subjects? AB - Research that utilizes human subjects is a large and growing enterprise. Tens of millions of individuals have already participated as subjects in one or more research protocols, and millions more participate each year. Government and industry combined spend billions annually to support as many as 20,000 research studies, many of which are individually large and complex enterprises in their own right.These numbers are, if anything, likely to increase even further. Besides the growth in research, two other trends are apparent. First, research related litigation is on the rise and appears likely to become even more widespread. Sparked at least in part by recent widely publicized instances of harm befalling research subjects, plaintiffs' attorneys are suing both more often and more creatively. Related to this is the second trend: public trust in research is declining and, as a result, at least some types of research are struggling to find adequate numbers of human subjects.As a result of these trends, exposure to potential liability and public perception are both increasingly important. Concomitant with all of this research is the discovery and generation of tremendous quantities of data specific to individual subjects, including--but not limited to--genetic information. Much of this data is irrelevant to subjects' interests because it lacks predictive value, has uncertain meaning, or is otherwise uninformative. Some, however, is different- some of the personal data learned during the course of research with human subjects bears directly on individuals' health. Despite the fact that much individual data has already been generated and that both the quantity and the quality of data generated seem likely to increase, there is a lack of clear guidance for researchers regarding whether and when such information should be divulged to the subjects on whom it bears.In this environment, the potential exists for litigation alleging that a researcher was negligent for failure to disclose to a subject an individual research finding of medical significance. Such litigation would raise a heretofore-unanswered question: should a researcher have a legal duty to disclose medically significant individual research findings to a subject? PMID- 19998748 TI - Genomic signatures of influenza A pandemic (H1N1) 2009 virus. AB - Adaptive mutations that have contributed to the emergence of influenza A pandemic (H1N1) 2009 virus, which can replicate and transmit among humans, remain unknown. We conducted a large-scale scanning of influenza protein sequences and identified amino acid-conserving positions that are specific to host species, called signatures. Of 47 signatures that separate avian viruses from human viruses by their nonglycoproteins, 8 were human-like in the pandemic (H1N1) 2009 virus. Close examination of their amino acid residues in the recent ancestral swine viruses of pandemic (H1N1) 2009 virus showed that 7 had already transitioned to human-like residues and only PA 356 retained an avianlike K; in pandemic (H1N1) 2009 virus, this residue changed into a human-like R. Signatures that separate swine viruses from human viruses were also present. Continuous monitoring of these signatures in nonhuman species will help with influenza surveillance and with evaluation of the likelihood of further adaptation to humans. PMID- 19998749 TI - A groundhog, a novel Bartonella sequence, and my father's death. PMID- 19998750 TI - Platelets and vascular disease. PMID- 19998751 TI - Interspecies crosses: aspects of animal protection. PMID- 19998752 TI - The effects of mental illness on labour market participation. PMID- 19998753 TI - [Patient information. Treatment of chronic hepatitis B with oral antivirals]. PMID- 19998754 TI - Paradigms and EBD. PMID- 19998755 TI - Tender muscles and masticatory myofascial pain diagnosis: how many or how much? PMID- 19998756 TI - [Terminal care and the vegetative state]. PMID- 19998757 TI - [Three dimensional echocardiography]. AB - Real time three dimensional echocardiography has arrived into clinical cardiology, allowing noninvasive and repeatable spatial visualisation and accurate quantification of cardiac movement and volumes. Evaluation of valvular disease and cardiac masses with three dimensional echocardiography is superior to traditional two dimensional imaging. Since transesophageal imaging is also available, three dimensional intraoperative echocardiography and monitoring of percutaneous procedures like ASD and PFO closures is now possible. PMID- 19998758 TI - [Encouraging experiences of interactive lectures]. AB - BACKGROUND: Traditional lectures typically represent unidirectional transfer of information from teacher to students whilst interactive lectures involve student activity. MATERIAL AND METHODS: We analyzed the experiences of students and teachers of interactive lectures by observation and questionnaires during a course organized by Helsinki Biomedical Graduate School. RESULTS: Teachers and the majority of students found interactive lectures highly motivating although we observed that only a fraction of students participated in discussions. Students were of the opinion that interactivity improved their learning. CONCLUSIONS: Supplementing lectures with interactive elements encourages students to adopt active learning techniques. PMID- 19998759 TI - [Analysis of proteinuria]. AB - Targeted urine screening surveys for risk groups are beneficial. The measurement of microalbuminuria has become an established method for the screening and monitoring of diabetic nephropathy. If strip testing has revealed albuminuria in a symptomless subject as an incidental finding, a first-voided morning urine specimen will be reassayed for albumin. If the strip test result is repeatedly positive, elucidation of the extent of proteinuria is essential. Proteinuria can be divided into glomerular, tubular and overflow proteinuria. Persistent proteinuria is an important clinical finding having significant predictive value for the health and future of a person being examined. PMID- 19998761 TI - [Update on current care guidelines. Prevention and treatment of acute kidney injury in adults]. AB - Acute kidney injury (AKI) is associated with significant morbidity and mortality. Its prevalence is increasing. Risk factors are older age, diabetes, atherosclerosis, medications, heart failure, male sex, and even mild chronic renal failure. Early detection of AKI is essential, as is the prevention of AKI related to hypovolaemia, contrast agents and nephrotoxic medications. No medication is available for developed AKI, the only therapeutic option being renal replacement therapy. Thus, prevention by adequate fluid therapy, optimisation of renal perfusion pressure and exclusion of post-renal causes of AKI are crucial. To date, the long-term outcome in AKI is unsatisfactory and the costs are high. PMID- 19998760 TI - [Use of CSF biomarkers in the diagnostics of memory diseases]. AB - Alzheimer's disease is the most common cause of dementia. Early diagnosis of diseases leading to dementia is cost-effective. However, early diagnosis of Alzheimer's disease may be difficult and should not be based on exclusion of other reasons for dementia. A decreased CSF amyloid beta-peptide-42 level together with elevated tau or phosphorylated tau levels can differentiate patients with AD from control subjects or patients with other neurologic conditions with relatively high accuracy. We evaluated the use of these biomarkers in 452 patient cohort during 2005-2007 in clinical practice. Cerebrospinal fluid biomarkers seem to be useful especially to confirm Alzheimer's disease diagnosis. PMID- 19998762 TI - [Where does obesity stand in health care--every alternative into use]. AB - Although intervention in obesity on a consultation visit, i.e. brief intervention, will only affect a portion of patients, it can reach large numbers of patients. Basic care of obesity, i.e. patient education in a group, leads to an average weight loss of 3 to 5 kg within one to two years. Basic care should be offered to patients having a condition that requires weight reduction during the treatment. In morbid obesity, surgical treatment should be promoted in Finland, although it can only be applied to a minor fraction of those requiring antiobesity treatment. Self-motivated weight control should thus be encouraged. PMID- 19998763 TI - [Effectiveness and safety of bariatric surgery]. AB - Evidence for the effectiveness of surgical treatment of morbid obesity on life span, quality of life and associated diseases proves it an applicable alternative for the morbidly obese in cases where other means of controlling obesity have remained ineffective. The operation causes significant weight reduction even within a 15-year follow-up. Surgical mortality after an endoscopic operation is usually less than 0.5%. After the operation, patients fairly often experience various symptoms of the digestive tract. Common complications include various infections and functional disturbances associated with the band. PMID- 19998764 TI - [Cost-utility of bariatric surgery in the treatment for morbid obesity in Finland]. AB - Finohta's health technology assessment report on bariatric surgery included a cost-utility analysis on three main surgical interventions used in Finland. A cost-utility analysis from the health care provider's perspective with a ten year time horizon was conducted. The parameter values were based on a representative population survey, register data, literature and expert opinions. Based on the analysis, bariatric surgery is more effective and less costly than current prevailing forms of treatment for the morbidly obese in Finland. The results were robust and consistent with previously published studies: Bariatric surgery is cost-effective in treatment of morbid obesity. PMID- 19998765 TI - [Provision of surgical treatment for the morbidly obese in Finland]. AB - The number of and demand for surgical treatments of morbid obesity are quickly increasing in Finland. The resources and action models differ regionally. The position of antiobesity surgery among antiobesity treatments should be defined more clearly than is now the case in Finland. Also a decision should be made how these operations are included in the patient's treatment guarantee. There should be a public discussion on how to centralise the operations into sufficiently large units of expertise and availability of the treatment in various parts of Finland must be guaranteed. Treatment chains should be standardized, at the same time ascertaining adequate expertise of various health care providers. PMID- 19998766 TI - [Ethical aspects of bariatric surgery]. AB - Attitude to obesity determines the attitude to antiobesity surgery: is obesity a disease among other diseases belonging to the public health care, or a characteristic resulting from living habits, remaining under the person's own responsibility? Antiobesity surgery requires supporting the patients' right to self-determination. Other treatments for obesity should be available, and this procedure necessitating permanent adjustments of living habits requires sufficient advance preparation. The patient's guarantee governing the treatment of obesity as a whole seems relevant. Assessment of the need for surgery is, however, difficult and the benefits and disadvantages differ among the various stakeholders. PMID- 19998768 TI - Th future of digital health. Six experts offer their visions. PMID- 19998767 TI - [Effects of bariatric surgery on mental and social well-being]. AB - Morbid obesity is associated with severely invalidizing symptoms and a strong stigma, which restrict the management of daily life. The handicaps of morbid obesity are so severe that patients are ready to accept even inconvenient adverse effects or high risks from the treatment. Seeking antiobesity surgery is mainly done for health reasons, but is perceived to have a broad effect on functional capacity, self-image as well as on mental and social well-being. By the help of anti-obesity surgery, many patients not only gain control over eating, but also over other fields of life. PMID- 19998769 TI - Are HIEs the answer? Alternatives to regional health information exchanges are emerging. PMID- 19998770 TI - Measuring what works. A community hospital dives into data to come up with cardiac best practices and guide treatment decisions. PMID- 19998771 TI - A team effort. The CIO and CMIO at Orlando Health make sure doctors are involved in I.T. PMID- 19998772 TI - Second time's the charm. Rural hospital involves physicians in its latest effort to roll out EHRs at clinics. PMID- 19998773 TI - Card player. Interview by Elizabeth Gardner. PMID- 19998774 TI - [Culture of safety--the magic bullet for patient safety?]. PMID- 19998775 TI - [Safety culture: definition, models and design]. AB - Safety culture is a multi-dimensional phenomenon. Safety culture of a healthcare organization is high if it has a common stock in knowledge, values and symbols in regard to patients' safety. The article intends to define safety culture in the first step and, in the second step, demonstrate the effects of safety culture. We present the model of safety behaviour and show how safety culture can affect behaviour and produce safe behaviour. In the third step we will look at the causes of safety culture and present the safety-culture-model. The main hypothesis of this model is that the safety culture of a healthcare organization strongly depends on its communication culture and its social capital. Finally, we will investigate how the safety culture of a healthcare organization can be improved. Based on the safety culture model six measures to improve safety culture will be presented. PMID- 19998776 TI - [Safety culture in the context of work intensification--development in Germany over the last 10 years]. AB - This article tries to review the development of patient safety culture in the German healthcare system over the last decade. Since the use of standardized questionnaires and other instruments to measure safety culture in Germany has only just begun there are no representative and longitudinal data. Therefore a set of indicators and clues is chosen to characterise the safety culture development on the micro-, meso- and macro-level of the healthcare system in four areas. Is patient safety an issue of the healthcare debates and especially of research? Have dedicated structures and processes been implemented to support clinical risk management? What are the objective outcomes of healthcare and treatment in regard to patient safety? In summary, there are a lot of signs that patient safety issues in Germany are gaining more and more importance on all levels of the healthcare system. To date there have been single evidence-based studies only indicating a causal or close temporal relationship between patient safety outcomes and the increasing efforts of hospitals, outpatient and long-term care facilities. PMID- 19998777 TI - [Organisational responsibility versus individual responsibility: safety culture? About the relationship between patient safety and medical malpractice law]. AB - The contribution is concerned with the correlations between risk information, patient safety, responsibility and liability, in particular in terms of liability law. These correlations have an impact on safety culture in healthcare, which can be evaluated positively if--in addition to good quality of medical care--as many sources of error as possible can be identified, analysed, and minimised or eliminated by corresponding measures (safety or risk management). Liability influences the conduct of individuals and enterprises; safety is (probably) also a function of liability; this should also apply to safety culture. The standard of safety culture does not only depend on individual liability for damages, but first of all on strict enterprise liability (system responsibility) and its preventive effects. Patient safety through quality and risk management is therefore also an organisational programme of considerable relevance in terms of liability law. PMID- 19998778 TI - [Patient safety and a culture of responsibility in ambulatory care: strategies for improving practice]. AB - The development of a medical safety culture is spreading beyond the hospital into the ambulatory setting. Patient safety defined as "absence of unwanted events" (primum non nocere) can serve as a starting point for the advancement of our ambulatory medical care system. Error analyses conducted in GP and specialist practices will identify gaps and traps in the system and provide ideas for the development and implementation of new safety strategies in ambulatory patient care. In the light of the structures and processes of GP medical care aspects of patient safety will be correlated to the outcome quality and examples will be discussed. Possible strategies for the improvement of patient safety in GP practice will be presented from the perspective of both patient- and practice individuality. PMID- 19998779 TI - [Is patient safety culture measurable and if so, how is it done?]. AB - Safety culture is one aspect of organisational culture. It is the pattern of shared values, beliefs and attitudes concerning safety present in an organisation. In this article, Edgar Schein's three-layer model is used to demonstrate to what extent a safety culture can be directly and indirectly measured. Several methods for the assessment of safety culture are presented: questionnaires evaluating the safety climate, methods enabling teams to perform self-assessment, interviews, audits, analyses of safety reports and observation. The aim of the assessment is crucial to the choice of the appropriate mode of measurement. In healthcare, the currently most widely used instrument is questionnaires since it is comparatively simple to use and inexpensive and yields rapid results. However, the employment of additional measures (interviews, audits, analyses of safety reports) is more appropriate if the most important aspects of safety culture in a given organisation need to be covered. PMID- 19998780 TI - [Developing a questionnaire to assess the safety climate in general practices (FraSiK): transcultural adaptation--a method report]. AB - BACKGROUND: Safety climate comprises shared perceptions of safety issues and is a visible feature of a safety culture. Usually self-report questionnaires are used to assess a safety climate. The aim of this project was to develop the first German questionnaire for the assessment of the patient safety climate in general practices. METHODS: The development of the instrument was based on a US questionnaire. The ambulatory care setting differs considerably between the two countries in terms of organisational and economic issues as well as the number and variety of healthcare professionals. The process of adaptation therefore required the following steps: 1) translation of the Safety Attitudes Questionnaire, ambulatory version (SAQ-A), 2) interviews with experts on quality improvement in order to evaluate the comprehensiveness and relevance of the SAQ-A in the new context, 3) revision of items, 4) pretesting of the preliminary version on a sample of healthcare professionals from general practices and 5) a final revision. RESULTS: The "Fragebogen zum Sicherheitsklima in Hausarztpraxen" (FraSiK) is a questionnaire consisting of 68 items, 36 of which were adapted from the original. Altogether, 32 items, relating to the involvement of patients, communication, education and training, error management and dealing with regulations were added. CONCLUSION: Substantial changes were made to the original questionnaire and items added in order to obtain an appropriate instrument for the assessment of the content and relevance of patient safety culture in the socio-cultural context of German general practices. PMID- 19998781 TI - [Quality management and safety culture in medicine: context and concepts]. AB - The publication of the IOM report "To err is human: building a safer health system" in 1999 put spotlight on the primacy of the principle of primum non nocere and made patient safety a central topic of quality management. A key conclusion of the report was the need for a well-developed safety culture. While concepts of quality management have evolved along the lines of ISO and Total Quality Management over the last decades patient safety still has not got the same amount of attention (PubMed). Evidence from other safety-critical areas but also from the field of medicine itself suggests that an efficient culture of safety is a conditio sine qua non for the sustainable improvement of patient safety. Considering these arguments the present paper aims at emphasizing the importance of an efficient culture of safety for patient safety and quality management in healthcare. In addition, key instruments of safety culture as well as their limitations will be presented. PMID- 19998782 TI - [Comparison of quality by means of funnel plots--a plea for a uniform methodology]. AB - Vertical bar charts depicting unit event rates sorted in ascending order enjoy widespread usage in external quality assurance. Unfortunately they suggest a spurious ranking resulting from instability in the percentile distribution chiefly caused by varying denominators. The popular remedy of simply excluding units below a minimum threshold would solve the problem only partially since units with few operations per annum may evade evaluation altogether merely by the grace of their size. Compared with alternative solutions reviewed in this article Spiegelhalter's funnel plots exhibit clear advantages over statistical control charts or Bayesian modelling. A major drawback of control charts at present is that data are still widely transferred on a yearly rather than a quarterly or even monthly basis. The chief disadvantage of Bayesian modelling lies in the difficulty of obtaining consensus on the required prior distributions. Funnel plots on the other hand provide a flexible and sample size dependent uniform approach while at the same time offering an intuitive interpretation of volume effects. The addition of control and warning limits allows for formal assessment of deviations from target values. PMID- 19998783 TI - [League of Nations: medical guidelines in Austria]. PMID- 19998784 TI - [Occupational ear diseases]. PMID- 19998785 TI - [Germany-induced changes in the consequences of organizations, professionals, patient and quality--practical care research with amazing results]. PMID- 19998786 TI - [Letter on the publication "Culture of safety in concentrated work conditions"- development in Germany in the last 10 years]. PMID- 19998787 TI - Forensic aspects of reckless prescribing. PMID- 19998788 TI - Rape investigation--a policing perspective. PMID- 19998789 TI - Medico-legal issues: a view from the largest acute trust in England. PMID- 19998790 TI - Lethal witness. PMID- 19998791 TI - A note on terminally-ill teenagers. PMID- 19998792 TI - [Interest of immunohistochemic markers (Ki67, HMB45, p53) in risk analysis of congenital naevi of little and middle size]. AB - The risk to develop melanoma from small or medium size congenital naevus remain controversial. The main goal of the present study was to determine the interest of three immunohistochemical markers (Ki67, HMB45 and p53) in predicting malignant transformation of these congenital naevi and to see if a specific immunohistochemical profile of such transformed naevi can be identified. The markers (Ki67, HMB45 and p53) have been used retrospectively on sections of small or medium size congenital naevi (group NC, n = 15), of melanoma developed on small or medium size congenital naevi (group MNC, n = 15) and of melanoma developed on acquired naevi (group MNA, n = 15). The labelled cells have been counted in different cutaneous layers: junction, superficial dermal layer and deep dermal layer. No reactivity was observed for the three markers in group NC. The percentage of labelled cells was significantly different for the three markers between the group NC and the groups MNC and MNA. There was no difference between the groups MNC and MNA. In the groups MNC and MNA, a gradient in the percentage of labelled cells was observed between superficial and deep layers. These three markers do not differentiate melanoma developed from congenital naevi of small or medium size and melanoma developed from acquired naevi. Moreover, the results suggest that these three markers are useless in predicting the risk of malignant transformation of small or medium size congenital naevi. PMID- 19998793 TI - [Parkinson syndromes in geriatrics]. AB - Geriatric teams are more and more in front of patients presenting Parkinson syndromes. The frailty of the geriatric patient could be responsible of the non recognition of the diagnosis of the Parkinson and related diseases. A diagnostic approach is proposed in order to better define a care plan and to target the priorities. In collaboration with the general practitioner, the neurologist and the caregivers, an appropriate diagnosis allows to optimize and/or to adapt the treatment, to prevent the functional decline, to decrease the caregiver's burden, and eventually to improve the end of life. PMID- 19998794 TI - [Oncogeriatrics: an attempt to operational definition]. AB - Comprehensive geriatric assessment (CGA) represents a multidisciplinary comprehensive evaluation of an older individual's functional status, comorbid medical conditions, cognition, psychological state, social support, nutritional status, and a review of the patient's medications. Initially, the use of a CGA in the care of older patients with cancer was based on an extrapolation of its ability to predict morbidity and mortality in the general geriatric population. More recently, however, accumulating data show the benefits of using a CGA particularly in patients with cancer to predict morbidity and mortality. Prospective trials evaluating the utility of a CGA to guide interventions to improve the quality of cancer care in older adults are justified. PMID- 19998795 TI - [Glioblastoma treatment in 2010]. AB - The treatment of glioblastomas requires a multidisciplinary approach because despite the progresses in surgical and iconographic managements associated with research knowledge this disease presently remains incurable and progresses during the 6 months after its diagnose. Current recommendations are that patients with glioblastoma should undergo maximum surgical resection followed by concurrent radiation and chemotherapy with the alkylating drug temozolomide, followed subsequently by additional adjuvant temozolomide for a period of up to 6 months. Temozolomide mechanism of action is complex and we have recently evidenced a temozolomide-associated anti-angiogenic activity in vitro and in vivo on preclinical human glioblastoma models. We describe in the current review the temozolomide-associated antiangiogenic activity. We also describe here the major signaling pathways that can be constitutively activated in migrating glioma cells, and which render these cells resistant to proapoptotic insults such as conventional chemotherapies. In light of this resistance, we therefore describe the targeted therapies and local drug delivery systems which could be used to complement conventional treatments. We have reviewed more than 400 ongoing clinical trials with respect to these new targeted therapy approaches alone or in combination for glioblastoma therapy and we also emphasize the importance of vaccinotherapy. We conclude our review with a therapeutic model that could be used in the light of the present knowledge. PMID- 19998796 TI - [Meningitis and renal cancer]. AB - A 51 year-old man with a renal cancer (pT3N0M0) consulted at the emergency department for persistent headache initiated a week before and recent vomiting. Physical examination and blood analyses were not remarkable. Cerebral magnetic resonance imaging demonstrated radiological signs of meningitis. The differential diagnosis of meningitis is discussed. PMID- 19998798 TI - [Recurrent maxillo-mandibular odontogenic keratocysts: prevention and treatment]. AB - Keratocyst is an odontogenic cyst with an aggressive clinical behavior and a high recurrence rate. Our work describes a 62 year-old patient with Gorlin's syndrome presenting advanced recurrent maxillo-mandibular keratocysts. He already had multiple resections of these lesions. Early diagnosis and treatment of keratocysts is crucial to reduce the extension of the cysts and minimize their malignant transformation especially when they are part of a Gorlin's syndrome. PMID- 19998797 TI - [Extrafollicular adenomatoid odontogenic tumour: report of two cases]. AB - The purpose of this study was to analysis the clinicopathological features of adenomatoid odontogenic tumour extrafollicular and to discuss the diagnosis pitfull. We present two cases diagnosed and followed at the C.H.U. Habib Bourguiba, Sfax. It is about a 15 year-old boy and a 46 year-old woman who presented gingival swelling. The panoramic radiographs showed in both cases, a radiolucent area affecting teeth 23, 24 and 35 respectively. A computed tomography scan, realized at the second cas, demonstrated an expansile lesion with a sclerotic rim. An exploratory surgical approach was chosen and the final diagnosis was microscopically confirmed to be an extrafollicular variant of adenomatoid odontogenic tumour. The patients were healthy and have not shown any signs of recurrence at follow-up. The two cases described illustrate clinical and radiographic features of the extrafollicular variant of adenomatoid odontogenic tumour. Careful diagnostic procedure and adequate interpretation of radiographic findings may result in a correct diagnosis with otherwise may result in unnecessary endodontic treatment. PMID- 19998799 TI - [Thrombocytopenia and acute kidney injury]. AB - A 31-year old man is addressed to the emergency department for fever, abdominal pain and vomiting. Laboratory tests reveal an inflammatory syndrome, thrombocytopenia, acute kidney injury associated with major proteinuria (more than 3 g/24 h). Evaluation will conclude to an acute interstitial nephritis and the serology is positive for Hantavirus. This clinical case illustrates differential diagnosis of acute renal failure and reminds the theory of hemorrhagic fever with renal syndrome : usual clinical presentation is "flu"-like symptoms, thrombocytopenia and acute renal failure. Hantavirus is endemic in some area in Europe and in Belgium. This clinical presentation should suggest the diagnosis if the patient has stayed in an endemic area. PMID- 19998800 TI - [A ventricle of all dangers: a clinical observation and evolution of a case of arrythmogenic right ventricular dysplasia]. AB - Arrythmogenic right ventricular dysplasia (ARVD) is rare disease but it is a major cause of sudden death in young people and in athletes. ARVD is a cardiomyopathy characterized by structural and functional abnormalities of the right ventricle precipitating its electrical instability. This electrical instability is responsible for ventricular arrhythmias. Sudden death, by ventricular fibrillation, may be the first symptom of the disease. The diagnosis is based upon specific ventricular pathology at the histological level as well as typical electrocardiographic and diagnostic imaging features. A better understanding of the disease has increased along with the new data on genetics. Its management remains a challenge because of the wide spectrum of clinical presentation as well as its natural history. We present a case of ARVD and its evolution in order to show the difficulties of the management of this particular entity. PMID- 19998801 TI - [Medicinal leeches]. AB - Leeches are hermaphroditic and hematophagous annelids. One important medical species, Hirudo medicinalis, comes from hirudiniculture of fresh water pools. Thanks to their three mandibles with some 300 teeth on their anterior muscular sucker, they easily grab to tissues and by secreting their saliva containing numerous powerful enzymes, such as hyaluronidase, collagenase and inhibitors of platelet aggregation and coagulation, like hirudin, allow blood sucking. Once they are full of blood (up to 15 g of blood), they detach themselves from their prey. Used ever since the 18th Egyptian Dynasty, leeches became famous during the first part of the XIXth century, thanks to a French physician, Francois Joseph Victor Broussais, known to his adversaries as the "vampire of medicine" for treating various conditions such as phlebotomy, laryngitis, ocular problems, obesity, mental disorders, etc. Overfishing, therapeutic failures and most particularly, the emergence of hygiene, brought the decline of living leeches. In 1884, an extract of leeches was obtained--hirudin and henceforth used. Nowadays, leeches are still used in microsurgery to enhance the venous circulation in finger reimplantation or skin flap transposition. Hirudin is synthesized through recombinant DNA technology and molecules such as lepirudin and desirudin are available on the market as anticoagulant. PMID- 19998802 TI - [Ibero-American cardiovascular journals. Proposals for a necessary collaboration]. PMID- 19998803 TI - Association between red blood cell distribution width and outcomes at six months in patients with acute coronary syndromes. AB - BACKGROUND: Higher values of red ceildistribution width (RDW) may be associated with adverse outcomes in patients with heart failure and in those with stable coronary artery disease. We assessed the hypothesis that higher RDW values are associated with adverse cardiovascular outcomes in patients with acute coronary syndromes (ACS). METHODS: We studied 1796 patients with ACS admitted to a coronary care unit. We analyzed clinical and laboratory characteristics, management, and outcomes of patients according to tertiles of baseline RDW. The primary outcome was death or myocardial infarction (MI) during six-month follow up. RESULTS: Patients with higher RDW values tended to be older, were more likely to be female and have a history of MI, and more often had renal dysfunction, anemia, and Killip class >I on admission (p < 0.05). Higher RDW values were associated with increased 6-month mortality (tertile 1: 8.2%; tertile 2: 10.9%; tertile 3: 15.5%; p = 0.001 for trend) and increased 6-month death/MI rates (tertile 1, 13.0%; tertile 2, 17.2%; tertile 3, 22.9%; p < 0.0001 for trend). An association between higher RDW and increased 6-month death/MI rates was found in patients with non-ST-elevation ACS (10.5% vs. 15.3% vs. 22.7%; p < 0.001 for trend), with a tendency in patients admitted with ST-elevation MI (15.1% vs. 19.1% vs. 23.1%; p = 0.053 for trend). After adjustment for baseline characteristics and treatment, higher RDW values remained independently associated with the study's primary composite outcome but not with all-cause death. Using the first tertile of RDW as reference, the adjusted odds ratio (OR) for 6-month death/MI among patients in the highest RDW tertile was 1.43 (95% confidence interval [CI], 1.00-2.05; p = 0.049). Using RDW as a continuous variable, the adjusted OR for 6-month death/MI was 1.16 (95% CI, 1.03-1.30; p = 0.017) per 1% increase in RDW. CONCLUSIONS: RDW is an easily determined predictor of outcome after ACS. We found a graded independent association between higher RDW values and adverse outcomes in patients with ACS. PMID- 19998804 TI - Biomarkers in risk stratification of acute coronary syndromes: new insights into hematologic parameters. PMID- 19998805 TI - Cardiac tumors: the 17-year experience of pediatric cardiology department. AB - INTRODUCTION: Primary cardiac tumork may be benign or malignant and may arise anywhere in the endocardium, myocardium or pericardium. They are rare in pediatric age-groups, with a prevalence in autopsy findings of 0.0017 to 0.28%; most are benign, malignant tumors accounting for only 10% of the total. OBJECTIVE AND METHODS: The purpose of this paper is to report the 17-year (1989-2006) experience of primary cardiac tumors in a single center. The clinical data and histopathological diagnoses were collected from a retrospective chart review. RESULTS: We identified 12 patients, with a mean age at diagnosis of 23 months, in two cases diagnosed in the fetal period. The frequent cause of referral was tuberous sclerosis, followed by a cardiac murmur. The diagnosis of primary cardiac tumor was based on echocardiography and/or magnetic resonance imaging. The patients all underwent 24-hour electrocardiographic Holter monitoring, and the most common finding was nonspecific repolarization abnormalities. The most frequent cardiac tumor was rhabdomyoma (67%), usually associated with tuberous sclerosis, followed by fibroma (17%) and fibroelastoma (8%) Although biopsy is the diagnostic gold standard, it was only performed in two patients. Surgical removal of the tumor was performed in a single patient (with fibroelastoma) due to the perceived risk of pulmonary embolization. DISCUSSION AND CONCLUSIONS: In our population, the most frequent tumor was rhabdomyoma associated with tuberous sclerosis. Most patients were asymptomatic, and the diagnosis was made in routine observations on patients with tuberous sclerosis or with a cardiac murmur. A histopathological diagnosis is only warranted in the rarest forms of cardiac tumors. PMID- 19998806 TI - Which echocardiographic definition should be used to define response to cardiac resynchronization therapy? AB - INTRODUCTION: The definition of response to cardiac resynchronization therapy (CRT) remains controversial, with different criteria being used to define a positive response. The PROSPECT trial recently demonstrated that echocardiography is not sufficiently accurate to identify responders to CRT. However, it is possible that the definition used in this study was not the most appropriate. OBJECTIVE: To compare different echocardiographic definitions of response to CRT with peak oxygen consumption (VO2), in an attempt to identify the best echocardiographic definition. METHODS: Thirty consecutive patients who underwent echocardiography and cardiopulmonary exercise testing (CPET) before and 6 months after CRT were studied. An improvement of > or =1 NYHA class defined clinical responders; a > or =15% decrease in left ventricular end-systolic volume (LVESV) defined remodeling responders; a > or =25% improvement in left ventricular ejection fraction (LVEF) identified responders according to LVEF; a >25% improvement in left ventricular dP/dt defined responders according to dP/dt; and a ?10% improvement in peak VO2 defined CPET responders. RESULTS: There were 47% responders according to the reverse remodeling definition, 60% according to LVEF and 67% according to dP/dt; 77% were clinical responders and 40% CPET responders. The only baseline characteristic that differed between CPET responders and non responders was the sphericity index (57 +/- 12% vs. 72 +/- 16%, p = 0.019), which showed an inverse correlation with CPET response (r = -0.455, p = 0.011). LVEF response showed the best agreement with CPET response (83% positive and 56% negative concordance). Clinical and echocardiographic responses were often discordant: 48% of clinical responders were non-responders according to reverse remodeling, 35% according to LVEF and 39% according to dP/dt. However, of clinical responders who did not respond on echocardiographic criteria, a positive NYHA response paralleled the CPET definition in 35% of cases. CONCLUSION: The best agreement between echocardiographic definitions of response and CPET was achieved with LVEF. In 35% of cases of discrepancy between clinical and echocardiographic responses, the clinical response paralleled CPET, which implies a benefit of CRT undetected by echocardiography and not a placebo effect. PMID- 19998808 TI - The role of antidiuretic hormone in the pathophysiology and treatment of heart failure. AB - Heart failure (HF) is still one of the most important causes of morbidity and mortality worldwide. Neurohormonal changes appear to play an important role in the development and continuation of HF. Among the mediators responsible for these changes, antidiuretic hormone (ADH) is probably the least known. However, elevated concentrations of ADH are frequently found in this syndrome and have prognostic value in addition to known biomarkers. Recent experimental studies and clinical trials have aroused interest in the possible benefits of ADH receptor antagonists. This article reviews the pathophysiological mechanisms of ADH in HF and the latest advances in ADH antagonism in the therapeutic management of HF. PMID- 19998807 TI - Cardiac resynchronization therapy--clinical and echocardiographic characteristics of responders and exceptional responders. AB - Cardiac resynchronization therapy (CRT) is a novel treatment for systolic heart failure and in successful cases reverse remodeling occurs with reduction of left ventricular (LV) dimensions and volumes. The term "responders" applies to all patients who improve in functional class and quality of life; however, some responders show exceptional improvement, including normalization of clinical and echocardiographic parameters (exceptional or super-responders). OBJECTIVE: The purpose of our study was to analyze responders (R) and super-responders (SR). METHODS: 36 patients were referred for CRT due to depressed left ventricular function (mean ejection fraction 26 +/- 6.9%) and QRS duration (QRSd) of 175 +/- 29.9 ms. All patients underwent complete 2D, Doppler, and tissue Doppler (TDI) echocardiography before and after CRT (performed 3-6 months after CRT implantation) and measurement of pro-BNP. Twelve were non-responders (nR--Group I). Of the 24 patients showing improvement in NYHA functional class (responders), 11 were classified as super-responders with ejection fraction > or =40% (Group II -13 patients [R] and Group III--11 patients [SR]). RESULTS: There were no differences between the three groups in QRSd or the following baseline echocardiographic parameters: LV dimensions and volumes, ejection fraction, TDI velocities and dyssynchrony parameters. The exception was left atrial (LA) dimensions (greater in group I) and mitral inflow, with a predominant restrictive pattern in Group I. Baseline maximum VO2 was lower in Group I compared to Group III (13 vs. 21 mm/kg/min, p < 0.0001). We compared the three groups during a mean follow-up of 15 +/- 8 months. Group II showed slight improvement of left ventricular function without statistical significance, only pro-BNP showing a significant reduction (p = 0.04). Comparing Groups I and III, there were differences in the super-responder group for pro-BNP and echocardiographic LV dimensions, volumes and ejection fraction, LA dimensions, pulmonary artery pressure and TDI velocities, with normalization of septal systolic velocity: Group III--5.13 cm/s vs. Group I--2.95 cm/s, p = 0.001. CONCLUSION: The degree of improvement in CRT patients is difficult to predict by baseline echocardiography and QRSd, only a restrictive mitral inflow pattern appearing to be a determining factor. Remodeling in super-responders is probably due to a structural process, with a trend towards normalization of ejection fraction and other systolic parameters. PMID- 19998809 TI - Primary angioplasty of the left main coronary artery with culotte skenting in non ST elevation acute coronary syndrome. AB - Treatment of non-ST elevation acute coronary syndrome has long been the subject of debate and study, particularly with regard to the timing of coronary angiography and angioplasty. According to the latest European guidelines, an emergent strategy should be adopted in certain patients (estimated at 5%) with hemodynamic instability, malignant arrhythmias or angina refractory to optimal medical therapy. The authors report a case illustrating this situation in which the lesion severity and anatomy required a complex procedure of dilation of the left main coronary artery ending with a double-stent technique. PMID- 19998810 TI - Pharmacogenetics and anticoagulant therapy: two cases of genetically determined response to warfarin. AB - Inter- and intra-individual variability of response to warfarin means that its anticoagulant effect must be monitored, given the risk of thromboembolic complications and bleeding. This variability is influenced by gender, age, body mass index, smoking, diet, comorbid conditions, drug interactions and genetic factors. Pharmacogenetics refers to the study of genetic background to predict drug response, effectiveness and risk of adverse effects in a given patient. The authors illustrate its relevance in two case reports. A 40-year-old woman admitted for massive pulmonary thromboembolism underwent anticoagulant and fibrinolytic therapy, following which warfarin was needed in unusually high doses to achieve effective anticoagulation. The genetic variants c.430CC and c.1075AA of the CYP2C9 gene were identified, predisposing to rapid warfarin metabolism, as well as the c.-1639GG variant of the VKORC1 gene, associated with low sensitivity to the drug. Together, these variants give high resistance to warfarin. In the second case, a 76-year-old man with permanent atrial fibrillation developed excessive prolongation of prothrombin time after being treated with 5 mg/day warfarin for 5 days. The genetic variants c.430CC and c.1075AC of the CYP2C9 gene and 1639AA of the VKORC1 gene were identified. Together, these polymorphisms confer high sensitivity to warfarin, necessitating smaller doses to maintain therapeutic anticoagulation levels. The authors review the relevance of the study of genetic polymorphisms related to anticoagulant therapy and discuss its potential usefulness in clinical practice. PMID- 19998811 TI - The role of cardiovascular magnetic resonance in the evaluation of chest pain etiology: a case report. AB - In the evaluation of patients with chest pain it is crucial to differentiate between cardiac and non-cardiac etiology. In turn, acute chest pain of cardiac origin includes many clinical entities. The combination of elevated cardiac serum markers, particularly troponin, with typical clinical presentation and electrocardiographic abnormalities most frequently establishes a diagnosis of acute coronary syndrome. However, when coronary angiography reveals only non significant lesions, this may present a diagnostic dilemma. The authors present the case of a patient admitted with chest pain, initially interpreted as an acute coronary syndrome, based on laboratory parameters and electrocardiographic abnormalities. Coronary angiography showed normal coronary arteries. This result, combined with the findings of subsequent cardiovascular magnetic resonance (myocardial edema and subepicardial delayed enhancement), established a diagnosis of myocarditis. The case illustrates the application of this noninvasive imaging technique in the diagnostic work-up of patients with acute chest pain. PMID- 19998812 TI - Percutaneous closure of a large coronary fistula. PMID- 19998814 TI - [The registration bog--a consequence of old-fashioned medical record-keeping]. PMID- 19998813 TI - [Guidance in ethical balancing]. PMID- 19998815 TI - [The proposal for dementia care guidelines. The National Board of Health and Welfare welcomes the scrutiny]. PMID- 19998816 TI - [More and more live longer than 65 years. Fewer smokers and more people with healthy hearts probable explanations]. PMID- 19998817 TI - [Not only RSV can cause bronchiolitis in small children. Human metapneumovirus maybe the the second most common cause]. PMID- 19998818 TI - [Pain in legs not the same as varicose veins]. PMID- 19998820 TI - [Previously rarities, now common diseases]. PMID- 19998819 TI - [The effect of warfarin was potentiated by an antitussive. Cocillana-Etyfin increased the PK(INR) value]. PMID- 19998822 TI - [Strong increase of IBD--still not clear why]. PMID- 19998821 TI - [Polygenetic disturbance in the front line mucosa defense]. PMID- 19998823 TI - [The diagnosis of IBD must have a wide foundation]. PMID- 19998824 TI - [Current drug therapy in Crohn disease--national guidelines]. PMID- 19998825 TI - [Management of severe attack of ulcerative colitis]. PMID- 19998826 TI - [Colitis cancer--myth or reality?]. PMID- 19998827 TI - [Surgery--indispensable complement ot drug therapy]. PMID- 19998828 TI - [Reduced quality of life during attacks lays the foundation for alternative therapy]. PMID- 19998829 TI - [Swibreg--a new version of national IBD registry]. PMID- 19998830 TI - [Investigation-induced anxiety worse than premature death in breast cancer?]. PMID- 19998831 TI - [New drug in atrial fibrillation--how does it function in regular health care?]. PMID- 19998832 TI - [Still strong reason for enthusiasm for a new stroke-preventing drug]. PMID- 19998833 TI - [Relay doctors are just the top of an iceberg]. PMID- 19998834 TI - The way ahead for Wodonga. AB - In this adapted version of an article from The Australian Hospital Engineer, Kim Bruton, MIHEA, the magazine's editor, and chief engineer at the Wodonga Hospital campus of the newly-formed Albury Wodonga Health Service in Victoria, Australia, describes a project to both analyse the impact of regular maintenance on plant performance, and significantly reduce energy consumption across the hospital estate in the future. PMID- 19998836 TI - A model approach advocated. AB - Melvyn Langford, a former NHS estates and facilities manager with a number of Trusts and now an independent risk management consultant, describes the development and subsequent application of a model designed to help NHS Trusts, including their estates and facilities functions, to identify, monitor and manage key risks which, if not addressed, could impact on business continuity and patient care. PMID- 19998835 TI - Building cooperative care communities. AB - Four of the individuals behind an unusual Dutch project to bring together, under one overall "umbrella", a range of integrated healthcare, social care, educational and leisure facilities within a multicultural, multi-activity centre located in one of Amsterdam's most socially diverse "problem" districts, describe how the project evolved, and its ambitious short- and long-term goals. PMID- 19998837 TI - FM 'in fashion' now and for the future. AB - With estates and facilities "very much in fashion" given that patients now increasingly put cleanliness and the "general environment" at the top of their agenda, estates personnel now have an opportunity to be "right at the heart of health", said Health Facilities Scotland (HFS) director Paul Kingsmore, keynote speaker at the recent 2009 Healthcare Facilities Consortium conference in Oxford. However, achieving this goal would, he said, require not only developing firm goals and a strong, skilled workforce for today, but equally for the next decade and beyond. Jonathan Baillie reports. PMID- 19998838 TI - Atrium and artwork appeal to the heart. AB - The new Bristol Heart Institute (BHI), part of University Hospitals Bristol NHS Foundation Trust (UH Bristol), and one of a number of specialist regional cardiology and cardiac surgery centres built in England over the past decade in line with Department of Health regional cardiac provision strategy, opened to its first patients in May. Lead architect at CODA Architects Craig Bennett told Jonathan Baillie the "excellent spirit of collaboration" throughout the facility's design and construction was a major contributor to its completion on time and on budget. PMID- 19998839 TI - Fine tuning brings noteworthy success. AB - Over the past year the estates and facilities team at London's UCLH NHS Foundation Trust and Interserve Consulting have jointly analysed energy consumption at University College Hospital to identify saving opportunities and potential carbon footprint reduction measures. The project, under which measures proposed could see electrical energy demand cut by 25%, is just part of a wider drive to reduce carbon emissions and cut energy bills at one of London's largest hospitals. Jonathan Baillie reports. PMID- 19998840 TI - African adventure cues new direction for Keith. AB - A specialist in X-ray equipment, who spent over 30 years working for X-ray system manufacturers, IHEEM North Western branch member Keith Feay subsequently established his own consultancy. Two years ago, however, his life took an interesting new direction when a charity supplying medical equipment to hospitals in Malawi told him it required precisely his skills. Here he recounts some of his many interesting experiences repairing and maintaining equipment in Africa. PMID- 19998841 TI - [Consensus document on endomyocardial biopsy of the Associazione per la Patologia Cardiovascolare Italiana]. AB - The Italian Scientific Associations of Cardiologists and Cardiovascular Pathologists have produced this consensus document on the diagnostic role of endomyocardial biopsy (EMB) in terms of techniques, analysis and reporting. The document is intended for clinical cardiologists, hemodynamic experts, electrophysiologists, surgical pathologists, and cardiac surgeons. It has three main aims: a) to collocate EMB in the context of currently available tools for diagnosis of heart diseases; b) to provide recommendations for rational implementation; c) to outline key characteristics (standards) for Italian cardiology and surgical pathology centers that perform and analyze EMB. A general lack of prospective, controlled studies addressing EMB prohibited the use of traditional evidence-based recommendations that rely on classes of available evidence. Thus, it was agreed that three key points should be taken into account: a) the specific pathology to be diagnosed (or excluded); b) the existence of any alternative, non-invasive diagnostic techniques; c) the overall consequences of reaching a definite diagnosis on patients' clinical management. Accordingly, we propose recommendations for EMB based on the following levels of diagnostic value: level 1: no alternative method exists to reach a definite diagnosis that can have obvious consequences for clinical management; level 2a: no alternative method exists to reach a definite diagnosis; however, the implications for clinical management are uncertain; level 2b: no alternative method exists to reach a definite diagnosis; however, the diagnosis would not influence clinical management; level 3: an alternative method exists to reach a definite diagnosis. The second part of the document proposes current protocols for the preparation, analysis and reporting of EMB in the context of each main pathologic entity. Particular attention is given to tissue characterization and implementation of molecular tests. PMID- 19998842 TI - Physical activity surveillance: providing public health data for decision makers. PMID- 19998843 TI - Staying on task: challenges of global physical activity surveillance. PMID- 19998844 TI - Progress and pitfalls in the use of the International Physical Activity Questionnaire (IPAQ) for adult physical activity surveillance. PMID- 19998845 TI - Prevalence of self-reported aerobic physical activity among U.S. States and territories--Behavioral Risk Factor Surveillance System, 2007. AB - BACKGROUND: Accurate surveillance data on physical activity prevalence is important for U.S. states and territories as they develop programs and interventions to increase physical activity participation. METHODS: Using 2007 data from the Behavioral Risk Factor Surveillance System, we estimated the percentage of U.S. adults in each U.S. state and territory who met minimum aerobic activity criteria using the 2008 Physical Activity Guidelines for Americans (2008 Guidelines) and the Healthy People 2010 criteria for physical activity. SUDAAN was used to calculate prevalence estimates and 95% confidence intervals. RESULTS: The estimated prevalence of recommended aerobic activity in U.S. states and territories ranged from 44.5% to 73.3% according to 2008 Guidelines and from 30.8% to 60.0% according to Healthy People 2010 criteria. Absolute percent differences in prevalence among U.S. states and territories ranged from 11.7% to 19.1%, and relative percent differences ranged from 20.8% to 44.6%. CONCLUSIONS: In all U.S. states and territories, a larger proportion of U.S. adults met minimum aerobic activity criteria in the 2008 Guidelines than met corresponding criteria in Healthy People 2010. This difference, however, does not reflect an actual change in the amount of aerobic activity, but a change to the criteria for meeting 2008 Guidelines. PMID- 19998846 TI - Differences in physical activity prevalence and trends from 3 U.S. surveillance systems: NHIS, NHANES, and BRFSS. AB - BACKGROUND: Three U.S. surveillance systems-National Health Interview Survey (NHIS), National Health and Nutrition Examination Survey (NHANES), and Behavioral Risk Factor Surveillance System (BRFSS)--estimate physical activity prevalence. METHODS: Survey differences were examined qualitatively. Prevalence estimates by sex, age, and race/ethnicity were assessed for comparable survey periods. Trends were examined from NHIS 1998 to 2007, NHANES 1999 to 2006, and BRFSS 2001 to 2007. RESULTS: Age-adjusted prevalence estimates appeared most similar for NHIS 2005 (physically active: 30.2%, inactive: 40.7%) and NHANES 2005 to 2006 (physically active: 33.5%, inactive: 32.4%). In BRFSS 2005, prevalence of being physically active was 48.3% and inactive was 13.9%. Across all systems, men were more likely to be active than women; non-Hispanic whites were most likely to be active; as age increased, overall prevalence of being active decreased. Prevalence of being active exhibited a significant increasing trend only in BRFSS 2001 to 2007 (P < .001), while prevalence of being inactive decreased significantly in NHANES 1999 to 2006 (P < .001) and BRFSS 2001 to 2007 (P < .001). CONCLUSIONS: Different ways of assessing physical activity in surveillance systems result in different prevalence estimates. Before comparing estimates from different systems, all aspects of data collection and data analysis should be examined to determine if comparisons are appropriate. PMID- 19998847 TI - Television viewing, computer use, and BMI among U.S. children and adolescents. AB - BACKGROUND: To examine the prevalence of television (TV) viewing, computer use, and their combination and associations with demographic characteristics and body mass index (BMI) among U.S. youth. METHODS: The 1999 to 2006 National Health and Nutrition Examination Survey (NHANES) was used. Time spent yesterday sitting and watching television or videos (TV viewing) and using the computer or playing computer games (computer use) were assessed by questionnaire. RESULTS: Prevalence (%) of meeting the U.S. objective for TV viewing (< or =2 hours/day) ranged from 65% to 71%. Prevalence of no computer use (0 hours/day) ranged from 23% to 45%. Non-Hispanic Black youth aged 2 to 15 years were less likely than their non Hispanic White counterparts to meet the objective for TV viewing. Overweight or obese school-age youth were less likely than their normal weight counterparts to meet the objective for TV viewing. CONCLUSIONS: Computer use is prevalent among U.S. youth; more than half of youth used a computer on the previous day. The proportion of youth meeting the U.S. objective for TV viewing is less than the target of 75%. Time spent in sedentary behaviors such as viewing TV may contribute to overweight and obesity among U.S. youth. PMID- 19998848 TI - Healthy people 2010 objectives for physical activity, physical education, and television viewing among adolescents: national trends from the Youth Risk Behavior Surveillance System, 1999-2007. AB - BACKGROUND: To help inform policies and programs, a need exists to understand the extent to which Healthy People 2010 objectives for physical activity, physical education (PE), and television (TV) viewing among adolescents are being achieved. METHODS: As part of the Youth Risk Behavior Surveillance System, 5 national school-based surveys were conducted biennially from 1999 through 2007. Each survey used a 3-stage cross-sectional sample of students in grades 9 to 12 and provided self-reported data from approximately 14,000 students. Logistic regression models that controlled for sex, race/ethnicity, and grade were used to analyze secular trends. RESULTS: During 1999 to 2007, prevalence estimates for regular participation in moderate and vigorous physical activity, participation in daily PE classes, and being physically active in PE classes did not change significantly among female, male, white, black, or Hispanic students. In contrast, the prevalence of TV viewing for 2 or fewer hours on a school day increased significantly among female, male, white, black, and Hispanic students and among students in every grade except 12th grade. CONCLUSIONS: Among US adolescents, no significant progress has been made toward increasing participation in physical activity or school PE classes; however, improvements have been made in reducing TV viewing time. PMID- 19998849 TI - Test-retest repeatability and relative validity of the Global Physical Activity Questionnaire in a developing country context. AB - BACKGROUND: The increasing prevalence of chronic lifestyle diseases in developing countries warrants reliable and valid surveillance of physical activity levels in the population. This study assesses the test-retest repeatability and criterion validity of the WHO-recommended Global Physical Activity Questionnaire (GPAQ) in Vietnamese adults during the dry and wet seasons. METHODS: In 2007 a representative sample of 169 adults (25-64 years) was recruited to determine the GPAQ reliability and validity. GPAQ assesses time and intensity of physical activities spent during a usual week. To assess short and long term reliability, participants completed the GPAQ twice during the dry season 2 weeks apart and again 2 months later during the wet season. For validation purposes, participants wore an accelerometer during the 7 days before the first and last GPAQ assessments. RESULTS: The total GPAQ score showed repeatability correlations of 0.69 after 2 weeks and of 0.55 after 2 months. Total GPAQ score and accelerometer data showed validity correlations of 0.34 and 0.20 in the dry and wet season, respectively. There was a difference in physical activity patterns between the dry and wet seasons. CONCLUSIONS: GPAQ is suitable for surveillance of physical activity among adults in Vietnam. PMID- 19998850 TI - Psychometric properties of the IPAQ: a validation study in a sample of northern Franco-Ontarians. AB - BACKGROUND: The International Physical Activity Questionnaire (IPAQ) has received significant attention since the late 1990s. As it currently stands, its long version has been translated in English, German, Icelandic, Korean, Polish, Spanish, Turkish, and Vietnamese. However no data originating from the self administered long version (last 7 days) of the IPAQ (IPAQ-SALV) is available for French Canadians. This study developed a self-administered long version (last 7 days) of the IPAQ in Canadian French (IPAQ-SALVCF) and assessed its psychometric properties. METHODS: The original IPAQ-SALV was linguistically translated, back translated, and then reviewed in a focus group to ensure its meaning had been retained. Data were collected on a sample of 34 Francophones from Northern Ontario, and the results compared with step counts assessed by 7-day pedometer recording. Test-retest reliability was examined with a 24-hour delay between questionnaire completion on day 8 and day 9 of the protocol. Convergent validity was assessed by comparing IPAQ-SALVCF (last 7 days) results to average step counts over a 7-day period. RESULTS: Intraclass correlation coefficients (ICC) revealed that the IPAQ-SALVCF results were stable between days. The ICC for total activity scores was highest at 0.93 (CI: 0.86 to 0.97). Total activity scores were also significantly related to pedometer step counts (Pearson r = .66 P < .01). These results confirm those obtained in prior research. CONCLUSION: The IPAQ-SALVCF is a reliable and valid measure of physical activity for French Canadians. PMID- 19998851 TI - The work and home activities questionnaire: energy expenditure estimates and association with percent body fat. AB - BACKGROUND: Understanding and increasing physical activity requires assessment of occupational, home, leisure and sedentary activities. METHODS: A physical activity questionnaire was developed using data from a large representative U.S. sample; includes occupational, leisure and home-based domains; and produces estimates of energy expenditure, percent body fat, minutes in various domains, and meeting recommendations. It was tested in 396 persons, mean age 44 years. Estimates were evaluated in relation to percent body fat measured by dual-energy x-ray absorptiometry. RESULTS: Median energy expenditure was 2365 kcal (women) and 2960 kcal (men). Women spent 35.1 minutes/day in moderate household activities, 13.0 minutes in moderate leisure and 4.0 minutes in vigorous activities. Men spent 18.0, 22.5 and 15.6 minutes/day in those activities, respectively. Men and women spent 276.4 and 257.0 minutes/day in sedentary activities. Respondents who met recommendations through vigorous activities had significantly lower percent body fat than those who did not, while meeting recommendations only through moderate activities was not associated with percent body fat. Predicted and observed percent body fat correlated at r = .73 and r = .82 for men and women respectively, P < .0001. CONCLUSIONS: This questionnaire may be useful for understanding health effects of different components of activity, and for interventions to increase activity levels. PMID- 19998852 TI - Objective measures of the environment and physical activity--results of the environment and physical activity study in English adults. AB - BACKGROUND: Physical activity has been positively associated with a range of objectively measured environmental variables. We explored the relationship of walking and other categories of physical activity with objectively measured activity specific environmental variables in a U.K. population. METHODS: We used a geographical information system (GIS) and gender specific multivariate models to relate 13,927 participants' reported levels of physical activity with a range of measures of the environment. RESULTS: Access to green space and area levels of crime were not associated with walking for recreation. Distance to facilities had either no or only a small effect on the uptake of different activities. Odds ratios of cycling for leisure dropped as local traffic density increased for both genders. Compared with the lowest quartile for traffic density the likelihood of reporting any cycling for leisure was OR 0.42, (95% CI 0.32 to 0.52, P < .001) for women and OR 0.41, (95% CI 0.33 to 0.50, P < .001) for men in the highest quartile. CONCLUSIONS: We were unable to reproduce results observed in previous studies. Future research should use large representative population samples from multiple areas to maximize environmental variability and if feasible use both objective and subjective measures of physical activity and the environment. PMID- 19998853 TI - Measuring walking for physical activity surveillance--the effect of prompts and respondents' interpretation of walking in a leisure-time survey. AB - BACKGROUND: Walking is the most prevalent form of leisure time physical activity (LTPA). Advances in measurement of walking depend on understanding sources of error in self report. We examined the effect of prompting "walking for exercise, recreation, and sport" (WERS) upon surveillance estimates of LTPA and assessed what types of walking were recalled when reporting LTPA generally and when WERS was prompted specifically. METHODS: Data were collected by telephone survey from a random sample of 3,415 Australian adults (> or = 15 yrs). Respondents were asked first to recall any type of LTPA they participated in (unprompted) and if walking was not mentioned, WERS was prompted. All walkers were asked to describe the type of walking they did. Open-ended responses were categorized according to physical activity measurement dimensions. RESULTS: Forty three percent did not report WERS unless prompted to do so. The prevalence of meeting recommendations by all LTPA was reduced by 10% for both genders and across all age groups if not prompted to recall WERS. The interpretation of WERS was broad and included travel related walking and dog walking whether unprompted or prompted. CONCLUSIONS: Current challenges in walking surveillance include ensuring that both researchers and respondents understand WERS in a standardized manner. PMID- 19998854 TI - Observing children's playground activity levels at 13 Illawarra primary schools using CAST2. AB - BACKGROUND: Declining levels of children's physical activity may contribute to Australia's increasing childhood obesity epidemic. School recess is an underutilized opportunity to increase children's physical activity. METHODS: Thirteen regional Australian public primary schools participated in the study (2946 children). The Children's Activity Scanning Tool 2 (CAST2) collected observational playground physical activity data. The research also addressed: length of break, socioeconomic status (SES), gender, number of scanning days, and instrument calibration. RESULTS: The proportions of Moderate or Vigorous Physically Activity (MVPA) children at the observed schools ranged from 0.4 to 0.7. The odds ratio of boys being MVPA relative to girls ranged from 0.8581 to 2.137. There were significant differences between the mean proportions of 3 days of activity (range P = .001 to P = .015) and no association between SES school groupings (deviance ratio: 0.48; P = .503). Interrater reliability for instrument calibration using Spearman correlations coefficients ranged from r = .71 to r = .99. CONCLUSIONS: There were significant differences between proportions of MVPA children at the 13 schools and between male and female populations. There was no association between playground physical activity and SES. The monitoring period for CAST2 should be at least 3 days. Interrater reliability indicates that correlations between observers were consistently high. PMID- 19998855 TI - Physical activity in the park setting (PA-PS) questionnaire: reliability in a California statewide sample. AB - BACKGROUND: The Physical Activity in Parks Setting (PA-PS) instrument is a series of survey questions designed by a consortium of public health and leisure research scholars to gauge park-based physical activity for use in civilian, noninstitutionalized populations. This paper introduces this self-reported instrument and provides test-retest reliability results. METHODS: Data to test the instrument reliability were collected during 2 waves in 2008 through the California Outdoor Recreation Opinions and Attitudes Telephone Survey. To conduct test-retest reliability we examined the agreement between 100 randomly reselected respondents from the first wave of respondents (n=2004) that answered the same survey within 21 to 30 days of the initial administration. RESULTS: The reliability of measures that categorized individual park use and visitation with others provided moderate levels of agreement (Kappa = 0.44 to 0.64). Questions about park features, facilities and amenity use, and specific park-based physical activity participation were of fair to substantial agreement (Kappa = 0.21 to 0.90) depending on the item in question. CONCLUSION: The results from these test retest reliability analyses suggest the PA-PS items were reliable and should be considered in future population surveys that assess park visitation patterns and park-based physical activity levels. PMID- 19998856 TI - Measuring physical activity self-regulation strategies in older adults. AB - BACKGROUND: Theoretically, self-regulatory strategies (e.g., goal setting, self monitoring) are an important influence of behavior change, but very little research has examined the relationship between self-regulation and physical activity (PA) behavior. Petosa's (1993) 43-item PA self-regulation scale (PASR 43) affords the opportunity for studying this construct in the context of PA; however the PASR-43 has not been tested for structural aspects of validity. Therefore, this study examines the structural validity of the PASR-43 in older adults. METHODS: The structural validity of the PASR-43 was tested in a large sample of older adults from North and South Carolina and Ohio (N = 460) using maximum likelihood estimation and confirmatory factor analysis in AMOS 5.0. RESULTS: The original 6-factor model for the PASR-43 scale did not represent an acceptable fit to the data (chi2 = 4732.25, df = 845, P < .0001, RMSEA = 0.10, NNFI = 0.67, CFI = 0.71). Based on a post hoc specification search, iterative model modifications resulted in a 12-item PA self-regulation scale (PASR-12) that represented an excellent fit to the data (chi2 = 70.75, df = 39, P = .001, RMSEA = 0.04, NNFI = 0.98, CFI = 0.99). CONCLUSIONS: The PASR-12 provides a concise and valid measure of PA self-regulation for use with older adults. Future studies should cross-validate the PASR-12 and examine invariance across time and between age, ethnic, gender, and geographical groups. PMID- 19998857 TI - Validation of the Neighborhood Environment Walkability Scale (NEWS) items using geographic information systems. AB - BACKGROUND: Concurrent validity of Neighborhood Environment Walkability Scale (NEWS) items was evaluated with objective measures of the built environment using geographic information systems (GIS). METHODS: A sample of 878 parents of children 10 to 16 years old (mean age 43.5 years, SD = 6.8, 34.8% non-White, 63.8% overweight) completed NEWS and the International Physical Activity Questionnaire. GIS was used to develop 1-mile street network buffers around participants' residences. GIS measures of the built environment within participants' buffers included percent of commercial and institutional land uses; number of schools and colleges, recreational facilities, parks, transit stops, and trees; land topography; and traffic congestion. RESULTS: Except for trees and traffic, concordance between the NEWS and GIS measures were significant, with weak to moderate effect sizes (r = -0.09 to -0.36, all P < or = 01). After participants were stratified by physical activity level, stronger concordance was observed among active participants for some measures. A sensitivity analysis of self-reported distance to 15 neighborhood destinations found a 20-minute (compared with 10- or 30-minute) walking threshold generally had the strongest correlations with GIS measures. CONCLUSIONS: These findings provide evidence of the concurrent validity of self-reported built environment items with objective measures. Physically active adults may be more knowledgeable about their neighborhood characteristics. PMID- 19998858 TI - Validation of the Jackson Heart Study Physical Activity Survey in African Americans. AB - BACKGROUND: Physical inactivity has been consistently linked to cardiovascular disease, yet few instruments have been validated for assessment of physical activity in African Americans, a group particularly vulnerable to heart disease. The current study aimed to establish the psychometric properties of the activity survey used in the Jackson Heart Study (JHS) among African Americans, the JHS Physical Activity Cohort survey (JPAC). METHODS: Test-retest reliability over 2 weeks was assessed using a convenience sample of 40 African Americans. Convergent validity with accelerometer and pedometer data were assessed in 2 samples from the JHS (N = 404 and 294, respectively). RESULTS: Test-retest reliability was excellent, with intraclass correlations = .99 for the JPAC total and index scores. Higher JPAC total scores were significantly associated with higher raw accelerometer and pedometer counts. Spearman correlations between JPAC total scores and accelerometer (rho = .24) and pedometer counts (rho = .32) were consistent with these results. Most subscales were significantly correlated with the objective measures. The JPAC total score was most strongly associated with objectively-measured activity. CONCLUSION: This study provides support for the reliability and validity of the JPAC as a tool for assessing physical activity among African Americans across a variety of domains. PMID- 19998859 TI - Actigraph calibration in obese/overweight and type 2 diabetes mellitus middle aged to old adult patients. AB - BACKGROUND: Several methods exist to asses and control physical intensity levels of subjects engaged in physical activities programs, accelerometry is a method that could be easily used in the field. The purposes were: to calibrate Actigraph in middle-aged to old obese/overweight and DM2 adult patients; and to determine the threshold counts for sedentary, light, moderate, and vigorous physical activity (PA). METHODS: Sample comprise 26 participants (62.6 +/- 6.5 years of age) of both gender. Counts and VO2 were simultaneously assessed during: resting, seating, standing, walking at 2.5 km x h(-1), 5 km x h(-1), and 6 km x h(-1). A hierarchical linear model was used to derive a regression equation between MET and counts. Receiver operating characteristics (ROC) analysis was used to define thresholds for PA levels. RESULTS: The regression equation was: MET 1.388400490262 + 0.001312683420044 (counts x min(-1)), r = .867. The threshold counts for sedentary-light, light-moderate and moderate-vigorous PA were: 200, 1240, 2400 counts-min(-1) respectively. CONCLUSION: The Actigraph is a valid and useful device for the assessment of the amount of time spent in each PA intensity levels in obese/overweight and DM2 middle-aged to old adult patients. PMID- 19998860 TI - Comparison of Kenz Lifecorder versus actigraph physical activity output in free living conditions. AB - BACKGROUND: The Kenz Lifecorder EX (KL) is a relatively new, moderately priced, user friendly accelerometer that tracks step counts and time spent in various intensity classifications. Thus, the KL is an attractive instrument for researchers and the public. However, there is limited research comparing the KL's output to other accelerometers during free-living conditions. Therefore the purpose of this study was to compare KL versus ActiGraph (AG) outputs of step counts and time spent in various intensity classifications during free-living conditions. METHODS: Ten men and 10 women volunteers wore an AG (right side) and 2 KL (right side: KL-R vs. left side: KL-L) accelerometers on their waistline during waking hours for one day. RESULTS: KL-R vs. KL-L yielded similar physical activity (PA) output. The AG recorded fewer steps compared with KL-L (P = .002) but was similar to the KL-R. The KL-R and KL-L yielded lower estimates of accumulated time spent in moderate PA compared with most AG intensity derivations (P < .003). There were no differences between KL-R and KL-L vs. the AG for time spent in vigorous PA. CONCLUSIONS: The KL provides similar estimates of step counts and time spent in vigorous PA compared with established AG intensity derivations. PMID- 19998861 TI - Compliance with wearing physical activity accelerometers in high school students. AB - BACKGROUND: Accelerometer use in physical activity research has become increasingly popular but is prone to problems with missing data, which complicate the data reduction and analysis process. The purpose of this study was to determine the effect of hypothesized compliance strategies on improving compliance with wearing a physical activity accelerometer in high school students. METHODS: Each of four local high schools was assigned to one of four compliance strategies: (1) receiving three phone calls, (2) completing a daily journal, (3) compensation contingent on number of complete (> or = 10 hours) days of data, and (4) control condition. Participants wore ActiGraph accelerometers for seven days to determine compliance and physical activity. RESULTS: The contingent group had the highest level of compliance with 96% of the participants acquiring at least four of seven complete days of data. After controlling for grade level, school level percent minority students, and school level socioeconomic status (SES), the contingent group's compliance remained significantly higher (P = .04) than the journal (85%), phone (72%), and control (70%) participants. CONCLUSIONS: The contingent compliance strategy improved the amount of time the students wore the monitor each day and, thus, the total number of days with > or =10 hours of data. PMID- 19998862 TI - Dietary management of moderate malnutrition: time for a change. PMID- 19998863 TI - Proposed recommended nutrient densities for moderately malnourished children. AB - Recommended Nutrient Intakes (RNIs) are set for healthy individuals living in clean environments. There are no generally accepted RNIs for those with moderate malnutrition, wasting, and stunting, who live in poor environments. Two sets of recommendations are made for the dietary intake of 30 essential nutrients in children with moderate malnutrition who require accelerated growth to regain normality: first, for those moderately malnourished children who will receive specially formulated foods and diets; and second, for those who are to take mixtures of locally available foods over a longer-term to treat or prevent moderate stunting and wasting. Because of the change in definition of severe malnutrition, much of the older literature is pertinent to the moderately wasted or stunted child. A factorial approach has been used in deriving the recommendations for both functional, protective nutrients (type I) and growth nutrients (type II). PMID- 19998864 TI - Choice of foods and ingredients for moderately malnourished children 6 months to 5 years of age. AB - There is consensus on how to treat severe malnutrition, but there is no agreement on the most cost-effective way to treat infants and young children with moderate malnutrition who consume cereal-dominated diets. The aim of this review is to give an overview of the nutritional qualities of relevant foods and ingredients in relation to the nutritional needs of children with moderate malnutrition and to identify research needs. The following general aspects are covered: energy density, macronutrient content and quality, minerals and vitamins, bioactive substances, antinutritional factors, and food processing. The nutritional values of the main food groups--cereals, legumes, pulses, roots, vegetables, fruits, and animal foods--are discussed. The special beneficial qualities of animal-source foods, which contain high levels of minerals important for growth, high-quality protein, and no antinutrients or fibers, are emphasized. In cereal-dominated diets, the plant foods should be processed to reduce the contents of antinutrients and fibers. Provision of a high fat content to increase energy density is emphasized; however, the content of micronutrients should also be increased to maintain nutrient density. The source of fat should be selected to supply optimal amounts of polyunsaturated fatty acids (PUFAs), especially n-3 fatty acids. Among multiple research needs, the following are highlighted: to identify the minimum quantity of animal foods needed to support acceptable child growth and development, to examine the nutritional gains of reducing contents of antinutrients and fibers in cereal- and legume-based diets, and to examine the role of fat quality, especially PUFA content and ratios, in children with moderate malnutrition. PMID- 19998865 TI - Dietary counseling in the management of moderate malnourishment in children. AB - BACKGROUND: Dietary counseling is an integral part of treating malnutrition. A first step toward improving the management of moderate malnutrition is to evaluate dietary messages in current programs and assess their adequacy and effectiveness. OBJECTIVES: To ascertain current recommendations regarding family foods for the treatment of moderate malnutrition and assess whether these are likely to meet nutritional requirements for rehabilitation; to review the effectiveness of dietary counseling in the management of moderate malnutrition. METHODS: Information was requested from 10 United Nations agencies or donors, 20 international nongovernmental organizations, 3 pediatric associations, and 6 national programs about the dietary advice they give to caregivers of moderately malnourished children. Adequacy was assessed by comparing dietary recommendations with nutritional requirements. Linear programming was used to identify problem nutrients. A literature search was conducted of studies using family foods for rehabilitating malnourished children. RESULTS: There was a greater emphasis on providing food supplements for rehabilitation than on utilizing family foods. Dietary recommendations were mostly vague and unlikely to be effective. Those developed by the World Health Organization and the Food and Agriculture Organization for well-nourished children may meet nutritional requirements in moderate malnutrition if the recommendations are made more prescriptive. Zinc and vitamin E emerged as possible problem nutrients. Intervention studies in wasted children suggest that counseling caregivers about family foods can achieve good rates of weight gain. CONCLUSIONS: Dietary counseling can be effective in managing malnutrition, but it is often weak or absent and should be strengthened. More attention will need to be given to formulating the messages and improving counseling skills. PMID- 19998866 TI - Current and potential role of specially formulated foods and food supplements for preventing malnutrition among 6- to 23-month-old children and for treating moderate malnutrition among 6- to 59-month-old children. AB - Reducing child malnutrition requires nutritious food, breastfeeding, improved hygiene, health services, and (prenatal) care. Poverty and food insecurity seriously constrain the accessibility of nutritious diets that have high protein quality, adequate micronutrient content and bioavailability, macrominerals and essential fatty acids, low antinutrient content, and high nutrient density. Diets based largely on plant sources with few animal-source and fortified foods do not meet these requirements and need to be improved by processing (dehulling, germinating, fermenting), fortification, and adding animal-source foods, e.g., milk, or other specific nutrients. Options include using specially formulated foods (fortified blended foods, commercial infant cereals, or ready-to-use foods [RUFs; pastes, compressed bars, or biscuits]) or complementary food supplements (micronutrient powders or powdered complementary food supplements containing micronutrients, protein, amino acids, and/or enzymes or lipid-based nutrient supplements (120 to 250 kcal/day), typically containing milk powder, high-quality vegetable oil, peanut paste, sugar, and micronutrients. Most supplementary feeding programs for moderately malnourished children supply fortified blended foods, such as corn-soy blend, with oil and sugar, which have shortcomings, including too many antinutrients, no milk (important for growth), suboptimal micronutrient content, high bulk, and high viscosity. Thus, for feeding young or malnourished children, fortified blended foods need to be improved or replaced. Based on success with ready-to-use therapeutic foods (RUTFs) for treating severe acute malnutrition, modifying these recipes is also considered. Commodities for reducing child malnutrition should be chosen on the basis of nutritional needs, program circumstances, availability of commodities, and likelihood of impact. Data are urgently required to compare the impact of new or modified commodities with that of current fortified blended foods and of RUTF developed for treating severe acute malnutrition. PMID- 19998867 TI - Proceedings of the World Health Organization/ UNICEF/World Food Programme/United Nations High Commissioner for Refugees Consultation on the management of moderate malnutrition in children under 5 years of age. PMID- 19998868 TI - Slow wave sleep: does it matter? PMID- 19998870 TI - Slow wave homeostasis and synaptic plasticity. PMID- 19998869 TI - Regulation and functional correlates of slow wave sleep. PMID- 19998871 TI - The role of slow wave sleep in memory processing. PMID- 19998873 TI - In the pink. PMID- 19998872 TI - Enhancement of slow wave sleep: implications for insomnia. PMID- 19998874 TI - From employee to self-employed. PMID- 19998875 TI - Low back pain in women. PMID- 19998876 TI - Self-service appointments. PMID- 19998877 TI - Patient information. Planning or avoiding pregnancy naturally. PMID- 19998878 TI - Mixed signals. Treating overlapping symptoms of urinary incontinence. PMID- 19998879 TI - Transition to depression? Perimenopause, menopause and mental health. PMID- 19998880 TI - Aging gracefully. Skin care for midlife and older women. PMID- 19998881 TI - Women's health care in a spa setting. An inviting intersection of disciplines. PMID- 19998882 TI - Mammography and biopsy. An overview for primary care providers. PMID- 19998883 TI - HPV in adolescents. Making the wake-up call. PMID- 19998884 TI - Germ warfare. Probiotics restore peace. PMID- 19998885 TI - Stop, look, listen. Acute coronary syndrome in women. PMID- 19998886 TI - Heading off VTE. The role of inherited coagulation disorders. PMID- 19998887 TI - Preparing for the perfect storm. Disaster planning for older adults. PMID- 19998888 TI - Don't have a PDA yet? Consider these points. PMID- 19998889 TI - Behind the headlines. PMID- 19998891 TI - Get recognized! PMID- 19998890 TI - Pennsylvania 'unleashes' NPs leaders credit governor's interest. PMID- 19998892 TI - Review of informed consent. PMID- 19998893 TI - Where to go to be in the know. PMID- 19998894 TI - Icteric skin, sclera and mucous membranes. PMID- 19998895 TI - Foreign body extraction. PMID- 19998896 TI - Patient information. Skin rejuvenation: a guide. PMID- 19998897 TI - The long and winding road. Arriving at safe medication management in LTC setting. PMID- 19998898 TI - Perioperative concerns in older adults: multiple risks require attention. PMID- 19998900 TI - Taming the flame. Strategies for GERD and heartburn. PMID- 19998899 TI - Reaching out to victims of sexual assault. SANE training adds dimension to NP practice. PMID- 19998901 TI - Beneficence, prescriptions and the nurse practitioner. An ethics case study. PMID- 19998902 TI - Rusty hinges. Understanding osteoarthritis pain. PMID- 19998904 TI - Difficult patients, Part 2. More advice for handling those headaches. PMID- 19998903 TI - Relating to research. Sifting through data to cultivate true evidence-based practice. PMID- 19998906 TI - Stop the train. PMID- 19998905 TI - The last ride. PMID- 19998907 TI - A guide to clinical privileges. PMID- 19998908 TI - Cough, fever and weight loss. PMID- 19998909 TI - Subfertility in men. PMID- 19998910 TI - Patient information. Cranberries: small fruit, big health benefit. PMID- 19998911 TI - The lungs have it. Inhaled insulin therapy. PMID- 19998912 TI - Diabetes in older adults. Consider presentation, functioning and more. PMID- 19998913 TI - Altering the course. Screening for prediabetes. PMID- 19998914 TI - Intrauterine contraception. Considerations for special populations...and more. PMID- 19998915 TI - Good nutrition on the go. Eating healthy is a family affair. PMID- 19998916 TI - Real solutions for real life. Smoking cessation in today's busy practice settings. PMID- 19998917 TI - It's time to speak up. PMID- 19998918 TI - A cry for help. PMID- 19998919 TI - All cash, no office. PMID- 19998920 TI - Diabetes in minority men. PMID- 19998921 TI - Insuring your aesthetics practice. PMID- 19998922 TI - Hemorrhoid evacuation. PMID- 19998923 TI - Patient information. Determining your cardiometabolic health. PMID- 19998924 TI - Toward better clinical asthma management. The nurse practitioner as asthma educator. PMID- 19998925 TI - Childhood depression. Diagnosis and management in an era of black box warnings. PMID- 19998926 TI - Cytolytic vaginosis and lactobacillosis. Consider these conditions with all vaginosis symptoms. PMID- 19998927 TI - Neurostabilizers for migraine prevention. From dosage administration to patient expectations. PMID- 19998928 TI - Primary prevention of stroke. Recognition of risk factors. PMID- 19998929 TI - When should older adults stop driving? PMID- 19998930 TI - More than a workplace danger. An update on latex sensitivity. PMID- 19998931 TI - Adults with developmental disabilities. Guidance for quality clinical care. PMID- 19998932 TI - First things first. Your new role as an NP. PMID- 19998933 TI - Cherry Ames, where are you? PMID- 19998934 TI - This prescription should be nationwide. PMID- 19998935 TI - Playing by the rule book. PMID- 19998936 TI - Heading off liability claims. PMID- 19998937 TI - Digital nerve block. PMID- 19998938 TI - Patient information. Cranial abnormalities. PMID- 19998939 TI - Dysfunctional elimination syndrome. Where constipation, daytime urinary problems and bedwetting merge. PMID- 19998940 TI - Preventing obesity in young children. Where theory meets practice. PMID- 19998941 TI - The piercing truth. Tattoos and body piercing in adolescents. PMID- 19998942 TI - Work-related low back pain. An overview of key clinical concepts. PMID- 19998943 TI - Interstitial cystitis. a concise guide. PMID- 19998944 TI - Paradigm shift. New testing guidelines for HIV. PMID- 19998945 TI - Heart failure. A common sense approach. PMID- 19998946 TI - Always a threat. Legionellosis disease. PMID- 19998947 TI - Advances in adult immunization. Time to renew vaccination efforts. PMID- 19998948 TI - Contract negotiation. More than money is at stake. PMID- 19998949 TI - Polish your resume. Moving from RN to NP. PMID- 19998950 TI - Standing strong. PMID- 19998951 TI - Standing strong: update on the Gold Street Clinic. PMID- 19998952 TI - Sex differences in sleep. PMID- 19998953 TI - Electronic prescribing. PMID- 19998955 TI - Patient information. Preventing and treating pressure sores. PMID- 19998954 TI - In between the shoulder blades and over the maxilla. PMID- 19998956 TI - The asthma-allergy conncection. Complex companions. PMID- 19998957 TI - Asthma's predecessor: atopic dermatitis. PMID- 19998958 TI - Hereditary hemorrhagic telangiectasia. Below the surface, health dangers threaten. PMID- 19998959 TI - Lasers, peels, injections and masks. An overview of treatment modalities for scars. PMID- 19998960 TI - Newborn screening. As testing choices expand, so must your knowledge. PMID- 19998961 TI - Falls by older adults. Think preventable, not unavoidable. PMID- 19998962 TI - Overcoming clinical inertia in diabetes care. Point-of-care A(1C) testing a valuable tool. PMID- 19998963 TI - Preventing cervical cancer. The age of vaccination has arrived. PMID- 19998964 TI - Stress testing to evaluate chest pain. Clear guidance for cloudy issues. PMID- 19998965 TI - What color is your sugarbowl? A closer look at sweeteners. PMID- 19998966 TI - Applying for the job. Tips for savvy communicators. PMID- 19998967 TI - Learning from those at risk. PMID- 19998968 TI - Wake-up call in Oregon. Challenge averted at session's end. PMID- 19998969 TI - Osteoporosis in men: a real threat. PMID- 19998970 TI - Welcome to aesthetics: the good, the bad and the ugly. PMID- 19998971 TI - Sebaceous cyst removal. PMID- 19998972 TI - Patient information. Diabetes and meal planning. PMID- 19998973 TI - Hypertension and renal disease risk. Eyes on the end result. PMID- 19998974 TI - Pelvic organ prolapse. A new option offers effectiveness and ease of use. PMID- 19998975 TI - Breakthrough breastfeeding strategies. Practical advice for time-pressed NPs. PMID- 19998976 TI - Never too old for HIV. Reducing risk in your patients. PMID- 19998977 TI - Community-associated MRSA. Tools for treatment and prevention. PMID- 19998978 TI - Complex regional pain syndrome. The role of spinal cord stimulation. PMID- 19998979 TI - Unforgettable. Responding to PTSD in primary care. PMID- 19998980 TI - An overview of prostatitis. Compliance can be biggest challenge. PMID- 19998981 TI - Need malpractice insurance? Get in line. PMID- 19998982 TI - Defend and conquer. Protect you practice's accounts receivable. PMID- 19998983 TI - Face time. Making the most of legislative meetings. PMID- 19998984 TI - Difficult patients. Advice for handling those headaches. PMID- 19998985 TI - Dance as if no one were watching. PMID- 19998986 TI - Make prenatal depression screening a priority. PMID- 19998987 TI - Writing to get the grant. PMID- 19998989 TI - Communicating by e-mail. PMID- 19998988 TI - Sex differences in osteoarthritis. PMID- 19998991 TI - Patient information. Managing osteoarthritis pain. PMID- 19998990 TI - Cough... followed by calf pain. PMID- 19998992 TI - Osteomyelitis: principles to guide prevention, diagnosis and treatment. PMID- 19998993 TI - Wound closure techniques: key concepts. PMID- 19998994 TI - Smoking cessation. A protocol for adolescents. PMID- 19998995 TI - Glucose meters. A guide for you and your patients. PMID- 19998996 TI - Hereditary colorectal cancer syndromes. Start risk assessment in primary care. PMID- 19998997 TI - Fading away. Caregiver stress in spouses of patients with Alzheimer's disease. PMID- 19998998 TI - Inflammatory bowel disease. A clinical review of Crohn's disease and ulcerative colitis. PMID- 19998999 TI - Mental health and pregnancy. The link between prenatal and postpartum depression. PMID- 19999000 TI - Finding a mentor. You need more than one. PMID- 19999001 TI - Pregnancy and disability. PMID- 19999002 TI - An Office of Men's Health: still waiting. PMID- 19999003 TI - When to call the BON. PMID- 19999004 TI - HIPAA privacy complaints. PMID- 19999005 TI - Orotracheal intubation. PMID- 19999006 TI - Patient information. Ingrown toenail. PMID- 19999007 TI - Going the distance. New pain treatment provide longer stronger relief. PMID- 19999008 TI - Psoriasis in adolescents. Psychosocial effects warrant close attention. PMID- 19999009 TI - Antidepressant treatment for fibromyalgia. Avoiding two dangerous syndromes. PMID- 19999010 TI - Anemia in the older adult. Engineering George's road to better health. PMID- 19999011 TI - Achieving lipid goals. A study of outcomes in an NP Clinic. PMID- 19999012 TI - On the trail of a killer. Assessing suicide risk in men. PMID- 19999013 TI - The nuts and bolts. 10 tips for starting your own business. PMID- 19999014 TI - Working a conference. Be productive and have fun! PMID- 19999015 TI - Empowerment through dignity. PMID- 19999016 TI - [Guide for diagnosis and treatment of asthma]. PMID- 19999017 TI - [Efficacy of two methods of administration of salbutamol-ipratropium bromide in asthma crises]. AB - BACKGROUND: Asthma acute crisis is one of the most frequent pediatric emergencies. Quick treatment with an efficient inhaled therapy may diminish considerably the number of hospitalizations and time of crises. OBJECTIVE: To compare the efficacy of administration of salbutamol and ipratropium bromide by nebulization with supplementary oxygen vs salbutamol and ipratropium bromide administered in device of measured doses with space chamber for treatment of asthmatic crises in children. PATIENTS AND METHOD: 45 children from 1 to 12 years of age were randomly selected; they were divided into two groups and were classified according to respiratory difficulty degree. Then inhaled therapy proposed was administered three times, and oximetry and hospitalization criteria were determined. RESULTS: Pediatric patients with non-severe asthma crisis had a better course with salbutamol and ipratropium bromide combination administered by device of measures doses and space chamber than with the same drugs nebulized with supplementary oxygen, and percentage of hospitalizations of children with asthma crises was reduced (p = 0.037). CONCLUSION: Administration of salbutamol and ipratropium bromide with device of measured doses was more efficient that their administration nebulized with supplementary oxygen for treating asthmatic crises in children. PMID- 19999018 TI - [The role of acupuncture in asthma]. AB - Asthma is a chronic inflammatory syndrome of airways, in which different cells and their mediators play a role originating damage clinically expressed by recurrent respiratory symptoms. Current therapy of asthma is focused to reduce airway inflammation, diminishing long-term bronchial hyperactivity and rescue treatment (when required). Unfortunately, the failure of current therapies of asthma has increased the number of patients searching for approximations in complementary and alternative medicine. Scientific evidence is still scarce for establishing that alternative medicine is more effective than placebo in asthma. However, this does not mean that it is ineffective, maybe more multicentric clinical assays are needed, with methodological rigor, to establish final conclusions of efficacy and safety of complementary medicine. Thus, until such results are available, it is better to be skeptical and not to recommend acupuncture for asthma treatment. PMID- 19999019 TI - [Risk factors of food allergy]. AB - Food allergy or allergic food hypersensitivity is defined as an adverse immunologic reaction caused by immunologic mechanisms mediated or not by IgE. It is a complex disease influenced by polygenetic heritance and environmental factors. Many risk factors have been investigated, pre natal and post natal, and variable and controversial results have been obtained. The most important risk factors associated with food allergy are atopy, lack of breast feeding at least three to six months and early weaning (before four-six months). PMID- 19999020 TI - [Chronic granulomatous disease]. AB - Chronic granulomatous disease (CGD) is a primary immunodeficiency, a phagocyte defect that appears in 1:200,000 live births and is produced by mutations in the genes that codify for the enzyme nicotinamide adenine dinucleotide phosphate oxidase (NADPH oxidase). The inheritance form is X linked (> 60%) or autosomic recesive (30-40%). The NADPH oxidase is responsible for the production of reactive oxygen species (ROS) in the activated phagocyte ("respiratory burst"). When present, mutations on the NAPDH oxidase genes do not allow the ROS production, making the neutrophils of these patients incapable to destroy pathogens. These patients are especially susceptible to infections by staphylococcus, fungi and some gram-negative bacteria. The main clinical manifestations include recurrent life-threatening episodes of lymphadenitis, abscess, pneumonias, osteomyelitis, granuloma formation and sepsis. The diagnosis is suggested by a history of recurrent infections, familiar cases, fail to grow and confirmed with an altered test of ROS production and the specific mutation. Allogenic stem cells transplant is the curative treatment. The early diagnosis and the treatment with prophylactic antibiotics and interferon-gamma have modified favorably the morbidity and mortality of these patients. PMID- 19999022 TI - [Efficacy of acupuncture in asthma]. PMID- 19999021 TI - [Spontaneous pneumomediastinum related to near-fatal asthma. A report of a case]. AB - Spontaneous pneumomediastinum is an infrequent condition in children under 18 years old, of various etiologies, and benign clinical course. Acute asthma is among the main reasons for medical care in the emergency room, but there is controversy in defining near-fatal asthma. We present the case of an adolescent with chronic severe asthma and risk factors for near-fatal asthma, followed by the presence of pneumomediastinum and subcutaneous emphysema. The diagnosis was made with clinical data and supported by proper radiographic studies. Both pneumomediastinum as well as near-fatal asthma are two infrequent conditions for asthma. Early detection of both results in good patient care. PMID- 19999023 TI - One by one. PMID- 19999024 TI - A rewarding mission. PMID- 19999025 TI - NPs as hospitalists. An evolving role. PMID- 19999026 TI - Lethargy in a 9-year-old girl. A good history goes a long way. PMID- 19999027 TI - On corporations and contracts. Starting a business, sudden change in employment terms. PMID- 19999028 TI - Navigating rough terrain. Steering around the rocky road ot reimbursement. PMID- 19999029 TI - Patient information: bioidentical hormone therapy. Could it be right for you? PMID- 19999030 TI - Malnutrition in older adults. Deciphering a complex syndrome. PMID- 19999031 TI - Hormone therapy in menopause. Exploring current recommendations. PMID- 19999032 TI - Atopic dermatitis. Stop the march to allergy and beyond. PMID- 19999033 TI - Partners for better athma control. Newest guidelines highlight value of NP care. PMID- 19999034 TI - More fish, more nuts, more seed oils. Omega fatty acids and your patients' health. PMID- 19999035 TI - Prevention with power. Breaking through barriers to pneumococcal vaccination. PMID- 19999036 TI - Bouncing back after failure. Don't be so hard on yourself! PMID- 19999037 TI - The value of a mentor. There's always something more to learn. PMID- 19999038 TI - [Setting limit values for chemical substances in the workplace: DNEL(INH) setting according to REACH principles following the example of 2-butyne-1,4-diol]. AB - BACKGROUND: Derived No Effect Level (DNEL(inh)) has been set for occupational exposure to but-2-yno-1,4-diol according to REACH principles. Maximum allowable concentration (MAC) and DNEL(inh) have been compared. MATERIAL AND METHODS: Experimental data from two inhalation studies on rats and three oral studies have been used to calculate DNEL. RESULTS: Estimated DNEL(inh) values show significant differences and fall within the range of 0.33-0.02 mg/m3, depending on the chosen experiment and critical effect. It seems that a 30-day inhalation study best reflects the penetration of xenobiotic to the human organism. This experiment has been used to set MAC value of but-2-yno-1,4-diol of 0,25 mg/m3 and it is close to local DNEL(inh)--0.10 mg/m3. Both values have been estimated on the same starting point NOAEC(loc)--0.5 mg/m3, but different assessment factors have been applied. CONCLUSIONS: A general feeling is that because of the differences in methodologies for calculating DNELs versus those used for calculating health based OELs, the DNEL will tend to be lower than any corresponding health-based OEL for that chemical. This indicates that the OEL does not provide the appropriate level of protection required by REACH. The calculation leads to a new value (DNEL) that requires different risk management measures and operational conditions. PMID- 19999039 TI - [Upper extremities, neck and back symptoms in office employees working at computer stations]. AB - BACKGROUND: To obtain current data on the occurrence ofwork-related symptoms of office computer users in Poland we implemented a questionnaire survey. Its goal was to assess the prevalence and intensity of symptoms of upper extremities, neck and back in office workers who use computers on a regular basis, and to find out if the occurrence of symptoms depends on the duration of computer use and other work-related factors. MATERIAL AND METHODS: Office workers in two towns (Warszawa and Katowice), employed in large social services companies, were invited to fill in the Polish version of Nordic Questionnaire. The questions included work history and history of last-week symptoms of pain of hand/wrist, elbow, arm, neck and upper and lower back (occurrence and intensity measured by visual scale). Altogether 477 men and women returned the completed questionnaires. Between-group symptom differences (chi-square test) were verified by multivariate analysis (GLM). RESULTS: The prevalence of symptoms in individual body parts was as follows: neck, 55.6%; arm, 26.9%; elbow, 13.3%; wrist/hand, 29.9%; upper back, 49.6%; and lower back, 50.1%. Multivariate analysis confirmed the effect of gender, age and years of computer use on the occurrence of symptoms. Among other determinants, forearm support explained pain of wrist/hand, wrist support of elbow pain, and chair adjustment of arm pain. Association was also found between low back pain and chair adjustment and keyboard position. CONCLUSIONS: The findings revealed frequent occurrence of symptoms of pain in upper extremities and neck in office workers who use computers on a regular basis. Seating position could also contribute to the frequent occurrence of back pain in the examined population. PMID- 19999040 TI - [Vaccination against influenza in medical staff of Warsaw university hospitals and in students of Warsaw Medical University]. AB - BACKGROUND: According to CDC recommendations, medical staff should be vaccinated against influenza each year. The objective of this work was to establish the percentage of medical personnel and students vaccinated against influenza in two Warsaw university hospitals and Warsaw Medical University, to evaluate their level of awareness about the importance of vaccination and to identify reasons for not taking vaccines. MATERIAL AND METHODS: The anonymous questionnaire survey was conducted in the population of 166 physicians, 104 nurses and 254 students. RESULTS: Only 22.3% of physicians, 10.6% of nurses and 13.4% of students regularly take vaccines against influenza. Free of charge vaccination was offered to 19% of physicians, 15% of nurses and 6% of students. Awareness of the need for influenza vaccination and willingness to take free-of-charge vaccination were declared by 42% of nurses. Physicians justified their decisions not to undergo vaccination by no need to vaccinate, laziness and lack of time, nurses by no need to vaccinate and costs, and students by costs and laziness. CONCLUSIONS: The percentage of medical staff of Warsaw university hospitals who regularly take vaccines against influenza is low. In view of the reluctance to take influenza vaccines observed in the study population, there is an urgent need to intensify educational efforts to increase awareness of the importance to be vaccinated. PMID- 19999042 TI - [EPINet software for reporting occupational exposures to blood among medical personnel--rationale for its wide implementation in Poland]. AB - Although there is a lack of adequate surveillance data at the national level, the available results from some current studies provide examples that sharps injuries sustained by medical personnel create a significant problem in Poland. However, there has been relatively little attention focused on this issue. The paper describes Exposure Prevention Information Network (EPINet) developed in the United States in 1991 and adopted in nearly 50 other countries, including 13 European ones. The principles and advantages of EPINet are discussed as well as its effects in the context of preventing occupational exposures among health care workers. The possible barriers to the adoption of EPINet in Poland are explained and arguments supporting its implementation, taking account of local needs, are presented. PMID- 19999041 TI - [Occupational contact dermatitis in hairdressers]. AB - Numerous data from the medical literature show that working as a hairdresser is associated with the highest risk of occupational contact dermatitis. In Europe, hairdressers rank first of all occupation groups with the highest prevalence of occupational dermatitis. It is estimated that 10-20% and even 50% of hairdressers are affected with skin disorders. Skin problems occur soon after commencing hairdressing, in the first 2 years of work, sometimes during vocational training. The most common factors contributing to skin damage include water, shampoos, detergents, conditioners, hair dyes, bleaches, permanent wave solutions and components of gloves. Water and wet work are prime skin irritants - causal factors of irritant contact dermatitis while p-phenylenediamine, glyceryl monothioglycolate, ammonium persulfate and also fragrances, preservatives, disinfectants and metals are the most important allergens, which induce allergic contact dermatitis. PMID- 19999043 TI - [Risk of neoplastic diseases in conditions of exposure to radio- and microwave fields--epidemiologic investigations]. AB - The review presents current state of knowledge of cancer morbidity in persons exposed to radio- or microwave radiation in the environment or at working posts. The problem of cancer morbidity in persons exposed to microwaves was for many years limited to occupational medicine, but the introduction of mobile telephone system in the 1980s considerably increased the number of persons exposed to this radiation and raised concerns about cancer risks in the users of mobile phones. Cancer risks of microwaves are minor and doubtful, but there exist reliable epidemiologic investigations providing evidence that multiyear occupational exposure and intensive use of mobile phones for more than 10 years may cause the increased morbidity of certain types of neoplasms, mainly brain tumors, as well as hematopoietic and lymphatic malignancies. Neverthelesscurrent state of knowledge does not allow, according to the IARC classification, to classify radio and microwave fields as the carcinogenic factor. PMID- 19999044 TI - [Pictorial health warnings on tobacco products packages as a part of tobacco epidemic control]. AB - The use of tobacco products has been described by the World Health Organization as the risk factor responsible for six out of eight causes of death in the world. Informing about the harm of smoking may be presented in many ways like media campaigns, text warnings, or graphic warnings placed on tobacco products. The aim of this article is to describe the role of graphic warnings placed on tobacco products in light of tobacco control. In this work, the available current data, including researches and reports of WHO, have been used. Graphic warnings may be a very valuable source of knowledge about health consequences of smoking. They are also much more visible, draw attention much better compared with text warnings and more clearly communicate the threats of tobacco use. They also have a stronger impact, are better memorized and better motivate to quit smoking. Pictorial warnings are also approved by the society. Smokers themselves perceive them as more effective than text warnings. Moreover, this kind of labeling makes tobacco products less attractive. The introduction of legal regulations enforcing these types of warnings does not cost anybody but the tobacco companies. This kind of solution helps to gain public acceptance for other methods of tobacco control like totally free tobacco smoke places. PMID- 19999046 TI - [Poisonings with Amanita phalloides]. AB - Amanita phalloides is the most dangerous, poisonous mushroom species in our climatic conditions. It is the cause of 90-95% of all deaths due to mushroom poisoning, a-Amanitin, a polymerase RNA II inhibitor, is mainly responsible for the Amanita phalloides toxic property. Inhibition of polymerase RNA II functioning in a transcription process is connected with inhibition biosynthesis of structural and enzymatic proteins in cells. A lethal dose of a-amanitin is 0.1 mg/kg b.w. for humans. One of the medical problems in Amanita phalloides poisonings is a relatively prolonged latency period (8-24 h) from mushroom ingestion, at the same time the cytotropic action of absorbed toxins is revealed. In severe cases, multi organ failure, renal and hepatitis failure can occur. Deaths in a-amanitin poising cases follows between 6-16 days after intoxication. Mortality in this group of patients is still high and amounts to approximately 20 30% in adults and exceeds 50% in children. If mushroom poisoning occurs, it is best treated with pharmacological agents, extracorporeal methods for toxin removal and liver transplantation. Recent high expectations concerning liver albumin dialysis (based on MARS) should support liver regeneration and will also help with possible liver transplantation. In a medical community it is generally believed that every suspected Amanita phalloides poisoning should be referred to a specialized health center. PMID- 19999045 TI - [Psychological support for road accident participants: the program implementation outcome]. AB - Road accident belongs to one of the categories of traumatic events, and can cause posttraumatic stress disorder (PTSD). The most common psychological consequences of road traffic accidents are the emotional disorders, cognitive deficits (disorders concentration of attention and memory function), impaired social relationships, troubles with performing work duties and physical symp-toms of stress. The article discusses the program of psychological support given to the participants of road accidents, conducted in Wroclaw, its theoretical assumptions and the mode of its implementation. Basic theoretical assumptions of the program are inter alia based on the theories of H. Selye and R. Lazarus. The authors of this article also refer to the so-called therapeutic process of education designed by Everly and Rosenfeld, who recommend that the process of developing self-responsibility be used for therapeutic purposes. This requires clarifying the exact nature of the problem, and then looking for possible remedies. The program is open to all road accident participants (victims, perpetrators, witnesses) and their families. Classes are designed to combine theory (lectures) and practice (exercises). Anxiety and cognitive processes, as well as relaxation training, interpersonal training (eg, assertiveness) and kinesiology are the major areas of activities. Psychological support provided for road accident participants is of intervention--and preventive nature; intervention, as it relates to the consequences ofa specific stressogenic event, namely a road accident; preventive, as it serves the overriding purpose--the improvement of road safety. This article presents the main findings of the program, the results of the survey evaluation, and proposals to develop psychological operations aimed at the road accidents participants. PMID- 19999047 TI - [Health literacy as one of the contemporary public health challenges]. AB - One of the fundamental public health challenges in the 21st century should be the improvement of people's health literacy, namely the understanding of health messages. The acquired high level of health literacy means that one knows how and where information concerning health determinants can be found, is able to assess it critically and in favorable conditions even modify them, which seems to be of particular importance from the perspective of heath promotion, prevention or treatment of diseases. Therefore, for professionals in these fields, knowledge of ways how to improve health literacy, as well as awareness of related benefits and the consequences of its poor level, seems to be indispensable. Thus, the aim of this paper is to explain the term of "health literacy", its determinants and implications. PMID- 19999048 TI - More on continuous oral contraception. PMID- 19999049 TI - A rapidly spreading rash on a child. Strep infections, gastroenteritis precede symptoms. PMID- 19999050 TI - From exam table to council table. An election success story. PMID- 19999051 TI - Lower extremity amputations. Black men with diabetes overburdened. PMID- 19999053 TI - Patient information. Chronic kidney disease are you at risk? PMID- 19999052 TI - Mesotherapy for body sculpting. European staple of reshaping gaining ground here. PMID- 19999054 TI - Insulin replacement therapy. Minimizing complications and side effects. PMID- 19999055 TI - Type 2 diabetes in children and adolescents. Literature shows screening is overlooked. PMID- 19999056 TI - Pressed for time. Treating type 2 diabetes in the real world. PMID- 19999057 TI - What went wrong? A retrospective on Exubera. PMID- 19999058 TI - Communication breakdown. An overview of autonomic neuropathy. PMID- 19999059 TI - Postmenopausal osteoporosis. The role of intravenous bisphosphonates. PMID- 19999060 TI - Laceration evaluation and repair. The basics. PMID- 19999062 TI - Life in the slow lane. How to recharge your energy. PMID- 19999061 TI - Bipolar disorder. Recognition and treatment in primary care settings. PMID- 19999063 TI - Housecalls 101. When you see a need, fill it. PMID- 19999064 TI - Changing reimbursement policies. Billing at the physician rate isn't worth it in the long run. PMID- 19999065 TI - Submandibular lymph node in child. Treatment in the absence of other symptoms. PMID- 19999066 TI - Life span vs. healthy life expectancy. Where did men go wrong? PMID- 19999067 TI - Collagen stimulators. Beyond Botox, Juvederm, Restylane and Perlane. PMID- 19999068 TI - Patient information. Eggs: a good source of choline. Why your body needs this nutrient. PMID- 19999069 TI - After the cure. Follow-up care for cancer survivors. PMID- 19999070 TI - Bioidentical hormone therapy. An overview of benefits and prescribing guidance. PMID- 19999071 TI - Choosing the right infant formula. Protein and fatty acids are two of many considerations. PMID- 19999072 TI - Health need s of the older cancer patient. Constructing a comprehensive geriatric assessment. PMID- 19999073 TI - Prenatal yoga. Guidance for providers and patients. PMID- 19999074 TI - Doing right by everyone. Ethical dilemmas at work. PMID- 19999075 TI - The value of convenience. Retail health is where we want to be. PMID- 19999076 TI - You, too, can survive the recovery audit contractors. PMID- 19999077 TI - Hospitals in demo project prepare for permanent RACs. PMID- 19999078 TI - Put lessons learned from the RAC demo to work. PMID- 19999079 TI - Care coordinators work 24/7 to increase efficiency, decrease length of stay. PMID- 19999080 TI - Hospital's medication program helps uninsured. PMID- 19999081 TI - Organization key when the RAC requests arrive. PMID- 19999082 TI - Low-cost strategies help improve flow. PMID- 19999083 TI - What are the biggest risks involving ED handoffs? PMID- 19999084 TI - OSHA enforcing N95 respirators for HCWs treating H1N1 flu patients. PMID- 19999085 TI - [Experience of postoperative adjuvant chemotherapy of lung cancer at outpatient department]. AB - In late years the cancer adjuvant chemotherapy shifts from an inpatient care to an outpatient treatment. For operated lung cancer patients, outpatient chemotherapy center has been working since October 2005 in our hospital. Chemotherapy regimens were carboplatin (CBDCA) + paclitaxel (PTX), CBDCA + gemcitabine (GEM), docetaxel (DTX) + tegaful-gimeracil-oteracil potassium (S-1), and GEM + vinorel bine (VRE). CBDCA was chosen instead of cisplatin (CDDP) and non-platinum doublets are also used because of less toxicity and more time saving. Adjuvant chemotherapy has been performed for a total of 25 outpatients. Twenty-two out of 25 completed chemotherapy. Neutrophilopenia was the most common toxicity and grade 3 or 4 neutrophilopenia was seen in 6 patients. Adjuvant chemotherapy of outpatients can be completed safely by the choice of a safe regimen, supportive therapy for the toxicity, and cooperation with the community medicine organization. Our chemotherapy regimen are thought to be feasible for postoperative lung cancer outpatients. PMID- 19999086 TI - [Paraplegia associated with acute aortic dissection complicated with pulmonary embolism and re-dissection]. AB - Paraplegia associated with acute aortic dissection is one of the most serious complications. We experienced a case of Stanford type A acute aortic dissection with paraplegia and cardiac tamponade because the dissection was already thrombosed, conservative therapy was chosen. After drainage of pericardial effusion, a spinal drainage tube was inserted. Eleven days later, pulmonary embolism and re-dissection occurred, and an emergency operation was performed. Although the operation was successful, paraplegia did not improve. Even if type A acute aortic dissection is complicated with paraplegia, early surgical repair of aortic dissection should be considered, when paraplegia does not improve rapidly in spite of treatment. PMID- 19999087 TI - [Emphysema evaluation of the remaining lung post lobectomy]. AB - The aim of this study was to confirm that the emphysematous changes had increased in the remaining lung on the operation side after lobectomy. Fourteen patients underwent quantitative analysis by computed tomography (CT) and respiratory function testing before and after the operation for upper or lower lobectomy of the lung between March 2005 and May 2007. The degree of emphysema was calculated by dividing the volume of the emphysematous region (CT values: -1,024 to -950 HU) by the volume of the entire lung (CT values: -1,024 to -600 HU) using a 1 mm thickness high resolution CT slice. Comparison by paired t-test showed significant differences between the emphysema rates pre and post operation in the operation side lung (15.3 +/- 7.9% and 21.7 +/- 10.0%, p = 0.02), but there were no significant differences in the contralateral lung (15.8 +/- 7.5% and 17.7 +/- 8.4%, p = 0.25). On the other hand, there was no significant change in the percent forced expiratory volume in one second (FEV1.0%) between pre and post operation (74.4 +/- 10.5% and 75.5 +/- 7.5%, p = 0.60). We consider that it is necessary to note that the emphysematous changes increased in the remaining lung on the operation side after lobectomy even though the FEV1.0% did not decrease at present. PMID- 19999088 TI - [Aortic root replacement for aortic dissection and aortic regurgitation due to aortitis syndrome]. AB - Aortic aneurysms and aortic regurgitation (AR) with aortitis syndrome are occasionally reported in young women. We report a case of aortic dissection with severe AR in an 8-year-old girl. The patient underwent aortic root replacement with a composite graft. Pathological report revealed aortitis syndrome and steroid therapy was continued to suppress further inflammatory vascular reaction. PMID- 19999089 TI - [Endarterectomy and aortic valve replacement in a patient with severely calcified ascending aortic root]. AB - A severely calcified, so-called, "porcelain" aorta may complicate aortic valve replacement (AVR). We report a case of successful AVR with an extensively calcified aortic root, involving the right coronary ostium. A 73-year-old female presented with a stenotic aortic valve with the pressure gradient of 60 mmHg. Computed tomography demonstrated extensive calcification of the ascending aortic root. Coronary angiogram showed 90% right coronary artery ostial stenosis. We endarterectomized the ascending aorta, the Valsalva sinus and the right coronary ostium, which enabled AVR with a 19 mm CarboMedics valve. These procedures might be a favorable option for the treatment of porcelain aorta and calcified aortic valve. PMID- 19999090 TI - [Intraoperative right-to-left shunting induced by supine position in simultaneous lung resection with repair of atrial septal defect]. AB - This report describes the case of a 49-year-old woman, who was suffered severe hypoxemia attributable to right-to-left shunting through an atrial septal defect (ASD) during the combined surgery for lung cancer and ASD in supine position. Right-to-left shunting has been reported to occur after lung resection but not during it. According to our continuous measurement of pulmonary arterial pressure and oxygen saturation, changes in hemodynamics during lobectomy in supine position was supposed to differ from that in lateral position, which may contribute to right-to-left shunting. A combined lung resection with heart surgery was performed safely and resulted in preventing postoperative complications induced by ASD. PMID- 19999091 TI - [Large saphenous vein graft aneurysm after coronary artery bypass grafting]. AB - A 64-year-old man underwent coronary artery bypass grafting (CABG) in 1994. Eleven years later, coronary angiography (CAG) showed a saphenous vein graft (SVG) aneurysm. In April 2008, the CAG demonstrated enlargement of the aneurysm. Enhanced computed tomography showed a large aneurysm measuring 53 x 44 mm in diameter of the SVG-right coronary artery (RCA) [#3] anastomosis. Off-pump coronary surgery to RCA and aneurysmectomy was planned through a median epigastric incision. However re-grafting was difficult because of severe adhesion and calcification of the coronary artery. We confirmed sufficient back flow from the distal coronary artery without hemodynamic and electrocardiographic changes under SVG clamp. Aneurysmectomy was then performed without coronary bypass. The patient was discharged 9 days after surgery and has been well for 8 months in outpatient clinic. PMID- 19999092 TI - [Coronary artery aneurysm with various clinical course]. AB - Case 1: A 77-year-old woman had effort angina pectoris. Coronary angiography (CAG) revealed a coronary artery aneurysm on the left descending artery. Coronary artery bypass grafting (CABG) and patch angioplasty for the aneurysm were performed. Case 2 : A 69-year-old woman had effort dyspnea CAG showed dilation of the left main trunk and beaded aneurysms (maximum 6 cm in diameter) behind the ascending aorta with a fistula to the right atrium. We closed the fistula and performed CABG to the circumflex branch. Case 3 : A 78-year-old woman had had general fatigue for 2 weeks. Previous CAG had revealed coronary artery aneurysms and current chest computered tomography revealed pericardial effusion. She was, therefore, diagnosed with the rupture of the coronary artery aneurysm. We closed the coronary artery aneurysm and performed CABG. Case 4: A 55-year-old man had been diagnosed with acute myocardial infarction and had undergone percutaneous coronary intervention 3 years before. CAG revealed a coronary artery aneurysm on the right coronary artery. We resected the aneurysm and interposed with saphenous vein graft. Although coronary artery aneurysm often has no symptoms, in the cases of angina, myocardial infarction, rupture or large aneurysm more than 3 times larger than the normal diameter, surgical repair should be considered. PMID- 19999093 TI - [Lung metastasis of thymic carcinoma twelve years after complete resection of primary disease]. AB - We report a 67-year-old woman who developed lung metastasis 12 years after resection of the mediastinal tumor which was diagnosed as B3-thymoma. Combined resection of bilateral lungs and left innominate vein was performed due to tumorimvasion. 12 years later, she was diagnosed as having primary lung adenocarcinoma with bronchoscopic examination and cytology. Postoperative histological and immunohistochemical examination showed that the lung nodule was a lymphoepithelioma-like carcinoma of the thymus with strong immunoreactivity for CD5 and bcl-2. Reexamination by immunohistochemical staining for CD5 and bcl-2 revealed that the disease diagnosed as thymoma 12 years ago was also thymic carcinoma Accurate differential diagnosis between B3-thymoma and thymic carcinoma is essential for choosing appropriate postoperative treatment. PMID- 19999094 TI - [Thymolipoma associated with myasthenia gravis]. AB - A 42-year-old woman was admitted to our hospital complaining of left ptosis, diplopia, and muscle weakness. A diagnosis of myasthenia gravis was made. Chest roentgenograms, computed tomography (CT), and magnetic resonance imaging (MRI) showed a large anterior mediastial mass and suggested thymolipoma. Extended thymectomy was performed via a median sternotomy. Histopathological examination revealed that the tumor consisted of mature adipose tissue and weighed 1,500 grams, in which thymic tissues with Hassall' s corpuscles but without a germinal center were contained. The histological appearance was compatible with a typical thymolipoma. This is the 24th reported case of thymolipoma associated with myasthenia gravis. PMID- 19999095 TI - [Bronchogenic cyst with a high computed tomography number]. AB - A 55-year-old man was diagnosed as having colon cancer. During the preoperative survey for metastasis, chest computed tomography (CT) revealed a well-defined tumor (33 mm) with a high CT value in the left hilum. We performed thoracoscopic resection for diagnosis and treatment. The contents of the lesion were yellowish, viscid, and creamy. Pathological examination revealed that the tumor was a bronchogenic cyst. This case is reported with a review of the literature. PMID- 19999096 TI - [Thoracic aortic rupture resulting from methicillin-resistant Staphylococcus aureus infection: report of a case]. AB - We report a successful surgical treatment of an aortic arch rupture caused by methicillin-resistant Staphylococcus aureus (MRSA) infection in a 72 years old man who had back pain and hemoptysis as symptoms. Computed tomography showed enlargement of the aortic arch within just 4 days. We diagnosed the state of impending rupture of inflammatory aneurysm and performed emergent operation. The upper lobe of the left lung adhered firmly to the site of rupture and sealed as the barrier to prevent bleeding. The aorta was replaced with a dacron graft in situ after entire resection of infected tissue and sufficient irrigation of the pleural cavity. The patient was treated with vancomycin and minocyclin for MRSA infection. The postoperative course of the patient was uneventful. This suggests that an early operation and a proper use of antibiotics lead to a successful clinical result in patient with infectious aortic rupture. PMID- 19999097 TI - [Ascending aortic aneurysm and aortic valve regurgitation long after operative repair of coarctation of the aorta: report of a case]. AB - We report a case of surgical treatment for ascending aortic aneurysm and aortic valve regurgitation (AR) 24 years after operative repair of coarctation of the aorta (CoA). The patient was a 32-year-old man who had undergone operative repair of CoA and patent ductus arteriosus (PDA) ligation when 8 years old, and was followed since then. However, since 14 years after the operation, dilation of his ascending aorta and AR was observed. Then the AR deteriorated and the ascending aorta dilated, and at 24 years after operation he had symptoms of heart failure. So we performed ascending aorta replacement and aortic root replacement (reimplantation). Despite primary success of the operative repair of CoA, however. 9% of patients develop aortic aneurysms long after the operation. Therefore, long-term follow-up is needed after repair of coarctation of the aorta. PMID- 19999098 TI - [Constrictive pericarditis after minimally invasive coronary artery bypass grafting: report of a case]. AB - Constrictive pericarditis (CP) after off-pump coronary bypass surgery, especially after minimally invasive direct coronary artery bypass (MIDCAB), had rarely been reported. We presened a surgically treated case of CP after MIDCAB via left anterior small thoracotomy. A 57-year-old man underwent MIDCAB with placement of an internal mammary artery to the left anterior descending coronary artery uneventfully. Four years after the operation, he began to experience exertional dyspnea. Computed tomography of the chest showed pericardial thickening. Cardiac catheterization revealed elevation and equalization of the pressures in the 4 chambers, as well as low cardiac output. Pericardiectomy using cardiopulmonary bypass through a median sternotomy was performed successfully without injury to the bypass graft. Postoperative hemodynamic measurements were improved. The patient has resumed normal activity and remained free from heart failure for over 5 years. PMID- 19999100 TI - [Primary clear cell adenocarcinoma of the lung with endobronchial polypoid growth: report of a case]. AB - Clear cell adenocarcinoma with endobronchial polypoid growth of the lung is extremely rare. A 65-year-old male with hemosputum was found to have an abnormal shadow in the hilum of the left lung. Computed tomography of the chest revealed that a heterogeneous mass occupied the lumen extending outside the upper lobe bronchus of the left lung. By biopsy, the tumor was determined to be adenocarcinoma. The patient underwent left pneumonectomy with mediastinal lymph node dissection. Macroscopically, the tumor showed a polypoid growth along with the bronchial tree. Microscopically, most of the tumor was composed of large clear cells with partial glandular formation, indicating the tumor to be adenocarcinoma Lymph node metastasis was seen in #5 and #12u. The lung cancer was diagnosed as clear cell adenocarcinoma with endobronchial polypoid growth. PMID- 19999099 TI - [Cardiac surgery in an octogenarian with idiopathic thrombocytopenic purpura: report of a case]. AB - An 81-year-old woman was referred to our hospital for surgical treatment for mitral valve regurgitation, tricuspid valve regurgitation and atrial fibrillatory bradycardia. The platelet count on admission was 4.9 x 10(4)/microl. and the results of other studies were compatible with idiopathic thrombocytopenic purpura. Although we performed high-dose transvenous immunoglobulin infusion (400 mg/kg/day) for 5 consecutive days, the platelet count showed no remarkable change. Because of progression of heart failure, we underwent cardiac operation under thrombocytopenic condition. Intra and post-operative platelet transfusion might contribute to postoperative course uneventful without bleeding tendency. In this case, high-dose immunoglobulin therapy was not effective. However the operative course was satisfactory with adequate surgical hemostasis and platelet transfusion. PMID- 19999101 TI - [Peripheral intrapulmonary lipoma: report of a case]. AB - A 62-year-old woman who had undergone a left mastectomy for a breast cancer consulted us for an abnormal chest shadow. Chest computed tomography showed a well-defined nodule of 1 cm diameter periphery in lower lobe of the right lung. The differential diagnosis included benign lung tumors, such as intrapulmonary lymph nodes, granuloma etc. However, because of her past history we needed to consider metastasis. To make diagnosis, a wedge resection of the pulmonary nodule was performed. The tumor was diagnosed as a lipoma. No malignant cells were seen. Although peripheral intrapulmonary lipoma is very rare, it should be kept in mind in the differential diagnosis of an intrapulmonary nodule. PMID- 19999103 TI - [Molecular structure of Helicobacter pylori]. PMID- 19999102 TI - [Mediastinal germ cell tumor with somatic-type malignancy successfully treated with chemotherapy followed by surgical resection combined with lobectomy: report of a case]. AB - A 28-year-old male visited our hospital with complaint of chest pain. Clinical examination revealed a huge mediastinal mass which was diagnosed as non seminomatous germ cell tumor. The patient underwent 5 cycles of chemotherapy (bleomycin, etoposide, and cisplatin) followed by resection of the tumor combined with left upper lobectomy. Final pathological diagnosis was germ cell tumor with somatic-type malignancy. While the prognosis of mediastinal germ cell tumor with somatic-type malignancy is known to be extremely poor, multimodality at an early stage is the key to successful treatment. PMID- 19999104 TI - [Guidelines in the management of H. pylori infection in Japan--2009 version]. AB - The results of a large-scale multicenter study conducted by Japanese H. pylori investigators were published in the Lancet in 2008. Patients after endoscopic treatment for early gastric cancer were randomized to eradication and non eradication groups for investigation of recurrence after 3 years, demonstrating that recurrence was markedly inhibited in the eradication group. The question asked all over the world about whether H. pylori eradication can inhibit the onset of gastric cancer could be definitely answered by these results. For preparation of these guidelines, the addition of H. pylori infection as an indication for eradication was discussed, and it was unanimously accepted as an additional indication. PMID- 19999105 TI - [Epidemiology of H. pylori infection and exploration of its infection route]. AB - To build up a preventive strategy for H. pylori related diseases including gastric cancer, infection ages and routes are important. Recently, several studies have explored them. In advanced countries, most infections occur under five years of age. Mother to child in some studies and sibling to sibling in others were dominant infection routes. As infection ages and routes depend on countries, studies in Japan are indispensable. Infection in kindergartens, nursery and elementary schools as well as in families should be explored. Based on the findings, a strategy preventing infection to children should be build up to prevention H. pylori related diseases. PMID- 19999106 TI - [Helicobacter pylori infection in childhood]. AB - Helicobacter pylori (H. pylori) infection is mainly acquired in the first 2 or 3 years and the risk of infection declines rapidly after 5 years of age. In developing countries, acquisition age of the infection is probably lower than in developed countries. In Japan, main transmission route is intrafamilial and mother to children infection is most important. But in developing countries, some reports suggest that extrafamilial infection is more important. The famous paper revealed that H. pylori can be cultivated from vomitus, saliva and cathartic stools and the possibility of source of H. pylori infection. Bed sharing, large number of family members, delayed weaning from a feeding bottle, regurgitated gastric juice in the mother's mouth are reported as risk factors of the infection. PMID- 19999107 TI - [The role of CagA in H. pylori infection]. AB - Helicobacter pylori (H. pylori) chronically colonizes human gastric epithelium and induces various diseases. But the mechanism of carcinogenesis in H. pylori infection remains to be assessed. We described that after attachment of H. pylori to gastric epithelial cells, CagA is injected directly from the bacteria into the cells and undergoes tyrosine phosphorylation. Tyrosine phosphorylated CagA can bind to SHP-2. Deregulation of SHP -2 by CagA may induce abnormal proliferation and movement of gastric epithelial cells. There are two patterns of CagA motifs between East Asian strains and Western strains. East Asian-type CagA confers stronger SHP-2 binding and transforming activities than Western-type CagA. We assessed the association between CagA diversity and clinical outcome in Asian countries, where mortalities from gastric cancer is different. As results, H. pylori infection with East Asian-type CagA was associated with gastric atrophy and cancer. Therefore, persistent active inflammation induced by the East Asian CagA-positive strain may play a role in the pathogenesis of disease. PMID- 19999108 TI - [Helicobacter pylori virulence factors except CagA]. AB - Study for Helicobacter pylori pathogenesis has been already performed widely all over the world for about 20 years. This review summarized recent studies about virulence factors except CagA/cag PAI system. Bacterial adhesion to gastric mucosa is critical for the persistent infection and BabA, SabA, OipA, AlpA or AlpB is involved. VacA (vacuolating cytotoxin) causes not only vacuolation of gastric epithelial cells but also stimulates apoptosis via a mitochondria dependent pathway and is a potent immunomodulatory toxins. Signaling pathway via toll-like receptors(TLR) is also thought to be important especially in myeloid cells even though bioactivity of H. pylori LPS is reported to be weak compared with other bacteria such as E. coli. PMID- 19999109 TI - [Role of host genetic factors in gastric carcinogenesis]. AB - Helicobacter pylori infection is the most common chronic bacterial infection worldwide and the most important acquired risk factor for gastric cancer. Bacterial, environmental and host genetic factors combine to define the degree of gastric damage. The most relevant and consistent genetic factors are in the interleukin-1B and tumor necrosis factor-A gene clusters. These cytokines play a key role in the gastric carcinogenesis and their roles have been confirmed in mouse models. More genetic factors have also been uncovered recently and certain cytokine and innate immune response gene polymorphisms appear to influence risk of gastric cancer and its precursor conditions. In the near future, we will be able to define a more comprehensive genetic profile that would identify high risk groups of developing gastric cancer. PMID- 19999110 TI - [A novel mechanism for H. pylori-induced cell damage--induction of gene mutation by AID]. AB - H. pylori infection causes gastric cancer development through gastritis. However, the precise mechanism for H. pylori-induced carcinogenesis remains unclear. On the other hand, accumulation of gene mutations is a hallmark of cancers, but mechanism of endogenous induction of mutation also remains unknown. Here, we found that H. pylori infection triggers aberrant expression of activation-induced cytidine deaminase (AID), a molecule that can induce DNA somatic hypermutation of immunoglobulin gene in B cells under physiological condition, in the gastric mucosa via NFkappaB-dependent mechanism. Induction of AID then enhances various gene mutations including p53. This mechanism appears to play important roles in gastric carcinogenesis by H. pylori infection. PMID- 19999111 TI - [Latest advances in non-pylori Helicobacter species]. AB - The genus Helicobacter presently comprises 32 validly named species. Helicobacter equorum, Helicobacter baculiformis and Helicobacter suis were recognized as new species in 2007 and 2008. Non-pylori Helicobacter species are occasionally isolated from human clinical specimens, though their natural hosts are mammals other than human and birds. It remains unclear whether they are associated with human diseases. 'Helicobacter heilmannii' causes human chronic gastritis. Several reports suggest that Helicobacter hepaticus and Helicobacter bilis are associated with human hepatic and biliary disorders, respectively. Non-pylori Helicobacter species are occasionally isolated from clinical specimens of immunocompromised hosts. These cases suggest that they are important opportunistic pathogens. PMID- 19999112 TI - [Diagnostic methods for H. pylori infection]. AB - The diagnosis of H. pylori infection is based on methods requiring gastric biopsies performed during endoscopy (rapid urease test, histology, culture) or on non-invasive methods (antibody, 13C urea breath test, stool antigen test). The accuracy of each method varies and each method has various merits and demerits, so it's important to select proper method according to the situation and purpose of the diagnosis of the infection. PMID- 19999113 TI - [Diagnosis of H. pylori infection in children]. AB - There are two types of diagnostic tests used to detect H. pylori infection: non invasive and invasive in children. Non-invasive tests include the urea breath test, stool antigen tests, and blood or urine serologic assays which detect the presence or absence of infection. The urea breath test and stool antigen test have adequate sensitivity and specificity in children and recommended to detect active H. pylori infection before and after eradication therapy. Invasive tests include the performance of upper gastrointestinal endoscopy with gastric biopsy. The histopathology can determine both presence or absence of infection and the extent and severity of mucosal injury (i.e., disease). Culture of gastric tissue has 100% specificity and the antimicrobial susceptibility test is available. Non invasive tests are developed for use in children, the use of upper gastrointestinal endoscopy with gastric biopsy remains the diagnostic strategy of choice in symptomatic children. PMID- 19999114 TI - [The present status and problems of Helicobacter pylori first-line eradication therapy]. AB - The widespread of eradication therapy for H. pylori has lead to an increase in antibiotic resistant strains. The combination of a proton pump inhibitor, amoxycillin and clarithromycin (PPI/AC) has been one of the most popular regimens and has shown satisfactory results. Clarithromycin (CAM)-resistance of H. pylori is a clinical problem because it reduces the efficacy of PPI/AC regimen. We investigated the rate of acquisition of H. pylori resistance against CAM before triple therapy. The rate of CAM-resistant strains was 0% in 1985, 14.6% in 1996 99, 21.2% in 2001-04, and 24.1% in 2005-08, respectively. Tokyo H. pylori Study Group consisting of eleven institutions assessed the eradication rate of PPI/AC regimen from 2001 to 2008 in Tokyo metropolitan area. Intension-to-treat based eradication rate for PPI/AC regimen was decreased gradually from 70% to 60% level. The reduction in eradication rate of H. pylori consisting of PPI/AC was recognized. Furthermore, when the increase in CAM-resistant strains results in a decrease in eradication rate for H. pylori, new alternative first-line treatment combination should be considered in Japan. PMID- 19999115 TI - [Second-line therapy for patients with Helicobacter pylori eradication failure]. AB - Meta-analyses showed that quadruple therapy, ranitidine bismuth-based triple therapy and levofloxacin-based triple therapy were the most effective for the second-line H. pylori eradication therapy. In Japan, many studies showed that triple therapy with proton pump inhibitor (PPI), amoxicillin (AMPC) and metronidazole was effective after eradication failure using triple therapy with PPI, AMPC and clarithromycin. The intention-to-treat eradication rates were 81 97%, and the incidence rates of adverse events were 8-33%. PMID- 19999116 TI - [Present status and future prospect of 3rd rescue regimen for H. pylori infection]. AB - The standard strategy for H. pylori infection in Japan consists of the triple PPI/AMPC/CAM therapy as the first line therapy and the triple PPI/AMPC/MNZ therapy as the second line therapy. 2 or 3% of patients cannot achieve eradication of H. pylori with the above two regimens. Most of them are infected with CAM resistant strains of H. pylori and have the rapid metabolizer genotype of CYP2C19. There are two major third line therapies. One is the dual therapy with high doses (qid dosing) of PPI and AMPC. The other is fluoroquinolone-based therapy. Especially, sitafloxacin, one of the fluoroquinolones, has the lower MIC for H. pylori in comparison with other fluoroquinolones, such as LVFX. Both therapies are expected to achieve the high eradication rate as the 3rd line therapy. PMID- 19999117 TI - [Helicobacter pylori eradication therapy in children]. AB - Proton pump inhibitor-based triple therapies are performed worldwide for Helicobacter pylori-associated diseases in childhood. As the first-line therapy, the Japanese Pediatric Guideline in H. pylori Infection has recommended a triple regimen with amoxicillin and clarithromycin, because the Japanese national health care insurance system has permitted only this regimen. However, the primary resistance to clarithromycin in Japan is demonstrated in around 30% of the strains, leaving an urgent issue for the physicians. A triple regimen with amoxicillin and metronidazole is acceptable as the second-line therapy. In children with chronic H. pylori gastritis, eradication should be considered if they have gastric atrophy or a family history of gastric cancer. In children, however, a 'test and treat' strategy to prevent gastric cancer is not recommended. PMID- 19999118 TI - [Gastritis]. AB - Eradication of Helicobacter pylori (H. pylori) has suggested not only the decrease in the incidence of peptic ulcer diseases and gastric cancer, and but also the improvement of dyspeptic symptoms. Dyspepsia is a common condition in the world, and dyspepsia-related medical cost is high due to consultations, investigations, and drug consumption. Although the role of H. pylori in functional dyspepsia is still unknown, recent studies including meta-analysis have indicated that H. pylori eradication improves dyspeptic symptoms. In addition, the reduction of medical cost by H. pylori eradication has been discussed in the more recent studies. PMID- 19999119 TI - [Helicobacter pylori eradication therapy for peptic ulcer disease]. AB - Meta-analyses showed that Helicobacter pylori eradication therapy was effective for healing and prevention of recurrence of peptic ulcers in H. pylori-positive patients and that treatment of H. pylori infection was more effective than antisecretory non-eradicating therapy (with or without long-term maintenance antisecretory therapy) in preventing recurrent bleeding from peptic ulcer. H. pylori eradication therapy is effective for prevention of development of NSAID ulcers in NSAID-naive patients. For prevention of recurrence of NSAID-ulcers, therapy with proton pump inhibitor is effective. PMID- 19999120 TI - [Gastric MALT lymphoma: recent topics regarding treatment and genetic aberrations]. AB - The recent topics in the treatment and genetic aberrations of gastric MALT lymphoma were reviewed. The chromosomal translocation t(11;18) (q21;q21)/API2 MALT1 has been frequently detected not only in gastric MALT lymphoma cases, but also in gastric diffuse large B-cell lymphoma (DLBCL) cases in two studies. Conversely, IGH-involved translocations, including t(3;14) (p14;q32)/FOXP1-IGH, are rarely seen in gastric MALT lymphoma. Several large-scaled clinical series have revealed that Helicobacter pylori eradication therapy is the optimal first line treatment for patients with gastric MALT lymphoma, which leads to a favorable long-term outcome. A number of modalities and/or regimens, including chemotherapy, immunotherapy, and radioimmuotherapy, have been developed as the second-line treatments for patients not responding to eradication therapy. PMID- 19999121 TI - [Gastric cancer]. AB - From many findings, it has been established that persistent infection of Helicobacter pylori (H. pylori) causes chronic active gastritis and subsequently causes the gastric mucosa of the high risk for gastric cancer development. On the other hand, recent Japanese-study results have shown the possibility of gastric cancer prevention by H. pylor eradication. Moreover the development of gastric cancer in uninfected subjects is very rare; therefore, prevention of gastric cancer by H. pylori eradication becomes a topic in Japan. To get rid of gastric cancer from Japan, the risk of gastric cancer should be determined by presence of H. pylori infection in a young fellow, on the other hand, the risk by the examination that combined serum PG method with serum antibody method in subjects after middle aged. It is now expected that eradication treatment should be performed for these high-risk subjects. PMID- 19999122 TI - [Idiopathic thrombocytopenic purpura]. AB - Several recent studies have suggested the eradication of Helicobacter pylori (H. pylori) can increase the platelet count in patients with chronic idiopathic thrombocytopenic purpura (ITP). A systematic review of the literature published in English shows that overall platelet responses were observed in over 50% of the patients after successful eradication. The response rate tended to be higher in countries with a high prevalence ratio of H. pylori infection in background population, in patients with less severe degree of thrombocytopenia and in those with shorter disease duration. The detection and eradication of H. pylori should be considered as the first line of treatment in all Japanese patients with chronic ITP. PMID- 19999123 TI - [Iron-deficiency anemia in children]. AB - Recent studies including case reports, case series, observational epidemiologic studies and intervention trials have suggested an association of Helicobacter pylori (H. pylori) and iron-deficiency anemia (IDA). Resolution of IDA was reported to be achieved among the patients who had a suboptimal response to iron therapy despite adequate iron supplementation or who had frequent relapses after stopping iron supplementation in children, especially in puberty. The biologic mechanism by which H. pylori induces the iron stores is not fully understood, but we recently reported that strains of H. pylori derived from pediatric patients with IDA showed enhanced Fe ion uptake and Fe ion-dependent rapid growth compared with those from patients with non-IDA. PMID- 19999124 TI - [Helicobacter pylori infection and allergic diseases]. AB - The prevalence of airway allergic diseases has increased in developed countries including Japan. There is a much published potential link between allergy and childhood infection. In 1989, Strachan proposed "hygiene hypothesis" that infections and living in an unhygienic environment might prime the immune system and thus protect against the development of allergic diseases. H. pylori would suppress allergic immune-mediated inflammation through T helper type 1 (Th1)/Th2 paradigm, which is suppression of Th2 responses by stimulation of Th1 immunity. In contrast, H. pylori infection has been associated with extra-digestive pathologies including chronic idiopathic urticaria. Although systematic review demonstrated that eradication was associated with remission of urticaria, the association is still controversial and the pathogenic mechanisms have never been confirmed. PMID- 19999125 TI - [Gastroesophageal reflux disease (GERD). Barrett esophagus]. AB - Gastroesophageal reflux disease (GERD) is defined as a troublesome condition caused by reflux of gastric contents into the esophagus. Reflux esophagitis and Barrett esophagus are included into complication of GERD. The relationship between H. pylori infection and GERD has been evaluated in many studies. H. pylori infection rate is generally low in patients with reflux esophagitis or Barrett esophagus. The increase in the incidence of reflux esophagitis after successful eradication of H. pylori has been alleviated by several reports from Western countries. However, temporary appearance or aggravation of reflux esophagitis after H. pylori eradication has been reported in Japan because of acid secretion increases. After long-term follow-up, most of them remain in mild severity of reflux esophagitis, and symptoms may not become more severe. The increase in symptoms of GERD rarely occurs after eradication of H. pylori. Therefore, the presence of reflux esophagitis is not an impediment to performing eradication therapy. PMID- 19999126 TI - [Lifestyle-related diseases and H. pylori]. AB - H. pylori infection elicits a chronic cellular inflammatory response not only in the gastric mucosa, but also in the extra-digestive organs. Indeed, H. pylori infection has been epidemiologically linked to the extra-digestive conditions and diseases. In this article, we review recent progress in our understanding about the relationship between H. pylori infection and lifestyle-related diseases, such as atherosclerosis, diabetes mellitus, obesity, hyperlipidemia, hypertension, and osteoporosis. Although the relationships between all of these diseases and H. pylori infection are still controversial, H. pylori infection may increase the risk for cardiovascular and cerebrovascular diseases, especially when CagA+ cytotoxic strains of H. pylori are present. PMID- 19999127 TI - [Acquisition of the drug resistance and Helicobacter pylori gene mutation]. AB - The currently recommended regimen in Japan is a proton pump inhibitor-based triple therapy with two of the antibiotics between clarithromycin, amoxicillin and metronidazole. However, recent years have witnessed a decrease in the rate of eradication due to antimicrobial resistance. Resistance mechanisms of H. pylori are explained by the occurrence of mutations in genomic genes, which may correspond to the 23S rRNA gene in clarithromycin, the pbp1A in amoxicillin, and the rdxA in metronidazole, gyrA in levofloxacin. The resistance of H. pylori strains to clarithromycin is currently estimated at about 30% in Japan, while the resistant rates to metoronidazole and amoxicillin are quite low. The resistant rates of each antibiotics may increase in future, so we need to observe those changes. PMID- 19999128 TI - [Therapeutic trends of diseases of the upper gastrointestinal tract in patients with negative H. pylori status]. AB - Many diseases of the upper gastrointestinal tract developed in patients with Helicobacter pylori (H. pylori) infection, thus the conditions unrelated to H. pylori are rare. Here, we described the therapeutic trends of diseases in H. pylori negative individuals. Proton pump inhibitor (PPI) is superior to H2 receptor antagonist in both gastroesohageal reflux disease (GERD) including reflux esophagitis and non-erosive reflux disease (NERD) whereas therapeutic gains of PPI treatment for NERD patients are lower than those reported in GERD because of heterogeneity of NERD pathophysiology. Endoscopic therapy for PPI refractory GERD patients remains to be established because there are few studies concerning the effectiveness or safety of the procedures. Main cause of H. pylori negative ulcer diseases is the use of non-steroidal anti-inflammatory drugs (NSAIDs). PPI therapy is effective for both the prevention and treatment of NSAIDs-induced peptic ulcer. Considerations that should be entertained as causes of intractable peptic ulcers include Zollinger-Ellison syndrome or Crohn's disease. Gastric cancer and carcinoid tumor should be treated with endoscopic or surgical resection regardless of H. pylori infection. As for the treatment for gastric H. pylori-negative MALT lymphoma, radiation therapy (RT) should be selected first, and next chemotherapy will be given to the patients who failed to RT. PMID- 19999129 TI - [Recent proceedings in epigenetics research of lung cancer]. AB - Epigenetics is a mechanism of reversible and heritable regulation of gene transcription in contrast to genetic or DNA-coding information. In higher vertebrae, epigenetics consist of methylation of cytosine and histone modification. Mainly as detection is easier, aberrant DNA methylation was studied since the middle of '80s and recently convenient kit makes easy for study of histone modification. DNA methylation and histone modification meet in the end of '90s and regulation of transcription has being revealed. In this review, a history of epigenetics is commented briefly and recent proceedings of epigenetic change in lung cancer including micro RNA inactivation by DNA methylation and multigenetic analyses in non-small cell lung cancer is discussed. PMID- 19999130 TI - [Clinical experience with transurethral resection of bladder tumor (TUR-Bt) guided by photodynamic diagnosis (PDD)]. AB - PURPOSE: To report our clinical experience regarding transurethral resection of bladder tumor (TUR-Bt) guided by photodynamic diagnosis (PDD) with intravesical instillations of 5-aminolevulinic acid (ALA) and to assess the usefulness of the therapeutic method. MATERIALS AND METHODS: TUR-Bt guided by PDD was performed in 57 patients of which 47 were men and 10 women with a median age of 74.3 years (range 45-90), 36 were primary cases and 21 were recurrent cases with non-muscle invasive bladder cancer. Two to two and half hours prior to endoscopy 1.5 g ALA dissolved in 50 ml of 8.4% sodium hydrogen carbonate (NaHCO3) solution was instilled intravesically. For fluorescence excitation a blue light source (D LIGHT System, Karl Storz Endoscopy Japan K.K.) was used. The tumorous lesions under white light guidance and the lesion with fluorescent excitation under blue (fluorescence) light guidance were taken by cold cup as a biopsy and also resected sequentially. To evaluate the accuracy of PDD, the levels in images of the ALA-induced fluorescence were compared with the pathological results. To evaluate the availability of TUR-Bt guided by PDD, survival Analysis regarding vesical recurrence was retrospectively examined compared to the cases underwent conventional TUR-Bt under white light guidance. Moreover, in these cases, multivariate analysis using Cox proportional-hazards model was performed to detect the clinico-pathological factor independently contribute to improving prognosis. (Results) In the 301 specimens obtained from 57 patients, the sensitivity and specificity of PDD were 92.5% and 60.1%, whereas the sensitivity and specificity of conventional endoscopic examination under white light guidance were 81.6% and 79.5%, respectively. Median follow-up period was 19.1 (range 8.6 49.9) months in 57 patients underwent TUR-Bt guided by PDD. Eight of 57 patients recurred and recurrence-free survival rate was 88.2 +/- 0.1% (at 12 months) and 76.2 +/- 0.1% (24-48 months). Median follow-up period was 49.9 (5.0-145.0) months in 149 patients underwent conventional TUR-Bt. Ninety-nine of 149 patients recurred and recurrence-free survival rate was 60.3 +/- 0.0% (12 months) and 31.6 +/- 0.0% (24-48 months). There was statistical significance in recurrence-free survival rate between these 2 therapeutic groups (p < 0.001). Moreover, multivariate analysis revealed the independent factor contribute to improving prognosis was only TUR-Bt guided by PDD (hazard ratio 0.279, p = 0.001). CONCLUSION: It was suggested that TUR-Bt guided by PDD might reduce the risk of vesical recurrence in the early stage after operation of non-muscle invasive bladder cancer. PMID- 19999131 TI - [Treatment results of salvage radiotherapy for biochemical recurrence after radical prostatectomy]. AB - PURPOSE: In this retrospective study we reported the results of salvage external beam radiotherapy for patients with biochemical recurrence after radical prostatectomy. MATERIALS AND METHODS: A total of 28 patients with biochemical recurrence after radical prostatectomy underwent salvage radiotherapy with (n=16) or without (n=12) hormonal therapy. Median radiation dose was 60 Gy. Biochemical recurrence after radiotherapy was defined as a single prostate-specific antigen (PSA) of at least 0.1 ng/ml. Potential risk factors were evaluated for significant associations with biochemical recurrence. RESULTS: The median follow up period after salvage radiotherapy was 42 months. The actuarial biochemical recurrence free survival rate at 3 and 5 years was 81% and 74%, respectively. Addition of hormonal therapy to salvage radiotherapy did not alter biochemical recurrence rate (P = 0.56). Univariate analysis revealed that Gleason score of 8 to 10 (P = 0.026) and PSA before salvage therapy greater than 0.24 ng/ml (P = 0.0016) were significant risk factors for biochemical recurrence. On multivariate analysis, PSA before salvage therapy greater than 0.24 ng/ml (P = 0.017) maintained statistical significance. Of 28 patients 3 (11%) experienced late grade 3 toxicity of hematuria. CONCLUSION: Our data suggest that early use of salvage radiotherapy is beneficial for patients with biochemical recurrence after radical prostatectomy. PMID- 19999132 TI - [Clinical factors associated with benign renal tumors]. AB - OBJECTIVES: In this study, we sought to define the incidence of benign renal tumors in our institute and to clarify the clinical factors associated with benign renal tumors, in order to assist in forming preoperative differential diagnoses. METHODS: From October 2002 to July 2007, we performed 157 nephrectomies in patients preoperatively diagnosed with renal cell carcinoma. We chose 81 tumors, all of which were less than 5 cm, for further study. We reviewed double-phase helical CT imaging retrospectively, specifically focusing on attenuation patterns and homogeneity. We also compared clinical factors, including age, sex and tumor size, between the benign and malignant renal tumors. RESULTS: The patient's median age was 67 years (mean age, 63 years), and the median tumor diameter was 3.0 cm (mean, 3.2 cm). Benign renal tumors were found in 10 (12%) of the 81 tumors; these included seven cases of oncocytoma and three cases of angiomyolipoma with minimal fat. Several factors were significant clinical determinants of differentiation between benign and malignant renal tumors: homogeneity in CT, female gender, and small tumor size all predominated in cases of benign tumors. Attenuation pattern in CT, however, was not a significant factor (p = 0.344). CONCLUSIONS: When a patient, especially a female, presents with a small and homogeneous renal tumor, careful consideration should be given to the possibility of a benign process, which needs further consideration before performing excessive surgery. PMID- 19999133 TI - [Clinical efficacy of tolterodine for patients with overactive bladder after insufficient efficacy by monotherapy with alpha1-adrenoceptor antagonist]. AB - PURPOSE: The efficacy of alpha1-adrenoceptor (alpha1-AR) antagonist and anticholinergic agent combined therapy for patients with benign prostatic hyperplasia (BPH) together with overactive bladder (OAB) has been controversial. The purpose of this study was to evaluate the effect of tolterodine combined with alpha1-AR antagonist for patients with BPH and OAB after insufficient efficacy by monotherapy with alpha1-AR antagonist. The adverse event of this combined therapy was also assessed. MATERIALS AND METHODS: The study included 47 patients with BPH, whose OAB symptom persisted (OAB symptom score; OABSS > or =3) after monotherapy with alpha1-AR antagonist for more than 4 weeks. The mean age was 72.9 years and the mean prostate volume was 29.8 ml. Four mg/day of tolterodine with alpha-AR antagonist was administered for 8 weeks to patients. International prostate symptom score (IPSS), quality of life (QOL) index, OABSS, King's Health Questionnaire (KHQ) and residual urine volume (RUV) were assessed before and after combined therapy. RESULTS: Six patients were dropped out from this study because of dry mouth, constipation, onset of other disease and insufficient efficacy by self-judgment. IPSS (from 15.1 +/- 6.8 to 11.0 +/- 7.9; P < 0.01), QOL index (from 4.3 +/- 1.1 to 3.6 +/- 1.3; P < 0.01) and OABSS (from 7.0 +/- 3.0 to 5.4 +/- 2.9; P < 0.01) of 41 patients improved significantly by combined therapy. The storage symptom of IPSS subscore improved significantly (from 8.0 +/ 2.9 to 6.5 +/- 2.8; P < 0.01), whereas the voiding symptom did not improve. Regarding KHQ, the score of 3 domains (impact on life, role limitation, and physical limitation) improved significantly (P < 0.05). RUV did not change and no serious adverse event including urinary retention was found in this study. CONCLUSIONS: This study reveals that the combined therapy of alpha-AR antagonist and tolterodine represents an effective and safe treatment modality for patients with BPH and OAB, whose OAB symptom was not improved by antecedent monotherapy with alpha-AR antagonist. PMID- 19999134 TI - [Ureteral endometriosis in estrogen-treated postmenopausal woman]. AB - A 56-year-old woman was admitted to our hospital due to gross hematuria. Cystoscopy and contrast-enhanced abdominal CT scan revealed a solid tumor at the right terminal ureter. She underwent transurethral resection of the right ureteral tumor. The ureteral tumor included proliferative endometrial stroma and glands under urothelial cells, and the histopathological diagnosis was ectopic endometriosis. Before the surgery, the patient had underwent hormone replacement therapy using estrogen patches to treat menopausal disorders, however, the dose of estrogen was five times higher than regular dose. Therefore, in this case, the serum level of estrogen was elevated over the normal value. Ectopic endometriosis is rare in urinary tract especially ureter. Furthermore, endometriosis is an uncommon disease in women during postmenopausal period. Our case suggests that an excessive hormone replacement therapy might cause endometriosis in postmenopausal women. PMID- 19999135 TI - [Case of malignant lymphoma of the prostate complicated with prostate adenocarcinoma]. AB - A 68-year-old man was referred to our hospital with complaints of palpation, hematemesis and melena. Esophagogastroduodenoscopy revealed a huge ulcer in the stomach, and based on biopsy findings, he was pathologically diagnosed as having diffuse large B-cell type malignant lymphoma. A computed tomographic scan demonstrated prostatic enlargement and swelling of the left external iliac lymph nodes. Since his serum PSA level was 13.0 ng/ml, prostatic needle biopsy was performed. Histological findings revealed diffuse large B-cell type malignant lymphoma and moderately differentiated adenocarcinoma of the prostate. The patient achieved complete response after eight cycles of combination chemotherapy with rituximab cyclophosphamide, adriamycin, vincristine and predonisolone. At the same time of chemotherapy, androgen deprivation therapy was initiated. The current his PSA level is 0.2 ng/ml or less. PMID- 19999136 TI - [Case of primary leiomyosarcoma of the seminal vesicle]. AB - A 72-year-old male visited our hospital with a complaint of perineal and voiding pain. An elastic hard mass was palpated by rectal examination in the cranial lesion of prostate. On CT scan and MRI it was located at the site of the right seminal vesicle. Trans-rectal needle biopsy specimen showed leiomyosarcoma. Radical cystoprostatectomy with pelvic lymph node dissection was performed at Hokkaido University Hospital. Microscopic examination of the resected specimen revealed the tumor arose from the right seminal vesicle without involvement of the bladder and the prostate gland. Three months after the operation the tumor recurred at lung and pelvic lymph node. He died 10 month postoperatively. Autopsy finding showed many visceral organs metastases without local recurrence. In the past, nine cases of leiomyosarcoma of seminal vesicle were published in the literature. Including our case, pathological diagnosis by core needle biopsy was attempted in five cases, though accurate diagnosis was made in only two cases. In our case, Immunohistological study by SMA and desmin was useful for histological diagnosis preoperatively. PMID- 19999137 TI - [Metastatic malignant melanoma of the urinary bladder: a case report]. AB - A 62-year-old male underwent resection of malignant melanoma of left breast skin in 2006. In 2007, he underwent lymph node dissection and chemotherapy (DAV-feron therapy) for left axillary lymph nodes metastasis. In Aug 2008, he was referred to our department with chief complaint of macrohematuria. Two non-papillary bladder tumors were detedted on cystoscopy and CT/ MRI showed multiple lymph node swelling, including left inguinal, paraaortic, and right cervical regions. As malignant melanoma cells were found on urinary cytology, a diagnosis of metastatic malignant melanoma of the urinary bladder was made. It was thought that tumor resection would not contribute to prognostic improvement because of multiple lymph node metastases. Therefore, tumor resection was not performed. The patient is currently receiving chemotherapy. This is the eleventh case of metastatic malignant melanoma of the urinary bladder to be reported in the Japanese literature. There have been no previous reports of cases in which urinary cytology was positive for malignant melanoma cells. PMID- 19999138 TI - [Productive activities and well-being among community-dwelling elderly]. AB - OBJECTIVE: This study examined the association between engagement in productive activities and well-being among community-dwelling elderly. METHODS: Data for 576 older adults were obtained from a mail survey in Osaka City. The productive activities was measured with reference to three domains (paid work, unpaid work at home, and unpaid work outside the home) and the number of productive roles. Well-being was assessed by two measures: life satisfaction (LSIK) and self-rated health. The author used multivariate analyses with each of the two measures of well-being as dependent variables, and each of the productive activities as independent variables. The analyses were conducted separately for men and women, controlling for sociodemographic variables, IADL(instrumental activities of daily living) and social relationships. RESULTS: Multivariate analyses revealed the following findings. Paid work was positively associated with life satisfaction of women and self-rated health of men and women. Unpaid work at home was not significantly associated with life satisfaction or self-rated health for either men or women. However, unpaid work outside the home was related to higher scores for life satisfaction and self-rated health among women. The numbers of productive roles were positively associated with life satisfaction of women and self-rated health of men and women. CONCLUSION: Productive engagement was generally beneficial to older women's well-being except for unpaid work at home. Among men, only paid work out of the three activity domains and the number of productive roles were associated with better self-rated health. Public health policies and programs that bring older women into productive engagement outside the home may thus improve their well-being and generate benefits for communities. PMID- 19999139 TI - [A functional fitness test battery for pre-frail older adults (so-called "specified elderly individuals")]. AB - BACKGROUND: In Japan, an exercise program for pre-frail older adults (so-called "specified elderly individuals") (SEIs) has been prescribed under the newly developed long-term care insurance system. Three functional fitness measures (grip strength, one-leg balance with eyes open, and 5-m habitual walk) are used in the system; however, it has yet to be determined if applying these measures to SEIs is appropriate. PURPOSE: The purpose of this study was to develop a new test battery assessing functional fitness for SEIs, and to determine cross-validity and responsiveness of these measures. METHODS: One hundred and twenty seven SEIs (76.6 +/- 5.9 yr) and 315 healthy older adults (HOAs) (72.2 +/- 5.8 yr) completed twelve functional fitness tests related to activities of daily living (ADL) and mobility. The SEI was defined by the Japanese Ministry of Health, Labour and Welfare in 2005. The test battery items were selected by logistic regression analysis. A functional fitness score (FFS) equation was developed by principal component analysis. The cross-validity of the FFS equation was then tested using a different set of 28 SEIs (77.5 +/- 6.5 yr) and 143 HOAs (71.5 +/- 4.7 yr). Responsiveness of the FFS was also assessed in 62 SEIs (76.7 +/- 5.9 yr) after a 3-month exercise program. RESULTS: The following 4 test items were selected for assessment of functional fitness in SEIs: tandem stance, 5-repetition sit-to stand, alternate step, and timed up and go. Applying principal component analysis to the 4 selected functional fitness items, the first principal component was interpreted as total functional fitness. The following equation was developed to estimate FFS based on the first principal component coefficient of each variable: FFS = 0.031X1-0.106X2-0.192X3-0.096X4 + 1.672, X1 = tandem stance(s), X2 = 5 repetition sit-to-stand (s), X3 = alternate step(s), X4 = timed up and go(s). The cut-off value to distinguish SEIs from HOAs using receiver operating characteristic ROC) curve was 0.065 (sensitivity 82.2%, specificity 81.9%). The cross-validity and responsiveness of the FFS equation was considered acceptable. CONCLUSION This newly developed test battery should be a useful tool for comprehensively evaluating functional fitness in SEIs. PMID- 19999140 TI - [Relationships between perception of Exercise Guidelines 2006, perception of other health promotion policies, and demographic variables]. AB - PURPOSE: According to the interim report of Healthy Japan 21, promoting Exercise Guideline 2006 is one important issue in the fields of physical activity and exercise. However, a previous study indicated that Japanese people rarely were aware of Exercise Guideline 2006 (Higo & Nakamura, 2008). The purpose of the present study was to identify characteristics of those who were aware of the guideline. METHODS: The subjects were 1,613 Japanese adults (40.2 +/- 12.2 years) recruited by a Japanese social research company. The study designwas cross sectional, using an internet questionnaire. Regarding the research items, the dependent variable was awareness of Exercise Guideline 2006, and the independent variables included awareness of other health promotion policies such as Healthy Japan 21, the Japanese Nutrition Balance Guide, and the Japanese national policy for preventing metabolic syndrome, as well as demographic variables (e.g., gender, age, BMI, hours per day spent watching TV watching and hours per day with internet usage). Logistic regression analysis was used for this research. RESULTS: Of all subjects, 12.3% answered that they were aware of Exercise Guideline 2006. Those over 50 years old (OR = 2.17; 95% CI = 1.11-4.22), having over 10 million household income (OR = 1.94; 99% CI = 1.05-3.61), an exercise habit (OR = 1.75; 95% CI = 1.07-2.86), an awareness of Healthy Japan 21 (OR = 23.60; 95% CI = 15.26-36.52), the Japanese Nutrition Balance Guide (OR = 5.52; 95% GCI = 3.01-10.13), and the Japanese national policy for preventing metabolic syndrome (OR = 3.41; 95% CI = 2.12-5.48) were significantly more likely to be aware of the guideline. CONCLUSIONS: Major findings of this study were: 1) older people are more aware of Exercise Guideline 2006 than their younger counterparts, although level of awareness was generally low; 2) for promotion of Exercise Guideline 2006, targeting those with a low educational background might not be a critical issue; and 3) awareness levels for Exercise Guideline 2006 and HealthyJapan 21 are similar. Development of effective and specific ways for promoting Exercise Guideline 2006 are a high priority. PMID- 19999141 TI - [Analyses of influenza epidemics in the 2006/2007 season and vaccine effectiveness in a local town in Oita Prefecture]. PMID- 19999142 TI - [Basic mechanism of magnetic human cerebellar stimulation and its clinical application]. AB - I here summarize the history of cerebellar stimulation experiments in humans and give some caution to use this stimulation method. In clinical evaluation, we consider the cerebellum as a kind of computer to get information from the peripheral structures and also higher motor cortical centers including the primary motor cortex (M1) and send a cerebellar command to M1 after computation of much information. We study functions of the cerebello-afferent and cerebello efferent connections using cerebellar stimulation and differentiate these pathways dysfunction. We first activated the cerebellum using electrical stimulation. The most effective position, effective current direction and the interval of conditioning and test stimuli suggested that the observed effect might be produced by some cerebellar structures activation. Studies of cerebellar ataxia patients and other disorders supported the idea that the suppression is produced by the inhibition of dentato-thalamo-cortical pathway by Purkinje cell activation. In patients with a lesion at cerebellar hemisphere, dentate nucleus, superior cerebellar peduncle, motor thalamus, the suppression effect was not evoked. In contrast, the suppression was normally elicited in patients with a lesion at pontine nucleus, middle cerebellar peduncle even though they had clinically definite ataxia. Normal suppression was evoked in patients with non cerebellar ataxia (sensory ataxia due to paraneoplastic syndrome, tabes dorsalis, ataxic sensory neuropathy). Based on these results, we concluded that the cerebellar electrical stimulation method was useful to differentiate cerebellar ataxia due to cerebellar efferent pathways lesions from other cerebellar ataxia and non-cerebellar ataxia. We demonstrated that magnetic stimulation over the cerebellum using a double-cone coil can produce the same effect as those elicited by electrical cerebellar stimulation. These all results supported the proposal that the magnetic stimulation over the cerebellum can enable us to differentiate the cerebellar efferent ataxia from other cerebellar ataxia and non-cerebellar ataxia. A recent paper has cautioned us to conclude the observed phenomenon to be produced by cerebellar activation after exclusion of several other factors as stated in the original paper. The most serious factor to exclude is the antidromic activation of the corticospinal tracts by the cerebellar stimulation conditioning stimulus. To exclude this possibility, it is important how to measure the threshold of the corticospinal tracts. We recommend that we should use rectified EMG recordings when determining it. In summary, I conclude that the cerebellar magnetic stimulation is a good tool for physiological differentiation of cerebellar ataxia mechanisms in ataxic patients. At a current stage, I recommend a conservative method mentioned in the editorial paper for magnetic cerebellar stimulation. PMID- 19999143 TI - [Relationship between left atrial size and paroxysmal atrial fibrillation in acute ischemic stroke]. AB - We investigated the relationship between parosysmal atrial fibrillation (pAF) and left atrial (LA) size in patients with acute ischemic stroke. Between June 2006 and April 2008, we retrospectively enrolled 292 patients with acute ischemic stroke within 24 hours of onset, who measured LA size by transthoracic echocardiography. The patients were classified according to the presence or absence of chronic AF on admission (cAF and normal sinus rhythm (NSR) group). The NSR group was subdivided based on the pAF (pAF and non-AF group). We compared LA size among each groups. Furthermore in the NSR group, the factors associated with pAF were investigated by multivariate logistic regression analysis. Among the enrolled patients, cAF (cAF group) had 77 (26.4%), pAF (pAF group) had 32 (11.0%) and non-AF group was 183 (62.7%). The median of LA size of the cAF was highest (4.7 cm), followed by the pAF group (4.1 cm) and the non-AF group (3.5 cm) (p<0.001). Median age (72.0 for the non-AF group vs. 74.5 years for the pAF group, p<0.001), NIHSS score on admission (3.0 vs. 12.5, p<0.001), D-dimer (0.6 vs. 2.1 microg/ml, p=0.003), LA size (3.5 vs. 4.1 cm, p<0.001) were higher in the pAF group than in the non-AF group. The optimal cut-off value, sensitivity and specificity of LA size to distinguish pAF from non-AF were 3.8 cm, 68.6% and 73.8%, respectively. Multivariate logistic regression analysis demonstrated that a NIHSS score of > or =8 (odds ratio [OR], 4.399; 95% confidence interval [CI], 1.701 to 11.378, p=0.002), LA size of > or =3.8 cm (OR, 8.882; 95% CI, 3.238 to 24.268, p<0.001) and mitral valvular disease (OR, 4.677; 95% CI, 1.720 to 12.720, p=0.003) were independent factors associated with pAF. We should consider the presence of pAF when LA size is over 3.8 cm in acute ischemic stroke patients with sinus rhythm. PMID- 19999144 TI - [An acute severe heat stroke patient showing abnormal diffuse high intensity of the cerebellar cortex in diffusion weighted image: a case report]. AB - A 47-year-old man was admitted to the hospital because of general convulsion, loss of consciousness and hyperthermia. A diagnosis of acute heat stroke was made clinically and neuroradiologically. As the consciousness level ameliorated, he developed severe abulia and mutism, then cerebellar ataxic syndrome (viz. truncal ataxia, hypermetria, ataxic speech and nystagmus). An MRI (diffusion weighted image; DWI) disclosed abnormal diffuse high signal intensity of the cerebellar cortex with reduced apparent diffusion coefficient (ADC). Two months later after the onset, truncal ataxia and dysarthria significantly improved, while dysmetria of the extremities rather worsened. At that time, the abnormal signal intensity of the cerebellar cortex disappeared, and the cerebellum became atrophic. The cerebellar blood flow was significantly decreased on brain SPECT (99mTc-ECD). The abnormal DWI signal intensity of the cerebellar cortex in the present patient may represent the cytotoxic edema of Purkinje cells resulting from heat stroke related hyperthermia It is essential to repeat MRI examination for cerebellar pathology and to obtain better insight into sequelae in patients with acute heat stroke. Protirelin tartrate seemed to be valid for improvement of abulia in the present patient. Further study is indicated. PMID- 19999145 TI - [Successful treatment of recurrent hypoglycemia by pioglitazone in a patient with myotonic dystrophy]. AB - A 20 year-old woman with myotonic dystrophy type 1 (DM1) presented with fatigue, daytime somnolence, and sudden poor responsiveness. Blood glucose was measured before and after each meal for 4 days, and hypoglycemia was confirmed twice, although neither perspiration nor palpitations occurred in the hypoglycemic state. On a 75 g oral glucose tolerance test (OGTT), fasting blood glucose level was 83 mg/dl, and fasting blood immunoreactive insulin (IRI) level was 5.96 microIU/ml. However, IRI increased to 528 microIU/ml at 60 minutes and blood glucose decreased to 57 mg/dl at 120 minutes of the OGTT. The patient was diagnosed with reactive hypoglycemia due to excessive insulin secretion. Oral administration of pioglitazone improved the excessive insulin secretion as assessed by OGTT. After starting treatment, hypoglycemia was not detected either pre- or post-prandially. After 10 months of treatment, blood glucose level after glucose loading was higher than fasting blood glucose level during OGTT, and the IRI area under the curve of the OGTT decreased. We considered that hypoglycemia unawareness resulted from recurrent hypoglycemic episodes in this patient. Pioglitazone was effective in improving hyperinsulinemia and reactive hypoglycemia in nondiabetic DM1. PMID- 19999146 TI - [An advantage of T2*-weighted MRI for early detection of straight sinus thrombosis: a case report]. AB - A 44-year-old man presented with a 12-day history of severe non-throbbing headache. He showed no physical abnormality but obesity. On day 12, ring-shaped low intensity lesions inside straight sinus were revealed on T2*-weighted MRI image (T2*WI). On the following day (day 13), he was found unresponsive at home, and ambulated with disturbed consciousness. FLAIR and diffusion-weighted MRI image disclosed high intensity signals in bilateral thalamus which were postulated as vasogenic edema. MR venography and conventional cerebral angiography showed an absence of flow in inferior sagittal sinus, vein of Galen, and straight sinus. These findings confirmed the diagnosis of cerebral venous thrombosis (CVT). Anticoagulant treatment was introduced and his consciousness level was gradually improved. On day 43, he was discharged with no neurological sequelae. A delay of correct diagnosis and treatment with CVT can lead to devastating disability or even to death. An early diagnosis of CVT is often dismissed owing to the nonspecific symptoms such as headache and nausea. Recent reports described high sensitivity of T2*WI for detecting CVT. Alterations in blood flow and oxyhemoglobin reduced products, deoxyhemoglobin, in thrombosed veins often produce the magnetic susceptibility on T2*WI. A detection of ring shaped low intensity lesions within venous sinus on T2*WI were quite rare, and the signal changes of these sinus lesions were successfully visualized by chronological T2*WI. Taken together, our case implies that T2*WI is the powerful tool for the early detection of CVT, even before the critical symptoms might happen. PMID- 19999147 TI - [A case of cerebellar syndrome associated with HIV infection]. AB - A 36-year-old man was hospitalized because of subacutely progressive gait disturbance. Neurological examination disclosed severe ataxia of gait and trunk and moderate ataxia of the four limbs, without signs of cognitive impairment. There were no manifestations of systemic infections. Brain MRI showed mild atrophy of the cerebellar vermis and hemispheres. Extensive laboratory search failed to disclose the cause of subacute ataxia. Cerebellar ataxia progressed, leading to the patient becoming wheelchair-bound two months after admission, when PCR analysis of the cerebrospinal fluid was positive for Epstein-Barr, JC, and hepatitis B viruses. In addition, the quantity of serum HIV1-RNA was 2.9 x 10(4) copies, the absolute count of CD4+ lymphocyte was 28/mm3, and the CD4/CD8 ratio was 0.04, despite clear denials by both the patient and his wife regarding any apparent infectious opportunities. Accordingly thereafter, highly active antiretroviral therapy was initiated. Several weeks after the initiation of therapy, ataxia stabilized with disappearance of serum HIV and cerebrospinal fluid JCV viral load. He returned to his occupation 20 months after disease onset without progression of ataxia or development of other neurological dysfunctions including dementia. We could not establish the exact pathogenesis of ataxia in this patient It could have been primary cerebellar degeneration caused by HIV, or the other viruses detected (EBV, JCV) or autoimmune mechanisms caused by these viruses. However, HIV infection should be considered as an etiology in clinical setting of subacute ataxia, particularly in a young or immunocompromised patient. PMID- 19999148 TI - [A case of neurosarcoidosis with recurrent episodes of neurological dysfunction and diffuse cortical lesions on magnetic resonance imaging]. AB - We report a case of a 35-year-old man with histologically confirmed neurosarcoidosis who developed recurrent episodes of right-hemispheric dysfunction with diffuse cortical lesions of the right hemisphere on magnetic resonance imaging (MRI). A brain biopsy revealed granulomatous inflammatory cells in both the subarachnoid space and Virchow-Robin space, which might relate to the recurrent neurological dysfunction and MRI findings. PMID- 19999149 TI - [Effects of acetylcholinesterase inhibitors on patients with MuSK antibody positive myasthenia gravis]. AB - In order to clarify the clinical characteristics and effects of acetylcholinesterase inhibitors of patients with generalized myasthenia gravis with antibodies to muscle specific kinase (MuSK), we investigated seven patients with MuSK antibodies and eleven patients without both antibodies of acetylcholine receptor and MuSK. All patients with MuSK antibodies showed bulbar symptoms, which frequency was significantly higher compared to those in patients without double antibodies. The frequency of positive result of Tensilon test was significantly lower in patients with MuSK antibodies than in those without double antibodies. In response to intravenous edrophonium chloride, MuSK positive patients showed adverse reactions in a small dosage of edrophonium chloride, less than 5 mg, such as fasciculation on facial muscles and stuffy sensation of throat. The adverse responses to a small dosage of intravenous edrophonium chloride injection is useful information to distinguish patients with seronegative generalized MG, whether they have MuSK antibodies or not. When acetylcholinesterase inhibitors medication is tried to patients with MuSK antibodies, if necessary, a small dosage of inhibitors should be used to avoid cholinergic hypersensitivity. PMID- 19999150 TI - [A case of Castleman disease accompanying neuropathy only detected by S-SEP in the tibial nerve]. AB - A 77-year-old woman was admitted with a chief complaint of tingling sensation in the both feet, which gradually developed just after the diagnosis of Castleman disease was made. Neurological examination showed mild weakness in the neck and pelvic girdle muscles, and sensory impairment affecting all modalities in the lower legs. Although these neurological findings suggest a diagnosis of neuropathy, nerve conduction studies (NCS) and F-wave disclosed no abnormalities. However, the short latency somatosensory evoked potential (S-SEP) in the tibial nerve revealed a significant delay in the P15 latency, which is indicative of neuropathy affecting proximal potion of the peripheral nerve. To our knowledge, only a few reports described proximal neuropathy associated with Castleman disease. In our case, the examination of S-SEP was very informative to make a diagnosis of neuropathy. PMID- 19999151 TI - [Electrocautery snare efficacy in head and neck lesion treatment]. AB - The electrocautery snare has been widely used in gastroenterology and to remove bronchial and urinary bladder lesions, but rarely in head and lesion electrocautery. Since October 2006, we have used this instrument to treat 17 head and neck granuloma, papilloma, cyst, and cancer lesions under local or general anesthesia. Lesions were localized in the larynx, oropharynx, hypopharynx, or tracheostoma. The snare was used through a rhinolaryngovideoscope in most cases with a current of 15-30 watts produced by an electrosurgical generator. All procedures were easy, quick and successful, with minimal bleeding. No severe adverse effects were seen in any of our cases. The electrocautery snare was extremely useful in treating pedunculated lesions and in removing epiglottic cysts with a clear margin after excision of the mass without bleeding. Our results indicate the electrocautery snare to be useful and safe in treating selected head and neck lesion cases. PMID- 19999152 TI - [A case of difficult-to-diagnose hereditary angioedema]. AB - Hereditary angioedema (HAE) due to an inherited C1-inhibitor (C1-INH) deficiency causes localized swelling of the oral cavity, pharynx, larynx, and face, that may be life-threatening when the larynx is involved. A 26-year-old woman seen 3 times previously for pharyngeal or laryngeal edema while in her teens, and seen this time for dyspnea was found in computed tomography (CT) to have esophageal edema and pleural effusion. Her C1-INH activity was low, yielding a definitive diagnosis of HAE for her 10-year-plus-disease history. n mind in th e f idiopathicedema. should be kept i While it is comparatively rare, HAE differential diagnosis o PMID- 19999153 TI - [A case of Langerhans cell histiocytosis coincident with severe external ear canal inflammation]. AB - We report a case of Langerhans cell histiocytosis (LCH), a rare disease caused by the proliferation of abnormal Langerhans cells, coincident with severe external ear canal inflammation. A 27-year-old man with a 1-year history of external ear canal discharge was found in computed tomography (CT) to have left temporal bone erosion with tissue granulation. Chest X-ray showed pulmonary fibrosis, necessitating transbronchial lung biopsy that yielded a definitive diagnosis of multisystem, multisite LCH involving the bone, skin, lung, pituitary, thyroid, and lymph node systems. He underwent combination chemotherapy directed by the Japan LCH Study Group. LCH should therefore be considered in the case of a patient with severe external ear canal inflammation coincident with temporal bone erosion. PMID- 19999154 TI - [Bioimaging for the study of trans-hierarchical interplay in biological systems]. PMID- 19999155 TI - [Visualisation of conformational changes of proteins at the single-molecule level]. PMID- 19999156 TI - [Single-molecule imaging analysis of stochastic signal transduction in chemotactic response]. PMID- 19999157 TI - [Imaging of intracellular ATP using novel fluorescent probes]. PMID- 19999158 TI - [Ultra-sensitive and large-scale imaging of cellular coupled oscillator systems]. PMID- 19999159 TI - [Visualization of neuronal network activity]. PMID- 19999160 TI - [Physical simulation of the transcriptional network of mammalian circadian clocks]. PMID- 19999161 TI - [Discovery and assembly mechanism of degenerated subunits in succinate dehydrogenase complex]. PMID- 19999162 TI - [Novel transcriptome analysis method incorporating multi-sample profile correlation: its application to glycan biosynthetic pathways]. PMID- 19999163 TI - [Practical guide in theoretical biology for molecular biologists]. PMID- 19999164 TI - Role for netrin-1 in sensory axonal guidance in higher vertebrates. AB - During development, dorsal root ganglion (DRG) neurons in higher vertebrates extend their axons centrally to the spinal cord through the dorsal root entry zone (DREZ) and peripherally to muscle and skin targets. After entering the spinal cord, DRG axons project into the dorsal mantle layer. In this review, we focus on evidence showing the role for netrin-1 in forming sensory axonal trajectories. Netrin-1 is a diffusible axonal guidance molecule that chemorepels developing DRG axons. When DRG axons project toward the DREZ, ventral spinal cord derived netrin-1 prevents DRG axons from projecting aberrantly toward the ventral spinal cord. At later stages, the dorsal spinal cord cells transiently express netrin-1. This dorsal spinal cord-derived netrin-1 prevents DRG axons from invading the dorsal spinal cord during the waiting period. Together, the data reviewed provide strong evidence that netrin-1 plays a crucial role in sensory axon projection during development. PMID- 19999165 TI - Separate demonstration of arterial- and venous-phase by 3D-CT angiography for brain tumors using 64-multidetector row CT: 3D-CT arteriography and 3D-CT venography. AB - We assessed the usefulness of the separate demonstration of the arterial- and venous phase on 3D-CT angiography (3D-CTA) using a 64-multidetector row CT (MDCT) scanner for the surgery of brain tumors. Nineteen patients with meningiomas (n=11), schwannomas, metastatic brain tumors (n=2 each), glioblastoma multiforme, malignant lymphoma, craniopharyngioma, and embryonal carcinoma (n=1 each) underwent scanning on a 64-MDCT scanner. After dynamic CT scanning to determine the scan timing for the arterial- and venous-phase, we individually scanned the arterial- and venous phase for 4 sec after injecting a nonionic contrast medium. Using the CT threshold setting and subtraction and cutting techniques, we produced individual 3D-CT images of the arteries, veins, tumors, and bones. The operators subjectively assessed the usefulness of these images in comparison with 3D-CTA. We separately demonstrated the arterial- and venous phase on 3D-CTA covering the entire head in all 19 cases. The 3D-CT arteriographs, 3D-CT venographs, and the fused 3D-CT images facilitated our understanding of the 3D anatomic relationship among the tumor, arteries, veins, and bony structures. In 14 of 19 cases our method provided the surgically valuable findings; the information on the anatomical relation between tumor and the surrounding arteries and veins (in 13 cases) the identification of anatomical course of the encased vessels (in one), and feeding arteries and draining veins (in one), and discrimination between the venous sinus and tumor (in one). The anatomical information yielded by our technique makes safer surgery possible. If more detailed information which 3D-CTA cannot provide is required, our method should be performed. PMID- 19999166 TI - Synthetic oligodeoxynucleotides induce gastritis in mice. AB - To investigate whether DNA directly induces gastritis and/or peptic ulcer, we injected synthetic DNA including CpG motif (CpG-DNA) to mouse stomach. BALB/c mice were injected with either saline, acetic acid (AA), CpG-DNA, or Control-DNA. Mice were sacrificed, and sections of the stomachs were stained with hematoxylin and eosin. The lesions were histopathologically scored from 0 to 4 based on the extent of the inflammation. Populations of neutrophils and mononuclear cells infiltrated to the lesion were calculated. IFN-y mRNA expression at the injection site was analyzed by RT-PCR. The number of CpG motifs included in the complete genomes of H. pylori HP26695 and J99, Escherichia coli O157, and Salmonella Typhi was determined by genomic analysis of these bacteria. Intragastric injection with CpG-DNA induced gastritis, and statistical analysis of histological scores revealed a significant difference between saline vs CpG-DNA (p = 0.037). The population of mononuclear cells infiltrated to the lesions was significantly higher in mice injected with CpG-DNA than that injected with AA (p = 0.0061). IFN gamma mRNA expression was detected in the CpG-DNA group. While H. pylori includes multiple CpG motifs in its genome, it has fewer than the other pathogenic gram negative bacilli. We conclude that synthetic DNA including CpG motif directly causes gastritis in mice and induces IFN-gamma production in the stomach. Bacterial DNA including CpG motif is known to stimulate innate immunity and to cause inflammation. Thus, H. pylori genomic DNA may be one of the virulent factors involved in H. pylori infection. PMID- 19999167 TI - Cochlear implantation in a case of bilateral sensorineural hearing loss due to mumps. AB - It is well known that unilateral profound sensorineural hearing loss is caused by mumps; however, bilateral deafness is rare. Herein we report a case of bilateral profound hearing loss caused by mumps infection in a four-year-old boy. Labyrinthitis due to the mumps virus was suspected. His verbal understanding was poor, and he completely stopped talking. He was soon fitted with a hearing aid, but it proved insufficient. Thereupon, cochlear implantation was performed on his left ear. Six months after the operation, his speech perception and speech production were improved. In cases of bilateral profound hearing loss due to mumps infection conservative therapy is ineffective; therefore, cochlear implantation is recommended. Vaccine coverage for mumps virus is also strongly recommended in Japan. PMID- 19999168 TI - The challenging economics of US health care reform. PMID- 19999169 TI - PRODUCT: Embeda 20mg/0.8mg. PMID- 19999170 TI - Is this skin diseased, or is it just dirty? PMID- 19999171 TI - Major depression. Screening and treatment. PMID- 19999172 TI - The role of human papillomavirus testing in cervical cancer screening. PMID- 19999173 TI - An uncommon cause of mechanical small bowel obstruction. PMID- 19999174 TI - Have the latest CPR guidelines improved cardiac arrest outcomes? PMID- 19999175 TI - Surgical treatments for patients with an infected total knee arthroplasty. PMID- 19999176 TI - Managing the adverse effects of bisphosphonate therapy on the jaw. PMID- 19999177 TI - Lipid disorders. PMID- 19999178 TI - The art and science of defensive medicine. PMID- 19999179 TI - Foot pain for 5 years, and an abnormal radiograph. PMID- 19999181 TI - Patient information. how can I reduce the risk of SIDs? PMID- 19999180 TI - When the patient asks. How can I reduce the risk of SIDs? PMID- 19999182 TI - Helping your patients to deal with a predisposition to genetic disease. PMID- 19999183 TI - Case of the month. Diagnosis: Meatal stenosis. PMID- 19999184 TI - [Copernican revolution in the therapy of rheumatoid arthritis: the contribution of anti-TNFalpha drugs]. AB - TNFalpha has a key role in cell recruitment, proliferation and death, expression of adhesion molecules and immune responses. In RA, TNFalpha is involved in matrix degradation and osteoclastogenesis. TNFalpha inhibitors are either soluble receptors (etanercept) or monoclonal antibodies (infliximab and adalimumab; golimumab and certolizumab are in development). TNFalpha antagonists, alone or in combination with methotrexate, reduce bone erosions and thinning of cartilage, but they differ as regards ligand binding, pharmacokinetics, pharmacodynamics, and clinical indications. Etanercept is the only TNFalpha antagonist that also neutralises LFT-alpha. Infliximab and adalimumab are more immunogenic. Cytotoxicity and cellular lysis are also higher with infliximab and adalimumab. Etanercept slows progression of joint damage in recently diagnosed RA when given alone, but much more when given with methotrexate; anti-TNF monoclonal antibodies also were shown to slow progression alone and in combination with methotrexate. Patients with early and long-standing RA treated with etanercept have now shown improvement in ACR scores, inflammation and disability for up to 9 years. Outcomes with infliximab and adalimumab are similar to those with etanercept, but only in combination with methotrexate. As a result of neutralizing antibodies, increasing doses of anti-TNFalpha antibodies may be required to maintain clinical response. As regards side effects, opportunistic infections seem more frequent with monoclonal antibodies. TNFalpha antagonists produce more QALYs than traditional DMARDs, counteracting higher costs. The efficacy, safety, and quality of life benefits of TNFalpha antagonists suggest using them possibly earlier than today, even in clinically moderate RA. Thanks to its overall profile, etanercept might be considered as one of the first-choices in TNFalpha antagonism in RA management. PMID- 19999185 TI - [Observational studies in osteoporosis treatment]. AB - Osteoporosis is characterized by increased fracture risk, with associated severe morbidity and mortality. Several pharmacological treatments have been demonstrated to be effective in reducing fracture risk, by means of Randomized Controlled Trials (RCTs). However, it seems that the results of these RCTs could not be immediately generalized to the "real-world" population. It has been calculated that less than 20% of the subjects included in the RCTs may be representative of the general population treated for osteoporosis in clinical practice, due to the very strict inclusion and exclusion criteria of these trials. In the last years, some observational studies on osteoporotic-treated patients have been carried out in those Countries where wide administrative claims database are available. These studies may be useful because of the large "real-world" population sample included, the possibility to control for several comorbidities and active treatments, and the opportunity to follow patient outcome over time, even comparing the clinical effects of the different drugs used for fracture prevention. However, their results should be considered with caution, due to the potential bias in the diagnosis of fracture, adherence assessment, type of treatment followed by the patients, and the impossibility to control for several characteristics influencing bone health as well as treatment outcomes. As discussed in the paper, the results of these studies tend to confirm, at least in some cases, the results of the RCTs for fracture prevention, confirming that the drugs currently used for osteoporosis treatment are actually effective in clinical practice as well. PMID- 19999186 TI - Fixed-combination monotherapy with clindamycin 1%- benzoyl peroxide 5% gel: focus on acne patient benefits and clinical use. Introduction. PMID- 19999187 TI - Rapid and efficacious fixed-combination monotherapy: desired results for the patient and improved adherence for the clinician. AB - Ideally, acne therapy should treat multiple causative disease factors and improve clinical symptoms while considering patient tolerability and expectations as well as other factors that could negatively impact patient adherence to therapy. This article reviews data describing the role of inflammation in disease pathogenesis and how it impacts treatment choice. Pivotal efficacy data supporting use of a fixed-combination monotherapy gel containing clindamycin 1% and benzoyl peroxide 5% (C/BPO) also are reviewed. This fixed-combination monotherapy demonstrates efficacy for the treatment of both inflammatory and noninflammatory acne lesions, both alone and in combination with a retinoid, which may favorably influence a patient's adherence to therapy. PMID- 19999188 TI - Improving treatment outcomes: tolerability of a fixed-combination clindamycin 1% benzoyl peroxide 5% hydrating gel as monotherapy or in combination with a retinoid. AB - Patient adherence to acne therapy is a fundamental element of successful treatment outcomes, which can be increased by the selection of rapidly acting, well-tolerated medications used in simplified regimens. A fixed-combination monotherapy gel containing clindamycin 1% and benzoyl peroxide 5% (C/BPO) has an established safety and efficacy profile through almost a decade of published literature and clinician experience. This unique product may improve a patient's adherence to therapy by providing a well-tolerated and efficacious once-daily monotherapy with multiple mechanisms of action. This article reviews data outlining the enhanced tolerability of the fixed-combination C/BPO gel both alone and when used with a retinoid. The favorable tolerability profile of C/BPO hydrating gel may impart additional benefit to the patient in helping to maintain adherence to treatment. PMID- 19999189 TI - Fixed-combination clindamycin 1%-benzoyl peroxide 5% hydrating gel: a flexible component of acne management. AB - This article reviews various studies supporting the use of a fixed-combination monotherapy hydrating gel containing clindamycin 1% and benzoyl peroxide 5% (C/BPO), which is the only available formulation with a hydrating gel vehicle containing a humectant and an occlusive. The C/BPO hydrating gel provides a flexible and complementary efficacy and/or tolerability profile when used alone or with topical retinoids, which results in rapid response in inflammatory and noninflammatory acne. It also mitigates the irritation associated with disease flare or topical retinoid use, and reduces the postinflammatory hyperpigmentation (PIH) seen in women and in patients with skin of color with acne. These benefits are important because they have the potential to improve patient adherence to therapy and clinical outcomes. PMID- 19999190 TI - Nitrous oxide--time to say goodbye! PMID- 19999191 TI - Challenges in access to and utilization of reproductive health care in Pakistan. AB - BACKGROUND: Pakistan's maternal mortality rate is high, and adequate and timely emergency services could prevent most maternal deaths. A woman's right to life saving services of skilled health care providers in childbirth is undeniable. This paper examines factors restricting women's access to emergency obstetric care services in Pakistan. METHOD: This cross-sectional survey on emergency obstetric care services collected information at the health facility level using UN process indicators. The study enrolled 170 health facilities from nineteen randomly selected districts in Punjab and NWFP. RESULTS: Diverse factors limit women's access to Emergency Obstetric Care (EmOC) services. EmOC services were unavailable in most health facilities surveyed. Staff absenteeism, geographic remoteness, delayed access, and ambulance shortages jeopardize the transferral of seriously ill patients to higher level care facilities. Cultural norms dictate that women should be examined by women doctors, whose dearth makes these services inaccessible. CONCLUSION: Many maternal deaths would be avoidable if EmOC health services were accessible. The geographic obstacles to timely access, poor hospital infrastructure, and high staff absenteeism rates require immediate attention. Health facilities' working hours were inconsistent with the provision of around-the-clock essential services, depriving and endangering the lives of many in need. It is imperative to increase skilled female workers capable of managing EmOC problems through proper incentives. A focused approach at local levels through proper supervision, motivation, and management would unquestionably save women's lives. PMID- 19999192 TI - Effects of amlodipine on serum testosterone, testicular weight and gonado-somatic index in adult rats. AB - BACKGROUND: Calcium ions are vital in many biological processes including hormonal secretion, mitosis, reproduction, fertility and regulation of gene expression. Thus, calcium antagonists who are frequently prescribed for the cure of cardiovascular diseases may affect any of these physiological processes. METHODS: This quasi experimental study was done at Army Medical College, Rawalpindi from October 2007 to March 2008. Thirty male Sprague Dawley rats, purchased from National Institute of Health, Islamabad were divided into group A (vehicle treated controls) and group B (amlodipine treated). After receiving drug treatment for 50 days, all the rats were sacrificed. Serum was stored to measure testosterone by enzyme immunoassay technique. The testes were removed for measuring absolute testicular weight and gonado-somatic index. RESULTS: Serum testosterone, absolute testicular weight and gonado-somatic index were found to be significantly reduced in amlodipine treated rats. CONCLUSION: These results suggest that long-term treatment with amlodipine might be associated with significant testicular regression and reduction in serum testosterone. Furthermore, since dihydopyridine antagonists are widely used in hypertension, our data may have some clinical implication in the management of infertility associated with hypertension. PMID- 19999193 TI - Clinicohistological pattern of ovarian tumours in Peshawar region. AB - BACKGROUND: Ovarian tumours are one of the major health problems confronting the general practitioners in general and gynaecologists in particular. Ovarian tumours may either be asymptomatic, found on the routine ultrasound examination or symptoms may be vague till the patient has an acute emergency like torsion or rupture of a benign cyst. The worst is late presentation of a malignant ovarian tumour. There is marked variation in the presentation of the tumour as well as in histological types. This study was undertaken to analyse modes of presentation and various histopathological patterns of ovarian tumours. METHODS: This study was conducted from 1st January, 2002 to 31st December, 2002, in Gynaecology 'A' Unit, Lady Reading Hospital (LRH) Peshawar. After admitting patients with ovarian tumours a detailed case history was taken followed by thorough clinical examination. All the relevant details were recorded using the questionnaire. Patients were investigated after performing various surgical procedures; the specimens of ovarian tumours were subjected to Histopathological examination in the histopathology section, Lady Reading Hospital, Peshawar. RESULTS: Amongst the total numbers of 5732 gynaecological admissions during study period the total numbers of ovarian tumours were sixty-eight. Out of which benign ovarian tumours were 61 (89.71%) and malignant ovarian tumours were 7 (10.29%) There were no tumours with borderline malignancy. The commonest histological pattern observed in the study was epithelial tumours (76.5%) including both benign and malignant tumours. The commonest benign tumour was serous cyst adenoma (24%) followed by mature cystic teratoma (18%). Common malignant ovarian tumours were granulosa cell tumours and Endometriod carcinoma (each 28.5%). CONCLUSION: Epithelial tumours are the commonest variety of ovarian tumours followed by Germ cell tumours. The histological type of ovarian tumour correlates with the prognosis of the tumour. PMID- 19999194 TI - Clinical spectrum and management of ovarian tumours in young girls up to 20 years of age. AB - BACKGROUND: Ovarian tumours are one of the common malignancies all over the world affecting all age groups. This study analyses different clinical presentation and management of ovarian tumours in young girls up to 20 years. METHODS: Patients up to 20 years of age admitted with the diagnosis of ovarian tumour were included. Data collected on a proforma. Variables studied included age, presenting symptoms, investigations, surgical findings, type of surgery, histopathology reports and follow-up. The patients with malignant ovarian tumour were followed by oncologist as well as gynaecologist. Data analysis was done on SPSS. RESULTS: The mean age was 17.27 +/- 2.46 SD years. The common symptoms included abdominal mass, abdominal pain, urinary problems, menstrual irregularities and generalized malaise. All patients were operated after preliminary investigations. Patients were advised to have follow-up post-operatively after 1 month. The follow up was done by oncologist and gynaecologist. Six patients (12.5%) died and 22 (45.83%) were lost to follow up. CONCLUSION: Ovarian tumours are quite common in young girls. Majority of patients seek medical advice once the disease becomes symptomatic, complicated or advanced disease in the case of malignancy. Histopathology of the tumours revealed that epithelial cell tumour is the commonest tumour in contrast to germ cell tumour as reported by world literature. PMID- 19999195 TI - Adolescent ovarian tumors: a clinicopathlogical review of 15 cases. AB - BACKGROUND: To prospectively review the clinicopathologic pattern and out come in 15 patients with adolescent ovarian tumours in Military Hospital Rawalpindi between Jan 2004 to Sep 2007. METHODS: All cases of adolescent ovarian tumours reporting to the gynaecology department of Military Hospital Rawalpindi during the period of enrolment. Out of 15 cases 12 were managed surgically. Data was recorded regarding clinical presentation, patient's age, size of tumour, bilaterality, histopathology, staging if tumour was malignant and sites of extra ovarian involvement. All patients were followed up for one year. RESULTS: Majority of patients fell in the subgroup 14-16 year age. Majority harboring ovarian malignancy belonged to subgroup 17-19 years. Clinical presentation in the majority was mass abdomen and abdominal distension. Approach was transcutaneous in 3 (20%), laparoscopic in 2(20%), and open laparotomy in 10 (60%). Frozen section was performed in 3 cases. Histopathology was benign in 11 cases and malignant in 4. In 3 cases cyst fluid was negative for malignancy. In 2 cyst wall biopsy and one patient where entire cyst was removed histopathology revealed benign serous cyst adenoma/luteal cyst (26%). There were 5 cases of dermoid cysts (33.3%), one patient had bilateral dermoids, one malignant and one benign. All 4 malignancies were found to be to non-epithelial on histopathology. One case had surgery twice. Unilateral oopherectomy followed by recurrence and total abdominal hysterectomy and contralateral salpingo-oopherectomy after 2 years. CONCLUSION: This study shows the preponderance of non-epithelial tumours and high percentages of malignant germ cell tumours in adolescents. The incidence of malignant tumours in adolescents is higher than in adults. Early correct diagnosis could be reached by careful physical examination, imaging and tumour markers Surgery should as much as is safely possible be fertility preserving. PMID- 19999196 TI - Pattern of breast cancer: experience at Lady Reading Hospital, Peshawar. AB - BACKGROUND: Breast Cancer is the commonest malignancy of females all over the world and second leading cause of death due to cancer among females. The aim of this Descriptive study was to see the various features of breast cancer in order to know the pattern of disease in the recent time. The study was conducted from Jan. 2007 to Dec. 2007 in Surgical C Unit, Postgraduate Medical Institute, Lady Reading Hospital, Peshawar, Pakistan. METHODS: Study included all patients presenting to and admitted in Surgical C Unit LRH, with carcinoma of breast during the above mentioned period. Name, age, sex, other relevant data, history and examination findings and results of histopathology and other investigations were recorded. RESULTS: Total of 46 patients was included in the study, out of which there were 46 female and 1 male patients. Most common age group was 40-49 years with 14 patients, followed by 50-59 years with 12 patients. Most common type of carcinoma was infiltrating ductal carcinoma with no specific features with 38 patients. Other types included 2 infiltrating ductal carcinomas of papillary type, 1 mucinous type and 1 medullary type; 3 invasive lobular carcinomas, and 1 mixed lobular and ductal carcinoma. The disease was left sided in 24 cases, right sided in 20 cases while it was bilateral in 2 cases. Upper outer quadrant of the breast was most commonly involved (n = 26). There were 2 cases of stage I, 16 stage II, 20 stage III and 08 cases of stage IV disease. There were 2 cases of grade I, 16 grade II, and 28 cases of grade III. CONCLUSION: Carcinoma breast is still a common problem presenting at a young to middle age group with invasive ductal carcinoma being the commonest variant with a high grade and a late stage of presentation due to lack of screening and awareness programs. PMID- 19999197 TI - Comparison of diagnostic efficacy of rapid diagnostic devices for dengue virus infection--a piolt study. AB - BACKGROUND: To compare the diagnostic efficacy of commercially available rapid diagnostic test devices for Dengue serology. To find out the sensitivity and specificity of rapid diagnostic devices with Elisa results as the Gold standard. METHODS: During the dengue virus epidemic in Lahore a pilot study was conducted in order to evaluate the diagnostic efficacy of two most frequently used immunochromatographic rapid test devices in public sector hospitals. The results of both the kits were compared to each other. Sensitivity and specificity was calculated against results of ELISA as the reference gold standard. RESULTS: Results of kit-A revealed a very high false negative rate when compared to ELISA where actual prevalence rate shown by ELISA was 96.0% compared to prevalence rate of 44.0% with rapid diagnostic test device kit-A. Similarly the results of rapid test device Kit-B showed high false negative results for dengue virus prevalence. Actual prevalence rate of dengue fever shown by ELISA was 96.0% where as it turned out to be 50% with the kit-B rapid test device. Comparison of Results of two kits revealed no significant difference of test positivity rates. CONCLUSION: Rapid test devices based on immunochromatographic method supplied in the public sector hospitals are not reliable diagnostic tools for screening for dengue virus infection Health authorities need to review their strategy for supply of more reliable tools during epidemics in order to avoid false negative results. PMID- 19999198 TI - Early outcome of Lichtenstein technique of tension-free open mesh repair for inguinal hernia. AB - BACKGROUND: To evaluate the early outcome of Lichtenstein's technique for repair of inguinal hernia using polypropylene mesh. METHODS: This was a descriptive study conducted over a period of twelve months from 1st July 2007 to 30th June 2008 in Surgical 'B' unit, Lady Reading Hospital, Peshawar. One hundred and twelve patients were received through the out patient department with diagnosis of inguinal hernia. Inclusion criteria was patients above the age of 18 years, reducible hernia, evidence of swelling in groin > 2 months. Exclusion criteria was age less than 18 years, chronic constipation, chronic cough, symptoms of prostatism, irreducible hernia, obstructed hernia, strangulated hernia and patients with diabetes mellitus. All the patients were subjected to inguinal mesh repair using the Lichtenstein technique with polypropylene mesh. RESULTS: Mean age of patients was 48.78 +/- 14.41 years. Sixty patients (53.6%) had right sided inguinal hernia while 46 patients (41.1%) had a left sided hernia and 6 patients (5.4%) had bilateral hernia. Sixty two patients (55.4%) had indirect hernia and 43 (38.4%) cases had direct hernia. Sixteen cases (14.3%) had previous history of surgery for hernia on the same side (recurrent hernia). Mild pain was observed in 53 cases (47.3%), moderate pain in 42 cases (37.5%), and severe pain in 17 cases (15.2%). Four patients (3.6%) in all developed a seroma Two patients (1.8%) developed a haematoma that required drainage. Three patients (2.7%) had a prolonged recovery and presented with abdominal distension. Five cases presented with infected wounds. CONCLUSION: Lichtenstein's technique of inguinal mesh repair is a safe and effective procedure but emerging trends anticipates the implementation of day case surgery. PMID- 19999199 TI - Changing spectrum of gallstone disease: an experience of 23 cases less than 10 years of age. AB - BACKGROUND: An overall increase in the incidence of paediatric cholelithiasis forms the basis of this study, which aims to investigate the overall changing clinical pattern of cholelithiasis. METHODS: This is a retrospective observational descriptive study including twenty three (23) patients with gallstones admitted and operated during June 2006-June 2008 in surgical department of a teaching hospital. All the patients with sonological evidence of gallstones, less than 10 years of age with history of acute or chronic abdominal symptoms are included in the study population. After admission all the subjects were investigated and finally operated by open approach (21 patients) during the same admission. The details of all the patients were recorded on a proforma and statistical analysis done on SPSS version 12. RESULTS: Of the total study population, there were 19 (82.6%) males and 4 (17.39%) females with a mean age of 7 years and a range of 4-10 years. Ultrasound revealed gallstones in all the patients with a varying proportion of the walls of gallbladder. The commonest presentation was abdominal pain in the right upper quadrant, which was vague, and of mild to moderate intensity. In 21 (91.30%) patients, no specific underlying cause was found while two patients (8.6%) had haematological disorder as underlying cause for the gallstones. CONCLUSION: This study indicates an alarming increase in the incidence of idiopathic gallstones in children less than 10 years of age with a distinct male predominance. PMID- 19999200 TI - Frequency, pattern and etiology of nosocomial infection in intensive care unit: an experience at a tertiary care hospital. AB - BACKGROUND: Nosocomial infection is defined as an infection which develops 48 hours after hospital admission or within 48 hours after being discharged. The objectives were to assess the frequency of nosocomial infection in patients admitted to intensive care unit (ICU) and to determine the etiological factors in such patients. It was an Observational Study and conducted in Intensive Care Unit, Liaquat University Hospital Hyderabad Sindh Pakistan from January 2008 to November 2008. METHODS: All patients above 16 years of age admitted in the ICU for more than 48 hours and developed clinical evidence of infection that did not originate from patients' original diagnosis at the time of admission, were included in the study. Data was entered in a proforma and analyzed using SPSS version 10.0. RESULTS: During the study period, 97 out of 333 patients acquired nosocomial infection. The frequency of nosocomial infection was 29.13%. Respiratory tract infection was seen in 29 (30.1%), urinary tract infection in 38 (39.1%) and blood stream infection in 23 (23.7%) patients. Other infections we identified were skin, soft tissue, wound and gastrointestinal tract infections. CONCLUSION: Patients admitted in intensive care unit are at more risk of acquiring nosocomial infection from different sources. It is suggested that proper nursing care, sterilization and disinfection of instruments and equipment and careful handling of invasive procedures are the best tool to control these life threatening infections. PMID- 19999201 TI - Pattern of change in the frequency of Helicobacter pylori with perforated duodenal ulcer. AB - BACKGROUND: Peptic ulcers were believed to be caused by stress, dietary factors, and gastric acid, but the link between H. pylori and peptic ulcers was identified in 1983. To see the frequency of Helicobacter pylori infection in patients with perforated duodenal ulcer and advise eradication therapy in these patients. This cross sectional study was conducted in Surgical Unit Hayatabad Medical Complex, Peshawar, during January 2007-June 2008. METHODS: A total of 50 cases were included in the study. All cases presenting to our unit with acute perforated duodenal ulcer were recruited. After resuscitation and baseline investigations, all underwent emergency laparotomy via upper midline incision, after thorough peritoneal lavage, the perforation margins were freshened and closed over an omental patch. Serum from every patient was tested for H. pylori and accordingly managed. RESULTS: Out of the 50 cases, 45 were males, and 5 were females. Age ranged from 20- 80 years old. All patients underwent emergency laparotomy. Postoperatively, all were started on PPI treatment and serum testing for H. pylori was done. Thirty-four (68%) turned out positive and 16 (32%) were found to be negative for H. pylori. CONCLUSION: There is still a high frequency of H. pylori infection in patients with perforated duodenal ulcer. But comparing these results with the various data available, there is a significant decline in H. pylori positive perforated duodenal ulcer patients. PMID- 19999202 TI - Ki-67 proliferating index and histological grade, type and stage of colorectal carcinoma. AB - BACKGROUND: The objective of this study was to assess the relationship of histological type, grade and stage of colorectal carcinoma with proliferative activity as measured by Ki-67 LI. It was a descriptive study and conducted at the Department of Pathology, University of Health Sciences, Lahore. METHODS: Ki-67 is a protein associated with cell proliferation and is expressed in all the phases of cell cycle except G0. In the present study, Ki-67 expression in 50 patients with colorectal adenocarcinomas was observed using immunohistochemistry with monoclonal antibody MIB-1. RESULTS: Ki-67 LI was high in well and moderately differentiated adenocarcinomas (mean Ki-67 LI 392.50 +/- 56.58 and 342.24 +/- 96.84 respectively) as compared to poorly differentiated adenocarcinomas (mean Ki 67 LI 250.00 +/- 113.46). Ki-67 LI was high in non mucinous adenocarcinomas than mucin secreting and signet ring cell adenocarcinomas (mean Ki-67 393.93 +/- 55.91 vs 220.00 +/- 49.72 and 200.00 +/- 79.05 respectively). As regard the Dukes' staging, Ki-67 LI was high in colorectal carcinomas in Dukes' stage B than tumours in Dukes' stage C (Mean Ki-67 LI 360.13 +/- 90.03 vs 241.66 +/- 101.31 respectively). CONCLUSION: The proliferative activity as measured by Ki-67 antibody is related to histological type, grade and stage. PMID- 19999203 TI - Penile fracture: experience at Ayub Teaching Hospital. AB - BACKGROUND: Penile fracture is a relatively rare traumatic rupture of the tunica albuginea of one or both corpora cavernosa of an erect penis. It is a real urological emergency which needs early assessment and surgical management. METHODS: Twelve (12) cases of penile fracture were reviewed from July, 1997 to July, 2007 in the Department of Urology, Ayub Teaching Hospital Abbottabad. All cases presented with classical history of penile fracture and the diagnosis was made on the basis of history and clinical examination only. RESULTS: All the patients underwent immediate surgical repair with well preserved potency and excellent overall results. CONCLUSION: Penile fracture has typical signs. Standard treatment consists of immediate surgical repair of penile fracture with a low incidence of late complications. Post op complications including urethral strictures and erectile dysfunction should be ruled out by regular follow-up. PMID- 19999204 TI - Evaluation of tooth size discrepancy in different malocclusion groups. AB - BACKGROUND: Orthodontic diagnosis and treatment planning requires the patient's thorough history, extra and intra oral examination, analysis of diagnostic records comprising of orthodontic photographs, necessary radiographs and properly trimmed study casts. Tooth-size is the sum of mesio distal widths of the maxillary and mandibular teeth. For ideal occlusion, teeth in both arches should be proportional in size. If larger teeth in one jaw are occluded with smaller teeth in the other jaw, ideal occlusion is not achieved. Tooth size analysis was presented by Bolton in 1958. The ratio for anterior segment was derived to be 77.2 +/- 0.22% and 91.3 +/- 0.26% for the whole arch. METHODS: A quantitative study was carried out at the Armed Forces Institute of Dentistry (AFID), Rawalpindi, Pakistan, a tertiary care facility. Cross sectional data was gathered from the study casts of patients undergoing orthodontic treatment at AFID, after obtaining informed written consent from them. 135 out of 200 study casts were filtered based upon the inclusion and exclusion criteria. ANB angle and witt's appraisal were traced on corresponding lateral cephalograms. The mesio distal widths of all maxillary and mandibular teeth from right first molar to left first molar were calibrated with the use of a manual caliper. The readings were then used to compute the anterior and total Bolton ratios. RESULTS: Significantly higher mean anterior tooth ratios were found for Class II (p < 0.01) patients. All other ratios were within close range of Bolton's norms. CONCLUSIONS: Skeletal Class II patients showed a tendency towards higher mesiodistal widths of teeth in the mandibular anterior region. PMID- 19999205 TI - Bone stress injuries in the Army cadets of Pakistan Military Academy. AB - BACKGROUND: Bone stress injuries are common among army cadets during training period and these injuries give rise to morbidity and loss of training hours. Some cadets with stress fractures re-join their duties while some are kept out of services resulting in attrition and economic loss. A retrospective analysis of bone stress injuries of one hundred and twenty cadets between June 2006 and Dec 2007 was conducted from the case documents available at Institute of Nuclear Medicine, Oncology & Radiotherapy (INOR) Abbottabad. RESULTS: The occurrence of stress fractures among cadets at Pakistan Military Academy, presenting with exercise induced pain was 68.3% amongst these cadets. Seventy fours percent of the fractures were labelled grade-I. High grade fractures were confined to tibia. Tibia was the commonest site of stress fractures in both sexes, accounting for about 73% of all stress fractures. Fifty percent of these patients were labelled as having shin splints and 3% as thigh splints. CONCLUSIONS: Most of the stress injuries were confined to the tibia. The referral was considered early as most of the stress injuries were of low grade. PMID- 19999206 TI - Role of barium meal examination in diagnosis of peptic ulcer. AB - BACKGROUND: Peptic ulcer is a sore in the protective lining (mucosal lining) of the gastrointestinal tract and develops when the lining is damaged. The objectives of this Descriptive Validational study were to determine the validity of Barium Meal examination in the diagnosis of peptic ulcer disease in comparison to the gold standard, i.e., endoscopic evaluation in peptic ulcer disease. The study was conducted at Radiology Department Khyber Teaching Hospital Peshawar from November 2000 to March 2004. METHODS: A total of 115 patients with signs and symptoms of peptic ulcer disease were selected for this study and were subjected for this diagnostic modality. The diagnosis of benign/malignant peptic ulcer was made by barium meal examinations. In all these patients the diagnosis was later on confirmed by endoscopy and or surgery. RESULTS: Of the 115 patients, 80 were male and 35 were female patients. Their ages ranged from 27-75 years with mean age of 49 years. Fifty two patients had duodenal ulcer, 30 patients gastric ulcer, and 33 patients had normal radiological findings. In 6 out of 30 patients with gastric ulcer had radiological evidence of malignant gastric ulcer. CONCLUSION: Efficiency of Barium meal examination in diagnosis of peptic ulcer is good and most of peptic ulcers can be diagnosed by this method. PMID- 19999207 TI - Evaluation of the aetiological spectrum of obstructive jaundice. AB - BACKGROUND: Jaundice is a common problem in medical and surgical gastroenterological practice. The surgical jaundice can be caused by the obstruction of the bile duct as with gall stones, strictures, malignancy, such as cholangiocarcinoma (in which the jaundice is persistent and progressive), periampullary carcinoma, carcinoma gall bladder and carcinoma head of pancreas. The objective of this descriptive study was to evaluate the Etiological spectrum of obstructive jaundice. METHODS: A prospective, descriptive study was carried out at Surgical Unit-II Holy family Hospital, Rawalpindi, from mid of May 2006 till March 2007. Sixty patients, who presented in the surgical OPD of Holy family Hospital, were included in the study. Thorough history and physical examination was followed by biochemical tests and various investigations like USG abdomen, ERCP, CT-Scan, & MRCP and histopathology. The data was analyzed using SPSS ver 14.0. RESULTS: Of the 60 patients; 40 (66.66%) were male and 20 (33.33%) were female, their mean age being 49.50 years. Malignant obstructive jaundice was seen in 34 (56.66%) patients while 26 (43.33%) had benign etiology. Amongst the commonest symptom; clay coloured stools (75%) was more frequent in patients with malignant disease whereas abdominal pain (51.66%) was most common in benign conditions. Commonest malignancy was Carcinoma (Ca) of the head of pancreas 18/60 (30%) followed by Ca gall bladder 8/60 (13.33%), cholangiocarcinoma 7/60 (11.66%), and periampullary carcinoma 1/60 (1.66%). Choledocholithiasis 21/60 (35%) was the commonest benign cause followed by stricture of common bile duct 3/60 (5%) and acute pancreatitis 2/60 (3.33%). CONCLUSION: Obstructive jaundice is common amongst females and the cause is mostly malignant. Ca head of pancreas is the commonest malignancy while Choledocholithiasis is the commonest benign cause. USG, ERCP and CT-Scan are important diagnostic modalities for evaluation of patient with obstructive jaundice with ERCP having the additional advantage of being therapeutic as well. PMID- 19999208 TI - Rapid isolation of Mycobacteria--need of the hour in our settings. AB - BACKGROUND: To compare Bactec MGIT 960 with LJ media in terms of time taken for the initial isolation of mycobacteria. METHODS: A total of 100 AFB (acid fast bacillus) positive sputum samples were processed and inoculated in both the Lowenstein Jensen (LJ) media and mycobacterium growth indicator tube (MGIT) tubes. RESULTS: Of the 100 samples, positive growth was obtained from all the samples on both the MGIT and LJ media. In MGIT 53% samples grew in 4 days, 30% in 5 days and 17% in 6 days (Mean = 4.6 days) while on LJ media, 44% grew in 30 days, 20% in 35 days and 36% in 44 days (Mean = 37 days). Significant difference was observed between two systems with a p-value of less than 0.05. CONCLUSION: Bactec MGIT 960 is a much faster and efficient system for the initial isolation of mycobacteria. PMID- 19999209 TI - Type 2 diabetes mellitus: how well controlled in our patients? AB - BACKGROUND: Type 2 Diabetes mellitus has reached epidemic proportions worldwide and Pakistan is no exception. This study was done to see the glycaemic control of our diabetic patients by estimating Glycosylated haemoglobin & Fasting blood glucose as poor control leads to significant complications causing enormous human suffering & socioeconomic burden. METHOD: This Cross-sectional study was conducted on Type 2 diabetic patients coming to medical OPD and medical B ward of Ayub Teaching Hospital between March-September 2007 fulfilling the inclusion criteria. RESULTS: Among 100 patients with type 2 diabetes forty two had HbA1c more than 7.5%, while seventy had fasting blood glucose more than 120 mg/dl. All patients with HbA1c more than 7.5% had increased fasting blood glucose. While thirty out of seventy patients with fasting blood glucose more than 120 mg/dl had HbA1c less than 7.5%. None of the patients with fasting blood glucose less than 120 mg/dl had HbA1c more than 7.5%. CONCLUSIONS: Significant number of patients (42%) had poor control of diabetes as revealed by HbA1c, with FBG showing poor control in even more patients, i.e., 70%. However their blood glucose estimation was not frequent enough as required. Blood glucose results can be spuriously high and may lead to frequent change/increase in the dose of hypoglycaemic medications. This can lead to poor compliance as well as psychological trauma to patients. HbA1c on the other hand is easy to interpret, reflects long term glycaemic control and cost effective. We recommend its more frequent use along with blood glucose for better glycaemic control and decreased chances of complications. PMID- 19999210 TI - Anterior decompression, fusion and plating in cervical spine injury: our early experience. AB - OBJECTIVE: To evaluate the efficacy of anterior cervical decompression, fusion and titanium plate fixation in sub axial cervical spine injuries in respect of neurological outcome, postoperative stability and early rehabilitation. The Descriptive case series study was conducted at Department of Neurosurgery, Liaquat University Hospital, Jamshoro, Sindh Pakistan during year 2005 to 2007. METHODS: Patients with cervical spine injuries were admitted during study period were included in this study. All cases were evaluated for their clinical features. During initial phase, level and degree of neurological injury was assessed using ASIA impairment scale. Cervical traction was applied to all patients. Operative and post operative record with x-rays and MRI were maintained. Patients with Injury to C3-6 underwent decompression, fusion and local titanium plate implant fixation by anterior approach. The follow-up ranged from 6 to 12 months with clinical and radiological assessment. RESULTS: 37 cases of sub axial cervical spine injuries included in this study during year 2005 to 2007. Out of these, 28 (75.67%) were males and 9 (24.32%) females. Age range was 8-60 years mean (32-40%). Common mode of injury was fall. Post operative follow up showed good clinical and radiological outcome, bony fusion and favour early rehabilitation. No immediate complication found except temporary dysphagia. CONCLUSION: Anterior decompression, fusion and titanium plate fixation is an effective method with good neurological and radiological outcome. PMID- 19999211 TI - Preventive role of zinc chloride against toxicity of ciprofloxacin on the growing cartilage of Wistar albino rat litter. AB - OBJECTIVE: To assess the preventive role of zinc chloride on toxicity of ciprofloxacin administration in Wistar albino rat litter. It was a Prospective experimental study. The study was carried out in the Department of Anatomy, Basic Medical Sciences Institute, Jinnah Postgraduate Medical Centre, Karachi, Pakistan during March 2002 to February 2003 one year study. METHOD: Ciprofloxacin and zinc chloride were administered to newly born albino rat litters separately and simultaneously at a dose of 20 mg/kg body weight and 1200 microg/Kg body weight respectively, intraperitonealy twice daily from 1 - 14 day after birth. The animals were sacrificed by deep ether anaesthesia. The fore and hind limbs were dis-articulated from the axial skeleton, soft tissue was removed and bones were fixed in 10% buffered formalin. Decalcification was done in 10% nitric acid and 10% formic acid changes. After paraplast embedding, 4 microm thick longitudinal sections of proximal & distal ends of long bones were cut by a rotary microtome. Routine staining with haemotoxylin and eosin was performed. Histomorphometery was done to measure the thickness of epiphyseal cartilage and was compared with similar values of the control animals. The results were statistically analyzed to evaluate the significance. RESULT: Our study revealed that ciprofloxacin administration in new born albino rat litter decreased the width of epiphyseal growth plate cartilage by 13.7 +/- 0.42 microm, 10.43% in humerus and 6.6 +/- 1.2 microm 4.72% in femur as compared to control, whereas, simultaneous zinc chloride administration restricted the decrease to 1.27 microm +/- SD in humerus and 2.05 microm +/- SD in femur. CONCLUSION: Simultanous zinc chloride administration minimized the epiphseal cartilage damage induced by ciprofloxacin in Wistar albino rat litter. PMID- 19999212 TI - Correlation of severity of ST segment elevation in acute inferior wall myocardial infarction with the proximity of right coronary artery disease. AB - BACKGROUND: A number of researchers have used different electrocardiographical criteria to predict the culprit vessel in acute interior wall myocardial infarction (MI) cases. Therefore, the determination of infarct related artery in AMI is extremely important with regard to prediction of potential complications, furthermole, predicting the probable site of occlusion within RCA is worthwhile because proximal occlusions are likely to cause greater myocardial damage and an early invasive strategy may be planned in such cases. Our study aimed at evaluating the ECG criteria to predict the proximity of lesion in the right coronary artery (RCA) in acute inferior wall MI cases. The Objectives were to predict the presence of a proximal lesion in right coronary artery by severity of ST segment elevation in inferior ECG leads. This cross sectional study carried out at the department of cardiology and cardiac catheterization at Jiinnah Hospital, Lahore from April 2008 to September 2008. METHODS: A total of 60 patients who suffered from inferior wall MI were included in the study who underwent coronary angiography in the first week. The ECGs of these patients were then compared with the angiographic findings to correlate the proximity of culprit lesion in RCA with the degree of ST segment elevation in inferior limb leads. RESULTS: Out of 60 patients, 29 (48.4%) had the culprit lesion in proximal, 23 (38.5%) in mid and 8 (13.4%) in distal RCA. Patients with proximal RCA disease showed a mean ST segment elevation of 12.55 +/- 1.38 mm, with mid RCA disease 8.39 +/- 0.89 mm and with distal RCA disease 6.0 +/- 0.54 mm. CONCLUSION: This study demonstrated that the severity of ST segment elevation was correlated with proximity of RCA lesion. PMID- 19999213 TI - Urbach-Wiethe disease: experience at a tertiary care hospital in Abbottabad, Pakistan. AB - BACKGROUND: Urbache-Wiethe disease (Lipoid Proteinosis) is a rare autosomal recessive disorder characterized by the deposition of an eosinophilic hyaline like material in the skin, larynx, mucous membranes, brain, and other internal organs. METHODS: A survey of one year duration was carried out prospectively at the Department of Dermatology, Ayub Teaching Hospital Abbottabad to document cases of lipoid proteinosis. Cases were selected from the outpatients department on the basis of clinical presentation and were subjected to detailed examination and investigations after admission. RESULTS: Five cases were diagnosed as suffering from Lipoid Proteinosis over the study period. All had typical features of hoarseness, skin lesions and tongue involvement. All were born of consanguineous parents. Three (60.0%) cases also gave a history of involvement of other family members, particularly cousins. CONCLUSION: This rare disease occurs in Hazara Division of North West Frontier Province of Pakistan with an as yet undetermined frequency and clinical suspicion is warranted to diagnose cases with the typical presentation. PMID- 19999214 TI - Inherited dyslipidaemic disorders contributing to coronary heart disease. AB - BACKGROUND: Lipoprotein (a) [Lp (a)] is an established independent risk factor for premature myocardial infarction (MI)/coronary artery disease (CAD). The study was conducted to determine the value of Lp (a) in prediction of CAD or MI in the offspring at risk. METHODS: A total of 160 subjects were investigated. Serum Lp (a) was measured by ELISA, serum total cholesterol, triglycerides (TG) and HDL Cholesterol by enzymatic colorimetric methods using standard kits. RESULTS: Differences in levels of total Lp (a) and cholesterol were observed between patients and controls. Both Lp (a) (16.23 +/-1.95 mg/dL) and cholesterol (175.00 +/- 7.60 mg/dL) of group A (patients) were higher than the corresponding controls. However an opposite trend in results was noted for serum HDL Cholesterol in patients vs. controls. CONCLUSION: Persons found to have elevated levels of Lp (a) should focus on controlling the known modifiable risk factor for heart disease, especially smoking, hypercholesterolemia, obesity, hypertension and sedentary life style. PMID- 19999215 TI - Percutaneous needle peritoneal biopsy in the diagnosis of exudative ascites. AB - BACKGROUND: Percutaneous needle peritoneal biopsy in diagnosis of exudative ascites has gained wide acceptance and many workers have utilized it with a high diagnostic yield and no significant complications. Present study has been carried out to determine the efficacy of percutaneous needle peritoneal biopsy in the diagnosis of exudative ascites of unknown aetiology. METHODS: It is a descriptive case study conducted in Medical 'C' Unit, Lady Reading Hospital, Postgraduate Medical Institute, Khyber Medical University Peshawar over a period of 2 years, i.e., from Nov, 2003 to December 2005. A total of 45 patients having unexplained exudative ascites underwent blind needle peritoneal biopsy. The biopsy specimen was subjected to histopathology. Ascitic fluid was also obtained for analysis. Post biopsy patients were observed for 24 hours for any untoward complications. RESULTS: A total of 45 patients (17 male and 28 female) with age range from 20 to 65 years and having exudative ascites were studied. The commonest presentation of our patients was abdominal distension (93.3%), pain abdomen (46.67%), fever (44.4%) and weight loss (33.3%). Histopathology of the peritoneal biopsies was reported as follows. Eighteen cases (40%) showed non specific chronic inflammation, 10 (22.2%) cases showed caseating granulomatous inflammation suggestive of tuberculosis and 6 (13.3%) cases showed metastatic adenocarcinoma. In one patient peritoneal mesothelioma was reported. In the remaining 10 patients (22.2%) biopsies were either non representative or inconclusive. The ascitic fluid showed predominantly lymphocytes in 86.6% of cases. Only three patients were reported to be having atypical cells on fluid cytology. The procedure was found safe. No patient was lost due to complications related to the procedure. Only one patient had evidence of intra peritoneal bleed. The commonest problem post biopsy was pain (91.1%) and mild swelling (53.3%) at biopsy site. CONCLUSION: Peritoneal biopsy is fairly safe and inexpensive procedure with good diagnostic efficacy in patients with undiagnosed exudative ascites. PMID- 19999216 TI - Histological patteren of nephrotic syndrome in elderly patients. AB - BACKGROUND: Renal biopsy is a safe and informative technique that has played a key role in the evolution of nephrology as a specialty. This cross-sectional descriptive study was aimed to study the histological pattern of nephrotic syndrome in elderly patients. METHODS: This study was conducted from January 1998 to December 2005 (8 Years) in the Department of Nephrology, Govt. Lady Reading Hospital, Peshawar, Pakistan. We performed renal biopsy in 153 patients aged 60 years and above, admitted with nephrotic syndrome-proteinuria with protein > 3 gm/24 hours with or without oedema, hypoalbuminemia, hyperlipidemia, acute nephritic syndrome-red blood cell casts or dysmorphic red cells on urine microscopy, temporally associated with acute renal failure with or without new onset hypertension, acute renal failure-sudden decline in renal functions developing in days to weeks causing retention of nitrogenous wastes and rapidly progressive renal failure-sub acute decline in renal functions developing over weeks to months. RESULTS: Significant differences in the histological pattern are noticed when compared with the studies of the other centres. Minimal Change Disease (MCD) was the commonest histological pattern (40%), followed by Membranous Nephropathy (21%), Crescentic Nephritis (19.6%), Focal Segmental Glomerulosclerosis (FSGS) (6.5%), Hypertensive Nephropathy (5.8%), Messangiocapillary Glomerulonephritis (MCGN) (3.9%) and Diffuse Proliferative Glomerulonephritis (DPGN) (3.2%). PMID- 19999217 TI - Presentation, staging and diagnosis of lymphoma: a clinical perspective. AB - BACKGROUND: Due to lack of awareness among health professionals, lymphoma is often misdiagnosed. This study was done to evaluate the clinical features and histopathologic subtypes of lymphoma. METHODS: Sixty diagnosed cases of lymphoma were selected (aged 12-65 years) from medical units of Civil Hospital Karachi, during 1993 to 1998. Clinical history, physical examination and basic laboratory investigations including imaging procedures were done in all the patients. The diagnosis of lymphoma was based on histology, following the International Working Formulation classification system. This included lymph node biopsy and in some cases, biopsy of the bone marrow. The Ann Arbor Staging Classification was used to classify the extent of disease. RESULTS: Out of 60 cases of lymphoma, 81.6% (49 cases) were diagnosed as non-Hodgkin's lymphoma and 18.3% (11 cases) as Hodgkin's disease, with an overall male predominance. Both categories exhibited a bimodal age distribution. Lymphadenopathy was the commonest presenting features in both the types of lymphomas; however, patients with Hodgkin's disease had a prominence of 'B' symptoms, whereas abdominal signs and symptoms were more common in non-Hodgkin's lymphoma. On histopathology, majority of non-Hodgkin's lymphomas (91.8%) showed a diffuse pattern, while mixed cellularity was the commonest type seen in Hodgkin's disease (81.8%). CONCLUSION: Non-Hodgkin's lymphoma was 4 times more common than Hodgkin's disease. The vast clinical spectrum of lymphoma sometimes delays its diagnosis, leading to its eventual presentation in late stages. A general awareness is hence required among the health professionals regarding its varied clinical presentations. PMID- 19999218 TI - Surgical outcome of transpedicular fixation in thoracolumber fractures. AB - BACKGROUND: Development of transpedicular screw fixation techniques and instrumentation systems has brought short-segment instrumentation (fixation of one normal vertebra above and below an injured segment) into general clinical practice. The purpose of this study is to report the surgical outcome of thoracolumber fractures treated with short-segment pedicle instrumentation. METHODS: A retrospective review of all surgically managed thoracolumbar fractures during two years was performed. The 84 surgically managed patients were instrumented by the short-segment technique. Patients charts, operation notes, preoperative and postoperative radiographs, computed tomographic scans, magnetic resonance imaging (MRI) was done. Neurological findings (Frankel functional classification), and follow-up records up to 6 months were reviewed. RESULTS: Transpedicular fixation was performed in 84 cases including 52 male and 32 female with male to female ratio 1.6:1. Mean +/- SD of age was 40 +/- 13.75 years (range 15-60). The level of injuries was different in different age groups. Outcome was assessed on Frankle grading. No patient showed an increase in neurological deficit. Most of the patients showed improvement to the next grade. Screw breakage occurred in 8 cases, bed sores in 16 cases and deep vein thrombosis in 3 cases. Misplaced screw in 5 cases. Eight cases got wound infection. CONCLUSION: Although long- term follow-up evaluation needs to verified, the short-term follow up results suggest a favourable outcome for short-segment instrumentation. PMID- 19999219 TI - Tube thorocostomy: management and outcome in patients with penetrating chest trauma. AB - BACKGROUND: Penetrating chest trauma is common in this part of the world due to present situation in tribal areas. The first line of management after resuscitation in these patients is tube thoracostomy combined with analgesia and incentive spirometry. After tube thoracostomy following surgery or trauma there are two schools of thought one favours application of continuous low pressure suction to the chest tubes beyond the water seal while other are against it. We studied the application of continuous low pressure suction in patients with penetrating chest trauma. This Randomized clinical controlled trial was conducted in the department of thoracic surgery Post Graduate Medical Institute Lady Reading Hospital Peshawar from July 2007 to March 2008. The objectives of study were to evaluate the effectiveness of continuous low pressure suction in patients with penetrating chest trauma for evacuation of blood, expansion of lung and prevention of clotted Haemothorax. METHODS: One hundred patients who underwent tube thoracostomy after penetrating chest trauma from fire arm injury or stab wounds were included in the study. Patients with multiple trauma, blunt chest trauma and those intubated for any pulmonary or pleural disease were excluded from the study. After resuscitation, detailed examination and necessary investigations patients were randomized to two groups. Group I included patients who had continuous low pressure suction applied to their chest drains. Group II included those patients whose chest drains were placed on water seal only. Lung expansion development of pneumothorax or clotted Haemothorax, time to removal of chest drain and hospital stay was noted in each group. RESULTS: There were fifty patients in each group. The two groups were not significantly different from each other regarding age, sex, pre-intubation haemoglobin and pre intubation nutritional status. Full lung expansion was achieved in forty six (92%) patients in group I and thirty seven (74%) in group II. Partial lung expansion or pneumothorax was present in three (60%) in group I and 10 (20%) in group II. One patient in group I and three (6%) patients in group II had no response. The mean time to removal of chest drains were 8.2 +/- 3.14 days in group I and 12.6 +/- 4.20 days in group II. The length of hospital stay was 7.2 +/- 2.07 days and 12.4 +/- 3.63 days in group I and II respectively. Clotted Haemothorax requiring surgery developed in three (6%) patients in group I and 8 (16%) patients in group II. CONCLUSION: Placing chest tubes on continuous low pressure suction after penetrating chest trauma helps evacuation of blood, expansion of lung and prevents the development of clotted Haemothorax. It also reduces the time to removal of chest drains, the hospital stay and the chances of surgery for clotted Haemothorax or Empyema. PMID- 19999220 TI - Impact of anthropmetric measures on sural nerve conduction in healthy subjects. AB - BACKGROUND: Nerve conduction studies are an invaluable aid to investigate and quantify the physiological activity of peripheral nerves. These include measurement of sensory and motor conduction velocities and latencies of peripheral nerves. These nerve conduction parameters may be affected by anthropometric factors like age, sex, height, weight and BMI. In this observational study the impact of these anthropometric parameters was measured on sensory nerve conduction velocity (SNCV) and sensory latency (SNSL) of sural nerve. METHOD: Sural nerve conduction studies were conducted in EMG room of Neurology section of Medical Unit 1 at Sir Ganga Ram Hospital, Lahore on Electromyograph by Nihon Kohden MEB-5304K. RESULTS: After following strict exclusion criteria by careful history and physical examination 25 normal healthy subjects recruited from local population by random sampling technique. The age range was 40-70 years with mean age 47.2 +/- 9.23 years. Gender distribution was 60% females and 40% males. The mean SNCV was found to be 53.16 +/- 5.68 meter per second (m/sec) and mean SNSL was 2.81 +/- 0.45 milliseconds (msec). CONCLUSION: It was found that the mean sural nerve conduction velocity was higher and the mean sensory latency was shorter in healthy subjects than that of the Western races. Moreover, the SNCV decreases and SNSL increases significantly with age. PMID- 19999221 TI - Abdominal tuberculosis an experience at Ayub Teaching Hospital Abbottabad. AB - BACKGROUND: Abdominal tuberculosis is one of the common diseases in our country. This study was performed at Surgical A Unit Ayub Teaching Hospital Abbottabad from August 2006 to December 2007 to asses the clinical presentation of abdominal tuberculosis and its management. METHODS: All patients presenting to outpatient department with clinical feature suggestive of abdominal tuberculosis were included in the study. They were investigated. On the basis of clinical presentation, patients were divided in two groups. Patients with acute abdomen (peritonitis, intestinal obstruction) were prepared for laparotomy and operated upon. Required surgical procedure performed and tissue diagnosis was obtained. Patients with sub-acute obstruction, chronic pain abdomen and mass abdomen with out peritonism were managed conservatively. These patients were started on anti TB drugs on the basis of clinical and laboratory assessment. Empiric therapeutic trial was conducted for at least for 3 months with standard four drugs regimen. They were sent home on 9 month course of Anti TB drugs and were advised to come for follow up twice a month. On reassessment good clinical response was considered abdominal tuberculosis and anti TB continued with monthly follow up. In case of no response patients were operated. Required surgical intervention performed and tissue was taken to establish diagnosis. Detailed history, family history, examination findings, results of investigations, operative findings of the histologically proven cases of abdominal tuberculosis were recorded on a separate proforma and analyzed. RESULTS: Amongst 76 patients majority were females 52 (68.40%). Most of the patients were young with mean age of 34 years. Abdominal pain was the most common presentation 73 (96%) followed by fever and anorexia. Tender lower abdomen as found in 53 (70%) patients and mass abdomen was found in 35 (46%). Family history of TB was present in 20 (26%). Fifty three (70%) patients underwent laparotomy. Bands and adhesion was the most frequent finding on laparotomy. CONCLUSION: Abdominal TB is more common in female and abdominal pain fever and nausea are the most common presentations. PMID- 19999222 TI - Body mass status of school children of Dera Ismail Khan, Pakistan. AB - BACKGROUND: Childhood obesity is a global epidemic involving both developed and developing countries. It is a stare of over-nutrition with long-term complications such as dyslipidemia, hypertension, and coronary artery disease and type-2 diabetes. Underweight is the result of under nutrition leading to reduction in growth and development of every body organ especially the Central Nervous System. Long-term under-nutrition causes failure in linear growth (height) of the child. Growth is further retarded by the repeated attacks of respiratory infections, diarrhea and anemia as a result of reduced immunity. METHODS: This study was carried, out eight primary schools of Dera Ismail Khan (Private, semi government organizations, and welfare foundations) having mixed population with some of the wards belonging to high socioeconomic group. Thorough clinical examination excluded those suffering from chronic heath problems. Height and weight of each one was taken body mass index of determined according to 'Quatelet's' index. Body mass index number was plotted on the CDC S age and gender specific growth charts 2-20 years for BMI-for age percentile and body mass status (underweight, normal weight, overweight/at the risk of overweight and obese/overweight. RESULTS: Total 1338 school going children (6-11 years) were examined with 865 (67.75%) boys and 471 (35.25%) as girls. 13.39%, 72.15%, 8.83% and 5.61% as underweight, normal weight, overweight and obese respectively. Percentage of underweight was higher in girls (25%) than boys (13.22). Percentage of obesity was higher (5.17%) in boys than girls (1.39%). CONCLUSION: Awareness about balanced diet, improvement in the level of education and socioeconomic conditions, easy access to health facilities and prevention of the gender discrimination, are the remedial measures to be taken to redress the situation. PMID- 19999223 TI - Evaluation of diagnostic accuracy of APRI for prediction of fibrosis in hepatitis C patients. AB - BACKGROUND: Several non-invasive markers are being used to assess the structural liver damage in patients with chronic hepatitis C (CHC). We evaluated Aspartate aminotransferase (AST) to platelet ratio index (APRI) in comparison with Metavir scoring for assessing the severity of hepatic fibrosis in the CHC patients in district Rawalpindi. METHODS: One hundred twenty CHC patients, naive for HCV treatment, underwent liver biopsy in tertiary care hospitals of district Rawalpindi, participated in the study. Liver biopsies were reviewed by Metavir scoring system. Serum AST was analyzed by IFCC method. Platelets were measured on a haematology Analyzer. Patients with mild fibrosis (F0, F1) were differentiated from significant fibrosis (F2, F3, F4) and those with mild/moderate fibrosis (F0, F1, F2) from advanced fibrosis (F3, F4) based on APRI score as compared to liver biopsy. RESULTS: Liver biopsies examination revealed that out of 120 patients 10 (8.3%) had no fibrosis (F0), 46 (38%) portal fibrosis (F1), 34 (28%) septal fibrosis (F2), 21 (18%) bridging fibrosis (F3) and 9 (8%) cirrhosis (F4). APRI correctly classified 58 (48%) patients of significant fibrosis with AUC = 0.82 (95% CI, 0.73-0.88) at cut-off 0.5 and 1.5 with negative predictive value (NPV), Positive predictive value (PPV), sensitivity and specificity of 78%, 72%, 66%, 83% and 58%, 90%, 41% and 90% respectively. Eighty-seven (66%) CHC patients were correctly classified for advanced fibrosis with AUC = 0.87 (95% CI 0.79-0.94) at cutoffs 0.90 and 1.75 with a 95%NPV at 0.90 and 78% PPV at 1.75. CONCLUSION: APRI could correctly identify significant fibrosis in 48% and advanced fibrosis in 66% cases with acceptable degree of diagnostic accuracy in CHC patients in our clinical practice. PMID- 19999224 TI - Frequency of ABO and Rhesus blood groups in District Swat, Pakistan. AB - BACKGROUND: Up till now about 400 red cells antigen have been identified. The majority are inherited by Mendelian Fashion. The ABO blood group system was first to be identified and RH blood group system was the 4th one, both are most important for blood transfusion purposes. This study is conducted to determine the frequency of ABO and Rhesus (Rh) blood groups in District Swat, NWFP, Pakistan. It is a cross sectional prospective study and was conducted at Saidu Teaching Hospital district Swat, over a period of one year. (1st Jan. 2007 to 31st Dec, 2007). METHODS: A total of 22897 subjects were included in this study. Patients were collected from different wards of Saidu Teaching Hospital while the donors from common population. From each subject blood was collected, ABO and Rh blood grouping were carried out by tile method using commercially prepared anti sera. The frequency of each type was calculated. RESULTS: Out of 22897 subjects 17141 (74.86%) were male subjects and 5756 (25.140%) were female. Out of 17141 male subjects 15597 (90.99%) and out of 5756 female subjects 5040 (87.56%) were found to be Rh-positive. The frequency of Rh-negative group in male subjects were (9.01%) where as in female subjects were (12.22%). The frequency of A, B, O and AB groups in Rh-positive male subjects were 25.63%, 29.54%, 26.04% and 9.78%, amongst female subjects, it was 24.53%, 28.06%, 25.54% and 9.43% respectively. In Rh-negative male subjects the frequency of A, B, O and AB is 2.25%, 2.88%, 3.01% and 0.88%, while amongst females it is 3.54%, 4.24%, 3.74% and 0.92% respectively. CONCLUSION: It is concluded from this study that frequency of Rh positive blood group is B, O, A, and AB in both gender. Where as the most common Rh-negative in male and female subjects are O, B, A, AB, and B, O, A, and AB respectively. PMID- 19999225 TI - MUC1 based breast cancer vaccines: role of post translational modifications. AB - BACKGROUND: Vaccine development is one of the most promising fields in cancer research. After autologous transplantation, due to low tumour burden, patients are more likely to respond immunologically to a cancer vaccine. MUC1 with its adhesive and antiadhesive functions, immunostimulatory and immunosuppressive activities, is therefore a good candidate for breast cancer vaccine. A structure based insight into the immunogenicity of natural MUC1 glycoforms, of its sub domains, motifs and post translational modification like glycosylation and myriostoylation may aid the design of tumour vaccines. METHODS: Primary sequences of human MUC1 were retrieved from the SWISSPROT data bank. Protein pattern search: The primary sequence of Human MUC1 was searched at PROSITE (a dictionary of protein sites and patterns) database. RESULTS: Our study observes that post translational modifications play an important role in presenting MUC1 as a candidate for breast cancer vaccine. CONCLUSION: It is found that the phosphrylation and glycosylation of important functional motifs of MUC1 may take part in the production of cytokines that may provide immunization. PMID- 19999226 TI - Brain natriuretic peptide, systolic and diastolic blood pressures. AB - BACKGROUND: Hypertension is an increasingly important medical and public health issue. Because of life time risk of hypertension Joint National Committee-7 introduced a new term "Pre-hypertension" for those with systolic blood pressure 120-139 mmHg and diastolic blood pressure 80-89 mmHg. Individuals prone to the development of hypertension often have a hyperdynamic circulation antedating the onset of hypertension by several years. Brain Natriuretic Peptide is a new promising cardiovascular risk marker due to its association with high blood pressure via its key role in salt and water homeostasis and hence blood pressure. This study was designed to find out the levels of plasma Brain Natriuretic Peptide in normotensive, pre-hypertensive and newly diagnosed hypertensive and to observe its association with increasing values of blood pressure. METHODS: This was an cross-sectional study conducted in the Department of Physiology Basic Medical Sciences Institute, Jinnah Post Graduate Medical Centre, Karachi. Study included 85 adult males, aged between 20-60 years, non- smokers, non- diabetic and having no other chronic illness. Systolic blood pressure and diastolic blood pressure values were measured and divided into three groups on the basis of normotensive to hypertensive as stated by Joint National Committee -7. Brain Natriuretic Peptide was assayed by AxSym technology. RESULTS: Brain Natriuretic Peptide developed a positive and statistically significant correlation with both systolic and diastolic blood pressures and was also found out to be significantly raised in pre-hypertensive group. CONCLUSIONS: his study concluded that Brain Natriuretic Peptide is positively related with increasing values of both variables i.e. systolic as well as diastolic blood pressures. It also concluded out Brain Natriuretic Peptide to be significantly elevated in pre-hypertensive stage which may remain increased in sustained hypertension. PMID- 19999227 TI - Evaluation of serum leptin levels during normal pregnancy and in pre-eclampsia. AB - BACKGROUND: The present study was designed to compare the serum leptin levels in normal pregnancies with those pregnancies which are complicated by pre-eclampsia and to investigate the possibility of leptin being a marker of severity of pre eclampsia. METHODS: This was a comparative cross-sectional study, conducted at the Department of Gynaecology and Obstetric, Jinnah Postgraduate Medical Centre (JPMC), Karachi. For this purpose 45 primigravidas with normal pregnancy and 45 primigravidas with pre-eclamptic pregnancy were selected who were in their third trimester All the subjects were of the same maternal age, gestational age, height and weight. Serum leptin levels were determined by immunoenzymometric assay. RESULTS: The study included 90 patients, divided into two groups. The serum leptin levels were found to be elevated in pre-eclamptic group as compared to normal pregnancy. When comparison was made on the basis of severity of pre eclampsia, the serum leptin levels were found to be increased in severe group as compared to mild group. CONCLUSION: From this study it was concluded that the levels of serum leptin were higher in pre-eclamptic group when compared to controls during the third trimester of pregnancy indicating the possible role of leptin in the pathogenesis of pre-eclampsia. Also the elevated level of leptin may help in the prediction of the disease and may serve as a marker of severity of pre-eclampsia. PMID- 19999228 TI - Awareness of women regarding hepatitis B. AB - BACKGROUND: Hepatitis B is the major health problem globally casting an enormous burden on health care system and major source of patient's misery. It is also an important cause of hepatocellular carcinoma and is likely to remain a serious health problem resulting in substantial morbidity and mortality for several decades to come. Objective of the study was to determine the level of awareness regarding hepatitis B among women attending obstetric and gynaecology clinic at Isra university Hospital. It was descriptive study and conducted in Obstetric and Gynaecology, Outpatient Department of Isra University Hospital, Hyderabad, Sindh from 2nd January to 3rd June 2008. METHODS: Total 121 women of 20-50 years were gathered by sample of convenience. Information was collected on predesigned questionnaire containing questions regarding basic knowledge of disease, its causative organism, route of transmission and its prevention. Informed consent was taken. All the data was analyzed by computer software Programme of Social Sciences (SPSS version 11.0). RESULTS: Out of 121 women, 82 (67.76%) women correctly responded that virus is a cause of hepatitis. When they were asked about the organs affected by hepatitis B infection, 91 (75.20%) correctly mentioned liver, while 30 (24.79%) gave incorrect answer. Regarding mode of transmission of HBV 41 (33.88%) mentioned infected blood transfusion, 49 (40.49%) contaminated needles, 46 (38.0%) un-sterilised instruments, 21 (17.35%) from mother to child and only 23 (19.0%) mentioned sexual intercourse. Only 51 (42.14%) women knew that vaccine is available for prevention. CONCLUSION: This study found that the knowledge about hepatitis B virus among women is inadequate and there are certain misconceptions regarding its mode of transmission through sharing room, touching etc which needs to be clarified. Majority of women don't know that HBV infection can be prevented by using condoms. PMID- 19999229 TI - Septic induced abortions. AB - BACKGROUND: Induced abortions are not infrequent in our community. Being the only tertiary level care hospital in Hazara division, Ayub Teaching Hospital frequently receives cases of induced abortions with complications attempted by inappropriately trained personnel. It was decided to study the frequency of such cases in our admitted patients, complications and their outcome in such risky situation. METHOD: A cross sectional study was conducted from January 2006 to December 2007 in Gynae-B Unit, Ayub Teaching Hospital, Abbottabad. All admitted patients in the unit in the 1st trimester of pregnancy and handling outside the hospital were included in the study. RESULTS: In two years period total Gynae admissions were 1090. There were 152 (13.95%) cases of spontaneous 1st trimester abortions and 52 (4.7%) cases of induced abortions which are included in the study. Age range of the patients was between 20-45 years, with a mean of 35 years. Majority of the patients were grand multigravidas belonging to lower socioeconomic group and not using any contraception. Twenty patients (38.4%) presented with heavy per vaginal bleeding due to incomplete nature of the procedure. Twelve patients 23% presented in shock due to excessive per vaginal bleeding. Ten patients 19.2% had uterine perforation and intraperitoneal haemorrhage. Two patients 3.8% had gut injury associated with uterine perforation. Three patients 5.7% developed septicaemia due to uterine gangrene. Ten patients 19.2% had acute pelvic infection presented with pelvic abscess and acute pelvic inflammatory disease. Maternal motility in our study was 2 out of 52 (3.8%). CONCLUSION: Induced septic abortions contribute significantly to maternal morbidity and mortality. Improving literacy rate in our female population and effective family planning should reduce its incidence. Different resources should be used to develop awareness of the hazards of induced abortions in the community. PMID- 19999230 TI - Is nitrous oxide necessary for general anaesthesia? AB - BACKGROUND: Nitrous oxide (N2O) has been used for about 150 years in clinical anaesthesia. Several recent reviews of the effect of nitrous oxide have concluded that there are certain contraindications to the use of this gas for general anaesthesia and its ecological effects, ozone depleting potential, immune depression and the proven factor of PONV have questioned the routine use of nitrous oxide in patients undergoing surgical procedures in general anaesthesia. METHODS: This study comprised of 200 adult patients undergoing general anaesthesia with 40% O2 and Sevoflurane with and without N2O. All patients had standard anaesthetic care and monitoring with BIS monitoring in 120 patients. The effect of avoiding N2O was observed on anaesthetic perioperative management and haemodynamics, PONV and pain in PACU. RESULTS: Demographic and perioperative characteristics were similar to both groups. Nitrous oxide free group needed only 0.233% (mean) more Sevoflurane. There was a marked reduction in incidence of PONV (11% to 5%) in N2O free group. Duration of surgery (97.72 +/- 52.393 in N2O group, 103.75 +/- 48.671 in N2O free group) and induction dose of propofol (155.30 +/-38.572 in N2O group and 158.50 +/- 36.164 in N2O free group) did not differ significantly in the two groups. CONCLUSION: The omitting of N2O from anaesthetic regimen has a substantial impact on patient comfort after surgery by reducing incidence of PONV and it does not have any justifiable indication of its use in General anaesthesia. PMID- 19999231 TI - Internet access and utilization for health information among university students in Islamabad. AB - BACKGROUND: Internet has changed the way we live and work. Advent of this technology has fundamentally transformed our lives the way invention of automobile changed how our lives and cities looked and worked before. Practically no information is available on the use of Internet for health by the people of Pakistan. The Objectives of the study were to assess the access and utilization pattern of Internet by university students in Islamabad, with emphasis on the healthcare information seeking. METHODS: An anonymous, self-administered, and pre tested questionnaire with questions on the access, and usage pattern of Internet, seeking health care information online, and belief about reliability of such information; was distributed to only those students who were enrolled in masters or higher degree programs. RESULTS: A total of 600 students were approached and 598 (99.7%) completed the questionnaires. The mean age of students was 23.5 years (range 19-40). The majority of students (423) were enrolled in masters program. Four hundred and sixty-eight students (78.26%) students had access to the computer either at home or at their university hostel. While 304 (50.84%) students had Internet access at home or in their university hostel. Out of 304 students who reported having access to Internet in the past three months, one hundred and thirty-nine (43.4%) students replied affirmatively to the question of having used Internet for seeking health care information. And 109 (78.4%) thought that such information was reliable. Out of 139 students who had used Internet for seeking health information, 35 (25.2%) students replied affirmatively to the question of having discussed health information obtained from Internet with their doctor/physician whom they visited for any illness/treatment. CONCLUSION: Majority of Islamabad university students in this study had access to computer and Internet. Young and healthy state of this educated age group perhaps accounts for limited use of Internet for seeking healthcare related information. However, high reliability of Internet obtained health information needs to be further studied in terms of websites utilized for seeking such information. PMID- 19999232 TI - The cradle of the deltaF508 mutation. AB - Cystic fibrosis (CF) is the most common autosomal recessive disorder caused due to mutation/s in the CFTR gene. The most common mutation in CFTR worldwide is deltaF508 and cystic fibrosis genetic analysis consortium revealed that this mutation is responsible for approximately 66% of all CF chromosomes in the world. Studies looking at the DNA polymorphic haplotypes created by CF linked markers suggest that deltaF508 has a single origin as this mutation has been found associated exclusively with one marker haplotype. Despite a high prevalence of this mutation in CF patients in northern parts of Europe, findings suggest that this mutation was not spread by Europeans but by a group that is speculated to have originated in the Middle East or a more eastern region in Asia (most likely subcontinent). Over here we have given a brief introduction to cystic fibrosis and classification of CFTR mutations and have further elaborated on the crucial issue about the spread of the deltaF508 mutation. We have reviewed findings that give clues about the origin of this mutation from the Baluch ethnicity residing in Pakistan. PMID- 19999233 TI - Retroperitoneal masses: different clinical scenarios. AB - We present two cases of retroperitoneal masses with different presentations and outcomes. The first case was a 22 years old primigravida lady who underwent emergency caesarean section for preterm premature rupture of membranes with breach. On the operating table, a large retroperitoneal mass was identified and the biopsy confirmed Burkitt's lymphoma. Post operative chemotherapy did not have a favourable result and the patient had a fatal outcome. The other case was a 15 years old boy who had a progressively increasing retroperitoneal mass. Exploratory laparotomy revealed a hard, fixed, unresectable tumour extending into the mesentery of the small gut, biopsies were taken which showed tuberculosis. Post operative antituberculosis treatment had a marked response and the tumour disappeared after 6 months. PMID- 19999234 TI - An unusual side effect of interferon alfa 2A: digital clubbing. AB - Interferon Alfa has been widely used to treat chronic hepatitis C virus infection. In this report, we present a case series of two patients referred to Sarwar Zuberi Liver Centre, Civil Hospital Karachi, who suffered from chronic hepatitis C. After getting detailed clinical examination and baseline work up prior to starting treatment, these patients were offered therapy with usual recommended dose of 3 million units of alpha-interferon subcutaneously thrice weekly. Both these patients developed clubbing of fingers during the course of treatment, one developing it during the 2nd month while the other during the 4th month. It was of grade II in one patient and of grade III in another and was bilateral in both the cases. Clubbing was not presented prior to start of treatment and no other secondary cause of clubbing was found in any of the case. These patients were not on any other drug that is known to interfere with interferon or can be associated with clubbing. No national or international data regarding such unusual side effect is available. Whether this effect is idiosyncratic or dose related and whether it is reversible or not after completion of treatment is yet to be established. PMID- 19999235 TI - Two-port laparoscopic cholecystectomy--a new technique. AB - Laparoscopic cholecystectomy is now the procedure of choice in all the gall bladder diseases and there is increase in the skills of surgeons with newer equipment. Normally the laparoscopic cholecystectomy is performed by using four ports or three ports. Two ports laparoscopic cholecystectomy is rarely performed as it demands greater expertise and skills. Also this technique is less expensive and less scar formation than four port laparoscopic cholecystectomy. We present a case of a middle aged female who underwent successfully laparoscopic cholecystectomy by using only two ports. PMID- 19999236 TI - Reducing and eliminating neuropathic pain. AB - Neuropathic pain is initiated or caused by a primary lesion or dysfunction in the central and/or peripheral nervous systems, including infection, trauma, metabolic abnormalities, and nerve compression, and is typically accompanied by hyperalgesia and allodynia. Neuropathic pain can be mild to excruciating, debilitating, difficult to manage, cause depression, decrease the quality of life, require extremity amputations, and has a variety of clinical symptoms. It effects up to 5% of the population, 70% of patients with advanced cancer and inflammatory pathologies, and 95% of patients with spinal cord injuries. The primary treatments of neuropathic pain are antidepressants, anticonvulsants, local anesthetic/topical agents, and opioids. The rapidly evolving symptom- and mechanism-based approaches to the treatment of neuropathic pain holds promise for improving the quality of life of patients with neuropathic pain. However, pharmacological treatment of the symptoms are difficult because of the limited understanding of the underlying causes of the pain, and the systemic levels of multiple side effects induced by various agents at an effective dose. Further, neuropathic pain is often refractory to conventional analgesic treatments, with most patients obtaining only partial relief with these agents, and with tolerability or side effects often limiting their use. Alternative treatments to pharmacology include peripheral or neuraxial nerve blockade, and implanted cortical or spinal cord stimulators. However, the great need remains for development of new and more effective approaches to reducing neuropathic pain. This review examines various approaches currently used for treatment of neuropathic pain and potential new and more effective approaches. PMID- 19999237 TI - Outcome of liver transplantation in a Hispanic population: 100 liver transplants in Puerto Ricans. AB - BACKGROUND: The residents of Puerto Rico (PR) had limited access to liver transplantation (LTx) prior to 1996. LTx remains locally unavailable and success rates for LTx for patients from PR have never been published. The outcome of the first 100 LTx recipients from PR transplanted at our center is analyzed. METHODS: 100 consecutive patients transplanted between 3/1997 and 1/2005 were evaluated. RESULTS: Hepatitis C was the indication for LTx in 44%. Overall patient survival at 1, 3 and 5 yrs was: 94.0%, 81.4% and 75.7%, respectively, while for hepatitis C, it was 90%, 73% and 73%, respectively. At mean follow up of 44 mo., 80% of patients were alive (66% HCV were alive vs 91% non HCV, p < 0.01). CONCLUSIONS: Access to LTx in Puerto Rico has dramatically improved since 1996. The government sponsored fund has provided access to indigent patients. Decreased survival in this minority population was not observed at 1, 3 and 5 years. Long-term survival was most affected by recurrence of HCV. PMID- 19999238 TI - Predictors of school dropout among adolescents in Puerto Rico. AB - OBJECTIVE: This research aims to understand the circumstances associated with school dropout in a cohort of Puerto Rican adolescents. METHODS: The study collected data from adolescents and their parents. Information related to school dropout among adolescents was obtained from the second year follow-up data from the longitudinal study funded by NIDA "Risky Families Embedded in Risky Environments" (Grant No. R01 DA 15301). Data was collected employing a self administered and a face-to-face interview protocol. Prediction of school dropout was assessed throughout adolescent characteristics, family background, school experiences and behaviors. RESULTS: During the second follow-up, two years after the baseline assessment, approximately 6.2% of the adolescents reported dropping out from school. Logistic regression analysis indicates that older adolescents (OR = 6.6, 1.37-31.67), whose mother used drugs during pregnancy (OR = 4.9, 1.31 17.91), who reported high rates of absenteeism (OR = 4.8, 1.63-14.13), high school grade retention (OR = 3.7, 1.14-12.05), and attended school where teachers were attacked or wounded by students (O R =7.0, 1.44-34.17) were more likely to dropout of school. DISCUSSION: : These findings emphasize the need to further understand the effects of different elements of adolescents' environment such as family and school. It has been posited that dropping out of school is a process whose characteristics can be detected long before it occurs. The fact that students who dropout are more likely to report skip classes and grade retention can be relevant elements in prevention and early intervention for teachers and other school personnel. PMID- 19999239 TI - Endoscopic third ventriculostomy: a surgeon's experience. AB - INTRODUCTION: Endoscopic third ventriculostomy (ETV) is considered an alternative treatment for certain types ofhydrocephalus. Depending on patient's age and etiology of hydrocephalus, it carries a success rate of around 90%. However, as in any surgical procedure, inherent risks are present and a risk-benefit analysis must be done prior to selecting patients for this intervention. OBJECTIVE: To evaluate retrospectively the experience of ETV at the University of Puerto Rico and examine the etiological factors, demographic data and symptoms among the Puerto Rico population. This study represents the data of one neurosurgeon and is the first account of endoscopic third ventriculostomy as a mode of treatment in the Caribbean area. METHODS: Retrospective analysis was undertaken of 29 patients treated at the University Pediatric Hospital (UPH) and University District Hospital (UDH) in San Juan was undertaken Different etiologies of hydrocephalus were identified and managed. RESULTS: The most common indication for ETV was aqueductal stenosis (59%) with male predominance (55%). Also, the most common population treated were adults (72%) and the most common symptom presented were headaches (52%) and gait disturbances (43%). CONCLUSIONS: Endoscopic third ventriculostomy is a safe and effective way to treat hydrocephalus and should be considered as first choice of treatment for certain patients with hydrocephalus unless otherwise contraindicated. PMID- 19999240 TI - Descriptive profile of surgically-confirmed adult central nervous system tumors in Puerto Rico. AB - INTRODUCTION: Published studies regarding the incidence of central nervous system (CNS) tumors in Puerto Rico (PR) are exceedingly rare. The general understanding is that the incidence of these tumors in Puerto Rico is similar to the one found in the United States of America (USA). The objective of this study is to describe the specific profile of all the CNS tumors that are surgically intervened in Puerto Rico, through the creation of a database. METHODS: A retrospective analysis of all the surgical procedures from January 1, 2002 to May 31, 2006 for adult CNS tumors in Puerto Rico was performed. Each case was evaluated for demographic information, operative procedure, lesion description and official pathological report. Recurrent lesions were excluded. The information was organized to form a database of all the CNS neoplasms. RESULTS: A total of 1,018 procedures for CNS tumors were performed on 1,005 patients. The incidence rate of surgically intervened CNS tumors in Puerto Rico is 6 per 100,000 people. CNS tumors were more common in women than in men (58% vs. 42%), respectively. The mean age was 52.4 years. The most common histological type found was meningioma WHO I (24%), followed by pituitary adenomas (16%), and glioblastoma multiforme (14%). CONCLUSIONS: Our results reflect a unique histopathological distribution of operated CNS tumors in Puerto Rico. In this series, primary tumors are more common than metastatic tumors. Benign histological tumors were more frequent than more malignant variants. Although this study reflects only the histologically diagnosed tumors, it is headway towards diagnosing the incidence of all CNS tumors in Puerto Rico. PMID- 19999241 TI - TRAMIL ethnopharmacological survey: knowledge distribution of medicinal plant use in the southeast region of Puerto Rico. AB - BACKGROUND: TRAMIL network aims to understand, validate and expand health practices based on the use of medicinal plants in the Caribbean, a "biodiversity hotspot" due to high species endemism, intense development pressure and habitat loss. OBJECTIVES: The purpose of this study was to document both the medicinal plants that are frequently used to treat health conditions prevalent in the southeastern region of the archipelago of Puerto Rico and the trends in their use among the study population. METHODS: An ethnopharmacological survey was conducted in the study region. The results were analyzed using univariate and multivariate statistical analyses. RESULTS: Overall, 118 medicinal plants were recorded as being used to treat depression, nervousness, chronic sinusitis, gastritis, gastroesophageal reflux disease, allergic rhinitis, rhinofaryngitis, asthma, arthritis and migraine. The plant species with significant use were Citrus aurantium L., Citrus aurantifolia (Christm.) Swingle, Pluchea carolinensis (Jacq.) G. Don in Sweet, and Mentha piperita L. The use of medicinal plants is more frequent among single women with a high educational level, a trend similar to the use of CAM in the US. CONCLUSION: Ethnopharmacological knowledge and the use of medicinal plants is decreasing in the study region due to an increase in the use of conventional medical care and to self-medication with over-the-counter pharmaceutical products. Four botanical species with significant uses that were not previously recorded in the Caribbean Herbal Pharmacopoeia have been identified. This report will be followed by the scientific validation and toxicity studies of these plant species and the TRADIF activities in the study region. PMID- 19999242 TI - Exploratory study: barriers for initiation and/or discontinuation of breastfeeding in mothers of children with Down syndrome. AB - BACKGROUND: The aim of the study is to identify the barriers associated with breastfeeding in mothers of Puerto Rican children with Down. METHOD: A non probabilistic sample (n = 26) of mothers was used in the study. The sample was obtained in an institution in Puerto Rico specializing in care of infants with Down Syndrome [IDS]. A self-administered questionnaire was used consisting of six sections. Descriptive statistics were used for data analysis. RESULTS: The majority of interviewed mothers (80.8%) had 70.0% or higher correct answers regarding the benefits of breastfeeding. 84.6% of the mothers who decided not to breastfeed or who discontinued breastfeeding reported as the main reason that the baby presented sucking problems and 50.0% of the participants were not allowed to have their baby with them during their stay in the hospital. 84.3% of the participants indicated having breastfed their IDS and 61.5% stated that they would not breastfeed in public places. CONCLUSION: Active support, instruction, and collaboration among members of the health care team are essential for breastfeeding success. PMID- 19999243 TI - Medicine in the 21st century: towards a Darwinian medical epistemology. AB - In this review we reflect upon the new science of Darwinian medicine. It is a tenet of modern biology that evolutionary theory as proposed by Charles Darwin and further refined via the new synthesis is the common thread that ties all of biological inquiry into a coherent whole. This review aims at making clear how evolution by natural selection is relevant to medicine. A set of pertinent examples linking Homo sapiens sapiens' present disease conditions to its evolution during the Late Pleistocene and Holocene epochs are discussed. The review concludes with observations as to the epistemological value of evolutionary theory as a heuristic tool for articulating a medical paradigm in accord with modern biology. PMID- 19999244 TI - Medical milestone, millennial mission. PMID- 19999245 TI - Awake laminotomy and discectomy using Dexmedetomidine as anesthetic. AB - Lumbar discectomy is the most common surgical procedure performed in the United States for patients having back and leg symptoms. The vast majority of the procedures are elective and are performed under general anesthesia. This is a reported case of a 40 year old woman with worsening symptoms of L5-S1 disc herniation unresponsive to conventional therapies. The use of Dexmedetomidine was discussed with the patient, explaining the advantage of motor and sensory testing during the surgery and immediate patient feedback. Awake laminotomy and discectomy could be performed as an ambulatory procedure, since the use of Dexmedetomidine as the main anesthetic represents a safe and convenient alternative to general anesthesia in this patient with lumbar disc herniation. PMID- 19999246 TI - Annular erythema associated with anti-Ro/SS-A and anti-La/SS-B antibodies: a new case in a Hispanic patient. AB - We describe an 18 year-old male patient who was anti-Ro/SS-A and anti-La/SS-B positive and presented with recurrent annular plaques on the trunk, arms, face, and scalp with evidence of associated patchy alopecia. The skin biopsy revealed a superficial perivascular and periappendegeal lymphocytic infiltrate and focal areas of vacuolar alteration and smudging of the dermoepidermal junction. The patient also presented with a history of xerophthalmia. Skin lesions as well as sicca symptoms responded to antimalarial treatment with hydroxychloroquine. This case demonstrates a new case of annular erythema indistinguishable from those previously described in patients of Asian descent occurring in a Hispanic patient. PMID- 19999247 TI - Global payment system no Massachusetts miracle. PMID- 19999248 TI - Everybody wants to close the doughnut hole. PMID- 19999249 TI - Marginal increase in chief medical officer pay. PMID- 19999250 TI - How patient-centered medical homes may change U.S. medicine. PMID- 19999251 TI - Will the new fat-fighter drugs be more worthy of coverage? PMID- 19999253 TI - Research topics underpin comparative effectiveness. PMID- 19999252 TI - A conversation with James Sabin, MD. When values clash. PMID- 19999255 TI - Retail pharmacies miss out on specialty drugs. PMID- 19999254 TI - Effect of social work intervention on hospital discharge transition planning in a special needs population. PMID- 19999256 TI - Review panels pay off for Colorado-based plan. PMID- 19999257 TI - Replace all those sample closets with high-tech dispensing machines. PMID- 19999258 TI - Shortage of chest surgeons likely by 2020. PMID- 19999259 TI - Building a statewide clinical trials network for cancer care in West Virginia. AB - In the United States, mortality rates have been declining for certain tumors, For the majority of advanced stage cancer types, cure is unattainable but treatment is still evolving. Advances in the treatment of cancer can be achieved by enrolling patients in cancer clinical trials. Presently, less than 3% of adult cancer patients participate on clinical trials in the United States. Providing cancer care and access to clinical trials are a challenge in a rural state, with a dispersed population base, such as West Virginia. Building upon recognition of barriers to clinical trials awareness and access, oncology leaders in the state are in the formative stages of developing a statewide cancer clinical trials network. Realization of this network will have an enormous impact on cancer care in our state and perhaps can serve as a model for other community and physician teams for other diseases. PMID- 19999260 TI - Psychosocial oncology services in West Virginia. AB - While the field of psychosocial oncology has been recognized for several decades, specialized psychosocial services to West Virginians with cancer have been limited. Considering the increase in the incidence of cancer and the recognized burden of cancer-related emotional distress, healthcare providers must screen for emotional distress as the sixth vital sign, and develop an array of evidence based interventions and services for patients with cancer and their families along the cancer trajectory. PMID- 19999261 TI - Current concepts in breast reconstruction. AB - Breast cancer affects many women, but with advances in detection and treatment, survival rates have increased. Thus, it is important to understand that there are many reconstructive options available to help ease the psychological burden of mastectomy. Reconstructive options include tissue expander/implants, biologics, and several autologous tissue options, including pedicled latissimus and TRAM flaps, free TRAM flaps, and perforator flaps. We present a discussion of reconstructive techniques, the risks and benefits of each, and individual patient considerations that will help physicians to guide treatment options. PMID- 19999262 TI - Breast cancer screening, incidence, and mortality in West Virginia. AB - Situated entirely in the Appalachian region, West Virginia (WV) as a state is characterized by a diverse population, with a major portion of the population residing in rural areas. Breast cancer is the most commonly diagnosed cancer among women residents of the state. Using different data sources, we have reported the most recent screening, incidence, and mortality information associated with breast cancer in WV. In 2006, roughly one-fourth of women > or =40 years of age had not had a mammogram in the past two years. Sociodemographic factors were found to be associated with lack of mammography screening among these women. The leveling off of the increase in the in-situ breast cancer incidence rates in WV since 1999 is closely associated with the bottoming out of the proportion of women without mammography that has been seen since early this decade. Counties in southern and northern part of the state generally have higher invasive breast cancer incidence rates than those in the central part. Breast cancer-related mortality is higher among southern counties in comparison to counties in northern and central part of the state, with mortality rates higher than the national average in McDowell, Mingo, Wayne, and Mason Counties. There is a compelling need to increase access to mammography screening among women residents in the state. PMID- 19999263 TI - Use of radiation after breast conserving surgery (BCS) for DCIS and early invasive breast cancer at Charleston Area Medical Center (CAMC). A study of compliance with National Comprehensive Cancer Network (NCCN) guidelines. AB - The National Comprehensive Cancer Network (NCCN) recommends that patients with ductal carcinoma in situ (DCIS) and stage I/II invasive breast cancer receive radiation therapy following breast conserving surgery (BCS). The purpose for our study was to determine 1) the percentage of patients with DCIS and stage I/II breast cancer who received radiation therapy following BCS and 2) the clinical factors associated with the use of radiation. We retrospectively studied 606 patients treated between 2000 and 2007 with BCS for DCIS (n=104) and stage I/II breast cancer (n=502). Overall 93 percent of patients in our study received radiation therapy. We found that almost 85 percent and 95 percent of patients with DCIS and stage I/II breast cancer respectively received radiation therapy. Patients with invasive breast cancer who were less than 70 years of age and who received adjuvant systemic therapy were significantly more likely to receive radiation. The data from our study indicate that the use of radiation following BCS is high at our institution. Periodic review of treatment practices at local hospitals is valuable in assessing compliance with national guidelines and in improving quality of care. PMID- 19999264 TI - A physician perspective: belonging to that "special club". PMID- 19999265 TI - Breast cancer in South-Central West Virginia. PMID- 19999266 TI - Smoking and breast cancer screening in West Virginia: opportunities for intervention. AB - Smoking has been increasingly identified as a risk factor for breast cancer among women. West Virginia has high rates of smoking, which can be further examined in relation to breast cancer screening and smoking related variables. This study used 2006 Behavioral Risk Factor Surveillance System (BRFSS) data to examine the relationship between smoking related variables and breast cancer screening behaviors among women in West Virginia age 40 and older. Findings showed that approximately 21.6% of women in the sample were current smokers, with a majority of these women smoking every day (82.5%). Half of the women in the study sample had made at least one past year quit attempt. Current smokers were less likely to have had breast cancer screening in the past one or two years, and among these women who smoke, every day smokers were less likely to obtain breast cancer screening. Smokers who did not make a quit attempt during the past year were also significantly less likely to have had mammography screening. Study results highlight an unmet need and opportunity for intervention related to breast cancer screening among women who smoke, and especially those with higher smoking intensity. Smoking status and smoking intensity should be recognized as predictors of women who are less likely to obtain breast cancer screening. These women can be identified and targeted as an important high risk population with unmet need for smoking cessation and breast cancer screening interventions. PMID- 19999267 TI - Triple-negative breast cancer in West Virginia. AB - In 2007, the American Cancer Society ranked West Virginia 43rd in breast cancer incidence rates for individual states. Despite our improvements in medical care, the advanced pathological characteristics of breast cancer at diagnosis receive little attention. Consequently, we compared the changing pattern of early breast cancer in several cohort studies conducted at regional medical centers in West Virginia. The data used in this analysis was derived from 320 women presenting at West Virginia University Hospital (WVUH) in Morgantown between 1999 and 2004, with a diagnosis of invasive breast cancer. Details of age, tumor size and axillary lymph node status were compared with tumor registry information published from a cohort study of 191 patients from the Charleston Area Medical Center (CAMC) between 1990 and 1991. Only histologically documented adenocarcinomas of the breast were included. Tumor size was characterized using the TNM system and staged according to AJCC criteria. For comparative purposes, details from the two regional centers were compared with tumor characteristics from a large longitudinal cohort of 2,484 breast cancers from the Women's Health Initiative (WHI) study. Baseline median age at diagnosis of women screened at WVUH was younger than patients at CAMC (52 vs. 60). Women diagnosed with triple negative breast cancer at WVUH and CAMC had similar age distributions. Within the triple-negative patients at WVUH, 44% of patients were less than 50 years of age and 20% were less than 40 years of age. At CAMC, 35% were less than 50 years of age and 7% were less than 40 years of age. For women at WVUH, 61.5% presented with T1 tumors compared to 65.5% at CAMC. These figures were lower than the WHI average of 80.3%. In contrast, more women presented with larger T2 tumors at our medical centers compared with the national study, 32.6% versus 17.4% respectively. At WVUH, 2.3% of women had T3 tumors (> or =5 cm) compared with 1% at CAMC. Similar to the WHI study, 35-42% of women at WVUH and CAMC were diagnosed at the T1c stage. Approximately, 30% were diagnosed with positive lymph nodes, compared to 23% in the national study. Combined breast cancer data from our medical centers show an increase in more advanced tumors and positive regional lymph node involvement at the time of diagnosis compared to national reports. Other factors such as obesity, diabetes, poverty and access to mammography screening could be influencing the poorer outcomes for women with breast cancer in West Virginia. PMID- 19999268 TI - Perspectives on cancer health disparities in West Virginia. AB - "Perspectives on Cancer Health Disparities in West Virginia" provides an overview of the factors that lead to health disparities in general, to the burden of breast and other cancers in our state, and highlights the deep-rooted values and characteristics that will help communities and their partners achieve parity. PMID- 19999269 TI - The role of ultrasound in breast imaging. AB - This report describes several patients in whom diagnostic ultrasound played a significant role in the detection and diagnosis of breast masses. Diagnostic ultrasound was able to verify whether breast mass was or was not present. Secondly, ultrasound was able to determine whether the mass was definitely benign and no further evaluation was necessary, or whether the mass was possibly malignant and needed a biopsy. The important role diagnostic ultrasound plays in breast imaging and its appropriate clinical applications are discussed. PMID- 19999270 TI - Bonnie's Bus--cancer disparities in West Virginia, philanthropy and opportunities to build lasting partnerships. AB - The State of West Virginia (WV) has significant cancer health disparities, substantiated by the fourth highest cancer mortality rate in the nation. In October 2007, Jo and Ben Statler donated $5 million dollars to the Mary Babb Randolph Cancer Center to establish a mobile mammography program (Bonnie's Bus) and endowed research positions at West Virginia University. This opportunity paved the way for an additional $2.5 million dollars of funding through a match program afforded the by WV Eminent Scholars Program and $700,000 of private foundation (Susan G. Komen for the Cure and Claude Worthington Benedum) support to begin to craft a statewide clinical trials network linked to Bonnie's Bus to provide new access to state-of-the-art clinical trials for residents of WV. Philanthropic gifts, as illustrated by the Statler gift, provide momentum for academic research and health care programs and new opportunities await through another legislative initiative--the WV Research Trust Fund. The Mary Babb Randolph Cancer Center is working to address cancer health disparities in the state of WV. PMID- 19999271 TI - From high tech to high touch: integrating community voices in mobile mammography outreach. AB - Providing mobile screening mammography services across the state of West Virginia (WV) presents unique challenges. The ability of new machines to screen for breast cancer is modern medicine at its best. The use of the mobile unit, "Bonnie's Bus", holds promise for getting this high tech equipment to women in rural areas of West Virginia where it is most needed. Cancer detected in early stages is more treatable and women have a better chance of becoming survivors. The key to the program's success resides with the women who come for the screening, the extension network of community members who help set up and assist women to access the screening system, and community health care providers who care for the women if cancer is found. Linking "community voices" with the programs funded under the WV Komen grant screening programs provides a winning solution for West Virginia and the women served by Bonnie's Bus. Information obtained from five focus groups of 58 community leaders and women residing in rural WV was used to develop the mobile program. Building upon established relationships with the West Virginia Breast and Cervical Cancer Screening Program (WV BCCSP) coordinators and providers, outreach and information gleaned from WV communities, is being used to develop community partnerships based on mutual trust and respect to advance the common goal of decreasing breast cancer related illness and death in West Virginia women. PMID- 19999272 TI - Stroke outcomes. PMID- 19999273 TI - [Operation treatment and medicine treatment of type 2 diabetes mellitus combined with obesity]. PMID- 19999274 TI - Semi-quantitative cultures of endotracheal aspirates still commonly used for VAP. PMID- 19999276 TI - [Obsessive-Compulsive disorder: present and future aspects]. PMID- 19999275 TI - "Do grandmas have husbands?" Generational memory and twentieth-century women's lives. AB - This essay uses memory in the ancient and modern sense of the "inner life of thought" to describe the formation of generational memory in a modern professional family whose twentieth-century history has been fractured by migration, war, education, and divorce. It is about the power of feeling and law, which framed the practical freedoms of twentieth-century women's lives and introduced the modern citizen in the aftermath of universal suffrage and world war. The first part of the essay emphasizes the psychic dimension of bodily feeling and drive in the formation of memory; a dimension overlooked by oral history and social movements, yet confirmed by autobiography and memoir. My granddaughter's questions provoked resistance as well as family stories, and let me observe the thought process in a child. Social history, autobiography, and personal memory confirm the common experience of everyday life reaching back through generations of London families; folklore, commerce, and family story make narratives of dreams, hopes, terrors, and events; a child's comprehension of the outside world is grasped through curiosity, imagination, and play in which bodily feeling is as powerful as speech and prohibition to make meanings that flow between inner world and external reality. The second half of the essay reflects on Joan Riviere's description of the self. Leading British psychoanalyst, translator of Freud, writing in the 1950s, Riviere's language of the inner world resonates with the liberal social ethics -- empathy, public service, common good - which underpinned women's and human rights mid-twentieth century and the egalitarian and reproduction reforms whose universalism has been challenged since the 1970s. Negative feeling is striking in Riviere's description of the self -- fear, shame, shock, and trauma, which are confirmed in memoir and autobiography. In contrast, liberal social democratic accounts of the time idealized English character. Today, the future uncertain, memory -- in the ancient and modern sense of the "inner eye of thought" -- is more than ever necessary for social movements as for the individual. PMID- 19999277 TI - Diasporic memories: community, individuality, and creativity -- a life stories perspective. AB - Can we talk of a collective, diasporic memory? I will argue that in the case of the African-Caribbean community, there are distinctive features -- such as the need to "tell" and the need to "connect" -- which suggests that this diasporic memory is framed through identifiable cultural templates, which distinguish it from the memories of migrants. PMID- 19999278 TI - [Change in diagnosis of and reconsideration for the concept of obsessive compulsive disorder]. PMID- 19999279 TI - [Physiopathology of obsessive-compulsive disorder--special reference to aspects of the neurochemistry]. PMID- 19999280 TI - [Update on physiopathological study of obsessive-compulsive disorder; the neuroimaging study]. PMID- 19999281 TI - [Tic-related obsessive-compulsive disorder]. PMID- 19999282 TI - [Trends in diagnosis and classification of obsessive-compulsive disorder: DSM-V]. PMID- 19999283 TI - The web of clinical trial registration obligations: have foreign clinical trials been caught? AB - The web of overlapping requirements, standards, recommendations and policies governing the conduct of clinical trials highlights the intense scrutiny of the ethical, data quality and public access issues raised by human trials that are conducted to demonstrate the safety and efficacy of medical products marketed in the United States. One relatively recent development is the requirement that sponsors register and make public information about their clinical trials and clinical trial results. These clinical trial registration requirements illustrate the interests of patients, providers and researchers in increased visibility, transparency and accessibility of clinical trials and the data they generate. These requirements, however, pose regulatory, logistical and practical hurdles for companies sponsoring clinical trials of drugs and medical devices. PMID- 19999284 TI - Regulation of smoking cessation drugs by the Food and Drug Administration. AB - There are about 50 million active cigarette smokers in the United States, and cigarette smoking accounts for almost 500,000 deaths each year. The World Health Organization (WHO) places the global total of such deaths at 5 million people annually. In the United States, this high-level of cigarette smoking occurs despite the fact that 70 percent of Americans who smoke report that they want to quit the habit. As a direct consequence of these statistics, several smoking cessation aids, some as the result of the discovery and development of new molecular entities, have been developed both domestically and abroad. While the timely development and commercialization of new molecular entities serves a vital public interest, so too do assurances of their safety and efficacy. This article proposes an integrated approach early in the clinical development of a new molecular entity to the design of premarket approval clinical studies, postmarket approval clinical studies, and Risk Evaluation and Mitigation Strategies (REMS), as a means of balancing these two important policy objectives. PMID- 19999285 TI - [Mental disorders associated with alcoholism]. PMID- 19999286 TI - Life after Riegel: a fresh look at medical device preemption one year after Riegel v. Medtronic, Inc. AB - The United States Supreme Court's landmark decision in Riegel v. Medtronic, Inc. in February 2008 altered the landscape of medical device litigation involving Class III medical devices that have received premarket approval from the Food and Drug Administration (FDA).In Riegel, the Court ruled that the premarket approval process establishes federal requirements and that state law tort claims seeking to impose additional or different requirements on device manufacturers are preempted under the express preemption clause of the Medical Device Amendments (MDA) to the Food, Drug and Cosmetic Act (FDCA). It is only by examining post Riegel case law that one can develop an appreciation for where medical device litigation and device preemption are likely to head in the future. This article will conduct this evaluation of post-Riegel decisions by dividing the courts' rulings into various types of cases. This classification of claims provides insights into the strategies employed by plaintiffs to distinguish Riegel or side step the ruling altogether and how courts have received these efforts. PMID- 19999287 TI - Casting a global safety net--a framework for food safety in the age of globalization. AB - In mid-March 2007, Ontario-based Menu Foods Inc. started recalling its "cuts and gravy" style pet food, after receiving information that pets that had eaten the product had fallen ill. Within a week, the company was inundated with complaints and expressions of concern from about 200,000 of its customers. The Food and Drug Administration (FDA) determined in late March 2007 that the most likely culprit in the illness, and in some cases death of the pet animals, was contaminated wheat gluten, a vegetable protein imported from China. One of the FDA identified contaminants was an industrial chemical called melamine. Reports of widespread adulteration of animal feed with melamine in China raised concern of similar contamination in the human food supply. In response, on April 27, 2007, FDA announced the detention of all vegetable proteins imported from China, whether for animal or for human consumption. But, FDA's action came too late. On May 1, 2007, officials from FDA and the U.S. Department of Agriculture (USDA) indicated that between 2.5 to 3 million people in the United States had consumed chickens that had been fed with contaminated vegetable proteins imported from China. The 2007 pet food recall incident provided an ominous early warning that, unless the international community can come up with a better food safety mechanism, more such food contamination disasters could happen in the future. PMID- 19999288 TI - A new history and discussion of 180-day exclusivity. AB - Congress created 180-day exclusivity for generic drug applicants in the 1984 Hatch-Waxman amendments to the Federal Food, Drug, and Cosmetic Act (FDCA) and amended the provision substantially in the Medicare Prescription Drug, Improvement, and Modernization Act of 2003 (MMA). The fundamental goal behind 180 day exclusivity was to provide an incentive for generic drug applicants to challenge innovator patents, and the core of the concept--as it has been applied by the Food and Drug Administration (FDA) and the courts--is that the first generic drug applicant to challenge an innovator's patent is entitled to six months of exclusivity against subsequent patent challengers for the same innovator drug. 180-day exclusivity is governed by sections 505(j)(5)(B)(iv) and 505(j)(5)(D) of the FDCA. In this article, the authors provide a comprehensive resource on 180-day exclusivity for old abbreviated new drug applications (ANDAs) (but less detail in some places where the 2007 article may be referenced) but focus more discussion on the new provisions as well as some policy and legal issues related to 180-day exclusivity that have not previously addressed. PMID- 19999289 TI - Off-label use reimbursement. AB - In this study, the authors examine factors underlying payer off-label use reimbursement policies. Updating a study published 14 years ago, presenting the results of the survey of 179 payers administering public (Medicare and Medicaid) pharmacy benefits. The focus is on payers administering pharmacy benefits in the public sector for two reasons: First, transparency; there is a tendency for such payers to reveal more about their decisionmaking processes than payers that exclusively deal with the commercial market. Second, Medicare and Medicaid set the pace in terms of specific policies on off-label use reimbursement, particularly with regard to anti-cancer agents and biologics. The authors investigate the role of primary and secondary sources governing reimbursement of unlabeled indications, including biopharmaceutical compendia, peer-reviewed literature, and clinical practice guidelines. The findings point to the continued variation of off-label use reimbursement policies across payers. Furthermore, the survey data shed light on payer efforts to design off-label use reimbursement strategies, including the use of cost-effectiveness. PMID- 19999290 TI - [Alcoholism and depression: catecholamines and brain-derived neurotrophic factors]. PMID- 19999291 TI - Homeland security planning: what victory gardens and Fidel Castro can teach us in preparing for food crises in the United States. AB - Two historical examples provide important insight into how federal government policies can integrate regional and local food systems to achieve food security during a time of acute crisis. During World War II, American home gardeners, through the federal government's Victory Garden program, supplied 40 percent of the nation's fresh produce, while simultaneously maintaining pre-war commodity production policies favoring large agricultural interests. The recent food crisis in Cuba, precipitated by the collapse of Soviet-bloc trade in the early 1990s, is another historical example that could inform U.S. policymakers on how to achieve food self-sufficiency through reemphasis on small farmers using sustainable practices supplemented with urban gardening. This article aims to ignite government action to strengthen and integrate regional and local food systems into federal food security planning so that citizens can be best prepared for a food emergency. The article first examines laws, regulations and policies put in place during World War II that employed regional and local food networks to satisfy a significant amount of civilian food supply needs. The article also looks at more recent Cuban efforts to achieve forced food self-reliance when, after the end of the Cold War, Soviet subsidies and preferential trading of energy and food supplies ceased almost overnight. PMID- 19999292 TI - [Alcohol drinking, alcoholism, and suicide]. PMID- 19999293 TI - [Alcoholism associated with anxiety disorders]. PMID- 19999294 TI - [Characteristics of and therapy for alcoholism with eating disorder]. PMID- 19999295 TI - [Alcoholism and cognition disorders]. PMID- 19999296 TI - [Psychiatric care for patients with addiction of amphetamine, CNS stimulant]. PMID- 19999297 TI - [Understanding of and therapy for eating disorders--psychiatric views]. PMID- 19999298 TI - [Schizophrenia and glutamate transport systems]. PMID- 19999299 TI - Reform of health: revolutionary or evolutionary? PMID- 19999300 TI - [Psychologic interview for the diagnosis of neuroses]. PMID- 19999302 TI - Treatment of uncomplicated malaria in Papua New Guinea: what should be done? PMID- 19999301 TI - [Therapy for alcohol abuse]. PMID- 19999303 TI - Knowledge and attitudes about infant feeding among nulliparous and parous women in Port Moresby: a comparative study. AB - Knowledge of the advantages of breastfeeding, the disadvantages of bottle feeding and the Papua New Guinean legislation to protect breastfeeding was determined in a comparative study of nulliparous and parous women attending the Antenatal Clinic of Port Moresby General Hospital. A high proportion (40%) of both groups had had experience of bottle feeding. Whilst the large majority (94%) of the women indicated that breastfeeding was the best way to feed babies, knowledge of the reasons for its superiority over bottle feeding and of the dangers of bottle feeding was poor in both groups. Knowledge of the legislation to protect breastfeeding was also poor and was the only area in which there was a significant difference between the groups, nulliparous women having poorer knowledge (p = 0.015). The level of education did not appear to be associated with knowledge about feeding. There is an urgent need to review the legislation, to find ways of enforcing it, and to improve the education of young people on issues of infant feeding. PMID- 19999304 TI - A review of the current state of malaria among pregnant women in Papua New Guinea. AB - Besides young children, pregnant women are at high risk of malaria in highly endemic countries. This paper reviews evidence from studies conducted in Papua New Guinea (PNG) in the last 20 years on the burden and prevention of malaria in pregnancy and highlights gaps in our knowledge of malaria in pregnancy in PNG. Overall, primigravidae were found to be at higher risk than multigravidae, with up to 40% of primigravidae but only 10-25% of multigravidae infected with Plasmodium falciparum at delivery. Such infections were found to be associated with a 128-145 g decrease in birthweight. Mean birthweights reported between 1980 and 2003 range from 2.58 to 2.72 kg in primigravidae and 2.84 to 3.09 kg in multigravidae, with 21% to 48% and 9% to 19% of babies born to primigravidae and multigravidae, respectively, of low birthweight (<2500 g). The negative impact of malaria in pregnancy is compounded by relatively low rates of antenatal coverage. The current PNG national treatment policy which prescribes a treatment course of first-line antimalarial treatment (currently chloroquine and sulphadoxine pyrimethamine) at first antenatal clinic contact, followed by weekly chloroquine prophylaxis and iron and folate supplementation, may no longer be effective given the high levels of resistance to chloroquine in PNG and poor compliance. In order to reduce the burden of malaria in pregnancy in PNG, alternative methods of control such as insecticide-treated nets and intermittent preventive treatment in pregnancy (IPTp), as well as improved modes of delivery of maternal health interventions, are urgently needed. PMID- 19999306 TI - Variations of the middle thyroid vein in Papua New Guinean Melanesians. AB - The middle thyroid vein has been noted to be frequently absent during thyroidectomies in Papua New Guinea (PNG). To verify this and other known inconsistencies in the neck a total of 103 bodies were dissected at the Port Moresby General Hospital in 2002. The middle thyroid vein was absent in 59% of the cases. It would be to the advantage of surgeons in PNG to be aware of this. PMID- 19999305 TI - A case-control study of VDRL-positive antenatal clinic attenders at the Port Moresby General Hospital Antenatal Clinic and Labour Ward to determine outcomes, sociodemographic features and associated risk factors. AB - Between June 2001 and December 2002, 152 antenatal patients at Port Moresby General Hospital who were Venereal Disease Research Laboratory (VDRL) serology positive and 150 unselected antenatal patients who tested negative were studied to determine the gestational age at which the tests were performed, the time it took for results to become available, the proportion of patients who received treatment, the sociodemographic characteristics associated with VDRL positivity and the effect of VDRL positivity on maternal and perinatal outcomes. The prevalence rate of VDRL positive among antenatal clinic attenders in Port Moresby at that time was 4.4%. Of the 152 VDRL-positive patients in this study 97% were also Treponema pallidum haemagglutination (TPHA) positive. Significantly more of the positive patients were of highlands origin, lived in settlements, had previous marriages, had lower parities, delivered preterm babies, had stillbirths, had growth-restricted babies and had babies with lower Apgar scores at both 1 and 5 minutes. The mean birthweight was significantly lower among the positive patients. Significantly more of the positive patients were married to spouses with occupations which were regarded as 'risky' for sexually transmitted infections. There was no difference between the two groups with respect to patient's education, marital status, husband's education, gestational age at delivery and the number of days the baby spent in the Special Care Unit. The study concluded that the current antenatal screening does not provide adequate coverage for our patients. If the current availability of clinic-based strip tests provided by a non-government organization can be continued by the Ministry of Health we should be able to overcome this problem. PMID- 19999307 TI - A focused ethnography about treatment-seeking behaviour and traditional medicine in the Nasioi area of Bougainville. AB - A focused ethnographic study examining knowledge, behaviour and attitudes related to treatment-seeking behaviour and traditional medicine was conducted in the Nasioi area of Bougainville. The study was conducted in two separate locations within the Nasioi area. Some similarities as well as some differences in knowledge and behaviour were exhibited in the two study locations which could be further investigated. People from both areas had a similar perception of common and serious illnesses. People from both areas also attributed illness to both biological factors and supernatural forces. Home management was the most common initial response to illness and the local village clinic or urban health centre were most frequently used as the first treatment resort. The most important factors in determining treatment-seeking responses to illness in both locations were cost and proximity to home or convenience. The study found that there may be considerable potential for an integrated health system in the Nasioi area and provided suggestions as to how this could be progressed. The study also provided information that could be useful in a wider context in informing the implementation of the national policy on traditional medicine in Papua New Guinea. PMID- 19999308 TI - Primary repair of colonic injuries at the Kundiawa and Madang General Hospitals, Papua New Guinea. AB - In this study, we evaluated the safety of primary repair of colon injury in a low volume tropical hospital setting. Between 1998 and 2005, 18 consecutive patients who underwent emergency operation for civilian traumatic colon injury were studied. The main outcome measures were the mortality and morbidity rates and the total length of the hospital stay. The mean hospital stay for one-stage repair was 12 days versus 29 days for the two-stage procedure, which was a significant difference (p = 0.009). There was no death reported from this study. There was no significant difference in postoperative septic complications between the one stage and two-stage procedures. One-stage repair of colonic injury is a safe and cost-effective option for selected patients in the tropical hospital setting. PMID- 19999309 TI - Barriers to the delivery of the hepatitis B birth dose: a study of five Papua New Guinean hospitals in 2007. AB - Hepatitis B is highly endemic in Papua New Guinea (PNG). Vaccination at birth is a key mother-to-child transmission prevention strategy. Despite recommendations for newborns to be vaccinated within 24 hours of delivery, a 2005 survey showed 23% coverage among children born in health facilities. Our study examined hepatitis B birth-dose coverage and knowledge, practices and barriers to vaccine delivery in five major PNG hospitals. Data on births and vaccines administered were sourced from the National Department of Health (NDoH) and directly from the five hospitals. A maternity unit audit and staff survey were undertaken. Across the five hospitals, the hospital-level data of hepatitis B birth-dose coverage was 79% (range: 40-96%) compared to 19% from national data (range: 0-106%). Despite hospitals having adequate vaccine supply, access to appropriately stored vaccine in maternity units was compromised with only one unit having a vaccine specific temperature-monitored refrigerator. In interviews of 25 staff, incorrect reasons given for delaying vaccination were prematurity (60%), low birthweight (48%) and difficult birth (36%). This study found encouraging birth-dose coverage rates in five major hospitals but 20% of babies still missed receiving the recommended vaccine. The NDoH Immunization Unit will use the results of this study to inform strategies to improve hepatitis B birth-dose coverage in hospitals. PMID- 19999310 TI - The prevalence of HIV infection in women attending antenatal clinics in Fiji. AB - HIV (human immunodeficiency virus) is an increasing concern in the South Pacific. We estimate, based on reported figures, that the prevalence of HIV infection in women attending antenatal clinics in Fiji in 2003 was 0.04%. The number of children born to HIV-positive mothers is small, though perinatal transmission appears to be high. Fiji's preliminary strategies for prevention of perinatal transmission have been significant, but require ongoing support and implementation. PMID- 19999311 TI - [Internal medicine, interdisciplinary medicine]. PMID- 19999312 TI - [Fee schedules and cost containment]. AB - Medical fee schedules are controversial. In this paper we examine the reasons that justify the imposition of fee schedules in the presence of a socially financed health insurance system, and examine the ways of constructing a medical fee schedule. The weakness of fee-for-service tariffs is that they do not allow a control of health care costs if the volume of services is unchecked. Current solutions to this problem--audit of doctors' average cost per case, freeze on new medical practices, or the insurers' discretion in choosing the doctors they reimburse--have multiple drawbacks. Alternatives to fee-for-service payment--such as flat fees, or payment based on the quality of medical services--are discussed. PMID- 19999313 TI - [Quality and saving: do they run together?]. AB - In the healthcare debate, it is often stated that better quality leads to savings. Quality systems lead to additional costs for setting up, running and external evaluations. In addition, suppression of implicit rationing leads to additional costs. On the other hand, they lead to savings by procedures simplification, improvement of patients' health state and quicker integration of new collaborators. It is then logical to imagine that financial incentives could improve quality. First evidences of pay for performances initiatives show a positive impact but also some limitations. Quality and savings are linked together and require all our attention. PMID- 19999314 TI - [Retroperitoneal fibrosis: a rare cause of postrenal kidney failure, no to be missed]. AB - We report the observation of a fifty years old man, admitted in the emergency room for bilateral lumbar pain and hyperkaliemic metabolic acidosis, and postrenal kidney failure induced by bilateral hydronephrosis. Radiographic exploration and histologic studies of biopsy confirmed an idiopathic retroperitoneal fibrosis that clinically and biologicaly responded to three seances of hemodialysis, and insertion in each uretere of one double J stent, and long term corticotherapy. The retroperitoneal fibrosis is a little common inflammatory disease, characterized by the development of a fibrous mass around the retroperitoneal structures. His diagnostic means evolved. On the other hand, his treatment was the object of no checked controlled and randomized trial. This article proposes an updating of the knowledge on this subject. PMID- 19999315 TI - [Are the guidelines the standards we have to follow]. AB - National or international medical societies yearly produce many recommendations i.e., guidelines about patient care, diagnostic or therapeutic procedures or preventive measures. These publications are established according to a level of evidence, based on randomised controlled trials or case studies. The strength of evidence reflects the presence of a good agreement or disagreement between the published trials. The clinician when confronted to these recommendations find them too rigid, poorly adapted to individual patients and depriving him from autonomy. However the use of well established guidelines plays a proven role in improvement of the quality and effectiveness of patient care. Primary care physicians should recognise the good guidelines and follow them to improve their daily practice. PMID- 19999316 TI - [Referring to an unusual case: pulmonary affection and rheumatoid arthritis]. AB - We report the case of a 60 year female patient suffering from rheumatoid arthritis for the last 25 years, under TNF-blocker and leflunomide, affected by a recurrent pneumothorax with several subpleural nodules, basal bronchiectasis and apical bullous emphysema. The patient was administered several treatments: aspiration, talc pleurodesis, surgical pleurodesis, pleurodesis induced by tetracycline and autologous blood. To allow the pleural inflammatory reaction necessary to the success of the pleurodesis, we had to interrupt the treatment by TNF-blocker and leflunomide. We then witnessed a partial pleurodesis with persistence of a pneumothorax. The medical situation is improving with disappearance of dyspnea. PMID- 19999317 TI - [Cerbral salt wasting syndrome versus SIADH]. AB - In the context of cerebral diseases the two main mechanisms responsible for non iatrogenic causes of hyponatremia are cerebral salt wasting syndrome (CSW) and inappropriate secretion of antidiuretic hormone (SIADH). Distinction between these two syndromes is difficult and is based on the assessment of the patient's volume status. In case of CSW, the volume status is low and the treatment is fluid and sodium replacement. In case of SIADH the volume status is normal or slightly expanded and the treatment is fluid restriction. To avoid centropontine myelinolysis, the speed of correction should not exceed 8 to 10 mmol/L over a 24 hour period. This article will describe practical tools to differentiate CSW from SIADH and therapeutic strategies useful in daily clinical practice. PMID- 19999318 TI - [Control of economizing by santesuisse.in medical offices: what practical message?]. PMID- 19999319 TI - [The evolution according to Darwin, in the era of robotics: hazard, need and intelligent design]. PMID- 19999320 TI - [Global budget that does not tell its name]. PMID- 19999321 TI - [The experts and Big Pharma, the minister and his guru]. PMID- 19999322 TI - [Red wines and fruits of the sea]. PMID- 19999323 TI - [Castration and the sex criminals (2)]. PMID- 19999324 TI - [HIV/AIDS: the new therapeutic strategies remain a priority]. PMID- 19999325 TI - [Late and unpopular vaccination]. PMID- 19999326 TI - An unusual cause of pelvic pain: struma ovarii. AB - A 22 year old female patient presented with a three month history of pelvic discomfort and dysmenorrhoea. A pelvic MRI demonstrated a large mass measuring 10 x 6 cm in size. On T1 and T2 weighted images the mass was noted to be predominantly cystic. The mass also contained peripheral solid components as well as a large fat fluid level. Following excision, histological examination revealed a lesion with mature teratomatous elements but with a significant component (> 50%) being composed of mature thyroid tissue. A 2cm area within the lesion had the morphological pattern of a classical papillary thyroid carcinoma. A diagnosis of struma ovarii was made. Struma ovarii is a rare ovarian neoplasm generally arising in a teratoma and accounts for less than 1% all ovarian neoplasms. Here we present the above case of struma ovarii in a young patient and discuss the radiological characteristics of the disease. PMID- 19999327 TI - Tuberculous arthritis of the finger: a forgotten disease. AB - We present a case of a 64-year-old man from Belgian origin with a chronic painful swelling of the third finger. A chronic arthritis from atypical germ was suspected at the third metacarpo-phalangeal joint on the plain radiography, ultrasonography and MR examination. A few months later, he developed thoracic back pain. Plain radiographs, a Technetium-99m bone scan and MR examination of the thoracic spine revealed a spondylodiscitis of the dorsal spine. A tuberculous origin was found on the biopsy of the affected dorsal vertebral bodies D9 and D10 as suspected on the first imaging examinations of the finger. Besides the patient was found to be HIV-positive. This case shows that it is important to think of musculoskeletal tuberculosis particularly in high-risk persons because if left untreated, it can be devastating. PMID- 19999328 TI - Malignant peritoneal mesothelioma: contribution and limitation of imaging for its diagnosis. AB - Malignant mesothelioma of the peritoneum is a rare primary peritoneal tumor with a pore prognosis. The overall prevalence is one to two cases per million. The tumor arises from mesothelial cells in the pleura or, in this case, peritoneum. Peritoneal malignant mesotheliomas account for only 10 to 20% of all malignant mesotheliomas. Exposure to asbestos fibers is a known risk factor for the development of mesothelial tumors. Although imaging features can be suggestive of malignant peritoneal mesothelioma, histologic examination of tissue with specific immunohistologic markers are mandatory for diagnosis. PMID- 19999329 TI - The black eyebrow sign in orbital blowout fracture. AB - Orbital blowout fractures are frequently associated with intraorbital emphysema. This may be seen on a frontal skull radiograph as a lucent area at the superior side of the orbit, simulating a black eyebrow. Although computed tomography remains the primary imaging modality for assessing patients with clinically obvious blowout fracture, conventional radiography is often the first imaging study performed in cases of orbital trauma. By consequensce, knowledge of the black eyebrow sign is very useful especially to more unexperienced radiologists working at the emergency department. PMID- 19999330 TI - Asymptomatic pneumatosis intestinalis in AIDS. AB - Pneumatosis intestinalis is an unusual entity in which subserosal or submucosal collections of gas occur in the bowel wall. We present a case of asymptomatic linear pneumatosis intestinalis and free abdominal air in a 67-year-old man with acquired immunodeficiency syndrome (AIDS) who improved spontaneously following conservative management. Pneumatosis intestinalis is a late-stage phenomenon in patients with AIDS that characteristically involves the cecum, ascending and transverse colon or the rectum with sparing of the descending colon and the sigmoid. For these patients, non-operative management is appropriate. PMID- 19999331 TI - Metastatic testicular germ cell tumor presenting with abdominal pain: CT and MRI findings. AB - Testicular tumors are the most common solid tumors in young adult males. Patients with testicular tumor often present with painless scrotal mass. Rarely, symptoms are related to metastases. We describe the case of a patient presenting with abdominal pain due to retroperitoneal lymphadenopathy. We describe the magnetic resonance imaging findings of lumbar spine osseous metastasis. PMID- 19999332 TI - Ultrasound appearances of Implanon implanted contraceptive devices. AB - Subdermal contraceptive devices represent a popular choice of contraception. Whilst often removed without the use of imaging, circumstances exist where imaging is required. Ultrasound is the modality of choice. The optimal technique and typical sonographic appearances are detailed in this article. PMID- 19999333 TI - Misleading appearance of atlantoaxial diastasis in Down syndrome: os odontoideum. PMID- 19999334 TI - MRI features of sacrococcygeal chordoma. PMID- 19999335 TI - Hot cross bun sign in a patient with SCA-2. PMID- 19999336 TI - Traumatic dissection of the extracranial vertebral artery. PMID- 19999337 TI - Patent foramen ovale and cryptogenic stroke. PMID- 19999338 TI - Multiscale image processing and antiscatter grids in digital radiography. AB - Scatter radiation is a source of noise and results in decreased signal-to-noise ratio and thus decreased image quality in digital radiography. We determined subjectively whether a digitally processed image made without a grid would be of similar quality to an image made with a grid but without image processing. Additionally the effects of exposure dose and of a using a grid with digital radiography on overall image quality were studied. Thoracic and abdominal radiographs of five dogs of various sizes were made. Four acquisition techniques were included (1) with a grid, standard exposure dose, digital image processing; (2) without a grid, standard exposure dose, digital image processing; (3) without a grid, half the exposure dose, digital image processing; and (4) with a grid, standard exposure dose, no digital image processing (to mimic a film-screen radiograph). Full-size radiographs as well as magnified images of specific anatomic regions were generated. Nine reviewers rated the overall image quality subjectively using a five-point scale. All digitally processed radiographs had higher overall scores than nondigitally processed radiographs regardless of patient size, exposure dose, or use of a grid. The images made at half the exposure dose had a slightly lower quality than those made at full dose, but this was only statistically significant in magnified images. Using a grid with digital image processing led to a slight but statistically significant increase in overall quality when compared with digitally processed images made without a grid but whether this increase in quality is clinically significant is unknown. PMID- 19999339 TI - Comparison of flat-panel digital to conventional film-screen radiography in detection of experimentally created lesions of the equine third metacarpal bone. AB - Radiographic diagnosis of equine bone disease using digital radiography is prevalent in veterinary practice. However, the diagnostic quality of digital vs. conventional radiography has not been compared systematically. We hypothesized that digital radiography would be superior to film-screen radiography for detection of subtle lesions of the equine third metacarpal bone. Twenty-four third metacarpal bones were collected from horses euthanized for reasons other than orthopedic disease. Bones were dissected free of soft tissue and computed tomography was performed to ensure that no osseous abnormalities were present. Subtle osseous lesions were produced in the dorsal cortex of the third metacarpal bones, and the bones were radiographed in a soft tissue phantom using indirect digital and conventional radiography at standard exposures. Digital radiographs were printed onto film. Three Diplomates of the American College of Veterinary Radiology evaluated the radiographs for the presence or absence of a lesion. Receiver operator characteristic curves were constructed, and the area under these curves were compared to assess the ability of the digital and film-screen radiographic systems to detect lesions. The area under the ROC curves for film screen and digital radiography were 0.87 and 0.90, respectively (P = 0.59). We concluded that the digital radiographic system was comparable to the film-screen system for detection of subtle lesions of the equine third metacarpal bone. PMID- 19999340 TI - Radiographic assessment of pulmonary fluid clearance in healthy neonatal foals. AB - We characterized the kinetics of postnatal equine lung using sequential thoracic radiography. The aim was to establish the earliest time when normal foals have clear, radiolucent lung fields, and to characterize the pattern of this clearance. Both right-to-left and left-to-right thoracic radiographs were acquired in lateral recumbency at peak inspiration within the first 30 min after birth and thereafter at 1, 2, 3, 4, 6, 8, 12, 24, 48, and 72 h. Radiographs were interpreted by three observers. The overall assessment of radiographic lung clearance was followed by the evaluation of individual lung quadrants to document changes in pulmonary radiographic patterns over time. We concluded that thoracic images in a healthy foal older than 4 h should be characterized by clear lungfields and that after this time distinctions between physiologic and pathologic conditions can be made. The ventral lung cleared first, presumably due to the greater flexibility of the thoracic wall in this anatomic region. PMID- 19999341 TI - An innovative technique for displaying three dimensional radiographic anatomy of synovial structures in the equine distal limb. AB - Radiography is the most commonly applied imaging modality in equine practice and forms an essential part of the diagnostic work-up of lame horses. Radiographic signs of musculoskeletal pathology are frequently localized at sites of soft tissue attachment, which are often not clearly visible on radiographs. Different lesions carry different prognoses and require a variety of treatments, and a good knowledge of the position of the synovial structures in the distal limb of the horse is essential for practitioners in the interpretation of radiographs. This study describes a new technique for creating three-dimensional (3D) models of the synovial structures and superimposing them onto radiographs for the purpose of teaching radiographic anatomy. A set of standard radiographs was acquired of the metacarpophalangeal and the distal interphalangeal joints of a fresh cadaver leg while the leg was positioned in a material-testing machine to mimic the weight bearing horse. Computed tomography of the same regions was performed after injection of negative contrast medium into the joints. 3D reconstructions of the joints were created using grayscale thresholding and polynomial surface meshing in Mimics. The resulting 3D reconstructions were superimposed on top of the radiographs using Adobe Photoshop CS3 Extended, thus allowing the visualization of the joint anatomy in relation to the bone on all projections. The main advantage of this technique is that it allows synovial structures to be visualized on radiographs where they are normally indistinct, which will serve as a teaching aid for anatomy. PMID- 19999342 TI - Magnetic resonance anatomy of the proximal metacarpal region of the horse described from images acquired from low- and high-field magnets. AB - While low-field magnetic resonance (MR) images can provide useful information in the investigation of proximal metacarpal region pain, an in-depth knowledge of anatomy and comparison with more detailed high-field images are essential to understand the meaning of different signal intensities within tissues. This anatomic description is based on low-field and high-field MR examination of 30 cadaver metacarpal regions of mature horses with no history of carpal or proximal metacarpal pain. Normal MR anatomy is described and is illustrated by high-field and low-field MR images in transverse, sagittal and dorsal planes. Normal anatomic variations of soft tissue and osseous structures are discussed. Differences between the signal intensity and definition of tissues on high-field and low-field MR images and in different pulse sequences are highlighted. Several structures could be evaluated in both high-field and low-field images that cannot easily be imaged using radiography and ultrasonography, including the abaxial margins of the suspensory ligament, the interosseous ligaments between the metacarpal bones and the carpometacarpal ligaments. Structures that have previously not been described in detail were also identified. PMID- 19999343 TI - Improved identification of the palmar fibrocartilage of the navicular bone with saline magnetic resonance bursography. AB - Fibrocartilage degeneration is the earliest pathologic finding in navicular disease but remains difficult to detect, even with magnetic resonance (MR) imaging. We hypothesized that injection of the navicular bursa with saline would improve accuracy of MR imaging evaluation of palmar fibrocartilage. Thoracic limbs were collected from 11 horses within 6 h of death. Imaging was performed with a 1.5 T magnet using sagittal 2D proton density and transverse 3D FLASH sequences with fat saturation. For the purpose of determining sensitivity and specificity of the MR images, fibrocartilage was classified as normal or abnormal, based on combination of the findings of gross and microscopic pathology. Thickness of fibrocartilage was measured on histologic sections and corresponding transverse FLASH MR images before and after injection of saline. A paired Student's t-test was used for comparison of measurements. Partial thickness fibrocartilage loss was present in 6 of 22 limbs. Sensitivity of precontrast MR images for detection of lesions was 100% while specificity was 6%. Saline MR arthrography resulted in both sensitivity and specificity of 100% based on consensus review. Mean histologic fibrocartilage thickness was 0.75 +/- 0.12 mm. Mean fibrocartilage thickness on precontrast transverse FLASH images was 0.93 +/- 0.065 and 0.73 +/- 0.09 mm on postsaline images. The histologic cartilage thickness was signficantly different from that in precontrast images (P<0.001) but not in images acquired after saline injection (P = 0.716). Based on our results, and using pulse sequences as described herein, navicular fibrocartilage can only be evaluated reliably for the presence of partial thickness lesions after intrabursal injection of saline. PMID- 19999344 TI - Magnetic resonance imaging of the canine brain at 7 T. AB - The purpose of this study was to describe relevant canine brain structures as seen on T2-weighted images following magnetic resonance (MR) imaging at 7 T and to compare the results with imaging at 1.5 T. Imaging was performed on five healthy laboratory beagle dogs using 1.5 and 7 T clinical scanners. At 1.5 T, spin echo images were acquired, while gradient echo images were acquired at 3 T. Image quality and conspicuity of anatomic structures were evaluated qualitatively by direct comparison of the images obtained from the two different magnetic fields. The signal-to-nose ratio (SNR) and contrast-to-noise ratio (CNR) were calculated and compared between 1.5 and 7 T. The T2-weighted images at 7 T provided good spatial and contrast resolution for the identification of clinically relevant brain anatomy; these images provided better delineation and conspicuity of the brain stem and cerebellar structures, which were difficult to unequivocally identify at 1.5 T. However, frontal and parietal lobe and the trigeminal nerve were difficult to identify at 7 T due to susceptibility artifact. The SNR and CNR of the images at 7 T were significantly increased up to 318% and 715% compared with the 1.5 T images. If some disadvantages of 7 T imaging, such as susceptibility artifacts, technical difficulties, and high cost, can be improved, 7 T clinical MR imaging could provide a good experimental and diagnostic tool for the evaluation of canine brain disorders. PMID- 19999345 TI - Computed tomography bronchial lumen to pulmonary artery diameter ratio in dogs without clinical pulmonary disease. AB - Bronchiectasis is diagnosed in humans using multiple computed tomography (CT) criteria, the most important being dilatation of the bronchi. The most widely used criterion for detection of bronchial dilatation is a bronchial lumen to pulmonary artery diameter (bronchoarterial [BA]) ratio >1.0. No studies have been performed to determine the BA ratio in normal dogs. Thoracic CT images of 24 dogs without clinical pulmonary disease were reviewed. The BA ratio of the lobar bronchi of the left cranial (cranial and caudal parts), right cranial, right middle, left caudal, and right caudal lung lobes was measured. The mean of the mean BA ratio for all dogs was 1.45 +/- 0.21 (99% confidence interval [CI] = 1.34 1.56). The mean of the mean BA ratio as determined by lung lobe was 1.45 +/- 0.04 (99% CI = 1.41-1.49). The range of individual BA ratios was 0.8-2.0. There was no significant difference in mean BA ratios as a function of lung lobe (P = 0.60). The BA ratio in these clinically normal dogs was consistent and may be a useful tool in evaluating for bronchiectasis on CT images. BA ratios >2.0 were not identified in this population, suggesting a threshold to differentiate normal from abnormal bronchi. PMID- 19999346 TI - Contrast-enhanced computed tomography as a preoperative indicator of vascular invasion from adrenal masses in dogs. AB - The purpose of this study was to determine if contrast-enhanced computed tomography (CT) accurately reflected vascular luminal invasion by adrenal masses in dogs. Medical records of 15 dogs with 17 adrenal masses that underwent preoperative CT imaging were reviewed. Presence or absence of vascular invasion was confirmed by surgery or necropsy/histology. Vascular invasion was identified correctly using contrast-enhanced CT in 11/12 dogs. The sensitivity and specificity of contrast-enhanced CT for vascular invasion compared with surgery or necropsy was 92% and 100%, respectively. The positive predictive value and negative predictive value of contrast-enhanced CT was 100% and 90%, respectively. The accuracy was 95%. In one dog, invasion of the phrenicoabdominal vein was not identified on CT images. Six of eight masses with vascular invasion where a histologic diagnosis was obtained were malignant. Four of four pheochromocytomas invaded adjacent vasculature. Contrast-enhanced CT provided accurate preoperative assessment of adrenal masses. Vascular invasion by adrenal masses in this study occurred by way of the lumen of the phrenicoabdominal vein with subsequent intraluminal extension into other veins, rather than by erosion through vessel walls. PMID- 19999347 TI - The effect of positioning on the radiographic appearance of caudodorsal mediastinal masses in the dog. AB - In this prospective study, the effect of thoracic positioning on the visibility and size of caudal esophageal masses caused by spirocercosis was investigated. Dorsoventral (DV), ventrodorsal (VD) as well as left lateral recumbent (LLR) and right lateral recumbent (RLR) thoracic radiographs of 28 dogs, diagnosed endoscopically with spirocercosis, were evaluated. The radiographic findings were compared with those of esophageal endoscopy. Masses were seen equally well in left vs. right recumbency as well as in DV vs. VD positions but in DV/VD views 86% of masses were detected whereas in lateral views only 50% of masses were identified. In spirocercosis-endemic areas DV and RLR views are advised as they also allow for better visualization of descending aorta aneurysms and to avoid interpreting the potentially normally visible esophagus in LLR in large dogs as a mass. PMID- 19999348 TI - Incomplete ossification of the atlas in dogs with cervical signs. AB - Osseous defects affecting the atlas were identified in computed tomography and magnetic resonance images of five dogs with cervical signs including pain, ataxia, tetraparesis, or tetraplegia. Osseous defects corresponded to normal positions of sutures between the halves of the neural arch and the intercentrum, and were compatible with incomplete ossification. Alignment between the portions of the atlas appeared relatively normal in four dogs. In these dogs the bone edges were smooth and rounded with a superficial layer of relatively compact cortical bone. Displacement compatible with unstable fracture was evident in one dog. Concurrent atlantoaxial subluxation, with dorsal displacement of the axis relative to the atlas, was evident in four dogs. Three dogs received surgical treatment and two dogs were treated conservatively. All dogs improved clinically. Incomplete ossification of the atlas, which may be associated with atlantoaxial subluxation, should be considered in the differential diagnosis of dogs with clinical signs localized to the cranial cervical region. PMID- 19999349 TI - Imaging diagnosis--acute necrotizing myopathy in a dog. PMID- 19999350 TI - Imaging diagnosis--intra-abdominal lipoma. PMID- 19999351 TI - Imaging diagnosis--magnetic resonance imaging of an organizing abscess secondary to a retrobulbar grass awn. PMID- 19999352 TI - A technique for ultrasound-guided paravertebral brachial plexus injections in dogs. AB - Using cadaveric dogs, we established the ultrasonographic landmarks for performing paravertebral injections around the brachial plexus nerve roots in the dog, and assessed the accuracy and regional spread of the aliquots. A mixture of methylene blue dye and an iodinated contrast medium was used as the injectate. A 0.3 ml volume was used to assess accuracy and a 3.0 ml volume was used to assess regional spread. Accuracy and regional spread were assessed from computed tomography (CT) images acquired after injection by measuring the distance from each foramen to the nearest edge of contrast medium, and the dimensions of spread of contrast medium in each anatomic plane, respectively. The mean distance of small volume injections from each foramen was 0.9 cm (standard deviation [SD] 0.56 cm). The mean spread of contrast medium for the small volume injections measured 1.7 cm (SD 0.6 cm) cranial-to-caudal, 1.2 cm (SD 0.4 cm) dorsal-to ventral and 7.4 cm (SD 0.4 cm) medial-to-lateral. The mean spread of contrast medium for the combined three large volume injections measured 7.4 cm (SD 1.7 cm) cranial-to-caudal, 3.1 cm (SD 0.8 cm) medial-to-lateral, and 2.8 cm (SD 0.5 cm) dorsal-to-ventral. After the CT studies, staining of each nerve root and any other regional structure was assessed grossly. Based on our results, ultrasound can be used to guide injections around the nerve roots of the brachial plexus in dogs. PMID- 19999353 TI - Regional brain perfusion in epileptic dogs evaluated by technetium-99m-ethyl cysteinate dimer SPECT. AB - We evaluated the feasibility of interictal single photon emission computed tomography (SPECT) to detect alterations in regional cerebral blood flow and neuronal activity in dogs with idiopathic epilepsy. Twelve dogs with idiopathic epilepsy underwent interictal technetium-99m-ethyl cysteinate dimer SPECT of the brain. Different cortical regions of interest (ROIs), 1 ROI at the cerebellum and 1 ROI at the subcortical area were evaluated by semiquantitative analysis and compared with a control group (18 dogs). Significant hypoperfusion (P = 0.02) was present in the subcortical area of epileptic dogs. This hypoperfusion was not associated with seizure frequency, age at onset of seizures, duration of epilepsy, or time since the last seizure. Interictal SPECT did not reveal cortical or cerebellar perfusion alterations. The subcortical area may play an important role in the pathophysiology of canine idiopathic epilepsy. PMID- 19999354 TI - Use of 3'-deoxy-3'-[18F]fluorothymidine PET/CT for evaluating response to cytotoxic chemotherapy in dogs with non-Hodgkin's lymphoma. AB - Imaging and measurement of proliferation with computed tomography (CT) and positron emission tomography (PET) provide a noninvasive method for improved staging and monitoring of response to cancer treatment. We evaluated prospectively the proliferation marker 3'-deoxy-3'[18F] fluorothymidine (FLT) in the context of FLT-PET/CT for detection of early response, confirmation of posttreatment response, and prediction of relapse in dogs with non-Hodgkin's lymphoma. Nine dogs with non-Hodgkin's lymphoma who were scheduled to receive five cycles of an investigational cytotoxic chemotherapy agent were included. All dogs received baseline FLT-PET/CT imaging immediately before chemotherapy. Intent was to repeat imaging with FLT-PET/CT at various time points: group 1 (n = 4), 5 days after initiation of chemotherapy and 3 weeks following the last chemotherapy administration; group 2 (n = 5), before the fourth cycle of chemotherapy and 3 weeks following the last administration. Two dogs in group 2 did not undergo repeat PET/CT. Body mass standardized uptake values (SUV) for FLT were calculated for each dog. Eight dogs had initially increased FLT uptake (mean SUVmax = 9.8 [2.6-22.3]). Mean SUV decreased significantly for the seven dogs that underwent follow-up PET/CT following chemotherapy (mean SUVmax = 3.5 [1.1-7.9], P<0.016). Increased uptake preceded clinical and cytological evidence of relapse in two dogs. Ki-67 immunohistochemistry confirmed decreased proliferation corresponding to decreased SUV in three canine lymph node samples. FLT-PET/CT functional and anatomical imaging shows promise for the evaluation of response to cytotoxic chemotherapy in dogs with non-Hodgkin's lymphoma and for predicting relapse before standard clinical and clinicopathologic confirmation. PMID- 19999355 TI - Radiotherapy and surgery for feline soft tissue sarcoma. AB - Medical records for 79 cats with soft tissue sarcomas treated with preoperative or postoperative curative intent radiation therapy between August 1994 and February 2004 were reviewed. The purpose was to assess the effectiveness of preoperative and postoperative radiation therapy, and to determine the association of patient and radiation treatment variables with survival. Gender, age, weight, anatomic tumor site, packed cell volume (PCV), computerized vs. manual treatment planning, radiation field length, preoperative vs. postoperative irradiation, total radiation dose, and biologically effective dose (BED) were assessed as prognostic factors for survival. Fifty-six of 79 (71%) of cats were anemic within 2 weeks before or during radiation treatment. The median survival was 520 days for all cats, with a 1-year survival rate of 61.6%, and a 2-year survival rate of 41.6%. Only timing of radiation therapy relative to surgery and presence of a moderate or severe anemia were significantly related to survival. The median survival was 310 days for cats treated with preoperative radiation therapy, and 705 days for cats treated with postoperative radiation therapy (P = 0.03). The median survival was 308 days for cats with a PCV<25%, and 760 days for cats with a PCV > or = 25% (P = 0.017). Radiation therapy in combination with surgery results in relatively long-term survival in cats with soft tissue sarcomas. Anemia is common in cats undergoing radiation therapy for soft tissue sarcomas, and is associated with decreased survival. PMID- 19999356 TI - Use of force plate analysis to evaluate the efficacy of external beam radiation to alleviate osteosarcoma pain. AB - A standard of therapy for osteosarcoma includes amputation with or without adjuvant chemotherapy. There is a subset of dogs with osteosarcoma that are unsuitable for amputation. We evaluated kinetic variables in dogs with appendicular osteosarcoma treated with a single 8 Gy dose of radiation. Eighteen pet dogs with appendicular osteosarcoma received one 8 Gy fraction of palliative radiation on day 0. Force plate measurements and clinical assessments were made on days 0, 7, 14, and 21. Peak vertical forces (Fz) were recorded for each limb and a symmetric index (SI) was calculated. There were no significant changes in kinetic parameters after one 8 Gy dose of radiation therapy. Nine of these 18 dogs exhibited increased limb function at day 21 based on force plate analysis. Significant factors affecting Fz included gender and tumor location. There was a significant correlation between Fz and response to therapy based on SI at day 21. SI seems to be useful to objectively assess response in this mixed population of dogs. One 8 Gy fraction of radiation therapy alone did not reduce lameness associated with appendicular osteosarcoma, but a subset of dogs did have improved limb function after a single dose. PMID- 19999357 TI - 3D conformal radiation therapy for palliative treatment of canine nasal tumors. AB - We evaluated the response of 38 dogs treated with a coarsely fractionated, palliative radiation protocol based on CT-based 3D treatment planning. Dogs with histologically confirmed malignant nasal tumors were studied. Treatment prescriptions consisted of 3-4 x 8 Gy, 4-5 x 6 Gy, or 10 x 3 Gy fractions. Selected patient and tumor factors were evaluated for an effect on outcome. Resolution of clinical signs was reported after irradiation in all dogs. Acute toxicities were mild and short lived. Thirty-seven of 38 dogs died or were euthanized due to tumor-related disease. Overall median progression-free interval (PFI) was 10 months. Tumor stage affected response, with modified stage 1 patients having a median PFI 21.3 months vs. a median PFI of 8.5 months for modified stage 2 patients (P = 0.0006). Modified stage was the only factor significantly related to outcome. Based on these findings, a palliative radiation prescription based on computerized treatment planning may be justified in some canine nasal tumor patients. PMID- 19999358 TI - A case for increased research on the pediatric upper airway. PMID- 19999360 TI - Endoscopic resection of juvenile nasopharyngeal angiofibroma. AB - OBJECTIVES: A 2-surgeon technique has been proposed that allows resection of juvenile nasopharyngeal angiofibroma (JNA) with extension into the infratemporal fossa by utilizing a septal incision for passage of a retracting instrument from the opposite nostril. This technique, however, does not overcome the problem of limited space within the nasal cavity for the tumor to be retracted. Therefore, the tumor has to be divided to allow for its removal. We are proposing a different 2-surgeon technique as an alternative operative technique for the resection of JNA. METHODS: A new technique of endoscopic resection of JNA involves a transseptal posterior perforation. This perforation allows retraction of the tumor into the opposite nasal cavity by the second surgeon. The retraction of the tumor creates space for its resection. RESULTS: Nineteen patients (all male) underwent this 2-surgeon technique for resection of JNA. The follow-up period ranged up to 9 years, and no recurrence was recorded. CONCLUSIONS: Longer term follow-up is needed to assess recurrence rate and morbidity with this technique. However, in our small series, the 2-surgeon technique via posterior septal perforation was associated with low morbidity and recurrence rates. PMID- 19999359 TI - Vocal exercise versus voice rest following botulinum toxin injections: a randomized crossover trial. AB - OBJECTIVES: The intensity of muscle activity immediately following intramuscular botulinum toxin injection may affect the clinical efficacy of the injection. We tested this effect in patients who underwent botulinum toxin injections for adductor spasmodic dysphonia. METHODS: Patients were studied over 3 to 5 injection cycles. Cycle 1 was the baseline control; cycle 2 was randomized between a 1-hour reading aloud task ("exercise") and a 24-hour period of complete voice rest. For cycle 3, the patient completed the task not performed in cycle 2. Patients who were willing to continue for cycles 4 and 5 repeated the experiment at one half the injection dosage. Efficacy was determined with a battery of voice recordings and clinical outcomes instruments administered via telephone at 2- to 4-week intervals. The primary outcome measure was the result of the Voice-Related Quality of Life (VRQOL) instrument. RESULTS: Nine patients (8 women, 1 man) with a mean age of 60.8 years (range, 42 to 76 years) completed at least 3 injection cycles. The VRQOL results were significantly higher for cycles that followed the exercise task. The patients reported subjectively that these were some of the best injection cycles they had ever experienced. Some achieved equivalent results with the half-dose injection plus exercise. The VRQOL results after voice rest cycles were not significantly different from the patients' baseline cycles. CONCLUSIONS: These results support the conclusion that a period of intense vocalization immediately following laryngeal botulinum toxin injections improves the efficacy of the injection. Possible mechanisms are proposed. PMID- 19999361 TI - Change of external auditory canal pH in acute otitis externa. AB - OBJECTIVES: We investigated (1) the correlation between the degree of acute otitis externa (AOE) and a change of pH and (2) the recovery of pH after acidification compared to an antibiotic otic solution in AOE. A change of pH in the external auditory canal (EAC) is very important for the pathogenesis of otitis externa. Therefore, not only an antibiotic otic solution, but also acidification, is known to be a good treatment for AOE. However, pH has only been investigated in chronic otitis externa, and not in AOE. METHODS: This was a prospective randomized control study. Forty adult patients (56 ears) with AOE and 40 normal control subjects (80 ears) participated in this study. The severity of disease was graded as mild, moderate, or severe. The pH of each EAC was then measured. The patients were randomly assigned into 2 groups: one for vinegar irrigation and the other for topical antibiotics. The pH of the diseased ears was measured at 1 and 2 weeks after the treatment. RESULTS: The mean (+/-SD) pH for the normal control subjects was 5.4 +/- 0.48, and the EAC lost its acidity proportionately to the degree of disease (p < 0.05). For both the vinegar irrigation and topical antibiotic groups, the acidity was restored dramatically (p < 0.05) at 1 and 2 weeks for the moderate and severe grades of otitis, but not for the mild grade. There was no difference in recovery between the 2 groups. CONCLUSIONS: The EAC lost its acidity proportionately to the degree of disease in AOE. Vinegar irrigation and topical antibiotics were equally effective for restoration of pH. PMID- 19999362 TI - Expression and distribution of hyaluronic acid and CD44 in unphonated human vocal fold mucosa. AB - OBJECTIVES: The tension caused by phonation (vocal fold vibration) is hypothesized to stimulate vocal fold stellate cells (VFSCs) in the maculae flavae (MFe) to accelerate production of extracellular matrices. The distribution of hyaluronic acid (HA) and expression of CD44 (a cell surface receptor for HA) were examined in human vocal fold mucosae (VFMe) that had remained unphonated since birth. METHODS: Five specimens of VFMe (3 adults, 2 children) that had remained unphonated since birth were investigated with Alcian blue staining, hyaluronidase digestion, and immunohistochemistry for CD44. RESULTS: The VFMe containing MFe were hypoplastic and rudimentary. The VFMe did not have a vocal ligament, Reinke's space, or a layered structure, and the lamina propria appeared as a uniform structure. In the children, HA was distributed in the VFMe containing MFe. In the adults, HA had decreased in the VFMe containing MFe. In both groups, the VFSCs in the MFe and the fibroblasts in the lamina propria expressed little CD44. CONCLUSIONS: This study supports the hypothesis that the tensions caused by vocal fold vibration stimulate the VFSCs in the MFe to accelerate production of extracellular matrices and form the layered structure. Phonation after birth is one of the important factors in the growth and development of the human VFMe. PMID- 19999363 TI - Hearing loss progression and contralateral involvement in children with unilateral sensorineural hearing loss. AB - OBJECTIVES: We undertook this study to determine the rate of hearing loss progression in the affected ear of children with unilateral sensorineural hearing loss and without an enlarged vestibular aqueduct, and the rate of new-onset hearing loss in the contralateral ear. METHODS: We searched the database at our pediatric tertiary care center to identify patients who met the inclusion criteria, examining demographic variables, audiometric data, and presumptive causes. RESULTS: We identified 198 patients. At presentation, they showed slight left-sided and male predominances. Of 142 patients who had sufficient audiometric follow-up for us to evaluate progression, 21% showed ipsilateral progression and 10.6% developed new-onset hearing loss in the contralateral ear. Isolated high frequency loss was identified in 11 patients (5.6%), 8 of whom had sufficient follow-up for us to identify progression. Two showed progression; 4 others with progression in the ipsilateral ear developed new-onset high-frequency loss in the contralateral ear. Temporal bone anomalies were identified in 26 children (13%), and these children were more likely to have profound hearing loss than were those without temporal bone anomalies (46% versus 23%). CONCLUSIONS: The findings suggest that unilateral sensorineural hearing loss may not always be a unilateral process, but that it may be the initial manifestation of bilateral auditory dysfunction. PMID- 19999364 TI - Midline cervical cleft: a rare congenital anomaly. AB - OBJECTIVES: A midline cervical cleft (MCC) is a rare congenital anomaly due to failure of fusion of the first and second branchial arches during embryogenesis. It may present as a midline defect of the anterior neck skin with a skin projection or sinus, or as a subcutaneous fibrous cord. This report evaluates the clinical features and surgical management of an MCC. METHODS: We analyzed a series of 4 patients with an MCC successfully treated at Great Ormond Street Hospital for Children in London. RESULTS: Three male patients and 1 female patient between 4 and 11 months of age were found to have an MCC. Each patient presented with an erythematous, fibrous band of tissue extending between the chin and the suprasternal notch. Treatment comprised surgical excision of the lesion and Z-plasty repair. We present the embryology, common clinical presentation, investigations, differential diagnosis, and histology, along with a literature review, of this uncommon malformation of the anterior neck. CONCLUSIONS: An MCC is a differential diagnosis to consider when assessing a child with a midline cervical lesion. Early surgical excision with Z-plasty repair of the soft tissue defect is the treatment of choice to prevent long-term complications. PMID- 19999365 TI - Respiration-related laryngeal electromyography in children with bilateral vocal fold paralysis. AB - We present 2 case reports to demonstrate the relationship between laryngeal muscle activity and respiration in children with bilateral vocal fold paralysis (BVFP) by simultaneous laryngeal electromyography (EMG) with recording of chest wall movement and intercostal muscle EMG. Laryngeal EMG was performed together with recording of chest wall movement in a 55-day-old girl who was undergoing tracheostomy for idiopathic congenital BVFP. Normal phasic activity was observed, i.e., the thyroarytenoid (TA) muscle was active during expiration and the posterior cricoarytenoid (PCA) muscle during inspiration, suggesting a good prognosis for recovery. The child was decannulated at 11 months. Laryngeal EMG together with recording of chest wall movement and intercostal EMG in a 5-year old girl who was tracheostomy-dependent following tracheoesophageal fistula repair due to BVFP showed phasic activity during expiration for both the TA and PCA muscles, indicating aberrant regeneration of the PCA motor nerve. The timing of laryngeal muscle activity with respiration in the assessment of pediatric congenital BVFP is essential to demonstrate the presence of normal or abnormal medullary respiratory neuronal input to laryngeal motoneurons. In cases in which BVFP is due to recurrent laryngeal nerve injury, respiration-related laryngeal EMG will identify aberrant regeneration. Laryngeal EMG should be combined with intercostal muscle EMG in the evaluation of children with significant vocal fold dyfunction of either central or peripheral origin. PMID- 19999366 TI - Bioengineered trachea with fibroblasts in a rabbit model. AB - OBJECTIVES: Although our group has had mostly successful results with clinical application of a tracheal prosthesis, delayed epithelial regeneration remains a problem. In our previous studies using rats, it was demonstrated that tracheal fibroblasts accelerated proliferation and differentiation of the tracheal epithelium in vitro and in vivo. The purpose of this study was to evaluate the effects of fibroblasts on epithelial regeneration in larger tracheal defects in rabbits. METHODS: We developed a bioengineered scaffold, the luminal surface of which was coated with fibroblasts. This scaffold was implanted into tracheal defects in 12 rabbits (bioengineered group), and scaffolds without fibroblasts were implanted in 12 rabbits (control group). The regenerated epithelium was histologically examined by light microscopy, scanning electron microscopy, and immunohistochemical studies. RESULTS: In the bioengineered group, a stratified squamous epithelium was observed on the surface 7 days after transplantation. However, in the control group, the scaffolds were exposed. Fourteen days after implantation, a columnar ciliated epithelium was observed in the bioengineered group. The average thickness of the regenerated epithelium in the bioengineered group was significantly greater than that in the control group. CONCLUSIONS: This study indicated that fibroblasts had a stimulatory effect that hastened regeneration of the epithelium in large tracheal defects. PMID- 19999367 TI - Atelocollagen sponge as a stem cell implantation scaffold for the treatment of scarred vocal folds. AB - OBJECTIVES: Treatment of vocal fold scarring remains a therapeutic challenge. Our group previously reported the efficacy of treating injured vocal folds by implantation of bone marrow-derived stromal cells containing mesenchymal stem cells. Appropriate scaffolding is necessary for the stem cell implant to achieve optimal results. Terudermis is an atelocollagen sponge derived from calf dermis. It has large pores that permit cellular entry and is degraded in vivo. These characteristics suggest that this material may be a good candidate for use as scaffolding for implantation of cells. The present in vitro study investigated the feasibility of using Terudermis as such a scaffold. METHODS: Bone marrow derived stromal cells were obtained from GFP (green fluorescent protein) mouse femurs. The cells were seeded into Terudermis and incubated for 5 days. Their survival, proliferation, and expression of extracellular matrix were examined. RESULTS: Bone marrow-derived stromal cells adhered to Terudermis and underwent significant proliferation. Immunohistochemical examination demonstrated that adherent cells were positive for expression of vimentin, desmin, fibronectin, and fspl and negative for beta III tubulin. These findings indicate that these cells were mesodermal cells and attached to the atelocollagen fibers biologically. CONCLUSIONS: The data suggest that Terudermis may have potential as stem cell implantation scaffolding for the treatment of scarred vocal folds. PMID- 19999368 TI - Predictors of aspiration pneumonia following radiotherapy for head and neck cancer. AB - OBJECTIVES: Aspiration following radiotherapy for head and neck cancer (HNC) is a common event, but not all patients with aspiration will develop pneumonia. Our aim was to identify predictors of pneumonia in patients with aspiration following radiotherapy for HNC. METHODS: We performed a retrospective study of 52 patients referred for modified videofluoroscopic barium swallow (MVBS) testing at our institution from 2003 to 2007 in order to identify clinical variables associated with the diagnosis of aspiration pneumonia. RESULTS: Independent risk factors for the development of pneumonia were tracheobronchial aspiration on MVBS testing (odds ratio [OR], 5.0; 95% confidence interval [CI], 1.2 to 20.5; p = 0.025), malnutrition (OR, 4.4; 95% CI, 1.3 to 14.7; p = 0.018), and smoking history (OR, 1.04 per pack-year; 95% CI, 1.01 to 1.07; p = 0.011). Through logistic regression analysis, we developed a bivariate predictive model with a sensitivity of 58%, a specificity of 90%, a positive predictive value of 79%, and a negative predictive value of 77% for the development of aspiration pneumonia in our patient population. CONCLUSIONS: Depth of aspiration on MVBS testing, malnutrition, and smoking history were strongly associated with the development of aspiration pneumonia in our patient population. The use of clinical variables to determine the risk of aspiration pneumonia is feasible and may help identify high-risk patients. PMID- 19999370 TI - Workforce deployment--a critical organizational competency. AB - Staff scheduling has historically been embedded within hospital operations, often defined by each new manager of a unit or program, and notably absent from the organization's practice and standards infrastructure and accountabilities of the executive team. Silvestro and Silvestro contend that "there is a need to recognize that hospital performance relies critically on the competence and effectiveness of roster planning activities, and that these activities are therefore of strategic importance." This article highlights the importance of including staff scheduling--or workforce deployment--in health care organizations' long-term strategic solutions to cope with the deepening workforce shortage (which is likely to hit harder than ever as the economy begins to recover). Viewing workforce deployment as a key organizational competency is a critical success factor for health care in the next decade, and the Workforce Deployment Maturity Model is discussed as a framework to enable organizations to measure their current capabilities, identify priorities and set goals for increasing organizational competency using a methodical and deliberate approach. PMID- 19999369 TI - Anatomy-based algorithms for detecting oral cancer using reflectance and fluorescence spectroscopy. AB - OBJECTIVES: We used reflectance and fluorescence spectroscopy to noninvasively and quantitatively distinguish benign from dysplastic/malignant oral lesions. We designed diagnostic algorithms to account for differences in the spectral properties among anatomic sites (gingiva, buccal mucosa, etc). METHODS: In vivo reflectance and fluorescence spectra were collected from 71 patients with oral lesions. The tissue was then biopsied and the specimen evaluated by histopathology. Quantitative parameters related to tissue morphology and biochemistry were extracted from the spectra. Diagnostic algorithms specific for combinations of sites with similar spectral properties were developed. RESULTS: Discrimination of benign from dysplastic/malignant lesions was most successful when algorithms were designed for individual sites (area under the receiver operator characteristic curve [ROC-AUC],0.75 for the lateral surface of the tongue) and was least accurate when all sites were combined (ROC-AUC, 0.60). The combination of sites with similar spectral properties (floor of mouth and lateral surface of the tongue) yielded an ROC-AUC of 0.71. CONCLUSIONS: Accurate spectroscopic detection of oral disease must account for spectral variations among anatomic sites. Anatomy-based algorithms for single sites or combinations of sites demonstrated good diagnostic performance in distinguishing benign lesions from dysplastic/malignant lesions and consistently performed better than algorithms developed for all sites combined. PMID- 19999371 TI - An innovative approach to goal setting and reporting in a complex organization. AB - Managing multiple priorities and achieving sustainable outcomes is a challenge for leaders across health care organizations. One academic health science centre set about to address this challenge through the development of an innovative annual goal and objectives process that aligned every unit/department around organizational priorities. The results have enabled exceptional outcomes for individuals, teams, patients/families and the organization as a whole. PMID- 19999372 TI - From a project to transformation: how "going against the flow" led to improved access and patient flow in an academic hospital. AB - A results-driven approach to optimizing patient flow, grounded on quality improvement, change management and organizational learning principles, is described. Tactics included collaborative governance, performance management, rapid process improvements and implementation toolkits. Results included an 83.1% decrease in emergent volumes waiting for greater than 24 hours and a 49.1% improvement in emergency department length of stay for admitted patients. There were no adverse outcomes on other key indicators. Sustainability remains the challenge but early results are encouraging. PMID- 19999373 TI - Towards a collaborative model of care. AB - The current crisis in Canada's health care system calls for transformational change in the way we deliver care. The Collaborative Model of Care is not a new concept, but has not been implemented in Canadian acute care hospitals until recently. Toronto East General Hospital developed and piloted a collaborative care model on three acute units and initial results are promising in terms of improved patient safety, patient satisfaction, job satisfaction and improved use of resources. PMID- 19999374 TI - Adverse events in older patients admitted to acute care: a preliminary cost description. AB - The financial costs associated with Adverse Events (AEs) for older patients (> or = 65 years) in Canadian hospitals are unknown. The objective of this paper is to describe and compare costs between patients who experienced an AE and those who did not during an acute hospital admission to a tertiary care facility. Patients with an AE had twice the hospital length of stay (20.2 versus 9.8 days, p < 0.00001), resulting in 1,400 extra days at a cost of approximately $7,500/patient. PMID- 19999375 TI - [Research on zoonoses: central element of the "One Health" initiative]. AB - The idea of "Comparative Medicine" was founded by distinguished scientists in the 1900s, in which context the name of Rudolf Ludwig Carl Virchow always comes into the limelight. This discipline was practiced thoroughly with the study of every infective disease transmitted naturally from vertebrates to man, namely the zoonoses. The following article will highlight the concept of "Comparative Medicine" with the example of tuberculosis research carried out during the second half of the 19th century. After the discipline took an interesting turn toward increasing specialization in the field, particularly after the Second World War, infection research in veterinary and human medicine developed and progressed in different directions. Since the 1990s, this development has contradicted the awareness, that a great number of diseases can be successfully combated through interdisciplinary efforts only, which has now led to the concept of "One health". In view of this, functional molecular epidemiology will play an invaluable role in the future in recognizing associations between host and pathogen-genotypes, particularly in research pertaining to emerging and re-emerging infections, which include the zoonoses. Zoonotic research is the crux of such a concept; hence, this article will portray some of the developments in Germany over the last few years, by virtue of which the "One health" dogma brings zoonotic research to life. PMID- 19999376 TI - Epidemiology and pathogenesis of enterohaemorrhagic Escherichia coil. AB - Enterohaemorrhagic Escherichia coli (EHEC) cause diarrhoea, haemorrhagic colitis and the haemolytic uraemic syndrome (HUS). EHEC cause sporadic, epidemic, and occasionally fatal infections in Germany and worldwide. The most probable natural reservoirs of EHEC are domestic cattle (in which these organisms cause no disease), but wild animals also excrete EHEC asymptomatically. Humans (chiefly children) acquire EHEC from contact with animals (e. g. in petting zoos) or indirectly, by consuming food or water contaminated with these pathogens. Acid resistant EHEC survive the acidic human stomach, colonize the intestines, and produce Shiga toxins. Current models suggest that Shiga toxins preferentially bind to microvascular endothelial cells of the renal glomeruli and the brain and inhibit protein synthesis, or possibly injure eukaryotic cells via other mechanisms. Resulting microangiopathy, therefore, forms the pathological basis of HUS. Additionally, non-Shiga toxin effectors produced by EHEC, including the LEE encoded proteins, EHEC-haemolysin, cytolethal distending toxin (CDT), the serine protease EspP, subtilase cytotoxin, and iron-acquisition molecules are also postulated to play roles in virulence. Our evolving understanding of EHEC pathogenesis now compels us to test the hypothesis that HUS is a multifactorial consequence of EHEC infection, that involves not only Shiga toxins, but a repertoire of EHEC virulence factors that synergistically cause profound vascular and organ injury. PMID- 19999377 TI - Bat rabies--a Gordian knot? AB - Although classical rabies is one of the earliest identified and best studied infectious diseases, there is still limited knowledge about lyssaviruses and their major natural hosts, bats. Focussing on bat rabies in Europe caused by European bat lyssaviruses 1 (EBLV-1) and 2, for instance the association of EBLV 1 to Eptesicus bats and EBLV-2 to Myotis daubentonii and M. dasycneme together with an apparent clustering of cases is one question still to be answered. Furthermore, the question whether EBLVs are less virulent or bats less susceptible is the key to the understanding of the disease. Accumulating evidence from experimental studies and field observations, however, has resulted in contradicting hypotheses. Serological surveys, using tools developed for classical rabies, are often used for bat rabies surveillance. However, such surveys are hampered by the lack of validated methods applicable for bat sera. Bats seem to play a prominent role as reservoir for viral pathogens and the unique biology of bats especially the immune response may contribute to this. Considering all known aspects, bat rabies seems to form a yet unsolvable entanglement, reminiscent of the ancient tale of the Gordian knot. In this manuscript we will not be able to untangle this knot, but we hope to offer some suggestions of where to start. PMID- 19999378 TI - [From mice to men: the threat of hantavirus infections]. AB - Hantaviruses are naturally hosted by small mammals. Virus transmission to humans can result in the development of Hemorrhagic Fever with Renal Syndrome (Asia, Europe) or Hantavirus Cardiopulmonary Syndrome (The Americas). Certain hantaviruses can cause life-treatening infections with case fatality indices up to about 50%. Different virus types have evolved which use different species of small mammals (mainly but not exclusively rodent species) as natural reservoirs. It is still unknown why these different hantaviruses exhibit a different virulence when transmitted to humans as accidental hosts. This paper reviews the nature and molecular epidemiology of hantaviruses as well as strategies to identify new virus types. PMID- 19999379 TI - [Bird populations--hatching grounds of pandemic influenza viruses?]. AB - Aquatic wild birds constitute the natural reservoir of influenza A viruses. Under appropriate selection pressure these viruses display a remarkable genetic flexibility which is based on their high mutation rate. At least 16 subtypes of the viral hemagglutinin glycoprotein (HA) und 9 of the viral neuraminidase (NA) are distinguishable as a result of natural evolution. Due to the segmentation of the viral genome, genetic reassortments and, thus, a theoretically unrestricted combination of HA and NA subtypes is possible. Transspecies-specific transmissions of viruses from the natural reservoirs to mammals (horses, swine, humans) led to the establishment of several influenza virus lineages which are circulating stably and independently from the previous reservoir. Introduction of viruses featuring new HA/NA combinations into immunologically naive populations can cause rapid and widespread, even pandemic, dissemination of such viruses. The (highly pathogenic avian influenza, HPAI). These mutants constitute the causative agents of classical fowl plague. HPAI viruses of subtypes H5 and H7 generally share a polybasic endoproteolytical cleavage site in the HA protein which allows for systemic replication in the avian host. Subtype H5 and H7 viruses which are maintained in the natural reservoir hosts, in contrast, are of low pathogenicity (LPAI) and harbour monobasic HA cleavage sites restricting their tissue tropism to epithelia of the respiratory and gastronintestinal tracts. This context forms the basis of all legal measures targeting the monitoring, prevention and eradication of HPAIV and LPAIV of subtypes H5 and H7 in poultry. Results of such measures in Germany in 2008 are presented and discussed. PMID- 19999380 TI - Character-based DNA barcoding: a superior tool for species classification. AB - In zoonosis research only correct assigned host-agent-vector associations can lead to success. If most biological species on Earth, from agent to host and from procaryotes to vertebrates, are still undetected, the development of a reliable and universal diversity detection tool becomes a conditio sine qua non. In this context, in breathtaking speed, modern molecular-genetic techniques have become acknowledged tools for the classification of life forms at all taxonomic levels. While previous DNA-barcoding techniques were criticised for several reasons (Moritz and Cicero, 2004; Rubinoff et al., 2006a, b; Rubinoff, 2006; Rubinoff and Haines, 2006) a new approach, the so called CAOS-barcoding (Character Attribute Organisation System), avoids most of the weak points. Traditional DNA-barcoding approaches are based on distances, i. e. they use genetic distances and tree construction algorithms for the classification of species or lineages. The definition of limit values is enforced and prohibits a discrete or clear assignment. In comparison, the new character-based barcoding (CAOS-barcoding; DeSalle et al., 2005; DeSalle, 2006; Rach et al., 2008) works with discrete single characters and character combinations which permits a clear, unambiguous classification. In Hannover (Germany) we are optimising this system and developing a semiautomatic high-throughput procedure for hosts, agents and vectors being studied within the Zoonosis Centre of the "Stiftung Tierarztliche Hochschule Hannover". Our primary research is concentrated on insects, the most successful and species-rich animal group on Earth (every fourth animal is a bug). One subgroup, the winged insects (Pterygota), represents the outstanding majority of all zoonosis relevant animal vectors. PMID- 19999381 TI - [What makes an insect a vector?]. AB - Blood-feeding insects transmit numerous viruses, bacteria, protozoans and helminths to vertebrates. The developmental cycles of the microorganisms in their vectors and the mechanisms of transmission are generally extremely complex and the result of a long-lasting coevolution of vector and vectored pathogen based on mutual adaptation. The conditions necessary for an insect to become a vector are multiple but require an innate vector competence as a genetic basis. Next to the vector competence plenty of entomological, ecological and pathogen-related factors are decisive, given the availability of infection sources. The various modes of pathogen transmission by vectors are connected to the developmental routes of the microorganisms in their vectors. In particular, pathogens transmitted by saliva encounter a lot of cellular and acellular barriers during their migration from the insect's midgut through the hemocele into the salivary fluid, including components of the insect's immune system. With regard to intracellular development, receptor-mediated invasion mechanisms are of relevance. As an environmental factor, the temperature has a paramount impact on the vectorial roles of hematophagous insects. Not only has it a considerable influence on the duration of a pathogen's development in its vector (extrinsic incubation period) but it can render putatively vector-incompetent insects to vectors ("leaky gut" phenomenon). Equally crucial are behavioural aspects of both the insect and the pathogen such as blood host preferences, seasonal appearance and circadian biting activity on the vector's side and diurnal/nocturnal periodicity on the pathogen's side which facilitate a contact in the first place. PMID- 19999382 TI - [What comes after bluetongue--Europe as target for exotic arboviruses]. AB - Many factors led to the situation in which we fear the appearance of yet another exotic microbe simply demonstrating our vulnerability to new invaders and the shrinking of the globe. Beside the tremendous increase in international travel and trade, climatic changes are a crucial factor for exotic arboviruses to establish endemic transmission cycles in Europe. Certain temperatures, humidity and--at least for mosquitoes--suitable water bodies are required for the respective species to develop and become abundant. Increasing temperatures reduce the overall time of the mosquito life cycle because the aquatic developmental stages are completed faster, and secondly, the time span required for the mosquito to be capable to transmit the acquired arbovirus to another susceptible host is shortened. Thus the chances for an exotic arbovirus to establish an autochtonous transmission cycle are increasing. Here, a review is provided on the known factors that determine vector competence for mosquitoes and a particular arbovirus. The comparison of the closely related West Nile and Usutu viruses in Europe and North America further demonstrates how different and unpredictable the epidemiology of such arboviruses can be. Time will answer the question on who is going to be the next arboviral intruder into Europe. But it is today that we can bring ourselves into a good position to conquer the next "alien" by setting up interdisciplinary networks to promote research on these "exotic" and thus neglected pathogens thereby enhancing our understanding on their transmission and the factors and mechanisms required for this. PMID- 19999383 TI - [Surveillance of wild birds for avian influenza A virus (AIV) in Bavaria in the years 2007 and 2008]. AB - A monitoring programme has been initiated in Bavaria to continuously control wild birds for the presence of avian Influenza A virus (AIV) and to monitor the possible occurrence and accumulation of notifiable AIV subtypes as an early warning system. In addition information about the regional, seasonal and species specific distribution of AIV could be obtained. Between July 2007 and December 2008 samples from 5864 wild birds of twelve different zoological orders had been collected (cloacal- and tracheal swab samples, droppings, and organs) and analysed. AIV genomes were detected in 3.7% of the 5864 wild birds by RT real time PCR. The subtype component H5 was identified in 52 samples (0.9%) and the N1 subtype component in 13 samples (0.2%), but never in combination with each other. The hemagglutinine subtype component H7 could not be detected. Most of the positive AIV genome results originated from samples in the district Swabia, which is situated in the central area of the south-west bird migration route across southern Germany and harbours favourable resting areas for migrating birds. Mallards (Anas platyrhynchos) were the most frequently sampled bird species and had the highest AIV infection rate of 6.4%, followed by Tufted ducks (Aythya fuligula) (AIV prevalence of 5.4%), Mute Swans (Cygnus olor) (1.6%), Coots (Fulica atra) (0.3%) and Greylag Goose (Anser anser) (0.1%). The detection rate of AIV in Bavarian wild birds showed a seasonal peak in autumn/winter. Ten virus isolates could be obtained after sample inoculation in embryonated hen's eggs. PMID- 19999384 TI - Bioequivalence, therapeutic equivalence and generic drugs. PMID- 19999385 TI - Geriatrics on the run: rationale, implementation, and preliminary findings of a Belgian internal liaison team. AB - This article describes the rationale, implementation, interventions and preliminary findings of a Belgian interdisciplinary internal liaison team in a 1470-bed teaching hospital. The motive to start the team was threefold: the ageing of the inhospital population, the conclusion that health care professionals working on non-geriatric wards often lack the necessary skills to deal with older patients' needs and Belgian law, obliging each general hospital to set up an internal liaison team. Our team aims at detecting geriatric patients at risk, assisting health care professionals in caring for older patients and sensitizing them regarding optimal geriatric care. The article explains the underlying philosophy and strategy for implementation, focusing on the concepts of reciprocity, flexibility and cooperation. The preliminary results are based on a process evaluation of 719 consultations carried out from November 2004 to November 2006, a time registration, and a Strengths, Weaknesses, Opportunities, and Threats analysis (SWOT). Although our data are preliminary and the implementation of the team was pragmatic rather than research driven, they provide insight into the development, implementation, functioning and interventions of a Belgian interdisciplinary internal liaison team. PMID- 19999386 TI - Economic evaluation of voriconazole versus caspofungin for the treatment of invasive aspergillosis in Belgium. AB - OBJECTIVE: To perform an economic evaluation of voriconazole versus caspofungin in first line treatment of invasive aspergillosis (IA). These 2 antifungal drugs have a more favorable toxicity profile than the conventional amphotericin B and have a lower cost than the expensive liposomal/lipid formulation of amphotericin B. No head-to-head comparative study was conducted with voriconazole and caspofungin in IA. Based on the clinical trials of both antifungals, a conservative approach of similar efficacy has been considered. METHODS: The analysis is based on a simplified cost-minimization model with results from the National Health system RIZIV/INAMI perspective (year 2008). Only limited direct costs were considered, namely the drug cost over the episode of treatment.Treatment duration and patients' weight were key parameters. Their values were obtained from the Belgian observational VORIBEL study (Pfizer data on file) for voriconazole treatment. Treatment duration for caspofungin was derived from the EORTC study where almost 50% of the patients were recruited in Belgian centres. Mean cost and incremental cost were calculated. Univariate sensitivity analyses were carried out on weight, treatment duration as well as on route of administration. RESULTS: In invasive aspergillosis, the weighted cost per episode of fungal infection was 11.996 pound with voriconazole treatment (voriconazole IV followed by oral voriconazole) and 13.657 pound with caspofungin treatment (intravenous caspofungin only). The incremental saving with voriconazole treatment was 1.661 pound per patient.The cost-saving results of voriconazole were confirmed with varying treatment duration within realistic range. 41% of the patients in the VORIBEL study were fully treated with oral formulation. For these patients a saving of 6.375 pound was achieved with the use of oral voriconazole. CONCLUSION: Voriconazole is a cost-saving option compared with caspofungin in the treatment of invasive aspergillosis. PMID- 19999387 TI - Health care provision in prisons: a review on European and international guidelines. AB - Health care provision in prisons is an area of increasing international concern. The spectrum of health problems which prisoners may bring to prison is wide and in many cases prevalence is greater than in the general population.The profound knowledge of delicate issues such as medical confidentiality and informed consent of the patient, with special refer to infectious disease, hunger strike and prison research is necessary in the prison working environment. In the following paragraphs the European and International conventions and guidelines on the matter are discussed. PMID- 19999388 TI - Generics and cost-effective prescribing in Belgium: does bioequivalence always translate in therapeutic equivalence? AB - As many other countries, Belgium has a policy to promote the use of generic pharmaceutical products. In order to protect consumers, these generic products must be demonstrated to be essentially similar to the previously approved product, typically an innovator product. The therapeutic equivalence of a generic and an innovator product is most commonly based on the demonstration of bioequivalence, i.e. clinically insignificant differences in the rate and extent of drug absorption usually assessed from pharmacokinetic measurements, in a normal and healthy population. This article reviews the bioequivalence requirements for generic products and examines whether bioequivalence always adequately substantiates therapeutic equivalence and interchangeability. Clinical practice has identified a number of drug classes for which generic substitution should be approached with caution. Current bioequivalence requirements are based on a measure of average bioequivalence. There are fears that use of this measure may be inappropriate in the case of a drug with a narrow therapeutic range or high intrasubject or intersubject variability. Under these circumstances, measures of individual and population bioequivalence are proposed to be more accurate than measures of average bioequivalence. Bioequivalence issues are discussed together more general concerns about generic drug substitution, such as differences in product and packaging appearance and differences in excipients. PMID- 19999389 TI - Generics: need for clinical concern? AB - The market of generic drugs is in continuous development in all countries, including Belgium. Their low cost explains their success in western and developed countries. However, clinical concerns have been raised when generics are used. Indeed, various studies suggest that generic substitution can be associated with reduced efficacy or (and) increased side-effects, particularly with drugs used in severe diseases or pathological states such as epilepsy, cardiac arrhythmia, prevention of graft-rejection, ... The generic drugs must have systemic bioavailability similar to that of the original drug. Thus, they have supposed similar therapeutic bioequivalence. However, similar pharmacokinetics does not imply identical therapeutic activity, particularly with drugs having narrow therapeutic indices such as anti-epileptics, anti-arrythmics ... In this case, switchability rather than prescribability may cause problems. Low pharmaceutical quality is more frequent when drugs are produced in certain countries, in some cases causing a real concern when activity and safety are considered. PMID- 19999390 TI - Short-term return visits of 'general internal medicine' patients to the emergency department: extent and risk factors. AB - OBJECTIVES: Although emergency department (ED) return visits are a significant problem universally, it has not been previously studied in our ED. The aim of this study was to determine the extent of the problem in our ED, to identify the relevant clinical predictor variables and to detect diagnostic errors. METHODS: A retrospective observational study of ED return visits by patients managed by the General Internal Medicine (GIM) service was performed. The study was performed over a one year period at a tertiary hospital ED. Data are reported as relative risk (RR) and 95% confidence interval (CI). RESULTS: There were a total of 51.210 ED visits during the study period. The total number of ED return visits within 72 hours was 1.124 (2,19%; 95% CI 2,07 to 2,32). The total number of ED patients managed by the GIM service was 9.511. The percentage of patients treated by the GIM service who returned to the ED within 72 hours was 1,48% (95% CI 1,25 to 1,74) when calculated for the whole group and 2,9 % (95% CI 2,46-3,41) for those discharged home from the ED (n = 4.860). The majority (82,98%) of ED return visits by patients discharged from the GIM service were unscheduled and related to their index presenting complaint. Abdominal pain was the commonest initial presenting symptom in the patients who returned to the ED after discharge. Patients with diarrhoea as the initial initial presenting symptom had the highest relative risk of an ED return visit (RR = 4.07). CONCLUSION: The percentage ED return visits by patients discharged from the ED by the GIM service is 1,48%. Patients presenting with diarrhoea as the initial presenting symptom have the highest relative risk of an early ED return visit. Our main practical conclusion is that patients with abdominal pain need to be re-examined carefully and instructed about potential evolution before discharge. PMID- 19999391 TI - JAK2 V617F mutation and PRV-1 overexpression: relevance in the diagnosis of polycythaemia vera and essential thrombocythaemia. AB - In the diagnosis of polycythaemia vera and essential thrombocythaemia, two molecular markers were described in the last decade: the overexpression of the PRV-1 gene and the V617F mutation in the JAK2 gene. In this study we assess their usefulness by comparing our test results with the available clinical data. We show that in the diagnosis of polycythaemia vera the JAK2 mutation screening is crucial, while testing for the PRV-1 overexpression is redundant. On the contrary, in the diagnosis of essential thrombocythaemia (ET), both JAK2 and PRV 1 show their usefulness. PMID- 19999392 TI - Concurrence of multiple myeloma and idiopathic erythrocytosis. AB - The association of erythrocytosis and multiple myeloma is rare. We encountered a 76-year-old male patient with erythrocytosis followed by the diagnosis of multiple myeloma 8 months later. Related laboratory examinations revealed absolute erythrocytosis, normal oxygen saturation and erythropoietin (EPO) levels, the absence of endogenous erythroid colony (EEC) and JAK2-V617F mutations. The diagnosis of idiopathic erythrocytosis (IE), instead of polycythemic vera (PV), was made. In the literature, about 20 cases of erythrocytosis associated with myeloma can be found. Based on elevated EPO levels, 2 of such cases should be considered secondary erythrocytosis while others are reported as PV. None of them is considered to be have idiopathic erythrocytosis. Our present case is the first one with multiple myeloma developing in a patient with the diagnosis well established by extensive laboratory workup. The pathogenic role of these two entities remains to be established. PMID- 19999393 TI - Prostate abscess following intravesical BCG therapy. PMID- 19999394 TI - Rupture of the ureter mimicking acute renal infarction: case report and review of the literature. AB - We describe a case of acute rupture of the left proximal ureter as a result of lithiasis. The patient presented with a clinical picture of renal infarction: unilateral flank pain, more than 500 red blood cells per microliter urine and increased serum LDH levels. Abdominal ultrasound as well as a CT scan showed no abnormalities. Only CT angiography of the kidneys was able to demonstrate an acute rupture of the left proximal ureter with extravasation of contrast. Intravenous pyelogram confirmed this rupture with leaking of the contrast. One day after admission the patient urinated a 2 mm large calciumoxalate-monohydrate stone, the likely cause of the rupture. The rupture healed spontaneously. A review of the literature is given. PMID- 19999395 TI - A 50-year-old man with severe hypercalcemia: a case report. AB - OBJECTIVE: We present this case to emphasize the importance of early diagnosis and treatment of an acute severe hypercalcemic syndrome due to primary hyperparathyroidism as a consequence of an undiagnosed adenoma of the parathyroid gland. CASE REPORT: A 50-year-old man presented at another hospital with non specific symptoms such as anorexia, nausea, vomiting, polyuria, dehydration, abdominal pain, weight loss, fatigue, muscular weakness, irritability and lethargy. Serum levels of calcium and parathyroid hormone (PTH) were markedly increased to 23.6 mg/dL (reference values 8.6-10.2 mg/dL) and > 1900 ng/L (reference values 14-72 ng/L) respectively. After initial treatment, the patient was transferred to the intensive care unit (ICU) of a tertiary care university hospital for further stabilization and treatment because the typical signs of hypercalcemia were not resolving. A parathyroid adenoma was diagnosed and a few days later a parathyroidectomy was performed. The postoperative course was uneventful and the patient could be discharged from the hospital in a good general condition. CONCLUSION: Acute primary hyperparathyroidism, also known as parathyroid storm or parathyroid crisis, is a rare but potentially fatal endocrine emergency if unrecognized and untreated. Appropriate diagnosis and immediate adequate management of hypercalcemia are important in reducing mortality. Nevertheless, mortality remains high, even with surgical treatment which is the cornerstone of the definitive therapy. PMID- 19999396 TI - Autoimmune haemolytic anaemia associated with a thymoma: case report and review of the literature. AB - A 67-year-old female presents with a small mass in the anterior mediastinum on chest computed tomography. A biopsy proves the mass to be a spindle-cell-type or type A thymoma. Subsequently the patient develops fever and severe Coombs positive haemolytic anaemia. She is initially treated with oral corticosteroids. Because of persistence of the haemolysis subsequent thymectomy is performed. Haemolysis disappears almost instantly and does not return after discontinuation of the oral corticosteroids. Review of the literature reveals only 17 other cases of thymoma-associated autoimmune haemolytic anaemia. PMID- 19999397 TI - Necrotizing fasciitis due to Streptococcus pneumoniae in a healthy 68-year old man: case report and review of the literature. AB - A case of pneumococcal necrotizing soft-tissue infection (NSTI) is described and 16 cases available in literature are reviewed. Pneumococcal NSTI seems to be the consequence of hematogeneous shedding of pneumococci, with frequent involvement of articulations. Furthermore, pneumococcal NSTI present like other NSTI, with a high mortality and the need for a combined surgical and medical therapy. PMID- 19999398 TI - Pulmonary metastatic calcification in adult T-cell lymphoma/leukemia. PMID- 19999399 TI - Pneumocephalus during cabergoline treatment of an invasive macroprolactinoma. PMID- 19999400 TI - No more free lunch. PMID- 19999401 TI - The silver lining. PMID- 19999402 TI - Trigger point management. PMID- 19999403 TI - Major depression disorder. PMID- 19999404 TI - Diabetic foot ulcers. PMID- 19999406 TI - Surgical staples for scalp laceration repair. A quick and easy solution. PMID- 19999405 TI - Harnessing the hot flash. More choices for midlife women. PMID- 19999407 TI - Hepatitis A vaccination in children: an overview. PMID- 19999408 TI - It's a great life if you don't weaken. Reflections on living and dying. PMID- 19999409 TI - Managing MRSA in retail health. PMID- 19999410 TI - MRSA risk in children. Widening scope of infection requires action. PMID- 19999411 TI - Expanding your role. Do you really want to? PMID- 19999412 TI - Return to sender. Convince specialists to respond to NPs. PMID- 19999413 TI - NPs meeting acute needs. PMID- 19999414 TI - Changing a broken system. Clinic design gives patients options. PMID- 19999415 TI - Men and Alzheimer's disease. Sex differences in risk. PMID- 19999416 TI - Evaluating chest X-rays. Use mnemonics to develop a systematic approach. PMID- 19999417 TI - Body shaping. Today's solutions offer more to patients. PMID- 19999418 TI - The treatment and prevention of head lice infestation. PMID- 19999419 TI - Dry skin. Tips for everyday care. PMID- 19999420 TI - Evaluation of the acute abdomen. Key issues in primary care settings. PMID- 19999421 TI - Abdominal pain in a retail setting. PMID- 19999422 TI - Nurse practitioners in acute care. PMID- 19999423 TI - The scope of acute care practice. Hammering out the details. PMID- 19999424 TI - NP hospitalists. A new niche for NPs. PMID- 19999425 TI - Beyond birth control. The health benefits of hormonal contraception. PMID- 19999426 TI - Toilet trouble. A stepwise process can spell success. PMID- 19999427 TI - Cross communication. Avoiding device interference in pacemakers and ICDs. PMID- 19999428 TI - A only your doctor can prescribe? It's time for a new script. PMID- 19999429 TI - Respect and the DNP. PMID- 19999430 TI - Economic stimulus. Can a bad economy be good for retail health? PMID- 19999432 TI - Genitourinary disorders in children. A review of less common presentations. PMID- 19999433 TI - Bridging the disconnect. Better asthma care in hospitals and homes. PMID- 19999434 TI - No fear here. Overcoming the stigma of urinary incontinence. PMID- 19999431 TI - Late-life issues. Noting elder abuse, deciding on advance directives. PMID- 19999435 TI - Evaluation of cough in adults. A review of evidence-based guidelines. PMID- 19999436 TI - Quick care tips. Evaluation of cough in children. PMID- 19999437 TI - Trigeminal neuralgia. A debilitating pain syndrome. PMID- 19999438 TI - Management of dog and cat bites. Act quickly for best outcomes. PMID- 19999439 TI - Phrases NPs hate to hear. Strategies for drive-you-crazy questions. PMID- 19999440 TI - My friend John. PMID- 19999441 TI - Buying a preowned practice. PMID- 19999442 TI - Obesity in men. PMID- 19999443 TI - Rejuvenating aging hands. PMID- 19999444 TI - Lumbar puncture. PMID- 19999445 TI - How to quit smoking. PMID- 19999446 TI - Treating chronic pelvic pain. Focus on pain triggers and neurogenic inflammation. PMID- 19999447 TI - Lifting a fork to heart health. Whole foods as a treatment intervention. PMID- 19999448 TI - Treating cough in children. In most cases, watch, wait and review. PMID- 19999449 TI - The sun's evil enterprise. Malignant melanoma. PMID- 19999450 TI - Slow motion. Management of constipation. PMID- 19999451 TI - Presenting effective inservice. PMID- 19999452 TI - Overcoming roadblocks. Stop whining, and get back on track. PMID- 19999453 TI - Visibility at a price. Is retail health where we want to be? PMID- 19999454 TI - Get clinical experience before NP school. PMID- 19999455 TI - Why retail health? NPs count the reasons. PMID- 19999456 TI - Signing a consulting physician. Benefits can outweigh hassles. PMID- 19999457 TI - Experience is the best teacher. Answers to common billing questions. PMID- 19999458 TI - Patient information. Glucose monitoring. A guide to checking your blood sugar. PMID- 19999459 TI - Always say 'yes'. Scharmaine Lawson-Baker, NP: entrepreneur of the year. PMID- 19999460 TI - One to watch: Michael Lawler, NP. One business leads to another. PMID- 19999461 TI - Outside the box: Helen Carcio, NP. Restoring dignity through dryness. PMID- 19999462 TI - Hypertension update. Getting patients to goal with newer agents. PMID- 19999463 TI - Condoms for women. New options protect and empower. PMID- 19999464 TI - Herpes zoster alert. Prevent shingles with vaccination and awareness. PMID- 19999465 TI - 2008 in review: more growing pains. Twinges as the NP role expands. PMID- 19999466 TI - Time management. Ten commandments for reshaping your schedule. PMID- 19999467 TI - Disruptive innovation. NPs are true health care reformers. PMID- 19999468 TI - The DNP in retail health. Can a degree impact an industry? PMID- 19999469 TI - Writing for the public. Parenting Web site promotes NPs. PMID- 19999470 TI - Inflammmatory bowel disease. Sex differences in treatment effects. PMID- 19999471 TI - Subungual hematoma. Two evacuation procedures. PMID- 19999472 TI - General computer security. Protect yourself from viruses and spies. PMID- 19999473 TI - Patient information. A cold or the flu. Which is it? PMID- 19999474 TI - Impaired to empowered. Toward a better understanding of ADHD in adults. PMID- 19999475 TI - The doctorate of nursing practice. Coming into focus. PMID- 19999476 TI - Influenza's widening scope. Dangers prompt broader vaccination. PMID- 19999477 TI - Infants from birth to three. Do you see what they see? PMID- 19999478 TI - Ovarian cancer. Clinical and genetic considerations. PMID- 19999479 TI - Resolutions we can keep. Changes for patients and NPs. PMID- 19999480 TI - The DNP. Will it create a second-class NP? PMID- 19999481 TI - Risky business. PMID- 19999482 TI - Turning a negative into a positive. Addressing opposition to retail health clinics. PMID- 19999483 TI - Family matters. Should you hire your relatives? PMID- 19999484 TI - Reduction of a dislocated shoulder. Passive and active methods, step by step. PMID- 19999485 TI - Health implications of obesity. Women face unique risks and consequences. PMID- 19999487 TI - Patient information. Examining your skin: an important step in melanoma cure. PMID- 19999486 TI - Keeping your technology current. Do you really need a new PC? PMID- 19999488 TI - Refresh and renew. Getting started in medical aesthetics. PMID- 19999489 TI - Varicose veins. More than a cosmetic nuisance. PMID- 19999490 TI - Gaining control over eczema. An overview of diagnosis, treatment and prevention. PMID- 19999491 TI - Closing the door on fertility. Sterilization methods for women. PMID- 19999492 TI - Reducing risk behaviors in adolescents. New tool offers rapid assessment. PMID- 19999493 TI - Obstructive sleep apnea and ischemic stroke. A murky but menacing connection. PMID- 19999494 TI - Trichomoniasis. An update on diagnostic tools and treatments. PMID- 19999495 TI - When change won't work. Where is your line in the sand? PMID- 19999496 TI - A long and agonizing wait. Did I dodge the bullet again? PMID- 19999497 TI - Hollywood lays an egg; NAPNAP scrambles it. PMID- 19999498 TI - Becoming uninvisible. Use NP-to-NP networking to promote the profession. PMID- 19999499 TI - Rash, muscle weakness and pain. Is it dermatomyositis? PMID- 19999500 TI - Guardians ad litem. How do they differ from legal guardians? PMID- 19999501 TI - How to make sure you get paid. Tales from the trenches. PMID- 19999502 TI - Patient information. Colic: don't worry, it's normal. Tips for riding out the storm. PMID- 19999503 TI - Autism in school-age chidren. A complex collage of development, behavior and communication. PMID- 19999504 TI - Polycystic ovary syndrome in adolescents. A hormonal barrage and metabolic upheaval. PMID- 19999505 TI - The nurse practitioner in assisted living communities. A role poised for growth. PMID- 19999506 TI - Weight loss for patients with metabolic syndrome. An overview of diet choices. PMID- 19999507 TI - Laceration repair with tissue adhesives. A solution for simple wounds. PMID- 19999508 TI - Patient education in the fast lane. Streamline, organize and update. PMID- 19999509 TI - Bringing life into focus. An overview of myasthenia gravis. PMID- 19999510 TI - Stay positive in a negative environment. Five steps that make sense. PMID- 19999511 TI - Being unavailable. Pay attention to your priorities. PMID- 19999512 TI - Coincidental hero. The public trusts NPs. PMID- 19999513 TI - Merry-K. Moos. Pioneer for preconception health. PMID- 19999514 TI - Multivitamin use among non-pregnant females of childbearing age in the Western North Carolina multivitamin distribution program. AB - BACKGROUND: Daily consumption of 400 mcg of folic acid prior to conception and throughout the first trimester of pregnancy reduces the risk of neural tube defects (NTDs) by 50%-80%. A daily multivitamin with folic acid can ensure that females receive the recommended amount of folic acid during childbearing years. OBJECTIVE: The purpose of this study was to determine if vitamin consumption is influenced by providing a free bottle of multivitamins to non-pregnant women of childbearing age during a face-to-face interaction with a health care provider in health departments. METHODS: An eight-question survey was given to a sample of women who had received a free bottle of multivitamins. Vitamin consumption behavior prior to the intervention was compared to current usage at the time of the survey. RESULTS: Twenty-five percent of all survey respondents reported taking a daily multivitamin or folic acid tablet before the intervention. Fifty three percent reported taking a daily multivitamin 8-10 months later, a greater than two-fold increase (PR=2.1). Latino women reported the greatest increase in daily multivitamin intake, from 21% to 70% (PR=3.3). LIMITATIONS: The results may be difficult to extrapolate to the general population as the survey population differs from the general population. Prior vitamin use was determined by patient recall. The intervention occurred simultaneously with a multifaceted, public folic acid campaign. CONCLUSIONS: Eight to ten months after receiving a free three-month supply of multivitamins during a face-to-face interaction with a health care provider, the number of participants reporting daily use increased significantly. PMID- 19999516 TI - Selecting an electronic medical record system for small physician practices. AB - BACKGROUND: While electronic medical record (EMR) systems have demonstrated the potential to improve quality of care and reduce medical errors, relatively few practitioners have implemented EMR systems. This article presents a case study that explores the process by which small physician practices may select an ambulatory EMR system. METHODS: We assessed the appropriate criteria small practices should use in selecting an EMR system and then evaluated a range of commercially available EMR systems according to cost, functionality, and interoperability with existing systems. RESULTS: The process for selecting an EMR system starts by creating a budget for start-up costs and monthly maintenance expenses. Next, a practice should evaluate its strategic objectives and current computer infrastructure. The group should then define the appropriate functionality requirements specific for their practice. Finally, a certified ambulatory EMR system that interfaces with existing office systems can be selected. LIMITATIONS: This case study explores the process of EMR selection for rural, solo physician practices. The ability to generalize the process described herein to broader types of physician practices, such as multi-specialty group practices or to those practices with larger budgets for EMR systems, may be limited. CONCLUSIONS: Multiple critical and often competing factors--including cost and interoperability with existing systems, as well as organizational goals and obstacles--influence the selection of an EMR system for small physician practices. However, by following a standardized process for selecting an EMR system, small physician practices will find EMR selection to be a relatively straightforward process. PMID- 19999515 TI - Patient characteristics associated with receipt of lifestyle behavior advice. AB - BACKGROUND: Lifestyle behaviors such as reducing weight if overweight or obese, reducing salt intake, exercising, reducing alcohol intake, quitting smoking, and eating a healthy diet are related to the prevention and control of chronic diseases. However the amount of lifestyle advice provided by clinicians has been declining over the last decade. METHODS: In 2002, a telephone survey was conducted to assess the quality of preventive care offered by health care providers. The study was a cross-sectional observational study of a randomly selected sample of 516 diverse individuals in Durham County, North Carolina. Information regarding age, sex, race, education, health conditions, and self reported receipt of lifestyle advice was examined in the study. RESULTS: The odds of receiving advice to engage in preventive lifestyle behaviors were significantly higher for those with a pre-existing diagnosis of diabetes or hypertension and for participants reporting poor health status. For example, the odds of receiving advice to control or lose weight was 8.32 (95% CI, 2.65, 26.75) among individuals reporting a diagnosis of diabetes. Similarly, the odds of reporting "receiving advice to reduce salt intake" was 6.97 (95% CI, 3.74, 13.00) among subjects reporting a diagnosis of hypertension. LIMITATIONS: The results are from a cross-sectional study of a sample of individuals in only one county. Additionally, the results are based on patient self-reported information, which could be subject to recall and social desirability bias. CONCLUSION: Patients with identified health problems were more likely than others to report being advised to adopt healthy lifestyle recommendations. Future research should examine methods to encourage health care providers to offer lifestyle advice to those without pre-existing illness. PMID- 19999517 TI - Market hazard, moral imperative: why we need health reform. PMID- 19999518 TI - Get health reform right, not quick. PMID- 19999519 TI - 2009 North Carolina Child Health Report Card. PMID- 19999520 TI - Introduction. Preconception care: building the foundation for healthy women, babies, and communities. PMID- 19999521 TI - Issue brief: preconception care: building the foundation for healthy women, babies, and communities. PMID- 19999522 TI - Speeding the progress to improved health and wellness for North Carolina's women. PMID- 19999523 TI - Recommendations for women of reproductive age. PMID- 19999524 TI - State budget's effect on women's health care. PMID- 19999525 TI - The unique role of family physicians in caring for women across the reproductive age span. PMID- 19999526 TI - The role of North Carolina Medicaid in women's health and wellness. PMID- 19999527 TI - The role of specialists in providing preconception health care and guidance to women with chronic medical conditions. PMID- 19999528 TI - Promoting healthy weight in women: what the physician can do to help. PMID- 19999529 TI - The problems no one wants to see: mental illness and substance abuse among women of reproductive age in North Carolina. PMID- 19999530 TI - Pills are not enough: the case for long-acting reversible contraceptives--and how North Carolinians can benefit. PMID- 19999531 TI - Preconceptional considerations and counseling for the infertile couple. PMID- 19999532 TI - Taking a lead role in preconception health promotion in eastern North Carolina. PMID- 19999533 TI - Highlights from the North Carolina Preconception Health Strategic Plan. PMID- 19999534 TI - Reaching women and health care providers with women's wellness messages: the North Carolina Folic Acid Campaign as a model. PMID- 19999535 TI - Improving the health of working women: aligning workplace structures to reflect the value of women's labor. PMID- 19999536 TI - Resources for women's health. PMID- 19999537 TI - Using Master Settlement Agreement funds to reduce prenatal/postpartum smoking in North Carolina: you quit, two quit. PMID- 19999538 TI - Body mass index among North Carolina women of reproductive age. PMID- 19999539 TI - Alamance County Health Department. PMID- 19999540 TI - [Evaluation of morphologically determined apoptotic index]. AB - Stimulated by the huge scientific interest in programmed cell death, the sophisticated but expensive methods of apoptosis detection, occasionally even in various stages of the apoptotic process, have been developed in the last two decades. These methods are quite demanding and time consuming, e.g., TUNEL, flow cytometry and its modifications, immunohistochemistry, hybridization in situ, etc. Irrespective of considerable technical improvements, these methods are associated with a relatively high rate of false-positive and false-negative results. Reading off the results obtained by these methods is mostly left to investigators inexperienced in morphological analysis. It is well known that the basic morphological characteristics of apoptotic cells and apoptotic bodies have been described on the basis of classic light microscopy of histologic hemalaun eosin (HE) stained slides. As a rule, apoptotic cell has a shrunken, condensed, dark red cytoplasm and dark purple picnotic nucleus, and is surrounded by light, 'hollow' space (halo). Apoptotic bodies are fragments of the disintegrated apoptotic cell and each fragment of dense cytoplasm usually contains a fragment of disintegrated nucleus. It is known that, unlike necrosis, there is no inflammatory reaction in the vicinity of apoptotic cells and apoptotic bodies. During more than 25 years of research into apoptosis, the Zagreb Group for the Study of Apoptosis (Apoptosis Section, Department of Basic Medical Sciences, Academy of Medical Sciences of Croatia) have found it possible to determine the number of apoptotic cells and apoptotic bodies (apoptotic index) in daily routine on classic HE stained histologic slides by counting in 10 large fields under light microscope, following the methodology of mitotic index determination. Of course, the method is not absolutely accurate (the sophisticated and expensive methods mentioned above are not so either), but it is rapid and inexpensive. Thus, we believe that each pathologist should be professionally obliged to determine apoptotic index when assessing mitotic index. Expression of these indices as a mitotic-apoptotic ratio, e.g., 1:1, 1:2, 2:1, etc., may better reflect the growth potential of the examined, usually tumor tissue than determination of the mitotic index alone. PMID- 19999541 TI - [Genetic and molecular insights into apoptosis]. AB - Apoptosis, or programmed cell death, is a crucial mechanism for the survival of organisms and is functionally conserved in all higher eukaryotes. Molecular events in apoptosis are responsible for removal of damaged or infected cells from the cellular population, which links apoptosis to the cell cycle, replication and DNA repair. Moreover, apoptosis is one of the main mechanisms governing accurate embryonic development and maintenance of tissue homeostasis. In the invertebrate species Drosophila melanogaster and Caenorhabditis (C.) elegans apoptotic mechanisms are relatively simple, whereas in vertebrates they become more complex and include large groups of molecular components. There are two distinct apoptotic pathways in mammals. The first is called the "death receptor pathway" and is initiated by extracellular ligands such as TNFalpha, or FasL (Fas ligand)/CD95L, TWEAK and TRAIL. They bind to their receptors on the cellular surface, i.e., TNFR, Fas/CD95, DR3, DR4/DR5. The second pathway that also leads to the caspase activation is under the control of the Bcl-2 family of genes and proteins. Although C. elegans has only two members of Bcl-2 family, CED-9 and EGL 1, higher eukaryotes possess 30 homologs of this protein family. They can be categorized as those that promote cellular survival and those that exert proapoptotic action and induce programmed cell death. The final result of both apoptotic signaling pathways is the activation of cellular caspase. When considering a broader picture of apoptosis, these biological phenomena can today be interpreted in the new light as a basic mechanism of an organism life preserving. PMID- 19999542 TI - [Lysosomes and apoptosis]. AB - In 1955, Christian de Duve and his coworkers from the School of Medicine in Louvain, Belgium, named a group of cytoplasmic formations surrounded by lipoprotein membrane and containing acid hydrolase enzymes as lysosomes. Biochemical and cytochemical studies showed lysosomes to be found in animal and vegetable eukaryotic cells. Later on, lysosomes were found to be involved in the dynamics of lysosomal system, which consists of a number of various cytoplasmic formations such as primary and secondary lysosomes, endosomes, autophagosomes and postlysosomes. These formations are inter-connected by the mechanism of membrane integration, and in some instances by the cell membrane. Lysosomal system is involved in numerous physiological processes such as degradation of endogenous and exogenous macromolecules (proteins, lipids, polysaccharides and nucleic acids), cytoplasmic formations (mitochondria, peroxisomes, Golgi complex) that have performed their functions, tissue regression (post-lactation mammary gland), hormone secretion regulation (proinsulin to insulin), etc. On the other hand, lysosomal system is also involved in a number of pathologic processes like inflammation, allergic reactions, ischemia, hypoxia, as well as in lysosomal diseases (thesaurismoses), e.g., type II glycogenosis, fucosidosis, mucolipidosis III, etc. In 1974, Christian de Duve introduced the term lysosomotropism, denoting entry of the pharmacologically active, toxic and carcinogenic substances in the lysosomal system irrespective of their chemical nature and mechanism of input. Lysosomotropic substances may act in two ways: (1) causing impairments in the intralysosomal area, with toxic manifestations; and (2) modifying their membrane properties by increasing or decreasing membrane permeability, or by inducing labilizing or stabilizing effects. Damage to the lysosomal system occurs in the late stage of necrosis, while destabilization of lysosomal formations has been recorded in primary processes during apoptosis. PMID- 19999543 TI - [Psychoneurogenic factors and apoptosis]. AB - Immunity is regulated by its signal molecules, interleukins, as well as by cellular contacts; however, in addition to these 'local' mechanisms there is also neuroendocrine regulation of the immune system, at the level of the organism as a whole that attunes the immunoreaction up or down. The systemic regulation is accomplished by nervous and endocrine signals that translate and transmit inputs from emotional, cognitive and associative processes in the brain. At the cellular level, neuroendocrine signals join local networks to influence biochemical processes in the cytoplasm and nuclei of the lymphocytes, macrophages and other cells of the immune system. The neuroendocrine mediators glucocorticoids and opioid peptides may trigger apoptosis as a final mechanism after binding to specific cytoplasmic viz. membrane receptors and activating the Fas molecule at the cell membrane as the apoptosis promoter. Neuroendocrine mechanisms may also influence carcinogenesis, angiogenesis and tumor metastasis. PMID- 19999544 TI - [Apoptosis in toxicologic pathology]. AB - Toxicologic pathology provides expertise to the interpretation of the toxicity and safety of pharmaceuticals, biological agents, human and animal food aditives, environmental and industrial chemicals, and medical devices in animal studies. The histopathology findings are integrated with other study data (clinical and biochemistry data, autopsy) providing a comprehensive report on efficacy and safety of a chemical, device or material and the relationship of toxicity to exposure. Since its discovery, apoptosis emerged as a molecular control point in the regulation of physiological processes, toxic insults and diseases by means of a programmed cell death. Numerous factors include chemicals, oxidative stress, anoxia, and irradiation. Suppression, overexpression or mutation of a number of genes which orchestrate the apoptotic process are associated with disease. Also, the disbalance in apoptosis processes is documented in viral, autoimmune and neurodegenerative diseases, as well as in tumors. Research in the pharmacologic industry is driven toward developing new drugs for treatment schedules for these and other diseases. Toxicologic pathology findings of apoptosis should assist regulatory agencies in understanding the potential hazard or benefit of the tested substance (is the finding of apoptosis normal variation, spontaneous event or provoked by tested drug). Since the great majority of initial histopathological examinations made on toxicity studies and animal disease models are done on routine hematoxylin and eosin-stained slides, the morphology alone is sufficient for accurate and adequate identification of apoptosis. PMID- 19999545 TI - [Apoptosis during embryo development]. AB - The development of human embryo includes two essential processes, i.e., rapid mitotic activity of cells and gradual differentiation of tissues and organs. The latter process is very often characterized by extensive migration of cells from their site of origin to the site of definitive location, inductive action of the neighboring germ layers and programmed cell death (apoptosis). This paper describes examples of proliferative and apoptotic processes during the development of human embryo. The development of trilaminar germ disk, skin, gonads, central and peripheral nerve system as well as limbs provides instructive examples of how apoptosis regulates the development and differentiation of cells. PMID- 19999546 TI - [Apoptosis in tumors]. AB - The apoptotic mode of cell death is an active and defined process that plays an important role in the development of multicellular organisms and in the regulation and maintenance of the cell populations in tissues under physiologic and pathologic conditions. Defects in apoptosis can result in cancer. The apoptotic process is often described by use of apoptotic index. A high apoptotic index may predict good outcome in some tumor types and poor survival in patients with some other types of tumors. According to study results, the prognostic value of apoptosis might be tumor specific and its value as an independent prognostic index has yet to be established. PMID- 19999547 TI - [Apoptosis in pathologic prostatic processes]. AB - Numerous studies have demonstrated the apoptotic index to be significantly higher in benign prostatic lesions than in PIN and adenocarcinoma. Furthermore, a lower apoptotic index in epithelial cell populations has been shown to correlate with decreased immunoreactivity for prostate specific antigen that may be seen in higher Gleason grade tumors. It is suggested that the loss of function of some apoptotic regulators contributes to the development of PIN and prostatic adenocarcinoma. Of these, three signaling molecules involved in apoptotic functions ofTGF-beta have now been intensively investigated, including transmembrane receptor II (T betaRII), cell cycle inhibitor p27(Kp1) and Smad4, effectors of TGF-beta signaling pathway. Increased expression of p53 and decreased expression of maspin, a serine protease inhibitor, also play a major role in the apoptotic process. Changes in the expression of these markers may serve as a reliable criterion on making the diagnosis in biopsy material and may help choose an appropriate therapeutic approach. PMID- 19999548 TI - [Apoptosis, UV-radiation, precancerosis and skin tumors]. AB - Apoptosis plays an important role in regulating skin development and homeostasis, as well as carcinogenesis. Apoptosis balances epidermal proliferation to maintain epidermal thickness and may eliminate mutated, premalignant cells. Skin carcinomas are the most common cancers in Western population. The risk is closely related to UV exposure and the relation between exposure to UV radiation and development of skin cancer has been well established. Apoptosis and carcinogenesis in skin can be triggered by UV irradiation, and tumor suppressor gene p53 plays the most important role in these two processes. The p53 gene is involved in the cell cycle arrest and activation of apoptosis. It has been shown that p53 could protect skin cells from DNA-damage due to UVB exposure. UV-induced DNA damage activates the mechanisms for removal of DNA damage, delay in cell cycle progression, DNA repair, or apoptosis by transcriptional activation of p-53 related genes, such as p21 and bax. Several studies have shown that UVB also induces mutations in p53 gene.The induced mutations are unique (C toT and CC to TT transition) for UVB radiation and are not commonly induced by other carcinogens. Mutations in the p53 gene have been detected in 50% of all human cancers and in the majority of skin carcinomas. A high frequency of p53 mutation was reported in pre-malignant actinic keratosis lesions that is considered to be pre-squamous cell carcinoma, and in Bowen's disease that is considered to be in situ squamous cell carcinoma (SCC) of the skin. The majority of these mutations were characteristic UVB mutations and these findings suggested that p53 mutations might be involved in the malignant conversion of precancerous lesions to SCC. Several studies have demonstrated the continued and discontinued regimens of chronic UVB treatment to ultimately result in skin tumor development with 100% incidence, although the kinetics of tumor occurrence is delayed in the latter. Thus, these studies suggest that skin cancer development can be delayed but not stopped with further avoidance of UV exposure. More recent studies have shown that besides UVB, UVA component of solar irradiation could also be an important carcinogen but in the stem cell compartment of the skin. This knowledge will lead to development of new apoptosis-based therapeutics; several have recently been tested in SCC. It can be expected that new apoptosis-based drugs are to be introduced soon to the daily medical practice; however, prevention of excessive UVB exposure remains the main curative factor against skin cancer. PMID- 19999549 TI - [Cerebral ischemia and neuronal regeneration]. PMID- 19999550 TI - [Flow alteration treatment for giant aneurysm in the posterior cranial fossa]. PMID- 19999551 TI - [Intraoperative neuronavigation system without rigid pin fixation]. AB - OBJECT: Integration of neuronavigation into brain tumor surgery can improve the accuracy and precision of resection, but most neuronavigation systems require rigid pin fixation of the patient's head throughout the operation. We describe use of a noninvasive skull reference tool, which can replace rigid pin fixation in awake craniotomy and pediatric surgery, when standard pin fixation cannot be used. METHODS AND RESULTS: The skull reference tool (BrainLAB, Munich, Germany) is directly attached to the outer skull with a small self-tapping screw. Virtual scanning of the surface of the face and head achieves patient registration in seconds without requiring the use of fiducial markers or head fixation. This procedure improves patient comfort, cooperativeness, and compliance, resulting in better operating procedures. This skull reference tool is available for children younger than 3 years of age, in whom pin fixing is dangerous. Twelve patients underwent awake surgery and four pediatric patients younger than 3 years old underwent five procedures using this skull reference tool in our hospital between January 2005 and December 2008. The method of utilizing this skull reference tool was easy and sufficiently accurate in these patients. CONCLUSION: The use of this skull reference tool provides an accurate and comfortable method of frameless navigation in patients undergoing awake craniotomy and pediatric patients. PMID- 19999552 TI - [Radiation injury after stereotactic irradiaton: especially long-term follow-up benign of targets]. AB - OBJECTIVE: To analyses the result of linac radiosurgery (LRS) for the treatment of intracranial benign lesions and to assess possible factors related to complications. METHODS: The authors retrospectively reviewed 204 patients treated with LRS between May, 1993 and December, 2003. The study determined the correlation between radiosurgical complications including imaging changes after LRS and multiple factors such as radiosurgical parameters, location, volume and shape. We divided the patients into three groups by MRI imaging changes and clinical symptoms. Group 1 (Gr. 1): Imaging change only. Group 2 (Gr. 2): Imaging change with transient symptoms. Group 3 (Gr.3): Imaging change with permanent symptoms. RESULT: Ninty-three patient with AVM: Gr. 1, 8 cases (8.6%), Gr. 2, 1 case (1.1%), Gr. 3, 2 cases (2.1%). A significantly higher incidence of imaging change was noted in patients with arteriovenous malformation (AVM) volumes greater than 10 cc, irregular shape of the nidus and deep location. Fifty-eight patients with vestibulan schwannoma (VS): Gr. 1, 6 cases (10.3%), Gr. 2, 1 case (1.8%), Gr. 3, 2 cases (3.4%). Imaging changes were seen mostly in cases with tumor volume greater than 5 cc. Fifty-three patients with meningioma: Gr. 1, 4 cases (7.5%), Gr.2, 2 cases (3.7%), Gr.3, 0 case. Imaging changes were seen mostly in convexity, parasaggital, and falx meningiomas that were deeply embedded in the cortex. The symptom continued the until last serial observation in four cases. We used various interventions in these patients including steroid, anticoagulant, surgical removal, and hyperbaric oxygen therapy; but these therapies were not effective. CONCLUSION: LRS for each disease seems to be a safe and effective treatment. However, once serious radiation injuries occur there is no effective therapy, so it is important to make appropriate selection of patients for radiosurgery. PMID- 19999553 TI - [Multiple penetrating head injury by 8 nails: a case report]. AB - A case report of penetrating head injury due to 8 nails and a review of multiple penetrating head injury by nails were described. The patient who was a 48-year old man with a history of for psychiatric care was transferred to our emergency room on May 2007. He had shot his head with 8 nails using a nail-gun in a suicide attempt. His family called an ambulance. His conscious level was 1 on the Japan Coma Scale and 15 on the Glasgow Coma Scale with monoplegia of the right lower extremity and hypoesthesia of the right extremities. X-ray films of his head revealed 8 nails penetrating his cranium. The CT scan showed thin hematoma on the right convexity. Angiography didn't demonstrate any evidence of vascular injury. Under general anesthesia, all nails were removed after craniotomy. His post operative course was good and he was discharged home with minimal deficits. Penetrating head injury with nail-gun use is sometimes seen, but cases with multiple nails are rare. Operative strategies were discussed in the review. It also suggested the need for the care of mental or background problems. PMID- 19999554 TI - [A case of subarachnoid hemorrhage complicated by acquired hemophilia]. AB - We report a rare case of subarachnoid hemorrhage (SAH) in a man who acquired factor VIII inhibitor and suffered coagulopathy a month after disease onset. Acquired factor VIII inhibitors in patients without hemophilia is a rare disease characterized by severe bleeding as a result of antibody against factor VIII. A 61-year-old male, who had a past history of hypertension, underwent resection for colon cancer at the local hospital. Ten days after surgery he suffered septic shock that required intensive use of antibiotics. Two days after this episode, he had a sudden loss-of-consciousness attack, and was referred to our hospital with the diagnosis of SAH. Emergency angiography revealed a dissecting aneurysm at the right intracranial vertebral artery distal to the origin of the posterior inferior cerebellar artery. The aneurysm was successfully treated with endovascular parent artery occlusion. However, after a placing ventriculo peritoneal shunt a month later, he developed severe coagulopathy due to acquired factor VIII inhibitor. PMID- 19999555 TI - [A case of dural arteriovenous fistula mimicking isolated transverse-sigmoid sinus successfully treated via the transvenous route]. AB - A 66-year-old man was admitted to our hospital suffering from a left temporo occipital intracerebral hematoma with bilateral thin subdural hematomas. Neurological examinations revealed slightly decreased cognitive function. Cerebral angiography demonstrated a left transverse-sigmoid dural arteriovenous fistula (TS-DAVF) with cortical venous reflux. The main feeders of the TS-DAVF consisted of multiple branches of the left occipital artery, and the left affected transverse-sigmoid sinus was occluded at both ends, i.e. the affected sinus was isolated. Transvenous embolization (TVE) was performed under general anesthesia. A contralateral approach failed to reach the affected sinus so that an ipsilateral approach was attempted. Contrast material was injected from the left jugular bulb and the route to the affected sinus was visualized. We succeeded in passing both a microguidewire and a microcatheter through the stenosed proximal sigmoid sinus to the fistula area of the affected sinus. The affected sinus was occluded with multiple platinum coils. Post-treatment angiography revealed disappearance of the cortical venous reflux and improvement of the cerebral perfusion. It is a challenging operation to approach an affected sinus through an occluded sinus. However, some cases of isolated TS-DAVFs have the possibility of being treated by TVE via the occluded sinus similarly to cavernous sinus DAVFs. PMID- 19999556 TI - [Dural arteriovenous fistulas at the craniocervical junction fed by bilateral vertebral arteries with intracranial drainage: a case report]. AB - We report a case of dural arteriovenous fistulas (DAVFs) at the craniocervical junction, which are supplied by the radicular arteries from bilateral vertebral arteries separately, and drainaged into intracranial sinuses. A 58-year-old man with intermittent neck pain visited our hospital. T2-weighted magnetic resonance (MR) imaging showed flow voids on the dorsal surface of the medulla and upper cervical cord without any signal changes suggesting ischemia. Postcontrast MR digital subtraction angiography (MRDSA) showed early venous filling at the craniocervical junction. Angiography demonstrated bilateral fistulas near each vertebral artery penetration point of the dura matter, which were drainaged into the superior and inferior petrosal sinuses. The patient underwent suboccipital craniotomy and laminectomy of the C1, then disruption of the bilateral fistulas was performed by using micro Doppler sonography after intradural exposure of the shunt points. His symptom subsided post operatively, and MRDSA showed no abnormal vessels. Angiography performed 1 week after surgery confirmed complete obliteration of the fistulas. DAVFs at the craniocervical junction fed by bilateral vertebral arteries is extremely rare. Even in such a case, direct interruption of the fistulas using micro Doppler sonography is the most effective treatment. In addition. MRDSA could be useful for screening and perioperative studies. PMID- 19999557 TI - [A case of CNS metastasis from gastric MALT lymphoma]. AB - Recently, the incidence of primary CNS lymphoma (PCNSL) is increasing. Metastatic CNS lymphoma occurs much less than PCNSL. We report the case of a 53-year-old man who presented with CNS metastasis from gastric mucosa-associated lymphoid tissue (MALT) lymphoma. The symptoms at the time of diagnosis were dizziness and aphasia. MRI revealed a left parietal lobe tumor with a large peritumoral edema. About 4 years ago, he had suffered from gastric MALT lymphoma with a high grade component. Eradication of Helicobacter pylori led to remission of the disease 18 months after the treatment. From his past history, the brain tumor was suspected of being a metastatic lymphoma. Stereotactic biopsy revealed diffuse large B-cell lymhoma. Histopathological findings including lymphocytic subsets were almost identical between the primary gastric MALT lymphoma and metastatic brain lymphoma. Complete remission was obtained by repeated high-dose methotrexate chemotherapy. There has been no recurrence for 5 years without additional therapy. This case is probably the first report of CNS metastasis from gastric MALT lymphoma. PMID- 19999558 TI - [Unruptured saccular aneurysm at the origin of the duplicated middle cerebral artery: reports of two cases and review of the literature]. AB - We report on two rare cases of unruptured saccular aneurysm located at the origin of the duplicated middle cerebral artery. Case 1: On magnetic resonance (MR) angiography, a 56-year-old woman was diagnosed as having an unruptured right middle cerebral artery (MCA) bifurcation aneurysm. Right carotid angiography disclosed a duplicated MCA and four unruptured saccular aneurysms, including the origin of the duplicated MCA. Case 2: A 58-year-old man had a sudden onset of vertigo, and underwent MR imaging. The MR angiography detected a right internal carotid artery (ICA) aneurysm, and the subsequent angiography demonstrated duplication of the right MCA and two intracranial aneurysms; one at the origin of the posterior communicating artery (PcomA), the other at the origin of the duplicated MCA. Each aneurysm was successfully clipped through the transsylvian approach. The postoperative courses were uneventful and both patients were discharged in good condition. There have been only 19 previous reports of the duplicated MCA aneurysm in the literature. PMID- 19999559 TI - [Clinical standard of neurosurgical disorder (13) spine and spinal cord injury]. PMID- 19999560 TI - [The Schreber case and Freudian dynamic psychopathology of psychoses]. AB - We examined the diversity of diagnoses for the Schreber case, which was documented in "Memoirs of My Nervous Illness". Although Schreber became widely known as a paranoiac (or schizophrenic) patient after the publication of Freud's paper in 1911, some psychiatrists who use the operational diagnostic system have regarded him as a case of depression (or mood disorder). At present, the diagnosis of the Schreber case is divided between schizophrenia and mood disorder, which is based on the 'Kraepelinian dichotomy'. Recently, this dichotomy has been criticized as an unreasonable one, and some aspects of mood disorders are being discussed as resulting from the same biological foundation as schizophrenia. Similarly, many psychoanalysts found the same libidinal disturbance of these disorders. From the Freudian dynamic viewpoint, we conclude that schizophrenia and endogenous mood disorders have the common pathway (libidinal disturbance) in their acute phase. The findings of our literature review indicate that Freudian dynamic and economic perspective can shed a new light on the endogenous psychoses. PMID- 19999561 TI - [A case of serotonin syndrome following minimum doses of sertraline]. AB - We report a 75-year-old woman developing serotonin syndrome following minimum doses of sertraline. She showed a depressed mood, insomnia, and general fatigue and was taking sulpiride at 300 mg/day, alprazolam at 1.2 mg/day, zopiclone at 7.5 mg/day, and etizolam at 1 mg/day. As she remained symptomatic, sertraline at 25 mg/day was added. Within 14 hours of starting sertraline, the patient began to experience delirium, impaired coordination, diaphoresis, tremulousness of the upper limbs bilaterally, and agitation. Sertraline was thus discontinued, and all of the above-mentioned symptoms disappeared rapidly. Serotonin syndrome is rarely reported in patients taking sertraline in Japan. To our knowledge, ours is the second case of serotonin syndrome associated with sertraline in Japan. According to Drug in Japan, sertraline must be started at the lowest efficacious dose with slow titration and is contraindicated for patients who are taking pimozide or monoamine oxidase inhibitors (MAOIs). Also, the coadministration of sertraline with other agents such as lithium, tricyclic antidepressants, and triptans necessitates the close observation of symptoms and signs. However, our case didn't take any of these combinations, and she was administered 25 mg/day, the lowest efficacious dose. This report emphasizes that caution is needed when prescribing sertraline to elderly patients and on its coadministration. PMID- 19999563 TI - [Mental training for surgeons]. PMID- 19999562 TI - [A pioneer in occupational therapy at mental hospitals in local cities (approaches at Shichiyama Hospital at the beginning of Taisho Era)]. AB - Occupational therapy at mental hospitals in Japan began in 1901, when Shuzo Kure created two sewing rooms in the female section of Sugamo Hospital and allowed patients to sew pillowcases and hospital gowns for use at the hospital. In 1904, Sugamo Hospital added a work section, and occupational therapy became an official part of the hospital. In addition, Kure stated in The Complete Book of Japanese Internal Medicine (Nihon Naika Zensho), published in 1916, that occupational therapy had also become popular at other mental hospitals; however, he did not refer to specifics, such as information on what kind of occupational therapy was being carried out at which hospital. Shichiyama Hospital is a private facility located in Kumatori-cho, Sennan-gun, Osaka. In 1599, some 200 years before Philippe Pinel released lunatics from their chains, Sanai Honda established Soshindo at Jokenji Temple for the treatment of mental disorders using Chinese herbal remedies. Treatment using these remedies was practiced until the 1950's. Evidence suggests that Chinese herbal remedies were being used in combination with occupational therapy, such as farm work and raising poultry, from the end of the Meiji Era. Plans of the hospital in 1913 included descriptions of a farm, and entertainment and recreational areas for patients, confirming this evidence. It also supported Kure's statement about occupational therapy having become popular at mental hospitals at the beginning of the Taisho Era. In Summary, occupational therapy was practiced at mental hospitals in local cities at the beginning of the Taisho Era, and such occupational therapy in this period was significantly influenced by Shuzo Kure; therefore, I pay my respects to this great pioneer of psychiatric medicine. PMID- 19999565 TI - [Epidemiology of pleural mesothelioma in Japan]. AB - We describe the trend in pleural mesothelioma mortality rates since 1995, when the International Statistical Classification of Disease and Related Health Problems came into effect in Japan, and outline the risk factors for the disease. The number of pleural mesothelioma cases has been increasing in both genders, although male deaths predominated over female ones every year. In 2007, the latest year for which data are available, 652 deaths occurred among men and 130 among women, which was only one-fifth of that among men. The annual death rates were 0.88 and 0.20 per 10(5), respectively. The distribution of 5-year age specific deaths was skewed toward younger age, with a single peak at 70-74 years in men but at 75-79 years in women. Birth cohort analysis showed that age specific death rates from the disease became higher with younger birth year in men. Asbestos exposure causes pleural mesothelioma. Some cohort studies clarified the high mortality rate from the disease, which was reported to reach 0.5 x 10(3)/year among those with occupational asbestos exposure. Depending on the amount and variety of asbestos used in industrial plants, the range at which neighborhood asbestos exposure can increase the risk of residents developing pleural mesothelioma can be as much as 2,000 meters from an emission point. PMID- 19999566 TI - [Diagnosis and staging of pleural mesothelioma]. AB - Accurate diagnosis and staging are very important for the treatment of pleural mesothelioma. In particular, surgical indications based on diagnosis and staging must be determined carefully, since extrapleural pneumectomy (EPP) is a highly invasive surgical mode. Sufficient biopsy specimens to make a differential diagnosis of this disease should be obtained during video-assisted thoracoscopic surgery (VATS). Currently, the final diagnosis of mesothelioma is based not only on the results of hematoxylin and eosin staining but also of immunostaining using several differential markers. The International Mesothelioma Interest Group (IMIG) classification is widely used for the TNM staging of pleural mesothelioma. The IMIG classification is now applied to clinical staging with the aid of computed tomography and positron-emission tomography images, but the accuracy and usefulness of this classification for surgical indications and outcomes must be improved in future. Extended preoperative staging, for example, mediastinoscopy, contralateral VATS, and laparoscopy, are now selectively performed for the precise diagnosis of potentially resectable mesothelioma. PMID- 19999567 TI - [Diagnosis of malignant pleural mesothelioma--thoracoscopic biopsy and tumor marker]. AB - Diagnosis and treatment of malignant pleural mesotheliomav (MPM) are quite difficult. Pleural effusion or pleural thickening on plain chest x-ray is frequently observed in patients with MPM. Pleural effusion could be examined with aspiration and valuable information could be obtained. As to pleural thickening, contrast enhanced computed tomography is the most suitable modality for investigation. However, for the definitive diagnosis of MPM, video-assisted thoracoscopic surgery (VATS) is necessary. VATS biopsy offers enough tissue for pathological investigation, such as immunehistochemistry. Differential diagnosis between MPM and other malignancy is sometimes difficult, and erroneous results could be found without enough tissue. Sensitivity in the diagnosis of MPM with VATS is reported to be over 90%. Complication following VATS procedures is relatively rare. Thus VATS biopsy is strongly recommended for patients with suspected pleural effusion. Tumor markers, such as osteopontin and mesothelin, could be candidates for evaluating MPM without invasive diagnostic workup. PMID- 19999568 TI - [Therapeutic strategies for resectable malignant pleural mesothelioma]. AB - Malignant pleural mesothelioma (MPM) is a rare thoracic malignancy associated with very poor prognosis. Because no single treatment is very effective for MPM, a combination of preoperative chemotherapy, extrapleural pneumonectomy, and postoperative hemithoracic radiation is currently considered the "standard" therapy for resectable MPM. As such an aggressive protocol involves very high risk, maximum preoperative assessment for durability and curability is required. Very recently, the results of three major clinical trials of trimodality treatment for MPM have been reported from North America and Europe. The therapeutic regimens and results of these trials were similar. Although the overall results were not encouraging, surprisingly good survival rates were noted in patients who had no mediastinal node involvement and completed the entire trimodality therapy regimen. This clearly illustrates that patient selection plays a key role in MPM treatment. Two other important clinical trials are currently ongoing. The Mesothelioma and Radical Surgery (MARS) trial is a randomized clinical trial seeking to clarify the role of radical surgery in MPM treatment. An all-Japan clinical study of trimodality treatment for MPM is currently underway as a part of a national campaign entitled "Comprehensive Strategy against Asbestos-Related Diseases." PMID- 19999569 TI - [What is the role of pleuropneumonectomy in malignant pleural mesothelioma?]. AB - In this report, the results of conventional pleuropneumonectomy in malignant diffuse mesothelioma are reviewed, and the role of this aggressive surgery in multidisciplinary treatment is discussed. The surgical results may improve owing to recent advances in medical technology, although the postoperative survival time after pleuropneumonectomy has remained nearly the same for the past 40 years. The problems with pleuropneumonectomy are high mortality and morbidity rates, high local recurrence rates, and almost the same survival rates as seen after pleurectomy. The indications for this aggressive surgery are limited to stage I, II, and III disease based on an accurate staging system, with induction therapy for macroscopic radical resection and postoperative chemoradiation therapy. In addition, patients must be monitored for the development of major complications perioperatively. The natural history of this disease should be clarified to determine the effectiveness of various forms of therapy. Precise analysis of the accumulated data from many institutions will help to improve the results of pleuropneumonectomy. PMID- 19999570 TI - [Intrapleural perfusion hyperthermo-chemotherapy with cisplatin in patients with malignant pleural mesothelioma]. AB - The preferred treatment for patients with malignant pleural mesothelioma (MPM) has not been determined. In order to obtain adequate control of malignant effusion for multimodality therapy, we have introduced intrapleural perfusion hyperthermo-chemotherapy (IPHC) with cisplatin. IPHC was performed with a roller pump and heat exchanger. Cisplatin was added when the temperature stabilized to a mean of 42.5 degrees C. Dosages of cisplatin were 80 mg/m2. The circuit was filled with 2,000 ml saline. A IPHC was performed for 60 minutes under both lung ventilation. IPHC with cisplatin is feasible, easy to perform, and relatively safe. This method had brought an ideal pleural adhesion. IPHC may offer excellent local control for patients with MPM. Some literatures have reported the utility of IPHC for trimodality therapy on MPM. PMID- 19999571 TI - [Procedure performance standards of CAS in Japan]. PMID- 19999572 TI - [What we can learn from a case of medical malpractice--court rules that overtime work must be included in gynecologist's normal working hours]. PMID- 19999573 TI - [Operation for hilar cholangiocarcinoma with combined resection and reconstruction of hepatic artery]. PMID- 19999574 TI - [Issues of women surgeons in Japan]. PMID- 19999575 TI - [Various measures for problems of pancreatic surgery]. PMID- 19999576 TI - [On the regulatory role of the epsilon subunit in ATP synthase]. PMID- 19999577 TI - [Structure-based drug development and medical/biological application of cathepsin specific inhibitors]. PMID- 19999578 TI - [Structures and variable functions of TRP channels]. PMID- 19999579 TI - [Roles of signaling molecules for cell-to-cell communication during development of the central nervous system]. PMID- 19999580 TI - [Output pathways of the cyanobacterial circadian clock]. PMID- 19999581 TI - [Development of membrane-permeable peptide vectors and their internalization mechanisms]. PMID- 19999582 TI - [Roles of vesicle trafficking in Drosophila germ plasm assembly]. PMID- 19999583 TI - [DNA interstrand crosslink repair in vertebrate cells]. PMID- 19999584 TI - [Physiological functions of TRIC channels]. PMID- 19999585 TI - [Evidence-based practice guideline for the treatment of CKD]. PMID- 19999586 TI - [Longstanding intractable massive ascites caused by chronic lupus peritonitis with nephrotic syndrome: a case report]. AB - A 36-year-old woman with systemic lupus erythematosus (SLE) and nephrotic syndrome showed massive ascites. She was admitted to our hospital because of edema in both legs and a remarkably distended non-tender abdomen. On admission, massive ascites was observed in the abdominal CT scan findings. Laboratory examination of the ascites showed low levels of total protein (1.5 g/dL), albumin (0.5 g/dL) and LDH (89 IU/L), which were characterized as ascites per diapedesis. In addition, she was diagnosed with SLE and nephrotic syndrome from the clinical and laboratory findings. We treated her with steroid therapy, including methylprednisolone plus therapy. Although the serological abnormalities with SLE had normalized and urinary protein almost disappeared on the 51th hospital day, the ascites had not improved at all. These findings indicated that she had suffered from chronic lupus peritonitis, complicated with nephrotic syndrome and we had continued to treat her with prednisolone for a long time. The ascites was remarkably diminished at 220 days after admission. We believe that in addition to nephrotic syndrome, impaired vascular circulation caused by chronic lupus peritonitis might have contributed to accumulation of the massive ascites. PMID- 19999587 TI - [Case of microscopic polyangiitis presenting initially as prostatic vasculitis]. AB - A 79-year-old man was admitted to our hospital to evaluate prostatic vasculitis that was diagnosed on a prostatic biopsy performed due to an elevated prostate specific antigen (PSA)level. He complained of general malaise and had hematuria, proteinuria, and interstitial pneumonitis in the lower lung field on admission. A renal biopsy showed necrotizing crescentic glomerulonephritis. He was diagnosed as having microscopic polyangiitis (MPA) based on the presence of small-vessel vasculitis and positivity for myeloperoxidase-anti-neutrophil cytoplasmic antibody (MPO-ANCA) (465 EU). He was effectively managed with methylprednisolone pulse therapy (500 mg/day x 3 days) followed by progressive dosage tapering. In the present case, the identification of prostatic vasculitis led to the early detection of MPA, which resulted in an excellent clinical course. Prostatic involvement has been also reported in patients with Wegener's granulomatosis or Churg-Strauss syndrome. Thus, physicians should be aware of the potential for the involvement of various organs, including the prostate, in patients with ANCA associated vasculitis. PMID- 19999588 TI - [Case of anti-glomerular basement membrane antibody nephritis in a patient who was able to withdraw from dialysis and gave birth twice but underwent living renal transplantation due to progression to end-stage renal failure 15 years after onset]. AB - A 34-year-old woman with suspected rapidly progressive glomerulonephritis had been admitted to our hospital in March 1993 at the age of 19 years. Renal biopsy revealed cellular crescent formation in 24 of 26 glomeruli. Serum examination was positive for anti-glomerular basement membrane (GBM) antibody, while pulmonary hemorrhage was absent. Based on these findings, she was diagnosed with anti-GBM antibody nephritis, and treated with corticosteroid pulse therapy and double filtration plasmapheresis (DFPP) in addition to hemodialysis (HD). HD was withdrawn within 2 months. Wishing to have a baby, she had delivery in 1997 and 2000. Subsequently, her renal function gradually decreased, and she underwent an ABO-incompatible living-donor kidney transplant, with her husband as the donor, in March 2008. She has been making good progress after transplantation. Anti-GBM antibody nephritis has a poor prognosis, but renal function was maintained for 15 years in this patient, who responded well to the initial treatment. The underlying disease rarely recurs if transplantation is performed after the patient has become negative for anti-GBM antibody, anti-GBM antibody nephritis therefore seems to be a good indication for treating patients with renal transplantation. PMID- 19999589 TI - [Adult male with chronic renal failure due to reflux nephropathy that was possibly induced by neurogenic bladder since childhood]. AB - A 38-year-old male with impaired renal function and serious anemia was admitted to our hospital. He had suffered from a disorder of urination since early childhood and been diagnosed as having neurogenic bladder by an urologist when he was 20 years old. Since February 2007, general fatigue emerged and gradually worsened. In addition, he began to feel nauseous around February 2008. He visited a family doctor and was diagnosed with renal failure and anemia both of which were serious. H e was referred t o our office and admitted immediately because his blood test showed a serum creatinine level of 4.4 mg/dL and hemoglobin of 3.1 g/dL. The initial study with ultrasonograhy suggested that both kidneys contained multiple cysts for the most part, likely due to hereditary polycystic kidney disease. However, a subsequent series of diagnostic imaging tests, including computed tomography and magnetic resonance urography, determined that the cause of renal failure was most likely reflux nephropathy due to secondary vesicouretral reflux induced by the chronic neurogenic bladder. Moreover, the upper gastrointestinal endoscopic examination showed that the cause of anemia was probably the persistent bleeding from gastric antral vascular ectasia. Reflux nephropathy emerges in early childhood and slowly progresses to chronic renal failure in some cases. According to the literature, it is not rare as a cause of end-stage kidney failure even among adult populations. Diagnostic imaging of severe reflux nephropathy is apparently similar to that of polycystic kidney disease. We herein present an adult male with chronic renal failure due to reflux nephropathy, the images of which were similar to polycystic kidney disease. PMID- 19999590 TI - [Case of tuberculosis-associated hemophagocytic syndrome in a hemodialysis patient under steroid therapy]. AB - A 70-year-old woman was referred and admitted to our hospital with fever of unknown etiology. She had a past medical history of pulmonary tuberculosis. Ten weeks before admission she was diagnosed with acute renal failure caused by crescentic glomerulonephritis. Oral steroid therapy was not effective and she required dialysis. On admission, she was started on empiric antibiotic treatment, with the suspicion of bacterial infection. On the 3rd hospital day, she developed sudden hypotension and underwent direct hemoperfusion with a polymyxin B immobilized fiber. Soon after, her blood pressure normalised. Her inflammatory level apparently then improved in terms of white blood cell count and C-reactive protein, although severe fatigue and liver dysfunction persisted. On the 17th hospital day, her blood pressure went down again, accompanied by progressive pancytopenia and significant increase in serum vitamin B12, lactate dehydrogenase and uric acid. The patient was transmitted to the intensive care unit where she received bone marrow aspiration. The result revealed marked hemophagocytosis. Suspecting lymphoma-associated hemophagocytic syndrome (HPS), we administered high-dose steroid and combination chemotherapy. The treatment had no effect, and the patient died on the 21st hospital day. The autopsy demonstrated a large number of tuberculous bacilli, marked hemophagocytosis and necrosis without granuloma formation in multiple organs, leading to the pathological diagnosis of tuberculosis-associated HPS. Tuberculosis in one of the major causes for morbidity and mortality in hemodialyzed patients. It often shows atypical clinical manifestation and is difficult to diagnose. HPS in general runs a mild course unless it is lymphoma or EB virus-associated. This case seemed like bacterial infection improved with antibiotics but turned out to be a rapidly progressive tuberculosis-associated HPS. A careful examination and extensive laboratory workup is necessary to rule out tuberculosis, particularly in patients undergoing hemodialysis. PMID- 19999591 TI - [Tuberculosis among construction workers in dormitory housing in Chiba City]. AB - BACKGROUND: Tuberculosis (TB) control in a low socio-economic society is an important program for urban area of industrialized countries. Some construction workers live in Hanba, a kind of dormitory housings that have crowded living conditions, and possibly give rise to Mycobacterium tuberculosis transmission. The pulmonary tuberculosis detection rate by chest X-ray screening in Hanba is higher than the general population, and therefore TB incidence among Hanba construction worker is also estimated to be as high as that of homeless. PURPOSE: To analyze the ratio of the TB patients from Hanba in Chiba City from 1993 through 2006, and analyze the treatment outcome and speculate the factors affecting them, especially the effects of the inpatients DOTS (Directly Observed Treatment Short-course) policy introduction after 2001. METHODS: TB registration records in the Public Health Center, Chiba City, Japan, were retrospectively analyzed. RESULTS: Pulmonary TB patients from Hanba were 121 (male: 121, female: 0), representing 3.8% of the total 3179 TB patients from 1993 through 2006. Restricting to male patients aged 40-59 years-old, TB patients from Hanba were 78, representing 10.7% of 729 male TB patients of the same age groups. All of TB patients from Hanba developed pulmonary TB (PTB) and treatment outcome of chemotherapy was cured or completed: 69 (57%), defaulted or failed: 43 (36%), and died 9 (7%) respectively. When compared with PTB in Chiba and Japan, defaulted or failed was higher. In the multi-variated analysis, extensive lesions more than one lung (Adjusted odds ratio [AOR]: 0.13, 95% confidence interval [CI]: 0.04 0.37, P < 0.001) and smear-positive (AOR: 0.17, 95% CI: 0.04-0.65, P = 0.009) were negative factor for cured and completed. However, hospitalization during initial period of treatment was positive factor (AOR: 7.92, 95% CI: 1.73-36.2, P = 0.008). After inpatients DOTS introduction, the rate of cured or completed increased from 50% to 67%, and the rate of failed or defaulted decreased from 46% to 22% (P < 0.05). CONCLUSION: TB patients from Hanba, a kind of dormitory housings for construction workers, occupied 3.8% of total TB patients in Chiba City. Higher rate of defaulted or failed cases leads to poor treatment outcome in TB patients of Hanba construction workers who were possibly associated with an elevated risk of urban tuberculosis. DOTS might improve adherence to treatment and result in a decrease of failed or defaulted cases. PMID- 19999592 TI - [Efficiency of periodic QuantiFERON-TB Gold test in hospital nurses]. AB - PURPOSE: To estimate the efficiency and costs of the annual QuantiFERON-TB Gold tests (QFT) for QFT-negative female hospital nurses, and treatment of latent tuberculosis infection for QFT positive converters. METHOD: Calculation based on the models. RESULT: Risk reduction of active tuberculosis disease is estimated 48.8% on the most favorable model for the efficiency of periodic QFT tests. Total number and costs of QFT tests necessary to prevent one tuberculosis case are estimated at 4426 tests and 13.3 million yen (137 thousand US$); to prevent one bacilli-positive pulmonary tuberculosis case, 9357 tests and 28.1 million yen (289 thousand US$); to prevent one sputum-smear-positive pulmonary tuberculosis case, 22,814 tests and 68.4 million yen (706 thousand US$). CONCLUSION: Estimated cost of annual QFT tests among general female nurses in Japan to prevent tuberculosis disease is very high, and annual QFT tests could not be recommended. PMID- 19999593 TI - [Tuberculosis prevalence survey in Japan]. AB - Chest X-ray examination had been used rather soon after the discovery of X-ray by Rontgen K in 1895 as it was possible to detect chest abnormality by simple radiography. After the discovery of radiophotography independently by Abreu M and Koga Y in 1936, it was applied as a method of mass screening for TB in Japan, and Imamura A made a special lecture on "The mass screening for TB" using radiophotography in 1940 in the Annual Meeting of the Japanese Society for TB. From experiences of mass screening, it was found that there were many cases of TB who do not aware of their own disease, and to know the prevalence of TB, the screening of survey subjects by X-ray examination is indispensable. Noticing the importance of mass health examination by chest X-ray, Dr. Tanaka S, then director of information division, JATAHQ, edited a book entitled "How to carry out mass health examination for TB" in 1951, then he moved to the Ministry of Health and Welfare and engaged in the preparation of the first TB prevalence survey. Random sampling technique was already developed, and health center network covering the whole country was already completed in early 1950s. Using these background, the first TB prevalence survey was conducted in 1953. TB Prevalence Survey Committee was organized asking cooperation of experts in TB, epidemiology and statistics, and the survey in sampled area was carried out by a survey team headed by the director of health center in charge of the sampled area. The survey teams engaged in the survey with enthusiasm, and the rate of response to the survey was 99.3%. The result of this survey was published in the WHO Bulletin, 1955. After the survey in 1953, the following prevalence surveys were carried out in 1958, 1963, 1968 and 1973. Outline of these surveys was shown in Table 1, and the rate of examination was high in all, except the survey in 1973. In this year, TB prevalence survey was carried out in conjunction with the national nutritional survey and the national mental health survey, and unfortunately, there were some opponent groups against the national mental health survey, which affected the rate of response to the TB survey, too. In addition to the 5 prevalence surveys, one thirds of the survey population in 1953, 1958 and 1963 was surveyed in the next year to know the incidence of TB. Follow-up survey on active TB cases found in the 1953 and 1958 survey was carried out in 1964, and similar follow-up survey was carried out in 1968 for active TB cases found in 1953, 1958 and 1963 surveys. Moreover, survey subjects excluding active TB cases in 1968 were followed up until 1973, and the incidence of newly registered TB cases during this period was surveyed. Summarized results of TB prevalence surveys are reported. The first survey was carried out in 1953, and had been repeated every 5 years until 1973. As national TB control program (NTP) under new TB Control Law had been implemented since 1951, the results of 5 surveys clearly indicated the outcome of NTP of Japan. Age-specific prevalence of active TB in 5 surveys is shown in Fig. 1. Due to advances in chemotherapy, there was certain difference in the definition of active TB in 1953 and 1958, and chemotherapy was indicated more widely for those with TB pathology in lung in 1958. Comparing the age-specific prevalence of active TB in 1953 and 1958, the prevalence decreased in 1958 below 35 years of age, and increased above 35 years. The decline in the prevalence of active TB in age groups below 35 in spite of widening of definition of active TB in 1958 indicated the efficacy of TB control with mass screening and BCG vaccination and treatment for detected cases. As the definition of active TB had been unchanged since 1958 up to 1973, the decline in the prevalence of active TB seen in all age groups clearly indicated the achievements of NTP. Overall trend of prevalence of active TB, cavitary TB, bacillary TB and smear+PTB in 5 surveys is shown in Fig. 2 together with epidemiological figures obtained from vital registration, namely incidence of TB, prevalence of active TB at the end of the year and TB mortality. In analyzing the results, we have to take note of the difference in bacteriological examination methods. As laryngeal swab method was used in 1963 and 1968, culture positive rate was lower and no information about smear examination, however, from 1958 to 1973, all indices had declined exponentially with similar speed including prevalence of smear +PTB and bacillary TB if results in 1958 and 1973 were connected directly. Based on this results and the fact that marked decline in the prevalence of active TB requiring much larger sample size for the survey, TB prevalence survey was stopped, and data from vital register has been used since then to evaluate the TB situation. Results of 3 incidence surveys were shown in Fig. 3. Shift of higher incidence from younger age groups to higher age groups was clearly shown from 1954 survey to 1964 survey. The results of routine follow-up by vital registration of 1968 survey population excluding TB cases found in 1968 survey were shown in Table 2, and high risk groups were clearly shown in this table. As routine X-ray examination was done by radiography in 1963 survey, it was possible to pick up any slight TB pathology, and the age-specific prevalence of any TB finding, that of healed findings and of calcified lesions were shown together with BCG vaccination coverage in Fig. 4. In the age groups above 40, the prevalence of any TB finding, as well as of healed and calcified findings was very high, while the coverage of BCG vaccination was below 20%. BCG vaccination was started in Japan in 1943, and those above 40 years of age in 1963 were then already 20 years of age or above, and only few were vaccinated with BCG when BCG vaccination was expanded to cover higher age groups thereafter. TB prevalence survey has now come to be used as one of methods to estimate the incidence of TB under the impact of HIV/AIDS epidemic, and its significance is now re-evaluated. PMID- 19999594 TI - Tuberculosis: a new vision for the 21st century. AB - Tuberculosis is a global problem that we can't afford to keep ignoring. In 2006, tuberculosis killed 1.7 million people--almost twice as many people as malaria- and it is the leading cause of death among people living with HIV/AIDS. This is all the more tragic because these deaths are preventable. For a long time the world thought that we had defeated tuberculosis, but just because tuberculosis doesn't make headlines doesn't mean it has gone away. The fact is that tuberculosis is getting worse, as complacency and lack of adequate tools and funding fuel the disease and the spread of drug resistance. Drug resistant tuberculosis is the wake-up call, it is an airborne epidemic of increasingly untreatable disease. Drug resistant tuberculosis develops when tuberculosis patients take low-quality drugs, do not finish their full course of treatment, or pass drug resistant tuberculosis from one person to another. In 2007, there were approximately 500,000 cases of drug resistant tuberculosis globally. MDR-TB is resistant to the two most commonly used first-line TB drugs, and requires long, complex and expensive treatment. XDR-TB is resistant to first- and second-line drugs, severely limiting treatment options. While progress is being made, much more is needed. Basic tuberculosis control is one of the most cost-effective interventions in global health. Appropriate treatment can save a life and stop the spread of disease for US$14. It is essential that countries implement the World Health Organization's (WHO) internationally recommended Stop TB strategy, which includes DOTS. But due to outdated tools and methods, DOTS alone is not enough. The remarkable fact is that global control of tuberculosis, a disease that kills someone every 20 seconds, depends upon a 125-year-old test, an 85-year old vaccine and drugs that take six months to cure and haven't changed in four decades. To successfully treat tuberculosis and prevent resistance, we need to use current tools better and accelerate the development of new tools for the future. Simple improvements in tuberculosis control, such as expanding the use of under-utilized technologies, can have enormous impact. Fixed-dose combinations have existed for over 25 years, and could help ensure that more patients complete treatment; yet globally, only 15 percent of patients are using them. We also need new drugs, vaccines and diagnostics, as well as innovations in tuberculosis control and case management. Better diagnostics are already available, and new drugs and vaccines are coming. But more commitment and resources are needed. Better prevention and control of tuberculosis is the surest way to stop drug resistance. To ensure that drug resistance does not pose a wider threat, we need to employ a number of equally important approaches. These include improved basic tuberculosis control, increased use of underutilized technologies such as fixed dose combinations, and new technologies and health systems innovations. At the same time, we should expand access to M/XDR-TB treatment and diagnostics for those who already have drug resistant tuberculosis. Some of the most innovative solutions can come from the private sector and through partnerships. An untapped market of two billion people carries the tuberculosis bacterium. Since tuberculosis requires a comprehensive approach, companies should also explore opportunities to work together and pool complementary technologies to ensure new tools are used most effectively. Japan is poised to play a leading role in the discovery, development and delivery of tuberculosis solutions in the 21st century. PMID- 19999595 TI - [Fight against urban tuberculosis problems and program effects in Osaka City]. AB - In Osaka City, tuberculosis (TB) incidence rate (909.7) was already twice as high as national average in 1950. It had been reduced thereafter but was stagnated above 100 in the 1980's and 90's and it has become the worst in the country. To improve the situation, the 10 year plan of TB control was formulated and practiced since 2001. The plan mainly consists of proper treatment promotion to use 4 drugs including pyrazinamide at initial phase, introduction of DOTS, and expansion of chest X-ray screening among homeless which leads to early case finding. As an effect of plan after 8 years have passed, implementation rate of DOTS virtually reached 80% as targeted. As a result, defaulter and failure rate was reduced from 13.7% to less than 5%. The rate of retreatment cases among homeless patients was reduced from 43% to 17%. Multidrug resistant rate among patients without previous treatment was reduced from 1.1% in 2001-2004 to 0.6% in 2005-2008. And overall TB incidence rate became half (50.6) in 2008 from 100 per 100,000 population before the plan started. Factors contributing to the above success seem to be as follows: formulation of long term control plan with indicators, integration of tuberculosis diagnostic committee, introduction of DOTS, strengthening and expansion of chest X-ray screening for the homeless, and unified implementation of contact examination in congregate settings, etc. Moreover patient management has been discussed in the regular meeting on cohort analysis at all public health offices in the city. When they had any questions on the treatment, they can now communicate with medical staff as necessary. It was indispensable to collaborate with welfare department and coordinate with NPOs to solve problems regarding homeless patients. PMID- 19999596 TI - [Medical economics in tuberculosis management: what will come after the TB ward?]. AB - Current unprofitability of medical services in tuberculosis (TB) ward in Japan has been induced by low medical fee and long-term hospital stays, aggravated by unoccupied beds due to the decrease in the number of patients. For the solution of this issue, the increase of medical fee, shortening of the length of hospital stay and drastic reduction of oversupplied beds are essential. An increment of medical fee by the change in the system would be appreciated, but even under the current system, the balance between revenue and expenditure could be obtained by reducing the length of hospital stay toward 4 weeks, and the elimination of deficit in TB ward could be accomplished by these efforts; shortening of length of hospital stay and reduction of TB beds. Although the latter might result in the difficulty of sustaining TB wards, these patients could be treated in the infectious disease ward. The integration of TB Control Law to Infectious Disease Control Law suggests that TB is not a special disease in Japan. If a true "short course therapy" era would be realized by novel anti-tuberculosis drugs, a dramatic change in TB management would occur in the near future. PMID- 19999597 TI - [Tuberculosis annual report 2008--series 2. TB in foreigners]. AB - Statistics on tuberculosis (TB) in foreigners have been obtained since 1998 in Japan. The number of foreign TB patients increased from 739 in 1998 to 945 in 2008. In contrast, the number of Japanese TB patients decreased during this period and hence the proportion of foreign TB patients increased from 2.1% in 1998 to 3.9% in 2008, excluding those of unknown nationality. Especially, the proportion of those aged 20-29 years increased greatly from 9.1% in 1998 to 26.3% in 2008. Although the number of nationalities was 47, the majority of patients were from China (27.7%), the Philippines (24.8%) and Korea (10.2%) in 2008. The number of foreign TB patients aged 20-29 years was 468, accounting for 49.5% of all foreign TB patients in 2008. Seventy-seven percent of foreign patients aged 20-29 years had developed TB within 5 years of entering Japan. The equivalent proportion was 49% of those aged 30-39 years and 32% of those aged 40-49 years. Regarding occupation, 39.7% of foreign patients aged 20-29 years were full-time workers, 28.6% were students and 13.7% were part-time workers. PMID- 19999598 TI - [Complications during anaesthesia for diagnostic and interventional cardiac procedures in children with congenital heart defects]. AB - BACKGROUND: Cardiac catheterisation, while enabling dynamic evaluation of the cardiovascular system, is also commonly performed for interventional procedures in all age groups. The aim of this study was to analyse the incidence and spectrum of complications occurring during general anaesthesia in children undergoing cardiac catheterisation. METHODS: We retrospectively reviewed the medical charts of 1622 consecutive patients, anaesthetised for 817 diagnostic, and 805 interventional cardiac procedures. Data on patient- and procedure-related variables affecting the risk of complications was collected. We analysed the influence of age, physical status, type of heart defect, type of procedure, and anaesthetic protocol on the incidence of complications. RESULTS: Serious complications occurred in 5 patients undergoing diagnostic procedures and in 24 undergoing interventional procedures, with infants being the most affected (21 cases). The most frequent complication was acute heart failure related to the catheterisation. In 2 cases, pneumothorax followed central venous catheter insertion. There were 14 cardiac arrests requiring cardiopulmonary resuscitation, resulting in three deaths. CONCLUSIONS: The results show that diagnostic cardiac catheterisation and interventional procedures are associated with a low risk of complications (1.78%) and a low mortality rate (0.18%). PMID- 19999599 TI - [Effects of sevoflurane versus target-controlled infusion of propofol on haemodynamics during elective breast surgery in healthy women]. AB - BACKGROUND: Haemodynamic changes during general anaesthesia are still being widely investigated. Although propofol and sevoflurane are commonly used anaesthetic agents, there is a lack of comparative studies assessing their influence on haemodynamic parameters. We have assessed whether the effects of target-controlled propofol anaesthesia on HR, MAP and cardiac output, are different from those seen with a sevoflurane-based protocol. METHODS: ASA I and II females, scheduled for elective breast surgery, were studied. After premedication with midazolam and intravenous induction with propofol, fentanyl and vecuronium bromide, general anaesthesia was maintained with either 1 MAC sevoflurane in oxygen/air (group S), or a target-controlled infusion of 3 micromL(-1) propofol (group P). Both groups received additional fentanyl. Heart rates and mean arterial pressures were noted and compared. Stroke volumes, cardiac outputs and cardiac indexes were measured every 3 min, using a non invasive carbon dioxide rebreathing method (NICO). RESULTS: Fifty-seven patients were enrolled in the study. Comparisons between the groups revealed a significant decrease in mean heart rate during maintenance of anaesthesia with propofol (p<0.05), but not with sevoflurane. Although cardiac output and cardiac index were lower in group P towards the end of analysis, no statistically significant differences in HR, MAP, SV, CO or CI were found between the groups. CONCLUSION: The haemodynamic effect of a target 3 micromL(-1) propofol infusion did not differ significantly from that observed with 1 MAC sevoflurane. PMID- 19999600 TI - [Neuromuscular block monitoring for optimisation of conditions for endotracheal intubation]. AB - BACKGROUND: The choice of an appropriate moment for endotracheal intubation is essential to avoid serious motor and cardiovascular reactions during laryngoscopy and tube insertion.The purpose of the study was to compare the effects of intubation on laryngoscopy conditions and cardiovascular response, when choice of the moment for intubation was directed by either clinical or train-of-four assessment. METHODS: Adult ASA I patients, scheduled for lumbar disc hernia surgery, who received 0.15 mg kg(-1) of cis-atracurium for muscle relaxation, were divided in two groups. Patients in group I were intubated when the attending anaesthesiologist assessed muscle relaxation to be adequate. Patients in group II were intubated when there was no visual response to train-of-four stimulation of the ulnar nerve. RESULTS: Forty-five patients were enrolled in the study. The mean time for intubation was 162.3+/-35 sec in group I and 339.3+/-73.7 sec in group II. Adequate and excellent conditions for intubation were achieved in all patients of group II, compared to only 53% of patients in group I. Heart rate and arterial blood pressure immediately after intubation were significantly lower (p<0.001) in group II. CONCLUSION: The objective assessment of neuromuscular relaxation priorto endotracheal intubation provides better conditions and minimization of cardiovascular reaction. PMID- 19999601 TI - [A comparison of the optical AirTraq and the standard Macintosh laryngoscope for endotracheal intubation in obese patients]. AB - BACKGROUND: Endotracheal intubation in morbidly obese patients is usually difficult and may lead to traumatic complications. METHODS: We compared the median time needed for endotracheal intubation, between a newly introduced device, the AirTraq optical laryngoscope, and a standard device, the Macintosh blade. The study group comprised adult patients scheduled for elective abdominal surgery, with a BMI >39. RESULTS: Sixty-eight patients were enrolled in the study. The time to successful insertion of an endotracheal tube was 29 and 49 sec in the AirTraq and the Macintosh groups, respectively. In the AirTraq group, additional manoeuvres were required to improve the laryngeal view in 7 cases. In the Macintosh group, an elastic guidewire was used to facilitate endotracheal intubation. No traumatic complications were observed in either group. CONCLUSION: The optical AirTraq laryngoscope proved to be more useful than the Macintosh blade for faster and easier intubation in morbidly obese patients. PMID- 19999602 TI - [Open versus percutaneous repair of abdominal aortic aneurysm]. AB - BACKGROUND: The mortality in aortic abdominal aneurysm surgery remains in the 2 7% range. The aim of the study was to compare the results and safety of surgical aneurysm resection with grafting under general anaesthesia, using percutaneous insertion of a stent under local anaesthesia and intravenous analgesic sedation. METHODS: The data from patients scheduled for elective subrenal abdominal aortic treatment was analysed retrospectively. Patients were allocated to two groups, according to their general condition, ASA status, presence of other pathologies and anatomical conditions. Patients in group I underwent open surgery with general anaesthesia; patients in group II received stents implanted using local anaesthesia and intravenous analgesic sedation. RESULTS: One houndred twenty seven patients were allocated either to group I (n=87) or to group II (n=40). Blood pressure, heart rate, the need for catecholamines, blood loss and fluid replacement were all higher in group I. More than 50% of patients from this group required mechanical ventilation in the early postoperative period, as well as longer hospitalization. All patients in group II survived, while 6 patients from group I died from postoperative complications. CONCLUSIONS: Endovascular repair of aortic abdominal aneurysms under local anaesthesia combined with i.v. sedation is a safe alternative in high-risk patients. PMID- 19999603 TI - [A survey of basic resuscitation knowledge among medical personnel of a paediatric hospital]. AB - BACKGROUND: The European Resuscitation Council (ERC) algorithms for cardiopulmonary resuscitation have undergone many changes, the latest version being published in 2005.To establish the level of general knowledge of resuscitation algorithms, we audited physicians and nurses in a tertiary paediatric hospital. METHODS: The audit was performed among 64 physicians and 54 nurses. They were allocated to the following subgroups: surgeons, paediatricians, surgical nurses, and non-surgical nurses. A written open examination was conducted, containing response questions on basic resuscitation of adults, children and infants. The participants also completed a questionnaire concerning their knowledge of current guidelines and their activity in continuing education. RESULTS: The vast majority of those audited were not familiar with the 2005 ERC guidelines. Most of the physicians and nurses were not able to differentiate between an adult and a child.They did not know the recommended number of artificial breaths to be delivered to a patient with preserved cardiac function, or the number of resuscitation cycles (delivered by one or two rescuers). They could not describe the open airway manoeuvres and volumes of artificial breaths. Depending on subgroups, only 14.3-84.6% of participants knew how to deliver chest compressions to adults, 21.4-80.8% knew how to deliver them to children, and 19.2 75.4% knew how to deliver them to infants. 17.8-23.1% of nurses and 28.6-47.4% of physicians had completed basic life support courses. 0-14.3% of nurses and few physicians declared a familiarity with the 2005 guidelines, but only 35% of them were aware of the latest modifications. CONCLUSION: The medical staff of a paediatric hospital was not adequately trained in cardiopulmonary resuscitation and their knowledge about current recommendations was minimal. The authors suggest that life support courses should be mandatory for all physicians and nurses. PMID- 19999604 TI - [Methylene blue therapy for vasoplegia after cardiac surgery]. AB - BACKGROUND: Systemic inflammatory response syndrome (SIRS) may be associated with critically decreased vascular resistance (vasoplegia syndrome) and become resistant to noradrenaline. Such a condition may be successfully treated with intravenous methylene blue. CASE REPORT: A 73-year-old patient with acute myocardial infarction and cardiogenic shock was admitted to our centre after failed percutaneous coronary interventions to the left anterior descending coronary artery and underwent emergency coronary revascularisation. After surgery, his ejection fraction was 25%. X-ray revealed pulmonary oedema. The patient became severely hypotensive with marked vasoplegia and metabolic acidosis, despite incremental doses of adrenaline and noradrenaline. 2 mg kg(-1) of methylene blue was administered, which quickly stabilized the patient's general condition. CONCLUSION: Methylene blue applied to vasoplegic patients can effectively increase systemic vascular resistance and restore satisfactory organ perfusion. PMID- 19999605 TI - [Accidental intravenous injection of formaldehyde]. AB - BACKGROUND: Formaldehyde can be found in operating theatres where it is used for preservation of biopsied tissues. Several misuse accidents have been described previously.We present a case where formaldehyde was mistakenly injected intravenously. CASE REPORT: A 33-year-old man, scheduled for excision of a knee meniscus under spinal anaesthesia, was to receive an intravenous antibiotic at the end of surgery. The attending anaesthesiologist received a vial of cephazolin, marked with the patient name from a scrub nurse and injected its contents intravenously. Immediately after injection, the patient complained about strong pain at the site of injection and started to cough.The vial was checked again and a piece of meniscus preserved with 4% formaldehyde was found inside. It was intended to be offered to the patient on departure. The possible amount offormaldehyde injected was 400 mg (a lethal dose has been described as 12 g). The patient, despite the lack of cardiorespiratory failure, was intubated, ventilated and dialysed for six hours, and then extubated without further consequences. His biochemical markers remained in the normal range. Based on the case as described, the possible medico-legal consequences of poor organisation and preventive measures are discussed. PMID- 19999606 TI - [Transversus abdominis plane block]. AB - A substantial component of the pain experienced by patients after abdominal surgery is derived from the abdominal wall incision.The transversus abdominis plane (TAP) block is a novel approach for blocking the abdominal wall neural afferents via the bilateral lumbar triangles of Petit.The block has been described by McDonnell and colleagues, and can be used for intra and postoperative analgesia in patients having surgery in the lower abdominal region, especially for caesarean section, inguinal hernia repair and laparoscopy. The technique of blind and ultrasound-controlled blockade is described and discussed in detail. PMID- 19999607 TI - [Safety of metamizole and paracetamol for acute pain treatment]. AB - Postoperative analgesia should be adjusted to current needs of a patient. Non opioid agents are recommended, wherever possible: both non-steroidal anti inflammatory drugs (NSAIDs), metamizole or paracetamol may be useful for treatment of acute pain. The use of metamizole is associated with such complications as bone marrow damage (agranulocytosis, aplastic anaemia), chronic interstitial nephritis, gastro-intestinal disturbances, etc. Paracetamol, currently also intravenous, is likely to cause hepatocyte damage, renal necrosis, as well as vomiting, diarrhoea and skin reactions. Opinions about metamizole are controversial. In some countries, metamizole was withdrawn already in the 70ties of the previous century; in others, it is still widely used. According to the Uppsala Monitoring Centre, WHO Collaborating Centre for International Drug Monitoring, the number of adverse effects registered in the years 1978-2009 (March) was 14441 for metamizol and 67581 for paracetamol. Modern multimodal analgesia should be based on a good combination of analgesics. Both metamizole and paracetamol may be used for such a purpose, yet in the lowest effective doses, within the shortest needed time and once evident contraindications have been considered. Safety of both drugs is several times higher than that of commonly used NSAIDs. PMID- 19999608 TI - [Difficult airways--why do we need algorithms]. AB - The difficult airway and/or intubation are still one of the most important challenges for anaesthesiologists. Recent clinical trials reported a decrease in the number of fatal cases associated with a difficult airway; however, these complications are still among the major causes of severe morbidity and mortality in anaesthesiology. Severe anaesthesiological complications including serious hypoxia, brain damage and death occur mostly during induction of anaesthesia, and are often related to a difficult airway. The term is often misinterpreted, and relates to failed intubation, difficult tracheal intubation, difficult laryngoscopy, and/or difficult mask ventilation. Difficult intubation is not easy to anticipate. The majority of prognostic methods, i.e. assessment scales accepted in clinical practice, are not satissfactory. Up to 40% of tracheal intubations preoperatively anticipated as difficult are performed without problems. In some cases, however, the procedure proves difficult only on direct laryngoscopy. This review presents current reports and statistical analysis of serious airway-related incidents in medical practice, and indicates the need for the introduction of mandatory guidelines for difficult airway assessment and management in clinical practice. PMID- 19999609 TI - [Algorithms for unanticipated difficult endotracheal intubations in adults]. PMID- 19999610 TI - [Internal jugular vein cannulation]. PMID- 19999611 TI - [Primary reconstructive operation in surgical treatment of perinatal brachial plexus palsy in children]. AB - Perinatal brachial plexus palsy (PBPP) constitutes a serious medical problem. PBPP treatment consists in conservative and operative procedures. The latter one is usually divided into primary operative procedures, conducted in the initial stage of child's life (aiming at suture of brachial plexus nerves in the early stage of life) and secondary (aiming at elimination of functional limitations, osseous deformations, articular contractures occuring at later stages of life). A significant but also controversial aspect arises at determining the date of operation performance and also at defining factors classifying a child for surgical operation and details concerning the application of definite operative method. The following microsurgical methods are applied in microsurgical reconstruction of PBPP: neurolysis (external and internal), end-to-end nerve suture, free nerve grafts. The most complicated are injuries which result in root avulsion from the spinal medulla. In such cases the avulsed roots are connected to nerves of cervical plexus, applying the method of neurotization. The reason of numerous controversies around the choice of treatment method for PBPP patients is the desire to balance procedures aiming at the proper functioning of the patient in future. PMID- 19999612 TI - [Use of questionnaires in outcome measurement in hand surgery]. AB - An assessment of outcomes of the treatment of the diseases and injuries to the hand should include both objective parameters, i.e. range of motion or grip strength, and subjective parameters i.e. intensity of complaints or dexterity of the hand in various tasks of daily activity. Questionnaires were developed to evaluate subjective parameters in objective and systematic manner. They indeed allow us to better apprehend what is important from our patients' perspective. Questionnaire (called also form or test) is a list of queries concerning the issue being a subject of the assessment. The queries are designed in the way allowing answers to imply an important and possible objective information on the problem of interest. The objective of this article was to attract notice on the problems associated with the use of questionnaires in a research, with particular attention on hand surgery. Authors referred to the parameters which characterise these instruments: responsiveness, validity and reliability. Brief descriptions of most commonly used forms were provided, including DASH, Levine, PEM, Michigan and CISS. Shortcomings and pitfalls associated with the use of questionnaires and possible sources of bias were emphasised, e.g. difficulties in precise assignation of scores, answering some particular questions and accuracy of the completion of the form. Attention was paid on the choice of the questionnaire to be well-matched for an aim of a given research project both in substantial and formal manner. An opinion was supported that modem research projects measuring outcomes in hand surgery must include questionnaires as an obligatory component of a complex assessment. PMID- 19999613 TI - [Total hip arthroplasty in rheumatoid arthritis: results of 12-years follow-up]. AB - The aims of retrospective cohort study were: analysis of general and local factors, evaluation of clinical and radiographic results and presentation of authors' experience in total hip arthroplasty in rheumatoid arthritis. Study group consists of 222 consecutive total hip arthroplasties (205 cemented and 17 cementless) made in 216 patients. Mean patients' age was 58.4 y.o (range: 34-77 y.o.). In study group were 192 women and 24 men. Mean preoperative function was poor: 28.4 points (range: 24-32 p.). Clinical evaluation was made with Harris hip score, and radiographic one with criteria of Joined Committee of Hip Society, AAOS and SICOT. Clinical results of the study group were as follow: excellent--53 (23.9%), good--128 (57.6%), fair--34 (15.3%) and poor--7 (3.2%). Mean postoperative function was good: 86.8 points (range: 28-94 p.). There were following radiographic results of study group: good--176 (79.3%), fair--28 (12.6%) and poor--18 (8.1%). Complications another than aseptic loosening were observed in 56 (25.2%) hips. CONCLUSIONS: 1) general and local factors make total hip arthroplasty in rheumatoid arthritis more difficult procedure than the same one in osteoarthritis, 2) log-term survival rate of hip replacement in rheumatoid arthritis mainly depends on proper bone stock reconstruction, 3) 80-85% of excellent and good clinical and radiographic results are achieved at 12-year follow-up, 4) incidence of local and systemic postoperative complications are similar to osteoarthritis population, 5) preoperative autologous blood donation does not decrease demand of postoperative blood transfusion. PMID- 19999614 TI - [Realloplasty of the acetabular component of the hip joint endoprosthesis without mechanical support systems]. AB - This work presents the tactics of surgical procedures in 40 patients who underwent revision of the hip joint due to isolated loosening of the acetabular component of the endoprosthesis. The studied group were operated between 2004 to 2006 without the use of mechanical support systems. Realloplasty of the acetabular component involved the evaluation of: etiology of acetabular loosening, concomitant diseases in the operated patients, and the grade of acetabular defects according to Paprosky's classification. In this work we analyze the type of both the loosened acetabulum and that used in revision, and discuss complications. Finally, we present a clinical evaluation of the operated patients according to Harris scale before revision surgery and six and twelve months after, with good early results. PMID- 19999615 TI - [Primary total knee arthroplasty with structural autologous bone grafting of medial tibial condyle defect in elderly patients: a preliminary report]. AB - AIM OF THE STUDY: The aim of the study is to assess the early results of primary total knee arthroplasty with reconstruction of medial tibial condyle with autogenous bone grafting in elderly patients. MATERIALS AND METHODS: The study was conducted on patients treated with primary total knee arthroplasty. Eight subjects (eight knees) aged 63-82 (mean, 73) with mean varus alignment 18 degress (range, 10-30 degrees) required restoration of medial tibial condyle. Autogenous bone grafting from resected joint ends was used and stabilized with cancellous screws. Patients were surveyed pre- and postoperatively with questionnaires, such as: Knee Injury and Osteoarthritis Outcome Study (KOOS) and SF-36 Health Survey, version 2. Incorporation of the bone graft were evaluated in radiographs. Subsequent folow-up assessments were performed 3, 6 and 12 months after surgery. RESULTS: Postoperatively, mean range of motion in the knee joint improved from 99 to 124 degrees and pain relief was observed. The largest improvement in KOOS and SF-36 scores was seen for activities of daily living, pain, quality of life and physical functioning, bodily pain domains, respectively. The analysis of radiographs showed incorporation of the bone grafts with no lysis. There was no evidence of graft collapse or tibial component loosening. CONCLUSIONS: Autogenous bone grafting is valuable material to fill defects in the proximal tibial plateau. Volume of resected grafts and incorporation time are limitations for this technique. On the basis of our observations, we presume that this method is effective in elderly patients. PMID- 19999616 TI - [Subtalar arthrodesis through single medial approach in the treatment of acquired, fixed flatfoot deformity--preliminary report]. AB - Authors presented the results of subtalar and in some cases talonavicular arthrodesis through single medial approach in the treatment of fixed planovalgus foot deformity. The procedure was performed in 15 feet with fixed hindfoot valgus deformity. In all cases, after achieving correct position, internal fixation of the subtalar and talonavicular joints was done with single, canulated, compressive screw 6.5 and 4.5 mm (Unima, Eos) respectively. Solid fusion was achieved within 8 to 12 weeks after surgery. Arthrodesis of the subtalar and talonavicular joints through a medial approach is usefull procedure in achieving correction and fusion of these joints while avoiding soft tissues healing problems often encountered with traditional lateral approaches. PMID- 19999617 TI - [Pedobarographic evaluation of the foot after Keller's procedure for Hallux valgus and Hallux rigidus]. AB - Hallux valgus and hallux rigidus are common foot deformities. One of possible procedures is Keller's surgery. The purpose of this study is to evaluate the use of pedobarography for postoperative evaluation of the foot. Group. We examined 54 patients aged 48 to 82 (average 60.4) after surgical treatment of Hallux valgus (78 feet) or Hallux Rigidus (15 feet). Methods. Retrospective pedobarographic evaluation 1 to 11 years after surgery was performed using PEL-38 system. Clinical results. All patients had markedly increased pressure under the 2nd and 3rd metatarsal heads comparing to other forefoot regions. Only 17% of feet had distal dynamic transverse arch during stance phase of gait. There was severe impairment of function of the great toe in entire group. In 82.7% of feet there was no ground contact of the great toe in dynamic evaluation of the stance phase. Conclusion. Pedobarographic examination has good value in postoperative functional assessment of the foot. It's a valuable addition to physical and radiological examination. PMID- 19999618 TI - [Pain intensity in patients treated by the external fixation]. AB - One of the inherent features of the external fixation is pain occurring during the whole treatment process. The aim of this paper is evaluation of the level of pain intensity during particular stages of treatment and defining the correlation between pain intensity and type and also localization of fixator, etiology, type of treatment and patient's life activity. We analyzed 64 patients treated by external fixation of the lower limb in the age between 13 to 72 (mean 23.7). The pain intensity was evaluated four times by the NRS protocol (Numeric Rating Scale) in active and passive situation. It is stated that there is a different pain pattern in time depending on the type of treatment and personal life activity. Lengthened and patients in full time education are characterized by rapid increase in pain level in the first period of treatment. Patients with stabilization of non-union or fracture and professionally inactive had lower fluctuations of pain intensity. Localization of apparatus has influence on pain intensity level- greater pain was noted in the tibial group. Etiology influences on pain intensity. Increase of pain intensity in patients with non-union is lower in comparison to patients treated for the other indications. PMID- 19999619 TI - [Analysis of kinematic parameters of normal-paced gait in subject after posterior cruciate retention and substitution total knee replacement--leading article]. AB - The main goal of this study was to estimate the variability of gait parameters (angular changes) in patients after cruciate-retaining and cruciate-substituting total knee arthroplasties. Angular changes of the knee joint in three planes were taken into consideration. The three-dimensional analysis of locomotion pattern was carried out on 18 patients aged 60-74, using the Vicon system. 8 patients were after posterior cruciate-retaining (CR) total knee arthroplasty and 10 after posterior-stabilized (PS) arthroplasty. The control group consisted of 30 healthy subjects 50-70 years old. The results showed, that CR patients walked on flexed knees and the varus alignment in stance phase was observed. Increased internal rotation was also visible. The movement analysis of PS knee joints was similar to CR group but the angular changes in frontal and transversal planes were more increased compared to the norm. PMID- 19999620 TI - [Arthroplasty of both hips and knees]. AB - INTRODUCTION: The total joint replacement of the four great human articulations concerns patients with extreme functional impairment of the locomotor system (musculoskeletal system). Most frequently this regards patients with rheumatoid arthritis (RA) or the ones treated operatively in childhood for DDH (Developmental Dysplasia/Dislocation of the Hip) or patients with idiopathic arthritis of the hip and knee. MATERIALS AND METHODS: Eight patients underwent total join replacement of both hips and both knees (four-joint arthroplasty) between 1986 and 2006. For six patients the cause of treatment was RA, for one patient it was an implication of DDH and for the last one it was an implication of idiopathic osteoarthritis of hips and knees. The operative treatment was a multistage process (during one operation only one joint was replaced). The mean age of the patient at the time of the last procedure was 51 years (range 36 to 70 years). The mean follow-up, of four-joint replacement, was 6 years). The mean follow-up of four-joint replacement, was 6 years. We based our outcome evaluation on clinical (HHS, KSS, WOMAC) and radiological examination. RESULTS: The average preoperative Harris score for the group of patients was 35.3, WOMAC score--79.5, and Knee Society Score--38.1. After an average of 6 years follow-up all hips and knees were considered excellent, with average Harris score of 99.4, WOMAC Score of 4.5 and Knee Society Score of 98.3. All patients had increased function and decreased pain. The radiograms of all patients revealed that the acetabular and femoral components and the femoral and tibial ones were correctly positioned with no radiographic evidence of loosening in the last examination. CONCLUSION: Clinical and radiological evaluation of our material showed that total replacement of both hips and knees allows regaining good lower limb function, which helps the patients staying less dependant on the surrounding environment. PMID- 19999621 TI - [Pain mechanisms in Complex Regional Pain Syndrome Type 1--a review]. AB - The phenomenon of pain in the course of Complex Regional Pain Syndrome Type 1 is interesting, because its mechanism of its development and maintaining has not been definitively explain. The objective of this article was to review of contemporary knowledge in this issue. The following hypotheses were mentioned: 1) sensitization of dorsal root neurons on afferent barrage form site of injury, of which each, also non-nociceptive, can induce an activation of the nociceptive pathway; 2) interactions between the nervous and immune systems, when pain stimulation as a result of injury or inflammation activate microglia cells in central nerve system, what probably is responsible for development of spontaneous pain; 3) the role of of glutamatergic N-metyl-D-asparagin receptor (NMDA), which increased activity reduces pain experience due to changes of conductance and number of AMPA receptors, that mediate fast pain transmission; 4) the phenomenon of allodynia, when a touch stimuli, temperature or limb position change is experienced as a pain. This is probably caused by false interpretation of the stimuli due to its disturbed processing in the dorsal root ganglia; 5) a contribution of the cerebral cortex in the perception and processing of the pain, as a part of complex, homeostatic matrix; 6) a role of sympathetic pain component in Complex Regional Pain Syndrome. Afore-mentioned hypotheses do not allow to definitive identification of the major patophysiological mechanism which contribute in the development and maintaining of pain in CRPS, but they show a complexity of these processes. PMID- 19999622 TI - [Measurement of ventricular twist by 2D speckle tracking early after acute myocardial infarction. Comparison between anterior and inferior infarction]. AB - INTRODUCTION: Left Ventricular twist (LV twist) is defined as the apical counter clockwise rotation relative to the clockwise basal rotation. It has been shown that LV twist decreases after myocardial infarction (MI) and that it is well correlated with left ventricular ejection fraction. Most studies have only evaluated anterior wall MI. The aim of our study was to determine whether LV twist is dependent on the infarct territory (anterior vs. inferior) and whether there is a correlation between LV twist and matrix metalloproteinase-9, a marker of LV remodeling. METHODS: We measured LV twist using echocardiography with 2D speckle tracking in patients with acute MI and in a control group. RESULTS: We evaluated 27 controls and 35 patients with acute MI, 15 with anterior wall and 20 with inferior wall MI. LV twist was significantly decreased after MI, compared to the control group (10.93 +/- 2.05 vs 15.5 +/- 2.29; p = 0.003). There was no difference between anterior and inferior MI. LV rotation was decreased in the infarct area. We did not observe a correlation between LV twist and MMP-9, or creatine phosphokinase. CONCLUSION: With this study we confirm that LV twist decreases after acute MI. Moreover, we show that LV apical rotation is mostly decreased after large anterior MI. As apical rotation is important for ejection and aspiration (untwisting), this could be a possible mechanism of LV dysfunction after MI. PMID- 19999623 TI - Increase in prevalence of allergy-like respiratory symptoms in IDDM patients contrasting with a decrease in positive skin prick tests. AB - AIM: The goal of this study was to investigate a possible relationship between allergic symptoms, atopy skin prick test results in type 1 diabetic patients and in non diabetic controls. METHODS: 50 type 1 diabetic patients and 50 healthy control subjects were evaluated for allergic disorders with regard to medical history. All persons were skin-prick-tested for atopic sensitization with a common panel of inhalant and food allergens. RESULTS: Although in the diabetic group 56% of the patients reported allergy-like respiratory symptoms, only 22% were found to have positive skin tests. In the control group 22% had allergic complaints and skin prick tests were positive in 34% of the cases. CONCLUSION: Nasal and bronchial allergy-like symptoms were increased in IDDM patients in comparison to a control population. However there was a small decrease (not significant) in prevalence of positive atopy skin tests in diabetic patients in comparison to controls. This suggests that IDDM patients have an important load of allergy-like symptoms that are not linked to an IgE sensitization. PMID- 19999624 TI - Results of the ALEGRIA study in Luxembourg. An epidemiological, observational study to describe symptom impact and control in patients with GERD and an evaluation of the GERD Impact Scale. AB - BACKGROUND AND STUDY AIMS: The aim of this observational study was to perform the first epidemiology study in a primary care patient population with GERD in the Grand Duchy of Luxembourg and to evaluate the added value of the GERD Impact Scale (GIS) patient questionnaire. PATIENTS AND METHODS: 152 Patients with symptoms of GERD from 20 study centers were included. At visit 1, demographic data including lifestyle factors and the patients' symptoms were recorded. GERD symptoms and their severity, treatment changes and the GIS were all assessed at baseline (visit 1), visit 2 (4-6 weeks) and visit 3 (8-14 weeks). Analyses were performed on an intent-to-treat basis. RESULTS: 142 patients were included in the analysis, which comprised 50% men and 50% women with a mean BMI of 27 kg/m2. Documented lifestyle factors included consumption of caffeine-containing beverages (87% of patients), stress (62%) and alcohol consumption (53%); 44% of patients were smokers or ex-smokers. The median duration of GERD was 2.0 years. Upon inclusion, 46% were receiving, or had received, proton pump inhibitors (PPIs), antacids (44%), H2-receptor antagonists (21%) or no treatment (21%). PPIs were prescribed at the first visit in the majority of cases (94%) with 75% of patients being prescribed esomeprazole with a median daily dose of 40 mg. The GIS score correlated well with the clinician's judgment of symptom severity and was reported to help determine the appropriate treatment and evaluate the patient's response in approximately 80% of patients. CONCLUSIONS: In this, the first epidemiological study on GERD patients in the Grand Duchy of Luxembourg, data was obtained as planned. The novel patient questionnaire was judged to be helpful by the physician and data shows that the GIS may have an added value over current assessments. PMID- 19999625 TI - [What can the physician do when faced with the gigantic puzzle of abuse?]. AB - The entire subject of abuse can be compared to a gigantic puzzle were patients as well as their physicians get lost. The goal of this article is first to find a common definition to all acts of abuse and then to subdivide them into three distinct categories: sexual, physical, and emotional abuse. The treating physician should realize that victims of abuse, after the passage of time, will eventually fall sick and consult a doctor for physical or/and psychological symptoms which are the result of the past abuse. Most of the time, these patients will not speak spontaneously of the abuse, either because they have not understood the cause/effect relation, or because the abuse feels unspeakable. It is therefore necessary for the doctor to know how to detect abuse through symptomatology, how to help these patients reveal their secret, how to listen with a non-judgemental ear, and how to propose appropriate treatment. PMID- 19999626 TI - [Telemedicine assisted diet and diagnosis management in food hypersensitivity]. AB - Several studies have shown that the current prevalence of food allergy in Western Europe is about 4% and will further increase. Because of missing therapeutic alternatives, food allergy patients are required to identify individual allergens in the daily food by carefully reading the product's ingredient lists. Experiences with other chronic diseases show that a combination of telemedicine and Disease Management (DM) could have a positive impact on medical outcome and health related costs. A new concept of telemedicine support for allergy patients and allergists has been elaborated within the Luxembourgian MENSSANA project. Instead of measuring physiological parameters, a Smartphone based Personal Allergy Assistant (PAA) allows patients to keep an electronic patient diary by scanning the barcode of the consumed food products. For diagnostic purpose, the diary is regularly transmitted to the allergist's electronic patient record. To further support the individual diet management, the PAA gives a warning before consumption of allergenic food. Computer readable food ingredient lists are required for the PAA diet management. To collect this kind of information, a dedicated web-based "virtual community" of food consumers and producers (www.wikifood.eu) has been established. This volunteer network complements an independent product database. Up to now, more than 13.000 food descriptions are public available within wikifood.eu. PMID- 19999627 TI - Wrong-site surgeries: will non-coverage by CMS produce adverse results? PMID- 19999628 TI - Stimulus bill funds overdue changes to U.S. health care technology. PMID- 19999630 TI - [Information on the fascia that is necessary for rectal surgery: with special reference to its extension]. PMID- 19999629 TI - [Recent topics on epidemiology of colonic cancer]. PMID- 19999631 TI - [Significance of chemo-radiotherapy of rectal neoplasms]. PMID- 19999632 TI - [The current status of adjuvant chemotherapy of colonic cancer]. PMID- 19999633 TI - Daughters' stories: family memory and generational amnesia. AB - After World War II, most Bulgarian Jews emigrated legally to Israel. Those who stayed had to take part in the building of socialism and integrate in a monolithic "socialist nation." Thereby they had to "forget" their ethnic identity ("aided by the state in various ways) and to become "Homo politicus" rather than "Homo ethnicus." Since 1990, a revival of Jewish identity has begun in Bulgaria. Here I explore how the women of three generations from the same family reinvent their Jewish identity in their life stories. Drawing on this particular case, I suggest an approach to the question of the interplay of individual and collective memory. I focus on family and generation as different types of collectivities influencing individual memories and self-actualizations. PMID- 19999634 TI - Recalling war trauma of the Pacific War and the Japanese occupation in the oral history of Malaysia and Singapore. AB - The Pacific War and the Japanese Occupation were traumatic periods in the lives of people now over seventy years old in Malaysia and Singapore. This study traces why individuals interviewed for oral history of the Pacific War and the Japanese Occupation have often been able to tell stories of trauma without being overwhelmed by their reminiscences. It emphasizes that memories of traumatic experiences of the Pacific War and the Japanese Occupation in Malaysia and Singapore are mediated and eased by supportive social networks that are part of the interview subject's community. The individual's personal memories of traumatic war experiences are positioned in the context of the collective memory of the group and, thus, are made easier to recall. However, for individuals whose personal memories are at variance with the collective memory of the group they belong to, recalling traumatic experiences is more difficult and alienating as they do not have the support in their community. The act of recalling traumatic memories in the context of the collective memory of a group is particularly relevant in Malaysia and Singapore. These countries have a long history of being plural societies, where although the major ethnic groups -- the Malays, Chinese, and Indians -- have lived side by side peacefully, they have lived in culturally and socially separate worlds, not interacting much with the other groups. The self -- identity of many older people who lived through the Pacific War and the Japanese Occupation is inextricably bound up with their ethnicity. Oral history on war trauma strongly reflects these identities. PMID- 19999636 TI - Prophecy, patriarchy, and violence in the early modern household: the revelations of Anne Wentworth. AB - In 1676 the apostate Baptist prophet Anne Wentworth (1629/30-1693?) published "A True Account of Anne Wentworths Being Cruelly, Unjustly, and Unchristianly Dealt with by Some of Those People called Anabaptists," the first in a series of pamphlets that would continue to the end of the decade. Orignially a member of a London Baptist church, Wentworth left the congregation and eventually her own home after her husband used physical force to stop her writing and prophesying. Yet Wentworth persisted in her "revelations." These prophecies increasingly focused on her response to those who were trying to stop her efforts, especially within her own household. This article examines Wentworth's writings as an effort by an early modern woman, using arguments of spiritual agency, to assert ideas about proper gender roles and household responsibilities to denounce her husband and rebut those who criticized and attempted to suppress her. PMID- 19999637 TI - Criminalizing knowledge: the perverse implications of the intended use regulations of off-label promotion prosecutions. AB - Your company has spent months designing a compliance program and training your sales representatives. They know never to mention the off-label uses of your product. If they are asked about the off-label uses by the physician they are detailing, they know to forward those inquiries to the scientific liaisons at headquarters. But, could your company still be in legal jeopardy simply because it knows that the product is being used for an off-label purpose? This article attempts to track the Food and Drug Administration's (FDA's) shifting interpretation of its "intended use" regulations, from focusing entirely on the statements of the manufacturers to focusing on the knowledge of the industry, indeed, of the consumers of products, in determining the true intended use of a product. It will look at several recent attempts by FDA to use that new interpretation of the regulations to expand its power: to regulate tobacco and to require pediatric indications for any new drug. Finally, it will look at several recent examples of how this new interpretation has manifested in actions by FDA and the Department of Justice (DOJ). PMID- 19999638 TI - Tracing sexual identities in "old age": gender and seniority in advice literature of the early-modern and modern periods. AB - Thus far, historians have interpreted representations of elderly women with reference to women's roles or to women's positions in society. This article proposes a different approach toward gender: to relate representations of the aged to the sexual identities of both men and women. This article analyzes representations of old age in conduct books of the early-modern period and the nineteenth century. By drawing a comparison, the eighteenth-century change of "identity regime" in European culture is brought to the fore. The article points to the influence of sexual identities on the representations of senior persons in advice literature both in Dutch and translated into Dutch. PMID- 19999639 TI - FDA regulation of tobacco: blessing or curse for FDA professionals? AB - Upwards of 400,000 Americans will die that year from the effects of cigarettes, which FDA will now "regulate" very gently, with its hands tied by a slick statutory protection for the largest existing tobacco marketers. Career FDA professionals will be criticized as enablers of mega-marketers' continued sales, working at the margins, arranging the paperwork for protection of megafirms' market share, and sitting by as the deaths and addictive behaviors continue. "Join the Public Health Service, inspired by a public health mission," they were told, and yet they will be unable to do much regulating of the addictive and fatal products for which they now have titular responsibility. This essay observes that these fine FDA professionals are handed the sticky remains of a messy bargain, negotiated in a distracted Congress by expensive lawyers with clients who were potent contributors to political action committees. The only formula that is not secret about the 2009 law is the way in which industry purchased sufficient allegiance to gather the votes for its adoption. The remaining mystery is how FDA could be expected to do these tasks without losing its best and brightest professionals to other fields. PMID- 19999640 TI - Cloned animal products in the human food chain: FDA should protect American consumers. AB - Animal cloning is "complex process that lets one exactly copy the genetic, or inherited, traits of an animal." In 1997, Dolly the sheep was the first animal cloned and since then "scientists have used animal cloning to breed dairy cows, beef cattle, poultry, hogs and other species of livestock." Cloned animals are highly attractive to livestock breeders because "cloning essentially produces an identical copy of an animal with superior traits." The main purpose of cloning livestock is "more focused on efficiency and economic benefits of the producer rather than the overall effect of cloning on an animal's physical and mental welfare." The focus of this article is threefold. First, the science behind animal cloning is explained and some potential uses and risks of this technology are explored. Second, FDA's historical evolution, current regulatory authority, and limitations of that authority, is described. Lastly, a new regulatory vision recognizes the realities of 21st century global markets and the dynamic evolution of scientific discovery and technology. PMID- 19999641 TI - Dietary supplement adverse event reports: review and analysis. AB - The Food and Drug Administration (FDA) works actively to ensure that all the dietary supplements currently in the market are safe for the public. Existing policies are often remodeled and new regulations are developed periodically. A recent change to supplement regulations includes the implementation of a system that allows consumers to contact the FDA or product manufacturer directly to report an adverse reaction as a result of dietary supplement consumption. FDA has employed an adverse event reporting (AER) system that is aimed to police postmarketing effects of dietary supplements.This article will shed light on the importance of tracking serious adverse events with respect to dietary supplements that have already been marketed. Using this data, dietary supplement companies will be able to take the necessary steps to ensure that their products remain safe for the public by removing problematic ingredients from their products and engaging in more effective labeling and product disclaimer practices. PMID- 19999642 TI - Encouraging research and development of pediatric medical devices through legislative and regulatory action: the Pediatric Medical Device Safety and Improvement Act of 2007 in context. AB - In September 2007, Congress passed the Food and Drug Administration Amendments Act of 2007 (FDAAA). Title III of the FDAAA, the Pediatric Medical Safety and Improvement Act (Title III), created new incentives, mandates, Food and Drug Administration (FDA) authority and funding with the aim of increasing the availability of devices for pediatric populations while assuring the safety and effectiveness of those devices. This article describes the complex problem Title III addresses and situates Title III within the context of the regulatory scheme previously in place, particularly in relation to the one in place to address the parallel problem for pediatric drugs, and the concerns and policy recommendations of diverse stakeholders voiced leading up to Title III. PMID- 19999643 TI - Status homogamy in the preindustrial marriage market: partner selection according to age, social origin, and place of birth in nineteenth-century rural Sweden. AB - This article studies partner selection according to three dimensions: social origin, age, and place of birth. The authors use micro-level data from local population registers in five parishes in southern Sweden from 1815 to 1895. The results confirm that all three aspects were important but that socioeconomic status was the most important characteristic, structuring much of the selection process. The importance of social and age homogamy remained stable over the period, while geographic exogamy became more frequent, which could be interpreted in terms of an increasing openness of rural society. The authors also find some indications of exchange of characteristics in the partner selection process. PMID- 19999644 TI - Obesity, fast food manufacture, and regulation: revisiting opportunities for reform. AB - Regulations have historically been able to shape public behavior in various ways. As poor dietary practices and obesity continue to pose major health and economic threats to society, attention will continue to be directed towards the ethical and legal responsibilities of fast food manufacturers as potential contributors to these problems. In light of these considerations, several opportunities emerge that may impact dietary behavior and obesity through regulation of the fast food industry. This article addresses the health consequences of fast food consumption, as well as the historical and legal contexts of fast food regulation in the United States. PMID- 19999645 TI - Opacity and cost effectiveness analysis in Medicare coverage decisions: health policy encounters administrative law. AB - Medicare is the public health insurance program for elderly and disabled Americans. Established in 1965, today it is the primary health insurer for more than 40 million people. Like all insurance programs, Medicare must determine what benefits to cover. In the Medicare statute, Congress established coverage for broad categories of care--hospitalization, office visits and prescription drugs- and broad categories that were excluded, such as vision and dental care. Beyond these broad categories, Congress left the agency administering Medicare--now known as the Center for Medicare and Medicaid Services (CMS)--to determine whether certain treatments are "reasonable and necessary" and thus eligible for Medicare coverage.Two "facts" about Medicare coverage determinations are well established in the health policy literature. First, CMS does not have criteria by which it makes coverage decisions. Second, CMS does not conduct cost effectiveness analysis in making these determinations. Although CMS approves coverage of most treatments it is asked to evaluate--99 percent of the time in one study--it does deny coverage of some procedures and devices. Therefore, some criteria for denying coverage must exist, even if they are not published or otherwise formalized. This article demonstrates that, as both a legal and policy matter, these explanations are inadequate. PMID- 19999646 TI - FDA's expanding postmarket authority to monitor and publicize food and consumer health product risks: the need for procedural safeguards to reduce "transparency" policy harms in the post-9/11 regulatory environment. AB - This article provides a summary of the expansion of FDA's discretionary authority in the post-9/11 period, particularly with respect to FDA's authority to monitor and publicize potential health risks linked to food, dietary supplements, nonprescription drugs, and other consumer health products. In addition, this article evaluates the need for FDA to establish procedural safeguards to reduce the significant risks of unintended and undue harm to people and regulated companies that can result from adverse publicity in the more "transparent" post 9/11 FDA regulatory environment. Specifically, Part I summarizes the amendments to the FDCA enacted during the post-9/11 period that have expanded FDA's postmarket authority to monitor, evaluate, and publicize potential health risks linked to food, dietary supplements, nonprescription drugs and other consumer health products marketed in the United States, in conjunction with FDA's Sentinel Initiative, Reportable Food Registry, and other adverse event reporting requirements. Part II discusses the convergence of FDA's expanded postmarket authority to publicize product-related risks with President Obama's transparency initiative aimed at fostering "open government" through increased public access to government information. In addition, Part II considers the nature of the procedural safeguards needed in the post-9/11 FDA regulatory environment, in view of FDA's historical record and illustrative cases that help expose how adverse "transparency" surrounding FDA warning letters, recalls and safety alerts concerning products in the marketplace can have undue and unintended prejudicial and harmful effects for the people and companies that are legally responsible for such products. Finally, based on these analysis, this article concludes with some observations concerning the nature of the procedural safeguards needed to reduce the significant risks of "transparency" policy harms in the pos-9/11 regulatory environment. PMID- 19999647 TI - Nursing practice statement NP 02: Guidelines for medication administration in schools. Developed: 9/97 * revised: 7/2009. PMID- 19999648 TI - H1N1 vaccination--the implications for RNs. PMID- 19999649 TI - Validating tumor linkage using the NAACCR site pairs table. AB - The objective of this study is to illustrate use of the Site Pairs Table developed by the North American Association of Central Cancer Registries (NAACCR) Record Linkage Work Group to validate tumor linkage in a central registry database and to identify potential cases with inaccurate tumor linkage. Central registries often receive reports for patients with multiple tumors, and they receive multiple reports from different sources for the same tumor. Tumor site pairs (pairs of unique tumors for patients with multiple tumors) ought not refer to the same tumor as represented in the Site Pairs Table. Likewise, abstract pairs (pairs of abstracts relating to the same tumor) ought to be identified during the tumor linkage process as belonging to the same tumor. Three central cancer registries represented on the work group contributed data to the study. The data included cases diagnosed 1992-2003 and represented 143,288 patients with multiple tumors and 280,227 tumors with multiple abstracts. Totals of 181,118 tumor site pairs and 391,670 abstract site pairs were generated from the data and compared to the Site Pairs Table. Of the abstract site pairs 381,389 (97.4%) were found in the Site Pairs Table. One registry reviewed its portion of the 2.6% not found in the table and determined 12% of the cases were incorrectly linked and should change from one tumor to two tumors. Of the tumor site pairs, 144,793 (80%) were not found in the Site Pairs Table. Further evaluation of the remaining 20% by paired site and laterality, histology and timing showed 19.3% were considered unique tumors and 0.7% were identified as potential cases with inaccurate tumor linkage. Two registries reviewed their portion of these cases. One registry changed two tumors to one tumor on 44% of the cases they reviewed. The other registry changed two tumors to one tumor on 53% of the cases they reviewed. Analyzing site pairs within the registry database using the Site Pairs Table assists in identifying inaccurate tumor linkages as was shown in this study. PMID- 19999650 TI - Patterns of cancer in first generation immigrants from the Arab League and other countries. AB - Arabic immigrants are a relatively new group of immigrants and one of the fastest growing populations in the United States. Yet, information about their disease patterns is limited because Arabic is not a recognized minority group in population statistics. We tested whether population-based cancer registry data were useful to describe cancer patterns of the first generation of Arabic immigrants, compared with non-immigrants as well as the first generation of other immigrants. Information on invasive cancer cases with only one primary and known age was extracted from Surveillance, Epidemiology, and End Results (SEER) public use data (1973-2004) and software. Immigrants were divided by place of birth; Arabic: born in the Arab League countries, and other immigrants: born in countries other than the United States or Arab League, whereas US-born were used as a reference population. Information on place of birth was frequently absent for non-deceased cases, therefore data were limited to deceased cases. Age- and gender-standardized proportional ratios (SPR) were calculated for 24 aggregated cancer sites. Significantly high or low SPRs were observed for Arabic immigrants for the following cancer sites: liver (3.15), gallbladder & other biliary (1.87), thyroid (1.74), stomach (1.54), leukemia (1.41), lymphoma (1.36), pancreas (1.36), ovary (1.29), brain and other nervous system (1.23), lung (0.80), prostate (0.72), oral cavity and pharynx (0.64), esophagus (0.39), and skin melanoma (0.33). These patterns were similar for the other immigrants; however, ovary and brain were Arabic-specific, and thus may deserve further investigation for specific environmental or genetic exposures among Arabic immigrants. PMID- 19999651 TI - Obtaining information on language preference among newly diagnosed Hispanic and Asian American cancer patients in Connecticut. AB - Information on primary or preferred language in hospital records of the growing numbers of newly diagnosed Hispanic and Asian American cancer patients could be useful in studies of quality of cancer care and prognosis. Using the statewide Connecticut cancer registry and medical records at acute-care hospitals in Connecticut, only 9.7% of 992 Hispanic or Asian American patients had no information on language preference. Information on use of an interpreter was missing for 36.1% of 653 probable non-English-preferring patients. Missing information was more frequent in Asian than Hispanic American patients. The potential for adding this information to national databases should be explored. PMID- 19999652 TI - Going the distance for certified cancer registrars. AB - Cancer registry departments are using electronic technology to solve the local and national Certified Tumor Registrar (CTR) shortages. As demand for CTRs continues to increase without an accompanied increase in the supply of qualified personnel, cancer registry departments are looking for new solutions to this growing local and national trend. In order to solve this problem, some cancer registries have started using telecommunication to fill the empty positions within their departments. This is the case at Roper St. Francis Healthcare (RSFH) in Charleston, SC, where Cancer Registry Manager, Ellen Kolender, RHIA, CTR, used telecommuting to fill one full-time and one part-time CTR position. PMID- 19999653 TI - Grasping the essentials of CP3R. PMID- 19999655 TI - The ledge. PMID- 19999656 TI - 50 years of staging. PMID- 19999657 TI - [Psychiatric out-patient care]. AB - There is a dearth of solid scientific data on the effectiveness of regular psychiatric outpatient care. Models for psychiatric community treatment rose from pioneering work, but these models have been tested in a reletively small number of high quality randomized and controlled trials. Some of psychiatric hospital care could be replaced by day-hospital care and use of hospital treatment may be decreased among some of the patients with severe psychiatric disorders by using home-based care by mental health teams. Early intervention in psychosis may provide possibilities to decrease need of hospital care. Closure of psychiatric hospitals is unlikely to lead to the development of community care. High quality care necessitates follow-up of effectiveness and, additionally, good working conditions, where treatment may be applied according to the best information available. PMID- 19999658 TI - [Cardiac sarcoidosis]. AB - Cardiac sarcoidosis is a severe inflammatory disease of the cardiac muscle, manifesting itself as atrioventricular block, ventricular tachycardias, cardiac insufficiency and combinations thereof. Approximately half of cardiac sarcoidosis patients exhibit no clinical signs of sarcoidosis outside the heart. The diagnosis is based on cardiac muscle imaging and myocardial biopsy. High dose corticosteroid medication is utilized for treatment. A life-threatening cardiac event occurred in more than one third of cardiac sarcoidosis patients at Meilahti hospital. PMID- 19999659 TI - [Supporting a patient's self-management]. AB - Regardless of favorable results of health care, citizens still face with health risks which are associated with the life style. Shift from professional-centered care to patient-centered care sets challenging tasks and roles to professionals as well as patients. Professional's authority is transformed into a role of a patient's partner and trainer, with whom the patient negotiates the best treatment for him or her. Professional taking the role of a partner and a trainer strengthens the patient's self-management skills by supporting patient's empowerment, autonomy, motivation and self-efficacy when the patient is solving his or her health problems and making decisions. From the patient's point of view, self-management demands knowledge of objectives and means of treatment as well as willingness to take responsibility for one's own treatment and ways of living, as well as confidence to one's own resources and capabilities. PMID- 19999660 TI - [Identification of susceptibility genes for age-related macular degeneration--a success story of molecular genetics]. AB - The number of persons over 70 years of age with advanced age-related macular degeneration in Finland can be estimated to be approximately 50,000. Milder forms are additionally present in a considerably larger group. Smoking and age are undisputed non-genetic risk factors of age-related macular degeneration. Of the genetic factors, polymorphisms of the complement factor H (CFH) and LOC387715 genes have a strong impact on the risk of developing the disease, whereas alleles of the C3, CFB, and the C1 inhibitor SERPING1 genes of the complement system exhibit only minor effects. PMID- 19999661 TI - [Competence in the medical profession: a need for a wider perspective?]. AB - Working as a physician requires versatile competence, which has been dissected by using various models. Most of these include medical knowledge, patient work, interactive skills, multiprofessional collaboration, management of operational processes within medical and health care, and information technology skills. Without feedback, the ability of physicians to evaluate their developmental needs is poor, whereby the development of competence should be tied on the basis of needs to on-the-job learning and functionality. Different perspectives should be used in its planning and evaluation. After the evaluation, a development plan should be devised and resources created for its implementation. PMID- 19999662 TI - [An infant less than three months of age at emergency call service]. AB - Examination and treatment of an infant less than three months of age requires different action at the emergency department as compared with bigger children. Even slight cough and vomiting may be indications of a severe disease in this age group. In addition, symptoms in small infants are often diffuse and sometimes difficult to observe and analyze. A general assessment by the physician constitutes the basis for diagnostics. Of the severe diseases, most are infections. PMID- 19999663 TI - [Complications associated with inguinal hernia repairs at the Patient Insurance Centre in 2003 to 2007]. AB - BACKGROUND: The aim was to elucidate the prevalence of complications associated with adults' inguinal hernia repairs in Finland. MATERIALS AND METHODS: We reviewed the notices of patient injury associated with inguinal hernia repairs, as reported in 2003 to 2007 to the Patient Insurance Centre. We analyzed the associated causes, compensations, and compensation criteria. The numbers and types of hernia operations performed during the same period in the whole country were retrieved from the notification register. RESULTS: A total of 55,000 hernia operations were carried out during this period, resulting in 250 notices of patient injury. Of these, 92 resulted in compensation. CONCLUSIONS: Besides severe pain, serious complications from inguinal hernia repairs seem to be rare. PMID- 19999664 TI - [Chemoembolization of hepatocellular carcinoma with drug eluting beads]. AB - Rates of hepatocellular carcinoma are increasing worldwide, and most patients can only be offered palliative treatment. Recent advancement is chemoembolization with drug eluting beads (DEB, Biocompatibles). We treated 12 patients with DEB. Tumour response was evaluated by computer tomography two months after each procedure and clinical data and survival of patients was collected from case reports. Tumour necrosis was found in nine, but not in three patients. Eight of twelve patients were alive after one year. Severe procedure-related complications were not seen. Our primary experience is that chemoembolization with drug eluting beads is an effective and safe procedure for treatment of hepatocellular carcinoma. PMID- 19999666 TI - [Land wine of grand cru]. PMID- 19999665 TI - [Update on current care guidelines. Treatment of severe sepsis in adults]. AB - According to the Finnsepsis Study, the incidence in Finland of severe sepsis requiring intensive care was 0.38/1,000 inhabitants/year. ICU and hospital mortality was 15.5% and 28.3%, respectively. The Finnsepsis Study showed that compliance with protocols was rather poor and antimicrobial treatment was often delayed. These guidelines emphasize the importance of prompt antibiotic and fluid therapy. In shock, norepinephrine is the first line vasopressor. Low-dose hydrocortisone may be used to shorten the need for vasopressors. Activated protein C should be considered in selected patients. The blood glucose target recommendation is between 5 and 8 mmol/l. PMID- 19999667 TI - [Oral health care is not a trade]. PMID- 19999668 TI - [Removable partial dentures. Oral functions and types]. AB - A removable partial denture enables the restoration or improvement of 4 oral functions: aesthetics, mandibular stability, mastication, and speech. However, wearing a removable partial denture should not cause oral comfort to deteriorate. There are 3 types of removable partial dentures: acrylic tissue-supported dentures, dentures with cast metal frameworks en dentures with cast metal frameworks and (semi)precision attachments. Interrupted tooth arches,free-ending tooth arches, and a combination of interrupted as well as free-ending tooth arches can be restored using these dentures. Well-known disadvantages of removable partial dentures are problematic oral hygiene, negative influence on the remaining dentition and limited oral comfort. Due to the advanced possibilities of fixed tooth- or implant-supported partial dentures, whether or not free-ending, or tooth- as well as implant-supported partial dentures, the indication of removable partial dentures is restricted. Nevertheless, for the time being the demand for removable partial dentures is expected to continue. PMID- 19999669 TI - [Indications for removable partial dentures]. AB - Since there are many ways of preserving a natural dentition, if necessary with support of solitary crowns and fixed partial dentures, sometimes on dental implants, removable partial dentures are nowadays primarily indicated in patients with complaints about missing teeth in the aesthetic zone, which cannot be solved in another way. In addition to this, a removable partial denture is indicated in patients with extremely reduced dentitions or large or multiple edentulous areas, in patients with severe periodontitis or excessive loss of alveolar bone, in patients who are physically or emotionally vulnerable, as an interim solution on the way to edentulousness, as a temporary solution waiting for more extensive treatment and for patients who cannot afford an alternative. PMID- 19999670 TI - [Treatment planning in patients with a severe reduced dentition]. AB - Because loss of teeth many people have a reduced maxillary and mandibular dentition. The appropriate prosthetic rehabilitation is dependent on whether or not a standard mutilation of the dentition can be achieved. In this article, the focus is on treatment of a remaining mandibular dentition combined with an edentulous maxilla, whether or not with a reduced residual alveolar ridge. In the literature, there is no consensus on the appropriate prosthetic treatment if implant-supported overdentures are not applicable. The amount of bone loss in already during some time edentulous maxillary and mandibular alveolar ridge segments as well as the available vertical space for the removable (partial) denture are important decision-making factors. PMID- 19999671 TI - [Extension of (extremely) shortened dental arches by fixed or removable partial dentures]. AB - Whether a shortened dental arch needs to be extended depends on the degree of the shortening. Four categories of shortened dental arches can be distinguished: 1. slightly shortened dental arches; 2. moderately shortened dental arches; 3. extremely shortened dental arches; and 4. asymmetrical extremely shortened dental arches. Decision-making on extending a shortened dental arch should be based on oral function and the patient's perceived impact on oral health-related quality of life. According to this principle, there is no evidence for extending slightly shortened dental arches. Extending moderately shortened dental arches is indicated in exceptional cases, especially for aesthetic reasons. In the case of (asymmetrical) extremely shortened dental arches there are reasons for extension. The extension can be restricted to restoring the arch to the level of a moderately shortened dental arch. With this treatment strategy, it is possible in many cases to conform to the current trend of using fixed partial dentures rather than removable partial dentures. PMID- 19999672 TI - [Prevention: the success of a partial denture]. AB - After the placement of removable partial dentures, additional caries and periodontal problems may arise in the remaining dentition. These problems are the result of insufficient oral hygiene. Oral hygiene may be hampered by wearing the removable partial denture. The design of the removable partial denture should be carried out according to modern concepts of preventive dentistry. Before placing a removable partial denture, the patient's mouth should be thoroughly healthy and the patient should be instructed properly on adequate oral care. Furthermore, the patient should be aware of his/her own responsibility for maintenance. If cleaning twice a day with fluoride tooth-paste provides insufficient protection, care can be supported by daily use of a fluoride containing antimicrobial mouthrinse. Little effect can be expected from professionally applied fluoride or chlorhexidine solutions. PMID- 19999673 TI - A six-month prospective evaluation of personality traits, psychiatric symptoms and quality of life in ayahuasca-naive subjects. AB - The authors assessed 23 subjects immediately before and six months (27.5 weeks) after their first ayahuasca experience in an urban Brazilian religious setting, either Santo Daime (N = 15) or Uniao do Vegetal (N = 8). Measures included scores on instruments assessing psychiatric symptoms, personality variables and quality of life. Independent variables were the frequency of ayahuasca use throughout the period and the length of ayahuasca wash-out after six months. Santo Daime subjects had a significant reduction of minor psychiatric symptoms, improvement of mental health, and a change in attitude towards more confidence and optimism. The Uniao do Vegetal group had a significant decrease in physical pain, and attitude change towards more independence. Independence was positively correlated with the frequency of ayahuasca use and negatively correlated with the wash-out period. We discuss possible mechanisms by which these changes may occur and suggest areas for future research. PMID- 19999674 TI - Subjective effects of Salvia divinorum: LSD- or marijuana-like? AB - Salvia divinorum is a naturally occurring psychedelic considered to be one of the most potent hallucinogens found to date. The few behavioral studies conducted conclude that Salvia's effects may be similar to traditional psychedelics, which is noteworthy because Salvia acts via a unique molecular mechanism as a kappa opioid receptor agonist. One hundred and ninety-three participants, including 34 Salvia users, were asked to fill out a series of questionnaires related to general drug use, personality characteristics, demographics and their experiences with Salvia. Salvia users were found to differ from nonusers on personality characteristics and reported consuming significantly more alcohol than nonusers. In addition, although Salvia users rated their hallucinogenic experiences as similar to those seen in previously published reports, the majority likened their experiences as most similar to marijuana instead of more traditional psychedelics. Low scores on the ARCI LSD subscale confirmed this finding and call into question the reigning theory of LSD-like subjective effects elicited by Salvia. PMID- 19999676 TI - Methamphetamine in three small Midwestern cities: evidence of a moral panic. AB - This study examined the coverage of methamphetamine from 1997 to 2005 by three newspapers serving small Midwestern cities and contrasted their portrayals of methamphetamine problems with available data on the severity of the meth problem in each locality. Results of quantitative and qualitative content analyses show that--to varying degrees, across sites and over time--newspaper coverage of meth was disproportionate to the scale of the meth problem as indicated by site specific treatment admissions data. To some extent, each of the three newspapers used drug-scare rhetoric (e.g., medical metaphors such as "plague") to describe the prevalence and effects of meth. Results indicate that two of the three newspapers' portrayals of meth were conducive to promoting a moral panic over the drug. Potential explanations for variations in coverage are discussed, and findings are considered in light of research on prior drug scares. PMID- 19999675 TI - Making connections: New Orleans Evacuees' experiences in obtaining drugs. AB - Between August 29 and September 7, 2005, almost all New Orleans residents were evacuated from the area in the aftermath of Hurricane Katrina. News reports indicate that almost 130,000 New Orleans Evacuees (NOEs) were evacuated to Houston, Texas, the largest recipient of the civilian population from New Orleans. Some of these NOEs were active participants in the illicit drug market in New Orleans prior to the hurricane. The period between the flooding and the nearly complete evacuation of New Orleans as well as their subsequent displacement to Houston and other locations provided unique opportunities to study what occurs when illicit drug markets are disrupted, since populations of illicit drug users and purchasers could no longer routinely obtain their drugs in predictable ways. Utilizing qualitative data from in-depth interviews and focus groups, this article describes the ways NOEs (1) managed their drug acquisition and use following evacuation; (2) located new sources of drugs in Houston and elsewhere by tapping into shared drug culture; and (3) gained access to and learned the argot for drugs in the local drug market in new settings. This report contributes to the nascent literature on disrupted drug markets. PMID- 19999677 TI - Effects of methamphetamine abuse beyond individual users. AB - Since 1997, the use of methamphetamine as a drug of abuse has been widespread in the United States. While several forms of amphetamine are useful in some areas of medicine, methamphetamine as an abused substance is associated with severe and multifaceted consequences. Problems associated with the abuse of amphetamine and its derivatives such as methamphetamine have been well documented. As the manufacture and use of methamphetamine across the United States has increased, the impact of methamphetamine abuse has been felt beyond individual users; families as well as communities can be seriously affected. An increase in child neglect and violence as well as a lack of resources for health care, social services, and law enforcement because of methamphetamine abuse have been reported by many communities. This study examines the historical spread of methamphetamine misuse in the United States and the resulting individual, social, and environmental consequences. A public health perspective on family, community, and social aspects is offered, and ideas for future research and policy changes are explored. PMID- 19999678 TI - Sense of coherence as a stable predictor for methadone maintenance treatment (MMT) outcome. AB - We studied stability of sense of coherence (SOC) over time and in relation to the outcomes of former heroin addicts in MMT. Between September of 2003 and April of 2006, all 90 newly admitted patients to a MMT clinic in Tel Aviv were studied and followed up for one year. A SOC (inner resources) questionnaire was done at baseline and after one year. Use of opiates, cocaine, benzodiazepines, cannabis and amphetamines in the 13 months after admission was recorded, and defined as positive if at least one urine test for any drug was positive. After one year, 77 (85.6%) stayed in treatment. SOC scores were similar at baseline and after one year, but were lower among 42 patients who still abused any drugs (118.8 + 27.4), compared with the 35 patients who did not (128.5 + 26.5, Repeated measures, Groups p = 0.003, Time effect p = 0.5, Time*Group p = 0.003). Cumulative retention revealed that 30 patients with SOC scores greater than 130 had longer retention (3.5 years, 95% CI 3.2-3.9) as compared with 60 patients with SOC scores of 130 or less (2.9 years, 95% CI 2.5-3.3). SOC is a stable parameter that can serve as a predictor for success in MMT, both as retention in treatment and drug abstinence. PMID- 19999679 TI - Gender differences in crime, drug addiction, abstinence, personality characteristics, and negative emotions. AB - The current study examined gender differences in personal and psychological characteristics among drug-abstinent Israeli inmates. The study focuses primarily on three personal variables: sense of coherence (SOC), anxiety, and hostility. Additional factors that were examined are demographic variables, which include background, crime, and drug addiction. The sample included 119 inmates (65 males and 54 females) who abstained from drugs use for two different time intervals--up to one year, and more than one year--and did not receive treatment. Overall, males and females shared similar backgrounds except for a higher rate of sexual abuse among female inmates. No gender differences were found in recidivism. These findings show that drug-addicted female inmates injected drugs to a greater extent than males. They also indicated that male inmates remained abstinent for longer periods than female inmates. Additionally, length of abstinence was related to higher SOC, lower trait anxiety, and less hostility among male inmates compared to female inmates. Among female inmates, length of abstinence was related to lower SOC, higher trait anxiety, and no change in hostility. These findings may indicate the need to require separate gender-oriented therapeutic interventions. PMID- 19999680 TI - Gender differences in psychosocial functioning across substance abuse treatment. AB - The present study investigated gender differences in psychosocial functioning among clients mandated to substance abuse treatment. In the current study, 210 probationers' self reports of depression, anxiety, self esteem, decision-making, life beliefs, role modeling, cooperation, and AIDS risky behavior were gathered. Ratings were collected at intake as well as midway and toward the end of the residential phase of treatment. A series of growth curve models were used to examine changes in psychosocial functioning across these time periods. Improvements were recorded in several areas of psychosocial functioning over the course of treatment (including anxiety, depression, self esteem, decision-making, role modeling, and life beliefs). Importantly, significant gender by time interactions indicated that females showed greater rates of change than their male counterparts in such areas as self esteem, decision-making, and role modeling. The importance of psychosocial change and the implications of these gender differences are discussed. PMID- 19999681 TI - Modified therapeutic communities and adherence to traditional elements. AB - Traditional therapeutic communities (TCs) are characterized by confrontational group therapy, treatment phases, a tenure-based resident hierarchy, and long-term residential care. Many TCs have modified the structure and intensity of the traditional model, tailored services for specific client populations, and hired more professionally trained staff. This study examines the extent to which modified TCs are able to retain the underlying core technology of the TC. Using data from a nationally representative sample of 380 self-identified TCs, six traditional TC elements are identified. Results from a structural equation model indicate that offering services for specific populations and professionalization of staff has limited impact on the six TC elements. Modifications to structure and intensity of TC programming evidenced the strongest effect. Specifically, outpatient-only TCs showed significantly lower adherence to five of the six elements. Short-term residential programs showed a similar negative trend. Findings suggest selected modifications are possible without significantly impacting the TC model's core technology. PMID- 19999684 TI - Availability of websites offering to sell psilocybin spores and psilocybin. AB - This study assesses the availability of websites offering to sell psilocybin spores and psilocybin, a powerful hallucinogen contained in Psilocybe mushrooms. Over a 25-month period beginning in March 2003, eight searches were conducted in Google using the term "psilocybin spores." In each search the first 100 nonsponsored links obtained were scored by two independent raters according to standardized criteria to determine whether they offered to sell psilocybin or psilocybin spores. No attempts were made to procure the products offered for sale in order to ascertain whether the marketed psilocybin was in fact "genuine" or "counterfeit." Of the 800 links examined, 58% led to websites offering to sell psilocybin spores. Additionally, evidence that whole Psilocybe mushrooms are offered for sale online was obtained. Psilocybin and psilocybin spores were found to be widely available for sale over the Internet. Online purchase of psilocybin may facilitate illicit use of this potent psychoactive substance. Additional studies are needed to assess whether websites offering to sell psilocybin and psilocybin spores actually deliver their products as advertised. PMID- 19999682 TI - Premature discharge from methadone treatment: patient perspectives. AB - Longer retention in drug abuse treatment is associated with better patient outcomes, and research indicates the first 12 months of methadone treatment are critical to patient success. Nevertheless, large-scale multisite longitudinal studies over the past three decades indicate that the majority of patients drop out during the first year of methadone treatment. Through an examination of 42 qualitative interviews with patients prematurely discharged from six methadone treatment programs in Baltimore, this study highlights factors patients describe as contributing to their reasons for being discharged within the first 12 months of the treatment. The two most consistent themes are program-related factors and incarceration. The former factors are richly described through patients' words and underscore the ways in which patients' perceptions of control exerted by the program and by the medication and misunderstandings of program structure can lead to premature discharge. Patients' reasons for discharge were compared to counselors' reasons as indicated in discharge summary forms. An analysis of the patterns of agreement and disagreement are presented. Patient-centered program and policy implications are discussed. PMID- 19999685 TI - Bad blood: doctor-nurse behavior problems impact patient care. PMID- 19999683 TI - Investigating the effects of San Francisco's Treatment on Demand Initiative on a publicly-funded substance abuse treatment system: a time series analysis. AB - This study investigated effects of San Francisco's Treatment on Demand Initiative, which was designed to increase substance abuse treatment capacity, on the host treatment system. Secondary data on substance abuse treatment admissions, from 1995 through 2000, were obtained from the Department of Public Health, Community Substance Abuse Services, San Francisco, California. Data on 73,988 admissions were retained and grouped by week of admission. Time series analysis was used to assess the effects of time and treatment on demand (independent variables) on weekly number of admissions, sociodemographic characteristics and types of treatment received (dependent variables). As a function of treatment on demand, the number of weekly new admissions significantly increased. Standard outpatient treatment and comprehensive care admissions constituted greater proportions of admissions after treatment on demand. Persons with a primary heroin, cocaine or alcohol problem constituted greater proportions of admissions, and first-time treatment clients constituted a smaller proportion of admissions. Findings suggest that a capacity expansion initiative can increase system wide admissions and redistribute admissions among modalities towards more comprehensive care treatments. PMID- 19999686 TI - 'Disheartening' developments for disruptive behavior. PMID- 19999687 TI - Early intervention can help prevent disruptive behavior. PMID- 19999688 TI - Strengthening physician-nurse partnerships to improve quality and patient safety. PMID- 19999690 TI - The role of a clinical operations analyst in implementing a successful electronic medical record. PMID- 19999689 TI - Crucial conversations: the most potent force for eliminating disruptive behavior. PMID- 19999691 TI - Measuring clinical performance. PMID- 19999692 TI - The ground floor of an internal medicine residency program. PMID- 19999693 TI - Getting ready for EHR, RHIOs and next-generation co-management agreements. PMID- 19999694 TI - Designing and implementing a hospital DVT prophylaxis program. PMID- 19999695 TI - Outlook is bright for physician executive leadership compensation. PMID- 19999696 TI - Advocacy vs. influence. PMID- 19999697 TI - Passing the torch: the never-ending struggle to sustain civilized society. PMID- 19999698 TI - Optimizing organizational meeting management. PMID- 19999699 TI - Best job fit: manager or individual contributor? PMID- 19999700 TI - The association between parent communication and college freshmen's alcohol use. AB - Using a cross-sectional survey, data were collected from 265 first-year college students to determine if parent-student alcohol communication is associated with college drinking or drinking consequences and if this relationship is mediated by students' parental subjective norms, attitudes toward drinking, and perceived risk. Structural equation modeling was used to test hypotheses. Students whose parents talked with them more about the negative effects of alcohol reported more extensive college drinking (beta = 0.12, p < 0.05). Favorable alcohol attitudes were significantly related to both more extensive college drinking (beta = 0.49, p < 0.05) and more drinking consequences (beta = 0.39, p < 0.05). Lower reported perceived risk was significantly related to more drinking consequences (beta = 0.24,p <0.05). Findings indicate that parental communication regarding the negative effects of alcohol may be ineffective at reducing college drinking or drinking consequences. PMID- 19999701 TI - Comparing entering freshmen's perceptions of campus marijuana and alcohol use to reported use. AB - Use of marijuana and alcohol among current college students (N = 1101) was compared to the perceptions and use of entering freshmen (N = 481) surveyed before the start of classes. Entering freshmen significantly misperceived campus norms for marijuana use, over-estimating that almost every student used in the last 30 days, p <.001. Perceptions of alcohol use were relatively accurate. These discrepancies in perception could account for why 40.5% of entering students perceived the campus atmosphere to be promoting marijuana use, whereas only 16.2% perceived the campus atmosphere to be promoting alcohol use. How these misperceptions of social norms might be influenced by the reputation of the campus-and how this might affect potential applicants and enrolled students' behaviors are discussed. PMID- 19999702 TI - A conceptual framework for studying alcohol intake and blood pressure on historically black college and university campuses. AB - Drinking increases the risk of elevated blood pressure, a risk factor for chronic ailments such as hypertension and cardiovascular disease. The experience of elevated blood pressure in young adulthood may be critical for the development of these diseases later in life. College campuses are venues replete with young adults, and drinking is a popular activity in these settings. Because Historically Black Colleges and Universities (HBCU) produce a large proportion of black college graduates, understanding the social context of drinking among young blacks attending HBCUs is important in understanding the role and characteristics of drinking as a risk factor for the development of elevated blood pressure. This article reviews existing literature on alcohol and blood pressure and proposes a conceptual framework linking socioenvironmental factors, stress, and alcohol consumption at HBCUs in the context of elevated blood pressure among young blacks. Recommendations for future research are also proposed. PMID- 19999703 TI - Influence of skip patterns on item non-response in a substance use survey of 7th to 12th grade students. AB - This retrospective case study assessed the influence of item non-response error on subsequent response to questionnaire items assessing adolescent alcohol and marijuana use. Post-hoc analyses were conducted on survey results obtained from 4,371 7th to 12th grade students in Ohio in 2005. A skip pattern design in a conventional questionnaire appeared to increase non-response error rates at first introduction. In previous cigarette users, these errors were associated with under-reporting of alcohol and marijuana use on subsequent questions. These effects were less prevalent in high-achieving students. Skip patterns can contribute to substantial underestimation of alcohol and marijuana use in adolescents. PMID- 19999704 TI - Cultural factors related to smoking in San Francisco's Irish bars. AB - California's Smoke-Free Workplace Act was extended to include bars in 1998. While the majority of bars in the state have become smoke free, in many bars patrons and staff continue to smoke despite the law. The authors present findings from a study which assessed cultural factors related to continued smoking in bars in the city of San Francisco. In bars serving primarily Irish migrants, tight-knit relations within the local Irish bar community together with a reluctance to be the first Irish bar to ban smoking were found to contribute to continued indoor smoking. The findings illustrate challenges to implementing tobacco control policies within ethnic subpopulations and particularly highlight the importance of considering how cultural dynamics within subpopulations may help or hinder such policies. PMID- 19999705 TI - The influence of race and ethnicity on substance use and negative activity involvement among monoracial and multiracial adolescents of the southwest. AB - This study examined predictors of substance use and negative activity involvement among a diverse sample of European American, African American, Hispanic, Native American, and multiracial early adolescents (n = 749) living in a large urban city in the Southwest United States. This study investigated a broad set of predictor variables that tap sociodemographic, peer, family, community, and school influences. Overall, findings from this study confirm that lifetime substance use remains high among youth of color. Of particular concern is this study's finding that multiracial adolescents are at elevated risk to use substances and engage in negative activities. The implications of this study for understanding how risk factors are influenced by race and other variables on different measures of problem behavior are discussed. PMID- 19999706 TI - Effectiveness of skill-based substance abuse intervention among male adolescents in an Islamic country: case of the Islamic Republic of Iran. AB - The prevalence of substance abuse among adolescents from low- and middle-income countries is increasing drastically and requires immediate intervention. The objective of this longitudinal quasi-experimental panel study was to design and implement a skill-based intervention to prevent and reduce substance use among urban adolescents who attended 2 randomly selected high-schools in Tehran, Iran. One-year post intervention data show that substance abuse, knowledge, attitudes, peer resistance skills, level of self-control, self-efficacy, and perceived susceptibility among intervention group were significantly improved, whereas level of self control and attitudes against substance abuse among the control group deteriorated. To efficiently prevent substance abuse among youth primary preventive interventions should be implemented before onset of substance abuse to improve resistance skills and provide adolescents with information and skills needed to develop anti-drug norms. PMID- 19999707 TI - Advantages of the fixed-angle rotor for the separation of density-labelled from unlabelled proteins by isopycnic equilibrium centrifugation. AB - A comparison has been made between the swinging-buckets rotor and the fixed-angle rotor for density-labelling experiments. The enzymes examined were beta galactosidase (beta-D-galactoside galactohydrolase, EC 3.2.1.23), L-phenylalanine ammonia-lyase (EC 4.3.1.5) and acid phosphatase (orthophosphoric monoester phosphohydrolase, EC 3.1.3.2); it was found that the fixed-angle rotor offered greater separation and resolution and shorter equilibration times compared with the swinging-buckets rotor at the same angular velocity. By exploiting the greater flexibility offered by the fixed-angle rotor it is possible to obtain either a 6-fold improvement in separation or a 6-fold reduction in equilibration time. The significance of these results for density-labelling work is discussed. PMID- 19999708 TI - Alpha1-fetoprotein: separation of two molecular variants by affinity chromatography with concanavalin A-agarose. AB - Alpha1-Fetoprotein present in fetal/newborn rat serum and in hepatoma-bearing human serum has been resolved into two molecular variants by concanavalin A agarose affinity chromatography. The concanavalin A-reactive variant has been purified by gel filtration column chromatography, preparative block electrophoresis and immunoadsorption affinity chromatography. PMID- 19999709 TI - Electrophoretic separation of conformational states of alpha-chymotrypsinogen A at high pressures. AB - When reversible denaturation of chymotrypsinogen is produced at elevated pressures and low temperatures, the transition has been observed to occur very slowly. Electrophoretic separation of the transition mixture at high pressures reveals the presence of two distinct molecular species. This evidence, in conjunction with previously reported spectroscopic data, suggests that a simple two-state model provides a useful description of the transition phenomena at high pressures. PMID- 19999710 TI - [Studies on the primary structure of turnip yellow mosaic virus protein. II. Primary structure of the tryptic peptides of aminoethylated protein]. AB - The sequence of the 7 tryptic peptides obtained from the oxidized protein of turnip yellow mosaic virus have been published in 1970. The other peptides obtained, being insoluble, could not be analysed. The protein was reduced, then alkylated with ethyleneimine and split with trypsin. In this paper we submit the sequence of the 14 peptides obtained, which have been separated on Dowex 50-X4 and Dowex 1-X2. These 14 peptides account for the 189 residues constituting the turnip yellow mosaic virus subunit (mol. wt 20,133). PMID- 19999711 TI - [Studies on the primary structure of turnip yellow mosaic virus protein. III. Primary structure of the chymotryptic peptides of aminoethylated protein. Complete sequence]. AB - The complete sequence of the protein of turnip yellow mosaic virus is given. The isolation of the peptides obtained by cleavage of the aminoethylated protein with chymotrypsin is described. The present study includes the overlapping sequences which allow the correlation of the different tryptic peptides already described. Some hypothesis concerning the secondary and tertiary structure of the protein in the viral particle are also given. PMID- 19999712 TI - Amino acid composition and relationships of lupin and serradella leghaemoglobins. AB - The amino acid compositions of two lupin and two serradella leghaemoglobins were determined by conventional techniques. They are as follows: Lupin component I: Ala20, Arg1, (Asp + Asn)17, (Glu + Gln)17, Gly9, His4, Ile9, Leu15, Lys17, Met1, Phe8, Pro5, Ser12, Thr9, Trp3, Tyr2 and Val17. Lupin component II: Ala26, Arg1, (Asp + Asn)18, (Glu + Gln)20, Gly9, His5, Ile10, Leu16, Lys18, Met1, Phe8, Pro6, Ser12, Thr10, Trp3, Tyr2 and Val18. Serradella component I: Ala32, Arg1, (Asp + Asn)10, (Glu + Gln)18, Gly8, His3, Ile5, Leu5, Lys13, Phe8, Pro4, Ser14, Thr7, Try2, Tyr3 and Val12. Serradella component II: Ala37, Arg1, (Asp + Asn)11, (Glu + Gln)20, Gly9, His3, Ile6, Leu17, Lys15, Phe9, Pro5, Ser16, Thr8, Try2, Tyr3 and Val13. Notable features of these are (a) the absence of cyst(e)ine, which suggests that each is a single polypeptide chain (b) the presence of a methionine residue in the lupin leghaemoglobins, and (c) the high contents of alanine, aspartic acid plus asparagine, glutamic acid plus glutamine, leucine, lysine, serine and valine. The relationships of leghaemoglobins to each other and to other groups of haem-containing proteins have been investigated using a statistical procedure based on amino acid composition. PMID- 19999713 TI - The chain composition of tetanus toxin. AB - Although tetanus toxins from cell and culture filtrate appear indistinguishable by several criteria, only the filtrate toxin can be cleaved into two chains by disulfide scission. These chains approximate molecular weights of 95,000 and 55,000. Determinations of sulfhydryl groups and total half-cystine residues for both the cell and filtrate toxins gave values of approximately six and nine, respectively and in filtrate toxin the half-cystine residues are found evenly distributed between the two chains. PMID- 19999714 TI - Studies on bovine cervical mucin. I. The amino acid composition and N-terminal amino acid of bovine oestrus cervical mucin. AB - Bovine oestrus cervical mucin, isolated by gel filtration on Sepharose 6B, was found to be homogeneous (i) in electrophoresis on cellulose acetate at pH 8.6 after release of sialic acid and in acrylamide-agarose gel at pH 8.7 in the presence of sodium dodecyl sulphate, where not a single compact zone was found after staining for proteins, (ii) by analytical ultracentrifugation and equilibrium centrifugation in a CsCl density gradient; as well as (iii) by immunoelectrophoresis, which revealed two kinds of antibodies against mucin, reacting, respectively, with the primary structure and with determinants depending on the integrity of the glycoprotein structure. Alanine was identified as the N-terminal amino acid of the peptide core, which appeared to consist of a repetition of a 32-35 residue sequence, viz. (Thr)8, (Glu, Pro, Ala, Val)3, (Asp, Leu)2 and (Arg, Cys, Ileu)1. No integral numbers could be assigned to (Ser)3.4, (Gly)2.3 and (Phe)0.6 which were also present. PMID- 19999715 TI - The polypeptide composition of epidermal prekeratin. AB - An alpha-fibrous protein, prekeratin, has been isolated from cow snout epidermis with citrate buffer, pH 2.65. Using acrylamide electrophoresis with 0.1% sodium dodecyl sulfate, prekeratin can be shown to contain three polypeptide chains of different molecular weights. The two faster migrating components are very similar with a mol. wt of about 47,000 while the slower one has a mol. wt of about 58,000. Chromatography on a number of molecular sieve and exchange resins does not separate the components, but use of Sepharose 2B with 0.1 M Tris, pH 9.0, containing 10% propanol gives two peaks of protein. The first and major peak contains all three components while the second has only the two with the faster mobility. The two more rapidly migrating components and the slower one were isolated by acrylamide electrophoresis, and the latter has an amino acid composition more compatible with a non-helical protein. Enzymatic digestion with tosyl-L-phenylalanine chloromethylketone-treated (TPCK-)trypsin shows that the component of mol. wt 58,000 is more susceptible to hydrolysis than the other two. These data suggest that prekeratin is not homogenous in composition and consists of several interacting polypeptide chains. One of these components would appear to be non-helical in structure. PMID- 19999716 TI - Preparation of myosin A from body wall musculature of Ascaris lumbricoides. AB - Myosin A (Ascaris myosin A) was prepared from the body wall musculature of Ascaris lumbricoides var. suis showing a single peak in analytical ultracentrifuge pattern, and its sedimentation coefficient was about 6 S which was close to the values reported for other myosins. The ATPase activities determined in various media were 3-4 times lower than those of rabbit skeletal myosin A. Nevertheless, the maximum level of the superprecipitation of actomyosin was 4 times higher for Ascaris myosin A than for rabbit skeletal myosin A in the presence of 0.01 mM ATP. It appears that Ascaris myosin A is capable of contracting the actomyosin system in a lower concentration of ATP than that for the usual actomyosin system. PMID- 19999717 TI - The role of actin sulfhydryls in actin-myosin interaction. AB - Rabbit white skeletal and dog cardiac actins combined fully with the heavy meromyosins at 4 degrees C and low ionic strength (10.03); the complexes (actin heavy meromyosin) were mostly in a heavy component which settled rapidly during rotor acceleration. Increasing the temperature to 24 degrees C favored formation of the heavy component while increasing the ionic strength to 10.3 decreased formation of the heavy component. Modification of actin with p-mercuribenzoate (PCMB) generally reduced the heavy component and viscosity but did not interfere with actin-heavy meromyosin combination. Modification with iodoacetamide or N ethylmaleimide had similar effects. Addition of ATP reduced viscosity and eliminated the heavy component at levels which partially dissociated actin-heavy meromyosin. Modification of heavy meromyosin with PCMB inhibited combination with actin. The characteristics of interaction of the cardiac proteins varied less with temperature but more with ionic strength than those of skeletal proteins. PMID- 19999718 TI - Anionic and cationic dog trypsin. Isolation and partial characterization. AB - One anionic and one cationic dog trypsin have been identified in dog pancreatic juice and isolated. The dog pancreatic juice contained 1.4 mg/ml of anionic trypsinogen and 1.0 mg/ml of cationic trypsinogen, respectively. The conversion of the trypsinogens to active enzymes is autocatalytic with the formation of active enzymes with higher isoelectric points than the zymogens. Although the amino acid compositions of the two trypsins were similar to those of other mammalian trypsins, a significant difference in content of acidic and basic amino acids, respectively, was found between the two dog trypsins. A high extent of cross-inhibition of the dog trypsins and of bovine trypsin by antibodies to each of the dog trypsins indicated immunological cross-reaction between the various trypsins despite lack of demonstrable cross-precipitation in gels. The calculated s(20,w)0 value for both dog trypsins was 3.3 S. The Km value of anionic trypsin for benzoyl-D,L-arginine-p-nitroanilide' HCl was found to be similar to that of cationic trypsin and to that of bovine cationic trypsin. Soybean trypsin inhibitor, chicken ovomucoid, porcine and bovine secretory inhibitors are potent inhibitors of anionic and cationic dog trypsin. PMID- 19999719 TI - Isolation and characterization of pokeweed mitogen-like phytomitogens from Shoriku, Phytolacca esculenta. AB - From saline extracts of Phytolacca esculenta (shoriku) roots, two phytomitogenes were isolated by salting out with (NH4),SO4 and chromatography on DEAE-cellulose and Sephadex G-100 columns. Both fractions were homogeneous on disc electrophoresis and on immunoelectrophoresis. One of these (Fraction E-2) was shown to be similar to pokeweed mitogen in respect to mol. wt (32,000) and amino acid composition. The other (Fraction E-3) was a protein of 18,000 mol. wt. Both fractions had similar biological activities to pokeweed mitogen in their ability to stimulate pig blood lymphocytes in vitro to incorporate tritiated thymidine, and to induce blastoid transformation. Both fractions contained an unusually large amount of cystine, i.e., 18 half-cystine residues % for Fraction E-2 and 22 residues % for Fraction E-3. Although these mitogens were resistant to deproteinizing procedures such as perchloric acid treatment and Sevag's procedure, the DNA synthesis-stimulating activity was inactivated by digestion with Pronase E and Nagarse, but resistant to trypsin, chymotrypsin, deoxyribonuclease, ribonuclease, lysozyme and neuraminidase. The activity was stable at acidic and neutral pH (4-7) but unstable at alkaline pH. The activity at pH 7.3 was stabilized by the addition of Ca2+ or Mg2+. On the addition of more than 2 mM of Ca2+, precipitation of mitogen occurred. From the above results the molecular basis of the mitogenic activity of shoriku mitogen is discussed. PMID- 19999720 TI - Isolation and characterisation of a progesterone- and testosterone-binding globulin from pregnant guinea pig serum. AB - A progesterone- and testosterone-binding globulin, has been isolated from a pool of pregnant guinea pig serum, using DEAE-Sephadex chromatography, gel filtration on Sephadex G-200 and preparative polyacrylamide gel electrophoresis. This globulin appeared to be heterogeneous on ion exchange chromatography, isoelectric focusing and equilibrium ultracentrifugation, indicating a quarternary structure sensitive to concentration and to ionic environment. In line with the steroid binding serum proteins so far described progesterone-binding globulin is a glycoprotein with a carbohydrate content of 42%. Extrapolated sedimentation and diffusion coefficients were 4.52 S and 5.1.x 10(-7) cm2/s, respectively. A partial specific volume of 0.678 cm3/g has been calculated from the chemical composition. A molecular weight of 82,00 was obtained from equilibrium centrifugation experiments in the presence of o0I1 sodium dodecyl sulfate. The electrophloretic mobility at pH 8.6 corresponds to that of an a,lpha1globulin; isoelectric points were 4.4 and 3.5; Stokes radius, 4.9 nm; frictional ratio, 1.74; extinction coefficient at 280 nm, 7.3. Pure progesterone-binding globulin showed an association constant at 4.0 degrees of 14 x 10(9)M-(1) for progesterone. The affinity for testosterone was lower being 8.2 x 10(7) M(-1). The number of high-affinity binding sites was determined to 1.0 or both steroids. PMID- 19999721 TI - Protamines and other basic proteins from spermatozoa of molluscs. AB - The basic proteins obtained from spermatozoa of different species of the phylum Mollusca have been extracted and fractionated. The amino acid analysis and electrophoretic mobility of these proteins show a considerable variation in the types and relative amounts of the components present in different species. In some case-(Gibbula, Haliotis, Loligo, Octopus) the main components are similar to the protamines found in the salmonid fishes, although they appear to be larger in size (40-80 amino acids) and show significant differences in amino acid composition. In other cases (Mytilus, Chiton) a complex mixture of proteins is present, which including somatic-like histones and proteins intermediate in size and composition betweeln protamines and histones. Other molluscs (Ostrea, Spisula, Patella) also contain proteins intermediate in composition between protamines and histones, but their molecular weight appears to be larger than in histones. In Eledone a complex mixture of proteins containing cystine is obtained, with some components rich in arginine. In most species, somatic-like histones are also present. Their type and relative amount are different in each species. The significance of these results towards an understanding of the evolutionary history of these proteins is discussed. It is suggested that these proteins evolved from histone precursors. PMID- 19999722 TI - Isolation and partial characterization of the myelin basic protein from fetal calf brains. AB - The myelin basic protein has been isolated from brains of fetal calves 30-65 cm crown-rump length. It has been shown to be indistinguishable from the adult bovine encephalitogen (A1 myelin basic protein) by electrophoresis on polyacrylamide gels with and without sodium dodecyl sulfate and by chromatography on CM-cellulose, and Sephadex G-100. The subcellular localization, molecular weight, absorption spectrum, amino acid composition and partial sequence confirm the identity of these two proteins. Changes in the concentration of the myelin basic protein during maturation were compared with histological changes in myelination during maturation. The biochemical and histological studies indicated that the synthesis of the myelin basic protein probably occurs concomitantly with myelination. PMID- 19999723 TI - Differences in protein and glycoprotein patterns between mature and immature neutrophil leukocytes. AB - The post-nuclear particulate fractions of highly enriched mature and immature neutrophil preparations were dissolved in buffer containing sodium dodecyl sulfate and analyzed for protein and glycoprotein components by polyacrylamide gel electrophoresis. The electrophoretic patterns of mature and immature neutrophils showed pronounced differences in both protein and glycoprotein bands. There were 12 prominent protein bands in the patterns of mature neutrophils which also stained for glycoproteins. With one exception, these bands were either greatly reduced in intensity or undetectable at corresponding positions in the patterns of immature cells. The patterns of immature neutrophils revealed three prominent glycoprotein bands, two of which were absent and the third was greatly reduced in intensity at corresponding position in the mature cell patterns. The results indicate that a number of new glycoproteins appear in the later stages of neutrophil maturation, whereas other glycoproteins, present in immature cells, are either lost or greatly reduced in amount. These changes are presumably related to the development of specific cell functions that also appear in later stages of cell maturation. PMID- 19999724 TI - In vitro binding of progesterone to receptors in the human endometrium and the myometrium. AB - High affinity, low capacity progesterone binding receptors have been identified in the cytosol fractions of the human endometrium and the myometrium. The endometrial and the myometrial progesterone binding proteins had sedimentation coefficients of 4.5 S and 4.1 S, respectively. Analysis of the bound steroids revealed that, along with progesterone, small amounts of its metabolites (20alpha hydroxy-4-pregnene-3-one, 5alpha-pregnane-3,20-dione, 5alpha-pregnane-20alpha-ol 3-one) were also bound to the receptor proteins. Among the steroids studied for ligand specificity, 5alpha-pregnane-3,20-dione showed the highest competition for progesterone binding sites. Progestational steroids, like chlormadinone acetate and norgestrel, did not compete for the progesterone receptors. The endometrial and the myometrial progesterone binding receptors were thermolabile and protein in nature. The molecular weight of the endometrial progestrone binding protein was about 60,000-67,000 with a molecular (Stokes) radius of 33 A and the frictional ratio of 1.26. The myometrial progesterone binding protein had a molecular weight of 56,000-58,000 with a molecular (Stokes) radius of 31 A and a frictional ratio of 1.23. The binding of corticosterone to the myometrial cytosol was only 22-34%, whereas with progesterone it was 70-95%. A study of the immunoabsorption of the plasma proteins from the endometrial and the myometrial cytosol suggested the presence of specific progesterone binding receptors in the cytosol that were different to plasma proteins. The association constant of progesterone for the endometrial progesterone receptor was 1.9 x 10(9) M(-1) and for the myometrial progesterone receptor it was 1.4 x 10(9) M(-1), values that are higher than the association constant of progesterone for corticosteroid binding globulin, which is 7 x 10(8) M(-1). The evidence suggested that the human endometrial and the myometrial progesterone binding proteins are different to the corticosteroid-binding globulin. PMID- 19999725 TI - The products of pronase digestion of purified blood group-specific glycoproteins. AB - Purified blood group-specific glycoproteins from ovarian cyst fluids were digested with pronase. The glycoproteins, which had dissimilar amino acid compositions, lost varying amounts of amino acids to leave pronase-resistant cores with very similar compositions. The resistant materials, which were glycoproteins with blood group activities similar to those of the starting materials, contained all of the original carbohydrate. Thus the peptide material removed by pronase action must originate from regions of the macromolecule devoid of sugar. These results confirm previous suggestions that two types of region exist in blood group substances. Threonine, serine and proline accounted for about two thirds of the amino acids present in the pronase-resistant regions. The aromatic and the sulphur-containing amino acids were virtually eliminated by the enzyme treatment. The pronase products of the B, H and Le(a) substances had a near 1:1 ratio of galactosamine to hydroxyamino acids. Similar enzyme resistant materials were obtained when the blood group substances were digested with an insoluble derivative of pronase. On gel filtration the enzyme-resistant materials from different blood group substances had, unlike the parent substances, similar elution profiles and peak elution volumes. Ultracentrifugal examination of one of the pronase-resistant materials indicated that it had a molecular weight of 0.7 x l0(6). Alkaline degradation showed that the enzyme-treated substances had a higher and more uniform degree of O-glycosidic substitution of threonine and serine by carbohydrate chains than did the starting materials. PMID- 19999726 TI - Localization and in vitro specificity of histone acetylation. AB - Histones of isolated calf thymus chromatin incubated in the presence of [14C]acetyl-CoA are radioactively acetylated, at specific residues. Incubation of calf thymus chromatin with [14C]acetate, however, gives no radioactive acetylation. The distribution of the radioactivity among the histone fractions corresponds well to that observed after incubation of calf thymus nuclei with [14C]acetate. The most striking observation is the complete absence of acetylation of histone F1 in both experiments. The specific activity of histones acetylated in vitro is much greater than that of histones acetylated in calf thymus nuclei, most likely reflecting a smaller degree of dilution of the labelled acetate donor. Investigation of the specificity of acetylation of each histone fraction reveals that for each histone fraction, specific peptides are acetylated in vitro which correspond to the histone peptides acetylated in calf thymus nuclei. Histones F2a1 and F2a2, however, also demonstrate additional minor sites of acetylation in the in vitro studies. PMID- 19999727 TI - Cyanogen bromide splitting of human immunoglobulin M. AB - Human Waldenstrdm IgM (kappa) (Dau), its polypeptide chains and its F(c)5 micro and Fab micro fragments were split by cyanogen bromide (CNBr). The fragments formed by CNBr were fractionated by gel filtration, ion exchange chromatography and paper electrophoresis. They were characterized in terms of polyacrylamide disc electrophoresis, peptide maps, amino acid composition, end group determinations and limited primary structure determination. Two CNBr fragments were formed from the kappa chain, consistent with the presence of one methionine residue. Five fragments were isolated from the partially S-sulfonated micro chain. Three additional fragments were released after destruction of all disulfide bonds. The present data are unclear as to whether there are eight or nine CNBr fragments released. The comparison of CNBr pieces from the IgM, mu chain, F(c)5mu and Fab mu affords a tentative arrangement of their order, as well as the relative location of the disulfide bonds within the molecule. PMID- 19999728 TI - Effect of reductive alkylation on catalytic properties of glycogen phosphorylase B. enzyme derivatives with changed nucleotide affinities. AB - Glycogen phosphorylase b modified by NaBH4 and aliphatic aldehydes of varying chain length: ranging from acetaldehyde to n-heptanaldehyde were purified by heat treatment. Kinetic studies showed that the various purified enzyme derivatives were similar to native phosphorylase b in kinetic properties with respect to glucose-I-P. They, however, exhibited different affinities toward AMP. For acetaldehyde, propionaldehyde and butyraldehyde modified phosphorylases b, increase in chain length of the modifying aldehyde resulted in a decrease in AMP affinity of the modified enzyme. For the other aldehydes increase in chain length resulted in an increased AMP affinity of the modified enzyme. Consequently, the apparent values of butyraldehyde and heptanaldehyde modified phosphorylase b exhibited greatest deviation from that of native phosphorylase b. These two enzyme derivatives were studied in more detail. The activation of both enzyme derivatives by AMP could be greatly stimulated by spermine at suboptimal levels of AMP. Both derivatives could be activated by IMP, UMP or CMP in addition to AMP. The extents of activation of heptanaldehyde modified phosphorylase b by these nucleotides were higher than those found for native phosphorylase b. Thus, under certain conditions which included using suboptimal AMP or using IMP, UMP or GMP instead of AMP, heptanaldehyde modified phosphorylase b had much higher catalytic activity than native phosphorylase b. The interactions between burtyraldehyde or heptanaldehyde modified phosphorylases b with IMP were respectively weaker or stronger than that between native phosphorylase b and IMP. The interactions between butyraldehyde or heptanaldehyde modified phosphorylase b with glucose-6-P, an inhibitor of phosphorylase b partially competitive with respective to AMP, were also respectively weaker or stronger than that between the inhibitor and the native enzyme. PMID- 19999729 TI - The selective chloromercuration of insulin. AB - A procedure has been described for the modification of the amino group of glycine methyl ester, and the amino groups of the hormone, insulin, with the reagent p nitrophenyl-p-chloromercuribenzoate, effecting the introduction of the chloromercuri group at various defined sites in the protein molecule. Reaction for 30 min in dimethylformamide afforded several p-chloromercuribenzoyl-insulin derivatives which were characterized by DEAE-Sephadex and Sephadex G-50 chromatography, amino acid analysis, ultraviolet spectroscopy, deamination and trypsin digestion. These derivatives were also shown to possess high biological activity as judged by the glucose oxidation and radioimmunoassays. PMID- 19999730 TI - Chemical modification of the histidine residue located at the active site of ficin. AB - A fully active derivative of ficin was prepared by reversible blocking of its active thiol group with sodium tetrathionate and by oxidation of a methionine residue with sodium metaperiodate. Treatment of this active derivative with bromoacetone in the presence of 2 M urea, pH 6.5, led to its complete inactivation, and the sole change in its composition was the loss of one of its two histidine residues. A comparison of the peptide maps of pepsin digests of the bromoacetone-treated enzyme with its untreated control revealed the disappearance of one of the three histidine-containing peptides present in the control. This particular peptide was shown to have the sequence His-Ala-Val-Ala which is identical to the sequence around the histidine which can be cross-linked to the active thiol of ficin with dibromoacetone (Husain, S. S. and Lowe, G. (1970), Biochem. J. 117, 341). During the course of these studies a very sensitive assay for measuring the activity of ficin was developed which employs p-nitrophenyl-N alpha-carbobenzoxy-L-citrullinate as the substrate. PMID- 19999731 TI - The androgen cytosol receptor in the ventral prostate of the rat. Methodological problems. AB - The phenomena of aggregation and stability of the 8-S androgen receptor in the cytosol fraction from rat ventral prostate have been studied. When glycerol was present both in the cytosol fraction and in the sucrose density gradients, a well defined and easily reproducible 8-S peak was obtained after incubation with tritiated 5alpha-dihydrotestosterone. Without glycerol in the in vitro system the receptor appeared as polydispersed aggregates. Addition of glycerol to the steroid-free cytosol made it possible to store the cytosol at -70 degrees C or 196 degrees C for at least I week without losing 8-S receptor activity. The counting efficiency was studied in different liquid scintillators. A simple toluene scintillation liquid gave the highest counting efficiency. PMID- 19999732 TI - An "extension" of the carbohydrate binding specificity of concanavalin A. AB - Evidence based on the quantitative precipitin method and hapten inhibition technique demonstrates that concanavalin A may interact with internal 2-O-linked alpha-D-mannopyranosyl residues as may occur in glycoproteins and polysaccharides. PMID- 19999733 TI - The precipitation of enzymes from cell extracts of Saccharomyces cerevisiae by polyethyleneglycol. AB - The equilibrium precipitation by polyethyleneglycol of alcohol dehydrogenase, fumarase and invertase from Saccharomyces cerivisiae has been studied. The precipitation can often be represented by a simple linear equation analogous to Cohn's equation for salting-out: log S = X-a x C where S is the protein solubility and C the concentration of polymer, X and a are constants. A more complex form of the equation log S + fS = X-a x C where f is a protein self interaction coefficient, is sometimes necessary, particularly at high protein concentrations and at pH values distant from the isoelectric point. Protein solubility with respect to polyethyleneglycol is influenced by temperature and ionic strength and particularly by pH and protein concentration. Different results were obtained by discrete and sequential addition of polyethyleneglycol. Effective fractional separation of the enzymes is restricted to protein concentrations below about 10 mg/ml. PMID- 19999734 TI - Esterase activity associated with secretory IgA and free secretory component preparations from human milk. AB - Secretory immunoglobulin A (IgA) and free secretory component were purified from human milk using methods involving (NH4)2SO4 precipitation, ion-exchange and gel filtration chromatography. It was found that the preparations, although apparently pure by conventional criteria based on immunoelectrophoresis, zone electrophoresis, or ultracentrifugation, were consistently contaminated by two different esterases. These enzymes could apparently not be removed by further ion exchange chromatography and/or gel filtration. They could, however, be eliminated by passage on an immunoabsorbent made from anti-free secretory component antiserum. PMID- 19999735 TI - The nuclear acidic proteins from haploid and diploid cell states of Physarum polycephalum. AB - The free-living haploid gametes of the Myxomycete Physarum polycephalum are biflagellate phagocytic cells displaying gross morphological and biochemical differences as compared to the diploid plasmodium. The residual acidic chromatin and nucleolar proteins from gametes compatible for mating-type allele have been examined and compared to identical fractions from the diploid plasmodium. The proteins from both gamete mating types are identical yet major differences in the complement of these proteins occur following karyogamy and plasmodial formation. These differences occur in a fraction of acidic chromatin proteins previously shown to contain polypeptides which undergo dramatic changes during differentiation of the diploid plasmodium. PMID- 19999736 TI - A model for the formation and structure of casein micelles from subunits of variable composition. AB - Sedimentation coefficients were determined for association products in systems containing SH-kappa-casein, SH-kappa-casein-alpha(S1) or beta-caseins (1:1, by weight) and SH-kappa-casein-alpha(S1)-casein-beta-casein (2:1:1, by weight) under a variety of environmental conditions. As the concentration of NaCl is increased or, as the concentration of CaCl2 is increased at 0.05 M NaCl, the system containing SH-K-casein alone acts quite differently from those which also contain the Ca2+ -sensitive caseins. It is concluded that electrostatic as well as hydrophobic attractions are important between SH-kappa-casein monomers. Viscosity and sedimentation measurements combined indicate that nearly spherical polymers containing 25-30 monomers each and with radii of approx. 10 nm are formed in all systems. It is considered that these polymers are micelle subunits in which the non-polar portion of each monomer is oriented radially inward while the charged acidic peptides of the Ca2+ -sensitive caseins and the hydrophilic, carbohydrate containing portion of kappa-casein are near the surface. Asymmetric distribution of kappa-casein in a micelle subunit results in hydrophilic and hydrophobic areas on the subunit surface. In this situation, aggregation through hydrophobic interactions forms a porous micelle. Micelle growth is limited by the eventual concentration, at the micelle surface, of subunits rich in kappa-casein. This model successfully accounts for the properties of equilibrium and nonequilibrium micelle systems, for the destabilization of the micelle by dephosphorylation of Ca2+ -sensitive caseins or by high concentrations of NaCl or CaCl2 and for the events associated with clot by rennin action. PMID- 19999737 TI - Inhibition of C1r, C1s and generation of C1s by amidino compounds. AB - A series of amidino compounds has been investigated for their inhibitory effects on C1s, C1r, and generation of C1s. Diamidines consisting of two amidinophenyl residues linked in para position by a molecular bridge proved to be the strongest competitive inhibitors of C1s, whereas those linked in meta position were the strongest competitive inhibitors of C1r. They inhibited the overall generation of C1s when added to the system containing three subunits of C1 and Ca2+. Diphenylamidines were more active than single ring amidines. Of all the compounds tested, dibromopropamidine was the most effective inhibitor of C1s with Ki value 3 x 10(-5) M and deltaF' values 6.4 kcal x mole(-1), whereas amicarbalide and M and B 4596 were the strongest inhibitors of C1r with Ki values 3.5 x 10(-5) M and 3.25 x 10(-5) M and deltaF' values 6.3 and 6.34 kcal x mole(-1), respectively. Epsilon-Aminocaproic acid was also included in this study for comparison purposes and was found to be inert as to its effects on these reactions. The possibility that some of these amidino compounds might prove to be useful for treatment of hereditary angioneurotic edema is discussed. PMID- 19999738 TI - Demonstration of three anomalous plasma proteins induced by a vitamin K antagonist. AB - With the aid of precipitating antibodies against the bovine coagulation factors II, IX, and X three immunologically non-identical proteins can be demonstrated that are induced by the administration of a vitamin K antagonist (phenprocoumon). Each of these proteins is immunologically identical to one of three coagulation factors mentioned. The proteins differ from normal coagulation factors (a) by a lack of biological activity; (b) by a faster anodic migration rate in the presence of Ca2+. The proteins appear in the plasma concomitantly with the decrease of the normal factor. PMID- 19999739 TI - Fluorescent thiol reagents. VI. N-(1-Anilinonaphthyl-4)maleimide; a fluorescent hydrophobic probe directed to thiol groups in protein. AB - N-(1-Anilinonaphthyl-4)maleimide (ANM) (m.p. 207-208.5 degrees C), nonfluorescent, reacts selectively with thiols to give addition products which are strongly fluorescent (excitation maximum 355 nm, emission maximum 448 nm, in ethanol). 0.7 mole of ANM was introduced into thiol groups of egg albumin without modifying any other amino acid residues. In the fluorescence spectra of the reaction products of ANM with thiols, the quantum yields increase and the emission maxima shift towards the blue as the solvent polarity decreases. This remarkable solvent dependence was described in terms ofZ value of the solvents. ANM is thus expected to be a useful fluorescent hydrophobic probe directed to thiol groups in protein. PMID- 19999740 TI - Dielectric hysteresis in proteins. AB - Solid films of ovalbumin and of sodium polyglutamate both show dielectric constants of the order several hundred to several thousand together with marked dielectric hysteresis. It is suggested that these properties may result from ferroelectric behaviour and, if widespread among proteins, must have considerable physiological significance. PMID- 19999742 TI - The centrality of theoretical thinking and the value of empirical evidence. PMID- 19999741 TI - Purification of 125I-labelled lysine-vasopressin by affinity chromatography on sepharose-bound neurophysins. AB - Purification of 125I-labelled lysine-vasopressin has been achieved by affinity chromatography on a Sepharose 4 B conjugate of porcine neurophysins. This affinity absorbent did not retain halogenated hormone, while native lysine vasopressin was bound on the solid support. The specific activity of purified iodinated lysine-vasopressin was 1700-1800 Ci/g, corresponding to one iodine atom per mole. By comparison with an unpurified tracer, a five times increase in the minimum detectable dose was obtained in the vasopressin radioimmunoassay. PMID- 19999743 TI - Complexity in practice environments. PMID- 19999744 TI - Reviews, systematic reviews, overviews: "What's it all about, Cochrane"? PMID- 19999745 TI - Cultural determinants of help seeking: a model for research and practice. AB - Increasing access to and use of health promotion strategies and health care services for diverse cultural groups is a national priority. While theories about the structural determinants of help seeking have received empirical testing, studies about cultural determinants have been primarily descriptive, making theoretical and empirical analysis difficult. This article synthesizes concepts and research by the author and others from diverse disciplines to develop the midrange theoretical model called the Cultural Determinants of Help Seeking (CDHS). The multidimensional construct of culture, which defines the iterative dimensions of ideology, political economy, practice, and the body, is outlined. The notion of cultural models of wellness and illness as cognitive guides for perception, emotion and behavior as well as the synthesized concept of idioms of wellness and distress are introduced. Next, the CDHS theory proposes that sign and symptom perception, the interpretation of their meaning, and the dynamics of the social distribution of resources are all shaped by cultural models. Then the CDHS model is applied to practice using research with Asians. Finally, implications for research and practice are discussed. PMID- 19999746 TI - Self-care demands of persons living with an implantable left-ventricular assist device. AB - Heart failure (HF) is a growing public health problem affecting approximately 23 million people worldwide. Treatment options for advanced HF patients have moved beyond pharmacologic therapy to include left-ventricular assist devices (LVADs). Patients with an LVAD must manage a complex regimen of care. This article proposes the use of self-care deficit nursing theory (SCDNT) as a framework to identify and organize the care needs of patients with an LVAD from a nursing perspective. Within SCDNT, self-care refers to actions designed to meet self-care requisites to achieve regulatory goals. When formalized, self-care requisites have two components: the general action to be taken and a "factor" to be controlled. The reformulation of health-deviation self-care requisites common to LVAD patients are presented and may serve as an exemplar for other technology assisted living situations. The strengths and limitations of using the SCDNT for patients with such complex needs, as well as implications for clinical practice, research, and advancement of nursing science, are examined. PMID- 19999747 TI - A theory of the relational work of nurses. AB - The subject of nurses' relational work articulates a reality that exists but is not seen. However, it is the awareness of this reality that leads to the discovery of where nurses hold their professional power. A theory of the relational work of nurses is the derived theory presented in this article from the parent theory of the relational work of women (Fletcher, Jordan, & Miller, 2000). The new theory exposes the relational work of nurses as the source of their power and value. It explains the relational dynamics of nurses' work, its inherent value, and the need for it to be explicitly recognized as the process by which nurses achieve positive patient and professional outcomes. The new theory proposes that because relational work is institutionally "disappeared", nurses therefore demonstrate lack of caring, patient detachment, and burnout. This theory, if tested, would be an attempt to articulate and make valuable the skills and strengths that nurses enact to get their job done. PMID- 19999748 TI - Therapeutic horticulture in clinical depression: a prospective study. AB - Clinically depressed persons suffer from impaired mood and distortion of cognition. This study assessed changes in depression severity and perceived attentional capacity of clinically depressed adults (N=18) during a 12-week therapeutic horticulture program. The Beck Depression Inventory (BDI) and Attentional Function Index (AFI) were administered at baseline, twice during (4 and 8 weeks), and immediately after the intervention (12 weeks), and at a 3-month follow-up. Experiences of being away and fascination related to the intervention were measured at 4, 8, and 12 weeks. The mean BDI score declined 9.7 points from pretest (27.3) to posttest (p < .001) and were clinically relevant (deltaBDI > or =6) for 72% of the cases. The mean AFI score increased 10.2 points from pretest (68.8) to posttest (p = .06). The greatest change in BDI and AFI scores occurred in the initial weeks of the intervention. The reduction in BDI scores remained significant and clinically relevant at the 3-month follow-up (N=16). The decline in depression severity during the intervention correlated strongly with the degree to which the participants found that it captured their attention. Therapeutic horticulture may decrease depression severity and improve perceived attentional capacity by engaging effortless attention and interrupting rumination. PMID- 19999749 TI - Prevalence of rhinitis in Polish population according to the ECAP (Epidemiology of Allergic Disorders in Poland) study. AB - The prevalence of allergic disorders, especially allergic rhinitis (AR), has dramatically increased in the past few decades and multicentre, standardized, randomized epidemiological studies are required to quantify this phenomenon in Poland. AIM: The aim of the study was to estimate the prevalence of rhinitis and allergic rhinitis in Poland. MATERIAL AND METHOD: The ECAP study was conducted using the ECRHS II and ISAAC questionnaires translated into the Polish language and validated, in selected nine regions of Poland, including eight cities and one rural area. The respondents within the regions were selected by means of multistage proportional stratified random sampling based on the identity number (PESEL) as the operat. The survey was conducted in 20,454 subjects (response rate of 41.9%) and 18,617 questionnaires were valid. Approximately 25% of the subjects (n=4783) were subsequently evaluated by clinicians (response rate of 43.4%). RESULTS: Rhinitis was self-reported by 36.08% of the respondents (37.8% of 6- 7 year-olds, 34.5% of 13- 14-year-olds, and 36.0% of adults). The lowest prevalence rate was in the rural region (22.9%). Allergic rhinitis (AR) was self-reported by 22.54% of the respondents (23.6% of 6- 7-year-olds, 24.6% of 13- 14-year-olds, and 21.0% of adults). Again, the lowest prevalence rate was in the rural region (16.0%). AR was more frequent in males (24.0%) than in females (21.2%) (OR = 1.079; 95% CI: 1.044-1.116). AR was actually diagnosed by a clinician in 28.9%, including intermittent AR in 47.7% and persistent AR in 52.3%. Seasonal AR was diagnosed in 15.55%, and perennial rhinitis in 15.2%. CONCLUSION: Allergic rhinitis is common in Poland as it affects nearly 25% of the population and it is a major social problem. Standards of early detection and prevention of allergic rhinitis should be introduced. PMID- 19999750 TI - [The systems of classification of laryngo-tracheal stenosis]. AB - Laryngo-tracheal stenosis (LTS) is a relatively rare disease, which develops in very heterogenous group of patients. Careful diagnostics and classification are essential for planning the following treatment and assessing its results. To classify and treat LTS, endoscopic and imaging techniques are required. In this publication basing on the literature review different systems of LTS classifications are described in details. Additionally basing on our clinical experience the probe of choosing the best classification for Polish ENT and thoracosurgery departments was undertaken. In our opinion the use of complex grading systems is unreal in everyday work by Polish ENT surgeons and thoracosurgeons. We suggest to implement the Cotton's system and Medical Research Council dyspnea scale as the optimal way to classify and asses the outcomes of LTS treatment. The former system is well known and widely advocated in Poland, the latter requires a modicum of time from the physician. Both scales, as a combination of subjective and objective parameters show a strong correlation and therefore complement each other. PMID- 19999751 TI - [The reasons of laryngo-tracheal stenosis: a review of literature and analysis of 124 patients]. AB - Laryngo-tracheal stenosis (LTS) is a relatively rare disease, caused usually by iatrogenic reasons (intubation, tracheotomy, radiotherapy and surgery of the larynx and trachea) or by trauma. However they may be caused by another rare diseases effecting the wall of the larynx and trachea (primary stenosis) or by compression of pathological process near by (secondary stenosis). Among the primary stenosis except iatrogenic and posttraumatic there are also: congenital, postinfective (tuberculosis, syphilis, scleroma, diphtheria, histoplasmosis and the other), in connective tissue diseases (Wegener's granulomatosis, relapsing polychondritis), in blister diseases (pemphigoid cicarticans, epidermolysis bullosa hereditaria), in amyloidosis, in sarcoidosis and tracheobronchopathia osteochondroplastica. Secondary stenosis may be effected by thyroid diseases, diseases of the big blood vessels of the thorax, hypertrophic thymus, enlargement of the lymph nodes near by the trachea, tumors and cysts of the neck and mediastinium, retrotracheal abscess and cold abscess in vertebral column tuberculosis. If the reason of stenosis is unknown, the stenosis is called idiopathic. In this publication basing on the literature review the reasons of LTS were described in details. We also present our clinical experience concerning 124 patients. The authors underline, that in most of patients the reason of LTS can be find after taking the history of a patient. However, in the cases, when the reason is not clear, a lot of specific examination should be done to find another rare reason, and only after exclusion it the diagnosis of idiopathic LTS can be established. PMID- 19999752 TI - [Endoscopic management of cerebrospinal fluid rhinorhea]. AB - Endoscopic surgery due to its high efficacy and relatively minor intraoperative trauma became a standard of transnasal csf leak treatment. THE AIM OF THIS STUDY: was to present, basing on our own experience, diagnostic and therapeutic methods of treatment of various types of csf rhinorhea. MATERIAL AND METHOD: All together 14 endoscopic skull base reconstructions were performed. The bony defect and dural perforations were closed using: fat grafts, septal bone or cartilage, mucosal flaps of the septum or nasal concha, artificial dura and collagen sponge with fibrin glue. The defect was localized by high resolution computed tomography, magnetic resonance cisternography, neuronavigation system and fluorecein test. RESULTS: Complete closure of the defect and resolution of the csf leak after the first attempt was achieved in 92,3% of patients. In one case the rhinorhea was successfully stopped after the second reconstructive procedure. No adverse reaction of intrathecal fluorescein administration were noted in our patients. CONCLUSIONS: Endoscopic surgeons have a wide variety of diagnostic tools and operative techniques at their disposal for precise localization and management of csf rhinorhea. The key for high efficacy of the treatment is appropriate selection of diagnostic and surgical method depending on cause, type, size and localization of the leak. PMID- 19999754 TI - [The metastases to the head and neck region from distant primary localization]. AB - INTRODUCTION: Distant metastases to the head and neck region, with primary tumor located outside the head and neck, presents often a diagnostic problems, especially when metastases are an exclusive signs and symptoms of the disease. In most of this cases there is no chance for curative therapy, and optimum palliation is the goal of the treatment. AIM: The description and analysis of clinical and prognostic factors, which could establish the optimum diagnostic formula, and predict the prognosis in distant metastases to the head and neck region. MATERIAL AND METHODS: The study of 44 cases diagnoses and treated in 2001 2006 in Dept. ORL H&N Surgery. The analysis was based on current observations and case records documentation. RESULTS/CONCLUSIONS: In the most of cases (39/44) the distant metastases to the head and neck regions presented a part of generalized malignant neoplastic disease, but in 14/44 cases that was a first warning symptom. All of this 14/44 cases presented as a metastases to the IV and V lymph nodes region. The most common primary sides were found in the lung. The average survival within the analyzed group was only 7,2 months. PMID- 19999753 TI - [Dendritic cells in hypertrophied adenoid at children with otitis media with effusion]. AB - INTRODUCTION: The adenoids are organized as lymphoepithelial structures that play an important role in protecting both the upper respiratory and alimentary tract regions. This functions requires dendritic cells (DC) which are one of the major populations of immune cells. Due to the presence of specific receptors (DC) are able to respond to both intra- and extracellular antigens. Dendritic cells activating immunological response in tonsil contribute formation immunologic competent cells on necessity of rolling inflammatory process in middle ear. AIM OF STUDY: An investigation was executed in hypertrophied adenoids with or without otitis media with effusion. METHODS: By flow cytometry percentage of CD11c+ myeloid DC and 123+ plasmacytoid DC in hypertrophied adenoid and hypertrophied adenoid and otitis media with effusion was analyzed. RESULTS: The percentage of CD11c+ myeloid DC and 123+ plasmacytoid DC was similar in hypertrophied adenoid and otitis media with effusion compored to the control group. CONCLUSIONS: Our data show that part of dendritic cells has not on course of inflammatory process influence rolling in middle ear. PMID- 19999755 TI - [Problem of infections at ENT Department of Poznan University of Medical Sciences based on the material from 2006-2007 years]. AB - INTRODUCTION: Infections at ENT wards constitute an essential clinical, organizational and economic issue. AIM: The purpose of this work was to present the problem of infections at ENT Clinic of Poznan University of Medical Sciences based on the material from 2006-2007 years. MATERIAL AND METHODS: In 2006-2007 in The Department of Otolaryngology in Poznan there were treated 7052 patients. During that time 476 biological samples were taken to carry out a bacteriological examination. They came from: 74 postoperative wounds, 43 tracheotomy areas, 29 tracheoesophageal fistulas, 9 salivary fistulas, 105 ears, 71 noses, pharynxes and oral cavities, 44 body fluids, 18 paranasal sinuses, 11 central venous catheter areas, 23 urine samples, some constituted BAL, sputum, cyst content, materials from decubitus ulcer and gastrostomies areas. 21 materials were taken from abscesses and phlegmons, 32 from patients with purulent complications of chronic otitis media, 18 from patients with complications of chronic sinusitis and in 1 case there was taken in hospital conditions peritonsillar abscess content. RESULTS: 119 biological samples were aseptic, in 43 samples there were no pathological cultures, in 314 samples pathological bacterial flora was found. The most often isolated pathogens constituted: in postoperative wounds Staphylococcus epidermidis, in tracheotomy areas-Enterococcus faecalis, in tracheoesophagal fistulas-Streptococcus viridans, in ears-Pseudomonas aeruginosa, in materials from noses, pharynxes and oral cavities-Staphylococcus aureus, while in materials from abscesses, paranasal sinuses and central venous catheter areas Staphylococcus epidermidis. CONCLUSIONS: All cases were analyzed in detail. There was made a correlation between isolated pathogens and: a kind of pathology, surgical procedure, general patient condition, length of hospitalization. PMID- 19999756 TI - [The value of the Halmayi-Curthoysa test in neurological diagnostic procedures]. AB - THE AIM OF THE STUDY: The goal of the study was to state the usefulness of the Halmagyi-Curthoys' test in the patients with unilateral vestibular weakness. The influence of the heaviness of the canal paresis, spontaneous nystagmus and compensation on the test results was observed. MATERIAL AND METHODS: The investigation was performed in 68 patients with unilateral vestibular weakness of the various origin. Halmagyi-Curthoys' test was done using passive way. RESULTS: In healthy subjects the test results were negative. In patients: 47% demonstrated positive test, 44,1% negative proof, in 8,9% test was difficult to estimate. CONCLUSIONS: The obtained results pointed out that Halmagyi-Curthoys test is useful as orientation, preliminary examination, generally in cases with deep canal paresis (according to Dix- Hallpike's classification). PMID- 19999758 TI - [The usefulness of some vestibular tests in the diagnosis and treatment of patients with vertebro-basilar insufficiency]. PMID- 19999757 TI - [Utility of PET/CT in evaluation of haed and neck carcinoma--preliminary report on the basis of presented cases]. AB - 18F-fluorodeoxyglucose (FDG)-positron emission tomography (PET)/computed tomography (CT) offers advantages over PET alone, which is limited by poor anatomic localization and CT alone, which provides morphologic data only. Fusion of PET and CT images allows for precise target delineation for head and neck cancer compared with CT alone. The aim of this article is to evaluate the effect of the use of PET/CT in patients with head and neck cancer. We present four patients whose management was influenced by the implementation of combined PET/CT imaging. We conclude that PET/CT allows detection of recurrent neoplasm, carcinoma of unknown origin, second primary sites, distant metastases, tumor surveillance, staging, and planning radiation therapy. PMID- 19999759 TI - [Atopy and other factors influence on frequency appearing of different cytological types of nasal polyps]. PMID- 19999760 TI - A review of biomechanic and aerodynamic considerations of the avian thoracic limb. AB - The wings are the most important part of the flight apparatus of a bird and consist of feathers, bones, muscles, nerves, and patagial skin flaps. The complex kinematics of wing beats and the perfect control of aerodynamics make avian flight possible. An impaired flight can be viewed as an avian lameness; therefore, a better understanding of avian locomotion can help to diagnose and to evaluate the avian patient, especially when perfect flight is required for release of wild birds. Every condition affecting a specific part of the wing can lead to serious biomechanic and aerodynamic consequences during flight. This review summarizes wing mechanics that might be of clinical relevance for avian practitioners considering the current experimental and theoretical scientific knowledge available on avian flight in conjunction with observations of birds in various wildlife centers. PMID- 19999761 TI - Comparison of treatment protocols for removing metallic foreign objects from the ventriculus of budgerigars (Melopsittacus undulatus). AB - To compare the efficacy of treatment protocols recommended to aid passage of metallic foreign objects from the ventriculus of birds, a 1-mm metal sphere, made from solder wire, was placed into the crop of each of 44 budgerigars (Melopsittacus undulatus). After survey radiographs confirmed the spheres were lodged in the ventriculus, birds were divided into 6 groups. Each group received 1 of 6 different treatment protocols: psyllium with grit, acidic drinking water, fine grit, coarse grit, cathartic emollients (peanut butter and mineral oil), and a control group. All birds were treated simultaneously with a chelating agent, dimercaptosuccinic acid (DMSA), to prevent heavy-metal toxicosis. Successive survey radiographs were used to monitor elimination of the spheres from the digestive tract. Of all protocols tested, birds treated with either fine or large grit had the shortest mean elimination time of the metal spheres. These results indicate that administration of grit particles, either fine or coarse, appears to be effective in hastening the passage of metallic foreign objects from the ventriculus of budgerigars. PMID- 19999762 TI - Laboratory reporting accuracy of polymerase chain reaction testing for psittacine beak and feather disease virus. AB - Diagnostic assays that use polymerase chain reaction (PCR) methods are increasingly available to veterinarians. Psittacine beak and feather disease virus (family Circoviridae, genus Circovirus) is a pathogen of clinical importance for which PCR assays have been developed. Several laboratories offer this diagnostic assay in the United States; however, there is little information on the sensitivity, specificity, or accuracy of these assays. In this study, known positive (n = 10) and negative (n = 10) samples were sent to 5 commercial laboratories. Accuracy was 100% for 2 laboratories, 95% for 2 laboratories, and 71% for 1 laboratory; the accuracy of the latter laboratory was affected because of a specificity of 20%. These results suggest that although the results from most laboratories are highly accurate, both false-positive and false-negative results are occasionally reported by at least 2 laboratories. These results also suggest that at least 1 laboratory may be generating large numbers of false positive results. PMID- 19999763 TI - Effects of midazolam on ketamine-xylazine anesthesia in guinea fowl (Numida meleagris galeata). AB - To determine the effect of midazolam on ketamine-xylazine anesthesia, 20 guinea fowl (Numida meleagris galeata) were randomly divided into 2 groups. Birds in group KX were anesthetized with ketamine (15 mg/kg i.m.) and xylazine (2.5 mg/kg i.m.), whereas the birds anesthetized in group KXM received midazolam (0.3 mg/kg i.m.) in addition to the ketamine and xylazine protocol. The onset of anesthesia, duration of analgesia, duration of recumbency, and recovery time were determined. Heart and respiratory rates as well as cloacal temperatures were recorded immediately after drug administration and at 10-minute intervals until the birds were sternally recumbent. Analgesia was assessed as a response to artery forceps applied to the digit and skin proximal to the tarsal joint. The mean (SD) duration of analgesia in the group KXM birds was 37.4 +/- 23.5 minutes, whereas no analgesia was apparent with the group KX birds. The duration of recumbency was significantly longer and respiratory rates were significantly lower in group KXM birds compared with those in group KX. Adverse effects were minimal and included diarrhea (n = 1) and hypersalivation (n = 2) in group KX birds, and regurgitation (n = 2) in group KXM birds. Midazolam administered intramuscularly appeared to improve the anesthetic quality of ketamine and xylazine in guinea fowls without adversely affecting safety. PMID- 19999764 TI - Avian paramyxovirus serotype 3 infection in Neopsephotus, Cyanoramphus, and Neophema species. AB - An outbreak of avian paramyxovirus serotype 3 (APMV-3) infection occurred in a private bird collection consisting mainly of Neophema species. Two Bourke's parrots (Neopsephotus bourkii) and one red-crowned parakeet (Cyanoramphus novaezelandiae) were examined after developing torticollis and circling. The 3 birds were euthanatized and submitted for necropsy. Brain and liver samples from the 3 birds were homogenized and inoculated into the allantoic cavity of 10-day old embryonated specific-pathogen-free chicken eggs. An APMV-3 isolate replicated but did not induce embryonic mortality after up to 5 consecutive passages. Allantoic fluid from the second passage was used in hemagglutination testing performed on chicken erythrocytes, which resulted in a titer of 64 hemagglutinating units. Avian paramyxovirus serotype 3 was identified in the allantoic fluid, and a reverse transcription polymerase chain reaction verified the isolate as APMV-3/parakeet. An eventual mortality rate of 70% occurred in the Neophema species, but a group of budgerigars (Melopsittacus undulatus) and parrotlets (Forpus species) kept in the same facility showed no clinical signs during the outbreak. PMID- 19999765 TI - A multimodal approach to management of suspected neuropathic pain in a prairie falcon (Falco mexicanus). AB - An adult male prairie falcon (Falco mexicanus) was presented for evaluation and treatment of self-inflicted wounds along the right proximal patagium. The bird had started self-traumatizing approximately 1 month after fracturing the right metacarpus, although the fracture had stabilized, surface wounds had healed completely, and treatment with a nonsteroidal antiinflammatory drug had been administered. The bird was treated with gabapentin (11 mg/kg p.o. q12h), ketamine (0.5 mg/kg i.m. q24h), and low level laser therapy (LLLT) (<5 mW, 630-680 nm, 5 second application per site) for 3 weeks, but, despite initial improvement, new self-inflicted wounds appeared at the same site. Approximately 1.5 months later, a radial and medianoulnar nerve block was performed by using bupivacaine (2 mg/kg) and medetomidine (0.5 microg/kg). In addition, the dosing interval of ketamine was increased to q12h, and the dose of gabapentin was increased 7.5 fold. A higher energy and wavelength of LLLT (1040 mW, 830 nm, 2 J/cm2) was applied once to the injured region and fracture site, then the original LLLT protocol was applied once daily. After 2.5 months, the wounds healed completely and no further mutilation took place. Once deemed ready for release, the falcon was returned to the wild after 181 days in captivity. This is the first reported application of successful multimodal analgesia in a raptor with uncontrolled neuropathic pain. PMID- 19999766 TI - Two cases of neoplasia of basal cell origin affecting the axillary region in anseriform species. AB - Neoplasms of the skin are occasionally seen in domestic birds but are uncommon in nondomestic birds. An 8-year-old male hooded merganser (Lophodytes cucullatus) was presented with bilateral axillary ulcerative lesions that improved but did not resolve with empiric antibiotic and antifungal therapy. Skin biopsies were taken, and bilateral feather folliculomas were diagnosed on histopathologic examination. The duck was euthanatized because of the poor prognosis. A 9-year old Indian runner duck (Anas platyrhynchos) was presented with an ulcerative lesion, with pseudomembrane and serocellular crust affecting the axillary region. This mass was diagnosed as a basosquamous carcinoma. The mass was surgically excised, and no recurrence was observed. Feather folliculomas are usually considered benign neoplasms in domestic birds and may be primarily ulcerative, exudative, bilateral, and symmetric in location. Basosquamous carcinoma may have a similar gross appearance. It is unknown if the axillary region may be an area with increased incidence of neoplasia in birds. This appears to be the first report of feather folliculoma and basosquamous carcinoma in Anseriforme species. Feather folliculomas and other neoplasms, such as basosquamous carcinoma, should be considered as a differential diagnosis in ulcerative or proliferative skin lesions in birds. PMID- 19999767 TI - Veterinary marketing--what works and what doesn't. PMID- 19999768 TI - What is your diagnosis? Mycobacterium avium subsp avium. PMID- 19999769 TI - [Meeting foreigners with curiosity]. PMID- 19999770 TI - [From infection control to family health care: Brazil: development of nursing science]. PMID- 19999771 TI - [Advent and Christmas in nursing homes for the elderly: tolerance in the face of different customs]. PMID- 19999772 TI - [Culture sensitive nursing and health care and nursing facilities: risking curiosity, overcoming strangeness]. PMID- 19999773 TI - [How a Sinti family envisions death with dignity: challenge to transcultural communication]. PMID- 19999774 TI - [Challenging behavior and biography work in the nursing home: recognizing commonalities]. PMID- 19999775 TI - [Medicinal plants for wound care: an alternative method]. PMID- 19999776 TI - [Strike: refusal to perform contracted work: occupational dispute resource of employees]. PMID- 19999777 TI - [Expertise in questions and answers--the nursing duty schedule regulation: vacation during nursing duty time]. PMID- 19999778 TI - [Report of experiences of the University Clinic in the Bern Island Hospital: DRG in routine surgery--patient safety and quality assurance]. PMID- 19999779 TI - [Taking a position in the financial and economic crisis: making the best of it]. PMID- 19999780 TI - [Developement of a standardised questionnaire. Specific interventions for people with dementia in long-term care]. AB - Caring for residents in long-term care who suffer from dementia is a special challenge. In the context of a research project a questionnaire for a mail survey in nursing homes was developed in order to describe specific nursing interventions for dementia in daily practice. The Questionnaire development was done according to acknowledged standards for questionnaire development, using means of operationalisation and external expert rating. Then the questionnaire was tested in a two-phase-pretest. Seven nursing homes were involved in a cognitive pretest. The standard-pretest was sent to 200 nursing homes (response rate 17.5 percent). Evaluation of the questionnaire drafts by means of two-phase pretesting proved to be an effective measure for continuous improvement. The resulting questionnaire should be used to collect data about specific interventions for people suffering from dementia in residential long-term care. PMID- 19999781 TI - [New educational offer for graduate nursing faculty: curriculum offer closes gap in nursing education ]. PMID- 19999782 TI - [Planning a practical instruction situation--4: Leading a nursing specialty discussion in the geriatric department]. PMID- 19999783 TI - [From case to case: respecting the cultural and religious needs of patients]. PMID- 19999784 TI - [Recognizing and understanding illnesses: jaundice as the chief symptom]. PMID- 19999785 TI - Tobacco, alcohol and doctors. PMID- 19999786 TI - The association of iron status with educational performance and intelligence among adolescents. AB - INTRODUCTION: The aim was to identify the association of iron status with educational performance and intelligence of adolescents. METHOD: This was a cross sectional comparative study among adolescents aged 13-15 years. Each iron deficient student was matched with an iron sufficient student from the same school, class and sex. Iron status was based on haemoglobin and serum ferritin levels. The marks for mathematics, science, Sinhala language and social science were considered to assess educational performance. Intelligence was measured by Raven's Standard progressive matrices. All the possible confounders and effect modifiers were considered. Home visits to a sub-sample checked the quality of data. RESULTS: The final analysis included 188 students (94 matched pairs). Neither educational performance nor intelligence showed significant associations with the iron status. The severity of the iron deficiency did not relate to these cognitive variables either. Twenty-three and 8 co-variables showed statistically significant associations with educational performance and intelligence respectively. Following a multiple regression analysis intelligence, the enthusiasm of the student towards learning, occupational ambition, household possession, problems at home and private tuition for mathematics were key factors predicting educational performance. Stunting and educational level of the mother were important factors influencing intelligence. CONCLUSION: Iron status does not play a major role in educational performance and intelligence of school going adolescents. Several factors affect educational performance and intelligence. This study highlights the difficulty in extrapolating the findings of similar studies to different ecological settings. PMID- 19999787 TI - Juvenile victimisation in a group of young Sri Lankan adults. AB - OBJECTIVE: To study the prevalence of juvenile victimisation in a group of young adults. METHOD: A juvenile victimisation questionnaire was distributed among 1322 Sri Lankan undergraduates. The questionnaire consisted of different modules (child maltreatment, conventional crime, peer-sibling victimisation, indirect victimisation, introduction to substances and parental deprivation). RESULTS: The response rate was 90%. The mean age of the cohort was 21.8 years. 59% were females. 44% and 36% had experienced sexual and physical maltreatment respectively. In both categories males were affected more than females (p < 0.001). Physical abuse had commonly taken place at school (51%) and home (40%). Witnessing violence at home was the highest form of indirect victimisation (66%). 10% were introduced to substances in childhood. Usage of substances (cigarettes, alcohol and drugs) was significantly higher in children whose fathers used substances compared to children whose fathers did not (p < 0.001). CONCLUSION: Many children in Sri Lanka are exposed to victimisation. They seem to suffer these in the very environments that should be nurturing and protecting them. PMID- 19999788 TI - STR polymorphisms in Sri Lanka: evaluation of forensic utility in identification of individuals and parentage testing. AB - OBJECTIVES: This preliminary study was carried out to determine the allele frequencies and forensic efficiency parameters for the short tandem repeat loci CSF1PO, TPOX, THO1, D16S539, D7S820, D13S317, vWA, FESFPS and F13B in a test sample population of Sri Lankans. DESIGN: Test samples were obtained from 305 non related individuals originating from all 9 provinces of Sri Lanka. DNA was extracted from whole blood using chelex-100 and amplified by PCR using the GenePrint STR kit and silver stained. Final DNA profiles were analysed for forensic efficiency parameters and paternity indices using PowerStats version 12. Possible divergence from Hardy-Weinberg Equilibrium was tested using the chi square test and exact test. RESULTS: All common alleles in the allelic ladders were found in the test sample studied. PIC values >0.5 for all 9 STR loci indicate this STR system to be informative and useful for identification purposes. The D13S317, vWA and D7S820 loci were found to be the most polymorphic markers of the system studied. CONCLUSIONS: No deviations from Hardy-Weinberg Equilibrium were found for any of the loci examined. The results indicate that the 9 STR loci system described here is suitable for estimating DNA profile frequencies in human identification and forensic and parentage testing for legal purposes among Sri Lankans. PMID- 19999789 TI - Genetics of pre-eclampsia and the weight of babies at birth--clinical and genetic studies in Sri Lanka. PMID- 19999790 TI - Estimation of body surface area: nomogram vs. mathematical equations. PMID- 19999791 TI - Exchange transfusion for intrahepatic cholestasis due to sickle beta thalassaemia. PMID- 19999792 TI - Uncommon presentation of hypereosinophilic syndrome. PMID- 19999793 TI - Gossypiboma. PMID- 19999794 TI - An autologus lower limb prosthesis. PMID- 19999795 TI - Acute diffuse proliferative glomerulonephritis with atypical presentation in Sri Lankan adults--clinical features and outcome. PMID- 19999796 TI - [Side effects of caffeine]. AB - Caffeine is one of the most commonly ingested alkaloids worldwide. It is present in coffee, tea, soft and energy drinks, chocolate, etc. Currently published data has been stressed that the metyloxantine consumption increases the risk of coronary heart disease, arterial hypertension, arterial stiffness, and an elevation of cholesterol and homocysteine plasma concentration. The acute high consumption may also modulate insulin sensitivity and glucose blood level. However, the long-term consumption reduces the incidence of the type 2 diabetes mellitus. When administered in high doses the substance may cause various side effects, related to abnormal stimulation of the central nervous system, decrease tonus of the lower esophageal sphincter, as well as increase risk of miscarriage and intrauterine growth retardation. The final manifestation of side reactions is dependent on the genotype, especially polymorphisms of genes associated with caffeine metabolism, i.e., cytochrome P450-CYP1A2 and catechol-O methyltransferase (COMT). PMID- 19999797 TI - [Effectiveness of heart tumor therapy in the cardiology department during 7 year follow-up]. AB - Neoplasms of the heart are rare. Usually asymptomatic on the early stage are diagnosed incidentally. Among primary heart neoplasms the most often benign tumors are diagnosed--mostly myxomas, whereas the majority of malignant heart tumors are sarcomas. The aim of this paper was to present heart tumors diagnosed in the cardiology department, their symptoms, used diagnostic tests and therapy and to show after therapy quality of life changes. MATERIAL AND METHODS: There were 18 patients included to the study, whom during hospitalization in the cardiology department heart tumors were diagnosed. There were 11 women and 7 men, aged from 33- to 76-years-old (mean 60,5 years). To all of the patients medical interview, physical examination, EKG, UCG and laboratory test were performed. Additionally in some cases computed tomography or magnetic resonance imaging of the chest and coronary angiograms were done. Based on the diagnostic tests results the patients were qualified to conservative or surgical treatment. RESULTS: Among 18 heart tumor patients in 12 cases primary benign tumors were diagnosed (66,6%), 1 patient had primary malignant tumor (5,5%), there were 3 cases of metastatic tumors (16,6%) and 2 patients with non-neoplasmic tumors- clots (11,1%). From 18 subjects with heart tumor 3 patients died because of advanced stage of neoplasmic disease and presence of metastatic tumors in the heart. CONCLUSIONS: Results of the study show, that heart tumors, regardless of development of diagnostic tests, are still diagnosed too late. The study group follow-up proved, that early diagnosis and proper heart tumor treatment prevented complications and improved the quality of life. It is worth to emphasize, that coronary angiogram in some cases allowed to diagnose coronary artery disease, to treat heart tumor and to perform coronary artery by-pass grafting simultaneously. PMID- 19999798 TI - [Usefulness of sialic acid determination to differentiation between alcohol abuse and alcoholic liver cirrhosis]. AB - Sialic acid is a new marker of alcohol abuse. The synthesis and the catabolism of sialic acid takes place in the liver, therefore the status of liver can influence the total serum level of sialic acid (TSA). The aim of this study was to evaluate the effect of alcoholic liver disease on the serum level of TSA and the usefulness of TSA determination to differentiation between alcoholics without liver disease and those with alcoholic liver cirrhosis. MATERIAL AND METHODS: Serum TSA was determined in 63 alcohol-dependent males (aged 23-77 years, average 46 yrs), 46 patients suffering from alcoholic liver cirrhosis (aged 26-66 years, average 48 yrs) and 50 healthy men by enzymatic-colorimetric method. The diagnosis of dependency was made on the basis of ICD-10 criteria, and alcoholic liver cirrhosis was recognized on the basis of physical examination, liver ultrasound, endoscopy and laboratory tests. RESULTS: The TSA concentrations in the alcohol abuse patients were significantly higher than in healthy controls but in alcoholics with liver cirrhosis did not differ in comparison to the controls. The diagnostic power of TSA determination in alcoholic patients without liver disease is high but in alcoholics with liver cirrhosis--low. The elevated level of TSA in alcohol abuse patients distinguish (ROC analysis) alcoholics without the liver disease and alcoholics with the liver cirrhosis. CONCLUSION: Diagnostic power of total sialic acid concentration in alcoholics with healthy liver is higher than in patients with alcoholic liver cirrhosis. Therefore, TSA determination is usefulness diagnostic test for the differentiation between alcoholics without alcoholic liver disease and those with the alcoholic liver cirrhosis. PMID- 19999799 TI - [Laparoscopic cholecystectomy with single incision laparoscopic surgery]. AB - For the last seventeen years laparoscopic cholecystectomy has remained the gold standard procedure for gallbladder surgery The single incision laparoscopic surgery (SILS) approach is a step toward less invasive surgery and the intraumbilical access improves the cosmetic effect (no scar surgery). The aim of the study was to present our preliminary experience in laparoscopic cholecystectomy through single intraumbilical incision. MATERIAL AND METHODS: Three patients with gallbladder symptoms (2 female, 1 male) underwent surgery through single intraumbilical incision. In two cases the procedure was performed with three 5 mm Dexide port approach and in one, with a flexible rubber/gel SILSTM-Port (Covidien, USA) access. A simple 5 mm optics was employed. In two procedures the following disposable instruments were used: roticulator Endo dissect, roticulator Endo grasp, mini Sears scissors as well as standard hook and clips while for the third procedure, a conventional multi-use grasp and roticular Endo dissect. The gallbladder was elevated to the integuments by two sutures to expose Calot's triangle and bladder site. Following exposure, the bile duct and the vesical artery were clipsed and cut routinely to remove the gallbladder in a plastic bag through the bell-button. After procedures no drains were left in the peritoneal cavities. RESULTS: In all three cases the laparoscopic procedure was performed. The median operative time was 88 min. In one female patient, bleeding into the peritoneal cavity was reported and called for laparotomy within a few hours of laparoscopic procedure. Intraoperatively, the branch of vesical artery was sutured to arrest bleeding. The median hospital stay following procedure was 3, 8 and 5 days respectively. After the procedure, the last female patient was observed to have pharyngitis and enlarged jugular lymph nodes that abated after antibiotic therapy. CONCLUSIONS: Single-incision laparoscopic cholecystectomy should be performed by experienced laparoscopic surgeons. For adequate evaluation of the efficacy and safety of this procedure it is necessary to perform further clinical studies on a larger number of patients. PMID- 19999800 TI - [Strategy for secure hemostasis in hemophilia patients undergoing surgery for malignant neoplasms]. AB - Surgery for malignant neoplasms in hemophilia patients is no different from standard procedures in the general population on condition that the normal hemostatic activity of deficient factors: VIII in hemophilia A and IX in hemophilia B in perioperative period is ensured. The aim of the study was to determine the type and frequency of malignant neoplasms in hemophilia patients as well as to provide a strategy for establishing safe hemostatic activity in surgically treated hemophilia patients. MATERIAL AND METHODS: In the period 2003 2008, surgical procedures were performed on 19 hemophilia A and B patients with diagnosed malignant neoplasms of various location. The following cases were diagnosed: 9 colorectal neoplasms, 2 pancreatic carcinomas, 2 larynx carcinomas, 1 stomach carcinoma, 1 liver, 1 nasopharyngeal, 1 testicle, 1 prostate and 1 skin. Seventeen patients were hemophilia A (6 severe, 4 severe with inhibitor, including 2 high titre and 2 low titre, 1 moderate and 6 mild) and 2 were hemophilia B (1 severe, 1 moderate). Patients mean age was 55.8 years (22-82). In factor-replacement therapy for patients with no inhibitor the strategy was to maintain the activity of the deficient clotting factor VIII before the operation at 80-100% of normal value, within the 80-100% range on the 1-3 day following surgery, at 60-80% on days 4-6, at 30-60% on days 7-10 day and at 20-40% on all subsequent days until the surgical wound healed. In hemophilia B patients the levels were about 20% lower. Deficient factor was injected every 8 or 12 hours or administered in continuous intravenous infusion. In hemophilia A patients with high titre inhibitors to factor VIII (above 5 Bethesda units/ml) activated prothrombin complex concentrates (aPCC)-FEIBA (Baxter AG) were used at 50-100 U/kg b.m., every 8-12 hours. RESULTS: The nineteen patients with diagnosed malignant neoplasms (in the period 2003-2008) constituted 0.77% of the overall 2475 hemophilia patients entered into the National Registry Congenital of Hemorrhagic Diathesis. An overall number of 26 surgical procedures were performed in this group of 19 hemophilia patients including 20 procedures for malignant neoplasms and 6 for postoperative complications. All patients survived surgery. Two patients with pancreatic carcinoma died in the postoperative period due to multi-organ failure. Complications occured in 7 (37%) patients including 6 (32%) with bleeding complications: haemopneumothorax (n=1), intraperitoneal bleeding (n=2), abdominal parietal hematoma (n=1), hematuria (n=1), bleeding from esophago pharyngo-cutaneous fistula following total laryngectomy (n=1). Other complications included: ileus (n=1), leakage of pancreato-jejunal anastomosis (n=1), dehiscence of abdominal wound (n=1) and bleeding from duodenal ulcer (n=1). These complications were successfully treated with surgery or endoscopy. CONCLUSIONS: Surgery of malignant neoplasms in hemophilia patients is burdened with a high risk of complications which include bleedings despite adequate replacement therapy and administration of factor eight by-passing concentrates in patients with high titre inhibitor. Therefore surgical procedures involving these patients should be performed in specialized centers with experienced team (surgeon, anaesthesiologist, haematologist) and supported by a laboratory for coagulation disorders. PMID- 19999801 TI - [Pharmacotherapy of patients with mental retardation during psychiatric hospitalization]. AB - Mentally retardated patients are considered to be the worst treated and medical cared group of psychiatric patients. Mentally impaired patients are diagnosed following the same diagnostic criteria as patients with normal intellectual abilities. The diagnosis stated should determine adequate pharmacological treatment schemes, the same as for general population. The aim of paper was to analyse pharmacological treatment administered to mentally retarded patients during psychiatric hospitalization. MATERIAL AND METHODS: The medical documentation of patients hospitalized in Babinski's Hospital in Lodz in 2006 with diagnosis of mental retardation (F70-F79 according to ICD-10). The patients hospitalized throughout the year 2006 with co-morbid psychiatric diagnosis as well as those with only stated mental retardation were taken into account. The analysis was based on questionnaire specially prepared for that paper. The results were statistically analyzed. RESULTS: The most frequent administered treatment among mentally retarded patients during psychiatric hospitalization were antipsychotics of the first generation and mood stabilizers independently on co-morbid psychiatric disorders. CONCLUSIONS: Regardless the level of mental retardation the most rarely administered group of medicines seemed to be antidepressive drugs (statistical importance) and atypical antipsychotics. Mood stabilizers were statistically more frequently administered in patients with severe level of mental retardations. PMID- 19999802 TI - [Ascites and extreme lipid abnormalities as initial symptoms of cystic fibrosis in a 5-years-old girl--case report]. AB - Cystic fibrosis is the most common, multiorgan inherited autosomal recessive disorder. The gene associated with this disease encodes the CF transmembrane conductance regulator (CFTR). The aim of this article is a presentation of 5 years-old girl with cystic fibrosis and associated liver disease as the only manifestation of CF. CASE REPORT: We described the case of a 5.5-years-old girl with ascites and peripheral edema without of the symptoms. Laboratory tests revealed hepatic cell damage with cholestasis, extreme lipid abnormalities and hypocholesterolemia. Based on positive sweat test (CI: 122 mEq/l) and genetic analysis (homozygote deltaF508) the diagnosis of cystic fibrosis was confirmed. Patient was successfully treated with nutritional treatment and pancreatic enzymes. PMID- 19999803 TI - [Thrombus or tumour of the mitral valve--a case of 75-year-old woman with acute coronary syndrome and aortic insufficiency]. AB - Papillary fibroelastomas are rare, primary, benign cardiac tumors most frequently located in the heart valves. They are a potential cause of systemic embolization and may be associated with myocardial ischaemia, transient ischaemic attacks, strokes, or sudden death. We describe the case of 75-year-old woman with diagnosed aortic insufficiency who was admitted to hospital with acute coronary syndrome. The fibroelastoma of the anterior mitral leaflet was diagnosed using transthoracic and transoesophageal echocardiography. We provide a differential diagnosis and review of the literature concerning the pathologic intracardiac masses affecting cardiac valves. PMID- 19999804 TI - [Severe depressive episode with psychotic symptoms on the basis of hypothyroidism]. AB - The prevalence of depressive disorders in hypothyroidism is the subject of long term studies and controversies. The studies on big populations appear to confirm a substantially higher prevalence rate of depression in patients diagnosed with hypothyroidism. Considering interdependence between depression and hypothyroidism the possibility of coincidence of the two disorders cannot be forgotten. The aim of the work is a case of late diagnosed hypothyroidism in the course of which a severe depressive episode with psychotic symptoms occurred. A female patient, aged 39, commenced outpatient psychiatric treatment because of sleeplessness in May 2008. In the following months a lack of appetite, skin dryness and hair loss occurred. As a result of hypothyroidism diagnose she started endocrinological treatment in the Center of Thyroid Illness Treatment in Rzeszow. On the 25th of October she was admitted to Psychiatry Department of the Medical University of Lublin after taking clorazepate as a suicide attempt. While being admitted she was heavily depressed, uttered nihilistic delusions, had suicidal thoughts and was disoriented for time. From etiological point of view the situation can be considered in two categories. On one hand psychotic symptoms may have been the climax of the disturbance of the central nervous system function in long-lasting undiagnosed hypothyroidism. On the other hand, the coincidence of the two disorders cannot be excluded. The presented case implicate the role of the early diagnose of hypothyroidism for the prevention of depression. PMID- 19999805 TI - [Educational standards of the Polish Pharmacoeconomic Society]. AB - The objective of creating Educational Standards of the Polish Pharmacoeconomic Society (Polish Chapter of the International Society for Pharmacoeconomics and Outcomes Research) was to define and establish a scope of knowledge and skills needed for people conducting and interpreting pharmacoeconomic analyses as well as health technology assessments. A Working Group of the Society identified target groups and divided the requirements into 3 groups: basic, intermediate and advanced. The document reflects conditions of the Polish healthcare and educational systems and is harmonized with international regulations in the field of pharmacoeconomics, outcomes research and health technology assessment. The standards may also serve as guidelines for educators in this area. PMID- 19999806 TI - [Platelets haemostatic in patients with multiple myeloma]. AB - Multiple myeloma (MM) is a type of disseminated cancer which derive plasma cells that are immune system cells in bone marrow and produce monoclonal protein. It represents approximately 1% of all cancers, but recent statistics indicate both increasing incidence and earlier age of onset. Haemostatic disorders are an important clinical problem in patients with MM. The most common disturbances symptoms are bleeding and deep-vein thrombosis can due to platelet dysfunction, inhibition of fibrin and increased clearance of coagulation factors connected with monoclonal protein presence. The aim of study is a presentation of disturbances haemostatic in patients with MM, which make up important clinical and diagnostic problems. PMID- 19999807 TI - [Cyclooxygenase inhibitors in chemoprevention and treatment of esophageal squamous cell carcinoma]. AB - Cyclooxygenase inhibitors (COX) are a complex group of pharmacological compounds characterized by significant efficacy in chemoprevention of epithelial origin tumors, especially colorectal ones. It was found that inducible isoform of COX - COX-2 plays an important role in cancer growth and dissemination, e.g., by increase of cellular proliferation, reduction of apoptosis, promotion of local invasiveness and angiogenesis. COX inhibitors decrease prostaglandins' synthesis, but COX-independent mechanisms of their preventive and therapeutical activity were also proven. The influence of COX inhibitors on growth of esophageal squamous cell carcinoma and its precursor lesions was not completely clear. Most studies based on human cancer cell lines revealed antiproliferative and proapoptotic ability of both nonselective (previously called non-steroidal antiinflammatory drugs--NSAIDs) and selective COX-2 inhibitors (coxibs). In in vivo studies performed on animals exposed to chemical carcinogens, the chemopreventive effect was achieved exclusively after administration of experimental selective COX-2 inhibitors, but in the only human trial, supplementation of selective COX-2 inhibitor--celecoxib turned out ineffective. However, many epidemiological data proved effect of prolonged administration of nonselective COX inhibitors, especially acetylsalicylic acid on decreased risk of esophageal squamous cell carcinoma. Few reports concerning application of selective COX-2 inhibitors in patients with invasive squamous cell carcinoma are insufficient for ultimate evaluation of this method of therapy. PMID- 19999808 TI - [Hyperhomocysteinemia and the disturbance of haemostasis--facts and mythes]. AB - The elevated concentration of homocysteine--Hcys (hyperhomocysteinemia) has been correlated with different pathologists, such as cardiovascular diseases. In the article, the some pathways of homocysteine metabolism (i.e. remethylation to methionine, formation of homocysteine thiolactone, and its enzymatic hydrolysis) and the actions of homocysteine and its metabolite--cyclic thioester- homocysteine thiolactone (HTL) on complex process of haemostasis, which regulates the flowing properties of blood, are described. Possible interaction of Hcys and HTL with endothelial cells, blood platelets, plasmatic fibrinogen and other coagulation factors, as the important major components of haemostasis are also discussed. The modification of haemostatic proteins (N-homocysteinylated or S homocysteinylated proteins) induced by Hcys or its thiolactone, seem to be the main reason of biotoxicity of homocysteine in the disturbance of haemostasis and cardiovascular diseases. PMID- 19999809 TI - [Biological function of some elements and their compounds. III. Zinc--component and activator of enzymes]. AB - Zinc plays many basic metabolic functions in human organism as a co-enzymatic or activator about 80 enzymes. It participates in metabolic changes of lipids, proteins and carbohydrates. Zinc easily binds with nucleic acids with beneficial effect on active transport, protein biosynthesis, transcription, and mRNA synthesis. It is inevitable for proper functioning of the respiratory and reproductive systems. Zinc acts protectively in lead and cadmium poisoning and it participates in the elimination of free superoxide radicals (it inhibits of oxygenation of non-saturated fatty acids). It enables normal functioning of the skin and mucous membranes. Zinc deficit can lead to many ailments and disorders of the following systems: muscular, nervous digestive, reproductive, hormonal, immune. PMID- 19999810 TI - [Biological function of some elements and their compounds. IV. Silicon, silicon acids, silicones]. AB - The review is devoted for the occurance, meaning of silicon and their compounds, especially silicon acids and silicones. Silicon participates in biosynthesis of collagen, the basic component of connective tissue. It strengthens and makes the walls of blood vessels more flexible, diminishes capillaries permeability, accelerates healing processes, has a sebostatic activity, strengthens hair and nails. This element has a beneficial effect on phosphorylation of proteins saccharides, and nucleotides. It is also essential for the formation of cytoskeleton and other cellular structures of mechanical or supportive function. Silicon is an initial substrate for obtaining silicones. These are synthetic polymers, in which silicon atoms are bound by oxygen bridges. They are used in almost all kinds of products due to their most convenient physical and chemical properties: moistening and film-forming, giving liquid form increasing solubility. Silicon acids form colloid gel, silica gel, with absorptive abilities, like active carbon. PMID- 19999811 TI - [Burnout as a consequence of chronic stress in the patients suffering from dermatological diseases]. AB - Topic of professional burnout has been discussed in Poland since 80'. Professional burnout is a state of emotional and physical exhaustion caused by excessive and prolonged stress. As the stress continues, you begin to lose the interest or motivation of job, additionally burnout reduces your productivity and energy. That's why it's important to recognize the early signs of burnout and head it off. Three phases of professional burnout have been described: emotional exhaustion, depersonalization and reduced personal accomplishment. The risk of professional burnout involves especially people who are suffering from many disturbances, including dermatological diseases and carry out of professions named Human Services. PMID- 19999812 TI - [Suicide problems--epidemiology, factors, motives and prevention. Part I]. AB - Suicide is a serious public health problem, being, according to WHO, a third leading cause of death among people aged 15-44. In 2001 the total number of suicide deaths was higher than war deaths ( 500.000 and 23.0000 respectively). Due to such a high incidence of suicides it is likely that a doctor may encounter a suicide or attempted suicide. According to NHS data an average GP will most likely have 10-15 suicidal patients during career. A doctor who encounters a suicidal patient or is approached by a worried family member of a suicidal patient should make right decisions and inform a patient as well as his/her family what to do in the future. The aim of this paper is to present basic data regarding epidemiology of suicides and suggestions which may help the doctor to prevent suicide attempts. PMID- 19999813 TI - [Vitamin D--a panacea in nephrology and beyond]. AB - Vitamin D and its active analogues play an essential role in maintaining the calcium and phosphate homeostasis. Vitamin D deficiency may be a consequence of insufficient dietary supply, impaired intestinal absorption, insufficient exposure of the skin to the ultraviolet radiation, or liver and kidney failure. The consequences of mineral metabolism disorders and in particular vitamin D depletion are osteoporosis, increased cardiovascular diseases, immune system impairment and increased morbidity and mortality in patients with end stage of renal disease. These abnormalities are common in patients with chronic kidney disease. Recent clinical studies have shown that vitamin D supplementation in patients with its deficiency, contribute to decreased frequency of bone disorders, cardiovascular incidents, lower risk of several malignancies and to improvement of immune system response regardless of renal function. PMID- 19999814 TI - [Biological active substances of plant origin catechins: dietary sourses, biovailability, the influence on xenobiotic metabolizing enzymes]. AB - Data for monomeric flavan-3-ols (catechins) occurrance and content in foodstuffs of plant origin, as well as the main food sources and dietary intake of these flavonoids are reviewed. The questions on bioavailability, distribution and biotransformation of individual catechins are discussed. The results of in vitro and in vivo investigations of catechins effects on xenobiotic metabolizing enzymes activities were shown. PMID- 19999815 TI - [Influence of specialized carbohydrate protein product enrich vitamins and minerals status of patients with gastrointestianals pathology and osteopenia]. AB - It has been shown that inclusion of the specialized product containing 50% from recommended daily consumption of calcium, 20% of protein, 17-60% of 11 mineral substances and 11 vitamins in the diet of patients suffering from diseases of gastroenteric tract and osteopenia within 6 months lead to increasing of bone mineral density while it has not liquidated the existing vitamin B2 and vitamin D deficiency. The data obtained confirm the expediency of the development of specialized products of the set chemical composition intended for a concrete category of patients, and their inclusion in the diet. PMID- 19999816 TI - [Influence dietary therapy with use sunflower and amaranth oils on parameters of immine reactivity in patients with diabetes mellitus 2 types]. AB - On the basis of the given own researches the positive influence of press non rafinated amaranth and sunflower oils, and as, their mix on immune reactivity on the patients with diabetes mellitus types 2 is shown. PMID- 19999817 TI - [Tongue gustation of patients with arterial hypertension]. AB - To study tongue gustation of patients with arterial hypertension (AH) to sodium chloride (SC) we have examined 630 people. Each one has a gustation threshold to sodium chloride (GTSC). We have established that hypertension patients' GTSC is higher than that of healthy people. Gustation to SC is a changeable index. It depends on nutrition with big (small) SC amount, smoking, current treatment; as to women with overweight, it depends on clinical presentations of AH. PMID- 19999818 TI - [Influence of breads with use of barley, buckwheat and oat flours and barley flakes on postprandial glycaemia in patients with type 2 diabetes mellitus]. AB - It was investigated the influence of breads with use of barley, buckwheat and oat flours and barley flakes on postprandial glycaemia in patients with type 2 diabetes mellitus. It was shown that consumption of breads with use of barley and buckwheat flours is accompanied less marked postprandial glycaemic reaction in compared with standard loading of carbohydrates (wheat bread). Also it was noted greater increase of postprandial glycaemia in consumption of bread with use of barley flakes in compared with consumption of wheat bread inclusive equivalent amount of carbohydrates. PMID- 19999819 TI - [Selenium content in bread of Orenburg region]. AB - Selenium content in wheat and rye bread from 35 districts of Orenburg region has been estimated. The results indicate heterogeneity of selenium distribution and the existence of decreased selenium levels in the West of the region. Different factors affecting selenium accumulation by cereals are discussed: heavy metals pollution, Ni and Cu mining, soil salinization. PMID- 19999820 TI - [Antioxidant in the blood plasmas in population of South Altai in depends an age and sex]. AB - By population of South Altai expose definitely variations in an antioxidant depend in plasma blood on age and sex. PMID- 19999821 TI - [The problems of safety estimation of nanomaterials used in food package]. AB - The main problems related to nanotechnology application in food package and food contact materials are related to absence of identification and migration control methods resource of nanoparticles in food and also to emergency of evaluation of risk from potentially toxic nanoparticles presented in food. The analysis of foreign experience in this field enables the determination of methodological approaches to development of domestic system of food nanotechnology safety. PMID- 19999822 TI - [Content biologically active substance in differences compositions grapes of Dagestan]. AB - The qualitative and quantitative compounds of mineral, phenolic substances and vitamins in grapes of table grades of the average period of the ripening, growing in various ecological conditions is studied. The individual features are revealed. The differentiated approach to selection of grades for use in enotherapy and dietology is offered. PMID- 19999823 TI - [Modern representations of interaction of vitamins]. AB - In article the data of interaction of vitamins among themselves, with various mineral subtances, medicines. The review deals with some new data of the literature on the revealed interactions in manufacture of vitamin complexes, and also in clinicalpractice. The data are systematized according to pharmaceutical, pharmacokinetics and pharmacodynamic interaction of vitamins. PMID- 19999824 TI - [Effect of biologically active supplements on the functional status and physical efficiency of judoists]. AB - A taking of the biologically active supplements by judoists had made a positive influence on the functional status, physical efficiency and oxygen-transporting function of these athletes. An improvement in muscle-strengthening exercises and special endurance has been registered by the athletes, which have taken these biologically active supplements. PMID- 19999825 TI - Measurement and modeling of exposure to selected air toxics for health effects studies and verification by biomarkers. AB - The overall aim of our investigation was to quantify the magnitude and range of individual personal exposures to a variety of air toxics and to develop models for exposure prediction on the basis of time-activity diaries. The specific research goals were (1) to use personal monitoring of non-smokers at a range of residential locations and exposures to non-traffic sources to assess daily exposures to a range of air toxics, especially volatile organic compounds (VOCs) including 1,3-butadiene and particulate polycyclic aromatic hydrocarbons (PAHs); (2) to determine microenvironmental concentrations of the same air toxics, taking account of spatial and temporal variations and hot spots; (3) to optimize a model of personal exposure using microenvironmental concentration data and time activity diaries and to compare modeled exposures with exposures independently estimated from personal monitoring data; (4) to determine the relationships of urinary biomarkers with the environmental exposures to the corresponding air toxic. Personal exposure measurements were made using an actively pumped personal sampler enclosed in a briefcase. Five 24-hour integrated personal samples were collected from 100 volunteers with a range of exposure patterns for analysis of VOCs and 1,3-butadiene concentrations of ambient air. One 24-hour integrated PAH personal exposure sample was collected by each subject concurrently with 24 hours of the personal sampling for VOCs. During the period when personal exposures were being measured, workplace and home concentrations of the same air toxics were being measured simultaneously, as were seasonal levels in other microenvironments that the subjects visit during their daily activities, including street microenvironments, transport microenvironments, indoor environments, and other home environments. Information about subjects' lifestyles and daily activities were recorded by means of questionnaires and activity diaries. VOCs were collected in tubes packed with the adsorbent resins Tenax GR and Carbotrap, and separate tubes for the collection of 1,3-butadiene were packed with Carbopack B and Carbosieve S-III. After sampling, the tubes were analyzed by means of a thermal desorber interfaced with a gas chromatograph-mass spectrometer (GC-MS). Particle-phase PAHs collected onto a quartz-fiber filter were extracted with solvent, purified, and concentrated before being analyzed with a GC-MS. Urinary biomarkers were analyzed by liquid chromatography-tandem mass spectrometry (LC-MS MS). Both the environmental concentrations and personal exposure concentrations measured in this study are lower than those in the majority of earlier published work, which is consistent with the reported application of abatement measures to the control of air toxics emissions. The environmental concentration data clearly demonstrate the influence of traffic sources and meteorologic conditions leading to higher air toxics concentrations in the winter and during peak-traffic hours. The seasonal effect was also observed in indoor environments, where indoor sources add to the effects of the previously identified outdoor sources. The variability of personal exposure concentrations of VOCs and PAHs mainly reflects the range of activities the subjects engaged in during the five-day period of sampling. A number of generic factors have been identified to influence personal exposure concentrations to VOCs, such as the presence of an integral garage (attached to the home), exposure to environmental tobacco smoke (ETS), use of solvents, and commuting. In the case of the medium- and high-molecular-weight PAHs, traffic and ETS are important contributions to personal exposure. Personal exposure concentrations generally exceed home indoor concentrations, which in turn exceed outdoor concentrations. The home microenvironment is the dominant individual contributor to personal exposure. However, for those subjects with particularly high personal exposures, activities within the home and exposure to ETS play a major role in determining exposure. Correlation analysis and principal components analysis (PCA) have been performed to identify groups of compounds that share common sources, common chemistry, or common transport or meteorologic patterns. We used these methods to identify four main factors determining the makeup of personal exposures: fossil fuel combustion, use of solvents, ETS exposure, and use of consumer products. Concurrent with sampling of the selected air toxics, a total of 500 urine samples were collected, one for each of the 100 subjects on the day after each of the five days on which the briefcases were carried for personal exposure data collection. From the 500 samples, 100 were selected to be analyzed for PAHs and ETS-related urinary biomarkers. Results showed that urinary biomarkers of ETS exposure correlated strongly with the gas phase markers of ETS and 1,3-butadiene. The urinary ETS biomarkers also correlated strongly with high-molecular-weight PAHs in the personal exposure samples. Five different approaches have been taken to model personal exposure to VOCs and PAHs, using 75% of the measured personal exposure data set to develop the models and 25% as an independent check on the model performance. The best personal exposure model, based on measured microenvironmental concentrations and lifestyle factors, is able to account for about 50% of the variance in measured personal exposure to benzene and a higher proportion of the variance for some other compounds (e.g., 75% of the variance in 3-ethenylpyridine exposure). In the case of the PAHs, the best model for benzo[a]pyrene is able to account for about 35% of the variance among exposures, with a similar result for the rest of the PAH compounds. The models developed were validated by the independent data set for almost all the VOC compounds. The models developed for PAHs explain some of the variance in the independent data set and are good indicators of the sources affecting PAH concentrations but could not be validated statistically, with the exception of the model for pyrene. A proposal for categorizing personal exposures as low or high is also presented, according to exposure thresholds. For both VOCs and PAHs, low exposures are correctly classified for the concentrations predicted by the proposed models, but higher exposures were less successfully classified. PMID- 19999826 TI - How personal is the political? Democratic revolution and fertility decline. AB - Existing theory has identified the capacity of political revolutions to effect change in a variety of social institutions, although relationships between revolution and many institutions remain unexplored. Using historical data from twenty-two European and four diaspora countries, the author examines the temporal relationship between timing of revolution and onset of fertility decline. The author hypothesizes that specific kinds of revolutionary events affect fertility by engendering ideological changes in popular understandings of the individual's relationship to society and ultimately the legitimacy of couples' authority over their reproductive capacities. Results demonstrate that popular democratic revolutions -- but not institutionalized democratic structures -- predict the timing of the onset of fertility decline. PMID- 19999827 TI - Early scientific progress. PMID- 19999828 TI - Endothelial cells of the meningeal capillaries in the rainbow trout: a freeze fracture study. AB - Meningeal capillaries are unfenestrated. They are made up of endothelial cells that have a pinocytotic caveolae density of 41+/- 11/micron (2) and 89 +/- 21/ micron(2) the abluminal and luminal sides respectively. The total density of intramembrane particles is not significantly different between the luminal and the abluminal membranes;however, the coefficients of partition are significantly different (P < 0.001).One or two strands of tight junctions occur between adjacent cells but no gap junctions nor desmosomes exist. The density of nuclear pores is less than 3.2/micron(2). An abundance of intermediate filaments and free vesicles, some of which are seriated, characterize the cytoplasm. The functional significance of these findings is discussed. PMID- 19999829 TI - R.D. Laing and theology: the influence of Christian existentialism on "The Divided Self". AB - The radical psychiatrist R.D. Laing's first book, "The Divided Self" (1960), is informed by the work of Christian thinkers on scriptural interpretation -- an intellectual genealogy apparent in Laing's comparison of Karl Jaspers's symptomatology with the theological tradition of "form criticism." Rudolf Bultmann's theology, which was being enthusiastically promoted in 1950s Scotland, is particularly influential upon Laing. It furnishes him with the notion that schizophrenic speech expresses existential truths as if they were statements about the physical and organic world. It also provides him with a model of the schizoid position as a form of modern-day Stoicism. Such theological recontextualization of "The Divided Self" illuminates continuities in Laing's own work, and also indicates his relationship to a wider British context, such as the work of the "clinical theologian" Frank Lake. PMID- 19999830 TI - A few laced genes: women's standpoint in the feminist ancestry of Dorothy E. Smith. AB - This article looks at the feminist activism of particular women in the ancestry of the eminent Canadian sociologist, Dorothy E. Smith, and at the archival data that confirm the traces of their influence found in her theory-building. Using the method of interpretative historical sociology and a conceptual framework drawn from Marx called the "productive forces," the article examines the feminist theology of her Quaker ancestor, Margaret Fell, and the militant suffrage activism of her mother and her grandmother, Dorothy Foster Place and Lucy Ellison Abraham, respectively. The article argues that the household labour of the remarkable women in her family line became a "productive force" that facilitated her imagining of the feminist theory, "the standpoint of women". PMID- 19999831 TI - Meeting of the Strategic Advisory Group of Experts on immunization, October 2009 conclusions and recommendations. PMID- 19999832 TI - Polynesia and polygenism: the scientific use of travel literature in the early 19th century. AB - Christoph Meiners (1747-1810) was one of 18th-century Europe's most important readers of global travel literature, and he has been credited as a founder of the disciplines of ethnology and anthropology. This article examines a part of his final work, "Untersuchungen uber die Verschiedenheiten der Menschennaturen" [Inquiries on the differences of human natures], published posthumously in the 1810s. Here Meiners developed an elaborate argument, based on empirical evidence, that the different races of men emerged indigenously at different times and in different places in natural history. Specifically this article shows how a sedentary scholar who never left Europe constructed a narrative of human origins and migrations on the basis of (1) French theory from the 1750s (Charles de Brosses and Simon Pelloutier) and (2) data gathered by explorers as reported in travel literature (J.R. Forster, Perouse, Cook, Marsden). PMID- 19999833 TI - The social, cosmopolitanism and beyond. AB - First, this article will outline the metaphysics of "the social" that implicitly and explicitly connects the work of classical and contemporary cosmopolitan sociologists as different as Durkheim, Weber, Beck and Luhmann. In a second step, I will show that the cosmopolitan outlook of classical sociology is driven by exclusive differences. In understanding human affairs, both classical sociology and contemporary cosmopolitan sociology reflect a very modernist outlook of epistemological, conceptual, methodological and disciplinary rigour that separates the cultural sphere from the natural objects of concern. I will suggest that classical sociology -- in order to be cosmopolitan -- is forced (1) to exclude "non-social and non-human objects" as part of its conceptual and methodological rigour, and (2) consequently and methodologically to rule out the non-social and the non-human. Cosmopolitan sociology imagines "the social" as a global, universal "explanatory device" to conceive and describe the non-social and non-human. In a third and final step the article draws upon the work of the French sociologist Gabriel Tarde and offers a possible alternative to the modernist social and cultural other-logics of social sciences. It argues for a inclusive conception of "the social" that gives the non-social and non-human a cosmopolitan voice as well. PMID- 19999834 TI - Organ transplantation in Saudi Arabia. PMID- 19999835 TI - University challenge. PMID- 19999836 TI - Investment, recruitment and support: initiatives to tackle the midwifery crisis. PMID- 19999837 TI - A lifeline for children. AB - CLIC Sargent, the charity for children with cancer, is next year's partner for the London Marathon. Raising money from the event will fund more nurses to provide care. PMID- 19999838 TI - Discover your bias. AB - Online questionnaires uncover people's prejudices. PMID- 19999839 TI - Action on emissions. AB - More than 40 NHS organisations have signed up to reduce their carbon footprints by 10 per cent in 2010 as part of the 10:10 campaign. PMID- 19999840 TI - Learn to say 'no'. PMID- 19999841 TI - Medicines management. PMID- 19999842 TI - Management of procedures and reactions following first aid. AB - Registered nurses may fear that offering first aid will expose them to legal action and emotional stress. The final article of this series emphasises the importance of accurate record-keeping and provides advice on co-operating with legal procedures, including providing information to other professionals, obtaining consent and reporting a death. Providing emergency first aid can be an emotional experience. This article offers advice on coping after the incident, and reflection as a means of allowing nurses to assess their actions and resolve any issues. PMID- 19999843 TI - Risk factors for moving and handling bariatric patients. AB - AIM: To identify and explore the manual handling risks for patients and caregivers using bariatric patient pathways in health and social care. METHOD: A mixed methodological approach, including focus groups and questionnaires, was used in this study. Participants were recruited for the focus groups from the National Back Exchange (NBE) Special Interest Group on Bariatrics and the National Ambulance Risk and Safety Forum (NARSF). The questionnaire was distributed to all members of the NBE and NARSF in 2006. RESULTS: The data sets were analysed thematically (focus groups: n = 25) and descriptively (questionnaires: n = 230). Patient pathways were mapped for medical (acute), community and maternity admissions resulting in five generic themes: patient factors; building and vehicle space and design; equipment and furniture; communication; and organisational and staff issues. A total of 59% (n = 136) of respondents reported that their organisation did not have a bariatric manual handling policy. Of all responding organisations (n = 230), 77% (n = 177) had access to specialist equipment but only 32% (n = 68) of NBE respondents (n = 212) provided specialist bariatric manual handling training for staff. Lack of formal communication systems between and within organisations contributed to manual handling risks at admission and discharge. CONCLUSION: There is a need to review and design more appropriate buildings, vehicles, furniture and equipment to reduce the manual handling risks to health professionals and bariatric patients. More research is required to examine the implementation of policies, in particular focusing on the use of equipment and the communication between care providing agencies. PMID- 19999844 TI - Role modelling practice with students on clinical placements. AB - This article explores practical ways of role modelling practice with students on clinical placements. Students are frequently assigned to a senior practitioner for periods of observation, which involves shadowing the practitioner in practice. Learning opportunities are not necessarily seized because a strategy is not in place to enhance learning. The article provides guidance on how to proceed effectively with role modelling. PMID- 19999845 TI - Understanding haematuria. PMID- 19999846 TI - Beyond the call of duty. PMID- 19999847 TI - New act, new ethos, new role. PMID- 19999848 TI - Pioneer's legacy of change. PMID- 19999849 TI - To test or not to test: parent information for discussions of food allergy and autism. PMID- 19999850 TI - Advanced primary medical care 2025. PMID- 19999851 TI - Living wills and a medical treatment powers of attorney. PMID- 19999852 TI - On-site assessments recognize medical practices that excel in risk management. PMID- 19999853 TI - "Physician heal thyself". A look at stress management, physical wellness and addiction recovery. PMID- 19999855 TI - How to recruit a J-1 Visa candidate to your practice. PMID- 19999854 TI - Physician-focused recovery program changes lives. PMID- 19999856 TI - Flu Season 101: make sure all staff are vaccinated with seasonal flu vaccine. PMID- 19999857 TI - Hib booster dose for children ages 12-15 months reinstated. PMID- 19999858 TI - To young physicians: why Michigan is your future. PMID- 19999859 TI - The dollars & sense of health information technology (HIT) funding. PMID- 19999860 TI - Hybrid endovascular repair of thoracic aortic aneurysms by debranching and the creation of landing zone-zero. AB - Thoracic aortic aneurysms (TAA) have remained a formidable operative challenge. Open surgical techniques have been associated with high rates of morbidity and mortality. Thoracic endovascular aneurysm repair (TEVAR) has produced results equal to or better than the traditional open surgical approach. This report presents a patient with a complex thoracic aortic aneurysm involving the ascending, transverse, and proximal descending thoracic aorta. This patient was successfully managed by the creation of Landing Zone-Zero, arch vessel debranching, and endografting the entire aortic arch without the need for hypothermic circulatory arrest or cerebral perfusion strategies. Computer tomographic images demonstrate the repair to be durable at 18 months. PMID- 19999862 TI - Kentucky pilot project for primary PCI without onsite CABG. AB - PURPOSE: The American College of Cardiology, the American Heart Association, and the Society for Cardiovascular Angiography and Interventions strongly recommend that primary Percutaneous Coronary Intervention (PCI) should be performed in facilities that have an experienced cardiovascular surgical team available as emergency backup for all procedures. The current study investigates the medical soundness of allowing select facilities in Kentucky to perform primary PCI despite being devoid of onsite emergency backup capabilities. METHODS: Two hospitals in the state of Kentucky, currently without emergency backup capabilities, have been allowed to perform primary PCIs for more than three years (beginning in April 2005) by the Kentucky Cabinet for Health and Family Services. The two hospitals selected were of similar size (approximately 200 beds) and similar distances from hospitals with onsite emergency backup capabilities (approximately one hour). We performed an analysis evaluating if hospitals without backup surgical capability have similar outcomes when compared to hospitals with backup surgical capabilities. Outcome variables included: (1) mortality, (2) cardiac arrest as result of PCI, (3) emergency surgery performed as a result of PCI, and (4) door-to-balloon time. RESULTS: Our results suggest that there is no significant difference in any of the outcome variables studied between facilities with and without onsite emergency backup capabilities. CONCLUSIONS: Recommendations concerning primary PCI may need to be revisited. The principal outcomes associated with primary PCI were not significantly affected by whether a facility has onsite emergency backup capabilities. Therefore, we recommend that hospitals without backup surgical capabilities be allowed to perform primary PCI (with restrictions based on surgeon experience and the facilities' volume). PMID- 19999861 TI - An investigation of an outbreak: Methicillin resistant Staphylococcus aureus colonization in an intensive care cardiac unit. AB - BACKGROUND: Colonization in the nares with Methicillin Resistant Staphylococcus aureus (MRSA) has been described as a risk factor for eventual MRSA infection. The objective of this report is to describe the outbreak investigation, to identify the risk factors involved, and to evaluate and recommend control measures. METHODS: This study was a retrospective observational outbreak study carried out in the Intensive Care and Cardiac Unit. Percentages of patients with positive conversions were graphically plotted out and were used to determine expected percentage of conversions versus observed. The case definition was determined to be a patient with a positive culture for MRSA from a nasal swab upon discharge or transfer from the ICCU whose initial swab on admission was negative. RESULTS: The expected number of conversions was 2% per month and the newly observed number of conversions was 6%, which was >3 standard deviations from the mean. Repetitive sequence-based PCR (rep-PCR) was performed for the microbial DNA typing, which discovered genetically identical strains. DISCUSSION: The main finding in this outbreak investigation was that the common room which housed four of these patients was contaminated with MRSA. A limitation was that some patients who would have been included in the study did not get screened at discharge and their conversion status could not be ascertained and environment cultures were not performed. Active surveillance allowed for the detection of an outbreak of MRSA colonization which led to early intervention that prevented more patients from becoming colonized. Prevention of colonization should be one of the primary goal in the prevention of MRSA. PMID- 19999863 TI - The media, substance dependence, and misinformation. PMID- 19999864 TI - Abundance--the dilemma. PMID- 19999865 TI - Increasing the medical school application pool: a key to training more rural physicians. PMID- 19999866 TI - Foot compartment syndrome: a systematic review of the literature. AB - We systematically reviewed published evidence regarding foot compartment syndrome with regards to causes, methods of diagnosis, number of incisions used for fasciotomy, wound closure techniques, complications, and functional outcomes. Publications were collected using PubMed and OVID databases, and were reviewed as above. All were retrospective case series (evidence-based medicine level IV). Four articles with 39 cases of foot compartment syndrome were reviewed in all. The most common cause of foot compartment syndrome was crush injury to the foot. Diagnosis was mostly made through a combination of clinical findings and compartment pressure measurements. Sixty-five percent of cases required split thickness skin grafts for wound closure after fasciotomy. Neurological deficits were the most common complication (52%). Thirty-nine percent of the patients reported residual pain and stiffness while ten percent could return to work or their pre-injury activity state after fasciotomy. PMID- 19999867 TI - Open distal clavicle resection: isolated or with adjunctive acromioplasty. AB - The purpose of this study was to assess outcomes following open distal clavicle resection for acromioclavicular joint arthritis or distal clavicle osteolysis, with and without associated acromioplasty. Patients with painful clinical findings limited to the acromioclavicular joint had isolated distal clavicle excision (23 shoulders). Patients with acromioclavicular joint abnormalities and rotator cuff tendinopathy also underwent acromioplasty (41 shoulders). At average follow-up of 8.3 years, pain scores improved from 4.7 (1 to 5 scale) to 2.3 (p < 0.001). Patient satisfaction improved from 1.8 (1 to 10 scale) to 8.3 (p < 0.001). Postoperatively the mean Simple Shoulder Test (SST) score was 10.9. The mean American Shoulder and Elbow Surgeons (ASES) Score was 88.3. There were no statistical differences in pain, satisfaction, motion, and shoulder scores between the two groups. Results of distal clavicle resection with or without acromioplasty are favourable with a low rate of complications and seldom is further surgery required. PMID- 19999868 TI - A comparative clinical evaluation of arthroscopic single-row versus double-row supraspinatus tendon repair. AB - Cadaveric studies and commercial pressure have initiated a strong trend towards double-row repair in arthroscopic cuff surgery. The objective of this study was to evaluate if the biomechanical advantages of a double-row supraspinatus tendon repair would result in superior clinical outcome and higher abduction strength. A retrospective study of two groups of 32 single-row and 33 double-row repairs of small to medium cuff tears was performed. The Simple Shoulder Test (SST) and a visual analog scale for pain were used to evaluate the outcome. The participation rate was 100%. A subset of patients was further investigated with the Constant Score (CS) including electronic strength measurement. The double-row repair patients had significantly more (p = 0.01) yes answers in the SST than the single row group, and pain reduction was slightly better (p = 0.03). No difference was found for the relative CS (p = 0.86) and abduction strength (p = 0.74). Patient satisfaction was 100% for double-row and 97% for single-row repair. Single- and double-row repairs both achieved excellent clinical results. Evidence of superiority of double-row repair is still scarce and has to be balanced against the added complexity of the procedure and higher costs. PMID- 19999869 TI - Relocation of a dislocated long head of biceps tendon is no better than biceps tenodesis. AB - A displaced Long Head of Biceps (LHB) tendon is commonly encountered in association with subscapularis rotator cuff tears. Management options for the displaced tendon consist of tenotomy, tenodesis or relocation with reconstruction of the biceps pulley. We present 16 patients in whom LHB relocation and reconstruction of the biceps pulley, was performed in association with subscapularis rotator cuff repair. During follow-up ultrasound scanning was used to assess LHB mobility and location. Eight of the 16 patients had a static LHB tendon at an average follow-up of 26 months. Four of the 6 patients who had a groove deepening procedure combined with the tendon relocation had a static tendon on ultrasound scanning. Relocation of the LHB and reconstruction of the biceps pulley appears to offer no advantage over tenotomy or tenodesis alone when managing the displaced LHB tendon in conjunction with subscapularis tears. PMID- 19999870 TI - Retrograde Ender nailing for humerus shaft fractures. AB - Humerus fractures are common; their management remains controversial. Infection, neurovascular injury, joint problems and non-union are recognised complications of surgical treatment. These complications can be decreased by opting for a surgical treatment that is less invasive and safe. We present a series of 59 patients treated with retrograde Ender nailing; 56 healed in an average of 9.1 weeks, 2 had delayed union (> 15 weeks) and one went on to non-union, which healed after secondary plate fixation. Nail back out occurred in 8 cases, of which only 3 required nails repositioning. The mean Constant score at final follow-up (mean 19 months) was 91; it was significantly lower in patients over 50 years of age and in those with segmental fractures. In this series, Ender retrograde nailing gave overall satisfactory results and appeared as a safe and efficient technique. PMID- 19999871 TI - Outcome of ulnohumeral arthroplasty in osteoarthritis of the elbow. AB - Ulnohumeral arthroplasty is commonly used to treat mild or moderate osteoarthritis (OA) of the elbow. Previous studies have reported good to excellent results, but have not focused on the continuing degenerative process which can alter the outcome. The authors retrospectively evaluated 10 patients with elbow OA who were treated with the Outerbridge-Kashiwagi procedure from 2004 to 2006. The mean increase in the overall Andrews and Carson scores was 91 points (range: 70 to 100) at 3 months followup. The overall gain in range of motion, patient satisfaction rate and VAS and Andrews and Carson scores was found to be significantly diminished at the last follow-up, as compared with early postoperative values. Although ulnohumeral arthroplasty is an accepted procedure for elbow osteoarthritis, the deterioration of the overall gain in pronation/supination and progression of the disease can affect the final scores and patient satisfaction. PMID- 19999872 TI - Treatment of non-union of forearm bones with a free vascularised corticoperiosteal flap from the medial femoral condyle. AB - We report 6 cases of recalcitrant non-union of forearm bones treated with a free vascularized thin corticoperiosteal graft harvested from the medial condyle of the femur. All non-unions healed. In one 58-year-old patient a fracture of the femur occurred. PMID- 19999873 TI - Carpal scaphoid fracture in the skeletally immature: a single centre one-year prospective study. AB - We report up-to-date one-year prospective data on the incidence of scaphoid fracture in skeletally immature patients managed in a Fracture Clinic of a Level I Trauma Centre in a University Hospital in the UK. All scaphoid fractures were immobilised until union was achieved. One hundred twenty one skeletally immature patients were referred for a suspected fracture of the scaphoid. Fourteen patients (11.5%) did have a scaphoid fracture, an incidence of 15 per 100,000 (0.55% of all paediatric injuries referred). Mean delay in being seen was 2.6 days, and mean follow-up time was 52.3 days. Only plain radiography was used to diagnose and follow up scaphoid fractures. Fractures of the scaphoid in skeletally immature individuals are uncommon, are usually undisplaced, occur more commonly in the distal portion of the bone, and carry a good prognosis. PMID- 19999874 TI - Clinical and radiological evaluation of modular trabecular metal acetabular cups. Short-term results in 64 hips. AB - Bone ingrowth and long-term survival of cementless acetabular implants, both in primary and revision hip arthroplasty, are determined by initial implant stability and by the osteoconductive properties of the metal shell. The outcome of a trabecular metal tantalum modular uncemented cup was evaluated in 64 hips in 62 patients. Eleven cups were used in complex primary total hip replacements, 53 cups were implanted in revision hip arthroplasties. The follow-up ranged from 12 months to 48 months. Clinically the Charnley modified Merle-d'Aubigne Postel score improved from 9 to 16 in the primary arthroplasty group and from 10 to16 in the revision group. Serial radiographs demonstrated excellent stability and bone apposition; in the revision cases, graft incorporation was noted in all but one cup. The number of periacetabular gaps detected on initial postoperative radiographs in the revision cases decreased from 15 to 4 and none of the remainder deteriorated with time. Complications included 2 haematomas, 1 dislocation and 1 cup failure in a Paprosky IIIb defect because of cup undersizing. The early results with the trabecular metal modular cementless cup appear promising in both complex primary and revision hip arthroplasty, even in the presence of considerable bone loss that requires additional bone grafting. PMID- 19999875 TI - Endoscopically assisted core decompression in avascular necrosis of the femoral head. AB - The authors have used bone endoscopy (medulloscopy) to improve the accuracy of the conventional technique for core decompression and grafting in 36 hips with avascular necrosis of the femoral head. Endocopy was found to facilitate debridement of the necrotic bone and to decrease the risk of perforation of the cartilage during debridement. The endoscopy-assisted technique failed, however, to improve the final outcome of core decompression and grafting in more advanced stages of osteonecrosis. Based on this experience, the authors do not recommend core decompression, with or without endoscopic assistance, in advanced stages of avascular necrosis, but they suggest using endoscopic assistance during decompression in small-size lesions. PMID- 19999876 TI - Combined syringe cement pressurisation and intra-osseous suction: an effective technique in total knee arthroplasty. AB - Our study looked at the short and medium-term results of a cost-effective technique of bone surface preparation and cement introduction in total knee arthroplasty (TKA). Numerous factors determine the outcome following TKA. Early failure and component loosening have been attributed to inadequate bone-cement and prosthesis-cement interfaces, established at the time of surgery. Various operative techniques have been reported to achieve effective cement penetration and inter-digitation into cancellous bone, with techniques of cement pressurization and intra-osseous suction being employed. We have devised a technique of cement pressurization using a modified standard 20 ml syringe, combined with intra-osseous suction. Retrospective evaluation of a series of 50 post-operative radiographs has shown that effective and even penetration of cement to a depth of 8.0 to 10.6 mm can be achieved consistently with this technique, during TKA, without the use of tourniquet. In addition evaluation of post-operative plain radiographs with minimum follow-up of 5 years showed 16 knees with minor, non-progressive lucent lines around the tibial component with a maximum Knee Society TKA Roentgenographic score of 2. To conclude we propose that this simple technique of surface preparation and cement introduction produces good results in the short and medium-term. PMID- 19999877 TI - The LISS plate treatment of supracondylar fractures above a total knee replacement: a case-control study. AB - The aim of this study was to assess the outcome of periprosthetic fractures above a total knee arthroplasty treated by LISS plate fixation. The primary outcome measure was the radiological union. Secondary outcome measures were knee function using the Oxford Knee Score, and quality of life using the SF-12 questionnaire. Over a 4-year period we treated 16 supracondylar periprosthetic fractures with a LISS plate in 15 patients. Eleven patients (12 fractures) were followed up for an average of 8 months. Eleven out of these 12 fractures went on to unite in an average of 3.7 months. Functional outcome questionnaires were filled out by 9 patients. Suitable matches were found for a subgroup of 5 patients (6 LISS plates). No statistically significant differences were found in the Oxford knee scores (p = 0.749) or quality of life scores (p = 0.440) between the two groups. Our data suggest favourable outcomes after LISS plate fixation of supracondylar periprosthetic femoral fractures above total knee replacement. PMID- 19999878 TI - Cutaneous sensory loss following primary total knee arthroplasty. A two years follow-up study. AB - We examined 32 patients for skin flap numbness around their scars after primary total knee arthroplasty (TKA). All 32 patients were reviewed six weeks following surgery and two years thereafter. The aims of our study were to determine the natural history of cutaneous sensory loss, to identify the relationship with scar length and scar location, as well as the relationship between the size of the numb area and the severity of numbness and its recovery pattern. We also studied the relationship between numbness and other factors such as tourniquet time, lateral release and patella resurfacing. Twenty-six patients (81%) had lateral skin flap numbness; the other six patients (19%) had normal skin sensation around the scar. The size of the numb area was large in 19 patients (73%) out of 26. Our findings suggest more severe sensory loss is associated with larger numb area. Patients with a smaller numb area had less severe sensory loss. More laterally placed incisions were having better sensation (p = 0.00435). Patella resurfacing (p = 0.5) and lateral retinacular release (p = 0.10) were not significantly associated with the skin numbness. Fifty percent of our patients fully recovered from skin numbness at the end of two years. Patients with a small numb area had a more favourable outcome compared to those with a larger numb area. We conclude that permanent cutaneous sensory loss is not a universal occurrence following primary TKA; however a significant number of patients can be affected by this complication. All the patients should be warned about cutaneous numbness before the surgery. PMID- 19999879 TI - Plantar contact stress and gait analysis after resection of tarsal coalition. AB - The purpose of this study was to assess the foot loading characteristics and foot function of patients after operative correction of a tarsal coalition. Ten patients who had undergone operative treatment of a tarsal coalition were included in this study. One foot was affected in five patients and both feet in the other five. A calcaneonavicular coalition was present in 12 feet and a talocalcaneal coalition in three feet. Mean follow-up was 11.3 years. Clinical evaluation was based on a standardized questionnaire, a visual analogue scale for pain (VAS), the American Orthopaedic Foot and Ankle Society (AOFAS) Score and radiographic evaluation of the last radiographs. An objective analysis of foot loading characteristics was carried out with instrumented gait analysis and pedobarography. The clinical results were overall fair for pain, range of motion and walking distance. The AOFAS also showed fair results (mean: 78.1) at follow up. Gait analysis revealed alterations in kinematic and kinetic parameters for the operated foot. Pedobarographic analysis showed altered loadings for heel and forefoot. In this study, operative treatment of tarsal coalition achieved fair clinical and radiographic results and did not restore physiologic gait and foot loading. PMID- 19999880 TI - Treatment of hallux valgus deformity: preliminary results with a modified distal metatarsal osteotomy. AB - This study aimed to investigate the preliminary results achieved with a modified distal first metatarsal osteotomy (reversed L-shaped) for correction of moderate and severe hallux valgus deformities. This prospective study included 31 patients (39 feet) with a mean age of 56 years. All patients underwent a reversed L-shaped osteotomy of the first metatarsal. At follow-up all patients were reviewed clinically and radiologically. Patients were categorized into two groups (MTP angle A: < 20 degrees, and B: 20-40 degrees). The AOFAS score improved from 53 points to 91 points at follow-up (p < 0.0001). Group A showed an increase from 56 to 90 points (p = 0.003), group B from 52 to 92 points (p < 0.0001). The mean 1-2 inter-metarsal angle (IMA) decreased from 12.5 degrees preoperatively to 8 degrees at follow-up (p < 0.005): from from 11 degrees to 9 degrees in group A (p = 0.09) and from from 13 degrees to 7 degrees in Group B (p < 0.0001). No nonunion or avascular necrosis was observed. One diabetic patient developed a resistant postoperative infection. The L-shaped osteotomy provided good and excellent clinical as well as radiological results in the mild and moderate-to severe hallux valgus deformities treated. The mid- and long-term effect of this type of osteotomy needs to be further investigated. PMID- 19999881 TI - Intra-operative cerebral microembolisation during primary hybrid total hip arthroplasty compared with primary hip resurfacing. AB - Fat embolism during total joint arthroplasty or intramedullary procedures is well documented and is infrequently fatal. Considerable morbidity is associated with fat embolism syndrome, and post operative cognitive dysfunction is frequently seen, yet the exact pathophysiology remains unclear. Intraoperative cerebral microemboli can be detected using transcranial Doppler ultrasound and moreover the presence of a patent foramen ovale (PFO) may be examined for using a validated technique employing this modality. Persistent patent foramen ovale may act as a conduit for embolic material to traverse from the venous to the systemic circulation and consequently affect cerebral function. We wished to 1) investigate the incidence of cerebral microembolisation during primary hybrid total hip arthroplasty and compare this with hip resurfacing, 2) examine the influence of patent foramen ovale on cerebral microembolisation and 3)assess the influence of cerebral microemboli on the outcome of patients undergoing these procedures. We prospectively compared 12 patients undergoing hip resurfacing with 12 patients undergoing hybrid total hip replacement (THR) for the incidence and load of intraoperative cerebral microemboli, using transcranial Doppler. All patients were tested for the presence of a patent foramen ovale using a validated technique. Outcome was assessed using the WOMAC, Harris Hip Score, Oxford Hip Score and EuroQoL quality of life measure. No patient in the hip resurfacing group demonstrated intra-operative cerebral microembolisation. Five patients in the THR group showed transcranial Doppler evidence of microemobli during the procedure. With the small numbers of patients available, there was not a significant difference in microemboli load between the groups (p = 0.09). There was no significant difference between the groups regarding the incidence of PFO (p = 0.78). There was no significant relationship between the incidence and total microemboli load and the incidence of PFO (p = 0.56). There was no significant difference in outcome at six months between patients who demonstrated microemboli and those who did not. The incidence of cerebral microembolisation during hip resurfacing appears to be very low. Although our study demonstrated cerebral microemboli in a significant proportion of patients undergoing primary hybrid THR, the numbers of microemboli were low and the presence of a patent foramen ovale did not influence microemboli incidence or load. Finally, patients who demonstrated cerebral microemboli did not have a worse outcome than patients who did not. PMID- 19999883 TI - Glomus tumour of the deltoid muscle. A case report. AB - Glomus tumours around the shoulder are very rare. To the best of our knowledge, seven cases have been reported to date. We present a case with special emphasis on its localization and time of appearance. Contrary to the related reports in the literature, the lesion was of short duration and it became symptomatic just 6 months before referral. Complete pain relief was achieved with surgical treatment. PMID- 19999882 TI - Arthroscopic fenestration of the distal humerus: a viable option for painful elbow impingement in sportsmen. AB - Open ulnohumeral arthroplasty, also known as the Outerbridge-Kashiwaghi procedure, is performed in elbow arthritis to relieve pain and improve the range of motion. A similar technique of distal humeral fenestration is applied in elbow arthroscopy to achieve pain relief in degenerative elbow arthritis. We report a possible new indication in young sportsmen with recurrent posterior elbow impingement. A professional javelin thrower and a basketball player with recurrent loose bodies, posterior impingement and a minimal extension lack were free of complaints and resumed all sports activities within 6 weeks after an arthroscopic fenestration of the distal humerus. Their elbow function remained perfect within a 2 year follow-up period. We suggest that arthroscopic ulnohumeral arthroplasty is a viable option in sportsmen with recurrent loose bodies and elbow locking due to ongoing elbow arthritis. PMID- 19999884 TI - Aneurysmal bone cyst of the scapula. A case report. AB - Aneurysmal bone cyst (ABC) is an uncommon, benign but locally destructive bone lesion of unknown origin. Differential diagnosis can be challenging as it shares common radiological and pathological features with other benign and malignant bone lesions. The degree of diagnostic difficulty grows even more when an unusual location has to be taken into account. We report a rare and challenging case of a large primary ABC located at the scapula of a young male, who was surgically treated with subtotal removal of the scapula. PMID- 19999885 TI - Tumoral calcinosis in children, a challenging and possibly underdiagnosed condition. AB - Tumoral calcinosis is a rare condition characterized by deposits of calcium hydroxyapatite crystals in periarticular soft tissues. Three clinical settings are possible: complication of renal dialysis, hereditary and sporadic. The condition more commonly affects adults, is rare in children and extremely uncommon in infants. A case of sporadic tumoral calcinosis of the hip is reported in a six-year-old boy for whom the diagnosis was challenging. Surgical treatment was applied because of pain and major functional impairment. A pharmacologic treatment was added for two years. After three years of follow-up, the child was completely asymptomatic and had regained full range of motion. The diagnosis of tumoral calcinosis in children remains challenging. PMID- 19999886 TI - Secondary collapse of lateral femoral condyle following bone bruise: a case report. AB - The term bone bruise, in modern medicine, is still a relatively young one, which followed the advent of MRI. Resolution takes between 12 and 24 weeks, however clinical significance is unknown. We present the case of an 18-year-old male who developed bone bruising of his lateral femoral condyle, associated with meniscal injury and anterior cruciate ligament rupture, following a fall from a push bike. A subsequent injury then led to collapse of his lateral femoral condyle following initial resolution of his symptoms. This was managed operatively performing bony and soft tissue stabilisation. This case highlights the issues associated with proven bone bruising and associated soft-tissue injuries in the knee, which cannot be considered innocuous injuries. We also raise the question as to whether patients should undergo a period of protected weightbearing when a bone bruise is recognised on MRI. PMID- 19999887 TI - Traumatic talus extrusion: case reports and literature review. AB - Total talar dislocation is a rare injury, which is usually open and with associated fractures. We report two cases of open anterolateral talar extrusions following high-energy traffic accidents. Both were treated with wound debridement, joint irrigation, closed reduction and external fixation. At 12 months follow-up, both patients had pain-free motion of the ankle without any signs of avascular necrosis or arthritis. PMID- 19999888 TI - Giant sacral schwannoma: a case report and review of the literature. AB - Intraspinal schwannomas localized in the sacrum are relatively infrequent, accounting for 1-5% of all spinal axis schwannomas. They frequently grow to considerable size before detection; hence, the term giant sacral schwannoma. Sacral schwannomas arise from the sacral nerve roots. The diagnosis of schwannomas in the spinal canal is difficult because of their slow growth, often resulting in extensive bony destruction. This case report documents the management of a 48-year-old male with a giant sacral schwannoma. We performed a two-stage surgery with intralesional tumour resection. The patient is now free of any complaint, complications and there is no recurrence two years after resection of the schwannoma. Intralesional excision of a sacral schwannoma is a less invasive procedure than total or partial sacrectomy. Using a combined anterior and posterior approach, satisfactory tumour excision and stabilization can be achieved, while avoiding the high morbidity related with total sacrectomy. PMID- 19999889 TI - Medication trials in an imperfect world: gout and Parkinson's disease. PMID- 19999890 TI - An ailment for the royally nourished. PMID- 19999891 TI - Introduction: hyperuricemia and gout. PMID- 19999892 TI - Medical implications of hyperuricemia. PMID- 19999893 TI - Diagnosis and management of acute gout. AB - A definitive diagnosis of acute gout is made by detection of monosodium urate crystals in the synovial fluid of an inflamed joint. However, when this is not feasible a clinical diagnosis can sometimes be made with reasonable accuracy. The mainstays of acute gout management are colchicine, NSAIDs, and systemic or intra articular corticosteroids. NSAIDs are preferable to colchicine because of their more favorable side effect profile. Successful treatment occurs with the prompt initiation of high dose short half-life NSAIDS. Since many patients with gout have comorbidites that preclude the use of NSAIDS or colchicine, systemic corticosteroids are commonly used to treat acute gouty arthritis. Intra-articular injections are appropriate in the setting of mono- or oligoarticular involvement. Adequate duration of anti-inflammatory therapy and careful patient education are essential elements of successful therapy for acute gout. Evaluation and management of hyperuricemia should be undertaken when all symptoms of acute gout are resolved and the patient is stable on daily prophylaxis with NSAIDs or colchicine. PMID- 19999894 TI - Approach to the treatment of hyperuricemia. PMID- 19999895 TI - Gout in women. PMID- 19999897 TI - Intranasal mucosal malignant melanoma. PMID- 19999896 TI - Treatment failure gout. PMID- 19999898 TI - Use of angiotensin-converting enzyme inhibitors/angiotensin receptor blockers and lipid-lowering therapies among Rhode Islanders with diabetes enrolled in Medicare Part D plans in 2006 and 2007. PMID- 19999900 TI - The wanderings of the vagus nerve. PMID- 19999899 TI - Asymptomatic versus symptomatic urinary tract infections in long-term-care facility residents. PMID- 19999901 TI - Diabetes prevention and control: progress towards healthy people 2010 goals. PMID- 19999902 TI - Prevention of relapsing mediocrity: how to maintain performance improvement in hospitals. PMID- 19999903 TI - Management of bile duct injuries: a practical approach. AB - The incidence of bile duct injury (BDI) has increased after the introduction of laparoscopic cholecystectomy. A BDI can occur in the hands of experienced surgeons also. It can result in serious complications and may even cause death of the patient; it also has financial and legal implications. Proper training, sound surgical technique, and conversion to an open operation can prevent a large number of injuries. An injury that is missed during the operation manifests in the postoperative period as a bile leak and external biliary fistula or during the follow up as a biliary stricture. Management of a BDI depends on the nature of the bile duct injured, type of injury, and expertise available; it may range from simple subhepatic drainage to Roux-en-Y hepaticojejunostomy. Excellent results can be obtained when BDI is managed at a hepatobiliary center. PMID- 19999904 TI - Carolinas Medical Center surgical teaching program. PMID- 19999905 TI - A central venous line protocol decreases bloodstream infections and length of stay in a trauma intensive care unit population. AB - We evaluated the benefit of a central venous line (CVL) protocol on bloodstream infections (BSIs) and outcome in a trauma intensive care unit (ICU) population. We prospectively compared three groups: Group 1 (January 2003 to June 2004) preprotocol; Group 2 (July 2004 to June 2005) after the start of the protocol that included minimizing CVL use and strict universal precautions; and Group 3 (July 2005 to December 2006) after the addition of a line supply cart and nursing checklist. There were 1622 trauma patients admitted to the trauma ICU during the study period of whom 542 had a CVL. Group 3 had a higher Injury Severity Score (ISS) compared with both Groups 2 and 1 (28.3 +/- 13.0 vs 23.5 +/- 11.7 vs 22.8 +/- 12.0, P = 0.0002) but had a lower BSI rate/1000 line days (Group 1: 16.5; Group 2: 15.0; Group 3: 7.7). Adjusting for ISS group, three had shorter ICU length of stay (LOS) compared with Group 1 (12.11 +/- 1.46 vs 18.16 +/- 1.51, P = 0.01). Logistic regression showed ISS (P = 0.04; OR, 1.025; CI, 1.001-1.050) and a lack of CVL protocol (P = 0.01; OR, 0.31; CI, 0.13-0.76) to be independent predictors of BSI. CVL protocols decrease both BSI and LOS in trauma patients. Strict enforcement by a nurse preserves the integrity of the protocol. PMID- 19999906 TI - Is there a benefit to multidisciplinary rounds in an open trauma intensive care unit regarding ventilator-associated pneumonia? AB - Multidisciplinary rounds (MDRs) have been instituted for patient care since June 2005. Before June 2005, all care was provided by individual practitioners. MDRs include the surgical intensivist, surgical resident, patient's nurse, case manager, pharmacist, chaplain, nutritionist, and respiratory therapist. Our study examined the effect of MDRs on ventilator-associated pneumonia in trauma patients in open intensive care units (ICUs). Group 1 included patients from June 2003 to May 2005 before the implementation of MDRs, and Group 2 included patients after the institution of MDRs from June 2005 to May 2007. In Group 1, there were 83 ventilator-associated pneumonias (VAPs) during 2414 ventilator days. In Group 2, there were 49 VAPs during 2094 ventilator days. The ratio of VAPs per thousand ventilator days decreased from 34.4 to 23.4 between the two groups (P = 0.04). When comparing trauma patients in our open ICU with similar mean Injury Severity Score and mean Abbreviated Injury Score for chest and for head and neck, implementing MDRs significantly decreased our incidence of VAP. PMID- 19999907 TI - The use of barbiturate coma as salvage therapy in a postoperative Jehovah's Witness patient with life-threatening anemia. AB - Management of the patient who refuses blood products based on religious grounds poses ethical and clinical challenges, especially when the degree of anemia becomes life-threatening. In this report, we present the case of a 52-year-old Jehovah's Witness with sickle cell disease in whom he and his family refused blood products for the treatment of severe anemia associated with profound and progressive acidosis, acute oliguric renal failure, and hemodynamic instability. Attempts carried out during the first 3 hospital days to stabilize the patient using standard therapies to support oxygen delivery as well as the use of sedation, pain control, temperature control, neuromuscular blockade, and mechanical ventilation to reduce oxygen demand were not successful. Thus, because oxygen consumption by the brain represents approximately 20 per cent of the body's oxygen needs, and pentobarbital's primary action is as a central nervous system depressant, the induction of pentobarbital coma was instituted to reduce cerebral oxygen consumption. The institution of pentobarbital on hospital Day 3 was sufficient to acutely stabilize the patient's deteriorating metabolic state and ultimately was associated with survival. Thus, we conclude that there is a potential role for barbiturate coma in Jehovah's Witness patients who refuse blood transfusions and dying of anemia when other modalities of support are not sufficient. PMID- 19999908 TI - Volvulus of the small bowel in adults. AB - The volvulus of the small bowel is a surgical emergency, causing small bowel obstruction. We performed a retrospective study of all the patients diagnosed and treated with small bowel volvulus between 1977 and 2007 at our institution. One hundred twenty-nine patients were analyzed. Thirty-nine patients presented primary volvulus and 90 secondary ones. The most frequent symptom was sudden abdominal pain. CT scan was the best diagnostic method with an accuracy of 83 per cent. Necrotic small bowel loops appeared in 46.5 per cent of the patients. Eighteen patients had postoperative complications (14%). Mortality rate was 9.3 per cent. A higher mortality is observed among patients with previous abdominal surgeries and cardiopathies. Necrotic loops are associated with higher mortality and incidence of surgical complications; patients with diabetes are associated with a higher incidence of necrotic loops. Cardiopathies are associated with more frequent medical and surgical complications. Recurrence rate was 3.9 per cent associated with simple devolvulation. Primary volvulus are more frequent among males and patients with diabetes. Jejunal location is associated with primary volvulus and these correlate with a higher incidence of necrotic loops. Primary volvulus presents a higher incidence of surgical complications. A bowel obstruction with sudden abdominal pain must be suspicious of small bowel volvulus. The main aim is to achieve an early diagnosis to prevent a necrotic small bowel. CT scan is the imaging test with the best diagnostic accuracy. Primary volvulus, the presence of necrotic loops, and patients with cardiopathies, diabetes mellitus, and with previous abdominal surgery are associated with a worse outcome. PMID- 19999909 TI - Apoptosis and microvessel density in gastric cancer: correlation with tumor stage and prognosis. AB - Gastric cancer remains one of the most common human malignancies with a poor prognosis. Apoptosis is known to be a programmed cell death and its inhibition is involved in the unregulated cellular growth that leads to neoplasms. Microvessel density (MVD) has been investigated as a promoting factor for angiogenesis with conflicting results about its relation to survival. The aim of our study was to search a correlation between these factors and some clinicopathological features and prognosis. Identification of apoptotic cells was performed applying the terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick-end labeling technique and recorded as apoptotic index (A.I.), whereas monoclonal antibodies were used for the study of MVD. A significant correlation was found between low and high A.I. and the subgroup of patients in Stages I and II (P < 0.02); 20 per cent of patients with a low A.I. showed an overall survival longer than 5 years versus 44 per cent of patients with an high A.I. (P = 0.041). High MVD was significantly related to the T stage (P = 0.036) and to a poorer 5-year overall survival (P < 0.05). Further studies are required to confirm the role of apoptosis and MVD in the development and progression of gastric cancer. PMID- 19999910 TI - Laparoscopic repair of a man with massive bilateral femoral hernias. AB - In this review, we describe a laparoscopic totally extraperitoneal (TEP) repair of a man with massive bilateral femoral hernias that had been chronically incarcerated, which has not previously been described in the literature. Our purpose is to not only to describe our laparoscopic technique and postoperative management, but also to comprehensively review the literature regarding the principles of laparoscopic hernia repair and specifically laparoscopic femoral hernia repair. TEP repair is a safe approach to the surgical management of femoral hernias, including those that are incarcerated, and furthermore offers the advantage of repairing other concomitant hernias, which may be more prevalent than expected. PMID- 19999911 TI - Recurrent abdominal compartment syndrome: an inciting factor of the second hit phenomenon. AB - Intra-abdominal hypertension (IAH) after damage control laparotomy (DCL) is not unusual and because of this, patients are treated with open-abdomen techniques to prevent abdominal compartment syndrome (ACS). The occurrence of recurrent ACS (R ACS) after abdominal wall closure under tension in patients managed with DCL can be a trigger factor for second hit syndrome. Outcomes in this subset have not been previously described. In this 1-year retrospective study of severely injured patients in a Level I trauma center managed with DCL and sequential abdominal wall closure, 26 patients were identified. After attempted abdominal wall closure, 13 (50%) patients had R-ACS and 13 (50%) non-R-ACS. R-ACS patients had a statistically significant higher incidence of multisystem organ failure, acute respiratory distress syndrome, and sepsis as well as requiring longer ventilator support and longer hospital length of stay. We concluded that failure to recognize and treat IAH with development of R-ACS after tension abdominal wall closure in patients with DCL will trigger the second hit syndrome with increased risk of morbidity. Institution of a management algorithm with intra-abdominal pressure/abdominal perfusion pressure surveillance at the time of abdominal wall closure can potentially ameliorate complications. PMID- 19999912 TI - The management of terminal ileum diverticulitis. AB - Terminal ileum diverticulitis is a very rare disease that may lead to an acute abdomen mimicking appendicitis. Because of its rarity, an optimal treatment for this disease has not been established yet. In this study, we evaluated the clinical features and outcomes of nonoperative management of terminal ileum diverticulitis, including suspected perforation. From 2000 to 2007, 346 patients were treated for acute symptomatic right-sided diverticulitis. Radiographic evaluation revealed that nine patients (2.6%) had terminal ileum diverticulitis. All patients presented with pain in the right lower quadrant. The clinical features and outcomes, including recurrence, were evaluated from a collected database and by phone interview. The average age of the patients was 43.7 years. Radiographic imaging was used to diagnose diverticulitis in all patients. Two of nine patients had suspected perforated diverticulitis, and three patients had multiple diverticula. Nonoperative management was successfully carried out on these patients. The average hospital stay was 8 days and the duration of antibiotic treatment was 6.4 days. The median follow-up was 36 months, and patients showed no recurrence within this interval. Nonoperative management may be feasible to treat terminal ileum diverticulitis. PMID- 19999913 TI - Clostridium difficile enteritis after colectomy. AB - Clostridium difficile infection of the colon is, unfortunately, a relatively common occurrence that typically follows treatment with antibiotics; however, C. difficile infection of the small bowel is a much more rare phenomenon with only 19 cases reported to date. We present three cases of isolated C. difficile enteritis after colectomy. Although all three patients were identified early and successfully treated with medical management without the need for surgical intervention, previous authors have suggested a much higher morbidity and mortality rate with this infection. This article reviews the current available literature on C. difficile enteritis to highlight this potentially serious condition in postoperative colectomy patients who present with low-grade fevers, abdominal or pelvic pain, and increased ileostomy output. PMID- 19999914 TI - Vasopressin in hemorrhagic shock: review article. AB - Trauma with resultant hypovolemic shock remains both prevalent and difficult to treat. Standard strategies using volume resuscitation and catecholamine support have historically yielded poor results. Vasopressin has emerged as a possible pharmacologic adjunct, particularly in patients with shock refractory to the administration of fluids and catecholamines. Much of the data regarding vasopressin is extrapolated from its usefulness in cases of nonhypovolemic human shock, which are supported by convincing animal studies. It is true that humans show a deficiency in vasopressin minutes after significant hemorrhage that can respond to administration of exogenous vasopressin. When given in physiological dosing regimens, vasopressin appears to be a safe adjunct to other therapy. Definite recommendations regarding indications for use, recommended dose, and long-term outcome in patients with hemorrhagic shock await a much needed prospective, randomized, controlled trial. PMID- 19999916 TI - Premonition of death in trauma: a survey of healthcare providers. AB - Prognostication in western medicine has traditionally been based on objective scientific criteria, yet providers often rely on a feeling or "sense" based on experience to provide prognoses. In trauma, some providers believe that patients who express a feeling of death are more likely to die. We randomly surveyed 302 members of the Eastern Association for the Surgery of Trauma regarding patient's premonitions of death (POD). Ninety-five per cent of respondents reported encountering patients who expressed POD. Fifty per cent agreed patients expressing POD had a higher mortality rate. Fifty-seven per cent believe patient willpower affects outcome. Forty-four per cent agreed patients have an innate ability to sense their ultimate outcome after injury; 85 per cent believe patient's POD do not cause deviations from protocols. Most trauma providers have encountered patients expressing POD. Whereas most believe that a patient's willpower affects outcome, they do not believe that expressing a desire to live decreases mortality with seemingly fatal injuries. Providers who have witnessed negative POD believe these patients are more likely to die, however, they do not deviate from treatment protocols. This survey represents the first attempt to understand the magnitude of premonition of death in trauma and the need for future research. PMID- 19999915 TI - Minor hepatic resection using heat coagulative necrosis. AB - Intra- and postoperative bleeding represents an extremely serious and frequent complication of hepatic surgery. In this study, we evaluated the effectiveness of a radiofrequency (RF) device using heat to cause coagulative necrosis of the hepatic parenchyma to control hemostasis in minor hepatic resection. Between December 2005 and November 2007, a study was conducted of 21 patients undergoing 22 hepatic resections with the RF-assisted technique. Sixteen of these were affected by hepatocellular carcinoma and five had liver metastases from colorectal cancer. Intraoperative blood loss, the need for blood transfusion, the complication rates, operating times, and the duration of postoperative hospitalization were evaluated. Four segmentectomies and 18 tumor-ectomies were performed. The average blood loss was of 15.7 mL (range, 0-40 mL); the average operating time was 25.7 minutes (range, 12-43 minutes); the mean postoperative hospital stay was 8.2 days (range, 3-49 days) with a median of 6.0 days. The authors concluded that the RF-assisted technique can be a useful method not only for reducing blood loss and avoiding blood transfusions, but also for reducing operating time and postoperative hospitalization for minor liver resections. PMID- 19999917 TI - Open-book pelvic fractures with perineal open wounds: a significant morbid combination. AB - Open-book pelvic fractures (OBPF) with concomitant intra-abdominal injuries carry a high morbidity and mortality; the significance of associated perineal open wound (OBPF-POW) has not been defined. We hypothesize that the presence of perineal open wounds increases morbidity, mortality, and concomitant use of hospital resources. Patients diagnosed with OBPF over a 5-year period at a Level I trauma center were identified by trauma registry review, and were retrospectively reviewed under an Institutional Review Board-approved protocol. Patients with OBPF without a perineal open wound were compared with those with OBPF-POW. Data collected included patient demographics, injury details, management, and outcomes. A total of 1,635 patients with blunt pelvic fractures were identified, of which 177 (10.8%) had OBPF. OBPF-POW (36/177) significantly increased the use of angioembolization, occurrence of sepsis, pelvic sepsis, ARDS, and multi-organ system failure. Patients with OBPF-POW had an increase of 13 days in length of hospitalization compared with the OBPF group (P < 0.001), with cost of $120,647.30 and $62,952.72 respectively (P < 0.001). Perineal open wounds complicate open-book pelvic fractures with significant increase in hospital resource utilization. Aggressive multidisciplinary evaluation and management is appropriate to detect and prevent complications. PMID- 19999919 TI - Anatomical and surgical considerations on lumbar hernias. AB - Lumbar hernias, which are rare hernias of the posterolateral abdominal wall, can be divided into two groups: primary lumbar hernias, often the expression of a congenital defect, which typically arise in two areas of weakness, the superior triangle and inferior triangle and acquired (or diffuse) lumbar hernias which are usually due to previous lumbar trauma or surgery. Clinical examination may be adjuvated by ultrasound or CT scan, which can reveal the abdominal wall defect with the hernia content (viscera or extraperitoneal tissue). Surgical repair of lumbar hernias, both primary and acquired, has rapidly developed through recent years, similarly to the treatment of more frequent kinds of hernia (groin, epigastric), evolving from direct repair to mini-invasive techniques, even if, since the rarity of these hernias, precise knowledge of this complex anatomic region is required. Nowadays there are two valid alternatives: open tension-free repair (with use of mesh), and mini-invasive repair. Both are safe and effective, even if smaller hernias can be treated by open approach, with loco-regional anesthesia and good cosmetic effect. Larger hernias, or hernias with suspected viscera involvement, should require larger incisions and viscera exploration. For this reason laparoscopic access would be preferable. PMID- 19999918 TI - The effect of ACGME duty hour restrictions on operative continuity of care. AB - Resident work restrictions limit participation in operations that address problems created by a prior operation, because complications occur at any time. We compared resident and attending surgeon staffing of operative complications. We reviewed all complications that required a second operation reported at our Morbidity and Mortality Conference over 1 year, noting surgeons present, their postgraduate year level, and call shift. Comparisons were done using chi2. Of 142 cases, 39 involved a second operation. The same attending surgeon was present for both in 79 per cent of cases, whereas the same resident was present in only 44 per cent (P = 0.002). Postgraduate year 4 to 5 were less likely to be present for second operations than attendings (48% vs 87%, P = 0.011). Resident shift (day, night float, and weekend) was known in 32 cases. When the first operation occurred during day hours, attendings and residents were equally likely to be present at the second (55% and 45%, P = 0.16). When original operations took place during night float or weekend shifts, residents were less likely to be present (33%) than attendings (83%) at second operations (P = 0.036). Duty hour restrictions interfere with operative continuity of care. Reoperations should be exempted from duty hour restrictions. PMID- 19999920 TI - A betrayal of our handiwork: postoperative hemorrhage and the need for reoperation. AB - Postoperative hemorrhage (PH) that requires reoperation to control bleeding represents a potentially life-threatening and avoidable complication that could have serious implications for recovery. All surgical patients were reviewed who developed PH and required reoperation for control of hemorrhage over a 12-year period, to examine contributing factors possibly related to surgeon misadventure. Of 89,663 operations during this period, there were 1,031 patients (1.2%) who developed PH. Of these, 36 patients required reoperation for control of PH (0.04%), including, general surgery (17), otolaryngologic (9), cardiovascular (9), and gynecologic (1) patients. In 27 general, cardiovascular, and gynecologic patients (29 reoperations), the age ranged from 6 to 91 years. Almost one-half of patients (56%) developing PH were on preoperative anticoagulation. Estimated operative blood loss (EBL) was moderate (EBL = 100-500 mL, 48%). Most patients were normothermic (80%) and normotensive (93%) intraoperatively. The decision to reoperate was not made for at least 8 hours in 55 per cent of patients. At reoperation 10/29 patients were hypotensive. In 20/36 patients (56%) the reoperation note did not identify a single source of bleeding. PH is a distinctly uncommon complication of surgery and often not due to obvious surgeon misadventure. Reoperation for PH is even rarer and embarked upon with reluctance, frequently not yielding a discernible cause for hemorrhage. PMID- 19999921 TI - Fast track postoperative management after elective colorectal surgery: a controlled trail. AB - In the attempt to reduce postoperative complications and costs and improve outcomes, the concept of fast track surgery has been proposed. Improvements in anesthesia techniques and a better understanding of the pathophysiologic events occurring during and after surgery have made it possible. A group of patients undergoing colorectal resections with a fast track approach were investigated; specifically, the effects on postoperative morbidity, resumption of intestinal function, and duration of hospitalization. Fifty patients were managed according to a protocol, which included epidural analgesia, early ambulation, and oral feeding (fast track group); they were compared with 50 patients managed with a different protocol: no epidural analgesia, early ambulation, and early oral diet (control group). Primary outcome end-points reported include morbidity, time to passage of flatus and stool, and length of hospital stay. Fourteen complications occurred in the fast track group and 13 in the control group (P = not significant (NS)). Resumption of intestinal function occurred after 3 days, and length of hospital stay was 5 days in the fast track group compared with 4 and 7 days respectively in control patients (P = NS, P < 0.01). Patients undergoing elective colorectal resections can be managed safely with fast track protocols reducing hospital stay. PMID- 19999922 TI - Internal herniation of the cecum through the foramen of Winslow. PMID- 19999923 TI - Successful treatment of ascending aortic graft and valve endocarditis after debulking splenectomy for invasive aspergillosis and mucor. PMID- 19999924 TI - Extraosseous Ewing's sarcoma. PMID- 19999925 TI - Novel minimally invasive approach in the management of body packers. PMID- 19999926 TI - Extracorporeal membrane oxygenation for severe hypoxemia after trauma pneumonectomy. PMID- 19999927 TI - Compartment syndrome of the leg after pelvic surgery: can it always be avoided? PMID- 19999928 TI - Poorly differentiated thyroid cancer metastasis to the sternum. PMID- 19999929 TI - Bowel obstruction secondary to internal hernia after laparoscopic Nissen fundoplication and gastrostomy tube placement. PMID- 19999930 TI - Methicillin-resistant Staphylococcus aureus pulmonary valve endocarditis. PMID- 19999931 TI - A pseudoaneurysm of the inferior epigastric artery after laparoscopic right hemicolectomy. PMID- 19999932 TI - Successful salvage of right common iliac vein injury in a 10-year-old boy undergoing laparoscopic appendectomy. PMID- 19999933 TI - Be proactive: changing the scenario for nurse graduates. PMID- 19999934 TI - Nursing shortage solutions and America's economic recovery. PMID- 19999935 TI - Clinical evaluation and grading practices in schools of nursing: national survey findings part II. AB - To better understand how nurse educators evaluate and grade students' clinical practice, the Evaluation of Learning Advisory Council of the National League for Nursing conducted a survey of faculty (N = 1,573) in all types of prelicensure RN programs. This article describes the findings of that survey in relation to clinical evaluation and grading clinical practice. Nearly all faculty used a clinical evaluation tool to rate students' performance in the clinical setting (n = 1,534, 98 percent); most programs had the same basic tool in all courses, but modified to reflect the unique aspects of each course (n = 1,095, 70 percent). Faculty (n = 1,116, 83 percent) reported using pass/fail for grading in clinical courses rather than a letter or numerical grade. PMID- 19999936 TI - Internalizing the concept of caring: an examination of student perceptions when nurses share their stories. AB - Caring should be incorporated in the curriculum and the focus of nursing education. However, measurement of nurses' perceptions of caring abilities remains an area that is not well explored. This pilot study explored nursing students' perceptions of caring and their ability to form caring relationships with clients after hearing clinical nurses discuss their own caring experiences. Fourteen nursing students met with three clinical nurses who shared their stories about caring practice. Results indicate that although there was no statistically significant change, the mean score for caring efficacy was increased postintervention. Student comments also reflect that sharing stories about caring was beneficial, valued, and potentially an effective teaching strategy to help students internalize the concept of caring. PMID- 19999937 TI - Doctor of Nursing Practice program evaluation and beyond: capturing the profession's transition to the DNP. AB - The Doctor of Nursing Practice (DNP) educational preparation for advanced practice nursing and DNP role development are innovations that require intense scrutiny through formative and summative evaluation. It is imperative to move beyond standard curriculum evaluation approaches to capture the transformation of advanced nursing practice and the profession. The University of Washington School of Nursing designed a comprehensive DNP evaluation plan that focuses on both program effectiveness and the experience of transition as described by faculty and students. The evaluation plan is outlined, and lessons learned regarding data collection, student and faculty participation, and institutional review board approval are discussed. Examples of instruments are included. PMID- 19999938 TI - Expert to novice: clinicians learning new roles as clinical nurse educators. AB - The nursing shortage presents a challenge for both nursing education programs and clinical agencies to identify creative options for increasing the supply of nurse educators, especially those who can teach vital clinical skills to future nurses. Some clinicians have voiced that they would enjoy having opportunities to share their clinical expertise through teaching, but they lack preparation for this new role. Through written narratives of nurses enrolled in a Clinical Nurse Educator Academy, this study explored the research question: How can narratives of clinicians participating in a Clinical Nurse Educator Academy enhance understanding of the unique perspectives of these individuals as they prepare for roles as clinical nurse educators? Data consisted of 135 reflective narratives from 45 participants and were analyzed using van Manen's approach to human science research. One overarching pattern, "The Phenomenon of Learning to Teach:" and three themes,"Buckle Your Seatbelt," "Embracing the Novice," and "Mentoring in the Dark," emerged from the data. Implications for faculty and further research are offered. PMID- 19999939 TI - A model of preceptorship in nursing: reflecting the complex functions of the role. AB - Preceptorship has been consistently acknowledged in the literature as a strategy to maximize the benefits of clinical nursing education in terms of knowledge and skill acquisition, confidence, and professional socialization. Further benefits have also been recognized for preceptors, and for the broader health care agency. Despite recognition of the importance of this role, there has been no clearly articulated model of preceptorship that reflects the broader factors impacting upon the relationship between registered nurse and nursing student. This article presents a model of preceptorship that is directly based upon the preceptor preceptee relationship and reflects the factors and influences that might impact, both positively and negatively, the strength and effectiveness of the relationship and subsequent learning outcomes. This model will provide a useful resource for the planning and implementation of a preceptorship program that maximizes clinical learning to the satisfaction of all stakeholders. PMID- 19999940 TI - A new model for the clinical instruction of undergraduate nursing students. AB - As the nursing faculty shortage worsens nationwide, schools of nursing must be creative in developing models of clinical instruction for undergraduate students that ensure both quality instruction and quality patient care. Partnerships with clinical agencies can be creatively designed to allow full-time faculty greater access to students and agency nurses recognition for clinical expertise. The model for clinical instruction proposed here is also useful for introducing staff nurses to the role of faculty. The program described here was piloted for one semester; proposed advantages and disadvantages are described, and feedback from students is presented. PMID- 19999941 TI - Collaborative teaching in a virtual environment to promote conceptual change for nurse educator master's students. AB - A virtual course for the nurse educator master's program was developed and delivered collaboratively between two professors, one from the College of Health Professions (CHP) and the other from the College of Liberal Arts and Education (CLAE). The collaborative effort provided a venue for both professors to complement each other's strengths, affording students the benefit of constructivist pedagogy using content appropriate for a nurse educator program. Student discussion board communication indicated the anticipated conceptual change as a result of teaching using constructivist methodology. Qualitative data were collected and will be used to improve the course as well as develop the follow-up course. PMID- 19999942 TI - From beta-blockers to boot camp: Preparing students for the NCLEX-RN. AB - Although it is thought that reviewing essential materials and learning how to answer computer-generated questions are optimal preparations for NCLEX-RN, strategies that build knowledge, self-confidence, and professionalism of the nurse taking the exam are equally important. A senior seminar course that guides formal NCLEX-RN preparation is presented in this article with specific course strategies and a blueprint of seminar content that can be adapted to the nursing curriculum. PMID- 19999943 TI - The usefulness of personal digital assistants (PDAs) to nursing students in the clinical setting: a pilot study. PMID- 19999944 TI - Digital object identifiers: are they important to me? PMID- 19999945 TI - Reinvigorating a key research program: the NLN's collection of LPN/LVN educational statistics. PMID- 19999946 TI - A letter to Sarah. PMID- 19999947 TI - The 3rd International Cancer Control Congress: international collaboration in an era of cancer as a global concern. PMID- 19999948 TI - Cancer control-planning and monitoring population-based systems. AB - Cancer is a growing global health issue, and many countries are ill-prepared to deal with their current cancer burden let alone the increased burden looming on the horizon. Growing and aging populations are projected to result in dramatic increases in cancer cases and cancer deaths particularly in low- and middle income countries. It is imperative that planning begin now to deal not only with those cancers already occurring but also with the larger numbers expected in the future. Unfortunately, such planning is hampered, because the magnitude of the burden of cancer in many countries is poorly understood owing to lack of surveillance and monitoring systems for cancer risk factors and for the documentation of cancer incidence, survival and mortality. Moreover, the human resources needed to fight cancer effectively are often limited or lacking. Cancer diagnosis and cancer care services are also inadequate in low- and middle-income countries. Late-stage presentation of cancers is very common in these settings resulting in less potential for cure and more need for symptom management. Palliative care services are grossly inadequate in low- and middle-income countries, and many cancer patients die unnecessarily painful deaths. Many of the challenges faced by low- and middle-income countries have been at least partially addressed by higher income countries. Experiences from around the world are reviewed to highlight the issues and showcase some possible solutions. PMID- 19999950 TI - Cancer prevention and population-based screening. AB - Cancer prevention, screening and early detection can provide some of the greatest public health benefits for cancer control. In low resource settings, where cancer control is challenged by limited human, financial and technical resources, cancer prevention and screening are of utmost importance and can provide significant impacts on the cancer burden. Public policies, social, environmental and individual level interventions which promote and support healthy eating and physical activity can lower cancer risks. Tobacco use, a significant cancer risk factor, can be reduced through the application of key mandates of the World Health Organization Framework Convention on Tobacco Control. In addition, cancer screening programs, namely for cervical and breast cancers, can have a significant impact on reducing cancer mortality, including in low resource settings. Comprehensive cancer control programs require interventions for cancer prevention, screening and early detection, and involve sectors outside of health to create supportive environments for healthy ways of life. Sharing experiences in implementing cancer control programs in different settings can create opportunities for interchanging ideas and forming international alliances. PMID- 19999949 TI - International collaborations in cancer control and the Third International Cancer Control Congress. AB - Over the past few decades, there has been growing support for the idea that cancer needs an interdisciplinary approach. Therefore, the international cancer community has developed several strategies as outlined in the WHO non communicable diseases Action Plan (which includes cancer control) as the World Health Assembly and the UICC World Cancer Declaration, which both include primary prevention, early diagnosis, treatment, and palliative care. This paper highlights experiences/ideas in cancer control for international collaborations between low, middle, and high income countries, including collaborations between the European Union (EU) and African Union (AU) Member States, the Latin-American and Caribbean countries, and the Eastern Mediterranean countries. These proposals are presented within the context of the global vision on cancer control set forth by WHO in partnership with the International Union Against Cancer (UICC), in addition to issues that should be considered for collaborations at the global level: cancer survival (similar to the project CONCORD), cancer control for youth and adaptation of Clinical Practice Guidelines. Since cancer control is given lower priority on the health agenda of low and middle income countries and is less represented in global health efforts in those countries, EU and AU cancer stakeholders are working to put cancer control on the agenda of the EU-AU treaty for collaborations, and are proposing to consider palliative care, population based cancer registration, and training and education focusing on primary prevention as core tools. A Community of Practice, such as the Third International Cancer Control Congress (ICCC-3), is an ideal place to share new proposals, learn from other experiences, and formulate new ideas. The aim of the ICCC-3 is to foster new international collaborations to promote cancer control actions in low and middle income countries. The development of supranational collaborations has been hindered by the fact that cancer control is not part of the objectives of the Millennium Development Goals (MGGs). As a consequence, less resources of development aids are allocated to control NCDs including cancer. PMID- 19999951 TI - Comprehensive cancer control-research & development: knowing what we do and doing what we know. AB - Comprehensive cancer control is defined as an integrated and coordinated approach to reducing cancer incidence, morbidity, and mortality across the cancer control continuum from primary prevention to end-of-life care. This approach assumes that when the public sector, non-governmental organizations, academia, and the private sector share with each other their skills, knowledge, and resources, a country can take advantage of all its talents and resources to more quickly reduce the burden of cancer for all its population. One critical issue for comprehensive cancer control is the extent to which the private sector can contribute to cancer prevention and control programs and policies that have historically been lead by the public health sector, and similarly how can the public sector increase its investment and involvement in clinical research and practice issues that are largely driven by the private sector worldwide? In addition, building capacity to integrate research that is appropriate to the culture and context of the population will be important in different settings, in particular research related to cancer control interventions that have the capacity to influence outcomes. To whatever extent cancer control research is ultimately funded through the private and public sectors, if investments in research discoveries are ultimately to benefit the populations that bear the greatest burden of disease, then new approaches to integrating the lessons learned from science with the lessons learned from service (public health, clinical, and public policy) must be found to close the gap between what we know and what we do. Communities of practice for international cancer control, like the ones fostered by the first three International Cancer Control Congresses, represent an important forum for knowledge exchange opportunities to accelerate the translation of new knowledge into action to reduce the burden of cancer worldwide. PMID- 19999952 TI - Organization of population-based cancer control programs: Europe and the world. AB - As cancer is to a large extent avoidable and treatable, a cancer control program should be able to reduce mortality and morbidity and improve the quality of life of cancer patients and their families. However, the extent to which the goals of a cancer control program can be achieved will depend on the resource constraints a country faces. Such population-based cancer control plans should prioritize effective interventions and programs that are beneficial to the largest part of the population, and should include activities devoted to prevention, screening and early detection, treatment, palliation and end-of-life care, and rehabilitation. In order to develop a successful cancer control program, leadership and the relevant stakeholders, including patient organizations, need to be identified early on in the process so that all partners can take ownership and responsibility for the program. Various tools have been developed to aid them in the planning and implementation process. However, countries developing a national cancer control program would benefit from a discussion of different models for planning and delivery of population-based cancer control in settings with differing levels of resource commitment, in order to determine how best to proceed given their current level of commitment, political engagement and resources. As the priority assigned to different components of cancer control will differ depending on available resources and the burden and pattern of cancer, it is important to consider the relative roles of prevention, early detection, diagnosis, treatment, rehabilitation and palliative care in a cancer control program, as well as how to align available resources to meet prioritized needs. Experiences from countries with differing levels of resources are presented and serve to illustrate the difficulties in developing and implementing cancer control programs, as well as the innovative strategies that are being used to maximize available resources and enhance the quality of care provided to cancer patients around the world. PMID- 19999953 TI - Critical factors influencing the establishment, maintenance and sustainability of population-based cancer control programs. AB - Developing and maintaining a comprehensive cancer control program are two distinct entities. Key issues related to building and sustaining cancer control programs include how to integrate initiatives and efforts across multiple constituencies addressing components of the implementation of cancer control and non-communicable disease programs, the processes used in different resource settings to achieve effective drug budgeting, health technology assessment and health economics, and how countries can support public and societal engagement. There are promising examples in both resource-rich and resource-challenged countries of constituencies that have developed programs which can contribute to comprehensive cancer control. Some take advantage of newer technology and information services, while others are more people and patient focused. Critical issues and factors for establishing and maintaining population-based comprehensive cancer control programs are identified and reviewed. PMID- 19999954 TI - Respiratory home care: take the pulmonary specialist out of the hospital! PMID- 19999955 TI - Estimates of PI*S and PI*Z Alpha-1 antitrypsin deficiency alleles prevalence in the Caribbean and North, Central and South America. AB - BACKGROUND: AAT deficiency is not a rare disease, but one of the most common congenital disorders increasing susceptibility of individuals with this deficiency to both lung and liver disease as well as other several adverse health effects. Studies to develop accurate estimates of the magnitude of this genetic disorder in any given country is critical for the development of screening programs for detection, diagnosis, and treatment of those individuals and/or families at risk. In the present study, estimates of the prevalence of the two major deficiency alleles PI S and PI Z were estimated for 25 countries in the Caribbean and North, Central, and South America to supplement our previous studies on 69 countries worldwide. METHOD: Using data on the prevalence of the two most common deficiency alleles PI S and PIZ in the mother countries that provided the majority of immigrants to these 25 countries, as well as genetic epidemiological studies on various genetic subgroups indigenous to the Caribbean and North, Central and South America it was possible to develop new formulas to estimate the numbers in each of five phenotypic classes, namely PI MS, PI MZ, PI SS, PI SZ and PI ZZ for each country. RESULTS: When these 25 countries were grouped into six different geographic regions, the present study demonstrated striking differences when comparisons were made in numeric tables, maps and figures. Highly significant numbers of individuals at risk for AAT Deficiency were found in both the European, Mestizo and Mulatto populations for most of the 25 countries studied in the Caribbean and North, Central and South America. CONCLUSIONS: Our studies demonstrated striking differences in the prevalence of both the PIS and PIZ alleles among these 25 countries in the Caribbean and North, Central and South America and significant numbers of individuals at risk for adverse health effects associated with AAT Deficiency in a given country. When these data are added to the results from our earlier studies on 69 countries, we now have data on AAT Deficiency in 94 of the 193 countries worldwide listed in the CIA FactBook. PMID- 19999956 TI - Comprehensive exercise training improves ventilatory muscle function and reduces dyspnea perception in patients with COPD. AB - BACKGROUND: Comprehensive exercise training (CET) is an efficient strategy to decrease dyspnea perception in chronic obstructive pulmonary disease (COPD) and may result in significant improvement in ventilatory muscles function. Our aim was to evaluate the effects of general exercise training on dyspnea perception and on respiratory muscles strength in COPD patients. METHODS: Consecutive COPD patients were enrolled to complete a CET programme. The patients underwent a routine that included a global warm up, upper and lower limbs endurance exercise as well as stretching and relaxation. Before and after the CET programme, patients completed maximal inspiratory (PImax) and expiratory (PEmax) pressures measurements, maximal incremental test, endurance test, and 6-min walk distance (6MWD). RESULTS: 71 patients (52 male). Mean age 67.6 +/- 8.6 years, FEV1 (%) 44.2 +/- 16.2 and Mahler dyspnea scale 6.4 +/- 1.8. The results before and after the exercise programme were: PImax 64.7 +/- 22.9 vs. 75.5 +/- 23.7 cmH2O (p=0.001), PEmax 110.8 +/- 28.1 vs. 120.4 +/- 28.1 cmH2O (p=0.004), 6MWD 510.6 +/ 90.3 vs. 528.2 +/- 99.7 metres (p=0.88), time of incremental test 672 +/- 135 vs. 856 +/- 226 sec (p<0.0001). Compared with the pre exercise programme, we observed a significant reduction on Borg dyspnea scale (6.1 +/- 2.8 to 3.6 +/- 2.3, p<0.0001) as well as a longer test time (504 +/- 218 to 1.038 +/- 841, p<0.0001) at the end of the endurance test after CET programme. Improvement of PImax correlated negatively with dyspnea perception at iso-time during the endurance test (r = -0.33, p=0.03). CONCLUSIONS: Our results confirm that CET is associated with significant improvement in PImax, PEmax and provide evidence demonstrating that CET reduces dyspnea perception in patients with COPD. PMID- 19999957 TI - Episodic medical home interventions in severe bedridden chronic respiratory failure patients: a 4 year retrospective study. AB - BACKGROUND AND AIM: Home care for respiratory patients includes a complex array of services delivered in an uncontrolled setting. The role of a respiratory specialist inside the home healthcare team has been scarcely studied up to now. Our aims were to analyse the number and quality of episodic home visits performed by respiratory physicians to severe bedridden Chronic Respiratory Failure (CRF) patients, and also to evaluate the safety of tracheotomy tube substitutions at home. METHODS: 231 home interventions (59.8/year) in 123 CRF patients (59 males; age 63 +/- 17 y, 24 on oxygen therapy, 35 under non invasive mechanical ventilation, 46 under invasive ventilation, 74 with tracheostomy) located 35 +/- 16 km far from referred hospital, were revised in a period of 4 years (2005 2008). RESULTS: Chronic Obstructive Pulmonary Disease (COPD) (31%) and amyotrophic lateral sclerosis (ALS) (28%) were the more frequent diagnoses. Interventions were: tracheotomy tube substitution (64%) presenting 22% of minor adverse events and 1.4% of major adverse events; change or new oxygen prescription (37%); nocturnal pulsed saturimetric trend prescription (24%); change in mechanical ventilation (MV) setting (4%); new MV adaptation (7%). After medical intervention, new home medical equipment devices (oxygen and MV) were prescribed in 36% of the cases while rehabilitative hospital admission and home respiratory physiotherapy prescription was proposed in 9% and 6% of the cases respectively. Patient/caregiver's satisfaction was reported on average 8.48 +/- 0.79 (1 = the worst; 10 = the higher). The local health care system (HCS) reimbursed 70 euros for each home intervention. Families saved 42 +/- 20 euros per visit for ambulance transportation. CONCLUSIONS: Home visits performed by a respiratory physician to bedridden patients with chronic respiratory failure: 1. include predominantly patients affected by COPD and ALS; 2. determine a very good satisfaction to patients/caregivers; 3. allow money saving to caregivers; 4. are predominantly made up to change tracheotomy tube without severe adverse events. PMID- 19999958 TI - Markers of airway inflammation in pulmonary diseases assessed by induced sputum. AB - During recent years there has been a growing interest in using non-invasive biomarkers to understand and monitor the airway inflammation in subjects with respiratory tract disorders. To date, the best validated and performing non invasive biomarkers are measures of inflammation in induced sputum in both cellular and fluid phase, which can provide biological insights into the pathogenesis of respiratory diseases such as asthma and chronic obstructive pulmonary disease. The purpose of this review is to examine the principal literature on the different markers of inflammation in pulmonary diseases assessed by induced sputum analysis in either cellular or fluid phase. PMID- 19999959 TI - Time to culture conversion in smokers with pulmonary tuberculosis. AB - BACKGROUND AND OBJECTIVE: It has been reported that tobacco smoking slows the sterilisation of sputum culture in pulmonary tuberculosis, but the factors that could delay culture conversion in patients who smoke are not known. Our aim is to identify the factors influencing sputum culture conversion in smokers with pulmonary tuberculosis. METHODS: Ninety-nine patients with a smoking history and diagnosed with pulmonary tuberculosis were analysed retrospectively. The relationship between sputum culture status at the second month and the following variables: age, gender, pack-years index, comorbid diseases, number acid-fast bacilli (AFB) in sputum smear examination, radiological findings (cavitary, extensive or limited disease), drug susceptibility pattern and initial treatment, was analysed. The Student t-test, chi-square test and logistic regression model with forward stepwise conditional methods were used for statistical analysis. A p value of <0.05 was considered to be statistically significant. RESULTS: Twenty six patients (26.2%): 18 males (22.2%) and 8 females (44%) were sputum culture positive at the end of the second month of treatment. In univariate analysis, culture conversion time was significantly associated with female gender and extensive disease, but in a logistic regression analysis was only correlated with female gender (OR=5.63 95% CI 1.21-20.64-p=0.02). CONCLUSION: In current smokers with pulmonary tuberculosis, the 'time to culture' conversion relates only to the female gender. PMID- 19999960 TI - Unexpected delayed death after manual strangulation: need for careful examination in the emergency room. AB - The aim of this case report was to demonstrate the importance of detailed clinical assessment in victims of attempted manual strangulation, to prevent the occurrence of delayed death due to an airway collapse; and to provide an update on clinical management of these patients. An elderly male patient presented with a sore throat and speech impairment after attempting manual strangulation. Physical examination showed reddish skin of the neck, an extensive haematoma of the hard palate and anterior tongue. Flexible laryngoscopy failed to show any swelling of the hypopharynx or larynx. A few hours after presentation, the patient developed acute dyspnoea and died. Autopsy and post-mortem CT scan showed a haematoma in the thyro-epiglottal space. In conclusion, victims of manual strangulation can survive despite internal neck injury which can lead to delayed fatal airway collapse. This is because often there are few or no signs of assault, therefore medical evaluation should be thorough and timely. Sometimes immediate tracheotomy can be life saving. PMID- 19999961 TI - Denitrifying phosphorus removal in a step-feed CAST with alternating anoxic-oxic operational strategy. AB - A bench-scale cyclic activated sludge technology (CAST) was operated to study the biological phosphorus removal performance and a series of batch tests was carried out to demonstrate the accumulation of denitrifying polyphosphate-accumulating organisms (DNPAOs) in CAST system. Under all operating conditions, step-feed CAST with enough carbon sources in influent had the highest nitrogen and phosphorus removal efficiency as well as good sludge settling performance. The average removal rate of COD, NH4+-N, PO4(3-)-P and total nitrogen (TN) was 88.2%, 98.7%, 97.5% and 92.1%, respectively. The average sludge volume index (SVI) was 133 mL/g. The optimum anaerobic/aerobic/anoxic (AOA) conditions for the cultivation of DNPAOs could be achieved by alternating anoxic/oxic operational strategy, thus a significant denitrifying phosphorus removal occurred in step-feed CAST. The denitrification of NOx(-)-N completed quickly due to step-feed operation and enough carbon sources, which could enhance phosphorus release and further phosphorus uptake capability of the system. Batch tests also proved that polyphosphate-accumulating organisms (PAOs) in the step-feed process had strong denitrifying phosphorus removal capacity. Both nitrate and nitrite could be used as electron acceptors in denitrifying phosphorus removal. Low COD supply with step-feed operation strategy would favor DNPAOs accumulation. PMID- 19999962 TI - A novel advanced oxidation process to degrade organic pollutants in wastewater: microwave-activated persulfate oxidation. AB - This article, for the first time, provides a novel advanced oxidation process based on sulfate radical (SO4*-) to degrade organic pollutants in wastewater: microwave (MW)-activated persulfate oxidation (APO) with or without active carbon (AC). Azo dye acid Orange 7 (AO7) is used as a model compound to investigate the high reactivity of MW-APO. It is found that AO7 (up to 1000 mg/L) is completely decolorized within 5-7 min under an 800 W MW furnace assisted-APO. In the presence of chloride ion (up to 0.50 mol/L), the decolorization is still 100% completed, though delayed for about 1-2 min. Experiments are made to examine the enhancement by AC. It is exciting to find that the 100% decolorization of AO7 (500 mg/L) is achieved within 3 min by MW-APO using 1.0 g/L AC as catalyst, while the degradation efficiency maintains at 50% by MW energy without persulfate after about 5 min. Besides the destruction of visible light chromophore band of AO7 (484 nm), during MW-APO, two bands in the ultraviolet region (228 nm and 310 nm) are rapidly broken down. The removal of COD is about 83%-95% for 500 mg/L AO7. SO4*- is identified with quenching studies using specific alcohols. Both SO4*- and *OH could degrade AO7, but SO4*- plays the dominant role. In a word, MW-APO AC is a new catalytic combustion technology for destruction of organic contamination even for high concentration. PMID- 19999963 TI - Feasibility and advantage of biofilm-electrode reactor for phenol degradation. AB - The new biofilm-electrode method was used for the phenol degradation, because of its low current requirement. The biofilm-electrode reactor consisted of immobilized degrading bacteria on Ti electrode as cathode and Ti/PbO2 electrode as anode. With the biofilm-electrode reactor in a divided electrolytic cell, the phenol degradation rate could achieve 100% at 18 h which was higher than using traditional methods, such as biological or electrochemical methods. Chemical oxygen demand (COD) removal rate of the biofilm-electrode reactor was also greater than that using biological and electrochemical method, and could reach 80% at 16 h. The results suggested that the biofilm-electrode reactor system can be used to treat wastewater with phenol. PMID- 19999964 TI - Total pollution effect of urban surface runoff. AB - For pollution research with regard to urban surface runoff, most sampling strategies to date have focused on differences in land usage. With single land use sampling, total surface runoff pollution effect cannot be evaluated unless every land usage spot is monitored. Through a new sampling strategy known as mixed stormwater sampling for a street community at discharge outlet adjacent to river, this study assessed the total urban surface runoff pollution effect caused by a variety of land uses and the pollutants washed off from the rain pipe system in the Futian River watershed in Shenzhen City of China. The water quality monitoring indices were COD (chemical oxygen demand), TSS (total suspend solid), TP (total phosphorus), TN (total nitrogen) and BOD (biochemical oxygen demand). The sums of total pollution loads discharged into the river for the four indices of COD, TSS, TN, and TP over all seven rainfall events were very different. The mathematical model for simulating total pollution loads was established from discharge outlet mixed stormwater sampling of total pollution loads on the basis of four parameters: rainfall intensity, total land area, impervious land area, and pervious land area. In order to treat surface runoff pollution, the values of MFF30 (mass first flush ratio) and FF30 (first 30% of runoff volume) can be considered as split-flow control criteria to obtain more effective and economical design of structural BMPs (best management practices) facilities. PMID- 19999965 TI - Kinetics and mechanisms of p-nitrophenol biodegradation by Pseudomonas aeruginosa HS-D38. AB - The kinetics and mechanisms of p-nitrophenol (PNP) biodegradation by Pseudomonas aeruginosa HS-D38 were investigated. PNP could be used by HS-D38 strain as the sole carbon, nitrogen and energy sources, and PNP was mineralized at the maximum concentration of 500 mg/L within 24 h in an mineral salt medium (MSM). The analytical results indicated that the biodegradation of PNP fit the first order kinetics model. The rate constant kPNP is 2.039 x 10(-2)/h in MSM medium, KPNP+N is 3.603 x 10(-2)/h with the addition of ammonium chloride and KPNP+C is 9.74 x 10(-3)/h with additional glucose. The addition of ammonium chloride increased the degradation of PNP. On the contrary, the addition of glucose inhibited and delayed the biodegradation of PNP. Chemical analysis results by thin-layer chromatography (TLC), UV-Vis spectroscopy and gas chromatography (GC) techniques suggested that PNP was converted to hydroquinone (HQ) and further degraded via 1,2,4-benzenetriol (1,2,4-BT) pathway. PMID- 19999966 TI - Influence of rice straw burning on the levels of polycyclic aromatic hydrocarbons in agricultural county of Taiwan. AB - Atmospheric particulate and polycyclic aromatic hydrocarbons (PAHs) size distribution were measured at Jhu-Shan (a rural site) and Sin-Gang (a town site) in central Taiwan during the rice straw burning and non-burning periods. The concentrations of total PAHs accounting for a roughly 58% (34%) increment in the concentrations of total PAHs due to rice-straw burning. Combustion-related PAHs during burning periods were 1.54-2.57 times higher than those during non-burning periods. The mass median diameter (MMD) of 0.88-1.21 microm in the particulate phase suggested that rice-straw burning generated the increase in coarse particle number. Chemical mass balance (CMB) receptor model analyses showed that the primary pollution sources at the two sites were similar. However, rice-straw burning emission was specifically identified as a significant source of PAH during burning periods at the two sites. Open burning of rice straws was estimated to contribute approximately 6.3%-24.6% to total atmospheric PAHs at the two sites. PMID- 19999967 TI - Optimization of gasoline hydrocarbon compositions for reducing exhaust emissions. AB - Effects of hydrocarbon compositions on raw exhaust emissions and combustion processes were studied on an engine test bench. The optimization of gasoline hydrocarbon composition was discussed. As olefins content increased from 10.0% to 25.0% in volume, the combustion duration was shortened by about 2 degree crank angle (degrees CA), and the engine-out THC emission was reduced by about 15%. On the other hand, as aromatics content changed from 35.0% to 45.0%, the engine-out NOx emissions increased by 4%. An increment in olefins content resulted in a slight increase in engine-out CO emission, while the aromatics content had little effect on engine-out total hydrocarbon (THC) and CO emissions. Over the new European driving cycle (NEDC), the THC, NOx and CO emissions of fuel with 25.0% olefins and 35.0% aromatics were about 45%, 21% and 19% lower than those of fuel with 10.0% olefins and 40.0% aromatics, respectively. The optimized gasoline compositions for new engines and new vehicles have low aromatics and high olefins contents. PMID- 19999968 TI - Effects of arbuscular mycorrhizal fungi inoculation on arsenic accumulation by tobacco (Nicotiana tabacum L.). AB - A pot experiment was conducted to study the effects of arbuscular mycorrhizal (AM) fungi (from contaminated or uncontaminated soils) on arsenic (As) uptake of tobacco (Nicotiana tabacum L.) in As-contaminated soil. Mycorrhizal colonization rate, dry weight, As and P uptake by plants, concentrations of water-extractable As and As fractions were determined. A low mycorrhizal colonization rate (< 25%) was detected. Our research indicated that AM fungi isolated from polluted soils were no more effective than those from unpolluted soils when grown in symbiosis with tobacco. No significant differences were observed in roots and stalks dry weights among all treatments. Leaves and total plant dry weights were much higher in Glomus versiforme treatment than that in control treatment. As contents in roots and stalks from mycorrhizal treatments were much lower than that from control treatment. Total plant As content exhibited the same trend. P concentrations in tobacco were not affected by colonization, nor were stalks, leaves and total plant P contents. Roots P contents were remarkably lower in HN treatments than in other treatments. Meanwhile, decreased soil pH and lower water extractable As concentrations and higher levels of As fraction bound to well crystallized hydrous oxides of Fe and Al were found in mycorrhizal treatments than in controls. The protective effect of mycorrhiza against plant As uptake may be associated with changes in As solubility mediated by changing soil pH. These results indicated that under As stress, proper mechanisms employed by AM fungi can protect tobacco against As uptake. Results confirmed that AM fungi can play an important role in food quality and safety. PMID- 19999969 TI - Four years of free-air CO2 enrichment enhance soil C concentrations in a Chinese wheat field. AB - Elevated atmospheric CO2 can influence soil C dynamics in agroecosystems. The effects of free-air CO2 enrichment (FACE) and N fertilization on soil organic C (Corg), dissolved organic C (DOC), microbial biomass C (Cmic) and soil basal respiration (SBR) were investigated in a Chinese wheat field after expose to elevated CO2 for four full years. The results indicated that elevated CO2 has stimulative effects on soil C concentrations regardless of N fertilization. Following the elevated CO2, the concentrations of Corg and SBR were increased at wheat jointing stage, and those of DOC and Cmic were enhanced obviously across the wheat jointing stage and the fallow period after wheat harvest. On the other hand, N fertilization did not significantly affect the content of soil C. Significant correlations were found among DOC, Cmic, and SBR in this study. PMID- 19999970 TI - Nitrification potentials of Chinese tea orchard soils and their adjacent wasteland and forest soils. AB - To investigate the nitrifying activities of different soil types, soil samples collected from 8-, 50- and 90-year old tea orchards, the adjacent wasteland, and 90-year old forest were measured for their nitrification potentials using the conventional soil incubation and the liquid incubation method. Among different soil types, the nitrification potential of soil in tea orchards was higher than that of wasteland and forest soils. The slurry shaken liquid incubation method was confirmed to be more accurate and have reliable results than the soil incubation. Interestingly, experimental result revealed that the generally applied pH value of 7.2 for the liquid media was not the optimal pH for these acid soils with a strong buffer capacity. This suggested that tea orchard soils may have nitrifiers requiring pH-neutral condition for the best activity. Our data also showed that treatment with the commonly used nitrogen fertilizer urea significantly improved nitrification potential of the soils; such enhancement effect was stronger on all of three tea orchard soils than on wasteland and forest soils, and also stronger on the younger (8- and 50-year old) tea orchard soils than on the older one (90-year old). PMID- 19999971 TI - Characterizing spatial distribution and sources of heavy metals in the soils from mining-smelting activities in Shuikoushan, Hunan Province, China. AB - The spatial variation of heavy metals in the soils in Shuikoushan mining-smelting area, Hunan Province, China, was investigated using multivariate and geo statistic analysis. A total of 106 composite soil samples were collected in an area of about 100 km2. Concentrations of total As, Cd, Pb, Zn, Cu and Cr were measured using inductively coupled plasma mass spectrometry (ICP-MS). Arsenic and Pb were found to have a common source, indicating the same sources and spreading processes, such as aerosols and airborne particulates from smelting chimneys. Airborne sources from smelting chimneys contributed greatly to Cd in the area, which demonstrated the same dispersion pattern as As and Pb. However, two hot spots of Cd around smelters were possibly enlarged by wastewaters, demonstrating another important source of Cd in Shuikouhsan. Geo-statistic interpolated mapping demonstrated that hot-spots of Zn were only found proximal to the large smelters, suggesting that Zn primarily came from the chimneys of larger smelters. The major Cu hot-spots appeared closely to the tailing dam, indicating that weathering and leaching of tailings were the major sources of Cu contamination in Shuikoushan. Our findings indicated that airborne volatile particles and aerosols contributed the most to As, Cd, Pb, Zn and Cu contamination, while Cd and Cu may also derive from the discharge of wastewater from smelters and the leaching of tailings, respectively. PMID- 19999972 TI - Performance of microbiological control by a point-of-use filter system for drinking water purification. AB - Purification capacity of a faucet mounted type water filter for home use was evaluated, particularly with regard to microbiological performance under different running conditions. Biofilms were formed inside the filter, affecting the bacterial quality of the effluent water. Low flow rate, long stagnation period and high filter temperature were found favorable for bacterial growth inside. By commercial analytical profile index (API) kits, ten different bacterial species were identified in drinking water, four of which were probably contributed to the biofilm formation since they were also present in the biofilm. Fluorescence in situ hybridization (FISH) was used to confirm the API identification results, and direct viable count (DVC) method was employed to improve the sensitivity of FISH for the isolated Acinetobacter spp. and Pseudomonas putida as models. Relationship between the filter operating condition and the bacterial community alteration was partly revealed, which could provide the basic knowledge for the filter design and its practical use. PMID- 19999973 TI - Isolation and characterization of ferrous- and sulfur-oxidizing bacteria from Tengchong solfataric region, China. AB - Microbial oxidation and reduction of iron and sulfur are important parts of biogeochemical cycles in acidic environments such as geothermal solfataric regions. Species of Acidithiobacillus and Leptospirillum are the common ferrous iron and sulfur oxidizers from such environments. This study focused on the Tengchong sofataric region, located in Yunnan Province, Southwest China. Based on cultivation, 9 strains that grow on ferrous-iron and sulfuric compounds were obtained. Analysis of 16S rRNA genes of the 9 strains indicated that they were affiliated to Acidithiobacillus, Alicyclobacillus, Sulfobacillus, Leptospirillum and Acidiphilium. Physiological and phylogenetic studies indicated that two strains (TC-34 and TC-71) might represent two novel members of Alicyclobacillus. Strain TC-34 and TC-71 showed 94.8%-97.1% 16S rRNA gene identities to other species of Alicyclobacillus. Different from the previously described Alicyclobacillus species, strains TC-34 and TC-71 were mesophilic and their cellular fatty acids do not contain omega-cyclic fatty acids. Strain TC-71 was obligately dependent on ferrous-iron for growth. It was concluded that the ferrous-iron oxidizers were diversified and Alicyclobacillus species were proposed to take part in biochemical geocycling of iron in the Tengchong solfataric region. PMID- 19999974 TI - Isolation and characterization of Sporobolomyces sp. LF1 capable of degrading chlorimuron-ethyl. AB - A yeast strain which was capable of degrading sulfonylurea herbicide chlorimuron ethyl named as LF1 was isolated from a chlorimuron-ethyl contaminated soil near the warehouse of the factory producing chlorimuron-ethyl in Shenyang City, Northeast China. The strain was identified as Sporobolomyces sp., based on its morphological and physiological characteristics and the phylogenetic analysis of 18S rRNA gene sequence. So far, this is the only yeast strain of Sporobolomyces sp. which is able to degrade chlorimuron-ethyl. Incubation tests showed that when the initial concentration of chlorimuron-ethyl in culture was 5 mg/L, LFI could degrade more than 77% of the herbicide after incubation for 4 d at 30 degrees C. The possible mechanism of chlorimuron-ethyl degradation by LF1 could be the acidic hydrolysis caused by the acids from the metabolism of the yeast strain. Further study should be conducted to examine the pathways of chlorimuron-ethyl degradation by LF1 and to approach the feasibility of using LF1 to degrade the chlorimuron-ethyl in soil system. PMID- 19999975 TI - Characteristics of the microbial communities in the integrated vertical-flow constructed wetlands. AB - Microorganisms play an important role in removing pollutants from constructed wetlands. We investigated the microbial characteristics in a novel integrated vertical-flow constructed wetland (IVCW), which has been in operation in Wuhan, China since 1998. We used phospholipid fatty acid (PLFA) and amoA gene to analyze the structure and diversity of the microbial community within the IVCW. PLFA results suggested that the amount of bacterial PLFA was significantly higher than that of fungal PLFA, but the total microbial biomass represented by PLFA index was low in the system. Microbial spatial distribution showed significantly higher bacterial (both G+ and G-) and fungal biomass in the surface than in the subsurface layers. The ratios of monounsaturated to branched PLFA demonstrated that an anaerobic layer sandwiched by two aerobic layers existed in the IVCW, consistent with the redox potential results. Analysis of the amoA revealed the presence of Nitrosomonas-like sequences in the surface substrate of the downflow chamber and apparent diversities of ammonia-oxidizing bacteria in the system. These results suggest that microorganisms, despite their relatively low biomass, have inhabited the IVCW, and the results will offer some valuable information on microbe to system designers and managers. PMID- 19999976 TI - Phytoplankton community structure and environmental parameters in aquaculture areas of Daya Bay, South China Sea. AB - Environmental characteristics and phytoplankton community structure were investigated in two aquaculture areas in Dapeng Cove of Daya Bay, South China Sea, between April 2005 and June 2006. Phytoplankton abundance ranged between 5.0 and 8877.5 cells/mL, with an average of 751.8 cells/mL. The seasonal cycle of phytoplankton were demonstrated by frequent oscillations, with recurrent high abundances from late spring to autumn and a peak stage in late winter. Diatoms were the predominant phytoplankton group, accounting for 93.21% of the total abundance. The next most abundant group was the dinoflagellates, which made up only 1.24% of total abundance. High concentrations of Alexandrium tamarense (Lebour) Balech with a maximum of 603.0 cells/mL were firstly recorded in this area known for high rates of paralytic shellfish poisoning (PSP) contamination. Temperatures and salinities were within the suitable values for the growth of phytoplankton, and were important in phytoplankton seasonal fluctuations. The operation of the Daya Bay Nuclear Power Station (DNPS) exerts influences on the phytoplankton community and resulted in the high abundances of toxic dinoflagellate species during the winter months. Dissolved inorganic nitrogen (DIN) and dissolved silicate (DSi) were sufficient, and rarely limited for the growth of phytoplankton. Dissolved inorganic phosphorus (DIP) was the most necessary element for phytoplankton growth. The enriched environments accelerated the growth of small diatoms, and made for the shift in predominant species from large diatom Rhizosolenia spp. to chain-forming diatoms such as Skeletonema costatum, Pseudo-nitzschia spp. and Thalassiosira subtilis. PMID- 19999977 TI - Optimizing aerobic biodegradation of dichloromethane using response surface methodology. AB - Response surface methodology (RSM) was employed to evaluate the optimum aerobic biodegradation of dichloromethane (DCM) in pure culture. The parameters investigated include the initial DCM concentration, glucose as an inducer and hydrogen peroxide as terminal electron acceptor (TEA). Maximum aerobic biodegradation efficiency was predicted to occur when the initial DCM concentration was 380 mg/L, glucose 13.72 mg/L, and H202 115 mg/L. Under these conditions the aerobic biodegradation rate reached up to 93.18%, which was significantly higher than that obtained under original conditions. Without addition of glucose, degradation efficiencies were < or = 80% at DCM concentrations < 326 mg/L. When concentrations of DCM were more than 480 mg/L, the addition of hydrogen peroxide did not help to significantly increase DCM degradation efficiency. When DCM concentrations increased from 240 to 480 mg/L, the overall DCM degradation efficiency decreased from 91% to 60% in the presence of H2O2 for 120 mg/L. PMID- 19999979 TI - Microwave thermal remediation of crude oil contaminated soil enhanced by carbon fiber. AB - Thermal remediation of the soil contaminated with crude oil using microwave heating enhanced by carbon fiber (CF) was explored. The contaminated soil was treated with 2.45 GHz microwave, and CF was added to improve the conversion of microwave energy into thermal energy to heat the soil. During microwave heating, the oil contaminant was removed from the soil matrix and recovered by a condensation system of ice-salt bath. The experimental results indicated that CF could efficiently enhance the microwave heating of soil even with relatively low dose. With 0.1 wt.% CF, the soil could be heated to approximately 700 degrees C within 4 min using 800 W of microwave irradiation. Correspondingly, the contaminated soil could be highly cleaned up in a short time. Investigation of oil recovery showed that, during the remediation process, oil contaminant in the soil could be efficiently recovered without causing significant secondary pollution. PMID- 19999978 TI - Oxidative stress and apoptosis of Carassius auratus lymphocytes induced by nonplanar (PCB153) and coplanar (PCB169) polychlorinated biphenyl congeners in vitro. AB - Among all the 209 kinds of polychlorinated biphenyls (PCBs) congeners, nonplanar and coplanar PCB congeners have different levels of toxicity on mammal cells such as neuronal cells, but little is known about their toxicity on fish cells although PCB congeners usually have high bioaccumulation abilities in the detected fish bodies. This study showed that 2,2',4,4',5,5'-hexacholorbiphenyl (PCB153, nonplanar congener) and 3,3',4,4',5,5'-hexacholorbiphenyl (PCB 169, coplanar congener) caused apoptosis on the isolated crucian carp (Carassius auratus) lymphocytes and the induced cytotoxicity was structure-dependent. According to the laser confocal microscope observations, apoptosis was clearly distinguished by condensation of nucleus, shrinkage and formation of apoptotic bodies. DNA fragmentation was detected by agarose gel electrophoresis. These typical morphological and biochemical characteristics indicate the occurrence of apoptosis on fish lymphocytes. According to the flow cytometry analysis, after the cells were exposed to 10 micromol/L PCBs for 3 h, the apoptotic percentage induced by PCB153 was 23.41%, while that induced by PCB169 was even higher (31.03%). Furthermore, incubating PCBs with fish lymphocytes enhanced levels of reactive oxygen species (ROS) and malondialdehyde (MDA), clearly indicating the presence of oxidative stress and lipid peroxidation. Our data also demonstrate that the different cytotoxic effects induced by coplanar and nonplanar PCBs were correlated with their structural characteristics and the coplanar congener was more cytotoxic than nonplanar congener. This study suggests that cytotoxicity mechanisms of the PCB congeners on fish lymphocytes depend on their planarity and chemical structures. PMID- 19999980 TI - Catalytic reduction of nitric oxide with carbon monoxide on copper-cobalt oxides supported on nano-titanium dioxide. AB - A series of copper-cobalt oxides supported on nano-titanium dioxide were prepared for the reduction of nitric oxide with carbon monoxide and characterized using techniques such as XRD, BET and TPR. Catalyst CuCoOx/TiO2 with Cu/Co molar ratio of 1/2, Cu-Co total loading of 30% at the calcination temperature of 350 degrees C formed CuCo2O4 spinel and had the highest activity. NO conversion reached 98.9% at 200 degrees C. Mechanism of the reduction was also investigated, N2O was mainly yielded below 100 degrees C, while N2 was produced instead at higher temperature. O2 was supposed to accelerate the reaction between NOx and CO for its oxidation of NO to give more easily reduced NO2, but the oxidation of CO by O2 to CO2 decreased the speed of the reaction greatly. Either SO2 or H2O had no adverse impact on the activity of NO reduction; however, in the presence of both SO2 and H2O, the catalyst deactivated quickly. PMID- 19999981 TI - Transformation of organic matters in fresh leachate during anaerobic degradation under long hydraulic retention time. AB - This study investigated the biodegradability of fresh leachate from a young municipal solid waste landfill at the inoculum to substrate ratios (V/V) of 10/100, 25/100, 50/100 and 100/100, as well as the transformation of organic matters in leachate under prolonged hydraulic retention time (HRT) of 50 d. Fresh leachate showed a good biodegradability, with chemical oxygen demand (COD) removal efficiency as high as 87%-92% and cumulative methane yield close to theoretical value (0.35 L CH4/g CODdeg). Methane production and COD depletion presented biphasic characteristics associated with the successive utilization of two major intermediates, acetate and propionate. The biphasic degradation of fresh leachate was resulted from the different hydrolysis rates of diverse substrates and the changes in microorganism community structure. After 50 d, the effluents were dominated by high-MW organic compounds (MW > 10 kDa) at each inoculum ratio, which might be refractory compounds released from cell lyses. PMID- 19999982 TI - Characterization of refuse landfill leachates of three different stages in landfill stabilization process. AB - Landfill leachates with different ages (mature leachate, 11 years; semi-mature leachate, 5 years; fresh leachate, under operation) were collected from Laogang Refuse Landfill, Shanghai to characterize the colloid size distribution and variations of leachate. These leachates were separated using micro-filtration and ultra-filtration into specific size fractions, i.e., suspended particles (SP) (> 1.2 microm), coarse colloids (CC) (1.2-0.45 microm), fine colloids (FC) (0.45 microm, 5 kDa/1 kDa molecular weight (MW)), and dissolved organic matters (DM, < 5 kDa/1 kDa MW). The specific colloids in each size fraction were quantified and characterized through chemical oxygen demands (COD), total solid (TS), pH, NH4+ N, total organic carbon (TOC) and fixed solid (FS). It was found that COD, NH4+-N and TS in leachate decreased significantly over ages, while pH increased. The dissolved fractions (< 5 kDa/1 kDa) dominated (over 50%) in three leachates in terms of COD, and the organic matter content in dissolved fraction of leachates decreased and the inorganic matter increased as the disposal time extended, with the TOC/COD ratio 30%-7%. Dissolved fractions decreased from 82% to 40% in terms of TOC as the disposal time extended, suggested that the organic matter remained in leachate would form into middle molecular weight substances during the degradation process. PMID- 19999983 TI - Influence of chlorine on methane oxidation. AB - Experiments on CH4/Cl2/O2/N2 oxidation were conducted in an atmospheric pressure flow reactor to understand the influence of chlorine on hydrocarbon oxidation in hazardous waste incineration. The reaction temperature varied from 973 to 1273 K and the chlorine to hydrogen mole ratio (Cl/H) of the inlet mixture varied from 0 to 0.44. The species produced in the reaction were measured online with Fourier transform infrared spectroscopy (FT-IR). It was found that the destruction and removal efficiency of CH4 increased with Cl/H mole ratio. Increasing Cl/H favored COCl2 and CO formation and inhibited the CO oxidation process. As Cl/H approached 0.44, the concentrations of CH2Cl2 and CH3Cl first increased, and then declined. Reaction temperature greatly affected the reaction system. Increasing temperatures raised the destruction removal efficiency of CH4 and decreased the concentrations of CH3Cl and CH2Cl2. With a certain ratio of Cl/H, the concentrations of CO and COCl2 first increased and then declined. The CO and COCl2 concentration peak was observed around 1100 K and 1023 K, respectively. When the reaction temperature exceeded 1273 K, carbon in CH4 was mostly converted to CO2. It could be concluded that the presence of chlorine enhanced the destruction of CH4, but resulted in the more toxic incomplete combustion products emission such as COCl2 when the reaction temperature was not high enough. PMID- 19999984 TI - Electrochemical incineration of dimethyl phthalate by anodic oxidation with boron doped diamond electrode. AB - The anodic oxidation of aqueous solutions containing dimethyl phthalate (DMP) up to 125 mg/L with sodium sulfate (Na2SO4) as supporting electrolyte within the pH range 2.0-10.0 was studied using a one-compartment batch reactor employing a boron-doped diamond (BDD) as anode. Electrolyses were carried out at constant current density (1.5-4.5 mA/cm2). Complete mineralization was always achieved owing to the great concentration of hydroxyl radical (*OH) generated at the BDD surface. The effects of pH, apparent current density and initial DMP concentration on the degradation rate of DMP, the specific charge required for its total mineralization and mineralization current efficiency were investigated systematically. The mineralization rate of DMP was found to be pH-independent and to increase with increasing applied current density. Results indicated that this electrochemical process was subjected, at least partially, to the mass transfer of organics onto the BDD surface. Kinetic analysis of the temporal change of DMP concentration during electrolysis determined by High Performance Liquid Chromatography (HPLC) revealed that DMP decay under all tested conditions followed a pseudo first-order reaction. Aromatic intermediates and generated carboxylic acids were identified by Gas Chromatography-Mass Spectrometry (GC-MS) and a general pathway for the electrochemical incineration of DMP on BDD was proposed. PMID- 19999985 TI - Effect of sludge retention time on sludge characteristics and membrane fouling of membrane bioreactor. AB - Three identical membrane bioreactors (MBRs) were operated over 2 years at different sludge retention time (SRT) of 10 d, 40 d and no sludge withdrawal (NS), to elucidate and quantify the effect of SRT on the sludge characteristics and membrane fouling. The hydraulic retention times of these MBRs were controlled at 12 h. With increasing SRT, the sludge concentrations in the MBRs increased, whereas the ratio of volatile suspended solid to the total solid decreased, and the size of sludge granule diminished in the meantime. A higher sludge concentration at long SRT could maintain a better organic removal efficiency, and a longer SRT was propitious to the growth of nitrifiers. The performance of these MBRs for the removal of COD and NH4+-N did not change much with different SRTs. However, the bioactivity decreased as SRT increase. The measurement of specific oxygen uptake rates (SOUR) and fluorescence in situ hybridization (FISH) with rRNA-targeted oligonucleotide probes testified that SOUR and the proportion of the bacteria-specific probe EUB338 in all DAPI-stainable bacteria decreased with increasing SRT. The concentrations of total organic carbon, protein, polysaccharides and soluble extracellular polymeric substance (EPS) in the mixed liquor supernatant also decreased with increasing SRT. The membrane fouling rate was higher at shorter SRT, and the highest fouling rate appeared at a SRT of 10 d. Both the sludge cake layer and gel layer had contribution to the fouling resistance, but the relative contribution of the gel layer decreased as SRT increase. PMID- 19999986 TI - Heterotrophic ammonium removal characteristics of an aerobic heterotrophic nitrifying-denitrifying bacterium, Providencia rettgeri YL. AB - Bacterium Providencia rettgeri YL was found to exhibit an unusual ability to heterotrophically nitrify and aerobically denitrify various concentrations of ammonium (NH4+-N). In order to further understand its removal ability, several experiments were conducted to identify the growth and ammonium removal response at different carbon to nitrogen (C/N) mass ratios, shaking speeds, temperatures, ammonium concentrations and to qualitatively verify the production of nitrogen gas using gas chromatography techniques. Results showed that under optimum conditions (C/N 10, 30 degrees C, 120 r/min), YL can significantly remove low and high concentrations of ammonium within 12 to 48 h of growth, respectively. The nitrification products hydroxylamine (NH2OH), nitrite (NO2(-)) and nitrate (NO3( )) as well as the denitrification product, nitrogen gas (N2), were detected under completely aerobic conditions. PMID- 19999987 TI - Preparation of high concentration polyaluminum chloride with high content of Alb or Alc. AB - A novel membrane distillation concentration method was used to prepare high concentration polyaluminum chloride (PACl) with high content of Alb or Alc. 2.52 mol/L PACl1 with 88% Alb and 2.38 mol/L PACl2 with 61% Alc were successfully prepared. Three coagulants, AlCl3, PACl1 and PACl2 were investigated on their hydrolysis behavior and speciation under different conditions. The effects of pH and dilution ratio on Al species distribution were investigated by ferron assay. Experimental result showed that pH had a significant effect on Al species distribution for the three coagulants. Dilution ratio had little effects on Alb and Alc distribution in whole dilution process except the beginning for PACl1 and PACl2. The results indicated that transformation of Al depends largely on their original composition. AlCl3 was the most unstable coagulant among these three coagulants during hydrolysis process. PACl1 and PACl2 with significant amounts of highly charged and stable polynuclear aluminum hydrolysis products were less affected by the hydrolysis conditions and could maintain high speciation stability under various conditions. PMID- 19999988 TI - Application of ferrate(VI) in the treatment of industrial wastes containing metal complexed cyanides : a green treatment. AB - Ferrate(VI) was employed for the oxidation of cyanide (CN) and simultaneous removal of copper or nickel in the mixed/complexed systems of CN-Cu, CN-Ni, or CN Cu-Ni. The degradation of CN (1.00 mmol/L) and removal of Cu (0.095 mmol/L) were investigated as a function of Fe(VI) doses from 0.3-2.00 mmol/L at pH 10.0. It was found that Fe(VI) could readily oxidize CN and the reduction of Fe(VI) into Fe(III) might serve efficiently for the removal of free copper ions. The increase in Fe(VI) dose apparently favoured the CN oxidation as well as Cu removal. Moreover, the pH dependence study (pH 10.0-13.0) revealed that the oxidation of CN was almost unaffected in the studied pH range (10.0-13.0), however, the maximum removal efficiency of Cu was obtained at pH 13.0. Similarly, treatment was carried out for CN-Ni system having the initial Ni concentration of 0.170 mmol/L and CN concentration of 1.00 mmol with Fe(VI) dose 2.00 mmol at various pH values (10.0-12.0). Results showed a partial oxidation of CN and partial removal of Ni. It can be observed that Fe(VI) can partially degrade the CN-Ni complex in this pH range. Further, Fe(VI) was applied for the treatment of simulated industrial waste/effluent waters treatment containing CN, Cu, and Ni. PMID- 19999989 TI - Secondary organic carbon quantification and source apportionment of PM10 in Kaifeng, China. AB - During 2005, the filter samples of ambient PM10 from five sites and the source samples of particulate matter were collected in Kaifeng, Henan Province of China. Nineteen elements, water-soluble ions, total carbon (TC) and organic carbon (OC) contained in samples were analyzed. Seven contributive source types were identified and their contributions to ambient PM10 were estimated by chemical mass balance (CMB) receptor model. Weak associations between the concentrations of organic carbon and element carbon (EC) were observed during the sampling periods, indicating that there was secondary organic aerosol pollution in the urban atmosphere. An indirect method of "OC/EC minimum ratio" was applied to estimate the concentration of secondary organic carbon (SOC). The results showed that SOC contributed 26.2%, 32.4% and 18.0% of TC in spring, summer-fall and winter, respectively, and the annual average SOC concentration was 7.07 microg/m3, accounting for 5.73% of the total mass in ambient PM10. The carbon species concentrations in ambient PM10 were recalculated by subtracting SOC concentrations from measured concentrations of TC and OC to increase the compatibility of source and receptor measurements for CMB model. PMID- 19999990 TI - Element and mineral characterization of dust emission from the saline land at Songnen Plain, Northeastern China. AB - Recent observations of Asian dust storms show an eastern expansion of the source area to degraded lands, where dust emissions have been little studied. The dust concentrations over the saline land of the western Songnen Plain (SSL), Northeastern China, are circumstantially higher than those from the northwestern Chinese deserts. These concentrations are sensitive to the surface soil conditions and wind velocity on the ground. The dust samples collected during dust storm events on the SSL contain abundant Na, Mg, Al, K, Ca, Fe and Ti, as well as toxic elements such as Cu, V, Zn and Ba. Individual particle analysis reveals that fine saline particles (< 10 microm in diameter) on the saline land, consisting largely of carbonate, halite and sulfate together with lithogenic minerals such as SiO2 and aluminosilicate, are eventually uplifted during the interval from spring to autumn. The predominantly fine saline particles uplifted from the SSL are likely transported eastward by the winter monsoon circulation and westerlies. Recent degradation of saline lands in Northeastern China would not only increase the frequency of dust storm events in the downwind area, but also might change the chemical composition of the Asian dust emissions. PMID- 19999991 TI - Characteristic of organic aerosol in a remote area of Okinawa Island. AB - A continuous investigation of aerosol pollutants in Cape Hedo, Japan was conducted from October 2005 to August 2006 by Aerodyne aerosol mass spectrometer (AMS). This article focused on the analysis of long-range transport of organic aerosol from continental origin to the remote island. Based on the transport distance and air mass origin, four main air mass origins were considered, including North China, South China, Japan and Korea. Although the mass concentration and air mass origin were quite different during study period, the mass spectrum and size distribution of organic matter and oxidized organics were similar, which showed uni-modal distribution with modal diameter of around 500 nm. The loss rate of organics was (5.15 +/- 0.55) x 10(-6) s(-1) obtained by plotting the concentration against the transport time. Conversion rate of SO2 to sulfate and oxidation rate of organics were (1.07 +/- 0.15) x 10(-5) and (1.09 +/ 0.52) x 10(-6) s(-1), respectively. PMID- 19999992 TI - Effect of biosurfactant on the sorption of phenanthrene onto original and H2O2 treated soils. AB - The objective of this study was to examine the effect of biosurfactant on the sorption of phenanthrene (PHE) onto the original or H2O2-treated black loamy soil (typic isohumisols) and red sandy soil (typic ferralisols). The sorption isotherms were performed with the original and "soft" carbon-removed soils in the presence and absence of biosurfactant (200 mg/L). The sorption and degradation of biosurfactant were investigated. The result showed that organic matter played an important role in PHE sorption onto the black loamy and red sandy soils, and the PHE sorption isotherms on the "soft" carbon-removed soils exhibited more nonlinearity than those on the original soils. The values of partition coefficient (Kd) on the original black loamy soil with or without 200 mg/L biosurfactant were 181.6 and 494.5 mL/g, respectively. Correspondingly, in the red sandy soil, Kd was 246.4 and 212.8 mL/g in the presence or absence of biosurfactant, respectively. The changes of Kd suggested that biosurfactant inhibited PHE sorption onto the black loamy soil, but facilitated PHE sorption onto the red sandy soil. The nonlinearity of PHE sorption isotherm was decreased in the presence of biosurfactant. Site specific sorption might occur during PHE sorption onto both the original and the "soft" carbon-removed soils in the presence of biosurfactant. It was noted that biosurfactant could also be sorbed onto soils. The maximal sorption capacity of the red sandy soil for biosurfactant was (76.9 +/- 0.007) microg/g, which was 1.31 times that of black loamy soil. Biosurfactant was degraded quickly in the two selected soils, and 92% of biosurfactant were mineralized throughout the incubation experiment for 7 d. It implied that biosurfactant should be added frequently when the remediation of polycyclic aromatic hydrocarbon (PAH)-contaminated soils was conducted through PAH desorption approach facilitated by biosurfactant. PMID- 19999993 TI - Levels, distributions, and source identification of organochlorine pesticides in the topsoils in Northeastern China. AB - Seventeen topsoil samples (9 urban, 4 suburban, 3 rural and 1 background) were collected in/around Harbin, a typical city in northeast of China, to measure concentration levels of organocholrine pesticides (OCPs) in topsoil of Northeastern China in 2006. Hexachlorohexanes (HCH), dichlorodiphenyltrichloroethane (DDT), and hexachlorobenzene (HCB) were detected in soil samples with mean concentrations (in pg/g dry weight (dw)) of 7120, 5425, and 1039, respectively. The mean concentrations for other OCPs were very low, 4.8 pg/g dw for chlordane and 3.3 pg/g dw for endosulfan. Source identification analysis reveals that all OCPs found in soil samples were due to historical use of these chemicals or from other source regions through long- and short-range atmospheric transport. DDT was mainly used in the rural sites, whereas the sources of HCB, chlordane and endosulfan were mainly in the urban area. HCH was found almost equally in both urban and rural area. Soil concentrations of all detected OCPs, except HCHs, in and around Harbin were much lower than those in the southeast of China, which is expected since the use of these OCPs in the former was much lower than that in the latter, however higher HCH concentrations in and around Harbin than those found in most places of the Southeast China is not expected. It is suggested that high HCH concentration in soil of Northeast China was most likely due to long-range atmospheric transport (LRAT) from Southeast China and the cold condensation process. PMID- 19999994 TI - Effects of elevated CO2 concentration and nitrogen supply on biomass and active carbon of freshwater marsh after two growing seasons in Sanjiang Plain, Northeast China. AB - An experiments were carried out with treatments differing in nitrogen supply (0, 5 and 15 g N/m2) and CO2 levels (350 and 700 micromol/mol) using OTC (open top chamber) equipment to investigate the biomass of Calamagrostis angustifolia and soil active carbon contents after two years. The results showed that elevated CO2 concentration increased the biomass of C. angustifolia and the magnitude of response varied with each growth period. Elevated CO2 concentration has increased aboveground biomass by 16.7% and 17.6% during the jointing and heading periods and only 3.5% and 9.4% during dough and maturity periods. The increases in belowground biomass due to CO2 elevation was 26.5%, 34.0% and 28.7% during the heading, dough and maturity periods, respectively. The responses of biomass to enhanced CO2 concentrations are differed in N levels. Both the increase of aboveground biomass and belowground biomass were greater under high level of N supply (15 g N/m2). Elevated CO2 concentration also increased the allocation of biomass and carbon in root. Under elevated CO2 concentration, the average values of active carbon tended to increase. The increases of soil active soil contents followed the sequence of microbial biomass carbon (10.6%) > dissolved organic carbon (7.5%) > labile oxidable carbon (6.6%) > carbohydrate carbon (4.1%). Stepwise regressions indicated there were significant correlations between the soil active carbon contents and plant biomass. Particularly, microbial biomass carbon, labile oxidable carbon and carbohydrate carbon were found to be correlated with belowground biomass, while dissolved organic carbon has correlation with aboveground biomass. Therefore, increased biomass was regarded as the main driving force for the increase in soil active organic carbon under elevated CO2 concentration. PMID- 19999995 TI - Distribution characteristics of total mercury and methylmercury in the topsoil and dust of Xiamen, China. AB - The levels and distribution of mercury (Hg) species, including total mercury (THg) and methylmercury (MeHg) in the topsoil and dust collected from twenty sampling stations located in different land function areas of Xiamen, China, were investigated. The THg concentrations in topsoil ranged from 0.071 to 1.2 mg/kg, and in dust ranged from of 0.034 to 1.4 mg/kg. For stations where the THg of dust was less than 0.31 mg/kg, THg concentrations in the topsoil were significantly correlated to those in the corresponding dust (r = 0.597, n = 16, P = 0.014). The MeHg concentrations in topsoil were varied between 0.14 and 5.7 microg/kg. The ratios of MeHg/THg in the topsoil ranged from 0.069% to 0.74%. The range of MeHg concentration in the dust were 0.092-2.3 microg/kg. The ratios of MeHg/THg in the dust were at the same level as those in the topsoil. The MeHg concentrations in both topsoil and dust were linked to corresponding THg concentrations and soil organic matter. Neither THg nor MeHg concentration in the topsoil and dust was obviously linked to the land function. PMID- 19999996 TI - Bioaccumulation of heavy metals by Phragmites australis cultivated in synthesized substrates. AB - Accumulation of heavy metals from various oxides with adsorbed cadmium by wetland plant Phragmites australis was studied to evaluate the fate of heavy metals in the sediment of constructed wetlands. Hoagland solution was used as nutrition supply, and single metal oxide with adsorbed cadmium was applied as contaminant to study the accumulation characteristics of cadmium and the substrate metals by P. australis. After 45-d treatment, the bioaccumulation degree in root followed the order: Al(OH)3 > Al2O3 > Fe3O4 > MnO2 > FeOOH. Heavy metals absorbed by P. australis were largely immobilized by the roots with little translocation to aboveground parts. PMID- 19999997 TI - Phosphorus utilization and microbial community in response to lead/iron addition to a waterlogged soil. AB - Constructed wetlands have emerged as a viable option for helping to solve a wide range of water quality problems. However, heavy metals adsorbed by substrates would decrease the growth of plants, impair the functions of wetlands and eventually result in a failure of contaminant removal. Typha latifolia L., tolerant to heavy metals, has been widely used for phytoremediation of Pb/Zn mine tailings under waterlogged conditions. This study examined effects of iron as ferrous sulfate (100 and 500 mg/kg) and lead as lead nitrate (0, 100, 500 and 1000 mg/kg) on phosphorus utilization and microbial community structure in a constructed wetland. Wetland plants (T. latifolia) were grown for 8 weeks in rhizobags filled with a paddy soil under waterlogged conditions. The results showed that both the amount of iron plaque on the roots and phosphorus adsorbed on the plaque decreased with the amount of lead addition. When the ratio of added iron to lead was 1:1, phosphorus utilized by plants was the maximum. Total amount of phospholipids fatty acids (PLFAs) was 23%-59% higher in the rhizosphere soil than in bulk soil. The relative abundance of Gram-negative bacteria, aerobic bacteria, and methane oxidizing bacteria was also higher in the rhizosphere soil than in bulk soil, but opposite was observed for other bacteria and fungi. Based on cluster analysis, microbial communities were mostly controlled by the addition of ferrous sulfate and lead nitrate in rhizosphere and bulk soil, respectively. PMID- 19999998 TI - Infiltration characteristics of non-aqueous phase liquids in undisturbed loessal soil cores. AB - The widespread contamination of soils and aquifers by non-aqueous phase liquids (NAPL), such as crude oil, poses serious environmental and health hazards globally. Understanding the infiltration characteristics of NAPL in soil is crucial in mitigating or remediating soil contamination. The infiltration characteristics of crude and diesel oils into undisturbed loessal soil cores, collected in polymethyl methacrylate cylindrical columns, were investigated under a constant fluid head (3 cm) of either crude oil or diesel oil. The infiltration rate of both crude and diesel oils decreased exponentially as wetting depth increased with time. Soil core size and bulk density both had significant effects on NAPL infiltration through the undisturbed soil cores; a smaller core size or a greater bulk density could reduce oil penetration to depth. Compacting soil in areas susceptible to oil spills may be an effective stratage to reduce contamination. The infiltration of NAPL into soil cores was spatially anisotropic and heterogeneous, thus recording the data at four points on the soil core is a good stratage to improve the accuracy of experimental results. Our results revealed that crude and diesel oils, rather than their components, have a practical value for remediation of contaminated loessal soils. PMID- 19999999 TI - Removal of metals by sorghum plants from contaminated land. AB - The growth of high biomass crops facilitated by optimal of agronomic practices has been considered as an alternative to phytoremediation of soils contaminated by heavy metals. A field trial was carried out to evaluate the phytoextraction efficiency of heavy metals by three varieties of sweet sorghum (Sorghum biocolor L.), a high biomass energy plant. Ethylene diamine tetraacetate (EDTA), ammonium nitrate (NH4NO3) and ammonium sulphate ((NH4)2SO4) were tested for their abilities to enhance the removal of heavy metals Pb, Cd, Zn, and Cu by sweet sorghum from a contaminated agricultural soil. Sorghum plants always achieved the greatest removal of Pb by leaves and the greatest removal of Cd, Zn and Cu by stems. There was no significant difference among the Keller, Rio and Mray varieties of sweet sorghums in accumulating heavy metals. EDTA treatment was more efficient than ammonium nitrate and ammonium sulphate in promoting Pb accumulation in sweet sorghum from the contaminated agricultural soil. The application of ammonium nitrate and ammonium sulphate increased the accumulation of both Zn and Cd in roots of sorghum plants. Results from this study suggest that cropping of sorghum plants facilitated by agronomic practices may be a sustainable technique for partial decontamination of heavy metal contaminated soils. PMID- 20000000 TI - Long-term impact of municipal sewage irrigation on treated soil and black locust trees in a semi-arid suburban area of Iran. AB - The effects of municipal sewage irrigation on the soil and black locust (Robinia pseudoacacia L.) tree were studied. For this purpose, two artificial black locust stands under irrigation of municipal sewage and well water were selected in south of Tehran, Iran. Data were collected using technique of systematic random sampling with 4 replicates in each stand. It was found that the growth of black locust tree, as indicated by diameter at breast height, total height, crown length, average crown diameter, basal area and volume, in sewage irrigation stand was much higher than that of well water irrigation stand (P < 0.01). Plant analysis indicated that concentrations of leaf nutrients (N, P, K, Ca, Mg, Na, Fe, Mn, Cu and Zn) were greater in sewage-irrigated trees, without toxicity to the minerals of tree leaf, than those of well waterirrigated trees, and positively correlated with their respective value in soil. Ni, Cr and Pb were not detected in leaf samples. Application of sewage resulted in a 1.5-fold increase in the concentrations of soil nutrients, Ni, Cr and Pb. Among these minerals only Pb and Ni in some soil samples exceeded the toxicity limit. The increase in pH, electrical conductivity (EC) and organic carbon of soil was also observed in sewage irrigation. Results confirm that besides the use as irrigation water, municipal sewages are also a potential source of plant nutrients. However, significant accumulation of heavy metals such as Pb and Ni in soil needs to be monitored. PMID- 20000001 TI - Isolation of marine benzo[a]pyrene-degrading Ochrobactrum sp. BAP5 and proteins characterization. AB - A bacterial strain BAP5 with a relatively high degradation ability of benzo[a]pyrene (BaP) was isolated from marine sediments of Xiamen Western Sea, China and identified as Ochrobactrum sp. according to 16S rRNA gene sequence as well as Biolog microbial identification system. Strain BAP5 could grow in mineral salt medium with 50 mg/L of BaP and degrade about 20% BaP after 30 d of incubation. Ochrobactrum sp. BAP5 was able to utilize other polycyclic aromatic hydrocarbons (PAHs) (such as phenanthrene, pyrene and fluoranthene) as the sole carbon source and energy source, suggesting its potential application in PAHs bioremediation. The profile of total soluble protein from Ochrobactrum sp. BAP5 was also investigated. Some over- and special-expressed proteins of strain BAP5 when incubated with the presence of BaP were detected by two-dimensional polyacrylamide gel electrophoresis, and found to be related with PAHs metabolism, DNA translation, and energy production based on peptide fingerprint analysis through matrix-assisted laser desorption/ionization-time of flight mass spectrometry. PMID- 20000002 TI - Microbiological parameters and maturity degree during composting of Posidonia oceanica residues mixed with vegetable wastes in semi-arid pedo-climatic condition. AB - The aim of this study was to characterize the biological stability and maturity degree of compost during a controlled pile-composting trial of mixed vegetable residues (VR) collected from markets of Tunis City with residues of Posidonia oceanica (PoR), collected from Tunis beaches. The accumulation in beaches (as well as their removal) constitutes a serious environmental problem in all Mediterranean countries particularly in Tunisia. Aerobic-thermophilic composting is the most reasonable way to profit highly-valuable content of organic matter in these wastes for agricultural purposes. The physical, chemical, and biological parameters were monitored during composting over 150 d. The most appropriate parameters were selected to establish the maturity degree. The main result of this research was the deduction of the following maturity criterion: (a) C/N ratio < 15; (b) NH4+-N < 400 mg/kg; (c) CO2-C < 2000 mg CO2-C/kg; (d) dehydrogenase activity < 1 mg TPF/g dry matter; (e) germination index (GI) > 80%. These five parameters, considered jointly are indicative of a high maturity degree and thus of a high-quality organic amendment which employed in a rational way, may improve soil fertility and soil quality. The mature compost was relatively rich in N (13.0 g/kg), P (4.74 g/kg) and MgO (15.80 g/kg). Thus composting definitively constitutes the most optimal option to exploit these wastes. PMID- 20000003 TI - Effect of sub-acute exposure to TiO2 nanoparticles on oxidative stress and histopathological changes in Juvenile Carp (Cyprinus carpio). AB - Increasing application of nanotechnology highlights the need to clarify and understand nanotoxicity. Mammalian and in vitro studies have raised concerns about the toxicity of titanium dioxide nanoparticles (TiO2-NPs), but there are limited data on ecotoxicity to aquatic organisms. In this work, the sub-acute toxicity of TiO2-NPs to carp (Cyprinus carpio) was assessed. Superoxide dismutase (SOD), catalase (CAT) and peroxidase (POD) activities and lipid peroxidation (LPO) levels in liver, gill and brain tissues of carps varied with concentration of TiO2-NPs suspensions and exposure time (up to 8 d). As a result, 100 and 200 mg/L TiO2-NPs caused statistically significant decrease in SOD, CAT and POD activities and significant increase in LPO levels in tissues (P < 0.05), suggesting that the fish exposed to these two concentrations of TiO2-NPs suffered from the oxidative stress. The extent of depletion of antioxidant enzymes activities and the elevation of LPO in the liver was the greatest, indicating that the liver might be the most susceptible organ to TiO2-NPs exposure. In addition, carps had gill pathologies including edema and thickening of gill lamellae as well as gill filaments, and liver pathologies including necrotic and apoptosis hepatocytes after exposed to 100 and 200 mg/L TiO2-NPs for 20 d. These results indicated a potential risk from TiO2-NPs released into the aqueous environment. PMID- 20000004 TI - Acute effects of chlorpyryphos-ethyl and secondary treated effluents on acetylcholinesterase and butyrylcholinesterase activities in Carcinus maenas. AB - The acute effects of commercial formulation of chlorpyrifos-ethyl (Dursban) and the secondary treated industrial/urban effluent (STIUE) exposure on acetylcholinesterase (AChE) and butyrylcholinesterase (BuChE) activities in hepatopancreas and gills of Mediterranean crab Carcinus maenas were investigated. After 2 d of exposure to chlorpyriphos-ethyl, the AChE activity was inhibited in both organs at concentrations of 3.12 and 7.82 microg/L, whereas the BuChE was inhibited only at higher concentration 7.82 microg/L of commercial preparation Dursban. The exposure of crabs to Dursban (3.12 microg/L) showed a significant decrement of AChE activity at 24 and 48 h, whereas the BuChE was inhibited only after 24 h and no inhibition for both enzymes was observed after 72 h. Moreover, a significant repression of AChE activity was observed in both organs of C. maenas exposed to 5% of STIUE. Our experiments indicated that the measurement of AChE activity in gills and hepatopancreas of C. maenas would be useful biomarker of organophosphorous (OP) and of neurotoxic effects of STIUE in Tunisia. PMID- 20000005 TI - Potent antioxidant and radical-scavenging activity of Chenpi--compensatory and cooperative actions of ascorbic acid and citric acid. AB - Dried peels of Satsuma mandarin (Citrus unshiu Marcov.) have been used as traditional Chinese and Japanese medicine, which are called 'Chenpi'. In our present study, cold and hot water extracts of Chenpi exhibited a strong inhibitory activity against linoleic acid peroxidation and 1,1-diphenyl-2-picryl hydrazyl (DPPH) radical-scavenging activity compared with o-methanol extract. When these extracts were separated into ethanol-soluble(ES) and ethanol precipitable fractions, the dominant antioxidant and radical-scavenging activities were detected in the ES fractions, which suggests that these antioxidant and radical-scavenging activities are responsible for water-soluble and low-molecular-weight substances. As possible active principles for antioxidant activities in the water extracts, the contents of ascorbic acid and citric acid in these extracts were measured, and the antioxidant and radical scavenging activities of these substances were assayed at various concentrations. The experimental results indicate that the antioxidant activity against lipid peroxidation in the water extracts is dominantly associated with citric acid, and the DPPH radical-scavenging activity of the water extracts is majorly responsible for ascorbic acid, suggesting a compensatory action of ascorbic acid and citric acid in expression of the antioxidant and radical-scavenging activities of Chenpi. PMID- 20000006 TI - Vascular responses in glabrous and nonglabrous skin during acute mental stress in physically trained humans. AB - Acute mental stress induces sympathetic activation and influences vasomotor control in various organs. In the present study, to better understand the effect of physical training on peripheral vasomotor control during acute mental stress, we compared the skin vascular responses to mental arithmetic (MA) in physically trained and untrained humans. Eight physically trained (T group) and eight untrained (UT group) healthy volunteers performed 2 min of MA aloud in the supine position under a thermoneutral condition (28 degrees C). Skin blood flow (laser Doppler flowmetry) and local temperature were monitored at the glabrous (palm, sole) and nonglabrous (forearm, calf) sites. Cutaneous vascular conductance (CVC) was evaluated from the ratio of blood flow to mean arterial pressure (tonometry). Local sweating rate (SR) was measured in the sole and calf by the ventilated capsule method. In the T group, the CVC at glabrous sites consistently decreased (P < 0.05) during MA, while in the UT group, the stress-induced decreases in CVC were transient and gradually recovered during MA. The patterns of changes in CVC at the nonglabrous sites were substantially similar to those at the glabrous sites, but the decreases in CVC at the nonglabrous sites were smaller (P < 0.05) than those at the glabrous sites in both groups. Local temperature at the glabrous sites (especially in the sole) showed higher (P < 0.05) values in the T group compared with the UT group. The SR in the sole and calf were increased (P < 0.05) during MA but did not differ between the two groups. These findings suggest that physical training acts to heighten skin temperature at the glabrous sites but not at the nonglabrous sites. It is also suggested that the change of skin temperature by physical training modifies sympathetic vasomotor control in glabrous and nonglabrous skin during acute mental stress in the peripheral level. PMID- 20000007 TI - Pulmonary capillary hemangiomatosis in chronic cardiac failure due to aortic stenosis. AB - We report a case of pulmonary capillary hemangiomatosis in a patient with aortic stenosis. An 86-year-old Japanese female with chronic heart failure due to aortic stenosis suddenly died during hemodialysis. At an autopsy, severe aortic stenosis and cardiomegaly with both left and right ventricular hypertrophy were noted. In the lung, a diffuse proliferation of capillaries in the thickened alveolar septum and collections of hemosiderin-laden macrophages in the alveolar space were observed. These indicated that long-standing passive congestion from aortic stenosis might result in the occurrence of pulmonary capillary hemangiomatosis. PMID- 20000008 TI - [Possibility of a description of mind in terms of quantum theory]. AB - The problem of mind and consciousness cannot be treated as an object of science without much difficulty. Inevitably there is the self-contradiction that the work itself in science and thinking is an object of thought. In this meaning, we need to get back to the fundamental problem of how the world is perceived. The fundamental concept of quantum theory includes essentially unsolved problems such as the observation problem. In this paper, we begin to consider this fundamental problem in quantum theory and discuss the possibility of a description of mind in terms of quantum theory. PMID- 20000009 TI - [Autopsy case of drowning caused by accidental carbon dioxide intoxication in a hold tank]. AB - A 49-year-old male captain fell and unfortunately died in a hold tank where he had entered to rescue his fainting co-worker on the disposing waste fluid left there. An autopsy revealed that the captain died from drowning in the waste fluid. In order to clarify the cause of their falling in the tank, the gas in the hold tank was analyzed. The concentration of oxygen was 18.86 to 19.31%, carbon dioxide was 7.28 to 9.07% and the other gases, including hydrogen sulfide, were assessed to be under the normal level. It was concluded that the intoxication of carbon dioxide generated from the waste fluid fermentation was the cause of this fatal accident through loss of consciousness. It is necessary to recognize that carbon dioxide is a dangerous and deleterious gas in circumstances where the gas can be produced. PMID- 20000010 TI - [Palliative rehabilitation of two patients with terminal stage cancer: a visit to patient's home and the provision of advice by a nurse and rehabilitation staff members before discharge--from physical therapist's perspective]. AB - We report two patients with terminal stage cancer who spent some days at their home after a physical therapist, occupational therapist, nurse, and medical social worker all visited the patients' homes and advised the patients' family in regard to the appropriate care before the patients were discharged as a strategy for palliative rehabilitation. Case 1: A patient suffering from terminal stage cancer was bed-ridden because of a pathological fracture of the femur. After palliative rehabilitation, the patient was able to get out of the bed and improved her daily living activity level through physical therapy. She spent some days at home according to the results of the pre-discharge home visit guidance to her family. Case 2: A patient suffering from terminal stage cancer manifested symptoms of fatigue and generalized muscular weakness. After palliative rehabilitation, her muscle strength and physical endurance were improved by physical therapy and adjustment of the bed height. Because she was eager to go home, we took her to her home before being discharged, and she was able to spend a few hours at home. Pre-discharge home visit guidance by a nurse and rehabilitation staff members to the patient' s family in regard to appropriate home care may therefore be a good means of satisfying such patients' desire to see their home once more and thereby improve their quality of life. PMID- 20000011 TI - [Effectiveness of learning of nursing-work as early exposure by the 1st year medical students at a university hospital]. AB - We conducted a half day program included in the subject of "nursing and care" as early exposure at clinical sites for the 1st year medical students at a university hospital. This program aimed at understanding what nursing is through visit for study and what patients expect through doctor-patient communication. In order to evaluate the program and to clarify problems to be solved, comments and impressions reported by the medical students were analyzed qualitatively and inductively. As a result, we found that the students recognized the importance of communication with patients and of mental care for them. As for nursing, the students also realized the characteristics and significance of nursing. It is noteworthy that they acquired clear images of a medical doctor, including their roles in team-based medical care. We conclude that this program of early exposure to clinical sites is instructive for 1st year medical students. PMID- 20000012 TI - [Relationship between nurses' learning motivation inside/outside the workplace and job/ life satisfaction]. AB - For the purpose of clarifying the relationship between job, life-satisfaction and motivation for learning of nurses, a questionnaire survey by mailing method was conducted for 123 nurses (52.6% response rate) within the Kyushu area. The analysis was performed by comparing the mean value of scores for each satisfaction category and that of comprehensive satisfaction (hereafter refered to as satisfaction level) which were extracted by principal component analysis. It has been proved that the higher was the motivation for learning, the greater was the job satisfaction level (0.16 +/- 0.95 in H group; -0.56 +/- 0.99 in L group, P = 0.001) in the case of learning in the workplace, and that motivation for learning was decreased by their sentiment that their job and learning weren't evaluated properly or by their discontent they felt in participating in learning on a day-off and during off-hours. It has also been proved in the case of off-the job learning that the higher was the motivation for learning, the greater was the job satisfaction level (0.10 +/- 1.01 in H group; -0.35 +/- 0.90 in L group, P = 0.040), and that their motivation for learning was decreased by feeling that their job and learning weren't evaluated properly or that they weren't satisfied with their physical and mental health. Also in other learnings, the higher was the motivation for learning, the greater was the job satisfaction level (0.08 +/ 0.96 in H group; -0.27 +/- 1.11 in L group, P = 0.034), and the motivation for learning was decreased depending on the type of work, work-hours, and on the strain felt to be time-consuming. PMID- 20000013 TI - Maternal morbidity and mortality in peri-urban Kenya--assessing progress in improving maternal healthcare. AB - OBJECTIVE: Improving quality of maternal healthcare delivery is pivotal to reversing the trend of maternal health in developing countries and consequently, achieving the 5th millennium goal of reducing maternal mortality and improving universal access to reproductive health. Periodic facility-based reviews of maternal morbidity and mortality are crucial to improving quality of care delivered. METHODS: This retrospective study was conducted in PCEA Kikuyu Hospital, Kenya. Data on sociodemographics, recorded antenatal care activities, maternal morbidities and deaths were elicited from case notes of all pregnancies and births over a 2 year period and subsequently analyzed. RESULTS: There were 1716 deliveries during the period. 1405 (91%) of the women booked for antenatal care; 1068 (57.3%) had at least 3 visits before delivery. 842 (45.9%) were either untested for Human Immunodeficiency Virus or their results were not documented; 55 (3%) tested positive. The caesarean section rate was 27.4% (n=470); PMTCT (Prevention of Mother to Child Transmission) constituted 1.6% (10) of the indications. Urinary tract infection was the commonest maternal illness in pregnancy (14.5%, n=270), however, it was not found to be associated with any adverse outcome. Genital tract trauma was the commonest morbidity suffered by the women during delivery (90.6%, n=800). Psychological disorders constituted 5.3% (7) of reported postpartum complications. There were 2 maternal deaths out of 1673 live births, giving a maternal mortality ratio of 119.55 per 100,000 live births. The deaths resulted from septic abortion and eclampsia. CONCLUSION: Urinary tract infection is an important cause of maternal illness during pregnancy. Routine screening is recommended. PMTCT needs further consolidation to increase the number of screened women and ensure those who need interventions like caesarean sections get access to them. Diagnosis and management of psychological disorders in pregnancy remain unsatisfactory. Further studies to identify the true burden of these conditions are needed. PMID- 20000014 TI - Knowledge, attitude and practice toward HIIV/AIDS among the Moran population living in Dar-es-Salaam. AB - The socio-cultural system holding the Wamasai as a unique ethnic group in Tanzania is gradually declining, resulting in the young Masai warriors (Moran) to move away and seek jobs in urban centres without permission of the elders. Living and working in urban centres increases their chances of exposure to risk of contracting HIV/AIDS, which upon their return home will lead to the infection of other Wamasai in their social interaction especially during initiation ceremonies. The aim of the study was to assess knowledge, attitudes and practices towards HIV/AIDS prevention among the Moran who have migrated to Dar es Salaam City. A cross sectional study was carried out from July to September 2006 in Dar es-Salaam City. A total of 286 Moran were interviewed. The results shows that 38.1% had no knowledge. 30.4% had low to moderate knowledge, and 33% had high knowledge about HIV/AIDS and its mode of transmission, prevention and voluntary counselling and testing (VCT). The level of knowledge about HIV/AIDS was associated with the participant's level of education. About 62% had negative attitudes toward preventive measures against HIV infection; and about 81% had never used condom in their lifetime. More than 75% were not aware of VCT. In conclusion, despite the fact that Tanzania's national average of HIV/AIDS knowledge is above 90%, there are isolated pockets in societies like the Wamaasai who require additional knowledge about HIV/AIDS transmission and methods of prevention. PMID- 20000015 TI - The impact of air pollution on hospital admission for respiratory and cardiovascular diseases in an oil-rich country. AB - OBJECTIVES: Aim of this study was to evaluate the impact of air pollution on hospital admissions for respiratory and cardiovascular diseases in an oil rich developing country, State of Qatar. METHODS: A prospective cohort population based study was conducted at different stations of Qatar during the period (2002 2005) for recording the concentration of air pollutants daily for sulphur dioxide (SO2), nitric oxide (NO), nitrogen dioxide (NO2), carbon monoxide (CO), ozone (O3) and particulate matter (PM10). Hospital admission data were collected from the inpatient discharge database of the Medical Records Department, Hamad General Hospital. RESULTS: An average of 5.36 admissions from ischemic heart diseases was counted daily in all the population which was even higher than the respiratory diseases (3.4/day). Minimum temperature was inversely correlated with all pollutants except for O3 and SO2. CONCLUSION: There was an association between increasing air pollutant levels and patients admitted for respiratory and cardiovascular diseases. PMID- 20000016 TI - Knowledge, attitude and practice of ante-natal attendees toward prevention of mother to child transmission (PMTCT) of HIV infection in a tertiary health facility, Northeast-Nigeria. AB - OBJECTIVE: Knowledge, attitude and practice of pregnant women toward prevention of mother to child transmission of HIV infection have not been evaluated in any tertiary health facility in the Northeast region of Nigeria. METHODS: A structured questionnaire was used to obtain data from 172 women that consecutively attended the antenatal clinic of University of Maiduguri Teaching Hospital to determine their level of knowledge, practice and attitude toward HIV/AIDS issues with respect to PMTCT. RESULTS: The survey revealed a high level of knowledge on modes of transmission, risks behaviours and prevention of HIV and other sexually transmitted infections among the pregnant women. However, the use of breast milk substitute (BMS) by HIV positive nursing mothers and use of condom during sexual intercourse did not receive very encouraging responses from 42 (24.4%) and 58 (33.7%) of the women respectively. Those that do not encourage use of BMS indicated spouse dislike as a major reason and the fact that the community places higher premium on breastfeeding than BMS. Majority of those that discouraged condom use 28 (48.3%), said the practice was against their religions beliefs while a few believes withdrawal before ejaculation and use of antibiotics after sex can equally prevent HIV infection. Majority of the respondents, 106 (61.6%) admit their willingness to support their spouses that tested positive for HIV. CONCLUSION: The pregnant women accepted PMTCT as a veritable means of preventing infants from HIV infection as well as an opportunity to know ones HIV status through voluntary testing. Majority implored greater involvement of their male partners and other significant family members during PMTCT counselling sessions to guard against ejection, stigma and discrimination if tested HIV positive. Staff training, awareness creation and community mobilization were identified as key to success of PMTCT programmes and fight against stigma and discrimination. PMID- 20000017 TI - Trends and determinants of under-five mortality in Uganda. AB - OBJECTIVES: We assessed the levels and trends of childhood mortality between 1990 and 2006, and assessed the determinants of under-five mortality. Results of this study are intended to aid formulate health policy interventions in the achievement of the Millennium Development Goals. METHODS: Three Uganda Demographic Health Survey (DHS) data sets collected in 1995, 2000/2001 and 2006 were used to estimate the levels and trends of childhood mortality between 1990 and 2006. The 2006 Uganda DHS was used to assess the determinants of under-five (0-59 months) mortality using a cross-sectional, nationally representative sample of 9,006 women aged 15-49 years who contributed 8369 children to the study. Proportions of the children were estimated, and Pearson's chi-square test was used to determine differences. Multivariate logistic regression analyses was used to assess the determinants of under-five mortality RESULTS: Under-five mortality remained unchanged between the periods 1991-1995 and 1996-2000, and then declined in the period 2001-2005. Results of the Cox regression modelling show that sex of the child (female) was associated with a lower risk of under-five mortality, while birth interval (less than 24 months after the preceding birth), type of birth (multiple), region of residence (northern region), and mother's education (primary, and no education) were associated with an increased risk of children dying before their fifth birthday. CONCLUSIONS: Uganda is not on track to meet the MDG Goal 4 (cutting under-five mortality by two-thirds by 2015-using the 1990 level as the baseline). However, with prompt multi-sectoral child health promotion strategies, coupled with diminished impacts of HIV/AIDS, and armed conflicts, improved maternal education, public enlightenment to lengthen birth intervals, improved living conditions in the north, the potential for Uganda to close the MDG 4 target gap remains high. Further studies are needed to assess the effects of contextual determinants of child survival in Uganda. PMID- 20000018 TI - Dental erosion due to lime consumption; review of literature and case report. AB - BACKGROUND: Dental health is increasingly threatened by dental erosion introduced by today's lifestyle. Extrinsic factor is the most implicated. Few literatures mentioned lime-incited dental erosion. CASE DESCRIPTION: A 49 year old woman was referred to our clinic with tooth wear and sensitivity. She was in good health but on daily dose of anti-hypertensive drugs for three years. Intraorally, classical erosive lesions were observed on the occlusal surfaces of the molars but of note are the lesions on the bucco-gingival surfaces of the maxillary incisors and the mandibular premolars. Dietary history eventually yielded inappropriate use of lime juice in order to slim down her weight. CLINICAL IMPLICATIONS: The use lime must be considered when assessing dietary causes of erosion. Extensive dental damage and dentine hypersensitivity are classical clinical features. Dentists must be able to make a diagnosis of lime-incited dental destruction and to institute the necessary therapeutic and preventive measures. PMID- 20000019 TI - Buruli ulcer in West Africa: strategies for early detection and treatment in the antibiotic era. AB - OBJECTIVE: Buruli ulcer (BU), caused by Mycobacterium ulcerans infection, has become one of the most rapidly emerging diseases in West Africa in recent decades. Until recently, the definitive treatment involved wide surgical excision. Recent data suggest that antibiotic therapy with rifampin and streptomycin may reduce the extent or prevent excision when initiated during the early phases of the disease. New strategies for BU control are needed, emphasizing early detection and increasing public awareness about the disease and treatment. Here we review current knowledge about BU and examine clinical, public health and anthropological research in the context of this new treatment paradigm to identify potential strategies for more effective control of this disease. METHODS: A comprehensive literature search for articles in English or French using the Medline and INIST databases, World Health Organization publications, and bibliographical references was undertaken using key words Buruli ulcer. Mycobacterium ulcerans, community surveillance, and public health. CONCLUSION: Studies to identify factors contributing to delayed presentation indicate that awareness of the disease is generally good in endemic regions, but wide variation exists in perceived cause of the disease, and the role of sorcery in its transmission and treatment. The use of traditional healers as first line therapy also contributes to delayed treatment, as do lack of awareness about the availability of effective treatment and financial concerns. Epidemiological data from existing BU control programs indicate that active public awareness campaigns are succcessful in increasing understanding and decreasing treatment delay and disease progression. Community-based surveillance and health education modeled after the village health worker programs used in the eradication of Guinea worm may be successfully applied in BU endemic areas. PMID- 20000020 TI - Application of the WHO five keys of food safety to improve food handling practices of food vendors in a poor resource community in Ghana. AB - AIM: To apply the WHO five keys of food safety in evidence based training programme for food vendors to improve the handling of street food. METHODS: A total of 127 food vendors in Accra, the capital city of Ghana were sampled for interviews. Data collection from the vendors focused on (i) food handling practices (ii) environmental and personal hygiene (iii) risk factors of oro faecal transmission. The data obtained from the study was analysed and utilised alongside the WHO Food Safety Training manual in a training workshop for the food vendors. Food handling practices of the vendors were re-assessed after the training workshop to assess the impact of the training. RESULTS: The main risk factor of oro-faecal transmission was the washing of food utensils with water that been used for hand washing after toilet attendance. Based on environmental and personal hygiene of the vendors, which were assessed by cleanliness, 16.5% and 4.7% respectively were scored as poor. Generally, majority of the food vendors practiced the WHO five keys of food safety, including keeping clean, separating raw and cooked food, cooking thoroughly, keeping food at safe temperature, and using safe water and raw materials. However, only a small proportion seems to practice them constantly. Impact assessment of the food safety training given to the vendors, showed that 60.5% of the vendors had acquired some knowledge from the workshop and were putting it into practice. CONCLUSION: Generally, food vendors have information on food safety such as hygiene and disease prevention. However, they require an impulse such as a training workshop to put knowledge on food safety into practice. PMID- 20000021 TI - Serological markers and risk factors for hepatitis B and hepatitis C viruses among students in a Nigerian university. AB - OBJECTIVE: The aim of this cross-sectional study was to estimate the seroprevalence rates of hepatitis B virus and hepatitis C virus infections and to analyze associated risk factors among 400 students in a Nigerian. METHODS: Participants were interviewed in a face-to-face sessions through a short structured questionnaire. Qualitative rapid immuno-chromatography test were used to screen for HBsAg and anti-HCV antibodies and these were confirmed by using third generation quantitative enzyme-linked immunosorbent assay (ELISAs). RESULTS: Seroprevalence rates were: HBV. All these individuals were asymptomatic and unaware of their hepatitis status prior to this study. We did not detect any case of HBV/HCV co-infection. The risk of being infected with HBV or HCV was not associated with marital status or having facial/tribal (p>0.05). A significant association was found between HCV (not HBV) infection and blood transfusion {OR=5.0 (95% CI=0.91-27.47); chi2=4.15; p=0.04}. CONCLUSION: Although the seroprevalence of HBV and HCV is lower among university students when compared to blood donors, HIV/AIDS patients and health workers in Nigeria. We have shown that these viruses circulate among the study population and blood transfusion identified as the factors most associated with HCV infection. Routine donor screening needs to be enforced and it is essential to implement prevention strategies focused on university students and youths in general. PMID- 20000022 TI - Associating gender with neighbourhood deprivation in Lagos State, Nigeria. AB - The study examines the effect of neighborhood deprivation on gender. It shows that most residents are deprived and that though, in Lagos State, the gender differences in the level of deprivation may not be too pronounced, yet, women, because of their socio-traditional roles in the society are more at a receiving end. There is gender difference in the distance an individual has to cover to get to the place of work with men having to cover more distance than women to get to their offices. The study shows that there is no difference between men and women in terms of accessibility to electricity and water as all residents are deprived almost equally. An interesting aspect of the study is that the fear of being killed or maimed as a result of crime within the neighborhood does not depend on the gender of the individual concerned. However, in general, there is gender difference in deprivation within the neighborhood with clear difference in the level of neighborhood convenience (chi2=97.131 and p<0.05) and availability of Amenities (chi2=105.12, p<0.05.) while there is no gender difference in neighborhood Safety (chi2=35.097, p>0.05), Health (chi2=63.933, p>0.05) and Community Relationship (chi2=12.905, p>0.05). PMID- 20000023 TI - Management and outcome of severely malnourished children admitted to Zewditu Memorial Hospital, Ethiopia. AB - OBJECTIVES: To describe the clinical profile and outcomes of severely malnourished cases admitted at Zewditu Memorial hospital, Ethiopia. METHODS: A retrospective descriptive data analysis of severely malnourished cases admitted to Zewditu Memorial Hospital from April 2005 to September 2008 was made. A total of 164 cases were enrolled and analyzed for various socio-demographic factors, comorbidities and outcomes. Both bivariate and multivariate models were performed to determine the outcome of the management by explanatory variables. Pearson's chi-square test of independence was used to test the existence of significant association of risk factors with the outcome. A p-value of less than 0.05 denoted significance in differences. RESULT: The predominant age group suffered from marasmus was the infants (75.4%) while kwashiorkor was prevalent during the second and third year and the difference noted was statistically significant. The mean age for marasmus, kwash and marasmickwash incidence was 16.9, 25.9 and 27.3 months respectively. The proportion of underweight was higher after the age of 60 months. Death occurred in 21.3% of the cases suggesting that mortality rate was higher than the acceptable range (21.3% vs. < 20.0%). Presence of diarrhoea (AOR=3.5, 95%CI=1.2 to 10.2), ocdema (AOR=0.2, 97%CI=0.1 to 0.9), stunting (AOR=3.3, 97%CI=1.2 to 8.2) and short mean duration of hospital stay (AOR=4.4 95%CI=2.0 to 10.1) were predictors of death outcome. CONCLUSION: The observed case fatality rate is unacceptably high and the risk factors for death are identified. In the face of many shortcomings in the hospital setting, managing uncomplicated cases of severe acute malnutrition is not encouraging when compared with the promising results of community based therapeutic care. We recommend the staffs to be trained and retained. PMID- 20000024 TI - Prevalence of cigarette smoking and the knowledge of its health implications among Nigerian soldiers. AB - OBJECTIVE: Several studies have reported a negative relationship between smoking and military performance. This study was conducted to determine the prevalence of cigarette smoking and knowledge of its health implications among Nigerian Army personnel. MATERIALS AND METHODS: A descriptive cross-sectional survey of 853 soldiers using a self-administered pre-tested semi-structured questionnaire. RESULTS: One hundred and seventy three respondents (20.3%) out of 853 respondents smoked. About three-quarter of them started smoking in the army. Derivation of pleasure and relaxation (24.9%), allaying anxiety (21.4%) and peer influence (34.1%) were the most frequent reasons for smoking. Skin disorder (75.9%), lung cancer (68.6%), addiction (65.2%) and dental problems (57.5%) were the most commonly reported effects of smoking. Only half (50.3%) of the total respondents believed that these effects of smoking could limit military fitness and performance. Forty five (26.1%) of the smokers had attempted to quit smoking. Majority (92.7%) of all the respondents have never had anti-smoking sensitization while in the army. CONCLUSION: The prevalence of smoking among the respondents was high. In spite of their knowledge its health hazards, the respondents could not relate this to military fitness and combat effectiveness. Also, their knowledge of the adverse effects did not translate to their smoking behaviour. There is need for continuous anti-smoking programmes to be established by the medical authority in the Nigerian Army to sensitize personnel on the dangers of cigarette smoking. PMID- 20000025 TI - Prevalence of gingivitis among school attendees in Qazvin, Iran. AB - OBJECTIVE: The purpose of this study was to determine the gingival status in six to nine years old (6-9 years). METHODS: A Total (n=792) patients comprised 396 girls and 396 boys between 6-9 years were selected from school in Qazvin. Names, ages, sex, addresses and gingival status of each patient were recorded in separate questionnaires. Intra oral examinations were performed with mirror, probe and good light. RESULTS: Among total n=792 patients, the female were 396 (50%) and male were 396 (50%). The prevalence of gingivitis was 97% (in 6-7 years), 98.1% (7-8 years), 98.5% (in 8-9 years) and 97.9% in all groups. CONCLUSION: In present study the frequency of gingivitis was found to be higher. According to the high prevalence of gingivitis the most prevalent areas of plaque and gingivitis identified in this study should be taken in consideration during oral hygiene instructions, which should be given to children, parents, and teachers. PMID- 20000026 TI - Determinants of condom use among antenatal clinic attendees in Dar es Salaam, Tanzania. AB - OBJECTIVE: To determine the demographic, socio-economic and psycho-social factors associated with condom use amongst antenatal clinic attendees in Dar es Salaam. METHODS: A cross sectional study design was employed in four antenatal clinics in Dar es Salaam. Pregnant women were interviewed between April 1995 and July 1997 to find out if they have ever used a condom and if so whether they had used them consistently for all coital acts in the previous year. RESULTS: Of 1,585 women interviewed, 41% had their first sexual experience before age of 18 years and 82% had a history of having more than two sexual partners during their lifetime. Sixty-two percent of women had never used a condom. Although 40% had used a condom in the previous year only 12% used them consistently. Ever use of a condom increased significantly with the number of years of education of the respondent and her partner also with the respondent's financial independence. Women with > 9 years of education were twice as likely as women with < 5 years of education to be condom users (prevalence ratio (PR) = 2.1, 95% confidence interval (CI) = 1.6 2.7). Professional women were almost twice as likely as housewives to have ever used a condom (PR = 1.8, 95% CI = 1.3-2.3). Women who reported that they have had more than four sexual partners during their lifetime were associated with nearly a four-fold higher lifetime rate of having ever used a condom, compared with a single lifetime partnership (PR = 3.9, 95% CI = 2.8-5.4). CONCLUSION: The reported prevalence of ever use of a condom amongst antenatal clinic attendees is low and inconsistent especially among HIV positive women. Deliberate effort should be used to ensure condom access, availability and correct and consistent use of condoms by women in all sexual acts. PMID- 20000028 TI - Community health seeking practices for the management of malaria of the under five in Bugiri District, Uganda. AB - OBJECTIVES: To obtain baseline information on health seeking practice of communities in management of the under-5 malaria fever that would indicate the outcome of introducing home management. METHODS: A cross-sectional study conducted in Bugiri District interviewing 451 heads of households at random throughout the district of, 411,250 population using structured and open questionnaires on health seeking practice in the management of malaria of under-5 in the communities. RESULTS: 20.2% of the communities took prompt action to treat children, although 51.7% consulted health workers (HW). Sources of treatment were drug shops (48.1%), and health units (42.4%), and very few herbalists. Few cases (7.8%) were referred to higher level. In the health units HW treated 65.7% of ill children with anti-malarial. At home caretakers gave anti-malarial to only 39.4%. Were communities to follow WHO diagnosis guidelines in home management, caretakers would treat 95.6% of illnesses with anti-malarial; an excess of 29.9% over HW. Large proportions (56.4%) of caretakers gave anti-malarial for treating any fever. 36.8% recognized Chloroquine as anti-malarial. Correct Chloroquine dose was used in 6.7%. Nearest health unit was within 2-3 km radius accessible on foot. Cost of management was affordable at Ugx 1,000/=. Referral units were far (25 km). CONCLUSION: Malaria is under-diagnosed, late and wrongly treated, with few referrals to higher levels. Home-based management will reduce morbidity and save more lives of malaria in under-5 but it will be like a mass treatment of children under-five in Bugiri District with the consequence of rapid introduction of drug resistance of the parasite, hence the need to improve on diagnosis to reduce unnecessary drug use. PMID- 20000027 TI - Associations between witnessing parental violence and experiencing symptoms of depression among college students. AB - OBJECTIVE: To examine the association between witnessing parental violence in childhood and experience of depressive symptoms during the academic year among college students in Awassa, Ethiopia. METHODS: A total of 2,708 undergraduate students (1,330 female and 1,378 male) completed a self-administered questionnaire that collected information concerning witnessing parental violence as a child, symptoms of depression during the current academic year. Logistic regression procedures were used to estimate odds ratios (OR) and 95% confidence intervals (95% CI). RESULTS: Approximately 22.7% female students and 27.1% of the male students reported witnessing parental violence. Females who witnessed parental violence were twice as likely to report moderate depression (OR=2.04:95% CI 1.32-3.15), or 3-times as likely to report moderately severe depression (OR=3.02:95% CI: 1.67-5.47) as compared with those who did not witness parental violence. The corresponding ORs were 1.71 (95% CI:1.13-2.59) and 2.42 (95% CI:1.41-4.13) for male students. Female students who witnessed parental violence were 2.4-times (OR=2.37, 95% CI: 1.26-4.44) more likely to report feeling bad about themselves 2.6 times (OR=2.62: 95% CI: 1.66-4.14) more likely to have trouble in concentrating compared with those who did not witness parental violence. Male students who witnessed parental violence were almost twice as likely to report having suicidal thoughts compared with their counterparts who did not witness parental violence (OR=1.97, 95% CI:1.16-3.34). CONCLUSION: Intervention programs focused on domestic violence must also address the needs of young adults from affected households. School-based counseling services may be one modality for addressing the needs of youths exposed to violence. PMID- 20000029 TI - Exposure to environmental tobacco smoke among adolescents in Kampala-Uganda, 2002. AB - OBJECTIVE: To estimate the prevalence of environmental tobacco smoke (ETS) inside or outside the home among school-going adolescents in Kampala, Uganda. METHODS: Data from the Kampala Global Youth Tobacco Survey (GYTS) of 2002 was used. We estimated frequencies and proportions of self reported exposure to ETS by the study participants. With logistic analysis, we assessed the association between ETS (outcome) in the home or outside the home and the following variables: sex; parental smoking status; and whether best friend was a smoker or not. RESULTS: Of the 2427 non-smoker teenagers who participated in this study, 52.8% were females, 17.9% were exposed to ETS at home while 48.7% were exposed to ETS outside of the home. The majority of the participants (65.8%) were in favour of banning smoking in public places such as in hotels, taxi, in schools, on playgrounds, in discos, markets, and shops. Compared to female participants, males were more likely to be exposed to ETS outside of the home (OR=1.21; 95% CI (1.02, 1.441). Having parents and close friends who smoked cigarettes was positively associated with exposure to ETS at home or outside of the home. Responders whose parents smoked cigarettes were more than four times likely to be exposed to ETS at home than those whose parents were non-smokers (OR=4.88; 95% CI [3.76, 6.33]). CONCLUSION: Cultural factors may expose boys to ETS than girls. We also found that having parents who were smokers exposed adolescents to ETS outside the home and having best friends who smoked exposed adolescents to ETS. This may suggest that adolescents who are exposed to ETS in one way may also be at risk of exposure through other means. PMID- 20000030 TI - Irrational antibiotic usage in boarding secondary school settings in Dar es Salaam. AB - OBJECTIVE: To assess the prevalence of antibiotic misuse among boarding secondary school students in Dares Salaam. METHODOLOGY: This descriptive cross-sectional study was conducted over 3-month period amongst boarding secondary school students. A randomized sample of students was recruited from at least 3 secondary schools from each of the three municipalities of Dar es Salaam region. Questionnaire with both closed-and open-ended questions was used for data collection in which socio-demographic characteristics, knowledge of ATBs, sources of prescription, procurement of ATBs, infections for which ATBs were used, and antibiotic therapy compliance were assessed. The collected data were analyzed using SPSS/PC+ version 15.0 computer package. RESULTS: A total of 424 randomly selected students with a mean age of 20 years were interviewed, of those 150 were females and 274 were males. Majority of the respondents (74%) have had heard of ATBs, however only 105 (25%) respondent described them correctly. Prevalence of antibiotic usage among the students was high (69%). Of 293 students who have had used ATBs, 73% of them had procured the drugs on prescription; 170 (58%) respondents used the ATBs for non-microbial infections/diseases, only 183 (62%) were compliant to antibiotic therapies (ABT), while 226 (77%) admitted to have had shared ATBs with friends/relatives whenever deemed necessary. Of those who had procured ATBs without prescriptions, 66% of them obtained the drugs from pharmacies. CONCLUSION: The study revealed inadequate knowledge on ATBs among the respondents and that pharmacies played a major role on easy availability of ATBs as over-the-counter (OTC) drugs attributing to high rate of irrational use of ATBs. We recommended that health care providers should succinctly explain to patients the importance of ABT compliance and adverse effects of non-compliance. Regulatory authorities should re-enforce the laws on sales of ATBs by prohibiting their availability as OTC drugs. PMID- 20000031 TI - Knowledge, attitudes and practices of local inhabitants about insecticide treated nets (ITNs) for malaria control in an endemic area of Ethiopia. AB - OBJECTIVE: The present study was conducted to assess the knowledge, attitudes and practices of local residents about insecticide treated nets (ITNs) for malaria control in an endemic area of Ethiopia. METHODS: A descriptive cross-survey was conducted from July 2008 to September 2008. A structured questionnaire collecting sociodemographic, ITNs and malaria-related KAP information was administered to the representatives of households. SETTINGS: Azendabo town, Ethiopia. RESULTS: 84% of the inhabitants had awareness about malaria transmission; mosquito bites were identified correctly as the cause of malaria. 85.2% respondents indicated that mosquitoes bite at night. ITNs awareness level of inhabitants was 92.4%; however, just 57.8% of them own ITNs. 96.2% and 91.5% of the inhabitants indicated the foremost reasons for nonpossession of ITNs were due to unaffordability and nonavailability. Inhabitants average monthly income significantly associated with the ITNs ownership at p<0.05 and, however educational status of the respondents and ITNs ownership was not significant (p=0.1). CONCLUSION: Indeed, respondents had adequate knowledge regarding ITNs and malaria related information. However, inhabitants had poor attitude and practice towards the ITNs utilization due to lack of availability and affordability. Therefore, besides health education campaign, distribution and promotion of ITNs to entire rural residents at free of charge may possibly bring constructive outcome in the near future. PMID- 20000032 TI - Mantoux skin test reactivity among household contacts of HIV-infected and HIV un infected patients with sputum smear positive TB in Dar es Salaam, Tanzania. AB - OBJECTIVE: To compare the prevalence of Tuberculosis (TB) infection as demonstrated by a positive Mantoux skin test (MST) among household contacts of sputum smear positive (SSP) HIV infected and un-infected TB patients. METHODS: A cross-sectional study of household contacts of patients with TB (index cases) attending Muhimbili National Hospital, and TB clinics of Infectious Disease Control Centre and Municipal hospitals in Dar es Salaam. Index cases with SSP-TB and aged > or =18 years had their HIV serostatus determined by ELISA. Contacts of index cases aged > or =1 year received intradermal protein-purified derivative (Mantoux test) and results were read after 72 hours. Infected contacts (MST positive) received medical evaluation to exclude TB disease. RESULTS: The overall prevalence of TB infection among contacts of HIV infected and un-infected index cases were 61.6% and 62.5% respectively, (OR 1.04 [95% CI 0.61-1.76], p=0.887). Factors associated with an increased risk of TB infection among contacts of both HIV infected and un-infected index cases were: older age of contacts (> or =25 years), duration of living with index case prior to TB diagnosis of > or =1 year, and sharing a room with index case. Infectiousness to their contacts was higher among female compared to male index cases. CONCLUSIONS: The proportion of household contacts of SSP PTB index cases with a positive MST was high and uninfluenced by the HIV serostatus of the index case. Efforts towards contact tracing and screening for TB among close contacts of SSP PTB patients should be enhanced. PMID- 20000033 TI - Anti-retroviral (ARV) rationing schemes in developing countries: a review article on strategies and ethical issues related to the successes and failures of ARV programmes. AB - When a patient receives a counterfeit drug, he/she becomes a victim of fraud medicine and is put at risk of developing adverse effects from unwanted medication that is not prescribed. These individuals health also becomes compromised because they are cheated of both successful treatment regimens and economically. Indeed counterfeit drugs pose many threats to society; not only to the individual in terms of the health side effects experienced, but also to the public in terms of trade relations, economic implications, and the effects on global pandemics. Apart from the pharmaceutical aspect in producing substandard drugs, there area also climatic or environmental factors as well as patients and economic factors. All these need to be addressed when considering any proper rationing strategy for antiretroviral drugs (ARVs) in sub-Saharan countries. Short of that, there is a great danger that the shelf life of ARV will be soon lost and once resistance comes in we will be in big problems. PMID- 20000034 TI - Female controlled methods of contraception and sexually transmitted infections including HIV in Malawi: examining the role of the female condom and microbicides. PMID- 20000035 TI - [The microglia activation characteristics of MA-induced neurotoxicity in the rats striatum]. AB - OBJECTIVE: To investigate the activation characteristics of microglia (MG) in the rats striatum with MA-induced neurotoxicity. METHODS: Male Wistar rats were divided randomly into control group (n=24) and experimental group (n=24). The rats of experimental group were injected intraperitoneally with MA (15 mg/kg x 8 injections, at 12 hours interval). The rats of control group were administrated with saline. The tissues of striatum of two rat groups were harvested at 0.5 d, 1 d, 2 d, 3 d, 4 d, 5 d, 6 d and 7 d post initial administrations of MA or saline. The structure changes were observed by transmission electron microscopy and CD 11b immunohistochemistry. The ratio of activated MG was calculated and statistically analyzed. RESULTS: In the control group, the morphological characteristics of the MG showed that the cell bodies were small with slender processes, high electronic density nucleus, and fewer organelles known as the "fork-type". In contrast, the MG in the MA-induced neurotoxicity group displayed larger cell body, shorter cell processes or disappeared, lower electronic density nucleus and rich organelles, resembling "bush-like" or "amoeba-like". The ratio of activated MG in control group was below 0.15 at all timepoints, whereas in the experimental group, the ratio of activated MG increased significantly from day 1 to day 7 (P<0.001). CONCLUSION: The continuous MA stimulation of the CNS results in prominent MG activation. PMID- 20000036 TI - [In vitro study on bFGF expression of rat astrocytes after mechanical injury]. AB - OBJECTIVE: To study the alteration of basic fibroblast growth factor (bFGB) expression in astrocytes in vitro after mechanical injury and to understand the repair mechanism of brain injury. METHODS: Astrocytes were isolated from cerebral cortex of SD rats born in 24 hours, and then cultured and purified. The cultured astrocytes were randomly divided into control group and injury groups that were subjected to mechanical injury at 30 min, 1h, 3h, 6h, 12h, 24h, 3d, and 7d. The levels of bFGF expression in the astrocytes after injury were detected by ABC immunohistochemistry. RESULTS: More than 95% of the cultured cells were astrocytes. The levels of bFGF expression werevery low in the control group. On the other hand, increased levels of bFGF expression could be observed at 1-3h after injury. The expression levels increased significantly at 6-12h, reached peak level at 24h, remained at the high level up to 3 days, and the decreased gradually. CONCLUSION: The changes of bFGF expression levels in cultured astrocytes in vitro after mechanical injury are similar to that observed in vivo experimental model, both of which show time-dependant characteristic, with only slightly earlier expression of bFGF observed in vitro. Thus, the expression of bFGF after injury can be one of evidences for estimation of brain injury intervals. the cell injury model in vitro may have superiority in the study of the molecule mechanism of tissue and cell injury. PMID- 20000037 TI - [The pathological observation of acute intoxication by Alangium Chinese in mice]. AB - OBJECTIVE: To investigate the pathological change of mice organ intoxicated by Alangium Chinese and its poisoning mechanism. METHODS: Mice were intoxicated by gavage with extract of Alangium Chinese. Then the histopathologic examination was made for evaluating the pathological changes in the organs of the poisoned mice by HE staining. RESULTS: The main pathological changes included alveolar hemorrhage, pulmonary interstitial hemorrhage, sinus hepaticus expansion and congestion, hepatocyte edema, subarachnoid hemorrhage, congestion and hemorrhage of other organs. CONCLUSION: The main target organs or tissue of Alangium Chinese are the lungs, liver and vascular smooth muscle. There is correlation between the toxic effect and the dosage. PMID- 20000038 TI - [The value of TNFalpha testing in evaluation of death due to drug anaphylactic shock]. AB - OBJECTIVE: To analyze the changes of the serum TNFalpha level in cadavers of drug anaphylactic shock(DAS) and to explore the significance of the TNFalpha testing in evaluation of death caused by DAS. METHODS: The level of serum TNFalpha from cardiac blood was determined by double antibody-sandwich ELISA method. RESULTS: The level of TNFalpha in the experimental group was (131.6+/-9.4) pg/mL, while the level in the infectious diseases control group and the normal control group were (87.3+/-6.4) pg/mL and (17.2+/-4.5) pg/mL, respectively. There was statistically significant difference among the groups (P<0.05). CONCLUSION: The serum TNFalpha level may be used as one of the indexes for evaluation of death caused by highly suspected drug anaphylactic shock. PMID- 20000039 TI - [Expression of monocyte chemoattractant protein-1 in sudden death due to viral myocarditis and its medicolegal significance]. AB - OBJECTIVE: To explore the expression and significance of monocyte chemoattractant protein-1 (MCP-1) in myocardium in sudden death caused by viral myocarditis (VMC). METHODS: To investigate the expression of MCP-1 in VMC sudden death group and control group using improved immunohistochemical technique and to compare the difference of the expression of MCP-1 between two groups with statistical method. RESULTS: In VMC sudden death group, MCP-1 was positively expressed in 17 of 20 cases. While only 4 of the 20 cases in the control group showed a mildly positive expression sparsely in myocardium, and the rest cases were completely negative. The rate of positive expression of MCP-1 in VMC group was obviously higher than that of the control group (P<0.01). CONCLUSION: The expression of MCP-1 detected by immunohistochemistry provides an objective morphologic evidence for the diagnosis of VMC sudden death in forensic pathology. PMID- 20000040 TI - [STR genotyping in unbuffered formalin fixed paraffin embedded tissue]. AB - OBJECTIVE: To assess the influential factors of STR genotyping in 10% unbuffered formalin fixed paraffin embedded samples. METHODS: Eight kinds of autopsy samples including heart, brain, liver, spleen, kidney, lung, stomach and intestine tissue from 2 corpse were fixed with 10% unbuffered formalin and embedded with paraffin according to the routine procedure from which the DNA were extracted with three different methods (QIAGEN, IQ and Chelex). STR profile were analyzed with AmpFlSTR Identifiler Kit and capillary electrophoresis on genetic analyzer 3100 Avant. STR profiles of 56 archival paraffin embedded samples from 15 cases were also analyzed with methods as mentioned. These archival samples, including heart, liver, lung and intestine tissue, had been preserved for 1 to 5 years in ambient temperature. Effectiveness of STR genotyping was assessed with the recalling ration of the 15 STR loci composing of the Identifiler Kit. RESULTS: Significant difference of the recalling ration was statistically revealed among the different types of paraffin embedded sample with same preserving period. Moreover, the STR recalling ration was continuously lowering with the prolongation of preserving period in all of the samples. The linear relationship between the STR recalling ratio and the preserving period was showed in lung and heart sample. The STR recalling ration in lung sample was higher than that in the other types of paraffin embedded sample. CONCLUSION: Preserving period, tissue type, extracting method of DNA and the PCR template concentration were the most important influential factors for successfully STR genotyping paraffin embedded samples, which fixed with unbuffered formalin for the same time. PMID- 20000041 TI - [Full sibling identification based on the number of matched STR locus]. AB - OBJECTIVE: To establish and evaluate the method of matched locus numbers in full sibling identification. METHODS: Two hundred and eighty full sibling (FS) pairs and 2003 unrelated individual (UI) pairs were genotyped with Identifiler system. The number of locus matched with 0 identical allele (A0), matched with 1 allele (A1) or matched with 2 alleles (A2) were counted and full sibling index (FSI) were calculated based on ITO method. Fisher discriminant functions were established based on the numbers of matched STR locus or FSI. Power of different Fisher discriminant functions was statistically analyzed. RESULTS: The distribution of A1 and A2 in FS group and that of A0 and A1 in UI group were in accord with normal distribution. Contrarily, A0 in FS group and A2 in UI group fitted to skew distribution, respectively. Difference of A1 distribution was not statistically significant in the two groups (P>0.01). The established Fisher discriminant functions based on A0 and A2 for each group were Z(FS)=0.99817A0+4.24442A2-12.77970 and Z(UI)=2.01456A0+1.54658A2-7.28076, respectively, the average error probability of which was as low as 0.0490. The power of discrimination for full sibling showed no statistically significant difference between ITO method and the established Fisher discriminant functions. CONCLUSION: The number of matched STR locus in Identifiler system is a valuable method in full sibling identification. The discriminating power of the established Fisher discriminant functions based on the matched STR locus number is similar with that of classic ITO method in full sibling identification. PMID- 20000042 TI - [Polymorphic distribution of FUT2/01 in northern Han Chinese population]. AB - OBJECTIVE: To investigate the sequence features of FUT2/01 locus and its polymorphic distribution in Chinese population, and to discuss its application potential in forensic medicine. METHODS: The alleles on FUT2/01 locus were amplified by PCR and then were sequenced. Furthermore, polymorphic distribution of the locus was analyzed by polyacrylamide gel electrophoresis. The genotypes were characterized with fluorescence labeling followed by automatic detection system. RESULTS: The sequencing results only showed the length differences which were determined by the tandem repeats variance of the core sequence. There were 9 alleles and 28 genotypes identified from 162 individuals. The discrimination power and excluding probability of paternity were 0.9639 and 0.6266, respectively. In addition, the locus could be genotyped by automatic analysis very well. CONCLUSION: The FUT2/01 locus exhibits high heterozygosity and individual identification power in Chinese Han population, and may be a valuable STR system for application in forensic medicine. PMID- 20000043 TI - [The correlation between ketamine-induced schizophrenia-like signs in mice and the expressions of NRG1, ErbB4 mRNA]. AB - OBJECTIVE: To explore the correlation between signs similar to schizophrenia in mice after ketamine administration and the expressions of NRG1 and ErbB4 mRNA in order to explain the possible pathogenesis of schizophrenia. METHODS: Fifty KM mice were randomly divided into 5 groups which were administered intraperitoneally with saline, clozapine and different dosages ketamine. The ketamine groups were administered intraperitoneally with low dosage (25 mg/kg), middle dosage (50 mg/kg) and high dosage (100 mg/kg) one time every day for 7 days. After administration of 100 mg/kg ketamine for 7 days, the clozapine group was introgastrically administered 20 mg/kg with clozapine one time every day for 7 days. The pathological changes of hippocampus neurons were observed by HE stain. The expressions of the NRG1 and ErbB4 mRNA in hippocampus were detected by reverse transcriptase polymerase chain reaction (RT-PCR). RESULTS: In the group with high dosage of ketamine, the levels of NRG1 and ErbB4 mRNA were significantly lower than that of the group with saline. CONCLUSION: Ketamine may induce signs similar to schizophrenia in KM mice. The mechanism may be involved in the reduction of NRG1 and ErbB4 mRNA expression. PMID- 20000044 TI - [The application of diminished criminal responsibility rating scale to mental retardation offenders]. AB - OBJECTIVE: To explore the application of Diminished Criminal Responsibility Rating Scale (DCRRS) to mental retardation offenders. METHODS: The DCRRS was used to 121 cases of mental retardation offenders who were divided into three groups according to the degree of their diminished criminal responsibility. RESULTS: There were significant differences in rating score among the three groups (mild group 22.12+/-4.69, moderate group 25.50+/-5.48, major group 27.59+/-5.69), and 17 items had good correlation with the total score of the scale with the correlation coefficient from 0.289 to 0.665. Six factors were extracted by the factor analysis, and 69.392% variation could be explained. CONCLUSION: The DCRRS has rational items, its total score could show the difference among the three degree diminished criminal responsibility of mental retardation offenders. PMID- 20000045 TI - [Effect of expanding training on prisoners' mental health]. AB - OBJECTIVE: To study the effect of expanding training on the mental health of the prisoners and to provide the references to the innovation of the mental health education for the prisoners. METHODS: Questionnaire, observation, interview and mathematical statistic analysis were used. 100 male and 100 female prisoners from two prisons in East China were investigated by Symptom Checklist 90 (SCL-90). RESULTS: Eight of the ten factors in SCL-90 showed the level of statistically significance, which includes obsessive-compulsive symptoms, interpersonal sensitivity, depression, anxiety, compulsion, hostility, psychoticism and others. Expanding training could be helpful to improve the mental health of the prisoners. CONCLUSION: Expanding training can be used in mental health education of the prisoners. PMID- 20000046 TI - [Stability of the tetramine in blood samples under different temperatures]. AB - OBJECTIVE: To study the stability of tetramine in blood samples under different environment temperatures. METHODS: The tetramine samples in 0.5 microg/mL concentration were stored in the environment temperature at 45 degrees C, 25 degrees C and 4 degrees C. The tetramine concentration were measured by GC-MS in 1, 3, 12, 18 and 39 days, respectively. And then the results of measurement were analyzed by SPSS software. RESULTS: At 45 degrees C and 25 degrees C, the concentration of tetramine changed without distinctly in first 3 days and declined significantly from 4 days to 12 days. Thereafter it declined more slowly. At 4 degrees C, the concentration of tetramine changed without distinctly in 12 days and declined significantly from 13 days to 18 days. Thereafter it declined more slowly. CONCLUSION: The concentration of tetra mine in blood samples is stable in 3 days. Thereafter it declined gradually. Temperature also influences the concentration of tetra mine. PMID- 20000047 TI - [Analysis of 324 cases of forensic psychiatry expert testimony in Chongqing]. AB - OBJECTIVE: To study the current situation and features of forensic psychiatric identification in Chongqing. METHODS: The demographic data and criminological characteristics of 324 cases were analyzed using self-made questionnaire. RESULTS: There were 322 cases in which opinions to diagnosis and law-related items were both given. One hundred and forty-five cases were schizophrenias (45.03%), 51 were mental retardation (15.84%), 78 were others (24.22%), and 48 showed no psychosis (14.91%). In legal conclusion, 211 cases (65.53%) were evaluated for criminal responsibility and 41 (12.65%) were evaluated for competence of sex defense. CONCLUSION: Most cases has psychosis shows the importance and necessity of forensic psychiatric identification, and various jurisprudence conclusions indicated the diversity and complexity of these cases. PMID- 20000048 TI - [Forty-five cases on re-assessing the personal disability]. AB - OBJECTIVE: To discuss the reasons for the alterations of the conclusions in re assessment for personal disability in road traffic accidents, work-related accidents and other accidents. METHODS: Forty five cases of re-assessing the personal disability from 2007 to 2009 in our institute were collected and analyzed. RESULTS: The disability grade of 12 cases (26.7%) was altered in re assessment. The reasons for the alternation included assessment standard items misquoted, assessment time mischosen, injury misdiagnosed and omitted, artificial prostheses mismanaged, injury-disease relationship erroneous judgment and disability grade promoted incorrectly. CONCLUSION: The attention for assessment of disability grade should been paid to correct time for assessment, correct items of identification standard, appropriate medical and forensic technology, and correct understand about all relevant regulations. PMID- 20000049 TI - [Sixty cases with eyes injuries degree in forensic medicine]. AB - OBJECTIVE: To discuss the characteristics and main problems of eye injuries in forensic identification. METHODS: Sixty cases of eye injury in forensic expertise were retrospectively analyzed according to sex, age, employment, trauma-causing instrument and injury type, respectively. RESULTS: Of the 60 cases there are 61.7% being peasants and workers, 85.0% suffering from blunt trauma and 63.3% suffering from simple contusion. CONCLUSION: Eyes injuries was mostly caused by blunt trauma. Pathological change of fundus was an important factor affecting the vision function. The injury-causing instruments, injury sites and medical history of eyes should be considered while evaluating the vision. PMID- 20000050 TI - [Analysis of 112 cases with diffuse axonal injury]. AB - OBJECTIVE: To investigate the relationship between the diffuse axonal injury (DAI) and cerebral contusion, primary brain stem injury and brain concussion. METHODS: One hundred and twelve cases with DAI were analyzed according to the characteristics of clinical signs and imaging features. RESULTS: Of 112 cases of DAI, 70.5% injured in traffic accident, 60.7% injured with blunt trauma more than one time and 71.4% injured with cerebral contusion. And 90 cases with brain with hemorrhage were found in CT or MRI imaging. CONCLUSION: DAI may be associated with cortical contusion and primary brain stem injury. The CT or MRI is useful to investigate the cause of death and to evaluate the personal disability. PMID- 20000051 TI - [Progress in skeletal age estimation with measurement method of X-ray film]. AB - Age estimation by the measurement method is to get the data such as the length, width, thickness, area of the ossification center, transverse diameter of the metaphysis, transverse diameter of osteoepiphysis, width of the epiphyseal line and the ratio between the diaph and the epiphyseal, by measuring the different bones of human in different ages with the radiological technology and to explore the relationship between these data and the life age by statistics. According to the relationships the personal life age may be estimated. In this article the measurement methods above mentioned and its application in age estimation were reviewed. PMID- 20000052 TI - [DNA-based personal identification in disaster]. AB - The disaster is a sudden unexpected event that causes serious human injuries and deaths as well as missing persons. The main tasks of forensic DNA laboratories are to identify victim in the disaster. After reviewed the previous disasters and related studies, we proposed a new procedure of DNA identification for the use of disaster in the future, which includes preparation works, samples collection, samples storage, DNA extraction, typing, data analysis and interpretation of results. Some experiences and problems about the DNA identification are also discussed. PMID- 20000053 TI - [Psychological autopsy and its limitation in application]. AB - The suicide were analyzed depending only on the death scene and cadaver features but not on the psychological autopsy (PA) that also enables the determination of the manner of death in China for a long time. There are significant differences between PA and the traditional risk assessment of suicide. The goals of PA are to find out the features of individuality, the patterns of behavior and the possible motives of suicide. The methods of PA involve investigating, collecting and analyzing all related information of the deceased. There are many limitations of PA in application mainly due to the lack of the standard procedures. Although an improved procedure has recently developed by Berman, many problems need to be solved before the actual application of PA in China. PMID- 20000054 TI - Equity and utilization of preventive health care services. The case of immunization completion among children 12-23 months in Kagera region Tanzania. AB - OBJECTIVE: The aim of this study was to determine factors associated with completion of immunization as an input in to strategies to raise the immunization coverage in Tanzania. METHODS: This study used a panel data collected over four survey rounds at a minimum of six months apart to determine the causes of failure to complete immunization in Tanzania. The data were collected during between 1991 and 1994 in Kagera region Tanzania among children 12-23 months old. Data analysis was done using Statistical Analysis Software (SAS) version 9.1. Pooled logistic regression was used to determine the likelihood odds ratio of completing immunization. RESULTS: The study observed 550 children contributing 1541 children observation rounds across all panels. Immunization coverage was 87.7 percent and factors that significantly (p<0.05) increasing completion of immunization were having more than three under five children, high mother's education, being urban, road passable through out the year and high economic status. If head of household was a female compared to male it significantly reduced the odds of completing immunization. CONCLUSIONS: Strategies to improve immunization coverage have to give priority to households headed by a female, poor and those with poor access to health care. PMID- 20000055 TI - Blood lead levels and potental environmental exposures among children under five years in Kibera slums, Nairobi. AB - OBJECTIVE: Lead exposure has been associated with intellectual impairment in children in a number of international studies. Prevalence of elevated blood lead levels (eBLL > or = 10ug/dL) of between 5 - 15% has been reported among in Nairobi (UNEP, 2006). However, little is known about potential environmental exposure for eBLLs among children in Kibera, Nairobi. METHODS: A descriptive, cross-sectional study of children drawn from Kibera slums who presented at Yes to kids (Y2K) programme of VIPS Health Services at Woodley, Nairobi between June and August 2007 was carried out. The study assessed potential correlates of eBLLs in 387 children aged 6 to 59 months and had lived in Kibera slums since birth. Sampling was purposive. The factors examined were age, sex, breastfeeding history, respondent's education and occupation, type of house walls, sources of drinking water and kales, and awareness of lead poisoning among respondents. Potential risk factors such exposure to paint, contaminated playgrounds, glazed pottery, cosmetics and para-occupational as well as living near lead industry and pica behavior were also examined. Potential environmental sources of lead such as drinking water, soil and kales were analyzed for lead levels. RESULTS: Seven percent (n = 27, N = 387) had BLLs above 10ug/dl. BLL > or = 10ug/dl was associated with non-permanent housing (p = 0.812), playing on potentially lead contaminated grounds (p = 0.627) and pica behavior (p = 0.439). Low risk parental occupation (p = 0.001) and Kales sourced from the market/kiosks (p = 0.001) were significantly associated with BLL > or = 10ug/dl. Soil lead levels (Soil Pb) ranged from 3,000 to 90,000ug/kg, which was very high compared to WHO acceptable range of 100 - 200ug/kg. There was weak linear association (r2 = 0.0160) between Soil Pb and mean BLLs for a given village. There were no detectable levels of lead in kales and tap water. CONCLUSIONS: The study found about 7% (N = 387) of the children tested had eBLL > or = 10ug/dl in an area with very high soil lead levels (range in Kibera slums: 3,365 - 89,570ug/kg; WHO allowable range: 100 - 120ug/kg), raising a health flag that must be addressed using the multi-sectoral approach and further studies. It's important to note that the study design and its inherent limitations could have masked true picture of childhood lead poisoning in Kibera slums, Nairobi. PMID- 20000056 TI - Maternal age at birth and under-5 mortality in Nigeria. AB - OBJECTIVE: The paper examined the impact of maternal age at birth on under-5 death in Nigeria. METHOD: Secondary data were generated from the 2003 Nigeria Demographic and Health Surveys in examining the relationship between maternal age at birth and under-5 mortality risk. Relationships between variables were tested through bivariate and logistic analyses. RESULTS: Out of 7620 sampled women for the study, almost 60% were less than 30 years old, the median age was 26 years- a youthful population. Analyses of the data revealed a high under-5 mortality rate (45.4%), a general high home delivery (62.4%) among Nigerian women, which dictates a low rate at which assistance by health professional is being sought during childbirth . Furthermore, mothers' median age at first birth was less than 19, while under-5 death was significantly pronounced among younger (less than 20 years) mothers and older women (above 35 years) (p<0.05). Maternal education which was significantly low among younger mothers was a predictor of under-5 mortality. CONCLUSION: Under -5 mortality is still high especially among younger mothers. Thus resolving this challenge in Nigeria will be inadequate if early childbearing issues are not addressed using a tailored framework alongside with the need to improve maternal education in Nigeria. PMID- 20000057 TI - Diagnosis of malaria by community health workers in Nigeria. AB - OBJECTIVE: The introduction of primary health care made Nigeria, a developing country, train and retrain community health workers to work all over the country especially in the rural communities where there is dearth of doctors. Despite their training and experience many people are skeptical of their competence to diagnose accurately what more treating endemic disease like malaria. The need to find out the diagnostic competence of the health workers in malaria control programme now in Nigeria necessitated this study. METHOD: A rural primary health centre, the sentinel site for malarial control programme investigation in Imo State of Nigeria was selected. The community health technician (CHT) was the health worker in charge. Those who were diagnosed as malaria patients by CHT were examined by a medical laboratory scientist (who was engaged specifically for this job) for malaria parasitaemia. The laboratory examination was Giamsa - stained thick blood from fingerprint. Those with positive parasite density count at 1000/microL and above were regarded as malaria patients. The study was from March - October 2007. RESULTS: The number diagnosed as malaria patients on clinical grounds by CHT was 2512 while the number diagnosed by both clinical and laboratory basis was 2490. The number of patients with wrong diagnosis of malaria by CHT was 22 (0.875%). CONCLUSION: The CHT is useful in the diagnosis and by extension in the control of such endemic disease as malaria where there is no laboratory facilities. Both the employers and populace should repose confidence in their services and in the area where they have been trained and acquired experience PMID- 20000058 TI - HIV seropositivity and CD4 T-lymphocyte counts among infants and children in Port Harcourt, Nigeria. AB - BACKGROUND: Most published reports on HIV prevalence are centered on adults of sexually active age and antenatal mothers with little or no regard to infants and children. METHODS: A total of one thousand (1000) children aged 1 to less than 15 years attending the various clinics at the Braithwait Memorial Specialist Hospital (BMSH), Port Harcourt were recruited into the study. Blood samples (3 millilitres of blood) were collected into a five millilitres syringe and the serum was used for serological testing of antibodies to human immunodeficiency virus (HIV) and CD4 cell counts from EDTA blood. The WHO HIV testing strategy III was adopted for defining the HIV status. The CD4 cell count was estimated using the Dynamal Quant Kit (Dynal Biotechn, ASA, Oslo, Norway). RESULTS: The overall HIV prevalence rate in this study was 23.2%. The distribution of HIV prevalence among different age group revealed a high prevalence rate among the under fives (24.1% for males and 26.4% for females). There was a significant negative correlation between age and HIV status of children (r = -0.94, P <0.01). The mean CD4 count of the infected children was 647.25 +/- 340.1 (95% CI 603.3 - 691.1). Logistic regression analysis showed that both age and sex exert an influence of the CD4 count only in the under fives (0 - <5 yrs). 10.3% of the children were immuno compromised with CD4 count below 200 cells, 27.9% fell into the moderate category while 61.8% had normal CD4 counts of 500 cells and above. CONCLUSION: The seroprevalence of HIV among infants and children is high in this part of the world and this call for urgent intervention through voluntary counseling of antenatal mothers and proper monitoring in order to reduce the paediatric HIV prevalence. PMID- 20000059 TI - Prevalence and risk factors of low back pain among nurses in Africa: Nigerian and Ethiopian specialized hospitals survey study. AB - BACKGROUND: The mechanical hazards in the hospitals include low back pain (LBP) from manual lifting (lifting patients in particular) which makes nursing one of the occupations most affected by LBP. Nurses are required to lift and transport patients or equipments, often in difficult environment particularly in developing nations like Africa, where lifting aids are not always available or practicable. The objective of this study was to determine the prevalence and risk factors of LBP among nurses in African. METHODS: A cross-sectional study was designed and used to determine the prevalence and risk factors for LBP among nurses in a typical Nigerian (Murtala Muhammed Specialist Hospital [MMSHI) and Ethiopian (Jimma University Specialized Hospital [JUSH]) Specialized Hospitals. A department-to-department enquiry was conducted using a self structured valid and reliable questionnaire. Simple percentage (%) and Chi square were used to analyze variables of interest. RESULTS: Five hundred and eight respondents (178 [35%] males and 330 [65%] females) participated in the study. The 12 month prevalence of low back pain (LBP) was 360 (70.87%). LBP was more prevalent among female nurses (67.5%) than the male nurses (32.5%). It was also associated with occupational hazard and poor knowledge of back care ergonomics. The prevalence of LBP was highest among nurses in Obstetrics and Gynecology Unit (26.67%) and least among tutors (4.17%).There was no significant difference between Nigeria and Ethiopian nurses' responses in prevalence, etiology and knowledge of back care. However, there was a significant association between gender, knowledge of back care ergonomics and prevalence of LBP at p< 0.05. The prevalence of LBP at MMSH (Nigeria) and JUSH (Ethiopia) is comparable to levels recorded outside Africa. However, in this study LBP did not feature as a major cause of sickness absence in the work place contrary to those reported outside Africa. Nurses only lost 202 days (0.15%) of the total working (131,400) days, this is considerably very low. Though, longer sick leaves (7563 days) were medically advised and applied for, However, only about 2.7% of the applied sick leaves was granted on technical or/and administrative grounds against medical recommendation and advices. CONCLUSION: It was concluded that poor back care ergonomics, duty stress and unavailability of lifting equipments are the major predisposing factors of LBP among nurses in Africa. RECOMMENDATION: It was recommended that regular refresher courses on back care ergonomics are essential. PMID- 20000060 TI - Tetanus toxoid vaccination coverage and differential between urban and rural areas of Bangladesh. AB - BACKGROUND: Government commitment and support from a range of partnerships have led to a massive increase in tetanus toxoid immunization coverage among women of childbearing age, ensuring that both mothers and babies are protected against tetanus infection in Bangladesh. In order to control and eliminate the vaccine preventable diseases it is important to know the vaccination coverage. OBJECTIVES: The major objective of this study is to determine the complete vaccination rate and the predictors that influence vaccination of mothers during pregnancy and to see whether there is any gap lies between the women of urban and rural areas regarding the tetanus toxoid injection receiving. METHODOLOGY: This study utilizes the data extracted from Bangladesh Demographic and Health Survey 2004 (BDHS).To meets the objectives this study considers bivariate and multivariate analysis. RESULTS: The study represents that 88 per cent urban mothers and 84 per cent rural mothers receive tetanus toxoid injection during their pregnancy period. Logistic regression model is adjusted by wealth index, mother's age at last birth, education, husband's occupation, ever using contraception, fertility preference, wanted last child, having permission to go to hospital/health center, telling about pregnancy complications and mass media exposure for receiving TT injection. All these explanatory variables come out to be as significant determinants of receiving TT injection for all mothers as well as for rural mothers in Bangladesh. On the other hand ever using contraception, wanted last child, telling about pregnancy complications, mass media exposure and wealth index are the significant determinants of receiving TT injection for mothers of urban area. PMID- 20000061 TI - Physical activity and dietary fat as determinants of body mass index in a cross sectional corelational design. AB - Overweight/obesity and related disease conditions will constitute a major threat to the economically productive adults and subsequently, will present a huge health-care burden on developing countries in the near future. Suspected determinants include physical activity and dietary fat. The main indicator of overweight/obesity is Body Mass Index (BMI . The purpose of this article is to present the prediction power of physical activity and dietary fat intake on BMI of lecturers within a higher learning institutionalized setting. The studyadopted a cross-sectional correlational design. Proportionate and simple random sampling techniques were used to select a sample of 120 lecturers who participated in the study. Data collection was conducted through questionnaires, which had sections including physical activity checklist, 24-hour food recall, anthropometrics measurements mainly weight and height. Analysis of data involved the use of bivariate correlation and linear regression. A significant inverse association occurred between BMI and minutes spent in moderate intense physical activity per day (r=-0322, p<0.01). Physical activity also predicted BMI (R2=0.096, F=13.616, beta=-3.22, t=-3.69, N=120, P<0.01). However, the association between Body Mass Index and dietary fat was not significant (r=0.038, p>0.05). In conclusion, physical activity was a significant predictor to BMI and on the contrary no significant impact was caused by dietary fat intake. Therefore, we still need further investigations on the effect of physical activity and dietary fat on BMI and risk factors associated poor diet should take priority. PMID- 20000062 TI - Cultural factors and home-brewed alcohol use in a rural community in South Africa. AB - This paper presents an analysis of the relationship between cultural orientation and home- brewed alcohol use among adolescents in a rural setting. The study involved 1263 students from 30 high schools in Mankweng District in Limpopo Province of South Africa. Boys formed 48% of the sample. Means ages were 16.7 and 16.0 years for boys and girls respectively. A 16 item scale focusing on language, mass media and music preferences was constructed to measure cultural orientations. There was an integration of western and traditional/local activities by respondents who reported having used home-brewed alcohol in the past 12 months. In some respects the sample had western tendencies i.e. English newspapers were read more than no-English newspapers and funky music tended to be preferred to traditional South African music, while in others the reverse was true i.e. Radio Thobela (a radio station that uses vernacular languages) was listened to more than Radio Metro (a radio station that uses English language only). There were significant differences between boys and girls in the use of western media. Older adolescents (18 years or older) used more home brewed alcohol than the younger ones. The concept of culture and its associations with alcohol use invites a critical view on cultural values among adolescents. The positive association with home-brewed alcohol use among secondary school students in our sample has been demonstrated for an orientation towards external cultural influence. Educators and learners need to increase their consciousness on the problematic side of cultural import. PMID- 20000063 TI - Attitude of self ear cleaning in black Africans: any benefit? AB - Wax removal compromises the integrity of the ear's defenses. It is a leading cause of otitis externa, Otomycosis and impaired hearing. Aims of the study are to assess the knowledge and implication of self ear cleaning among black Africans. MATERIAL AND METHODS: A prospective study carried out at the Tundun wada community and National Ear Care Centre, Kaduna with administration of structured questionnaire after an informed consent and ethical clearance. Information retrieved includes biodata, believe, reason and object for cleaning of the ear, associated problems or benefit and examination. RESULTS: A total of 372 form the basis for the study, age range 1-76 yr with a mean age of 30.37 yr, median age 29.00 (S.D. = 13.79) and M:F ratio of 1:1. Mother responded for their children except for grown up. About 47.3% of the subjects were unemployed who were either children or complete house wife. About 90% of the subjects interviewed do self ear cleaning and over 90% believe ear should be cleaned to remove wax, because of itching in over 50% while a few is due to cosmetic reason. Cotton bud was the commonest material used for cleaning. About one-third of the subject has formed the habit unconsciously over 10 years. The entire subjects interviewed had their ear examined; about 27% had ear discharge then wax impaction in 22% then foreign body 12%. CONCLUSION: The habit of self ear cleaning should be discouraged as it is slow otologic poison with an attendant long term effect. PMID- 20000064 TI - Awareness regarding communicable diseases among the patients attending general out-patient department of a tertiary care hospital of Kolkata, India. AB - OBJECTIVE: Communicable diseases continue to be a major problem in India, a large number of which can be prevented by health education, targeting specially the deficiencies and for which a baseline assessment is essential. METHODS: This observational study included OPD patients who were selected by random sampling and interviewed. The age range varied from 20 - 50 years. RESULTS: 52.5% resided in rural areas. 80% were literates. Majority had per capita income below Rs 1000 per month. Awareness regarding symptoms, modes of transmission, prevention and management of the studied diseases were not satisfactory. Majority used apparently safe water. Mosquito-net use was more among literates. Media was the main source of information in urban areas, health functionaries in rural areas while relatives, friends and neighbours influenced the residents of both the areas and the finding was statistically significant. CONCLUSIONS: An awareness development programme involving a combination of different health education methods is suggested. PMID- 20000065 TI - Bacteriological evaluation of pre-cut fruits sold in Kano metropolis, Kano State, Nigeria. AB - One hundred and fifty (150) pre-cut fruit samples comprising of Pineapples (50), Paw-paw (50) and Watermelon (50) at the point of stand retail outlets were tested by standard bacteriological methods to determine bacterial contamination of the fruits. Out of these 150 examined 136 (90.67%) were contaminated with bacteria. The bacterial distribution were; Escherichia coli 69 (46.00%), Staphylococcus aureus 29 (19.33%), Salmonella species 13 (8.67%), Proteus species 18 (12.00%), Enterobacter aerogenes 3 (2.00%), Klebsiella pneumoniae 2 (1.33%) and Pseudomonas aeruginosa 2 (1.33%). Among the 50 Pineapple pre-cuts, Escherichia coli 26 (17.33%), Staphylococcus aureus 6 (4.00%), Salmonella species 7 (4.67%), Proteus species 9 (6.00%), Pseudomonas aeruginosa 2 (1.33%); the 50 Watermelon had Escherchia coli 22 (14.67%), Staphylococcus aureus 13 (8.67%), Salmonella species 3 (2.00%), Proteus species 5 (3.33%), Enterobacter aerogenes 2 (1.33%), Klebsiella species 2 (1.33%). Of the 50 Paw-paw pre-cuts were: Escherichia coli 21 (14.00%), Staphylococcus aureus 10 (6.67%), Salmonella species 3 (2.00%), Proteus species 4 (2.67%), Enterobacter aerogenes 1 (0.67%) were isolated. The findings in this study have shown that the food vendors failed to adopt adequate hygiene for food handling and thus, suggest that the quality of all the pre-cut fruits sold at the retail outlets were not bacteriologically satisfactory. The public health risks associated with these pre-cut fruits may suggest that these fruits could serve as the vehicles for foodborne illnesses. This study has shown the need to educate the vendors on how to protect utensils and fruits to avoid contamination and spoilage. PMID- 20000066 TI - Effect of joint iron and zinc supplementation on malarial infection and anaemia. AB - OBJECTIVE: In spite of interventions being put in place to reduce the burden of maternal anaemia and malaria in Ghana, they continue to impact negatively on pregnancy outcomes. It is unclear whether there are some other factors that limit the effectiveness of these interventions. The aim of this study was to explore whether there are benefits associated with the addition of zinc to the routine malaria chemoprophylaxis, iron and folic acid intervention package for pregnant women in Ghana. DESIGN: A double-blind, randomised controlled trial (RCT) conducted in the Upper West Region of Ghana. RESULTS: Iron-zinc supplementation reduced the risk of increased malaria parasitaemia associated with high maternal serum ferritin and Hb concentrations. Iron-zinc supplementation was associated with reduced malaria parasite densities, F (1, 20) = 4.744, p = 0.042. Iron and zinc provided in the ratio of 1:1 prevented a significant decline in maternal iron stores of pregnant women. Adjusted geometric mean serum ferritin concentration in the Iron-zinc Group was significantly higher than in the Control Group (22.9 microg/L versus 16.9 microg/L), F (1, 156)= 6.336, p = 0.013 CONCLUSIONS: Joint iron and zinc supplementation appears to be a better option than iron-only supplementation in malaria-endemic areas. PMID- 20000067 TI - The burden of malaria infection on pregnant women and birth weight of infants in south western Nigeria. AB - OBJECTIVE: To determine the effect of malaria infection on pregnant women and the birth weight of the infants in the south western Nigeria. SUBJECT: 262 pregnant women who came for antenatal clinic at Ade-Oyo maternity hospital. 128 were primigravidae while 134 were multigravidae. METHODOLOGY: 2ml of blood was withdrawn from 262 pregnant women who came for antenatal clinic at Ade Oyo maternity hospital. Thick blood smears were prepared for parasite identification and quantification. Anaemia was detected by measuring Hb levels using Drabkin's solution. Age, gravidity and history of treatment with antimalaria drugs were obtained from the subjects using questionnaire. RESULT: The overall prevalence of infection was 41.8%. Primigravidae were more infected (35%) than multigravidae (22%). The prevalence was significantly higher (p<0.05) in wet season than dry season. Teenagers and primigravidae were more infected than the adults and multigravidae. The severity of the anaemia was significantly higher (p<0.05) among malaria positive teenagers and primigravidae than adults and multigravidae. The mean birth weight of infants born to malaria positive was significantly lower (p<0.05) than those born to malaria negative mothers. Malaria positive teenagers and primigravidae had infants with lowest birth weight as compared with adult and multigravidae. The birth weights of the infants were positively correlated with the Hb levels. CONCLUSION: This study suggests that malaria infection, anaemia, and gravidity affect the birth weight of infants born in Ibadan, southwestern Nigeria. PMID- 20000068 TI - Barriers to exclusive breastfeeding among infants aged 0-6 months in Eldoret municipality, Kenya. AB - OBJECTIVE: Breast milk is the best source of nutrients for young infants. It promotes optimal growth and development. WHO recommends Exclusive Breastfeeding (EBF) for the first 6 months of life as the best way of feeding an infant. EBF Prevalence in Kenya is low, there is need to find out barriers to EBF, hence the study. The objective of the study was to establish barriers to EBF to 6 months among infants aged 0-6 months. METHODS: The study was cross-sectional involving 384 mother-infant pair visiting Huruma and West, maternal and child health (MCH) clinics. This study was done in the urban setting of Eldoret. Simple random sampling technique was used to get the desired sample size of 384. A questionnaire was used to collect data. Data was analyzed using Statistical Package for Social Scientist (SPSS) version 12.0. RESULTS: Maternal age was categorized into mothers with; <24 years (n=207, 53.9%), 25-29 years (n=116, 30.2%), 30-35 years (n=35, 9.1%) and >35 years (n=24, 6.3%). Over half (n=198, 51.5%) of infants were female and 48.5%( n= 186) were male. The median and mean age of infants in the study was 2.3 and 2 months respectively. Reported barriers to exclusive breastfeeding included; breast milk unsatisfying to the infant (n=157, 64.4%), insufficient breast milk production (n=35, 14.3%), to improve nutritional status of the infant (n=18, 7.4%), for the infant learns to feed on other foods (n=19, 7.7%) and in order for mothers to resume work (n=15, 6.1%). CONCLUSION: Barriers to EBF in the study may be attributed to inadequate breastfeeding knowledge among the mothers. This study is important in devising strategies that will increase EBF in the community. PMID- 20000069 TI - Knowledge of HIV/AIDS and preventive measures use by city commuter-bus drivers and conductors in Dar Es Salaam, Tanzania. AB - OBJECTIVE: The study was aimed at assessing the knowledge of the drivers and conductors on HIV/AIDS epidemic and its preventive measures. METHODS: This descriptive study was prospectively conducted among randomly selected city commuter-bus drivers and conductors from December 2006 to April 2007 in Dar Es Salaam, Tanzania. The study parameters included the general knowledge of the respondents on HIV/AIDS, treatment by anti-retroviral therapy and different types of preventive measures. Data was collected through face-to-face interviews using semi- structured questionnaires. RESULTS: A total of 333 respondents voluntarily participated in the study. It was found that out of all respondents who had heard about HIV/AIDS, 93.9% heard it from the media particularly radios, televisions and newspapers. The results further showed no significant difference (p=0.08) of knowledge on HIV/AIDS between the drivers and conductors. Drivers (84.3%) and conductors (80.1%) pointed out condom as the common method of prevention from contracting HIV/AIDS. Among other preventive measures known to them, 75.5% and 52.9% of the drivers and conductors pointed out faithfulness respectively. However results indicated that 49.30% of the respondents were practicing faithfulness to their partners as a preventive measure while 40.9% were using condoms and only 9.6% were practicing abstinence. CONCLUSION: This study has shown that city commuter-bus drivers and conductors in Dar Es Salaam, have "adequate" knowledge on HIV/AIDS and preventive measures and the major source of this knowledge is through the public media. PMID- 20000070 TI - Biodemographic and health seeking behaviour factors influencing neonatal and postneonatal mortality in Bangladesh: evidence from DHS data. AB - This study findings show primarily - amongst the biodemographic and health seeking services factors, delivery-related maternal health complicacies, blindness, higher order births, twin births, lower household size and interaction effect of higher order live births and male child are significantly correlated with higher neonatal mortality. Neonatal deaths are heavily caused by biological, demographical and maternal experience health hazards during/after delivery. The analysis shows that the causes of deaths after neonatal period are deeply rooted in poverty, regional administrative disparity, lack of breastfeeding, unplanned frequency of births, small interval between births and non-utilization of health seeking services. Education, even maternal, and sex differential have no significant effect as what the literature suggested. But the interaction effect of maternal secondary and above education who residing in urban areas has a negative significant association with neonatal mortality. Increased interval between the births significantly reduced the postneonatal but not the neonatal mortality whereas the relationship between the child's birth order and neonatal is found significantly positive. It is suggested that increasing the length of births interval and the duration of breastfeed lowering the frequency of births should decrease the risk of neonatal and postneonatal mortality. Nutrition factor breastfeeding is negatively associated with postneonatal mortality; as duration of breastfeeding increase the postneonate deaths decrease. Results show that the interaction variable of higher order births and the child is boy has moderately significant positive association with neonatal mortality. Postneonates residing in Sylhet have exceptionally higher likelihood of mortality. Although credit for contributing to the lowering of infant mortality has been given to health programs by public health personnel and to the improvement in socio-economic status by social scientists, but in Bangladesh both of these factors are found to influence early childhood mortality. Maternal and child health program as well as public health and nutrition program should be strengthened to provide modern healthcare services to all pregnant women and their infants. PMID- 20000071 TI - Prevalence and correlates of mental distress among regular undergraduate students of Hawassa University: a cross sectional survey. AB - INTRODUCTION: Mental health is gradually being recognized as an important public health concern. There is high level of mental distress in the community. Student integration in Universities is usually difficult and involves many factors which are potentially stressing. OBJECTIVE: To assess the prevalence of mental distress (Common mental disorders) among regular undergraduate students of Hawassa University, and to study its correlates METHOD: This cross sectional study was conducted among regular undergraduate students of Hawassa University, during the period May 28-June 6, 2007, which was after the mid semester exam in the second semester of the academic year September, 2006- July, 2007. 'SRQ-20 items' was used to measure the prevalence of common mental disorders among the students. A total of 1198 students were included in the study. RESULTS: There was very high level of mental distress (49.1%). Highest levels of mental distress were observed among students of the Main Campus, in the Faculty of Business and economics, among freshman students and those students who never follow religious programs irrespective of their religion. Difficulty in making friends and dating; active sexual practice; conflicts in the dormitories with fellow students; income and stationary materials inadequacy; lack of adequate access to: academic reference materials; lack of adequate access to sanitary facility and recreational facility; overcrowding and worrying about personal safety were associated with higher levels of mental distress. CONCLUSIONS AND RECOMMENDATIONS: Detailed and focused study about the campus facilities that were proved to be deficient; correction of the deficiencies and the inclusion of a course in the freshman students' curricula that focuses on the mental well being of the young adult, means of coping with stress, possible challenges in the university life and other related issues (with their solutions) are discussed. PMID- 20000072 TI - The health and living conditions of children in child-headed households in Siteki, Swaziland. AB - OBJECTIVE: The focus of the study reported on here was to determine the health and living conditions of children living in child-headed households (CHHs) in Swaziland, where it is estimated that approximately 10-15 percent of the entire population will be orphans and other vulnerable children (OVC) by 2010 and that one in ten households in the country today are child-headed because of the impact of HIV and AIDS. METHODS: The population comprised 41 heads of household, caring for 97 siblings among them. Data was collected in a single period between the months of February and April 2007 using the convenience sampling method and employing a semi-structured questionnaire as the data-collection instrument. RESULTS: Children were generally physically healthy, despite not receiving adequate food or balanced diets. Most had access to health facilities or at least to sources of medication. Education assistance exists but is limited and abuse was generally not reported for fear of reprisal. Children generally relied on family and community networks for assistance, but the strain this put on those offering assistance meant that the assistance was not always consistent. Some children also relied on NGOs for food donations, but many of the children were unaware of the services offered by these organisations. CONCLUSIONS: There is a dearth of information on adolescents in Swaziland, the primary caregivers in most child-headed households. Children are not actively consulted and encouraged to participate in helping to solve the problems they are affected by. There is a lack of coordination between caregiver organisations, leading to ineffective and inefficient service provision for this particular vulnerable group. PMID- 20000073 TI - Micronutrient profile of children and women in rural Bangladesh: study on available data for iron and vitamin A supplementation. AB - BACKGROUND: Micronutrient deficiencies are so important to public health outcomes, particularly in the developing world, that a series of global goals have been established, and significant amounts of donor and national funds have been directed at them. OBJECTIVES: This paper makes an attempt to compile the micronutrient profile of children and mothers (available data on vitamin A and iron supplementation) in rural Bangladesh and to identify the factors influencing with respect to these micronutrients intake. METHODOLOGY: To reach our goal Bangladesh Demographic and Health Survey of 2004 data for last five years (N = 3329) has been used. Descriptive and multivariate logistic regression methods were employed in analyzing the data. RESULTS: It is observed that only 64% of targeted children had received vitamin A dose in the six months preceding the survey and overall, 14% and 45% mothers received a postpartum vitamin A dose and iron tablets. The study elucidates that the rate of receiving vitamin A dose by children lower in Barisal division than among other five divisions. Inequalities were observed in receiving vitamin A and iron supplementation, regarding household quality and assets index. The proportion of receiving vitamin A dose by children and intake of iron by mothers was found higher living in the upper quality houses. Middle aged mothers more received iron tablet and vitamin A dose and less sufferer form from difficulty with night blindness and day light vision during pregnancy than among the adolescence and older aged mothers. Multivariate logistic regression analysis considering mothers vitamin "A" dose and iron supplementation as a dependent variable shows that higher educated women were approximately two times higher probability of receiving vitamin "A" supplement of their child than women with no education. Women whose husbands had a lower status of job were less likely to receive vitamin A dose and iron supplements. The other main contributing factors likely to affect micronutrients intake among women were mass media, mother's age at last birth, mother's and husband's education and husband occupation. CONCLUSIONS: The results indicate several policy options: (a) there is need to ensure availability of iron and vitamin a supplements; (b) there is also the need for creation of awareness regarding micronutrient intake among mothers and children through programmes like mass media campaign; (d) The high risk group such as adolescents and higher aged women need special care and the existing health management system may be strengthened to create awareness among mothers of these groups for micronutrient intake from the beginning of pregnancy; (c) it is equally important that education for women and increased cash incomes is emphasized to bring about a lasting impact on the overall nutrition status of women and children. PMID- 20000074 TI - Oral hygiene and periodontal status of hemodialyzed patients with chronic renal failure in Qazvin, Iran. AB - AIM AND OBJECTIVE: The primary purpose of this investigation was to evaluate the oral health and gingival status of individual's undergoing renal diialysis in Qazvin province. METHODS AND MATERIALS: One hundred eighty Iranian patients on hemodialysis participated in the study, and were divided into three subgroups of the population were studies: 1) those who have been on renal dialysis for less than one year; 2) those on renal dialysis for 1 to 3 years; and 3) those on renal dialysis for longer than 3 years. Four dental indices: the debris index (DI); the calculus index (CI); the plaque index (PI) and the gingival index (GI) were used. Data were compiled and analyzed by using 1-way analysis of variance (ANOVA). RESULTS: The results of study showed a 100% (n=180) of the individuals undergoing renal dialysis presented prevalence of mild to moderate gingivitis among the studied population. The mean of debris index were 1.65, 2.07, and 2.15, with SD +/- 0.67, 0.47 and 0.48 respectively for the subgroups. The means of plaque index were 1.72, 2.16 and 2.26, with SD +/- 0.64, 036 and 0.42 respectively for the revealed groups. The means of calculus index were 1.58, 2.02 and 2.09, with SD +/ 0.58, 0.28 and 0.39 respectively for the subgroups. The means of gingival index were 1.43, 2.97 and 2.06 with SD +/- 0.67, 0.38 and 0.35 for the subgroups respectively. Turkey's post hoc test showed significant difference in all indices between the 1st and 2nd subgroups, and between the 1st and 3rd subgroups, while no significant difference was found between 2nd and 3rd subgroups. CONCLUSION: It was concluded that periodontal it was concluded that periodontal disease is prevalent in renal dialysis patients who showed unacceptable level of oral hygiene and which may increase with the chronicity of the illness. Findings led to the conclusion that the renal dialysis population in Qazvin Province, regardless of length of time on dialysis, is in need of comprehensive professional oral care and self-care instruction. Oral disease was present and is a source of active infection in these medically compromised individuals and, as such, has dire implication for morbidity and mortality. PMID- 20000075 TI - [Analysis of reasons for late recognition of tuberculosis and cancer of the lung and the ways of enhancing the efficiency of their differential diagnosis]. AB - Difficulties in the differential diagnosis of tuberculosis and cancer of the lung were analyzed in 272 patients from a regional tuberculosis dispensary. The efficiency of using the real-time polymerase chain reaction (PCR) to detect Mycobacterium tuberculosis DNA molecule fragments in the sputum specimens from 111 patients of this category was studied to improve the differential diagnosis of these diseases. The real-time PCR was ascertained to be a rather highly sensitive (78.3%) and highly specific (96.1%) technique for tuberculosis diagnosis, which considerably improves the etiological recognition of the pathological process. Examinations of patients with peripheral lung cancer and benign round shadow formations in the lung yielded encouraging results. The specificity of the technique in this pathology is as high as 100%. PMID- 20000076 TI - [Extrapulmonary tuberculosis according to the data of personal registries of new tuberculosis cases]. AB - By using the subject registers of new tuberculosis cases, which are kept in the subjects of the Russian Federation within tuberculosis epidemiological monitoring, the authors analyze notified extrapulmonary tuberculosis morbidity in 21 subjects (17,301 patients over 1995-2006). Despite the stability of the pattern of recorded pulmonary tuberculosis (PT) as a whole, great differences are found in the notification rate of different forms of PT between individual subjects of the Russian Federation, there are wide differences in the proportion and absolute number of PT patients first detected in the coterminous subjects of the Russian Federation with similar natural climatic and socioeconomic conditions. The registered patients with PT are shown to be a heterogeneous group of patients who substantially differ in gender, age, and the forms of disease therefore the use of total parameters to assess the situation, by detecting and diagnosing PL, is not efficient. PMID- 20000077 TI - [New trends in the epidemic situation of extrathoracic tuberculosis in Siberia and the Far East]. AB - The statistical reports by 21 subjects of the Russian Federation (the Siberian and Far-Eastern Federal Districts) have been analyzed to define the rates of extrathoracic tuberculosis morbidity in these regions and its structure. It has been established that new trends have been recently evident in the pattern of extrathoraric tuberculosis. Firstly, there has been an increase in the incidence of bone and joint tuberculosis, young men and old women being predominant in its structure. Secondly, the steep rise in the incidence of genital tuberculosis, caused by the appearance of an experienced phthisiological gynecologist in one of the regions emphasizes that the fact that there is a large proportion of undetected extrathoracic tuberculosis cases is pressing. There is concern for the high percentage of children with extrapulmonary tuberculosis--this fact reflects the tense epidemic situation in the areas. PMID- 20000078 TI - [Main reasons for a need for retreatment of pulmonary tuberculosis in Armenia in the past 10 years]. AB - 250 patients in need of retreatment for pulmonary tuberculosis were enrolled in the study. The later indicated that there was a preponderance of males constituting all-bodied age population groups aged 25 to 64 years in the age and gender structure of the examinees. The major factors contributing to reactivation of the tuberculous process and poor outcomes of a previous course of specific therapy were extensive X-ray changes in the lung, drug resistance of the pathogens of tuberculosis to first-line drugs, and neglect of X-ray findings in the determinants of the duration of treatment. The use of the standard short-term course of chemotherapy was found to result in incomplete cure of disseminated and moderately disseminated processes, particularly in drug resistance of Mycobacterium tuberculosis and in concomitant diseases, which becomes a cause of early and late recurrences. PMID- 20000079 TI - [Adjuvant lymphotrophic chemotherapy in the surgical treatment of progressive pulmonary tuberculosis]. AB - The study was undertaken to enhance the efficiency of surgical treatment for progressive drug-resistant pulmonary tuberculosis, by applying adjuvant lymphotropic chemotherapy (ALCT). Seventy-one patients were examined and operated on; of them 35 patients received ALCT and 36 patients formed a control group. The determinants of the efficiency of the performed treatment were the frequency of specific postoperative complications. ALCT used in combination with the conventional antituberculous treatments made it possible to enhance the efficiency of surgical treatment, by halving the number of postoperative complications, reducing the duration of a postoperative period, and enhancing the intensity of chemotherapy, by taking into account the data on the resistance of a pathogen, without causing adverse reactions, and to improve the psychoemotional status of patients. PMID- 20000080 TI - [Comparative data on the use of high performance liquid chromatography to identify mycobacteria isolated on liquid and solid media]. AB - Two hundred and forty-two mycobacterial cultures isolated from clinical materials on the Lwenstein-Jensen solid medium and 208 cultures in the automatic Bactec MGIT 960 system (on the modified Middlebrook 7H9 liquid medium) were examined. Of them, there was M. tuberculosis (85 and 82, respectively), M. kansasii (26 and 24), MAC (46 and 38), M. xenopi (24 and 20), M. fortuitum (26 and 22), and M. chelonae/abscessus complex (20 and 18). Identification of mycobacterial cultures isolated on the solid medium by microbiological assay and high performance liquid chromatography (HPLC) showed the agreement of results of the cultures isolated on the liquid medium in 95.5 and 97.2%, respectively; that by these techniques revealed rapidly growing nontuberculosis mycobacteria in 95.8 and 95.2% of cases, respectively; slowing growing mycobacteria in 91.7 and 97.8%, and M. tuberculosis in 96.5 and 97.6%. Mycobacterial isolation on the Middlebrook 7H9 liquid medium in the automatic Bactec MGIT-960 system takes a shorter time than that on the solid (Lwenstein-Jensen) medium. The microbiological identification of mycobacteria lasts as long as 3 weeks while the use of HPLC reduces its time to 24 hours. The efficiency of HPCL does not depend on whether mycobacterial cultures are isolated on the solid or liquid media. PMID- 20000081 TI - [Species-specific detection of Mycobacterium tuberculosis complex]. AB - The routine polymerase chain reaction (PCR) was used for the species-specific detection of related Mycobacterium tuberculosis complex (MTC) strains characterized by a high level of nucleotide sequence homology. Nucleotide sequences for the 16S rRNA, rpoB, gyrB genes of MTC strains, which were potential markers for their genotyping, were analyzed. The constructed phylogenetic trees confirmed the accuracy of the current taxonomic classification of MTC. Five sets of primers were developed for the species-specific detection of the mycobacteria M. tuberculosis, M. canettii, M. microti, M. bovis, and M. caprae. It has been shown that PCR by means of the primers having oscillating 3'-terminal nucleotides (complementary DNA of only a certain mycobacterial type) may be used to genotype MTC in the correct choice of the temperature of primer annealing. PMID- 20000082 TI - [Equipping the bacteriological laboratories of a tuberculosis service as the most important element of assurance of the quality and safety of their work]. AB - By taking into account the present requirements for the equipping of the laboratories, the authors have drawn up the minimum standard list of equipment, as well as a list of additional equipment for the specialized bacteriological laboratory of a tuberculosis-controlling institution, which performs microbiological studies for the diagnosis and control of chemotherapy for tuberculosis. The specifications and characteristics of the baric types of equipment used to fit out the laboratories under the present conditions are described. Equipping the laboratories in accordance with the draw-up lists is shown to ensure a qualitative, effective, and safe work. Recommendations on how to supply the laboratories with equipment, to make the optimal choice, and to use consumables for tests are given. PMID- 20000083 TI - [Use of automated bactec MGIT 960 system in the diagnosis of resistance to reserve drugs agents in Samara]. AB - The paper gives the results of a demonstration project to introduce a test for drug sensitivity to second-line antituberculous agents for patients with multidrug-resistant (MDR) tuberculosis at a routine large Russian laboratory. Two hundred and thirty MDR isolates were examined; of them 8.7% had extreme drug resistance. The cost of this test was estimated to be US $ 33.4. The introduction of this test is possible and essential for the precise and timely choice of antituberculous therapy for patients with MDR tuberculosis. PMID- 20000084 TI - [Physical factors of the urban dwelling environment: ecological and hygienic aspects]. AB - The paper describes the ecological and hygienic significance of the physical factors of the urban dwelling environment: the macroclimate of a residential area, the microclimate within the residential and public buildings, the ionic ozone regimen of premises, an electromagnetic load on the population, the problems associated with noise and light exposure. It also gives an integrated assessment of the current air-conditioning systems, household electric appliances, and building engineering systems. A system of measures is proposed to optimize the quality of the urban dwelling environment. PMID- 20000085 TI - [Reactions of different cell populations of rats to intake of waters treated with physical methods]. AB - A 6-month experiment was carried out to study the reactions of different cell populations of the testicle and duodenum of rats and the gastrointestinal tract of volunteers to the intake of 3 waters treated by physical methods: water conditioned with electric pulsed discharges; 2 waters obtained using energoinformational technologies: Grander water (Austria) and immunomodulated water Renorm (Russia). The test waters treated by physical methods were shown to have no negative effect or a positive effect on animals and man. Three used procedures to evaluate different gastrointestinal tract portions (oral and intestinal mucosae) of rats and volunteers revealed the preservation of the mucosal epithelium when Grander and Renorm waters are taken in doses. PMID- 20000086 TI - [Temperature dependence of the structurized state of water]. AB - The paper provides the results of studies of the impact of temperature on the structural and energetic state of drinking water and shows that the altered temperature conditions of water causes a change in its supramolecular structure and biological value. PMID- 20000087 TI - [The location of wind-mill electric generating plants: hygienic aspects]. AB - The hygienic aspects of the location of wind-mill electric generating plants become more pressing due to the intensive development of wind-power engineering. Possible risk factors from wind-mill electric generating plants that can influence the environment and the population are considered. A 400-m control area is recommended on the basis of the made calculations of an acoustic and electromagnetic pollution area, an exposure area during emergency situations, as well the field measuring data on noise from wind-mill electric generating plants with a capacity of 20 MW, by applying 100-kW wind-mill electric generating units. Further studies are proposed to improve the differentiated sizes of control areas for wind-mill electric generating plants with wind-mill electric generating units of varying capacity. PMID- 20000088 TI - [A cause of the abnormal properties of melt water]. AB - The physicochemical properties and crystal structure of melt water were studied. Melt water was found to be in the active metastable state with the non equilibrium concentration of hydrogen and hydroxyl. The biological activity of such water is accounted for by its small sizes of supramolecular complexes. Films of structural formations were first obtained and their acoustic emission in the homogeneous medium recorded. PMID- 20000089 TI - [Integrated assessment of the environment in an area of location of radiation dangerous objects]. AB - The authors proposed to use control levels depending on the category of an area to make an integrated hygienic and ecological assessment of the state of the area in the exploitation of radiation-dangerous objects as a main criterion for radiation and chemical factors. The average data of recent ecological monitoring on the level of radionuclides and chemical elements in all environmental objects (soil, atmospheric aerosols, surface water, and bottom sediments) may be used as background indicators in this area. The background indices and double mean-square deviation were proposed to take as a basis for calculating the control levels. The proposed model for calculating the control levels may be used to assess the state of an area of any radiation-dangerous objects. PMID- 20000090 TI - [The soil content of cesium-137 in the Republic of Altai due to nuclear tests at the Semipalatinsk testing site]. AB - Radioactive fall-outs from the atmospheric nuclear weapon tests carried out at the Semipalatinsk testing site have resulted in local soil pollution with 137Cs in the Republic of Altai. The subjects of the investigations were the main types of soils from North and Central Altai. In the Republic of Altai, the share of 137Cs in the upper (0-5 cm) soil layer has been established to be 54.6% in the Turochaksky district to 92.1% in the Ulagansky district. By taking into account the fact that the major activity of 137Cs has been accumulated in the 0.5-cm surface layer, its may be expected that there will be its rather separation with a fine-dispersed surface material In this connection, 137Cs can enter the organism with inhaled dust due to deflation during soil eolation. The inhalation of 137Cs by the dwellers of the areas that have been exposed to irradiation during nuclear weapon tests at the Semipalatinsk testing site continues to be also important today. PMID- 20000091 TI - [Study of the consequences to the health of the Urkutsk Region's population from nuclear explosions at the Semipalatinsk testing site]. AB - Analysis of archival records on the activity of diurnal plane-tables from the region's weather stations revealed local radioactive fall-out in the near-Baikal areas from the nuclear weapon tests carried out at the Semipalatinsk testing site. Examination of mortality rates in the settlements exposed to the tests showed that the maximum rates of overall and lung malignancy mortalities were observed in 1960-1979; the mean radiation dose in the exposed settlements were estimated to be 580-850 MeV. PMID- 20000092 TI - [The need for human health protection from climate changes]. AB - The occurring climate changes have a considerable impact on human health. The higher frequency of different natural disasters, including flood and typhoons, deteriorates the epidemiological situation. Evidence is provided for the importance of climate warming as a risk factor for a number of communicable diseases, including West Nile fever in the Astrakhan and Volgograd Regions. The paper presents the results of analytical epidemiological surveys, by using the time-series analysis, in Tver and Moscow, which indicate an increase in the number of fatal outcomes particularly among the elderly during temperature heat and cold waves. The specific features of the impact of climate warming on human health in the northern and southern regions of the country are considered. The need for developing the National Plan of Actions to prevent the population's health from climate changes is warranted. PMID- 20000093 TI - [Hygienic assessment of priority physical factors of the urban dwelling environment]. AB - A complex of chemical, physical, and biological factors of varying intensity influences man under the dwelling environmental conditions. Modern man spends as high as 80-90% of the day time under the conditions of the closed spaces of the urbanized habitat and is constantly exposed to physical factors. The present environmental hygiene monitoring system requires great attention to the control of priority physical factors in the environment. By using a number of examples, the authors give criteria for determining the priority of factors of physical nature and an objective assessment of their importance in the formation of the safe dwelling environment. PMID- 20000094 TI - [Shielding of the geomagnetic field in apartment houses]. AB - The present investigation has studied the specific features of shielding of the natural geomagnetic field (GMF) in the multistorey apartment houses. The results of the experimental study of the shielding properties of some building materials are presented. It is shown that there are 1.2 to 2.0-fold reductions in GMF induction within the houses depending on the number of storeys and the properties of building materials. The level of GMF within the multistorey apartment house decreases by an average of 27.2+/-0.5 microTl from the 1st to the 9th floor and the GMF attenuation factor in many apartment houses exceeds the permissible limit for working places, especially on the top floors. On designing new building materials and constructions and on architectural design, one should keep in mind their shielding properties. PMID- 20000095 TI - [Scientific bases for the methodology of complex prediction of the impact of global hydroclimatic changes on the medical and ecological environment]. AB - Global hydroclimatic changes present a threat to human health in the areas where there may be a drastic intensification of negative (direct and indirect) influences of a water factor. The developing poor climatic situation requires that the prediction theory be improved to have an effective apparatus for substantiating the managerial decisions aimed at protecting the population's health in the changing the human habitat. The paper describes scientific approaches to the prognostic patterning of such kind. PMID- 20000096 TI - [Impact of the structuredness of water on the formation of its biological activity against leukocyte phagocytic activity and cell-mediated immunity]. AB - The effects of waters of varying structuredness on the body's immunological responsiveness were investigated. Long-term intake of highly-structured waters was shown to lead to the stimulation of delayed hypersensitivity reaction and the activation of leukocyte phagocytic activity. PMID- 20000097 TI - [Preclinical health changes upon exposure to hydrocarbons of petroleum genesis]. AB - A full-scale toxicological experiment established a complex of biochemical, clinical, and immunological parameters for the diagnosis of preconditions upon exposure to hydrocarbons, which was tested on children from two towns differing in the degree of ambient air pollution owing to hydrocarbons. PMID- 20000098 TI - [Physical factors of the industrial environment and traffic safety]. AB - The paper deals with the problem of optimization of working conditions of engine drivers in the control cabs of up-to-date locomotives, by toughening the sanitary and-hygienic requirements for the levels of bad industrial factors. The authors associate the optimization of working conditions with the necessity of strengthening the reliability of their driving activity due to the increasing rail movement speeds and the transfer to one-person rolling-stock operation, and other factors. PMID- 20000099 TI - [Evaluation of the influence of physical environmental factors on morbidity among armed forces personnel]. AB - Under the conditions of hot and mountain-continental climate, the morbidity rates were estimated to be significantly lower than those in young men who had not been acclimatized or adapted to living conditions and in non-acclimatized men. A role of individual physical environmental factors (temperature, relative humidity, barometric pressure, average and maximum air speed) and integral exposure by the wind chill index (a combined impact of an air speed and ambient temperature) as risk factors to human health was defined. The mountain-continental climate showed a relationship of the influence of these factors to habitation at different altitudes. PMID- 20000101 TI - [Retrospective view of the use of detoxifying hemosorption for the treatment of acute exogenous poisoning]. AB - Results of the treatment of almost 5000 patients with severe forms of acute poisoning by hemosorption (HS) using non-selective hemosorbents is a basic method comprising specific and non-specific therapeutic modalities that facilitate removal of toxicants of different nature from the human body. It is shown that natural biotransformation of toxicants and correction of disturbed hemostasis can be accelerated by a combination of HS and physico-chemical hemotherapy (magnetic, UV, and laser therapy, indirect electrochemical blood oxidation by sodium hypochlorite) that promotes further significant increase in the toxicant removal rate. Improvement of detoxification therapy by combining HS with other afferent methods for detoxification and physico-chemical hemotherapy allowed the mortality rate in the department of emergency antitoxic therapy to be reduced by a factor of 3, from 14.5 to 5.8%. PMID- 20000100 TI - [Evaluation of the efficiency of operation of MFM-0142 membrane module-based units in the sanitary-virological monitoring of water]. AB - The paper outlines materials on how to improve a method for indication of water viral pollution in different water objects of the environment, by applying a MFM filtration module based on tangentially radial-mode water microfiltration and viral concentration on the positively charged membranes, by using the secondary stage of concentration. Experimental and full-scale studies have demonstrated that this method for virus isolation has a high effectiveness and is based on the employment of the materials made in Russia. This determines its rapid introduction into the system for the sanitary-virological monitoring of practical laboratories of various departments, which will favor the higher efficiency of epidemiological surveillance as an integral part of nonspecific prophylactic measures of virus infections of water etiology. PMID- 20000102 TI - [Extracorporeal liquor-corrective therapy of traumatic subarachnoidal hemorrhage]. AB - Comprehensive analysis of criteria for the severity of clinical conditions in patients with craniocerebral injuries included characteristics of cerebrospinal fluid and lesions of hematoencephalic barrier (HEB). The study revealed the relationship between clinico-neurological conditions of the patients, liquor nitrite levels, and HEB permeability. An attempt was undertaken to evaluate efficiency of liquor-corrective treatment in combination with intense therapy in patients with severe cerebral injuries. PMID- 20000103 TI - [Post-infection clinico-immunologic syndrome: fundamentals of etiopathogenesis and immunogenodiagnostic strategy]. AB - Professor Yu.M. Lopukhin, an ardent advocate of translation of scientific achievements into clinical medicine, is a founder of new scientific disciplines including clinical immunology that evolved from the fusion of new knowledge, original clinical thought, high-tech diagnostic and therapeutic modalities. Highly promising investigations into molecular pathogenetic mechanisms of various diseases, their diagnosis, and prevention make up a most important aspect of his scientific work. The progress of chronic recurring infectious diseases (CRID) is closely related to the associated immune disorders, e.g. post-infection clinico immunologic syndrome (PIKIS) that reflects the character and severity of disturbances of immune homeostasis. This syndrome associated with CRID is as a rule provoked by infectious agents of different nature, immunopathologic processes inherent in individual patients, specific clinical course ofCRID or inadequate application of antimicrobial medicines. Three forms of PIKIS are described in this paper: (1) post-infection secondary immunodeficiency syndrome:, (2) post-infection autoimmune syndrome, (3) combination of the two. PMID- 20000104 TI - [Clinical efficacy of the biospecific hemosorbent "Liposorb"]. AB - The aim of the present work was to evaluate clinical efficacy of the biospecific hemosorbent "Liposorb". The main component of "Liposorb" is polyacrylamide gel containing an immobilized affine ligand (antibiotic polymyxin E-colistyne). 40 patients with abdominal sepsis and peritonitis of different genesis underwent a total of 52 seances of vein-venous extracorporeal hemoperfusion of hemosorbent "Liposorb". Endotoxin level (lipopolysaccharide - LPS) was measured by the turbodimetric method. Blood perfusion through a "Liposorb" column at 50-70 ml/min during 90 minutes permitted to reach stabilization of parameters of systemic hemodynamics. All the patients showed positive dynamics of general well-being, blood gas composition, clinical and biochemical blood analyses. The endotoxin (LPS) level of Gramm-negative flora significantly decreased. It is concluded that hemosorbtion using biospecific polymyxin-containing hemosorbent "Liposorb" effectively removes Gramm-negative endotoxin and leads to stabilization of hemodynamic in patients with Gramm-negative abdominal sepsis. PMID- 20000106 TI - [Iu. M. Lopukhin and the development of bioethics in Russia]. AB - Professor Yu.M. Lopukhin has made the most important contribution to the development of bioethics in this country based on his extensive experience in research, education and organization pertaining to different biomedical disciplines and high-tech medicine. The main works of Yu.M. Lopukhin on major bioethical problems of special importance for present-day Russia are analysed including ethical expertise of biomedical research and their legal aspects including tissue and organ transplantation, the use of embrionic stem cells, etc. The importance of international collaboration in the field of bioethics is emphasized. PMID- 20000105 TI - [Amino acid chloramines and chlorimines as antiplatelet agents: reactive properties and mechanism of action]. AB - Oxidative modifications of thiols, disulfide, and thioester atomic groups in proteins, peptides, and amino acids induced by chloramines or chloramine derivatives of amino acids and other reactive oxidants are considered. In the case of disulfide and thiol groups, production of sulfur-reactive groups may take place, such as disulphide S-oxides and sulphenic groups. Various chloramines and chloramines differently modify sulfur-containing groups. For example, N,N dichlorotaurine rapidly modifies the thiolgroup in reduced glutathione and N chloroglycine readily oxidizes the thioester group in methionine. Amino acid chloramines inhibit platelet aggregation by modifying S-containing centres. Autodecay of amino acid chloramines does not affect aggregation as follows from the absence of positive correlation between chloramines decay rate and antiplatelet activity. N,N-dichlorotaurine and its chlorimine derivatives are characterized by high stability and have good prospects as potential antiaggregants. PMID- 20000107 TI - [Determination of medium-weight molecules in the plasma and urine of neonatal infants with ischemic nephropathy]. AB - To ascertain the level of medium-weight molecules in the plasma and urine in critically ill neonatal infants with varying ischemic nephropathy, the authors determined the content of medium-weight molecules by N. I. Gabrielyan's procedure modified by M. Ya. Malakhova. Within the first 24 hours of life, the newborn infants with ischemic nephropathy had high plasma levels of medium-weight proteins. The high plasma levels of medium-weight molecules were attended by their urinary elimination in first- and second-grade ischemic nephropathy and these were not compensated for by their elimination in third-grade nephropathy, which was linked to decreased renal excretory function. The determination of medium-weight molecules in the plasma and urine permits identification of acute renal failure in the newborn. PMID- 20000108 TI - [Various acute phase reactants in different types of proliferative diseases of the uterine appendages]. AB - The authors examined serum in patients with ovarian cancer (OC; a disseminated process), ovarian cystadenoma (OCA), or external endometriosis (EM) before treatment and in apparently healthy females (a control) for the content of some acute-phase proteins and cytokines to clarify the specific features of changes in their concentrations in relation to the type of the proliferative process. It was shown that in OC, there were significant reductions in the levels of alpha2 macroglobulin (MG), plasmin (PL), alpha1-antitrypsin (AT) and statistically significantly increases in the content of lactoferrin (LF), interleukin (IL)-6, IL-8, Ig, and the regulatory transport complex of P--M. In M, the concentrations of AT were lower and those of IL-6, IL-8, and PL-MG were higher (to a lesser degree than those in OC). In OCA, the levels of MG and IgA were increased; those of IL-8 and PL-MG were decreased. The concentrations of interferon and IgM were unchanged in all groups. The findings suggest that difefrent proliferative processes initially provoke a number of changes of varying magnitude and even directions in the serum levels of inflammation reactants, which should be borne in mind when conducting clinical tests in the intraoperative and, probably, postoperative periods. PMID- 20000109 TI - [The specific features of the blood gas transport system in patients with postinfarction cardiosclerosis]. AB - The specific features of blood gas transport system functioning were analyzed in patients with cardiovascular diseases. In patients with postinfarction cardiosclerosis (PICS), the quantitative mechanism for hypoxia adaptation tended to decrease, which may be considered to be a compensatory-adaptive reaction aimed at eliminating the sludge phenomenon and improving the rheological characteristics of blood. Acute myocardial reinfarction developed in patents with PICS is characterized by the lower functional activity of red blood cells, and developing hypoxia is an important link of activation of apoptotic cell death. The degree of hypoxia may be believed to correlate with the sizes of a myocardial necrosis focus. PMID- 20000110 TI - [Diagnostic efficiency of laboratory tests determining the functional status of the endothelium in patients with chronic diffuse liver diseases]. AB - The patients with chronic diffuse liver diseases were found to have signs of endothelial damage, which manifests itself as increases in the count of desquamated endotheliocytes, the level of vascular endothelial growth factor, and the concentration of von Willebrand factor in plasma and to have signs of endothelial dysfunction as reduced nitric oxide levels and elevated endothelin-1 concentrations in plasma. The magnitude of changes in the indicators of endothelial dysfunction and in the markers of endothelial damage depends on the severity of hepatic damage. PMID- 20000111 TI - [A simple and highly sensitive high-performance liquid chromatography assay of salivary putrescine and cadaverine]. AB - A simple and highly sensitive high-performance liquid chromatography assay is proposed to test salivary diamines (putrescine and cadaverine). Derivation was carried out with the orthophthalic aldehyde 2-mercaptoethanol. A rapid purification procedure for derivatives on the cartridges packed with 10 mg of hypercrosslinked polystyrene (Purosep-200) was first developed. Separation was made on a Chromolith (Merck), 100 x 4.6 mm in size, with monolithic reverse-phase silica gel (RP-18e) in the isocratic mode with ultraviolet (UV) detection at 230 nm. The eluent contained 55% acetonitrile and 45% 0.01 M phosphate buffer pH 6.8, added by 1% of isopropanol; flow rate was 1400 pl/min; pressure was 28 bars. Complete separation of diamine derivatives lasted at least 5 min. The sensitivity of the assay with UV detection (230 nm) was about 0.1 ngfor diamines and about 0.5 ng for the internal standard (IS) at a signal/noise ratio of 3.0, which enabled diamines to be determined in I pl (0.001 ml) of saliva. The simplicity, reproducibility, and high sensitivity of the assay along with the feasibility of its application on standard chromatographic equipment (an isocratic pump and an UV detector) make it suitable for routine clinical application. PMID- 20000112 TI - [Statistical control of the quality of clinical laboratory analyses (a lecture)]. AB - No measurement can be absolutely precise, but, by using the methods of the probability theory and by knowing the nature of errors, one can estimate a probable site. The paper considers the specific features of measurements during clinical laboratory analyses, as well as those of statistical methods for assessing their validity. The first part contains general provisions; the second one outlines numerical methods for assessing the quality of performed work. The procedures using both the data of analyses carried out in patients and those of special studies of a control biological material of known and unknown composition. Most numerical methods suggest that a laboratory has at its disposal a computer connected to an analyzer, but a few, quietly reliable assessments can be also manually made. The paper gives only final conclusions and recommendations and omits rather intricate mathematical proofs, which are for information available at the web site clilab.ru in the section "Introduction to statistical quality control". PMID- 20000113 TI - [Effect of infectious factors on the activation of proinflammatory cytokines in chronic heart failure]. AB - The results of recent studies favor the immunological theory of the pathogenesis of chronic heart failure (CHF). In this connection, of particular interest is a search for possible causes of cytokine hyperactivation in patients with CHF of ischemic etiology, one of which may be viral infection. The aim of the study was to evaluate the impact of herpesvirus simplex (HVS), cytomegalovirus (CMV), Chlamydia pneumoniae on the activation of cytokines in patients with CHF of ischemic etiology. Ninety patients with CHF of ischemic etiology were examined. After comprehensive study, the patients were divided into 2 groups: 1) Functional Class (FC) I-II CHF and 2) FC III-IV CHF Increases in the levels of proinflammatory cytokines (tumor necrosis factor-alpha, interferon-gamma and in the content of IgG and IgM to CMV and IgM to HVS were ascertained in patients with CHF of ischemic etiology. There were statistically significant correlations of the level of proinflammatory cytokines, CMV, and HVS with the FC of CHF and the echocardiographic indicators reflecting left ventricular systolic function. PMID- 20000114 TI - [Evaluation of the immunoregulatory activity of neutrophilokine fractions in a macrophage model]. AB - A procedure was proposed to evaluate the immunoregulatory activity of neutrophilokine fractions on a model of macrophages. It was established that all the fractions studied did not affect the absorptive capacity of these cells in both primary and secondary immune responses. At the same time, the majority of neutrophilokine fractions modulated the killer activity of macrophages: they potentiated or inhibited it. The proposed procedure for evaluating the regulatory effect of individual neutrophilokine fractions on a model of studying the killer activity makes it possible not only to characterize their activity, but also to identify helper and suppressor fractions, which discloses approaches to correcting an immune response by means of these fractions. PMID- 20000115 TI - [Basic methods for extraction and molecular analysis of micromycetic DNA]. AB - The methods for extraction of nucleic acids from the fungi and clinical materials containing various microorganisms have currently modified and make it possible to extract DNA with its highest yield and to minimize its destruction. In particular, this concerns mycelial fungi that have a rigid cell wall. Addition of glass microbeads to the lysed sample, followed by mixture stirring, gives rise to undamaged DNA from fungi and their spores. Extracted DNA is suitable for the identification of micromycetes by qualitative or quantitative polymerase chain reaction (PCR), real-time PCR-based hybridization, and LightCycler PCR and for the polymorphism analysis of the spacer regions of ribosomal genes by the sequencing technique. A combination of quantitative PCR and sequencing permits co determination of the number of gene target copies in 9 different micromycetes in the clinical material being examined, which is of great importance for the diagnosis and treatment of patients with higher immunosuppression and, of those after transplantation of organs and tissues in particular. PMID- 20000116 TI - [Comparative study of the biological properties of various nutrient media used for the isolation of Legionella pneumophila]. AB - Three diagnostic selective media used for the isolation of Legionella pneumophila were compared. These included BCYEAalpha (Oxoid, a reference medium), BCYEAalpha (Hi Media), and elective legionellosis medium (ELM) developed at the Rostov-on Don Research Institute for Plague Control. The virulent L. pneumophila strain Philadelphia-1 (LD50 was 10(5) CFU for guinea-pigs) was used a test culture. The susceptibility of the media was determined, by culturing 10(-6) and 10(-7) dilutions of the suspension of the macerated Legionella-infected guinea-pig spleen, as well as the suspension of a culture of this strain (100 CFU) and 6 L. pneumophila cultures freshly isolated from water. The BCYEAalpha (Oxoid) and ELM media demonstrated the similar growth characteristics (chi2 < 0.7; p = 0.05) while the BCYEAalpha (Hi Media) medium showed a low susceptibility. The BCYEAalpha (Oxoid) and ELM media were first found to be successful in detecting Legionella in viable, but nonculturable state, induced by the following factors: 1) starvation in distilled water; 2) exposure to hydroquinone (oxidative shock, and 3) elevated temperature (56 degrees C). The BCYEAalpha (Oxoid) and EML media did not differ either in their ability to suppress extraneous microflora and to maintain stable initial pH under the conditions of incubation of culture plates, as well as in their Na+ concentration (15-19 mmol/l). However, the BCYEAalpha (Oxoid) medium exceeded the ELM one in the growth rate and diameter of Legionella colonies. Two L. pneumophila cultures were isolated in the BCYEAalpha (Oxoid) and EML media used in the field experiment studying 15 water samples from different hot water supply systems. Thus, the conclusion can be drawn that the EML medium is comparable with the reference BCYEAalpha (Oxoid) medium in its susceptibility and ability to detect Legionella in both vegetative and viable, but nonculturable states and is suitable for practical application. PMID- 20000117 TI - [Osmometry]. AB - The osmotic concentration, or osmolality, of biological fluids is measured by means of osmometers-cryoscopes, whose operating principle is based on cryoscopy, namely on the determination of the osmolality of solutions from the change in their freezing point. The osmometers-Cryoscopes can determine the effective osmolality that is created by the inorganic molecules of a solution. Stable osmolality in the body fluids is provided by an osmotic pressure Regulatory system. The osmometers have presently extensive applications in different areas of medicine, pharmacology, and food-processing industry. PMID- 20000118 TI - [Glutathione system. II. Other enzymes, thiol-disulphide metabolism, inflammation and immunity, functions]. AB - The great significance of glutathione as a redox regulator and the reducing carrier has been established. There is a clear necessity for subdivision of an independent mitochondrial glutathione subsystem. The data on a specificity of glutathione metabolism in different organs are accumulated. The significance of glutathione system for inflammation and immunity has been proved. The investigations of glutathione system for elucidation of pathogenesis of diseases and its diagnostics are used in medicine. PMID- 20000120 TI - [Structure and physiological significance of angiotensin converting enzyme domains]. AB - Somatic angiotensin converting enzyme (ACE) consists of two homologous catalytic domains (N- and C-domain), each of which bears an active site exhibiting different biochemical properties. The ACE isoforms consisted of one domain were also detected in mammals. Substantial progress in ACE domain research was achieved during the last years, when crystal their structures were determined. The crystal structures of domains in complex with diverse potent ACE inhibitors provided new insights into structure-based differences of the domain active sites. Physiological functions of ACE are not limited by regulation of the cardiovascular system. Recent evidence suggests that the ACE domains may be also involved into control distinct physiological functions. The C-terminal catalytic domain, playing important role in regulation of blood pressure, catalyzes angiotensin I cleavage in vivo. The N-domain contributes to processing of other bioactive peptides for which it exhibits high affinity. Domain-selective inhibitors able to block selectively either the N- or C-domain of ACE have been developed. PMID- 20000119 TI - [Comatose states: etiopathogenesis, experimental studies, treatment of hepatic coma]. AB - The review presents the modern concepts on biochemical mechanisms of processes, that result in comatose states (CS), with emphasis on the search of new therapeutic approaches. CS of various origin causes severe suppression of brain cells functioning and stable unconsciousness. Numerous reasons of various CS are classified into two main groups: primary brain damages (ischemia, tumor, trauma) and secondary damages originating from system injuries in the body (endocrine, toxic e. c.). The most often primary CS is the hypoxic-ischemic one, as result of corresponding encephalopathy. Its mechanism is the brain cells "energy crisis"- because of decreased blood supply or its deficiency by energy substrates or/and by oxygen. Among secondary CS the substantial place takes hepatic coma as a consequence of hepatic encephalopathy in severe liver diseases--cirrhosis, acute liver failure, sharp intoxication. Its main reason is associated with exess of ammonia entering the brain tissue (it accumulates in blood because of lack of its removing by damaged hepatocytes). Ammonia reacts with glutamate in brain astrocytes and the product of this reaction, glutamine, induced osmotic imbalance, that results in change of form and functions of these important brain cells. It induces, in turn, neurons functions damages, changes in neurotransmission and cerebral blood flow and all these may give rise CS. The most of CS studies are carried out in human. Experimental models ofhepatic CS are reproduced mainly in rats, the most often by surgery methods. Other models included administration of thioacetamide or D-galactosamine, sometimes in combination with lipopolysaccharide. In earlier studies ammonia administration together with liver damages by ligation or by CCl4 was used. The main principles of hepatic coma treatment include the care of encephalopathy, detoxification, and liver treatment. Elaboration of new nanodrugs with increased penetration into tissues and cells, in particular, on the base of phospholipid nanoparticles, may increase substantially the therapeuti efficiency. One of such drug is thought to be a new hepatoprotective preparation phosphogliv--nanoparticles of soy phosphatidylcholine with glycyrrhizic acid. It is supposed, that the further development of phospholipid nanoforms, with minimal particle sizes, may reveal the more action in CS treatment. PMID- 20000121 TI - [Biomarkers in the modern medical and biologic practice]. AB - The lecture deals with biomarkers, a newly introduced direction in biomedical research. The following terms, definitions, and characteristics were proposed to describe biomarkers: diagnostic tools, clinical endpoints and surrogate endpoints. Examples of predictors for cardiovascular diseases, acute-phase of inflammation and malignant tumors are represented. The special attention is given to the characteristic of the widespread biomarkers: homocysteine, transthyretin and C-reactive protein. Modern methods of identification, qualitative and quantitative determination of biomarkers are considered: mass spectrometric methods, immunoassay. The relationships between the latest achievements in the field of genomics, proteomics, bioinformatics and biomarker's investigations are established. The perspectives of biomarkers are discussed. PMID- 20000122 TI - [Expression of gelatinases A and B and their endogenous regulators in immortal and transformed fibroblasts]. AB - Matrix metalloproteinases (MMP) play a critical role in tumor invasion and metastasis. The aim of this study was to elucidate peculiarity of expression of gelatinases A and B (MMP-2 and MMP-9), membrane type MMP (MT1-MMP) and tissue inhibitor of MMP (TIMP-2) in immortal (IF) and transformed fibroblasts (TF).The study was carried out using embryo rat fibroblasts, sequentially immortalized with the polyomavirus LT gene and transformed with the E7 gene of human papilloma virus (HPV-16). Papilloma virus type 16 and 18 are etiological factors of cervical cancer. The primary fibroblast (PF) culture of Fisher rats was used as control. Analysis of TF and IF involved: determination of MMP-2 and MMP-9 activity by hydrolysis of specific substrate--radioactive collagen type IV; obtaining of MMP spectra by zimographic assay and estimation of the mRNA expression (by RT-PCR) of MMP-2, MMP-9, MT1- MMP and TIMP-2. It was found: 1) collagenolytic activity of MMP was increased only in TF and was dependent on the degree of cell tumorogenity; 2) the study of MMP spectra was shown that MMP-9 was found in TF only but MMP-2 was found in all investigated clones; 3) The mRNA expression of MMP-9, MT1-MMP and TIMP-2 was increased in all TF while the MMP-2 expression was increased in TF only after TF cell selection on rats; 4) The collagenolytic activity as well as the mRNA expression of MMP-2 and MMP-9 themselves and of MMP-2 endogenous regulators (MT1-MMP and TIMP-2) did not change in immortalized fibroblasts compared to PF. The data obtained indicate changes in the enzyme/inhibitor/activator ratio and also suggest of a significant increase in the TF destructive potential. MMP-9 is supposed to be a marker of fibroblasts transformed by E7 HPV-16 gene in cell culture. PMID- 20000123 TI - [The effect of myosin ATPase inhibition on metabolic and functional recovery of isolated rat heart after global ischemia]. AB - The effect of myosin ATPase inhibitor, 2,3-butanedione monoxime (BDM; used in the range of concentrations 1.25-10.0 mM), on recovery of functions of isolated rat heart subjected to normothermic (37 degrees C) total ischemia for 35 min has been investigated. BDM perfusion was performed at a flow rate of 4 ml/min during 5 min before ischemia (BDM-I) or before 25-min reperfusion (BDM-R). Control hearts were perfused with Krebs solution at the same flow rate. The highest functional recovery of heart and coronary vessels was observed during infusion of 2.5 mM BDM before ischemia. At the end of reperfusion ATP and phosphocreatine (PCr) content in hearts of this group was significantly higher whereas the level of lactate was two times lower than in control; total creatine content (sigmaCr) did not differ from the initial level. Similar but less pronounced changes in the improvement of aerobic metabolism and maintenance of sigmaCr after reperfusion were also observed in the case of infusion of 2.5 mM BDM before reperfusion. They were consistent with reduced recovery of functions of heart and coronary flow compared with these parameters observed in the BDM-I group. 2.5 mM BDM caused almost 2 fold decrease in release of cardiac lactate dehydrogenase into myocardial perfusate in the BDM-I and BDM-R groups (compared with control); this suggests lower damage of cell membranes. These results suggest that improvement of energy supply of postischemic cardiomyocytes may be a key factor determining cardioprotector effectiveness of short-term administration of BDM before ischemia. PMID- 20000124 TI - [Biosensor analysis of interaction of potential dimerization inhibitors with HIV 1 protease]. AB - Currently inhibitors of protein-protein interactions are considered as perspective prototypes of new generation of drugs. The most attractive targets for such inhibitors are the oligomeric enzymes which active sites are formed by amino acid residues from different subunits. The classic example of such enzyme is HIV protease (HIVp), active only in the homodimeric form. We have developed a new approach for experimental screening of HIVp dimerization inhibitors. It is based on the original biosensoric test-system for differential analysis of interaction of tested substances with HIVp dimers and monomers. Using this test system we have analyzed the most perspective candidate substances predicted by method of virtual screening, and some derivatives of glycyrrhizin, triterpenic and steroid glycosides. As a result we found one compound, which mainly interacts with HIVp monomers and inhibits in vitro activity of this enzyme with IC50 of about 10(-6) M. PMID- 20000125 TI - [Biodegradable polymer microparticles with entraped herbal extracts: preparation with supercritical carbon dioxide and use for tissue repair]. AB - Biodegradable microparticles based on poly-D,L-lactide with entrapped mixture of herbal water-soluble extracts of Plantago major and Calendula officinalis were prepared. For preparation of these microparticles the previously developed method based on the usage of supercritical carbon dioxide (SC-CO2) was proposed. Microparticles were obtained by two techniques: 1) by preparing porous polymer monolith containing entrapped mixture of herbal extracts, which was then reduced to fine microparticles (ca. 0.1 mm) by dry ice grinding (called here as "monolithisation technique") and 2) by spraying of this polymer/extracts mixture through a jet (spray technique). In vitro release kinetic profile of herbal extract mixture was found to depend on the microparticle preparation technique, on the microparticle structure as well as on the initial ratio polymer/extracts (w/w). The microparticles were used for gastric ulcer treatment in a rat model. The extracts released from microparticles were found to accelerate tissue repair. PMID- 20000127 TI - [Correction of glutamine metabolism impairments in the operated liver with chronic hepatitis by hyperbaric oxygen]. AB - Application of hyperbaric oxygenation (HBO, 3 ata, 1 session for 50 min per day) during the first three days after liver resection (LR, 15-20% from the organ mass) in animals with chronic toxic hepatitis (CCl4, 50%, 0,1 ml/per 100 g of body mass, subcutaneously, once in 2 days, 65 days) eliminates a deficit of glutamine and glutamate in an operated liver and prevents accumulation of the endogenic toxin, ammonia, caused by combined effects of CCl4 and LR. Thus hyperbaric oxygen modulates the effect of the LR on the activity of key enzymes of the glutamine metabolism in liver: glutamine synthetases (GS) and phosphate dependent glutaminases (PDG). HBO enhanced and prolonged the LR effect of the GS activity and restricted analogous changes in PDG during an early (3 day) postoperative period and promoted a delayed transient stimulation in the late (7 day) postoperative period. In contrast to non-oxygenated animals with LR this was not accompanied by accumulation of ammonia and the decrease in glutamine concentration in the liver. PMID- 20000126 TI - [Comparative characterization of catalytic properties of mitochondrial and cytoplasmic forms of NAD-dependent malate dehydrogenase from the rat liver at norm and in toxic hepatitis]. AB - The purification and comparative characterization of some catalytic properties of liver mitochondrial and cytosolic NAD-dependent malate dehydrogenase (NAD-MDH; EC 1.1.1.37) from normal rats and rats with experimental toxic hepatitis (ETH) have been carried out. It has been found that there are some differences in catalytic and regulatory properties of liver NAD-MDH from control animals and rats with ETH. It has been shown that Fe2+ and Cu2+ ions inhibit the enzyme, and the inhibition degree is different at norm and under toxic hepatitis. Ca2+ ions insignificantly activate cytosolic NAD-MDH under pathology and do not influence the mitochondrial isoform. PMID- 20000128 TI - [The relatioship between decomposition of amino acid chloramines and their structures]. AB - Rate constants of the decomposition of monoamine alpha-amino acid chloramine derivatives were determined by a spectrophotometeric method. Several amino acid chloramines with elevated stability have been found. These included n chloroglycine, n-chlorovaline, n-chlorothreonine, and n-chloroisoleucine. Their molecular structures are characterized by some characteristic feature at the beta position. In the case of glycine chloramine, carbon atom at this position is absent, and the chloramine derivatives of three other amino acids possess branched chains. Partial atomic charges of the electrostatic potential (Wang Ford) for chloramines of alpha-amino acids were computed using the semiempirical quantum-mechanical method AM1. The chloramines with elevated stability have high positive sums of charges of three carbon atoms that are atoms at alpha- and beta positions and a carboxyl group atom. High partial charge also was obtained for one carbon atom at the beta-position. These computational values may be employed for prediction of the stability of designed amino acid chloramines. One of the important predictions is that the highest atomic charges and stabilities are characteristics of the amino acid chloramines, in which all hydrogen atoms at the beta-position are replaced by carbon-hydrogen chains or hydroxyl groups. PMID- 20000129 TI - [Anticonvulsants as bioantioxidants under stress conditions]. AB - The action of heterocyclic amides series (ethosuximide, phenytoin, primidone) on lipid peroxidation and membrane bound monoamine oxidases A and B under stress condition has been studied. The intraperitoneal injection of the drugs resulted in enhancement of SOD, decrease of brain malondialdehyde content and mitochondrial activity of monoamine oxidases A and B. PMID- 20000130 TI - [Phospholipids of blood and lymphocytes membranes in virus hepatitis B and toxic hepatitis]. AB - Phospholipid composition of blood plasma and lymphocyte membranes was investigated in patients with HBV infections and toxic hepatitis associated with the use of surrogate alcohol products. The revealed changes in phospholipids spectrum of blood plasma and lymphocyte membranes reflect opposite changes in biochemical processes influenced by a virus infection (hepatitis B) and toxic hepatitis. PMID- 20000131 TI - Recommended practices for femtosecond laser in refractive surgery. PMID- 20000132 TI - Quick as a wink. Tips for administering ophthalmic topical medications. PMID- 20000133 TI - Managing the care of the obese ophthalmic patient. PMID- 20000134 TI - Accepting diabetes and denying blindness ... as seen through a juvenile's eyes. PMID- 20000135 TI - Helpful hints for clinical research coordinators. PMID- 20000136 TI - Jurdean Reed, RN--ophthalmic nurse extraordinaire. PMID- 20000137 TI - Leading the way to medical office success during difficult times. PMID- 20000138 TI - Most eye injuries in school-age children are sports-related. Prevent Blindness America urges public to make eye safety while playing sports a priority. PMID- 20000139 TI - Nearly half of all eye injuries occur in or around the home. Data shows number of eye injuries to children on the rise. PMID- 20000140 TI - A guided tour of PubMed for ophthalmic nurses. PMID- 20000141 TI - Vision loss and visual hallucinations: the Charles Bonnet syndrome. PMID- 20000142 TI - Respecting our elders. PMID- 20000143 TI - Will nurses who do not have a degree be left behind on the career ladder? PMID- 20000144 TI - Is the expenditure on temporary staff justified? PMID- 20000145 TI - Easing the pain of loss. AB - Streamlining services for women who lost a baby in the second or third trimester was the goal of maternity care assistant Lyndy Swain. PMID- 20000146 TI - Ask the difficult questions. AB - We all make judgements about people based on pre-conceived ideas. Recognising individuality is key to avoiding discrimination. PMID- 20000147 TI - A reason to hope. AB - The recent announcement that vaccination against HIV has been partially successful in Thailand gives scientists important clues for further developments. PMID- 20000148 TI - Screening hits the streets. AB - Innovative practice using peer educators is taking HIV testing to sex workers in Ghana. PMID- 20000149 TI - Culture of hate. PMID- 20000150 TI - Be receptive to the signals that mean someone wants to talk. PMID- 20000151 TI - Provision of end of life care in the community. AB - Many people who are terminally ill would prefer to die at home. Integrated professional working is necessary to enable patients to remain in their homes for end of life care. This article describes how an integrated district nursing team in Lincolnshire provides end of life care to patients in the community. The team uses the Gold Standards Framework and the Liverpool Care Pathway to provide optimum care to patients and their families and carers at the end of life. PMID- 20000152 TI - Treatment of older patients with aortic valve stenosis. AB - This article aims to increase knowledge and understanding of aortic valve stenosis to support the treatment options available for older patients in the acute setting. The article discusses a relatively new procedure, transcatheter aortic valve implantation, which is appropriate for selected patients with aortic stenosis, and outlines the aftercare involved. PMID- 20000153 TI - An introduction to electrocardiogram interpretation: Part 1. AB - This article describes how to interpret the single-lead electrocardiogram (ECG). The author also describes cardiac electrophysiology and how this is represented by ECG graphs. Part 2 of this article, to be published next week, will describe 12-lead ECGs and acute coronary syndromes. PMID- 20000154 TI - Care after a death. PMID- 20000155 TI - Working through your emotions. PMID- 20000156 TI - How to read a job advertisement. PMID- 20000158 TI - Gestational diabetes. PMID- 20000159 TI - Breast tomosynthesis. PMID- 20000160 TI - Vaginal pH as an OTC screening tool for vaginitis. PMID- 20000161 TI - Treating otitis media in retail clinic settings. PMID- 20000162 TI - Watchful waiting. A viable approach to acute otitis media. PMID- 20000163 TI - Double dose. Two flu vaccines on tap this season. PMID- 20000164 TI - Choosing a healthcare plan. PMID- 20000165 TI - Prescription for respect. Drug companies should value NPs. PMID- 20000167 TI - Patient care paradox. Why is retail health not serving the underserved? Interview by Jennifer Ford. PMID- 20000168 TI - More from the trenches. Answers to common billing questions. PMID- 20000170 TI - Delirium in older adults. PMID- 20000171 TI - Bioidentical hormones. PMID- 20000172 TI - Pneumonia in older adults. PMID- 20000173 TI - Treating pneumonia in the retail setting. PMID- 20000175 TI - Much more than a nuisance. Health consequences of sleep disorders. PMID- 20000174 TI - Acute HIV infection in primary care. PMID- 20000176 TI - Just say no. Rethinking flattery, food, freebies. PMID- 20000177 TI - The solution is in place. PMID- 20000179 TI - Detecting hearing loss in infants. PMID- 20000180 TI - Novel cardiac markers. PMID- 20000181 TI - Fluid transport gone wrong. An introduction to lymphedema. PMID- 20000182 TI - Winning the battle of the bulge. Practical approaches to weight loss. PMID- 20000183 TI - Initial therapy for type 2 diabetes. Examination of a combination approach. PMID- 20000185 TI - Varicose veins. Today's treatment tools. PMID- 20000184 TI - Health needs of collegiate men. PMID- 20000186 TI - Hypertension in children and adolescents. PMID- 20000187 TI - What's your mantra? PMID- 20000188 TI - Saddle up. The real value of a nurse practitioner. PMID- 20000189 TI - Dental amalgam. PMID- 20000190 TI - Dental caries experience of children in Northland/Te Tai Tokerau. AB - OBJECTIVE: To describe the prevalence, severity and intra-oral distribution of dental caries among children from four Northland communities. DESIGN: Cross sectional epidemiological survey of a Census sample. METHODS: Dental examinations were systematically conducted on all children aged 5 or 6 and all children 12 or 13 (and for whom parental consent was obtained) in Kaitaia, Kaikohe, Kawakawa/Moerewa and Dargaville. Bitewing radiography was used to aid caries diagnosis in the older age group. Data are reported for the deciduous dentition in the 5-6-year-olds, and the permanent dentition in the 12-13-year-olds. RESULTS: There were 369 5-6-year-olds and 171 12-13-year-olds, approximately two thirds of whom were Maori. Almost 88% of 5-6-year-olds had experienced dental caries, and this was lower among the Pakeha/Other group, those attending higher decile schools, and those from Dargaville. Overall caries experience (represented by a mean dmft score of 5.6; sd 3.9) was higher among Maori children, those attending lower-decile schools, and those from Kaikohe, Kawakawa/ Moerewa or Kaitaia, as was the number of untreated decayed surfaces. Almost one in four children had lost at least one tooth due to caries, and this was higher among Maori children and those from schools in more deprived areas, and lower among children from Dargaville. Caries experience was greatest among the second molars in either arch, and lowest among the mandibular incisors. Among the 12-13-year olds, 85% had experienced dental caries (mean DMFT = 3.7; sd 3.1). The number of untreated surfaces with decay (more than 3, on average) was considerably lower among children from Dargaville. Caries experience was greatest among the first molars in either arch (followed by the second premolars), and absent among the mandibular incisors. CONCLUSIONS: The prevalence and severity of dental caries in Northland is very high in comparison to the rest of New Zealand. The problem is particularly acute among northern communities and Maori. Dealing with the problem will require a combination of population health and personal health interventions. PMID- 20000191 TI - The prevalence of Molar-Incisor Hypomineralisation (MIH) in Wainuiomata children. AB - OBJECTIVES: The aim of this study was to determine the prevalence of Molar Incisor Hypomineralisation (MIH) in Wainuiomata children and describe differences in prevalence among Maori, Pacific Island and New Zealand European ethnic groups. DESIGN: Cross-sectional survey of developmental defects of enamel in a random sample of children attending primary school in Wainuiomata, Wellington. METHOD: Study information and consent forms were sent to 850 7-to-10-year-old schoolchildren. Using the modified Developmental Defects of Enamel index, a single paediatric dentist examined students in the classroom. Dental caries experience was recorded as decayed, missing or filled primary and permanent teeth. RESULTS: Examinations were conducted on 522 children (participation rate 61.4%). The mean age of the children was 8.2 years (range 7 to 10 years). MIH prevalence was 14.9%. The prevalence ofhypomineralisation ofany tooth was 15.3%, and that for hypoplasia was 4.0%. There was no statistically significant ethnic difference in MIH prevalence. The mean DMFT was 0.16 (SD, 0.54) in those without a developmental defect, 0.54 (SD, 1.12) in those with hypomineralisation and 1.85 (SD, 1.85) in those with hypoplasia (p < 0.01). CONCLUSIONS: Approximately one in seven Wainuiomata children have MIH. Ethnicity is not a modifying factor in the occurrence of developmental defects of enamel. The presence of hypomineralisation and/or hypoplasia was associated with significantly greater caries experience in the permanent dentition. PMID- 20000192 TI - An interesting case of recurrent temporomandibular joint dislocation. AB - Acute temporomandibular joint dislocation is an infrequent presentation to the emergency department or to dental practitioners. It may be a rare complication of dental treatment. Recurrent temporomandibular joint dislocation occurs in individuals with certain anatomical abnormalities or systemic conditions. This paper reports an unusual presentation of recurrent temporomandibular joint dislocation and outlines some of the surgical techniques used in managing this condition. PMID- 20000193 TI - Tooth shade selection using a colorimetric instrument compared with that using a conventional shade guide. AB - OBJECTIVES: The aim of this study was to compare the colour matching ability of instrumental colorimetry with that of a conventional visual colour matching technique. SETTING: The study was conducted in a dental clinic at the University of Otago, School of Dentistry, New Zealand. MATERIALS AND METHODS: Three dentists with normal colour vision used a Vita Classical shade guide to determine the shade of the maxillary right central incisor of 10 randomly chosen individuals. The tooth colour of the same teeth was then determined with the Shade Vision colorimeter. The examiners were not aware of one another's shade selection and colorimetric readings. The shades selected were assigned numeric Delta E values for data compilation and statistical analysis, and the degree of examiner reliability was determined using the intraclass correlation coefficient (ICC). RESULTS: Analysis of the data indicated good overall interexaminer reliability for the Vita Classical shade guide (ICC = 0.62); however, the data showed poor agreement between the two shade selection techniques (ICC = 0.27). CONCLUSION: This study found no significant difference in the accuracy of shade selection when using either a conventional visual assessment technique or a colorimetric instrument. There was, however, some discrepancy when the two shade selection techniques were compared for shade selection of the same tooth. The clinical relevance of this difference is unclear, suggesting that additional research is needed to determine the clinical advantage of using a colorimeter. PMID- 20000194 TI - Summary of guidance for the use of fluorides. PMID- 20000195 TI - MOST CITED: Number 7. Effect of fluoride and of soil trace elements on the morphology of the permanent molars in man. PMID- 20000196 TI - [Isolating, culturing and characterizing stem cells of human dental pulp origin]. AB - Evidence has been accumulating for the presence of stem cells in dental tissues. The authors' studies aimed to produce primary culture from human dental pulp. Furthermore, they wanted to identify clonogenic cells with progenitor properties in these cultures, and to characterize their proliferative capacity. The dental pulp was isolated from surgically removed wisdom teeth. The extracellular matrix was enzymatically degraded to obtain isolated cells for culturing. Identification of STRO-1 mesenchymal stem cell marker was achieved by immunocytochemistry. Osteogenic differentiation was detected by the application of Alizarin Red. The proliferative activity of the cell cultures in response to serum, EGF and BMP2 was estimated by MTT assay. The authors' most important finding is the successful establishment of stable primary cell culture from human dental pulp tissue. The cultures can be passaged multiple times and they contain clonogenic, STRO-1 immunopositive cells. Their mineralization capacity was shown by mineralized deposits as a result of induction by suitable medium. The presence of serum increased, while both EGF and BMP2 concentration-dependently decreased the cell proliferation in the cultures. The authors' model provides the foundation for studies of the proliferation and differentiation of dental pulp cells at molecular level, and opens a new direction towards the biological regeneration of dental tissues. PMID- 20000197 TI - [Clear cell adenocarcinoma of the tongue. Coincidence or distant metastasis?]. AB - Malignant tumors of the salivary glands are rare, they account for less than 0.5% of all malignancies and for 5% of cancers of maxillofacial regions. Clear cell adenocarcinoma represents 1% of malignant tumors of the salivary glands. The present case involved a 63-years-old Hungarian woman with a painless, non ulcerated, nodular mass on the right side of the middle part of the tongue. Her past medical history included right nephrectomy of renal cell carcinoma, seven years before the tongue tumor. The initial orofacial clinical examination suspected lipoma, neurinoma or distant metastasis of the renal cell carcinoma. The histomorphological analysis of the resected tongue tumor showed similar clear cell adenocarcinoma mimicking the phenotype of renal cell carcinoma but detailed immunohistochemical analysis proved the primary origin of tongue indicating second primary metachronous malignant tumor. PMID- 20000198 TI - [A rare case of fatal outcome due to extensive facial trauma caused by dogbite]. AB - Statistically 10% of head and neek trauma is caused by animal bite, more often inflicted by dogs. More than 50% of victims are infants and small children. Generally, the result of dogbite is superficial trauma but there are cases of deep soft tissue defects with amputation and multiple bone fractures. In case described in this article, an elderly woman suffered a dog attack, wich resulted in extensive face and head injuries. In spite of receiving proper treatment, the patient passed away. It is highly probable that underlying Syncumar therapy caused complications which contributed to the fatal outcome. Because of it's unusual nature we found this case to be of interest for publication. PMID- 20000199 TI - [In memoriam Vilmos Vajna: to commemorate his 155th birthday]. AB - Vilmos Vajna was born in Transylvania. His medical education was completed at Franz Joseph University of Arts and Sciences in Kolozsvar. He studied dental sciences in Vienna and Pest. After returning to Kolozsvar he obtained his habilitation ("venia legendi") and then lectured dentistry at this university. After finishing his academic carrier he moved to Budapest in 1895. While he practiced dentistry in his private office he made several technical innovations, constructed new instruments, and invented new technologies. He was an active member of the public life of the dental community. PMID- 20000200 TI - [Remnant of deciduous tooth]. PMID- 20000201 TI - National emergency declaration creates H1N1 options for EDs. AB - ED managers will have more latitude to deal with H1N1 surges thanks to President Obama's recent declaration of a national emergency. Here are some strategies that will help you keep things running smoothly. Determine your department's "tipping point," beyond which you must consider regulatory waivers to maintain efficient flow. Arrange for alternate treatment sites off-campus. Design a simplified screening process in the event of a medical screening exam (MSE) waiver. PMID- 20000202 TI - ED-centric approach earns hospital award. AB - Having an ED physician in charge of a hospital, or at least having the facility be sensitive to the specific challenges of the ED, can significantly improve performance. The success of this ED-sensitive approach is demonstrated by the awarding of the first Emergency Center of Excellence award to Aurora BayCare Medical Center in Green Bay, WI. The design of the ED space reflects the unique needs of ED patients and families. For example, a separate grieving area has been created. Other departments, such as radiology, are "trained" to give ED needs priority. Problems affecting the ED are discussed by all departments involved in those processes. PMID- 20000203 TI - At this center, 'staff' training means everyone. AB - As the staff of the chest pain center at Oregon Health & Science University Hospital moved toward reaccreditation, they paid careful attention to details. The ED valet received training in who to call when a chest pain patient arrives. The patient access service clerk was provided with drawings indicating where symptoms typically appear. Triage nurses received training in 12-lead EKGs and acute coronary syndrome. PMID- 20000204 TI - Low-protein latex gloves: a solution? PMID- 20000205 TI - The power in numbers. PMID- 20000206 TI - Testosterone replacement. PMID- 20000207 TI - Men want to look good, too. PMID- 20000208 TI - Getting paid for what you do. PMID- 20000209 TI - Bartholin cyst abscess. PMID- 20000210 TI - Patient information. Planning for pregnancy. PMID- 20000211 TI - Breast cancer and the family tree. An issue for all practice settings. PMID- 20000212 TI - The motivation to move. Prescribing exercise in primary care. PMID- 20000213 TI - Inside and out. A review of otitis in children. PMID- 20000214 TI - Rheumatoid arthritis. Understanding joint damage and inflammation. PMID- 20000215 TI - Skin is in. Laser therapy in primary care. PMID- 20000216 TI - How two NPs changed managed care. The birth of Evercare. Interview by Jill Rollet. PMID- 20000217 TI - Vitamin D. Research suggests broader health role. PMID- 20000219 TI - Ace the interview. Preparation is key. PMID- 20000218 TI - Stem cell therapy. Push politics aside to see human story. PMID- 20000220 TI - Suddenly independent. PMID- 20000227 TI - Pandemic: 2009 influenza A (H1N1). PMID- 20000228 TI - This issue: focus on influenza. PMID- 20000229 TI - A 2 1/2-year-old boy with leg pain. PMID- 20000230 TI - A 14-year-old girl with a patch of hair loss. Temporal triangular alopecia. PMID- 20000231 TI - Influenza vaccines: the key to disease prevention and control. PMID- 20000232 TI - The daunting practicalities of in-office pediatric influenza vaccination: 2009 2010. PMID- 20000233 TI - Influenza vaccination of healthcare personnel. PMID- 20000234 TI - Antiviral treatment and prophylaxis of influenza virus in children. PMID- 20000235 TI - Swine influenza viruses and their pandemic potential. PMID- 20000237 TI - Should we repair rotator cuff tears earlier? PMID- 20000236 TI - Radiologic case study. Spontaneous resolution of a spinoglenoid notch gang. PMID- 20000238 TI - Francis Xavier Dercum. PMID- 20000239 TI - Explosions and blast injuries II. PMID- 20000240 TI - A simple, reproducible method for centering the guide-pin in hip resurfacing arthroplasty. PMID- 20000241 TI - Osteonecrosis of the jaw associated with bisphosphonate therapy. PMID- 20000242 TI - Subtalar dislocation. PMID- 20000243 TI - Midshaft clavicle fractures: are surgical indications changing? PMID- 20000245 TI - Embedding quality and safety competencies in nursing education. PMID- 20000246 TI - Realist evaluation as a framework for the assessment of teaching about the improvement of care. AB - Health professions education researchers continually search for tools to measure, evaluate, and disseminate the findings from educational interventions. Clinical teaching, particularly teaching about the improvement of care and systems, is marked by complexity and is invariably influenced by the context into which the intervention is placed. The traditional research framework states that interventions should be adjudicated through a yes or no decision to determine effectiveness. In reality, educational interventions and the study of the interventions rarely succumb to such a simple yes or no question. The realist evaluation framework from Pawson and Tilley provides an explanatory model that links the context, mechanisms, and outcome patterns that are discovered during implementation of a project. This article describes the unique qualities of the realist evaluation, the basic components and steps in a realist evaluation, and an example that uses this technique to evaluate teaching about improvement of care in a clinical setting. PMID- 20000247 TI - Commentary on "realist evaluation as a framework for the assessment of teaching about the improvement of care". PMID- 20000249 TI - Student learning outcomes after integration of quality and safety education competencies into a senior-level critical care course. AB - Nursing education must respond to reports from the Institute of Medicine and others that health care education must focus more on safety and quality so graduates are empowered to positively impact patient safety. Effective teaching strategies must be developed and tested that result in positive student outcomes. The purpose of this pilot study was to test the effects of structured classroom and clinical content related to safety and quality of health care systems on a group of senior-level nursing students. A mixed-method study was conducted using repeated-measures analysis of variance quantitative data from the Student Perception of Safety and Quality Knowledge, Skills, and Attitudes Questionnaire and content analysis for the qualitative data collected from focus group discussions. Results suggest a combination of classroom and clinical learning activities have the strongest impact on student knowledge, skills, and attitudes related to safety and quality. PMID- 20000248 TI - Web-based hazard and near-miss reporting as part of a patient safety curriculum. AB - As part of a patient safety curriculum, we developed a Web-based hazard and near miss reporting system for postbaccalaureate nursing students to use during their clinical experiences in the first year of their combined BS-MS advanced practice nurse program. The 25-week clinical rotations included 2 days per week for 5 weeks each in community, medical-surgical, obstetrics, pediatrics, and psychiatric settings. During a 3-year period, 453 students made 21,276 reports. Of the 10,206 positive (yes) responses to a hazard or near miss, 6,005 hazards (59%) and 4,200 near misses (41%) were reported. The most common reports were related to infection, medication, environmental, fall, and equipment issues. Of the near misses, 1,996 (48%) had planned interceptions and 2,240 (52%) had unplanned interceptions. Types of hazards and near misses varied by rotation. Incorporating hazard and near-miss reporting into the patient safety curriculum was an innovative strategy to promote mindfulness among nursing students. PMID- 20000250 TI - Using quality and safety education for nurses to enhance competency outcome performance assessment: a synergistic approach that promotes patient safety and quality outcomes. AB - As nursing programs respond to vital initiatives such as Quality and Safety Education for Nurses (QSEN), nursing faculty will discover important shared values exist between competency-based curricular models and the latest call for stronger foci on safety and quality. This article describes how one university is using the QSEN competencies to enhance its competency outcome performance assessment (COPA)-based curriculum, thereby updating and strengthening its graduates' skills in quality improvement and safety. Faculty at the school found QSEN and COPA share the same concerns for promoting student competence and continuing competence in nursing practice to safeguard patient safety and quality care. Nursing faculty whose teaching is focused in a competency-based curriculum are well positioned to respond to the call to integrate QSEN competencies into their curricula. PMID- 20000251 TI - The new fundamentals in nursing: introducing beginning quality and safety education for nurses' competencies. AB - This article describes the redesign of the fundamentals of nursing course using an organizing framework and teaching strategies identified in the Quality and Safety Education for Nurses (QSEN) initiative. Six QSEN competencies (patient centered care, teamwork and collaboration, evidence-based practice, quality improvement, safety, and informatics) are essential for nursing practice. Beginning knowledge, skills, and attitudes (KSAs) associated with each competency were identified in a preliminary Delphi survey as important to incorporate early in prelicensure nursing curricula. Redesign requires a shift in focus from task training and psychomotor skills development to incorporation of a systems context, reflecting redefined values and interventions associated with safety, quality, and professional nursing roles. A course revision, based on the QSEN competencies definitions, selected beginning KSAs, exemplar resources, and teaching strategies, is described. The reframing of fundamentals of nursing is essential to prepare new graduates for contemporary practice. PMID- 20000252 TI - Embracing quality and safety education for the 21st century: building interprofessional education. AB - The education of health professions students is rooted historically in time honored and silo-bound traditions of pedagogy and content not easily influenced by outside forces. However, the quality chasm work of the Institute of Medicine, Institute of Healthcare Improvement, Quality and Safety Education for Nurses, and other groups has led to a remarkable willingness to change at one academic health sciences university. This article describes one university's strategies, challenges, and successes in delivering interprofessional educational programs. Four interprofessional learning activities, developed using a plan-do-study-act model and focused on teamwork, quality, and safety, are presented. Challenges and successes encountered are described as well as a curricular framework to enhance sustainability. PMID- 20000253 TI - Facilitating a culture of safety and patient-centered care through use of a clinical assessment tool in undergraduate nursing education. AB - Although delivery of safe, patient-centered care has long been a priority among nursing educators, nursing students may not be able to adequately translate concepts learned within the classroom into nursing practice. Nurse educators must develop teaching strategies to provide nursing students with the knowledge, skills, and attitudes necessary to provide quality patient care. Within the Quality and Safety Education for Nurses project, a set of key nursing competencies was established. This article describes the development and use of a clinical assessment tool in undergraduate clinical nursing courses to enhance mastery of two of these key nursing competencies: safety and patient-centered care. PMID- 20000254 TI - Impact of a patient safety curriculum for nurse anesthesia students. AB - Patient safety has become an important aspect of national health care initiatives. The purpose of this evaluation was to measure the impact of a patient safety education series for students enrolled in a nurse anesthesia program. Baseline surveys that measured patient safety competencies across three domains, attitudes, skills and knowledge, were administered to the students. A patient safety education series was delivered to the cohort and the survey was then readministered. Mean scores were compared using independent samples t tests. Attitude scores did not change from baseline to posttest. Participants scored higher on posttest means for both the patient safety skills and knowledge domains. Incorporating patient safety content into the nurse anesthesia master's degree curriculum may enhance clinicians' skills and knowledge related to patient safety, and the addition of a patient safety curriculum is important during the formative education process. PMID- 20000255 TI - Developing patient-centered care competencies among prelicensure nursing students using simulation. AB - Health care improvement requires collaboration between academia and practice to bridge gaps in nurse education and achievement of quality outcomes. Quality and Safety Education for Nurses identified six domains, including patient-centered care, that should be addressed during prelicensure education. Simulation is a recommended strategy to teach safe clinical practice; however, cost, space, and faculty resources are barriers to its use. Computer-based social simulation is less resource intensive and effective in improving critical-thinking skills. A pilot study was conducted to compare required resources and student learning outcomes for traditional versus computer-based simulation. Fifty-three baccalaureate nursing students participated in the study; 34 completed the computer-based simulation and 31 completed the traditional simulation. Group patient-centered care competency scores improved similarly (p < 0.001), although fewer faculty hours were required to administer the computer-based intervention. Findings suggest computer-based simulation is an efficient and effective learning strategy to develop patient-centered care competencies. PMID- 20000256 TI - Enhancing quality and safety competency development at the unit level: an initial evaluation of student learning and clinical teaching on dedicated education units. AB - The need to attend to quality and safety competency development, increase capacity in nursing education programs, address the faculty and nursing shortages, and find new ways to keep step with an ever-changing health care environment has brought forth numerous creative curricular responses and collaborative efforts. To tackle these multiple needs and challenges simultaneously, a new academic-service partnership was created to collaboratively develop an innovative clinical education delivery model. The designed dedicated education unit model not only promoted student learning about quality and safety competencies via unit-based projects but also supported quality improvements in nursing care delivery. Following the initial semester of the model's implementation, a pilot study was conducted. The findings generated the evidence required to take this innovation to the next level. Moreover, the education practice partnership, which was created to implement the clinical education delivery model, was strengthened as a result of this preliminary evaluation. PMID- 20000257 TI - Innovative learning activity. PMID- 20000258 TI - Let's call it "doctoral" advanced practice nursing. PMID- 20000259 TI - Connecting the dots of evaluation. AB - This column presents an overview of Kirkpatrick's four levels of evaluation that can be helpful to continuing nursing education providers when building evaluation into their provider units and activities. PMID- 20000260 TI - Wikis and blogs: consider the possibilities for continuing nursing education. AB - Wikis and blogs are software programs that promote communication and collaboration through the development of written documents. Nurse educators can use them to facilitate teaching and learning in a variety of health care settings. PMID- 20000261 TI - An evidence-based nurse residency program in geropalliative care. AB - As U.S. baby boomers age and the older population doubles by 2030, unprecedented numbers of older adults will need and benefit from nursing care that is evidence based and tailored to their needs and preferences. To address this need, Massachusetts General Hospital developed the RN Residency: Transitioning to Geriatrics and Palliative Care program. Guided by two national curricula, the RN Residency program was designed to expand the disease model of geriatrics to a functional, emotional, and social perspective of aging, in which palliative care is an integral component. This article describes the RN Residency program, which was designed to improve the effectiveness of nursing assessment, interventions, and outcomes for older adults and their families. PMID- 20000263 TI - The professional portfolio: an evidence-based assessment method. AB - Competency assessment is critical for a myriad of disciplines, including medicine, law, education, and nursing. Many nurse managers and educators are responsible for nursing competency assessment, and assessment results are often used for annual reviews, promotions, and satisfying accrediting agencies' requirements. Credentialing bodies continually seek methods to measure and document the continuing competence of licensees or certificants. Many methods and frameworks for continued competency assessment exist. The portfolio process is one method to validate personal and professional accomplishments in an interactive, multidimensional manner. This article illustrates how portfolios can be used to assess competence. One specialty nursing certification board's process of creating an evidence-based portfolio for recertification or reactivation of a credential is used as an example. The theoretical background, development process, implementation, and future implications may serve as a template for other organizations in developing their own portfolio models. PMID- 20000264 TI - Perceptions of select registered nurses of the continuing competence program of the Saskatchewan registered nurses' association. AB - Nursing is a self-regulating profession, and most professional nursing jurisdictions across Canada have undertaken the creation of Continuing Competence Programs (CCPs), with the goals of promoting good nursing practice, encouraging continuous learning, contributing to the quality of nursing practice, and optimizing client outcomes. Most CCPs call for a professional portfolio to collect, synthesize, and analyze professional experiences, including documentation of peer feedback and preparation of a learning plan. In the province of Saskatchewan, Canada, there is a self-reflective tool that enables registered nurses to self-rate their achievement of a set of foundational competencies. This article explores the perceptions that select registered nurses have about the CCP in Saskatchewan and how their view of the degree of professional control (as assessed through locus of control) that they have affects their perspective about the outcomes of the program. The study was designed to assess how perceived locus of control was related to how registered nurses view the implementation of the CCP in Saskatchewan. PMID- 20000265 TI - Code Carnivals: resuscitating Code Blue training with accelerated learning. AB - Nurses in the hospital setting must be knowledgeable about resuscitation procedures and proficient in the delivery of care during an emergency. They must be ready to implement their knowledge and skills at a moment's notice. A common dilemma for many nurses is that cardiopulmonary emergencies (Code Blues) are infrequent occurrences. Therefore, how do nurses remain competent and confident in their implementation of emergency skills while having limited exposure to the equipment and minimal experience in emergency situations? A team of nurse educators at a regional medical center in Washington State applied adult learning theory and accelerated learning techniques to develop and present a series of learning activities to enhance the staff's familiarity with emergency equipment and procedures. The series began with a carnival venue that provided hands-on practice and review of emergency skills and was reinforced with subsequent random unannounced code drills led by both educators and charge nurses. PMID- 20000266 TI - Preparation of nurses who precept baccalaureate nursing students: a descriptive study. AB - Clinical preceptorship is employed by baccalaureate nursing education programs as a capstone experience and a means of initiating the professional socialization of senior-level students who will shortly be new graduates. This quantitative, descriptive study explored the perceptions about preceptor preparation among nurses who precept baccalaureate nursing students. Mercer's Role Attainment Theory was the theoretical framework for this study, in which 75 participants from two hospitals in the Midwest completed a survey. Two key findings emerged from the data: All preceptors of senior-level nursing students want to know what their responsibilities are with regard to the student and the preceptorship experience, and nurses in critical care areas identified learning how to teach critical thinking to senior-level students as more essential than did nurses in other areas. PMID- 20000267 TI - Preparing clinical nurse leaders in a regional Australian teaching hospital. AB - The need to develop nurses as managers and leaders is crucial to the retention of registered nurses at a time of work force shortages and an increasingly aging work force in most Western industrialized countries. This article describes a creative and collaborative educational initiative developed at a large regional teaching hospital in New South Wales, Australia, designed to address this need. Based on a competency assessment process designed around face-to-face education, resource materials, and individualized mentoring from nurse unit managers, the aim of this multifaceted educational program is to develop effective team leaders in the clinical setting as well as a new generation of nursing leaders. PMID- 20000268 TI - Transitional care. PMID- 20000271 TI - Cultural understanding. PMID- 20000273 TI - Collaboration and the process of professional development. PMID- 20000277 TI - Responding to restraint use. PMID- 20000278 TI - What makes a generic medication generic? AB - By law, the U.S. Food and Drug Administration (FDA) can approve generic versions of brand-name drugs without requiring that research be conducted to prove them safe and effective. However, compared with a brand-name drug, a generic drug must demonstrate that it (a) contains the same active ingredients; (b) is identical in strength, dosage form, and route of administration; (c) is bioequivalent; (d) has the same clinical use indications; (e) meets the same batch requirements for identity, strength, purity, and quality; and (f) is manufactured according to the same FDA regulations. The most important aspect of generic drug development are the concepts of bioavailability and bioequivalence. According to the concept of bioequivalence, if a drug product contains an active ingredient that is chemically identical and is delivered to the site of action at the same rate and extent as another drug product, then it is considered to be clinically equivalent and can be substituted for that drug product. PMID- 20000279 TI - Pediatric bipolar disorder. AB - Pediatric bipolar disorder differs from the adult form of the disorder, marked by longer episodes, rapid cycling, prominent irritability, and high rates of comorbid attention-deficit/hyperactivity disorder and anxiety disorders. A careful assessment by families of children's symptoms, including their duration and intensity, helps with accurate diagnosis. After the diagnosis is made and careful psychopharmacological intervention is initiated, psychiatric nursing treatment of children and adolescents with pediatric bipolar disorder should involve child-and family-focused cognitive-behavioral therapies, family support, and psychoeducation. PMID- 20000280 TI - Assessing resilience. AB - As more studies reveal the positive relationships between resilience and aging well, mental health practitioners are increasingly interested in recognizing and strengthening resilience among aging Baby Boomers. Although there is much information on characteristics of resilience, there is very little research on assessing for resilience. This article presents a framework for assessing resilience among middle-aged and older adults that includes the Resilience Scale and open-ended questions to provide clients opportunities to reflect on and perhaps rediscover their resilience. This assessment framework may be useful for mental health practitioners using a strengths-based approach with their clients. PMID- 20000281 TI - Dealing with bullying in the workplace: toward zero tolerance. AB - Recent reports indicate an absence of respect in workplace culture. Every person has the right to a workplace that is fair and equitable and in which he or she is treated with respect and dignity. Working in a collegial manner is challenging when environments have staff who engage in unprofessional or disruptive behaviors. A number of steps can be taken to support a healthy workplace and prevent bullying. Healthy workplaces are ones in which leaders and managers lead by example, champion respect, and set the tone and expectation for behaviors essential for fostering a harmonious and collaborative environment. The role of the leader/manager is crucial in developing a positive workplace culture that supports a high level of professionalism and a culture of zero tolerance toward bullying. PMID- 20000282 TI - The effect of animal-assisted activity on inpatients with schizophrenia. AB - The aim of this study was to evaluate the effects of animal-assisted activity on self-esteem, control over activities of daily living, and other psycho physiological aspects among Taiwanese inpatients with schizophrenia. Thirty participants were randomly assigned to either the treatment or control group. A weekly animal-assisted activity program was arranged for patients in the treatment group for 2 months. A questionnaire assessing self-esteem, self determination, social support, and psychiatric symptoms was completed the week before and the week after the animal-assisted activity. Compared with the control group, the treatment group showed significant improvement on all measures except for social support and negative psychiatric symptoms. The results of this study showed that animal-assisted activity can promote significant improvements in many clinical aspects among inpatients with schizophrenia. Therefore, animal-assisted activity should be integrated into the treatment of institutionalized patients with schizophrenia. PMID- 20000284 TI - Remodeling effect of cross-linking. PMID- 20000285 TI - Preoperative keratometry and visual outcomes after hyperopic LASIK. PMID- 20000286 TI - Initial clinical evaluation of an intraocular femtosecond laser in cataract surgery. AB - PURPOSE: To evaluate femtosecond laser lens fragmentation and anterior capsulotomy in cataract surgery. METHODS: Anterior capsulotomy and phacofragmentation procedures performed with an intraocular femtosecond laser (LenSx Lasers Inc) were initially evaluated in ex vivo porcine eyes. These procedures were then performed in an initial series of nine patients undergoing cataract surgery. In addition to standard intraoperative assessments (including capsulotomy diameter accuracy and reproducibility), optical coherence tomography was used to evaluate human procedures. RESULTS: For an intended 5-mm capsulorrhexis in porcine eyes, average achieved diameters were 5.88+/-0.73 mm using a standard manual technique and 5.02+/-0.04 mm using the femtosecond laser. Scanning electron microscopy revealed equally smooth cut edges of the capsulotomy with the femtosecond laser and manual technique. Compared to control porcine eyes, femtosecond laser phacofragmentation resulted in a 43% reduction in phacoemulsification power and a 51% decrease in phacoemulsification time. In a small series of human clinical procedures, femtosecond laser capsulotomies and phacofragmentation demonstrated similarly high levels of accuracy and effectiveness, with no operative complications. CONCLUSIONS: Initial results with an intraocular femtosecond laser demonstrate higher precision of capsulorrhexis and reduced phacoemulsification power in porcine and human eyes. PMID- 20000287 TI - Calculation of intraocular lens power using Orbscan II quantitative area topography after corneal refractive surgery. AB - PURPOSE: To present the prospective application of the Orbscan II central 2-mm total-mean corneal power obtained by quantitative area topography in intraocular lens (IOL) calculation after refractive surgery. METHODS: Calculated and achieved refraction and the difference between them were studied in 77 eyes of 61 patients with previous radial keratotomy (RK), RK and additional surgeries, myopic LASIK, myopic photorefractive keratectomy (PRK), or hyperopic LASIK who underwent phacoemulsification without complications in 3 eye centers. All IOL calculations used the average from the central 2-mm Orbscan II total-mean power of maps centered on the pupil without the use of previous refractive data. Six IOL styles implanted within the bag were used. RESULTS: Using the SRK-T formula, the overall calculated refraction was -0.64+/-0.93 diopters (D). The overall achieved spherical equivalent refraction (-0.52+/-0.79 D; range: -3.12 to 1.25 D; 95% confidence interval [CI]: -0.70/-0.34 D) was +/-0.50 D in 53% of eyes, +/-1.00 D in 78% of eyes, and +/-2.00 D in 99% of eyes. The overall difference between the calculated and achieved refraction (0.12+/-0.93 D, P=.27; range: -2.18 to 2.62 D; 95% CI: 0.09/0.33 D) was +/-0.50 D in 39% of eyes, +/-1.00 D in 77% of eyes, and +/-2.00 D in 96% of eyes. This difference was +/-1.00 D in 77% of eyes with RK (P=.70), 82% of eyes with myopic LASIK (P=.34), and 90% of eyes with myopic PRK (P=.96). In eyes with RK followed by LASIK, a trend toward undercorrection was noted (P=.03). In eyes with hyperopic LASIK, a trend toward overcorrection was noted (P=.005). CONCLUSIONS: In eyes with previous corneal refractive surgery, IOL power calculation can be performed with reasonable accuracy using the Orbscan II central 2-mm total-mean power. This method had better outcomes in eyes with previous RK, myopic LASIK, and myopic PRK than in eyes with hyperopic LASIK or RK with LASIK. PMID- 20000288 TI - Vector analysis of cross cylinder LASIK with the NIDEK EC-5000 excimer laser for high astigmatism. AB - PURPOSE: To evaluate visual outcomes of LASIK in eyes with high astigmatic refractive errors with low spherical component. METHODS: We prospectively evaluated cross cylinder LASIK in patients with high astigmatic eyes with low spherical component over a 2-year period. All laser surgeries were performed with the NIDEK EC-5000 excimer laser. We used power vectors (M [spherical equivalent] and J(0), and J(45) [cylinder]) to analyze the astigmatic results. Main outcome measures were uncorrected visual acuity (UCVA), best spectacle-corrected visual acuity (BSCVA), manifest refraction, and complications. RESULTS: Thirty-four eyes of 22 patients were included. Patients were followed for 21.8+/-3.7 months. At the last examination, UCVA of 20/40 or better was observed in 33 (97.1%) eyes. Best spectacle-corrected visual acuity after LASIK improved significantly in comparison to BSCVA before surgery (P<.005). Mean preoperative cylinder was 4.73+/-0.89 diopters (D) (range: -4.00 to -7.00 D), which decreased to -0.29+/ 0.47 D (range: 0.00 to -1.50 D) at last follow-up (P<.001). Reduction in the magnitude of power vectors was 92.4% for M, 103.2% for J(0), and 76.4% for J(45). No cases of post-operative astigmatic regression or corneal haze were observed. CONCLUSIONS: Cross cylinder LASIK with the NIDEK EC-5000 excimer laser is a successful procedure for correcting high astigmatism and improving vision. PMID- 20000289 TI - Clinical outcomes of LASIK for myopia using the SCHWIND platform with ocular wavefront customized ablation. AB - PURPOSE: To evaluate the clinical outcomes of aspheric ocular wavefront ablation profiles in LASIK treatments. METHODS: Thirty eyes treated by ocular wavefront were retrospectively analyzed at 6-month follow-up. Custom Ablation Manager (CAM) software was used to plan wavefront-customized aspheric treatments, and the ESIRIS system was used to perform ablations (SCHWIND eye-tech-solutions). Outcomes were evaluated in terms of efficacy, predictability, refractive outcome, safety, and pre-and postoperative wavefront aberration analysis (SCHWIND Ocular Wavefront Analyzer). RESULTS: At 6 months postoperatively, 47% of eyes achieved uncorrected visual acuity 20/20 or better. Average defocus was reduced from 3.49+/-2.38 diopters (D) preoperatively (range: -10.63 to 0.00 D) to -0.14+/-0.31 D postoperatively (range: -1.75 to 0.00 D). Astigmatism was reduced from -0.81+/ 1.15 D (range: -4.25 to 0.00 D) to -0.25+/-0.37 D (range: -1.25 to 0.00 D). Eighty-six percent of eyes were within +/- 0.50 D. Best spectacle-corrected visual acuity (logMAR) improved from +0.12+/-0.08 (range: 0.0 to +0.2) to -0.05+/ 0.13 (range: -0.2 to +0.2) (P=.04). The treatment did not change coma or spherical aberration, and reduced the trefoil from 0.21+/-0.13 microm (range: 0.05 to 0.53 microm) to 0.08+/-0.13 microm (range: 0.01 to 0.39 microm) (P=.002). CONCLUSIONS: The study results indicate that the aspheric ocular wavefront customized CAM approach for planning ablation volumes is safe and effective. PMID- 20000290 TI - Anterior segment measurements using Pentacam and Orbscan II 1 to 5 years after refractive surgery. AB - PURPOSE: To compare anterior segment measurements between Pentacam and Orbscan II after laser surface ablation. METHODS: Corneal thickness, anterior elevation, posterior elevation, and anterior chamber depth more than 1 year after laser refractive surgery were measured using Pentacam and Orbscan II in 51 eyes that underwent laser epithelial keratomileusis (LASEK) and 21 eyes that underwent photorefractive keratectomy (PRK). As controls, the same comparisons were made in 52 unoperated eyes. RESULTS: Mean follow-up was 28.2 months in the LASEK group and 49.9 months in the PRK group. Mean corneal thickness using Pentacam was thicker than that using Orbscan by 10.4 microm in unoperated eyes, 22.6 microm in LASEK eyes, and 11.0 microm in PRK eyes, but the two devices showed good correlation in all groups. Mean anterior elevation was highly correlated between the two devices with 2.1-microm differences in LASEK eyes, but posterior elevation showed no correlation with significant discrepancies. Mean anterior chamber depth was 0.12-to 0.14-microm thicker with Pentacam than with Orbscan, and highly correlated in all groups. CONCLUSIONS: Pentacam and Orbscan showed good correlations in corneal thickness, anterior elevation, and anterior chamber depth despite some differences in mean values. The two devices, however, showed no correlation in posterior elevation measurements, a difference that was greater in the postoperative groups. PMID- 20000291 TI - Corneal endothelial cell density after femtosecond thin-flap LASIK and PRK for myopia: a contralateral eye study. AB - PURPOSE: To compare the effect of femtosecond thinflap LASIK and photorefractive keratectomy (PRK) on postoperative endothelial cell density. METHODS: In a prospective, randomized, contralateral, single-center clinical trial, 25 patients (mean age: 30+/-5 years [range: 21 to 38 years]) underwent PRK in one eye and thin-flap LASIK in the fellow eye for the correction of myopia using a wavefront guided platform. The central corneal endothelial cell density was measured using the NIDEK Confoscan 4 preoperatively, and at 1 and 3 months postoperatively. Changes in endothelial cell density were analyzed over time between the two refractive techniques. RESULTS: In PRK, the average preoperative endothelial cell density was 3011+/-329 cells/mm(2), which decreased to 2951+/-327 cells/mm(2) at 1 month (P=.5736) and 2982+/-365 cells/mm(2) at 3 months (P=.6513). In thinflap LASIK, the average preoperative endothelial cell density was 2995+/-325 cells/mm(2), which decreased to 2977+/-358 cells/mm(2) at 1 month (P=.5756) and 2931+/-369 cells/mm(2) at 3 months (P=.4106). No statistically significant difference was found between the two groups at 1 (P=.7404) or 3 (P=.3208) months postoperatively. CONCLUSIONS: No statistically significant change was noted in endothelial cell density following either PRK or thin-flap LASIK for the treatment of myopia. Furthermore, no statistically significant difference was found between the two groups out to 3 months postoperatively, indicating that thin-flap LASIK is as safe as PRK with regards to endothelial health. PMID- 20000292 TI - Outcomes of the Acrysof ReSTOR IOL in myopes, emmetropes, and hyperopes. AB - PURPOSE: To evaluate the influence of preoperative ocular parameters on various parameters of postoperative outcome after implantation of the AcrySof ReSTOR intraocular lens (IOL) in myopic, hyperopic, and emmetropic patients. METHODS: Phacoemulsification and IOL implantation was performed in 56 eyes of 32 patients. According to axial length, 28 eyes were classified as emmetropic, 16 as myopic, and 12 as hyperopic. Patients were examined after 8.2+/-1.8 months (range: 5.7 to 10.5 months) for uncorrected and best distance-corrected visual acuity for far, intermediate, and near vision. A defocus profile was recorded and individual reading distance and distance for lowest intermediate visual acuity were determined. The predictability of three different calculation formulas was evaluated. Patients were interviewed for photic phenomena, quality of night vision, spectacle independence, and satisfaction with visual outcome. RESULTS: Predictability of IOL calculation was not statistically significant among groups using the SRK-T formula, but was significantly different with the Haigis (P=.01) or Holladay I (P=.02) formulas. Uncorrected and best distance-corrected visual acuity for near, intermediate, and far vision were not significantly different among groups. Mean uncorrected far visual acuity was 20/23+/-5.5 letters, 20/25+/ 6 letters, and 20/25+/-4.5 letters and uncorrected near visual acuity was 20/25+/ 5.5 letters, 20/26+/-4.5 letters, and 20/26+/-7.5 letters for the emmetropic, myopic, and hyperopic groups, respectively. The defocus profile significantly varied among groups; patients of the hyperopic group showed a closer reading distance (29.5 cm) than patients of the emmetropic (32.8 cm) and myopic (34.6 cm) groups. Reading distance was statistically significantly correlated to axial length and anterior chamber depth and thus can be estimated preoperatively. CONCLUSIONS: Our data show that preoperative refractive status is a relevant factor influencing postoperative outcome after implantation of a ReSTOR IOL. Predictability of IOL calculation and defocus profile were significantly different for myopic, emmetropic, and hyperopic patients. PMID- 20000293 TI - Wavefront-guided photorefractive keratectomy to correct ametropia following aspheric ReSTOR implantation. AB - PURPOSE: To describe the clinical course of three eyes of two patients who underwent wavefront-guided photorefractive keratectomy to correct ametropia following cataract extraction and aspheric ReSTOR lens (SN6AD3; +4.00-diopter model) implantation. METHODS: Pre-and postoperative uncorrected visual acuity (UCVA) and best spectacle-corrected visual acuity (BSCVA), manifest refraction, and WaveScans based on the Hartmann-Shack analyzer were evaluated. The VISX STAR S4 excimer with eye tracking and iris registration was used in all cases. RESULTS: Good quality WaveScans were obtained; UCVA improved in all cases and BSCVA improved or remained unchanged. CONCLUSIONS: These cases demonstrate that it is possible to obtain good quality wavefront data in patients with the ReSTOR lens and to use such data to design wavefront-guided treatment to maximize uncorrected distance and near visual acuities after ReSTOR implantation. PMID- 20000296 TI - Treatment of diabetic foot ulcers with tendon lengthening. PMID- 20000297 TI - Delivery options for prevention of perinatal HIV transmission. PMID- 20000299 TI - NSAIDs and cardiovascular risk. PMID- 20000300 TI - NSAID prescribing precautions. AB - Nonsteroidal anti-inflammatory drugs (NSAIDs) are commonly used, but have risks associated with their use, including significant upper gastrointestinal tract bleeding. Older persons, persons taking anticoagulants, and persons with a history of upper gastrointestinal tract bleeding associated with NSAIDs are at especially high risk. Although aspirin is cardioprotective, other NSAIDs can worsen congestive heart failure, can increase blood pressure, and are related to adverse cardiovascular events, such as myocardial infarction and ischemia. Cyclooxygenase-2 inhibitors have been associated with increased risk of myocardial infarction; however, the only cyclooxygenase-2 inhibitor still available in the United States, celecoxib, seems to be safer in this regard. Hepatic damage from NSAIDs is rare, but these medications should not be used in persons with cirrhotic liver diseases because bleeding problems and renal failure are more likely. Care should be used when prescribing NSAIDs in persons taking anticoagulants and in those with platelet dysfunction, as well as immediately before surgery. Potential central nervous system effects include aseptic meningitis, psychosis, and tinnitus. Asthma may be induced or exacerbated by NSAIDs. Although most NSAIDs are likely safe in pregnancy, they should be avoided in the last six to eight weeks of pregnancy to prevent prolonged gestation from inhibition of prostaglandin synthesis, premature closure of the ductus arteriosus, and maternal and fetal complications from antiplatelet activity. Ibuprofen, indomethacin, and naproxen are safe in breastfeeding women. Care should be taken to prevent accidental NSAID overdose in children by educating parents about correct dosing and storage in childproof containers. PMID- 20000301 TI - Intrapartum fetal monitoring. AB - Continuous electronic fetal monitoring was developed in the 1960s to assist in the diagnosis of fetal hypoxia during labor. Continuous electronic fetal monitoring has been shown to reduce the incidence of neonatal seizures, but there has been no beneficial effect in decreasing cerebral palsy or neonatal Continuous electronic fetal monitoring was developed in the 1960s to assist in the diagnosis of fetal hypoxia during labor. Continuous electronic fetal monitoring has been shown to reduce the incidence of neonatal seizures, but there has been no beneficial effect in decreasing cerebral palsy or neonatal mortality. Intraobserver variability may play a major role in its interpretation. To provide a systematic approach to interpreting the electronic fetal monitor tracing, the National Institute of Child Health and Human Development convened a workshop in 2008 to revise the accepted definitions for electronic fetal monitor tracing. The key elements include assessment of baseline heart rate, presence or absence of variability, and interpretation of periodic changes. The workshop introduced a new classification scheme for decision making with regard to tracings. This system can be used in conjunction with the Advanced Life Support in Obstetrics course mnemonic, DR C BRAVADO, to assist in the systematic interpretation of fetal monitoring. DR C BRAVADO incorporates maternal and fetal risk factors (DR = determine risk), contractions (C), the fetal monitor strip (BRA = baseline rate, V = variability, A = accelerations, and D = decelerations), and interpretation (O = overall assessment). PMID- 20000302 TI - Evening primrose oil. AB - Evening primrose oil (Oenothera biennis) is a commonly used alternative therapy and a rich source of omega-6 essential fatty acids. It is best known for its use in the treatment of systemic diseases marked by chronic inflammation, such as atopic dermatitis and rheumatoid arthritis. It is often used for several women's health conditions, including breast pain (mastalgia), menopausal and premenstrual symptoms, cervical ripening, and labor induction or augmentation. However, there is insufficient evidence to make a reliable assessment of its effectiveness for most clinical indications. The current evidence suggests that oral evening primrose oil does not provide clinically significant improvement in persons with atopic dermatitis, and that it is also likely ineffective for the treatment of cyclical mastalgia and premenstrual syndrome. However, most trials to date have significant methodologic flaws and must be considered preliminary. The use of evening primrose oil during pregnancy is not supported in the literature and should be avoided. Evening primrose oil is generally well tolerated, with reported minor adverse effects, including gastrointestinal upset and headaches. Optimal dosing standards and treatment regimens await clarification in adequately powered clinical trials. PMID- 20000303 TI - Latex allergy. AB - The prevalence of latex allergy in the general population is low; however, the risk of developing latex allergy is higher in persons with increased latex exposure, such as health care workers or persons who work in the rubber industry. Children with spina bifida and others who undergo multiple surgeries or procedures, particularly within the first year of life, are also at greater risk of latex allergy. Reactions to latex allergy can range from type IV delayed hypersensitivity (e.g., contact dermatitis) to type I immediate hypersensitivity (e.g., urticaria, bronchospasm, anaphylaxis). Latex allergy can be diagnosed with clinical history, skin prick testing, latex-specific serum immunoglobulin E testing, and glove provocation testing. The main goals of latex allergy management are avoidance of exposure to latex allergens and appropriate treatment of allergic reactions. The use of nonlatex products from birth may prevent potentially serious allergic reactions. Widespread adoption of nonlatex or low latex gloves has decreased the incidence of latex sensitization in health care workers. PMID- 20000304 TI - Evaluation and treatment of hematospermia. AB - Hematospermia can be a distressing symptom for patients, but most cases are effectively managed by a primary care physician. Although the condition is usually benign, significant underlying pathology must be excluded by history, physical examination, laboratory evaluation, and, in select cases, other diagnostic modalities. In men younger than 40 years without risk factors (e.g., history of cancer, known urogenital malformation, bleeding disorders) and in men with no associated symptoms, hematospermia is often self-limited and requires no further evaluation or treatment other than patient reassurance. Many cases are attributable to sexually transmitted infections or other urogenital infections in men younger than 40 years who present with hematospermia associated with lower urinary tract symptoms. Workup in these patients can be limited to urinalysis and testing for sexually transmitted infections, with treatment as indicated. In men 40 years and older, iatrogenic hematospermia from urogenital instrumentation or prostate biopsy is the most common cause of blood in the semen. However, recurrent or persistent hematospermia or associated symptoms (e.g., fever, chills, weight loss, bone pain) should prompt further investigation, starting with a prostate examination and prostate-specific antigen testing to evaluate for prostate cancer. Other etiologies to consider in those 40 years and older include genitourinary infections, inflammations, vascular malformations, stones, tumors, and systemic disorders that increase bleeding risk. PMID- 20000305 TI - Hip impingement: identifying and treating a common cause of hip pain. AB - Femoroacetabular impingement, also known as hip impingement, is the abutment of the acetabular rim and the proximal femur. Hip impingement is increasingly recognized as a common etiology of hip pain in athletes, adolescents, and adults. It injures the labrum and articular cartilage, and can lead to osteoarthritis of the hip if left untreated. Patients with hip impingement often report anterolateral hip pain. Common aggravating activities include prolonged sitting, leaning forward, getting in or out of a car, and pivoting in sports. The use of flexion, adduction, and internal rotation of the supine hip typically reproduces the pain. Radiography, magnetic resonance arthrography, and injection of local anesthetic into the hip joint confirm the diagnosis. Pain may improve with physical therapy. Treatment often requires arthroscopy, which typically allows patients to resume premorbid physical activities. An important goal of arthroscopy is preservation of the hip joint. Whether arthroscopic treatment prevents or delays osteoarthritis of the hip is unknown. PMID- 20000306 TI - Applying HIV testing guidelines in clinical practice. AB - An estimated one fourth of persons with human immunodeficiency virus (HIV) are not aware they are infected. Early diagnosis of HIV has the potential to ensure optimal outcomes for infected persons and to limit the spread of the virus. Important barriers to testing among physicians include insufficient time, reimbursement issues, and lack of patient acceptance. Current HIV testing guidelines address many of these barriers by making the testing process more streamlined and less stigmatizing. The opt-out consent process has been shown to improve test acceptance. Formal pretest counseling and written consent are no longer recommended by the Centers for Disease Control and Prevention. Nevertheless, pretest discussions provide an opportunity to give information about HIV, address fears of discrimination, and identify ongoing high-risk activities. With increased HIV screening in the primary care setting, more persons with HIV could be identified earlier, receive timely and appropriate care, and get treatment to prevent clinical progression and transmission. PMID- 20000310 TI - FPIN's Clinical Inquiries. Aspirin use in children for fever or viral syndromes. AB - Aspirin should not be used to treat acute febrile viral illness in children. (Strength of Recommendation [SOR]: C, based on case-control studies). Although no causal link has been proven, data from case-control and historic cohort studies demonstrate an association between aspirin use and Reye syndrome. The risk of Reye syndrome decreases with age, becoming extremely rare by the late teenage years. Other nonsteroidal anti-inflammatory drugs are effective antipyretics and are not associated with the constellation of symptoms seen in Reye syndrome, which includes nausea, vomiting, headache, excitability, delirium, combativeness, and coma. Aspirin use in children younger than 19 years should be limited to diseases in which aspirin has a proven benefit, such as Kawasaki disease and the juvenile arthritides. (SOR: C, based on expert opinion). PMID- 20000311 TI - Nonhealing diaper rash with associated hepatosplenomegaly. PMID- 20000312 TI - Cutaneous abdominal nodule after cesarean delivery. PMID- 20000314 TI - Extraction and characterization of latex and natural rubber from rubber-bearing plants. AB - Consecutive extraction of latex and natural rubber from the roots of rubber bearing plants such as Taraxacum kok-saghyz (TKS), Scorzonera tau-saghyz (STS), and Scorzonera Uzbekistanica (SU) were carried out. Latex extraction was carried via two methods: Blender method and Flow method. The results of latex extraction were compared. Cultivated rubber-bearing plants contained slightly higher latex contents compared to those from wild fields. Several creaming agents for latex extraction were compared. About 50% of total natural rubber was extracted as latex. The results of the comparative studies indicated that optimum latex extraction can be achieved with Flow method. The purity of latex extracted by Blender method ( approximately 75%) was significantly lower than that extracted by Flow method (99.5%). When the latex particles were stabilized with casein, the latex was concentrated significantly. Through concentrating latex by flotation, the latex concentration of 35% was obtained. Bagasse contained mostly solid natural rubber. The remaining natural rubber in the bagasse (left after the latex extraction) was extracted using sequential solvent extraction first with acetone and then with several nonpolar solvents. Solid natural rubber was analyzed for gel content and characterized by size exclusion chromatography (SEC) for molecular weight determinations. SEC of solid natural rubber has shown that the molecular weight is about 1.8E6 and they contain less gel compared to TSR20 (Grade 20 Technically Specified Rubber), a commercial natural rubber from Hevea brasiliensis. PMID- 20000315 TI - Dissolution potential of SO2 Co-injected with CO2 in geologic sequestration. AB - Sulfur dioxide is a possible co-injectant with carbon dioxide in the context of geologic sequestration. Because of the potential of SO2 to acidify formation brines, the extent of SO2 dissolution from the CO2 phase will determine the viability of co-injection. Pressure-, temperature-, and salinity-adjusted values of the SO2 Henry's Law constant and fugacity coefficient were determined. They are predicted to decrease with depth, such that the solubility of SO2 is a factor of 0.04 smaller than would be predicted without these adjustments. To explore the potential effects of transport limitations, a nonsteady-state model of SO2 diffusion through a stationary cone-shaped plume of supercritical CO2 was developed. This model represents an end-member scenario of diffusion-controlled dissolution of SO2, to contrast with models of complete phase equilibrium. Simulations for conditions corresponding to storage depths of 0.8-2.4 km revealed that after 1000 years, 65-75% of the SO2 remains in the CO2 phase. This slow release of SO2 would largely mitigate its impact on brine pH. Furthermore, small amounts of SO2 are predicted to have a negligible effect on the critical point of CO2 but will increase phase density by as much as 12% for mixtures containing 5% SO2. PMID- 20000316 TI - Structural phase transitions in EuC(2). AB - Pure EuC(2), free of EuO impurities, was obtained by the reaction of elemental europium with graphite at 1673 K. By means of synchrotron powder diffraction experiments, the structural behavior was investigated in the temperature range from 10 to 1073 K. In contrast to former results, EuC(2) crystallizes in the ThC(2) type structure (C2/c, Z = 4) at room temperature. A tetragonal modification (I4/mmm, Z = 2) is only observed in a very small temperature range just below the transition to a cubic high-temperature modification (Fm3m, Z = 4) at 648 K. DTA/TG investigations confirm these results. According to Raman spectroscopy, EuC(2) contains C(2)(2-) ions (nu(C[triple bond]C) = 1837 cm(-1)). The divalent character of Eu is confirmed by the results of magnetic susceptibility measurements and (151)Eu Mossbauer spectroscopy. In these measurements a transition to a ferromagnetic state with T(C) = 15 K is observed, which is in reasonable agreement with literature data. Above T(C) EuC(2) is a semiconductor according to measurements of the electric resistivity vs temperature, again in contrast to former results. Around T(C) a sharp maximum of the electric resistivity vs temperature curve was observed, which collapses on applying external magnetic fields. The observed CMR effect (colossal magnetoresistance) is much stronger than that reported for other EuC(2) samples in the literature. These investigations explicitly show the influence of sample purity on the physical and even structural properties of EuC(2). PMID- 20000317 TI - Synthesis and characterization of unsymmetric indolodithienopyrrole and extended diindolodithienopyrrole. AB - Indole-fused dithieno[3,2-b:2',3'-d]pyrrole-based unsymmetrical and extended heteroacenes were synthesized and characterized. Solid-state structures were examined using single-crystal X-ray diffraction to understand their packing behavior. The optical and electrochemical properties of these new heteroacenes are also described in detail. PMID- 20000318 TI - Size-dependent electrochemical oxidation of silver nanoparticles. AB - Here we quantify the electrochemical oxidation of Ag nanoparticles (NPs) as a function of size by electrostatically attaching Ag NPs synthesized by seed mediated growth in the presence of citrate (diameter = 8 to 50 nm) to amine functionalized indium-tin oxide coated glass electrodes (Glass/ITO), obtaining a linear sweep voltammogram from 0.1 V, where Ag(0) is stable, up to 1.0 V, and observing the peak potential (E(p)) for oxidation of Ag(0) to Ag(+). Electrostatic attachment to the organic linker presumably removes direct interactions between Ag and ITO and allows control over the total Ag coverage by altering the soaking time. This is important as both metal-electrode interactions and overall Ag coverage can affect E(p). E(p) shifts positive from an average of 275 to 382 mV as the Ag NP diameter increases for a constant Ag coverage and under conditions of planar diffusion, suggesting a shift in E(p) due to a thermodynamic shift in E(0) for the Ag/Ag(+) redox couple with size. The negative shift in E(p) with decreasing Ag NP radius follows the general trend predicted by theory and agrees with previous qualitative experimental observations. A better understanding of metal nanostructure oxidation is crucial considering their potential use in many different applications and the importance of metal corrosion processes at the nanoscale. PMID- 20000319 TI - Accurate automated protein NMR structure determination using unassigned NOESY data. AB - Conventional NMR structure determination requires nearly complete assignment of the cross peaks of a refined NOESY peak list. Depending on the size of the protein and quality of the spectral data, this can be a time-consuming manual process requiring several rounds of peak list refinement and structure determination. Programs such as Aria, CYANA, and AutoStructure can generate models using unassigned NOESY data but are very sensitive to the quality of the input peak lists and can converge to inaccurate structures if the signal-to-noise of the peak lists is low. Here, we show that models with high accuracy and reliability can be produced by combining the strengths of the high-resolution structure prediction program Rosetta with global measures of the agreement between structure models and experimental data. A first round of models generated using CS-Rosetta (Rosetta supplemented with backbone chemical shift information) are filtered on the basis of their goodness-of-fit with unassigned NOESY peak lists using the DP-score, and the best fitting models are subjected to high resolution refinement with the Rosetta rebuild-and-refine protocol. This hybrid approach uses both local backbone chemical shift and the unassigned NOESY data to direct Rosetta trajectories toward the native structure and produces more accurate models than AutoStructure/CYANA or CS-Rosetta alone, particularly when using raw unedited NOESY peak lists. We also show that when accurate manually refined NOESY peak lists are available, Rosetta refinement can consistently increase the accuracy of models generated using CYANA and AutoStructure. PMID- 20000320 TI - One-pot nucleation, growth, morphogenesis, and passivation of 1.4 nm Au nanoparticles on self-assembled rosette nanotubes. AB - A one-pot strategy for the nucleation, growth, morphogenesis, and passivation of 1.4 nm Au nanoparticles (NPs) on self-assembled rosette nanotubes (RNTs) is described. Tapping-mode atomic force microscopy, transmission electron microscopy, energy-dispersive X-ray analysis, and selected-area electron diffraction were used to establish the structure and organization of this hybrid material. Notably, we found that the Au NPs formed were nearly monodisperse clusters of Au(55) (1.4-1.5 nm) nestled in pockets on the RNT surface. PMID- 20000321 TI - Large-scale synthesis of SnO2 nanosheets with high lithium storage capacity. AB - In this communication, we successfully synthesized a new SnO(2) nanoarchitecture: extremely thin sheets, with minimum thicknesses of 1.5-3.0 nm. The products were prepared through a facile hydrothermal treatment using tin dichloride as the precursor. Planar or scrolled SnO(2) sheets were carefully examined by transmission electron microscopy. The assemblies of these sheets have a high BET surface area of 180.3 m(2)/g and extraordinarily large pore volume of 1.028 cm(3)/g. They also exhibit a high lithium storage capacity and excellent cyclability due to its nanometer-sized frame and breathable characteristic. PMID- 20000322 TI - Reduction of bis(dithiolene)oxo(disulfido)tungsten(VI) complex with dihydrogen related to the chemical function of the fourth tungsten-containing enzyme (WOR4) from Pyrococcus furiosus. AB - Sulfurization of five-coordinate [W(IV)O(1,2-benzenedithiolate)(2)](2-) proceeds under very mild conditions to form seven-coordinate [W(VI)O(eta(2)-S(2))(1,2 benzenedithiolate)(2)](2-), from which H(2)S was generated by treatment with H(2). This is the first functional model for the fourth tungsten-containing WOR4 enzyme. Kinetics and isotope-scrambling experiments have suggested the existence of six-coordinate W(V)(eta(1)-S(2)) species as a key reaction intermediate. PMID- 20000323 TI - Control of protein adsorption onto core-shell tubular and vesicular structures of diphenylalanine/parylene. AB - The self-assembly of peptides, specifically dipeptides, offers numerous advantages for biological applications. We describe an easy, versatile method of fabricating different types of zwitterionic Phe-Phe dipeptide structures (i.e., tubes and vesicles) through solvent-mediated assembly. The stability of the dipeptide structures is increased by thin polymer coatings of poly(chloro-p xylylene), a PPX film. We also investigated protein adsorption onto PPX-coated peptide tubes and vesicles by varying the thickness of the polymer film. PMID- 20000324 TI - Effect of headgroup on electrical conductivity of self-assembled monolayers on mercury: n-alkanethiols versus n-alkaneselenols. AB - The relative efficiencies of electron tunneling across self-assembled monolayers (SAMs) of n-alkanethiols and n-alkaneselenols, CH(3)-(CH(2))(n)-XH, where n = 8, 9, 11, and X = S or Se, deposited on mercury electrodes were measured via electroreduction of Ru(NH(3))(6)(3+) in aqueous solutions. Electron tunneling rates across the monolayer films decay exponentially with the monolayer thickness with a tunneling coefficient, beta = 1.1 +/- 0.1 per CH(2) irrespective of the identity of the -XH headgroup. Electron tunneling rates across n-alkanethiol monolayers are ca. 4-fold larger than the rates measured across n-alkaneselenol monolayers containing the same number of carbon atoms, signifying the importance of headgroup/metal contact resistance in electron transfer through SAMs on mercury. PMID- 20000325 TI - Porous networks of CdSe nanocrystal chains from ultrafine Cd(OH)2 nanowires and their composite materials. AB - Long ultrathin Cd(OH)(2) nanowires have been selectively grown on silica colloids in a basic aqueous condition. The Cd(OH)(2) nanowires could be detached from the surface of the silica colloids by simply applying ultrasonication and then transformed into isolated CdSe nanocrystal chains. When the transformation into CdSe was conducted without detaching the Cd(OH)(2) nanowires, nanoporous CdSe shells composed of wire-like nanocrystal chains were produced. The good solubility of the Cd(OH)(2) nanowires in both hydrophilic and hydrophobic solvents facilitated the formation of composites with quantum dots, magnetic particles, organic molecules, and polymers. Embedding premade quantum dots possessed broad light absorption range and enhanced photoluminescence. Large amount of superparamagnetic particles endowed a fast magnetic response in addition to the fluorescence. Composites of organic/nanocrystal chains were readily fabricated by employing the electrostatic attraction between the positively charged Cd(OH)(2) nanowires and negatively charged polymers or small molecules. PMID- 20000326 TI - Quantitative, label-free detection of five protein biomarkers using multiplexed arrays of silicon photonic microring resonators. AB - Because of the inherent complexity of biochemical pathways commonly altered in disease states, it has become accepted that multiplexed analyses can provide a more informative biomolecular understanding of disease onset and progression. Importantly, compared to conventional single-parameter assays, the detailed biomolecular insight gleaned from multiparameter measurements has the potential to greatly improve disease diagnostics, prognostics, and theragnostics. We have previously reported the utility of silicon photonic microring resonators for the sensitive quantitation of a single disease biomarker and herein demonstrate the first example of optical microcavity resonator arrays performing quantitative, label-free, multiplexed analyses of clinically relevant protein biomarkers. In this report, the concentrations of prostate specific antigen (PSA), alpha fetoprotein (AFP), carcinoembryonic antigen (CEA), tumor necrosis factor-alpha (TNF-alpha), and interleukin-8 (IL-8) are simultaneously determined in three unknown protein cocktail solutions. This letter demonstrates that multiple immunoassays can be performed concurrently on a microresonator platform without any accompanying loss of sensitivity or measurement precision, and therefore, this report lays the groundwork for future applications involving multiplexed analysis of clinically relevant samples. PMID- 20000327 TI - Insight into the mechanism of antimicrobial conjugated polyelectrolytes: lipid headgroup charge and membrane fluidity effects. AB - The interactions of antimicrobial cationic conjugated polyelectrolytes (CPEs) with two model membranes, liposomes and lipid monolayers at the air-water interface, have been investigated by fluorescence emission, fluorescence quenching, pressure-area isotherm, and dynamic light scattering measurements. This study continues the evaluation of the antimicrobial mechanism of a cationic poly(phenylene ethynylene) (PPE)-based CPE (polymer 1), which contains a 2,5 thienylene moiety in the repeat unit. To this end, the interactions of polymer 1 with lipids with varying headgroup charge and acyl chain length have been examined. Our results show that the cationic polymer 1 can efficiently associate with and insert into anionic phosphatidylglycerol (PG) membranes. However, polymer 1 does not exhibit any interactions with zwitterionic lipid membranes composed of phosphatidylcholine (PC) and phosphatidylethanolamine (PE) lipids. Polymer 1's selective affinity toward anionic lipids over zwitterionic lipids makes it an attractive antimicrobial agent with low toxicity. The interactions of polymer 1 with lipid membranes of different fluidity were studied by varying the surface pressure of lipid monolayers and by adjusting the temperature of liposomes. We observe that increasing membrane fluidity enhances both the conformational changes of polymer 1 upon associating with lipid membranes and the extent of polymer 1 insertion into lipid monolayers. We also find that the thickness of the lipid bilayers, modulated by acyl chain length, affects the extent of polymer 1 incorporation into the lipid bilayer. PMID- 20000328 TI - Role of hydroxyls in oxide wettability. AB - Oxides are believed to be hydrophilic because of the strong affinity for hydroxylation at their surfaces. This letter explores the relationship between the hydroxylation of oxide surfaces and their resulting wettability. Here we demonstrate that hydroxyls increase the hydrophobicity, or reduce the wettability, of oxide surfaces by reducing the polar component of surface free energy. Using alumina as a model material, increased hydrophobicity with hydroxylation was confirmed experimentally and a correlation between the strength of the hydroxyl-driven hydrophobic response and surface treatment was demonstrated. PMID- 20000329 TI - Photon and electron spins. AB - It is easy to draw an intuitive parallel between the classical free electromagnetic field and its corresponding quantum, the photon-a spin-1 object. The situation with a massive particle such as an electron is less clear, as a real-world analog of the classical field whose quantum is the massive particle is not available. It is concluded that the fermion particle perspective provides the best avenue for an intuitive grasp of the spin of an elementary fermion. PMID- 20000330 TI - Hydrogen bond interactions of the pheophytin electron acceptor and its radical anion in photosystem II as revealed by Fourier transform infrared difference spectroscopy. AB - The primary electron acceptor pheophytin (Pheo(D1)) plays a crucial role in regulation of forward and backward electron transfer in photosystem II (PSII). It is known that some cyanobacteria control the Pheo(D1) potential in high-light acclimation by exchanging the D1 proteins from different copies of the psbA genes. To clarify the mechanism of the potential control of Pheo(D1), we studied the hydrogen bond interactions of Pheo(D1) in the neutral and anionic states using light-induced Fourier transform infrared (FTIR) difference spectroscopy. FTIR difference spectra of Pheo(D1) upon its photoreduction were obtained using three different PSII preparations, PSII core complexes from Thermosynechococcus elongatus possessing PsbA1 as a D1 subunit (PSII-PsbA1), those with PsbA3 (PSII PsbA3), and PSII membranes from spinach. The D1-Gln130 side chain, which is hydrogen bonded to the 13(1)-keto C=O group of Pheo(D1) in PSII-PsbA1, is replaced by Glu in PSII-PsbA3 and spinach PSII. The spectrum of PSII-PsbA1 exhibited 13(1)-keto C=O bands at 1682 and 1605 cm(-1) in neutral Pheo(D1) and its anion, respectively, while the corresponding bands were observed at frequencies lower by 1-3 and 18-19 cm(-1), respectively, in the latter two preparations. This larger frequency shift in Pheo(D1)(-) than Pheo(D1) by the change of the hydrogen bond donor was well reproduced by density functional theory (DFT) calculations for the Pheo models hydrogen bonded with acetamide and acetic acid. The DFT calculations also exhibited a higher redox potential for Pheo reduction in the model with acetic acid than that with acetamide, consistent with previous observations for the D1-Gln130Glu mutant of Synechocystis. It is thus concluded that a stronger hydrogen bond effect on the Pheo(-) anion than the neutral Pheo causes the shift in the redox potential, which is utilized in the photoprotection mechanism of PSII. PMID- 20000331 TI - DNA repair by photolyase: a novel substrate with low background absorption around 265 nm for transient absorption studies in the UV. AB - CPD photolyase enzymatically repairs the major UV-induced lesion in DNA, the cyclobutane pyrimidine dimer (CPD), by photoreversion of the damage reaction. An enzyme-bound reduced flavin (FADH(-)) cofactor functions as photosensitizer. Upon excitation, it transiently transfers an electron to the CPD, triggering scission of the interpyrimidine bonds. After repair completion, the electron returns to the flavin to restore its functional reduced form. A major difficulty for time resolved spectroscopic monitoring of the enzymatic repair reaction is that absorption changes around 265 nm accompanying pyrimidine restoration are obscured by the strong background absorption of the nondimerized bases in DNA. Here we present a novel substrate for CPD photolyase that absorbs only weakly around 265 nm: a modified thymidine 10-mer with a central CPD and all bases, except the one at the 3' end, replaced by 5,6-dihydrothymine which virtually does not absorb around 265 nm. Repair of this substrate by photolyases from Anacystis nidulans and from Escherichia coli was compared with repair of two conventional substrates: a 10-mer of unmodified thymidines containing a central CPD and an acetone-sensitized thymidine 18-mer that contained in average six randomly distributed CPDs per strand. In all cases, the novel substrate was repaired with an efficiency very similar to that of the conventional substrates (quantum yields in the order of 0.5 upon excitation of FADH(-)). Flash-induced transient absorption changes at 267 nm could be recorded on a millisecond time scale with a single subsaturating flash and yielded very similar signals for all three substrates. Because of its low background absorption around 265 nm and the defined structure, the novel substrate is a promising tool for fast and ultrafast transient absorption studies on pyrimidine dimer splitting by CPD photolyase. PMID- 20000332 TI - Molecular structure and chemical and electrochemical reactivity of Co2(dpb)4 and Rh2(dpb)4 (dpb=N,N'-Diphenylbenzamidinate). PMID- 20000333 TI - Differences in the band electronic structures of the tetragonal and orthorhombic phases of the high-temperature superconductor yttrium barium copper oxide, YBa2Cu3O7-y (Tc > 90 K). PMID- 20000334 TI - Diagnostics for the developing world: microfluidic paper-based analytical devices. AB - Microfluidic paper-based analytical devices (microPADs) are a new class of point of-care diagnostic devices that are inexpensive, easy to use, and designed specifically for use in developing countries. (To listen to a podcast about this feature, please go to the Analytical Chemistry multimedia page at pubs.acs.org/page/ancham/audio/index.html.). PMID- 20000335 TI - From silphinenes to penifulvins: a biomimetic approach to penifulvins B and C. AB - The biomimetic total synthesis of penifulvins B and C uses a meta photocycloaddition as a key step which gives rapid access to the fenestrane-type carboskeleton with control of an onring quaternary stereocenter. The route is concise, stereocontrolled, scaleable, and flexibile and requires only one protecting group. PMID- 20000337 TI - Calmodulin-induced conformational and hydrodynamic changes in the catalytic domain of Bordetella pertussis adenylate cyclase toxin. AB - Bordetella pertussis, the causative agent of whooping cough, secretes among various toxins an adenylate cyclase (CyaA) that displays a unique mechanism of cell invasion, which involves a direct translocation of its N-terminal catalytic domain (AC, 400 residues) across the plasma membrane of the eukaryotic targeted cells. Once into the cytosol, AC is activated by endogenous calmodulin and produces toxic amounts of cAMP. The structure of AC in complex with the C terminal part of calmodulin has recently been determined. However, as the structure of the catalytic domain in the absence of calmodulin is still lacking, the molecular basis of AC activation by calmodulin remains largely unknown. To characterize this activation mechanism, we investigated here the biophysical properties of the isolated catalytic domain in solution with or without calmodulin. We found that calmodulin triggered only minor modifications of the protein secondary and tertiary structure but had a pronounced effect on the hydrodynamic properties of AC. Indeed, while the isolated catalytic domain was spherical and hydrated, it underwent a significant elongation as well as compaction and dehydration upon calmodulin interaction. On the basis of these data, we propose a model for the structural transition between the calmodulin free and calmodulin-bound AC. PMID- 20000336 TI - Molecular species of phosphatidylinositol-cycle intermediates in the endoplasmic reticulum and plasma membrane. AB - Phosphatidylinositol (PI) turnover is a process requiring both the plasma and ER membranes. We have determined the distribution of phosphatidic acid (PA) and PI and their acyl chain compositions in these two subcellular membranes using mass spectrometry. We assessed the role of PI cycling in determining the molecular species and quantity of these lipids by comparing the compositions of the two membranes isolated from embryonic fibroblasts obtained from diacylglycerol kinase epsilon (DGKepsilon) knockout (KO) and wild-type (WT) mice. In the KO cells, the conversion of arachidonoyl-rich DAG to PA is blocked by the absence of DGKepsilon, resulting in a reduction in the rate of PI cycling. The acyl chain composition is very similar for PI and PA in the endoplasmic reticulum (ER) versus plasma membrane (PM) and for WT versus KO. However, the acyl chain profile for PI is very different from that for PA. This indicates that DGKepsilon is not facilitating the direct transfer of a specific species of PA between the PM and the ER. Approximately 20% of the PA in the ER membrane has one short acyl chain of 14 or fewer carbons. These species of PA are not converted into PI but may play a role in stabilizing regions of high positive curvature in the ER. There are also PI species in both the ER and PM for which there is no detectable PA precursor, indicating that these species of PI are unlikely to arise via the PI cycle. We find that in the PM of KO cells the levels of PI and of PA are decreased approximately 3-fold in comparison with those in either the PM of WT cells or the ER of KO cells. The PI cycle is slowed in the KO cells; hence, the lipid intermediates of the PI cycle can no longer be interconverted and are depleted from the PI cycle by conversion to other species. There is less of an effect of the depletion in the ER where de novo synthesis of PA occurs in comparison with the PM. PMID- 20000338 TI - Fabrication of monodisperse polymer nanoparticles by membrane emulsification using ordered anodic porous alumina. AB - Uniformly sized droplets of photocurable monomer were obtained through membrane emulsification using highly ordered anodic porous alumina as a membrane. Subsequent polymerization of the monomer generated polymer particles, whose sizes could be controlled by changing the size of the pores in anodic porous alumina. The size distribution was very narrow owing to the uniformity of pore size in the anodic porous alumina used for the emulsification. PMID- 20000339 TI - More fluorous surface modifier makes it less oleophobic: fluorinated siloxane copolymer/PDMS coatings. AB - A copolyacrylate with semifluorinated and polydimethylsiloxane side chains (D5-3) was used as a surface modifier for a condensation-cured PDMS coating. The decyl fluorous group is represented by "D"; "5" is a 5 kDa silicone, and "3" is the mole ratio of fluorous to silicone side chains. Wetting behavior was assessed by dynamic contact angle (DCA) analysis using isopropanol, which differentiates silicone and fluorous wetting behavior. Interestingly, a maximum in surface oleophobicity was found at low D5-3 concentration (0.4 wt %). Higher concentrations result in decreased oleophobicity, as reflected in decreased contact angles. To understand this unexpected observation, dynamic light scattering (DLS) studies were initiated on a model system consisting of hydroxyl terminated PDMS (18 kDa) containing varying amounts of D5-3. DLS revealed D5-3 aggregation to be a function of temperature and concentration. A model is proposed by which D5-3 surface concentration is depleted via phase separation favoring D5-3 aggregation at concentrations >0.4 wt %, that is, the cmc. This model suggests increasing aggregate/micelle concentrations at increased D5-3 concentration. Bulk morphologies studied by scanning electron microscopy (SEM) and atomic force microscopy (AFM) support this model by showing increased aggregate concentrations with increased D5-3 > 0.4 wt %. PMID- 20000340 TI - Developing mixed films of immobilized oligonucleotides and quantum dots for the multiplexed detection of nucleic acid hybridization using a combination of fluorescence resonance energy transfer and direct excitation of fluorescence. AB - Methods have been developed for the simultaneous and selective detection of three target nucleic acid sequences based on mixed films of immobilized quantum dots (QDs) and oligonucleotide probes. CdSe/ZnS QDs were immobilized on optical fibers and conjugated with mixtures of different probe oligonucleotides. Hybridization events were detected using a combination of fluorescence from direct excitation and fluorescence sensitized by resonance energy transfer (FRET). A sandwich assay format was used to associate dye labeled reporter oligonucleotides with probe target hybrids formed at the surface of the optical fiber. One detection channel utilized direct excitation of Pacific Blue and the two other detection channels were based on FRET. In one strategy, green emitting QDs were used as donors with Cy3 and Rhodamine Red-X acceptors. In a second strategy, green and red emitting QDs were coimmobilized and used as donors with Cy3 and Alexa Fluor 647 acceptors, respectively. Selective three-plex detection was demonstrated with both strategies. Several key design criteria that were explored to optimize the relative signal magnitude between channels included: the ratio of probe associated with direct excitation versus probes associated with FRET; the relative amounts of each FRET probe and corresponding spectral overlap; and the photoluminescence ratio between immobilized green and red emitting QDs (where applicable). Careful selection of probe sequences and lengths were important for the discrimination of single nucleotide polymorphisms in one channel without suppressing binding of target in the other two channels. This work provides a basis for the development of multiplexed biosensors that are ensemble compatible and do not require discrete sensor elements, spatial registration, sorting technology, or single molecule spectroscopy. PMID- 20000341 TI - L-cysteine-assisted growth of core-satellite ZnS-Au nanoassemblies with high photocatalytic efficiency. AB - Core-satellite ZnS-Au nanoassemblies, in which each of the ZnS nanospheres was surrounded by a few Au nanoparticles, have been successfully prepared with a facile L-cysteine-assisted hydrothermal approach. The density of Au nanoparticles encircling each ZnS nanosphere can be readily controlled through suitably modulating the concentration of Au added. Because of the difference in band structures between ZnS and Au, a pronounced photoinduced charge separation was observed for the as-synthesized ZnS-Au nanoassemblies. As compared to the relevant commercial products like Au-loaded P-25 TiO(2) and ZnS powders, ZnS-Au nanoassemblies exhibited superior photocatalytic performance, demonstrating their potential as an efficient photocatalyst in relevant redox reactions. Furthermore, the recycling test revealed that core-satellite nanoassemblies of ZnS-Au could be promisingly utilized in the long-term course of photocatalysis. The present study provides a new paradigm for designing the highly efficient semiconductor/metal hybrid photocatalysts that can effectively produce chemical energy from light. PMID- 20000342 TI - Evaporation-induced crystallization of pluronic F127 studied in situ by time resolved infrared spectroscopy. AB - Rapid scan time-resolved infrared spectroscopy has been used to study in situ the crystallization induced by evaporation in an aqueous solution of a triblock copolymer, Pluronic F127. A droplet of the solution was cast on a silicon substrate and the evaporation followed by an infrared microscope in transmission mode. The evaporation rate of water, in the last stage of the process, has been shown to be correlated to the changes in the block copolymer; four different stages can be distinguished. The block copolymer passes from an amorphous micellar state in water to a partially crystallized phase in well-defined stages of the evaporation; the complete change from amorphous to crystalline state of Pluronic F127 is observed only after all water is evaporated. PMID- 20000343 TI - Toward safer processes for C-C biaryl bond construction: catalytic direct C-H arylation and tin-free radical coupling in the synthesis of pyrazolophenanthridines. AB - A series of pyrazolo[1,5-f]phenanthridine derivatives has been efficiently synthesized by a short, straightforward sequence. A tandem amine exchange/heterocyclization of enaminones was successfully applied to the regioselective preparation of 1,5-diarylpyrazole intermediates with structure resemblance to relevant nonsteroidal anti-inflammatory drugs such as celecoxib or tepoxalin. The final key step, cyclization by intramolecular biaryl bond formation, was accomplished by two alternative methodologies: radical coupling and catalytic direct arylation via C-H activation. The scope and limitations of the two methodologies have been explored and their complementariness has been established. In addition, polymer-supported heterogeneous catalysts have been compared with homogeneous analogues. In the radical process, toxic tin derivatives have been avoided in order to employ environmentally safer protocols. PMID- 20000345 TI - Tampering with molecular cohesion in crystals of hexaphenylbenzenes. AB - Hexaphenylbenzene (HPB) and analogous compounds have properties of broad utility in science and technology, including conformationally well-defined molecular structures, high thermal stability, high HOMO-LUMO gaps, little self-association, inefficient packing, and high solubilities. Previous structural studies of HPB and its analogues have revealed persistent involvement of the central aromatic ring in strong C-H...pi interactions. These interactions can be blocked by adding simple ortho alkyl substituents to the peripheral phenyl groups. Comparison of the structures of HPB and a series of ortho-substituted derivatives has shown systematic changes in molecular cohesion and packing, as measured by packing indices, densities, solubilities, temperatures of sublimation, melting points, and ratios of H...H, C...H, and C...C contacts. These results illustrate how crystal engineering can guide the search for improved materials by identifying small but telling molecular alterations that thwart established patterns of association. PMID- 20000344 TI - An LC-IMS-MS platform providing increased dynamic range for high-throughput proteomic studies. AB - A high-throughput approach and platform using 15 min reversed-phase capillary liquid chromatography (RPLC) separations in conjunction with ion mobility spectrometry-mass spectrometry (IMS-MS) measurements was evaluated for the rapid analysis of complex proteomics samples. To test the separation quality of the short LC gradient, a sample was prepared by spiking 20 reference peptides at varying concentrations from 1 ng/mL to 10 microg/mL into a tryptic digest of mouse blood plasma and analyzed with both a LC-Linear Ion Trap Fourier Transform (FT) MS and LC-IMS-TOF MS. The LC-FT MS detected 13 out of the 20 spiked peptides that had concentrations >or=100 ng/mL. In contrast, the drift time selected mass spectra from the LC-IMS-TOF MS analyses yielded identifications for 19 of the 20 peptides with all spiking levels present. The greater dynamic range of the LC-IMS TOF MS system could be attributed to two factors. First, the LC-IMS-TOF MS system enabled drift time separation of the low concentration spiked peptides from the high concentration mouse peptide matrix components, reducing signal interference and background, and allowing species to be resolved that would otherwise be obscured by other components. Second, the automatic gain control (AGC) in the linear ion trap of the hybrid FT MS instrument limits the number of ions that are accumulated to reduce space charge effects and achieve high measurement accuracy, but in turn limits the achievable dynamic range compared to the IMS-TOF instrument. PMID- 20000346 TI - Dihydroxylation of vinyl sulfones: stereoselective synthesis of (+)- and (-) febrifugine and halofuginone. AB - The asymmetric dihydroxylation of amino-functionalized vinyl sulfone 19 has been used for the 3-step preparation of 3-hydroxylpiperidine 24 in 86% enantiomeric excess. This enantiomerically enriched building block was used then to synthesize the naturally occurring antimalarial alkaloid febrifugine 1 and its antiangiogenic analogue, halofuginone 3. PMID- 20000347 TI - Heterostereocomplexation between biodegradable and optically active polyesters as a versatile preparation method for biodegradable materials. AB - The thermal properties and crystallization of biodegradable and optically active poly[(S)-2-hydroxybutyrate] [P(S-2HB)], poly(l-lactide) (PLLA), poly(d-lactide) (PDLA) and their blends were investigated. The results of differential scanning calorimetry, wide-angle X-ray scattering (WAXS), and polarized optical microscopy first indicated heterostereocomplexation between biodegradable and optically active polyesters having different chemical structures and opposite configurations, that is, P(S-2HB) and PDLA. The melting temperature of the heterostereocomplex was higher than those of pure polymers. Such cocrystallization was not observed for P(S-2HB)/PLLA blends having identical configurations. The WAXS profile of P(S-2HB)/PDLA heterostereocomplex was very similar to those of the PLLA/PDLA and P(S-2HB)/P(R-2HB) homostereocomplexes and each crystalline diffraction peak of the heterostereocomplex was located between those of the homostereocomplexes. The present study strongly suggests that heterostereocomplexation will provide a novel versatile method for preparing biodegradable polyester materials with a wide range of physical properties and biodegradability. PMID- 20000348 TI - Palladium-catalyzed alkoxylation of N-methoxybenzamides via direct sp2 C-H bond activation. AB - The palladium-catalyzed ortho-alkoxylation of N-methoxybenzamides has been demonstrated. With the CONHOMe group as a directing group, the aromatic C-H bond can be functionalized efficiently to generate ortho-alkoxylated derivatives in moderate to good yields. PMID- 20000349 TI - Elucidating colorization in the functionalization of hydroxyl-containing polymers using unsaturated anhydrides/acyl chlorides in the presence of triethylamine. PMID- 20000350 TI - Fluorescent conjugated polyfluorene with pendant lactopyranosyl ligands for studies of Ca(2+)-mediated carbohydrate-carbohydrate interaction. AB - A well-defined fluorescent conjugated polyfluorene with pendant lactopyranosyl ligands was easily prepared through Cu(I)-catalyzed azide/alkyne "click" ligation and Suzuki coupling polymerization. As a fluorescent multivalent model system of glycoconjugates, the polymer was first used for studies of metal ion-mediated carbohydrate-carbohydrate interaction based on fluorescence spectroscopy. A significant fluorescence quenching of the lactosyl-bearing polyfluorene was observed upon addition of calcium ion, which is attributed to the polymer aggregation derived from Ca(2+)-mediated complex formation. Dynamic light scattering can also prove Ca(2+)-induced aggregation of the polymer based on determination of the corresponding hydrodynamic diameters. The calcium-mediated lactose-lactose interaction was reversible when treated with EDTA. In control studies, Ca(2+)-induced fluorescence quenching can not be observed for cellobiosyl- or galactosyl-functionalized polymer analogues, which show that specific sugar structures are critical for carbohydrate-metal complex formation. PMID- 20000351 TI - Rhenium- and manganese-catalyzed synthesis of aromatic compounds from 1,3 dicarbonyl compounds and alkynes. AB - We have succeeded in the development of three approaches to the synthesis of aromatic compounds from 1,3-dicarbonyl compounds and alkynes. The first approach is a manganese-catalyzed [2+2+2] cycloaddition between 1,3-dicarbonyl compounds, which have no substituents at the active methylene moiety, and terminal alkynes. This reaction proceeds with high regioselectivity when aryl acetylenes are employed as the alkyne component. The second approach is a rhenium- or manganese catalyzed formal [2+1+2+1] cycloaddition between beta-keto esters and two kinds of alkynes. In this reaction, the aromatic compounds are obtained by the following reaction sequence: (1) insertion of the first alkyne into a carbon carbon single bond of a beta-keto ester, (2) formation of 2-pyranones via intramolecular cyclization with the elimination of ethanol, and (3) Diels-Alder reaction between the formed 2-pyranone and the second alkyne. This reaction provides multisubstituted aromatic compounds in a regioselective manner. The third approach is a rhenium-catalyzed formal [2+2+1+1] cycloaddition reaction from two 1,3-diketones and one alkyne. In this reaction, the aromatic skeleton is constructed from three carbons of the first 1,3-diketone, two carbons of the alkyne, and one carbon of the second 1,3-diketone. PMID- 20000352 TI - Wood plastic composite produced by nonmetals from pulverized waste printed circuit boards. AB - Nonmetals reclaimed from waste printed circuit boards (PCBs) are used to replace wood flour in the production of wood plastic composite (WPC). To evaluate property durability against weather exposure, the effects of accelerated aging process on the properties of WPC are investigated. The results show that filling of nonmetals in WPC improves the flexural strength and tensile strength, and reduces screw withdrawal strength. Before hollow WPC with 15% nonmetals (H-15 WPC) underwent aging process, H-15-WPC had a flexural strength of 25.8 MPa, a tensile strength of 9.8 MPa, a charpy impact strength of 3.4 kJ/m(2), and face/edge screw withdrawal strength of 121/115 N/mm. It is found that flexural strength of H-15-WPC decreases linearly with the increase of accelerated aging cycles, and the effects of aging test on tensile and impact strength of H-15-WPC are minor. For solid WPC, the accelerated aging test decreases screw withdrawal strength slightly. All the results indicate that nonmetals of waste PCBs can be reused as an alternative for wood flour in WPC products rather than resorting to their landfill or combustion. PMID- 20000353 TI - Reactivity of TiO2 rutile and anatase surfaces toward nitroaromatics. AB - The Au-TiO(2) system is a promising catalyst for the synthesis of nitro-aromatic compounds. The adsorption of azobenzene (C(6)H(5)N=NH(5)C(6)) and aniline (C(6)H(5)NH(2)) on two single-crystalline TiO(2) surfaces, anatase (101) and rutile (110), has been investigated with scanning tunneling microscopy (STM), low energy electron diffraction (LEED), and X-ray photoemission spectroscopy (XPS). While azobenzene adsorbs as an intact molecule at low coverages, ordered overlayers of phenyl imide (C(6)H(5)N) form at saturation coverage, indicating that TiO(2) surfaces cleave the N horizontal lineN bond even without the presence of Au. The same superstructures, p(1 x 2) on anatase and c(2 x 2) on rutile, form upon adsorption of aniline, suggesting the formation of the same, or a very similar, reaction intermediate. These results suggest that the main role of the supported Au in catalytic aniline <--> azobenzene conversion is the activation of O(2)/H(2) for de/hydrogenation reactions. PMID- 20000354 TI - [Ir(COD)Cl]2 as a catalyst precursor for the intramolecular hydroamination of unactivated alkenes with primary amines and secondary alkyl- or arylamines: a combined catalytic, mechanistic, and computational investigation. AB - The successful application of [Ir(COD)Cl](2) as a precatalyst for the intramolecular addition of primary as well as secondary alkyl- or arylamines to unactivated olefins at relatively low catalyst loading is reported (25 examples), along with a comprehensive experimental and computational investigation of the reaction mechanism. Catalyst optimization studies examining the cyclization of N benzyl-2,2-diphenylpent-4-en-1-amine (1a) to the corresponding pyrrolidine (2a) revealed that for reactions conducted at 110 degrees C neither the addition of salts (N(n)Bu(4)Cl, LiOTf, AgBF(4), or LiB(C(6)F(5))(4) x 2.5 OEt(2)) nor phosphine coligands served to enhance the catalytic performance of [Ir(COD)Cl](2). In this regard, the rate of intramolecular hydroamination of 1a employing [Ir(COD)Cl](2)/L2 (L2 = 2-(di-t-butylphosphino)biphenyl) catalyst mixtures exhibited an inverse-order dependence on L2 at 65 degrees C, and a zero order rate dependence on L2 at 110 degrees C. However, the use of 5 mol % HNEt(3)Cl as a cocatalyst was required to promote the cyclization of primary aminoalkene substrates. Kinetic analysis of the hydroamination of 1a revealed that the reaction rate displays first order dependence on the concentration of Ir and inverse order dependence with respect to both substrate (1a) and product (2a) concentrations; a primary kinetic isotope effect (k(H)/k(D) = 3.4(3)) was also observed. Eyring and Arrhenius analyses for the cyclization of 1a to 2a afforded DeltaH(double dagger) = 20.9(3) kcal mol(-1), DeltaS(double dagger) = -23.1(8) cal/K x mol, and E(a) = 21.6(3) kcal mol(-1), while a Hammett study of related arylaminoalkene substrates revealed that increased electron density at nitrogen encourages hydroamination (rho = -2.4). Plausible mechanisms involving either activation of the olefin or the amine functionality have been scrutinized computationally. An energetically demanding oxidative addition of the amine N-H bond to the Ir(I) center precludes the latter mechanism and instead activation of the olefin C=C bond prevails, with [Ir(COD)Cl(substrate)] M1 representing the catalytically competent compound. Notably, such an olefin activation mechanism had not previously been documented for Ir-catalyzed alkene hydroamination. The operative mechanistic scenario involves: (1) smooth and reversible nucleophilic attack of the amine unit on the metal-coordinated C=C double bond to afford a zwitterionic intermediate; (2) Ir-C bond protonolysis via stepwise proton transfer from the ammonium unit to the metal and ensuing reductive elimination; and (3) final irreversible regeneration of M1 through associative cycloamine expulsion by new substrate. DFT unveils that reductive elimination involving a highly reactive and thus difficult to observe Ir(III)-hydrido intermediate, and passing through a highly organized transition state structure, is turnover limiting. The assessed effective barrier for cyclohydroamination of a prototypical secondary alkylamine agrees well with empirically determined Eyring parameters. PMID- 20000355 TI - Systematic analysis of public domain compound potency data identifies selective molecular scaffolds across druggable target families. AB - Molecular scaffolds that yield target family-selective compounds are of high interest in pharmaceutical research. There continues to be considerable debate in the field as to whether chemotypes with a priori selectivity for given target families and/or targets exist and how they might be identified. What do currently available data tell us? We present a systematic and comprehensive selectivity centric analysis of public domain target-ligand interactions. More than 200 molecular scaffolds are identified in currently available active compounds that are selective for established target families. A subset of these scaffolds is found to produce compounds with high selectivity for individual targets among closely related ones. These scaffolds are currently underrepresented in approved drugs. PMID- 20000357 TI - "Depolymerization" approach in Mn cluster chemistry: controlled cleavage of a 1D coordination polymer consisting of Mn(8) units in its constituent, discrete Mn(8) complex. AB - The cleavage of a 1D coordination polymer (Mn(II)(2)Mn(III)(6))(n) containing recognizable octanuclear units to obtain the discrete Mn(II)(2)Mn(III)(6) cluster is reported. PMID- 20000356 TI - Facile identification and quantitation of protein phosphorylation via beta elimination and Michael addition with natural abundance and stable isotope labeled thiocholine. AB - Herein, we employ the unique chemical properties of the quaternary amine present in thiocholine (2-mercapto-N,N,N-trimethyl-ethanaminium) in conjunction with alkaline beta-elimination and Michael addition (BEMA) reactions for the specific detection, identification, and quantitation of phosphorylated serine/threonine containing peptides. Through replacement of the phosphate with thiocholine, the negative charge on the phosphopeptide is switched to a quaternary amine containing a permanent positive charge. This strategy resulted in a 100-fold increase in ionization sensitivity during ESI (sub-500 amol/microL detection limit) accompanied by a markedly enhanced production of informative peptidic fragment ions during CID that dramatically increase sequence coverage. Moreover, the definitive localization of phosphorylated residues is greatly facilitated through the generation of diagnostic triads of fragmentation ions resulting from peptide bond cleavage and further neutral loss of either trimethylamine (-59 Da) or thiocholine thiolate (-119 Da) during collision induced dissociation (CID) in tandem mass spectrometry (MS(2) and MS(3)). Synthesis of stable isotope labeled thiocholine enabled the quantitation of protein phosphorylation with high precision by ratiometric comparisons using heavy and light thiocholine. Collectively, this study demonstrates a sensitive and efficient strategy for mapping of phosphopeptides by BEMA using thiocholine through the production of a diagnostic repertoire of unique fragment ions during liquid chromatography-tandem mass spectrometry (LC-MS(2)/MS(3)) analyses, facilitating phosphosite identification and quantitative phosphoproteomics. PMID- 20000358 TI - Novel tripeptide model of nickel superoxide dismutase. AB - Nickel superoxide dismutase (Ni-SOD) catalyzes the disproportionation of superoxide to molecular oxygen and hydrogen peroxide, but the overall reaction mechanism has yet to be determined. Peptide-based models of the 2N:2S nickel coordination sphere of Ni-SOD have provided some insight into the mechanism of this enzyme. Here we show that the coordination sphere of Ni-SOD can be mimicked using the tripeptide asparagine-cysteine-cysteine (NCC). NCC binds nickel with extremely high affinity at physiological pH with 2N:2S geometry, as demonstrated by electronic absorption and circular dichroism (CD) data. Like Ni-SOD, Ni-NCC has mixed amine/amide ligation that favors metal-based oxidation over ligand based oxidation. Electronic absorption, CD, and magnetic CD (MCD) data collected for Ni-NCC are consistent with a diamagnetic Ni(II) center bound in square-planar geometry. Ni-NCC is quasi-reversibly oxidized with a midpoint potential of 0.72(2) V (vs Ag/AgCl) and breaks down superoxide in an enzyme-based assay, supporting its potential use as a model for Ni-SOD chemistry. PMID- 20000360 TI - Enhanced uniformity in arrays of electroless plated spherical gold nanoparticles using tin presensitization. AB - Gold nanoparticle arrays created with electroless gold plating provide a unique means of transforming nanocylinders usually formed in electron beam lithography to spherical nanoparticles. Alone, electroless gold plating is not selective to the substrate and results in the formation of a gold film on all exposed surfaces of an electron beam patterned sample, including the electron resist. Undesired gold plating occurred near patterned features on the substrate surface, which was reduced by increasing post-spin-coat cure time. When the electron resist is removed, some nanocylinders break off with the gold film, leaving partial cylinders or holes in the patterned elements. By presensitizing the substrate surface with tin, gold cylinders may be selectively deposited to the substrate surface without forming a film on the electron resist. Tin presensitized arrays were produced with 47.1 +/- 7.4 nm radius gold nanoparticles with an interparticle distance of 646.0 +/- 12.4 nm. Defects from sheared, missing, and redeposited Au particles associated with the resist removal were minimized, resulting in enhanced size and shape uniformity of pillars and arrays. Hollow particles were eliminated, and relative standard deviation in particle size was reduced by 7.4% on average, while elongation was reduced 12.3% when astigmatism was eliminated. PMID- 20000361 TI - Stability of phosphonic acid self-assembled monolayers on amorphous and single crystalline aluminum oxide surfaces in aqueous solution. AB - The formation of octadecylphosphonic acid (ODPA) self-assembled monolayers (SAMs) and their stability in water has been studied on four distinctly different aluminum oxide surfaces. The aim was to improve the understanding of the state of binding between the phosphonic acid to the oxide surface and how this interaction depends on the structure and termination of the oxide surface. Single crystalline Al(2)O(3)(0001) and Al(2)O(3)(1102) surfaces were compared to amorphous oxide passive films on aluminum and physical vapor deposited (PVD) amorphous aluminum oxide films on gold. The monolayers were adsorbed from ethanol solution, characterized by means of high-resolution in situ atomic force microscopy (AFM), contact angle measurements, polarization modulated infrared reflection absorption spectroscopy (PM-IRRAS), and diffuse reflectance infrared Fourier transform spectroscopy (DRIFTS), and proved to be self-assembled. On Al(2)O(3)(1102) surfaces and amorphous Al(2)O(3) surfaces, the ODPA self-assembled monolayers showed high stability in aqueous environments. However, the adsorbed ODPA monolayers were substituted by the adsorption of interfacial water on the Al(2)O(3)(0001) surface via the intermediate formation of micelles. The different stability of the monolayers in aqueous environments is explained by the variation of interfacial binding states ranging from ionic interactions between phosphonate groups and the positively charged hydrolytated oxide surface to directed coordination bonds between the phosphonate group and Al ions. PMID- 20000359 TI - DNA polymerase fidelity: comparing direct competition of right and wrong dNTP substrates with steady state and pre-steady state kinetics. AB - DNA polymerase fidelity is defined as the ratio of right (R) to wrong (W) nucleotide incorporations when dRTP and dWTP substrates compete at equal concentrations for primer extension at the same site in the polymerase-primer template DNA complex. Typically, R incorporation is favored over W by 10(3)-10(5) fold, even in the absence of 3'-exonuclease proofreading. Straightforward in principle, a direct competition fidelity measurement is difficult to perform in practice because detection of a small amount of W is masked by a large amount of R. As an alternative, enzyme kinetics measurements to evaluate k(cat)/K(m) for R and W in separate reactions are widely used to measure polymerase fidelity indirectly, based on a steady state derivation by Fersht. A systematic comparison between direct competition and kinetics has not been made until now. By separating R and W products using electrophoresis, we have successfully taken accurate fidelity measurements for directly competing R and W dNTP substrates for 9 of the 12 natural base mispairs. We compare our direct competition results with steady state and pre-steady state kinetic measurements of fidelity at the same template site, using the proofreading-deficient mutant of Klenow fragment (KF(-)) DNA polymerase. All the data are in quantitative agreement. PMID- 20000362 TI - Electrostatic interactions affect nanoparticle-mediated toxicity to gram-negative bacterium Pseudomonas aeruginosa PAO1. AB - Nanoscale materials can have cytotoxic effects. Here we present the first combined empirical and theoretical investigation of the influence of electrostatic attraction on nanoparticle cytotoxicity. Modeling electrostatic interactions between cells and 13 nm spheres of zinc oxide nanoparticles provided insight into empirically determined variations of the minimum inhibitory concentrations between four differently charged isogenic strains of Pseudomonas aeruginosa PAO1. We conclude that controlling the electrostatic attraction between nanoparticles and their cellular targets may permit the modulation of nanoparticle cytotoxicity. PMID- 20000363 TI - Toward understanding whether superhydrophobic surfaces can really decrease fluidic friction drag. AB - Superhydrophobic surfaces in nature such as legs of water striders can get an extra supporting force from the deformed water surface they contact, leading to an anticipation of using water-repellent surfaces on ship and even submarine hulls to reduce friction drag. Here, we first fabricate superhydrophobic coatings with microstructures on glass balls by introducing hydrophobic silica nanoparticles into a polyethylene terephthalate (PET) film. Then, the movement of a superhydrophobic ball on and below water surface is investigated and compared with that of a highly hydrophilic normal glass ball. The results reveal that a superhydrophobic ball can fall more slowly under water compared with a normal glass ball, because the dense microbubbles trapped at the solid/water interface around the superhydrophobic ball act not as a reducer, but as an enhancer for the friction drag. In contrast, the faster movement of a superhydrophobic ball on the water surface can be mainly attributed to the great reduction of skin friction owing to the increased area of the solid/atmosphere interface. PMID- 20000364 TI - Scale dependence of omniphobic mesh surfaces. AB - We provide a simple design chart framework to predict the apparent contact angle on a textured surface in terms of the equilibrium contact angle on a chemically identical smooth surface and details of the surface topography. For low surface tension liquids such as methanol (gamma(lv) = 22.7 mN/m) and octane (gamma(lv) = 21.6 mN/m), a solid-liquid-air composite interface on a textured surface is inherently metastable. Thus, on application of a sufficient pressure difference (e.g., an externally applied pressure or a sufficiently large Laplace pressure at small droplet size) the metastable composite interface transitions to a fully wetted interface. A dimensionless robustness factor is used to quantify the breakthrough pressure difference necessary to disrupt a metastable composite interface and to predict a priori the existence of a robust composite interface. The impact of the length scale (radius of the cylindrical features R varying from 18 to 114 microm) and the feature spacing ratio (D(*) = (R + D)/R varying from 2.2 to 5.1, where 2D is the spacing between the cylindrical features) on the robustness is illustrated by performing contact angle measurements on a set of dip-coated wire-mesh surfaces, which provide systematically quantifiable cylindrical texture. The design chart for a given feature size R shows how the two independent design parameters--surface chemistry as revealed in the equilibrium contact angle and texture spacing embodied in the dimensionless spacing ratio (D(*))--can be used to develop surfaces with desirably large values of the apparent contact angle and robustness of the metastable composite interface. Most revealing is the scaling of the robustness with the dimensionless parameter l(cap)/R (where l(cap = (gamma(lv)/rho g)(1/2) is the capillary length), which indicates clearly why, in the consideration of self-similar surfaces, smaller is better for producing omniphobic surfaces that resist wetting by liquids with low surface tension. PMID- 20000365 TI - Variable-focus liquid microlenses with adjustable 3-D curved housings. AB - This paper reports two novel types of variable-focus liquid microlenses with adjustable sensitivity, which consist of a poly(dimethylsiloxane) (PDMS) lens housing with circular sidewall and a liquid with tunable volume stored in it. The type-I lens housing ("in" bridge structure) is molded from water droplet sandwiched between two plates with identical wettability and the type-II lens housing ("out" bridge structure) is molded from solid microsphere. Because of the variable slope of the sidewall, the altitude of the liquid/air interface, which can be regulated by hydrostatic pressure, determines the curvature of the meniscus and hence the focal length (f). The curvature radius of sidewall, which determines the sensitivity, depends on the surface wettability or the radius of solid microsphere, thus, it is adjustable. Both of the two types of lenses have wide dynamic range, and their tuning tendencies are opposite. The experimental demonstrations and theoretical simulations yield good agreement in the key aspects of the optics. PMID- 20000366 TI - Photochemical cycling of iron mediated by dicarboxylates: special effect of malonate. AB - Photochemical redox cycling of iron coupled with oxidation of malonate (Mal) ligand has been investigated under conditions that are representative of atmospheric waters. Malonate exhibited significantly different characteristics from oxalate and other dicarboxylates (or monocarboxylates). Both strong chelating ability with Fe(III) and strong molar absorptivities, but much low efficiency of Fe(II) formation (Phi(Fe(II)) = 0.0022 +/- 0.0009, 300-366 nm) were observed for Fe(III)-Mal complexes (FMCs). Fe(III) speciation calculation indicated that Mal is capable of mediating the proportion between two photoactive species of Fe(III)-OH complexes and FMCs by changing the Mal concentration. Spin trapping electron spin resonance (ESR) experiments proved the formation of both the (.)CH(2)COOH and (.)OH radicals at lower total Mal concentration ([Mal](T)), but only (.)CH(2)COOH at higher concentrations of malonate, providing strong evidence for competition between malonate and OH(-) and subsequent different photoreaction pathways. Once FMCs dominate the Fe(III) speciation, both photoproduction and photocatalyzed oxidation of Fe(II) will be greatly decelerated. There exists an induction period for both formation and decay of Fe(II) until Fe(III)(OH)(2+) species become the prevailing Fe(III) forms over FMCs as Mal ligand is depleted. A quenching mechanism of Mal in the Fe(II) photoproduction is proposed. The present study is meaningful to advance our understanding of iron cycling in acidified carbon-rich atmospheric waters. PMID- 20000367 TI - Triple recognition of B-DNA by a neomycin-Hoechst 33258-pyrene conjugate. AB - Recent developments have indicated that aminoglycoside binding is not limited to RNA, but to nucleic acids that, like RNA, adopt conformations similar to its A form. We further sought to expand the utility of aminoglycoside binding to B-DNA structures by conjugating neomycin, an aminoglycoside antibiotic, with the B-DNA minor groove binding ligand Hoechst 33258. Envisioning a dual groove binding mode, we have extended the potential recognition process to include a third, intercalative moiety. Similar conjugates, which vary in the number of binding moieties but maintain identical linkages to allow direct comparisons to be made, have also been prepared. We report herein novel neomycin- and Hoechst 33258-based conjugates developed in our laboratories for exploring the recognition potential with B-DNA. Spectroscopic studies such as UV melting, differential scanning calorimetry, isothermal fluorescence titrations, and circular dichroism together illustrate the triple recognition of the novel conjugate containing neomycin, Hoechst 33258, and pyrene. This study represents the first example of DNA molecular recognition capable of minor versus major groove recognition in conjunction with intercalation. PMID- 20000368 TI - Dramatic increase of selectivity for heavy lanthanide(III) cations by tuning the flexibility of polydentate chelators. AB - Two novel octadentate ligands have been synthesized by attaching two terminal iminodiacetic groups to either 1,4-diazepane (BCAED) or piperazine (BCAEP) as central scaffold. The introduction of the seven- or six-membered ring into the ligand backbone is expected to modify their overall flexibility and then to affect the stability of the corresponding lanthanide(III) complexes. In this work, thermodynamic stability data are determined for the formation of the complexes of BCAED and BCAEP with La(3+), Nd(3+), Eu(3+), Gd(3+), Ho(3+), and Lu(3+). The ligand BCAED shows a strong binding affinity for Lu(3+) (logK = 20.99), moderate for Gd(3+) (logK = 17.15) and rather weak for La(3+) (logK = 12.77). Thus, the variation of logK across the Ln series assumes the remarkable value of 8.22, the largest so far reported. This points to a predominant role of a suitable size match between the metal ion and the ligand cavity, determined by its structure and flexibility. The ligand BCAEP forms less stable complexes with lanthanide(III) cations although it retains a good selectivity (DeltalogK(La-Lu) = 5.66). The Gd(III) complexes have been investigated in aqueous solution by measuring their relaxivity as a function of pH, at 20 MHz and 25 degrees C. The results can be interpreted very well in terms of the species distribution curves calculated from the thermodynamic data and indicate that in these complexes Gd(3+) is octacoordinated, without any bound water molecule. This coordination geometry is maintained in the solid state as shown by the X-ray crystal structure of [Na(H(2)O)(2)][Gd(BCAED)] where the metal ion is at the center of a bicapped trigonal prism. Finally, the (13)C NMR spectra (9.4 T, 25 degrees C) of the diamagnetic La(3+), Y(3+), and Lu(3+) complexes show that a pronounced stereochemical rigidity is associated with the thermodynamically more stable complexes. PMID- 20000369 TI - Effects of sulfation on the physicochemical and functional properties of psyllium. AB - The sulfation of psyllium was carried out with sulfur trioxide-pyridine in dimethyl formamide. Three sulfated psyllium derivatives, named SP1, SP2, and SP3, were characterized by sulfur content determination, elemental analysis, FT-IR, and surface charge analysis. The sulfated derivatives were also evaluated for their morphological and rheological properties, water uptake capacities, swelling volumes, and in vitro bile acid-binding abilities. The results showed that sulfation reduced the gelling capacity of psyllium and the viscosity of its solution, and significantly increased its bile acid-binding capacity. Sulfation might also increase the water uptake ability of psyllium but might decrease its swelling capacity. The three sulfated psyllium derivatives had in vitro binding capacities against cholic and chenodeoxycholic acids comparable to that of cholestyramine resin on a per same as it is weight basis. The bile acid-binding capacity of SP1 was about 8.4-fold of that observed for the original psyllium preparation under the same assay conditions. The results from this study suggest that sulfation is a possible approach to obtain novel psyllium derivatives with desirable physicochemical, functional, and biological properties for utilization in functional foods or supplemental and pharmaceutical products. PMID- 20000370 TI - Dynamic Monte Carlo simulation for highly efficient polymer blend photovoltaics. AB - We developed a model system for blend polymers with electron-donating and accepting compounds. It is found that the optimal energy conversion efficiency can be achieved when the feature size is around 10 nm. The first reaction method is used to describe the key processes (e.g., the generation, the diffusion, the dissociation at the interface for the excitons, the drift, the injection from the electrodes, and the collection by the electrodes for the charge carries) in the organic solar cell by the dynamic Monte Carlo simulation. Our simulations indicate that a 5% power conversion efficiency (PCE) is reachable with an optimum combination of charge mobility and morphology. The parameters used in this model study correspond to a blend of novel polymers (bis(thienylenevinylene) substituted polythiophene and poly(perylene diimide-alt-dithienothiophene)), which features a broad absorption and a high mobility. The I-V curves are well reproduced by our simulations, and the PCE for the polymer blend can reach up to 2.2%, which is higher than the experimental value (>1%), one of the best available experimental results up to now for the all-polymer solar cells. In addition, the dependency of PCE on the charge mobility and the material structure are also investigated. PMID- 20000371 TI - Molecular cage occupancy of clathrate hydrates at infinite dilution: experimental determination and thermodynamic significance. AB - This study focuses on the cage occupancy of guest molecules in the infinitely dilute state. At the extreme conditions of highly diluted guest concentrations the direct measurements of the cage occupancy ratio representing the competitive inclusion of multiguest species appear to be so difficult because of spectroscopic intensity limitation, but its thermodynamic significance might be considerable due to the fact that the infinite-dilution value of the cage occupancy ratio can provide the valuable thermodynamic information as a very unique and guest-specific parameter. To experimentally identify gaseous guest populations in structure I (sI) and structure II (sII) cages, we used the solid state nuclear magnetic resonance (NMR), gas chromatography, and direct gas measurements. Furthermore, we derived the simple and generalized thermodynamic equation related to cage occupancies at infinite dilution from the van der Waals Platteeuw model. Both experimental and predicted values agree well within the experimental error range. PMID- 20000372 TI - Factors that influence helical preferences for singly charged gas-phase peptide ions: the effects of multiple potential charge-carrying sites. AB - Ion mobility-mass spectrometry is used to investigate the structure(s) of a series of model peptide [M + H](+) ions to better understand how intrinsic properties affect structure in low dielectric environments. The influence of peptide length, amino acid sequence, and composition on gas-phase structure is examined for a series of model peptides that have been previously studied in solution. Collision cross sections for the [M + H](+) ions of Ac-(AAKAA)(n)Y NH(2) (n = 3-6) and Ac-Y(AEAAKA)(n)F-NH(2) (n = 2-5) are reported and correlated with candidate structures generated using molecular modeling techniques. The [M + H](+) ions of the AAKAA peptide series each exhibit a single, dominant ion mobility arrival time distribution (ATD) which correlates to partial helical structures, whereas the [M + H](+) ions of the AEAAKA ion series are composed of ATDs which correlate to charge-solvated globules (i.e., the charge is coordinated or solvated by polar peptide functional groups). These data raise numerous questions concerning intrinsic properties (amino acid sequence and composition as well as charge location) that dictate gas-phase peptide ion structure, which may reflect trends for peptide ion structure in low dielectric environments, such as transmembrane segments. PMID- 20000373 TI - Solvation dynamics of dipolar probes in dipolar room temperature ionic liquids: separation of ion-dipole and dipole-dipole interaction contributions. AB - Recent solvation dynamics experiments with common dipolar solvation probes in imidazolium room temperature ionic liquids (RTIL) have revealed large dynamic Stokes' shifts and biphasic solvation energy relaxations. Because of the dipolar nature of the imidazolium cations, the solute-cation (dipole-dipole) interaction may, in addition to the ion-solute (ion-dipole) interaction, contribute significantly to the observed Stokes' shift and its dynamics. Conventional time resolved measurements, however, cannot separate out these contributions. A simple semimolecular theory is described here which, upon separation of the dipolar part of the solvation energy from the ion-dipole part, allows estimation of these two components in the measured shifts. While the sum-total of these separated out components agree well with experiments, the ion-dipole interaction is found to contribute approximately 60% of the measured shift in each of these dipolar RTILs. In addition, the calculated solvation response functions, as observed in experiments, are characterized by a fast exponential component (approximately 15 20%) with a time constant in the subpicosecond regime and a slow nonexponential component with a time constant in the subnanosecond regime. Interestingly, the present theory finds that the fast component of the solvation response function in imidazolium ionic liquids originates from the rapid orientational relaxation involving the dipolar species in these liquids, whereas the relaxation of the ion dynamic structure factor via ion translation produces the observed slow nonexponential component. In addition, calculations presented here explain why the continuum model based theories of solvation dynamics do not work for these liquids. For alkylphosphonium ionic liquids, the ion-dipole interaction accounts for nearly 75% of the measured shifts. The present theory also explains why the experimentally observed solvation response function in these liquids does not contain any subpicosecond component and decays nonexponentially with only a single relaxation time constant. PMID- 20000374 TI - Metal-complexes as ligands to generate asymmetric homo- and heterodinuclear M(A)(III)M(B)(II) species: a magneto-structural and spectroscopic comparison of imidazole-N versus pyridine-N. AB - Ten hetero- and homodinuclear M(A)(III)M(B)(III) complexes, 1-10, containing the metal centers Fe(III)Zn(II) (1), Fe(III)Cu(II) (2), Fe(III)Ni(II) (3), Fe(III)Fe(II) (4), Fe(III)Mn(II) (5), Cr(III)Ni(II) (6), Cr(III)Zn(II) (7), Ga(III)Ni(II) (8), Co(III)Fe(II) (9), and Mn(III)Mn(II) (10) are described. The tridentate ligation property of the divalent metal complexes tris(1 methylimidazole-2-aldoximato)metal(II) with three facially disposed pendent oxime O-atoms has been utilized to generate the said complexes. Complexes were characterized by various physical methods including MS, IR, UV-vis, Mossbauer and EPR spectroscopy, cyclic voltammetry (CV), variable-temperature (2-290 K) magnetic susceptibility, and X-ray diffraction techniques. Binuclear complexes 1 10 contain three oximato anions as bridging ligands and are isostructural in the sense that they all contain a metal(III) ion, LM(A)(III) (L = 1,4,7-trimethyl 1,4,7-triazacyclononane), in a distorted octahedral environment M(A)(III)N(3)O(3) and a second six-coordinated divalent metal ion, M(B)(II), in a trigonally distorted M(B)(II)N(6) geometry. A comparison of the cyclic voltammograms of the complexes with those of similar systems reveal both ligand-centered and metal centered redox processes. Complexes 2, 3, 5, and 6 display antiferromagnetic exchange coupling of the neighboring metal centers in the order Fe(III)Mn(II) (5) < Fe(III)Ni(II) (3) < Fe(III)Cu(II) (2) whereas Fe(III)Ni(II) (3) > Cr(III)Ni(II) (6). On the contrary, complex 10, containing high-spin Mn(III) and Mn(II) centers, exhibits ferromagnetic coupling yielding a "high-spin" molecule with an S(t) = (9)/(2) ground state. X-band electron paramagnetic resonance (EPR) spectroscopy for 6, Cr(III)Ni(II) and 3, Fe(III)Ni(II) has been used to establish the electronic ground state in great detail and to complement the magnetic susceptibility measurements. Moreover, computational results have been included to compare the sigma-bonding character of the nitrogen lone pair in imidazole containing ligand and the analogous pyridine-containing oxime. PMID- 20000375 TI - Parallel Synthesis of bis-heterocyclic isoxazolylmethyl- and isoxazolinylmethylpyrazoles. AB - The solution-phase parallel synthesis of a 136-member library of isoxazol(in)e CH(2)-pyrazoles is described. X-ray crystallographic structure determination verified the regioselectivities of the N-alkylation and nitrile oxide 1,3-dipolar cycloaddition steps. The construction of these pharmaceutically relevant heterocycles on solid support under microwave irradiation is also demonstrated. The resulting library of drug-like compounds has been added to the National Institutes of Health repository (approximately 10 mg of each with >or=90% purity) for pilot-scale biomedical studies with bioassay data available at the National Center for Biotechnology Information PubChem database. A subset of these compounds has been broadly screened by Dow AgroSciences for herbicidal, fungicidal, and insecticidal activity. PMID- 20000376 TI - Bistripodand amide host for compartmental recognition of multiple oxyanions. AB - A new benzene-based hexasubstituted bistripodal receptor is synthesized and explored as a new generation receptor for multiple anion binding. The solid state crystal structure showed the encapsulation of four nitrate anions in a bistripodand fashion with "ababab" conformation of the receptor, and upon complexation with acetate anions, the receptor adopted less favorable "aaabbb" conformation with two encapsulated acetate ions. PMID- 20000377 TI - Novel ratiometric fluorescent sensor for silver ions. AB - Incorporating azacrown[N,S,O] into furoquinoline fluorophore yields a novel ratiometric fluorescent sensor FQ-crown for silver ions. UV-vis absorption and fluorescence emission investigations indicate that FQ-crown bears the features of a large Stokes shift, about 173 nm, and red-shift up to 50 nm in the emission spectra and high affinity for silver ions (log K = 7.21) in ethanol in comparison with other competitive d(10) metal ions. PMID- 20000379 TI - Development of new stereodiverse diaminocyclitols as inhibitors of influenza virus neuraminidase. AB - A concise and modular approach to synthesize a new type of cyclopentene-based diaminocyclitol library from D-serine and L-serine has been developed, and key steps in this synthesis are an aza-Claisen rearrangement, a ring-closing metathesis, and a Baylis-Hillman reaction. The developed chemistry may offer a unique way to investigate the neuraminidase (NA) mutation by systematically mapping the changes within its binding sites. PMID- 20000378 TI - A short, strong hydrogen bond in the active site of human carbonic anhydrase II. AB - The crystal structure of human carbonic anhydrase II (HCA II) obtained at 0.9 A resolution reveals that a water molecule, termed deep water, Dw, and bound in a hydrophobic pocket of the active site forms a short, strong hydrogen bond with the zinc-bound solvent molecule, a conclusion based on the observed oxygen-oxygen distance of 2.45 A. This water structure has similarities with hydrated hydroxide found in crystals of certain inorganic complexes. The energy required to displace Dw contributes in significant part to the weak binding of CO(2) in the enzyme substrate complex, a weak binding that enhances k(cat) for the conversion of CO(2) into bicarbonate. In addition, this short, strong hydrogen bond is expected to contribute to the low pK(a) of the zinc-bound water and to promote proton transfer in catalysis. PMID- 20000380 TI - The stability of chicken nucleoside triphosphate diphosphohydrolase 8 requires both of its transmembrane domains. AB - Chicken nucleoside triphosphate diphosphohydrolase 8 (NTPDase8) is a cell surface ectonucleotidase with a large extracellular domain (ECD) containing the active site and is anchored to the membrane by two transmembrane domains (TMDs) at the N and C-termini. Unlike other cell surface NTPDases that have been characterized, the chicken NTPDase8 is not susceptible to substrate inactivation or agents that cause membrane perturbation. To determine if the stability of the enzyme is inherent in its ECD, the cDNA construct of the soluble chicken NTPDase8 was expressed and the protein purified. The ATPase activity of the purified soluble chicken NTPDase8 was less than 15% of that of the purified full-length enzyme. Strikingly, in contrast to the membrane-bound enzyme, the activity of the soluble chicken NTPDase8 decreased with time in a temperature-dependent manner as a result of inactivation by ATP, ADP, and P(i). Truncated mutants in which the ECD is anchored by a single TMD at either the N- or the C-terminus by the native chicken NTPDase TMDs or a TMD from a different NTPDase, human NTPDase2, also displayed a nonlinear time course of ATP hydrolysis. While removal of the N- or C terminal TMD affected protein expression differently, the truncated mutants were generally similar to the soluble chicken NTPDase8 with respect to ATP, ADP, and P(i) inactivation. Other biochemical characteristics, e.g., ATPase/ADPase ratios, inhibition by azide, and affinity for ATP, were also altered when one or both of the TMDs were removed from the chicken NTPDase8. These results indicate that (1) both TMDs of the chicken NTPDase8 are required to maintain stability of the enzyme and maximal catalytic activity and (2) the conformations of the ectodomain in the soluble enzyme and the truncated mutants differ from that of the full length chicken NTPDase8. PMID- 20000381 TI - Transport properties of single-file water molecules inside a carbon nanotube biomimicking water channel. AB - The single-file water transport through a biomimic water channel consisting of a (6,6) carbon nanotube (CNT) with different types of external point charges is studied using molecular dynamics simulations. It is demonstrated that, as in the aquaporins, asymmetrically positioned charges cannot generate robust unidirectional water flow in the CNT. Thermal fluctuation in bulk water competes with charge affinity to steer the water transport, resulting in nonmonotonic flow with intermittent reversal of transport direction. The energetic analysis suggests that the water-water interaction, determined by dipole orientation configuration, influences the transport rate significantly. These findings can provide correct biomimic understanding of water transport properties and will benefit the design of efficient functional nanofluidic devices. PMID- 20000383 TI - On the equilibrium of helical nanostructures with ordered mesopores. AB - Helical conformation exists universally at different length scales. We present a new model to explain the energetics of a helical structure with ordered mesopores and successfully predict their equilibrium state. The formation of the helical structure, which is composed of twisted and hexagonally arrayed one-dimensional pore channels, should be understood at the macromorphology level through the competition between surface free energy reduction and torsion strain energy increase. Our model is established by first reverting a helical rod with experimentally defined parameters to a conjectured straight rod without intrinsic pore channel twisting, and then quantitatively calculating the variation of two competitive energies as a function of twist angle in the torsion process starting from the reverted straight rod. Through our model, a free energy curve is achieved, so that the equilibrium state and the helical structural parameters can be predicted, which are in good agreement with experimental results for helical rods synthesized by different surfactant templates. Moreover, our model can be successfully applied to explain the pitch-radius relationships in previous observations. Our achievement provides unique and fundamental understandings for the spontaneous mesoscopic helix formation, which are different from the microscopic helical structures such as DNA chains. PMID- 20000382 TI - Nucleotide excision repair of a DNA interstrand cross-link produces single- and double-strand breaks. AB - The DNA radical resulting from formal abstraction of a hydrogen atom from the thymidine methyl group, 5-(2'-deoxyuridinyl)methyl radical, forms interstrand cross-links with the opposing 2'-deoxyadenosine. This is the first chemically characterized, radical-mediated cross-link between two opposing nucleotides. In addition, cross-linking between opposing bases in the duplex is less common than between those separated by one or two nucleotides. The first step in cross-link repair was investigated using the UvrABC bacterial nucleotide excision repair system. UvrABC incised both strands of the cross-linked DNA, although the strand containing the cross-linked purine was preferred by the enzyme in two different duplexes. The incision sites in one strand were spaced 11-14 nucleotides apart, as is typical for UvrABC incision. The majority of incisions occur at the third phosphate from the 3'-side of the cross-link and eighth or ninth phosphate on the 5'-side. In addition, cleavage was found to occur on both strands, producing double-strand breaks in approximately 25-29% of the incision events. This is the first example of double-strand cleavage during nucleotide excision repair of cross-linked DNA that does not already contain a strand break in the vicinity of the cross-link. PMID- 20000384 TI - The kinetic effect of internal hydrogen bonds on proton-coupled electron transfer from phenols: a theoretical analysis with modeling of experimental data. AB - Proton-coupled electron transfer (PCET) was studied in two biomimetic covalently linked Ru(bpy)(3)-tyrosine complexes with the phenolic proton hydrogen-bonded to an internal carboxylate group. The phenolic group is either a salicylic acid (o hydroxybenzoic acid, SA) or an o-hydroxyphenyl-acetic acid (PA), where the former gives a resonance-assisted hydrogen bond. Transient absorption data allowed direct determination of the rate constant for these intramolecular, bidirectional, and concerted PCET (CEP) reactions, as a function of temperature and H/D isotope. We found, unexpectedly, that the hydrogen bond in SA is in fact weaker than the hydrogen bond in the complex with PA, which forced us to reassess an earlier hypothesis that the proton coupling term for CEP with SA is increased by a stronger hydrogen bond. Consequently, the kinetic data was modeled numerically using a quantum mechanical rate expression. Sufficient experimentally determined observables were available to give robust and well-determined parameter values. This analysis, coupled with DFT/B3LYP and MP2 calculations and MD simulations, gave a detailed insight into the parameters that control the CEP reactions, and the effect of internal hydrogen bonds. We observed that a model with a static proton-tunneling distance is unable to describe the reaction correctly, requiring unrealistic values for the equilibrium proton-tunneling distances. Instead, when promoting vibrations that modulate the proton donor acceptor distance were included, satisfactory fits to the experimental data were obtained, with parameter values that agree with DFT calculations and MD simulations. According to these results, it is in fact the weaker hydrogen bond of SA which increases the proton coupling. The inner reorganization energy of the phenolic groups is a significant factor contributing to the CEP barriers, but this is reduced by the hydrogen bonds to 0.35 and 0.50 eV for the two complexes. The promoting vibrations increase the rate of CEP by over 2 orders of magnitude, and dramatically reduce the kinetic isotope effect from ca. 40 for the static case to a modest value of 2-3. PMID- 20000385 TI - Theoretical study on rotation of pyrrole rings in porphyrin and N-confused porphyrin. AB - Rotation of pyrrole rings in regular porphyrins and N-confused porphyrins is theoretically investigated by DFT calculations. While the inversion of the pyrrole rings in the regular porphyrins requires high activation energies (36.5 49.1 kcal/mol), the inversion of the confused pyrrole rings in the N-confused porphyrins requires much lower activation energies (18.1-24.5 kcal/mol). This marked difference can be explained by the intramolecular hydrogen bondings and aromatic stabilization due to the [18]annulenic substructures, where confusion and NH tautomerism play an important role. In both of the macrocycles, 360 degrees rotation of their pyrrole rings would be difficult possibly due to the small cavity. Alternatively, a reaction pathway for the production of N-fused porphyrin from N-confused porphyrin is obtained, which is consistent with the experimental observation. PMID- 20000386 TI - Theoretical studies on the structures and the aromaticity for condensed cyclobutadienoids series: the combination of Kekule structures. AB - The structures and aromaticity of a series of condensed cyclobutadienoids were investigated by ab initio molecular orbital and density functional methods. A boat-type structure was found to be the most stable for all cyclobutadienoids except butalene, and the structures could also be predicted from a simple combination of asymmetric Kekule structures. There were found to be three types of stable structure for cyclobutadienoids (C(2n)H(4)). In the case of 2n = 6m (m = 1, 2, ...), the structure consists of a succession of six-membered pi-resonance ring units, while for 2n-4 = 6m, the structure is an assembly of six-membered pi resonance ring units with two "double-bonds" in the center or at the ends of the structure. In all other cases, the structures are of symmetric Kekule type. The aromaticity for each ring was obtained on the basis of the index of deviation from aromaticity. Although all compounds examined here (except for butalene) showed anti- or nonaromatic nature in the whole molecule, the six-membered ring units in some molecules had an aromatic nature similar to that of butalene. PMID- 20000387 TI - Bonding elucidation of the three common acids H2SO4, HNO3, and HClO4. AB - The bonding nature of the non-hydroxyl oxygen atoms of the familiar acids sulfuric [(OH)(2)SO(2)], nitric (HONO(2)), and perchloric (HOClO(3)) is explained without using such ad hoc concepts of circular logic like "hypervalency". It is proposed here for the first time and confirmed through the help of coupled cluster RCCSD(T) calculations, that the S(-O)(2), N(-O)(2), and Cl(-O)(3) bonding occurs by transfer of the electron pair densities of the central atoms, S, N, and Cl to the first excited state, (1)D, of the non-hydroxyl O atoms. PMID- 20000388 TI - A density functional theory study of the magnetic exchange coupling in dinuclear manganese(II) inverse crown structures. AB - The magnetic exchange coupling constants between two Mn(II) centers for a set of five inverse crown structures have been investigated by means of a methodology based on broken-symmetry unrestricted density functional theory. These novel and highly unstable compounds present superexchange interactions between two Mn centers, each one with S = 5/2 through anionic "guests" such as oxygen, benzene, or hydrides or through the cationic ring formed by amide ligands and alkali metals (Na, Li). Magnetic exchange couplings calculated at B3LYP/6-31G(d,p) level yield strong antiferromagnetic couplings for compounds linked via an oxygen atom or hydride and very small antiferromagnetic couplings for those linked via a benzene molecule, deprotonated in either 1,4- or 1,3- positions. Analysis of the magnetic orbitals and spin polarization maps provide an understanding of the exchange mechanism between the Mn centers. The dependence of J with respect to 10 different density functional theory potentials employed and the basis set has been analyzed. PMID- 20000389 TI - Comment on "A highly accurate and analytic equation of state for a hard sphere fluid in random porous media". PMID- 20000390 TI - Directed nanoparticle motion on an interfacial free energy gradient. AB - Using total internal reflection fluorescence microscopy (TIRFM), we have observed the directed motion of 20 nm probe particles on specific regions of surfaces that exhibited strong gradients of hydrophobicity. Patterned surfaces were prepared by selective photodegradation (using a contact photomask) of a hydrophobically modified fused silica surface. The lateral distribution of hydrophobicity was characterized in situ using the selective affinity of amphiphilic probes (i.e., hydrophobic interaction microscopy). Probe particles were observed to move unidirectionally from regions of lower to higher to hydrophobicity over distances of approximately 1 microm when the hydrophobicity gradient was greater than d(cos theta)/dx = 0.05 +/- 0.02 microm(-1), where theta is the water contact angle on the bare surface. Only adsorption events were observed on energetically homogeneous surface regions. PMID- 20000391 TI - Comment on "Hyperquenched glassy water and hyperquenched glassy ethanol probed by single molecule spectroscopy". PMID- 20000392 TI - Nanobeads highly loaded with superparamagnetic nanoparticles prepared by emulsification and seeded-emulsion polymerization. AB - Functional superparamagnetic colloids possessing high saturation magnetization are prepared by emulsification of superparamagnetic nanoparticles (SPM NPs) and heterogeneous polymerization. The colloids consist of a core of densely packed NPs encapsulated within a thin polymer shell. The cores are made by emulsifying SPM NPs and toluene into an aqueous surfactant solution, and subsequently condensing the emulsion droplets by removal of the solvent generating clusters of SPM NPs. By tuning the emulsification condition, this approach allows for control over the size of the clusters from approximately 40 to 200 nm. The polymer shells encapsulating the clusters are made by using seeded-emulsion polymerization concepts. Control over the thickness of the shell and the incorporation of functional groups to the colloid is achieved. Characterization by thermogravimetric analysis (TGA) and magnetometry shows that these colloids have 66 wt % of magnetic material and saturation magnetization of 47 emu/g, confirming that this route generates colloids with a high loading of SPM NPs and high saturation magnetizations. PMID- 20000393 TI - Triadic intramolecular charge transfer compound of tetrathiafulvalene exhibiting multicolor solvatochromism. AB - An A-D-A triad compound comprising a tetrathiafulvalene (TTF) moiety and two pyridyl groups, py-TTF-py (1), has been studied in view of intramolecular charge transfer (ICT). The compound shows a sharp and multicolor change in different solvents and at different pH values, exhibiting good solvatochromism. The results of electronic absorption spectroscopy, cyclic voltammetry, and theoretical calculations confirm that there exists predominantly a monoprotonated A-D-A triad that displays a strong ICT effect, which is tunable as a function of pH. The equilibrium of protonation of 1 has been further studied by means of pH titration, and the result confidently supports the conclusion that only one equivalent H(+) combines with py-TTF-py in dilute solution, even when excessive acid is added. However, unlike the state in dilute solution, the crystal structure of the protonated 1 is a diprotonated A-D-A triad, (1.2H(+)).(CF(3)SO(3)(-))(2).H(2)O (2), that has been structurally characterized. PMID- 20000394 TI - Dynamic theory of liquid junction potentials. AB - A Nernst-Planck-Poisson finite difference simulation system is used to model the dynamic evolution of a liquid junction from a nonequilibrium initial condition to a condition of steady potential difference, in a linear semi-infinite space. Liquid junctions of Lingane's type 1 (monophasic, unequal concentration) and type 2 (bi-ionic potential; biphasic, equal concentration) are considered, for the sake of simplicity. Analysis of the results shows consistency with known and novel asymptotic solutions. A comprehensive dynamic theory of the free liquid junction potential is presented, having considered the simulated concentration profiles and electric field in the system. This reveals a dynamically relaxing junction in which a diffuse layer continues to expand. This is advocated as physically realistic and shown to be consistent with a steady state potential difference, which arises after 10-1000 ns for typical aqueous systems, when the expanding diffuse layer has a corresponding size of 10-1000 nm. Hence, Planck's concept [Wied. Ann. 1890, 40, 561-576] that a steady state potential difference exclusively implies a static junction with equal fluxes of all species is shown to be false, for an unconstrained system. PMID- 20000395 TI - Thermodynamics of glass/crystal transformation in Se58Ge(42-x)Pb(x) (9 < or = x < or = 20) glasses. AB - This article reports the thermodynamics of Se(58)Ge(42-x)Pb(x) (9 < or = x < or = 20) glassy alloys determined from the heat of fusion and specific heat capacity measurements. A differential scanning calorimetry method has been employed for the determination of thermodynamic quantities such as entropy, enthalpy, and Gibbs free energy differences between the glassy and crystalline phase of these alloys as a function of temperature. An effort has also been made to determine the stability of these glasses using the data obtained from different thermodynamic quantities. This study reveals that stability of the samples increases with the increase of lead (Pb) content in the glassy alloys. PMID- 20000396 TI - Direct and selective immobilization of proteins by means of an inorganic material binding peptide: discussion on functionalization in the elongation to material binding peptide. AB - Using an artificial peptide library, we have identified a peptide with affinity for ZnO materials that could be used to selectively accumulate ZnO particles on polypropylene-gold plates. In this study, we fused recombinant green fluorescent protein (GFP) with this ZnO-binding peptide (ZnOBP) and then selectively immobilized the fused protein on ZnO particles. We determined an appropriate condition for selective immobilization of recombinant GFP, and the ZnO-binding function of ZnOBP-fused GFP was examined by elongating the ZnOBP tag from a single amino acid to the intact sequence. The fusion of ZnOBP with GFP enabled specific adsorption of GFP on ZnO substrates in an appropriate solution, and thermodynamic studies showed a predominantly enthalpy-dependent electrostatic interaction between ZnOBP and the ZnO surface. The ZnOBP's binding affinity for the ZnO surface increased first in terms of material selectivity and then in terms of high affinity as the GFP-fused peptide was elongated from a single amino acid to intact ZnOBP. We concluded that the enthalpy-dependent interaction between ZnOBP and ZnO was influenced by the presence of not only charged amino acids but also their surrounding residues in the ZnOBP sequence. PMID- 20000397 TI - Vapor pressure measurements on low-volatility terpenoid compounds by the concatenated gas saturation method. AB - The atmospheric oxidation of monoterpenes plays a central role in the formation of secondary organic aerosols (SOAs), which have important effects on the weather and climate. However, models of SOA formation have large uncertainties. One reason for this is that SOA formation depends directly on the vapor pressures of the monoterpene oxidation products, but few vapor pressures have been reported for these compounds. As a result, models of SOA formation have had to rely on estimated values of vapor pressure. To alleviate this problem, we have developed the concatenated gas saturation method, which is a simple, reliable, high throughput method for measuring the vapor pressures of low-volatility compounds. The concatenated gas saturation method represents a significant advance over traditional gas saturation methods. Instead of a single saturator and trap, the concatenated method uses several pairs of saturators and traps linked in series. Consequently, several measurements of vapor pressure can be made simultaneously, which greatly increases the rate of data collection. It also allows for the simultaneous measurement of a control compound, which is important for ensuring data quality. In this paper we demonstrate the use of the concatenated gas saturation method by determination of the vapor pressures of five monoterpene oxidation products and n-tetradecane (the control compound) over the temperature range 283.15-313.15 K. Over this temperature range, the vapor pressures ranged from about 0.5 Pa to about 70 Pa. The standard molar enthalpies of vaporization or sublimation were determined by use of the Clausius-Clapeyron equation. PMID- 20000398 TI - Comparison of earthworm bioaccumulation between readily desorbable and desorption resistant naphthalene: implications for biouptake routes. AB - The bioaccumulation of readily desorbable naphthalene and that of desorption resistant naphthalene in earthworms were compared to examine the effect of desorption resistance on bioavailability of hydrophobic organic contaminants in soil. A series of naphthalene-contaminated soil samples representing different degrees of desorption resistance were prepared using a batch sorption-repetitive desorption approach, and bioaccumulation of these samples was evaluated using earthworms (Eisenia fetida). Soil samples representing high-degree of desorption resistance exhibited considerably lower bioavailability, as indicated by the lower body burden (naphthalene concentration in worm tissue) at a given sorbed phase naphthalene concentration. Moreover, the body burden of the highly desorption-resistant samples exhibited a significantly different pore-water dependency than that of the readily desorbable samples, indicating that different biouptake mechanisms are likely controlling readily desorbable contaminants and desorption-resistant contaminants. We propose that for readily desorbable contaminants, the primary biouptake route is the pore-water uptake, but for desorption-resistant contaminants enhanced uptake from ingested soil particles can also be important. The surfactant-like substances in worm gut fluids and physical stress such as abrasion are the likely causes for the enhanced release of desorption-resistant contaminants in worm guts. The difference in bioavailability between readily desorbable and desorption-resistant contaminants needs to be taken into account in risk assessment practices. PMID- 20000400 TI - Chemical reactions on surface molecules attached to silicon quantum dots. AB - This Article describes research on chemical reactions on molecules attached to the surface of silicon quantum dots that have been performed to produce quantum dots with reactive surface functionalities such as diols and epoxides. Characterization of the surface reactions includes NMR and FT-IR studies, and the quantum dots were characterized by transmission electron microscopy (TEM) and energy dispersive spectroscopy (EDS). Cytotoxicity and cell viability assay conducted on silicon dots capped with polar molecules indicated low toxicity with quantum dots with more reactive functionalities found to be more toxic. The silicon quantum dots photoluminesce and have been used as a blue chromophore for the biological imaging of cells. PMID- 20000401 TI - In-capillary derivatization and stacking electrophoretic analysis of gamma aminobutyric acid and alanine in tea samples to redeem the detection after dilution to decrease matrix interference. AB - An in-capillary derivatization and stacking capillary electrophoresis (CE) technique has been applied to redeem the detection of dilute analytes in the analysis of gamma-aminobutyric acid (GABA) and alanine (Ala) in tea samples. Extracts from samples were diluted to eliminate matrix interference before introduction into the CE system. GABA and Ala in the diluted sample zone were derivatized with o-phthaldialdehyde/2-mercaptoethanol (OPA/2-ME) to form fluorescence-labeled products in the stacking process, and the labeled derivatives were then enriched by online stacking. Optimal conditions for the stacking, such as the concentration of the background buffer solution, the matrix of the sample zone (sample solution), and the volume of the sample injection, were investigated and then applied to real sample analysis. Under optimum conditions, the detections were linear in the range of 5.0 nM-2.5 microM with the square of correlation coefficients (R2) of 0.9995 and 0.9992 for GABA and Ala, respectively. Detection limits were found to be 0.7 and 0.8 nM for GABA and Ala, respectively. Tea samples were analyzed with recoveries between 92.33 and 97.87% and between 94.36 and 96.46% for GABA and Ala, respectively. This method is a rapid, convenient, and sensitive process for determining GABA and Ala in complicated matrix samples such as tea samples. PMID- 20000402 TI - Anthocyanin and flavonol variation in bog bilberries (Vaccinium uliginosum L.) in Finland. AB - The flavonoids, anthocyanins and flavonols, in bog bilberries (Vaccinium uliginosum L.) were studied from 15 populations in Finland on a south-north axis of approximately 1000 km. Four anthocyanidin xylosides and 14 flavonol glycosides were tentatively identified by means of HPLC-ESI-MS. Twenty-five major flavonoids were quantified by HPLC-DAD. The averages (+/-standard deviation) in the contents of anthocyanins and flavonols were 1425+/-398 and 1133+/-290 mg/100 g of dry weight, respectively. The most abundant anthocyanidin was malvidin, followed by delphinidin, petunidin, cyanidin, and peonidin. Quercetin was the major flavonol, followed by myricetin, laricitrin, syringetin, and isorhamnetin. Anthocyanins were mostly glucosides, whereas flavonols were mainly conjugated to galactose. The anthocyanin content in the berries from the south was the lowest. The delphinidin content was the highest but the proportion of malvidin the lowest in the north. The total flavonol content and the level of myricetin and quercetin were the highest in the north. PMID- 20000403 TI - Quantum chemical molecular dynamics simulations of dynamic fullerene self assembly in benzene combustion. PMID- 20000404 TI - Chirality effects in atomic vacancy-limited transport in metallic carbon nanotubes. AB - We use first principles density functional theory combined with nonequilibrium Green's function technique to investigate the electronic and transport properties of metallic armchair and zigzag carbon nanotubes (CNTs) with different kinds of multivacancy defects. While the existence of a small band gap in pristine zigzag (12,0) CNTs lowers its conductance compared to pristine armchair (7,7) CNTs, transport properties in the presence of multi (hexa)-vacancy are superior in the former nanostructure, that is more sensitive to defect size and topology than the latter. In addition, in the zigzag structures hexavacancy nanotubes have higher conductance than divancancy nanotubes, which is due to the presence of midgap states that reduce the transmission gap and enhance the conductance. PMID- 20000405 TI - Cyclometalated Ru(II) complexes with improved octahedral geometry: synthesis and photophysical properties. AB - Cyclometalated bis-tridentate ruthenium(II) complexes incorporating 2,6 diquinolin-8-ylpyridine ligands and exhibiting broad visible absorptions are described. A [Ru(N(wedge)N(wedge)N)(N(wedge)C(wedge)N)](+) complex based only on ligands with expanded bite angles has a metal-to-ligand charge-transfer excited state lifetime of 16 ns, which is attributed to a strong ligand field and therefore reduced deactivation via metal-centered states. PMID- 20000406 TI - Production of a novel cold-active lipase from Pichia lynferdii Y-7723. AB - Lipase (triacylglycerol acylhydrolases, E.C. 3.1.1.3) is one of the most important enzymes applied to a broad range of industrial application fields. Especially, lipases with abnormal functionality such as thermostability and alkaline, acidic, and cold activities gain special attention because of their applicability in the restricted reaction conditions. In this study, 16 yeast strains prescreened for lipase induction were investigated for their actual lipase production, and we found a novel cold-active lipase produced from Pichia lynferdii Y-7723. The activity of lipase Y-7723 was retained by 74 and 70% at 20 and 10 degrees C, respectively, as compared to the maximum value at 35 degrees C. On the basis of an optimization study, the optimal lipase productivity was obtained at 96 h of incubation with 3% oil substrate in a medium composed of sucrose as a carbon source at pH 7.0. Among carbon sources tested, sucrose showed almost twice as high of a lipase production (184%) as the control, while the cell growth was similar (105%). Yeast extract and ammonium salts were effective as individual nitrogen sources for lipase production. This study demonstrated that the cold activity of lipase Y-7723 at 10 degrees C was highest among the cold active lipases reported so far. PMID- 20000407 TI - Capillary torque caused by a liquid droplet sandwiched between two plates. AB - Capillary force makes a liquid droplet's surface have the minimum area. If the droplet is sandwiched between two plates, it exerts capillary force on the plates. The magnitude of the force depends on the shape of the sandwiched droplet, which is in turn determined by the shape of the plates and the volume of the liquid. The liquid's shape, however, is hard to determine analytically. In this paper, the torque caused by a droplet sandwiched between two noncircular plates is experimentally and theoretically analyzed. We patterned a magnetic material on the surface of the plates and used it to apply a magnetic force to the plates. The torque on the plates was measured. The torque caused by capillary force was calculated by observing the equilibrium between the capillary force and magnetic force. We obtained approximate theoretical solutions for the liquid's shape and torque and verified that they were in accordance with the experimental results. The experimental and theoretical results presented in this paper are useful for designing microdevices or self-assemblies actuated by capillary force. PMID- 20000408 TI - Synthesis and antiviral bioactivities of 2-cyano-3-substituted-amino(phenyl) methylphosphonylacrylates (acrylamides) containing alkoxyethyl moieties. AB - An efficient reaction under microwave irradiation has been developed for the synthesis of a series of novel 2-cyano-3-substituted-amino(phenyl) methylphosphonylacrylates (acrylamides) II. The products obtained in shorter reaction time with moderate yields are fully characterized by elemental analysis, IR, (1)H, (13)C, and (31)P NMR spectral data. The role of introducing various substituents and the effect of incorporating alpha-aminophosphonates with an alkoxyethyl moiety into the parent cyanoacrylate (acrylamide) structure are investigated. Among the studied compounds, both II-17 and II-24 displayed good in vivo curative, protection, and inactivation effects, which were comparable to those of the commercial reference ningnanmycin (inhibitory rates of 58.8, 60.2, 78.9% and 60.0, 58.9, 85.5%, respectively, at 500 mg/L against TMV). To the best of the authors' knowledge, this is the first report on the synthesis and antiviral activity of the title compounds II. PMID- 20000409 TI - Streaming potential effect on the drainage of thin liquid films stabilized by ionic surfactants. AB - Dynamic effects originating from the electric double layers (EDL) are studied in thin liquid films (TLF) containing ionic and nonionic surfactants. To account for such effects, the EDL are to be incorporated into the differential equations describing the TLF drainage. Numerical simulations in the literature have shown that foam films containing ionic surfactants can drain at a slower rate than that predicted by the Reynolds equation (V(Re)) which postulates rigid planar film surfaces. However, the physical reason of the trend has remained unclarified, and the numerical results have not been validated by any experimental data. In the present study, experiments on the drainage of planar foam films were conducted with the anionic surfactant sodium dodecylsulfate (SDS) in the presence of additional electrolyte (0.02 M NaCl) and with the cationic tetrapentylammonium bromide (TPAB). The obtained results are in accord with the numerical simulations from the literature (V/V(Re) < 1). Such behavior was observed already in our preceding experiments on planar TLF with SDS without added electrolyte. These results were compared to the data of the experiments with TLF containing nonionic surfactant, and differences in the drainage pattern between ionics and nonionics were established. A new theoretical model was developed to account for the dynamic effects arising from EDL. According to the present model, the liquid outflow drags the bulk charges of EDL toward the film border, thus generating streaming potential (as in capillary tubes), which in turn brings the charges back toward the center to maintain the state of zero total electrical current. This creates reverse convection of the liquid near the surfaces, resulting in a velocity of film drainage smaller than V(Re). The present theory predicts kinetic dependence closer to the experiment than the Reynolds equation. The limitations of this new model are specified: it is valid for high ionic strength or low value of the surface potential. PMID- 20000410 TI - Design and synthesis of novel insecticides based on the serotonergic ligand 1-[(4 aminophenyl)ethyl]-4-[3-(trifluoromethyl)phenyl]piperazine (PAPP). AB - 1-[(4-Aminophenyl)ethyl]-4-[3-(trifluoromethyl)phenyl]piperazine (PAPP) is a 5 HT(1A) agonist and was reported to display high affinity for serotonin (5-HT) receptor from the parasitic nematode Haemonchus contortus . The present investigation explored the possibility of using PAPP as a lead compound of new insecticides with novel mode of action. On the basis of the PAPP scaffold, a series of 1-arylmethyl-4-[(trifluoromethyl)pyridin-2-yl]piperazine derivatives were designed, synthesized, and evaluated for biological activities against the armyworm Pseudaletia separata (Walker). Bioassays showed that most of the target compounds displayed certain growth-inhibiting activities or larvicidal activities against armyworm. The quantitative structure-activity relationship (QSAR) for growth-inhibiting activities was also analyzed and established. PMID- 20000411 TI - Effect of storage on the content of polyphenols of minimally processed skin-on apple wedges from ten cultivars and two growing seasons. AB - In this study, the polyphenolic composition of skin-on apple wedges from ten cultivars was examined during chill storage and over two growing seasons. Individual polyphenol compounds were measured using HPLC resulting in the total polyphenolic index (TPI). Total phenolic content (TPC) was quantified using the Folin-Ciocalteu assay. Chilled storage had a significant effect (P < 0.001) on the polyphenol composition of all ten cultivars grown in 2007 and 2008. Total phenolic indices (sum of individual polyphenols) and TPCs of nine of the ten cultivars significantly decreased (P < 0.001) after 5 days of storage at 2-4 degrees C. These indices increased in case of Shampion apples over the same storage period. Changes in the most abundant compounds (-)-epicatechin, procyanidins and chlorogenic acid were largely responsible for changes in overall TPI. Percentage loss was higher for compounds such as phloridzin with a degradation of up to 100%. Irrespective of the different starting level of specific polyphenols in each year; storage resulted in a similar percentage loss/gain for each cultivar. PMID- 20000412 TI - Dynamic NMR study of ethene exchange in cationic CNN-type platinum(II) complexes. AB - Cationic ethylene platinum(II) complexes of the type [Pt(CNN)(C(2)H(4))](+), containing a methyl fragment and different diimines (NN), or terdentate (kappaC kappa(2)NN') anionic ligands, were synthesized and fully characterized both as solids and in solution [NN = 2,2'-dipyridylamine, 1; 2,2'-dipyridylsulfide, 2; 1,10-phenanthroline, 3; 4,7-diphenyl-1,10-phenanthroline, 4; 3,4,7,8-tetramethyl 1,10-phenanthroline, 5; 2,2'-bipyridine, 6; HC-NN = 6-tert-butyl-2,2'-bipyridine, 7; 6-neo-pentyl-2,2'-bipyridine, 8; 6-phenyl-2,2'-bipyridine, 9; 6-(alpha methyl)benzyl-2,2'-bipyridine, 10; 6-(alpha-ethyl)benzyl-2,2'-bipyridine, 11; 6 (alpha,alpha-dimethyl)benzyl-2,2'-bipyridine, 12]. Crystals suitable for X-ray analysis of complexes 5 and 7 were obtained. Ethene exchange at the cyclometalated platinum(II) complexes 7, 8, and 10-12 was studied by (1)H NMR line-broadening experiments in chloroform-d, as a function of both temperature and olefin concentrations. For the other prepared complexes the process was too fast to be monitored on the NMR time scale even at the lowest temperature. The ethylene exchange rates show a linear dependence on the concentration of the free ligand, with a negligible k(1) term indicating that either a solvolytic or a dissociative pathway to the products is absent or negligible. The values of the second-order rate constants k(exc), as obtained by linear regression analysis of the experimental data at 298 K, are in a range of ca. 10(4)-10(5) s(-1) m(-1). The activation entropies are negative, ranging between -129 and -112 J K(-1) mol( 1), as expected for associative processes. The activation process is largely entropy controlled: the TDeltaS()() contribution to the free energy of activation is extremely large, amounting to more than 80% for all complexes, with a smaller enthalpy contribution. All the experimental findings evidence that the mechanism takes place via an associative attack by the entering olefin, through a well ordered, stable pentacoordinated transition state with the two ethene molecules on the trigonal plane. The reactivity of [Pt(CNN)(C(2)H(4))](+) complexes is strongly dependent on the choice of coordinated 6-substituted-2,2'-bipyridines, especially when the terdentate anionic fragment is capable of generating steric crowding and congestion on the coordination plane. PMID- 20000413 TI - Synthesis and antiviral activities of phenanthroindolizidine alkaloids and their derivatives. AB - Racemic phenanthroindolizidine alkaloids tylophorine, antofine, and deoxytylophorinine, and optically pure alkaloids S-(+)-tylophorine and R-(-) tylophorine were synthesized and evaluated for their antiviral activities against tobacco mosaic virus (TMV). Further salinization modifications based on tylophorine increased stability and water solubility and improved the antiviral activity in application. The bioassay results showed that most of these synthesized compounds showed higher antiviral activity against TMV in vitro and in vivo than commercial Ningnanmycin. Especially, tylophorine salt derivatives 10, 11, 13, 17, and 22 emerged as potential inhibitors of plant virus. These findings demonstrate that these phenanthroindolizidine alkaloids and their salt derivatives represent a new template for antiviral studies and could be considered for novel therapy against plant virus infection. PMID- 20000414 TI - Synthesis, crystal structure, and insecticidal activities of highly congested hexahydroimidazo[1,2-a]pyridine derivatives: effect of conformation on activities. AB - A series of hexahydroimidazo[1,2-a]pyridine derivatives were designed and synthesized through aza-Diels-Alder reactions and evaluated for insecticidal activities. Compounds 6a-d with endo-conformation were endowed with excellent insecticidal activities against cowpea aphid ( Aphis craccivora ) and armyworm ( Pseudaletia separata Walker), whereas exo-compounds 7a-d showed only low activities against cowpea aphid. The difference in activities between the endo- and exo-conformations indicated that conformation was the determinant of life or death of the insects for these compounds. PMID- 20000415 TI - Screening rules for leads of fungicides, herbicides, and insecticides. AB - To increase efficiency of finding leads in pesticide design, reasonable screening rules for leads of fungicide, herbicide, and insecticide, respectively, are desired. Previous works showed that "Rule 5" of Lipinski is not a suitable screening rule for leads of pesticide and proposed rules for leads of fungicide, insecticide, and herbicide, which were combined by logarithmic ratio of octanol water partition coefficient (log P), number of hydrogen bond donors, molecular weight, number of hydrogen bond acceptors, polar surface area, carcinogenic toxicity, and mutagenic toxicity. Herein, three sets of screening rules for leads of fungicide, insecticide, and herbicide, respectively, are presented. Each set of screening rules involves seven descriptors, which were selected by Kolmogorov Smirnov test, ANOVA, Kruskal-Wallis test, and Pearson product-moment correlation, from more than 450 descriptors calculated by Codessa. Their accuracies are about 82, 83, and 89%, respectively. PMID- 20000416 TI - Feasibility study for the fractionation of the major human immunoglobulin G subclasses using hydrophobic interaction membrane chromatography. AB - Human immunoglobulin G (IgG) consists of four subclasses, each having specific biological functions and physical properties. Fractionation of these subclasses is challenging, both at analytical and preparative scales. This paper examines the feasibility of separating the major IgG subclasses, i.e., IgG1 and IgG2, by hydrophobic interaction membrane chromatography using environment-responsive membranes. These were resolvable as separate peaks at appropriate eluting conditions. This method could potentially be developed further into a rapid and robust IgG subclass profiling technique, suitable for diagnostic applications. This method could also be used to produce specific subclass enriched IgG. PMID- 20000417 TI - Study of inhibitory effects and action mechanism of the novel fungicide pyrimorph against Phytophthora capsici. AB - The antifungal activity of the novel fungicide pyrimorph, (E)-3-[(2 chloropyridine-4-y1)-3-(4-tert-butylpheny1)acryloyl]morpholin, against Phytophthora capsici was investigated in vitro. Pyrimorph inhibited different stages in the life cycle of P. capsici including mycelial growth, sporangium production, zoospore release, and cystospore germination with EC(50) values of 1.84, 0.17, 4.92, and 0.09 microg mL(-1), respectively. The effect of pyrimorph on mycelial growth was reduced by the addition of different concentrations of ATP, which suggested that the action mechanism of pyrimorph was connected with impairment of the energy generation system. Meanwhile, pyrimorph exhibited certain inhibition on metabolic approaches of Embden-Meyerhof-Parnas (EMP), tricarboxylic acid cycle (TCA), and hexosemonophosphate (HMP) by measuring the oxygen consumption of pyrimorph combining with three representative inhibitors to the metabolic approaches. The results indicated that pyrimorph could inhibit the approach of HMP significantly. Morphological and ultrastructural studies showed that pyrimorph caused excessive septation and swelling of hyphae, distortion and disruption of most vacuoles, thickening and development a multilayer cell wall, and accumulation of dense bodies. These results suggested pyrimorph exhibited multiple modes of action including impairment of the energy generation system and effect on cell wall biosynthesis directly or indirectly. PMID- 20000418 TI - Application of the dipeptidyl peptidase IV (DPPIV/CD26) based prodrug approach to different amine-containing drugs. AB - Here we explore the applicability of the dipeptidyl peptidase IV (DPPIV/CD26) based prodrug approach to a variety of amine-containing drugs. Efficient procedures have been developed for the synthesis of dipeptide and tetrapeptide amide prodrugs including N-acylation protocols of the exocyclic amino function of cytidine and adenosine nucleosides. Our studies demonstrated that XaaPro dipeptides linked to a free amino group present on an aromatic ring or on a sugar entity are prodrugs that efficiently release the parent drug upon conversion by purified DPPIV/CD26 as well as soluble DPPIV/CD26 in bovine and human serum. Vildagliptin, a specific inhibitor of DPPIV/CD26, was able to completely block the hydrolysis of the prodrugs in the presence of purified CD26 but also in human and bovine serum. When the amino group is present on a pyrimidine or purine ring, the dipeptide derivatives are chemically unstable, whereas the tetrapeptide derivatives (i.e., ValProValPro or ValAlaValPro) were much more stable in solution and efficiently converted to the parent drug by the action of DPPIV/CD26. This DPPIV/CD26-directed prodrug technology can be useful to increase solubility of the parent drug molecules and/or to allow better formulation properties. PMID- 20000419 TI - Recent advances in the discovery of competitive protein tyrosine phosphatase 1B inhibitors for the treatment of diabetes, obesity, and cancer. PMID- 20000420 TI - Vesicle budding induced by a pore-forming peptide. AB - We describe, in a system whose uniqueness is that the presence of pores allows the volume to vary as budding proceeds, how phase separation on the surface of spheres extrudes material in the process called "budding". The system is giant phospholipid vesicles (GUVs) containing phase-separated regions of DOPC (soft, liquid) and DPPC (stiff, gel), with cholesterol and without it. Budding is triggered by adding the cationic pore-forming peptide, melittin. Without cholesterol, fluorescence experiments show that melittin selectively binds to the liquid domains, inducing them to form mainly exocytotic monodisperse smaller vesicle buds of this same material, causing the parent GUV to shrink. The effect of cholesterol is to produce just a few large buds following domain coalescence, rather than numerous smaller monodisperse ones. Line tension is experimentally shown to be essential for budding in this multicomponent membrane. PMID- 20000421 TI - Anisotropic etching of silver nanoparticles for plasmonic structures capable of single-particle SERS. AB - The understanding of the localized surface plasmons (LSPs) that occur at the geometrically bounded surface of metal nanoparticles continues to advance as new and more complex nanostructures are found. It has been shown that the oscillation of electrons at the metal dielectric interface is strongly dependent on the size, symmetry, and proximity of nanoparticles. Here, we present a new method to chemically control the shape of silver nanocrystals by using a highly anisotropic etching process. Tuning of the etchant strength and reaction conditions allows the preparation of new nanoparticle shapes in high yield and purity, which cannot be synthesized with conventional nanocrystal growth methods. The etching process produces intraparticle gaps, which introduce modified plasmonic characteristics and significant scattering intensity in the near-infrared. These new silver particles serve as excellent substrates for wavelength-tunable, single-particle surface enhanced Raman spectroscopy (spSERS). PMID- 20000422 TI - Isolation and identification of novel macrocyclic lactones from Streptomyces avermitilis NEAU1069 with acaricidal and nematocidal activity. AB - Bioactivity-guided fractionation of Streptomyces avermitilis NEAU1069 fermentation broth was used to isolate and determine the chemical identity of bioactive constituents with acaricidal and nemotocidal activity. The structures of novel compounds 1 and 2 were determined on the basis of spectroscopic analysis, including 1D and 2D NMR as well as HRESI-MS, ESI-MS of spectrometry analysis, UV and IR spectroscopic analyses, and comparison with data from the literature. The acaricidal activities of the isolated compounds against adult mites and mite eggs were evaluated by mortality and unhatched eggs. The nematocidal activity of the isolated compounds against Caenorhabditis elegans was calculated according to the immobilized rates against the total number of tested nematodes. The results indicated that compounds 1 and 2 exhibited potent acaricidal activity against adult mites, with a mortality of >90% at a concentration of 30 microg/mL. However, compounds 1 and 2 showed only weak acaricidal activity against mite eggs, with unhatched mite egg rates of <60% at a concentration of 100 microg/mL. Compound 2, a hydroxylated derivative at C-23 of 1, possessed a high nematocidal activity against C. elegans, with an immobility of >90% at a concentration of 10 microg/mL. These results demonstrate that compounds 1 and 2, especially compound 2, have potential as pesticides with acaricidal and nematocidal activity. PMID- 20000423 TI - Superhydrophobic properties of nonaligned boron nitride nanotube films. AB - Superhydrophobicity is highly desirable for numerous applications. Here, we report that a semierect but nonaligned boron nitride nanotube (BNNT) film showed superhydrophobicity with contact angle above 170 degrees and a small contact angle hysteresis. This superhydrophobicity was stable over a large range of drop sizes, and the measured critical transition pressure was about 10 kPa. However, the prostrate BNNT films only showed hydrophobicity. The drop retraction behavior during evaporation, the pressure effect on contact angle, the critical transition pressure, the drop impact behavior, and the self-cleaning efficiency between these two kinds of films were systematically investigated and compared. PMID- 20000424 TI - Antibacterial inorganic-organic hybrid coatings on stainless steel via consecutive surface-initiated atom transfer radical polymerization for biocorrosion prevention. AB - To enhance the corrosion resistance of stainless steel (SS) and to impart its surface with antibacterial functionality for inhibiting biofilm formation and biocorrosion, well-defined inorganic-organic hybrid coatings, consisting of a polysilsesquioxane inner layer and quaternized poly(2-(dimethyamino)ethyl methacrylate) (P(DMAEMA)) outer blocks, were prepared via successive surface initiated atom transfer radical polymerization (ATRP) of 3 (trimethoxysilyl)propyl methacrylate (TMSPMA) and 2-(dimethylamino)ethyl methacrylate (DMAEMA). The cross-linked P(TMASPMA), or polysilsesquioxane, inner layer provided a durable and resistant coating to electrolytes. The pendant tertiary amino groups of the P(DMAEMA) outer block were quaternized with alkyl halide to produce a high concentration of quaternary ammonium groups with biocidal functionality. The so-synthesized inorganic-organic hybrid coatings on the SS substrates exhibited good anticorrosion and antibacterial effects and inhibited biocorrosion induced by sulfate-reducing bacteria (SRB) in seawater media, as revealed by antibacterial assay and electrochemical analyses, and they are potentially useful to steel-based equipment under harsh industrial and marine environments. PMID- 20000425 TI - Effective viscosity of a concentrated suspension of uncharged spherical soft particles. AB - We present a theory of the effective viscosity eta(s) of a concentrated suspension of uncharged polymer-coated spherical particles, which are termed uncharged spherical soft particles, in a liquid of viscosity eta on the basis of a cell model. These particles consist of the uncharged particle core of radius a covered with an uncharged polymer layer of thickness d (a polymer-coated particle thus has an inner radius a and an outer radius b = a + d). We assume that polymer segments are regarded as resistance centers, exerting frictional forces--gamma u on the liquid flowing in the polymer layer, where u is the liquid velocity and gamma is the frictional coefficient. We derive an analytic expression for the effective viscosity eta(s) of the suspension, which depends on the radii a and b, and volume fraction phi of the spheres and a parameter lambda = (gamma/eta)(1/2). The obtained expression for eta(s) exhibits the correct limiting behaviors. That is, as phi --> 0, the obtained expression for eta(s) becomes that for a dilute suspension of uncharged soft spheres (Ohshima, Langmuir 2008, 24, 6453). As a --> 0, the obtained viscosity expression becomes that for the case of a concentrated suspension of uncharged porous spheres (Ohshima, Colloids Surf. A: Physicochem. Eng. Aspects 2009, 347, 33). In the further limit of a --> 0 and phi --> 0, the obtained viscosity expression becomes the viscosity expression derived for the case of a dilute suspension of uncharged porous spheres by Natraj and Chen (J. Colloid Interface Sci. 2002, 251, 200). As lambda --> infinity or a --> b, the obtained viscosity expression tends to Simha's result (J. Appl. Phys. 1952, 23, 1020) for a concentrated suspension of uncharged rigid spheres of radius b, while as lambda --> 0, it becomes that for uncharged rigid spheres of radius a. PMID- 20000426 TI - Autophilic effect: wetting of hydrophobic surfaces by surfactant solutions. AB - This paper resolves questions in the literature regarding the autophilic effect (i.e., movement of surfactant past the advancing contact line-leading to an increase in drop radius beyond that due to the advance) and its importance to quasi-static sessile drop wetting. Various systems (SDS, HTAB, and MEGA 10 surfactant solutions at three concentrations each and pure water and ethylene glycol on hydrophobic Teflon and OTS-coated silicon) are probed to determine the existence, time constant, and magnitude of the autophilic effect, using quasi static advancing and receding sessile drops. From spreading results and advancing contact angle measurements, it is inferred that the autophilic effect does not occur for our systems (in contradiction of some literature) for the following reasons. First, no relation exists between the time constant for spreading and surfactant concentration, meaning the spreading seen is likely inertial in cause and not due to surfactants. Second, advancing contact angle decreases between tests on clean surfaces and those pre-exposed to surfactant, ruling out the possibility that the autophilic effect is faster than the advance. Third, spreading is seen after the end of the advance over both clean and pre-exposed surfaces, ruling out the possibility that the autophilic effect is slower than the advance. Finally, the pure liquids spread in a similar fashion to surfactant solutions on Teflon and similar contact angle measurements are seen for surfactant solutions and pure liquids of similar surface tension. PMID- 20000427 TI - Phase behavior of an extended surfactant in water and a detailed characterization of the dilute and semidilute phases. AB - The formation of microemulsions with triglycerides under ambient conditions has been a challenge for scientists for many decades. For this reason, so-called extended surfactants were developed that contained hydrophilic/lipophilic linkers to stretch further into the oil and water phase, and enhance the solubility of triglycerides in water. Currently, only limited information about the properties of these surfactants and its behavior in water is available. Therefore, in this work, mixtures of a chosen extended surfactant (C(12-14)-PO(16)-EO(2)-SO(4)Na, X AES) with H(2)O/D(2)O over the whole concentration range were studied by optical microscopy. A schematic phase diagram has been obtained, which shows two isotropic liquid phases at the lowest and highest surfactant concentrations. Furthermore, between the isotropic solutions, four liquid-crystalline phases occur: a hexagonal phase (H(1)), a lamellar phase (L(alpha)) with a change in birefringence, a bicontinuous cubic phase (V(2)), and a reverse hexagonal phase (H(2)). The structure of the micellar solution (L(1)) was determined by cryo-TEM, dynamic light scattering, and (1)H NMR, which gave information about the size, the aggregation number, and the area per molecule of the micelles. Liquid-crystal formation occurs from the micellar solution in two different ways. The first route appeared by increasing the temperature, going from an L(1) to an L(alpha) phase. By increasing the surfactant concentration (at low temperatures), a second route showed a transition from L(1) to H(1). In addition, the effect of sodium chloride on the cloud point of the extended surfactant was examined, indicating that small amounts of NaCl have no influence on the phase behavior. The monolayer behavior of the extended surfactant at the air-water interface was also determined. Despite its water solubility, an isotherm on the water subphase was found, showing slow kinetics of the molecules to go into the bulk. Thus, the determination of the cmc of the extended surfactant using conventional methods was found to be impossible. PMID- 20000428 TI - Interfacial flow processing of collagen. AB - A new method for creating substrates made out of ordered collagen fibers, on which cells in culture can align, is proposed. The substrates can be used for research in cell culture, and this research presents a significant advance in the technology to coat implants in order to improve cell adhesion. In the procedure presented here, a molecular solution of collagen is spread at the interface of a saline solution and air to induce fiber formation, compressed at a high speed to induce orientation and deposited on solid substrates via Langmuir-Blodgett transfer. Several interfacial techniques are employed to investigate the behavior of collagen, which is shown to be dependent on the salt concentration of the subphase as well as the temperature. After Langmuir-Blodgett transfer, primary human fibroblasts and adipose-derived stem cells are cultured on the collagen substrates. Both types of cells respond favorably to the collagen orientation and align with the deposited fibers. The technique presented here provides a simple method to produce well-controlled, oriented collagen substrates that can be used in tissue culture research or scaffolding applications without the use of additives and/or bioincompatible materials. PMID- 20000430 TI - Light-triggered liposomal release: membrane permeabilization by photodynamic action. AB - Photosensitized damage to liposome membranes was studied by using different dye leakage assays based on fluorescence dequenching of a series of dyes upon their release from liposomes. Irradiation of liposomes with red light in the presence of a photosensitizer, trisulfonated aluminum phthalocyanine (AlPcS(3)), resulted in the pronounced leakage of carboxyfluorescein, but rather weak leakage of sulforhodamine B and almost negligible leakage of calcein from the corresponding dye-loaded liposomes. The same series of selectivity of liposome leakage was obtained with chlorin e6 that appeared to be more potent than AlPcS(3) in bringing about the photosensitized liposome leakage. Electrically neutral zinc phthalocyanine tetrasubstituted with a glycerol moiety (ZnPcGlyc(4)) was less effective than negatively charged AlPcS(3) in provoking the light-induced liposome permeabilization. On the contrary, both ZnPcGlyc(4) and AlPcS(3) were much more effective than chlorin e6 in sensitizing gramicidin channel inactivation in planar bilayer lipid membranes, thus showing that relative photodynamic efficacy of sensitizers can differ substantially for damaging different membrane targets. The photosensitized liposome permeabilization was apparently associated with oxidation of lipid double bonds by singlet oxygen as evidenced by the mandatory presence of unsaturated lipids in the membrane composition for the photosensitized liposome leakage to occur and the sensitivity of the latter to sodium azide. The fluorescence correlation spectroscopy measurements revealed marked permeability of photodynamically induced pores in liposome membranes for such photosensitizer as AlPcS(3). PMID- 20000431 TI - Silica-coated gold nanorods with a gold overcoat: controlling optical properties by controlling the dimensions of a gold-silica-gold layered nanoparticle. AB - Silica shells were directly coated onto surfactant-capped gold nanorods by a simple one-step method. The procedure required no intermediate coating of the gold nanorod prior to the formation of the smooth silica shell, the thickness of which could be accurately controlled over the range 60-150 nm. These silica encased gold nanorods were then covered with a gold overcoat to yield nanoparticles with unique optical properties that varied with the thicknesses of both the silica layer and the gold overcoat. Using these bulk solution-phase techniques, homogeneous distributions of gold-silica-gold layered nanoparticles with a pronounced plasmon extinction band in the near-IR (i.e., approximately 900 1700 nm) are readily and reproducibly prepared. More specifically, when using a core gold nanorod whose dimensions yield a plasmon band in the visible region of the spectrum (e.g., approximately 685 nm), the effect of the gold overcoat is to produce a broad plasmon band that is red-shifted by as much as approximately 1000 nm. As such, these multilaminate particles should be of interest as a convenient tool to enhance weak near-IR radiative transitions (e.g., singlet oxygen, O(2)(a(1)Delta(g)), phosphorescence at 1270 nm). PMID- 20000429 TI - Chemical and biological studies of nakiterpiosin and nakiterpiosinone. AB - Nakiterpiosin and nakiterpiosinone are two related C-nor-D-homosteroids isolated from the sponge Terpios hoshinota that show promise as anticancer agents. We have previously described the asymmetric synthesis and revision of the relative configuration of nakiterpiosin. We now provide detailed information on the stereochemical analysis that supports our structure revision and the synthesis of the originally proposed and revised nakiterpiosin. In addition, we herein describe a refined approach for the synthesis of nakiterpiosin, the first synthesis of nakiterpiosinone, and preliminary mechanistic studies of nakiterpiosin's action in mammalian cells. Cells treated with nakiterpiosin exhibit compromised formation of the primary cilium, an organelle that functions as an assembly point for components of the Hedgehog signal transduction pathway. We provide evidence that the biological effects exhibited by nakiterpiosin are mechanistically distinct from those of well-established antimitotic agents such as taxol. Nakiterpiosin may be useful as an anticancer agent in those tumors resistant to existing antimitotic agents and those dependent on Hedgehog pathway responses for growth. PMID- 20000432 TI - SANS from P85/water-d under pressure. AB - Pluronics are triblock copolymers of poly(ethylene oxide)-poly(propylene oxide) poly(ethylene oxide). Pluronic P85 forms a rich phase behavior when dissolved in water. Unimers are observed at low temperature while micelles form at higher temperatures. Spherical, cylindrical, and lamellar micelles are observed sequentially upon heating. The effect of hydrostatic pressure on the phase boundaries is investigated here using small-angle neutron scattering from a dilute P85 solution in deuterated water. The transition temperatures between these micelle phases are found to rise with increasing pressure. A new phase corresponding to demixed lamellae was observed at high temperature. The effect of pressure was seen to decrease the onset formation line of that new phase. PMID- 20000433 TI - Dioxygen activation under ambient conditions: Cu-catalyzed oxidative amidation diketonization of terminal alkynes leading to alpha-ketoamides. AB - A novel Cu-catalyzed oxidative amidation-diketonization reaction of terminal alkynes leading to alpha-ketoamides has been developed. This chemistry offers a valuable mechanistic insight into this novel Cu catalysis via a radical process. O(2) not only participates as the ideal oxidant but also undergoes dioxygen activation under ambient conditions in this transformation. PMID- 20000434 TI - Effect of phospholipase A2 hydrolysis products on calcium oxalate precipitation at lipid interfaces. AB - Urinary stones are commonly composed of an inorganic component, calcium oxalate, or calcium phosphate and an organic matrix of lipids, carbohydrates, and proteinaceous matter. Of interest is the role that the organic matrix elements may play as catalysts for the heterogeneous nucleation of the calcium salts, and a number of studies have examined the role of lipids in calcium oxalate monohydrate (COM) formation. In this study, products of lipid hydrolysis from phospholipase A(2) (PLA(2)) are examined for their effect on COM formation using Langmuir monolayers as model lipid membrane assemblies. The enzyme PLA(2) hydrolyzes DPPC monolayers in the presence of a supersaturated calcium oxalate subphase, inducing the rapid and plentiful nucleation of calcium oxalate at the lipid interface. To investigate the cause of increased crystal formation in the presence of the enzyme, Langmuir monolayers modeling the hydrolysis products were investigated. Calcium oxalate crystal growth at a ternary monolayer of dipalmitoylphosphatidylcholine (DPPC), palmitic acid (PA), and a 22-carbon chain lysophospholipid (22:0 Lyso PC) dramatically increases relative to monolayers of just DPPC. Binary monolayers of DPPC with either PA or the 22:0 Lyso PC and single-component monolayers of PA were also studied. It is demonstrated that the fatty acid generated during lipid hydrolysis causes a significant increase in the extent of heterogeneous nucleation of calcium oxalate from supersaturated solutions. The results imply a possible link between increased phospholipase activity, which is associated with hyperoxaluria, and calcium oxalate precipitation. PMID- 20000435 TI - Comparison of chemical lithography using alkanethiolate self-assembled monolayers on GaAs (001) and Au. AB - We have investigated the efficiency of bifunctional pattern formation in alkanethiolate self-assembled monolayers (SAMs) adsorbed on GaAs (001) and Au, using time-of-flight secondary ion mass spectrometry. Two patterning techniques were employed: electron beam lithography and UV photopatterning. Previous work has always assumed that complete degradation of the SAM was necessary for the formation of well-defined multifunctional patterned surfaces, requiring large electron doses or long UV irradiation times. We demonstrate that well-defined multifunctional patterned surfaces can be produced on GaAs (001) with only partial degradation of the SAM, allowing greatly reduced electron beam doses and UV irradiation times to be used. Using electron beam lithography we observe that sharp well-defined patterns can form after an electron dose as low as 450 microC cm(-2). We also demonstrate that only 50% of the monolayer must be photooxidized in UV photopatterning, reducing the exposure time needed by a factor of 3. In contrast, patterning of alkanethiolate SAMs adsorbed on Au requires much higher electron doses (> or = 1250 microC cm(-2)) and photooxidation times (2 h). The substantial differences observed on these two substrates appear to arise from differences in the SAM structure on GaAs and Au. These results suggest that alkanethiolate SAM resists may be a suitable technology for nanometer scale lithography of GaAs and possibly other semiconductors. PMID- 20000436 TI - Structural motifs and biological studies of new antimony(III) iodide complexes with thiones. AB - Eight new antimony(III) iodide complexes of the heterocyclic thioamides, 2 mercapto-1-methylimidazole (MMI), 2-mercaptobenzimidazole (MBZIM), 5-ethoxy-2 mercaptobenzimidazole (EtMBZIM), 2-mercaptothiazolidine (MTZD), 3-methyl-2 mercaptobenzothiazole (NMeMBZT), 2-mercapto-3,4,5,6-tetrahydropyrimidine (tHPMT), 2-mercaptopyridine (PYT), and 2-mercaptopyrimidine (PMT) of formulas {[SbI(3)(MMI)(2)].MeOH} (1), [SbI(3)(MBZIM)(2)] (2), {[SbI(2)(mu(2) I)(EtMBZIM)(2)](2).H(2)O} (3), [SbI(3)(MTZD)] (4), [(NMeMBZT)SbI(2)(mu(2) I)(2)(mu(2)-S-NMeMBZT)SbI(2) (NMeMBZT)] (5), {[SbI(3)(tHPMT)(3)].MeOH} (6), [SbI(3)(PYT)] (7), and [SbI(3)(PMT)(2)] (8), have been synthesized and characterized by elemental analysis, FT-IR spectroscopy, FT-Raman spectroscopy, and TG-DTA analysis. The crystal structures of 3, 4, 5, 6, and 7 were also determined by X-ray diffraction. The complexes show interesting structural motifs. Complex 6 is a monomer, with octahedral (Oh) geometry around the metal ion formed by three sulfur and three iodide atoms. Complexes 3 and 5 are dimers, with a square pyramidal (SP) geometry in each monomeric unit, while complexes 4 and 7 are polymers with pseudotrigonal bipyramidal (psi-TBP). Two or three sulfur atoms from thioamide ligands and three iodide atoms are bound to Sb atoms forming building blocks for the dimers and polymers. Strong intramolecular interactions between mu(2)-I and/or mu(2)-S and Sb atoms stabilize both structures. In dimer complex 5, two terminal iodide and one terminal sulfur atom are bonded to the Sb ion, while two mu(2)-I and one mu(2)-S bridging atoms bridge the metal ions forming psi-Oh geometry. Computational studies using multivariant linear regression (MLR) and artificial neural networks (ANN) and considering biological results (50% inhibitory concentration, IC(50)) as dependent variables derived a theoretical equation for IC(50) values of the complexes studied. The calculated IC(50) values are compared satisfactorily with the experimental inhibitory activity of the complexes measured. Complexes 3-7 were used to study their influence upon the catalytic peroxidation of linoleic acid by the enzyme Lipoxygenase (LOX). Compounds 1-8 were also tested for in vitro cytotoxicity, and they showed mostly a moderate cytostatic activity against a variety of tumor cell lines but comparable with those found for the antimony(III) chloride and bromide complexes, reported earlier [Ozturk et al. Inorg. Chem. 2007, 46, 2861-2866; Ozturk et al. Inorg. Chem. 2009, 48, 2233-2245]. PMID- 20000437 TI - Vesicle-directed generation of gold nanoflowers by gemini amphiphiles and the spacer-controlled morphology and optical property. AB - In this article, we developed an effective approach to generate gold nanoflowers (AuNFs) by vesicles made from a series of gemini amphiphiles (G2-G10) with different spacer lengths. The gemini amphiphiles were found to form vesicles in aqueous solution. Upon mixing with vesicles in the presence of AgNO(3), HAuCl(4) could be reduced into gold nanoflowers by ascorbic acid. The vesicles directed the growth of the AuNFs, and the spacer length of the gemini amphiphiles showed obvious control of the morphology and optical properties of the formed AuNFs. At a lower HAuCl(4) concentration, the minimum-sized AuNFs were formed when vesicles from the amphiphile with a spacer length of 4 were applied. Upon increasing the spacer length, branched nanoflowers are predominantly produced. A seed-growth mechanism together with the conformational change of the spacer of the gemini amphiphiles was proposed according to the studies on reaction processes. In addition, the formed gold nanoflowers showed obvious surface-enhanced Raman scattering activity for R6G. The present method provided an efficient, controllable way to synthesize branched gold nanostructures. PMID- 20000438 TI - Synthetic fusion peptides of tick-borne encephalitis virus as models for membrane fusion. AB - The fusion peptide of TBEV is a short segment of the envelope protein that mediates viral and host cell membrane fusion at acidic pH. Previous studies on the E protein have shown that mutations at L107 have an effect on fusogenic activity. Structural studies have also suggested that during the fusion process the E protein rearranges to form a trimer. In the present study, a number of short peptides were synthesized, and their structure/activity was examined: (1) monomers consisting of residues 93-113 of the wild-type E protein with Leu at position 107 (WT) and two mutants, namely, L107F and L107T; (2) a monomer consisting of residues 93-113 of the E protein with a C105A mutation (TFPmn); (3) a trimer consisting of three monomers described in (2), linked at the C-terminus via 1 Lys (TFPtr); (4) a monomer consisting of residues 93-113 of the E protein plus six additional Lys at the C-terminus; and (5) a trimer consisting of three monomers described in (3), linked via the side chain of the sixth lysine. The secondary structure content of all peptides was investigated using circular dichroism (CD). Approximately seven of the residues were in beta-strand conformation, in the presence of POPC/POPE/cholesterol. The structures did not depend on pH significantly. The fusogenicity of the peptides was measured by FRET and photon correlation spectroscopy. The data suggest that TFPtr is the most fusogenic at acidic pH and that the mutation from L107 to T reduces activity. Molecular dynamics simulations of WT, L107T, and L107F suggest that this reduction in activity may be related to the fact that the mutations disrupt trimer stability. Finally, tryptophan fluorescence experiments were used to localize the peptides in the membrane. It was found that WT, L107F, TFPmn, and TFPtr could penetrate better into the acyl chain region of the lipids than the other peptides tested. The implications of these results on the fusion mechanism of TBEV E protein will be presented. PMID- 20000439 TI - Nucleation of epitaxial graphene on SiC(0001). AB - A promising route for the synthesis of large-area graphene, suitable for standard device fabrication techniques, is the sublimation of silicon from silicon carbide at elevated temperatures (>1200 degrees C). Previous reports suggest that graphene nucleates along the (110n) plane, known as terrace step edges, on the silicon carbide surface. However, to date, a fundamental understanding of the nucleation of graphene on silicon carbide is lacking. We provide the first direct evidence that nucleation of epitaxial graphene on silicon carbide occurs along the (110n) plane and show that the nucleated graphene quality improves as the synthesis temperature is increased. Additionally, we find that graphene on the (110n) plane can be significantly thicker than its (0001) counterpart and appears not to have a thickness limit. Finally, we find that graphene along the (110n) plane can contain a high density of structural defects, often the result of the underlying substrate, which will undoubtedly degrade the electronic properties of the material. Addressing the presence of non-uniform graphene that may contain structural defects at terrace step edges will be key to the development of a large-scale graphene technology derived from silicon carbide. PMID- 20000440 TI - Synthesis of spiro compounds through tandem oxidative coupling and a framework rearrangement reaction. AB - A highly efficient oxidative coupling of 2-naphthols and a rearrangement tandem reaction to afford unique spiro compounds in the presence of FeCl(3).6H(2)O in up to 88% yield have been developed. PMID- 20000442 TI - Molecular machinery: synthesis of a "nanodragster". AB - The synthesis and imaging by scanning tunneling microscopy of a mixed wheeled nanovehicle composed of a p-carborane small-wheeled short front axle and a C(60) large-wheeled long rear axle that has been termed a nanodragster due to the structural relation to a dragster are reported. This nanodragster is expected to exhibit motion at a lower temperature than pure C(60)-wheeled nanocars and should allow the investigation of the role played by p-carborane wheels in directional motion. PMID- 20000443 TI - Facile synthesis of substituted 5-amino- and 3-amino-1,2,4-thiadiazoles from a common precursor. AB - A novel approach to the synthesis of substituted 5-amino- and 3-amino-1,2,4 thiadiazoles beginning from a common precursor has been achieved. Derivatization by palladium-catalyzed Suzuki-Miyaura coupling enables the rapid preparation of analogs around this pharmaceutically relevant core. FMO calculations rationalize the observed chemoselectivity for coupling at chlorine. PMID- 20000444 TI - Brucine-derived amino alcohol catalyzed asymmetric Henry reaction: an orthogonal enantioselectivity approach. AB - A new approach to both enantioenriched Henry products is developed by use of different molecularities of metal-ligand complexes generated from Cu(I) and Zn(II) metals with readily available chiral amino alcohol 1. PMID- 20000445 TI - Microwave-assisted domino and multicomponent reactions with cyclic acylketenes: expeditious syntheses of oxazinones and oxazindiones. AB - The microwave-assisted Wolff rearrangement of cyclic 2-diazo-1,3-diketones in the presence of aldehydes and primary amines provides a straightforward access to functionalized bi- and pentacyclic oxazinones following an unprecedented three component domino reaction. Alternatively, in the presence of acyl azides, an efficient Curtius/Wolff/hetero-Diels-Alder sequence allows the direct synthesis of oxazindiones. PMID- 20000446 TI - Effect of physicochemical anomalies of soda-lime silicate slides on biomolecule immobilization. AB - Glass microscope slides are considered by many as the substrate of choice for microarray manufacturing due to their amenability to various surface chemistry modifications. The use of silanes to attach various functional groups onto glass slides has provided a versatile tool for the covalent immobilization of many diverse biomolecules of interest. We recently noted a dramatic reduction in biomolecule immobilization efficiency on standard microscope slides prepared using a well-characterized silanization method. A survey of commercial soda-lime slides yielded the surprising result that slides purchased prior to 2008 had superior immobilization efficiencies when compared to those purchased after 2008. Characterization of the slides by X-ray photoelectron spectroscopy (XPS), contact angle measurements, and atomic force microscopy (AFM), revealed a significant correlation (R > 0.9) between magnesium content, surface roughness, and bioimmobilization efficiency. High performance slides had higher magnesium content and higher root-mean-square (rms) roughness (P < 0.005) than slides with lower bioimmobilization efficiencies. Although the exact mechanism of how magnesium content and surface roughness affect silane deposition has not yet been defined, we show that recent changes in the chemical and physical properties of commercial soda-lime slides affect the ability of these slides to be covalently modified. PMID- 20000448 TI - Nanopipet voltammetry of common ions across the liquid-liquid interface. Theory and limitations in kinetic analysis of nanoelectrode voltammograms. AB - Finite element simulations of ion transfer (IT) reactions at the nanopipet supported interface between two immiscible electrolyte solutions (ITIES) were carried out, and the numerical results were generalized in the form of an analytical approximation. The developed theory is the basis of a new approach to kinetic analysis of steady-state voltammograms of rapid IT reactions. Unlike the conventional voltammetric protocol, our approach requires the initial addition of a transferable ion to both liquid phases, i.e., to the filling solution inside a nanopipet and the external solution. The resulting steady-state IT voltammogram comprises two waves corresponding to the ingress of the common ion into the pipet and its egress into the external solution. We demonstrate that both ingress and egress waves are required for characterization of pipet geometry and precise determination of thermodynamic and kinetic parameters for rapid IT reactions. In this way, one can eliminate large uncertainties in kinetic parameters, which are inherent in the previously reported approaches to analysis of nearly reversible steady-state voltammograms of either IT at pipet-supported ITIES or electron transfer at solid electrodes. Numerical simulations also suggest that higher current density at the edge of the nanoscale ITIES increases the significance of electrostatic effects exerted by the charged inner surface of a pipet on IT processes. PMID- 20000449 TI - Kinetic study of rapid transfer of tetraethylammonium at the 1,2 dichloroethane/water interface by nanopipet voltammetry of common ions. AB - Steady-state voltammetry at the pipet-supported liquid/liquid interface has previously been used to measure kinetics of simple and facilitated ion transfer (IT) processes. Recently, we showed that the conventional experimental protocol and data analysis produce large uncertainties in kinetic parameters of rapid IT processes extracted from pipet voltammograms. Here, we used a new mode of nanopipet voltammetry, in which a transferable ion is initially present as a common ion in both liquid phases, and improved methodology for silanization of the outer pipet wall to investigate the kinetics of the rapid transfer of tetraethylammonium (TEA(+)) at the 1,2-dichloroethane/water interface. This reaction was often employed as a model system to check the IT theory. The determined standard rate constant and transfer coefficient of the TEA(+) transfer are compared with previously reported values to demonstrate limitations of conventional nanopipet voltammetry with a transferrable ion present only in one liquid phase. PMID- 20000447 TI - Controlling destiny through chemistry: small-molecule regulators of cell fate. AB - Controlling cell fate is essential for embryonic development, tissue regeneration, and the prevention of human disease. With each cell in the human body sharing a common genome, achieving the appropriate spectrum of stem cells and their differentiated lineages requires the selective activation of developmental signaling pathways, the expression of specific target genes, and the maintenance of these cellular states through epigenetic mechanisms. Small molecules that target these regulatory processes are therefore valuable tools for probing and manipulating the molecular mechanisms by which stem cells self-renew, differentiate, and arise from somatic cell reprogramming. Pharmacological modulators of cell fate could also help remediate human diseases caused by dysregulated cell proliferation or differentiation, heralding a new era in molecular therapeutics. PMID- 20000450 TI - Spin interaction in octahedral zinc complexes of mono- and diradical Schiff and mannich bases. AB - The four Schiff bases 2-tert-butyl-4-methoxy-6-[(pyridin-2 ylmethylimino)methyl]phenol, 2,4-di-tert-butyl-6-[(pyridin-2 ylmethylimino)methyl]phenol, 2-tert-butyl-4-methoxy-6-(quinolin-8 yliminomethyl)phenol, and 2,4-di-tert-butyl-6-(quinolin-8-yliminomethyl)phenol) as well as one Mannich base, N,N',N,N'-bis[(2-hydroxy-3,5-di-tert-butylbenzyl)(2 pyridylmethyl)]ethylenediamine, and their zinc bis-phenolate complexes 1-5, respectively, have been prepared. The complexes 4 and 5 have been characterized by X-ray diffraction crystallography, showing a zinc ion within an octahedral environment, with a cis orientation of the phenolate moieties. 1-5 exhibit in their cyclic voltammetry curves two anodic reversible waves attributable to the successive oxidation of the phenolates into phenoxyl radicals. Bulk electrolysis at ca. +0.1 V affords the zinc-coordinated monophenoxyl radical species (1(*))(+) (5(*))(+) characterized by UV-vis absorption bands at 400-440 nm. The more stable radicals are (3(*))(+) and (4(*))(+) (half-life higher than 90 min at 298 K), likely due to the increased charge delocalization within the quinoline moieties. These species exhibit a significant additional near-IR band (epsilon > 1650 M(-1) cm(-1)) attributed to a CT transition. In the two-electron-oxidized species (1(**))(2+)-(5(**))(2+) the radical spins present a weak magnetic coupling. EPR reveals an antiferromagnetic exchange interaction for (1(**))(2+)-(4(**))(2+), whereas an unusual ferromagnetic exchange coupling is operative in (5(**))(2+). The weak magnitude of experimental |J| values (within the 1-5 cm(-1) range) as well as their sign could be well reproduced by DFT calculations at the B3LYP level. The small energy gap between the ground and the first excited spin states allows us to investigate the zero-field splitting (ZFS) of the triplet by EPR spectroscopy. This parameter is found to be axial for all systems, with |D| values of 0.0163 cm(-1) for (1(**))(2+), 0.0182 cm(-1) for (2(**))(2+), 0.0144 cm(-1) for (3(**))(2+), 0.0160 cm(-1) for (4(**))(2+), and 0.0115 cm(-1) for (5(**))(2+). The trend between experimental ZFS is confirmed by DFT calculations, which give further insight regarding its sign (negative for all the compounds). Lower ZFS values are obtained for (2(**))(2+) compared to (1(**))(2+) (and also for (4(**))(2+) compared to (3(**))(2+)), which can be interpreted by an increased delocalization of the spin density over the methoxy para substituent. Significant spin population on the quinoline also contributes to a lowering of the |D| value, as observed when (3(**))(2+) is compared to (1(**))(2+) (and also when (4(**))(2+) is compared to (2(**))(2+)). PMID- 20000451 TI - An unusual Michael addition of 3,3-dimethoxypropanenitrile to 2-aryl acrylates: a convenient route to 4-unsubstituted 5,6-dihydropyrido[2,3-d]pyrimidines. AB - An unusual Michael addition between 2-aryl-substituted acrylates and 3,3 dimethoxypropanenitrile which leads, depending on the reaction temperature (60 or -78 degrees C, respectively), to a 4-methoxymethylene-substituted 4-cyanobutyric ester or to a 4-dimethoxymethyl 4-cyanobutyric ester is described. These compounds can be subsequently converted to 4-unsubstituted pyrido[2,3 d]pyrimidines upon treatment with a guanidine system under microwave irradiation. PMID- 20000452 TI - Stereostructure reassignment and absolute configuration of isoepitaondiol, a meroditerpenoid from Stypopodium flabelliforme. AB - Careful examination of the published NMR data for isoepitaondiol, a meroditerpenoid from Stypopodium flabelliforme, suggests that its published structure 1 must be revised. On the basis of extensive 1D and 2D NMR studies, we now propose that structure 2, with a trans-anti-trans-anti-cis arrangement fits isoepitaondiol diacetate. The relative configuration of 2 was confirmed by single crystal X-ray diffraction, while the absolute configuration was evidenced by vibrational circular dichroism in combination with DFT B3LYP/DGDZVP calculations. PMID- 20000453 TI - Enzymatic production of (-)-indolactam V by LtxB, a cytochrome P450 monooxygenase. AB - The P450 cytochrome monooxygenase gene, ltxB, was cloned and overexpressed in Escherichia coli as a 6xHis-tagged protein. The resulting recombinant LtxB was purified by Ni-NTA affinity chromatography and characterized biochemically. Purified LtxB demonstrated typical cytochrome P450 spectroscopic properties including substrate-induced transition from a low-spin (lambdamax=414 nm) to high spin state (lambdamax=386 nm) upon incubation with N-methyl-L-valyl-L tryptophanol. The catalytic activity of LtxB was verified by demonstrating the oxidation/cyclization of N-methyl-L-valyl-L-tryptophanol to (-)-indolactam V. LtxB shows a relaxed specificity for analogue substrates in which the valyl group is substituted for other aliphatic groups. The relaxed substrate specificity of LtxB, along with the relaxed specificity of the prenyltransferase, LtxC, allowed for the enzymatic production of a series of (-)-indolactam V and lyngbyatoxin analogues. PMID- 20000454 TI - Schweinfurthins I and J from Macaranga schweinfurthii. AB - Demand for the experimental antineoplastic agent schweinfurthin A, for developmental testing, prompted a re-collection of leaf material of Macaranga schweinfurthii from the original collection site in Cameroon. During chromatographic purification of the organic solvent extract, analytical UPLC-PDA TOFMS of stilbene-enriched fractions revealed the presence of six known schweinfurthins and two previously unknown stilbenes. The structures of these new compounds, schweinfurthins I and J (1 and 2), were elucidated by 1D- and 2D-NMR techniques. PMID- 20000455 TI - Direct binding of thymopentin to surface class II major histocompatibility complex in living cells. AB - The molecular analysis of thymopentin (TP5)/class II major histocompatibility complex (MHC II) complexes has been basically understood; however, the mechanism by which TP5-MHC II complexes are formed is largely unexplored. Compared with Epstein-Barr virus (EBV)-transformed B cells expressing human leucocyte antigen DR (HLA-DR), no fluorescent signal was observed on the DR-deficient cell line. This indicates that FITC labeled TP5 (FITC-TP5) is genuinely bound to HLA-DR. The binding specificity was confirmed by incubating FITC-TP5 with unlabeled TP5 and HA peptide as well as mAb for DR molecules. In addition, the binding appeared to be rapid in living antigen-presenting cell (APC), which implies that TP5 is demonstrated on APC surface and does not require processing before associating with DR. Additional support for this surface binding arises from the observation that pretreatment of cells with a variety of metabolic inhibitors failed to decrease the level of TP5/DR complexes. However, temperature has an effect on the rate of binding between TP5 and DR molecules, which is well consistent with the qualitative predication of transition state theory. The formation of antigenic complexes is accelerated at acidic pH, which shows that the formation of TP5/DR complexes is a pH-dependent process. PMID- 20000456 TI - Negative dissociation energy phenomenon of metastable H-bonds as revealed in triplex DNA hole migration. AB - Ab initio calculations reveal an unknown energetic phenomenon for H-bonds in the hole-trapping triplex C(p)*GC motif observed experimentally in hole migration which can explain the lower but really available oxidization possibility in C(p)*GC site. Hole trapping can considerably destabilize the C(p)*GC unit and lead to an unexpected barrier-hindered channel with a negative dissociation energy. This channel is governed by a balance between electrostatic repulsion and H-bonding attraction in the two associated moieties and different attenuations of two opposite interactions with respect to the H-bond distance. This C(p)*GC unit can be viewed as a high-energy node in a DNA wire which modulates migration of a hole into or through it via its unusual energetics. It provides useful information for understanding of an unknown type of the complicated intermolecular interactions, a novel type of "high-energy" bond, and can be applied further to interpret the hidden transport properties and the energy conversion/transfer mechanisms in the related fields. PMID- 20000457 TI - Exploiting binding-induced changes in probe flexibility for the optimization of electrochemical biosensors. AB - Electrochemical sensors employing redox-tagged, electrode-bound oligonucleotides have emerged as a promising new platform for the reagentless detection of molecular analytes. Signal generation in these sensors is linked to specific, binding-induced changes in the efficiency with which an attached redox tag approaches and exchanges electrons with the interrogating electrode. We present here a straightforward means of optimizing the signal gain of these sensors that exploits this mechanism. Specifically, using square-wave voltammetry, which is exquisitely sensitive to electrode reaction rates, we can tune the frequency of the voltammetric measurements to preferentially enhance the signal associated with either the unbound or target-bound conformations of the probe. This allows us to control not only the magnitude of the signal gain associated with target binding but also the sign of the signal change, generating "signal-on" or "signal off" sensors. This optimization parameter appears to be quite general: we show here that tuning the square-wave frequency can significantly enhance the gain of the sensors directed against specific oligonucleotide sequences, small molecules, proteins, and protein-small molecule interactions. PMID- 20000458 TI - Controlled synthesis of camptothecin-polylactide conjugates and nanoconjugates. AB - We report here a unique method of formulating camptothecin-polylactide (CPT-PLA) conjugate nanoparticles, termed nanoconjugates (NCs), through CPT/(BDI)ZnN(TMS)(2) [(BDI) = 2-((2,6-diisopropylphenyl)amido)-4-((2,6-bisalkyl) imino)-2-pentene] mediated polymerization of lactide (LA) followed by nanoprecipitation. When CPT was used as the initiator to polymerize LA in the presence of (BDI)ZnN(TMS)(2), the polymerization was completed within hours with nearly 100% CPT loading efficiency and 100% LA conversion. CPT loading as high as 19.5% can be achieved for the CPT-polylactide (CPT-PLA) conjugate prepared at a LA/CPT ratio of 10. The steric bulk of the chelating ligands and the type of metals used had a dramatic effect on the initiation of the LA polymerization and the tendency of the ring-opening of the CPT lactone. The CPT/(BDI)ZnN(TMS)(2) mediated LA polymerization yielded CPT-PLA conjugates with well-controlled molecular weights and narrow molecular weight distributions (1.02-1.18). The nanoprecipitation of CPT-PLA led to the formation of NCs around 100 nm in size with narrow particle size distributions. Sustained release of CPT from CPT-PLA NCs was achieved without burst release. CPT-PLA NCs were toxic to PC-3 cells with tunable IC(50) possible by adjusting the drug loading of the CPT-PLA NCs. PMID- 20000459 TI - Modification of thiol functionalized aptamers by conjugation of synthetic polymers. AB - Aptamers are known for their short in vivo circulating half-life and rapid renal clearance. Their conjugation to poly(ethylene glycol) (PEG) is a way to improve their residence in the body. Two aptamers (AptD and AptF), having a disulfide protected thiol modification on the 3' end, have been conjugated to maleimide activated PEGs of various molecular weights and structures (linear PEG20; branched PEG20 and 40; PolyPEG17, 40, and 60 kDa). The high yield coupling (70 80% in most of the cases) could be achieved using immobilized tris[2 carboxyethyl]phosphine hydrochloride (TCEP) as reducing agent at pH 4. The affinity of PEGylated AptD for its target was reduced by conjugation to linear PEG20 and branched PEG40, but not to branched PEG20 and PolyPEGs. This work demonstrates an alternative approach to PEGylation of aptamers, and that the effect of PEG on the affinity for the target varies according to the structure and conformation of the synthetic polymer. PMID- 20000460 TI - Two-step biocatalytic route to biobased functional polyesters from omega-carboxy fatty acids and diols. AB - Biobased omega-carboxy fatty acid monomers 1,18-cis-9-octadecenedioic, 1,22-cis-9 docosenedioic, and 1,18-cis-9,10-epoxy-octadecanedioic acids were synthesized in high conversion yields from oleic, erucic and epoxy stearic acids by whole-cell biotransformations catalyzed by C. tropicalis ATCC20962. Maximum volumetric yields in shake-flasks were 17.3, 14.2, and 19.1 g/L after 48 h conversion for oleic acid and 72 h conversions for erucic and epoxy stearic acids, respectively. Studies in fermentor with better control of pH and glucose feeding revealed that conversion of oleic acid to 1,18-cis-9-octadecenedioic acid by C. tropicalis ATCC20962 occurred with productivities up to 0.5 g/L/h. The conversion of omega carboxy fatty acid monomers to polyesters was then studied using immobilized Candida antarctica Lipase B (N435) as catalyst. Polycondensations with diols were performed in bulk as well as in diphenyl ether. The retension of functionality from fatty acid, to omega-carboxy fatty acid monomer and to corresponding polyesters resulted in polymers with with unsaturated and epoxidized repeat units and M(w) values ranging from 25000 to 57000 g/mol. These functional groups along chains disrupted crystallization giving materials that are low melting (23-40 degrees C). In contrast, saturated polyesters prepared from 1,18-octadecanedioic acid and 1,8-octanediol have correspondingly higher melting transitions (88 degrees C). TGA results indicated that all synthesized polyesters showed high thermal stabilities. Thus, the preparation of functional monomers from C. tropicalis omega-oxidation of fatty acids provides a wide range of new monomer building blocks to construct functional polymers. PMID- 20000461 TI - Protein-resistant polymer coatings based on surface-adsorbed poly(aminoethyl methacrylate)/poly(ethylene glycol) copolymers. AB - We report on the protein-resistant properties of glass substrates coated with novel copolymers of 2-aminoethyl methacrylate hydrochloride and poly(ethylene glycol) methyl ether methacrylate (AEM-PEG). In comparison to currently available protein-blocking polymer systems, such as poly-l-lysine-poly(ethylene glycol), silane-based poly(ethylene glycol), and poly(ethylene glycol) brushes prepared by surface-initiated polymerization, the proposed AEM-PEG offers the combined advantages of low cost, simplicity of use, and applicability in aqueous solutions. We demonstrate the capability of AEM-PEG to block the surface binding of globular proteins (tubulin), their assemblies (microtubules), and functional motor proteins (kinesin-1). Moreover, we demonstrate the applicability of AEM-PEG for surface patterning of proteins in microfluidic devices. PMID- 20000462 TI - Effect of glass dissolution on the solution deposition of ZnO films and its exploitation for deposition of Zn silicates. AB - ZnO is probably the most studied material deposited as films by aqueous solution methods. Both neutral and alkaline solutions are commonly used, and deposition is often carried out in glass vessels. We show that for depositions carried out under alkaline conditions, slow dissolution of the glass by the solution often results in formation of zinc silicates together with the ZnO. While this silicate formation is most clearly seen after long deposition times (many hours), it can be detected already within 1 h, while often ZnO depositions proceed for substantially longer. We also describe conditions where the zinc silicate deposits without formation of ZnO providing a method of depositing such films. Finally, we note that the glass of a reaction vessel also can affect deposition of CdSe, pointing to a more general role of this normally neglected parameter. PMID- 20000463 TI - Extraction of phenolic acids by alkaline hydrolysis from the solid residue obtained after prehydrolysis of trimming vine shoots. AB - Contents of hydroxycinnamic and hydroxybenzoic acids were determined in trimming vine shoots after sequential treatments of prehydrolysis and alkaline hydrolysis. These treatments allow the complete use of the main fractions involved: cellulose, hemicelluloses and lignin. The alkaline hydrolysis was studied using a factorial design where reaction time (in the range 30-120 min), temperature (50 130 degrees C), and NaOH concentration (4-12 wt % of solution) were the independent variables. The interrelationship between dependent and operational variables was well fitted (R(2) > 0.90) to models including linear, interaction and quadratic terms. Ferulic acid was the most abundant hydroxycinnamate with concentrations ranging from 25.7 to 141.0 mg/L followed by p-coumaric acid (15.5 31.5 mg/L). Gallic acid was the hydroxybenzoic acid released in higher concentration (in the range 2.5-164.6 mg/L). Because of their properties and low toxicity, these compounds are widely used in the food, pharmaceutical and cosmetic industries. Additionally, ferulic acid is used as feedstock for the biotechnological production of flavorings and aroma compounds, including vanillin and vinylguaiacol, or as a constituent in the preparation of foods and skin protection agents, or as a cross-linking agent for the elaboration of food gels. Consequently, ferulic acid solutions can be obtained from renewable plant cell wall materials as a prospective pathway. PMID- 20000464 TI - Strongly nonlinear optical glass fibers from noncentrosymmetric phase-change chalcogenide materials. AB - We report that the one-dimensional polar selenophosphate compounds APSe(6) (A = K, Rb), which show crystal-glass phase-change behavior, exhibit strong second harmonic generation (SHG) response in both crystal and glassy forms. The crystalline materials are type-I phase-matchable with SHG coefficients chi((2)) of 151.3 and 149.4 pm V(-1) for K(+) and Rb(+) salts, respectively, which is the highest among phase-matchable nonlinear optical (NLO) materials with band gaps over 1.0 eV. The glass of APSe(6) exhibits comparable SHG intensities to the top infrared NLO material AgGaSe(2) without any poling treatments. APSe(6) exhibit excellent mid-IR transparency. We demonstrate that starting from noncentrosymmetric phase-change materials such as APSe(6) (A = K, Rb), we can obtain optical glass fibers with strong, intrinsic, and temporally stable second order nonlinear optical (NLO) response. The as-prepared glass fibers exhibit SHG and difference frequency generation (DFG) responses over a wide range of wavelengths. Raman spectroscopy and pair distribution function (PDF) analyses provide further understanding of the local structure in amorphous state of KPSe(6) bulk glass and glass fiber. We propose that this approach can be widely applied to prepare permanent NLO glass from materials that undergo a phase-change process. PMID- 20000465 TI - Solution-processed inorganic solar cell based on in situ synthesis and film deposition of CuInS2 nanocrystals. AB - A novel, reproducible, and simple solution-based process for the fabrication of CuInS(2) absorber layers and CdS buffer layers for photovoltaics is presented. In this process, a precursor solution is deposited on a substrate, after which sintered NCs are formed in situ at temperatures as low as 250 degrees C. Solar cell efficiencies of up to 4% have been demonstrated using this novel fabrication method. PMID- 20000466 TI - Detection of a transient intermediate in a rapid protein folding process by solid state nuclear magnetic resonance. AB - We describe the use of solid-state NMR spectroscopy to characterize a partially folded state of the 35-residue helical protein HP35 created by rapid freeze quenching from a thermally unfolded state on the 10-20 micros time scale. Two dimensional solid-state (13)C NMR spectra of (13)C-labeled HP35 in frozen glycerol/water solution exhibit two sets of signals, one corresponding to strongly unfolded protein molecules and the other to an ensemble of molecules having native helical secondary structure but incomplete tertiary structure. The NMR data indicate that secondary structure forms within the freeze-quenching time scale but that full folding involves a slower phase of structural annealing. The approximately 5 micros folding time observed in earlier studies of HP35 by time resolved optical techniques may not represent the time scale for full folding. PMID- 20000467 TI - Regulation of succinate dehydrogenase activity by SIRT3 in mammalian mitochondria. AB - A member of the sirtuin family of NAD(+)-dependent deacetylases, SIRT3, is identified as one of the major mitochondrial deacetylases located in mammalian mitochondria responsible for deacetylation of several metabolic enzymes and components of oxidative phosphorylation. Regulation of protein deacetylation by SIRT3 is important for mitochondrial metabolism, cell survival, and longevity. In this study, we identified one of the Complex II subunits, succinate dehydrogenase flavoprotein (SdhA) subunit, as a novel SIRT3 substrate in SIRT3 knockout mice. Several acetylated Lys residues were mapped by tandem mass spectrometry, and we determined the role of acetylation in Complex II activity in SIRT3 knockout mice. In agreement with SIRT3-dependent activation of Complex I, we observed that deacetylation of the SdhA subunit increased the Complex II activity in wild-type mice. In addition, we treated K562 cell lines with nicotinamide and kaempferol to inhibit deacetylase activity of SIRT3 and stimulate SIRT3 expression, respectively. Stimulation of SIRT3 expression decreased the level of acetylation of the SdhA subunit and increased Complex II activity in kaempherol-treated cells compared to control and nicotinamide-treated cells. Evaluation of acetylated residues in the SdhA crystal structure from porcine and chicken suggests that acetylation of the hydrophilic surface of SdhA may control the entry of the substrate into the active site of the protein and regulate the enzyme activity. Our findings constitute the first evidence of the regulation of Complex II activity by the reversible acetylation of the SdhA subunit as a novel substrate of the NAD(+)-dependent deacetylase, SIRT3. PMID- 20000468 TI - Studies on the mechanism of p-hydroxyphenylacetate 3-hydroxylase from Pseudomonas aeruginosa: a system composed of a small flavin reductase and a large flavin dependent oxygenase. AB - There are two known types of microbial two-component flavin-dependent monooxygenases that catalyze oxygenation of p-hydroxyphenylacetate (HPA), and they are distinguished by having structurally distinct reductases and oxygenases. This paper presents a detailed analysis of the properties of the enzyme from Pseudomonas aeruginosa, an example of one group, and compares its properties to those published for the Acinetobacter baumannii enzyme, an example of the alternative group. The reductase and oxygenase from P. aeruginosa were expressed in Escherichia coli. The reductase was purified as a stable C-terminally His tagged yellow protein containing weakly bound FAD, and the oxygenase was purified as a stable colorless N-terminally His-tagged protein. The reductase catalyzes the reduction of FAD by NADH and releases the FADH(-) product into solution, but unlike the reductase from A. baumannii, this catalysis is not influenced by HPA. The oxygenase binds the released FADH(-) and catalyzes the oxygenation of HPA to form 3,4-dihydroxyphenylacetate, after which the FAD dissociates to be re-reduced by the reductase, a common overall pattern for two-component flavin-dependent oxygenases. With this system, it appears that interactions between the reductase and the oxygenase can facillitate the transfer of FADH(-) to the oxygenase, although they are not required. We show that the P. aeruginosa oxygenase system in complex with FADH(-) reacts with O(2) to form a quasi-stable, unusually high extinction flavin hydroperoxide species that binds HPA and reacts to form the product. The resultant flavin hydroxide decomposes to FAD and water while still bound to the oxygenase and then releases product and FAD from the protein. Unlike the enzyme from A. baumannii, during normal catalysis involving both the reductase and oxygenase, the rate-determining step in catalysis is the dissociation of FAD from the oxygenase in a process that is independent of the concentration of HPA. Structures for the reductases and oxygenases from A. baumannii and from Thermus thermophilus (similar to the P. aeruginosa system) form a basis for interpreting the molecular origins of the differences between the two groups of flavin-dependent two-component oxygenases. PMID- 20000469 TI - Hit to lead account of the discovery of bisbenzamide and related ureidobenzamide inhibitors of Rho kinase. AB - A highly selective series of bisbenzamide inhibitors of Rho-associated coiled coil forming protein kinase (ROCK) and a related ureidobenzamide series, both identified by high throughput screening (HTS), are described. Details of the hit validation and lead generation process, including structure-activity relationship (SAR) studies, a selectivity assessment, target-independent profiling (TIP) results, and an analysis of functional activity using a rat aortic ring assay are discussed. PMID- 20000470 TI - Identification, synthesis, and pharmacological evaluation of tetrahydroindazole based ligands as novel antituberculosis agents. AB - The resurgence of tuberculosis (TB), the incidence of drug-resistant strains of Mycobacterium tuberculosis (MTB), and the coinfection between TB and HIV have led to serious infections, high mortality, and a global health threat, resulting in the urgent search for new classes of antimycobacterial agents. Herein, we report the identification of a novel class of tetrahydroindazole based compounds as potent and unique inhibitors of MTB. Compounds 6a, 6m, and 6q exhibited activity in the low micromolar range against replicating Mycobacterium tuberculosis (R-TB) phenotype, with minimum inhibitory concentrations (MICs) of 1.7, 1.9, and 1.9 muM, respectively, while showing no toxicity to Vero Ccells. Moreover, studies aimed to assess the in vitro metabolic stability of 6a and 6m in mouse liver microsomes and in vivo pharmacokinetic profiles in plasma levels gave satisfactory results. This research suggests that tetrahydroindazole based anti TB compounds can serve as a promising lead scaffold in developing new drugs to combat tuberculosis infections. PMID- 20000471 TI - One-pot facile synthesis of pyridyl annelated perylene bisimides. AB - Two regiospecifically pyridyl annelated perylene bisimides have been prepared in one pot by the combination of the Suzuki cross-coupling reaction and subsequent light-promoted cyclization in high yields. PMID- 20000472 TI - Chemical structure determination of DNA bases modified by active metabolites of lucidin-3-O-primeveroside. AB - Lucidin-3-O- primeveroside (LuP) is one of the components of madder root (Rubia tinctorum L.; MR) which is reported to be carcinogenic in the kidney and liver of rats. Since metabolism of LuP generates genotoxic compounds such as lucidin (Luc) and rubiadin (Rub), it is likely that LuP plays a key role in MR carcinogenesis. In the present study, the chemical structures of Luc-specific 2'-deoxyguanosine (dG) and 2'-deoxyadenosine (dA) adducts following the reactions of dG and dA with a Luc carbocation or quinone methide intermediate derived from Acetoxy-Luc were determined by liquid chromatography with photodiode array and electron spray ionizaion-mass spectrometry (LC-PDA-ESI/MS). The identification of the two measurable adducts as Luc-N(2)-dG and Luc-N(6)-dA was confirmed by NMR analysis. Subsequently, using a newly developed quantitative analytical method using LC ESI/MS, the formation of Luc-N(2)-dG and Luc-N(6)-dA from the reaction of calf thymus DNA with Luc in the presence of S9 mixture was observed. The fact that this reaction with Rub also gave rise to the same dG and dA adducts strongly suggests that Rub genotoxicity involves a metabolic conversion to Luc. The precise determination of the modified DNA bases generated by LuP and the method for their analysis may contribute to further comprehension of the mode of action underlying carcinogenesis by MR and related anthraquinones. PMID- 20000473 TI - Comparative genotoxicity and cytotoxicity of four hexavalent chromium compounds in human bronchial cells. AB - Hexavalent chromium (Cr(VI)) compounds are well-established human lung carcinogens. Solubility plays an important role in their carcinogenicity with the particulate Cr(VI) compounds being the most carcinogenic. Epidemiology and animal studies suggest that zinc chromate is the most potent particulate Cr(VI) compound; however, there are few comparative data to support these observations. The purpose of this study was to compare the genotoxicity of zinc chromate with two other particulate Cr(VI) compounds, barium chromate and lead chromate, and one soluble Cr(VI) compound, sodium chromate. The clastogenic effects of barium chromate and zinc chromate were similar, but lead chromate induced significantly less damage. The levels of DNA damage measured by gamma-H2A.X foci formation were similar for the three particulate chromium compounds. Corrected for chromium uptake differences, we found that zinc chromate and barium chromate were the most cytotoxic, and lead chromate and sodium chromate were less cytotoxic. Zinc chromate was more clastogenic than all other chromium compounds, and lead chromate was the least clastogenic. There was no significant difference between any of the compounds for the induction of DNA double strand breaks. All together, these data suggest that the difference in the carcinogenic potency of zinc chromate over the other chromium compounds is not due solely to a difference in chromium ion uptake and that the zinc cation may in fact have an important role in its carcinogenicity. PMID- 20000474 TI - Generation of 2'-deoxyadenosine N6-aminyl radicals from the photolysis of phenylhydrazone derivatives. AB - Nitrogen-centered radicals are major species generated by the addition of hydroxyl radicals and the one-electron oxidation of adenine derivatives. Aminyl radicals are also generated in the decomposition of adenine chloramines upon reaction of hypochlorite. Here, we report the photochemistry of modified 2' deoxyadenosine (dAdo) containing photoactive hydrazone substituents as a model to investigate the chemistry of dAdo N(6)-aminyl radicals. Derivatives of dAdo containing a phenylhydrazone moiety at N6 displayed UV absorption between 300 and 400 nm. Upon UV photolysis in the presence of a H-donor, that is, glutathione, two major products were formed, dAdo and benzaldehyde, indicating efficient homolytic cleavage to dAdo N(6)-aminyl radicals and benzylidene iminyl radicals. dAdo N(6)-phenylhydrazone was photolyzed in the presence of a molar excess of nonmodified dAdo to mimic the reactions taking place in DNA, and the major photoproducts were identified by high-performance liquid chromatography, mass spectrometry, and nuclear magnetic resonance. The formation of 2 (benzylideneamino)-2'-deoxyadenosine as well as a more extensive oxidation product may be explained by the recombination of initial dAdo N(6)-aminyl and benzylidene iminyl radicals. The formation of 2'-deoxyinosine may be explained by hydrolytic deamination of dAdo N(6)-aminyl radicals. Interestingly, a dimeric product containing two dAdo moieties was identified in the photolysis mixture. The present studies demonstrate the ability of dAdo N(6)-aminyl radicals to undergo H-abstraction to give dAdo, deamination to give 2'-deoxyinosine, and addition to the adenine moiety to give dimers. PMID- 20000475 TI - Depleted uranium induces neoplastic transformation in human lung epithelial cells. AB - Depleted uranium (DU) is commonly used in military armor and munitions, and thus, exposure of soldiers and noncombatants is frequent and widespread. Previous studies have shown that DU has both chemical and radiological toxicity and that the primary route of exposure of DU to humans is through inhalation and ingestion. However, there is limited research information on the potential carcinogenicity of DU in human bronchial cells. Accordingly, we determined the neoplastic transforming ability of particulate DU to human bronchial epithelial cells (BEP2D). We observed the loss of contact inhibition and anchorage independent growth in cells exposed to DU after 24 h. We also characterized these DU-induced transformed cell lines and found that 40% of the cell lines exhibit alterations in plating efficiency and no significant changes in the cytotoxic response to DU. Cytogenetic analyses showed that 53% of the DU-transformed cell lines possess a hypodiploid phenotype. These data indicate that human bronchial cells are transformed by DU and exhibit significant chromosome instability consistent with a neoplastic phenotype. PMID- 20000476 TI - Suppression of p53 and p21CIP1/WAF1 reduces arsenite-induced aneuploidy. AB - Aneuploidy and extensive chromosomal rearrangements are common in human tumors. The role of DNA damage response proteins p53 and p21(CIP1/WAF1) in aneugenesis and clastogenesis was investigated in telomerase immortalized diploid human fibroblasts using siRNA suppression of p53 and p21(CIP1/WAF1). Cells were exposed to the environmental carcinogen sodium arsenite (15 and 20 microM), and the induction of micronuclei (MN) was evaluated in binucleated cells using the cytokinesis-block assay. To determine whether MN resulted from missegregation of chromosomes or from chromosomal fragments, we used a fluorescent in situ hybridization with a centromeric DNA probe. Micronuclei were predominantly of clastogenic origin in control cells regardless of p53 or p21(CIP1/WAF1) expression. MN with centromere signals in cells transfected with NSC siRNA or Mock increased 30% after arsenite exposure, indicating that arsenite induced aneuploidy in the tGM24 cells. Although suppression of p53 increased the fraction of arsenite-treated cells with MN, it caused a decrease in the fraction with centromeric DNA. Suppression of p21(CIP1/WAF1) like p53 suppression decreased the fraction of MN with centromeric DNA. Our results suggest that cells lacking normal p53 function cannot become aneuploid because they die by mitotic arrest associated apoptosis, whereas cells with normal p53 function that are able to exit from mitotic arrest can become aneuploid. Furthermore, our current results support this role for p21(CIP1/WAF1) since suppression of p21(CIP1/WAF1) caused a decrease in aneuploidy induced by arsenite, suggesting that p21(CIP1/WAF1) plays a role in mitotic exit. PMID- 20000477 TI - The prosegment catalyzes pepsin folding to a kinetically trapped native state. AB - Investigations of irreversible protein unfolding often assume that alterations to the unfolded state, rather than the nature of the native state itself, are the cause of the irreversibility. However, the present study describes a less common explanation for the irreversible denaturation of pepsin, a zymogen-derived aspartic peptidase. The presence of a large folding barrier combined with the thermodynamically metastable nature of the native state, the formation of which depends on a separate prosegment (PS) domain, is the source of the irreversibility. Pepsin is unable to refold to the native state upon return from denaturing conditions due to a large folding barrier (24.6 kcal/mol) and instead forms a thermodynamically stable, yet inactive, refolded state. The native state is kinetically stabilized by an unfolding activation energy of 24.5 kcal/mol, comparable to the folding barrier, indicating that native pepsin exists as a thermodynamically metastable state. However, in the presence of the PS, the native state becomes thermodynamically stable, and the PS catalyzes pepsin folding by stabilizing the folding transition state by 14.7 kcal/mol. Once folded, the PS is removed, and the native conformation exists as a kinetically trapped state. Thus, while PS-guided folding is thermodynamically driven, without the PS the pepsin energy landscape is dominated by kinetic barriers rather than by free energy differences between native and denatured states. As pepsin is the archetype of a broad class of aspartic peptidases of similar structure and function, and many require their PS for correct folding, these results suggest that the occurrence of native states optimized for kinetic rather than thermodynamic stability may be a common feature of protein design. PMID- 20000478 TI - Sulfur-assisted five-cascade sequential reactions for the convenient and efficient synthesis of allyl thiophen-2-yl acetates, propionates, and ketones. AB - A sulfur-assisted five-cascade sequential reaction, wherein the in situ-generated allenyl allyl sulfides undergo thio-Claisen rearrangement, intramolecular Michael addition, and 1, 5-proton migration/aromatization to obtain allyl thiophen-2-yl acetates, propionates, and ketones as the final products, was reported. As a result of the ready availability of starting materials and the extremely simple and convenient operation, this type of reaction presented here has potential utility in organic synthesis. Application of this efficient method for the synthesis of potentially pharmaceutical compounds also might be useful for the pharmacists. PMID- 20000480 TI - Anion photoelectron spectroscopy of C3N- and C5N-. AB - Anion photoelectron spectroscopy of C(3)N(-) and C(5)N(-) is performed using slow electron velocity-map imaging (SEVI) and field-free time-of-flight (TOF), respectively. The SEVI spectrum exhibits well-resolved vibrational transitions from the linear C(3)N(-) ground state to the corresponding C(3)N ground state. The TOF spectrum comprises transitions arising from the linear C(5)N(-) ground state to the corresponding neutral ground and excited states. This study yields the adiabatic electron affinities of C(3)N and C(5)N to be 4.305 +/- 0.001 and 4.45 +/- 0.03 eV, respectively, and a term value of 560 +/- 120 cm(-1) for the A(2)Pi state of C(5)N. Vibrational frequencies for the degenerate cis and trans bending modes of C(3)N(-) are also extracted. PMID- 20000479 TI - Role of signature lysines in the deviant walker a motifs of the ArsA ATPase. AB - The ArsA ATPase belongs to the P-loop GTPase subgroup within the GTPase superfamily of proteins. Members of this subgroup have a deviant Walker A motif which contains a signature lysine that is predicted to make intermonomer contact with the bound nucleotides and to play a role in ATP hydrolysis. ArsA has two signature lysines located at positions 16 and 335. The role of Lys16 in the A1 half and Lys335 in the A2 half was investigated by altering the lysines individually to alanine, arginine, leucine, methionine, glutamate, and glutamine by site-directed mutagenesis. While Lys16 mutants show similar resistance phenotypes as the wild type, the Lys335 mutants are sensitive to higher concentrations of arsenite. K16Q ArsA shows 70% of wild-type ATPase activity while K335Q ArsA is inactive. ArsA is activated by binding of Sb(III), and both wild-type and mutant ArsAs bind Sb(III) with a 1:1 stoichiometry. Although each ArsA binds nucleotide, the binding affinity decreases in the order wild type > K16Q > K335Q. The results of limited trypsin digestion analysis indicate that both wild type and K16Q adopt a similar conformation during activated catalysis, whereas K335Q adopts a conformation that is resistant to trypsin cleavage. These biochemical data along with structural modeling suggest that, although Lys16 is not critical for ATPase activity, Lys335 is involved in intersubunit interaction and activation of ATPase activity in both halves of the protein. Taken together, the results indicate that Lys16 and Lys335, located in the A1 and A2 halves of the protein, have different roles in ArsA catalysis, consistent with our proposal that the nucleotide binding domains in these two halves are functionally nonequivalent. PMID- 20000481 TI - Theoretical study of the structural and electronic properties of the Fe(6) (C(6)H(6))(m), m = 3, 4, complexes. AB - The adsorption of benzene on the magnetic Fe(6) cluster was studied by means of first principles all-electron calculations done with gradient corrected density functional theory. In the M = 2S + 1 = 13 (S is the total spin) ground state (GS) of Fe(6)-(C(6)H(6))(3) each benzene is bonded with one Fe atom, forming eta(6) coordinations with C-Fe contacts of 2.12-2.17 A; though the Fe(6) cluster structure is preserved, it presents more distortion than in bare Fe(6). The M = 13 GS of Fe(6)-(C(6)H(6))(4) shows a more distorted geometry with three eta(6) and one eta(2) coordinations, as the bonding with the fourth benzene was reduced to two C-Fe bonds. Thus, Fe(6)-(C(6)H(6))(4) may be viewed as a Fe(6) core covered by a layer of benzene molecules. The d-pi bonding interactions are clearly reflected by the estimated adiabatic ionization energies (4.60 and 4.42 eV for m = 3 and 4, respectively), because they are significantly smaller than that of bare Fe(6), 6.15 eV. The adiabatic electron affinities also are diminished clearly, 1.02 and 1.13 eV, for m = 3 and 4, respectively, as compared to that of Fe(6), 1.61 eV. The magnetic moments of the Fe(6)-(C(6)H(6))(3,4) complexes are strongly quenched, by 8.0 magneton bohrs (mu(B)), with respect to the value, 20.0 mu(B), of the isolated Fe(6) cluster. Lastly, the vibrational spectra show IR bands placed near those of free benzene and several forbidden IR modes of benzene turn IR active in the reduced symmetry of the complexes. PMID- 20000482 TI - Quantum-chemical studies on hexaazaisowurtzitanes. AB - Highly nitrated cage molecules constitute a new class of energetic materials that have received a substantial amount of interest. Among them 2,4,6,8,10,12 hexanitro-2,4,6,8,10,12-hexaazaisowurtzitane (CL-20) is a powerful explosive with poor impact and friction characteristics. In the present study we aim to design novel energetic materials by tailoring the molecular structure of CL-20. Important characteristics such as the heat of formation and density have been predicted using density functional theory and packing calculations, respectively. Sensitivity correlations have been established for model compounds by analyzing the charge on the nitro groups. Molecules IDX1, IDX4, and IDX7 have been found to have comparable performance with better insensitivity characteristics and may be explored as CL-20 substitutes in defense applications. PMID- 20000483 TI - DFT/TD-DFT investigation of electronic structures and spectra properties of Cu based dye sensitizers. AB - Molecular geometries, electronic structures, and optical absorption spectra were investigated using density functional theory (DFT) at the B3LYP/6-31G(d) and B3LYP/DZVP levels for [CuL(2)](+) and [CuL(2)][PF(6)] (L = 6,6'-dimethyl-2,2' bipyridine-4,4'-dimethylformate), both in the gas phase and in methyl cyanide (MeCN) solution. The vertical excitation energies were calculated within the framework of the time-dependent DFT (TD-DFT) approach, whereas the solvent effects were taken into account using the polarizable continuum model (C-PCM). Our results show that the five highest occupied molecular orbitals (HOMOs) are composed of a set of distorted degenerate Cu 3d orbitals, whereas the four lowest unoccupied molecular orbitals (LUMOs) are the bipyridine ligand pi*(C horizontal lineN) orbitals. The spectra in the range of 400-600 nm were found to originate from metal-to-ligand charge-transfer (MLCT) transitions, whereas the spectra in the range of 350-400 nm are excitations mainly from the metal Cu 3d orbitals to the carboxyl pi* orbitals. The solvent effects lead to changes in both the geometries and the absorption spectra. The results of this work suggest that copper-based complexes might be effective sensitizers for next-generation dye sensitized solar cells. PMID- 20000484 TI - Reticulon RTN1-C(CT) peptide: a potential nuclease and inhibitor of histone deacetylase enzymes. AB - RTN1-C protein is a membrane protein localized in the ER and expressed in the nervous system, and its biological role is not completely clarified. Our previous studies have shown that the C-terminal region of RTN1-C, corresponding to the fragment from residues 186 to 208, was able to bind the nucleic acids and to interact with histone deacetylase (HDAC) enzymes. In the present work the properties of the synthetic RTN1-C(CT) peptide corresponding to this region were studied with relation to its ability to bind the metal ions in its N-terminal region. RTN1-C(CT) peptide is characterized by the presence of high-affinity copper and nickel ion sites. The nuclease activity of the metal-peptide complex was observed due to the presence of an ATCUN-binding motif. Moreover, the effect of the Cu/Ni-RTN1-C(CT) complexes on the HDAC activity was investigated. The histone deacetylase inhibitors are a new class of antineoplastic agents currently being evaluated in clinical trials. Our data show that the acetylated form of the metal-peptide complex is able to inhibit the HDAC activity at micromolar concentrations. These results allow to propose the Cu/Ni-RTN1-C(CT) complexes as models for the design of antitumor agents. PMID- 20000485 TI - Cyanide lability and linkage isomerism of hexacyanochromate(III) induced by the Co(II) ion. AB - Reactions between the [M(III)(CN)(6)](3-) (M = Fe and Co) anions and the mononuclear complex [Co(II)(dppe)(2)(H(2)O)][BF(4)](2) result in the formation of two isostructural trinuclear clusters {[Co(II)(dppe)(2)](2)[M(III)(CN)(6)]}(BF(4)). Surprisingly, reactions of [Co(dppe)(2)(H(2)O)](BF(4))(2) and [Co(triphos)(CH(3)CN)(2)](BF(4))(2) with [Cr(CN)(6)](3-) yield the mononuclear complexes [Co(dppe)(2)(CN)](BF(4)) and [Co(triphos)(CN)(2)], respectively. In the former case, an unusual pentanuclear intermediate complex {[Co(II)(3)(dppe)(4)(MeCN)][Cr(III)(CN)(6)](2)} was isolated. The reaction was probed by solution IR spectroscopy, which revealed a gradual conversion of the nu(C[triple bond]N) stretches of the starting materials to those of the CN-bridged intermediate and eventually to the single nu(C[triple bond]N) stretch of the final mononuclear product. The loss of carbon-bound CN(-) ligands from [Cr(CN)(6)](3-) occurs on a sufficiently slow time-scale for observation of varying degrees of cyanide linkage isomerism in the trigonal bipyramidal complex {[Co(tmphen)(2)](3)[Cr(CN)(6)](2)}; the study was aided by the use of different Co(II) starting materials. Results obtained by a combination of X-ray crystallography, infrared spectroscopy, and magnetometry provide unequivocal evidence that the presence of certain Lewis acids (e.g., Co(II) in this work and Fe(II) ions and BPh(3) in previously reported studies) promote the process of cyanide linkage isomerism, which, in the case of Co(II) species, leads to facile labilization of cyanide ligands from the [Cr(CN)(6)](3-) anion. PMID- 20000486 TI - Ion-transfer voltammetric behavior of protein digests at liquid/liquid interfaces. AB - The development of new methods for the detection of proteins and peptides is of widespread importance. In this work, the electrochemical behavior of peptide mixtures resulting from proteolytic digestion of proteins was investigated at the polarized liquid|liquid interface (or the interface between two immiscible electrolyte solutions, ITIES). The influence of pepsin digestion on three proteins (hemoglobin, lysozyme, and cytochrome c) was studied, and it was revealed that resulting cyclic voltammograms of the three protein digests were different due to the unique peptide mixtures for a given protein. Differential pulse stripping voltammetry of protein digests enabled the detection of digested proteins at concentrations ranging between 0.55 and 4.22 microM. A limit of detection of 0.55 microM of the initial concentration of protein was achieved, demonstrating the analytical possibilities of such an electrochemical method. These results show that ion-transfer voltammetry offers the opportunity to study and develop label-free detection of peptides resulting from enzymatic digestions of proteins and may thus have a role in development of new proteomic technologies. PMID- 20000487 TI - Pd-catalyzed intramolecular oxyalkynylation of alkenes with hypervalent iodine. AB - The first example of intramolecular oxyalkynylation of nonactivated alkenes using oxidative Pd chemistry is reported. Both phenol and aromatic or aliphatic acid derivatives could be used under operator-friendly conditions (room temperature, technical solvents, under air). The discovery of the superiority of benziodoxolone-derived hypervalent iodine reagent 3d as an alkyne transfer reagent further expands the rapidly increasing utility of hypervalent iodine reagents in catalysis and is expected to have important implications for other similar processes. PMID- 20000488 TI - Aryl sulfoxides from allyl sulfoxides via [2,3]-sigmatropic rearrangement and domino Pd-catalyzed generation/arylation of sulfenate anions. AB - Allylic sulfoxides, via [2,3]-sigmatropic rearrangement and oxidative addition of the resulting allylic sulfenate esters to Pd(0), are found to be excellent precursors of sulfenate anions. This hitherto unknown reactivity is applied in a new Pd(0)-catalyzed domino sequence involving sulfenate anion generation followed by arylation to afford aryl sulfoxides. PMID- 20000489 TI - Bis-4-aminobenzamidines: versatile, fluorogenic A/T-selective dsDNA binders. AB - N(1),N(3)-Bis(4-amidinophenyl)propane-1,3-diamine (BAPPA, R = H) is a bisaminobenzamidine fluorogenic derivative that displays a large increase in its emission fluorescence when bound to the minor groove of specific A/T DNA sites (K(d) for an AATT site approximately 79 +/- 6 nM). Moreover, the structural characteristics of BAPPA allow the easy introduction of functional groups that protrude out of the DNA surface. PMID- 20000490 TI - An unusual pattern of CytR and CRP binding energetics at Escherichia coli cddP suggests a unique blend of class I and class II mediated activation. AB - Two transcription factors, CRP and CytR, mediate positive and negative control of nine cistrons involved in nucleoside catabolism and recycling in Escherichia coli. The ability of multiple transcription factors to combine in different ways to confer differential gene regulation is of significant interest in both prokaryotic and eukaryotic gene regulation. Analysis of cooperative interactions between CytR and CRP at the deoP2 and udpP promoters has implicated the importance of promoter architecture in controlling repression and induction. These studies have also identified competition between CytR and CRP as an additional contributor to differential regulation. The pattern and energetics of CytR and CRP interactions at the cdd promoter, the most strongly activated of the CytR-regulated promoters, have been delineated using DNase I footprinting. Surprisingly, CRP has greater affinity for the promoter proximal site at cddP, CRP1, than for the distal site, CRP2, in contrast to promoters studied previously. This difference is a major contributor to unusually high CRP-mediated activation of cddP. Additionally, while cytidine binding to CytR nearly eliminates the pairwise interactions between CytR and CRP bound at CRP1, it has little effect on pairwise cooperativity between CytR and CRP bound at CRP2 or as a consequence on the overall cooperativity of the three-protein complex in which CRP is bound to both sites. The effect of cytidine binding on cooperativity differs between the three promoters studied thus far. We propose that the different patterns of interaction reflect the spacing between CytR half-sites and the location of the CytR operator in relation to the two CRP sites. PMID- 20000491 TI - Iterative change. PMID- 20000492 TI - Salt-loving algae wipe out fish in Appalachian stream. PMID- 20000493 TI - Study reveals new mercury risks for fish and birds. PMID- 20000494 TI - Land use as climate change mitigation. PMID- 20000495 TI - Wind deployment in the United States: states, resources, policy, and discourse. AB - A transformation in the way the United States produces and uses energy is needed to achieve greenhouse gas reduction targets for climate change mitigation. Wind power is an important low-carbon technology and the most rapidly growing renewable energy technology in the U.S. Despite recent advances in wind deployment, significant state-by-state variation in wind power distribution cannot be explained solely by wind resource patterns nor by state policy. Other factors embedded within the state-level socio-political context also contribute to wind deployment patterns. We explore this socio-political context in four U.S. states by integrating multiple research methods. Through comparative state-level analysis of the energy system, energy policy, and public discourse as represented in the media, we examine variation in the context for wind deployment in Massachusetts, Minnesota, Montana, and Texas. Our results demonstrate that these states have different patterns of wind deployment, are engaged in different debates about wind power, and appear to frame the risks and benefits of wind power in different ways. This comparative assessment highlights the complex variation of the state-level socio-political context and contributes depth to our understanding of energy technology deployment processes, decision-making, and outcomes. PMID- 20000496 TI - Evaluation of small arms range soils for metal contamination and lead bioavailability. AB - Although small arms ranges are known to be contaminated with lead, the full extent of metal contamination has not been described, nor has the oral bioavailability of lead in these soils. In this work, soil samples from ranges with diverse geochemical backgrounds were sieved to <250 microm and analyzed for total metal content. Soils had consistently high levels of lead and copper, ranging from 4549 to 24 484 microg/g and 223 to 2936 microg/g, respectively, while arsenic, antimony, nickel, and zinc concentrations were 100-fold lower. For lead bioavailability measurements, two widely accepted methods were used: an in vivo juvenile swine relative bioavailability method measuring lead absorption from ingested soils relative to equivalent lead acetate concentrations and an in vitro bioaccessibility procedure which measured acid-extractable lead as a percent of total lead in the soil. For eight samples, the mean relative bioavailability and bioaccessibility of lead for the eight soils was about 100% (108 +/- 18% and 95 +/- 6%, respectively) showing good agreement between both methods. Risk assessment and/or remediation of small arms ranges should therefore assume high bioavailability of lead. PMID- 20000497 TI - Occurrence of perfluoroalkyl carboxylates and sulfonates in drinking water utilities and related waters from the United States. AB - The prevalence and persistence of perfluoroalkyl compounds (PFCs) in environmental and biological systems has been well documented, and a rising number of reports suggest that certain PFCs can result in adverse health effects in mammals. As traditional water sources become increasingly impacted by waste discharge and the demand for planned potable reuse grows, there is recent interest in determining PFC occurrence in drinking water supplies. Here we report monitoring results from drinking water treatment facility samples collected across the United States, and from associated surface, ground, and wastewater sources. Using automated solid phase extraction (SPE) and isotope-dilution liquid chromatography/tandem mass spectrometry (LC/MS-MS), samples were screened for perfluorohexanoic acid (PFHxA), perfluorohexanesulfonate (PFHxS), perfluorooctanoic acid (PFOA), perfluorooctanesulfonate (PFOS), perfluorononanoic acid (PFNA) perfluorodecanoic acid (PFDA), perfluoroundecanoic acid (PFUdA), and perfluorododecanoic acid (PFDoA). Method reporting limits (MRLs) were established at 1.0 ng/L for all monitored PFCs except PFOA, for which the MRL was set at 5.0 ng/L given elevated procedural and instrumental background levels. PFOS was the only investigated PFC detected in minimally impacted surface waters, with individual site averages of 2.0 ng/L and lower. Conversely, wastewater treatment plant (WWTP) effluents and other highly impacted waters had almost 100% detection frequency for all PFCs except PFUdA and PFDoA, which were not detected above MRL in any samples. Of the investigated PFCs, PFOA averaged the highest overall concentration at any site at 115 ng/L. Substantial impacts from treated wastewater generally caused increased summed PFC concentrations at downstream drinking water facilities, although levels and distribution suggest geographical variability. No discernible differences between influent and effluent PFC levels were observed for drinking water facilities. Removal of PFCs, however, was observed at an indirect potable reuse facility using microfiltration and reverse osmosis for wastewater treatment, in which case all PFC levels in effluents were below the MRL. PMID- 20000498 TI - Trends in concentrations and use of agricultural herbicides for Corn Belt rivers, 1996-2006. AB - Trends in the concentrations and agricultural use of four herbicides (atrazine, acetochlor, metolachlor, and alachlor) were evaluated for major rivers of the Corn Belt for two partially overlapping time periods: 1996-2002 and 2000-2006. Trends were analyzed for 11 sites on the mainstems and selected tributaries in the Ohio, Upper Mississippi, and Missouri River Basins. Concentration trends were determined using a parametric regression model designed for analyzing seasonal variability, flow-related variability, and trends in pesticide concentrations (SEAWAVE-Q). The SEAWAVE-Q model accounts for the effect of changing flow conditions in order to separate changes caused by hydrologic conditions from changes caused by other factors, such as pesticide use. Most of the trends in atrazine and acetochlor concentrations for both time periods were relatively small and nonsignificant, but metolachlor and alachlor were dominated by varying magnitudes of concentration downtrends. Overall, with trends expressed as a percent change per year, trends in herbicide concentrations were consistent with trends in agricultural use; 84 of 88 comparisons for different sites, herbicides, and time periods showed no significant difference between concentration trends and agricultural use trends. Results indicate that decreasing use appears to have been the primary cause for the concentration downtrends during 1996-2006 and that, while there is some evidence that nonuse management factors may have reduced concentrations in some rivers, reliably evaluating the influence of these factors on pesticides in large streams and rivers will require improved, basin specific information on both management practices and use over time. PMID- 20000499 TI - Ultrafine particles in indoor air of a school: possible role of secondary organic aerosols. AB - The aim of this work was to investigate ultrafine particles (<0.1 microm) in primary school classrooms, in relation to the classroom activities. The investigations were conducted in three classrooms during two measuring campaigns, which together encompassed a period of 60 days. Initial investigations showed that under the normal operating conditions of the school there were many occasions in all three classrooms where indoor particle concentrations increased significantly compared to outdoor levels. By far the highest increases in the classroom resulted from art activities (painting, gluing, and drawing), at times reaching over 1.4 x 10(5) particle cm(-3). The indoor particle concentrations exceeded outdoor concentrations by approximately 1 order of magnitude, with a count median diameter ranging from 20 to 50 nm. Significant increases also occurred during cleaning activities, when detergents were used. GC-MS analysis conducted on 4 samples randomly selected from about 30 different paints and glues, as well as the detergent used in the school, showed that d-limonene was one of the main organic compounds of the detergent, however, it was not detected in the samples of the paints and the glue. Controlled experiments showed that this monoterpene, emitted from the detergent, reacted with O(3) (at outdoor ambient concentrations ranging from 0.06 to 0.08 ppm) and formed secondary organic aerosols. Further investigations to identify other liquids that may be potential sources of the precursors of secondary organic aerosols were outside the scope of this project, however, it is expected that the problem identified by this study could be more widely spread, since most primary schools use liquid materials for art classes, and all schools use detergents for cleaning. Further studies are therefore recommended to better understand this phenomenon and also to minimize exposure of school children to ultrafine particles from these indoor sources. PMID- 20000500 TI - Site-specific profiles of estrogenic activity in agricultural areas of California's inland waters. AB - To evaluate the occurrence and sources of compounds capable of feminizing fish in agriculturally impacted waterways of the Central Valley of California, water samples were extracted and subjected to chemical analyses as well as in vitro and in vivo measurements of vitellogenin in juvenile rainbow trout (Oncorhynchus mykiss). Among the 16 sites sampled, 6 locations frequently exhibited elevated concentrations of estrogenic substances with 17beta-estradiol equivalents up to 242 ng/L in vitro and 12 microg/kg in vivo. The patterns of activity varied among sites, with two sites showing elevated activity only in vitro, two showing elevated activity only in vivo, and two showing elevated activity in both assays. Sequential elution of solid-phase extraction (SPE) disks followed by bioassay guided fractionation was used to characterize water samples from the two locations where activity was observed in both bioassays. The highest estrogenic activity was observed in the most nonpolar fractions (80-100% methanol eluent) from the Napa River, while most of the activity in the Sacramento River Delta eluted in the 60% methanol eluent. Quantitative analyses of SPE extracts and additional HPLC fractionation of the SPE extracts by GC-MS/MS and LC-MS/MS indicated concentrations of steroid hormones, alkylphenol polyethoxylates, and herbicides that were at least 1-3 orders of magnitude below bioassay 17beta estradiol equivalent calculations. Given the different patterns of activity and chemical properties of the estrogenic compounds, it appears that estrogenic activity in these agriculturally impacted surface waters is attributable to multiple compounds. Further investigation is needed to identify the compounds causing the estrogenic activity and to determine the potential impacts of these compounds on feral fish. PMID- 20000501 TI - Long-term measurements of NO(3) radical at a semiarid urban site: 1. Extreme concentration events and their oxidation capacity. AB - Nitrate radical (NO(3)), an important nighttime tropospheric oxidant, was measured continuously for two years (July 2005 to September 2007) in Jerusalem, a semiarid urban site, by long-path differential optical absorption spectroscopy (LP-DOAS). From this period, 21 days with the highest concentrations of nitrate radical (above 220 pptv) were selected for analysis. Joint measurements with the University of Heidelberg's LP-DOAS showed good agreement (r = 0.94). For all daytime measurements, NO(3) remained below the detection limit (8.5 pptv). The highest value recorded was more than 800 pptv (July 27, 2007), twice the maximum level reported previously. For this subset of measurements, mean maximum values for the extreme events were 345 pptv (SD = 135 pptv). Concentrations rose above detection limits at sunset, peaked between midnight and early morning, and returned to zero at sunrise. These elevated concentrations of NO(3) were a consequence of several factors, including an increase in ozone concentrations parallel to a substantial decrease in relative humidity during the night; Mean nighttime NO(2) levels above 10 ppbv, which prevented a deficiency in NO(3) precursors; Negligible NO levels during the night; and a substantial decrease in the loss processes, which led to a lower degradation frequency and allowed NO(3) lifetimes to build up to a maximum mean of 25 min. The results indicate that the major sink pathway for NO(3) was direct homogeneous gas phase reactions with VOC, and a smaller indirect pathway via hydrolysis of N(2)O(5). The Jerusalem measurements were used to estimate the oxidation potential of extreme NO(3) levels at an urban location. The 24 h average potential of NO(3), OH, and O(3) to oxidize hydrocarbons was evaluated for 30 separate VOCs. NO(3) was found to be responsible for approximately 70% of the oxidation of total VOCs and nearly 75% of the olefinic VOCs; which was more than twice the VOC oxidation potential of the OH radical. These results establish the NO(3) radical as an important atmospheric oxidant in Jerusalem. PMID- 20000502 TI - Ultrafiltration/nanofiltration for the tertiary treatment of leather industry effluents. AB - Biologically treated effluents from the leather industry pose severe problems for the environment due in part to both the inorganic charge and the high nitrogen content associated with the organic charge. Pressure-driven membrane processes, namely ultrafiltration/nanofiltration (UF/NF) technology, were investigated for their selective retention of the organics and permeation of the inorganic fraction. Permeation experiments were carried out with two model solutions representative of a treated tannery effluent. UF and NF of these model solutions were assessed in terms of both their inorganic/organic fractionation capability and their permeation productivity. The UF membranes with MWCOs ranging from 10,000 to 1000 Da yield retentate streams enriched in organic compounds and permeate streams enriched in salts. Despite their high capacity for pure water permeation, they displayed low permeation fluxes, as the result of concentration polarization and fouling phenomena. NF 200 and NF 270 membranes associated fractionation capability with high permeation rates. Furthermore, these membranes demonstrated the highest permeate fluxes -30 kg/h/m(2) and 16 kg/h/m(2) for different model solutions, at the transmembrane pressure of 8 bar. Although these membranes had lower hydraulic permeabilities relative to the other membranes tested, they exhibited the best characteristics in terms of minimization of colloidal fouling. PMID- 20000503 TI - Occurrence of polybrominated diphenyl ethers in air and precipitation of the Pearl River Delta, South China: annual washout ratios and depositional rates. AB - On the basis of a one-year (from October 2006 to September 2007) sampling campaign, 34 air samples and 23 bulk precipitation samples were collected in the Pearl River Delta (PRD) in southern China and analyzed for polybrominated diphenyl ethers (PBDEs). Fifteen tri- to deca-BDE congeners (sum of which is defined as Sigma15PBDE) were detected in more than 70% of the samples. In three urban-rural regions, Sigma15PBDE concentrations ranged from 77 to 372 pg/m3 in air (particulate + vapor) and 1.98 to 15.5 ng/L in rain (particle+dissolved) from Dongguan, from 195 to 1450 pg/m3 in air and 4.71 to 17.2 ng/L in rain from Shunde, and from 23.7 to 148 ng/L in rain from Guangzhou. Among the BDE congeners, BDE-209 was the predominant component. Linear correlations between the gas-particle partition coefficients (Kp) and the subcooled vapor pressures (P(O/L)) of individual BDE congeners were observed for both the wet and dry seasons, but the slopes (-0.572 to -0.525) of the fitted equations all substantially deviated from equilibrium condition (slope = -1). The total washout ratio by bulk rainfalls was determined to be 2 x 103 for tri-BDEs and 6 x 104 for BDE-209. The estimated annual dry and wet depositional rates were 6720 and 2460 kg/yr, respectively, for BDE-209, and 7270 and 2940 kg/yr for Sigma15PBDE in the PRD, indicating a dominant pathway for PBDEs input into the PRD soil and aquatic environments. PMID- 20000504 TI - Variations in stable isotope fractionation of Hg in food webs of Arctic lakes. AB - Biotic and abiotic fractionation of mercury (Hg) isotopes has recently been shown to occur in aquatic environments. We determined isotope ratios (IRs) of Hg in food webs (zooplankton, chironomids, Arctic char) and sediments of 10 Arctic lakes from four regions and investigated the extent of Hg isotope fractionation. Hg IRs were analyzed by multicollector inductively coupled plasma mass spectrometry (MC-ICP/MS). Hg mass independent fractionation (MIF; Delta(199)Hg) and mass dependent fractionation (MDF; delta(202)Hg) were calculated and compared among samples. IRs of Hg in sediment were characterized mainly by MDF and low MIF (Delta(199)Hg -0.37 to 0.74 per thousand). However, all biota showed evidence of MIF, most pronounced in zooplankton (Delta(199)Hg up to 3.40 per thousand) and char (Delta(199)Hg up to 4.87 per thousand). Zooplankton takes up highly fractionated MeHg directly from the water column, while benthic organisms are exposed to sedimentary Hg, which contains less fractionated Hg. As evidenced by delta(13)C measurements, benthic chironomids make up a large proportion of char diet, explaining in part why MIF(char) < MIF(zooplankton) in lakes, where both samples were measured. Hg IRs in char varied among regions, while char from lakes from each region showed similar degrees of MIF. A MIF-offset was derived representing the mean MIF difference between sediment and fish, and indicated that fish in two regions retain sediment signatures altered by a consistent offset. Due to its minimal lake-to-catchment area and very high water retention time ( approximately 330 years), the meteor impact crater lake (Pingualuk) reflects a "pure" atmospheric Hg signature, which is modified only by aqueous in lake processes. All other lakes are also affected by terrestrial Hg inputs and sediment processes. PMID- 20000505 TI - Light-initiated transformations of fullerenol in aqueous media. AB - We provide the first evidence that a fullerene derivative can be extensively mineralized under environmental conditions by direct photolysis. Dissolved inorganic carbon (DIC) was identified as a major photoproduct of fullerenol, a hydroxylated C(60) molecule and the ratio of moles DIC produced to moles of fullerenol reacted reached 28 or approximately 47% of complete mineralization on extensive irradiation by simulated solar radiation. The direct photoreaction kinetics of fullerenol in dilute aqueous solution can be described by pH dependent biexponential rate expressions. This photoreaction slowed by a factor of 2 in nitrogen-saturated water. The oxygen dependence is attributed to photoinduced electron or hydrogen atom transfer from fullerenol to oxygen to produce superoxide ions with a quantum yield of 6.2 x 10(-4). Fullerenol can photosensitize the production of singlet oxygen ((1)O(2)) in dilute aqueous solution with quantum yields ranging from 0.10 in acidic water to 0.05 in neutral and basic solution. However our results indicate that chemical reactions involving diffusive encounters between (1)O(2) or superoxide and fullerenol are too slow to significantly contribute to the fast component of fullerenol photoreaction in sunlight. The pH dependence of the direct and sensitized photoreactions is attributed to changes in intramolecular hemiketal formation in fullerenol. PMID- 20000506 TI - Single-walled carbon nanotubes exhibit limited transport in soil columns. AB - The increased production and commercial use of nanomaterials combined with a lack of regulation to govern their disposal may result in their introduction to soils and ultimately into groundwater systems. In this study, we investigated the transport behavior of carboxyl-functionalized single-walled carbon nanotubes (SWNTs) in columns packed with a natural soil. In general, SWNT deposition (filtration) rate increased with increasing solution ionic strength, with divalent cations (Ca(2+)) being more effective in increasing SWNT retention than monovalent cations (K(+)). However, SWNT deposition rate over a very wide range of monovalent and divalent cation concentrations (0.03 to 100 mM) was relatively high and changed only slightly above 0.3 mM KCl or 0.1 mM CaCl(2). In contrast, filtration of another type of engineered carbon-based nanomaterial, namely aqueous fullerene (C(60)) nanoparticles (radius of 51 nm), was more sensitive to solution ionic strength, displaying lower deposition rate and more effective transport in soil than SWNTs. These observations indicate that physical straining governs SWNT filtration and transport under all the solution chemistries investigated in the present study. It is proposed that SWNT shape and structure, particularly the very large aspect ratio and its highly bundled (aggregated) state in aqueous solutions, as well as the heterogeneity in soil particle size, porosity, and permeability, collectively contribute to straining in flow through soil media. Our results suggest that SWNTs of comparable properties to those used in the present study will not exhibit substantial transport and infiltration in soils because of effective retention by the soil matrix. PMID- 20000507 TI - Adsorption and desorption of oxytetracycline and carbamazepine by multiwalled carbon nanotubes. AB - We investigated the adsorption-desorption by multiwalled carbon nanotubes (MWCNTs) of two pharmaceuticals, oxytetracycline (OTC) and carbamazepine (CBZ). The pharmaceuticals demonstrated relatively fast sorption kinetics on MWCNTs. All adsorption isotherms were nonlinear and fit the Polanyi-Manes model (PMM). The single point adsorption coefficient (K) values for OTC were more than 1 order of magnitude higher than those for CBZ on corresponding MWCNTs. The adsorbed volume capacity (Q(0)) and K values of PMM showed a significant relationship with surface areas and the meso- and micropore volume of MWCNTs for both chemicals. Depending on the MWCNT outer diameter, 13.8-25.2% and 62.7-90.6% of initially adsorbed OTC and CBZ, respectively, were desorbed after 200 h. The rate of desorption of both OTC and CBZ depended upon pH and the quantity of initially adsorbed pharmaceuticals, as well as aggregation in the case of OTC. PMID- 20000508 TI - Speciation of technetium(IV) in bicarbonate media. AB - The technetium isotope (99)Tc is a major fission product from nuclear reactors. Ultimately it is disposed of as radioactive waste since it has few applications outside of scientific research. Geochemical modeling of the dissolution of nuclear waste and of the solubility and speciation of the dissolved radionuclides in groundwater is an important part of the Performance Assessment for the safety of nuclear waste repositories. It relies on the availability of a critically assessed thermodynamic database. The potential of the Tc(VII)/Tc(IV) redox couple is measured here under various chemical conditions to verify the stoichiometries of Tc complexes and determine their stabilities: (i) -log(10)[H(+)] in the range 7.0-10.0, for 0.3, 0.6, and 0.7 M [CO(3)](total); (ii) [CO(3)](total) in the range 0.01-0.6 M at -log(10)[H(+)] approximately 8.6; and (iii) [Tc(VII)]/[Tc(IV] ratios of (6.02 10(-5) M)/(10(-6) M) and (6.02 10(-5) M)/(6.02 10(-5) M) at log(10)[H(+)] = -9.1 and [CO(3)](total) = 1 M. Assuming that Tc(VII), TcO(4)(-) is the only species which exists under all the above chemical conditions, the potentiometric results can be interpreted by considering the presence of two hydroxide-carbonate monomeric complexes. The hydrolysis equilibrium between these two complexes is Tc(CO(3))(OH)(2) + H(2)O <--> Tc(CO(3))(OH)(3)(-) + H(+) with log(10)[H(+)](1/2) = 8.69 +/- 0.20, which is consistent with the -8.3 +/- 0.6 corresponding hydrolysis constant of the NEA TDB review. 733 +/- 44 mV/SHE and 575 +/- 60 mV/SHE are measured for the standard potentials of the TcO(4)( )/Tc(CO(3))(OH)(2), and for the TcO(4)(-)/Tc(CO(3))(OH)(3)(-) redox couples respectively. The corresponding formation constants from TcO(OH)(2) are log(10)K(1,2) = 19.8 +/- 0.5 and log(10)K(1,3) = 10.5 +/- 0.5, to be compared with the 19.3 +/- 0.3 and 11.0 +/- 0.6 values proposed by the NEA TDB review. Note that these values have been converted for the formation reactions described here, thus the given values are not those of the NEA TDB review. However, Tc(CO(3))(OH)(2) is predicted to dominate over a surprisingly large range of chemical conditions. The monomeric character of the Tc(IV) complexes is verified in this study. PMID- 20000509 TI - pH-Dependent sorption of acidic organic chemicals to soil organic matter. AB - Due to their increased polarity, many contemporary biologically active chemicals exhibit acid functions and may thus dissociate to their anionic conjugated base at pH values typically present in the environment. Despite its negative charge, soil organic matter (SOM) has been demonstrated to be the main sorbent in soils, even for the anionic species of organic acids. Nevertheless, few data exist that allow for a systematic interpretation of the sorption of organic acids into SOM. Therefore, in this study, the sorption of the neutral and anionic species of 32 diverse organic acids belonging to nine different chemical groups to SOM was investigated. Partition coefficients were determined from HPLC retention volumes on a column packed with peat, at three Ca(2+)-concentrations and over a pH range of 4.5-7.5. The influence of Ca(2+)-concentrations on anion sorption was small (factor 2 in the usual environmental Ca(2+)-concentration range) and independent of molecular structure. Generally, the organic carbon-water partition coefficients, K(oc), of both the neutral and anionic species increased with increasing molecular size and decreased with increasing polarity. At an environmentally relevant Ca(2+)-concentration of 10 mM, the investigated anions sorbed between a factor of 7-60 less than the corresponding neutral acid. This factor was more homogeneous within a group of structurally related compounds. These results indicate that while similar nonionic interactions seem to govern the partitioning of both the neutral and anionic species into SOM, the electrostatic interactions of the anionic species with SOM are a complex and currently not well understood function of the type of acidic functional group. The HPLC-based, flow through method presented in this study was shown to yield consistent results for a wide range of organic acids in a high-throughput manner. It should therefore prove highly useful in further investigating how different acidic functional groups affect anion sorption to SOM. PMID- 20000510 TI - Atmospheric emission of PCDD/Fs, PCBs, hexachlorobenzene, and pentachlorobenzene from the coking industry. AB - The coking process is considered to be a potential source of unintentionally produced persistent organic pollutants (UP-POPs). However, intensive studies on the emission of UP-POPs from the coking industry are still very scarce. Emission of polychlorinated dibenzo-p-dioxins/dibenzofurans (PCDD/Fs), dioxin-like polychlorinated biphenyls (dl-PCBs), hexachlorobenzene (HxCBz), and pentachlorobenzene (PeCBz) covered under the Stockholm Convention were investigated for the coking process in this study. Stack gases from some typical coke plants in China were collected and analyzed to estimate the emission of UP POPs from the coking industry. Emission factors of 28.9 ng WHO-TEQ tonne(-1) for PCDD/Fs, 1.7 ng WHO-TEQ tonne(-1) for dl-PCBs, 596 ng tonne(-1) for HxCBz, and 680 ng tonne(-1) for PeCBz were derived based on the investigated data. The annual emissions from the global coking industry were estimated to be 15.8 g WHO TEQ for PCDD/Fs, 0.93 g WHO-TEQ for dl-PCBs, 333 g for HxCBz, and 379 g for PeCBz, respectively (reference year 2007). According to the distribution of PCDD/Fs, we argued for the de novo synthesis to be the major pathway of PCDD/F formation. With regard to the characteristics of dl-PCBs, the most abundant congener was CB-118, and the most dominant contributor to the total WHO-TEQ of dl PCBs was CB-126. PMID- 20000511 TI - Application of quantum chemical approximations to environmental problems: prediction of water solubility for nitro compounds. AB - Water solubility values for 27 nitro compounds with experimentally measured values were computed using the conductor-like screening model for real solvent (COSMO-RS) based on the density functional theory and COSMO technique. We have found that the accuracy of the COSMO-RS approach for prediction of water solubility of liquid nitro compounds is impressively high (the errors are lower than 0.1 LU). However, for some solid nitro compounds, especially nitramines, there is sufficient disagreement between calculated and experimental values. In order to increase the accuracy of predictions the quantitative structure-property relationship (QSPR) part of the COSMO-RS approach has been modified. The solubility values calculated by the modified COSMO-RS method have shown much better agreement with the experimental values (the mean absolute errors are lower than 0.5 LU). Furthermore, this technique has been used for prediction of water solubility for an expanded set of 23 nitro compounds including nitroaromatic, nitramines, nitroanisoles, nitrogen rich compounds, and some their nitroso and amino derivatives with unknown experimental values. The solubility values predicted using the proposed computational technique could be useful for the determination of the environmental fate of military and industrial wastes and the development of remediation strategies for contaminated soils and waters. This predictive capability is especially important for unstable compounds and for compounds that have yet to be synthesized. PMID- 20000512 TI - Modeled environmental concentrations of engineered nanomaterials (TiO(2), ZnO, Ag, CNT, Fullerenes) for different regions. AB - Engineered nanomaterials (ENM) are already used in many products and consequently released into environmental compartments. In this study, we calculated predicted environmental concentrations (PEC) based on a probabilistic material flow analysis from a life-cycle perspective of ENM-containing products. We modeled nano-TiO(2), nano-ZnO, nano-Ag, carbon nanotubes (CNT), and fullerenes for the U.S., Europe and Switzerland. The environmental concentrations were calculated as probabilistic density functions and were compared to data from ecotoxicological studies. The simulated modes (most frequent values) range from 0.003 ng L(-1) (fullerenes) to 21 ng L(-1) (nano-TiO(2)) for surface waters and from 4 ng L(-1) (fullerenes) to 4 microg L(-1) (nano-TiO(2)) for sewage treatment effluents. For Europe and the U.S., the annual increase of ENMs on sludge-treated soil ranges from 1 ng kg(-1) for fullerenes to 89 microg kg(-1) for nano-TiO(2). The results of this study indicate that risks to aquatic organisms may currently emanate from nano-Ag, nano-TiO(2), and nano-ZnO in sewage treatment effluents for all considered regions and for nano-Ag in surface waters. For the other environmental compartments for which ecotoxicological data were available, no risks to organisms are presently expected. PMID- 20000513 TI - Assessing persistence and long-range transport potential of current-use pesticides. AB - Despite the fact that current-use pesticides (CUP) have different chemical properties to first-generation organochlorine products, the long-term and long range environmental behavior of these chemicals is still unclear. Data for 45 active ingredients of CUPs were collected, most of which originate from the results of simulation tests submitted for authorization. According to the Stockholm Convention on persistent organic pollutants (POPs), two of the 45 CUPs exceed both screening level criteria for persistence and long-range transport potential (LRTP). Thirteen CUPs meet the persistence criterion and just one for LRTP. This classification is compared to the reference chemicals approach using overall persistence (P(ov)) and characteristic travel distance (CTD) calculated with a multimedia model. Although none of the 45 CUP have a CTD above the LRTP boundary line, three of them exceed the overall persistence criterion derived from legacy POPs for classification. Nineteen CUPs are transported over longer distances in water than in air. For such polar substances a LRTP boundary has yet to be defined. We recommend the multimedia model modeling approach to calculate P(ov) and LRTP as a second tier in persistence and LRTP assessment. PMID- 20000514 TI - Spatially resolved distribution models of POP concentrations in soil: a stochastic approach using regression trees. AB - Background concentrations of selected persistent organic pollutants (polychlorinated biphenyls, hexachlorobenzene, p,p'-DDT including metabolites) and polyaromatic hydrocarbons in soils of the Czech Republic were predicted in this study, and the main factors affecting their geographical distribution were identified. A database containing POP concentrations in 534 soil samples and the set of specific environmental predictors were used for development of a model based on regression trees. Selected predictors addressed specific conditions affecting a behavior of the individual groups of pollutants: a presence of primary and secondary sources, density of human settlement, geographical characteristics and climatic conditions, land use, land cover, and soil properties. The model explained a high portion of variability in relationship between the soil concentrations of selected organic pollutants and available predictors. A tree for hexachlorobenzene was the most successful with 76.2% of explained variability, followed by trees for polyaromatic hydrocarbons (71%), polychlorinated biphenyls (68.6%), and p,p'-DDT and metabolites (65.4%). The validation results confirmed that the model is stable, general and useful for prediction. The stochastic model applied in this study seems to be a promising tool capable of predicting the environmental distribution of organic pollutants. PMID- 20000515 TI - A map of European emissions and concentrations of PFOS and PFOA. AB - A spatially distributed data set of measured concentrations of perfluorooctansulfonate (PFOS) and perfluorooctanoate (PFOA) was used, together with climatological averages of river flow, to estimate their overall aqueous emissions from the European Continent. This estimate relies on the assumption that PFOA/S are conservative chemicals. PFOS correlates rather well with river basin population, and its emissions can be estimated by regression for nonmonitored catchments. As such, regression equations were derived in both linear and log-linear form, the latter explaining a much higher portion of variance. Unlike PFOS, PFOA discharges are strongly influenced by point emissions from industrial facilities; they only correlate with catchment population below a threshold of 0.5 tons per year, and point source industrial emissions cannot be neglected in the estimation of PFOA loads. The lumped loads of PFOA from diffuse and point sources are reasonably described by a single log-linear regression model as a function of population in the catchment, likely owing to the fact that fluoropolymer industries located in highly populated catchments are more frequent as well. Overall, by using the log-linear models derived in this paper, PFOS and PFOA discharges along the whole European river network to coastal areas in Europe have been estimated for the year 2007 to be in the order of 20 and 30 tons per year, respectively. PMID- 20000516 TI - Evaluating the economic viability of a material recovery system: the case of cathode ray tube glass. AB - This paper presents an analysis of the material recovery system for leaded glass from cathode ray tubes (CRTs) using a dynamic material flow analysis. In particular, the global mass flow of primary and secondary CRT glass and the theoretical capacities for using secondary CRT glass to make new CRT glass are analyzed. The global mass flow analysis indicates that the amount of new glass required is decreasing, but is much greater than the amount of secondary glass collected, which is increasing. The comparison of the ratio of secondary glass collected to the amount of new glass required from the mass flow analysis indicates that the material recovery system is sustainable for the foreseeable future. However, a prediction of the time at which the market for secondary glass will collapse due to excess capacity is not possible at the moment due to several sources of uncertainty. PMID- 20000517 TI - Estimated 2017 refrigerant emissions of 2,3,3,3-tetrafluoropropene (HFC-1234yf) in the United States resulting from automobile air conditioning. AB - In response to recent regulations and concern over climate change, the global automotive community is evaluating alternatives to the current refrigerant used in automobile air conditioning units, 1,1,1,2-tetrafluoroethane, HFC-134a. One potential alternative is 2,3,3,3-tetrafluoropropene (HFC-1234yf, also known as HFO-1234yf). We have developed a spatially and temporally resolved inventory of likely future HFC refrigerant emissions from the U.S. vehicle fleet in 2017, considering regular, irregular, servicing, and end-of-life leakages. We estimate the annual leak rate emissions for each leakage category for a projected 2017 U.S. vehicle fleet by state, and spatially apportion these leaks to a 36 km square grid over the continental United States. This projected inventory is a necessary first step in analyzing for potential atmospheric and ecosystem effects, such as ozone and trifluoroacetic acid production, that might result from widespread replacement of HFC-134a with HFC-1234yf. PMID- 20000518 TI - Uranyl adsorption onto hydrous ferric oxide-A re-evaluation for the diffuse layer model database. AB - The diffuse layer model (DLM) database of Dzombak and Morel was developed to quantify the adsorption of dissolved species onto the hydrous ferric oxide (HFO) surface, and contained numerous surface complexation reactions, including surface complexation reactions for uranyl (UO(2)(+2)) consisting of Hfo_sOUO(2)(+) and Hfo_wOUO(2)(+). However, these constants were not based upon experimentally obtained data, but rather were derived from linear free energy relationships (LFER) using log K(MOH) values. When compared to experimental data, the LFER derived constants for uranyl were shown to overestimate adsorption by a factor of 10 in some cases. At least 14 uranyl HFO data sets have been previously published and were used to re-estimate constants by coupling the geochemical computer code PHREEQC with UCODE_2005, an automated parameter optimization program. Five uranyl bearing surface complexation reactions were initially evaluated; the constants were optimized by allowing UCODE to incrementally vary selected log K(x)(int) values until the best fit to the experimental data was obtained. Assumptions consistent with the original DLM were retained. Changes to the K(1)(int) and K(2)(int) constants, and addition of uranyl monocarbonate and uranyl dicarbonate surface complexes, will update and correct the uranyl sorption reactions in this widely used database. PMID- 20000519 TI - Characterizing attribute distributions in water sediments by geostatistical downscaling. AB - Information about attributes such as contaminant concentrations or hydraulic properties in benthic sediments is typically obtained in core sections of varying lengths, and only the average value is measured in each section. However, an estimate of the attribute distribution at a uniform spatial resolution is often required for site characterization and the design of appropriate risk-based remediation alternatives. Because attributes exhibit spatial autocorrelation, geostatistical methods have become an essential tool for estimating their spatial distribution. The purpose of this paper is to optimally infer the spatial distribution of sampled attributes at a uniform resolution from fluvial core sampling data, using a downscaling technique formulated as a geostatistical inverse problem. We compare geostatistical downscaling to the more traditional approach of point-to-point ordinary kriging for a hypothetical case study, and for total organic carbon observations from the Passaic River, New Jersey. Although frequently used to interpolate measurements, ordinary kriging is shown not to be able to estimate the spatial distribution of attributes accurately, because this approach assumes that data are sampled at a uniform resolution. Geostatistical downscaling, on the other hand, is shown to resolve this problem by explicitly accounting for the relationship between the known average measurements and the unknown fine-resolution attribute distribution to be estimated. PMID- 20000520 TI - Modeling the global fate and transport of perfluorooctane sulfonate (PFOS) and precursor compounds in relation to temporal trends in wildlife exposure. AB - A global-scale fate and transport model was applied to investigate the historic and future trends in ambient concentrations of perfluorooctane sulfonate (PFOS) and volatile perfluorooctane sulfonyl fluoride (POSF)-based precursor compounds in the environment. First, a global emission inventory for PFOS and its precursor compounds was estimated for the period 1957-2010. We used this inventory as input to a global-scale contaminant fate model and compared modeled concentrations with field data. The main focus of the simulations was to examine how modeled concentrations of PFOS and volatile precursor compounds respond to the major production phase-out that occurred in 2000-2002. Modeled concentrations of PFOS in surface ocean waters are generally within a factor of 5 of field data and are dominated by direct emissions of this substance. In contrast, modeled concentrations of the precursor compounds considered in this study are lower than measured concentrations both before and after the production phase-out. Modeled surface ocean water concentrations of PFOS in source regions decline slowly in response to the production phase-out while concentrations in remote regions continue to increase until 2030. In contrast, modeled concentrations of precursor compounds in both the atmosphere and surface ocean water compartment in all regions respond rapidly to the production phase-out (i.e., decline quickly to much lower levels). With respect to wildlife biomonitoring data, since precursor compounds are bioavailable and degrade to PFOS in vivo, it is at least plausible that declining trends in PFOS body burdens observed in some marine organisms are attributable to this exposure pathway. The continued increases in PFOS body burdens observed in marine organisms inhabiting other regions may reflect exposure primarily to PFOS itself, present in the environment due to production and use of this compound as well as degradation of precursor compounds. PMID- 20000521 TI - Density functional investigation of the water exchange reaction on the gibbsite surface. AB - The water exchange reactions on the gibbsite surface have been investigated by density functional calculations (B3LYP/6-31G(d) level) combining the supermolecular model and PCM model in this paper, and the water exchange rate constants on the gibbsite surface have also been predicted. In the proposed reaction pathways, the clusters Al6(OH)18(H2O)6(0) and Al6(OH)12(H2O)12(6+) are used as the models of gibbsite surface and protonated gibbsite surface respectively to examine the effect of protonation of gibbsite surface on the water exchange rate constants. The activation energy barriers DeltaE(s) not equal to (aq) for Al6(OH)18(H2O)6(0) and Al6(OH)12(H2O)12(6+) are 28.6 and 27.2 kJ*mol 1, respectively. The reaction energies DeltaE(s) (aq) for Al6(OH)18(H2O)6(0) and Al6(OH)12(H2O)12(6+) are 2.9 and 14.4 kJ mol-1, respectively, indicating that hexacoordinate aluminum in the gibbsite surface is more stable. The log k(TST) for Al6(OH)18(H2O)6(0) and Al6(OH)12(H2O)12(6+) are 6.5 and 7.5 respectively, and the log k(ex) calculated by the given transmission coefficient for Al6(OH)18 (H2O)6(0) and Al6(OH)12(H2O)12(6+) are 2.4 and 3.4 respectively, indicating that the protonation of gibbsite surface promotes the water exchange reaction of gibbsite surface and accelerates the dissolution rate of gibbsite. The relationship between the calculated free energy and experimental rate constants was explored, and according to this relationship, the log k(ex) for Al6(OH)18(H2O)6(0) and Al6(OH)12(H2O)12(6+) are 2.5 and 3.1 respectively, close to the corresponding values calculated by the given transmission coefficient. The water exchange rate constant of gibbsite surface is close to those of K-MAl(12)(M = Al, Ga, and Ge) polyoxocations, but deviates from that of Al(H2O)6(3+), implying that the same reactions with similar structure have similar water exchange rate constants. PMID- 20000522 TI - A picture of polar iodinated disinfection byproducts in drinking water by (UPLC/)ESI-tqMS. AB - Iodinated disinfection byproducts (DBPs) are generally more toxic than their chlorinated and brominated analogues. Up to date, only a few iodinated DBPs in drinking water have been identified by gas chromatography/mass spectrometry. In this work, a method for fast selective detection of polar iodinated DBPs was developed using an electrospray ionization-triple quadrupole mass spectrometer (ESI-tqMS) by conducting precursor ion scan of iodide at m/z 126.9. With such a method, pictures of polar iodinated DBPs in chlorinated, chloraminated, and chlorine-ammonia treated water samples were achieved. By coupling state-of-the art ultra performance liquid chromatography (UPLC) to the ESI-tqMS, structures of 17 iodinated DBPs were tentatively proposed. The results fully demonstrate that, with respect to the DBP number/levels among the three disinfection processes, chloramination generally generated the most/highest iodinated DBPs, chlorination generally produced the fewest/lowest iodinated DBPs, and chlorine-ammonia sequential treatment formed iodinated DBPs lying in between; the numbers of iodinated DBPs in chloraminated Suwannee River Fulvic Acid (SRFA) and Humic Acid (SRHA) were nearly the same, but the levels of aliphatic iodinated DBPs were higher in the chloraminated SRFA while the levels of aromatic iodinated DBPs were higher in the chloraminated SRHA; a couple of nitrogenous iodinated DBPs were found in chloramination and chlorine-ammonia treatment. The ratio of total organic iodine levels in chlorine-ammonia sequential treatment and chloramination could be expressed as a function of the lag time of ammonia addition. PMID- 20000523 TI - Development and validation of chemical and biological analyses to determine the antiestrogenic potency of resin acids in paper mill effluents. AB - This study combined chemical analysis and bioassays of paper mill effluents and their components in order to determine their antiestrogenic activity. The bioassay comprised a yeast two-hybrid assay incorporating the estrogen receptor alpha (hERalpha) and an hERalpha competitive enzyme-linked immunosorbent assay (ELISA). Samples were fractionated by solid phase extraction (SPE) with a C18 disk and a Florisil cartridge to obtain four fractions. The final fraction, eluted with methanol from the Florisil cartridge after pre-extraction by the C18 disk, was the most active in the two-hybrid assay, and its antiestrogenic potency, expressed as the equivalent concentration to 4-hydroxytamoxifen (OHT), was 277 nM. Seven resin acids had antiestrogenic activity in the active fraction as determined by gas chromatography/mass spectrometry (GC/MS), and their concentration levels ranged from 0.11 to 12 microg/L. All the resin acids exhibited greater activity than OHT; their activity relative to OHT ranged from 2.8- to 4.0-fold in the yeast two-hybrid assay. Based on the chemical analysis data and relative potency of resin acids from the yeast two-hybrid assay, the contribution ratio of resin acids accounted for 72% of the observed antiestrogenic activity of the extract. Furthermore, no resin acid showed any affinity for the estrogen receptor in the ELISA. This study showed that analysis combining the SPE method and the yeast two-hybrid assay is likely to be effective for the comprehensive monitoring of resin acids in paper mill industrial discharge areas. PMID- 20000524 TI - Use of reflectance infrared spectroscopy for monitoring the metal content of the estuarine sediments of the Nerbioi-Ibaizabal River (Metropolitan Bilbao, Bay of Biscay, Basque Country). AB - Multivariate partial least-squares (PLS) calibration models have been developed for the spatial and seasonal simultaneous monitoring of 14 trace elements (Al, As, Cd, Co, Cr, Cu, Fe, Mg, Mn, Ni, Pb, Sn, V, and Zn) in sediments from 117 samples taken in the estuary of the Nerbioi-Ibaizabal River. Models were based on the chemometric treatment of diffuse reflectance near-infrared (NIR) and attenuated total reflectance (ATR) mid infrared (MIR) spectra, obtained from samples previously lyophilized and sieved with a particle size lower than 63 microm. Vibrational spectra were scanned in both, NIR and MIR regions. Developed PLS models, based on the interaction between trace elements and organic mater provide good screening tools for the prediction of trace elements concentration in sediments. PMID- 20000525 TI - Design and evaluation of a porous burner for the mitigation of anthropogenic methane emissions. AB - Methane constitutes 15% of total global anthropogenic greenhouse gas emissions. The mitigation of these emissions could have a significant near-term effect on slowing global warming, and recovering and burning the methane would allow a wasted energy resource to be exploited. The typically low and fluctuating energy content of the emission streams makes combustion difficult; however porous burners-an advanced combustion technology capable of burning low-calorific value fuels below the conventional flammability limit-are one possible mitigation solution. Here we discuss a pilot-scale porous burner designed for this purpose. The burner comprises a cylindrical combustion chamber filled with a porous bed of alumina saddles, combined with an arrangement of heat exchanger tubes for preheating the incoming emission stream. A computational fluid dynamics model was developed to aid in the design process. Results illustrating the burner's stable operating range and behavior are presented: stable ultralean combustion is demonstrated at natural gas concentrations as low as 2.3 vol%, with transient combustion at concentrations down to 1.1 vol%; the system is comparatively stable to perturbations in the operating conditions, and emissions of both carbon monoxide and unburned hydrocarbons are negligible. Based on this pilot-scale demonstration, porous burners show potential as a methane mitigation technology. PMID- 20000526 TI - Arsenic removal from Vietnamese groundwater using the arsenic-binding DNA aptamer. AB - Single-stranded DNA aptamers were generated from a random library to remove arsenic from Vietnamese groundwater. On the basis of significant arsenic contamination levels, three areas in Ha Nam province (Vinh Tru, Bo De, and Hoa Hau) and five areas near the Mekong River Delta (MR1-5) were selected as study areas. The aptamers were in vitro selected using an arsenic aptamer affinity column created by immobilizing arsenic on Affi-gel 10 resin. Quantitative analyses of the aptamer candidates Ars-1 to Ars-8 by surface plasmon resonance (SPR) revealed the Ars-3 aptamer to have the highest affinity to arsenate [(As(V)] and arsenite [As(III)] with a dissociation constant (K(d)) of 4.95 +/- 0.31 and 7.05 +/- 0.91 nM, respectively. The specific affinity interactions of the Ars-3 aptamer to arsenic were verified against other heavy metals. After obtaining successful removal results with a laboratory-prepared aqueous arsenic solution, Ars-3 was applied for removal of any arsenic present in the groundwater samples collected from the studied areas in Vietnam. Field results were also successful: various arsenic concentrations ranging from 28.1 to 739.2 microg/L were completely removed after 5 min of incubation with the arsenic-binding aptamer Ars-3. PMID- 20000527 TI - Role of gelling soluble and colloidal microbial products in membrane fouling. AB - The mechanism underlying gel layer formation on membrane surfaces from soluble and colloidal microbial products (SCMPs) produced under unfavorable operational conditions for membrane bioreactors (MBRs) has been investigated using supernatants from a bench-scale MBR. SCMPs can be grossly classified into gelling and nongelling SCMPs with the gelling fraction associated mostly with the polysaccharide content. The significant role played by multivalent metals in gel formation through metal-ligand complexation has been confirmed. Functional groups of the gelling SCMPs were determined by pH titration and zeta potential measurement as amine/phenolic sites (pK(a) 9.3 and 8.0), carboxylic sites (pK(a) 6.6, 4.9, and ca. 4.0), and phosphoric sites (pK(a) ca. 2.5). The carboxylic sites were more directly involved with multivalent cation complexation; however, the gelling propensity of the SCMP dispersion was minimally affected by pH change in the circum-neutral pH range, suggesting that the strong carboxylic sites were principally responsible for gel formation. The SCMPs demonstrated a high potential for gel formation given the high density of the strong carboxylic acid groups of about 0.44 mmol/g-TOC and a moderate calcium binding stability constant of about 4.9 x 10(3) M(-1). PMID- 20000528 TI - Promotion of Ag/H-BEA by Mn for lean NO reduction with propane at low temperature. AB - Effects of adding manganese to Ag/H-BEA for selective catalytic reduction of NO(x) with propane (C(3)H(8)-SCR) were investigated under a lean-burn condition. Mn addition significantly promotes the catalytic performance of Ag/H-BEA below 673 K. A Ag-Mn/H-BEA catalyst with equal metal weight of 3 wt % has the highest activity for C(3)H(8)-SCR among samples with a different bimetal loading. Manganese is mainly present in the 3+ and 4+ oxidation states in Ag-Mn/H-BEA catalysts. The major contributions of manganese suggested by the data presented in this paper are to catalyze the NO oxidation and stabilize silver in a dispersed Ag(+) state. The presence of silver enforces the transformation of a certain amount of Mn(3+) ions to Mn(4+) ions. The activity of Ag-Mn/H-BEA decreases slightly at low SO(2) concentrations (0-200 ppm) but decreases significantly at high SO(2) concentrations (400-800 ppm). In the presence of 10% H(2)O and 200 ppm SO(2), the inhibition of C(3)H(8)-SCR below 673 K is more significant than that at high temperature above 673 K. Ag-Mn/H-BEA is a promising catalyst for the removal of NO(x) from diesel engine exhaust. PMID- 20000529 TI - A novel combination of anaerobic bioleaching and electrokinetics for arsenic removal from mine tailing soil. AB - This study provides evidence that a hybrid method integrating anaerobic bioleaching and electrokinetics is superior to individual methods for arsenic (As) removal from mine tailing soil. Bioleaching was performed using static reactors in batch tests and flow conditions in column test, and each test was sequentially combined with electrokinetics. In the bioleaching, indigenous bacteria were stimulated by the injection of carbon sources into soil, leading to the mobilization of As with the concurrent release of Fe and Mn. Compared with the batch-type bioleaching process, the combined process showed enhanced removal efficiency in the equivalent time. Although the transport fluid bioleaching conditions were inadequate for As removal, despite long treatment duration, when followed by electrokinetics the combined process achieved 66.5% removal of As from the soil. The improvement of As removal after the combined process was not remarkable, compared with single electrokinetics, whereas a cost reduction of 26.4% was achieved by the reduced duration of electrokinetics. The As removal performance of electrokinetics was significantly dependent on the chemical species of As converted via microbial metal reduction in the anaerobic bioleaching. The synergistic effect of the combined process holds the promise of significant time and cost savings in As remediation. PMID- 20000530 TI - Effects of water management on cadmium and arsenic accumulation and dimethylarsinic acid concentrations in Japanese rice. AB - Rice consumption is a major source of cadmium and arsenic for the population of Asia. We investigated the effects of water management in rice paddy on levels of cadmium and arsenic in Japanese rice grains. Flooding increased arsenic concentrations in rice grains, whereas aerobic treatment increased the concentration of cadmium. Flooding for 3 weeks before and after heading was most effective in reducing grain cadmium concentrations, but this treatment increased the arsenic concentration considerably, whereas aerobic treatment during the same period was effective in reducing arsenic concentrations but increased the cadmium concentration markedly. Flooding treatment after heading was found to be more effective than flooding treatment before heading in reducing rice grain cadmium without a concomitant increase in total arsenic levels, although it increased inorganic arsenic levels. Concentrations of dimethylarsinic acid (DMA) in grain were very low under aerobic conditions but increased under flooded conditions. DMA accounted for 3-52% of the total arsenic concentration in grain grown in soil with a lower arsenic concentration and 10-80% in soil with a higher arsenic concentration. A possible explanation for the accumulation of DMA in rice grains is that DMA translocates from shoots/roots to the grains more readily than does inorganic arsenic. PMID- 20000531 TI - Surface complexation modeling of U(VI) adsorption by aquifer sediments from a former mill tailings site at Rifle, Colorado. AB - A study of U(VI) adsorption by aquifer sediment samples from a former uranium mill tailings site at Rifle, Colorado, was conducted under oxic conditions as a function of pH, U(VI), Ca, and dissolved carbonate concentration. Batch adsorption experiments were performed using <2 mm size sediment fractions, a sand sized fraction, and artificial groundwater solutions prepared to simulate the field groundwater composition. To encompass the geochemical conditions of the alluvial aquifer at the site, the experimental conditions ranged from 6.8 x 10( 8) to 10(-5) M in [U(VI)](tot), 7.2 to 8.0 in pH, 3.0 x 10(-3) to 6.0 x 10(-3) M in [Ca(2+)], and 0.05 to 2.6% in partial pressure of carbon dioxide. Surface area normalized U(VI) adsorption K(d) values for the sand and <2 mm sediment fraction were similar, suggesting a similar reactive surface coating on both fractions. A two-site two-reaction, nonelectrostatic generalized composite surface complexation model was developed and successfully simulated the U(VI) adsorption data. The model successfully predicted U(VI) adsorption observed from a multilevel sampling well installed at the site. A comparison of the model with the one developed previously for a uranium mill tailings site at Naturita, Colorado, indicated that possible calcite nonequilibrium of dissolved calcium concentration should be evaluated. The modeling results also illustrate the importance of the range of data used in deriving the best fit model parameters. PMID- 20000532 TI - Monochloramination of resorcinol: mechanism and kinetic modeling. AB - The kinetics of monochloramination of resorcinol, 4-chlororesorcinol, and 4,6 dichlororesorcinol have been investigated over the pH range of 5-12, at 23 +/- 2 degrees C. Monochloramine solutions were prepared with ammonia-to-chlorine ratios (N/Cl) ranging from 1.08 to 31 mol/mol. Under conditions that minimize free chlorine reactions (N/Cl > 2 mol/mol), the apparent second-order rate constants of monochloramination of resorcinol compounds show a maximum at pH values between 8.6 and 10.2. The intrinsic second-order rate constants for the reaction of monochloramine with the acid-base forms of the dihydroxybenzenes (Ar(OH)(2), Ar(OH)O(-), and Ar(O(-))(2)) were calculated from the apparent second-order rate constants. The stoichiometric coefficients for the formation of 4 chlororesorcinol by monochloramination of resorcinol and 4,6-dichlororesorcinol by monochloramination of 4-chlororesorcinol were found to be equal to 0.66 +/- 0.05 and 0.25 +/- 0.02 mol/mol, respectively at pH 8.6. A kinetic model that incorporates reactions of free chlorine and monochloramine with the different acid-base forms of resorcinol compounds simulated well the initial rates of degradation of resorcinol compounds and was useful to evaluate the contribution of free chlorine reactions to the overall rates of degradation of resorcinol at low N/Cl ratios. PMID- 20000533 TI - Comparison of toluene removal in air at atmospheric conditions by different corona discharges. AB - Different types of corona discharges, produced by DC of either polarity (+/-DC) and positive pulsed (+pulsed) high voltages, were applied to the removal of toluene via oxidation in air at room temperature and atmospheric pressure. Mechanistic insight was obtained through comparison of the three different corona regimes with regard to process efficiency, products, response to the presence of humidity and, for DC coronas, current/voltage characteristics coupled with ion analysis. Process efficiency increases in the order +DC < -DC < +pulsed, with pulsed processing being remarkably efficient compared to recently reported data for related systems. With -DC, high toluene conversion and product selectivity were achieved, CO(2) and CO accounting for about 90% of all reacted carbon. Ion analysis, performed by APCI-MS (Atmospheric Pressure Chemical Ionization-Mass Spectrometry), provides a powerful rationale for interpreting current/voltage characteristics of DC coronas. All experimental findings are consistent with the proposal that in the case of +DC corona toluene oxidation is initiated by reactions with ions (O(2)(+*), H(3)O(+) and their hydrates, NO(+)) both in dry as well as in humid air. In contrast, with -DC no evidence is found for any significant reaction of toluene with negative ions. It is also concluded that in humid air OH radicals are involved in the initial stage of toluene oxidation induced both by -DC and +pulsed corona. PMID- 20000534 TI - Catalytic seawater flue gas desulfurization model. AB - A model of a seawater flue gas desulfurization process (SFGD) where oxidation of the absorbed SO(2) is catalyzed by activated carbon is presented. The modeled SFGD process is comprised of two main units, an absorption packed scrubber, where SO(2) absorption takes place, and an oxidation basin, where the absorbed SO(2) is catalytically oxidized to sulfate, a natural component of seawater. The model takes into account the complex physical-chemical features of the process, combining mass-transfer, kinetics and equilibrium equations, and considering the electrolyte nature of the liquid phase. The model was validated with data from a SFGD pilot plant and a sensitivity analysis was performed, showing its predictive capability. The model is a useful tool for designing industrial desulfurization units with seawater. PMID- 20000535 TI - N(2)O production during nitrogen removal via nitrite from domestic wastewater: main sources and control method. AB - Nitrite has been commonly recognized as an important factor causing N(2)O production, which weakened the advantages of nitrogen removal via nitrite. To reduce and control N(2)O production from wastewater treatment plants, both long term and batch tests were carried out to investigate main sources and pathways of N(2)O production during nitrogen removal via nitrite from real domestic wastewater. The obtained results showed that N(2)O production during nitrogen removal via nitrite was 1.5 times as much as that during nitrogen removal via nitrate. It was further demonstrated that ammonia oxidization were main source of N(2)O production during nitrogen removal from domestic wastewater; whereas, almost no N(2)O was produced during nitrite oxidization and anoxic denitrification. N(2)O production during nitrogen removal via nitrite decreased about 50% by applying the step-feed SBR, due to the effective control of nitrite and ammonia, the precursors of N(2)O production. Therefore, the step-feed system is recommended as an effective method to reduce N(2)O production during nitrogen removal via nitrite from domestic wastewater. PMID- 20000536 TI - Stimulative effects of ozone on a biofilter treating gaseous chlorobenzene. AB - Recalcitrant volatile organic compounds with low biodegradabilities pose challenges for biofiltration technologies. In this study, the effects and mechanism of adding ozone on the performance of a biofilter were investigated. A biofilter treating chlorobenzene was set up and operated continuously for 265 days under different inlet ozone concentrations. Results showed that ozone below 120 mg m(-3) could notably enhance the biofilter performance. The average chlorobenzene removal efficiency increased from 40 to 70% and then to 90% while the inlet ozone concentration rose from 0 to 40 mg m(-3) and 120 mg m(-3). Reducing ozone concentration resulted in a decrease in removal efficiency from 90 to 40%. Further analysis indicated that the thickness and extra-cellular polymer substance content of the biofilm were remarkably reduced while inlet ozone concentration was gradually increased. Meanwhile, the specific surface areas of the filter bed were found to increase from 784 to 820 and 880 m(2) m(-3). A respiratory quinone profile showed that the dominant quinone shifted from ubiquinone-8 to menaquinone-9(H(2)) after ozone was added. This indicated that some Gram-positive bacteria with thick cell wall became the dominant species under ozone compression. PMID- 20000537 TI - Bioaugmentation with engineered endophytic bacteria improves contaminant fate in phytoremediation. AB - Phytoremediation of volatile organic contaminants often proves not ideal because plants and their rhizosphere microbes only partially degrade these compounds. Consequently, plants undergo evapotranspiration that contaminates the ambient air and, thus, undermines the merits of phytoremediation. Under laboratory conditions, endophytic bacteria equipped with the appropriate degradation pathways can improve in planta degradation of volatile organic contaminants. However, several obstacles must be overcome before engineered endophytes will be successful in field-scale phytoremediation projects. Here we report the first in situ inoculation of poplar trees, growing on a TCE-contaminated site, with the TCE-degrading strain Pseudomonas putida W619-TCE. In situ bioaugmentation with strain W619-TCE reduced TCE evapotranspiration by 90% under field conditions. This encouraging result was achieved after the establishment and enrichment of P. putida W619-TCE as a poplar root endophyte and by further horizontal gene transfer of TCE metabolic activity to members of the poplar's endogenous endophytic population. Since P. putida W619-TCE was engineered via horizontal gene transfer, its deliberate release is not restricted under European genetically modified organisms (GMO) regulations. PMID- 20000538 TI - Sensitivity analysis of the pressure-based direct integrity test for membranes used in drinking water treatment. AB - We conducted a sensitivity analysis of the commonly employed pressure-based direct integrity test (DIT), the most sensitive test for defects in low-pressure hollow fiber (LPHF) microfiltration and ultrafiltration systems used in drinking water treatment. Incorporating uncertainty to assess the practice of DIT, we find the resolution in some tests may be insufficient to verify the presence of a barrier to oocysts of Cryptosporidium. Applying distributions and boundaries derived from literature and practice, we solved for the defect size resolution (DSR) using Monte Carlo and Probability Bounds Analysis for five commercial membrane designs. Surface tension was modeled using annual temperature profiles from three rivers. Contact angle measurement error and variability were derived from literature, respectively, as a standard deviation of 5.7 degrees and +/- 9.6 degrees median change due to natural organic matter (NOM) fouling. These measures of contact angle uncertainty and variability were combined in a normal distribution with the discrete values currently applied. Additionally we considered model uncertainty, applying the maximum bubble pressure method, an established method of surface tension measurement in liquids in which the maximum air pressure in a submerged capillary is developed after the contact angle becomes zero prior to bubble formation. Where the DSR exceeds 3 microm the test design is not compliant with applicable drinking water regulations. Implications include uncertain and variable log-removal values (LRV) as determined by DIT due to the possible emergence of defects large enough to allow oocysts to pass without detection by the DIT. Specifically, we found the DSR may exceed 3 microm and may be as large as 8 microm. With the variable contact angle model, all lower bound possibilities are compliant, whereas the upper bound is over 80% noncompliant for three of five commercial designs. Using the Maximum Bubble Pressure Method, the lower bounds in three designs start to exceed 3 microm for between 50 and 100% of the produced water, whereas the upper bounds of the DSR completely exceed 3 microm for four of five commercial designs examined. PMID- 20000539 TI - Hierarchically macro-/mesoporous Ti-Si oxides photonic crystal with highly efficient photocatalytic capability. AB - Hierarchically macro-/mesoporous Ti-Si oxides photonic crystal (i-Ti-Si PC) with highly efficient photocatalytic activity has been synthesized by combining colloidal crystal template and amphiphilic triblock copolymer. It was found that the thermal stability of mesoporous structures in the composite matrix were improved due to the introduction of silica acting as glue and linking anatase nanoparticles together, and the photocatalytic activity of the i-Ti-Si PCs was affected by the calcination conditions. The influences of photonic and structural effect of the i-Ti-Si PCs on photocatalytic activity were investigated. Photodegradation efficiency of the i-Ti-Si PCs was 2.1 times higher than that of TiO(2) photonic crystals (i-TiO(2) PCs) in the photodegradation of Rhodamine B (RB) dye as a result of higher surface area. When the energy of slow photon (SP) was optimized to the abosorption region of TiO(2), a maximum enhanced factor of 15.6 was achieved in comparison to nanocrystalline TiO(2) films (nc-TiO(2)), which originated from the synergetic effect of SP enhancement and high surface area. PMID- 20000540 TI - Life-cycle case study comparison of permeable reactive barrier versus pump-and treat remediation. AB - A permeable reactive barrier (PRB) is a passive remediation technology, which over decades of use, may reduce lifetime environmental impacts when compared with a conventional pump-and-treat system (PTS). Greater material production requirements to install PRBs may offset the expected reductions in operational phase impacts and the trade-offs can be investigated in a life-cycle assessment (LCA). The life-cycle environmental impacts of a zerovalent iron (ZVI) containing PRB with a funnel and gate configuration and a PTS were compared in a case study. Potential impacts of the model PRB are driven by the ZVI reactive medium and the energy usage during construction, while for the PTS they are driven by the operational energy demand. Medium longevity governed the magnitude of the potential PRB impacts and the extent to which it was optimal relative to the PTS. Even at conservatively low estimates of longevity, the PRB offers significant environmental advantages in impact categories of human health and ozone depletion. The minimum ZVI longevity for PRB benefit over the PTS system in all impact categories was 10 years. Suggested PRB design innovations to reduce environmental impacts include the development of alternative reactive media and construction methods. PMID- 20000541 TI - Identifying priority pollutant sources: apportioning air toxics risks using positive matrix factorization. AB - Hazardous air pollutants or air toxics are pollutants that are known or suspected to cause serious health effects. This paper presents a methodology to quantify source contributions to air toxics health risks. First, a linear, no-threshold risk model was used to identify gas-phase organic air toxics that contribute significantly to cancer risks. Next, Positive Matrix Factorization (PMF) was performed on high time-resolved measurements of these air toxics, and the additive cancer risks associated with each factor was determined. Finally, the PMF factors were linked to sources and source classes (mobile, nonmobile, secondary/background) using a combination of meteorological data and comparisons with published source profiles. The analysis was performed using data from three sites in Pittsburgh, Pennsylvania: a downtown site near a heavily traveled bus route, a residential site adjacent to a heavily industrialized area, and an urban background site. At all three sites emissions from nonmobile sources were the dominant contributors to the cancer risks from air toxics included in the PMF model, including benzene and other air toxics often associated with mobile source emissions. Emissions from both large industrial sources, such as coke works and chemical facilities, and smaller point sources, such as dry cleaners, contributed significantly to the cancer risks at all sites. This method can provide insight for decision makers to prioritize sources for risk reduction. PMID- 20000542 TI - Conditional sampling for source-oriented toxicological studies using a single particle mass spectrometer. AB - Current particulate matter regulations control the mass concentration of particles in the atmosphere regardless of composition, but some primary and/or secondary particulate matter components are no doubt more or less toxic than others. Testing direct emissions of pollutants from different sources neglects atmospheric transformations that may increase or decrease their toxicity. This work describes a system that conditionally samples particles from the atmosphere depending on the sources or source combinations that predominate at the sampling site at a given time. A single particle mass spectrometer (RSMS-II), operating in the 70-150 nm particle diameter range, continuously provides the chemical composition of individual particles. The mass spectra indicate which sources are currently affecting the site. Ten ChemVol samplers are each assigned one source or source combination, and the RSMS-II controls which one operates depending on the sources or source combinations observed. By running this system for weeks at a time, sufficient sample is collected by the ChemVols for comparative toxicological studies. This paper describes the instrument and algorithmic design, implementation, and first results from operating this system in Fresno, CA, during summer 2008 and winter 2009. PMID- 20000543 TI - Determination of Dechlorane Plus in serum from electronics dismantling workers in South China. AB - Previous studies have reported high serum concentrations of polybrominated diphenyl ethers in residents of an electronic waste (e-waste) dismantling site in Guiyu town, South China. In this study, serum levels of Dechlorane Plus (DP), a chlorinated flame retardant of growing concern, were measured in residents of this region, and compared to those of a matched cohort from a nearby region in Haojiang, where fishing is the dominant industry. DP was detected in all the serum samples, with concentrations ranging from 7.8 to 465 ng/g lipid (median: 42.6 ng/g) and 0.93 to 50.5 ng/g lipid (median: 13.7 ng/g) in Guiyu and Haojiang, respectively. The Pearson's correlation coefficient between DP and decabromodiphenyl ether showed a positive relationship in Guiyu (r = 0.931, p < 0.01), but no correlation in Haojiang (r = 0.321, p = 0.34). The average anti-DP fractional abundances (f(anti) ratio) were 0.58 +/- 0.11 and 0.64 +/- 0.05 for Guiyu and Haojiang, respectively (p < 0.01). Tentatively identified dechlorination product ([-Cl + H]) was found in 19/40 serum samples (11/20 for Guiyu, 8/20 for Haojiang). These results suggest that the Guiyu and Haojiang cohorts experienced different dominant exposure routes. It is also possible that DP could bioaccumulate in a stereoselective manner, and undergo dechlorination metabolism. To the best of our knowledge, this is the first study to report DP levels in human serum. Further studies are needed to confirm the results of this study. PMID- 20000544 TI - Avian digestive tract simulation to study the effect of grit geochemistry and food on Pb shot bioaccessibility. AB - Lead shot dissolution was investigated in a dynamic in vitro simulated avian gizzard-intestine system. The method allows simulated digestive fluid to pass (at intervals) from a gizzardlike environment to an intestine-based one, and then considers the dissolution of Pb shot (0-3 pellets) in the presence of differing grit geochemistries (siliceous and calcareous) and variable amounts of food (0-4 g of partially milled wheat seed). Dissolved Pb levels in simulated gizzards were consistently higher in the presence of siliceous, than with calcareous, grit. This was also seen in simulated intestines, except when less food was used (0-1 g), when Pb levels in solution were higher in calcareous systems. The Pb concentrations in gizzard and intestine solutions increased directly with the number of Pb shot used. In all treatments Pb levels in intestine liquids were lower than in gizzard liquids. Calcareous grit simulations maintained 2.5-34 times more Ca in solution than those that used siliceous grit. Dietary supplementation with calcareous grit may reduce Pb bioaccessibility of ingested Pb shot in birds by reducing gizzard acidity, by enhancing Pb precipitation (as Pb-carbonate), and by promoting higher dissolved Ca levels in the intestine, which may then compete with Pb for intestinal absorption. PMID- 20000545 TI - Assessment of four commonly employed in vitro arsenic bioaccessibility assays for predicting in vivo relative arsenic bioavailability in contaminated soils. AB - Currently, a number of in vitro methods are in use worldwide to assess arsenic (As) bioaccessibility in soils. However, a dearth of research has been undertaken to compare the efficacy of the in vitro methods for estimating in vivo relative As bioavailability. In this study, As bioaccessibility in contaminated soils (n = 12) was assessed using four in vitro assays (SBRC, IVG, PBET, DIN). In vitro results were compared to in vivo relative As bioavailability data (swine assay) to ascertain which methodologies best correlate with in vivo data. Arsenic bioaccessibility in contaminated soils varied depending on the in vitro method employed. For the SBRC and IVG methods, As bioaccessibility generally decreased when gastric-phase values were compared to the intestinal phase. In contrast, extending the PBET and DIN assays from the gastric to the intestinal phase resulted in an increase in As bioaccessibility for some soils tested. Comparison of in vitro and in vivo results demonstrated that the in vitro assay encompassing the SBRC gastric phase provided the best prediction of in vivo relative As bioavailability (R(2) = 0.75, Pearson correlation = 0.87). However, relative As bioavailability could also be predicted using gastric or intestinal phases of IVG, PBET, and DIN assays but with varying degrees of confidence (R(2) = 0.53 0.67, Pearson correlation = 0.73-0.82). PMID- 20000546 TI - Measurements of black and organic carbon emission factors for household coal combustion in China: implication for emission reduction. AB - Household coal combustion is considered as the greatest emission source for black carbon (BC) and an important source for organic carbon (OC) in China. However, measurements on BC and OC emission factors (EF(BC) and EF(OC)) are still scarce, which result in large uncertainties in emission estimates. In this study, a detailed data set of EF(BC) and EF(OC) for household coal burning was presented on the basis of 38 coal/stove combination experiments. These experiments included 13 coals with a wide coverage of geological maturity which were tested in honeycomb-coal-briquette and raw-coal-chunk forms in three typical coal stoves. Averaged values of EF(BC) are 0.004 and 0.007 g/kg for anthracite in briquette and chunk forms and 0.09 and 3.05 g/kg for bituminous coal, respectively; EF(OC) are 0.06 and 0.10 g/kg for anthracite and 3.74 and 5.50 g/kg for bituminous coal in both forms, respectively. Coal maturity was found to be the most important influencing factor relative to coal's burning forms and the stove's burning efficiency, and when medium-volatile bituminous coals (MVB) are excluded from use, averaged EF(BC) and EF(OC) for bituminous coal decrease by 50% and 30%, respectively. According to these EFs, China's BC and OC emissions from the household sector in 2000 were 94 and 244 gigagrams (Gg), respectively. Compared with previous BC emission estimates for this sector (e.g., 465 Gg by Ohara et al., Atmos. Chem. Phys. 2007, 7, 4419-4444), a dramatic decrease was observed and was mainly attributed to the update of EFs. As suggested by this study, if MVB is prohibited as household fuel together with further promotion of briquettes, BC and OC emissions in this sector will be reduced by 80% and 34%, respectively, and then carbonaceous emissions can be controlled to a large extent in China. PMID- 20000547 TI - Nanoparticle emissions from a heavy-duty engine running on alternative diesel fuels. AB - We have studied the effect of three different fuels (fossil diesel fuel (EN590); rapeseed methyl ester (RME); and synthetic gas-to-liquid (GTL)) on heavy-duty diesel engine emissions. Our main focus was on nanoparticle emissions of the engine. Our results show that the particle emissions from a modern diesel engine run with EN590, GTL, or RME consisted of two partly nonvolatile modes that were clearly separated in particle size. The concentration and geometric mean diameter of nonvolatile nucleation mode cores measured with RME were substantially greater than with the other fuels. The soot particle concentration and soot particle size were lowest with RME. With EN590 and GTL, a similar engine load dependence of the nonvolatile nucleation mode particle size and concentration imply a similar formation mechanism of the particles. For RME, the nonvolatile core particle size was larger and the concentration dependence on engine load was clearly different from that of EN590 and GTL. This indicates that the formation mechanism of the core particles is different for RME. This can be explained by differences in the fuel characteristics. PMID- 20000548 TI - Characteristics of diesel truck emission in China based on portable emissions measurement systems. AB - Seventy-five diesel vehicles were measured in China using a portable emissions measurement system (PEMS). Particular matter (PM) emission factors and gaseous emission factors for Euro 0 (E0), Euro 1 (E1), Euro 2 (E2), and Euro 3 (E3) trucks were obtained under highway, urban, and rural driving conditions. Vehicle emission regulations in China have successfully reduced carbon monoxide (CO), hydrocarbons (HC) and PM by 62, 56, and 72% on average. Most of the emission reductions were achieved when the control technology went from E0 to E1 in Xi'an, and E2 to E3 in Beijing, which resulted in PM reductions of 79% associated with highway driving and 60% associated with urban or rural driving. Emission levels of oxides of nitrogen (NO(X)) were not improved from previous emission control steps. Compared with Xi'an, the emission rate is lower in Beijing, which is strong evidence of the effectiveness of the present comprehensive emission control strategy in Beijing. Emissions were grouped into driving bins that corresponded to the energy demand placed on the vehicles. By using this binning approach, it was found that E3 trucks were successfully controlling the high emission rates in aggressive driving bins, which led to the low average emission for E3 trucks. PMID- 20000549 TI - Sulfide remediation by pulsed injection of nitrate into a low temperature Canadian heavy oil reservoir. AB - Sulfide formation by oil field sulfate-reducing bacteria (SRB) can be diminished by the injection of nitrate, stimulating the growth of nitrate-reducing bacteria (NRB). We monitored the field-wide injection of nitrate into a low temperature (approximately 30 degrees C) oil reservoir in western Canada by determining aqueous concentrations of sulfide, sulfate, nitrate, and nitrite, as well as the activities of NRB in water samples from 3 water plants, 2 injection wells, and 15 production wells over 2 years. The injection water had a low sulfate concentration (approximately 1 mM). Nitrate (2.4 mM, 150 ppm) was added at the water plants. Its subsequent distribution to the injection wells gave losses of 5 15% in the pipeline system, indicating that most was injected. Continuous nitrate injection lowered the total aqueous sulfide output of the production wells by 70% in the first five weeks, followed by recovery. Batchwise treatment of a limited section of the reservoir with high nitrate eliminated sulfide from one production well with nitrate breakthrough. Subsequent, field-wide treatment with week-long pulses of 14 mM nitrate gave breakthrough at an additional production well. However, this trend was reversed when injection with a constant dose of 2.4 mM (150 ppm) was resumed. The results are explained by assuming growth of SRB near the injection wellbore due to sulfate limitation. Injection of a constant nitrate dose inhibits these SRB initially. However, because of the constant, low temperature of the reservoir, SRB eventually grow back in a zone further removed from the injection wellbore. The resulting zonation (NRB closest to and SRB further away from the injection wellbore) can be broken by batch-wise increases in the concentration of injected nitrate, allowing it to re-enter the SRB dominated zone. PMID- 20000550 TI - Selecting anode-respiring bacteria based on anode potential: phylogenetic, electrochemical, and microscopic characterization. AB - Anode-respiring bacteria (ARB) are able to transfer electrons contained in organic substrates to a solid electrode. The selection of ARB should depend on the anode potential, which determines the amount of energy available for bacterial growth and maintenance. In our study, we investigated how anode potential affected the microbial diversity of the biofilm community. We used a microbial electrolysis cell (MEC) containing four graphite electrodes, each at a different anode potential (E(anode) = -0.15, -0.09, +0.02, and +0.37 V vs SHE). We used wastewater-activated sludge as inoculum, acetate as substrate, and continuous-flow operation. The two electrodes at the lowest potentials showed a faster biofilm growth and produced the highest current densities, reaching up to 10.3 A/m(2) at the saturation of an amperometric curve; the electrode at the highest potential produced a maximum of 0.6 A/m(2). At low anode potentials, clone libraries showed a strong selection (92-99% of total clones) of an ARB that is 97% similar to G. sulfurreducens. At the highest anode potential, the ARB community was diverse. Cyclic voltammograms performed on each electrode suggest that the ARB grown at the lowest potentials carried out extracellular electron transport exclusively by conducting electrons through the extracellular biofilm matrix. This is supported by scanning electron micrographs showing putative bacterial nanowires and copious EPS at the lowest potentials. Non-ARB and ARB using electron shuttles in the diverse community for the highest anode potential may have insulated the ARB using a solid conductive matrix from the anode. Continuous-flow operation and the selective pressure due to low anode potentials selected for G. sulfurreducens, which are known to consume acetate efficiently and use a solid conductive matrix for electron transport. PMID- 20000551 TI - A solar-powered microbial electrolysis cell with a platinum catalyst-free cathode to produce hydrogen. AB - This paper reports successful hydrogen evolution using a dye-sensitized solar cell (DSSC)-powered microbial electrolysis cell (MEC) without a Pt catalyst on the cathode, indicating a solution for the inherent drawbacks of conventional MECs, such as the need for an external bias and catalyst. DSSCs fabricated by assembling a ruthenium dye-loaded TiO(2) film and platinized FTO glass with an I( )/I(3)(-) redox couple were demonstrated as an alternative bias (V(oc) = 0.65 V). Pt-loaded (0.3 mg Pt/cm(2)) electrodes with a Pt/C nanopowder showed relatively faster hydrogen production than the Pt-free electrodes, particularly at lower voltages. However, once the applied photovoltage exceeded a certain level (0.7 V), platinum did not have any additional effect on hydrogen evolution in the solar-powered MECs: hydrogen conversion efficiency was almost comparable for either the plain (71.3-77.0%) or Pt-loaded carbon felt (79.3-82.0%) at >0.7 V. In particular, the carbon nanopowder-coated electrode without Pt showed significantly enhanced performance compared to the plain electrode, which indicates efficient electrohydrogenesis, even without Pt by enhancing the surface area. As the applied photovoltage was increased, anodic methanogenesis decreased gradually, resulting in increasing hydrogen yield. PMID- 20000553 TI - Retraction. Palladium-catalyzed annulation of 2-(1-alkynyl)biphenyls with aryl iodides to disubstituted methylidenefluorenes. PMID- 20000554 TI - Print-and-peel fabricated passive micromixers. AB - Advection driven mixing is essential for microfluidics and poses challenges to the design of microdevices. Force transducers or complex channel configurations provide means for, respectively, active or passive disrupting of laminar flows and for homogenizing the composing fluids. Print-and-peel (PAP) is a nonlithographic fabrication technique that involves direct printing of masters for molding polymer components of microdevices. PAP, hence, allows for facile and expedient preparation of microfluidic devices, without requiring access to specialized microfabrication facilities. We utilized PAP for fabrication of microfluidic devices capable of turning, expanding, and contracting microflows. We examined the mixing capabilities of these devices under flow conditions of small Reynolds numbers (0.2-20) and large Peclet numbers (260-26 000), under which advection is the dominant mode of mass transfer. We focused on mixing channels with arched shapes and examined the dependence of the mixing performance on the turns and the expansions along the direction of the microflows. Three dimensional expansion and contraction, along with an increase in the modes of twisting of the laminar currents, improved the quality of mixing. The simplicity in the described fabrication of the investigated passive micromixers makes PAP an attractive alternative for expedient device prototyping. PMID- 20000555 TI - Contact between submicrometer silica spheres. AB - Recently, the scope of the investigation of the deformation mechanism extended to the micrometer and submicrometer regimes. The sphere-substrate contact method was usually used because it is rather difficult to make two micrometer or submicrometer spheres contact each other precisely. Here, we used the sphere sphere contact method via a novel, simple process to investigate the deformation of spheres. The silica particle size ranges from 400 to 900 nm. Traditionally, the harder the particle, the smaller both the contact radius and the adhesion force. Therefore, it is widely accepted that silica particles should undergo elastic deformation, but we found that silica particles underwent plastic deformation rather than elastic deformation because of van der Waals interaction. The contact radii were observed by scanning electron microscopy (SEM). PMID- 20000556 TI - DFT calculations of EPR parameters for copper(II)-exchanged zeolites using cluster models. AB - The coordination environment of Cu(II) in hydrated copper-exchanged zeolites was explored through the use of density functional theory (DFT) calculations of EPR parameters. Extensive experimental EPR data are available in the literature for hydrated copper-exchanged zeolites. The copper complex in hydrated copper exchanged zeolites was previously proposed to be [Cu(H(2)O)(5)OH](+) based on empirical trends in tetragonal model complex EPR data. In this study, calculated EPR parameters for the previously proposed copper complex, [Cu(H(2)O)(5)OH](+), were compared to model complexes in which Cu(II) was coordinated to small silicate or aluminosilicate clusters as a first approximation of the impact of the zeolitic environment on the copper complex. Interpretation of the results suggests that Cu(II) is coordinated or closely associated with framework oxygen atoms within the zeolite structure. Additionally, it is proposed that the EPR parameters are dependent on the Si/Al ratio of the parent zeolite. PMID- 20000557 TI - Steady-state and time-resolved spectroscopy of 2,2'-bipyridine-3,3'-diol in solvents and cyclodextrins: polarity and nanoconfinement effects on tautomerization. AB - The ground- and excited-state tautomerization of the 2,2'-bipyridine-3,3'-diol molecule (BP(OH)(2)) was studied in different solvents and in confined nanocavities of cyclodextrins (CDs) using steady-state and lifetime spectroscopic measurements. In all solvents, a dizwitterion (DZ) tautomer is produced in the excited state after intramolecular double-proton transfer. This tautomer is stabilized in the ground state in water only and produces two unique absorption peaks in the region of 400-450 nm. The DZ tautomer fluoresces in the green and as the solvent polarity increases, the fluorescence peak is blue-shifted (498 nm in cyclohexane versus 462 nm in water), and the fluorescence lifetime gets shorter (3.10 ns in cyclohexane versus 0.65 ns in water). The results indicate the sensitivity of this tautomer to solvent polarity, particularly the solvent's hydrogen-bonding capability. In water, another photoinduced tautomerization mechanism takes place via a water network solvating each of the two hydrogen bonding centers of the molecule. The second tautomer is detected as a small shoulder in the blue side of the fluorescence peak and has a lifetime of 5.40 ns. Using BP(OH)(2) to probe the nanocavities of aqueous CDs reveals the degree of hydrophobicity of the cavities and the different mechanisms of probe encapsulation. As the cavity size decreases in the order gamma-CD to beta-CD to alpha-CD, the cavity is more hydrophobic, which is reflected in an intensity decrease of the absorbance of the DZ tautomer and a red shift in its fluorescence peak. The measured lifetimes show the same trend and reveal how the probe interacts with the CD moiety. In gamma-CD, the probe is located near the secondary rim of the CD annulus, whereas in alpha-CD, the probe is completely sequestered between two CDs, and the hydrophobicity is close to that observed in cyclohexane. In beta-CD and its derivatives, the spectral changes and the measured lifetimes indicate that the CD cavity gets more hydrophobic as a result of methyl substitution of the primary and secondary hydroxyls of the beta-CD rims. In the fully methylated 2,3,6-tri-O-methyl-beta-CD, the probe is exposed to water near the secondary rim due to the steric effect at the entrance rim that prevents the probe from full encapsulation. PMID- 20000552 TI - Organic chemistry and biology: chemical biology through the eyes of collaboration. AB - From a scientific perspective, efforts to understand biology including what constitutes health and disease has become a chemical problem. However, chemists and biologists "see" the problems of understanding biology from different perspectives, and this has retarded progress in solving the problems especially as they relate to health and disease. This suggests that close collaboration between chemists and biologists is not only necessary but essential for progress in both the biology and chemistry that will provide solutions to the global questions of biology. This perspective has directed my scientific efforts for the past 45 years, and in this overview I provide my perspective of how the applications of synthetic chemistry, structural design, and numerous other chemical principles have intersected in my collaborations with biologists to provide new tools, new science, and new insights that were only made possible and fruitful by these collaborations. PMID- 20000558 TI - Cloud droplet activation of amino acid aerosol particles. AB - In this work we investigated the ability of a series of amino acids to act as cloud condensation nuclei using a static thermal gradient diffusion type cloud condensation nucleus counter. Particles of pure dry l-glycine, glycyl-glycine, l serine, l-methionine, l-glutamic acid, l-aspartic acid, and l-tyrosine were studied as well as internally mixed dry particles containing ammonium sulfate and one or two of the following amino acids: l-methionine, l-aspartic acid, or l tyrosine. The amino acids ranged in water solubility from high (>100 g/L), intermediate (10-100 g/L), low (3-10 g/L), to very low (<3 g/L). With the exception of l-methionine and l-tyrosine, all the studied pure amino acid particles activated as though they were fully soluble, although Kohler theory modified to account for limited solubility suggests that the activation of the intermediate and low solubility amino acids l-serine, l-glutamic acid, and l aspartic acid should be limited by solubility. Activation of mixed particles containing at least 60% dry mass of l-tyrosine was limited by solubility, but the activation of the other investigated mixed particles behaved as if fully soluble. In general, the results show that particles containing amino acids at atmospherically relevant mixture ratios are good cloud condensation nuclei. PMID- 20000559 TI - Conformational analysis and vibrational circular dichroism of tris(ethylenediamine)ruthenium(II) complex: a theoretical study. AB - The conformational preferences and vibrational circular dichroism of tris(ethylenediamine)ruthenium complex in two main configurations (Lambda) and (Delta), have been performed using density functional theory. We find that for the free [Ru(en)(3)](2+) ion in the Delta-configuration, the conformational stability order is Delta(deltadeltadelta) > Delta(lambdadeltadelta) > Delta(lambdalambdadelta) > Delta(lambdalambdalambda) and that for the Lambda configuration it is Lambda(deltadeltadelta) < Lambda(lambdadeltadelta) < Lambda(lambdalambdadelta) < Lambda(lambdalambdalambda). The energy differences between the four conformers for both the configurations Delta and Lambda are relatively small, but the activation barriers for ring inversion from one conformation to another are significant, as compared to other such systems. We trace the origin of these results to the lower oxidation state of Ru and relatively larger Ru-N bond length. We have also studied the effect of counterions on the conformational stability for Ru(en)(3)Cl(2.) Our results indicate a reverse stability order for the associated complex, Ru(en)(3) Cl(2) and higher activation barriers for ring inversion as compared to the free complex ion Ru(en)(3)(2+). It is because of larger hydrogen bonding interactions between the three N-H bonds and the chloride ion in these two conformers as compared to other conformations, which is also evident from the VCD spectra of N-H stretching modes. We also investigate IR spectra for all conformations in Delta- and Lambda configurations and together with energetics and VCD spectra elucidate the spectroscopic characteristics of Ru(en)(3)(2+) complexes with and without the associated counterions. PMID- 20000560 TI - Tuning the excited-state dynamics of GFP-inspired imidazolone derivatives. AB - The excited-state dynamics of five derivatives of the GFP-chromophore, which differ by the position and nature of their substituents, has been investigated in solvents of various viscosity and polarity and in rigid media using femtosecond resolved spectroscopy. In polar solvents of low viscosity, like acetonitrile or methanol, the fluorescence decays of all compounds are multiexponential, with average lifetimes of the order of a few picoseconds, whereas in rigid matrices (polymer films and low temperature glasses), they are single exponential with lifetimes of the order of a few nanoseconds and fluorescence quantum yields close to unity. Global analysis of the fluorescence decays recorded at several wavelengths and of the transient absorption spectra reveals the presence of several excited-state populations with slightly different fluorescence and absorption spectra and with distinct lifetimes. These populations are attributed to the existence of multiple ground-state conformers. From the analysis of the dependence of the excited-state dynamics on the solvent and on the nature of the substituents, it follows that the nonradiative deactivation of all these excited chromophores involves an intramolecular coordinate with large amplitude motion. However, depending on the solvent and substituent, additional channels, namely, inter- and intramolecular hydrogen bond assisted nonradiative deactivation, are operative. This allows tuning of the excited-state lifetime of the chromophore. Finally, an ultrafast photoinduced intramolecular charge transfer is observed in polar solvents with one derivative bearing a dimethylaminophenyl substituent. PMID- 20000561 TI - Cooperative effect of solvent in the neutral hydration of ketenimine: an ab initio study using the hybrid cluster/continuum model. AB - The detailed reaction pathways for the hydration of ketenimine by water and water clusters containing up to five explicit water molecules (CH(2) =C=NH + (n + 1)H(2)O --> CH(3)CONH(2) + nH(2)O, n = 0-4) in the gas phase have been investigated by the MP2 method in conjunction with the 6-31+G* and 6-311++G** basis sets, and the effects of bulk solvent are taken into account according to the conductor-like polarizable continuum model (COSMO). In the hybrid cluster/continuum model, apart from one directly attacking water molecule, four explicit water molecules participating in the water-assisted hydrolysis of ketenimine are divided into two groups, one involving in the proton relay and the other near the nonreactive nitrogen or carbon atom. Two possible reaction channels, water addition across the C=C bond or across the C horizontal lineN bond of ketenimine, are discussed. Our results indicate that the kinetically favorable mechanism involves an eight-membered ring transition state structure formed by a proton transfer chain of three water molecules. Meanwhile, two additional cooperative water molecules near the nonreactive region assist the hydration by engaging in hydrogen bonding to the substrate; such an interaction is found to be important in the hydration of ketenimine and other cumulenes. The lowest rate-determining activation barriers of C=C and C=N addition are 98.9 and 95.1 kJ/mol, respectively, suggesting that the two channels are competitive when more water molecules take part in the hydration. COSMO solution models do not modify the calculated energy barriers in a significant way. PMID- 20000562 TI - Structural guidance of the photocycle of channelrhodopsin-2 by an interhelical hydrogen bond. AB - Channelrhodopsin-2 (ChR2) is a light-gated cation channel and a member of the family of retinylidene photoreceptors. Since the demonstration of light-induced depolarization of ChR2-expressing animal cell membranes, it was increasingly exploited for light triggering of action potentials. ChR2 conducts cations upon light absorption that embodies retinal isomerization as the primary reaction and a structurally unknown opening mechanism. It is evident from spectroscopic data that protonation reactions at the Schiff base are part of the photocycle, comparable to other microbial-type rhodopsins. However, the connection between the processes at the chromophore site and the channel's pore remained enigmatic. Here, we use slow mutants of ChR2 that we generated by disturbing a postulated hydrogen bond when mutating C128 in the transmembrane (TM) helix 3 and D156 in TM helix 4. The lifetime of the mutants' open state is increased more than 100 times. We investigated the spectral properties of the slow mutants. Whereas the deprotonation of the Schiff base (yielding P390) occurs on the same time scale as that of the wild type, reprotonation to P520 is retarded in the slow mutants and their photocycle is split, leading to the presence of two photointermediates, P390 and P520, in the open state. The photoreactions of P390 and P520 lead to a quenching of the current in electrophysiological measurements. We conclude that the putative hydrogen bond between C128 and D156 is an important structural determinant of the channel's closing reaction. Furthermore, we show that the D156A mutant is even more suitable for light control of excitable cells than C128A. PMID- 20000563 TI - Dielectric properties of N,N-dimethylformamide aqueous solutions in external electromagnetic fields by molecular dynamics simulation. AB - Nonequilibrium molecular dynamics (NMD) simulations of the N, N-dimethylformamide (DMF) aqueous solution have been performed in the canonical ensemble (NVT) in both the absence and presence of external electromagnetic (E/M) field, using the SPC/E model for water and the optimized potentials for liquid simulations-all atom (OPLS-AA) model for DMF. The electromagnetic wave propagates in the z-axis direction with a frequency of 10 GHz, and the intensity of the electromagnetic field in the range 0 to 3 x 10(9) V/m is assumed in the simulation. The results indicate that as the intensity of the electromagnetic field increased, the dipole alignment of the DMF aqueous solution molecules became more pronounced and the molecular polarizability gradually increased. In addition, the hydration number and the static dielectric constant of DMF aqueous solutions decreased as the intensity of the electromagnetic field increased. PMID- 20000564 TI - Impacts of competitive inhibition, parent compound formation and partitioning behavior on the removal of antibiotics in municipal wastewater treatment. AB - We present a process model that predicts the removal of the antibiotic micropollutants, sulfamethoxazole (SMX), tetracycline (TCY), and ciprofloxacin (CIP), in an activated sludge treatment system. A novel method was developed to solve the inverse problem of inferring process rate, sorption, and correction factor parameter values from batch experimental results obtained under aerobic and anoxic conditions. Instead of spiking the batch reactors with reference substances, measurements were made using the xenobiotic organic micropollutant content of preclarified municipal sewage. Parent compound formation and removal were observed, and the model developed using the simulation software West showed limited efficiency to describe the selected micropollutants profiles, when growth substrate removal occurs. The model structure was optimized by accounting for competitive inhibition by readily biodegradable substrates on the cometabolic micropollutant biotransformation processes. Our results suggest that, under anoxic conditions, hydrophobicity-independent mechanisms can significantly impact solid-liquid partitioning that our model takes into account by using the sorption coefficient as a lumped parameter. Forward dynamic simulations were carried out to evaluate the developed model and to confirm it for SMX using data obtained in a full-scale treatment plant. Evaluation of measured and simulation results suggest that, robust model prediction can be achieved by approximating the influent load of chemicals biodegrading via a given parent compound, e.g., human conjugates, as an antibiotic mass that is proportional to the parent compound load. PMID- 20000565 TI - The double [3 + 2] photocycloaddition reaction. AB - A remarkable double [3 + 2] photocycloaddition reaction that results in the formation of fenestrane 2 from aromatic acetal 1 is reported. During the formation of 2, four carbon-carbon bonds, five new rings, and seven new stereocenters are created in a one-pot process. The reaction occurs in a sequential manner from the linear meta photocycloadduct 3, while the angular meta photocycloadduct 4 undergoes an alternative fragmentation-translocation photoreaction to afford angular tricycle 6. PMID- 20000566 TI - Structural differences among alkali-soluble arabinoxylans from maize (Zea mays), rice (Oryza sativa), and wheat (Triticum aestivum) brans influence human fecal fermentation profiles. AB - Human fecal fermentation profiles of maize, rice, and wheat bran and their dietary fiber fractions released by alkaline-hydrogen peroxide treatment (principally arabinoxylan) were obtained with the aim of identifying and characterizing fractions associated with high production of short chain fatty acids and a linear fermentation profile for possible application as a slowly fermentable dietary fiber. The alkali-soluble fraction from maize bran resulted in the highest short chain fatty acid production among all samples tested, and was linear over the 24 h fermentation period. Size-exclusion chromatography and (1)H NMR suggested that higher molecular weight and uniquely substituted arabinose side chains may contribute to these properties. Monosaccharide disappearance data suggest that maize and rice bran arabinoxylans are fermented by a debranching mechanism, while wheat bran arabinoxylans likely contain large unsubstituted xylose regions that are fermented preferentially, followed by poor fermentation of the remaining, highly branched oligosaccharides. PMID- 20000567 TI - Screening for feeding deterrent and insect growth regulatory activity of triterpenic saponins from Diploknema butyracea and Sapindus mukorossi. AB - Antifeeding and insect growth regulatory effects of saponins and its hydrolyzed products from Diploknema butyracea and Sapindus mukorossi on the insect pest Spodoptera litura (F.) were investigated in the laboratory. D. butyracea saponins as well as their hydrolyzed prosapogenins were found to be better biologically active in controlling pests. A concentration of 1200 and 3400 mg L(-1) alkaline and acid hydrolyzed D. butyracea saponins exhibited significant antifeeding and toxic effects to third instar larvae when compared to the emulsified water as control. The n-BuOH extract after prep-HPLC separation provided two saponins from the D. butyracea saponin mixture: 3-O-[beta-D-glucopyarnosyl-beta-d glucopyranosyl]-16-alpha-hydroxyprotobassic acid-28-O-[ara-glc-xyl]-ara (MI-I) and 3-O-beta-D-glucopyranosyl-glucopyranosyl-glucopyranosyl-16-alpha hydroxyprotobassic acid-28-O-[ara-xyl-ara]-apiose (MI-III). The single saponin extracted from the S. mukorossi saponin mixture was 3-O-[beta-D-xyl(OAc).beta-D arabinopyranosyl.beta-D-rhamnopyranosyl] hederagenin-28-O-[beta-D-glc.beta-D glc.beta-D-rhamnopyranosyl] ester (SM-I). Five days after saponin treatment on larvae, the growth index (GI50) was reduced from 0.92% to 1520 ppm in alkaline hydrolyzed D. butyracea saponins. Upon hydrolysis, growth regulatory activity was improved in S. mukorossi saponin, whereas very little difference was found in antifeedant activity. Hydrophile-lipophile balance is important for the proper functioning of saponin/prosapogenin/sapogenin, which could be achieved by manipulating the sugar molecule in the triterpenic skeleton. PMID- 20000568 TI - Bidirectional electron transfer in molecular tetrads. AB - Two-color excitation of a molecular tetrad leads to directed electron transfer in opposite directions along the molecular axis according to which chromophore is illuminated and gives a 40,000-fold disparity in the lifetimes of the resultant charge-separated states. PMID- 20000569 TI - Divalent and oxabridged neonicotinoids constructed by dialdehydes and nitromethylene analogues of imidacloprid: design, synthesis, crystal structure, and insecticidal activities. AB - A series of divalent and oxabridged neonicotinoids were synthesized by reactions of nitromethylene analogues of imidacloprid and dialdehydes, and their structures were confirmed by (1)H NMR, (13)C NMR, high-resolution mass spectroscopy, and X ray diffraction analysis. The bioassays indicated that some of them were endowed with excellent insecticidal activities against cowpea aphid ( Aphis craccivora ), armyworm ( Pseudaletia separata Walker), and brown planthopper ( Nilaparvata lugens ). Divalent neonicotinoid 6 and oxabridged 8a had higher activities than imidacloprid against cowpea aphids and armyworm; furthermore, the activity of 8a was 40.4-fold higher than that of imidacloprid against imidacloprid-resistant brown planthopper. PMID- 20000570 TI - Bioactive compounds from sour orange inhibit colon cancer cell proliferation and induce cell cycle arrest. AB - Epidemiological studies suggest that dietary limonoids and phytosterols offer protection from certain types of cancers. Potential cancer preventive constituents of sour orange (Citrus aurantium L.) were isolated and identified from ethyl acetate extract. The structures of the compounds were identified by one-dimensional (1H or 13C) and two-dimensional (1H-H and 1H-13C) nuclear magnetic resonance experiments as limonexic acid and beta-sitosterol glucoside (SG). The identified compounds were tested for the potential inhibition of human colon cancer cell (HT-29) proliferation, apoptosis, and also noncancerous cells (COS-1). Cell proliferation, arrest of the cell cycle, and induction of apoptosis were assessed by MTT assay, flow cytometry, and nuclear staining methods, respectively. The MTT assay indicated that both compounds exhibited significant inhibition at various concentrations. These compounds did not show any toxic effects on noncancerous cells. These compounds caused 4-5-fold increases in the counts of G2/M stage cells at 50 microM, indicating a potential role in cell cycle arrest. These findings support the hypothesis that limonoids and phytosterols are effective apoptosis-promoting agents and incorporation of enriched fractions of these compounds in the diet may serve to prevent colon cancer. To the best of our knowledge, this is the first report of the isolation, identification, and cell proliferation assay for limonexic acid and SG from sour orange. PMID- 20000571 TI - Oxidation of polysaccharides by galactose oxidase. AB - Galactose oxidase was used as a catalyst to oxidize selectively the C-6 hydroxyls of terminal galactose to carbonyl groups. The polysaccharides studied included spruce galactoglucomannan, guar galactomannan, larch arabinogalactan, corn fiber arabinoxylan, and tamarind seed xyloglucan, with terminal galactose contents varying from 6% to 40%. A multienzyme system was used, with catalase and horseradish peroxidase to enhance the action of galactose oxidase. An analysis technique was developed for the quantification of the reactive aldehydes with GC MS, utilizing NaBD4 reduction and acidic methanolysis. The best oxidation degrees of terminal galactosyls were obtained with xyloglucan (85% of galactose) and spruce galactoglucomannan (65% of galactose). The highest oxidation degree based on total carbohydrates was achieved with guar gum (28%), which had the highest galactose content. The oxidation resulted in changes in the physicochemical properties of the polysaccharide solutions, and the changes observed varied between the polysaccharides. The clearest change was in tamarind xyloglucan, which formed a gel after the oxidation. After the oxidation, larger particles were present in the solution of spruce galactoglucomannan, but changes in its rheological properties were not observed. PMID- 20000572 TI - Systemic induction of monoterpene biosynthesis in Origanumxmajoricum by soil bacteria. AB - Italian oregano (Origanumxmajoricum) was subjected to root system inoculation with three species of plant growth-promoting rhizobacteria (PGPRs) (Pseudomonas fluorescens, Bacillus subtilis, Azospirillum brasilense), and essential oil (EO) content and plant growth were measured. Composition of monoterpenes, a major EO component, was analyzed qualitative and quantitatively by gas chromatography. Total EO yield for plants inoculated with P. fluorescens or A. brasilense was 3.57 and 3.41 microg/mg fresh weight, respectively, approximately 2.5-fold higher than controls, without change of quantitative oil composition. The major EO compounds, cis- and trans-sabinene hydrate, gamma-terpinene, carvacrol, and thymol, showed increased biosynthesis. Carvacrol was the only terpene showing significant increase of R% in plants inoculated with A. brasilense. Plant growth parameters (shoot and root fresh and dry weights, numbers of leaves and nodes) were evaluated. Shoot fresh weight was significantly increased by all three PGPR species, but only P. fluorescens and A. brasilense increased root dry weight. These two species have clear commercial potential for economic cultivation of O.xmajoricum. Knowledge of the factors affecting yield and accumulation of monoterpenes is essential for improving production of these economically important plant compounds. PMID- 20000573 TI - Quantification of iprodione in dry basil using silica gel supported titanium dioxide. AB - Iprodione is an agricultural fungicide that is difficult to detect in foods by HPLC because it coelutes with natural compounds in the food. We previously showed that food matrix could be degraded with titanium dioxide powder (TP). Here we describe an improved method for detection of iprodione using silica gel supported titanium dioxide (SGT). To synthesize SGT, titania-sol was mixed with diethanolamine, 2-propanol, and titanium tetraisopropoxide. After titania-sol was infiltrated into the silica gel (particle diameter 4 mm), the mixture was dried and then heated. Crude basil extract containing iprodione was mixed with SGT in a quartz vial, and the vial was irradiated with a UV light to selectively decompose the matrix interfering with the iprodione determination. In HPLC chromatograms of the treated solution, the interference peak decreased 35 times faster with SGT than with TP. When SGT (11 g) was added to the extract (20 mL) of dry basil (2 g), black light irradiation for 30 min was enough to quantify iprodione. The recovery rate of iprodione was 99.1%. Thus, the photocatalytic cleanup method using SGT is effective for analyzing residual iprodione in dry basil. PMID- 20000574 TI - Antioxidant protection of eicosapentaenoic acid and fish oil oxidation by polyphenolic-enriched apple skin extract. AB - Two apple skin extracts (ASE) prepared from "Northern Spy" cultivar were examined for their antioxidant properties to inhibit lipid oxidation in aqueous eicosapentaenoic acid (EPA) emulsions and bulk fish oil. The ASE were effective in reducing the oxidation induced by heat, UV light and peroxyl radical, when the extent of oxidation of the emulsions and bulk oil was measured by using the ferric thiocyanate test, the thiobarbituric acid reactive substances assay and Rancimat. On the basis of total phenolic concentration of extracts, removal of sugars and organic acids (ASE 2) from crude ethanol extract of apple skins (ASE 1) enhanced the antioxidant properties in both the emulsion and bulk fish oil systems. The average induction times of accelerated oxidation at 50 to 80 degrees C of fish oil incorporated with ASE 2 (400 microg/mL) was similar to that of alpha-tocopherol (400 microg/mL) and butylated hydroxytoluene (200 microg/mL). PMID- 20000575 TI - Combined effects of probiotic fermentation and high-pressure extraction on the antioxidant, antimicrobial, and antimutagenic activities of deodeok (Codonopsis lanceolata). AB - This study was designed to evaluate the combined effects of probiotic fermentation and high-pressure extraction (HPE) on the functional properties of Codonopsis lanceolata. The ground C. lanceolata samples were anaerobically fermented with Lactobacillus acidophilus ADH, Bifidobacterium longum B6, Lactobacillus rhamnosus GG, or Lactobacillus paracasei at 37 degrees C for 10 days and subjected to 500 MPa at 50 degrees C for 30 min. The extraction yields of C. lanceolata samples were noticeably increased to 29-32% by HPE. The B. longum-fermented C. lanceolata samples extracted by high pressure (BLF-HPE) exhibited the highest antimicrobial activity (MIC < 14 mg/mL) against Listeria monocytogenes, Staphylococcus aureus, Shigella boydii, and Salmonella typhimurium. The nonfermented C. lanceolata samples extracted with high pressure (NF-HPE) had the highest total phenolic content (13.3 mg of GAE/g). The lowest effective concentrations (EC(50) and EC(0.5)) were 4.55 and 1.76 mg/mL, respectively, for NF-HPE extracts, indicating its highest antioxidant activity. The BLF-HPE and L. rhamnosus-fermented C. lanceolata samples extracted by high pressure (LRF-HPE) exhibited the highest antimutagenic activities in S. typhimurium TA 100, which were 82 and 83% inhibition, respectively. The use of probiotic fermentation and HPE can produce more biologically active compounds in C. lanceolata than the conventional solvent extraction method. The results provide pharmaceutically useful information for improving biological properties and an approach to drug discovery. PMID- 20000576 TI - Synthesis and antimicrobial evaluation of amphiphilic neamine derivatives. AB - The aminoglycoside antibiotics bind to the 16S bacterial rRNA and disturb the protein synthesis. One to four hydroxyl functions of the small aminoglycoside neamine were capped with phenyl, naphthyl, pyridyl, or quinolyl rings. The 3',4'- (6), 3',6- (7a), and the 3',4',6- (10a) 2-naphthylmethylene derivatives appeared to be active against sensitive and resistant Staphylococcus aureus strains. 10a also showed marked antibacterial activities against Gram (-) bacteria, including strains expressing enzymes modifying aminoglycosides, efflux pumps, or rRNA methylases. 7a and 10a revealed a weak and aspecific binding to a model bacterial 16S rRNA. Moreover, as compared to neomycin B, 10a showed a lower ability to decrease (3)H leucine incorporation into proteins in Pseudomonas aeruginosa. All together, our results suggest that the 3',4',6-tri-2-naphthylmethylene neamine derivative 10a should act against Gram (-) bacteria through a mechanism different from inhibition of protein synthesis, probably by membrane destabilization. PMID- 20000577 TI - Rational design of 5-phenyl-3-isoxazolecarboxylic acid ethyl esters as growth inhibitors of Mycobacterium tuberculosis. a potent and selective series for further drug development. AB - New antituberculosis (anti-TB) drugs are urgently needed to shorten the 6-12 month treatment regimen and especially to battle drug-resistant Mycobacterium tuberculosis (Mtb) strains. In this study, we have continued our efforts to develop isoxazole-based anti-TB compounds by applying rational drug design approach. The biological activity and the structure-activity relationships (SAR) for a designed series of 5-phenyl-3-isoxazolecarboxylic acid ethyl ester derived anti-TB compounds were investigated. Several compounds were found to exhibit nanomolar activity against the replicating bacteria (R-TB) and low micromolar activity against the nonreplicating bacteria (NRP-TB). The series showed excellent selectivity toward Mtb, and in general, no cytotoxicity was observed in Vero cells (IC(50) > 128 muM). Notably, selected compounds also retained their activity against isoniazid (INH), rifampin (RMP), and streptomycin (SM) resistant Mtb strains. Hence, benzyloxy, benzylamino, and phenoxy derivatives of 5-phenyl-3 isoxazolecarboxylic acid ethyl esters represent a highly potent, selective, and versatile series of anti-TB compounds and as such present attractive lead compounds for further TB drug development. PMID- 20000578 TI - Dual-channel sensing of volatile organic compounds with semiconducting nanoparticles. AB - Extracting multidimensional information from an individual transducer simultaneously is a promising alternative sensing strategy to traditional sensors. Here, we proposed a novel dual channel sensing method with simultaneously recording conductivity change of sensing material and chemiluminescence emission during catalytic oxidation of volatile organic compounds on tin oxide nanoparticles. The orthogonal and complementary electrical and optical signals have been obtained for each compound, which have been applied to discriminate 20 volatile organic compounds using hierarchical cluster analysis (HCA). Unknown samples from three groups at concentrations of 0.2%, 0.6%, and 1.0% have been successfully classified using linear discriminant analysis (LDA) with accuracies of 98.3%, 96.7%, and 98.3%, respectively. This dual channel sensing mode is a complement of semiconducting type gas sensors and quite promising for the development of chemical sensor arrays with multimode transducing principles. PMID- 20000579 TI - Experimental determination of enthalpies of solution of tetraphenyl porphyrin (TPP) and some metal derivatives, in chloroform: interpretation of the solvation processes at a molecular level. AB - The enthalpies of solution, Delta(sol)H(m), for 5,10,15,20-tetraphenylporphine (CA registry number 917-23-7, TPP), 5,10,15,20-tetraphenylporphine of Co(II), Ni(II), Cu(II), and Zn(II) (to be written as CoTPP, NiTPP, CuTPP, and ZnTPP) in chloroform, were calorimetrically measured at T = 298 K in the concentration ranging from 3.5 x 10(-5) to 2.8 x 10(-4) mol.kg(-1). Through the linear extrapolation of the experimental data, corresponding values at infinite dilution were determined as: Delta(sol)H(m) (ZnTPP) = (55.5 +/- 0.2) kJ x mol(-1), Delta(sol)H(m) (CoTPP) = (36.9 +/- 0.2) kJ x mol(-1), Delta(sol)H(m) (TPP) = (25.7 +/- 0.6) kJ x mol(-1), Delta(sol)H(m) (NiTPP) = (15.6 +/- 0.1) kJ x mol( 1), and Delta(sol)H(m) (CuTPP) = (15.6 +/- 0.1) kJ x mol(-1). The enthalpies of solvation for the five compounds were also determined using the previously published values for the enthalpy of sublimation, as well as complementary data from the literature. The values obtained are as follows: Delta(solv)H(m)(TPP) = (158.3 +/- 2.1) kJ x mol(-1), Delta(solv)H(m)(CoTPP) = -(154.1 +/- 2.0) kJ x mol( 1), Delta(solv)H(m)(CuTPP) = -(149.4 +/- 5.0) kJ x mol(-1) Delta(solv)H(m)(NiTPP) = -(141.4 +/- 4.0) kJ x mol(-1), and Delta(solv)H(m)(ZnTPP) = -(140.5 +/- 3.0) kJ x mol(-1). The results are analyzed in relation to several molecular properties such as ionic radius, electronic spectra, and Connolly surface. An explanation of the observed trends for solvation enthalpies is proposed. PMID- 20000580 TI - Palladium-catalyzed tandem amination reaction for the synthesis of 4-quinolones. AB - An efficient palladium-catalyzed tandem amination approach was developed in one step to afford functionalized 4-quinolones in good to excellent yields from easily accessible o-haloaryl acetylenic ketones and primary amines. PMID- 20000581 TI - Theoretical study of the peripheral disulfide bridge substituent effects on the antioxidant properties of naphthyridine diol derivatives. AB - The effect of a peripheral disulfide bridge substituent on the phenolic O-H bond dissociation energy (BDE) and the ionization potential (IP) of naphthyridine diol has been studied by density functional theory (DFT) calculation. Compared with naphthalene diol, the substituent of a peripheral disulfide bridge group is very efficient in reducing the BDE, whereas the insertion of nitrogen atoms into the naphthalenic ring only slightly changes the BDE of O-H bond but dramatically enhances the IP. It is similar with the stereoelectronic effect of the heterocyclic ring for the well-known alpha-tocopherol antioxidant and leads to a highly delocalized spin distribution. With the incorporation of these two aspects, a potential antioxidant is expected to be more active and more stable than alpha-tocopherol. PMID- 20000582 TI - Fabrication and characterization of paper-based microfluidics prepared in nitrocellulose membrane by wax printing. AB - Paper-based microfluidics is a promising technology to develop a simple, low cost, portable, and disposable diagnostic platform for resource-limited settings. Here we report the fabrication of paper-based microfluidic devices in nitrocellulose membrane by wax printing for protein immobilization related applications. The fabrication process, which can be finished within 10 min, includes mainly printing and baking steps. Wax patterning will form hydrophobic regions in the membrane, which can be used to direct the flow path or separate reaction zones. The fabrication parameters like printing mode and baking time were optimized, and performances of the wax-patterned nitrocellulose membrane such as printing resolution, protein immobilization, and sample purification capabilities were also characterized in this report. We believe the wax-patterned nitrocellulose membrane will enhance the capabilities of paper microfluidic devices and bring new applications in this field. PMID- 20000583 TI - The effect of calcium ions and peptide ligands on the relative stabilities of the calmodulin dumbbell and compact structures. AB - A combination of ion mobility and mass spectrometry methods was used to characterize the molecular shape of the protein calmodulin (CaM) and its complexes with calcium and a number of peptide ligands. CaM, a calcium-binding protein composed of 148 amino acid residues, was found by X-ray crystallography to occur both in a globular shape and in the shape of an extended dumbbell. Here, it was found, as solutions of CaM and CaM complexes were sprayed into the solvent free environment of the mass spectrometer, that major structural features of the molecule and the stoichiometry of the units constituting a complex in solution were preserved in the desolvation process. Two types of CaM structures were observed in our experiments: a compact and an extended form of CaM with measured cross sections in near-perfect agreement with those calculated for the known globular and extended dumbbell X-ray geometries. Calcium-free solutions yielded predominantly an extended CaM conformation. Ca(n)(2+)-CaM complexes were observed in calcium-containing solutions, n = 0-4, with the population of the compact conformation increasing relative to the elongated conformation as n increases. For n = 4, a predominantly compact globular conformation was observed. Solutions containing the peptide CaMKII(290-309), the CaM target domain of the Ca(2+)/calmodulin-dependent protein kinase II (CaMKII) enzyme, yielded predominantly globular Ca(4)(2+)-CaM-CaMKII(290-309) complexes. Similar results were obtained with the 26-residue peptide melittin. For the 14-residue C-terminal melittin fragment, on the other hand, formation of both a 1:1 and a 1:2 CaM peptide complex was detected. On the basis of the entirety of our results, we conclude that the collapse of extended (dumbbell-like) CaM structures into more compact globular structures occurs upon specific binding of four calcium ions. Furthermore, this calcium-induced structural collapse of CaM appears to be a prerequisite for formation of a particularly stable CaM-peptide complex involving peptides long enough to be engaged in interactions with both lobes of CaM. PMID- 20000584 TI - Comparison of intrinsic stacking energies of ten unique dinucleotide steps in A RNA and B-DNA duplexes. Can we determine correct order of stability by quantum chemical calculations? AB - High level ab initio methods have been used to study stacking interactions in ten unique base pair steps both in A-RNA and in B-DNA duplexes. The protocol for selection of geometries based on molecular dynamics (MD) simulations is proposed, and its suitability is demonstrated by comparison with stacking in steps at fiber diffraction geometries. It is shown that fiber diffraction geometries are not sufficiently accurate for interaction energy calculations. In addition, the protocol for selection of geometries based on MD simulations allows for the evaluation of the variability of the intrinsic stacking energies along the MD trajectories. The uncertainty in stacking energies (difference between the most and least stable geometry) due to the dynamical nature of systems can be, in some cases, as large as 3.0 kcal x mol(-1), which is almost 50% of the actual sequence dependence of base stacking energies (the energy difference between the most and least stable sequences). Thus, assessing the relative magnitude of the gas phase stacking energy using a single geometry for each sequence is insufficient to obtain an unambiguous order of gas phase stacking energies in canonical double helices. Though the ordering of ten unique dinucleotide steps cannot be definitive, some general conclusions were drawn. The stacking energies of base pair steps in A-RNA are more evenly separated compared to B-DNA, and their ordering is less sensitive to the dynamics of the system compared to be B-DNA. The most stable step both in B-DNA and A-RNA is the GC/GC [corrected] step that is well separated from the second most stable step CG/CG. [corrected] Also the least stable step (the CC/GG step) is well separated from the rest of the structures. The calculations further show that B-DNA stacking is favorable only marginally (on average by 1.14 kcal x mol(-1) per base pair step) over A-RNA stacking, and this difference vanishes after subtracting the stabilizing van der Waals effect of the thymine 5-methyl group that is absent in RNA. Basically, no correlation between the sequence dependence of gas phase stacking energies and the sequence dependence of DeltaG degrees(37) free energies used in nearest neighbor models was found either for B-DNA or for A-RNA. This reflects the complexity of the balance of forces that are responsible for the sequence dependence of thermodynamics stability of nucleic acids, which masks the effect of the intrinsic interactions between the stacked base pairs. PMID- 20000585 TI - Quantitative multiplex detection of pathogen biomarkers on multichannel waveguides. AB - No single biomarker can accurately predict disease. An ideal biodetection technology should be capable of the quantitative, reproducible, and sensitive detection of a limited suite of such molecules. To this end, we have developed a multiplex biomarker assay for protective antigen and lethal factor of the Bacillus anthracis lethal toxin using semiconductor quantum dots as the fluorescence reporters on our waveguide-based biosensor platform. The platform is extendable to a wide array of biomarkers, facilitating rapid, quantitative, sensitive, and multiplex detection, better than achievable by conventional immunoassay. Our assay allows for the sensitive (limit of detection 1 pM each), specific (minimal nonspecific binding), and rapid (15 min) detection of these biomarkers in complex biological samples (e.g., serum). To address the issue of reproducibility in measurement and to increase our sample throughput, we have incorporated multichannel waveguides capable of simultaneous multiplex detection of biomarkers in three samples in quadruplicate. In this paper, we present the design, fabrication, and development of multichannel waveguides for the simultaneous detection of lethal factor and protective antigen in serum. Evaluation of the multichannel waveguide shows an excellent concordance with single-channel data and effective, simultaneous, and reproducible measurement of lethal toxins in three samples. PMID- 20000586 TI - Direct compound-specific stable chlorine isotope analysis of organic compounds with quadrupole GC/MS using standard isotope bracketing. AB - A method has been developed for the direct determination of the stable chlorine isotope composition (delta(37)Cl) of organochlorines that eliminates sample preparation, achieves precision comparable to earlier techniques while improving the sensitivity, and makes use of benchtop gas chromatography-quadrupole mass spectrometry instruments (GCqMS). The method is based on the use of multiple injections (n = 8-10) of the sample, bracketed by a molecularly identical isotopic standard with known delta(37)Cl, determined using off-line thermal ionization mass spectrometry (TIMS). Mass traces of two isotopologues differing by one chlorine isotope were used to calculate delta(37)Cl values. Optimization of mass spectrometry and peak integration parameters as well as method validation was achieved using tetrachloroethene (PCE), p,p'-dichlorodiphenyltrichloroethane (DDT), and pentachlorophenol (PCP), spanning a delta(37)Cl range of -5.5 to +3.2 per thousand vs SMOC. Injecting 1.6-1100 pmol resulted in standard deviations (1sigma) of 0.6-1.3 per thousand, and the delta(37)Cl results agreed with values independently measured with TIMS. The method was tested by determining the Rayleigh fractionation during evaporation of pure liquid PCE, resulting in a chlorine isotopic enrichment factor of epsilon(Cl) = -1.1 +/- 0.4 per thousand. Furthermore, position-specific delta(37)Cl analysis based on analysis of DDT mass fragments was evaluated. The GCqMS-delta(37)Cl method offers a simplified yet sensitive approach for compound-specific chlorine isotope analysis. PMID- 20000587 TI - Recipes for the selection of experimental protein conformations for virtual screening. AB - The use of multiple X-ray protein structures has been reported to be an efficient alternative for the representation of the binding pocket flexibility needed for accurate small molecules docking. However, the docking performance of the individual single conformations varies widely, and adding certain conformations to an ensemble is even counterproductive. Here we used a very large and diverse benchmark of 1068 X-ray protein conformations of 99 therapeutically relevant proteins, first, to compare the performance of the ensemble and single conformation docking and, second, to find the properties of the best-performing conformers that can be used to select a smaller set of conformers for ensemble docking. The conformer selection has been validated through retrospective virtual screening experiments aimed at separating known ligand binders from decoys. We found that the conformers cocrystallized with the largest ligands displayed high selectivity for binders, and when combined in ensembles they consistently provided better results than randomly chosen protein conformations. The use of ensembles encompassing between 3 and 5 experimental conformations consistently improved the docking accuracy and binders vs decoys separation. PMID- 20000588 TI - Metal-free intramolecular aziridination of alkenes using hypervalent iodine based sulfonyliminoiodanes. AB - Intramolecular aziridination of alkenyl sulfonyliminoiodanes occurs thermally in the absence of conventional metal catalysts such as Rh(II) and Cu(II). In rigid molecular systems, conversions are near quantitative. The scope of the nonmetal process is related to the conformational flexibility of the alkenyl sulfonyliminoiodane. A mechanism is proposed involving formal 2 + 2 cycloaddition of the RSO(2)N=IPh group to the double bond followed by reductive elimination of PhI to yield the sulfonylaziridine. Green chemistry aspects of the process are highlighted. PMID- 20000590 TI - Vibrational circular dichroism of Delta-SAPR-8-tetrakis[(+) heptafluorobutyrylcamphorato]lanthanide(III) complexes with an encapsulated alkali metal ion. AB - The vibrational circular dichroism (VCD) spectra for Delta-M[Ln((+)-hfbc)(4)] (M(I) = Na, K, Rb, Cs; Ln(III) = La, Eu; abbreviated as M-Ln) and Delta-Cs[Yb((+) hfbc)(4)] (Cs-Yb) were observed in CDCl(3) solutions. The VCD signs and/or intensities are mostly different from those of the Delta-tris(beta-diketonato) transition-metal complexes. The M-Ln complexes give the strong VCD peak(s) near 1550 cm(-1). These VCD patterns are different from each lanthanide(III) complex, but their intensities change with variation of the alkali metal ion. A positive negative couplet VCD near 1550 cm(-1) for the Delta-M-La complexes is observed similarly for the Delta-tris(beta-diketonato) complexes, confirming the assignment to the Delta absolute configuration based on the exciton CD. The density functional theory calculated VCD for the two peaks at 1650 and 1550 cm( 1) reproduces the relative intensity and signs of the observed ones for the Delta M-La(III) complexes. PMID- 20000589 TI - The uremic toxin 3-indoxyl sulfate is a potent endogenous agonist for the human aryl hydrocarbon receptor. AB - The aryl hydrocarbon receptor (AHR) is a ligand-activated transcription factor involved in the regulation of multiple cellular pathways, such as xenobiotic metabolism and Th17 cell differentiation. Identification of key physiologically relevant ligands that regulate AHR function remains to be accomplished. Screening of indole metabolites has identified indoxyl 3-sulfate (I3S) as a potent endogenous ligand that selectively activates the human AHR at nanomolar concentrations in primary human hepatocytes, regulating transcription of multiple genes, including CYP1A1, CYP1A2, CYP1B1, UGT1A1, UGT1A6, IL6, and SAA1. Furthermore, I3S exhibits an approximately 500-fold greater potency in terms of transcriptional activation of the human AHR relative to the mouse AHR in cell lines. Structure-function studies reveal that the sulfate group is an important determinant for efficient AHR activation. This is the first phase II enzymatic product identified that can significantly activate the AHR, and ligand competition binding assays indicate that I3S is a direct AHR ligand. I3S failed to activate either CAR or PXR. The physiological importance of I3S lies in the fact that it is a key uremic toxin that accumulates to high micromolar concentrations in kidney dialysis patients, but its mechanism of action is unknown. I3S represents the first identified relatively high potency endogenous AHR ligand that plays a key role in human disease progression. These studies provide evidence that the production of I3S can lead to AHR activation and altered drug metabolism. Our results also suggest that prolonged activation of the AHR by I3S may contribute to toxicity observed in kidney dialysis patients and thus represent a possible therapeutic target. PMID- 20000591 TI - Long bonds and short barriers: ionization and isomerization of alkyl nitriles. AB - Ab initio molecular orbital calculations demonstrate that ionizing alkyl nitriles produces a dramatic geometry change involving lengthening of a C-CH(2)CN bond. The experimental determination of the adiabatic ionization energy of these species is thus very difficult. In addition, there are generally low barriers for 1,2-H shift reactions in the molecular ions leading to RCHCHN(+*) and RCHCNH(+*) isomers, which makes generating pure ionized alkyl nitrile in a mass spectrometer a challenge. Threshold photoelectron spectroscopy and threshold photoelecton photoion coincidence spectroscopy were employed to study the ionization and dissociation of two alkyl nitriles, in particular, pentanenitrile and 2,2 dimethylpropanenitrile. Threshold ionization is shown to result not in the respective molecular ions, but rather in isomeric forms, resulting in dissociation thresholds that lie below the calculated adiabatic ionization energies of the two molecules. Appearance energies for all observed fragment ions are reported and compared to available literature values. Charge separation in the dissociation of doubly ionized 2,2-dimethylpropanenitrile is observed as fragment-ion time-of-flight peak broadening at high photon energies. PMID- 20000592 TI - Unusual quantum interference in the S1 state of DABCO and observation of intramolecular vibrational redistribution. AB - In this paper we report an experimental study of the time-resolved response of the molecule 1,4-diazabicyclo[2.2.2]octane (DABCO) to 266.3 nm electronic excitation of the S(1) state with a femtosecond laser. Rotational decoherence and vibrational oscillation within the S(1) state are observed. We performed state-of the-art ab initio calculations on the ground and low electronic states of the neutral molecule and the cation, which assist in the assignment of the observed photoelectron signals. Using our theoretical and spectroscopic data, the experimental findings are interpreted in terms of an unusual quantum interference between two different vibrational modes, with only the nu = 1 level of each mode being populated. PMID- 20000593 TI - Automated oligonucleotide solid-phase synthesis on nanosized silica particles using nano-on-micro assembled particle supports. AB - This article describes an original strategy to enable solid-phase oligodeoxyribonucleotide (ODN) synthesis on nanosized silica particles. It consists of the reversible immobilization of silica nanoparticles (NPs) on micrometric silica beads. The resulting assemblies, called nano-on-micro (NOM) systems, are well adapted to ODN synthesis in an automated instrument. First, NPs are derivatized with OH functions. For NOM assembly preparation, these functions react with the silanols of the microbeads under specific experimental conditions. Furthermore, OH groups allow ODN synthesis on the nanoparticles via phosphoramidite chemistry. The stability of the NOM assemblies during ODN solid phase synthesis is confirmed by scanning and transmission electron microscopy (SEM and TEM, respectively), together with dynamic light scattering analyses. Then, the release of ODN-functionalized nanoparticles is performed under mild conditions (1% NH(4)OH in water, 1 h, 60 degrees C). Our technique provides silica nanoparticles well functionalized with oligonucleotides, as demonstrated by hybridization experiments conducted with the cDNA target. PMID- 20000595 TI - Depositing ordered arrays of metal sulfide nanoparticles in nanostructures using supercritical fluid carbon dioxide. AB - Silver sulfide and cadmium sulfide nanoparticles of controllable sizes are synthesized using a water-in-hexane microemulsion method and stabilized by dodecanethiol. The stabilized metal sulfide nanoparticles can be deposited homogenously on flat substrates forming ordered 2-D arrays in supercritical fluid carbon dioxide (Sc-CO(2)). The use of Sc-CO(2) leaves the particles unaffected by dewetting effects caused by traditional solvents and produces uniform arrays. The Sc-CO(2) deposition technique is capable of filling nanoparticles in nanostructures of silicon wafers which is difficult to accomplish by conventional solvent evaporation methods. PMID- 20000594 TI - X-ray photoelectron spectroscopy analysis of gold surfaces after removal of thiolated DNA oligomers by ultraviolet/ozone treatment. AB - Well-ordered films of molecular DNA can be formed by the attachment of thiolated DNA oligonucleotides to a supporting gold substrate. The gold substrate represents a significant fraction of the total cost of preparing such films, and it is thus important to determine whether such substrates can be reused. Here, we investigate with X-ray photoelectron spectroscopy the suitability of UV/ozonolysis previously employed to remove alkanethiols from gold, for removing 40-mer, single- and double-stranded synthetic DNA. We find that while UV/O(3) can indeed remove thiolated DNA from gold slides, the treatment times required permit the implantation of additional organic contaminants. PMID- 20000596 TI - Transferrin receptor-targeted liposomes encapsulating anti-BCR-ABL siRNA or asODN for chronic myeloid leukemia treatment. AB - The present work aimed at the development and application of transferrin receptor (TrfR)-targeted sterically stabilized liposomes encapsulating anti-BCR-ABL siRNA or asODN. Transferrin was coupled to the surface of liposomes encapsulating siRNA or asODN through the postinsertion method. Cell association and internalization were assessed by flow cytometry and confocal microscopy, respectively. BCR-ABL mRNA and Bcr-Abl protein levels were evaluated by qRT-PCR and Western blot, respectively. Cell viability was assessed using the resazurin reduction method. The amount of coupled transferrin and the size and stability over time of the liposomes were very satisfactory and reproducible. The siRNA encapsulation yield was dependent on the concentration of the encapsulation buffer used (20 or 300 mM), as opposed to asODN encapsulation yield which was high for both concentrations tested. Cell association and internalization studies were performed in leukemia cell lines treated with liposomes coupled to Trf (Trf liposomes) or albumin (BSA-liposomes) or with nontargeted liposomes (NT liposomes) encapsulating fluorescently labeled siRNA (Cy3-siRNA). These experiments clearly indicated that BSA- and NT-liposomes have no ability to promote the delivery of the encapsulated nucleic acids and that the Trf-liposomes deliver the nucleic acids by a Trf receptor-dependent mechanism. The Trf liposomes encapsulating siRNA or asODN promote sequence-specific down-regulation of the BCR-ABL mRNA, although a certain extent of nonspecific sequence effects at the protein and cell viability level were observed. Overall, our results indicate that Trf-liposomes encapsulating gene silencing tools allow combining molecular and cellular targeting, which is a valuable approach for cancer treatment. PMID- 20000597 TI - Comment on comparative molecular simulation study of CO2/N2 and CH4/N2 separation in zeolites and metal-organic frameworks. PMID- 20000598 TI - Director configuration of liquid-crystal droplets encapsulated by polyelectrolytes. AB - Liquid-crystal 4-n-pentyl-4'-cyanobiphenyl (5CB) droplets dispersed in aqueous solution are prepared by the assembly of poly(styrenesulfonic acid) (PSSH) and poly(styrenesulfonate sodium) (PSSNa) at the 5CB/water interface. The micrometer sized PSSH-coated 5CB droplets in the space confinement formed by two parallel glass slides break up into submicrometer sized droplets under evaporation-induced flow. We find that the size reduction of the PSSH-coated droplets is accompanied by the bipolar-to-radial configuration transition of the 5CB in the droplets, while the PSSNa-coated 5CB droplets show no size-dependent configuration transition in the same size range. Our results suggest that the size-dependent director configuration of liquid-crystal droplets encapsulated by polyelectrolytes can be modulated by changing the interface conditions, which is important in designing liquid-crystal droplets for optical and biological applications. PMID- 20000599 TI - Regression model of partial pressures of PCBs, PAHs, and organochlorine pesticides in the Great Lakes' atmosphere. AB - The gas-phase concentrations of polychlorinated biphenyls (PCBs), polycyclic aromatic hydrocarbons (PAHs), and organochlorine pesticides have been measured at six sites around the Great Lakes every 12 days since the early 1990s as part of the Integrated Atmospheric Deposition Network. After converting the concentrations into partial pressures, data from all of the sites were combined and fitted using a multiple linear regression equation that included time (indicating the effect of a chemical's regulation), atmospheric temperature (indicating seasonality of use or release), the human population within a 25 km radius of the site (indicating the effect of urbanization) and wind speed and wind direction (indicating the source of the chemical). The atmospheric levels of lindane (gamma-HCH), DDTs, endosulfans, and chlordanes were largely related to seasonality, with much higher levels in the warm summer months. The levels of SigmaPCBs, SigmaPAHs, SigmaDDTs, and chlordanes were related to urbanization (this was a secondary factor for the latter two), a result that was unexpected for the two pesticides. The levels of only two compounds, alpha- and gamma-HCH, decreased rapidly as a function of time; conversely, most other compounds are declining at much slower rates. Wind speed and wind direction were statistically significant but unimportant variables for most of the compounds. PMID- 20000600 TI - Approach for estimating microbial growth and the biodegradation of hydrocarbon contaminants in subsoil based on field measurements: 1. Model development and verification. AB - An approach was developed to represent the microbial growth and corresponding biodegradation of hydrocarbons (HCs) during the natural attenuation process based on field measurements of in situ microbial biomass and residual HC concentrations in unsaturated subsurface soil. A kinetic model combining Monod and logistic kinetics represents microbial growth under the limitation of HCs as substrates and environmental factors at actual contaminated sites by the introduction of two new kinetic parameters, the effective rate and the self-limiting coefficient of microbial growth. The correspondence between microbial growth and the biodegradation of HCs in the soil is obtained by dividing the amount of HC and the corresponding degrading microbial groups into two classes: saturated HCs as inert components and aromatic HCs that form a contamination plume as dissolved components. The respiratory quinones were used as indicators of microbial biomass. The biodegradation capacity of contaminated sites was evaluated by the maximum microbial biomass obtained by field measurements, which is considered as the integrated results from measurements of HCs, degrading kinetics, and environmental factors at the site. The feasibility of the proposed approach was verified at two hypothetical contaminated sites. The results suggested that the proposed approach is feasible for application at actual HC-contaminated sites. PMID- 20000601 TI - Weakening C-O bonds: Ti(III), a new reagent for alcohol deoxygenation and carbonyl coupling olefination. AB - Investigations detailed herein, including density functional theory (DFT) calculations, demonstrate that the formation of either alkoxy- or hydroxy-Ti(III) complexes considerably decreases the energy of activation for C-O bond homolysis. As a consequence of this observation, we described two new synthetic applications of Nugent's reagent in organic chemistry. The first of these applications is an one-step methodology for deoxygenation-reduction of alcohols, including benzylic and allylic alcohols and 1,2-dihydroxy compounds. Additionally, we have also proved that Ti(III) is capable of mediating carbonyl coupling-olefination. In this sense, and despite the fact that for over 35 years it has been widely accepted that either Ti(II) or Ti(0) was the active species in the reductive process of the McMurry reaction, the mechanistic evidence presented proves the involvement of Ti(III) pinacolates in the deoxygenation step of the herein described Nugent's reagent-mediated McMurry olefination. This observation sheds some light on probably one of the mechanistically more complex transformations in organic chemistry. Finally, we have also proved that both of these processes can be performed catalytically in Cp(2)TiCl(2) by using trimethylsilyl chloride (TMSCl) as the final oxygen trap. PMID- 20000603 TI - Formation of organic nanoscale laminates and blends by molecular layer deposition. AB - Nanoscale organic films are important for many applications. We report on a system of molecular layer deposition that allows for the deposition of conformal organic films with thickness and composition control at the subnanometer length scale. Nanoscale polyurea films are grown on silica substrates in a layer-by layer fashion by dosing 1,4-phenylene diisocyanate (PDIC) and ethylenediamine (ED) in the gas phase. Ellipsometry measurements indicate that the film growth occurs at a constant growth rate, with film thicknesses consistent with molecular distances calculated using density functional theory. Characterization of the films by Fourier transform infrared spectroscopy and X-ray photoelectron spectroscopy reveals formation of stable polyurea films with nearly stoichiometric composition, and transmission electron microscopy indicates that the films uniformly coat the substrate surface. Subnanometer control over the film composition was demonstrated using 2,2'-thiobis(ethylamine) (TBEA) as an alternate diamine to vary the composition of the films. By substituting TBEA for ED, blended films, with homogeneous composition through the film, and nanolaminates, with discrete layers of differing film chemistry, were created. PMID- 20000602 TI - The stability of aryl carbanions derived from pyridine N-oxide: the role of resonance in stabilizing aryl anions. AB - The gas phase stability of carbanions centered at various positions on pyridine N oxide were investigated by computational and experimental methods. In addition, G3MP2 computations were completed on ring-deprotonated pyridine and N methylpyridinium. With these species, the effect of a nitrogen-centered positive charge on carbanion stability was assessed. Introduction of the nitrogen-oxide group into the benzene ring decreases the DeltaH(acid) by approximately 20 kcal/mol, but surprisingly, the effect is nearly independent of the position of the group (ortho, meta, or para). The results indicate that the N-oxide offers a balance of field, resonance, and local effects that cancels out any positional preferences. G3MP2 calculations indicate that a similar lack of positional selectivity is seen in nitrobenzene and benzonitrile. Overall, the data suggest that pi-effects are limited in phenyl anions, and as a result, ylide-like, rather than carbene-like, resonance structures are most important in the anions derived from ring deprotonation of arenes and heterocycles of these general types. PMID- 20000604 TI - Microcystin elimination during sediment contact. AB - Microcystins (MCYSTs) are a group of structurally similar toxic peptides produced by cyanobacteria ("blue-green algae") which occur frequently in surface waters worldwide. Reliable elimination is necessary when using these waters as drinking water sources. Bank filtration and artificial groundwater recharge utilize adsorption and degradation processes in the subsurface, commonly through sand and gravel aquifers, for the elimination of a wide range of substances during drinking water (pre-) treatment. To obtain parameters for estimating whether MCYST breakthrough is likely in field settings, we tested MCYST elimination in laboratory experiments (batch experiments, column experiments) under a range of conditions. Adsorption coefficients (k(d)-values) obtained from batch studies ranged from 0.2 mL/g for filter sand to 11.6 mL/g for fine grained aquifer materials with 2% fine grains (<63 microm) and 0.8% organic matter. First order degradation rates in column studies reached 1.87 d(-1) under aerobic conditions and showed high variations under anoxic conditions (<0.01-1.35 d(-1)). These results show that, next to sediment texture, redox conditions play an important role for MCYST elimination during sediment passage. Biodegradation was identified as the dominating process for MCYST elimination in sandy aquifer material. PMID- 20000605 TI - Determination of Leu side-chain conformations in excited protein states by NMR relaxation dispersion. AB - Fits of Carr-Purcell-Meiboom-Gill (CPMG) relaxation dispersion profiles allow extraction of the kinetics and thermodynamics of exchange reactions that interconvert highly populated, ground state and low populated, excited state conformers. Structural information is also available in the form of chemical shift differences between the interconverting protein states. Here we present a very simple method for extracting chi(2) rotamer distributions of Leu side chains in 'invisible' excited protein states based on measurement of their (13)C(delta1)/(13)C(delta2) chemical shifts using methyl CPMG dispersion experiments. The methodology is applied to study the protein folding reaction of the Fyn SH3 domain. A uniform chi(2) rotamer distribution is obtained for Leu residues of the unfolded state, with each Leu occupying the trans and gauche+ conformations in a 2:1 ratio. By contrast, leucines of an 'invisible' Fyn SH3 domain folding intermediate show a much more heterogeneous distribution of chi(2) rotamer populations. The experiment provides an important tool toward the quantitative characterization of both the structural and dynamics properties of states that cannot be studied by other biophysical tools. PMID- 20000606 TI - Challenges in fabrication of mesoporous carbon films with ordered cylindrical pores via phenolic oligomer self-assembly with triblock copolymers. AB - Mesoporous phenol formaldehyde (PF) polymer resin and carbon films are prepared by a solution self-assembly of PF oligomers with amphiphilic triblock copolymers. After thermopolymerization of the PF to cross-link the network, the films show an ordered morphology as determined by X-ray diffraction and grazing incidence small angle X-ray scattering (GISAXS). Our results show that the amphiphilic triblock copolymer template greatly influences the stability of the final porous mesostructures. The pyrolysis of the two-dimensional (2-D) hexagonal films with p6mm symmetry templated by Pluronic F127 yields a disordered porous structure following the template removal. Conversely, films templated by Pluronic P123 can exhibit well-ordered cylindrical pores after the template removal, but the solution composition range to yield ordered cylindrical mesopores is significantly reduced (nearly 70%) for thin films in comparison to bulk powders. We propose two dominant difficulties in fabricating well-ordered cylindrical mesopores in films: first, the stress from contraction during the pyrolysis can lead to a collapse of the mesostructure if the wall thickness is insufficient, and second, the surface wetting behavior in thin films leads to a small compositional range. PMID- 20000607 TI - Complete regioselective addition of grignard reagents to pyrazine N-oxides, toward an efficient enantioselective synthesis of substituted piperazines. AB - A conceptually new one-pot strategy for the synthesis of protected substituted piperazines via the addition of Grignard reagents to pyrazine N-oxides is presented. This strategy is high yielding (33-91% over three steps), step efficient, and fast. The synthesized N,N-diprotected piperazines are convenient to handle and allow for orthogonal deprotection at either nitrogen for selective transformations. In addition, this is a synthetic route to enantiomerically enriched piperazines by using a combination of phenyl magnesium chloride and (-) sparteine, which resulted in enantiomeric excesses up to 83%. PMID- 20000608 TI - Decomposition of naphthalene by dc gliding arc gas discharge. AB - Gliding arc discharge has been proved to be effective in treatment of gas and liquid contaminants. In this study, physical characteristics of dc gliding arc discharge and its application to naphthalene destruction are investigated with different external resistances and carrier gases. The decomposition rate increases with increasing of oxygen concentration and decreases with external resistance. This value can be achieved up to 92.3% at the external resistance of 50 kOmega in the oxygen discharge, while the highest destruction energy efficiency reaches 3.6 g (kW h)(-1) with the external resistance of 93 kOmega. Possible reaction pathways and degradation mechanisms in the plasma with different gases are proposed by qualitative analysis of postdestructed products. In the air and oxygen gliding arc discharges, the naphthalene degradation is mainly governed by reactions with oxygen-derived radicals. PMID- 20000609 TI - New insight into the formation of nitrogen sulfide: a quantum chemical study. AB - We studied the chemical mechanism for the formation of (2)NS in the interstellar medium was by using the CCSD/6-311++G(d,p) and CCSD(T)/6-311++G(3df,3pd) levels of theory. To the best of our knowledge, this is the first detailed study of the chemical mechanism for the formation of (2)NS. Several reactions proposed in this article are spin-forbidden. They were treated with the Landau-Zener theory and by the MRCI methodology. The following reactions paths proposed in this article are energetically favorable: (1) (1)NH + (2)SH --> cis-(2)HNSH --> TS1 --> trans (2)HNSH --> TS2 --> (2)H(2)NS --> TS3 --> (2)NS + H(2) and (2) (4)N + (1)SH --> (1)NSH --> TS13 --> (1)HNS --> (2)NS + (2)H. However, the latter reaction, (4)N + (1)SH --> (1)NSH, is spin-forbidden, and its probability of occuring (p(sh)) is zero. The chemical mechanism for the formation of (2)NS in the interstellar medium is now presented in more detail, which is of great importance. PMID- 20000610 TI - Thermochemistry and electronic structure of small boron clusters (B(n), n = 5-13) and their anions. AB - Thermochemical parameters of a set of small-sized neutral (B(n)) and anionic (B(n)(-)) boron clusters, with n = 5-13, were determined using coupled-cluster theory CCSD(T) calculations with the aug-cc-pVnZ (n = D, T, and Q) basis sets extrapolated to the complete basis set limit (CBS) plus addition corrections and/or G3B3 calculations. Enthalpies of formation, adiabatic electron affinities (EA), vertical (VDE), and adiabatic (ADE) detachment energies were evaluated. Our calculated EAs are in good agreement with recent experiments (values in eV): B(5) (CBS, 2.29; G3B3, 2.48; exptl., 2.33 +/- 0.02), B(6) (CBS, 2.59; G3B3, 3.23; exptl., 3.01 +/- 0.04), B(7) (CBS, 2.62; G3B3, 2.67; exptl., 2.55 +/- 0.05), B(8) (CBS, 3.02; G3B3, 3.11; exptl., 3.02 +/- 0.02), B(9) (G3B3, 3.03; exptl., 3.39 +/ 0.06), B(10) (G3B3, 2.85; exptl., 2.88 +/- 0.09), B(11) (G3B4, 3.48;, exptl., 3.43 +/- 0.01), B(12) (G3B3, 2.33; exptl., 2.21 +/- 0.04), and B(13) (G3B3, 3.62; exptl., 3.78 +/- 0.02). The difference between the calculated adiabatic electron affinity and the adiabatic detachment energy for B(6) is due to the fact that the geometry of the anion is not that of the ground-state neutral. The calculated adiabatic detachment energies to the (3)A(u), C(2h) and (1)A(g), D(2h) excited states of B(6), which have geometries similar to the (1)A(g), D(2h) state of B(6)(-), are 2.93 and 3.06 eV, in excellent agreement with experiment. The VDEs were also well reproduced by the calculations. Partitioning of the electron localization functions into pi and sigma components allows probing of the partial and local delocalization in global nonaromatic systems. The larger clusters appear to exhibit multiple aromaticity. The binding energies per atom vary in a parallel manner for both neutral and anionic series and approach the experimental value for the heat of atomization of B. The resonance energies and the normalized resonance energies are convenient indices to quantify the stabilization of a cluster of elements. PMID- 20000611 TI - Comment on "Degradability of an acrylate-linked, fluorotelomer polymer in soil". PMID- 20000612 TI - Response to comments on "Degradability of an acrylate-linked, fluorotelomer polymer in soil". PMID- 20000613 TI - Computational studies, virtual screening, and theoretical molecular models. PMID- 20000614 TI - Viscoelasticity and structure of polystyrene/fumed silica nanocomposites: filler network and hydrodynamic contributions. AB - We investigate the relationships between structure and linear viscoelasticity of a model polymer nanocomposite system based on a mixture of fumed silica nanoparticles and polystyrene. Alterations in the viscoelastic behavior are attributed to the structuring of primary silica aggregates. Above a critical filler volume fraction, a space-filling network builds up as the result of cluster aggregation, and the complex frequency-dependence of the moduli is simplified by splitting the viscoelasticity of the composites into the independent responses of the suspending polymer melt and the filler network. Specifically, we present a refinement of a two-component model recently proposed for attractive colloidal suspensions, in which hydrodynamic effects related to the presence of the filler are properly taken into account using the concept of shear stress equivalent deformation. Our approach, validated through the building of a master curve of the elastic modulus for samples of different composition, allows the estimation of the elasticity of samples in which the filler network is too tenuous to be appreciated through a simple frequency scan. In addition, the structure of the filler network is studied using both the percolation and fractal approaches, and the reliability of the critical parameters is discussed. We expect that our analysis may be useful for understanding the behavior of a wide variety of complex fluids where the elasticity of the components may be superimposed. PMID- 20000615 TI - Practical synthesis and reactivity of [3]dendralene. AB - A convenient and high-yielding three-step synthesis of the simplest branched triene, [3]dendralene, has been devised. The synthesis is robust and operationally simple, requiring no chromatography and involving no protecting groups or specialized equipment, allowing the synthesis of the volatile hydrocarbon in pure, solvent free form on a multigram scale. The stability, dimerization when stored neat, and Diels-Alder reactivity of [3]dendralene- including double cycloaddition sequences and catalytic enantioselective variant- are reported. PMID- 20000616 TI - Self-structured surface patterns on molecular azo glass films induced by laser light irradiation. AB - In this work, formation of photoinduced self-structured surface pattern and its correlation with chromophoric structures were studied by using a series of star shaped azo compounds, which exist as stable molecular glass at room temperature. For the synthesis, a star-shaped precursor (Tr-AN) was prepared by a ring-open reaction between 1, 3, 5-triglycidyl isocyanurate and N-methylaniline. The star shaped azo compounds were then synthesized through azo-coupling reactions between the precursor and diazonium salts of 4-nitroaniline, 2-methyl-4-nitroaniline, and 4-aminobenzonitrile, respectively. Through these steps, three azo compounds were obtained to bear different donor-acceptor type azo chromophores at the peripheral positions. The formation of the photoinduced self-structured patterns was investigated by irradiating solid thin films of the molecular azo glass with a uniform laser beam (532 nm, 200 mW/cm(2)) at normal incidence. For comparison, formation of surface-relief-gratings (SRGs) was also investigated by exposing the thin films to an interference pattern of the laser beams (532 nm, 80 mW/cm(2)). The results show that SRGs can be inscribed on the films of all three star-shaped azo compounds, but self-structured surface patterns is only observed on film of the azo compound containing 4-cyanoazobenzene moieties (Tr-AZ-CN) under the same irradiation condition. The electron-withdrawing groups, which control the absorption band position, show a significant influence on both the self structured pattern formation and SRG inscription rate. Under proper experimental conditions, both self-structured surface pattern and SRG can simultaneously be observed on the Tr-AZ-CN films. The observations can lead to a deeper understanding of the photoinduced effects, especially their correlation with molecular structures. PMID- 20000617 TI - Aromaticity on the fly: cyclic transition state stabilization at finite temperature. AB - We study the transition state of pericyclic reactions at elevated temperature with unbiased ab initio molecular dynamics. We find that the transition state of the intramolecular rearrangements for barbaralane and bullvalene remains aromatic at high temperature despite the significant thermal atomic motions. Structural, magnetic, and electronic properties of the dynamical transition state show the concertedness and aromatic character. Free-energy calculations also support the validity of the transition state theory for the present rearrangement reactions. The calculations demonstrate that cyclic delocalization represents a strong force to synchronize the thermal atomic motions even at high temperatures. PMID- 20000618 TI - Diversity-oriented synthesis of quinolines via Friedlander annulation reaction under mild catalytic conditions. AB - An efficient and practical method has been manifested for the diversity-oriented synthesis of quinolines via Friedlander annulation reaction for the generation of a wide range of structurally interesting and pharmacologically significant compounds by using ceric ammonium nitrate as a catalyst (10 mol %) at ambient temperature in 45 min. A variety of functional groups are introduced at various positions of the quinoline moiety, and further the diversity of the core skeleton was expanded at R(1) and R(2) positions by the synthesis of various hybrids. Initial screening of the compounds for cytotoxicity against a series of cancer cell lines showed promising results. PMID- 20000619 TI - An efficient, selective, and reducing agent-free copper catalyst for the atom transfer radical addition of halo compounds to activated olefins. AB - Efficient and selective ATRA reactions of CCl(4), CBr(4), TsCl (Ts = tosyl), or Cl(3)CCO(2)Et with activated olefins (styrene, methyl methacrylate, n-butyl methacrylate, tert-butyl methacrylate) using the Tp(tBu)Cu(NCMe) complex as a catalyst have been achieved in the absence of any reductant and with low catalyst loadings. PMID- 20000621 TI - Building robust and reliable molecular constructs: patterning, metallic contacts, and layer-by-layer assembly. AB - We describe recent progress in our laboratories to build stable complex two- and three-dimensional molecular constructs. We have introduced a simple and robust method for constructing complex molecular devices using top-down and bottom-up techniques based on self-assembled monolayers (SAMs), lithography, and site selective reactions. It has significant advantages over other methods; it is easily scaled up, affords precise nanoscale placement, and is extensible to many different materials. Several recent developments are discussed including the UV photopatterning and electron beam lithography of SAMs adsorbed on semiconductors, the site-selective deposition of metals using electroless deposition and low temperature chemical vapor deposition, and layer-by-layer assembly using covalent coupling. Optimization and further development of these techniques requires a detailed understanding of the reaction pathways involved in the lithography of SAMs and of the interaction of SAMs with metals, organometallic compounds, ions, and other compounds. PMID- 20000620 TI - Scalable, shape-specific, top-down fabrication methods for the synthesis of engineered colloidal particles. AB - The search for a method to fabricate nonspherical colloidal particles from a variety of materials is of growing interest. As the commercialization of nanotechnology continues to expand, the ability to translate particle-fabrication methods from a laboratory to an industrial scale is of increasing significance. In this feature article, we examine several of the most readily scalable top-down methods for the fabrication of such shape-specific particles and compare their capabilities with respect to particle composition, size, shape, and complexity as well as the scalability of the method. We offer an extensive examination of particle replication in nonwetting templates (PRINT) with regard to the versatility and scalability of this technique. We also detail the specific methods used in PRINT particle fabrication, including harvesting, purification, and surface-modification techniques, with an examination of both past and current methods. PMID- 20000622 TI - A Langmuir monolayer study of the interaction of E1(145-162) hepatitis G virus peptide with phospholipid membranes. AB - E1(145-162), a peptide corresponding to the structural protein E1 of the GB virus C, has been shown earlier to bind at pH 7.4 to vesicles containing 1,2 dimyiristoyl-sn-glycero-3-phospho-rac-(1-glycerol)] (DMPG) and 1,2-dimyiristoyl sn-glycero-3-phosphocholine (DMPC) phospholipids. To deepen the understanding of the interaction of E1(145-162) with the lipid membrane, in this paper, we report a detailed study of the surface properties of peptide, miscibility properties, and behavior of mixed monomolecular films of it and three phospholipids DMPG, DMPC, and 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPG). These studies were performed using the Langmuir balance by means of surface adsorption studies, surface pressure-mean molecular area compression isotherms, and penetration kinetics. The Brewster angle microscopy (BAM) was used to study the morphological properties of pure peptide and the mixed monolayers. The results show us that the peptide showed surface activity concentration dependent when injected beneath a buffered solution (HEPES/NaCl, pH 7.4). This tendency to accumulate into the air/water interface confirms its potential capacity to interact with membranes; the higher penetration of peptide into phospholipids is attained when the monolayers are in the liquid expanded state and the lipids are charged negatively maybe due to its negative electric charge that interacts with the positive global charge of the peptide sequence. The area per molecule values obtained suggested that the main arrangement structure for E1(145-162) peptide is the alpha-helical at the air-water interface that agreed with computational prediction calculations. Miscibility studies indicated that mixtures become thermodynamically favored at low peptide molar fraction. PMID- 20000623 TI - Photovoltaic devices with a low band gap polymer and CdSe nanostructures exceeding 3% efficiency. AB - We report the fabrication and measurement of solar cells approaching a power conversion efficiency of 3.2% using a low band gap conjugated polymer poly[2,6 (4,4-bis-(2-ethylhexyl)-4H-cyclopenta[2,1-b;3,4-b']dithiophene)-alt-4,7-(2,1,3 benzothiadiazole)] and CdSe nanoparticles. These devices exhibit an external quantum efficiency (EQE) of >30% in a broad range of 350-800 nm with a maximum EQE of 55% in a range of 630-720 nm. We also present certified device efficiencies of 3.13% under AM 1.5 illumination. PMID- 20000624 TI - Structure determination of grandifotane A from Khaya grandifoliola by NMR, X-ray diffraction, and ECD calculation. AB - Grandifotane A (1), a limonoid with an unprecedented carbon skeleton, was isolated from the stem bark of Khaya grandifoliola. The structure of 1 with the absolute configuration was determined by spectroscopic data, X-ray crystallography, and ECD calculation. A biogenetic route for grandifotane A (1) involving an enzymatic Baeyer-Villiger oxidation as the key step is proposed. PMID- 20000625 TI - One-pot silver-catalyzed and PIDa-mediated sequential reactions: synthesis of polysubstituted pyrroles directly from alkynoates and amines. AB - The addition/oxidative cyclization of alkynes with amines in the presence of AgBF(4) catalyst and PIDA oxidant leads to polysubstituted pyrroles. The reaction corresponds to the construction of a pyrrole fragment, which also provides a new way to the formation of C-C bonds. PMID- 20000626 TI - Luminescent platinum complexes with terdentate ligands forming 6-membered chelate rings: advantageous and deleterious effects in N--N--N and N--C--N-coordinated complexes. AB - Platinum(II) complexes of the form [PtL(n)Cl](+) are reported, containing the N- N--N-coordinating ligands 2,6-di(8-quinolyl)pyridine (L(1)), 2,6-di(8-quinolyl)-4 methoxypyridine (L(2)), or 2,6-di(7-aza-indolyl)-pyridine (L(3)). Metathesis of the chloride co-ligand in [PtL(1)Cl](+) can be accomplished under mild conditions, as exemplified by the formation of the complexes [PtL(1)OMe](+) and [PtL(1)(C[triple bond]C-tfp)](+), in which L(1) remains bound as a terdentate ligand {HC[triple bond]C-tfp = 3,5-bis(trifluoromethyl)-phenylacetylene}. An N--C -N-coordinated, cyclometalated analogue of [PtL(1)Cl](+) has also been prepared, namely, PtL(4)Cl where HL(4) is 1,3-di(8-quinolyl)benzene. The common feature among the six new complexes described here is that they contain 6-membered chelate rings, rather than the usual 5-membered rings that form when more common N--N--N ligands, such as 2,2':6',2''-terpyridine (tpy), bind to Pt(II). All the quinolyl-based complexes are phosphorescent in solution at room temperature, with quantum yields up to 4%. This contrasts with the well-established lack of emission from [Pt(tpy)Cl](+) under these conditions. Density functional theory calculations suggest that the improvement may stem, at least in part, from the relief of ring strain associated with the larger chelate ring size, leading to a more optimal bite angle at the metal, close to 180 degrees , and hence to a stronger ligand field. Consideration of the luminescence parameters, including data at 77 K, together with absorption and electrochemical data and the results of TD-DFT calculations, suggests that the lowest-lying singlet states have metal to-ligand charge-transfer (MLCT) character, but that the triplet state from which emission occurs has more predominant ligand-centered character. The azaindolyl complex [PtL(3)Cl](+) is not emissive at room temperature, apparently owing to a particularly small radiative rate constant. The cyclometalated complex PtL(4)Cl emits at lower energy than [PtL(1)Cl](+) but, in this case, the luminescence quantum yield is inferior to related complexes with 5-membered chelate rings. PMID- 20000627 TI - Indium chalcogenido clusters coordinated by 1,10-phenanthroline, [InQ(phen)Cl]4 (Q = Se, S), a simplified model of inorganic-organic hybrid material with cluster to-ligand charge transfer. AB - Most metal-1,10-phenanthroline-complex-sensitized inorganic semiconductors are multicomponent materials. This fundamental study was devoted to the single-phase materials fused with moieties of the phen complex and inorganic chalcogenide. Compounds 1 and 2 are new types of cubic clusters with cluster-to-ligand charge transfer. PMID- 20000628 TI - Cyanide anion binding by a triarylborane at the outer rim of a cyclometalated ruthenium(II) cationic complex. AB - As part of our ongoing interest in the design of boron-based cyanide anion receptors, we have synthesized a triaryl borane decorated by a cationic Ru(II) complex and have investigated its anion binding properties. This new borane, [(2,2'-bpy)Ru(kappa-C,N-2-(dimesitylborylphenyl)pyridinato)]OTf ([2]OTf), binds both fluoride and cyanide anions in organic solvents to afford 2-F and 2-CN whose crystal structures have been determined. UV-vis titrations in 9/1 CHCl(3)/DMF (vol.) afforded K((F(-))) = 1.1(+/-0.1) x 10(4) M(-1) and K((CN(-))) = 3.0(+/ 1.0) x 10(6) M(-1) indicating that [2](+) has a higher affinity for cyanide than for fluoride in this solvent mixture. These elevated binding constants show that the cationic Ru(II) complex increases the anion affinity of these complexes via Coulombic and inductive effects. The UV-vis spectral changes which accompany either fluoride or cyanide binding to the boron center are similar and include a 30 nm bathochromic shift of the metal-to-ligand charge transfer band. This shift is attributed to an increase in the donor ability of the boron-substituted phenylpyridine ligand upon anion binding to the boron center. Accordingly, cyclic voltammetry revealed that the Ru(II/III) redox couple of [2]OTf (E(1/2) = +0.051 V vs Fc/Fc(+)) undergoes a cathodic shift upon F(-) (DeltaE(1/2) = -0.242 V vs Fc/Fc(+)) or CN(-) (DeltaE(1/2) = -0.198 V vs Fc/Fc(+)) binding. PMID- 20000629 TI - Facile disassembly of amyloid fibrils using gemini surfactant micelles. AB - The accumulation of a peptide of 38-43 amino acids, in the form of fibrillar plaques, was one of the essential reasons for Alzheimer's disease (AD). Discovering an agent that is able to disassemble and clear disease-associated Abeta peptide fibrils from the brains of AD patients would have critical implications not only in understanding the dynamic process of peptide aggregation but also in the development of therapeutic strategies for AD. This study reported a new finding that cationic gemini surfactant C(12)C(6)C(12)Br(2) micelles can effectively disassemble the Abeta(1-40) fibrils in vitro. Systematic comparisons with other surfactants using ThT fluorescence, AFM, and FTIR techniques suggested that the disassembly effectiveness of gemini surfactant micelles arises from their special molecular structure (i.e., positively bicharged head and twin hydrophobic chains). To track the disassembly process, systematic cryoTEM characterization was also done, which suggested a three-stage disassembly process: (i) Spherical micelles are first absorbed onto the Abeta fibrils because of attractive electrostatic interaction. (ii) Elongated fibrils then disintegrate into short pieces and form nanoscopic aggregates via synergistic hydrophobic and electrostatic interactions. (iii) Finally, complete disaggregation of fibrils and dynamic reassembly result in the formation of peptide/surfactant complexes. PMID- 20000630 TI - Reversible "closing" of an electrode interface functionalized with a polymer brush by an electrochemical signal. AB - The poly(4-vinyl pyridine) (P4VP)-brush-modified indium tin oxide (ITO) electrode was used to switch reversibly the interfacial activity by the electrochemical signal. The application of an external potential (-0.85 V vs Ag|AgCl|KCl, 3M) that electrochemically reduced O(2) resulted in the concomitant consumption of hydrogen ions at the electrode interface, thus yielding a higher pH value and triggering the restructuring of the P4VP brush on the electrode surface. The initial swollen state of the protonated P4VP brush (pH 4.4) was permeable to the anionic [Fe(CN)(6)](4-) redox species, but the electrochemically produced local pH of 9.1 resulted in the deprotonation of the polymer brush. The produced hydrophobic shrunken state of the polymer brush was impermeable to the anionic redox species, thus fully inhibiting its redox process at the electrode surface. The interface's return to the electrochemically active state was achieved by disconnecting the applied potential, followed by stirring the electrolyte solution or by slow diffusional exchange of the electrode-adjacent thin layer with the bulk solution. The developed approach allowed the electrochemically triggered inhibition ("closing") of the electrode interface. The application of this approach to different interfacial systems will allow the use of various switchable electrodes that are useful for biosensors and biofuel cells with externally controlled activity. Further use of this concept was suggested for electrochemically controlled chemical actuators (e.g. operating as electroswitchable drug releasers). PMID- 20000631 TI - Colloidosome-based synthesis of a multifunctional nanostructure of silver and hollow iron oxide nanoparticles. AB - Nanoparticles that self-assemble on a liquid-liquid interface serve as the building block for making heterodimeric nanostructures. Specifically, hollow iron oxide nanoparticles within hexane form colloidosomes in the aqueous solution of silver nitrate, and iron oxide exposed to the aqueous phase catalyzes the reduction of silver ions to afford a heterodimer of silver and hollow iron oxide nanoparticles. Transmission electron microscopy, selected area electron diffraction, energy-dispersive X-ray spectrometry, X-ray diffraction, UV-vis spectroscopy, and SQUID were used to characterize the heterodimers. Interestingly, the formation of silver nanoparticles helps the removal of spinglass layer on the hollow iron oxide nanoparticles. This work demonstrates a powerful yet convenient strategy for producing sophisticated, multifunctional nanostructures. PMID- 20000632 TI - Mechanistic studies of human spermine oxidase: kinetic mechanism and pH effects. AB - In mammalian cells, the flavoprotein spermine oxidase (SMO) catalyzes the oxidation of spermine to spermidine and 3-aminopropanal. Mechanistic studies have been conducted with the recombinant human enzyme. The initial velocity pattern in which the ratio between the concentrations of spermine and oxygen is kept constant establishes the steady-state kinetic pattern as ping-pong. Reduction of SMO by spermine in the absence of oxygen is biphasic. The rate constant for the rapid phase varies with the substrate concentration, with a limiting value (k(3)) of 49 s(-1) and an apparent K(d) value of 48 microM at pH 8.3. The rate constant for the slow step is independent of the spermine concentration, with a value of 5.5 s(-1), comparable to the k(cat) value of 6.6 s(-1). The kinetics of the oxidative half-reaction depend on the aging time after the spermine and enzyme are mixed in a double-mixing experiment. At an aging time of 6 s, the reaction is monophasic with a second-order rate constant of 4.2 mM(-1) s(-1). At an aging time of 0.3 s, the reaction is biphasic with two second-order constants equal to 4.0 and 40 mM(-1) s(-1). Neither is equal to the k(cat)/K(O(2)) value of 13 mM( 1) s(-1). These results establish the existence of more than one pathway for the reaction of the reduced flavin intermediate with oxygen. The k(cat)/K(M) value for spermine exhibits a bell-shaped pH profile, with an average pK(a) value of 8.3. This profile is consistent with the active form of spermine having three charged nitrogens. The pH profile for k(3) shows a pK(a) value of 7.4 for a group that must be unprotonated. The pK(i)-pH profiles for the competitive inhibitors N,N'-dibenzylbutane-1,4-diamine and spermidine show that the fully protonated forms of the inhibitors and the unprotonated form of an amino acid residue with a pK(a) of approximately 7.4 in the active site are preferred for binding. PMID- 20000633 TI - Surface area characterization of obliquely deposited metal oxide nanostructured thin films. AB - The glancing angle deposition (GLAD) technique is used to fabricate nanostructured thin films with high surface area. Quantifying this property is important for optimizing GLAD-based device performance. Our group has used high sensitivity krypton gas adsorption and the complementary technique of cyclic voltammetry to measure surface area as a function of deposition angle, thickness, and morphological characteristics for several metal oxide thin films. In this work, we studied amorphous titanium dioxide (TiO(2)), amorphous silicon dioxide (SiO(2)), and polycrystalline indium tin oxide (ITO) nanostructures with vertical and helical post morphologies over a range of oblique deposition angles from 0 to 86 degrees. Krypton gas sorption isotherms, evaluated using the Brunauer-Emmettt Teller (BET) method, revealed maximum surface area enhancements of 880 +/- 110, 980 +/- 125, and 210 +/- 30 times the footprint area (equivalently 300 +/- 40, 570 +/- 70, and 50 +/- 6 m(2) g(-1)) for vertical posts TiO(2), SiO(2), and ITO. We also applied the cyclic voltammetry technique to these ITO films and observed the same overall trends as seen with the BET method. In addition, we applied the BET method to the measurement of helical films and found that the surface area trend was shifted with respect to that of vertical post films. This revealed the important influence of the substrate rotation rate and film morphology on surface properties. Finally, we showed that the surface area scales linearly with film thickness, with slopes of 730 +/- 35 to 235 +/- 10 m(2) m(-2) microm(-1) found for titania vertical post films deposited at angles from 70 to 85 degrees. This characterization effort will allow for the optimization of solar, photonic, and sensing devices fabricated from thin metal oxide films using GLAD. PMID- 20000634 TI - Controllable synthetic molecular channels: biomimetic ammonia switch. AB - We use molecular dynamics simulations combined with iterative screening to test if one can design mechanically controllable and selective molecular pores. The first model pore is formed by two stacked carbon nanocones connected by aliphatic chains at their open tips, in analogy to aquaporins. It turns out that when one nanocone is gradually rotated with respect to the other, the molecular chains alter the size of the nanopore formed at the cone tips and control the flow rates of liquid pentane through it. The second model pore is formed by two carbon nanotubes joined by a cylindrical structure of antiparallel peptides. By application of a torque to one of the nanotubes, while holding the other, we can reversibly fold the peptides into forward or backward-twisted barrels. We have modified the internal residues in these barrels to make these pores selective and controllable. Eventually, we found a nanopore that in the two folded configurations has very different transmission rates for hydrated NH(3) molecules. PMID- 20000635 TI - Stability of the DMF-protected Au nanoclusters: photochemical, dispersion, and thermal properties. AB - We have reported the synthesis of dimethylformamide (DMF)-protected gold nanoclusters using a surfactant-free DMF reduction method. DMF-protected gold nanoclusters (Au NCs) are obtained without the formation of gold nanoparticles and bulk metals as byproducts using a hot injection process for the homogeneous reduction. The as-prepared DMF-protected Au NCs were a mixture of various-sized Au NCs with a cluster number of less than 20 including at least Au(8) and Au(13). The photoluminescence emission from Au(8) and Au(13) was confirmed in the photoluminescence spectra. The Au NCs are stabilized with DMF molecules through the interaction of amide groups of DMF with Au NCs. DMF-protected Au NCs in solution were found to have high thermal stability, high dispersion stability in various solvents, and high photochemical stability. The DMF-protected Au NCs dispersed well for at least a month in various solvents such as water, acid (pH 2), alkali (pH 12) and 0.5 M NaCl aqueous solution, and methanol without further surface modification. The thermal stability of DMF-protected Au NCs was approximately 150 degrees C, which was comparable to that of thiolate-protected Au NCs. The photobleaching of Au NCs in water gradually occurred under UV light irradiation (356 nm, 1.3 mW/cm(2)) because of the photoinduced oxidation of Au NCs. After 8 h irradiation, the fluorescence intensity slowly decreased to approximately 50% of the maximum and to approximately 20% after 96 h under the present condition, compared to the photobleaching of CdSe semiconductor quantum dots. We also found that the fluorescence intensity remained to be about 30% of the maximum even in the presence of concentrated 30% H(2)O(2). These findings demonstrate that the photobleaching process under the UV irradiation is effectively suppressed for DMF-protected Au NCs. PMID- 20000636 TI - Fabrication of biomimetic superhydrophobic surface on engineering materials by a simple electroless galvanic deposition method. AB - We have reported an easy means in this paper to imitate the "lotus leaf" by constructing a superhydrophobic surface through a process combining both electroless galvanic deposition and self-assembly of n-octadecanethiol. Superhydrophobicity with a static water contact angle of about 169 +/- 2 degrees and a sliding angle of 0 +/- 2 degrees was achieved. Both the surface chemical compositions and morphological structures were analyzed. We have obtained a feather-like surface structure, and the thickness of the Ag film is about 10-30 microm. The stability of the superhydrophobic surface was tested under the following three conditions: (1) pH value from 1 to 13; (2) after freezing treatment at -20 degrees C; (3) at ambient temperature. It shows a notable stability in that the contact angle of the sample still remained higher than 150 degrees in different conditions. It can be concluded that our approach can provide an alternative way to fabricate stable superhydrophobic materials. PMID- 20000637 TI - A hybrid, de novo based, genome-wide database search approach applied to the sea urchin neuropeptidome. AB - Peptidomics is the identification and study of the in vivo biologically active peptide profile. A combination of high performance liquid chromatography, mass spectrometry, and bioinformatics tools such as database search engines are commonly used to perform the analysis. We report a methodology based on a database system holding the completed translated genome, whereby de novo sequencing and genome-wide database searching are combined. The methodology was applied to the sea urchin neuropeptidome resulting in a 30% increase in identification rate. PMID- 20000638 TI - Trapping and stabilization of integral membrane proteins by hydrophobically grafted glucose-based telomers. AB - Amphipols (APols) are short amphipathic polymers designed to adsorb onto the transmembrane surface of membrane proteins, keeping them water-soluble in the absence of detergent. Current APols carry charged groups, which is a limitation for certain types of applications. This has prompted the development of totally nonionic amphiphols (NAPols). In a previous work, glucose-based NAPols synthesized by free-radical cotelomerization of hydrophilic and amphiphilic monomers proved to be able to keep membrane proteins soluble (Sharma et al. Langmuir 2008, 24, 13581-13590). This provided a proof of principle, but the cumbersome synthesis prevented large-scale production and any detailed biochemical studies. In the present work, we describe a new synthesis route for NAPols based on grafting alkyl chains onto a glucosylated homotelomer. The NAPols thus prepared are highly water soluble. In aqueous solutions, they assemble into small, homogeneous particles similar to those formed by ionic APols. Two model membrane proteins, bacteriorhodopsin and the transmembrane domain of OmpA, form with NAPols small, well-defined water-soluble complexes whose size is comparable to that observed with ionic APols. Complexation by NAPols strongly stabilizes bacteriorhodopsin against denaturation. Glucosylated NAPols thus appear as a promising alternative to ionic APols for such applications as ion-exchange chromatography, isoelectrofocusing, and, possibly, structural approaches such as NMR and crystallography. PMID- 20000639 TI - Multichannel simultaneous measurements of single-molecule translocation in alpha hemolysin nanopore array. AB - We present a microarray system that enables simultaneous monitoring of multiple ionic currents through transmembrane alpha-hemolysin nanopores arrayed at bilayer lipid membranes. We applied the self-assembling ability of lipid molecules interfaced between an aqueous solution and organic solvent to induce bilayer membrane formation at a microfluidic device; the device consists of a hydrophobic polymer film that serves to suspend the lipid-containing solvent at micrometer sized apertures as well as to separate the aqueous solution into two chambers. In this study, we confirmed that expeditious and reproducible bilayer formation is realized by control of the composition of the solvent, a mixture of n-decane and 1-hexanol, which permits simultaneous incorporation of the alpha-hemolysin nanopores to the membrane array. Monitoring the eight wells on the array at once, we obtained a maximum of four relevant, synchronous signals of translocating ionic current through the nanopores. The system was also able to detect translocation events of nucleic acid molecules through the pore via the profile of a blocked current, promising its potential for high-throughput applications. PMID- 20000640 TI - Sensitive bifunctional aptamer-based electrochemical biosensor for small molecules and protein. AB - In this paper, a bifunctional electrochemical biosensor for highly sensitive detection of small molecule (adenosine) or protein (lysozyme) was developed. Two aptamer units for adenosine and lysozyme were immobilized on the gold electrode by the formation of DNA/DNA duplex. The detection of adenosine or lysozyme could be carried out by virtue of switching structures of aptamers from DNA/DNA duplex to DNA/target complex. The change of the interfacial feature of the electrode was characterized by cyclic voltammertic (CV) response of surface-bound [Ru(NH(3))(6)](3+). On the other hand, DNA functionalized Au nanoparticles (DNA AuNPs) were used to enhance the sensitivity of the aptasensor because DNA-AuNPs modified interface could load more [Ru(NH(3))(6)](3+) cations. Thus, the assembly of two aptamer-contained DNA strands integrated with the DNA-AuNPs amplification not only improves the sensitivity of the electrochemical aptasensor but also presents a simple and general model for bifunctional aptasensor. The proposed aptasensor has low detection limit (0.02 nM for adenosine and 0.01 microg mL(-1) for lysozyme) and exhibits several advantages such as high sensitivity and bifunctional recognition. PMID- 20000641 TI - Quantification of extraneous carbon during compound specific radiocarbon analysis of black carbon. AB - Radiocarbon ((14)C) is a radioactive isotope that is useful for determining the age and cycling of carbon-based materials in the Earth system. Compound specific radiocarbon analysis (CSRA) provides powerful insight into the turnover of individual components that make up the carbon cycle. Extraneous or nonspecific background carbon (C(ex)) is added during sample processing and subsequent isolation of CSRA samples. Here, we evaluate the quantity and radiocarbon signature of C(ex) added from two sources: preparative capillary gas chromatography (PCGC, C(PCGC)) and chemical preparation of CSRA of black carbon samples (C(chemistry)). We evaluated the blank directly using process blanks and indirectly by quantifying the difference in the isotopic composition between processed and unprocessed samples for a range of sample sizes. The direct and indirect assessment of C(chemistry+PCGC) agree, both in magnitude and radiocarbon value (1.1 +/- 0.5 microg of C, fraction modern = 0.2). Half of the C(ex) is introduced before PCGC isolation, likely from coeluting compounds in solvents used in the extraction method. The magnitude of propagated uncertainties of CSRA samples are a function of sample size and collection duration. Small samples collected for a brief amount of time have a smaller propagated (14)C uncertainty than larger samples collected for a longer period of time. CSRA users are cautioned to consider the magnitude of uncertainty they require for their system of interest, to frequently evaluate the magnitude of C(ex) added during sampling processing, and to avoid isolating samples < or = 5 microg of carbon. PMID- 20000642 TI - Development and single-laboratory validation of a pseudofunctional biosensor immunoassay for the detection of the okadaic acid group of toxins. AB - A rapid analytical optical biosensor-based immunoassay was developed and validated for the detection of okadaic acid (OA) and its structurally related toxins from shellfish matrix. The assay utilizes a monoclonal antibody which binds to the OA group of toxins in order of their toxicities, resulting in a pseudofunctional assay. Single-laboratory validation of the assay for quantitative detection of OA determined that it has an action limit of 120 microg/kg, a limit of detection of 31 microg/kg, and a working range of 31-174 microg/kg. The midpoint on the standard matrix calibration curve is 80 microg/kg, half the current regulatory limit. Inter- and intra-assay studies of negative mussel samples spiked with various OA concentrations produced average coefficient of variation (CV) and standard deviation (SD) values of 7.9 and 10.1, respectively. The assay was also validated to confirm the ability to accurately codetect and quantify dinophysistoxin-1 (DTX-1), DTX-2, and DTX-3 from shellfish matrix. Alkaline hydrolysis was not required for the detection of DTX-3 from matrix. Excellent correlations with the data generated by the biosensor method and liquid chromatography/tandem mass spectrometry (LC/MS/MS) were obtained using a certified reference material (R(2) = 0.99), laboratory reference material, and naturally contaminated mussel samples (R(2) = 0.97). This new procedure could be used as a rapid screening procedure replacing animal-based tests for DSP toxins. PMID- 20000643 TI - Development of new HPLC chiral stationary phases based on native and derivatized cyclofructans. AB - An unusual class of chiral selectors, cyclofructans, is introduced for the first time as bonded chiral stationary phases. Compared to native cyclofructans (CFs), which have rather limited capabilities as chiral selectors, aliphatic- and aromatic-functionalized CF6s possess unique and very different enantiomeric selectivities. Indeed, they are shown to separate a very broad range of racemic compounds. In particular, aliphatic-derivatized CF6s with a low substitution degree baseline separate all tested chiral primary amines. It appears that partial derivatization on the CF6 molecule disrupts the molecular internal hydrogen bonding, thereby making the core of the molecule more accessible. In contrast, highly aromatic-functionalized CF6 stationary phases lose most of the enantioselective capabilities toward primary amines, however they gain broad selectivity for most other types of analytes. This class of stationary phases also demonstrates high "loadability" and therefore has great potential for preparative separations. The variations in enantiomeric selectivity often can be correlated with distinct structural features of the selector. The separations occur predominantly in the presence of organic solvents. PMID- 20000644 TI - Visible chromophore phosphines as functional elements of luminescent metallopolymers. AB - A general modular route to fluorophore-linked diphosphines from (4 iodophenyl)P(O)Ph(2) is described. The preparation of chromophoric diphosphine (LHP1) proceeded readily using this route. LHP1 was employed to prepare luminescent metallopolymers of platinum (P1) and palladium (P2) exhibiting modest degrees of polymerization (13-14) and extents of polymerization ( approximately 97%). P1 and P2 appear to be the first metallopolymers polymerized via a visible absorbing/emitting diphosphine. The photophysical properties of these novel materials are discussed. PMID- 20000645 TI - Syntheses, crystal and electronic structures, and physical properties of two quaternary chalcogenides: La(4)FeSb(2)Q(10) (Q = S, Se). AB - Two new quaternary chalcogenides, La(4)FeSb(2)S(10) and La(4)FeSb(2)Se(10), have been synthesized from the stoichiometric mixture of elements by solid-state reactions at 1100 degrees C. The compounds crystallize in the orthorhombic space group Pbcm with a = 15.066(4) A, b = 7.590(2) A, c = 13.341(4) A, and Z = 4 and a = 15.596(5) A, b = 7.869(2) A, c = 13.960(4) A, and Z = 4, respectively. These structures represent an unique three-dimensional network, in which SbQ(3) trigonal pyramids (Sb-S < 2.60 A, Sb-Se < 2.80 A) are connected via a relatively weak Sb-Q bond (Sb-S approximately 2.90 A, Sb-Se approximately 3.00 A) in a novel teeter-totter (SbQ(4))(n) chain motif. The theoretical studies confirm the Sb-Q bonding interactions within such teeter-totter chains. Their optical band gaps are measured to be 1.00 and 0.85 eV. At room temperature, their electrical conductivities are about 10(-4) S/cm. Both compounds display antiferromagnetic interactions between Fe centers, and their effective magnetic moments are 5.25 and 6.17 micro(B), respectively. PMID- 20000646 TI - Luminescence properties and energy transfer of site-sensitive Ca(6-x-y)Mg(x z)(PO(4))(4):Eu(y)(2+),Mn(z)(2+) phosphors and their application to near-UV LED based white LEDs. AB - On the basis of the structural information that the host material has excellent charge stabilization, blue-emitting Ca(6-x-y)Mg(x)(PO(4))(4):Eu(y)(2+) (CMP:Eu(2+)) phosphors were synthesized and systematically optimized, and their photoluminescence (PL) properties were evaluated. Depending upon the amount of Mg added, the emission efficiency of the phosphors could be enhanced. The substitution of Eu(2+) affected their maximum wavelength (lambda(max)) and thermal stability because the substitution site of Eu(2+) could be varied. To obtain single-phase two-color-emitting phosphors, we incorporated Mn(2+) into CMP:Eu(2+) phosphors. Weak red emission resulting from the forbidden transition of Mn(2+) could be enhanced by the energy transfer from Eu(2+) to Mn(2+) that occurs because of the spectral overlap between the photoluminescence excitation (PLE) spectrum of Mn(2+) and the PL spectrum of Eu(2+). The energy transfer process was confirmed by the luminescence spectra, energy transfer efficiency, and decay curve of the phosphors. Finally, the optimized Ca(6-x-y)Mg(x z)(PO(4))(4):Eu(y)(2+),Mn(z)(2+) (CMP:Eu(2+),Mn(2+)) phosphors were applied with green emitting Ca(2)MgSi(2)O(7):Eu(2+) (CMS:Eu(2+)) phosphors to ultraviolet (UV) light emitting diode (LED)-pumped white LEDs. The CMS:Eu(2+)-mixed CMP:Eu(2+), Mn(2+)-based white LEDs showed an excellent color rendering index (CRI) of 98 because of the broader emission band and more stable color coordinates than those of commercial Y(3)Al(5)O(12):Ce(3+) (YAG:Ce(3+))-based white LEDs under a forward bias current of 20 mA. The fabricated white LEDs showed very bright natural white light that had the color coordinate of (0.3288, 0.3401), and thus CMP:Eu(2+),Mn(2+) could be regarded as a good candidate for UV LED-based white LEDs. PMID- 20000647 TI - Preparation, facile deprotonation, and rapid H/D exchange of the mu-hydride diiron model complexes of the [FeFe]-hydrogenase containing a pendant amine in a chelating diphosphine ligand. AB - The CO-displacement of [(mu-pdt)Fe(2)(CO)(6)] with (Ph(2)PCH(2))(2)N(n-Pr) in refluxing toluene gave an unsymmetrical chelating complex [(mu pdt){Fe(CO)(3)}{Fe(CO)(kappa(2)-Ph(2)PCH(2)N(n-Pr)CH(2)PPh(2)}] (1) as a major product, together with a small amount of the symmetrical intramolecular bridging complex [(mu-pdt){mu-Ph(2)PCH(2)N(n-Pr)CH(2)PPh(2)}{Fe(CO)(2)}(2)] (2) and the intermolecular bridging complex [{mu,kappa(1),kappa(1)-Ph(2)PCH(2)N(n Pr)CH(2)PPh(2)}{(mu-pdt)Fe(2)(CO)(5)}(2)] (3). In contrast, the reaction of [(mu pdt)Fe(2)(CO)(6)] with (Ph(2)PCH(2))(2)NR (R = n-Pr, Ph) afforded the intermolecular bridging isomers 3 and 4 in the presence of a CO-removing reagent Me(3)NO.2H(2)O in CH(3)CN at room temperature. The molecular structures of 1, 3, and 4, as well as the doubly protonated complex [1(H(N)H(mu))](OTf)(2)] were determined by X-ray analyses. The protonation processes of 1 with HBF(4).Et(2)O and HOTf were studied in different solvents. The presence of the H(mu)...H(N) interaction in [1(H(N)H(mu))](2+) was studied by relaxation time T(1) and spin saturation transfer measurements. The mu-hydride of [1(H(mu))](+) and [1(H(N)H(mu))](2+) undergo facile deprotonation with aniline and rapid H/D exchange with deuterons in solution. In contrast, neither deprotonation nor H/D exchange was detected for [(mu-H)(mu-pdt){Fe(CO)(3)}{Fe(CO)(kappa(2)-dppp)}](+) ([5(H(mu))](+), dppp = Ph(2)PCH(2)CH(2)CH(2)PPh(2)) without internal base. PMID- 20000648 TI - Synthesis, characterization, and magnetism of divalent aryl transition-metal complexes of the simplest dialkylamide, NMe(2): rare T-shaped coordination at chromium. AB - The synthesis and characterization of a series of first-row aryl transition metal derivatives of the simplest dialkylamido ligand NMe(2) are reported. The complexes Cr{Ar'Cr(mu-NMe(2))(2)}(2) (1) and {Ar'M(mu-NMe(2))}(2) (M = Mn (2), Fe (3); Ar' = C(6)H(3)-2,6-(C(6)H(3)-2,6-(i)Pr(2))(2)) were obtained by reaction of the aryl metal halides {Ar'M(mu-X)}(2) (M = Cr, X = Cl; M = Fe, X = Br) or {Li(THF)Ar'MnI(2)}(2) with LiNMe(2) in a 1:2 ratio. A similar reaction of {Ar(#)Co(mu-I)}(2) (Ar(#) = C(6)H(3)-2,6-(C(6)H(2)-2,4,6-Me(3))(2)) and LiNMe(2) in hexanes gave the unusual complex {Ar(#)Co(mu-I)(eta(1)-CH(2) horizontal lineNCH(3))}(2) (4), in which the NMe(2) ligand is dehydrogenated to afford a complexed imine. Complexes 1-4 were characterized by X-ray crystallography, UV vis spectroscopy, and magnetic measurements. In the unique trinuclear complex 1, the central chromium(II) ion is bound to four NMe(2) groups in a square planar fashion. The NMe(2) groups also bridge to the two outer chromium(II) ions, which are bound to a terminal Ar' group to yield a rare example of three-coordinate T shaped geometry at these atoms. In the dimers 2 and 3, each metal center is coordinated to a terminal terphenyl ligand and two bridging NMe(2) groups to give a distorted trigonal planar geometry. In contrast, the reaction of LiNMe(2) with {Ar(#)Co(mu-I)}(2) in a 2:1 ratio did not yield an amido product; instead, the NMe(2) ligand underwent hydrogen elimination. As a result, in the dimeric structure of 4, each cobalt ion is coordinated to a terphenyl ligand, two bridging iodides, and a neutral methylimine ligand, CH(2) horizontal lineNCH(3), to yield a very distorted tetrahedral cobalt(II) coordination environment. The magnetic properties of 1-4 revealed antiferromagnetic exchange coupling between the metal ions with J = -47(1) cm(-1) and J(13) = -25(1) cm(-1) for 1, J = -38(1) cm(-1) for 2, J = -75(3) cm(-1) for 3, and J = -32(4) cm(-1) for 4; the latter compound exhibited an unusually large temperature independent contribution to its molar magnetic susceptibility. PMID- 20000649 TI - Thiacalix[4]arene-supported planar Ln(4) (Ln = Tb(III), Dy(III)) clusters: toward luminescent and magnetic bifunctional materials. AB - This paper reports the syntheses, crystal structures, and luminescent and magnetic properties of four tetranuclear Tb(III) (1 and 3) and Dy(III) (2 and 4) complexes supported by p-phenylthiacalix[4]arene (H(4)PTC4A) and p-tert butylthiacalix[4]arene (H(4)TC4A). All four frameworks can be formulated as [Ln(III)(4)(PTC4A/TC4A)(2)(mu(4)-OH)Cl(3)(CH(3)OH)(2)(H(2)O)(3)], and some methanol and water solvent molecules are occupied in the interstices. The compounds are featured with a sandwichlike unit constructed by two tail-to-tail calixarene molecules and a planar tetragonal (mu(4)-OH)Ln(4) cluster. The photoluminescent analyses suggest that there is an efficient ligand-to-Ln(III) energy transfer for compounds 1-3 and H(4)PTC4A is a more efficient "antenna" than H(4)TC4A. The Dy(III) compounds exhibit slow magnetic relaxation behavior of single-molecule magnet nature. The substitution of the t-Bu group with a phenyl group at the up-rim of thiacalix[4]arene leads to different extended structures and physical properties of as-synthesized compounds. PMID- 20000650 TI - Photochemical hydroxylation of 1-methyl-9,10-anthraquinones: synthesis of 9' hydroxyaloesaponarin II. AB - Photolysis of 1-methyl-9,10-anthraquinones in the presence of oxygen yields endoperoxides that can be reduced to produce 1-hydroxymethyl-9,10-anthraquinones. The reaction proceeds in a fashion similar to that of other o-alkylphenones which yield either a 1,4-diradical or a "photoenol" upon irradiation. Anthraquinones undergo photochemistry at a wavelength where the endoperoxide is transparent, allowing its isolation. A singlet oxygen quencher had no effect on the rate of formation of the endoperoxide. The photochemical hydroxylation has been used in a total synthesis of a naturally occurring polyketide, 9'-hydroxyaloesaponarin II. PMID- 20000651 TI - Highly shape-selective synthesis, silica coating, self-assembly, and magnetic hydrogen sensing of hematite nanoparticles. AB - The open forced hydrolysis method and controllable silica growth based on bound water to polyvinylpyrrolidone molecules have been developed for the highly shape (including rhombohedra, semispheres, and rods) selective synthesis, self assembly, and uniform silica coating (in the unprecedented range of 5-200 nm) of hematite nanoparticles. The open system realizes the direct short-range self assembly of hematite semispheres in their growth process. The bound water method has been extended to coat gold nanoparticles with tunable silica shell and directly assemble the cores into one-dimensional, dimer, and trimer nanostructures during the coating process. The silica coating increases the particle stability and monodispersity even as hematite is modified into ferromagnetic Fe(3)O(4). The hematite@silica core-shell spheres are assembled into long-range ordered structures with considerable photonic bandgap for the first time due to their high monodispersity. By exploiting the hematite antiferromagnetism caused by the superexchange interaction via intervening oxygen ions that are sensitive to hydrogen, a novel hydrogen sensing based on magnetization variations is achieved in the hematite assemblies. Weakening the antiferromagnetism by reducing the hematite size and/or covering the hematite surface by silica coating suppresses the sensitivity to hydrogen, showing that the antiferromagnetic spin variations on the hematite surface are responsible for the gas sensing. PMID- 20000652 TI - Origin of stereocontrol in the construction of the 12 oxatricyclo[6.3.1.0(2,7)]dodecane ring system by Prins-pinacol reactions. AB - Polycyclic products containing the 12-oxatricyclo[6.3.1.0(2,7)]dodecane moiety having either the trans (8a-e) or cis (9a-e) relative configuration of the oxacyclic bridge and the cis angular substituents are formed stereospecifically by Prins-pinacol cyclizations of unsaturated alpha-dithianyl acetals 14a-e or 15a e. These results show that the topography (boat or chair) of the Prins cyclization of the sulfur-stabilized oxocarbenium ions generated from acetals 14a e or 15a-e is controlled by the stereoelectronic influence of the allylic substituents, with steric effects playing a minor role. A complex molecular rearrangement that is terminated by a thio-Prins-pinacol reaction is also identified. PMID- 20000653 TI - Mean excitation energies and energy deposition characteristics of bio-organic molecules. AB - We suggest a method for determination of the mean excitation energies of several biomolecules, such as amino acids, using a Bragg-like sum rule developed for molecular fragments or functional groups. Because the fragment composition of many bio-organic molecules is very similar, we find that many of them, including the amino acids, have similar mean excitation energies (approximately 70 eV, which is interestingly close to the mean excitation energy of the major component of a cell: namely, water). Differences in amino acid mean excitation energies arise from variation of the side chains (-R); addition of -CH(2)- groups decreases the mean excitation energy. PMID- 20000654 TI - Synthesis and reactions of phosphaporphyrins: reconstruction of pi-skeleton triggered by oxygenation of a core phosphorus atom. AB - The synthesis, structures, optical and redox properties, and reactivity of phosphaporphyrins are reported. The 21-phosphaporphyrin (P,N(3)-porphyrin) and 23 phospha-21-thiaporphyrin (P,S,N(2)-porphyrin) were prepared via acid-promoted dehydrative condensation between a phosphatripyrrane and the corresponding 2,5 bis[hydroxy(phenyl)methyl]heteroles followed by 2,3-dichloro-5,6 dicyanobenzoquinone oxidation. Experimental (NMR, UV-vis, and X-ray analyses) and theoretical (DFT calculations) results suggest that the 18pi aromaticity inherent in regular N(4)-porphyrins was maintained in these phosphaporphyrins. X-ray crystallography revealed a slightly distorted 18pi aromatic ring for the P,N(3) porphyrin with the phosphole and three pyrrole rings tilted from the 24-atoms mean plane by 9.6 degrees and 3.8-15.4 degrees, respectively. DFT calculations on model compounds showed that the P,X,N(2)-porphyrins (X = N, S) possess considerably small HOMO-LUMO gaps as compared with N(4)- and S,N(3)-porphyrins, which is reflected in the red-shifted absorptions, low oxidation potentials, and high reduction potentials of the phosphaporphyrins. The P-oxygenation of the P,X,N(2)-porphyrins with H(2)O(2) has been found to lead to the formation of different types of products. The 18pi P,N(3)-porphyrin was transformed into the 22pi aromatic P(O),N(3)-porphyrin accompanied by the pi extension at the peripheral C(3) bridge, whereas the 18pi P,S,N(2)-porphyrin was converted to the isophlorin-type 20pi antiaromatic P(O),S,N(2)-porphyrin. In both of the reactions, simple P-oxygenated 18pi P(O),X,N(2)-porphyrins were formed as the initial products, which were subsequently transformed into the 22pi or 20pi porphyrins. The two reaction courses from 18pi to 20pi/22pi are apparently determined by the combination of the core heteroatoms (i.e., P,N(3) or P,S,N(2)) and the structure of the peripherally fused carbocycles. The present results demonstrate that the incorporation of a phosphorus atom into the core is not only a highly promising way to modify the fundamental properties of the porphyrin 18pi system but also a reliable tool to stabilize uncommon 22pi and 20pi systems through the chemical modifications at the core phosphorus atom. PMID- 20000655 TI - Neurotrophic effects of tianeptine on hippocampal neurons: a proteomic approach. AB - Tianeptine, an atypical tricyclic antidepressant with unique characteristics, can improve memory and prevent stress-induced hippocampal damage. It has neuroplastic and neurotrophic effects on hippocampal neurons and can prevent dendritic atrophy of the hippocampus in certain pathological conditions. To obtain a better understanding of the underlying mechanisms, we performed a proteomic analysis on tianeptine-treated hippocampal neurons. Primary hippocampal neurons were prepared from fetal Sprague-Dawley rats, eliminating glia cells by addition of cytosine beta-D-arabinofuranoside at day 2 in vitro (DIV2). The neurons were treated with tianeptine (10 microg/mL) or vehicle at DIV3, then harvested at DIV4 or DIV9 for immunocytochemical analysis of, respectively, neurite outgrowth or synapse formation. A proteomics analysis was performed on DIV4 neurons and the data were confirmed by Western blot analysis. Using specific markers, we demonstrated that tianeptine can augment neurite growth and promote synaptic contacts in cultured hippocampal neurons. The proteomics analysis identified 11 differentially expressed proteins, with roles in neurite growth, metabolism of neurotrophic substances, synaptogenesis, and synaptic activity homeostasis. The data shed light on the mechanisms underlying the neurotrophic effect of tianeptine observed in both animal studies and the clinic. PMID- 20000656 TI - Full state-resolved energy gain profiles of CO2 (J = 2-80) from collisions of highly vibrationally excited molecules. 1. Relaxation of pyrazine (E = 37900 cm( 1)). AB - State-resolved energy gain profiles for "strong" and "weak" collisions of pyrazine (E = 37900 cm(-1)) and CO(2) are reported. Nascent energy profiles for scattered CO(2) (00(0)0, J = 2-64) were measured using high-resolution transient IR absorption spectroscopy at lambda = 2.7 microm. The data are combined with earlier data for CO(2) (J = 58-80) to yield the full state-resolved distribution of scattered CO(2) (00(0)0). The scattered CO(2) (00(0)0, J = 2-80) molecules have a biexponential rotational distribution with T(lowJ) = 437 +/- 50 K for J < 50 and T(highJ) = 1145 +/- 110 K for J > 50. Fitting the data with a two component exponential model yields CO(2) product distributions with T(rot) = 329 and 1241 K. The cooler distribution accounts for 78% of the scattered population and results from elastic or weakly inelastic collisions that induce very little rotational excitation in CO(2). The hotter distribution involves large changes in CO(2) rotational energy and accounts for 22% of collisions. The relative translational energy of the scattered molecules increases as a function of final CO(2) rotational state with T(rel) = 860 K for J = 2, 1650 K for J = 60, and 5500 K for J = 80. The total rate constant for appearance of scattered CO(2) (00(0)0) is k(app) = (4.8 +/- 1.4) x 10(-10) cm(3) molecule(-1) s(-1), which is approximately 85% of the Lennard-Jones collision rate. Population depletion measurements yield a collision rate of k(dep) = (5.7 +/- 1.8) x 10(-10) cm(3) molecule(-1) s(-1), which is in very good agreement with the Lennard-Jones collision rate. The full energy transfer probability distribution function P(DeltaE) for pyrazine(E)-CO(2) collisions is presented and compared to results from other studies. PMID- 20000658 TI - Chiral indium alkoxide complexes as initiators for the stereoselective ring opening polymerization of rac-lactide. AB - The indium complex InL(2)N'' has been prepared from the reaction of 2 equiv of ((t)Bu)(2)P(O)CH(2)CH((t)Bu)OH (HL) with InN''(3) (N'' = N(SiMe(3))(2)). This complex reacts with a further equivalent of 2,6-di-tert-butylphenol or HL to afford the adducts InL(2)(OAr) and InL(3), respectively. Confirmation that the anion L(-) exhibits "ligand self-recognition" in the formation of predominantly homochiral complexes RR-InL(2)N'' and SS-InL(2)N'' is obtained from (1)H and (31)P NMR spectroscopic data. However, the self-recognition is less effective at the indium cation, and mixtures of InL(3) complexes with different configurations are observed. Single-crystal X-ray diffraction data confirm the five-coordinate, distorted bipyramidal In center in InL(2)N'' and InL(2)(OAr) as anticipated. Selected crystals of InL(3) show two of the possible configurations: one is the fac-RRR-InL(3) complex, analogous to the lanthanide complexes LnL(3) reported previously (Ln = Y, Eu, Er, Yb); another is the alternative, homochiral mer form RRR'-InL(3). All three complexes are efficient single-component initiators for the ring-opening polymerization of rac-lactide over a wide range of temperatures and monomer-to-initiator ratios, exhibiting reasonable control over the synthesis of isotactic polylactide. Despite its poorly defined structure, InL(3) is the fastest initiator among the three complexes for the polymerization of rac lactide, and shows the best tacticity control. The polylactide samples have high molecular weights M(n,exp) (between 44 000 and 270 000 g/mol at completion) and narrow polydispersities (as low as 1.25 at completion). PMID- 20000659 TI - Asymmetric synthesis of the potent HIV-protease inhibitor, nelfinavir. AB - An asymmetric synthesis of nelfinavir is described starting from acrolein and (S) methyl phenyl sulfoxide. The key features include (a) stereoselective preparation of a beta-protected amino-gamma,delta-unsaturated sulfoxide by the reaction of an alpha-sulfinyl carbanion with an unsaturated t-butyl sulfinylimine, (b) stereoselective bromohydrin formation using the pendant sulfoxide group as an intramolecular nucleophile, and (c) use of commercially or readily prepared inexpensive starting materials. PMID- 20000660 TI - Synthesis of the oxepinochromone natural products ptaeroxylin (desoxykarenin), ptaeroxylinol, and eranthin. AB - An improved synthesis of the oxepinochromone ptaeroxylin is reported, together with the syntheses of the related natural products ptaeroxylinol and eranthin. Ptaeroxylin and ptaeroxylinol were obtained from the chromenone noreugenin by selective reaction of the 7-hydroxyl group, allylation of the 5-hydroxyl, followed by Claisen rearrangement under microwave conditions with concomitant deprotection of the 7-hydroxyl. Alkylation of the 7-hydroxyl with the appropriate allyl bromide provides a precursor for ring-closing metathesis to deliver the oxepinochromone ring system. Eranthin was obtained by a similar strategy involving Claisen rearrangement to transfer an allyl group from the C-7 hydroxyl of noreugenin to C-8 regioselectively. PMID- 20000661 TI - Unprecedented 4- and 6-connected 2D coordination networks based on 4(4)-subnet tectons, showing unusual supramolecular motifs of rotaxane and helix. AB - Two unprecedented 2D coordination polymers with 4- and 6-connected topological nets, arising from the different linkages of two adjacent 4(4) layers, were prepared from Co(II), 1,3-bis(4-pyridyl)propane, and different isophthalate tectons. PMID- 20000663 TI - Critical size for the observation of quantum confinement in optically excited gold clusters. AB - We present a systematic study of optical properties of a series of hexanethiolate capped Au clusters of varying sizes using femtosecond transient absorption, time resolved fluorescence, and two-photon absorption cross-sectional measurements. An abrupt change in optical properties and their trends has been found at the 2.2 nm size. Displacively excited vibrations with a period of 450 fs have been detected in the transient absorption signal for smaller clusters < or = 2.2 nm. These results strongly suggest an emerging optical gap between the highest occupied and lowest unoccupied orbitals in the narrow size range at 2.2 nm. PMID- 20000662 TI - Monitoring the erosion of hydrolytically-degradable nanogels via multiangle light scattering coupled to asymmetrical flow field-flow fractionation. AB - We describe the synthesis and characterization of degradable nanogels that display bulk erosion under physiologic conditions (pH = 7.4, 37 degrees C). Erodible poly(N-isopropylmethacrylamide) nanogels were synthesized by copolymerization with N,O-(dimethacryloyl) hydroxylamine, a cross-linker previously used in the preparation of nontoxic and biodegradable bulk hydrogels. To monitor particle degradation, we employed multiangle light scattering and differential refractometry detection following asymmetrical flow field-flow fractionation. This approach allowed the detection of changes in nanogel molar mass and topology as a function of both temperature and pH. Particle erosion was evident from both an increase in nanogel swelling and a decrease in scattering intensity as a function of time. Following these analyses, the samples were recovered for subsequent characterization by direct particle tracking, which yields hydrodynamic size measurements and enables number density determination. Additionally, we confirmed the conservation of nanogel stimuli-responsivity through turbidity measurements. Thus, we have demonstrated the synthesis of degradable nanogels that erode under conditions and on time scales that are relevant for many drug delivery applications. The combined separation and light scattering detection method is demonstrated to be a versatile means to monitor erosion and should also find applicability in the characterization of other degradable particle constructs. PMID- 20000664 TI - High and selective CO2 uptake in a cobalt adeninate metal-organic framework exhibiting pyrimidine- and amino-decorated pores. AB - The synthesis and structure of Co(2)(ad)(2)(CO(2)CH(3))(2) x 2 DMF x 0.5 H(2)O (bio-MOF-11) is described. Pyrimidine and amino groups of adeninate (ad) decorate the pores of the framework. The porosity of this material was studied, and its CO(2) and H(2) adsorption properties were evaluated. bio-MOF-11 exhibits a high heat of adsorption for CO(2) (approximately 45 kJ/mol), a high CO(2) capacity (approximately 6 mmol/g, 273 K), and exceptional selectivity for CO(2) over N(2) at 273 K (81:1) and 298 K (75:1). PMID- 20000666 TI - Metal-metal multiple bonds in early/late heterobimetallics support unusual trigonal monopyramidal geometries at both Zr and Co. AB - Reduction of Zr/Co heterobimetallic complexes ICo(MesNP(i)Pr(2))(3)ZrCl (1) and ICo((i)PrNP(i)Pr(2))(3)ZrCl (2) with excess Na/Hg under N(2), followed by subsequent benzene extraction to remove coordinated Na halide salts, leads to neutral two-electron reduced, dinitrogen-bound complexes (THF)Zr(MesNP(i)Pr(2))(3)Co-N(2) (4) and Zr((i)PrNP(i)Pr(2))(3)Co-N(2) (5). Upon halide loss, a THF solvent molecule coordinates to the axial site of the Zr center in 4, while this axial site remains unoccupied in 5. X-ray crystallography reveals short Co-Zr distances in 4 and 5, indicative of metal-metal multiple bonding, and an unprecedented trigonal monopyramidal geometry about the Zr center in 5. Reduction of 4 under an Ar atmosphere (in the absence of N(2)) results in another unusual structure type: an unoccupied axial Co coordination site and a trigonal monopyramidal Co center in (THF)Zr(MesNP(i)Pr(2))(3)Co (6). X-ray crystallography reveals that, in the absence of coordinated N(2), the Co-Zr bond can attain full triple bond character with a Co-Zr distance of 2.14 A, the shortest M-M distance in an early/late heterobimetallic complex reported to date. To further assess the electronic structure and bonding in 4, 5, and 6, calculations were performed on these molecules using DFT and the results of these theoretical investigations will be discussed. PMID- 20000667 TI - Nanoscale chiral rod-like molecular triads assembled from achiral polyoxometalates. AB - The assembly of two achiral POMs of Lindqvist and Anderson through strong covalent linkages, using bifunctional TRIS with a rotatable C-N single bond, gives two nanoscale rod-like chiral molecular triads with C(2) symmetry, the chirality of which were confirmed by the single crystal X-ray diffraction analyses and solid CD spectroscopy measurements. The enantiopure crystals of these compounds have been obtained by spontaneous resolution upon crystallization in the absence of any chiral source. The present work opens a way to make chiral nanostructures from achiral polyoxoanions, which may have potential applications in asymmetric catalysis, NLO, and ferroelectric materials. PMID- 20000668 TI - 1,2,3-Triazole as a special "X-factor" in promoting hashmi phenol synthesis. AB - (31)P NMR experiments revealed rapid decomposition of the Ph(3)PAu(+)TfO(-) complex in the presence of the furan-yne, which resulted in poor reactivity as reported in the literature. Therefore, instead of tuning different ligands (PR(3)), the 1,2,3-triazole was applied as a special X-factor to stabilize the catalyst. The desired phenol products were prepared in excellent yields (1% cat. up to 95% yield) and chemoselectivity (>20:1). PMID- 20000669 TI - Allylcopper intermediates with N-heterocyclic carbene ligands: synthesis, structure, and catalysis. AB - Allylcopper intermediates with N-heterocyclic carbene ligands are synthesized by transmetalation of allylsiloxane reagents, and the crystal structures of allylcopper compounds are reported. The allylcopper transmetalation is utilized for catalytic aldehyde allylation, which is found to be facilitated by a trifluorosilane co-catalyst. PMID- 20000670 TI - Thermal decomposition mechanism of single-molecule precursors forming metal sulfide nanoparticles. AB - Several zinc alkyldithiocarbamates [Zn(S(2)CNR(2))(2)] were synthesized, and their thermal decomposition in the presence of alkylamines was studied in order to understand the formation of metal sulfide nanoparticles. The major intermediates and side products were isolated under various conditions and characterized by NMR spectroscopy and LC-MS. The analysis results showed that nucleophilic attack of the metal-coordinated amine on the most electron-deficient thiocarbonyl carbon of the alkyldithiocarbamate ligands at high temperature initiated the decomposition to generate thiourea, hydrogen sulfide, and solid metal sulfide nanoparticles. From the proposed decomposition mechanism, the role of amines in thermolysis was studied, and methods for synthesizing various metal sulfide nanoparticles were generalized. Formation of the desired products was confirmed by carrying out model experiments. PMID- 20000671 TI - Palladium-catalyzed decarboxylative benzylation of diphenylglycinate imines. AB - General reaction conditions for the Pd-catalyzed decarboxylative benzylation of benzyl diphenylglycinate imines are described. The overall procedure requires a simple catalyst/ligand combination to form a new Csp(3)-Csp(3) bond. Microwave irradiation greatly accelerated the transformation. Moreover, various heteroaromatic moieties are tolerated in both the imine and ester components. PMID- 20000672 TI - N,N-dialkylhydrazones in organic synthesis. From simple N,N-dimethylhydrazones to supported chiral auxiliaries. PMID- 20000673 TI - Nanoporous Au-Pt alloys as large strain electrochemical actuators. AB - Nanoporous Au-Pt alloys with pore- and ligament size down to few nanometers were fabricated by dealloying Ag-Au-Pt. Owing to the small structure size and large specific surface area, the surface stress and its variation give rise to significant stress and strain in the bulk of these materials. In fact, dilatometry experiments find electrochemical actuation with large reversible strain amplitude. The linear strain reaches approximately 1.3% and strain energy density is up to 6.0 MJ/m(3). The associated stresses may approach the elastic limit of the alloy. PMID- 20000674 TI - Transformation of Triclosan by Fe(III)-saturated montmorillonite. AB - Abiotic transformation of triclosan (TCS) was investigated by incubating TCS with Fe(III)- and Na-montmorillonite at 40% relative humidity and room temperature for up to 100 days. The TCS transformation products were characterized using LC/MS, GC/MS, and computational modeling and quantified using HPLC/UV and GC/MS. Within 1-5 days, depending on the initial TCS concentrations, about 55% of the TCS was rapidly transformed in the presence of Fe(III)-montmorillonite, producing 2,4 dichlorophenol, 3-chlorophenol, 2,4-dichlorophenol dimer, chlorophenoxy phenols, and TCS dimers and trimers. Computational modeling based on density functional theory confirmed the formation of four TCS dimer conformers and six TCS trimer conformers. The TCS phenoxy radicals, produced by Fe(III) oxidation of TCS, react with other TCS molecules to form TCS dimers. The TCS trimers were formed by attachment of TCS dimer phenoxy radicals, produced by Fe(III) oxidation of TCS dimers, with TCS molecules. Significantly smaller quantities of TCS transformation products were detected in the reactions with Na-montmorillonite compared to the reactions with Fe(III)-montmorillonite. Formation of a significant amount of 2,4-dichlorophenol, especially in reaction with Fe(III) montmorillonite, may have negative impact on the environment because of its toxicity. However, mineral-facilitated TCS polymerization may reduce its mobility and bioavailability in soils. PMID- 20000675 TI - Selective detection of airborne asbestos fibers using protein-based fluorescent probes. AB - Fluorescence microscopy (FM) is one of the most important analytical tools in modern life sciences, sufficiently sensitive to allow observation of single molecules. Here we describe the first application of the FM technique for the detection of inorganic environmental pollutants-airborne asbestos fibers that can cause asbestosis, mesothelioma, and lung cancer. In order to assess FM capabilities for detecting and counting asbestos fibers, we screened E. coli lysate for proteins that bind to amphibole asbestos. In combination with the previously discovered E. coli protein DksA (Kuroda et al., Biotechnol. Bioeng. 2008, 99, 285-289) that can specifically bind to chrysotile, the newly identified GatZ protein was used for selective and highly sensitive detection of two different asbestos types. Our novel FM-based method overcomes a number of limitations of the commonly used phase-contrast microscopy (PCM) method, offering a convenient alternative to PCM for airborne asbestos monitoring. PMID- 20000676 TI - EPR characterization and reactivity of surface-localized inorganic radicals and radical ions. PMID- 20000677 TI - Influence of the method of reagent addition on dichloroacetonitrile formation during chloramination. AB - Formation of dichloroacetonitrile (DCAN) in natural waters was evaluated for different disinfection scenarios, including application of free chlorine, preformed monochloramine or dichloramine, or formation of chloramines in situ by addition of free chlorine and ammonia in either order. Formation of DCAN was highest during free chlorination. Regardless of the order of ammonia or chlorine addition, DCAN formation was consistently higher over 1-2 day contact times when chloramines were formed in situ than when preformed chloramines were applied. During in situ chloramine formation, organic amine precursors effectively competed with ammonia to react with free chlorine, forming organic dichloramine intermediates to nitrile formation. Combined with previous research indicating that application of preformed monochloramine reduced nitrosamine formation, the results indicate that application of preformed monochloramine could provide an inexpensive alternative for chloraminating utilities to significantly reduce the exposure to DCAN and nitrosamines for consumers located within 1-2 days of water travel time from the treatment plant. This technique would be applicable in situations where chloramination is used alone (e.g., chlorination of non nitrified secondary wastewater effluents during municipal wastewater recycling), or combined with primary disinfectants other than free chlorine (e.g., ozonation, chlorine dioxide, or ultraviolet light). PMID- 20000678 TI - Development of a multistrain bacterial bioreporter platform for the monitoring of hydrocarbon contaminants in marine environments. AB - Petroleum hydrocarbons are common contaminants in marine and freshwater aquatic habitats, often occurring as a result of oil spillage. Rapid and reliable on-site tools for measuring the bioavailable hydrocarbon fractions, i.e., those that are most likely to cause toxic effects or are available for biodegradation, would assist in assessing potential ecological damage and following the progress of cleanup operations. Here we examined the suitability of a set of different rapid bioassays (2-3 h) using bacteria expressing the LuxAB luciferase to measure the presence of short-chain linear alkanes, monoaromatic and polyaromatic compounds, biphenyls, and DNA-damaging agents in seawater after a laboratory-scale oil spill. Five independent spills of 20 mL of NSO-1 crude oil with 2 L of seawater (North Sea or Mediterranean Sea) were carried out in 5 L glass flasks for periods of up to 10 days. Bioassays readily detected ephemeral concentrations of short chain alkanes and BTEX (i.e., benzene, toluene, ethylbenzene, and xylenes) in the seawater within minutes to hours after the spill, increasing to a maximum of up to 80 muM within 6-24 h, after which they decreased to low or undetectable levels. The strong decrease in short-chain alkanes and BTEX may have been due to their volatilization or biodegradation, which was supported by changes in the microbial community composition. Two- and three-ring PAHs appeared in the seawater phase after 24 h with a concentration up to 1 muM naphthalene equivalents and remained above 0.5 muM for the duration of the experiment. DNA damage-sensitive bioreporters did not produce any signal with the oil-spilled aqueous-phase samples, whereas bioassays for (hydroxy)biphenyls showed occasional responses. Chemical analysis for alkanes and PAHs in contaminated seawater samples supported the bioassay data, but did not show the typical ephemeral peaks observed with the bioassays. We conclude that bacterium-based bioassays can be a suitable alternative for rapid on-site quantitative measurement of hydrocarbons in seawater. PMID- 20000679 TI - Dual-action lipophilic iminosugar improves glycemic control in obese rodents by reduction of visceral glycosphingolipids and buffering of carbohydrate assimilation. AB - The lipophilic iminosugar N-[5-(adamantan-1-ylmethoxy)pentyl]-1-deoxynojirimycin (2, AMP-DNM) potently controls hyperglycemia in obese rodent models of insulin resistance. The reduction of visceral glycosphingolipids by 2 is thought to underlie its beneficial action. It cannot, however, be excluded that concomitant inhibition of intestinal glycosidases and associated buffering of carbohydrate assimilation add to this. To firmly establish the mode of action of 2, we developed a panel of lipophilic iminosugars varying in configuration at C-4/C-5 and N-substitution of the iminosugar. From these we identified the l-ido derivative of 2, l-ido-AMP-DNM (4), as a selective inhibitor of glycosphingolipid synthesis. Compound 4 lowered visceral glycosphingolipids in ob/ob mice and ZDF rats on a par with 2. In contrast to 2, 4 did not inhibit sucrase activity or sucrose assimilation. Treatment with 4 was significantly less effective in reducing blood glucose and HbA1c. We conclude that the combination of reduction of glycosphingolipids in tissue and buffering of carbohydrate assimilation by 2 produces a superior glucose homeostasis. PMID- 20000680 TI - Comparative analysis of halonitromethane and trihalomethane formation and speciation in drinking water: the effects of disinfectants, pH, bromide, and nitrite. AB - A bench-scale study was performed to investigate formation and speciation of halonitromethanes (HNMs) in raw and treated waters obtained from a drinking water treatment plant. HNM species were measured after chlorination, ozonation chlorination, chloramination, and ozonation-chloramination, and compared with trihalomethanes (THMs). Pre-ozonation before chlorination resulted in enhanced HNM formation, producing trihalogenated HNMs as major species. Formation of THMs showed very a different formation pattern from HNM formation such that it was much higher in the raw than the treated water, and decreased after pre-ozonation. These findings indicated that precursors and formation pathways for HNMs and THMs are not the same. Increases in pH and bromide concentrations increased HNM and THM formation during ozonation-chlorination and THM formation during chlorination. The bromide effect shifted the formation of HNMs and THMs toward brominated species, with its impact being greater in the treated than raw water. On the other hand, there was no pH or bromide effect on HNM formation during chlorination. The presence of nitrite increased HNM formation under both chlorination and ozonation-chlorination conditions, but it had no influence on THM formation. HNM formation during ozonation-chloramination remained about 1 microg/L level even at the highest bromide and nitrite concentrations. Monochloramination alone did not form any measurable HNMs. The results indicated that the use of chloramine can be an effective way to minimize HNM formation at typical drinking water treatment conditions. PMID- 20000681 TI - Biogeochemical redox processes and their impact on contaminant dynamics. AB - Life and element cycling on Earth is directly related to electron transfer (or redox) reactions. An understanding of biogeochemical redox processes is crucial for predicting and protecting environmental health and can provide new opportunities for engineered remediation strategies. Energy can be released and stored by means of redox reactions via the oxidation of labile organic carbon or inorganic compounds (electron donors) by microorganisms coupled to the reduction of electron acceptors including humic substances, iron-bearing minerals, transition metals, metalloids, and actinides. Environmental redox processes play key roles in the formation and dissolution of mineral phases. Redox cycling of naturally occurring trace elements and their host minerals often controls the release or sequestration of inorganic contaminants. Redox processes control the chemical speciation, bioavailability, toxicity, and mobility of many major and trace elements including Fe, Mn, C, P, N, S, Cr, Cu, Co, As, Sb, Se, Hg, Tc, and U. Redox-active humic substances and mineral surfaces can catalyze the redox transformation and degradation of organic contaminants. In this review article, we highlight recent advances in our understanding of biogeochemical redox processes and their impact on contaminant fate and transport, including future research needs. PMID- 20000682 TI - Well-architectured poly(dimethylsiloxane)-containing copolymers obtained by radical chemistry. PMID- 20000683 TI - Well-defined regions of the Plasmodium falciparum reticulocyte binding protein homologue 4 mediate interaction with red blood cell membrane. AB - Two widely studied parasite protein families are considered attractive targets for developing a fully effective antimalarial vaccine: the erythrocyte binding antigen (EBA) family defining a sialic acid-dependent invasion pathway, and reticulocyte-binding homologue (RH) proteins associated with sialic acid independent red blood cell (RBC) invasion. In this study, the micronemal invasive PfRH4 protein was finely mapped using 20-mer-long synthetic peptides spanning the entire protein length to identify protein regions that establish high affinity interactions with human RBCs. Twenty conserved, mainly alpha-helical high activity binding peptides (HABPs) with nanomolar dissociation constants and recognizing 32, 25, 22, and 20 kDa RBC membrane molecules in a chymotrypsin and/or trypsin-sensitive manner were identified in this protein. Anti-PfRH4 rabbit sera and PfRH4 HABPs inhibited merozoite invasion in vitro, therefore suggesting the implication of these HABPs in Plasmodium falciparum invasion and supporting their inclusion in further structural and immunological studies to design potential components of a minimal subunit-based, multiantigenic, chemically synthesized antimalarial vaccine. PMID- 20000684 TI - Genetic engineering of a bifunctional IgG fusion protein with iduronate-2 sulfatase. AB - Iduronate-2-sulfatase (IDS) is a lysosomal sulfatase that prevents the accumulation within the brain of glycosoaminoglycans. However, IDS does not cross the blood-brain barrier (BBB). To enable BBB transport, human IDS, minus its signal peptide, was fused to the carboxyl terminus of the heavy chain of a chimeric monoclonal antibody (mAb) to the human insulin receptor (HIR). The HIRMAb crosses the BBB on the endogenous insulin receptor and acts as a molecular Trojan horse to ferry the IDS into brain. The HIRMAb-IDS fusion protein was expressed in COS cells and purified with protein A affinity chromatography. The size of the fusion heavy chain, as measured with Western blotting and antibodies to either human IDS or human IgG, was increased about 80 kDa, relative to the size of the heavy chain of the parent HIRMAb. The HIRMAb-IDS fusion protein retained high-affinity binding for the HIR. The IDS enzyme specific activity of the fusion protein was 51 +/- 7 nmol/h per microgram of protein, which is comparable to the enzyme activity of recombinant IDS. The fusion protein was taken up by human fibroblasts, and the accumulation of glycosoaminoglycans in fibroblasts null for the sulfatase was decreased 84% by treatment with the fusion protein. The HIRMAb-IDS fusion protein is a bifunctional IgG-sulfatase fusion protein, which has been specifically engineered for targeted drug delivery across the human BBB. PMID- 20000685 TI - Synthesis of 4-methyl-1,2,3-thiadiazole derivatives via Ugi reaction and their biological activities. AB - The Ugi reaction is a green and rapid one-pot reaction for lead derivation. To develop novel candidate pesticides with diverse biological activities, two series of 4-methyl-1,2,3-thiadiazole derivatives containing active substructures of 3 chloro-4-methylphenyl or 3-fluoro-4-methylphenyl, respectively, were rationally designed and synthesized via Ugi reaction according to the principle of combinations of bioactive substructures. All of the structures of newly synthesized compounds were confirmed by proton nuclear magnetic resonance and high-resolution mass spectrometry. Biological activities of the target compounds including fungicide activity, antivirus activity in vitro and in vivo, and systemic acquired resistance were evaluated systematically. The results indicated that derivatives containing 3-(trifluoromethyl)phenyl and 2-methylphenyl possessed a potential wide spectrum of fungicidal activity. Derivatives containing 3-(trifluoromethyl)phenyl and 4-hydroxyphenyl possessed good potential direct antivirus activities against tobacco mosaic virus (TMV) in vitro, and the replacement of Cl atom by F atom improved their direct inhibition activities against TMV in vitro. Derivatives containing phenyl, 2-(trifluoromethyl)phenyl, 3 (trifluoromethyl)phenyl, 3-nitrophenyl, 4-nitrophenyl, 2-methylphenyl, and 4 hydroxyphenyl possessed good potential bioactivities in vivo including protection, inactivation, curative, and induction activities against TMV. These studies indicate that the newly synthesized 4-methyl-1,2,3-thiadiazole derivatives possessed good potential bioactivities, and a combination of bioactive substructures via Ugi reaction was an effective way to find bioactive compounds for novel pesticide development. PMID- 20000686 TI - Design, synthesis, and herbicidal activities of novel 2-cyanoacrylates containing isoxazole moieties. AB - A series of novel 2-cyanoacrylates containing an isoxazole moiety were designed and synthesized. Their structures were characterized by (1)H NMR and elemental analysis (or high-resolution mass spectrometry). Their herbicidal activities against four species were evaluated, and the results indicated that some of the title compounds showed excellent herbicidal activities against rape and amaranth pigweed in postemergence treatment even at a dose of 75 g/ha. PMID- 20000688 TI - A single glutamate residue controls the oligomerization, function, and stability of the aquaglyceroporin GlpF. AB - Like many other alpha-helical membrane proteins, the monomeric Escherichia coli aquaglyceroporin GlpF associates within cellular membranes and forms higher-order oligomeric structures. A potential impact of the oligomeric state on the protein function remains enigmatic. We have analyzed the role of residues W42 and E43 in the oligomerization of the E. coli GlpF protein in vitro and in vivo. In contrast to W42, the polar glutamate residue at position 43 appears to be critical for oligomerization. While other polar residues can substitute for the function of E43, replacement of E43 with alanine results in a greatly reduced GlpF oligomerization propensity. The reduced interaction propensity of GlpF E43A correlates with an impaired in vivo function as well as a decreased in vivo stability. Therefore, E43 is critical for the proper oligomerization of GlpF, and protein oligomerization appears to be crucial for the channel function as well as for the in vivo stability of the protein. PMID- 20000691 TI - Acetone precipitation of proteins and the modification of peptides. AB - Acetone precipitation is a common method for precipitation and concentration of proteins. We show here that a trace amount of residual acetone in the precipitated protein, can, after proteolysis, lead to selective modification of peptides predominantly those in which a glycine residue is the second amino acid, probably generating a relatively stable derivative that, under gas phase conditions, generates a y(1) ion of the same mass as proline. This modification is detectable by either MALDI-ToF or ESI-ion trap mass spectrometry and under normal sample preparation conditions is incomplete. The derivatization occurs in the condensed phase and is sufficiently stable that the modified peptide can elute on reversed phase chromatography at a different time to the unmodified peptide. Acetone precipitation is such a commonly used procedure in protein sample preparation for proteomics that some caution may be warranted. A significant number of peptides (about 5% of a typical proteome) meet the requirements for this reaction and could, therefore, change the outcome of studies. PMID- 20000689 TI - Large scale production of vesicles by hollow fiber extrusion: a novel method for generating polymersome encapsulated hemoglobin dispersions. AB - Vesicles, which include both liposomes and polymersomes (polymer vesicles), are being developed as therapeutic drug carriers. In this study, we present a fully scalable low pressure extrusion methodology for preparing vesicles. Vesicles were generated by continuous extrusion through a 200 nm pore diameter hollow fiber (HF) membrane. The first half of this study describes a method for generating empty polymersomes composed of different molecular weight amphiphilic poly(butadiene-b-ethylene oxide) (PBD-b-PEO) diblock copolymers on a large scale (50-100 mL) using a HF membrane. Monodisperse empty polymersomes were formed with particle diameters slightly less than 200 nm, which were close to the rated 200 nm pore size of the HF membrane. The second half of this study describes the successful encapsulation of hemoglobin (Hb) inside the aqueous core of polymersomes using the HF extrusion methodology. We demonstrate that polymersome encapsulated hemoglobin (PEH) particles formed by this technique had similar oxygen affinity, cooperativity coefficient, and methemoglobin (metHb) level compared to PEH particles formed by the 1 mL volume small scale manual extrusion method. Most notably, Hb encapsulation inside the polymer vesicles formed by the HF extrusion method increased 2-fold compared to the manual extrusion method. This work is important, since it will enable facile scale-up of homogeneous vesicle dispersions that are typically required for preclinical and clinical studies as well as industrial use. PMID- 20000692 TI - "Double click" reaction on 7-deazaguanine DNA: synthesis and excimer fluorescence of nucleosides and oligonucleotides with branched side chains decorated with proximal pyrenes. AB - The 7-tripropargylamine-7-deaza-2'-deoxyguanosine (2) containing two terminal triple bonds in the side chain was synthesized by the Sonogashira cross-coupling reaction from the corresponding 7-iodo nucleoside 1b. This was protected at the 2 amino group with an iso-butyryl residue, affording the protected intermediate 5. Then, compound 5 was converted to the 5'-O-DMT derivative 6, which on phosphitylation afforded the phosphoramidite 7. This was employed in solid-phase synthesis of a series of oligonucleotides. T(m) measurements demonstrate that a covalently attached tripropargylamine side chain increases duplex stability. Both terminal triple bonds of nucleoside 2 and corresponding oligonucleotides were functionalized by the Cu(I)-mediated 1,3-dipolar cycloaddition "double click reaction" with 1-azidomethyl pyrene 3, decorating the side chain with two proximal pyrenes. While the monomeric tripropargylamine nucleoside with two proximal pyrenes (4) shows strong excimer fluorescence, the ss-oligonucleotide containing 4 does not. This was also observed for ds-oligonucleotides when the complementary strand was unmodified. However, duplex DNA bearing pyrene residues in both strands exhibits strong excimer fluorescence when each strand contains two pyrene residues linked to the tripropargylamine moiety. This pyrene-pyrene interstrand interaction occurs when the pyrene modification sites of the duplex are separated by two base pairs which bring the fluorescent dyes in a proximal position. Molecular modeling indicates that only two out of four pyrene residues are interacting forming the exciplex while the other two do not communicate. PMID- 20000693 TI - A general and efficient synthesis of pyridin-2-yl C-ribonucleosides bearing diverse alkyl, aryl, amino, and carbamoyl groups in position 6. AB - An efficient and practical methodology of preparation of 6-substituted pyridin-2 yl C-ribonucleosides was developed. A one-pot two-step addition of 2-lithio-6 bromopyridine to TBS-protected ribonolactone followed by acetylation gave 1beta (6-bromopyridin-2-yl)-1-O-acetyl-2,3,5-tri-O-(tert-butyldimethylsilyl)-D ribofuranose in high yield. Its reduction with Et(3)SiH and BF(3) x Et(2)O afforded the desired TBS-protected 6-bromopyridine C-ribonucleoside as pure beta anomer in good overall yield of 63%. This intermediate was then subjected to a series of palladium catalyzed cross-coupling reactions, aminations and aminocarbonylations to give a series of protected 1beta-(6-alkyl-, 6-aryl-, 6 amino-, and 6-carbamoylpyridin-2-yl)-C-ribonucleosides. Deprotection of silylated nucleosides by Et(3)N x 3HF gave a series of title free C-ribonucleosides (12 examples). PMID- 20000694 TI - Acetylcholinesterase inhibitory pyridine alkaloids of the leaves of Senna multijuga. AB - Two unusual pyridine alkaloids, 7'-multijuguinone (1) and 12'-hydroxy-7' multijuguinone (2), were isolated from the leaves of Senna multijuga, together with the known flavonoid rutin. The structures of the new alkaloids were established on the basis of spectroscopic data interpretation. Compounds 1 and 2 exhibited moderate in vitro acetylcholinesterase (AChE) inhibitory activity, in comparison with the standard compound physostigmine. PMID- 20000695 TI - Quantification of (15)N-nitrate in urine with gas chromatography combustion isotope ratio mass spectrometry to estimate endogenous NO production. AB - The use of stable isotope labeled substrates and subsequent analysis of urinary nitrate, forms a noninvasive test for evaluation of the in vivo NO metabolism. The present paper describes a new method for simultaneous quantification of (15)N nitrate and total nitrate with gas chromatography combustion isotope ratio mass spectrometry (GC-C-IRMS). Nitrate, isolated from urine with a nitrate selective resin, was reduced to nitrite using copperized cadmium. Subsequently, Sudan I was formed by diazotation. Sudan II was added as internal standard, and both molecules were analyzed with GC-C-IRMS as tert-butyldimethylsilyl derivatives. The accuracy was determined during a recovery study of two different known nitrate concentrations and two (15)N-enrichments. A recovery of 101.6% and 103.9% for total nitrate and 107.6% and 91.2% for (15)N-nitrate was obtained, respectively. The validated method was applied on complete 72 h urine collections after intravenous administration of (15)N-nitrate and (15)N-arginine in humans. On average, 51.8% (47.0-71.0%) of administered (15)N-nitrate was excreted, while 0.68% (0.44-1.17%) of (15)N-arginine was metabolized to nitrate. In conclusion, this method can be used for accurate simultaneous determination of (15)N-nitrate and total nitrate concentrations in urine and can be applied in clinical studies for noninvasive evaluation of NO metabolism in vivo. PMID- 20000696 TI - Effects of architecture and surface chemistry of three-dimensionally ordered macroporous carbon solid contacts on performance of ion-selective electrodes. AB - The effects of the architecture and surface chemistry of three-dimensionally ordered macroporous (3DOM) carbon solid contacts on the properties of ion selective electrodes (ISEs) were examined. Infiltration of the plasticized poly(vinyl chloride) (PVC) membrane into the pores of the carbon created a large interfacial area between the sensing membrane and the solid contact, as shown by cryo-scanning electron microscopy (cryo-SEM) and elemental analysis. This large interfacial area, along with the high capacitance of the 3DOM carbon solid contacts (as determined by cyclic voltammetry, chronopotentiometry, and electrochemical impedance spectroscopy) results in an excellent long-term stability of the potentiometric response, with drifts as low as 11.7 muV/h. The comparison of 3DOM carbon solid contacts with an untemplated carbon solid contact shows that the pore structure is an essential feature for the excellent electrode performance. However, the surface chemistry of the 3DOM carbon cannot be ignored. While there is no evidence for an aqueous layer forming between the sensing membrane and unoxidized 3DOM carbon, electrodes based on oxidized 3DOM carbon exhibit potentiometric responses with the typical hysteresis indicative of a water layer. A comparison of the different techniques to characterize the solid contacts confirms that constant-current charge-discharge experiments offer an intriguing approach to assess the long-term stability of solid-contact ISEs but shows that their results need to be interpreted with care. PMID- 20000697 TI - Investigation of prothrombin time in human whole-blood samples with a quartz crystal biosensor. AB - Monitoring of blood coagulation and fibrinolysis is an important issue in treatment of patients with cardiovascular problems and in surgery when blood gets into contact with artificial surfaces. In this work a new method for measuring the coagulation time (prothrombin time, PT) of human whole-blood samples based on a quartz crystal microbalance (QCM) biosensor is presented. The 10 MHz sensors used in this work respond with a frequency shift to changes in viscosity during blood clot formation. For driving and for readout of the quartz, both a network analyzer and an oscillator circuit were utilized. The sensor surfaces were specifically coated with a thin polyethylene layer. We found that both frequency analysis methods are suitable to measure exact prothrombin times in a very good conformity with a mechanical coagulometer as a reference. The anticoagulant effect of heparin on the prothrombin time was exemplarily shown as well as the reverse effect of the heparin antagonist polybrene. The change of the viscoelastic properties during blood coagulation, reflected by the ratio of frequency and dissipation shifts, is discussed for different dilutions of the whole-blood samples. In conclusion, QCM is a distinguished biosensor technique to determine prothrombin time and to monitor heparin therapy in whole-blood samples. Due to the excellent potential of miniaturization and the availability of direct digital signals, the method is predestinated for incorporation and integration into other devices and is thus opening the field of application for inline coagulation diagnostic in extracorporeal blood circuits. PMID- 20000698 TI - Synthesis of chiral 3-substituted phthalides by a sequential organocatalytic enantioselective aldol-lactonization reaction. Three-step synthesis of (S)-(-)-3 butylphthalide. AB - The development of efficient methods for the facile construction of important molecular frameworks is an important goal in organic synthesis. Chiral 3 substituted phthalides are widely distributed in a large collection of natural products with broad, potent, and potentially path-pointing biological activities. In this investigation, we have uncovered an unprecedented organocatalytic asymmetric aldol-lactonization reaction of 2-formylbenzoic esters with ketones/aldehydes for convenient construction of the enantioenriched "privileged" scaffold. As a result of the sensitive nature of substrate structures of an organocatalytic enantioselective aldol reaction, after extensive optimization of reaction conditions, catalyst L-prolinamide alcohol IV is identified as the best promoter. Interestingly, it is found that in this reaction, addition of an acid additive PhCO(2)H can significantly enhance reaction efficiency with use of only as low as 2.5 mol % IV for the process. Moreover, due to the sensitivity of reaction conditions toward a sequential aldol-lactonization process without affecting enantioselectivity and racemization, it is essential to remove the catalyst for the subsequent facile lactonization reaction in the presence of K(2)CO(3). The aldol-lactonization processes serve as a powerful approach to the preparation of synthetically and biologically important 3-substituted phthalides with a high level of enantioselectivities. A 3-step catalytic asymmetric synthesis of the natural product of 3-butylphthalide is reported. PMID- 20000699 TI - Accurate MALDI-TOF/TOF sequencing of one-bead-one-compound peptide libraries with application to the identification of multiligand protein affinity agents using in situ click chemistry screening. AB - Combinatorial one-bead-one-compound (OBOC) peptide libraries are widely used for affinity screening, and the sequencing of peptides from hit beads is a key step in the process. For rapid sequencing, CNBr cleavage of the peptides from the beads, followed by de novo sequencing by MALDI-TOF/TOF, is explored. We report on a semiautomated sequencing algorithm and validate it through comparison against Edman degradation sequencing. The initial 44% sequencing success rate of the standard de novo sequencing software was improved to nearly 100%. The sequencing algorithm incorporates existing knowledge of amino acid chemistry and a new strategy for differentiating isobaric amino acids. We tested the algorithm by using MALDI-TOF/TOF to identify a peptide biligand affinity agent against the protein bovine carbonic anhydrase II, starting from comprehensive one-bead-one compound peptide libraries comprised of non-natural and artificial amino acid components and using the strategy of in situ click/OBOC library screening. PMID- 20000700 TI - Ruthenium-based heterocyclic carbene-coordinated olefin metathesis catalysts. PMID- 20000701 TI - Efficient alpha-(alkylthio)alkyl-type radical formation in (*)OH-induced oxidation of alpha-(methylthio)acetamide. AB - Pulse radiolysis with UV-vis/ESR detection and steady-state gamma-radiolysis, combined with chromatographic techniques, were used to investigate the detailed mechanism of the (*)OH-induced oxidation of alpha-(methylthio)acetamide (alpha MTA) in aqueous solution. The main pathway involves the formation of hydroxysulfuranyl radicals alpha-MTA-(>S(*)-OH) and alpha-(alkylthio)alkyl radicals H(3)C-S-(*)CH-C( horizontal lineO)-NH(2) (lambda(max) S(*) OH) radicals undergo efficient conversion to intermolecularly three-electron bonded dimeric radical cations of alpha-MTA-(>S thereforeS<)(+) (lambda(max) = 480 nm), especially for high alpha-MTA concentrations. In contrast, at low proton concentrations, alpha-MTA-(>S(*)-OH) radicals decompose via the elimination of water, formed through intramolecular hydrogen (attached to the nitrogen atom) transfer to the hydroxysulfuranyl moiety within a six-membered structure. This process leads to the formation of the imine radical H(3)C-S-CH(2)-C( horizontal lineO)(*)NH, which subsequently decays in three independent channels. The first decay channel begins with a beta-scission followed by hydrolysis and a subsequent Hofmann rearrangement. One of the end products of this first decay channel is CO(2), which was detected. The second decay channel involves an intramolecular hydrogen transfer from the deltaC carbon atom to the radical imine site producing the alpha-(alkylthio)alkyl radical H(2)C(*)-S-CH(2)-C( horizontal lineO)-NH(2). In the third decay channel there is a 1,3-hydrogen shift in the imine radical which forms the radical H(3)C-S-(*)CH-C( horizontal lineO)-NH(2). The presence of the amide group induces more complex radical chemistry that leads unexpectedly to the degradation of the CH(3)SCH(2)CONH(2) molecule into gaseous products, CO(2) and NH(3). These features of the mechanism of the (*)OH-induced oxidation of alpha-MTA are quite different from those seen in other organic sulfides in neutral solutions. PMID- 20000702 TI - Establishment of an evaluation model for human milk fat substitutes. AB - Fatty acid composition and distribution of human milk fat (HMF), from mothers over different lactating periods in Guangzhou, China, were analyzed. The universal characteristics were consistent with previously reported results although the fatty acid content was within a different range and dependent on the local population (low saturated fatty acid and high oleic acid for Guangdong mothers' milk fat). Based on the composition of the total and sn-2 fatty acids of mature milk fat, an efficient evaluation model was innovatively established by adopting the "deducting score" principle. The model showed good agreement between the scores and the degree of similarity by assessing 15 samples from different sources including four samples of HMF, eight samples of human milk fat substitutes (HMFSs) and infant formulas, and three samples of fats and oils. This study would allow for the devolvement of individual human milk fat substitutes with different and specific fatty acid compositions for local infants. PMID- 20000703 TI - Measurement of retention efficiency of filters against nanoparticles in liquids using an aerosolization technique. AB - An aerosolization technique has been developed to measure liquid-borne nanoparticles down to 30 nm and applied to evaluate retention efficiencies of liquid filters. This technique involves dispersing nanoparticle suspensions into air-borne form and measuring the size and concentration by a differential mobility analyzer coupled to a condensation particle counter. Polystyrene latex particles larger and smaller than 70 nm in diameter were dispersed by a constant output atomizer, COA, and an electrospray aerosol generator, ES, respectively, to avoid the interference from residue particles. With the ES, residue particles can be controlled to be less than 10 nm, allowing latex particles as small as 30 nm to be clearly distinguished from the size distribution measurements. Calibrations with 30, 50, 125, and 200 nm latex particles showed that liquid-borne and air borne particle concentrations are proportionally related. This provides an effective way to quantify liquid-borne particles as small as 30 nm, which cannot be analyzed by state-of-the-art liquid particle counters. An application of this technique is to evaluate the nanoparticle retention performance of liquid filters. Both 200 and 400 nm rated Nuclepore filters were challenged with latex particles of different sizes, and retention efficiency as a function of particle size was determined by comparing the particle concentrations upstream and downstream of the tested filters. The results are comparable with the nominal pore size stated by the manufacturer if sieving is the dominant filtration mechanism and demonstrate the feasibility of using the aerosolization technique to evaluate the retention efficiency of filters against nanoparticles in liquids. PMID- 20000705 TI - Orthogonal orientation control of carbon nanotube growth. AB - Carbon nanotubes (CNTs) have attracted attention for their remarkable electrical properties and have being explored as one of the best building blocks in nano electronics. A key challenge to realize such potential is the control of the nanotube growth directions. Even though both vertical growth and controlled horizontal growth of carbon nanotubes have been realized before, the growth of complex nanotube structures with both vertical and horizontal orientation control on the same substrate has never been achieved. Here, we report a method to grow three-dimensional (3D) complex nanotube structures made of vertical nanotube forests and horizontal nanotube arrays on a single substrate and from the same catalyst pattern by an orthogonally directed nanotube growth method using chemical vapor deposition (CVD). More importantly, such a capability represents a major advance in controlled growth of carbon nanotubes. It enables researchers to control the growth directions of nanotubes by simply changing the reaction conditions. The high degree of control represented in these experiments will surely make the fabrication of complex nanotube devices a possibility. PMID- 20000704 TI - An antibiotic-resistance enzyme from a deep-sea bacterium. AB - We describe herein a highly proficient class A beta-lactamase OIH-1 from the bacterium Oceanobacillus iheyensis, whose habitat is the sediment at a depth of 1050 m in the Pacific Ocean. The OIH-1 structure was solved by molecular replacement and refined at 1.25 A resolution. OIH-1 has evolved to be an extremely halotolerant beta-lactamase capable of hydrolyzing its substrates in the presence of NaCl at saturating concentration. Not only is this the most highly halotolerant bacterial enzyme structure known to date, it is also the highest resolution halophilic protein structure yet determined. Evolution of OIH 1 in the salinity of the ocean has resulted in a molecular surface that is coated with acidic residues, a marked difference from beta-lactamases of terrestrial sources. OIH-1 is the first example of an antibiotic-resistance enzyme that has evolved in the depths of the ocean in isolation from clinical selection and gives us an extraordinary glimpse into protein evolution under extreme conditions. It represents evidence for the existence of a reservoir of antibiotic-resistance enzymes in nature among microbial populations from deep oceanic sources. PMID- 20000706 TI - High acceptance of urine source separation in seven European countries: a review. AB - Urine source separation (NoMix-technology) is a promising innovation aiming at a resource-oriented, decentralized approach in urban water management. However, NoMix-technology has a sensitive end-point: people's bathrooms. NoMix-technology is increasingly applied in European pilot projects, but the success from a user point-of-view has rarely been systematically monitored. We aim at closing this gap. We review surveys on acceptance, including reuse of human urine as fertilizer, from 38 NoMix-projects in 7 Northern and Central European countries with 2700 respondents. Additionally, we identify explanatory variables with logistic regression of a representative Swiss library survey. NoMix-technology is well accepted; around 80% of users liked the idea, 75-85% were satisfied with design, hygiene, smell, and seating comfort of NoMix-toilets, 85% regarded urine fertilizers as good idea (50% of farmers), and 70% would purchase such food. However, 60% of users encountered problems; NoMix-toilets need further development. We found few differences among countries, but systematic differences between public and private settings, where people seem more critical. Information was positively correlated with acceptance, and, e.g., a good mood or environmentally friendly behavior. For future success of NoMix-projects, we recommend authorities follow an integral strategy. Lay people will then find the NoMix-concept appealing and support this promising bathroom innovation. PMID- 20000707 TI - Chiral lithium(I) binaphtholate salts for the enantioselective direct Mannich type reaction with a change of syn/anti and absolute stereochemistry. AB - A highly diastereo- and enantioselective direct Mannich-type reaction of aldimines with 1,3-dicarbonyl compounds using Li(I) BINOLate salts as effective Lewis acid-Bronsted base catalysts has been developed. Li(I) BINOLate salts offered high catalytic activity toward 1,3-dicarbonyl compounds such as diketone, ketoester, ketothioester, ketoamide, and ketolactone. The reactions proceeded at 78 degrees C within 1-2 h in the presence of 1-10 mol % catalyst, which showed a catalytic activity (turnover frequency = 284 h(-1)) quite unlike those of other previous catalysts. Anti products were selectively obtained from acyclic ketoesters without epimerization at an alpha-tertiary-carbon center, and these are valuable since previous catalysts often gave syn/anti mixtures or the stereochemistry has not yet been determined. PMID- 20000708 TI - User-loaded SlipChip for equipment-free multiplexed nanoliter-scale experiments. AB - This paper describes a microfluidic approach to perform multiplexed nanoliter scale experiments by combining a sample with multiple different reagents, each at multiple mixing ratios. This approach employs a user-loaded, equipment-free SlipChip. The mixing ratios, characterized by diluting a fluorescent dye, could be controlled by the volume of each of the combined wells. The SlipChip design was validated on an approximately 12 nL scale by screening the conditions for crystallization of glutaryl-CoA dehydrogenase from Burkholderia pseudomallei against 48 different reagents; each reagent was tested at 11 different mixing ratios, for a total of 528 crystallization trials. The total consumption of the protein sample was approximately 10 microL. Conditions for crystallization were successfully identified. The crystallization experiments were successfully scaled up in well plates using the conditions identified in the SlipChip. Crystals were characterized by X-ray diffraction and provided a protein structure in a different space group and at a higher resolution than the structure obtained by conventional methods. In this work, this user-loaded SlipChip has been shown to reliably handle fluids of diverse physicochemical properties, such as viscosities and surface tensions. Quantitative measurements of fluorescent intensities and high-resolution imaging were straighforward to perform in these glass SlipChips. Surface chemistry was controlled using fluorinated lubricating fluid, analogous to the fluorinated carrier fluid used in plug-based crystallization. Thus, we expect this approach to be valuable in a number of areas beyond protein crystallization, especially those areas where droplet-based microfluidic systems have demonstrated successes, including measurements of enzyme kinetics and blood coagulation, cell-based assays, and chemical reactions. PMID- 20000709 TI - Multiparameter screening on SlipChip used for nanoliter protein crystallization combining free interface diffusion and microbatch methods. AB - This paper describes two SlipChip-based approaches to protein crystallization: a SlipChip-based free interface diffusion (FID) method and a SlipChip-based composite method that simultaneously performs microbatch and FID crystallization methods in a single device. The FID SlipChip was designed to screen multiple reagents, each at multiple diffusion equilibration times, and was validated by screening conditions for crystallization of two proteins, enoyl-CoA hydratase from Mycobacterium tuberculosis and dihydrofolate reductase/thymidylate synthase from Babesia bovis, against 48 different reagents at five different equilibration times each, consuming 12 microL of each protein for a total of 480 experiments using three SlipChips. The composite SlipChip was designed to screen multiple reagents, each at multiple mixing ratios and multiple equilibration times, and was validated by screening conditions for crystallization of two proteins, enoyl CoA hydratase from Mycobacterium tuberculosis and dihydrofolate reductase/thymidylate synthase from Babesia bovis. To prevent cross-contamination while keeping the solution in the neck channels for FID stable, the plates of the SlipChip were etched with a pattern of nanowells. This nanopattern was used to increase the contact angle of aqueous solutions on the surface of the silanized glass. The composite SlipChip increased the number of successful crystallization conditions and identified more conditions for crystallization than separate FID and microbatch screenings. Crystallization experiments were scaled up in well plates using conditions identified during the SlipChip screenings, and X-ray diffraction data were obtained to yield the protein structure of dihydrofolate reductase/thymidylate synthase at 1.95 A resolution. This free-interface diffusion approach provides a convenient and high-throughput method of setting up gradients in microfluidic devices and may find additional applications in cell based assays. PMID- 20000710 TI - Probing molecular motion by double-quantum (13C,13C) solid-state NMR spectroscopy: application to ubiquitin. AB - We demonstrate the use of two-dimensional ((13)C,(13)C) double-quantum spectroscopy to detect molecular dynamics by solid-state NMR. Data collected on tyrosine-ethylester (TEE) are in line with previously determined ((1)H,(13)C) order parameters. Application of these experiments to microcrystalline ubiquitin reveals the presence of dynamics on millisecond or faster time scales and differences in local mobility depending on microcrystal preparation. In addition, solid-state NMR-based structure calculation indicates conformational variability of loop regions between different solid-phase ubiquitin preparations. Our data relate preparation-dependent changes observed in NMR spectral parameters such as chemical shifts and through-space correlations to differences in ubiquitin dynamics and conformation and suggest a prominent role of molecular mobility in microcrystalline ubiquitin. PMID- 20000712 TI - Contrasting magnetism of [Mn(III)(4)] and [Mn(II)(2)Mn(III)(2)] squares. AB - Two tetranuclear manganese distorted square-shaped clusters, [Mn(III)(4)(L1)(4)(mu(2)-OMe)(4)].2.5H(2)O (1) and [Mn(II)(2)Mn(III)(2)(L2)(4)(H(2)O)(2)](PF(6))(2).CHCl(3).CH(3)OH.1.5H(2)O (2) (H(2)L1 = 2-[3-(2-hydroxyphenyl)-1H-pyrazol-5-yl]-6-pyridinecarboxylic acid methyl ester; H(2)L2 = 2-[3-(2-hydroxyphenyl)-1H-pyrazol-5-yl]-6 pyridinecarboxylic acid ethyl ester), exhibit antiferromagnetic and ferromagnetic interactions between neighboring manganese ions, respectively. PMID- 20000713 TI - Tripodal 4-pyridyl-derived host ligands and their metallo-supramolecular chemistry: stella octangula and bowl-shaped assemblies. AB - The synthesis of five new cyclotriveratrylene derivatives with 4-pyridyl side arms is reported, along with the crystal structures of three of these. Three ligands with extended 4-pyridylphenyl side arms and a ligand derived from cyclotriphenolene have been shown to form [Pd(6)L(8)](12+) stella octangula assemblies using diffusion-ordered spectroscopy NMR and electrospray MS techniques. This confirms the generality of the stella octagula assembly, providing that the ligand arms show a degree of rigidity. The more flexible ether linked ligand tris(4-pyridylmethyl)cyclotriguaiacylene forms a smaller [Pd(3)L(4)](6+) bowl-shaped assembly in the solid state and in solution. The previously reported ligand tris(4-pyridylmethylamino)cyclotriguaiacylene forms a similar assembly in solution. PMID- 20000711 TI - Influence of the nitrogen donors on nonheme iron models of superoxide reductase: high-spin Fe(III)-OOR complexes. AB - A new five-coordinate, (N(4)S(thiolate))Fe(II) complex, containing tertiary amine donors, [Fe(II)(Me(4)[15]aneN(4))(SPh)]BPh(4) (2), was synthesized and structurally characterized as a model of the reduced active site of superoxide reductase (SOR). Reaction of 2 with tert-butyl hydroperoxide (tBuOOH) at -78 degrees C led to the generation of the alkylperoxo-iron(III) complex [Fe(III)(Me(4)[15]aneN(4))(SPh)(OOtBu)](+) (2a). The nonthiolate-ligated complex, [Fe(II)(Me(4)[15]aneN(4))(OTf)(2)] (3), was also reacted with tBuOOH and yielded the corresponding alkylperoxo complex [Fe(III)(Me(4)[15]aneN(4))(OTf)(OOtBu)](+) (3a) at an elevated temperature of -23 degrees C. These species were characterized by low-temperature UV-vis, EPR, and resonance Raman spectroscopies. Complexes 2a and 3a exhibit distinctly different spectroscopic signatures than the analogous alkylperoxo complexes [Fe(III)([15]aneN(4))(SAr)(OOR)](+), which contain secondary amine donors. Importantly, alkylation at nitrogen leads to a change from low-spin (S = 1/2) to high-spin (S = 5/2) of the iron(III) center. The resonance Raman data reveal that this change in spin state has a large effect on the nu(Fe-O) and nu(O-O) vibrations, and a comparison between 2a and the nonthiolate-ligated complex 3a shows that axial ligation has an additional significant impact on these vibrations. To our knowledge this study is the first in which the influence of a ligand trans to a peroxo moiety has been evaluated for a structurally equivalent pair of high-spin/low-spin peroxo-iron(III) complexes. The implications of spin state and thiolate ligation are discussed with regard to the functioning of SOR. PMID- 20000714 TI - Kinetics and products of the OH radical-initiated reaction of 1,4-butanediol and rate constants for the reactions of OH radicals with 4-hydroxybutanal and 3 hydroxypropanal. AB - Rate constants for the gas-phase reactions of the OH radical with 1,4 butanediol,4-hydroxybutanal, and 3-hydroxypropanalwere measured at 298 +/- 2 K and atmospheric pressure using a relative rate technique and with 4 hydroxybutanal and 3-hydroxypropanal being formed in situ from the OH + 1,4 butanediol reaction, and were (in units of 10(-11) cm(3) molecule(-1) s(-1)) 3.67 +/- 0.31, 3.0(-1.0)(+1.5), and 3.5(-1.0)(+1.3), respectively, with the latter being a lower limit because of the possibility of second-generation formation of 3-hydroxypropanal from OH + 4-hydroxybutanal. The measured formation yields of 4 hydroxybutanal and 3-hydroxypropanal from the OH + 1,4-butanediol reaction in the presence of NO were 58 +/- 19% and 55 +/- 12%, respectively. In addition to the formation of 4-hydroxybutanal and 3-hydroxypropanal, gas chromatography-mass spectrometric analyses of reacted OH + 1,4-butanediol reactionsshowedformation of glycolaldehyde [HOCH(2)CHO], malonaldehyde [HC(O)CH2CHO], and 1,4-butanedial [HC(O)CH(2)CH(2)CHO]. Glycolaldehyde, malonaldehyde, and 1,4-butanedial are expected products of the reaction of OH radicals with 4-hydroxybutanal, and glycolaldehyde and malonaldehyde are expected products of the reaction of OH radicals with 3-hydroxypropanal. Reaction mechanisms are presented and discussed. PMID- 20000715 TI - Heterometallic 3d-4f polyoxometalate derived from the weakley-type dimeric structure. AB - Polyanion [{Ce(H(2)O)(2)}(2)Mn(2)(B-alpha-GeW(9)O(34))(2)](8-) (1) constitutes the first example of a heterometallic 3d-4f cluster related to the Weakley-type dimeric structure, and it contains an unprecedented Ce(III)(2)Mn(III)(2)O(20) rhomblike moiety displaying dominant Ce(III)-Mn(III) ferromagnetic interactions. PMID- 20000717 TI - Liesegang rings engineered from charged nanoparticles. AB - Functionalized nanoparticles (NPs) serve as building blocks of self-organizing chemical patterns comprising periodic zones of nanoparticle precipitation. In contrast to ions, which underlie most pattern-forming chemical systems and whose properties cannot be readily modified, NPs allow for flexible adjustment of particle charges and/or material properties. In particular, changes in the particle charges control the precipitation behavior and ultimately the morphologies of the emerging patterns. The phenomenon of NP-based periodic precipitation is explained by reaction-diffusion modeling and can be used for the fractionation of NPs of different sizes. PMID- 20000716 TI - A comparative study on conformation and ligand binding of the neuronal uncoupling proteins. AB - Mitochondrial uncoupling proteins of the nervous system (UCPs 2, 4, and 5) have potential roles in the function and protection of the central nervous system (CNS). In the absence of structural information, conformations of the hexahistidine-tagged versions of all five human UCPs in liposomes were investigated for the first time, using far- and near-UV CD and fluorescence spectroscopy. Highly pure UCPs 1-5 were reconstituted in detergents and stable small unilamellar vesicles, appropriate for spectroscopic studies. All UCPs formed dominantly helical conformations in negatively charged phospholipid vesicles (palmitoyloleoylphosphatidylcholine/palmitoyloleoylphosphatidylglycerol, 7:3 molar ratio). UCPs 2 and 5 exhibited comparable helical conformations with possible association in lipid bilayers, whereas UCP4 had a different helical profile that can be related to its less associated form. Interaction of reconstituted UCPs with GDP and GTP, inhibitors of the prototypic UCP1, was detected by near-UV CD and fluorescence spectroscopy, utilizing the sensitivity of these techniques to microenvironments around Trp residues close to the nucleotide binding site. Binding of UCP4 to purine nucleotides was also different from other UCPs. Binding of fatty acids, activators of proton transport in UCPs, to UCPs could not be unambiguously detected, implying a nonbinding conformation/orientation of the proteoliposomes. Interaction of CoA with UCPs was comparable to nucleotide binding, suggesting a possible binding of this molecule at the nucleotide binding site. Despite dissimilar primary sequences, neuronal UCPs share common structural and functional properties with UCPs 1 and 3, supporting a common physiological role in addition to their specific roles in the CNS. PMID- 20000718 TI - Highly fluorescent BODIPY dyes modulated with spirofluorene moieties. AB - A new type of structurally rigid BODIPY dye having spirofluorene moieties has been synthesized. These structurally constrained BODIPY dyes with spirofluorene moieties exhibit an intense bathochromic fluorescence. The solvent dependence of fluorescence ON/OFF switching of a BODIPY dye having an amino moiety was observed. PMID- 20000719 TI - Thiol-modified chitosan sulfate nanoparticles for protection and release of basic fibroblast growth factor. AB - A series of chitosan (CS) derivatives, the 6-O-carboxymethylchitosan (6-O-CC), 2 N sulfated 6-O-carboxymethylchitosan (N-SOCC) and the 2-N and 3,6-O sulfated 6-O carboxymethyl chitosan (N,O-SOCC) were synthesized in this study. The chemical structures and the degrees of substituted carboxymethyl and sulfate groups of the synthesized compounds were respectively determined by FT-IR spectra and elemental analysis. N,O-SOCC displayed the highest protective efficiency for basic fibroblast growth factor (bFGF) as examined by the L929 fibroblast culture test and docking simulation. N,O-SOCC-4-thio-butylamidine (TBA) conjugates prepared by modification of N,O-SOCC with 2-iminothiolane were in situ cross-linkable. The degrees of thiol substitution of the 2-iminothiolane modified N,O-SOCC polymers were determined to be in the ranges of 45.9 +/- 3.7 and 415.6 +/- 12.5 micromol SH/g SOCC by quantifying the amount of thiol groups on the thiolated polymers with Ellman's reagent. The 2-iminothiolane modified N,O-SOCC and CS complex could be used for preparing nanoparticles by a polyelectrolyte self-assembly method, and the release of bFGF from the nanoparticles was successfully controlled. L929 fibroblast culture tests showed that the thiol modified N,O-SOCC/CS nanoparticles could effectively protect bFGF from inactivation over a 120 h period. The results of this study suggest that the thiol modified N,O-SOCC/CS nanoparticles may be useful as novel materials for specific delivery of bFGF with mitogenic activity. PMID- 20000720 TI - Making mixtures to solve structures: structural elucidation via combinatorial synthesis. AB - A domino Horner-Wadsworth-Emmons olefination strategy has been used to prepare homologous series of (polyen)ones, and through combinatorial elaboration, corresponding families of highly branched hydrocarbons. Gas chromatography-mass spectrometry of the mixtures has enabled the rapid and unambiguous identification of several highly branched alkanes of geochemical importance. This is the first example of the use of combinatorial synthesis for the elucidation of structural connectivity. PMID- 20000721 TI - Growth of crystalline Gd2O3 thin films with a high-quality interface on Si(100) by low-temperature H2O-assisted atomic layer deposition. AB - This work documents the first example of deposition of high-quality Gd(2)O(3) thin films in a surface-controlled, self-limiting manner by a water-based atomic layer deposition (ALD) process using the engineered homoleptic gadolinium guanidinate precursor [Gd(DPDMG)(3)]. The potential of this class of compound is demonstrated in terms of a true ALD process, exhibiting pronounced growth rates, a high-quality interface between the film and the substrate without the need for any additional surface treatment prior to the film deposition, and most importantly, encouraging electrical properties. PMID- 20000722 TI - Fluoroolefins as peptide mimetics. 2. A computational study of the conformational ramifications of peptide bond replacement. AB - The design of peptide mimetic compounds is greatly facilitated by the identification of functionalities that can act as peptide replacements. The fluoroalkene moiety has recently been employed for that purpose. The purpose of this work is to examine the conformational ramifications of replacing peptide bonds with fluoroalkene moieties, thus generating peptidomimetics. The alanine dipeptide analogue (ADA) was chosen as a model compound. Three peptidomimetic systems were investigated including one generated by replacement of both peptide bonds of ADA, designated as DFA, and those generated by the single replacement of the C-terminal peptide bond and N-terminal peptide bond, designated as CFA and NFA, respectively. Conformations for all three systems were generated by exhaustive Monte Carlo searching. Relative conformational energies were calculated at the MP2/aug-cc-pVTZ/MP2/aug-cc-pVDZ (for DFA), MP2/-aug-cc pVTZ//MP2/6-311+G(d,p), B3LYP/6-31+G(d)//B3LYP/6-31+G(d), and MMFF levels of theory. Aqueous phase conformational preferences were determined through calculations making use of continuum hydration models. The results indicate that replacement of both peptide bonds of ADA generates a peptidomimetic with conformational preferences where extended conformations are favored and the conformational profile is relatively insensitive to the nature of the surrounding medium. This is in contrast to ADA where the conformational preferences depend highly on the surrounding medium and where folded conformations with intramolecular hydrogen bonds are important in the absence of an interacting solvent. CFA and NFA are found to exhibit conformational preferences that do in some ways more closely resemble those of the alanine dipeptide analogue. This is particularly true in the case of NFA where interactions between the NH and CF groups are reminiscent of the intramolecular hydrogen bonding possible in ADA. PMID- 20000723 TI - Protonation of type-1 Cu bound histidines: a quantum chemical study. AB - The protonation of the solvent-exposed histidine ligands of the type-1 Cu sites in five proteins, Thiobacillus ferrooxidans rusticyanin, Pseudomonas aeruginosa azurin, fern plastocyanin, Alcaligenes faecalis pseudoazurin, and Paracoccus versutus amicyanin, were studied with quantum chemical methods and conductorlike polarizable continuum model (CPCM). Active site model molecules consisting of approximately 140 atoms were extracted from X-ray crystal structures and optimized with the homogeneous CPCM/B3LYP/6-31G* method with some atoms fixed. More accurate solvation effects were obtained using a recently developed heterogeneous CPCM method to describe the protein matrix and aqueous solvation of the model molecules. In the heterogeneous CPCM method different effective dielectric constants, 4, 10, and 78.39, were used for different portions of the surfaces encapsulating the active site model molecules. It is found that the two conformations of the protonated histidine, imidazolium unflipped and flipped, show similar energies in the model molecules of these five proteins. The calculated pK(a) values are comparable to experimental values. According to the calculations, the main determinants of the pK(a) values are local interactions contained in the model molecules and aqueous solvation effects, as well as protein matrix polarization. PMID- 20000724 TI - Improved hydrophilic interaction chromatography LC/MS of heparinoids using a chip with postcolumn makeup flow. AB - Heparan sulfate (HS) and heparin are linear, heterogeneous carbohydrates of the glycosaminoglycan (GAG) family that are modified by N-acetylation, N-sulfation, O sulfation, and uronic acid epimerization. HS interacts with growth factors in the extracellular matrix, thereby modulating signaling pathways that govern cell growth, development, differentiation, proliferation, and adhesion. High performance liquid chromatography (HPLC)-chip-based hydrophilic interaction liquid chromatography/mass spectrometry has emerged as a method for analyzing the domain structure of GAGs. However, analysis of highly sulfated GAG structures decasaccharide or larger in size has been limited by spray instability in the negative-ion mode. This report demonstrates that addition of postcolumn makeup flow to the amide-HPLC-chip configuration permits robust and reproducible analysis of extended GAG domains (up to degree of polymerization 18) from HS and heparin. This platform provides quantitative information regarding the oligosaccharide profile, degree of sulfation, and nonreducing chain termini. It is expected that this technology will enable quantitative, comparative glycomics profiling of extended GAG oligosaccharide domains of functional interest. PMID- 20000725 TI - Controlled synthesis of iron oxide nanoparticles over a wide size range. AB - We report on the effect of using decanoic acid as capping ligand on the synthesis of iron oxide nanoparticles by thermal decomposition of an organic iron precursor in organic medium. This procedure allowed us to control the particle size within 5 nm and about 30 nm by modifying the precursor-to-capping ligand ratio in a systematic fashion and to further expand the particle size range up to about 50 nm by adjusting the final synthesis temperature. The nanoparticles also showed high saturation magnetization of about 80-83 emu/g at low temperature, almost size-independent and close to the value for the bulk counterpart. Decanoic acid coated nanoparticles were transferred to water by using tetramethylammonium hydroxide, which allowed further coating with silica in a tetraethyl orthosilicate solution. Consequently, these iron oxide nanoparticles are tunable in size and highly magnetic, and they could become suitable candidates for various biomedical applications such as contrast agents for magnetic resonance imaging and magnetic carriers for drug delivery. PMID- 20000726 TI - Quantitative analysis of transverse bacterial migration induced by chemotaxis in a packed column with structured physical heterogeneity. AB - A two-dimensional mathematical model was developed to simulate transport phenomena of chemotactic bacteria in a sand-packed column designed with structured physical heterogeneity in the presence of a localized chemical source. In contrast to mathematical models in previous research work, in which bacteria were typically treated as immobile colloids, this model incorporated a convective like chemotaxis term to represent chemotactic migration. Consistency between experimental observation and model prediction supported the assertions that (1) dispersion-induced microbial transfer between adjacent conductive zones occurred at the interface and had little influence on bacterial transport in the bulk flow of the permeable layers and (2) the enhanced transverse bacterial migration in chemotactic experiments relative to nonchemotactic controls was mainly due to directed migration toward the chemical source zone. On the basis of parameter sensitivity analysis, chemotactic parameters determined in bulk aqueous fluid were adequate to predict the microbial transport in our intermediate-scale porous media system. Additionally, the analysis of adsorption coefficient values supported the observation of a previous study that microbial deposition to the surface of porous media might be decreased under the effect of chemoattractant gradients. By quantitatively describing bacterial transport and distribution in a heterogeneous system, this mathematical model serves to advance our understanding of chemotaxis and motility effects in granular media systems and provides insights for modeling microbial transport in in situ microbial processes. PMID- 20000727 TI - On the Maxwell-Stefan approach to diffusion: a general resolution in the transient regime for one-dimensional systems. AB - The diffusion process in a multicomponent system can be formulated in a general form by the generalized Maxwell-Stefan equations. This formulation is able to describe the diffusion process in different systems, such as, for instance, bulk diffusion (in the gas, liquid, and solid phase) and diffusion in microporous materials (membranes, zeolites, nanotubes, etc.). The Maxwell-Stefan equations can be solved analytically (only in special cases) or by numerical approaches. Different numerical strategies have been previously presented, but the number of diffusing species is normally restricted, with only few exceptions, to three in bulk diffusion and to two in microporous systems, unless simplifications of the Maxwell-Stefan equations are considered. In the literature, a large effort has been devoted to the derivation of the analytic expression of the elements of the Fick-like diffusion matrix and therefore to the symbolic inversion of a square matrix with dimensions n x n (n being the number of independent components). This step, which can be easily performed for n = 2 and remains reasonable for n = 3, becomes rapidly very complex in problems with a large number of components. This paper addresses the problem of the numerical resolution of the Maxwell-Stefan equations in the transient regime for a one-dimensional system with a generic number of components, avoiding the definition of the analytic expression of the elements of the Fick-like diffusion matrix. To this aim, two approaches have been implemented in a computational code; the first is the simple finite difference second-order accurate in time Crank-Nicolson scheme for which the full mathematical derivation and the relevant final equations are reported. The second is based on the more accurate backward differentiation formulas, BDF, or Gear's method (Shampine, L. F. ; Gear, C. W. SIAM Rev. 1979, 21, 1.), as implemented in the Livermore solver for ordinary differential equations, LSODE (Hindmarsh, A. C. Serial Fortran Solvers for ODE Initial Value Problems, Technical Report; https://computation.llnl.gov/casc/odepack/odepack_ home.html (2006).). Both methods have been applied to a series of specific problems, such as bulk diffusion of acetone and methanol through stagnant air, uptake of two components on a microporous material in a model system, and permeation across a microporous membrane in model systems, both with the aim to validate the method and to add new information to the comprehension of the peculiar behavior of these systems. The approach is validated by comparison with different published results and with analytic expressions for the steady-state concentration profiles or fluxes in particular systems. The possibility to treat a generic number of components (the limitation being essentially the computational power) is also tested, and results are reported on the permeation of a five component mixture through a membrane in a model system. It is worth noticing that the algorithm here reported can be applied also to the Fick formulation of the diffusion problem with concentration dependent diffusion coefficients. PMID- 20000728 TI - Detailed kinetic Monte Carlo simulations of graphene-edge growth. AB - A new detailed chemical-kinetic Monte Carlo model of graphene-edge growth is presented. The model employs a fine-grained approach to chemically resolved species, allows for incorporation of five-member rings into growing structures, and links the stochastic kinetic steps to a geometry optimization, thereby properly accounting for curving of molecular structures. The evolving morphology is greatly affected by the rates of key reactions and hence by surface-site steric environment and gas-phase species concentrations. The evolving graphene morphology and growth rates seemingly reach "asymptotic" behavior, independent of the initial substrate. Most noteworthy, growing layers become significantly curved. The curvature occurs regardless of initial substrate at both 1500 and 2000 K with higher curvature occurring at the lower temperature. More intriguing is the observation that, at 2000 K, transition from planar to curved growth does not commence immediately but occurs at some later time, seemingly when the growing graphene reaches a size significantly larger than coronene. No curvature is produced in numerical simulations at 2500 K, indicating that high-energy environments cause the five-member-ring to be less stable, thus preventing them from forming. PMID- 20000729 TI - Synthesis of azide-alkyne fragments for "click" chemical applications. Part 2. Formation of oligomers from orthogonally protected chiral trialkylsilylhomopropargyl azides and homopropargyl alcohols. AB - A small library of chiral, beta(3)-substituted homopropargyl alcohols and chiral beta(3)-substituted trimethylsilylhomopropargyl azides were generated starting from natural l-amino acids. The free alkynes and azides were then coupled, using a Huisgen 1,3-dipolar cycloaddition, to provide chiral oligomeric 1,4 disubstituted-1,2,3-triazoles as potential peptidomimetic compounds. The work is an extension to the previous synthesis of racemic, orthogonally protected 1,4 disubstituted-1,2,3-triazoles from the corresponding alpha-substituted propargyl alcohols and alpha-substituted trialkylsilylpropargyl azides. PMID- 20000730 TI - Quantitative Correlation between the protein primary sequences and secondary structures in spider dragline silks. AB - Synthetic spider silk holds great potential for use in various applications spanning medical uses to ultra lightweight armor; however, producing synthetic fibers with mechanical properties comparable to natural spider silk has eluded the scientific community. Natural dragline spider silks are commonly made from proteins that contain highly repetitive amino acid motifs, adopting an array of secondary structures. Before further advances can be made in the production of synthetic fibers based on spider silk proteins, it is imperative to know the percentage of each amino acid in the protein that forms a specific secondary structure. Linking these percentages to the primary amino acid sequence of the protein will establish a structural foundation for synthetic silk. In this study, nuclear magnetic resonance (NMR) techniques are used to quantify the percentage of Ala, Gly, and Ser that form both beta-sheet and helical secondary structures. The fraction of these three amino acids and their secondary structure are quantitatively correlated to the primary amino acid sequence for the proteins that comprise major and minor ampullate silk from the Nephila clavipes spider providing a blueprint for synthetic spider silks. PMID- 20000731 TI - A tripod molecular tip for single molecule ligand-receptor force spectroscopy by AFM. AB - Tripod-shaped molecules were designed for chemical modification of the surface of probes used for atomic force microscopy (AFM). These chemically functionalized tips were used for chemical force spectroscopy (CFS) measurements of the ligand protein receptor interaction in a biotin-NeutrAvidin model system. We demonstrate that by using this unique tripodal system, we can achieve significantly lower density of ligand on the AFM tip apex, which is optimal for true single molecule measurements. Furthermore, the molecular tripods form highly stable bonds to the AFM probes, leading to more robust and reproducible unbinding force data, thereby addressing one of the challenges in CFS studies. Histogram analysis of the hundreds of collected unbinding forces showed a specific distribution with a peak force maximum at approximately 165 pN, in good agreement with the previously reported data of single rupture events of biotin-avidin. We compared these molecular tripod tips with a molecular monopod. The results showed that the molecular tripods are more robust for repeated measurements. The distinct biotin avidin force maximum was not observed in the control experiments. This indicated that the force distribution observed for molecular tripods corresponds to the specific rupture force between biotin and avidin. The improved robustness of molecular tripods for CFS will provide benefits in other ligand-receptor unbinding studies, including those of transmembrane receptor systems, which require high resolution, sensitivity, and reproducibility in force spectroscopy measurements. PMID- 20000732 TI - Degradation of 1,2,3-trichloropropane (TCP): hydrolysis, elimination, and reduction by iron and zinc. AB - 1,2,3-Trichloropropane (TCP) is an emerging contaminant because of increased recognition of its occurrence in groundwater, potential carcinogenicity, and resistance to natural attenuation. The physical and chemical properties of TCP make it difficult to remediate, with all conventional options being relatively slow or inefficient. Treatments that result in alkaline conditions (e.g., permeable reactive barriers containing zerovalent iron) favor base-catalyzed hydrolysis of TCP, but high temperature (e.g., conditions of in situ thermal remediation) is necessary for this reaction to be significant. Common reductants (sulfide, ferrous iron adsorbed to iron oxides, and most forms of construction grade or nano-Fe(0)) produce insignificant rates of reductive dechlorination of TCP. Quantifiable rates of TCP reduction were obtained with several types of activated nano-Fe(0), but the surface area normalized rate contants (k(SA)) for these reactions were lower than is generally considered useful for in situ remediation applications (10(-4) L m(-2) h(-1)). Much faster rates of degradation of TCP were obtained with granular Zn(0), (k(SA) = 10(-3) - 10(-2) L m(-2) h(-1)) and potentially problematic dechlorination intermediates (1,2- or 1,3 dichloropropane, 3-chloro-1-propene) were not detected. The advantages of Zn(0) over Fe(0) are somewhat peculiar to TCP and may suggest a practical application for Zn(0) even though it has not found favor for remediation of contamination with other chlorinated solvents. PMID- 20000733 TI - TiCl4-promoted Baylis-Hillman reaction: mechanistic rationale toward product distribution and stereoselectivity. AB - The mechanism of TiCl(4)-promoted Baylis-Hillman reaction between methyl vinyl ketone (MVK) and acetaldehyde, in the absence of any base, is studied using the mPW1K density functional theory. The study focuses on several mechanistic intricacies as well as selectivity issues at each step of the reaction. The minimum energy pathway for this reaction involves three major steps such as a chloride transfer resulting in a chloro-enolate, titanium-mediated aldol reaction, and elimination of HCl or HOTiCl(3). Both s-cis and s-trans conformers of MVK are considered along with various modes of chloride transfer involving different complexes between TiCl(4), aldehyde, and MVK. Chloride transfer is found to be kinetically more favored for s-cis-MVK than for s-trans-MVK. The diastereoselectivity in the next step, i.e., Ti-mediated aldol reaction between the enolate and aldehyde, is found to be dependent on the geometry of the enolate, wherein anti and syn BH products are predicted for Z and E enolates, respectively. An interesting secondary orbital interaction between the oxygen atoms of the enolate and aldehyde moieties in the transition states for the C-C bond formation is identified as one of the contributing factors toward the predicted diastereoselectivity in the formation of the alpha-chloromethyl aldol product (P2). It has earlier been reported that under different experimental conditions, any of the three products such as (i) a normal BH product (P1), (ii) 2-(chloromethyl)vinyl ketones (P3), and (iii) alpha-chloro methyl aldol could be generated (Scheme 1 ). The present study offers valuable insights toward rationalizing the observed product distribution as well as diastereoselectivity in TiCl(4)-promoted BH reaction under base-free conditions. The computed energetics indicate that when MVK is employed as the Michael acceptor, the formation of 2-(choromethyl)vinyl ketone is the preferred product rather than the corresponding normal BH product, consistent with the known experimental reports. PMID- 20000734 TI - Direct space methods for powder X-ray diffraction for guest-host materials: applications to cage occupancies and guest distributions in clathrate hydrates. AB - Structural determination of crystalline powders, especially those of complex materials, is not a trivial task. For non-stoichiometric guest-host materials, the difficulty lies in how to determine dynamical disorder and partial cage occupancies of the guest molecules without other supporting information or constraints. Here, we show how direct space methods combined with Rietveld analysis can be applied to a class of host-guest materials, in this case the clathrate hydrates. We report crystal structures in the three important hydrate crystal classes, sI, sII, and sH, for the guests CO(2), C(2)H(6), C(3)H(8), and methylcyclohexane + CH(4). The results obtained for powder samples are found to be in good agreement with the experimental data from single crystal X-ray diffraction and (13)C solid-state NMR spectroscopy. This method is also used to determine the guest disorder and cage occupancies of neohexane and tert-butyl methyl ether binary hydrates with CH(4) in the structure H clathrate hydrates. The results are found to be in good agreement with the results from the (13)C solid-state NMR and molecular dynamics simulations. It is demonstrated that the ab initio crystal structure determination methodology reported here is able to determine absolute cage occupancies and the dynamical disorder of guest molecules in clathrate hydrates from powdered crystalline samples. PMID- 20000735 TI - Through the "gatekeeper door": exploiting the active kinase conformation. PMID- 20000736 TI - Gold-catalyzed cyclization of alkynylaziridines as an efficient approach toward functionalized N-phth pyrroles. AB - An efficient access to N-phth pyrrroles via gold-catalyzed cycloisomerization of N-phth alkynylaziridines has been described. Functionalized pyrroles including pyrrole-2-carboxylates or 2-pyrrolyl ketone are easily constructed in generally good yields by this method. The resulting pyrroles can be further converted to N amino pyrrole or 2-acyl pyrrole, which are important synthetic intermediates for amplification of molecular complexity. PMID- 20000737 TI - Isotopic and elemental data for tracing the origin of European olive oils. AB - H, C, and O stable isotope ratios and the elemental profile of 267 olive oils and 314 surface waters collected from 8 European sites are presented and discussed. The aim of the study was to investigate if olive oils produced in areas with different climatic and geological characteristics could be discriminated on the basis of isotopic and elemental data. The stable isotope ratios of H, C, and O of olive oils and the ratios of H and O of the relevant surface waters correlated to the climatic (mainly temperature) and geographical (mainly latitude and distance from the coast) characteristics of the provenance sites. It was possible to characterize the geological origin of the olive oils by using the content of 14 elements (Mg, K, Ca, V, Mn, Zn, Rb, Sr, Cs, La, Ce, Sm, Eu, U). By combining the 3 isotopic ratios with the 14 elements and applying a multivariate discriminant analysis, a good discrimination between olive oils from 8 European sites was achieved, with 95% of the samples correctly classified into the production site. PMID- 20000739 TI - Chemical and geometrical criteria for the release of elastomeric 1D nanoarrays from porous nanotemplates. AB - Releasing 1D nanoarrays from nanotemplates is a significant challenge for the integration of mechanically soft materials in a variety of newly emerging technological areas. To fabricate nanoarrays without defects, the combined effects of the surface energy and the geometric features of the nanotemplate should be considered. A previously reported approach based on the correlation between the adhesion energy and the real contact area was not satisfactory to describe the rupture conditions of the nanofibers while they were being peeled off from the porous template. Here we demonstrate that the aspect ratio rather than the contact area of the nanoporous template is the key factor determining the upper limit of the pore length of the nanotemplate with respect to the rupture of the nanoarray during separation. We propose that the value of alpha(c)*, which is calculated with a simple expression in which the adhesion energy is multiplied by the aspect ratio, can be used as an excellent criterion for the fabrication of 1D nanoarrays without defects with a simple peel-off processes. Our approach opens up new applications for unconventional lithographic techniques, such as soft lithography, imprint lithography, and others. PMID- 20000740 TI - Chelating Langmuir-Blodgett film: a new versatile chemiluminescent sensing layer for biosensor applications. AB - The present study reports the achievement of a new chemiluminescent sensing layer able to simultaneously (i) play an active role on ligand immobilization and (ii) serve as a catalyst in detection processes for label-free biosensor applications. This new type of active Langmuir-Blodgett (LB) monolayer has been designed by using a chelating lipid (Ni-NTA-DOGS). Thanks to the chelated metallic cation, this peculiar lipid exhibits luminol chemiluminescence catalysis properties in the presence of hydrogen peroxide. Upon biomolecule interaction through imidazole ring chelation (mediated by the metallic cation bound to the lipid headgroup), the chemiluminescent signal can be modulated. The first chemiluminescent signal acquisition experiments have shown a strong and homogeneous signal of the chelating layer. Upon histamine interactions, a histidine derivative used as a marker of fresh food quality, we succeeded in obtaining as a proof of concept a chemiluminescent signal variation without any derivatization of the target molecule. This signal variation was shown to be directly correlated to the histamine concentration with a limit of detection of 2 microg/mL. PMID- 20000738 TI - Proteomic dissection of cell type-specific H2AX-interacting protein complex associated with hepatocellular carcinoma. AB - The replacement histone variant H2AX senses DNA double-strand breaks (DSBs) and recruits characteristic sets of proteins at its phosphorylated (gamma-H2AX) foci for concurrent DNA repair. We reasoned that the H2AX interaction network, or interactome, formed in the tumor-associated DNA DSB environment such as in hepatocellular carcinoma (HCC) cells, where preneoplastic lesions frequently occur, is indicative of HCC pathogenic status. By using an in vivo dual-tagging quantitative proteomic method, we identified 102 H2AX-specific interacting partners in HCC cells that stably expressed FLAG-tagged H2AX at close to the endogenous level. Using bioinformatics tools for data-dependent network analysis, we further found binary relationships among these interactors in defined pathway modules, implicating H2AX in a multifunctional role of coordinating a variety of biological pathways involved in DNA damage recognition and DNA repair, apoptosis, nucleic acid metabolism, Ca(2+)-binding signaling, cell cycle, etc. Furthermore, our observations suggest that these pathways interconnect through key pathway components or H2AX interactors. The physiological accuracy of our quantitative proteomic approach in determining H2AX-specific interactors was evaluated by both coimmunoprecipitation/ immunoblotting and confocal colocalization experiments performed on HCC cells. Due to their involvement in diverse functions, the H2AX interactors involved in different pathway modules, such as Poly(ADP-ribose) polymerase 1, 14-3-3 zeta, coflin 1, and peflin 1, were examined for their relative H2AX binding affinities in paired hepatocytes and HCC cells. Treatment with the DSB-inducing agent bleomycin enhanced binding of these proteins to H2AX, suggesting an active role of H2AX in coordinating the functional pathways of each protein in DNA damage recognition and repair. PMID- 20000741 TI - Structural insights into the dual activities of the nerve agent degrading organophosphate anhydrolase/prolidase. AB - The organophosphate acid anhydrolase (OPAA) is a member of a class of bimetalloenzymes that hydrolyze a variety of toxic acetylcholinesterase inhibiting organophosphorus compounds, including fluorine-containing chemical nerve agents. It also belongs to a family of prolidases, with significant activity against various Xaa-Pro dipeptides. Here we report the X-ray structure determination of the native OPAA (58 kDa mass) from Alteromonas sp. strain JD6.5 and its cocrystal with the inhibitor mipafox [N,N' diisopropyldiamidofluorophosphate (DDFP)], a close analogue of the nerve agent organophosphate substrate diisopropyl fluorophosphate (DFP). The OPAA structure is composed of two domains, amino and carboxy domains, with the latter exhibiting a "pita bread" architecture and harboring the active site with the binuclear Mn(2+) ions. The native OPAA structure revealed unexpectedly the presence of a well-defined nonproteinaceous density in the active site whose identity could not be definitively established but is suggestive of a bound glycolate, which is isosteric with a glycine (Xaa) product. All three glycolate oxygens coordinate the two Mn(2+) atoms. DDFP or more likely its hydrolysis product, N,N' diisopropyldiamidophosphate (DDP), is present in the cocrystal structure and bound by coordinating the binuclear metals and forming hydrogen bonds and nonpolar interactions with active site residues. An unusual common feature of the binding of the two ligands is the involvement of only one oxygen atom of the glycolate carboxylate and the product DDP tetrahedral phosphate in bridging the two Mn(2+) ions. Both structures provide new understanding of ligand recognition and the prolidase and organophosphorus hydrolase catalytic activities of OPAA. PMID- 20000743 TI - Prioritization of candidate protein biomarkers from an in vitro model system of breast tumor progression toward clinical verification. AB - The use of in vitro cell culture model systems has revealed many potential mediators and candidate biomarkers of various disease phenotypes. To be of clinical utility, the expression of these candidates must be assessed in patient samples such as tissue, urine or blood. However, typical "omic" experiments may produce candidates in such large numbers that it is usually impossible to test all of these in clinical samples. Here, we present a proteomic approach to discover and prioritize candidate biomarkers that are more likely to be found in serum. Using a combination of experimental and in silico approaches, we have demonstrated this approach using an isogenic cell culture model of breast cancer invasion. Differential proteomics (2D-DIGE) was used to discover a number of candidate biomarkers and a subset of these were identified as "extracellular". We tested the validity of this approach by screening serum from breast cancer patients for these candidates and then verified the presence of several of these "extracellular" proteins. This approach provides a pragmatic approach to prioritizing candidates that may be most suitable for downstream assays such as multiple reaction monitoring. PMID- 20000742 TI - The essential Escherichia coli apolipoprotein N-acyltransferase (Lnt) exists as an extracytoplasmic thioester acyl-enzyme intermediate. AB - Escherichia coli apolipoprotein N-acyltransferase (Lnt) transfers an acyl group from sn-1-glycerophospholipid to the free alpha-amino group of the N-terminal cysteine of apolipoproteins, resulting in mature triacylated lipoprotein. Here we report that the Lnt reaction proceeds through an acyl-enzyme intermediate in which a palmitoyl group forms a thioester bond with the thiol of the active site residue C387 that was cleaved by neutral hydroxylamine. Lnt(C387S) also formed a fatty acyl intermediate that was resistant to neutral hydroxylamine treatment, consistent with formation of an oxygen-ester linkage. Lnt(C387A) did not form an acyl-enzyme intermediate and, like Lnt(C387S), did not have any detectable Lnt activity, indicating that acylation cannot occur at other positions in the catalytic domain. The existence of this thioacyl-enzyme intermediate allowed us to determine whether essential residues in the catalytic domain of Lnt affect the first step of the reaction, the formation of the acyl-enzyme intermediate, or the second step in which the acyl chain is transferred to the apolipoprotein substrate. In the catalytic triad, E267 is required for the formation of the acyl enzyme intermediate, indicating its role in enhancing the nucleophilicity of C387. E343 is also involved in the first step but is not in close proximity to the active site. W237, Y388, and E389 play a role in the second step of the reaction since acyl-Lnt is formed but N-acylation does not occur. The data presented allow discrimination between the functions of essential Lnt residues in catalytic activity and substrate recognition. PMID- 20000744 TI - Surface plasmon mediated strong exciton-photon coupling in semiconductor nanocrystals. AB - We present an experimental demonstration of strong coupling between a surface plasmon propagating on a planar silver thin film and the lowest excited state of CdSe nanocrystals. Attenuated total reflection measurements demonstrate the formation of plasmon-exciton mixed states, characterized by a Rabi splitting of approximately 112 meV at room temperature. Such a coherent interaction has the potential for the development of nonlinear plasmonic devices, and furthermore, this system is akin to those studied in cavity quantum electrodynamics, thus offering the possibility to study the regime of strong light-matter coupling in semiconductor nanocrystals under easily accessible experimental conditions. PMID- 20000745 TI - Spin of semiconductor quantum dots under hydrostatic pressure. AB - Spin coherence dynamics of semiconductor quantum dots under hydrostatic pressure has been investigated by combining the ultrafast optical orientation method with the diamond-anvil cell technique. Spin confined within quantum dots is observed to be robust up to several gigapascals, while electron and exciton Lande g factors show novel bistable characteristics prior to the first-order structural transition. This observation is attributed to the existence of a theoretically predicted metastable intermediate state at the nanoscale, for which there has been no previous experimental support. The results also reveal pressure enhanced fundamental exchange interactions for large-sized quantum dots with sizable anisotropy. These findings shed insight into underlying mechanisms of long debated nanoscale solid-state transformations in semiconductors and are also crucial for the development of future quantum information processing and manipulation based on spin qubits of quantum dots. PMID- 20000746 TI - Inter-regional variability in environmental availability of fungicide derived copper in vineyard soils: an Australian case study. AB - This study determined the environmental availability of copper (Cu) in Australian vineyard soils contaminated with fungicide derived Cu residues, and investigated the soil characteristics correlated with differences in Cu availability between regions. Concentrations of 0.01 M calcium chloride extractable Cu, measured in surface soils collected from 98 vineyards in 10 different grape-growing regions of Australia, ranged from <0.1 to 0.94 mg/kg and accounted for 0.10-1.03% of the total Cu concentrations in the soils. Differences in the calcium chloride extractable Cu concentrations were related to the total Cu concentration and soil properties, including pH, clay, exchangeable K, silt, and calcium carbonate. The information generated from this study may prove useful in devising strategies to reduce the availability and toxicity of Cu in agricultural soils. PMID- 20000747 TI - Identification of flavonoid and phenolic antioxidants in black currants, blueberries, raspberries, red currants, and cranberries. AB - The antioxidant capacity (AOC) of black currant, blueberry, raspberry, red currant, and cranberry extracts was determined using the FRAP assay. In addition, the vitamin C content of the berries was determined and phenolic and polyphenolic compounds in the extracts were analyze by reversed-phase HPLC-PDA-MS(3) and by reversed-phase HPLC-PDA with an online antioxidant detection system. A complex spectrum of anthocyanins was the major contributor to the AOC of black currants and blueberries, whereas the lower AOC of red currants and cranberries was due mainly to a reduced anthocyanin content. Raspberries also had a lower anthocyanin content than black currants and blueberries, but there was only a slight decline in the AOC because of the presence of the ellagitannins sanguin H-6 and lambertianin C, which were responsible for 58% of the HPLC-AOC of the berries. Vitamin C was responsible for 18-23% of the HPLC-AOC of black currants, red currants, and cranberries and for 11% of that of raspberries but did not contribute to the AOC of the blueberry extract that was examined. Seven percent of the HPLC-AOC of the cranberry extract was attributable to procyanidin dimers. However, the contribution of polymeric proanthocyanidins to the AOC of the five berries was not determined as when analyzed by reversed-phase HPLC these high molecular weight flavan-3-ols are either retained by the column or eluted as a broad unresolved band. PMID- 20000748 TI - Formation of {[HIPTN(3)N]Mo(III)H}(-) by heterolytic cleavage of H(2) as established by EPR and ENDOR spectroscopy. AB - MoN(2) (Mo = [(HIPTNCH(2)CH(2))(3)N]Mo, where HIPT = 3,5-(2,4,6-i Pr(3)C(6)H(2))(2)C(6)H(3)) is the first stage in the reduction of N(2) to NH(3) by Mo. Its reaction with dihydrogen in fluid solution yields "MoH(2)", a molybdenum-dihydrogen compound. In this report, we describe a comprehensive electron paramagnetic resonance (EPR) and (1/2)H/(14)N electron nuclear double resonance (ENDOR) study of the product of the reaction between MoN(2) and H(2) that is trapped in frozen solution, 1. EPR spectra of 1 show that it has a near axial g tensor, g = [2.086, 1.961, 1.947], with dramatically reduced g anisotropy relative to MoN(2). Analysis of the g values reveal that this anion has the Mo(III), [d(xz), d(yz)](3) orbital configuration, as proposed for the parent MoN(2) complex, and that it undergoes a strong pseudo-Jahn-Teller (PJT) distortion. Simulations of the 2D 35 GHz (1)H ENDOR pattern comprised of spectra taken at multiple fields across the EPR envelope (2 K) show that 1 is the [MoH]( ) anion. The 35 GHz Mims pulsed (2)H ENDOR spectra of 1 prepared with (2)H(2) show the corresponding (2)H(-) signal, with a substantial deuterium isotope effect in a(iso). Radiolytic reduction of a structural analogue, Mo(IV)H, at 77 K, confirms the assignment of 1. Analysis of the 2D (14)N ENDOR pattern for the ligand amine nitrogen further reveals the presence of a linear N(ax)-Mo-H(-) molecular axis that is parallel to the unique magnetic direction (g(1)). The ENDOR pattern of the three equatorial nitrogens is well-reproduced by a model in which the Mo-N(eq) plane has undergone a static, not dynamic, PJT distortion, leading to a range of hyperfine couplings for the three N(eq)'s. The finding of a nearly axial hyperfine coupling tensor for the terminal hydride bound Mo supports the earlier proposal that the two exchangeable hydrogenic species bound to the FeMo cofactor of the nitrogense turnover intermediate, which has accumulated four electrons/protons (E(4)), are hydrides that bridge two metal ions, not terminal hydrides. PMID- 20000750 TI - Microorganisms in dry polar snow are involved in the exchanges of reactive nitrogen species with the atmosphere. AB - The snowpack is a complex photochemical reactor that emits a wide variety of reactive molecules to the atmosphere. In particular, the photolysis of nitrate ions, NO(3)(-), produces NO, NO(2), and HONO, which affects the oxidative capacity of the atmosphere. We report measurements in the European High Arctic where we observed for the first time emissions of NO, NO(2), and HONO by the seasonal snowpack in winter, in the complete or near-complete absence of sunlight and in the absence of melting. We also detected unusually high concentrations of nitrite ions, NO(2)(-), in the snow. These results suggest that microbial activity in the snowpack is responsible for the observed emissions. Isotopic analysis of NO(2)(-) and NO(3)(-) in the snow confirm that these ions, at least in part, do not have an atmospheric origin and are most likely produced by the microbial oxidation of NH(4)(+) coming from clay minerals into NO(2)(-) and NO(3)(-). These metabolic pathways also produce NO. Subsequent dark abiotic reactions lead to NO(2) and HONO production. The snow cover is therefore not only an active photochemical reactor but also a biogeochemical reactor active in the cycling of nitrogen and it can affect atmospheric composition all year round. PMID- 20000752 TI - Effect of carbon concentration on changing the morphology of titanium carbide nanoparticles from cubic to cuboctahedron. AB - Titanium carbide nanoparticles were synthesized by flowing methane through a plasma generated from an arc discharge between two titanium electrodes. Different methane concentrations were employed in studies made to investigate the effects of carbon concentration on particle morphology. Transmission electron microscopy and X-ray diffraction were used to investigate the synthesized TiC nanopowders, whereupon it was found that nanocrystalline TiC nanoparticles prefer a cubic morphology at low concentrations of methane and a cuboctahedron morphology at high concentration of methane. The change in particle morphology is attributed to carbon affecting the relative growth rates of the {111} and {100} facets on a TiC seed crystal. PMID- 20000749 TI - New strategies for fluorescent probe design in medical diagnostic imaging. PMID- 20000753 TI - Ni(II)-catalyzed enantioselective conjugate addition of acetylenes to alpha,beta enones. AB - Alkynylaluminum reagents undergo enantioselective conjugate addition to cyclic alpha,beta-enones in the presence of chiral bisphosphine complexes of Ni(II). PMID- 20000754 TI - Steering organizational and conformational surface chirality by controlling molecular chemical functionality. AB - Molecular chirality on surfaces has been widely explored, both for intrinsically chiral molecules and for prochiral molecules that become chiral upon adsorption due to the reduced symmetry which follows from surface confinement. However, little attention has been devoted to chiral effects that originate from conformational degrees of freedom for adsorbed molecules. Here we have used scanning tunneling microscopy to investigate the self-assembled structures formed when a class of six linear, organic molecules (oligo-phenylene-ethynylenes) are adsorbed on a Au(111) surface under ultrahigh vacuum conditions. All of the investigated compounds are intrinsically achiral, but most display conformational chirality in the sense that the molecules can adsorb on the surface in different conformations giving rise to either one of two chiral surface enantiomers or a mirror-symmetric achiral meso form. A total of eleven observed adsorption structures are systematically investigated with respect to conformational chirality as well as point chirality (arising where molecular adsorption locally breaks the substrate symmetry) and organizational chirality (arising from the tiling pattern of the molecular backbones). A number of interesting correlations are identified between these different levels of chirality. Most importantly, we demonstrate that it is possible through control of the terminal group functionalization to steer the oligo(phenylene-ethynylene) molecular backbones into surface assemblies that either display pronounced organizational chirality or have mirror symmetric tiling patterns, and that it is furthermore possible to control the conformational surface chirality so the compounds preferentially assume either chiral or achiral surface conformers. PMID- 20000755 TI - Nanoscale heterostructures with molecular-scale single-crystal metal wires. AB - Creating nanoscale heterostructures with molecular-scale (<2 nm) metal wires is critical for many applications and remains a challenge. Here, we report the first time synthesis of nanoscale heterostructures with single-crystal molecular-scale Au nanowires attached to different nanostructure substrates. Our method involves the formation of Au nanoparticle seeds by the reduction of rocksalt AuCl nanocubes heterogeneously nucleated on the substrates and subsequent nanowire growth by oriented attachment of Au nanoparticles from the solution phase. Nanoscale heterostructures fabricated by such site-specific nucleation and growth are attractive for many applications including nanoelectronic device wiring, catalysis, and sensing. PMID- 20000756 TI - Tailored TiO2-SrTiO3 heterostructure nanotube arrays for improved photoelectrochemical performance. AB - TiO(2) nanotube arrays formed on Ti substrate by electrochemical anodization have been converted into TiO(2)-SrTiO(3) heterostructures by controlled substitution of Sr under hydrothermal conditions. The growth of SrTiO(3) crystallites on the nanotube array electrode was probed by electron microscopy and X-ray diffraction. As the degree of Sr substitution increases with the duration of hydrothermal treatment, an increase in the size of SrTiO(3) crystallites was observed. Consequently, with increasing SrTiO(3) fraction in the TiO(2)-SrTiO(3) nanotube arrays, we observed a shift in the flat band potential to more negative potentials, thus confirming the influence of SrTiO(3) in the modification of the photoelectrochemical properties. The TiO(2)-SrTiO(3) composite heterostructures obtained with 1 h or less hydrothermal treatment exhibit the best photoelectrochemical performance with nearly 100% increase in external quantum efficiency at 360 nm. The results presented here provide a convenient way to tailor the photoelectrochemical properties of TiO(2)-SrTiO(3) nanotube array electrodes and employ them for dye- or quantum-dot-sensitized solar cells and/or photocatalytic hydrogen production. PMID- 20000757 TI - Antibodies raised against chlamydial lipopolysaccharide antigens reveal convergence in germline gene usage and differential epitope recognition. AB - The structures of antigen-binding fragments from two related monoclonal antibodies have been determined to high resolution in the presence of several carbohydrate antigens raised against chlamydial lipopolysaccharide. With the exception of CDR H3, antibodies S54-10 and S73-2 are both derived from the same set of germline gene segments as the previously reported structures S25-2 and S45 18. Despite this similarity, the antibodies differ in specificity and the mechanism by which they recognize their cognate antigen. S54-10 uses an unrelated CDR H3 to recognize its antigen in a fashion analogous to S45-18; however, S73-2 recognizes the same antigen as S45-18 and S54-10 in a wholly unrelated manner. Together, these antibody-antigen structures provide snapshots into how the immune system uses the same set of inherited germline gene segments to generate multiple possible specificities that allow for differential recognition of epitopes and how unrelated CDR H3 sequences can result in convergent binding of clinically relevant bacterial antigens. PMID- 20000759 TI - Fabrication of disposable topographic silicon oxide from sawtoothed patterns: control of arrays of gold nanoparticles. AB - Disposable topographic silicon oxide patterns were fabricated from polymeric replicas of sawtoothed glass surfaces, spin-coating of poly(dimethylsiloxane) (PDMS) thin films, and thermal annealing at certain temperature and followed by oxygen plasma treatment of the thin PDMS layer. A simple imprinting process was used to fabricate the replicated PDMS and PS patterns from sawtoothed glass surfaces. Next, thin layers of PDMS films having different thicknesses were spin coated onto the sawtoothed PS surfaces and annealed at 60 degrees C to be drawn the PDMS into the valley of the sawtoothed PS surfaces, followed by oxygen plasma treatment to fabricate topographic silicon oxide patterns. By control of the thickness of PDMS layers, silicon oxide patterns having various line widths were fabricated. The silicon oxide topographic patterns were used to direct the self assembly of polystyrene-block-poly(2-vinylpyridine) (PS-b-P2VP) block copolymer thin films via solvent annealing process. A highly ordered PS-b-P2VP micellar structure was used to let gold precursor complex with P2VP chains, and followed by oxygen plasma treatment. When the PS-b-P2VP thin films containing gold salts were exposed to oxygen plasma environments, gold salts were reduced to pure gold nanoparticles without changing high degree of lateral order, while polymers were completely degraded. As the width of trough and crest in topographic patterns increases, the number of gold arrays and size of gold nanoparticles are tuned. In the final step, the silicon oxide topographic patterns were selectively removed by wet etching process without changing the arrays of gold nanoparticles. PMID- 20000758 TI - Structure and activity of lysozyme on binding to ZnO nanoparticles. AB - The interaction between ZnO nanoparticles (NPs) and lysozyme has been studied using calorimetric as well as spectrophotometric techniques, and interpreted in terms of the three-dimensional structure. The circular dichroism spectroscopic data show an increase in alpha-helical content on interaction with ZnO NPs. Glutaraldehyde cross-linking studies indicate that the monomeric form occurs to a greater extent than the dimer when lysozyme is conjugated with ZnO NPs. The enthalpy-driven binding between lysozyme and ZnO possibly involves the region encompassing the active site in the molecule, which is also the site for the dimer formation in a homologous structure. The enzyme retains high fraction of its native structure with negligible effect on its activity upon attachment to NPs. Compared to the free protein, lysozyme-ZnO conjugates are more stable in the presence of chaotropic agents (guanidine hydrochloride and urea) and also at elevated temperatures. The possible site of binding of NP to lysozyme has been proposed to explain these observations. The stability and the retention of a higher level of activity in the presence of the denaturing agent of the NP conjugated protein may find useful applications in biotechnology ranging from diagnostic to drug delivery. PMID- 20000760 TI - Enantioselective synthesis of 3,4-dihydroisoquinolin-1(2H)-ones by nickel catalyzed denitrogenative annulation of 1,2,3-benzotriazin-4(3H)-ones with allenes. AB - A denitrogenative annulation reaction of 1,2,3-benzotriazin-4(3H)-ones with allenes catalyzed by a nickel-phosphine complex to produce a variety of substituted 3,4-dihydroisoquinolin-1(2H)-ones in a regioselective manner is described. A highly enantioselective version, as well as structural evidence for the mechanistic course of this reaction, is also presented. PMID- 20000761 TI - Superoleophobic behavior induced by nanofeatures on oleophilic surfaces. AB - The control of surface wetting properties to produce robust and strong hydrophobic and oleophobic effects on intrinsically oleophilic surfaces is at the heart of many technological applications. In this paper, we explore the conditions to observe such effects when the roughness of the substrate is of fractal nature and consists of nanofeatures obtained by the ion track etching technique. The wetting properties were investigated using eight different liquids with surface tensions gamma varying from 18 to 72 mN m(-1). While it is observed that all the tested oils readily wet the flat substrates, it is found that the contact angles are systematically exalted on the rough surfaces even for the liquids with very low surface tension. For liquids with gamma > or = 25 mN m(-1) an oleophobic behavior is clearly induced by the nanostructuration. For liquids with gamma < 25 mN m(-1), although the contact angle is enhanced on the nanorough surfaces, it conserves its oleophilic character (theta* lower than 90 degrees). Moreover, our experiments show that even in the case of hexane, liquid having the lowest surface tension, the homogeneous wetting (Wenzel state) is never reached. This high resistance to liquid impregnation is discussed within the framework of recent approaches explaining the wetting properties of superoleophobic surfaces. PMID- 20000762 TI - The missing link between molecular triplets and spin-polarized free radicals: room temperature triplet states of nanocrystalline radical pairs. AB - Photochemical reactions of organic molecules in the solid state have an excellent potential in green chemistry technologies as they often proceed in high yields to give a single product without generating volatile organic solvent waste. While recent synthetic applications highlight the need for a better understanding of reaction mechanisms and kinetics, spectroscopic observations of excited states and short-lived intermediates in single crystals and powdered samples have been extremely challenging due to the high optical density and scattering power of single crystals and powdered samples. In this communication, we report the first direct observation of a radical pair triplet state by time-resolved electron paramagnetic resonance (TREPR) with nanocrystals of 4,4'-dimethoxy-dicumyl ketone (1OMe) suspended in water. Steady state irradiation of 1OMe had previously shown that reactions in dry powders and nanocrystalline suspensions proceed with high efficiency and chemoselectivity to generate 4,4'-dimethoxy-dicumene 2OMe by decarbonylation and radical coupling. The nanocrystalline suspensions were excited with an 25 ns laser pulse at 308 nm using a flow system within the cavity of a time-resolved EPR spectrometer. The resulting TREPR spectra showed strong spin polarization with enhanced absorption A and emission E signals in an AAAEEE pattern characteristic of a randomly oriented triplet with zero-field splitting parameters D = 243 G and E = 11 G as well as an isotropic exchange integral J = 45,000 G. The assignment of this spectrum to a radical pair triplet state was supported by measurements carried out in fluid solution and in frozen glasses, which allowed for the characterization of the free radical and the triplet excited molecular state of the starting ketone 1OMe. PMID- 20000763 TI - Tyrosine analogues for probing proton-coupled electron transfer processes in peptides and proteins. AB - A series of amino acids analogous to tyrosine, but differing in the physicochemical properties of the aryl alcohol side chain, have been prepared and characterized. These compounds are expected to be useful in understanding the relationships between structure, thermodynamics, and kinetics in long-range proton-coupled electron transfer processes in peptides and proteins. Systematic changes in the acidity, redox potential, and O-H bond strength of the tyrosine side chain could be induced upon substituting the phenol for pyridinol and pyrimidinol moieties. Further modulation was possible by introducing methyl and t butyl substitution in the position ortho to the phenolic hydroxyl. The unnatural amino acids were prepared by Pd-catalyzed cross-coupling of the corresponding halogenated aryl alcohol protected as their benzyl ethers with an organozinc reagent derived from N-Boc L-serine carboxymethyl ester. Subsequent debenzylation by catalytic hydrogenation yielded the tyrosine analogues in good yield. Spectrophotometric titrations revealed a decrease in tyrosine pK(a) of ca. 1.5 log units per included nitrogen atom, along with a corresponding increase in the oxidation (peak) potentials of ca. 200 mV, respectively. All told, the six novel amino acids described here have phenol-like side chains with pK(a)'s that span a range of 7.0 to greater than 10, and an oxidation (peak) potential range of greater than 600 mV at and around physiological pH. Radical equilibration EPR experiments were carried out to reveal that the O-H bond strengths increase systematically upon nitrogen incorporation (by ca. 0.5-1.0 kcal/mol), and radical stability and persistence increase systematically upon introduction of alkyl substitution in the ortho positions. The EPR spectra of the aryloxyl radicals derived from tyrosine and each of the analogues could be determined at room temperature, and each featured distinct spectral properties. The uniqueness of their spectra will be helpful in discerning one type of aryloxyl in the presence of other possible aryloxyl radicals in peptides and proteins with multiple tyrosine residues between which electrons and protons can be transferred. PMID- 20000764 TI - Diarylprolinol silyl ether salts as new, efficient, water-soluble, and recyclable organocatalysts for the asymmetric Michael addition on water. AB - A novel strategy for the catalytic asymmetric Michael addition of aldehydes to nitroolefins on water has been developed and provided the Michael adducts in excellent diastereo- and enantioselectivities. A notable feature of these organocatalysts is that they can be recycled for more than six times without a significant loss of catalytic activity and stereochemical control. In addition, the synthetic procedure presented is simple, practical, and environmentally benign. PMID- 20000765 TI - Zn2+-triggered amide tautomerization produces a highly Zn2+-selective, cell permeable, and ratiometric fluorescent sensor. AB - It is still a significant challenge to develop a Zn(2+)-selective fluorescent sensor with the ability to exclude the interference of some heavy and transition metal (HTM) ions such as Fe(2+), Co(2+), Ni(2+), Cu(2+), Cd(2+), and Hg(2+). Herein, we report a novel amide-containing receptor for Zn(2+), combined with a naphthalimide fluorophore, termed ZTRS. The fluorescence, absorption detection, NMR, and IR studies indicated that ZTRS bound Zn(2+) in an imidic acid tautomeric form of the amide/di-2-picolylamine receptor in aqueous solution, while most other HTM ions were bound to the sensor in an amide tautomeric form. Due to this differential binding mode, ZTRS showed excellent selectivity for Zn(2+) over most competitive HTM ions with an enhanced fluorescence (22-fold) as well as a red shift in emission from 483 to 514 nm. Interestingly, the ZTRS/Cd(2+) complex showed an enhanced (21-fold) blue-shift in emission from 483 to 446 nm. Therefore, ZTRS discriminated in vitro and in vivo Zn(2+) and Cd(2+) with green and blue fluorescence, respectively. Due to the stronger affinity, Zn(2+) could be ratiometrically detected in vitro and in vivo with a large emission wavelength shift from 446 to 514 nm via a Cd(2+) displacement approach. ZTRS was also successfully used to image intracellular Zn(2+) ions in the presence of iron ions. Finally, we applied ZTRS to detect zinc ions during the development of living zebrafish embryos. PMID- 20000766 TI - Electronic properties of pentacene versus triisopropylsilylethynyl-substituted pentacene: environment-dependent effects of the silyl substituent. AB - Energy measures of the intra- and intermolecular electronic effects of triisopropylsilylethynyl substitution on pentacene have been obtained from the combination of closely related gas phase and solid phase ultraviolet photoelectron spectroscopy (UPS) measurements along with solution electrochemical measurements. The results show that the shift to lower ionization energy that is expected with this substitution and observed in the gas phase measurements becomes negligible in solution and is even reversed in the solid phase. The principles that emerge from this analysis are supported by electronic structure calculations at the density functional theory level. The relation between the gas phase and solid phase UPS measurements illustrated here provides a general approach to investigating the electronic effects acting on molecules in the condensed phase, which in this case are greater than the direct substituent electronic effects within the molecule. Electronic properties such as lower ionization energies built into the single-molecule building blocks of materials and devices may be reversed in the solid state. PMID- 20000767 TI - Synthesis, structure, and bonding of stable complexes of pentavalent uranyl. AB - Stable complexes of pentavalent uranyl [UO(2)(salan-(t)Bu(2))(py)K](n) (3), [UO(2)(salan-(t)Bu(2))(py)K(18C6)] (4), and [UO(2)(salophen (t)Bu(2))(thf)]K(thf)(2)}(n) (8) have been synthesized from the reaction of the complex {[UO(2)py(5)][KI(2)py(2)]}(n) (1) with the bulky amine-phenolate ligand potassium salt K(2)(salan-(t)Bu(2)) or the Schiff base ligand potassium salt K(2)(salophen-(t)Bu(2)) in pyridine. They were characterized by NMR, IR, elemental analysis, single crystal X-ray diffraction, UV-vis spectroscopy, cyclic voltammetry, low-temperature EPR, and variable-temperature magnetic susceptibility. X-ray diffraction shows that 3 and 8 are polymeric and 4 is monomeric. Crystals of the monomeric complex [U(V)O(2)(salan (t)Bu(2))(py)][Cp*(2)Co], 6, were also isolated from the reduction of [U(VI)O(2)(salan-(t)Bu(2))(py)], 5, with Cp*(2)Co. Addition of crown ether to 1 afforded the highly soluble pyridine stable species [UO(2)py(5)]I.py (2). The measured redox potentials E(1/2) (U(VI)/U(V)) are significantly different for 2 ( 0.91 and -0.46 V) in comparison with 3, 4, 5, 7 and 9 (in the range -1.65 to 1.82 V). Temperature-dependent magnetic susceptibility data are reported for 4 and 7 and give mu(eff) of 2.20 and 2.23 mu(B) at 300 K respectively, which is compared with a mu(eff) of 2.6(1) mu(B) (300 K) for 2. Complexes 1 and 2 are EPR silent (4 K) while a rhombic EPR signal (g(x) = 1.98; g(y) = 1.25; g(z) = 0.74 (at 4 K) was measured for 4. The magnetic and the EPR data can be qualitatively analyzed with a simple crystal field model where the f electron has a nonbonding character. However, the temperature dependence of the magnetic susceptibility data suggests that one or more excited states are relatively low-lying. DFT studies show unambiguously the presence of a significant covalent contribution to the metal-ligand interaction in these complexes leading to a significant lowering of the pi(u)*. The presence of a back-bonding interaction is likely to play a role in the observed solution stability of the [UO(2)(salan-(t)Bu(2))(py)K] and [UO(2)(salophen-(t)Bu(2))(py)K] complexes with respect to disproportionation and hydrolysis. PMID- 20000768 TI - Laser diode thermal desorption/atmospheric pressure chemical ionization tandem mass spectrometry analysis of selected steroid hormones in wastewater: method optimization and application. AB - A rapid and reliable method enabling high-throughput sample analysis for quicker data generation, detection, and monitoring of eight selected steroid hormones in water matrixes was developed and validated. Our approach is based on a novel sample introduction method, the laser diode thermal desorption/atmospheric pressure chemical ionization (LDTD/APCI) coupled to tandem mass spectrometry (MS/MS). The optimization of instrumental parameters and a method application are presented. Our method was successfully applied to spiked effluent wastewater in the low-nanogram per liter concentrations with total analysis time reduced to seconds (15 s) using LDTD/APCI-MS/MS compared to minutes with traditional liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS) following solid phase extraction (SPE). The instrumental detection limits for LDTD/APCI-MS/MS ranged from 5 to 24 microg L(-1) and from 13 to 43 ng L(-1) for the method detection limits. Calibration curves in wastewater matrix showed good linearity (R(2) > 0.99), and precision (intraday and interday) was below 20%. This work demonstrates that LDTD/APCI-MS/MS could be used for fast and effective quantitative analysis of emerging contaminants in different water matrixes with reduced cost by eliminating the chromatography step used in traditional LC-MS/MS. PMID- 20000770 TI - Use of the hue parameter of the hue, saturation, value color space as a quantitative analytical parameter for bitonal optical sensors. AB - The hue or H component of the hue, saturation, value (HSV) color space has been studied as a quantitative analytical parameter for bitonal optical sensors. The robust nature of this parameter provides superior precision for the measurement of sensors which change colors with the speciation of some indicator molecule. This parameter has been compared to red, green, blue (RGB) intensity and RGB absorbance along with differences and ratios of both intensity and absorbance and has been demonstrated to be 2 to 3 times superior. The H value maintains this superior precision with variations in indicator concentration, membrane thickness, detector spectral responsivity, and illumination. Because this parameter is stable, simple to calculate, easily obtained from commercial devices such as scanners and digital cameras, continuous over the entire color gamut, and bound between values of 0 and 1, it shows great promise for use in a variety of sensing applications including imaging, automated analysis, pharmaceutical sensing, lab-on-a-chip devices, and quality control applications. PMID- 20000769 TI - Laser-induced acoustic desorption coupled with a linear quadrupole ion trap mass spectrometer. AB - In recent years, laser-induced acoustic desorption (LIAD) coupled with a Fourier transform ion cyclotron resonance (FT-ICR) mass spectrometer has been demonstrated to provide a valuable technique for the analysis of a wide variety of nonvolatile, thermally labile compounds, including analytes that could not previously be analyzed by mass spectrometry. Although FT-ICR instruments are very powerful, they are also large and expensive and, hence, mainly used as research instruments. In contrast, linear quadrupole ion trap (LQIT) mass spectrometers are common due to several qualities that make these instruments attractive for both academic and industrial settings, such as high sensitivity, large dynamic range, and experimental versatility. Further, the relatively small size of the instruments, comparatively low cost, and the lack of a magnetic field provide some distinct advantages over FT-ICR instruments. Hence, we have coupled the LIAD technique with a commercial LQIT, the Thermo Fischer Scientific LTQ mass spectrometer. The LQIT was modified for a LIAD probe by outfitting the removable back plate of the instrument with a 6 in. ConFlat flange (CFF) port, gate valve, and sample lock. Reagent ions were created using the LQIT's atmospheric pressure ionization source and trapped in the mass analyzer for up to 10 s to allow chemical ionization reactions with the neutral molecules desorbed via LIAD. These initial experiments focused on demonstrating the feasibility of performing LIAD in the LQIT. Hence, the results are compared to those obtained using an FT-ICR mass spectrometer. Despite the lower efficiency in the transfer of desorbed neutral molecules into the ion trap, and the smaller maximum number of available laser pulses, the intrinsically higher sensitivity of the LQIT resulted in a higher sensitivity relative to the FT-ICR. PMID- 20000771 TI - Single molecule imaging of protein molecules in nanopores. AB - The interactions between single protein molecules and nanoporous polycarbonate membranes were investigated at the single molecule level. Entrapment of proteins was shown to be size selective and was dependent on the membrane pore diameter. A pore size that is only slightly larger than the maximum dimension of the proteins was inadequate for intrusion into the pores. For a given protein, the number of molecules found at a given depth decreased as the pore size decreased. In addition, as the depth increased, for a given size pore, the number of molecules decreased rapidly. The depth-dependent histograms nicely fit a one-dimensional diffusion model. However, a highly restricted motion was observed even when the pore diameter was 10 times the size of the protein, resulting in anomalously small diffusion coefficients. We also demonstrated the subtle differences in depth distribution among BSA and hemoglobin that have nearly the same molecular weight but slightly different molecular shapes. These results give unique insights into the detailed mechanism of size-exclusion chromatography and membrane filtration. PMID- 20000772 TI - Fluorous synthesis of substituted sclerotigenin library. AB - A fluorous linker-assisted synthetic protocol has been developed for preparation of sclerotigenin-type benzodiazepine-quinazolinone library containing 144 analogues. Amide coupling of fluorous trimethoxybenzyl (TMB)-protected amino esters with anthranilic acids followed by base-promoted cyclizations afforded 4 benzodiazepine-2,5-diones. Further derivatization of benzodiazepinediones by reacting with azidobenzoyl chlorides, cyclization, and fluorous linker cleavage afforded the desired compound library. The reaction intermediates were purified by fluorous solid-phase extraction (F-SPE) and final products were further purified by prep-HPLC. PMID- 20000773 TI - Control of olive oil adulteration with copper-chlorophyll derivatives. AB - The present work proposes an analytical method able to detect in an adulterated olive oil sample the addition of the copper complexes of chlorophylls (E 141i). The method consists of a pigment extraction in liquid phase and subsequent analysis by HPLC-DAD. The profile of chlorophyll pigments of an olive oil is determined essentially by its content in pheophytins (a and b), but in no case any copper derivative. Different samples of colorant E 141i have been analyzed, the natural coloring additives used to adulterate vegetable oils. The 99.59+/ 0.52% of the chlorophyll pigments present in the different samples of E 141i colorant are not those of an olive oil (more than 75% are cupro-derivatives). Thus, the simple detection of one of the compounds in an olive oil indicates adulteration. The major chlorophyll derivative in all the E 141i colorants samples is Cu-pyropheophytin a and its limit of detection (LOD) defined at a signal-to-noise ratio of about 3 was 6.58 ng/g. PMID- 20000774 TI - Nematodetoxic aurovertin-type metabolites from a root-knot nematode parasitic fungus Pochonia chlamydosporia. AB - Chemical investigation of one fungal strain P. chlamydosporia YMF 1.00613 isolated from root knots of tobacco infected by Meloidogyne incognita led to the isolation and identification of four aurovertin-type metabolites, which include a new compound, aurovertin I (A1), and three known metabolites, aurovertins E, F and D (A2-A4). Their structures were established by spectroscopic studies such as 1D- and 2D-NMR and MS analysis. Aurovertin I (A1) is the first natural product with an aurovertin skeleton with one less carbon. Compounds A3 and A4 showed the toxicity to the worms of the free-living nematode Panagrellus redivevus with the LC(50) values 88.6 and 41.7 microg/mL at 48 h, respectively. All four aurovertins did not show obvious inhibitory effects on egg hatch of root knot nematode Meloidogyne incognita. The results suggested that the aurovertin-type metabolites produced by P. chlamydosporia might be one of the pathogenic factors involved in the suppression of nematodes. PMID- 20000775 TI - Design, multicomponent synthesis, and bioactivities of novel neonicotinoid analogues with 1,4-dihydropyridine scaffold. AB - Novel neonicotinoid analogues bearing a 1,4-dihydropridine scaffold were designed and synthesized by multicomponent reactions (MCRs) to enhance pi-pi stacking. The synthesized compounds were identified by (1)H NMR, (13)C NMR, high-resolution mass spectroscopy, and elemental analysis. Bioassay tests showed that some of them exhibited high insecticidal activities against pea aphid ( Aphis craccivora ). PMID- 20000776 TI - Design, synthesis, and antihepatitis B virus activities of novel 2-pyridone derivatives. AB - A series of novel 2-pyridone derivatives were synthesized and evaluated for their antihepatitis B virus (HBV) activity and cytotoxicity in vitro. Moderate to good activity against HBV DNA replication was observed in these 2-pyridone analogues. The most active compounds were 5d and 6l, with good inhibitory activity against HBV DNA replication (IC(50) = 0.206 and 0.12 microM, respectively) and remarkable high selectivity (selectivity indexes of >532 and 467, respectively). A pharmacophore model of the synthesized compounds was proposed by the GASP program. The pharmacophore model consists of three hydrophobic points, four HBA points, and one HBD point. The 2-pyridone derivatives represent a novel class of HBV inhibitors, which are worth further optimization. PMID- 20000777 TI - Cluster-bound nitriles do not click with organic azides: unexpected formation of imino complexes of the [Re(6)(mu(3)-Se)(8)](2+) core-containing clusters. AB - The reactions of C(6)H(5)CH(CH(3))N(3) with nitrile solvates of the [Re(6)(mu(3) Se)(8)](2+) core-containing cluster, [Re(6)(mu(3)-Se)(8)(PEt(3))(5)(MeCN)](2+)(1) and cis-[Re(6)(mu(3)-Se)(8)(PEt(3))(4)(MeCN)(2)](2+) (2), afforded the corresponding cationic imino complexes [Re(6)(mu(3)-Se)(8)(PEt(3))(5)(PhN horizontal lineCHCH(3))](2+) (3) and cis-[Re(6)(mu(3)-Se)(8)(PEt(3))(4)(PhN horizontal lineCHCH(3))(2)](2+) (4), respectively. Both compounds were spectroscopically and crystallographically characterized. A mechanism involving a 1,2-shift of one of the groups on the azido alpha-C atom of the cluster-azide intermediate concerted with the photoexpulsion of dinitrogen of the azido ligand is invoked to rationalize the formation of the imino complexes. Density functional theory calculations showed that a cluster-to-ligand transition was responsible for the absorption that promotes the photodecomposition of the cluster-azide complex. PMID- 20000778 TI - The selenazal drug ebselen potently inhibits indoleamine 2,3-dioxygenase by targeting enzyme cysteine residues. AB - The heme enzyme indoleamine 2,3-dioxygenase (IDO) plays an important immune regulatory role by catalyzing the oxidative degradation of l-tryptophan. Here we show that the selenezal drug ebselen is a potent IDO inhibitor. Exposure of human macrophages to ebselen inhibited IDO activity in a manner independent of changes in protein expression. Ebselen inhibited the activity of recombinant human IDO (rIDO) with an apparent inhibition constant of 94 +/- 17 nM. Optical and resonance Raman spectroscopy showed that ebselen altered the active site heme of rIDO by inducing a transition of the ferric heme iron from the predominantly high to low-spin form and by lowering the vibrational frequency of the Fe-CO stretch of the CO complex, indicating an opening of the distal heme pocket. Substrate binding studies showed that ebselen enhanced nonproductive l-tryptophan binding, while circular dichroism indicated that the drug reduced the helical content and protein stability of rIDO. Thiol labeling and mass spectrometry revealed that ebselen reacted with multiple cysteine residues of IDO. Removal of cysteine-bound ebselen with dithiothreitol reversed the effects of the drug on the heme environment and significantly restored enzyme activity. These findings indicate that ebselen inhibits IDO activity by reacting with the enzyme's cysteine residues that result in changes to protein conformation and active site heme, leading to an increase in the level of nonproductive substrate binding. This study highlights that modification of cysteine residues is a novel and effective means of inhibiting IDO activity. It also suggests that IDO is under redox control and that the enzyme represents a previously unrecognized in vivo target of ebselen. PMID- 20000779 TI - Global analysis of a plasmid-cured Shigella flexneri strain: new insights into the interaction between the chromosome and a virulence plasmid. AB - Shigella flexneri is an important human pathogen that causes dysentery, and remains a significant threat to public health, particularly in developing countries. The virulence of this pathogen is dependent on an acquired virulence plasmid. To investigate the crosstalk between the bacterial chromosome and the exogenous virulence plasmid, a virulence plasmid-cured strain was constructed using plasmid incompatibility. The global patterns of gene expression of this strain compared with the wild-type strain were analyzed using 2-DE combined with MALDI-TOF MS. Most known virulence factors of S. flexneri were identified in the 2-DE gels. Interestingly, the expression of the glycerol 3-phosphate (glp) regulon-encoded proteins was increased when the virulence plasmid was absent. Microarray analysis confirmed that regulation occurred at the transcriptional level. Purification and identification of DNA binding proteins with affinity for the regulatory region of the glp genes revealed that regulation mediated by the virulence plasmid to control the expression of the glp regulon might in turn be mediated by protein GlpR. To our knowledge, this is the first study analyzing the interaction between a pathogen chromosome and a virulence plasmid at the proteomic level. PMID- 20000780 TI - Dimeric guaianolides and sesquiterpenoids from Artemisia anomala. AB - A new dimeric guaianolide (8) and a new glaucolide (3), a seco-guaiaretic acid (4), two guaianolides (6 and 7), and five known sesquiterpene lactones (1, 2, 5, 9, and 10) were isolated from the aerial part of Artemisia anomala. Their structures were determined on the basis of chemical and spectroscopic analysis. In addition, their cytotoxic activities against five human cancer cell lines and anti-COX-2 effects in vitro were evaluated. PMID- 20000781 TI - Iridoids from Fraxinus excelsior with adipocyte differentiation-inhibitory and PPARalpha activation activity. AB - Two new secoiridoid glucosides, excelsides A (1) and B (2), were isolated from the seeds of Fraxinus excelsior. Their structures were elucidated as (2S,4S,3E) methyl 3-ethylidene-4-(2-methoxy-2-oxoethyl)-2-[(6-O-beta-D-glucopyranosyl-beta-d glucopyranosyl)oxy]-3,4-dihydro-2H-pyran-5-carboxylate and (2S,4S,3E)-methyl 3 ethylidene-4-{2-[2-(4-hydroxyphenyl)ethyl]oxy-2-oxoethyl}-2-[(6-O-beta-d glucopyranosyl-beta-d-glucopyranosyl)oxy]-3,4-dihydro-2H-pyran-5-carboxylate, respectively, on the basis of NMR and MS data. Eight known compounds were identified as nuzhenide (3), GI3 (4), GI5 (5), ligstroside (6), oleoside 11 methyl ester (7), oleoside dimethyl ester (8), 1'''-O-beta-D-glucosylformoside (9), and salidroside (10). Compounds 1-9 inhibited adipocyte differentiation in 3T3-L1 cells. Dilutions of the aqueous extract of F. excelsior (1:10,000) as well as compounds 2, 3, 4, 5, and 8 activated the peroxisome proliferator-mediated receptor-alpha (PPARalpha) reporter cell system in the range of 10(-4) M, compared to 10(-7)-10(-8) M for the synthetic PPARalpha activator, WY14,643. Both biological activity profiles support the hypothesis that inhibition of adipocyte differentiation and PPARalpha-mediated mechanisms might be relevant pathways for the antidiabetic activity of F. excelsior extract. PMID- 20000783 TI - Methylcobalamin's full- vs. "half"-strength cobalt-carbon sigma bonds and bond dissociation enthalpies: A >10(15) Co-CH3 homolysis rate enhancement following one-antibonding-electron reduction of methlycobalamin. AB - Methylcobalamin (MeCbl, MeB12) thermolyzed in ethylene glycol from 120 to 141 degrees C with 2,2,6,6-tetramethylpiperidinyl-1-oxy (TEMPO) as a Me(.) trap gives the homolysis products Co(II)B12 and TEMPO-Me quantitatively. The 5,6 dimethylbenzimidazole axial-base-off-base-on equilibrium in ethylene glycol has an enthalpy change of -5.1 (+/-2) kcal mol(-1) and an entropy change of -10.5 (+/ 4) cal mol(-1) K(-1), equilibrating between the 5,6-dimethylbenzimidazole coordinated base-on form and the two distinct yet similar non-coordinated forms: the base-off and the so-called "tuck-in" forms. The MeB12 Co-CH3 homolysis rates indicate an activation enthalpy of 41 +/- 3 kcal mol(-1), an activation entropy of 24 +/- 6 cal mol(-1) K(-1), and an estimated methylcob(III)alamin Co-CH3 bond dissociation energy of 37 +/- 3 kcal mol(-1). This is the strongest Co-C cobamide bond measured. Comparison of the MeCbl homolysis rate constant (10(-19 +/- 4) s( 1)) extrapolated to -30 degrees C with the known reduced-methylcobamide-radical anion values indicates rate enhancements of 10(22 +/- 4) (at -30 degrees C) following electrochemical reduction, or still over 10(15) at 25 degrees C. Such reduction provides an antibonding electron which weakens the Co-C bond from 37 kcal mol(-1) down to ca. 12 kcal mol(-1). These rate enhancements are greater than the analogous enzyme-induced Co-C cleavage rate enhancements in adenosylcobalamin (Coenzyme B12, AdoCbl)-dependent enzymes. However, electron transfer is not predicted for the mechanism of any adenosylcobalamin-dependent or methylcobalamin-dependent enzymes. PMID- 20000782 TI - A beta-carboline alkaloid from the Papua New Guinea marine sponge hyrtios reticulatus. AB - A new 1-imidazoyl-3-carboxy-6-hydroxy-beta-carboline alkaloid, named hyrtiocarboline (1), was isolated from a Papua New Guinea marine sponge, Hyrtios reticulatus. The structure was elucidated from spectroscopic data, including (1)H (15)N HMBC NMR experiments, which provided complementary (15)N chemical shift information in support of the structure. This compound showed selective antiproliferative activity against H522-T1 non-small cell lung, MDA-MB-435 melanoma, and U937 lymphoma cancer cell lines. PMID- 20000784 TI - Microbial oxidation of aromatics in enantiocontrolled synthesis. 2. Rational design of aza sugars (endo-nitrogenous). Total synthesis of +-kifunensine, mannojirimycin, and other glycosidase inhibitors. AB - A general method of synthesis for lactones and lactams related to carbohydrates has been developed that relies on the biocatalyticgeneration of 1-chloro-2,3 dihydroxycyclohexa-4,6-diene (l), obtained in excellent yield by fermentation of chlorobenzene with Pseudomonasputida 39D, followed by further functionalization to nitrogen-substituted cyclitols. These amino or azido cyclitols of type 15 are then subjected to controlled ozonolysis, which yields either lactones such as 27 or lactams containing five-membered (28) or six-membered (20 and 23) rings. Such compounds are useful intermediates for the preparation of aza sugars. Mannojirimycin (84 has been synthesized in seven steps from chlorobenzene. Kifunensine (7) has been prepared in 11 steps from chlorobenzene following an intersection with Hashimoto's procedure. Full experimental and spectral details are provided for all compounds. The potential of this general method and implications of the disclosed design features in the field of amino sugar and aza sugar synthesis are indicated. PMID- 20000785 TI - Chemically selective analysis of molecular monolayers by nonlinear optical stokes ellipsometry. AB - The application of nonlinear optical Stokes ellipsometry (NOSE) coupled with principal component analysis (PCA) is demonstrated for the chemically selective analysis of molecular monolayers. NOSE allows for rapid polarization measurements of nonlinear optical materials and thin surface films, which in turn benefits from comparably fast data analysis approaches. PCA combined with linear curve fitting techniques greatly reduce the analysis time relative to nonlinear curve fitting. NOSE-PCA is first validated with studies of z-cut quartz, followed by analysis of four thin dye films with similar nonlinear optical properties. The high precision of NOSE measurements combined with the rapid analysis time enabled chemical discrimination between different dyes and the practical realization of NOSE microscopy. PMID- 20000787 TI - Short and general procedure for synthesizing cis-1,2-fused 1,3-oxathiolan-, 1,3 oxaselenolan-, and 1,3-oxazolidin-2-imine carbohydrate derivatives. AB - Novel cis-1,2-fused 1,3-oxathiolan-, 1,3-oxaselenolan-, and 1,3-oxazolidin-2 imine carbohydrate derivatives have been prepared by treatment of the corresponding 1,2-anhydrosugars with potassium thiocyanate, potassium selenocyanate, and sodium cyanamide, respectively. The procedure is compatible with several protecting groups such as acyl, benzyl, and silyl and also with sugars of different configurations. PMID- 20000788 TI - Uncommon regioselectivity in the thiacalix[4]arene series: gross formylation of the cone conformer. AB - Thiacalix[4]arene immobilized in the cone conformation undergoes a direct Gross formylation reaction (Cl(2)CH-O-CH(3)/SnCl(4)/CH(2)Cl(2)) to give the upper-rim formylated thiacalixarene. Albeit using excess of the formylation agent and various reaction temperatures, only one formyl group is introduced into the meta position of the thiacalixarene skeleton. The surprising regioselectivity indicates dramatically different reactivity of the thiacalix[4]arene system when compared with a classical calix[4]arene analogue, which yields exclusively para isomers. The introduction of functional groups into the meta position represents an exceptional substitution pattern in thiacalixarene chemistry, which imparts an interesting conformational behavior to these compounds. The systematic NMR study revealed that the pinched cone-pinched cone equilibrium is remarkably shifted toward one pinched cone structure depending on the substitution. PMID- 20000790 TI - Nuclear magnetic resonance and infrared studies of N15-substituted trimethylsilyl , -germyl-, and -stannylanilines. A Test of (p->d)-pi interactions in group IV nitrogen bonds. AB - Infrared and proto n magnetic resonance spectra of the aniline-N14 and -N15 isotopomers and their N-trimethylsilyl, -germyl, and -stannyl derivatives have been measured and compared. Nmr assignments have been checked by comparisons of proton spectra at 60 and 100 Mcps; by proton-N15 heteronuclear experiments at 9400 G; and by observation of N15 satellites. The one-bond "5-H couplings can be interpreted (on the assumption of a dominant Fermi spin-spin interaction, small radial variations for nitrogen wave functions, and equal distributions of s character in the sigma bonds) in terms of pyramidal arrangements of bonds at nitrogen. The relation of the stereochemical situation at nitrogen to the question of (p-->d)-pi or (p-->p)-pi bonding involving nitrogen is discussed. PMID- 20000789 TI - Unprecedented rearrangement of 2-(2-aminoethyl)-1-aryl-3,4-dihydropyrazino[1,2 b]indazole-2-ium 6-oxides to 2,3-dihydro-1H-imidazo[1,2-b]indazoles. AB - Easily accessible 2-(2-aminoethyl)-1-aryl-3,4-dihydropyrazino[1,2-b]indazole-2 ium 6-oxides rearranged to 2,3-dihydro-1H-imidazo[1,2-b]indazoles under mild conditions. The rearrangement appeared to be general, tolerated a wide range of functional groups, and provided access to an as yet unexplored class of heterocycles. Herein we report the characterization of these heterocycles. PMID- 20000792 TI - gamma-Glutamyl PAMAM dendrimer as versatile precursor for dendrimer-based targeting devices. AB - Poly(amidoamine) (PAMAM) dendrimers are highly branched spherical polymers that have a unique surface of primary amine groups and provide a versatile design for targeted delivery of pharmaceuticals and imaging agents. Acetylation or succinylation of surface amine groups of PAMAM dendrimer derivatives is frequently performed to reduce nonspecific uptake. However, since targeting molecules, drugs/imaging agents, and acylating reagents react with the amine groups on dendrimer, such modification may limit the number of targeting molecules and/or drugs or may result in insufficient charge reduction. In this study, a gamma-glutamyl PAMAM dendrimer was designed and synthesized as a new precursor for targeting device. The relationship between surface electrical properties of the PAMAM dendrimer derivatives and pharmacokinetics was also determined. A PAMAM dendrimer (generation 4.0) was modified with a small number of Bolton-Hunter reagent to prepare Phe-P (pI 9.2). The amine residues of Phe-P were gamma-glutamylated to prepare Glu-P (pI 7.1). The alpha-amine residues of Glu-P were then acetylated or succinylated to prepare Ac-Glu-P (pI 5.3) or Suc Glu-P (pI 3.6). For comparison, Phe-P was acetylated or succinylated to prepare Ac-P (pI 6.0) or Suc-P (pI 5.1). All the PAMAM dendrimer derivatives exhibited similar molecular size (7.2 to 7.8 nm) except for Ac-P (5.1 nm). The biodistribution studies were performed after radioiodination of each PAMAM dendrimer derivative with Na[(125)I]I. When injected intravenously to mice, both [(125)I]Ac-P and [(125)I]Suc-P exhibited prolonged radioactivity levels in the blood and significantly lower hepatic and renal radioactivity levels than those of [(125)I]Phe-P. Both [(125)I]Glu-P and [(125)I]Ac-Glu-P showed residence times in the blood similar to those of [(125)I]Ac-P and [(125)I]Suc-P. However, [(125)I]Glu-P also registered higher radioactivity levels in the kidney. High hepatic and renal radioactivity levels were observed with highly anionic [(125)I]Suc-Glu-P. These results indicate that, while the manipulation of pI between 5 to 6 would be appropriate to enhance blood retention and reduce renal and hepatic uptake, the amount of primary amine residues on dendrimer surface may also play a crucial role in their renal uptake. The findings in this study show that gamma-glutamyl PAMAM dendrimers would constitute versatile precursors to prepare PAMAM dendrimer-based targeting devices due to their neutral molecular charge (pI 7.1) and the presence of a large number of alpha-amine residues available for conjugation of targeting molecules and drugs/imaging agents. PMID- 20000793 TI - Meldrum's acids and 5-alkylidene Meldrum's acids in catalytic carbon-carbon bond forming processes. AB - Meldrum's acid (2,2-dimethyl-1,3-dioxane-4,6-dione) is a molecule with a unique history, owing to its originally misassigned structure, as well as a unique place among acylating agents, owing to its high acidity and remarkable electrophilicity. In this Account, we outline the work of our group and others toward harnessing the reactivity of Meldrum's acid derivatives in catalytic C-C bond-forming reactions. Taking advantage of the ability of Meldrum's acid to decompose to CO(2) and acetone following acyl substitution, we have shown that intramolecular Friedel-Crafts acylations can be performed under mild Lewis acidic conditions to yield a variety of benzocyclic ketones. In a further expansion of this method, a domino Friedel-Crafts acylation/alpha-tert alkylation reaction was used to complete the first total synthesis of (+/-)-taiwaniaquinol B. The unique characteristics of Meldrum's acid extend to its alkylidene derivatives, which have also proven exceptionally useful for the development of new reactions not readily accessible from other unsaturated carbonyl electrophiles. By combining the electrophilicity and dienophilicity of alkylidene Meldrum's acid with our Friedel-Crafts chemistry, we have demonstrated new domino syntheses of coumarin derivatives and tetrahydrofluorenones by conjugate additions, Diels-Alder cycloadditions, and C-H functionalizations. Additionally, we have used these powerful acceptors to allow conjugate alkenylation with functionalized organostannanes, and conjugate allylation under very mild conditions. We have also shown that these molecules permit the asymmetric formation of all-carbon quaternary stereocenters via enantioselective conjugate additions. These reactions employ dialkylzinc nucleophiles, maximizing functional group compatibility, while the presence of a Meldrum's acid moiety in the product allows a variety of postaddition modifications. A full investigation of this reaction has determined the structural factors of the alkylidene that contribute to optimal enantioselectivity. We have also used these acceptors to form tertiary propargylic stereocenters in very high enantiomeric excess by an extremely mild, Rh(I)-catalyzed addition of TMS-acetylene. Overall, we demonstrate that Meldrum's acid and its derivatives provide access to a broad range of reactivities that, combined with their ease of handling and preparation, make them ideal electrophiles. PMID- 20000791 TI - Stabilization of the nitric oxide (NO) prodrugs and anticancer leads, PABA/NO and Double JS-K, through incorporation into PEG-protected nanoparticles. AB - We report the stabilization of the nitric oxide (NO) prodrugs and anticancer lead compounds, PABA/NO (O(2)-{2,4-dinitro-5-[4-(N-methylamino)benzoyloxy]phenyl} 1 (N,N-dimethylamino)diazen-1-ium-1,2-diolate) and "Double JS-K" 1,5-bis-{1-[(4 ethoxycarbonyl)piperazin-1-yl]diazen-1-ium-1,2-diol-2-ato}-2,4-dinitrobenzene, through their incorporation into polymer-protected nanoparticles. The prodrugs were formulated in block copolymer-stabilized nanoparticles with sizes from 220 to 450 nm by a novel rapid precipitation process. The block copolymers, with polyethylene glycol (PEG) soluble blocks, provide a steric barrier against NO prodrug activation by glutathione. Too rapid activation and NO release has been a major barrier to effective administration of this class of compounds. The nanoparticle stabilized PABA/NO are protected from attack by glutathione as evidenced by a significant increase in time taken for 50% decomposition from 15 min (unformulated) to 5 h (formulated); in the case of Double JS-K, the 50% decomposition time was extended from 4.5 min (unformulated) to 40 min (formulated). The more hydrophobic PABA/NO produced more stable nanoparticles and correspondingly more extended release times in comparison with Double JS-K. The hydrophobic blocks of the polymer were either polystyrene or polylactide. Both blocks produced nanoparticles of approximately the same size and release kinetics. This combination of PEG-protected nanoparticles with sizes appropriate for cancer targeting by enhanced permeation and retention (EPR) and delayed release of NO may afford enhanced therapeutic benefit. PMID- 20000794 TI - Nonbiofouling polymer brush with latent aldehyde functionality as a template for protein micropatterning. AB - A novel, nonfouling polymer brush, poly-N-[(2,3-dihydroxypropyl)acrylamide] (PDHPA), containing latent aldehyde groups, was synthesized by surface initiated atom transfer radical polymerization (SI-ATRP). The synthetic parameters were adjusted to produce brushes with varying graft densities and molecular weights. High-density PDHPA brushes successfully prevented the nonspecific protein adsorption from single protein solutions as well as from human platelet poor plasma. Patterns of nonfouling PDHPA and reactive PDHPA-aldehyde domains on the brush surface were created by a combination of photo and wet chemical lithography from a single homogeneous PDHPA brush. Successful micropatterning of single proteins and multiple proteins were achieved using this novel substrate. The high density brush prevented the diffusion of large proteins into the brush, while a monolayer of covalently coupled proteins was formed on the PDHPA-aldehyde domains. Atomic force microscopy (AFM) force measurements using a biotin coupled AFM tip showed that covalently coupled streptavidin retained its activity, while PDHPA domains showed little nonspecific adsorption of streptavidin. The current study avoids tedious and complicated synthetic processes employed in conventional approaches by providing a novel approach to protein micropatterning from a single, multifunctional polymer brush. PMID- 20000795 TI - Facile fabrication of uniform silica films with tunable physical properties using silicatein protein from sponges. AB - We report an elegant and simple method to fabricate uniform silica films with controlled thickness, roughness, and hydrophilicity using nanogram quantities of silicatein, a protein involved in silica synthesis in sponges. The formation of uniform silica films was achieved by immobilization of silicatein on gold-coated surfaces that had been functionalized with amines. Specifically, the amines, cystamine and cysteamine having disulfide and thiol groups, respectively, were bound to surfaces and treated with a cross-linking agent, glutardialdehyde (GDA) before protein immobilization. Silica was formed on the silicatein bound surfaces under environmentally benign conditions using tetramethoxysilane (TMOS). The thickness (20-100 nm), roughness (1.2-5.2 nm), and water contact angle (48 degrees-16 degrees) of the silica films could be controlled by varying the amount of silicatein adsorbed (10-30 ng/cm(2)) and time of exposure of protein-coated surfaces (30-120 min) to silica precursors. The silicatein protein retained around 90% of its intrinsic activity when attached to the functionalized surfaces with similar activity being observed for silica films formed from TMOS or tetraethoxysilane (TEOS). This simple route to prepare silica films of controlled physical properties could have potential application in membrane fabrication, biomedical devices, biosensors, and next generation electronic components. PMID- 20000796 TI - pH-controlled assembly and properties of LbL membranes from branched conjugated poly(alkoxythiophene sulfonate) and various polycations. AB - We report on multilayer layer-by-layer (LbL) films of the conjugated polymer sodium poly[2-(3-thienyl)ethyloxy-4-butylsulfonate] (PTH) assembled with polycations: poly(diallyldimethylammonium chloride) (PDDA), 20% quaternized poly(N-ethyl-4-vinylpyridinium bromide) (Q20), poly(ethylene imine) (PEI), and poly(allylamine hydrochloride) (PAH). These films were prepared through spin assisted LbL assembly under various pH conditions. We demonstrated a crucial role of the deposition pH in formation of PTH/polycation films and showed that decrease in the deposition pH from 7.5 to 2.5 limits the PTH multilayer formation to Q20/PTH and PDDA/PTH films due to reduced charge density in the poly(thiophene) chains. We show that optical and surface properties of the resulting PTH/polycation films can be tuned by varying a polycation component and/or by varying the deposition pH. The fluorescence properties of the Q20/PTH, PEI/PTH, and PDDA/PTH films are pH-dependent, and the films exhibit the drastic changes in photoluminescent intensity when transferred into solutions with different pH values, which may find useful in optical sensing applications. PMID- 20000797 TI - Statistical mechanics and dynamics of two supported stacked lipid bilayers. AB - The statistical physics and dynamics of double supported bilayers are studied theoretically. The main goal in designing double supported lipid bilayers is to obtain model systems of biomembranes: the upper bilayer is meant to be almost freely floating, the substrate being screened by the lower bilayer. The fluctuation-induced repulsion between membranes and between the lower membrane and the wall are explicitly taken into account using a Gaussian variational approach. It is shown that the variational parameters, the "effective" adsorption strength, and the average distance to the substrate, depend strongly on temperature and membrane elastic moduli, the bending rigidity, and the microscopic surface tension, which is a signature of the crucial role played by membrane fluctuations. The range of stability of these supported membranes is studied, showing a complex dependence on bare adsorption strengths. In particular, the experimental conditions of having an upper membrane slightly perturbed by the lower one and still bound to the surface are found. Included in the theoretical calculation of the damping rates associated with membrane normal modes are hydrodynamic friction by the wall and hydrodynamic interactions between both membranes. PMID- 20000798 TI - High aspect ratio constructive nanolithography with a photo-dimerizable molecule. AB - A major challenge in constructive nanolithography is the preservation of the lateral resolution of a monolayer-thick template pattern while amplifying it to a structure with a thickness above 10 nm. So far, the most successful approach to achieve this is surface-initiated polymerization (SIP) from e-beam structured monolayer templates in a multistep process. However, spreading of the polymer on the substrate leads to a rapid line-widening. Therefore, structures with lateral resolutions well below 100 nm and thicknesses above 10 nm (aspect ratio: 0.1) were not reported yet. Our approach of photoinduced, constructive, reversible nanolithography, is based on nanografting within a coumarin-derivative thiol (CDT) solution using the tip of an atomic force microscope (AFM). By photodimerization and the formation of disulfide bonds, the CDT polymerizes in a single-step process. We demonstrate the highest lateral resolution in constructive nanolithography at thicknesses above 10 nm (40 nm lateral resolution at 12 nm thickness, aspect ratio: 0.3). PMID- 20000799 TI - Self-catalytic sol-gel synergetic replication of uniform silica nanotubes using an amino acid amphiphile dynamically growing fibers as template. AB - The aggregation behavior of a novel histidine derivative surfactant N-dodecanoyl L-histidine (DHis) in aqueous solution was investigated. By mixing with cationic/anionic surfactant, an interesting phase transition over time from transparent homogeneous solution to jelly and finally to fibrous floccules was observed with aggregates growth taken place in the systems. Making use of the growing fibrous structures in the DHis/DTEAB and DHis/SDS systems as surface charge tunable dynamic template, we achieved sol-gel replication of silica and titania nanotubes in mild conditions, respectively. The possible mechanism underlying the formation of silica nanotubes was proved to be synergistically self-catalyzed hydrolysis and condensation of silica precursors depositing on the template surface. In particular, by optimizing the sol-gel process, silica nanotubes with tunable sizes in diameter and uniform wall thickness could be obtained in high yield. The large specific surface area and peculiar photoluminescence property of the silica nanomaterials indicated that the synthetic nanotubes were well-defined in both morphology and physical properties. Importantly, this fibrous aggregates could be universally used in opposite charged inorganic precursors to accomplish inorganic oxide nanotubes via sol-gel replication process, which may provide a general route for the construction of one-dimensional nanostructures in aqueous media. PMID- 20000800 TI - Decomposition of ethanol on Pd(111): a density functional theory study. AB - Ethanol decomposition over Pd(111) has been systematically investigated using self-consistent periodic density functional theory, and the decomposition network has been mapped out. The most stable adsorption of the involved species tends to follow the gas-phase bond order rules, wherein C is tetravalent and O is divalent with the missing H atoms replaced by metal atoms. Desorption is preferable for adsorbed ethanol, methane, and CO, while for the other species decomposition is preferred. For intermediates going along the decomposition pathways, energy barriers for the C-C, C(alpha)-H, and O-H scissions are decreased, while it is increased for the C-O path or changes less for the C(beta)-H path. For each of the C-C, C-O, and C-H paths, the Bronsted-Evans-Polanyi relation holds roughly. The most likely decomposition path is CH(3)CH(2)OH --> CH(3)CHOH --> CH(3)CHO --> CH(3)CO --> CH(2)CO --> CHCO --> CH + CO --> CO + H + CH(4) + C. PMID- 20000801 TI - Doping-induced conductivity transitions in molecular layers of polyaniline: optical studies of electronic state changes. AB - The doping-induced conductivity transitions in molecular layers of polyaniline have been studied by monitoring the correlated optical and spectroscopic changes using spectroscopic and single wavelength extinction ellipsometry, also in total internal reflection mode (TIRE), together with reflection spectrometry. The measurements were performed on deposited multilayers as well as on a Langmuir monolayer at the air-water interface, as a function of acidic doping. We found that the characteristic spectroscopic features of conducting and insulating polyaniline persisted down to the single layer, both in the solid state and at the air-water interface. We also investigated in real time the modulation of conductivity induced by the intercalation of Li ions in the polyaniline film, by a combination of time-resolved ellipsometry and reflectivity spectra measurements. In this case, the enhanced sensitivity provided by the TIRE geometry, combined with the relatively fast time scale accessible by the single wavelength ellipsometry, allowed us to follow in detail in real time the doping/dedoping process. PMID- 20000802 TI - Detection of oxidative damages on viral capsid protein for evaluating structural integrity and infectivity of human norovirus. AB - The infectivity evaluation of noncultivatable viruses, such as human norovirus, is crucial to address needs for ensuring the safety in usage of water and marine products. In this work, we tested a new approach to evaluate viral particle integrity, in which oxidatively produced carbonyl groups on viral capsid protein were quantitatively detected. As a result, the decrease in the infectivity of human astrovirus, a representative enteric virus, positively correlated with the amount of oxidative damage on viral particles. Furthermore, when human norovirus was treated by 1 ppm free chlorine for 15 min, 49.93% of virions were recovered as oxidatively damaged particles, which represents a 5-fold increase over those treated by 0.5 ppm free chlorine for 15 min. The detection of the carbonylated viral particles could be a powerful tool for the evaluation of the decrease in the infectivity of noncultivatable viruses. PMID- 20000803 TI - Mechanical control of ATP synthase function: activation energy difference between tight and loose binding sites. AB - Despite exhaustive chemical and crystal structure studies, the mechanistic details of how F(o)F(1)-ATP synthase can convert mechanical energy to chemical, producing ATP, are still not fully understood. On the basis of quantum mechanical calculations using a recent high-resolution X-ray structure, we conclude that formation of the P-O bond may be achieved through a transition state (TS) with a planar PO(3)(-) ion. Surprisingly, there is a more than 40 kJ/mol difference between barrier heights of the loose and tight binding sites of the enzyme. This indicates that even a relatively small change in active site conformation, induced by the gamma-subunit rotation, may effectively block the back reaction in beta(TP) and, thus, promote ATP. PMID- 20000804 TI - Evaluation and optimization of a force field for crystalline forms of mannitol and sorbitol. AB - Two force fields, the GROMOS53A5/53A6 (united atom) and the AMBER95 (all atom) parameter sets, coupled with partial atomic charges derived from quantum mechanical calculations were evaluated for their ability to reproduce the known crystalline forms of the polyols mannitol and sorbitol. The force fields were evaluated using molecular dynamics simulations at 10 K (which is akin to potential energy minimization) with the simulation cell lengths and angles free to evolve. Both force fields performed relatively poorly, not being able to simultaneously reproduce all of the crystal structures within a 5% deviation level. The parameter sets were then systematically optimized using sensitivity analysis, and a revised AMBER95 set was found to reproduce the crystal structures with less than 5% deviation from experiment. The stability of the various crystalline forms for each of the parameter sets (original and revised) was then assessed in extended MD simulations at 298 K and 1 bar covering 1 ns simulation time. The AMBER95 parameter sets (original and revised) were found to be effective in reproducing the crystal structures in these more stringent tests. Remarkably, the performance of the original AMBER95 parameter set was found to be slightly better than that of the revised set in these simulations at 298 K. The results of this study suggest that, whenever feasible, one should include molecular simulations at elevated temperatures when optimizing parameters. PMID- 20000805 TI - Planar plasmonic focusing and optical transport using CdS nanoribbon. AB - Planar plasmonic focusing of surface plasmon polaritons (SPPs) by an in-plane nanostructure consisting of Ag-column arrays and an in-plane Fresnel zone plate (FZP) with a Cu grating underneath for energy compensation was demonstrated. The CdS-based hybrid plasmonic waveguide generated in the Ag-column arrays was characterized with a scanning near-field optical microscope. By using the FZP focusing structure, the SPP modes were separated from the CdS photoluminescence background and focused at the FZP focus area, and in this way, were used as the source for the SPP waveguide. Finite-difference time-domain simulations correspond with the experimental observations, suggesting that this is indeed an effective approach to control SPP coupling within the dielectric nanoribbon waveguide. PMID- 20000806 TI - Indium triflate-assisted nucleophilic aromatic substitution reactions of nitrosobezene-derived cycloadducts with alcohols. AB - Exclusive nucleophilic aromatic substitution of hydrogen in conjunction with cleavage of the N-O bond was observed when nitrosobenzene-derived nitroso cycloadducts were treated with indium triflate in the presence of alcohols. Aromatic alkoxylated syn-1,4 aminocycloalkenol products with good to excellent regioselectivity (16:84 to 0:100, ortho:para) were obtained. PMID- 20000808 TI - Oligo- and polythiophene/ZnO hybrid nanowire solar cells. AB - We demonstrate the basic operation of an organic/inorganic hybrid single nanowire solar cell. End-functionalized oligo- and polythiophenes were grafted onto ZnO nanowires to produce p-n heterojunction nanowires. The hybrid nanostructures were characterized via absorption and electron microscopy to determine the optoelectronic properties and to probe the morphology at the organic/inorganic interface. Individual nanowire solar cell devices exhibited well-resolved characteristics with efficiencies as high as 0.036%, J(sc) = 0.32 mA/cm(2), V(oc) = 0.4 V, and a FF = 0.28 under AM 1.5 illumination with 100 mW/cm(2) light intensity. These individual test structures will enable detailed analysis to be carried out in areas that have been difficult to study in bulk heterojunction devices. PMID- 20000807 TI - Na(+) and K(+) allosterically regulate cooperative DNA binding by the human progesterone receptor. AB - Cooperativity is a common mechanism used by transcription factors to generate highly responsive yet stable gene regulation. For the two isoforms of human progesterone receptor (PR-A and PR-B), differences in cooperative DNA binding energetics may account for their differing transcriptional activation properties. Here we report on the molecular origins responsible for cooperativity, finding that it can be activated or repressed with Na(+) and K(+), respectively. We demonstrate that PR self-association and DNA-dependent cooperativity are linked to a monovalent cation binding event and that this binding is coupled to modulation of receptor structure. K(+) and Na(+) are therefore allosteric effectors of PR function. Noting that the apparent binding affinities of Na(+) and K(+) are comparable to their intracellular concentrations and that PR isoforms directly regulate the genes of a number of ion pumps and channels, these results suggest that Na(+) and K(+) may additionally function as physiological regulators of PR action. PMID- 20000810 TI - 1,214-Triazoles. XXII. Derivatives of the s-triazolo [3,4-alpha] phthalazine and related ring systems. PMID- 20000809 TI - Structure, mechanism, and substrate profile for Sco3058: the closest bacterial homologue to human renal dipeptidase . AB - Human renal dipeptidase, an enzyme associated with glutathione metabolism and the hydrolysis of beta-lactams, is similar in sequence to a cluster of approximately 400 microbial proteins currently annotated as nonspecific dipeptidases within the amidohydrolase superfamily. The closest homologue to the human renal dipeptidase from a fully sequenced microbe is Sco3058 from Streptomyces coelicolor. Dipeptide substrates of Sco3058 were identified by screening a comprehensive series of l Xaa-l-Xaa, l-Xaa-d-Xaa, and d-Xaa-l-Xaa dipeptide libraries. The substrate specificity profile shows that Sco3058 hydrolyzes a broad range of dipeptides with a marked preference for an l-amino acid at the N-terminus and a d-amino acid at the C-terminus. The best substrate identified was l-Arg-d-Asp (k(cat)/K(m) = 7.6 x 10(5) M(-1) s(-1)). The three-dimensional structure of Sco3058 was determined in the absence and presence of the inhibitors citrate and a phosphinate mimic of l-Ala-d-Asp. The enzyme folds as a (beta/alpha)(8) barrel, and two zinc ions are bound in the active site. Site-directed mutagenesis was used to probe the importance of specific residues that have direct interactions with the substrate analogues in the active site (Asp-22, His-150, Arg-223, and Asp-320). The solvent viscosity and kinetic effects of D(2)O indicate that substrate binding is relatively sticky and that proton transfers do not occurr during the rate-limiting step. A bell-shaped pH-rate profile for k(cat) and k(cat)/K(m) indicated that one group needs to be deprotonated and a second group must be protonated for optimal turnover. Computational docking of high-energy intermediate forms of l/d-Ala-l/d-Ala to the three-dimensional structure of Sco3058 identified the structural determinants for the stereochemical preferences for substrate binding and turnover. PMID- 20000812 TI - On the alkaloids of Strychnos. XXXV. The occurrence of akagerine in South American Strychnos. AB - Akagerine (3) was isolated from three different species of South American Strychnos: from S. gardneri A. D.C. (Rio de Janeiro, Brazil), together with 11 methoxydiaboline (2); from S. jobertiana Baillon (Brazil), together with diaboline (1): and from S. parvifolia D.C. (Bahia, Brazil). PMID- 20000814 TI - Field evaluation of polyurethane foam passive air samplers to assess airborne PAHs in occupational environments. AB - There is a need for simple air sampling techniques to enable routine monitoring of the occupational exposure to polycyclic aromatic hydrocarbons (PAHs) in compliance with occupational exposure limits. Other gas-phase contaminants can be monitored in workplaces using passive samplers but this is currently not the case for PAHs. Here, polyurethane foam (PUF) disk passive air samplers (PAS), routinely used for outdoor air monitoring of PAHs and POPs, were assessed for their suitability in an indoor occupational environment against: ability to accumulate detectable levels within 1-2 weeks; quantitative sampling of benzo(a)pyrene (BaP), precision, uptake kinetics, influence of shelter design, and performance of 16 deuterated PAHs as depuration compounds (DCs). Sampling rates (R-values) for PAHs in PUF-PAS, estimated by comparison to low-volume active samplers, and the loss of DCs, varied for individual PAHs (1-10 m(3) day( 1)) but were found to be in the same order of magnitude for both gas-phase and particle-associated PAHs including BaP. Only one PAH (Acy) fulfilled the DC criteria of >40% loss during the 2 week exposure. These results suggest that PUF PAS are potentially useful tools for PAHs in occupational environments in screening workplaces and identifying sources/hotspots - although unlikely to replace active sampling. PMID- 20000815 TI - Atmospheric stability of levoglucosan: a detailed laboratory and modeling study. AB - Levoglucosan, an important molecular marker for biomass burning, represents an important fraction of the water-soluble organic carbon in atmospheric particles influenced by residential wood burning and wildfires. However, particle phase oxidation processes of levoglucosan by free radicals are not well-known. Hence, detailed kinetic studies on the reactivity of levoglucosan with OH, NO(3), and SO(4)(-) radicals in aqueous solutions were performed to better understand the levoglucosan oxidation in the deliquescent particles. The data obtained were implemented into a parcel model with detailed microphysics and complex multiphase chemistry to investigate the degradation fluxes of levoglucosan in cloud droplets and in deliquescent particles. The model calculations show that levoglucosan can be oxidized readily by OH radicals during daytime with mean degradation fluxes of about 7.2 ng m(-3) h(-1) in summer and 4.7 ng m(-3) h(-1) in winter for a polluted continental plume. This indicates that the oxidation of levoglucosan in atmospheric deliquescent particles is at least as fast as that of other atmospherically relevant organic compounds and levoglucosan may not be as stable as previously thought in the atmosphere, especially under high relative humidity conditions. PMID- 20000816 TI - Identification and screening analysis of halogenated norbornene flame retardants in the Laurentian Great Lakes: Dechloranes 602, 603, and 604. AB - Dechlorane (Dec) 602, Dechlorane (Dec) 603, Dechlorane (Dec) 604, and Dechlorane Plus (DP) are flame retardant substitutes for mirex. Dec 602, 603, and 604 were detected in sediment and fish from the Laurentian Great Lakes. Lake Ontario surface sediments had the highest concentrations of Dec 602 and 604 at 6.0 and 4.0 ng/g dry weight, respectively. Temporal analysis of a Lake Ontario sediment core indicates that Dec 602 and 604 trends are similar to DP peaking in the early 1980s. Lake trout and whitefish from Lake Ontario also had the highest concentrations of Dec 602 and 604 at 34 and 1.2 ng/g lipid. Concentrations of Dec 602 were higher than those of DP in all fish samples, indicating that Dec 602 is likely more bioavailable and/or more readily bioaccumulates than DP. Spatial trends for Dec 602 and 604 in sediment and fish indicate that manufacturing plants along the Niagara River upstream of Lake Ontario were important sources of Dec 602 and 604 to the Great Lakes, while Dec 603 in the Great Lakes is likely from atmospheric deposition. The findings of this first report of Dec 602, 603, and 604 in the Laurentian Great Lakes basin suggests further investigation of halogenated norbornene flame retardants in the environment is merited. PMID- 20000817 TI - Growth of stacking-faults-free zinc blende GaAs nanowires on Si substrate by using AlGaAs/GaAs buffer layers. AB - Vertical GaAs nanowires on Si (111) substrate were grown by metal organic chemical vapor deposition via Au-catalyst vapor-liquid-solid mechanism. Stacking faults-free zinc blende nanowires were realized by using AlGaAs/GaAs buffer layers and growing under the optimized conditions, that the alloy droplet act as a catalyst rather than an adatom collector and its size and composition would keep stable during growth. The stable droplet contributes to the growth of stacking-faults-free nanowires. Moreover, by using the buffer layers, epitaxial growth of well-aligned NWs was not limited by the misfit strain induced critical diameter, and the unintentional doping of the GaAs nanowires with Si was reduced. PMID- 20000818 TI - Polybrominated diphenyl ethers in umbilical cord blood and relevant factors in neonates from Guiyu, China. AB - We aimed to evaluate the exposure of neonates to polybrominated diphenyl ethers (PBDEs) from a primitive e-waste (obsolete electrical and electronic devices) recycling area, in Guiyu, China, and a control area, Chaonan, China, through umbilical cord blood (UCB), the health effects, and relevant factors. Questionnaires were addressed, and UCB was collected shortly after birth from 153 pregnant women between May and July 2007. Blood samples were prepared by liquid liquid extracting methods. PBDE concentration was determined by gas chromatography/mass spectrometry in the electron capture negative ionization mode. The total PBDE concentration was higher in UCB samples from Guiyu than in Chaonan samples (median 13.84, range 1.14-504.97 ng g(-1) lipid, vs 5.23, range 0.29-363.70 ng g(-1) lipid) (p < 0.05). BDE-209 was the dominant PBDE congener, followed by BDE-47, -153, and -99. Residence in Guiyu, which is a site for e waste recycling, involvement in e-waste recycling, and the residence also being used as a family workshop were significant factors contributing to PBDE exposure. PBDE levels significantly differed in neonates by normal birth and adverse birth outcomes including stillbirth, low birth weight, and premature delivery (p < 0.05). The neonates from Guiyu are exposed to high levels of PBDEs. Prenatal exposure to PBDEs may affect neonates' health in Guiyu, which still needs to be evaluated in larger epidemiological studies. PMID- 20000819 TI - Electrical manipulation of spin states in a single electrostatically gated transition-metal complex. AB - We demonstrate an electrically controlled high-spin (S = 5/2) to low-spin (S = 1/2) transition in a three-terminal device incorporating a single Mn(2+) ion coordinated by two terpyridine ligands. By adjusting the gate-voltage we reduce the terpyridine moiety and thereby strengthen the ligand-field on the Mn-atom. Adding a single electron thus stabilizes the low-spin configuration and the corresponding sequential tunnelling current is suppressed by spin-blockade. From low-temperature inelastic cotunneling spectroscopy, we infer the magnetic excitation spectrum of the molecule and uncover also a strongly gate-dependent singlet-triplet splitting on the low-spin side. The measured bias-spectroscopy is shown to be consistent with an exact diagonalization of the Mn-complex, and an interpretation of the data is given in terms of a simplified effective model. PMID- 20000820 TI - Adsorption and conformation of a cationic surfactant on single-walled carbon nanotubes and their influence on naphthalene sorption. AB - Surfactants are used in synthesis and dispersion of carbon nanotubes (CNTs) and can thus be released into the environment with CNTs. In this study, it was observed that the coupled release of surfactants and CNTs altered the sorption of organic contaminants on the CNTs. The cationic surfactant, cetylpyridinium chloride (CPC), decreased naphthalene sorption on single-walled carbon nanotubes (SWCNT). In the most dramatic example, the adsorption capacity of naphthalene on SWCNT was reduced from 240 to 61.1 mg/g. The decrease of naphthalene sorption could be largely attributed to the competition of adsorbed CPC cations (i.e., C(21)H(38)N(+)) with naphthalene by occupying the SWCNT surface as surfaces decreased from 737 to 88.9 m(2)/g after the coating of CPC. However, the adsorbed CPC may form hemimicelles and result in a favorable media for naphthalene partition to counteract the decrease in naphthalene sorption. Configuration changes of adsorbed CPC hemimicelles might occur because the naphthalene partition into the adsorbed CPC decreased with the increase of adsorbed CPC. A partition-adsorption model was introduced to describe the partition fraction of naphthalene into adsorbed CPC hemimicelles as well as the adsorption fraction of naphthalene on unoccupied surfaces of SWCNT. PMID- 20000821 TI - Localizing and tracking single nanoscale emitters in three dimensions with high spatiotemporal resolution using a double-helix point spread function. AB - Three-dimensional nanoscale localization and tracking of dim single emitters can be obtained with a widefield fluorescence microscope exhibiting a double-helix point spread function (DH-PSF). We describe in detail how the localization precision quantitatively depends upon the number of photons detected and the z position of the nanoscale emitter, thereby showing a approximately 10 nm localization capability along x, y, and z in the limit of weak emitters. Experimental measurements are compared to Fisher information calculations of the ultimate localization precision inherent in the DH-PSF. The DH-PSF, for the first time, is used to track single quantum dots in aqueous solution and a quantum dot labeled structure inside a living cell in three dimensions. PMID- 20000822 TI - Behavior of decabromodiphenyl ether (BDE-209) in the soil-plant system: uptake, translocation, and metabolism in plants and dissipation in soil. AB - Deca-bromodiphenyl ether (BDE-209) is the major component of the commercial deca BDE flame retardant. There is increasing concern over BDE-209 due to its increasing occurrence in the environment and in humans. In this study the behavior of BDE-209 in the soil-plant system was investigated. Accumulation of BDE-209 was observed in the roots and shoots of all the six plant species examined, namely ryegrass, alfalfa, pumpkin, summer squash, maize, and radish. Root uptake of BDE-209 was positively correlated with root lipid content (P < 0.001, R(2) = 0.81). The translocation factor (TF, C(shoot)/C(root)) of BDE-209 was inversely related to its concentration in roots. Nineteen lower brominated (di- to nona-) PBDEs were detected in the soil and plant samples and five hydroxylated congeners were detected in the plant samples, indicating debromination and hydroxylation of BDE-209 in the soil-plant system. Evidence of a relatively higher proportion of penta- through di-BDE congeners in plant tissues than in the soil indicates that there is further debromination of PBDEs within plants or low brominated PBDEs are more readily taken up by plants. A significant negative correlation between the residual BDE-209 concentration in soil and the soil microbial biomass measured as the total phospholipid fatty acids (PLFAs) (P < 0.05, R(2) = 0.74) suggests that microbial metabolism and degradation contribute to BDE-209 dissipation in soil. These results provide important information about the behavior of BDE-209 in the soil-plant system. PMID- 20000823 TI - Effects of bioconjugation on the structures and electronic spectra of CdSe: density functional theory study of CdSe-adenine complexes. AB - We present density functional theory (DFT) and time-dependent DFT (TD-DFT) study of the structures and electronic spectra of small CdSe nanocluster-adenine complexes Cd(n)Se(n)-adenine (n = 3, 6, 10, 13). We examine the changes in the geometries and excitation spectra of the nanoclusters induced by DNA base binding. By comparing the results calculated for the bare (Cd(n)Se(n)), hydrogen passivated (Cd(n)Se(n)H(2n)), as well as the corresponding adenine (Ade)-bound clusters (Cd(n)Se(n)-Ade, Cd(n)Se(n)H(2n)-Ade, Cd(n)Se(n)H(2n-2)-Ade), we find that binding with Ade slightly blue-shifts (up to 0.18 eV) the electronic excitations of bare nanoclusters but strongly red-shifts (<1.2 eV) those of hydrogen-passivated nanoclusters. Natural bond orbital analysis shows that the LUMO of Cd(n)Se(n)H(2n)-Ade is a pi* orbital located on the purine ring. PMID- 20000824 TI - The network of receptors characterize B cell receptor micro- and macroclustering in a Monte Carlo model. AB - During the recognition of soluble antigens, B cell receptors (BCR) are known to form signaling clusters that can crucially modulate intracellular activation pathways and B cell response. Little is known about the precise nature of receptor cluster and its formation mechanism for the case of soluble antigens. Initial experiments have shown that B cell receptors first microcluster upon ligation with soluble antigens, and then coarsen into a macroscopic cap structure at one pole of a B cell. Such a mutual receptor-receptor attraction can arise locally due to cross-linking by soluble antigens among other possibilities. We develop an energy based Monte Carlo model to investigate the mechanism of B-cell receptor clustering upon ligation with soluble antigens. Our results show that mutual attraction between nearest neighbor receptor pairs can lead to microclustering of B cell receptors, but it is not sufficient for receptor macroclustering. A simple model of biased diffusion where BCR molecules experience a biased directed motion toward the largest cluster is then applied, which results in a single macrocluster of receptor molecules. The various types of receptor clusters are analyzed using the developed network-based metrics such as the average distance between any pairs of receptors. PMID- 20000825 TI - trans-Pyridyl and naphthyridyl cinnamides as alternatives for urea in complexation of carboxylic acid and formation of water-templated assemblies in the solid state. AB - A series of alpha,beta-unsaturated amides of pyridine and naphthyridine (1-4) derived from trans-cinnamic acid and its derivatives has been synthesized. The hydrogen-bonding behavior of these simple amides in the solid state has been established by solving X-ray structures. In solution and in the solid state, the above compounds 1-4 were found to bind carboxylic acid utilizing both conventional and unconventional hydrogen bonds. Hence, the possibility has been revealed of using this motif as an alternative of urea for the design of task specific hosts. The experimental results have been rationalized with an accompanying theoretical study involving density functional calculations on the structure and the relative energies of different rotamers of the host systems as well as the binding energy of the complexes formed with trans-cinnamic acid. Generally, small binding energies (0.58-0.70 eV) are calculated for the complexes in the gas phase which become even smaller in chloroform solvent. PMID- 20000826 TI - Dynamic solvent effects in electrochemical kinetics: indications for a switch of the relevant solvent mode. AB - The influence of solvent dielectric relaxation on the rate of electron transfer (ET) at an electrochemical interface is addressed using both experiment and model calculations. Water-ethylene glycol (EG) mixtures were chosen as the solvent because their optical permittivity remains practically constant over the entire composition range. This allows observation of the dynamic solvent effect with a very minor interference from the static solvent properties (being typically of opposite sign). Three groups of experimental results are presented to characterize the mixed-solvent system (dielectric spectra in the frequency range 0.1-89 GHz), the mercury/solvent interface (electrocapillary data), and the ET kinetics (dc polarography of peroxodisulphate reduction). To extract the true solvent influence on the electron transfer elementary step, the results from dc polarography are corrected for interfacial effects with the help of the electrocapillary data. An anomalous dependence of the ET rate on EG content (i.e., nonmonotonic dependence of the ET rate on macroscopic viscosity) can be inferred after all corrections. The interplay of different solvent modes is suggested to be responsible for the observed features of ET kinetics. A possible interpretation of the corrected ET rate in the framework of the Agmon-Hopfield formalism is proposed, where the dielectric spectra of the mixed solvent are modeled by a superposition of three Debye equations. The results demonstrate that the observed anomalous "viscosity effect" may be explained qualitatively by an increased contribution of the fast relaxation mode at high EG contents. PMID- 20000827 TI - Patterned electroconvective states in a bent-core nematic liquid crystal. AB - We report the results of investigations on the anisotropic electrohydrodynamic states arising in a highly conducting, planarly aligned, bent-core nematic liquid crystal driven by ac fields of frequency f in the range from 10 Hz to 1 MHz. Pattern morphologywise, two f regimes are distinguished. The low-f regime, wherein the primary bifurcation is to a state of periodic longitudinal stripes (LS), extends to an unprecedentedly large f, in the range 150-550 kHz, depending on the temperature T. This is followed by the high-f regime wherein periodic normal stripes (NS) constitute the primary instability. Both instabilities involve predominant director modulations and streamlines in the layer plane. The transitional frequency between the two regimes is linear in temperature. The curve V(c)(f) shows a nonlinear increase for the LS state and decrease for the NS state. V(c)(T) is an ever increasing curve close to the nematic-isotropic point for both states. The wavenumber of LS varies directly as V, and that of NS shows nearly the same behavior. The pattern period versus f is increasing for LS but decreasing for NS. Both instability states exhibit complex, light-polarization dependent lens action. Well above the threshold, disclination loops of regular geometry appear along the stripes. They drift in a coordinated manner along the flow lines. At very high voltages, the instability turns strongly time dependent. The current models of anisotropic convection based on static electrical parameters fail to account for the observed instabilities. PMID- 20000828 TI - Frustrated phase transformations in supported, interdigitating lipid bilayers. AB - In free bilayers, the fluid to gel main phase transition of a monofluorinated phospholipid (F-DPPC) transforms a disordered fluid bilayer into a fully interdigitated monolayer consisting of ordered acyl tails. This transformation results in an increase in molecular area and decrease in bilayer thickness. We show that when confined in patches near a solid surface this reorganization proceeds under constraints of planar topography and total surface area. One consequence of these constraints is to limit the complete formation of the energetically favored, interdigitated gel phase. The noninterdigitated lipids experience enhanced lateral tension, due to the expansion of the growing interdigitated phase within the constant area. The corresponding rise in equilibrium transition temperatures produces supercooled lipids that vitrify when cooled further. Ultimately, this frustrated phase change reflects a coupling between dynamics and thermodynamics and gives rise to an unusual phase coexistence characterized by the presence of two qualitatively different gel phases. PMID- 20000829 TI - Bayesian detection of intensity changes in single molecule and molecular dynamics trajectories. AB - Single molecule spectroscopy experiments and molecular dynamics simulations have several profound features in common, chief among which is that both follow the dynamics of some degrees of freedom of a single molecule over time. The analysis is essentially the same: one investigates the changes in the degrees of freedom followed. For instance, in a single molecule fluorescence experiment, the degree of freedom is often the number of photons detected in some time period. In this article, we introduce a straightforward Bayesian method for detecting if and when changes occurred. In contrast to methods based upon maximum likelihood estimates, a Bayesian approach allows for a more systematic means not only to change point detection but also to cluster the data into states. Most importantly, the Bayesian method supplies a simpler hypothesis testing framework. Although we focus on Poisson-distributed data, the Bayesian methods outlined here can in principle be applied to data sampled from any distribution. PMID- 20000830 TI - Fe2(SO4)3 as a binary oxidant and dopant to thin polyaniline nanowires with high conductivity. AB - This article exposes a facial approach to self-assemble polyaniline (PANI) nanowires with thin diameter (approximately 10 nm) and high room-temperature conductivity (approximately 10(0) S/cm) by using Fe(2)(SO(4))(3) as a binary oxidant and dopant. The new method not only saves hard templates and postprocess of template removal but also simplifies the reagent. Formation yield, diameter, and room-temperature conductivity of the nanowires are affected by the molar ratios of Fe(2)(SO(4))(3) to aniline. The low redox potential of Fe(2)(SO(4))(3) not only results in a thinner diameter and higher room-temperature conductivity (10(0) S/cm) of the nanowires but also shows a much weaker temperature dependence of resistivity and smaller characteristic Mott temperature (T(0) = 2.5 x 10(3) K). PMID- 20000831 TI - Excited-state properties of octahedral hexarhenium(III) complexes with redox active N-heteroaromatic ligands. AB - New sulfide-capped octahedral hexarhenium(III) complexes containing 4 phenylpyridine (ppy) or 1,2-bis(4-pyridyl)ethane (bpe) ((n-C(4)H(9))(4)N)[mer {Re(6)S(8)Cl(3)(ppy)(3)}] ((Bu(4)N)[1]), ((n-C(4)H(9))(4)N)(2)[trans {Re(6)S(8)Cl(4)(ppy)(2)}] ((Bu(4)N)(2)[2a]), ((n-C(4)H(9))(4)N)(2)[cis {Re(6)S(8)Cl(4)(ppy)(2)}] ((Bu(4)N)(2)[2b]), ((n-C(4)H(9))(4)N)(2)[trans {Re(6)S(8)Cl(4)(bpe)(2)}] ((Bu(4)N)(2)[3a]), and ((n-C(4)H(9))(4)N)(2)[cis {Re(6)S(8)Cl(4)(bpe)(2)}] ((Bu(4)N)(2)[3b]) were prepared, and X-ray single crystal structure determination was carried out for (Bu(4)N)(2)[2a] and (Bu(4)N)(2)[3a]. The photophysical properties of these complexes were studied both in acetonitrile at 298 K and in the solid state at 298 and 80 K, along with those of the known 4,4'-bipyridine (bpy) analogues ((n-C(4)H(9))(4)N)[mer {Re(6)S(8)Cl(3)(bpy)(3)}] ((Bu(4)N)[4]), ((n-C(4)H(9))(4)N)(2)[trans {Re(6)S(8)Cl(4)(bpy)(2)}] ((Bu(4)N)(2)[5a]), and ((n-C(4)H(9))(4)N)(2)[cis {Re(6)S(8)Cl(4)(bpy)(2)}] ((Bu(4)N)(2)[5b]). The photophysical data of [5a](2-) and [5b](2-) in solution and in the solid state were significantly different from those of other complexes. On the basis of experimental observations of [2a](2-) and [5a](2-) and density-functional theory (DFT) calculations, it was concluded that [5a](2-) and [5b](2-) exhibited metal (Re(6)S(8) core)-to-ligand (bpy) charge transfer (MLCT) type emission. This is the first unambiguous demonstration of MLCT type emissions for the hexarhenium complexes. The MLCT components, where present, are only minor in the case of the emissions of [1](-), [2a](2-), [2b](2 ), and [4](-); these can be explained primarily as the contributions of the intracore electronic transitions. The emissions of [3a](2-) and [3b](2-) can be assigned almost completely to the electronic transitions within the Re(6)S(8) core. The different emission characteristics of the bis(bpy) complexes ([5a](2-) and [5b](2-)) from the tris(bpy) complex ([4](-)) are a result of the increase in the number of nitrogen donors on the Re(6)S(8) core, which stabilizes the Re(6)S(8) core energy to a lower level than the energy of the bpy ligand pi* orbital. On the other hand, it has been shown that the emissions of the bis(ppy) ([2a](2-) and [2b](2-)) and bis(bpe) complexes ([3a](2-) and [3b](2-)) are best characterized by the higher pi* energy level of each N-heteroaromatic ligand, which lead to a stronger metal character in the emissive excited state of the complex. PMID- 20000832 TI - Heteroatomic deltahedral zintl ions of group 14 and their alkenylation. AB - Reported is the synthesis of Ge(9-x)Sn(x) heteroatomic deltahedral Zintl ions and their alkenylation by reactions with alkynes. The nine-atom clusters are made either by extraction from mixed Ge/Sn precursors with nominal composition K(4)Ge(9-x)Sn(x) or by dissolution of mixtures of the corresponding binary precursors K(4)Ge(9) and K(4)Sn(9) in solvents with high dielectric constants such as DMF, DMSO, and acetonitrile. Reactions of the heteroatomic clusters with alkynes such as Me(3)SiC[triple bond]CSiMe(3), HC[triple bond]CCpr (Cpr = cyclopropyl), and HC[triple bond]CPh in ethylenediamine resulted in the following structurally characterized compounds with alkenylated heteroatomic clusters: [K (2,2,2-crypt)](3)[GeSn(8)-CH=CH(2)].en.tol (1), triclinic, P1, a = 13.9220(3) A, b = 14.9788(3) A, and c = 21.5892(5) A, alpha = 94.2580(10) degrees , beta = 98.5210(10) degrees , and gamma = 98.4890(10) degrees , V = 4382.31(16) A(3), Z = 2; [K-(2,2,2-crypt)](4)[Ge(2)Sn(7)(CH=CH(2))(2)](2).en (2), monoclinic, P2(1)/c, a = 48.1883(15) A, b = 12.1551(4) A, and c = 21.4824(7) A, beta = 90.052(2) degrees , V = 12583.0(7) A(3), Z = 4; [K-(2,2,2-crypt)](3)[GeSn(8)-CH=CHCpr].en (3), monoclinic, P2(1)/c, a = 17.9132(9) A, b = 22.7967(11) A, and c = 21.6922(12) A, beta = 98.409(2) degrees , V = 8763.0(8) A(3), Z = 4; [K-(2,2,2 crypt)](3)[Ge(2)Sn(7)-CH=CHPh].2en (4), monoclinic, P2(1)/n, a = 13.2583(5) A, b = 47.0565(17) A, and c = 15.9978(6) A, beta = 111.536(2) degrees , V = 9284.1(6) A(3), Z = 4. The potassium countercations of the divinyl-substituted cluster in 2 were exchanged for tetrapropylammonium cations, and the resulting compound was also crystallized and structurally characterized: [Pr(4)N](4)[Ge(2)Sn(7)(CH=CH(2))(2)](2) (5), triclinic, P1, a = 11.6757(8) A, b = 18.8150(16) A, and c = 21.0608(17) A, alpha = 112.327(3) degrees , beta = 91.550(3) degrees , and gamma = 91.892(3) degrees , V = 4273.5(6) A(3), Z = 2. All clusters were also characterized in solution by electrospray mass spectrometry. PMID- 20000833 TI - Steric titration of arylthiolate coordination modes at pseudotetrahedral nickel(II) centers. AB - Several derivatives of the pseudotetrahedral phenylthiolate complex Tp(Me,Me)Ni SPh (1), Tp(Me,Me-) = hydrotris(3,5-dimethyl-1-pyrazolyl)borate, were prepared incorporating substituted arylthiolates, including a series of ortho-substituted ligands Tp(Me,Me)Ni-SR (R = 2,6-Me(2)C(6)H(3), 2; 2,4,6-Me(3)C(6)H(2), 3; 2,4,6- (i)Pr(3)C(6)H(2), 4; and 2,6-Ph(2)C(6)H(3), 5) and a series of para-substituted complexes (R = C(6)H(4)-4-OMe, 6; C(6)H(4)-4-Me, 7; and C(6)H(4)-4-Cl, 8). The products were characterized by (1)H NMR and UV-vis spectroscopy. Spectra of 6-8 were consistent with retention of a common structure across the para-substituted series with modest perturbation of the spectral features of 1 assisting their assignment. In contrast, spectra of 2-5 were indicative of a significant change in configuration across the ortho-disubstituted series. The structure of complex 5 was determined by X-ray crystallography and a distinctive arylthiolate ligation mode was found, in which the N(3)S ligand field was significantly distorted toward a sawhorse, compared to a more common trigonal pyramidal shape (e.g., 1). Moreover, the arylthiolate substituent rotated from a vertical orientation co directional with the pyrazole rings and disposed between two of them in 1, to a horizontal orientation perpendicular to and over a single pyrazole ring in 5. This reorientation is necessary to accommodate the large ortho substituents of the latter complex. The divergent Ni-S coordination modes result in distinct (1)H NMR and electronic spectra that were rationalized by density functional theory (DFT) and time-dependent DFT (TD-DFT) calculations. These results demonstrate rich coordination chemistry for arylthiolates that can be elicited by steric manipulation at the periphery of pseudotetrahedral ligand fields. PMID- 20000834 TI - Magnetism and structure in chains of copper dinuclear paddlewheel units. AB - An anhydrous copper carboxylate compound of formula [Cu(trans-2-butenoate)(2)](n) has been characterized. X-ray analysis reveals a structure built by paddlewheel units bridged by pairs of Cu...O axial bonds to give infinite chains arranged in a new topological motif. Susceptibility measurements in the 10-300 K temperature range, and isothermal magnetization curves at 2, 5, 10, and 50 K with fields up to 5 T, were obtained. Electron Paramagnetic Resonance (EPR) spectra of powder samples were measured at 33.9 GHz at 300 K, and at 9.60 GHz at temperatures in the range 90 2 > 1, suggesting that the arene plays an important role in the biological activity of these types of compounds. Cell distribution studies using fractionation showed that all four compounds distribute similarly within cells. DNA binding of osmium did not correlate with cytotoxicity, indicating that the nature of the DNA lesion may also be crucial to activity. TEM images of ovarian cells treated with 3 revealed morphological changes associated with apoptosis with possible involvement of mitochondria. PMID- 20000848 TI - On-chip activation and subsequent detection of individual antigen-specific T cells. AB - The frequencies of antigen-specific CD4+ T cells in samples of human tissue have been difficult to determine accurately ex vivo, particularly for autoimmune diseases such as multiple sclerosis or type 1 diabetes. Conventional approaches involve the expansion of primary T cells in vitro to increase the numbers of cells, and a subsequent assessment of the frequencies of antigen-specific T cells in the expanded population by limiting dilution or by using fluorescently labeled tetramers of peptide-loaded major histocompatibility complex (MHC) receptors. Here we describe an alternative approach that uses arrays of subnanoliter wells coated with recombinant peptide-loaded MHC class II monomers to isolate and stimulate individual CD4+ T cells in an antigen-specific manner. In these experiments, activation was monitored using microengraving to capture two cytokines (IFNgamma and IL-17) released from single cells. This new method should enable direct enumeration of antigen-specific CD4+ T cells ex vivo from clinical samples. PMID- 20000849 TI - Factorial Kriging Analysis as a tool for explaining the complex spatial distribution of metals in sediments. AB - Rivers flowing through urbanized and industrial areas are usually greatly damaged by anthropogenic activities discharging contaminants. Characterizing the spatial distribution of pollutants in sediments is of high importance for selecting a suitable remediation operation, but is a complex task because this spatial variability is the result of various physical and chemical mechanisms occurring at different scales. Factorial Kriging Analysis (FKA) was applied on data collected in a canalized river (Scarpe, France) for that purpose, because this geostatistical technique allows to decompose a given variable into components of different spatial correlations and map them separately. This decomposition is meaningful provided that it can be related to physical phenomena occurring at the identified spatial scales. FKA applied to Cd and Zn concentrations in sediments of the Scarpe river proved to be effective, allowing their mapping to be decomposed in a first map related to a short-range spatial correlation corresponding to hot spots interpreted as the impact of industrial and urban inputs located along the canal, and a second map related to a long-range spatial variability associated with long pollutant plumes interpreted as the effect of one major upstream pollutant input. PMID- 20000850 TI - Carboxylation of Kinetically Inert Platinum(IV) Hydroxy Complexes. An Entr.acte.ee into Orally Active Platinum(IV) Antitumor Agents. PMID- 20000851 TI - Structural insights into substrate specificity and the anti beta-elimination mechanism of pectate lyase. AB - Pectate lyases harness anti beta-elimination chemistry to cleave the alpha-1,4 linkage in the homogalacturonan region of plant cell wall pectin. We have studied the binding of five pectic oligosaccharides to Bacillus subtilis pectate lyase in crystals of the inactive enzyme in which the catalytic base is substituted with alanine (R279A). We discover that the three central subsites (-1, +1, and +2) have a profound preference for galacturonate but that the distal subsites can accommodate methylated galacturonate. It is reasonable to assume therefore that pectate lyase can cleave pectin with three consecutive galacturonate residues. The enzyme in the absence of substrate binds a single calcium ion, and we show that two additional calcium ions bind between enzyme and substrate carboxylates occupying the +1 subsite in the Michaelis complex. The substrate binds less intimately to the enzyme in a complex made with a catalytic base in place but in the absence of the calcium ions and an adjacent lysine. In this complex, the catalytic base is correctly positioned to abstract the C5 proton, but there are no calcium ions binding the carboxylate at the +1 subsite. It is clear, therefore, that the catalytic calcium ions and adjacent lysine promote catalysis by acidifying the alpha-proton, facilitating its abstraction by the base. There is also clear evidence that binding distorts the relaxed 2(1) or 3(1) helical conformation of the oligosaccharides in the region of the scissile bond. PMID- 20000852 TI - Effects of different rearing and feeding systems on lipid oxidation and antioxidant capacity of freeze-dried egg yolks. AB - Lipid oxidation and antioxidant capacity of freeze-dried egg yolks produced with two rearing systems (battery cages and free-range) and two types of feedings (conventional and organic) were studied. Nine fresh egg yolks of each crossed treatment were pooled, frozen for a month, freeze-dried, vacuum-packed, and kept at -18 degrees C until analysis. No significant differences were observed in the lipid (58.0-62.1%) and total sterol contents (33.0-35.5 g/kg of lipids) of the freeze-dried egg yolks. Free rearing and conventional feeding systems resulted in significantly higher total tocopherol, alpha-tocopherol, and lutein contents, as compared to the battery cage and the organic feed, respectively. However, no significant differences were found in lipid oxidation (peroxide value = 0.7-0.9 mequiv of O(2)/kg of fat; thiobarbituric reactive substances = 1.0-1.3 mg of malonylaldehyde/kg of sample) and cholesterol oxidation (28.8-43.5 mg of cholesterol oxidation products/kg of lipids; 0.08-0.12% oxidized cholesterol) of freeze-dried egg yolks except for 7alpha-hydroxycholesterol, which was significantly lower in samples obtained with organic feed. PMID- 20000853 TI - Characteristic aroma-active compounds of Korean perilla (Perilla frutescens Britton) leaf. AB - Aroma-active compounds from Korean perilla (Perilla frutescens Britton) leaf were extracted by solvent-assisted flavor evaporation (SAFE), liquid-liquid continuous extraction (LLCE), and hydrodistillation (HD) and analyzed by gas chromatography mass spectrometry (GC-MS) and gas chromatography-olfactometry (GC-O). Thirty three volatile compounds were identified by GC-MS. 1-(3-Furyl)-4-methyl-1 pentanone (perilla ketone) was found to be the most abundant volatile compound, followed in order by (Z)-3-hexenol and 1-octen-3-ol. Perilla ketone comprised 81% (93 ppm), 84% (120 ppm), and 95% (490 ppm) of the volatile compounds obtained from SAFE, LLCE, and HD, respectively. Thirteen aroma-active compounds were detected by GC-O. Perilla ketone, 1-(3-furyl)-4-methyl-3-penten-1-one (egoma ketone), and 1-(3-furyl)-4-methyl-2-penten-1-one (isoegoma ketone) were considered to be the characteristic aroma-active compounds of Korean perilla leaf. Perilla ketone, (Z)-3-hexenal (green), egoma ketone, and isoegoma ketone were the most intense aroma-active compounds in Korean perilla leaf. Other relatively intense odorants included (Z)-3-hexenol (green), (E)-2-hexenal (green), benzaldehyde (almond), 1-octen-3-one (metallic), 1-octen-3-ol (mushroom), phenylacetaldehyde (honeysuckle), linalool (lemon), and beta caryophyllene (woody). PMID- 20000854 TI - Organocatalysis in organic synthesis. PMID- 20000855 TI - New light on molecular and materials complexity: 4D electron imaging. AB - In this Perspective, 4D electron imaging is highlighted, after introducing some concepts, with an overview of selected applications that span chemical reactions, molecular interfaces, phase transitions, and nano(micro)mechanical systems. With the added dimension of time in microscopy, diffraction, and electron-energy-loss spectroscopy, the focus is on direct visualization of structural dynamics with atomic and nanoscale resolution in the four dimensions of space and time. This contribution provides an expose of emerging developments and an outlook on future applications in materials and biological sciences. PMID- 20000856 TI - A bimetallic Ru2Pt complex containing a trigonal-planar mu3-carbido ligand: formation, structure, and reactivity relevant to the Fischer-Tropsch process. AB - The bimetallic Ru(2)Pt complex [(Cp*Ru)(2)(mu(2)-NHPh)(mu(2)-H)(mu(3) C)PtMe(PMe(3))(2)][OTf] (3; Cp* = eta(5)-C(5)Me(5)) containing a planar three coordinate carbido ligand has been prepared in 93% yield by thermal isomerization of the bridging methylene precursor [(Cp*Ru)(2)Me(mu(3)-NPh)(mu(2) CH(2))Pt(PMe(3))(2)][OTf] (2) via cleavage of the methylene C-H bonds. Exposure of the carbido complex 3 to carbon monoxide (1 atm) induced coupling of the carbido ligand with the nearby methyl and hydride ligands to produce the diruthenium ethylidene complex [(Cp*Ru)(2)(mu(2)-CHMe)(mu(2)-NHPh)(CO)(2)][OTf] (4) and the known triplatinum complex [Pt(CO)(PMe(3))](3). The crystal structures of 2, 3, and 4 (BPh(4) salt) are reported. PMID- 20000857 TI - N-heterocyclic carbene-catalyzed enantioselective Mannich reactions with alpha aryloxyacetaldehydes. AB - N-Heterocyclic carbenes (NHCs) catalyze a new Mannich-type reaction to form beta amino acid derivatives in high yield and enantioselectivity. The reaction is initiated by the addition of an NHC to an alpha-aryloxyaldehyde followed by elimination of a phenoxide anion which generates an enol/enolate. A Mannich addition to a tosylimine proceeds with excellent control over enantioselectivity. In a new carbene catalysis concept, catalyst regeneration is promoted by return, or rebound, and acylation of the phenoxide group which served as the activating component in the first step of the catalytic cycle. The activated ester products formed in situ are manipulated to form a variety of useful compounds including beta-amino acids, beta-amino amides, and peptides. PMID- 20000858 TI - Synthesis of dialkyl ethers from organotrifluoroborates and acetals. AB - The formation of ethers by C-O bond formation under harsh basic or acidic conditions is an entrenched synthetic disconnection in organic chemistry. We report a strategic alternative that involves the BF(3).OEt(2)-promoted coupling of stable, easily prepared acetals with widely available potassium aryl-, alkenyl , and alkynyltrifluoroborates. This fast, operationally simple process offers straightforward access to dialkyl ethers, many of which would be difficult to prepare using classical methods. The use of MOM-protected alcohols and acetal protected aldehydes enables ether formation without recourse to protecting-group manipulations or strong bases. PMID- 20000859 TI - Direct inhibition of hypoxia-inducible transcription factor complex with designed dimeric epidithiodiketopiperazine. AB - Selective blockade of hypoxia-inducible gene expression by designed small molecules would prove valuable in suppressing tumor angiogenesis, metastasis and altered energy metabolism. We report the design, synthesis, and biological evaluation of a dimeric epidithiodiketopiperazine (ETP) small molecule transcriptional antagonist targeting the interaction of the p300/CBP coactivator with the transcription factor HIF-1alpha. Our results indicate that disrupting this interaction results in rapid downregulation of hypoxia-inducible genes critical for cancer progression. The observed effects are compound-specific and dose-dependent. Controlling gene expression with designed small molecules targeting the transcription factor-coactivator interface may represent a new approach for arresting tumor growth. PMID- 20000860 TI - Nitric oxide oxidatively nitrosylates Ni(I) and Cu(I) C-organonitroso adducts. AB - Monovalent nickel and copper beta-diketiminato complexes react with ArN=O (Ar = 3,5-Me(2)C(6)H(3), Ph) to give C-nitroso adducts that exhibit three different modes of bonding with varying degrees of N-O bond activation. The addition of ArNO to 2 equiv of [Me(2)NN]Ni(2,4-lutidine) {[Me(2)NN](-) = 2,4-bis(2,6 dimethylphenylimido)pentyl} gives {[Me(2)NN]Ni}(2)(mu-eta(2):eta(2)-ONAr) (1a and 1b), which exhibit symmetrical bonding of the ArN=O moiety between two [Me(2)NN]Ni fragments, with a N-O bond distance of 1.440(4) A in 1a that is significantly longer than those in free C-organonitroso compounds (1.13-1.29 A). [Me(2)NN]Cu(NCMe) reacts with 0.5 equiv of ArNO in ether to give the dinuclear adducts {[Me(2)NN]Cu}(2)(mu-eta(2):eta(1)-ONAr) (2a and 2b), which exhibit eta(2) and eta(1) bonding of the ArN=O moiety with separate [Me(2)NN]Cu fragments possessing N-O distances of 1.375(6) A (2a) and 1.368(2) A (2b). In arene solvents, one beta-diketiminatocopper(I) fragment dissociates from 2 to give [Me(2)NN]Cu(eta(2)-ONAr) (3a and 3b), which may be isolated by the addition of 1 equiv of ArNO to [Me(2)NN]Cu(NCMe). The X-ray structures of 3a and 3b are similar to those of related Cu(I) alkene adducts, with N-O distances in the narrow range 1.333(4)-1.338(5) A. IR spectra of the nitrosobenzene adducts 1b, 2b, and 3b exhibit nu(NO) stretching frequencies at 915, 1040, and 1113 cm(-1), respectively, following the decreasing degree of N=O activation observed in the X ray structures of species 1, 2, and 3. Both 1a and 3a react with anaerobic NO(g) to give the corresponding N-aryl-N-nitrosohydroxylaminato complexes [Me(2)NN]M(kappa(2)-O(2)N(2)Ar) [M = Ni (4), Cu (5)]. In the reaction of dinuclear 1a with NO, one [Me(2)NN]Ni fragment is trapped as the nickel nitrosyl [Me(2)NN]Ni(NO). Reaction of the monovalent complex [Me(2)NN]Cu(eta(2)-ONAr) with NO(g) to give divalent [Me(2)NN]Cu(kappa(2)-O(2)N(2)Ar) represents an example of oxidative nitrosylation. PMID- 20000861 TI - Designed surface with tunable IgG density as an in vitro model for immune complex mediated stimulation of leukocytes. AB - We present the design of an in vitro model for immune-complex-mediated stimulation of leukocytes and its functional characteristics with respect to monocyte adhesion. The model was based on the orientation-controlled immobilization of a humanized IgG1 monoclonal antibody (rituximab) via its interaction with a biotinylated peptide epitope derived from the CD20 marker. The peptide was linked to neutravidin covalently attached to a mixed self-assembled monolayer of carboxyl- and methoxy-terminated oligo(ethylene glycol) alkane thiolates on gold. The surface adhesion propensity of human monocytes (cell line U937) was highly dependent on the lateral IgG density and indicated that there exists a distance between IgG-Fc on the surface where interactions with Fc gamma receptors are optimal. This well-defined platform allows for a careful control of the size and orientation of artificial IgG immune complexes, it is easily made compatible with, for example, cellular imaging, and it will become useful for in vitro studies on the importance of Fc gamma receptor interactions in chronic immune-mediated diseases. PMID- 20000862 TI - Thorough QT Studies: Questions and Quandaries. AB - The International Conference on Harmonisation E14 Guidance was successful in largely standardizing the conduct of the so-called thorough QT/QTc studies (TQTS). Nevertheless, there is still a spectrum of frequently encountered problems with details of design, conduct and interpretation of TQTS. Several of these challenges are reviewed here, starting with explaining that the TQTS goal is only to identify drugs for which the proarrhythmic risk might be considered excluded for the purposes of regulatory benefit-risk assessment. Suggestions are made on how to categorize and quantify or exclude proarrhythmic risk if the TQTS is positive. There is a conceptual need for TQTS, and this is discussed, together with reasons why restricted clinical registries cannot prove the absence of proarrhythmic liability of any drug. Appropriate drug doses investigated in TQTS should be derived from the maximum clinically tolerable dose rather than from the known or expected therapeutic dose. With the help of concentration-QTc modelling, the standard therapeutic dose can be omitted from TQTS, especially if the study is expected to be negative. Conditions for single-dose TQTS acceptability are reviewed. The role of the so-called positive control is assessed, contrasting the role of a same-class comparator for the investigated drug. A single 400 mg dose of moxifloxacin is advocated as the present 'gold standard' assay sensitivity test. The necessity of careful placebo control is explained and the frequency of ECG assessments is considered. The central tendency and outlier analyses are discussed, together with the correct approaches to baseline adjustment. The review concludes that the design and interpretation of TQTS must not be approached with mechanistic stereotypes, and highlights the importance of relating the QTc changes to drug plasma levels. PMID- 20000863 TI - EIDOS: a mechanistic classification of adverse drug effects. AB - The mechanisms of adverse drug effects have not been adequately classified. Here, we propose a comprehensive mechanistic classification of adverse drug effects that considers five elements: the Extrinsic chemical species (E) that initiates the effect; the Intrinsic chemical species (I) that it affects; the Distribution (D) of these species in the body; the (physiological or pathological) Outcome (O); and the Sequela (S), which is the adverse effect. This classification, which we have called EIDOS, describes the mechanism by which an adverse effect occurs; it complements the DoTS classification of adverse effects (based on clinical pharmacology), which takes into account Dose responsiveness, Time course, and Susceptibility factors. Together, these two classification systems, mechanistic and clinical, comprehensively delineate all the important aspects of adverse drug reactions; they should contribute to areas such as drug development and regulation, pharmacovigilance, monitoring therapy, and the prevention, diagnosis, and treatment of adverse drug effects. PMID- 20000864 TI - Neuromuscular adverse effects associated with systemic retinoid dermatotherapy: monitoring and treatment algorithm for clinicians. AB - Although neuromuscular adverse effects represent significant clinical manifestations of hypervitaminosis A syndrome, surprisingly little attention has been paid to the potential neuromuscular toxicity of vitamin A derivatives (retinoids). Since isotretinoin and acitretin are currently the two most commonly used oral retinoids in systemic dermatotherapy, this review focuses exclusively on their neuromuscular adverse effects and proposes a neuromuscular algorithm for appropriate monitoring of patients treated with these two compounds. The most frequent CNS adverse effect associated with oral isotretinoin is headache, either as an independent adverse effect or as part of benign intracranial hypertension, which is additionally characterized by nausea and visual changes. Isolated cases of stiff-person-like syndrome, epileptic seizures and generalized muscle stiffness syndrome, possibly or probably related to oral treatment with isotretinoin, have also been reported. In addition, oral isotretinoin has reportedly been associated with muscular adverse effects that most frequently manifest as myalgia and stiffness and, in rare cases, as true myopathy or rhabdomyolysis. Creatine phosphokinase, a specific marker of muscle destruction, has been found to be elevated, occasionally by up to 100 times the normal value (with or without muscular symptoms and signs), in a variable percentage of patients receiving isotretinoin treatment and particularly in those undergoing vigorous physical exercise. Oral acitretin has been found to cause peripheral nerve dysfunction, particularly of sensory fibres, which in rare cases leads to clinically evident sensory disturbances. Less clear is the causal relationship between acitretin and benign intracranial hypertension or myopathy, whereas an isolated case of cranial nerve IV (oculomotor) palsy and a further case of thrombotic stroke during treatment with oral acitretin have been reported. Systemic diseases with involvement of nervous and/or muscle tissue and neuromuscular disorders should be regarded as exclusion criteria for initiation of oral retinoid therapy. Additionally, intense physical exercise and concurrent treatment with neurotoxic or myotoxic drugs should be avoided during treatment with oral retinoids. In order to minimize the potential risk of neuromuscular adverse effects, a neuromuscular algorithm is suggested that may be useful for monitoring patients taking oral retinoids. PMID- 20000865 TI - Resolving a double standard for risk management of thalidomide: an evaluation of two different risk management programmes in Japan. AB - Thalidomide, once withdrawn because of its teratogenicity, has now been re launched worldwide. In Japan, thalidomide has been imported by individual doctors since around the year 2000. In October 2008, it was approved for the treatment of multiple myeloma (MM) by the Ministry of Health, Labour and Welfare (MHLW) on the condition that the manufacturer implemented a risk management programme termed the Thalidomide Education and Risk Management System (TERMS). It is likely that the imports of thalidomide will be used off-label to treat diseases other than MM. Thus, the MHLW is also planning to introduce a web-based registration system, referred to as the Safety Management System for Unapproved Drugs (SMUD), for thalidomide imported by individual doctors. To evaluate the difference between TERMS and SMUD and establish a way to resolve the 'double standard' for risk management of thalidomide treatment in Japan. The fraction of patients with disorders other than MM was estimated by the volume of annual imports obtained from the MHLW and records of the imports for patients with MM, other oncological diseases (ODs) and non-ODs in 2007 through a major supplier covering 63% of the total imported thalidomide. The information for TERMS was obtained from web pages of the manufacturer and the MHLW. The components of TERMS were compared with those in SMUD. Provided that the distribution of the indication for thalidomide (MM) in 2007, estimated from the records of imports through the major supplier, is representative of the entire nation, it is estimated that on average 866 patients, including 851 (98.3%) with MM, are using thalidomide on any one day. However, if the major supplier's imports, which account for 63% of the total imports, are not representative of the nation as a whole, possibly only half of the patients treated with thalidomide in Japan have MM. This would be the case in a scenario where the remaining 37% of imports are exclusively used to treat disorders other than MM. TERMS consists of tools for education and registration of patients, and has the potential for real-time intervention. SMUD is a system for registration of patients and exchange of safety information between health professionals, but has some mandatory components that encourage patient registration. TERMS and SMUD are different in nature, and they impose different criteria that doctors and patients should satisfy in order to use thalidomide. To eliminate this double standard, implementation of a single system would be desirable. However, improvement of SMUD may be the second best option by developing tools for patient education, enhancing the potential for real-time intervention and monitoring thalidomide usage by each patient. On average, a total of about 1000 patients are estimated to be using thalidomide on any one day in Japan. It is likely that those patients are placed under one of two different risk management programmes. SMUD should be improved so that all patients are monitored in a way that results in a similar level of risk management. PMID- 20000866 TI - Syndrome of inappropriate antidiuretic hormone secretion (SIADH) or hyponatraemia associated with valproic Acid: four case reports from the Netherlands and a case/non-case analysis of vigibase. AB - The Netherlands Pharmacovigilance Centre Lareb received four cases of severe symptomatic hyponatraemia or syndrome of inappropriate antidiuretic hormone secretion (SIADH) in association with valproic acid use, in which a causal relationship was suspected. This study describes these cases and gives support for this association from Vigibase, the adverse drug reaction (ADR) database of the WHO Collaborating Centre for International Drug Monitoring, the Uppsala Monitoring Centre. Cases of hyponatraemia in valproic acid users are described. In a case/non-case analysis, the strength of the association between reported cases of hyponatraemia and the use of valproic acid in Vigibase was established by calculating a reporting odds ratio, adjusted for possible confounding by concomitant medication. Four females aged 57, 67, 71 and 88 years developed symptomatic hyponatraemia or SIADH after starting valproic acid. Despite concomitant medication or co-morbidity, a causal relationship was plausible. In Vigibase, valproic acid is disproportionally associated with hyponatraemia and SIADH (corrected reporting odds ratio 1.83 [95% CI 1.61, 2.08]). Based on the described cases and the reports from Vigibase, a causal relationship between valproic acid use and hyponatraemia or SIADH can be suspected. The mechanism by which valproic acid could cause hyponatraemia or SIADH has not been fully elucidated. Valproic acid use could lead to reduced sensitivity of hypothalamic osmoreceptors. It also might directly affect tubular cell function, thereby leading to SIADH. It might be expected that a combination of effects on the osmoreceptors and a lack of compensation of the salt-water unbalance by the nephrons causes SIADH in some patients using valproic acid. It could be a dose- or concentration-related adverse effect. In this report, severe symptomatic hyponatraemia and SIADH have been associated with the use of valproic acid. With this study, not only is the number of published cases doubled, but also the data from Vigibase strongly support the association. Since hyponatraemia and SIADH have a high morbidity, health professionals should be aware of this potential ADR. PMID- 20000867 TI - Diagnostic utility of two case definitions for anaphylaxis: a comparison using a retrospective case notes analysis in the UK. AB - Anaphylaxis is a clinical diagnosis with no gold-standard test. Recent case definitions have attempted to provide objective criteria for diagnosis. The aim of this study was to compare the diagnostic concordance of the Brighton Collaboration case definition (the 'Brighton' case definition) to the consensus case definition from the Second Symposium on the Definition and Management of Anaphylaxis (the 'Symposium' definition). The study setting was a hospital-based emergency department in the UK. We identified cases of anaphylaxis by physicians' discharge diagnoses over a 2-year period from 2005 to 2006, and used randomly selected cases of allergic reaction, asthma and urticaria as a control group. Data was extracted by clinicians (who were unaware of the content of either case definition), and the two case definitions were applied by Boolean operators in a Microsoft Excel spreadsheet. Concordance between the case definitions was measured using Cohen's kappa (kappa) statistic. We reviewed 128 sets of notes, with 47 cases of anaphylaxis. Brighton and Symposium definitions had sensitivities of 0.681 and 0.671, respectively, and specificities of 0.790 and 0.704, respectively. A discordant result was found in 36/128 cases (28.1%; kappa = 0.414 [95% CI 0.253, 0.574]), which represents a moderate level of agreement between case definitions. The Brighton case definition has a similar diagnostic concordance to the Symposium case definition. It does not seem to over- or underestimate cases and is sufficiently unique that the identification of an allergic trigger does not have to form part of the case definition. This will be important in the recognition of anaphylaxis resulting from the administration of drug and vaccines, where causality should be examined separately from case ascertainment. PMID- 20000868 TI - Association between drug and vaccine use and acute immune thrombocytopenia in childhood: a case-control study in Italy. AB - Immune thrombocytopenic purpura (ITP) is an immunomediated disease characterized by a decrease in platelet count and, in its more severe forms, by bleeding symptoms. Many drugs have been implicated in the pathogenesis of drug-induced thrombocytopenia in adults; only limited data on drug-related ITP in children have been published. Our study was set up to evaluate the consistency of the association between drug and vaccine use and ITP in children. This study is part of an Italian multicentre study on adverse drug reactions in children, coordinated by the Italian National Institute of Health, which was started in November 1999 and is ongoing. The study was conducted by enrolling all children aged more than 1 month who were hospitalized through the paediatric emergency department for the following conditions: thrombocytopenia (platelet count <100 x 103/L); acute neurological disorders; non-infectious mucocutaneous diseases and vasculitis; and endoscopically confirmed gastroduodenal lesions and/or clinically defined haematemesis and melaena. Children with chronic pathologies or concomitant diagnoses of cancer or immunodeficiency were not included in our study. During hospital admission, a physician interviewed parents using a structured questionnaire. The main aim of the interview was to collect information on drug exposure in a time period of 3 weeks and vaccine exposure in a period of 6 weeks preceding hospitalization. Using a case-control study design, exposure of children with thrombocytopenia (cases) to drugs and vaccines was compared with similar exposure of children with gastroduodenal lesions and neurological disorders (controls); this allowed us to estimate the odds ratios (ORs) of the occurrence of thrombocytopenia associated with the use of drugs or vaccines. Up to December 2007, the study population included 387 cases of thrombocytopenia and 1924 controls. Despite the low platelet count, ITP was generally a mild disease, without serious bleeding in the majority of cases and associated with a short length of hospital stay. After adjusting for concurrent use of other drugs, use of the antibacterials was associated with a more than 2 fold increase in the risk of developing ITP (OR 2.4; 95% CI 1.8, 3.1). Mucolytics and NSAIDs were associated with an OR of 1.9; 95% CI 1.2, 2.9 and 1.5; 95% CI 1.0, 2.1 respectively, while paracetamol (acetaminophen) was associated with an OR of 1.5; 95% CI 1.2, 2.0. MMR vaccination was associated with an increased risk of developing ITP (OR 2.4; 95% CI 1.2, 4.7). The results of this study provide evidence for an association between ITP and exposure to selected antibacterials, NSAIDs, paracetamol, mucolytics and MMR vaccination. PMID- 20000869 TI - Long-term developmental outcome of children of women with epilepsy, unexposed or exposed prenatally to antiepileptic drugs: a meta-analysis of cohort studies. AB - Results of studies investigating the long-term effects of intra-uterine exposure to antiepileptic drugs (AEDs) on cognitive functioning are limited and conflicting. To estimate intellectual development of children prenatally exposed or unexposed to AEDs by assessing IQ scores in a systematic review and meta analysis. A literature search using Pubmed, EMBASE and Google Scholar from inception to 30 April 2009 was performed to identify all original cohort studies that investigated cognitive functioning after in utero exposure to AEDs. Studies had to include at least one group exposed to an AED and one unexposed group. Data from drug exposed and unexposed controls were combined using a random effects model. Eleven studies met the inclusion criteria. Eight studies (three for valproic acid and five for carbamazepine) evaluated IQ as a measure of cognitive development. IQ was assessed by the Wechsler, Bayley or McCarthy intelligence scales, depending on age. One study investigated phenytoin and one study investigated phenobarbital (phenobarbitone). Because one study was reported in two different publications, seven studies were included in the meta-analysis. In total, the seven selected studies included 67 children exposed in utero to valproic acid and 151 exposed to carbamazepine, and 494 unexposed controls born to healthy women or to women with untreated epilepsy. The mean full-scale IQ (FSIQ), verbal IQ (VIQ) and performance IQ (PIQ) scores in children exposed to valproic acid in utero were 83.9 (95% CI 64.2, 103.6), 93.7 (95% CI 72.6, 114.7) and 88.3 (95% CI 69.9, 106.9), respectively. The mean FSIQ, VIQ and PIQ scores in the control group were 102 (95% CI 90, 116), 101 (95% CI 87, 114) and 99 (95% CI 90, 117), respectively. The mean FSIQ, VIQ and PIQ were all significantly lower in the valproic acid group compared with the unexposed group. The FSIQ and VIQ of children exposed to carbamazepine were not statistically different from those of the unexposed control group. In a sub-analysis of carbamazepine exposure in three studies using the Wechsler intelligence scale, PIQ was significantly lower in children exposed to carbamazepine than in unexposed children. Although our analysis revealed no evidence that untreated maternal epilepsy was associated with a lower IQ in the child, there may have been confounding factors, such as milder epilepsy, in this group. Exposure to valproic acid in pregnancy is associated with significantly reduced intelligence in children whose mothers were treated for epilepsy. Exposure to carbamazepine in pregnancy does not appear to be associated with reduced FSIQ and VIQ in children, although PIQ was lower in the sub-analysis. Clinicians should inform families of the potential cognitive adverse effects of valproic acid. More studies are needed to corroborate these findings. PMID- 20000870 TI - The role of toll-like receptors in host defenses and their relevance to dermatologic diseases. AB - The family of toll-like receptors (TLRs) plays a central role in the cutaneous immune defense system. To date, different TLRs have been found on several major cell populations of the skin, such as keratinocytes, fibroblasts, antigen presenting cells, and melanocytes. Activation of TLRs leads, via different intracellular signaling pathways, to the production of pro-inflammatory stimuli, and is considered a danger signal that should transform the skin in to the functional state of defense. However, TLRs have also been implicated in tissue homeostasis and renewal. Within the group of TLRs, two types have been identified: surface-expressed TLRs, which are predominantly active against bacterial cell wall compounds; and intracellular receptors, which preferentially recognize virus-associated pattern molecules. In addition, surface-expressed receptors trigger phagocytotic and maturation signals, while the intracellular TLRs lead to the induction of antiviral genes. Our review aims to outline the importance of TLRs in the pathogenesis of numerous skin diseases and the potential of TLR agonists as a treatment option for various skin diseases. PMID- 20000871 TI - Complementary and alternative medicine in alopecia areata. AB - Alopecia areata is an unpredictable hair-loss condition. As there is no cure for alopecia areata and no effective conventional therapy, a substantial number of alopecia areata patients resort to complementary and alternative medical remedies and therapies (CAM). This review on the application of CAM in alopecia areata addresses two pertinent aspects. First, it provides a current overview of the published medical literature on CAM used in alopecia areata, and alopecia areata related studies. Second, it presents a thorough assessment of the considerations and limitations of the use of CAM for the treatment of alopecia areata. A systematic MEDLINE search yielded 13 studies of the clinical use of CAM in the management of alopecia areata, all belonging to one of the five main categories of CAM. Methodological quality was analyzed using objective assessment scores (Wilson and Lawrence scores). Unfortunately, no study was of sufficient internal validity to provide robust evidence of the benefit of CAM. This might be attributable to several specific disease characteristics of alopecia areata, which require an especially solid trial design to properly assess the therapeutic effects of CAM. The review concludes with some recommendations for improving the quality of trials incorporating CAM in the treatment of alopecia areata. PMID- 20000872 TI - A current review of juvenile pemphigus vulgaris: analysis of data on clinical outcomes. AB - Forty-seven cases of juvenile pemphigus vulgaris have been reported in the English literature. Histology of lesional skin and direct immunofluorescence of perilesional skin are both necessary for a complete diagnosis. The autoimmune bullous condition can affect the skin and mucous membranes individually, but typically affects both concurrently. Disease characteristics in juvenile patients are similar to those in adults; however, a disruption of biologic and social development is of particular concern during adolescence. Although systemic corticosteroids have been used to successfully treat the disease in most cases, long-term use is often necessary for adequate control. Adverse effects from therapy can have devastating effects during this critical period of hormonal changes, physical and mental growth, and social and cultural development that occurs during adolescence. Newer therapies must be designed to adequately treat juvenile patients while also limiting serious adverse effects. PMID- 20000873 TI - Antibacterial prophylaxis in dermatologic surgery: an evidence-based review. AB - Clean, non-contaminated skin surgery is associated with low rates of surgical site infection (SSI), bacterial endocarditis, and joint prosthesis infection. Hence, antibacterial prophylaxis, which may be associated with adverse effects, the emergence of multidrug-resistant pathogens, and anaphylaxis, is generally not recommended in dermatologic surgery. Some body sites and surgical reconstructive procedures are associated with higher infection rates, and guidelines for SSI antibacterial prophylaxis have been proposed for these cases. Large prospective, controlled trials are needed to ascertain the role of oral SSI prophylaxis for these surgical sites and procedures especially in patients with diabetes mellitus who are intrinsically at greater risk of SSI. Topical antibacterial ointment and sterile paraffin appear to make no difference to healing or the incidence of SSIs in clean wounds. Although further research is needed, preliminary studies have shown that intraincisional antibacterials, which may be associated with fewer adverse effects and a lower risk of multidrug-resistant bacteria, could potentially be helpful for SSI prophylaxis. Trials using honey- and silver impregnated dressings have found no advantage in the healing of chronic wounds. However, several case studies, which need corroboration in larger studies, suggest that these dressings may be helpful in preventing and treating SSIs. Bacterial endocarditis and joint prosthesis infection prophylaxis are not routinely recommended in cutaneous surgery. The updated 2007 American Heart Association guidelines now advocate bacterial endocarditis prophylaxis for high risk cardiac patients having surgery involving the oral mucosa or infected skin. The latest American Dental Association/American Academy of Orthopaedic Surgery guidelines recommend considering antibacterial prophylaxis for oral procedures where bleeding is anticipated and for surgery involving acute orofacial skin infections if the patient has had a total joint replacement within 2 years or is in a high-risk group and has had a joint replacement at any time. PMID- 20000874 TI - Topical treatment of actinic keratoses with piroxicam 1% gel: a preliminary open label study utilizing a new clinical score. AB - BACKGROUND: The cyclo-oxygenase enzymes 1 and 2 (COX-1 and COX-2) are both involved in skin tumorigenesis, causing inhibition of apoptosis, angiogenesis, invasiveness, recruitment of growth factors, immunosuppression, and production of carcinogens. Piroxicam is a nonselective nonsteroidal anti-inflammatory drug that blocks the activity of COX-1 and COX-2. OBJECTIVE: To evaluate the efficacy and tolerability of piroxicam 1% gel in the treatment of actinic keratoses. METHODS: Piroxicam 1% gel was applied twice daily for 12 weeks to 31 actinic keratoses. The lesions were evaluated clinically and by means of dermoscopy at an initial baseline visit, at intermediate visits, and after 90 days. Changes were evaluated using a new scoring system (AKESA), based on the clinical presence of erythema, scale, and atrophy on a target lesion. In our experience, the use of piroxicam 1% gel for 90 days induced complete regression in 48% of evaluated actinic keratoses, corresponding to keratotic and verrucous clinical variants. In these lesions, the AKESA score was markedly reduced after treatment. Adverse effects were pruritus, mild erythema, dry skin, and, rarely, rash. Our preliminary trial shows that piroxicam exerts anti-tumorigenic effects and may play a useful role in the chemoprevention of skin cancers. PMID- 20000875 TI - Topical pimecrolimus 1% cream for resistant seborrheic dermatitis of the face: an open-label study. AB - BACKGROUND: Treatment options for seborrheic dermatitis are numerous, including both topical and systemic agents (e.g. topical corticosteroids, oral antifungals, and psoralen plus UVA). However, long-term use of topical corticosteroids may lead to adverse effects. Pimecrolimus 1% cream is an effective and well tolerated treatment for seborrheic dermatitis. OBJECTIVE: To explore the efficacy of pimecrolimus 1% cream for the treatment of seborrheic dermatitis lesions resistant to conventional treatments. METHODS: Sixteen patients with resistant seborrheic dermatitis of the face applied pimecrolimus 1% cream twice daily for 2 weeks. The lesions were assessed clinically and the severity of the signs were assessed using a 4-point score. Additionally, the scores of all affected regions (paranasal, forehead, and eyebrows) were evaluated separately to assess whether different results would be obtained in different regions of the face. Also, patients completed self-assessments on a 100 mm Visual Analogue Scale (VAS) at each visit. RESULTS: Statistically significant reductions in the scores of all parameters were observed at day 7 and day 14 of the study. There were no significant differences between the responses on the three regions of the face. No adverse effects were reported except for temporary pruritus immediately after the application of pimecrolimus 1% cream in one patient. CONCLUSION: Although the interpretation of efficacy was limited by the open-label, non-controlled study design and the small number of patients, this trial suggests that pimecrolimus 1% cream may be a successful treatment choice for patients with resistant seborrheic dermatitis of the face. PMID- 20000876 TI - Trends in dermatology publications over the past 15 years. AB - BACKGROUND: There is a continually increasing amount of medical literature, which makes the challenge of keeping up to date in the field of dermatology increasingly difficult. OBJECTIVE: To test the hypothesis that the total number and the number in various categories of publications in pediatric and adult dermatology have increased over time in a linear fashion. METHODS: We evaluated all MEDLINE articles from 1 January 1993 to 31 December 2007. We limited the search to 'skin' AND 'diseases' OR 'dermatology' for adults and children. We used regression analysis to determine the effect of the year of publication on the number of publications of each type. RESULTS: MEDLINE reported 17, 925 publications in adult dermatology and 9011 publications in pediatric dermatology during the evaluation period. There was a significant linear increase in the number of publications over the study period in both categories. There was a steady and similar increase over time in both pediatric and adult dermatology in total publications, randomized controlled trials, clinical trials, case reports, and letters to the editor, while there were too few meta-analyses, editorials, and clinical guidelines to make meaningful analyses of trends. CONCLUSIONS: The fields of pediatric and adult dermatology have had a significant yearly increase in studies providing a high level of scientific evidence with a much slower rise in the number of articles providing a lower level of evidence. PMID- 20000877 TI - Coexistence of basal cell carcinomas and multiple sebaceous gland hyperplasias in a cyclosporine (ciclosporin)-treated renal transplant recipient. AB - A 55-year-old man presented with multiple, asymptomatic, yellowish papules on his face with a 4-year history, and two non-healing tumoral lesions on his nose with a 7-month history. He was a renal transplant recipient and had been treated with cyclosporine (ciclosporin) for 9 years. A biopsy from the asymptomatic, yellowish papule on the face showed sebaceous gland hyperplasia, and biopsies from the lesions on the nose revealed basal cell carcinomas. The lesions on the nose were excised. Sebaceous gland hyperplasia and skin cancers are among the cutaneous neoplasms observed in renal transplant recipients receiving cyclosporine. To our knowledge, this is the third reported case of the coexistence of basal cell carcinomas and multiple sebaceous gland hyperplasias in a cyclosporine-treated renal transplant recipient. PMID- 20000878 TI - Cutaneous collagenous vasculopathy: ultrastructural and immunohistochemical study of a new case. AB - Cutaneous collagenous vasculopathy is a very rare entity first described in 2000, manifesting clinically with acquired, progressively diffuse, cutaneous telangiectases with a histologically distinct aspect, characterized by a thick hyaline collagenous wall. Its cause is unknown. Differential diagnosis includes mainly generalized essential telangiectasia and telangiectatic mastocytosis. We report a 65-year-old Caucasian patient who presented with progressive telangiectasia starting over the lower limbs that spread progressively upwards. Histopathologic (light and electron microscopic) examinations were typical of cutaneous collagenous vasculopathy. We review the salient clinicopathologic features of this poorly known condition. PMID- 20000879 TI - Cutaneous xanthogranulomas, hepatosplenomegaly, anemia, and thrombocytopenia as presenting signs of juvenile myelomonocytic leukemia. AB - The development of xanthogranulomas has been linked to hematologic malignancies in children and adults, based on a number of reports in the literature. In children, a specific association between juvenile xanthogranuloma, neurofibromatosis 1, and juvenile myelomonocytic leukemia has been described. We report a case of a 9-month-old child, without a known diagnosis of neurofibromatosis 1, who presented with hepatosplenomegaly, anemia, thrombocytopenia, and multiple cutaneous nodules, which were confirmed to be juvenile xanthogranulomas upon biopsy. A concurrent work-up showed that the child had juvenile myelomonocytic leukemia. Although cutaneous juvenile xanthogranulomas are benign lesions, in several reported cases they have been shown to herald leukemia. This association between xanthogranulomas and hematologic malignancy is poorly understood. Juvenile xanthogranulomas have a number of morphologic variants and clinical presentations that can be confused with the cutaneous lesions of Langerhans cell histiocytosis and dermatofibroma. Recognition of the broad clinicopathologic spectrum of juvenile xanthogranulomas is critical for proper diagnosis. PMID- 20000880 TI - Recurrent episodes of painful and pruritic red skin lesions. PMID- 20000881 TI - Markers of inflammation and cardiovascular disease: clinical applications of C reactive protein determination. AB - C-reactive protein (CRP) is produced by the macrophages in the liver and adipocytes and is integrated in the acute-phase response pathway. Being a nonspecific marker of inflammation, it increases in response to inflammation. The results of recent studies that have analyzed the role of CRP have not yet influenced current clinical practice. When used in combination with other established biomarkers for the prediction of the first major cardiovascular event or death, CRP does not improve the risk stratification obtained with the current guidelines. The reduction of CRP levels itself or as a statin-related pleiotropic effect has been assessed in different scenarios, including the acute phase of myocardial infarction; secondary prevention of cardiovascular diseases; special patient populations, such as diabetic patients; and finally in a primary prevention study (JUPITER [Justification for the Use of statins in primary Prevention: an Intervention Trial Evaluating Rosuvastatin]). Risk stratification in all the examined scenarios was related to serum LDL-C levels; in other words, the degree of cardiovascular risk was always lipid dependent. PMID- 20000882 TI - Role of heart rate in cardiovascular diseases: how the results of the BEAUTIFUL study change clinical practice. AB - The BEAUTIFUL (morBidity-mortality EvAlUaTion of the If inhibitor ivabradine in patients with coronary artery disease and left ventricULar systolic dysfunction) study assessed the morbidity and mortality benefits of the HR-lowering agent ivabradine. The placebo arm of the BEAUTIFUL trial was a large cohort of patients with stable coronary artery disease (CAD) and left ventricular systolic dysfunction. A subanalysis in the placebo group tested the hypothesis that elevated resting HR at baseline was a marker for subsequent cardiovascular death and morbidity. The primary aim of the study was to test whether lowering the HR with ivabradine reduced cardiovascular death and morbidity in patients with CAD and left ventricular systolic dysfunction. In the overall analysis, reduction in HR with ivabradine did not improve cardiac outcomes compared with placebo. The most important finding of the study was that patients with high baseline HR had an increase in serious cardiovascular events including death (34%), hospital admission secondary to congestive heart failure (53%), acute myocardial infarction (46%), or revascularization procedure (38%). In addition, in the subset analysis focusing on patients with baseline HR > or =70 bpm and left ventricular ejection fraction <40% the agent resulted in a 36% decrease in hospital admissions secondary to fatal and nonfatal myocardial infarction and a 30% decrease in coronary revascularization. The first practical implication from the study includes that baseline HR should be recorded in addition to other risk factors such as BP and lipid profile, in the follow-up of patients with CAD. Attempts should be made to achieve HR <70 bpm by cardiac rehabilitation and routine use of appropriately dosed beta-blockers. Despite the neutral results obtained in the BEAUTIFUL study, ivabradine could be administered to the subgroup of patients in whom HR <70 bpm is not achieved despite proper dosing of beta blockers and in those in whom beta-blockers are contraindicated. Furthermore, in clinical practice, ivabradine may be helpful for patients with stable CAD who have a high HR while receiving beta-blockers. Future studies are needed to confirm the hypothesis that single reduction of HR can improve cardiovascular prognosis. PMID- 20000883 TI - Update on antiplatelet therapy in acute coronary syndromes: what do new drugs bring into clinical practice? AB - The incidence of new coronary events in patients receiving dual antiplatelet therapy (e.g. cyclo-oxygenase inhibitors such as aspirin [acetylsalicylic acid; ASA]) and ADP receptor blockers (e.g. clopidogrel) is high. Therefore, it is critical to identify patients who require more intense treatment such as those with poor tolerance to existing drugs, those with genotypes that predict treatment resistance, diabetic patients, and smokers. The new ADP receptor blockers (prasugrel, cangrelor, Ticagrelor) can provide greater efficacy but it should not be associated with increased bleeding. Thrombin receptor antagonists (e.g. SCH530348) are another alternative that is currently being tested in randomized trials. PMID- 20000884 TI - Current analysis of the results of the COURAGE trial: has an optimal treatment been reached in clinical practice? AB - The COURAGE (Clinical Outcomes Utilizing Revascularization and Aggressive Drug Evaluation) trial was designed to evaluate the benefits of percutaneous coronary intervention in reducing the risk of cardiovascular events in patients with chronic stable coronary artery disease. The results reinforce the prior evidence regarding the importance of medical treatment which should be a universal goal. The study population enrolled was quite average for a routine cardiology clinic. However, the results of cardiac intervention are center-dependent and therefore need to be analyzed as such. Patients should initially receive the optimal medical treatment. Patients with symptom persistence, intolerance to medical treatment, and moderate to severe ischemia should be considered candidates for combined treatment. Treatment needs to be individualized and discussed with the patient. New studies, without the limitations of the COURAGE trial, enrolling high-risk patients treated with new interventional technologies, are needed to assess the impact of ischemia in long-term prognosis. PMID- 20000885 TI - Is it time to discard beta-blockers in patients undergoing noncardiac surgery? AB - Prevention of cardiac complications in patients undergoing major surgical or vascular procedures should be approached as the prevention of these complications in other common situations. beta-Adrenoceptor antagonists (beta-blockers) should be indicated in patients who are candidates for those treatments, regardless of whether surgery is performed. Current guidelines include patients with acute coronary syndrome, systolic dysfunction, chronic coronary artery disease (CAD), vascular disease, and diabetes as candidates for long-term beta-blocker therapy. Therefore risk stratification prior to surgery is indeed required to identify patients with existing CAD or myocardial ischemia in order to be able to treat them as such. In addition, beta-blockers may reduce ischemic complications in high-risk patients. However, initiating treatment at high dosages immediately before surgery has been associated with an increased risk of hypotensive strokes. PMID- 20000886 TI - Augmented renal clearance: implications for antibacterial dosing in the critically ill. AB - The prescription of pharmaceuticals in the critically ill is complicated by a paucity of knowledge concerning the pharmacokinetic implications of the underlying disease state. Changes in organ function can be dramatic in this population, both as a consequence of the primary pathophysiology and in response to clinical interventions provided. Vascular tone, fluid status, cardiac output and major organ blood flow can be significantly altered from baseline, influencing the volume of distribution and clearance of many commonly prescribed agents. Although measurable endpoints can be used to titrate doses for many drugs in this setting (such as sedatives), for those agents with silent pharmacodynamic indices, enhanced excretory organ function can result in unexpectedly low plasma concentrations, leading to treatment failure. This is particularly relevant to the use of antibacterials in the critically ill, where inadequate, inappropriate and/or delayed prescription can have significant effects on morbidity and mortality. Augmented renal clearance (ARC) refers to enhanced renal elimination of circulating solute and is being described with increasing regularity in the critically ill. However, defining this process in terms of current measures of renal function is problematic, as although the glomerular filtration rate (GFR) is largely considered the best index of renal function, there is no consensus on an upper limit of normal. In addition, the most readily available and accurate estimate of the GFR at the bedside is still widely debated. From a pharmacokinetic point of view, ARC can result in elevated renal elimination and subtherapeutic plasma concentrations of pharmaceuticals, although whether this process solely involves augmented filtration (as opposed to enhanced tubular secretion and/or reabsorption) remains uncertain. The primary contributors to this process are likely to be the innate immune response to infection and inflammation (with its associated systemic and haemodynamic consequences), fluid loading and use of vasoactive medications. The resultant increase in cardiac output and renal blood flow prompts enhanced glomerular filtration and drug elimination. Current evidence suggests that young patients without pre-existing co-morbidity or organ dysfunction who present with trauma are most likely to manifest ARC. As this phenomenon has received little attention in the literature, dose modification has rarely been considered. However, with increasing data supporting the concept, and many investigators demonstrating subtherapeutic concentrations of drugs in the critically ill, consideration of ARC and alternative dosing regimens is now mandatory, both to improve the likelihood of treatment success and to reduce the rate of development of antibacterial resistance. PMID- 20000888 TI - Absorption and excretion of colestilan in healthy subjects. AB - Colestilan, an anion-exchange resin binding both phosphate and bile-acid anions, is under development for the treatment of hyperphosphataemia and dyslipidaemia, which occur in the majority of end-stage renal disease patients. This study using 14C-colestilan was conducted to investigate the absorption and excretion of colestilan in humans. Following a 28-day run-in period with administration of colestilan 3 g three times daily, 12 subjects received a single oral dose of 14C colestilan 100 mg (approximately 4.0 MBq) and colestilan 3 g under fasted conditions on the morning of day 1. A total of 9 g of colestilan was administered three times daily on days 1-4. Total radioactivity levels in whole blood (at 4, 8, 12 and 24 hours and then at 24-hour intervals) and in the urine and faeces (from 0 to 24 hours and then at 24-hour intervals) were monitored up to 216 hours postdose (day 10). Total radioactivity measured in all whole-blood samples was below the lower limit of quantification (0.025 microg equivalent of 14C-labelled colestilan/mL of whole blood). Total radioactivity assessed in all urine samples was also below the lower limit of quantification (0.003 microg equivalent/mL for urine), except at 0-24 hours postdose, when 0.01% of the radioactive dose was excreted by all subjects. This level was below the predetermined water soluble impurity level of 0.04%. The mean cumulative excretion of total radioactivity in the faeces was 99.66% by 216 hours postdose, excluding one subject with incomplete collection of faecal samples. These results demonstrate that colestilan is not absorbed from the gastrointestinal tract and is completely excreted in the faeces. PMID- 20000887 TI - Intracellular Pharmacokinetics of Antiretroviral Drugs in HIV-Infected Patients, and their Correlation with Drug Action. AB - In patients infected by HIV, the efficacy of highly active antiretroviral (ARV) therapy through the blockade of different steps of the retrovirus life cycle is now well established. As HIV is a retrovirus that replicates within the cells of the immune system, intracellular drug concentrations are important to determine ARV drug efficacy and toxicity. Indeed, nucleoside reverse transcriptase inhibitors (NRTIs), non-NRTIs (NNRTIs), newly available integrase inhibitors and protease inhibitors (PIs) act on intracellular targets. NRTIs are prodrugs that require intracellular anabolic phosphorylation to be converted into their active form of triphosphorylated NRTI metabolites, most of which have longer plasma half lives than their parent compounds. The activity of intracellular kinases and the expression of uptake transporters, which may depend on cell functionality or their activation state, may greatly influence intracellular concentrations of triphosphorylated NRTI metabolites. In contrast, NNRTIs and PIs are not prodrugs, and they exert their activity by inhibiting enzyme targets directly. All PIs are substrates of cytochrome P450 3A, which explains why most of them display poor pharmacokinetic properties with intensive presystemic first-pass metabolism and short elimination half-lives. There is evidence that intracellular concentrations of PIs depend on P-glycoprotein and/or the activity of other efflux transporters, which is modulated by genetic polymorphism and coadministration of drugs with inhibiting or inducing properties. Adequate assay of the intracellular concentrations of ARVs is still a major technical challenge, together with the isolation and counting of peripheral blood mononuclear cells (PBMCs). Furthermore, intracellular drug could be bound to cell membranes or proteins; the amount of intracellular ARV available for ARV effectiveness is never measured, which is a limitation of all published studies. In this review, we summarize the findings of 31 studies that provided results of intracellular concentrations of ARVs in HIV-infected patients. Most studies also measured plasma concentrations, but few of them studied the relationship between plasma and intracellular concentrations. For NRTIs, most studies could not establish a significant relationship between plasma and triphosphate concentrations. Only eight published studies reported an analysis of the relationships between intracellular concentrations and the virological or immunological efficacy of ARVs in HIV patients. In prospective studies that were well designed and had a reasonable number of patients, virological efficacy was found to correlate significantly with intracellular concentrations of NRTIs but not with plasma concentrations. For PIs, the only prospectively designed trial of lopinavir found that virological efficacy was influenced by both trough plasma concentrations and intracellular concentrations. ARVs are known to cause important adverse effects through interference with cellular endogenous processes. The relationship between intracellular concentrations of ARVs and their related toxicity was investigated in only four studies. For zidovudine, the relative strength of the association between a decrease in haemoglobin levels and plasma zidovudine concentrations, as compared with intracellular zidovudine triphosphate concentrations, is still unknown. Similarly, for efavirenz and neuropsychological disorders, methodological differences confound the comparison between studies. In conclusion, intracellular concentrations of ARVs play a major role in their efficacy and toxicity, and are influenced by numerous factors. However, the number of published clinical studies in this area is limited; most studies have been small and not always adequately designed. In addition, standardization of assays and PBMC counts are warranted. Larger and prospectively designed clinical studies are needed to further investigate the links between intracellular concentrations of ARVs and clinical endpoints. PMID- 20000890 TI - Exploiting forward scattering for detecting submerged proud/half-buried unexploded ordnance. AB - Laboratory underwater bistatic scattering measurements are reported for free, proud, and half-buried unexploded ordnances for 0 degrees and 90 degrees source angles. Forward echoes are larger than backscattered returns, and half burial significantly decreases the latter but not the former. Results agree with analytic predictions borrowed from radar. The forward echo and source signal are separated by measurements made with and without the target, a method not possible in a target search. For this, a method is described that uses knowledge of the source location and the hyperbolic character in time-cross range of the signals received at points along a line. PMID- 20000889 TI - Semi-mechanistic population pharmacokinetic drug-drug interaction modelling of a long half-life substrate and itraconazole. AB - BACKGROUND: For compounds with a long elimination half-life, the evaluation of a drug-drug interaction (DDI) study can be challenging. The standard analytical approach of a non-compartmental analysis (NCA) might not be able to detect the full interaction potential and may lead to a significant underestimation of the interaction. The most appropriate method for data analysis might be a semi mechanistic population pharmacokinetic modelling approach. OBJECTIVES: To accomplish a semi-mechanistic DDI model for a long-elimination-half-life drug substrate, tesofensine, and the cytochrome P450 (CYP) 3A4 inhibitor itraconazole, and to compare the results of the semi-mechanistic model with the results obtained from the standard NCA approach. Additionally, the impact of different schedules of itraconazole on tesofensine pharmacokinetics and the general performance of the standard NCA approach were evaluated. METHODS: Overall, 28 subjects received a single oral dose of tesofensine 2 mg; 14 of these subjects were coadministered an oral itraconazole 400 mg loading dose and a 200 mg maintenance dose for 6 days before and 5 days after administration of tesofensine. The dataset contained 465 plasma concentrations of tesofensine (full profiles) and 80 plasma concentrations of itraconazole (trough values). First, pharmacokinetic models of itraconazole and tesofensine were developed in parallel. Subsequently, a combined model was developed, taking into account CYP3A4 inhibition. The analyses were performed using NONMEM software. RESULTS: The plasma concentration-time profiles of itraconazole and tesofensine were best described by a one-compartment model for each drug, with first-order elimination rate constants that were both inhibited by itraconazole concentrations. Inhibition resulted in reduced clearances and prolonged elimination half-lives for tesofensine and itraconazole: using NCA, the actual study revealed an approximately 9% increase in exposure for the timeframe of the coadministration with itraconazole (the area under the plasma concentration-time curve (AUC) from 0 to 144 hours [AUC(144h)]), and the impact on exposure estimated to infinity (AUC(infinity)) was approximately 26%. These results are in contrast to the model predicted results, where the inhibitory effect of itraconazole caused a 38% reduction in the clearance of tesofensine, leading to a 63% increased exposure. CONCLUSIONS: This analysis presents a semi-mechanistic population pharmacokinetic approach that may be useful for the evaluation of DDI studies. The model can be an aid in evaluating DDI studies for compounds with a long elimination half-life, especially when the inhibitor cannot be administered over a sufficient period. Additionally, the population model-based approach may allow simplification of the design and the analysis and interpretation of safety and efficacy findings in DDI studies. PMID- 20000891 TI - Masking with interaurally "double-delayed" stimuli: the range of internal delays in the human brain. AB - Is binaural processing in humans different to that of other mammals? While psychophysical data suggest that the range of internal delays necessary for processing interaural time differences is at least +/-3 ms, physiological data from small mammals indicate a more limited range. This study demonstrates that binaural detection is impeded by reduced interaural coherence in auditory channels remote from the signal frequency, in accordance with the wider critical bandwidths reported for binaural processing. This explains previous psychophysical data without requiring long internal delays. The current psychophysical data support the view that human binaural processing is similar to that of other mammals. PMID- 20000892 TI - Accuracy of the deterministic travel time retrieval from cross-correlations of non-diffuse ambient noise. AB - Measurements of long-range cross-correlations of ambient noise underlie acoustic noise interferometry, a promising technique for passive remote sensing of the environment. Previously established simple, exact relations between deterministic Green's functions and the cross-correlation function of perfectly diffuse noise do not necessarily hold for noise fields in the ocean and atmosphere. Here, the method of a stationary phase is applied to study the information content of the cross-correlation function of non-diffuse noise and to quantify the accuracy of passive measurements of the acoustic travel times. PMID- 20000893 TI - Acoustic measurements of clay-size particles. AB - Knowledge of sediment concentration is important in the study of streams and rivers. The work presented explores using high frequency (20 MHz) acoustic signal attenuation to measure the concentration of fine sediment particles (0.2-5.0 microm) in a fluvial environment. A small laboratory tank with a pitch-catch transducer configuration measured a 35 dB change in signal level over a wide range of kaolinite and bentonite concentrations (1-14 g/l) over a range of distances (180-357 mm). The data suggest that a fixed distance of 180 mm between the transducers will be capable of measuring the entire range of concentrations. PMID- 20000894 TI - The intelligibility of pointillistic speech. AB - A form of processed speech is described that is highly discriminable in a closed set identification format. The processing renders speech into a set of sinusoidal pulses played synchronously across frequency. The processing and results from several experiments are described. The number and width of frequency analysis channels and tone-pulse duration were variables. In one condition, various proportions of the tones were randomly removed. The processed speech was remarkably resilient to these manipulations. This type of speech may be useful for examining multitalker listening situations in which a high degree of stimulus control is required. PMID- 20000895 TI - A special relation between Young's modulus, Rayleigh-wave velocity, and Poisson's ratio. AB - Bayon et al. [(2005). J. Acoust. Soc. Am. 117, 3469-3477] described a method for the determination of Young's modulus by measuring the Rayleigh-wave velocity and the ellipticity of Rayleigh waves, and found a peculiar almost linear relation between a non-dimensional quantity connecting Young's modulus, Rayleigh-wave velocity and density, and Poisson's ratio. The analytical reason for this special behavior remained unclear. It is demonstrated here that this behavior is a simple consequence of the mathematical form of the Rayleigh-wave velocity as a function of Poisson's ratio. The consequences for auxetic materials (those materials for which Poisson's ratio is negative) are discussed, as well as the determination of the shear and bulk moduli. PMID- 20000896 TI - Authors' Response to Strasberg's "Comment on 'Measurement of the frequency dependence of the ultrasonic parametric threshold amplitude for a fluid-filled cavity' " [J. Acoust. Soc. Am. 125, 1857 (2009)]. AB - This letter is a response to Strasberg's recent paper, "Comment on 'Measurement of the frequency dependence of the ultrasonic parametric threshold amplitude for a fluid-filled cavity.'" The authors dispute the conclusions of Strasberg regarding the effect observed by Teklu et al. [J. Acoust. Soc. Am. 120, 657-660 (2006)] published previously in JASA. PMID- 20000897 TI - Broadband directive sources for acoustic discrete-time simulations. AB - This work investigates a generalized implementation of directive sources on discrete-time methods. Results based on previous work for sinusoidal sources have been extended to accomplish real life source requirements, which have a frequency dependent pattern and show gradual transitions between the different directivities at successive frequencies. One of the main advantages of this method lies in its capacity to be applied to any discrete-time method, since it is based on a proper combination of impulse responses radiated from discrete positions, which, in turn, will produce a particular pressure distribution at far field. Furthermore, some considerations about mesh-dispersion effects on source modeling are addressed using some examples. PMID- 20000899 TI - On the use of leaky modes in open waveguides for the sound propagation modeling in street canyons. AB - An urban, U-shaped, street canyon being considered as an open waveguide in which the sound may propagate, one is interested in a multimodal approach to describe the sound propagation within. The key point in such a multimodal formalism is the choice of the basis of local transversal modes on which the acoustic field is decomposed. For a classical waveguide, with a simple and bounded cross-section, a complete orthogonal basis can be analytically obtained. The case of an open waveguide is more difficult, since no such a basis can be exhibited. However, an open resonator, as displays, for example, the U-shaped cross-section of a street, presents resonant modes with complex eigenfrequencies, owing to radiative losses. This work first presents how to numerically obtain these modes. Results of the transverse problem are also compared with solutions obtained by the finite element method with perfectly mathed layers. Then, examples are treated to show how these leaky modes can be used as a basis for the modal decomposition of the sound field in a street canyon. PMID- 20000898 TI - Effects of voicing in the recognition of concurrent syllables. AB - This letter reports a study designed to measure the benefits of voicing in the recognition of concurrent syllables. The target and distracter syllables were either voiced or whispered, producing four combinations of vocal contrast. Results show that listeners use voicing whenever it is present either to detect a target syllable or to reject a distracter. When the predictable effects of audibility were taken into account, limited evidence remained for the harmonic cancellation mechanism thought to make rejecting distracter syllables more effective than enhancing target syllables. PMID- 20000900 TI - Three dimensional finite element modeling of smart foam. AB - The "smart foam" concept and design originate from the combination of the passive dissipation capability of foam in the medium and high frequency ranges and the active absorption ability of piezoelectric actuator (generally polyvinylidene fluoride) in the low frequency range. This results into a passive/active absorption control device that can efficiently operate over a broad range of frequencies. In this paper, a full three dimensional finite element model of smart foam is presented including its experimental validation. The modeling tool uses quadratic poroelastic elements, as well as elastic, fluid, and piezoelectric elements. The weak integral formulation of the different media involved is presented with the associated coupling conditions. A simplified orthotropic model of poroelastic media is presented. To validate the developed model, a prototype of a smart foam has been realized and its passive absorption and radiation measured in an impedance tube and compared to predictions. The experimental validation demonstrates the validity of the model. This modeling tool constitutes a general platform to simulate and optimize various configurations of smart foams. PMID- 20000901 TI - Short-range shock formation and coalescence in numerical simulation of broadband noise propagation. AB - The number of jet and rocket noise studies has increased in recent years as researchers have sought to better understand aeroacoustic source and radiation characteristics. Although jet and rocket noise is finite-amplitude in nature, little is known about the existence of shock formation and coalescence close to the source. A numerical experiment is performed to propagate finite-amplitude noise and determine the extent of the nonlinearity over short distances with spherical spreading. The noise is filtered to have a haystack shape in the frequency domain, as is typical of such sources. The effect of the nonlinearity is compared in both the temporal and frequency domains as a function of distance. Additionally, the number of zero-crossings and overall sound pressure level is compared at several distances. The results indicate that the center frequency plays a particularly important role in the amount of coalescence and spectral redistribution that occurs. The general applicability of these results to actual near-field finite-amplitude jet and rocket noise experiments is also presented. PMID- 20000902 TI - Geostatistical modeling of sound propagation: Principles and a field application experiment. AB - The assessment of noise sources for environmental purposes requires reliable methods for mapping. Numerical models are well adapted for sophisticated simulations and sensitivity analyses; however, real-time mapping of large frequency bands must be based on fast and acceptable computations and honor in situ measurements. In this paper, a real-time mapping procedure of noise exposure is proposed. The procedure is based on geostatistical modeling of spatial variations and applied to a case study taken from an experimental campaign, where a point source was placed on a flat meadow. An analytical approximation of the acoustic field was first computed with the Embleton model. The difference between this approximation and the actual measurements (L(eq15 min) 1/3-octave bands samples from 19 microphones spread over the meadow) showed spatial structure, which has been modeled with a variogram. Finally, the geostatistical technique of kriging with external drift provided an optimal interpolation of the acoustic field data while encapsulating the first approximation from the Embleton model. Systematic geostatistical inference and real-time mapping with the proposed procedure can be envisaged in simple cases. PMID- 20000903 TI - Measurements and inversion of acoustic scattering from suspensions having broad size distributions. AB - Measurements are presented from a multi-frequency acoustic backscatter study of aqueous suspensions of irregularly shaped quartz sediments having broad particle size distributions. Using the backscattered sound from a homogenous suspension, measurements of the ensemble backscatter form function and ensemble normalized total scattering cross section were obtained. Three different size distribution types are examined; namely Gaussian, log-normal, and bi-modal distributions, each covering a range of particle sizes similar to those observed in sandy marine environments near the seabed. The measurements of ensemble scattering are compared with theoretical predictions, derived by integrating the intrinsic scattering properties of the sediments over the probability density functions of the size distributions used in the present study. The results show that the ensemble scattering parameters are significant functions of both the width and type of size distribution in suspension. The impact of errors in size distribution width on inversion predictions of both mean size and suspended concentration is also examined. The validity of the theoretical predictions is discussed, along with the implication of the inversion results for using acoustic backscatter data to measure suspended concentration and particle size in sandy marine environments. PMID- 20000904 TI - Automatic recognition of fin and blue whale calls for real-time monitoring in the St. Lawrence. AB - Monitoring blue and fin whales summering in the St. Lawrence Estuary with passive acoustics requires call recognition algorithms that can cope with the heavy shipping noise of the St. Lawrence Seaway and with multipath propagation characteristics that generate overlapping copies of the calls. In this paper, the performance of three time-frequency methods aiming at such automatic detection and classification is tested on more than 2000 calls and compared at several levels of signal-to-noise ratio using typical recordings collected in this area. For all methods, image processing techniques are used to reduce the noise in the spectrogram. The first approach consists in matching the spectrogram with binary time-frequency templates of the calls (coincidence of spectrograms). The second approach is based on the extraction of the frequency contours of the calls and their classification using dynamic time warping (DTW) and the vector quantization (VQ) algorithms. The coincidence of spectrograms was the fastest method and performed better for blue whale A and B calls. VQ detected more 20 Hz fin whale calls but with a higher false alarm rate. DTW and VQ outperformed for the more variable blue whale D calls. PMID- 20000905 TI - Eigenmodal resonances of polydisperse bubble systems on a rigid boundary. AB - This paper presents theory and experimental data on the resonance frequency of systems consisting of different-sized air bubbles attached to a rigid wall. Effects of the change in resonant frequency with bubble size and distance between the bubbles were studied. It was found that the symmetric mode resonance frequency of the bubble system decreased with increasing r=R(02)/R(01), where R(01) and R(02) are the equilibrium radii of bubbles in the system. Both the symmetric and antisymmetric modes of oscillation were detected in the experiments, with the resonant frequency of the symmetric mode dominant at small bubble separation and the frequency of the antisymmetric mode dominant when the bubbles were farther apart. A linear coupled-oscillator theoretical model was used to describe the oscillations of the bubble system, in which the method of images was used to approximate the effects of the wall. It was found that there was fair to good agreement between the predictions of the coupled-oscillator model with the experimental data. PMID- 20000906 TI - Angular and frequency spectral analysis of the ultrasonic backward beam displacement on a periodically grooved solid. AB - The ultrasonic backward beam displacement, which has been shown to occur when a bounded beam is incident upon a periodically corrugated liquid-solid interface, is studied experimentally. This effect has been previously studied on a periodic water-brass interface at one particular frequency (6 MHz) and one corresponding angle of incidence (22.5 degrees), but the question has remained whether it would also exist at other frequency and angle combinations. The knowledge of whether this phenomenon is a coincidence or whether it will occur for other frequency and angle combinations contributes to a better understanding of the interaction of ultrasound with periodic structures and diffraction effects, in particular. Potential applications exist in the study of phononic crystals and in the non destructive evaluation of materials. The present work reports results from recent experiments on the same periodically grooved brass sample that was employed in the first investigations of this phenomenon. Through the examination of frequency spectra in the form of angular and classical spectrograms, the experiments reported here show the backward beam displacement to occur for multiple angles of incidence and frequencies. Furthermore, evidence is shown as to the exact cause of the backward beam displacement, namely, a backward propagating Scholte Stoneley wave. PMID- 20000907 TI - Resonant ultrasound studies of the layered perovskite system Ca2-xSrxRuO4. AB - The elastic response of the layered perovskite system Ca(2-x)Sr(x)RuO(4) (0.2< or =x< or =2) has been studied as a function of temperature and doping concentration x using resonant ultrasound spectroscopy. The elastic constants c(11) and c(44) have been obtained for three polycrystalline samples (x=1.0, 0.5, and 0.3) and show a softening trend with increasing Ca-content. In addition, the temperature dependence of the elastic response of five single-crystals (x=2.0, 1.9, 0.5, 0.3, and 0.2) has been measured. For 2.0> or =x> or =0.5, a dramatic softening over a wide temperature range is observed upon cooling, which is attributed to the rotational instability of RuO(6) octahedra (for x=2.0 and 1.9) and the static rotation of the octahedra (for x=0.5). For the Ca-rich samples (x=0.3 and 0.2), the softening occurs in a very narrow temperature range, corresponding to the structural phase transition from high-temperature tetragonal to low-temperature orthorhombic symmetry. PMID- 20000908 TI - Estimating the shell parameters of SonoVue microbubbles using light scattering. AB - Experiments were performed to measure the dynamical response of individual SonoVue microbubbles subjected to pulsed ultrasound. Three commonly used bubble dynamic models (i.e., Hoff's, Sarkar's, and linearized Marmottant's models) were compared to determine the most appropriate model for fitting to the experimental data. The models were evaluated against published optical microscopy data. The comparison suggests that it is difficult to rank these models for lipid-shelled microbubbles undergoing small-amplitude oscillations, because under these conditions the shell parameters in these models are closely related. A linearized version of the Marmottant model was used to estimate the shell parameters (i.e., shear modulus and shear viscosity) of SonoVue microbubbles from the experimental light scattering data, as a function of ambient microbubble radius. The SonoVue microbubble shell elasticity and dilatational viscosity increase with ambient bubble radius, in agreement with previously published data. The results suggest that light scattering, used in conjunction with one of several popular bubble dynamics models, is effective at characterizing microbubble response and evaluating shell parameters. PMID- 20000909 TI - The natural frequencies of microbubble oscillation in elastic vessels. AB - A theoretical model for the dynamics of a bubble in an elastic blood vessel is applied to study numerically the effect of confinement on the free oscillations of a bubble. The vessel wall deformations are described using a lumped-parameter membrane-type model, which is coupled to the Navier-Stokes equations for the fluid motion inside the vessel. It is shown that the bubble oscillations in a finite-length vessel are characterized by a spectrum of frequencies, with distinguishable high-frequency and low-frequency modes. The frequency of the high frequency mode increases with the vessel elastic modulus and, for a thin-wall vessel, can be higher than the natural frequency of bubble oscillations in an unconfined liquid. In the limiting case of an infinitely stiff vessel wall, the frequency of the low-frequency mode approaches the well-known solution for a bubble confined in a rigid vessel. In order to interpret the results, a simple two-degree-of-freedom model is applied. The results suggest that in order to maximize deposition of acoustic energy, a bubble confined in a long elastic vessel has to be excited at frequencies higher than the natural frequency of the equivalent unconfined bubble. PMID- 20000910 TI - Post-processing of guided wave array data for high resolution pipe inspection. AB - This paper describes a method for processing data from a guided wave transducer array on a pipe. The raw data set from such an array contains the full matrix of time-domain signals from each transmitter-receiver combination. It is shown that for certain configurations of an array, the total focusing method can be applied, which allows the array to be focused at every point on a pipe in both transmission and reception. The effect of array configuration parameters on the sensitivity of the proposed method to random and coherent noise is discussed. Experimental results are presented using electromagnetic acoustic transducers for exciting and detecting the S(0) Lamb wave mode in a 12-in. diameter steel pipe at 200 kHz excitation frequency. The results show that using the imaging algorithm, a 2-mm (0.08 wavelength) diameter half-thickness hole can be detected. PMID- 20000911 TI - Analytical modeling of piezoelectric ceramic transducers based on coupled vibration analysis with application to rectangular thickness poled plates. AB - The energy method for analyzing piezoelectric ceramic transducers [B. S. Aronov, J. Acoust. Soc. Am. 117, 210-220 (2005)] is applied to the treatment of transducers with mechanical systems that can be considered as two-dimensional. Analysis is made following the general outline of the theory of coupled vibration in two degrees-of-freedom systems and its extension in calculating the resonance frequencies of elastic bodies, as suggested by Giebe and Blechschmidt [Ann. Physik, Ser. 5 18, 417-485 (1933)]. The approach to the problem is illustrated with examples of piezoceramic rectangular thickness poled plates. The resonance frequencies and effective coupling coefficients are presented as functions of the plates' aspect ratio. PMID- 20000912 TI - Estimation of vehicle speed using wayside sound pressure onset rate. AB - Transportation-related environmental noise studies, particularly those associated with rail traffic, often require long term measurements near existing rights of way. In addition to the sound produced by passing vehicles, it is also useful to know their speed. Previously, speed measurements have generally required an additional piece of instrumentation, such as a second microphone, video camera, or a radar gun--resulting in added measurement cost and complexity. The present study shows how estimations of vehicle speed can be obtained using a single wayside microphone. The method is based on the rate of pressure rise as the vehicle approaches, specifically the maximum onset rate. This paper shows how the maximum onset rate depends only on the vehicle speed, the microphone distance, and the speed of sound. In order to demonstrate the feasibility of the approach field test data are presented for train passages ranging in speed from 33.5 to 55.6 m/s, as measured by a microphone located 28.65 m from the tracks. Speed estimates derived from the onset rate were generally accurate to within 8% of the independently measured speed of the train. PMID- 20000913 TI - Determination of elastic constants of generally anisotropic inclined lamellar structure using line-focus acoustic microscopy. AB - A methodology for measuring elastic constants of different phases in materials with lamellar microstructure by line-focus acoustic microscopy is developed. The material microstructure investigated is modeled by generally anisotropic multilayers arbitrarily inclined to the sample surface on which acoustic microscopy measurements are performed. To calculate surface acoustic wave (SAW) propagation in such structures quasi-static effective elastic constants are determined and compared with calculated frequency-dependent constants. As a model material, practically important, Ti-6Al-2Sn-4Zr-2Mo alloy is selected. Time resolved line-focus acoustic microscopy experiments are performed on a Ti-6242 alpha/beta single colony (Ti-6Al-2Sn-4Zr-2Mo alloy) and on a Ti-6Al alpha-phase single crystal for which elastic constants of different phases are determined using inversion of measured SAW velocities. To validate the experimental methodology, SAW velocities in an X-cut quartz crystal are measured as a function of sample orientation angle and compared with predictions based on the known elastic moduli of quartz. PMID- 20000914 TI - Sound propagation in and low frequency noise absorption by helium-filled porous material. AB - Low-frequency noise is difficult to deal with by traditional porous material due to its inherent high acoustic impedance. This study seeks to extend the effective range of sound absorption to lower frequencies by filling a low density gas, such as helium, in the porous material. Compared with conventional air-filled absorption material, the helium-filled porous material has a much reduced characteristic impedance; hence, a good impedance matching with pure air becomes more feasible at low frequencies. The acoustic properties of a series of helium filled porous materials are investigated with a specially designed test rig. The characteristic of the sound propagation in a helium-filled porous material is established and validated experimentally. Based on the measured acoustic properties, the sound absorption performance of a helium-filled absorber (HA) of finite thickness is studied numerically as well as experimentally. For a random incidence field, the HA is found to perform much better than the air-filled absorber at low frequencies. The main advantage of HA lies in the middle range of oblique incidence angles where wave refraction in the absorber enhances sound absorption. The advantage of HA as duct lining is demonstrated both numerically and experimentally. PMID- 20000915 TI - Comparison of three measurement techniques for the normal absorption coefficient of sound absorbing materials in the free field. AB - Three different techniques for evaluating the absorption coefficient of sound absorbing materials in free field conditions are discussed. One technique measures the acoustic impedance at one point nearby a specimen, the other two techniques evaluate the impedance from the transfer function of two sound pressures and two particle velocities at two points. These are called "PU method," "PP-method," and "UU-method," respectively. An iterative algorithm to estimate the acoustic impedance of the locally reactive specimen in the spherical wave field is also applied. First, the effect of receiver positions, specimen areas, and source heights to the measured normal absorption coefficient is investigated by the boundary element method. According to these investigations, the PU-method is most stable against the effect of specimen area, and the UU method is easily affected by that effect. Closer source to the specimen distance is advantageous for the signal to noise ratio of these measurement techniques, but correction for the effect of the spherical wave field has to be applied. As a finding, the iterative algorithm works for all of three techniques. Finally, the PU-method is applied experimentally with a pressure-velocity sensor and a loudspeaker in a hemi-anechoic room. As a result, the calculated results have been verified. PMID- 20000916 TI - Transmission characteristics of a tee-junction in a rectangular duct at higher order modes. AB - Numerical and experimental studies were undertaken to characterize the noise transmission and scattering properties in higher-order modes across the tee junction of a rectangular duct used in ventilation and air-conditioning systems. To measure these properties, a formulation of a transmission matrix based on the transfer function and a two-microphone method was devised. The measurement of modal sound transmission and scattering coefficients is demonstrated for a duct element in a rectangular duct. The results of numerical simulations were verified by experiments. The results show that sound transmissions of fundamental mode and higher-order modes across the main duct are high at the eigen-frequencies of the main duct and sidebranch. Weak modal coupling of the branch-modes and the traveling wave in the main duct is observed at or very close to the eigen frequencies of the sidebranch, which shifts excitation of the higher-order branch modes at higher frequencies. A decrease in sound transmission and increase in sound scattering into higher-order modes occur with excitation of the axial branch-mode. Excitation of the longitudinal branch-mode due to branch-end reflection also results in lower sound transmission of higher-order modes across the junction along the main duct. PMID- 20000917 TI - Transmission loss of double panels filled with porogranular materials. AB - Sound transmission through hollow structures found its interest in several industrial domains such as building acoustics, automotive industry, and aeronautics. However, in practice, hollow structures are often filled with porous materials to improve acoustic properties without adding an excessive mass. Recently a lot of interest arises for granular materials of low density that can be an alternative to standard absorbing materials. This paper aims to predict vibro-acoustic behavior of double panels filled with porogranular materials by using the patch-mobility method recently published. Biot's theory is a basic tool for the description of porous material but is quite difficult to use in practice, mostly because of the solid phase characterization. The original simplified Biot's model (fluid-fluid model) for porogranular material permitting a considerable reduction in data necessary for calculation has been recently published. The aim of the present paper is to propose a model to predict sound transmission through a double panel filled with a porogranular material. The method is an extension of a previous paper to take into account the porogranular material through fluid-fluid Biot's model. After a global overview of the method, the case of a double panel filled with expanded polystyrene beads is studied and a comparison with measurements is realized. PMID- 20000918 TI - Measurement of characteristic impedance and wave number of porous material using pulse-tube and transfer-matrix methods. AB - The time domain implementation of the transfer-matrix method developed by Song and Bolton [J. Acoust. Soc. Am. 107, 1131-1154 (2000)] for measuring the characteristic impedance and wave number of porous materials is described in this paper. The so called Butterworth impulse is generated in a standing wave tube with a flat frequency response over a wide frequency range. With only two microphone measurements, the transfer matrix of porous layers can easily be determined through the calculation of complex amplitudes of incident, reflected, and transmitted pulses. The procedure has been used to measure the acoustical properties of a fiber material, and good agreement was found between measured acoustical properties and predicted results by Delany and Bazley [Appl. Acoust. 3, 105-116 (1971)] semiempirical formulas. Although the error associated with the sample-edge constraint still remains, the new method has a better frequency response as a result of the system calibration process, and the optimal inter microphone distance is no longer required compared to the frequency domain implementation. PMID- 20000919 TI - High-rate synthetic aperture communications in shallow water. AB - Time reversal communication exploits spatial diversity to achieve spatial and temporal focusing in complex ocean environments. Spatial diversity can be provided easily by a vertical array in a waveguide. Alternatively, spatial diversity can be obtained from a virtual horizontal array generated by two elements, a transmitter and a receiver, due to relative motion between them, referred to as a synthetic aperture. This paper presents coherent synthetic aperture communication results from at-sea experiments conducted in two different frequency bands: (1) 2-4 kHz and (2) 8-20 kHz. Case (1) employs binary-phase shift-keying modulation, while case (2) involves up to eight-phase shift keying modulation with a data rate of 30 kbits/s divided by the number of transmissions (diversity) to be accumulated. The receiver utilizes time reversal diversity combining followed by a single channel equalizer, with frequent channel updates to accommodate the time-varying channel due to coupling of space and time in the presence of motion. Two to five consecutive transmissions from a source moving at 4 kts over 3-6 km range in shallow water are combined successfully after Doppler compensation, confirming the feasibility of coherent synthetic aperture communications using time reversal. PMID- 20000920 TI - A source separation approach to enhancing marine mammal vocalizations. AB - A common problem in passive acoustic based marine mammal monitoring is the contamination of vocalizations by a noise source, such as a surface vessel. The conventional approach in improving the vocalization signal to noise ratio (SNR) is to suppress the unwanted noise sources by beamforming the measurements made using an array. In this paper, an alternative approach to multi-channel underwater signal enhancement is proposed. Specifically, a blind source separation algorithm that extracts the vocalization signal from two-channel noisy measurements is derived and implemented. The proposed algorithm uses a robust decorrelation criterion to separate the vocalization from background noise, and hence is suitable for low SNR measurements. To overcome the convergence limitations resulting from temporally correlated recordings, the supervised affine projection filter update rule is adapted to the unsupervised source separation framework. The proposed method is evaluated using real West Indian manatee (Trichechus manatus latirostris) vocalizations and watercraft emitted noise measurements made within a typical manatee habitat in Florida. The results suggest that the proposed algorithm can improve the detection range of a passive acoustic detector five times on average (for input SNR between -10 and 5 dB) using only two receivers. PMID- 20000921 TI - Passive cavitation imaging with ultrasound arrays. AB - A method is presented for passive imaging of cavitational acoustic emissions using an ultrasound array, with potential application in real-time monitoring of ultrasound ablation. To create such images, microbubble emissions were passively sensed by an imaging array and dynamically focused at multiple depths. In this paper, an analytic expression for a passive image is obtained by solving the Rayleigh-Sommerfield integral, under the Fresnel approximation, and passive images were simulated. A 192-element array was used to create passive images, in real time, from 520-kHz ultrasound scattered by a 1-mm steel wire. Azimuthal positions of this target were accurately estimated from the passive images. Next, stable and inertial cavitation was passively imaged in saline solution sonicated at 520 kHz. Bubble clusters formed in the saline samples were consistently located on both passive images and B-scans. Passive images were also created using broadband emissions from bovine liver sonicated at 2.2 MHz. Agreement was found between the images and source beam shape, indicating an ability to map therapeutic ultrasound beams in situ. The relation between these broadband emissions, sonication amplitude, and exposure conditions are discussed. PMID- 20000922 TI - Automatic estimation of position and orientation of an acoustic source by a microphone array network. AB - A method which automatically provides the position and orientation of a directional acoustic source in an enclosed environment is proposed. In this method, different combinations of the estimated parameters from the received signals and the microphone positions of each array are used as inputs to the artificial neural network (ANN). The estimated parameters are composed of time delay estimates (TDEs), source position estimates, distance estimates, and energy features. The outputs of the ANN are the source orientation (one out of four possible orientations shifted by 90 degrees and either the best array which is defined as the nearest to the source) or the source position in two dimensional/three dimensional (2D/3D) space. This paper studies the position and orientation estimation performances of the ANN for different input/output combinations (and different numbers of hidden units). The best combination of parameters (TDEs and microphone positions) yields 21.8% reduction in the average position error compared to the following baselines and a correct orientation ratio greater than 99%. Position localization baselines consist of a time delay of arrival based method with an average position error of 34.1 cm and the steered response power with phase transform method with an average position error of 29.8 cm in 3D space. PMID- 20000923 TI - Exploitation of symmetries for image reconstruction in linearized variable density diffraction tomography. AB - The problem of reconstructing an object's weakly varying compressibility and density distributions in three-dimensional (3D) acoustic diffraction tomography is studied. Based on the Fourier diffraction projection theorem for acoustic media, it is demonstrated that the 3D Fourier components of an object's compressibility and density distributions can be decoupled algebraically, thereby providing a method for separately reconstructing the distributions. This is facilitated by the identification and exploitation of tomographic symmetries and the rotational invariance of the imaging model. The developed reconstruction methods are investigated by use of computer- simulation studies. The application of the proposed image reconstruction strategy to other tomography problems is discussed. PMID- 20000924 TI - Adaptive extraction of modulation for cavitation noise. AB - Modulation analysis is an important issue in target classification and identification for ship-radiated noise. However, the modulated cavitation noise sought for analyzing is always submerged under strong ambient noise and difficult to be separated out. In this paper, an approach is proposed to extract the modulated cavitation noise adaptively by combining empirical mode decomposition and singular value decomposition. The results for both synthetical and practical signals demonstrate the practicability and effectivity of the approach. PMID- 20000925 TI - Comparison of in-situ calibration methods for quantifying input to the middle ear. AB - Sound pressure level in-situ measurements are sensitive to standing-wave pressure minima and have the potential to result in over-amplification with risk to residual hearing in hearing-aid fittings. Forward pressure level (FPL) quantifies the pressure traveling toward the tympanic membrane and may be a potential solution as it is insensitive to ear-canal pressure minima. Derivation of FPL is dependent on a Thevenin-equivalent source calibration technique yielding source pressure and impedance. This technique is found to accurately decompose cavity pressure into incident and reflected components in both a hard-walled test cavity and in the human ear canal through the derivation of a second sound-level measure termed integrated pressure level (IPL). IPL is quantified by the sum of incident and reflected pressure amplitudes. FPL and IPL were both investigated as measures of sound-level entering the middle ear. FPL may be a better measure of middle-ear input because IPL is more dependent on middle-ear reflectance and ear-canal conductance. The use of FPL in hearing-aid applications is expected to provide an accurate means of quantifying high-frequency amplification. PMID- 20000926 TI - Postnatal development of sound pressure transformations by the head and pinnae of the cat: Binaural characteristics. AB - There are three acoustical cues to sound location: Interaural time differences (ITDs), interaural level differences (ILDs), and monaural spectral shape cues. During development, the increasing interaural distance and pinnae size associated with a growing head and pinnae result in localization cues that change continuously until maturation is complete. Here the authors report measurements of both the physical dimensions of the head and pinnae, as well as acoustical measurements of the binaural localization cues of cats aged 1.3 weeks to adulthood. For a given source location, ILD magnitude tended to increase with both frequency and age. Moreover, the range of significant ILD production (approximately 10 dB) shifted with age from higher to lower frequencies. ITD magnitude increased with age. Partial correlation analyses revealed that increasing pinnae size accounted for approximately 31% of the variance in the development of ILDs while increasing head size accounted for virtually none. On the other hand, increases in both the head and pinnae sizes contributed to the development of the ITD cues accounting for approximately 71% and approximately 25% of the variance, respectively. ILD and ITD cues in cats reach maturity by approximately 16 and approximately 22 weeks, respectively, which match the time period over which the pinnae and head dimensions reach maturity. PMID- 20000927 TI - Use of the matching pursuit algorithm with a dictionary of asymmetric waveforms in the analysis of transient evoked otoacoustic emissions. AB - Transiently evoked otoacoustic emissions (TEOAEs) are normally modeled as the sum of asymmetric waveforms. However, some previous studies of TEOAEs used time frequency (TF) methods to decompose the signals into symmetric waveforms. This approach was justified mainly as a means to reduce the complexity of the calculations. The present study extended the dictionary of numeric functions to incorporate asymmetric waveforms into the analysis. The necessary calculations were carried out using an adaptive approximation algorithm based on the matching pursuit (MP) numerical technique. The classic MP dictionary uses Gabor functions and consists of waveforms described by five parameters, namely, frequency, latency, time span, amplitude, and phase. In the present investigation, a sixth parameter, the degree of asymmetry, was added in order to enhance the flexibility of this approach. The effects of expanding the available functions were evaluated by means of both simulations using synthetic signals and authentic TEOAEs. The resulting analyses showed that the contributions of asymmetric components in the OAE signal are appreciable. In short, the expanded analysis method brought about important improvements in identifying TEOAE components including the correct detection of components with long decays, which are often related to spontaneous OAE activity, the elimination of a "dark energy" effect in TF distributions, and more reliable estimates of latency-frequency relationships. The latter feature is especially important for correct estimation of latency-frequency data, which is a crucial factor in investigations of OAE-generation mechanisms. PMID- 20000928 TI - Informational factors in identifying environmental sounds in natural auditory scenes. AB - In a non-linguistic analog of the "cocktail-party" scenario, informational and contextual factors were found to affect the recognition of everyday environmental sounds embedded in naturalistic auditory scenes. Short environmental sound targets were presented in a dichotic background scene composed of either a single stereo background scene or a composite background scene created by playing different background scenes to the different ears. The side of presentation, time of onset, and number of target sounds were varied across trials to increase the uncertainty for the participant. Half the sounds were contextually congruent with the background sound (i.e., consistent with the meaningful real-world sound environment represented in the auditory scene) and half were incongruent. The presence of a single competing background scene decreased identification accuracy, suggesting an informational masking effect. In tandem, there was a contextual pop-out effect, with contextually incongruent sounds identified more accurately. However, when targets were incongruent with the real-world context of the background scene, informational masking was reduced. Acoustic analyses suggested that this contextual pop-out effect was driven by a mixture of perceptual differences between the target and background, as well as by higher level cognitive factors. These findings indicate that identification of environmental sounds in naturalistic backgrounds is an active process that requires integrating perceptual, attentional, and cognitive resources. PMID- 20000929 TI - Perceptual and emotional categorization of sound. AB - This paper investigates how different types of data from psychoacoustical experiments may be combined to render further knowledge about the mechanisms underlying sound perception. Two studies were conducted with auditory alerts of short duration. First, an experiment where participants rated the dissimilarity among the auditory alerts was performed. This resulted in a two-dimensional multi dimensional scaling solution. Second, an experiment where participants evaluated the stimuli with semantic descriptors and rated their emotional reactions to the sounds was performed. The output of this experiment was a reduced set of underlying perceptual and emotional dimensions. The results of the two experiments were then integrated by the use of multi-dimensional perceptual unfolding and a set mediation analyses. The integrative analyses showed that part of the cognitive categorization of the semantic descriptors was mediated by the emotional reactions to the sounds. The results are discussed in relation to theories of auditory perception and emotional response categorization. PMID- 20000930 TI - A comparison of the temporal weighting of annoyance and loudness. AB - The influence of single temporal portions of a sound on global annoyance and loudness judgments was measured using perceptual weight analysis. The stimuli were 900-ms noise samples randomly changing in level every 100 ms. For loudness judgments, Pedersen and Ellermeier [J. Acoust. Soc. Am. 123, 963-972 (2008)] found that listeners attach greater weight to the beginning and ending than to the middle of a stimulus. Qualitatively similar weights were expected for annoyance. Annoyance and loudness judgments were obtained from 12 listeners in a two-interval forced-choice task. The results demonstrated a primacy effect for the temporal weighting of both annoyance and loudness. However, a significant recency effect was observed only for annoyance. Potential explanations of these weighting patterns are discussed. Goodness-of-fit analysis showed that the prediction of annoyance and loudness can be improved by allowing a non-uniform weighting of single temporal portions of the signal, rather than assuming a uniform weighting as in measures like the energy-equivalent level (L(eq)). A second experiment confirmed that the listeners were capable of separating annoyance and loudness of the stimuli. Noises with the same L(eq) but different amplitude modulation depths were judged to differ in annoyance but not in loudness. PMID- 20000931 TI - What makes a melody: The perceptual singularity of pitch sequences. AB - This study investigated the ability of normal-hearing listeners to process random sequences of tones varying in either pitch or loudness. Same/different judgments were collected for pairs of sequences with a variable length (up to eight elements) and built from only two different elements, which were 200-ms harmonic complex tones. The two possible elements of all sequences had a fixed level of discriminability, corresponding to a d(') value of about 2, irrespective of the auditory dimension (pitch or loudness) along which they differed. This made it possible to assess sequence processing per se, independent of the accuracy of sound encoding. Pitch sequences were found to be processed more effectively than loudness sequences. However, that was the case only when the sequence elements included low-rank harmonics, which could be at least partially resolved in the auditory periphery. The effect of roving and transposition was also investigated. These manipulations reduced overall performance, especially transposition, but an advantage for pitch sequences was still observed. These results suggest that automatic frequency-shift detectors, available for pitch sequences but not loudness sequences, participate in the effective encoding of melodies. PMID- 20000932 TI - The role of across-frequency processes in dichotic listening conditions. AB - In the bandwidening experiment with a diotic noise masker, an apparently wider critical bandwidth has often been reported when a dichotic signal (Spi) is used instead of a diotic signal (So). Two competing across-channel processes were proposed to account for this apparently wider critical bandwidth: (i) A detrimental across-channel effect reducing the binaural masking-level difference (BMLD) for broadband maskers and (ii) a beneficial integration of information across channels for narrowband maskers. The two hypotheses result in different predictions of the BMLD in the notched-noise experiment: According to the first hypothesis, the change in BMLD with notch width is determined by the level dependence of the BMLD for a narrowband masker centered at the signal frequency, whereas the second hypothesis predicts that it is determined by the level dependence of the BMLD for a broadband masker. To test the hypotheses, masked thresholds of a diotic or dichotic 500-Hz signal were measured for a diotic notched-noise masker as a function of notch width. In addition, thresholds were measured for a diotic broadband and narrowband masker as a function of masker level. The data indicate that neither of the two hypotheses is able to predict the continuous decrease in the BMLD as the notch width increases. PMID- 20000933 TI - Effects of bandwidth on auditory localization with a noise masker. AB - Although high-frequency content is known to be critically important for the accurate location of isolated sounds, relatively little is known about the importance of high-frequency spectral content for the localization of sounds in the presence of a masker. In this experiment, listeners were asked to identify the location of a pulsed-noise target in the presence of a randomly located continuous noise masker. Both the target and masker were low-pass filtered at one of eight cutoff frequencies ranging from 1 to 16 kHz, and the signal-to-noise ratio was varied from -12 to +12 dB. The results confirm the importance of high frequencies for the localization of isolated sounds, and show that high-frequency content remains critical in cases where the target sound is masked by a spatially separated masker. In fact, when two sources of the same level are randomly located in space, these results show that a decrease in stimulus bandwidth from 16 to 12 kHz might result in a 30% increase in overall localization error. PMID- 20000934 TI - Amplification of interaural level differences improves sound localization in acoustic simulations of bimodal hearing. AB - Users of a cochlear implant and contralateral hearing aid are sensitive to interaural level differences (ILDs). However, when using their clinical devices, most of these subjects cannot use ILD cues for localization in the horizontal plane. This is partly due to a lack of high-frequency residual hearing in the acoustically stimulated ear. Using acoustic simulations of a cochlear implant and hearing loss, it is shown that localization performance can be improved by up to 14 degrees rms error relative to 48 degrees rms error for broadband noise by artificially introducing ILD cues in the low frequencies. The algorithm that was used for ILD introduction is described. PMID- 20000935 TI - Using a signal cancellation technique involving impulse response to assess directivity of hearing aids. AB - The directional microphone systems of modern digital hearing aids are capable of changing their spatial directivity pattern and/or the microphone mode in response to changes in the properties of environmental sounds. These adaptive/automatic features make measurement of a hearing aid's directivity in a given test environment very difficult. Assessing the directivity of such systems requires a signal that can record the system's response while not changing the system's directivity. This paper proposes a method using a signal cancellation technique involving impulse responses to acoustically assess a hearing aid's directivity (referred to as the IR method). The impulse is presumed to be undetectable to the adaptive/automatic system because it contains little energy and a short response could be recorded before the system actually reacts. In the current study, the IR method was evaluated by testing five adaptive/automatic directional hearing aids in noise of various intensities. The results revealed that the IR method was an accurate and repeatable way to assess slow-acting directional systems in noise of varying intensities and fast-acting systems in noise of high intensities. PMID- 20000936 TI - Analysis of acoustic reduction using spectral similarity measures. AB - Articulatory and acoustic reduction can manifest itself in the temporal and spectral domains. This study introduces a measure of spectral reduction, which is based on the speech decoding techniques commonly used in automatic speech recognizers. Using data for four frequent Dutch affixes from a large corpus of spontaneous face-to-face conversations, it builds on an earlier study examining the effects of lexical frequency on durational reduction in spoken Dutch [Pluymaekers, M. et al. (2005). J. Acoust. Soc. Am. 118, 2561-2569], and compares the proposed measure of spectral reduction with duration as a measure of reduction. The results suggest that the spectral reduction scores capture other aspects of reduction than duration. While duration can--albeit to a moderate degree--be predicted by a number of linguistically motivated variables (such as word frequency, segmental context, and speech rate), the spectral reduction scores cannot. This suggests that the spectral reduction scores capture information that is not directly accounted for by the linguistically motivated variables. The results also show that the spectral reduction scores are able to predict a substantial amount of the variation in duration that the linguistically motivated variables do not account for. PMID- 20000937 TI - The dynamic range of speech, compression, and its effect on the speech reception threshold in stationary and interrupted noise. AB - Changes in the speech reception threshold (SRT) after amplitude compression of speech or speech in noise may be due to changes in the dynamic range of the speech signal. However, current models set up to predict the speech intelligibility consider the dynamic range of speech to be fixed regardless of the type of compression. The present paper describes two experiments with normal hearing subjects to examine the effect of the dynamic range on the SRT in stationary and interrupted noise after wide dynamic range compression. The dynamic range has been varied by compression or expansion of only the speech signal, leaving the masking noise unaltered, or by compression or expansion of the mixed speech-in-noise signal. The results show that compression affects the SRT, both in a positive or a negative direction, not only due to dynamic range but also due to distortion of the speech signal. PMID- 20000938 TI - Speech identification in noise: Contribution of temporal, spectral, and visual speech cues. AB - This study investigated the degree to which two types of reduced auditory signals (cochlear implant simulations) and visual speech cues combined for speech identification. The auditory speech stimuli were filtered to have only amplitude envelope cues or both amplitude envelope and spectral cues and were presented with/without visual speech. In Experiment 1, IEEE sentences were presented in quiet and noise. For in-quiet presentation, speech identification was enhanced by the addition of both spectral and visual speech cues. Due to a ceiling effect, the degree to which these effects combined could not be determined. In noise, these facilitation effects were more marked and were additive. Experiment 2 examined consonant and vowel identification in the context of CVC or VCV syllables presented in noise. For consonants, both spectral and visual speech cues facilitated identification and these effects were additive. For vowels, the effect of combined cues was underadditive, with the effect of spectral cues reduced when presented with visual speech cues. Analysis indicated that without visual speech, spectral cues facilitated the transmission of place information and vowel height, whereas with visual speech, they facilitated lip rounding, with little impact on the transmission of place information. PMID- 20000939 TI - Effects of electrode separation between speech and noise signals on consonant identification in cochlear implants. AB - The aim of the present study was to examine cochlear implant (CI) users' perceptual segregation of speech from background noise with differing degrees of electrode separation between speech and noise. Eleven users of the nucleus CI system were tested on consonant identification using an experimental processing scheme called "multi-stream processing" in which speech and noise stimuli were processed separately and interleaved. Speech was presented to either ten (every other electrode) or six electrodes (every fourth electrode). Noise was routed to either the same (the "overlapped" condition) or a different set of electrodes (the "interlaced" condition), where speech and noise electrodes were separated by one- and two-electrode spacings for ten- and six-electrode presentations, respectively. Results indicated a small but significant improvement in consonant recognition (5%-10%) in the interlaced condition with a two-electrode spacing (approximately 1.1 mm) in two subjects. It appears that the results were influenced by peripheral channel interactions, partially accounting for individual variability. Although the overall effect was small and observed from a small number of subjects, the present study demonstrated that CI users' performance on segregating the target from the background might be improved if these sounds were presented with sufficient peripheral separation. PMID- 20000940 TI - Automatic detection of the second subglottal resonance and its application to speaker normalization. AB - Speaker normalization typically focuses on inter-speaker variabilities of the supraglottal (vocal tract) resonances, which constitute a major cause of spectral mismatch. Recent studies have shown that the subglottal airways also affect spectral properties of speech sounds, and promising results were reported using the subglottal resonances for speaker normalization. This paper proposes a reliable algorithm to automatically estimate the second subglottal resonance (Sg2) from speech signals. The algorithm is calibrated on children's speech data with simultaneous accelerometer recordings from which Sg2 frequencies can be directly measured. A cross-language study with bilingual Spanish-English children is performed to investigate whether Sg2 frequencies are independent of speech content and language. The study verifies that Sg2 is approximately constant for a given speaker and thus can be a good candidate for limited data speaker normalization and cross-language adaptation. A speaker normalization method using Sg2 is then presented. This method is computationally more efficient than maximum likelihood based vocal tract length normalization (VTLN), with performance better than VTLN for limited adaptation data and cross-language adaptation. Experimental results confirm that this method performs well in a variety of testing conditions and tasks. PMID- 20000941 TI - Surface response of a fractional order viscoelastic halfspace to surface and subsurface sources. AB - Previous studies by the second author published in this journal focused on low audible frequency (40-400 Hz) shear and surface wave motion in and on a viscoelastic material representative of biological tissue. Specific cases considered were that of surface wave motion on a halfspace caused by a finite rigid circular disk located on the surface and oscillating normal to it [Royston et al., J. Acoust. Soc. Am. 106, 3678-3686 (1999)] and compression, shear, and surface wave motion in a halfspace generated by a subsurface finite dipole [Royston et al., J. Acoust. Soc. Am. 113, 1109-1121 (2003)]. In both studies, a Voigt model of viscoelasticity was assumed in the theoretical treatment, which resulted in agreement between theoretical predictions and experimental measurements over a limited frequency range. In the present article, the linear viscoelastic assumption in these two prior works is revisited to consider a (still linear) fractional order Voigt model, where the rate-dependent damping component that is dependent on the first derivative of time is replaced with a component that is dependent on a fractional derivative of time. It is shown that in both excitation source configurations, the fractional order Voigt model assumption improves the match of theory to experiment over a wider frequency range (in some cases up to the measured range of 700 Hz). PMID- 20000942 TI - Predictions of angle dependent tortuosity and elasticity effects on sound propagation in cancellous bone. AB - The anisotropic pore structure and elasticity of cancellous bone cause wave speeds and attenuation in cancellous bone to vary with angle. Previously published predictions of the variation in wave speed with angle are reviewed. Predictions that allow tortuosity to be angle dependent but assume isotropic elasticity compare well with available data on wave speeds at large angles but less well for small angles near the normal to the trabeculae. Claims for predictions that only include angle-dependence in elasticity are found to be misleading. Audio-frequency data obtained at audio-frequencies in air-filled bone replicas are used to derive an empirical expression for the angle-and porosity dependence of tortuosity. Predictions that allow for either angle dependent tortuosity or angle dependent elasticity or both are compared with existing data for all angles and porosities. PMID- 20000943 TI - Frequency dependence of average phase shift from human calcaneus in vitro. AB - If dispersion in a medium is weak and approximately linear with frequency (over the experimental band of frequencies), then it can be shown that the constant term in a polynomial representation of phase shift as a function of frequency can produce errors in measurements of phase-velocity differences in through transmission, substitution experiments. A method for suppressing the effects of the constant phase shift in the context of the single-wave-model was tested on measurements from 30 cancellous human calcaneus samples in vitro. Without adjustment for constant phase shifts, the estimated phase velocity at 500 kHz was 1516+/-6 m/s (mean+/-standard error), and the estimated dispersion was -24+/-4 m/s MHz (mean+/-standard error). With adjustment for constant phase shifts, the estimated mean velocity decreased by 4-9 m/s, and the estimated magnitude of mean dispersion decreased by 50%-100%. The average correlation coefficient between the measured attenuation coefficient and frequency was 0.997+/-0.0026 (mean+/ standard deviation), suggesting that the signal for each sample was dominated by one wave. A single-wave, linearly dispersive model conformed to measured complex transfer functions from the 30 cancellous-bone samples with an average root-mean square error of 1.9%+/-1.0%. PMID- 20000944 TI - Influence of the filling fluid on frequency-dependent velocity and attenuation in cancellous bones between 0.35 and 2.5 MHz. AB - The paper is focused on experiments on human cancellous bones filled with different fluids with the goal of evaluating their contribution to velocity dispersion, absorption, and scattering mechanisms. The specimens were measured first filled with marrow and subsequently, after marrow removal, with water and alcohol. No significant influence of the fluids was evidenced on the attenuation coefficient. Given the absence of impact of viscosity of the saturating fluid, the authors hypothesized that the source of attenuation is associated with viscoelastic absorption in the solid trabeculae and with scattering. Alteration of scattering obtained by changing the acoustic impedance mismatch between the fluid (alcohol vs water) and the trabeculae was reflected neither in the attenuation nor in its slope. This led the authors to suggest that longitudinal to-shear scattering together with absorption in the solid phase are candidates as main sources for the attenuation. The differences in velocity values indicate that the elastic properties of the fluid are main determinants of the phase velocity. This finding is particularly significant in the context of /in vivo/ measurements, because it demonstrates that the subject-dependent properties of marrow may partly explain the inter-subject variability of speed of sound values. PMID- 20000945 TI - Pinna-rim skin folds narrow the sonar beam in the lesser false vampire bat (Megaderma spasma). AB - False vampire bats (genus Megaderma) employ active as well as passive biosonars. In the present work, the acoustic impact of a conspicuous feature of the animals' ear morphology, skin folds of the pinna rim linking the two pinnae at the midline, has been studied using a numerical approach. Automated methods have been devised to measure the largest width of the beam patterns irrespective of beam orientation. A total of six pinna shapes from three individuals have been studied. For all these shapes, it was found that the reception biosonar beams had approximately elliptic cross-sections with the largest beamwidth being on average almost twice as large as the beamwidth in the orthogonal direction. The directions of the largest beamwidths were scattered around the azimuthal dimension. Removal of the skin folds resulted in significant widening of the beams along their widest dimensions with an increase in beamwidth of 9.2 degrees (a 30% change) on average. The strength and repeatability of this effect across individuals suggest the hypothesis that skin folds are functionally relevant to the animals' biosonar system. It may be a morphological adaptation to biosonar tasks that benefit from a narrow beam such as the detection of faint sounds or precise localization. PMID- 20000946 TI - Bowhead whale (Balaena mysticetus) songs in the Chukchi Sea between October 2007 and May 2008. AB - This paper reports on the acoustic detection of bowhead whale (Balaena mysticetus) songs from the Bering-Chukchi-Beaufort stock, including the first recordings of songs in the fall and early winter. Bowhead whale songs were detected almost continuously in the Chukchi Sea between October 30, 2007 and January 1, 2008 and twice from April 16 to May 5, 2008 during a long-term deployment of five acoustic recorders moored off Point Lay and Wainwright, AK, between October 21, 2007 and August 3, 2008. Two complex and four simple songs were detected. The complex songs consisted of highly stereotyped sequences of four units. The simple songs were primarily made of sequences of two to three moan types whose repetition patterns were constant over short periods but more variable over time. Multiple song types were recorded simultaneously and there is evidence of synchronized song variation over time. The implications of the spatiotemporal distribution of song detection with respect to the migratory and mating behavior of western Arctic bowheads are discussed. PMID- 20000947 TI - Acoustic analysis of primate air sacs and their effect on vocalization. AB - This paper presents an analysis of the acoustic impedance of primate air sacs and their interaction with the vocal tract. A lumped element model is derived and it is found that the inertance of the neck and the volume of the air sac are relevant, as well as the mass and stiffness of the walls (depending on the tissue). It is also shown that at low frequencies, radiation from the air sac can be non-negligible, even if the mouth is open. It is furthermore shown that an air sac can add one or two low resonances to the resonances of the oral tract, and that it shifts up the oral tract's resonances below approximately 2000 Hz, and shifts them closer together. The theory was verified by acoustic measurements and applied to the red howler monkey (Alouatta seniculus) and the siamang (Symphalangus syndactylus). The theory describes the physical models and the siamang calls correctly, but appears incomplete for the howler monkey vocalizations. The relation between air sacs and the evolution of speech is discussed briefly, and it is proposed that an air sac would reduce the ability to produce distinctive speech, but would enhance the impression of size of the vocalizer. PMID- 20000948 TI - Effect of ultrasound contrast agent dose on the duration of focused-ultrasound induced blood-brain barrier disruption. AB - It has been shown that focused ultrasound (FUS) is capable of noninvasive and reversible disruption of the blood-brain barrier (BBB) at target regions when applied in the presence of ultrasound contrast agent (UCA). The purpose of this study was to investigate the effects of UCA dose on the reversibility of BBB disruption induced by the same acoustical power of FUS. Sonications were applied at an ultrasound frequency of 1 MHz with a 5% duty cycle and a repetition frequency of 1 Hz. The brains of 66 male Sprague-Dawley rats were subjected to sonications with three doses of UCA. BBB integrity was evaluated via femoral vein injection of Evans Blue (EB) while the rats were anesthetized. The relationship between UCA dose and the region of EB extravasation was evaluated at an acoustic power of 1.43 W. The BBB disruption, as indexed by the amount of EB extravasation, was the largest immediately after the sonications. The quantity of Evans blue extravasation decreased as a function of time at various UCA doses. This study demonstrates that the appropriate dose of UCA not only enhance the BBB opening but also effectively aid in controlling the duration of BBB disruption. PMID- 20000949 TI - Ambient pressure sensitivity of microbubbles investigated through a parameter study. AB - Measurements on microbubbles clearly indicate a relation between the ambient pressure and the acoustic behavior of the bubble. The purpose of this study was to optimize the sensitivity of ambient pressure measurements, using the subharmonic component, through microbubble response simulations. The behavior of two microbubbles corresponding to two different contrast agents was investigated as a function of driving pulse and ambient overpressure p(ov). Simulations of Levovist using a rectangular driving pulse show an almost linear reduction in the subharmonic component as p(ov) is increased. For a 20 cycle driving pulse, a reduction of 4.6 dB is observed when changing p(ov) from 0 to 25 kPa. Increasing the pulse duration makes the reduction even clearer. For a pulse with 64 cycles, the reduction is 9.9 dB. This simulation is in good correspondence with measurement results presented in the literature. Further simulations of Levovist show that also the shape and the acoustic pressure of the driving pulse are very important factors. The best pressure sensitivity of Levovist was found to be 0.88 dB/kPa. For Sonazoid, a sensitivity of 1.14 dB/kPa has been found, although the reduction is not completely linear as a function of the ambient pressure. PMID- 20001013 TI - A robust approach to calculate entropy change based on density functional theory in the energy representation. AB - We have developed a new approach to accurately calculate entropy change based on density functional theory in the energy representation. The entropy change was evaluated using the derived equation and energy distributions computed using molecular simulation and reweighting techniques. This approach was applied to a harmonic oscillator, an alanine dipeptide, and a small protein. We found that the results were accurate compared to conventional approaches, such as the quasiharmonic approximation. PMID- 20001014 TI - Modeling the anisotropic self-assembly of spherical polymer-grafted nanoparticles. AB - Recent experimental results demonstrated that polymer grafted nanoparticles in solvents display self-assembly behavior similar to the microphase separation of block copolymers and other amphiphiles. We present a mean-field theory and complementary computer simulations to shed light on the parametric underpinnings of the experimental observations. Our theory suggests that such self-assembled structures occur most readily when the nanoparticle size is comparable to the radius of gyration of the polymer brush chains. Much smaller particle sizes are predicted to yield uniform particle dispersions, while larger particles are expected to agglomerate due to phase separation from the solvent. Selected aspects of our theoretical predictions are corroborated by computer simulations. PMID- 20001015 TI - Controlling the composition of a confined fluid by an electric field. AB - Starting from a generic model of a pore/bulk mixture equilibrium, we propose a novel method for modulating the composition of the confined fluid without having to modify the bulk state. To achieve this, two basic mechanisms-sensitivity of the pore filling to the bulk thermodynamic state and electric field effect-are combined. We show by Monte Carlo simulation that the composition can be controlled both in a continuous and in a jumpwise way. Near the bulk demixing instability, we demonstrate a field induced population inversion in the pore. The conditions for the realization of this method should be best met with colloids, but being based on robust and generic mechanisms, it should also be applicable to some molecular fluids. PMID- 20001016 TI - O+OH-->O(2)+H: A key reaction for interstellar chemistry. New theoretical results and comparison with experiment. AB - We report extensive, fully quantum, time-independent (TID) calculations of cross sections at low collision energies and rate constants at low temperatures for the O+OH reaction, of key importance in the production of molecular oxygen in cold, dark, interstellar clouds and in the chemistry of the Earth's atmosphere. Our calculations are compared with TID calculations within the J-shifting approximation, with wave-packet calculations, and with quasiclassical trajectory calculations. The fully quantum TID calculations yield rate constants higher than those from the more approximate methods and are qualitatively consistent with a low-temperature extrapolation of earlier experimental values but not with the most recent experiments at the lowest temperatures. PMID- 20001017 TI - State-selective spectroscopy of water up to its first dissociation limit. AB - A joint experimental and first-principles quantum chemical study of the vibration rotation states of the water molecule up to its first dissociation limit is presented. Triple-resonance, quantum state-selective spectroscopy is used to probe the entire ladder of water's stretching vibrations up to 19 quanta of OH stretch, the last stretching state below dissociation. A new ground state potential energy surface of water is calculated using a large basis set and an all-electron, multireference configuration interaction procedure, which is augmented by relativistic corrections and fitted to a flexible functional form appropriate for a dissociating system. Variational nuclear motion calculations on this surface are used to give vibrational assignments. A total of 44 new vibrational states and 366 rotation-vibration energy levels are characterized; these span the region from 35,508 to 41,126 cm(-1) above the vibrational ground state. PMID- 20001018 TI - Using wave-packet interferometry to monitor the external vibrational control of electronic excitation transfer. AB - We investigate the control of electronic energy transfer in molecular dimers through the preparation of specific vibrational coherences prior to electronic excitation, and its observation by nonlinear wave-packet interferometry (nl-WPI). Laser-driven coherent nuclear motion can affect the instantaneous resonance between site-excited electronic states and thereby influence short-time electronic excitation transfer (EET). We first illustrate this control mechanism with calculations on a dimer whose constituent monomers undergo harmonic vibrations. We then consider the use of nl-WPI experiments to monitor the nuclear dynamics accompanying EET in general dimer complexes following impulsive vibrational excitation by a subresonant control pulse (or control pulse sequence). In measurements of this kind, two pairs of polarized phase-related femtosecond pulses following the control pulse generate superpositions of coherent nuclear wave packets in optically accessible electronic states. Interference contributions to the time- and frequency-integrated fluorescence signals due to overlaps among the superposed wave packets provide amplitude-level information on the nuclear and electronic dynamics. We derive the basic expression for a control-pulse-dependent nl-WPI signal. The electronic transition moments of the constituent monomers are assumed to have a fixed relative orientation, while the overall orientation of the complex is distributed isotropically. We include the limiting case of coincident arrival by pulses within each phase-related pair in which control-influenced nl-WPI reduces to a fluorescence-detected pump-probe difference experiment. Numerical calculations of pump-probe signals based on these theoretical expressions are presented in the following paper [J. D. Biggs and J. A. Cina, J. Chem. Phys. 131, 224302 (2009)]. PMID- 20001019 TI - Quantum algorithm for molecular properties and geometry optimization. AB - Quantum computers, if available, could substantially accelerate quantum simulations. We extend this result to show that the computation of molecular properties (energy derivatives) could also be sped up using quantum computers. We provide a quantum algorithm for the numerical evaluation of molecular properties, whose time cost is a constant multiple of the time needed to compute the molecular energy, regardless of the size of the system. Molecular properties computed with the proposed approach could also be used for the optimization of molecular geometries or other properties. For that purpose, we discuss the benefits of quantum techniques for Newton's method and Householder methods. Finally, global minima for the proposed optimizations can be found using the quantum basin hopper algorithm, which offers an additional quadratic reduction in cost over classical multi-start techniques. PMID- 20001020 TI - Accurate yet feasible post-Hartree-Fock computation of magnetic interactions in large biradicals through a combined variational/perturbative approach: Setup and validation. AB - We present a scheme for the calculation of the spin-spin coupling term J in diradicals which is quantitatively accurate and computationally cheap. The method exploits the use of modified virtual orbitals and perturbation theory, incorporated in a multireference configuration interaction approach. The results obtained for model diradical species which exhibit ferromagnetic and antiferromagnetic coupling are fully satisfactory and very promising for future applications of the method to larger molecular systems of technological interest in magnetic-based devices. PMID- 20001021 TI - Density functional method including weak interactions: Dispersion coefficients based on the local response approximation. AB - A new method to calculate the atom-atom dispersion coefficients in a molecule is proposed for the use in density functional theory with dispersion (DFT-D) correction. The method is based on the local response approximation due to Dobson and Dinte [Phys. Rev. Lett. 76, 1780 (1996)], with modified dielectric model recently proposed by Vydrov and van Voorhis [J. Chem. Phys. 130, 104105 (2009)]. The local response model is used to calculate the distributed multipole polarizabilities of atoms in a molecule, from which the dispersion coefficients are obtained by an explicit frequency integral of the Casimir-Polder type. Thus obtained atomic polarizabilities are also used in the damping function for the short-range singularity. Unlike empirical DFT-D methods, the local response dispersion (LRD) method is able to calculate the dispersion energy from the ground-state electron density only. It is applicable to any geometry, free from physical constants such as van der Waals radii or atomic polarizabilities, and computationally very efficient. The LRD method combined with the long-range corrected DFT functional (LC-BOP) is applied to calculations of S22 weakly bound complex set [Phys. Chem. Chem. Phys. 8, 1985 (2006)]. Binding energies obtained by the LC-BOP+LRD agree remarkably well with ab initio references. PMID- 20001022 TI - Exact Kohn-Sham potential of strongly correlated finite systems. AB - The dissociation of molecules, even the most simple hydrogen molecule, cannot be described accurately within density functional theory because none of the currently available functionals accounts for strong on-site correlation. This problem led to a discussion of properties that the local Kohn-Sham potential has to satisfy in order to correctly describe strongly correlated systems. We derive an analytic expression for the nontrivial form of the Kohn-Sham potential in between the two fragments for the dissociation of a single bond. We show that the numerical calculations for a one-dimensional two-electron model system indeed approach and reach this limit. It is shown that the functional form of the potential is universal, i.e., independent of the details of the two fragments. PMID- 20001023 TI - Calculations of rovibrational energies and dipole transition intensities for polyatomic molecules using MULTIMODE. AB - We report rigorous calculations of rovibrational energies and dipole transition intensities for three molecules using a new version of the code MULTIMODE. The key features of this code which permit, for the first time, such calculations for moderately sized but otherwise general polyatomic molecules are briefly described. Calculations for the triatomic molecule BF(2) are done to validate the code. New calculations for H(2)CO and H(2)CS are reported; these make use of semiempirical potentials but ab initio dipole moment surfaces. The new dipole surface for H(2)CO is a full-dimensional fit to the dipole moment obtained with the coupled-cluster with single and double excitations and a perturbative treatment of triple excitations method with the augmented correlation consistent triple zeta basis set. Detailed comparisons are made with experimental results from a fit to relative data for H(2)CS and absolute intensities from the HITRAN database for H(2)CO. PMID- 20001024 TI - Gaussian approximation for the structure function in semiclassical forward backward initial value representations of time correlation functions. AB - Initial value representations (IVRs) of semiclassical (SC) theory provide a general approach for adding quantum mechanical effects to classical molecular dynamics simulations of large molecular systems. Of the various versions of SC IVR methodology for evaluating time correlation functions, the Fourier transform forward-backward (FB) approach is the simplest one that is able to describe true quantum coherence effects, so it is of considerable importance to find efficient and systematic ways for implementing it. It is shown in this paper that a Gaussian approximation for the "structure function"-the dependence of the correlation function on the (typically) momentum jump parameter-provides an efficient and accurate way for doing so. The approach is illustrated by an application to the time-dependent radial distribution function of I(2) (after photoexcitation) in a cluster of (up to 16) argon atoms. PMID- 20001025 TI - Dispersion phenomena in helical flow in a concentric annulus. AB - We examined dispersion phenomena of solutes in helical flow in a concentric annulus through a multiscale approach. The helical flow was developed by the combination of the Poiseuille flow and Couette flow. Here, we present an analytic model that can address the multidimensional Taylor dispersion in the helical flow under a lateral field of thermophoresis (or thermal diffusion) in the gapwise direction. Macroscopic parameters including the average solute velocity and dispersivity were analyzed using relevant microscopic physicochemical properties. The mathematically obtained results were validated by the numerical simulation carried out in this study. The findings show that macrotransport processes are robust and straightforward to handle multidimensional dispersion phenomena of solutes in helical flow. This study is expected to provide a theoretical platform for applications of helical flow such as tube exchangers, oil drilling, and multidimensional field flow fractionations (e.g., helical flow field flow fractionation). PMID- 20001026 TI - Multiconfigurational time-dependent Hartree calculations for tunneling splittings of vibrational states: Theoretical considerations and application to malonaldehyde. AB - Full-dimensional multiconfigurational time-dependent Hartree calculations on the tunneling splitting of the vibrational ground state and the low lying excited states of malonaldehyde are presented. Methodological developments utilizing the symmetry of double well systems for the efficient calculation of tunneling splittings are described and discussed. Important aspects of the theory underlying the previously communicated results for the ground state tunneling splitting [M. D. Coutinho-Neto et al., J. Chem. Phys. 121, 9207 (2004)] are detailed and further developments facilitating the calculation of tunneling splittings for vibrationally excited states are introduced. Utilizing these developments, the 14 lowest vibrational states of malonaldehyde, i.e., seven tunneling splittings, have been computed. The tunneling splittings are found to vary significantly depending on the particular vibrational excitation. This results in a complex pattern of vibrational levels. Studying the dependence of the tunneling splittings on the vibrational excitation, good agreement with available experimental results is found and intuitive interpretations of the results can be given. PMID- 20001027 TI - One-dimensional description of diffusion in a tube of abruptly changing diameter: Boundary homogenization based approach. AB - Reduction of three-dimensional (3D) description of diffusion in a tube of variable cross section to an approximate one-dimensional (1D) description has been studied in detail previously only in tubes of slowly varying diameter. Here we discuss an effective 1D description in the opposite limiting case when the tube diameter changes abruptly, i.e., in a tube composed of any number of cylindrical sections of different diameters. The key step of our approach is an approximate description of the particle transitions between the wide and narrow parts of the tube as trapping by partially absorbing boundaries with appropriately chosen trapping rates. Boundary homogenization is used to determine the trapping rate for transitions from the wide part of the tube to the narrow one. This trapping rate is then used in combination with the condition of detailed balance to find the trapping rate for transitions in the opposite direction, from the narrow part of the tube to the wide one. Comparison with numerical solution of the 3D diffusion equation allows us to test the approximate 1D description and to establish the conditions of its applicability. We find that suggested 1D description works quite well when the wide part of the tube is not too short, whereas the length of the narrow part can be arbitrary. Taking advantage of this description in the problem of escape of diffusing particle from a cylindrical cavity through a cylindrical tunnel we can lift restricting assumptions accepted in earlier theories: We can consider the particle motion in the tunnel and in the cavity on an equal footing, i.e., we can relax the assumption of fast intracavity relaxation used in all earlier theories. As a consequence, the dependence of the escape kinetics on the particle initial position in the system can be analyzed. Moreover, using the 1D description we can analyze the escape kinetics at an arbitrary tunnel radius, whereas all earlier theories are based on the assumption that the tunnel is narrow. PMID- 20001028 TI - Approximate inclusion of quantum effects in transition path sampling. AB - We propose a method for incorporating nuclear quantum effects in transition path sampling studies of systems that consist of a few degrees of freedom that must be treated quantum mechanically, while the rest are classical-like. We used the normal mode centroid method to describe the quantum subsystem, which is a method that is not CPU intensive but still reasonably accurate. We applied this mixed centroid/classical transition path sampling method to a model system that has nontrivial quantum behavior, and showed that it can capture the correct quantum dynamical features. PMID- 20001029 TI - Estimating diffusivity along a reaction coordinate in the high friction limit: Insights on pulse times in laser-induced nucleation. AB - In the high friction limit of Kramers' theory, the diffusion coefficient for motion along the reaction coordinate is a crucial parameter in determining reaction rates from mean first passage times. The Einstein relation between mean squared displacement, time, and diffusivity is inaccurate at short times because of ballistic motion and inaccurate at long times because trajectories drift away from maxima in the potential of mean force. Starting from the Smoluchowski equation for a downward parabolic barrier, we show how drift induced by the potential of mean force can be included in estimating the diffusivity. A modified relation between mean squared displacement, time, and diffusivity now also includes a dependence on the barrier curvature. The new relation provides the diffusivity at the top of the barrier from a linear regression that is analogous to the procedure commonly used with Einstein's relation. The new approach has particular advantages over previous approaches when evaluations of the reaction coordinate are costly or when the reaction coordinate cannot be differentiated to compute restraining forces or velocities. We use the new method to study the dynamics of barrier crossing in a Potts lattice gas model of nucleation from solution. Our analysis shows that some current hypotheses about laser-induced nucleation mechanisms lead to a nonzero threshold laser pulse duration below which a laser pulse will not affect nucleation. We therefore propose experiments that might be used to test these hypotheses. PMID- 20001030 TI - How long are the ends of polyene chains? AB - In this work we study conjugation in all-trans polyene chains H[Single Bond](HC[Double Bond]CH)(n)[Single Bond]H with a view to establishing the length scale for the interaction between conjugated double bonds. As a polyene oligomer is made longer, bond length alternation between formal carbon-carbon single and double bonds diminishes toward the middle of the chain, eventually reaching a constant value characteristic of an "infinite" chain. However those bonds near the end of the chain continue to be influenced by the end, even in the long-chain limit. We have determined optimized geometries for polyene oligomers with up to n=11 repeat units at the MP2/cc-pVTZ level. At this length the central-most bonds are almost converged to the long chain limit, for which we estimate R(C[Double Bond]C)=1.3652 A and R(C[Single Bond]C)=1.4238 A. In contrast, the endmost double bond has a length of 1.3442 A and the endmost single bond has a length of 1.4425 A. We find that a given bond is significantly influenced by conjugation paths through up to six neighboring conjugated double bonds. End effects can also be monitored by examining the energy increment per added monomer as the oligomer length is increased. This analysis also indicates that significant conjugation effects extend out through approximately six neighboring double bonds. From the energy per monomer of the longest chains we extract a value of about 8 kcal/mol for the extra stabilization energy per monomer due to conjugation in long chains. PMID- 20001031 TI - Calculations of nonlinear wave-packet interferometry signals in the pump-probe limit as tests for vibrational control over electronic excitation transfer. AB - The preceding paper [J. D. Biggs and J. A. Cina, J. Chem. Phys. 131, 224101 (2009)] (referred to here as Paper 1), describes a strategy for externally influencing the course of short-time electronic excitation transfer (EET) in molecular dimers and observing the process by nonlinear wave-packet interferometry (nl-WPI). External influence can, for example, be exerted by inducing coherent intramolecular vibration in one of the chromophores prior to short-pulse electronic excitation of the other. Within a sample of isotropically oriented dimers having a specified internal geometry, a vibrational mode internal to the acceptor chromophore can be preferentially driven by electronically nonresonant impulsive stimulated Raman (or resonant infrared) excitation with a short polarized "control" pulse. A subsequent electronically resonant polarized pump then preferentially excites the donor, and EET ensues. Paper 1 investigates control-pulse-influenced nl-WPI as a tool for the spectroscopic evaluation of the effect of coherent molecular vibration on excitation transfer, presenting general expressions for the nl-WPI difference signal from a dimer following the action of a control pulse of arbitrary polarization and shape. Electronic excitation is to be effected and its interchromophore transfer monitored by resonant pump and probe "pulses," respectively, each consisting of an optical-phase-controlled ultrashort pulse-pair having arbitrary polarization, duration, center frequency, and other characteristics. Here we test both the control strategy and its spectroscopic investigation-with some sacrifice of amplitude-level detail-by calculating the pump-probe difference signal. That signal is the limiting case of the control-influenced nl-WPI signal in which the two pulses in the pump pulse pair coincide, as do the two pulses in the probe pulse-pair. We present calculated pump-probe difference signals for (1) a model excitation-transfer complex in which two equal-energy monomers each support one moderately Franck Condon active intramolecular vibration; (2) a simplified model of the covalent dimer dithia-anthracenophane, representing its EET dynamics following selective impulsive excitation of the weakly Franck-Condon active nu(12) anthracene vibration at 385 cm(-1); and (3) a model complex featuring moderate electronic vibrational coupling in which the site energy of the acceptor chromophore is lower than that of the donor. PMID- 20001032 TI - Rotational effects in complex-forming bimolecular substitution reactions: A quantum-mechanical approach. AB - The quantum dynamics of the complex-forming S(N)2 reaction Cl(-)+CH(3)Br- >ClCH(3)+Br(-) is studied with emphasis on rotational effects. The pseudotriatomic system Cl-Me-Br is treated with a corresponding three-dimensional (3D) potential energy surface as a function of the two scattering coordinates and the enclosed angle where the geometry of the methyl group Me is optimized at each point. The 3D space is divided into three different parts, the interaction region, an intermediate region, and the asymptotic region. In line with simple classical-mechanical arguments and previous classical trajectory calculations, initial rotational motion of CH(3)Br seemingly decreases the reaction probability. However, the dynamical inclusion of the rotational degree of freedom and the presence of the many rovibrational product states overall lead to a large increase in reactivity compared to our previous collinear study on this reaction. If the reactant is rotationally excited, the higher vibrational product states are depleted in favor of lower-lying levels. Starting the reaction with rotationless reactants may end up in significant rotational excitation in the product molecules (translation-to-rotation energy transfer). On the other hand, initial rotational energy in rotationally highly excited reactants is to a large amount converted into translational and vibrational energy. The average amount of rotational energy in the products shows a twofold vibrational excitation independent saturation (i.e., memorylessness), with respect to both initial rotational excitation and translational energy. Since only about one-half of all reactant states end in rotationless products, the reaction probability should be increased by a factor of 2; the actually larger reactivity points to other dynamical effects that play an important role in the reaction. PMID- 20001033 TI - Structure and properties of the Zn(+)-D(2) complex. AB - The infrared spectrum of the (66)Zn(+)-D(2) complex is measured in the D-D stretch region (2815-2866 cm(-1)) by detecting Zn(+) photofragments. The spectrum is consistent with the Zn(+)-D(2) complex consisting of a slightly distorted D(2) molecule attached to a ground state Zn(+) ion in a T-shaped equilibrium configuration. From the rotational constants, the vibrationally averaged intermolecular bond length is deduced to be 2.32 A, contracting by 0.02 A upon excitation of the D-D stretch vibrational mode. The band center of the D-D stretch transition is shifted by -154.8 cm(-1) from the Q(0) (1) transition of the free D(2) molecule. Density functional theory calculations are performed to elucidate the molecular bonding in the complex. The current spectroscopic and calculated data for Zn(+)-D(2), together with the previously determined binding energy for Zn(+)-H(2) [1310 cm(-1); P. Weis, et al., J. Phys. Chem. A 101, 2809 (1997)], result in a comprehensive characterization of the Zn(+)-D(2) and Zn(+) H(2) complexes. PMID- 20001034 TI - Studies of the fragmentation of the monocation and dication of methanol. AB - Relative partial ionization cross sections and precursor-specific relative partial ionization cross sections for fragment ions formed by electron ionization of methanol have been measured using time-of-flight mass spectrometry coupled with a two-dimensional ion coincidence technique. Relative cross sections are reported for ionizing energies from 30 to 200 eV. Good agreement is found between our data and one set of recently published absolute partial ionization cross sections. Conversely, discrepancies are observed with another set of recently published data; we attribute these discrepancies to the loss of translationally energetic fragment ions. Our precursor-specific cross sections allow the contribution from single and double ionization to the individual fragment ion yields, following ionization of methanol, to be quantified for the first time. Our analysis shows that the contribution of double ionization to the total ion yield reaches a maximum of 20% between 150 and 200 eV. PMID- 20001035 TI - Absorption and emission spectra of ultraviolet B blocking methoxy substituted cinnamates investigated using the symmetry-adapted cluster configuration interaction method. AB - The absorption and emission spectra of ultraviolet B (UVB) blocking cinnamate derivatives with five different substituted positions were investigated using the symmetry-adapted cluster configuration interaction (SAC-CI) method. This series included cis- and trans-isomers of ortho-, meta-, and para-monomethoxy substituted compounds and 2,4,5-(ortho-, meta-, para-) and 2,4,6-(ortho-, para-) trimethoxy substituted compounds. The ground and excited state geometries were obtained at the B3LYP/6-311G(d) and CIS/D95(d) levels of theory. All the compounds were stable as cis- and trans-isomers in the planar structure in both the S(0) and S(1) states, except the 2,4,6-trimethoxy substituted compound. The SAC-CI/D95(d) calculations reproduced the recently observed absorption and emission spectra satisfactorily. Three low-lying excited states were found to be relevant for the absorption in the UV blocking energy region. The calculated oscillator strengths of the trans-isomers were larger than the respective cis isomers, which is in good agreement with the experimental data. In the ortho- and meta-monomethoxy compounds, the most intense peak was assigned as the transition from next highest occupied molecular orbital (next HOMO) to lowest unoccupied molecular orbital (LUMO), whereas in the para-monomethoxy compound, it was assigned to the HOMO to LUMO transition. This feature was interpreted as being from the variation of the molecular orbitals (MOs) due to the different substituted positions, and was used to explain the behavior of the excited states of the trimethoxy compounds. The emission from the local minimum in the planar structure was calculated for the cis- and trans-isomers of the five compounds. The relaxation paths which lead to the nonradiative decay were also investigated briefly. Our SAC-CI calculations provide reliable results and a useful insight into the optical properties of these molecules, and therefore, provide a useful tool for developing UVB blocking compounds with regard to the tuning of the photoabsorption. PMID- 20001036 TI - Structure and vibrational spectra of H(+)(HF)(n) (n=2-9) clusters: An ab initio study. AB - The morphological development of the hydrogen bond network in the protonated hydrogen fluoride clusters, H(+)(HF)(n) (n=2-9), is investigated in detail by ab initio methods. We find a dominance of the linear morphology, which is energetically well separated from the other minimum energy morphologies of the clusters. The geometry for these clusters shows a pattern due to the cooperativity effect prevalent in the hydrogen bonds, as a result of the difference in electronegativities of hydrogen and fluorine atom in the HF molecule. The variations in the covalent HF and hydrogen bond distances in the clusters are in turn reflected in the vibrational spectra. Distinct HF stretching modes for the linear and ring with tail structures were identified. We have discussed the signature peaks for the two possible ion-core morphologies present in the clusters. The highly corrosive nature of HF makes it difficult to study using experiments. We, thus, believe that our structure and vibrational spectra calculations would be useful in understanding the key features in these systems. PMID- 20001037 TI - Methane molecule confined in the small and large cages of structure I clathrate hydrate: Quantum six-dimensional calculations of the coupled translation-rotation eigenstates. AB - We report fully coupled quantum six-dimensional (6D) calculations of the translation-rotation (T-R) energy levels of CH(4) molecule inside the small dodecahedral (5(12)) and large tetracaidecahedral (5(12)6(2)) cages of the structure I clathrate hydrate. The quantum dynamics of the three translational and three rotational degrees of freedom of CH(4) are treated rigorously, while the guest molecule and the host cavities are taken to be rigid. The matrix of the full 6D T-R Hamiltonian is diagonalized in the product basis of contracted translational and angular basis functions, generated by solving two reduced dimension (3D) eigenvalue problems. A pairwise additive CH(4)-cage 6D potential energy surface (PES) is employed, constructed using the anisotropic CH(4)-H(2)O pair potential which was utilized previously in the molecular dynamics simulations of methane hydrate. Our calculations elucidate the key features of the T-R energy level structure of the nanoconfined CH(4). The rotational levels of methane exhibit an elaborate pattern of splittings caused by the angular anisotropy of the environment; the splitting patterns are identical for both types of cages. Translationally excited T-R states in the small cage are assigned in terms of the quantum numbers n and l of the 3D isotropic harmonic oscillator and those in the large cage using the Cartesian quantum numbers. Extensive comparison is made with the data from the inelastic neutron scattering studies of methane hydrate, allowing an assessment of the accuracy of the 6D PES employed. PMID- 20001038 TI - The photodissociation of oxetane at 193 nm as the reverse of the Paterno-Buchi reaction. AB - We investigated the photodissociation of oxetane (1,3-trimethylene oxide) at 193.3 nm in a molecular-beam apparatus using photofragment-translational spectroscopy and selective photoionization. We measured time-of-flight (TOF) spectra and angular anisotropy parameters beta(t) as a function of flight time of products at m/z=26-30 u utilizing photoionization energies from 9.8 to 14.8 eV. The TOF distributions of the products alter greatly with the employed photon energy, whereas their beta(t) distributions are insensitive to the photon energy. Dissociation to H(2)CO+C(2)H(4) is the major channel in the title reaction. Three distinct dissociation paths with branching ratios 0.923:0.058:0.019 are responsible for the three features observed in the distribution of kinetic energy released in the channel H(2)CO+C(2)H(4). The observation of H(2) and H atoms, approximately 1% in branching, indicates that products H(2)CO and C(2)H(4) spontaneously decompose to only a small extent. Most HCO, C(2)H(3), and C(2)H(2) ions originate from dissociative photoionization of products H(2)CO and C(2)H(4). Except atomic H and H(2), the photoproducts have large angular anisotropies, beta>or=-0.8, which reflects rapid dissociation of oxetane following optical excitation at 193.3 nm. The mechanisms of dissociation of oxetane are addressed. Our results confirm the quantum-chemical calculations of Palmer et al. and provide profound insight into the Paterno-Buchi reaction. PMID- 20001039 TI - Rationalizing and functionalizing stannaspherene: Very stable stannaspherene "alloys". AB - It is illustrated here by ab initio calculations based on density functional theory and other high level methods that the high stability of the icosahedral Sn(12) (2-) dianion known as stannaspherene, reflects stability toward ionization rather than cohesion. This could be also connected with novel fluxional rearrangements and paths of Sn(12) (1-) leading eventually to Sn(12) (2-) involving charge transfer. In view of the very similar structural and electronic properties with the corresponding isovalent borane (B(12)H(12))(2-), it is demonstrated that stannaspherene can be further rationalized and functionalized on the basis of an isolobal analogy between group 14 clusters and isovalent boranes, carboranes, and bisboranes. Such analogy is of the same nature with analogous isolobal and isovalent similarities between silicon, hydrogenated silicon-carbon clusters and deltahedral boranes and carboranes, which the present author, scoptically and synoptically, has described as the "boron connection." It is predicted and verified theoretically: First, that the isovalent Bi(2)Sn(10) and Sb(2)Sn(10) clusters, considered as the microscopic analogs of tin-bismuth alloys, are very stable (more stable than stannaspherene itself) very symmetric and isolobal to Sn(12) (2-); and second, that embedded clusters of the form M@Sn(12) (2-), M@Bi(2)Sn(10), M=Pt,Pd are very stable and highly symmetrical (I(h) and D(5d) respectively) with large highest occupied-lowest unoccupied molecular orbital gaps and very large embedding energies of the order of 5-6 eV. It is furthermore predicted that Pt@Sn(12) (2-) and Pt@Bi(2)Sn(10) can be synthesized in view of their higher stability compared to Pt@Pb(12) (2-) which has already been synthesized. The marginal energy difference of 0.03 eV between the meta- and the para-isomer of Bi(2)Sn(10) indicates a fluxional behavior with respect to Bi-Sn interchange which should be related with the Sn(12) (1-) fluxionality leading eventually to Sn(12) (2-). This rearrangement is also associated with a strange aromatic behavior. The same type of Bi-Sn fluxionality is also encountered in higher energy structures. Due to the "inert pair effect" in tin, the validity of the isolobal analogy is much stronger and fully valid compared to isovalent species based on germanium or silicon, such as Ge(12) (2-), Bi(2)Ge(10), and Ge(10)C(2)H(2) and Si(12) (2-), Bi(2)Si(10), and Si(10)C(2)H(2). The present ideas are in full agreement with available experiments and suggest even further functionalization of stannaspherene, analogous to metaloboranes, metalocarboranes, and stannaboranes with several potential applications. PMID- 20001040 TI - Heavy atom nitroxyl radicals. IV. Experimental and theoretical studies of the F(2)P=S free radical in the gas phase. AB - The difluorothiophosphoryl (F(2)PS) free radical has been produced in a supersonic discharge jet from a precursor mixture of F(3)PS and high pressure argon and detected by laser-induced fluorescence and single vibronic level emission spectroscopy. With the aid of high level ab initio predictions of the properties of the ground and first two excited doublet states, the observed band system has been positively identified as B(2)A(')-X(2)A('). The electronic transition involves promotion of an electron from the pi to the pi( *) orbital with concomitant increases in the out-of-plane angle and PS bond length on excitation. The observed vibrational structure, Franck-Condon profile, rotational band contours, T(0) energy, and ground and excited state vibrational frequencies are all in accord with expectations based on our theoretical predictions. PMID- 20001041 TI - Microwave measurements and ab initio calculations of structural and electronic properties of N-Et-1,2-azaborine. AB - Rotational transitions for N-Et-1,2-azaborine were measured in the 5-13 GHz range using a Flygare-Balle type Fourier transform spectrometer system. Twelve distinct rotational transitions with over 130 resolved hyperfine components, which included a-dipole and b-dipole transitions, were measured and analyzed to obtain rotational constants and (11)B and (14)N nuclear quadrupole coupling constants in the principal rotational axis system. Rotational constants obtained are A=4477.987(4), B=1490.5083(7), and C=1230.6728(6) MHz. The quadrupole coupling constants for (11)B are eQq(aa)=-1.82(1), (eQq(bb)-eQq(cc))=-3.398(4) MHz, and for (14)N, eQq(aa)=1.25(1), (eQq(bb)-eQq(cc))=0.662(4) MHz. Quantum electronic structure calculations predict a ground-state structure with the ethyl group perpendicular to the azaborine plane and rotational constants in very good agreement with the measured structure and rotational constants. The theoretical conformational analysis of the ethyl group rotation around the N[Single Bond]C bond in relation to the heterocyclic ring yielded an asymmetric torsional potential energy surface with barrier heights of about 900 and 1350 cm(-1) for the N-Et-1,2-azaborine. Results of the measurements and calculations indicate that the basic molecular structure of N-Et-1,2-azaborine is similar to ethylbenzene. Electrostatic potential calculations qualitatively show that pi electron density is somewhat delocalized around the 1,2-azaborine ring. PMID- 20001042 TI - Intramolecular charge transfer of 4-(dimethylamino)benzonitrile probed by time resolved fluorescence and transient absorption: No evidence for two ICT states and a pisigma( *) reaction intermediate. AB - For the double exponential fluorescence decays of the locally excited (LE) and intramolecular charge transfer (ICT) states of 4-(dimethylamino)benzonitrile (DMABN) in acetonitrile (MeCN) the same times tau(1) and tau(2) are observed. This means that the reversible LE<==>ICT reaction, starting from the initially excited LE state, can be adequately described by a two state mechanism. The most important factor responsible for the sometimes experimentally observed differences in the nanosecond decay time, with tau(1)(LE)1,800 m above sea level) forests. We determined the thermal requirements for sporogonic development of P. relictum in the natural vector, Culex quinquefasciatus , and assessed the current distribution of native bird species in light of this information. Sporogonic development was completed at constant laboratory and mean field temperatures between 30 and 17 C, but development, prevalence, and intensity decreased significantly below 21 C. Using a degree-day (DD) model, we estimated a minimum threshold temperature of 12.97 C and a thermal requirement of 86.2 DD as necessary to complete development. Predicted (adiabatic lapse-rate) and observed summer threshold isotherm (13 C) correspond to the elevation of high forest refuges on the islands of Maui and Hawai'i. Our data support the hypothesis that avian malaria currently restricts the altitudinal distribution of Hawaiian honeycreeper populations and provide an ecological explanation for the absence of disease at high elevation. PMID- 20001097 TI - Seroprevalence of Trypanosoma cruzi in raccoons from Tennessee. AB - Trypanosoma cruzi is the etiologic agent of Chagas' disease. Autochthonous human and canine transmission of T. cruzi has been documented in Tennessee, but little is known about its ecology, including the prevalence of T. cruzi among wildlife in Tennessee. Serum samples from 706 raccoons (Procyon lotor) from 10 counties in the Ridge and Valley and Blue Ridge Mountains ecoregions of eastern Tennessee were tested for antibodies reactive with T. cruzi using the indirect fluorescent antibody assay. Two hundred six (29.2%) samples were seropositive, with 9 counties yielding positive samples (range 14.6-63.6%). Significantly more raccoons from rural habitats (35.1%) were found positive for T. cruzi exposure than were those from suburban habitats (23.1%, P < 0.001). Land cover class was not associated with seropositivity status (P = 0.441), even though deciduous forest was the most common site from where raccoons were trapped and the most common site of positive raccoons in rural areas (42%). Interestingly, age was positively associated with seropositivity. Raccoons older than 1 yr (adults) were 40.1% seropositive compared to 12.2% of those less than 1 yr (juveniles; P < 0.001). Female adults were significantly more likely to be exposed to T. cruzi than were male adult raccoons (P < 0.001). No significant seroprevalence difference was seen among male and female juveniles. This study contributes to understanding the dynamics of T. cruzi exposure within raccoon populations in Tennessee. The importance of habitat (rural vs. suburban) and microhabitat (dens) in risk of exposure to these populations is also discussed. PMID- 20001098 TI - Laser microdissection microscopy and single cell PCR of avian hemosporidians. AB - Avian malaria parasites (Plasmodium spp.) and related species of Haemoproteus constitute a remarkably diverse and species rich group of blood parasites. Analyses of the mitochondrial cytochrome b gene of these hemosporidians have demonstrated unexpected patterns of host distribution, host shifts, and host sharing. However, deeper insights into these patterns require access to multiple genetic markers and genetic analyses of single parasite cells. In the present study, we demonstrate the potential of laser microdissection microscopy (Olympus/MMI CellCut microdissection system) for solving these 2 problems. This technique was used for isolation of single blood stages and ookinetes of avian Haemoproteus and Plasmodium spp., which were then successfully used for DNA isolation, amplification, and sequencing. The methods of single cell dissection of hemosporidian parasites and PCR-based analyses with dissected single cells are described. These methods can be used to isolate substantial quantities of pure hemosporidian parasite DNA for large-scale sequencing, essential information when designing primers for developing multiple nuclear genetic markers. Such markers can then be applied to isolated single parasite cells for identification of parasites in mixed infections and deciphering mechanisms behind apparent reproductive isolation between parasite lineages. This method can be used in the molecular investigation of blood parasites of birds, reptiles, and fish because it enables removing the parasite DNA from the overpowering host DNA, which is present in red blood cells. PMID- 20001099 TI - Reconsidering the role of sleep for motor memory. AB - Previous studies suggest that sleep may play an important role in memory consolidation of motor skills. It has been difficult, however, to tease apart the effect of sleep from circadian and homeostatic factors. We examined the effect of sleep on a popular motor sequence task, utilizing a design that controlled for time of day and time since sleep between wake and sleep groups. When these factors were controlled, there was no benefit of sleep to motor memory, suggesting that previous work may have been influenced by circadian and homeostatic confounds. PMID- 20001100 TI - The role of the medial prefrontal cortex in the play fighting of rats. AB - Although decorticated rats are able to engage in play, their play is abnormal in three ways. First, decorticates do not display the normal, age-related shifts in defensive strategies during development. Second, decorticates do not modify their defensive tactics in response to the social identity of their partners. Third, decorticates display a global shift in defensive tactics from more complex to less complex strategies. It has been shown that lesions of the motor cortex (MC) selectively produce the abnormal developmental effects on play, and that lesions of the orbitofrontal cortex (OFC) selectively produce the deficits in behavioral discrimination between social partners. In the current set of experiments, we demonstrate that lesions of the medial prefrontal cortex (mPFC) produce the shift from more complex to less complex defensive tactics, while leaving intact the age related and partner-related modulation of defensive strategies. Thus, we have evidence for a triple dissociation of function between the MC, the OFC, and the mPFC with respect to social play behavior. PMID- 20001101 TI - GABA(A) receptor signaling in the lateral septum regulates maternal aggression in mice. AB - Maternal aggression (maternal defense) is a fierce aggression produced by lactating females toward intruders that plays an important role in protection of vulnerable offspring. Enhancement of GABA(A) receptor signaling by benzodiazepines increases maternal aggression, and we recently found indirect evidence that lateral septum (LS) could be a key site where benzodiazepines elevate aggression. In this study, we directly tested the hypothesis that activation of GABA(A) receptors in LS would promote maternal aggression while inhibition of this receptor would decrease aggression. Site-directed injections to LS were made using the GABA(A) receptor antagonist, bicuculline (3-30 ng), or the GABA(A) receptor agonists, chlordiazepoxide, a benzodiazepine (2.5-5 microg), and muscimol (0.05-5 ng). Maternal aggression and other behavioral measures were then evaluated in lactating mice. Neither GABA(A) receptor agonist elevated aggression, which could reflect a ceiling effect. However, 7 ng of the GABA(A) receptor antagonist, bicuculline, in LS significantly decreased maternal aggression without altering other maternal behaviors or light-dark box performance, suggesting some GABA(A) receptor signaling in LS is required for full maternal aggression expression. Together, these results confirm a role for GABA(A) receptor signaling in LS in the regulation of maternal aggression. PMID- 20001102 TI - A potential gastrointestinal link between enhanced postnatal maternal care and reduced anxiety-like behavior in adolescent rats. AB - Early life experience impacts emotional development in the infant. In rat pups, repeated, brief (i.e., 15 min) maternal separation (MS15) during the first 1-2 postnatal weeks has been shown to increase active maternal care and to reduce later anxiety-like behavior in the offspring. We hypothesized that the anxiolytic effect of MS15 is partly due to increased intestinal release of cholecystokinin (CCK) in rat pups as a result of increased maternal contact. We predicted that rats with a history of MS15 would display less anxiety in the elevated plus maze (EPMZ) and novelty-suppressed feeding (NSF) tests, as compared with nonseparated (NS) controls, and that the anxiolytic effect of MS15 would be attenuated in rats in which daily MS15 was accompanied by systemic administration of a CCK-1 receptor antagonist (i.e., devazepide). Treatment groups included NS control litters, litters exposed to MS15 from postnatal days (P)1-10, inclusive, and litters exposed to MS15 with concurrent subcutaneous injection of devazepide or vehicle. Litters were undisturbed after P10 and were weaned on P21. Subsets of adolescent males from each litter were tested in the EPMZ on P40-41, while others were tested for NSF on P50-52. As predicted, rats with a developmental history of MS15 displayed reduced anxiety-like behavior in the EPMZ and NSF tests. The anxiolytic effect of MS15 was preserved in vehicle-treated rats, but was reversed in devazepide-treated rats. These results support the view that endogenous CCK-1 receptor signaling in infants is a potential pathway through which maternal-pup interactions regulate the development and functional organization of emotional circuits that control anxiety-like behavior in the offspring. PMID- 20001103 TI - Interactions between the prefrontal cortex and amygdala during delay discounting and reversal. AB - Interactions between the prefrontal cortex and amygdala are thought to be critical for reward anticipation. Alterations in reward anticipation that lead to an inability to wait for rewards or a diminished capacity to change behavior when doing so would be optimal are often termed impulsivity and compulsivity, respectively. Distinct regions of the prefrontal cortex may support decreased impulsivity through self-control and decreased compulsivity through flexibility. However, both self-control and flexibility appear to involve the amygdala. Using a delay discounting paradigm, the current investigation found that inactivation and disconnection of the medial prefrontal cortex and basolateral amygdala led rats to become more impulsive by affecting preference for smaller immediate over larger delayed rewards. Conversely, inactivation and disconnection of the orbitofrontal cortex and amygdala led rats to become more compulsive as demonstrated by an inability to flexibly reverse stimulus-reward relationships in an odor reversal task. The current findings support a double dissociation between orbitofrontal cortex-amygdala interactions for odor reversal and medial prefrontal cortex-amygdala interactions for delay discounting. PMID- 20001104 TI - Transplantation of hippocampal cell lines restore spatial learning in rats with ventral subicular lesions. AB - We have demonstrated in our previous studies that ventral subicular lesion induces neurodegeneration of the hippocampus and produces cognitive impairment in rats. In the present study, the efficacy of transplanted green fluorescent protein (GFP)-labeled hippocampal cell line (H3-GFP) cells in establishing functional recovery in ventral subicular lesioned rats has been evaluated. The survival of H3-GFP transplants and their ability to express trophic factors in vivo were also investigated. Adult male Wistar rats were subjected to selective lesioning of ventral subiculum and were transplanted with H3-GFP cells into the cornu ammonis 1 (CA1) hippocampus. The transplants settled mainly in the dentate gyrus and expressed neurotrophic factors, brain-derived neurotrophic factor (BDNF), and basic fibroblast growth factor (bFGF). The ventral subicular lesioned (VSL) rats with H3-GFP transplants showed enhanced expression of BDNF in the hippocampus and performed well in eight-arm radial maze and Morris water maze tasks. The VSL rats without hippocampal transplants continued to show cognitive impairment in task learning. The present study demonstrated the H3-GFP transplants mediated recovery of cognitive functions in VSL rats. Our study supports the notion of graft meditated host regeneration and functional recovery through trophic support, although these mechanisms require further investigation. PMID- 20001105 TI - Deficit in prepulse inhibition in mice caused by dietary n-3 fatty acid deficiency. AB - Docosahexaenoic acid (DHA) and eicosapentaenoic acid (EPA) may be biosynthesized from a precursor alpha-linolenic acid (LNA) or obtained preformed in the diet. Dams were fed four diets with different levels of the various n-3 fatty acids during pregnancy and lactation, and their offspring were weaned to the same diets: "n-3 Deficient," containing (as % total fatty acids) 0.07% of LNA; "Low LNA" (0.4%); "High LNA" (4.8%); and a "DHA + EPA" diet, containing 0.4% of LNA, 2% DHA, and 2% EPA. Sensorimotor gating was measured by prepulse inhibition (PPI) of the acoustic startle response in C57Bl6 mice. The n-3 Deficient and Low LNA diets caused a substantial deficit in PPI compared to the DHA + EPA diet, whereas the High LNA diet induced a less pronounced, but significant reduction of PPI. These are the first data that demonstrate a deficit in sensorimotor gating in rodents caused by an inadequate amount of the n-3 fatty acids in the diet. Our results differentiate the effects of a High LNA diet from one with added EPA and DHA even though the difference in brain DHA content is only 12% between these dietary groups. PMID- 20001106 TI - Dissociating the conditioning and the anorectic effects of estradiol in female rats. AB - The present series of experiments challenges the ability of the hormone estradiol to act as an unconditioned stimulus in the conditioned taste avoidance (CTA) learning paradigm. We hypothesize that reductions in sucrose consumption observed after pairing it with estradiol are not indicative of associative learning, but due to the unconditioned expression of estradiol's anorectic effects during the time of CTA assessment. Three experiments in which a sucrose solution was paired with estradiol were conducted to test this hypothesis. Experiment 1 demonstrated that female rats expressed a reduction in post-pairing sucrose consumption even though the anorectic effects of estradiol had subsided. Experiment 2 showed that although a low dose of estradiol produced anorexia, it did not elicit post pairing reductions in sucrose consumption. Experiment 3 revealed that contingent pairing was a requirement for post-pairing reduction in sucrose consumption even when testing was done at a time when anorexia is expressed. These findings demonstrate the dissociability of the conditioning and anorectic effects of estradiol, providing evidence against the hypothesis. The results are discussed in terms of independent neural mechanisms underlying the disparate behaviors. PMID- 20001108 TI - Microstructural pattern of palatable food intake from weaning to adulthood in male and female OLETF rats. AB - Ontogenetic trajectories from weaning to adulthood and sex differences in feeding patterns were examined in the Otsuka Long-Evans Tokushima Fatty (OLETF) rat, an animal model of early onset overeating-induced obesity, and a natural cholecystokinin-1 receptor knockout. Overnight patterns of licking a palatable liquid diet (Ensure) were analyzed on Postnatal Days 22, 38, 60, and 90. Because different microstructure profiles may reflect alterations in the influence of positive and negative signals, we examined meal parameters to uncover developing mechanisms underlying eating behavior in this strain. OLETF rats displayed significantly greater caloric intake, larger meals (in number of licks), and more (within-meal) clusters of feeding (which were shorter in duration and contained fewer licks per cluster) than did Long-Evans Tokushima Ohtsuka (LETO) strain controls. OLETF rats also had significantly lower satiety ratios than LETO rats. Moreover, we identified sex differences in the age of emergence of microstructural patterns of obesity-related overeating, suggesting that systems other than cholecystokinin may be disrupted, possibly worsening the OLETF strain's obesity phenotype. PMID- 20001107 TI - Suppression to visual, auditory, and gustatory stimuli habituates normally in rats with excitotoxic lesions of the perirhinal cortex. AB - In 3 habituation experiments, rats with excitotoxic lesions of the perirhinal cortex were found to be indistinguishable from control rats. Two of the habituation experiments examined the habituation of suppression of responding on an appetitive, instrumental baseline. One of those experiments used stimuli selected from the visual modality (lights), the other used auditory stimuli. The third experiment examined habituation of suppression of novel-flavored water consumption. In contrast to the null results on the habituation experiments, the perirhinal lesions disrupted transfer performance on a configural, visual discrimination, indicating the behavioral effectiveness of the lesions. Implications for comparator theories of habituation are considered, and it is concluded that others' demonstrations of the sensitivity of object recognition to perirhinal cortex damage is not the result of standard habituation. PMID- 20001109 TI - MPTP lesion causes neuroinflammation and deficits in object recognition in Wistar rats. AB - Animal models of Parkinson's disease with dementia would greatly facilitate research into the underlying causes of this disorder. Here, we showed that bilateral infusion of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) into the substantia nigra pars compacta (SNc) of Wistar rats caused degeneration of nigrostriatal dopaminergic neurons, cell loss in the hippocampal CA1 area, as well as microglial activation and increase of interleukin-2 levels in several brain regions. In addition, increase of anxiety-like behavior and impairment of object recognition were observed in the MPTP-lesioned rats. These findings suggest that neuroinflammation may contribute to MPTP-induced neurodegeneration and behavioral deficits, which is suggested as an animal model of Parkinson's disease dementia. PMID- 20001110 TI - Effects of ethanol and caffeine on behavior in C57BL/6 mice in the plus-maze discriminative avoidance task. AB - INTRODUCTION: Caffeine is frequently consumed concurrent to or immediately following ethanol consumption. Identifying how caffeine and ethanol interact to modulate behavior is essential to understanding the co-use of these drugs. The plus-maze discriminative avoidance task (PMDAT) allows within-subject measurement of learning, anxiety, and locomotion. METHODS: For training, each mouse was placed in the center of the plus-maze for 5 min, and each time that the mouse entered the aversive enclosed arm, a light and white noise were turned on. At testing, each mouse was returned to the center of the maze for 3 min. No cues were turned on during testing. RESULTS: Ethanol (1.0-1.4 g/kg) dose-dependently decreased anxiety and learning, and increased locomotion. Caffeine (5.0-40.0 mg/kg) dose-dependently increased anxiety and decreased locomotion and learning. Caffeine failed to reverse ethanol-induced learning deficits. However, 1.4 g/kg ethanol blocked the anxiogenic effect of caffeine. DISCUSSION: Although caffeine and ethanol interact to modulate behavior in the PMDAT, caffeine does not reverse ethanol-induced learning deficits. Ethanol-induced anxiolysis may contribute to alcohol consumption, while ethanol's blockade of caffeine-induced anxiogenesis may contribute to co-use. PMID- 20001111 TI - Dopamine D1 and D2 antagonist effects on response likelihood and duration. AB - Experimentally induced and parkinsonian disruptions in dopamine (DA) transmission are associated with motor abnormalities that include a reduced likelihood of behavioral response initiation and an increased duration of executed responses. Here we investigated the dopamine receptor subtypes involved in regulating these two aspects of behavior. We examined the effects of D1 family (D1/D5) antagonist R(+)-7-chloro-8-hydroxy-3-methyl-1-phenyl-2,3,4,5-tetrahydro-1H-3-benzazepine hydrochloride (SCH23390; 0, 0.04, 0.08, or 0.16 mg/kg) and D2/D3 antagonist 3,5 dichloro-N-(1-ethylpyrrolidin-2-ylmethyl)-2-hydroxy-6-methoxybenzamide (+) tartrate salt (raclopride; 0, 0.2, or 0.4 mg/kg) on the likelihood and duration of a cued Pavlovian approach and a cued operant lever-press response. While the high doses of the D1 and D2 antagonists produced similar levels of overall locomotor suppression, only the D2 antagonist increased the duration of time that animals' heads remained in the food compartment during both Pavlovian and operant task performance. In contrast, D1 antagonist SCH23390 decreased the proportion of trials in which animals executed both the Pavlovian approach and operant lever press, while raclopride did not. The results suggest that D2 receptor blockade preferentially increases response duration, and, under the simple discrete-trial procedures employed here, D1 receptor blockade preferential reduces Pavlovian and operant response likelihood. PMID- 20001112 TI - Interaction effect of D4 dopamine receptor gene and serotonin transporter promoter polymorphism on the cortisol stress response. AB - Genetic variation of the serotonin transporter (SCL6A4, 5-HTT) has been associated with fear- and anxiety-related behaviors, while a polymorphism in exon III of the D4 dopamine receptor gene (DRD4) has been linked to novelty seeking. The dopaminergic and the serotonergic neurotransmitter system have been found to modulate the amygdala-connected circuitries that are crucial in emotional modulation and response to fearful stimuli. Additionally, reactivity of amygdala innervated effector systems is also essential for our understanding of anxiety related behaviors. Here, we used the stress-induced activation of the hypothalamic-pituitary-adrenal axis to investigate the impact of 5-HTTLPR and DRD4 on the cortisol stress response in 84 healthy adults. Saliva cortisol was measured during and after the Trier Social Stress Test. We found a significant main effect of DRD4: Carriers of the 7R allele exhibited lower cortisol responses. Additionally, a DRD4 by 5-HTTLPR interaction emerged: 5-HTTLPR LA/LA homozygotes showed a lower cortisol response than did S or LG allele carriers but only if they possessed at least one copy of the DRD4 7R allele. The results point to independent and joint effects of these polymorphisms on stress responsivity. PMID- 20001113 TI - Sex differences in nicotine sensitization and conditioned hyperactivity in adolescent rats neonatally treated with quinpirole: role of D2 and D3 receptor subtypes. AB - Neonatal quinpirole treatment in rats produces increased sensitivity of dopamine D2-like receptors throughout the animal's lifetime, referred to as D2 priming. There is little information on the effects of nicotine in adolescent rats, especially in a model that has clinical relevance to psychosis where increased D2 receptor sensitivity is common. Male and female rats were treated with quinpirole (1 mg/kg) or saline from postnatal (P) day 1-P21, given nicotine (0.5 mg/kg) or saline from P33 through P49, and placed into a locomotor arena for behavioral testing. Nicotine or saline treatment was preceded by the D2-like receptor antagonist eticlopride, D3 antagonist nafadotride, or saline. Conditioned hyperactivity was analyzed on P50 in the same context in a drug-free test. In females, D2 priming increased the locomotor response to acute nicotine, but did not affect subsequent nicotine sensitization, and only non-D2-primed females demonstrated conditioned hyperactivity. Eticlopride and nafadotride blocked behavioral sensitization, although nafadotride was more effective at blocking nicotine-conditioned hyperactivity in females. In males, D priming enhanced sensitization to nicotine and produced conditioned hyperactivity, which were blocked by eticlopride and nafadotride. These results have implications for psychosis and comorbidity of nicotine abuse in adolescence. PMID- 20001114 TI - Intracerebroventricular insulin improves spatial learning and memory in male Wistar rats. AB - As one of the most studied protein hormones, insulin as well as its receptor have been known to play key roles in a variety of important biological processes. Detection of insulin and its receptor in the central nervous system (CNS) has led to a rapidly growing interest in the central effects of insulin. Insulin and its receptor are located in the specific area of the CNS with a diversity of region specific functions different from its direct carbohydrate homeostasis in the periphery. The high density of insulin/insulin receptor in brain areas such as the hippocampus and cerebral cortex have shown to play an important role in higher cognitive functions, suggesting that insulin might be involved in the modulation of memory. Previous studies have offered controversial results regarding the effects of insulin on various types of memory. The aim of the present study is to determine whether intracerebroventricular (ICV) administration of insulin improves the water maze performance of rats. The experimental groups had pretraining insulin infusion (2, 4, 8, 16, and 32 mu) into the third ventricle, and then they were compared with a sham (saline) group. Insulin treatment caused an enhancing effect on spatial memory in a dose dependent manner. The low doses (2, 4, and 8 mu) of insulin had no significant effect on the water maze achievement of rats, whereas higher doses (16 and 32 mu) significantly improved the rats' performance. These results suggest that ICV administration of insulin may result in a dose-dependent improvement of memory function in rats. PMID- 20001115 TI - Progressive spatial processing deficits in a mouse model of the fragile X premutation. AB - Fragile X associated tremor/ataxia syndrome (FXTAS) is a neurodegenerative disorder that is the result of a CGG trinucleotide repeat expansion in the range of 55-200 in the 5' UTR of the FMR1 gene. To better understand the progression of this disorder, a knock-in (CGG KI) mouse was developed by substituting the mouse CGG8 trinucleotide repeat with an expanded CGG98 repeat from human origin. It has been shown that this mouse shows deficits on the water maze at 52 weeks of age. In the present study, this CGG KI mouse model of FXTAS was tested on behavioral tasks that emphasize spatial information processing. The results demonstrate that at 12 and 24 weeks of age, CGG KI mice were unable to detect a change in the distance between two objects (metric task), but showed intact detection of a transposition of the objects (topological task). At 48 weeks of age, CGG KI mice were unable to detect either change in object location. These data indicate that hippocampal-dependent impairments in spatial processing may occur prior to parietal cortex-dependent impairments in FXTAS. PMID- 20001116 TI - Transient inactivation of the ventral tegmental area selectively disrupts the expression of conditioned place preference for pup- but not cocaine-paired contexts. AB - The ventral tegmental area (VTA) plays a critical role in motivated behavior. However, it remains unclear whether intact VTA function is necessary for motivated behavior to seek contexts repeatedly paired with natural stimuli and/or pharmacological stimuli. In the present study, conditioned place preference (CPP) was induced with highly salient natural or drug stimuli attributed with strong incentive-motivational value in each of 2 female models: Postpartum females were conditioned to associate one unique context in the CPP apparatus with young offspring (pups) and a second context with a neutral stimulus, and virgin females were conditioned to associate unique contexts with cocaine (5 mg/kg ip) and saline injections. Immediately before CPP testing, each female received a microinfusion of bupivacaine bilaterally into the VTA to transiently inactivate the region; subjects were also tested after saline microinfusion into the VTA. Postpartum females' preference for the pup-paired context was abolished by VTA inactivation but was restored to high control levels after saline microinfusion. In separate tests, VTA inactivation also reduced motivated pup licking and pup retrieval in postpartum females, suggesting that intact VTA function is required for the expression of both pup CPP and motivated pup-directed behaviors. Cocaine CPP remained unaffected by VTA inactivation. Locomotion was not affected by VTA microinfusions but was severely impaired by bupivacaine microinfusions into the substantia nigra. We concluded that the VTA is differentially involved in the expression of conditioned preference for contexts paired with pups, a salient natural stimulus, and contexts paired with cocaine. PMID- 20001117 TI - The effects of aging on memory for sequentially presented objects in rats. AB - The current study investigated memory for sequentially presented objects in young rats 6 months old (n = 12) and aged rats 24 months old (n = 12). Rats were tested on a task involving three exploratory trials and one probe test. During the exploratory trials, the rat explored a set of three sequentially presented object pairs (A-A, B-B, and C-C) for 5 min per pair with a 3-min delay between each pair. Following the exploratory trials, a probe test was conducted where the rat was presented simultaneously with one object from the first exploratory trial (A) and one object from the third exploratory trial (C). Results from the exploratory trials showed no significant age-related differences in exploration, indicating that 24-month-old rats explored the object pairs as much as 6-month-old rats. The probe test demonstrated that 6-month-old rats spent significantly more time exploring object A compared to object C, indicating that young rats show intact temporal order memory for the exploratory trial objects. However, 24-month-old rats showed no preference for object A and spent a relatively equal amount of time exploring objects A and C. The results suggest that temporal order memory declines as a result of age-related changes in the rodent brain. The findings also may reflect differences in attraction to objects with different memory strengths. Since age-related differences were not detected during the exploratory trials, age-related differences on the probe trial were not due solely to decreased exploration, motivation, or locomotion. PMID- 20001119 TI - Disrupted temporal control in the R6/2 mouse model of Huntington's disease. AB - Huntington's disease is characterized by corticostriatal dysfunction and degeneration of the striatum with progressive loss of the medium spiny neurons. These circuits are important for instrumental responding, interval timing, and temporal control over motor output. We investigated the acquisition of timed operant responding in two R6/2 Huntington's Disease models, differing in CAG repeat length and genetic background (115 and 250 CAG repeats, and a mixed CBAxC57 or pure C57 background) and their corresponding wild type controls using the peak procedure. Both mouse lines exhibited similar response control deficits. In unreinforced peak trials, mice either did not learn to terminate an ongoing response past reinforcement time or required more trials to acquisition compared to the wild type mice. While transgenic and wild type mice did not exhibit differences in temporal accuracy, response curves were flatter in transgenic mice, suggesting decreased temporal control over operant responding. The results are discussed in terms of the neurobiology of interval timing, instrumental responding, and the neuropathology of HD and R6/2 mice. PMID- 20001118 TI - Examination of bilateral eyeblink conditioning in rats. AB - This experiment monitored eyelid responses bilaterally during delay eyeblink conditioning in rats. Rats were given paired or unpaired training with a tone or light conditioned stimulus (CS) and a unilateral periorbital shock unconditioned stimulus (US). Rats given paired training acquired high levels of conditioned responses (CRs), which occurred in both eyelids. However, acquisition was faster, and the overall percentage of CRs was greater in the eyelid that was ipsilateral to the US. CRs in the eyelid ipsilateral to the US also had shorter onset latencies and larger amplitudes than CRs in the contralateral eyelid. Both eyelids consistently showed high percentages of unconditioned responses (UR) to the US, and the UR amplitude decreased across training sessions in the paired group. The present study demonstrated that CRs occur robustly in both eyelids of rats given eyeblink conditioning, which is similar to previous findings in humans and monkeys. The results also showed that conditioning occurs more prominently in the eyelid that is ipsilateral to the US, which is similar to previous findings in humans, monkeys, dogs, and rabbits. PMID- 20001120 TI - Are specific emotions narrated differently? AB - Two studies test the assertion that anger, sadness, fear, pride, and happiness are typically narrated in different ways. Everyday events eliciting these 5 emotions were narrated by young women (Study 1) and 5- and 8-year-old girls (Study 2). Negative narratives were expected to engender more effort to process the event, be longer, more grammatically complex, more often have a complication section, and use more specific emotion labels than global evaluations. Narratives of Hogan's (2003) juncture emotions anger and fear were expected to focus more on action and to contain more core narrative sections of orientation, complication, and resolution than narratives of the outcome emotions sadness and happiness. Hypotheses were confirmed for adults except for syntactic complexity, whereas children showed only some of these differences. Hogan's theory that juncture emotions are restricted to the complication section was not confirmed. Finally, in adults, indirect speech was more frequent in anger narratives and internal monologue in fear narratives. It is concluded that different emotions should be studied in how they are narrated, and that narratives should be analyzed according to qualitatively different emotions. PMID- 20001122 TI - Using reappraisal to regulate unpleasant emotional episodes: goals and timing matter. AB - The hypothesis that cognitive reappraisal will have different effects on emotion as a function of regulatory goal and the timing with which reappraisals are enacted within an emotion episode was tested. Forty-one participants reappraised situations depicted in unpleasant pictures by imagining those situations getting worse (increase), staying the same (maintain), or getting better (decrease). Reappraisal instructions were delivered 2 s before (anticipatory) or 4 s after (online) picture onset. Measures of rated unpleasantness, expressive behavior (corrugator muscle activity), heart rate (HR), and electrodermal activity (EDA) were collected. Increase reappraisals produced higher rated unpleasantness, corrugator muscle activity, HR, and EDA relative to maintain reappraisals. For corrugator muscle activity and EDA, the effect of increase reappraisals was only apparent when enacted online. Decrease reappraisals produced lower rated unpleasantness relative to maintain reappraisals but had no effect on expressive behavior or autonomic physiology. The effect of decrease reappraisals did not depend on when reappraisal was enacted. These data underscore the importance of regulatory goals and the impact of regulatory timing as a moderator of emotion regulatory success within an emotion episode. PMID- 20001121 TI - Reconciling findings of emotion-induced memory enhancement and impairment of preceding items. AB - A large body of work has revealed that people remember emotionally arousing information better than neutral information. However, previous research has revealed contradictory effects of emotional events on memory for neutral events that precede or follow them: In some studies, emotionally arousing items have impaired memory for immediately preceding or following items, and in others arousing items enhanced memory for preceding items. By demonstrating both emotion induced enhancement and impairment, Experiments 1 and 2 clarified the conditions under which these effects are likely to occur. The results suggest that emotion induced enhancement is most likely to occur for neutral items that (a) precede (and so are poised to predict the onset of) emotionally arousing items, (b) have high attentional weights at encoding, and (c) are tested after a delay period of a week rather than within the same experimental session. In contrast, emotion induced impairment is most likely to occur for neutral items near the onset of emotional arousal that are overshadowed by highly activated competing items during encoding. PMID- 20001123 TI - Self-relevance processing in the human amygdala: gaze direction, facial expression, and emotion intensity. AB - How the processing of emotional expression is influenced by perceived gaze remains a debated issue. Discrepancies between previous results may stem from differences in the nature of stimuli and task characteristics. Here we used a highly controlled set of computer-generated animated faces combining dynamic emotional expressions with varying intensity, and gaze shifts either directed at or averted from the observer. We predicted that perceived self-relevance of fearful faces would be higher with averted gaze-signaling a nearby danger; whereas conversely, direct gaze would be more relevant for angry faces-signaling aggressiveness. This interaction pattern was observed behaviorally for emotion intensity ratings, and neurally for functional magnetic resonance imaging activation in amygdala, as well as fusiform and medial prefrontal cortices, but only for mild- and not high-intensity expressions. These results support an involvement of human amygdala in the appraisal of self-relevance and reveal a crucial role of expression intensity in emotion and gaze interactions. PMID- 20001124 TI - Can Duchenne smiles be feigned? New evidence on felt and false smiles. AB - We investigated the value of the Duchenne (D) smile as a spontaneous sign of felt enjoyment. Participants either smiled spontaneously in response to amusing material (spontaneous condition) or were instructed to pose a smile (deliberate condition). Similar amounts of D and non-Duchenne (ND) smiles were observed in these 2 conditions (Experiment 1). When subsets of these smiles were presented to other participants, they generally rated spontaneous and deliberate D and ND smiles differently. Moreover, they distinguished between D smiles of varying intensity within the spontaneous condition (Experiment 2). Such a differentiation was also made when seeing the upper or lower face only (Experiment 3), but was impaired for static compared with dynamic displays (Experiment 4). The predictive value of the D smile in these judgment studies was limited compared with other features such as asymmetry, apex duration, and nonpositive facial actions, and was only significant for ratings of the upper face and static displays. These findings raise doubts about the reliability and validity of the D smile and question the usefulness of facial descriptions in identifying true feelings of enjoyment. PMID- 20001125 TI - An S-shaped relationship between changes in appraisals and changes in emotions. AB - Previous research on appraisal theories of emotion has shown that emotions and appraisals are related but has not specified the nature of the relationships. This research examined the functional forms of appraisal-emotion relationships and demonstrated that for all seven appraisals studied, appraisals relate to emotions in an S-shaped (ogival) fashion: Changes in appraisals at extreme levels are associated with only small changes in emotions, but changes at moderate levels are associated with substantial changes in emotions. With a few exceptions, ogival relationships were found for the relationships between seven appraisals (Goal Achievement Expectancy, Agency, Control, Certainty, Fairness, Pleasantness, and Motive Congruence) and numerous relevant emotions across different sample-types, cultures, and methods. PMID- 20001126 TI - The voice conveys specific emotions: evidence from vocal burst displays. AB - Studies of emotion signaling inform claims about the taxonomic structure, evolutionary origins, and physiological correlates of emotions. Emotion vocalization research has tended to focus on a limited set of emotions: anger, disgust, fear, sadness, surprise, happiness, and for the voice, also tenderness. Here, we examine how well brief vocal bursts can communicate 22 different emotions: 9 negative (Study 1) and 13 positive (Study 2), and whether prototypical vocal bursts convey emotions more reliably than heterogeneous vocal bursts (Study 3). Results show that vocal bursts communicate emotions like anger, fear, and sadness, as well as seldom-studied states like awe, compassion, interest, and embarrassment. Ancillary analyses reveal family-wise patterns of vocal burst expression. Errors in classification were more common within emotion families (e.g., 'self-conscious,' 'pro-social') than between emotion families. The three studies reported highlight the voice as a rich modality for emotion display that can inform fundamental constructs about emotion. PMID- 20001127 TI - The unconscious regulation of emotion: nonconscious reappraisal goals modulate emotional reactivity. AB - People often encounter difficulty when making conscious attempts to regulate their emotions. We propose that nonconscious self-regulatory processes may be of help in these difficult circumstances because nonconscious processes are not subject to the same set of limitations as are conscious processes. Two experiments examined the effects of nonconsciously operating goals on people's emotion regulatory success. In Experiment 1, participants engaged in an anxiety eliciting task. Participants who had a reappraisal emotion control goal primed and operating nonconsciously achieved the same decrease in physiological reactivity as those explicitly instructed to reappraise. In Experiment 2, the effect of nonconscious reappraisal priming on physiological reactivity was shown to be most pronounced for those who do not habitually use reappraisal strategies. The findings highlight the potential importance of nonconscious goals for facilitating emotional control in complex real-world environments and have implications for contemporary models of emotion regulation. PMID- 20001128 TI - Fearful faces influence attentional control processes in anxious youth and adults. AB - This study examined the effects of trait anxiety and age on performance of an emotional working memory task designed to investigate attentional control processes in the context of emotion. Participants included children, adolescents, and adults (8-30 years old). They performed the Emotional Face N-Back (EFNBACK) task, a modified n-back working memory task with four emotional distracter types (no picture, neutral, fearful, and happy) and two memory-load conditions (0-back and 2-back), and completed self-report trait anxiety measures. Results indicated that participants high in trait anxiety had slower reaction times on the fearful 2-back memory-load condition. A significant interaction with age indicated that this effect was greater in the younger participants. These findings suggest that anxious individuals, particularly younger ones, exhibit difficulty resisting interference from threat-related stimuli when greater attentional resources are being recruited. PMID- 20001129 TI - Dissociating emotion-induced blindness and hypervision. AB - Previous findings suggest that emotional stimuli sometimes improve (emotion induced hypervision) and sometimes impair (emotion-induced blindness) the visual perception of subsequent neutral stimuli. We hypothesized that these differential carryover effects might be due to 2 distinct emotional influences in visual processing. On the one hand, emotional stimuli trigger a general enhancement in the efficiency of visual processing that can carry over onto other stimuli. On the other hand, emotional stimuli benefit from a stimulus-specific enhancement in later attentional processing at the expense of competing visual stimuli. We investigated whether detrimental (blindness) and beneficial (hypervision) carryover effects of emotion in perception can be dissociated within a single experimental paradigm. In 2 experiments, we manipulated the temporal competition for attention between an emotional cue word and a subsequent neutral target word by varying cue-target interstimulus interval (ISI) and cue visibility. Interestingly, emotional cues impaired target identification at short ISIs but improved target identification when competition was diminished by either increasing ISI or reducing cue visibility, suggesting that emotional significance of stimuli can improve and impair visual performance through distinct perceptual mechanisms. PMID- 20001130 TI - Emotion recognition across cultures: the influence of ethnicity on empathic accuracy and physiological linkage. AB - The present study tested whether empathic accuracy and physiological linkage during an emotion recognition task are facilitated by a cultural match between rater and target (cultural advantage model) or unaffected (cultural equivalence model). Participants were 161 college students of African American, Chinese American, European American, or Mexican American ethnicity. To assess empathic accuracy-knowing what another person is feeling-participant's (raters) used a rating dial to provide continuous, real-time ratings of the valence and intensity of emotions being experienced by 4 strangers (targets). Targets were African American, Chinese American, European American, or Mexican American women who had been videotaped having a conversation with their dating partner in a previous study and had rated their own feelings during the interaction. Empathic accuracy was defined as the similarity between ratings of the videotaped interactions obtained from raters and targets. To assess emotional empathy--feeling what another person is feeling--we examined physiological linkage (similarity between raters' and targets' physiology). Our findings for empathic accuracy supported the cultural equivalence model, while those for physiological linkage provided some support for the cultural advantage model. PMID- 20001131 TI - Memories of yesterday's emotions: does the valence of experience affect the memory-experience gap? AB - Intense pain is often exaggerated in retrospective evaluations, indicating a possible divergence between experience and memory. However, little is known regarding how people retrospectively evaluate experiences with both pleasant and unpleasant aspects. The Day Reconstruction Method (DRM; Kahneman. Krueger, Schkade, Schwarz, & Stone, 2004b) provides a unique opportunity to examine memory experience gaps in recollections of individual days, which elicit a wide gamut of emotions. We asked female participants (N = 810, Study 1, and N = 615, Study 2) to reconstruct episodes of the previous day using the DRM and demonstrated that memory and experience diverge for both pleasant and unpleasant emotions. When they rated their day overall in a retrospectively evaluative frame of mind, the participants recalled more unpleasant and pleasant emotions than they reported feeling during the individual episodes, with a larger gap for unpleasant emotions than for pleasant emotions. The findings suggest that separate processes are used for committing positive and negative events to memory and that, especially when unpleasant emotions are involved, prudence is favored over accuracy. PMID- 20001132 TI - Are moods motivational states? A study on effort-related cardiovascular response. AB - Based on the mood-behavior-model (Gendolla, 2000), this study tested the idea that moods only have effects on effort mobilization in settings that directly call for this and in which people can thus use their moods as task-relevant information. Fifty university students were randomly assigned to a 2 (Mood: negative vs. positive) x 2 (Memorizing: intentional vs. incidental) x 2 (Time: mood induction vs. task performance) mixed model design. Effort mobilization was operationalized as systolic blood pressure (SBP) reactivity. As expected, in the intentional-memorizing condition, SBP reactivity was stronger in a negative mood than in a positive mood. Mood had no impact in the incidental-memorizing condition, which did not call for effort mobilization. PMID- 20001133 TI - Basic hue-meaning associations. AB - Color may not only be pleasing to the eye, but may also carry important associations relevant for psychological functioning. Two experiments were conducted to test for basic hue-meaning associations, controlling for lightness and chroma. Specifically, we used a reaction time paradigm to test for links between red and green, and words that varied in achievement content (failure and success) or valence more generally. Results revealed that red was positively associated with failure and general negative words, and was negatively associated with success and general positive words, whereas green was positively associated with success words only. These findings directly document that hue carries psychologically relevant meaning. Implications both within and beyond the achievement domain are discussed. PMID- 20001134 TI - A closing editorial: reflections on the Journal of Family Psychology, 2004-2009. PMID- 20001135 TI - "If you can't say something nice, don't say anything at all": coping with interpersonal tensions in the parent-child relationship during adulthood. AB - Tensions are normative in the parent-child tie, but there is less information on the strategies used to cope with such tensions. This study examined strategies parents and adult children use in reaction to interpersonal tensions and the implications of those strategies for relationship quality. Parents and their adult sons and daughters (aged 22 to 49 years; N = 158 families, 474 individuals) reported the strategies they used in response to tensions with one another (constructive, destructive, and avoidant). Across dyads, parents and adult children reported using constructive strategies more often than destructive or avoidant strategies. Strategy use varied between and within dyads by generation, gender of parent, ethnicity, education, and age of child. Constructive strategies predicted better relationship quality, whereas avoidant and destructive strategies predicted poorer relationship quality. Parents may be more likely to use constructive strategies, which are meant to maintain the relationship because of their greater investment in the tie. (PsycINFO Database Record (c) 2009 APA, all rights reserved). PMID- 20001136 TI - Prenatal expectations in transition to parenthood: former infertility and family dynamic considerations. AB - Prenatal expectations are important for the future parent-child relationship. The authors examined how maternal and paternal prenatal expectations of the relationship with the child predicted 1st-year parenting stress and whether these expectations were violated over the transition to parenthood. They further examined how former infertility affected these associations. The participants were 745 Finnish couples, 367 having undergone a successful assisted reproductive treatment and 378 conceiving spontaneously. Couples completed a questionnaire of family representations during pregnancy and when the child was 2 and 12 months old and Abidin's Parenting Stress Index at 2 and 12 months postpartum. The hypothesis of moderately high expectations predicting the lowest level of parenting stress was substantiated only concerning paternal expectations of own autonomy with the child. Generally, however, negative expectations of own and spouse's relationship with the child were linearly associated with higher parenting stress. Postnatal representations were more positive or equal to expectations, except for negative violation occurring in maternal expectation of the father-child relationship, especially among normative mothers. The results are discussed in relation to family dynamic considerations and special features of formerly infertile couples. PMID- 20001137 TI - Fathering in family context and child adjustment: a longitudinal analysis. AB - This study focuses on relations between fathers' behavior in family context and children's adjustment, including the roles of paternal depressive symptoms, paternal marital conflict behaviors, paternal parenting, and children's emotional security. Participants included 235 families with a six-year-old child, with families followed longitudinally each year for three years. In terms of fathers' adjustment, paternal problem drinking was related to paternal negative marital conflict behaviors and decreased positive parenting, which was associated with children's externalizing and internalizing problems. Fathers' depressive symptoms were directly related with children's internalizing problems. Children's emotional security was an intervening variable in relations between father's behavior in family context and children's development. PMID- 20001138 TI - Opportunity for interaction? A naturalistic observation study of dual-earner families after work and school. AB - Everyday patterns of interaction can strengthen or undermine bonds between family members. This naturalistic observation study focused on an understudied facet of family life: opportunities for interaction among dual-earner family members after work and family members' responses to these opportunities. Thirty dual-earner couples and their children were observed and video-recorded in their homes throughout two weekday afternoons and evenings. Two interaction opportunities were analyzed: (1) the behavior of family members toward a parent returning home from work and (2) the physical proximity of family members throughout the evening. Three main findings emerged. Women, who tended to return home before men, were greeted with positive behavior and reports of the day's information from family members. Men, in contrast, returned home later in the day and received positive behavior or no acknowledgment from family members distracted by other activities. Throughout the evening, mothers spent more time with children whereas fathers spent more time alone. Couples were seldom together without their children. The implications of observed interaction patterns and the contribution of naturalistic observation methods to the study of family relationships are discussed. PMID- 20001139 TI - Intergenerational transmission of aggression in romantic relationships: the moderating role of attachment security. AB - This prospective study used longitudinal, multi-reporter data to examine the influence of parents' marital relationship functioning on subsequent adolescent romantic relationships. Consistent with Bryant and Conger's (2002) model for the Development of Early Adult Romantic Relationships (DEARR), we found that interactional styles, more specifically paternal aggression and satisfaction, exhibited in parents' marital relationship when their adolescents were age 13 were predictive of qualities of the adolescent's romantic relationships 5 years later. Continuities were domain specific: paternal satisfaction predicted adolescent satisfaction and paternal aggression predicted adolescent aggression. Attachment security moderated the link between paternal aggression and subsequent adolescent aggression, with continuities between negative conflictual styles across relationships reduced for secure adolescents. Results are interpreted as suggesting that attachment may help attenuated the transmission of destructive conflict strategies across generations. PMID- 20001140 TI - Beyond neighborhood poverty: family management, neighborhood disorder, and adolescents' early sexual onset. AB - The authors examined how neighborhood disorder modifies associations between family management practices and youth transitions to sex among low-income African American and Latino urban families. The sample included 846 young adolescents and their mothers who participated in Welfare, Children and Families: A Three-City Study. Results from multilevel logistic regression models indicated no main effects of family management practices or neighborhood-level conditions on transitions to sex once accounting for demographics. However, higher levels of family routines and parental knowledge (i.e., awareness of youths' friends, whereabouts, and activities) were more strongly associated with a lower probability of youth sexual onset as neighborhood disorder increased. Results provide further evidence for the contextually specific nature of parenting impacts on adolescent adjustment. PMID- 20001141 TI - Parental beliefs, infant temperament, and marital quality: associations with infant-mother and infant-father attachment. AB - The present research examined parental beliefs about the importance of the paternal caregiving role, mothers' and fathers' reports of infant temperament, and observed marital quality as predictors of infant-mother and infant-father attachment security, over and above the effects of parental sensitivity. Infants' attachment security to mothers and fathers were observed in the Strange Situation at 12 and 13 months, respectively (N = 62 two-parent families). Hierarchical regression models revealed that mothers who viewed the paternal caregiving role as important were less likely to have securely attached infants, but only when infant fussiness was high. In addition, fathers who viewed the paternal caregiving role as important were more likely to have securely attached infants, but only when infants' fussiness or marital quality was high. PMID- 20001142 TI - Marital transitions and changes in African American mothers' depressive symptoms: the buffering role of financial resources. AB - The effects of changes in marital status on the changes in depressive symptoms of 443 African American mothers from the National Longitudinal Survey of Youth 1979 (NLSY) were examined. Results showed that those mothers who exited marriage increased in depressive symptoms relative to continuously married and newly married mothers. Moreover, mothers who entered marriage later experienced the same level of depressive symptoms as continuously married mothers. However, financial resources moderated the effects of marital transitions. Those mothers with more financial resources did not experience an increase in depressive symptoms after divorce, but those with fewer resources experienced a large increase. It was concluded that divorce is a risk factor for mental health concerns among African American mothers, but financial resources serve as a protective factor. PMID- 20001143 TI - Changes in relationship satisfaction across the transition to parenthood: a meta analysis. AB - The U.S. government has recently spent several hundred million dollars to promote healthy relationships in new parents. The influx of money implies that relationships of new parents are at elevated risk for declining satisfaction and dissolution. This meta-analysis aggregates data from 37 studies that track couples from pregnancy to after the birth of the first child and 4 studies that track childless newlywed couples over time and compare couples who do and do not become parents. Results indicate significant, small declines in relationship satisfaction for both men and women from pregnancy to 11 months postbirth; 5 studies that followed couples for 12-14 months found moderate-sized declines. Seven variables moderated the decrease in relationship satisfaction from pregnancy to early parenthood. However, the decrease in satisfaction may not indicate anything unique about the transition to parenthood; the 4 studies following newlyweds indicated that those who do not become parents experience a decrease in relationship satisfaction similar to that of parents across a comparable span of time. Implications for prevention and future directions are discussed. PMID- 20001144 TI - Experiences of familial acceptance-rejection among transwomen of color. AB - Because of the stigma associated with transgenderism, many transwomen (biological males who identify as female or transgender) experience rejection or abuse at the hands of their parents and primary caregivers as children and adolescents. The Parental Acceptance-Rejection (PAR) theory indicates that a child's experience of rejection may have a significant impact on their adult lives. The purpose of this study was to conduct a qualitative analysis of adult transwomen of color's experiences with caregivers, guided by PAR theory. Twenty transwomen of color completed semi-structured interviews exploring the reaction of their parents and primary caregivers to their gender. While many participants reported that at least one parent or close family member responded with warmth and acceptance, the majority confronted hostility and aggression; reports of neglect and undifferentiated rejection were also common. Many transwomen were forced out of their homes as adolescents or chose to leave, increasing their risk of homelessness, poverty, and associated negative sequelae. Future research is needed to explore how families come to terms with having a transgender child and how best to promote acceptance of such children. PMID- 20001145 TI - Perceived parenting skill across the transition to adoptive parenthood among lesbian, gay, and heterosexual couples. AB - Little research has examined change in perceived parenting skill across the transition to parenthood or predictors of change in perceived skill. The current study used an ecological framework to examine predictors of self-perceived parenting skill among 47 lesbian, 31 gay, and 56 heterosexual couples who were adopting their first child. Findings revealed that, on average, all new parents perceived themselves as becoming more skilled, although gay men increased the most and lesbians the least. Participants who were female, reported fewer depressive symptoms, expected to do more child care, and reported higher job autonomy viewed themselves as more skilled pre-adoption. With regard to change, parents who reported more relational conflict and parents who expected to do more child care experienced lesser increases in perceived skill. These findings suggest that regardless of gender, sexual orientation, and route to parenthood, new parents experience similar, positive changes in perceived skill, thereby broadening our understanding of parenting skill in diverse groups. The findings also highlight the importance of examining how gender, sexual orientation, and the family context may shape perceived skill across the transition to parenthood. PMID- 20001146 TI - Mothers' and fathers' attributions for adolescent behavior: an examination in families of depressed, subdiagnostic, and nondepressed youth. AB - This study examined whether parents of adolescents experiencing depressive symptoms or disorder make more negative and fewer positive attributions for their adolescents' behavior than do parents of nondepressed adolescents, and whether parental attributions for adolescents' behavior contribute to parenting behavior, above and beyond the adolescents' behavior. Parents and adolescents (76 girls and 48 boys) participated in videotaped problem-solving interactions (PSIs). Each parent subsequently watched the videotape and offered attributions for their adolescent's behavior. In addition, parent and adolescent behavior during the PSIs was coded. Mothers and fathers in families of nondepressed adolescents made significantly fewer negative attributions for their children's behavior than did parents in families of adolescents with diagnostic or subdiagnostic levels of depressive symptoms. Moreover, mothers' and fathers' negative attributions were related to greater levels of observed aggressive behavior and lower levels of observed facilitative behavior during the PSIs controlling for both demographic characteristics and the relative level of adolescent aggressive and facilitative behavior during the PSI. PMID- 20001147 TI - Mother-adolescent physiological synchrony in naturalistic settings: within-family cortisol associations and moderators. AB - In this study, the authors examined parent-adolescent cortisol associations in 45 families with adolescent children (24 girls; M age = 15.78 years, SD = 1.44 years). Family members' salivary cortisol levels were measured seven times a day on 2 typical weekdays. Family members provided reports of demographic and health variables, and adolescents rated parent-child relationship characteristics. After accounting for the effects of time of day and relevant demographic and health control variables on cortisol levels, hierarchical linear models indicated the presence of significant covariation over time in mother-adolescent cortisol (i.e., physiological synchrony). Furthermore, moderating tests revealed that mother-adolescent cortisol synchrony was strengthened among dyads characterized by mothers and adolescents spending more time together, and in families rated higher on levels of parent-youth shared activities and parental monitoring or supervision. Analysis of momentary characteristics indicated that maternal presence at the time of cortisol sampling lowered adolescent cortisol levels but did not account for mother-adolescent cortisol synchrony. Within-family physiological synchrony was amplified in momentary contexts of elevated maternal negative affect and elevated adolescent negative affect. PMID- 20001148 TI - Working models of attachment to parents and partners: implications for emotional behavior between partners. AB - This study examined whether working models of attachment are associated with observed positive emotion, sadness, and anger during marital conflict. Individuals (n = 176) from a longitudinal study of families participated in the current cross-sectional study. Narrative interviews assessed the unique and combined contribution of attachment representations based on parents (adult attachment) and partner (couple attachment). The influence of partner's attachment, depression symptoms, and sex of participant was also examined. Hierarchical linear models demonstrated that one's couple attachment security predicts one's observed positive emotion, whereas the partner's couple attachment security predicts one's observed negative emotion. Partner's depression symptoms moderated the effects of partner's couple attachment. Adult attachment was not related to observed emotional behavior between partners. These findings have important clinical implications for individual, couple, and family therapy. PMID- 20001149 TI - Mother and father connectedness and involvement during early adolescence. AB - The purpose of the current study was to explore how mother's and father's connectedness and involvement individually and collectively influence the lives of their children. Specifically, we asked how fathers' and mothers' parent-child connectedness and behavioral involvement influenced both problem behaviors (externalizing and internalizing behaviors) and positive outcomes (prosocial behaviors and hope) during early adolescence. Data for this study were taken from the Flourishing Families Project, from which 349 mothers and fathers were selected, along with their early adolescent child (mean age = 11.23 years, SD = .96). Hierarchical regression analyses revealed (even after controlling for child age, gender, and self-regulation) that mothers' and fathers' contributions differed, primarily as a function of child outcome. Namely, father (but not mother) connectedness and involvement were negatively related to adolescents' internalizing and externalizing behaviors, whereas mother (but not father) connectedness and involvement were positively related to adolescents' prosocial behaviors and hope. We also found that when one parent's involvement was low (for whatever reason), the other parent's involvement made a significant and important contribution to the child's well-being, particularly in the area of internalizing behaviors. PMID- 20001150 TI - Reliability of family maltreatment diagnostic criteria: 41 site dissemination field trial. AB - R. E. Heyman and A. M. S. Slep developed and conducted an effectiveness trial of a diagnostic system for child and partner maltreatment. The current investigation tested the widespread dissemination of the diagnostic system at 41 child and partner maltreatment services sites (n = 549 partner maltreatment cases and n = 342 child maltreatment cases). Agreement between field and master reviewers' decisions was good for partner physical and emotional abuse (kappa = 0.83-0.84). Agreement was also good for child physical, emotional, and sexual abuse (kappa = 0.73-0.89) but lagged slightly for child neglect (kappa = 0.66). Thus, multifaceted and content valid family maltreatment diagnostic criteria can be disseminated reliably. Replication studies of interrater agreement of the diagnostic system in typical clinical and agency settings are necessary; however, the high levels of agreement in myriad field sites imply that consistency of maltreatment determinations is achievable in widespread use. PMID- 20001151 TI - Infection with Helicobacter pylori. Prevalence, research and impact of antibiotic resistance. PMID- 20001152 TI - Antimicrobial susceptibility of Helicobacter pylori and mechanisms of clarithromycin resistance in strains isolated from patients in Uruguay. AB - The prevalence and mechanisms of antibiotic resistance of Helicobacter pylori have not yet been investigated in Uruguay. The objective of this study was to assess the susceptibility of H. pylori to the most frequently used antibiotics and to determine the mechanism of resistance to clarithromycin. Seventy-nine isolates were obtained from gastric biopsies of 50 adult patients during two periods, 2001 and 2006. The former group enrolled the general population (GP), the latter group Afro-descendant (AD) subjects. The minimum inhibitory concentrations of clarithromycin, amoxicillin, tetracycline, metronidazole, and levofloxacin were determined using the E-test technique. Amplification was achieved through PCR and nucleic acid sequencing to detect mutations in the site of action of clarithromycin in the rRNA gene 23S. No amoxicillin or tetracycline resistant strains were found. Clarithromycin resistance was found in 12% of the patients overall: 19.4% resistance in AD patients and no resistance in the GP group. This difference was statistically significant. The highest resistance was seen with metronidazole (36%), present in similar proportions in the two groups: 36.8% (GP) and 35.5% (AD). One GP patient and one AD patient had levofloxacin resistant strains. Sequencing analysis of gene 23S rRNA showed that only mutation in position 2143 was presented in all clarithromycin-resistant strains. PMID- 20001153 TI - Decrease in viral load at weeks 12 and 24 in patients with chronic hepatitis B treated with lamivudine or adefovir predicts virological response at week 48. AB - AIM: The aim of our study was to evaluate the decrease in viral load (VL) that is able to predict antiviral treatment response at one year in patients with chronic hepatitis B. METHODS: The clinical records of 66 patients, 31 treated with lamivudine (LAM) and 35 treated with adefovir (ADF), were retrospectively reviewed. We measured viral DNA at months 1, 3 and 6. RESULTS: The LAM group showed virological response (VR) in 51.6% of patients. Baseline VL was higher in non responders (5.37 +/- 1.16 vs. 7.01 +/- 1.05; p < 0.001). Responders showed a higher percentage of VL decrease at month 3 from baseline (49.2 vs. 38.3%; p = 0.03). We designed a ROC curve and established a cutoff point for decrease of 30% that had 80% of negative predictive value (NPV).The ADF group showed VR in 57.1% of patients. Baseline VL was higher in nonresponders (4.67 +/- 1.22 vs. 5.78 +/- 1.34; p = 0.01). We observed a significant decrease in VL (log) at months 3 (2.6 +/- 1.1 vs. 1.3 +/- 1.3; p = 0.03) and 6 (2.6 +/- 1.2 vs. 1.3 +/- 1.2; p = 0.006). The percentage of decrease of VL from baseline was also statistically significant. We created ROC curves at months 3 and 6, and established the best cutoff points. At month 6 a decrease of 1 log in VL had a NPV of 80%, and a decrease of 20% in VL from baseline had 100% NPV. CONCLUSION: The decrease in viral DNA at weeks 12 and 24 can predict VR at one year in patients with chronic hepatitis B treated with LAM or ADF. This could optimize treatment. PMID- 20001154 TI - Epidemiologic study on the current incidence of inflammatory bowel disease in Madrid. AB - INTRODUCTION: The incidence of inflammatory bowel disease (IBD) varies widely according to geographical area and has been reported to have increased in the last few years. No data are available on the current incidence of this disease in Madrid (Spain). AIM: to determine the incidence of inflammatory bowel disease in the area of influence of University Hospital Fundacion Alcorcon (Madrid), and to compare our results with those from other Spanish and European series. PATIENTS AND METHODS: A prospective, population-based study was performed to determine the incidence of IBD in the area of University Hospital Fundacion Alcorcon in Madrid between 2003 and 2005. Total population: 213,587 inhabitants (177,490 older than 14 years). Crude rates and age- and sex-specific rates adjusted to the European standard population were calculated. A retrospective study (1998-2003) was also performed. RESULTS: A total of 69 cases were diagnosed -Crohn s disease (CD): 35, ulcerative colitis (UC): 33, indeterminate colitis: 1- in the prospective period. Crude rates of CD and UC were 7.92 and 7.47 cases/100,000 inhabitants/year, respectively (the population aged 0-14 years). Specific rates were 8.0 (95% CI, 7.03-8.97) and 7.47 (95% CI, 6.5-8.4), respectively. Mean age at diagnosis was 31.02+/- 10.76 and 39.91+/-16.19 years for CD and UC, respectively. Incidence in the retrospective study was 7.13 and 6.22 cases/100,000 inhabitants/year, respectively for CD and UC. CONCLUSIONS: The incidence of CD and UC in Madrid has increased in the last decades, with rates close to those in northern European countries for CD, higher than those recently published in Spanish prospective studies and similar to those previously described in Spain and southern countries for UC. Rates were higher in the prospective period than in the retrospective one. PMID- 20001155 TI - Ultrasonography is an accurate technique for the diagnosis of gastrointestinal tumors in patients without localizing symptoms. AB - INTRODUCTION AND OBJECTIVES: This study compared the accuracy of ultrasonography in the diagnosis of gastrointestinal tumors in patients with several degrees of clinical suspicion. MATERIAL AND METHODS: We recruited patients that were suspect for gastrointestinal neoplasia but with no evidence of localizing symptoms (group A), and patients that were suspect for colon cancer (group B) or for gastric cancer (group C). Accuracy in the diagnosis was compared for: gastric cancer in groups A and C, and for colon cancer in groups A and B. The comparison was made by using the contingency coefficient, which quantifies coincidence of endoscopic and ultrasonographic diagnoses. RESULTS: Seventy-nine patients were included in group A (48 males), wherein 12 colon and 5 gastric neoplasms were detected. Group B was comprised of 153 patients (78 males) and included 66 patients with colorectal cancer (CCR). Group C contained 58 patients (35 males), 31 of whom were diagnosed with gastric cancer. The accuracy of sonography for diagnosing colon cancer was 95.5% for group A and 87.5% for group B. The contingency coefficient for endoscopy vs. ultrasonography was greater for group A: 0.658 than for group B: 0.549. The diagnostic accuracy for gastric cancer was 97.4% for group A and 86.2% for group C. The contingency coefficient between endoscopic and ultrasonographic diagnoses was also greater in group A (0.618) than in group C (0.588). CONCLUSIONS: The accuracy of ultrasonography in diagnosing colon and gastric cancer is not lower in patients without localizing symptoms. PMID- 20001157 TI - Abdominal mass, multiple lymphadenopathies, and polyposis. PMID- 20001156 TI - Noninvasive assessment of liver fibrosis. Serum markers and transient elastography (FibroScan). AB - Both the prognosis and potential treatment of chronic liver disease greatly depend on the progression of liver fibrosis, which is the ultimate outcome of chronic liver damage. Historically, liver biopsy has been instrumental in adequately assessing patients with chronic liver disease. Histological assessment allows clinicians both to obtain diagnostic information and initiate adequate therapy. However, the technique is not exempt of deleterious effects. Multiple diagnostic tests have been developed for the staging of fibrosis using noninvasive methods, most of them in the setting of chronic hepatitis C. The goal of this paper is to review available data on the staging and assessment of liver fibrosis with two methods: serum markers and transient elastography (FibroScan). PMID- 20001158 TI - Presacral mass as a cause of constipation. PMID- 20001159 TI - Acquired chronic hepatocerebral degeneration due to cirrhosis from non-alcoholic steatohepatitis. AB - INTRODUCTION AND OBJECTIVE: Acquired chronic hepatocerebral degeneration, acquired hepatolenticular degeneration or pseudo-Wilson is an infrequent disorder with a hepatic origin. Cases in the literature are scarce and it is frequently confused with hepatic encephalopathy and Wilson s disease. The aim of this essay is to report a patient suffering from this disorder due to cirrhosis from non alcoholic steatohepatitis. CASE REPORT: We present a 54-year-old man diagnosed from cirrhosis grade B9 of the Child Pugh classification. He progressively developed a picture with bradylalia, mild postural and action tremor and spatial and temporal disorientation. Further studies demonstrated an increase of the values of hepatic transaminases and a hyperintensity in the basal nuclei in the cerebral magnetic resonance imaging. Clinical and radiological data established the diagnosis of hepatocerebral degeneration. CONCLUSIONS: Acquired chronic hepatocerebral degeneration is a disorder rarely reported in the literature that it is usually confused with other diseases. We alert about the need of having this diagnosis into account with patients developing neurological symptoms after hepatic disease. PMID- 20001160 TI - [Syphilitic hepatitis: case report]. PMID- 20001161 TI - [Desmoid tumor arising in a laparoscopic trocar site after cholectomy]. PMID- 20001162 TI - [Giant lipoma and intestinal intussusception]. PMID- 20001163 TI - [Intestinal obstruction by metastasis in mesentery from squamous cell lung carcinoma]. PMID- 20001164 TI - [Bowel subocclusion by Anisakis]. PMID- 20001165 TI - A rare large cutaneous ulcer from the rural area, Democratic Republic of Congo. What is the diagnosis: Buruli ulcer? PMID- 20001166 TI - Knowledge and attitudes to sun exposure among adolescents in Korinthos, Greece. AB - INTRODUCTION: Extended sun exposure may lead to serious health problems, and evidence of this is in the increase in skin cancer and malignant melanoma worldwide. Extended sun exposure during childhood or adolescence increases the probability of skin cancer in adulthood. The aim of the study was to identify and examine the knowledge, attitude, behaviour and beliefs of Greek adolescents (high school students) related to sun exposure and its adverse effects on health. The majority of participants (89.7%) were of Greek nationality. METHODS: The study took place in 5 schools in the prefecture of Korinthos, and 816 of the total 925 students aged 15-18 years participated. The questionnaire was pilot tested and assessed for validity and reliability, both of which were adequate (Cronbach's alpha = 0.70 and r(s) = 0.78); SPSS 13.0 software was used for analysis. RESULTS: Only 37.9% of participants knew that melanoma was a type of skin cancer; 50% said they did not know what melanoma was. Regarding behavior, 35.5% reported that during the last summer they went to the beach on 20 to 50 occasions, and only 50% reported that they wore a sunhat or stayed in the shade. The frequency of sunscreen use was alarmingly low, with the majority of the adolescents unfamiliar with its proper use, and 50% not using a sunscreen with sufficient sun protection factor. Television was an important source of information about protection from sun exposure, while the family was the most important provider of advice. CONCLUSIONS: Participants' knowledge of sun exposure was insufficient and they reported risky behaviours in the summer months. Despite health promotion and community education programs focusing on sun smart strategies, these young people still associated suntans with beauty. Health promotion and education programs need to challenge such beliefs. However, as a sole approach to health promotion, teaching protective measures and appropriate ways for youth to protect themselves against the harmful effects of sunbathing may be insufficient to reduce the epidemic of skin cancer. PMID- 20001167 TI - Variation in outpatient antibiotic prescribing in the United States. AB - OBJECTIVE: To evaluate variation in outpatient antibiotic utilization among US commercial health plans and the implications of this variation for cost and quality. STUDY DESIGN AND METHODS: We measured antibiotic utilization rates among 229 US commercial health plans that participated in the 2005 Healthcare Effectiveness Data and Information Set. Rates were adjusted to account for health plan age and sex distribution. To estimate antibiotic costs, we multiplied utilization data for each drug class by national estimates of intraclass distribution of drugs, duration of therapy, and median average wholesale price. RESULTS: Antibiotic utilization rates varied markedly among plans, ranging from 0.64 antibiotic fills per member per year (PMPY) at the 5th percentile of plans to 1.08 fills PMPY at the 95th percentile, with a mean of 0.88 (SD +/- 0.15) antibiotic fills PMPY. US census region was the strongest predictor of antibiotic utilization. Antibiotic costs averaged $49 PMPY and ranged from $34 to $63 PMPY among plans at the 5th and 95th percentiles of cost, respectively. If a health plan with 250,000 members at the 90th percentile of antibiotic costs reduced its costs to the 25th percentile, annual drug cost savings would be approximately $4.1 million. CONCLUSIONS: Antibiotic utilization varies substantially among commercial health plans and is not accounted for by differences in the age and sex distribution of plan members. Because reducing rates of antibiotic utilization is likely to lower costs and improve quality, high-utilizing plans may reap considerable rewards from investing in programs to reduce the overuse of antibiotics. PMID- 20001168 TI - Effect of antihypertensive medication adherence among employees with hypertension. AB - OBJECTIVES: To determine if antihypertensive medication adherence is associated with decreased medical and drug costs, medical service utilization, and work absence days. STUDY DESIGN: Retrospective database study using medical, pharmacy, sick leave, short-term and long-term disability, and workers' compensation claims data from multiple large US employers from 2001 to 2008. METHODS: We used medical and pharmacy claims to identify employees with hypertension. The index date was the date of the first hypertension-related pharmacy claim. Eligible employees had health plan enrollment 6 months before the index date and at least 12 months after the index date. Employees younger than 45 years were excluded from the study. Regression models estimated the effect of the proportion of days covered (PDC) by hypertension medication on outcomes after the index date, including health benefit costs, medical service utilization, and work absence days, as well as some clinical outcomes calculated separately for high-prior-cost and low-prior cost employees. High-prior-cost employees were those who accounted for the top 60.0% of total medical costs during the 6 months before the index date. The regression models controlled for demographics, job-related variables, and comorbidities. RESULTS: Among low-prior-cost employees, high PDC was associated with increased medical and drug costs and work absence days. Among high-prior cost employees, high PDC was associated with decreased medical and drug costs, fewer work absence days and inpatient hospital days, and increased hypertension specific medical costs. CONCLUSION: Antihypertensive medication adherence was associated with improvement in some short-term utilization measures among high prior-cost employees, but significant short-term improvement was not seen among low-prior-cost employees. PMID- 20001169 TI - Plan designs that encourage the use of generic drugs over brand-name drugs: an analysis of a free generic benefit. AB - OBJECTIVE: To evaluate the effect on adherence and medical care expenditures of a pharmacy benefit change that included free generic drugs and higher copayments for brand-name drugs. STUDY DESIGN: Quasi-experimental pre-post study of patients with ischemic heart disease (1286 control and 555 intervention) and patients with diabetes mellitus (4089 control and 1846 intervention). METHODS: Medical and pharmacy claims data were analyzed for continuously enrolled members from January 1, 2005, through December 31, 2008. A generalized linear model was used to predict costs as adherence changed. RESULTS: The rate of switching from brand name drugs to generic drugs in the intervention group was not statistically different from that in the control group. The net change in adherence was higher only for the intervention group patients taking statins who switched to generic drugs, a 6.2% increase compared with an 8.5% decrease in the control group. The estimate of medical cost savings attributable to this benefit change was significant for only the metformin class of diabetes drugs. Improved adherence independent of this benefit change was estimated to reduce all-cause medical costs for patients taking sulfonylureas, metformin, and thiazolidinediones. CONCLUSIONS: Altering copayments for pharmaceuticals may affect the rate of conversion to generic drugs but is unlikely in and of itself to result in complete conversion. However, increasing adherence can result in net savings for specific diabetic drug classes, as savings from all-cause medical costs offset the increase in pharmacy costs. PMID- 20001170 TI - Can managed care plans reliably infer the quality of cardiac surgeons' outcomes? AB - OBJECTIVE: To assess whether managed care plans can reliably infer the quality of cardiac surgeons' outcomes. STUDY DESIGN: Evaluation of administrative discharge data and reported health plan enrollments. METHODS: We analyzed 221,327 coronary artery bypass graft (CABG) admissions performed by 398 cardiac surgeons in 75 state-regulated hospitals in Florida between 1998 and 2006. For our outcomes quality measure, we constructed surgeon-level risk-adjusted mortality rates using demographic and comorbidity data. We also obtained managed care plan enrollments in Florida in 2005 to discern the number of patient outcomes possibly seen by any individual plan. Finally, we constructed a confidence interval around any particular surgeon's CABG outcomes quality and tested whether the surgeon's quality could reliably be found to be worse than benchmarks using normal approximations and exact binomial limits. RESULTS: Even if a plan had as high as a 50% share of a county's managed care-insured CABG patients, then of 86 surgeons in the 5 largest counties in Florida, no surgeon could confidently be judged to be of poorer than average quality. CONCLUSIONS: In cardiac surgeons' outcomes quality monitoring, individual managed care plans face a "law of small numbers." Insufficient patient volume by contracted surgeons, inadequate variation in outcomes, and low levels of adverse outcomes combine to make true quality almost impossible to infer. Some mitigation may be possible through more effective use of data (more measures and pooling over time) and through more effective interorganizational sharing of data (leveraging specialist society quality data and statewide pooling). PMID- 20001171 TI - Analytic models to identify patients at risk for prescription opioid abuse. AB - OBJECTIVE: To assess the feasibility of using medical and prescription drug claims data to develop models that identify patients at risk for prescription opioid abuse or misuse. STUDY DESIGN: Deidentified prescription drug and medical claims for approximately 632,000 privately insured patients in Maine from 2005 to 2006 were used. Patients receiving prescription opioids were divided into 2 mutually exclusive groups, namely, prescription opioid abusers and nonabusers. METHODS: Potential risk factors for prescription opioid abuse were incorporated into logistic models to identify their effects on the probability that a prescription opioid user was diagnosed as having prescription opioid abuse. Different models were based on data available to prescription monitoring programs and managed care organizations. Best-fitting models were identified based on statistical significance (P 95%ile-for-age and-sex BMI) versus nonoverweight (<95%ile BMI) children did not differ in their beliefs about ideal child body size. CONCLUSION: Latino parents of overweight children did not differ from parents of nonoverweight children with respect to their KAB about childhood obesity. PMID- 20001193 TI - Partner communication and HIV testing among US Chinese college students. AB - OBJECTIVES: To explore the role of partner communication and relationship status on HIV testing among Chinese/Chinese American college students in northern California. METHODS: We used a cross-sectional study design to conduct focus groups and an online or paper-pencil questionnaire (N = 230). RESULTS: The majority (60%) of respondents were in committed partnerships, with 21% in casual partnerships. Less than one third (30%) of respondents had ever obtained an HIV test. History of HIV tested was associated with relationship status and partner communication. CONCLUSIONS: Interventions to increase partner communication skills while taking into account sociocultural factors among this population and their partners are needed. PMID- 20001192 TI - Subjective life expectancy and health behaviors among STD clinic patients. AB - OBJECTIVES: To examine whether people who expect to live longer engage in healthier behaviors relative to people who expect to live shorter lives. METHODS: Patients (994 black Americans, 373 white Americans) from a publicly funded clinic estimated their expected survival age and reported their health behaviors (alcohol, tobacco, and drug use; sexual behaviors; physical activity; and sleep). RESULTS: Subjective life expectancy (SLE) was a significant predictor of health behavior for both men and women. Race moderated the SLE-health behavior relation but only for alcohol and tobacco use. CONCLUSIONS: Future research should explore the potential health benefits of shifting SLE from a more limited to an expansive perspective. PMID- 20001194 TI - Social self-control, sensation seeking and substance use in samples of US and Russian adolescents. AB - OBJECTIVE: To compare the relations of social self-control and sensation seeking with substance use across samples of US and Russian adolescents. METHODS: Cross sectional data were obtained from 362 tenth-graders from Ufa, Russia, and 965 tenth-graders from California. RESULTS: Lack of social self-control was significantly related with higher alcohol and hard drug use in the Russian sample and higher cigarette use in the US sample. Higher sensation-seeking showed significant associations with higher cigarette and alcohol use in the Russian sample and higher alcohol, marijuana, and hard drug use in the US sample. CONCLUSION: As with US adolescents, prevention programs for Russian adolescents may also benefit from being tailored to higher sensation-seekers and including self-control skills training. PMID- 20001195 TI - Multimodal psychoanalytically informed aid work with children traumatized by the Chechen war. AB - As demonstrated in three cases, this paper illustrates how psychoanalytically informed multimodal care was an essential element of effective medical treatment of children traumatized by the Chechen war. Multimodal psychoanalytically informed aid work involves holding a variety of psychoanalytic viewpoints, including but not restricted to those represented by the Freudian, Interpersonal, and Relational orientations; its purpose is to allow for greater clarity in conceptualizing the traumatized child's response to war in order to provide the necessary care during the therapeutic process. Among the issues addressed are how traumatic memory can initially be expressed nonverbally, and therefore the use of embodied life-metaphors and witnessing are central to the survivors' ability to remember and symbolize. In addition, the significance of cultural awareness and sensitivity are explored as key components to the children's care. In the first case, the author illustrates how a traumatic life-metaphor can be resolved at an embodied, rather than an exclusively verbal, level. In the second case, cultural tradition and relativism have a significant impact on addressing medical and quality of life issues for the child. The third case illustrates how the analyst functions as recognizing witness to a parent's trauma; the "being with" of the relationship becomes the agent of the parent's change. PMID- 20001196 TI - Guntrip's concept of the regressed ego. AB - This paper explores what is perhaps the most important contribution of Harry Guntrip to psychoanalysis: namely his concept of the "regressed ego." In the course of his psychoanalytic work, he found certain concepts to be problematic. Therefore, while he valued some of Freud's ideas, he challenged his emphasis upon the importance of the instincts. Guntrip then drew extensively on the more object relational approach of Fairbairn and Winnicott. Nevertheless, although Guntrip believed that all these figures described important dimensions of psychological experience, he felt that they failed to acknowledge that layer which he called the "regressed ego." In his analysis of the "regressed ego," Guntrip made a valuable contribution to psychoanalytic theory and practice. However, because he thought that with this idea he had discovered the underlying cause of most psychological disturbances, he engaged in a form of reductionism and therefore undermined the value of his observations about this layer of psychological experience. PMID- 20001197 TI - A psychoanalytic study of Edward de Vere's The Tempest. AB - There is now abundant evidence that Freud was correct in believing Edward de Vere (1550-1604) wrote under the pseudonym "William Shakespeare." One common reaction is "What difference does it make?" I address that question by examining many significant connections between de Vere's life and The Tempest. Such studies promise to bring our understanding of Shakespeare's works back into line with our usual psychoanalytic approach to literature, which examines how a great writer's imagination weaves a new creation out of the threads of his or her life experiences. One source of the intense controversy about de Vere's authorship is our idealization of the traditional author, about whom we know so little that, as Freud noted, we can imagine his personality was as fine as his works. PMID- 20001198 TI - Psychoanalysis and qualitative psychotherapy research-some epistemological remarks. AB - The author discusses the extent to which psychoanalytic treatments can be tested by qualitative research strategies. He shows that these strategies operate without an adequate and epistemological, i.e., theoretically justified, methodology and that the specificity of psychoanalytic treatments in this kind of research has so far eluded us. The author suggests that the course of the treatments should be systematized relative to their outcomes within the theory of psychoanalytic treatment. Provided that the treatment theory is conceptually based on "consensual common ground" (Wallerstein, 1998, p. 1041), such studies allow us to conclude that psychoanalytic treatments will be successful if the sequences that are generalized in the treatment theory are actually realized in treatments that recognize the patients' unique requirements. PMID- 20001199 TI - Evaluation of psychodynamic psychotherapy in a community mental health center. AB - This study describes an evaluation of the effectiveness of psychodynamic psychotherapy provided in an outpatient community mental health clinic. The study used a single group pretest-posttest design involving 78 clients. Clinical outcomes included overall psychosocial functioning and quality of life, level of subjective distress, interpersonal functioning and role functioning, measured by the Outcome Questionnaire (Lambert, Hansen, Umpress, Lunnen Okilshi, & Burlingame, 2000). Clients showed statistically significant improvement from pretest (first sessions) to completion of treatment in overall functioning, and quality of life, level of subjective distress, interpersonal functioning and role functioning. Eighty-five percent of clients made statistically and clinically significant change. Calculation of effect sizes for each outcome found moderate to strong change effects ranging from d = .4 to .9. The study illustrates a method of intervention research that therapists and agencies can use to integrate practical evaluation methods into their clinical services in order to improve mental health service to clients, to demonstrate the effectiveness of interventions, and to provide data to support coverage for needed services for clients. PMID- 20001200 TI - A view from Riggs: treatment resistance and patient authority-XII. Examined living: a psychodynamic treatment system. AB - Hospitalized patients with treatment resistant comorbid Axis I and II psychiatric disorders have in common a developmental history that includes a lack of fit between their needs and their human environment. Adaptation of their personalities to this environmental failure contributes to these patients' lack of trust in authority. Treatment resistance may represent a repetition of this early experience, with the current biological and symptom focus of the field representing yet another insufficient environment. The author describes the components of a psychodynamic treatment institution that focuses on overcoming this dilemma through studying the process of relationships, attending to meaning, and maximizing patient authority. These principles may be applicable in a range of treatment settings where clinical staff members work with so-called "treatment resistant" patients. PMID- 20001201 TI - Series epilogue: a view from Riggs: treatment resistance and patient authority. PMID- 20001203 TI - FRONTLINE-the turn away from social activism in psychoanalysis. PMID- 20001202 TI - Mirror neurons, the self, and culture: an essay in neo-psychoanalysis. PMID- 20001204 TI - Hepatocyte growth factor ameliorates progression of interstitial injuries in tubular epithelial cells. AB - OBJECTIVE: Hepatocyte growth factor (HGF) and its c-met receptor comprise a signalling system that has been reported to prevent injury in several models of renal disease; however, whether HGF can also retard progression of chronic kidney disease is not known. The aim of the present study was to examine the effects of HGF on progression of chronic kidney disease in tubular epithelial cells. MATERIAL AND METHODS: Studies were performed in human tubular epithelial cells that underwent different glucose concentrations, and then receive HGF or vehicle. The cell apoptosis was tested by DAPI and TUNEL staining. The level of activity of HGF was examined at multiple time-points. The expression of HGF, transforming growth factor-beta(1) (TGF-beta(1)), plasminogen activator inhibitor-1 (PAI-1) and collagen IV were examined by reverse transcription-polymerase chain reaction, Western blot and enzyme-linked immunosorbent assay. RESULTS: HGF administration was associated with a reduction in TGF-beta(1). The beneficial effects of HGF were associated with reductions in the expression of TGF-beta(1), and in the extent of epithelial cell apoptosis. HGF appeared to induce fibrinolytic pathways by reducing the expression of collagen IV and decreasing levels of PAI-1. CONCLUSIONS: These findings suggest that HGF can retard progression of diabetic nephropathy, primarily by promoting matrix degradation, and that HGF is a potent antifibrogenic factor. PMID- 20001205 TI - Enhanced TLR4 endothelial cell immunohistochemical expression in symptomatic carotid atherosclerotic plaques. AB - BACKGROUND AND PURPOSE: Toll-like receptor-4 (TLR4) has been linked to the pathogenesis of atherosclerosis. Carotid atheroma endothelial cells (ECs) express TLR4, nevertheless correlations with cerebrovascular symptomatology, epidemiological and clinical variables remain unresolved. METHODS: Carotid atherosclerotic plaques were obtained by standard carotid endarterectomy from 157 patients with carotid artery disease (84 asymptomatic - Group A, 73 symptomatic - Group B). TLR4 expression was detected by immunohistochemistry and TLR4 positivity, overexpression and intensity of immunostaining in ECs were correlated with cerebrovascular symptomatology, epidemiological and clinical variables. RESULTS: A significant association was found between TLR4 positivity in ECs and the occurrence of any cerebrovascular event (overall response (OR): 2.85, 95% CI 1.33 - 6.11, p = 0.009). TLR4 overexpression and staining intensity in ECs were both significantly enhanced in symptomatic patients (p < 0.0001 and p = 0.003, respectively). These associations were stronger for the occurrence of a major cerebrovascular accident (CVA) compared with a transient ischemic attack (TIA) or amaurosis fugax. TLR4 expression in ECs was less prominent in statin users (OR: 0.25, 95%CI 0.1 - 0.58, p = 0.001], while it was enhanced in restenotic plaques compared with primary atherosclerotic lesions (p = 0.012). CONCLUSIONS: TLR4 expression in ECs of carotid atheroma was enhanced in symptomatic patients with most commonly 'unstable' - 'more prone to rupture' carotid plaques. PMID- 20001206 TI - Modulation of the renin-angiotensin-aldosterone system in sepsis: a new therapeutic approach? AB - IMPORTANCE OF THE FIELD: Severe sepsis is characterized by relative hypotension associated with a high cardiac output, peripheral vasodilation, and organ dysfunction. The renin-angiotensin-aldosterone system (RAAS) is primarily activated to increase blood pressure, but recently potential pro-inflammatory effects of angiotensin II have attracted interest because of the reported association between angiotensin II levels and organ failure and mortality in sepsis. RAAS antagonists could represent a new therapeutic option in this setting. AREAS COVERED IN THIS REVIEW: The role of RAAS activation in severe sepsis and septic shock, and the potential benefits (and risks) of using RAAS antagonists. WHAT THE READER WILL GAIN: Insight into RAAS function in severe sepsis and the potential for RAAS inhibitors to be used as an adjunctive therapy in patients with severe sepsis, with discussion of promising results from animal models of sepsis. TAKE HOME MESSAGE: Use of RAAS antagonists is an emerging therapeutic option in severe sepsis because these agents may reduce endothelial damage, organ failure, and mortality. However, timing of administration of RAAS antagonists is important because reduced RAAS function may contribute to refractive hypotension later on in septic shock and benefits of RAAS antagonists seem to be restricted to the early phases of sepsis. PMID- 20001207 TI - Targeting the TNF-alpha pathway in sarcoidosis. AB - IMPORTANCE OF THE FIELD/SIGNIFICANCE: Sarcoidosis is a granulomatous disease with various organ manifestations in which TNF-alpha has been demonstrated to play a major pathogenic role. The existing conventional therapies are not always able to minimize TNF-alpha-driven inflammation and other approaches should be used. AREAS COVERED IN THIS REVIEW: TNF-alpha roles in sarcoid inflammation and granuloma formation are reviewed based on the literature published in the last two decades and the therapies able to target it specifically or non-specifically in sarcoidosis are discussed. WHAT THE READER WILL GAIN: A better understanding of the pathogenic role of TNF-alpha in sarcoidosis and of the scientific rationale of its therapeutic blockade. TAKE HOME MESSAGE: In some subsets of sarcoidosis with more rapid progession and/or therapeutic refractoriness TNF-alpha plays a more prominent role in disease pathogenesis, and its blockade might represent an appropriate therapeutic approach. PMID- 20001208 TI - Targeting CD44 in mast cell regulation. AB - IMPORTANCE OF THE FIELD: Accumulating evidence suggests that mast cells are involved in a wide variety of immune responses including chronic inflammation, immune tolerance and tumor immunity. Mast cells originate from hematopoietic stem cells and undergo terminal differentiation in the tissues, in which they are ultimately resident. Heterogeneity of tissue mast cells is, therefore, one of the key concepts for a better understanding of various immune responses. AREAS COVERED IN THIS REVIEW: This review describes the candidate genes involved in regulation of cutaneous mast cell differentiation, with a particular attention to CD44, which is the primary receptor for hyaluronan. WHAT THE READER WILL GAIN: CD44 is involved in various aspects of cutaneous inflammation. Regarding mast cells, CD44 is upregulated upon differentiation and maturation of mast cells, and plays a critical role in regulation of cutaneous mast cell number. Since both degradation and decrease of hyaluronan are often observed upon chronic inflammation, CD44 might be involved in modulation of local immune responses through regulation of cutaneous mast cell functions. TAKE HOME MESSAGE: Understanding of cutaneous immune responses should require clarification of local mast cell functions, a part of which is regulated by extracellular matrix components and their membrane receptors. PMID- 20001210 TI - Targeting AAC-11 in cancer therapy. AB - IMPORTANCE OF THE FIELD: Since its discovery in 1997, the antiapoptotic factor AAC-11 has rapidly gained attention due to its potential use in cancer therapy. Indeed, most cancer cells express elevated levels of AAC-11, which is now known to be involved in both tumor cells growth as well as sensitivity to chemotherapeutic drugs. AREAS COVERED IN THIS REVIEW: In this review, we examine the most recent evidence about the role of AAC-11 in cancer biology and the therapeutic perspectives associated with its specific targeting. For that purpose, literature dealing with AAC-11 in the PubMed database was reviewed from 1997 up to date. WHAT THE READER WILL GAIN: AAC-11 is an antiapoptotic gene that has the potential to be a target for anti-cancer therapy, and warrants further investigation. As its expression seems to predict unfavorable prognosis, at least in some cancers, it also may become a potent prognostic marker. TAKE HOME MESSAGE: Blocking AAC-11 function in cancer for therapeutic purposes might be of great interest. The recent report of efficient AAC-11 inhibiting peptides that sensitize tumor cells to chemotherapeutic drugs has raise the exciting notion that AAC-11 might be a druggable target and fueled the search for new therapeutic agents that could block AAC-11 function. PMID- 20001211 TI - RhoGDI2 as a therapeutic target in cancer. AB - IMPORTANCE OF THE FIELD: Rho GDP dissociation inhibitor 2 (RhoGDI2) has been identified as a regulator of Rho GTPases that play important roles in the development of numerous aspects of the malignant phenotype, including cell cycle progression, resistance to apoptotic stimuli, neovascularization, tumor cell motility, invasiveness, and metastasis. Although RhoGDI2 has been known to be expressed only in hematopoietic tissues, recent studies suggest that this protein is also aberrantly expressed in several human cancers and contributes to aggressive phenotypes, such as invasion and metastasis. Hence, RhoGDI2 appears to be a target of interest for therapeutic manipulation. AREAS COVERED IN THIS REVIEW: Here, we summarize the role of RhoGDI2 in human cancers, specifically metastasis-related processes, and discuss its potential as a therapeutic target. WHAT THE READER WILL GAIN: RhoGDI2 modulates the invasiveness and metastatic ability of cancer cells through regulation of Rac1 activity. TAKE HOME MESSAGE: RhoGDI2 may be a useful marker for tumor progression in human cancers, and interruption of the RhoGDI2-mediated cancer cell invasion and metastasis by an interfacial inhibitor may be a powerful therapeutic approach to cancer. PMID- 20001209 TI - The NF-kappaB activation pathways, emerging molecular targets for cancer prevention and therapy. AB - IMPORTANCE OF THE FIELD: Nuclear factor kappa B (NF-kappaB) is activated by a variety of cancer-promoting agents. The reciprocal activation between NF-kappaB and inflammatory cytokines makes NF-kappaB important for inflammation-associated cancer development. Both the constitutive and anticancer therapeutic-induced NF kappaB activation blunts the anticancer activities of the therapy. Elucidating the roles of NF-kappaB in cancer facilitates developing approaches for cancer prevention and therapy. AREAS COVERED IN THIS REVIEW: By searching PubMed, we summarize the progress of studies on NF-kappaB in carcinogenesis and cancer cells' drug resistance in recent 10 years. WHAT THE READER WILL GAIN: The mechanisms by which NF-kappaB activation pathways are activated; the roles and mechanisms of NF-kappaB in cell survival and proliferation, and in carcinogenesis and cancer cells' response to therapy; recent development of NF-kappaB-modulating means and their application in cancer prevention and therapy. TAKE HOME MESSAGE: NF-kappaB is involved in cancer development, modulating NF-kappaB activation pathways has important implications in cancer prevention and therapy. Due to the complexity of NF-kappaB roles in different cancers, careful evaluation of NF kappaB's in each cancer type is crucial in this regard. More cancer cell-specific NF-kappaB inhibiting means are desired for improving anticancer efficacy and reducing systemic toxicity. PMID- 20001212 TI - Targeting focal adhesion kinase signaling in tumor growth and metastasis. AB - IMPORTANCE OF THE FIELD: Focal adhesion kinase (FAK), a crucial mediator of integrin and growth factor signaling, is a novel and promising target in cancer therapy. FAK resides within focal adhesions which are contact points between extracellular matrix (ECM) and cytoskeleton, and increased expression of the kinase has been linked with cancer cell migration, proliferation and survival. The aim of this review is to summarize the current research in the area and to assess the potential of different FAK-targeting strategies for cancer therapy. AREAS COVERED IN THIS REVIEW: We briefly examine the evidence pointing towards FAK as potential anti-cancer target since its discovery in 1992. Then, we summarize different approaches developed to interfere with FAK signaling and important results reported from these experiments. Finally, we discuss the potential of these strategies to accomplish inhibition of tumor growth and distant spread as well as potentially meaningful combinations with other therapeutic modalities in the context of the currently available evidence. WHAT THE READER WILL GAIN: The review emphasizes the link between FAK biology and the consequences of interference with FAK signaling. Based on this foundation an opinion is formed with regard to the future of FAK as therapeutic target. TAKE HOME MESSAGE: Inhibition of FAK harbours the potential to restrain malignant growth and progression with minimal side effects in normal tissues. Small molecule inhibitors of the kinase should be examined in further clinical studies and combinations with existing therapies need to be explored. More efforts are required to identify markers which predict response towards FAK inhibition. PMID- 20001215 TI - Bolus delivery of mesenchymal stem cells to injured vasculature in the rabbit carotid artery produces a dysfunctional endothelium. AB - Endothelial dysfunction is an important factor in cardiovascular pathology. It has been suggested that pluripotent mesenchymal stem cells (MSCs) may contribute to repair of the endothelium through paracrine pathways. Enhanced re endothelialization may be associated with a better outcome following angioplasty procedures. We examined the effect of the delivery of MSCs to a denuded vessel in vivo. The right carotid arteries of New Zealand white rabbits were denuded using an uninflated 3-French Fogarty balloon catheter. 1 x 10(5) MSCs in a bolus of 150 microL were then delivered intraluminally and allowed to dwell for 20 min. MSC engraftment was assessed using PKH-26 labeling and transduction with adenoviral reporter genes. Vessels were examined at 2 weeks for levels of endothelialization, as well as for neointimal hyperplasia and vasomotor function. Engraftment of MSCs was noted in the vessel wall following local arterial delivery. Endothelialization was improved following bolus MSC delivery at 2 weeks post-intervention. However, this endothelium is manifestly dysfunctional as indicated by a significant impairment in vasomotor activity and a significant increase in neointimal formation post-bolus delivery. Consistent with the formation of a dysfunctional endothelium, there was a higher rate of vessel occlusions in bolus-treated vessels due to not only predominately thrombosis but also neointimal hyperplasia. Our results suggest that naive MSCs delivered as a bolus to the occluded injured vascular segment generate dysfunctional endothelium presenting a risk of vessel occlusion. Such risks are important and need to be further assessed. PMID- 20001214 TI - Novel discoveries in understanding the complexities of epilepsy and major depression. AB - The 39th Annual meeting of the Society for Neuroscience was held in Chicago, Illinois, USA from 17 to 21 October, 2009. The conference was attended by more than 33,000 delegates from across the globe including scientists from both basic and clinical settings. Co-incidentally, this year, the scientific community is commemorating the 200th anniversary of the birth of the famous English naturalist and biologist, Charles Darwin, who described the theory of natural selection. Keeping its traditions, the congress discussed various new advances in the area of neuroscience. The topics were divided into symposia, mini-symposia, nano symposia, special lectures and poster sessions. The main areas of discussion were novel discoveries in Alzheimer's, Parkinson's, drug addiction, autism, epilepsy and major depression. According to the WHO, neurological disorders are one of the greatest threats to public health. There are many unknown and challenging facts in the field of neuroscience that needs exploration. It is unfortunate that despite the availability of various drugs for treating these disorders, a sizeable population still do not achieve complete remission. Therefore, organizing such events and addressing the latest developments may open new treatment vistas for patients suffering from these disorders. The present review discusses some of the outcomes of the deliberations in the field of epilepsy and major depression. PMID- 20001216 TI - Acute intravenous injection toxicity study of MIBG in mice. AB - Metaiodobenzylguanidine (MIBG) is an analog of norepinephrine. I-131-labeled MIBG has been thought to be safe and effective in the evaluation of neuroendocrine tumors, mainly in neuroblastoma and pheochromocytoma. This article describes the acute toxicity of MIBG in imprinting control region (ICR) mice. Treated mice were administered with MIBG at dose levels of 75, 150, and 300ng/kg with dose volumes of 20mL/kg. The control mice were administered 20mL/kg of vehicle control. The mice were observed for 14 days. Observations included general demeanor, clinical signs, mortality, body weights/total body-weight gains, and gross necropsy findings. None of the animals died during the 14-day study period. There was no difference in body weights among all treated and control mice. PMID- 20001213 TI - Targeting Pyk2 for therapeutic intervention. AB - IMPORTANCE OF THE FIELD: The focal adhesion tyrosine kinases FAK and Pyk2 are uniquely situated to act as critical mediators for the activation of signaling pathways that regulate cell migration, proliferation and survival. By coordinating adhesion and cytoskeletal dynamics with survival and growth signaling, FAK and Pyk2 represent molecular therapeutic targets in cancer as malignant cells often exhibit defects in these processes. AREAS COVERED IN THIS REVIEW: This review examines the structure and function of the focal adhesion kinase Pyk2 and intends to provide a rationale for the employment of modulating strategies that include both catalytic and extra-catalytic approaches that have been developed in the last 3 - 5 years. WHAT THE READER WILL GAIN: Targeting tyrosine kinases in oncology has focused on the ATP binding pocket as means to inhibit catalytic activity and downregulate pathways involved in tumor invasion. This review discusses the available catalytic inhibitors and compares them to the alternative approach of targeting protein-protein interactions that regulate kinase activity. TAKE HOME MESSAGE: Development of specific catalytic inhibitors of the focal adhesion kinases has improved but significant challenges remain. Thus, approaches that inhibit the effector function of Pyk2 by targeting regulatory modules can increase specificity and will be a welcome asset to the therapeutic arena. PMID- 20001217 TI - Short communication: case report on adulterated opium-induced severe lead toxicity. PMID- 20001218 TI - Metabonomic profiling of diet-induced hyperlipidaemia in a rat model. AB - This study describes the metabolic profiles of the development of hyperlipidaemia in a rat model, utilizing metabonomics by gas chromatography-mass spectrometry (GC-MS) determination coupled with multivariate statistical analysis. Rat plasma samples were collected before and during a high-lipid diet at days 0, 7, 14, 21 and 28, and were analysed for lipid levels using kit assays or metabonomics using GC-MS. Forty-one endogenous metabolites were separated, identified and quantified using GC-MS. The data matrix was processed by principal component analysis or partial least squares discriminant analysis. Dynamic modification of the rat metabonome can be clearly identified and tracked at different stages of hyperlipidaemia in the rat model. Potential biomarkers, including beta hydroxybutyrate, tyrosine and creatinine, were identified. The current work suggests that metabonomics is able to provide an overview of biochemical profiles of disease progress in animal models. Using a metabonomic approach to identify physiopathological states promises to establish a new methodology for the early diagnosis of human diseases. PMID- 20001219 TI - In vitro evaluation of a Folate-bovine serum albumin-doxorubicin conjugate. AB - Doxorubicin (DOX) is one of the most effective anticancer drugs. However, its therapeutic effectiveness is greatly hampered by its dose limiting and cumulative cardiotoxic side effects. To overcome these limitations, bioconjugates of DOX were studied using bovine serum albumin (BSA) as a carrier to provide passive tumor targeting by the enhanced permeability and retention (EPR) effect. Folic acid, as an active targeting agent, was linked to BSA to increase the selectivity of the conjugate. In the present study, folate-targeted (Folate-BSA-DOX) conjugates were prepared. In the optimization process, we found that 30 mg of folic acid activated esters reacted with BSA at pH 9.8 for 1 h, the yield was maximum. The qualitative analysis of fluorescent experiments revealed that Folate BSA-DOX can be specifically delivered to Hela cells and that this unique interaction can be blocked by 1 mM free folic acid. More importantly, the enhanced efficiency of uptake of Folate-BSA-DOX by Hela cells was coupled with the increase of the amount of the conjugate, the incubated time and the conjugated ratio of folic acid. Finally, the quantitative data obtained from the flow cytometry further verified the higher targeting and killing ability of Folate-BSA-DOX to folate receptor positive tumor cells than BSA-DOX. PMID- 20001220 TI - The inductive effect of bone morphogenetic protein-4 on chondral-lineage differentiation and in situ cartilage repair. AB - OBJECTIVES: As recent studies have suggested that bone morphogenetic protein-4 (BMP-4) and BMP-7 are promising cartilage differentiation factors, this study aimed to compare the efficacy of BMP-4 and BMP-7 for chondral-lineage differentiation in vitro as well as the efficacy of BMP-4 for articular cartilage repair in vivo. METHODS: Rabbit mesenchymal stromal cells and articular chondrocytes were treated with 10 ng/mL human recombinant BMP-4 or BMP-7. The expression of cartilage-specific genes (col II, aggrecan, and Sox9) and fibroblast growth factor receptor genes was tested by real-time polymerase chain reaction in vitro. Also, full-thickness cartilage defects (diameter 4 mm, thickness 3 mm) were created in New Zealand white rabbits and untreated (group I), or treated with a bilayer collagen scaffold (group II) or BMP-4 with scaffold (group III) (n = 12/group). The repaired tissues were harvested for histology and mechanical testing after 6 or 12 weeks. RESULTS: Cartilage differentiation of mesenchymal stromal cells was more apparent after BMP-4 treatment, as evidenced by higher expression of type II collagen and aggrecan genes. Also, BMP-4 induced higher aggrecan and fibroblast growth factor receptor-2 gene expression in chondrocytes, whereas BMP-7 had no effect. In the in vivo experiments, group III treated with BMP-4 protein had the largest amounts of cartilage tissue, which restored a greater surface area of the defect and achieved higher International Cartilage Repair Society scores. Moreover, Young's modulus, which indicates the mechanical properties of the repaired tissue, was markedly higher in group III than in groups I and II (p < 0.05), but lower than in normal tissue. CONCLUSION: BMP-4 is more potent than BMP-7 for cartilage differentiation. The delivery of BMP-4 protein in a bilayer collagen scaffold stimulates the formation of cartilage tissue. PMID- 20001221 TI - Different responsiveness of endothelial cells to vascular endothelial growth factor and basic fibroblast growth factor added to culture media under gravity and simulated microgravity. AB - When incubated under simulated microgravity (s-microg), endothelial cells (EC) form tubular structures that resemble vascular intimas. This delayed formation of 3D EC structures begins between the 5th and 7th day of culturing EC under conditions of s-microg, when double-row cell assemblies become visible. With the aim of learning about this initial phase of tubular structure formation, we found that NFkappaBp65 protein content was similar in all cell populations, but gene and protein expression of phosphokinase A catalytic subunit, phosphokinase Calpha, and extracellular signal-regulated kinases 1 and 2 was altered in cells cultured under s-microg. Apoptosis remained below 30% in all EC cultures. In contrast to controls, the 7-day-old s-microg cultures contained 3D aggregates with proliferating cells, enhanced numbers of necrotic cells, and osteopontin negative EC as well as supernatants with reduced quantities of vascular endothelial growth factor (VEGF), basic fibroblast growth factor (bFGF), soluble TNFRSF5, TNFSF5, intercellular adhesion molecule-1, tumor necrosis factor receptor 2, IL-18, complement C3, and von Willebrand factor. VEGF and/or bFGF (10 ng/mL) application influenced the accumulation of proteins in supernatants more profoundly under 1 g than under s-microg. These findings provide evidence that phosphokinase Calpha plays a key role in tube formation. Improving the interaction of VEGF and/or bFGF with EC under s-microg could enhance the engineering of vascular intimas. PMID- 20001222 TI - The relationship between primary health care organization and quality of diabetes care. AB - BACKGROUND: Despite many quality improvement trials, diabetes care often remains suboptimal. Few studies in a primary care setting have investigated the 'real life' association between organizational differences and quality of diabetes care. METHODS: Observational study among ten health care centres with a total of 45 general practitioners (GP). We investigated health care organization and related this to quality of care in a total of 1849 electronic patient records. RESULTS: There were large differences among health care centres in the percentage of patients receiving optimal care (range: 8-67%). The odds to receive good quality of care was higher if the health care centre had a diabetes education program (OR: 4.3; CI: 3.4-5.4), when yearly medical check-ups were done by both the GP and nurse practitioner (NP) (OR: 5.5; CI: 4.2-7.3), planned that after the patient visited the NP the patient is discussed with the GP (OR: 1.8; CI: 1.6 2.0), and had structured follow-up measures for compliance to check-ups (OR: 0.7; CI: 0.5-0.9 and OR: 0.59; CI: 0.5-0.7 for respectively one and two active measures compared to three active measures). CONCLUSION: Also in real life, quality of care for type 2 diabetic patients is related to health care organization. PMID- 20001223 TI - Monobactam and aminoglycoside combination therapy against metallo-beta-lactamase producing multidrug-resistant Pseudomonas aeruginosa screened using a 'break point checkerboard plate'. AB - Metallo-beta-lactamase-producing multidrug-resistant Pseudomonas aeruginosa (MDR P. aeruginosa) is a cause of life-threatening infections. With parenteral colistin not available in Japan, we treated MDR P. aeruginosa sepsis with monobactam and aminoglycoside combination therapy, with screening using a 'break point checkerboard plate'. PMID- 20001224 TI - Evaluation of glycopeptide prescription and therapeutic drug monitoring at a university hospital. AB - The aim of this study was to assess the appropriateness of glycopeptide prescription almost 15 y after the publication of the Hospital Infection Control Practices Advisory Committee (HICPAC) guidelines. We also assessed the adequacy of dose regimen and therapeutic drug monitoring (TDM). All glycopeptide prescriptions were collected during a 3-month prospective study and evaluated by 2 independent infectious diseases experts. Appropriateness of prescription was assessed according to local guidelines based on the HICPAC recommendations. A total of 154 prescriptions were evaluated: 77% (69.1-83.0) were appropriate and 36% (28.2-43.8) were adequate with regard to dose regimen and loading dose. Multivariate analysis showed greater appropriateness for vancomycin than for teicoplanin (p=0.01). There was a wide discrepancy among units (p=0.04). TDM was appropriately performed in 40% (32.3-47.7) of glycopeptide treatments. When required, dose regimen adaptations occurred in 58% of cases. In conclusion, we show a satisfactory appropriateness of glycopeptide prescription. However, the adequacy of dose regimens must be improved. Finally, TDM does not comply with recent recommendations in most cases. PMID- 20001225 TI - Adenosine deaminase and tuberculous meningitis--a systematic review with meta analysis. AB - Tuberculous meningitis (TBM) is a severe infection of the central nervous system, particularly in developing countries. Prompt diagnosis and treatment are necessary to decrease the high rates of disability and death associated with TBM. The diagnosis is often time and labour intensive; thus, a simple, accurate and rapid diagnostic test is needed. The adenosine deaminase (ADA) activity test is a rapid test that has been used for the diagnosis of the pleural, peritoneal and pericardial forms of tuberculosis. However, the usefulness of ADA in TBM is uncertain. The aim of this study was to evaluate ADA as a diagnostic test for TBM in a systematic review. A systematic search was performed of the medical literature (MEDLINE, LILACS, Web of Science and EMBASE). The ADA values from TBM cases and controls (diagnosed with other types of meningitis) were necessary to calculate the sensitivity and specificity. Out of a total of 522 studies, 13 were included in the meta-analysis (380 patients with TBM). The sensitivity, specificity and diagnostic odds ratios (DOR) were calculated based on arbitrary ADA cut-off values from 1 to 10 U/l. ADA values from 1 to 4 U/l (sensitivity >93% and specificity <80%) helped to exclude TBM; values between 4 and 8 U/l were insufficient to confirm or exclude the diagnosis of TBM (p = 0.07), and values >8 U/l (sensitivity <59% and specificity >96%) improved the diagnosis of TBM (p < 0.001). None of the cut-off values could be used to discriminate between TBM and bacterial meningitis. In conclusion, ADA cannot distinguish between bacterial meningitis and TBM, but using ranges of ADA values could be important to improve TBM diagnosis, particularly after bacterial meningitis has been ruled out. The different methods used to measure ADA and the heterogeneity of data do not allow standardization of this test as a routine. PMID- 20001226 TI - VIM-2 beta-lactamase in Pseudomonas aeruginosa isolates from Zagreb, Croatia. AB - The aim of this investigation was to characterize metallo-beta-lactamases (MBLs) in Pseudomonas aeruginosa isolates from Zagreb, Croatia. One hundred P. aeruginosa isolates with reduced susceptibility to either imipenem or meropenem were tested for the production of MBLs by MBL-Etest. The susceptibility to a wide range of antibiotics was determined by broth microdilution method. The presence of bla(MBL) genes was detected by polymerase chain reaction (PCR). Hydrolysis of 0.1 mM imipenem by crude enzyme preparations of beta-lactamases was monitored by UV spectrophotometer. Outer membrane proteins were prepared and analysed by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). Six out of 100 isolates were positive for MBLs by Etest. All strains were resistant to gentamicin, ceftazidime and cefotaxime, and all except 1 were resistant to imipenem. Six strains positive for MBLs by Etest were identified as VIM MBL producers by PCR. Sequencing of bla(VIM) genes revealed the production of VIM-2 beta-lactamase in all 6 strains. This investigation proved the occurrence of VIM 2 beta-lactamase among P. aeruginosa strains from Zagreb, Croatia. VIM-2 beta lactamase with similar properties has previously been described in another region of Croatia and in Italy, France, Spain, Greece, Taiwan and South Korea, suggesting that this type of enzyme is widespread in the Mediterranean region of Europe and in the Far East. PMID- 20001227 TI - Isolation and characterization of multipotent rat tendon-derived stem cells. AB - Stem cells have recently been isolated from humans and mice but not from rat tendon tissue. This study reports the isolation and characterization of stem cells from rat tendon. Nucleated cells isolated from rat flexor tendon tissues after collagenase digestion were plated at a low cell density to allow the selective proliferation of tendon-derived stem cells. About 1-2% of the cells isolated under this optimized culturing condition showed clonogenicity, high proliferative potential at low seeding density, and osteogenic, chondrogenic, and adipogenic multidifferentiation potential. These cells were CD44(+), CD90(+), CD34(-), and CD31(-). Although they shared some common properties with mesenchymal stem cells, they also exhibited their unique characteristics by expressing tenogenic and chondrogenic markers. There was expression of tenogenic markers, including alpha-smooth muscle actin, tenascin C, and tenomodulin, but not collagen type I at passage 0 (P0) and P3. Expression of a chondrogenic marker, aggrecan, was observed at P0 and P3, whereas expression of collagen type II was observed in few cells only at P3. The successful isolation of tendon derived stem cells under the optimized growth and differentiation conditions was useful for future stem-cell-based tissue regenerative studies as well as studies on their roles in tendon physiology, healing, and disorders using the rat model. PMID- 20001228 TI - De-staining and re-staining mucins in formalin fixed paraffin sections. AB - Re-staining of formalin fixed paraffin sections sometimes is required and this requires prior de-staining. Some simple and effective protocols for de-staining are described. Mucihematoxylin and mucicarmine can be removed with acid alcohol. Zirconyl hematoxylin can be removed with periodic acid or Sinha's fixative. Alcian blue can be removed with 5% trifluoroacetic acid in dichloromethane. Colloidal iron can be bleached in 1% household bleach in alcohol. PAS can be removed with hydrogen peroxide or ammonium hydroxide. With few exceptions, de stained sections can be re-stained with mucihematoxylin, PAS or Gabe's trichrome. PMID- 20001229 TI - Cytotoxic effects of novel phytosphingosine derivatives, including N,N dimethylphytosphingosine and N-monomethylphytosphingosine, in human leukemia cell line HL60. AB - Novel phytosphingosine derivatives have been developed based on the inhibition of sphingosine kinase, which has been implicated in cell growth and inhibition of ceramide-mediated apoptosis. This study evaluated the cytotoxic effects and underlying mechanisms of action of novel phytosphingosine derivatives, including N-monomethylphytosphingosine (MMPH) and N,N-dimethylphytosphingosine (DMPH) and the pegylated forms MMPH-PEG and DMPH-PEG, in human leukemia HL60 cells. In viability and proliferation assays using WST-1, all four drugs induced suppression of cell growth and viability in a concentration-dependent manner. Among them, DMPH had the highest antileukemic activity and induced apoptosis via caspase-8, caspase-3, and caspase-9 activation. The apoptotic effect was also associated with Fas/FasL upregulation, Bid cleavage, Bcl-2 downregulation, Bax upregulation, mitochondrial membrane depolarization, and cytochrome c release. DMPH decreased the phosphorylation of ERK and inhibited daunorubicin-induced ERK activation. Furthermore, DMPH displayed synergistic cytotoxicity with daunorubicin in a sequence-dependent manner. Our findings indicate that DMPH has potential as an effective cytotoxic agent for leukemia. PMID- 20001230 TI - Genetic changes including gene copy number alterations and their relation to prognosis in childhood acute myeloid leukemia. AB - We studied a series of 68 subjects diagnosed with childhood acute myeloid leukemia (AML) using conventional cytogenetics and fluorescence in situ hybridization (FISH), polymerase chain reaction (PCR) to analyze mutations in FLT3 and NPM1 genes, and/or array comparative genomic hybridization (CGH). Cytogenetic/FISH abnormalities were observed in 71% of subjects, FLT3-ITD mutations in 15%, and NPM1 mutations in 13%. The array CGH alterations (average 3.6 per case) were observed in 96% of the tested subjects. The most frequent alterations were gains of 8q24.3 and 11p15.5-p15.4 in 16% of the samples. Six genes (AKT1, RUNX1, LTB, SDC1, RUNX1T1, and JAK2) from the imbalanced regions have been reported to be involved in AML, whereas other 30 cancer genes, not previously reported in an AML context, were identified as imbalanced. They probably correspond to non passenger alterations that cooperate with the recurrent translocations. The clinical data and genetic changes were tested to find out the possible association with prognosis. Genomic instability (four or more genomic imbalances) was correlated with poor patient outcome (p = 0.029). PMID- 20001231 TI - Vascular endothelial growth factor concentrations from platelets correlate with tumor angiogenesis and grading in a spontaneous canine non-Hodgkin lymphoma model. AB - Published data strongly suggest that tumor progression and malignancy are associated with increased angiogenesis. However, no data have been published concerning the relationship between microvascular density (MVD), tumor cytosol, and blood vascular endothelial growth factor (VEGF) concentrations in canine non Hodgkin lymphoma (C-NHL), a neoplasm that shares biological and clinical characteristics with human NHL. We have evaluated MVD and tumor cytosol, serum (S), platelet-poor plasma (P-PP), plasma-activated platelet rich (P-APR) VEGF concentration in a series of 63 B-cells C-NHL by means of immunohistochemistry and enzyme-linked immuno-sorbent assay (ELISA) detection of VEGF. We found that MVD, VEGF from cytosol, and VEGF from P-APR are significantly correlated (p ranging from 0.001 to 0.003) and that these parameters paralleled with the malignancy degree of NHL. Accordingly, spontaneous C-NHL seems to be an interesting model to study the role of angiogenesis as interspecies pathway of tumor malignancy and we suggest that VEGF from P-APR might be a novel useful circulating bio-marker of tumor angiogenesis. PMID- 20001232 TI - Current treatment options for adult patients with Philadelphia chromosome positive acute lymphoblastic leukemia. AB - The clinical management of patients with Philadelphia chromosome-positive (Ph+) acute lymphoblastic leukemia (ALL) has been challenging primarily due to the aggressive nature of the disease and limited effective treatment options. The outcome for patients who receive conventional chemotherapy alone is poor, with remission duration of around 12 months and disease-free survival (DFS) rates of not more than 10%. Allogeneic stem cell transplantation (alloSCT) has been the only known curative treatment option, but is limited by the availability of a matched donor and the risk of treatment-related mortality. Given the role of BCR ABL in the leukemogenesis of Ph+ ALL, current treatments have focused on inhibition of this oncogenic tyrosine kinase. Early studies demonstrate that the use of the BCR-ABL tyrosine kinase inhibitor (TKI), imatinib, before alloSCT results in improved response rates and DFS when combined with standard chemotherapy regimens. Remission duration also is improved when combination chemotherapy and imatinib are administered intensively, even in the absence of allogeneic stem cell transplant. However, resistant disease remains an important problem, and the mechanisms underlying resistance in Ph+ ALL are multifactorial. Novel TKIs are currently under development and are effective in some patients with imatinib-resistant disease. The dual BCR-ABL/SRC family kinase inhibitor, dasatinib, has shown promising activity in the treatment of Ph+ ALL after imatinib failure and has recently been approved in this indication. Other TKI based therapies are also showing potential in imatinib-resistant disease. This article reviews current and emerging treatments in Ph+ ALL. PMID- 20001233 TI - Lymphoma cells contribute to the augmentation of plasma sL-selectins in the serum of lymphoma-bearing mice. AB - Like many integral membrane glycoproteins, the extracellular domain of L-selectin undergoes rapid shedding, which occurs on both resting and activated host leucocytes. Incubating normal or transformed leukocytes with phorbol esters can also artificially induce shedding of L-selectin, providing multiple possibilities for the source of soluble forms of L-selectin found in the serum of patients with hematological malignancies. Here, using genetically engineered L-selectin deficient mouse models, we have measured the release of soluble circulating forms of L-selectin in the serum of lymphoma-bearing mice. We found that L-selectin deficient lymphoma cells could not induce an elevation of circulating soluble forms of L-selectin in normal mice, as compared to lymphoma cells expressing L selectin. Moreover, soluble forms of L-selectin were detected in the serum in mice bearing lymphoma induced by injection of T lymphoma cells expressing L selectins. Interestingly, we also found that lymphoma cells that are unable to shed L-selectin in vitro following exposure to phorbol ester can generate soluble forms of serum L-selectin in vivo. Taken together, these results indicate that lymphoma cells are the major contributors to levels of soluble forms of L selectins in lymphoma-bearing mice. PMID- 20001234 TI - Low-dose fludarabine increases rituximab cytotoxicity in B-CLL cells by triggering caspases activation in vitro. AB - Rituximab maintenance therapy provides a significant benefit in patients with indolent B-cell non-Hodgkin lymphoma (NHL). Based on its efficacy in improving response to chemotherapy, the anti-CD20 antibody is currently under evaluation as maintenance therapy also in patients with B-CLL. We evaluated rituximab-mediated cytotoxicity in 10 B-CLL cases pretreated in vitro with non-cytotoxic concentrations of fludarabine. This combination induced a synergic cytotoxic effect in 8 out of 10 patients at a mean level of 26.15 +/- 13.9%, compared to 8.05 +/- 5.3% cytotoxicity observed with rituximab alone. Consistent with the viability assay, we found an increased caspase-3 activity together with activation of caspase-9 in B-CLL cells sensitive to sequential non-cytotoxic fludarabine and rituximab exposure. Non-cytotoxic fludarabine concentrations may sensitize B-CLL cells to rituximab-mediated cytotoxicity via caspase-3 activation. PMID- 20001235 TI - More (on) prognostic factors in chronic lymphocytic leukemia. PMID- 20001236 TI - Spontaneous apoptosis and proliferation detected by BCL-2 and CD71 proteins are important progression indicators within ZAP-70 negative chronic lymphocytic leukemia. AB - In chronic lymphocytic leukemia (CLL), inhibition of spontaneous apoptosis determines a worse prognosis and increasing evidences show that disease progression relies also upon cycling CLL cells. We investigated bcl-2, as measure of apoptosis, and CD71, as measure of proliferation, by flow cytometry in 265 patients with CLL. Combining bcl-2 with CD71 values, we defined three subgroups: (1) bcl2 - CD71-; (2) bcl2 + CD71+; and (3) bcl2 + CD71- or bcl2- CD71+. Both a shorter progression-free survival (PFS) and overall survival (OS) were observed in ZAP-70+ (p < 0.00001) and in patients with bcl2 + CD71+ (p < 0.00001 and p = 0.02). The patients with discordant in bcl2 + CD71- and bcl2- CD71+ showed an intermediate outcome. Noteworthy, patients with bcl2 + CD71+ showed a shorter PFS within ZAP-70 negative subgroup (p = 0.00009). In multivariate analysis of PFS, age (p = 0.005), beta-(2) microglobulin (B(2)-M) (p = 0.003), bcl-2 (p = 0.004), CD49d (p = 0.001), and ZAP-70 (p < 0.001) resulted to be significant prognostic factors. The independent prognostic significance of B(2)-M (p = 0.009) and bcl-2 (p = 0.03) was confirmed within ZAP-70 negative patients. Bcl-2 and CD71 can be considered as interesting progression indicators, which should be validated in an independent cohort of patients, to take timely therapeutic decisions in CLL. PMID- 20001237 TI - Alternaria sinusitis in children with acute myeloid leukemia: case reports from the Childhood Acute Myeloid Leukemia Infection Research Group. PMID- 20001239 TI - Hematogones: a review and update. AB - Our knowledge regarding the nature and function of 'hematogones' has evolved considerably, since the initial description more than 70 years ago. Once considered the 'mystery cells' of the bone marrow, major advances in immunology and flow cytometry have enabled us to better characterize these cells and recognize them as physiologic precursors of B-cells. In this review, we describe the morphologic and phenotypic characteristics and clinical relevance of hematogones, and report recent advances in our understanding and knowledge of these cells as they relate to physiologic and different pathologic conditions. PMID- 20001238 TI - Prognostic significance of acute renal injury in acute tumor lysis syndrome. AB - Acute tumor lysis syndrome (ATLS) caused by the destruction of malignant cells leads to metabolic abnormalities, which may either remain isolated (biological ATLS) or subsequently lead to renal dysfunction (clinical ATLS). We compared hospital and 6-month survival in patients with ATLS with hematological malignancies with or without acute renal injury. Sixty-three patients (median age, 50 years; range, 32-64) were included with ATLS. Twenty-eight had no ARI (including 17 (61%) who subsequently required dialysis) whereas 35 had an ATLS related ARI (including 31 (89%) who required dialysis). Acute leukemia (n = 28) and lymphoma (n = 30) were the main malignancies. All patients had high tumor burdens. Hospital and 6-month mortality rates were significantly lower in patients without ARI (7% and 21%, respectively) than in the ATLS-related renal injury group (51% and 66%). After adjustment for acute disease severity, presence of ARI at ICU admission was associated with higher hospital mortality (odds ratio, 10.41; 95% confidence interval, 2.01-19.170; p = 0.005) and 6-month mortality (odds ratio, 5.61; 95% confidence interval, 1.64-54.66; p = 0.006), compared to patients without renal injury. Our study suggests that in patients with ATLS, ICU management when acute renal injury is present is associated with higher short- and long-term mortality. PMID- 20001240 TI - Multifocal polyclonal Epstein-Barr virus-associated B-cell lymphoproliferative disorder secondary to azathioprine therapy successfully treated with rituximab. PMID- 20001241 TI - RUNX1 amplification in B-cell acute lymphoblastic leukemia. PMID- 20001242 TI - Bortezomib plus dexamethasone can improve stem cell collection and overcome the need for additional chemotherapy before autologous transplant in patients with myeloma. AB - The aim of this phase II trial was to investigate the efficacy of bortezomib plus dexamethasone (Vel-Dex) as induction therapy in patients with multiple myeloma (MM) and to define the role of intensification before transplantation. Fifty seven patients were treated with four courses of Vel-Dex, two cycles of dexamethasone, cyclophosphamide, etoposide and cisplatin (DCEP), and a single autologous transplant. Fourteen patients (25%) went off-study: seven after Vel Dex, seven after DCEP. All patients yielded high numbers of stem cells (median CD34+ cells 7.5 x 106/kg); 54 of the 57 patients (94%) collected > or =4 x 106/kg CD34+ cells, 60% with a single leukapheresis. The overall response rate (ORR) after Vel-Dex was 86% (70% had a very good partial response [VGPR] or better) regardless of cytogenetic abnormalities and International Staging System stage (ISS). The response at the end of the two DCEP cycles remained unchanged in 35 patients (70%), worsened in 15 (20%), and improved in 5 (10%). Because of the consistent drop-out, the ORR in intention-to-treat analysis decreased significantly from 86% after Vel-Dex to 76% after DCEP, and 73% after transplantation. However, when considering the subset of 43 patients who completed the program, the ORR was 96% (complete response 39%, VGPR 41%, partial response 16%). In conclusion, Vel-Dex produces high response rates, improves stem cell collection, and overcomes the need for intensification before autologous transplantation. PMID- 20001243 TI - Last exit clofarabine? PMID- 20001244 TI - p38 in chronic myelogenous leukemia: a target and a possible biomarker. PMID- 20001245 TI - Interim fluorodeoxyglucose positron emission tomography for early response assessment in diffuse large B cell lymphoma: where are we now? AB - Fluorodeoxyglucose (FDG)-positron emission tomography (PET) has been established in response assessment after treatment of diffuse large B cell lymphoma (DLBCL) and has been incorporated in the International Workshop Criteria (IWC). Early response assessment is increasingly being used in trials and clinical practice with the hope that it has a greater power to discriminate between good and poor prognosis patients at an earlier point in time. Early data have been very promising and suggest that PET after two to three cycles predicts progression free survival with high accuracy. Early prediction of prognosis has the potential of enabling early treatment tailoring according to the individual prognosis. Studies are yet to be done to examine the effect of changing therapy early on the basis of poor metabolic response. However, the prognosis of responders/non responders has been different in various studies, with few recent studies casting some doubt on the value of interim PET. Some of these differences are due to variability in patient populations and treatment protocols. However, there are also important factors related to the methodology of PET scanning and the criteria used to assess response. There is a need for a consensus on a uniform methodology for the use of interim PET both in clinical trials and routine clinical practice. This article critically reviews the current status and suggests future questions for further research. PMID- 20001246 TI - A novel Janus Kinase 2 mutation involving 14th intron splicing region in a patient with essential thrombocythemia. PMID- 20001249 TI - Tissue-engineered heart valves develop native-like collagen fiber architecture. AB - Creating autologous tissues with on-demand and native-like biomechanical properties is the ultimate challenge in functional heart valve tissue engineering. A promising approach toward this goal is to induce development of native-like tissue structure in vitro by mimicking the diastolic loading phase in a bioreactor. Heart valves cultured with this approach showed in vitro sufficient strength to withstand systemic pressures. This study aims to link global functioning of these valves to the development of a native-like fiber architecture induced by in vitro diastolic loading. It is hypothesized that increased loading magnitude during culture will lead to increased collagen fiber alignment. To test this hypothesis, 10 tissue-engineered heart valves were subjected to different loading protocols in vitro. Local fiber distribution and mechanics were determined in an inverse numerical-experimental approach, combining indentation tests with confocal imaging. Indentation tests on native ovine heart valves were used as a comparison. Although the effect of loading magnitude was small within the tested range, results indicated that the local fiber architecture indeed developed toward native structural properties for all loading protocols. However, apparent fiber mechanics were much stiffer compared with native. This confirms that in vitro mechanical conditioning induces development of a native-like tissue architecture, which underlines its importance for functional heart valve tissue engineering. PMID- 20001250 TI - Tissue engineering of lung: the effect of extracellular matrix on the differentiation of embryonic stem cells to pneumocytes. AB - We have previously differentiated lung epithelium from human and murine embryonic stem cells (mESCs) and are now exploring the potential applications of these cells, including in the engineering of lung tissue constructs. In this study, we hypothesized that the differentiation and maintenance of lung epithelium derived from mESCs can be enhanced by extracellular matrix (ECM) proteins. Our established differentiation protocol was applied to mESCs grown on a range of ECMs: collagen I, laminin 332, fibronectin, Matrigel, and, as an experimental control, gelatin. The ECMs were coated onto tissue culture plastic (TCP) and poly DL-lactic acid (PDLLA), a biodegradable polymer we have previously shown to support the growth of mature pneumocytes. Matrigel or Laminin-332 coating of either TCP or PDLLA film resulted in enhanced surfactant protein C gene expression in differentiating mESCs, a direct indication of the upregulation of lung epithelial differentiation. For each combination, changes in the contact angle and zeta potential of protein-coated TCP and PDLLA film confirmed protein adsorption. We conclude that the choice of the coating protein can greatly affect the differentiation of ESCs, and laminin-332-coated PDLLA provided an ECM degradable scaffold combination that is suitable for engineering of lung tissue constructs. PMID- 20001251 TI - Thermodynamic effects on organismal performance: is hotter better? AB - Despite decades of research on the evolution of thermal physiology, at least one fundamental issue remains unresolved: whether the maximal performance of a genotype depends on its optimal temperature. One school argues that warm-adapted genotypes will outperform cold-adapted genotypes because high temperatures inevitably accelerate chemical reactions. Yet another school holds that biochemical adaptation can compensate for thermodynamic effects on performance. Here, we briefly discuss this theoretical debate and then summarize empirical studies that address whether hotter is better. In general, comparative and experimental studies support the view that hotter is better. Furthermore, recent modeling has shown that thermodynamic constraints impose unique selective pressures on thermal sensitivity. Nevertheless, the thermodynamic effect on maximal performance varies greatly among traits and taxa, suggesting the need to develop a more sophisticated view of thermodynamic constraints. PMID- 20001252 TI - Alignment-free sequence comparison (I): statistics and power. AB - Large-scale comparison of the similarities between two biological sequences is a major issue in computational biology; a fast method, the D(2) statistic, relies on the comparison of the k-tuple content for both sequences. Although it has been known for some years that the D(2) statistic is not suitable for this task, as it tends to be dominated by single-sequence noise, to date no suitable adjustments have been proposed. In this article, we suggest two new variants of the D(2) word count statistic, which we call D(2)(S) and D(2)(*). For D(2)(S), which is a self standardized statistic, we show that the statistic is asymptotically normally distributed, when sequence lengths tend to infinity, and not dominated by the noise in the individual sequences. The second statistic, D(2)(*), outperforms D(2)(S) in terms of power for detecting the relatedness between the two sequences in our examples; but although it is straightforward to simulate from the asymptotic distribution of D(2)(*), we cannot provide a closed form for power calculations. PMID- 20001254 TI - Principles of biomimetic vascular network design applied to a tissue-engineered liver scaffold. AB - Branched vascular networks are a central component of scaffold architecture for solid organ tissue engineering. In this work, seven biomimetic principles were established as the major guiding technical design considerations of a branched vascular network for a tissue-engineered scaffold. These biomimetic design principles were applied to a branched radial architecture to develop a liver specific vascular network. Iterative design changes and computational fluid dynamic analysis were used to optimize the network before mold manufacturing. The vascular network mold was created using a new mold technique that achieves a 1:1 aspect ratio for all channels. In vitro blood flow testing confirmed the physiologic hemodynamics of the network as predicted by computational fluid dynamic analysis. These results indicate that this biomimetic liver vascular network design will provide a foundation for developing complex vascular networks for solid organ tissue engineering that achieve physiologic blood flow. PMID- 20001257 TI - Clinical features of patients with diabetes mellitus presenting with their first episode of uveitis. AB - PURPOSE: Little is known about uveitis in patients with diabetes mellitus (DM). The authors studied diabetic patients with their first episode of uveitis. METHODS: Cross-sectional, case note study documenting the uveitis, underlying cause/syndrome, treatment, type of DM and treatment, and any diabetic retinopathy. RESULTS: There were 34 patients (M/F: 17/17, 48 eyes) with their first uveitis episode (33 had type 2 DM). Mean age of onset of DM 49 years and uveitis 56 years. Uveitis was bilateral in 14 (45%), with most having idiopathic anterior uveitis. Visual acuity 6/18-6/60 in 15 eyes, and worse than 6/60 in 11 eyes. There was 3-4+ flare in 16 eyes, 3-4+ anterior chamber cells in 13 eyes. Diabetic retinopathy was seen in 20 (42%) eyes, and mean blood glucose was 13.64 mmol/L in 11 patients. CONCLUSIONS: Diabetic patients presenting with uveitis, whatever the aetiology, may have severe inflammation, reduced vision, and poor glycaemic control. PMID- 20001255 TI - The future of primary intraocular lymphoma (retinal lymphoma). AB - Basic science and clinical investigations in cancer research have contributed to our understanding of the genetic causes of various neoplasms and discovery of novel therapeutic interventions to fight malignancies such as lymphoma. During this exciting time, we have witnessed the advent of new technologies to further characterize primary intraocular lymphoma (PIOL), or retinal lymphoma, which is selected as the first "Disease of the Year" by Ocular Immunology and Inflammation. Different comprehensive aspects of PIOL, including epidemiology, clinical manifestations, diagnosis, pathophysiology, therapy, and animal models are discussed. The future of PIOL holds an opportunity to really understand the unique cytologic, histopathologic, physiological and immunologic features, as well as the genotypic traits (gene expression, interaction, polymorphism, epigenetics, etc.) and epidemiology. This information will empower us to truly make a difference in patients' managements with this devastating disease. While most of this technology already exists, much work still needs to be done to make translational therapy a reality for PIOL patients in the future. PMID- 20001256 TI - The immune privileged retina mediates an alternative activation of J774A.1 cells. AB - PURPOSE: We have previously found that retinal pigment epithelial (RPE) cells suppressed endotoxin-stimulated macrophages; moreover, it induced expression of anti-inflammatory cytokines. We further assessed the possibility that the RPE is alternatively activating macrophages. METHODS: J774A.1 cells were stimulated with endotoxin and treated with the conditioned media (CM) of RPE, or neuroretinal eyecups from healthy mouse eyes. The supernatant was assayed for IL-1 beta, TNF alpha, IL-6, IL-12(p70), and IL-10, and for nitric-oxide generation. The RPE conditioned media (RPE CM) was absorbed of known soluble factors to identify the factor that augments nitric-oxide generation. RESULTS: We found the RPE CM suppressed all cytokine production except IL-10, and augmented nitric-oxide generation. The augmented nitric-oxide levels were mediated by RPE derived alpha melanocyte stimulating hormone (alpha-MSH). CONCLUSIONS: Healthy RPE not only suppresses inflammatory activity, it promotes an alternative activation of macrophages that can further promote immune privilege. PMID- 20001258 TI - Retinal phlebitis associated with autoimmune hemolytic anemia. AB - PURPOSE: To describe a case of retinal phlebitis associated with autoimmune hemolytic anemia. DESIGN: Observational case report. METHODS: A 44-year-old Indian man diagnosed with autoimmune hemolytic anemia presented with a 1-week history of blurred vision in both eyes. Fundus biomicroscopy revealed bilateral peripheral retinal venous sheathing and cellophane maculopathy. RESULTS: Fundus fluorescent angiogram showed bilateral late leakage from the peripheral venous arcades and submacular fluid accumulation. The retinal phlebitis resolved following a blood transfusion and administration of systemic steroids. CONCLUSIONS: Retinopathy associated with autoimmune hemolytic anemia is not well known. This is thought to be the first documentation of retinal phlebitis occurring in this condition. PMID- 20001259 TI - Culture proven endogenous bacterial endophthalmitis in apparently healthy individuals. AB - PURPOSE: To report 7 eyes of 7 patients who were diagnosed as the cases of endogenous endophthalmitis and proved by culture reports. Though in most of the previously published series, one or more predisposing factors were present in the patients of endogenous endophthalmitis, the authors are presenting a case series of culture-proven endogenous endophthalmitis in apparently healthy and immunocompetent individuals. DESIGN: Retrospective noncomparative case series. All patients were diagnosed as endogenous bacterial endophthalmitis with positive vitreal culture. Variables studied were demographic characteristics, microbiology, therapeutic interventions done, final visual and anatomical outcome. RESULTS: Streptococcus pneumoniae was the most frequent isolate 43% (3 out of 7 eyes) followed by Staphylococcus aureus and E. coli in equal frequency 29% (2 each out of 7 eyes). The median time between the onset of symptoms and presentation with features of endophthalmitis to hospital was 7 days. The presenting visual acuity was less than HM (hand motions) in all cases except one, which had VA of 20/200. No single underlying predisposing condition was identifiable in any of the cases. All the cases were given intravitreal therapeutic agents (one or more doses of antibiotics and steroid) and pars plana vitrectomy was performed in 2 cases. The final visual acuity was not better than presenting VA except in 1 case, which had final VA of 20/40. Intraocular pressure was on the lower side (mean IOP 7 mmHg compared to 12 mmHg in the fellow eye). No eye was enucleated or eviscerated. CONCLUSION: Though in most of the previously published literature endogenous endophthalmitis has been a metastatic ocular infection, the present study describes a series of endogenous bacterial endophthalmitis de novo in onset, without any identifiable predisposing factors. The overall age of presentation was in a younger population than in previously published series. The overall visual outcome was poor, probably due to the serious nature of disease itself and the relatively late presentation. PMID- 20001260 TI - Autofluorescence imaging in rubella retinopathy. AB - PURPOSE: To demonstrate that fundus autofluorescence (FAF) imaging aides the diagnosis of rubella retinopathy (RR). METHODS: Four patients with sensorineural hearing loss underwent ophthalmic exams, electro-retinogram testing (ERG), and FAF in both eyes. Fluorescein angiogram (FA) was performed in one case. RESULTS: Salt-and-pepper retinopathy was observed in seven eyes. FAF highlighted the fundus abnormalities with stippled fluorescence in all seven eyes. ERG showed normal retinal function in all eyes. Choroidal neovascularization was identified by FA in one eye. CONCLUSIONS: FAF is sensitive to the retinal pigment epithelium disturbance caused by rubella and noninvasively helps establish the diagnosis of RR. PMID- 20001261 TI - Clinical review: Anti-TNFalpha therapies in uveitis: perspective on 5 years of clinical experience. AB - Despite a lack of robust evidence, anti-TNF therapies are in wide use for the treatment of noninfectious ocular inflammatory diseases. There is a clear rationale, based on mechanistic and preclinical efficacy data, for their use in posterior segment intraocular inflammation. However, their increasing use for other indications has been largely extrapolated from the benefit observed in autoinflammatory and autoimmune systemic diseases. Given their cost and the potential for significant adverse events, this review highlights the evidence for their continued use, possibilities for switching anti-TNF agents, and ways of reducing the risk of therapy. PMID- 20001262 TI - Mycophenolate sodium for immunosuppressive treatment in uveitis. AB - PURPOSE: To assess the efficacy and tolerability of mycophenolate sodium (MPS) in patients with uveitis. METHODS: Retrospective analysis including uveitis patients treated with MPS (Myfortic) for at least 3 months duration. MPS was administered in a dose of 720 mg twice daily. RESULTS: We analyzed 35 patients (65 affected eyes) with anterior (n = 5), intermediate (n = 23), posterior (n = 6), and panuveitis (n = 1). Previous treatment consisted of systemic corticosteroids in all patients and at least one immunomodulating agent in 15 patients. Mean duration of MPS therapy was calculated as 9.6 months (3-31 months). MPS was able to control intraocular inflammation without relapse in 30 patients (85.7%). Stabilization or improvement of visual acuity was achieved in 60 eyes (92.3%). Tolerability of MPS was good or moderate in 34 patients (97.1%). CONCLUSIONS: MPS was demonstrated as an effective and well-tolerated immunosuppressive drug for different forms of uveitis. PMID- 20001263 TI - Trabeculectomy with mitomycin C in refractory glaucoma associated with nonnecrotizing anterior scleritis. AB - We report a case of refractory secondary-glaucoma associated with scleritis managed successfully with Mitomycin-C augmented trabeculectomies and preoperative transscleral cyclodiode laser(TSCL). Intraocular pressures (IOPs) were uncontrolled on maximal medical therapy (70mmHgOU).Primary emergency TSCL was performed using reduced parameters whilst optimal immunosuppression was achieved. Uncomplicated elective secondary MMC-trabeculectomies (MMC 0.2mg/ml) were later performed. Over four years postoperative follow-up IOPs have remained within normal range without IOP-lowering medication or bleb manipulations (IOP range 7 14mmHgOU).No scleral melt, scleromalacia or scleritic episodes have occurred postoperatively. This case illustrates that MMC-trabeculectomy and TSCL can be used safely and effectively with appropriate immunosuppression in scleritis associated secondary glaucoma [corrected]. PMID- 20001265 TI - Fluocinolone acetonide intravitreal implants in Vogt-Koyanagi-Harada disease. AB - PURPOSE: To describe the use of fluocinolone acetonide implants (Retisert) in Vogt-Koyanagi-Harada disease (VKH). DESIGN: Interventional case series. METHODS: Retrospective review of medical records. RESULTS: Two patients with VKH requiring high-dose systemic corticosteroid therapy to control their inflammation and bilateral serous retinal detachments received bilateral fluocinolone acetonide implants. Upon tapering of systemic corticosteroids, one patient had recurrent serous retinal detachments and the other patient's anterior chamber and vitreous inflammation returned. CONCLUSIONS: The authors' experience with fluocinolone acetonide implants in VKH has been mixed with an inability to fully taper off of systemic corticosteroids. PMID- 20001264 TI - Short-term results of a single intravitreal bevacizumab (avastin) injection versus a single intravitreal triamcinolone acetonide (kenacort) injection for the management of refractory noninfectious uveitic cystoid macular edema. AB - PURPOSE: To report the short-term results of a single intravitreal injection of bevacizumab (IVB) versus a single intravitreal injection of triamcinolone acetonide (IVT) to treat refractory noninfectious uveitic cystoid macular edema (CME). METHODS: Twenty-eight consecutive patients (36 eyes) were retrospectively included. Patients received either 2.5 mg of IVB (16 eyes) or 4 mg of IVT (20 eyes). RESULTS: In the IVT group, baseline best-corrected visual acuity (BCVA) was logMAR 1.1 +/- 0.2, and improved to 0.7 +/- 0.3 (p < .001) at 6 months. In the IVB group, baseline BCVA was logMAR of 1.2 +/- 0.4 and improved to 0.8 +/- 0.4 at 6 months (p = .031). At 6 months, central macular thickness (CMT) in the IVT group improved from 454.8 +/- 238.9 microm to 296 +/- 134.4 microm (p < .0001). CONCLUSION: A single IVT injection improves BCVA and reduces CMT more effectively than IVB in refractory noninfectious uveitic CME at 6 months. PMID- 20001268 TI - Multilineage differentiation potential of fibroblast-like stromal cells derived from human skin. AB - Adult stem cells that reside in adult tissues have been deemed to possess great potential for clinical application because of their capabilities of self-renewal and differentiation. However, the limitations such as infection risks and low isolation rate make the search for appropriate source to be continued. Here, we demonstrate isolation of progenitors from human foreskin tissue samples, which have fibroblast-like morphology and could be easily propagated for more than 50 passages. These foreskin-derived fibroblast-like stromal cells (FDSCs) expressed CD90, CD105, CD166, CD73, SH3, and SH4, which is similar to the immunophenotypes of human bone marrow-derived mesenchymal stem cells. In comparison with Hs68, the human fibroblast cell line, FDSCs are positive for CD105 and absent for CD54 expression. Further, FDSCs could be induced to differentiate into osteocytes, adipocytes, neural cells, smooth muscle cells, Schwann-like cells, and hepatocyte like cells. Interestingly, when cultured in Dulbecco's modified Eagle's medium/F12 medium, FDSCs can form spheres with increased expression levels of fibronectin and vimentin. In conclusion, foreskin can serve as a valuable source of multipotent cells with the capabilities for endodermal, mesodermal, and ectodermal cells. Coupled with the advantages of their easy access, isolation, and propagation, these foreskin-derived stromal cells might be of potential use in future studies in stem cell differentiation and therapeutic application. PMID- 20001269 TI - Controlling medium osmolality improves the expansion of human articular chondrocytes in serum-free media. AB - To investigate the effects of medium osmolality on the expansion of human articular chondrocytes (HACs) with serum-free media (SFM), proprietary SFM of various osmolalities (290, 320, 350, 400, and 450 mOsm/kg), supplemented with components known to enhance chondrocyte growth, were constructed by the adjustment of NaCl concentration. It was found that HACs obtained better expansion in SFM at osmolalites lower than the average osmolality (400 mOsm/kg) of human articular cartilage in vivo. SFM at 290, 320, and 350 mOsm/kg showed similar growth, attaining up to a 1.55-fold increase in the proliferation rate compared with SFM at 400 mOsm/kg. Increasing SFM osmolality to 450 mOsm/kg resulted in a proliferation rate of 0.65-fold lower than at 400 mOsm/kg. Chondrogenic capacity was also examined via three-dimensional pellet cultures in a chondrogenic medium with HACs expanded in 320 and 400 mOsm/kg SFM. Biochemical, histological, and immunohistochemical analyses revealed similar glycosaminoglycan and collagen type II contents in both groups. Taken together, these results show that the expansion of HACs in SF cultures can be improved by adjusting the medium osmolality to be within the range of 290-350 mOsm/kg and that controlling medium osmolality during monolayer cultures does not deter the tissue-forming capability of the cells. PMID- 20001270 TI - Alginate encapsulation as a novel strategy for the cryopreservation of neurospheres. AB - Primary cultures of brain cell neurospheres are valuable in vitro models for neurotoxicology and brain cell research. Such applications would greatly benefit from the development of efficient cryopreservation protocols that assure the availability of viable and genetically stable stocks of functional neurospheres. In this work we aimed at developing an integrated strategy allowing for long-term culture and cryopreservation of brain cell neurospheres with high viability and reduced recovery time postthawing. Microencapsulation in clinical-grade, ultrahigh viscous, highly purified alginate uniformly cross-linked with Ba(2+) was evaluated as the main strategy to avoid the commonly observed loss of cell cell and cell-matrix interactions with consequent aggregate's fragmentation and decrease in cell viability that occurs postthawing. Brain cells isolated from 16 day-old fetal rats were cultured in spinner vessels as neurospheres, encapsulated at the 5th day of culture, and cryopreserved at day 19. Culture characterization and assessment of postthawing recovery, concerning cell metabolism, aggregate's cell type composition, and neuron-astrocyte interactions were performed through analysis of membrane integrity, metabolic activity assays, and immunohistochemistry. Our results show that the encapsulation process does not affect cell viability's central metabolism; neither cell differentiation nor cell extensions into cell networks are usually observed between neurons and astrocytes within the neurosphere structure. Neurosphere encapsulation resulted in reduced recovery time postthawing and significantly less fragmentation. Further, the use of serum-free CryoStorTM solution provided further protection for both nonencapsulated and encapsulated aggregates compared with serum-supplemented culture medium as the cryopreservation medium. PMID- 20001271 TI - Preparation, in vitro characterization and in vivo release of naproxen loaded in poly-caprolactone nanoparticles. AB - Naproxen was loaded in poly-caprolactone (PCL) nanoparticles as an implantable sustained release system to prolong its anti-inflammatory activity. Naproxen loaded nanoparticles were produced with the following characteristics: Nanometric size (< 300 nm), negative zeta potential, low polydispersity index (< 0.1), satisfactory encapsulation efficiency, low water content (< 1%), and spherical shape. In vitro naproxen release profile was sustained and the kinetic followed the Higuchi model. The PCL nanoparticles containing about 12.5% (w/w) of the naproxen (sample A3) was chosen for complementary studies of stability and in vivo release in rats. Nanoparticles did not suffer alteration during stability studies. In vivo release was sustained by one month. Thus, nanoparticles showed potential to act as an implantable sustained release system for chronic inflammatory diseases use. PMID- 20001272 TI - Combination of adoptive cell transfer and antibody injection can eradicate established tumors in mice--an in vivo study using anti-OX40mAb, anti-CD25mAb and anti-CTLA4mAb-. AB - Boosting an effective immune response against established tumors remains a difficult challenge. This study shows the combination of 1) adoptive cell transfer using CD25 depleted splenocytes co-cultured with irradiated tumor cells, and 2) antibody injection therapy using CTLA4 blockade, the elimination of Treg and OX40, which together could eradicate an established MethA tumor in over 50% of the BALB/c mice. Each element of the protocol was shown to be necessary, as elimination of any factor except anti-CD25 antibody injection failed to eradicate the tumor. PMID- 20001273 TI - Immunomodulatory properties of exogenous surfactant in adult rat alveolar macrophages. AB - BACKGROUND AND AIM: The study investigates the immune response of the lung alveolar macrophages in adult rats with intratracheal instillation of surfactant, isolated from porcine, rabbit and rat lung as well as the synthetic surfactant. MATERIAL AND METHODS: Adult male rats were divided into five groups: Controls were instilled with buffer, similarly P-SF, R-SF, r-SF, and S-SF group of animals were instilled with porcine, rabbit, rat and synthetic surfactant, respectively. RESULTS: After one month, IL-1beta secretions were found to be elevated in the supernatant of alveolar macrophages. Further, nitric oxide (NO) level was elevated with porcine and synthetic surfactant, while reactive oxygen species (ROS) generation was found to be inhibited significantly in all the treatment groups. Apoptoses were studied by using fluorescence dyes acridine orange/ethidium bromide, where percent of the apoptotic cells were found to be decreased in all the treated groups. CONCLUSIONS: Exogenous surfactant elevates secretion of pro-inflammatory cytokine IL-1beta and also the NO as signaling molecule. The inhibitory effects of surfactants on ROS generation and on apoptosis may result from the interruption in the cell signaling pathway. In the treatment of respiratory distress syndrome, the anti-inflammatory response of surfactant may thus depend upon the specific preparation of the surfactant. PMID- 20001274 TI - Synthesis and biological activity of pyrazolo[3,4-d]thiazolo[3,2-a]pyrimidin-4 one derivatives: in silico approach. AB - Xanthine oxidase (XO) is responsible for the pathological condition called gout. Inhibition of XO activity by various pyrazolo[3,4-d]thiazolo[3,2-a]pyrimidine-4 one derivatives was assessed and compared with the standard inhibitor allopurinol. Out of 10 synthesized compounds, two compounds, viz. 3-amino-6-(2 hydroxyphenyl)-1H-pyrazolo[3,4-d]thiazolo[3,2-a]pyrimidin-4-one (3b) and 3-amino 6-(4-chloro-2-hydroxy-5-methylphenyl)-1H-pyrazolo[3,4-d]thiazolo[3,2-a]pyrimidin 4-one (3g) were found to have promising XO inhibitory activity of the same order as allopurinol. Both compounds and allopurinol inhibited competitively with comparable Ki (3b: 3.56 microg, 3g: 2.337 microg, allopurinol: 1.816 microg) and IC(50) (3b: 4.228 microg, 3g: 3.1 microg, allopurinol: 2.9 microg) values. The enzyme-ligand interaction was studied by molecular docking using Autodock in BioMed Cache V. 6.1 software. The results revealed a significant dock score for 3b (-84.976 kcal/mol) and 3g (-90.921 kcal/mol) compared with allopurinol (-55.01 kcal/mol). The physiochemical properties and toxicity of the compounds were determined in silico using online computational tools. Overall, in vitro and in silico study revealed 3-amino-6-(4-chloro-2-hydroxy-5-methylphenyl)-1H pyrazolo[3,4-d]thiazolo[3,2-a]pyrimidin-4-one (3g) as a potential lead compound for the design and development of XO inhibitors. PMID- 20001275 TI - Intranasal drug delivery of didanosine-loaded chitosan nanoparticles for brain targeting; an attractive route against infections caused by AIDS viruses. AB - The primary aim of this study was to investigate intranasal (i.n.) administration as a potential route to enhance systemic and brain delivery of didanosine (ddI). A further aim was to investigate the potential use of chitosan nanoparticles as a delivery system to enhance the systemic and brain targeting efficiency of ddI following i.n. administration. Didanosine-loaded chitosan nanoparticles, were prepared through ionotropic gelation of chitosan with tripolyphosphonate anions, and characterized in terms of their size, drug loading, and in vitro release. The nanoparticles were administered i.n. to rats, compared to i.n. and intravenous (i.v.) administration of ddI in solution. The concentrations of ddI in blood, CSF, and brain tissues were analyzed by ultra performance liquid chromatography mass spectroscopy (UPLC/MS). The brain/plasma, olfactory bulb/plasma and CSF/plasma concentration ratios were significantly higher (P < 0.05) after i.n. administration of ddI nanoparticles or solution than those after i.v. administration of didanosine aqueous solution. The ratio of ddI concentration values of the nanoparticles to the solution at 180 min post-i.n. dosing was 2.1 and 1.9 in CSF and brain, respectively. Thus, both the i.n. route of administration and formulation of ddI in chitosan nanoparticles increased delivery of ddI to CSF and brain. PMID- 20001277 TI - Treatment outcome in children with tuberculosis in southern Ethiopia. AB - Our study took place at 8 tuberculosis diagnostic and treatment centres in Sidama Zone, southern Ethiopia. Our objectives were to assess the status of TB treatment in children and risk factors associated with unfavourable outcome. A retrospective study was based on information retained in TB registers of the centres. Demographic characteristics and treatment outcome of children registered between 2002 and 2007 were retrieved. Risk factors for unfavourable outcome (failure, default or death) were calculated. Of 851 (165 with smear-positive, 475 smear-negative and 206 extrapulmonary TB) children, 655 (77%) were cured or completed and 124 (14.6%) had unfavourable outcome. Treatment success rate increased with age from 66% in children <5 y old to 81% in 5-9-y-olds and 85% in >/=10-y-olds (p <0.001). 75% of patients with smear-negative TB had favourable outcome compared to 80.6% for smear-positive cases (p =0.06). Age <5 y, lack of smear conversion in the second month and living in rural areas were independent risk factors for unfavourable treatment outcome. In conclusion, the outcome of TB treatment varies with age, residency and smear results. Reporting only cases with smear-positive TB disproportionally excludes a higher proportion of children than adults. Surveillance systems to monitor TB outcome in children need to be improved. PMID- 20001276 TI - Treatment of hepatitis C virus infection: updated Swedish Consensus recommendations. AB - In a recent expert meeting, Swedish recommendations for the treatment of HCV infection were upgraded. The panel recommends vaccination against both hepatitis A and B in patients with HCV. Therapy for symptomatic acute HCV infection should be initiated if spontaneous resolution has not occurred within 12 weeks, whereas asymptomatic acute HCV should be treated upon detection. Patients with genotype 2/3 infection should generally be treated for 24 weeks. In patients with a very rapid viral response (vRVR), i.e. HCV RNA below 1000 IU/ml on d 7, treatment can be shortened to 12-16 weeks, provided that no dose reduction has been made. For genotype 1 patients with rapid viral response (RVR), 24 weeks treatment is recommended. For patients with a complete early viral response (cEVR), 48 weeks treatment is recommended, whereas 72 weeks treatment should be considered for patients with partial early viral response (pEVR). For patients with difficult-to treat disease and with pronounced anaemia, erythropoietin can be used to maintain the ribavirin dose. In HCV-HIV coinfected patients, combination therapy for HCV should, if possible, be initiated before anti-retroviral therapy (ART) is indicated. For liver transplant patients pre-emptive therapy is not recommended; hence, treatment should be deferred until histological recurrence. PMID- 20001279 TI - Line probe assay for differentiation within Mycobacterium tuberculosis complex. Evaluation on clinical specimens and isolates including Mycobacterium pinnipedii. AB - A line probe assay (GenoType MTBC) was evaluated for species differentiation within the Mycobacterium tuberculosis complex (MTBC). We included 387 MTBC isolates, 43 IS6110 low-copy MTBC isolates, 28 clinical specimens with varying microscopy grade, and 30 isolates of non-tuberculous mycobacteria. The assay was 100% specific and identified all 387 isolates and 98% of all IS6110 low-copy strains in concordance with the gold standard. The 2% discrepancy was caused by 1 isolate showing a faint restriction fragment length polymorphism (RFLP) pattern. The assay could provide specifies identification in 13 of 19 (68%) microscopy positive specimens and 0 of 9 microscopy-negative specimens. To our surprise, the probe for M. africanum subtype I reacted with M. pinnipedii. This cross-reaction has not previously been reported. The assay was rapid, easy to perform and directly applicable in highly smear-positive specimens. We predict that the assay will enable enhanced surveillance of species-specific treatment outcome, which may change treatment regimens. PMID- 20001281 TI - Seroprevalence of HIV, HSV-2, and Treponema pallidum in the Kosovarian population. AB - The objective of this study was to evaluate the seroprevalence of infection with HIV, herpes simplex virus type 2 (HSV-2), and Treponema pallidum (TP) in a Kosovarian population. A cross-sectional study was performed in Peja, Kosovo, from January to March 2005, among 1285 persons recruited at the Peja Hospital. The seroprevalence of HIV, HSV-2, and TP was evaluated, and the viral correlates for each infection were analysed. No HIV-positive cases were found. The seroprevalence of HSV-2 was 20.2%. The factors significantly associated with HSV 2 infection at the multivariate analysis were: female gender (adjusted OR, 1.73; 95% CI 1.24-2.41) and being married (adjusted OR, 1.46; 95% CI 1.06-2.01). Three persons (0.2%) had a positive serology for TP. The only risk factor associated with TP infection was age = 50 y. Our results show a low seroprevalence of HIV infection and TP, and a high seroprevalence of HSV-2 in Kosovo. These findings suggest the need for appropriate surveillance systems, prevention programmes, and information aimed at controlling the spread of HIV and other sexually transmitted infections in this area. Moreover, the circulation of infections acquired through sexual contact may facilitate an increase in the sexually transmitted HIV epidemic in the near future. PMID- 20001282 TI - Drinking alcohol surrogates among clients of an alcohol-misuser treatment clinic in Novosibirsk, Russia. AB - A pilot study was conducted in the Russian city Novosibirsk during 2006-2007 with 40 participants sampled from clients admitted to health services for drinking related problems to explore surrogates for alcoholic beverage drinking using semistructured interviews. Types of surrogates consumed and reasons for their consumption were investigated. Results revealed that the most prevalent reported surrogate used was industrial spirit. The main reasons for drinking surrogates were the high affordability and physical availability of surrogates combined with the need to relieve severe withdrawal symptoms. The study limitations are described, and suggestions for future research are made. PMID- 20001283 TI - Temperamental risk factors for adolescent cannabis use: a systematic review of prospective general population studies. AB - In order to establish the evidence for prospective relationships between temperamental and personality indicators of behavioral undercontrol and adolescent cannabis use, we systematically searched relevant papers published through April 2008. We assessed and evaluated 14 studies, of which only 4 were considered of high quality. Using "best evidence" synthesis, we found weak to moderate evidence for prospective relations between a combination of high approach and low avoidance and several measures of cannabis use. The study's limitations are noted. This review provides suggestions and recommendations for future studies in this area. PMID- 20001284 TI - Recruiting hard-to-reach drug-using men who have sex with men into an intervention study: lessons learned and implications for applied research. AB - Drug (ab)use researchers and service providers across the globe have been challenged with locating target populations and subsequently enrolling participants into their programs. This study presents data from nearly 3 years (2004-2006) of recruiting "high-risk" drug-using gay and bisexual men into a clinical research trial based in New York City. During the enrollment period, two recruitment/marketing strategies were utilized: (1) marketing of the intervention research study itself to men who were in the early stages of identifying problems with their drug use and risky sexual behavior and (2) two-stage recruitment via a lower-threshold/commitment (i.e., brief survey) and subsequent offering/enrollment into the full trial upon completion of the initial visit (i.e., a foot-in-the-door). The second approach was substantially more effective in enrolling participants into the full trial (6.3 participants/month vs. 2.5 participants/month). Furthermore, recruitment costs for the foot-in-the-door approach were substantially reduced ($356.57 per participant vs. $497.03 per participant). Compared to the marketing of interventions themselves to target populations, a two-stage recruitment strategy incorporating lower-threshold interactions may be a more effective approach to recruit for interventions. PMID- 20001285 TI - Transitions to substance use and sexual intercourse among South african high school students. AB - This study examined longitudinal patterns of initiating substance use and sexual intercourse among a sample of 1,143 high school students from a low-income township in Cape Town, South Africa. Longitudinal data on lifetime incidence of sexual intercourse and alcohol and marijuana use were collected semiannually from 2004 to 2006. Latent transition analysis (LTA) was used to test competing models of transitions to these behaviors. Participants were more likely to move from no risk behavior to substance use, rather than from no risk behavior to sexual intercourse. At all time points, the proportion of youth who had engaged in sexual intercourse but not substance use was very small. The study's limitations and directions for future research are discussed. PMID- 20001286 TI - Tolerance and sensitization to the effects of cocaine use in humans: a retrospective study of long-term cocaine users in Philadelphia. AB - In the effort to develop medications to combat addiction, researchers have developed models that attempt to describe the neurobiological process of cocaine dependence. It has not, however, yet been determined which of these models, if any, best fits the behaviors and experiences of patients. This project retrospectively evaluated changes in patients' experiences with cocaine over time in order to clarify the model that best fits clinical observations. In 2005 and 2007, 100 treatment-seeking, long-term cocaine users were recruited from an urban university-based treatment center in Philadelphia, PA, United States. Each participant was administered the "Cocaine History Questionnaire" which asked them to describe the initiation and escalation of their cocaine usage, changing reward perceptions, and effects of intoxication at certain points in their drug use careers. This data was then analyzed using repeated measures, examining the within subject differences in reported information over the time points. We found evidence that while the amount of drug used increases, self-reported euphoria decreases while negative symptoms associated with cocaine use also increase. The data provide preliminary evidence for the hedonic dysregulation model of addiction. Limitations and implications of the study are discussed in the conclusion. PMID- 20001287 TI - The influence of age and gender on party drug use among young adults attending dance events, clubs, and rock festivals in Belgium. AB - A random sample of visitors of dance events, clubs, and rock festivals in Belgium (Flanders) was selected to complete an anonymous survey regarding their use of "party" drugs (alcohol, cannabis, xtc, cocaine, and amphetamines) and patterns of going out. The results of 670 respondents recruited in 2005 are reported and compared with 2003 for gender and age. Drug use in these nightlife settings is higher than in the general population. In younger age groups, the illegal drug use increases, but it decreases in older age groups. This might be linked to more responsibilities. Men still use drugs more often than women, but the gender differences seem to decrease. The changing role of women in society could be an explanation for this evolution. Limitations and implications of this research are discussed. PMID- 20001288 TI - Follow-up of provision of inpatient treatment for UK Healthcare Professionals with alcohol dependence: snapshot of a pilot specialist National Health Service. AB - Alcohol dependent healthcare professionals (AHCPs) entering a specialized National Health Service (NHS) inpatient treatment facility in southeast London, UK, were investigated. A retrospective case-note analysis was conducted, supplemented with a postal questionnaire. Twenty-three mature (mean age 46.7 years; 13 male) AHCPs, 82% of admissions, with a mean duration of alcohol dependence of 16.5 years were followed-up at an average of 26 months postdischarge. Half of the sample met criteria for lifetime psychiatric disorder. Physical illness was also a prominent problem. At follow-up, six professionals reported continuous abstinence since discharge. For those who continued drinking, there was a significant reduction in daily quantity consumed. This study suggests that the absence of an adequate treatment infrastructure and administrative resources have a major impact on access to treatment services and outcomes for addicted healthcare professionals. There is a need for supported and dedicated services for this group within the UK National Health Service. The study's limitations are noted. PMID- 20001289 TI - Do spirituality and religiosity help in the management of cravings in substance abuse treatment? AB - The purpose of this study was to examine the relationship of spirituality, religiosity and self-efficacy with drug and/or alcohol cravings. A cross sectional survey was completed by 77 male participants at an Australian Salvation Army residential rehabilitation service in 2007. The survey included questions relating to the participants' drug and/or alcohol use and also measures for spirituality, religiosity, cravings, and self-efficacy. The sample included participants aged between 19 and 74 years, with more than 57% reporting a diagnosis for a mental disorder and 78% reporting polysubstance misuse with alcohol most frequently endorsed as the primary drug of concern (71%). Seventy five percent of the clients reported that spirituality and religious faith were useful components of the treatment program. A multivariate multiple regression analysis identified that spirituality and self-efficacy have significant relationships with cravings. Self-efficacy mediated the relationship between spirituality and drug and/or alcohol cravings. The limitations of this study included its cross-sectional design and a sample that was drawn from a faith based program. Future research would benefit from the longitudinal examination of the relationship between spirituality, self-efficacy, and cravings; the exploration of a broader range of client-specific and interpersonal variables; and the inclusion of a control group from a secular treatment facility. PMID- 20001291 TI - Incidence and predictors of onset of injection drug use in a San Francisco cohort of homeless youth. AB - Few studies document incidence of injection drug use among homeless youth. We followed a cohort of 70 street-recruited homeless youth in San Francisco, California who had never injected drugs for six months in 2004-5. We examined initiation of injection drug use and its predictors, informed by prior ethnographic findings. Data were analyzed using exact logistic regression. 11.4% of youth initiated injection drug use. Having no high school education, being over 21 years old, and being in disequilibrium predicted initiation. Limitations, implications and suggestions for future research are noted. Funding was provided by the National Institute for Child Health and Development. PMID- 20001292 TI - A dialogue on the incapability/capability of injection drug users. PMID- 20001290 TI - Neuropsychological assessment of current and past crack cocaine users. AB - BACKGROUND: Cognitive changes due to crack cocaine consumption remain unclear. METHODS: For clarification, 55 subjects were assigned to three groups: control group, crack cocaine current users, and ex-users. Participants were submitted to Mini-Mental State Examination (MMSE) and tasks evaluating executive functioning and verbal memory. Mood state was also measured. Intergroup comparisons were carried out. RESULTS: Control group performance on the MMSE was better than that of users and ex-users. Verbal memory performance for logical memory of users was impaired. Ex-users scored lower on DSST and Trail Making Test (Part B). CONCLUSION: Chronic crack cocaine use seems to disrupt general cognitive functioning (MMSE), verbal memory, and attentional resources, but findings suggest that some of these effects could be reversed by abstinence. PMID- 20001293 TI - Breast cancer screening: science, society and common sense. PMID- 20001294 TI - Antiproliferative activity of Larrea divaricata Cav. on lymphoma cell line: participation of hydrogen peroxide in its action. AB - Larrea divaricata is a plant used in Argentina. The aim was to analyze the participation of H2O2 in the antiproliferative effect of L divaricata on BW5147 cells. Different studies were performed: proliferation, viability, nitrite production, apoptosis, oxygen reducing activity, NF-kappa B translocation, H2O2 production, H2O2 scavenging activity, and effect on exogenous superoxide dismutase. The extract decreased proliferation and induced apoptosis through P-38 and ERK pathways. The extract activated Mn2+-SOD, increasing H2O2, which was implicated in the antiproliferative mechanism by the increase of Nitric oxide (NO). New drugs, which can increase H2O2, could be a therapeutic strategy for the treatment of cancer. PMID- 20001295 TI - MMP-9 (gelatinase B) expression is associated with disease-free survival and disease-specific survival in colorectal cancer patients. AB - Extracellular matrix degradation is required for invasion and metastasis formation in colorectal carcinoma (CRC), therefore, we have examined matrix metalloproteinases MMP-9 expression in tumors from patients with CRC. The study comprises of 360 patients who underwent bowel resection for stage II, III, IV tumors. Paraffin-embedded CRC tissue samples were used for immunohistochemical staining. Negative MMP-9 expression levels correlated with longer survival time as evaluated by disease-free survival and disease-specific survival (p =.023, p =.006). In multivariate survival (Cox) analysis, MMP9 was a significant independent predictor of DFS (p =.014), but not of DSS, which was independently predicted by disease recurrence, stage and localization. The detection of MMP-9 expression may be valuable in finding patients who are at high risk of developing disease recurrence. PMID- 20001296 TI - Cancer patients and awareness of venous thromboembolism. AB - Cancer-associated thrombosis leads to morbidity and mortality in cancer patients. We sought to ascertain awareness of venous thromboembolism (VTE) and receptiveness to thromboprophylaxis amongst ambulatory cancer patients. In all 190 patients completed the survey over a three-week period. Of these, 100 patients (53%) reported unawareness of the increased risk of thrombosis with malignancy. Of the examined patients, 161 (86%) were willing to use oral anticoagulant for prophylaxis, and 86 (46%) were willing to perform daily anticoagulant injections. This study reveals an alarming lack of knowledge amongst patients regarding the increased risk of thrombosis. Informed patients are willing to use prophylaxis, if its effectiveness is established. PMID- 20001297 TI - Low-dose metronomic paclitaxel chemotherapy suppresses breast tumors and metastases in mice. AB - This study investigated the use of low-dose metronomic (LDM) chemotherapy with paclitaxel in a highly metastatic mouse model of 4T1 breast cancers, and compared it with the maximum tolerable dose (MTD) therapy. LDM therapy displayed a stronger anti-tumor activity in suppressing primary and metastatic breast tumors with less degree of side effects, and stronger anti-angiogenic and anti lymphangiogenic activities than MTD therapy. But MTD therapy showed stronger pro apoptotic and anti-proliferative activities in situ. Paclitaxel therapy downregulated expression of vascular endothelial growth factor receptor-2 (VEGFR2) and up-regulated expression of thrombospondin-1. The results support the application of paclitaxel LDM therapy to treat advanced breast cancer. PMID- 20001298 TI - Bevacizumab may be active in myelofibrosis. AB - A case of a 69-year-old woman with myelofibrosis presenting with non-small cell lung cancer was reported at Sotiria General Hospital in Athens. She was administered bevacizumab infusions biweekly and a great decrease in the number of her white blood cells was observed. Such low levels of white blood cells had never been reached in the past, when she was receiving hydroxyurea. PMID- 20001299 TI - Editorial introduction. The 12 years of evolutionary computation in Dortmund. PMID- 20001300 TI - Editorial Introduction to the Special Issue: Evolutionary computing in the collaborative research centre on computational intelligence at Technische Universitat (TU) Dortmund. PMID- 20001301 TI - Tracheal agenesis with bifurcating common airway arising from midesophagus. PMID- 20001302 TI - Selected papers from the meetings of the International Clinical Phonetics and Linguistics Association. Editorial. PMID- 20001303 TI - Grammatical constructions in Cri du chat syndrome--findings from a case study. AB - The literature on grammatical skills in persons with Cri du chat syndrome (CCS) is very limited, and the need for more knowledge in this area is thus evident, in particular for speech and language therapists working with individuals with this syndrome. This case study report describes the syntactic skills of a 14-year-old Norwegian girl with CCS. The theoretical framework is construction grammar. Data for the study were collected in a diary by the author over a period of 4 months and make up a corpus of 552 utterances. These utterances are described in terms of MLU, diversity of argument structure constructions, proportion and types of complex utterances, use of auxiliaries, as well as deviant word order patterns, types of omissions, and use of prefabricated units. The primary aim of the study is to identify strengths and weaknesses in syntactic skills, which provide the basis for future research on grammatical skills in persons with CCS. PMID- 20001304 TI - Semi-spontaneous oral text production: measurements in clinical practice. AB - Functionally relevant assessment of the language production of speakers with aphasia should include assessment of connected speech production. Despite the ecological validity of everyday conversations, more controlled and monological types of texts may be easier to obtain and analyse in clinical practice. This article discusses some simple measurements for the analysis of semi-spontaneous oral text production by speakers with aphasia. Specifically, the measurements are related to the production of verbs and nouns, and the realization of different sentence types. The proposed measurements should be clinically relevant, easily applicable, and linguistically meaningful. The measurements have been applied to oral descriptions of the 'Cookie Theft' picture by eight monolingual Norwegian speakers, four with an anomic type of aphasia and four without any type of language impairment. Despite individual differences in both the clinical and the non-clinical group, most of the measurements seem to distinguish between speakers with and without aphasia. PMID- 20001305 TI - Repair sequences in dysarthric conversational speech: a study in interactional phonetics. AB - This paper presents some findings from a case study of repair sequences in conversations between a dysarthric speaker, Chris, and her interactional partners. It adopts the methodology of interactional phonetics, where turn design, sequence organization, and variation in phonetic parameters are analysed in unison. The analysis focused on the use of segmental and prosodic variation found during attempts by Chris to repair a previously identified trouble source. The results indicate that trouble sources were extremely common in the recorded conversations, but that repair attempts were almost always communicatively successful. Analysis of the fragments revealed that repair sequences are often collaborative achievements, with the participant's conversational partners signalling the specific trouble source within a turn, or providing feedback about repair attempts. It was also observed that successful repair attempts were not always simple repetitions of the trouble source, but varied in other linguistic areas. It is suggested that intelligibility repairs should be studied using both experimental and qualitative methods. PMID- 20001306 TI - Adult speakers' tongue-palate contact patterns for bilabial stops within complex clusters. AB - Previous studies using Electropalatography (EPG) have shown that individuals with speech disorders sometimes produce articulation errors that affect bilabial targets, but currently there is limited normative data available. In this study, EPG and acoustic data were recorded during complex word final /sps/ clusters spoken by 20 normal adults. A total contact (TC) index measured amount of tongue palate contact during clusters in words such as 'crisps'. Bilabial closure was inferred from the acoustic signal. The TC profiles indicated that normal adults hold their tongues in a steady /s/-like position throughout the cluster; most speakers (85%, n=17) had no significant difference in TC values during bilabial closure compared to flanking fricatives. The results are interpreted as showing that normal speakers produce double bilabial-alveolar articulations for /p/ in these clusters. Although steady state TC profiles were typical of the group, absolute TC values varied considerably between speakers, with some speakers having up to three times more contact than others. These findings add to the knowledge about normal articulation, and will help to improve diagnosis and treatment of individuals with speech disorders. PMID- 20001309 TI - Phonological awareness and phonological hierarchy in unintelligible speech: what does the child really 'know'? AB - Phonological awareness has been claimed to play an important role in the development of language skills, and it is essential that the child's phonological skills be assessed accurately in order to predict and to take early measures to help those that may encounter problems. Although the mora has attracted much attention in the discussion of phonological knowledge and phonological skills of Japanese, to our knowledge there are no previous studies that have assessed disordered speech of Japanese purely from the viewpoint of prosodic levels higher than the mora. In order to fill in this gap, this paper will present data from a child with 'unintelligible speech'. The longitudinal data (4; 11 to 8;6) collected from spontaneous Japanese speech were analysed from the perspective of phonological markedness. The evidence so far indicates that even within severely distorted speech, it may be possible to tease out systematic errors that seem to suggest the existence of unmarked phonological structures such as the bimoraic foot. This finding suggests that in order to accurately assess a child's phonological ability, many different levels within the phonological hierarchy should be taken into consideration. PMID- 20001307 TI - A perceptual and electropalatographic study of /integral/ in young people with Down's syndrome. AB - Speech production in young people with Down's syndrome has been found to be variable and inconsistent. Errors tend to be more in the production of sounds that typically develop later, for example, fricatives and affricates, rather than stops and nasals. It has been suggested that inconsistency in production is a result of a motor speech deficit. Late acquired fricatives such as /s/ and /integral/ are complex articulations, which may require more precise motor programming and may therefore show highly inconsistent productions. Other factors potentially affecting speech production in this population are abnormal palatal structure, hearing loss, and hypotonia. A group of 20 young people with Down's syndrome were recorded using Electropalatography (EPG), reading a wordlist containing the phrase 'a sheep'. The wordlist contained seven other phrases and was repeated 10 times. Eight typically developing, cognitively matched children and eight adults were also recorded producing the same data set. Articulatory (EPG pattern analysis) and perceptual analyses of the 10 productions of /integral/ were carried out. /integral/ production was found to be inconsistent in the young people with Down's syndrome, with more errors both in the auditory analysis and articulatory analysis than in the typical sample, which may be due to a motor programming or motor control problem. There were a greater number of errors in the EPG analysis than in the perceptual analysis. This suggests that some young people with DS were able to produce perceptually acceptable /integral/ with atypical EPG patterns. The use of typical, adult-modelled /integral/ EPG patterns in therapy may be inappropriate for some children with DS who present with atypical palatal structures. PMID- 20001308 TI - Electropalatographic therapy for children and young people with Down's syndrome. AB - Articulation disorders in Down's syndrome (DS) are prevalent and often intractable. Individuals with DS generally prefer visual to auditory methods of learning and may therefore find it beneficial to be given a visual model during speech intervention, such as that provided by electropalatography (EPG). In this study, participants with Down's syndrome, aged 10:1 to 18:9, received 24 individualized therapy sessions using EPG. Simultaneous acoustic and EPG recordings were made pre- and post-intervention during 10 repetitions of a word list containing lingua-palatal consonants. Participants also completed the DEAP phonology sub-test at both time points. Post-treatment, all participants showed qualitative and quantifiable differences in EPG patterns and improvements in DEAP percentage consonants correct. EPG assessment and therapy appears a positive approach for identifying and improving articulatory patterns in children with DS. PMID- 20001310 TI - An interview with Graham White, Ph.D. By Vicki Glaser. PMID- 20001311 TI - [(13)C]Methionine breath test as a marker for hepatic mitochondrial function in HIV-infected patients. AB - Metabolic disturbances such as dyslipidemia, lipodystrophy syndrome, visceral obesity, hyperlactatemia, diabetes mellitus, and hepatic steatosis have been recognized as serious complications in long-term antiretroviral-treated HIV infected patients. The oxidative capacity of liver mitochondria plays a central role in their pathogenesis and can be analyzed using the [(13)C]methionine breath test. We analyzed hepatic mitochondrial function using the [(13)C]methionine breath test in antiretrovirally treated and untreated HIV-infected patients as well as HIV-negative controls. Patients with hepatic steatosis, hypertriglyceridemia, lipohypertrophy, and older age showed reduced methionine metabolism. Hepatic mitochondrial function is impaired in antiretroviral-treated HIV-infected patients with disturbances of lipid metabolism. PMID- 20001312 TI - Dual HIV Infection Uncommon in Patients on Antiretroviral Therapy in a Region with High HIV Prevalence. AB - Dual HIV infection has been found in regions with high HIV prevalence and in populations infected with diverse strains of HIV. The prevalence of dual infection in KwaZulu-Natal (KZN), where there is a high prevalence of HIV and an influx of immigrants who may be infected with different HIV subtypes, is unknown. The aim of this study was to determine the prevalence of dual infection in KZN. Eighty-three samples were obtained from chronically infected patients on ARV treatment from sites throughout KZN. Subtyping of the samples was performed using the heteroduplex mobility assay (HMA). Twelve samples that had possible dual infection by HMA were cloned and sequenced. Phylogenetic analysis showed that there was no dual infection in these samples. Contrary to a previous study, we did not find dual infection in this region. The patients in our study may be different from those in the previous study in terms of transmission risk factors, treatment, and stage of infection (acute vs. chronic). This study may have important implications for vaccine development and for the pathogenesis and treatment of dual HIV infection. PMID- 20001313 TI - Short communication: methicillin-resistant Staphylococcus aureus infections in children and young adults infected with HIV. AB - The epidemiology of methicillin-resistant Staphylococcus aureus (MRSA) infections, in particular with Panton-Valentine leukocidin (PVL)-positive strains, has not been well characterized in children and young adults with HIV infection. It is not known if PVL-positive strains of MRSA cause an increased morbidity in this population compared to PVL-negative strains. The purpose of this study was to retrospectively analyze the epidemiology of PVL-positive and PVL-negative MRSA infections in children and young adults with HIV from 2000 to 2007. Molecular typing was performed by polymerase chain reaction (PCR) for detection of the PVL genes. Staphylococcus Cassette Chromosome (SCC) mec and spa typing were performed on all PVL-positive isolates. The number of HIV patients with MRSA infection increased significantly between 2000 and 2007 ( p=0.0015). Twenty seven (87%) of the 31 MRSA isolates were from skin and soft tissue infections (SSTI). Clindamycin resistance was observed in 19% of the MRSA isolates. PVL-positive isolates bearing the type IV SCC mec element comprised 16 of 31 (52%) MRSA isolates. All the PVL-positive isolates belonged to the USA300 pulsed-field type. There was no difference in the mean CD4 count and HIV viral load between patients with PVL-positive and PVL-negative MRSA infections. PVL positive MRSA infections were associated with more SSTI ( p=0.043) but not with increased morbidity or a higher risk of complications compared to PVL-negative MRSA infections in children and young adults with HIV. PMID- 20001315 TI - Phylogenetic analysis of HIV sequences obtained in a respondent-driven sampling study of men who have sex with men. AB - The presence of transmission clusters and their relationship to the recruitment chain were investigated in an HIV prevalence assessment survey using respondent driven sampling among men who had sex with men (MSM) in Zagreb, Croatia. HIV infection was found in 18 of 360 participants. Five individuals belong to a transmission cluster of MSM infected with phylogenetically related HIV. All were recruited in later waves (fourth to ninth), suggesting that the population is sexually networked. PMID- 20001314 TI - Mode of coreceptor use by R5 HIV type 1 correlates with disease stage: a study of paired plasma and cerebrospinal fluid isolates. AB - Through the use of chimeric CXCR4/CCR5 receptors we have previously shown that CCR5-tropic (R5) HIV-1 isolates acquire a more flexible receptor use over time, and that this links to a reduced viral susceptibility to inhibition by the CCR5 ligand RANTES. These findings may have relevance with regards to the efficacy of antiretroviral compounds that target CCR5/virus interactions. Compartmentalized discrepancies in coreceptor use may occur, which could also affect the efficacy of these compounds at specific anatomical sites, such as within the CNS. In this cross-sectional study we have used wild-type CCR5 and CXCR4 as well as chimeric CXCR4/CCR5 receptors to characterize coreceptor use by paired plasma and cerebrospinal fluid (CSF) isolates from 28 HIV-1-infected individuals. Furthermore, selected R5 isolates, with varying chimeric receptor use, were tested for sensitivity to inhibition by the CCR5 antagonist TAK-779. Discordant CSF/plasma virus coreceptor use was found in 10/28 patients. Low CD4+ T cell counts correlated strongly with a more flexible mode of R5 virus CCR5 usage, as disclosed by an increased ability to utilize chimeric CXCR4/CCR5 receptors, specifically receptor FC-2. Importantly, an elevated ability to utilize chimeric receptors correlated with a reduced susceptibility to inhibition by TAK-779. Our findings show that a discordant CSF and plasma virus coreceptor use is not uncommon. Furthermore, we provide support for an emerging paradigm, where the acquisition of a more flexible mode of CCR5 usage is a key event in R5 virus pathogenesis. This may, in turn, negatively impact the efficacy of CCR5 antagonist treatment in late stage HIV-1 disease. PMID- 20001316 TI - HIV type 1 in Fiji is caused by subtypes C and B. AB - The HIV epidemic in Fiji remains largely uncharacterized. By February 2009, there were 294 confirmed cases; the majority occurred among the 20- to 39-year old age group and resulted from heterosexual contact. There are currently no published data concerning HIV subtypes in Fiji. In this study, venous blood samples were collected as dried blood spots from 35 HIV-positive individuals in Fiji. HIV-1 subtype was determined for 27 (77%) samples and the presence of four different subtypes, with multiple introductions of two, was demonstrated. Subtype distribution was as follows: 16 (59%) were subtype C, 9 (33%) were subtype B, 1 (4%) was subtype A, and 1 (4%) was subtype G. Phylogenetic analysis showed a clear segregation of the Fijian subtype C isolates and previously published Papua New Guinea subtype C isolates as well as multiple introductions of subtype B. These findings represent the first HIV-1 subtype data from the Fiji Islands. PMID- 20001318 TI - Acute low back pain with radiculopathy: a double-blind, randomized, placebo controlled study. AB - OBJECTIVE: The aim of this study was to investigate the clinical effects of low level laser therapy (LLLT) in patients with acute low back pain (LBP) with radiculopathy. BACKGROUND DATA: Acute LBP with radiculopathy is associated with pain and disability and the important pathogenic role of inflammation. LLLT has shown significant anti-inflammatory effects in many studies. MATERIALS AND METHODS: A randomized, double-blind, placebo-controlled trial was performed on 546 patients. Group A (182 patients) was treated with nimesulide 200 mg/day and additionally with active LLLT; group B (182 patients) was treated only with nimesulide; and group C (182 patients) was treated with nimesulide and placebo LLLT. LLLT was applied behind the involved spine segment using a stationary skin contact method. Patients were treated 5 times weekly, for a total of 15 treatments, with the following parameters: wavelength 904 nm; frequency 5000 Hz; 100-mW average diode power; power density of 20 mW/cm(2) and dose of 3 J/cm(2); treatment time 150 sec at whole doses of 12 J/cm(2). The outcomes were pain intensity measured with a visual analog scale (VAS); lumbar movement, with a modified Schober test; pain disability, with Oswestry disability score; and quality of life, with a 12-item short-form health survey questionnaire (SF-12). Subjects were evaluated before and after treatment. Statistical analyses were done with SPSS 11.5. RESULTS: Statistically significant differences were found in all outcomes measured (p < 0.001), but were larger in group A than in B (p < 0.0005) and C (p < 0.0005). The results in group C were better than in group B (p < 0.0005). CONCLUSIONS: The results of this study show better improvement in acute LBP treated with LLLT used as additional therapy. PMID- 20001317 TI - Palmitic Acid Is a Novel CD4 Fusion Inhibitor That Blocks HIV Entry and Infection. AB - The high rate of HIV-1 mutation and the frequent sexual transmission highlight the need for novel therapeutic modalities with broad activity against both CXCR4 (X4) and CCR5 (R5)-tropic viruses. We investigated a large number of natural products, and from Sargassum fusiforme we isolated and identified palmitic acid (PA) as a natural small bioactive molecule with activity against HIV-1 infection. Treatment with 100 microM PA inhibited both X4 and R5 independent infection in the T cell line up to 70%. Treatment with 22 microM PA inhibited X4 infection in primary peripheral blood lymphocytes (PBL) up to 95% and 100 microM PA inhibited R5 infection in primary macrophages by over 90%. Inhibition of infection was concentration dependent, and cell viability for all treatments tested remained above 80%, similar to treatment with 10(-6)M nucleoside analogue 2', 3' dideoxycytidine (ddC). Micromolar PA concentrations also inhibited cell-to-cell fusion and specific virus-to-cell fusion up to 62%. PA treatment did not result in internalization of the cell surface CD4 receptor or lipid raft disruption, and it did not inhibit intracellular virus replication. PA directly inhibited gp120 CD4 complex formation in a dose-dependent manner. We used fluorescence spectroscopy to determine that PA binds to the CD4 receptor with K(d) approximately 1.5 +/- 0.2 microM, and we used one-dimensional saturation transfer difference NMR (STD-NMR) to determined that the PA binding epitope for CD4 consists of the hydrophobic methyl and methelene groups located away from the PA carboxyl terminal, which blocks efficient gp120-CD4 attachment. These findings introduce a novel class of antiviral compound that binds directly to the CD4 receptor, blocking HIV-1 entry and infection. Understanding the structure affinity relationship (SAR) between PA and CD4 should lead to the development of PA analogs with greater potency against HIV-1 entry. PMID- 20001319 TI - Efficacy of topical aminolevulinic acid photodynamic therapy for the treatment of verruca planae. AB - OBJECTIVE: To evaluate the safety and efficacy of topical aminolevulinic acid (ALA) photodynamic therapy (PDT) in the treatment of verruca planae. METHODS: A total of 18 patients with facial verruca planae was treated with ALA-PDT. A 10% ALA emulsion was applied on facial lesions for a 4-h incubation period after azone infiltrating for 30 min. Skin lesions of verruca planae were irradiated with 120 J/cm(2) using a 635-nm laser for 10 min. Each patient received at least two sessions at intervals of 15 days. Those with residual lesions after the first session were treated with a third PDT. Before treatment and at each return visit, the lesions were photographed. Patient satisfaction was investigated and therapeutic effect was evaluated at 6 months after termination of the therapy. RESULTS: Fifteen days after the first topical ALA-PDT, 10 patients achieved complete response and 7 patients showed improvement. Complete clinical response was observed in 17 patients after two sessions. At 6 months after termination, only one case relapsed among the complete clinical response patients, and the patient satisfaction rate was 88.89%. CONCLUSION: PDT, utilizing the topical application of 10% ALA, has a good result and excellent cosmetic effects for the treatment of verruca planae. PMID- 20001320 TI - Comparison of microleakage of three adhesive systems in class V composite restorations prepared with Er,Cr:YSGG laser. AB - OBJECTIVE: This study compared microleakage of three adhesives in class V (CI V) composite restorations prepared with Er,Cr:YSGG laser. BACKGROUND DATA: Changes produced on enamel and dentin surfaces subsequent to laser irradiation influence microleakage of composite restorations. MATERIALS AND METHODS: CI V cavities were prepared in 80 sound extracted human premolars with Er,Cr:YSGG laser. The teeth were randomly divided into 4 groups (n = 20). In group 1, a total-etch adhesive was used, and two-step and one-step self-etch adhesives were used in groups 2 and 3, respectively. Group 4 was the control group with no adhesives. Following restoration and thermocycling, microleakage was evaluated. To compare microleakage in occlusal and gingival margins in each group, the Wilcoxon test was used. Kruskal-Wallis analysis was used for comparison of occlusal and gingival microleakage, and two-by-two comparisons were made using the Mann Whitney U test. RESULTS: There were significant differences in microleakage between occlusal and gingival margins in each group except for the control group. Significant differences were observed in occlusal and gingival microleakage among all the groups. Two-by-two comparisons revealed significant differences in occlusal microleakage between the control group and the other groups. There were significant differences in gingival microleakage between groups 1 and 2 and groups 1 and 3; however, differences in gingival microleakage between groups 2 and 3 were not significant. CONCLUSION: With all the adhesives, microleakage was significantly higher in gingival margins compared with occlusal margins. In occlusal margins, there were no significant differences in microleakage among the three adhesives. In gingival margins, self-etch adhesives revealed less microleakage than the total-etch adhesive. PMID- 20001321 TI - Effect of LED phototherapy of three distinct wavelengths on fibroblasts on wound healing: a histological study in a rodent model. AB - AIM: The aim of the present investigation was to evaluate histologically fibroblastic proliferation on dorsal cutaneous wounds in a rodent model treated or not with light-emitting diodes (LEDs) of three wavelengths. BACKGROUND: Fibroblasts secrete substances essential for wound healing. There are few reports of LED phototherapy on fibroblast proliferation, mainly in vivo. ANIMALS AND METHODS: Following approval by the Animal Experimentation Committee of the School of Dentistry of the Federal University of Bahia, we obtained 16 young adult male Wistar rats weighing between 200 and 250 g. Under general anesthesia, one excisional wound was created on the dorsum of each animal; they were then randomly distributed into four groups of four animals each: G0, untreated control; G1, red LED (700 +/- 20 nm, 15 mW, 10 J/cm(2)); G2, green LED (530 +/- 20 nm, 8 mW, 10 J/cm(2)); and G3, blue LED (460 +/- 20 nm, 22 mW, 10 J/cm(2)). The irradiation started immediately after surgery and was repeated every other day for 7 days. Animals were killed 8 days after surgery. The specimens were removed, routinely processed to wax, cut, and stained with hematoxylin/eosin (HE). Fibroblasts were scored by measuring the percentage of these cells occupying the area corresponding to wound healing on stained sections. RESULTS: The quantitative results showed that red LED (700 +/- 20 nm) and green LED (530 +/- 20 nm) showed a significant increase in fibroblast numbers (p < 0.01 and p = 0.02) when compared with the control group. CONCLUSION: The use of green and red LED light is effective in increasing fibroblastic proliferation on rodents. PMID- 20001322 TI - Production of reactive oxygen species from photosensitizers activated with visible light sources available in dental offices. AB - OBJECTIVES: The aim of this study was to assess the ability of commonly available red- or blue-light dental sources to generate reactive oxygen species (ROS) from photosensitive chemicals that might be useful for photodynamic antimicrobial chemotherapy (PACT). BACKGROUND: Although the use of red diode lasers is well documented, there is limited information on how useful blue-light sources might be for PACT in dental contexts. MATERIALS AND METHODS: A diode laser (Periowave; see Table 1 for material and equipment sources) emitting red light (660-675 nm) was used to activate toluidine blue; riboflavin and pheophorbide-a polylysine (pheophorbide-a-PLL) were photoactivated using an Optilux 501 curing unit emitting blue light (380-500 nm). Ozone gas (generated by OzoTop, Tip Top Tips, Rolle, Switzerland), sodium hypochlorite, and hydrogen peroxide were used for comparison. ROS production was estimated using an iodine-triiodide colorimetric assay, and ROS levels were plotted versus concentration of chemicals to determine each chemical's efficiency in ROS production. One-way ANOVA with Tukey post hoc analysis (alpha = 0.05) was used to compare the efficiencies of ROS production for the various chemicals. RESULTS: Sodium hypochlorite, hydrogen peroxide, and ozone gas produced ROS spontaneously, whereas pheophorbide-a-PLL, riboflavin, and toluidine blue required light exposure. The efficiency of ROS production was higher for pheophorbide-a-PLL and toluidine blue than for ozone gas or riboflavin (p < 0.05). Hydrogen peroxide was the least efficient ROS producer. CONCLUSIONS: The results of the current study support the use of blue- or red-light-absorbing photosensitizers as candidates to produce ROS for clinical applications. Blue light photosensitizers were as efficient as red-light photosensitizers in producing ROS and more efficient than the oxidant chemicals currently used for dental disinfection. PMID- 20001323 TI - Salmonella spp. infection in imported 1-day-old chicks, ducklings, and turkey poults: a public health risk. AB - The occurrence of Salmonella in 750 birds was assessed. The samples included the internal organs (caecal pouches, yolk sac, liver, and lung) of imported 1-day-old chicks (n = 150), grandparent chicks (n = 150), breeder chicks (n = 150), ducklings (n = 150), and turkey poults (n = 150), and paper-lined boxes (n = 250). Salmonellae isolated from the internal organs and paper-lined box of 1-day old chicks, ducklings, and poults were mostly evident from the paper-lined box followed by caecal samples. Imported 1-day-old grandparent flocks were Salmonella free. Although 23.3% of the imported breeder flocks were positive for Salmonella, the imported duckling flocks and day-old turkey poults exhibited 19.3% and 12.6%, respectively. The widest diversity in isolated salmonellae was from the 1-day-old chicks where Salmonella Newport, Salmonella Kentucky, Salmonella Enteritidis, Salmonella Shubra, Salmonella Saintpaul, and Salmonella Agona were isolated. On the other hand, two Salmonella serovars were isolated from the imported breeders, Salmonella Shubra and Salmonella Shipley, and from the imported ducklings, Salmonella Shubra and Salmonella Saintpaul. The three Salmonella serovars isolated from the imported day-old turkey poults were Salmonella Shubra, Salmonella Newport, and Salmonella Saintpaul. The high percentage and diversity of Salmonella isolation from the imported birds cause concern because of the zoonotic potential of this agent and its economical importance to the local commercial poultry breeding industry. From 80 samples investigated for Salmonella, the positivity of the standard microbiological technique method was 17.5% and of the polymerase chain reaction method (Salmonella-specific invA gene) was 22.5%. The concordance between the two methods was 90% (k = 0.850). Our results indicated that the polymerase chain reaction approach is better than culturing for detecting Salmonella in poultry samples when using the preenriched medium combinations used in this study. PMID- 20001324 TI - An improved quarantine method for mangoes against the Mexican fruit fly based on high-pressure processing combined with heat. AB - The Mexican fruit fly Anastrepha ludens Loew (Diptera: Tephritidae) is one of the most important insects infesting mangoes, citrus, and other fruits in Mexico and other Latin-American countries. Quarantine methods approved to destroy this insect decrease the shelf life of commodities. The objective of this study was to determine the effect of high-pressure processing using an initial temperature of 50 degrees C on the survivorship of eggs and larvae of the Mexican fruit fly. Eggs and larvae were pressurized at 25, 50, 75, 100, or 150 MPa for 0, 5, 10, or 20 min. The hatching ability of pressurized eggs of 1, 2, 3, and 4 days old and survivorship of the first, second, and third instars were registered. Further, the ability to pupate was studied in surviving third instars. The results showed that eggs were more resistant than larvae to the high-pressure processing. Treatments at 150 MPa at initial 50 degrees C for 10 min destroyed all eggs and larvae of A. ludens, indicating that this process might be useful as a quarantine method for infested mangoes or other fruits. PMID- 20001325 TI - Antibacterial activity of Ecklonia cava against methicillin-resistant Staphylococcus aureus and Salmonella spp. AB - Ecklonia cava is a brown alga (Laminariales, Phaeophyta) growing on the subtidal rocky shores of Korea. It has antioxidant, antidiarrhea, and anticoagulant effects. In this study, the antimicrobial activity of E. cava EtOH extract and its fractions (n-hexane, CH2Cl2, EtOAc, n-BuOH, and H2O) were investigated against methicillin-resistant Staphylococcus aureus and Salmonella spp. The E. cava EtOAc fraction showed good antibacterial activity against all bacteria. Eckol isolated from E. cava EtOAc fraction showed antimicrobial activity against all the tested strains. The minimum inhibitory concentration of eckol against S. aureus strains ranged from 125 to 250 microg/mL and 125 to 250 microg/mL for Salmonella strains. The fraction inhibitory concentration index of eckol in combination with ampicillin ranged from 0.31 to 0.5, indicating remarkable synergism against S. aureus. However, against Salmonella gallinarum ATCC 9184 and Salmonella typhimurium, it ranges from 0.75 to 1.0. The combinations of eckol + ampicillin exhibited improved inhibition of S. aureus and Salmonella with synergy or additive effect. We suggest that eckol ingredients of the E. cava against S. aureus and Salmonella have antibacterial activity. PMID- 20001326 TI - Molecular and pathogenic characterization of Salmonella enterica serovar Bovismorbificans strains of animal, environmental, food, and human origin in Hungary. AB - In this study, we characterized 110 strains of Salmonella enterica serovar Bovismorbificans contaminating environment, animals, food of animal origin, and human, to assess their significance along the food chain in Hungary. Additionally, five strains from Germany were tested for comparative purposes. Characterization involved antibiotic susceptibility testing, class 1 integron detection by polymerase chain reaction, plasmid profiling, virulotyping (using virulence gene-specific polymerase chain reactions), and pulsed-field gel electrophoresis. Pathogenic potential of selected strains was tested in orally infected 1-day-old specific pathogen-free chicks. Eighty-two percent of the strains were susceptible to the 16 antibiotics tested, and none of them had class 1 integron. A multidrug-resistant human isolate harbored a bla(SHV5)-type extended-spectrum beta-lactamase gene, first reported in this serotype. All the strains possessed avrA, ssaQ, mgtC, spi4, and sopB genes indicating the presence of Salmonella pathogenicity islands 1-5, respectively, missed the phage-related genes sopE and gipA, but retained the phage-related gene sodC1. An approximately 90 kb large plasmid was characteristic to 80% of the strains, all of which carried the spvC gene. In vivo colonization testing of four selected strains in 1 day-old chicks resulted in significantly reduced liver and spleen colonization ability as compared with the Salmonella Enteritidis control strain, whereas their caecal colonization ability differed less from that of Salmonella Enteritidis. Pulsed-field gel electrophoresis data revealed the dominance of two pulsotypes (C2 and C5) without any specific temporal, geographical, and/or source-related linkages. The results show that Salmonella Bovismorbificans studied here are less invasive than Salmonella Enteritidis, but they may colonize and persist in several animal species and successfully contaminate meat products of different animal origin in Hungary. PMID- 20001327 TI - The combination of energy-dependent internal adaptation mechanisms and external factors enables Listeria monocytogenes to express a strong starvation survival response during multiple-nutrient starvation. AB - The goal of this study was to characterize the starvation survival response (SSR) of a wild-type Listeria monocytogenes 10403S and an isogenic DeltasigB mutant strain during multiple-nutrient starvation conditions over 28 days. This study examined the effects of inhibitors of protein synthesis, the proton motive force, substrate level phosphorylation, and oxidative phosphorylation on the SSR of L. monocytogenes 10403S and a DeltasigB mutant during multiple-nutrient starvation. The effects of starvation buffer changes on viability were also examined. During multiple-nutrient starvation, both strains expressed a strong SSR, suggesting that L. monocytogenes possesses SigB-independent mechanism(s) for survival during multiple-nutrient starvation. Neither strain was able to express an SSR following starvation buffer changes, indicating that the nutrients/factors present in the starvation buffer could be a source of energy for cell maintenance and survival. Neither the wild-type nor the DeltasigB mutant strain was able to elicit an SSR when exposed to the protein synthesis inhibitor chloramphenicol within the first 4 h of starvation. However, both strains expressed an SSR when exposed to chloramphenicol after 6 h or more of starvation, suggesting that the majority of proteins required to elicit an effective SSR in L. monocytogenes are likely produced somewhere between 4 and 6 h of starvation. The varying SSRs of both strains to the different metabolic inhibitors under aerobic or anaerobic conditions suggested that (1) energy derived from the proton motive force is important for an effective SSR, (2) L. monocytogenes utilizes an anaerobic electron transport during multiple-nutrient starvation conditions, and (3) the glycolytic pathway is an important energy source during multiple-nutrient starvation when oxygen is available, and less important under anaerobic conditions. Collectively, the data suggest that the combination of energy dependent internal adaptation mechanisms of cells and external nutrients/factors enables L. monocytogenes to express a strong SSR. PMID- 20001328 TI - Genetic similarity of Campylobacter isolates in humans, food, and water sources in central Poland. AB - Campylobacter spp. is an important cause of gastroenteritis in humans throughout the world. However, sources of these infections are often difficult to identify. Therefore, this study aimed at analyzing the genetic relatedness of Campylobacter isolates from environmental and food samples as well as stool specimens of diarrheal patients obtained in a single geographic region in Poland. Only a few Campylobacter jejuni isolates (4/18, 22%) could be assigned to one cluster, whereas the majority were unrelated. In contrast, the majority of Campylobacter coli strains (25/35, 71%) belonged to three pulsed-field gel electrophoresis (PFGE) clusters containing isolates of various origins (i.e., water samples, chicken carcasses, and humans). Isolates belonging to the clusters showed also similar antibiotic resistance patterns and similar genotypes with respect to the occurrence of the virB11 and iam genes. This suggests that Campylobacter strains may circulate between humans, poultry, and recreational water sources in the rural region in central Poland. PMID- 20001329 TI - A pilot study of the microbiological quality of culturally diverse, ready-to-eat foods from selected retail establishments in Melbourne, Australia. AB - In recent years, there has been an increasing number of foodborne outbreaks linked to the consumption of culturally diverse foods. This appears to be because of the increasing quantity of culturally diverse foods available and a preference to store these foods, some of which are considered potentially hazardous, at ambient temperature. This practice may contravene temperature requirements defined by the Food Standards Code. A lack of understanding of the hazardous nature of some culturally prepared foods also poses difficulties in applying the Australian food safety legislation by regulators. This pilot study examined the normal microbiota of four culturally diverse foods: nem chua, che dau trang, kueh talam, and banh tet nhan man, which are traditionally stored and consumed at ambient temperature. Challenge testing was conducted to investigate the ability of these foods to support the growth of foodborne bacterial pathogens. Two of the products (kueh talam and che dau) were found to be microbiologically unsatisfactory because of the high standard plate counts. Challenge testing indicated that kueh talam, che dau, and banh tet nhan man were able to support the growth of Bacillus cereus, Escherichia coli, Staphylococcus aureus, and Salmonella (1-2 log increases over 6 hours at 25 degrees C), suggesting that these foods may require temperature control during storage. However, nem chua was unable to support the growth of test bacteria, probably because of its acidic nature (pH 4.5), suggesting that ambient storage of this food may be safe. This study provided some preliminary evidence to support the need for further sampling and challenge testing of these products. PMID- 20001330 TI - A simple method for estimating the optimum number of prostate biopsy cores needed to maintain high cancer detection rates while minimizing unnecessary biopsy sampling. AB - OBJECTIVE: The objective was to provide urologists with a simple basis for optimizing the number of prostate biopsy cores that should be taken. METHODS: The records of 1024 patients who had undergone transrectal ultrasound-guided biopsies were reviewed. The prostate volume was divided by the number of biopsies to obtain the volume/biopsy ratio (VBR). Univariate and multivariate analyses were performed to determine the best predictors for positive biopsies. RESULTS: The analysis included 939 patients who had prostatic-specific antigen <20 ng/mL. The significant independent variables for positive biopsies were age, prostatic specific antigen, and prostate volume and VBR (p < 0.001). VBR had the strongest correlation coefficient out of all significant variables. Stepwise analysis showed a consistent increase in cancer detection rates as VBR was decreased. The detection rates for VBRs of 2, 3, and 4 were 59%, 53%, and 50%, respectively. The detection rates dropped sharply to 42% and 30% for VBRs of 5 and 6, respectively. Cancers diagnosed with low VBRs were similar to those diagnosed with high VBRs in regard to Gleason scores and percentages of cancer in the prostatectomy specimens. CONCLUSION: Using VBR of 4 maintains high cancer detection rates without taking an excessive number of biopsy specimens. This is a simple and easy to-remember method. PMID- 20001331 TI - Effect of mobile unidirectional air flow unit on microbial contamination of air in standard urologic procedures. AB - BACKGROUND: Infection is one of the most feared complications of surgery. New instrumentation is being developed to reduce deposition of bacteria. METHODS: We investigated 45 major surgical procedures (21 radical nephrectomies [RN] and 24 radical retropubic prostatectomies [RRP]) in our urology department during 2007. In about one-half of the interventions, an ultraclean air flow mobile (UAF) unit was used. Bacterial sedimentation was evaluated by nitrocellulose membranes placed on the instrument tray and by settle plates positioned at four points in the operating room. In 27 operations, an additional membrane was located near the incision. RESULTS: Bacterial counts on the nitrocellulose membranes during RN were 230 colony-forming units (cfu)/m(2)/h with the UAF unit and 2,254 cfu/m(2)/h without the unit (p = 0.001). During RRP, the values were 288 cfu/m(2)/h and 3,126 cfu/m(2)/h respectively (p = 0.001). The membrane placed near the incision during RN showed a microbial count of 1,235 cfu/m(2)/h with the UAF unit and 5,093 cfu/m(2)/h without the unit (p = 0.002); during RRP, the values were 1,845 cfu/m(2)/h and 3,790 cfu/m(2)/h, respectively (difference not significant). Bacterial contamination detected by settle plates during RN showed a mean value of 2,273 cfu/m(2)/h when the UAF unit was used and 2,054 cfu/m(2)/h without the unit; during RRP, the values were 2,332 cfu/m(2)/h and 2,629 cfu/m(2)/h with and without the UAF unit, respectively (NS). No statistically significant differences were detected in the clinical data registered in patients operated on under standard conditions and while the UAF unit was functioning. CONCLUSIONS: The UAF appears able to reduce microbial contamination at the operating table, reaching a bacterial number obtained in ultraclean operating theatres. PMID- 20001332 TI - Predicting death in necrotizing soft tissue infections: a clinical score. AB - BACKGROUND: Necrotizing soft tissue infections (NSTIs) are associated with a high mortality rate; however, there is no uniform way to categorize the severity of this disease early in its course. The goal of this study was to develop a clinical score based on data available at the time of initial assessment to aid in stratifying patients according to their risk of death. METHODS: A cohort of all 350 patients admitted with NSTI to two institutions over a nine-year period was examined retrospectively. Using random split sampling, two datasets were created: Prediction (PD) and validation (VD). Multivariable stepwise regression analysis of the PD identified independent predictors of death using data available at the time of admission. Model performance was evaluated for accuracy, discrimination, and calibration. A clinical score to predict death was created, and using the Trauma and Injury Severity Score (TRISS) methodology, the score was validated on the VD (z-statistic). RESULTS: Six admission parameters independently predicted death: Age > 50 years, heart rate > 110 beats/min, temperature <36 degrees C, white blood cell count > 40,000/mcL, serum creatinine concentration > 1.5 mg/dL, and hematocrit > 50%. The accuracy of this model was 86.8%; the area under the receiver-operating characteristic curve was 0.81, and the Hosmer-Lemeshow statistic was 11.8. Additionally, the score had excellent performance in evaluation on the VD (z-score/statistic 0.23 to - 0.83). CONCLUSION: A clinical score that categorizes patients with NSTI according to the risk of death was created. It uses simple variables, all available at the time of first assessment. It stratifies patients according to disease severity and can guide the use of aggressive or novel therapeutic strategies and selection of patients for clinical trials. PMID- 20001333 TI - Implementation of tight glucose control for critically ill surgical patients: a process improvement analysis. AB - BACKGROUND: Tight glucose control has been advocated as a method to improve outcomes of surgical critical care. However, continuous infusion of insulin has potential morbidity (e.g., neurologic consequences of hypoglycemia), and it remains unclear to what degree the glucose concentration must be controlled. We examined our performance in instituting a protocol for tight glucose control in our surgical intensive care unit (ICU). METHODS: Prospective study of 220 consecutive patients (February, 2003-March, 2006) who received an infusion of insulin for glucose control for >24 h by protocol. Data collected included age, acuity (Acute Physiology and Chronic Health Evaluation [APACHE] III) score, sex, history of diabetes mellitus, organ dysfunction (Marshall), and death or survival. Infusion-related data included initial glucose concentration, time to glucose <120 mg/dL, h/day of glucose <110 mg/dL and <140 mg/dL, duration of infusion (days), insulin units/day, year of therapy, and complications. Analysis was performed by chi(2), analysis of variance, and logistic regression, with p < 0.05 considered significant. RESULTS: Insulin drips were required by 10.2% of patients (287/2,804); 29 of these (10.1%) had diabetes mellitus. The mean APACHE III score for the treated patients was 77 +/- 2 (standard deviation), and the mortality rate was 24%. Hypoglycemia (<60 mg/dL) occurred in 4.2% of patients. The trigger insulin concentration decreased over time (2003 vs. 2005) from 249 +/ 14 to 160 +/- 5 mg/dL, and the h/day of glucose <140 increased from 11 +/- 1 to 16 +/- 1. However, age, acuity, APACHE III, days of insulin, time to achieve glucose <120, h/day of glucose <110, and mortality rate were unchanged. By logistic regression, only the year of treatment (odds ratio [OR] 1.871; 95% confidence interval [CI] 1.177, 2.972; p = 0.008] predicted success in controlling the blood glucose concentration to <140 mg/dL; age, illness severity, diabetes history, and trigger glucose concentration [OR 0.996; 95% CI 0.992, 1.001; p = 0.11] did not. CONCLUSIONS: Success in implementing tight glucose control was modest, albeit improving, despite a specific protocol for administration. No medical reason could be identified for inability to achieve tight glucose control; therefore, successful implementation must be volitional. Education, particularly regarding hypoglycemia, and possible refinement of our protocol may improve our ability to control blood glucose in our ICU. PMID- 20001334 TI - Course in scientific writing--the Arctic experience. PMID- 20001335 TI - Validation of the Hemo_Control instrument for determination of B-haemoglobin concentration in primary health care. AB - OBJECTIVE: The aim of this study was to evaluate the analytical performance of a new portable haemoglobinometer, Hemo_Control (EKF-diagnostic, GmbH, Germany), which measures haemoglobin concentration in venous and capillary samples. METHOD: The within series and between series imprecision of the Hemo_Control instrument were calculated after measuring the concentration of venous samples under standardized conditions; by experienced laboratory technicians in a hospital laboratory, and venous and capillary samples under conditions similar to where the instrument is intended for use; by personnel at two primary health care centres. The bias of the Hemo_Control instrument was calculated as the difference between its results and results obtained with a Coulter LH 750 instrument traceable to the ICSH reference method. RESULTS: The uncertainty of the Hemo_Control instrument for venous samples was lower than the quality goal of +/- 5% considered acceptable for patient care. High within series imprecision (5.5%) was observed for measurements of capillary blood samples in one of the primary care centres, whereas adequate analytical performance was obtained at the other centre. The Hemo_Control instrument showed negligible bias of +0.8 g/L for both venous and capillary samples in primary health care. CONCLUSION: The observed uncertainty indicates that Hemo_Control is appropriate for near patient testing using venous samples. Capillary samples may be used if sampling skills are adequate. PMID- 20001336 TI - Resorption and elimination kinetics of benzoic acid during chemical necrectomy in deep burns. AB - OBJECTIVE: Chemical necrectomy is an alternative to the surgical or sharp necrectomy for the removal of necrotic parts of the skin in the treatment of deep burns. The aim of our work was to monitor the dynamics of resorption and elimination of benzoic acid applied to the burnt skin. METHODS: The set consisted of 10 patients (9 men; 1 woman) aged 25-57 years with IIb-III-degree skin burns. 40% benzoic acid in white petrolatum was applied to the burnt area to the extent of 3-5% of TBSA (total body surface area) for a period of 48 hours. The concentrations of benzoic acid, hippuric acid, and glycine in the serum was monitored at the 10th, 20th, 60th, 120th, 240 th and 360 th minute thereafter and further at the 12th, 24th, 48th, and 72nd hour; the excretion of hippuric acid in urine was monitored in six 12-hour intervals. RESULTS: The highest concentration of benzoic acid in the serum was detected in the 60th minute sample (0.094+/ 0.074 mmol/L) and of hippuric acid in the 120th minute sample (0.234+/-0.088 mmol/L) from the application of benzoic acid to the burnt skin. In the period between the 6th and 48th hour, the average concentration of benzoic acid in the serum ranged between 0.042 and 0.03 mmol/L. In this period there was also a significant decrease in serum glycine concentration (p<0.05). During the 48-hour application of benzoic acid to the burnt skin, 46.0-145 mmol of hippuric acid was excreted in urine. CONCLUSION: Chemical necrectomy with the use of 40% benzoic acid led only to a moderate increase of its concentration in the serum. After its resorption from the wound area it is transformed to hippuric acid, which is promptly excreted in urine. PMID- 20001337 TI - High-density lipoprotein is superior to B-type natriuretic peptide as a marker of systolic dysfunction in an elderly general population. AB - OBJECTIVES: Natriuretic peptides are well-known indicators of left ventricular dysfunction. The aim of this study was to examine whether the levels of B-type natriuretic peptide (BNP) and HDL are associated with systolic dysfunction in an elderly general population. DESIGN: The 355 study subjects aged 75 years or more were studied in the City of Kuopio, in East Finland. The association between clinical variables as well as echocardiographic left ventricular ejection fraction and BNP, total cholesterol and HDL were investigated using logistic and linear regression analysis. RESULTS: When tested as a dichotomous variable (EF > or T /c.460dupT) was found in the proband. The paternal c.211C > T is a novel point mutation, resulting in an immediate stop codon (p.Arg71X). The maternal c.460dupT is a frameshift mutation introducing an earlier stop codon (p.Phe156AspfsX10). Both mutations could be expected to lead to truncated protein of podocin. Abnormal expression and distribution of the mutated protein were also exhibited in the patient. CONCLUSIONS: The compound heterozygous mutation in NPHS2 may explain the development of SRNS in this family. p.Arg71X is a novel disease causing mutation leading to a deficient expression of podocin. PMID- 20001347 TI - Current clinical and laboratory practice for the investigation of the antiphospholipid syndrome: findings from the 2008 Australasian antiphospholipid antibody survey. AB - BACKGROUND: The antiphospholipid syndrome (APS) is an autoimmune condition characterised by vascular thromboses and/or pregnancy morbidity, and its diagnosis currently requires laboratory evidence for the presence of antiphospholipid antibodies (aPL). aPL are identified using a large number of laboratory procedures based on one of two distinct test processes, namely 'solid' or 'liquid' phase assays. The former include anticardiolipin antibodies (aCL) and anti-beta-2-glycoprotein-I antibodies (aB2GPI), while the latter are centred on clot-based tests used to identify the lupus anticoagulant (LA). Depending on their clinical presentation, affected individuals might be seen by a variety of clinical specialties including general physicians and general practionners, with a potentially wide variation in the aPL assays requested. METHODS: The current report summarises findings from the '2008 Australasian antiphospholipid antibody survey', a simple 5-step survey that assessed current clinical and laboratory practice in the investigation of APS. The survey was despatched via various clinical and scientific professional bodies. RESULTS: Responses were received from 130 scientific and clinical personnel, primarily haematology based (94/130; 72%) or immunology based (34/130; 26%). Most respondents (97/130; 75%) ordered or recommended tests for solid phase aPL testing, and most also attempted to grade these tests and their isotypes. Most were familiar with aCL and aB2GPI testing, and tended to request primarily IgG and IgM isotypes of these antibodies. Only a small number of respondents requested/recommended IgA isotype testing of these antibodies or the other solid phase aPL assays (e.g., anti-prothrombin). A similar number of respondents (104/130; 80%) also ordered or recommended tests for LA, and most also attempted to grade these tests and their isotypes. Some discipline-related biases were also evident, in that 32/34 (94%) of immunology based respondents identified that they ordered or recommended specific solid phase tests for aPL, whereas only 62/94 (66%) of haematology-based respondents did so. In contrast, 83/94 (88%) of haematology-based respondents identified that they ordered or recommended specific LA test procedures, whereas only 18/34 (53%) of immunology-based respondents did so. CONCLUSION: To our knowledge, this report represents the first ever attempt to survey a wide range of clinical and scientific personnel regarding ordering and recommending practices for aPL testing, and provides a snapshot of current clinical and laboratory practice for the investigation of APS in Australia and New Zealand. Most respondents to our survey still consider the immunoglobulin G (IgG) aCL test to be a useful first line solid phase aPL test, and the dilute Russell's viper venom time (dRVVT) assay to be the most useful LA test. PMID- 20001348 TI - Epidemiological analysis of Streptococcus pyogenes infections in Hong Kong. AB - AIMS: The aim of this study was to characterise clinical and microbiological features of isolates obtained from both invasive and non-invasive Streptococcus pyogenes infections in Hong Kong, between October 2005 and April 2008. METHOD: Clinical data of invasive isolates were collected retrospectively. Altogether 281 isolates were emm sequence typed and tested for antimicrobial susceptibility using disk diffusion method. Detection of the presence of the streptococcal pyrogenic exotoxin genes was also carried out. RESULTS: emm1, emm4 and emm12 were the most prevalent in both the invasive and non-invasive groups with an increase in incidence of emm22 compared with a previous study. emm22 was associated with invasive cellulitis and wound infection. The overall rate of erythromycin resistance was 25.6% and was significantly higher in emm22 strains (85.7%). The phage-encoded superantigen gene speA was exclusively associated with emm1 in both invasive and non-invasive isolates. CONCLUSION: This study revealed a changing epidemiology of S. pyogenes infection in Hong Kong, with a unique pattern compared with other Asian countries. Invasiveness is not related to the presence of speA, speC or ssa genes and the antimicrobial resistance rate was high for macrolides. The findings have an implication on the use and efficacy of the polyvalent S. pyogenes vaccine under development. PMID- 20001349 TI - Norovirus Ridaquick: a new test for rapid diagnosis of norovirus. PMID- 20001350 TI - Analytical interference of carbamazepine on the Abbott TDx and Abbott Axsym tricyclic antidepressant assays. PMID- 20001351 TI - Antimicrobial action of atorvastatin and rosuvastatin. PMID- 20001352 TI - Superficial myofibroblastoma of the lower female genital tract in the uterine cervix showing focal pseudosarcomatous morphology. PMID- 20001353 TI - Endometrial adenocarcinoma in spontaneous abortion. PMID- 20001354 TI - Mediastinal seminoma with multiple KIT gene mutations. PMID- 20001355 TI - Plexiform angiomyxoid myofibroblastic tumour of the stomach. PMID- 20001356 TI - Collision tumour: atypical fibroxanthoma and invasive melanoma. PMID- 20001357 TI - Thymic marginal zone B-cell lymphoma of mucosa-associated lymphoid tissue-type in a patient with Sjogren's syndrome and cryoglobulinaemia. PMID- 20001358 TI - Ocular mantle cell lymphoma with aberrant CD10 expression: a potential diagnostic pitfall. PMID- 20001359 TI - Cardiac lymphoma presenting as concentric pseudo-hypertrophic cardiomyopathy in a previously unrecognised HIV patient. PMID- 20001360 TI - A mitotically active, intramuscular glomus tumour with no recurrence at 12 months. PMID- 20001363 TI - Psychiatry: responding to population health needs. PMID- 20001362 TI - Adolescent toluene exposure produces enduring social and cognitive deficits in mice: an animal model of solvent-induced psychosis. AB - OBJECTIVES: Abuse of toluene-containing volatile solvents by adolescents is a significant public health problem. The present study characterized the long-term behavioural and neurochemical consequences of toluene exposure during adolescence. METHODS: Male NMRI mice received one injection per day of either toluene (600 mg/kg) or corn oil during postnatal days (PN) 35-37 and (750 mg/kg) during PN38-39 and PN42-46. A variety of psychiatric disorder-relevant behavioural tests were examined at PN56-P84. RESULTS: The toluene-exposed mice were significantly deficient in the social interaction test, nesting behaviour, social dominance tube test, and novel objective recognition test. However, toluene exposure did not affect locomotor activity and behavioural profiles in the forced swimming test, tail suspension test, emergence test and elevated plus maze. Neurochemically, the turnover rates of dopamine in the prefrontal cortex, striatum and nucleus accumbens were reduced in toluene-treated mice. CONCLUSIONS: Adolescent toluene exposure leads to social deficits and cognitive impairment at adulthood as well as neurochemical dysfunction in mice, which correlate with the symptoms observed in patients suffering from solvent-induced psychosis. These findings highlight the need for understanding the effects of solvent abuse on the developing nervous system and reveal an animal model suitable for research into pathophysiology of neurological and psychiatric consequences of solvent abuse. PMID- 20001364 TI - 'Radical hope' and rain: climate change and the mental health of Indigenous residents of northern Australia. AB - Objective: This paper considers the short, intermediate and longer term effects of climate change in relation to the mental health of Indigenous residents of northern Australia, and what these effects mean in terms of supporting adaptation and resilience. Conclusions: Indigenous Australians have contended with change for millennia, with the drivers shifting from ecological to social pressures since European colonization. Climate change resulting from human activities introduces a new set of change forces which will in the short term be mediated by economic and social effects internationally. PMID- 20001365 TI - Pearl in the oyster: climate change as a mental health opportunity. AB - Objective: Our world faces potentially catastrophic climate change and we have limited capacity to adapt to rapid or extreme climatic changes. As a result, we can expect significant adverse impacts on health. This includes mental health, a major and growing global concern. It is essential to understand how to respond quickly, effectively and within a manageable budget. The aim of this paper is to propose that the adverse consequences of climate change might offer a subtle but important mental health promotion opportunity which meets these criteria. Conclusions: Climate change will affect mental health directly through increasing exposure to trauma, and indirectly through harming physical health and damaging the physical environment on which economic opportunity depends. Disadvantaged people and communities, especially in rural and remote Australia, will be hardest hit. Seminal work by Alexander Leighton demonstrated how profound disadvantage and associated elevated psychiatric morbidity could be addressed by building community capacity. His methods might be adapted, using the need to address adverse climate change as an opportunity to build social capital. Social capital is associated with a wide range of socioeconomic and health advantages, particularly decreased psychiatric morbidity. PMID- 20001366 TI - Comment. PMID- 20001367 TI - Emotion modelling towards affective pathogenesis. AB - Objective: There is a need in psychiatry for models that integrate pathological states with normal systems. The interaction of arousal and emotion is the focus of an exploration of affective pathogenesis. Method: Given that the explicit causes of affective disorder remain nascent, methods of linking emotion and disorder are evaluated. Results: A network model of emotional families is presented, in which emotions exist as quantal gradients. Morbid emotional states are seen as the activation of distal emotion sites. The phenomenology of affective disorders is described with reference to this model. Recourse is made to non-linear dynamic theory. Conclusions: Metaphoric emotion models have face validity and may prove a useful heuristic. PMID- 20001368 TI - Two models of suicide. AB - Objective: The aim of this paper is to present two models of separate but related aspects of suicide, developed with a view to improving understanding and management of this behaviour. Conclusions: First, the predicament model of suicide posits that all suicide represents an escape from a predicament and associated distress. Predicaments are composed of either external (environmental) or internal (mental disorders) factors, or both. Suicide occurs when a threshold is exceeded on a suicide risk ladder, and the degree of movement toward the threshold in response to a particular stressor depends on a range of factors. Second, the suicide pathways model integrates medical and sociological concepts, with distress as the central component, and three run-offs: mental disorder, medicalized and a non-mental disorder (egoistic/anomic; reaction) suicide. PMID- 20001369 TI - Australian young people's awareness of headspace, beyondblue and other mental health organizations. AB - Objective: The aim of this paper is to assess young people's awareness of mental health organizations supporting their age group. Of particular interest was awareness of headspace, which was created in 2006 to provide youth-oriented mental health services, and of beyondblue, which aims to raise community awareness of depression, anxiety and related disorders. Method: A telephone survey was carried out on a national sample of young people who were part of a 2 year follow-up study of youth mental health literacy. Data were analysed for those aged 13-25 years. Results: Awareness was highest for beyondblue and telephone helplines; headspace had established some awareness, particularly in areas where its services operate. However, awareness was unrelated to the participant's psychological symptoms in the past year. Conclusions: If youth oriented services are to be successful, young people need to know about them. Awareness campaigns need to be targeted at the sub-group who have greatest need for these services, namely those with mental health problems. PMID- 20001370 TI - Evaluation of a mental health literacy training program for junior sporting clubs. AB - Objective: The aim of this study was to describe and evaluate a training program designed to improve mental health literacy in junior sporting club coaches and leaders. It was anticipated that participants would demonstrate an improvement in knowledge, confidence, and attitudes in relation to mental disorders and help seeking. Methods: A training program called Read the Play was delivered to 40 participants from junior AFL football and netball leagues in the Barwon region of Victoria. The effects of the training were evaluated using pre- and post questionnaires. Results: The course led to significant improvement in knowledge about mental disorders, increased confidence in helping someone with a mental disorder and more positive attitudes towards people with mental disorders. Conclusions: Training programs delivered within sporting settings may be effective in improving mental health literacy. Future evaluations would benefit from assessing whether these changes are sustained over time and whether trainees subsequently assist young club members to seek appropriate professional help. PMID- 20001371 TI - Description and qualitative evaluation of Jigsaw, an integrated young persons' mental health program. AB - Objective: The aim of this paper is to describe the establishment of an integrated young person's mental health service and the findings of a qualitative evaluation conducted 2 years after its establishment. Method: A qualitative evaluation of the service was undertaken using a semi-structured interview, a service satisfaction survey and partnership analysis tool. Results: The major problems encountered in establishing the service were insufficient recognition of the cultural challenges in working together, difficulty in recruiting general practitioners, establishing a youth friendly environment and maintaining the quality of the relationship between partners. Conclusion: Despite almost 3 years of preparation, many important aspects of change management were underestimated or inadequately attended to. PMID- 20001372 TI - Challenges in psychiatric classification: the case of generalized anxiety disorder. AB - Objective: This paper aims to describe some of the challenges in psychiatric classification, using generalized anxiety disorder (GAD) as an example. A range of different conceptualizations of GAD are presented. Some are based on theoretical formulations, while others draw on epidemiological data. Each has merit, but also deficiencies. The evolution of diagnostic criteria is not simply a theoretical exercise, but reflects assumptions about the nature of the underlying pathology and the relationships between different disorders. Furthermore, these criteria determine which subjects are included in research and in clinical trials, so they shape the further development of psychiatric classification systems. Conclusion: The controversies about the classification of GAD illustrate the complexities and challenges of developing a valid classification system for psychiatric disorders. PMID- 20001373 TI - Shyness 5: the clinical effectiveness of Internet-based clinician-assisted treatment of social phobia. AB - Objective: The present study explores the clinical effectiveness of an Internet based cognitive behavioural treatment program for social phobia (the Shyness program) administered by a psychiatric registrar as part of standard clinical treatment at an outpatient mental health service (the Anxiety Disorders Clinic, St Vincent's Hospital, Sydney). Method: Seventeen individuals with social phobia were assigned to this 8 week clinician-assisted program. All patients had access to online lessons, homework assignments, an online discussion forum with other patients, and regular emails from the clinician. Both completer and intention-to treat models were used in data analyses. Results: Fourteen (82%) patients completed all six lessons within the 8 weeks, and 11/17 (65%) completed post treatment questionnaires. Mean within-group effect sizes (Cohen's d) for the two social phobia measures were 1.06 and 0.77, based on completer and intention-to treat analyses, respectively. Data indicate that the procedure was acceptable to patients. Conclusions: These results indicate that the procedure was clinically effective when operated as part of standard clinical treatment. These results provide further support for the potential clinical utility of the Shyness program. PMID- 20001374 TI - The prediction of imminent aggression in patients with mental illness and/or intellectual disability using the Dynamic Appraisal of Situational Aggression instrument. AB - Objective: The aim of this paper was to determine whether imminent aggression in psychiatric inpatients can be accurately predicted using a structured risk assessment instrument, the Dynamic Appraisal of Situational Aggression (DASA). Methods: This prospective validation study involved 10 013 DASA risk assessments of patients residing in a psychiatric hospital. Twenty-four hours after the risk assessment, psychiatric nurses documented whether patients had behaved aggressively towards others or whether they had deliberately damaged property. They also noted the target of aggression, whether towards staff, patients or property. Results: The predictive validity of the DASA varied according to the type and target of aggression. The prediction of any aggressive behaviour, irrespective of type of aggression or target, was significantly greater than chance (AUC = 0.69). The strongest predictive accuracy (AUC = 0.80) was for physical aggression towards staff. Conclusions: These results suggest that imminent aggression in psychiatric hospitals may be able to be accurately predicted by psychiatric nurses using a structured risk assessment instrument. PMID- 20001375 TI - Patient aggression perceived by community support workers. AB - Objective: Aggression by patients is a known risk factor for hospital workers. Within New Zealand, the bulk of ongoing care for physical and mental disabilities and health issues is not hospital based, but contracted to various non governmental agencies. The rate of client aggression towards care workers from these organizations, to our knowledge, has not been assessed. Method: Two hundred and forty-two support workers in non-governmental agencies caring for people with disabilities responded to an anonymous mailed survey on client aggression, personal distress, and communication style. Results: Most support workers did experience verbal forms of aggression or destructive behaviour, fewer experienced physical aggression, and a minority were injured, sexually harassed, stalked or harassed by means of formal complaint. The median total violence score was five (interquartile range 12.25). A higher total violence score (using the POPAS-NZ) was associated with age and gender, the primary disability of clients, and the numbers of hours worked. The length of time worked was not associated with total violence risk. Communication style, after correcting for other factors, was a predictor of aggression. Almost 6% of care workers reported distress symptoms at a level associated with clinically significant stress reactions. Conclusions: Patient aggression is common among care workers, and can cause distress in the minority. We suggest that further research to clarify risk factors and develop interventions for care workers is needed. PMID- 20001376 TI - Academics of the future? A survey of final year medical students. AB - Objective: The aim of this study was to survey final year medical students at an Australian medical school about their views and experiences regarding academic medicine Methods: A four page questionnaire was developed and administered to each group of final (6th) year medical students upon completion of their psychiatry rotation Results: For 82% clinical teaching would probably or definitely be part of their future careers, while 63% expected that they would undertake formal teaching. Thirty-three percent of students anticipated limited or no involvement in research. While 26% of students expected that they would undertake a higher degree, only 13% were interested in pursuing an academic career Conclusions: The dearth of academics in medicine and in psychiatry in particular is of considerable concern. This study provides some possible avenues for further exploration to address this professional challenge. PMID- 20001377 TI - Expanding psychiatry training: Australian specialists' and trainees' perceived gaps in experiences and settings. AB - Objective: The Royal Australian and New Zealand College of Psychiatrists (RANZCP) is working together with the Australian Federal Government to expand the range of training experiences and settings with the object of producing better equipped and better distributed psychiatrists to address the rising prevalence of mental disorder. To assist in this process, the College sought the views of its Fellows (psychiatrists) and trainees concerning important gaps in clinical experience and the settings best suited to filling them. Method: An electronic survey was sent to all Australian RANZCP psychiatrists and trainees in June 2008. Results: Only 15% of psychiatrists and 28% of trainees responded to the survey. There was striking concordance, however, that psychological therapies required much greater emphasis and that this experience would best be acquired in the private healthcare system. Conclusions: These findings support the recommendations of the Medical Specialist Training Steering Committee's report on expanded settings, suggesting that RANZCP members are likely to engage enthusiastically in the expanded specialist training program. It will be important to ensure that trainees are well supervised in their treatment of anxiety, affective and substance abuse disorders. PMID- 20001378 TI - I just want you to be happy: preventing and tackling teenage depression. PMID- 20001379 TI - Suicide Prevention. PMID- 20001380 TI - College Congress and pharma. PMID- 20001381 TI - Clinical supervision: which components are most valued by trainees? PMID- 20001383 TI - The tremulous gun hand. PMID- 20001382 TI - Quetiapine causing peripheral oedema. PMID- 20001384 TI - Recruitment, retention and the Psychiatrist Training Initiative. PMID- 20001386 TI - From the Honorary Secretary. PMID- 20001385 TI - From the President. PMID- 20001388 TI - ? PMID- 20001387 TI - From the Chief Executive Officer. PMID- 20001389 TI - Our reviewers. PMID- 20001390 TI - Cardiovascular risk factors in hypertensive patients born in Northern Africa and living in France. AB - Northern Africans constitute one of the largest immigrant groups in France; however, limited data are available on their cardiovascular risk factors. We carried out a retrospective cross-sectional study in a French hypertension unit, comparing 719 patients born in Northern Africa to 3558 controls born in Europe, individually matched for age and sex. Using a Bonferroni adjusted alpha level=0.001, we found no significant difference between the groups for blood pressure levels, anti-hypertensive treatment, prevalence of target organ damage or the proportion of patients with secondary hypertension. However, patients of both sexes born in Northern Africa were less likely to take regular physical exercise than their controls. In addition, women born in Northern Africa were less often current or former smokers than their European counterparts (19.9% vs 30.5%, p < 0.001), but had a higher body mass index (28.5 vs 26.8 kg/m(2), p < 0.001) and a higher prevalence of diabetes (19.1% vs 8.9%, p < 0.001 after adjusting for BMI). These results suggest that targeted lifestyle interventions, including regular physical exercise, could be proposed to prevent weight gain and decrease the incidence of diabetes in hypertensive women born in Northern Africa and living in western countries. PMID- 20001391 TI - Aortic stiffness in normal and hypertensive pregnancy. AB - The objective of this study was to examine whether aortic stiffness, as assessed by pulse wave analysis, could reliably discriminate between normal and hypertensive pregnancies. One hundred pregnant women were studied: five with severe pre-eclampsia, 27 with gestational hypertension, 14 with chronic hypertension and 54 with normal pregnancy. Central hemodynamic parameters were obtained by an applanation tonometry and included central aortic systolic blood pressure (CSBP), central aortic diastolic blood pressure (CDBP), augmentation pressure (AP), augmentation index (AIx), AIx corrected to a heart rate of 75 (AIx@75) and time to reflection (Tr). All measures of aortic stiffness, including AP, AIx and AIx@75 were significantly higher in women with gestational hypertension and pre-eclampsia compared with normal pregnancies and women with chronic hypertension (p < 0.05 for all comparisons). There were no significant differences between normal pregnancies and women with chronic hypertension (p > 0.05 for all comparisons). Tr was significantly shorter in women with pre eclampsia and gestational hypertension compared with normal pregnancies (p < 0.05). Aortic stiffness, as assessed by pulse wave analysis, is significantly increased in women with pre-eclampsia and gestational hypertension but not in treated women with chronic hypertension. Pulse wave analysis has a potential as a screening tool in women at high risk for pre-eclampsia. The final role of this method should be determined in prospective studies. PMID- 20001392 TI - Effect of carvedilol on coronary flow reserve in patients with hypertensive left ventricular hypertrophy. AB - OBJECTIVE: Patients with hypertensive left-ventricular hypertrophy (LVH) have lower coronary flow reserve (CFR). Whether carvedilol can improve CFR of patients with hypertensive LVH is unknown. We aimed to investigate the effects of carvedilol on CFR in patients with hypertensive LVH. METHODS: Sixty-three patients were randomly divided into two groups for treatment with carvedilol or metoprolol. The peak diastolic coronary flow velocity in the left anterior descending coronary artery at rest and at maximal vasodilation with dipyridamole infusion was recorded by transesophageal echocardiography (TEE), then CFR was calculated at baseline and at the end of 6 months of therapy. Left-ventricular mass index (LVMI) was calculated by 2-D echocardiography. Endothelium-dependent and -independent reactivity of the brachial artery was measured. Levels of plasma endothelin-1 (ET1), nitric oxide (NO) and other metabolites were monitored and analyzed before and after 6-month therapy. RESULTS: Both blood pressure and heart rate decreased significantly in the two treatment groups after therapy (p<0.05). With carvedilol treatment, LVMI was lower (p<0.05), endothelium function of the brachial artery was higher (p<0.05), and peak diastolic coronary flow velocity at rest and at maximal vasodilation after dipyridamole infusion was significantly higher (p<0.05) than with metoprolol treatment, which led to a significantly higher CFR (p<0.05). Changes in CFR and LVMI with carvedilol treatment were inversely correlated (R(2)=0.474, p=0.036). With carvedilol treatment, plasma level of ET-1 was lower, but that of NO was significantly higher than with metoprolol treatment (both p<0.05). CONCLUSIONS: The CFR of patients with hypertensive LVH but not coronary artery disease could increase with 6-month carvedilol therapy. PMID- 20001394 TI - Evolution and psychiatry. PMID- 20001393 TI - Chronic obstructive pulmonary disease and cardiovascular mortality in elderly subjects from general population. AB - AIMS: To ascertain whether chronic obstructive pulmonary disease (COPD) is an independent risk factor for cardiovascular (CV) mortality in the elderly subjects from general population. METHODS: 3282 subjects of the Northern Italy aged > or =65 years were followed up for 12 years in the frame of the CArdiovascular STtudy in the ELderly. Multivariate stepwise proportional hazard Cox regression was therefore used to identify the prognostic role of COPD on CV mortality in hypertensive (HT) and normotensive (NT) subjects. The hazard ratio (HR) of COPD with 95% confidence interval (CI) for mortality was adjusted for confounders in both genders. RESULTS: COPD resulted to be an independent predictor of CV mortality (HR 1.34, CI 1.13-1.61) in HT but not in NT subjects. This was evident both in men (HR 1.44, 1.25-1.95) and women (HR 1.32, CI 1.14-1.53); pulse pressure (PP) was directly related and anti-hypertensive therapy inversely related to risk of CV mortality, an association that was greater in subjects with than without COPD. CONCLUSION: COPD should be included in the computation of global risk in HT subjects. PP is the main BP component in increasing CV risk in subjects with COPD. Controlled trials should be performed to evaluate the pressor targets to be reached in HT subjects with COPD, with the aim of decreasing their CV risk. PMID- 20001395 TI - Evolution of human brain functions: the functional structure of human consciousness. AB - The functional structure of self-aware consciousness in human beings is described based on the evolution of human brain functions. Prior work on heritable temperament and character traits is extended to account for the quantum-like and holographic properties (i.e. parts elicit wholes) of self-aware consciousness. Cladistic analysis is used to identify the succession of ancestors leading to human beings. The functional capacities that emerge along this lineage of ancestors are described. The ecological context in which each cladogenesis occurred is described to illustrate the shifting balance of evolution as a complex adaptive system. Comparative neuroanatomy is reviewed to identify the brain structures and networks that emerged coincident with the emergent brain functions. Individual differences in human temperament traits were well developed in the common ancestor shared by reptiles and humans. Neocortical development in mammals proceeded in five major transitions: from early reptiles to early mammals, early primates, simians, early Homo, and modern Homo sapiens. These transitions provide the foundation for human self-awareness related to sexuality, materiality, emotionality, intellectuality, and spirituality, respectively. The functional structure of human self-aware consciousness is concerned with the regulation of five planes of being: sexuality, materiality, emotionality, intellectuality, and spirituality. Each plane elaborates neocortical functions organized around one of the five special senses. The interactions among these five planes gives rise to a 5 x 5 matrix of subplanes, which are functions that coarsely describe the focus of neocortical regulation. Each of these 25 neocortical functions regulates each of five basic motives or drives that can be measured as temperaments or basic emotions related to fear, anger, disgust, surprise, and happiness/sadness. The resulting 5 x 5 x 5 matrix of human characteristics provides a general and testable model of the functional structure of human consciousness that includes personality, physicality, emotionality, cognition, and spirituality in a unified developmental framework. PMID- 20001396 TI - Ethological approaches to psychiatric disorders: focus on depression and schizophrenia. AB - Behavioural observation of psychiatric patient groups using ethological methodology has never been a mainstream approach in psychiatry. In the present review article it is argued that the assessment of non-verbal behaviour in psychiatric disorders has much to offer to clinicians. Based on a Medline survey, the literature on ethological observation of patients with depression and schizophrenia was reviewed. Ethological observation of psychiatric patient groups has the potential to improve prediction of course and outcome of severe psychiatric disorders at a very early stage. Disadvantages of the ethological approach lie within its technical requirements and its inherent time-consuming evaluation of data. A full appreciation of psychiatric disorders may require answers to questions to the proximate causes and evolutionary (Darwinian) trajectories of behaviour, an approach that may well be expanded to maladaptive cognition and emotion. PMID- 20001397 TI - Evolution of the human CNS cholineric system: has this resulted in the emergence of psychiatric disease? AB - The aim of the present study was to review the available literature on the evolutionary processes that have led to the development of the human central nervous system (CNS) cholinergic system and to test the hypothesis that such processes may have contributed to the emergence of psychiatric diseases. First, it is clear that the molecular components that have come together to form the cholinergic system in the human CNS initially had functions that were not involved in neurotransmission. Indeed, components of the cholinergic system of the human CNS may be present in nearly all forms of life and may have existed since life began. Significantly, components of the human CNS cholinergic system had begun to interact with each other millions of years ago, long before the appearance of any nervous system. Moreover, there are data to suggest that changes in the mammalian CNS cholinergic system can involve involved gene and environment interactions. Evidence is available to support the notion that the changes in functioning of the human CNS cholinergic system in individuals with psychiatric disorders may have resulted from both Darwinian and Lamarckian concepts of evolution. PMID- 20001398 TI - Darwinian dynamics of depression. AB - Darwin's theory of sexual selection offers a challenge to psychology and psychiatry. We select each other, and have been doing so since social life first evolved. But who is selected and what happens to those who are not selected? What social structures have evolved to contain the unselected? What behaviours have evolved to manage the selection process? How do the selected relate to the unselected and what behaviours have evolved to manage this asymmetry in social relations? What mental states have evolved to characterize the selected and the unselected? These questions should be kept in mind when we observe and study the social structures, behaviours and mental states that we see displayed before us in all the variety of nature. It is suggested that a significant amount of current psychiatric disorder, especially depressive states and both social and generalized anxiety disorder, have evolved because they managed the processes of being unselected and de-selected, and maintained the unselected in that social role without loss of life or physical incapacity, and enabled the unselected to contribute to general social well-being. PMID- 20001399 TI - Post-traumatic stress disorder: evolutionary perspectives. AB - Fear is the key emotion of post-traumatic stress disorder (PTSD). Fear's evolved function is motivating survival via defensive behaviours. Defensive behaviours have been highly conserved throughout mammalian species; hence much may be learned from ethology. Predation pressure drove the early evolution of defences, laying foundations in the more ancient brain structures. Conspecific (same species) pressure has been a more recent evolutionary influence, but along with environmental threats it has dominated PTSD research. Anti-predator responses involve both avoiding a predator's sensory field and avoiding detection if within it, as well as escape behaviours. More effective avoidance results in less need for escape behaviours, suggesting that avoidance is biologically distinct from flight. Recognizing the predation, environmental and conspecific origins of defence may result in clearer definition of PTSD phenomena. Defence can also be viewed in the stages of no threat, potential threat, encounter and circa strike. Specific defences are used sequentially and according to contexts, loosely in the order: avoidance, attentive immobility, withdrawal, aggressive defence, appeasement and tonic immobility. The DSM-IV criteria and PTSD research show substantial congruence with the model proposed: that PTSD is a disorder of heightened defence involving six key defences used in conjunction with vigilance and risk assessment according to contexts. Human research is reviewed in this respect with reference to laboratory and wild animal observations providing new insights. Understanding individual perceptual issues (e.g. predictability and controllability) relevant to these phenomena, combined with defence strategy recalibration and neuronal plasticity research goes some way to explaining why some traumatized individuals develop PTSD when others do not. PMID- 20001400 TI - Drug use and addiction: evolutionary perspective. AB - Drug use creates a significant amount of harm in modern societies. From an evolutionary perspective, the pervasive use of drugs and the ongoing risk of drug addiction can be explained in terms of the action of drugs on evolved motivational-emotional systems. Addiction arises through interaction of these evolutionarily ancient systems, designed to promote the pursuit of natural rewards, and contemporary environments where purified and potent forms of drugs are readily available. This evolutionary analysis is extended to account for developmental patterns in problem drug use, and to explain the existence of behavioural addictions, such as problem gambling. The paper concludes by considering some of the clinical and public policy implications of the evolutionary perspective presented. PMID- 20001401 TI - Abnormalities in MRI signal intensity in schizophrenia associated with idiopathic unconjugated hyperbilirubinemia. AB - OBJECTIVE: The purpose of the present study was to examine whether patients with idiopathic unconjugated hyperbilirubinemia (Gilbert's syndrome; GS) have specific changes in signal intensity on magnetic resonance imaging (MRI). METHODS: Axial 5 mm-thick T1-weighted and T2-weighted MRI was acquired from schizophrenia patients with GS (n = 24) and schizophrenia patients without GS (n = 60). All patients were diagnosed according to DSM-IV criteria and were compared with age- and sex matched healthy controls without GS (n=60) and controls with GS (n=36). Signal intensity in the hippocampus, amygdala, caudate, putamen, globus pallidus, thalamus, anterior cingulate gyrus, posterior cingulate gyrus, insular cortex, and cerebellum was measured in relation to the signal intensities of the vitreous body. RESULTS: Compared to both schizophrenia patients without GS and the control subjects without or with GS, the schizophrenia patients with GS had significantly decreased signal intensity in almost all the regions measured on T1-weighted MRI. On T2-weighted MRI, the schizophrenia patients with GS had significantly increased signal intensity in almost all the regions measured compared to both schizophrenia patients without GS and the control subjects without or with GS. CONCLUSION: Patients with schizophrenia-associated GS have specific changes in signal intensity on T1- and T2-weighted MRI, suggesting that schizophrenia with GS produces changes specifically in the frontotemporal cortex, limbic system, and basal ganglia. PMID- 20001402 TI - Community attitudes towards mental health interventions for healthy people on the basis of genetic susceptibility. AB - OBJECTIVES: The aim of the present study was to evaluate, using serotonin transporter genotyping as an example, the preparedness of individuals from an urban general population identified with hypothetical genetic risk for a depressive disorder to moderate risk through cognitive or behavioural intervention. It also evaluated endorsement of genetic and environmental causal attributions of mental illness. METHOD: A qualitative approach using focus group methodology was selected as most appropriate because these issues are relatively unexplored. Participants (n=36) aged > or =18 years from metropolitan Sydney discussed their understanding of the role of genetic and environmental risk factors in mental illness and attitudes towards pre-symptomatic interventions based on genetic risk information. RESULTS: Thirty-six participants attended four focus groups involving 8-10 participants per group. Participants predominantly viewed genetic risk factors for depression as predisposing rather than causal, with environmental risk factors acting as triggers. Hypothetical identification with a genetic variant suggesting predisposition to depression prompted strong interest in seeking further information about predictive genetic testing from medical professionals, willingness to reduce life stress, drugs and alcohol intake, willingness to increase exercise, and willingness to undertake cognitive and behavioural interventions at a pre-symptomatic stage. Mixed views prevailed as to whether stress was a modifiable risk factor. Preventive intervention at a presymptomatic stage of depression was viewed negatively in a minority of participants due to a fatalistic attitude towards a genetic predisposition and attitudes that intervention was futile in the absence of symptoms. CONCLUSIONS: There is a likely public demand for preventive mental health interventions for healthy people on the basis of genetic susceptibility if predictive genetic testing becomes available in psychiatry. The findings have implications for general practitioner and public education about predictive genetic testing for susceptibility to common multifactorial disorders for at-risk groups. PMID- 20001403 TI - Effectiveness of community treatment orders for treatment of schizophrenia with oral or depot antipsychotic medication: changes in problem behaviours and social functioning. AB - OBJECTIVE: Involuntary outpatient commitment (IOC) has been in use in various countries for a number of years and has recently been implemented (in the form of supervised community treatment) in England and Wales. Several studies indicate that IOC reduces relapse and readmission rates and decreases length of stay on inpatient units in patients diagnosed with schizophrenia. The aim of the present study was to examine whether the use of IOC in the Australian context, in the form of community treatment orders (CTOs), may be associated with a reduction in problem behaviours and improved social functioning. METHOD: A naturalistic retrospective mirror image study of case notes, with each case serving as its own control, was used. Behavioural and social outcomes were examined: episodes of aggression and suicidal and self-harming behaviour, episodes of homelessness, frequency of contact with family members and overall quality of relationship between family and patient, and employment status. RESULTS: Ninety-four sets of case notes were identified as meeting the criteria for inclusion. The number of episodes of aggression was found to be halved from the year before the CTO to the subsequent year (p<0.0001). Significant reductions in the number of episodes of homelessness were experienced by patients (p<0.05) when the pre-CTO year was compared with the CTO year. CONCLUSION: A CTO may contribute to improved outcomes related to patient quality of life. This may be seen to mitigate concerns about infringement of civil rights. PMID- 20001404 TI - Self-enucleation of one eye. PMID- 20001406 TI - Evidence on the use of platelet-rich plasma for diabetic ulcer: a systematic review. AB - The aim of topical wound treatment is to favor efficient, rapid, and safe healing. The platelet-rich plasma (PRP) has been used for wound treatment since it contains various platelet growth factors. The objective of the present study was to collect evidence regarding the use of PRP for the topical treatment of chronic leg ulcers. For this purpose, a systematic review of the literature was performed according to the steps recommended by the Cochrane Collaboration with studies published until July 2008. Among 18 selected studies, 7 (39%) of these studies were randomized clinical. Five of the seven randomized clinical trials studied ulcers of diabetic etiology. The results of meta-analysis showed that PRP favors the healing process (95% CI: 2.94-20.31). In conclusion, the present systematic review and meta-analysis show that there is a scientific evidence regarding favorable outcomes of the use of PRP for the treatment of diabetic ulcer. PMID- 20001407 TI - Ion men and their mettle. PMID- 20001408 TI - Is lithium in a class of its own? A brief profile of its clinical use. AB - Lithium is a unique and effective psychotropic agent with a long-standing history of clinical use yet it is increasingly overlooked in lieu of newer agents. The purpose of the present paper was to succinctly review the therapeutic profile of lithium particularly with respect to the treatment of mood disorders and consider its unique properties and clinical utility. A comprehensive literature review pertaining to lithium was undertaken using electronic database search engines to identify relevant clinical trials, meta-analyses and Cochrane reviews. In addition articles and book chapters known to the authors were carefully reviewed, and the authors appraised published guidelines. The evidence from these sources was rated using National Health and Medical Research Council evidence levels and synthesized according to phenotype and mood states. In addition, the authors have drawn upon published guidelines and their own clinical experience. Lithium has specificity for mood disorders with proven efficacy in the treatment of both unipolar depression and bipolar disorder. The recommendations are based predominantly on Level I evidence, but its clinical use has to be tempered against potential side-effects and the need for ongoing monitoring. In practice, lithium should be considered a first-line option in bipolar disorder, especially in prophylaxis and when onset of action is not an imperative. Lithium has been in use in modern medicine for 60 years and as such has been tried and tested across the full range of mood disorders. Arguably, lithium is the only true mood stabilizer and because of its unique properties is in a class of its own. PMID- 20001409 TI - Longitudinal assessment of neuropsychological function in major depression. AB - Neuropsychological impairment is a core component of major depression, yet its relationship to clinical state is unclear. The aims of the present review were to determine which neuropsychological domains and tasks were most sensitive to improvement in clinical state in major depression and to highlight the methodological issues in such research. Studies that included a baseline and at least one follow-up neuropsychological testing session in adults with major depression were identified using MEDLINE, Web of Science and ScienceDirect databases. Thirty studies were included in the review. Findings in younger adult populations suggested that improvement in mood was most strongly related to improved verbal memory and verbal fluency, while measures of executive functioning and attention tended to remain impaired across treatment. In late life major depression, improved psychomotor speed was most closely related to treatment response, but there was much inconsistency between study findings, which may be due to methodological issues. In major depression, particular neuropsychological domains are more strongly related to clinical state than others. The findings from the present review suggest that the domains most sensitive to clinical state are verbal learning and memory, verbal fluency and psychomotor speed. In contrast, measures of attention and executive functioning perhaps represent more trait-like markers of major depression. With further methodologically sound research, the changes in neuropsychological function associated with treatment response may provide a means of evaluating different treatment strategies in major depression. PMID- 20001410 TI - Use of administrative data for the surveillance of mood and anxiety disorders. AB - OBJECTIVE: There is increasing interest in the use of administrative data for surveillance and research in Australia. The purpose of the present study was to evaluate the usefulness of such data for the surveillance of mood and anxiety disorder using databases from the following Canadian provinces: British Columbia, Ontario, Quebec and Nova Scotia. METHOD: A population-based record-linkage analysis was done using data from physician billings and hospital discharge abstracts, and community-based clinics using a case definition of ICD-9 diagnoses of 296.0-296.9, 311.0, and 300.0-300.9. RESULTS: The prevalence of treated mood and/or anxiety disorder was similar in Nova Scotia, British Columbia, and Ontario at approximately 10%. The prevalence for Quebec was slightly lower at 8%. Findings from the provinces showed consistency across age and sex despite variations in data coding. Women tended to show a higher prevalence overall of mood and anxiety disorder than men. There was considerably more variation, however, when treated anxiety (300.0-300.9) and mood disorders (296.0-296.9, 311.0) were considered separately. Prevalence increased steadily to middle age, declining in the 50s and 60s, and then increased after 70 years of age. CONCLUSIONS: Administrative data can provide a useful, reliable and economical source of information for the surveillance of treated mood and/or anxiety disorder. Due to the lack of specificity, however, in the diagnoses and data capture, it may be difficult to conduct surveillance of mood and anxiety disorders as separate entities. These findings may have implications for the surveillance of mood and anxiety disorders in Australia with the development of a national network for the extraction, linkage and analysis of administrative data. PMID- 20001411 TI - Suicide in the Northern Territory, 2001-2006. AB - OBJECTIVE: The aim of the present study was to characterize suicide in the Northern Territory (NT) for the 6 year period 1 January 2001-31 December 2006. METHOD: Suicide death rates by area (Australia, NT), sex, and Indigenous status for the period of 2001-2006 were obtained from the National Coroners Information System through the Victorian Institute of Forensic Medicine. Population figures were obtained from the Australian Bureau of Statistics. RESULTS: For this period the suicide rate of NT (21.6 per 100 000) was double the national rate ( 11 ), and the rate for NT Indigenous people (36.7) was significantly higher (p < 0.001) than that of NT non-Indigenous people (14.7). There was a decline in the suicide rate for Australia of -8.6%; for NT, -6.3%; for NT non-Indigenous, -3.3%; and for NT Indigenous, -9.0%. The sex difference was significant (p < 0.001) for both Australia and the NT with male subjects having a higher suicide rate than female subjects. With respect to age group, some evidence suggested an earlier peak for NT Indigenous compared to NT non-Indigenous people. Hanging was more common in NT than in the rest of the country and accounted for 87% of Indigenous suicide. CONCLUSION: Suicide is more common in NT than in Australia generally, and more common in NT Indigenous than NT non-Indigenous people. There is evidence of a decline in suicide rates across the board. PMID- 20001412 TI - Antidepressant dispensing trends in New Zealand between 2004 and 2007. AB - OBJECTIVE: The aim of the present study was to explore antidepressant dispensing trends in New Zealand over a 3 year period (2004-2007) with a focus on trends by age, gender, ethnicity, District Health Board and type of antidepressant. METHOD: Prevalence of antidepressant agents dispensed in the years ending June 2004-2005, 2005-2006 and 2006-2007 was calculated separately for the population aged 15-24, 24-44, 45-64 and 65-100 years, (> =65) by gender. The f rst occurrence of antidepressant dispensing for each individual in each year was extracted to calculate age-specific antidepressant dispensing rates standardized to the 2006 Census population for the New Zealand European/Other (NZEO) ethnic group. RESULTS: The 12 month prevalence of antidepressant dispensing among the total population aged 15-100 years increased from 7.36% in 2004-2005 to 8.21% in 2005 2006 and to 9.39% in 2006-2007 Selective serotonin re-uptake inhibitor (SSRI) and tricyclic antidepressants (TCA) accounted for >90% of all antidepressants dispensed each year. SSRIs were more frequently prescribed to young adults (15-24 years) than TCAs, although this relation changed with increasing age. In women aged > =65, TCAs were more commonly dispensed than SSRIs, while among men in the same age group, dispensing patterns were similar for both agents. Age-specific dispensing rates were higher among female than male subjects, and the NZEO combined populations were dispensed significantly more antidepressant agents than Maori or Pacific peoples. Considerable regional variations in antidepressant dispensing were found by ethnicity and gender. CONCLUSION: There are considerable variations in antidepressant dispensing in NZ, and the rate of dispensing has increased over the 3 years of the present study. Further work is required to investigate the different rates of antidepressant dispensing found between ethnic groups. Depression is strongly associated with suicide and the trend of increased dispensing of antidepressants found in the present study should continue to have a positive effect on suicide rates in NZ. PMID- 20001413 TI - Factor structure of manic symptoms. AB - OBJECTIVE: The purpose of the present study was to identify the factor structure of manic symptoms as assessed by the Scale for Manic States (SMS). METHOD: The pattern of symptoms in a group of 225 adults with an ICD-10-DCR diagnosis of manic episode was studied. A factor analysis was conducted of the broad range of psychiatric symptoms covered by SMS. A principal component analysis followed by oblimin rotation was performed. RESULTS: Six eigenvalues were greater than unity and parallel analysis indicated four factors. After observing the scree plot a five-factor solution seemed appropriate. Nevertheless, a six-factor solution was chosen that described the data appropriately and was clinically relevant. The six factors (psychosis, irritability aggression, dysphoria, accelerated thought stream, hedonia, and hyperactivity) captured 59.29% of the total variance. CONCLUSION: A six-factor solution explains the clinical dimensions of mania in the present sample and dysphoria appeared as a separate factor. PMID- 20001414 TI - Prevalence and correlates of depression with atypical symptoms in Hong Kong. AB - OBJECTIVE: Previous studies of atypical depression have been conducted in Western communities. There has been no community-based epidemiological study of atypical depression that covers the entire spectrum of bipolar disorders. The aim of the present study was to examine the 12 month prevalence of atypical depression and the differences in demographic and clinical profiles between depressed patients with and without atypical symptoms in the Chinese adult population of Hong Kong. METHOD: A random sample of 3016 Chinese adults completed a telephone-based structured interview that examined their 12 month prevalence of major depressive episode. Atypical depression was defined as major depressive episode with hypersomnia and increased weight or appetite. Major depressive episode with and without atypical symptoms were compared on sociodemographic variables, number of depressive and manic/hypomanic symptoms, proportion of having lifetime mania/hypomania, suicidality, family psychiatric history, help-seeking behaviour, level of distress and role impairment. RESULTS: The 12 month prevalence of atypical depression was 1.3%. Compared to non-atypical depression, atypical depression was associated with female gender, soft (subthreshold) bipolar II disorder, family psychiatric history, higher suicidality, more help-seeking from psychiatrists, and more depressive and manic/hypomanic symptoms. There was no difference in levels of distress or impairment. CONCLUSIONS: Although limited by the lack of detailed information on comorbidity, bipolar family history, and age of onset, the findings support the hypothesis that atypical depression among Chinese people in Hong Kong exhibited prevalence and correlates similar to those found in Western epidemiological and clinical studies. Further research is warranted to examine its association with hypomania and how atypical depression may occupy a nosological position between typical unipolar depression and bipolar spectrum disorders. PMID- 20001415 TI - Subjective effects of cannabis before the first psychotic episode. AB - OBJECTIVE: The aim of the present study was to gain more insight into the positive and negative effects of cannabis in the prodromal phase of schizophrenia and in the ultra-high-risk (UHR) state for psychosis. METHOD: A theory-driven questionnaire was used to examine subjective effects in the prodromal phase in male subjects with a recent onset of schizophrenia or related disorder (n = 52) and in the UHR state in help-seeking male subjects screened for being at UHR for psychosis (n = 17); both groups were compared to cannabis-using controls from the general population (n=52). RESULTS: Recent-onset patients and UHR subjects reported feeling more anxious, depressed and suspicious immediately after cannabis use. Some patients also reported feeling less depressed after cannabis use. Recent-onset patients reported increased visual and acoustic hallucinations, and confusion after cannabis use. Of the recent-onset patients 37% reported that their very first psychotic symptoms occurred during cannabis intoxication. Long term effects of cannabis reported more often by both patient groups were depression, less control over thoughts and social problems. CONCLUSIONS: These results suggest that schizophrenia patients in the prodromal phase and subjects at UHR for psychosis are more sensitive to some negative effects of cannabis, in particular psychotic effects, compared to cannabis users from the general population. Although limited by the retrospective design in the recent-onset patients, the present study adds qualitative evidence to longitudinal studies that suggest that cannabis is a component cause in the onset of the first psychotic episode. Further studies are needed on the objective and subjective effects of cannabis in UHR subjects. PMID- 20001416 TI - Does school bullying affect adult health? Population survey of health-related quality of life and past victimization. AB - OBJECTIVE: The purpose of the present study was to determine the proportion of adult South Australians who report having experienced school bullying and to examine the relationship between past victimization and adult health-related quality of life. METHOD: A representative sample (n=2833) of metropolitan and country South Australian adults were asked in a face-to-face interview whether they had experienced bullying when they were at school. Health-related quality of life was measured using the Medical Outcomes Study Short Form 36-item health survey questions (SF-36). Regression analyses (linear and logistic) were performed, taking into account survey weights. RESULTS: Nearly one-fifth of adults reported having experienced bullying when they were at school. Older persons and those born overseas were less likely to report having been bullied. Those reporting that they had been bullied experienced significantly poorer mental and physical health compared to those who had not been bullied. CONCLUSIONS: Adults commonly reported experiencing bullying while at school and these reports were associated with lower health-related quality of life in adulthood. School bullying needs further investigation as a preventable cause of mental health problems across the lifespan. PMID- 20001417 TI - BraveHeart begins: pilot results of group cognitive behaviour therapy for depression and anxiety in cardiac patients. AB - OBJECTIVE: In a general medical hospital with limited mental health resources, a clinical need arose for an effective treatment for depression and anxiety symptoms in cardiac rehabilitation patients. METHOD: A total of 628 cardiac rehabilitation outpatients at a tertiary care centre were screened with the Hospital Anxiety and Depression Scale at week 4 of their programme, and 182 of 558 responders (33%) scored > or =8 on the Depression and/or Anxiety subscales. A 6 week group cognitive behaviour therapy programme was developed to assist these identified patients. The resulting programme, BraveHeart, was piloted on 39 patients still experiencing significant symptoms of depression or anxiety at the end of rehabilitation. Patients were assessed at baseline, after treatment, 1 and 6 months after treatment using the Hospital Anxiety and Depression Scale and the Beck Depression Inventory-II. RESULTS: Significant improvement in levels of depression and anxiety symptoms occurred from baseline to post-treatment assessment, and this change was maintained at 6 months, with moderate-strong effect sizes. Feedback from the group members was positive. CONCLUSIONS: A specialized group treatment programme for cardiac patients with existing depression and anxiety was created that resulted in significant reductions in symptoms. PMID- 20001418 TI - Nutrition knowledge in young women with eating disorders in Australia and Singapore: a pilot study. AB - OBJECTIVE: The aim of the present study was to compare nutrition knowledge levels in young women with and without an eating disorder (ED) in two countries. METHOD: Women with a clinical ED (n = 55) and healthy control women (n = 99) in Australia and Singapore completed a Nutritional Knowledge Questionnaire, acculturation questionnaire and demographics survey. Nutrition knowledge was analysed in terms of clinical status, cultural group, acculturation, socioeconomic status and education level. results: Women with EDs had greater knowledge than controls, but the magnitude of the difference was small. Greater acculturation to Western culture was associated with greater knowledge. CONCLUSIONS: The difference in nutrition knowledge between women with and without EDs is unlikely to be of clinical importance. The findings may reflect today's ubiquitous availability of nutrition information. PMID- 20001419 TI - Utility of telepsychiatry for Aboriginal Australians. PMID- 20001420 TI - De novo induction of obsessive-compulsive symptoms with quetiapine in a patient with borderline personality disorder. PMID- 20001421 TI - Systems biology of lupus: mapping the impact of genomic and environmental factors on gene expression signatures, cellular signaling, metabolic pathways, hormonal and cytokine imbalance, and selecting targets for treatment. AB - Systemic lupus erythematosus (SLE) is characterized by the dysfunction of T cells, B cells, and dendritic cells, the release of pro-inflammatory nuclear materials from necrotic cells, and the formation of antinuclear antibodies (ANA) and immune complexes of ANA with DNA, RNA, and nuclear proteins. Activation of the mammalian target of rapamycin (mTOR) has recently emerged as a key factor in abnormal activation of T and B cells in SLE. In T cells, increased production of nitric oxide and mitochondrial hyperpolarization (MHP) were identified as metabolic checkpoints upstream of mTOR activation. mTOR controls the expression T cell receptor-associated signaling proteins CD4 and CD3zeta through increased expression of the endosome recycling regulator Rab5 and HRES-1/Rab4 genes, enhances Ca2+ fluxing and skews the expression of tyrosine kinases both in T and B cells, and blocks the expression of Foxp3 and the generation of regulatory T cells. MHP, increased activity of mTOR, Rab GTPases, and Syk kinases, and enhanced Ca2+ flux have emerged as common T and B cell biomarkers and targets for treatment in SLE. PMID- 20001422 TI - The biology of reactive intermediates in systemic lupus erythematosus. AB - Systemic lupus erythematosus (SLE) is an autoimmune syndrome marked by autoantibody production. Innate immunity is essential to transform humoral autoimmunity into the clinical lupus phenotype. Nitric oxide (NO) is a membrane- permeable signaling molecule involved in a broad array of biologic processes through its ability to modify proteins, lipids, and DNA and alter their function and immunogenicity. The literature regarding mechanisms through which NO regulates inflammation and cell survival is filled with contradictory findings. However, the effects of NO on cellular processes depend on its concentration and its interaction with reactive oxygen. Understanding this interaction will be essential to determine mechanisms through which reactive intermediates induce cellular autoimmunity and contribute to a sustained innate immune response and organ damage in SLE. PMID- 20001424 TI - Commentary on pharmacotherapy of regional melanoma therapy. AB - IMPORTANCE TO THE FIELD: Regional therapy continues to be the workhorse for the treatment of regional metastases and unresectable recurrences of melanoma limited to a limb. These approaches also offer an excellent opportunity for the study of disease biology and new drug delivery, pharmacokinetics and pharamacotherapeutics. AREAS COVERED IN THIS EDITORIAL: Utility of regional therapy as an area of study, benefits of both isolated limb infusion (ILI) and hyperthermic isolated limb perfusion (HILP) are discussed. The limitations of both approaches to regional therapy are also referenced. WHAT THE READER WILL GAIN: This editorial serves as a companion to the peer-reviewed paper which comprehensively reviews the subject of regional therapy by Tyler et al. It offers a brief commentary on the utility of regional therapies (ILI and HILP) for extremity in-transit melanoma and their role in investigating new therapeutic modalities. TAKE HOME MESSAGE: Regional therapy is an excellent therapeutic modality for disease limited to a limb and furthermore serves as an excellent model for scientific investigation, both clinical and translational. PMID- 20001423 TI - Immune pathology associated with altered actin cytoskeleton regulation. AB - The actin cytoskeleton plays a crucial role in a variety of important cellular processes required for normal immune function, including locomotion, intercellular interactions, endocytosis, cytokinesis, signal transduction, and maintenance of cell morphology. Recent studies have uncovered not only many of the components and mechanisms that regulate the cortical actin cytoskeleton but have also revealed significant immunopathological consequences associated with genetic alteration of actin cytoskeletal regulatory genes. These advances have provided new insights into the role of cortical actin cytoskeletal regulation in a number of immune cell functions and have identified cytoskeletal regulatory proteins critical for normal immune system activity and susceptibility to autoimmunity. PMID- 20001426 TI - Examining the roles of cannabinoids in pain and other therapeutic indications: a review. AB - IMPORTANCE OF THE FIELD: In recent times, our knowledge of cannabinoids and the endocannabinoid system has greatly advanced. With expanding knowledge, synthetic cannabinoids - including nabilone, dronabinol and a combination of synthetic Delta9-THC and cannabidiol - have been developed and tested for benefit in a variety of therapeutic indications. AREAS COVERED IN THIS REVIEW: The aim of this article is to provide a summative review of the vast amount of clinical trial data now available on these agents. WHAT THE READER WILL GAIN: To locate clinical trials for review, a literature search was performed using PubMed between the dates of 25 May and 30 June 2009. Search parameters were set to isolate only human randomized controlled trials (RCTs) published between 1990 and 2009. Keywords consistently used for each search include: cannabinoids, marijuana, THC, nabilone and dronabinol. Preferential selection was given to the best-designed trials, focusing on placebo-controlled, double-blind RCTs with the largest patient populations, if available. TAKE HOME MESSAGE: As efficacy and tolerability of these agents remain questionable, it is important that cannabinoids not be considered 'first-line' therapies for conditions for which there are more supported and better-tolerated agents. Instead, these agents could be considered in a situation of treatment failure with standard therapies or as adjunctive agents where appropriate. PMID- 20001425 TI - Treatment strategies for dystonia. AB - IMPORTANCE OF THE FIELD: Dystonia is a neurological syndrome characterized by involuntary twisting movements and unnatural postures. It has many different manifestations and causes, and many different treatment options are available. These options include physical and occupational therapy, oral medications, intramuscular injection of botulinum toxins, and neurosurgical interventions. AREAS COVERED IN THIS REVIEW: In this review, we first summarize the treatment options available, then we provide suggestions from our own experience for how these can be applied in different types of dystonia. In preparing this review article, an extensive literature search was undertaken using PubMed. Only selected references from 1970 to 2008 are cited. WHAT THE READER WILL GAIN: This review is intended to provide the clinician with a practical guide to the treatment of dystonia. TAKE HOME MESSAGE: Treatment of dystonia begins with proper diagnosis and classification, followed by an appropriate search for underlying etiology, and an assessment of the functional impairment associated with the dystonia. The therapeutic approach, which is usually limited to symptomatic therapy, must then be tailored to the individual needs of the patient. PMID- 20001427 TI - Insulin therapy in type 2 diabetes: insulin analogue mix 50, a potential role in reducing postprandial hyperglycaemia and cardiovascular disease. AB - IMPORTANCE OF THE FIELD: Postprandial hyperglycaemia is becoming topical, with studies suggesting a link to cardiovascular disease. Recently, a number of new therapies for the treatment of type 2 diabetes have become available. AREAS COVERED IN THIS REVIEW: This review looks at the evidence for the potential role of insulin analogue mix 50 to reduce postprandial hyperglycaemia and cardiovascular disease. SEARCH STRATEGY: Medline and Embase databases were searched using the MeSH terms to identify relevant studies from 1980 to 2009. Both original articles and reviews were extracted. Published reference lists were also examined. MeSH terms used for literature searching: human insulins, insulin analogues, insulin analogue mix 50, glycaemia, postprandial glucose, fasting glucose, type 2 diabetes, type 1 diabetes, cardiovascular disease. WHAT THE READER WILL GAIN: The reader is presented with evidence discussing the importance of postprandial hyperglycaemia and studies comparing different insulin regimes and in particular insulin analogue mix 50 and its potential to reduce postprandial glucose surges and reduce cardiovascular disease. TAKE-HOME MESSAGE: Insulin analogue mix 50 is a viable therapeutic option in a sub-group of patients with type 2 diabetes. PMID- 20001428 TI - Risk of additional cancers in untreated and treated hairy cell leukemia patients. AB - IMPORTANCE OF THE FIELD: One of the feared events encountered in hairy cell leukemia (HCL) survivors is the subsequent development of a malignant neoplasm. The increased incidence of second cancers in HCL has been documented in large epidemiologic studies conducted in various locations on the globe. AREAS COVERED IN THIS REVIEW: The authors explore the current clinico-epidemiologic evidence, as well as the immune alterations, that link HCL and its therapies to the development of second cancers. Most relevant publications have been identified through the PubMed/Medline database search. WHAT THE READER WILL GAIN: Although HCL patients could develop both HCL and secondary malignancies because of a shared genetic predisposition, a common environmental carcinogen, or not yet identified infectious agents, multiple immune defects documented in HCL might play an important role in second carcinogenesis. Furthermore, the 'gold standards' of HCL therapy - cladribine and pentostatin - are associated with profound and prolonged suppression of the CD4(+) T-lymphocyte counts, often in excess of 2 - 3 years. And while there is no clear-cut evidence that pentostatin or interferon-alpha play an established role in generation of an excess of second cancers in HCL, the safety of cladribine, the preferred agent by a majority of clinicians worldwide, in this regard is a still largely unsettled issue. TAKE HOME MESSAGE: Therefore, it remains to be seen if the immune deficiencies induced by the HCL therapies and their consequences can be offset by the benefit conferred by controlling the leukemic process. PMID- 20001429 TI - HER-2 targeted treatment of osteosarcoma: the challenges of developing targeted therapy and prognostic factors for rare malignancies. AB - IMPORTANCE OF THE FIELD: Despite the substantial success realized treating osteosarcoma with chemotherapy, metastatic disease or refractory/recurrent disease continue to frustrate patients and clinicians. The failure to improve outcomes with treatment intensification indicates the need for new approaches, such as the introduction of targeted therapy. The application of trastuzumab (Herceptin((R))) in the setting of HER-2 positive osteosarcoma is reflective of this concept. AREAS COVERED IN THIS REVIEW: This review summarizes reports from 1996 to the present which focus on HER-2 expression and its prognostic relevance in the setting of osteosarcoma. WHAT THE READER WILL GAIN: The prognostic value of HER-2 remains controversial, and both standard methodologies for assessing expression and the effectiveness of HER-2 directed therapy in this patient population have yet to be established. TAKE HOME MESSAGE: The relative rarity of the disease makes larger prospective studies difficult in terms of both time and coordination. The feasibility of defining therapy specifically for a subpopulation of patients afflicted with a rare cancer, such as osteosarcoma, can be conceptually questioned. PMID- 20001430 TI - Palliative treatment of unresectable metastatic colorectal cancer. AB - IMPORTANCE OF THE FIELD: Treatment options for metastatic colorectal cancer (mCRC) patients have rapidly increased in the past years, but 50 - 70% of mCRC patients are still unlikely to undergo radical resection of metastases and are candidates for palliative therapy only. AREAS COVERED IN THIS REVIEW: Oxaliplatin and irinotecan have widened the chemotherapy alternatives available in this setting and effective targeted agents against vascular endothelial growth factor and epidermal growth factor receptor have further improved treatment efficacy. This review covers the main areas of debate in the optimal treatment of unresectable mCRC patients, focusing on the implications for everyday clinical practice and future research of the most relevant clinical trials and molecular investigations published from 1999 to 2009. WHAT THE READER WILL GAIN: Insights into treatment individualization strategies are provided in the review. TAKE HOME MESSAGE: 'One size fits all' can not longer be considered an adequate approach to unresectable mCRC, and treatment with both chemotherapy and biologic agents should be guided by prognostic and predictive factors in order to maximize the benefit while reducing futile toxicities. PMID- 20001431 TI - Pharmacotherapy of regional melanoma therapy. AB - IMPORTANCE OF THE FIELD: In-transit melanoma metastases develop within regional dermal and subdermal lymphatics before reaching the regional lymph nodes. The prognosis is poor and comparable to multiple nodal metastases. Isolated limb infusion (ILI) or perfusion (ILP) are effective treatments for unresectable, in transit melanoma, with response rates reaching 95%. Although ILI and ILP are more effective than systemic therapy, most patients will recur, thus highlighting the need for newer strategies to improve durable response rates. AREAS COVERED IN THIS REVIEW: We review historical and current literature from 1958 to 2009 regarding regional therapy for melanoma, with focus on the ILI and ILP techniques, pharmacokinetics and resistance mechanisms of melphalan. Alternative therapies, adjunct strategies and new targeted therapies aimed at improving response rates and long-term remission are also discussed. WHAT THE READER WILL GAIN: The reader will gain a comprehensive review on regional pharmacotherapy for melanoma, including alternative therapies, adjunct strategies and new targeted therapies. TAKE HOME MESSAGE: Regional chemotherapy is a viable, evolving treatment for patients with in-transit melanoma and a springboard for ongoing research aimed at improving therapies for malignant melanoma. PMID- 20001432 TI - Diagnosis and treatment of fungal infections in allogeneic stem cell and solid organ transplant recipients. AB - IMPORTANCE OF THE FIELD: Invasive fungal diseases (IFD) are severe complications in patients receiving immunosuppression after solid organ or allogeneic stem cell transplantation. Extensive study has been conducted on therapeutic strategies for IFD in neutropenic patients, mostly those with hematological malignancy. There is an ongoing discussion on whether these studies may be applied to transplant patients as well. AREAS COVERED IN THIS REVIEW: We have reviewed relevant literature on transplantation and clinical mycology of the last 20 years and selected articles relevant for today's treatment decisions. This article reports on the epidemiology of IFD in transplant recipients and current antifungal drugs in the context of tansplantation medicine. For invasive aspergillosis and invasive candidiasis, we give a detailed report of current clinical evidence. WHAT THE READER WILL GAIN: This review is intended as a quick-start for clinicians and other care providers new to transplant care and as an update for experienced transplant physicians. In a field in which evidence is scarce and conflicting, we provide evidence-based strategies for diagnosing and treating the most relevant IFD in transplant recipients. TAKE HOME MESSAGE: Physicians treating transplant patients should maintain a high level of awareness towards IFD. They should know the local epidemiology of IFD to make the optimal decision between current diagnostic and therapeutic strategies. Prophylaxis or early treatment should be considered given the high mortality of IFD. PMID- 20001433 TI - Zonisamide in the treatment of epilepsy. AB - IMPORTANCE OF THE FIELD: More than 1 million epilepsy patients suffer from insufficiently controlled epilepsy, both in the USA and in Europe. Zonisamide is an antiepileptic drug with multiple mechanisms of action, corresponding to efficacy in diverse epilepsy syndromes. AREAS COVERED IN THIS REVIEW: Here, an update on pharmacodynamics, pharmacokinetics, clinical efficacy and safety in childhood and adult epilepsies is given based on an analysis of controlled and uncontrolled studies, European, US and East Asian, and on clinical experience with zonisamide (ZNS) published up to 2009. WHAT THE READER WILL GAIN: Evidence is presented that ZNS is effective not only in adult focal epilepsies, for which it has been approved in the USA and in Europe, but also may offer treatment options for compassionate use in a spectrum of difficult-to-treat epilepsy syndromes. Its favorable pharmacokinetic profile allows for easy combination with most available antiepileptic drugs. TAKE HOME MESSAGE: Provided that additional studies on ZNS are performed to extend approval and to generate comparative data, ZNS has a potential to gain importance in the treatmnt of a wide spectrum of epilepsy patients. PMID- 20001434 TI - Tadalafil for the treatment of pulmonary arterial hypertension. AB - BACKGROUND: Tadalafil, a long-acting phosphodiesterase-5 inhibitor (PDE-5) is the most recent oral agent to receive FDA approval for the treatment of pulmonary arterial hypertension (PAH). OBJECTIVE: With several new agents emerging for the treatment of PAH, this article reviews tadalafil, the compound and its properties, clinical evidence supporting its use, and the role of tadalafil in the current treatment approach for patients with PAH. METHODS: A broad PubMed literature search was performed to identify the most current data on the use of tadalafil for PAH. RESULTS: Tadalafil received FDA approval in 2009 following the recently published pivotal trial that demonstrated that the use of tadalafil 40 mg once daily was well tolerated, improved exercise capacity and quality of life measures and reduced time to clinical worsening in PAH patients. As the second PDE-5 inhibitor to gain approval for PAH, clinical properties such as its long half-life leading to once-daily dosing and possibly improved compliance, as well as potential cost benefit, may distinguish tadalafil from sildenafil in the widespread treatment of PAH. PMID- 20001435 TI - Talaporfin sodium. AB - IMPORTANCE OF THE FIELD: Despite therapeutic advances, cancer remains the cause of an estimated 23% of deaths in the USA. New treatments for malignancy are greatly needed. AREAS COVERED IN THIS REVIEW: Talaporfin sodium is a light activated drug that causes tissue death through induction of apoptosis. Systemic antitumor effects mediated by CD8(+) T cells have been demonstrated in preclinical studies, providing a mechanism for distant response of tumors noted in clinical trials. Talaporfin sodium is approved in Japan for early-stage endobronchial cancer. Phase I and II studies in solid tumors have shown tumor regression in patients refractory to other therapies. Phase III pivotal studies against hepatocellular carcinoma as monotherapy and liver-metastatic colorectal cancer in combination with chemotherapy are ongoing. Talaporfin sodium is also in studies in men with symptomatic benign prostatic hyperplasia. Substantial safety data from clinical trials so far indicate that the drug is well tolerated. WHAT THE READER WILL GAIN: Talaporfin sodium has a broad safety profile and a mode of action that could affect growth in treated and untreated tumors. TAKE HOME MESSAGE: Clinical and preclinical studies indicate that talaporfin sodium treatment may offer a powerful option to synergize current therapies, as well as an alternative monotherapy in treating cancer. PMID- 20001436 TI - Zoledronic acid: an unending tale for an antiresorptive agent. AB - IMPORTANCE OF THE FIELD: Therapy with zoledronic acid (ZOL) is a standard for patients with malignant bone disease. However, the antitumor effects of ZOL are demonstrated only in the preclinical setting and, therefore, they require pharmacological and molecular optimization. ZOL blocks protein isoprenylation, a pleiotropic and basic biochemical process within the cells regulating the activity of several proteins involved in survival and proliferating pathways. AREAS COVERED IN THIS REVIEW: The present review will cover different areas of research from pharmacology to cell biology and molecular biology regarding both direct and indirect anticancer effects of ZOL in humans. The challenge in demonstrating the clinical activity of ZOL is mainly based on its pharmacokinetic properties, which are discussed in detail. WHAT THE READER WILL GAIN: New data about the modes to overcome the limitations of ZOL and the following therapeutic strategies to implement its anticancer activity are described: i) development of pharmacological combinations; ii) finding of new molecular targets of ZOL; and iii) development of new pharmacological formulations of ZOL. TAKE HOME MESSAGE: In conclusion, ZOL has still underestimated anticancer properties that require further investigation in both preclinical and clinical settings. PMID- 20001437 TI - Pharmacotherapeutic efficacy of preservative-free intravitreal triamcinolone acetonide. AB - Preservative-free formulations of triamcinolone acetonide have recently been introduced to the market over concerns of local toxicity of the vehicle and preservatives, including benzyl alcohol in the original formulation, which was not designed for intraocular use. The pharmacokinetics and pharmacodynamics of intravitreal triamcinolone (IVTA) are discussed. The therapeutic effects of IVTA include improvement of visual acuity and reduction of macular edema. However, ongoing treatment is usually required to maintain its effects. The efficacy of IVTA for both FDA-approved and 'off-labeled' indications is reviewed. Elevation of intraocular pressure and cataract formation are the two major side effects of IVTA; these are manageable but require close long-term follow-up. More studies are required to determine the optimal dosage and treatment frequency in different posterior segment disease. PMID- 20001439 TI - Artificial intelligence for diagnostic purposes: principles, procedures and limitations. AB - BACKGROUND: Back propagation (BP) artificial neural networks are a distribution free method for data analysis based on layers of artificial neurons that transduce imputed information. It has been recognized as having a number of advantages compared to traditional methods including the possibility to process imperfect data, and complex non-linear data. The objective of this study was to review the principles, procedures, and limitations of BP artificial neural networks for a non-mathematical readership. METHODS: A real data sample of weight, height and measured body surface area from 90 individuals was used as an example. SPSS 17.0 with neural network add-on was used for the analysis. The predicted body surface from a two hidden layer BP neural network was compared to the body surface calculated by the Haycock equation. RESULTS: Both the predicted values from the neural network and from the Haycock equation were close to the measured values. A linear regression analysis with neural network as predictor produced an r(2)-value of 0.983, while the Haycock equation produced an r(2) value of 0.995 (r(2)>0.95 is a criterion for accurate diagnostic testing). CONCLUSIONS: BP neural networks may, sometimes, predict clinical diagnoses with accuracies similar to those of other methods. However, traditional statistical procedures, such as regression analyses need to be added for testing their accuracies against alternative methods. Nonetheless, BP neural networks have great potential through their ability to learn by example instead of learning by theory. PMID- 20001440 TI - Experimental validation of specificity of the squamous cell carcinoma antigen immunoglobulin M (SCCA-IgM) assay in patients with cirrhosis. AB - BACKGROUND: Squamous cell carcinoma antigen-immunoglobulin M (SCCA-IgM) is a useful biomarker for the risk of development of hepatocellular carcinoma (HCC) in patients with cirrhosis due to its progressive increase associated to HCC evolution. In patients with cirrhosis, other assays have been affected by interfering reactivities of IgM. In this study, the analytical specificity of the SCCA-IgM assay was assessed by evaluating SCCA-IgM measurement dependence on different capture phases, and by measuring the recovery of SCCA-IgM reactivity following serum fractionation. METHODS: Serum samples from 82 patients with cirrhosis were analyzed. SCCA-IgM was measured using the reference test (Hepa-IC, Xeptagen, Italy) that is based on rabbit oligoclonal anti-squamous cell carcinoma antigen (SCCA) and a dedicated ELISA with a mouse monoclonal anti-SCCA as the capture antibody. RESULTS: SCCA-IgM concentrations measured with the reference assay (median value=87 AU/mL) were higher than those measured with the mouse monoclonal test (median value=78 AU/mL). However, the differences in the SCCA-IgM distribution were not statistically significant (p>0.05). When SCCA-IgM concentrations measured with both tests were compared, a linear correlation was found (r=0.77, p<0.05). Fractionation of the most reactive sera by gel-filtration chromatography showed that total recovery of SCCA-IgM reactivity was seen only in the fractions corresponding to components with a molecular weight higher than IgM and SCCA (>2000 kDa) with both tests. CONCLUSIONS: The equivalence of both SCCA IgM assays and the absence of reactivity not related to immune complexes support the analytical specificity of SCCA-IgM measurements. The results validate the assessment of SCCA-IgM for prognostic purposes in patients with cirrhosis. PMID- 20001441 TI - Effect of inflammation induced by prolonged exercise on circulating erythroid progenitors and markers of erythropoiesis. AB - BACKGROUND: Exercise in humans augments the mobilization of circulating hematopoietic progenitor cells (CD34(+)) from the bone marrow. We investigated the effect of inflammation on erythroid marrow activity by mobilization of erythroid progenitor cells (EPs) along with soluble markers of erythropoiesis. METHODS: Ten healthy athletes who participated in an ultradistance foot race participated in the study. Peripheral blood mononuclear cells were isolated, before (phase I), at the end (phase II), and at 48 h post-race (phase III). EPs were detected as burst colony forming units (BFU-e) and colonies were scored at day 14. Markers of inflammation (C-reactive protein, serum amyloid-A, interleukin 6, ferritin and S100B) and bone marrow activity (erythropoietin, soluble transferrin receptor and lipocalin-2) were assessed. RESULTS: An approximately three-fold decrease in BFU-e number was observed at phase II. sTfR concentrations were also decreased at phase II and remained decreased at phase III. However, EPO and lipocalin-2 concentrations reached a maximum value at phase II, with a tendency to decrease at phase III. CONCLUSIONS: These findings indicate that exercise-induced inflammation modulates bone marrow homeostasis leading to an increase in leukocyte turnover and a decrease in erythroid compartment. It appears that lipocalin-2 is the main factor that regulates the production and mobilization of EPs. PMID- 20001442 TI - A novel sensitive immunoassay by nucleic acid barcode dot and its application in the detection of prostate-specific antigen. AB - BACKGROUND: The sensitivity and selectivity of traditional methods limits ultramicro detection of proteins. Bio-barcode amplification detection methods based on nanotechnology enables ultramicro detection of protein. However, bio barcode amplification detection depends on the oligonucleotides being fixed on a glass chip. It also requires specialized equipment, which limits its application. We introduce a nano-nucleic acid barcode dot detection technology to determine ultramicro concentrations of protein. The method is simple, quick and accurate. METHODS: Magnetic probe (IgG-M) and dual-labeled gold nanoparticle bio-probe (IgG Au-DNA) were prepared. Protein was captured using a sandwich assay technique and magnetic separation was used. The DNA barcode was released with dithiothreitol (DTT) and detected directly without the requirement for polymerase chain reaction (PCR). Serum prostate-specific antigen (PSA) from 135 patients was detected with this method and compared with enzyme linked immunosorbent assay (ELISA) and radioimmunoassay (RIA). RESULTS: Each IgG-Au-DNA could be covered with 138+/-47 oligonucleotides and 11+/-3 antibodies. The IgG-M could bind 118 mug of antibody per mg. The sensitivity of nano-nucleic acid barcode dot detection technology might allow detection of 1 fg/mL. There were no significant differences in serum PSA from 135 patients when comparing the three methods (compared with ELISA, r=0.950; and with RIA, r=0.967). CONCLUSIONS: The nucleic acid barcode dot method does not require special equipment or complex procedures, but its detection limit is 2-3 orders of magnitude lower than ELISA. PMID- 20001444 TI - Long-term course of Meniere's disease revisited. AB - CONCLUSIONS: The investigated studies disclose a large variability of the many symptoms appearing in the course of Meniere's disease. OBJECTIVES: To analyze the data of 46 mostly retrospective studies (with a total of 7852 patients). METHODS: We present a short overview of the literature on the long-term course of Meniere's disease. Special emphasis is placed on the most relevant and distressing symptoms of these patients, such as vertigo attacks, vestibular and hearing function, drop attacks, and the probability of transition from unilateral to bilateral involvement. RESULTS: The frequency of vertigo attacks diminishes within 5-10 years. Hearing loss (of about 50-60 dB) and vestibular function decrement (of about 35-50%) take place mainly in the first 5-10 years of disease. Drop attacks may occur early or late in the course of the disease, and remission is spontaneous in cmost cases. Bilaterality of the condition increases with increasing duration of the disease (up to 35% within 10 years, up to 47% within 20 years). PMID- 20001445 TI - Short-term endotracheal climate changes and clinical effects of a heat and moisture exchanger with an integrated electrostatic virus and bacterial filter developed for laryngectomized individuals. AB - CONCLUSION: Both the regularly used heat and moisture exchanger (R-HME) and the HME with both an antimicrobial and hygroscopic element (F-HME) are effective moisture exchangers. The antimicrobial filter of the F-HME acts as a heat exchanger. The external features of the F-HME were experienced as inconvenient, but decreased sputum production was reported as well. OBJECTIVES: Recently an HME with an integrated antimicrobial filter has become available for use in laryngectomized patients. The purpose of this study was to assess its short-term endotracheal climate changes and feasibility in daily practice. METHODS: Endotracheal temperature and humidity were successfully measured in 13 laryngectomized patients (2652 analysed full breaths), during 10 min rest breathing with the R-HME, with an F-HME and without HME in a randomized sequence. Additionally, a 3 week prospective clinical feasibility trial was conducted in 17 laryngectomized patients. RESULTS: Both R-HME and F-HME increase endotracheal minimum humidity values (5.8 and 4.7 mgH(2)O/L, respectively; p < 0.0001). Compared with open stoma breathing, in contrast to the R-HME, the F-HME increases both end-inspiratory and end-expiratory temperature values (1.1 degrees C, and 0.6 degrees C, respectively). After the 3-week clinical feasibility trial, one patient dropped out; 11 patients (11/16 = 69%) disliked the larger design of the F-HME and all patients reported less optimal airtight occlusion. Five patients (5/16 = 31%) reported remarkably decreased sputum production. PMID- 20001446 TI - Role of spinning top urethra in dysfunctional voiding. AB - OBJECTIVE: The role of spinning top urethra (STU) in children with dysfunctional voiding was evaluated retrospectively. MATERIAL AND METHODS: From 1995 to 2002, the records of 154 children with dysfunctional voiding were reviewed retrospectively. Of the children 110 (71%) were girls and 44 (29%) were boys (mean age 8 years, range 4-14). All children were neurologically normal and no exhibited physical signs of occult spinal dysraphism. Patients were divided into two groups according to their width of proximal urethra: group I had STU and the group II had normal urethral width. The groups were compared with each other for gender, voiding symptoms, urinary tract infection (UTI), vesicoureteral reflux (VUR) and urodynamic observations. RESULTS: There were 84 children (mean age 8.3 +/- 2.2 years, range 4-14) in group I and 70 (mean age 8.0 +/- 2.1 years, range 4 14) in group II; no significant age difference was found between the two groups (p = 0.4674). Group I consisted of 66 (71%) girls and 18 (29%) boys and group II 44 (63%) girls and 26 (37%) boys. STU was observed more in girls than boys in group I (p = 0.0316). UTI was observed in 57 patients (68%) in group I and 34 (49%) in group II (p = 0.0154). Mean duration of symptoms was 42 +/- 24 months (range 6-118) and 39 +/- 23 (range 3-120) months in groups I and II, respectively (p = 0.6302). Postvoid residual urine (PVR) more than 10% of expected bladder capacity was detected in 15 patients (18%) in group I and seven (10%) in group II. No association was found between the meaningful PVR and STU (p = 0.1653). The presence of detrusor overactivity during filling was observed in 54 patients (64%) in group I and 42 (60%) in group II (p = 0.4676). Diminished bladder compliance (< 10 ml/cmH(2)O) was detected in 34 patients (40%) in group I and 17 (24%) in group II (p = 0.0335). The mean voiding pressure was measured as 56 +/- 29 cmH(2)O in group I, which was significantly higher than in group II (49 +/- 25 cmH(2)O) (p = 0.0373). The mean flow rate during the emptying phase of urodynamics was 16 +/- 8 and 15 +/- 6 ml/s in groups I and II, respectively (not significant, p = 0.2686). VUR was detected in 16 patients (19%) in group I and two (3%) in group II (p = 0.0018). CONCLUSIONS: STU was related to recurrent UTIs, VUR, poor bladder compliance and more serious functional urinary obstruction. Furthermore, STU may be a consequence of a neurogenic maturation defect in detrusor-sphincter coordination resembling that of urofacial syndrome, because development of this situation was found to be independent of the duration of symptoms. PMID- 20001447 TI - Music exposure and hearing disorders: an overview. AB - It has been generally accepted that excessive exposure to loud music causes various hearing symptoms (e.g. tinnitus) and consequently leads to a risk of permanent hearing damage, known as noise-induced hearing loss (NIHL). Such potential risk of NIHL due to loud music exposure has been widely investigated in musicians and people working in music venues. With advancements in sound technology and rapid developments in the music industry, increasing numbers of people, particularly adolescents and young adults, are exposing themselves to music on a voluntary basis at potentially harmful levels, and over a substantial period of time, which can also cause NIHL. However, because of insufficient audiometric evidence of hearing loss caused purely by music exposure, there is still disagreement and speculation about the risk of hearing loss from music exposure alone. Many studies have suggested using advanced audiological measurements as more sensitive and efficient tools to monitor hearing status as early indicators of cochlear dysfunction. The purpose of this review is to provide further insight into the potential risk of hearing loss caused by exposure to loud music, and thus contribute to further raising awareness of music induced hearing loss. PMID- 20001448 TI - Mercury, dental amalgam, and hearing loss. PMID- 20001449 TI - Where are teachers' voices? A research agenda to enhance the communicative interactions of students with multiple and severe disabilities at school. AB - PURPOSE: Despite the importance of communication for educational participation, researchers have observed low frequencies of communication between teachers and students with multiple and severe disabilities (MSD). Little is known about reasons for these low frequencies. This article explores the literature to develop a new research agenda for approaching this issue. METHOD: Selected studies were reviewed, including qualitative studies involving teachers of students who use augmentative and alternative communication (AAC) in mainstream classrooms. RESULTS: Teachers identified a broad range of issues that influence their ability to educate students who use AAC or demonstrate other significant communication needs. These factors include teachers' attitudes, perceptions and beliefs and broader contextual factors, such as administrative support at the institutional level. However, to date, there has been little research with teachers of students with MSD about factors influencing communication with their students. CONCLUSIONS: A broad range of potential factors may impact upon teachers' ability to communicate with their students. Further research involving teachers of students with MSD is needed to explore the range of potential factors influencing low frequencies of communication. This research may contribute to an understanding of how best to support teachers to enhance communication access for students with MSD in the classroom. PMID- 20001450 TI - Population pharmacokinetics of posaconazole in neutropenic patients receiving chemotherapy for acute myelogenous leukemia or myelodysplastic syndrome. AB - OBJECTIVE: The relationship between patient characteristics and posaconazole exposures was evaluated in a population pharmacokinetic (PK) model using trial data from neutropenic patients administered oral posaconazole suspension as antifungal prophylaxis. METHODS: Data were analyzed using nonlinear mixed-effects modeling. Covariates were tested using the forward addition, Objective Function (OF) cut-off of 3.84, followed by the backward elimination (OF cut-off 10.88) steps in NONMEM. These covariates included demographics, mucositis, neutropenia, vomiting, diarrhea, proton pump inhibitor (PPI) or H(2)-receptor antagonist usage and baseline bilirubin or baseline gamma-glutamyl transferase (GGT) levels > or =2 x upper limit of normal (ULN). A correlation between posaconazole PK and the occurrence of invasive fungal infection (IFI) was also examined. RESULTS: Statistically significant associations were demonstrated between posaconazole PK and diarrhea, PPI intake, race, and baseline GGT and bilirubin levels. These covariates did not predominate in patients who developed IFI. CONCLUSION: This analysis provides information regarding the correlation of patient covariates with posaconazole exposures estimated in a clinical setting. The results of this analysis agree with previously reported analyses. However, because of the successful prophylaxis and the low number of posaconazole-treated patients with IFI proven or probable (IFIPP), the absence of a statistically significant relationship between IFIPP and exposure may not mean this relationship does not exist. A meta-analysis of several efficacy trials or exploring alternate composite endpoints for efficacy may be needed to answer this question. PMID- 20001451 TI - Cost-effectiveness of single agent, uptitration and switching statin treatment strategies for lipid lowering in Sweden. AB - OBJECTIVES: To assess which alternative treatment strategies are optimum in terms of cost-effectiveness (EUR/patient treated to target, EUR/PTT) in lowering cholesterol in high-risk patients with elevated LDL-cholesterol (LDL-C) in Sweden. METHODS: A probabilistic cost-effectiveness model was developed to estimate the mean expected costs and proportion of patients reaching goal attainment (defined as LDL-C < or =2.5 mmol/L [96.5 mg/dL]) at some point in time within a 52-week period following the initiation of statin therapy. Eight different statin treatment strategies were evaluated. Key data sources used in the modeling were the scientific literature, hospital tariffs and medicine price databases. RESULTS: Depending on baseline LDL-C and the willingness-to-pay per additional PTT, the cost-effective alternative is always found among four out of the eight assessed treatment strategies (i.e. Simva10 --> Simva20 --> Simva40, Rosu10, Simva20 --> Rosu10 --> Rosu20 --> Rosu40, or Simva20 --> Simva40 --> Rosu20 --> Rosu40). An important finding was that when LDL-C level exceed 4.0 mmol/L (154mg/dL) and when willingness to pay is less than 500 EUR per additional PTT, the optimal treatment strategy would be to initiate cholesterol-lowering treatment directly with rosuvastatin 10 mg. CONCLUSIONS: The results of this study indicate that the optimal approach to initiate lipid-lowering therapy would be to treat patients with the lower baseline LDL-C levels with the least costly treatment strategies, while initiating lipid-lowering treatment with a high potency statin (rosuvastatin) in patients with moderately high or high baseline LDL-C levels. This recommendation can be assumed to be relevant particularly when the fact that after treatment initiation the majority of Swedish patients will not have any changes in their lipid-lowering medication or dose is taken into account. Finally, since only the short-term results are presented here, it would be valuable to conduct further studies of the long-term cost-effectiveness of different statin treatment strategies that focus on treatment persistence and LDL C goal attainment in real practice. PMID- 20001452 TI - Clinical adenoviral gene therapy for prostate cancer. AB - Prostate cancer is at present the most common malignancy in men in the Western world. When localized to the prostate, this disease can be treated by curative therapy such as surgery and radiotherapy. However, a substantial number of patients experience a recurrence, resulting in spreading of tumor cells to other parts of the body. In this advanced stage of the disease only palliative treatment is available. Therefore, there is a clear clinical need for new treatment modalities that can, on the one hand, enhance the cure rate of primary therapy for localized prostate cancer and, on the other hand, improve the treatment of metastasized disease. Gene therapy is now being explored in the clinic as a treatment option for the various stages of prostate cancer. Current clinical experiences are based predominantly on trials with adenoviral vectors. As the first of a trilogy of reviews on the state of the art and future prospects of gene therapy in prostate cancer, this review focuses on the clinical experiences and progress of adenovirus-mediated gene therapy for this disease. PMID- 20001454 TI - Development of lentiviral vectors with optimized transcriptional activity for the gene therapy of patients with Fanconi anemia. AB - Fanconi anemia (FA) is an inherited genetic disease characterized mainly by bone marrow failure and cancer predisposition. Although gene therapy may constitute a good therapeutic option for many patients with FA, none of the clinical trials so far developed has improved the clinical status of these patients. We have proposed strategies for the genetic correction of bone marrow grafts from patients with FA, using lentiviral vectors (LVs). Here we investigate the relevance of the expression of FANCA to confer a therapeutic effect in cells from patients with FA-A, the most frequent complementation group in FA. Our data show that relatively weak promoters such as the vav or phosphoglycerate kinase (PGK) promoter confer, per copy of FANCA, physiological levels of FANCA mRNA in lymphoblastoid cell lines, whereas the cytomegalovirus and, more significantly, spleen focus-forming virus (SFFV) promoters mediated the expression of supraphysiological levels of FANCA mRNA. Insertion of the woodchuck hepatitis virus posttranscriptional regulatory element (WPRE) or a mutated WPRE into the 3' region of PGK-FANCA LVs significantly increased FANCA mRNA levels. At the protein level, however, all tested vectors conferred, per copy of FANCA, similar and physiological levels of the protein, except SFFV LVs, which again conferred supraphysiological levels of FANCA. In spite of their different activity, all tested vectors mediated a similar phenotypic correction in FA-A lymphoblastoid cell lines and also in hematopoietic progenitors from patients with FA-A. On the basis of the efficacy and safety properties of PGK LVs, a PGK LV carrying FANCA and a mutant WPRE is proposed as an optimized vector for the gene therapy of patients with FA-A. PMID- 20001453 TI - Non-cardiomyocytes influence the electrophysiological maturation of human embryonic stem cell-derived cardiomyocytes during differentiation. AB - Various types of cardiomyocytes undergo changes in automaticity and electrical properties during fetal heart development. Human embryonic stem cell-derived cardiomyocytes (hESC-CMs), like fetal cardiomyocytes, are electrophysiologically immature and exhibit automaticity. We used hESC-CMs to investigate developmental changes in mechanisms of automaticity and to determine whether electrophysiological maturation is driven by an intrinsic developmental clock and/or is regulated by interactions with non-cardiomyocytes in embryoid bodies (EBs). We isolated pure populations of hESC-CMs from EBs by lentivirus-engineered Puromycin resistance at various stages of differentiation. Using pharmacological agents, calcium (Ca(2+)) imaging, and intracellular recording techniques, we found that intracellular Ca(2+)-cycling mechanisms developed early and contributed to dominant automaticity throughout hESC-CM differentiation. Sarcolemmal ion channels evolved later upon further differentiation within EBs and played an increasing role in controlling automaticity and electrophysiological properties of hESC-CMs. In contrast to the development of intracellular Ca(2+)-handling proteins, ion channel development and electrophysiological maturation of hESC-CMs did not occur when hESC-CMs were isolated from EBs early and maintained in culture without further interaction with non-cardiomyocytes. Adding back non-cardiomyocytes to early-isolated hESC CMs rescued the arrest of electrophysiological maturation, indicating that non cardiomyocytes in EBs drive electrophysiological maturation of early hESC-CMs. Non-cardiomyocytes in EBs contain most cell types present in the embryonic heart that are known to influence early cardiac development. Our study is the first to demonstrate that non-cardiomyocytes influence electrophysiological maturation of early hESC-CMs in cultures. Defining the nature of these extrinsic signals will aid in the directed maturation of immature hESC-CMs to mitigate arrhythmogenic risks of cell-based therapies. PMID- 20001455 TI - The mechanism of distinct diurnal variations of renin-angiotensin system in aorta and heart of spontaneously hypertensive rats. AB - Diurnal variations in plasminogen activator inhibitor-1 mRNA expression are different between the spontaneously hypertensive rats (SHRs) and the Wistar-Kyoto (WKY) rats, and between the aorta and the heart. To elucidate the mechanisms, we examined diurnal changes in the circulating renin-angiotensin system in the SHR and WKY rats. Diurnal variations in plasma renin activity (PRA), plasma angiotensin I, and aldosterone concentrations were similar between the SHR and WKY rats. On the other hand, plasma angiotensin II (Ang II) concentration in the SHR was lower than that in the WKY rats at most time points, but increased to the level of the WKY rats in the late light phase. Treatment with AT1 receptor antagonist candesartan increased plasma Ang II concentration except at ZT 8 and lessened its diurnal variation in the SHR. At the peak in plasma Ang II in the SHR, Ang II regulated genes such as transforming growth factor-beta1 and p22phox were upregulated in the aorta. On the other hand, these genes were upregulated throughout the day in the heart of SHR. Candesartan treatment increased AT1a receptor mRNA expression in the heart but not in the aorta of SHR. These findings suggest that an AT1 receptor-mediated mechanism might cause a surge in plasma Ang II concentration at the late light phase in the SHR. Homologous down-regulation of AT1a receptor by Ang II may dampen the effect of a surge in plasma Ang II concentration in the heart of SHR. PMID- 20001456 TI - Does decreased diastolic blood pressure associate with increased mortality in 80 year-old Japanese? AB - Hypertension is one of the greatest risk factors for cardiovascular disease, but its contribution to cardiovascular mortality weakens with aging. We have previously demonstrated that at the age of 80, higher systolic blood pressure (SBP) is not correlated with increased mortality in Japan. However, we did not examine in detail whether diastolic blood pressure (DBP) independently affects mortality. In the present study, 639 participants, who were 80 years old in 1997, were enrolled. The subjects were divided by their DBP [below 70 mmHg (group 1, n = 136), from 70 mmHg to 80 mmHg (group 2, n = 200), from 80 mmHg to 90 mmHg (group 3, n = 194), over 90 mmHg (group 4, n = 109)]. During the 4-year follow-up period, 90 individuals died. Cox multivariate regression analysis revealed that group 1 showed a significantly higher mortality rate than group 4 [relative risk (RR) 2.47, confidence interval (CI) 1.07-5.70, p = 0.03)]. The relative risks of deaths from cardiovascular diseases, pneumonia, and cancer tended to be higher in group 1 than in group 4, but the difference did not reach statistical significance. These results suggest that decreased DBP is associated with higher mortality in the Japanese elderly. PMID- 20001457 TI - Comparison of effects of low dose of spironolactone and a thiazide diuretic in patients with hypertension treated with an angiotensin-converting enzyme inhibitor or an angiotensin type 1 receptor blocker. AB - This study was performed to investigate the additional anti-hypertensive effects and safety of low-dose thiazide diuretic, trichlormethiazide (TCTZ), and a mineralocorticoid receptor blocker, spironolactone (SPI), as add-on therapy in 64 patients whose blood pressure (BP) at office were over 140/90 mmHg, while receiving anti-hypertensive medication including an angiotensin-converting enzyme inhibitor or angiotensin II type I receptor antagonist. After 6 months, we observed a decrease of office and home BP. Moreover, urinary albumin excretion (UAE) was reduced in SPI-treated group, but not in the TCTZ-treated group. No significant change in serum potassium, lipids, glucose, or uric acid was observed. In conclusion, low-dose thiazide diuretic or SPI provided a significant additional anti-hypertensive effect in patients in whom hypertension was not controlled by medication, and SPI-reduced UAE. PMID- 20001458 TI - Zigzagged augmentation index in diabetes. AB - Although the patients with diabetic nephropathy suffered high cardiovascular risk, augmentation index (AI) in diabetic nephropathy has been poorly characterized. Cross-sectional studies were performed on 26 diabetic and 27 nondiabetic nephropathic patients. Home blood pressure was examined. In addition, blood pressure, pulse rate, and AI were measured in both supine and sitting positions. Patient backgrounds such as age, sex, sitting blood pressure, and pulse rate were similar between two groups. Circadian variations of home blood pressure were preserved in nondiabetic patients, but disappeared in diabetes. Changing from supine to sitting position induced greater decrements of systolic blood pressure (DeltaSBP -9 +/- 8 mmHg) and AI (DeltaAI -7 +/- 10) in the diabetic group than in nondiabetic patients (DeltaSBP -4 +/- 12 mmHg, DeltaAI -2 +/- 9). Multivariate regression analysis revealed that AI in a sitting position correlated positively to SBP and inversely to pulse rate. Of interest, AI in supine position related positively to age, the presence of diabetes and SBP, and inversely to pulse rate. The present data indicate autonomic dysfunction in patients with diabetic nephropathy. Furthermore, our findings provide the evidence that autonomic dysfunction elicits an inadequate physiological arterial contraction in response to postural change, thereby reducing AI that results in the fall of SBP. Finally, the present results suggest that AI in supine, but not sitting position, is suited for detecting cardiovascular risk in diabetes. PMID- 20001459 TI - Blood pressure variability as well as blood pressure level is important for left ventricular hypertrophy and brachial-ankle pulse wave velocity in hypertensives. AB - Blood pressure (BP) variability is calculated as the standard deviation of ambulatory BP. Blood pressure variability is associated with the cardiovascular morbidity; however whether it is related to target organ damage is controversial. In this study we examined a possible relationship between the BP variability and left ventricular hypertrophy (LVH), and between BP variability and brachial-ankle pulse wave velocity (baPWV). The present study was conducted on 111 consecutive Japanese hypertensive patients who were hospitalized for the educational program in our hospital under stable sodium chloride intake (6 g/day). Blood pressure measurements were at 30-minute intervals all day. In a multivariable analysis adjusted with confounding factor, LVH was associated with 24-hour systolic BP (SBP), 24 hour diastolic BP (DBP), daytime SBP, daytime DBP, nighttime SBP, and nighttime DBP. Additionally, nighttime DBP variability was related to LVH. By the same method, baPWV as a dependent variable was related to 24-hour SBP and nighttime SBP. Furthermore, nighttime SBP variability was concerned with baPWV. The LVH was associated with not only BP level but also with nighttime DBP variability. The baPWV was also related not only to BP level but also to nighttime SBP variability. PMID- 20001460 TI - Effects of angiotensin II receptor blockers on the relationships between ambulatory blood pressure and anti-hypertensive effects, autonomic function, and health-related quality of life. AB - The aim of the present study was to examine the relationships between the anti hypertensive effects, autonomic function, and health-related quality of life (HRQOL) following treatment of hypertensive subjects with angiotensin receptor blockers (ARBs) in hypertensives. Nineteen patients with hypertension were assigned randomly to daily treatment with ARBs. After 16 weeks of treatment, blood pressure (BP) and 24 h the ratio of low frequency to high frequency component (LF/HF), an index of sympathovagal balance were decreased by ARBs. The HRQOL scores improved during the study. In this study, ARB therapy was associated with an improvement in BP, autonomic function, and HRQOL. PMID- 20001461 TI - Usefulness of self-monitoring of urinary salt excretion in hypertensive patients. AB - We investigated the usefulness of measuring urinary salt excretion by using a self-monitoring device. Subjects were 34 hypertensive patients who underwent successful 24-h home urine collection five times and 25 volunteers. Four volunteers were diagnosed as having hypertension based on home blood pressure (BP) readings. All subjects were asked to measure daily urinary salt excretion for 30 days by using a self-monitoring device which estimates 24-h salt excretion by overnight urine. The mean urinary salt excretion during the 30 days was 8.36 +/- 1.52 g/day and the range (maximum-minimum value) was 5.47 +/- 20.05 g/day in all subjects. Mean urinary salt excretion decreased from 8.52 +/- 1.63 g/day for the first 10 days to 8.31 +/- 1.54 g/day for the last 10 days (p < 0.05). The mean urinary salt excretion determined by a self-monitoring device using overnight urine was positively associated with that determined by 24-h home urine for five times in the hypertensive subjects (r = 0.63, p < 0.01). Results indicate that a self-monitoring device seems to be useful to monitor daily salt intake and to guide salt restriction. PMID- 20001462 TI - Atorvastatin improves the impaired baroreflex sensitivity via anti-oxidant effect in the rostral ventrolateral medulla of SHRSP. AB - We have demonstrated that oxidative stress in the rostral ventrolateral medulla (RVLM), a vasomotor center in brainstem, increases sympathetic nerve activity (SNA) and that oral administration of atorvastatin inhibited SNA via anti-oxidant effect in the RVLM of stroke-prone spontaneously hypertensive rats (SHRSPs). The impairment of baroreflex sensitivity (BRS) is known as the predictive factor of mortality in the hypertension and BRS is impaired in SHRSP. The aim of the present study was to determine whether oral administration of atorvastatin improved the impaired BRS via anti-oxidant effect in the RVLM in SHRSP. Atorvastatin (20 mg/kg/day) or vehicle was orally administered for 28 days in SHRSPs. Systolic blood pressure (SBP), heart rate, and 24-h urinary norepinephrine excretion as an indicator of SNA were comparable between atorvastatin- and control-SHRSP. Thiobarbituric acid-reactive substance (TBARS) levels as a marker of oxidative stress was significantly lower in atorvastatin SHRSP than in control-SHRSP. Baroreflex sensitivity measured by the spontaneous sequence method was significantly higher in atorvastatin-SHRSP than in control SHRSP. These results suggest that atorvastatin improves the impaired BRS in SHRSP via its anti-oxidant effect in the RVLM of SHRSP. PMID- 20001463 TI - Factors related to elevated 24-h blood pressure in young adults. AB - Variation in 24-h blood pressure (BP) is associated with multiple factors, but the association has not been thoroughly examined in young adults. To elucidate the potential factors associated with variation in 24-h BP, 331 healthy medical students were investigated. Awake mean BP negatively correlated with sleep duration in males. Sixty-seven subjects (20.2%) had a high 24-h BP according to the ESH/ESC 2007 guidelines (systolic blood pressure (SBP) 125 and/or diastolic blood pressure (DBP) 80 mmHg). After multivariate analysis for confounding factors, male gender, body mass index (BMI), smoking, the 24-h low/high frequency component (heart rate variability spectral analysis), and short sleep (5 h or less) were found to be associated with high BP. The present study is the first to demonstrate the multivariate risk factors for elevated 24-h BP in a large number of young adults. Further investigation is required to determine the causal relationship between modifiable BP-related factors and elevated 24-h BP in young adults. PMID- 20001468 TI - Sildenafil versus continuous positive airway pressure for erectile dysfunction in men with obstructive sleep apnea: a meta-analysis. AB - OBJECTIVE: To evaluate the effectiveness of sildenafil versus continuous positive airway pressure (CPAP) for patients with erectile dysfunction (ED) and obstructive sleep apnea (OSA). METHODS: This is a meta-analysis of a randomized controlled trial. The main outcome measures for effectiveness were the percentage of successful intercourse attempts, International Index of Erectile Function (IIEF) domain scores (erectile function, EF) and the satisfaction levels of the patients and their partners with the treatment for ED. RESULTS: Two randomized controlled trials totaling 70 patients were included. Meta-analysis results are as follows: after 12 weeks of treatment, patients under sildenafil demonstrated a significant advantage over under CPAP in terms of the percentage of successful intercourse attempts [OR = 3.24, 95% CI (2.37-4.43)], EF scores [WMD = 3.57, 95%CI (1.68-5.45)], and the satisfaction levels of the patients and their partners with the treatment for ED [OR = 3.56, 95% CI (1.27-9.98)]. CONCLUSION: Current clinical studies might confirm that both therapeutic methods were safe and effective, but sildenafil was superior to CPAP in the treatment of ED in men with OSA. We conclude that new therapeutic agents or a combination of the two methods should be studied further. PMID- 20001469 TI - Solifenacin treatment in men with overactive bladder: effects on symptoms and patient-reported outcomes. AB - OBJECTIVE: In the few studies to evaluate antimuscarinics for overactive bladder (OAB) in men, safety has been the primary focus. In OAB efficacy trials, subject populations have been predominantly female and patient-reported outcomes (PROs) have been assessed only recently. We present a post hoc analysis of PRO-based findings from the subset of men (without presumed bladder outlet obstruction [BOO]) from two large, independent, open-label trials of solifenacin. METHODS: Subjects with OAB for > or =3 months received flexibly dosed solifenacin for 12 weeks. At baseline and 4-week intervals, subjects completed the Patient Perception of Bladder Condition (PPBC) and Overactive Bladder Questionnaire (OAB q). In one study, subjects also completed 3-day bladder diaries. At baseline, mean PPBC scores were similar in both studies and indicative of moderate-to severe problems. RESULTS: After 12 weeks of solifenacin, mean PPBC scores improved significantly (p < 0.0001); values were suggestive of minor-to-moderate problems. Mean scores on the OAB-q were also significantly improved after solifenacin (p values < or =0.001). In men without presumed BOO, solifenacin significantly improved PRO measures of symptom bother, health-related quality of life, and overall perception of bladder problems. CONCLUSION: Results from these two studies support the use of solifenacin as a well-tolerated and efficacious treatment option for providing symptom relief in men with OAB without BOO. PMID- 20001470 TI - Elevated high sensitivity C-reactive protein levels in aging men with low testosterone. AB - OBJECTIVE: We examined baseline data from a lipid treatment study to assess the relationship between testosterone (T) and the cardiovascular inflammatory marker, high sensitivity C-reactive protein (hsCRP). METHODS: The baseline T, hsCRP, lipid, glycemic, and anthropometric data were obtained from 467 men (mean age: 52 years). Inclusion criteria included low-density lipoprotein cholesterol > or = 3.4 to 4.9 mmol/l and triglycerides < or = 4.0 mmol/l. The baseline hsCRP levels were examined across the following T subgroups: <6.9 nmol/l (moderate to severe hypogonadism), 6.9 to <10.4 nmol/l (mild to moderate hypogonadism), 10.4 to <15 nmol/l (low-normal T), and > or = 15 nmol/l (normal T). RESULTS: The median hsCRP levels were significantly (p = 0.041) different across the four T subgroups; patients in the lower T subgroups had higher median hsCRP levels than patients in the higher T subgroups. The percentage of men with elevated hsCRP (>2 mg/l) was also significantly (p = 0.038) different across the four T subgroups; 83% of men with T < 6.9 nmol/l had elevated hsCRP compared with 40% with T > or = 15 nmol/l. CONCLUSIONS: This analysis demonstrated an inverse relationship between serum T and hsCRP in aging men. Urologists need to be aware that low T levels may not only adversely affect sexual function but also may worsen cardiovascular risk in aging, hypogonadal men. PMID- 20001471 TI - Effect of clenching on T2 and diffusion parameters of the masseter muscle. AB - BACKGROUND: Persistent muscle contractions during clenching are considered to be one reason for temporomandibular disorders. However, no report has evaluated the effect of clenching on the masticatory muscles, as measured by magnetic resonance imaging (MRI). PURPOSE: To investigate whether clenching has an effect on either T(2) or the coefficients for diffusion of the masseter muscles (MM), and to evaluate the effect of the distribution of bite force on such indices. MATERIAL AND METHODS: Twenty-three subjects were examined. Bite force was measured by a pressure-sensitive sheet, and the force of the right and the left sides was calculated. MRI was used to evaluate T(2), the apparent diffusion coefficient (ADC), and the primary (lambda(1)), secondary (lambda(2)), and tertiary eigenvalues (lambda(3)). These indices on the stronger side of the bite force were compared to those on the weaker side. Thereafter, the indices were compared between at rest and during clenching. RESULTS: There was no significant difference in any of the indices (T(2), ADC, lambda(1), lambda(2), and lambda(3)) between the side of stronger bite force and the side with weaker. T(2) increased by clenching, and the difference was significant in the side with stronger bite force (P = 0.006). ADC, lambda(1), lambda(2), and lambda(3) increased significantly by clenching (P <0.01, P <0.01, P <0.01, and P <0.01, respectively) on both sides. The percentage of change of lambda(2) by clenching was 26.2+/ 15.7% on the stronger side and 26.9+/-18.6% on the weaker side, which was significantly greater than either that of lambda(1) or lambda(3). CONCLUSION: The coefficients for diffusion of the MM were sensitive to change by clenching, and lambda(2) was the most sensitive. Moreover, the relative distribution of the bite forces had no effect on any of the indices. PMID- 20001472 TI - Magnetic resonance imaging signal reduction may precede volume loss in the pituitary gland of transfusion-dependent beta-thalassemic patients. AB - BACKGROUND: Pituitary iron overload in patients with transfusion-dependent beta thalassemia may lead to delayed puberty. Magnetic resonance imaging (MRI) has the potential to estimate tissue iron concentration by detecting its paramagnetic effect and hypophyseal damage by measuring its dimensions indirectly. PURPOSE: To investigate the association of pituitary MRI findings and pubertal status in thalassemic patients as well as to demonstrate any priority in appearance of them. MATERIAL AND METHODS: Twenty-seven beta-thalassemic patients, aged 15-25 years, were divided into 13 with (group A) and 14 without hypogonadism (group B), matched by age, gender, duration of transfusion, and chelation therapy. Thirty eight age- and sex-adjusted healthy control individuals were also included (group C). All participants underwent pituitary MRI using a 1.5 T unit. Pituitary-to-fat signal intensity ratios (SIR) were calculated from coronal T2-weighted images. Estimated pituitary volumes were measured using pituitary height, width, and length on T1-weighted images. RESULTS: The mean values of pituitary-to-fat SIRs were significantly lower in group A as compared with group B (P <0.001), and likewise group B had statistically lower values than group C (P=0.03). The pituitary height and volume were significantly decreased in group A compared to group B (P = 0.006 and P = 0.002, respectively), while these differences did not demonstrate statistically significance between groups B and C. CONCLUSION: Pituitary MRI findings such as signal intensity reduction and decrease in volume can be useful markers in estimating pituitary dysfunction in beta-thalassemic patients. Compared to healthy controls, lower values of pituitary-to-fat SIRs in thalassemic patients experiencing normal puberty, without marked decrease in volume, indicate that signal reduction may precede volume loss and could be expected first on MRI. PMID- 20001473 TI - Galactography: an application of the Galactogram Imaging Classification System (GICS). AB - BACKGROUND: Galactography is the technique of choice for investigating pathological nipple discharge. However, there is no standardized interpretation system for this procedure. PURPOSE: To evaluate radiological and histological correlation using a Galactogram Imaging Classification System (GICS). MATERIAL AND METHODS: We retrospectively studied all galactograms obtained in 62 women with pathological nipple discharge who subsequently had biopsy at our institution between 1999 and 2007. The GICS proposes five categories for galactographic findings: GICS 1: negative; GICS 2: benign; GICS 3: probably benign; GICS 4: suspect for malignancy; GICS 5: highly suspect for malignancy. RESULTS: The galactographic findings were classified as follows: GICS 2: 25.8% (16/62); GICS 3: 30.6% (19/62); GICS 4: 25.8% (16/62); and GICS 5: 17.7% (11/62). A good correlation was observed between histological diagnosis and GICS categories (P < 0.05). All the cases diagnosed with carcinoma (n = 11) were classified in GICS category 5. CONCLUSION: The present study shows a good correlation between GICS categories and histological diagnosis. PMID- 20001474 TI - Magnetic resonance arthrography including ABER view in diagnosing partial thickness tears of the rotator cuff: accuracy, and inter- and intra-observer agreements. AB - BACKGROUND: Partial-thickness tear of the rotator cuff is a common cause of shoulder pain. Magnetic resonance (MR) arthrography has been described as a useful measure to diagnose rotator cuff abnormalities. PURPOSE: To determine the reliability and accuracy of MR arthrography with abduction and external rotation (ABER) view for the diagnosis of partial-thickness tears of the rotator cuff. MATERIAL AND METHODS: Among patients who underwent MR arthrographies, 22 patients (12 men, 10 women; mean age 45 years) who had either partial-thickness tear or normal tendon on arthroscopy were included. MR images were independently scored by two observers for partial-thickness tears of the rotator cuff. Interobserver and intraobserver agreements for detection of partial-thickness tears of the rotator cuff were calculated by using kappa coefficients. The differences in areas under the receiver operating characteristic (ROC) curves were assessed with a univariate Z-score test. Differences in sensitivity and specificity for interpretations based on different imaging series were tested for significance using the McNemar statistic. RESULTS: Sensitivity, specificity, and accuracy of each reader on MR imaging without ABER view were 83%, 90%, and 86%, and 83%, 80%, and 82%, respectively, whereas on overall interpretation including ABER view, the sensitivity, specificity, and accuracy of each reader were 92%, 70%, and 82%, and 92%, 80%, and 86%, respectively. Including ABER view, interobserver agreement for partial-thickness tear increased from kappa=0.55 to kappa=0.68. Likewise, intraobserver agreements increased from kappa=0.79 and 0.53 to kappa=0.81 and 0.70 for each reader, respectively. The areas under the ROC curves for each reader were 0.96 and 0.90, which were not significantly different. CONCLUSION: Including ABER view in routine sequences of MR arthrography increases the sensitivity, and inter- and intraobserver agreements for detecting partial thickness tear of rotator cuff tendon. PMID- 20001475 TI - An open-label, multicenter, phase 2a study to assess the feasibility of imaging metastases in late-stage cancer patients with the alpha v beta 3-selective angiogenesis imaging agent 99mTc-NC100692. AB - BACKGROUND: The alpha(v)beta(3) integrin is one of the angiogenesis-related membrane proteins highly expressed on the neovasculature of breast cancer and lung carcinomas. Labeling of the alpha(v)beta(3) integrin with (99m)Tc-NC100692 provides a potential tool for imaging angiogenesis and hence the presence of malignant lesions. PURPOSE: To determine the feasibility of detecting metastatic lesions in liver, lung, bone, and brain with scintigraphy using the alpha(v)beta(3)-avid imaging agent (99m)Tc-NC100692 in patients with breast or lung cancer, and to assess its safety profile. MATERIAL AND METHODS: Twenty-five patients--15 with lung cancer and 10 with breast cancer--were recruited at 10 centers. Metastases were newly diagnosed by computed tomography, magnetic resonance imaging, or bone scintigraphy, i.e., the reference standard. Patients underwent whole-body scans of approximately 10-15 min duration beginning at 45 min post-injection and a SPECT acquisition of approximately 30 min beginning at 75 min after injection of up to 1100 MBq (99m)Tc-NC100692. In case of liver metastases, dynamic acquisitions of 15 min were performed starting immediately post-injection. Safety measurements were performed up to 2.5 hours after injection and included hematology, serum biochemistry, coagulation, urine analysis, vital signs, oximetry, 12-lead ECG assessments, and a physical examination. RESULTS: In patients with breast cancer, (99m)Tc-NC100692 scintigraphy detected 1 of 7 liver, 4 of 5 lung, 8 of 17 bone, and 1 of 1 brain metastases. The corresponding numbers for lung cancer patients were 0 of 2, 17 of 18, 2 of 2, and 7 of 9. There was overall stability through the follow-up period for all investigated safety parameters. CONCLUSION: Scintigraphy with (99m)Tc NC100692 is feasible for detection of lung and brain metastases from breast and lung cancer, while the detection of liver and bone lesions is poor. The use of (99m)Tc-NC100692 is safe and well tolerated. PMID- 20001477 TI - The Henry Stonnington Award for best review paper published in Brain Injury 2008. PMID- 20001476 TI - Efficacy of plain radiography and computer tomography in localizing the site of pelvic arterial bleeding in trauma patients. AB - BACKGROUND: Immediate angiography is warranted in pelvic trauma patients with suspected arterial injury (AI) in order to stop ongoing bleeding. Prior to angiography, plain pelvic radiography (PPR) and abdominopelvic computer tomography (CT) are performed to identify fracture and hematoma sites. PURPOSE: To investigate if PPR and CT can identify the location of AI in trauma patients undergoing angiography. MATERIAL AND METHODS: 95 patients with pelvic fractures on PPR (29 women, 66 men), at a mean age of 44 (9-92) years, underwent pelvic angiography for suspected AI. Fifty-six of them underwent CT additionally. Right and left anterior and posterior fractures on PPR were registered, and fracture displacement was recorded for each quadrant. Arterial blush on CT was registered, and the size of the hematoma in each region was measured in cm(2). AIs were registered for anterior and posterior segments of both internal iliac arteries. Presence of fractures, arterial blush, and hematomas were correlated with AI. RESULTS: Presence of fracture in the corresponding skeletal segment on PPR showed sensitivity and specificity of 0.86 and 0.58 posteriorly, and 0.87 and 0.44 anteriorly. The area under the curve (AUC) was 0.77 and 0.69, respectively. Fracture displacement on PPR >0.9 cm posteriorly and >1.9 cm anteriorly revealed specificity of 0.84. Sensitivities of arterial blush and hematoma on CT were 0.38 and 0.82 posteriorly, and 0.24 and 0.82 anteriorly. The specificities were 0.96 and 0.58 posteriorly, and 0.79 and 0.53 anteriorly, respectively. For hematomas, the AUC was 0.79 posteriorly and 0.75 anteriorly. Size of hematoma >22 cm(2) posteriorly and >29 cm(2) anteriorly revealed specificity of 0.85 and 0.86, respectively. CONCLUSION: CT findings of arterial blush and hematoma predicted site of arterial bleeding on pelvic angiography. Also, PPR predicted the site of bleeding using location of fracture and size of displacement. In the hemodynamically unstable patient, PPR may contribute equally to effective assessment of injured arteries. PMID- 20001478 TI - A self-assessment questionnaire for mental fatigue and related symptoms after neurological disorders and injuries. AB - PRIMARY OBJECTIVE: The purpose was to evaluate a new self-reporting scale for assessment of mental fatigue and its usefulness in describing mental fatigue after neurological diseases and injuries. Mental fatigue is suggested to comprise a spectrum of items including sensory, emotional and cognitive symptoms. METHODS AND PROCEDURES: Mental fatigue and related symptoms were measured with a self assessment scale on different diagnostic groups with neurological injuries and diseases as well as healthy controls. The scale covers sleep, sensory, emotional and cognitive domains, mental recovery and 24-hour variation. RESULTS: There were significant correlations between all the questions in the self-assessing scale and also a high internal consistency among items (Cronbach's alpha of 0.944). Despite the different aetiologies of mental fatigue and related symptoms, the individuals rated the questions in a similar way. The total sum of scores, as well as the separate questions showed significantly lower values for healthy controls compared to mental fatigue groups. CONCLUSION: In spite of the different aetiology of mental fatigue, the items included in the scale were significant for different diseases with co-occurring mental fatigue. The self-reported scale is suggested to be a valuable tool for assessment of mental fatigue in different diagnoses. PMID- 20001479 TI - Exploring the study skills and accommodations used by college student survivors of traumatic brain injury. AB - PRIMARY OBJECTIVE: Survivors of severe traumatic brain injury (TBI) recovering sufficiently to attend post-secondary institutions often display learning skills differing from those of traditional students; because of this, many use formal and informal accommodations and learning strategies to support academic performance and social integration. Professionals know little about how accommodations and learning strategies affect educational experiences of college student survivors, how best to apply accommodations and learning strategies, how much survivors embrace them, the extent to which they facilitate college success or how they evolve over time. The purpose of this study was to explore these issues. RESEARCH DESIGN: The researchers collected and analysed qualitative data using a multiple case study design. METHODS AND PROCEDURES: Collected data included interviews, artifacts and field notes from four college student survivors, members of their families, college instructors and personnel from institution's offices for students with disabilities. MAIN OUTCOMES AND RESULTS: Findings suggest challenges, learning strategies, future perspectives and the need for academic accommodations fluctuate and evolve for many years post-injury. Findings also suggest some survivors perceive their learning capabilities more positively than do others associated with them, leading to questions about whether over-accommodation occurs and the potential negative ramifications associated with this practice. PMID- 20001480 TI - Traumatic brain injury and olfactory deficits: the tale of two smell tests! AB - PRIMARY OBJECTIVE: Olfactory functions are not systematically evaluated following traumatic brain injury (TBI). This study aimed at comparing two smell tests that are used in a clinical setting. RESEARCH DESIGN: The University of Pennsylvania Smell Identification Test (UPSIT) and the Alberta Smell Test were compared in terms of assessment time, cost and diagnosis. Parameters associated with olfactory loss such as injury severity, type of cerebral lesion and depressive data were considered. Forty-nine TBI patients admitted to an outpatient rehabilitation programme took part in this experiment. RESULTS: The scores of the two smell tests were significantly correlated. Both tests indicated that patients with frontal lesion performed significantly worse than patients with other types of lesion. Mood and injury severity were not associated with olfactory impairment when age was taken into account. Between 40-44% of the patients showing olfactory impairments were not aware of their deficit. CONCLUSIONS: Since a significant proportion of the patients showing olfactory impairments were not aware of their deficit, it is recommended than clinicians systematically evaluate olfactory functions using the Alberta Smell test. To refine their diagnosis, the UPSIT can also be used. PMID- 20001481 TI - Rehabilitation including treadmill therapy for patients with incomplete locked-in syndrome after stroke; a case series study of motor recovery. AB - PRIMARY OBJECTIVE: The aim of this study was to explore changes in motor function in patients with incomplete LIS referred to rehabilitation. RESEARCH DESIGN: A prospective, explorative, multiple case study design was applied. METHODS AND PROCEDURES: A multidisciplinary intervention programme was conducted including treadmill therapy (TT) with body weight support. Patients being able to stand upright supported by a standing frame were consecutively recruited to TT from 2001-2005. Physical performance was recorded on video and treadmill data were registered. Performances of transfer and walking were selected as the focus of observation. Two external observers described the performances independently, using an observational form and validated the joint and condensed descriptions. OUTCOMES AND RESULTS: A total of nine patients fulfilled the intervention programme and no adverse events were noted. Personal assistance and body weight support were reduced during the TT period and all patients demonstrated improved physical performance. While five patients were able to practice some kind of walking activity at the end of rehabilitation, four patients demonstrated improvement in body functions, showing better postural control. CONCLUSION: This study indicates that TT can be a safe and useful method to intensify the rehabilitation programme for patients with incomplete LIS. PMID- 20001482 TI - Isolated bilateral abducens nerves palsy due to traumatic delicate arteriovenous fistula. AB - BACKGROUND: Ocular motor dysfunction is common in patients with head trauma. Also, traumatic lateral gaze palsy is usually associated with brain stem lesion, peripheral nerve injury with or without basilar skull fracture and lateral rectus muscle injury or entrapment. However, isolated bilateral abducens nerve palsy is extremely rare. CASE STUDY: This study describes a case of isolated bilateral abducens nerve palsy in a 35-year-old male with head trauma without cervical and skull fractures. He was diagnosed with mild diffuse axonal injury. During his assessment, he developed mild limb weakness, an ataxic gait and impaired abductive movement in both eyes. Angiography of the left common carotid artery revealed multiple small arteriovenous fistulae in both inferior nasal conchae. After embolization, lateral gaze palsy of his eyes was significantly improved. CONCLUSIONS: Based on this case, arteriovenous fistula should be considered as a cause of bilateral isolated abducens nerve palsy. PMID- 20001483 TI - Mentoring programme for adolescent survivors of acquired brain injury. AB - PRIMARY OBJECTIVE: To report the findings of a mentor-adolescent relationship between two survivors of acquired brain injury (ABI). RESEARCH DESIGN: Case study report. METHODS AND PROCEDURES: The adolescent, a survivor of Eastern Equine Encephalitis, was paired with an adult mentor, a survivor of a TBI. Baseline scores on the Youth Quality of Life (YQOL), Wisconsin Quality of Life Index (WQLI) and the Mayo-Portland Adaptability Index-4 (MPAI-4) were recorded. EXPERIMENTAL INTERVENTIONS: The mentor provided support to the adolescent during the 10-week relationship conducted as a community-based programme for adults with acquired brain injury. In addition, both participants attended group activities that address the long-term needs of survivors of ABI. Post-programme scores were recorded on the YQOL, WQLI, MPAI-4 and a retrospective questionnaire. MAIN OUTCOMES AND RESULTS: The adolescent demonstrated improved quality of life on the YQOL and improved ability, adjustment and participation on the MPAI-4. The mentor demonstrated improved quality of life on the WQLI and improved adjustment and participation on the MPAI-4. Both participants indicated satisfaction with the programme on the retrospective questionnaire. CONCLUSIONS: The mentor programme provided enhanced quality of life and psycho-social support to both participants. The authors do caution, however, that these findings are preliminary and examination of the efficacy of such programming is ongoing. PMID- 20001484 TI - Pontobulbar palsy and sensorineural deafness (Brown-Vialetto-van Laere syndrome): the first case from Libya. AB - We report the first case of Brown-Vialetto-van Laere syndrome from Libya. A 27 year-old female patient with progressive pontobulbar palsy, sensorineural deafness and generalized muscle weakness is described. Although she was born to consanguineous first cousin parents, she was the only one affected among five brothers and six sisters. The first symptoms appeared at the age of 18 years with tinnitus, weakness and unsteadiness that progressed over the course of a few weeks to deafness, blurring of vision, dysphagia, and dysarthria. Over the last nine years her symptoms progressed slowly, although interrupted with periods of rapid deterioration. Fifty-eight cases have been reported since the first case was described in 1894. PMID- 20001485 TI - Voltage gated potassium channel antibodies in amyotrophic lateral sclerosis. AB - Although overlap exists between the clinical manifestations in patients with acquired peripheral nerve hyperexcitibility and amyotrophic lateral sclerosis (ALS), the presence of VGKC antibodies has not been associated with the development or clinical manifestations of motor neuron disease. We present the finding of elevated potassium channel antibodies in a cohort of patients with ALS and demonstrate a higher number of abnormal titer levels and higher mean titer levels in ALS patients compared to a cohort of patients with peripheral nervous system disorders. These results suggest the possibility of a subset of ALS patients where autoimmunity could play a role in disease development and progression. PMID- 20001486 TI - ALS with respiratory onset: clinical features and effects of non-invasive ventilation on the prognosis. AB - Respiratory muscle involvement is one of the main prognostic factors in amyotrophic lateral sclerosis (ALS). Acute respiratory failure is sometimes the first manifestation of the disease, although onset can be more insidious. In the present retrospective study, it was proposed to review the clinical features and to assess the effects of non-invasive ventilation (NIV) on the prognosis of patients with respiratory onset, which was taken to be present when the first symptoms of muscular weakness were dyspnoea at exertion, dyspnoea at rest, or orthopnoea. Respiratory onset ALS is uncommon, since it accounts for less than 3% of ALS cases. ALS with respiratory onset has some common clinical features: male predominance, frequent camptocormia or dropped head, frequent widespread fasciculations, limb mobility fairly well preserved and significant weight loss in the early stages. ALS patients with respiratory onset still have a poor prognosis compared with those with bulbar or spinal forms. NIV should be proposed promptly because it improves the symptoms, general state of health and survival time. Efforts should be made to inform general practitioners and chest physicians and remind them that neuromuscular respiratory insufficiency may be attributable to this particular form of ALS. PMID- 20001487 TI - Marital relationships in amyotrophic lateral sclerosis. AB - Our objective was to determine change over time in quality of marital relationships of people with ALS (pwALS). Fifty pwALS and their spouse carers completed measures of marital intimacy, pwALS's functional status, their own psychological status, and beliefs about the pwALS's illness. Twenty-seven dyads completed two further assessments, at six-monthly intervals. Post-ALS diagnosis ratings of current marital relationship did not change over time for pwALS or their spouses. However, spouse carers' ratings of their current marital relationship were lower than their ratings prior to illness onset. After adjusting for the pre-illness marital relationship, pwALSs' current ratings of their marital relationship were initially predicted by their self-rated psychosocial functional impairment and satisfaction with their personal relationships, and subsequently by levels of self-esteem. After adjusting for ratings of the pre-illness marital relationship, carers' current marital intimacy scores at first and second interview were predicted by ratings of the patients' psychosocial functional impairment and by feelings of burden at the third interview. In conclusion, quality of the pre-illness marital relationship is a significant predictor of ongoing marital relationship in both pwALS and their spouse carers. Additionally, social and psychological rather than disease symptoms are important predictors of marital relationship quality after ALS diagnosis. PMID- 20001488 TI - Pattern of spread and prognosis in lower limb-onset ALS. AB - Our objective was to establish the pattern of spread in lower limb-onset ALS (contra- versus ipsi-lateral) and its contribution to prognosis within a multivariate model. Pattern of spread was established in 109 sporadic ALS patients with lower limb-onset, prospectively recorded in Oxford and Sheffield tertiary clinics from 2001 to 2008. Survival analysis was by univariate Kaplan Meier log-rank and multivariate Cox proportional hazards. Variables studied were time to next limb progression, site of next progression, age at symptom onset, gender, diagnostic latency and use of riluzole. Initial progression was either to the contralateral leg (76%) or ipsilateral arm (24%). Factors independently affecting survival were time to next limb progression, age at symptom onset, and diagnostic latency. Time to progression as a prognostic factor was independent of initial direction of spread. In a regression analysis of the deceased, overall survival from symptom onset approximated to two years plus the time interval for initial spread. In conclusion, rate of progression in lower limb-onset ALS is not influenced by whether initial spread is to the contralateral limb or ipsilateral arm. The time interval to this initial spread is a powerful factor in predicting overall survival, and could be used to facilitate decision-making and effective care planning. PMID- 20001489 TI - Analysis of DPP6 and FGGY as candidate genes for amyotrophic lateral sclerosis. AB - DPP6 and FGGY genes have been recently associated with an increased susceptibility for sporadic amyotrophic lateral sclerosis. Here, we evaluated the role of these genes in ALS pathogenesis by undertaking a sequence analysis of a cohort of 190 ALS patients from France and Quebec. We did not observe any evidence that mutations in DPP6 and FGGY genes are involved in ALS. Our data indicate that mutations in these genes are unlikely to be a common cause of ALS in the French and French Canadian populations. PMID- 20001490 TI - Amyotrophic lateral sclerosis and sarcoidosis: a difficult differential diagnosis. PMID- 20001491 TI - Impairment of cardiac autonomic control in patients with amyotrophic lateral sclerosis. AB - The aim of this study was to investigate autonomic cardiac control in patients with amyotrophic lateral sclerosis (ALS). Fifty-five patients with sporadic ALS (28 female and 27 male; average age 56.00 +/- 10.34 years) were compared to 30 healthy controls (17 female and 13 male; average age 42.87 +/- 11.91 years). Patients with previous history of cardiac disease, diabetes mellitus, and impaired respiratory function were excluded from the study. Cardiovascular autonomic tests according to Ewing, power spectrum analysis of RR variability (low- and high-frequency bands - LF and HF, LF/HF index), real-time beat-to-beat ECG signal monitoring with heart rate variability analysis and baroreflex function analysis were carried out in all patients. Time-domain parameters of heart rate variability (mean RR interval, SDNN, SDANN, SDNN index, rMSSD and pNN50%) were obtained from 24-h ECG monitoring. ALS patients had a significantly higher score of sympathetic (p <0.01) and parasympathetic (p <0.001) dysfunction, as well as of the overall score of autonomic dysfunction (p <0.001). LF/HF index was significantly increased; baroreflex sensitivity and time-domain parameters of heart rate variability were highly significantly decreased in ALS patients (p <0.001). Our results demonstrated impaired cardiac autonomic control in ALS with marked parasympathetic dysfunction and sympathetic predominance. PMID- 20001492 TI - Practical guidance for dosing and switching paliperidone palmitate treatment in patients with schizophrenia. AB - OBJECTIVE: This article overviews the recommended dosing strategies for the treatment of schizophrenia patients using the recently FDA-approved once-monthly long-acting injectable atypical antipsychotic, paliperidone palmitate. METHODS: Using pharmacokinetic (PK), efficacy and safety data from clinical trials and a comprehensive population PK simulation model, dosing recommendations for paliperidone palmitate have been generated. RESULTS: The recommended initiation regimen is 150 mg eq. paliperidone palmitate (234 mg) on Day 1 followed by 100 mg eq. paliperidone palmitate (156 mg) on Day 8, each administered into the deltoid muscle, using a 1-inch 23 gauge (G) needle in those <90 kg and a 1.5-inch 22 G needle in those > or =90 kg. No oral supplementation is required. Monthly maintenance doses of paliperidone palmitate range from 25-150 mg eq. (39-234 mg; recommended dose of 75 mg eq. [117 mg]) injected into the deltoid (using weight adjusted needle) or gluteal (using 1.5 inch 22 G needle) muscle. The Day 8 dose may be administered +/-2 days and monthly doses +/-7 days, without a clinically significant impact on plasma concentrations. In patients with mild renal impairment (creatinine clearance [CrCL]: 50-80 mL/min), dosage should be adjusted. No dose adjustment is required in patients with mild or moderate hepatic impairment; no data currently exist regarding severe hepatic impairment. Elderly patients with normal renal function should receive the same dosage as younger adult patients with normal renal function. In the event of an age-related decline in CrCL, dosage should be adjusted accordingly. Paliperidone palmitate treatment can be initiated the day after discontinuing previous oral antipsychotic treatment. Paliperidone palmitate should be initiated at the next scheduled injection, and monthly thereafter, in patients switching from other long-acting injectable antipsychotics, including long-acting risperidone. CONCLUSIONS: These data provide practical guidance to clinicians on how to use paliperidone palmitate in adult patients with schizophrenia. PMID- 20001493 TI - Carcinoma of the nasal cavity and paranasal sinuses in Denmark 1995-2004. AB - OBJECTIVE: To evaluate the treatment outcome for sino-nasal carcinomas in Denmark from 1995-2004 and compare the results to the previous Danish survey covering 1982-1991. DESIGN: Retrospective follow-up. MATERIALS AND METHODS: In the five Danish head and neck oncology centres, charts of all consecutive patients with sino-nasal carcinomas were reviewed and data extracted to a common database. Altogether 242 patients from the period 1995-2004 were identified. Of these 162 (67%) were male and 80 (33%) female. Histologies included squamous cell carcinoma (55%), adenocarcinoma (28.5%), adenoid-cystic carcinoma (5.0%), undifferentiated carcinoma (4.5%), transitiocellular carcinoma (1.7%), mucoepidermoid carcinoma (0.8%), neuroendocrine carcinoma (2.5%), small cell carcinomas (1.2%) and carcinomas not otherwise specified (0.8%). Treatments included radiotherapy alone 79 (33%), surgery alone 29 (12%), combined surgery and radiotherapy 96 (40%), palliative/no treatment 38 (16%). A total of 204 (86%) patients were treated with curative intent. RESULTS: Of the 204 patients treated with curative intent, 94 (46%) relapsed. Most failures were in T-site (63, 30%). N-site failures were 10 (5%) and M-site failures six (3%). Failure occurring in T+N-site, T+M-site, N+M site and T+N+M-site were seven (3%), two (1%), one (0.5%) and five (3%) respectively. The 5-year actuarial local, nodal and loco-regional control rates were 55 + or - 4%, 86 + or - 3%, 49 + or - 4%, respectively. The overall 5-year actuarial survival rate for the entire cohort was 47 + or - 3%, and the corresponding cancer-specific 5-year actuarial survival rate was 57 + or - 3%. Female gender, nasal cavity tumour, adenocarcinoma and low clinical stage were significant positive prognostic factors in univariate analysis. A Cox multivariate analysis showed that only tumour site and clinical stage were independent significant prognostic factors. CONCLUSION: The current series has confirmed stage and tumour site as independent prognostic factors. Compared to the previous Danish survey covering the period 1982-1991, the overall survival and cancer-specific survival rates have improved significantly. PMID- 20001494 TI - The State of the Science: Informing choices across the cancer journey with public health mechanisms and decision processes. AB - Health decisions involve sharing information and making choices-even if the choice is to leave the decision to others. The way that information is delivered and understood by consumers across their cancer journey in turn influences the health decisions they take. A public health approach to the cancer journey considers the information needs of individuals and the structures and systems that facilitate the provision of credible and timely information. This paper examines emerging research that takes a public health approach to promote information-sharing and health decisions, identifies information-sharing mechanisms used by providers to facilitate shared decisions and evaluates decision support processes designed to improve information-sharing and self-care events. Evidence is presented to guide future research directions. PMID- 20001495 TI - Short time infusion of bevacizumab in combination with 5FU-based chemotherapy as first-line therapy in a non-selective patient group with metastatic colorectal cancer. PMID- 20001496 TI - Reversible stiff person syndrome presenting as an initial symptom in a patient with colon adenocarcinoma. PMID- 20001497 TI - Dosimetric comparison of postoperative whole pelvic radiotherapy for endometrial cancer using three-dimensional conformal radiotherapy, intensity-modulated radiotherapy, and helical tomotherapy. AB - BACKGROUND: The use of Intensity-modulated radiotherapy (IMRT) and Helical tomotherapy (HT) is increasing in gynecological cancer patients. No published studies have performed a dosimetric evaluation of whole pelvic radiotherapy (WPRT) using HT for postoperative endometrial cancer. The purpose of this study was to perform a direct dosimetric comparison of three-dimensional conformal radiotherapy (3D-CRT), IMRT and HT plans for WPRT in postoperative endometrial cancer patients, and to evaluate the integral dose to organs at risk (OARs) and normal tissue. MATERIAL AND METHODS: We selected ten patients with endometrial cancer undergoing postoperative WPRT. Plans for 3D-CRT, IMRT and HT were developed for each patient. All plans were normalized to deliver 50 Gy to 95% of the PTV. The dosimetry and integral dose to OARs and normal tissue were compared. The significance of differences was tested using a paired two-tailed Student t test. RESULTS: IMRT were superior to 3D-CRT in dose conformity (conformity index: 0.87 vs. 0.61, p = 0.00) and integral dose to OARs and normal tissue, although a greater volume of normal tissue receiving dose below 10 Gy was observed. The results were similar in HT except that the integral dose to normal tissue increased slightly. Compared directly with IMRT, HT showed better dose homogeneity and lower integral dose to rectum and bladder, but the integral dose to pelvic bones and normal tissue slightly increased. CONCLUSIONS: In postoperative WPRT of endometrial cancer, IMRT and HT result in better conformity and lower integral dose to OARs compared with 3D-CRT. The integral dose to normal tissue did not increase significantly in IMRT, although a greater volume of normal tissue is irradiated to the dose below 10 Gy. HT further improves the dose homogeneity and integral dose to rectum and bladder, at the expense of a slightly higher integral dose to pelvic bones and normal tissue. PMID- 20001498 TI - Carcinoid encephalopathy: A single entity or a spectrum of different disorders? PMID- 20001499 TI - SiRNA-mediated IGF-1R inhibition sensitizes human colon cancer SW480 cells to radiation. AB - PURPOSE: Insulin like growth factor receptor 1 (IGF-1R) is well-documented to play a key role in radiation response and tumor radiosensitivity, thus offering an attractive clinic drug target to enhance tumor sensitivity to anti-cancer radiotherapy. MATERIAL AND METHODS: Human colon carcinoma SW480 cells were transfected with the specific small interference RNA (siRNA) expression vector (pkD-shRNA-IGF-1R-V2) designed to target IGF-1R mRNA. The expression of IGF-1R mRNA and its protein among the transfected and untransfected cells were detected by semi-quantitative RT-PCR and ELISA assay. The changes in cell radiosensitivity were examined by MTT assay. RESULTS: Transfection of mammalian expression vector pkD containing IGF-1R siRNA was shown to reduce IGF-1R mRNA levels by up to 95%. ELISA assay detected a similar inhibition of IGF-1R protein levels in cells transfected with IGF-1R siRNA. SW480 cells transfected with the expression vector for siRNA significantly rendered cells more sensitive to radiation and the highest radiation enhancement ratio was 2.02 +/- 0.08. CONCLUSION: These data provide the first evidence that specific siRNA fragment (pkD-shRNA-IGF-1R-V2) targeting human IGF-1R mRNA is able to enhance colon cancer radiosensitivity. Also results indicated that, combining IGF-1R siRNA and radiation significantly enhances antitumor efficacy compared with either modality alone. PMID- 20001500 TI - Tumour differentiation grade is associated with TNM staging and the risk of node metastasis in colorectal cancer. AB - AIM: The tumour differentiation grade has been shown by numerous multivariate analyses to be a stage-independent prognostic factor in colorectal cancer. The aim of this study was to explore the importance of differentiation grading for the staging of colorectal cancer and how it relates to the components of the TNM system. MATERIAL AND METHODS: The study was a retrospective single-centre analysis of all patients undergoing surgical resection for colorectal cancer during the period 2002-2007 (n = 1239). The clinical parameters and pathology data of overall stage, differentiation grade, local tumour (T)-stage and metastasis status (M-stage) were included as well as the lymph node count of both assessed and metastatic nodes. The differentiation grade was correlated with demography, overall stage and each component of the TNM staging system. The correlation between differentiation grade and N-stage was also explored for the separate T-stages. RESULTS: The tumour differentiation grade correlated significantly with the overall TNM stage (p < 0.0001). The grade significantly correlated with the T-stage and the risk of having lymph node metastasis (p < 0.0001). A high grade was associated with a higher positive lymph node count in stage III disease (p < 0.0002). For the T-stages, the risk of node metastasis was significantly linked to the tumour grade. A low grade (G1) T2 had a 17% risk of lymph node metastasis compared to a 44% risk for a high grade (G4) T2. CONCLUSION: Tumour differentiation is an important prognostic factor. It correlates significantly with the overall stage of the TNM system and also to each of its components. The risk of having lymph node metastasis for each T-stage also correlates with the tumour grade. The findings can be of importance in postoperative risk assessment or when considering local resection procedures like TEM. PMID- 20001501 TI - A study of the expression of Cyclin E and its isoforms in tumor and adjacent mucosa, correlated to patient outcome in early colon cancer. AB - BACKGROUND: Cyclin E, a key regulator in the cell cycle, is often over-expressed in malignant disease. It can present as full length (FL) and low-molecular-weight (LMW) isoforms. The purpose of this study was to characterize the expression pattern of cyclin E in colon cancer, both in tumor and in macroscopically normal adjacent mucosa. A secondary aim was to study the possible correlation to clinical factors and patient outcome. MATERIAL AND METHOD: Tumor and mucosa tissue from 114 patients with radically operated, non-metastatic colon tumors were analyzed. The cyclin E expression was measured by Western Blot in the tumor and adjacent mucosa using the antibody targeting C-terminal. The cyclin E expression was correlated to both pathology factors as differentiation grade and to the patient outcome. RESULTS: Cyclin E was detected in both tumor and adjacent mucosa and in both FL and LMW-forms. FL was present in 29 (25.4%) tumors and only in three (2.6%) mucosa samples, the corresponding figures for the LMW-isoforms were 80 (70.2%) and 67 (58.8%). There was no correlation between the cyclin E expression and gender, age, tumor location or tumor pathology. Patients with a high expression of LMW isoforms (p < 0.03) or a high total expression (FL+LMW) (p < 0.006) had higher risks of recurrence and thus a worse survival. CONCLUSION: Cyclin E is expressed in FL- and LMW-forms in both colon tumors and the macroscopically normal adjacent mucosa. A high expression of cyclin E in tumor was associated with an increased risk of tumor recurrence and a worse outcome. It could be a possible prognostic marker in non-metastatic colon cancer. PMID- 20001502 TI - Inhibition of enhanced green fluorescent protein expression by (dextran hexamethylenediisocyanate)-g-polyethylenimine/siRNA complexes. AB - Small interfering RNA (siRNA), as a potential tool for gene therapy, requires suitable vehicles for stable complexation, protection, low cytotoxicity and high efficiency of gene knockdown. Here, a new polycation, (dextran hexamethylenediisocyanate)-g-polyethylenimine ((Dex-HMDI)-g-PEI), was used as a siRNA vector to knock down the expression of Enhanced Green Fluorescent Protein (EGFP). It was found that the complexation of siRNA with the polycation can significantly knock down the expression of EGFP, but has no remarkable effect on the expression of luciferase. PMID- 20001503 TI - Enhanced stability and in vitro bioactivity of surfactant-loaded liposomes containing Asiatic Pennywort extract. AB - The objective of this work has been the microencapsulation of Asiatic Pennywort (AP) extract with lecithin from soybean. The effect of various quantities of non ionic surfactant (Montanov82) on liposomes upon physicochemical characteristics as well as their in vitro bio-activities was investigated. An addition of surfactant resulted in a decrease in particle size and an increase in percentage AP entrapment efficiency of liposomes. The surfactant-loaded liposomes demonstrated higher stability than surfactant-free liposomes where higher percentage AP remaining of liposomes can be achieved depending on surfactant concentration. No significant difference was found on AP release profiles among varied surfactant concentrations, although a presence of surfactant resulted in prolonged AP release rate. Liposomal AP with 20% w/w surfactant or higher demonstrated low cytotoxicity, a stronger anti-oxidation effect and collagen production on dermal fibroblast cells when compared with free AP and surfactant free liposomes, possibly due to better cell internalization and less AP degradation in cells. PMID- 20001504 TI - Socioeconomic differences in incidence of cervical cancer in Finland by cell type. AB - INTRODUCTION: We studied variation in incidence of cervix cancer during 1971-1995 among Finnish women born in 1906 to 1945 by social class and cell type. MATERIAL AND METHODS: The Finnish Cancer Registry data were linked to the 1970 Population Census, which included social class data. There were 0.8 million individuals in the cohort under follow-up, with 5,102 squamous cell cancers and 573 cases of cervical adenocarcinoma diagnosed after the census date. RESULTS: Incidence of squamous cell cancer was more than two-fold in the lowest social class (standardized incidence ratio (SIR) 1.29, 95% CI 1.21-1.36) as compared with the highest one (SIR 0.59, 0.51-0.66), while there was no association between social class and risk on adenocarcinoma (SIR 1.07, 0.87-1.28 and 1.08, 0.79-1.45, respectively). DISCUSSION: Oncogenic HPV is regarded as the necessary cause of all types of cervix cancer. Sexually transmitted diseases and social status are correlated. Lack of association between adenocarcinoma and social class makes the HPV-etiology of this cell type less credible than that of squamous cell cancer. PMID- 20001505 TI - Galectin-3 detection on large-needle aspiration biopsy improves preoperative selection of thyroid nodules: a prospective cohort study. AB - BACKGROUND: New techniques of improving diagnostic reliability of thyroid nodules are needed. AIM AND METHODS: This prospective cohort study includes patients with one (201) or multiple (22) palpable nodule(s). Preoperative fine-needle aspiration biopsy (FNAB), large-needle aspiration biopsy (LNAB), and galectin-3 detection on LNAB (GAL-3-LNAB) (total of 245 nodules) were compared when the FNAB finding was 'inadequate' or 'indeterminate'. The sizes of the needles used for FNAB and LNAB were compared with the size of thyroid follicles. Forty nodules were surgically excised according to current recommendation. RESULTS: GAL-3-LNAB was inadequate in 4% of nodules, compared with 34% using FNAB and 11% using LNAB (P < 0.0001). GAL-3-LNAB showed no indeterminate findings, compared with 15% using FNAB and 13% using LNAB (P < 0.0001). Among the 40 excised nodules, GAL-3 LNAB showed the highest accuracy values. The sensitivity (P = 0.011) and specificity (P < 0.000; P = 0.001) ranges were 40%-100% and 20%-40% for FNAB, 40% 100% and 50%-53.7% for LNAB, and 100% and 76.7%-80% for GAL-3-LNAB, respectively. The largest needles used for LNAB, 20 or 18 gauge, with an internal diameter of 0.6 or 0.91 mm, recorded the lowest rate of inadequate or indeterminate FNAB findings. CONCLUSIONS: GAL-3-LNAB reduced inadequate, abolished indeterminate findings, and provided specificity values higher than FNAB or LNAB in palpable thyroid nodules. PMID- 20001506 TI - Pattern recognition of vertical strabismus using an artificial neural network (StrabNet). AB - BACKGROUND: Our goal is to develop an accessible expert system (StrabNet) that will assist in the clinical diagnosis of vertical strabismus, form the basis of a teaching/learning tool, and contribute to the audit process. Potentially, this model can be extended to other strabismus deviations. METHODS: Vertical deviations were separated into eight classifications (diagnoses). An expert system based on architecture of artificial neural networks learned the patterns for each class of deviation based on 10 prism cover-test measurements (9 cardinal positions and near fixation). The expert system was tested with previously unseen and real-patient data. This system was extended to a reduced model requiring only six measurements (primary position, right, left, up, down gaze, and near fixation), and evaluated with real patient data. A freely available Web implementation is available on the Internet at www.StrabNet.com . RESULTS: The expert system was found to be highly accurate at diagnosing vertical strabismus (100% and approximately 96% for ten and six measurement models, respectively) from one of the eight classes. CONCLUSION: StrabNet is of demonstrable value in diagnosing commonly presenting vertical deviations from prism cover test (PCT) measurements. Its potential role in teaching and in audit is identified. PMID- 20001507 TI - Complications of botulinum toxin a and their adverse effects. AB - AIM: To evaluate the prevalence of complications after low doses (< 3 units) of botulinum toxin (BT) injection to horizontal extraocular muscles for diagnostic and therapeutic purposes. METHODS: A case record review was undertaken of patients treated with BT (Dysport) since 2000. Data were extracted for gender, age, diagnosis, number of BT injections, dose, recorded complications, and time to recovery. RESULTS: Two hundred eleven (211) cases were reviewed, 48.5% children, 51.5% adults (45.5% male, 54.5% female). Diagnoses included constant and intermittent esotropia and exotropia, decompensating esophoria, exophoria and microtropia, accommodative spasm, mechanical restrictions, and cranial nerve palsy. One hundred sixty-three (163) patients had one injection; 38 had two; 7 had three; 2 had four; 1 had six; for a total of 274 injections. BT dose was 1 unit in four cases; 2.5 units in two; 3 units in seven; and 2 units in the remainder. At 2 weeks follow-up, six cases had induced vertical deviation; three had subconjunctival haemorrhage. At 2 weeks, 23 cases had ptosis, but a further 10 cases had ptosis that had resolved by 2 weeks follow-up. All complications had resolved by 6 weeks follow-up. CONCLUSIONS: A low incidence of complications per injection (12.4%) was found particularly for induced vertical deviations (2%) and hemorrhages (1%). Incidence for ptosis was the highest (8.4%). Notably, 10 additional cases of ptosis had resolved before follow-up. Two children were given occlusion therapy to prevent the possibility of stimulus deprivation occlusion until ptosis resolution. PMID- 20001508 TI - Vergence position of rest: spontaneous variability. AB - BACKGROUND: We determined the vergence position of rest (horizontal component) in two sessions, 1 to 6 weeks apart. Each session contained 8 trials. Twenty observers with normal eyes looked through Risley prisms that allowed a continuous modulation of their strength. We asked the observers to look at a fully fusionable picture, and to adjust the prisms such that viewing appeared most relaxing. The viewing distances were 400 cm and 50 cm. RESULTS: The standard deviation among the 8 trials of each session, averaged over the two sessions, the viewing distances of 400 cm and 50 cm, and the 20 observers, was + or -1.4 prism dioptres (cm/m). From the first session to the second, many observers changed the preferred prism significantly. At a viewing distance of 400 cm, 10 of 20 observers changed their preferred prism by more than 1.0 prism diopter, up to 3.3 prism diopters. At a viewing distance of 50 cm, 11 of 20 observers changed their preferred prism by more than 1.0 prism diopter, up to 5.1 prism diopters. CONCLUSIONS: The prismatic strength estimated as being most relaxing can be very variable. Before considering a prescription, the practitioner should determine whether the preferred prismatic strength remains reasonably stable. To ascertain this consistency, more than one session seems to be necessary. PMID- 20001509 TI - Visual outcome in 879 children treated for strabismus: insufficient accommodation and vision deprivation, deficient emmetropisation and anisometropia. AB - PURPOSE: To identify the effect of infant hypermetropia on residual amblyopia in children remaining after treatment. MATERIALS AND METHODS: 879 strabismic children had cycloplegic retinoscopy at the age of 6 months and later when strabismus was diagnosed. A total of 26 hypermetropes consistently wore glasses from the age of 6 months, and 305 other hypermetropes had their accommodation periodically assessed by dynamic retinoscopy before strabismus was diagnosed. The relation between the last known visual acuity after treatment and all other clinical findings was analysed using t-tests and regression analysis. RESULTS: Vision less than 6/12 in nonfixing eyes was associated with infantile hypermetropia > +5.0D, and in rare instances in excess of -3.0D of myopia. In hypermetropes only, anisometropia was associated with worse amblyopia. Astigmatism was associated with myopia and low levels of hypermetropia but not with worse vision. The vision of fixing eyes that were hypermetropic was significantly worse than that of emmetropic or myopic eyes. Wearing spectacles from the age of 6 months was associated with better vision in the nonfixing eye. Vision in the nonfixing eye of 19 hypermetropic heterotropes who started wearing glasses between 6 and 18 months of age also was better than that of those who started wearing glasses after 18 months of age. Insufficiency of accommodation was related to the degree of infantile hypermetropia, to worse vision and to failure to emmetropise. Difference in amplitude of accommodation between the eyes was largest in hypermetropes with anisometropia > +1.50 D and was marginally associated with worse vision in microtropes who became anisometropic after infancy. Hypermetropia decreased more in fixing eyes than in nonfixing eyes, thereby causing anisohypermetropia to increase after infancy. This change was most pronounced in hypermetropic microtropes. Conversely, failure of fixing eyes to emmetropise was associated with an increase in their inability to accommodate and heterotropia. CONCLUSIONS: On the basis of these findings, severe residual amblyopia in children remaining after treatment could be explained by additional vision deprivation. It can be reduced by starting spectacle correction of hypermetropia before the age of 18 months. Anisometropia seemed the result of deficient emmetropisation. PMID- 20001510 TI - Nightblindness-associated transient tonic downgaze (NATTD) in infant boys with chin-up head posture. AB - Eleven infant boys presented with chin-up head posture, tonic downgaze and, on attempted upgaze, large-amplitude upward saccades with deceleration during the slow phase downward. The gaze-evoked upward saccades disappeared at the age of 2 or 3 years. In addition, they had high-frequency, small-amplitude horizontal pendular nystagmus that remained. Among these infant boys were 2 pairs of maternally related half-brothers, 2 cousins, and 2 siblings. Visual acuity ranged from 0.1 to 0.6, ERG-amplitudes (both A- and B-wave) were reduced, and severe myopia was found in 5 cases. Eight boys had CACNA1F mutations, and 1 boy had a NYX mutation, compatible with incomplete or complete congenital stationary nightblindness (iCSNB or cCSNB), respectively. This points to a defective synapse between the rod and the ON-bipolar cell causing the motility disorder: CACNA1F is located on the rod side of this synapse, whereas NYX is located on the side of the ON-bipolar cell. The coexistence of horizontal and vertical nystagmus has been previously described in dark-reared cats. PMID- 20001511 TI - Beware angle kappa: an unusual problem following strabismus surgery. AB - We report the case of a 33-year-old male with a negative angle kappa which became apparent following surgery for a consecutive exotropia. PMID- 20001512 TI - Opaque intraocular lens for intractable diplopia-UK survey. AB - AIMS: To assess the practice of the use of an opaque intraocular lens (IOL) for intractable diplopia in the UK. METHODS: A questionnaire was sent to 892 consultant ophthalmologists in practice requesting information on the treatment of intractable diplopia with an opaque IOL. Respondents were asked whether they would consider using an opaque IOL for intractable diplopia. Information was gathered on the numbers of lenses implanted, the type of lens used, and whether the lens was implanted inside or outside the capsular bag. In addition, we asked for the causes of intractable diplopia, the success in eliminating diplopia, the use of postoperative pilocarpine, and whether postoperative surveillance was part of the follow-up protocol. RESULTS: Of the 481 completed questionnaires received, 72% would consider implanting an opaque IOL. A total of 48 surgeons had implanted 1 or more lenses. There were 97 patients who were implanted with an opaque IOL over the past 15 years. Strabismus, nerve palsies, and previous retinal detachment surgery with diplopia were the main indications. Pseudophakic lenses were used by 38 surgeons in the capsular bag, 6 used iris-supported phakic lenses, and 2 used phakic lenses in the anterior chamber angle. The use of postoperative pilocarpine was reported by 9 surgeons, and 6 surgeons used regular ultrasound fundal surveillance. Patients were reported to be completely asymptomatic at discharge by 31 surgeons, with 15 surgeons reporting patients who were still symptomatic postoperatively. Only 1 intraoperative complication was reported. CONCLUSIONS: An opaque IOL is a safe method for treating intractable diplopia. This survey confirms that its practice is widely accepted in the UK. PMID- 20001513 TI - Alternate day exotropia. AB - INTRODUCTION: Cyclic strabismus is a rare disorder in which manifest strabismus and controlled latent deviation (heterophoria) alternates on a regular pattern which is usually over a 48-hr period. During the "straight" period the patient has a fully controlled heterophoria for all distances, with very good fusion and stereopsis. During the "squinting" period the patient has a constant manifest strabismus for all distances, with no demonstrable fusion and stereopsis. METHOD: We describe 2 cases of alternate day exotropia. One patient requires strabismus surgery to obtain constant binocularity. The second patient underwent surgery for an intermittent exotropia and subsequently developed an alternate day exotropia. The condition very gradually reduced over the years and the patient is now fully binocular. CONCLUSION: We report the first 2 cases of cyclic exotropia in non Japanese female children presenting from about 1 to 3 years of age, without ocular or systemic pathology. PMID- 20001514 TI - Experiences with the elimination of amblyopia and the restoration of binocular vision in squinters, part III. PMID- 20001515 TI - Temporal reduction of HIV type 1 viral load in breast milk by single-dose nevirapine during prevention of MTCT. AB - Short-course zidovudine (ZDV) with or without a single dose of nevirapine (sdNVP) is widely used to prevent mother-to-child HIV transmission (PMTCT). However, more data on viral load in breast milk following pMTCT regimens are needed. In a randomized PMTCT study in Botswana, in which half of the women received sdNVP in labor, stored samples from mothers assigned to breastfeed were analyzed for HIV-1 RNA in breast milk supernatant. A total of 527 samples from 282 women, collected at delivery, 2 weeks, 2 months, and 5 months postpartum were available for testing. Cell-free breast milk HIV-1 RNA was detectable (>40 copies/ml) in 44.8% (236/527) of samples analyzed. Women randomized to sdNVP + ZDV were more likely to have undetectable breast milk viral loads at 2 weeks postpartum compared with those who received ZDV alone (67.8% vs. 38.5%, p = 0.002). By 2 months postpartum the difference between study arms disappeared, and 43.8% of women who received sdNVP + ZDV had undetectable HIV-1 RNA compared to 53.8% of the ZDV alone group (p = 0.19) and 60.5% vs. 64.5%, respectively, at month 5 (p = 0.61.) The addition of sdNVP to antenatal short-course AZT resulted in significantly reduced breast milk viral loads at 2 weeks postpartum suggesting a reduced risk of MTCT during the early postpartum period. However, viral loads in both study arms were comparable at 2 and 5 months postpartum, suggesting that the receipt of sdNVP in labor may defer rather than blunt the postpartum viral load rebound seen in breast milk after the discontinuation of ZDV. PMID- 20001516 TI - Adiponectin and leptin levels in Chinese patients with HIV-related lipodystrophy: a 30-month prospective study. AB - The relationship of adipocytokine with the development of HIV-related lipodystrophy was investigated in a case-control study. Adipocytokine, lipid, and glycemic parameters were measured at every visit. Logistic regression analysis was used to assess the HIV-LD risk factors and the Spearman correlation coefficients test was used to assess the correlation between adiponectin with other metabolic variables. Most of the patients (96.3%) developed HIV-LD after month 12. Comparing the baseline adiponectin, the adiponectin concentration of the HIV-LD group rose by month 6 and began to decrease substantially by month 18; this reduction was maintained until month 30 (p < 0.05). Comparing the HIV-NLD group, the adiponectin concentration at months 18, 24, and 30 were significantly lower in the HIV-LD group. The leptin concentration of both the HIV-LD and HIV NLD groups remained stable. Patients in the lower concentration of baseline adiponectin and greater adiponectin change rate at month 18 presented with increased odds ratio for HIV-LD. The adiponectin level had a correlation with serum triglycerides (r = -0.616, p < 0.0001), serum insulin concentration (r = 0.494, p = 0.001), and HDL-C (r = 0.673, p < 0.0001). The adiponectin concentration of HIV-LD began to decrease substantially by month 18. The lower baseline concentration of adiponectin and the greater change rate at month 18 were independent risk factors of HIV-LD. The adiponectin level had a correlation with serum triglycerides, serum insulin concentration, and HDL-C, suggesting that adiponectin may link the metabolic abnormalities and HIV-LD. PMID- 20001517 TI - Short communication: transmission of hepatitis B viruses with lamivudine resistance mutations in newly diagnosed HIV individuals. AB - From 1519 newly diagnosed HIV individuals seen in Madrid between the years 2000 and 2008, 65 (4.3%) were HBsAg(+). Two HIV/HBV-coinfected patients showed the lamivudine resistance mutation M204V in HBV while no drug resistance mutations were recognized in HIV. None of them admitted prior exposure to antiretroviral drugs. Thus, HIV/HBV-coinfected patients might benefit from baseline drug resistance testing for both HIV and HBV to optimize the selection of anti-HBV active antiviral therapy. PMID- 20001519 TI - Short communication: oxidative stress in HIV-infected individuals: a cross sectional study. AB - HIV infection increases the oxidative stress process, and antiretroviral combination therapy increases protein oxidation and preexistent oxidative stress. The latter induces production of reactive oxygen species. Lipid peroxidation (LPO) is a means of determining oxidative stress. There is also a deficiency of glutathione in HIV infection. Persistent oxidative load leads to an accelerated rate of consumption of glutathione (GSH). This study measured LPO and GSH levels in plasma of HIV-infected individuals with or without therapy and compared these with healthy controls. One hundred HIV-infected individuals and 30 healthy controls were included in the study. LPO and GSH levels were measured in plasma according to previously described methods. The mean level of LPO in HIV-infected individuals was 0.7 +/- 0.1 micromol/ml (range, 0.5-0.9 micromol/ml), whereas the mean LPO level in controls was 0.3 +/- 0.1 micromol/ml (range, 0.2-0.4 micromol/ml). The mean LPO levels were significantly higher in HIV-infected individuals as compared to healthy controls (p value <0.0001). The mean GSH level in HIV-infected individuals was 0.06 +/- 0.01 micromol/ml (range, 0.03-0.08). The mean GSH level in healthy controls was 0.09 +/- 0.01 micromol/ml (range, 0.05 0.1). The mean glutathione level in HIV-infected individuals was significantly lower in compared to healthy controls (p value < 0.0001). There was a significant positive correlation between absolute CD4 cells and GSH levels (rho = 0.182, p = 0.045). There is increased oxidative stress in HIV-infected patients. Whether supplementation with antioxidants will reduce this oxidative stress is still unknown. PMID- 20001518 TI - Early versus delayed fixed dose combination abacavir/lamivudine/zidovudine in patients with HIV and tuberculosis in Tanzania. AB - Fixed dose combination abacavir/lamivudine/zidovudine (ABC/3TC/ZDV) among HIV-1 and tuberculosis (TB)-coinfected patients was evaluated and outcomes between early vs. delayed initiation were compared. In a randomized, pilot study conducted in the Kilimanjaro Region of Tanzania, HIV-infected inpatients with smear-positive TB and total lymphocyte count <1200/mm(3) were randomized to initiate ABC/3TC/ZDV either 2 (early) or 8 (delayed) weeks after commencing antituberculosis therapy and were followed for 104 weeks. Of 94 patients screened, 70 enrolled (41% female, median CD4 count 103 cells/mm(3)), and 33 in each group completed 104 weeks. Two deaths and 12 serious adverse events (SAEs) were observed in the early arm vs. one death, one clinical failure, and seven SAEs in the delayed arm (p = 0.6012 for time to first grade 3/4 event, SAE, or death). CD4 cell increases were +331 and +328 cells/mm(3), respectively. TB immune reconstitution inflammatory syndromes (TB-IRIS) were not observed in any subject. Using intent-to-treat (ITT), missing = failure analyses, 74% (26/35) vs. 89% (31/35) randomized to early vs. delayed therapy had HIV RNA levels <400 copies/ml at 104 weeks (p = 0.2182) and 66% (23/35) vs. 74% (26/35), respectively, had HIV RNA levels <50 copies/ml (p = 0.6026). In an analysis in which switches from ABC/3TC/ZDV = failure, those receiving early therapy were less likely to be suppressed to <400 copies/ml [60% (21/35) vs. 86% (30/35), p = 0.030]. TB-IRIS was not observed among the 70 coinfected subjects beginning antiretroviral treatment. ABC/3TC/ZDV was well tolerated and resulted in steady immunologic improvement. Rates of virologic suppression were similar between early and delayed treatment strategies with triple nucleoside regimens when substitutions were allowed. PMID- 20001521 TI - CRF45_AKU, a circulating recombinant from Central Africa, is probably the common ancestor of HIV type 1 MAL and HIV type 1 NOGIL. AB - Abstract In this study, we characterized four HIV-1 strains from Cameroon, Gabon, and the Democratic Republic of Congo (DRC), collected during independent serosurveys, and previously found to cluster in the pol gene with HIV-1 MAL and HIV-1 NOGIL3, two complex recombinant viruses reported in the early HIV epidemic, and with the recombinant strain 04FR.AUK recently described in France. The four newly sequenced viruses shared the same structure as 04FR.AUK, involving alternating fragments of subtype A, K, and unclassified (U) fragments, representing a new CRF called CRF45_AKU. Some of the unclassified fragments were related to unclassified regions described in either CRF04 or CRF09 strains. Careful reanalysis of HIV-1 MAL and HIV-1 NOGIL3 demonstrated that these strains were related exclusively to CRF45_AKU and either two subtype D fragments for HIV 1 MAL or one subtype H segment for HIV-1 NOGIL3. Following extensive blast searches, related gag, pol, and env sequences were observed in Central and West Africa (Senegal, Mali), as well as in Europe (France, Spain, Italy, Cyprus), Argentina, and China. PMID- 20001522 TI - Diffuse centripetal vesicular rash. Kaposi varicelliform eruption (KVE). PMID- 20001520 TI - Systemic dendritic cell mobilization associated with administration of FLT3 ligand to SIV- and SHIV-infected macaques. AB - Reports indicate that myeloid and plasmacytoid dendritic cells (mDCs and pDCs), which are key effector cells in host innate immune responses, can be infected with HIV-1 and are reduced in number and function during the chronic phase of HIV disease. Furthermore, it was recently demonstrated that a sustained loss of mDCs and pDCs occurs in SIV-infected macaques. Since loss of functional DC populations might impair innate immune responses to opportunistic microorganisms and neoplastic cells, we explored whether inoculation of naive and SIV- or SHIV infected pigtailed macaques with the hematopoietic cytokine FLT3-ligand (FLT3-L) would expand the number of mDCs and pDCs in vivo. After the macaques received supraphysiologic doses of FLT3-L, mDCs, pDCs, and monocytes increased up to 45 fold in blood, lymph nodes, and bone marrow (BM), with DC expansion in the BM preceding mobilization in blood and lymphoid tissues. FLT3-L also increased serum levels of IL-12, at least transiently, and elicited higher surface expression of HLA-DR and the activation markers CD25 and CD69 on NK and T cells. During and after treatment of infected animals, APCs increased in number and were activated; however, CD4(+) T cell numbers, virion RNA, and anti-SIV/SHIV antibody titers remained relatively stable, suggesting that FLT3-L might be a safe modality to expand DC populations and provide therapeutic benefit during chronic lentivirus infections. PMID- 20001524 TI - High serum cholesterol levels are associated with herpes zoster infection after heart transplantation. PMID- 20001525 TI - Histoplasmosis in solid organ transplant recipients. PMID- 20001527 TI - Reducing the burden of tuberculosis presenting during the initial months of antiretroviral therapy in resource-limited settings. PMID- 20001529 TI - Presence of high-titer neutralizing antibodies against enterovirus 71 in intravenous immunoglobulin manufactured from Chinese donors. PMID- 20001530 TI - First report of hypervirulent strains polymerase chain reaction ribotypes 027 and 078 causing severe Clostridium difficile infection in Italy. PMID- 20001531 TI - Idiopathic noncirrhotic portal hypertension in HIV-infected patients. PMID- 20001535 TI - In vivo selection of biocompatible alginates for islet encapsulation and subcutaneous transplantation. AB - Islet encapsulation requires several properties including (1) biocompatibility, (2) immunoprotection, and (3) oxygen diffusion for islet survival and diabetes correction. New chemical alginates were tested in vivo and compared with traditional high-mannuronate and -guluronate alginates. New alginates with coupled peptide sequence (sterile lyophilized high mannuronate [SLM]-RGD3% and sterile lyophilized high guluronate [SLG]-RGD3%), to improve encapsulated cell adherence in the matrix, and alginates with a very low viscosity (VLDM7% and VLDG7%), to reduce implant size by loading a higher number of islets per volume of polymer, were implanted subcutaneously in 70 Wistar rats for comparison with alginates of high viscosity and high content of mannuronic (SLM3%) or guluronic acids (SLG3%). Permeability of alginates to 36-, 75-, and 150-kDa lectins coupled to fluorescein isothiocynate was quantified before implantation and at 2, 4, and 12 weeks after implantation. Biocompatibility (fibrosis, graft stability, immunologic infiltration by CD3/CD68 cells, and neovascularization) was assessed at each explantation time. Permeability to small molecules was found for all alginates. Impermeability to 150-kDa molecules, such as IgG, was observed only for SLM3% before implantation and was maintained up to 12 weeks after implantation. SLM3% and SLG3% demonstrated better graft stability with lower CD3/CD68 recruitment and fibrosis than the other alginates. SLM3% induced a significantly higher angiogenesis and maintained oxygen pressure at approximately 40 mm Hg for up to 4 weeks after implantation as measured by in vivo electronic paramagnetic resonance oximetry. SLM-encapsulated pig islets implanted subcutaneously in rats demonstrated no inflammatory/immunologic reactions and islets functioned for up to 60 days without immunosuppression. A traditional alginate made of high mannuronic content (SLM3%) is an adapted material to immunoprotect islets in subcutaneous tissue. No improvement was found with lower viscosity and use of GRGDSP-peptide sequence. PMID- 20001536 TI - Electro-acupuncture for pain relief after nasal septoplasty: a randomized controlled study. AB - BACKGROUND: Electro-acupuncture (EA) can be effective in some pain conditions, but still there have been no randomized controlled studies of EA for treatment of acute postoperative pain in nasal septoplasty. Therefore, we did a study to test whether EA of specific points is superior to sham acupuncture for complementary analgesia after nasal septoplasty. METHODS: Ninety (90) patients with an American Society of Anesthesiology (ASA) physical status of I-II scheduled for elective septoplasty were randomly allocated to acupuncture group or control group. Postoperative pain was treated with intravenous meperidine. The time to the first analgesic request, and pain intensity on a 100-mm visual analogue scale (VAS-100) were used to evaluate postoperative pain, and the amount of postoperative meperidine and incidence of analgesia related to side-effects were recorded. RESULTS: Postoperative pain intensity on VAS-100 and respiratory depression were similar in both groups (p > 0.05), but nausea and vomiting were less in the acupuncture group than in the control group (p < 0.05). CONCLUSIONS: Findings from our study demonstrate that both uses of EA and 0.1 mg/kg IV morphine given intraoperatively resulted in a similar postoperative pain score, and meperidine was not given in any patient in the two groups during postoperative period because the VAS scores of all patients were less than 40 mm. PMID- 20001537 TI - Efficacy and limitation of a chlorhexidine-based decolonization strategy in preventing transmission of methicillin-resistant Staphylococcus aureus in an intensive care unit. AB - BACKGROUND: Surface-active antiseptics, such as chlorhexidine, are increasingly being used as part of intervention programs to prevent methicillin-resistant Staphylococcus aureus (MRSA) transmission, despite limited evidence and potential for resistance. We report on the effect of an antiseptic protocol on acquisition of both endemic MRSA and an outbreak strain of MRSA sequence type 239 (designated TW). METHODS: Interrupted time-series data on MRSA acquisitions in two 15-bed intensive care units were analyzed using segmented regression models to estimate the effects of sequential introduction of an educational campaign, cohorting, and a chlorhexidine-based antiseptic protocol on transmission of TW and non-TW MRSA strains. Representative TW and non-TW MRSA strains were assessed for carriage of qacA/B genes and antiseptic susceptibility. RESULTS: The antiseptic protocol was associated with a highly significant, immediate 70% reduction in acquisition of non-TW MRSA strains (estimated model-averaged incidence rate ratio, 0.3; 95% confidence interval, 0.19-0.47) and an increase in acquisition of TW MRSA strains (estimated model-averaged incidence rate ratio, 3.85; 95% confidence interval, 0.80-18.59). There was only weak evidence of an effect of other interventions on MRSA transmission. All TW MRSA strains (21 of 21 isolates) and <5% (1 of 21 isolates) of non-TW MRSA strains tested carried the chlorhexidine resistance loci qacA/B. In vitro chlorhexidine minimum bactericidal concentrations of TW strains were 3-fold higher than those of non-TW MRSA strains, and in vivo, only patients with non-TW MRSA demonstrated a reduction in the number of colonization sites in response to chlorhexidine treatment. CONCLUSION: A chlorhexidine-based surface antiseptic protocol can interrupt transmission of MRSA in the intensive care unit, but strains carrying qacA/B genes may be unaffected or potentially spread more rapidly. PMID- 20001538 TI - Prevention and control of methicillin-resistant Staphylococcus aureus: dealing with reality, resistance, and resistance to reality. PMID- 20001539 TI - Treatment of HIV-related inflammatory cerebral cryptococcoma with adalimumab. AB - Cryptococcomas have been described in AIDS patients in the setting of immune reconstitution inflammatory syndrome. We report the first case of human immunodeficiency virus-related inflammatory cerebral cryptococcoma to be treated with a recombinant human monoclonal tumor necrosis factor antagonist. PMID- 20001544 TI - The University of British Columbia model of interprofessional education. AB - The College of Health Disciplines, at the University of British Columbia (UBC) has a long history of developing interprofessional learning opportunities for students and practitioners. Historically, many of the courses and programmes were developed because they intuitively made sense or because certain streams of funding were available at particular times. While each of them fit generally within our understanding of interprofessional education in the health and human service education programs, they were not systematically developed within an educational or theoretical framework. This paper discusses the model we have subsequently developed at the College for conceptualizing the various types of interprofessional experiences offered at UBC. It has been developed so that we can offer the broadest range of courses and most effective learning experiences for our students. Our model is based on the premise that there are optimal learning times for health and human services students (and practitioners) depending upon their stage of development as professionals in their respective disciplines and their readiness to learn and develop new perspectives on professional interaction. PMID- 20001545 TI - Multidisciplinary collaboration as a loosely coupled system: integrating and blocking professional boundaries with objects. AB - In this paper, the concept of loosely coupled systems is applied to describe multidisciplinary collaboration in healthcare. It is further argued that tools employed in collaborative activities may be regarded as object components of such a system. Drawing on observational studies and interviews of a group of health professionals from different disciplines collaborating on breast cancer, it is argued that differences in use of such objects may either inhibit or encourage cross-boundary collaboration. This effect is influenced by how the meanings of these objects vary between participants in the collaborative interaction. Meaning variation allows for more integration across boundaries, while meaning immutability may block the same boundaries. This finding is important for multidisciplinary contexts, adding new knowledge to the important quest for integrating relationships across professional boundaries. PMID- 20001546 TI - Stereotyping at the undergraduate level revealed during interprofessional learning between future doctors and biomedical scientists. AB - Interprofessional education (IPE) involving undergraduate health professionals is expected to promote collaboration in their later careers. The role of IPE between doctors and biomedical scientists has not been explored at the undergraduate level. Our aim was to introduce IPE sessions for medical and biomedical students in order to identify the benefits and barriers to these groups learning together. Medical and biomedical students together discussed laboratory results, relevant literature, and ideas for developing new diagnostic tools. The programme was evaluated with questionnaires and interviews. While there was general support for the idea of IPE, medical and biomedical students responded differently. Biomedical students were more critical, wanted more explicit learning objectives and felt that their professional role was often misunderstood. The medical students were more enthusiastic but regarded the way the biomedical students communicated concerns about their perceived role as a barrier to effective interprofessional learning. We conclude that stereotyping, which can impede effective collaborations between doctors and biomedical scientists, is already present at the undergraduate level and may be a barrier to IPE. Effective learning opportunities should be supported at the curriculum level and be designed to specifically enable a broad appreciation of each other's future professional roles. PMID- 20001547 TI - Wound care in primary health care: district nurses' needs for co-operation and well-functioning organization. AB - Most patients with leg- and foot ulcers are managed within non-institutional care. The aim of this study was to investigate the district nurses' wound management, including wound appearance, assignment of responsibility, guidelines for wound treatment and co-operation with other professional groups. The study has a descriptive quantitative approach. Data was collected using a wound registration form and a questionnaire. The selection of participants was made by random sampling. District nurses (n = 26) in five health-care centers situated in central Stockholm and two of its suburbs, participated in the study. The results show that the wound appearance is dominated by traumatic wounds. Approximately 40% of the wounds were not medically diagnosed. The area of responsibility of different professional groups was not defined and guidelines for wound treatment were mostly lacking. The decision about wound management was generally made by the district nurse. Co-operation with the general practitioner was lacking and when a consultation with dermatologist was required, the routines concerning referral were undefined. Co-operation with the assistant nurses consisted of redressing the wounds in home care. Interprofessional co-operation was regarded as important for wound healing. The paper provides insights into the district nurses' wound management and co-operation in wound care. PMID- 20001552 TI - Discontinued drugs in 2008: anti-infectives. AB - Of the many drugs dropped from development in 2008, 27 were under development for therapy or prophylaxis of infectious diseases. The majority of these were for diseases of viral etiology (15 agents), while nine were antibacterials and the remaining three were directed against fungal and protozoan pathogens. The antiviral agents were primarily against HIV and hepatitis C virus, together with agents against hepatitis B virus and West Nile virus. The antibacterial agents were mostly from known classes (carbapenems, fluoroquinolones, oxazolidinones), but also included several novel compounds and approaches. With regard to drug type, 19 were small-molecule therapeutics, seven were biologics comprising five vaccines and two antibodies, and one was a high-molecular-weight polymer. Each of these agents is briefly reviewed and the reasons for failure are discussed. PMID- 20001553 TI - Polo-like kinase inhibitors: an emerging opportunity for cancer therapeutics. AB - IMPORTANCE OF THE FIELD: The Polo-like kinase (Plk) family has emerged as an important regulator in cell cycle progression. Plks belong to a family of serine/threonine kinases and exist in four isoforms Plk1- 4. However, only one of these isoforms, Plk1, is shown to be involved in the activation of Cdc2, chromosome segregation, centrosome maturation, bipolar spindle formation and execution of cytokinesis. The activity of Plk1 is elevated in tissues and cells with a high mitotic index. In patients, Plk1 is overexpressed in tumors including those derived from lung, breast, colon, pancreas, prostate and ovary. Plk1 depletion is associated with the decrease in cell viability and induction of apoptosis in various cancerous cells. Several Plk1 inhibitors are in different phases of clinical development for anticancer therapy. AREAS COVERED IN THIS REVIEW: The focus of present review is to highlight Plk1 as a promising therapeutic approach for the treatment of cancer. The review discusses the role of Plk1 in cancer and the current status of Plk1 inhibitors, as well as highlighting the possible beneficial effect of inhibition of Plk1 as compared to other mitotic targets. WHAT THE READER WILL GAIN: Readers will get a comprehensive overview of Plk1 as a novel anticancer drug target. This review will also update readers about the progress made in the field of Plk1 inhibitors. TAKE HOME MESSAGE: The current literature about Plk1 inhibitors and knockout studies favor Plk1 inhibition as a potential antitumor therapy. PMID- 20001554 TI - Targeting voltage-gated sodium channels for pain therapy. AB - Drugs inhibiting voltage-gated sodium channels have long been used as analgesics, beginning with the use of local anaesthetics for sensory blockade and then with the discovery that Nav-blocking anticonvulsants also have benefit for pain therapy. These drugs were discovered without knowledge of their molecular target, using traditional pharmacological methods, and their clinical utility is limited by relatively narrow therapeutic windows. Until recently, attempts to develop improved inhibitors using modern molecular-targeted screening approaches have met with limited success. However, in the last few years there has been renewed activity following the discovery of human Nav1.7 mutations that cause striking insensitivity to pain. Together with recent advances in the technologies required to prosecute ion channels as drug targets, this has led to significant progress being made. This article reviews these developments and summarises current findings with these emerging new Nav inhibitors, highlighting some of the unanswered questions and the challenges that remain before they can be developed for clinical use. PMID- 20001555 TI - Investigational drugs for hepatitis C. AB - IMPORTANCE OF THE FIELD: Hepatitis C virus (HCV) infection affects 180 million people worldwide. Standard anti-HCV therapy combines effect of pegylated IFN alpha on immune response and antiviral activity of ribavirin. Sustained virological response rate achieved with this standard of care medication is around 50% for HCV genotype 1, the most prevalent worldwide genotype. So there is an obvious need to enhance efficacy of treatment. AREAS COVERED IN THIS REVIEW: We describe novel treatment options studied within the recent few years, which are focused on inhibitors of HCV-specific enzymes such as NS3/4 protease and NS5B polymerase or influence host-virus interactions. WHAT THE READER WILL GAIN: According to the most recent data, triple therapies consisted of pegylated interferons, ribavirin and protease inhibitors demonstrate significant improvement of treatment efficacy in comparison to current standard of care medication. Moreover, new forms of interferons and ribavirin, more effective, less toxic, and more convenient, become probably a fundamental component of anti HCV therapy in the near future. TAKE HOME MESSAGE: In coming years, we can expect that triple therapy becomes a standard medication, and treatment without interferon and/or ribavirin becomes a new studied therapeutic scenario. PMID- 20001557 TI - Investigational drugs for allergic rhinitis. AB - Allergic rhinitis (AR) is a high-prevalence disease, triggered by an IgE-mediated reaction, and sustained by a complex inflammatory network of cells, mediators, and cytokines. When the exposure to allergens persists, the inflammatory process becomes chronic. The current therapeutic strategy is based on allergen avoidance when possible, drugs and allergen immunotherapy. The main drugs are oral and topical antihistamines and nasal steroids. They are overall effective in controlling symptoms, but do not modify the immune background that leads to allergic inflammation. In addition, safety concerns may be present, especially for prolonged treatments. Thus, efforts are currently made to improve the existing molecules and to develop new drugs, in order to achieve greater clinical efficacy with a better tolerability. Also, attempts are made to selectively block relevant signal pathways of the allergic reaction by means of specific anti mediators. Specific immunotherapy, in addition to the clinical effect, is capable of modifying the Th2-biased immune response. Thus, an intense research activity is presently ongoing with the aim of improving the characteristics and modes of action of this treatment. PMID- 20001556 TI - Targeting the transforming growth factor-beta signaling pathway in human cancer. AB - The transforming growth factor-ss (TGF-beta) signaling pathway plays a pivotal role in diverse cellular processes. TGF-beta switches its role from a tumor suppressor in normal or dysplastic cells to a tumor promoter in advanced cancers. It is widely believed that the Smad-dependent pathway is involved in TGF-beta tumor-suppressive functions, whereas activation of Smad-independent pathways, coupled with the loss of tumor-suppressor functions of TGF-beta, is important for its pro-oncogenic functions. TGF-beta signaling has been considered a useful therapeutic target. The discovery of oncogenic actions of TGF-beta has generated a great deal of enthusiasm for developing TGF-beta signaling inhibitors for the treatment of cancer. The challenge is to identify the group of patients where targeted tumors are not only refractory to TGF-beta-induced tumor suppressor functions but also responsive to the tumor-promoting effects of TGF-beta. TGF beta pathway inhibitors, including small and large molecules, have now entered clinical trials. Preclinical studies with these inhibitors have shown promise in a variety of different tumor models. Here, we focus on the mechanisms of signaling and specific targets of the TGF-beta pathway that are critical effectors of tumor progression and invasion. This report also examines the therapeutic intervention of TGF-ss signaling in human cancers. PMID- 20001558 TI - Novel TNF antagonists for the treatment of rheumatoid arthritis. AB - IMPORTANCE OF THE FIELD: TNF antagonists have become an integral part of the standard of care for patients with rheumatoid arthritis (RA). Two additional TNF antagonists have recently been approved in the US for this indication, and clinicians will need to understand the data on the efficacy and safety of these agents in order to properly place them into the clinical treatment algorithm. AREAS COVERED IN THIS REVIEW: This review covers the published clinical trial data on the use of certolizumab and golimumab for RA. The literature search included manuscripts published through September 2009 and abstracts from major meetings (ACR and EULAR) in 2007 - 2009. WHAT THE READER WILL GAIN: This paper will review the efficacy and safety of golimumab and certolizumab in the treatment of RA at various stages of disease. Use of these agents will be described in the context of other available alternatives, including other TNF antagonists and other biologic therapies. TAKE HOME MESSAGE: Certolizumab and golimumab have comparable efficacy and safety profiles compared with previously approved TNF antagonists. These two agents have several unique theoretical benefits when compared to existing agents, but the available published data do not suggest a clear clinical advantage at this time. PMID- 20001559 TI - Targeting cancer with phosphodiesterase inhibitors. AB - IMPORTANCE OF THE FIELD: For many cancers, there has been a shift from management with traditional, nonspecific cytotoxic chemotherapies to treatment with molecule specific targeted therapies that are used either alone or in combination with traditional chemotherapy and radiation therapy. Accumulating data suggest that multi-targeted agents may produce greater benefits than those observed with single-targeted therapies, may have acceptable tolerability profiles, and may be active against a broader range of tumour types. Thus, regulation of cyclic nucleotide signalling is properly regarded as a composite of multiple component pathways involved in diverse aspects of tumour cell function. The impairment of cAMP and/or cGMP generation by overexpression of PDE isoforms that has been described in various cancer pathologies, and the effects of PDE inhibitors in tumour models in vitro and in vivo, may offer promising insight into future cancer treatments because of the numerous advantages of PDE inhibitors. AREAS COVERED IN THIS REVIEW: In this review, we focus on the expression and regulation of cyclic nucleotide phosphodiesterases (PDEs) in tumour progression and provide evidence that PDE inhibitors may be effective agents for treating cancer; the review covers literature from the past several years. WHAT THE READER WILL GAIN: PDEs have been studied in a variety of tumours; data have suggested that the levels of PDE activity are elevated and, therefore, the ratio of cGMP to cAMP is affected. In addition, PDE inhibitors may be potential targets for tumour cell growth inhibition and induction of apoptosis. This review explores the prospects of targeting PDEs with therapeutic agents for cancer, as well as the shortcomings of this approach such as dose-limiting side effects, toxicity/efficacy ratio and selectivity towards tumour tissue. In addition, it includes opinions and suggestion for developing PDE inhibition for cancer treatment from initial concept to potential therapeutic application and final relevance in clinical use. TAKE HOME MESSAGE: Impaired cAMP and/or cGMP generation upon overexpression of PDE isoforms has been described in various cancer pathologies. Inhibition of selective PDE isoforms, which raises the levels of intracellular cAMP and/or cGMP, induces apoptosis and cell cycle arrest in a broad spectrum of tumour cells and regulates the tumour microenvironment. Therefore, the development and clinical application of inhibitors specific for individual PDE isoenzymes may selectively restore normal intracellular signalling, providing antitumour therapy with reduced adverse effects. PMID- 20001560 TI - The efficacy and safety of telaprevir - a new protease inhibitor against hepatitis C virus. AB - IMPORTANCE OF THE FIELD: Hepatitis C virus (HCV) is the main agent of chronic hepatitis, liver cirrhosis and hepatocellular carcinoma in the western world. It affects more than 170 million people worldwide. HCV treatment, based on the combination of Peg-interferon and ribavirin, is effective in about 50% of treated patients. Therefore, there is a need to develop new drugs active against HCV. AREAS COVERED IN THIS REVIEW: Data were obtained by searching for all full articles on Medline and abstracts presented at major international congresses on viral hepatitis. WHAT THE READER WILL GAIN: A review of clinical data about the efficacy and safety of telaprevir (VX-950), the HCV protease inhibitor that is in the most advanced phase of clinical development. TAKE HOME MESSAGE: Telaprevir has an acceptable pharmacokinetic profile and seems to be a potent antiviral drug against HCV, although, owing to a low genetic barrier, resistant variants emerge within a few days when used in monotherapy, thereby decreasing its efficacy. Consequently, telaprevir has been combined with pegylated-interferon and ribavirin in clinical trials. This triple combination is more effective but has a higher rate of adverse events (notably rash) than the standard of care, despite the shorter duration of therapy. PMID- 20001561 TI - Darapladib. AB - IMPORTANCE OF THE FIELD: Atherosclerosis is an inflammatory-immune mediated disease process. Plaque rupture is responsible for the clinical events of ischemic death, myocardial infarction, acute coronary syndromes and ischemic strokes. Lipoprotein-associated phospholipase A(2) (Lp-PLA(2)) seems to play a major role in the development of such high-risk lesions, in both the coronary and carotid arteries. Darapladib is a selective inhibitor of Lp-PLA(2). AREAS COVERED IN THIS REVIEW: An overview of darapladib by reviewing the studies (1990 - 2009) that have provided the rationale for the development of darapladib; and a discussion of its potential merit as a new therapeutic drug to target high-risk atherosclerosis. WHAT THE READER WILL GAIN: The reader should gain an understanding of the importance of inflammation during atherogenesis as well as of the biology of Lp-PLA(2) and its proatherogenic role. Additional insights will be gained into the role of selective inhibitors of Lp-PLA(2) as new therapeutic agents. TAKE HOME MESSAGE: Darapladib is a selective inhibitor of Lp-PLA(2) and represents a new class of therapeutic agents that target inflammation to treat high-risk atherosclerosis. PMID- 20001562 TI - HER-dimerization inhibitors: evaluating pertuzumab in women's cancers. AB - IMPORTANCE OF THE FIELD: Pertuzumab is a human EGF receptor (HER)-dimerization inhibitor that represent a novel class of agents aimed at blocking HER2 from pairing with other receptors of the HER family. In this review, we discuss the background and scientific rationale, related to pertuzumab as it has undergone development for women's cancers. AREAS COVERED IN THIS REVIEW: Pre-clinical and clinical trials, published or presented at national meetings from 1995 to the present, are included in this review. WHAT THE READER WILL GAIN: A Phase II trial in HER2-positive metastatic breast cancer showed promising activity of pertuzumab when it was combined with trastuzumab and a randomized Phase III trial is now underway. Pertuzumab, when evaluated in recurrent ovarian cancer showed limited activity when combined with chemotherapy in platinum-sensitive and platinum resistant disease. However, a recent subset analysis suggests that HER3 mRNA levels may predict response to pertuzumab in ovarian cancer. TAKE HOME MESSAGE: HER-dimerization inhibitors represent a novel mechanism of inhibition of HER pathways. Pertuzumab may play a role in the management of HER2-positive breast cancer. The potential benefit of pertuzumab in ovarian cancer is unclear, but low HER3 levels may suggest a sub-group of patients that can benefit from pertuzumab. PMID- 20001564 TI - Does the reproductive cycle influence sleep patterns in women with sleep complaints? AB - OBJECTIVE: The aim of the present study was to investigate how prominent gynecological factors (regular/irregular menstrual cycle, premenstrual complaints, or menopause) can influence both subjective and objective sleep data. METHODS: A total of 931 women who sought clinical assistance because of a sleep complaint were included in the investigation. All subjects filled out a sleep and gynecological questionnaire prior to undergoing a polysomnography (PSG) recording. RESULTS: Premenopausal women with an irregular menstrual cycle were more likely to report sleep difficulties (related to falling sleep and insomnia symptoms) than those with a regular cycle (odds ratio 2.1; 95% confidence interval 1.2-3.5). These women showed increased light sleep stages and awakenings. Premenstrual complaints were reported by a higher percentage of women with periodic leg movement, lower time in saturation under 90%, and higher sleep efficiency. A lower percentage of women who took hormonal contraceptives reported snoring, had fewer arousals and longer REM latency compared to those not taking hormonal contraceptives. Menopausal women with hot flushes had more restless leg complaints. Awake time was shorter in hormone therapy users compared with non users. Women with menopause are more likely to have an apnea-hypoapnea index greater than 5/h. CONCLUSION: The present findings suggest that gynecological status is associated with subjective sleep quality and objective sleep parameters in women with sleep complaints. PMID- 20001565 TI - Menopause management: a cardiovascular risk-based approach. AB - Hormone replacement therapy (HRT) remains the gold standard for the management of menopausal symptoms; however, HRT use has declined due to concerns over possible adverse side-effects. Approaches to menopause management are continually being revised and these extend beyond the control of recognized menopausal symptoms to encompass wider aspects of menopausal women's health. Hypertension and associated cardiovascular risk are particularly important unmet needs in postmenopausal women, especially in the Asia-Pacific region which has a rapidly aging population and bears around half of the global burden of cardiovascular disease, two-thirds of which has been attributed to elevated blood pressure. As first point of contact for women with menopausal symptoms, gynecologists play a gatekeeper role in assessing women's health, providing appropriate lifestyle counseling, and, where appropriate, implementing treatment or referral to relevant specialists. This paper, with contributions by gynecologists and cardiologists from Asia Pacific and beyond, summarizes available evidence and provides a treatment algorithm that employs a flexible blood pressure classification strategy to assist physicians in their decision-making for the individualized management of menopausal symptoms in women with low, moderate and high cardiovascular risk, and also for women with diabetes. Individualized HRT according to cardiovascular risk may yield improvements in cardiovascular health, as well as managing menopausal symptoms. PMID- 20001563 TI - Optimal tolerability of ultra-low-dose continuous combined 17beta-estradiol and norethisterone acetate: laboratory and safety results. AB - OBJECTIVE: To evaluate the influence of two ultra-low doses of oral continuous combined hormone therapy and placebo on metabolic parameters, and to assess safety endpoints and overall tolerability in healthy postmenopausal women. DESIGN: In a subpopulation of the Clinical study on Hormone dose Optimisation In Climacteric symptoms Evaluation (CHOICE) trial, lipids and parameters of glucose metabolism and hemostasis were analyzed in Nordic women (n = 158) at baseline and after 12 and 24 weeks of treatment with 0.5 mg 17beta-estradiol (E2) + 0.25 mg norethisterone acetate (NETA), 0.5 mg E2 + 0.1 mg NETA or placebo. Adverse events occurring from the first trial-related activity, whether related or not related to the study medication, were recorded for the entire population (n = 575) of the trial. The seriousness, relationship to treatment and the reason for withdrawal were reported. RESULTS: Both ultra-low-dose combinations were neutral to changes in lipid and lipoprotein, hemostasis parameters and carbohydrate metabolism during the trial. The incidence of serious adverse events was only 1% in respective treatment groups. Adverse events were the reason for withdrawal in only 2% and 6% of women in the 0.5 mg E2 + 0.25 mg and 0.1 mg NETA groups, and in 8% in the placebo group. No weight gain or change in blood pressure was reported during the trial in any of the study groups. CONCLUSION: The treatments had neutral effects on metabolic parameters in the study population. Excellent tolerability of both ultra-low doses resulted in high completion rates. PMID- 20001566 TI - Ankle stiffness in postmenopausal women: influence of hormone therapy and menopause nature. AB - BACKGROUND: Literature suggests that muscle-tendon unit (MTU) stiffness is implicated in human motion performance and injuries. In addition, muscle and joint pains comprise a set of very common climacteric symptoms. Concomitant with such symptoms, physical fragility and mobility limitations are expected, which further affect the functional physical fitness of postmenopausal women. To the authors' knowledge, this is the first study relating MTU stiffness with hormone therapy (HT) and the nature of menopause. The purpose of this study was to investigate the influence of HT and the nature of menopause on triceps-surae MTU stiffness in postmenopausal women. METHODS: Ninety-three women participated in this study. The data concerning menopause were obtained through medical consultation. MTU stiffness was assessed in vivo using a damped oscillation technique and a load equivalent to 30% of maximal voluntary isometric contraction. RESULTS: None of the following pair groups showed statistical differences for MTU stiffness: the HT group (19,426 +/- 4148 N/m) and the without HT group (20,056 +/- 3579 N/m); the natural menopause group (19,525 +/- 3718 N/m) and the induced menopause group (20,469 +/- 4705 N/m). No significant differences were also found between the following pair groups: the natural menopause with-HT group (19,078 +/- 3910 N/m) and the without-HT group (20,076 +/- 3442 N/m); the induced menopause with-HT group (20,756 +/- 4932 N/m) and the without-HT group (19,942 +/- 4656 N/m). CONCLUSION: The data suggest that MTU stiffness is not related to either the administration of HT in postmenopausal women or to the nature of menopause. PMID- 20001567 TI - Changing the dose metric for inhalation toxicity studies: short-term study in rats with engineered aerosolized amorphous silica nanoparticles. AB - Inhalation toxicity and exposure assessment studies for nonfibrous particulates have traditionally been conducted using particle mass measurements as the preferred dose metric (i.e., mg or microg/m(3)). However, currently there is a debate regarding the appropriate dose metric for nanoparticle exposure assessment studies in the workplace. The objectives of this study were to characterize aerosol exposures and toxicity in rats of freshly generated amorphous silica (AS) nanoparticles using particle number dose metrics (3.7 x 10(7) or 1.8 x 10(8) particles/cm(3)) for 1- or 3-day exposures. In addition, the role of particle size (d(50) = 37 or 83 nm) on pulmonary toxicity and genotoxicity endpoints was assessed at several postexposure time points. A nanoparticle reactor capable of producing, de novo synthesized, aerosolized amorphous silica nanoparticles for inhalation toxicity studies was developed for this study. SiO(2) aerosol nanoparticle synthesis occurred via thermal decomposition of tetraethylorthosilicate (TEOS). The reactor was designed to produce aerosolized nanoparticles at two different particle size ranges, namely d(50) = approximately 30 nm and d(50) = approximately 80 nm; at particle concentrations ranging from 10(7) to 10(8) particles/cm(3). AS particle aerosol concentrations were consistently generated by the reactor. One- or 3-day aerosol exposures produced no significant pulmonary inflammatory, genotoxic, or adverse lung histopathological effects in rats exposed to very high particle numbers corresponding to a range of mass concentrations (1.8 or 86 mg/m(3)). Although the present study was a short-term effort, the methodology described herein can be utilized for longer-term inhalation toxicity studies in rats such as 28-day or 90 day studies. The expansion of the concept to subchronic studies is practical, due, in part, to the consistency of the nanoparticle generation method. PMID- 20001568 TI - Bone resorption acceleration and calcium reabsorption impairment in a Thai population with high cadmium exposure. AB - Some residents of the Mae Sot district in Thailand have suffered long-term exposure to elevated dietary levels of cadmium. To test the hypothesis that chronic dietary cadmium exposure can cause imbalance in calcium dynamics and accelerate bone resorption, a group of these residents (156 men and 256 women aged >/= 50) were selected on the basis of previous records of elevated urinary cadmium and tested for urinary and blood cadmium, bone formation and resorption markers, and the renal tubular dysfunction markers. Both genders had high levels of blood and urinary cadmium and high urinary levels of the markers for renal dysfunction and bone resorption in a dose-response relationship to urinary cadmium. The excretion of bone resorption markers was positively correlated to the ratio of excreted calcium and urinary cadmium. The results of a multivariate regression analysis indicated that bone resorption was accelerated by impaired calcium reabsorption in renal tubules. PMID- 20001569 TI - Management of severe hypoglycaemia: cultural similarities, differences and resource consumption in three European countries. AB - BACKGROUND: To investigate the characteristics of people with insulin-treated diabetes, who have experienced severe hypoglycaemic events (SHEs), in Germany, Spain or UK. METHODS: Patients with type 1 (n=319) or insulin-treated type 2 diabetes (n=320) who had experienced >= 1 SHE in the preceding year were enrolled. Their median age was 53 years (range, 16-94 years). Data were collected using a questionnaire administered by an experienced interviewer. RESULTS: The median number of reported SHEs was 2-3 in 12 months. Most events (69%) occurred at home, usually during the day or evening (74%) and most commonly due to insufficient food consumption (45%). In patients whose hypoglycaemia awareness was tested, 68% had normal awareness. Patients requiring emergency healthcare treatment frequently had impaired hypoglycaemia awareness, and developed hypoglycaemic coma more often. Hospital treatment was usually provided in an emergency department (72-94%). The duration of stay was longest in Germany. Following a SHE, patients receiving professional treatment were more likely to: consult their physician, test their blood glucose more often, adjust insulin dose and receive self-management training. CONCLUSIONS: This survey of diabetes patients aged 16-94 years showed that SHEs represent a substantial burden on national healthcare systems in Germany, UK and Spain. The pattern of occurrence and treatment was similar in all three countries, despite differences in cultures and healthcare systems. PMID- 20001570 TI - Costs of managing severe hypoglycaemia in three European countries. AB - OBJECTIVES: To assess the costs of severe hypoglycaemic events (SHEs) in diabetes patients in Germany, Spain and the UK. METHODS: Healthcare resource use was measured by surveying 639 patients aged >= 16 years, receiving insulin for type 1 (n=319) or type 2 diabetes (n=320), who experienced >= 1 SHE in the preceding year. Patients were grouped by location of SHE treatment: group 1, community (family/domestic); group 2, community (healthcare professional); group 3, hospital. Costs were calculated from published unit costs applied to estimated resource use. Costs per SHE were derived from patient numbers per subgroup. Weighted average costs were derived using a prevalence database. RESULTS: Hospital treatment was a major cost in all countries. In Germany and Spain, costs per SHE for type 1 patients differed from those for type 2 patients in each group. Average SHE treatment costs were higher for patients with type 2 diabetes (Germany, ?533; Spain, ?691; UK, ?537) than type 1 diabetes patients (?441, ?577 and ?236, respectively). Telephone calls, visits to doctors, blood glucose monitoring and patient education contributed substantially to costs for non hospitalised patients. CONCLUSIONS: Treatment of SHEs adds significantly to healthcare costs. Average costs were lower for type 1 than for insulin-treated type 2 diabetes, in all three countries. PMID- 20001571 TI - Endocrine factors associated with non-alcoholic fatty liver disease in women with polycystic ovary syndrome: do androgens play a role? AB - OBJECTIVE: To characterise the metabolic profile of women with polycystic ovary syndrome (PCOS) and non-alcoholic fatty liver disease (NAFLD) and to determine whether circulating androgens differ in PCOS women with NAFLD compared to PCOS subjects without NAFLD. METHODS: Retrospective study of 21 women with PCOS, elevated liver enzymes and ultrasound evidence of hepatic steatosis matched with 32 PCOS women with normal liver enzymes. Extensive demographic, endocrine and metabolic data were compared. Pearson's correlation coefficients were calculated to assess for potential relationships between the free androgen index (FAI) and other dependent variables. RESULTS: PCOS subjects with NAFLD demonstrate greater insulin resistance but have similar circulating androgen levels. CONCLUSION: In this pilot study, insulin resistance was the most prominent feature characterising NAFLD complicating PCOS. Total testosterone, FAI, DHEAS and 17 hydroxyprogesterone levels were similar between patients with PCOS and without NAFLD. PMID- 20001572 TI - Spontaneous subcapsular hematoma of liver in pregnancy of unknown etiology- conservative management: a case report. AB - Mrs. AB, a 40-year-old woman, in her second pregnancy had a spontaneous hematoma of liver of unknown etiology that was managed successfully conservatively under the umbrella of the multidisciplinary care. The subcapsular hematoma was diagnosed at 31 weeks gestational age while she was being investigated because of sudden and gross drop of hemoglobin from 12.8 to 8 g/dl in 2 weeks duration. The dilemma remains as how to manage her future pregnancies and what are the risks of recurrence. PMID- 20001573 TI - Withdrawn: Enhancing efficacy and mucosa-tropic distribution of an oral HIV-PsV DNA vaccine in animal models. PMID- 20001574 TI - Efficacy and safety of linezolid in methicillin-resistant Staphylococcus aureus (MRSA) complicated skin and soft tissue infection (cSSTI): a meta-analysis. AB - OBJECTIVE: To evaluate the clinical and microbiological outcomes of linezolid versus vancomycin in methicillin-resistant Staphylococcus aureus (MRSA) complicated skin and soft-tissue infection (cSSTI) using a meta-analysis. RESEARCH DESIGN AND METHODS: Clinical trials were identified using PubMed, the Cochrane Central Register of Controlled Trials, and the International Pharmaceutical Abstracts from inception to March 2009. Primary outcomes evaluated resolution of signs and symptoms of infection in clinically evaluable (CE) patients, and microbiological eradication in both the modified intent-to-treat (MITT) and MRSA evaluable (MRSA ME) patients. MITT patients had a culture confirmed Gram-positive pathogen (S. aureus) at baseline. Secondary outcomes included mortality, adverse drug reactions (ADR), and discontinuation due to ADRs. The Mantel-Haenszel odds ratios (OR) with 95% confidence intervals (CI) were calculated using the fixed effects model based on the assumption that there was little to no heterogeneity between the studies in the primary analysis. Sensitivity analyses were performed for each outcome by removing the most weighted study. RESULTS: Five studies with a total of 2652 patients (1361/linezolid; 1291/vancomycin) were identified. Resolution of infection in CE patients (OR = 1.41; 95% CI: 1.03, 1.95) and MITT patients (OR = 1.91; 95% CI: 1.33, 2.76) favored the use of linezolid over vancomycin, but did not remain significant after sensitivity analyses (CE: OR = 1.29; 95% CI: 0.81, 2.05; MITT: OR = 1.73; 95% CI: 0.87, 3.41). Microbiological eradication in MRSA ME patients consistently favored the use of linezolid over vancomycin (OR = 2.90; 95% CI: 1.90, 4.41). No difference in mortality was observed between the two groups (OR = 1.17; 95% CI: 0.85, 1.62). Higher proportions of linezolid patients were found to have diarrhea (119/1361 vs. 52/1291), nausea (102/1361 vs. 46/1291) and thrombocytopenia (54/1121 vs. 8/1071), whereas a higher proportion of vancomycin patients were found to have renal insufficiency compared to linezolid (16/634 vs. 4/703). Inconsistent results were seen with the incidence of anemia and rash between the base-case (anemia: OR = 1.36; 95% CI: 0.90, 2.08; rash: OR = 0.26; 95% CI: 0.10, 0.68) and sensitivity analyses (anemia: OR = 2.33; 95% CI: 1.24, 4.37; rash: OR = 0.57; 95% CI: 0.12, 2.71). Discontinuation due to ADRs was not statistically different between linezolid and vancomycin (OR = 1.06; 95% CI: 0.75, 1.51). CONCLUSION: Resolution of infection in CE and MITT patients were inconsistent; however, a sub-analysis revealed that linezolid was more likely to consistently achieve microbiologic eradication in MRSA ME patients. Apparent risks of thrombocytopenia, nausea, diarrhea, and possibly anemia may limit linezolid use in treating MRSA cSSTI. This study was limited due to an inability to assess for the effects of hetero-resistance and appropriate vancomycin dosing on outcomes. Moreover, the small number of studies made controlling for heterogeneity challenging. PMID- 20001575 TI - Evaluating patient satisfaction with specific migraine therapy based on initial treatment expectations: the PAX study. AB - AIMS AND OBJECTIVES: The PAX study was a naturalistic, prospective survey evaluating treatment expectations and satisfaction in a typical French migraine population. METHODS: A total of 1710 patients were recruited by 489 general practitioners across France. Despite a high drop-out rate (due to one or more deviations from protocol and/or missing data), the analysable per-protocol population (PPP) patients (n = 615) remained representative with regard to the overall migraine population. Patients ranked expectations according to four criteria at baseline, and were asked to report satisfaction for each criterion at baseline and for six consecutive attacks. The highest-ranked treatment expectation was rapid relief of headache, followed by pain-free response, relief of associated symptoms, and ability to carry on present activity, respectively. RESULTS: At inclusion, >50% of patients were using non-specific medications (analgesics, NSAIDs), and 30% reported global satisfaction with treatment. At the end of the survey, >75% of patients were using specific medications (predominantly triptans), and global treatment satisfaction increased to 83%, independent of treatment expectation rankings at baseline. CONCLUSIONS: These survey results suggest that satisfaction with acute migraine treatment increases when specific medications are prescribed, irrespective of which treatment expectation is considered most important at baseline. This emphasizes the need for improvements in the management of migraine, to ensure that optimal treatment is being provided with regard to pharmacological intervention. PMID- 20001576 TI - Meta-analysis of the evidence for a partially hydrolyzed 100% whey formula for the prevention of allergic diseases. AB - OBJECTIVE: Infants with a documented hereditary risk of atopy (i.e., an affected parent and/or sibling) who cannot be breastfed exclusively are recommended to receive a formula with confirmed reduced allergenicity, i.e., a partially or extensively hydrolyzed formula (pHF and eHF, respectively), as a means of preventing allergic reactions. The efficacy of each hydrolyzed formula for the prevention of allergic diseases should be established separately, as factors such as the protein source, hydrolysis method and degree of hydrolysis that often depend on the manufacturer contribute to differences among hydrolysates. The aim was to systematically review data on the efficacy of a partially hydrolyzed 100% whey formula (pHF) in reducing the risk of allergy in healthy infants at high risk for allergy. METHODS: The Cochrane Library, MEDLINE, EMBASE, and CINAHL databases were searched in September 2009 (from inception to September 2009) for randomized and quasi-randomized controlled trials (RCTs); additional references were obtained from reviewed articles. The company that manufactures the pHF used was contacted for unpublished data. RESULTS: The search yielded 84 citations. Fifteen RCTs were included, some of which had potential methodological limitations such as unclear or inadequate allocation concealment, no intention-to treat analysis, and no true blinding. For primary outcomes, i.e., all allergic diseases and atopic eczema/atopic dermatitis, use of the pHF compared with standard formula (SF) was associated with reduced risks (incidence, cumulative incidence, period prevalence) that were statistically significant for most, albeit not all, time points. Comparison of groups who received the pHF versus extensively hydrolyzed (eH) whey formula revealed no significant differences in outcomes except for reductions in the cumulative incidences of all allergic diseases at 0 to 36 months of age. Comparison of groups who received the pHF versus eH casein formula revealed no significant difference in outcomes between groups. CONCLUSIONS: The use of the pHF compared to SF is effective in allergy prevention in children at high risk for allergy at most time points. These results should be interpreted with caution due to a lack of methodological rigor in many trials. Reassuringly, the strongest evidence comes from a well-designed and conducted, independently funded RCT. PMID- 20001577 TI - Comment on 'Huntington's disease presenting as ALS'. PMID- 20001578 TI - The importance of flower cells for the early diagnosis of acute adult T-cell leukemia/lymphoma with skin involvement. PMID- 20001579 TI - Genetic information nondiscrimination act insurance protections issued. PMID- 20001580 TI - Cancer genetic risk assessment and referral patterns in primary care. AB - PURPOSE: This study was undertaken to describe cancer risk assessment practices among primary care providers (PCPs). METHODS: An electronic survey was sent to PCPs affiliated with a single insurance carrier. Demographic and practice characteristics associated with cancer genetic risk assessment and testing activities were described. Latent class analysis supported by likelihood ratio tests was used to define PCP profiles with respect to the level of engagement in genetic risk assessment and referral activity based on demographic and practice characteristics. RESULTS: 860 physicians responded to the survey (39% family practice, 29% internal medicine, 22% obstetrics/gynecology (OB/GYN), 10% other). Most respondents (83%) reported that they routinely assess hereditary cancer risk; however, only 33% reported that they take a full, three-generation pedigree for risk assessment. OB/GYN specialty, female gender, and physician access to a genetic counselor were independent predictors of referral to cancer genetics specialists. Three profiles of PCPs, based upon referral practice and extent of involvement in genetics evaluation, were defined. CONCLUSION: Profiles of physician characteristics associated with varying levels of engagement with cancer genetic risk assessment and testing can be identified. These profiles may ultimately be useful in targeting decision support tools and services. PMID- 20001581 TI - Quantitative real-time polymerase chain reaction for the verification of genomic imbalances detected by microarray-based comparative genomic hybridization. AB - The American College of Medical Genetics guidelines for microarray analysis for constitutional cytogenetic abnormalities require abnormal or ambiguous results from microarray-based comparative genomic hybridization (aCGH) analysis be confirmed by an alternative method. We employed quantitative real-time polymerase chain reaction (qPCR) technology using SYBR Green I reagents for confirmation of 93 abnormal aCGH results (50 deletions and 43 duplications) and 54 parental samples. A novel qPCR protocol using DNA sequences coding for X-linked lethal diseases in males for designing reference primers was established. Of the 81 sets of test primers used for confirmation of 93 abnormal copy number variants (CNVs) in 80 patients, 71 sets worked after the initial primer design (88%), 9 sets were redesigned once, and 1 set twice because of poor amplification. Fifty-four parental samples were tested using 33 sets of test primers to follow up 34 CNVs in 30 patients. Nineteen CNVs were confirmed as inherited, 13 were negative in both parents, and 2 were inconclusive due to a negative result in a single parent. The qPCR assessment clarified aCGH results in two cases and corrected a fluorescence in situ hybridization result in one case. Our data illustrate that qPCR methodology using SYBR Green I reagents is accurate, highly sensitive, specific, rapid, and cost-effective for verification of chromosomal imbalances detected by aCGH in the clinical setting. PMID- 20001582 TI - Comparative study of three PCR-based copy number variant approaches, CFMSA, M PCR, and MLPA, in 22q11.2 deletion syndrome. AB - Small submicroscopic DNA copy number variants represent an important source of variation in the human genome, human phenotypic diversity, and disease susceptibility. Consequently, there is a pressing need for the development of methods allowing the efficient, accurate, and cheap measurement of genomic copy number polymorphisms in clinical cohorts. The PCR-based strategies, being cost effective and sensitive, are considered important in the development of screening techniques. PCR-based techniques such as multiplex PCR; multiplex ligation dependent probe amplification; and a new single-tube assay technique, the competitive fluorescent multiplex STRP assay, have been applied to 22q11.2 detection, a typical example of deletion syndromes. In this study, we compared the reliability and application of these three techniques in a cohort of 17 patients affected with 22q11.2 deletion and 300 normal controls. All three techniques shared 100% sensitivity; however, the competitive fluorescent multiplex STRP assay had the lowest possibility of concurrent false-positive signals from two adjoining probes in a genomic region. Moreover, it is a relatively fast and low-cost procedure to detect the deletion of 22q11.2 in numerous patients with several minor symptoms of deletion syndromes. Multiplex PCR, a rapidly developing and cheap technique, allows detection of atypical deletions. PMID- 20001584 TI - The influence of alcohol on behavioral recovery after mTBI in mice. AB - In the United States 258,000 people were injured in 2004 in motor vehicle accidents that were caused by drivers under the influence of alcohol. The majority of these drivers were binge drinkers, most notably young people who tend to drink heavily during the weekends, but rarely drink alcohol during the week. Since a large proportion of the injuries involved head injuries, the present study aimed at investigating the influence of binge alcohol drinking on mild traumatic brain injury (mTBI) in an animal model. Mice had access to 0%, 7.5%, 15%, or 30% alcohol solutions for 48 consecutive hours once a week for 4 weeks as the sole source of fluids (the remaining time they drank water). Three experiments were done. For the first one (alcohol-mTBI-alcohol) the animals were subjected to a controlled mTBI injury by applying a closed-head weight drop, or a sham procedure. After the mTBI/sham-mTBI the animals got alcohol and /water for the same regimen for 4 additional weeks. In the second experiment (alcohol only) after the 4 weeks of drinking blood samples were collected, at the same time as the animals that underwent sham-mTBI or mTBI procedures. In the third experiment (mTBI-alcohol) the mice were subjected to mTBI/sham-mTBI without any treatment, and after mTBI they had alcohol for 4 weeks in the same regimen as in the previous experiments. At the end of the pharmacological treatment all animals were assessed using different behavioral tests. mTBI mice exhibited lower memory ability in the Y-maze, higher anxiety in the elevated plus maze, and lower retention in the passive avoidance test than sham-mTBI animals. Alcohol reversed these effects at all doses. The results suggest that alcohol drinking before trauma might have a protective effect on recovery from brain trauma, but not if consumed after the trauma. PMID- 20001585 TI - A case-comparison study of the subdural evacuating port system in treating chronic subdural hematomas. AB - OBJECT: The Subdural Evacuating Port System (SEPS) was recently introduced as a novel method of treating chronic subdural hematomas (SDHs). This system is a variation of the existing twist-drill craniostomy methods for treating chronic SDH. Compared with craniotomy or bur hole treatment of chronic SDH, this system offers the possibility of treatment at bedside without general anesthesia. In comparison with existing twist-drill methods, the system theoretically offers the advantage of a hermetically closed system that can evacuate a hematoma without an intracranial catheter. METHODS: The authors performed a case-control study of all chronic SDHs treated at a single institution over a 5-year period and compared the efficacy and safety of the SEPS to bur hole evacuation. Patients were matched for age, injury mechanism, medical comorbidities, use of anticoagulation, and radiographic appearance of the SDH. The primary outcome of interest was the recurrence rate in each group, which was evaluated by radiographic evidence as well as the number of patients requiring a second procedure. Secondary outcomes examined were mortality, infection, acute hematoma formation, seizure, length of hospital stay, length of intensive care unit stay, and discharge location. RESULTS: The authors found that there were no appreciable differences in symptoms on presentation, existing comorbidities, home medications, or laboratory values between the treatment groups. The average Hounsfield units of preoperative CT scanning was similar in both groups. Radiographic recurrence was statistically similar between the SEPS group (25.9%) and the bur hole group (18.5%; p = 0.37). Although there was a trend toward higher reoperation rates in the SEPS group, the need for a subsequent procedure was also statistically similar between the SEPS group (25.9%) and the bur hole group (14.8%; p = 0.25). The mortality rate was not significantly different between the SEPS group (9.5%) and the bur hole group (4.8%; p = 0.50). The SEPS procedure provided a mean reduction in SDH thickness of 27.3% compared with 37.9% with bur hole (p = 0.05) when comparing the preoperative CT scan with the first postoperative CT scan. The percentage of reduction in SDH thickness when comparing the preoperative CT scan with the most recent postoperative CT scan was 40.5% in the SEPS group and 45.4% in the bur hole group (p = 0.31). CONCLUSIONS: The SEPS offers an alternative type of twist drill craniostomy for the treatment of chronic SDH with a trend toward higher recurrence in our experience. The efficacy and safety of SEPS is similar to that of other twist-drill methods reported in the literature. In the authors' experience, the efficacy of this treatment as measured by radiographic worsening or the need for a subsequent procedure is statistically similar to that of bur hole treatment. There was no difference in mortality or other adverse outcomes associated with SEPS. PMID- 20001586 TI - Trans-sodium crocetinate enhancing survival and glioma response on magnetic resonance imaging to radiation and temozolomide. AB - OBJECT: Glioblastoma (GB) tumors typically exhibit regions of hypoxia. Hypoxic areas within the tumor can make tumor cells less sensitive to chemotherapy and radiation therapy. Trans-sodium crocetinate (TSC) has been shown to transiently increase oxygen to hypoxic brain tumors. The authors examined whether this improvement in intratumor oxygenation translates to a therapeutic advantage when delivering standard adjuvant treatment to GBs. METHODS: The authors used C6 glioma cells to create a hypoxic GB model. The C6 glioma cells were stereotactically injected into the rat brain to create a tumor. Fifteen days later, MR imaging was used to confirm the presence of a glioma. The animals were randomly assigned to 1 of 3 groups: 1) temozolomide alone (350 mg/m(2)/day for 5 days); 2) temozolomide and radiation therapy (8 Gy); or 3) TSC (100 microg/kg for 5 days), temozolomide, and radiation therapy. Animals were followed through survival studies, and tumor response was assessed on serial MR images obtained at 15-day intervals during a 2-month period. RESULTS: Mean survival (+/- SEM) of the temozolomide-alone and the temozolomide/radiotherapy groups was 23.2 +/- 0.9 and 29.4 +/- 4.4 days, respectively. Mean survival in the TSC/temozolomide/radiotherapy group was 39.8 +/- 6 days, a statistically significant improvement compared with either of the other groups (p < 0.05). Although tumor size was statistically equivalent in all groups at the time of treatment initiation, the addition of TSC to temozolomide and radiotherapy resulted in a statistically significant reduction in the MR imaging-documented mean tumor size at 30 days after tumor implantation. The mean tumor size in the TSC/temozolomide/radiotherapy group was 18.9 +/- 6.6 mm(2) compared with 42.1 +/- 2.7 mm(2) in the temozolomide-alone group (p = 0.047) and 35.8 +/- 5.1 mm(2) in the temozolomide/radiation group (p = 0.004). CONCLUSIONS: In a hypoxic GB model, TSC improves the radiological and clinical effectiveness of temozolomide and radiation therapy. Further investigation of this oxygen diffusion enhancer as a radiosensitizer for hypoxic brain tumors seems warranted. PMID- 20001587 TI - Maintained deep brain stimulation for severe dystonia despite infection by using externalized electrodes and an extracorporeal pulse generator. AB - Infection in the context of implant surgery is a dreaded complication, usually necessitating the removal of all affected hardware. Severe dystonia is a debilitating condition that can present as an emergency and can occasionally be life threatening. The authors present 2 cases of severe dystonia in which deep brain stimulation was maintained despite the presence of infection, using ongoing stimulation by externalization of electrode wires and an extracorporeal pulse generator. This allowed the infection to clear and wounds to heal while maintaining stimulation. This strategy is similar to that used in the management of infected cardiac pacemakers. The authors suggest that this prolonged extracorporeal stimulation should be considered by neurosurgeons in the face of this difficult clinical situation. PMID- 20001589 TI - "In-window" craniotomy and "bridgelike" duraplasty: an alternative to decompressive hemicraniectomy. AB - OBJECT: The object of this study was to propose an alternative procedure to the classic decompressive hemicraniectomy using an "in-window" craniotomy and a "bridgelike" duraplasty. METHODS: The authors performed a large, almost rectangular craniotomy involving the frontal, temporal, and parietal bones and part of the occipital squama in 5 patients. The dura mater is opened and its area is enlarged using a rectangular dural patch of the surgeon's choice in the form of a bridge between the anterior and posterior dural edges. With a vertical cut, the bone flap is divided into 2 similarly sized pieces that function as "window lids." The outer frontal and occipital sides of the bone are tied to the skull border at 2 points to function as a hinge joint. The angle of the bone cut must be beveled outward (inclination ~ 45 degrees of the bone drill or saw) to allow the bone flap to rest on the adjacent skull and prevent its slippage toward the intracranial cavity. RESULTS: The above procedures were performed with effective control of intracranial hypertension due to cerebral venous sinus thrombosis, brain trauma, intracerebral hematoma, or malignant cerebral ischemia. CONCLUSIONS: Decompressive surgery, which uses an in-window craniotomy that gradually opens according to the intracranial pressure, is an alternative solution for deploying autologous material. The procedure has the advantage of obviating the need for a second surgical procedure to close the bone defect, and thus preventing the metabolic cerebral impairment associated with the absence of an overlying skull. PMID- 20001590 TI - The effect of a PP2A inhibitor on the nuclear receptor corepressor pathway in glioma. AB - OBJECT: Nuclear receptor corepressor (N-CoR) forms a complex that maintains neural stem cells in an undifferentiated state through transcriptional repression. Recently, it has been shown that N-CoR is overexpressed in glioblastoma multiforme (GBM) tumor stem cells and has a putative role in maintaining these cells in an undifferentiated immortal state. To determine the effects of disruption of N-CoR complex function by serine/threonine protein phosphatase 2A (PP2A) inhibition on GBM tumor cell differentiation and proliferation, the authors developed and investigated a competitive small molecule inhibitor (LB1) of PP2A in GBM. METHODS: The authors investigated the effects of LB1 on GBM proliferation and molecular differentiation pathways using in vitro and in vivo studies. RESULTS: The LB1 inhibited PP2A, leading to increased levels of phosphorylated Akt kinase and decreased NCoR expression, as well as dose-dependent antiproliferative activity in cultured U87 and U251 malignant glioma cells (dose range 1-10 microM). Systemic LB1 treatment (1.5 mg/kg/day for 21 days) had significant tumor antiproliferative effects in mice harboring U87 glioma xenografts (73% mean reduction in tumor volume compared with controls; p < 0.001). Moreover, a reduction in PP2A expression and activity after LB1 treatment in vivo correlated with increased Akt phosphorylation, reduced nuclear N-CoR expression and N-CoR cytoplasmic translocation, and increased accumulation of acetylated core histones, which coincided with the appearance of glial fibrillary acidic protein-expressing tumor cells. CONCLUSIONS: These findings indicate that PP2A inhibition effectively disrupts N-CoR complex function/expression and leads to cytoplasmic translocation of N-CoR with subsequent tumor cell differentiation and/or death. Therapeutic paradigms that target N-CoR function in the cancer stem cell component of malignant gliomas may have treatment utility. PMID- 20001591 TI - A thermal gel depot for local delivery of paclitaxel to treat experimental brain tumors in rats. AB - OBJECT: Paclitaxel, a cellular proliferation inhibitor/radiation sensitizer, while effective against gliomas in vitro, has poor CNS penetration and dose limiting toxicities when administered systemically. OncoGel (paclitaxel in Re Gel) provides controlled local paclitaxel release when placed into the CNS. The authors evaluated the safety and efficacy of OncoGel in rats with intracranial 9L gliosarcoma. METHODS: Safety studies included intracranial delivery of increasing volumes of ReGel and OncoGel containing 1.5 (OncoGel 1.5) or 6.3 (OncoGel 6.3) mg/ml paclitaxel. An in vivo radiolabeled biodistribution study was performed in 18 Fischer-344 rats to determine intracerebral distribution. Efficacy studies compared overall survival for controls, ReGel only, radiation therapy only, OncoGel 6.3, or OncoGel 6.3 in combination with radiation therapy. ReGel and OncoGel 6.3 were delivered either simultaneously with tumor implantation (Day 0) or 5 days later (Day 5). Radiation therapy was given on Day 5. RESULTS: Control and ReGel animals died of tumor within 17 days. Survival significantly increased in the Onco-Gel 6.3 group on Day 0 (median 31 days; p = 0.0001), in the OncoGel 6.3 group on Day 5 (median 17 days; p = 0.02), and in the radiation therapy-only group (median 26 days; p = 0.0001) compared with controls. Animals receiving both OncoGel and radiation therapy had the longest median survival: 83 days in the group with radiation therapy combined with OncoGel 6.3 on Day 0, and 32 days in the group combined with OncoGel 6.3 on Day 5 (p = 0.0001 vs controls). After 120 days, 37.5% of the animals in the OncoGel Day 0 group, 37.5% of animals in the OncoGel 6.3 Day 0 in combination with radiation therapy group, and 12.5% of the animals in the OncoGel 6.3 on Day 5 in combination with radiation therapy group were alive. In the biodistribution study, measurable radioactivity was observed throughout the ipsilateral hemisphere up to 3 weeks after the OncoGel injection, with the most radioactivity detected 3 hours after injection. The highest dose of radioactivity observed in the contralateral hemisphere was at the Day 3 time point. CONCLUSIONS: OncoGel containing 6.3 mg/ml of paclitaxel is safe for intracranial injection in rats and effective when administered on Day 0. When combined with radiation therapy, the combination was more effective than either therapy alone and should be studied clinically for the treatment of malignant glioma. PMID- 20001593 TI - Superficial siderosis of the central nervous system after ventriculoperitoneal shunt. AB - The authors report a case of superficial siderosis of the CNS that developed after ventriculoperitoneal (VP) shunt placement for normal-pressure hydrocephalus. A 65-year-old woman had undergone VP shunt insertion for normal pressure hydrocephalus. Her gait disturbance, memory disturbance, and urinary incontinence all improved after the procedure. Two years later, however, her gait became ataxic and her appetite became poor. Brain MR imaging revealed a rim of hypointensity on T2-weighted sequences, enveloping the surface of the cortical fissure, cerebellum, and brainstem. Superficial siderosis of the CNS was diagnosed. Steroid administration improved her symptoms. The authors know of only one case of superficial siderosis developing after VP shunt surgery in the English-language literature. Superficial siderosis should be acknowledged as a possible complication of VP shunt. PMID- 20001594 TI - Repeated rupture of a middle meningeal artery aneurysm in moyamoya disease. Case report. AB - A case of moyamoya disease associated with subarachnoid hemorrhage and intracerebral hematoma resulting from repeated rupture of a middle meningeal artery aneurysm is reported. The aneurysm was progressively enlarged over a period of 1 month and was treated with middle meningeal artery embolization. The treatment method is discussed. PMID- 20001596 TI - Indirect cost-effectiveness analyses of abatacept and rituximab in patients with moderate-to-severe rheumatoid arthritis in the United States. AB - OBJECTIVE: To estimate the incremental cost per quality-adjusted life-years (QALYs) for abatacept and rituximab, in combination with methotrexate, relative to methotrexate alone in patients with active rheumatoid arthritis (RA). METHODS: A patient-level simulation model was used to depict the progression of functional disability over the lifetimes of women aged 55-64 years with active RA and inadequate response to a tumor necrosis factor (TNF)-alpha antagonist therapy. Future health-state utilities and medical care costs were based on projected values of the Health Assessment Questionnaire Disability Index (HAQ-DI). Patients were assumed to receive abatacept or rituximab in combination with methotrexate until death or therapy discontinuation due to lack of efficacy or adverse events. HAQ-DI improvement at month 6, after adjustments for control drug (methotrexate) response, was derived from two clinical trials. Costs of medical care and biologic drugs, discounted at 3% annually, were from the perspective of a US third-party payer and expressed in 2007 US dollars. RESULTS: Relative to methotrexate alone, abatacept/methotrexate and rituximab/methotrexate therapies were estimated to yield an average of 1.25 and 1.10 additional QALYs per patient, at mean incremental costs of $58,989 and $60,380, respectively. The incremental cost-utility ratio relative to methotrexate was $47,191 (95% CI $44,810-49,920) per QALY gained for abatacept/methotrexate and $54,891 (95% CI $52,274-58,073) per QALY gained for rituximab/methotrexate. At an acceptability threshold of $50,000 per QALY, the probability of cost effectiveness was 90% for abatacept and 0.0% for rituximab. CONCLUSION: Abatacept was estimated to be more cost effective than rituximab for use in RA from a US third-party payer perspective. However, head-to-head clinical trials and long-term observational data are needed to confirm these findings. PMID- 20001597 TI - Myofibroblasts generated in culture from sclerotic skin lesions of a patient with extensive chronic graft-versus-host disease after allogeneic hematopoietic stem cell transplantation are of recipient origin. AB - Abstract The origin (recipient/donor) of the myofibroblasts mediating fibrosis in sclerodermatous chronic graft-versus-host disease (cGvHD) was investigated. Sclerodermatous specimens obtained from a patient with extensive cGvHD after an HLA-identical sibling bone marrow transplantation were cultured in order to derive tissue myofibroblasts. All proliferating a-SMA+ fibroblastoid cells revealed recipient origin as examined by variable number tandem repeat (VNTR) PCR. This case report shows that fibrosis in sclerodermatous lesions results from the activation and proliferation of locally-derived recipient fibroblasts rather than from donor-derived fibroblasts or circulating fibrocytes. PMID- 20001598 TI - Application of high-resolution skeletal imaging to measurements of volumetric BMD and skeletal microarchitecture in Chinese-American and white women: explanation of a paradox. AB - Asian women have lower rates of hip and forearm fractures despite lower areal BMD (aBMD) by DXA compared with white women and other racial groups. We hypothesized that the lower fracture rates may be explained by more favorable measurements of volumetric BMD (vBMD) and microarchitectural properties, despite lower areal BMD. To address this hypothesis, we used high-resolution pQCT (HRpQCT), a new method that can provide this information noninvasively. We studied 63 premenopausal Chinese-American (n = 31) and white (n = 32) women with DXA and HRpQCT. aBMD by DXA did not differ between groups for the lumbar spine (1.017 +/- 0.108 versus 1.028 +/- 0.152 g/cm(2); p = 0.7), total hip (0.910 +/- 0.093 versus 0.932 +/- 0.134 g/cm(2); p = 0.5), femoral neck (0.788 +/- 0.083 versus 0.809 +/- 0.129 g/cm(2); p = 0.4), or one-third radius (0.691 +/- 0.052 versus 0.708 +/- 0.047 g/cm(2); p = 0.2). HRpQCT at the radius indicated greater trabecular (168 +/- 41 versus 137 +/- 33 mg HA/cm(3); p = <0.01) and cortical (963 +/- 46 versus 915 +/- 42 mg HA/cm(3); p < 0.0001) density; trabecular bone to tissue volume (0.140 +/- 0.034 versus 0.114 +/- 0.028; p = <0.01); trabecular (0.075 +/- 0.013 versus 0.062 +/- 0.009 mm; p < 0.0001) and cortical thickness (0.98 +/- 0.16 versus 0.80 +/- 0.14 mm; p < 0.0001); and lower total bone area (197 +/- 34 versus 232 +/- 33 mm(2); p = <0.001) in the Chinese versus white women and no difference in trabecular number, spacing, or inhomogeneity before adjustment for covariates. Similar results were observed at the weight-bearing tibia. At the radius, adjustment for covariates did not change the direction or significance of differences except for bone, which became similar between the groups. However, at the tibia, adjustment for covariates attenuated differences in cortical BMD and bone area and accentuated differences in trabecular microarchitecture such that Chinese women additionally had higher trabecular number and lower trabecular spacing, as well as inhomogeneity after adjustment. Using the high-resolution technology, the results provide a mechanistic explanation for why Chinese women have fewer hip and forearm fractures than white women. PMID- 20001600 TI - Resilience to bacterial infection: difference between species could be due to proteins in serum. AB - Vertebrates vary in resistance and resilience to infectious diseases, and the mechanisms that regulate the trade-off between these often opposing protective processes are not well understood. Variability in the sensitivity of species to the induction of damaging inflammation in response to equivalent pathogen loads (resilience) complicates the use of animal models that reflect human disease. We found that induction of proinflammatory cytokines from macrophages in response to inflammatory stimuli in vitro is regulated by proteins in the sera of species in inverse proportion to their in vivo resilience to lethal doses of bacterial lipopolysaccharide over a range of 10,000-fold. This finding suggests that proteins in serum rather than intrinsic cellular differences may play a role in regulating variations in resilience to microbe-associated molecular patterns between species. The involvement of circulating proteins as key molecules raises hope that the process might be manipulated to create better animal models and potentially new drug targets. PMID- 20001599 TI - Interindividual variation in functionally adapted trait sets is established during postnatal growth and predictable based on bone robustness. AB - Adults acquire unique sets of morphological and tissue-quality bone traits that are predictable based on robustness and deterministic of strength and fragility. How and when individual trait sets arise during growth has not been established. Longitudinal structural changes of the metacarpal diaphysis were measured for boys and girls from 3 mo to 8 yr of age using hand radiographs obtained from the Bolton-Brush collection. Robustness varied approximately 2-fold among boys and girls, and individual values were established by 2 yr of age, indicating that genetic and environmental factors controlling the relationship between growth in width and growth in length were established early during postnatal growth. Significant negative correlations between robustness and relative cortical area and a significant positive correlation between robustness and a novel measure capturing the efficiency of growth indicated that coordination of the subperiosteal and endocortical surfaces was responsible for this population acquiring a narrow range of trait sets that was predictable based on robustness. Boys and girls with robust diaphyses had proportionally thinner cortices to minimize mass, whereas children with slender diaphyses had proportionally thicker cortices to maximize stiffness. Girls had more slender metacarpals with proportionally thicker cortices compared with boys at all prepubertal ages. Although postnatal growth patterns varied in fundamentally different ways with sex and robustness, the dependence of trait sets on robustness indicated that children sustained variants affecting subperiosteal growth because they shared a common biological factor regulating functional adaptation. Considering the natural variation in acquired trait sets may help identify determinants of fracture risk, because age-related bone loss and gain will affect slender and robust structures differently. PMID- 20001601 TI - Murine responses to endotoxin: another dirty little secret? PMID- 20001602 TI - Attracting predators without falling prey: chemical camouflage protects honeydew producing treehoppers from ant predation. AB - Predaceous ants are dominant organisms on foliage and represent a constant threat to herbivorous insects. The honeydew of sap-feeding hemipterans has been suggested to appease aggressive ants, which then begin tending activities. Here, we manipulated the cuticular chemical profiles of freeze-dried insect prey to show that chemical background matching with the host plant protects Guayaquila xiphias treehoppers against predaceous Camponotus crassus ants, regardless of honeydew supply. Ant predation is increased when treehoppers are transferred to a nonhost plant with which they have low chemical similarity. Palatable moth larvae manipulated to match the chemical background of Guayaquila's host plant attracted lower numbers of predatory ants than unchanged controls. Although aggressive tending ants can protect honeydew-producing hemipterans from natural enemies, they may prey on the trophobionts under shortage of alternative food resources. Thus chemical camouflage in G. xiphias allows the trophobiont to attract predaceous bodyguards at reduced risk of falling prey itself. PMID- 20001603 TI - Hospital-acquired Staphylococcus aureus infections at Texas Children's Hospital, 2001-2007. AB - OBJECTIVE: To document the introduction of the methicillin-resistant Staphylococcus aureus (MRSA) USA300 clone into a children's hospital. Current molecular epidemiology of infections due to the USA300 strain of MRSA in the pediatric healthcare setting remains obscure. DESIGN: Retrospective study of patients with hospital-acquired S. aureus infection during the period from August 1, 2001, through July 31, 2007, at Texas Children's Hospital in Houston. METHODS: Patients with hospital-acquired S. aureus infection from whom an isolate was available for molecular analysis were included. Clinical information was obtained from patient medical records and the electronic hospital information system. S. aureus isolates underwent antimicrobial susceptibility testing, pulsed-field gel electrophoresis, and polymerase chain reaction testing for staphylococcal cassette chromosome (SCC) mec, agr, the diamine N-acetyltransferase gene, and the Panton-Valentine leukocidin genes (pvl). RESULTS: Of 242 patients with hospital acquired S. aureus infection, 147 (61%) had methicillin-susceptible S. aureus infection. Of the 95 MRSA isolates causing hospital-acquired infection, 69 (73%) were USA300 isolates, and that rate did not increase over time. Skin and soft tissue infection (P < .001), onset of infection less than 10 days after admission (P = .007), and lack of comorbidities (P < .001) were associated with hospital acquired MRSA infection caused by the USA300 strain, compared with other isolates (hereafter referred to as non-USA300 isolates). Nine of 10 patients with a S. aureus infection at the time of death were infected with a non-USA300 strain. USA300 carried SCCmec IV, agr I, the diamine N-acetyl transferase gene, and pvl. USA300 isolates were more susceptible to clindamycin, gentamicin, and trimethoprim-sulfamethoxazole than were other non-USA300 isolates (P < .01). CONCLUSIONS: In our patient population, the annual numbers of observed cases of hospital-acquired S. aureus infection have remained constant. USA300 was the most common clone and, compared with other non-USA300 MRSA isolates, was associated with skin and soft tissue infection, early onset of infection after admission, and greater susceptibility to antimicrobial agents. PMID- 20001604 TI - Efficacy of bacteriophage therapy in a model of Burkholderia cenocepacia pulmonary infection. AB - The therapeutic potential of bacteriophages (phages) in a mouse model of acute Burkholderia cenocepacia pulmonary infection was assessed. Phage treatment was administered by either intranasal inhalation or intraperitoneal injection. Bacterial density, macrophage inflammatory protein 2 (MIP-2), and tumor necrosis factor alpha (TNF-alpha) levels were significantly reduced in lungs of mice treated with intraperitoneal phages (P < .05). No significant differences in lung bacterial density or MIP-2 levels were found between untreated mice and mice treated with intranasal phages, intraperitoneal ultraviolet-inactivated phages, or intraperitoneal lambda phage control mice. Mock-infected mice treated with phage showed no significant increase in lung MIP-2 or TNF-alpha levels compared with mock-infected/mock-treated mice. We have demonstrated the efficacy of phage therapy in an acute B. cenocepacia lung infection model. Systemic phage administration was more effective than inhalational administration, suggesting that circulating phages have better access to bacteria in lungs than do topical phages. PMID- 20001605 TI - Tumor doubling time after initial response to transarterial chemoembolization in patients with hepatocellular carcinoma. AB - OBJECTIVE: Following initial transarterial chemoembolization (TACE) of hepatocellular carcinoma (HCC), these tumors can frequently recur, making it important to determine the appropriate follow-up interval after initial response to TACE. We therefore assessed the time taken by new recurrent HCCs to double in volume after an initial response to TACE. MATERIAL AND METHODS: This retrospective cohort study included 73 patients who achieved an initial response after TACE. After intrahepatic distant recurrence of HCC, dynamic CT scans were reviewed by two radiologists without clinical information, and the doubling time of new recurrent nodules was determined. The relationship between doubling time and clinical factors was also analyzed. RESULTS: In 32 of the 73 patients, 40 intrahepatic distant recurrent (IDR) nodules were detected over a median period of 14 months (range 4-46 months). The 1-, 2- and 3-year cumulative IDR rates of patients were 27%, 45% and 65%, respectively. The doubling time of IDR HCC ranged from 20 to 104 days, with a median of 46 days. Over 90% of IDR HCCs had a doubling time of less than 3 months. The doubling time (median 108 days; range 51 129 days) of primary HCC in five patients for whom we had information regarding primary tumors was longer than that of recurrent nodules (median 28 days; range 24-90 days) in the same five patients (P = 0.022). CONCLUSIONS: The median doubling time of IDR HCCs after an initial response was 47 days (range 20-104 days). These findings indicate that the appropriate follow-up interval for dynamic CT in patients with an initial response after TACE is less than 3 months. PMID- 20001606 TI - Urinary diversion after cystectomy for bladder cancer: a population-based study in Sweden. AB - OBJECTIVE: To investigate the type of urinary diversion performed after cystectomy in patients with muscle-invasive bladder cancer in Sweden, using data from a population-based national register. MATERIAL AND METHODS: Since 1997, the Swedish Bladder Cancer Register has included more than 90% of all patients with newly diagnosed bladder cancer. The different types of urinary diversion performed in 1997-2003 were analysed, comparing non-continent diversion (ileal conduit) with continent reconstruction (bladder substitution or continent cutaneous diversion). RESULTS: During the study period, 3463 patients were registered with clinical T2-T4 non-metastatic bladder cancer. Cystectomy was performed in 1141 patients with ileal conduit in 732 (64%) and continent reconstruction in 409 (36%). Ileal conduit was used more frequently in females than males (p = 0.019), in patients older than 75 years (p < 0.00001), and in those with less favourable TNM classification. Continent reconstruction was done more often at university hospitals than at county hospitals (p < 0.00001), but rarely in the northern and western healthcare regions compared with other regions (p < 0.00001). Nationwide, the proportion of registered continent reconstructions decreased, although the absolute number was relatively stable (50-60 per year). CONCLUSIONS: Continent reconstruction after cystectomy for muscle-invasive bladder cancer is performed more often in some healthcare regions and in patients at university hospitals than in county hospitals, indicating a substantial provider influence on the choice of urinary diversion. Over time, the proportion of these procedures has decreased, while the absolute number has remained low and stable; therefore, concentration in high-volume hospitals specialized in bladder cancer and continent reconstruction seems appropriate. PMID- 20001607 TI - Platelet count less than SHARP: what does a case series reveal? AB - Hepatocellular carcinoma (HCC) is increasing in numbers worldwide, and no effective systemic treatment existed for advanced HCC until SHARP (Sorafenib in HCC Assessment Randomized Protocol) study proved sorafenib (Nexavar((R)), Bayer Pharmaceuticals, Wayne, NJ, USA) prolonged survival versus placebo. Child-Pugh class A liver function and a platelet count of > or = 60,000/mm(3) were among the inclusion criteria for SHARP. No safety data in patients with < 60,000/mm(3) of platelets are present. Thrombocytopenia is one of the most frequent challenges faced in patients with chronic liver diseases. We report a series of three patients with HCC and platelet count < 60,000/mm(3) who were successfully treated with sorafenib with no complications. We describe the current data on sorafenib and challenges faced in patients with HCC. In addition, we emphasize the need for informed consent when facing factors that predispose to bleeding (esophageal varices, coagulopathy and thrombocytopenia), possible band ligation before the start of sorafenib, careful clinical monitoring and discontinuation of sorafenib when major bleeding occurs. PMID- 20001609 TI - Validity and reliability of community health activities model program for seniors and short-form international physical activity questionnaire as physical activity assessment tools in patients with fibromyalgia. AB - PURPOSE: There currently is a paucity of well-validated instruments to quantitatively measure physical activity (PA) levels in patients with fibromyalgia syndrome (FMS). This study aims to determine the construct validity and test-retest reliability of two self-report physical activity questionnaires [short-form international physical activity questionnaire (s-IPAQ) and community health activities model program for seniors (CHAMPS)] in a fibromyalgia population. METHODS: Thirty subjects (mean age 49.1 +/- 9.6 years; 90% females) who met the American College of Rheumatology criteria for FMS were invited to participate in the study. Two-week test-retest reliability was evaluated for the CHAMPS and s-IPAQ. Construct validity was evaluated by comparing PA reported from these questionnaires with data obtained from accelerometry (steps/week, counts/week) and the 6-minute walk test (6-MWT). RESULTS: CHAMPS showed greater test-retest reliability (r = 0.58; p < 0.001) compared with s-IPAQ (r = 0.18; p = 0.15). No significant correlations were observed between the self-report PA questionnaires and the 6-MWT or accelerometry data (p > 0.05). CONCLUSIONS: In a fibromyalgia population, the s-IPAQ does not appear to be a reliable and valid PA assessment tool. CHAMPS displayed moderate test-retest reliability; however, no associations were found between CHAMPS and objectives measures of PA. In view of the known benefits of exercise in patients with FMS, there is a need to develop new measures or validate other existing well-established PA questionnaires. PMID- 20001608 TI - Measurement of soluble inflammatory mediators in cerebrospinal fluid of human immunodeficiency virus-positive patients at distinct stages of infection by solid phase protein array. AB - The objective of this study was to evaluate immune cytokine expression in cerebrospinal fluid (CSF) of patients with human immunodeficiency virus-1 (HIV-1) associated dementia (HAD) using a novel cytokine array assay. HIV-1 induces a condition resembling classical subcortical dementia, known as HAD. The immune mechanisms contributing to HAD have not been elucidated. Cytokine expression in CSF was determined by solid-phase protein array in 33 neurologically asymptomatic HIV-positive male patients and were compared to levels in non-HIV controls and patients with HAD. Neurological examinations and lumbar and venous punctures were conducted in all patients and controls. Interleukin (IL)-1, IL-4, and IL-10, were up-regulated in all treated acquired immunodeficiency syndrome (AIDS) patients independent of neurological status compared to controls. In contrast, interferon gamma (IFN-gamma), IL-1alpha, IL-15, and tumor necrosis factor alpha (TNF-alpha) were highly expressed in patients with HAD compared to undemented HIV-positive patients. These results show that solid-phase protein array can detect immunological changes in patients infected with HIV. Cytokine expression levels differ in different disease stages and in patients on different treatment paradigms. Pending further validation on a larger number of patients, this method may be a useful tool in CSF diagnostics and the longitudinal evaluation of patient with HAD. PMID- 20001611 TI - Lamivudine for the treatment of HIV. AB - Lamivudine is a nucleoside reverse transcriptase inhibitor that is widely used for the treatment of HIV-1 infection in combination with other antiretrovirals. It is a highly effective agent that can be dosed once or twice daily due to its long intracellular half-life. It also has one of the best tolerability and long term safety profiles among all antiretroviral agents and continues to be preferred as part of initial or subsequent combination therapy in HIV-infected patients. PMID- 20001610 TI - Efavirenz in the therapy of HIV infection. AB - IMPORTANCE OF THE FIELD: The use of the first generation non-nucleoside reverse transcriptase inhibitor efavirenz (EFV) as a component of first-line antiretroviral therapy has been accepted worldwide. EFV is the only antiretroviral agent currently on the market that has been combined with emtricitabine and tenofovir disoproxil fumarate in a single tablet and administered once daily. AREAS COVERED IN THIS REVIEW: This article reviews efficacy and safety data on EFV and the role of pharmacogenetics in EFV exposure. Published articles and conference presentations on EFV are reviewed. WHAT THE READER WILL GAIN: CYP2B6 genetic polymorphisms influence the metabolism of EFV. The CYP2B6 G to T polymorphism at position 516 is shown to be associated with elevated plasma concentrations and an increase in neurotoxicity of EFV, while the wild-type genotype has been associated with sub-therapeutic concentrations of EFV, potentially leading to the development of viral resistance. This polymorphism is significantly higher in sub-Saharan Africans and African Americans as compared to Hispanic, European and Asian populations. TAKE HOME MESSAGE: The significance of CYP2B6 polymorphism in EFV exposure indicates the need for prospective clinical studies to evaluate the utility of genotype-driven dose adjustments in populations of diverse descent. PMID- 20001612 TI - Ethical issues on preventions and management of blood genetics disorders--Islamic views. AB - Ethical issues are of concern to all members of the health team and the community at large, particularly in relation to chronic and genetic diseases. The fear from the negative social impact, including stigmatization of the carriers of a genetic disease and the affected individual is of a particular concern to the individual and his family. Members of the health team should protect the individual autonomy, observe the ethical principles including beneficence, nonmaleficent, justice, and confidentiality, and maintain high level of professional competence. In addition, Islamic/Arab costumes and traditional issues need to be respected in these communities. This article will outline the prevention and management of common blood genetic disorders in the high of Islamic ethical teachings, where the individuals benefit is paramount. PMID- 20001613 TI - Current understanding in the management of sickle cell disease. AB - Sickle cell disease (SCD), the commonest monogenetic disorder worldwide, represents a major public health burden because of its significant morbidity and mortality. Advances in molecular and cellular biology have resulted in an accumulation of knowledge on sickle cell pathophysiology and broadened our understanding of the complexity of this molecular disease with heterogeneous manifestations. Natural history studies and clinical trials have provided incremental data on clinical features, complications, and predictors of severity in SCD and, above all, have laid important recommendations for prevention and treatment of complications. Disease modifying therapies that have significantly improved survival of SCD patients have been identified over recent years. Despite increasingly successful therapies and better overall survival, patients continue to die especially with increasing age and health providers caring for SCD patients face major challenges. This article will highlight modern management of SCD and its impact on the lives of affected patients. PMID- 20001614 TI - Growth and factors affecting it in thalassemia major. AB - In our thalassemic (T) cohort, 45% of them had height standard deviation score (HtSDS) less than -2 and 56% of them had growth velocity standard deviation score (GVSDS) less than -1. Their mid-arm circumference and triceps skin-fold thickness were decreased versus normal controls. Their circulating insulin-like growth factor-I (IGF-I) concentrations were significantly lower than normal children. Growth hormone (GH) response to provocation with clonidine and glucagon was defective in half of the short T children (peak GH < 7 ng/dL). Some of the short T children, with normal GH response to provocation, had defective spontaneous nocturnal GH secretion. IGF-I generation after one GH injection was reduced in T children than those with GH deficiency (GHD) and constitutional delay of growth and puberty (CDGP). GH therapy for a year significantly increased IGF-I concentrations, GV, and HtSDS in T children but to a lower level compared to those with GHD or CDGP, suggesting partial GH insensitivity. Pubertal induction with human chorionic gonadotropin in T adolescents was associated with increased IGF-I concentrations, GV, and HtSDS. PMID- 20001615 TI - New concept in natural history and management of diabetes mellitus in thalassemia major. AB - Diabetes Mellitus is a major endocrinopathy, which occurs due transfusional haemosiderosis and is found in 20-30% of adult patients with beta-thalassaemia worldwide, accounting for significant morbidity. It is multifactorial with iron loading being the dominant cause and its management poses a clinical challenge. Diabetes in thalassaemia patients is distinct from type 2 diabetes. It is peculiar in many aspects including its pathophysiology and occurs due to insulin resistance as well as islet cell insufficiency. This article reviews the natural history of diabetes in this presentation with emphasis on prevention monitoring and management. Use of MRI techniques may be useful for future monitoring as well as biochemical monitoring to prevent complications of diabetes. Early intervention with intensified chelation may reverse pancreatic function and structural changes as evident from MRI. PMID- 20001616 TI - Current perspectives of fertility and pregnancy in thalassemia. AB - Expectation to have a family is an important aspiration for a better quality of life for the adults with thalassaemia syndrome. Spontaneous fertility can occur in well chelated and transfused patients but majority are infertile due to hypogonadotrophic hypogonadism (HH) and need assisted reproductive techniques (ART). We report our experience of pregnancy following ART in 11 women with ss thalassaemia major, who had HH with functionally intact ovaries. Pregnancy was achieved successfully by ART only after pre-pregnancy assessment in a multidisciplinary setting. Fourteen healthy newborn infants were delivered. Elective caesarean section was performed in 73%. There were 2 sets of twins and one set of triplet pregnancy. No significant cardiac complications were encountered. The incidence of preterm labour and growth restriction were 3 fold higher than the background population. Breast feeding was encouraged in all cases. In conclusion, pregnancy is feasible and safe in women with ss-thalassemia with normal resting cardiac performance and optimised iron overload in specialist centres under a multi disciplinary team. PMID- 20001617 TI - Osteopenia-osteoporosis syndrome in patients with thalassemia: understanding of type of bone disease and response to treatment. AB - With increased life expectancy, thalassaemic bone disease including osteopenia osteoporosis syndrome (OOS) is a major cause of bone pain and fragility fractures especially of the lumbar spine, which may be found in 70-80% adult patients with beta-thalassaemia worldwide, accounting for significant bone morbidity. The causes of OOS in thalassaemia syndromes are multifactorial, and the exact treatment is far from ideal. We undertook a prospective study of 34 thalassaemic patients to evaluate the effect of pamidronate and hormone replacement therapy (if hypogonadal) on bone quantity by DXA scan quality by histomorphometry of bone biopsy and bone dynamics by biochemical markers of bone turnover.Our results show that all patients had osteopenia with abnormal bones on histomorphometry before commencement of treatment. Thalassaemia major patients had high turnover bone disease and all responded favourably to treatment whereas intermedia patients had low turnover bone disease and responded poorly to treatment. Our data also showed that ICTP and PICP is a discriminatory test but histomorphometry is the only test for characterisation of bone morphology. PMID- 20001618 TI - Treatment and prevention of hepatitis B and C in thalassemia. AB - Patients with thalassemia have disturbances in hemoglobin chain production which leads to anemia requiring long-term and multiple transfusions, increasing the risk for transfusion-related viruses, including hepatitis B and C viruses. Although this transfusion-related risk has been virtually eliminated with optimal blood screening practices, a significant number of patients transfused prior to 1990 are infected with hepatitis C virus. Treatment of hepatitis B and hepatitis C has improved rapidly leading to viral eradication - or control of viral replication - with the aim of stopping the progression to liver cirrhosis and its complications. Treatment of chronic hepatitis B in patients with thalassemia does not differ from that of other patients. However, current treatment of hepatitis C necessitates the use of ribavirin, which is associated with dose-dependent hemolysis, requiring adjustment of the transfusion protocol for thalassemia patients during the treatment period. Several measures should be taken in thalassemia patients to prevent viral infection including vaccination for hepatitis B and adequate screening of blood and blood products. PMID- 20001619 TI - Prevention of hemoglobinopathies in Egypt. AB - The hemoglobin disorders are the most common clinically serious single gene disorders in the world. In Egypt, beta-thalassemia is the most common type with a carrier rate varying from 5.3 to > or =9% and a gene frequency of 0.03. So, it was estimated that 1,000/1.5 million per year live births will suffer from thalassemia disease in Egypt (total live births 1,936,205 in 2006). beta Thalassemia creates a social and financial burden for the patients' family and the Egyptian government. The high frequency of beta-thalassemia carriers with increasing rate of newly born cases is a pressing reason for the importance to develop prevention program for beta-thalassemia in Egypt. Sickle-cell disease (SCD) is not frequent in Egypt except in the Oases where the carrier rate varies from 9 to 22%. Our objectives were to provide an in-depth analysis of the current status of hemoglobinopathies in Egypt and what we need for prevention of these diseases. PMID- 20001620 TI - Insight onto the pathophysiology and clinical complications of thalassemia intermedia. AB - Our understanding of the molecular and pathophysiological mechanisms underlying the disease process in patients with thalassemia intermedia (TI) has substantially increased over the past decade. TI encompasses a wide clinical spectrum of beta-thalassemia phenotypes. Some TI patients are asymptomatic until adult life, whereas others are symptomatic from as young as 2 years. A number of clinical complications commonly associated with TI are rarely seen in thalassemia major, including extramedullary hematopoiesis, leg ulcers, gallstones, thrombosis, and pulmonary hypertension. There are a number of options currently available for managing patients with TI, including transfusion therapy, iron chelation therapy, modulation of fetal hemoglobin production, and hematopoietic stem cell transplantation. However, at present, there are no clear guidelines for an orchestrated optimal treatment plan. PMID- 20001621 TI - The hypercoagulable state in thalassemia intermedia. AB - Although the life expectancy of beta-thalassemia patients has markedly improved over the last few years, patients still suffer from many complications of this congenital disease. The presence of a high incidence of thromboembolic events, mainly in beta-thalassemia intermedia, has led to the identification of a hypercoagulable state in these patients. In this review, the molecular and cellular mechanisms leading to hypercoagulability in beta-thalassemia are highlighted, with a special focus on thalassemia intermedia being the group with the highest incidence of thrombotic events as compared to other types of thalassemia. Recommendations for thrombosis prophylaxis in these patients are also discussed. PMID- 20001622 TI - Heart disease in thalassemia intermedia. AB - Thalassemia intermedia (TI) presents with a clinical phenotype ranging between the severe, transfusion-dependent TM and the asymptomatic carrier state. Cardiovascular complications represent the primary cause of mortality both in TM and in TI. Several factors have been reported to interfere in the pathophysiology of cardiovascular abnormalities in TI, including high output state, vascular abnormalities, endothelial dysfunction, elastic tissue disorders, hypercoagulability, valvular abnormalities and, to a less extent, iron overload. These mechanisms end up affecting both left and right heart, hence leading to ventricular remodeling and finally heart failure. PMID- 20001623 TI - Hydroxyurea in the management of thalassemia intermedia. AB - Hydroxyurea (HU) is an antineoplastic agent that enhances fetal hemoglobin. The clinical significance induced by this compound is well known in sickle cell disease. This clinical significance could also be expected in beta-thalassemia patients. Although studies on beta-thalassemia major patients showed significant results, but these clinical responses are expected to be more in thalassemia intermedia (TI) patients because of lesser alpha/beta globin imbalance. Studies showed that HU therapy in TI patients has significant effects on increasing Hb levels that can cause reducing blood transfusion dependency and transfusion free in some patients, decreasing skeletal deformities and splenomegaly and increasing energy state. So HU therapy could be a useful alternative to blood transfusion in some TI patients. PMID- 20001624 TI - Allogeneic stem cell transplantation in patients with beta-thalassemia: King Faisal specialist hospital and research centre experience. AB - From January 1998-July 2006, 62 stem cell transplantation (SCT) were performed on 60 patients with beta-thalassemia from HLA-related match donors. The overall survival (OS) and event free survival (EFS) for all patients were 94 and 77%. The outcome of allogeneic SCT in our experience is satisfactory with OS 92% and EFS 77%. Transplantation at a young age and when the disease is mild offers the best outcome. More advanced disease is associated with higher rate of rejection and severe graft versus host disease. PMID- 20001625 TI - Gene therapy for homozygous beta-thalassemia. Is it a reality? AB - The beta-thalassemias are genetic disorders that are caused by the absent or insufficient production of the beta-chain of hemoglobin. This deficiency causes ineffective erythropoiesis and hemolytic anemia. Without treatment, the severe form of the disease is lethal within the first decade of life. The only curative therapeutic option to date is allogeneic bone marrow transplantation from a matched, related donor, which carries a low risk of morbidity and mortality. Most patients, however, lack a matched donor and are thus managed with palliative therapy, consisting of lifelong transfusion therapy combined with pharmacological chelation to curb iron accumulation. Despite a major improvement in the chelation therapy and supportive care, the major cause of death in these patients is cardiac failure due to secondary hemochromatosis. The goal of globin gene therapy is to offer a potentially curative treatment to patients lacking a matched, related donor, based on the transfer of a regulated beta-globin gene in autologous CD34+ hematopoietic cells collected following G-CSF mobilization. Our clinical trial at Memorial Sloan-Kettering Cancer Center builds on a 20-year long investigation to develop an erythroid-specific vector to regulate beta-globin transgene expression in the progeny of transduced hematopoietic stem cells. To minimize the risks to the patient, the genetically modified cells will be infused after extensive biosafety testing of the transduced cells and following the administration of a reduced intensity (non-myeloablative) conditioning regimen. The protocol will be offered to patients with transfusion-dependent ss thalassemia who are 15 years or older and lack a matched, related donor. PMID- 20001626 TI - Fetal globin induction in beta-thalassemia. AB - Thalassemia patients with persistently high levels of fetal globin typically have less severe anemia, have milder clinical syndromes, and are often transfusion independent. Therefore, the search for molecules exhibiting the property of inducing gamma-globin gene expression and fetal hemoglobin (HbF) production is of great interest. Different pharmacological agents have been studied, namely erythropoietin, short chain fatty acids and cytotoxic agents, azacytidine, and hydroxycarbamide. Hemoglobin F inducers from natural plants, such as angelicin and resveratrol, are powerful inducers of erythroid differentiation and increase HbF in erythroid progenitors of thalassemia patients. Induction of HbF in beta thalassemia patients is expected to be crucial for developing countries unable to sustain the high cost of clinical management of beta-thalassemia patients. PMID- 20001627 TI - Requirements for a reference or expert thalassemia center: the structure/model for centers dealing with chronic/hereditary blood disorders. AB - Chronic disorders, such as the hemoglobin disorders, have a multi-organ involvement and are subject to complications, requiring multidisciplinary care. Most convenient for the patient is the concentration of expertise under one roof and where routine care, such as blood transfusions, can be provided away from acutely ill patients with episodic infections and other conditions. These specialized centers already exist but as yet the standards, which should designate a center as an expert center or a reference center, have not yet been specified or applied. This article examines standards that have been described by two separate authorities in the United States and Europe and suggests the application of these standards to existing or proposed hemoglobinopathy centers. PMID- 20001628 TI - Steps toward the prevention of hemoglobinopathies in the kingdom of Saudi Arabia. AB - Saudi Arabia is known to have high prevalence of hereditary blood disorder similar to neighboring Mediterranean countries, in particular sickle-cell disease and thalassemia. With increasing number of affected individual (Saadallah AA, Rashed MS. J Inherit Metab Dis. 2007;30:482-489), the Ministry of Health (MOH) had implemented a mandatory premarital testing for hemoglobinopathies (sickle cell disease and thalassemia). Since November 2003, all Saudi couples planning for marriage are required to perform the test (Alhamdan NA, Almazrou YY, Alswaidi FM, Coudhry AJ. Genet Med. 2007;9(6):372-377). Besides its preventive efforts, the MOH had also increased its efforts to improve the health care given to affected hereditary blood disease patients. To that effect, a plan to increase the number of specialized hereditary blood disease centers in different regions is existing. In addition, a Consultation Board, Hereditary Blood Disease Advisory Committee, was formed in January 2008. This committee, under the directorate of MOH, took the responsibility of laying down the foundation for internationally high standard of medical care given in treatment center dealing with patients with hereditary blood disorders. In this report, we will shed some light on such accomplishment. PMID- 20001629 TI - Prevention of hemoglobinopathies in Syria. AB - Hemoglobin disorders are highly prevalent in Syria resulting in a significant strain to national resources. To ensure survival and good quality of life of existing patients who are currently almost 8,000 and increasing by almost 800 each year, it is necessary to introduce a national prevention program. This brief report explains the measures that are under discussion currently and proposed for inclusion in a comprehensive control program. PMID- 20001630 TI - Blood transfusion: quality and safety issues in thalassemia, basic requirements and new trends. AB - Guidelines for minimizing risks from alloimmunization, other transfusion reactions, and infection risks are presented based on the Thalassemia International Federation (TIF) guidelines. Future developments including pretreatment of the red cell product that may reduce infection risks are discussed. The rationale for guidelines about the transfusion regime that optimizes the balance between over- and undertransfusing patients is discussed. The optimal approach is still debated and may vary with the patient population in question. Because of changes in the preparation of red cell products in recent years, there is variability in iron content of a 'unit' of blood. It is recommended that each center involved in the treatment of thalassemia knows the average hematocrit and volume of the blood product that they are using, as this impacts on response to chelation therapy. PMID- 20001631 TI - Pathophysiology of transfusional iron overload: contrasting patterns in thalassemia major and sickle cell disease. AB - The pathophysiological consequences of transfusional iron overload largely reflect the pattern of excess iron distribution and include cardiomyopathy, endocrinopathy, cirrhosis, and hepatocellular carcinoma. Since the introduction of desferrioxamine (DFO) in the late 1970s, these complications have fallen substantially but approximately half of the chelated adult patients with thalassemia major (TM) still show evidence of increased myocardial iron loading by MRI. An understanding of the factors that determine the propensity to extrahepatic iron distribution may be a key to minimizing the pathophysiological consequences of transfusional iron overload. Transfused patients with sickle cell disease (SCD) appear less likely to develop these extrahepatic complications, possibly because plasma nontransferrin-bound iron (NTBI) levels are typically lower than in TM patients at matched levels of iron loading. Other mechanisms that may reduce the extrahepatic iron distribution in SCD include raised plasma hepcidin due to chronic inflammation, lower growth differentiation factor 15 (GDF15) levels because of less ineffective erythropoiesis (IE), and induction of heme oxygenase (HO1) by intravascular hemolysis. Further understanding of these mechanisms may help in designing strategies to decrease extrahepatic iron distribution in TM. PMID- 20001632 TI - Iron overload: consequences, assessment, and monitoring. AB - In patients suffering from transfusion-dependent anemia, excess iron secondary to regular transfusions cannot be physiologically excreted. This leads to a state of chronic iron overload with iron accumulating in the liver, heart, and endocrine organs, and ultimately results in significant morbidity and mortality. Historically, iron overload was assessed through measurement of serum ferritin or direct determination of liver iron concentration (LIC) by means of biopsy. Although both correlate well with iron overload severity, several limitations pertinent to both are of concern. This has led to the identification of novel noninvasive iron assessment measures, namely magnetic resonance imaging (MRI) R2 and T2*. Moreover, investigations of other potential indices like nontransferrin bound iron (NTBI) and labile plasma iron (LPI) are yielding promising results. Optimal iron overload assessment and monitoring is a key element in the development of improved strategies of iron chelation therapy that can be tailored to meet the patient's specific needs. PMID- 20001633 TI - Overview of iron chelation therapy with desferrioxamine and deferiprone. AB - Chronic iron overload from frequent blood transfusions to treat patients with severe anemias leads to significant morbidity and mortality. Although desferrioxamine, the current standard of care, is an effective iron chelator with long-term evidence, it requires tedious subcutaneous infusion that reflects negatively on patient compliance. Deferiprone opened the horizon for an era of oral iron chelators. Although collective evidence proved its efficacy, safety issues are still of high concern and require regular monitoring. The experience with these two drugs helps better delineate the optimal goals of iron chelation therapy and the ideal iron chelator. PMID- 20001634 TI - Preimplantation genetic diagnosis. AB - Couples at risk for having an affected child with homozygous thalassemia or other serious hemoglobin disorder have various options for prevention. The most used in some countries has been prenatal diagnosis with a choice of termination of pregnancy. A more recent addition is preimplantation genetic diagnosis (PGD). In this article, this method is described and reviewed. PMID- 20001635 TI - Deferasirox: an update. AB - Cumulative data from prospective studies in over 3,000 patients with a range of transfusion dependent anemias have established key principles about the efficacy and safety of deferasirox (DFS). Pivotal preregistration studies established the relationship between dose, iron excretion, and tolerability using changes in liver iron concentration (LIC) to measure changes in body iron. The importance of the transfusional iron-loading rate to treatment response has been clearly demonstrated. Unwanted effects in these core studies included skin rash and gastrointestinal disturbances. Serum creatinine increments were initially seen in about one-third of patients but have not been progressive in study extensions up to 4.5 years. The proportion of patients with serum ferritin <1,000 microg/L has risen steadily without an increase in adverse events and doses >30 mg/kg/day have been given in >200 patients without an increase in adverse events. The large scale EPIC trial involving >1,600 patients has revealed further insights about the interaction of dose with ferritin trends and safety markers. Cardiac substudies of this trial, in >100 patients with established mild to moderate myocardial iron loading by T2*, show a significant reduction in cardiac iron. PMID- 20001636 TI - Update on survival in thalassemia major. AB - Long-term follow-up of cohorts of patients treated in high-income countries has shown a progressive improvement in life expectancy. Myocardial toxicity from iron overload has been the major cause of mortality; however, there has been a substantial decline in cardiac deaths in recent years, related to switching high risk patients from subcutaneous desferrioxamine to chelation regimes which include the oral chelator deferiprone. The role of deferasirox in enhancing life expectancy is yet to be determined, but it is reasonable to expect an improvement compared with past experience with desferrioxamine. Other causes of mortality will become an increasingly important issue for older thalassemic patients: Surveillance, prophylaxis, and prompt treatment of infection remains essential, and chronic hepatitis virus infection should be managed with best available current therapies. More data on follow-up of thalassemic patients in middle income countries are needed to demonstrate a similar trend in improved survival. The life expectancy for those in low-income countries is similar to the situation 50 years ago in Europe and the United States. The global thalassemia and public health community should consider how to respond to this disparity. PMID- 20001637 TI - Cardiac complications in thalassemia major. AB - Thalassemia major is characterized by chronic ineffective erythropoiesis and anemia as its primary problems. These, in turn, produce physiologic adaptations in the cardiovascular system as well as pathologic/iatrogenic processes such as iron overload, splenectomy, nutritional deficiencies, chronic oxidative stress, and lung disease. This article discusses the pathophysiology of thalassemia as it relates to the cardiovascular system, the mechanisms and monitoring of iron cardiomyopathy, pulmonary hypertension, and vascular aging in thalassemia patients. PMID- 20001638 TI - Assessment and treatment of cardiac iron overload in thalassemia. AB - Cardiac disease remains the major cause of death in thalassemia major and iron overload is involved in heart failure development. Cardiac iron load and heart injury are evaluated by different means, among which CMR measurements remain the best method for estimation. In cases of heart iron overload, chelation treatment should be appropriately tailored in terms of intensification. PMID- 20001639 TI - Sickle cell disease at the dawn of the molecular era. AB - Recent investigations have identified a syndrome of hemolysis-associated vasculopathy in patients with sickle cell disease (SCD), which features severe hemolytic anemia that leads to scavenging of nitric oxide (NO) and its biochemical precursor arginine. This diminished bioavailability of NO promotes several clinical sequelae, which includes pulmonary hypertension, cutaneous leg ulceration, priapism, and ischemic stroke. Additional correlates of this vasculopathy include activation of endothelial cell adhesion molecules and leukocytes, as well as oxidative stress-related pathways. Some known risk factors for atherosclerosis are also associated with sickle cell vasculopathy, including low levels of apolipoprotein AI and high levels of asymmetric dimethylarginine, an endogenous inhibitor of NO synthase. Our current understanding of the dysregulated vascular biology pathways in SCD provides a basis for new clinical trials investigating promising targeted therapeutics. PMID- 20001640 TI - A qualitative study of the influence of poor dental aesthetics on the lives of young adults. AB - OBJECTIVE: Although many countries offer some publicly funded orthodontic treatment for children, not all conditions receive treatment and some adolescents enter adulthood with persisting poor dental aesthetics or malocclusions. The aim of this study was to generate a theory highlighting the main concerns of young adults, either native-born or of immigrant background, with poor dental aesthetics and the measures they adopt to manage their condition in everyday life. MATERIAL AND METHODS: A qualitative method, classic grounded theory, was applied in order to generate a substantive theory highlighting the main concerns and managing mechanisms of 13 strategically selected 19- and 20-year-olds with poor dental aesthetics. Open interviews were conducted with each participant, the topics covering different aspects of social and dental conditions. RESULTS: A core category and three conceptual categories were generated. The core category was labelled "Being under the pressure of social norms" and was related to categories explaining three different ways in which these young adults handle their main concern: (1) avoiding showing their teeth; (2) minimizing the importance of appearance; and (3) seeking orthodontic treatment. The theory offers the potential for improved understanding of young adults who, despite poor dental aesthetics, are managing well with life, and also of those who have not adjusted well. CONCLUSIONS: In early adolescence it may be problematic to make decisions about orthodontic treatment. Undisclosed dental fear can be an important barrier. Some of the young adults in the present study would probably benefit from treatment. PMID- 20001641 TI - Tracking of parents' attitudes to their children's oral health-related behavior Oslo, Norway, 2002-04. AB - OBJECTIVES: To investigate dental beliefs and attitudes of a diverse group of parents from their children when they were aged 3 and 5 years old and to identify possible mediators for a group composed of the parents with the most negative dental attitudes. MATERIAL AND METHODS: Data were collected by parental questionnaire when the children were aged 3 years in 2002 and again 2 years later. The inclusion criteria were children with mothers from Norway (N group) or non-Western countries (IM(1) group). Questionnaires were extensive and had previously been used in a multicenter study. Three composite attitudinal variables relating to oral hygiene, diet and parental indulgence were calculated and an "attitudinal risk group" identified. The association between those variables and the assignment to the group was measured by odds ratio (bivariate and multiple logistic regression). RESULTS: The N parents' dental attitudes were significantly more positive in 2004 when their children were 5 years old than when they were 3 years old (p < 0.0001), but this was not the case among immigrant parents. "Education" and "Immigrant status" [odds ratio (OR) 3.3, 95% confidence interval (CI) 1.6-7.0; and OR 2.8, CI 1.1-7.3, respectively] were significantly associated with the defined "attitudinal risk group". CONCLUSIONS: Only dental attitudes among N parents were significantly more positive in 2004 than in 2002. Not having higher education and being of non-Western background were associated with belonging to the "attitudinal risk group". Culturally tailored programs of dental health education are needed to promote more positive attitudes to oral health. PMID- 20001642 TI - Psychometric evaluation of a new assessment of the ability to manage technology in everyday life. AB - Technology increasingly influences the everyday lives of most people, and the ability to manage technology can be seen as a prerequisite for participation in everyday occupations. However, knowledge of the ability and skills required for management of technology is sparse. This study aimed to validate a new observation-based assessment, the Management of Everyday Technology Assessment (META). The META has been developed to assess the ability to manage technology in everyday life. A sample of 116 older adults with and without cognitive impairment were observed and interviewed by the use of the META when managing their everyday technology at home. The results indicate that the META demonstrates acceptable person response validity and technology goodness-of-fit. Additionally, the META can separate individuals with higher ability from individuals with lower ability to manage everyday technology. The META can be seen as a complement to existing ADL assessment techniques and is planned to be used in both research and practice. PMID- 20001643 TI - Living with rheumatoid arthritis and experiencing everyday life with TNF-alpha blockers. AB - The aim of this study was to describe how persons with RA from an area in western Sweden experience everyday life with TNF-alpha blockers. A purposive sampling of 11 women and four men, with an age ranging from 25 to 70 years, was conducted. A phenomenological approach was used in the study. The data were collected by unstructured in-depth interviews. The data analysis resulted in six code groups, of which four have appurtenant sub-groups. The six code groups are: "A noticeable change dominates the picture"; "Change in bodily and mental symptoms enables activity"; "Enabling care for oneself and others"; "Enabling improved or continued productivity"; "More rewarding leisure time"; and "Drawbacks of the medication". The findings show that most of the informants had experienced dramatic changes in their daily lives since the medication reduced their symptoms, resulting in an increased level of activity. PMID- 20001644 TI - Lessons learned from a multidisciplinary fall-prevention programme: the occupational-therapy element. AB - OBJECTIVE: To gain insight into the contribution of the occupational-therapy part of a multidisciplinary fall prevention programme towards the reduction of falls and functional decline. DESIGN: A descriptive and exploratory study. METHODS: Data were collected in the context of a randomized controlled trial that found no effect of a multidisciplinary fall prevention programme. The study population comprised 166 participants, two occupational therapists (OTs), and one official from each of the five participating municipalities. Data were collected on the recommendations arising from the OT part of the programme, the extent to which those recommendations were implemented and what OTs did to stimulate implementation of behaviour change. RESULTS: The occupational-therapy programme resulted in 457 recommendations; 65% of the recommendations regarding services and assistive devices were implemented. It took on average six months to implement recommended home modifications. Advice on behaviour change predominantly comprised recommendations to reduce risky behaviour. CONCLUSION: To improve the occupational-therapy programme more rapid implementation of recommendations is suggested. Second, participants should be supported to achieve recommended changes. Furthermore, occupational therapists should use theory-based techniques to stimulate behaviour change and use follow-up visits to promote maintenance of the desired behaviour. PMID- 20001645 TI - Reactive nitrogen oxide species induce dilatation of the intercellular space of rat esophagus. AB - OBJECTIVE: Dilatation of the intercellular space (DIS) of the esophageal epithelium is recognized as one of the earliest histological changes in gastroesophageal reflux disease patients. At the human gastroesophageal junction, reactive nitrogen oxide species (RNOS) are generated luminally through the entero salivary re-circulation of dietary nitrate. In cases with gastroesophageal reflux, the site of luminal RNOS generation may shift to the distal esophagus. The aim of this study was to investigate whether luminal RNOS exposure could be involved in the pathogenesis of DIS. MATERIAL AND METHODS: Rat esophageal mucosa was studied with an Ussing chamber model. On the luminal side of the chamber, RNOS were generated by the acidification of physiologic concentrations of sodium nitrite (1.0 or 5.0 mM). Esophageal barrier function was assessed by means of electrophysiological transmembrane resistance and membrane permeability by means of (3)H-mannitol flux. The dimensions of the intercellular spaces were assessed by using transmission electron microscopy. RESULTS: Administration of acid plus sodium nitrite induced DIS of the esophageal epithelium, and this ultrastructural morphological change was accompanied by a concomitant decrease in the transmembrane resistance and an increase in the epithelial permeability. The DIS induced by luminal RNOS was also confirmed in an in vivo exposure model. CONCLUSIONS: The present animal study indicates that the RNOS generated by the acidification of salivary nitrite in the presence of refluxed gastric acid in the esophagus could be a luminal factor that is responsible for the induction of DIS. Further studies are warranted to investigate the clinical relevance of the present findings to the human situation. PMID- 20001646 TI - Acid hypersensitivity in patients with eosinophilic oesophagitis. AB - OBJECTIVE: Painful symptoms are prevalent in patients with eosinophilic oesophagitis but experimental data are sparse. The aim of this study was to compare the pain response to experimental oesophageal stimulation in 14 patients with eosinophilic oesophagitis and 15 healthy volunteers. MATERIAL AND METHODS: A multimodal probe was placed in the oesophagus. The participants were subjected to mechanical, thermal and electrical pain stimuli followed by perfusion with 0.1 M HCl. Pain scores, referred pain areas and evoked brain potentials to electrical stimulation of the oesophagus were recorded. RESULTS: Patients tolerated significantly less acid perfused in the oesophagus (median 123 versus 200 ml; P = 0.02) and felt the burning sensation evoked by the acid earlier (median 2.0 versus 5.0 min; P = 0.01). Eight patients had coexisting gastro-oesophageal reflux disease. Six patients had pure eosinophilic oesophagitis, and this group felt the acid earlier than those with concomitant reflux or the healthy volunteers (median 0.8 versus 2.0 and 5.0 min; P = 0.03). There were no differences between patients and controls in the responses to mechanical or thermal stimulation (P > 0.4). Furthermore, no differences were found for the proxies of central nervous system sensitization (response to electrical stimulations, referred pain areas or evoked brain potentials; P > 0.1). CONCLUSIONS: Patients with eosinophilic oesophagitis are hypersensitive to acid perfused in the oesophagus, and pathophysiologic findings are likely confined to the peripheral tissue. Reflux from physiological acid may play a role in the symptoms of eosinophilic oesophagitis. PMID- 20001648 TI - An EPR method for estimating activity of antioxidants in mouse skin using an anthralin-derived radical model. AB - Inhibitory effects of intravenously or orally administered antioxidants on the anthralin-derived radical generated in skin (mainly in the epidermis) of living mice by ultraviolet-A (UVA) irradiation were estimated. Anthralin was applied to the dorsal skin of living mice and the mice were then exposed to UVA. The EPR signal intensity in skin tissue strips obtained from mice after anthralin-UVA treatment was measured by an X-band EPR spectrometer. Several common antioxidants such as ascorbate, glutathione and Trolox (a vitamin E analogue) intravenously administered to mice reduced anthralin-derived radical generation. Trolox showed the most prolonged and powerful effect. Intravenous injection of a clinically used cerebral neuroprotective drug, Edarabone (Radicut), also showed depletion for the anthralin-derived radical. Oral administration of a commercialized nutritional supplement (a cocktail of 17 herbals and vitamins) also attenuated the anthralin-derived radical. The anthralin-UVA treatment model for antioxidant activity in the epidermis is a potentially feasible method to estimate activity of antioxidants in the body. PMID- 20001647 TI - Traditional reactive carbonyl scavengers do not prevent the carbonylation of brain proteins induced by acute glutathione depletion. AB - This study investigated the effect of reactive carbonyl species (RCS)-trapping agents on the formation of protein carbonyls during depletion of brain glutathione (GSH). To this end, rat brain slices were incubated with the GSH depletor diethyl maleate in the absence or presence of chemically different RCS scavengers (hydralazine, methoxylamine, aminoguanidine, pyridoxamine, carnosine, taurine and z-histidine hydrazide). Despite their strong reactivity towards the most common RCS, none of the scavengers tested, with the exception of hydralazine, prevented protein carbonylation. These findings suggest that the majority of protein-associated carbonyl groups in this oxidative stress paradigm do not derive from stable lipid peroxidation products like malondialdehyde (MDA), acrolein and 4-hydroxynonenal (4-HNE). This conclusion was confirmed by the observation that the amount of MDA-, acrolein- and 4-HNE-protein adducts does not increase upon GSH depletion. Additional studies revealed that the efficacy of hydralazine at preventing carbonylation was due to its ability to reduce oxidative stress, most likely by inhibiting mitochondrial production of superoxide and/or by scavenging lipid free radicals. PMID- 20001650 TI - Undertreatment of hypercholesterolemia. AB - Abstract Recent guidelines recommend strict goals for low-density lipoprotein cholesterol (LDL-C) (1.8-2.6 mmol/L; 70-100 mg/dL). However, these goals are not always met and many primary and secondary prevention patients are not optimally controlled. Both the under-prescription of lipid-lowering medication and lack of adherence to prescribed medications could account for this situation. In this issue of the journal, two studies evaluated the under-treatment of hypercholesterolemia in European countries, as well as patient/physician characteristics that are related to poor control of LDL-C. This editorial considers the implications of these findings. While we have come far in recent years in terms of treating hypercholesterolemia, we still have considerable room for improvement and progress towards evidence-based clinical practice. PMID- 20001649 TI - Defective in vitro IL-2 production in lupus is an early but secondary event paralleling disease activity: evidence from the murine parent-into-F1 model supports staging of IL-2 defects in human lupus. AB - T cell defects are a well described feature of both human and murine lupus however their exact significance is unclear. Evidence from an induced model of lupus, the P --> F1 model of chronic lupus-like GVHD demonstrates that a secondary inducible T cell defect in in vitro IL-2 and CTL responses occurs early in the course of lupus-like disease and well in advance of clinical disease. Defective Th cell function was probed using a novel approach categorizing the response to two stimuli:1) the MHC self restricted response, termed self +X; and 2) the allogeneic response. Using this approach, lupus mice exhibited similar in vitro Th cell pattern i.e. an absent S+X response but preserved allogeneic (termed -/+). In contrast, human lupus patients exhibited three possible response patterns, +/+, - /+ or -/- with more severe in vitro T cell impairment correlated with more severe disease. Similarly, patients with other T cell mediated conditions i.e. HIV infection or renal allograft recipients, also exhibited more severe in vitro T cell impairment with greater disease activity or greater immunosuppression respectively. The similar Th response patterns in human and murine T cell mediated conditions indicates that the underlying mechanisms involved are not disease specific but instead reflect common immune responses and validate the use of the P --> F1 model for future studies of T cell mediated conditions. These results support the use of prospective monitoring of IL-2 responses in lupus patients. Successful adaptation of this approach to the clinical setting could allow not only earlier therapeutic intervention and reduced organ damage but also earlier tapering of pharmacological agents and reduced untoward effects. PMID- 20001651 TI - Predicting short term response to anti-inflammatory therapy in young children with asthma. AB - BACKGROUND: Currently available anti-inflammatory treatment for young children with asthma includes inhaled corticosteroids (ICS) and the leukotriene receptor antagonist (LTRA) montelukast. OBJECTIVE: To evaluate potential biomarkers of predicting short-term (6-week) response to ICS and LTRAs in children with asthma. METHODS: A total of 102 children aged 4 to 7 years with episodic asthma were enrolled in an open labelled single-centre study. Biomarkers and asthma characteristics were evaluated as predictors of treatment. Of 102 patients 45 became symptomatic during observation and were randomised to treatment either to montelukast or fluticasone for 6 weeks. RESULTS: Forced Expiratory Volume in one second (FEV1) increased with both treatments: FEV1 at randomisation was 90.2% and after therapy 106.8% with fluticasone vs. 90.8% and 103.7% for montelukast, respectively, showing that montelukast and fluticasone were equally effective in this age group (p = 0.44). Strong correlations to a favourable treatment response were pre-bronchodilatory FEV1 (p < 0.001) and airway reversibility (p = 0.04) at time of randomisation. None of the other biomarkers (methacholine testing, exhaled nitric oxide [eNO], presence of allergy, total Immunoglobulin E [IgE], cumulative specific IgE, eosinophils and parental smoking) were predictive. CONCLUSION: Despite the small sample size and the open-label design, the study suggests that the use of pre-bronchodilatory FEV1 and airway reversibility appears to be a good indicator of short-term anti-inflammatory therapy in young children with asthma. PMID- 20001652 TI - Should we treat prehypertension? PMID- 20001653 TI - Drug treatment of prehypertension: Not now, not ever? AB - People with prehypertension undoubtedly have an increased risk of cardiovascular and other complications. The vast majority have low absolute risk and whether drug treatment would be beneficial is uncertain. While pharmacotherapy has attractions from a public health prospective, clinicians and crucially those with prehypertension require robust evidence that drug treatment will lead to short term as well as long-term gains. Any changes in recommendations should await adequately powered outcome studies, which provide solid evidence of the magnitude of absolute risk reduction in treating prehyper-tension and assessment of the cost-effectiveness. PMID- 20001654 TI - Reappraisal of European guidelines on hypertension management: a European Society of Hypertension Task Force document. PMID- 20001655 TI - Predictors of cardiovascular events in patients with hypertension and left ventricular hypertrophy: the Losartan Intervention for Endpoint reduction in hypertension study. AB - OBJECTIVE: We assessed readily available patient characteristics, including albuminuria (not included in traditional cardiovascular risk scores), as predictors of cardiovascular events in hypertension with left ventricular hypertrophy (LVH) and developed risk algorithms/scores for outcomes. METHODS: The Losartan Intervention For Endpoint reduction in hypertension (LIFE) study compared effects of losartan-based versus atenolol-based therapy on cardiovascular events in 9193 patients with hypertension and LVH. Univariate and multivariate analyses identified baseline variables with significant impact on development of the primary composite endpoint (cardiovascular death, stroke and myocardial infarction) and its components. Multivariate analysis used a Cox regression model with stepwise selection process. Risk scores were developed from coefficients of risk factors from the multivariate analysis, validated internally using naive and jack-knife procedures, checked for discrimination and calibration, and compared with Framingham coronary heart disease and other risk scores. RESULTS: LIFE risk scores showed increasing endpoint rates with increasing quintile (first to fifth quintile, composite endpoint 2.8-26.7%, cardiovascular death 0.5-14.4%, stroke 1.2-11.3%, myocardial infarction 1.4-8.1%) and were confirmed with a jack-knife approach that adjusts for potentially optimistic bias. The Framingham coronary heart disease and other risk scores overestimated risk in lower risk patients and underestimated risk in higher risk patients, except for myocardial infarction. CONCLUSION: A number of patient characteristics predicted cardiovascular events in patients with hypertension and LVH. Risk scores developed from these patient characteristics, including albuminuria, strongly predicted outcomes and may improve risk assessment of patients with hypertension and LVH and planning of clinical trials. PMID- 20001656 TI - Bayesian approach to bipolar guidelines. PMID- 20001657 TI - Duration of untreated illness as a predictor of treatment response and remission in obsessive-compulsive disorder. AB - OBJECTIVES: The Duration of Untreated Illness (DUI), defined as the time elapsing between the onset of a disorder and the beginning of the first pharmacological treatment, has been increasingly investigated as a predictor of outcome and course across different psychiatric disorders. Purpose of this naturalistic study was to evaluate the influence of DUI on treatment response and remission in a sample of patients with obsessive-compulsive disorder (OCD). METHODS: Sixty-six outpatients with a DSM-IV diagnosis of OCD were included in the study. Patients received, according to their clinical conditions, an open pharmacological treatment of 12 weeks and were evaluated by the administration of the Yale Brown Obsessive Compulsive Scale (Y-BOCS) at baseline and endpoint. Treatment response was defined as a decrease .25% on Y-BOCS score compared to baseline, while remission was defined as an endpoint Y-BOCS total score #10. A logistic regression was performed considering DUI as the independent continuous variable and treatment response and remission as the dependent variables. Moreover, the sample was divided into two groups according to a categorical cut-off for the DUI of 24 months and odds ratios (OR) were calculated on the basis of the same variables. RESULTS: DUI, considered as a continuous variable, was not predictive of treatment response (OR51.00, P50.15) nor remission (OR51.00, P50.59). When considered as a categorical variable, however, a DUI # 24 months was predictive of treatment response (OR50.27, P50.03). CONCLUSIONS: Results from the present naturalistic study suggest a complicated relationship between DUI and treatment outcome in OCD encouraging further investigation with larger samples in order to better define long versus short DUI in this condition. PMID- 20001658 TI - Centralized Pan-European survey on the under-treatment of hypercholesterolaemia (CEPHEUS): overall findings from eight countries. AB - BACKGROUND: Surveys evaluating plasma lipid goal attainment in patients with coronary heart disease have shown that hypercholesterolaemia is inadequately treated. Limited data account for the reasons behind this. The aim of the CEntralized Pan-European survey on tHE Under-treatment of hypercholeSterolaemia (CEPHEUS) survey was to evaluate the current use and efficacy of lipid-lowering drugs (LLD), and to identify possible patient/physician characteristics associated with failure to achieve low-density lipoprotein cholesterol (LDL-C) targets recommended by the 2003 European guidelines (Third Joint Task Force). METHODS: CEPHEUS was a European, multi-centre, cross-sectional survey conducted in eight countries and involved patients on LLD for >3 months (stable medication >6 weeks). One visit was scheduled for data collection, including fasting lipids. In all but one country, physicians and patients filled in a questionnaire about aspects of hypercholesterolaemia and treatment. RESULTS: Of the 15 199 patients recruited, 14 478 were included in the final analyses. The mean age was 63.2 years, and 45% of patients were female. Overall, 55.3% of the patients achieved their LDL-C target. In multivariate analyses, the factors identified as positive predictors for achieving LDL-C goals included normal body mass index, not smoking, not having metabolic syndrome, being on statin therapy and good treatment adherence. LIMITATIONS: The population was a selected group of subjects treated with LLD, and the results cannot be extrapolated to the general population. Patient consent was obtained, which may have selected more motivated patients and induced a positive bias. The physician and patient questionnaires were not validated, but were only used for exploratory purposes. CONCLUSION: Only 55.3% of patients using LLD achieved the LDL-C target recommended in the 2003 European guidelines. PMID- 20001659 TI - Development and validation of a new instrument to evaluate the ease of use of patient-controlled analgesic modalities for postoperative patients. AB - OBJECTIVE: To describe the development and psychometric evaluation of a questionnaire assessing the ease of use that patients associate with patient controlled analgesia (PCA) modalities. METHODS: Qualitative interviews were conducted with patients who had experience with intravenous (IV) PCA for postoperative pain management to generate items relevant to the ease of using PCA modalities. The content validity of the resulting questionnaire was examined through follow-up patient interviews, and an expert panel reviewed the questionnaire. Cognitive debriefing interviews were conducted with patients to determine the clarity and content of the instructions, items, and response scales, and the ease of completing the instrument. Psychometric evaluation was performed with patients who had undergone surgery and received IV PCA for postoperative pain management. Item and scale quality and the internal consistency reliability of the questionnaire were assessed. Construct validity was evaluated by examining the relationship between subscales of the questionnaire with patient-reported outcome measures. Known-groups validity was determined by assessing the instrument's ability to differentiate between patients with versus without an IV PCA problem. A potential limitation of this study was the exclusive sampling of patients who had experience with IV PCA. RESULTS: The Patient Ease-of-Care (EOC) Questionnaire included 23 items in the following subscales: Confidence with Device, Comfort with Device, Movement, Dosing Confidence, Pain Control, Knowledge/Understanding, and Satisfaction. Coefficient alpha reliability estimates were >= 0.66 for Overall EOC (includes all subscales except Satisfaction) and all EOC subscales. Construct validity was supported by the moderate relationship between the Pain Control subscale and measures of pain severity and pain interference; additional evidence of construct validity was provided by correlations of the Confidence with Device subscale, the Satisfaction subscale, and Overall EOC with measures of pain severity, pain interference, and satisfaction. Significant mean score differences were reported between participants with and without IV PCA problems for Overall EOC and for the Comfort with Device, Confidence with Device, Movement, Pain Control, and Satisfaction subscales indicating known-groups validity. CONCLUSIONS: Results provide evidence for the reliability and validity of the Patient EOC Questionnaire as a measure of the ease of use that patients associate with PCA systems and may be useful for evaluating emerging PCA modalities. PMID- 20001660 TI - The potential dermal irritating effect of residual (meth)acrylic monomers in pressure sensitive adhesive tapes. AB - It is generally thought that residual unpolymerized (meth)acrylic monomers commonly found in pressure sensitive adhesive tapes for medical use may cause dermal irritation, but a systematic study has never been carried out. Therefore, we assessed the potential dermal irritating effect of residual (meth)acrylic monomers. We studied seven acrylic monomers, acrylic acid (AA), methyl acrylate (MA), ethyl acrylate (EA), n-butyl acrylate (n-BA), n-hexyl acrylate (n-HA), 2 ethylhexyl acrylate (2-EHA) and 2-hydroxyethyl acrylate (HEA), as well as three methacrylic monomers, methacrylic acid (MAA), methyl methacrylate (MMA) and 2 hydroxyethyl methacrylate (2-HEMA). We first examined their cytotoxic effect on a cultured dermis model using the MTT method to determine their EC(50) and then performed a primary irritation test in rabbits using the monomers at three different concentrations (i.e., EC(50) , one-tenth EC(50) and 10 times EC(50)). Marked variations were found in cytotoxic and dermal irritating activities among the (meth)acrylic monomers tested. HEA exhibited the most potent dermal irritation having the lowest erythema dose (the concentration which gives a primary dermal irritation index of 1.00) of 460 ppm. But the other monomers exhibited less potent dermal irritation (lowest erythema doses > or =1000 ppm). For the monomers, significant correlation was found between cytotoxic activity and in vivo dermal irritating activity. Our results show that residual unpolymerized (meth)acrylic monomers in adhesive tapes are unlikely to induce skin irritation except for HEA. This study also suggests that cultured skin models are extremely useful as a screening method for chemical substances that could potentially cause dermal irritating activity. PMID- 20001661 TI - The protective effect of beta-1,3-D-glucan on taxol-induced hepatotoxicity: a histopathological and stereological study. AB - The present study was undertaken to determine the histopathological and quantitative effects of the antineoplastic agent, taxol, on the liver. The protective effects of the strong antioxidant, beta-1,3-D-glucan, against liver damage induced by taxol were also investigated. Mice were divided into four main treatment groups: control, taxol, beta-1,3-D-glucan, and taxol+beta-1,3-D-glucan. Each group was further subdivided into six subgroups, according to time of sacrifice (6, 12, 24, and 48 hours and 7 and 14 days). After the experiments, quantitative and histopathological changes in liver were examined by light microscopy and modern stereological systems. Stereological results indicated that the portal triad area of the taxol group was significantly reduced, compared to the controls at 12 hours, whereas in the taxol plus beta-glucan and beta-glucan groups, the means were similar to those of the controls. There was no statistically significant difference in the numerical density of hepatocytes with time between the control and other groups. The histopathological results indicated an increased, time-dependent degeneration and necrosis of the liver tissues in mice in the taxol group. Regenerative changes in livers of mice in the taxol plus beta-glucan group were observed, when compared with those of the taxol group. Stereological and histopathological results suggest that beta-glucan may reduce taxol-induced hepatic damage by blocking the change in the portal area and suppressing processes leading to necrosis. PMID- 20001662 TI - General pharmacological properties, developmental toxicity, and analgesic activity of gambogic acid, a novel natural anticancer agent. AB - In this article, the general pharmacological toxicity of gambogic acid (GA), a new anticancer agent, on the dog cardiovascular and respiratory system and the mouse central nervous system (CNS) were observed. The developmental toxicity and analgesic activities of GA were also investigated in rats and mice. Results showed that GA did not cause any toxic symptoms on blood pressure (i.e., mean arterial pressure), heart rate (HR), and respiratory frequency. However, a high dose of GA showed slight side effects on the mouse CNS. Further, evidence of maternal and developmental toxicity was observed in a dose-dependent manner. The maternal body-weight gain, as well as the birth weights and live birth index, were decreased significantly in the treatment groups. The inhibitory effects of GA on fetal skeletal development were also found. No obvious effects of GA on external alterations and visceral alterations were shown. In the analgesic experiments, GA showed significant analgesic activity in the acetic-acid-induced writhing study in a dose-dependent manner. This mechanism might be related to its anti-inflammation properties. PMID- 20001663 TI - Chromosomal damage induced by vanadium oxides in human peripheral lymphocytes. AB - Fly ash, the inorganic residue resulting from the combustion of some fuels, may almost exclusively contain vanadium oxides, compounds which exert potential toxic effects on a wide variety of in vitro and in vivo biological systems. Because information related to the oxidation state responsible for inducing genotoxic effects is controversial, the aim of the present study was to evaluate the effects of three vanadium salts in vitro. Human peripheral lymphocyte cultures were exposed to 1, 2, 4, or 8 microg/mL of vanadium(III) trioxide, vanadium(IV) tetraoxide, or vanadium(V) pentoxide (V(2)O(3), V(2)O(4), or V(2)O(5), respectively). These cultures were then screened for structural chromosomal aberrations, and mitotic index (MI) measurements were made. Cytogenetic evaluations showed that only V(2)O(4) increased the percentage of aberrant cells (without gaps) and chromosome damage (including and excluding gaps), while all compounds led to a decrease in the MI. These results demonstrate that vanadium(III), vanadium(IV), and vanadium(V) are all capable of inducing cytotoxicity, but only oxidation state IV induces clastogenic effects. PMID- 20001664 TI - Evaluation of genotoxicity of Yanhuanglian dehydrocavidine (YHL-DC) in vitro and in vivo. AB - It is reported that dehydrocavidine (DC), the main component of a traditional Chinese medicine, Yanhuanglian (YHL), can protect hepatic tissue against HBV and HAV impairment. As part of a safety evaluation on YHL-DC for use in the treatment of HBV, the present study evaluated the potential genotoxicity of YHL-DC by using the standard battery of tests (i.e., bacterial reverse mutation, chromosomal aberrations, and mouse micronucleus assays) recommended by the State Food and Drug Administration of China. The results showed that YHL-DC was not genotoxic under the conditions of the reverse mutation, chromosomal aberrations, and mouse micronucleus assay conditions. The anticipated clinical dose should be smaller than the doses used in the genotoxicity assays. With confirmation from further toxicity studies, YHL-DC would hopefully prove to be a useful anti-HBV agent. PMID- 20001666 TI - Spleen tyrosine kinase: an Src family of non-receptor kinase has multiple functions and represents a valuable therapeutic target in the treatment of autoimmune and inflammatory diseases. AB - Spleen tyrosine kinase (Syk) is involved in the development and function of B and T cells, the Fc receptor-mediated degranulation of basophils and mast cells. Recent work has assigned important roles for Syk in the aberrant function of T cells in patients with systemic lupus erythematosus (SLE), osteoclasts, and urate crystal-induced neutrophil stimulation. Preclinical and early clinical studies have urged Syk inhibition for the treatment of patients with rheumatoid arthritis, whereas ex vivo experiments and preclinical studies point to a therapeutic potential of Syk inhibition in patients with SLE and crystal-induced arthritides. PMID- 20001672 TI - CYP3A catalyses schizandrin biotransformation in human, minipig and rat liver microsomes. AB - Schizandrin is recognized as the major absorbed effective constituent of Fructus schisandrae, which is extensively applied in Chinese medicinal formula. The present study aimed to profile the phase I metabolites of schizandrin and identify the cytochrome P450 (CYP) isoforms involved. After schizandrin was incubated with human liver microsomes, three metabolites were isolated by high performance liquid chromatography (HPLC) and their structures were identified to be 8(R)-hydroxyl-schizandrin, 2-demethyl-8(R)-hydroxyl-schizandrin, 3-demethyl 8(R)-hydroxyl-schizandrin, by liquid chromatography-mass spectrometry (LC-MS), (1)H-nuclear magnetic resonance (NMR), and (13)C-NMR, respectively. A combination of correlation analysis, chemical inhibition studies, assays with recombinant CYPs, and enzyme kinetics indicated that CYP3A4 was the main hepatic isoform that cleared schizandrin. Rat and minipig liver microsomes were included when evaluating species differences, and the results showed little difference among the species. In conclusion, CYP3A4 plays a major role in the biotransformation of schizandrin in human liver microsomes. Minipig and rat could be surrogate models for man in schizandrin pharmacokinetic studies. Better knowledge of schizandrin's metabolic pathway could provide the vital information for understanding the pharmacokinetic behaviours of schizandrin contained in Chinese medicinal formula. PMID- 20001673 TI - In vitro mammalian metabolism of the mitosis inhibitor zoxamide and the relationship to its in vitro toxicity. AB - The in vitro mammalian metabolism of the fungicide zoxamide is related to its in vitro mammalian toxicity. After incubation of zoxamide with rat liver microsomes leading to practically 100% metabolism (mostly hydroxylated zoxamide), the cytotoxicity (methyl thiazole tetrazolium (MTT) test) and the mitosis-inhibiting potential (shown by cell count and by cell cycle analysis) for V79 were not distinguishable from those of zoxamide, demonstrating that the hydroxylation of zoxamide did not change the cytotoxicity or mitosis-inhibiting potential as determined by these assays. After incubation of zoxamide with rat liver S9 predominantly leading to conjugation with glutathione, and after incubation of zoxamide with rat liver slices predominantly leading to the glucuronide of the hydroxylated zoxamide, these activities were eliminated demonstrating that the glutathione conjugate and the glucuronide had lost the activities in these assays due either to no intrinsic potential of these conjugates or to their inability to penetrate the plasma membrane of mammalian cells. It is concluded that the metabolic hydroxylation of zoxamide did not change its activity in the assays used for investigating its influence on cell proliferation, cell cycle and cytotoxicity, while the formation of conjugates with glutathione or glucuronic acid led to the apparent loss of these activities. Thus, with zoxamide as a prototype, it was shown that, in principle, mammalian metabolism and its relationship to mammalian detoxication of fungicidal mitosis inhibitors may be reasonably anticipated from in vitro studies. In addition, the results provide a rational for the observed absence of typically mitosis inhibition-associated toxicities of zoxamide in mammals in vivo. PMID- 20001675 TI - New features and performance of a next-generation SEVEN-day continuous glucose monitoring system with short lag time. AB - BACKGROUND: The purpose of this study was to evaluate new features and performance of the SEVEN PLUS System (DexCom, Inc., San Diego, CA), a real-time continuous glucose monitoring (CGM) device. This study is the first to evaluate the SEVEN PLUS device. METHODS: Fifty-three subjects were enrolled at three U.S. centers; 43 (81%) of the subjects had type 1 diabetes mellitus, and 10 (19%) had insulin-requiring type 2 diabetes mellitus. Subjects inserted and wore either one or two sensors for 7 days. A subgroup (n = 18) wore two sensors to track precision. Subjects participated in one 8-h in-clinic session with blood draws every 15 min on study Day 1, 4, or 7 to collect laboratory reference YSI instrument (YSI, Yellow Springs, OH) and self-measured plasma glucose (SMPG) reference measurements. For the remainder of the week, CGM was used as an adjunct to SMPG during home use. RESULTS: The overall median absolute relative difference (ARD) versus YSI was 13.0%. Zones A and B of the Clarke Error Grid of CGM measurements were 73.8% and 22.1%, respectively. Precision ARD was 15.3 +/- 6.2% (mean +/- SD). The median ARD versus SMPG was 12.1%. Sensor life (89% lasted 7 days) was improved compared to the SEVEN device. The lag time obtained with various statistical measures between CGM and YSI were similar and estimated as a median of 8 min (interquartile range, 11 min) using the Pearson correlation coefficient. No serious adverse events or infectious complications were reported. CONCLUSIONS: The performance of this new system compares favorably to the previous SEVEN device in terms of accuracy, precision, lag time, sensor life, and rate of data capture. PMID- 20001676 TI - Responses to continuous glucose monitoring in subjects with type 1 diabetes using continuous subcutaneous insulin infusion or multiple daily injections. AB - OBJECTIVE: We compared changes in response to unmasking of continuous glucose monitoring (CGM) in subjects with type 1 diabetes who use multiple daily injections (MDI) or continuous subcutaneous insulin infusion (CSII). RESEARCH DESIGN AND METHODS: Use of real-time CGM (DexCom [San Diego, CA] SEVEN was studied in 38 subjects using CSII and 26 using MDI. CGM output was masked during Week 1 and unmasked during Weeks 2 and 3. We evaluated changes in 16 criteria for quality of glycemic control and eight criteria for glycemic variability. RESULTS: All 24 criteria showed highly statistically significant improvement when considered simultaneously (P < 0.000001). For subjects using CSII, 18 of 24 criteria improved significantly (nominal P < 0.05); for subjects using MDI, 16 of 24 criteria improved significantly (P < 0.05). Twelve of the comparisons remained significant (P < 0.05) after applying the overconservative Bonferroni correction for multiple comparisons. The percentage of glucose values within the range 80 140 mg/dL increased by 19% and 17% relative to their control values (Week 1) for subjects using MDI and CSII, respectively. Mean glucose, overall SD (SD(T)), SD between daily means (SD(dm)), mean amplitude of glycemic excursion (MAGE), and mean of daily differences (MODD) improved significantly. Responses to CGM display were not significantly different between the MDI and CSII subject groups for any of the 24 criteria considered individually or in groups of eight, 16, or 24. CONCLUSION: CGM has similar effectiveness in subjects with type 1 diabetes using either CSII or MDI. PMID- 20001674 TI - Neuronal and axonal degeneration in experimental spinal cord injury: in vivo proton magnetic resonance spectroscopy and histology. AB - Longitudinal in vivo proton magnetic resonance spectroscopy (1H-MRS) and immunohistochemistry were performed to investigate the tissue degeneration in traumatically injured rat spinal cord rostral and caudal to the lesion epicenter. On 1H-MRS significant decreases in N-acetyl aspartate (NAA) and total creatine (Cr) levels in the rostral, epicenter, and caudal segments were observed by 14 days, and levels remained depressed up to 56 days post-injury (PI). In contrast, the total choline (Cho) levels increased significantly in all three segments by 14 days PI, but recovered in the epicenter and caudal, but not the rostral region, at 56 days PI. Immunohistochemistry demonstrated neuronal cell death in the gray matter, and reactive astrocytes and axonal degeneration in the dorsal, lateral, and ventral white-matter columns. These results suggest delayed tissue degeneration in regions both rostrally and caudally from the epicenter in the injured spinal cord tissue. A rostral-caudal asymmetry in tissue recovery was seen both on MRI-observed hyperintense lesion volume and the Cho, but not NAA and Cr, levels at 56 days PI. These studies suggest that dynamic metabolic changes take place in regions away from the epicenter in injured spinal cord. PMID- 20001677 TI - Difference in insulin usage patterns with pubertal development in children with type 1 diabetes during transition from multiple daily injections to continuous subcutaneous insulin infusion (CSII) and through the CSII treatment. AB - OBJECTIVE: This study analyzed the changes in insulin requirement in the transition from multiple daily injections (MDI) to continuous subcutaneous insulin infusion (CSII) and the differences through the CSII treatment in pediatric patients in different pubertal developmental stages. METHODS: We analyzed, through a longitudinal retrospective study, the insulin usage patterns and glycemic control of 40 patients with type 1 diabetes on CSII treatment for 12 months. The patients were subdivided in three groups: group A, 13 prepubertal subjects (Tanner stage I); group B, 15 pubertal subjects (Tanner stage II-IV); and group C, 12 postpubertal subjects (Tanner stage V). RESULTS: During the transition from MDI to CSII, the insulin requirements decreased significantly by 21 +/- 5% (0.89 +/- 0.26 U/kg/day vs. 0.70 +/- 0.11 U/kg/day). Through the CSII treatment the percentage of total daily insulin delivered as the basal rate in groups A, B, and C was 54-60%, 52-54%, and 52-54%, respectively. The number of basal rates per day was significantly higher in groups A and B. The overall profile of basal rate differed among the groups. During the night, prepubertal patients required more insulin from 12 p.m. to 2 a.m., whereas pubertal and postpubertal patients required more insulin between 2-3 a.m. to 7 a.m. Prepubertal patients had a maximum basal rate between 9 a.m. to 12 a.m. and 2 p.m. to 4 p.m., in contrast to pubertal and postpubertal patients between 2 p.m. to 3 p.m. and 5 p.m. to 6 p.m. The number of "extra" boluses per day was significantly higher in groups B and C compared to group A. CONCLUSIONS: Age related differences exist in insulin usage patterns of pediatric patients on pump treatment. A greater knowledge of these variations may help to obtain optimum conditions in the CSII treatment in pediatric patients. PMID- 20001678 TI - Meta-analysis of the benefits of self-monitoring of blood glucose on glycemic control in type 2 diabetes patients: an update. AB - BACKGROUND: Our systematic review and meta-analysis of the benefit of self monitoring of blood glucose (SMBG) in improving glycemic control in type 2 diabetes was published in 2008. With the few studies that have emerged afterward, we undertook subsequent meta-analysis of the available evidence to update the results. METHODS: Clinical trials of SMBG were identified through electronic searches (MEDLINE, EMBASE, and The Cochrane Library) up to and including June 2009. Studies were included if they met the following inclusion criteria: (1) randomized controlled trial comparing SMBG versus non-SMBG in type 2 diabetes patients not using insulin and (2) hemoglobin A1c (HbA(1c)) reported as an outcome measure. The efficacy was estimated with the mean difference in the changes of HbA(1c) from baseline to final assessment between the SMBG and the non SMBG groups. RESULTS: SMBG was effective in reducing HbA(1c) in non-insulin treated type 2 diabetes (pooled mean difference, -0.24%; 95% confidence interval, -0.34% to -0.14%; P < 0.00001). Glycemic control significantly improved among the subgroup of patients whose baseline HbA(1c) was >or=8%. In contrast, no significant effect of SMBG was detected in patients who had HbA(1c) <8%. CONCLUSIONS: The available evidence suggests the usefulness of SMBG in improving glycemic control in non-insulin-treated type 2 diabetes as demonstrated by the reduction of HbA(1c) levels. In particular, SMBG proved to be useful in the subgroup of patients whose baseline HbA(1c) was >or=8%. PMID- 20001679 TI - Ileal interposition with sleeve gastrectomy for control of type 2 diabetes. AB - BACKGROUND: Bariatric surgery offers the best solution in management of obesity and related metabolic ailments, paving the way for a concept termed metabolic surgery. We report the results of a novel surgical procedure on glycemic control and metabolic syndrome in poorly controlled type 2 diabetes. METHODS: Ten patients (four men, six women) underwent laparoscopic surgical procedure of sleeve gastrectomy and ileal interposition. All patients had diabetes for more than 3 years with poor control despite use of oral hypoglycemic agents (OHAs) and/or insulin. The primary outcome was remission of diabetes (hemoglobin A1c <7% without OHAs/insulin), and secondary outcomes were change in OHA requirement, components of metabolic syndrome, insulin resistance, and microalbuminuria. RESULTS: We report the preliminary postoperative follow-up data of 9.1 +/- 5.3 months (range, 2-16 months). Participants had a mean age of 48.2 +/- 9 years (range, 34-62 years), duration of diabetes of 11 +/- 5.7 years (range, 4-25 years), and preoperative body mass index of 33.8 +/- 6.5 kg/m(2). Seven patients had diabetes remission, and the remaining three showed significantly decreased OHA requirement. All participants had weight loss ranging between 15% and 30% and had remission of hypertension. Microalbuminuria (96.8 +/- 19.1 vs. 46.7 +/- 10.1 mg/L, P = 0.03568) and insulin resistance as assessed by homeostasis assessment model of insulin resistance (5.2 +/- 2.1 vs. 1.8 +/- 0.9, P = 0.0005) decreased significantly after surgery. CONCLUSIONS: Our preliminary observations demonstrated the feasibility, safety, and efficacy of this novel surgical procedure in type 2 diabetes. Further long-term data from more patients are necessary to confirm these findings. PMID- 20001680 TI - The influence of environmental temperature on the response of the skin to local pressure: the impact of aging and diabetes. AB - BACKGROUND: To protect against ischemia, pressure-induced vasodilation (PIV) causes an increase in skin blood flow. Endothelial dysfunction, which is commonly found in older patients and those with diabetes, and global temperatures can affect the resting blood flow in skin, which may reduce the blood flow during and after the application of local pressure. The present study investigated the PIV of the skin with exposure to three global temperatures in younger and older populations and those with diabetes. MATERIALS AND METHODS: Older subjects (n = 15, mean age 64.2 +/- 14.0 years), subjects with diabetes (n = 15, mean age 62 +/ 5.9 years, mean duration 13.2 +/- 9.1 years), or younger subjects (n = 15, mean age 25.7 +/- 2.9 years) participated. An infrared laser Dopler flow meter was used to measure skin blood flow on the bottom of the foot, lower back, and hand during and after applications of pressure at 7.5, 15, 30, 45, and 60 kPa at 16 degrees C, 24 degrees C, and 32 degrees C global temperatures. RESULTS: The resting blood flow for all subjects was significantly lower in the 16 degrees C environment (P < 0.05). Blood flow in the group with diabetes was significantly lower at rest, during the application of all pressure, and after the release of pressure in all global temperatures (P < 0.05). The younger group showed a significant increase in blood flow after every pressure application, except 7.5 kPa, in all global conditions (P < 0.001). Older subjects and patients with diabetes did not have a significant reactive hyperemia, especially in the 16 degrees C environment. CONCLUSION: The protective mechanism of PIV is severely reduced in older populations and those with diabetes, especially in colder environments where skin blood flow is already diminished. PMID- 20001681 TI - Prevalence of pancreatic diabetes in patients carrying mutations or polymorphisms of the PRSS1 gene in the Han population. AB - OBJECTIVE: This study updated the estimated prevalence of type 3c diabetes damage to the pancreas through different genotypes of PRSS1 and their clinical characteristics in the Han population. SUBJECTS AND METHODS: Cross-sectional analysis was performed of the most recent (2003-2007) patients with pancreatitis from six hospitals of the Han population in South China (n = 253). RESULTS: There were 32 patients with pancreatitis carrying a PRSS1 gene abnormality within intron region among 253 cases of pancreatitis, including 27 patients carrying novel single nucleotide polymorphisms, namely, IVS 3 +75 A --> G conversion, and five patients with the mutation IVS3 + 10 T --> G. Among these patients, there were only three cases of patients with diabetes (9.37%). This was lower than the prevalence of abnormalities in the exons of the PRSS1 gene (51.92%): 12 patients with c.361 G --> A, eight patients with c.415 T --> A, and five patients with c.365G --> A. Among them were 12 persons with diabetes, including five requiring insulin to regulate blood sugar. What is more, among the 27 patients carrying PRSS1 gene polymorphism (c.486 C --> T, within the exon 4), there were 15 persons with diabetes symptoms. More than 40% of these patients required insulin to regulate blood sugar. CONCLUSIONS: An abnormality within the intron region of the PRSS1 gene represents one of the causes of pancreatitis in Chinese patients, but it is not related to pancreatic diabetes. However, the exon abnormality obviously raises the morbidity rate of type 3c diabetes, which relies on insulin. PMID- 20001682 TI - Effect of islet transplantation on metabolic glucose control in rats with diabetes. AB - BACKGROUND: Transplantation of pancreatic islets has been extensively investigated as a strategy for glycemic control in experimental animals and in patients with diabetes. We investigated whether islet transplantation allows us to obtain adequate islet function during glucose stimulation using a continuous glucose monitoring system (CGMS) in the rat. METHODS: We investigated four groups of eight rats each: healthy rats (controls), rats with diabetes, and rats with diabetes transplanted with microencapsulated islets in the peritoneal cavity or transplanted with free islets under the kidney capsule. Syngeneic islets were isolated from Lewis rats. After diabetes induction and islet implantation, when glycemia was stable, a glucose sensor was implanted, and an intraperitoneal glucose tolerance test (IPGTT) was performed to evaluate islet function. Interstitial glucose levels were analyzed, using a theoretical model, to estimate kinetics of glucose metabolism. RESULTS: Islet transplantation was effective in inducing normoglycemia in both groups, but results of IPGTTs showed that in animals with islets transplanted in microcapsules values of area under the curve and total glucose elimination constant (k(tot)) were significantly different from those in control animals and that these differences were even more important in animals with islets implanted under the kidney capsule. CONCLUSIONS: Our present investigation demonstrates that the application of CGMS was effective in evaluation of glucose metabolism by islet transplantation and indicates that efficient diabetes control can be achieved with this technology. PMID- 20001683 TI - A new model to estimate bolus insulin need. AB - BACKGROUND: Patients with type 1 diabetes usually use the carbohydrate (CHO) counting method to establish their bolus insulin need. However, most still struggle with blood glucose control. We believe that for good control their insulin requirements should strive to mimic the insulin secretion of those without diabetes. The objective here is to develop, from first principles, a better model than the CHO counting model for calculating the bolus insulin need of subjects without diabetes. Such a model may also provide better blood glucose control for patients with type 1 diabetes. This will be investigated in a future article. METHODS: Equations for metabolism of CHO and for insulin response were derived from first principles. Clinical trials were used to verify these models. The final results--namely, the new bolus insulin requirement equations--were verified using clinical trials by other researchers (Wolever and co-workers). Their methods used are described in their articles given in the list of references. RESULTS: The postprandial insulin secretion relationships resulted in an average Pearson R(2) of 0.807 (for the new method) versus the old method's R(2) of 0.562 (CHO counting). CONCLUSIONS: The newly derived equation provides a better approximation than the CHO counting method of insulin secretion due to metabolized blood glucose energy from ingested carbohydrates for those without diabetes. We believe that insulin dosage requirements for a patient with type 1 diabetes should mimic the insulin secretion of those without diabetes. If this is true, it means that the new equation should also estimate bolus insulin need for a patient with type 1 diabetes more accurately than before. This will be investigated in a future article. PMID- 20001684 TI - Accuracy of whole blood glucose measurement when venous catheter blood samples are used on glucose meters. AB - BACKGROUND: Previous studies have found positive bias and frequent outliers when central venous catheter (CVC) whole blood is used to dose glucose meters. We designed a study to determine whether positive bias and outliers with CVC whole blood glucose samples are due to exogenous glucose contamination of CVC samples, inherent bias and imprecision of glucose meters, or properties of CVC whole blood that interfere with the function of some glucose meters. METHODS: We studied the relationship between venous whole blood and venous plasma glucose drawn by venipuncture to CVC whole blood and CVC plasma glucose in 50 hospitalized patients. In 27 patients whole blood glucose was measured on both the Accu-Chek Inform (Roche Diagnostics, Indianapolis, IN) and StatStrip (Nova Biomedical, Waltham, MA). RESULTS: By comparing CVC plasma to venous plasma glucose, we determined that contamination of CVC samples with exogenous glucose was uncommon. On the Inform meter outliers were approximately twice as common with CVC whole blood compared to venous whole blood. In 27 patients who had CVC whole blood analyzed by both Inform and StatStrip, outliers occurred approximately twice as often on the Inform compared to the StatStrip. Accounting for CVC samples contaminated with exogenous glucose, outliers on the StatStrip did not occur significantly more often using CVC whole blood compared to venous whole blood. CONCLUSIONS: Properties unique to CVC whole blood differentially affect glucose meter bias and imprecision. Device selection is critical in practices that wish to use CVC whole blood to monitor glucose concentration in hospitalized patients. PMID- 20001686 TI - Diffusion tensor imaging at 3 hours after traumatic spinal cord injury predicts long-term locomotor recovery. AB - Accurate diagnosis of spinal cord injury (SCI) severity must be achieved before highly aggressive experimental therapies can be tested responsibly in the early phases after trauma. These studies demonstrate for the first time that axial diffusivity (lambda||), derived from diffusion tensor imaging (DTI) within 3 h after SCI, accurately predicts long-term locomotor behavioral recovery in mice. Female C57BL/6 mice underwent sham laminectomy or graded contusive spinal cord injuries at the T9 vertebral level (5 groups, n = 8 for each group). In-vivo DTI examinations were performed immediately after SCI. Longitudinal measurements of hindlimb locomotor recovery were obtained using the Basso mouse scale (BMS). Injured and spared regions of ventrolateral white matter (VLWM) were reliably separated in the hyperacute phase by threshold segmentation. Measurements of lambda|| were compared with histology in the hyperacute phase and 14 days after injury. The spared normal VLWM determined by hyperacute lambda|| and 14-day histology correlated well (r = 0.95). A strong correlation between hindlimb locomotor function recovery and lambda||-determined spared normal VLWM was also observed. The odds of significant locomotor recovery increased by 18% with each 1% increase in normal VLWM measured in the hyperacute phase (odds ratio = 1.18, p = 0.037). The capability of measuring subclinical changes in spinal cord physiology and murine genetic advantages offer an early window into the basic mechanisms of SCI that was not previously possible. Although significant obstacles must still be overcome to derive similar data in human patients, the path to clinical translation is foreseeable and achievable. PMID- 20001687 TI - Vascular endothelial growth factor is involved in mediating increased de novo hippocampal neurogenesis in response to traumatic brain injury. AB - Stimulating the endogenous repair process after traumatic brain injury (TBI) can be an important approach in neuroregenerative medicine. Vascular endothelial growth factor (VEGF) is one of the molecules that can increase de novo hippocampal neurogenesis. Here, we tested whether VEGF signaling through Flk1 (VEGF receptor 2) is involved in the neurogenic process after experimental TBI. We found that Flk1 is expressed both by neuroblasts in the subgranular layer (SGL) and by maturing granule neurons in the adult dentate gyrus (DG) of the hippocampus. After lateral fluid percussion TBI (LFP-TBI) in the rat, we detected elevated VEGF levels and also increased numbers of de novo neurons in the ipsilateral DG. To test the involvement of VEGF and Flk1 in the neurogenic process directly, we delivered recombinant VEGF or SU5416, an inhibitor to Flk1, into the ipsilateral cerebral ventricle of injured animals. We found that VEGF infusion significantly increased the number of BrdU+/Prox1+ new neurons, decreased the number of TUNEL+ cells, but did not change the number of BrdU+ newborn cells per se. Infusion with SU5416 caused no significant changes. Our results suggest that (a) VEGF is a part of the molecular signaling network that mediates de novo hippocampal neurogenesis after TBI; (b) VEGF predominantly mediates survival of de novo granule neurons rather than proliferation of neuroblasts in the injured brain; and (c) additional VEGF receptor(s) and/or other molecular mechanism(s) are also involved in mediating increased neurogenesis following injury. PMID- 20001689 TI - Methadone maintenance treatment for youth: experiences of clients, staff, and parents. AB - In order to better understand the components of effective treatment for the understudied population of opioid-dependent youth, separate focus groups were conducted in 2006 with clients, clinical staff, and clients' parents (total n = 22) at a novel methadone maintenance program for adolescents and young adults in suburban New York. Focus group sessions were audiotaped, transcribed, and analyzed for common themes. Clients, staff, and parents all reported that effective treatment for opioid addiction among young people is a long-term program, combines pharmacological and behavioral approaches, emphasizes a high degree of individual attention from staff, and incorporates clients' family members. The study's limitations were noted. PMID- 20001690 TI - Use of small incentives for increasing participation and reducing dropout in a family drug-use prevention program in a Spanish sample. AB - Poor participation rates represent one of the most serious problems facing family based drug-use prevention programs. Strategies involving incentives have been used to increase recruitment and retention of the target population of such interventions, but in Spain, such strategies for modifying behavior are unusual. The goal of the research was to study the use of small financial incentives (euro10 voucher) as a strategy to increase attendance and reduce dropout in a family drug-prevention program applied in the school context. Participants were 211 pupils (aged 12-13) and their parents. The results show that small financial incentives can be useful to increase the attendance of families in prevention programs and to reduce dropout. PMID- 20001688 TI - Effect of endogenous androgens on 17beta-estradiol-mediated protection after spinal cord injury in male rats. AB - Several groups have recently shown that 17beta-estradiol is protective in spinal cord injury (SCI). Testosterone can be aromatized to 17beta-estradiol and may increase estrogen-mediated protection. Alternatively, testosterone has been shown to increase excitotoxicity in models of central nervous system (CNS) injury. These experiments test the hypothesis that endogenous testosterone in male rats alters 17beta-estradiol-mediated protection by evaluating a delayed administration over a clinically relevant dose range and manipulating testicular derived testosterone. Adult male Sprague Dawley rats were either gonadectomized or left gonad-intact prior to SCI. SCI was produced by a midthoracic crush injury. At 30 min post SCI, animals received a subcutaneous pellet of 0.0, 0.05, 0.5, or 5.0 mg of 17beta-estradiol, released over 21 days. Hindlimb locomotion was analyzed weekly in the open field. Spinal cords were collected and analyzed for cell death, expression of Bcl-family proteins, and white-matter sparing. Post SCI administration of the 0.5- or 5.0-mg pellet improved hindlimb locomotion, reduced urinary bladder size, increased neuronal survival, reduced apoptosis, improved the Bax/Bcl-xL protein ratio, and increased white-matter sparing. In the absence of endogenous testicular-derived androgens, SCI induced greater apoptosis, yet 17beta-estradiol administration reduced apoptosis to the same extent in gonadectomized and gonad-intact male rats. These data suggest that delayed post-SCI administration of a clinically relevant dose of 17beta-estradiol is protective in male rats, and endogenous androgens do not alter estrogen mediated protection. These data suggest that 17beta-estradiol is an effective therapeutic intervention for reducing secondary damage after SCI in males, which could be readily translated to clinical trials. PMID- 20001691 TI - Validation of the alcohol use identification test in a prison sample living in the Arabian Gulf region. AB - The purpose of this study was to assess the utility of the Arabic translation of the Alcohol Use Disorders Identification Test (AUDIT) among a sample of male Muslim prisoners (N = 107) in Dubai, United Arab Emirates (UAE). Confirmatory factor analysis indicated a two-factor model to be the best fit of the data. Good internal reliability (alpha =.91) and predictive validity were also observed. While the limitations of the study are observed, these promising results suggest that the Arabic version of the AUDIT is a reliable and sound measure of alcohol use disorders among prisoners in the UAE. PMID- 20001692 TI - Patterns of ecstasy use and associated harm: results of a Brazilian online survey. AB - The aim of this study was to gather information about ecstasy users in Brazil, particularly on issues related to risks associated to the use of the drug, so as to offer a basis to prevention projects. A total of 1,140 Brazilian ecstasy users answered an online questionnaire from August 2004 to February 2005. Participants were predominantly young single heterosexual well-educated males from upper economical classes. A categorical regression with optimal scaling (CATREG) was performed to identify the risks associated with ecstasy use. "Pills taken in life" had a significant correlation with every investigated risk, particularly ecstasy dependence, unsafe sex, and polydrug use. "Gender," "sexual orientation," and "socioeconomic class" were not predictive of risk behavior. The Internet proved to be a useful tool for data collection. Given the recent increase in ecstasy availability in Brazil, a first prevention campaign directed toward the drug is urgent. At least in a preliminary Brazilian intervention, the campaign must be conducted at night leisure places, mainly frequented by youngsters from upper socioeconomic classes. The results do not call for information material with specific targets, such as gender or sexual orientation. The study's limitations have been noted. PMID- 20001693 TI - Using latent class and latent transition analysis to examine the transtheoretical model staging algorithm and sequential stage transition in adolescent smoking. AB - Data on stage of change was collected between 1997-1999 on 4125 adolescent nonsmokers and smokers from central England who completed two versions of the stage of change algorithm for smoking acquisition and cessation. The fit of the latent variable defined by the spiral of change to adolescents' responses to the staging questions was examined using latent class analysis. Whether a model of sequential transition across stages fitted the observed transitions was tested with latent transition analysis (LTA). There was reasonable support for the staging algorithm but no evidence of sequential stage transition. The research was funded by Cancer Research UK. PMID- 20001694 TI - Association between narcotic use and anabolic-androgenic steroid use among American adolescents. AB - Drawing on the data gathered in the 2006 Monitoring the Future study of American youth, the present research examines associations between use of narcotics and use of anabolic-androgenic steroids (AASs) among high-school seniors (n = 2,489). With independent measures and controls including sex, race, media exposure, socializing with friends, participation in recreational and school-sponsored sports, perceptions of drug use among professional athletes, and perceptions of steroid use among close friends, binary logistic regression analyses revealed significant associations between AAS use and the use of alcohol, crack cocaine, Vicodin, gamma-hydroxybutyrate (GHB), Ketamine, and Rohypnol. While use of both AASs and the narcotic drugs generally did not eclipse 5% of the sample, the numbers extend to many thousands in larger populations. Implications for health practitioners and recommendations for future research are offered. The study's limitations are noted. PMID- 20001695 TI - Workplace responsibility, stress, alcohol availability and norms as predictors of alcohol consumption-related problems among employed workers. AB - The objectives of this study were to determine the prevalence of alcohol use and problems among employed individuals in Alberta, Canada (N = 1,890), and to conduct a multivariate examination of predictors of alcohol consumption-related problems. General alcohol problems were identified by 10%, although very few workers described any specific work-related alcohol problems (1%). Structural equation modeling revealed that, as hypothesized, workplace alcohol availability predicted general alcohol problems. Job responsibility and workplace norms also predicted alcohol problems but only for men. Perceived work stress did not predict alcohol problems. Results support the development of interventions that focus on re-shaping alcohol use norms. PMID- 20001696 TI - Reducing adolescent use of harmful legal products: intermediate effects of a community prevention intervention. AB - PURPOSE: Preliminary results are presented from a feasibility study of a comprehensive community prevention intervention to reduce the use of inhalants and other harmful legal products (HLPs) among adolescents in three Alaskan frontier communities conducted in 2004-2007. The legal products used to get high include over-the-counter drugs, prescription drugs, and common household products. Community mobilization, environmental and school-based strategies were implemented to reduce access, enhance knowledge of risks, and improve assertiveness and refusal skills. METHODS: Pre- and post-intervention survey data were collected from 5-7th grade students from schools in three communities using standardized instruments to assess knowledge, assertiveness, refusal skills, perceived availability, and intent to use. The intervention consisted of community mobilization and environmental strategies to reduce access to HLPs in the home, at school, and through retail establishments. In addition, the ThinkSmart curriculum was implemented in classrooms among 5th grade students to increase the knowledge of harmful effects of HLPs and improve the refusal skills. Data were analyzed using hierarchical linear models that enable corrections for correlated measurement error. RESULTS: Significant increases in knowledge of harms related to HLP use and decreases in perceived availability of HLP products were observed. The environmental strategies were particularly effective in reducing the perceived availability of HLPs among 6th and 7th graders. DISCUSSION: Although limited by the absence of randomized control groups in this preliminary study design, the results of this study provide encouragement to pursue mixed strategies for the reduction of HLP use among young people in Alaskan frontier communities. PMID- 20001697 TI - Family structure and adolescent drug use: an exploration of single-parent families. AB - Data from the 2004 Monitoring the Future survey examined a nationally representative cross-sectional sample of 8th to 12th grade adolescents in rural and urban schools from across the United States (N = 37,507). Results found that drug use among daughters living with single fathers significantly exceeded that of daughters living with single mothers, while gender of parent was not associated with sons' usage. This distinction in adolescent drug use between mother-only versus father-only households is largely overlooked in contemporary studies. Factors responsible for variations in sons' and daughters' usage in single-parent families have important implications for future drug prevention efforts. PMID- 20001698 TI - Combining women's preferences and expert advice to design a tailored smoking cessation program. AB - We designed a patient-centered smoking cessation program for women in 2004/2005, incorporating women's preferences and expert opinion. Our four-step process included: (1) concept-development focus groups; (2) an expert panel; (3) concept testing focus groups, and (4) a pilot study. Data analyses occurred in 2004-2007. The new program offered options: the traditional Veterans Health Administration (VA) male-dominated program was the least selected option in the pilot study. Patients can be effectively involved in program development. The study's implications and limitations are noted. This research (conducted in Los Angeles, California) was funded by the American Legacy Foundation with additional VA support. PMID- 20001699 TI - Characterizing and comparing young adult intermittent and daily smokers. AB - We interviewed 732 smokers (from five US upper Midwestern states) via telephone in 2006 to examine young adult smoking patterns. We first defined two groups of intermittent smokers-low (who smoked for 1-14 days in the past 30 days) and high (who smoked for 15-29 days in the past 30 days), and then analyzed differences between these two groups and daily smokers. Low intermittent smokers were much less likely to consider themselves smokers, feel addicted, or smoke with friends than high intermittent smokers. Daily smokers were more likely to feel addicted and have trouble quitting smoking than high intermittent smokers. Implications, limitations, and ideas for future studies will be discussed. PMID- 20001700 TI - The comparison of methods for determination of humoral and cell-mediated immunity in patients with breast cancer. AB - We observed the immunological answer to antigens obtained from the human malignant breast tumor and from the blood of inbred mice strain C3H/H2K infected by LDH virus. We compared the modified ELISA method for humoral immunity with the leukocyte adherence inhibition (LAI) assay for cell-mediated immunity. The modified ELISA method is suitable for early diagnosing and monitoring antibodies in a malignant breast tumor simultaneously with senological examinations which include mammography and clinical examinations, because the antibodies are determined in a high number of samples by single application. PMID- 20001701 TI - Effects of Lac operon activation, deletion of the Yhha gene, and the removal of oxygen on the ultra-weak photon emission of Escherichia coli. AB - We observed a relation between gene activity and ultra-weak photon emission (UPE). By comparing the UPEs of E. coli with the LacI gene present and deleted we found that more gene activity produced higher UPE. This relation was further confirmed by studying the UPE of the E. coli with and without the Yhha gene. We interpreted that a higher aminoacyl t-RNA synthetase activity, which used ATP from the respiratory chain, could increase the emission. Satisfying the increased need of ATP by the E. coli through an increase of respiratory chain activity, which has reactive oxygen species (ROS) as a byproduct, results in a higher rate of photon emission. To ensure that oxygen is at the origin of this emission, we replaced the air by pure nitrogen. After 30 min, it was observed that the emission levels equaled the emission levels of the sterile medium. We could therefore conclude that the source of the photon emission would be affected by genetic activity and is oxygen related. PMID- 20001702 TI - The effects of mobile-phone electromagnetic fields on brain electrical activity: a critical analysis of the literature. AB - We analyzed the reports in which human brain electrical activity was compared between the presence and absence of radio-frequency and low-frequency electromagnetic fields (EMFs) from mobile phones, or between pre- and post exposure to the EMFs. Of 55 reports, 37 claimed and 18 denied an EMF-induced effect on either the baseline electro encephalogram (EEG), or on cognitive processing of visual or auditory stimuli as reflected in changes in event-related potentials. The positive reports did not adequately consider the family-wise error rate, the presence of spike artifacts in the EEG, or the confounding role of the two different EMFs. The negative reports contained neither positive controls nor power analyses. Almost all reports were based on the incorrect assumption that the brain was in equilibrium with its surroundings. Overall, the doubt regarding the existence of reproducible mobile-phone EMFs on brain activity created by the reports appeared to legitimate the knowledge claims of the mobile phone industry. However, it funded, partly or wholly, at least 87% of the reports. From an analysis of their cognitive framework, the common use of disclaimers, the absence of information concerning conflicts of interest, and the industry's donations to the principal EMF journal, we inferred that the doubt was manufactured by the industry. The crucial scientific question of the pathophysiology of mobile-phone EMFs as reflected in measurements of brain electrical activity remains unanswered, and essentially unaddressed. PMID- 20001703 TI - I. The little explored efficacy of magnetic fields in cancer treatment and postulation of the mechanism of action. AB - There are many reports of enhanced transcription and replication in different cell culture systems exposed to electromagnetic fields, and reports of cytoreduction (necrosis and apoptosis) in tumors transplanted into animals exposed to similar, often much stronger electromagnetic fields, but where heating is negligible. Although the mechanism of apoptosis has not been characterized, one major candidate for the initiation of such a process is the production of numerous breaks in DNA, and the overwhelming of DNA repair processes, leading to the initiation of the apoptotic process. In this article, we concentrate on the potential of strong magnetic fields to play a role in cancer treatment. PMID- 20001704 TI - II. Model building: an electrical theory of control of growth and development in animals, prompted by studies of exogenous magnetic field effects (paper I), and evidence of DNA current conduction, in vitro. AB - A theory of control of cellular proliferation and differentiation in the early development of metazoan systems, postulating a system of electrical controls "parallel" to the processes of molecular biochemistry, is presented. It is argued that the processes of molecular biochemistry alone cannot explain how a developing organism defies a stochastic universe. The demonstration of current flow (charge transfer) along the long axis of DNA through the base-pairs (the "pi way) in vitro raises the question of whether nature may employ such current flows for biological purposes. Such currents might be too small to be accessible to direct measurement in vivo but conduction has been measured in vitro, and the methods might well be extended to living systems. This has not been done because there is no reasonable model which could stimulate experimentation. We suggest several related, but detachable or independent, models for the biological utility of charge transfer, whose scope admittedly outruns current concepts of thinking about organization, growth, and development in eukaryotic, metazoan systems. The ideas are related to explanations proposed to explain the effects demonstrated on tumors and normal tissues described in Article I (this issue). Microscopic and mesoscopic potential fields and currents are well known at sub-cellular, cellular, and organ systems levels. Not only are such phenomena associated with internal cellular membranes in bioenergetics and information flow, but remarkable long-range fields over tissue interfaces and organs appear to play a role in embryonic development (Nuccitelli, 1992 ). The origin of the fields remains unclear and is the subject of active investigation. We are proposing that similar processes could play a vital role at a "sub-microscopic level," at the level of the chromosomes themselves, and could play a role in organizing and directing fundamental processes of growth and development, in parallel with the more discernible fields and currents described. PMID- 20001705 TI - Zero magnetic field effect observed in human cognitive processes. AB - In our previous works, we reported that compensation of the geomagnetic field to a level less than 0.4 microT ("zero magnetic field," or ZMF) affected human cognitive processes. ZMF exposure increased the number of errors and the task processing time by 2.4% in average. However, in the array of the magnetic effects calculated from the experimental data, some readings have been found to deviate from the mean magnetic effect by more than three standard deviations. This finding could give rise to doubt as to whether the magnetic effect observed was a mere sequence of the presence of such unlikely data values. In the present work we examine the results of the unlikely data elimination and show that the corrected magnetic effect in tested humans remains statistically significant, though at a reduced magnitude 1.5%. PMID- 20001706 TI - Frohlich's coherent excitations & the cancer problem--a retrospective overview of his guiding philosophy. AB - The guiding philosophy underlying Frohlich's approach to biology from the side of theoretical physics is summarized, and illustrated, in the context of his prediction of (dynamic) coherent excitations in living systems, based on their dielectric and elastic properties, and far-from-equilibrium (nonlinear) character. His envisaged role of these coherent excitations in cell division and its control is outlined, together with the associated implications for cancer--as understood both at the time of his work and subsequently. PMID- 20001707 TI - Cladribine inhibits a diltiazem-induced increase in red blood cell purine nucleotide concentrations in a zebrafish model. AB - Minimizing drug interactions is paramount to improving the efficacy and tolerability of cancer therapy. The zebrafish represents an innovative cancer model due to highly conserved genetics and inherent capacity for high-throughput chemical screening. This pilot study extends the utility of the zebrafish to a preclinical model for pharmacodynamics by examining the interaction of the nucleoside analogue, cladribine with the calcium channel blocker, diltiazem. Cladribine (0.7-3.5 mM) and/or diltiazem (2.4 mM), was injected intraperitoneally into adult zebrafish and red blood cell (RBC) lysates were assayed by HPLC for levels of purine nucleotides (e.g. ATP), potential biomarkers of cardiovascular health. Diltiazem increased RBC ATP concentrations, which were inhibited by co injection of cladribine. These results suggest a novel drug interaction and highlight the feasibility of the zebrafish as an in vivo model for pharmacodynamic studies. PMID- 20001708 TI - Comparison of three oxidative stress biomarkers in a sample of healthy adults. AB - Oxidative stress is a potentially important aetiological factor for many chronic diseases, including cardiovascular disease, neurodegenerative disease and cancer, yet studies often find inconsistent results. The associations between three of the most widely used biomarkers of oxidative stress, i.e. F(2)-isoprostanes for lipid peroxidation and 8-oxo-7,8-dihydro-2'-deoxyguanosine (8-oxo-dG) and the comet assay with FPG for oxidative DNA damage, were compared in a sample of 135 healthy African-American and white adults. Modest associations were observed between F(2)-isoprostanes and the comet assay (r = 0.22, p = 0.01), but there were no significant correlations between 8-oxo-dG and the comet assay (r = -0.09) or F(2)-IsoP (r = -0.04). These results are informative for researchers seeking to compare results pertaining to oxidative stress across studies and/or assessment methods in healthy disease-free populations. The development and use of oxidative stress biomarkers is a promising field; however, additional validation studies are necessary to establish accuracy and comparability across oxidative stress biomarkers. PMID- 20001709 TI - Cytokine array analysis of peritoneal fluid between women with endometriosis of different stages and those without endometriosis. AB - Cytokines are key mediators of intercellular communication and are likely to promote the development and progression of endometriosis. Previous studies provided evidence that endometriosis develops as a result of the pathogenetic factors in the peritoneal environment, especially the peritoneal fluid (PF). We determined different cytokine expression in peritoneal fluid between women with minimal/mild and moderate/severe endometriosis and those without endometriosis using the cytokine array. As a result, 78 cytokines were found to have a threefold change, including 74 increases and four decreases in endometriosis compared with the control group; 96 cytokines had a threefold change including 91 increases and five decreases in minimal and mild endometriosis compared with the control group; 83 cytokines had a threefold change including 14 increases and 69 decreases in moderate and severe endometriosis compared with minimal and mild endometriosis. The cytokine networks were produced by Pathway Studio software and revealed that most cytokines are involved in cell binding, interaction and protein synthesis and transportation regulation. Among them activin A, Smad7 and beta-nerve growth factor are the most interesting as they may be involved in the pathogenesis of endometriosis. These results suggest that cytokines are very important factors in the development of endometriosis. The findings of differentially expressed cytokines improves our knowledge of the pathogenesis and development of endometriosis and these findings warrant further studies to develop potential targets for the diagnosis and treatment of endometriosis. PMID- 20001710 TI - hTERT methylation and expression in gastric cancer. AB - Gastric cancer is the second most prevalent cause of cancer death worldwide. DNA methylation is a common event in gastric carcinogenesis. hTERT seems to be the rate-limiting determinant of telomerase activation, which is responsible for stability and life span. hTERT hypermethylation has been associated with telomerase expression. In the present study, we investigated the promoter methylation status and hTERT protein expression in gastric cancer and normal mucosa samples. One hundred and nine gastric cancer and 53 normal mucosa samples were investigated through methylation-specific PCR. Immunohistochemistry was analysed using peroxidase in 55 gastric cancer and 18 normal gastric mucosa samples. This is the first study evaluating hTERT methylation status in gastric carcinogenesis. We did not observe hTERT protein expression in normal gastric mucosa. Moreover, hTERT expression was observed in 80% of tumours and was associated with gastric cancer (p < 0.0001). Partial methylation was the most frequent pattern in gastric samples, even in normal mucosa. The frequency of specimens presenting hypermethylation was significantly higher in tumours than in normal mucosa samples (p = 0.0002), although the presence of hypermethylated promoter was not associated with a higher frequency of hTERT expression. A low correlation between hTERT protein expression and methylation was verified in gastric cancer samples. There was a clear difference in the frequency of hTERT expression and methylation within tumoral and non-tumoral tissues. Methylation status and telomerase expression may be useful for the diagnosis of gastric cancer and may have an impact on the anti-telomerase strategy for cancer therapy. PMID- 20001711 TI - Biomarkers in veterinary medicine: establishing a new international forum for veterinary biomarker research. PMID- 20001712 TI - Frontiers in thyroid cancer: December 2009. PMID- 20001713 TI - On the fortieth anniversary of thyrotropin-releasing hormone: the hormone that launched a new era. PMID- 20001714 TI - Thyroid stem cells and cancer. AB - BACKGROUND: Thyroid gland development and function are essential for life, and recent findings indicate the presence of stem/progenitor cells within the thyroid gland as a potential source of tissue regeneration and cancer formation. SUMMARY: This review summarizes the current knowledge on early differentiation of thyroid cells from embryonic stem cells and highlights exciting concepts and recent novel findings on adult thyroid stem/progenitor cells in the normal thyroid gland and in thyroid cancer. Other potential sources and markers of stem/progenitor cells in the thyroid include bone marrow, microchimerism, and embryological remnant derived multifocal solid cell nests. Finally, we discuss new therapeutic strategies that target thyroid cancer stem cells. CONCLUSIONS: Thyroid stem/progenitor cell populations are present in the normal and diseased thyroid gland. Advances in normal and cancer thyroid stem cell biology will be essential for future targeted therapies. PMID- 20001715 TI - Lessons from mouse models of thyroid cancer. AB - BACKGROUND: Thyroid cancer is the most common endocrine tumor and is increasing in incidence. The aim of this study was to review mouse models of differentiated thyroid cancer and how they elucidate human thyroid cancer biology. SUMMARY: Differentiated thyroid cancer, primarily papillary and follicular, comprises the majority of thyroid cancers. There has been tremendous growth in the cross-talk between basic science and clinical practice for thyroid cancer management. Insight into the framework of genes responsible for differentiated thyroid cancer has been gained through the use of mouse models. Common genetic alterations found in human papillary thyroid cancer such as RET/PTC rearrangements or the BRAF(V600E) mutation have genetically modified mouse counterparts. These and other preclinical mouse models have validated the importance of the cyclic adenosine monophosphate (cAMP)/protein kinase A and mitogen-activated protein kinase (MAPK) signaling pathways in papillary thyroid cancer (PTC). RAS mutations have a role in both papillary and follicular thyroid cancer development. Mice with overactivation of the phosphatidylinol-3-kinase (PI3K)-AKT and/or thyrotropin-regulated signaling pathways have been found to develop follicular thyroid cancer. Additional mouse models of thyroid cancer that utilize inducible expression systems are in development or are being characterized and will better reflect the majority of human thyroid cancers which are non-hereditary. Advances in in vivo imaging of mice allow for earlier detection of metastasis and the ability to follow tumor growth or regression which may be used in evaluation of pharmaceutical agents. CONCLUSIONS: Mouse models have expanded our understanding of the altered signaling pathways that contribute to thyroid cancer tumorigenesis and provide a powerful tool to develop novel diagnostic approaches and therapies. PMID- 20001717 TI - Clinical features and genetic predisposition to hereditary nonmedullary thyroid cancer. AB - BACKGROUND: Approximately 5% of the nonmedullary thyroid cancers are hereditary. Hereditary nonmedullary thyroid cancer may occur as a minor component of familial cancer syndromes (familial adenomatous polyposis, Gardner's syndrome, Cowden's disease, Carney's complex type 1, Werner's syndrome, and papillary renal neoplasia) or as a primary feature (familial nonmedullary thyroid cancer [FNMTC]). The goal of this article was to review our current knowledge on the hereditary nonmedullary thyroid cancer. SUMMARY: Epidemiologic and clinical kindred studies have demonstrated that FNMTC is a unique clinical entity. Most studies suggest that FNMTC is associated with more aggressive disease than sporadic cases, with higher rates of multicentric tumors, lymph node metastasis, extrathyroidal invasion, and shorter disease-free survival. A hereditary predisposition to nonmedullary thyroid cancer is well established, but the susceptibility genes for isolated FNMTC have not been identified. However, additional susceptibility loci for FNMTC have been recently identified in classic isolated cases of FNMTC (1q21, 6q22, 8p23.1-p22, and 8q24). CONCLUSIONS: More studies are needed to validate chromosomal susceptibility loci and identify the susceptibility genes for FNMTC. The discovery of the predisposing genes may allow for screening and early diagnosis, which could lead to improved outcomes for patients and their families. PMID- 20001718 TI - Advances in ultrasound for the diagnosis and management of thyroid cancer. AB - BACKGROUND: Ultrasonography (US) is increasingly being employed by endocrinologists and surgeons in the diagnosis and management of patients with thyroid nodules and thyroid cancer. Recent consensus guidelines from the American Thyroid Association highlight the importance of this modality by recommending that patients with thyroid nodules should undergo further evaluation with cervical US to stratify the risk of malignancy. Likewise, ultrasound is advocated for the preoperative and postoperative diagnosis of cervical lymph node metastases. SUMMARY: This article will summarize the US characteristics that impart a suspicious appearance on thyroid nodules and cervical lymph nodes as well as those findings that are reassuring. Likewise, the indications for thyroid nodule and lymph node fine-needle aspiration will be reviewed. Finally, this article will briefly discuss adjunctive tools in US such as elastography, percutaneous ethanol ablation, and radiofrequency ablation. CONCLUSIONS: US may be used to help stratify the risk of malignancy in thyroid nodules and cervical lymph nodes. This tool further aids in the diagnosis of malignancy when used in conjunction with fine-needle aspiration. US plays an important role as both a diagnostic and therapeutic tool in the evaluation of patients with neck masses. PMID- 20001716 TI - Potential utility and limitations of thyroid cancer cell lines as models for studying thyroid cancer. AB - BACKGROUND: Tumor-derived cell lines are widely used to study the mechanisms involved in thyroid carcinogenesis but recent studies have reported redundancy among thyroid cancer cell lines and identification of some "thyroid cell lines" that are likely not of thyroid origin. SUMMARY: In this review, we have summarized the uses, the limitations, and the existing problems associated with the available follicular cell-derived thyroid cancer cell lines. There are some limitations to the use of cell lines as a model to "mimic" in vivo tumors. Based on the gene expression profiles of thyroid cell lines originating from tumors of different types it has become apparent that some of the cell lines are closely related to each other and to those of undifferentiated carcinomas. Further, many cell lines have lost the expression of thyroid-specific genes and have altered karyotypes, while they exhibit activation of several oncogenes (BRAF, v-raf murine sarcoma viral oncogene homolog B1; RAS, rat sarcoma; and RET/PTC, rearranged in transformation/papillary thyroid carcinoma) and inactivation of tumor suppressor gene (TP53) which is known to be important for thyroid tumorigenesis. CONCLUSIONS: A careful selection of thyroid cancer cell lines that reflect the major characteristics of a particular type of thyroid cancer being investigated could be used as a good model system to analyze the signaling pathways that may be important in thyroid carcinogenesis. Further, the review of literature also suggests that some of the limitations can be overcome by using multiple cell lines derived from the same type of tumor. PMID- 20001719 TI - Controversies in the surgical management of newly diagnosed and recurrent/residual thyroid cancer. AB - BACKGROUND: Thyroid surgery is the mainstay of treatment for thyroid cancer, but there are options for the surgical approach. There is a general agreement that the goal of thyroid cancer surgery is to remove evident disease, minimize treatment and disease-related morbidity, minimize the risk of recurrence and metastatic spread, and facilitate long-term follow-up. Within these parameters, however, there are a number of controversies regarding the extent of surgery. SUMMARY: To minimize the risk of recurrence and to optimize the treatment of patients with thyroid cancer, a total thyroidectomy should be performed as the initial operation if the preoperative diagnosis is consistent with thyroid cancer. Macroscopic lymph node involvement does impact recurrence and may influence survival. Therefore, a preoperative ultrasound evaluation of the neck is essential before operative intervention. If pathologic nodes are identified in either the central or lateral neck they should be removed at the initial operation. The role of prophylactic central neck dissection remains controversial, and the risks of this procedure may outweigh the benefits. Therefore, we do not recommend the routine performance of a central neck dissection. Similar to the recent American Thyroid Association guidelines, we agree that their may be a subset of high-risk patients who may benefit from a prophylactic central neck dissection, but that population has yet to be defined. The extent of lateral dissection that is required is also debated as levels 1 and 5 lymph node involvement is uncommon with thyroid cancer. Recurrent disease can be challenging to remove, but several adjuncts are available to maximize the safety and efficacy of reoperation, making it the best option for patients with localized recurrences. CONCLUSIONS: Total thyroidectomy and neck dissection are very safe procedures when performed by experienced surgeons and can play an essential role in the management of patients with thyroid cancer. PMID- 20001720 TI - The benefits and risks of I-131 therapy in patients with well-differentiated thyroid cancer. AB - BACKGROUND: I-131 has been used in the therapy of well-differentiated thyroid cancer for over 50 years. Although the benefits and risks of I-131 remain issues of controversy and research, our understanding of them continues to improve. This review presents an overview of the benefits of I-131 therapy for ablation, adjuvant treatment, and treatment of locoregional and/or metastasis of well differentiated thyroid cancer and considers the risks of complications of I-131 therapy. SUMMARY: The benefits of I-131 remnant ablation include: [1] facilitating the interpretation of subsequent serum thyroglobulin levels, [2] increasing the sensitivity of detection of locoregional and/or metastatic disease on subsequent follow-up radioactive iodine whole-body scans, [3] maximizing the therapeutic effect of subsequent treatments, and [4] allowing a postablation scan to help identify additional sites of disease that were not identified on the preablation scan or when a preablation scan was not performed. The potential benefits of I-131 adjuvant treatment include decreasing recurrence and disease specific mortality for unknown microscopic, locoregional, and/or distant metastatic disease. The potential benefits of I-131 treatment of known locoregional and/or distant metastases are [1] decreasing recurrence, and [2] decreasing disease-specific mortality and/or palliation. The more significant risks and side effects involve organ systems including eye/nasolacrimal, salivary, pulmonary, gastrointestinal, hematopoietic, and gonads as well as secondary primary malignancies. CONCLUSIONS: Although there are never-ending controversies regarding I-131 therapy in well-differentiated thyroid cancer, the benefits and risks are becoming better understood. This in turn helps the treating physician and patient in making decisions regarding therapy. PMID- 20001721 TI - Clinical trials for progressive differentiated thyroid cancer: patient selection, study design, and recent advances. AB - Treatment of metastatic differentiated thyroid cancer is first based on the use of radioiodine and thyrotropin-suppressive thyroid hormone treatment. The recent availability of molecular-targeted therapies has lead to reconsideration of the treatment strategy in differentiated thyroid cancer patients with distant metastases who are resistant to radioiodine therapy. In those with progressive disease, treatment with kinase inhibitors should be offered preferably in the context of a prospective trial. PMID- 20001722 TI - Perfluorooctane sulfonate and perfluorooctanoic acid in surgical thyroid specimens of patients with thyroid diseases. AB - BACKGROUND: Perfluorooctanoic acid (PFOA) and perfluorooctane sulfonate (PFOS) are ubiquitous compounds that may act as endocrine disruptors, neurotoxic agents, and fetal development perturbing substances and may also be carcinogenic, as recently demonstrated in experimental animal models. There is little information on the potential for these compounds to affect the thyroid. Therefore, this study was performed to measure the intrathyroidal levels of PFOA and PFOS in surgical specimens of thyroid glands and to determine if there was a relationship between the concentrations of these substances and the clinical, biochemical, and histologic phenotype of the patients from whom the thyroids were obtained. We also sought to determine if there was a relationship between tissue and serum levels of both PFOA and PFOS. METHODS: PFOA and PFOS were measured in 28 patients undergoing thyroid surgery for benign (15 multinodular goiters and 7 Graves' disease) and malignant (5 papillary and 1 follicular carcinoma) thyroid disorders. RESULTS: PFOA and PFOS were detectable in all surgical specimens of thyroid tissue. Their median concentrations were 2.0 ng/g (range = 0.4-4.6 ng/g) and 5.3 ng/g (range = 2.1-44.7), respectively. Intrathyroidal concentrations of PFOA and PFOS were similar in the thyroids of patients with thyroid diseases as in thyroid glands obtained at autopsy. There was no relationship between the intrathyroidal concentrations of either PFOA or PFOS and the underlying thyroid disease. A significant correlation between the serum and the tissue levels of PFOS was found in all patients. The serum concentrations of PFOA and PFOS were significantly higher than those in the correspondent surgical specimens. CONCLUSIONS: These observations do not support the view that PFOA and PFOS are actively concentrated in the thyroid. PFOA and PFOS, however, are both found in surgical and autopsy thyroid specimens. Therefore, further studies to determine if they have disrupting effects in thyroid cells or tissue, and studies to compare populations with and without these compounds in their thyroid glands, are important. PMID- 20001723 TI - Thyrotoxic periodic paralysis presenting as polymorphic ventricular tachycardia induced by painless thyroiditis. PMID- 20001727 TI - Treatment of panic. AB - The present review summarizes empirical developments in the psychosocial treatment of panic and panic disorder by focusing on four key themes that have been emphasized in this literature: (a) treatment efficacy, effectiveness, and innovation; (b) modulating and process variables in treatment; (c) effective dissemination of treatment; and (d) prevention of panic. Cognitive behavioral treatments (CBTs) for panic are very good, even in the context of comorbidity, but the level of efficacy does not appear to be increasing over time. Unfortunately, there have been relatively few substantive innovations in CBT protocols since the 1980s, and despite progress in the use of computer technology, dissemination of these treatments is lagging. The prevention of panic spectrum psychopathology is a promising area linked with advances in the identification of panic-relevant risk factors that appear to modulate CBT outcomes. PMID- 20001726 TI - Timing of decompressive surgery of spinal cord after traumatic spinal cord injury: an evidence-based examination of pre-clinical and clinical studies. AB - While the recommendations for spine surgery in specific cases of acute traumatic spinal cord injury (SCI) are well recognized, there is considerable uncertainty regarding the role of the timing of surgical decompression of the spinal cord in the management of patients with SCI. Given this, we sought to critically review the literature regarding the pre-clinical and clinical evidence on the potential impact of timing of surgical decompression of the spinal cord on outcomes after traumatic SCI. The primary literature search was performed using MEDLINE, CINAHL, EMBASE, and Cochrane databases. A secondary search strategy incorporated articles referenced in prior meta-analyses and systematic and nonsystematic review articles. Two reviewers independently assessed every study with regard to eligibility, level of evidence, and study quality. Of 198 abstracts of pre clinical studies, 19 experimental studies using animal SCI models fulfilled our inclusion and exclusion criteria. Despite some discrepancies in the results of those pre-clinical studies, there is evidence for a biological rationale to support early decompression of the spinal cord. Of 153 abstracts of clinical studies, 22 fulfilled the inclusion and exclusion criteria. While the vast majority of the clinical studies were level-4 evidence, there were two studies of level-2b evidence. The quality assessment scores varied from 7 to 25 with a mean value of 12.41. While 2 of 22 clinical studies assessed feasibility and safety, 20 clinical studies examined efficacy of early surgical intervention to stabilize and align the spine and to decompress the spinal cord; the most common definitions of early operation used 24 and 72 h after SCI as timelines. A number of studies indicated that patients who undergo early surgical decompression can have similar outcomes to patients who received a delayed decompressive operation. However, there is evidence to suggest that early surgical intervention is safe and feasible and that it can improve clinical and neurological outcomes and reduce health care costs. Based on the current clinical evidence using a Delphi process, an expert panel recommended that early surgical intervention should be considered in all patients from 8 to 24 h following acute traumatic SCI. PMID- 20001728 TI - Pathological narcissism and narcissistic personality disorder. AB - We review the literature on pathological narcissism and narcissistic personality disorder (NPD) and describe a significant criterion problem related to four inconsistencies in phenotypic descriptions and taxonomic models across clinical theory, research, and practice; psychiatric diagnosis; and social/personality psychology. This impedes scientific synthesis, weakens narcissism's nomological net, and contributes to a discrepancy between low prevalence rates of NPD and higher rates of practitioner-diagnosed pathological narcissism, along with an enormous clinical literature on narcissistic disturbances. Criterion issues must be resolved, including clarification of the nature of normal and pathological narcissism, incorporation of the two broad phenotypic themes of narcissistic grandiosity and narcissistic vulnerability into revised diagnostic criteria and assessment instruments, elimination of references to overt and covert narcissism that reify these modes of expression as distinct narcissistic types, and determination of the appropriate structure for pathological narcissism. Implications for the fifth edition of the Diagnostic and Statistical Manual of Mental Disorders and the science of personality disorders are presented. PMID- 20001729 TI - The genetics of mood disorders. AB - Studying the genetics of mood disorders has never been more exciting. We have moved rapidly from establishing the genetic basis of depression to asking questions about how genes are expressed. This has been made possible by the capacity to collect and sequence DNA for large samples cheaply. But "multidisciplinary" approaches investigating interrelationships between risk factors have also been increasingly adopted, encouraging collaborations between those studying genes and those studying the brain, cognition, and/or the social environment. In this review, we first describe findings from quantitative and molecular studies investigating the genetic basis of depression. Second, we present overviews of three hot topics of genetic research: gene-environment interplay, which considers how genetic factors shape exposure and responses toward the social environment; endophenotypic research, which identifies neurophysiological and psychological mediators of genetic risk; and epigenetics, which explain how early environments can foster changes in gene expression, altering subsequent emotional development. PMID- 20001730 TI - Polymorphisms in the vitamin A receptor and innate immunity genes influence the antibody response to rubella vaccination. AB - BACKGROUND: Genetic polymorphisms play an important role in rubella vaccine induced immunity. METHODS: We genotyped 714 healthy children after 2 age appropriate doses of rubella-containing vaccine for 142 potential single nucleotide polymorphisms (SNPs). RESULTS: Specific polymorphisms in the vitamin A receptor, retinoic acid-inducible gene I (RIG-I), and tripartite motif 5 and 22 (TRIM5 and TRIM22) genes were significantly associated with rubella vaccine humoral immunity. The minor allele of the rs4416353 in the vitamin A receptor gene was associated with an allele dose-related decrease (P = .019) in rubella antibody response. The minor allele of rs6793694, in the vitamin A receptor gene, was associated with an allele dose-related antibody decrease (p = .039). The minor variant of nonsynonymous SNP rs10813831 (Arg7Cys) in the RIG-I gene was associated with an allele dose-related decrease in rubella antibody level from 37.4 to 28.0 IU/mL (P = .035), whereas increased representation of the minor allele of the 5'UTR SNP (rs3824949, P = .015) in the antiretroviral TRIM5 gene was associated with an allele dose-related increase in rubella antibody. It is of particular interest that the nonsynonymous SNP rs3740996 (His43Tyr) in the TRIM5 gene was associated with variations in rubella antibody response (P = .016) after having been previously found to play a significant functional role. CONCLUSIONS: These findings further expand our immunogenetic understanding of mechanisms of rubella vaccine-induced immunity. PMID- 20001731 TI - Description of an influenza vaccination campaign and use of a randomized survey to determine participation rates. AB - OBJECTIVES: To describe the procedures used during an influenza immunization program and the use of a randomized survey to quantify the vaccination rate among healthcare workers with and without patient contact. DESIGN: Influenza immunization vaccination program and a randomized survey. SETTING: Johns Hopkins University and Health System. METHODS: The 2008/2009 Johns Hopkins Influenza Immunization Program was administered to 40,000 employees, including 10,763 healthcare workers. A 10% randomized sample (1,084) of individuals were interviewed to evaluate the vaccination rate among healthcare workers with direct patient contact. RESULTS: Between September 23, 2008, and April 30, 2009, a total of 16,079 vaccinations were administered. Ninety-four percent (94.5%) of persons who were vaccinated received the vaccine in the first 7 weeks of the campaign. The randomized survey demonstrated an overall vaccination rate of 71.3% (95% confidence interval, 68.6%-74.0%) and a vaccination rate for employees with direct patient contact of 82.8% (95% confidence interval, 80.1%-85.5%). The main reason (25.3%) for declining the program vaccine was because the employee had received documented vaccination elsewhere. CONCLUSIONS: The methods used to increase participation in the recent immunization program were successful, and a randomized survey to assess participation was found to be an efficient means of evaluating the workforce's level of potential immunity to the influenza virus. PMID- 20001732 TI - Clinical and economic impact of methicillin-resistant Staphylococcus aureus colonization or infection on neonates in intensive care units. AB - OBJECTIVE: The rising incidence and mortality of methicillin-resistant Staphylococcus aureus (MRSA) colonization or infection in children has become a great concern. This study aimed to determine the clinical and economic impact of MRSA colonization or infection on infants and to measure excess mortality, length of stay, and hospital charges attributable to MRSA. DESIGN: This is a retrospective cohort study. SETTING AND PATIENTS: The study included infants admitted to a level III-IV neonatal intensive care unit from September 1, 2004, through March 31, 2008. METHODS: A time-dependent proportional hazard model was used to analyze the association between MRSA colonization or infection and mortality. The relationships between MRSA colonization or infection and length of stay and between MRSA colonization or infection and hospital charges were assessed using a matched cohort study design. RESULTS: Of 2,280 infants, 191 (8.4%) had MRSA colonization or infection. Of 132 MRSA isolates with antibiotic susceptibility results, 106 were resistant to clindamycin and/or trimethoprim sulfamethoxazole, thus representing a noncommunity phenotype. The mortality rate was 17.8% for patients with MRSA colonization or infection and 11.5% for control subjects. Neither MRSA colonization (hazard ratio [HR], 0.9 [95% confidence interval {CI}, 0.5-1.5]; P > .05 ) nor infection (HR, 1.2 [95% CI, 0.7-1.9]; P > .05 ) was associated with increased mortality risk. Infection caused by MRSA strains that were resistant to clindamycin and/or trimethoprim-sulfamethoxazole increased the mortality risk by 40% (HR, 1.4 [95% CI, 0.9-2.2]; P > .05 ), compared with the mortality risk of control subjects, but the increase was not statistically significant. MRSA infection independently increased length of stay by 40 days (95% CI, 34.2-45.6; P < .001) and was associated with an extra charge of $164,301 (95% CI, $158,712-$169,889; P < .001). CONCLUSIONS: MRSA colonization or infection in infants is associated with significant morbidity and financial burden but is not independently associated with increased mortality. PMID- 20001733 TI - A cluster of bloodstream infections caused by Streptococcus gallolyticus subspecies pasteurianus that involved 5 preterm neonates in a university hospital during a 2-month period. PMID- 20001734 TI - Redox control of endoplasmic reticulum function. AB - The lumen of the endoplasmic reticulum constitutes a separate intracellular compartment with a special proteome and metabolome. The redox conditions of the organelle are also characteristically different from those of the other subcellular compartments. The luminal environment has been considered more oxidizing than the cytosol due to the presence of oxidative protein folding. However, recent observations suggest that redox systems in reduced and oxidized states are present simultaneously. The concerted action of membrane transporters and oxidoreductase enzymes maintains the oxidized state of the thiol-disulfide and the reduced state of the pyridine nucleotide redox systems, which are prerequisites for the normal redox reactions localized in the organelle. The powerful thiol-oxidizing machinery of oxidative protein folding continuously challenges the local antioxidant defense. Alterations of the luminal redox conditions, either in oxidizing or reducing direction, affect protein processing, are sensed by the accumulation of misfolded/unfolded proteins, and may induce endoplasmic reticulum stress and unfolded protein response. The activated signaling pathways attempt to restore the balance between protein loading and processing and induce programmed cell death if these attempts fail. Recent findings strongly support the involvement of redox-based endoplasmic reticulum stress in a plethora of human diseases, either as causative agents or as complications. PMID- 20001735 TI - Sialoblastoma: utility of Ki-67 and p53 as a prognostic tool and review of literature. AB - Sialoblastoma is a rare tumor of the salivary gland that commonly occurs in the parotid gland and occasionally in the sub-mandibular gland. The malignant potential of sialoblastoma has been documented in only 3 of 32 cases of sialoblastoma reported thus far. In the last 15 years, we have encountered 2 cases of sialoblastoma, in a newborn and in a 15-year-old boy, both arising within the parotid gland. Case 1 has been previously reported and although there were 2 recurrences, at 1 and 9 years post resection, it has shown benign biological behavior. Case 2 is unusual since the patient presented with metastases. We reviewed the 2 cases, including the 2 recurrences from the first case, for histologic and immunohistochemical differences. Although both cases showed similar cytomorphologic features, there was a significant difference in Ki 67 expression: 20% in case 1 (original tumor), <2% in case 1 (recurrent tumor), and nearly 70-80% in the recent malignant case. The difference is remarkable when combined with p53 expression, which was focally positive in the first case but diffusely positive in the second. This report highlights the potential utility of proliferation markers such as Ki-67 in concert with p53 expression to better predict the biological behavior of a rare but locally aggressive neoplasm. PMID- 20001736 TI - Changes in Neisseria meningitidis disease epidemiology in the United States, 1998 2007: implications for prevention of meningococcal disease. AB - BACKGROUND: In January 2005, a quadrivalent (serogroups A, C , Y, and W-135) meningococcal conjugate vaccine was licensed for use in adolescents. This report describes the epidemiologic features of meningococcal disease in the United States from January 1998 through December 2007, before and during implementation of adolescent quadrivalent meningococcal conjugate vaccination. METHODS: Data were collected from active surveillance for invasive Neisseria meningitidis conducted through the Active Bacterial Core surveillance (ABCs) sites during 1998 2007. Isolates from cases were serogrouped at the ABCs site and confirmed at the Centers for Disease Control and Prevention. Estimates of the incidence and number of cases in the 50 states were calculated, standardizing for race and age group. RESULTS: In the period 1998-2007, a total of 2262 cases of meningococcal disease were reported from ABCs sites; 11.3% of these cases were fatal. The estimated United States average annual incidence of meningococcal disease was 0.53 cases per 100,000 population (95% confidence interval, 0.51-0.55), and an estimated 1525 (95% confidence interval, 1470-1598) cases occurred annually. The annual incidence decreased 64.1%, from 0.92 cases per 100,000 population in 1998 to 0.33 cases per 100,000 population in 2007. Infants aged <1 year have the highest incidence of meningococcal disease (5.38 cases per 100,000 population). After introduction of the quadrivalent meningococcal conjugate vaccine, no significant decrease in serogroup C or Y meningococcal disease was seen among those aged 11 19 years in 2006-2007, compared with 2004-2005. CONCLUSIONS: Before the introduction of the quadrivalent meningococcal conjugate vaccine, the incidence of meningococcal disease in the United States decreased to a historic low. However, meningococcal disease still causes a substantial burden of disease among all age groups. Future vaccination strategies may include targeting infants and preventing serogroup B meningococcal disease. PMID- 20001737 TI - Conjugate meningococcal vaccines: lifesaving or inefficient use or both-no easy answer. PMID- 20001739 TI - Laparoscopic surgery in 46,XX disorder of sex development: hysterosalpingectomy with gonadectomy. AB - PURPOSE: We present the outcomes of one of the largest series specifically of laparoscopic hysterosalpingectomy with bilateral gonadectomy in 46,XX patients with congenital adrenal hyperplasia raised as a male. PATIENTS AND METHODS: From June 2005 to March 2008, five patients raised as male were treated at our institution using laparoscopic surgery. 46,XX disorder of sex development was diagnosed in all the patients because of congenital adrenal hyperplasia. Hysterosalpingectomy with bilateral gonadectomy was performed completely laparoscopically in all five patients. RESULTS: All procedures were completed with minimal blood loss. The duration of the surgeries was 70-125 minutes. There were no complications during surgery or conversion to open surgery. The hospital stay ranged from 1 to 2 days, except in one patient who presented urinary retention and was discharged from the hospital a week after the surgery. CONCLUSIONS: Laparoscopic surgery can be safely used as part of the diagnosis and treatment of 46,XX disorder of sex development. Laparoscopy can be useful in the diagnosis as well as surgical management of Mullerian structures as well as intraabdominal gonads contrary to social sex. PMID- 20001740 TI - Designed three-dimensional collagen scaffolds for skin tissue regeneration. AB - One of the challenges in tissue engineering is the development of a reproducible three-dimensional (3D) scaffold to support cell migration and infiltration. As a dermal substitute, 3D collagen scaffolds with precisely controlled pore structures were fabricated using an innovative cryogenic dispenser system. The scaffolds were composed of perpendicular, highly porous collagen strands in successive layers. The fabricated scaffolds were evaluated in an in vitro keratinocyte/fibroblast coculture test. Fibroblasts were well dispersed within the scaffold, and keratinocytes had completely migrated through the well-designed pore structure and differentiated on top of the scaffold surface. The differentiated keratinocytes generated a stratum corneum in the 3D dispensed scaffolds, similar to that in normal skin tissue. PMID- 20001738 TI - Three-dimensional culture alters primary cardiac cell phenotype. AB - The directed formation of complex three-dimensional (3D) tissue architecture is a fundamental goal in tissue engineering and regenerative medicine. The growth of cells in 3D structures is expected to influence cellular phenotype and function, especially relative cell distribution, expression profiles, and responsiveness to exogenous signals; however, relatively few studies have been carried out to examine the effects of 3D reaggregation on cells from critical target organs, like the heart. Accordingly, we cultured primary cardiac ventricular cells in a 3D model system using a serum-free medium to test the hypothesis that expression profiles, multicellular organizational pathways, tissue maturation markers, and responsiveness to hormone stimulation were significantly altered in stable cell populations grown in 3D versus 2D culture. We found that distinct multi-cellular structures formed in 3D in conjunction with changes in mRNA expression profile, up-regulation of endothelial cell migratory pathways, decreases in the expression of fetal genes (Nppa and Ankrd1), and increased sensitivity to tri-iodothyronine stimulation when compared to parallel 2D cultures comprising the same cell populations. These results indicate that the culture of primary cardiac cells in 3D aggregates leads to physiologically relevant alterations in component cell phenotype consistent with cardiac ventricular tissue formation and maturation. PMID- 20001741 TI - Prevalence, molecular characterization, and phenotypic confirmation of extended spectrum beta-lactamases in Escherichia coli, Klebsiella pneumoniae, and Klebsiella oxytoca at the Radboud University Nijmegen Medical Centre in The Netherlands. AB - The prevalence and molecular types of extended-spectrum beta-lactamases (ESBLs) were determined during a 1-year period in unselected clinical nonduplicate isolates of Escherichia coli (n = 1,738), Klebsiella pneumoniae (n = 436), and Klebsiella oxytoca (n = 208), cultured at the University Medical Centre Nijmegen, The Netherlands. Isolates identified as ESBL producer by the Phoenix automated system were collected prospectively and subjected to molecular analysis for the most common ESBLs TEM, SHV, and CTX-M, as well as OXA and GES. Both the Etest ESBL and double-disk synergy test were performed as confirmatory tests. The estimated prevalence of ESBLs was 2.1% in E. coli, 5.2% in K. pneumoniae, and 2.4% in K. oxytoca. TEM-12 and -26, SHV-5 and -12, and CTX-M groups 1 and 9 were the most frequent ESBLs found. Isolates identified as ESBLs by the Phoenix were confirmed by polymerase chain reaction (PCR) in only 42%. In ESBL PCR-positive E. coli and K. pneumoniae, both confirmatory tests were positive in 95% of the isolates. In 28% of the Etest and 13% of the double-disk synergy test-positive isolates, PCR could not detect any ESBL gene. In these cases, other resistance mechanisms may play a role. Confirmatory tests were unreliable for K. oxytoca. A previously described mutation in the K1 enzyme was detected in one ceftazidime resistant K. oxytoca. The prevalence of ESBLs in The Netherlands is increasing. The predominant molecular types of ESBLs detected were comparable to other studies. Phoenix ESBL results need to be confirmed as advocated by ESBL detection guidelines. PMID- 20001742 TI - Microarray analysis of efflux pump genes in multidrug-resistant Mycobacterium tuberculosis during stress induced by common anti-tuberculous drugs. AB - Treatment of multidrug-resistant tuberculosis has become one of the major problems in public health. Understanding the molecular mechanisms of drug resistance has been central to tuberculosis research in recent times. DNA microarray technology provides the platform to study the genomic variations related to these mechanisms on a comprehensive level. To investigate the role of efflux pumps in drug resistance, we have constructed a custom DNA microarray containing 25 drug efflux pump genes of Mycobacterium tuberculosis (Indian Patent file no. 2071/DEL/2007) and monitored changes in the expression of these genes on exposure of common anti-tuberculous drugs. Expression profiling of efflux pump genes in multidrug-resistant M. tuberculosis isolates showed overexpression of 10 genes following exposure to various anti-tuberculous drugs. Although two of these genes (Rv3065 and Rv2938) have already been reported to be active drug efflux pumps in M. tuberculosis in earlier studies, the increased activities of other eight efflux pump genes (Rv1819, Rv2209, Rv2459, Rv2477c, Rv2688, Rv2846, Rv2994, and Rv3728) have been demonstrated in multidrug-resistant isolates by us for the first time. After confirmation of differential expressions of these genes by real time reverse transcription polymerase chain reaction, it was observed that a simultaneous overexpression of efflux pump genes Rv2459, Rv3728, and Rv3065 was associated with resistance to the combination of isoniazid and ethambutol, and these drugs, along with streptomycin, were identified to group together, where efflux-mediated drug resistance appears to be important in M. tuberculosis and follows a constant pattern of induction in multidrug-resistant isolates. Isoniazid and ethambutol combination was also found to be affected in 10% (6/60) of the clinical isolates in the presence of carbonyl cyanide m-chloro phenylhydrazone in resazurin microtitre plate assay, supporting the role of efflux pumps in the resistance to these drugs. Overexpression of two of the genes (Rv2477 and Rv2209) has also been observed with ofloxacin stress in M. tuberculosis. PMID- 20001743 TI - Using isoprostanes as biomarkers of oxidative stress: some rarely considered issues. AB - The measurement of F2-isoprostanes by methods utilizing mass spectrometry is widely regarded as the best currently available biomarker of lipid peroxidation. F2-isoprostanes and their metabolites can be measured accurately in plasma, urine, and other body fluids using mass spectrometric techniques, and detailed protocols have been published in several papers. However, many clinical studies and intervention studies with diets or supplements, have employed single "spot" measurements of F2-isoprostanes on either plasma/serum or urine to estimate "oxidative stress." This review examines the validity of the common assumption that plasma and urinary F2-isoprostane measurements are equivalent. It identifies scenarios where they may not be and where "spot" measurements can be misleading, with examples from the literature. We also discuss the controversial issue of whether and how F2-isoprostane levels in plasma should be standardized against lipids, and, if so, which lipids to use. PMID- 20001745 TI - Reciprocal regulation of Rac1 and PAK-1 by HIF-1alpha: a positive-feedback loop promoting pulmonary vascular remodeling. AB - Pulmonary vascular remodeling associated with pulmonary hypertension is characterized by media thickening, disordered proliferation, and in situ thrombosis. The p21-activated kinase-1 (PAK-1) can control growth, migration, and prothrombotic activity, and the hypoxia-inducible transcription factor HIF-1alpha was associated with pulmonary vascular remodeling. Here we studied whether PAK-1 and HIF-1alpha are linked in pulmonary vascular remodeling. PAK-1 was expressed in the media of remodeled pulmonary vessels from patients with pulmonary vasculopathy and was upregulated, together with its upstream regulator Rac1 and HIF-1alpha in lung tissue from lambs with pulmonary vascular remodeling. PAK-1 and Rac1 were activated by thrombin involving calcium, thus resulting in enhanced generation of reactive oxygen species (ROS) in human pulmonary artery smooth muscle cells (PASMCs). Activation of PAK-1 stimulated HIF activity and HIF-1alpha expression involving ROS and NF-kappaB, enhanced the expression of the HIF-1 target gene plasminogen activator inhibitor-1, and stimulated PASMC proliferation. Importantly, HIF-1 itself bound to the Rac1 promoter and enhanced Rac1 and PAK-1 transcription. Thus, PAK-1 and its activator Rac1 are novel HIF-1 targets that may constitute a positive-feedback loop for induction of HIF-1alpha by thrombin and ROS, thus explaining elevated levels of PAK-1, Rac1, and HIF 1alpha in remodeled pulmonary vessels. PMID- 20001746 TI - Receptor activation of NADPH oxidases. AB - Reactive oxygen species (ROS) have been implicated in many intra- and intercellular processes. High levels of ROS are generated as part of the innate immunity in the respiratory burst of phagocytic cells. Low levels of ROS, however, are generated in a highly controlled manner by various cell types to act as second messengers in redox-sensitive pathways. A NADPH oxidase has been initially described as the respiratory burst enzyme in neutrophils. Stimulation of this complex enzyme system requires specific signaling cascades linking it to membrane-receptor activation. Subsequently, a family of NADPH oxidases has been identified in various nonphagocytic cells. They mainly differ in containing one out of seven homologous catalytic core proteins termed NOX1 to NOX5 and DUOX1 or 2. NADPH oxidase activity is controlled by regulatory subunits, including the NOX regulators p47phox and p67phox, their homologs NOXO1 and NOXA1, or the DUOX1 or 2 regulators DUOXA1 and 2. In addition, the GTPase Rac modulates activity of several of these enzymes. Recently, additional proteins have been identified that seem to have a regulatory function on NADPH oxidase activity under certain conditions. We will thus summarize molecular pathways linking activation of different membrane-bound receptors with increased ROS production of NADPH oxidases. PMID- 20001747 TI - ER targeting and retention of the HCV NS4B protein relies on the concerted action of multiple structural features including its transmembrane domains. AB - The Hepatitis C virus (HCV) NS4B protein, a multispanning endoplasmic reticulum (ER) membrane protein, generates intracellular rearrangements of ER-derived membranes, essential for HCV replication. In this study, we characterized NS4B elements involved in the process of targeting, association and retention in the ER membrane. We investigated the localization and membrane association of a number of C- or N-terminal NS4B deletions expressed as GFP chimeras by biochemical and fluorescence microscopy techniques. A second set of GFP-NS4B chimeras containing the plasma membrane ecto-ATPase CD39 at the C-terminus of each NS4B deletion mutant was used to further examine the role of N-terminal NS4B sequences in ER retention. Several structural elements, besides the first two transmembrane domains (TMs), within the NS4B N-terminal half (residues 1-130) were found to mediate association of the NS4B-GFP chimeras with ER membranes. Both TM1 and TM2 are required for ER anchoring and retention but are not sufficient for ER retention. Sequences upstream of TM1 are also required. These include two putative amphipathic alpha-helices and a Leucine Rich Repeat-like motif, a sequence highly conserved in all HCV genotypes. The N-terminal 55peptidic sequence, containing the 1st amphipathic helix, mediates association of the 55N-GFP chimera with cellular membranes including the ER, but is dispensable for ER targeting of the entire NS4B molecule. Importantly, the C terminal 70peptidic sequence can associate with membranes positive for ER markers in the absence of any predicted TMs. In conclusion, HCV NS4B targeting and retention in the ER results from the concerted action of several NS4B structural elements. PMID- 20001748 TI - Endosonographic elastography of the anal sphincter in patients with fecal incontinence. AB - OBJECTIVE: In fecal incontinence the role of elastography has not yet been evaluated. We performed a trial to further characterize the internal and external anal sphincter in patients with fecal incontinence and compared a visual assessment scale with a computerized program for quantifying elastic properties of the anal sphincter. MATERIAL AND METHODS: Fifty consecutive patients with fecal incontinence were studied (n = 31 following lower anterior resection, n = 8 with Crohn's disease, n = 9 following colon surgery, n = 2 others). Elastogram color distribution within the sphincter representing elastic properties was quantified using a visual analog scale and an off-line computerized area calculation program. RESULTS: The main finding was that the inner anal sphincter (IAS) differed significantly from the external anal sphincter (EAS) with regard to elastogram color distribution. There were no significant correlations with clinical and functional parameters. There was, however, a non-significant increase in the percentage of blue (hard) areas in the IAS in patients neoadjuvantly irradiated for rectal or cervical cancer compared to non-irradiated patients, which was accompanied by a significant decrease in the resting sphincter pressure (p < 0.009). CONCLUSIONS: The IAS, a smooth muscle, and the EAS, a striated muscle, have different elastogram color distributions, probably reflecting their different elastic properties. The absence of significant correlations with the major clinical and functional parameters suggests that in routine clinical practice ultrasound real-time elastography may not yield additional information in patients with fecal incontinence. There may be exceptions, particularly in irradiated patients. PMID- 20001749 TI - Rebound acid hypersecretion from a physiological, pathophysiological and clinical viewpoint. AB - OBJECTIVE: The recent description of dyspepsia in healthy individuals after stopping treatment with proton-pump inhibitors (PPIs) indicates that reflux disease may worsen due to this treatment. The aim of this paper is to review current knowledge of the regulation of gastric acid secretion, including maximal acid secretion, and to improve understanding of the pathogenesis of acid-related conditions. MATERIAL AND METHODS: We reviewed our findings from three decades of studies on gastric acid secretion in the isolated rat stomach and in humans as well as studies by the group of Robert Jensen involving gastrinoma patients. RESULTS: The parietal cell has receptors for histamine and acetylcholine, whereas the gastrin receptor is localized to the enterochromaffin-like (ECL) cell. Gastrin-stimulated histamine release depends on the ECL cell mass, which is regulated by gastrin. The parietal cell mass is also influenced by gastrin. All conditions with hypergastrinemia concomitant with a normal oxyntic mucosa result in an increase in acid secretion. Helicobacter pylori infection in the antral mucosa may induce duodenal ulcers by its effect on acid secretion, as in patients with gastrinoma. Whereas PPIs induce clinically important rebound acid hypersecretion, histamine-2 blockers do not, since they also induce tolerance. CONCLUSION: From a biological and physiological point of view, patients should be given treatment that disturbs the normal physiology as little as possible. PMID- 20001750 TI - High-density lipoprotein cholesterol improves the model for end-stage liver disease scoring system for prognostic prediction of acute liver failure. PMID- 20001753 TI - Outcomes and safety of rapid desensitization for chemotherapy hypersensitivity. AB - IMPORTANCE OF THE FIELD: The incidence of hypersensitivity reactions (HSRs) to chemotherapy agents has increased because of increasing number of cancer survivors are exposed to repeated courses of sensitizing agents. Replacement with an alternative chemotherapy regimen is often limited by tumor sensitivity. Rapid desensitization offers an effective mean to allow continuation of the treatment to which patients have presented HSRs. AREAS COVERED IN THIS REVIEW: We review the methods, outcome and safety of the rapid desensitization protocol developed at Brigham and Women's Hospital, Harvard Medical School Affiliate, based on our recent publication "'Hypersensitivity reactions to chemotherapy: outcome and safety of rapid desensitization in 413 cases". Literature search was conducted through Medline (from January 1976 to September 2009), using PubMed. WHAT THE READER WILL GAIN: The article will give insight to clinical manifestations of immediate HSR to various chemotherapy agents and their presumably different immunopathomechanism. Risk assessment, including skin testing in those presented HSRs to platins and details on rapid desensitization process and its pitfalls will be discussed. TAKE HOME MESSAGE: Standa"rd protocol of rapid desensitization, administering under multidisciplinary team approach, is safe and effective in overcoming immediate HSRs to platins, taxanes, doxorubicin and rituximab via both intravenous and intraperitoneal routes. PMID- 20001754 TI - Inherent and iatrogenic immune defects in hairy cell leukemia: revisited. AB - IMPORTANCE OF THE FIELD: T- and NK-cell abnormalities, decreased number and function of monocytes, neutrophils and dendritic cells, decreased production of certain cytokines and increased incidence of certain autoimmune conditions have been identified in untreated hairy cell leukemia (HCL) patients. These alterations are responsible for an increased rate of infections and additional malignancies in HCL. AREAS COVERED IN THIS REVIEW: The authors offer a focused review of the most relevant preclinical and clinical studies exploring the immune abnormalities in both untreated and treated HCL. WHAT THE READER WILL GAIN: The use of potent immunosuppressive agents such as cladribine and pentostatin in HCL therapeutics has generated increasing concerns about the likelihood of additional immune impairments in these patients. While the NK cells, monocytes and neutrophils were shown to recover shortly after complete responses are achieved with these agents, the CD4(+) T-cell counts may require in excess of 2 - 3 years in order to re-enter the normal range. TAKE HOME MESSAGE: Given the advent of new molecular, genetic and immunologic techniques, a comprehensive characterization of the immune abnormalities in untreated and treated HCL represents a reachable goal and could translate into improved outcomes in clinical practice. PMID- 20001755 TI - Genetic predisposition of life-threatening antiepileptic-induced skin reactions. AB - IMPORTANCE OF THE FIELD: Recent advances in pharmacogenetic studies have uncovered increasingly more genes that predispose individuals to adverse drug reactions. Aromatic antiepileptic drugs (AEDs) are a frequent cause of severe cutaneous adverse reactions (SCAR). A strong genetic association between HLA B*1502 and carbamazepine (CBZ)-induced Stevens-Johnson syndrome (SJS) and toxic epidermal necrolysis (TEN) has been shown in Han Chinese patients. AREAS COVERED IN THIS REVIEW: This article reviews and updates genetic information associated with CBZ and other AEDs causing SCAR in different ethnic populations. WHAT THE READER WILL GAIN: Independent studies from different countries confirmed that patients carrying the HLA-B*1502 are at high risk of SJS/TEN when exposed to CBZ. The US FDA and similar regulatory agencies in Canada and Taiwan have updated the CBZ drug label to include the genetic information. Available data also suggest that HLA-B*1502 is a risk allele for SJS/TEN caused by other aromatic AEDs with a similar structure to CBZ. TAKE HOME MESSAGE: Screening for HLA-B*1502 allele before starting treatment with CBZ is justified in patients from high-risk populations as recommended by regulatory agencies. Similar chemicals should also be avoided in individuals who test positive for HLA-B*1502. PMID- 20001756 TI - Efficacy and safety of dronedarone: a review of randomized trials. AB - IMPORTANCE OF THE FIELD: Dronedarone is developed for treatment of atrial fibrillation (AF) or flutter (AFL). It is a noniodinized amiodarone analogue and believed to be without the adverse effects of amiodarone. However, long-term adverse effects are not yet well investigated. AREAS COVERED IN THIS REVIEW: This is a review of seven studies on dronedarone. WHAT THE READER WILL GAIN: DAFNE established an effective dose to be 400 mg b.i.d. ADONIS and EURIDIS showed significant prevention of AF/AFL recurrence hazard ratio (HR 0.78 and 0.73) compared to placebo. In ATHENA, cardiovascular death/hospitalization was significantly reduced (HR 0.76) in patients with AF and additional risk factors. ANDROMEDA was stopped because dronedarone increased early mortality (HR 2.13) in advanced heart failure (HF). ERATO found that dronedarone significantly reduced heart rate compared to placebo in patients with AF. DIONYSOS showed that amiodarone was superior to dronedarone to maintain sinus rhythm in patients with AF/AFL. TAKE HOME MESSAGE: Dronedarone is superior to placebo but less efficient than amiodarone in maintaining sinus rhythm in patients with a history of AF. In patients with AF and risk factors dronedarone reduces cardiovascular mortality and morbidity, but in patients with severe HF dronedarone significantly increases mortality. PMID- 20001757 TI - Clinical safety of ranibizumab in age-related macular degeneration. AB - IMPORTANCE OF THE FIELD: Clinical safety of pharmaceutical products in the elderly is vital because of their increased risk of cardiac and other adverse events. AREAS COVERED IN THIS REVIEW: Search of the Medline database, including articles and abstracts from 1984 to 2009. WHAT THE READER WILL GAIN: Knowledge of ocular and systemic risks: The rate of endophthalmitis was 0.05% per injection (MARINA) and <0.1% per injection (ANCHOR), rates confirmed in a retrospective analysis of 14,320 injections. Moderate increases in intraocular pressure were transient, and incidences of intraocular inflammation were rarely serious. Systemic arterial thromboembolic events occurred in 4.6 and 0% of ranibizumab treated patients and in 3.8 and 0% of sham-treated patients in MARINA (2 years) and PIER (1 year), respectively. In SAILOR, there was a numerically higher rate of cerebrovascular stroke with 0.5 mg ranibizumab compared with 0.3 mg ranibizumab (1.2 vs 0.7%), which was a non-statistically significant trend in patients with a history of stroke. TAKE HOME MESSAGE: Although further studies to investigate the risk of stroke with ranibizumab therapy are required, repeated intravitreal ranibizumab was well tolerated and not associated with clinically significant safety risks during up to 2 years of treatment. PMID- 20001758 TI - Survivors of acute lymphoblastic leukemia and body mass changes. AB - IMPORTANCE OF THE FIELD: Obesity is a rapidly growing challenge that has now reached epidemic proportions. Along with malnutrition, it causes increasing morbidity and mortality in the general population. Survivors of pediatric leukemia are at increased risk of developing adverse body mass changes. Despite many studies, mechanisms of regulation of fat tissue metabolism are still poorly understood. AREAS COVERED IN THIS REVIEW: The present article reviews the data from studies of leukemia survivors in the context of basic science studies and reports of nutritional situation in Europe published between 1994 and 2009. As regulation of appetite and energy balance is very complex, environmental, biochemical and genetic factors are presented. WHAT THE READER WILL GAIN: Fat mass and obesity associated gene (FTO) has recently been found to contribute to the risk of obesity. The possible role of this gene as well as late consequences of body mass changes are discussed. TAKE HOME MESSAGE: Both underweight and overweight leukemia survivors need to be monitored for ongoing health consequences of abnormal BMI. Parameters of metabolic syndrome should be included as routine assessments in outpatient clinics taking care of childhood leukemia survivors. PMID- 20001759 TI - Preoperative radiotherapy downregulates the nuclear expression of hypoxia inducible factor-1alpha in rectal cancer. AB - OBJECTIVE: To assess the value of hypoxia-inducible factor-1alpha (HIF-1alpha) expression as a predictor of disease outcome in rectal cancer treated by preoperative radio- or chemoradiotherapy. MATERIAL AND METHODS: Operative samples from 168 rectal cancer patients and 79 respective preoperative biopsies were analyzed for nuclear HIF-1alpha protein expression using immunohistochemistry by three approaches: (a) positive/negative, (b) the percentage of HIF-positive cancer cells and (c) staining intensity. The patients had received either short- (n = 75) or long-course radiotherapy with or without chemotherapy (n = 39) or no treatment preoperatively (n = 54). RESULTS: HIF-1alpha staining was positive in 70% of the diagnostic biopsies but negative in most of the post-radiotherapy specimens (60%). HIF-1alpha expression in the biopsies was downregulated in 56% of samples taken after preoperative treatment, while negative HIF-1alpha expression was upregulated in 25% of samples. Patients who had HIF-negative tumours after long-course radiotherapy had significantly (P = 0.001) better disease-specific survival (DSS) in univariate analysis. In the multivariate (Cox) regression model, HIF-1alpha lost its significance and only being in the preoperative treatment group was an independent predictor of disease-free survival. In a similar Cox model, disease recurrence and the number of metastatic lymph nodes were independent predictors of DSS. CONCLUSIONS: HIF-1alpha expression was positive in most of the preoperative biopsies but downregulated in most of the operative samples, implicating that preoperative radiotherapy downregulates HIF-1alpha expression in rectal cancer. Negative HIF expression after preoperative long-course radiotherapy was associated with significantly better DSS. PMID- 20001760 TI - Enrichment of pinolenic acid at the sn-2 position of triacylglycerol molecules through lipase-catalyzed reaction. AB - Reports have shown that Delta-5 polyunsaturated fatty acids (-5 PUFA) are enriched at sn-1,3 positions of triacylglycerols (TAG) in pine (Pinus koraiensis) nut oil (Pn). As a major Delta-5 PUFA, pinolenic acid (Pi) is about 14.2% in the oil, while the percentage of Pi at the sn-1 and/or sn-3 positions in TAG was found more than 20%. In this current study, the enhancement of Pi at the sn-2 position has been achieved by acyl migration during the lipase-catalyzed inter esterification between Pn and palm stearin (Ps). After reaction, the proportion of Pi increased at sn-2 positional fatty acid even is similar to that in total fatty acid; for example, in the inter-esterified product of 50:50 (Pn:Ps), the same amount of Pi (7.1%) present was detected both at the sn-2 and sn-1,3 positions. However, the reduction of phytosterols and tocopherols are observed in the inter-esterified products. PMID- 20001761 TI - The relationship between dietary lipids and cognitive performance in an elderly population. AB - Epidemiological studies on the association between diet and cognitive function suggested a possible role of dietary fatty acids in cognitive decline. The aim of the present study was to examine whether intake of different types of fatty acids is associated with cognitive status. A cohort of 304 (127 men and 177 women) institutionalized elderly people, with a mean age of 75.3 +/- 6.7 years, were studied. Subjects were evaluated for global cognitive functions (Mini-Mental State Examination [MMSE], Spanish version). Fatty acid intake was assessed with a semi-quantitative food frequency questionnaire. Intake of eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) were found to be predictors of cognitive impairment as they were negatively associated with the MMSE score. In accordance with this, fish intake was inversely associated with cognitive impairment. On the contrary, the n-6/n-3 polyunsaturated fatty acid ratio was positively related to the MMSE score. These results could not be explained by differences in age, sex, education, smoking behaviour, inactivity, alcohol, institution or energy intake. We suggest that consumption of EPA and DHA should be encouraged for reducing the risk of cognitive impairment and subsequently disability in elderly people. PMID- 20001762 TI - Role of oxate, phytate, tannins and cooking on iron bioavailability from foods commonly consumed in Mexico. AB - The objectives of this research were to assess the bioavailability of iron in foodstuffs found in the Mexican diet, to provide data on the content of iron absorption inhibitors present in plant origin products and to assess the inhibitory effect of these compounds and of cooking on iron bioavailability; therefore, total content and bioavailable iron, tannins, phytic and oxalic acid were determined in vegetables, cereals, legumes and animal products, before and after cooking. Vegetables, although rich in iron, have poor iron bioavailability and a high content of inhibitory factors; cooking reduced the content of iron and inhibitory factors, whereas in animal products the treatment of cooking did not significantly reduce it. Iron bioavailability, phytate content and the phytate to iron molar ratio predicted poor iron bioavailability and, therefore, a negative impact on the nutritional status of people who rely on them as staple foods could be expected. PMID- 20001744 TI - Potential therapeutic benefits of strategies directed to mitochondria. AB - The mitochondrion is the most important organelle in determining continued cell survival and cell death. Mitochondrial dysfunction leads to many human maladies, including cardiovascular diseases, neurodegenerative disease, and cancer. These mitochondria-related pathologies range from early infancy to senescence. The central premise of this review is that if mitochondrial abnormalities contribute to the pathological state, alleviating the mitochondrial dysfunction would contribute to attenuating the severity or progression of the disease. Therefore, this review will examine the role of mitochondria in the etiology and progression of several diseases and explore potential therapeutic benefits of targeting mitochondria in mitigating the disease processes. Indeed, recent advances in mitochondrial biology have led to selective targeting of drugs designed to modulate and manipulate mitochondrial function and genomics for therapeutic benefit. These approaches to treat mitochondrial dysfunction rationally could lead to selective protection of cells in different tissues and various disease states. However, most of these approaches are in their infancy. PMID- 20001763 TI - Lipoatrophy associated with the use of insulin analogues: a new case associated with the use of insulin glargine and review of the literature. AB - IMPORTANCE OF THE FIELD: Any insulin formulation can in principle cause lipoatrophy; even cases associated with recombinant human insulin have been reported. An increasing number of case reports have been published indicating that lipoatrophy also develops after treatment with various insulin analogues. AREAS COVERED IN THIS REVIEW: In this review, we summarise the literature on lipoatrophy associated with the use of insulin analogues published to date. A new case of lipoatrophy associated with the use of glargine is presented. WHAT THE READER WILL GAIN: Readers will gain insight into: i) pathogenesis of lipoatrophy associated with the use of insulin analogues and ii) clinical features of lipoatrophy. TAKE HOME MESSAGE: Twelve cases with lipoatrophy under treatment with insulin analogues have been reported so far. The exclusive occurrence in lean type 1 diabetic patients, its overlap with further autoimmune diseases and the overrepresentation of female individuals point to an immune pathogenesis. The respective exposition to the analogues lispro, aspart, glargine and detemir prior to lipoatrophy development varied considerably between 4 weeks and 2 years. No spontaneous substantial recovery of lipoatrophic areas has been reported. Frequent use of the same pen needle and lack of rotating of insulin injection sites seem to favour the development of lipoatrophy. PMID- 20001764 TI - Safety and efficacy of over-the-counter cough and cold medicines for use in children. AB - IMPORTANCE OF THE FIELD: Over-the-counter (OTC) cough and cold medications have been used widely for years and continue to be a preferred choice for temporary relief of symptoms of upper respiratory tract infections in children. These medications are being placed under extraordinary scrutiny in the pediatric population due to the lack of conclusive evidence about their therapeutic efficacy and increased reports of associations with serious adverse events and even mortality. AREAS COVERED IN THIS REVIEW: A PubMed search was conducted to identify articles published up to August 2009 describing the efficacy and safety of OTC cough and cold medications in children. The objective was to provide an overview of the relevant literature and regulatory history and to comment on the available data on this important topic. WHAT THE READER WILL GAIN: The paper provides a detailed up-to-date review of the key efficacy and safety studies published on the subject. In addition, the reader is presented with an overview of the regulatory history and recent developments surrounding the use of OTC cough and cold medications in children in the US. TAKE HOME MESSAGE: This review confirms the lack of efficacy of OTC cough and cold products in children and reaffirms that although the overall incidence of related serious adverse events is low, such events continue to occur. The conclusions in this paper support a recommendation that OTC cough and cold medications should not be given to infants and very young children. Furthermore, additional research is needed to evaluate the safety and efficacy of these medicines in the broader pediatric population. PMID- 20001765 TI - A more active treatment has profound effects on the health status of rheumatoid arthritis (RA) patients: results from a population-based RA register in Malmo, Sweden, 1997-2005. AB - OBJECTIVE: Population-based studies on the trends and effects of modern antirheumatic treatment are scarce. The aim of this study was to examine trends in treatment, health-related quality of life (HRQL), and disease outcome in a population-based register of patients with rheumatoid arthritis (RA) in Malmo, Sweden. METHODS: A continuously updated population-based RA register was established in the city of Malmo, southern Sweden, in 1997. Self-completed postal questionnaires issued in 1997, 2002, and 2005 were used to collect information on demographics, medication, and health status. Cross-sectional comparisons were made between data from 1997, 2002, and 2005. RESULTS: Between 1997 and 2005, the proportion of patients treated with any disease-modifying anti-rheumatic drug (DMARD) including biologics increased substantially (from 52% to 87%), as well as the proportion treated with methotrexate (from 23% to 52%) and biologics (almost exclusively tumour necrosis factor inhibitors) (from 0% to 20%). Twelve per cent of RA patients received biologics 5 years from disease onset in 2005. In parallel with changes in treatment, mean Health Assessment Questionnaire (HAQ) scores (1.19 vs. 0.89) and all Short Form 36 (SF-36) subscales improved from 1997 to 2005 (non-overlapping confidence intervals). CONCLUSION: Between 1997 and 2005, there was a substantial increase in the use of DMARDs, which was accompanied by improved mean HAQ and SF-36 scores in cross-sectional comparisons. These results support the concept that more intensive treatment with DMARDs and biologics can have profound effects on the overall health status in RA patients at the population level. PMID- 20001766 TI - Bone and joint involvement in Fabry disease. AB - Fabry disease (FD) is an X-linked lysosomal storage disease caused by deficient activity of the enzyme alpha-galactosidase A. Although the disease has progressive effects on most organ systems in the body, data is limited regarding skeletal involvement in this rare disorder. We describe four family-related patients, three men and one premenopausal female, sharing a classic phenotype of FD. Dual-energy X-ray was performed in all cases and osteoporosis or osteopenia were found in all patients and osteoporotic fractures in one. One patient also showed both neuropathic joint disease and osteonecrosis. Several mechanisms that may explain osteoporosis and osteoarthropathy in the setting of FD are emphasized. PMID- 20001767 TI - Expression of IL-10 family cytokines in rheumatoid arthritis: elevated levels of IL-19 in the joints. AB - OBJECTIVES: Interleukin (IL)-10 functions as an anti-inflammatory cytokine in rheumatoid arthritis (RA). New IL-10 family cytokines IL-19, IL-20, IL-22, IL-24, and IL-26 have recently been discovered. Information concerning the expression and function of these cytokines in autoimmune diseases is currently limited. The aim of this study was to investigate their expression in RA. METHODS: mRNA levels of the cytokines were studied using quantitative reverse transcriptase polymerase chain reaction (RT-PCR). Peripheral blood (PB) and synovial fluid (SF) mononuclear cells (MCs), purified T cells, and monocytes/macrophages from RA patients and healthy volunteers, and synovial tissues from patients with RA or osteoarthritis (OA), were examined. The expression of IL-19 protein in T cells and monocytes/macrophages was studied by flow cytometry. RESULTS: IL-10 and IL-19 mRNA levels were significantly elevated in SFMCs from patients with RA compared with PBMCs from RA patients or healthy volunteers. IL-20 and IL-22 mRNA levels were also upregulated in RA SFMCs but their level of expression was lower than that of IL-10 or IL-19. Importantly, synovial tissue IL-19 levels in RA were increased when compared with OA. IL-19 expression was upregulated in both T cells and macrophages derived from patients with RA. IL-1beta increased IL-19 levels in PBMCs, suggesting that elevated levels of IL-1 in RA joints may contribute to upregulated IL-19 expression. CONCLUSIONS: The majority of the IL-10 family cytokines are expressed in RA. IL-19 demonstrated the highest expression in rheumatoid joints, and could thus be involved in the regulation of synovial inflammation in RA. PMID- 20001769 TI - Proposal would increase member dues in 2011. PMID- 20001770 TI - Is it time for limited licensure? PMID- 20001771 TI - What is your diagnosis? Transitional cell carcinoma in the bladder of a dog. PMID- 20001772 TI - Pathology in practice. Systemic cryptococcosis caused by C. neoformans and concomitant severe pulmonary aelurostrongylosis. PMID- 20001773 TI - A federal and state transport plan for movement of eggs and egg products from commercial egg production premises in a high-pathogenicity avian influenza control area. PMID- 20001776 TI - Echocardiographic evaluation of dogs with dysautonomia. AB - OBJECTIVE: To describe echocardiographic findings in dogs with dysautonomia. DESIGN: Prospective case series: ANIMALS: 20 dogs with dysautonomia (13 confirmed during necropsy and 7 with results of antemortem testing [tear production, pilocarpine response test, atropine response test, and ID histamine response] supportive of the diagnosis). PROCEDURES: Dogs with dysautonomia were evaluated by use of echocardiography, and M-mode measurements were obtained on all dogs. A dobutamine response test was performed on 1 dog, starting at a rate of 1 microg/kg/min and doubling the rate every 15 minutes until fractional shortening (FS) increased to > 2 times the baseline value. RESULTS: Evidence of systolic dysfunction was detected in 17 of 20 dogs with dysautonomia, as determined on the basis of FS (median, 17.9%; range, 4.0% to 31.1%). Left ventricular internal dimension during diastole or left ventricular internal dimension during systole was enlarged in 4 of 20 and 14 of 20 dogs, respectively. Enlargement of the left atrium or aorta was identified in 3 of 15 and 1 of 15 dogs in which it was measured, respectively. Administration of dobutamine at a rate of 4 microg/kg/min resulted in dramatic improvement in FS (increase from 4% to 17%) in the 1 dog tested. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggested that echocardiographic evidence of diminished systolic function was common in dogs with dysautonomia. Whether the diminished function was a result of sympathetic denervation or myocardial hibernation was unclear, although myocardial hibernation was more likely. PMID- 20001778 TI - Prevalence and prognostic impact of hypocobalaminemia in dogs with lymphoma. AB - OBJECTIVE: To determine the prevalence of hypocobalaminemia in dogs with multicentric lymphoma and to investigate any relationship between serum cobalamin concentration and disease outcome. DESIGN: Cohort study. ANIMALS: 58 dogs with multicentric lymphoma. PROCEDURES: Serum cobalamin concentrations were measured in 58 dogs with multicentric lymphoma. Clinical signs, stage, and immunophenotype for dogs with hypocobalaminemia were compared with those for dogs with serum cobalamin concentrations above the lower end of the reference range. Survival times for dogs undergoing a cyclic multidrug chemotherapy protocol (n = 53) were similarly compared. Serum cobalamin concentrations for treated dogs that died or were euthanized before day 60 were compared with those of dogs still alive at day 60. RESULTS: Serum cobalamin concentrations ranged from < 150 to 1,813 ng/L, with a median concentration of 401 ng/L. Nine of the 58 (16%) dogs had hypocobalaminemia (serum cobalamin concentration < 252 ng/L). Three of 9 dogs with hypocobalaminemia survived to at least day 60, compared with 40 of 44 (91%) dogs without hypocobalaminemia (serum cobalamin concentration >or= 252 ng/L). Ten (10/53 [19%]) dogs undergoing a cyclic multidrug chemotherapy protocol died before day 60, and the median serum cobalamin concentration for these dogs (232 ng/L) was significantly lower than for those still alive at the end point of the study (556 ng/L). CONCLUSIONS AND CLINICAL RELEVANCE: Hypocobalaminemia was relatively uncommon in this population of dogs with multicentric lymphoma, but was associated with a poor outcome. Serum cobalamin concentrations may provide prognostic information in dogs with multicentric lymphoma. PMID- 20001779 TI - Pregabalin as an adjunct to phenobarbital, potassium bromide, or a combination of phenobarbital and potassium bromide for treatment of dogs with suspected idiopathic epilepsy. AB - OBJECTIVE: To assess tolerability and short-term efficacy of oral administration of pregabalin as an adjunct to phenobarbital, potassium bromide, or a combination of phenobarbital and potassium bromide for treatment of dogs with poorly controlled suspected idiopathic epilepsy. DESIGN: Open-label, noncomparative clinical trial. ANIMALS: 11 client-owned dogs suspected of having idiopathic epilepsy that was inadequately controlled with phenobarbital, potassium bromide, or a combination of these 2 drugs. PROCEDURES: Dogs were treated with pregabalin (3 to 4 mg/kg [1.4 to 1.8 mg/lb], PO, q 8 h) for 3 months. Number of generalized seizures in the 3 months before and after initiation of pregabalin treatment was recorded. Number of responders (>or= 50% reduction in seizure frequency) was recorded, and seizure frequency before and after initiation of pregabalin treatment was compared by use of a nonparametric Wilcoxon signed rank test. RESULTS: Seizures were significantly reduced (mean, 57%; median, 50%) after pregabalin administration in the 9 dogs that completed the study; 7 were considered responders with mean and median seizure reductions of 64% and 58%, respectively. Adverse effects for pregabalin were reported in 10 dogs. Mean and median plasma pregabalin concentrations for all dogs were 6.4 and 7.3 microg/mL, respectively. CONCLUSIONS AND CLINICAL RELEVANCE: Pregabalin may hold promise as a safe and effective adjunct anticonvulsant drug for epileptic dogs poorly controlled with the standard drugs phenobarbital or potassium bromide. Adverse effects of pregabalin appeared to be mild. Additional studies with larger numbers of dogs and longer follow-up intervals are warranted. PMID- 20001780 TI - Rate and frequency of recurrence of uroliths after an initial ammonium urate, calcium oxalate, or struvite urolith in cats. AB - OBJECTIVE: To determine frequency of and interval until recurrence after initial ammonium urate, calcium oxalate, and struvite uroliths in cats and whether breed, age, or sex was associated with increased risk for urolith recurrence. DESIGN: Case-control study. ANIMALS: 4,435 cats with recurrent uroliths. PROCEDURES: To identify recurrence of uroliths in cats for which uroliths were submitted for analysis at the Minnesota Urolith Center in 1998, the facility's database was searched for urolith resubmissions from the same cats between 1998 and 2003. Risk factors and differences in mean interval until recurrence were assessed. RESULTS: Of 221 cats with ammonium urate uroliths in 1998, 29 (13.1%) had a first and 9 (4.1%) had a second recurrence. Mean interval until recurrence was 22 and 43 months for the first and second recurrence, respectively. Of 2,393 cats with calcium oxalate uroliths in 1998, 169 (7.1%) had a first, 15 (0.6%) had a second, and 2 (0.1%) had a third recurrence. Mean interval until recurrence was 25, 38, and 48 months for the first, second, and third recurrence, respectively. Of 1,821 cats with struvite uroliths in 1998, 49 (2.7%) had a first and 3 (0.2%) had a second recurrence. Mean interval until recurrence was 29 months for first and 40 months for second recurrences. CONCLUSIONS AND CLINICAL RELEVANCE: These results provided insights into the frequency of urolith recurrence in cats. Because some uroliths associated with recurrent episodes probably were not submitted to our facility, our data likely represented an underestimation of the actual recurrence rate. PMID- 20001781 TI - Echocardiographic and clinicopathologic characterization of pericardial effusion in dogs: 107 cases (1985-2006). AB - OBJECTIVE: To evaluate sensitivity and specificity of echocardiography for diagnosis of cardiac masses in dogs with pericardial effusion. DESIGN: Retrospective case series. ANIMALS: 107 dogs with pericardial effusion. PROCEDURES: Records of dogs with pericardial effusion examined at the University of California-Davis Veterinary Medical Teaching Hospital from 1985 to 2006 were reviewed. Dogs were included when echocardiography and pericardectomy or necropsy were performed. Sensitivity, specificity, and metastatic rates were calculated for various causes of pericardial effusion. RESULTS: 107 dogs with pericardial effusion were evaluated by surgery (n = 48 dogs), necropsy (44), or both (15). Echocardiography revealed no mass (n = 41 dogs), a right atrial (RA) mass (38), a heart base (HB) mass (23), a pericardial mass (2), an HB and an RA mass (2), and a right ventricular mass (1). Sensitivity and specificity were 82% and 100%, respectively, for detection of a cardiac mass; 82% and 99%, respectively, for detection of an RA mass; and 74% and 98%, respectively, for detection of an HB mass. Most HB masses were neuroendocrine or ectopic thyroid gland tissue, but 3 were hemangiosarcomas and 4 were mesotheliomas. Most RA masses were hemangiosarcomas, but this group also included a neuroendocrine tumor, ectopic thyroid gland tissue, mesothelioma, lymphosarcoma, and sarcoma. Metastatic rates did not differ (50% to 66%) among neoplastic causes. CONCLUSIONS AND CLINICAL RELEVANCE: Echocardiography had high sensitivity and specificity for diagnosis and differentiation of RA or HB masses in dogs with pericardial effusion. There was a high rate of metastasis for cardiac masses of all causes. PMID- 20001782 TI - Autoimmune myasthenia gravis in a ferret. AB - CASE DESCRIPTION: A 7-month-old neutered male ferret was evaluated for episodic pelvic limb weakness of 2 weeks' duration. CLINICAL FINDINGS: Neurologic examination revealed flaccid tetraparesis with decreased spinal reflexes suggestive of a neuromuscular disease. Results of hematologic and CSF analyses, thoracic radiography, and abdominal ultrasonography were unremarkable. Electrodiagnostic testing revealed subtle spontaneous activity localized to pelvic limb interosseous muscles, unremarkable motor nerve conduction velocities, and lower than typical compound muscle action potential (CMAP) amplitude for tibial nerve stimulation only. A severe decremental response of the CMAP was detected with repetitive nerve stimulation (45.5% at the third ulnar nerve). An esophagogram revealed mild megaesophagus. Intravenous neostigmine methylsulfate administration resulted in immediate resolution of muscle weakness. Cross reacting anti-acetylcholine receptor (AChR) antibodies were detected in serum (0.35 nmol/L) by use of a canine- and feline-specific muscle extract. Clinical signs and ancillary test results were diagnostic of acquired myasthenia gravis. TREATMENT AND OUTCOME: Pyridostigmine bromide was administered (1 mg/kg [0.45 mg/lb], PO, q 8 h), resulting in complete remission of clinical signs. However, 1 month after the diagnosis, the ferret was euthanized because of recurrence of weakness despite anticholinesterase treatment. CLINICAL RELEVANCE: To the authors' knowledge, this is the first report of acquired myasthenia gravis in a ferret and the first identification of anti-AChR antibodies in this species. Autoimmune myasthenia gravis should be considered in ferrets when weakness and flaccid paresis suggest a neuromuscular disease. Electrodiagnostic testing, anticholinesterase challenge, and AChR antibody titer determination were helpful for diagnosis of this condition. PMID- 20001783 TI - Successful treatment of pacemaker-induced stricture and thrombosis of the cranial vena cava in two dogs by use of anticoagulants and balloon venoplasty. AB - CASE DESCRIPTION: 2 castrated male Labrador Retrievers (dogs 1 and 2) were evaluated 3 to 4 years after placement of a permanent pacemaker. Dog 1 was evaluated because of a large volume of chylous pleural effusion. Dog 2 was admitted for elective replacement of a pacemaker. CLINICAL FINDINGS: Dog 1 had mild facial swelling and a rapidly recurring pleural effusion. Previously detected third-degree atrioventricular block had resolved. Cranial vena cava (CVC) syndrome secondary to pacemaker-induced thrombosis and stricture of the CVC was diagnosed on the basis of results of ultrasonography, computed tomography, and venous angiography. Dog 2 had persistent third-degree atrioventricular block. Intraluminal caval stricture and thrombosis were diagnosed at the time of pacemaker replacement. Radiographic evidence of pleural effusion consistent with CVC syndrome also was detected at that time. TREATMENT AND OUTCOME: Dog 1 improved after treatment with unfractionated heparin and a local infusion of recombinant tissue-plasminogen activator. Balloon venoplasty was performed subsequently to relieve the persistent caval stricture. In dog 2, balloon dilatation of the caval stricture was necessary to allow for placement of a new pacing lead. Long-term anticoagulant treatment was initiated in both dogs. Long term (> 6 months) resolution of clinical signs was achieved in both dogs. CLINICAL RELEVANCE: Thrombosis and stricture of the CVC are possible complications of a permanent pacemaker in dogs. Findings suggested that balloon venoplasty and anticoagulation administration with or without thrombolytic treatment can be effective in the treatment of dogs with pacemaker-induced CVC syndrome. PMID- 20001784 TI - Systemic hypertension in a dog with a functional thyroid gland adenocarcinoma. AB - CASE DESCRIPTION: A 12-year-old 21.9-kg (48.48-lb) spayed female Alaskan Malamute with a long-term history of panting, polydypsia and polyuria, weight loss, hind limb weakness, and a decrease in appetite was evaluated for hypertension. CLINICAL FINDINGS: Use of Doppler sphygmomanometry revealed a systolic blood pressure of 250 mm Hg (mean value for 5 consecutive measurements). Palpation of the ventral cervical region revealed a fixed asymmetric mass in the area of the lobes of the thyroid gland. The portion of the mass on the right side was approximately 2 x 2 cm, whereas the portion of the mass on the left side was approximately 1 x 1.5 cm. Hyperthyroidism was diagnosed on the basis of high serum thyroxine concentrations. Thyroidectomy of both lobes of the gland was performed. Histologic examination revealed a bilateral, multilobulated, and encapsulated thyroid gland adenocarcinoma. TREATMENT AND OUTCOME: Thyroidectomy of both lobes of the gland was performed with clinical resolution of hypertension. The dog was treated postoperatively with chemotherapeutics, including doxorubicin and carboplatin, and external beam radiation. Calcium and thyroxine homeostasis fluctuated; however, the dog finally achieved concentrations within the respective reference ranges through the administration of calcitriol, calcium carbonate, and levothyroxine. CLINICAL RELEVANCE: This report describes concurrent hypertension in a dog with a functional thyroid gland adenocarcinoma with subsequent return of blood pressure values to within reference ranges after thyroidectomy. PMID- 20001785 TI - Acute generalized exanthematous pustulosis with lymphangitis triggered by a spider bite. AB - Acute generalized exanthematous pustulosis (AGEP) is a rare, severe cutaneous reaction pattern that, in the majority (>90%) of cases, is related to administration of medication. It can be seen in both genders and in all ages. The cutaneous manifestations of AGEP are usually seen 1-14 days after drug administration. A 39-year-old woman presented to our outpatient clinic with the complaint of generalized erythema, burning, and rash. She explained that 2 days before presentation a spider bite had occurred on her left forearm, after which she had experienced pain and erythema spreading gradually to the left upper extremity. On her dermatologic examination, she had an indurated necrotic plaque on the left forearm, which had an upward-spreading linear erythema. Additionally, she had diffuse erythema on her body and small pustules over erythematous skin, especially located on the left popliteal fossa and gluteal region. Based on the clinical and histopathologic findings, she was diagnosed as having AGEP. Because there was no drug use in her history, we attributed her AGEP lesions to the spider bite. This case is interesting, because the patient also had lymphangitis. Herein, we present the fifth case reported in the literature of AGEP caused by a spider bite. PMID- 20001786 TI - Using ImageJ for the quantitative analysis of flow-based adhesion assays in real time under physiologic flow conditions. AB - This article intends to close the gap between the abundance of regular articles focusing on adhesive mechanisms of cells in a flow field and purely technical reports confined to the description of newly developed algorithms, not yet ready to be used by users without programming skills. A simple and robust method is presented for analysing raw videomicroscopic data of flow-based adhesion assays using the freely available public domain software ImageJ. We describe in detail the image processing routines used to rapidly and reliably evaluate the number of adherent and translocating platelets in videomicroscopic recordings. The depicted procedures were exemplified by analysing platelet interaction with immobilized von Willebrand factor and fibrinogen in flowing blood under physiological wall shear rates. Neutralizing GPIbalpha function reduced shear-dependent platelet translocation on von Willebrand factor and abolished firm platelet adhesion. Abciximab, Tirofiban and Eptifibatide completely inhibited GPIIb/IIIa-dependent stable platelet deposition on fibrinogen. The presented method to analyse videomicroscopic recordings from flow-based adhesion assays offers the advantage of providing a simple and reliable way to quantify flow-based adhesion assays, which is completely based on ImageJ and can easily be applied to study adhesion mechanisms of cells in non-fluorescent modes without the need to deviate from the presented protocol. PMID- 20001787 TI - Expression of EspA in Lactococcus lactis NZ9000 and the detection of its immune effect in vivo and vitro. AB - Enterhemorrhagic Escherichia coli (EPEC), an important cause of severe infantile diarrheal disease in many parts of the developing world, produced several recently described virulence determinations. Several of its virulence factors are secreted by type III secretion including EspA, which forms filamentous structures on bacterial surface bridging to the host cell's surface. These structures on bacterial surfaces may deliver other virulence factors directly into the host cell from EHEC. In this study, EspA was expressed in Lactococcus lactis NZ9000 (L. lactis NZ9000). BALB/C mice were immunized by recombinant EspA, and mice sera were assayed for the disruption of E. coli O157:H7 interaction with the host cell. BALB/C mice which were immunized with recombinant EspA produced specific antibody titers, and the difference between the control group and the immune group is marked (P<0.05). The polyclonal mice antisera blocked E. coli O157:H7 induced host cell actin rearrangement and could label E. coli O157:H7 EspA filaments in vitro. Our present findings suggest that L. lactis may be a good candidate to produce oral vaccine, and thus entertains the possibility of developing EspA-oral vaccine in the defense of E. coli O157:H7 infection. PMID- 20001788 TI - Effects of IL-1 molecules on growth patterns of 3-MCA-induced cell lines: an interplay between immunogenicity and invasive potential. AB - The balance between inflammation and immunity is cardinal for the outcome of the malignant process. Local attenuated inflammatory responses mediated by innate cells may provide accessory signals for the development of acquired immunity against malignant cells. In contrast, excessive inflammatory responses accompany tumorigenesis and tumor invasiveness, by the induction of immunosuppression. In the present study, we have assessed the role of tumor cell-derived IL-1 in determining the invasive versus immunostimulatory potential of tumor cells. For this purpose, we have used 3-MCA-induced fibrosarcoma cell lines from IL-1 knockout (KO) versus control mice. Cell lines with no IL-1 failed to establish tumors in intact mice, while lines obtained from control mice were invasive and induced a potent angiogenic response. In contrast, cell lines from IL-1KO mice were more immunogenic. SDF-1 and IL-6, each induced by IL-1, were the two major cytokines whose levels differed in cell lines with or without IL-1. We could not detect differences in cell surface markers related to immunogenicity, such as MHC Class I, co-stimulatory, or adhesion molecules between both types of cells. However, more T-cells were observed at the inoculation site of tumor cells devoid of IL-1 and more pronounced parameters related to anti-tumor immunity were observed in the spleen (IL-12 and IFNgamma) of these mice, compared to mice bearing tumors derived from control mice, where host-derived IL-1 is present. In addition, injection of tumor cells devoid of IL-1, which failed to grow in mice, induced an anti-tumor cell immune memory, while in mice injected with tumor cells from control mice; no immune memory could be detected. From the results, it seems that IL-1 is a crucial factor in determining the balance between immunity and inflammation in tumor-bearing mice. This suggests that manipulation of IL-1 could be useful in anti-tumor therapy, by reducing invasiveness and promoting immunity against the malignant cells. PMID- 20001789 TI - Effect of needle diameter and flow rate on rat and human mesenchymal stromal cell characterization and viability. AB - INTRODUCTION: Current mesenchymal stromal cell (MSC) delivery methods require infusion/implantation through needles and/or catheters. Little investigation into the effect of delivery via catheter injection has been completed. We hypothesize that injection of rat and human MSCs through various clinically relevant-sized catheters and flow rates will not affect cell viability, characterization, or function. METHODS: Both rat and human MSCs were injected through 20-, 25-, and 30 gauge needles, as well through an SL-10 microcatheter at rates of 60, 120, 240, and 500 mL/h. MSC viability and apoptotic fraction was measured. MSCs were characterized 24 h after injection with flow cytometric immunophenotyping, and multilineage differentiation was completed. RESULTS: Catheter diameter or flow rate did not affect rat MSC viability. No clinically significant decrease in human MSC viability was observed immediately after injection; however, a delayed decrease in viability was observed at 24 h. No difference in the surface markers CD11b, CD45, CD29, CD49e, CD73, CD90, CD105, and Stro-1 or the capacity for multilineage differentiation (adipogenesis, osteogenesis, and chondrogenesis) was observed for either rat or human MSCs. CONCLUSION: The injection of human and rat MSCs through various clinically relevant catheters and flow rates did not have a clinically significant effect on viability immediately after injection, indicating compliance with recently published Food and Drug Administration guidelines (viability >70%). Further, no changes in cell characterization or function were observed via measurement of cell surface markers and the capacity for multilineage differentiation, respectively. These results ensure the biocompatibility of MSCs with commonly used delivery methods. PMID- 20001790 TI - Current role of breast duct microendoscopy in malignancy. PMID- 20001791 TI - Potential use of ultrasound for the detection of cell changes in cancer treatment. PMID- 20001793 TI - Dissecting out the role of the CCK-2 receptor in the carcinogenic effects of progastrin in the colon. PMID- 20001794 TI - Hot issues in the current management of eye cancer. PMID- 20001795 TI - Updated lung cancer staging system. AB - The tumor, node and metastasis classification of malignant tumors is periodically revised. Its seventh edition includes the updated classification for lung cancer, based on the analyses of the International Association for the Study of Lung Cancer international database. It is the largest validation ever carried out to date: 100,869 patients registered in 46 databases from 20 countries. The analysis of this database allowed a detailed study of the impact of tumor size on prognosis, the reclassification of additional tumor nodules, the reclassification of pleural dissemination and the separation of two groups of metastatic disease. These changes led to modifications in stage grouping that better differentiate tumors with different prognosis. This updated classification is also recommended for small-cell lung cancer, and for broncho-pulmonary carcinoids. A new international nodal map with stations and zones, and a histological definition of visceral pleura invasion, have also been proposed. PMID- 20001797 TI - Prognostic value of nodal ratios in node-positive breast cancer: a compiled update. AB - The number of positive axillary nodes is a strong prognostic factor in breast cancer, but is affected by variability in nodal staging technique yielding varying numbers of excised nodes. The nodal ratio of positive to excised nodes is an alternative that could address this variability. Our 2006 review found that the nodal ratio consistently outperformed the number of positive nodes, providing strong arguments for the use of nodal ratios in breast cancer staging and management. New evidence has continued to accrue confirming the prognostic significance of nodal ratios in various worldwide population settings. This review provides an updated summary of available data, and discusses the potential application of the nodal ratio to breast cancer staging and prognostication, its role in the context of modern surgical techniques such as sentinel node biopsy, and its potential correlations with new biologic markers such as circulating tumor cells and breast cancer stem cells. PMID- 20001796 TI - Critical review of prostate cancer predictive tools. AB - Prostate cancer is a very complex disease, and the decision-making process requires the clinician to balance clinical benefits, life expectancy, comorbidities and potential treatment-related side effects. Accurate prediction of clinical outcomes may help in the difficult process of making decisions related to prostate cancer. In this review, we discuss attributes of predictive tools and systematically review those available for prostate cancer. Types of tools include probability formulas, look-up and propensity scoring tables, risk class stratification prediction tools, classification and regression tree analysis, nomograms and artificial neural networks. Criteria to evaluate tools include discrimination, calibration, generalizability, level of complexity, decision analysis and ability to account for competing risks and conditional probabilities. The available predictive tools and their features, with a focus on nomograms, are described. While some tools are well-calibrated, few have been externally validated or directly compared with other tools. In addition, the clinical consequences of applying predictive tools need thorough assessment. Nevertheless, predictive tools can facilitate medical decision-making by showing patients tailored predictions of their outcomes with various alternatives. Additionally, accurate tools may improve clinical trial design. PMID- 20001798 TI - Prognostic role of tumor size in childhood cancer. AB - Identifying prognostic factors that influence outcome is a key point in pediatric oncology, as it enables risk stratification and helps us to fine-tune risk adapted treatments, thereby hopefully improving the chances of survival for high risk patients, and minimizing the burden of treatment for those whose outlook is good. Along with other clinical and biological variables, tumor size at onset has been identified as a crucial variable in several pediatric malignancies, and in soft-tissue sarcomas in particular. This review provides a critical reappraisal of the prognostic role of tumor size in pediatric soft-tissue sarcomas. PMID- 20001799 TI - Epigenetics of neurological cancers. AB - Epigenetic mechanisms involving DNA methylation, histone modifications and noncoding RNAs regulate and maintain gene-expression states. Similar to genetic mutations, alterations in epigenetic regulation can lead to uncontrolled cell division, tumor initiation and growth, invasiveness and metastasis. Research in brain cancer, particularly gliomas, has uncovered global and gene-specific DNA hypomethylation, local DNA hypermethylation of gene promoters and the de regulation of microRNA expression. Understanding epigenetic dysregulation in brain cancers has provided new tools for prognostication, as well as suggesting new approaches to therapy. There is significant interest in new sequencing-based technologies that map genetic and epigenetic alterations comprehensively and at high resolution. These methods are being applied to brain tumors, and will better define the contribution of epigenetic defects to tumorigenesis. PMID- 20001802 TI - Roles of LPA in ovarian cancer development and progression. AB - Lysophosphatidic acid (LPA), a bioactive phospholipid, stimulates survival, proliferation, adhesion, migration and invasion of ovarian cancer cells through the activation of G-protein-coupled plasma membrane receptors. LPA and its receptors are aberrantly expressed in ovarian cancer, with high levels predominantly found in malignant ascites and in the plasma of ovarian cancer patients. LPA signals multiple intracellular pathways, such as Ras/MEKK1-MAPK and PI3K/Akt, to promote growth factors and protease expression, and induce angiogenesis and tumor cell invasion through the extracellular matrix and across the basement membrane. Only a small portion of this intricate lipid-signaling cascade has been characterized thus far. We believe that elucidation of this complex transduction network will provide further opportunities to understand the mechanism of ovarian carcinogenesis, invasion and metastasis. PMID- 20001800 TI - Muc4/MUC4 functions and regulation in cancer. AB - The membrane mucin MUC4 (human) is abundantly expressed in many epithelia, where it is proposed to play a protective role, and is overexpressed in some epithelial tumors. Studies on the rat homologue, Muc4, indicate that it acts through anti adhesive or signaling mechanisms. In particular, Muc4/MUC4 can serve as a ligand/modulator of the receptor tyrosine kinase ErbB2, regulating its phosphorylation and the phosphorylation of its partner ErbB3, with or without the involvement of the ErbB3 ligand neuregulin. Muc4/MUC4 can also modulate cell apoptosis via multiple mechanisms, both ErbB2 dependent and independent. Muc4/MUC4 expression is regulated by multiple mechanisms, ranging from transcriptional to post-translational. The roles of MUC4 in tumors suggest that it may be valuable as a tumor marker or target for therapy. PMID- 20001801 TI - Pathogenesis of ovarian cancer: clues from selected overexpressed genes. AB - Ovarian cancer is the most malignant gynecologic neoplasm. Although new chemotherapeutic agents have improved patients' 5-year survival rate, the overall mortality of ovarian cancer has remained largely unchanged in the past several decades. The main reason for the lack of success in effectively treating ovarian cancer is our limited understanding of its etiology and the very few molecular diagnostic markers and therapeutic targets known so far. Identification and characterization of ovarian cancer-associated genes are fundamental for unveiling the pathogenesis of its initiation and progression, especially the development of recurrent diseases. As there are a vast number of genes for which molecular genetic changes and aberrant gene expression have been reported in ovarian cancer, this review will only focus on summarizing those exemplified genes that have been demonstrated to have biological functions in promoting ovarian cancer development and potential clinical significance. The genes to be discussed include nuclear proteins (Notch3, HBXAP [Rsf-1], NAC1 and NFkappaB), cytoplasmic proteins (fatty acid synthase and apolipoprotein E) and cell surface/secretory proteins (mucin-4, mesothelin, claudin, HLA-G, kallikrein and folate receptor and osteopontin). Since the study of ovarian cancer-associated genes is complicated by several factors unique to ovarian cancer, we will also present our views on the limitations and challenges of current ovarian cancer research. PMID- 20001803 TI - Targeted therapy in biliary tract cancer: 2009 update. AB - Biliary tract cancers (BTCs) include cholangiocarcinoma (intrahepatic, perihilar and extrahepatic), carcinoma of the gall bladder and ampullary carcinoma. In patients with advanced disease the prognosis is poor. There is not a consensus regarding treatment strategy. Chemotherapy has only limited efficacy. This review summarizes the new approaches for BTC patients and the rationale for targeted therapies. The prognostic factors and the molecular features of BTC are analyzed. The clinical trials evaluating the targeted agents are accurately described, especially those assessing the role of anti-EGFR and antiangiogenic drugs. The ongoing trials are also analyzed. In fact, only the results of these trials will establish which is the most effective agent or combination for this setting. PMID- 20001806 TI - Art, surgery and transplantation. AB - Roy Calne (1930-) has elegantly cultivated the science and art of transplantation. Throughout his medical and artistic career his eyes have remained fully open not only to science and scientific advances but to any new developments that would enhance his art of painting as applied to his patients, colleagues, events and surgical operations related to transplantation. Calne contributed to and developed a new field through art in the understanding of the lives of his patients, the working of his colleagues and the application of surgical principles to the specialty in which he labours, surgical transplantation. The application of "Art, Surgery and Transplantation" should in many ways be the fountain of information and support for those seeking this way of therapy. PMID- 20001807 TI - Surgical history. AB - The importance of surgical history for surgeons and students of this field of knowledge is undeniable. Knowing surgical history makes us aware of findings and discoveries we never knew before. Surgical history helps us in defining how surgery evolved through the centuries and mostly how those observations have shaped surgical thought. In this writing, I introduce my own observations of the field and explain how I grew to learn and appreciate the evolution of surgical history through my own personal experience. And, as important, at least for me, I narrate my observations as to how my education and contributions to the discipline shaped my knowledge of surgical history. PMID- 20001804 TI - New hopes from old drugs: revisiting DNA-binding small molecules as anticancer agents. AB - Most of the anticancer chemotherapeutic drugs that are broadly and successfully used today are DNA-damaging agents. Targeting of DNA has been proven to cause relatively potent and selective destruction of tumor cells. However, the clinical potential of DNA-damaging agents is limited by the adverse side effects and increased risk of secondary cancers that are consequences of the agents' genotoxicity. In this review, we present evidence that those agents capable of targeting DNA without inducing DNA damage would not be limited in these ways, and may be as potent as DNA-damaging agents in the killing of tumor cells. We use as an example literature data and our own research of the well-known antimalarial drug quinacrine, which binds to DNA without inducing DNA damage, yet modulates a number of cellular pathways that impact tumor cell survival. PMID- 20001808 TI - Comparing FK-506 with basic fibroblast growth factor (b-FGF) on the repair of a peripheral nerve defect using an autogenous vein bridge model. AB - OBJECTIVE: The limited availability of donor sites for nerve grafts and the morbidity associated with their harvesting serve as motivating factors to actively conduct research to find alternatives to the status quo. Experimental and clinical studies have shown that a vein segment used to bridge a peripheral nerve defect leads to a functional nerve repair. Both FK-506 and b-FGF have been reported to enhance peripheral nerve regeneration. This study compared the effects of FK-506 with that of b-FGF on peripheral nerve regeneration in a rat autogenous vein graft conduit model. METHODS: The main trunk of the right sciatic nerve was transected and bridged by an autogenous vein in 30 rats. Small osmotic pumps were placed just proximal to the anastomoses. Groups of 10 rats were assigned to receive saline solution, b-FGF (2,000 units), or FK-506 (0.5 mg/kg/day) via the osmotic pumps for 2 weeks. Sciatic nerve regeneration was evaluated by sensory function, walking track analysis, electrophysiologic studies, and light microscopic evaluation. RESULTS: On post-operative day 90, there was a statistically significant difference (p <.005) in nerve regeneration between the rats who received saline compared with those who received FK-506 or b FGF. This was determined using sensory function tests, sciatic function index, and electrophysiologic studies. The number of nerve axons, as determined by histological analysis, revealed there were significantly more nerve fibers which were regenerated in both experimental groups (FK-506 and b-FGF) when compared with rats who received saline. There was no statistically significant difference in the number of nerve axons that regenerated in rats injected with FK-506 vs. rats injected with b-FGF. CONCLUSION: FK-506 and b-FGF promote similar nerve regeneration in rats compared with control. PMID- 20001809 TI - Retrograde arterial flush of the liver graft in living donor liver transplantation. AB - Formal hepatic arterial flush to preserve the liver graft in living donor liver transplantation (LDLT) is not recommended by most transplant centers because direct cannulation may injure the intima of the hepatic artery. The authors describe a method of retrograde arterial flush of the liver graft without arterial cannulation by hepatic venous outflow occlusion (HVOO) in LDLT. First, we proved no backflow of the hepatic artery without HVOO by portal flush to pig livers. Then we used HVOO on 15 LDLT cases (Group HVOO). The results were compared with those of 24 counterpart LDLT cases (Group non-HVOO) without hepatic artery flush. The two-week posttransplantational liver functions were not different between two groups except that the day-three and day-seven serum bilirubin levels were lower in Group HVOO (day-three total bilirubin: 4.99 +/- 4.04 mg/dl versus 7.65 +/- 4.33 mg/dl, p =.016; day-seven total bilirubin: 5.06 +/- 5.02 mg/dl versus 9.57 +/- 6.09, p =.005). The rates of vascular complications, six-month graft survival, and biliary anastomotic stricture in Group HVOO were 0, 93.3, and 13.3% respectively, which were not different from those of Group non-HVOO. In summary, to avoid intima injury, the retrograde arterial flush of liver graft by HVOO is safe in LDLT. The short-term results showed the effect of decreasing early functional cholestasis but the long-term benefits need further evaluation, especially with regards to biliary anastomotic complications. PMID- 20001810 TI - Effects of CO(2) pneumoperitoneum on anastomotic healing in rats receiving preoperative 5-fluorouracil neoadjuvant chemotherapy. AB - BACKGROUND: When used separately, antineoplastic agents and carbon dioxide (CO(2)) pneumoperitoneum have been reported to impair anastomotic healing in experimental animals. However, the effects of their combined use have not been previously investigated. The aim of this study was to investigate the possibility that neoadjuvant chemotherapy with 5-fluorouracil followed by CO(2) pneumoperitoneum would affect the healing of anastomoses in the colon. METHODS: Sprague-Dawley rats (n = 48) were given 5-fluorouracil (20 mg/kg/day) for 5 days, and were then assigned to one of the three groups. Prior to surgery, the control group received no pneumoperitoneum. The other two groups received pneumoperitoneum at 6 and 12 mmHg, respectively, for 2 hr. The large intestine was transected and anastomosis was performed via median laparotomy. On postoperative days 3 and 7, relaparotomy was performed in half of the rats in each group. From the colon, a segment including the anastomosis was excised. Tissue hydroxyproline levels were measured. For histological evaluation, the Verhofstad scale was modified and used. RESULTS: No significant differences in hydroxyproline levels were seen across the groups on postoperative days 3 or 7. However, by postoperative day 7, polymorphonuclear leukocytes and necrosis in the 6-mmHg group had decreased markedly, and granulation had improved. CONCLUSION: Overall, these findings suggest that preoperative 5-fluorouracil therapy followed by pneumoperitoneum at 6 or 12 mmHg does not impair anastomotic healing. PMID- 20001811 TI - Serum neopterin as a prognostic indicator in patients with gastric carcinoma. AB - PURPOSE: To evaluate the possible association between serum neopterin level and clinical and laboratory findings and their prognostic value of patients with gastric carcinoma. METHODS: Serum samples were collected from patients (n = 38) and normal volunteers (n = 39) and stored at -80 degrees C until analyzed. Neopterin, C-reactive protein (CRP), and IL-6 concentration were measured by ELISA. The clinicopathological parameters were determined by reviewing both medical charts and pathological records. RESULTS: Mean neopterin levels were 15.26 +/- 11.46 nmol/L in patients with gastric carcinoma and 9.87 +/- 2.90 nmol/L in the control group. Serum neopterin concentrations were significantly higher in patients with gastric carcinoma than in the control group. The number of patients with elevated neopterin level was significantly correlated with stage, gastric wall involvement, the number of metastatic lymph nodes, and the level of serum CRP. The mean neopterin concentrations were significantly elevated in patients older than 60 years, in the presence of venous invasion and increased metastatic lymph node number, and in patients with elevated CRP levels. Neopterin level was also correlated with overall survival as an independent prognostic indoicator. CONCLUSIONS: Serum neopterin levels were elevated in patients with advanced gastric cancer and correlated with prognostic parameters and overall survival. Moreover, neopterin measured at the time of diagnosis can be used to predict the survival of gastric carcinoma. PMID- 20001812 TI - Amyand's hernia in elderly patients: diagnostic, anesthetic, and perioperative considerations. AB - The presence of a vermiform appendix in an inguinal hernia sac is termed as Amyand's hernia. Although rare, mistakes in diagnosis and treatment can cause catastrophic results. Charts of patients with inguinal hernia were reviewed, and four cases of Amyand's hernia were confirmed. The clinical presentation, anesthetic, and perioperative management of Amyand's hernia were further analyzed. The mean age of patients was over 70 years, and all were males. None of the patients were diagnosed preoperatively. All the patients had little abdominal complaint only with a right inguinal mass and dragging sensation for several hours. Due to the short time after incarceration and significant cardiovascular and pulmonary comorbidities, manual reduction was attempted first in three patients. With complete preoperative evaluation and careful perioperative support, all patients underwent appendectomy and Bassini's hernia repair through a groin incision. Based on age-related organ failure and associated chronic medical illnesses of geriatric patients, the difficulties in the diagnosis and treatment are also summarized and analyzed. PMID- 20001813 TI - Valve system does not reduce the catheter-related bloodstream infection. AB - BACKGROUND: The Groshong catheter (GC) is considered to have a lower risk of central venous catheter-related bloodstream infection (CVC-RBSI) than conventional catheters because of its valve system (closed-end) for preventing blood reflux. However, few studies have compared the GC with conventional (open end) catheters in terms of catheter-related complications. PURPOSE: To compare the incidence of catheter-related complications including CVC-RBSI between the GC and the Argyle catheter (AC). METHODS: The GC and the AC were inserted in the same way from the internal jugular vein. Catheter-related complications were evaluated from the database retrospectively. RESULTS: Two hundred seventy GCs were inserted in 123 patients, and 251 ACs were inserted in 135 patients. There were no significant differences in patient background factors between GC and AC use, except for the following two parameters. Use of GC was associated with a longer catheter insertion length and a younger patient age. Univariate analysis revealed that neither type of catheter reduced the incidence of CVC-RBSI. Kaplan Meier analysis and log rank test revealed no significant difference between the GC and the AC in the period from insertion to development of complications. CONCLUSIONS: The GC has no superiority over the conventional AC for preventing CVC-RBSI. PMID- 20001814 TI - Morphological changes during creation of a neo-bile duct using a vein and a biodegradable endoluminal stent. AB - Major bile duct lesions are usually treated by a hepaticojejunostomy which is often complicated by cholangitis and liver fibrosis. The aim of this study was to investigate the morphologic features of a neo-bile duct created from a vein and a biodegradable endoluminal stent. The neo-bile duct was created using a segment of the external jugular vein which was endoluminally stented by a biodegradable poly lactate-acid stent. In 18 pigs, the common bile duct was resected and replaced by the vein with (n = 12) or without endoluminal stent (n = 6). Six animals served as controls. Survival, liver function and morphological changes of the neo-bile duct and the liver were observed for six months. After six months, the neo-bile duct morphologically resembled the native bile duct showing Ck7-positive columnar epithelium and newly formed capillaries in the bile duct wall. The biodegradable stent disappeared after four months. All animals survived and showed normal liver function and no cholestasis. In contrast, after sole vein reconstruction of the bile duct, four animals died due to biliary peritonitis and cholangitis. Creation of a neo-bile duct which morphologically resembles the native bile duct is feasible by using a body's own vein and a biodegradable endoluminal stent. PMID- 20001815 TI - Guidelines for rodent survival surgery. PMID- 20001816 TI - Principles of anesthesia monitoring-capnography. PMID- 20001818 TI - Confronting a neglected epidemic: tobacco cessation for persons with mental illnesses and substance abuse problems. AB - Tobacco use exerts a huge toll on persons with mental illnesses and substance abuse disorders, accounting for 200,000 of the annual 443,000 annual tobacco related deaths in the United States. Persons with chronic mental illness die 25 years earlier than the general population does, and smoking is the major contributor to that premature mortality. This population consumes 44% of all cigarettes, reflecting very high prevalence rates plus heavy smoking by users. The pattern reflects a combination of biological, psychosocial, cultural, and tobacco industry-related factors. Although provider and patient perspectives are changing, smoking has been a historically accepted part of behavioral health settings. Additional harm results from the economic burden imposed by purchasing cigarettes and enduring the stigma attached to smoking. Tailored treatment for this population involves standard cessation treatments including counseling, medications, and telephone quitlines. Further progress depends on clinician and patient education, expanded access to treatment, and the resolution of existing knowledge gaps. PMID- 20001819 TI - Overview of the symposium on public health significance of genomics and eco genetics. AB - Genomic and genetic information is rapidly becoming a major element in public health research and emerging public health practice. This symposium reviews the methods, findings, and significance of genome-wide association studies from epidemiological and statistical points of view. We examine infectious and inflammatory components of gene-environment interaction in the respiratory system. We note the need for nutrient and dietary data and many other kinds of environmental exposure data in population-based genomic studies. Then we explore the sufficiency of a well-informed family history for public health and family counseling purposes. Finally, in an era of direct-to-consumer genomic test promotion, we review the evidence on the critical question, will genetic risk profiles motivate individuals and families to choose more healthful behaviors? This symposium builds on the foundation of the symposium on Public Health Genetics in Volume 21 (2000) of the Annual Review of Public Health. PMID- 20001820 TI - Outcome-based workforce development and education in public health. AB - The broad scope of the public health mission leads to an increasingly diverse workforce. Given the range of feeder disciplines and the reality that much of the workforce does not have formal training in public health science and practice, a pressing need exists for training and education throughout the workforce. Just as we in public health take a rigorous approach to our science, so too should we take a rigorous, evidence-driven approach to workforce development. In this review, we recommend a framework for workforce education in public health, integrating three critical conceptual approaches: (a) adult learning theory; (b) competency-based education; and (c) the expanded Dreyfus model in public health, an addition to the Dreyfus model of professional skills progression. We illustrate the application of this framework in practice, using the field of applied epidemiology. This framework provides a context for designing and developing high-quality, outcome-based workforce development efforts and evaluating their impact, with implications for academic and public health practice efforts to educate the public health workforce. PMID- 20001822 TI - Bioengineered three-dimensional physiological model of colonic longitudinal smooth muscle in vitro. AB - BACKGROUND: The objective of this study was to develop a physiological model of longitudinal smooth muscle tissue from isolated longitudinal smooth muscle cells arranged in the longitudinal axis. METHODS: Longitudinal smooth muscle cells from rabbit sigmoid colon were isolated and expanded in culture. Cells were seeded at high densities onto laminin-coated Sylgard surfaces with defined wavy microtopographies. A highly aligned cell sheet was formed, to which addition of fibrin resulted in delamination. RESULTS: (1) Acetylcholine (ACh) induced a dose dependent, rapid, and sustained force generation. (2) Absence of extracellular calcium attenuated the magnitude and sustainability of ACh-induced force by 50% and 60%, respectively. (3) Vasoactive intestinal peptide also attenuated the magnitude and sustainability of ACh-induced force by 40% and 60%, respectively. These data were similar to force generated by longitudinal tissue. (4) Bioengineered constructs also maintained smooth muscle phenotype and calcium dependence characteristics. SUMMARY: This is a novel physiologically relevant in vitro three-dimensional model of colonic longitudinal smooth muscle tissue. Bioengineered three-dimensional longitudinal smooth muscle presents the ability to generate force, and respond to contractile agonists and relaxant peptides similar to native longitudinal tissue. This model has potential applications to investigate the underlying pathophysiology of dysfunctional colonic motility. It also presents as a readily implantable band-aid colonic longitudinal muscle tissue. PMID- 20001823 TI - Evaluation of quality of life in people with aphasia using a Dutch version of the SAQOL-39. AB - PURPOSE: To examine the quality of life (QoL) of people with aphasia and to study the influence of variables such as age, time post onset and (degree of) social support on the QoL of aphasic persons. METHOD: We compared the scores of an aphasic population (N = 43) with those of a healthy control group (N = 43) and of a group of patients with brain lesions without neurogenic communication disorders (N = 43) on a Dutch version of the Stroke and Aphasia Quality of Life-scale (SAQoL-39) and on a social support questionnaire. In half of the aphasic group, the SAQoL-39 was re-administered 8 months after the first testing. RESULTS: People with aphasia obtained significantly lower scores for QoL measures compared with both other groups. Especially, communicative and psychosocial factors seem to influence these results. Older people with aphasia scored lower than younger persons and women tend to evaluate their QoL somewhat more negatively than men. Persons who had aphasia for more than 6 months tended to have higher QoL-scores compared with those who had become aphasic more recently. After 8 months, the retested group scored significantly higher on communication and on psychosocial functioning than on first testing. CONCLUSIONS: Gathering information on QoL after suffering from stroke and from aphasia can lead to a better understanding of the problems involved. The clinical use of instruments such as the SAQoL-39 can probably contribute to a more patient oriented rehabilitation, whereby the focus not only lie in improving linguistic skills but also on reducing the impairments and the handicaps that accompany aphasia and thus on increasing QoL. PMID- 20001821 TI - Integrating clinical, community, and policy perspectives on human papillomavirus vaccination. AB - Infection with genital human papillomavirus (HPV) may cause anogenital cancers, oropharyngeal cancers, anogenital warts, and respiratory papillomas. Two prophylactic vaccines (a bivalent and a quadrivalent vaccine) are now licensed and currently in use in a number of countries. Both vaccines prevent infection with HPV-16 and HPV-18, which together cause approximately 70% of cervical cancers, and clinical trials have demonstrated 90%-100% efficacy in preventing precancerous cervical lesions attributable to HPV-16 and HPV-18. One vaccine also prevents HPV-6 and HPV-11, which cause 90% of genital warts. A growing literature describes psychosocial, interpersonal, organizational, and societal factors that influence HPV vaccination acceptability. This review summarizes the current literature and presents an integrated perspective, taking into account these diverse influences. The resulting integrated framework can be used as a heuristic tool for organizing factors at multiple levels to guide intervention development and future research. PMID- 20001824 TI - A Delphi study on environmental factors that impact work and social life participation of individuals with multiple sclerosis in Austria and Switzerland. AB - PURPOSE: This study aimed to gain knowledge about environmental factors (EFs) that impact work and social life participation of people with multiple sclerosis (MS) in Austria and Switzerland to extend the knowledge of participation and to identify key areas for measuring participation. METHOD: A three-round Delphi study was conducted defining patients as experts. In the 1st round, qualitative data was gathered through questionnaires, analyzed with content analysis, and factors were assigned to EFs as classified in the ICF. In the 2nd and 3rd round, experts judged EFs according to its relevance to obtain consensus (cut-off 75%). Categories were ranked on a scale from mostly important to important. RESULTS: One hundred and twelve Austrian and 109 Swiss experts were recruited. The content analysis revealed 768 EFs. The study resulted in a list of 176 consensus factors for Austria and 177 Switzerland. Five categories revealed to be highly important, 12 moderately important, 6 fairly important, and 10 important. CONCLUSIONS: This study indicates that participation in work or social life is influenced by physical, social, attitudinal, and policy factors. Consensus factors afford insights into areas for consideration in the development of participation outcome measurements and support a comprehensive and inclusive rehabilitation approach. PMID- 20001825 TI - Being in- or out-of-sync: couples' adaptation to change in multiple sclerosis. AB - PURPOSE: To examine how couples adapt to the challenges of multiple sclerosis (MS) and to identify possible risk factors for relational stress. METHOD: Semi structured interviews were conducted with eight couples to explore how participants defined and identified the strengths and challenges in their relationship, negotiated role changes and received external support. Conceptual frameworks on how families adapt to chronic illness guided within and across case analyses. We identified patterns in the couples' current responses to the demands and stressors of MS. RESULTS: Two patterns of adaptation to MS were identified as being 'in-sync' or 'out-of-sync'. Characteristics of the four couples currently in-sync included having a relapsing-remitting type of MS, which proceeded at a pace that enabled both partners to maintain their social roles and identity, and a collaborative problem solving style. The four couples currently out-of-sync had a rapid progression of MS, loss of employment before retirement age, differences in personal styles that shifted from being complementary to oppositional in the face of increased demands and struggles with parenting adolescent children. CONCLUSIONS: Clinicians can use these data to assess possible relational strain in couples with MS and identify families who might benefit from referrals to family therapy or other relational support. PMID- 20001826 TI - Absence of combined effects of anemia and bilateral surgical status on inpatient rehabilitation outcomes following total knee arthroplasty. AB - PURPOSE: To examine the effects of anemia on inpatient rehabilitation outcomes following unilateral and bilateral total knee arthroplasty (TKA). METHOD: This was a multicenter, retrospective study from 15 inpatient rehabilitation facilities. Patients (N = 5421) with very low hematocrit (Hct <30%), low Hct (30 36% women, 30-41% men), or normal Hct (>36% women, 41% men) were included. Inpatient rehabilitation occurred following TKA. Functional independence measure (FIM) scores, length of stay (LOS), itemized hospital charges, discharge destination were main outcomes. RESULTS: LOS was 13% longer and hospital charges were 12.5-18.0% higher in the very low Hct than remaining groups (p = 0.0001). The FIM score and subscores for walking, stair climbing, bathing, transfers, and dressing changes were comparable for all Hct groups for the overall score. Hct <30% did not correspond to worse outcomes in patients with bilateral surgeries compared with unilateral surgeries; total FIM scores improved 47-53% across all Hct groups, regardless of bilateral surgical status. Discharge to home ranged 92.6-94.7% across all Hct groups (p > 0.05). CONCLUSIONS: Rehabilitation teams can expect comparable functional improvements and discharge to home in anemic and non-anemic patients with either unilateral or bilateral surgeries without hematologic correction in the rehabilitation setting, but might need an additional day to accomplish these outcomes. PMID- 20001827 TI - Usefulness of perceived level of exertion in patients with chronic low back pain attending a physical training programme. AB - PURPOSE: Firstly, to examine whether heart rate (HR) can be predicted based on the Borg-scale for perceived exertion in patients with chronic low back pain (CLBP) and secondly, to assess changes in HR, Borg-scores and workload to study the relevance of a stepwise increase of workload based on the Borg-score and HR. METHODS: Ninety-nine patients with CLBP and a mean disability-score (Roland Morris Disability Questionnaire) of 13.8 (SD = 3.7) participated in a 10-week aerobic training programme (20 min, 3 times/week). HR, ratings of perceived exertion (RPE), and workload were monitored. Prior to treatment, patients completed questionnaires on pain, disability, and several psychological factors (catastrophizing, kinesiophobia, and depression). RESULTS: The original Borg equation (i.e. 'HR = Borg x 10') appears accurate in predicting HR on a group level. Pain-related and psychological factors were not significantly related to the accuracy of the Borg equation. Whereas mean workload increased significantly (from 93.8 (SD = 29.9) to 129.5 W (SD = 39.7), p < 0.001) throughout the training programme, HR increased only slightly (from 130.2 (SD = 13.5) to 139.2 (SD = 13.9) bpm; p < 0.001) and Borg-scores remained stable (from 13.2 (SD = 1.9) to 13.3 (SD = 1.8); p = 0.48). CONCLUSIONS: The capability of the Borg-scale to accurately predict HR appears moderate. However combined with measuring HR, it results in a relevant and efficient method for training CLBP patients in clinical practice. PMID- 20001828 TI - Community services for young adults with motor disabilities - a paradox. AB - PURPOSE: To understand the contribution of educational, employment, transportation, and assured income service programs to the successful transition to adulthood of young persons with motor disabilities. METHOD: Personal interviews of 76 young adults ages 20 to 30 years with a diagnosis of cerebral palsy (n = 54) or spina bifida (n = 22) varying in functional mobility. Content analysis of transcribed interviews was conducted and themes identified. Results. 'Paradox of Services' emerged as a dominant theme across all four service sectors. Services designed to encourage independence and full participation for the young adults in their communities often restricted their independence and employment options in young adulthood. CONCLUSIONS: Services need to be more individualized and flexible to accommodate the environmental and personal needs of the young adults. Rehabilitation professionals have important roles at both an individual family level and the systems level to ensure the 'best fit' between persons with motor disabilities and the services available to them. Contemporary views of disability support rehabilitation intervention targeting both individual and environmental factors. PMID- 20001829 TI - The 'risk inventory for persons with spinal cord injury': development and preliminary validation of a risk assessment tool for spinal cord injury. AB - PURPOSE: To present a new short instrument to measure perceived risks of common everyday activities engaged in by persons with spinal cord injury (SCI), and to provide preliminary data on its psychometric properties. METHOD: Community dwelling men and women with SCI (N = 139) in metropolitan Detroit completed the risk inventory for persons with spinal cord injury (RISCI). They also answered a risk-taking identity question ('Are you a risk-taker'?) and completed the risk orientation questionnaire (ROQ) (Rohrmann, http://www.rohrmannresearch.net/ , 2008), a risk propensity measure. RESULTS: All items of the RISCI correlated positively with each other and the total score; internal reliability as measured by Cronbach's alpha was 0.86. Principal components factor analysis confirmed a one-factor structure which explained 41% of the variance. A three-factor solution with readily interpretable factors explained 64% of the variance. Content validity was established through extensive consultations with persons with SCI in the development of the measure. Discriminant validity was supported by the ability of the RISCI to distinguish between subsamples (for example, between men and women, those with paraplegia and tetraplegia) for whom differences in risk assessment might be expected. Criterion validity was supported by significant relationships in the expected directions between the RISCI and risk-taking identity and between the RISCI and the ROQ. CONCLUSIONS: Findings suggest that the RISCI is a brief, easy to administer and psychometrically sound measure of perceived risk of activities common in daily life for use with persons with SCI. PMID- 20001830 TI - Effectiveness of a community-based low intensity exercise programme for ambulatory stroke survivors. AB - PURPOSE: To establish the feasibility and effectiveness of a community-based exercise programme for ambulatory patients with stroke discharged from rehabilitation. METHOD: Eighteen participants were recruited 3-12 months after onset of first stroke. Using a time series experimental design, the group completed a baseline period of 4 weeks (A1), a group exercise programme of low intensity progressive resistive exercise and functional tasks for lower limb muscles (B) and repeat assessment after cessation of exercise (A2). Fitness instructors delivered sessions at Leisure Centres twice weekly for 14 weeks with physiotherapy support and the minimum attendance requirement was 16 sessions. Measures included muscle strength, gait velocity, Berg Balance Scale and Nottingham Extended Activities of Daily Living. RESULTS: Lower limb muscle strength improved after training (ANOVA, p < 0.02). Paretic knee extension strength increased from 43.4 + or - 5.9 to 60.4 + or - 6.8 Nm after 16 exercise sessions. Walking velocity increased significantly (ANOVA, p < 0.001), from 0.54 + or - 0.07 to 0.75 + or - 0.08 m/s (t = -3.31, p < 0.01). Balance and everyday function were also significantly improved (p < 0.003). There were marked individual variation in the response to training, and those who completed additional training did not show benefit. CONCLUSIONS: This community-based exercise programme was feasible and delivered positive improvements in physical function for participants. Further issues raised for investigation include the individual response to training and the benefits of extended training. PMID- 20001831 TI - Parents' perspectives on occupational therapy and physical therapy goals for children with cerebral palsy. AB - Contemporary rehabilitation literature emphasises functional goals for children with disabilities and use of a collaborative goal-setting process grounded in principles of family centred service delivery. PURPOSE: To explore parents' experiences with goals and goal setting. METHOD: We conducted a qualitative study with 11 focus groups and two individual interviews with 39 parents of children with cerebral palsy living in western Canada. We used an inductive, thematic analysis to identify prominent themes. RESULTS: The analysis revealed five themes representing goals that were meaningful to parents and provided insight into parents' experiences with goal-setting processes in occupational and physical therapy: (1) movement as the means to functional success; (2) physical health and fitness are important therapy goals; (3) the importance of leading happy, fulfilling lives and being accepted by others; (4) 'We can't do it all': balancing therapy with the demands of everyday life; and (5) shifting roles and responsibilities in goal setting. CONCLUSIONS: The variability noted both in parents' desired role in goal setting and in goals important to parents highlights the importance of establishing trusting relationships with families so that family goals, values, individual circumstances, and desired level of participation in goal setting can be openly discussed. PMID- 20001832 TI - Development and preclinical evaluation of acellular collagen scaffolding and autologous artificial connective tissue in the regeneration of oral mucosa wounds. AB - This work assessed wound healing response in rabbit oral lesions grafted with autologous artificial connective tissue or acellular collagen scaffolds. Autologous artificial oral connective tissue (AACT) was produced using rabbit fibroblasts and collagen I scaffolds. Before implantation, AACT grafts were assayed to demonstrate the presence of fibroblasts and extracellular matrix components, as well as the expression of characteristic genes and secretion of chemokines, cytokines, and growth factors. AACT grafts were tested in the rabbits from which the fibroblasts were obtained, whereas acellular collagen type I scaffolds (CS) were evaluated in a separate group of rabbits. In both cases, contralateral wounds closed by secondary intention were used as controls. In a separate experiment, AACT-grafted wounds were directly compared with contralateral CS-grafted wounds in the same animals. Wound contraction and histological parameters were examined to evaluate closure differences between the treatments in the three animal experiments performed. Contraction of wounds grafted with AACT and CS was significantly lower than in their controls (p < 0.05). Additionally, AACT significantly lowered wound contraction when compared with CS (p < 0.05). Intriguingly, it was observed that AACT-grafted wounds initially displayed a significantly higher (p < 0.05)-albeit transient inflammatory response than seen in CS-grafted wounds and secondary healed wounds. This suggests that an early inflammatory component may contribute to tissue regeneration. Altogether, the results suggest that AACT- and CS-grafted wounds favor regeneration of oral mucosa. PMID- 20001833 TI - The eternal closure of the biased mind? The clinical and scientific relevance of biophysics, infinitesimal dilutions, and the memory of water. PMID- 20001834 TI - What do "we" want and need to know about prayer and healing? PMID- 20001835 TI - Localization of acupuncture points BL25 and BL26 using computed tomography. AB - OBJECTIVES: The aim of this study was to provide a metric description of acupuncture points BL25 and BL26, to investigate their relation to individual anatomical landmarks and structures, and to identify anatomical structures that are involved in needle manipulation during de qi. DESIGN: Fifty-eight (58) white patients with low-back pain received 107 needlings that were documented using computed tomography (CT). OUTCOME MEASURES: For each patient, the body-mass index (BMI) and the interscapular distance were measured. Using the CT scan, the following parameters were assessed: thickness of the soft tissue layer, distance of the puncture site and the needle tip to the vertebral line, distance of the needle tip to the body surface and the intermuscular space, and the needle depth in muscle tissue. RESULTS: The mean distance from the puncture site to the vertebral line was 3.5 +/- 0.5 cm for BL25 and 3.4 +/- 0.4 cm for BL26. The distance of the needle tip to the vertebral line was similar (3.4 +/- 0.5 cm for BL25 and 3.2 +/- 0.4 cm for BL26). The mean distance of the needle tip to the body surface was 4.1 +/- 0.9 cm at BL25 and 3.9 +/- 0.8 cm at BL26. The majority of the needle tips were located in the intermuscular region between the erector spinae and the transversospinal muscles. There was a significant correlation between the interscapular distance and the thickness of the soft tissue layer with the BMI at both acupuncture points. Nearly all parameters correlated with the BMI. At BL26, correlations between the distance of needle to the vertebral line and the BMI were not statistically significant but the scatter plots indicate a positive dependency. CONCLUSIONS: The results of this study show an association between de qi and needle location within the intermuscular septa. BL25 and BL26 are located as clusters in and around the intermuscular region of the erector spinae and the transversospinal muscles, with a distance to the vertebral line of 3.49 +/- 0.58 cm and 3.32 +/- 0.53 cm, respectively. Using proportional methods is relevant for the success of acupuncture therapy. PMID- 20001836 TI - Clinical effect of a topical herbal ointment on pain in temporomandibular disorders: a randomized placebo-controlled trial. AB - OBJECTIVES: The aim of this study was to compare the effectiveness of using Ping On ointment and using petroleum jelly in the treatment of temporomandibular joint (TMJ) and masticatory muscle pain, in order to establish the true efficacy of Ping On ointment. METHODS: In this randomized, double-blinded, placebo-controlled trial, 55 subjects with TMJ and/or masticatory pain (Group I patients according to the Research Diagnostic Criteria for Temporomandibular Disorder (RDC/TMD) received Ping On ointment for 4 weeks, or placebo for 4 weeks. Subjects were evaluated with standard measures of efficacy: pain intensity measured by visual analogue scale and maximal comfortable mandibular opening, at baseline and again after 4 weeks of treatment. RESULTS: Ping On ointment significantly reduced the symptoms of painful TMJs and/or masticatory muscles. Maximal comfortable mandibular opening also improved in the Ping On ointment group compared with the placebo, but was not clinically significant. CONCLUSIONS: This preliminary study suggests that topical application of Ping On ointment may be considered for further investigation as a potential first-line treatment modality, before prescribing analgesics, for the management of TMDs. It is topically applied, safe, reversible, and effective in managing TMDs and masticatory muscle pain. PMID- 20001837 TI - Effects of Thai traditional massage on autistic children's behavior. AB - OBJECTIVES: The objective of this study was to access whether there were any therapeutic effects of Thai Traditional Massage (TTM) on major behavioral and emotional disturbances in Thai autistic children. DESIGN: This was a randomized controlled trial study. SETTINGS/LOCATION: The study was conducted at the Rehabilitation Centre of the Thai Red Cross Society. SUBJECTS: A total of 60 autistic children between the ages of 3 and 10 completed this study. INTERVENTIONS: Standard sensory integration therapy (SI) was compared to the SI with TTM treatments. OUTCOME MEASURES: Parents and teachers assessed major behavior disturbances using the Conners' Rating Scales at 0 and 8 weeks. Sleep Diary (SD), recorded by the parents, assessed the patient's sleeping patterns every week. RESULTS: Sixty (60) autistic children, mean age 4.67 +/- 1.82, were recruited. No statistical differences were seen in the demographic and baseline data among both groups. From both the Conners' Teacher Questionnaire and SD, statistical improvement was detected for conduct problem, hyperactivity, inattention-passivity, hyperactivity index, and sleeping behavior. However, results from the Conners' Parent Questionnaire revealed an improvement only for anxiety (p = 0.04) in the massage group, whereas when both groups were compared, a significant improvement in conduct problem (p = 0.03) and anxiety (p = 0.01) was found. Results indicated that TTM may have a positive effect in improving stereotypical behaviors in autistic children. CONCLUSIONS: Over a period of 8 weeks, our findings suggested that TTM could be used as a complementary therapy for autistic children in Thailand. PMID- 20001838 TI - Treating spider phobia using Neuro Emotional Technique: findings from a pilot study. AB - BACKGROUND: Specific phobia, the most common anxiety disorder, can disrupt lives, limit work efficiency, reduce self-esteem, and strain relationships. Current interventions show some degree of success, yet relapse is common. Consequently, the need for a more effective and durable intervention is evident. The purpose of this pilot study is to investigate the efficacy of a new intervention, Neuro Emotional Technique (NET), on individuals with spider phobia, and to determine whether further investigation is warranted. METHODS: Participants who met the inclusion criterion that spider phobia impacted their daily lives were randomized to either a control group that received no intervention (N = 7), or to an experimental group that received two 30-minute sessions of NET approximately 2 weeks apart (N = 8). The primary measure was the Subjective Units of Distress Scale, and secondary measures were the Spider Questionnaire, Behavioral Assessment Test, Beck Anxiety Inventory, and change in heart rate (HR). RESULTS: Compared with the no-intervention control group, statistical analysis indicates a significant advantage for the NET group in regard to state anxiety/subjective distress, reported fear, and avoidant behavior. The difference between the two groups for general anxious symptomatology (trait anxiety) and change in HR was not statistically significant. No adverse reactions were reported. CONCLUSIONS: The findings of this pilot study suggest that NET is a promising intervention for spider phobia in adults. A larger, full-scale study is required to confirm these results. PMID- 20001839 TI - Successful treatment of paraquat poisoning by Xuebijing, an injection concocted from multiple Chinese medicinal herbs: a case report. AB - BACKGROUND AND OBJECTIVES: 1,1'-Dimethyl-4,4'-bipyridinium dichloride (Paraquat) poisoning remains a significant global health problem. Despite noteworthy research and clinical efforts worldwide in the last few decades, little improvement has been made in reducing fatality from Paraquat poisoning with conventional treatment strategies. We herein report a case of successful management of Paraquat poisoning by Xuebijing, a newly developed injection concocted from multiple traditional Chinese medicinal herbs. SETTINGS/LOCATION, SUBJECTS, AND INTERVENTIONS: A 25-year-old male patient was brought to the Emergency Department at the First Affiliated Hospital of Jilin University in Changchun, China approximately 23 hours after ingestion of approximately 50 mL of 20% (w/v) Paraquat in a suicide attempt. On admission, the patient presented with clinical symptoms as well as significantly abnormal results in the liver and kidney function tests that were typical of severe Paraquat poisoning. Following the routine emergency procedures for pesticide poisoning to minimize the poison exposure and relieve the poisoning symptoms, an intravenous drip of Xuebijing together with dexamethasone was given daily to the patient until discharge 10 days after admission. The patient was followed up for 15 months after discharge, during which monthly chest radiography was performed. Treatment outcomes: At the time of discharge, the patient had recovered well: His symptoms of poisoning in the mouth and gastrointestinal tract were all diminishing; his liver and kidney functions were recovering with the major test parameters improving; his chest radiograph was clear, showing no signs of pulmonary fibrosis. During the postdischarge follow-up period, the monthly chest radiographs were all normal. CONCLUSIONS: This case report suggests that Xuebijing has great potential as a novel effective alternative to the conventional management of Paraquat poisoning. This potential needs to be further evaluated in a substantially larger number of clinical cases in the future. PMID- 20001843 TI - Chemokine profile of human serum from whole blood: migratory effects of CXCL-10 and CXCL-11 on human mesenchymal stem cells. AB - Autologous human serum is used in cartilage repair and may exert its effect by the recruitment of mesenchymal stem and progenitor cells (MSC). Aim of our study was to analyze the chemokine profile of human serum and to verify chemotactic activity of selected chemokines on MSC. Human MSC were isolated from iliac crest bone marrow aspirates. Chemotactic activity of human serum made from whole blood and pharma grade serum was tested in 96-well chemotaxis assays and chemokine levels were analyzed using human chemokine antibody membrane arrays. The chemotactic potential of selected chemokines on MSC was tested dose dependently using chemotaxis assays. Human serum derived from whole blood significantly attracted human MSC, while pharma grade serum did not recruit MSC. Human chemokine antibody array analysis showed that the level of chemokines CXCL-3, 5, 7-8, 10-12, 16; CCL- 2, 5, 11, 13, 16-20, 24-25, 27; as well as XCL-1 was elevated (fold change >1.5) in serum derived from whole blood compared to nonrecruiting pharma grade serum. Chemotaxis assays showed that the chemokines IP 10/CXCL-10 and I-TAC/CXCL-11 significantly recruit human MSC. PARC/CCL-18, HCC 4/CCL-16, CTACK/CCL-27, and Lymphotactin/XCL-1 showed no chemotactic effect on MSC. Therefore, human serum derived from whole blood contains chemokines that may contribute to serum-mediated recruitment of human mesenchymal progenitors from bone marrow. PMID- 20001844 TI - Compressive force induces osteoclast differentiation via prostaglandin E(2) production in MC3T3-E1 cells. AB - In orthodontic tooth movement, prostaglandin E(2) (PGE(2)) released from osteoblasts can alter the normal process of bone remodeling. We previously showed that compressive force (CF) controls bone formation by stimulating the production of PGE(2) and Ep2 and/or Ep4 receptors in osteoblasts. The present study was undertaken to examine the effect of CF on the production of PGE(2), cyclooxygenase-2 (COX-2), macrophage colony-stimulating factor (M-CSF), receptor activator of NF-kappaB ligand (RANKL), and osteoprotegerin (OPG) using osteoblastic MC3T3-E1 cells and to examine the indirect effect of CF on osteoclast differentiation using RAW264.7 cells as osteoclast precursors. MC3T3 E1 cells were cultured with or without continuous CF (1.0 or 3.0 g/cm(2)) for 24 hr, and PGE(2) production was determined using ELISA. The expression of COX-2, M CSF, RANKL, and OPG genes and proteins was determined using real-time PCR and ELISA, respectively. Osteoclast differentiation was estimated using tartrate resistant acid phosphatase (TRAP) staining of RAW 264.7 cells cultured for 10 days with conditioned medium from CF-treated MC3T3-E1 cells and soluble RANKL. As CF increased, PGE(2) production and the expression of COX-2, M-CSF, and RANKL increased, whereas OPG expression decreased. The number of TRAP-positive cells increased as CF increased. Celecoxib, a specific inhibitor of COX-2, blocked the stimulatory effect of CF on TRAP staining and the production of PGE(2), M-CSF, RANKL, and OPG. These results suggest that CF induces osteoclast differentiation by increasing M-CSF production and decreasing OPG production via PGE(2) in osteoblasts. PMID- 20001845 TI - Blood serum levels of amino-terminal pro-C-type natriuretic peptide in patients with systemic sclerosis. AB - The aim of this pilot study was to examine the association of serum levels of amino-terminal fragment of pro-C-type natriuretic peptide (NT-proCNP), as a substitute measure of serum C-type natriuretic peptide (CNP), with clinical and laboratory findings in patients with systemic sclerosis (SSc). Serum NT-proCNP, soluble (s)E-and sP-selectin levels were examined using specific enzyme-linked immunosorbent assay in 30 patients with SSc and 30 healthy controls. We found no statistically significant difference in serum levels of NT-proCNP neither between patients with SSc and healthy controls nor between those treated with immunosuppressive agents and untreated patients. Nevertheless, in the investigated SSc group, serum NT-proCNP levels correlated with the concentrations of C-reactive protein (CRP) and the duration of the disease. Both sP- and sE selectin levels were elevated in SSc patients when compared to healthy subjects. Also they did not correlate with the concentrations of NT-proCNP. The results of the study indicate that serum NT-proCNP level is likely secondary to existing inflammation. However, the magnitude of CNP action in SSc and its possible role in the pathogenesis of the disease remains to be elucidated. PMID- 20001846 TI - Multiple changes induced by fibroblasts on breast cancer cells. AB - It is now widely recognized that the cross-talk between cancer and stromal cells may play a crucial role in cancer progression. However, little is known about the complex underlying molecular mechanisms that occur within the tumor microenvironment. Fibroblasts are the major stromal cells with multiple roles, especially toward both the extracellular matrix and the neighboring cell population, including neoplastic cells. Consequently, proteomic analyses would provide a wider resource for a better understanding of the potential modulating effects exerted by fibroblasts on cancer cells. In this article we describe the effects of fibroblast stimulation on the breast cancer cell line (8701-BC) proteomics, using a trans-well coculture system. Our results clearly indicate that fibroblasts induce considerable proteomic modulations on 8701-BC, mainly in the cytoskeleton proteins and glycolytic enzymes. Additionally, fibroblast conditioned medium increased neoplastic cell proliferation and invasion with a concurrent upregulation of the c-Myc oncogene. Collectively these results suggest that fibroblast stimulation may enhance the malignant potential of breast cancer cells in vitro. PMID- 20001847 TI - Articular cartilage compression: how microstructural response influences pore pressure in relation to matrix health. AB - Our research investigated the influence of degeneration on both the pore-pressure development and microstructural response of cartilage during indentation with a flat-porous-indenter. Experiments were conducted to link the mechanical and structural responses of normal and degenerate articular cartilage. We found that from the instant of loading the degenerate matrix generated a higher peak hydrostatic excess pore pressure in a shorter period of time than the normal matrix. Following the attainment of this peak value the pore pressure in both tissue groups then gradually decayed toward zero over time, thus demonstrating a classical consolidation response. The microstructural analysis provided a unique insight into the influence of degeneration on the mechanisms of internal stress sharing within the loaded matrix. Both disruption of the articular surface and general matrix destructuring results in an altered deformation field in both the directly loaded and nondirectly loaded regions. It is argued that the higher levels of matrix shear combined with less of the applied load being redirected into the wider cartilage continuum accounts for the elevated levels of peak hydrostatic pore pressure generated in the degenerate matrix. PMID- 20001848 TI - Effect of decorin and dermatan sulfate on the mechanical properties of a neocartilage. AB - Decorin is known to influence the size of collagen fibrils in ligaments and tendons and it has been hypothesized to provide a structural link between collagen fibrils in connective tissues, including cartilage. Coincidently, mechanical properties of skin, ligament, and tendons are altered in decorin knockout mice, suggesting it may influence the structural properties of tissue or tissue matrix organization. To further examine the role of decorin in the extracellular matrix development and subsequent material properties of cartilage, tissue (neocartilage) was grown in a 3D culture model using a pure population of genetically modified chondrocytes stably overexpressing decorin (DCN) or decorin lacking dermatan sulfate (MDCN). An empty vector (CON) served as a virus control. Following generation of the cartilage-like tissues, mechanical properties in tension and compression, collagen fibril diameter, matrix organization, and biochemistry of the tissue were determined. There were no differences between CON and DCN tissues in any parameter measured. In contrast, tissue generated in MDCN cultures was thinner, had higher collagen density, and higher elastic moduli as compared to both CON and DCN tissues. Considering there was no difference in stiffness between CON and DCN tissues, the notion that decorin contributes to the mechanical properties via load transfer was refuted in this model. However, contrasts in the mechanical properties of the MDCN tissue suggest that the dermatan sulfate chains on decorin influences the organization/maturation and resultant mechanical properties of the matrix by as an yet-unidentified regulatory mechanism. PMID- 20001849 TI - Prostate cancer screening: clinical impact of WHO calibration of Beckman Coulter Access prostate-specific antigen assays. AB - BACKGROUND: The goal of this study was to evaluate the clinical impact of using the same total prostate-specific antigen (tPSA) and free PSA (fPSA) assays calibrated with World Health Organization (WHO) materials or with Hybritech Tandem-R calibrator. METHODS: From the initial correlation study that included 150 patients, the clinical impact of the WHO calibration was simulated using a large cohort (n=4548) of referred patients. Interim reports of the European Study of Screening for Prostate Cancer (ERSPC) were used to evaluate the clinical outcomes of patients and the risk of prostate cancer (PCa). RESULTS: WHO calibration of tPSA assays leads to a reduction of about 20% in measured results (tPSA WHO=0.81 tPSA Hybritech+0.04; fPSA WHO=0.78 fPSA Hybritech+0.00; %fPSA WHO=0.92 %fPSA Hybritech+0.00). The simulation showed that the WHO calibration is associated with a risk of missing 15% of PCa. CONCLUSIONS: The discrepancies between the two calibrations lead to significant clinical misinterpretation with decreased detection of PCa if tPSA cut-off thresholds are not adjusted. PMID- 20001850 TI - Improving laboratory efficiencies through significant time reduction in the preanalytical phase. PMID- 20001851 TI - O-beta-N-acetyl-D-glucosaminidase in erythrocytes of Italian air force acrobatic pilots. AB - BACKGROUND: Italian air force acrobatic pilots are occupationally susceptible to oxidative stress damage that can lead to overt signs and symptoms of hypoxia. We propose erythrocyte glycohydrolases as new, sensitive markers to assess oxidative stress. METHODS: We measured erythrocyte concentrations of beta-D-glucuronidase (GCR), hexosaminidase, O-beta-N-acetyl-D-glucosaminidase (O-GlcNAcase), plasma membrane fluidity and plasma hydroperoxides from 19 pilots and compared these to 40 matched healthy subjects. RESULTS: Plasma hydroperoxide concentrations and the erythrocyte ghosts' fluorescence anisotropy were significantly lower in the pilots. Concentrations of GCR, O-GlcNAcase and hexosaminidase in pilots were significantly different from controls, being lower, higher and higher, respectively. CONCLUSIONS: Pilots, in spite of their oxidative stress, are better protected than controls, probably as a result of their physical training and proper diet. Our results confirm that erythrocytes, with their 120-day life span, are a useful model for investigating physiopathological conditions, and glycohydrolases are good markers for monitoring oxidative stress, even in healthy people. PMID- 20001852 TI - Detection and quantitation of Epstein-Barr virus (EBV) DNA in EDTA whole blood samples using automated sample preparation and real time PCR. AB - BACKGROUND: Detection and quantitation of Epstein-Barr virus (EBV) DNA in EDTA whole blood samples has gained significance in the routine diagnostic laboratory. METHODS: In this study, the analytical and clinical performance of the artus EBV RG PCR kit in conjunction with automated sample preparation on the QIAsymphony SP instrument was evaluated. RESULTS: When the accuracy of the new test system was tested, all results were found to be within +/-0.5 log(10) unit of the expected panel results. Determination of linearity showed a quasilinear curve over 4 log units. The lower limit of detection was determined to be 391 EBV DNA copies/mL in EDTA whole blood. The between day imprecision ranged from 18% to 66%, and the within run imprecision ranged from 11% to 50%. When clinical samples were tested and the results compared with those obtained with the routinely used easyMAG sample preparation and EBV R-gene test system, 60 samples tested positive and 31 samples tested negative by both assays. Nineteen samples were found to be positive using the QIAsymphony sample preparation and artus EBV RG PCR test system only, and no samples tested positive with the routinely used test system only. CONCLUSIONS: The QIAsymphony sample preparation and artus EBV RG PCR test system is suitable for the detection and quantitation of EBV DNA in EDTA whole blood in the routine diagnostic laboratory. PMID- 20001853 TI - Factors influencing time to vancomycin-induced clearance of nonendocarditis methicillin-resistant Staphylococcus aureus bacteremia: role of platelet microbicidal protein killing and agr genotypes. AB - BACKGROUND: Vancomycin susceptibility, the accessory gene global regulator (agr) genotype and function, staphylococcal cassette chromosome (SCC) mec type, and susceptibility to cationic thrombin-induced platelet microbicidal protein 1 (tPMP 1) have been individually predictive of duration of methicillin-resistant Staphylococcus aureus (MRSA) bacteremia. This investigation evaluated the interrelationship of these factors with time to clearance of MRSA bacteremia during vancomycin therapy in patients without endocarditis. METHODS: Vancomycin minimum inhibitory concentration and in vitro killing, agr function (delta hemolysin activity), agr group, SCCmec type, and survival in tPMP-1 killing assays were determined for 29 MRSA bacteremia isolates. RESULTS: Increased resistance to tPMP-1 killing was observed with agr group III MRSA (P = .025) and MRSA with reduced or absent agr function (P = .023). The median time to clearance of MRSA bacteremia was earlier for agr group III (3 days) versus group I (10.5 days) or II (15 days) (P = .001). In multivariate analysis, agr group II, reduced tPMP-1 killing in vitro, and prior vancomycin exposure were significant independent predictors of longer MRSA bacteremia duration. CONCLUSIONS: Specific genotypic, phenotypic, and clinical parameters appear to correlate with persistent MRSA bacteremia. The interrelationship of these and other factors probably contributes to vancomycin-mediated clearance of MRSA bacteremia. PMID- 20001854 TI - Immunologic and virologic events in early HIV infection predict subsequent rate of progression. AB - BACKGROUND: Variability in human immunodeficiency virus (HIV) disease progression cannot be fully predicted by CD4(+) T cell counts or viral load (VL). Because central memory T (T(CM)) cells play a critical role in the pathogenesis of simian immunodeficiency virus disease, we hypothesized that quantifying these cells in early HIV infection could provide prognostic information. METHODS: We measured expression of CD45RO, chemokine (C-C motif) receptor (CCR) 5, CCR7, CD27, and CD28 to enumerate naive and memory subsets in samples from recently infected individuals. We also quantified proliferation, CD127 expression, and cell associated VL. Disease progression was compared across subgroups defined by these measurements, using Kaplan-Meier survival curves and multivariate Cox proportional hazards regression. RESULTS: Four hundred sixty-six subjects contributed 101 events. The proportion or absolute count of T(CM) cells did not correlate with disease progression, defined as the time to AIDS or death. However, significant associations were observed for proliferation within CD4(+) or CD8(+) T cells, loss of naive or CD127(+) memory CD8(+) T cells, and CD4(+) T cell-associated VL. CONCLUSIONS: Our results demonstrate that the extent of the immunopathogenesis established early in HIV infection predicts the course of future disease. Because antiretroviral drug treatment reverses such defects in part, our study provides mechanistic clues to why early use of antiretrovirals may prove beneficial. PMID- 20001855 TI - Preservation of memory CD4(+) T lymphocytes in breast milk of lactating rhesus monkeys during acute simian immunodeficiency virus infection. AB - Acute human immunodeficiency virus (HIV) or simian immunodeficiency virus (SIV) infection is associated with a massive depletion of memory CD4(+) T lymphocytes in the gastrointestinal tract. To define the dynamics of the CD4(+) T lymphocyte subpopulations in breast milk during acute HIV or SIV infection, lymphocyte populations were monitored in blood and milk of 4 Mamu-A*01(+) rhesus monkeys after SIVmac251 inoculation. Strikingly, although the CD4(+) T lymphocytes in blood were depleted during the peak of viremia, the milk CD4(+) T lymphocyte counts remained unchanged, despite active virus replication in the breast milk compartment. Moreover, CD4(+) memory T lymphocytes were preserved in breast milk during acute infection. CD4(+) T lymphocytes in breast milk and other mucosal compartments of uninfected monkeys were similar in their memory phenotype, activation status, and chemokine (C-C motif) receptor 5 expression. Interestingly, the number and proportion of effector CD8(+) T lymphocytes in milk were increased during acute SIV infection, suggesting effective control of virus mediated CD4(+) T lymphocyte destruction in the breast milk compartment. PMID- 20001856 TI - No evidence for decay of the latent reservoir in HIV-1-infected patients receiving intensive enfuvirtide-containing antiretroviral therapy. AB - Human immunodeficiency virus type 1 (HIV-1) persists in a latent reservoir of infected resting memory CD4 cells in patients receiving antiretroviral therapy. We assessed whether multitarget therapy with enfuvirtide, 2 reverse-transcriptase inhibitors, and a ritonavir-boosted protease inhibitor leads to decay of this reservoir. Nineteen treatment-naive patients initiated this regimen; 9 experienced virologic suppression and continued enfuvirtide-containing therapy for at least 48 weeks. In enfuvirtide-treated patients with virological suppression, there was no decay of the latent reservoir (95% confidence interval for half-life, 11 months to infinity). The stability of the latent reservoir despite intensive therapy suggests that new strategies are needed to eradicate HIV-1 from this reservoir. (ClinicalTrials.gov identifier: NCT00051831 .). PMID- 20001857 TI - The interleukin 6 -174 C/C genotype predicts greater rhinovirus illness. AB - BACKGROUND: In adults and children with respiratory syncytial virus (RSV) infection, a polymorphism in the interleukin 6 (IL-6) promoter at position -174 predicts illness magnitude. In addition, polymorphisms in the interleukin 10 (IL 10), tumor necrosis factor alpha (TNF-alpha), and interferon gamma (IFN-gamma) genes are associated with immune responsiveness and the frequency of complications. Here, the effect of these polymorphisms on illness and seroconversion during infection with rhinovirus type 39 (RV39) was evaluated. METHODS: Seventy-two adults were genotyped for the selected polymorphisms, experimentally exposed to RV39, and followed to track infection, seroconversion, and symptoms and signs of illness. Regression analysis was used to determine whether these polymorphisms predicted seroconversion and illness magnitude in 57 infected subjects. RESULTS: The low-production IL-6 -174 phenotype (C/C genotype) was associated with greater symptom magnitudes, and the IFN-gamma phenotype +874 predicted the frequency of seroconversion. No relationship between the IL-10 or TNF-alpha polymorphisms and any measured outcome was documented. The concentration of IL-6 protein, as measured in nasal wash fluids from subjects, was positively correlated with symptom magnitude, but it was independent of the IL-6 -174 genotypes representing the high- and low-production phenotypes. CONCLUSIONS: These results document statistically significant associations between the IL-6 -174 and IFN-gamma +874 polymorphisms and specific responses to experimental RV39 infection. For the IL-6 -174 polymorphism, the results replicate those for experimental RSV infection. PMID- 20001859 TI - OMICS: 2009, 2010, and Beyond. AB - A Journal of Integrative Biology has been growing by leaps and bounds, and I would like to share some of this with you, our authors and readers. PMID- 20001858 TI - SaeR binds a consensus sequence within virulence gene promoters to advance USA300 pathogenesis. AB - This investigation examines the role of the SaeR/S 2-component system in USA300, a prominent circulating clone of community-associated methicillin-resistant Staphylococcus aureus. Using a saeR/S isogenic deletion mutant of USA300 (USA300DeltasaeR/S) in murine models of sepsis and soft-tissue infection revealed that this sensory system is critical to pathogenesis of USA300 during both superficial and invasive infection. Oligonucleotide microarray and real-time reverse-transcriptase polymerase chain reaction identified numerous extracellular virulence genes that are down-regulated in USA300DeltasaeR/S. Unexpectedly, an up regulation of mecA and mecR1 corresponded to increased methicillin resistance in USA300DeltasaeR/S. 5'-RACE analysis defined transcript start sites for sbi, efb, mecA, lukS-PV, hlb, SAUSA300_1975, and hla, to underscore a conserved consensus sequence within promoter regions of genes under strong SaeR/S transcriptional regulation. Electrophoretic mobility shift assay experiments illustrated direct binding of SaeR(His) to promoter regions containing the conserved consensus sequence. Collectively, the findings of this investigation demonstrate that SaeR/S directly interacts with virulence gene promoters to significantly influence USA300 pathogenesis. PMID- 20001860 TI - Proteomic analysis of bone tissues of patients with osteonecrosis of the femoral head. AB - Osteonecrosis of the femoral head (ONFH) is a devastating disease that can result in a femoral head collapse. By proteomics analysis, we identified 1,967 proteins with two or more unique peptides from ONFH and from control bones with a false discovery rate of 4.8%. Using spectral counting, we identified 141 overexpressed and 56 underexpressed proteins comparing ONFH bones to the controls. GSEA (gene set enrichment analysis) revealed that proteins overexpressed in ONFH are enriched for gene sets related to multiple myeloma and adult T-cell lymphoma (ATL), and to JAK2-dependent genes. We confirmed the underexpression of CHST2 (isoform 1 of carbohydrate sulfotransferase 2), a key protein involved in biosynthesis of chondroitin sulfate proteoglycans, and the underexpression of GPCR26 (G-protein coupled receptor 26), a protein that mediates intracellular calcium mobilization, in ONFH bones compared to controls. Taken together, our data suggest that biosynthesis of chondroitin sulfate proteoglycans and cation transport and mobilization may be a key process involved in the pathogenesis of ONFH. Our analysis sheds new light on the understanding of the pathogenesis of ONFH. PMID- 20001861 TI - Involvement of annexin A1 in multidrug resistance of K562/ADR cells identified by the proteomic study. AB - Multidrug resistance (MDR) to chemotherapy is a significant barrier to the effective treatment of chromic myeloid leukemia (CML). In an attempt to identify more factors associated with MDR for an understanding of the mechanism, we first established an adriamycin (ADR)-resistant human erythroleukemia cell line K562/ADR by stepwise selection in vitro using ADR. Besides the elevated resistance to ADR, the K562/ADR cells also showed significantly increased crossed resistance to vincristin and Gleevec, compared to the parental K562 cells. Then we compared the global protein profiles between K562 and K562/ADR cells. Following two-dimensional gel electrophoresis and image analysis, some of the proteins with different levels between the two cell lines were identified by MALDI TOF/TOF mass spectrometry and Western blot analysis. The differentially expressed proteins were classified into groups based on their functions: calcium binding proteins, chaperones, metabolic enzymes, proteins related to protein synthesis or DNA synthesis, and proteins related to signal transduction. In particular, ANXA1, a protein that was downregulated in K562/ADR, was analyzed further for its involvement in MDR by transfection and subsequent assays. The functional validation showed that the downregulated ANXA1 expression contributes considerably to the observed drug resistance in K562/ADR cells. These data will be valuable for further study of the mechanisms of MDR and may reveal a potential new diagnostic marker to chemotherapy. PMID- 20001862 TI - GeneDecks: paralog hunting and gene-set distillation with GeneCards annotation. AB - Sophisticated genomic navigation strongly benefits from a capacity to establish a similarity metric among genes. GeneDecks is a novel analysis tool that provides such a metric by highlighting shared descriptors between pairs of genes, based on the rich annotation within the GeneCards compendium of human genes. The current implementation addresses information about pathways, protein domains, Gene Ontology (GO) terms, mouse phenotypes, mRNA expression patterns, disorders, drug relationships, and sequence-based paralogy. GeneDecks has two modes: (1) Paralog Hunter, which seeks functional paralogs based on combinatorial similarity of attributes; and (2) Set Distiller, which ranks descriptors by their degree of sharing within a given gene set. GeneDecks enables the elucidation of unsuspected putative functional paralogs, and a refined scrutiny of various gene-sets (e.g., from high-throughput experiments) for discovering relevant biological patterns. PMID- 20001863 TI - CHF1/Hey2 promotes physiological hypertrophy in response to pressure overload through selective repression and activation of specific transcriptional pathways. AB - We have previously found that CHF1/Hey2 prevents the development of phenylephrine induced cardiac hypertrophy. To determine the role of CHF1/Hey2 in pressure overload hypertrophy, we performed ascending aortic banding on wild-type and transgenic mice overexpressing CHF1/Hey2 in the myocardium. We found that both wild-type and transgenic mice developed increased ventricular weight to body weight ratios 1 week after aortic banding. Wild-type mice also developed decreased fractional shortening after 1 week when compared to preoperative echocardiograms and sham-operated controls. Transgenic mice, in comparison, demonstrated preserved fractional shortening. Histological examination of explanted heart tissue demonstrated extensive fibrosis in wild-type hearts, but minimal fibrosis in transgenic hearts. TUNEL staining demonstrated increased apoptosis in the wild-type hearts but not in the transgenic hearts. Exposure of cultured neonatal myocytes from wild-type and transgenic animals to hydrogen peroxide, a potent inducer of apoptosis, demonstrated increased apoptosis in the wild-type cells. Gene Set Analysis of microarray data from wild-type and transgenic hearts 1 week after banding revealed suppression and activation of multiple pathways involving apoptosis, cell signaling, and biosynthesis. These findings demonstrate that CHF1/Hey2 promotes physiological over pathological hypertrophy through suppression of apoptosis and regulation of multiple transcriptional pathways. These findings also suggest that CHF1/Hey2 and its downstream pathways provide a variety of targets for novel heart failure drug discovery, and that genetic polymorphisms in CHF1/Hey2 may affect susceptibility to hypertrophy and heart failure. PMID- 20001865 TI - Sorafenib - a small molecule with big promise? PMID- 20001866 TI - New agents for the treatment of genetically high-risk chronic lymphocytic leukemia. PMID- 20001867 TI - Bevacizumab for advanced breast cancer. AB - Tumor angiogenesis is an important step in breast cancer development, progression, invasion and metastasis. Pro-angiogenic factors such as VEGF regulate angiogenesis and are targets for drug development. Bevacizumab, an anti VEGF antibody, has demonstrated significant clinical benefit in several solid tumors, including breast cancer. Its use in combination with either paclitaxel or docetaxel has prolonged progression-free survival and increased response rates in the first-line treatment of patients with metastatic breast cancer. This review article discusses the clinical trials establishing the use of bevacizumab for the treatment of advanced breast cancer. PMID- 20001868 TI - Dienogest: a new therapeutic agent for the treatment of endometriosis. AB - Dienogest (DNG), a progestin of 19-nortestosterone derivative, has good oral bioavailability and is highly selective for progesterone receptors. Owing to its antiovulatory, antiproliferative activities in endometrial cells, and its inhibitory effects on the secretion of cytokines, DNG is expected to be an effective treatment for endometriosis. Progesterone receptor-binding affinity is higher for DNG than for progesterone. Several pilot studies demonstrated that after 24 weeks of DNG treatment, there was a significant decrease in terms of dysmenorrhea, premenstrual pain, dyspareunia and diffuse pelvic pain. Most of the cases of genital bleeding occurring in the DNG treatment were spotting or breakthrough bleeding, which decreased with continued treatment and resolved either during treatment or after the end of treatment. The therapeutic effects of DNG 2 mg/day and norethisterone acetate 10 mg/day for endometriotic symptoms during a period of 24 weeks were almost similar. The only disadvantage of DNG seems to be the irregular bleeding. Good efficacy and tolerability of DNG in patients with endometriosis have been demonstrated in an open, randomized European clinical trial as compared with norethisterone acetate. In Japan, a Phase III, randomized, double-blind, multicenter, controlled trial was conducted to compare the efficacy and safety of DNG with intranasal buserelin acetate in patients with endometriosis. The study demonstrated that DNG is as effective as intranasal buserelin acetate in alleviating endometriosis symptoms, and causes less bone mineral density loss, resulting in the use on a commercial basis for endometriosis patients in Japan from 2008. This paper provides summarized data on this new promising drug for endometriosis. PMID- 20001869 TI - Should we question if something works just because we don't know how it works? PMID- 20001870 TI - Oxygen delivery to patients after cardiac surgery: a medical record audit. AB - OBJECTIVE: To describe how intensive care nurses manage the administration of supplemental oxygen to patients during the first 24 hours after cardiac surgery. METHODS: A retrospective audit was conducted of the medical records of 245 adult patients who underwent cardiac surgery between 1 January 2005 and 31 May 2008 in an Australian metropolitan hospital. Physiological data (oxygen saturation measured by pulse oximetry and respiratory rate) and intensive care unit management data (oxygen delivery device, oxygen flow rate and duration of mechanical ventilation) were collected at hourly intervals over the first 24 hours of ICU care. RESULTS: Of the 245 patients whose records were audited, 185 were male; mean age was 70 years (SD, 10), and mean APACHE II score was 17.5 (SD, 5.14). Almost half the patients (122, 49.8%) were extubated within 8 hours of ICU admission. The most common oxygen delivery device used immediately after extubation was the simple face mask (214 patients, 87%). Following extubation, patients received supplemental oxygen via, on average, two different delivery devices (range, 1-3), and had the delivery device changed an average of 1.38 times (range, 0-6) during the 24 hours studied. Twenty-two patients (9%) received non-invasive ventilation or high-flow oxygen therapy, and 16 (7%) experienced one or more episode of hypoxaemia during mechanical ventilation. A total of 148 patients (60%) experienced one or more episodes of low oxygenation or abnormal respiratory rate during the first 24 hours of ICU care despite receiving supplemental oxygen. CONCLUSION: These findings suggest that the ICU environment does not protect cardiac surgical patients from suboptimal oxygen delivery, and highlights the need for strategies to prompt the early initiation of interventions aimed at optimising blood oxygen levels in cardiac surgical patients in the ICU. PMID- 20001871 TI - Prospective observational study of postoperative complications after percutaneous dilatational or surgical tracheostomy in critically ill patients. AB - OBJECTIVE: To assess and describe postoperative complications of single dilator percutaneous dilatational tracheostomy (PDT) and surgical tracheostomy (ST) in a large series of critically ill patients. METHODS: A prospective observational study was conducted in 1163 critically ill patients in a university affiliated tertiary referral hospital between 2002 and 2007. PDT was the procedure of choice for all critically ill patients requiring tracheostomy except for those with an anatomic abnormality or refractory coagulopathy, who underwent ST. Demographic and postoperative complication data were collected in a web-based database. RESULTS: 913 patients (79%) underwent PDT at the bedside in the ICU, and 250 (21%) underwent ST in the operating theatre. The tracheostomy tube was larger, and the duration of tracheostomy cannulation was shorter after PDT than after ST. The postoperative complication rate for PDT was 9.6% compared with 19.6% for ST (P<0.001). Tracheal tube obstruction and displacement were significantly less frequent after PDT (obstruction 1.0% for PDT v 3.6% for ST, P = 0.007; displacement, 1.3% for PDT v 4.8% for ST, P = 0.002). CONCLUSIONS: In a large heterogeneous group of critically ill patients, single dilator PDT was safe and had few postoperative complications. Although ST was used in higher-risk patients, those who underwent PDT were more likely to receive a larger-sized tracheostomy tube; they were also less likely to experience obstruction or displacement of the postoperative tracheostomy tube. These differences are probably related to a combination of patient selection, smaller, shorter tracheostomy tubes, and larger tissue incision size with ST. PMID- 20001873 TI - Predicting future intensive care demand in Australia. AB - BACKGROUND: Predicting future demand for intensive care is vital to planning the allocation of resources. METHOD: Mathematical modelling using the autoregressive integrated moving average (ARIMA) was applied to intensive care data from the Australian and New Zealand Intensive Care Society (ANZICS) Core Database and population projections from the Australian Bureau of Statistics to forecast future demand in Australian intensive care. RESULTS: The model forecasts an increase in ICU demand of over 50% by 2020, with current total ICU bed-days (in 2007) of 471 358, predicted to increase to 643 160 by 2020. An increased rate of ICU use by patients older than 80 years was also noted, with the average bed-days per 10 000 population for this group increasing from 396 in 2006 to 741 in 2007. CONCLUSION: An increase in demand of the forecast magnitude could not be accommodated within current ICU capacity. Significant action will be required. PMID- 20001874 TI - Automated external defibrillators and survival after in-hospital cardiac arrest: early experience at an Australian teaching hospital. AB - OBJECTIVE: To evaluate the effect of the introduction of automated external defibrillators (AEDs) on survival after inhospital cardiac arrest. DESIGN, SETTING AND PARTICIPANTS: Before-and-after study that compared patients during the 2 years before (8 November 2005 to 7 November 2007) and the year after (8 November 2007 to 7 November 2008) the deployment of AEDs to the non-critical care areas of a university teaching hospital. MAIN OUTCOME MEASURES: Return of spontaneous circulation (ROSC) and survival to hospital discharge. RESULTS: 55 in hospital cardiac arrests occurred in the 2-year pre-AED period and 31 in the 1 year AED period. Patients had similar baseline characteristics in the pre-AED and AED periods including witnessed arrest (53% v 48%), arrest in an acute inpatient ward (78% v 90%), and initial arrest rhythm of pulseless ventricular tachycardia or ventricular fibrillation (18% v 16%). The proportions of patients with ROSC were similar in the pre-AED and AED periods (42% v 55%), as were the proportions who survived to hospital discharge (22% v 29%). In the AED period, the relative risk of ROSC was 1.31 (95% CI, 0.84- 2.04) and the relative risk of survival to hospital discharge was 1.33 (95% CI, 0.63-2.80). CONCLUSIONS: ROSC and survival to hospital discharge did not change significantly after deployment of AEDs. The existence of a timely and robust resuscitation response with relatively good baseline outcomes, and the low proportion of initial shockable arrest rhythms may have limited the capacity of AEDs to improve survival. PMID- 20001875 TI - Withholding and withdrawal of life-sustaining therapies in intensive care: an Australian experience. AB - OBJECTIVES: Withholding and withdrawal of treatment in intensive care is currently widely accepted, but little has been published about Australian practice. DESIGN AND SETTING: Retrospective audit of all deaths in two major tertiary intensive care units in the Hunter New England Area Health Service during 2008. Patients who died were classified as "no limitations" (died while receiving full treatment), "treatments withheld" (specific treatment limitations) or "withdrawal of life-sustaining treatment" (WLST). RESULTS: Of 1950 patients admitted to an ICU, 283 died (14.5%). Of these 283, 54 (19%) died despite all therapeutic efforts; 97 (34%) had treatments withheld, and 132 (47%) had WLST. There were no statistically significant differences in length of stay between the three groups. Patients who died despite all therapeutic efforts were younger than those who died after treatments were withheld or WLST (mean age [SD], 58.7 [21.1] v 73.1 [12.5] v 69.3 [13.5]; P = 0.001). APACHE II score was higher in the group who died than in the total ICU group (mean [SD], 25.5 [8.3] v 17.7 [8.7], P = 0.001). CONCLUSIONS: In this population of critically ill patients, most deaths occurred after discussion of end-of-life decisions and withholding or withdrawal of treatment. PMID- 20001872 TI - Effects of saline or albumin resuscitation on standard coagulation tests. AB - AIMS: To explore whether fluid resuscitation with normal saline or 4% albumin is associated with differential changes in routine clinical coagulation tests. DESIGN: Substudy from a large double-blind randomised controlled trial, the SAFE (Saline versus Albumin Fluid Evaluation) study. SETTING: Three general intensive care units. PATIENTS: Cohort of 687 critically ill patients. INTERVENTION: We randomly allocated patients to receive either 4% human albumin or normal saline for fluid resuscitation, and collected demographic and haematological data. METHODS AND MAIN RESULTS: Albumin was administered to 338 patients and saline to 349. At baseline, the two groups had similar mean activated partial thromboplastin time (APTT) of 37.2 s (albumin) v 39.1 s (saline); mean international normalised ratio (INR) of 1.38 v 1.34, and mean platelet count of 244 x 10(9)/L v 249 x 10(9)/L. After randomisation, during the first day of treatment, the APTT in the albumin group was prolonged by a mean of 2.7 s, but shortened slightly by a mean of -0.9 s in the saline group. The INR did not change in either group, while the platelet count decreased transiently in both groups. Using multivariate analysis of covariance to account for baseline coagulation status, albumin fluid resuscitation (P = 0.01) and a greater overall volume of resuscitation (P = 0.03) were independently associated with prolongation of APTT during the first day. CONCLUSIONS: Administration of albumin or of larger fluid volumes is associated with a prolongation of APTT. In ICU patients, the choice and amount of resuscitation fluid may affect a routinely used coagulation test. PMID- 20001876 TI - Emphysematous pyelonephritis: an unusual complication of blunt abdominal trauma. AB - We report an unusual case of emphysematous pyelonephritis after blunt abdominal trauma in a 28-year-old woman. She was previously healthy and did not have diabetes. Treatment with antibiotics and percutaneous drainage was successful. We review the diagnostic and management strategies for this condition. PMID- 20001877 TI - Prolonged serotonin toxicity with proserotonergic drugs in the intensive care unit. AB - Serotonin toxicity secondary to drug therapy, interaction or overdose is an increasing phenomenon worldwide. A proportion of patients require admission to an intensive care unit, but the treatment needed is usually supportive and of short duration. Prolonged ICU admission to control ongoing or long-lasting serotonin toxicity has not been reported previously. We describe three patients with prolonged serotonin toxicity, lasting 12-18 days. Symptoms of toxicity were easily demonstrable in each and were refractory to currently recommended therapies. We review the pharmacological mechanisms that led to prolonged serotonin toxicity in these patients. Predictors for prolonged serotonin toxicity include involvement of irreversible monoamine oxidase inhibitors (MAOIs) or slow release preparations resistant to the effects of activated charcoal (eg, lithium). We also discuss the implications of prolonged toxicity for critical care management, to maintain optimal patient outcomes. PMID- 20001878 TI - A new paradigm for treating infections: "go hard and go home". AB - There is now significant evidence that initial use of the correct antibiotic saves more lives than virtually all other intensive care therapy. This means covering all possible causative organisms with the initial empirical choice. For nosocomial sepsis, broad-spectrum antibiotics must be started as soon as the relevant samples have been taken for culture, with de-escalation of therapy targeted to the causative organisms when results and susceptibilities are available. There is an international trend to use shorter antibiotic courses. Pseudomonas pneumonia probably needs a 7-10 day course. In our ICU, provided the infection source is controlled, we seldom use antibiotic courses longer than 7 days. Evaluation of the kill characteristics of antibiotics in experimental models suggests that different classes of antibiotics should have different dosing regimens. For Beta- lactam antibiotics, the kill characteristic is almost entirely related to the time that tissue and plasma levels exceed a certain threshold, with no significant post-antibiotic effect, particularly against gram negative organisms. Kill characteristics of other antibiotics, such as aminoglycosides, relate to adequate peak concentrations and a significant post antibiotic effect. Clinically, these kill characteristics translate into the need for appropriate doses of the various antibiotics in patients with sepsis. We have shown that some patients with normal serum creatinine levels have very high creatinine clearance rates; in ICU patients with sepsis, blood pressure and tissue perfusion are maintained with large fluid loads and inotropic agents, thereby raising creatinine clearance. High clearances produce low trough concentrations of antibiotic, with important implications for underdosing and the development of antibiotic resistance. The new paradigm for treating sepsis, particularly nosocomial sepsis, is: get it right the first time, hit hard up front, and use large doses of broad-spectrum antibiotics for a short period. PMID- 20001879 TI - New antimicrobial agents for methicillin-resistant Staphylococcus aureus. AB - In bacterial and fungal infections, optimal outcomes are obtained through the timely provision of adequate antimicrobial coverage in an initial anti-infective treatment regimen. However, selecting appropriate antimicrobial regimens to treat infections in the intensive care unit is challenging because of the expansion of antibiotic resistance. Multidrug anti-infective regimens are typically needed to adequately cover common important pathogens in ICUs. Here, we describe novel antibacterial agents in the late stages of clinical development that show potential for treating methicillin-resistant Staphylococcus aureus (MRSA) infections. These include the fifth-generation cephalosporins, ceftaroline and ceftobiprole; the glycopeptides, dalbavancin, oritavancin, and telavancin; and iclaprim. PMID- 20001880 TI - Assessment of tissue cortisol activity. AB - The concept of relative adrenal insufficiency in patients with severe sepsis continues to be controversial. This arises in part from the lack of an accepted "gold standard" for the diagnosis of adrenal insufficiency in the critically ill. Historically, assessment of adrenal function in this population has relied on measurement of plasma total cortisol level, in a blood sample taken either at random or as part of a corticotropin stimulation test. However, an alternative is to focus on the site of glucocorticoid activity within the tissues as a potentially more useful index of functional adrenal status. We review the mechanisms known to affect tissue glucocorticoid activity and examine how they may be modified by critical illness. These include both free and interstitial cortisol concentrations, intracellular cortisol generation, and glucocorticoid receptor activity and density. Changes in these factors are not reflected in plasma total cortisol concentrations, and more sophisticated techniques, including genetic transcriptional surveys, may be required to reveal the role of glucocorticoid insufficiency in critical illness. PMID- 20001881 TI - Should we mobilise critically ill patients? A review. AB - BACKGROUND: Neuromuscular weakness, a frequent complication of prolonged bed rest and critical illness, is associated with morbidity and mortality. Mobilisation physiotherapy has widespread application in patients hospitalised with non critical illness. OBJECTIVES: We reviewed the literature to evaluate the worldwide availability of mobilisation therapy in intensive care units and the role of mobilisation therapy in patients requiring medical or surgical high dependency or intensive care. METHODS: We searched PubMed (1980 to August 2009) using the MeSH terms "physiotherapy" and "intensive care". Additional keyword search terms, "mobilisation", "mobilization", and "fast-track", were used. In addition, we examined reference lists in recent studies and reviews. RESULTS: Routine mobilisation physiotherapy is least likely to be available in ICUs in the United States. Early mobilisation is appropriate for patients with pulmonary thromboembolic disease, community-acquired pneumonia and in elderly hospitalised patients. Although fast-track cardiac and noncardiac surgery with early ambulation is safe and reduces hospital length of stay, it does not alter postoperative mortality. Up to 25% of patients can be safely mobilised within 72 hours of ICU admission. This therapy may reduce hospital and ICU length of stay, shorten duration of mechanical ventilation, and improve muscle strength and functional independence scores. Pooled data show a nonsignificant mortality benefit in favour of early mobilisation (odds ratio, 0.77; 95% CI, 0.49-1.21). CONCLUSIONS: The data in support of mobilisation therapy for perioperative and critically ill patients, while of a low level of evidence, are substantial. This justifies a paradigm shift in attitudes towards physiotherapy and the prevention of critical illness weakness. PMID- 20001882 TI - Sick adrenal or sick euadrenal? AB - The role of glucocorticoid supplementation in septic shock remains contentious. In septic shock, the driver for steroid therapy is the premise that there is relative adrenal insufficiency (based on reduced plasma cortisol and blunted cortisol response to corticotropin). The uncertainty arises from the inability of current tests to clearly identify patients who are truly corticosteroid "deficient" at a cellular level, and hence require supplemental glucocorticoid administration. We hypothesise that plasma measurements (total plasma cortisol level and the response to corticotropin) do not consistently reflect the functional adrenal response to stress. Published evidence indicates that there are cellular adaptations in stress, such as pre-receptor upregulation of cortisol, altered receptor density and gene transcription changes, none of which are reflected by plasma cortisol level. This leads us to postulate that the lack of a clearly defined plasma response in severe stress and the presence of an adequate response at the cellular level suggest it is a "sick euadrenal state", analogous to the sick euthyroid state, and not a sick adrenal indicating adrenal insufficiency. PMID- 20001883 TI - Forget glucose: what about lipids in critical illness? AB - A high serum cholesterol level is a risk factor for cardiovascular disease and has commonly been linked with worse outcomes. It is now well recognised that, in many critically ill patients, the opposite is true, with hypocholesterolaemia being associated with poor outcomes. In critical illness, particularly sepsis, total and high-density lipoprotein (HDL) cholesterol levels are commonly decreased, with varying changes in triglyceride levels. The magnitude of the changes seems to reflect the severity of inflammation. Plausible biological explanations exist to explain these associations, including an interaction of lipoproteins with endotoxin and the regulation of cytokine production. It remains unclear whether these observed alterations in lipid profile are a consequence of the physiological disturbance or whether they have a more causative role, worsening organ dysfunction or predisposing to infection. Lipid emulsions provide a vehicle for drug delivery, have become an important part of nutrition, and are emerging as a therapy for specific intoxications. The nature, dietary source and amount of lipid provided to critically ill patients may be enormously important and warrant more rigorous investigation. Further understanding of the alterations in lipid metabolism may have therapeutic implications in treatment of sepsis with specific compounds that manipulate lipid profiles, such as fibrates, statins, niacin and even reconstituted HDL. PMID- 20001885 TI - The association between preoperative eGFR and outcomes in cardiac surgical patients. PMID- 20001886 TI - Genital and oral mucosal immune response against HIV-1 in exposed uninfected individuals. AB - Epidemiologic reports routinely indicate that the high rate of HIV-1 transmission via mucosal exposure has been relatively stable since the epidemic began. Unfortunately, research on mucosal immune responses to HIV-1 has not been done in proportion to its importance. Most knowledge about immune responses against HIV-1 in humans comes from studies limited to the use of peripheral blood cells and plasma. Consequently, T-cell-based HIV-1 vaccines have long been considered a primary end point of preventive therapeutic strategies. The interest in HIV-1 exposed uninfected individuals has intensified because of the lessons to be learned about a natural immunologic response that promotes opposition to the infection. Such information has useful applications in the clinical setting. This review describes the current status of research on mucosal immune responses to HIV-1 from examining mucosal fluids and tissues of sexually exposed uninfected adults. PMID- 20001887 TI - A chimerism-based approach to induce tolerance in IgE-mediated allergy. AB - Immunoglobulin-E-mediated allergy (type I allergy) is a T-helper-2-mediated disease with increasing prevalence in industrialized countries. Immunotherapy is available as causative treatment, but an effective preventive strategy is still an unmet need. Molecular chimerism is an attractive experimental approach that induces tolerance through transplantation of autologous hematopoietic stem cells that are genetically modified to express the disease-causing antigen(s). Molecular chimerism leads to permanent and robust tolerance in experimental models of autoimmune diseases and organ transplantation. Recently, proof-of principle studies demonstrated that a type I allergic immune response can be durably tolerized by transplantation of allergen-expressing syngeneic bone marrow. We review the concept of tolerance induction through chimerism and discuss the potential of this strategy in immunoglobulin-E-mediated allergy. PMID- 20001888 TI - Current vaccination strategies for the treatment of B-cell lymphoma and multiple myeloma. AB - Therapeutic vaccines have been developed to induce immune responses capable of eradicating lymphoma and myeloma tumors. Most of these vaccines target the immunoglobulin idiotype (Id) as a tumor-specific antigen. Phase I/II clinical trials of Id vaccination in lymphoma demonstrated that some lymphoma patients could mount immune responses that were correlated with a favorable clinical outcome. These encouraging results initiated phase III trials of Id vaccination in lymphoma, the results of which have been recently released. Disappointingly, only one of three phase III studies achieved the primary end point of progression free survival. Detailed analysis of these results is awaited to help identify factors that determine clinical efficacy of Id vaccination as reflected in the three trials. Unlike lymphoma, studies of Id vaccination in multiple myeloma have yielded far less evidence of clinical benefit. Although Id vaccines induce immune responses in myeloma patients, their efficacy is insufficient to provide a clear clinical benefit. Strategies to improve lymphoma and myeloma vaccines are currently tested with an emphasis on optimization of antigen delivery and presentation and modulation of the immune system toward enhancement of T-cell function. Despite many hurdles yet to overcome, it is hoped that newly developed strategies that augment both immune and clinical responses will allow effective vaccination resulting in tumor eradication. PMID- 20001889 TI - Opposing forces in asthma: regulation of signaling pathways by kinases and phosphatases. AB - Allergic asthma is a chronic inflammatory disease that involves a sustained T helper-2 (Th2) type immune response and the recruitment of inflammatory cells such as lymphocytes, mast cells, and eosinophils to the lung. The complex cellular interactions involved in this disease are dependent on cellular responses generated following the integration of environmental signals through the cell signaling pathways. Therefore, the elucidation of the biochemical cascades involved in cellular responses contributing to asthma development has been of great interest in this field. Much attention has been given to the activities of kinases, enzymes that catalyze the addition of a phosphate group to a protein, thus allowing them to modify the function of the target enzyme. This resulted in the identification of key kinases and various cellular processes involved in disease development. New investigations have also begun to unravel the importance of phosphatases, which catalyze the removal of a phosphate group from their target protein. Together, these studies reveal a signaling picture in allergic asthma that is far more complex than originally thought. Herein, we review critical mechanisms of asthma development and discuss how kinases and phosphatases are likely to regulate the development of disease through their effect on these various mechanisms. PMID- 20001890 TI - Paclitaxel in cancer treatment: perspectives and prospects of its delivery challenges. AB - Paclitaxel (PTX) is a potent anticancer agent whose clinical usefulness is marred by a delivery problem that is caused by its unfavorable pharmacokinetic and physical properties. Paclitaxel is currently formulated in a mixture of Cremophor EL and ethanol, which is diluted 5-20 times with normal saline or 5% dextrose prior to administration via slow infusion to avoid precipitation in plasma. Many adverse reactions to the PTX formulation have been reported because of the presence of Cremophor EL, including hypersensitivity reactions, nephrotoxicity, and neurotoxicity. Cremophor EL also causes vasodilation, labored breathing, lethargy, hypotension, and leaching of plasticizers, such as diethylhexylpthalate, from the polyvinylchloride infusion bags/sets. Significant research efforts have been conducted to develop an alternative formulation approach to increase the aqueous solubility of PTX without using Cremophor, thereby decreasing its toxicity. This article reviews the various investigated formulation approaches including pastes; liposomes; conjugates with antibodies, peptides, and fatty acids; nanospheres and microspheres; cyclodextrin complexes; emulsions; mucoadhesive gel; prodrugs; and nanoparticulate systems. The pros and cons of each approach are also discussed. Finally, this review concludes with a discussion of nanoparticulate delivery, which is the most promising PTX delivery system of the future because it incorporates the benefits of other approaches such as conjugation, complexation, and prodrugs. PMID- 20001891 TI - Pharmacoscintigraphy: a blazing trail for the evaluation of new drugs and delivery systems. AB - The drug development process is becoming increasingly sophisticated; as a result, the need for rapid and accurate means of predicting the in vivo behavior of drug products continues to grow. Preclinical in vivo evaluations involve the sacrifice of many animals and hence raise ethical issues. Therefore, noninvasive methods for assessing the in vivo behavior of drug products in animals have become an attractive alternative. Gamma scintigraphy is an established radiological-imaging technique that has been used in medical practice for the past several decades. Gamma scintigraphy is also used during clinical evaluations to map the formulation as it traverses the human body in real time. Combining this information with the pharmacokinetic data gives valuable information about the release and absorption mechanisms of drugs from their products. This technique also provides a handy tool for evaluating formulations at the preclinical and clinical stages of development for researchers who work in targeted drug delivery using novel drug carrier systems. The current review explores the use of pharmacoscintigraphy, a science that integrates gamma scintigraphy and conventional pharmacokinetics in the process of drug development. PMID- 20001892 TI - Specific shortcomings of endograft design. AB - Further to the rapid enlargement of an aneurysm to 5.6 cm in diameter after 3 years of surveillance, a 79-year-old patient was fitted with a Vanguard modular stent graft and monitored on a regular basis for 6 years. Two years later, the aneurysmal sac ruptured. The patient died 1 month after an open surgery. The device was devoid of any encapsulation and the ipsilateral limb was detached from the body. The Nitinol skeleton was mostly maintained, however, some polypropylene sutures were broken. The resulting motion of the sharp-angled Nitinol wires caused abrasion and resulted in a few localized holes that were sufficient to permit blood to percolate through the textile wall. Some polyester yarns in the warp direction were ruptured. The Nitinol wire used in this device was shown to be corrosion resistant but the selection of the polypropylene suture was inappropriate. Because this technology is maturing rapidly, these weaknesses can be avoided in the future generations of endovascular devices. It is recommended that these Nitinol wires be sutured to the fabric and that polyester yarns stronger than 68 decitex in tubes 8 mm in diameter are selected. PMID- 20001894 TI - Implanted electrodes in peripheral nerve stimulation and recording: prospects of their application in electronic prosthesis design. AB - The use of peripheral nervous electrodes over the past 70 years has greatly enhanced understanding of the physiological mechanisms underlying nervous system function. The clinical implementation of these electrodes can restore some of the lost functions of the paralyzed or quadriplegic patient, such as bladder control, arm movements, standing, or walking. Stimulation without appropriate control does not lend itself to clinically useful application. Therefore, peripheral nervous electrodes are also required to collect and decode peripheral nerve impulse recordings. The position, shape, and size of an electrode with respect to a neuron or axon determine the stimulating or recording properties. The use of such coded information from impulse recordings is profoundly important for the control of an electronic prosthesis. This article reviews recently published scientific reports pertaining to the use of peripheral nervous electrodes in order to summarize the classifications found in the literature, delineate the progress of the peripheral nervous microelectrode, and estimate its utilizable prospect in the control of electronic prostheses. PMID- 20001893 TI - Persistent type II endoleak unrelated to an Anaconda aortic stent graft fulfilling the 3Bs requirements of biofunctionality, biodurability, and biocompatibility. AB - A patient was fitted with an Anaconda stent graft for which there was a persistent type II endoleak. Two subsequent attempts at embolization were unable to resolve the endoleak. The diameter of the aneurysm varied initially from 5.5 cm in diameter down to 4.8 cm but then later re-dilated to 6.1 cm, with evidence of persistent flow into the aneurysmal sac from the inferior mesenteric artery. Results from serial computed tomography scans demonstrated clear evidence of a type II endoleak that originated from the inferior mesenteric artery with outflow to a distal lumbar artery. The harvested stent graft did not show evidence of a device-related failure. The stent graft and its modular segments were found to have been properly deployed. Only a thin external capsule was evident at explantation. The internal wall of the device showed irregular and thin encapsulation with scattered mural thrombi, which were more prominent at the bifurcation of the main body of the device. Blood deposits and tissue development were sufficient to prevent blood oozing through the wall. The explanted Anaconda stent graft was devoid of any construction flaws or damage (fatigue of the textile or corrosion of the Nitinol wires) after implantation. PMID- 20001895 TI - Megaprosthetic reconstruction for malignant bone tumors: complications and outcomes. AB - Limb salvage surgery and reconstruction of bone defects is challenging in orthopaedic oncology. Allogenic and autologous bone grafting are well-recognized methods for bone reconstruction. Custom-made and modular, cemented and cementless, mega-prostheses provide the modular options needed intraoperatively to bridge variable bone defects. However, these mega-prostheses have worse long term results compared to conventional total joint replacements. Mechanical failure, aseptic loosening, infection, dislocation, skin necrosis, extensor mechanism ruptures and periprosthetic fractures are acknowledged complications of orthopaedic oncology procedures. These complications are due to the large defects that need to be bridged with a more difficult fixation in the diaphysis of the native bone, the more complex restoration of joint biomechanics, the extensive soft-tissue resection, the lengthy surgical operations, and the need for chemotherapy and radiation therapy. Because of the difficulties and the high rate of complications associated with the implantation of such highly modular systems, the treatment of patients with malignant bone tumors should only be carried out in specialized centers. PMID- 20001897 TI - Does it make sense that diabetes is reciprocally associated with periodontal disease? AB - Epidemiological studies indicated that more than 15% of the population in western countries suffer because of severe forms of periodontitis, In this respect, the recognition of the relationship between oral and systemic health is growing, thus receiving remarkable interest in scientific literature. In fact, periodontitis may increase the risk for a group of life-threatening conditions such as atherosclerosis, stroke or low birth weight. The American Diabetes Association has reported that individuals with uncontrolled diabetes (defined as 200mg/dL of glucose on three consecutive readings) undergo an increased risk of infections, abnormal wound healing and consequent increased recovery time. Moreover, diabetics may be more likely to develop periodontal and cardiovascular disease than non diabetics, if note. History of poorly controlled chronic periodontal disease can alter diabetic/glycemic control. This may originate from a likely continuous passege of bacterial toxins and/or bacteria into the bloodstream, and/or from an exaggerated release of inflammatory mediators. This review is aimed at elucidating the connections between the status of oral health and glycemic control in diabetes. PMID- 20001898 TI - Relevance of pharmacokinetics and bioavailability of intranasal corticosteroids in allergic rhinitis. AB - Intranasal corticosteroids offer effective symptomatic treatment of allergic rhinitis in children and adults. When used in recommended doses and administration regimens side effect profiles are acceptable and the risk of serious systemic adverse reactions is small. In children, assessments with the sensitive measure of knemometry has found reassuring results. Rigorous comparisons of specific intranasal corticosteroids in clinical use are not available and we have no knowledge as to whether variations in pharmacokinetic properties or bioavailability may reflect significant clinical differences. When the evidence on pharmacokinetics, bioavailability, systemic activity, side effects and efficacy are taken together, however, the most recently launched compounds fluticasone propionate, mometasone furoate, fluticasone furoate and ciclesonide seem preferable to older products. The documented once-daily administration, the quick onset of action and the concurrent effect on eye symptoms of the new drugs support the suggestion. Evidence for intranasal corticosteroids in pre-school children has not been provided. In this paper, a brief review of recent patents for the treatment of allergic rhinitis has also been presented. PMID- 20001899 TI - Signaling pathways associated with inflammatory bowel disease. AB - Inflammatory bowel disease (IBD) consists of Crohn's disease (CD) and ulcerative colitis (UC). It is thought to be caused by genetic, abnormal immune response of the intestinal immune system and dysfunction of intestinal mucosal barrier against enteric bacteria. Mutational genes can affect the development of IBDs via certain signaling pathways. The abnormal signaling pathways play an important role in the inflammatory process and can lead to dysregulation of the inflammatory response and are crucial in the pathogenesis of IBDs. The signaling pathways mainly include P38 MAPK, JNK MAPK, PI3K/Akt, NF-kappaB signaling pathways. Intestinal microorganisms play a key role in the initiation and maintenance of disease. Disorders of signaling pathways including TLR, NF-kappaB can act on the intestinal barrier, and cause uninhibitedly release of effector T cells which are the central cells mediating inflammation in CD. This review highlights relevant patents and a new insight of signaling pathways associated with IBDs will help to develop better therapeutic approaches. PMID- 20001900 TI - Properties of a Kunitz-type trypsin inhibitor from Delonix regia seeds against digestive proteinases of Anagasta kuehniella (Z.) and Corcyra cephalonica (S.) (Lepidoptera: Pyralidae). AB - DrTI was effective against trypsin-like enzymes from A. kuehniella and C. cephalonica, however an artificial diet was insufficient to affect the survival and body weight of either insect. The inhibitor stimulated chymotrypsin-like enzymes and probably induced the synthesis of enzymes insensitive to TLCK in neonate larvae. PMID- 20001901 TI - Two Kunitz-type inhibitors with activity against trypsin and papain from Pithecellobium dumosum seeds: purification, characterization, and activity towards pest insect digestive enzyme. AB - Two trypsin inhibitors (called PdKI-3.1 and PdKI-3.2) were purified from the seeds of the Pithecellobium dumosum tree. Inhibitors were obtained by TCA precipitation, affinity chromatography on Trypsin-Sepharose and reversed-phase HPLC. SDS-PAGE analysis with or without reducing agent showed that they are a single polypeptide chain, and MALDI-TOF analysis determined molecular masses of 19696.96 and 19696.36 Da, respectively. The N-terminal sequence of both inhibitors showed strong identity to the Kunitz family trypsin inhibitors. They were stable over a wide pH (2-9) and temperature (37 to 100 degrees C) range. These inhibitors reduced over 84% of trypsin activity with inhibition constant (Ki) of 4.20 x 10(-8) and 2.88 x 10(-8) M, and also moderately inhibited papain activity, a cysteine proteinase. PdKI-3.1 and PdKI-3.2 mainly inhibited digestive enzymes from Plodia interpunctella, Zabrotes subfasciatus and Ceratitis capitata guts. Results show that both inhibitors are members of the Kunitz-inhibitor family and that they affect the digestive enzyme larvae of diverse orders, indicating a potential insect antifeedant. PMID- 20001903 TI - FCS-based sensing for the detection of ochratoxin and neomycin in food. AB - In this work, we present an advanced fluorescence assay for the detection of traces of ocratoxin A and neomycin in food. The described assay is based on measurement of the fluctuations of the fluorescein-labeled analytes by a focused laser beam in the absence and in the presence of the specific antibodies anti analytes. A competitive assay based on the utilization of unlabeled analytes was developed. The obtained results indicated that the combination of high-avidity IgG antibodies together with an innovative fluorescence immunoassay strategy resulted in the detection limit of 0.0078 ng and 0.0156 ng for ochratoxin A and neomycin, respectively. PMID- 20001904 TI - Identifying sequences potentially related to resistance response of Piper tuberculatum to Fusarium solani f. sp. piperis by suppression subtractive hybridization. AB - Piper tuberculatum is an exotic Piper from the Amazon region that shows resistance to infection by Fusarium solani f. sp. piperis, causal agent of Fusarium disease in black pepper (Piper nigrum L.). In this work we aimed to study the interaction between P. tuberculatum and F. solani f. sp. piperis at a molecular level, using suppression subtractive hybridization to identify genes potentially related to Fusarium disease resistance. Comparative sequence analysis confirmed that clones isolated here show a high identity with genes coding for proteins that have a known role in plant defense response mechanisms, such as peroxidase, hydroxyproline-rich glycoprotein and CBL-interacting protein kinase. The present study constitutes the first effort to understand the molecular basis of this plant-pathogen interaction, identifying genes which may be used in the future genetic improvement of black pepper. PMID- 20001905 TI - Functional cloning and expression of a novel Endo-alpha-1,5-L-arabinanase from a metagenomic library. AB - A novel endo-alpha-L-arabinanase gene (arn2) was isolated, and expressed in E. coli in active form. The recombinant enzyme (ARN2) had optimum activity at pH 6.0 and 45-50( degrees )C with stability between pH 5.0-8.0 and at temperatures up to 40( degrees )C. The recombinant ARN2 catalyzed internal cleavage of alpha-1,5 glycosidic bonds of CM-arabinan, debranched arabinan, linear arabinan, and sugar beet (native) arabinan at rates of decreasing order, and was inactive on wheat arabinoxylan and p-nitrophenyl-alpha-L-arabinofuranoside. Kinetic analysis showed that branching in the arabinan did not significantly affect the apparent K(m) values, and the difference in the reaction rates was likely due to the chemical step after substrate binding. The enzyme hydrolyzed arabino-oligosaccharides of DP> or =6 to smaller oligomers and mostly arabinotriose. Natural and modified arabinans were cleaved to oligomers of various chain lengths, which were progressively hydrolyzed to yield arabinotriose. The pattern of degradation revealed an endo-acting mechanism with arabinotriose as the end product. PMID- 20001906 TI - The small GTPase activity of the ROC domain from LRRK2, a Parkinson's disease related protein. AB - Mutations in the LRRK2 gene have been implicated in the pathogenesis of Parkinson's disease. This work provides biochemical evidence that the ROC domain of LRRK2 functions as a small GTPase, and the Parkinson's disease-associated mutants do not appear to have reduced GTP hydrolysis activities. PMID- 20001907 TI - Robust prediction of B-factor profile from sequence using two-stage SVR based on random forest feature selection. AB - B-factor is highly correlated with protein internal motion, which is used to measure the uncertainty in the position of an atom within a crystal structure. Although the rapid progress of structural biology in recent years makes more accurate protein structures available than ever, with the avalanche of new protein sequences emerging during the post-genomic Era, the gap between the known protein sequences and the known protein structures becomes wider and wider. It is urgent to develop automated methods to predict B-factor profile from the amino acid sequences directly, so as to be able to timely utilize them for basic research. In this article, we propose a novel approach, called PredBF, to predict the real value of B-factor. We firstly extract both global and local features from the protein sequences as well as their evolution information, then the random forests feature selection is applied to rank their importance and the most important features are inputted to a two-stage support vector regression (SVR) for prediction, where the initial predicted outputs from the 1(st) SVR are further inputted to the 2nd layer SVR for final refinement. Our results have revealed that a systematic analysis of the importance of different features makes us have deep insights into the different contributions of features and is very necessary for developing effective B-factor prediction tools. The two-layer SVR prediction model designed in this study further enhanced the robustness of predicting the B-factor profile. As a web server, PredBF is freely available at: http://www.csbio.sjtu.edu.cn/bioinf/PredBF for academic use. PMID- 20001908 TI - A study on the importance of phenylalanine for aurein functionality. AB - Aurein 2.5 (GLFDIVKKVVGAFGSL-NH(2)) is an amphibian antimicrobial peptide. Here, characterisation studies showed the peptide to exhibit molecular areas at an air / water interface (1.77 - 3.1 nm(2)), which are in agreement with the adoption of alpha-helical structures. Lipid monolayer studies showed aurein 2.5 to induce maximal surface pressure changes of circa 7 mN m(-1) in monolayers formed from phosphatidylglycerol (PG) and circa 6 mN m(-1) in those formed from phosphatidylethanolamine (PE). These data indicate that the membrane interactions of the peptide are amphiphilicity driven with no apparent electrostatic requirement. Individually mutating the phenylalanine residues of aurein 2.5 to leucine had no major effect on the levels of PG and PE interactions, suggesting that these residues are not essential to the membrane interactions of the peptide, contrasting to other aureins where corresponding phenylalanine residues are required for efficient membrane interaction and antibacterial activity. This difference in the requirement is suggested to relate to the surface architecture as proposed by the concept of the molecular perturbation potential. PMID- 20001909 TI - N-heterocyclic dipeptide aldehyde calpain inhibitors. AB - A series of Val-Leu based peptidic aldehydes containing either a furan or thiophene at the N-terminus was prepared and assayed against ovine m-calpain. In general, potency is favoured by a 2-substituted (rather than 3-substituted) heterocycle, a thiophene rather than a furan, and a shorter chain length at the N terminus. Molecular docking experiments provide some rationale for these observations. PMID- 20001910 TI - Crystal structure of bucain, a three-fingered toxin from the venom of the Malayan krait (Bungarus candidus). AB - Bucain, a potent neurotoxin isolated from the venom of the Malayan krait (Bungarus candidus), induces paralysis and death. Its crystal structure has been determined at 2.10 A resolution and based on the molecular topology and hydrophobicity profile is structurally classified as a three-fingered alpha neurotoxin possessing a positively charged AChR-binding site. PMID- 20001911 TI - Gpos-mPLoc: a top-down approach to improve the quality of predicting subcellular localization of Gram-positive bacterial proteins. AB - In this paper, a new predictor called "Gpos-mPLoc", is developed for identifying the subcellular localization of Gram positive bacterial proteins by fusing the information of gene ontology, as well as the functional domain information and sequential evolution information. Compared with the old Gpos-PLoc, the new predictor is much more powerful and flexible. Particularly, it also has the capacity to deal with multiple-location proteins as indicated by the character "m" in front of "PLoc" of its name. For a newly-constructed stringent benchmark dataset in which none of included proteins has > 25% pairwise sequence identity to any other in a same subset (location), the overall jackknife success rate achieved by Gpos-mPLoc was 82.2%, which was about 10% higher than the corresponding rate by the Gpos-PLoc. As a user friendly web-server, Gpos-mPLoc is freely accessible at http://www.csbio.sjtu.edu.cn/bioinf/Gpos-multi/. PMID- 20001912 TI - An investigation of the molecular interactions of diacetylcurcumin with ribonuclease A. AB - Curcumin is a natural product with diverse pharmacological activities. Studies of curcumin and its structural derivatives have been a subject of growing interest as a result of their diverse biological activities. We report the interaction of diacetylcurcumin (DAC) with Ribonuclease A (RNase A). The binding constant of DAC with RNase A was found to be of the order of 10(4) M(-1). The intrinsic fluorescence of RNase A was quenched by DAC with a quenching constant of 2.2 x10(4) M(-1). The distance between the fluorophore of RNase A and DAC was found to be 2.6 nm, calculated from a Forster type fluorescence resonance energy transfer (FRET). Secondary structural changes of RNase A after binding were analyzed from circular dichroism and Fourier transform infrared studies. Protein ligand docking studies were conducted to determine the residues involved in the interaction of RNase A with DAC and changes in the accessible surface of the interacting residues were calculated accordingly. PMID- 20001913 TI - The dispersion of water proton spin-lattice relaxation rates in aqueous human protein HC (alpha1-microglobulin) solutions. AB - The (1)H NMR Fast Field Cycling relaxometry was applied to study the molecular dynamics of the human protein HC (alpha (1)-microglobulin), its hydration and aggregation in solution state. The (1)H NMRD data have revealed the complex nature of the water/protein HC system resulting from the co-existence of monomer and dimer forms of the protein in solution as well as the presence of oligosaccharides linked to the polypeptide chain. A comparison of the average correlation time values obtained from the model-free fits with the values predicted on the basis of hydrodynamic tau (r) theory, suggests that the dynamics in solution state is governed mainly by the dimer form of the protein HC (the dominant contribution to the water proton-spin lattice relaxation comes from exchanging protons from the surface of the dimer). The existence of small number of oligomeric forms of the protein HC in solutions is postulated because of the two-step shape of water proton spin-lattice relaxation rate dispersion profiles. PMID- 20001915 TI - Insights on the structure of amyloid fibrils from site-directed mutagenesis. AB - To test the hypothesis that the ability to form ordered beta-rich amyloid fibers with identical structures is a generic property of proteins we present a study on the overall structures of fibers formed by apomyoglobin mutants that either stabilize or destabilize the native state or the intermediate. Our results indicate that, at least at the macroscopic level, ordered beta-rich amyloid fibers have similar structures. PMID- 20001916 TI - Unfoldomics of human genetic diseases: illustrative examples of ordered and intrinsically disordered members of the human diseasome. AB - Intrinsically disordered proteins (IDPs) constitute a recently recognized realm of atypical biologically active proteins that lack stable structure under physiological conditions, but are commonly involved in such crucial cellular processes as regulation, recognition, signaling and control. IDPs are very common among proteins associated with various diseases. Recently, we performed a systematic bioinformatics analysis of the human diseasome, a network that linked the human disease phenome (which includes all the human genetic diseases) with the human disease genome (which contains all the disease-related genes) (Goh, K. I., Cusick, M. E., Valle, D., Childs, B., Vidal, M., and Barabasi, A. L. (2007). The human disease network. Proc. Natl. Acad. Sci. U.S.A. 104, 8685-90). The analysis of this diseasome revealed that IDPs are abundant in proteins linked to human genetic diseases, and that different genetic disease classes varied dramatically in the IDP content (Midic U., Oldfield C.J., Dunker A.K., Obradovic Z., Uversky V.N. (2009) Protein disorder in the human diseasome: Unfoldomics of human genetic diseases. BMC Genomics. In press). Furthermore, many of the genetic disease-related proteins were shown to contain at least one molecular recognition feature, which is a relatively short loosely structured protein region within a mostly disordered segment with the feature gaining structure upon binding to a partner. Finally, alternative splicing was shown to be abundant among the diseasome genes. Based on these observations the human-genetic-disease-associated unfoldome was created. This minireview describes several illustrative examples of ordered and intrinsically disordered members of the human diseasome. PMID- 20001917 TI - Amyloidogenicity and aggregate cytotoxicity of human glucagon-like peptide-1 (hGLP-1). AB - The potential of human glucagon-like peptide-1 (hGLP-1) as a therapeutic agent is limited by its high aggregation propensity. We show that hGLP-1 forms amyloid like structures that are preceded by cytotoxic aggregates, suggesting that aggregation of biopharmaceuticals could present a cytotoxic risk to patients besides the reported increased risk in immunogenicity. PMID- 20001918 TI - Characterization of a saccharide-binding protein from Talisia esculenta seeds with trypsin inhibitory activity. AB - Some proteins exhibit characteristics that suggest they have a primary, if not an exclusive role in nutrient reserve storage. The best studied examples are the storage proteins that accumulate specifically in developing seeds. Some of these protein demonstrate biological activities that could contribute to resistance to pest, pathogens or abiotic stresses. In this study we present the biochemical characterization and cloning of the major protein from seeds of T. esculenta (Talisin), a member of the Sapindaceae family. The N-terminal sequence of the protein isolated was used to produce a degenerated primer. This primer allowed the amplification of the Talisin cDNA by RTPCR from mRNA of the T. esculenta seeds protein. The sequence analysis of the cloned cDNA, demonstrated a 756 bp sequence encoding a peptide of 198 amino acids. The deduced peptide presented high similarity to a typical VSP, the 22-kDa protein in lychee (73 %) and 50.0 % identity to Theobroma bicolor reserve protein. Identities of 52.0 % and 44.0 % to trypsin inhibitors from Treobroma mammosum and Populus tremula respectively. In conclusion, we may suggest that Talisin could be a seed storage protein with affinity properties, i.e. interacts with carbohydrates and trypsin enzyme. PMID- 20001919 TI - Structural and IR-spectroscopic elucidation of dipeptide (L)-threonyl-(L) methionine in solid state. AB - Dipeptide (L)-threonyl-(L)-methionine (Thr-Met) is characterized structurally by means of a solid-state linear polarized IR- spectroscopy (IR-LD) of oriented samples as a colloidal suspension in nematic liquid crystal and quantum chemical ab initio calculations. Vibrational analysis supports the experimental data as well. The role of intermolecular hydrogen bonding on conformational behavior and spectroscopic properties of the compound, studied in solid state is determined. PMID- 20001920 TI - Confocal microscopy evidence of prion protein fragment hPrP[173-195] internalization in rat B104 neuroblastoma cell line. AB - The cytotoxicity of hPrP[173-195] prion peptide against a neuroblastoma cell model was found independent of its tendency to aggregate over time. Cytosolic and nuclear inclusions of peptide were highlighted by confocal microscopy, suggesting a role as a transcription factor in activating signal transduction pathways involved in cell toxicity. PMID- 20001921 TI - Neuronal differentiation of neural progenitor cells by intracellular delivery of synthetic oligopeptide derived from Von Hippel-Lindau protein. AB - Intracellular delivery of synthetic oligopeptides has the potential to promote the occurrence of various cellular events such as cell death, proliferation, growth inhibition, metabolic changes, and morphological changes. However, the regulation of cellular differentiation by intracellular delivery of synthetic oligopeptides has been little studied. Von Hippel-Lindau protein (pVHL) is one of the proteins that functions to induce the differentiation of neural progenitor cells (NPCs). To function in these cells, pVHL forms a complex composed of itself, elongin BC, Clu-2, and Rbx-1. It is suggested that the binding site of elongin BC in pVHL plays a critical role in pVHL function, i.e., ubiquitination, which is related to neuronal differentiation. So, we synthesized an oligopeptide corresponding to the elongin BC binding site, and delivered the oligopeptide into NPCs by using a mixture of trifluoroacetylated lipopolyamine and diloeoyl phosphatidylethanolamine (BioPorter) to form a peptide-lipid complex. After intracellular delivery of the oligopeptide, induction of differentiation of NPCs was shown in terms of neurite outgrowth and by immunocytochemical and electrophysiological means. The intracellular delivery of the synthetic oligopeptide derived from pVHL may provide a safe and valuable approach for the neuronal differentiation of NPCs. PMID- 20001914 TI - The production and role of gastrin-17 and gastrin-17-gly in gastrointestinal cancers. AB - The gastrointestinal peptide hormone gastrin is responsible for initiating the release of gastric acid in the stomach in response to the presence of food and/or humoral factors such as gastrin releasing peptide. However, it has a role in the growth and maintenance of the gastric epithelium, and has been implicated in the formation and growth of gastric cancers. Hypergastrinemia resulting from atrophic gastritis and pernicious anemia leads to hyperplasia and carcinoid formation in rats, and contributes to tumor formation in humans. Additionally, gastrin has been suspected to play a role in the formation and growth of cancers of the colon, but recent studies have instead implicated gastrin processing intermediates, such as gastrin-17-Gly, acting upon a putative, non cholecystokinin receptor. This review summarizes the production and chemical structures of gastrin and of the processing intermediate gastrin-17-Gly, as well as their activities in the gastrointestinal tract, particularly the promotion of colon cancers. PMID- 20001922 TI - Non-globular structures of tandem repeats in proteins. AB - There are two classes of tandem repeats in proteins - globular and non-globular. There are two subclasses of non-globular repeats. The first, such as collagen, form stable helices. Members of the second are flexible and somewhat disordered both in vitro and in vivo. This review focuses on this second subclass. PMID- 20001923 TI - Biochemical and PMF MALDI-TOF analyses of two novel papain-like plant proteinases. AB - Two cysteine endopeptidases from latex of Araujia angustifolia (araujiain aI and araujiain aIII) were purified and characterized by means of conventional and proteomics techniques (MALDI-TOF). N-terminal sequences showed a high percentage of identity with cysteine proteinases belonging to the papain family. The peptide mass fingerprint analysis demonstrated a close homology among both proteinases. PMID- 20001924 TI - Antibodies against refolded recombinant envelope protein (domain III) of Japanese encephalitis virus inhibit the JEV infection to Porcine Stable Kidney cells. AB - Japanese encephalitis virus (JEV) is a mosquito-borne viral zoonosis of public health importance. Global efforts have been made towards development of vaccine for prevention of Japanese encephalitis. The envelope protein of JEV is associated with viral binding to cellular receptors, membrane fusion, and the induction of protective neutralizing antibody response in hosts. Here we report that the antibodies raised against refolded domain III of envelope protein of JEV neutralize the JE virus and inhibit the JEV infection to Porcine Stable Kidney (PS) cells. A reverse transcriptase-PCR amplified gene encoding domain III of JEV envelope protein was cloned into pET28a+ vector and over expressed in E. coli. The recombinant JEV-DIII protein was purified by affinity chromatography under denaturing conditions. The rJEV-DIII was refolded by oxido-redux shuffle and purified to homogeneity by ion-exchange chromatography. Refolded rJEV-DIII was characterized using biochemical and biophysical methods. The polyclonal antibodies were raised against in vitro refolded rJEV-DIII protein in BALB/c mice with Freunds adjuvant. Ninety percent JEV is neutralized when the serum against refolded rJEV-DIII is used at a dilution of 1:80 as against 60.5% neutralization capacity with the same dilution of serum raised against denatured rJEV-DIII. The method of expression and purification of biologically functional rJEV-DIII protein described in this study may help in better understanding the biology of JE virus and the development of better vaccine candidate. Since the expression system uses E. coli as the heterologous host, the process is easy and amenable to inexpensive scale-up. PMID- 20001925 TI - Proteomic analysis of mitochondria in Raji cells following exposure to radiation: implications for radiotherapy response. AB - Radioresistance represents a major obstacle to a successful outcome for the treatment of non-Hodgkin's lymphoma (NHL). Here we performed a global differential proteome analysis of the mitochondria in Raji cells exposed to radiation. The results showed that 23 differentially expressed proteins were identified. Furthermore, GAPDH, RECQL4, MKI67, and ATAD3B could serve as potential biomarkers of radioresistance. PMID- 20001926 TI - Deciphering the flexibility and dynamics of Geobacillus zalihae strain T1 lipase at high temperatures by molecular dynamics simulation. AB - The stability of biocatalysts is an important criterion for a sustainable industrial operation economically. T1 lipase is a thermoalkalophilic enzyme derived from Geobacillus zalihae strain T1 (T1 lipase) that was isolated from palm oil mill effluent (POME) in Malaysia. We report here the results of high temperatures molecular dynamics (MD) simulations of T1 lipase in explicit solvent. We found that the N-terminal moiety of this enzyme was accompanied by a large flexibility and dynamics during temperature-induced unfolding simulations which preceded and followed by clear structural changes in two specific regions; the small domain (consisting of helices alpha3 and alpha5, strands beta1 and beta2, and connecting loops) and the main catalytic domain or core domain (consisting of helices alpha6- alpha9 and connecting loops which located above the active site) of the enzyme. The results suggest that the small domain of model enzyme is a critical region to the thermostability of this organism. PMID- 20001927 TI - Surface Plasmon Resonance Imaging sensor for cathepsin determination based on immobilized cystatin. AB - A specific SPRI sensor for cathepsin determination based on the interaction between immobilized cystatin and cathepsins has been developed. All cathepsins form the same calibration curve. The sensor dynamic response range is between 0.5 and 2.0 ng ml(-1) and the detection limit is equal to 0.1 ng ml(-1). PMID- 20001928 TI - Gibbs sampling shows possibilities of B-cell epitope signatures. AB - We have attempted finding common sequential patterns among protein antigens. For this, we have used Gibbs multiple motif sampler on the set of all non-redundant antigenic sequences available in curated databanks. Several sequential motifs are obtained on these sequences when the amino acids are represented according to their similarity clusters. Significantly high proportions of known B-cell epitope sites are found within or adjacent to these motifs, thus indicating a possibility of linear epitope signatures. These findings may offer important applications in synthesis of peptide vaccines. A predictive example in this regard is presented. PMID- 20001929 TI - Dronedarone--a new alternative for management of atrial fibrillation. AB - The maintenance of sinus rhythm in patients with Atrial Fibrillation (AF) is difficult and is complicated by adverse drug reactions of antiarrhythmic drugs or the adverse events related with ablation procedures. Amiodarone can be used for preventing AF recurrence; but it has the risk of serious toxicities secondary to its iodinated nature. Dronedarone, an amiodarone analogue without iodine, has been shown to decrease the frequency of AF recurrence and to provide heart rate control during AF recurrence. Dronedarone is a benzofuran derivative; specifically N-[2-Butyl-3-[4-[3-(dibutylamino)propoxy]benzoyl]-5 benzofuranyl]methane-sulfonamide and its production process has been patented. In the ATHENA trial, dronedarone therapy was associated with a significant reduction in the combined primary endpoint of death from any cause or hospitalization due to cardiovascular causes. However, dronedarone has been noted to increase cardiovascular mortality among patients with advanced congestive heart failure (NYHA class III and IV). On March 18, 2009, the Cardiovascular and Renal Drugs Advisory Committee of the Food and Drug Administration (FDA) approved dronedarone for an indication of atrial fibrillation suppression, with a warning against its use in patients with NYHA class III and IV heart failure or recently decompensated heart failure. PMID- 20001930 TI - Functional outcome in patients treated for chronic posttraumatic osteomyelitis. AB - BACKGROUND: Management of chronic posttraumatic osteomyelitis remains an important challenge in orthopaedics. In this investigation, 43 patients at a large university hospital were retrospectively identified who had been diagnosed with chronic osteomyelitis at 44 sites. METHODS: Diagnosis was based on the patient's history and physical examination, laboratory values, radiological evaluation, and intraoperative specimens. Patients' charts were reviewed and follow-up was obtained on 33 patients in whom Short Musculoskeletal Functional Assessment (SMFA) questionnaires were obtained. RESULTS: The mean follow-up of the cohort was 21.1 months (range, 10 to 54 months). Sites involved included: tibia (55%) and femur (36%), with the remainder involving the pelvis, radius/ulna, and calcaneus. Blood, bone, or wound aspirate cultures were positive in 85%, most commonly for Staphylococcus aureus. 84% of the patients in this investigation were completely cured after a single surgical intervention, removal of hardware and a course of intravenous antibiotics. Three patients required additional surgical intervention and two remained infected. Ultimately, the cohort reached a cure rate of 94%. SMFA data revealed that patients had a dysfunction score of 53.8 and bother index of 51.5, with 50 being the normal for the general population. CONCLUSION: The patients in this cohort functioned at a level less than one standard deviation below the general population. These results suggest that patients with chronic posttraumatic osteomyelitis do not function significantly below that of the normal population following eradication of their infections. PMID- 20001931 TI - The medial malleolus osteoligamentous complex and its role in ankle fractures. AB - Ankle stability in ankle fractures is dependent on multiple factors. The medial malleolus and the associated deltoid ligament provide for ankle stability on the medial side. Over the years, the relative importance of this medial malleolar osteoligamentous complex (MMOLC) has been debated. This review will describe the evolution of ankle fracture surgery from the perspective of the contribution of the MMOLC to re-establishing ankle stability. Also discussed are the surgical and nonsurgical treatment options, various presentations of medial sided injuries in ankle fractures, and, finally, current recommendations for fixation. PMID- 20001932 TI - Physiological knee laxity during pubertal growth. AB - BACKGROUND: The relationship of physiologic laxity to age has been reported in only cross-sectional studies. The current investigators suggest further investigations, to include Tanner staging in order to understand the extent to which increases in maturity influence alterations in laxity. MATERIALS AND METHODS: A two-phase (cross-sectional and longitudinal) study assessed knee joint laxity and flexibility in 172 normal adolescents, using a KT 2000 arthrometer, anthropometric measurements, and Carter and Wilkinson tests. Correlation of these evaluations was done with gender and Tanner stage. Data from clinical and KT 2000 measurements at the initial evaluation were analyzed as the cross-sectional, single assessment phase of the study. The longitudinal phase of the study reviewed modulation of laxity and flexibility, with growth by repeated examination of the above tests in individual subjects during their adolescent growth phase. RESULTS: Increased flexibility was seen significantly more frequently in females than males in both study phases. Age, Tanner stage, and anthropometric values were not significantly associated with laxity in the cross sectional study. In the longitudinal study, an inverse relationship was demonstrated between Tanner stage and KT 2000 laxity measures after adjusting for other variables. Sequential evaluation showed a progressive decrease of sagittal laxity at the onset of Tanner stage 2. Laxity was significantly greater in adolescents, with signs of joint physiologic hyperflexibility. CONCLUSION: Evaluation of laxity and flexibility during the adolescent growth phase is important for better definition of muscle strengthening or flexibility programs, to avoid functional overloads and injury. PMID- 20001933 TI - Arthroscopic management of calcific tendinopathy of the shoulder--do we need to remove all the deposit? AB - Results of arthroscopic management of chronic, recalcitrant calcific tendinopathy of the shoulder in 28 patients were evaluated. Twenty-six patients (average age, 44 years) were ultimately enrolled in the study. Postoperative radiographs were performed to determine the amount of calcium deposit removal. Radiographic examination 2 months after surgery was performed to evaluate further deposit changes. The Constant score showed a statistically significant improvement in all patients. Better results were obtained when complete removal of the calcifications was achieved. On radiographic examination, performed two months after surgery, no calcification deposit increased in size and no translucent deposit changed into a dense deposit. Most of the dense deposits, partially removed, changed into translucent deposits. In this series, we found arthroscopic management was effective in treating calcific tendinopathy. PMID- 20001934 TI - Can the Bio-Transfx pin fail during initial ACL graft insertion? AB - BACKGROUND: The Bio-Transfix pin is a biodegradable device used for femoral tunnel anterior cruciate ligament (ACL) graft fixation. Recent clinical studies have suggested the possibility of the pin's postoperative failure. METHODS: This investigation evaluates the initial strength of several Bio-Transfix pin ACL fixations in a simulated femoral tunnel model. The forces generated by five surgeons during simulated ACL graft tensioning were also measured. RESULTS: Average strengths of the pins ranged from 1075 to 2160 N for 10 and 8 mm tunnels, respectively, whereas the maximum surgeon-generated forces were 535 N. CONCLUSIONS: These results imply that initial fracture of the pin itself is unlikely; however, failure of the supporting bone or a decrease in pin strength due to biodegradation could account for early loss of the fixation. PMID- 20001935 TI - Proximal biceps tendon--a biomechanical analysis of the stability at the bicipital groove. AB - The subscapularis tendon, coracohumeral ligament, and transverse humeral ligament are all believed to contribute to biceps tendon stability within the bicipital groove. In order to examine the relative contribution of these soft tissue structures to proximal biceps tendon stability, 11 fresh frozen cadaveric shoulder specimens were prepared and mounted onto a custom jig. A three dimensional digitizer was utilized to record biceps tendon excursion in various shoulder positions. In sequential order, these structures were then sectioned, and biceps tendon excursion was again recorded. We found that sectioning of the subscapularis tendon significantly increased biceps tendon excursion, compared to intact specimens (8.1 +/- 4.1 mm vs. 4.3 +/- 3.6 mm; p < 0.006). In contrast, isolated sectioning of the transverse humeral ligament or the coracohumeral ligament did not significantly increase biceps excursion (5.4 +/- 2.5 mm, p = 0.26; 5.6 +/- 1.3 mm, p = 0.24). When two structures were sectioned, significant excursion in the biceps tendon only occurred in specimens where the subscapularis tendon was one of the sectioned structures. The preliminary data suggest that, of the three tested soft tissue structures, the subscapularis tendon is the most important stabilizer of the proximal biceps and that clinically significant lesions of the proximal biceps tendon may be associated with a defect in the subscapularis tendon. PMID- 20001936 TI - Femoral component positioning in resurfacing arthroplasty--effects on cortical strains. AB - Previous studies have suggested that femoral component positioning in resurfacing arthroplasty may affect strains in the femoral neck that could lead to decreased implant longevity. A strain gaged, Sawbones model was used to determine the femoral neck strains for a variety of resurfacing head translations and angulations. We found that head positions affected strain distributions, most positions leading to increased neck strains, often over 100%, with the exception being a varus head position where the superior neck strains decreased over 50%. Although the clinical meaning of these findings is unclear, it could be of concern for stress-shielding or fatigue fracture of the femoral neck. PMID- 20001937 TI - Long-term functional outcome and donor-site morbidity associated with autogenous iliac crest bone grafts utilizing a modified anterior approach. AB - Prior studies and techniques for harvesting iliac crest bone have shown significant postoperative pain, disability, and poor cosmesis. This retrospective study was conducted to examine bone graft donor-site morbidity by evaluating functional outcomes in patients who have undergone a modified anterior harvesting approach. The medical charts and hospital records of 43 patients were retrospectively reviewed over a 6-year period. Demographic information, operative notes, laboratory results, and the American Society of Anesthesiologists (ASA) classification were recorded. All patients were evaluated retrospectively at a mean 41 months after bone-graft harvesting. Patients available for follow-up were asked to quantify their pain level at the donor-site on a visual analog pain scale (0-10). They also completed SMFA forms, as well as a survey pertaining to sensory deficits, gait disturbances, and cosmetic appearance. Forty-four patients met the inclusion criteria consisting of 25 males and 18 females, mean age 47 years (range, 22 to 80 years). A total of 32 (73%) patients were available for long-term follow-up at a mean of 41.3 months (range, 8 to 83 months). Eight (25%) of these patients reported minimal postoperative pain at time of follow-up. Three of 32 (9%) patients reported minor ambulation difficulty as a result of donor site pain. Other minor complications included hypertrophic scar formation (7%) and hematoma/seroma (3%). There were no major complications reported, such as deformity at the crest site (0%) or infection (0%). SMFA scores demonstrated a mean dysfunction score of 48.5 (range, 41.8 to 71.1) and a bother index of mean 47.9 (range, 42.6 to 73.9). Utilizing the anterior approach in iliac crest bone harvesting provides an abundant supply of both cortical and cancellous bone, an aesthetically favorable scar, and decreased postoperative donor-site pain. There were very few complications seen in our cohort as compared to previous studies with very good long-term functional outcomes. PMID- 20001938 TI - Chondroprotective effect of N-acetylglucosamine and hyaluronate in early stages of osteoarthritis--an experimental study in rabbits. AB - Osteoarthritis, the most common joint disease in the world, is characterized by joint pain, stiffness, and limitation of range of motion. Osteoarthritis is a slowly progressive disease and its morbidity increases with age. The most commonly involved sites are the spine, knee, hip, and hand joints. Although the ideal treatment for osteoarthritis should be the one that acts on the underlying mechanism, thus preventing joint destruction and disease progression, such an effective treatment option does not exist. Therefore, contemporary treatment aims to relieve pain, increase range of motion, and optimize joint function. Analgesics and nonsteroidal antiinflammatory drugs are most commonly used for the symptomatic treatment, but mainly their gastrointestinal side effects, especially in elderly patients, limit their usage. In this study, the chondroprotective effects of an aminomonosaccharide glucosamine and a polysaccharide hyaluronic acid in a rabbit osteoarthritis model were investigated. Anterior cruciate ligament transection was performed in 32 New Zealand rabbits to establish a model of osteoarthritis. Rabbits were randomized into four groups, each consisting of eight rabbits. Two weeks after the operation, intraarticular injections were performed to the right knees once a week for 5 weeks; intraarticular glucosamine to the first group, intraarticular hyaluronate to the second group, intraarticular hyaluronate and intramuscular glucosamine to the third group, and intraarticular saline solution to the fourth group, which served as the control group. At the end of the eighth week, the rabbits were sacrificed and their right knees with proximal femur and distal tibia were harvested. Joint surfaces of their femur and tibia were examined macroscopically, and sections from the medial femoral condyles were examined microscopically. Macroscopic evaluation revealed that the cartilage surface was preserved in the glucosamine, hyaluronate, and hyaluronate plus glucosamine groups, when compared with the control group. Microscopic evaluation showed that glucosamine, hyaluronate, and glucosamine plus hyaluronate have chondroprotective effect, but no statistically significant difference was found between study groups. PMID- 20001939 TI - Drug-induced myopathies. AB - The most common drugs currently in use that may cause myopathies were reviewed using the Medline database (U.S. National Library of Medicine, Bethesda, Maryland). Our review included results from epidemiologic and database surveys, clinical trials, and case reports. The clinical spectrum is wide, and presentations range from asymptomatic elevations in serum creatine phosphokinase levels to severe life-threatening rhabdomyolysis. Management of suspected drug induced myopathy should include immediate discontinuation of the offending agent, as well as supportive care when needed. Earlier diagnosis and drug discontinuation raises the likelihood of resolution and recovery. PMID- 20001940 TI - Adult periarticular locking plates for the treatment of pediatric and adolescent subtrochanteric hip fractures. AB - Two cases are presented in which adult, precontoured, lower-extremity periarticular locking plates were utilized for fixation of subtrochanteric femur fractures in pediatric patients. Recognition of the fact that a distal tibial locking plate in a small child and a proximal tibial locking plate in an adolescent anatomically ft the proximal femur in each case may provide a surgeon treating subtrochanteric hip fractures in this population increased options for operative stabilization. PMID- 20001941 TI - Peroneal nerve injury with foot drop complicating ankle sprain--a series of four cases with review of the literature. AB - Foot drop has many etiologies. One rarely mentioned and often neglected reason for foot drop is an acute inversion sprain of the ankle. Over the past 14 years, a collection of 32 cases of foot drop have been compiled in our orthopaedic and physiatric practices. All cases had appropriate evaluations, including electrodiagnostic studies (electromyography and nerve conduction studies) to determine the location and type of injury. Treatment and follow-up are also discussed. Of the 32 case studies, four were caused by a straightforward acute inversion sprain of the ankle. These cases are described with the electrodiagnostic evaluations, treatments, and outcomes. Proposed mechanisms for this type of foot drop are discussed, including traction and compression of the common peroneal nerve as it winds around the neck of the fibula, and possible compression by hematoma. Surgical versus conservative treatment is described. The functional impairment associated with foot drop is detailed. PMID- 20001942 TI - Bilateral glenohumeral septic arthritis secondary to retroperitoneal abscess. AB - Glenohumeral septic arthritis is rare and usually a result of Staphylococcus aureus infection. Gram-negative septic arthritis is on the increase and is usually associated with intraabdominal pathology. We present a case of bilateral E. Coli glenohumeral septic arthritis associated with retroperitoneal abscess and discuss pitfalls in diagnosis and management. PMID- 20001943 TI - Obturator dislocation of total hip arthroplasty--a case report. AB - This study consists of a single case report of a patient who had an irreducible obturator dislocation of a total hip arthroplasty after a motor vehicle accident, not previously described in the English literature. In particular, the focus will be on offering an educated opinion on the risk factors for dislocation and difficulties encountered with this type of dislocation. The aim is to offer valuable insight based on the operative experience with this patient and to supplement the literature with the management of such a complication following total hip arthroplasty. PMID- 20001944 TI - Use of an alumina ceramic-on-alumina ceramic bearing surface in THA in a 13 year old with JIA--a single case study. AB - Total hip arthroplasty (THA) has been shown to be highly effective in ameliorating pain and increasing function in adults with end stage arthritis of the hip. Early studies of cemented THA in children with advanced hip disease reported poor results, but, more recently, results of cementless THA show greatly improved outcomes. More recently, concerns related to THA in children have focused on wear of the bearing surfaces, which may result in periprosthetic bone loss and eventual loosening of the prosthetic components. This case report describes the use of an alumina ceramic-on-alumina ceramic bearing surface in THA in a 13-year-old female with bilateral end-stage arthritis of the hips. At 7 and 8 years post-THA, the patient has no pain, no limp, and is able to walk long distances without difficulty. Radiographs show no signs of implant loosening, osteolysis, or wear of the bearing surface. PMID- 20001945 TI - Isolated rupture of the popliteus muscle with painful ossification in a skeletally immature athlete--a case report. AB - A case of an isolated popliteus tendon rupture occurring during sport in a skeletally immature athlete is presented. Treatment is not always clearly defined, as both nonoperative and operative have been successful. Because the outcome of rest from sports activity and failed trials of physical therapy allowed continued discomfort in the posterolateral aspect of the knee in this patient, repeat imaging and arthroscopy were performed. Part of the popliteus tendon was demonstrated to have ossified and was openly debrided. The remaining tendon was repaired to the lateral capsule. The patient went on to a full recovery and return to sport. PMID- 20001946 TI - Lupus cerebritis after visiting a tanning salon. AB - A 53-year-old female was admitted to the intensive care unit for lupus cerebritis; she had a 15-year history of stable lupus. Over the prior 1 to 2 months, the patient visited a tanning salon and this triggered the exacerbation of lupus. Her initial symptoms were cutaneous in the form of an erythematous rash. Within 2 weeks she started to have headaches and was admitted for seizure and psychosis. Ultraviolet A exposure in the tanning salon is known to exacerbate lupus by modulation of the immune system at the level of the skin. It has also been found that ultraviolet light can lead to the formation of antinuclear antibodies. This case illustrates the need to emphasize the danger of the tanning salon to patients with systemic lupus erythematous; the risk is not only cutaneous, it can also be systemic. PMID- 20001947 TI - Seeing solid: realistic depth discerned in single images. PMID- 20001948 TI - The medicare fee schedule was not evolved to be a basis for pediatric medical care. PMID- 20001949 TI - Impairment of a normal eye by a bad eye thru' binocular vision. improvements in strabismus surgery. Moral hazard. An elected and immune Villain. PMID- 20001950 TI - Is one eye better than two in strabismus? Or does the misaligned amblyopic eye interfere with binocular vision? A preliminary functional MRI study. AB - PURPOSE: The aim of this study was to determine if patients with strabismic amblyopia could have increased occipital visual cortex activation with monocular stimulation of the sound fixing eye, rather than with simultaneous stimulation of both eyes. METHODS: A prospective study was performed including 12 patients with strabismus and amblyopia, who were evaluated using functional MRI with visual stimulation paradigms. The measurements were made in the occipital visual cortex, assessing the response to the binocular and monocular stimulation. RESULTS: 12 out of 12 patients showed an increased cortical response of the healthy eye in comparison to the amblyopic one. Nine of the 12 patients showed larger cortical activation with visual stimulation of the healthy eye compared to the binocular condition analysis. Three out of the 12 cases had a greater activation area when the stimulation was binocular rather than monocular, 2 of whom had a relatively small angle of strabismus. CONCLUSIONS: Patients with amblyopia and strabismus could see better with only one eye instead of both eyes. This could be related to inhibition of the binocular function of the brain by the misaligned amblyopic eye. PMID- 20001951 TI - Further implications of probable changes in medial rectus muscle innervation after surgery for infantile esotropia. AB - INTRODUCTION: The conclusion that setting the eyes of an infantile esotrope (ET) straight with a successful bilateral medial rectus muscle reseccion (MROU) reduces the preoperative hyperinnervation of these muscles almost to normal may have special implications for the best principles of management of infantile ET not detailed in a recent paper by Mimms, III, Miller and Schoolfield. METHODS: The same 113 infantile esotropes who provided data for the exoshift under anesthesia study had previously provided data for a dose-response curve. A simple geometric calculation was done to determine the amount of medial rectus (MR) recession necessary to compensate for contracture, and this was substracted from the dose-response value to reveal the additional mm of recession required to the MR sufficiently down the length-tension curve to compensate for the preoperative MR hyperinnervation. RESULTS: The result of these calculations yielded a surprisingly narrow range of averaging 3.6 mm (range 3.2 to 3.8 mm) for a broad range of preoperative deviations (20 ET to 80 ET). CONCLUSIONS: The conclusions which may be drawn from this analysis include an understanding of why infantiles (and other) esotropias tend to increase in size in a few weeks or months if unteated, verification of the wisdom of the common practice of measuring the deviation no more than a day or two prior to the surgery, and the common observation that unilateral medial rectus recessions of a given amount will generally produce significantly less than half of the effect of a similarly-sized bilateral medial rectus recession. Finally, this understanding explains the poor success rate of surgery in the presence of dense amblyopia with eccentric fixation. PMID- 20001952 TI - Cul-de-sac incision for strabismus surgery in older patients. AB - INTRODUCTION: A limbal surgical approach for strabismus surgery is often recommended for older patients, in the belief that the conjunctiva of older patients is too thin and prone to tear during surgery performed through a cul-de sac incision. The purpose of this study was to evaluate the use of the cul-de-sac approach in patients more than 40 years of age. METHODS: This is a retrospective analysis of consecutive patients more than 40 years of age who underwent strabismus surgery using a modified cul-de-sac approach to reduce manipulation of the conjunctiva during surgery. RESULTS: A total of 32 cul-de-sac incisions were used to operate on 37 muscles in 19 consecutive patients more than 40 years old. The man age was 58 yeasr (41-77 years) with 8 patients (12 incisions) more than 60 years old. Absorbable sutures were used to close 28 of the incisions, with 19 incisions requiring 1 suture, 7 requiring 2 sutures, and 2 incisions in 2 patients requiring more than 2 sutures. Tears resulting in extension of the conjunctival incision occurred in several patients, but no other complications were encountered. One week following surgery, 28 (87.5%) of the incisions were well closed, while a small opening was noted with 4 (12.5%) of the incisions, but none required further intervention, and all healed well. CONCLUSION: Strabismus surgery can be performed through a cul-de-sac incision in older patients. Slight modifications of the surgical technique to prevent excessive manipulation of the incision and to reduce the risk of tearing the conjunctiva are helpful. The cul de-sac approach offers some important potential advantages to older patients undergoing strabismus surgery. PMID- 20001954 TI - Proteomic identification and characterization of secreted N-glycosylated NPC2 following cross-linking of the high-affinity receptor for IgE on mast cells. AB - Allergen-mediated cross-linking of the high-affinity receptor for IgE on mast cells triggers the release of diverse preformed and de novo synthesized immunoregulatory mediators that further the allergic response. A proteomic screen applied to the detection of proteins secreted by the model rat mast cell line, RBL-2H3 (rat basophilic leukaemia, subline 2H3.1), led to the identification of the cholesterol-binding glycoprotein, NPC2/RE1 (Niemann-Pick Type C2/epididymal secretory protein 1). Glycosylated NPC2 is secreted early in response to an IgE mediated stimulus and co-localizes with the lysosomal membrane marker, CD63. NPC2 belongs to the ML (MD-2-related lipid-recognition) protein family (155 members), which includes the Toll-like receptor co-factors, MD-1 and MD-2, and perhaps most interestingly, seven major house dust mite allergens of unknown function (including Der p 2 and Der f 2). Possible role(s) for the protein in the allergic response and future applications of this approach are discussed. PMID- 20001953 TI - Regulation of oligodendrocyte progenitor cell maturation by PPARdelta: effects on bone morphogenetic proteins. AB - In EAE (experimental autoimmune encephalomyelitis), agonists of PPARs (peroxisome proliferator-activated receptors) provide clinical benefit and reduce damage. In contrast with PPARgamma, agonists of PPARdelta are more effective when given at later stages of EAE and increase myelin gene expression, suggesting effects on OL (oligodendrocyte) maturation. In the present study we examined effects of the PPARdelta agonist GW0742 on OPCs (OL progenitor cells), and tested whether the effects involve modulation of BMPs (bone morphogenetic proteins). We show that effects of GW0742 are mediated through PPARdelta since no amelioration of EAE clinical scores was observed in PPARdelta-null mice. In OPCs derived from E13 mice (where E is embryonic day), GW0742, but not the PPARgamma agonist pioglitazone, increased the number of myelin-producing OLs. This was due to activation of PPARdelta since process formation was reduced in PPARdelta-null compared with wild-type OPCs. In both OPCs and enriched astrocyte cultures, GW0742 increased noggin protein expression; however, noggin mRNA was only increased in astrocytes. In contrast, GW0742 reduced BMP2 and BMP4 mRNA levels in OPCs, with lesser effects in astrocytes. These findings demonstrate that PPARdelta plays a role in OPC maturation, mediated, in part, by regulation of BMP and BMP antagonists. PMID- 20001955 TI - Complexities of oestrogen in stroke. AB - Evidence exists for the potential protective effects of circulating ovarian hormones in stroke, and oestrogen reduces brain damage in animal ischaemia models. However, a recent clinical trial indicated that HRT (hormone-replacement therapy) increased the incidence of stroke in post-menopausal women, and detrimental effects of oestrogen on stroke outcome have been identified in a meta analysis of HRT trials and in pre-clinical research studies. Therefore oestrogen is not an agent that can be promoted as a potential stroke therapy. Many published reviews have reported the neuroprotective effects of oestrogen in stroke, but have failed to include information on the detrimental effects. This issue is addressed in the present review, along with potential mechanisms of action, and the translational capacity of pre-clinical research. PMID- 20001956 TI - STIM and Orai proteins: players in sexual differences in hypertension-associated vascular dysfunction? AB - Sex-associated differences in hypertension have been observed repeatedly in epidemiological studies; however, the mechanisms conferring vascular protection to females are not totally elucidated. Sex-related differences in intracellular Ca(2+) handling or, more specifically, in mechanisms that regulate Ca(2+) entry into vascular smooth muscle cells have been identified as players in sex-related differences in hypertension-associated vascular dysfunction. Recently, new signalling components that regulate Ca(2+) influx, in conditions of intracellular store depletion, were identified: STIM1 (stromal interaction molecule 1), which works as an intracellular Ca(2+) sensor; and Orai1, which is a component of the CRAC (Ca(2+) release-activated Ca(2+)) channels. Together, these proteins reconstitute store-operated Ca(2+) channel function. Disturbances in STIM1/Orai1 signalling have been implicated in pathophysiological conditions, including hypertension. In the present article, we analyse evidence for sex-related differences in Ca(2+) handling and propose a new hypothesis where sex-related differences in STIM/Orai signalling may contribute to hypertension-associated vascular differences between male and female subjects. PMID- 20001957 TI - Caught in the middle: the role of Bag3 in disease. AB - Bag3 is a Bag family co-chaperone that regulates the ATPase activity of Hsp70 (heat-shock protein 70) chaperones. Recent studies have demonstrated that Bag3 can initiate macroautophagy in co-operation with small heat-shock protein HspB8. In this issue of the Biochemical Journal, Fuchs and co-workers have discovered the IPV motif in Bag3 that is necessary for binding to HspB8. The authors have also identified HspB6 as a new binding partner for Bag3 and characterized further the binding of both HspB8 and HspB6 in Bag3-mediated clearance of aggregated polyglutamine-containing protein Htt43Q (huntingtin exon 1 fragment with 43 CAG repeats). It is clear from recent identification of a Bag3 mutation that causes a form of muscular dystrophy that the full function of Bag3 in disease is not clear. We will apply the findings of Fuchs et al. in this issue to reconcile the phenotypes of Bag3 homologue knockouts with the emerging role of Bag3 in autophagy. PMID- 20001958 TI - 'Unknown' proteins and 'orphan' enzymes: the missing half of the engineering parts list--and how to find it. AB - Like other forms of engineering, metabolic engineering requires knowledge of the components (the 'parts list') of the target system. Lack of such knowledge impairs both rational engineering design and diagnosis of the reasons for failures; it also poses problems for the related field of metabolic reconstruction, which uses a cell's parts list to recreate its metabolic activities in silico. Despite spectacular progress in genome sequencing, the parts lists for most organisms that we seek to manipulate remain highly incomplete, due to the dual problem of 'unknown' proteins and 'orphan' enzymes. The former are all the proteins deduced from genome sequence that have no known function, and the latter are all the enzymes described in the literature (and often catalogued in the EC database) for which no corresponding gene has been reported. Unknown proteins constitute up to about half of the proteins in prokaryotic genomes, and much more than this in higher plants and animals. Orphan enzymes make up more than a third of the EC database. Attacking the 'missing parts list' problem is accordingly one of the great challenges for post-genomic biology, and a tremendous opportunity to discover new facets of life's machinery. Success will require a co-ordinated community-wide attack, sustained over years. In this attack, comparative genomics is probably the single most effective strategy, for it can reliably predict functions for unknown proteins and genes for orphan enzymes. Furthermore, it is cost-efficient and increasingly straightforward to deploy owing to a proliferation of databases and associated tools. PMID- 20001959 TI - Molecular mechanisms of metabolic regulation by insulin in Drosophila. AB - The insulin signalling pathway is highly conserved from mammals to Drosophila. Insulin signalling in the fly, as in mammals, regulates a number of physiological functions, including carbohydrate and lipid metabolism, tissue growth and longevity. In the present review, I discuss the molecular mechanisms by which insulin signalling regulates metabolism in Drosophila, comparing and contrasting with the mammalian system. I discuss both the intracellular signalling network, as well as the communication between organs in the fly. PMID- 20001960 TI - Breaking the code: Ca2+ sensors in plant signalling. AB - Ca2+ ions play a vital role as second messengers in plant cells during various developmental processes and in response to environmental stimuli. Plants have evolved a diversity of unique proteins that bind Ca2+ using the evolutionarily conserved EF-hand motif. The currently held hypothesis is that these proteins function as Ca2+ sensors by undergoing conformational changes in response to Ca2+ binding that facilitate their regulation of target proteins and thereby co ordinate various signalling pathways. The three main classes of these EF-hand Ca2+sensors in plants are CaMs [calmodulins; including CMLs (CaM-like proteins)], CDPKs (calcium-dependent protein kinases) and CBLs (calcineurin B-like proteins). In the plant species examined to date, each of these classes is represented by a large family of proteins, most of which have not been characterized biochemically and whose physiological roles remain unclear. In the present review, we discuss recent advances in research on CaMs and CMLs, CDPKs and CBLs, and we attempt to integrate the current knowledge on the different sensor classes into common physiological themes. PMID- 20001961 TI - Transcriptional and post-translational regulation of adiponectin. AB - Adiponectin is an adipose-tissue-derived hormone with anti-diabetic, anti atherogenic and anti-inflammatory functions. Adiponectin circulates in the bloodstream in trimeric, hexameric and high-molecular-mass species, and different forms of adiponectin have been found to play distinct roles in the regulation of energy homoeostasis. The serum levels of adiponectin are negatively correlated with obesity and insulin resistance, yet the underlying mechanisms remain elusive. In the present review, we summarize recent progress made on the mechanisms regulating adiponectin gene transcription, multimerization and secretion. We also discuss the potential relevance of these studies to the development of new clinical therapy for insulin resistance, Type 2 diabetes and other obesity-related metabolic disorders. PMID- 20001962 TI - Guidelines for the effective use of chemical inhibitors of protein function to understand their roles in cell regulation. AB - Cell-permeable chemical inhibitors have become invaluable reagents for the study of cellular regulation. In the present paper, using protein kinase inhibitors as the example, a set of criteria are described that should be met before any investigation using such compounds should be accepted for publication. PMID- 20001963 TI - Caspase cleavage of Ets-1 p51 generates fragments with transcriptional dominant negative function. AB - Ets-1 is a transcription factor that plays an important role in various physiological and pathological processes, such as development, angiogenesis, apoptosis and tumour invasion. In the present study, we have demonstrated that Ets-1 p51, but not the spliced variant Ets-1 p42, is processed in a caspase dependent manner in Jurkat T-leukaemia cells undergoing apoptosis, resulting in three C-terminal fragments Cp20, Cp17 and Cp14 and a N-terminal fragment, Np36. In vitro cleavage of Ets-1 p51 by caspase 3 produces fragments consistent with those observed in cells undergoing apoptosis. These fragments are generated by cleavage at three sites located in the exon VII-encoded region of Ets-1 p51. This region is absent from the Ets-1 p42 isoform, which therefore cannot be cleaved by caspases. In Ets-1 p51, cleavage generates C-terminal fragments containing the DNA-binding domain, but lacking the transactivation domain. The Cp17 fragment, the major cleavage product generated during apoptosis, is devoid of transcriptional activity and inhibits Ets-1 p51-mediated transactivation of target genes by competing with Ets-1 p51 for binding to Ets-binding sites present in the target promoters. In the present study, we have demonstrated that caspase cleavage of Ets-1 within the exon VII-encoded region leads to specific down regulation of the Ets-1 p51 isoform during apoptosis. Furthermore, our results establish that caspase cleavage generates a stable C-terminal fragment that acts as a natural dominant-negative form of the full-length Ets-1 p51 protein. PMID- 20001964 TI - Glucose induces expression of rat pyruvate carboxylase through a carbohydrate response element in the distal gene promoter. AB - Pyruvate carboxylase is an enzyme of the so-called pyruvate cycling pathways, which have been proposed to contribute to glucose-stimulated insulin secretion in pancreatic beta-cells. In the rat insulinoma cell line 832/13, transcripts from both the distal and proximal gene promoter for pyruvate carboxylase are up regulated by glucose, with pyruvate carboxylase being expressed mainly from the distal gene promoter. At position -408 to -392 relative to the transcription start site, the distal gene promoter was found to contain a ChoRE (carbohydrate response element). Its deletion abolishes glucose responsiveness of the promoter, and the sequence can mediate glucose responsiveness to a heterologous gene promoter. ChREBP (carbohydrate response element-binding protein) and its dimerization partner Mlx (Max-like protein X) bind to the ChoRE in vitro. ChREBP further binds to the distal promoter region at a high glucose concentration in situ. The E-box-binding transcription factors USF1/2 (upstream stimulatory factor 1/2) and E2A variant 2 [also known as E47 and TCF3 (transcription factor 3)] can also bind to the ChoRE. Overexpression of E2A diminishes the magnitude of the glucose response from the pyruvate carboxylase ChoRE. This illustrates that competition between ChREBP-Mlx and other factors binding to the ChoRE affects glucose responsiveness. We conclude that a ChoRE in the distal gene promoter contributes to the glucose-mediated expression of pyruvate carboxylase. PMID- 20001965 TI - Post-transcriptional regulation of MEK-1 by polyamines through the RNA-binding protein HuR modulating intestinal epithelial apoptosis. AB - MEK-1 [MAPK (mitogen-activated protein kinase) kinase-1] is an important signal transducing enzyme that is implicated in many aspects of cellular functions. In the present paper, we report that cellular polyamines regulate MEK-1 expression at the post-transcriptional level through the RNA-binding protein HuR (Hu-antigen R) in IECs (intestinal epithelial cells). Decreasing the levels of cellular polyamines by inhibiting ODC (ornithine decarboxylase) stabilized MEK-1 mRNA and promoted its translation through enhancement of the interaction between HuR and the 3'-untranslated region of MEK-1 mRNA, whereas increasing polyamine levels by ectopic ODC overexpression destabilized the MEK-1 transcript and repressed its translation by reducing the abundance of HuR-MEK-1 mRNA complex; neither intervention changed MEK-1 gene transcription via its promoter. HuR silencing rendered the MEK-1 mRNA unstable and inhibited its translation, thus preventing increases in MEK-1 mRNA and protein in polyamine-deficient cells. Conversely, HuR overexpression increased MEK-1 mRNA stability and promoted its translation. Inhibition of MEK-1 expression by MEK-1 silencing or HuR silencing prevented the increased resistance of polyamine-deficient cells to apoptosis. Moreover, HuR overexpression did not protect against apoptosis if MEK-1 expression was silenced. These results indicate that polyamines destabilize the MEK-1 mRNA and repress its translation by inhibiting the association between HuR and the MEK-1 transcript. Our findings indicate that MEK-1 is a key effector of the HuR elicited anti-apoptotic programme in IECs. PMID- 20001966 TI - Iron-binding activity in yeast frataxin entails a trade off with stability in the alpha1/beta1 acidic ridge region. AB - Frataxin is a highly conserved mitochondrial protein whose deficiency in humans results in Friedreich's ataxia (FRDA), an autosomal recessive disorder characterized by progressive ataxia and cardiomyopathy. Although its cellular function is still not fully clear, the fact that frataxin plays a crucial role in Fe-S assembly on the scaffold protein Isu is well accepted. In the present paper, we report the characterization of eight frataxin variants having alterations on two putative functional regions: the alpha1/beta1 acidic ridge and the conserved beta-sheet surface. We report that frataxin iron-binding capacity is quite robust: even when five of the most conserved residues from the putative iron binding region are altered, at least two iron atoms per monomer can be bound, although with decreased affinity. Furthermore, we conclude that the acidic ridge is designed to favour function over stability. The negative charges have a functional role, but at the same time significantly impair frataxin's stability. Removing five of those charges results in a thermal stabilization of approximately 24 degrees C and reduces the inherent conformational plasticity. Alterations on the conserved beta-sheet residues have only a modest impact on the protein stability, highlighting the functional importance of residues 122-124. PMID- 20001967 TI - Analysis of RGS2 expression and prognostic significance in stage II and III colorectal cancer. AB - The role of RGS2 (regulator of G-protein signalling 2) has been studied in several tumours. The purpose of the present study is to investigate the correlations between clinicopathological factors and patients' survival time and RGS2 expression in stage II and III CRC (colorectal cancer) patients. Real-time quantitative PCR was performed in 36 CRC tissues with recurrence and 28 without recurrence, and in three CRC-metastasis-derived cell lines (SW620, LoVo and Colo205) and 3 primary-CRC-derived ones (SW480, Caco-2 and HCT116) to examine RGS2 mRNA expression. In addition, to provide visualized evidence for RGS2 mRNA expression, random CRC samples were also performed with RT-PCR (reverse transcription-PCR). RGS2 protein was detected by immunostaining in 118 paraffin embedded specimens, and the correlations between clinicopathological factors and survival time and RGS2 expression were analysed. We found that RGS2 mRNA was down regulated both in CRC tissues with recurrence and metastasis-derived cell lines, and the expression level of RGS2 was unrelated to gender, age, tumour grade, or lymphovascular or perineural invasion. However, it was positively related to disease-free survival time (P<0.05). Furthermore, low RGS2 expression indicated a poorer survival rate (P<0.05, log-rank test). Multivariate analysis also showed that weak RGS2 protein expression was an independent adverse prognosticator in CRC (P<0.05). Taken together, we suggested that down-regulation of RGS2 might play an important role in CRC metastasis and predict poor prognosis in stage II and III CRC patients. PMID- 20001968 TI - Amyloid beta-peptide directly induces spontaneous calcium transients, delayed intercellular calcium waves and gliosis in rat cortical astrocytes. AB - The contribution of astrocytes to the pathophysiology of AD (Alzheimer's disease) and the molecular and signalling mechanisms that potentially underlie them are still very poorly understood. However, there is mounting evidence that calcium dysregulation in astrocytes may be playing a key role. Intercellular calcium waves in astrocyte networks in vitro can be mechanically induced after Abeta (amyloid beta-peptide) treatment, and spontaneously forming intercellular calcium waves have recently been shown in vivo in an APP (amyloid precursor protein)/PS1 (presenilin 1) Alzheimer's transgenic mouse model. However, spontaneous intercellular calcium transients and waves have not been observed in vitro in isolated astrocyte cultures in response to direct Abeta stimulation in the absence of potentially confounding signalling from other cell types. Here, we show that Abeta alone at relatively low concentrations is directly able to induce intracellular calcium transients and spontaneous intercellular calcium waves in isolated astrocytes in purified cultures, raising the possibility of a potential direct effect of Abeta exposure on astrocytes in vivo in the Alzheimer's brain. Waves did not occur immediately after Abeta treatment, but were delayed by many minutes before spontaneously forming, suggesting that intracellular signalling mechanisms required sufficient time to activate before intercellular effects at the network level become evident. Furthermore, the dynamics of intercellular calcium waves were heterogeneous, with distinct radial or longitudinal propagation orientations. Lastly, we also show that changes in the expression levels of the intermediate filament proteins GFAP (glial fibrillary acidic protein) and S100B are affected by Abeta-induced calcium changes differently, with GFAP being more dependent on calcium levels than S100B. PMID- 20001969 TI - Effects of GIK (glucose-insulin-potassium) on stress-induced myocardial ischaemia. AB - Despite the evidence in experimental animal models that insulin, or GIK (glucose insulin-potassium), improves left ventricular function and perfusion during both acute and chronic ischaemia, clinical studies have generated conflicting results. We tested the hypothesis that pretreatment with GIK attenuates the vascular and functional effects of stress-induced myocardial ischaemia in humans. Twenty-two patients with evidence of inducible myocardial ischaemia were enrolled; 11 patients with normal ventricular function underwent two dipyridamole echocardiography tests, and 11 with regional contractility defects from previous myocardial infarction were submitted to two ECG exercise tests combined with 201Tl myocardial perfusion scintigraphy; the tests were preceded by 60 min of either normal saline or an isoglycaemic GIK infusion. On a stress echocardiogram, a 30% reduction in the severity of ischaemia was observed. On ECG ergometry, GIK infusion slightly increased the time to ischaemia (+0.6 min, P=0.07); however, the higher workload (+8%, P=0.07) was achieved at a similar rate-pressure plateau. On scintigraphy, an increase in ischaemic segments (+48%, P<0.001) was imaged mainly at the expense of viable (but non-ischaemic) and non-viable segments, which were reduced by 60%. GIK affected stress-induced left ventricular underperfusion only marginally (GIK: 39.7+/-2.5 compared with saline: 35.4+/-2.2 units, P<0.05), but significantly improved its acute reversibility (-42+/-4 compared with -25+/-4%, P<0.001). We conclude that GIK pretreatment attenuates the effect of ischaemia on myocardial contractility, slightly improves exercise tolerance and causes a more rapid and diffuse recovery of post-ischaemic reperfusion. PMID- 20001971 TI - Atomic force microscopy comes of age. AB - AFM (atomic force microscopy) analysis, both of fixed cells, and live cells in physiological environments, is set to offer a step change in the research of cellular function. With the ability to map cell topography and morphology, provide structural details of surface proteins and their expression patterns and to detect pico-Newton force interactions, AFM represents an exciting addition to the arsenal of the cell biologist. With the explosion of new applications, and the advent of combined instrumentation such as AFM-confocal systems, the biological application of AFM has come of age. The use of AFM in the area of biomedical research has been proposed for some time, and is one where a significant impact could be made. Fixed cell analysis provides qualitative and quantitative subcellular and surface data capable of revealing new biomarkers in medical pathologies. Image height and contrast, surface roughness, fractal, volume and force analysis provide a platform for the multiparameter analysis of cell and protein functions. Here, we review the current status of AFM in the field and discuss the important contribution AFM is poised to make in the understanding of biological systems. PMID- 20001970 TI - Thr435 phosphorylation regulates RelA (p65) NF-kappaB subunit transactivation. AB - Phosphorylation of the RelA (p65) NF-kappaB (nuclear factor kappaB) subunit has been previously shown to modulate its ability to induce or repress transcription. In the present study we have investigated the consequences of Thr435 phosphorylation within the C-terminal transactivation domain of RelA. We confirm that Thr435 is phosphorylated in cells and is induced by TNFalpha (tumour necrosis factor alpha) treatment. Mutational analysis of this site revealed gene specific effects on transcription, with a T435D phosphomimetic mutant significantly enhancing Cxcl2 (CXC chemokine ligand 2) mRNA levels in reconstituted Rela-/- mouse embryonic fibroblasts. Chromatin immunoprecipitation analysis revealed that this mutation results in enhanced levels of histone acetylation associated with decreased recruitment of HDAC1 (histone deacetylase 1). Moreover, mutation of this site disrupted RelA interaction with HDAC1 in vitro. Thr435 phosphorylation of promoter-bound RelA was also detected at NF kappaB target genes following TNFalpha treatment in wild-type mouse embryonic fibroblasts. Phosphorylation at this site therefore provides an additional mechanism through which the specificity of NF-kappaB transcriptional activity can be modulated in cells. PMID- 20001973 TI - Genetic factors and epigenetic factors for autism: endoplasmic reticulum stress and impaired synaptic function. AB - The molecular pathogenesis of ASD (autism spectrum disorder), one of the heritable neurodevelopmental disorders, is not well understood, although over 15 autistic-susceptible gene loci have been extensively studied. A major issue is whether the proteins that these candidate genes encode are involved in general function and signal transduction. Several mutations in genes encoding synaptic adhesion molecules such as neuroligin, neurexin, CNTNAP (contactin-associated protein) and CADM1 (cell-adhesion molecule 1) found in ASD suggest that impaired synaptic function is the underlying pathogenesis. However, knockout mouse models of these mutations do not show all of the autism-related symptoms, suggesting that gain-of-function in addition to loss-of-function arising from these mutations may be associated with ASD pathogenesis. Another finding is that family members with a given mutation frequently do not manifest autistic symptoms, which possibly may be because of gender effects, dominance theory and environmental factors, including hormones and stress. Thus epigenetic factors complicate our understanding of the relationship between these mutated genes and ASD pathogenesis. We focus in the present review on findings that ER (endoplasmic reticulum) stress arising from these mutations causes a trafficking disorder of synaptic receptors, such as GABA (gamma-aminobutyric acid) B-receptors, and leads to their impaired synaptic function and signal transduction. In the present review we propose a hypothesis that ASD pathogenesis is linked not only to loss of-function but also to gain-of-function, with an ER stress response to unfolded proteins under the influence of epigenetic factors. PMID- 20001974 TI - Cyclo-oxygenase 2 up-regulates the effect of multidrug resistance. AB - COX-2 (cyclo-oxygenase 2), an inducible form of the enzyme that catalyses the first step in the synthesis of prostanoids, is associated with inflammatory diseases and carcinogenesis, which is suspected to promote angiogenesis and tissue invasion of tumours and resistance to apoptosis. COX-2 is also involved in drug resistance and poor prognosis of many neoplastic diseases or cancers. The activation of the COX-2/PGE2 (prostaglandin E2)/prostaglandin E receptor signal pathway can up-regulate the expression of all three ABC (ATP-binding-cassette) transporters, MDR1/P-gp (multidrug resistance/P-glycoprotein), MRP1 (multidrug resistance protein 1) and BCRP (breast-cancer-resistance protein), which encode efflux pumps, playing important roles in the development of multidrug resistance. In addition, COX inhibitors inhibit the expression of MDR1/P-gp, MRP1 and BCRP and enhance the cytotoxicity of anticancer drugs. Therefore we can use the COX inhibitors to potentialize the effects of chemotherapeutic agents and reverse multidrug resistance to facilitate the patient who may benefit from addition of COX inhibitors to standard cytotoxic therapy. PMID- 20001975 TI - [Risk factors of thrombotic complications and prognosis of patients with chronic form of ischemic heart disease]. PMID- 20001976 TI - [Value of telemetric support organization in prehospital management of patients with myocardial infarction for delays shortening of thrombolytic therapy]. AB - Aim of the study was to assess efficacy of the use of telemetric support of prehospital thrombolytic therapy (TLT) of myocardial infarction. We observed 206 patients with ST-elevation myocardial infarction (MI). These patients were divided into 2 groups: in one TLT was initiated after admission to hospital, in the other TLT was carried out by ambulance personnel with telemetric support. We analyzed times from onset of symptoms, rates of development of complications of MI and TLT. The use of telemetric support of ambulance personnel allowed to substantially shorten time from onset of symptoms to initiation of TLT, to minimize errors during its course and allowed to coordinate actions of physicians of ambulances and specialized hospital departments with comparable number of adverse events. PMID- 20001977 TI - [Efficacy of strict glycemic control in first 24 hours of myocardial infarction in patients with type 2 diabetes mellitus]. AB - The study evaluated the influence of strict glycemic control during first 24 hours of myocardial infarction on 6-months mortality and systolic function of the left ventricle. Of 65 patients with type 2 diabetes mellitus and acute myocardial infarction 30 received 24-hour infusion of short acting insulin and 35 served as controls. As a result of this investigation we made a conclusion that effect of optimal metabolic control during acute period of myocardial infarction on mortality and constriction of the left ventricular was beneficial. PMID- 20001978 TI - [24-hour rhythm of arterial pressure in patients with arterial hypertension associated with autoimmune thyroiditis]. PMID- 20001979 TI - [Provision of complex cardioprotection with the help of carvedilol in 1-2 degree arterial hypertension combined with metabolic syndrome]. AB - In an open prospective 16-week study we carried out assessment of clinical efficacy, vaso- and cardioprotective properties of nonselective alpha and beta adrenoblocker carvedilol used as monotherapy and in combination with nifedipine or as combination of these drugs with metformin and simvastatin. We have shown that the use of carvedilol both as monotherapy and in combination with nifedipine normalizes arterial pressure irrespective of the number of risk factors. We also revealed pronounced cardio- and vasoprotective action without negative effect on carbohydrate and lipid metabolism. PMID- 20001980 TI - [Sudden out of hospital cardiac death in children, adolescents, and subjects younger than 45 years]. AB - Aim of this study was determination of incidence and structure of sudden out of hospital cardiac death (SCD) in subjects aged 1-45 years in Moscow. We analyzed rate and structure of SD among persons who had succumbed in 2005-2007 in several districts of Moscow with population of 2,502,836. Of the total number of 19,557 autopsies 7702 (39.4% or 92/100 000/year) and 1265 (6.5%, 19.4% of autopsies in this age group, or 16.8/100000/year) in all age groups and in the age 1-45 years, respectively, were performed because of SCD. In most cases (44%) age at SCD was 41-45 years, only in 1% of cases it was less than 18 years. Eighty two percent of autopsied SD victims were men. Most frequent postmortem diagnosis was "cardiomyopathy" (69% overall, 80-96% in the age 19-35 years) established on the basis of detection of both specific and nonspecific changes in the myocardium. In 25% of SD cases in the age group before 18 years congenital heart disease was found. After 35 years rose portion of diseases of ischemic nature (22-32%). Percentages of hypertensive disease and other diseases among all diagnoses were 7 and 1, respectively. Beginning with 19 years in large percentage of cases presence of alcohol in blood was detected. This percentage was especially high in the age group 19-25 years (66.6%). SCD of most persons aged 20-45 years was registered at home, while in 50% of younger persons it occurred outside home often during physical effort. Rate of SCD, its circumstances and structure elicited in this study might serve as basis for elaboration of the system of early detection of groups of risk and prevention of SD in young age. PMID- 20001981 TI - [Dietary SkQ1 supplement reduces myocardial ischemia- reperfusion injury in rats in vivo]. AB - To examine whether nutritional supplementation with SkQ1 can reduce myocardial ischemia-reperfusion injury in vivo, Wistar rats were fed a regular diet supplemented with different doses of SkQ1 for two or three weeks. Control groups of rats were fed the same diet supplemented with NaBr. Anaesthetized rats were subjected to 40-min regional myocardial ischemia and 1-h reperfusion. Myocardial infarct size was measured by 2,3,5-triphenyl tetrazolium chloride (TTC) staining method. SkQ1-fed rats (125 nmol/kg/day for two weeks and 250 nmol/kg/day for two and three weeks) revealed significantly smaller myocardial infarction and less lactate dehydrogenase (LDH) and creatine kinase-MB fraction (CK-MB) activity elevations in plasma at the end of reperfusion compared with the controls. This effect was combined with improvement of energy state of the area at risk at the end of reperfusion, namely, augmentation of adenine nucleotide content, two-fold increase in phosphocreatine, reduction of lactate accumulation and decrease of lactate/pyruvate ratio in myocardial tissue. Therefore, nutritional supplementation with SkQ1 renders the hearts resistant to ischemia-reperfusion injury affecting oxidative metabolism of postischemic cardiomyocytes. PMID- 20001982 TI - [The role of T-wave alternans in prognosis of risk of sudden cardiac death]. AB - The literature review contains data dedicated to one of tools of stratification of risk of fatal arrhythmia - detection of T-wave alternans (TWA). Methods of determination of TWA, problems of results interpretation, mechanism of the TWA genesis, and its clinical meaning in different cardiovascular diseases are considered. Special attention is paid to the role of TWA in the identification of the patients who require cardioverter-defibrillator implantation. PMID- 20001983 TI - [Prevention and correction of cognitive disorders in arterial hypertension]. AB - In this review we systematized literature data illuminating problems of prevention and correction of cognitive disorders in patients with arterial hypertension (AH). On the basis of results of multicenter randomized studies we have demonstrated close interrelationship between cardiac and cerebral pathology and value of treatment of concomitant neurologic and cardiovascular disorders for prevention of cognitive disturbances in patients with AH. We have shown rational approach to planning antihypertensive therapy and to degree of arterial pressure (AP) lowering in patients with AH, including those with cerebrovascular pathology having high risk of development of stroke and dementia. It has been also shown that in patients with AH calcium antagonists, angiotensin II receptor blockers, and inhibitors of angiotensin converting enzyme posses definite advantages over antihypertensive drugs from other classes in prevention of dementia and less severe cognitive disturbances. PMID- 20001984 TI - [Microangiopathy in type 2 diabetes mellitus: mechanisms of development, possibilities of prevention, treatment]. PMID- 20001985 TI - [Nitrous oxide and atherosclerosis. Nitrous oxide, dysfunction of vascular endothelium, and pathogenesis of atherosclerosis]. PMID- 20001986 TI - [The syndrome of mitral regurgitation in patients with chronic heart failure]. AB - Functional mitral regurgitation is a frequent complication of severe course of ischemic heart disease and plays a role of no small importance in origin and progression of chronic heart failure (CHF). Severe degree of mitral regurgitation is found in 18.9%, moderate - in 29.7% of patients with systolic left ventricular dysfunction. A major precondition for origin of mitral regurgitation is the presence of the process of left ventricular remodeling which disturbs coordination between the ventricle and valvular apparatus. Changes of anatomo functional state of the mitral valve occurring during its closure determine severity of mitral regurgitation in patients with CHF and left ventricular dysfunction. PMID- 20001987 TI - [Immediate and remote results after percutaneous transluminal coronary angioplasty and coronary bypass surgery in patients with multivascular involvement of coronary arteries]. AB - Relatively little time has passed since elaboration and introduction of coronary angioplasty however interventional cardiology in these years has made a swift leap in its development. Initially coronary interventions were carried out only on isolated proximal stenoses but later interventions became to be performed on more complex lesions of coronary arteries and then on multivessel disease intruding into area which previously had belonged exclusively to coronary bypass surgery. Although both methods of revascularization rapidly developed the researchers were interested in assessment and comparison of results of two strategies. Thus studies appeared in which patients were randomized for coronary bypass surgery or percutaneous coronary angioplasty. Below we present data of the largest and most interesting studies in which these strategies of revascularization in patients with multivessel involvement have been compared. We also separately consider results of revascularization in a subgroup of patients with diabetes mellitus. PMID- 20001988 TI - [Early reperfusion and aborted myocardial infarction]. AB - Data on effect of early reperfusion on prognosis of patients with myocardial infarction, possibilities of development of "aborted myocardial infarction", data on prognosis of "aborted myocardial infarction" and its prevalence are presented. PMID- 20001989 TI - [Effects of non-drug therapy of moderate hypercholesterolemia with the use of phytosterins]. PMID- 20001990 TI - Comparison of complications and dialysis adequacy between temporary and permanent tunnelled catheter for haemodialysis. AB - Number of hemodialysis patients each day is increasing. The quality of their lives is largely determined by the quality of hemodialysis treatment. One of the most important factors is the type of applied blood approach. The type of blood approach in the most case is artery venous fistula, permanent, temporary catheters, grafts. Any complications of blood strand approach inevitably leads to lower quality of hemodialysis treatment which is connected with not adequate dialysis and poorer general state of patients. Our research was carried out as a prospective study, for the period of 36 months. In the study were included 31 patients, which are on chronic haemodialysis treatment. During this study, we are followed all complications, which occurred at temporary, and permanent tunneled haemodialysis catheters. Complications have occurred in terms of thrombotic problems, low blood flow, occurrence of infection. All patients are divided in two groups, 16 patients with permanent and 15 patients with temporary catheters. In the course of the study was analyzed blood flow and dialysis adequacy (Kt/Vdp) as well as complications and results was compared with randomly selected 16 patients who haemodialysis treatment performed by artery venous fistula (AVF). Two patients were lost to further follow-up to the end of the study. 26 patients at the end of the study had functional catheters, while in the case of 3 patients the catheter was removed. Infection was found in 10 patients while thrombotic complications were observed in 27 cases regardless of catheter type. Mean blood flow in patients with permanent catheter was significantly higher (296,9+/-28,45 cm3/min) compared to patients with temporary catheter (226,3+/-39,8 cm3/min) (p<0,001). Kt/Vdp delivered was 1,22+/-0,15 on patients with permanent catheter and 1,30+/-0,18 for artery venous fistula (AVF) access respectively. The loss of dialysis efficacy using catheters was estimated at 6%. However, in all cases Kt/Vdp values remained above the recommended values (Kt/Vdp > or = 1,2). PMID- 20001991 TI - Transfusion-transmitted infections in haemophilia patients. AB - One of the largest therapeutic problem during the continuous treatment of the patients with Hemophilia A and B, are viral infections as Hepatitis B and C, and HIV, and the other infective diseases, which can be transmitted by the transfusion of blood products. The aim of this study is to analyze the complications of the hemophiliacs in Kosovo which have been treated with fresh frozen plasma, cryoprecipitate and concentrated products of FVIII and FIX. We have tested 75 patients with hemophilia A or B and there were used enzyme immunoassay test-Elisa method for the following: anti-HCV, HBsAg, HIV and TPHA.The serological data showed that HCV infection was positive in 29 cases or 38,7%, whereas infection with HBV and HIV were present in a smaller percentage of the patients (2,7% HBV and 1,4% for HIV). HCV infection was present only in 9,5% of the cases of the age group under 18 years. Infected hemophiliacs with one or two infective agents were found in 34,7%, respectively 4%. Infection with T. pallidum was present at none of the examined patients with hemophilia. HCV infection was higher in severe forms of hemophilia B (44,4%), compared with severe form of hemophilia A (30%).Based on our results, despite the infrequent application of FVIII and FIX concentrates, and other anti hemophilic preparations used in treating hemophilia patients, the number of infected hemophiliacs with blood-transmittable infectious agents was substantially high, especially with hepatitis C virus. PMID- 20001992 TI - Hepatitis C virus genotypes in chronic hepatitis C patients and in first time blood donors in northeastern Bosnia and Herzegovina. AB - Prevalence of hepatitis C virus (HCV) genotypes in Bosnia and Herzegovina (B&H) is an issue that is not sufficiently researched and there is a need for studies that would explore this in detail.The aim of this study was to determine the distribution of HCV genotypes in the group of patients with chronic hepatitis C and also in the group of first time blood donors that tested positive for anti HCV antibodies during the blood screening process. Our secondary goal was to compare the proportions of HCV genotypes between these two groups.We analyzed 75 blood samples of patients with confirmed chronic hepatitis C. We also analyzed 13/16082 blood samples of first time blood donors found to be HCV positive during the blood screening process. We also determined HCV genotype in HCV RNA positive samples.We have found that genotype 1b was more prevalent in chronic hepatitis C patients (52/75; 69,3%) than in first time blood donors (6/13; 46,1%), however this difference was not statistically significant (c2=1,721; df=1; p=0,19). Genotype 1a was more prevalent in the group of first time blood donors (3/13; 23,1%) than in the group of chronic hepatitis C patients (3/75; 4%), but this was also with limited statistical significance (c2=3,71; df=1; p=0,054). We have not found any significant difference in prevalence of genotypes 1a (p=0,2) and genotypes 3 (p=0,70) when compared between chronic patients (3/75 and 16/75; respectively) and first time blood donors (3/13 and 4/13; respectively). Our study confirmed domination of genotype 1b in the region of northeastern B&H which is in accordance with HCV genotype prevalence in other countries in our part of Europe. PMID- 20001993 TI - Family analysis of immunoglobulin classes and subclasses in children with autistic disorder. AB - Autistic disorder is a severe neurodevelopment disorder characterized by a triad of impairments in reciprocal social interaction, verbal and nonverbal communication, and a pattern of repetitive stereotyped activities, behaviours and interests. There are strong lines of evidence to suggest that the immune system plays an important role in the pathogenesis of autistic disorder. The aim of this study was to analyze quantitative plasma concentration of immunoglobulin classes, and subclasses in autistic patients and their families. The investigation was performed retrospectively in 50 persons with autistic disorder in the Republic of Macedonia. Infantile autistic disorder was diagnosed by DSM-IV and ICD-10 criteria. Plasma immunoglobulin classes (IgM, IgA, and IgG) and subclasses (IgG1, IgG2, IgG3, and IgG4) were determined using Nephelometer Analyzer BN-100. Multiple comparisons for the IgA variable have shown statistically significant differences between three pairs: male autistic from the fathers (p = 0,001), female autistic from the mothers (p = 0,008), as well as healthy sisters from the fathers (p = 0,011). Statistically significant differences found between three groups regarding autistic disorder (person with autistic disorder, father/mother of a person with autistic disorder, and brother/sister) independent of sex belongs to IgA, IgG2, and IgG3 variables. Multiple comparisons for the IgA variable have shown statistically significant differences between children with autistic disorder from the fathers and mothers (p < 0,001), and healthy brothers and sisters from the fathers and mothers (p < 0,001). Comparison between healthy children and children with autistic disorder from the same family should be tested for immunoglobulin classes and subclasses in order to avoid differences between generations. PMID- 20001994 TI - Biochemical evaluation of the therapeutic effectiveness of honey in oral mucosal ulcers. AB - The aim of this study was to describe and quantify the therapeutic value of honey in oral mucosal ulcers healing in comparison with Glyceroloxytriester (TGO). We also aimed to biochemically evaluate the healing effects of honey which had been collected from the Blacksea region flora on mucosal ulcers resulting in material loss.Thirty wistar rats (240+/-30 g) were enrolled in this study. Excisional wounds were performed in all rats for animal oral mucosal ulcer model. They were randomly allocated to three groups: group 1 was treated with Apitherapeutic agent or honey (0,1 ml, 2x1), group 2 was treated with TGO (0,1ml, 2x1) locally, Group 3 served as the control group.Following the surgical procedure on day 7, biopsy specimens were taken from right buccal mucosa and on day 14 biopsy specimens were taken from left bucal mucosa in all rats. Afterwards, hydroxy pyroline levels were measured. Data were analyzed statistically.There was no statistically significant difference between Group 1 and 2, and also between Group 2 and 3, but there was statistically significant difference between Group 1 and 3 on day 7. There was no statistically significant difference between Group 1, 2 and 3 on day 14. PMID- 20001995 TI - Pulpal blood flow changes in abutment teeth of removable partial dentures. AB - The purpose of this study was to investigate the effect of tooth supported (TSD) and tooth-tissue supported (TTSD) removable partial denture wearing on pulpal blood flow (PBF) of the abutment teeth by using Laser Doppler Flowmeter (LDF). Measurements were carried out on 60 teeth of 28 patients (28 teeth and 12 patients of TTSD group, 32 teeth and 16 patients of TSD group) who had not worn any type of removable partial dentures before, had no systemic problems and were non smokers. PBF values were recorded by LDF before insertion (day 0) and after insertion of dentures at day 1, day 7 and day 30. Statistical analysis was performed by student t test and covariance analyses of repeated measurements. In the group TTSD, the mean values of PBF decreased statistically significantly at day 1 after insertion when compared with PBF values before insertion (p<0,01). There was no statistically significant difference among PBF mean values on 1st, 7th and 30th day. However, in the group TSD, there was no statistically significant difference among PBF mean values before insertion and on 1st, 7th and 30th day. In other words, PBF mean values in group TSD continued without changing statistically significant on 1st, 7th and 30th day.TTSD wearing may show negative effect on the abutment teeth due to decreasing basal PBF. PMID- 20001996 TI - Correlation between interleukin 6 and interleukin 10 in acute myocardial infarction. AB - The aim of this study was to analyze (i) ratios between pro-inflammatory cytokines interleukin 6 (IL-6), interleukin 1 (IL-1), tumour necrosis factor alpha (TNF-alpha) and anti-inflammatory cytokine interleukin 10 (IL-10) in patients with acute myocardial infarction (AMI) and stable angina pectoris (ii) as well as correlation between IL-6 and IL-10 in AMI and (iii) correlation between IL-6 and lipoproteins in AMI.The total of 71 patients were enrolled in this study, 41 of them with AMI (study group) and 30 with stable angina pectoris (control group). The concentrations of cytokines and lipoproteins were measured from blood samples. Pro-inflammatory to anti-inflammatory cytokine ratios were calculated by dividing concentrations of pro-inflammatory cytokines with IL-10. In statistical analyses we used descriptive statistics, normality tests and analysis of correlation.IL-6: IL-10 ratio is significantly higher in AMI than in stable angina (P < 0,001), TNF-alpha: IL-10 is also higher in study group but the difference is not significant. We found positive linear correlation between IL-6 and IL-10 (r =0,43; p = 0,015) and negative linear correlation between IL-6 and high density lipoprotein HDL (r = -0,47; p= 0,008) in AMI.IL-6: IL-10 ratio is higher in AMI than in stable angina. There is linear correlation between IL-6 and IL-10 and IL-6 and HDL in AMI. PMID- 20001997 TI - The possible role of early post-transplant inflammation in later anemia in kidney transplant recipients. AB - Delayed kidney graft function and acute rejection in the early post-transplant period affect both short and long-term allograft survival. Allograft rejection, as an inflammatory state, results in increased erythropoietin resistance, which leads to decreased haemoglobin (Hb) level. We conducted this study to evaluate whether inflammation in the early post-transplant period could predict later anemia.This is a retrospective cohort study based on the analysis of 64 existing clinical records. PREDICTOR: White blood cells (WBC) count obtained by the end of the first week post-transplant (W1). Covariates: Donor's age, recipient's age and sex. OUTCOME: Anemia identified at 12 months (M12) post engraftment. Median WBC count at W1 was 9,5 x103/microL (5th - 95th percentile 5,2 x103/microL -17,8 x103/microL). Mean Hb values at M12 were 129,9 +/- 20,3 g/L, in males 136,2 +/- 20,1 g/L and in females 119,4 +/- 16,2 g/L. The significant correlation was found between WBC at W1 and Hb at M12. Pearson coefficient of correlation r was -0,26, and 95% confidence interval (CI) for r was -0,47 to -0,015 (p=0,03). Univariate logistic regression showed significant association between WBC at W1 and Hb at M12 (OR 1,20; 95% CI 1,04 to 1,39, p=0,01). After the adjustment for donor's and recipient's age by transplantation and recipient's sex, multiple regression showed that WBC count remained predictive of anemia at M12 (OR 1,17; 95% CI 1,01 to 1,36, p=0,03). Early post-transplant inflammatory response predicts later anemia in kidney transplant recipients. An increase in WBC count in the first week post-transplant by 109/L increases the risk for anemia after twelve months by 17%. PMID- 20001998 TI - Outpatient management of oral anticoagulation therapy in patients with nonvalvular atrial fibrillation. AB - Due to heightened risk for thromboembolic complications, nonvalvular atrial fibrillation (NVAF) presents an absolute indication for long-term oral anticoagulation therapy. This was an observational, analytical, randomised, one year clinical study, conducted in the Blood Transfusion Institute Sarajevo, Bosnia & Herzegovina. The aim of this study was to present the oral anticoagulation treatment in terms of International normalised ratio (INR) monitoring and warfarin/acenocoumarol dose titration in 117 patients with NVAF. INR values, the doses of warfarin and acenocoumarol, as well as the tendency and adequacy of their changes were monitored. Percentages of the therapeutic INR values were 51,77% and 53,62%, subtherapeutic 42,84% and 35,86%, and supratherapeutic 5,39% and 10,53% for the warfarin and acenocoumarol treatment, respectively. The average total weekly doses (TWD) which most frequently achieved the therapeutic INR values were 27,89+/-12,34 mg and 20,44+/-9,94 mg, for warfarin and acenocoumarol, respectively. The dose changes with the INR values 1,7 or lower/3,3 or higher were omitted in 13,46% and 15,63%, and with the INR values 1,8-3,2 were noted in 8,62% and 13,48% of all the check-up visits in the warfarin and acenocoumarol group, respectively. The annual dose changes were noted in 24,65% and 31,41%, and the daily dose changes in 74,43% and 73,36% of all the check-up visits of warfarin and acenocoumarol group, respectively. We can conclude that the management of the oral anticoagulation treatment in our country is in accordance with the relevant recommendations, but with the present tendency toward underdosing and unnecessary frequent dose changing. PMID- 20001999 TI - Evaluation of risk markers fluctuation during an initial therapy with rosiglitazon in patients suffering from metabolic syndrome. AB - The aim of this study was to examine the effects of hypoglycaemic drug-agonists of PPAR-gama receptors-rosiglitazone (Avandia,4 mg - Glaxo Smith Kline) on values of wide-spread risk - markers-fibrinogen, C-reactive protein and uric acid and glicolysated haemoglobin HbA1C as parameter of metabolic control .We included fourty patients with criteria for metabolic syndrome and evaluated results into groups of diabetic and prediabetic patients according to criteria of IDF (International Diabetic Federation)These risk markers and glicolysated haemoglobin HbA1C were observed at the start of therapy, then after four, eight and twelve weeks and results were compared and statistically calculated. Three months initial therapy with rosiglitazone significantly reduced values of HbA1C, fibrinogen and CRP but not uric acid in prediabetic patients.Rosiglitazone initial three months therapy significantly reduced HbA1C, fibrinogen and uric acid, but not CRP in diabetic patients. PMID- 20002000 TI - Prevalence of resistence to activated protein C (APC-resistance) in blood donors in Kosovo. AB - One of the most frequent hereditary causes of thrombophilia is, without a doubt, resistance to Activated Protein C (APC-resistance), which is a consequence of point mutation in gene coding for coagulation Factor V (Factor V Leiden) in 90 95% of cases. The aim of this paper was to determine prevalence of APC-resistance in a group of healthy blood donors. The size of the group is quite representative of Kosovo Albanians. A total of 944 blood donors were examined (537 males and 407 females), for whom APC-resistance was determined by functional methods of coagulation using the kit ACTICLOT(R) Protein C Resistance. Method is based on the test of APTT determined twice: first in the presence and second in the absence of activated Protein C (APC). The ratio of these two values constitutes is called Activated Protein C- Sensitivity Ratio (APC-SR). From 944 examined donors, pathologic values of APC-SR (1,3-1,9) were found in 32 persons (3,4% of the total number). The distribution among sexes was 3,35% (18/537) in male and 3,43% (14/407) in female subjects. The mean values of APC-SR (1,64 in male and 1,71 in female subjects) were not significantly different (P = 0,22). Based on these results, we conclude that the prevalence of APC resistance in Albanian population of Kosovo is within the lower limit of prevalence in general population in different countries of European countries, which, according to some authors ranges is from 3 to 7%. PMID- 20002001 TI - The study of volume density of tracheal ganglions in vitro in new born babies with respiratory distress syndrome. AB - Volume density of respiratory organs was studied in vitro in newborn babies at different age of gestation (abort, immature, premature and mature) using stereometric method. The total of 23 cases was subject to this study. The respiratory organs (trachea, lungs) were taken from autopsies of newborn babies exited from different causes. For this purpose the tissues were fixed in formalin (10%) solution, cut serially in 7micro and 10micro slabs. Volume density of the respiratory system was assessed stereometricaly using Universal testing system Weibel M 42. We observed that volume density of epithelia, musculature and glands were proportionally present in the tracheal tissue. Cellular interstitial tissue is consistently increasing and corresponds to the developmental stages of the newborn babies. The density of tracheal ganglions is greater in premature ages of immature and premature newborns (p<0,05). Decreased number of ganglion cells is observed in mature ages (p<0,05). This is caused by intensive ramification of ganglions from serosa to deeper layers of trachea right to epithelium. Medium diameter of tracheal ganglions is greater in mature newborn babies and corresponds to developmental ages of babies. PMID- 20002002 TI - Role of meconium in the reaction of airways smooth musculature in the newborn with meconium aspiration syndrome (MAS). AB - The role of meconium in the respiratory system was studied in newborns, who died from various causes (250 up to 3000 g of weight). We monitored tracheal rings response to dopamine, serotonin and ethanol in different concentrations (dopamine: 0,05 mg/ml, 0,5 mg/ml, 5 mg/ml; serotonin (5-HT): 10-4, 10-3, 10-2, 10 1 mol/dm3; ethanol: 0,02 ml, 0,5 ml, 1,0 ml; 96%). Tracheal smooth musculature tonus (TSM) was examined in 48 tracheal preparations taken after the newborn exitus due to different reasons. Based on functional researche of isolated preparations of tracheas, it may be concluded that: aspiration of meconium has not changed the response of TSM to dopamine, serotonin and ethanol (p>0,1) in comparison with the control group, which have died due to different lung inflammatory processes (e.g. pneumonia, bronchopneumonia, atelectasis, cerebral hemorrhage). The results suggest that meconium does not potentiate the constricting action of dopamine, serotonin and ethanol in tracheobronchial system. Meconium causes mild relaxation of the TSM through a mechanism that is not intermediated by the products of cyclooxygenases (prostaglandins, prostacyclins) from the tracheal epithelium or proteins. Also, as it seems, the direct activity of many tested acids in the smooth musculature has no significant impact on increase of the airways tonus in MAS syndrome. PMID- 20002004 TI - Subclinical pulmonary dysfunction in spinocerebellar ataxias 1, 2 and 3. AB - OBJECTIVES: Evaluation of pulmonary function in patients with spinocerebellar ataxias (SCA) 1, 2 and 3 without clinical evidence of pulmonary dysfunction. METHODS: Thirty patients (F:M = 7:23; age: 35 +/- 11.3 years; SCA1 - 13, SCA2 - 9 and SCA3 - 8) without clinical manifestations of respiratory dysfunction and 30 controls underwent pulmonary function tests. The percentage predicted values of forced vital capacity (FVC), volume of air exhaled during first second of FVC (FEV1), peak expiratory flow rate (PEFR) and maximal voluntary ventilation (MVV), actual values of maximal inspiratory and expiratory pressures (MIP and MEP in mmHg), and ratios of actual values of FEV1/FVC (%) and FEV1/PEFR (ml/l/min) were analyzed. RESULTS: Compared with controls SCA patients had significant reductions of FVC (71.1 +/- 17.5 vs 85.5 +/- 18.7; P < 0.01), PEFR (51.5 +/- 20.7 vs 77.1 +/ 24.9; P < 0.001), MVV (64.4 +/- 21.6 vs 97.2 +/- 22.7; P < 0.001), MIP (27.7 +/- 16.8 vs 50.1 +/- 15.1; P < 0.001) and MEP (38.1 +/- 18.7 vs 74.7 +/- 16.0; P < 0.001), elevation of FEV1/PEFR (10.5 +/- 2.8 vs 7.4 +/- 2.1; P < 0.001), but no significant change of FEV1 and FEV1/FVC. FEV1/PEFR correlated positively with illness duration and MVV negatively with severity of illness. CONCLUSIONS: The present study showed subclinical restrictive type of pulmonary dysfunction in SCA, and possible presence of upper airway obstruction. Chest physiotherapy and breathing exercises should be introduced early in management of SCA. PMID- 20002005 TI - A review on sex differences in stroke treatment and outcome. AB - BACKGROUND: Beyond epidemiological differences, it has been controversial whether any important sex differences exist in the treatment of stroke. In this review paper, the following areas are covered: thrombolysis, stroke unit care, secondary prevention, surgical treatment, and rehabilitation. Additionally, symptoms at stroke onset, as well as outcome measures, such as death, dependency, stroke recurrence, quality of life, and depression are reviewed. METHODS: Search in PubMed, tables-of-contents, review articles, and reference lists after studies that include information about sex differences in stroke care. RESULTS: Ninety papers are included in this review. Women suffer more from cortical and non traditional symptoms. Men and women benefit equally from thrombolysis and stroke unit care. Women with cardioembolic strokes may benefit more from anticoagulant therapy. Most studies have not found any tendency towards sexism in the choice of treatment. Post-stroke depression and low quality-of-life seem to be more common among women. Mortality rates are higher among men in some studies, while long term ADL-dependency seems to be more common among women. CONCLUSIONS: Sex differences in stroke treatment and outcome are small, with no unequivocal proof of sex discrimination. Women have less favourable functional outcome because of higher age at stroke onset and more severe strokes. PMID- 20002006 TI - High prevalence of neuropathies in patients with end-stage liver disease. AB - OBJECTIVES: Peripheral neuropathy has been reported in association with end-stage liver disease, but there is only a limited number of reports on the incidence and features of these neuropathies. MATERIALS AND METHODS: In this study, 83 patients awaiting liver transplantation were evaluated for the presence of peripheral and autonomic neuropathy. RESULTS: Sixty-five percent of the patients had evidence of neuropathy, in agreement with peripheral NCS or cardiovascular autonomic function test. The neuropathy was more frequent in patients with advanced hepatic failure, evaluated with the MELD score. The most frequent abnormalities in nerve conduction studies were sensory-motor neuropathies and sensory neuropathies, with a length-dependent pattern. CONCLUSION: Peripheral neuropathy and autonomic neuropathy are common in patients with end-stage liver disease with different etiology and correlate with the severity of the liver disease. PMID- 20002007 TI - Cerebrospinal fluid metabolite and nigrostriatal dopaminergic function in Parkinson's disease. AB - OBJECTIVES: To evaluate the association between cerebrospinal fluid (CSF) homovanillic acid (HVA) concentrations and nigrostriatal dopaminergic function assessed by positron emission tomography (PET) imaging with carbon-11-labeled 2beta-carbomethoxy-3beta-(4-fluorophenyl)-tropane ((11)C-CFT), which can measure the dopamine transporter (DAT) density, in Parkinson's disease (PD). METHODS: (11)C-CFT PET scans and CSF examinations were performed on 21 patients with PD, and six patients with non-parkinsonian syndromes (NPS) as a control group. RESULTS: In the PD group, CSF HVA concentrations were significantly correlated with the striatal uptake of (11)C-CFT (r = 0.76, P < 0.01). However, in the NPS group, two indices were within the normal range. CONCLUSIONS: In PD, CSF HVA concentrations correlate with nigrostriatal dopaminergic function. Therefore, CSF HVA concentrations may be an additional surrogate marker for estimating the remaining nigrostriatal dopaminergic function in case that DAT imaging is unavailable. PMID- 20002008 TI - Anti-cyclic citrullinated peptide antibody (anti-CCP antibody) is present in the sera of patients with dementia of Alzheimer's type in Asian. AB - BACKGROUND: In the hippocampi of Alzheimer's disease (AD) patients, aberrant expression of citrullinated proteins and peptidylarginase 2 (PADI2) has been identified. We explored the functional roles of these proteins by means of detection of serum anti-cyclic citrullinated peptide antibody (anti-CCP antibody) in patients with dementia of Alzheimer's type (DAT). METHODS: Sera were obtained from 42 patients with DAT, 30 patients with other neurological disorders and 42 healthy controls. Gender ratio and age were comparable among the three groups. The level of anti-CCP antibody in sera was examined by ELISA. FINDINGS: Anti-CCP antibody was not found in the 30 patients with other neurological disorders, and only one of the 42 healthy controls (2.4%) was positive. However, surprisingly, anti-CCP antibody was clearly detected in eight of the 42 DAT patients. INTERPRETATION: Anti-CCP antibody appears to be a simple and early serologic biomarker for DAT among dementia patients. Additionally, our data imply that citrullinated proteins accumulated in the astrocytes of AD patients acquire neo antigenicity, inducing anti-CCP antibody production. PMID- 20002010 TI - Routine developmental screening at 5.5 and 7 years of age is not an efficient predictor of attention deficit hyperactivity disorder - a critical comment. PMID- 20002013 TI - Slow versus rapid enteral feeding advancement in preterm newborn infants 1000 1499 g: a randomized controlled trial. AB - AIM: To evaluate whether preterm neonates weighing 1000-1499 g at birth receiving rapid enteral feeding advancement at 30 mL/kg/day attain full feedings (180 mL/kg/day) earlier than those receiving slow enteral feeding advancement at 20 mL/kg/day without increase in the incidence of feeding intolerance or necrotizing enterocolitis. METHODS: A total of 100 stable intramural neonates weighing between 1000 and 1499 g and gestational age less than 34 weeks were randomly allocated to enteral feeding (expressed human milk or formula) advancement of 20 mL/kg/day (n = 50) or 30 mL/kg/day (n = 50). RESULTS: Neonates in the rapid feeding advancement group achieved full volume feedings before the slow advancement group (median 7 days vs. 9 days) (p < 0.001), had significantly fewer days of intravenous fluids (median 2 days vs. 3.4 days) (p < 0.001), shorter length of stay in hospital (median 9.5 days vs. 11 days) (p = 0.003) and regained birth weight earlier (median 16 days vs. 22 days) (p < 0.001). There were no statistical differences in the proportion of infants with apnea, feed interruption or feed intolerance. CONCLUSION: Rapid enteral feeding advancements of 30 mL/kg/day are well tolerated by stable preterm neonates weighing 1000-1499 g. PMID- 20002014 TI - Late and ultra late onset Streptococcus B meningitis: clinical and bacteriological data over 6 years in France. AB - BACKGROUND: Group B Streptococcus (GBS) is one of the leading causes of sepsis and meningitis in newborn. The objective of this study was to describe the characteristics of GBS meningitis in children aged between 7 and 89 days (late onset disease - LOD group) and to compare them with children aged more than 3 months (ultra late onset disease - ULOD group). METHODS: Clinical and biological data were gathered by ACTIV/GPIP (a nationwide active surveillance network). The study population included 242 children hospitalized between 2001 and 2006 for GBS meningitis (220 in the LOD group and 22 in the ULOD group). RESULTS: Univariate analysis revealed that gestational age (GA) was significantly lower in the ULOD group as compared with the LOD group (respectively 35.6 weeks vs. 37.9 weeks, p = 0.002). Prevalence of early preterm birth (before the 32nd week GA) was significantly higher in the ULOD group than in the LOD group (32% vs. 7%, p = 0.002). No significant difference was found between the two groups for biological characteristics of lumbar puncture, GBS serotypes, complications and survival rate. CONCLUSION: These data suggest that LOD and ULOD would be the same clinical and bacteriological entity, except for prematurity, which seems significantly associated with ULOD. PMID- 20002017 TI - 200 years of success initiated by James Wardrop's 1809 monograph on retinoblastoma. PMID- 20002018 TI - Antagonists of ionotropic gamma-aminobutyric acid receptors impair the NiCl2 mediated stimulation of the electroretinogram b-wave amplitude from the isolated superfused vertebrate retina. AB - PURPOSE: NiCl(2) (15 microM) stimulates the electroretinogram (ERG) b-wave amplitude of vertebrate retina up to 1.5-fold through its blocking of E/R-type voltage-gated Ca(2+) channels. Assuming that such an increase is mediated by blocking the release of the inhibitory neurotransmitter gamma-aminobutyric acid (GABA) via ionotropic GABA receptors, we tested the effect of both GABA itself and GABA-receptor antagonists such as (-)bicuculline (1.51-fold increase) and (1,2,5,6-tetrahydropyridin-4-yl)methylphosphinic acid (TPMPA; 1.46-fold increase) on the b-wave amplitude. METHODS: Recording of the transretinal potentials from the isolated bovine retina. RESULTS: GABA (100 microM) reduced the b-wave amplitude only when NiCl(2) (15 microM) was applied first. Each antagonist applied on its own stimulated the b-wave amplitude only partially: subsequent NiCl(2) superfusion caused a small but additional increase, leading to a 1.69- and a 1.88-fold total increase of the amplitude by Ni(2+) plus (-)bicuculline or Ni(2+) plus TPMPA, respectively. Only the application of both antagonists in combination, before superfusing low NiCl(2) (15 microM), completely prevented subsequent stimulation by NiCl(2) with a similar 1.90-fold total increase of b wave amplitude. Those retina segments that did not respond to NiCl(2) could not be stimulated by (-)bicuculline and vice versa. CONCLUSION: The stimulatory effect of NiCl(2) on the ERG b-wave amplitude is mainly, but not only, mediated by a NiCl(2)-sensitive, Ca(v)2.3-triggered GABA release acting through ionotropic GABA-A and GABA-C receptors. PMID- 20002019 TI - Commentary on Nelson et al. (2010). PMID- 20002020 TI - Interaction of SLC6A4 and DRD2 polymorphisms is associated with a history of delirium tremens. AB - Several genetic polymorphisms have been reported to be associated with alcohol withdrawal seizures (AWS) and delirium tremens (DT). To replicate and further explore these findings, we investigated the effects of 12 previously reported candidate genetic variations in two groups of alcohol-dependent European Americans with a history of withdrawal, which differed according to the presence (n = 112) or absence (n = 92) of AWS and/or DT. Associations of AWS and/or DT with the genomic and clinical characteristics and gene-gene interaction effects were investigated using logistic regression models. None of the polymorphisms were significantly associated with AWS/DT after correction for multiple testing. However, we found a significant interaction effect of the SLC6A4 promoter polymorphism (5-HTTLPR) and DRD2 exon 8 single nucleotide polymorphism rs6276 on AWS and/or DT history (P = 0.009), which became more significant after adjustment for lifetime maximum number of drinks consumed per 24 hours (P < 0.001). Subsequent analysis revealed an even stronger association of the SLC6A4-DRD2 interaction with DT (P < 0.0001), which remained significant after Bonferroni correction. Results reveal decreased likelihood of DT in alcoholics that carry the DRD2 rs6276 G allele and SLC6A4 LL genotype. This study provides the first evidence to implicate the interaction between serotonin and dopamine neurotransmission in the etiology of DT. Replication is necessary to verify this potentially important finding. PMID- 20002021 TI - Atypical development of behavioural sensitization to 3,4 methylenedioxymethamphetamine (MDMA, 'Ecstasy') in adolescent rats and its expression in adulthood: role of the MDMA chirality. AB - Despite the great popularity of 3,4-methylenedioxymethamphetamine (MDMA, Ecstasy) as a drug of abuse, not much is known about the detailed mechanisms of the acute and subchronic effects of the drug. There is especially a lack of information about the distinct behavioural effects of its optical isomers (enantiomers) R- and S-MDMA compared with the racemic RS-MDMA. For this purpose, adolescent rats were repetitively treated during two treatment stages (stage 1: days 1-10; stage 2: days 15, 17, 19) with RS-MDMA (5 or 10 mg/kg) or each of the respective enantiomers (5 mg/kg). The repeated treatment started on postnatal day (PND) 32 and locomotor activity was measured on each day by means of a photobeam-equipped activity box system. RS-MDMA or S-MDMA administration led acutely to massive hyperlocomotion and subchronically, to the development of behavioural sensitization after a short habituation period. R-MDMA was free of hyperactivating effects and even decreased locomotor behaviour upon repeated treatment. Nevertheless, co-administration of R-MDMA increased the hyperactivity of S-MDMA and made the S-MDMA induced behavioural sensitization state-dependent. The animals pre-treated with R-MDMA showed a sensitized response in adulthood when tested with RS-MDMA. Our results indicated that even in the absence of substantial neurotoxicity, both MDMA enantiomers can lead to long-term changes in brain circuitry and concomitant behavioural changes when repeatedly administered in adolescence. The sensitization development was most pronounced in the animals treated with S- and RS-MDMA; the animals with R-MDMA did not develop sensitization under repeated treatment but expressed a sensitized response when challenged with RS-MDMA. PMID- 20002023 TI - Mechanisms of respiratory insufficiency induced by methadone overdose in rats. AB - Methadone may cause respiratory depression. We aimed to understand methadone related effects on ventilation as well as each opioid-receptor (OR) role. We studied the respiratory effects of intraperitoneal methadone at 1.5, 5, and 15 mg/kg (corresponding to 80% of the lethal dose-50%) in rats using arterial blood gases and plethysmography. OR antagonists, including intravenous 10 mg/kg naloxonazine at 5 minutes (mu-OR antagonist), subcutaneous 30 mg/kg-naloxonazine at 24 hours (micro1-OR antagonist), 3 mg/kg-naltrindole at 45 minutes (delta-OR antagonist) and 5 mg/kg-Nor-binaltorphimine at 6 hours (kappa-OR antagonist) were pre-administered. Plasma concentrations of methadone enantiomers were measured using high-performance liquid chromatography coupled to mass-spectrometry. Methadone dose-dependent inspiratory time (T(I)) increase tended to be linear. Respiratory depression was observed only at 15 mg/kg and characterized by an increase in expiratory time (T(E)) resulting in hypoxemia and respiratory acidosis. Intravenous naloxonazine completely reversed all methadone-related effects on ventilation, while subcutaneous naloxonazine reduced its effects on pH (P < 0.05), PaCO(2) (P < 0.01) and T(E) (P < 0.001) but only partially on T(I) (P < 0.001). Naltrindole reduced methadone-related effects on T(E) (P < 0.001). Nor binaltorphimine increased methadone-related effects on pH and PaO(2) (P < 0.05) Respiratory effects as a function of plasma R-methadone concentrations showed a decrease in PaO(2) (EC(50): 1.14 microg/ml) at lower concentrations than those necessary for PaCO(2) increase (EC(50): 3.35 microg/ml). Similarly, increased T(I) (EC(50): 0.501 microg/ml) was obtained at lower concentrations than those for T(E) (EC(50): 4.83 microg/ml). Methadone-induced hypoxemia is caused by mu ORs and modulated by kappa-ORs. Additionally, methadone-induced increase in T(E) is caused by mu1- and delta-opioid receptors while increase in T(I) is caused by mu-ORs. PMID- 20002024 TI - Phosphatidylethanol: normalization during detoxification, gender aspects and correlation with other biomarkers and self-reports. AB - Phosphatidylethanol (PEth) is a direct ethanol metabolite, and has recently attracted attention as biomarker of ethanol intake. The aims of the current study are: (1) to characterize the normalization time of PEth in larger samples than previously conducted; (2) to elucidate potential gender differences; and (3) to report the correlation of PEth with other biomarkers and self-reported alcohol consumption. Fifty-seven alcohol-dependent patients (ICD 10 F 10.25; 9 females, 48 males) entering medical detoxification at three study sites were enrolled. The study sample was comprised of 48 males and 9 females, with mean age 43.5. Mean gamma glutamyl transpeptidase (GGT) was 209.61 U/l, average mean corpuscular volume (MCV) was 97.35 fl, mean carbohydrate deficient transferrin (%CDT) was 8.68, and mean total ethanol intake in the last 7 days was 1653 g. PEth was measured in heparinized whole blood with a high-pressure liquid chromatography method, while GGT, MCV and %CDT were measured using routine methods. PEth levels at day 1 of detoxification ranged between 0.63 and 26.95 micromol/l (6.22 mean, 4.70 median, SD 4.97). There were no false negatives at day 1. Sensitivities for the other biomarkers were 40.4% for MCV, 73.1% for GGT and 69.2% for %CDT, respectively. No gender differences were found for PEth levels at any time point. Our data suggest that PEth is (1) a suitable intermediate term marker of ethanol intake in both sexes; and (2) sensitivity is extraordinary high in alcohol dependent patients. The results add further evidence to the data that suggest that PEth has potential as a candidate for a sensitive and specific biomarker, which reflects longer-lasting intake of higher amounts of alcohol and seemingly has the above mentioned certain advantages over traditional biomarkers. PMID- 20002022 TI - Mice lacking Gad2 show altered behavioral effects of ethanol, flurazepam and gabaxadol. AB - Gamma-aminobutyric acid (GABA) is synthesized in brain by two isoforms of glutamic acid decarboxylase (Gad), Gad1 and Gad2. Gad1 provides most of the GABA in brain, but Gad2 can be rapidly activated in times of high GABA demand. Mice lacking Gad2 are viable whereas deletion of Gad1 is lethal. We produced null mutant mice for Gad2 on three different genetic backgrounds: predominantly C57BL/6J and one or two generations of backcrossing to 129S1/SvimJ (129N1, 129N2). We used these mice to determine if actions of alcohol are regulated by synthesis of GABA from this isoform. We also studied behavioral responses to a benzodiazepine (flurazepam) and a GABAA receptor agonist (gabaxadol). Deletion of Gad2 increased ethanol palatability and intake and slightly reduced the severity of ethanol-induced withdrawal, but these effects depended strongly on genetic background. Mutant mice on the 129N2 background showed the above three ethanol behavioral phenotypes, but the C57BL/6J inbred background did not show any of these phenotypes. Effects on ethanol consumption also depended on the test as the mutation did not alter consumption in limited access models. Deletion of Gad2 reduced the effect of flurazepam on motor incoordination and increased the effect of extrasynaptic GABAA receptor agonist gabaxadol without changing the duration of loss of righting reflex produced by these drugs. These results are consistent with earlier proposals that deletion of Gad2 (on 129N2 background) reduces synaptic GABA but also suggest changes in extrasynaptic receptor function. PMID- 20002025 TI - The safety of intravenous fluorescein for confocal laser endomicroscopy in the gastrointestinal tract. AB - BACKGROUND: Confocal laser endomicroscopy (CLE) is rapidly emerging as a valuable tool for gastrointestinal endoscopic imaging. Fluorescent contrast agents are used to optimize imaging with CLE, and intravenous fluorescein is the most widely used contrast agent. Fluorescein is FDA-cleared for diagnostic angiography of the retina. For these indications, the safety profile of fluorescein has been well documented; however, to date, fluorescein is not cleared for use with CLE. AIMS: To estimate the rate of serious and total adverse events attributable to intravenous fluorescein when used for gastrointestinal CLE. METHODS: We performed a cross sectional survey of 16 International Academic Medical Centres with active research protocols in CLE that involved intravenous fluorescein. Centres using i.v. fluorescein for CLE who were actively monitored for adverse events were included. RESULTS: Sixteen centres performed 2272 gastrointestinal CLE procedures. The most common dose of contrast agent was 2.5-5 mL of 10% sodium fluorescein. No serious adverse events were reported. Mild adverse events occurred in 1.4% of individuals, including nausea/vomiting, transient hypotension without shock, injection site erythema, diffuse rash and mild epigastric pain. The limitation is that only immediate post procedure events were actively monitored. CONCLUSIONS: Use of intravenous fluorescein for gastrointestinal CLE appears to be safe with few acute complications. PMID- 20002026 TI - Infliximab safety profile and long-term applicability in inflammatory bowel disease: 9-year experience in clinical practice. AB - BACKGROUND: Most available data on infliximab therapy come from large, short term, pivotal RCTs and concerns about long-term safety profile still remain. AIM: To evaluate the long-term safety profile of infliximab in inflammatory bowel disease (IBD) in a clinical practice setting. METHODS: Since 1999, all IBD patients treated with infliximab were registered and clinical outcomes prospectively recorded up to March 2008, loss of follow-up or patient's death. Infliximab regimens and preventive measures were in accordance with the prevalent guidelines or with the manufacturer's recommendations. RESULTS: One hundred fifty two patients were included (121 Crohn's disease, 24 ulcerative colitis, 7 indeterminate colitis), with a median of 5 infliximab infusions (IQR 3-8) and 87% of patients received at least three infusions. Seventy-nine per cent of them received concomitant immunomodulators and 70% were pre-medicated with hydrocortisone from the first infusion. After a median follow-up of 142 weeks, 13% presented infusion reactions, 13% viral or bacterial infections and two patients developed neoplasia. The mortality rate was 2.6% (four patients). CONCLUSIONS: Infliximab therapy is safe when the recommended preventive measures are implemented, with a rate of serious adverse events less than 10%. No new safety signals were found. PMID- 20002030 TI - In vitro comparative studies on pancreatic enzyme preparations. PMID- 20002027 TI - Review article: the modern management of hepatic encephalopathy. AB - BACKGROUND; Hepatic encephalopathy, both overt and minimal, forms a continuum of cognitive change in cirrhosis. Strategies to diagnose and treat hepatic encephalopathy have evolved considerably. AIM: To examine the updated diagnostic and treatment strategies for hepatic encephalopathy. METHODS: Techniques for the clinical, psychometric and neurophysiological evaluation of hepatic encephalopathy are reviewed. The methods reviewed include pure clinical scales (West-Haven), psychometric tests (PSE-syndrome test), neurophysiological tests (EEG, Critical flicker frequency, CFF) and computerized tests (Inhibitory control test, ICT). RESULTS: Clinical scales are limited, whereas psychometric tests (specifically PSE-syndrome test), CFF and ICT can be used to diagnose minimal hepatic encephalopathy. However, there is no single test that can capture the entire spectrum of cognitive impairment. Treatment options and goals depend on the acuity of hepatic encephalopathy. In-patient management should concentrate on supportive care, precipitating factor reversal and lactulose and/or rifaximin therapy. Out-patient therapy should aim to prevent recurrences, and both lactulose and rifaximin have evidence to support their use. CONCLUSIONS: Diagnostic techniques for hepatic encephalopathy range from simple scales to sophisticated tools. Treatment options depend on the stage of hepatic encephalopathy. The future challenge is to evaluate cognitive function as a continuum with clinically relevant outcomes and to develop well-tolerated and inexpensive treatments for hepatic encephalopathy. PMID- 20002031 TI - Applicability of the reported prevalence of bile salt malabsorption in irritable bowel. PMID- 20002033 TI - Paroxysmal nocturnal haemoglobinuria clones are part of the immune dysregulation in hepatitis-associated aplastic anaemia. PMID- 20002036 TI - Chernobyl catastrophe. Foreword. PMID- 20002037 TI - Consequences of the Chernobyl catastrophe. Preface. PMID- 20002038 TI - Introduction: the difficult truth about Chernobyl. PMID- 20002040 TI - 1. Chernobyl contamination through time and space. AB - Radioactive contamination from the Chernobyl meltdown spread over 40% of Europe (including Austria, Finland, Sweden, Norway, Switzerland, Romania, Great Britain, Germany, Italy, France, Greece, Iceland, Slovenia) and wide territories in Asia (including Turkey, Georgia, Armenia, Emirates, China), northern Africa, and North America. Nearly 400 million people resided in territories that were contaminated with radioactivity at a level higher than 4 kBq/m(2) (0.11 Ci/km(2)) from April to July 1986. Nearly 5 million people (including, more than 1 million children) still live with dangerous levels of radioactive contamination in Belarus, Ukraine, and European Russia. Claims that the Chernobyl radioactive fallout adds "only 2%" to the global radioactive background overshadows the fact that many affected territories had previously dangerously high levels of radiation. Even if the current level is low, there was high irradiation in the first days and weeks after the Chernobyl catastrophe. There is no reasonable explanation for the fact that the International Atomic Energy Agency and the World Health Organization (Chernobyl Forum, 2005) have completely neglected the consequences of radioactive contamination in other countries, which received more than 50% of the Chernobyl radionuclides, and addressed concerns only in Belarus, Ukraine, and European Russia. PMID- 20002042 TI - 2. Chernobyl's public health consequences. AB - Problems complicating a full assessment of the effects from Chernobyl included official secrecy and falsification of medical records by the USSR for the first 3.5 years after the catastrophe and the lack of reliable medical statistics in Ukraine, Belarus, and Russia. Official data concerning the thousands of cleanup workers (Chernobyl liquidators) who worked to control the emissions are especially difficult to reconstruct. Using criteria demanded by the International Atomic Energy Agency (IAEA), the World Health Organization (WHO), and the United Nations Scientific Committee on the Effects of Atomic Radiation (UNSCEAR) resulted in marked underestimates of the number of fatalities and the extent and degree of sickness among those exposed to radioactive fallout from Chernobyl. Data on exposures were absent or grossly inadequate, while mounting indications of adverse effects became more and more apparent. Using objective information collected by scientists in the affected areas--comparisons of morbidity and mortality in territories characterized by identical physiography, demography, and economy, which differed only in the levels and spectra of radioactive contamination--revealed significant abnormalities associated with irradiation, unrelated to age or sex (e.g., stable chromosomal aberrations), as well as other genetic and nongenetic pathologies. PMID- 20002043 TI - 3. General morbidity, impairment, and disability after the chernobyl catastrophe. AB - In all cases when comparing the territories heavily contaminated by Chernobyl's radionuclides with less contaminated areas that are characterized by a similar economy, demography, and environment, there is a marked increase in general morbidity in the former. Increased numbers of sick and weak newborns were found in the heavily contaminated territories in Belarus, Ukraine, and European Russia. PMID- 20002044 TI - 4. Accelerated aging as a consequence of the Chernobyl catastrophe. AB - Accelerated aging is one of the well-known consequences of exposure to ionizing radiation. This phenomenon is apparent to a greater or lesser degree in all of the populations contaminated by the Chernobyl radionuclides. PMID- 20002045 TI - 5. Nonmalignant diseases after the Chernobyl catastrophe. AB - This section describes the spectrum and the scale of the nonmalignant diseases that have been found among exposed populations. Adverse effects as a result of Chernobyl irradiation have been found in every group that has been studied. Brain damage has been found in individuals directly exposed--liquidators and those living in the contaminated territories, as well as in their offspring. Premature cataracts; tooth and mouth abnormalities; and blood, lymphatic, heart, lung, gastrointestinal, urologic, bone, and skin diseases afflict and impair people, young and old alike. Endocrine dysfunction, particularly thyroid disease, is far more common than might be expected, with some 1,000 cases of thyroid dysfunction for every case of thyroid cancer, a marked increase after the catastrophe. There are genetic damage and birth defects especially in children of liquidators and in children born in areas with high levels of radioisotope contamination. Immunological abnormalities and increases in viral, bacterial, and parasitic diseases are rife among individuals in the heavily contaminated areas. For more than 20 years, overall morbidity has remained high in those exposed to the irradiation released by Chernobyl. One cannot give credence to the explanation that these numbers are due solely to socioeconomic factors. The negative health consequences of the catastrophe are amply documented in this chapter and concern millions of people. PMID- 20002046 TI - 6. Oncological diseases after the Chernobyl catastrophe. AB - The most recent forecast by international agencies predicted there would be between 9,000 and 28,000 fatal cancers between 1986 and 2056, obviously underestimating the risk factors and the collective doses. On the basis of I-131 and Cs-137 radioisotope doses to which populations were exposed and a comparison of cancer mortality in the heavily and the less contaminated territories and pre- and post-Chernobyl cancer levels, a more realistic figure is 212,000 to 245,000 deaths in Europe and 19,000 in the rest of the world. High levels of Te-132, Ru 103, Ru-106, and Cs-134 persisted months after the Chernobyl catastrophe and the continuing radiation from Cs-137, Sr-90, Pu, and Am will generate new neoplasms for hundreds of years. PMID- 20002047 TI - 7. Mortality after the Chernobyl catastrophe. AB - A detailed study reveals that 3.8-4.0% of all deaths in the contaminated territories of Ukraine and Russia from 1990 to 2004 were caused by the Chernobyl catastrophe. The lack of evidence of increased mortality in other affected countries is not proof of the absence of effects from the radioactive fallout. Since 1990, mortality among liquidators has exceeded the mortality rate in corresponding population groups. From 112,000 to 125,000 liquidators died before 2005--that is, some 15% of the 830,000 members of the Chernobyl cleanup teams. The calculations suggest that the Chernobyl catastrophe has already killed several hundred thousand human beings in a population of several hundred million that was unfortunate enough to live in territories affected by the fallout. The number of Chernobyl victims will continue to grow over many future generations. PMID- 20002048 TI - Conclusion to Chapter II. PMID- 20002049 TI - Chapter III. Consequences of the Chernobyl catastrophe for the environment. PMID- 20002050 TI - 8. Atmospheric, water, and soil contamination after Chernobyl. AB - Air particulate activity over all of the Northern Hemisphere reached its highest levels since the termination of nuclear weapons testing--sometimes up to 1 million times higher than before the Chernobyl contamination. There were essential changes in the ionic, aerosol, and gas structure of the surface air in the heavily contaminated territories, as measured by electroconductivity and air radiolysis. Many years after the catastrophe aerosols from forest fires have dispersed hundreds of kilometers away. The Chernobyl radionuclides concentrate in sediments, water, plants, and animals, sometimes 100,000 times more than the local background level. The consequences of such a shock on aquatic ecosystems is largely unclear. Secondary contamination of freshwater ecosystems occurs as a result of Cs-137 and Sr-90 washout by the high waters of spring. The speed of vertical migration of different radionuclides in floodplains, lowland moors, peat bogs, etc., is about 2-4 cm/year. As a result of this vertical migration of radionuclides in soil, plants with deep root systems absorb them and carry the ones that are buried to the surface again. This transfer is one of the important mechanisms, observed in recent years, that leads to increased doses of internal irradiation among people in the contaminated territories. PMID- 20002051 TI - 9. Chernobyl's radioactive impact on flora. AB - Plants and mushrooms accumulate the Chernobyl radionuclides at a level that depends upon the soil, the climate, the particular biosphere, the season, spotty radioactive contamination, and the particular species and populations (subspecies, cultivars), etc. Each radionuclide has its own accumulation characteristics (e. g., levels of accumulation for Sr-90 are much higher than for Cs-137, and a thousand times less than that for Ce-144). Coefficients of accumulation and transition ratios vary so much in time and space that it is difficult, if not impossible, to predict the actual levels of Cs-137, Sr-90, Pu 238, Pu-239, Pu-240, and Am-241 at each place and time and for each individual plant or fungus. Chernobyl irradiation has caused structural anomalies and tumorlike changes in many plant species. Unique pathologic complexes are seen in the Chernobyl zone, such as a high percentage of anomalous pollen grains and spores. Chernobyl's irradiation has led to genetic disorders, sometimes continuing for many years, and it appears that it has awakened genes that have been silent over a long evolutionary time. PMID- 20002052 TI - 10. Chernobyl's radioactive impact on fauna. AB - The radioactive shock when the Chernobyl reactor exploded in 1986 combined with chronic low-dose contamination has resulted in morphologic, physiologic, and genetic disorders in every animal species that has been studied--mammals, birds, amphibians, fish, and invertebrates. These populations exhibit a wide variety of morphological deformities not found in other populations. Despite reports of a "healthy" environment in proximity to Chernobyl for rare species of birds and mammals, the presence of such wildlife is likely the result of immigration and not from locally sustained populations. Twenty-three years after the catastrophe levels of incorporated radionuclides remain dangerously high for mammals, birds, amphibians, and fish in some areas of Europe. Mutation rates in animal populations in contaminated territories are significantly higher and there is transgenerational genomic instability in animal populations, manifested in adverse cellular and systemic effects. Long-term observations of both wild and experimental animal populations in the heavily contaminated areas show significant increases in morbidity and mortality that bear a striking resemblance to changes in the health of humans--increased occurrence of tumor and immunodeficiencies, decreased life expectancy, early aging, changes in blood and the circulatory system, malformations, and other factors that compromise health. PMID- 20002053 TI - 11. Chernobyl's radioactive impact on microbial biota. AB - Of the few microorganisms that have been studied, all underwent rapid changes in the areas heavily contaminated by Chernobyl. Organisms such as tuberculosis bacilli; hepatitis, herpes, and tobacco mosaic viruses; cytomegalovirus; and soil micromycetes and bacteria were activated in various ways. The ultimate long-term consequences for the Chernobyl microbiologic biota may be worse than what we know today. Compared to humans and other mammals, the profound changes that take place among these small live organisms with rapid reproductive turnover do not bode well for the health and survival of other species. PMID- 20002054 TI - Conclusion to Chapter III. PMID- 20002055 TI - Chapter IV. Radiation protection after the Chernobyl catastrophe. PMID- 20002056 TI - 12. Chernobyl's radioactive contamination of food and people. AB - In many European countries levels of I-131, Cs-134/137, Sr-90, and other radionuclides in milk, dairy products, vegetables, grains, meat, and fish increased drastically (sometimes as much as 1,000-fold) immediately after the catastrophe. Up until 1991 the United States imported food products with measurable amounts of Chernobyl radioactive contamination, mostly from Turkey, Italy, Austria, West Germany, Greece, Yugoslavia, Hungary, Sweden, and Denmark. These products included juices, cheeses, pasta, mushrooms, hazelnuts, sage, figs, tea, thyme, juniper, caraway seeds, and apricots. In Gomel, Mogilev, and Brest provinces in Belarus 7-8% of milk and 13-16% of other food products from small farms exceeded permissible levels of Cs-137, even as recently as 2005-2007. As of 2000, up to 90% of the wild berries and mushrooms exceeded permissible levels of Cs-137 in Rovno and Zhytomir provinces, Ukraine. Owing to weight and metabolic differences, a child's radiation exposure is 3-5 times higher than that of an adult on the same diet. From 1995 to 2007, up to 90% of the children from heavily contaminated territories of Belarus had levels of Cs-137 accumulation higher than 15-20 Bq/kg, with maximum levels of up to 7,300 Bq/kg in Narovlya District, Gomel Province. Average levels of incorporated Cs-137 and Sr-90 in the heavily contaminated territories of Belarus, Ukraine, and European Russia did not decline, but rather increased from 1991 to 2005. Given that more than 90% of the current radiation fallout is due to Cs-137, with a half-life of about 30 years, we know that the contaminated areas will be dangerously radioactive for roughly the next three centuries. PMID- 20002057 TI - 13. Decorporation of Chernobyl radionuclides. AB - Tens of thousands of Chernobyl children (mostly from Belarus) annually leave to receive treatment and health care in other countries. Doctors from many countries gratuitously work in the Chernobyl contaminated territories, helping to minimize the consequences of this most terrible technologic catastrophe in history. But the scale and spectrum of the consequences are so high, that no country in the world can cope alone with the long-term consequences of such a catastrophe as Chernobyl. The countries that have suffered the most, especially Ukraine and Belarus, extend gratitude for the help that has come through the United Nations and other international organizations, as well as from private funds and initiatives. Twenty-two years after the Chernobyl releases, the annual individual dose limit in heavily contaminated territories of Belarus, Ukraine, and European Russia exceed 1 mSv/year just because of the unavoidable consumption of locally contaminated products. The 11-year experience of the BELRAD Institute shows that for effective radiation protection it is necessary to establish the interference level for children at 30% of the official dangerous limit (i.e., 15-20 Bq/kg). The direct whole body counting measurements of Cs-137 accumulation in the bodies of inhabitants of the heavily contaminated Belarussian region shows that the official Dose Catalogue underestimates the annual dose burdens by three to eight times. For practical reasons the curative-like use of apple-pectin food additives might be especially helpful for effective decorporation of Cs-137. From 1996 to 2007 a total of more than 160,000 Belarussian children received pectin food additives during 18 to 25 days of treatment (5 g twice a day). As a result, levels of Cs-137 in children's organs decreased after each course of pectin additives by an average of 30 to 40%. Manufacture and application of various pectin-based food additives and drinks (using apples, currants, grapes, sea seaweed, etc.) is one of the most effective ways for individual radioprotection (through decorporation) under circumstances where consumption of radioactively contaminated food is unavoidable. PMID- 20002058 TI - 14. Protective measures for activities in Chernobyl's radioactively contaminated territories. AB - Owing to internally absorbed radionuclides, radiation levels for individuals living in the contaminated territories of Belarus, Ukraine, and Russia have been increasing steadily since 1994. Special protective measures in connection with agriculture, forestry, hunting, and fishing are necessary to protect the health of people in all the radioactively contaminated territories. Among the measures that have proven to be effective in reducing levels of incorporated radionuclides in meat production are food additives with ferrocyanides, zeolites, and mineral salts. Significant decreases in radionuclide levels in crops are achieved using lime/Ca as an antagonist of Sr-90, K fertilizers as antagonists of Cs-137, and phosphoric fertilizers that form a hard, soluble phosphate with Sr-90. Disk tillage and replowing of hayfields incorporating applications of organic and mineral fertilizers reduces the levels of Cs-137 and Sr-90 three- to fivefold in herbage grown in mineral soils. Among food technologies to reduce radionuclide content are cleaning cereal seeds, processing potatoes into starch, processing carbohydrate-containing products into sugars, and processing milk into cream and butter. There are several simple cooking techniques that decrease radionuclides in foodstuffs. Belarus has effectively used some forestry operations to create "a live partition wall," to regulate the redistribution of radionuclides into ecosystems. All such protective measures will be necessary in many European territories for many generations. PMID- 20002059 TI - 15. Consequences of the Chernobyl catastrophe for public health and the environment 23 years later. AB - More than 50% of Chernobyl's radionuclides were dispersed outside of Belarus, Ukraine, and European Russia and caused fallout as far away as North America. In 1986 nearly 400 million people lived in areas radioactively contaminated at a level higher than 4 kBq/m(2) and nearly 5 million individuals are still being exposed to dangerous contamination. The increase in morbidity, premature aging, and mutations is seen in all the contaminated territories that have been studied. The increase in the rates of total mortality for the first 17 years in European Russia was up to 3.75% and in Ukraine it was up to 4.0%. Levels of internal irradiation are increasing owing to plants absorbing and recycling Cs-137, Sr-90, Pu, and Am. During recent years, where internal levels of Cs-137 have exceeded 1 mSv/year, which is considered "safe," it must be lowered to 50 Bq/kg in children and to 75 Bq/kg in adults. Useful practices to accomplish this include applying mineral fertilizers on agricultural lands, K and organosoluble lignin on forestlands, and regular individual consumption of natural pectin enterosorbents. Extensive international help is needed to provide radiation protection for children, especially in Belarus, where over the next 25 to 30 years radionuclides will continue to contaminate plants through the root layers in the soil. Irradiated populations of plants and animals exhibit a variety of morphological deformities and have significantly higher levels of mutations that were rare prior to 1986. The Chernobyl zone is a "black hole": some species may persist there only via immigration from uncontaminated areas. PMID- 20002060 TI - Conclusion to Chapter IV. PMID- 20002061 TI - Dose-dependent adsorptive capacity of activated charcoal for gastrointestinal decontamination of a simulated paracetamol overdose in human volunteers. AB - The amount of activated charcoal needed to treat drug overdoses has arbitrarily been set at a charcoal-drug ratio of 10:1. Recent in vitro studies have shown a larger adsorptive capacity for activated charcoal when used in a model of paracetamol overdose. In the present study, we investigated whether this reserve capacity exists in vivo. This is clinically relevant in cases of large overdoses or if the full standard dose of 50 g activated charcoal cannot be administered. We performed a randomized, cross-over study (n = 16). One hour after a standard breakfast, 50 mg/kg paracetamol was administered, followed 1 hr later by an activated charcoal-Water slurry containing 50 (control), 25 or 5 g activated charcoal. The areas under the serum concentration-time curve (AUC) for paracetamol were used to estimate the efficacy of each activated charcoal dose. The AUC of the 25-g dose was found to be of similar size compared to the control, although statistics were weak. The AUC of the 5-g dose was 59% larger than the AUC of the 50-g dose (p = 0.0003). The terminal elimination half-life (t(1/2)) of paracetamol was 1.6 (CI 1.4-2.0) and 1.9 (CI 1.5-2.4) hr for 50 and 25 g, respectively (NS), and 2.5 (CI 1.8-3.0) hr for the 5-g dose (p = 0.003). The decrease in t(1/2) of paracetamol for the two larger activated charcoal doses indicates a possible effect of activated charcoal on paracetamol clearance and warrants further investigation. The large adsorptive reserve capacity of activated charcoal in vitro could not be reproduced for the smallest dose of activated charcoal. An activated charcoal-drug ratio of 10:1 is therefore still recommendable. PMID- 20002062 TI - A synthetic analogue of 20-HETE, 5,14-HEDGE, reverses endotoxin-induced hypotension via increased 20-HETE levels associated with decreased iNOS protein expression and vasodilator prostanoid production in rats. AB - Nitric oxide (NO) produced by inducible NO synthase (iNOS) is responsible for endotoxin (ET)-induced hypotension and vascular hyporeactivity and plays a major contributory role in the multiorgan failure. Endotoxic shock is also associated with an increase in vasodilator prostanoids as well as a decrease in endothelial NO synthase (eNOS) and cytochrome P450 4A protein expression, and production of a vasoconstrictor arachidonic acid product, 20-hydroxyeicosatetraenoic acid (20 HETE). The aim of this study was to investigate the effects of a synthetic analogue of 20-HETE, N-[20-hydroxyeicosa-5(Z),14(Z)-dienoyl]glycine (5,14-HEDGE), on the ET-induced changes in eNOS, iNOS and heat shock protein 90 (hsp90) expression as well as 20-HETE and vasodilator prostanoid (6-keto-PGF(1alpha) and PGE(2)) production. ET-induced fall in blood pressure and rise in heart rate were associated with an increase in iNOS protein expression and a decrease in eNOS protein expression in heart, thoracic aorta, kidney and superior mesenteric artery. ET did not change hsp90 protein expression in the tissues. ET-induced changes in eNOS and iNOS protein expression were associated with increased 6-keto PGF(1alpha) and PGE(2) levels and a decrease in 20-HETE levels, in the serum and kidney. These effects of ET on the iNOS protein expression and 6-keto PGF(1alpha), PGE(2) and 20-HETE levels were prevented by 5,14-HEDGE. Furthermore, a competitive antagonist of vasoconstrictor effects of 20-HETE, 20-hydroxyeicosa 6(Z),15(Z)-dienoic acid, prevented the effects of 5,14-HEDGE on the ET-induced changes in systemic and renal levels of these prostanoids and 20-HETE. These data are consistent with the view that an increase in systemic and renal 20-HETE levels associated with a decrease in iNOS protein expression and vasodilator prostanoid production contributes to the effect of 5,14-HEDGE to prevent the hypotension during rat endotoxemia. PMID- 20002063 TI - Effect of carbamazepine and lamotrigine on cognitive function and oxidative stress in brain during chemical epileptogenesis in rats. AB - The present study assessed the effect of carbamazepine and lamotrigine on cognitive function and oxidative stress in brain during chemically induced epileptogenesis in rats. Epileptogenesis was induced by administration of pentylenetetrazole (30 mg/kg, s.c.) on alternate days (three times/week) for 9-11 weeks or until stage 4 of seizure score was achieved. The neurobehavioural parameters used for cognitive assessment were step-down latency in continuous avoidance apparatus and transfer latency in elevated plus maze test paradigm. Carbamazepine and lamotrigine were administered intraperitoneally in doses of 60 mg/kg and 25 mg/kg, respectively, according to the groups, once a day for 11 weeks. Oxidative stress was assessed in isolated homogenized whole brain samples and estimated for the levels of malondialdehyde, reduced glutathione, catalase and superoxide dismutase. The results showed that lamotrigine did not produce any change in cognitive function, while carbamazepine produced cognitive dysfunction. Cognitive decline seen in the carbamazepine-treated pentylenetetrazole-kindled group was also associated with increased oxidative stress. Lamotrigine treatment had no effect on oxidative stress parameters alone, while it significantly decreased oxidative stress in the pentylenetetrazole-kindled group as compared to the pentylenetetrazole-kindled carbamazepine-treated group. PMID- 20002064 TI - Predicting drug disposition via application of a Biopharmaceutics Drug Disposition Classification System. AB - A Biopharmaceutics Drug Disposition Classification System (BDDCS) was proposed to serve as a basis for predicting the importance of transporters in determining drug bioavailability and disposition. BDDCS may be useful in predicting: routes of drug elimination; efflux and absorptive transporters effects on oral absorption; when transporter-enzyme interplay will yield clinically significant effects (e.g. low drug bioavailability and drug-drug interactions); and transporter effects on post-absorptive systemic drug levels following oral and i.v. dosing. For highly soluble, highly permeable Class 1 compounds, metabolism is the major route of elimination and transporter effects on drug bioavailability and hepatic disposition are negligible. In contrast for the poorly permeable Class 3 and 4 compounds, metabolism only plays a minor role in drug elimination. Uptake transporters are major determinants of drug bioavailability for these poorly permeable drugs and both uptake and efflux transporters could be important for drug elimination. Highly permeable, poorly soluble, extensively metabolized Class 2 compounds present the most complicated relationship in defining the impact of transporters due to a marked transporter-enzyme interplay. Uptake transporters are unimportant for Class 2 drug bioavailability, (ensure space after,) but can play a major role in hepatic and renal elimination. Efflux transporters have major effects on drug bioavailability, absorption, metabolism and elimination of Class 2 drugs. It is difficult to accurately characterize drugs in terms of the high permeability criteria, i.e. > or =90% absorbed. We suggest that extensive metabolism may substitute for the high permeability characteristic, and that BDDCS using elimination criteria may provide predictability in characterizing drug disposition profiles for all classes of compounds. PMID- 20002065 TI - Regulation of low-density lipoprotein receptor and 3-hydroxy-3-methylglutaryl coenzyme A reductase expression by Zingiber officinale in the liver of high-fat diet-fed rats. AB - Zingiber officinale has been used to control lipid disorders and reported to possess remarkable cholesterol-lowering activity in experimental hyperlipidaemia. In the present study, the effect of a characterized and standardized extract of Zingiber officinale on the hepatic lipid levels as well as on the hepatic mRNA and protein expression of low-density lipoprotein (LDL) receptor and 3-hydroxy-3 methylglutaryl coenzyme A (HMG-CoA) reductase was investigated in a high-fat diet fed rat model. Rats were treated with an ethanol extract of Zingiber officinale (400 mg/kg) extract along with a high-fat diet for 6 weeks. The extract of Zingiber officinale significantly decreased hepatic triglyceride and tended to decrease hepatic cholesterol levels when administered over 6 weeks to the rats fed a high-fat diet. We found that in parallel, the extract up-regulated both LDL receptor mRNA and protein level and down-regulated HMG-CoA reductase protein expression in the liver of these rats. The metabolic control of body lipid homeostasis is in part due to enhanced cholesterol biosynthesis and reduced expression of LDL receptor sites following long-term consumption of high-fat diets. The present results show restoration of transcriptional and post transcriptional changes in low-density lipoprotein and HMG CoA reductase by Zingiber officinale administration with a high-fat diet and provide a rational explanation for the effect of ginger in the treatment of hyperlipidaemia. PMID- 20002066 TI - Exercise and supraphysiological dose of nandrolone decanoate increase apoptosis in spermatogenic cells. AB - Anabolic-androgenic steroids are used at high doses by athletes for improving athletic ability, physical appearance and muscle mass. Unfortunately, the abuse of these agents has significantly increased. It has been established that exercise and high doses of anabolic-androgenic steroids may influence the hypothalamic-pituitary-gonadal axis, which can in turn affect testicular apoptosis. However, the effect of the combination of exercise and high dose of anabolic-androgenic steroids on testicular apoptosis is not known. We investigated the combined effects of exercise and high doses of nandrolone decanoate on apoptosis in the spermatogenic cell lineage. Five groups of male Wistar strain albino rats were treated as follows for 8 weeks: solvent of nandrolone decanoate (peanut oil) as a vehicle (Sham); nandrolone decanoate (10 mg/kg/weekly) (nandrolone decanoate); exercise (1 hr/day, 5 days a week) (exercise); nandrolone decanoate (10 mg/kg/weekly) and exercise (1 hr/day, 5 days a week) (nandrolone decanoate exercise); and sedentary control without any injection or exercise (Control). Apoptosis in the male germ line was characterized by TUNEL, caspase-3 assay and transmission electron microscopy. The weights of the testis and accessory sex organs, as well as sperm parameters significantly decreased in the experimental groups relative to the sham and control groups (p < or = 0.05). Germ cell apoptosis and a significant decrease in the number of germ cell layers in nandrolone decanoate exercise-treated testes were observed (p < or = 0.05). Exercise training seems to increase the extent of apoptotic changes caused by supraphysiological dose of nandrolone decanoate in rats, which in turn affects fertility. PMID- 20002067 TI - Hypotensive and vasorelaxant effects of citronellol, a monoterpene alcohol, in rats. AB - Citronellol is an essential oil constituent from the medicinal plants Cymbopogon citratus, Cymbopogon winterianus and Lippia alba which are thought to possess antihypertensive properties. Citronellol-induced cardiovascular effects were evaluated in this study. In rats, citronellol (1-20 mg/kg, i.v.) induced hypotension, which was not affected by pre-treatment with atropine, hexamethonium, N(omega)-nitro-L-arginine methyl ester hydrochloride or indomethacin, and tachycardia, which was only attenuated by pre-treatment with atropine and hexamethonium. These responses were less than those obtained for nifedipine, a reference drug. In intact rings of rat mesenteric artery pre contracted with 10 microM phenylephrine, citronellol induced relaxations (pD(2) = 0.71 +/- 0.11; E(max) = 102 +/- 5%; n = 6) that were not affected by endothelium removal, after tetraethylamonium in rings without endothelium pre-contracted with KCl 80 mM. Citronellol strongly antagonized (maximal inhibition = 97 +/- 4%; n = 6) the contractions induced by CaCl(2) (10(-6) to 3 x 10(-3 )M) and did not induce additional effects on the maximal response of nifedipine (10 microM). Finally, citronellol inhibited the contractions induced by 10 microM phenylephrine or 20 mM caffeine. The present results suggest that citronellol lowers blood pressure by a direct effect on the vascular smooth muscle leading to vasodilation. PMID- 20002068 TI - The race toward 'ethically universally acceptable' human pluripotent (embryonic like) stem cells: only a problem of sources? AB - Over the past few years, several proposals aimed at procuring human pluripotent (embryonic-like) stem cells without involving the destruction of a human embryo have been proposed and widely discussed. This article focuses on a basic aspect of the debate, namely the plausibility of one or more of these new proposals being able to meet the ethical requirements that those who regard the human embryo as sacred have tried to impose on stem cells research in the last ten years. The thesis of the article is that focusing the discussion only on the sources of stem cells has prevented a full understanding of the foundation, meaning and scope of these ethical requirements. To substantiate this thesis, the article takes into consideration two issues: the first has to do with the potential of the cells obtained through some of the new approaches (iPS included), the second (and decisive) with the argument of the 'indirect complicity', applied to the use of 'contaminated' knowledge. PMID- 20002069 TI - Navigating the murky intersection between clinical and organizational ethics: a hybrid case taxonomy. AB - Ethical challenges that arise within healthcare delivery institutions are currently categorized as either clinical or organizational, based on the type of issue. Despite this common binary issue-based methodology, empirical study and increasing academic dialogue indicate that a clear line cannot easily be drawn between organizational and clinical ethics. Disagreement around end-of-life treatments, for example, often spawn value differences amongst parties at both organizational and clinical levels and requires a resolution to address both the case at hand and large-scale underlying system-level confounders. I refer to issues that contain elements of both clinical and organizational issues as hybrids and propose a new taxonomy to characterize hybrid cases. I contend that salient contextual features of an ethical issue, such as where it is identified, who it impacts and where it is ideally resolved in relation to its scope of impact, should inform procedure. Implementation of a Hybrid taxonomy viewing ethical issues as existing on a continuum furthers that end. The primary goals are to 1) systematize thinking about ethical issues that arise within healthcare delivery institutions and 2) allow the content of the ethical challenge to drive the process, rather than continuing to rely on the traditional binary issue-based choice. Failure to capture the complexity of hybrid situations perpetuates incomplete information and ultimately an inchoate resolution that creates more questions than answers. PMID- 20002070 TI - The ongoing charity of organ donation. Contemporary English Sunni fatwas on organ donation and blood transfusion. AB - BACKGROUND: Empirical studies in Muslim communities on organ donation and blood transfusion show that Muslim counsellors play an important role in the decision process. Despite the emerging importance of online English Sunni fatwas, these fatwas on organ donation and blood transfusion have hardly been studied, thus creating a gap in our knowledge of contemporary Islamic views on the subject. METHOD: We analysed 70 English Sunni e-fatwas and subjected them to an in-depth text analysis in order to reveal the key concepts in the Islamic ethical framework regarding organ donation and blood transfusion. RESULTS: All 70 fatwas allow for organ donation and blood transfusion. Autotransplantation is no problem at all if done for medical reasons. Allotransplantation, both from a living and a dead donor, appears to be possible though only in quite restricted ways. Xenotransplantation is less often mentioned but can be allowed in case of necessity. Transplantation in general is seen as an ongoing form of charity. Nearly half of the fatwas allowing blood transfusion do so without mentioning any restriction or problem whatsoever. The other half of the fatwas on transfusion contain the same conditional approval as found in the arguments pro organ transplantation. CONCLUSION: Our findings are very much in line with the international literature on the subject. We found two new elements: debates on the definition of the moment of death are hardly mentioned in the English Sunni fatwas and organ donation and blood transfusion are presented as an ongoing form of charity. PMID- 20002071 TI - Undignified bioethics. AB - The concept of dignity is pervasive in bioethics. However, some bioethicists have argued that it is useless on three grounds: that it is indeterminate; that it is reactionary; and that it is redundant. In response, a number of defences of dignity have recently emerged. All of these defences claim that when dignity is suitably clarified, it can be of great use in helping us tackle bioethical controversies. This paper rejects such defences of dignity. It outlines the four most plausible conceptions of dignity: dignity as virtuous behaviour; dignity as inherent moral worth; Kantian dignity; and dignity as species integrity. It argues that while each conception is coherent, each is also fundamentally flawed. As such, the paper argues for a bioethics without dignity: an 'undignified bioethics.' PMID- 20002072 TI - Surrogacy: donor conception regulation in Japan. AB - As of 2008, surrogacy is legal and openly practised in various places; Japan, however, has no regulations or laws regarding surrogacy. This paper reports the situation of surrogacy in Japan and in five other regions (the USA, the UK, Taiwan, Korea and France) to clarify the pros and cons of prohibiting surrogacy, along with the problems and issues relating to surrogacy compensation. Not only in a country such as France that completely prohibits surrogacy within the country, but also in a country such as the UK that allows non-commercial surrogacy, infertile couples travel overseas for the purpose of surrogacy. In addition, some couples might seek underground surrogacy if the government prohibits surrogacy. If an intended parent couple and a surrogate make an agreement among themselves and then a problem occurs, they cannot ask for support from professionals or bring a case to court, as can be observed in South Korea and Taiwan. We also conclude that there is little difference between commercial surrogacy and non-commercial surrogacy in the absence of a clear definition of 'reasonable expenses.' In the UK, the law does not allow surrogates to receive compensation. However, in reality, there may be little difference between the amounts paid to surrogates for profit in the US and those paid to surrogates for reasonable expenses in the UK. We conclude that the issue of surrogacy demands further discussion in Japan. PMID- 20002073 TI - Will biomedical enhancements undermine solidarity, responsibility, equality and autonomy? AB - Prominent thinkers such as Jurgen Habermas and Michael Sandel are warning that biomedical enhancements will undermine fundamental political values. Yet whether biomedical enhancements will undermine such values depends on how biomedical enhancements will function, how they will be administered and to whom. Since only few enhancements are obtainable, it is difficult to tell whether these predictions are sound. Nevertheless, such warnings are extremely valuable. As a society we must, at the very least, be aware of developments that could have harmful consequences. Indeed, if important values were to be jeopardized, we should take appropriate measures to protect them. This paper focuses on four central values: solidarity, personal responsibility, equality and autonomy. It delineates the conditions under which biomedical enhancements would undermine these values. It also details the circumstances under which these values would be unaffected by enhancements as well as those under which they would be promoted. Specifying these conditions is valuable; it would enable society to prepare appropriate ethical guidelines and policy responses in advance. PMID- 20002074 TI - Editorial for BJCP Special Obesity Edition. PMID- 20002075 TI - Pharmacotherapy for obesity. AB - Obesity is associated with increased risk of conditions such as hypertension, dyslipidaemia, diabetes mellitus, and obstructive sleep apnoea. Pharmacotherapy for obesity should be considered in combination with lifestyle changes in obese patients, or overweight patients with other conditions that put them at risk of developing heart disease. Sibutramine and orlistat are the only two anti-obesity medications approved for long-term use. Sibutramine is a serotonergic and adrenergic drug that reduces food intake. Orlistat is a gastrointestinal lipase inhibitor that interferes with fat absorption. However, it commonly causes flatulence and diarrhoea. Rimonabant is the first of a series of endocannabinoid receptor antagonists. It was approved by the Committee for Medicinal Products for Human Use of the European Medicines Agency (EMEA) as an adjunct to diet and exercise in treating obesity in 2006. However, despite the extensive clinical trial data, EMEA announced in 2008 that it has recommended suspension of rimonabant because of its psychiatric side effects. Studies evaluating the long term safety and efficacy of anti-obesity agents are needed. PMID- 20002076 TI - Recent progress in the genetics of common obesity. AB - The genetic contribution to interindividual variation in common obesity has been estimated at 40-70%. Yet, despite a relatively high heritability, the search for obesity susceptibility genes has been an arduous task. This paper reviews recent progress made in the obesity genetics field with an emphasis on established obesity susceptibility loci identified through candidate gene as well as genome wide studies. For the last 15 years, candidate gene and genome-wide linkage studies have been the two main genetic epidemiological approaches to identify genetic loci for common traits, yet progress has been slow and success limited. Only recently have candidate gene studies started to succeed; by means of large scale studies and meta-analyses at least five variants in four candidate genes have been found to be robustly associated with obesity-related traits. Genome wide linkage studies, however, have so far not been able to pinpoint genetic loci for common obesity. The genome-wide association approach, which has become available in recent years, has dramatically changed the pace of gene discoveries for common disease, including obesity. Three waves of large-scale high-density genome-wide association studies have already discovered at least 15 previously unanticipated genetic loci incontrovertibly associated with body mass index and extreme obesity risk. Although the combined contribution of these loci to the variation in obesity risk at the population level is small and their predictive value is typically low, these recently discovered loci are set to improve fundamentally our insights into the pathophysiology of obesity. PMID- 20002078 TI - Rise in antiobesity drug prescribing for children and adolescents in the UK: a population-based study. AB - WHAT IS ALREADY KNOWN ABOUT THIS SUBJECT: * The antiobesity drugs sibutramine and orlistat are not licensed for use in children and adolescents in the UK or USA. * Clinical trials suggest antiobesity drugs are effective and well-tolerated in obese adolescents. WHAT THIS STUDY ADDS: * Prescribing of unlicensed antiobesity drugs in children and adolescents has increased significantly in the past 8 years. * Most prescribed antiobesity drugs in children and adolescents are rapidly discontinued before patients can see clinical benefit, suggesting they are poorly tolerated or poorly efficacious. AIMS: The international childhood obesity epidemic has driven increased use of unlicensed antiobesity drugs, whose efficacy and safety are poorly studied in children and adolescents. We investigated the use of unlicensed antiobesity drugs (orlistat, sibutramine and rimonabant) in children and adolescents (0-18 years) in the UK. METHODS: Population-based prescribing data from the UK General Practice Research Database between 1 January 1999 and 31 December 2006. RESULTS: A total of 452 subjects received 1334 prescriptions during the study period. The annual prevalence of antiobesity drug prescriptions rose significantly from 0.006 per 1000 [95% confidence interval (CI) 0.0007, 0.0113] in 1999 to 0.091 per 1000 (95% CI 0.07, 0.11) in 2006, a 15-fold increase, with similar increases seen in both genders. The majority of prescriptions were made to those >or=14 years old, although 25 prescriptions were made for children <12 years old. Orlistat accounted for 78.4% of all prescriptions; only one patient was prescribed rimonabant. However, approximately 45% of the patients ceased orlistat and 25% ceased sibutramine after only 1 month. The estimated mean treatment durations for orlistat and sibutramine were 3 and 4 months, respectively. CONCLUSIONS: Prescribing of unlicensed antiobesity drugs in children and adolescents has dramatically increased in the past 8 years. The majority are rapidly discontinued before patients can see weight benefit, suggesting they are poorly tolerated or poorly efficacious when used in the general population. Further research into the effectiveness and safety of antiobesity drugs in clinical populations of children and adolescents is needed. PMID- 20002079 TI - New central targets for the treatment of obesity. AB - The review focuses on the central neuronal circuits involved in energy homeostasis and the opportunities these offer for pharmacological intervention to decrease feeding behaviour and reduce weight. This article is based on the presentation 'New central targets for the treatment of obesity' (Sargent, British Pharmacological society, Clinical Section Symposium, December 2008). Central neuronal substrates controlling weight offer numerous opportunities for pharmacological intervention. These opportunities range from non-specific enhancement of monoamine signalling (triple reuptake inhibitors) to targeting specific monoamine receptor subtypes (5-HT(2c) and 5-HT(6)). The data reviewed suggest that these approaches will lead to weight loss; whether this is sufficient to produce clinically meaningful effect remains to be determined. Combination therapy targeting more than one mechanism may be a means of increasing the magnitude of the response. Preclinical studies also suggest that novel approaches targeting specific neuronal pathways within the hypothalamus, e.g. MCH(1) receptor antagonism, offer an opportunity for weight reduction. However, these approaches are at an early stage and clinical studies will be needed to determine if these novel approaches lead to clinically meaningful weight loss and improvements in co-morbid conditions such as diabetes and cardiovascular disorders. PMID- 20002077 TI - Obesity treatment: novel peripheral targets. AB - Our knowledge of the complex mechanisms underlying energy homeostasis has expanded enormously in recent years. Food intake and body weight are tightly regulated by the hypothalamus, brainstem and reward circuits, on the basis both of cognitive inputs and of diverse humoral and neuronal signals of nutritional status. Several gut hormones, including cholecystokinin, glucagon-like peptide-1, peptide YY, oxyntomodulin, amylin, pancreatic polypeptide and ghrelin, have been shown to play an important role in regulating short-term food intake. These hormones therefore represent potential targets in the development of novel anti obesity drugs. This review focuses on the role of gut hormones in short- and long term regulation of food intake, and on the current state of development of gut hormone-based obesity therapies. PMID- 20002081 TI - Metformin increases plasma ghrelin in Type 2 diabetes. AB - WHAT IS ALREADY KNOWN ABOUT THIS SUBJECT: * Metformin, unlike the other major antihyperglycaemic drugs, is not associated with weight gain. * Ghrelin is an appetite-stimulating hormone whose concentrations vary in relation to food, obesity and diabetes control. * Reports are conflicting about how metformin affects ghrelin concentrations, and this study was aimed at resolving this issue in patients with Type 2 diabetes. WHAT THIS STUDY ADDS: * In this study an increase in ghrelin concentrations was seen in response to metformin treatment in patients with Type 2 diabetes. * This effect was opposite to what might be expected if the effect of metformin on weight control was mediated via suppression of ghrelin. * It is likely that the ghrelin response was secondary to improved glycaemic control. * Meal time changes in appetite and satiety did not correlate with changes in ghrelin, which suggests ghrelin may not be important in meal initiation. AIMS: Metformin treatment of Type 2 diabetes is not usually associated with weight gain, and may assist with weight reduction. Plasma ghrelin concentrations are inversely associated with obesity and food intake. Metformin might therefore affect ghrelin concentrations, although previous studies have shown variable results in this regard. The primary aim of this study was to determine the effect of metformin on plasma ghrelin, appetite and satiety in patients with Type 2 diabetes. METHODS: Eighteen patients with Type 2 diabetes were studied before and after 6 weeks of metformin treatment, which was titrated to 1 g b.d. On the study days patients were fed standard meals of 390 kcal at 08.00 and 12.30 h, plasma samples were collected at 15- and 30-min intervals, and appetite and satiety were measured on visual analogue scales. Changes in the area under the concentration-time curves (AUCs) of plasma ghrelin, insulin, glucose, appetite and satiety were assessed and examined for correlations with metformin AUCs. Changes in fasting adiponectin and leptin were also measured. RESULTS: Treatment with metformin increased the mean AUC (07.30-16.30 h) of plasma ghrelin by 24% (P= 0.003), while decreasing those of glucose by 19% (P < 0.001) and insulin by 19% (P= 0.001). No changes were detected in hunger and satiety, or in fasting adiponectin or leptin concentrations. There were no clear correlations between metformin plasma concentrations (AUC) and changes in plasma glucose, insulin or ghrelin. CONCLUSIONS: Treatment of Type 2 diabetes with metformin was associated with increased plasma ghrelin concentrations, without associated changes in hunger and satiety. PMID- 20002080 TI - Regulatory challenges for new drugs to treat obesity and comorbid metabolic disorders. AB - Obesity is a major cause of morbidity and mortality through cardio- and cerebrovascular diseases and cancer. The metabolic consequences of obesity include dyslipidaemia, hypertension, proinflammatory atherogenesis, pre-diabetes and Type 2 diabetes. For a significant proportion of patients, pharmacotherapy to tackle obesity is required as adjunctive support to diet, exercise and lifestyle modification. To this end, the pharmaceutical industry is pursuing many novel drug targets. Although this view is probably not justified, the recent tribulations of rimonabant have created a perception that the regulatory bar for the approval of antiobesity drugs has been raised. Although >5% of placebo subtracted weight loss maintained over 1 year is the primary efficacy end-point, it is improvements in cardiovascular risk factors that the Food and Drug Administration (FDA) and European Medicines Agency (EMEA) require to grant approval. Safety aspects are also critical in this indication. Many companies are now switching development of their antiobesity drug candidates into other metabolic disorders. Type 2 diabetes is accepted by the industry and FDA, but not EMEA, as the most appropriate alternative. On the other hand, improvements in plasma lipids produced by antiobesity drugs are moderate compared with established therapies, suggesting dyslipidaemia is not a viable development option. Metabolic Syndrome is not accepted by FDA or EMEA as a discrete disease and the agencies will not licence antiobesity drugs for its treatment. The regulatory environment for antiobesity drugs and the spectrum of indications for which they can be approved could change dramatically if positive data for sibutramine emerge from the SCOUT outcome trial. PMID- 20002082 TI - Pharmacokinetics and pharmacodynamics of LC15-0444, a novel dipeptidyl peptidase IV inhibitor, after multiple dosing in healthy volunteers. AB - WHAT IS ALREADY KNOWN ABOUT THIS SUBJECT: * The importance of efficient drug development using biomarkers has been increasingly emphasized, from preclinical studies to clinical trials. * However, as yet few validated or qualified biomarkers are used in early-stage drug development in terms of clinical pharmacology and disease pathophysiology. WHAT THIS STUDY ADDS: * This first-time in-human study provides evidence of the pharmacological activity of LC15-0444 in humans, by using dipeptidyl peptidase IV activity and active glucagon-like peptide-1 concentrations. * LC15-0444 possesses pharmacokinetic and pharmacodynamic characteristics that support a once-daily dosing regimen. AIMS: LC15-0444 is a selective and competitive inhibitor of dipeptidyl peptidase (DPP) IV with potential for the treatment of Type 2 diabetes. The aim was to investigate the pharmacokinetic (PK) and pharmacodynamic (PD) profiles after multiple oral ascending doses of LC15-0444 in healthy male subjects. METHODS: A dose block-randomized, double-blind, placebo-controlled, parallel group study was performed in three groups with 10 subjects (eight for active drug; two for placebo) per group; each group received 200, 400 or 600 mg of LC15-0444 once daily for 10 days. Blood and urine samples were collected up to 24 h after the first dosing and up to 72 h after the last dosing. RESULTS: The LC15-0444 concentration-time profiles exhibited characteristics of multicompartment disposition. No dose- or time-dependent change in PK parameters was observed. Mean elimination half-life was in a range 16.6-20.1 h in the dose groups. Mean renal clearance and fraction of unchanged drug excreted in urine was 18.6-21.9 and 0.40-0.48 l h(-1), respectively. In the steady state, mean accumulation ratios by dose groups were between 1.22 and 1.31. More than 80% inhibition of DPP IV activity from baseline was sustained for >24 h in all dose groups. CONCLUSIONS: This study provides evidence of the pharmacological activity of LC15 0444 in humans. LC15-0444 possesses PK and PD characteristics that support a once daily dosing regimen. PMID- 20002083 TI - Effect of efavirenz on high-density lipoprotein antioxidant properties in HIV infected patients. AB - WHAT IS ALREADY KNOWN ABOUT THIS SUBJECT: * In previous work, we showed a long term and concentration-dependent beneficial effect of the non-nucleoside reverse transcriptase inhibitor efavirenz (EFV) on high-density lipoproteins (HDL) in human immunodeficiency virus (HIV)-infected patients. * Furthermore, it has been suggested that instead of the current practice of only measuring HDL-chelesterol values, the evaluation of HDL function, namely its antioxidant properties, might be an improved tool for identifying subjects at increased risk for cardiovascular events. * Paraoxonase-1 (PON-1) is an enzyme associated with HDL that is responsible for HDL antioxidant function. WHAT THIS STUDY ADDS: * In the present work, we studied the effect of EFV on the activity of PON-1 and showed, for the first time, that EFV-based antiretroviral therapy is associated with a better antioxidant function, i.e. with a higher PON-1 activity. AIMS: A long-term and concentration-dependent beneficial effect of efavirenz (EFV) on cholesterol associated with high-density lipoprotein (HDL-c) in human immunodeficiency virus (HIV)-infected patients has been documented. Furthermore, it has been suggested that, instead of the current practice of only measuring HDL-c values, the evaluation of HDL quality might be an improved tool for identifying subjects at increased risk of cardiovascular events. Paraoxonase-1 (PON-1) is an enzyme associated with HDL that is involved in the onset of cardiovascular disease and responsible for HDL antioxidant function. The aim of the present study was to investigate the effect of EFV on the circulating activity of PON-1 in HIV infected patients. METHODS: The patients included were adults with a documented HIV-1 infection, nontreated or treated with antiretroviral regimens including EFV 600 mg once daily as first therapeutic regimen for at least 3 months. The influence of treatment with EFV, HDL-c and CD4 cell count on PON-1 activity was analysed. RESULTS: HIV-infected White patients treated with EFV had higher PON-1 activity [77.35 U l(-1) (65.66, 89.04)] (P < 0.05) and higher PON-1 activity : HDL-c ratio [1.88 (1.49, 2.28)] (P < 0.01) than untreated patients. PON-1 activity was higher in Black patients (P < 0.001) and in patients with a CD4 cell count >500 cells ml(-1) (P= 0.0120). CONCLUSIONS: EFV-based antiretroviral regimens are associated with HDL particles with a better antioxidant function, i.e. with a higher PON-1 activity. The PON-1 activity of Black patients is higher than that found in Whites regardless of treatment. Ethnicity should be taken into consideration when studying drug effects on PON-1 activity. PMID- 20002084 TI - Effect of renal impairment on the pharmacokinetics of the GLP-1 analogue liraglutide. AB - WHAT IS ALREADY KNOWN ABOUT THIS SUBJECT: * Patients with Type 2 diabetes are likely to have or to develop renal impairment, which affects the pharmacokinetics of some antidiabetic treatments. * Whether dosing of the once-daily human glucagon-like peptide-1 analogue liraglutide should be modified in patients with renal impairment has not previously been studied. WHAT THIS STUDY ADDS: * Renal dysfunction was not found to increase the exposure of liraglutide. * Hence, no dose adjustment is expected to be required in patients with Type 2 diabetes and renal impairment treated with liraglutide. AIMS: To investigate whether dose adjustment of the once-daily human glucagon-like peptide-1 analogue liraglutide is required in patients with varying stages of renal impairment. METHODS: A cohort of 30 subjects, of whom 24 had varying degrees of renal impairment and six had normal renal function, were given a single dose of liraglutide, 0.75 mg subcutaneously, and completed serial blood sampling for plasma liraglutide measurements for pharmacokinetic estimation. RESULTS: No clear trend for change in pharmacokinetics was evident across groups with increasing renal dysfunction. While the between-group comparisons of the area under the liraglutide concentration-curve (AUC) did not demonstrate equivalence [estimated ratio AUC(severe)/AUC(healthy) 0.73, 90% confidence interval (CI) 0.57, 0.94; and AUC (continuous ambulatory peritoneal dialysis)(CAPD)/AUC(healthy) 0.74, 90% CI 0.56, 0.97], the regression analysis of log(AUC) for subjects with normal renal function and mild-to-severe renal impairment showed no significant effect of decreasing creatinine clearance on the pharmacokinetics of liraglutide. The expected AUC ratio between the two subjects with the lowest and highest creatinine clearance in the study was estimated to be 0.88 (95% CI 0.58, 1.34) (NS). Degree of renal impairment did not appear to be associated with an increased risk of adverse events. CONCLUSIONS: This study indicated no safety concerns regarding use of liraglutide in patients with renal impairment. Renal dysfunction was not found to increase exposure of liraglutide, and patients with Type 2 diabetes and renal impairment should use standard treatment regimens of liraglutide. There is, however, currently limited experience with liraglutide in patients beyond mild-stage renal disease. PMID- 20002085 TI - Pharmacokinetics, metabolism and bioavailability of the triazole antifungal agent voriconazole in relation to CYP2C19 genotype. AB - WHAT IS ALREADY KNOWN ABOUT THIS SUBJECT: * Pharmacokinetic variability of voriconazole is largely caused by CYP3A4- and CYP2C19-mediated metabolism. * Oral bioavailability of voriconazole has been claimed to be almost 100%, thus facilitating a change from intravenous to oral application without dose adjustment. WHAT THIS STUDY ADDS: * For the first time voriconazole exposure after intravenous and oral administration in relation to CYP2C19 activity is reported. * In addition, the predominant metabolic pathway is the hydroxylation that seems to be influenced by the CYP2C19 genotype. * Enterohepatic circulation of both hydroxylated metabolites must be anticipated. AIMS: The aim was to determine the pharmacokinetics of voriconazole after a single oral dose in comparison with intravenous (i.v.) administration in healthy individuals stratified according to the cytochrome P450 (CYP) 2C19 genotype. In addition, the possible metabolic pathways and their modulation according to CYP2C19 genotype were investigated after oral and i.v. administration of voriconazole. METHODS: In a single-centre, open-label, two-period crossover study 20 participants received single doses of 400 mg voriconazole orally and 400 mg voriconazole intravenously in randomized order. Blood and urine samples were collected up to 96 h post dose and the voriconazole and three major metabolites were quantified by high performance liquid chromatography coupled to mass spectroscopy. RESULTS: Absolute oral bioavailability of voriconazole was 82.6% (74.1, 91.0). It ranged from 94.4% (78.8, 109.9) in CYP2C19 poor metabolizers to 75.2% (62.9, 87.4) in extensive metabolizers. In contrast to voriconazole and its N-oxide, the plasma concentrations of both hydroxylated metabolites showed a large second peak after 24 h. Independent of the route of administration, voriconazole partial metabolic hydroxylation after i.v. administration was eightfold higher compared with N oxidation [48.8 ml min(-1) (30.5, 67.1) vs. 6.1 ml min(-1) (4.1, 8.0)]. The formation of the metabolites was related to CYP2C19 activity. CONCLUSIONS: Independent of the route of administration, voriconazole exposure was three times higher in CYP2C19 poor metabolizers compared with extensive metabolizers. Voriconazole has a high bioavailability with no large differences between the CYP2C19 genotypes. The hydroxylation pathway of voriconazole elimination exceeded the N-oxidation, both influenced by the CYP2C19 genotype. PMID- 20002086 TI - Relationship between blood alcohol concentration on admission and outcome in dimethoate organophosphorus self-poisoning. AB - WHAT IS ALREADY KNOWN ABOUT THIS SUBJECT: * Acute alcohol intoxication often complicates acute organophosphorus pesticide poisoning. * No data are available on how alcohol intoxication affects outcome in acute organophosphorus pesticide poisoning. * In particular, the relationships between plasma alcohol concentration and plasma organophosphorus concentration or outcome are unclear. WHAT THIS STUDY ADDS: * Alcohol co-ingestion is associated with higher concentrations of the organophosphorus insecticide dimethoate, probably due to larger ingestions. * The higher concentrations of dimethoate found with alcohol co-ingestion increase the risk of death in dimethoate poisoning. There was no detectable effect of the alcohol itself on outcome. * Efforts to reduce deaths from insecticide self-poisoning may benefit from concurrent efforts to reduce alcohol consumption. AIMS: Many patients acutely poisoned with organophosphorus insecticides have co-ingested alcohol. Although clinical experience suggests that this makes management more difficult, the relationship between plasma concentration of alcohol and insecticide is unknown. We aimed to determine whether acute intoxication results in ingestion of larger quantities of insecticide in dimethoate self-poisoning and a worse clinical outcome. METHODS: We set up a prospective study of acute dimethoate self-poisoning in Sri Lankan district hospitals. An admission plasma sample was analysed to identify the ingested insecticide; in patients with detectable dimethoate, plasma alcohol was measured. RESULTS: Plasma from 37 of 72 (51.4%) dimethoate-poisoned patients had detectable alcohol [median concentration 1.10 g l(-1)[110 mg dl( 1)][interquartile range (IQR) 0.78-1.65]] a median of 3 h post ingestion. The median plasma dimethoate concentration was higher in patients who had ingested alcohol [479 micromol l(-1) (IQR 268-701) vs. 145 micromol l(-1) (IQR 25-337); P < 0.001]. Plasma dimethoate concentration was positively correlated with plasma alcohol (Spearman's rho= 0.34; P= 0.0032). The median alcohol concentration was higher in the 21 patients who died compared with survivors (0.94 vs. 0.0 g l(-1), P= 0.018). Risk of death was greater amongst individuals who consumed alcohol [odds ratio (OR) 4.3, 95% confidence interval (CI) 1.2, 16.4]; this risk was abolished by controlling for dimethoate concentration (OR 0.3, 95% CI 0.0, 8.8), indicating that deaths were not due to the direct toxic effects of alcohol. CONCLUSIONS: Alcohol co-ingestion is associated with higher plasma concentrations of dimethoate and increased risk of death. Larger studies are required to assess this finding's generalizability, since efforts to reduce deaths from self poisoning may benefit from concurrent efforts to reduce alcohol consumption. PMID- 20002087 TI - Inhibition of oral midazolam clearance by boosting doses of ritonavir, and by 4,4 dimethyl-benziso-(2H)-selenazine (ALT-2074), an experimental catalytic mimic of glutathione oxidase. AB - WHAT IS ALREADY KNOWN ABOUT THIS SUBJECT: * The viral protease inhibitor ritonavir is known to inhibit clearance of intravenous midazolam. * ALT-2074, a catalytic mimic of glutathione oxidase, inhibits human cytochrome P450 3A (CYP3A) isoforms in vitro. WHAT THIS STUDY ADDS: * Short-term administration of low-dose ritonavir increases area under the plasma concentration curve following oral midazolam by a factor of 28. * Therefore ritonavir is an appropriate positive control inhibitor for clinical drug interaction studies involving CYP3A substrates. * Midazolam clearance is weakly inhibited by ALT-2074, consistent with its in vitro profile. AIMS: We evaluated whether 'boosting' doses of ritonavir can serve as a positive control inhibitor for pharmacokinetic drug-drug interaction studies involving cytochrome P450 3A (CYP3A). The study also determined whether 4,4-dimethyl-benziso-(2H)-selenazine (ALT-2074), an investigational organoselenium compound that acts as a catalytic mimic of glutathione oxidase, inhibits CYP3A metabolism in vivo. METHODS: Thirteen healthy volunteers received single 3-mg oral doses of midazolam on three occasions: in the control condition, during co-treatment with low-dose ritonavir (three oral doses of 100 mg over 24 h), and during co-treatment with ALT-2074 (three oral doses of 80 mg over 24 h). RESULTS: Ritonavir increased mean (+/-SE) total area under the curve (AUC) for midazolam by a factor of 28.4 +/- 4.2 (P < 0.001), and reduced oral clearance to 4.2 +/- 0.5% of control (P < 0.001). In contrast, ALT 2074 increased midazolam AUC by 1.25 +/- 0.11 (P < 0.05), and reduced oral clearance to 88 +/- 8% of control. CONCLUSIONS: Low-dose ritonavir produces extensive CYP3A inhibition exceeding that of ketoconazole (typically 10- to 15 fold midazolam AUC enhancement), and is a suitable positive control index inhibitor for drug-drug interaction studies. ALT-2074 inhibits CYP3A metabolism to a small degree that is of uncertain clinical importance. PMID- 20002088 TI - Pharmacokinetic assessment of a five-probe cocktail for CYPs 1A2, 2C9, 2C19, 2D6 and 3A. AB - WHAT IS ALREADY KNOWN ABOUT THIS SUBJECT: * Numerous cocktails using concurrent administration of several cytochrome P450 (CYP) isoform-selective probe drugs have been reported to investigate drug-drug interactions in vivo. * This approach has several advantages: characterize the inhibitory or induction potential of compounds in development toward the CYP enzymes identified in vitro in an in vivo situation, assess several enzymes in the same trial, and have complete in vivo information about potential CYP-based drug interactions. WHAT THIS STUDY ADDS: * This study describes a new cocktail containing five probe drugs that has never been published. * This cocktail can be used to test the effects of a new chemical entity on multiple CYP isoforms in a single clinical study: CYP1A2 (caffeine), CYP2C9 (warfarin), CYP2C19 (omeprazole), CYP2D6 (metoprolol), and CYP3A (midazolam) and was designed to overcome potential liabilities of other reported cocktails. AIMS: To assess the pharmacokinetics (PK) of selective substrates of CYP1A2 (caffeine), CYP2C9 (S-warfarin), CYP2C19 (omeprazole), CYP2D6 (metoprolol) and CYP3A (midazolam) when administered orally and concurrently as a cocktail relative to the drugs administered alone. METHODS: This was an open-label, single dose, randomized, six-treatment six-period six-sequence William's design study with a wash-out of 7 or 14 days. Thirty healthy male subjects received 100 mg caffeine, 100 mg metoprolol, 0.03 mg kg(-1) midazolam, 20 mg omeprazole and 10 mg warfarin individually and in combination (cocktail). Poor metabolizers of CYP2C9, 2C19 and 2D6 were excluded. Plasma samples were obtained up to 48 h for caffeine, metoprolol and omeprazole, 12 h for midazolam, 312 h for warfarin and the cocktail. Three different validated liquid chromatography tandem mass spectrometry methods were used. Noncompartmental PK parameters were calculated. Log-transformed C(max), AUC(last) and AUC for each analyte were analysed with a linear mixed effects model with fixed term for treatment, sequence and period, and random term for subject within sequence. Point estimates (90% CI) for treatment ratios (individual/cocktail) were computed for each analyte C(max), AUC(last) and AUC. RESULTS: There was no PK interaction between the probe drugs when administered in combination as a cocktail, relative to the probes administered alone, as the 90% CI of the PK parameters was within the prespecified bioequivalence limits of 0.80, 1.25. CONCLUSION: The lack of interaction between probes indicates that this cocktail could be used to evaluate the potential for multiple drug-drug interactions in vivo. PMID- 20002089 TI - Potentially inappropriate prescribing in an Irish elderly population in primary care. AB - WHAT IS ALREADY KNOWN ABOUT THIS SUBJECT: * Potentially inappropriate prescribing in older people is a well-documented problem and has been associated with adverse drug reactions and hospitalization. * Beers' criteria, Screening Tool of Older Persons' potentially inappropriate Prescriptions (STOPP) and Screening Tool to Alert doctors to Right Treatment (START) are screening tools that have been formulated to help physicians and pharmacists identify potentially inappropriate prescribing and potential prescribing omissions. * The prevalence of potentially inappropriate prescribing and prescribing omissions in the elderly population presenting to hospital with acute illness is high according to STOPP and START criteria. WHAT THIS STUDY ADDS: * Potential errors of prescribing and of omission of medicines are prevalent among medically stable older people in primary care. * Screening tools should be incorporated into the everyday practice of primary care doctors and community pharmacists as a means of preventing potential errors of prescribing commission and prescribing omission in older people. AIMS: Screening tools have been formulated to identify potentially inappropriate prescribing (IP) in older people. Beers' criteria are the most widely used but have disadvantages when used in Europe. New IP screening tools called Screening Tool of Older Person's Prescriptions (STOPP) and Screening Tool to Alert doctors to Right Treatment (START) have been developed to identify potential IP and potential prescribing omissions (PPOs). The aim was to measure the prevalence rates of potential IP and PPOs in primary care using Beers' criteria, STOPP and START. METHODS: Case records of 1329 patients >or=65 years old from three general practices in one region of southern Ireland were studied. The mean age +/- SD of the patients was 74.9 +/- 6.4 years, 60.9% were female. Patients' current diagnoses and prescription medicines were reviewed and the Beers' criteria, STOPP and START tools applied. RESULTS: The total number of medicines prescribed was 6684; median number of medicines per patient was five (range 1-19). Overall, Beers' criteria identified 286 potentially inappropriate prescriptions in 18.3% (243) of patients, whilst the corresponding IP rate identified by STOPP was 21.4% (284), in respect of 346 potentially inappropriate prescriptions. A total of 333 PPOs were identified in 22.7% (302) of patients using the START tool. CONCLUSION: Potentially inappropriate drug prescribing and errors of drug omission are highly prevalent among older people living in the community. Prevention strategies should involve primary care doctors and community pharmacists. PMID- 20002090 TI - Pharmacokinetic and clinical profile of a novel formulation of bosentan in children with pulmonary arterial hypertension: the FUTURE-1 study. AB - WHAT IS ALREADY KNOWN ABOUT THIS SUBJECT: * Exposure to bosentan was lower in paediatric pulmonary arterial hypertension (PAH) patients treated with the marketed adult formulation at a dose of about 2 mg kg(-1) when compared with adult PAH patients. * In healthy adult subjects, bosentan pharmacokinetics are less than dose-proportional at doses of >or=500 mg. WHAT THIS STUDY ADDS: * The pharmacokinetics of a new paediatric bosentan formulation were characterized in paediatric PAH patients. * The level of exposure to bosentan as observed in adult PAH patients cannot be reached in paediatric patients with b.i.d. dosing. * In paediatric PAH patients, nondose-proportional pharmacokinetics of bosentan occur at lower doses when compared with healthy adult subjects. AIM: To show equivalent bosentan exposure in paediatric patients with pulmonary arterial hypertension (PAH) when compared with a cohort of historical controls of adult PAH patients using a newly developed paediatric formulation. METHODS: Thirty-six paediatric PAH patients were enrolled in this multicentre, prospective, open-label, noncontrolled study and treated for 4 weeks with bosentan 2 mg kg(-1) b.i.d. and then for 8 weeks with 4 mg kg(-1) b.i.d. Blood samples were taken for pharmacokinetic purposes. Exploratory efficacy measurements included World Health Organization (WHO) functional class and parent's and clinician's Global Clinical Impression scales. RESULTS: Comparing children with a historical group of adults, the geometric mean ratio (90% confidence interval) of the area under the plasma concentration-time curve was 0.54 (0.37, 0.78), i.e. children had lower exposure to bosentan than adults. Bosentan concentrations following doses of 2 and 4 mg kg(-1) were similar. Improvements in WHO functional class and the Global Clinical Impression scales occurred mainly in bosentan-naive patients, whereas the rare worsenings occurred in patients already on bosentan prior to study initiation. The paediatric formulation was well accepted and bosentan well tolerated in this study. No cases of elevated liver enzymes or anaemia were reported. CONCLUSIONS: Exposure to bosentan, as shown comparing the results from this study with those from a study in adults, was different in paediatric and adult PAH patients. Since FUTURE-1 and past studies suggest a favourable benefit-risk profile for bosentan at 2 mg kg(-1) b.i.d., this dose is recommended for children with PAH. The new paediatric formulation was well tolerated. PMID- 20002091 TI - Exposure to folic acid antagonists during the first trimester of pregnancy and the risk of major malformations. AB - WHAT IS ALREADY KNOWN ABOUT THIS SUBJECT: * Previous studies have suggested a tendency of antifolate drugs to be associated with higher rates of neural tube defects. WHAT THIS STUDY ADDS: * This study makes use of the data on abortuses, which is missed in many other studies. In this case, the abortion data were critical. * The study documents that clinicians should avoid, as much as possible, the use of folic acid antagonists during the first trimester of pregnancy, when embryogenesis takes place. AIM: To investigate the safety of folic acid antagonists during the first trimester of pregnancy in a large cohort. METHODS: Computerized databases for medications dispensed from 1998 to 2007 to women registered in 'Clalit' HMO, Israel southern district, was linked with maternal and infant hospitalization records, and to therapeutics abortions data. The risk for adverse pregnancy outcomes of folic acid antagonists exposure was assessed by adjusting for known confounders. RESULTS: Eighty-four thousand, eight hundred and twenty-three infants were born and 998 therapeutic abortions took place; 571 fetuses and infants were exposed to one or more folic acid antagonists in the first trimester of pregnancy. Exposure was associated with an overall increased risk of congenital malformations [odds ratio (OR) 2.43, 95% confidence interval (CI) 1.92, 3.08], due mainly to increased risk for neural tube (adjusted OR 6.5, 95% CI 4.34, 9.15) and cardiovascular defects (OR 1.76, CI 1.05, 2.95). CONCLUSION: First-trimester exposure to folic acid antagonists is associated with increased risk of congenital malformations. PMID- 20002092 TI - Self report may lead to underestimation of 'wrong dose' medication errors. PMID- 20002095 TI - Role of the signal peptide in the synthesis and processing of the glucagon-like peptide-1 receptor. AB - BACKGROUND AND PURPOSE: The glucagon-like peptide-1 receptor (GLP-1R) belongs to Family B of the G protein-coupled receptor superfamily and is a target for treatment of type 2 diabetes. Family B G protein-coupled receptors contain a putative N-terminal signal peptide, but its role in receptor synthesis and trafficking are unclear. Further, the signal peptide is not cleaved in at least one family member. EXPERIMENTAL APPROACH: We examined receptor glycosylation and the role of the signal peptide in GLP-1R synthesis and trafficking using constructs containing epitope tags at the N- and/or C-terminus and in which the signal peptide sequence was either present or absent. KEY RESULTS: The signal peptide was absolutely required for GLP-1R synthesis but could be substituted to some extent by increasing positive charge in the N-terminal region of the receptor flanking the signal peptide. The signal peptide is cleaved during synthesis and processing of the receptor. An enhanced GFP-epitope tag at the N terminus of the receptor permitted synthesis of the receptor but blocked signal peptide cleavage and prevented trafficking to the plasma membrane. Cleavage site mutation allowed synthesis of a full-length receptor, blocked signal peptide cleavage and caused retention within the endoplasmic reticulum. CONCLUSIONS AND IMPLICATIONS: Signal peptide cleavage was not essential for receptor synthesis but was obligatory for processing and trafficking of receptors to the plasma membrane. Further, the GLP-1R is subject to N-linked glycosylation and only the mature, fully glycosylated form of the receptor is present in the plasma membrane. Inhibition of glycosylation prevents processing and cell surface expression of the GLP-1R. PMID- 20002096 TI - Differential actions of urocortins on neurons of the myenteric division of the enteric nervous system in guinea pig distal colon. AB - BACKGROUND AND PURPOSE: Urocortins (Ucns) 1, 2 and 3 are corticotropin-releasing factor (CRF)-related neuropeptides and may be involved in neural regulation of colonic motor functions. Nevertheless, details of the neural mechanism of action for Ucns have been unclear. We have, here, tested the hypothesis that Ucns act in the enteric nervous system (ENS) to influence colonic motor behaviour. EXPERIMENTAL APPROACH: We used intracellular recording with 'sharp' microelectrodes, followed by intraneuronal injection of biocytin, and immunohistochemical localization of CRF(1) and CRF(2) receptors in guinea pig colonic tissue. KEY RESULTS: Application of Ucn1 depolarized membrane potentials and elevated excitability in 58% of AH-type and 60% of S-type colonic myenteric neurons. In most of the neurons tested, depolarizing responses evoked by Ucn-1 were suppressed by the CRF(1) receptor antagonist NBI 27914, but were unaffected by the CRF(2) receptor antagonist antisauvagine-30. The selective CRF(2) receptor agonists, Ucn2 and Ucn3, evoked depolarizing responses in 12 and 8% of the AH type myenteric neurons, respectively, and had no effect on S-type neurons. Antisauvagine-30, but not NBI 27914, suppressed these Ucn2- and Ucn3-evoked responses. Immunohistochemical staining identified CRF(1) as the predominant CRF receptor subtype expressed by ganglion cell somas, while CRF(2)-immunoreactive neuronal somas were sparse. Ucns did not affect excitatory synaptic transmission in the ENS. CONCLUSIONS AND IMPLICATIONS: The results suggest that Ucns act as neuromodulators to influence myenteric neuronal excitability. The excitatory action of Ucn1 in myenteric neurons was primarily at CRF(1) receptors, and the excitatory action of Ucn2 and Ucn3 was at CRF(2) receptors. PMID- 20002098 TI - Positron emission tomography imaging of tissue P-glycoprotein activity during pregnancy in the non-human primate. AB - BACKGROUND AND PURPOSE: Changes in tissue P-glycoprotein (P-gp) activity during pregnancy could affect the pharmacokinetics and thus the efficacy and toxicity of many drugs. Therefore, using positron emission tomography (PET) imaging, we tested whether gestational age affects tissue P-gp activity in the pregnant non human primate, Macaca nemestrina. EXPERIMENTAL APPROACH: Mid-gestational (day 75 +/- 13, n= 7) and late-gestational (day 150 +/- 10, n= 5) age macaques were imaged after administration of a prototypic P-gp substrate, (11)C-verapamil (13.7 75.4 MBq.kg(-1)), before and during intravenous infusion of a P-gp inhibitor, cyclosporin A (CsA) (12 or 24 mg.kg(-1).h(-1)). Accumulation of radioactivity in the fetal liver served as a reporter of placental P-gp activity. P-gp activity was expressed as CsA-induced percent change in the ratio of the area (0-9 min) under the (11)C-radioactivity concentration-time curve in the tissue (AUC(tissue)) to that in the maternal plasma (AUC(plasma)). KEY RESULTS: The CsA induced change in AUC(fetal liver)/AUC(maternal)(plasma) of (11)C-radioactivity significantly increased from mid- (35 +/- 25%) to late gestation (125 +/- 66%). Likewise, the CsA-induced change in AUC(maternal brain)/AUC(plasma) increased from mid- (172 +/- 80%) to late gestation (337 +/- 148%). The AUC ratio for the other maternal tissues was not significantly affected. Neither the CsA blood concentrations nor the level of circulating (11)C-verapamil metabolites were significantly affected by gestational age. CONCLUSIONS AND IMPLICATIONS: P-gp activity at the blood-brain barrier and the placental barrier in the macaque increased with gestational age. If replicated in humans, the exposure of the fetus and maternal brain to P-gp substrate drugs, and therefore their efficacy and toxicity, will change during pregnancy. PMID- 20002097 TI - Procalcitonin in sepsis and systemic inflammation: a harmful biomarker and a therapeutic target. AB - The worldwide yearly mortality from sepsis is substantial, greater than that of cancer of the lung and breast combined. Moreover, its incidence is increasing, and its response to therapy has not appreciably improved. In this condition, the secretion of procalcitonin (ProCT), the prohormone of calcitonin, is augmented greatly, attaining levels up to thousands of fold of normal. This hypersecretion emanates from multiple tissues throughout the body that are not traditionally viewed as being endocrine. The serum values of ProCT correlate with the severity of sepsis; they recede with its improvement and worsen with exacerbation. Accordingly, as highlighted in this review, serum ProCT has become useful as a biomarker to assist in the diagnosis of sepsis, as well as related infectious or inflammatory conditions. It is also a useful monitor of the clinical course and prognosis, and sensitive and specific assays have been developed for its measurement. Moreover, it has been demonstrated that the administration of ProCT to septic animals greatly increases mortality, and several toxic effects of ProCT have been elucidated by in vitro experimental studies. Antibodies have been developed that neutralize the harmful effects of ProCT, and their use markedly decreases the symptomatology and mortality of animals that harbour a highly virulent sepsis analogous to that occurring in humans. This therapy is facilitated by the long duration of serum ProCT elevation, which allows for a broad window of therapeutic opportunity. An experimental groundwork has been established that suggests a potential applicability of such therapy in septic humans. PMID- 20002099 TI - Colitis affects the smooth muscle and neural response to motilin in the rabbit antrum. AB - BACKGROUND AND PURPOSE: The underlying mechanisms of gastric dysfunction during or after an episode of intestinal inflammation are poorly understood. This study investigated the effects of colitis on the contractile effects of motilin, an important endocrine regulator of gastric motility, in the antrum. EXPERIMENTAL APPROACH: Myeloperoxidase (MPO) activity, NF-kappaB activity and motilin receptor density were determined in the antrum of rabbits 5 days after the induction of 2,4,6-trinitrobenzenesulphonic acid colitis. Smooth muscle and neural responses to motilin were studied in antral smooth muscle strips in vitro. KEY RESULTS: Colitis did not affect MPO activity, but increased NF-kappaB activity in the antrum. Motilin receptor density in the antrum was not affected. Under control conditions, motilin induced a slowly developing tonic smooth muscle contraction. Five days post-inflammation, tonic contractions to motilin were reduced and preceded by a rapid initial contraction. Other kinases were recruited for the phosphorylation of myosin light chain (MLC) (a multi-functional MLC kinase), and for the inhibition of MLC phosphatase (Rho kinase in addition to protein kinase C) to mediate the motilin-induced contractions during inflammation. Colitis potentiated the cholinergic neural on-contractions in the antrum. This was associated with a hyper-reactivity to motilin and an increased muscle response to ACh. CONCLUSIONS AND IMPLICATIONS: Colitis altered the course of the motilin induced smooth muscle contraction in the antrum. This involved changes in the kinases phosphorylating MLC. Increased cholinergic excitability to motilin in the antrum may play a role in the pathogenesis of inflammation-associated gastric motility disorders. PMID- 20002100 TI - Arachidonic acid release mediated by OX1 orexin receptors. AB - BACKGROUND AND PURPOSE: We have previously shown that lipid mediators, produced by phospholipase D and C, are generated in OX(1) orexin receptor signalling with high potency, and presumably mediate some of the physiological responses to orexin. In this study, we investigated whether the ubiquitous phospholipase A(2) (PLA(2)) signalling system is also involved in orexin receptor signalling. EXPERIMENTAL APPROACH: Recombinant Chinese hamster ovary-K1 cells, expressing human OX(1) receptors, were used as a model system. Arachidonic acid (AA) release was measured from (3)H-AA-labelled cells. Ca(2+) signalling was assessed using single-cell imaging. KEY RESULTS: Orexins strongly stimulated [(3)H]-AA release (maximally 4.4-fold). Orexin-A was somewhat more potent than orexin-B (pEC(50) = 8.90 and 8.38 respectively). The concentration-response curves appeared biphasic. The release was fully inhibited by the potent cPLA(2) and iPLA(2) inhibitor, methyl arachidonyl fluorophosphonate, whereas the iPLA(2) inhibitors, R- and S bromoenol lactone, caused only a partial inhibition. The response was also fully dependent on Ca(2+) influx, and the inhibitor studies suggested involvement of the receptor-operated influx pathway. The receptor-operated pathway, on the other hand, was partially dependent on PLA(2) activity. The extracellular signal regulated kinase, but not protein kinase C, were involved in the PLA(2) activation at low orexin concentrations. CONCLUSIONS AND IMPLICATIONS: Activation of OX(1) orexin receptors induced a strong, high-potency AA release, possibly via multiple PLA(2) species, and this response may be important for the receptor operated Ca(2+) influx. The response coincided with other high-potency lipid messenger responses, and may interact with these signals. PMID- 20002101 TI - TREK-1 channels do not mediate nitrergic neurotransmission in circular smooth muscle from the lower oesophageal sphincter. AB - BACKGROUND AND PURPOSE: The ionic mechanisms underlying nitrergic inhibitory junction potentials (IJPs) in gut smooth muscle remain a matter of debate. Recently, it has been reported that opening of TWIK-related K(+) channel 1 (TREK 1) K(+) channels contributes to the nitrergic IJP in colonic smooth muscle. We investigated the effects of TREK-1 channel blockers on nitrergic neurotransmission in mouse and opossum lower oesophageal sphincter (LOS) circular smooth muscle (CSM). EXPERIMENTAL APPROACH: The effects of TREK-1 channel blockers were characterized pharmacologically in murine and opossum gut smooth muscle using conventional intracellular and tension recordings. KEY RESULTS: In LOS, L-methionine depolarized the resting membrane potential (RMP) but did not inhibit the nitrergic IJP. Cumulative application of theophylline hyperpolarized the RMP and inhibited the nitrergic IJP concentration dependently. The induced membrane hyperpolarization was prevented by pre-application of caffeine, but not by 1H-[1,2,4]oxadiazolo-[4,3-a]quinoxalin-1-one. 8-Br-cAMP significantly hyperpolarized membrane potential and increased the amplitude of the nitrergic IJP. In opossum LOS muscle strips, L-methionine increased resting tone but had no effect on nerve-mediated LOS relaxation. On the other hand, theophylline markedly inhibited tone. In CSM from mouse proximal colon, L-methionine caused modest inhibition of nitrergic IJPs. CONCLUSIONS AND IMPLICATIONS: TREK-1 channels were not involved in the nitrergic IJP in LOS CSM. Not only does L-methionine have no effect on the nitrergic IJP or LOS relaxation, but the effect of theophylline appears to be due to interruption of Ca(2+)-releasing pathways (i.e. caffeine like effect) rather than via blockade of TREK-1 channels. PMID- 20002102 TI - Antidepressant-like effects of cannabidiol in mice: possible involvement of 5 HT1A receptors. AB - BACKGROUND AND PURPOSE: Cannabidiol (CBD) is a non-psychotomimetic compound from Cannabis sativa that induces anxiolytic- and antipsychotic-like effects in animal models. Effects of CBD may be mediated by the activation of 5-HT(1A) receptors. As 5-HT(1A) receptor activation may induce antidepressant-like effects, the aim of this work was to test the hypothesis that CBD would have antidepressant-like activity in mice as assessed by the forced swimming test. We also investigated if these responses depended on the activation of 5-HT(1A) receptors and on hippocampal expression of brain-derived neurotrophic factor (BDNF). EXPERIMENTAL APPROACH: Male Swiss mice were given (i.p.) CBD (3, 10, 30, 100 mg*kg(-1)), imipramine (30 mg*kg(-1)) or vehicle and were submitted to the forced swimming test or to an open field arena, 30 min later. An additional group received WAY100635 (0.1 mg*kg(-1), i.p.), a 5-HT(1A) receptor antagonist, before CBD (30 mg*kg(-1)) and assessment by the forced swimming test. BDNF protein levels were measured in the hippocampus of another group of mice treated with CBD (30 mg*kg( 1)) and submitted to the forced swimming test. KEY RESULTS: CBD (30 mg*kg(-1)) treatment reduced immobility time in the forced swimming test, as did the prototype antidepressant imipramine, without changing exploratory behaviour in the open field arena. WAY100635 pretreatment blocked CBD-induced effect in the forced swimming test. CBD (30 mg*kg(-1)) treatment did not change hippocampal BDNF levels. CONCLUSION AND IMPLICATIONS: CBD induces antidepressant-like effects comparable to those of imipramine. These effects of CBD were probably mediated by activation of 5-HT(1A) receptors. PMID- 20002103 TI - Transient supersensitivity to alpha-adrenoceptor agonists, and distinct hyper reactivity to vasopressin and angiotensin II after denervation of rat tail artery. AB - BACKGROUND AND PURPOSE: Vascular 'denervation' hyper-reactivity has generally been investigated 1-2 weeks after administration of chemicals that temporarily prevent transmitter release, but do not necessarily inactivate the neuronal noradrenaline transporters (NETs). We have investigated the reactivity of rat tail arteries over longer periods after removing the terminals by surgical denervation. EXPERIMENTAL APPROACH: Two and 7 weeks after denervation, myography was used to assess contractions of isolated arterial segments to phenylephrine, methoxamine, clonidine, vasopressin and angiotensin II (AII). Denervation was confirmed by lack of tyrosine hydroxylase immunoreactive nerve terminals. KEY RESULTS: The NET inhibitor, desmethylimipramine, increased the pEC(50) for phenylephrine in control, but not denervated arteries after both 2 and 7 weeks. Relative to controls, pEC(50)s for phenylephrine (with desmethylimipramine), methoxamine, clonidine and vasopressin were increased at 2 but not 7 weeks after denervation. The pEC(50) for phenylephrine in the absence of desmethylimipramine was greater than control after both 2 and 7 weeks' denervation. The maximum contraction to vasopressin was larger than in controls at 2 but not 7 weeks after denervation, whereas contractions to AII were markedly enhanced at both time points. CONCLUSIONS AND IMPLICATIONS: Increased vascular reactivity to alpha(1)- and alpha(2)-adrenoceptor agonists, and vasopressin is transient following denervation. After 7 weeks, increased reactivity to phenylephrine can be entirely accounted for by the loss of NETs. Maintained supersensitivity to AII indicates that denervation differentially and selectively affects vascular reactivity to circulating vasoconstrictor agents. This might explain persistent vasoconstriction in denervated skin of humans after nerve injuries. PMID- 20002104 TI - Evidence that the plant cannabinoid cannabigerol is a highly potent alpha2 adrenoceptor agonist and moderately potent 5HT1A receptor antagonist. AB - BACKGROUND AND PURPOSE: Cannabis is the source of at least seventy phytocannabinoids. The pharmacology of most of these has been little investigated, three notable exceptions being Delta(9)-tetrahydrocannabinol, cannabidiol and Delta(9)-tetrahydrocannabivarin. This investigation addressed the question of whether the little-studied phytocannabinoid, cannabigerol, can activate or block any G protein-coupled receptor. EXPERIMENTAL APPROACH: The [(35)S]GTPgammaS binding assay, performed with mouse brain membranes, was used to test the ability of cannabigerol to produce G protein-coupled receptor activation or blockade. Its ability to displace [(3)H]CP55940 from mouse CB(1) and human CB(2) cannabinoid receptors and to inhibit electrically evoked contractions of the mouse isolated vas deferens was also investigated. KEY RESULTS: In the brain membrane experiments, cannabigerol behaved as a potent alpha(2)-adrenoceptor agonist (EC(50)= 0.2 nM) and antagonized the 5-HT(1A) receptor agonist, R-(+)-8 hydroxy-2-(di-n-propylamino)tetralin (apparent K(B)= 51.9 nM). At 10 microM, it also behaved as a CB(1) receptor competitive antagonist. Additionally, cannabigerol inhibited evoked contractions of the vas deferens in a manner that appeared to be alpha(2)-adrenoceptor-mediated (EC(50)= 72.8 nM) and displayed significant affinity for mouse CB(1) and human CB(2) receptors. CONCLUSIONS AND IMPLICATIONS: This investigation has provided the first evidence that cannabigerol can activate alpha(2)-adrenoceptors, bind to cannabinoid CB(1) and CB(2) receptors and block CB(1) and 5-HT(1A) receptors. It will now be important to investigate why cannabigerol produced signs of agonism more potently in the [(35)S]GTPgammaS binding assay than in the vas deferens and also whether it can inhibit noradrenaline uptake in this isolated tissue and in the brain. PMID- 20002105 TI - Alagebrium attenuates acute methylglyoxal-induced glucose intolerance in Sprague Dawley rats. AB - BACKGROUND AND PURPOSE: Alagebrium is a breaker of cross-links in advanced glycation endproducts. However, the acute effects of alagebrium on methylglyoxal (MG), a major precursor of advanced glycation endproducts have not been reported. MG is a highly reactive endogenous metabolite, and its levels are elevated in diabetic patients. We investigated whether alagebrium attenuated the acute effects of exogenous MG on plasma MG levels, glucose tolerance and distribution of administered MG in different organs in Sprague-Dawley rats. EXPERIMENTAL APPROACH: We measured MG levels (by HPLC), glucose tolerance, adipose tissue glucose uptake, GLUT4, insulin receptor and insulin receptor substrate 1 (IRS-1) protein expression, and phosporylated IRS-1 in rats treated with MG at doses of either 17.25 mg*kg(-1) i.p. (MG-17 i.p.) or 50 mg*kg(-1) i.v. (MG-50 i.v.) with or without alagebrium, 100 mg*kg(-1) i.p. KEY RESULTS: Alagebrium attenuated the increased MG levels in the plasma, aorta, heart, kidney, liver, lung and urine after MG administration. In MG-treated rats, glucose tolerance was impaired, plasma insulin levels were higher and insulin-stimulated glucose uptake by adipose tissue was reduced, relative to the corresponding control groups. In rats treated with MG-50 i.v., GLUT4 protein expression and IRS-1 tyrosine phosphorylation were decreased. Alagebrium pretreatment attenuated these effects of MG. In an in vitro assay, alagebrium reduced the amount of detectable MG. CONCLUSIONS AND IMPLICATIONS: Alagebrium acutely attenuated MG-induced glucose intolerance, suggesting a possible preventive role for alagebrium against the harmful effects of MG. PMID- 20002106 TI - Factors associated with mucoid transition of Pseudomonas aeruginosa in cystic fibrosis patients. AB - Although the mucoid form of Pseudomonas aeruginosa (Pa) is largely responsible for the progression of lung disease in cystic fibrosis (CF), the relationship between factors relating daily-care regimes to mucoidy acquisition are as yet poorly investigated. Fifty-two CF patients registered at the CF centre of Dijon, France, were retrospectively evaluated from the date of Pa colonization either to the first positive sputum culture for mucoid Pa (n = 26) or to the last culture in which the Pa remained non-mucoid (n = 26). All clinical, pathological and therapeutic events were recorded. The association between the parameters collected and mucoid transition of Pa was assessed in a Cox model with time dependant covariables. The mean follow-up was 4.7 + or - 4.3 years. Three independent parameters were associated with the higher risk of mucoid transition of Pa: persistence of Pa in sputum (OR 7.89; p <0.01), use of inhaled bronchodilators (OR 3.40; p = 0.04), and the use of inhaled colimycin (OR 4.04; p = 0.02). Isolation of Staphylococcus aureus, Haemophilus influenzae or Streptococcus pneumoniae in sputum was associated with a lower risk (OR 0.24; p < 0.01). Mucoid transition of Pa was associated with variables that reflected the severity of both lung disease and Pa colonization. Although they do not lead to prophylactic measures, these results corroborate the need to avoid Pa persistence. PMID- 20002107 TI - Activity of the new cephalosporin CXA-101 (FR264205) against Pseudomonas aeruginosa isolates from chronically-infected cystic fibrosis patients. AB - Chronic respiratory infection caused by Pseudomonas aeruginosa is the main driver of morbidity and mortality in cystic fibrosis (CF) patients. The development of resistance to all available antibiotics is a frequent outcome of these infections. The present study aimed to evaluate the activity of the new cephalosporin CXA-101 (FR264205) against a collection of 100 isolates obtained from 50 CF patients from two Spanish hospitals. The collection included the first (early) and the last (late) available isolate from each patient (average interval 68 +/- 39 months). The MIC50 and MIC90 of CXA-101 were 0.5 and 2 mg/L and the geometric mean MIC was 0.7 mg/L; the MICs for 95% of the isolates were <=8 mg/L (tentative breakpoint). Only meropenem yielded comparable results, although the MIC90 of this antibiotic was significantly higher (8 mg/L). CXA-101 showed conserved activity against a high proportion of isolates resistant to each of the antibiotics tested (ceftazidime, cefepime, piperacillin-tazobactam, imipenem, meropenem, levofloxacin and tobramycin), with MIC50 values of 1-2 mg/L. Moreover, CXA-101 retained good activity against multidrug-resistant strains, with MIC50 and MIC90 values of 2 and 16 mg/L. CXA-101 was also active against late CF isolates (the MIC for 96% was <=8 mg/L); it was the only antibiotic tested to which a similar percentage of early and late isolates was susceptible. These results show that, despite a slight increase in MICs, major cross-resistance to CXA-101 did not develop during treatment of CF patients with the currently available antipseudomonal agents. Therefore, CXA-101 is envisaged as a valuable alternative for the treatment of chronic respiratory infection caused by P. aeruginosa in CF patients. PMID- 20002108 TI - Modified sequential multiplex PCR for determining capsular serotypes of invasive pneumococci recovered from Seville. AB - The heptavalent pneumococcal vaccine's introduction resulted in a decline in invasive disease caused by Streptococcus pneumoniae, but was accompanied by an increase in non-vaccine serotypes. We evaluated a modified scheme of the sequential multiplex PCRs adapted to the prevalence of serotypes in Seville (Spain) for determining capsular serotypes of S. pneumoniae invasive clinical isolates. In adults, the modified scheme allowed us to type 73% with the first three reactions, and 92% with two additional PCRs. In paediatric patients, it allowed us to type 73.5% with the first three reactions, and 90% with the two additional PCRs. The multiplex PCR approach was successfully adapted to target the serotypes most prevalent in Seville. PMID- 20002110 TI - Statistical recommendations for papers submitted to Developmental Medicine & Child Neurology. AB - The use of statistics in medical diagnoses and biomedical research may affect whether an individual may live or die, whether their health is protected or jeopardized. Because society depends on sound statistical practice, all practitioners of statistics, whatever their training or occupation, have social obligations to perform their work in a professional, competent, and ethical manner. PMID- 20002112 TI - Evidence of impaired neurocognitive functioning in school-age children awaiting cardiac surgery. AB - AIM: Children with congenital heart disease (CHD) are at risk of developing neurocognitive problems. However, as these problems are usually identified after cardiac surgery, it is unclear whether they resulted from the surgery or whether they pre-existed and hence might be explained by complications and events associated with the heart disease itself. The purpose of this study was to examine whether neurocognitive deficits commonly reported after cardiac surgery are present before surgery. METHOD: Forty-five children (22 males, 23 females; mean age 11 y 6 mo, SD 3 y 0 mo) with cyanotic and acyanotic heart diseases scheduled for elective cardiac surgery were compared with 41 healthy peers (17 males, 24 females; mean age 11 y 10 mo, SD 2 y 10 mo) for attention and processing speed, construction, motor speed, motor planning and fluency, and visual memory. Twenty-three children in the patient group were awaiting their first cardiac surgery and 22 were awaiting follow-up surgery. RESULTS: The patients showed manifest neurocognitive difficulties. Their performance was inferior to that of the healthy comparison group for motor planning (p=0.02) and visual memory (p=0.01). The same neurocognitive profile was found in the group of patients awaiting their first cardiac operation. INTERPRETATION: School-age children with various forms of CHD are at risk of neurocognitive impairments before cardiac surgery. PMID- 20002113 TI - Free testosterone levels in umbilical-cord blood predict infant head circumference in females. AB - AIM: Fetal androgens influence fetal growth as well as postnatal neurocognitive ability. However, to our knowledge, no published study has prospectively examined the impact of early-life androgens on infant brain growth. We report the association between circulating fetal androgen levels, measured from umbilical cord blood at birth, and a proxy measure of brain growth: head circumference. METHOD: Participants were 82 unselected female infants from a large representative birth cohort (mean gestational age 39.4 wks, SD 1.7). Umbilical cord blood was obtained at birth and analysed for androgen concentrations (total testosterone, androstenedione, dehyrdroepiandrosterone, and its sulphated metabolite). Head circumference and two other measures of growth - weight (mean 3311.4 g, SD 461.3) and length - were measured within 3 days of birth and again at approximately 1 year of age (mean age 13.1 mo, SD 1.1). RESULTS: Multivariate linear regressions found an inverse association between levels of free testosterone and growth in head circumference (correlation=-.24), even when adjusting for sociodemographic/obstetric covariates and head size at birth. Growth in weight and length could not be predicted by free testosterone concentration. INTERPRETATION: This is the first report of an association between prenatal androgen levels and postnatal growth in head circumference. These findings suggest that early-life androgens may impact brain development during infancy. PMID- 20002114 TI - Prevalence of motor-skill impairment in preterm children who do not develop cerebral palsy: a systematic review. AB - AIM: Motor skill impairment is a common negative outcome in children born preterm who do not develop cerebral palsy (CP). This study aimed to conduct a systematic review of current data to provide an accurate estimate of the prevalence of non CP motor impairment in preterm children at school age. METHOD: We searched the Medline, PubMed, and PsycInfo databases and relevant journals to identify all studies published post-1990 that reported the prevalence of motor impairment in school-aged children born preterm (<37 wks' gestation) using standardised motor assessment batteries. We applied a range of exclusionary criteria, with 11 studies included in the final analyses. We identified two levels of motor impairment commonly reported - mild-moderate and moderate - and conducted a random effects meta-analysis to produce a prevalence estimate for each. RESULTS: The pooled estimate for mild-moderate impairment in preterm children was 40.5/100. and for moderate motor impairment the estimate was 19.0/100. There was also a trend for lower motor impairment levels in samples born before 1990 compared with those born after 1990. INTERPRETATION: Children born preterm are at increased risk of motor impairment, with prevalence three to four times greater than in the general population. This highlights the need for improved surveillance and intervention strategies in this group of children. PMID- 20002115 TI - Visuospatial attention disturbance in Duchenne muscular dystrophy. AB - AIM: The cognitive deficits present in the Duchenne muscular dystrophy (DMD) are not yet well characterized. Attention, considered to be the brain mechanism responsible for the selection of sensory stimuli, could be disturbed in DMD, contributing, at least partially, to the observed global cognitive deficit. The aim of this study was to investigate attentional function in individuals with DMD. METHOD: Twenty-five males (mean age 12y; SD 2y 2mo) with DMD and 25 healthy males (mean age 12y; SD 2y) were tested in a visuospatial task (Posner computerized test). They were instructed to respond as quickly as possible to a lateralized visual target stimulus with the ipsilateral hand. Their attention was automatically orientated by a peripheral prime stimulus or, alternatively, voluntarily orientated by a central spatially informative cue. RESULTS: The main result obtained was that the attentional effect (sum of the benefit and the cost of attention) did not differ between the two groups in the case of automatic attention (p=0.846) but was much larger for individuals with DMD than for comparison individuals in the case of voluntary attention (p<0.001). INTERPRETATION: The large voluntary attentional effect exhibited by the participants with DMD seems similar to that of younger children, suggesting that the disease is associated with delayed maturation of voluntary attention mechanisms. PMID- 20002116 TI - The effect of a basic home stimulation programme on the development of young children infected with HIV. AB - AIMS: The human immunodeficiency virus (HIV) potentially causes a significant encephalopathy and resultant developmental delay in infected children. The aim of this study was to determine whether a home-based intervention programme could have an impact on the neurodevelopmental status of children infected with HIV. METHOD: A longitudinal, randomized, controlled trial was conducted. A total of 122 children aged less than 2 years 6 months were assigned to either a comparison or an experimental group. Children in the experimental group were given a home stimulation programme that was updated every 3 months. The home programme included activities to promote motor, cognitive, and speech and language development. Children in the comparison group received no developmental intervention. Children were assessed by a blinded assessor at baseline, 6 months, and 12 months using the Bayley Scales of Infant Development, 2nd edition. RESULTS: The children in this study came from poor socioeconomic backgrounds and their nutritional status was suboptimal. The experimental group included 60 children (30 males, 30 females) with a mean age of 18 months (SD 8.1 mo). The comparison group included 62 children (32 males, 30 females) with a mean age 19 months (SD 8.2 mo). Cognitive and motor development were severely affected at baseline, with 52% of the children having severe cognitive delay and 72% having severe motor delay at baseline. Children in the experimental group showed significantly greater improvement in cognitive (p=0.010) and motor (p=0.020) development over time than children in the comparison group. INTERPRETATION: A home stimulation programme taught to the caregiver can significantly improve cognitive and motor development in young children infected with HIV. PMID- 20002117 TI - Attentional and executive impairments in children with spastic cerebral palsy. AB - AIM: Children with cerebral palsy (CP) are reported to have learning and social problems. The aim of the present study was to examine whether children with CP have impairments in attention or executive function. METHOD: We examined attention and executive function with standardized neuropsychological measures in a group of children with unilateral (n=15) or bilateral (n=18) spastic CP (14 females, 19 males, mean age 11y 4mo, SD 1y 1mo, range 9y 1mo-13y 7mo; Gross Motor Function Classification System level I n=22, II n=3, III n=6, and IV n=2). Performance was compared with test norms. RESULTS: Verbal cognitive functioning fell within the normal range, whereas sustained (p=0.001) and divided attention (p<0.001) were found to be impaired. Greater impairment was observed in executive function in general (p<0.001) and in inhibition (p=0.038) and shifting (p<0.001) in particular. No significant difference was found between types of CP (unilateral and bilateral). Performance of all timed tasks was slower than the test norm (p<0.00). INTERPRETATION: The finding of slower performances across tasks may indicate a general impairment in efficiency of information processing in relation to white-matter lesions. Impairments in attention and executive functions are present in children with CP and may help to explain why these children have increased social and learning problems. PMID- 20002118 TI - Predictive value of definitely abnormal general movements in the general population. AB - AIM: Definitely abnormal general movements in populations of high-risk infants predict serious neurodevelopmental impairment. This study aimed to assess predictive values of definitely abnormal general movements at 3 months for serious neurodevelopmental impairment in a representative sample of the general population. METHOD: A prospective cohort study of 455 3-month-old infants was performed (241 females, 214 males; mean birthweight 3452g, SD 604g; mean gestational age 39.4wks, SD 1.96; n=32 born preterm). At enrolment, general movement quality was assessed by means of video recording. At 4 years, all participants were reassessed by well-baby health clinicians; if serious neurodevelopmental impairment was identified, clinical records were reviewed. Predictive values of definitely abnormal general movement quality for major neurodevelopmental impairment were calculated. RESULTS: Five children were diagnosed as having a major neurodevelopmental disorder with serious implications for daily life, including three children with cerebral palsy (CP). Three out of the five had shown definitely abnormal general movements; they had lesions involving the periventricular white matter. Two children did not show definitely abnormal general movements; one had unilateral spastic CP due to a cortical lesion and the other had ataxia due to cerebellar atrophy. The positive predictive value of definitely abnormal general movements for major neurodevelopmental impairment was 18/100, and for CP it was 12/100. Negative predictive values approached 100%. INTERPRETATION: The good predictive value of general movement assessment in high-risk populations cannot be generalized to the general population. PMID- 20002120 TI - 'Non-accidental brain injury: mechanisms and imponderables'. PMID- 20002122 TI - Characterization of motor control in handwriting difficulties in children with or without developmental coordination disorder. AB - AIM: The purpose of this study was to characterize handwriting deficits in children with developmental coordination disorder (DCD) using computerized movement analyses. METHOD: Seventy-two children (40 females, 32 males; mean age 7 y, SD 7 mo; range 6 y 2 mo to 7 y 11 mo) with handwriting deficits (33 with DCD, 39 without DCD); and 22 age- and sex-matched children without handwriting deficits were asked to perform handwriting tasks on a digital tablet for the collection of kinematic and kinetic data. Practice times required to achieve automation of movement when writing an unfamiliar character were used to assess the motor learning of handwriting. The children were asked to copy three simple and three complex characters, and the velocity and axial pen force used for corresponding strokes were compared. RESULTS: The attainment of automated handwriting was markedly slower in children with handwriting deficits and DCD, who used a faster stroke velocity to write simple characters (1.22 times those without handwriting deficits), but when writing complex characters, their stroke velocity and pen force were lower (0.85 and 0.89 times those without handwriting deficits, respectively). INTERPRETATION: By linking the results with neuromotor control theories, it was determined that children with DCD have difficulties performing the open-loop and closed-loop movements required for fluent handwriting. PMID- 20002125 TI - Pyruvate dehydrogenase complex deficiency: four neurological phenotypes with differing pathogenesis. AB - AIM: To describe the phenotype and genotype of pyruvate dehydrogenase complex (PDHc) deficiency. METHOD: Twenty-two participants with enzymologically and genetically confirmed PDHc deficiency were analysed for clinical and imaging features over a 15-year period. RESULTS: Four groups were identified: (1) those with neonatal encephalopathy with lactic acidosis (one male, four females; diagnosis at birth); (2) those with non-progressive infantile encephalopathy (three males, three females; age at diagnosis 2-9mo); (3) those with Leigh syndrome (eight males; age at diagnosis 1-13mo); and (4) those with relapsing ataxia (three males; 18-30mo). Seventeen mutations involved PDHA1 (a hotspot was identified in exons 6, 7, and 8 in seven males with Leigh syndrome or recurrent ataxia). Mutations in the PDHX gene (five cases) were correlated with non progressive encephalopathy and long-term survival in four cases. INTERPRETATION: Two types of neurological involvement were identified. Abnormal prenatal brain development resulted in severe non-progressive encephalopathy with callosal agenesis, gyration anomalies, microcephaly with intrauterine growth retardation, or dysmorphia in both males and females (12 cases). Acute energy failure in infant life produced basal ganglia lesions with paroxysmal dystonia, neuropathic ataxia due to axonal transport dysfunction, or epilepsy only in males (11 cases). The ketogenic diet improved only paroxysmal dysfunction, providing an additional argument in favour of paroxysmal energy failure. PMID- 20002126 TI - Relationship between gross motor capacity and daily-life mobility in children with cerebral palsy. AB - AIM: The aim of this study was to examine the relationship between gross motor capacity and daily-life mobility in children with cerebral palsy (CP) and to explore the moderation of this relationship by the severity of CP. METHOD: Cross sectional analysis in a cohort study with a clinic-based sample of children with CP (n=116; 76 males, 40 females; mean age 6 y 3 mo, SD 12 mo, range 4 y 8 mo-7 y 7 mo) was performed. Gross motor capacity was assessed by the Gross Motor Function Measure (GMFM-66). Daily-life mobility was assessed using the Pediatric Evaluation of Disability Inventory (PEDI): Functional Skills Scale (FSS mobility) and Caregiver Assistance Scale (CAS mobility). Severity of CP was classified by the Gross Motor Function Classification System (48% level I, 17% level II, 15% level III, 8% level IV, 12% level V), type of motor impairment (85% spastic, 12% dyskinetic, 3% ataxic), and limb distribution (36% unilateral, 49% bilateral spastic). RESULTS: Scores on the GMFM-66 explained 90% and 84% respectively, of the variance of scores on PEDI-FSS mobility and PEDI-CAS mobility. Limb distribution moderated the relationship between scores on the GMFM-66 and the PEDI-FSS mobility, revealing a weaker relationship in children with unilateral spastic CP (24% explained variance) than in children with bilateral spastic CP (91% explained variance). INTERPRETATION: In children aged 4 to 7 years with unilateral spastic CP, dissociation between gross motor capacity and daily-life mobility can be observed, just as in typically developing peers. PMID- 20002127 TI - Leisure activity preferences for 6- to 12-year-old children with cerebral palsy. AB - AIM: The objective was to describe leisure activity preferences of children with cerebral palsy (CP) and their relationship to participation. Factors associated with greater interest in leisure activities were identified. METHOD: Fifty-five school-aged children (36 males, 19 females; mean age 9 y 11 mo; range 6 y 1 mo-12 y 11 mo) with CP (Gross Motor Function Classification System [GMFCS]) level I 62%, level II 22%, level III-IV 16%; 33.3% hemiplegia, 29.6% diplegia, 25.9% quadriplegia, 11.2% other) who could complete the Preferences for Activities of Children (PAC) were recruited. RESULTS: Social and recreational activities were most preferred, and self-improvement activities were least preferred. Younger age, higher motivation, and IQ predicted interest in active-physical activities (r(2)=0.39). Negative reaction to failure was associated with less preference for social activities (r(2)=0.16), whereas increased prosocial behaviours were related to greater preference for recreational (r(2)=0.13) and self-improvement activities; the latter is also predicted by older age (r(2)=0.24). Interest in skill-based activities was greater in females and in children who were highly motivated, younger, and had greater motor limitations (r(2)=0.51). The findings suggest that personal factors and functional abilities influence leisure activity preferences. High preference for certain activities was not always associated with involvement in these activities. INTERPRETATION: Determination of preferences is inherent to child-centred practice and should, therefore, be part of the evaluation process. Rehabilitation strategies can minimize barriers to leisure participation, such as fear of failure, low motivation, or environmental obstacles. PMID- 20002128 TI - Predicting school readiness from neurodevelopmental assessments at age 2 years after respiratory distress syndrome in infants born preterm. AB - AIM: To determine whether neurodevelopmental outcomes at the age of 2 years accurately predict school readiness in children who survived respiratory distress syndrome after preterm birth. METHOD: Our cohort included 121 preterm infants who received surfactant and ventilation and were enrolled in a randomized controlled study of inhaled nitric oxide for respiratory distress syndrome. Abnormal outcomes at the age of 2 years were defined as neurosensory disability (cerebral palsy, blindness, or bilateral hearing loss) or delay (no neurosensory disability but Bayley Scales of Infant Development mental or performance developmental index scores <70). School readiness (assessed at a mean age of 5y 6mo, SD 1y) was determined using neurodevelopmental assessments of motor, sensory, receptive vocabulary, perceptual, conceptual, and adaptive skills. RESULTS: The mean birthweight of the cohort (57 males, 64 females) was 987g (SD 374), and the mean gestational age was 27.3 weeks (SD 2.6). At the age of 2 years, the neurodevelopmental classification was 'disabled' in 11% and 'delayed' in 23%. At the age of 5 years 6 months, intensive special education was required for 11% and some special education for 21%. Disability and delay at the age of 2 years were 92% and 50% predictive of lack of school readiness respectively, whereas only 15% of children who were normal at the age of 2 years were not school ready at the later assessment. Children with delay at 2 years were more likely to need special education if they were socially disadvantaged. INTERPRETATION: Without preschool developmental supports, preterm survivors living in poverty will require more special education services. PMID- 20002129 TI - A new case of creatine transporter deficiency associated with mild clinical phenotype and a novel mutation in the SLC6A8 gene. PMID- 20002130 TI - Apolipoprotein E genotype and cerebral palsy. AB - AIM: Apolipoprotein E (APOE, protein; [ApoE, gene]) is a lipid transport protein abundantly present in brain cells. We investigated whether the APOE genotype is associated with cerebral palsy (CP) and whether patients with CP with comorbid conditions and more severe neurological deficits are likely to have a particular genotype. METHOD: In a cross-sectional study, 243 individuals with spastic CP (135 males, 108 females; mean age at data collection 11 year ([SD 6y 7mo], 34% with hemiplegia, 37% with diplegia, 29% with triplegia/tetraplegia; 44% with mild motor involvement), 31% with moderate motor involvement, 25% with severe motor involvement, were compared with healthy individuals matched by age, race, and sex to analyse the association between APOE genotype and the incidence of CP. Associations between the APOE genotype and the incidence of comorbidities and neurological deficits were studied in the group with CP. RESULTS: The APOE epsilon2epsilon3 genotype was significantly more prevalent in the group with CP (11%) than the comparison group (5%) (odds ratio [OR] 2.8; 95% confidence interval [CI] 1.01-7.66). The presence of the epsilon2 allele raised the probability of having CP (OR 3.2; 95% CI 1.27-8.27). The presence of ApoE epsilon4 was not significantly different among groups. No relation was found between APOE genotype and severity of neurological deficit or distribution of motor involvement. Four patients with CP presented the epsilon4epsilon4 genotype, and all exhibited epilepsy and microcephaly. Eleven of 12 individuals with CP and macrocephaly carried the epsilon3epsilon3 genotype. INTERPRETATION: A higher prevalence of the APOE epsilon2 genotype was found among those with CP. The association of microcephaly and epilepsy with the epsilon4epsilon4 genotype and the association of macrocephaly with epsilon3 demand further investigation. PMID- 20002131 TI - 'Relabelling the preserved speech variant of Rett syndrome?'. PMID- 20002132 TI - Genetic analyses of Pseudomonas aeruginosa isolated from healthy captive snakes: evidence of high inter- and intrasite dissemination and occurrence of antibiotic resistance genes. AB - Faecal carriage of Pseudomonas aeruginosa was investigated by selective plating and PCR identification test, among healthy captive snakes from zoological and private collections from France as well as from wild snakes from Guinea. P. aeruginosa faecal carriage among captive snakes was high (72 out of 83 individuals), but low among wild specimen (3 out of 23 individuals). Genetic diversity analyses of the isolates, based on SpeI-PFGE profiles, evidenced five dominant clones or clonal complexes spreading among snakes within a site and between sites and persisting over time. Similar clones or clonal complexes were detected from mouth swabs of the owners and from water and preys used to feed the snakes, evidencing various sources of snake colonization and the first cases of P. aeruginosa cross-contamination between snakes and owners. These observations led to the conclusion that P. aeruginosa behaves as an opportunistic species within snakes in captivity and that colonization and dissemination occurs consecutively to processes similar to those identified within the hospital. Antibiotic susceptibility testing showed that most isolates had a wild-type resistance profile except for one persistent clone isolated from both snakes and preys that harboured multiple antimicrobial resistance genes mediated by an integron carrying the qacH, aadB, aadA2 and cmlA10 cassettes, and a tetA(C) carrying transposon. Biocides or antibiotics used in the zoological garden could have led to the acquisition of this integron. PMID- 20002133 TI - Abundances of crenarchaeal amoA genes and transcripts in the Pacific Ocean. AB - Planktonic Crenarchaea are thought to play a key role in chemolithotrophic ammonia oxidation, a critical step of the marine nitrogen (N) cycle. In this study, we examined the spatial distributions of ammonia-oxidizing Crenarchaea across a large (approximately 5200 km) region of the central Pacific Ocean. Examination of crenarchaeal 16S rRNA, ammonia monooxygenase subunit A (amoA) genes, and amoA transcript abundances provided insight into their spatial distributions and activities. Crenarchaeal gene abundances increased three to four orders of magnitude with depth between the upper ocean waters and dimly lit waters of the mesopelagic zone. The resulting median value of the crenarchaeal amoA: 16S rRNA gene ratio was 1.3, suggesting the majority of Crenarchaea in the epi- and mesopelagic regions of the Pacific Ocean have the metabolic machinery for ammonia oxidation. Crenarchaeal amoA transcript abundances typically increased one to two orders of magnitude in the transitional zone separating the epipelagic waters from the mesopelagic (100-200 m), before decreasing into the interior of the mesopelagic zone. The resulting gene copy normalized transcript abundances revealed elevated amoA expression in the upper ocean waters (0-100 m) where crenarchaeal abundances were low, with transcripts decreasing into the mesopelagic zone as crenarchaeal gene abundances increased. These results suggest ammonia-oxidizing Crenarchaea are active contributors to the N cycle throughout the epi- and mesopelagic waters of the Pacific Ocean. PMID- 20002134 TI - Anthropogenic disturbance affects the structure of bacterial communities. AB - Patterns of taxa abundance distributions are the result of the combined effects of historical and biological processes and as such are central to ecology. It is accepted that a taxa abundance distribution for a given community of animals or plants following a perturbation will typically change in structure from one of high evenness to increasing dominance. Subsequently, such changes in evenness have been used as indicators of biological integrity and environmental assessment. Here, using replicated experimental treehole microcosms perturbed with different concentrations of the pollutant pentachlorophenol, we investigated whether changes in bacterial community structure would reflect the effects of anthropogenic stress in a similar manner to larger organisms. Community structure was visualized using rank-abundance plots fitted with linear regression models. The slopes of the regression models were used as a descriptive statistic of changes in evenness over time. Our findings showed that bacterial community structure reflected the impact and the recovery from an anthropogenic disturbance. In addition, the intensity of impact and the rate of recovery to pre perturbation structure were dose-dependent. These properties of bacterial community structures may potentially provide a metric for environmental assessment and regulation. PMID- 20002135 TI - Fusarium spp. associated with rice Bakanae: ecology, genetic diversity, pathogenicity and toxigenicity. AB - African and Asian populations of Fusarium spp. (Gibberella fujikuroi species complex) associated with Bakanae of rice (Oryzae sativa L.) were isolated from seeds and characterized with respect to ecology, phylogenetics, pathogenicity and mycotoxin production. Independent of the origin, Fusarium spp. were detected in the different rice seed samples with infection rate ranges that varied from 0.25% to 9%. Four Fusaria (F. andiyazi, F. fujikuroi, F. proliferatum and F. verticillioides) were found associated with Bakanae of rice. While three of the Fusaria were found in both African and Asian seed samples, F. fujikuroi was only detected in seed samples from Asia. Phylogenetic studies showed a broad genetic variation among the strains that were distributed into four different genetic clades. Pathogenicity tests showed that all strains reduced seed germination and possessed varying ability to cause symptoms of Bakanae on rice, some species (i.e. F. fujikuroi) being more pathogenic than others. The ability to produce fumonisins (FB(1) and FB(2)) and gibberellin A3 in vitro also differed according to the Fusarium species. While fumonisins were produced by most of the strains of F. verticillioides and F. proliferatum, gibberellin A3 was only produced by F. fujikuroi. Neither fumonisin nor gibberellin was synthesized by most of the strains of F. andiyazi. These findings provide new information on the variation within the G. fujikuroi species complex associated with rice seed and Bakanae disease. PMID- 20002136 TI - Top-down control of microbial activity and biomass in an Arctic soil ecosystem. AB - Globally, soil microbes preside over vast carbon stores, and both microbial biomass and activity are known to be regulated by bottom-up controls, that is, limitation by nutrients and energy. However, there is evidence that grazing by protozoans exerts top-down controls on biomass. Here, we investigate top-down control by phage on soil microbes using an experimental site near Barrow, Alaska (71 degrees N, 157 degrees W) during the 2007 growing season. Soil measurements were taken from sites that covered a range of microtopographical features within a drained and thawed lake basin including high- and low-centred ice-wedge polygons to estimate the availability of carbon and nitrogen for microbes. Using both field and laboratory experiments, we successfully increased both microbial biomass and respiration by decreasing phage populations. The addition of carbon and nutrients to soils had no significant effects on biomass or respiration, indicating a lack of bottom-up controls. Additionally, we present the first use of tea extracts as a potent anti-phage agent in soils. Our results suggest that top-down controls, such as phage predation, are critical to regulation of microbial activities in Arctic soils. PMID- 20002137 TI - Seasonal variations of phage life strategies and bacterial physiological states in three northern temperate lakes. AB - The current consensus concerning the prevalence of lytic and lysogenic phage life cycles in aquatic systems is that the host physiological state may influence viral strategies, lysogeny being favoured when hosts have reduced metabolic rates. We explored this hypothesis, by following phage cycle dynamics, host physiological state and metabolic activity over an annual cycle in three lakes subjected to strong seasonal fluctuations, including 4-5 months of ice cover. We observed marked seasonal dynamics of viral and bacterial communities, with low bulk and cell-specific bacterial metabolism in winter, and a dramatic increase in injured bacteria under the ice cover in all lakes. This period was accompanied by contrasting patterns in the proportion of lysogenic cells. In the eutrophic lake, times of low bacterial metabolic rates and high proportion of damaged cells corresponded to highest levels of lysogeny, supporting the notion that hosts are a 'refuge' for viruses. In the two unproductive lakes, peaks of injured cells corresponded to a minimum of lysogeny, suggesting an 'abandon the sinking ship' response, where the prophage replicates before the loss of genome. We suggest that these diverging responses to the host physiological state are not contradictory, but rather that there may be thresholds of cell stress and metabolic activity leading to one or the other response. PMID- 20002138 TI - Phenotypic and genomic diversity of Lactobacillus plantarum strains isolated from various environmental niches. AB - Lactobacillus plantarum is a ubiquitous microorganism that is able to colonize several ecological niches, including vegetables, meat, dairy substrates and the gastro-intestinal tract. An extensive phenotypic and genomic diversity analysis was conducted to elucidate the molecular basis of the high flexibility and versatility of this species. First, 185 isolates from diverse environments were phenotypically characterized by evaluating their fermentation and growth characteristics. Strains clustered largely together within their particular food niche, but human fecal isolates were scattered throughout the food clusters, suggesting that they originate from the food eaten by the individuals. Based on distinct phenotypic profiles, 24 strains were selected and, together with a further 18 strains from an earlier low-resolution study, their genomic diversity was evaluated by comparative genome hybridization against the reference genome of L. plantarum WCFS1. Over 2000 genes were identified that constitute the core genome of the L. plantarum species, including 121 unique L. plantarum-marker genes that have not been found in other lactic acid bacteria. Over 50 genes unique for the reference strain WCFS1 were identified that were absent in the other L. plantarum strains. Strains of the L. plantarum subspecies argentoratensis were found to lack a common set of 24 genes, organized in seven gene clusters/operons, supporting their classification as a separate subspecies. The results provide a detailed view on phenotypic and genomic diversity of L. plantarum and lead to a better comprehension of niche adaptation and functionality of the organism. PMID- 20002141 TI - Crenarchaeota affiliated with group 1.1b are prevalent in coastal mineral soils of the Ross Sea region of Antarctica. AB - The objective of this study was to examine the presence and diversity of Archaea within mineral and ornithogenic soils from 12 locations across the Ross Sea region. Archaea were not abundant but DNA sufficient for producing 16S rRNA gene clone libraries was extracted from 18 of 51 soil samples, from four locations. A total of 1452 clones were analysed by restriction fragment length polymorphism and assigned to 43 operational taxonomic units from which representatives were sequenced. Archaea were primarily restricted to coastal mineral soils which showed a predominance of Crenarchaeota belonging to group 1.1b (> 99% of clones). These clones were assigned to six clusters (A through F), based on shared identity to sequences in the GenBank database. Ordination indicated that soil chemistry and water content determined archaeal community structure. This is the first comprehensive study of the archaeal community in Antarctic soils and as such provides a reference point for further investigation of microbial function in this environment. PMID- 20002140 TI - An alpha-amylase is a novel receptor for Bacillus thuringiensis ssp. israelensis Cry4Ba and Cry11Aa toxins in the malaria vector mosquito Anopheles albimanus (Diptera: Culicidae). AB - Bacillus thuringiensis ssp. israelensis (Bti) produces four Cry toxins (Cry4Aa, Cry4Ba, Cry10Aa and Cry11Aa), and two Cyt proteins (Cyt1Aa and Cyt2Ba), toxic to mosquito-larvae of the genus Aedes, Anopheles and Culex, important human disease vectors that transmit dengue virus, malaria and filarial parasites respectively. Previous work showed that Bti is highly toxic to Anopheles albimanus, the main vector for transmission of malaria in Mexico. In this work, we analysed the toxicity of isolated Cry proteins of Bti and identified an An. albimanus midgut protein as a putative Cry4Ba and Cry11Aa receptor molecule. Biossays showed that Cry4Ba and Cry11Aa of Bti are toxic to An. albimanus larvae. Ligand blot assays indicated that a 70 kDa glycosylphosphatidylinositol-anchored protein present in midgut brush border membrane vesicles of An. albimanus interacts with Cry4Ba and Cry11Aa toxins. This protein was identified as an alpha-amylase by mass spectrometry and enzymatic activity assays. The cDNA that codes for the alpha amylase was cloned by means of 5'- and 3'-RACE experiments. Recombinant alpha amylase expressed in Escherichia coli specifically binds Cry4Ba and Cry11Aa toxins. PMID- 20002142 TI - Examining the prevalence of epilepsy and delivery of epilepsy care in Ireland. AB - PURPOSE: To examine the prevalence of epilepsy and delivery of epilepsy care in Ireland. METHODS: A multiple case ascertainment methodology was employed examining five nationwide data sources. Data were obtained via (1) a nationwide population-based health survey, (2) secondary data analysis of all reimbursements for prescriptions of antiepilepsy medication nationwide from 2002 to 2005, (3) a nationwide survey of primary care community-based physicians, (4) a nationwide survey of specialist care hospital-based neurologists, and (5) secondary data analysis of inpatient discharges from acute hospitals nationwide for persons with a diagnosis of epilepsy between 2000 and 2005. RESULTS: The findings indicate that nationwide approximately (1) 10 per 1,000 persons (n = 31,000) 18 years and older have self-reported lifetime prevalence of epilepsy, (2) 8.3-9 per 1,000 (approximately 33,000-36,000) persons 5 years and older are being treated for epilepsy using antiepilepsy medication, (3) the typical general practitioner provides care to an average of 13 patients with active epilepsy and epilepsy in remission with treatment, (4) specialist care is provided on a weekly basis to approximately 442 persons with active epilepsy and epilepsy in remission with treatment, and (5) approximately 67 discharges for persons with a primary or secondary diagnosis of epilepsy are reported from acute hospitals weekly. DISCUSSION: This study is distinguished as the first nationwide prevalence study of epilepsy in Europe and provides previously unavailable data on this population in Ireland. Moreover, this study illustrates the merits of secondary data analysis of existing sources when access to clinical sources for the purpose of identifying epidemiologic samples is limited. PMID- 20002143 TI - Involvement of the thalamocortical network in TLE with and without mesiotemporal sclerosis. AB - PURPOSE: The thalamus plays an important role in seizure propagation in temporal lobe epilepsy (TLE). This study investigated how structural abnormalities in the focus, ipsilateral thalamus and extrafocal cortical structures relate to each other in TLE with mesiotemporal sclerosis (TLE-MTS) and without hippocampal sclerosis (TLE-no). METHODS: T1 and high-resolution T2 images were acquired on a 4T magnet in 29 controls, 15 TLE-MTS cases, and 14 TLE-no. Thalamus volumes were obtained by warping a labeled atlas onto each subject's brain. Deformation-based morphometry was used to identify regions of thalamic volume loss and FreeSurfer for cortical thickness measurements. CA1 volumes were obtained from high resolution T2 images. Multiple regression analysis and correlation analyses for voxel- and vertex-based analyses were performed in SPM2 and FreeSurfer. RESULTS: TLE-MTS had bilateral volume loss in the anterior thalamus, which was correlated with CA1 volume and cortical thinning in the mesiotemporal lobe. TLE-no had less severe volume loss in the dorsal lateral nucleus, which was correlated with thinning in the mesiotemporal region but not with extratemporal thinning. DISCUSSION: The findings suggest that seizure propagation from the presumed epileptogenic focus or regions close to it into the thalamus occurs in TLE-MTS and TLE-no and results in circumscribed neuronal loss in the thalamus. However, seizure spread beyond the thalamus seems not to be responsible for the extensive extratemporal cortical abnormalities in TLE. PMID- 20002144 TI - Inhibition of long-term potentiation by valproic acid through modulation of cyclic AMP. AB - PURPOSE: Valproic acid (VPA) is widely used clinically in epilepsy, bipolar disorder, and migraine. In experimental models, it has also been shown to have neuroprotective and antiepileptogenic effects. Its mechanisms of action in these diverse conditions are, however, unclear, but there is some evidence indicating an effect of VPA upon protein kinase A (PKA) activity. We, therefore, asked whether VPA modulates cyclic adenosine monophosphate (cAMP)/PKA-dependent synaptic plasticity and whether this mode of action could explain its anticonvulsant effect. METHODS: We first tested the effects of VPA on PKA dependent synaptic plasticity at mossy fiber to CA3 synapses in rat hippocampus slices following very high-frequency stimulation or application of the adenylyl cyclase activator forskolin. Using biochemical assays, we then tested whether VPA had a direct effect on PKA activity or an indirect effect through modulating cAMP production. Lastly, VPA and inhibitors of adenylyl cyclase (SQ22536) and PKA (H89) were tested in in vitro models of epileptiform activity induced in hippocampal-entorhinal cortex slices using either pentylenetetrazol (2 mM) or low magnesium. RESULTS: VPA (1 mm) inhibited PKA-dependent long-term potentiation of mossy fiber to CA3 pyramidal cell transmission. However, VPA did not directly modulate PKA activity but rather inhibited the accumulation of cAMP. In acute in vitro seizure models, the anticonvulsant activity of VPA is not mediated through modulation of adenylyl cyclase or PKA. CONCLUSIONS: These results indicate that VPA through an action on cAMP accumulation can inhibit synaptic plasticity, but this cannot fully explain its anticonvulsant effect. PMID- 20002145 TI - Prevalence of epileptiform discharges in healthy children--new data from a prospective study using digital EEG. AB - PURPOSE: Data on epileptiform electroencephalography (EEG) discharges in healthy children are limited, with published studies dating back more than 20 years. Moreover, analyses have been performed exclusively using paper-recorded EEG, and reported prevalences differ significantly. With recent reports using these data as reference suggesting an increased prevalence of epileptiform EEG discharges in children with behavioral disturbances, acquisition of exact prevalence data has become even more critical. The aim of our study was to analyze the frequency of epileptiform EEG discharges in healthy children using digitally recorded EEG (DEEG) and to compare these data to those of previously published studies. METHODS: Prospective analysis of DEEG was performed in 382 healthy children (226 male, 156 female) ages 6-13 years admitted to our hospital for minor head trauma. Recording was carried out for a minimum of 20 min including hyperventilation and photic stimulation. Analysis was carried out by two board-certified clinical neurophysiologists. RESULTS: Epileptiform EEG discharges were detected in 25 of 382 children (11 of 226 male, 14 of 156 female) corresponding to an overall prevalence of 6.5%. Of these 25 children, 4 had either generalized or bifrontal spikes, 12 showed constant localized focal discharges, and 9 showed multifocal discharges. Compared to previous studies using non-DEEG recording, the prevalence of epileptiform EEG discharges in our population was significantly higher. No significant difference was found when comparing our data to prevalences recently reported in children with behavioral disturbances using DEEG. CONCLUSIONS: Our study further highlights the urgent need to reevaluate the prevalence of epileptiform EEG discharges in healthy children using DEEG recordings in a large cohort. PMID- 20002146 TI - De novo epileptic confusion in the elderly: a 1-year prospective study. AB - PURPOSE: Nonconvulsive status epilepticus (NCSE) is clinically difficult to diagnose, especially in old patients without epilepsy, and requires electroencephalography (EEG) for diagnosis. Its incidence among elderly patients with confusion of unknown origin (CUO) remains undetermined. METHODS: A 1-year prospective study was conducted in patients aged 60 years or older, for whom EEG was requested because of confusion considered to be of unknown origin after initial clinical, biologic, and imaging investigations. Diagnosis criteria included a validated clinical assessment scale to confirm confusion. RESULTS: Of 44 patients with confirmed CUO, 7 presented with de novo NCSE. NCSE population had a mean age of 76 years (range, 60-97 years). No statistically significant differences were found between NCSE patients and others for age, drugs, presence of myoclonia, eyelid myoclonia, tachycardia, or agitation. In contrast, an acute onset (<24 h), gender (100% female among NCSE patients), and lack of clinical response to simple commands were significantly associated with NCSE. No differences between the two groups were evidenced for mortality and morbidity (length of hospitalization, social outcome, and so on). DISCUSSION: Almost 16% of patients aged 60 or older with confusion of unknown origin had NCSE, according to this first prospective study. Female patients with rapid onset (<24 h) of symptoms and lack of response to simple commands were at a higher risk of presenting with NCSE. PMID- 20002147 TI - Prognosticating acute symptomatic seizures using two different seizure outcomes. AB - PURPOSE: This study examined the profiles and prognosis of first acute symptomatic seizure (ASS). Because seizure recurrences may occur in the setting of a persisting or reemerging acute symptomatic cause or in the setting of an unprovoked seizure, we documented the prognosis of ASS in terms of acute symptomatic seizure (AS) or unprovoked seizure (US) recurrence. METHODS: We conducted a prospective study of patients with suspected seizures between April 2004 and December 2005. Patients were classified according to medical history taking, routine clinical evaluation, and expert adjudication, and they were followed for a minimum of 2 years or until death. The Kaplan-Meier method and univariate/multivariate statistical analysis were used to determine prognosis. RESULTS: One hundred five patients with first-ever ASS were identified. For many, first ASS was associated with status epilepticus (29.5%), multiple-onset (>1 seizure within 24 h on day of presentation) (35.2%), and multiple etiologies (22.9%), with a mortality of 30% at 2 years (Kaplan-Meier method). Using AS as outcome, the risk of recurrence following an ASS was 32% at 2 years [mean time to recurrence 20.5 days with epileptiform electroencephalography (EEG) being an independent predictor; p = 0.005, odds ratio (OR) 16, 95% confidence interval (CI) 4.09-62.7]. Using US as outcome, the risk of recurrence following an ASS was 12% at 2 years. DISCUSSION: Although ASS did not associate with a high rate of US recurrence, we demonstrated that ASS was often followed by another AS. This may have implication for short- to medium-term antiepileptic agent therapy, especially when the acute symptomatic cause takes a long time to treat, is prone to reemergence, or is irreversible. PMID- 20002148 TI - Theiler's virus infection chronically alters seizure susceptibility. AB - PURPOSE: Central nervous system infections greatly increase the risk for the development of seizures and epilepsy (recurrent unprovoked seizures). We have previously shown that Theiler's murine encephalomyelitis virus (Theiler's virus or TMEV) infection causes acute symptomatic seizures in C57BL/6 (B6) mice. The objective of the present study was threefold: (1) to assess pathologic changes associated with acute TMEV infection and infection-induced seizures, (2) to determine whether Theiler's virus infection and associated acute seizures lead to chronically altered seizure susceptibility, and (3) to determine whether genetic background influences seizure susceptibility following Theiler's virus infection. METHODS: Immunohistochemical techniques were used to assess Theiler's virus antigen localization in the brain and associated neuronal cell death. A battery of electroconvulsive threshold (ECT) tests and corneal kindling studies were conducted to assess whether there were chronic alterations in seizure susceptibility and kindling development. Studies were conducted in both B6 and SJL/J mice to assess strain-dependent effects. RESULTS: Histopathologic analyses indicate that TMEV has specific tropism for limbic structures and causes widespread cell death in these regions. Results from ECT studies demonstrate that B6 mice that displayed acute symptomatic seizures have significantly reduced seizure thresholds and kindle faster than either control mice or infected mice without acute seizures. Furthermore, these effects were mouse-strain dependent, since SJL/J mice displayed a different seizure threshold spectrum. DISCUSSION: These findings indicate that Theiler's virus infection leads to chronically altered seizure susceptibility in mice. It is important to note that Theiler's virus infection of B6 mice represents a novel model to study postinfection hyperexcitability. PMID- 20002149 TI - Health care resource utilization in patients with active epilepsy. AB - PURPOSE: To evaluate health care resource utilization (HRU) in active epilepsy. METHODS: Thomson-Reuters insurance databases included 14 million persons in 2005 2007. We extracted information for individuals with insurance claims suggestive of epilepsy. Using iterative expert classification, we sorted patients by type of epilepsy. For each type we calculated prevalence and HRU. A distance analysis identified closely similar types, and a principal components analysis revealed dimensions of variation in HRU. RESULTS: The prevalence of active epilepsy was 3.4 per 1,000. Most common diagnoses among 46,847 patients were generalized convulsive epilepsy (33.3%) and complex partial seizures (24.8%). Patients averaged 10 physician visits per year, 24 diagnostic tests/procedures per year, >30 drug dispensings per year, and <1 emergency room (ER) visit per year, the minority of each of these being related to epilepsy. Female patients generally had more HRU, and HRU increased with age. Patients were hospitalized most frequently for disorders other than epilepsy. HRU was similar for most epilepsy types, excepting grand mal status, epilepsia partialis continua, and infantile spasms. The first principal components of HRU variation was nonepilepsy HRU, followed by components of epilepsy-related medications, other epilepsy/emergency care, and epilepsy visits/diagnostic procedures. DISCUSSION: The prevalence of active epilepsy in the United States is substantially less than the prevalence of any history of recurrent seizure. Nonepilepsy-related HRU dominated HRU in epilepsy patients and was the principal source of variation. There is a core set of epilepsy diagnoses, the HRU patterns of which are indistinguishable, whereas patients with grand mal status, epilepsia partialis continua, and infantile spasms all have distinct patterns. To provide more specific insights into the economic impact of the condition, studies of HRU in epilepsy should make a distinction about epilepsy-related and unrelated care. PMID- 20002150 TI - Direct and indirect costs associated with epileptic partial onset seizures among the privately insured in the United States. AB - PURPOSE: Compare annual direct and indirect costs between privately insured U.S. patients with epileptic partial onset seizures (POS) and matched controls. METHODS: One thousand eight hundred fifty-nine patients (including a subset of 758 employees) with >or=1 (POS) diagnosis (ICD-9-CM: 345.4.x-345.7.x), 1999-2004, ages 16-64 years, were identified from a privately insured claims database. Control group was an age- and gender-matched cohort of randomly chosen beneficiaries without epilepsy (ICD-9-CM: 345.x). All were required to have continuous health coverage during 2004 (baseline) and 2005 (study period). Chi square tests were used to compare baseline comorbidities. Univariate and multivariate analyses were used for comparisons of annual direct (medical and pharmaceutical) and indirect costs during the study period. RESULTS: Patients with POS were on average 42 years of age, and 57% were women. Patients with POS had significantly higher rates of mental health disorders, migraine, and other neurologic disorders, and higher Charlson comorbidity index (CCI) compared with controls. On average, direct annual costs were significantly higher for POS patients ($11,276) compared with controls ($4,087), p < 0.001; difference of $7,190. Epilepsy-related costs (i.e., costs for antiepileptic drugs, claims with epilepsy or convulsions diagnoses) accounted for $3,290 (29% of direct costs). Employees with POS had substantial and significantly higher indirect (disability- and medically related absenteeism) costs compared with controls ($3,431 vs. $1,511, p < 0.001). Multivariate analyses supported the matched-control univariate findings. CONCLUSION: Patients with POS had significantly higher costs compared with matched controls. Epilepsy-related costs underestimate the excess costs of patients with partial onset seizures. PMID- 20002151 TI - Cortical stimulation for language mapping in focal epilepsy: correlations with tractography of the arcuate fasciculus. AB - PURPOSE: Diffusion tensor imaging (DTI) provides information about magnitude (diffusivity) and directionality (fractional anisotropy, FA) of water diffusion and allows visualization of major white matter tracts. The arcuate fasciculus (AF) connects anterior (Broca's) and posterior (Wernicke's) language areas. We hypothesized that essential language areas identified by direct cortical stimulation would colocalize with areas revealing subcortical connectivity via the AF. METHODS: Fourteen patients with pharmacoresistant left hemispheric epilepsy and left hemisphere language dominance underwent invasive evaluations for localization of epileptogenicity and functional mapping. DTI and T1-weighted volumetric magnetic resonance imaging (MRI) scans were coregistered, and subdural grid electrodes identified on postimplantation computed tomography (CT) scans were also coregistered to the MRI scans. The AF was reconstructed from a region lateral to the corona radiata on the FA map. Colocalization, defined as <1 cm between the AF and the electrode positions delineating language cortex, was visually assessed with excellent reliability (Cronbach's alpha = 0.98). RESULTS: A total of 71 subdural grid contacts were overlying language cortex. Nineteen contacts in eight patients were over Broca's area, 16 of which (84.2%) colocalized with the AF. Fifty-two contacts in 10 patients were over Wernicke's area, with colocalization in 29 patients (55.8%). Colocalization was significantly greater in anterior regions than in posterior regions [chi(2)(1) = 4.850, p < 0.05]. DISCUSSION: The AF, as visualized with DTI, colocalized well with anterior language areas, but less so with posterior language areas, inferring that the latter are more spatially dispersed. PMID- 20002152 TI - Spatiotemporal patterns of electrocorticographic very fast oscillations (> 80 Hz) consistent with a network model based on electrical coupling between principal neurons. AB - PURPOSE: We sought to characterize spatial and temporal patterns of electrocorticography (ECoG) very fast oscillations (> ~80 Hz, VFOs) prior to seizures in human frontotemporal neocortex, and to develop a testable network model of these patterns. METHODS: ECoG data were recorded with subdural grids from two preoperative patients with seizures of frontal lobe onset in an epilepsy monitoring unit. VFOs were recorded from rat neocortical slices. A "cellular automaton" model of network oscillations was developed, extending ideas of Traub et al. (Neuroscience, 92, 1999, 407) and Lewis & Rinzel (Network: Comput Neural Syst, 11, 2000, 299); this model is based on postulated electrical coupling between pyramidal cell axons. RESULTS: Layer 5 of rat neocortex, in vitro, can generate VFOs when chemical synapses are blocked. Human epileptic neocortex, in situ, produces preseizure VFOs characterized by the sudden appearance of "blobs" of activity that evolve into spreading wavefronts. When wavefronts meet, they coalesce and propagate perpendicularly but never pass through each other. This type of pattern has been described by Lewis & Rinzel in cellular automaton models with spatially localized connectivity, and is demonstrated here with 120,000- to 5,760,000-cell models. We provide a formula for estimating VFO period from structural parameters and estimate the spatial scale of the connectivity. DISCUSSION: These data provide further evidence, albeit indirect, that preseizure VFOs are generated by networks of pyramidal neurons coupled by gap junctions, each predominantly confined to pairs of neurons having somata separated by < ~1-2 mm. Plausible antiepileptic targets are tissue mechanisms, such as pH regulation, that influence gap-junction conductance. PMID- 20002153 TI - Hippocampal malrotation in pediatric patients with epilepsy associated with complex prefrontal dysfunction. AB - PURPOSE: The cognitive consequences of hippocampal malrotation (HIMAL) were investigated in a matched control study of children with epilepsy. METHODS: Seven children with HIMAL were compared on a range of memory and attention tasks with 21 control children with epilepsy without temporal role pathology and 7 children with epilepsy and magnetic resonance imaging (MRI)-documented hippocampal sclerosis. In addition, in a statistical morphometric analysis, MRI studies from four children with HIMAL were compared to similar images of 20 age-matched typically developing control children. RESULTS: Although the task battery was sensitive to the memory deficit of the children with hippocampal sclerosis, it did not reveal memory impairment in the patients with HIMAL. In contrast, the patients with HIMAL were impaired on the attentionally more demanding dual tasks, compared to both the control and the hippocampal sclerosis group. The structural MRI analysis revealed morphometric abnormalities in the tail of the affected hippocampus, the adjacent neocortex, and the ipsilateral medial thalamus. The basal forebrain was bilaterally affected. Abnormalities in remote cortex were found in the ipsilateral temporal lobe, the contralateral anterior cingulate gyrus, and bilateral in the dorsolateral and lateral-orbitofrontal prefrontal cortex. DISCUSSION: Because the prefrontal cortical regions have been shown to be active during dual-task performance, the MRI results converge with the neuropsychological findings of impairment on these tasks. We conclude that HIMAL had no direct memory repercussions, but was secondary to subtle but widespread neurologic abnormalities that also affected morphology and functioning of the prefrontal cortex. PMID- 20002154 TI - Chromosome 1 abnormalities in myeloid malignancies: a literature survey and karyotype-phenotype associations. AB - Chromosome 1 is the largest human chromosome and contains over 1600 known genes and 1000 novel coding sequences or transcripts. It is, therefore, not surprising that recurrent chromosome 1 abnormalities are regularly encountered in both neoplastic and non-neoplastic medical conditions. The current review is focused on myeloid malignancies where we summarize the relevant published literature and discuss specific karyotype-phenotype associations. We show that chromosome 1 abnormalities are most frequent in BCR-ABL-negative classic myeloproliferative neoplasms (MPN): polycythemia vera (PV), essential thrombocythemia (ET), and primary myelofibrosis (PMF). Specific abnormalities include duplications (e.g. 1q12-->1q32 in PV, 1q21-32-->1q32-44 in post-PV MF or PMF), deletions (e.g. 1p13 36-->pter in PV or PMF, 1q21 in PMF) and unbalanced translocations involving chromosome 6, such as der(6)t(1;6)(q21-25;p21.3-23), and other partner chromosomes involving 1q10/1p11 and 1q21-25 breakpoints. Although occasionally seen in chronic phase MPN, unbalanced 1;7 translocations, e.g. der(1;7)(q10;p10), are usually seen in acute myeloid leukemia (AML), myelodysplastic syndromes (MDS), and post-MPN AML/MDS. These observations suggest that certain chromosome 1 regions, especially 1q21-1q32 and 1p11-13, might harbor oncogenes or tumor suppressor genes that are pathogenetically relevant to both chronic and advanced phases of MPN. PMID- 20002155 TI - Diagnostic challenges and recent advances in the early management of invasive fungal infections. AB - During the past 20 yr, the population of immunocompromized patients at risk of developing invasive fungal infections (IFIs) has increased, and there has been a shift in fungal epidemiology, with more infections caused by non-Aspergillus molds and yeasts, which are often resistant to one or more antifungal drugs. Traditional diagnostic methods, such as culture and the histopathology of infected tissue, often fail to detect IFIs until the later stages. Furthermore, invasive diagnostic methods to obtain tissue may be contraindicated in severely ill patients; even when tissue is available, the morphology of several filamentous fungi is identical, or the cultures may fail to grow the pathogen. Recently developed non-invasive diagnostic techniques, such as tests for serum markers and polymerase chain reaction assays, may allow for earlier and more accurate diagnoses - crucial in the effort to reduce morbidity and the risk of mortality. This article reviews current approaches to diagnosis and treatment, focusing on how an early and accurate diagnosis can guide treatment and improve outcomes. Strategies for improving the management of IFIs also are discussed. PMID- 20002156 TI - GVHD prophylaxis using low-dose cyclosporine improves survival in leukaemic recipients of HLA-identical sibling transplants. AB - Graft-versus-host disease (GVHD) prophylaxis of short duration (6 months) with low-dose cyclosporine A (CsA) starting at 1 mg/kg per day i.v. and four doses of methotrexate (MTX) were given to 171 consecutive leukaemic recipients of HLA identical sibling transplants. In contrast, apart from MTX, retrospective controls received high-dose CsA, starting at 5-7.5 mg/kg per day i.v. and discontinued 1 yr post-transplant. In the low-dose CsA group, the probability of acute GVHD grades I-II (70% vs. 53%, P < 0.01), and chronic GVHD were increased (58% vs. 25%, P < 0.01), whereas the incidences of acute GVHD grades III-IV (9% vs. 5%, P = 0.62), and non-relapse mortality (20% vs. 22%, P = 0.58) were similar. Moreover, the probability of relapse was decreased (31% vs. 54%, P < 0.01), and both relapse-free (56% vs. 38%, P = 0.04) and overall survival (61% vs. 40%, P = 0.04) were markedly improved using the low-dose CsA regimen. In multivariate analyses, low-dose CsA was strongly associated with chronic GVHD [relative hazard (RH) 2.56, P < 0.01], which decreased the risk of relapse (RH 0.46, P < 0.01) and improved the probability of survival (RH 1.84, P < 0.01). In conclusion, a low-dose CsA regimen in leukaemic recipients of HLA-identical sibling transplants increases the rate of chronic GVHD, which seems to attenuate the risk of relapse, thereby improving patient survival owing to enhanced graft versus-leukaemia effect. PMID- 20002157 TI - Epigenetic inactivation of tumour suppressor gene KLF11 in myelodysplastic syndromes*. AB - The identification of aberrantly hypermethylated genes may lead to the development of new diagnostic markers and the identification of novel targets of epigenetic therapy in myelodysplastic syndromes (MDS). We therefore investigated the methylation status of transcription factor genes KLF5, KLF11, and MAFB, shown to be aberrantly methylated in myelogeneous leukaemia cells, in a series of 115 MDS patient as well as in 25 control subjects. Using quantitative high-resolution pyrosequencing methodology, KLF11, MAFB, and KLF5 were shown for the first time to be hypermethylated in 17 (15%), 8 (7%), and 2 (1.7%) cases, respectively, but not in any of the patients with an isolated 5q-deletion. Patient samples harbouring KLF11 methylation displayed reduced KLF11 mRNA expression and KLF11 hypermethylation correlated with a high International Prognostic Scoring System score (P < 0.05). In conclusion, epigenetic inactivation and subsequent transcriptional repression of the KLF11 gene is quite frequent in MDS. Patients with an isolated 5q-deletion seem to harbour a distinct epigenetic profile. PMID- 20002159 TI - BCR-ABL-independent and RAS / MAPK pathway-dependent form of imatinib resistance in Ph-positive acute lymphoblastic leukemia cell line with activation of EphB4. AB - OBJECTIVE: We investigated the mechanism responsible for imatinib (IM) resistance in Philadelphia chromosome-positive acute lymphoblastic leukemia (Ph(+) ALL) cell lines. METHODS: We established cell lines from a patient with Ph(+) ALL at the time of first diagnosis and relapsed phase and designated as NPhA1 and NPhA2, respectively. We also derived IM-resistant cells, NPhA2/STIR, from NPhA2 under gradually increasing IM concentrations. RESULTS: NPhA1 was sensitive to IM (IC(50) 0.05 microm) and NPhA2 showed mild IM resistance (IC(50) 0.3 microm). NPhA2/STIR could be maintained in the presence of 10 microm IM. Phosphorylation of MEK and ERK was slightly elevated in NPhA2 and significantly elevated in NPhA2/STIR compared to NPhA1 cells. After treatment with IM, phosphorylation of MEK and ERK was not suppressed but rather increased in NPhA2 and NPhA2/STIR. Active RAS was also increased markedly in NPhA2/STIR after IM treatment. The expression of BCL-2 was increased in NPhA2 compared to NPhA1, but no further increase in NPhA2/STIR. Proliferation of NPhA2/STIR was significantly inhibited by a combination of MEK inhibitor and IM. Analysis of tyrosine phosphorylation status with a protein tyrosine kinase array showed increased phosphorylation of EphB4 in NPhA2/STIR after IM treatment. Although transcription of EphB4 was suppressed in NPhA1 and NPhA2 after IM treatment, it was not suppressed and its ligand, ephrinB2, was increased in NPhA2/STIR. Suppression of EphB4 transcripts by introducing short hairpin RNA into NPhA2/STIR partially restored their sensitivity to IM. CONCLUSIONS: These results suggest a new mechanism of IM resistance mediated by the activation of RAS/MAPK pathway and EphB4. PMID- 20002161 TI - Intergenomic epistasis and coevolutionary constraint in plants and rhizobia. AB - Studying how the fitness benefits of mutualism differ among a wide range of partner genotypes, and at multiple spatial scales, can shed light on the processes that maintain mutualism and structure coevolutionary interactions. Using legumes and rhizobia from three natural populations, I studied the symbiotic fitness benefits for both partners in 108 plant maternal family by rhizobium strain combinations. Genotype-by-genotype (G x G) interactions among local genotypes and among partner populations determined, in part, the benefits of mutualism for both partners; for example, the fitness effects of particular rhizobium strains ranged from uncooperative to mutualistic depending on the plant family. Correlations between plant and rhizobium fitness benefits suggest a trade off, and therefore a potential conflict, between the interests of the two partners. These results suggest that legume-rhizobium mutualisms are dynamic at multiple spatial scales, and that strictly additive models of mutualism benefits may ignore dynamics potentially important to both the maintenance of genetic variation and the generation of geographic patterns in coevolutionary interactions. PMID- 20002162 TI - Maternal effects, but no good or compatible genes for sperm competitiveness in Australian crickets. AB - Explanations for the evolution of polyandry often center on the idea that females garner genetic benefits for their offspring by mating multiply. Furthermore, postcopulatory processes are thought to be fundamental to enabling polyandrous females to screen for genetic quality. Much attention has focused on the potential for polyandrous females to accrue such benefits via a sexy- or good sperm mechanism, whereby additive variation exists among males in sperm competitiveness. Likewise, attention has focused on an alternative model, in which offspring quality (in this context, the sperm competitiveness of sons) hinges on an interaction between parental haplotypes (genetic compatibility). Sperm competitiveness that is contingent on parental compatibility will exhibit nonadditive genetic variation. We tested these models in the Australian cricket, Teleogryllus oceanicus, using a design that allowed us to partition additive, nonadditive genetic, and parental variance for sperm competitiveness. We found an absence of additive and nonadditive genetic variance in this species, challenging the direct relevance of either model to the evolution of sperm competitiveness in particular, and polyandry in general. Instead, we found maternal effects that were possibly sex-linked or cytoplasmically linked. We also found effects of focal male age on sperm competitiveness, with small increments in age conferring more competitive sperm. PMID- 20002163 TI - Is the age of the Earth one of our "sorest troubles?" students' perceptions about deep time affect their acceptance of evolutionary theory. PMID- 20002164 TI - Z linkage of female promiscuity genes in the moth Utetheisa ornatrix: support for the sexy-sperm hypothesis? AB - Female preference genes for large males in the highly promiscuous moth Utetheisa ornatrix (Lepidoptera: Arctiidae) have previously been shown to be mostly Z linked, in accordance with the hypothesis that ZZ-ZW sex chromosome systems should facilitate Fisherian sexual selection. We determined the heritability of both female and male promiscuity in the highly promiscuous moth U. ornatrix (Lepidoptera: Arctiidae) through parent-offspring and grandparent-offspring regression analyses. Our data show that male promiscuity is not sex-limited and either autosomal or sex-linked whereas female promiscuity is primarily determined by sex-limited, Z-linked genes. These data are consistent with the "sexy-sperm hypothesis," which posits that multiple-mating and sperm competitiveness coevolve through a Fisherian-like process in which female promiscuity is a kind of mate choice in which sperm-competitiveness is the trait favored in males. Such a Fisherian process should also be more potent when female preferences are Z-linked and sex-limited than when autosomal or not limited. PMID- 20002165 TI - The measurement theory of fitness. AB - In this article, an approach to measure fitness is proposed that considers fitness as a measure of competitive ability among phenotypes or genotypes. This approach is based on pairwise competition tests and is related to measures of "utility" in mathematical economics. Extending the results from utility theory it is possible to recover the classical Wrightian fitness measure without reference to models of population growth. A condition, quasi-BTL, similar to the Bradley Terry-Luce condition of classical utility theory is shown to be necessary for the existence of frequency and context-independent fitness measures. Testing for violations of this quasi-BTL condition can be used to the detect genotype-by genotype interactions and frequency-dependent fitness. A method for the detection of genotype by environment interactions is proposed that avoids potential scaling artifacts. Furthermore the measurement theoretical approach allows one to derive Wright's selection equation. This shows that classical selection equations are entirely general and exact. It is concluded that measurement theory is able to give definite answers to a number theoretical and practical questions. For instance, this theory identifies the correct scale for measuring gene interaction with respect to fitness and shows that different scales may lead to wrong conclusions. PMID- 20002166 TI - The origin of polymorphic crypsis in a heterogeneous environment. AB - Polymorphic crypsis has been observed in several taxa, but has, until now, lacked a firm theoretical understanding. How does a single morph, well camouflaged in one type of habitat, evolve crypsis in another, not isolated, habitat? We here analyze a model of one prey species living in two different habitats connected by passive dispersal. We find that the rate of dispersal, the trade-off between crypticity in the habitats, and the amount of predation determines whether the prey species can become cryptic in two different habitats through evolutionary branching. Intermediate values of all parameters seem to promote evolutionary branching leading to polymorphism, and a more extreme value of one parameter can be balanced by another. Other parameter combinations lead to either a single habitat specialist or an intermediate generalist type, partly cryptic in both habitats. When the predator follows a type III functional response, the parameter space for when the prey will undergo evolutionary branching is remarkably larger than the corresponding parameter space for a type II functional response. Evolutionary branching can occur both at the intermediate generalist strategy, or close to a specialist strategy. PMID- 20002167 TI - Allopatric divergence and speciation in coral reef fish: the three-spot dascyllus, Dascyllus trimaculatus, species complex. AB - Long pelagic larval phases and the absence of physical barriers impede rapid speciation and contrast the high diversity observed in marine ecosystems such as coral reefs. In this study, we used the three-spot dascyllus (Dascyllus trimaculatus) species complex to evaluate speciation modes at the spatial scale of the Indo-Pacific. The complex includes four recognized species and four main color morphs that differ in distribution. Previous studies of the group using mitochondrial DNA revealed a noncongruence between color morphs and genetic groupings; with two of the color morphs grouped together and one color morph separated into three clades. Using extensive geographic sampling of 563 individuals and a combination of mitochondrial DNA sequences and 13 nuclear microsatellites, we defined population/species boundaries and inferred different speciation modes. The complex is composed of seven genetically distinct entities, some of which are distinct morphologically. Despite extensive dispersal abilities and an apparent lack of barriers, observed genetic partitions are consistent with allopatric speciation. However, ecological pressure, assortative mating, and sexual selection, were likely important during periods of geographical isolation. This study therefore suggests that primarily historical factors later followed by ecological factors caused divergence and speciation in this group of coral reef fish. PMID- 20002168 TI - Sex allocation based on relative and absolute condition. AB - Traditional models predict that organisms should allocate to sex based on their condition relative to the condition of their competitors, tracking shifts in mean condition in fluctuating environments, and maintaining an equilibrium sex ratio. In contrast, when individuals are constrained to define their condition absolutely, environmental fluctuations induce fluctuating sex ratios and the evolutionary loss of condition-dependent sex allocation in short-lived organisms. Here, we present a simulation model of temperature-dependent sex determination (TSD) in fluctuating environments that specifically examines the importance of relativity in defining individual condition. When relativity in condition is allowed to evolve, short-lived organisms evolve switchlike TSD reaction norms and define their condition relative to the annual temperature distribution, thus preventing biased cohort sex ratios in extreme years. Long-lived organisms also evolve switchlike reaction norms, but define condition less relatively and experience biased cohort sex ratios. The predictions are supported by data from painted turtles, where TSD reaction norms exhibit pivotal temperatures of sex determination that partially track mean annual temperature. Examining relativity in amniotic vertebrates provides a conceptual framework for multifactorial sex determination and suggests new ways of exploring adaptive hypotheses of sex allocation by focusing on the importance of frequency-dependent selection on sex. PMID- 20002169 TI - Platelet satellitism. PMID- 20002170 TI - Activating transcription factor 3 (ATF3) expression is increased in erythema multiforme and is regulated by IFN-gamma in human keratinocytes. AB - Activating transcription factor 3 (ATF3) expression is increased in erythema multiforme and is regulated by IFN-gamma in human keratinocytes. Experimental Dermatology 2010; 19: e310-e313. Abstract: Activating transcription factor 3 (ATF3) is a member of the ATF/cyclic AMP responsive element-binding protein (CREB) family of transcription factors and is involved in the regulation of immune responses, apoptosis, DNA repair and oncogenesis. The epidermal expression of ATF3 in the setting of cutaneous inflammation has not been well characterized. To examine the expression of ATF3 in the setting of inflammatory skin disease, ATF3 protein expression was analysed by immunohistochemistry (IHC). We found diffuse epidermal ATF3 protein expression in skin biopsies of erythema multiforme (EM). Given the role of interferon (IFN)-gamma in erythema multiforme, we sought to examine the impact of IFN-gamma on ATF3 expression in human keratinocytes. IFN gamma induced ATF3 mRNA and protein in primary human keratinocytes and HaCaT cells. Thus, epidermal ATF3 expression can be increased in the setting of inflammatory skin diseases and is regulated by IFN-gamma in human keratinocytes. PMID- 20002171 TI - Effects of metals on skin permeability barrier recovery. AB - We previously demonstrated that the electrical state of the skin surface influences epidermal permeability barrier homeostasis. At the interface between different materials, electrons are localized heterogeneously and induce electrical potential. In the present study, we evaluated the effects of metals on the barrier recovery. When we put pure gold plate on skin immediately after tape stripping, the barrier recovery rate was faster than the control. The acceleration of barrier recovery was blocked when the plate was earthed (grounded). When a plastic membrane was sandwiched between the plate and the skin, the recovery was delayed in comparison with the control. We then used a germanium diode to regulate the current flow between the plate and the earth. When the current was blocked, the barrier recovery was accelerated, but when the current was not blocked, the recovery was not accelerated. These results suggest that localization of electron might affect for the barrier recovery rate. The level of interfacial electric potential would be different due to the electrochemical property of metal. Thus, we next evaluated the effects of other metals. With samarium, zirconium, iridium and silver, the barrier recovery rate was faster than in the case of gold, while a platinum plate induced slower recovery than in the case of gold. There was a significant correlation between work function of each metal and barrier recovery rate. These results suggest that electron donation from outside accelerated the skin barrier recovery. PMID- 20002172 TI - Decreased expression of neurofibromin contributes to epithelial-mesenchymal transition in neurofibromatosis type 1. AB - Plexiform and/or dermal neurofibromas are nerve sheath tumors of the peripheral nervous system that are usually present in individuals with neurofibromatosis type 1 (NF1). Neurofibromas arise from Schwann cells with biallelic inactivation of NF1, the gene that encodes neurofibromin. This protein is responsible for regulation of the Ras-mediated pathway, which has been shown to play a crucial role in epithelial-to-mesenchymal transition (EMT). EMT is a biological process that occurs during embryogenesis and wound healing and is involved in pathological processes such as organ fibrosis and cancer metastasis. However, the relationship between neurofibromin and EMT has not been elucidated. We investigated whether the EMT-related signaling pathway was upregulated in NF1 associated neurofibromas and Schwann cells by assessing the expression levels of the EMT-related transcription factors Snail, Slug, Twist, ZEB1 and ZEB2. Immunohistochemical studies and quantitative reverse transcription polymerase chain reaction revealed an increase in the expression levels of EMT-related transcription factors in neurofibroma specimens and NF1-derived Schwann cells (sNF96.2). In addition, the silencing of NF1 by siRNA induced the expression of EMT-related transcription factors in normal human Schwann cells and in epithelial like breast cancer cells. Our findings suggest that the loss of neurofibromin activated the EMT-related signaling pathway and that the excessive mesenchymal reaction may play a key role in the development of NF1-associated neurofibromas. PMID- 20002173 TI - Alternate trafficking of cathepsin L in dermal fibroblasts induced by UVA radiation. AB - UVA radiation is increasingly used to treat fibrotic skin disorders. However, the mechanisms underlying the therapeutic effects of UVA for these disorders are only partially understood. Cathepsin L is a lysosomal cysteine protease, which has been shown to degrade various matrix proteins thus contributing to extracellular remodeling. Therefore, we investigated whether UVA irradiation regulates the expression and release of cathepsin L in human dermal fibroblasts. No alterations were found after single irradiation; however, a significantly increased extracellular release of cathepsin L was observed after repeated irradiation up to four times. The transcript levels of cathepsin L were elevated after repetitive irradiation, leading to increased amounts of total cathepsin L protein. Furthermore, higher amounts of extracellular cathepsin L were associated with a significant reduction of intracellular processed cathepsin L and an accumulation of unprocessed procathepsin L. The use of specific inhibitors elucidated mannose phosphate-independent sorting pathways of cathepsin L leading to enhanced secretion and reduced intracellular processing. This is the first study which demonstrates that alternate trafficking mechanisms mediate the extracellular release of a cysteine protease induced by repetitive UVA irradiation. PMID- 20002174 TI - Overexpression of connexin26 in the basal keratinocytes reduces sensitivity to tumor promoter TPA. AB - Connexin 26 is important in keratinocyte proliferation, differentiation and skin pathologies. Cx26 is barely expressed in normal adult epidermis, but its expression is induced during wound healing, psoriasis, and skin hyperplasia stimulated by tumor promoters. In hyperplastic proliferating epidermis, Cx26 is co-expressed with Cx43 typical for basal and suprabasal keratinocytes. As Cx26 and Cx43 can not form permeable gap junctions, their co-expression may alter the gap junctional communication between keratinocytes and induce proliferation. To test the effect of persistent co-expression of Cx26 and Cx43 in epidermis, we generated transgenic mice using keratin5 promoter to target Cx26 to basal Cx43 positive keratinocytes. We evaluated the effect of ectopic Cx26 on keratinocyte proliferation and differentiation in normal and 12-O-tetradecanoyl-phorbol-13 acetate (TPA)-treated skin. The ectopic Cx26 expression in epidermis did not significantly affect skin development, keratinocyte differentiation and proliferation in newborn and adult skin. Unexpectedly, the proliferative effect of tumor promoter TPA was strongly decreased in epidermis of K5.Cx26 transgenics. This correlated with significant down-regulation of TPA-induced activity of protein kinase C (PKC) in K5.Cx26 mice. PMID- 20002175 TI - Functional characterization of T cells differentiated in vitro from bone marrow derived CD34 cells of psoriatic patients with family history. AB - BACKGROUND: The strong but complex genetic background suggests that inherent and intrinsic rather than exogenous factors have a key role in immunopathogenesis of psoriasis. It is reasonable to speculate that the dysfunctional activity of psoriatic T cells may partly originate from the abnormal haematopoietic cells. OBJECTIVES: To test if T cells originated from haematopoietic progenitor cells in psoriasis patients display functional alternations similar to previously reported abnormalities of circulating T cells. METHODS: Bone marrow CD34(+) haematopoietic cells were isolated from psoriatic patients with family history and healthy subjects, and differentiated into T cells in vitro in the thymic stromal co culture system. These cells were further subjected to functional comparisons such as in vitro proliferation, secretion of cytokines such as IL-4, IL-8 and IFN gamma, and inducing the production of C-myc, Bcl-xL, and Ki67 proteins in human keratinocytes. RESULTS: While bone marrow-derived CD34(+) cells from both patients and healthy volunteers developed into mature T cells within weeks in the thymic environment in vitro, the differentiated T cells from psoriatic patients showed higher proliferation and stronger capacity to secret TH1 cytokines in response to streptococcal superantigen. The differentiated T cells from psoriatic patients, but not from normal controls, induced overexpression of C-myc and Ki67, but not Bcl-XL, in keratinocytes. CONCLUSIONS: T cells differentiated from CD34(+) cells of psoriatic patients, but not normal controls, are functionally similar to psoriatic circulating T cells, suggesting that the dysfunctional activity of T cells in psoriatic patients can be traced back to the early development of haematopoietic cells. PMID- 20002176 TI - Differences between tuberculosis cases infected with Mycobacterium africanum, West African type 2, relative to Euro-American Mycobacterium tuberculosis: an update. AB - Mycobacterium africanum (MAF) is a common cause of human pulmonary tuberculosis in West Africa. We previously described phenotypic differences between MAF and Mycobacterium tuberculosis (MTB) among 290 patients. In the present analysis, we compared 692 tuberculosis patients infected with the two most common lineages within the (MTB) complex found in the Gambia, namely MAF West African type 2 (39% prevalence) and Euro-American MTB (55% prevalence). We identified additional phenotypic differences between infections with these two organisms. MAF patients were more likely to be older and HIV infected. In addition, they had worse disease on chest X-ray, despite complaining of cough for an equal duration, and were more likely severely malnourished. In this cohort, the prevalence of MAF did not change significantly over a 7-year period. PMID- 20002177 TI - Adjuvants in tuberculosis vaccine development. AB - Tuberculosis remains a major public health problem around the world. Because the Mycobacterium bovis Bacilli-Calmette-Guerin (BCG) vaccine fails to protect adults from pulmonary tuberculosis, there is an urgent need for improved vaccine formulations. Unlike BCG, recombinant vaccines purified from bacterial expression vectors, as well as naked DNA, require an additional adjuvant. Recent improvements in our understanding of disease immunopathology, together with advances in biochemical and molecular techniques, have permitted the successful development of promising tuberculosis vaccine delivery and adjuvant combinations for human use. Here, we summarize the current state of adjuvant development and its impact on tuberculosis vaccine progress. PMID- 20002178 TI - Fungal diversity in oxygen-depleted regions of the Arabian Sea revealed by targeted environmental sequencing combined with cultivation. AB - In order to study fungal diversity in oxygen minimum zones of the Arabian Sea, we analyzed 1440 cloned small subunit rRNA gene (18S rRNA gene) sequences obtained from environmental samples using three different PCR primer sets. Restriction fragment length polymorphism (RFLP) analyses yielded 549 distinct RFLP patterns, 268 of which could be assigned to fungi (Dikarya and zygomycetes) after sequence analyses. The remaining 281 RFLP patterns represented a variety of nonfungal taxa, even when using putatively fungal-specific primers. A substantial number of fungal sequences were closely related to environmental sequences from a range of other anoxic marine habitats, but distantly related to known sequences of described fungi. Community similarity analyses suggested distinctively different structures of fungal communities from normoxic sites, seasonally anoxic sites and permanently anoxic sites, suggesting different adaptation strategies of fungal communities to prevailing oxygen conditions. Additionally, we obtained 26 fungal cultures from the study sites, most of which were closely related (>97% sequence similarity) to well-described Dikarya. This indicates that standard cultivation mainly produces more of what is already known. However, two of these cultures were highly divergent to known sequences and seem to represent novel fungal groups on high taxonomic levels. Interestingly, none of the cultured isolates is identical to any of the environmental sequences obtained. Our study demonstrates the importance of a multiple-primer approach combined with cultivation to obtain deeper insights into the true fungal diversity in environmental samples and to enable adequate intersample comparisons of fungal communities. PMID- 20002179 TI - Investigation of the faecal microbiota associated with canine chronic diarrhoea. AB - Diarrhoea is a common problem in dogs and can result in disturbance of the normal intestinal microbiota. However, little is known about the gastrointestinal microbiota of dogs with chronic diarrhoea and controlled canine studies of dietary management are scarce. The aims of this study were to investigate the predominant faecal microbiota of chronic diarrhoea dogs and to examine the effect(s) of a fibre blend on the canine faecal microbiota. A 3-week fibre supplementation feeding study was performed in nine chronic diarrhoea and eight control dogs. Atopobium cluster, Lactobacillus-Enterococcus group and Clostridium cluster XIV were the predominant bacterial groups in all dogs. Chronic diarrhoea dogs had significantly higher Bacteroides counts at baseline and significantly lower Atopobium cluster counts following fibre supplementation compared with control dogs. Atopobium cluster levels increased significantly in control dogs, while counts of sulphate-reducing bacteria decreased significantly and Clostridium clusters I and II counts increased significantly in chronic diarrhoea dogs during fibre supplementation. Microbial profiles (detected by denaturing gradient gel electrophoresis) demonstrated interindividual variation, with greater similarity seen between the chronic diarrhoea and control dogs' profiles after fibre supplementation compared with baseline. In conclusion, fibre supplementation induced changes in the canine faecal microbiota, with greater resemblance between the microbiota of chronic diarrhoea and control dogs after this dietary modulation. PMID- 20002180 TI - Stable carbon isotope fractionation by acetotrophic sulfur-reducing bacteria. AB - Acetate is the most important intermediate in anaerobic degradation of organic matter. The carbon isotope effects associated with the oxidation of acetate (epsilon(ac)) were examined for four acetotrophic sulfur reducers, Desulfuromonas acetoxidans, Desulfuromonas thiophila, Desulfurella acetivorans, and Hippea maritima. During the consumption of acetate and sulfur, acetate was enriched in (13)C by 11.5 and 11.2 per thousand in Desulfuromonas acetoxidans and Desulfuromonas thiophila, respectively. By contrast, isotope fractionation in D. acetivorans and H. maritima resulted in isotope enrichment factors of epsilon(ac)=-6.3 per thousand and -8.4 per thousand, respectively. These sulfur reducing bacteria all metabolize acetate via the tricarboxylic acid cycle, but have different mechanisms for the initial activation of acetate. In Desulfuromonas acetoxidans, acetyl-CoA is formed by succinyl-CoA : acetate-CoA transferase, and in D. acetivorans by acetate kinase and phosphate acetyltransferase. Hence, values of epsilon(ac) seem to be characteristic for the type of activation of acetate to acetyl-CoA in acetotrophic sulfur reducers. Summarizing epsilon(ac)-values in anaerobic acetotrophic microorganisms, it appears that isotope fractionation depends on the mechanism of acetate activation to acetyl-CoA, on the key enzyme of the acetate dissimilation pathway, and on the bioavailability of acetate, which all have to be considered when using delta(13)C of acetate in environmental samples for diagnosis of the involved microbial populations. PMID- 20002181 TI - Changes in fecal microbiota of healthy dogs administered amoxicillin. AB - The effect of oral amoxicillin treatment on fecal microbiota of seven healthy adult dogs was determined with a focus on the prevalence of bacterial antibiotic resistance and changes in predominant bacterial populations. After 4-7 days of exposure to amoxicillin, fecal Escherichia coli expressed resistance to multiple antibiotics when compared with the pre-exposure situation. Two weeks postexposure, the susceptibility pattern had returned to pre-exposure levels in most dogs. A shift in bacterial populations was confirmed by molecular fingerprinting of fecal bacterial populations using denaturing gradient gel electrophoresis (PCR-DGGE) of the 16S V3 rRNA gene region. Much of the variation in DGGE profiles could be attributed to dog-specific factors. However, permutation tests indicated that amoxicillin exposure significantly affected the DGGE profiles after controlling for the dog effect (P=0.02), and pre-exposure samples were clearly separated from postexposure samples. Sequence analysis of DGGE bands and real-time PCR quantification indicated that amoxicillin exposure caused a shift in the intestinal ecological balance toward a Gram-negative microbiota including resistant species in the family Enterobacteriaceae. PMID- 20002182 TI - Mechanisms that promote bacterial fitness in fungal-affected soil microhabitats. AB - Soil represents a very heterogeneous environment for its microbiota. Among the soil inhabitants, bacteria and fungi are important organisms as they are involved in key biogeochemical cycling processes. A main energy source driving the system is formed by plants through the provision of plant-fixed (reduced) carbon to the soil, whereas soil nitrogen and phosphorus may move from the soil back to the plant. The carbonaceous compounds released form the key energy and nutrient sources for the soil microbiota. In the grossly carbon-limited soil, the emergence of plant roots and the formation of their associated mycorrhizae thus create nutritional hot spots for soil-dwelling bacteria. As there is natural (fitness) selection on bacteria in the soil, those bacteria that are best able to benefit from the hot spots have probably been selected. The purpose of this review is to examine the interactions of bacteria with soil fungi in these hot spots and to highlight the key mechanisms involved in the selection of fungal responsive bacteria. Salient bacterial mechanisms that are involved in these interactions have emerged from this examination. Thus, the efficient acquisition for specific released nutrients, the presence of type-III secretion systems and the capacity of flagellar movement and to form a biofilm are pinpointed as key aspects of bacterial life in the mycosphere. The possible involvement of functions present on plasmid-borne genes is also interrogated. PMID- 20002183 TI - Vibrio owensii sp. nov., isolated from cultured crustaceans in Australia. AB - Two bacterial strains (DY05(T) and 47666-1) were isolated in Queensland, Australia, from diseased cultured crustaceans Panulirus ornatus and Penaeus monodon, respectively. On the basis of 16S rRNA gene sequence identity, the strains were shown to belong to the Harveyi clade of the genus Vibrio. Multilocus sequence analysis using five housekeeping genes (rpoA, pyrH, topA, ftsZ and mreB) showed that the strains form a monophyletic group with 94.4% concatenated sequence identity to the closest species. DNA-DNA hybridization experiments showed that strains DY05(T) and 47666-1 had 76% DNA similarity to each other, but <70% to their closest neighbours Vibrio harveyi LMG 4044(T) (< or =55%), Vibrio campbellii LMG 11216(T) (< or =52%) and Vibrio rotiferianus LMG 21460(T) (< or =46%). Strains DY05(T) and 47666-1 could be differentiated from their relatives on the basis of several phenotypic characteristics. The major fatty acids were C(15:0) iso 2-OH and/or C(16:1)omega7, C(16:0), C(18:1)omega7 and C(14:0). Based on the polyphasic evidence presented here, it can be concluded that strains DY05(T) and 47666-1 belong to the same novel species of the genus Vibrio, for which the name Vibrio owensii sp. nov. is proposed. The type strain is DY05(T) (=JCM 16517(T)=ACM 5300(T)). PMID- 20002184 TI - The role of Sov protein in the secretion of gingipain protease virulence factors of Porphyromonas gingivalis. AB - Porphyromonas gingivalis transports Arg-gingipains and Lys-gingipain across the outer membrane via an unknown pathway. Recently, we found that the sov gene of P. gingivalis W83 was required for this step. In the present study, we characterized the Sov protein. We constructed a P. gingivalis strain that expresses histidine tagged Sov instead of Sov. Subcellular fractionations and a histidine-tag pulldown experiment showed that histidine-tagged Sov was present in an outer membrane fraction. Furthermore, antiserum raised against the terminal regions of Sov obstructed the secretion of Arg-gingipains from wild-type W83 cells. A deletion study showed that the region from Phe2495 to the C-terminus Gln2499 of Sov is essential for gingipain secretion. Anti-histidine-tag immunoglobulins interfered with the secretion of Arg-gingipains by P. gingivalis cells that expressed histidine-tagged Sov. In conclusion, we found that Sov is an outer membrane protein participating in the secretion of gingipains and that the C terminal region of Sov is exposed to the extracellular milieu and involved in the modulation of Sov function. PMID- 20002185 TI - Activity of Bacillus thuringiensis hybrid protein against a lepidopteran and a coleopteran pest. AB - The use of Cry proteins from Bacillus thuringiensis are an important strategy for biological control. Recently it has been demonstrated that Cry hybrid proteins (by domain swapping) resulted in improved toxicities in comparison with parental proteins. Here, an SN1917 hybrid toxin was constructed and tested against Colombian pest insects Tecia solanivora (Lepidoptera: Gelechiidae), a severe potato pest, and Hypothenemus hampei (Coleoptera: Scolytidae), which attacks coffee crops. The SN1917 protoxin had a concentration causing 50% mortality (LC(50)) of 392 ng cm(-2), and SN1917 toxin showed an LC(50) of 201 ng cm(-2) against T. solanivora first instar larvae. The two parental toxins (Cry1B and Cry1I) used to make this new hybrid protein caused a mortality of 60% and 52%, respectively. Unfortunately, H. hampei first instar larvae proved to be resistant to the toxin. We conclude that SN1917 is an option for biological control and resistance management of T. solanivora. Implications for H. hampei are discussed. PMID- 20002186 TI - Characterization of carotenoid biosynthetic genes in the ascomycete Gibberella zeae. AB - Carotenoids are a structurally diverse class of terpenoid pigments that are synthesized by many microorganisms and plants. In this study, we identified five putative carotenoid biosynthetic genes from the ascomycete Gibberella zeae (GzCarB, GzCarO, GzCarRA, GzCarT, and GzCarX). HPLC showed that the fungus produces two carotenoids: neurosporaxanthin and torulene. We deleted the five genes individually to determine their functions. GzCarB, GzCarRA, and GzCarT were required for neurosporaxanthin biosynthesis, but the deletion of GzCarX or GzCarO (DeltagzcarX or DeltagzcarO) failed to alter the production of neurosporaxanthin or torulene. DeltagzcarRA and DeltagzcarB did not produce neurosporaxanthin or torulene. DeltagzcarB led to the accumulation of phytoene, which is an intermediate in carotenoid biosynthesis, but DeltagzcarRA did not. DeltagzcarT produced torulene but not neurosporaxanthin. Based on these functional studies and similarities to carotenoid biosynthesis genes in other fungi, we deduced the functions of the three genes and propose the carotenoid biosynthetic pathway of G. zeae. PMID- 20002187 TI - Live and lyophilized Lactobacillus species elicit differential immunomodulatory effects on immune cells. AB - To study the mechanism of action of the lactobacilli, splenocytes were incubated with lactobacilli. We compared the ability of live and lyophilized Lactobacillus casei, Lactobacillus rhamnosus GG and Lactobacillus delbrueckii ssp. bulgaricus to modulate the production of interleukin 12p40 (IL-12p40), tumor necrosis factor alpha and IL-10 by splenocytes from C57BL/6 and BALB/c mice. Blocking contact between lactobacilli and immune cells abrogated all cytokine production. Toll like receptor 2 (TLR2) was partially responsible, but not TLR4 or TLR9, for the induction of cytokine production in splenocytes. All cytokine production declined to basal levels when bacterial phagocytosis was inhibited. This shows that lactobacilli stimulation of cytokine production in splenocytes requires the process of phagocytosis and engagement of TLR2, but not TLR4 or TLR9. PMID- 20002188 TI - Salicylidene acylhydrazide-mediated inhibition of type III secretion system-1 in Salmonella enterica serovar Typhimurium is associated with iron restriction and can be reversed by free iron. AB - Salmonella enterica serovar Typhimurium is an animal and zoonotic pathogen of worldwide importance. Intestinal colonization, induction of enteritis and systemic translocation by this bacterium requires type III protein secretion. Strategies that target this process have the potential to control infection, pathology and transmission. We defined the global transcriptional response of S. Typhimurium to INP0403, a member of a family of salicylidene acylhydrazides that inhibit type III secretion (T3S). INP0403 treatment was associated with reduced transcription of genes involved in T3S, but also increased transcription of genes associated with iron acquisition. We show that INP0403 restricts iron availability to Salmonella, and that inhibition of T3S system-1 by INP0403 is, at least in part, reversible by exogenous iron and independent of the iron response regulator Fur. PMID- 20002189 TI - Biogenesis of Yersinia pestis PsaA in recombinant attenuated Salmonella Typhimurium vaccine (RASV) strain. AB - Yersinia pestis PsaA is an adhesin important for the establishment of bacterial infection. PsaA synthesis requires the products of the psaEFABC genes. Here, by prediction analysis, we identified a PsaA signal sequence with two signal peptidase (SPase) cleavage sites, type-I and type-II (SPase-I and SPase-II). By Edman degradation and site-directed mutagenesis, the precise site for one of these Spase-I PsaA cleavage sites was located between alanine and serine at positions 31 and 32, respectively. Yersinia pestis psaA expression and the role of the PsaB and PsaC proteins were evaluated in recombinant attenuated Salmonella Typhimurium vaccine strains. PsaA was detected in total extracts as a major 15 kDa (mature) and 18-kDa (unprocessed) protein bands. PsaA synthesis was not altered by a DeltaA31-DeltaS32 double-deletion mutation. In contrast, the synthesis of PsaA (DeltaA31-DeltaS32) in Y. pestis and delivery to the supernatant was decreased. Otherwise, substitution of the amino acid cysteine at position 26 by valine involved in the SPase-II cleavage site did not show any effect on the secretion of PsaA in Salmonella and Yersinia. These results help clarify the secretion pathway of PsaA for the possible development of vaccines against Y. pestis. PMID- 20002190 TI - Catalase-peroxidase activity is decreased in a Caulobacter crescentus rho mutant. AB - A Caulobacter crescentus rho::Tn5 mutant strain presenting a partially functional transcription termination factor Rho is highly sensitive to hydrogen peroxide in both exponential and stationary phases. The mutant was shown to be permanently under oxidative stress, based on fluorophore oxidation, and also to be sensitive to tert-butyl hydroperoxide and paraquat. However, the results showed that the activities of superoxide dismutases CuZnSOD and FeSOD and the alkylhydroperoxide reductase ahpC mRNA levels in the rho mutant were comparable to the wild-type control in the exponential and stationary phases. In contrast, the KatG catalase activity of the rho mutant strain was drastically decreased and did not show the expected increase in the stationary phase compared with the exponential phase. Transcription of the katG gene was increased in the rho mutant and the levels of the immunoreactive KatG protein do not differ considerably compared with the wild type in the stationary phase, suggesting that KatG activity is affected in a translational or a post-translational step. PMID- 20002191 TI - Purification and characterization of vanillin dehydrogenases from alkaliphile Micrococcus sp. TA1 and neutrophile Burkholderia cepacia TM1. AB - Vanillin dehydrogenases (VDHs) were purified and characterized from two bacterial strains that have different pH dependencies for growth. The alkaliphile Micrococcus sp. TA1, isolated from an alkaline spa, can grow on several aromatic compounds such as ferulic acid, vanillin, vanillic acid, and protocatechuic acid under alkaline conditions. The neutrophile Burkholderia cepacia TM1, which was isolated previously, also grew on the above-mentioned compounds because they functioned as the sole carbon source under neutral conditions. Purified VDHs showed activities toward some aromatic aldehydes. These enzymes have the same subunit molecular mass of about 57 kDa as determined by sodium dodecyl sulfate polyacrylamide gel electrophoresis, but differed in some of their observed properties. Native molecular masses also differed between the purified enzymes. These were 250 kDa for the enzyme from alkaliphilic strain TA1 and 110 kDa for that from neutrophilic strain TM1, as determined by gel filtration. The enzyme from strain TA1 required NADP(+) as a coenzyme for its activity, but that from strain TM1 required NAD(+). These results are important because this is the first report of an alkaliphilic bacterium consuming lignin monomers. PMID- 20002192 TI - Zoosporic plant pathogens produce bacterial autoinducer-2 that affects Vibrio harveyi quorum sensing. AB - The frequent coisolation of bacteria with Phytophthora and Pythium species suggests possible interspecies communication. Zoospore-free fluids (ZFF) from bacteria-free and nutrient-depleted zoospore suspensions were examined to investigate the production of autoinducer-2 (AI-2), a bacterial interspecies signal molecule, by zoosporic oomycetes. ZFF from Phytophthora nicotianae, Phytophthora sojae, and Pythium aphanidermatum triggered luminescence of the Vibrio harve7yi AI-2 reporter, indicating the presence of AI-2 in zoospore extracellular products and the potential of cross-kingdom communication between oomycetes and bacteria. The production of AI-2 by zoospores was confirmed by chemical assays. These results provide a new insight into the physiology and ecology of oomycetes. PMID- 20002193 TI - Identification of amino acids required for receptor binding and toxicity of the Bacillus sphaericus binary toxin. AB - Bacillus sphaericus produces a mosquito-larvicidal binary toxin composed of BinB and BinA subunits. BinA is important for toxicity, whereas BinB acts as a specific receptor-binding component. To study the functional significance of two regions that are only present in BinB, four block mutations and two single mutations were initially introduced: (111)YLD(113)-->(111)AAA(113), (115)NNH(117) ->(115)AAA(117), (143)GEQ(145)-->(143)AAA(145), (147)FQFY(150)-->(147)AAAA(150), N114A and F146A. Only the replacements at (147)FQFY(150) resulted in a total loss of toxicity to Culex quinquefasciatus larvae. Further single alanine substitutions in this region, F147A, Q148A, F149A and Y150A, were introduced to identify residues playing a critical role in mosquito-larvicidal activity. Larvicidal activity assays revealed that only F149A and Y150A mutants exhibited a total loss of toxicity. The in vitro interaction assays demonstrated that all BinB mutants are able to interact with BinA. Immunohistochemistry analysis revealed that only the Y150A mutant was unable to bind to the larval midgut, suggesting an important role of this residue in receptor binding of the BinB subunit. Conservative aromatic substitutions at F149 and Y150 resulted in full recovery of larvicidal activity, indicating that the aromaticity of F149 and Y150 is a key determinant of larvicidal activity, possibly playing a key role in the membrane interaction and receptor binding. PMID- 20002194 TI - Scl1, the multifunctional adhesin of group A Streptococcus, selectively binds cellular fibronectin and laminin, and mediates pathogen internalization by human cells. AB - The streptococcal collagen-like protein-1, Scl1, is widely expressed by the well recognized human pathogen group A Streptococcus (GAS). Screening of human ligands for binding to recombinant Scl1 identified cellular fibronectin and laminin as binding partners. Both ligands interacted with the globular domain of Scl1, which is also able to bind the low-density lipoprotein. Native Scl1 mediated GAS adherence to ligand-coated glass cover slips and promoted GAS internalization into HEp-2 cells. This work identifies new ligands of the Scl1 protein that are known to be important in GAS pathogenesis and suggests a novel ligand-switching mechanism between blood and tissue environments, thereby facilitating host colonization and GAS dissemination. PMID- 20002195 TI - Atorvastatin-induced cell toxicity in yeast is linked to disruption of protein isoprenylation. AB - Statins, used to treat hypercholesterolemia, are one of the most frequently prescribed drug classes in the developed world. However, a significant proportion of users suffer symptoms of myotoxicity, and currently, the molecular mechanisms underlying myotoxicity remain ambiguous. In this study, Saccharomyces cerevisiae was exploited as a model system to gain further insight into the molecular mechanisms of atorvastatin toxicity. Atorvastatin-treated yeast cells display marked morphological deformities, have reduced cell viability and are highly vulnerable to perturbed mitochondrial function. Supplementation assays of atorvastatin-treated cells reveal that both loss of viability and mitochondrial dysfunction occur as a consequence of perturbation of the sterol synthesis pathway. This was further investigated by supplementing statin-treated cells with various metabolites of the sterol synthesis pathway that are believed to be essential for cell function. Ergosterol, coenzyme Q and a heme precursor were all ineffective in the prevention of statin-induced mitochondrial disruption and cell death. However, the addition of geranylgeranyl pyrophosphate and farnesyl pyrophosphate significantly restored cell viability, although these did not overcome petite induction. This highlights the pleiotropic nature of statin toxicity, but has established protein prenylation disruption as one of the principal mechanisms underlying statin-induced cell death in yeast. PMID- 20002196 TI - Detection of a homotetrameric structure and protein-protein interactions of Paracoccidioides brasiliensis formamidase lead to new functional insights. AB - Paracoccidioides brasiliensis causes paracoccidioidomycosis, a systemic mycosis in Latin America. Formamidases hydrolyze formamide, putatively plays a role in fungal nitrogen metabolism. An abundant 45-kDa protein was identified as the P. brasiliensis formamidase. In this study, recombinant formamidase was overexpressed in bacteria and a polyclonal antibody to this protein was produced. We identified a 180-kDa protein species reactive to the antibody produced in mice against the P. brasiliensis recombinant purified formamidase of 45 kDa. The 180 kDa purified protein yielded a heat-denatured species of 45 kDa. Both protein species of 180 and 45 kDa were identified as formamidase by peptide mass fingerprinting using MS. The identical mass spectra generated by the 180 and the 45-kDa protein species indicated that the fungal formamidase is most likely homotetrameric in its native conformation. Furthermore, the purified formamidase migrated as a protein of 191 kDa in native polyacrylamide gel electrophoresis, thus revealing that the enzyme forms a homotetrameric structure in its native state. This enzyme is present in the fungus cytoplasm and the cell wall. Use of a yeast two-hybrid system revealed cell wall membrane proteins, in addition to cytosolic proteins interacting with formamidase. These data provide new insights into formamidase structure as well as potential roles for formamidase and its interaction partners in nitrogen metabolism. PMID- 20002197 TI - Role of outer-membrane cytochromes MtrC and OmcA in the biomineralization of ferrihydrite by Shewanella oneidensis MR-1. AB - In an effort to improve the understanding of electron transfer mechanisms at the microbe-mineral interface, Shewanella oneidensis MR-1 mutants with in-frame deletions of outer-membrane cytochromes (OMCs), MtrC and OmcA, were characterized for the ability to reduce ferrihydrite (FH) using a suite of microscopic, spectroscopic, and biochemical techniques. Analysis of purified recombinant proteins demonstrated that both cytochromes undergo rapid electron exchange with FH in vitro with MtrC displaying faster transfer rates than OmcA. Immunomicroscopy with cytochrome-specific antibodies revealed that MtrC co localizes with iron solids on the cell surface while OmcA exhibits a more diffuse distribution over the cell surface. After 3-day incubation of MR-1 with FH, pronounced reductive transformation mineral products were visible by electron microscopy. Upon further incubation, the predominant phases identified were ferrous phosphates including vivianite [Fe(3)(PO(4))(2)x8H(2)O] and a switzerite like phase [Mn(3),Fe(3)(PO(4))(2)x7H(2)O] that were heavily colonized by MR-1 cells with surface-exposed outer-membrane cytochromes. In the absence of both MtrC and OmcA, the cells ability to reduce FH was significantly hindered and no mineral transformation products were detected. Collectively, these results highlight the importance of the outer-membrane cytochromes in the reductive transformation of FH and support a role for direct electron transfer from the OMCs at the cell surface to the mineral. PMID- 20002198 TI - The polypill in the primary prevention of cardiovascular disease. AB - Coronary heart disease (CHD) and stroke are the most common causes of death worldwide. The polypill, a multi-component tablet or capsule designed to reduce several cardiovascular causal risk factors simultaneously, has the potential to reduce the incidence of heart attacks and stroke by about 80%, the precise reduction depending on the formulation of the polypill and adherence to preventive treatment. The full public health impact can only be realized if the polypill is used to prevent first cardiovascular disease events, because most heart attacks and strokes are first events. A simple and effective method of selecting who should be offered the polypill is needed. Even though serum cholesterol and blood pressure are important causes of CHD and stroke and lowering them has a large preventive effect (a 1.8 mmol/L reduction in LDL cholesterol reduces the incidence of CHD events by about 60% and a 10 mmHg reduction in diastolic blood pressure also reduces the incidence of stroke by about 60%), they are poor screening tests. They add little to age in discriminating between individuals who will and will not have a CHD event or stroke. Including them in a risk assessment algorithm needlessly complicates the screening process and tends to medicalize the preventive strategy. Age alone is effective and a simpler means of selecting people for preventive treatment using the polypill. PMID- 20002199 TI - Distinct thermal migration behaviors in response to different thermal gradients in Caenorhabditis elegans. AB - The nematode Caenorhabditis elegans exhibits a complex behavior called thermotaxis in response to temperature. This behavior is defined as a form of associative learning, in which temperature pairs with the presence or absence of food. Different interpretations have been drawn from the diverse results obtained by several groups, mainly because of the application of different methodologies for the analysis of thermotaxis. To clarify the discrepancies in behavioral observations and subsequent interpretations by different laboratories, we attempted to systematize several parameters to observe thermotaxis behavior as originally defined by Hedgecock and Russell in 1975. In this study, we show clearly how C. elegans can show a conditioned migration toward colder or warmer areas on a thermal gradient, given certain criteria necessary for the observation of thermotaxis. We thus propose to distinguish thermotaxis from other temperature related behaviors, such as the warm avoidance response displayed at temperature gradients of 1 degrees C/cm and steeper. PMID- 20002200 TI - COMT but not serotonin-related genes modulates the influence of childhood abuse on anger traits. AB - Anger-related traits are regulated by genes as well as early environmental factors. Both childhood maltreatment and genes underlie vulnerability to suicidal behaviors, possibly by affecting the constitution of intermediate phenotypes such as anger traits. The aim of this study was to test the interaction between nine candidate genes and childhood maltreatment in modulating anger-related traits in 875 adult suicide attempters. The State-Trait Anger Expression Inventory and the Childhood Trauma Questionnaire were used to examine anger traits and traumatic childhood experiences, respectively. The functional polymorphism of the catecholamine-O-methyl-transferase (COMT) gene Val158Met significantly modulated the association between sexual abuse and anger-trait level (P = 0.001). In the presence of sexual abuse, individuals carrying the Val high-activity allele displayed greater disposition toward anger than individuals homozygous for the Met allele (P = 0.0003). Notably, none of the serotonin-related genes influenced the effect of childhood abuse on anger traits. The results of the present study suggest that anger-trait level is influenced by the interaction between childhood abuse and functional polymorphism in the COMT gene. This study was carried out in a population with a high frequency of childhood abuse and a high disposition toward anger, and replication in healthy subjects is needed. PMID- 20002201 TI - Individual vulnerability to escalated aggressive behavior by a low dose of alcohol: decreased serotonin receptor mRNA in the prefrontal cortex of male mice. AB - Low to moderate doses of alcohol consumption induce heightened aggressive behavior in some, but not all individuals. Individual vulnerability for this nonadaptive behavior may be determined by an interaction of genetic and environmental factors with the sensitivity of alcohol's effects on brain and behavior. We used a previously established protocol for alcohol oral self administration and characterized alcohol-heightened aggressive (AHA) mice as compared with alcohol non-heightened (ANA) counterparts. A week later, we quantified mRNA steady state levels of several candidate genes in the serotonin [5-hydroxytryptamine (5-HT)] system in different brain areas. We report a regionally selective and significant reduction of all 5-HT receptor subtype transcripts, except for 5-HT(3), in the prefrontal cortex of AHA mice. Comparable gene expression profile was previously observed in aggressive mice induced by social isolation or by an anabolic androgenic steroid. Additional change in the 5 HT(1B) receptor transcripts was seen in the amygdala and hypothalamus of AHA mice. In both these areas, 5-HT(1B) mRNA was elevated when compared with ANA mice. In the hypothalamus, AHA mice also showed increased transcripts for 5 HT(2A) receptor. In the midbrain, 5-HT synthetic enzyme, 5-HT transporter and 5 HT receptors mRNA levels were similar between groups. Our results emphasize a role for postsynaptic over presynaptic 5-HT receptors in mice which showed escalated aggression after the consumption of a moderate dose of alcohol. This gene expression profile of 5-HT neurotransmission components in the brain of mice may suggest a vulnerability trait for alcohol-heightened aggression. PMID- 20002203 TI - Isoform-specific effects of apolipoprotein E on cognitive performance in targeted replacement mice overexpressing human APP. AB - The epsilon 4 allele of apolipoprotein E (apoE4) is the predominant genetic risk factor for late-onset Alzheimer's disease (AD) and is also implicated in cognitive deficits associated with normal aging. The biological mechanisms by which APOE genotype affects cognitive processes or AD pathogenesis remain unclear, but interactions of apoE with amyloid beta peptide (A beta) are thought to play an important role in mediating apoE's isoform-specific effects on brain function. Here, we investigated the potential isoform-dependent effects of apoE on behavioral and cognitive performance in human apoE3 and apoE4 targeted replacement (TR) mice that also overexpress the human amyloid precursor protein (APP). Beginning at 6-7 months of age, female APP-Yac/apoE3-TR ('poE3') and APP Yac/apoE4-TR ('poE4') mice were tested on a battery of tests to evaluate basic sensorimotor functioning, spatial working memory, spatial recognition, episodic like memory and attentional processing. Compared with apoE3 mice, a generalized reduction in locomotor activity was observed in apoE4 mice. Moderate, but significant, cognitive impairments were also detected in apoE4 mice in the novel object-location preference task, the contextual fear conditioning test, and a two choice visual discrimination/detection test, however spontaneous alternation performance in the Y-maze was spared. These results offer additional support for the negative impact of apoE4 on both memory and attention and further suggest that APP-Yac/apoE-TR mice provide a novel and useful model for investigating the role of apoE in mediating susceptibility to cognitive decline. PMID- 20002202 TI - MitoPark mice mirror the slow progression of key symptoms and L-DOPA response in Parkinson's disease. AB - The MitoPark mouse, in which the mitochondrial transcription factor Tfam is selectively removed in midbrain dopamine (DA) neurons, is a genetic model for Parkinson's disease (PD) that replicates the slow and progressive development of key symptoms. To further validate this model, we have extended both behavioral and biochemical analyses in these animals. We found that vertical movements decline earlier and faster than horizontal movements, possibly modeling the early occurrence of axial, postural instability in PD. L-DOPA induces different locomotor responses depending on the age: in young MitoPark mice the L-DOPA induced motor activation is small; middle-aged MitoPark mice respond in a dose dependent manner to L-DOPA, whereas aged MitoPark mice display a double-peaked locomotor response to a high dose of L-DOPA that includes an intermittent period of very low motor activity, similar to the 'on-off' phenomenon in PD. To correlate behavior with biochemical data, we analyzed monoamine levels in three different brain areas that are highly innervated by the DA system: striatum, anterior cortex and olfactory bulb. DA levels declined earlier and faster in striatum than in cortex; only at the latest time-point analyzed, DA levels were found to be significantly lower than control levels in the olfactory bulb. Interestingly, the ratio between homovanillic acid (HVA) and DA differed between regions over time. In striatum and olfactory bulb, the ratio increased steeply indicating increased DA turnover. In contrast, the ratio decreased over time in cortex, revealing important differences between DA cells in substantia nigra and the ventral tegmental area. PMID- 20002204 TI - Microbiology and geochemistry of Little Hot Creek, a hot spring environment in the Long Valley Caldera. AB - A culture-independent community census was combined with chemical and thermodynamic analyses of three springs located within the Long Valley Caldera, Little Hot Creek (LHC) 1, 3, and 4. All three springs were approximately 80 degrees C, circumneutral, apparently anaerobic and had similar water chemistries. 16S rRNA gene libraries constructed from DNA isolated from spring sediment revealed moderately diverse but highly novel microbial communities. Over half of the phylotypes could not be grouped into known taxonomic classes. Bacterial libraries from LHC1 and LHC3 were predominantly species within the phyla Aquificae and Thermodesulfobacteria, while those from LHC4 were dominated by candidate phyla, including OP1 and OP9. Archaeal libraries from LHC3 contained large numbers of Archaeoglobales and Desulfurococcales, while LHC1 and LHC4 were dominated by Crenarchaeota unaffiliated with known orders. The heterogeneity in microbial populations could not easily be attributed to measurable differences in water chemistry, but may be determined by availability of trace amounts of oxygen to the spring sediments. Thermodynamic modeling predicted the most favorable reactions to be sulfur and nitrate respirations, yielding 40-70 kJ mol(-1) e(-) transferred; however, levels of oxygen at or below our detection limit could result in aerobic respirations yielding up to 100 kJ mol(-1) e(-) transferred. Important electron donors are predicted to be H(2), H(2)S, S(0), Fe(2+) and CH(4), all of which yield similar energies when coupled to a given electron acceptor. The results indicate that springs associated with the Long Valley Caldera contain microbial populations that show some similarities both to springs in Yellowstone and springs in the Great Basin. PMID- 20002205 TI - Implant-retained thumb prosthesis with anti-rotational attachment for a geriatric patient. AB - This report presents the use of a dental implant with an anti-rotational attachment for the retention of a thumb prosthesis. A retention system was manufactured with an attachment (UCLA) screwed into the implant with a two-bar system that was cast in metallic silver palladium. A substructure made from heat cured acrylic resin was joined to the retention system by clips to join the thumb to the finger (bar clip) in the cast with implant rejoinder. The silicone material, Silastic-MDX 44210, was used to achieve function and aesthetics. Following osseointegration, no skin problems were observed. Whilst the implant retained digital prosthesis presented some motor limitations, it allowed the patient to return to entertainment and achieve social conviviality. PMID- 20002206 TI - Methods to derive the differential equation of the free surface boundary. PMID- 20002207 TI - Transient divergent flow and transport in an infinite anisotropic porous formation. AB - When seeking to predict plume geometry resulting from fluid injection through partially penetrating wells, it is common to assume a steady-state spherically diverging flow field. In reality, the flow field is transient. The steady-flow assumption is likely to cause overestimation of injection plume radius since the accommodation of fluid by increases in porosity and fluid density is ignored. In this paper, a transient solution is developed, resulting in a nonlinear ordinary differential equation expressing plume radius as a function of time. It is shown that the problem can be fully described by one type curve. A critical time, t(c), is identified at which the percentage error of the steady-state flow solution compared to the fully dynamic problem is less than 1%. Only for large injection rates and low permeabilities, does t(c) become greater than 1 h. Nevertheless, an improved approximate solution is obtained by a simple linearization procedure. The critical time, t(c) for the new approximate solution is 0.3% of that required for the steady-state flow solution. PMID- 20002208 TI - Multiple well-shutdown tests and site-scale flow simulation in fractured rocks. AB - A new method was developed for conducting aquifer tests in fractured-rock flow systems that have a pump-and-treat (P&T) operation for containing and removing groundwater contaminants. The method involves temporary shutdown of individual pumps in wells of the P&T system. Conducting aquifer tests in this manner has several advantages, including (1) no additional contaminated water is withdrawn, and (2) hydraulic containment of contaminants remains largely intact because pumping continues at most wells. The well-shutdown test method was applied at the former Naval Air Warfare Center (NAWC), West Trenton, New Jersey, where a P&T operation is designed to contain and remove trichloroethene and its daughter products in the dipping fractured sedimentary rocks underlying the site. The detailed site-scale subsurface geologic stratigraphy, a three-dimensional MODFLOW model, and inverse methods in UCODE_2005 were used to analyze the shutdown tests. In the model, a deterministic method was used for representing the highly heterogeneous hydraulic conductivity distribution and simulations were conducted using an equivalent porous media method. This approach was very successful for simulating the shutdown tests, contrary to a common perception that flow in fractured rocks must be simulated using a stochastic or discrete fracture representation of heterogeneity. Use of inverse methods to simultaneously calibrate the model to the multiple shutdown tests was integral to the effectiveness of the approach. PMID- 20002209 TI - Sustaining a positive body image in adolescence: an assets-based analysis. AB - The increasing impetus to provide an effective response to childhood obesity has generated a corresponding concern that weight management interventions may lead to emotional problems among young people, notably in relation to the development of a negative body image. However, currently the processes and factors that contribute to the acquisition of body image among young people is poorly understood. Drawing on salutogenic theory, this paper employs an assets-based approach that focuses on health promoting and protective factors to identify how young people may create or sustain positive body images. Secondary data analysis was undertaken from the WHO Health Behaviour in School Aged Children Study. During the spring term of 2002, information was collected from 6425 English adolescents aged 11-15 using a self-administered questionnaire. The data were analysed using stepwise multinomial logistic regression to determine which factors were associated with positive body image; a total of 2898 students were included in the final analysis. Adolescents who self-identified as having a positive body image were more likely to report ease of talking with a father figure, feeling intelligent, perceiving that their family were well off and a belief that their teachers were interested in them as people. Body Mass Index, age, gender and living within a household containing a father were also significant predictors of body image. The discussion provides an exploration of how the construction of young people's emotional health is in part linked with the attainment of a secure and positive body image. The implications for health promotion and educational programmes are then examined. By considering the assets, which support or sustain a positive body image during adolescence, obesity prevention programmes could be better tailored to meet the needs of young people. In the future, a salutogenic curriculum might provide an alternative to unsustainable levels of deficit led, targeted programmes. PMID- 20002210 TI - Dendritic cells treated with resveratrol during differentiation from monocytes gain substantial tolerogenic properties upon activation. AB - Resveratrol is a polyphenol that acts on multiple molecular targets important for cell differentiation and activation. Dendritic cells (DCs) are a functionally diverse cell type and represent the most potent antigen-presenting cells of the immune system. In this study, we investigated resveratrol-induced effects on DCs during their differentiation and maturation. Our results show that resveratrol induces DC-associated tolerance, particularly when applied during DC differentiation. Costimulatory molecules CD40, CD80 and CD86 were down-regulated, as was the expression of major histocompatibility complex (MHC) class II molecules. Surface expression of inhibitory immunoglobulin-like transcript 3 (ILT3) and ILT4 molecules was induced, while human leucocyte antigen (HLA)-G expression was not affected. Resveratrol-treated DCs lost the ability to produce interleukin (IL)-12p70 after activation, but had an increased ability to produce IL-10. Such DCs were poor stimulators of allogeneic T cells and had lowered ability to induce CD4(+) T-cell migration. Furthermore, treated cells were able to generate allogeneic IL-10-secreting T cells, but were not competent in inducing FoxP3 expression These tolerogenic effects are probably associated with the effect of resveratrol on multiple molecular targets through which it interferes with DC differentiation and nuclear factor (NF)-kappaB translocation. Our data provide new insights into the molecular and functional mechanisms of the tolerogenic effects that resveratrol exerts on DCs. PMID- 20002211 TI - Cytoskeletal proteins bound to heat-shock protein 70 may elicit resistance to simian immunodeficiency virus infection of CD4(+) T cells. AB - This study is based on the evidence that immunization of macaques with human CD4(+) T cells elicits prevention of simian immunodeficiency virus (SIV) infection. We hypothesized that heat-shock protein 70 (HSP70) isolated from CD4(+) T cells may act as a chaperone and carry the protective host proteins. Two moieties of HSP70 were affinity-purified from human CD4(+) T cells; an ADP preparation with HSP70-bound proteins (ADP-HSP) and an ATP control preparation. Immunization of rhesus macaques with these preparations showed significant inhibition of SIVmac251 infectivity ex vivo in CD4(+) T cells only with the ADP HSP (P = 0.01). Proteomic analysis identified three cytoskeletal elements, cofilin, profilin and gamma-actin, exclusively in the ADP-HSP preparation. Investigation of the mechanism of prevention of SIV replication suggests that antibodies to the cytoskeletal proteins may inhibit actin depolymerization and facilitate viral degradation by the innate antiviral APOBEC3G. As cytoskeletal proteins are critical in the formation of virological and immunological synapses, finding specific antibodies and anti-SIV/human immunodeficiency virus (HIV) factors suggests a novel insight into HIV-1 immunopathogenesis. PMID- 20002212 TI - Extensive major histocompatibility complex class I binding promiscuity for Mycobacterium tuberculosis TB10.4 peptides and immune dominance of human leucocyte antigen (HLA)-B*0702 and HLA-B*0801 alleles in TB10.4 CD8 T-cell responses. AB - The molecular definition of major histocompatibility complex (MHC) class I presented CD8(+) T-cell epitopes from clinically relevant Mycobacterium tuberculosis (Mtb) target proteins will aid in the rational design of T-cell based diagnostics of tuberculosis (TB) and the measurement of TB vaccine-take. We used an epitope discovery system, based on recombinant MHC class I molecules that cover the most frequent Caucasian alleles [human leucocyte antigen (HLA)-A*0101, A*0201, A*0301, A*1101, A*2402, B*0702, B*0801 and B*1501], to identify MHC class I-binding peptides from overlapping 9-mer peptides representing the Mtb protein TB10.4. A total of 33 MHC class I-binding epitopes were identified, spread across the entire amino acid sequence, with some clustering at the N- and C-termini of the protein. Binding of individual peptides or closely related peptide species to different MHC class I alleles was frequently observed. For instance, the common motif of xIMYNYPAMx bound to six of eight alleles. Affinity (50% effective dose) and off-rate (half life) analysis of candidate Mtb peptides will help to define the conditions for CD8(+) T-cell interaction with their nominal MHC class I peptide ligands. Subsequent construction of tetramers allowed us to confirm the recognition of some of the epitopes by CD8(+) T cells from patients with active pulmonary TB. HLA-B alleles served as the dominant MHC class I restricting molecules for anti-Mtb TB10.4-specific CD8(+) T-cell responses measured in CD8(+) T cells from patients with pulmonary TB. PMID- 20002213 TI - Extensive gene duplication of acetylcholinesterase associated with organophosphate resistance in the two-spotted spider mite. AB - Monocrotophos-resistant two-spotted spider mites (TSSMs), Tetranychus urticae, are known to possess three mutations on the acetylcholinesterase (AChE) gene (Tuace) that are involved in target site insensitivity. Cross-strain comparison of three strains (highly resistant AD, moderately resistant PyriF and susceptible UD strains) revealed that resistant strains have relatively more Tuace copies than the UD strain and that the levels of transcript were directly proportional to copy numbers. AChEs from the AD and PyriF strains had similar V(max) values to those of AChE from the UD strain but increased K(m) and reduced k(cat) constants, suggesting that the mutated, resistant form of AChE may carry a fitness cost. Relative copy numbers of Tuace in field populations varied from 2.4 to 6.1, correlating well with their levels of resistance (r(2)= 0.895). These results are suggestive of the involvement of Tuace gene duplication in resistance. Thus, monocrotophos resistance in TSSMs appears to have evolved through a combination of mutation accumulation and extensive gene duplication. PMID- 20002215 TI - A diversity of putative carboxylesterases are expressed in the antennae of the noctuid moth Spodoptera littoralis. AB - Recent studies have suggested that pheromone-degrading enzymes belonging to the carboxylesterase family could play a role in the dynamics of the olfactory response to acetate sex pheromones in insects. Bioinformatic analyses of a male antennal expressed sequence tag library allowed the identification of 19 putative esterase genes expressed in the antennae of the moth Spodoptera littoralis. Phylogenetic analysis revealed that these genes belong to different insect esterase clades, defined by their putative cellular localization and substrate preferences. Interestingly, two of the 19 genes appeared to be antennal specific, suggesting a specific role in olfactory processing. This high esterase diversity suggested that the antennae are the location for intense esterase-based metabolism, against potentially a large range of exogenous and endogenous molecules. PMID- 20002216 TI - The effect of diet on the expression of lipase genes in the midgut of the lightbrown apple moth (Epiphyas postvittana Walker; Tortricidae). AB - We have identified lipase-like genes from an Epiphyas postvittana larval midgut EST library. Of the 10 pancreatic lipase family genes, six appear to encode active lipases and four encode inactive lipases, based on the presence/absence of essential catalytic residues. The four gastric lipase family genes appear to encode active proteins. Phylogenetic analysis of 54 lepidopteran pancreatic lipase proteins resolved the clade into five groups of midgut origin and a sixth of non-midgut lipases. The inactive proteins formed two separate groups with highly conserved mutations. The lepidopteran midgut lipases formed a ninth subfamily of pancreatic lipases. Eighteen insect and human gastric lipases were analysed phylogenetically with only very weak support for any groupings. Gene expression was measured in the larval midgut following feeding on five artificial diets and on apple leaves. The artificial diets contained different levels of triacylglycerol, linoleic acid and cholesterol. Significant changes in gene expression (more than 100-fold for active pancreatic lipases) were observed. All the inactive lipases were also highly expressed. The gastric lipase genes were expressed at lower levels and suppressed in larvae feeding on leaves. Together, protein motif analysis and the gene expression data suggest that, in phytophagous lepidopteran larvae, the pancreatic lipases may function in vivo as galactolipases and phospholipases whereas the gastric lipases may function as triacylglycerol hydrolases. PMID- 20002217 TI - Urinary bisphenol A concentrations and ovarian response among women undergoing IVF. AB - Bisphenol A (BPA) is a synthetic chemical used in the manufacture of materials present in many common consumer products. In experimental animals, BPA caused oocyte aneuploidy and reduced production of oestradiol. In a prospective cohort study, we investigated the association between urinary BPA concentrations and ovarian response among women undergoing in vitro fertilization (IVF) at the Massachusetts General Hospital (MGH) Fertility Center. The geometric mean of two specific-gravity (SG) adjusted urinary BPA concentrations collected during each IVF cycle was used as the cycle-specific BPA exposure level. BPA concentrations were measured using online solid phase extraction coupled to isotope dilution high-performance liquid chromatography-tandem mass spectrometry. Peak serum oestradiol was measured using the Elecsys Estradiol II immunoassay kit. Multivariable mixed effect models and Poisson regression models adjusting for correlation between multiple IVF cycles in the same woman were used to evaluate the association between urinary BPA concentrations and ovarian response, adjusting for age, BMI and day 3 follicle stimulating hormone (FSH) levels, a clinical measure of ovarian reserve. Urinary BPA concentrations were measured in 84 women (mean age 35.6 years) undergoing 112 IVF cycles; 23 women (27%) contributed more than one IVF cycle. BPA concentrations ranged from <0.4 to 25.5 microg/L (geometric mean 2.52 +/- SD 3.2); 15% of urine samples had concentrations <0.4 microg/L. Peak serum oestradiol levels correlated with the total number of oocytes retrieved per cycle (r = 0.65, p < 0.001). For each log unit increase in SG-BPA, there was an average decrease of 12% (95% CI: 4, 23%; p = 0.007) in the number of oocytes retrieved and an average decrease of 213 pg/ml (95% CI: -407, -20; p = 0.03) in peak oestradiol. BPA was detected in the urine of the majority of women undergoing IVF, and was inversely associated with number of oocytes retrieved and peak oestradiol levels. PMID- 20002219 TI - Oestrogenic mycotoxin exposures and precocious pubertal development. AB - Since the 1970s, there has been a worldwide scientific discussion on the potential health consequences of human exposure to endocrine disrupters: many environmentally persistent compounds are oestrogen agonists and/or androgen antagonists. Thus, they can dysregulate the hypothalamic-pituitary-gonadal axis potentially affecting human puberty timing. Zearalenone (ZEA) is a non-steroidal mycotoxin produced by Fusarium species on several grains. Despite its low acute toxicity and carcinogenicity, ZEA exhibits oestrogenic and anabolic properties in several animal species. ZEA food contamination is caused either by direct contamination of grains, fruits and their based-products or by 'carry-over' of mycotoxins in animal tissues, milk and eggs after intake of contaminated feedstuff. In addition, zeranol (alpha-ZAL), a resorcyl lactone derived from ZEA, has been widely used in the USA as a growth promoter to improve fattening rates in cattle. From 1978 to 1984, a great epidemic of premature thelarche and precocious puberty occurred in Puerto Rico. To explain this condition, it was suggested that dairy and meat products could be contaminated with anabolic oestrogens such as ZEA or alpha-ZAL. Subsequently, worldwide other groups have also reported causative associations between oestrogenic mycotoxins and development of early thelarche and/or precocious puberty in exposed children. In addition to animal data, epidemiological studies strongly support the hypothesis that human pubertal development may be induced by foetal/early or prepubertal exposure to oestrogenic compounds. Indeed, ZEA and its metabolites are able to adopt molecular conformation, which sufficiently resembles 17beta-oestradiol to allow it to bind to oestrogen receptors (ERs) in target cells exerting oestrogenic (agonist) actions. In this view, oestrogenic mycotoxins are suspected as triggering factor for precocious pubertal development at least in prepubertal exposed girls. PMID- 20002214 TI - Gene expression associated with changes in cold tolerance levels of the Antarctic springtail, Cryptopygus antarcticus. AB - The ability of the Antarctic microarthropod Cryptopygus antarcticus (Collembola, Isotomidae) to survive low temperatures has been well studied at the physiological level, with recent investigations indicating the importance of the moulting process in conferring this ability. This study investigated gene expression in groups of C. antarcticus that have distinct differences in their ability to survive low temperatures. A microarray containing c. 5400 C. antarcticus expressed sequence tags was used to investigate gene expression differences between groups of animals with different supercooling points (SCP), and to low temperatures close to their SCP. By demonstrating the involvement of moult-related genes in the differential survival of two groups of C. antarcticus with distinct SCP profiles, the results of this investigation add support to the suggestion that moulting plays a role in conferring cold tolerance in C. antarcticus. PMID- 20002218 TI - Birth defects in the sons and daughters of women who were exposed in utero to diethylstilbestrol (DES). AB - Prenatal exposure to diethylstilbestrol (DES) is associated with adverse health outcomes, including anatomic anomalies of the reproductive tract in women and of the genitourinary tract in men. The mouse model, which replicates many DES related effects seen in humans, suggests that prenatal DES exposure causes alterations that may affect the next generation of offspring. We asked women participating in a large, multi-centre study of prenatal DES exposure to report birth defects occurring among 4029 sons and 3808 daughters (i.e., the third generation). A subcohort of 793 third generation daughters was also queried for birth defects. We used logistic regression models to generate odds ratio and 95% confidence intervals for the association between prenatal DES exposure in the mother and birth defects in the offspring. Based on the mothers' reports, overall birth defects were elevated in the sons (OR = 1.53; 95% CI = 1.04, 2.23) and in the daughters (OR = 2.35; 95% CI = 1.44, 3.82). Most estimates of association were imprecise, but daughters appeared to have an excess of heart conditions (OR = 4.56; 95% CI = 1.27, 16.34). Our data suggest a possible association between the mother's prenatal DES exposure and birth defects in their offspring, particularly in daughters. We cannot, however, rule-out the possible influence of reporting bias. In particular, the exposed daughters' elevated risk of cardiac defects may be as a result of the underreporting of these conditions by unexposed mothers. PMID- 20002220 TI - Androgen action in the masculinization programming window and development of male reproductive organs. AB - We have shown previously that deficient androgen action within a masculinization programming window (MPW; e15.5-e18.5 in rats) is important in the origin of male reproductive disorders and in programming male reproductive organ size, but that androgen action postnatally may be important to achieve this size. To further investigate importance of the MPW, we used two rat models, in which foetal androgen production or action was impaired during the MPW by exposing in utero to either di(n-butyl) phthalate (DBP) or to flutamide. Reduced anogenital distance (AGD) was used as a monitor of androgen production/action during the MPW. Offspring were evaluated in early puberty (Pnd25) to establish if reproductive organ size was altered. The testes, penis, ventral prostate (VP) and seminal vesicles (SV) were weighed and penis length measured. Both DBP and flutamide exposure in the MPW significantly reduced penis, VP and SV size along with AGD at Pnd25; AGD and organ size were highly correlated. In DBP-, but not flutamide-, exposed animals, testis weight was also reduced and correlated with AGD. Intratesticular testosterone was also measured in control and DBP-exposed males during (e17.5) or after (e21.5) the MPW and related to AGD at e21.5. To evaluate the importance of postnatal androgen action in reproductive organ growth, the effect of combinations of prenatal and postnatal maternal treatments on AGD and penis size at Pnd25 was evaluated. In prenatally DBP-exposed animals, further postnatal exposure to either DBP or flutamide significantly reduced AGD and penis size in comparison with prenatal DBP exposure alone. In comparison, rats exposed postnatally to testosterone propionate after prenatal vehicle-exposure showed considerable increase in these parameters vs. controls. In conclusion, we show that the size of all male reproductive organs is programmed by androgen exposure in the MPW, but that growth towards this size is dependent on androgen action postnatally. PMID- 20002222 TI - Developmental regulation of the pheromone biosynthesis activating neuropeptide receptor (PBAN-R): re-evaluating the role of juvenile hormone. AB - Sex pheromone production in Helicoverpa armigera is regulated by pheromone biosynthesis-activating neuropeptide (PBAN), which binds to a G-protein coupled receptor at the pheromone gland. We demonstrate the temporal differential expression levels of the PBAN receptor (PBAN-R) gene, reaching peak levels at a critical period of 5 h post-eclosion. Previous studies implied a possible regulatory role for juvenile hormone (JH). We herein demonstrate that PBAN-R expression levels increase normally when females are decapitated or head-ligated, removing the source of JH, before peak transcript levels are reached. Similarly, sex pheromone production can be induced by PBAN in such decapitated females. These results indicate that up-regulation, at this critical time, is not dependent on JH originating from the head. Conversely, JH injected in vivo at this critical period significantly inhibits PBAN-R transcript levels. PMID- 20002221 TI - Yellow fever mosquito sterol carrier protein-2 gene structure and transcriptional regulation. AB - AeSCP-2, a sterol carrier protein, is involved in sterol trafficking in mosquitoes. The activity of the AeSCP-2 gene is important for mosquito development. An earlier study demonstrated that the transcription of this gene was upregulated by 20-hydroxyecdysone (20E) in cultured gut tissues. To investigate 20E-regulated transcription of the AeSCP-2 gene we truncated the upstream flanking region of AeSCP-2 gene and linked it to a reporter gene. The mosquito Aag-2 cell line was transfected with these promoter/reporter constructs and treated with 20E at various concentrations. Expression vectors of different transcription factors such as HR3 and beta FTZ-F1 were also co-transfected with the AeSCP-2 promoter/reporter constructs. The observed results demonstrated that varied combinations of transcription factors produce different promoter activities of the AeSCP-2 gene. This observation leads us to the conclusion that the partnership of transcription factors is crucial in regulating the transcriptional activity of the AeSCP-2 gene. PMID- 20002223 TI - Drosophila G9a is implicated in germ cell development. AB - In Drosophila ovaries, germline stem cells (GSCs) divide asymmetrically in the germaria to produce daughter GSCs and cystoblasts. Single cystoblasts differentiate to form germline cysts with 16 germline cells, all of which are connected by the fusome, a vesiculated structure critical for oocyte specification. We here show that histone H3K9 methyltransferase dg9a is associated with spectrosome/fusome formation in the germarium; dG9a(13414) mutant ovaries have disorganized spectrosome/fusome in about half the germaria, with reduced levels of hu-li tai shao and alpha-SPECTRIN proteins. We found that the amount of germline cells within cysts was reduced and that oocyte determination often failed in egg chambers of the dG9a(13414) mutant ovaries. These results suggest that a mutation in dG9a gene gives rise to anomalous spectrosome/fusome structures, which in turn lead to faulty germ-cell development in Drosophila ovaries. PMID- 20002224 TI - Functional analysis of the cis-acting elements responsible for the induction of the Cyp6a8 and Cyp6g1 genes of Drosophila melanogaster by DDT, phenobarbital and caffeine. AB - Many Drosophila cytochrome P450 or Cyp genes are induced by caffeine and phenobarbital (PB). To understand the induction mechanism, we created Drosophila S2 cell lines stably transformed with different luciferase reporter plasmids carrying upstream DNAs of Cyp6a8 allele of the resistant 91-R strain, and the 1.1 kb upstream DNAs of Cyp6g1 of the 91-R and the susceptible 91-C strains. Following 24 h treatment with dichlorodiphenyltrichloroethane (DDT), caffeine or PB, luciferase activity of all cell lines was determined. Results showed that the 0.1-kb DNA of Cyp6a8 and the upstream DNAs of Cyp6g1 from both strains are not induced by these chemicals in S2 cells. However, the 0.2-, 0.5- and 0.8-kb DNAs of Cyp6a8 showed 13-24-, 4-5- and 2.2-2.7-fold induction with caffeine, PB and DDT, respectively. These DNAs also showed a 2-3-fold synergistic effect of caffeine and PB but not of caffeine and DDT. The results suggest that the cis regulatory elements for all three chemicals are located within the -11/-199 DNA of Cyp6a8. Furthermore, caffeine and PB inductions appear to be mediated via different cis-elements, whereas caffeine and DDT induction may involve common regulatory elements. These stably transformed cell lines should help understand the mechanism of resistance-associated Cyp gene overexpression in Drosophila. PMID- 20002225 TI - Periurethral injection of autologous adipose-derived stem cells for the treatment of stress urinary incontinence in patients undergoing radical prostatectomy: report of two initial cases. AB - OBJECTIVES: To report a novel cell therapy using autologous adipose tissue derived stem cells (ADSC) for stress urinary incontinence caused by urethral sphincteric deficiency and the outcomes in two initial cases undergoing periurethral injection of stem cells for the treatment of urinary incontinence after radical prostatectomy. METHODS: Two patients with moderate stress incontinence after radical prostatectomy were enrolled. After liposuction of 250 mL of adipose tissue from the abdomen, we isolated ADSC from this tissue by using the Celution system. Subsequently, the isolated ADSC and a mixture of stem cells and adipose tissue were transurethrally injected into the rhabdosphincter and submucosal space of the urethra, respectively. Short-term outcomes during a 12 week follow-up were assessed by a 24-h pad test, a validated patient questionnaire, urethral pressure profile, transrectal ultrasonography, and magnetic resonance imaging. RESULTS: Urinary incontinence progressively improved after 2 weeks of injection up to 12 weeks in terms of decreased leakage volume in a 24-h pad test, decreased frequency and amount of incontinence, and improved quality of life as per the questionnaire. In urethral pressure profile, both maximum urethral closing pressure and functional profile length increased. Ultrasonography and magnetic resonance imaging showed sustained presence of the injected adipose tissue. Enhanced ultrasonography showed a progressive increase in the blood flow to the injected area. No significant adverse events were observed peri- and postoperatively. CONCLUSION: This preliminary study showed that periurethral injection of the autologous ADSC is a safe and feasible treatment modality for stress urinary incontinence. PMID- 20002226 TI - Endothelial dysfunction and erectile dysfunction in the aging man. AB - Penile erection is a vascular event that requires an intact endothelium to occur. A dysfunctional endothelium is an early marker for the development of atherosclerotic changes and can also contribute to the occurrence of acute cardiovascular events. The pathogenesis of both endothelial and erectile dysfunction (ED) is intimately linked through decreased expression and activation of endothelial nitric oxide (NO) synthase, and the subsequent blunted physiological actions of NO naturally occurring with aging. It is now well understood that ED is a symptom of underlying disease rather than a disease itself; for this reason in the near future both general practitioners, internal medicine practitioners and many specialists will have to interplay with sexual medicine. Aging in the man is also associated with several changes in arterial structure and function, part of them related to the decline of circulating levels of steroids, that is, testosterone and estradiol. These changes may be responsible, in part, for the lack of efficacy of ED treatments. The recent discovery that chronic administration of phosphodiesterase type 5 inhibitors may improve erectile and endothelial responsiveness of men previously non-responsive to on-demand regimes, and the knowledge that testosterone is one of the main modulators of the expression of penile phosphodiesterase type 5 isoenzyme, opens a new scenario in the treatment of men with ED and co-morbidities. The aim of this review is to discuss the pathophysiology of endothelial dysfunction and its relationship with ED in the aging male, and to suggest possible strategies to improve arterial function with regard to sexual dysfunctions. PMID- 20002227 TI - Semicircular canals and agility: the influence of size and shape measures. AB - The semicircular canals of the inner ear sense angular accelerations and decelerations of the head and enable co-ordination of posture and body movement, as well as visual stability. Differences of agility and spatial sensitivity among species have been linked to interspecific differences in the relative size of the canals, particularly the radius of curvature (R) and the ratio of the canal plane area to streamline length (P/L). Here we investigate the scaling relationships of these two size variables and also out-of-plane torsion in the three semicircular canals (anterior, posterior and lateral), in order to assess which is more closely correlated with body size and locomotor agility. Measurements were computed from 3D landmarks taken from magnetic resonance images of a diverse sample of placental mammals encompassing 16 eutherian orders. Body masses were collected from the literature and an agility score was assigned to each species. The R and P/L of all three semicircular canals were found to have highly significant positive correlations with each other and no statistical difference was found between the slope of 2P/L against R and 1. This indicated that, contrary to initial hypotheses, there is little difference between 2P/L and R as measures of semicircular canal size. A measure of the in-plane circularity of the canal was obtained by dividing 2P/L by R and out-of-plane torsion was measured as angular deviation from a plane of best fit. It was predicted that deviations from in-plane and out-of-plane circularity would increase at small body size due to the constraints of fitting a proportionately larger canal into a smaller petrous bone. However, neither measurement was found to have a significant correlation with body mass, indicating that deviations from circularity (both in-plane and out-of-plane) are not sufficient to alter P/L to an extent that would impact the sensitivity of the canals. 2P/L and R were both shown to be significantly correlated with locomotor agility. The posterior canal was the least correlated with agility, suggesting that it may be generally less closely aligned to the direction of movement than the anterior canal. Of the three canals, the lateral canal was the most highly correlated with agility. In particular, it could be used to distinguish between species that move in a largely 2D environment and those that locomote in 3D space (aerial, arboreal and aquatic species). This complements previous work suggesting that the lateral canal primarily commands navigation, whereas the vertical canals control reflex adjustments. It was also found that 2P/L is substantially better correlated with agility than is R in the lateral canal. This result is intriguing given the above finding that there is no statistical difference between 2P/L and R, and requires further investigation. PMID- 20002228 TI - Anatomical models and wax Venuses: art masterpieces or scientific craft works? AB - The art of wax modelling has an ancient origin but rose to prominence in 14th century Italy with the cult of votive artefacts. With the advent of Neoclassicism this art, now deemed repulsive, continued to survive in a scientific environment, where it flourished in the study of normal and pathological anatomy, obstetrics, zoology and botany. The achievement of having originated the creation of anatomical models in coloured wax must be ascribed to a joint effort undertaken by the Sicilian wax modeller Gaetano Giulio Zumbo and the French surgeon Guillaume Desnoues in the late 17th century. Interest in anatomical wax models spread throughout Europe during the 18th century, first in Bologna with Ercole Lelli, Giovanni Manzolini and Anna Morandi, and then in Florence with Felice Fontana and Clemente Susini. In England, the art of anatomical ceroplastics was brought to London from Florence by the sculptor Joseph Towne. Throughout the centuries many anatomical artists preferred this material due to the remarkable mimetic likeness obtained, far surpassing any other material. Independent of the material used, whether wood, wax or clay, anatomical models were always considered merely craft works confined to hospitals or faculties of medicine and have survived to this day only because of their scientific interest. Italian and English waxes are stylistically different but the remarkable results obtained by Susini and Towne, and the fact that some contemporary artists are again representing anatomical wax bodies in their works, makes the border that formerly separated art and craft indistinguishable. PMID- 20002229 TI - 3-D visualization and animation technologies in anatomical imaging. AB - This paper explores a 3-D computer artist's approach to the creation of three dimensional computer-generated imagery (CGI) derived from clinical scan data. Interpretation of scientific imagery, such as magnetic resonance imaging (MRI), is restricted to the eye of the trained medical practitioner in a clinical or scientific context. In the research work described here, MRI data are visualized and interpreted by a 3-D computer artist using the tools of the digital animator to navigate image complexity and widen interaction. In this process, the artefact moves across disciplines; it is no longer tethered to its diagnostic origins. It becomes an object that has visual attributes such as light, texture and composition, and a visual aesthetic of its own. The introduction of these visual attributes provides a platform for improved accessibility by a lay audience. The paper argues that this more artisan approach to clinical data visualization has a potential real-world application as a communicative tool for clinicians and patients during consultation. PMID- 20002230 TI - Edge behaviour in a minute parasitic wasp. AB - 1. Changes in the density of an organism near a boundary elements (edge effects) are often thought to be generated by changes in movement behaviour, but in most cases the mechanism underlying these effects is unknown. 2. We quantified the movement behaviour of a minute parasitic wasp, Anagrus columbi, in relation to edges in its habitat. This wasp attacks eggs of the planthopper Prokelisia crocea, which inhabits a wet prairie ecosystem composed of patches of its host plant prairie cordgrass (Spartina pectinata) interspersed within a matrix of mudflat, smooth brome (Bromus inermis) and native grasses. Two edge types are common in this system, cordgrass-mudflat and cordgrass-brome. 3. We conducted mark-recapture experiments in which wasps were released at the cordgrass-matrix edge and 50 cm within cordgrass or matrix, for both edge types. The marked wasps were recaptured using a grid of sticky traps. We fitted an advection-diffusion model to these data, yielding estimates of the diffusion rate and advection coefficient for cordgrass and matrix, for each release position and edge type. 4. The spatial distribution of wasps was well-described by the advection-diffusion model. The pattern suggests that marked wasps strongly biased their movements towards the edge when released in matrix, and to a lesser extent when released in cordgrass, while edge releases showed little bias. The advection coefficients were similar for the two edge types, as were the diffusion rates for the three substrates (cordgrass, brome, mudflat). The diffusive and advective components of movement were of comparable magnitude for matrix and cordgrass releases, suggesting equal amounts of directed and random movement. 5. Our results suggest the wasps are attracted to cordgrass patches across short distances, and that bias in their movements may concentrate them at the patch edge. Their edge behaviour is qualitatively different from that of the host insect. 6. The methodology described here could be readily adapted to other systems, where direct observations of movement are difficult but mark-recapture studies are feasible. PMID- 20002231 TI - Phenotypic correlates of hybridisation between red and sika deer (genus Cervus). AB - 1. Hybridisation with an invasive species has the potential to alter the phenotype and hence the ecology of a native counterpart. 2. Here data from populations of native red deer Cervus elaphus and invasive sika deer Cervus nippon in Scotland is used to assess the extent to which hybridisation between them is causing phenotypic change. This is done by regression of phenotypic traits against genetic hybrid scores. 3. Hybridisation is causing increases in the body weight of sika-like deer and decreases in the body weight of red-like females. Hybridisation is causing increases in jaw length and increases in incisor arcade breadth in sika-like females. Hybridisation is also causing decreases in incisor arcade breadth in red-like females. 4. There is currently no evidence that hybridisation is causing changes in the kidney fat weight or pregnancy rates of either population. 5. Increased phenotypic similarity between the two species is likely to lead to further hybridisation. The ecological consequences of this are difficult to predict. PMID- 20002232 TI - Giant right atrial diverticulum in an adult. AB - We describe a 32-year-old man with chest pain and a giant right atrial diverticulum who underwent surgical resection. Examination of resected atrial tissue showed extreme wall thinning, central aneurysmal formation, and focal endocardial fibrosis consistent with idiopathic dilatation of the right atrium. It is unclear what the best treatment of right atrial diverticulum are, nor are the risks of thromboembolism, arrhythmia, and rupture of the diverticulum clearly defined, either for patients as a whole or for symptomatic or asymptomatic subgroups. However, to reduce the risk of sudden death we recommend surgical resection of large diverticula. PMID- 20002233 TI - Herpes zoster following saphenous venectomy for coronary bypass surgery. PMID- 20002234 TI - Migrated sternal wire into the right ventricle: case report in cardiothoracic surgery. AB - Migrating wires as a result of sternal nonunion present clinical challenges. Cardiac Computed Tomographic Angiography helps locate these wires precisely, enabling detailed surgical planning. Sternal wire migration is an infrequent complication following median sternotomy. It is usually encountered among patients with sternal dehiscence. Understanding the location and spatial relationships of structures to the wire can be challenging. (64 slice) with high spatial and temporal resolution affords the possibility of enhancing presurgical planning. PMID- 20002235 TI - Congenital coronary artery fistula between right coronary artery and the coronary sinus. PMID- 20002236 TI - A case of systemic anaplastic large cell lymphoma with 'Hodgkin-like' appearance and skin involvement mimicking lymphomatoid papulosis. AB - We report a unique case of the CD30+ anaplastic large cell lymphoma (ALCL). A 44 year-old Japanese male presented with lymphadenopathy followed by skin involvement. Initially, a swollen cervical lymph node was recognized in 1989 and relapsed in 1991, which was histologically diagnosed as Hodgkin disease of nodular sclerotic type. In 1996, he presented ulcerative cutaneous nodules and swollen lymph nodes in his left inguinal region, which was then diagnosed with CD30+ ALCL. Both the lymphadenopathy and the skin lesion had been completely remitted by combining chemotherapy followed by radiotherapy. Thereafter, he had relapsing and remitting episodes of multiple papules and nodules on his face, trunk and extremities for 10 years. Repeated histopathological examination revealed similar tumor cell proliferation in the papules/nodules of the skin. Essentially similar immunohistochemical features, including CD30 and granzyme B expression, but not anaplastic lymphoma kinase (ALK), strongly suggested that all these tumors were sequential expression of one disease continued for 19 years. This case was finally diagnosed as CD30+ ALCL with unique skin involvement mimicking lymphomatoid papulosis (LyP). PMID- 20002237 TI - Metastatic adenocarcinoma of breast within a benign melanocytic nevus in the context of cutaneous breast metastatic disease. AB - Metastatic breast cancer of skin may be seen in a number of forms. It is rare to find it in association with a benign melanocytic nevus. We describe a patient with metastatic adenocarcinoma of breast within a benign melanocytic nevus and delineate this occurrence within the context of reviewing cutaneous breast metastatic disease. PMID- 20002239 TI - Cutaneous mesenchymal stem cells: status of current knowledge, implications for dermatopathology. AB - Stem cell biology is currently making its impact on medicine, which will probably increase over the next decades. It not only influences our therapeutic thinking caused by the enormous plasticity of stem cells but also affects diagnostic and conceptual aspects of dermatopathology. Although our knowledge of the keratinocytic stem cells located within the follicular bulge has exploded exponentially since their discovery in 1990, the concept of cutaneous mesenchymal stem cells (MSCs) is new. Described initially in 2001 in mice, MSCs later were also found in the human dermis. The connective tissue sheath and the papilla of the hair follicle probably represent the anatomical niche for cutaneous MSCs. In line with the cancer stem cell hypothesis, mutations of these cells may be the underlying basis of mesenchymal skin neoplasms, such as dermatofibrosarcoma protuberans. Furthermore, research on cutaneous MSCs may impact our thinking on the interaction of the epithelial component of skin neoplasms with their surrounding stroma. We are only in the early stages to recognize the importance of the potential contributions of cutaneous MSC research to dermatopathology, but it is not inconceivable to assume that they could be tremendous, paralleling the early discovery of the follicular bulge as a stem cell niche. PMID- 20002240 TI - Prurigo nodularis: systematic analysis of 58 histological criteria in 136 patients. AB - BACKGROUND: To date, there has been no systematic investigation of the detailed histological features of prurigo nodularis (PN) in a large cohort of patients. METHODS: This retrospective study includes skin biopsies of 136 patients (63 males, 73 females; mean age: 58.38 years) with PN. RESULTS: Highly characteristic for PN is the presence of thick compact orthohyperkeratosis; the hairy palm sign (folliculosebaceous units in nonvolar skin in conjunction with a thick and compact cornified layer, like that of volar skin); irregular epidermal hyperplasia or pseudoepitheliomatous hyperplasia; focal parakeratosis; hypergranulosis; fibrosis of the papillary dermis with vertically arranged collagen fibers; increased number of fibroblasts and capillaries; a superficial, perivascular and/or interstitial inflammatory infiltrate of lymphocytes, macrophages and, to a lesser extent, eosinophils and neutrophils. For comparison, histological findings in 45 patients (18 males, 27 females; mean, 55.64 years) with lichen simplex (LS) were studied. PN and LS, both of them scratch-induced, had 50 of 58 (86.2%) histological features in common. CONCLUSIONS: PN revealed a characteristic histological pattern. Absence of pseudoepitheliomatous hyperplasia or nerve fiber thickening, however, does not rule out the histological diagnosis of PN. A correlation of clinical and histological findings is necessary to reliably distinguish between PN and LS. PMID- 20002241 TI - A new view on psoriasis research efforts. PMID- 20002242 TI - Efficacy of capsaicin in the treatment of acne in one case: new pathophysiological perspectives. PMID- 20002243 TI - Treatment of refractory adult-onset pityriasis rubra pilaris with TNF-alpha antagonists: a case series. AB - BACKGROUND: Pityriasis rubra pilaris (PRP) is a rare inflammatory dermatosis with frequent clinical presentation as erythroderma. Conventional systemic treatment is often unsatisfactory and limited by long-term toxicity. The use of tumour necrosis factor (TNF) antagonists has been reported previously in single cases, but lacking long-term follow-up or comparison between different biological agents. OBJECTIVES: To assess the long-term efficacy and safety of TNF-alpha antagonist, infliximab and etanercept, either in monotherapy or in combination therapy of severe, refractory adult-onset PRP. METHODS: Seven patients of adult onset PRP, six newly diagnosed type-I and 1 type-II, which were resistant or ineligible to conventional systemic treatment, received a single course of infliximab or etanercept therapy, alone or in combination with low-dose acitretin (>0.25 mg/kg/daily). After complete remission and treatment discontinuation, a follow-up period of 12 months was evaluated for relapses. RESULTS: Six patients obtained complete remission after a single course of anti-TNF-alpha therapy: mean therapy duration was 19.3 weeks (range 6-48 weeks). All patients obtained significant clearing (>75% of body surface area) of skin lesions at week 12. Two patients with marked keratoderma developed localized disease recurrence during treatment. During follow-up, only a single patient, affected by type II PRP, had disease relapse. CONCLUSIONS: Both TNF-alpha antagonists proved successful for the treatment of refractory, adult-onset PRP, yielding complete and persistent clinical responses in type-I PRP. Infliximab was associated with a more rapid onset of action, while treatment duration was comparable with etanercept. PRP type II warranted long-term therapy and showed relapse after drug discontinuation. PMID- 20002244 TI - Variable discrimination and asymmetric preferences in laboratory tests of reproductive isolation between cichlid colour morphs. AB - Mating behaviour affects reproductive isolation and phenotypic differentiation. In Lake Tanganyika, the cichlid fish Tropheus moorii diversified into numerous, currently allopatric colour variants. Allopatric isolation is periodically interrupted by dispersal and secondary contact during lake level fluctuations, making long-term differentiation partly dependent on assortative mating. Laboratory experiments with two moderately distinct morphs revealed assortative female preferences in one (Nakaku), but random mate choice in the other morph (Mbita). No discrimination was apparent between two subtly differentiated morphs (Chimba and Moliro). Tested against each other in a previous study, the highly distinct Moliro and Nakaku exhibited strong assortative preferences. The correlation between colour pattern similarity and mate discrimination suggests that allopatry and philopatric behaviour are less crucial for the maintenance of differentiation between highly distinct morphs than for more similar morphs. Interestingly, the asymmetric isolation in one pair of morphs is congruent with a pattern of unidirectional mitochondrial introgression between populations. PMID- 20002245 TI - Degeneration patterns of the olfactory receptor genes in sea snakes. AB - The sense of smell relies on the diversity of olfactory receptor (OR) repertoires in vertebrates. It has been hypothesized that different types of ORs are required in terrestrial and marine environments. Here we show that viviparous sea snakes, which do not rely on a terrestrial environment, have significantly lost ORs compared with their terrestrial relatives, supporting the hypothesis. On the other hand, oviparous sea snakes, which rely on a terrestrial environment for laying eggs, still maintain their ORs, reflecting the importance of the terrestrial environment for them. Furthermore, we found one Colubroidea snake (including sea snakes and their terrestrial relatives)-specific OR subfamily which had diverged widely during snake evolution after the blind snake Colubroidea snake split. Interestingly, no pseudogenes are included in this subfamily in sea snakes, and this subfamily seems to have been expanding rapidly even in an underwater environment. These findings suggest that the Colubroidea specific ORs detect nonvolatile odorants. PMID- 20002246 TI - Fitness measures in selection analyses: sensitivity to the overall number of offspring produced in a lifetime. AB - Age at first (alpha) and last (omega) breeding are important life-history traits; however, the direction and strength of selection detected on traits may vary depending on the fitness measure used. We provide the first estimates of lifetime breeding success (LBS) and lambda(ind) (the population growth rate of an individual) of European badgers Meles meles, by genotyping 915 individuals, sampled over 18 years, for 22 microsatellites. Males are slightly larger than females, and the opportunity for selection was slightly greater for males, as predicted. lambda(ind) and LBS both performed well in predicting the number of grand-offspring, and both detected selection for a late omega, until the age of eight. Differential selection (S'(alpha)) for an early alpha, however, was only detected using LBS, not with lambda(ind). In declining populations (lambda(ind) < 1) selection favours reproduction later in life, whereas early reproduction is selected in increasing populations (lambda(ind) > 1). As 41% of badgers were assigned only one offspring (lambda(ind) < 1), whereas 40% were assigned more than two (lambda(ind) > 1), this cancelled out S'(alpha) measured by lambda(ind). PMID- 20002247 TI - Fisher's fundamental theorem of inclusive fitness and the change in fitness due to natural selection when conspecifics interact. AB - Competition and cooperation is fundamental to evolution by natural selection, both in animals and plants. Here, I investigate the consequences of such interactions for response in fitness due to natural selection. I provide quantitative genetic expressions for heritable variance and response in fitness due to natural selection when conspecifics interact. Results show that interactions among conspecifics generate extra heritable variance in fitness, and that interacting with kin is the key to evolutionary success because it translates the extra heritable variance into response in fitness. This work also unifies Fisher's fundamental theorem of natural selection (FTNS) and Hamilton's inclusive fitness (IF). The FTNS implies that natural selection maximizes fitness, whereas Hamilton proposed maximization of IF. This work shows that the FTNS describes the increase in IF, rather than direct fitness, at a rate equal to the additive genetic variance in fitness. Thus, Hamilton's IF and Fisher's FTNS both describe the maximization of IF. PMID- 20002248 TI - Spatial regression techniques for inter-population data: studying the relationships between morphological and environmental variation. AB - Understanding the importance of environmental dimensions behind the morphological variation among populations has long been a central goal of evolutionary biology. The main objective of this study was to review the spatial regression techniques employed to test the association between morphological and environmental variables. In addition, we show empirically how spatial regression techniques can be used to test the association of cranial form variation among worldwide human populations with a set of ecological variables, taking into account the spatial autocorrelation in data. We suggest that spatial autocorrelation must be studied to explore the spatial structure underlying morphological variation and incorporated in regression models to provide more accurate statistical estimates of the relationships between morphological and ecological variables. Finally, we discuss the statistical properties of these techniques and the underlying reasons for using the spatial approach in population studies. PMID- 20002249 TI - Did the introduction of maize into Europe provide enemy-free space to Ostrinia nubilalis? Parasitism differences between two sibling species of the genus Ostrinia. AB - We examined whether maize offers enemy-free space (EFS) to its pest Ostrinia nubilalis, and may thereby have contributed to its divergence from the sibling species, Ostrinia scapulalis, feeding mainly on mugwort, when introduced into Europe five centuries ago. We collected Ostrinia larvae on maize (70 populations, 8425 individuals) and mugwort (10 populations, 1184 individuals) and recorded parasitism using both traditional (counting emerging parasitoids) and molecular methods (detection by specific polymerase chain reaction). The main parasitoid was Macrocentrus cingulum (Braconidae). On mugwort, parasitism was twice that on maize, and parasitoid-related mortality was 8 times higher. This suggests that maize affords substantial EFS to Ostrinia feeding on it. The lower Mortality:Infestation ratio in maize suggests that O. nubilalis' immune response might be stronger than that of O. scapulalis. If so, adapting to maize and diverging from O. scapulalis would decrease the impact of parasitism on O. nubilalis at both ecological and evolutionary levels. PMID- 20002250 TI - The evolution of South American endemic canids: a history of rapid diversification and morphological parallelism. AB - The origin of endemic South American canid fauna has been traditionally linked with the rise of the Isthmus of Panama, suggesting that diversification of the dog fauna on this continent occurred very rapidly. Nevertheless, despite its obvious biogeographic appeal, the tempo of Canid evolution in South America has never been studied thoroughly. This issue can be suitably tackled with the inference of a molecular timescale. In this study, using a relaxed molecular clock method, we estimated that the most recent common ancestor of South American canids lived around 4 Ma, whereas all other splits within the clade occurred after the rise of the Panamanian land bridge. We suggest that the early diversification of the ancestors of the two main lineages of South American canids may have occurred in North America, before the Great American Interchange. Moreover, a concatenated morphological and molecular analysis put some extinct canid species well within the South American radiation, and shows that the dental adaptations to hypercarnivory evolved only once in the South American clade. PMID- 20002251 TI - Avian colour perception predicts behavioural responses to experimental brood parasitism in chaffinches. AB - Hosts of cuckoos have evolved defences allowing them to discriminate and reject parasite eggs. Mechanisms of discrimination are mostly visually mediated, and have been studied using approaches that do not account for what the receiver (i.e. host) actually can discriminate. Here, for the first time we apply a perceptual model of colour discrimination to study behavioural responses to natural variation in parasite egg appearance in chaffinches Fringilla coelebs. Discrimination of parasite eggs gradually increased with increasing differences in chromatic contrasts as perceived by birds between parasite and host eggs. These results confirm that colour differences of the eggs as perceived by birds are important integral parts of a matching signal used by chaffinch hosts. PMID- 20002252 TI - Ecological niche modelling as an exploratory tool for identifying species limits: an example based on Mexican muroid rodents. AB - Niche conservatism theory suggests that recently diverged sister species share the same ecological niche. However, if the ecological niche evolves as part of the speciation process, the ecological pattern could be useful for recognizing cryptic species. In a broad sense systematists agree that the niche characters could be used for species differentiation. However, to date such characters have been ignored. We used the genetic algorithm for rule-set production for modelling the ecological niche as a means of inferring ecological divergence in allopatric populations of muroid rodents for which taxonomic identity is uncertain. Our results show that niche differentiation is significant in most of the identified phylogroups. The differentiation is likely associated with natural evolutionary units, which can be identified by applying species concepts based on phylogenetic and ecological patterns (e.g. phylogenetic, cohesive, evolutionary). Even so, the role of the niche partition within phylogenetic reconstruction may be a limited one. PMID- 20002253 TI - Antagonistic coevolution across productivity gradients: an experimental test of the effects of dispersal. AB - Coevolution commonly occurs in spatially heterogeneous environments, resulting in variable selection pressures acting on coevolving species. Dispersal across such environments is predicted to have a major impact on local coevolutionary dynamics. Here, we address how co-dispersal of coevolving populations of host and parasite across an environmental productivity gradient affected coevolution in experimental populations of bacteria and their parasitic viruses (phages). The rate of coevolution between bacteria and phages was greater in high-productivity environments. High-productivity immigrants ( approximately 2% of the recipient population) caused coevolutionary dynamics (rates of coevolution and degree of generalist evolution) in low-productivity environments to be largely indistinguishable from high-productivity environments, whereas immigration from low-productivity environments ( approximately 0.5% of the population) had no discernable impact. These results could not be explained by demography alone, but rather high-productivity immigrants had a selective advantage in low-productivity environments, but not vice versa. Coevolutionary interactions in high productivity environments are therefore likely to have a disproportionate impact on coevolution across the landscape as a whole. PMID- 20002254 TI - The group selection controversy. AB - Many thought Darwinian natural selection could not explain altruism. This error led Wynne-Edwards to explain sustainable exploitation in animals by selection against overexploiting groups. Williams riposted that selection among groups rarely overrides within-group selection. Hamilton showed that altruism can evolve through kin selection. How strongly does group selection influence evolution? Following Price, Hamilton showed how levels of selection interact: group selection prevails if Hamilton's rule applies. Several showed that group selection drove some major evolutionary transitions. Following Hamilton's lead, Queller extended Hamilton's rule, replacing genealogical relatedness by the regression on an actor's genotypic altruism of interacting neighbours' phenotypic altruism. Price's theorem shows the generality of Hamilton's rule. All instances of group selection can be viewed as increasing inclusive fitness of autosomal genomes. Nonetheless, to grasp fully how cooperation and altruism evolve, most biologists need more concrete concepts like kin selection, group selection and selection among individuals for their common good. PMID- 20002255 TI - Isolation by distance in a continuous population under stochastic demographic fluctuations. AB - The local density of individuals is seldom uniform in space and time within natural populations. Yet, formal approaches to the process of isolation by distance in continuous populations have encountered analytical difficulties in describing genetic structuring with demographic heterogeneities, usually disregarding local correlations in the movement and reproduction of genes. We formulate exact recursions for probabilities of identity in continuous populations, from which we deduce definitions of effective dispersal () and effective density (D(e)) that generalize results relating spatial genetic structure, dispersal and density in lattice models. The latter claim is checked in simulations where estimates of effective parameters obtained from demographic information are compared with estimates derived from spatial genetic patterns in a plant population evolving in a heterogeneous and dynamic habitat. The simulations further suggest that increasing spatio-temporal correlations in local density reduce and generally decrease the product , with dispersal kurtosis influencing their sensitivity to density fluctuations. As in the lattice model, the expected relationship between the product and the genetic structure statistic a(r) holds under fluctuating density, irrespective of dispersal kurtosis. The product D sigma(2) between observed census density and the observed dispersal rate over one generation will generally be an upwardly biased (up to 400% in simulations) estimator of in populations distributed in spatially aggregated habitats. PMID- 20002256 TI - Sexually antagonistic co-evolution: a model and an empirical test. AB - Models reveal that sexually antagonistic co-evolution exaggerates female resistance and male persistence traits. Here we adapt an established model by including directional sexual selection acting against persistence. We find similar equilibria to previous models showing that sexually antagonistic co evolution can be limited by counteracting sexual, as well as, natural selection. We tested the model using empirical data for the seaweed fly, Coelopa ursina, in which body size acts as a persistence and a resistance trait. Our model can generate continuous co-evolutionary cycles and stable equilibria, however, all simulations using empirically derived parameter estimates reach stable equilibria. Thus, stable equilibria might be more common in nature than continuous co-evolutionary cycles, suggesting that sexual conflict is unlikely to promote speciation. The model predicts male biased sexual size dimorphism for C. ursina, comparable with empirically observed values. Male persistence is shown to be more sensitive than female resistance to changes in model parameters. PMID- 20002257 TI - Radiographic recognition of dental implants as an aid to identifying the deceased. AB - This study was undertaken to determine if dental implants can be radiographically differentiated by company type to aid forensic identification of the deceased. Recognition of dental implants on intraoral radiographic images was assessed in a blind study using a radiographic examination guide to highlight differences between dental implants. Inter- and intra-examiner comparisons were conducted and a computer program (Implant Recognition System) was evaluated to see whether it improved the accuracy of implant recognition. The study found that dental implants could be radiographically differentiated by company type. The Implant Recognition System in its current form was of little benefit for radiographic assessment of dental implants for forensic odontologists. Prior knowledge of implant types, with a McNemar's statistical value of 92.9, proved to be most significant in identification. PMID- 20002258 TI - Accidental ligature strangulation by an ironing machine: an unusual case. AB - In this paper, we present a case of a 53-year-old woman who had her headscarf catch on the cylinder ironing machine in the laundry of the hospital where she worked. The hospital workers found the woman dead with her head stuck to the ironing machine. After the death scene investigation and autopsy were completed, it was determined that the death occurred as a result of accidental ligature strangulation. Accidental ligature strangulation in which an article of clothing is caught in such an electrical machine and strangles the wearer is very rare. This case highlights the fact that these kinds of machines can be hazardous to work around and that increased safety measures should be taken to insure worker safety; additionally, the people who use these machines should be educated on the potential hazards. PMID- 20002259 TI - Detection and classification of ignitable liquid residues using a fluorescence based vapor-sensitive microsphere array. AB - This paper describes the application of microsphere vapor sensing arrays to the detection of ignitable liquid (IL) vapors as both pure vapors and as residues (ILRs) on simulated fire debris samples. The temporal fluorescence response profile of the microsphere array generated a reproducible pattern unique to each analyte that could be used to classify subsequent sensor responses. This system, together with a support vector machine pattern recognition algorithm, was used to address several different IL and ILR classification scenarios. High classification accuracy (98%) was maintained over more than 200 vapor responses and the array was able to identify ILs when presented to the pattern classification algorithm within a dataset containing 11 other volatile compounds. Both burned and unburned IL treated samples were classified correctly greater than 97% of the time. These results indicate that microsphere vapor sensing arrays may be useful for the rapid identification of ILs and ILRs. PMID- 20002260 TI - Intercondylar eminences and their effect on the maximum length measure of the tibia. AB - Maximum length measurement of the tibia has been found to be variable both in description and implementation. Historically, the literature often excludes the intercondylar eminences from the tibia in metric analysis. This paper explores the quantitative effects of inclusion or exclusion of the eminences on the maximum length measure across ancestral population, age and sex in five human adult populations of American Whites and Blacks, two Native American samples, and East Asians. A Tukey's post hoc comparison was employed to determine the overall effect of inclusion the intercondylar eminences has on metric assessment of the tibia. Results show no significant effect on comparative analysis of the tibia by age or sex. However, the difference between sample means by ancestry is significant (p < 0.0001). These results pose interesting questions concerning the morphological differences between ancestral groups. This investigation prompts further study of population variation of the human knee. PMID- 20002261 TI - Signature profiling and classification of illicit heroin by GC-MS analysis of acidic and neutral manufacturing impurities. AB - The illicit manufacture of heroin results in the formation of trace level acidic and neutral impurities. These impurities are detectable in illicit heroin and provide valuable information about the manufacturing process used. The isolation, derivatization, and semiquantitative analysis of neutral and acidic heroin manufacturing impurities by programmed temperature vaporizing injector-gas chromatography-mass spectrometry (PTV-GC-MS) is described. Trace acidic and neutral heroin impurities were isolated from basic fractions using liquid-liquid extraction. Extracted impurities were treated with N-Methyl-N trimethylsilyltrifluoroacetamide followed by PTV-GC-MS analyses. Semiquantitative data were obtained using full scan mass spectrometry utilizing unique ions or ion combinations for 36 trace impurities found in crude and/or highly refined heroin samples. Minimum detection limits for acidic and neutral impurities were estimated to be at the 10(-7) level relative to total morphine. Over 500 authentic heroin samples from South America, Mexico, Southwest Asia, and Southeast Asia were analyzed. Classification of illicit heroin based on the presence or absence and relative amounts of acidic and neutral impurities is presented. PMID- 20002262 TI - A comparison of cleaning regimes for the effective removal of fingerprint deposits from brass. AB - Effective removal of fingerprint deposits is crucial for experimentation related to the corrosion of metals by fingerprint deposits. Such removal is also necessary prior to deposition of test fingerprints. The effectiveness of four regimes in removing fingerprint deposits from brass is considered. Sustained wiping of the deposit with a tissue at applied pressures of up to c. 1430 Pa or rubbing while the brass was immersed in acetone both failed to remove completely all traces of fingerprint deposits. Heating the brass to 600 degrees C was an effective remover; however, this also oxidized the surface of the metal except where inhibited by fingerprint deposits. The most effective regime, and the only one of the four that removed all traces of deposit without affecting the properties of the metal surface, was immersion in warm soapy water while rubbing with a tissue. We propose this as the preferred method for fingerprint removal. PMID- 20002263 TI - The examination of commercial printing defects to assess common origin, batch variation, and error rate. AB - The examination of printing defects, or imperfections, found on printed or copied documents has been recognized as a generally accepted approach for linking questioned documents to a common source. This research paper will highlight the results from two mutually exclusive studies. The first involved the examination and characterization of printing defects found in a controlled production run of 500,000 envelopes bearing text and images. It was concluded that printing defects are random occurrences and that morphological differences can be used to identify variations within the same production batch. The second part incorporated a blind study to assess the error rate of associating randomly selected envelopes from different retail locations to a known source. The examination was based on the comparison of printing defects in the security patterns found in some envelopes. The results demonstrated that it is possible to associate envelopes to a common origin with a 0% error rate. PMID- 20002264 TI - Usefulness of systematic histological examination in routine forensic autopsy. AB - The forensic community does not agree on the need to perform histological examination at forensic autopsy. The aim of our study was to determine the usefulness of systematic standard histology in forensic autopsies. A prospective study was carried out on 428 autopsy cases for which standard histological examination was systematic. Mechanism of death not shown by gross anatomic findings was discovered by histology in about 40% of the cases. Cause of death was established by only histology in 8.4% of the cases. Microscopic findings affected the manner of death in 13% of the cases. Histology provided complementary information about prior medical condition of the deceased in about 49% of the cases. Traumatic lesions were better documented by histology in about 22% of the cases. According to the results of our study, systematic standard histology for the main organs should be used in routine forensic autopsies. PMID- 20002265 TI - Homicide-suicide in Konya, Turkey between 2000 and 2007. AB - Homicide followed by the suicide of the murderer is a relatively rare lethal incident in which an individual kills another person and subsequently dies by suicide. Cases involving a homicide and a suicide in which death examinations and autopsies were performed at The Konya Branch of the Forensic Medicine Council between 2000 and 2007 were retrospectively investigated. During the studied time period, there were 10 homicide-suicide cases identified with 10 perpetrators killing 12 victims. Nine of the perpetrators were men and eight of the victims were women. The precipitating motive was an impending divorce in four of the cases. Two perpetrators were described as severely depressed, one had a diagnosis of antisocial personality disorder, and one was a pedophile who had reactive depression. Firearms were used in eight of the homicide cases and seven of the suicides. Constricting the use of firearms may reduce/prevent future homicide suicide cases, as it is the most commonly used method to carry out homicide suicides. PMID- 20002266 TI - The reliability of chronological age determination by means of mandibular third molar development in subjects in Croatia. AB - The aim of this study was to determine the development of the mandibular third molar and its relation to chronological age. The evaluated material consisted of 979 orthopantomograms of patients of Croatian Caucasian origin aged between 5.7 and 14.6 years. Third molar developmental stages were evaluated according to the stages proposed by Nolla. The frequencies of different stages of mineralization and the mean value of the mineralization of the mandibular third molars with regard to age, gender, and size of the mandible were determined and the coefficient of correlation determined between the age of the subject and the stage of development of the mandibular molars. Statistically significant correlation was determined between mineralization of the mandibular third molar and chronological age of the subjects (boys, mandibular left third molar r = 0.779, right third molar r = 0.793; girls, mandibular left third molar r = 0.746, right third molar r = 0.725). It can be concluded that the accuracy of age estimation based on Nolla's method is applicable for Croatian children. PMID- 20002267 TI - Palatine sutures as age indicator: a controlled study in the elderly. AB - Vault sutures have proven their low reliability for estimating age at death in individual forensic science cases. We broke down the palatine sutures of 134 skulls (with known sex and age at time of death) into 15 subparts and 5 stages of fusion to obtain a mean coefficient of obliteration (Cp) which was then linked to five age classes. We completed this study with multiple regression equations of total palatine suture scores. We compared our results with those obtained using the Mann method on the one hand and classically segmented and scored ectocranial suture age determination methods on the other. Palatine sutures generally do not estimate age at death any better than cranial vault sutures. Despite the partly subjective aspect of suture study, palatine suture observation contributes additional information to age-range estimation, especially in old and very old subjects where other methods lose their effectiveness. PMID- 20002268 TI - The differentiation of the volatile organic signatures of individuals through SPME-GC/MS of characteristic human scent compounds. AB - Human scent evidence is utilized as an investigative tool through canine scent discriminations based on the premise that human scent is an individualizing characteristic. This study describes the development of what is effectively a human scent barcode consisting of the relative ratios of an individual's "primary odor" compounds utilized to determine a reproducible and individualizing profile which can be stored in a searchable database for a proof of concept of human scent as a biometric measure. Triplicate hand odor samples were evaluated from 10 subjects utilizing solid phase micro-extraction gas chromatography/mass spectrometry (SPME-GC/MS) and compared via Spearman Rank Correlations. Narrowing the compounds considered for each subject to only those common in all three samples, or a subject's "primary odor constituents," produced a greater degree of both individualization and discrimination; at both correlation thresholds of 0.9 and 0.8, the individuals were correctly discriminated and identified in 99.54% of the cases. PMID- 20002269 TI - The response of skin to applied stress: investigation of bitemark distortion in a cadaver model. AB - Knowledge of distortional properties of skin is important in bitemark analysis. Thus, the response of skin to stress from bites was investigated. Four sets of models were created from the dentition of one individual. Anterior teeth were systematically removed to vary contact surface area. A biting apparatus was constructed with an integrated load cell. Forty-six bites were created perpendicular to Langer lines on six cadavers. Rate of force application and bite pressure were controlled. Metric/angular measurement and hollow volume overlays were employed. Distortion produced by each dentition was calculated and assessed. Results showed that as teeth impressed loose tissue, mesial/distal distance increased, angles of rotation flattened, and inter-canine distance lengthened. An opposite effect was seen in tight tissue. When the surface area of the dentition was reduced, a mixture of these effects was observed. Conclusions indicated that stiffness of the tissue was the most important variable in bitemark distortion. PMID- 20002270 TI - Effect of gas chromatography temperature program on the association and discrimination of diesel samples. AB - Five diesel samples were analyzed by gas chromatography-mass spectrometry (GC-MS) using six GC temperature programs, aiming to investigate the effect of temperature program on association and discrimination of the samples. Temperature programs varied by ramp rate and incorporated one- or two-step temperature ramps. Pearson product moment correlation coefficients and principal components analysis were used to evaluate differences in discrimination among the diesel samples afforded by each temperature program, based on the total ion chromatogram (TIC) and selected extracted ion profiles (EIPs). Association of diesel replicates and discrimination among samples based on the TIC and aromatic EIP were similar for all temperature programs based on scores plots. The alkane EIP was not useful in discriminating samples regardless of temperature program, because of similar alkane content of the diesel samples. The association and discrimination of diesel samples was largely unaffected by temperature programs that incorporated one- or two-step temperature ramps. PMID- 20002271 TI - Gamma irradiation as a biological decontaminant and its effect on common fingermark detection techniques and DNA profiling. AB - The use of disease-causing organisms and their toxins against the civilian population has defined bioterrorism and opened forensic science up to the challenges of processing contaminated evidence. This study sought to determine the use of gamma irradiation as an effective biological decontaminant and its effect on the recovery of latent fingermarks from both porous and nonporous items. Test items were contaminated with viable spores marked with latent prints and then decontaminated using a cobalt 60 gamma irradiator. Fingermark detection was the focus with standard methods including 1,2-indanedione, ninhydrin, diazafluoren-9-one, and physical developer used during this study. DNA recovery using 20% Chelex extraction and quantitative real-time polymerase chain reaction was also explored. Gamma irradiation proved effective as a bacterial decontaminant with D-values ranging from 458 to 500 Gy for nonporous items and 797-808 Gy for porous ones. The results demonstrated the successful recovery of latent marks and DNA establishing gamma irradiation as a viable decontamination option. PMID- 20002272 TI - A method for estimating sex using metric analysis of the scapula. AB - The most accurate and precise methods for the assessment of age and stature often require knowledge of sex. Thus, being able to correctly identify sex from skeletal remains is critical in the forensic context. The presence of the os coxae or skull can never be guaranteed, making the development of reliable methods of sex estimation using other skeletal elements necessary. Using a 724 individual calibration sample from the Hamann-Todd collection, this study identifies sexual dimorphism in the human scapula, and presents a new five variable discriminant function for sex estimation. The overall accuracy of this method proved to be 95.7% on the cross-validated calibration sample, 92.5% on an 80 individual test sample from the Hamann-Todd collection, and 84.4% on a 32 individual test sample from the skeletal collection of the Wichita State University Biological Anthropology Laboratory. Additionally, a slightly less accurate two-variable model was developed and has cross-validated accuracy of 91.3%. PMID- 20002273 TI - Ninhydrin thiohemiketals: basic research towards improved fingermark detection techniques employing nano-technology. AB - In the first part of a comprehensive research project towards more efficient application of nano-technology to fingerprint visualization, we investigated the possibility of more selective binding of gold nanoparticles (NP) to fingerprint material. We synthesized derivatives of ninhydrin and 1,2-indanedione containing loosely bound thiol groups. In particular: thiohemiketals (THK) of ninhydrin, and thioketals of 1,2-indanedione were prepared and tested as potential fingerprint reagents. By reacting ninhydrin with various thiols we were able to produce a series of novel THK, bearing the SR group always at C2. Ninhydrin THK reacted with amino acids to produce the expected Ruhemann's purple, and they also developed latent fingermarks on paper in a similar manner to ninhydrin. Ketals and thioketals derived from 1,2-indanedione reacted neither with amino acids nor with latent fingermarks. In the second part of the research, the thiols which are formed on the ridges as byproducts of the reaction with amino acids will be tested for their potential as stabilizers for gold NP that will become covalently bound to the fingerprint ridges. PMID- 20002274 TI - Medical findings and legal outcomes in sexually abused children. AB - A previous study published in 2000 on a small group of children concluded that the medical examination in cases of sexual child abuse seldom provided legal proof of sexual abuse [J Forensic Sci 45(2000):115-7]. The present consecutive study included children referred to the police for a forensic medical examination. A colposcope was used to evaluate the anogenital findings which were classified as normal, nonspecific, and abnormal. Four hundred and eighty-two children were included. Abnormal anogenital findings were found in 38% of the girls and 20% of the boys, but there was no relation between abnormal anogenital findings and the two legal outcomes: "appearing in court" and "being convicted." However, the age of the child turned out to be a more important factor in relation to legal outcome than the physical findings. The results of this study suggest that the child's statement and not the physical findings were important for legal outcome. PMID- 20002275 TI - Fingering a murderer: a successful anthropological and radiological collaboration. AB - We illustrate an interdisciplinary approach to identify a victim in a case with complex taphonomic and procedural issues. Burning, fragmentation, species commingling, and examination by multiple experts required anthropological preparation and analysis combined with radiographic adaptations to image and match trabecular patterns in unusually small, burned specimens. A missing person was last seen in the company of a reclusive female on a remote rural property. A warranted search found several burn sites containing human and animal bones. Fragment preparation, analysis, and development of a biological profile by anthropologists enabled examination by the odontologist, molecular biologist, and radiologist, and justified use of antemortem radiographs from one potential victim. Visual and radiological comparison resulted in a positive (later confirmed) identification of the victim by radiological matches of three carpal phalanges. Although some dimensional changes are expected with burning, morphological details were preserved, aided by selection of relatively intact, small bones for comparison. PMID- 20002276 TI - Multicolor real-time PCR genotyping of ABO system using displacing probes. AB - Rapid and informative ABO genotyping has become increasingly popular in forensic use. We developed a multiplex real-time polymerase chain reaction (PCR) approach to genotype ABO major groups and subgroups. Seven differently fluorophor-labeled displacing probes for O(1)(261delG), A(261G), A(796C/803C), B(796A/803C), O(2) (802G>A), A(2) (1059delC), and A(2) (1009A>G) were combined in one or two PCRs to determine either ABO major groups or subgroups. The method correctly detected 13 reference DNA samples. A blind test of 237 samples resulted in complete agreement with their phenotypes, and 110 of these 237 samples as well as with PCR-SSP method. The whole analysis could be finished in less than 100 min at substantially low material cost and the template DNA ranging from 0.16 to 500 ng per reaction could be quantitatively detected. Despite the limited informativeness of ABO genotyping, the developed methods could find application in rapid and inexpensive screening of forensic settings. PMID- 20002277 TI - Natural, unexpected deaths: reliability of a presumptive diagnosis. AB - We retrospectively analyzed 100 deaths because of suspicions and concerns expressed by the family. We compared the preautopsy cause of death, as determined by a thorough review of the clinical data and circumstances, to the autopsy derived cause of death. In the majority (91/100), the preautopsy and postautopsy proximate causes of death were in agreement. In 9%, the autopsy provided information that resulted in a proximate cause of death different than anticipated. In four instances, the manner of death also was incorrect and was determined to be an accident rather than the originally presumed natural. No homicide or suicide would have been misclassified. In another nine instances, where the premortem and postmortem proximate causes of death were in agreement, the autopsy provided a specific mechanism of death. With a quality initial medicolegal death investigation, a subset of sudden deaths in adults may be reliably certified without an autopsy. PMID- 20002278 TI - Editorial for JIDR special issue on Resilience and People with Intellectual Disabilities. PMID- 20002280 TI - Radiology reporting templates in oncology: a time for change. PMID- 20002281 TI - Reader practice in mammography screen reading in Australia. PMID- 20002282 TI - Assessment of referral patterns for CT KUB in a tertiary setting. AB - The purpose of this study was to assess the referral patterns and the use of unenhanced renal tract CT (CT KUB) for investigating patients presenting with clinically suspected renal colic. We retrospectively reviewed 500 consecutive CT KUB studies requested for suspected renal colic carried out at a single institution between December 2006 and July 2007. Follow-up radiology reports and discharge summaries on the hospital clinical Intranet were also reviewed. Studies were analysed for characteristics including patient demographics, referring clinical team, time of referral, final diagnosis and requirement for further imaging. The majority of requests were from Emergency (ED) or Urology Departments (49%, 245 out of 500, and 37%, 186 out of 500, respectively). The positive rate for urolithiasis was 67% (337 out of 500), the negative rate was 25% (123 out of 500), and 8% (40 out of 500) of patients had alternative significant findings. Female patients were more likely to have a negative study than male patients (35 versus 20%, P < 0.0001) and more likely to have alternative significant pathology (12 versus 6%, P < 0.0001). Patients referred by specialities other than Urology and ED were more likely to be female and have a negative or alternative finding (P < 0.0001). CT KUB is a widely used first method of investigation for patients with suspected renal colic with a high positive predictive value allowing rapid diagnosis and intervention. However, given the high rate of negative or alternative findings on CT KUB in young women, especially those referred by specialities other than ED or Urology, ultrasound should be considered as an alternative imaging method to minimise unnecessary radiation exposure. PMID- 20002283 TI - Gastrointestinal stromal tumours: a clinico-radiologic review from a single centre in South India. AB - Gastrointestinal stromal tumours (GISTs) are rare tumours but are the commonest mesenchymal neoplasms in the gastrointestinal tract. To our knowledge, there is no large case series in Asian countries in which a clinico-radiological descriptive analysis of these tumours has been carried out. In this retrospective study, we analysed our experience of 70 patients with histopathologically proven GISTs, who were presurgically investigated by using CT, and describe the demography, anatomical distribution, imaging features and clinical course of the GIST. We found an unusually large predominance of males in our study, stomach and small bowel appeared to have been involved similarly and small bowel tumours had a higher rate of metastases. We also highlight some unusual CT features of these tumours that we encountered during the study, such as the presence of metastatic lymphadenopathy and satellite nodules, relapse in appendices epiploicae of the bowel, metachronous liposarcoma, adrenal and lung metastases, multiplicity of lesions and aneurysmal dilatation of the bowel. Two of our patients also had multiple neurofibromas, whose association with GIST has been seen in earlier reports. To the best of our knowledge, this article presents one of the largest series of articles on GISTs, to date, in Asian countries. We conclude with a differential diagnosis of GIST, with salient features distinguishing each entity. PMID- 20002284 TI - Reader practice in mammography screen reporting in Australia. AB - Reader variability is a problem in mammography image reporting and compromises the efficacy of screening programmes. The purpose of this exploratory study was to survey reader practice in reporting screening mammograms in Australia to identify aspects of practice that warrant further investigation. Mammography reporting practice and influences on concentration and attention were investigated by using an original questionnaire distributed to screen readers in Australia. A response rate of 71% (83 out of 117) was achieved. Demographic data indicated that the majority of readers were over 46 years of age (73%), have been reporting on screening mammograms for over 10 years (61%), take less than 1 min to report upon a screening mammogram examination (66%), report up to 200 examinations in a single session (83%) and take up to 2 h to report one session (61%). A majority report on more than 5000 examinations annually (66%); 93% of participants regard their search strategy as systematic, 87% agreed that their concentration can vary throughout a session, 64% agreed that the relatively low number of positives can lead to lapses in concentration and attention and almost all (94%) participants agreed that methods to maximise concentration should be explored. Participants identified a range of influences on concentration within their working environment including volume of images reported in one session, image types and aspects of the physical environment. This study has provided important evidence of the need to investigate adverse influences on concentration during mammography screen reporting. PMID- 20002285 TI - Guidelines for patient selection and performance of carotid artery stenting. AB - The endovascular treatment of carotid atherosclerosis with carotid artery stenting (CAS) remains controversial. Carotid endarterectomy (CEA) remains the benchmark in terms of procedural mortality and morbidity. Consensus Australasian guidelines for the safe performance of CAS were developed using the modified Delphi consensus method of iterative consultation. Selection of patients suitable for CAS needs careful consideration of clinical and patho-anatomical criteria. Randomised controlled trials and pooled analyses have demonstrated that CAS is more hazardous than CEA. The CGSC therefore recommends that CAS should not be performed in the majority of patients requiring carotid revascularisation. There is currently no evidence to support CAS as a treatment for asymptomatic carotid stenosis. The use of distal protection devices during CAS remains controversial with increased risk of clinically silent stroke. The knowledge requirements for the safe performance of CAS include an understanding of the evidence base from randomised controlled trials, carotid and aortic arch anatomy and pathology, and recognition and management of periprocedural complications. It is critical that all patients being considered for a carotid intervention have adequate pre procedural neuroimaging and peri-procedural, independent, neurological assessment. Maintenance of proficiency in CAS requires active involvement in surgical/endovascular audit and continuing medical education programmes. These standards should apply in the public and private health-care settings. These guidelines represent the consensus of an intercollegiate committee in order to direct appropriate patient selection to perform CAS. Advances in endovascular technologies and the results of randomised controlled trials will guide future revisions of this document. PMID- 20002286 TI - Skeletal changes mimicking intrathoracic disease on chest radiographs. AB - Various chest radiographs are illustrated to demonstrate features where bony changes may mimic intrathoracic disease. To confirm the skeletal origin and nature, further conventional radiographs often suffice, and the need for CT or scintigraphy may therefore be obviated. At the time of presentation for radiography of the chest, further pertinent clinical details can be obtained from the patient by the department staff, as required. All previous radiographs and radiological reports should be readily available. In order to add to the educational value of the 13 cases illustrated, all are presented in quiz format, with the answers and further images included in the text. PMID- 20002287 TI - Locally invasive nodular lymphoid hyperplasia: radiological findings in a case of pulmonary and sinus disease. AB - Nodular lymphoid hyperplasia is a rare, benign, lymphoproliferative disease usually found in the gastrointestinal tract. It has never been reported in the sinuses. We present an unusual case of pulmonary nodular lymphoid hyperplasia with locally invasive sinus involvement and describe the CT, MRI and positron emission tomography findings. PMID- 20002288 TI - RE: differential diagnosis of T2 hyperintense spinal cord lesions. PMID- 20002289 TI - Pregnancy and radiotherapy: management options for minimising risk, case series and comprehensive literature review. AB - This article reviews the efficacy and safety of radiotherapy in patients with cancer who are pregnant. Our review provided extended follow-up results in nine cases, presents a technical discussion on measures taken to minimise foetal radiation exposure and provides a comprehensive summary of the literature. Nine patients who received radiotherapy while pregnant are described. The clinical presentation and outcomes of these and 100 additional cases identified on a systematic literature review are presented. Comparisons of scattered radiation doses from three linear accelerators are presented. The average maternal follow up in our series was 8.9 years with one patient having a recurrence of their astrocytoma. In terms of foetal outcome, there were one death in utero, one elective termination of pregnancy and one on which no data were available. Six children, on whom long-term follow-up (average 10.3 years) was obtainable, were in good health. Overall, there had been 109 cases of radiotherapy in pregnancy that met our search criteria with 13 adverse outcomes and a median follow-up of 37 months. Comparisons of three linear accelerators demonstrated significant differences in the amount of scattered radiation to the abdominal surface. In summary radiotherapy during pregnancy can be associated with a significant number of adverse outcomes. While it may be difficult for a patient not to attribute these effects to radiotherapy, it is also difficult to define the mechanisms by which radiotherapy would have caused them, if that were the case. PMID- 20002290 TI - Use of deodorants during adjuvant breast radiotherapy: a survey of compliance with standard advice, impact on patients and a literature review on safety. AB - Proscription of antiperspirant or deodorant use during adjuvant breast radiotherapy is common. The investigators were seeking an information base to facilitate design of an appropriate controlled trial of the use of deodorants during radiotherapy. The first component consisted of a survey of women after adjuvant breast radiotherapy seeking information about routine deodorant use and potential concern if deodorants were not permitted during radiotherapy. The second component comprised a literature search for any existing controlled evidence regarding harm from deodorant use during radiotherapy. Four hundred fourteen women completed surveys. Two hundred eighty recalled advice against deodorants. Two hundred ninety-nine women routinely used deodorants, 70% of whom used roll-on products. Forty-five continued deodorant use during radiation, 20 of these despite recalling advice not to wear a deodorant. Of the 233 women who routinely wore a deodorant but abstained during radiotherapy, 19% expressed a lot of concern about body odour and 45% were slightly concerned. Three controlled studies totalling 310 patients report specific deodorants versus no deodorant use which did not show statistically significantly increased skin reactions, but had only a small subset with axillary irradiation. The proscription of deodorant use during radiotherapy is of unproven benefit and causes body odour concern to the majority of women who are usual deodorant users. The next most appropriate trial would compare use of the usual deodorant versus no deodorant, would encompass a significant number of women with radiotherapy to the axilla or application of deodorant to irradiated skin areas, and include endpoints other than skin reaction alone. PMID- 20002291 TI - Prospective development of an individualised predictive model for treatment coverage using offline cone beam computed tomography surrogate measures in post prostatectomy radiotherapy. AB - The aim of this study is to prospectively evaluate and model surrogate explanatory variables (SEVs) of target coverage and rectal dose pertaining to soft tissue anatomy visualised on cone beam computed tomography (CBCT) for incorporation into post-prostatectomy treatment coverage verification protocols. Twenty post-prostatectomy patients treated with conformal prostate bed radiotherapy (64-74 Gy) underwent CBCT daily at fractions 1 to 5, and then weekly. Treatment coverage was defined on each CBCT using 'PTV95', percentage of the CBCT PTV covered by original treatment fields, and 'RECTD50', dose delivered to 50% of CBCT rectal volume by original treatment fields. Three candidate SEVs for treatment coverage were defined for each scan: anterior rectal wall movement, change in bladder length and bladder base movement. Both anterior rectal wall movement and increase in bladder length predicted for the decreased PTV95 (P < 0.001 for each). Anterior movement of the anterior rectal wall predicted for increased RECTD50 (P < 0.001). Predictive models for the PTV95 and RECTD50 that accept the significant SEVs as inputs were developed. We developed simple CBCT acquired soft tissue anatomic surrogate measures that signal changes in target coverage and rectal dose during post-prostatectomy radiotherapy. Conventional bony anatomy patient position verification protocols were inadequate in accounting for soft tissue target and organ variation seen with CBCT. PMID- 20002292 TI - Coronary stenting with the sirolimus-eluting stent in clinical practice: final results from the prospective multicenter German Cypher Stent Registry. AB - AIMS: Drug-eluting coronary stents (DES) have gained widespread use for the treatment of coronary artery disease. However, because of safety concerns and frequent "off-label" use data from "real life," registries are necessary to monitor indications and outcome of DES in daily clinical practice. METHODS AND RESULTS: We evaluated data from the German Cypher Stent Registry. A total of 10,894 patients treated with at least one sirolimus-eluting stent (SES) at 152 hospitals were included. Follow-up at a median of 6.4 months was available in 10,006 patients (92%). Median age was 64.8 years and 75.5% were male. Per lesion a mean of 1.09 +/- 0.41 SES were implanted with a mean length of 21.1 +/- 11.5mm. During follow-up, death rate was 1.8% and the rates of myocardial infarction or stroke were 2.1% and 0.5%. Any target vessel revascularization (TVR) was performed in 8.0% of patients. Independent predictors for death, myocardial infarction, or stroke were: cardiogenic shock, acute coronary syndromes, reduced left ventricular function, renal insufficiency, diabetes mellitus, advanced age, three-vessel disease, degree of stenosis, and prior myocardial infarction. Predictors for a TVR were: two- or three-vessel disease, target vessel = coronary bypass, advanced age, stent diameter, ostial lesions, indication in-stent restenosis, renal failure, and target vessel = left anterior descended artery. CONCLUSIONS: These results demonstrate that SES use in clinical practice is safe and effective. The main predictors of clinical events during follow-up are clinical parameters whereas as predictors of TVR mainly are angiographic parameters. PMID- 20002293 TI - Recirculating delivery improves myocardial cell engraftment. AB - INTRODUCTION: Despite pharmacological advances for heart failure, morbidity and mortality remain unacceptably high. As a result, alternative approaches such as cell therapy have been suggested to hold potential promise. However, a major obstacle is the optimization of cell delivery to the heart. Therefore, we investigated the efficacy of a percutaneous recirculation system for the delivery of cells to the heart. METHODS: Ovine fibroblasts were delivered to the ovine heart (3 x 10(7) cells) using the V-Focus system, a "closed" recirculatory system that draws blood from the coronary sinus and returns it to the coronary artery via an oxygenator, or intracoronary (IC) infusion, followed by a 2-hour recovery period. Animals were euthanized and cardiac tissue collected to determine presence of cells. RESULTS: There was a significant difference (P < 0.05) in the number of cells delivered to the heart by the V-Focus compared to direct coronary infusion for left ventricular freewall (V-Focus 1.39 +/- 0.63/mm(2), IC 0.11 +/- 0.06/mm(2)), septum (V-Focus 3.18 +/- 0.88/mm(2), IC 0.38 +/- 0.19/mm(2)), and right ventricle (V-Focus 0.46 +/- 0.23/mm(2), IC 0.05 +/- 0.04/mm(2)). CONCLUSIONS: These results suggest that potential therapeutic cells are optimally delivered to the large animal heart using the V-Focus cardiac delivery system in an ovine heart. PMID- 20002294 TI - Ferroportin is a manganese-responsive protein that decreases manganese cytotoxicity and accumulation. AB - Although manganese (Mn) is an essential trace element for human development and growth, chronic exposure to excessive Mn levels can result in psychiatric and motor disturbances, referred to as manganism. However, there are no known mechanism(s) for efflux of excess Mn from mammalian cells. Here, we test the hypothesis that the cytoplasmic iron (Fe) exporter ferroportin (Fpn) may also function as a Mn exporter to attenuate Mn toxicity. Using an inducible human embryonic kidney (HEK293T) cell model, we examined the influence of Fpn expression on Mn-induced cytotoxicity and intracellular Mn concentrations. We found that induction of an Fpn-green fluorescent protein fusion protein in HEK293T cells was cytoprotective against several measures of Mn toxicity, including Mn-induced cell membrane leakage and Mn-induced reductions in glutamate uptake. Fpn-green fluorescent protein mediated cytoprotection correlated with decreased Mn accumulation following Mn exposure. Thus, Fpn expression reduces Mn toxicity concomitant with reduced Mn accumulation. To determine if mammalian cells may utilize Fpn in response to increased intracellular Mn concentrations and toxicity, we assessed endogenous Fpn levels in Mn-exposed HEK293T cells and in mouse brain in vivo. We find that 6 h of Mn exposure in HEK293T cells is associated with a significant increase in Fpn levels. Furthermore, mice exposed to Mn showed an increase in Fpn levels in both the cerebellum and cortex. Collectively, these results indicate that (i) Mn exposure promotes Fpn protein expression, (ii) Fpn expression reduces net Mn accumulation, and (iii) reduces cytotoxicity associated with exposure to this metal. PMID- 20002295 TI - Characterization of Ca2+ signaling pathways in mouse adrenal medullary chromaffin cells. AB - In the present study, we characterized the Ca2+ responses and secretions induced by various secretagogues in mouse chromaffin cells. Activation of the acetylcholine receptor (AChR) by carbachol induced a transient intracellular Ca2+ concentration ([Ca2+](i)) increase followed by two phases of [Ca2+](i) decay and a burst of exocytic events. The contribution of the subtypes of AChRs to carbachol-induced responses was examined. Based on the results obtained by stimulating the cells with the nicotinic receptor (nAChR) agonist, 1,1-dimethyl-4 phenylpiperazinium iodide, high K(+) and the effects of thapsigargin, it appears that activation of nAChRs induces an extracellular Ca2+ influx, which in turn activate Ca(2+)-induced Ca2+ release via the ryanodine receptors. Muscarine, a muscarinic receptor (mAChRs) agonist, was found to induce [Ca2+](i) oscillation and sustained catecholamine release, possibly by activation of both the receptor- and store-operated Ca2+ entry pathways. The RT-PCR results showed that mouse chromaffin cells are equipped with messages for multiple subtypes of AChRs, ryanodine receptors and all known components of the receptor- and store-operated Ca2+ entry. Furthermore, results obtained by directly monitoring endoplasmic reticulum (ER) and mitochondrial Ca2+ concentration and by disabling mitochondrial Ca2+ uptake suggest that the ER acts as a Ca2+ source, while the mitochondria acts as a Ca2+ sink. Our results show that both nAChRs and mAChRs contribute to the initial carbachol-induced [Ca2+](i) increase which is further enhanced by the Ca2+ released from the ER mediated by Ca(2+)-induced Ca2+ release and mAChR activation. This information on the Ca2+ signaling pathways should lay a good foundation for future studies using mouse chromaffin cells as a model system. PMID- 20002296 TI - Occult hepatitis B virus infection: a covert operation. AB - Detection of occult hepatitis B requires assays of the highest sensitivity and specificity with a lower limit of detection of less than 10 IU/mL for hepatitis B virus (HBV) DNA and <0.1 ng/mL for hepatitis B surface antigen (HBsAg). This covert condition is relatively common in patients with chronic hepatitis C virus (HCV) that seems to exert some influence on the replicative capacity and latency of HBV. Detection of virus-specific nucleic acid does not always translate into infectivity, and the occurrence of primer-generated HBV DNA that is of partial genomic length in immunocompetent individuals who have significant levels of hepatitis B surface antibody (anti-HBs) may not be biologically relevant. Acute flares of alanine aminotransferase (ALT) that occur during the early phase of therapy for HCV or ALT levels that remain elevated at the end of therapy in biochemical nonresponders should prompt an assessment for occult hepatitis B. Similarly, the plasma from patients with chronic hepatitis C that is hepatitis B core antibody (anti-HBc) positive (+/-anti-HBs at levels of <100 mIU/mL) should be examined for HBV DNA with the most sensitive assay available. If a liver biopsy is available, immunostaining for hepatitis B surface antigen (HBsAg) and hepatitis B core antigen (HBcAg) should be contemplated and a portion of the sample tested for HBV DNA. This is another reason for optimal collection of a specimen (e.g. two passes with a 16-guage needle under ultrasound guidance). Transmission of HBV to immunosuppressed orthotopic liver transplant recipients by donors with occult hepatitis B (OHB) will continue to occupy the interests of the transplant hepatologist. As patients with OHB may have detectable HBV DNA in serum, peripheral blood mononuclear cells (PBMC) and/or liver that can be reactivated following immunosuppression or intensive cytotoxic chemotherapy, the patient needs to be either monitored or treated depending on the pretreatment serological results such as an isolated anti-HBc reaction or a detectable HBV DNA. PMID- 20002297 TI - Comparative serum proteomic analysis of patients with acute-on-chronic liver failure: alpha-1-acid glycoprotein maybe a candidate marker for prognosis of hepatitis B virus infection. AB - The acute-on-chronic liver failure (AoCLF) caused by hepatitis B virus (HBV) infection remains to be a challenge in clinics with a high mortality rate in China, and it is important to identify biomarkers to foresee the prognosis of patients with HBV. The current study analysed serum proteome changes of acute-on chronic liver failure as a result of acute exacerbation of chronic hepatitis B infection. Serum samples were collected from normal subjects (NS, n = 8), patients with chronic hepatitis B (CHB, n = 12) and patients with AoCLF (n = 12). After removal of albumin/IgG and ultramembrane centrifugation, serum proteins were separated by two-dimensional gel electrophoresis. Differentially expressed spots were identified by matrix-associated laser desorption ionization time-of flight tandem mass spectrometry. Through the removal of albumin/IgG and ultramembrane centrifugation, the well-resolved and reproducible two-dimensional gel electrophoresis (2-DE) profiles were obtained. A total of 23 proteins were identified on 2-DE profiles by their differential expression between the three cohorts. Mass spectrometry analysis resulted in the identification of 12 proteins unambiguously. Western blot analysis confirmed the proteomics results that the alpha1-acid glycoprotein (alpha1-AGP) levels decrease significantly in plasma of patients with AoCLF, but somewhat decreased in patients with chronic HBV. Further alpha1-AGP levels in bulk serum samples were measured by immune turbidimetry including normal subjects group (n = 25), acute hepatitis group (n = 36), chronic hepatitis group (n = 52) and AoCLF group (n = 48), the level of alpha1-AGP in AoCLF groups sharply decrease than other groups. Our study shows that alpha1-AGP may be a potential plasma biomarker for AoCLF diagnosis because of acute exacerbation of chronic hepatitis B infection. PMID- 20002298 TI - Clinical and virological factors associated with hepatitis B virus reactivation in HBsAg-negative and anti-HBc antibodies-positive patients undergoing chemotherapy and/or autologous stem cell transplantation for cancer. AB - We studied clinical outcome and clinico-virological factors associated with hepatitis B virus reactivation (HBV-R) following cancer treatment in hepatitis B virus surface antigen (HBsAg)-negative/anti-hepatitis B core antibodies (anti HBcAb)-positive patients. Between 11/2003 and 12/2005, HBV-R occurred in 7/84 HBsAg-negative/anti-HBcAb-positive patients treated for haematological or solid cancer. Virological factors including HBV genotype, core promoter, precore, and HBsAg genotypic and amino acid (aa) patterns were studied. Patients presenting with reactivation were men, had an hepatitis B virus surface antibody (HBsAb) titre <100 IU/L and underwent >1 line of chemotherapy (CT) significantly more frequently than controls. All were treated for haematological cancer, 3/7 received haematopoietic stem cell transplantation (HSCT), and 4/7 received rituximab. Using multivariate analysis, receiving >1 line of CT was an independent risk factor for HBV-R. Fatal outcome occurred in 3/7 patients (despite lamivudine therapy in two), whereas 2/4 survivors had an HBsAg seroconversion. HBV-R involved non-A HBV genotypes and core promoter and/or precore HBV mutants in all cases. Mutations known to impair HBsAg antigenicity were detected in HBV DNA from all seven patients. HBV DNA could be retrospectively detected in two patients prior cancer treatment and despite HBsAg negativity. HBV-R is a concern in HBsAg-negative/anti-HBcAb-positive patients undergoing cancer therapy, especially in males presenting with haematological cancer, a low anti-HBsAb titre and more than one chemotherapeutic agent. HBV DNA testing is mandatory to improve diagnosis and management of HBV-R in these patients. The role of specific therapies such as rituximab or HSCT as well as of HBV aa variability deserves further studies. PMID- 20002300 TI - High levels of circulating N-terminal pro-brain natriuretic peptide in patients with hepatitis C. AB - Many patients chronically infected by hepatitis C virus (HCV) experience symptoms like fatigue, dyspnea and reduced physical activity. However, in many patients, these symptoms are not proportional to the liver involvement and could resemble symptoms of chronic heart failure. To our knowledge, no study evaluated serum levels of N-terminal pro-brain natriuretic peptide (NT-proBNP) in a large series of patients with HCV chronic infection (HCV+). Serum NT-proBNP was assayed in 50 patients HCV+ and in 50 sex- and age-matched controls. HCV+ patients showed significantly higher mean NT-proBNP level than controls (P = 0.001). By defining high NT-proBNP level as a value higher than 125 pg/mL (the single cut-off point for patient under 75 years of age), 34% HCV+ and 6% controls had high NT-proBNP (Fisher exact test; P < 0.001). With a cut-off point of 300 pg/mL (used to rule out chronic heart failure in patients under 75 years of age) 10% HCV+ and 0 controls had high NT-proBNP (Fisher exact test; P = 0.056). With a cut-off point of 900 pg/mL (used for ruling in chronic heart failure in patients with age 50 75) 8% HCV+ patients and 0 controls had high NT-proBNP (Fisher exact test; P = 0.12). The study demonstrates high levels of circulating NT-proBNP in HCV+ patients compared to healthy controls. The increase of NT-proBNP may indicate the presence of a sub-clinical cardiac dysfunction. Further prospective studies quantifying these symptoms in correlation with echocardiography are needed to confirm this association. PMID- 20002299 TI - Role of Hepatitis C virus core protein in viral-induced mitochondrial dysfunction. AB - Hepatitis C virus (HCV) infection results in several changes in mitochondrial function including increased reactive oxygen species (ROS) production and greater sensitivity to oxidant, Ca(2+) and cytokine-induced cell death. Prior studies in protein over-expression systems have shown that this effect can be induced by the core protein, but other viral proteins and replication events may contribute as well. To evaluate the specific role of core protein in the context of viral replication and infection, we compared mitochondrial sensitivity in Huh7-derived HCV replicon bearing cells with or without core protein expression with that of cells infected with the JFH1 virus strain. JFH1 infection increased hydrogen peroxide production and sensitized cells to oxidant-induced loss of mitochondrial membrane potential and cell death. An identical phenomenon occurred in genome length replicons-bearing cells but not in cells bearing the subgenomic replicons lacking core protein. Both cell death and mitochondrial depolarization were Ca(2+) dependent and could be prevented by Ca(2+) chelation. The difference in the mitochondrial response of the two replicon systems could be demonstrated even in isolated mitochondria derived from the two cell lines with the 'genome-length' mitochondria displaying greater sensitivity to Ca(2+) -induced cytochrome c release. In vitro incubation of 'subgenomic' mitochondria with core protein increased oxidant sensitivity to a level similar to that of mitochondria derived from cells bearing genome-length replicons. These results indicate that increased mitochondrial ROS production and a reduced threshold for Ca(2+) and ROS-induced permeability transition is a characteristic of HCV infection. This phenomenon is a direct consequence of core protein interactions with mitochondria and is present whenever core is expressed, either in infection, full-length replicon bearing cells, or in over-expression systems. PMID- 20002301 TI - Chronic hepatitis B virus infection and rubella susceptibility in pregnant women. AB - Increased rubella susceptibility has been shown in subjects from the Asian Pacific region where chronic hepatitis B virus (HBV) infection is endemic. This study was performed to explore the relationship between chronic HBV infection and rubella susceptibility in the obstetric population. We conducted a retrospective cohort study on 50556 pregnant women delivered in a university obstetric unit from January 1998 to June 2008. The incidence of rubella susceptibility according to maternal HBV carrier status was examined. HBV infection and rubella susceptibility were found in 5105 (10.1%) and 6102 (12.1%) women, respectively. Rubella susceptibility was more common in women with HBV (13.1%vs 12.0%, P = 0.017), even after adjusting for other confounding factors (odds ratio 1.11, 95% confidence interval 1.01-1.21). Advancing age was associated with progressively decreasing odds of rubella susceptibility, from 0.48 at age 20-24 years to 0.34 at age >= 40 years in women without HBV infection, but had no effect in women with hepatitis B. In conclusion, our study is the first to demonstrate an association between chronic HBV infection with rubella susceptibility. Further studies are warranted to confirm whether chronic HBV infection, especially that acquired by vertical transmission, may impair the immune response to rubella vaccine or natural infection throughout the reproductive age. PMID- 20002302 TI - Functional pathway analysis of genes associated with response to treatment for chronic hepatitis C. AB - Chronic hepatitis C (CH-C) is among the most common causes of chronic liver disease. Approximately 50% of patients with CH-C treated with pegylated interferon-alpha and ribavirin (PEG-IFN-alpha + RBV) achieve a sustained virological response (SVR). Several factors such as genotype 1, African American (AA) race, obesity and the absence of an early virological response (EVR) are associated with low SVR. This study elucidates molecular pathways deregulated in patients with CH-C with negative predictors of response to antiviral therapy. Sixty-eight patients with CH-C who underwent a full course of treatment with PEG IFN-alpha + RBV were included in the study. Pretreatment blood samples were collected in PAXgeneTM RNA tubes. EVR, complete EVR (cEVR), and SVR rates were 76%, 57% and 41%, respectively. Total RNA was extracted from pretreatment peripheral blood mononuclear cells, quantified and used for one-step RT-PCR to profile 154 mRNAs. The expression of mRNAs was normalized with six 'housekeeping' genes. Differentially expressed genes were separated into up and downregulated gene lists according to the presence or absence of a risk factor and subjected to KEGG Pathway Painter which allows high-throughput visualization of the pathway specific changes in expression profiles. The genes were consolidated into the networks associated with known predictors of response. Before treatment, various genes associated with core components of the JAK/STAT pathway were activated in the cohorts least likely to achieve SVR. Genes related to focal adhesion and TGF beta pathways were activated in some patients with negative predictors of response. Pathway-centred analysis of gene expression profiles from treated patients with CH-C points to the Janus kinase-signal transducers and activators of transcription signalling cascade as the major pathogenetic component responsible for not achieving SVR. In addition, focal adhesion and TGF-beta pathways are associated with some predictors of response. PMID- 20002303 TI - Generation of cellular immune responses to HCV NS5 protein through in vivo activation of dendritic cells. AB - Chronic hepatitis C (HCV) infection is a substantial medical problem that leads to progressive liver disease, cirrhosis, and hepatocellular carcinoma (HCC). The aim of this study was to achieve sustained cellular immune responses in vivo to a HCV nonstructural protein using dendritic cell (DC)-based immunization approach. We targeted the HCV NS5 protein to DCs in vivo by injecting microparticles loaded with this antigen. The DC population was expanded in BALB/C mice (H-2(d) ) by hydrodynamic injection of a plasmid pUMVC3-hFLex expressing the secreted portion of the human Fms-like tyrosine kinase receptor-3 ligand (hFlt3). Mice were subsequently injected with microparticles coated with HCV NS5 protein via the tail vein. Cellular immune responses were determined with respect to secretion of INFgamma and IL2 by CD4(+) cells and cytotoxic T-lymphocyte (CTL) assays in vitro; inhibition of tumour cell growth was employed for the assessment of CD8(+) generated activity in vivo. We found that Flt3L treatment expanded the DC population in the spleen to 43%, and such cells displayed a striking upregulation of CD86 as well as CD80 and CD40 co-stimulating molecules. Viral antigen-specific T(H) 1 cytokine secretion by splenocytes was generated, and CTL activity against syngeneic NS5 expressing myeloma target cells was observed. In addition, these cells inhibited tumour growth indicating that NS5-specific robust CTL activity was operative in vivo. Thus, the capability of activating DCs in vivo using the methods described is valuable as a therapeutic vaccine strategy for chronic HCV infection. PMID- 20002304 TI - Serum hepcidin levels are related to the severity of liver histological lesions in chronic hepatitis C. AB - Hepcidin is synthesized in the liver and has a crucial role in iron homoeostasis. Its synthesis is up-regulated in chronic inflammation and iron excess. We examined the determinants of serum hepcidin and liver hepcidin mRNA levels and their association with histological lesions in patients with chronic hepatitis C (CHC) and healthy controls. We studied 96 patients with CHC and 30 controls. Serum hepcidin levels were measured by an in-house competitive ELISA. Hepcidin mRNA levels were determined by a one-step qRT-PCR in total RNA extracted from liver biopsy specimens of 27 patients with CHC and six disease controls. Histological lesions were evaluated according to Ishak's classification. Serum hepcidin was significantly lower in patients with CHC than healthy controls (14.6 +/- 7.3 vs 34.6 +/- 17.3 ng/mL, P < 0.001). In patients with CHC, serum hepcidin correlated positively with aspartate aminotransferase (r = 0.334, P = 0.001) and insulin resistance (r = 0.27, P = 0.016) and had a trend for correlation with alanine aminotransferase (r = 0.197, P = 0.057) and serum haemoglobin (r = 0.188, P = 0.067) but not with ferritin. A significant positive correlation was also found between serum hepcidin levels and both necroinflammation (r = 0.259, P = 0.011) and fibrosis (r = 0.214, P = 0.036). Serum hepcidin was among others an independent predictor of cirrhosis (odds ratio: 1.145, P = 0.039). Liver hepcidin mRNA levels did not differ between patients and controls and were relatively lower in patients with than without cirrhosis (19.3 +/- 21.7 vs 38.3 +/- 26.0, P = 0.067). Patients with CHC have reduced serum hepcidin levels, which correlate with worse necroinflammation and fibrosis. The previously mentioned observations suggest a viral effect on hepatic hepcidin production, but might also support its involvement in the inflammatory process. PMID- 20002305 TI - Risk factors for intrahepatic cholangiocarcinoma: a possible role of hepatitis B virus. AB - There are several established risk factors for intrahepatic cholangiocarcinoma (ICC), namely primary sclerosing cholangitis, fibropolycystic liver disease, parasitic infection, intrahepatic biliary stones and chemical carcinogen exposure. However, the majority of patients with ICC do not have any of these risk factors. Therefore, identification of other risk factors is warranted for the prevention and early detection of ICC. We evaluated the risk factors for ICC in a large-scale cohort study in the province of Osaka, Japan. This retrospective cohort study included 154,814 apparently healthy individual blood donors, aged 40 64 years at the time of blood donation in the period 1991-1993. The average observation period was 7.6 years, resulting in 1.25 million person-years of observation. Incident ICC cases were identified by linking the blood-donor database to the records in the population-based cancer registry for the province. There were 11 incident ICC cases during follow-up, with an incidence rate of 0.88 per 100,000 person-years. Compared with subjects aged 40-49 years, the subjects aged 50-54 years and 55-59 years had a significantly higher risk for ICC (hazard ratio [HR] = 5.90; 95%CI:1.08-32.31 and 11.07; 95%CI:1.98-61.79, respectively). Compared with those with ALT level of 19 Karmen Units (KU) or less, subjects with ALT level of 40 KU or higher had a significantly higher risk for ICC (HR: 8.30; 95%CI:1.47-46.83). Compared with those who tested negative for both HBsAg and anti-HCV, those who tested HBsAg-positive had a significantly higher risk for ICC (HR: 8.56; 95%CI: 1.33-55.20). Our results suggest that HBV infection and liver inflammation are independently associated with ICC development. These findings need to be verified by further large cohort studies. PMID- 20002306 TI - MxA expression as marker for assessing the therapeutic response in HCV genotype 4 Egyptian patients. AB - The prevalence of hepatitis C virus (HCV) infection varies across the world, with the highest number of infections reported in Egypt. Expression of the MxA gene has been found to be a reliable and sensitive marker for the induction of endogenous type I interferons (IFNs) during viral infections. This study examined the correlation of gene expression of MxA with the response to treatment with pegylated-IFN-alfa2b and ribavirin. Fifty patients with type 4 HCV and 20 healthy volunteers as controls were enrolled in a prospective study designed with strict inclusion criteria to nullify the effect of confounding variables and further minimize selection bias. Quantification of HCV-RNA and MxA gene by real-time PCR was performed for every patient, and quantification of MxA gene was performed for controls. There was a statistically significant difference between patients and control group as regards the quantity of MxA gene expression (P < 0.05) (Mann Whitney test) (P = 0.004). There was a statistically significant difference between responders and nonresponders (P < 0.05): responders showed a higher percentage of cases with initial MxA <2(6) (P < 0.05). We conclude that MxA protein expression is a sensitive biological marker for ongoing virus replication and presence of type 1 IFN. These results highlight the importance of the detection of MxA expression at the start of therapy as a factor for assessing the likelihood of HCV genotype 4 patients to achieving a sustained virological response to treatment with IFN-alpha2 in combination with ribavirin. PMID- 20002307 TI - Prevalence of chronic viral hepatitis in people of south Asian ethnicity living in England: the prevalence cannot necessarily be predicted from the prevalence in the country of origin. AB - The prevalence of hepatitis B and hepatitis C in immigrant communities is unknown. Immigrants from south Asia are common in England and elsewhere, and the burden of viral hepatitis in these communities is unknown. We aimed to determine the prevalence of viral hepatitis in immigrants from south Asia living in England, and we therefore undertook a community-based testing project in such people at five sites in England. A total of 4998 people attending community centres were screened for viral hepatitis using oral fluid testing. The overall prevalence of anti-hepatitis C virus (HCV) in people of south Asian origin was 1.6% but varied by country of birth being 0.4%, 0.2%, 0.6% and 2.7% in people of this ethnic group born in the UK, India, Bangladesh and Pakistan, respectively. The prevalence of hepatitis B surface antigen was 1.2%-0.2%, 0.1%, 1.5% and 1.8% in people of this ethnic group born in the UK, India, Bangladesh and Pakistan, respectively. Analysis of risk factors for HCV infection shows that people from the Pakistani Punjab and those who have immigrated recently are at increased risk of infection. Our study suggests that migrants from Pakistan are at highest risk of viral hepatitis, with those from India at low risk. As prevalence varies both by country and region of origin and over time, the prevalence in migrant communities living in western countries cannot be easily predicted from studies in the country of origin. PMID- 20002308 TI - Hepatic vein transit times of a microbubble agent in assessing response to antiviral treatment in patients with chronic hepatitis C. AB - Microbubble measurement of hepatic vein transit times (HVTT) may have the potential to assess severity of hepatitis C virus (HCV)-related liver disease, where there is a shorter HVTT with more severe disease. We investigated the utility of this test as a marker of response to antiviral treatment. Thirty-seven patients with biopsy-proven HCV-related disease undergoing antiviral treatment were studied. All had baseline scans and then repeat scans 6 months after the end of treatment. HVTT using Levovist were obtained from the right and middle hepatic veins, and the shorter time was used for analysis. The aspartate aminotransferase to platelet ratio index (APRI) scores were calculated retrospectively. There were seven patients with mild hepatitis, 23 with moderate/severe hepatitis and seven with cirrhosis. The mean baseline HVTT in responders +/- SE increased from 27.3 +/- 2.29 s to 33.5 +/- 2.8 s posttreatment (P = 0.01). In the 10 nonresponders, the HVTT remained the same; 43.3 +/- 9 s baseline compared to 44 +/- 7.8 s posttreatment (P = 0.84). This trend was also seen with the APRI score where in responders, the mean score decreased from 1.1 +/- 0.2 to 0.74 +/- 1 (P = 0.03) and in nonresponders, the score remained unchanged; 0.88 +/- 0.2 compared to 0.84 +/- 0.2 (P = 0.31). HVTT measurement lengthened, while APRI scores decreased in patients who responded to antiviral treatment while both remained the same, shortened (HVTT) or increased (APRI), respectively, in patients who were nonresponders. These results are encouraging and indicate that these tests could be potentially used as markers of response to treatment and could obviate the need for serial biopsies in antiviral future treatment studies. PMID- 20002309 TI - Silver enhances the in vitro antifungal activity of the saponin, CAY-1. AB - The fungicidal properties of purified CAY-1, dissolved silver ion and ethylenediamine tetraacetic acid (EDTA) separately were studied in vitro as were the abilities of silver and EDTA to enhance CAY-1 fungicidal properties. Non germinated and germinating conidia of Aspergillus flavus, Aspergillus fumigatus, Aspergillus niger, Fusarium verticillioides (Fusarium moniliforme), Fusarium oxysporum and Fusarium solani were incubated separately with CAY-1 (0-24.8 MUg ml(-1)), silver (0-111.1 MUg ml(-1)), and EDTA (0-2400 MUg ml(-1)). Controls consisted of non-germinated or germinated conidia in test medium. To assess combined activity, compounds, based on the sub-lethal doses of each as defined in the initial experiments, were combined and tested in bioassays. Controls for the mixed sets consisted of non-germinated or germinated conidia only or with the sub lethal CAY-1 test concentrations. The minimum inhibitory concentrations (MICs) for CAY-1 and silver, both separate and combined, were determined. Viability assays showed CAY-1 activity only against the germinating conidia of A. flavus, A. niger and F. solani. Silver was active against the germinating conidia of all fungi and the non-germinated conidia of F. oxysporum and F. solani. Combined silver and CAY-1 produced significant viability loss at concentrations not effective separately. EDTA was not fungicidal separately and did not enhance CAY 1 fungicidal properties. MIC data showed that CAY-1 plus silver had an additive effect. Results indicate that dissolved silver was fungicidal in vitro and enhanced the fungicidal properties of CAY-1 at concentrations ineffective when tested separately. PMID- 20002310 TI - Tinea incognito in children: 54 cases. AB - The incidence of tinea incognito (TI) appears to have increased over recent years, although no large series of cases has been reported in children. The aim of this study was to analyse the main epidemiological, clinical and microbiological characteristics of TI diagnosed in children in comparison with other tineas. We undertook a retrospective study of 818 tineas diagnosed in children in a referral hospital between 1977 and 2006, concentrating on TI. Of the 54 TI diagnosed, 85% were in the last 15 years. Most children were older than 9 years of age. The most usual clinical forms were tinea corporis (46.3%) and tinea faciei (38.9%). Topical steroids alone had been used to treat 68.5% of the cases. Direct examination was positive in 91.5% of the cases examined. Culture was positive in 85.2% of cases. The most frequently isolated dermatophyte was Trichophyton mentagrophytes (44.4%). This is the largest case series of childhood TI reported to date. TI has increased over recent years and important differences were found between these TI and the other tineas in children over the same period. PMID- 20002311 TI - Immunohistological intensity measurements as a tool to assess sarcolemma associated protein expression. AB - AIMS: The quantification of protein levels in muscle biopsies is of particular relevance in the diagnostic process of neuromuscular diseases, but is difficult to assess in cases of partial protein deficiency, particularly when information on protein localization is required. The combination of immunohistochemistry and Western blotting is often used in these cases, but is not always possible if the sample is scarce. We therefore sought to develop a method to quantify relative levels of sarcolemma-associated proteins using digitally captured images of immunolabelled sections of skeletal muscle. METHODS: To validate our relative quantification method, we labelled dystrophin and other sarcolemmal proteins in transverse sections of muscle biopsies taken from Duchenne muscular dystrophy and Becker muscular dystrophy patients, a manifesting carrier of Duchenne muscular dystrophy and normal controls. RESULTS: Using this method to quantify relative sarcolemmal protein abundance, we were able to accurately distinguish between the different patients on the basis of the relative amount of dystrophin present. CONCLUSIONS: This comparative method adds value to techniques that are already part of the diagnostic process and can be used with minimal variation of the standardized protocols, without using extra amounts of valuable biopsy samples. Comparative quantification of sarcolemmal proteins on immunostained muscle sections will be of use to establish both the abundance and localization of the protein. Moreover, it can be applied to assess the efficacy of experimental therapies where only partial restoration or upregulation of the protein may occur. PMID- 20002312 TI - Loss of polyubiquitin gene Ubb leads to metabolic and sleep abnormalities in mice. AB - AIMS: Ubiquitin performs essential roles in a myriad of signalling pathways required for cellular function and survival. Recently, we reported that disruption of the stress-inducible ubiquitin-encoding gene Ubb reduces ubiquitin content in the hypothalamus and leads to adult-onset obesity coupled with a loss of arcuate nucleus neurones and disrupted energy homeostasis in mice. Neuropeptides expressed in the hypothalamus control both metabolic and sleep behaviours. In order to demonstrate that the loss of Ubb results in broad hypothalamic abnormalities, we attempted to determine whether metabolic and sleep behaviours were altered in Ubb knockout mice. METHODS: Metabolic rate and energy expenditure were measured in a metabolic chamber, and sleep stage was monitored via electroencephalographic/electromyographic recording. The presence of neurodegeneration and increased reactive gliosis in the hypothalamus were also evaluated. RESULTS: We found that Ubb disruption leads to early-onset reduced activity and metabolic rate. Additionally, we have demonstrated that sleep behaviour is altered and sleep homeostasis is disrupted in Ubb knockout mice. These early metabolic and sleep abnormalities are accompanied by persistent reactive gliosis and the loss of arcuate nucleus neurones, but are independent of neurodegeneration in the lateral hypothalamus. CONCLUSIONS: Ubb knockout mice exhibit phenotypes consistent with hypothalamic dysfunction. Our data also indicate that Ubb is essential for the maintenance of the ubiquitin levels required for proper regulation of metabolic and sleep behaviours in mice. PMID- 20002313 TI - Discovery, evaluation and distribution of haplotypes of the wheat Ppd-D1 gene. AB - Ppd-D1 is one of the most potent genes affecting the photoperiod response of wheat (Triticum aestivum). Only two alleles, insensitive Ppd-D1a and sensitive Ppd-D1b, were known previously, and these did not adequately explain the broad adaptation of wheat to photoperiod variation. In this study, five diagnostic molecular markers were employed to identify Ppd-D1 haplotypes in 492 wheat varieties from diverse geographic locations and 55 accessions of Aegilops tauschii, the D genome donor species of wheat. Six Ppd-D1 haplotypes, designated I-VI, were identified. Types II, V and VI were considered to be more ancient and types I, III and IV were considered to be derived from type II. The transcript abundances of the Ppd-D1 haplotypes showed continuous variation, being highest for haplotype I, lowest for haplotype III, and correlating negatively with varietal differences in heading time. These haplotypes also significantly affected other agronomic traits. The distribution frequency of Ppd-D1 haplotypes showed partial correlations with both latitudes and altitudes of wheat cultivation regions. The evolution, expression and distribution of Ppd-D1 haplotypes were consistent evidentially with each other. What was regarded as a pair of alleles in the past can now be considered a series of alleles leading to continuous variation. PMID- 20002314 TI - Distribution and abundance of the introduced ectomycorrhizal fungus Amanita phalloides in North America. AB - Despite a growing awareness of the global reach of ectomycorrhizal (EM) fungal introductions, little is known about the fate of introduced EM fungi in novel ranges. Using herbarium specimens, species distribution models, and field collections of sporocarps, root tips and extramatrical mycelia, we assessed the distribution and abundance of the European species Amanita phalloides in North America. There are two distinct ranges of the fungus, one along the West Coast (California to British Columbia) and the second on the East Coast (Maryland to Maine). As predicted by a species distribution model, the West Coast range is larger. Amanita phalloides is more frequently found in native forests on the West Coast than on the East Coast. At Point Reyes Peninsula in California, A. phalloides dominates community sporocarp biomass, and is frequent as root tips. In individual soil cores at Point Reyes, root tips of A. phalloides make up 50% of total root tip biomass. Hyphae of A. phalloides are frequent, but make up only 2% of total hyphal biomass. The contrasting patterns of the distribution and abundance of A. phalloides on the East and West Coasts of North America may influence both its future spread and its impacts. PMID- 20002315 TI - Homoeologous recombination in allopolyploids: the polyploid ratchet. AB - Polyploidization and recombination are two important processes driving evolution through the building and reshaping of genomes. Allopolyploids arise from hybridization and chromosome doubling among distinct, yet related species. Polyploids may display novel variation relative to their progenitors, and the sources of this variation lie not only in the acquisition of extra gene dosages, but also in the genomic changes that occur after divergent genomes unite. Genomic changes (deletions, duplications, and translocations) have been detected in both recently formed natural polyploids and resynthesized polyploids. In resynthesized Brassica napus allopolyploids, there is evidence that many genetic changes are the consequence of homoeologous recombination. Homoeologous recombination can generate novel gene combinations and phenotypes, but may also destabilize the karyotype and lead to aberrant meiotic behavior and reduced fertility. Thus, natural selection plays a role in the establishment and maintenance of fertile natural allopolyploids that have stabilized chromosome inheritance and a few advantageous chromosomal rearrangements. We discuss the evidence for genome rearrangements that result from homoeologous recombination in resynthesized B. napus and how these observations may inform phenomena such as chromosome replacement, aneuploidy, non-reciprocal translocations and gene conversion seen in other polyploids. PMID- 20002316 TI - The mechanics behind plant development. AB - Morphogenesis in living organisms relies on the integration of both biochemical and mechanical signals. During the last decade, attention has been mainly focused on the role of biochemical signals in patterning and morphogenesis, leaving the contribution of mechanics largely unexplored. Fortunately, the development of new tools and approaches has made it possible to re-examine these processes. In plants, shape is defined by two local variables: growth rate and growth direction. At the level of the cell, these variables depend on both the cell wall and turgor pressure. Multidisciplinary approaches have been used to understand how these cellular processes are integrated in the growing tissues. These include quantitative live imaging to measure growth rate and direction in tissues with cellular resolution. In parallel, stress patterns have been artificially modified and their impact on strain and cell behavior been analysed. Importantly, computational models based on analogies with continuum mechanics systems have been useful in interpreting the results. In this review, we will discuss these issues focusing on the shoot apical meristem, a population of stem cells that is responsible for the initiation of the aerial organs of the plant. PMID- 20002317 TI - Macroecological patterns of genetic structure and diversity in the aquatic moss Platyhypnidium riparioides. AB - Genetic diversity and structure are described in the aquatic moss Platyhypnidium riparioides to assess its dispersal ability at a regional scale and to determine whether patterns of genetic differentiation correlate with environmental variation. Variation at six nuclear microsatellite loci from 50 populations in southern Belgium was investigated through Mantel tests, partial Mantel tests and spatial analysis of molecular variance. Overall patterns of genotypic variation showed strong differentiation among populations at a regional scale (F(ST) = 0.57). The high values of F(IS) observed within populations at both the ramet and genet levels, and the higher proportion of ramets with the same genotype than expected by chance, all point to a strongly clonal or selfing mating system. A genetic discontinuity was identified between northern and southern groups of populations. Within each group, F(ST) and geographical distances were significantly correlated. Partial Mantel tests suggest that genetic and ecological distances are significantly correlated in the southern group. The results point to strong dispersal limitation at the landscape scale and suggest that the southern and northern groups experienced different histories. Within the former, the correlation between genetic and ecological variation is suggestive of reproductive isolation among ecotypes. PMID- 20002318 TI - Ecophysiological traits in C3 and C4 grasses: a phylogenetically controlled screening experiment. AB - Experimental evidence demonstrates a higher efficiency of water and nitrogen use in C(4) compared with C(3) plants, which is hypothesized to drive differences in biomass allocation between C(3) and C(4) species. However, recent work shows that contrasts between C(3) and C(4) grasses may be misinterpreted without phylogenetic control. Here, we compared leaf physiology and growth in multiple lineages of C(3) and C(4) grasses sampled from a monophyletic clade, and asked the following question: which ecophysiological traits differ consistently between photosynthetic types, and which vary among lineages? C(4) species had lower stomatal conductance and water potential deficits, and higher water-use efficiency than C(3) species. Photosynthesis and nitrogen-use efficiency were also greater in C(4) species, varying markedly between clades. Contrary to previous studies, leaf nitrogen concentration was similar in C(4) and C(3) types. Canopy mass and area were greater, and root mass smaller, in the tribe Paniceae than in most other lineages. The size of this phylogenetic effect on biomass partitioning was greater in the C(4) NADP-me species than in species of other types. Our results show that the phylogenetic diversity underlying C(4) photosynthesis is critical to understanding its functional consequences. Phylogenetic bias is therefore a crucial factor to be considered when comparing the ecophysiology of C(3) and C(4) species. PMID- 20002319 TI - A novel pepper (Capsicum annuum) receptor-like kinase functions as a negative regulator of plant cell death via accumulation of superoxide anions. AB - Plant receptor-like kinases belong to a large gene family. The Capsicum annuum receptor-like kinase 1 (CaRLK1) gene encodes a transmembrane protein with a cytoplasmic kinase domain and an extracellular domain. The CaRLK1 extracellular domain (ECD)-green fluorescent protein (GFP) fusion protein was targeted to the plasma membrane, and the kinase domain of the CaRLK1 protein exhibited autophosphorylation activity. CaRLK1 transcripts were more strongly induced in treatment with Xag8ra than in treatment with Xag8-13. Furthermore, infection with incompatible Xanthomonas campestris pv. vesicatoria race 3 induced expression of CaRLK1 more strongly than in the compatible interaction. Cell death caused by both a disease-forming and an HR-inducing pathogen was delayed in the CaRLK1 transgenic plants. Ectopic expression of CaRLK1 also induced transcripts of the lesion stimulating disease (LSD) gene, a negative regulator of cell death. Respiratory burst oxidase homolog (RBOH) genes were up-regulated in the transgenic plants compared with the wild type, as the concentration of the superoxide anion was increased. In contrast, the concentration of H(2)O(2) did not differ between the transgenic and wild-type plants. These results support the theory that the suppression of plant cell death by CaRLK1 is associated with consistent production of the superoxide anion and induction of the RBOH genes and the LSD gene, but not with the concentration of H(2)O(2). Thus, CaRLK1 may be a receptor of an as yet unidentified pathogen molecular pattern and may function as a negative regulator of plant cell death. PMID- 20002320 TI - Evolutionary rate variation, genomic dominance and duplicate gene expression evolution during allotetraploid cotton speciation. AB - Here, we describe the evolution of gene expression among a diversified cohort of five allopolyploid species in the cotton genus (Gossypium). Using this phylogenetic framework and comparisons with expression changes accompanying F(1) hybridization, we provide a temporal perspective on expression diversification following a shared genome duplication. Global patterns of gene expression were studied by the hybridization of petal RNAs to a custom microarray. This platform measures total expression for c. 42 000 duplicated genes, and genome-specific expression for c. 1400 homoeologs (genes duplicated by polyploidy). We report homoeolog expression bias favoring the allopolyploid D genome over the A genome in all species (among five polyploid species, D biases ranging from c. 54 to 60%), in addition to conservation of biases among genes. Furthermore, we find surprising levels of transgressive up- and down-regulation in the allopolyploids, a diminution of the level of bias in genomic expression dominance but not in its magnitude, and high levels of rate variation among allotetraploid species. We illustrate how phylogenetic and temporal components of expression evolution may be partitioned and revealed following allopolyploidy. Overall patterns of expression evolution are similar among the Gossypium allotetraploids, notwithstanding a high level of interspecific rate variation, but differ strikingly from the direction of genomic expression dominance patterns in the synthetic F(1) hybrid. PMID- 20002321 TI - Impact of transposable elements on the organization and function of allopolyploid genomes. AB - Transposable elements (TEs) represent an important fraction of plant genomes and are likely to play a pivotal role in fuelling genome reorganization and functional changes following allopolyploidization. Various processes associated with allopolyploidy (i.e. genetic redundancy, bottlenecks during the formation of allopolyploids or genome shock following genome merging) may allow accumulation of TE insertions. Our objective in carrying out a survey of the literature and a comparative analysis across different allopolyploid systems is to shed light on the structural, epigenetic and functional modifications driven by TEs during allopolyploidization and subsequent diploidization. The available evidence indicates that TE proliferation in the short or the long term after allopolyploidization may be restricted to a few TEs, in specific polyploid systems. By contrast, data indicate major structural changes in the TE genome fraction immediately after allopolyploidization, mainly through losses of TE sequences as a result of recombination. Emerging evidence also suggests that TEs are targeted by substantial epigenetic changes, which may impact gene expression and genome stability. Furthermore, TEs may directly or indirectly support the evolution of new functionalities in allopolyploids during diploidization. All data stress allopolyploidization as a shock associated with drastic genome reorganization. Mechanisms controlling TEs during allopolyploidization as well as their impact on diploidization are discussed. PMID- 20002322 TI - CNSB004 (Leconotide) causes antihyperalgesia without side effects when given intravenously: a comparison with ziconotide in a rat model of diabetic neuropathic pain. AB - OBJECTIVE: Leconotide is an omega-conotoxin that blocks neuronal voltage sensitive calcium channels. This study compared the antihyperalgesic potencies of leconotide and ziconotide given intravenously alone and in combinations with a potassium channel modulator flupirtine, given intraperitoneally, in a rat model of diabetic neuropathic pain. DESIGN: Rats were given streptozotocin (150 mg/kg ip) to induce diabetic neuropathy and hyperalgesia. Experiments were performed on diabetic rats with >or=30% hyperalgesia to noxious heat. Rats were given each conopeptide alone and with flupirtine. Open field activity monitoring and non invasive blood pressure measurements were used to define the maximum doses and combinations that caused no side effects. Doses in a range up to maximum no side effect doses were tested for antihyperalgesic effects in rats with hyperalgesia. RESULTS: The maximum no side effect dose of leconotide (2 mg/kg intravenously) caused 51.7% reversal of hyperalgesia compared with 0.4% for the highest no side effect dose of ziconotide (0.02 mg/kg; P < 0.001, one-way anova). Leconotide caused dose-related antihyperalgesic effects that were potentiated by coadministration with flupirtine at doses that were ineffective when given alone. Leconotide (0.02 mg/kg) and flupirtine (5 mg/kg) caused 25.3 +/- 7.6 and -6 +/- 9.5% reversal of hyperalgesia, respectively when given alone but in combination they caused 84.1 +/- 7.2% reversal of hyperalgesia (P < 0.01; one-way anova). No such interaction occurred with ziconotide. CONCLUSION: Leconotide could have wider clinical applications than ziconotide. Unlike ziconotide, powerful antihyperalgesia without side effects can be achieved by intravenous administration of leconotide thus avoiding the need for an intrathecal injection. PMID- 20002323 TI - Age and sex trends in long-term opioid use in two large American health systems between 2000 and 2005. AB - OBJECTIVE: To estimate recent age- and sex-specific changes in long-term opioid prescription among patients with chronic pain in two large American Health Systems. DESIGN: Analysis of administrative pharmacy data to calculate changes in prevalence of long-term opioid prescription (90 days or more during a calendar year) from 2000 to 2005, within groups based on sex and age (18-44, 45-64, and 65 years and older). Separate analyses were conducted for patients with and without a diagnosis of a mood disorder or anxiety disorder. Changes in mean dose between 2000 and 2005 were estimated, as were changes in the rate of prescription for different opioid types (short-acting, long-acting, and non-Schedule 2). PATIENTS: Enrollees in HealthCore (N = 2,716,163 in 2000) and Arkansas Medicaid (N = 115,914 in 2000). RESULTS: Within each of the age and sex groups, less than 10% of patients with a chronic pain diagnosis in HealthCore, and less than 33% in Arkansas Medicaid, received long-term opioid prescriptions. All age, sex, and anxiety/depression groups showed similar and statistically significant increases in long-term opioid prescription between 2000 and 2005 (35-50% increase). Per patient daily doses did not increase. CONCLUSIONS: No one group showed especially large increases in long-term opioid prescriptions between 2000 and 2005. These results argue against a recent epidemic of opioid prescribing. These trends may result from increased attention to pain in clinical settings, policy or economic changes, or provider and patient openness to opioid therapy. The risks and benefits to patients of these changes are not yet established. PMID- 20002324 TI - Estimating attractiveness for abuse of a not-yet-marketed "abuse-deterrent" prescription opioid formulation. AB - OBJECTIVE: The present study builds on research to model abusers' perceptions of particular analgesics' attractiveness for abuse and extends these methods to derive an estimate of attractiveness for abuse of a not-yet-marketed abuse deterrent formulation (ADF) of a prescription opioid (Remoxy), Pain Therapeutics, Inc., San Mateo, CA, and King Pharmaceuticals, Inc., Bristol, TN). In a previous study, the Opioid Attractiveness Technology Scaling (OATS) method identified, from a drug abuser's point of view, the particular features of a prescription opioid relevant to its attractiveness for recreational use. A second online sample rated the extent to which these features applied to particular products they had actually used/abused. These data were used to model the abusers' overall preference for prescription opioids they had used/abused. DESIGN: In the present study, this method was applied to a not-yet-marketed ADF using substance abuse counselors as proxies for prescription opioid abusers. Thirty-eight counselors were given materials describing the new ADF along with four known products. RESULTS: Thirty-two counselors demonstrated sufficient agreement with abusers' ratings of the overall attractiveness of these drugs. The overall model yielded a significant pseudo R(2) of 0.15 (P < 0.001), with increasing model fit based on preferred route of administration, from swallowing whole (pseudo R(2) = 0.06; P < 0.001) and best for those who preferred to inject (pseudo R(2) = 0.40; P < 0.001). Data from a cross-validation group of 16 counselors/proxies were used to calculate the OATS scores for the five rated drugs and revealed significant differences between the ADF and OxyContin (Purdue Pharma LP, Stamford, CT), Percocet (Endo Pharmaceuticals, Chadds Ford, PA), and Vicodin (Abbott Laboratories, Abbott Park, IL), but not Talwin NX (Sanofi-aventis, Bridgewater, NJ), which was identified in the prior study as a highly unattractive drug for recreational purposes. CONCLUSIONS: The OATS method shows promise for providing pre-marketing estimates of attractiveness for abuse of not-yet-marketed ADFs. PMID- 20002325 TI - Measuring attractiveness for abuse of prescription opioids. AB - OBJECTIVE: Prescription opioids are the second most misused/abused drug in the United States behind only marijuana. Recreational prescription opioid users appear to prefer some products over others; however, the extent to which attributes of any particular formulation account for such preferences has yet to be determined. The Opioid Attractiveness Technology Scaling was developed to identify the particular features of a prescription opioid that are relevant to its attractiveness for recreational use, and to use these features to model attractiveness for recreational use of particular prescription opioid formulations. DESIGN: Four hundred and ninety-one self-reported recreational prescription opioid users identified 43 product features as being relevant to determining whether a product is "attractive" or "unattractive" for recreational use. Average ratings were used to determine appropriate weights to be applied to the features. A factor analysis yielded 10, highly differentiated factors. Five hundred and sixty-four prescription opioid abusers were then asked to rate the extent to which the 43 features identified in Study 1 were relevant to specific prescription opioid products they had used. RESULTS: Respondents provided an overall preference rating of these products and a model was created. A random intercept model yielded a significant pseudo R(2) of 0.14 (chi-square = 310.02, degrees of freedom [df] = 10, P < 0.001). The model fit least well, albeit significantly, for abusers who preferred to swallow the drug (pseudo R(2) = 0.06; chi-square = 55.52, df = 10, P < 0.001) and best for those who preferred to inject (pseudo R(2) = 0.37; chi-square = 199.34, df = 10, P < 0.001). CONCLUSIONS: The relevance of the model is discussed along with possible modifications that might allow prediction of "attractiveness" of "abuse deterrent" formulations that have not yet been marketed. PMID- 20002326 TI - Reactive oxygen species from type-I photosensitized reactions contribute to the light-induced wilting of dark-grown pea (Pisum sativum) epicotyls. AB - Type-II, singlet oxygen-mediated photosensitized damage has already been shown to occur in epicotyls of dark-germinated pea (Pisum sativum L.) seedlings upon illumination, resulting in fast turgor loss and wilting. In this study we show evidence that the palette of reactive oxygen species (ROS) is more complex. Hydrogen peroxide, superoxide and hydroxyl radicals are also formed, suggesting the occurrence of type-I reactions as well. Moreover, hydrogen peroxide injection into the epicotyls in the dark was able to provoke wilting directly. Formation of hydroxyl radicals could also be triggered by the addition of hydrogen peroxide in the dark, preferentially in the mid-sections where wilting occurs, showing that potential mediators of a Fenton reaction are present in the epicotyls, but unevenly distributed. Localization of light-inducible ROS formation fully (hydrogen peroxide) or partially (superoxide radicals) overlaps with the distribution of monomer protochlorophyllide complexes, showing that these pigment forms are capable of provoking both type-I and type-II reactions. PMID- 20002327 TI - Genotypic differences in thermotolerance are dependent upon prestress capacity for antioxidant protection of the photosynthetic apparatus in Gossypium hirsutum. AB - Numerous studies have illustrated the need for antioxidant enzymes in acquired photosynthetic thermotolerance, but information on their possible role in promoting innate thermotolerance is lacking. We investigated the hypothesis that genotypic differences in source leaf photosynthetic thermostability would be dependent upon prestress capacity for antioxidant protection of the photosynthetic apparatus in Gossypium hirsutum. To test this hypothesis, thermosensitive (cv. ST4554) and reportedly thermotolerant (cv. VH260) G. hirsutum plants were exposed to control (30/20 degrees C) or high-day temperature (38/20 degrees C) conditions during flowering and source leaf gas exchange, chlorophyll content and maximum photochemical efficiency (F(v)/F(m)) were measured for each treatment. The relationship between source leaf thermostability and prestress antioxidant capacity was quantified by monitoring the actual quantum yield response of photosystem II (PSII) (Phi(PSII)) to a range of temperatures for both cultivars grown under the control temperature regime and measuring antioxidant enzyme activity for those same leaves. VH260 was more thermotolerant than ST4554 as evidenced by photosynthesis and F(v)/F(m) being significantly lower under high temperature for ST4554 but not VH260. Under identical growth conditions, VH260 had significantly higher optimal and threshold temperatures for Phi(PSII) and glutathione reductase (GR; EC 1.8.1.7) activity than ST4554, and innate threshold temperature was dependent upon endogenous GR and superoxide dismutase (SOD; EC 1.15.1.1) activity. We conclude that maintaining a sufficient antioxidant enzyme pool prior to heat stress is an innate mechanism for coping with rapid leaf temperature increases that commonly occur under field conditions. PMID- 20002328 TI - Real-time monitoring of herbivore induced volatile emissions in the field. AB - When plants are damaged by herbivorous insects they emit a blend of volatile organic compounds (VOCs) which include a range or terpenoids and green leaf volatiles (GLVs) formed via different metabolic pathways. The precise timing of these emissions upon the onset of herbivore feeding has not been fully elucidated, and the information that is available has been mainly obtained through laboratory based studies. We investigated emissions of VOCs from Populus tremula L. xP. tremuloides Michx. during the first 20 h of feeding by Epirrita autumnata (autumnal moth) larvae in a field site. The study was conducted using Proton Transfer Reaction-Mass Spectrometry (PTR-MS) to measure emissions online, with samples collected for subsequent analysis by complementary gas chromatography-mass spectrometry for purposes of compound identification. GLV emission peaks occurred sporadically from the outset, indicating herbivore activity, while terpene emissions were induced within 16 h. We present data detailing the patterns of monoterpene (MT), GLV and sesquiterpene (SQT) emissions during the early stages of herbivore feeding showing diurnal MT and SQT emission that is correlated more with temperature than light. Peculiarities in the timing of SQT emissions prompted us to conduct a thorough characterization of the equipment used to collect VOCs and thus corroborate the accuracy of results. A laboratory based analysis of the throughput of known GLV, MT and SQT standards at different temperatures was made with PTR-MS. Enclosure temperatures of 12, 20 and 25 degrees C had little influence on the response time for dynamic measurements of a GLV or MT. However, there was a clear effect on SQT measurements. Elucidation of emission patterns in real-time is dependent upon the dynamics of cuvettes at different temperatures. PMID- 20002329 TI - Differential accumulation of volatile terpene and terpene synthase mRNAs during lavender (Lavandula angustifolia and L. x intermedia) inflorescence development. AB - Despite the commercial importance of Lavandula angustifolia Mill. and L. x intermedia Emeric ex Loisel floral essential oils (EOs), no information is currently available on potential changes in individual volatile organic compound (VOC) content during inflorescence development. Calyces were found to be the main sites of VOC accumulation. The 20 most abundant VOCs could be separated into three sub-groups according to their patterns of change in concentration The three groups of VOCs sequentially dominated the global scent bouquet of inflorescences, the transition between the first and second groups occurring around the opening of the first flower of the inflorescence and the one between the second and third groups at the start of seed set. Changes in calyx VOC accumulation were linked to the developmental stage of individual flowers. Leaves accumulated a smaller number of VOCs which were a subset of those seen in preflowering inflorescences. Their nature and content remained constant during the growing season. Quantitative real time polymerase chain reaction assessments of the expression of two terpene synthase (TPS) genes, LaLIMS and LaLINS, revealed similar trends between their patterns of expression and those of their VOC products. Molecular and chemical analyses suggest that changes in TPS expression occur during lavender inflorescence development and lead to changes in EO composition. Both molecular data and terpene analysis support the findings that changes in biosynthesis of terpene occurred during inflorescence development. PMID- 20002330 TI - Simultaneous determination of Rubisco carboxylase and oxygenase kinetic parameters in Triticum aestivum and Zea mays using membrane inlet mass spectrometry. AB - The lack of complete Rubisco kinetic data for numerous species is partly because of the time consuming nature of the multiple methods needed to assay all of the Rubisco parameters. We have developed a membrane inlet mass spectrometer method that simultaneously determines the rate of Rubisco carboxylation (v(c)) and oxygenation (v(o)), and the CO(2) and O(2) concentrations. Using the collected data, the Michaels-Menten equations for v(c) and v(o) in response to changing CO(2) and O(2) concentrations were simultaneously solved for the CO(2) (K(c)) and O(2) (K(o)) constants, the maximum turnover rates of the enzyme for CO(2) (kcat(CO2)) and O(2) (kcat(O2)) and the specificity for CO(2) relative to O(2) (S(c/o)). In the C(4) species Zea mays K(c) was higher but K(o) was lower compared with the C(3) species Triticum aestivum. The kcat(CO2) was higher and the kcat(O2) lower in Z. mays compared with T. aestivum and S(c/o) was similar in the two species. The V(omax)/V(cmax) was lower in Z. mays and thus did not correlate with changes in S(c/o). In conclusion, this mass spectrometer system provides a means of simultaneously determining the important Rubisco kinetic parameters, K(c), K(o), kcat(CO2,)kcat(O2) and S(c/o) from the same set of assays. PMID- 20002331 TI - Single-vessel flow measurements indicate scalariform perforation plates confer higher flow resistance than previously estimated. AB - During vessel evolution in angiosperms, scalariform perforation plates with many slit-like openings transformed into simple plates with a single circular opening. The transition is hypothesized to have resulted from selection for decreased hydraulic resistance. Previously, additional resistivity of scalariform plates was estimated to be small - generally 10% or less above lumen resistivity - based on numerical and physical models. Here, using the single-vessel technique, we directly measured the hydraulic resistance of individual xylem vessels. The resistivity of simple-plated lumens was not significantly different from the Hagen-Poiseuille (HP) prediction (+6 + or - 3.3% mean deviation). In the 13 scalariform-plated species measured, plate resistivity averaged 99 + or - 13.7% higher than HP lumen resistivity. Scalariform species also showed higher resistivity than simple species at the whole vessel (+340%) and sapwood (+580%) levels. The strongest predictor of scalariform plate resistance was vessel diameter (r(2) = 0.84), followed by plate angle (r(2) = 0.60). An equation based on laminar flow through periodic slits predicted single-vessel measurements reasonably well (r(2) = 0.79) and indicated that Baileyan trends in scalariform plate evolution maintain an approximate balance between lumen and plate resistances. In summary, we found scalariform plates of diverse morphology essentially double lumen flow resistance, impeding xylem flow much more than previously estimated. PMID- 20002332 TI - MtPM25 is an atypical hydrophobic late embryogenesis-abundant protein that dissociates cold and desiccation-aggregated proteins. AB - Late embryogenesis-abundant (LEA) proteins are one of the components involved in desiccation tolerance (DT) by maintaining cellular structures in the dry state. Among them, MtPM25, a member of the group 5 is specifically associated with DT in Medicago truncatula seeds. Its function is unknown and its classification as a LEA protein remains elusive. Here, evidence is provided that MtPM25 is a hydrophobic, intrinsically disordered protein that shares the characteristics of canonical LEA proteins. Screening protective activities by testing various substrates against freezing, heating and drying indicates that MtPM25 is unable to protect membranes but able to prevent aggregation of proteins during stress. Prevention of aggregation was also found for the water soluble proteome of desiccation-sensitive radicles. This inhibition was significantly higher than that of MtEM6, one of the most hydrophilic LEA protein associated with DT. Moreover, when added after the stress treatment, MtPM25 is able to rapidly dissolve aggregates in a non-specific manner. Sorption isotherms show that when it is unstructured, MtPM25 absorbs up to threefold more water than MtEM6. MtPM25 is likely to act as a protective molecule during drying and plays an additional role as a repair mechanism compared with other LEA proteins. PMID- 20002333 TI - Making the most of drought and salinity transcriptomics. AB - More than 100 different studies of plant transcriptomic responses to salinity or drought-related stress have now been published. Most of these use microarrays or related high-throughput profiling technologies. This compels us to ask three questions in review: (1) what has transcriptomics contributed to our understanding of stress physiology; (2) what limits the ability of transcriptomics to contribute to increases in stress tolerance; and (3) given these limits, what are the most appropriate uses of transcriptomics? We conclude that although microarrays are now a mature technology that accurately describes the transcriptome, the consistently low correlation between transcript abundance and other measures of gene expression imposes an inherent limitation that cannot be ignored. Further limitations on the relevance of transcriptomics arise in some cases from experimental practices related to the treatment regimen and the selection of tissue or germplasm. Nevertheless, there is good evidence to support the continued use of transcriptomics, especially emerging techniques such as RNA Seq, as a screening tool for candidate gene discovery. Microarrays can also be valuable in analysing the transcriptome per se (e.g. when describing the phenotype of a transcription factor mutant or discovering non-coding RNA species), and when integrated with other types of data including metabolomic analyses. PMID- 20002334 TI - Control of leaf growth by abscisic acid: hydraulic or non-hydraulic processes? AB - Abscisic acid (ABA) affects plant metabolism and water transfers via multiple mechanisms at cell, organ and whole plant levels. These mechanisms translate into contradictory effects on leaf growth, so the literature reports positive, null or negative effects of ABA on leaf growth upon water deficit. We review evidences based on genetic manipulations of ABA biosynthesis, feeding the plant with artificial ABA or partial root drying and provide elements to avoid confusions of effects. We propose that ABA has mainly three effects on growth. (i) Via its controlling effect on stomatal aperture and transpiration rate, an increased concentration of ABA tends to buffer the day-night alternations of leaf growth rate and the negative effect of evaporative demand. (ii) ABA tends to improve leaf growth via an increase in the conductance to water transfer in the plant as a result of increased tissue hydraulic conductivity. (iii) ABA has also a modest non-hydraulic effect which is negative in plants subjected to water deficit, either manipulated for ABA synthesis or fed with artificial ABA, but can be positive in well watered plants deficient of ABA. The overall effect of increasing ABA biosynthesis depends on the relative weight of each of these effects under different environmental scenarios. PMID- 20002335 TI - Smoking among women following heart transplantation: should we be concerned? AB - The serious detrimental effects of smoking after heart transplantation (HTX) are well established, but data that demonstrate the effects on female HTX recipients are scarce. The purpose of this study was to describe tobacco use, exposure to second hand smoke (ESHS), and health perceptions of female HTX recipients and examine relationships between these variables of interest and demographic and clinical characteristics of women following HTX. Seventy-two women (mean age, 54.3+/-12.7 years; mean time since transplant, 5.5+/-4.5 years) were enrolled from a single HTX center. Demographic and clinical data, tobacco use, ESHS, and health perceptions were obtained through self-report and chart reviews. Tobacco use was verified by measurement of urine cotinine levels. Twenty-four women were nonsmokers before and after HTX. Eighteen (37.5%) of the 48 women who were former smokers before HTX had returned to tobacco abuse. Only 4 of the 18 accurately reported their smoking behaviors. Forty percent of nonsmokers reported ESHS. Tobacco use and ESHS were highest among African American women. Forty percent of the sample perceived their health status as fair-poor; the remaining 60% reported good-excellent health. In a multivariate analysis, current tobacco use (odds ratio [OR], 5.20; confidence interval [CI], 3.83-9.13) and ESHS (OR, 1.82; CI, 1.17-2.82) were independent predictors of lower health perceptions. Although a majority of the female recipients who used tobacco ceased smoking before HTX, a substantial proportion demonstrated recurrent tobacco use after HTX. Our findings suggest the need for aggressive screening and risk factor interventions to promote smoking cessation before and after HTX in this unique population of female HTX recipients. PMID- 20002336 TI - The impact of B-type natriuretic peptide, New York Heart Association classification and depression on quality of life in nonhospitalized heart failure patients. AB - Healthcare providers should be concerned with improving the quality of life (QOL) of patients with heart failure (HF) because disease-specific QOL is linked to disease progression. The present study investigated the significance of elevated b-type natriuretic peptide (BNP), NYHA classification and depression to HF related QOL to develop better management strategies. Outpatient subjects with left ventricular systolic dysfunction (n=108; mean age=64.9+/-12) completed the self-administered Minnesota Living with Heart Failure questionnaire and the Center for Epidemiologic Studies Depression Scale. Functional status was measured using the New York Heart Association Classification (NYHA) and BNP concentrations were measured in plasma samples. Multiregression analysis determined that plasma BNP levels did not contribute significantly to the total QOL score while depression (r=0.63, t ratio=7.43, P<.0001) and NHYA class (r=0.47, t ratio=3.31, P<.001) were significant contributors. NYHA III subjects exhibited worse depression scores (II 15+/-7 and III: 22+/-10, P<.001) and elevated plasma BNP (II: 2.0+/-0.5 and III: 2.4+/-0.6, P<.001). Low-cost psychological assessments are recommended to evaluate depression and suggest that those HF patients with NYHA III be closely monitored for depression and reduced QOL. PMID- 20002337 TI - Psychosocial factors, quality of life, and psychological distress: ethnic differences in patients with heart failure. AB - Advances in treatment have prolonged life in heart failure (HF) patients, leading to increased attention to quality of life (QOL) and psychological functioning. It is not clear if ethnic differences exist in factors associated with psychological well-being. We examined psychosocial factors associated with depression and anxiety in 97 HF patients. Medical records were reviewed and patients (M age 53, 50% African American) completed surveys examining social support, coping, spirituality, and QOL for their association with depression and anxiety. Multiple regressions suggested that psychosocial factors were associated with psychological health. Patients with lower social support, lower meaning/peace and more negative coping reported greater depression; positive coping, and lower meaning/peace were associated with higher anxiety. Ethnicity stratified models suggested that spiritual well-being was associated with depression only among African Americans and QOL partially mediated this relationship. Findings suggest the importance of considering the unique psychosocial needs of diverse populations to appropriately target clinical interventions. PMID- 20002338 TI - Sexual concerns and sexual counseling in heart failure. AB - Patients with heart failure (HF) face significant challenges in maintaining quality of life (QOL), particularly for sexual intimacy. Although recommended for all cardiac patients, it has been suggested that few HF patients receive sexual counseling. This study explored sexual counseling needs, sexual concerns, and sexual activity using a descriptive survey with HF patients (n = 45), recruited from a HF clinic or cardiology office. Most (77%) had not discussed sexual concerns with a health care professional (HCP). Sexual concerns that were rated as occurring 'occasionally/frequently' included partner overprotectiveness (63%), partner fear of sex (36%), lack of sexual interest (42%), erectile problems (74%), orgasmic difficulties (51%). Frequency of sexual intercourse before HF to present was striking, with 53% reporting no sexual activity in the last 2 months compared with 11% before diagnosis of HF. HCPs must provide sexual counseling to HF patients and partners to enhance QOL and to assist in any adaptations to sexual activity. PMID- 20002339 TI - Visiting preferences of cardiovascular patients. AB - The purpose of this study is to compare differences in patient preferences for visitors during hospitalization for a cardiovascular event specifically comparing cardiac intensive care unit (CICU) patients to those on cardiac step down units (SDU). A convenience sample 63 subjects from the CICU, mean age of 61.2 +/- 14.4 years, 21 males and 41 females, were compared with 61 subjects, mean age of 67.9 +/- 14.4 years, 15 females and 46 males, on the "Patient's Perception of Visits in the Hospital" questionnaire. The CICU patient perceived higher value for visiting specifically because visitors assisted with interpretation of information (P<.02), were calming (P<.05), providing information about them to staff (P<.01), helped with care (P<.000), and reinforced treatments (P<.004). There were no differences in total stressor score between CICU and SDU patients, though the CICU patient worried more about their visitors traveling (P<.025). The SDU patient perceived that visitors were disruptive to rest (P<.001) and that visitors intensified their pain (P<.008). The majority of patients preferred unlimited visiting hours, with 3 visitors at a time, and some limits on children. Text data identified unlimited visiting for close family members, the benefit of support from visitors, and the stress of lengthy visits. These data provide voice to cardiovascular patient's perception of visitors. PMID- 20002340 TI - Massage therapy reduces tension, anxiety, and pain in patients awaiting invasive cardiovascular procedures. AB - OBJECTIVES: (1) To assess the efficacy of a 20 minute massage therapy session on pain, anxiety, and tension in patients before an invasive cardiovascular procedure. (2) To assess overall patient satisfaction with the massage therapy. (3) To evaluate the feasibility of integrating massage therapy into preprocedural practices. Experimental pretest-posttest design using random assignment. Medical cardiology progressive care units at a Midwestern Academic Medical Center. Patients (N=130) undergoing invasive cardiovascular procedures. The intervention group received 20 minutes of hands on massage at least 30 minutes before an invasive cardiovascular procedure. Control group patients received standard preprocedural care. Visual analogue scales were used to collect verbal numeric responses measuring pain, anxiety, and tension pre- and postprocedure. The differences between pre- and postprocedure scores were compared between the massage and standard therapy groups using the Mann-Whitney Wilcoxon's test. Scores for pain, anxiety, and tension scores were identified along with an increase in satisfaction for patients who received a 20-minute massage before procedure compared with those receiving standard care. This pilot study showed that massage can be incorporated into medical cardiovascular units' preprocedural practice and adds validity to prior massage studies. PMID- 20002341 TI - Coronary artery bypass graft surgery patient education: a systematic review. AB - Patient education post-coronary artery bypass graft (CABG) surgery is an essential component of nursing care aimed at assisting patients in caring for themselves at home, following discharge from hospital. While there has been a recent interest in conducting meta-analysis and systematic reviews on the effectiveness of patient education during the preoperative period, no review was found on the topic of postoperative CABG patient education. This systematic review addressed the clinically-relevant questions: what approach, mode, and dose is most effective in producing changes in CABG patient education? Studies were included in the systematic review if they met the following selection criteria: (1) the sample represented adult (> or =18 years) patients who underwent CABG surgery; (2) the educational intervention involved the provision of self-care information following surgery but before discharge from hospital; (3) the outcome assessed related to self-care behavior; and (4) the study report was published in English between 1986 and 2008. A descriptive synthesis was used to code and extract data on publication information, study design, sample size, and quality of study, as well as postoperative CABG teaching, self-care behavior performance, and demographic characteristics of the patients who participated in the studies. A quantitative synthesis consisted of a statistical approach, which was used to calculate the magnitude of the treatment effects on self-care behavior. Results indicated larger effect sizes for CABG patient education in which the content was individualized, and given in a combination of media on an individual basis, and in more than one session. PMID- 20002342 TI - Cardiac support devices. AB - Heart failure is an increasingly common condition in the United States and is associated with high mortality and burden to health care. It is a chronic condition that is characterized by progressive left ventricular enlargement. While medical therapy can slow the progression of left ventricular remodeling, surgical approaches to treatment have been developed to improve the survival and quality of life of heart failure patients. This article reviews the surgical procedures for left ventricular dysfunction and focuses on cardiac support devices as a new therapy for heart failure patients. The nursing care of patients with cardiac support devices will be presented and a case study will highlight practical points to help guide patient care. PMID- 20002343 TI - Lipoprotein-associated phospholipase A(2): a promising vascular-specific marker for screening cardiovascular risk? AB - Atherosclerosis is an inflammatory disease that can be assessed by circulating biomarkers. Lipoprotein-associated phospholipase A(2) (Lp-PLA(2)) is an enzyme produced in atherosclerotic plaque and is bound to low density lipoprotein and high-density lipoprotein cholesterol. It has a role in the pathogenesis of atherosclerosis by promoting vascular inflammation. It is emerging as a vascular specific marker and predictor of risk for cardiovascular disease (CVD) events. Increasing evidence from many prospective epidemiologic studies have shown that elevated levels of Lp-PLA(2) are associated with future CVD events. Measurement of Lp-PLA(2) in individuals may provide clinically relevant information about their future risk of CVD events. Pharmacologic therapies and/or risk factor modification could be initiated after identification of individuals at risk for CVD. This review provides an overview of the pathophysiology, epidemiologic evidence, and clinical utility of Lp-PLA(2). PMID- 20002344 TI - Nutrigenomics in cardiovascular disease: implications for the future. AB - Cardiovascular disease (CVD), the leading cause of morbidity and mortality worldwide, is a complex multifactorial disease which is influenced by environmental and genetic factors. There is substantial evidence on the relationship between diet and CVD risk. An understanding of how genetic variation interacts with the diet to influence CVD risk is a rapidly evolving area of research. Since diet is the mainstay of risk factor modification, it is important to consider potential genetic influences on CVD risk. Nutrigenomics is the study of the interaction between diet and an individual's genetic makeup. Single nucleotide polymorphisms are the key factors in human genetic variation and provide a molecular basis for phenotypic differences between individuals. Whole genome and candidate gene association studies are two main approaches used in cardiovascular genetics to identify disease-causing genes. Recent nutrigenomics studies show the influence of genotype on the responsiveness to dietary factors or nutrients that may reduce CVD risk. Nutrigenomics research is expected to provide the scientific evidence for genotype-based personalized nutrition to promote health and prevent chronic disease, including CVD. It is imperative that healthcare providers, including cardiovascular nurses, are trained in genetics to foster delivery of competent genetic- and genomic-focused care and to facilitate incorporation of this new knowledge into current clinical practice, education, and research. PMID- 20002345 TI - Hypertension in Thailand. PMID- 20002346 TI - Sepsis and cardiac disease: improving outcomes through recognition and management. PMID- 20002347 TI - What is the significance of T-wave inversion in the precordial leads? PMID- 20002352 TI - PKA-mediated Golgi remodeling during cAMP signal transmission. AB - Cyclic AMP (cAMP)-dependent protein kinase A (PKA) is part of the set of signaling proteins that are stably associated to the cytosolic surface of Golgi membranes in mammalian cells. In principle, Golgi-associated PKA could participate in either signal transduction events and/or the coordination of Golgi transport activities. Here, we show data indicating that although Golgi associated PKA is activated fast and efficiently during cell stimulation by an extracellular ligand it does not contribute significantly to cAMP signal transmission to the nucleus. Instead, most of the PKA catalytic subunits Calphaderived from the Golgi complex remain localized in the perinuclear cytoplasm where they induce changes in Golgi structural organization. Thus, in stimulated cells the Golgi complex appears collapsed, showing increased colocalization of previously segregated markers and exhibiting merging of different proximal cisternae within a single stack. In contrast, the trans-Golgi network remains as a separate compartment. Consequently, the rate of protein transport is increased whereas glycan processing is not severely affected. This remodeling process requires the presence of PKA activity associated to the Golgi membranes. Together these data indicate that Golgi-associated PKA activity is involved in the adaptation of Golgi dynamic organization to extracellular signaling events. PMID- 20002353 TI - Spiral coating of the endothelial caveolar membranes as revealed by electron tomography and template matching. AB - Caveolae are invaginations of the plasma membrane involved in multiple cellular processes, including transcytosis. In this paper we present an extensive 3-D electron tomographic study of the endothelial caveolar system in situ. Analysis of large cellular volumes of (high-pressure frozen, freeze-substituted and epon embedded) human umbilical vein endothelial cells (HUVECs) provided a notable view on the architecture of the caveolar system that comprises--as confirmed by 3-D immunolabeling for caveolin of 'intact' cells--bona fide caveolae, free plasmalemmal vesicles, racemose invaginations and free multi-caveolar bodies. Application of template matching to tomograms allowed the 3-D localization of caveolar membrane coatings in a robust manner. In this way we observed that bona fide endothelial caveolae, cryofixed and embedded in their cellular context, show a spiral organization of the coating as shown in the past for chemically fixed and freeze-etched caveolae from fibroblasts. Meticulous 3-D analysis further revealed that the coatings are distributed in triads of spirals over the caveolar bulb and neck. Remarkably, this coating distribution is consistently present over the membranes of the other members of the caveolar system in HUVECs. The novel observations that we present clarify the ultrastructural complexity of the 'intact' caveolar system, setting a detailed morphological basis for its functional diversity. PMID- 20002354 TI - Erythema nodosum in renal transplant recipients: multiple cases and review of literature. AB - We report 4 renal transplant recipients with erythema nodosum. Erythema nodosum is a cutaneous inflammatory reaction located on the anterior aspects of the lower extremities. It may be associated with a wide variety of diseases, including infections (as in Cases 1 and 2), sarcoidosis, rheumatologic diseases, inflammatory bowel diseases (as in Case 3), medications (as in Case 4), autoimmune disorders, pregnancy, and malignancies. Histopathologically, erythema nodosum is the stereotypical example of a mostly septal panniculitis with no vasculitis, and the inflammatory infiltrate in the septa varies with age of the lesion. In early lesions edema, hemorrhage, and neutrophils are responsible for the septal thickening, whereas fibrosis, peri-septal granulation tissue, lymphocytes, and multinucleated giant cells are the main findings in late stage. Etiological management - by anti-tuberculous therapy in Cases 1 and 2, by salazopyrin in Case 3, and by discontinuation of ciprofloxacin in Case 4 - was associated with regression. Erythema nodosum can develop in renal transplant patients who did not receive induction therapy, non-rejecters, and those with steroid-free protocols. Management of erythema nodosum should be directed to the underlying associated condition, which could be tuberculosis, inflammatory bowel disease, or drug related. PMID- 20002355 TI - Prevention of recurrent hepatitis B virus infection after liver transplantation: hepatitis B immunoglobulin, antiviral drugs, or both? Systematic review and meta analysis. AB - OBJECTIVES: To evaluate antiviral prophylaxis against hepatitis B virus (HBV) following liver transplantation. METHODS: Systematic review and meta-analysis. Clinical trials and comparative cohort studies comparing the use of hepatitis B immunoglobulin (HBIg), antivirals, or both following liver transplantation for HBV infection were included. The primary outcome was reappearance of hepatitis B surface antigen (HBsAg). Other outcomes included all-cause and HBV-related mortality, HB-related active liver disease, and reappearance of HBV DNA after transplantation. Relative risks (RR) with 95% confidence intervals (CIs) are reported. RESULTS: Twenty studies (22 comparisons) were included. Ten studies compared HBIg to combination treatment, 9 compared antivirals to combination treatment, and 3 compared lamivudine (LAM) to HBIg. Combination treatment reduced HBsAg reappearance (RR 0.28; 95% CI 0.12-0.66), and was superior to HBIg alone in all other outcome measures. Combination treatment was significantly better than antivirals in preventing reappearance of HBsAg (RR 0.31; 95% CI 0.22-0.44), even when low-dose HBIg was given. No significant difference was found between HBIg and LAM monotherapy for all measured outcomes. Major limitations with regard to comparability of the study groups in non-randomized trials were revealed. CONCLUSIONS: Combination treatment with HBIg and LAM reduced HBV recurrence following liver transplantation, compared with HBIg or LAM alone, and reduced mortality compared with HBIg alone. PMID- 20002356 TI - Pharmacokinetics of oral valganciclovir solution and intravenous ganciclovir in pediatric renal and liver transplant recipients. AB - In an open-label, prospective, pharmacokinetic assessment, we evaluated total drug exposure (area under the curve [AUC]) of intravenous (IV) ganciclovir (GCV) and oral (p.o.) valganciclovir when normalized for body surface area (BSA) in pediatric liver (n=20) and renal (n=26) transplant patients Reference doses for IV GCV (200 mg/m(2)) and p.o. valganciclovir (520 mg/m(2)) were based on adult doses, and adjusted for BSA initially, and BSA and renal function (estimated via creatinine clearance [CrCL]) thereafter. Renal transplant patients received GCV on days 1-2, valganciclovir 260 mg/m(2) on day 3, and valganciclovir 520 mg/m(2) on day 4. Liver transplant patients received twice daily GCV from enrollment to day 12, and then valganciclovir twice daily on days 13-14. GCV pharmacokinetics were described using a population pharmacokinetic approach. Type of solid organ transplant (kidney or liver) had no effect on GCV pharmacokinetics. Median GCV exposure following valganciclovir 520 mg/m(2) was similar to that with IV GCV, and to that reported in adults. Patients <5 years of age had AUC values approximately 50% of those compared with older age ranges; dosing based on both BSA and CrCL increased drug exposure in younger patients. A dosing algorithm based on BSA and CrCL should be tested in future studies. PMID- 20002357 TI - Invasive zygomycosis in patients with graft-versus-host disease after allogeneic stem cell transplantation. AB - Invasive mold infections are a threat to immunosuppressed patients such as patients with graft-versus-host disease (GVHD) after allogeneic stem cell transplantation (SCT). Up to 10% of SCT recipients develop invasive aspergillosis (IA). Invasive zygomycosis (IZ) may occur during treatment against IA. Here we report 4 SCT patients with GVHD diagnosed with IZ. All patients had received myeloablative hematopoietic SCT and developed chronic GVHD requiring systemic immunosuppression. Underlying diseases were acute lymphocytic leukemia (2), osteomyelofibrosis, and multiple myeloma. All patients had developed pulmonary infiltration that led to initiation of antifungal therapy. Treatment for IA was voriconazole, caspofungin, or itraconazole. Organs involved with zygomycosis were lung, nasal sinus, skin, and kidney. Treatment with liposomal amphotericin and posaconazole was initiated in all patients, and 2 patients also had surgical debridement as well. Despite intensive treatment, no patient survived. IZ is becoming more common in patients with GVHD on successful treatment for IA. Even non-specific symptoms are suspicious in this group of patients and need to be evaluated by vigorous diagnostics. Despite effective antifungals and surgical intervention, the prognosis is grim in patients with active GVHD, as immunoreconstitution is mandatory for successful management. PMID- 20002358 TI - Unusual presentation of chromoblastomycosis due to Cladophialophora carrionii in a renal and pancreas transplant recipient patient successfully treated with posaconazole and surgical excision. AB - Chromoblastomycosis is a chronic, tropical and subtropical, subcutaneous mycosis caused by inoculation of dematiaceous molds. This disease is uncommonly reported in patients who have undergone solid organ transplantation. We describe a case of chromoblastomycosis caused by Cladophialophora carrionii that occurred 7 years after transplantation in a 58-year-old male renal and pancreatic transplant recipient. Diagnosis was based on histopathology and isolation of multiple colonies of the dematiaceous mold in pure culture. Identification was achieved by sequencing of the internal transcribed spacer regions of the rRNA. The patient was successfully treated with posaconazole and surgical excision of a residual lesion. PMID- 20002359 TI - Long-term outcome following liver transplantation for paracetamol overdose. AB - Paracetamol overdose (POD) is a major cause of acute liver failure (ALF) requiring liver transplantation in the United Kingdom. To characterize the early and late outcome after orthotopic liver transplantation (OLT) for POD in the Scottish Liver Transplant Unit over a 14-year period (1992-2006). Data were obtained from a prospective database combined with case-note review. Of 127 liver transplants performed for ALF, 44 were for POD. The median age was 30 (range 18 51). In 18 patients (63.7%), POD was associated with alcohol/other drugs, nine (20.5%) had a staggered overdose and four patients (9.1%) accidentally overdosed. Nineteen patients (43.2%) had a history of previous overdose/psychiatric illness. Post-transplant mortality during the index admission was 30% (13 patients), whilst five patients died during follow-up. The actuarial 5-year patient survival was 54.5%, whilst graft survival was 49.5%. Some 23% of the patients were re transplanted: primary nonfunction (1), hepatic artery thrombosis (3) and chronic rejection (2). Three patients had a subsequent transplant; three patients had two further transplants. Nine patients (35%) continue to have social/psychiatric issues. OLT for POD is associated with significant early and late morbidity and mortality. A multidisciplinary approach is required to identify the suitable candidates, in whom transplantation should be pursued promptly. PMID- 20002360 TI - Functional intravascular volume deficit in patients before surgery. AB - BACKGROUND: Stroke volume (SV) maximization with a colloid infusion, referred to as individualized goal-directed therapy, improves outcome in high-risk surgery. The fraction of patients who need intravascular volume to establish a maximal SV has, however, not been evaluated, and there are only limited data on the volume required to establish a maximal SV before the start of surgery. Therefore, we estimated the occurrence and size of the potential functional intravascular volume deficit in surgical patients. METHODS: Patients scheduled for mastectomy (n=20), open radical prostatectomy (n=20), or open major abdominal surgery (n=20) were anaesthetized, and before the start of surgery, a 200 ml colloid fluid challenge was provided and repeated if a >or=10% increment in SV estimated by oesophageal Doppler was established. The volume needed for SV maximization defined the intravascular volume deficit. RESULTS: Forty-two (70%) of the patients needed volume to establish a maximal SV. For the patients needing volume, the required amount was median 200 ml (range 200-600 ml), with no significant difference between the three groups of patients. The required volume was >or=400 ml in nine patients (15%). CONCLUSION: The majority of anaesthetized patients present with a functional intravascular volume deficit before surgery. Although the deficit in general was minor, a fraction of patients presented with a deficit that may be of clinical relevance, emphasizing the importance of the individual approach of goal-directed fluid therapy. PMID- 20002361 TI - Pain on injection from propofol may be avoided by changing its formulation. AB - BACKGROUND: After using propofol for a decade, pain on injection had been considered routine by patients and medical personnel. When given propofol from a different manufacturer, patients did not complain. Two preparations of propofol were compared. METHODS: A comparative, double-blind, randomized study was conducted in 22 adult patients undergoing pain relief procedures; they received sedation by an intravenous injection of 1.7 mg/kg of propofol and then were treated with paravertebral injections. Pain on injection was assessed by verbal complaint, movement of the extremity, of the whole body and recollection of pain at induction, when discharged. Propofol from Baxter Laboratories, mixed with either 5 ml of 2% lidocaine or 5 ml of NaCl 0.9%, was compared with propofol Laboratorios Gray, which was similarly mixed. Injections were randomly administered four times, blindly, to each of 22 patients. Statistical analysis was conducted using the analysis of variance method. RESULTS: A total of 352 propofol injections were given. Each of the four propofol solutions was administered 88 times; of patients receiving Baxter propofol+saline, 74 (84%) had pain; when mixed with 2% lidocaine 45 (50.2%) complained. After propofol Gray with NaCl 0.9% was given, two patients (2.2%) experienced pain. Propofol Gray with 2% lidocaine produced no pain. None of the latter group remembered having pain, whereas, those given propofol Baxter 54 (61.3%) and 26 (29.5%) remembered experiencing pain at injection. Pain on injection was prevented and statistically reduced (<0.01) with the propofol from Laboratorios Gray. CONCLUSIONS: By changing the formulation (size of molecules and their dispersion) of propofol, pain on injection was avoided. PMID- 20002362 TI - The Richmond Agitation-Sedation Scale: translation and reliability testing in a Swedish intensive care unit. AB - BACKGROUND: Awareness about adequate sedation in mechanically ventilated patients has increased in recent years. The use of a sedation scale to continually evaluate the patient's response to sedation may promote earlier extubation and may subsequently have a positive effect on the length of stay in the intensive care unit (ICU). The Richmond Agitation-Sedation Scale (RASS) provides 10 well defined levels divided into two different segments, including criteria for levels of sedation and agitation. Previous studies of the RASS have shown it to have strong reliability and validity. The aim of this study was to translate the RASS into Swedish and to test the inter-rater reliability of the scale in a Swedish ICU. METHODS: A translation of the RASS from English into Swedish was carried out, including back-translation, critical review and pilot testing. The inter rater reliability testing was conducted in a general ICU at a university hospital in the south of Sweden, including 15 patients mechanically ventilated and sedated. Forty in-pair assessments using the Swedish version of the RASS were performed and the inter-rater reliability was tested using weighted kappa statistics (linear weighting). RESULT: The translation of the RASS was successful and the Swedish version was found to be satisfactory and applicable in the ICU. When tested for inter-rater reliability, the weighed kappa value was 0.86. CONCLUSION: This study indicates that the Swedish version of the RASS is applicable with good inter-rater reliability, suggesting that the RASS can be useful for sedation assessment of patients mechanically ventilated in Swedish general ICUs. PMID- 20002363 TI - Associations of polymorphisms in the promoter I of bovine acetyl-CoA carboxylase alpha gene with beef fatty acid composition. AB - The objectives of this study were to identify single nucleotide polymorphisms (SNPs) in the promoter I (PI) region of the bovine acetyl-CoA carboxylase-alpha (ACACA) gene and to evaluate the extent to which they were associated with lipid related traits. Eight novel SNPs were identified, which were AJ276223:g.2064T>A (SNP1), g.2155C>T (SNP2), g.2203G>T (SNP3), g.2268T>C (SNP4), g.2274G>A (SNP5), g.2340A>G (SNP6), g.2350T>C (SNP7) and g.2370A>G (SNP8). Complete linkage disequilibrium was observed among SNP1, 2, 4, 5, 6 and 8. Phenotypic data were collected from 573 cross-bred steers with six sire breeds, including Hereford, Angus, Brangus, Beefmaster, Bonsmara and Romosinuano. The genotypes of SNP1/2/4/5/6/8 were significantly associated with adjusted backfat thickness. The genotypes of SNP3 were significantly associated with triacylglycerol (TAG) content and fatty acid composition of longissimus dorsi muscle (LM) in Brangus-, Romosinuano- and Bonsmara-sired cattle. Cattle with g.2203GG genotype had greater concentrations of TAG, total lipid, total saturated fatty acid and total monounsaturated fatty acid than did cattle with g.2203GT genotype. The genotypes of SNP7 were significantly associated with fatty acid composition of LM. Cattle with genotype g.2350TC had greater amounts of several fatty acids in LM than did cattle with genotype g.2350CC. Our results suggested that the SNPs in the PI region of ACACA gene are associated with variations in the fatty acid contents in LM. PMID- 20002364 TI - The value of lung ultrasound monitoring in H1N1 acute respiratory distress syndrome. AB - We present the case of a healthy young male who developed acute respiratory failure as a result of infection with influenza A/H1N1 of swine-origin and in whom ventilatory support was optimised and recovery of lung function was monitored by the use of sequential chest ultrasound examinations. The potential pivotal role of bedside lung ultrasonography in H1N1-induced respiratory failure is discussed. PMID- 20002366 TI - Meeting the challenge of developing and maintaining radical hysterectomy skills. PMID- 20002367 TI - Surgical training in gastrointestinal procedures within a UK gynaecological oncology subspecialty programme. AB - OBJECTIVE: An analysis of surgical experience in gastrointestinal procedures within a UK-based gynaecological oncology centre to which subspecialty fellows within the subject are exposed. DESIGN: Retrospective study. SETTING: Northern Gynaecological Oncology Centre, Gateshead, UK. POPULATION: All women undergoing bowel surgery over a six-year period, 1 January 2000 to 31 December 2005. METHODS: Cases were analysed by specialty and grade of surgeon performing the procedure. MAIN OUTCOME MEASURE: Proportion of cases to which subspecialty fellows were exposed. RESULTS: Two hundred and sixty-two women (11.5%) underwent bowel surgery out of 2280 women undergoing major surgery for gynaecological cancer. This included ovarian/primary peritoneal cancer in 186 women (71%). Of these 262 cases, 238 operations (91%) were performed by a gynaecological oncologist, 20 (7.5%) were performed jointly with the gastrointestinal surgeons and four (1.5%) were performed solely by the gastrointestinal surgeons. A gynaecological oncology subspecialty fellow performed 21 (8%) and assisted in an additional 204 operations (78%). Perioperative morbidity and mortality statistics in addition to overall survival outcomes were comparable to the published literature. CONCLUSIONS: A significant proportion of major surgical operations performed within a gynaecological oncology centre require gastrointestinal procedures. The majority of these procedures can be performed by gynaecological oncologists with an acceptable perioperative morbidity and mortality rate. Subspecialty training has the potential to allow trainees significant exposure to these procedures. An accredited post-Fellowship Training Programme can provide the opportunity for hands-on experience to allow gynaecological oncologists the confidence and credibility to perform these procedures independently. PMID- 20002368 TI - Impact of bowel obstruction at the time of initial presentation in women with ovarian cancer. AB - OBJECTIVE: To determine whether the presence of bowel obstruction at the time of initial presentation has any prognostic significance in these women. DESIGN: Retrospective cohort study. SETTING: Dedicated gynaecological oncology service of a large tertiary institution. POPULATION: Women who had a bowel obstruction as part of their initial presentation of ovarian cancer were identified between 1995 and 2007. Each woman was matched with four control women (with disease but no obstruction). METHODS: Women with disease were compared with controls to determine the impact, if any, of bowel obstruction at presentation. Several prognostic variables including bowel obstruction were also evaluated in a Cox proportional hazard model. MAIN OUTCOME MEASURES: Progression-free survival (PFS) and overall survival (OS). RESULTS: Forty-eight women with disease and 192 controls were identified during the study period. The median follow-up period was 19 months among women with disease versus 20 months in controls. No differences were seen in demographics and clinical characteristics of the women. Optimal cytoreduction rate was similar between the two groups (75% versus 78%, P = 0.7). Patients with bowel obstruction had a shorter PFS and OS compared with controls [19 months versus 21 months (P = 0.01) and 22 versus 35 months (P = 0.008)], respectively. Bowel obstruction at presentation was an independent prognostic variable with a hazard ratio of 1.5 (P = 0.009). Other prognostic variables were age, stage and extent of surgical cytoreduction. CONCLUSIONS: Bowel obstruction at the time of initial presentation is an adverse prognostic factor in women with ovarian cancer. PMID- 20002370 TI - Laparoscopic hysteropexy: the initial results of a uterine suspension procedure for uterovaginal prolapse. AB - OBJECTIVE: The aim of this study was to evaluate the outcome of laparoscopic hysteropexy, a surgical technique for the management of uterine prolapse, involving suspension of the uterus from the sacral promontory using bifurcated polypropylene mesh. DESIGN: The investigation was designed as a prospective observational study (clinical audit). SETTING: The study was undertaken at a tertiary referral urogynaecology unit in the UK. POPULATION: The participants comprised 51 consecutive women with uterovaginal prolapse, who chose laparoscopic hysteropexy as one of the available surgical options. METHODS: The hysteropexy was conducted laparoscopically in all cases. A bifurcated polypropylene mesh was used to suspend the uterus from the sacral promontory. The two arms of the mesh were introduced through bilateral windows created in the broad ligaments, and were sutured to the anterior cervix; the mesh was then fixed to the anterior longitudinal ligament over the sacral promontory, to elevate the uterus. MAIN OUTCOME MEASURES: Cure of the uterine prolapse was evaluated subjectively using the International Consultation on Incontinence Questionnaire for vaginal symptoms (ICIQ-VS), and objectively by vaginal examination using the Baden-Walker halfway system and the pelvic organ prolapse quantification (POP-Q) scale. Operative and postoperative complications were also assessed. RESULTS: The mean age of the 51 women was 52.5 years (range 19-71 years). All were sexually active, and at least three of them expressed a strong desire to have children in the future. All were available for follow-up in clinic at 10 weeks, and 38 have completed the questionnaires. In 50 out of 51 women the procedure was successful, with no objective evidence of uterine prolapse on examination at follow-up; there was one failure. Significant subjective improvements in prolapse symptoms, sexual wellbeing and related quality of life were observed, as detected by substantial reductions in the respective questionnaire scores. CONCLUSIONS: Laparoscopic hysteropexy is both a feasible and an effective procedure for correcting uterine prolapse without recourse to hysterectomy. It allows restoration of the length of the vagina without compromising its calibre, and is therefore likely to have a favourable functional outcome. PMID- 20002369 TI - A combination of misoprostol and estradiol for preoperative cervical ripening in postmenopausal women: a randomised controlled trial. AB - OBJECTIVE: To compare the impact of 1000 microg of self-administered vaginal misoprostol versus self-administered vaginal placebo on preoperative cervical ripening after 2 weeks of pretreatment with estradiol vaginal tablets in postmenopausal women prior to day-care operative hysteroscopy. DESIGN: Randomised, double-blind, placebo-controlled sequential trial. SETTING: Norwegian university teaching hospital. POPULATION: Sixty-seven postmenopausal women referred for day-care operative hysteroscopy. METHODS: The women were randomised to receive either 1000 microg of self-administered vaginal misoprostol or self administered vaginal placebo on the evening before day-care operative hysteroscopy. All women had administered a 25-microg vaginal estradiol tablet daily for 14 days prior to the operation. MAIN OUTCOME MEASURES: PRIMARY OUTCOME: preoperative cervical dilatation at hysteroscopy. SECONDARY OUTCOMES: difference in dilatation at recruitment and before hysteroscopy, number of women who achieved a preoperative cervical dilatation of 5 mm or more, acceptability, complications and adverse effects. RESULTS: The mean cervical dilatation was 5.7 mm (SD, 1.6 mm) in the misoprostol group and 4.7 mm (SD, 1.5 mm) in the placebo group, the mean difference in cervical dilatation being 1.0 mm (95% CI, 0.2-1.7 mm). Self-administered vaginal misoprostol of 1000 microg at home on the evening before day-care hysteroscopy is safe and highly acceptable, although a small proportion of women experienced lower abdominal pain. CONCLUSIONS: One thousand micrograms of self-administered vaginal misoprostol, 12 hours prior to day-care hysteroscopy, after 14 days of pretreatment with vaginal estradiol, has a significant cervical ripening effect compared with placebo in postmenopausal women. PMID- 20002371 TI - Estimating the risk of gestational diabetes mellitus: a clinical prediction model based on patient characteristics and medical history. AB - OBJECTIVE: To develop a clinical prediction rule that can help the clinician to identify women at high and low risk for gestational diabetes mellitus (GDM) early in pregnancy in order to improve the efficiency of GDM screening. DESIGN: We used data from a prospective cohort study to develop the clinical prediction rule. SETTING: The original cohort study was conducted in a university hospital in the Netherlands. POPULATION: Nine hundred and ninety-five consecutive pregnant women underwent screening for GDM. METHODS: Using multiple logistic regression analysis, we constructed a model to estimate the probability of development of GDM from the medical history and patient characteristics. Receiver operating characteristics analysis and calibration were used to assess the accuracy of the model. MAIN OUTCOME MEASURE: The development of a clinical prediction rule for GDM. We also evaluated the potential of the prediction rule to improve the efficiency of GDM screening. RESULTS: The probability of the development of GDM could be predicted from the ethnicity, family history, history of GDM and body mass index. The model had an area under the receiver operating characteristic curve of 0.77 (95% CI 0.69-0.85) and calibration was good (Hosmer and Lemeshow test statistic, P = 0.25). If an oral glucose tolerance test was performed in all women with a predicted probability of 2% or more, 43% of all women would be tested and 75% of the women with GDM would be identified. CONCLUSIONS: The use of a clinical prediction model is an accurate method to identify women at increased risk for GDM, and could be used to select women for additional testing for GDM. PMID- 20002372 TI - Toll-like receptors 2 and 4 and the cryopyrin inflammasome in normal pregnancy and pre-eclampsia. AB - OBJECTIVE: Pre-eclampsia involves a maternal inflammatory response that differs from both normal pregnancy and normotensive intrauterine growth restriction (IUGR). Our objective was to examine neutrophil Toll-like receptor (TLR), cryopyrin, nuclear factor-kappaB (NF-kappaB) subunit and interleukin-1beta (IL 1beta), and inflammatory cytokine profiles in women with pre-eclampsia or normotensive IUGR, as well as in normal pregnancy and non-pregnancy controls. DESIGN AND METHOD: A case-control study was performed. We examined the messenger RNA (mRNA) and protein expressions of TLR4 and TLR2, mRNA levels of cryopyrin, IL 1beta, NF-kappaB subunits p50 and p65, as well as maternal serum inflammatory cytokine profiles (IL-2, IL-6, tumour necrosis factor-alpha [TNF-alpha], interferon-gamma [IFN-gamma] and IL-10) in women with and without pre-eclampsia using real-time reverse transcription polymerase chain reactions, flow cytometry and multiplex immunoassays. SETTING: A single tertiary maternity hospital in Vancouver, Canada. POPULATION: Women with early-onset pre-eclampsia (<34 weeks of gestation, n = 25), women with late-onset pre-eclampsia (>or=34(+0) weeks of gestation, n = 25), women with normotensive IUGR (n = 25), women with normal pregnancy (n = 75) and non-pregnancy (n = 25) controls. RESULTS: Women with pre eclampsia (as a single combined group of early- and late-onset, and particularly in women with early-onset pre-eclampsia) had increased TLR2 and TLR4 mRNA and protein expressions elevated cryopyrin, NF-kappaB subunit, and IL-1beta mRNA expression, and TNF-alpha:IL-10 and IL-6:IL-10 ratios compared with other groups. CONCLUSIONS: These data suggest that TLRs and cryopyrin may modulate the innate immune response of the maternal syndrome of pre-eclampsia, and might also trigger the differential inflammatory response existing between early onset pre-eclampsia and normotensive IUGR. PMID- 20002373 TI - Defining the surgical management of suspected early-stage ovarian cancer by estimating patient numbers through alternative management strategies. PMID- 20002376 TI - Vesicovaginal fistula and pregnancy. PMID- 20002378 TI - Gum chewing stimulates early return of bowel motility after caesarean section. PMID- 20002379 TI - Oxytocin at caesarean section - are we giving too much? PMID- 20002381 TI - The role of bacterial vaginosis, aerobic vaginitis, abnormal vaginal flora and the risk of preterm birth. PMID- 20002383 TI - Inter-observer agreement in clinical decision-making for abnormal cardiotocogram during labour: a comparison between CTG and CTG plus STAN. PMID- 20002385 TI - Cell salvage at caesarean section: the need for an evidence-based approach. PMID- 20002389 TI - Flagellar oscillation: a commentary on proposed mechanisms. AB - Eukaryotic flagella and cilia have a remarkably uniform internal 'engine' known as the '9+2' axoneme. With few exceptions, the function of cilia and flagella is to beat rhythmically and set up relative motion between themselves and the liquid that surrounds them. The molecular basis of axonemal movement is understood in considerable detail, with the exception of the mechanism that provides its rhythmical or oscillatory quality. Some kind of repetitive 'switching' event is assumed to occur; there are several proposals regarding the nature of the 'switch' and how it might operate. Herein I first summarise all the factors known to influence the rate of the oscillation (the beating frequency). Many of these factors exert their effect through modulating the mean sliding velocity between the nine doublet microtubules of the axoneme, this velocity being the determinant of bend growth rate and bend propagation rate. Then I explain six proposed mechanisms for flagellar oscillation and review the evidence on which they are based. Finally, I attempt to derive an economical synthesis, drawing for preference on experimental research that has been minimally disruptive of the intricate structure of the axoneme. The 'provisional synthesis' is that flagellar oscillation emerges from an effect of passive sliding direction on the dynein arms. Sliding in one direction facilitates force-generating cycles and dynein-to dynein synchronisation along a doublet; sliding in the other direction is inhibitory. The direction of the initial passive sliding normally oscillates because it is controlled hydrodynamically through the alternating direction of the propulsive thrust. However, in the absence of such regulation, there can be a perpetual, mechanical self-triggering through a reversal of sliding direction due to the recoil of elastic structures that deform as a response to the prior active sliding. This provisional synthesis may be a useful basis for further examination of the problem. PMID- 20002390 TI - To speciate, or not to speciate? Resource heterogeneity, the subjectivity of similarity, and the macroevolutionary consequences of niche-width shifts in plant feeding insects. AB - Coevolutionary studies on plants and plant-feeding insects have significantly improved our understanding of the role of niche shifts in the generation of new species. Evolving plant lineages essentially constitute moving islands and archipelagoes in resource space, and host shifts by insects are usually preceded by colonizations of novel resources. Critical to hypotheses concerning ecological speciation is what happens immediately before and after colonization attempts: if an available plant is too similar to the current host(s), it simply will be incorporated into the existing diet, but if it is too different, it will not be colonized in the first place. It thus seems that the probability of speciation is maximized when alternative hosts are at an 'intermediate' distance in resource space. In this review, I wish to highlight the possibility that resource similarity and, thus, the definition of 'intermediate', are subjective concepts that depend on the herbivore lineage's tolerance to dietary variation. This subjectivity of similarity means that changes in tolerance can either decrease or increase speciation probabilities depending on the distribution of plants in resource space: insect lineages with narrow tolerances are likely to speciate by 'island-hopping' on young, species-rich plant groups, whereas more generalized lineages could speciate by shifting among resource archipelagoes formed by higher plant taxa. Repeated and convergent origins of traits known to broaden or to restrict host-plant use in multiple different insect groups provide opportunities for studying how tolerance and resource heterogeneity may interact to determine speciation rates. PMID- 20002391 TI - Global interrelationships of Plesiosauria (Reptilia, Sauropterygia) and the pivotal role of taxon sampling in determining the outcome of phylogenetic analyses. AB - Previous attempts to resolve plesiosaurian phylogeny are reviewed and a new phylogenetic data set of 66 taxa (67% of ingroup taxa examined directly) and 178 characters (eight new) is presented. We recover two key novel results: a monophyletic Plesiosauridae comprising Plesiosaurus dolichodeirus, Hydrorion brachypterygius, Microcleidus homalospondylus, Occitanosaurus tournemirensis and Seeleyosaurus guilelmiimperatoris; and five plesiosaurian taxa recovered outside the split between Plesiosauroidea and Pliosauroidea. These taxa are Attenborosaurus conybeari, 'Plesiosaurus'macrocephalus and a clade comprising Archaeonectrus rostratus, Macroplata tenuiceps and BMNH 49202. Based on this result, a new name, Neoplesiosauria, is erected for the clade comprising Plesiosauroidea and Pliosauroidea. Taxon subsamples of the new dataset are used to simulate previous investigations of global plesiosaurian relationships. Based on these simulations, most major differences between previous global phylogenetic hypotheses can be attributed to differences in taxon sampling. These include the position of Leptocleididae and Polycotylidae and the monophyly or paraphyly of Rhomaleosauridae. On this basis we favour the results recovered by our, larger analysis. Leptocleididae and Polycotylidae are sister taxa, forming a monophyletic clade within Plesiosauroidea, indicating that the large-headed, short-necked 'pliosauromorph' body plan evolved twice within Plesiosauria. Rhomaleosauridae forms the monophyletic sister taxon of Pliosauridae within Pliosauroidea. Problems are identified with previous phylogenetic definitions of plesiosaurian clades and new, stem-based definitions are presented that should maintain their integrity over a range of phylogenetic hypotheses. New, rank-free clade names Cryptoclidia and Leptocleidia are erected to replace the superfamilies Cryptoclidoidea and Leptocleidoidea. These were problematic as they were nested within the superfamily Plesiosauroidea. The incongruence length difference test indicates no significant difference in levels of homoplasy between cranial and postcranial characters. PMID- 20002392 TI - Adolescent reproductive health. Editor's choice. PMID- 20002393 TI - Lost in transition: the needs of adolescents with Turner syndrome. PMID- 20002394 TI - Hyperandrogenaemia in adolescent girls: origins of abnormal gonadotropin releasing hormone secretion. PMID- 20002395 TI - Reproductive endocrinology of adolescent polycystic ovary syndrome. AB - Polycystic ovary syndrome (PCOS) is the most common endocrinopathy among reproductive-aged women, and it typically presents during adolescence. The objective of this review is to describe the clinical manifestations of PCOS in adolescent girls and the underlying basis for the altered reproductive physiology. Recognising adolescents at risk for PCOS and taking the appropriate steps to reduce circulating androgen levels is critical in reducing the clinical symptomatology of this disorder, and the development of adulthood infertility, diabetes, and metabolic syndrome in patients with PCOS. PMID- 20002396 TI - The histology and management of ovarian cysts found in children and adolescents presenting to a children's hospital from 1991 to 2007: a call for more paediatric gynaecologists. AB - OBJECTIVE: To determine the nature and surgical management of ovarian cysts. DESIGN: Retrospective case-note study. SETTING: Large Childrens' Hospital: Alder Hey NHS Foundation Trust. SAMPLE: Children undergoing surgery for ovarian cysts between 1991 and 2007. METHODS: Cases identified using coding and the pathology database, and analysed using snap 9. MAIN OUTCOME MEASURES: Extent of surgery performed. Size and histological features of cysts removed. RESULTS: A total of 155 cases were identified. There were 62 ovarian cysts in children under nine who were prepubertal. There were 58 neoplastic cysts in total. Most were benign teratomas (36). Ten cysts were malignant, including five granulosa cell tumours, one yolk sac tumour, one endodermal sinus tumour and one dysgerminoma. Tumour markers were performed in only 16 cases (10%). Sixty-one (39%) had an ultrasound scan and 16 (10%) had a computed tomography (CT) or magnetic resonance imaging (MRI) scan. Ninety girls (58%) had an oophorectomy and 40 (25%) had an ovarian cystectomy. Oophorectomies were performed for all cases of malignancy, but 75 were also performed for benign or normal pathology. Only 16% of cases were referred to the paediatric gynaecologist and all were for post-operative management. CONCLUSIONS: We recommend the greater use of imaging of the pelvis and tumour markers preoperatively. There should be greater use of conservative expectant management or ovarian-sparing surgery in view of the low risk of malignancy in this age group. The practice of removing ovaries for benign cysts may be overcome by appointing more gynaecologists with advanced training skills training in paediatric and adolescent gynaecology. PMID- 20002397 TI - The clandestine uterus: or how the uterus escapes detection prior to puberty. AB - Uterine agenesis is one of the differential diagnoses in adolescent girls with delayed menstruation. It may also be suspected earlier in childhood during investigations for other genitourinary conditions. However, accurate confirmation that the uterus is absent can be extremely difficult before puberty because of its small size. We describe ten girls referred to a specialist centre with a presumed diagnosis of an absent uterus which was later found to be incorrect. We conclude that imaging should be undertaken by clinicians with experience in management of this age group and in some girls it may be necessary to delay final diagnosis until after puberty. PMID- 20002398 TI - Oocyte cryopreservation after controlled ovarian hyperstimulation in mosaic Turner syndrome: another fertility preservation option in a dedicated UK clinic. PMID- 20002399 TI - Conservation of fertility and oocyte genetics in a young woman with mosaic Turner syndrome. PMID- 20002400 TI - Modeling familial association of ages at onset of disease in the presence of competing risk. AB - In genetic family studies, ages at onset of diseases are routinely collected. Often one is interested in assessing the familial association of ages at the onset of a certain disease type. However, when a competing risk is present and is related to the disease of interest, the usual measure of association by treating the competing event as an independent censoring event is biased. We propose a bivariate model that incorporates two types of association: one is between the first event time of paired members, and the other is between the failure types given the first event time. We consider flexible measures for both types of association, and estimate the corresponding association parameters by adopting the two-stage estimation of Shih and Louis (1995, Biometrics 51, 1384-1399) and Nan et al. (2006, Journal of the American Statistical Association 101, 65-77). The proposed method is illustrated using the kinship data from the Washington Ashkenazi Study. PMID- 20002401 TI - Small-sample estimation of species richness applied to forest communities. AB - Many well-known methods are available for estimating the number of species in a forest community. However, most existing methods result in considerable negative bias in applications, where field surveys typically represent only a small fraction of sampled communities. This article develops a new method based on sampling with replacement to estimate species richness via the generalized jackknife procedure. The proposed estimator yields small bias and reasonably accurate interval estimation even with small samples. The performance of the proposed estimator is compared with several typical estimators via simulation study using two complete census datasets from Panama and Malaysia. PMID- 20002402 TI - Sequential analysis of longitudinal data in a prospective nested case-control study. AB - The nested case-control design is a relatively new type of observational study whereby a case-control approach is employed within an established cohort. In this design, we observe cases and controls longitudinally by sampling all cases whenever they occur but controls at certain time points. Controls can be obtained at time points randomly scheduled or prefixed for operational convenience. This design with longitudinal observations is efficient in terms of cost and duration, especially when the disease is rare and the assessment of exposure levels is difficult. In our design, we propose sequential sampling methods and study both (group) sequential testing and estimation methods so that the study can be stopped as soon as the stopping rule is satisfied. To make such a longitudinal sampling more efficient in terms of both numbers of subjects and replications, we propose applying sequential sampling methods to subjects and replications, simultaneously, until the information criterion is fulfilled. This simultaneous sequential sampling on subjects and replicates is more flexible for practitioners designing their sampling schemes, and is different from the classical approaches used in longitudinal studies. We newly define the sigma-field to accommodate our proposed sampling scheme, which contains mixtures of independent and correlated observations, and prove the asymptotic optimality of sequential estimation based on the martingale theories. We also prove that the independent increment structure is retained so that the group sequential method is applicable. Finally, we present results by employing sequential estimation and group sequential testing on both simulated data and real data on children's diarrhea. PMID- 20002403 TI - Design and analysis of multiple events case-control studies. AB - In case-control research where there are multiple case groups, standard analyses fail to make use of all available information. Multiple events case-control (MECC) studies provide a new approach to sampling from a cohort and are useful when it is desired to study multiple types of events in the cohort. In this design, subjects in the cohort who develop any event of interest are sampled, as well as a fraction of the remaining subjects. We show that a simple case-control analysis of data arising from MECC studies is biased and develop three general estimating-equation-based approaches to analyzing data from these studies. We conduct simulation studies to compare the efficiency of the various MECC analyses with each other and with the corresponding conventional analyses. It is shown that the gain in efficiency by using the new design is substantial in many situations. We demonstrate the application of our approach to a nested case control study of the effect of oral sodium phosphate use on chronic kidney injury with multiple case definitions. PMID- 20002404 TI - Regret-regression for optimal dynamic treatment regimes. AB - We consider optimal dynamic treatment regime determination in practice. Model building, checking, and comparison have had little or no attention so far in this literature. Motivated by an application on optimal dosage of anticoagulants, we propose a modeling and estimation strategy that incorporates the regret functions of Murphy (2003, Journal of the Royal Statistical Society, Series B 65, 331-366) into a regression model for observed responses. Estimation is quick and diagnostics are available, meaning a variety of candidate models can be compared. The method is illustrated using simulation and the anticoagulation application. PMID- 20002418 TI - "Thinking outside the box" in maternity care. PMID- 20002419 TI - Nurse-midwives' experiences with planned home birth: impact on attitudes and practice. AB - BACKGROUND: Health care providers' attitudes toward maternity care options influence the nature of informed decision-making discussions and patient choice. A woman's choice of birth site may be affected by her provider's opinion and practice site. The objectives of this study were to describe American nurse midwives' attitudes toward, and experiences with, planned home birth, and to explore correlates and predictors of their attitudes toward planned home birth as measured by the Provider Attitudes towards Planned Home Birth (PAPHB) scale. METHODS: A survey instrument, which incorporates the PAPHB and assesses demographic, education, practice, personal experience, and external barrier variables that may predict attitudes toward planned home birth practice, was completed by 1,893 nurse-midwives. Bivariate analysis identified associations between variables and attitudes. Linear regression modeling identified predictors of attitudes. RESULTS: Variables that significantly predicted favorable attitudes to planned home birth were increased clinical and educational experiences with planned home birth (p < 0.001), increased exposure to planned home birth (p < 0.001), and younger age (p < 0.001). External barriers that significantly predicted less favorable attitudes included financial (p = 0.03) and time (p < 0.001) constraints, inability to access medical consultation (p < 0.001), and fear of peer censure (p < 0.001). Willingness to practice in the home was correlated with factors related to nurse-midwives' confidence in their management abilities and beliefs about planned home birth safety. CONCLUSIONS: The results suggest that nurse-midwives' choice of practice site and comfort with planned home birth are strongly influenced by the nature and amount of exposure to home birth during professional education or practice experiences, in addition to interprofessional, logistic, and environmental factors. Findings from this research may inform interdisciplinary education and collaborative practice in the area of planned home birth. PMID- 20002420 TI - Postpartum perineal pain in a low episiotomy setting: association with severity of genital trauma, labor care, and birth variables. AB - BACKGROUND: Perineal pain is common after childbirth. We studied the effect of genital tract trauma, labor care, and birth variables on the incidence of pain in a population of healthy women exposed to low rates of episiotomy and operative vaginal delivery. METHODS: A prospective study of genital trauma at birth and assessment of postpartum perineal pain and analgesic use was conducted in 565 midwifery patients. Perineal pain was assessed using the present pain intensity (PPI) and visual analog scale (VAS) components of the validated short-form McGill pain scale. Multivariate logistic regression examined which patient characteristics or labor care measures were significant determinants of perineal pain and use of analgesic medicines. RESULTS: At hospital discharge, women with major trauma reported higher VAS pain scores (2.16 +/- 1.61 vs 1.48 +/- 1.40; p < 0.001) and were more likely to use analgesic medicines (76.3 vs 23.7%, p = 0.002) than women with minor or no trauma. By 3 months, average VAS scores were low in each group and not significantly different. Perineal pain at the time of discharge was associated in univariate analysis with higher education level, ethnicity (non-Hispanic white), nulliparity, and longer length of active maternal pushing efforts. In a multivariate model, only trauma group and length of active pushing predicted the pain at hospital discharge. In women with minor or no trauma, only length of the active part of second stage labor had a positive relationship with pain. In women with major trauma, the length of active second stage labor had no independent effect on the level of pain at discharge beyond its effect on the incidence of major trauma. CONCLUSIONS: Women with spontaneous perineal trauma reported very low rates of postpartum perineal pain. Women with major trauma reported increased perineal pain compared with women who had no or minor trauma; however, by 3 months postpartum this difference was no longer present. In women with minor or no perineal trauma, a longer period of active pushing was associated with increased perineal pain. PMID- 20002421 TI - A birth education program for expectant fathers in Taiwan: effects on their anxiety. AB - BACKGROUND: A father who does not know how to assist the mother in relieving labor pains may experience a sense of powerlessness and anxiety. The objective of this study was to evaluate how an education program for expectant fathers who attended their partners' labor and birth affected their anxiety. METHODS: In a randomized controlled trial, 87 expectant fathers who attended their pregnant partners through labor and birth at a hospital in central Taiwan were allocated by block randomization to an experimental (n = 45) and a control (n = 42) group. The men completed their basic personal information, a childbirth expectations questionnaire, and a Trait Anxiety Inventory when they were recruited. Two hours after birth of their child, all the expectant fathers completed a State of Anxiety Inventory. RESULTS: Our results showed no statistically significant differences between the experimental and control groups of fathers in trait anxiety and their prenatal childbirth expectations. After analysis of covariance (ANCOVA) was applied to correct for education level, sources of childbirth information, attendance at Lamaze childbirth classes, and childbirth expectations at baseline, the effect of the childbirth program was significant for the postnatal level of anxiety (F = 3.38, p = 0.001). CONCLUSIONS: The study findings justify the clinical implementation of a birth education program based on the self-efficacy theory as an effective means of reducing anxiety among expectant fathers. PMID- 20002422 TI - The experience of planned home birth: views of the first 500 women. AB - BACKGROUND: Home birth remains a contentious issue in North America. Professional regulatory bodies are in conflict about the safety of home birth as an option for healthy women. The voices of women have largely been ignored in this debate. The purpose of this study is to report on the experiences of 559 women who had a planned home birth over a 2-year period in British Columbia, Canada. METHODS: We asked all women in the Province of British Columbia who had planned for their birth to be at home with a regulated midwife in attendance to answer an open ended question about positive and negative aspects of their birth. The qualitative method of interpretive description was used to understand what women believed to be the essence of their experience. RESULTS: Women felt strongly positive about their trust in their midwife's skill and knowledge, a sense of emotional support and empowerment attained through their relationship with the midwife, perceptions of relaxation in their own home, being informed and included in the planning of their care, and the amount of time the midwife spent with their family. They believed that the confidence arising from their intense preparation and partnership with their midwives permitted them to choreograph their birth experience to a degree that would not be possible in a formal setting. CONCLUSIONS: Women who planned a home birth with a registered midwife in British Columbia were overwhelmingly positive about their experience. Our qualitative report underscores the value women place on having the choice to give birth at home. PMID- 20002423 TI - Men's psychological transition to fatherhood: an analysis of the literature, 1989 2008. AB - BACKGROUND: The most recent review on men's transition to fatherhood was published in 1986. The present paper reports on how the literature has portrayed fatherhood over the past 20 years. The aim was to investigate men's psychological transition to fatherhood from pregnancy of the partner through the infant's first year of life. METHODS: The PsycINFO, PubMed, MEDLINE, Ingenta, Ovid, EMBASE, and WoS databases were accessed to conduct a literature search on the topic. The concepts of self-image transformation, triadic relationship development, and social environment influence were used to examine the complexity of the fatherhood transition process. Specific focus was placed on men's intrapsychic relational and social dimensions. RESULTS: Our analysis of the yielded results revealed three specific fatherhood stages: prenatal, labor and birth, and postnatal periods. Partner pregnancy was found to be the most demanding period in terms of psychological reorganization of the self. Labor and birth were the most intensely emotional moments, and the postnatal period was most influenced by environmental factors. The latter was also experienced as being the most interpersonally and intrapersonally challenging in terms of coping with the new reality of being a father. CONCLUSIONS: Men's transition to fatherhood is guided by the social context in which they live and work and by personal characteristics in interplay with the quality of the partner relationship. Men struggle to reconcile their personal and work-related needs with those of their new families. PMID- 20002424 TI - Top-selling childbirth advice books: a discourse analysis. AB - BACKGROUND: Recent evidence suggests that one-third of women receive information about pregnancy and childbirth through books. Messages about what characteristics are normal (or expected) in childbirth are disseminated in a variety of ways, including popular childbirth education books, but little study of them has been conducted. The purpose of this investigation is to address that gap by examining the discussions about childbirth in the 10 top-selling books in the United States. METHODS: Discourse analysis (relating to the public, personal, and political discussions about a specific phenomenon) was used to study 10 best selling United States childbirth advice books marketed to childbearing women during the first week of November 2007. RESULTS: Book styles ranged from clinical descriptions of pregnancy and birth primarily offering reassurance, self-help information, and danger signs to more folksy and humorous commentaries. Presentation of scientific evidence to support recommendations was uneven and at times inaccurate. Five focal areas of discourse included body image, labor and birth, pain, power and control, and life preparation for motherhood. CONCLUSIONS: Top-selling books shine an interesting light on the current state of United States maternity practices. Women and health professionals should assess them carefully and engage with each other about their recommendations and implications for childbirth. PMID- 20002425 TI - Hurricane Katrina and perinatal health. AB - We review the literature on the effects of Hurricane Katrina on perinatal health, and providing data from our own research on pregnant and postpartum women. After Katrina, obstetric, prenatal, and neonatal care was compromised in the short term, but increases in adverse birth outcomes such as preterm birth, low birthweight, and maternal complications were mostly limited to highly exposed women. Both pregnant and postpartum women had rates of post-traumatic stress disorder similar to, or lower than, others exposed to Katrina, and rates of depression similar to other pregnant and postpartum populations. Health behaviors, such as smoking and breastfeeding, may have been somewhat negatively affected by the disaster, whereas effects on nutrition were likely associated with limited time, money, and food choices, and indicated by both weight gain and loss. We conclude that, with a few specific exceptions, postdisaster concerns and health outcomes for pregnant and postpartum women were similar to those of other people exposed to Hurricane Katrina. In such situations, disaster planners and researchers should focus on providing care and support for the normal concerns of the peripartum period, such as breastfeeding, depression, and smoking cessation. Contraception needs to be available for those who do not want to become pregnant. Although additional physical and mental health care needs to be provided for the most severely exposed women and their babies, many women are capable of surviving and thriving in postdisaster environments. PMID- 20002426 TI - Roundtable discussion: Early labor: what's the problem? AB - PREFACE: In places where hospital birth is the norm, one of the major contemporary challenges to the organization of intrapartum care is posed by women who are not in established labor. In the United Kingdom, these women have been given a special name, "Category X," and they can account for a substantial percentage of admissions (1). These women are not deemed to be in need of hospital care, but the women themselves may feel otherwise as they struggle to understand the sensations they are experiencing. Until relatively recently, little research effort was expended on early and latent phase labor, reflecting, perhaps, the assumption that it is just a gentle and relatively straightforward preamble to the "real thing" that can easily be dealt with by keeping mobile, leaning over furniture, or doing the ironing. Because early labor is not seen as needing a health professional's input, the message is that it is unimportant. However, emerging evidence is challenging that view. Four large randomized controlled trials have recently evaluated interventions related to early labor care (2-5), stimulated by concerns that included repeated visits to the labor ward and the impact of early admission with the potential for a cascade of interventions. These trials, and other research reporting women's own perspectives on labor onset, reflect growing awareness that this stage of labor merits consideration in its own right. An International Early Labor Research Group has formed who will develop the evidence base in this important part of childbearing. The group represents varied disciplines including midwifery, psychology, epidemiology, antenatal education, and service user representatives. Members of this group are among those who have contributed to this Roundtable Discussion. The contributions draw attention to the complexities of early labor and its importance for childbearing women, their caregivers and companions. We might reasonably hypothesize that a woman's experience of early labor sets the scene for what follows, and it is clear that this is an area worthy of considerable further research. PMID- 20002427 TI - BirthStats: Percentage of mothers receiving epidural/spinal anesthesia by age, race, and Hispanic origin of mother: total of 18 U.S. reporting areas, singletons only, 2006. PMID- 20002428 TI - Letter from North America: The United States government's comparative effectiveness research program. PMID- 20002429 TI - Oxytocin as a "high alert medication": a multilayered challenge to the status quo. AB - Oxytocin is the drug most commonly associated with preventable adverse perinatal outcomes. In 2007 it was added to the Institute for Safe Medication Practices short list of medications "bearing a heightened risk of harm," which may "require special safeguards to reduce the risk of error." In January 2009 the American Journal of Obstetrics and Gynecology published a Clinical Opinion paper about oxytocin's inclusion on the list and how the obstetrics profession in the United States should respond. The authors call for the development of specific evidence based guidelines to reduce the likelihood of patient harm by limiting elective use of oxytocin, decreasing the need for indicated use, reducing dosages during necessary use, giving more responsibility and authority for the patient's safety to the professional who is "at the bedside administering and monitoring the oxytocin infusion" (i.e., the nurse), and accepting that "more time rather than more oxytocin is generally preferable" once adequate uterine activity has been achieved. It is unfortunate that this important paper discounted the risk of harm from cesarean sections and did not mention the strong linkage between epidural analgesia and use of oxytocin. Physicians, midwives, nurses, and others should examine and discuss these issues further in view of increased alertness to the risk of harm from unsafe use of oxytocin. PMID- 20002434 TI - "The natural caesarean". PMID- 20002436 TI - Current status of robotic surgery for pelvic organ prolapse. PMID- 20002437 TI - THIS ARTICLE HAS BEEN RETRACTED. Physical therapy for premature ejaculation, erectile dysfunction and chronic pelvic pain syndrome. AB - There is limited knowledge about the exact role of the pelvic floor in male sexual functioning. Pelvic floor muscle function might be involved in the enhancement of blood flow to the penis, and evidence suggests an active role for the ischio- and bulbocavernous muscles and other pelvic floor muscles in the initiation and maintenance of erection. Increased activity of pelvic floor muscles might also be preparatory to ejaculation. Studies have shown positive results after physical therapy for erectile dysfunction, premature ejaculation and chronic pelvic pain syndrome. However, the methodological quality of some of these studies is poor and further research validating specific physical therapies in the assessment and treatment of male sexual function is necessary. In this respect physical therapists have a potential role as integral members of healthcare teams involved in the improvement of male sexual health. PMID- 20002438 TI - Serum sex hormones and the 20-year risk of lower urinary tract symptoms in community-dwelling older men. AB - OBJECTIVE: To evaluate serum sex steroid hormone concentrations and long-term risk of subsequent lower urinary tract symptoms (LUTS) in a cohort of community dwelling older men. SUBJECTS AND METHODS: Between 1984 and 1987, serum sex hormone concentrations were measured in participants in the Rancho Bernardo Study, a prospective, community-based study. In 2006, the American Urological Association Symptom Index (AUA-SI) was mailed to surviving male participants. Logistic regression was used to examine associations of baseline hormone concentrations with AUA-SI. RESULTS: Among 158 surviving men with complete data and no history of prostate cancer, the mean (sd) age at serum sex steroid assessment was 58 (6.6) years with a mean (sd) follow-up of 20.3 (0.6) years. In age-adjusted logistic regression, there was a significant inverse association of testosterone : dihydrotestosterone (DHT) with LUTS (P = 0.05). Also, men with higher concentrations of bioavailable testosterone had a 56% decreased risk of LUTS compared with those with hypogonadal concentrations, although the association was not statistically significant (odds ratios 0.44, 95% confidence interval 0.14-1.40) or distributed evenly among quartiles. There were no significant associations of total testosterone, oestradiol (E(2)), testosterone : E(2), DHT, or dehydroepiandrosterone with LUTS or with any measured hormones and urinary bother. CONCLUSIONS: In this cohort, men with higher mid-life levels of testosterone : DHT and bioavailable testosterone had a decreased 20-year risk of LUTS. These data support other studies reporting inverse associations of serum testosterone with LUTS. Clinical trials of testosterone therapy should include LUTS and clinical benign prostatic hyperplasia as outcomes. PMID- 20002439 TI - Pathological stage review is indicated in primary pT1 bladder cancer. AB - OBJECTIVE: To evaluate the effect of a pathology review on the clinical outcome of patients with primary pT1 bladder cancer (BC), as the clinical course of such patients is variable. PATIENTS AND METHODS: The slides of 164 primary (first diagnosis) pT1 bladder tumours from two university hospitals were reviewed by one pathologist for stage and grade (World Health Organization 1973 and 2004). Patients were initially managed conservatively with bacille Calmette-Guerin (BCG). Uni- and multivariate analyses compared the predictive value of age, gender, hospital, carcinoma in situ (CIS), tumour-size, reviewed grade and reviewed stage. RESULTS: With a mean follow-up of 6.4 years, there was disease progression in 48 (29%) patients and 26 (16%) died from BC. Associated CIS was found in 55 (34%) patients. After reviewing the slides, 24 (15%) tumours were downstaged to pTa, 134 (82%) remained pT1 and six (4%) were upstaged to > or =pT2. The grade review resulted in 74 G2, 90 G3, 37 low-grade and 127 high-grade lesions for the two systems used. In multivariate analyses, reviewed stage (both P < 0.001) and CIS (P = 0.017 and 0.023) had independent significance for progression and disease-specific survival, respectively. CONCLUSION: A stage review is indicated in pT1 BC, as almost 20% of pT1 tumours were up- or downstaged, and the reviewed stage predicted the patient's prognosis. Hence, pathology review identified patients with different prognoses who might benefit from other treatment strategies than BCG. We confirmed that CIS is an unfavourable sign in pT1 bladder cancer. PMID- 20002440 TI - Chronic cortical and subcortical pathology with associated neurological deficits ensuing experimental herpes encephalitis. AB - Long-term neurological sequela is common among herpes simplex encephalitis (HSE) survivors. Animal models for HSE are used to investigate mechanisms of acute disease, but little has been done to model chronic manifestations of HSE. The current study presents a detailed, systematic analysis of chronic neuropathology, including characterization of topography and sequential progression of degenerative lesions and inflammation. Subsequent to intranasal HSV-1 infection, inflammatory responses that were temporally and spatially distinct persisted in infected cortical and brain stem regions. Neutrophils were present exclusively within the olfactory bulb and brain stem regions during the acute phase of infection, while the chronic inflammation was marked by plasma cells, lymphocytes and activated microglia. The chronic lymphocytic infiltrate, cytokine production, and activated microglia were associated with the loss of cortical neuropile in the entorhinal cortex and hippocampus. Animals surviving the acute infection showed a spectrum of chronic lesions from decreased brain volume, neuronal loss, activated astrocytes, and glial scar formation to severe atrophy and cavitations of the cortex. These lesions were also associated with severe spatial memory deficits in surviving animals. Taken together, this model can be utilized to further investigate the mechanisms of neurological defects that follow in the wake of HSE. PMID- 20002441 TI - Mantle cell lymphoma shows three morphological evolutions of classical, intermediate, and aggressive forms, which occur in parallel with increased labeling index of cyclin D1 and Ki-67. AB - Although the 2008 World Health Organization classification defines two subtypes of mantle cell lymphoma (MCL), classical and aggressive, we often encounter MCL with both features in the same site. We named this feature "MCL with focal aggressive form (intermediate MCL)". In the present study, we reclassified 237 patients with cyclin D1 (CCND1)-positive MCL on the basis of the concept of intermediate MCL, and analyzed the correlation of this reclassification with immunohistochemical detection of CCND1, Ki-67, p53, p27(Kip1), and p21(WAF/Cip1). The median overall survival was 77, 31, and 18 months for classical, intermediate, and aggressive MCL, respectively, showing a statistically significant difference (P < 0.0001). The expression levels of CCND1, Ki-67, p53, and p21(WAF/Cip1) in aggressive MCL (mean 80.1 +/- 27.8%, 73.7 +/- 28.9%, 31.0 +/ 69.0%, and 10.4 +/- 24.8%, respectively) were higher than those in classical MCL (mean 58.1 +/- 36.7%, 25.2 +/- 25.5%, 6.5 +/- 24.3%, and 2.5 +/- 13.0%, respectively) and intermediate MCL (mean 75.7 +/- 31.4%, 30.8 +/- 33.3%, 21.0 +/- 57.4%, and 4.8 +/- 16.5%, respectively). Significantly different levels of Ki-67 and p21(WAF/Cip1) were only recognized between intermediate and aggressive (P < 0.05 and P < 0.0001, respectively), whereas those of CCND1 and p53 were only between classical and intermediate (P < 0.0001 and P < 0.05, respectively). There were no significant differences in p27(Kip1) among the three groups. The subsequent discriminant analysis with independent prognostic factors clearly demonstrated that the morphological evolution of MCL occurs in parallel with increased labeling index of CCND1 and Ki-67. The diagnosis of intermediate MCL thus proved to be of major significance and should enable the design of more tailored therapies. PMID- 20002442 TI - Abstracts from the First Excellence in Paediatrics Conference. December 3-6,2009. Florence, Italy. PMID- 20002446 TI - Epicutaneous immunotherapy on intact skin using a new delivery system in a murine model of allergy. AB - BACKGROUND: Allergen-specific immunotherapy, subcutaneous immunotherapy (SCIT) or oral, has been used for almost a century to redirect inappropriate immune responses in atopic patients. A new mode of administration through the intact skin [epicutaneous immunotherapy (EPIT)], using an original epicutaneous delivery system, may represent an alternative to these classical methods. OBJECTIVE: Proof of concept of efficacy of EPIT on intact skin in mice sensitized to aeroallergens or food allergens. METHODS: Mice were sensitized to pollen (n=18), house dust mite (HDM, n=24), ovalbumin (OVA, n=18) or peanut (n=18), and allocated to four groups: EPIT, SCIT, not treated (NT) and control. Specific Ig (sIg)E, sIgG1 and sIgG2a were monitored. After 8 weeks of treatment, plethysmography was performed after aerosol provocation with appropriate allergens. RESULTS: At the highest doses of methacholine, pause enhancement (Penh) values were significantly decreased in the EPIT group vs. the sensitized NT groups (7.5 vs. 12.3 - pollen, 7.6 vs. 8.9 - HDM, 11.5 vs. 14.5 - OVA, 7.6 vs. 12.8 - peanut, respectively) (P<0.05). With all the allergens tested, Penh values were similar in SCIT, EPIT and control. IgG2a for pollen, HDM, OVA and peanuts were significantly increased in the EPIT group vs. NT: 0.97 vs. 0.42 microg/mL, 2.5 vs. 0.46 microg/mL, 0.39 vs. 0.05 microg/mL and 15.0 vs. 5.5 microg/mL, respectively (P<0.05). There were no significant differences between EPIT and SCIT groups. The IgE/IgG2a ratio decreased significantly in the EPIT group for the four allergens from 70 to 58 (pollen), 175 to 26 (HDM), 5433 to 120 (OVA) and 49 to 6 (peanut), respectively (P<0.05). CONCLUSION: In mice sensitized to the four allergens tested, EPIT was as efficacious as SCIT, considered as the reference immunotherapy. These first results have to be confirmed by clinical studies. PMID- 20002448 TI - Flk-1+ mesenchymal stem cells aggravate collagen-induced arthritis by up regulating interleukin-6. AB - The immunomodulatory ability of mesenchymal stem cells (MSCs) may be used to develop therapies for autoimmune diseases. Flk-1(+) MSCs are a population of MSCs with defined phenotype and their safety has been evaluated in Phase 1 clinical trials. We designed this study to evaluate whether Flk-1(+) MSCs conferred a therapeutic effect on collagen-induced arthritis (CIA), an animal model of rheumatic arthritis, and to explore the underlying mechanisms. Flk-1(+) MSCs, 1-2 x 10(6), were injected into CIA mice on either day 0 or day 21. The clinical course of arthritis was monitored. Serum cytokine profile was determined by cytometric bead array kit or enzyme-linked immunosorbent assay. Flk-1(+) MSCs and splenocytes co-culture was conducted to explore the underlying mechanisms. Flk 1(+) MSCs did not confer therapeutic benefits. Clinical symptom scores and histological evaluation suggested aggravation of arthritis in mice treated with MSCs at day 21. Serum cytokine profile analysis showed marked interleukin (IL)-6 secretion immediately after MSC administration. Results of in vitro culture of splenocytes confirmed that the addition of Flk-1(+) MSCs promoted splenocyte proliferation and increased IL-6 and IL-17 secretion. Moreover, splenocyte proliferation was also enhanced in mice treated with MSCs at day 21. Accordingly, MSCs at low concentrations were found to promote lipopolysaccharide-primed splenocytes proliferation in an in vitro co-culture system. We propose that Flk 1(+) MSCs aggravate arthritis in CIA model by at least up-regulating secretion of IL-6, which favours Th17 differentiation. When Flk-1(+) MSCs are used for patients, we should be cautious about subjects with rheumatoid arthritis. PMID- 20002447 TI - Systemic and local anti-C5 therapy reduces the disease severity in experimental autoimmune uveoretinitis. AB - Activation of complement occurs during autoimmune retinal and intraocular inflammatory disease as well as neuroretinal degenerative disorders. The cleavage of C5 into fragments C5a and C5b is a critical event during the complement cascade. C5a is a potent proinflammatory anaphylatoxin capable of inducing cell migration, adhesion and cytokine release, while membrane attack complex C5b-9 causes cell lysis. Therapeutic approaches to prevent complement-induced inflammation include the use of blocking monoclonal antibodies (mAb) to prevent C5 cleavage. In these current experiments, the rat anti-mouse C5 mAb (BB5.1) was utilized to investigate the effects of inhibition of C5 cleavage on disease progression and severity in experimental autoimmune uveoretinitis (EAU), a model of organ-specific autoimmunity in the eye characterized by structural retinal damage mediated by infiltrating macrophages. Systemic treatment with BB5.1 results in significantly reduced disease scores compared with control groups, while local administration results in an earlier resolution of disease. In vitro, contemporaneous C5a and interferon-gamma signalling enhanced nitric oxide production, accompanied by down-regulation of the inhibitory myeloid CD200 receptor, contributing to cell activation. These experiments demonstrate that C5 cleavage contributes to the full expression of EAU, and that selective C5 blockade via systemic and local routes of administration can suppress disease. This presents great therapeutic potential to protect against tissue damage during autoimmune responses in the retina or inflammation-induced degenerative disease. PMID- 20002450 TI - Respiratory disease in Niemann-Pick type C2 is caused by pulmonary alveolar proteinosis. AB - Niemann-Pick diseases are hereditary neurovisceral lysosomal lipid storage disorders, of which the rare type C2 almost uniformly presents with respiratory distress in early infancy. In the patient presented here, the NPC2 exon 4 frameshift mutation c.408_409delAA caused reduced NPC2 protein levels in serum and lung lavage fluid and the synthesis of an aberrant, larger sized protein of around 28 kDa. Protein expression was strongly reduced also in alveolar macrophages. The infant developed failure to thrive and tachypnea. Lung lavage, computer tomography, and histology showed typical signs of pulmonary alveolar proteinosis with an abnormal intraalveolar accumulation of surfactant as well as macrophages. An NPC2-hypomorph animal model also showed pulmonary alveolar proteinosis and accumulation of macrophages in the lung, liver, and spleen long before the mice died. Due to the elevation of cholesterol, the surfactant had an abnormal composition and function. Despite the removal of large amounts of surfactant from the lungs by therapeutic lung lavages, this treatment was only temporarily successful and the infant died of respiratory failure. Our data indicate that respiratory distress in NPC2 disease is associated with a loss of normal NPC2 protein expression in alveolar macrophages and the accumulation of functionally inactive surfactant rich in cholesterol. PMID- 20002451 TI - Not identical: twins studies to reveal epigenetic differences. PMID- 20002449 TI - Administration of a probiotic associated with nasal vaccination with inactivated Lactococcus lactis-PppA induces effective protection against pneumoccocal infection in young mice. AB - Streptococcus pneumoniae is a serious public health problem, especially in developing countries, where available vaccines are not part of the vaccination calendar. We evaluated different respiratory mucosa immunization protocols that included the nasal administration of Lactococcus lactis-pneumococcal protective protein A (PppA) live, inactivated, and in association with a probiotic (Lc) to young mice. The animals that received Lc by the oral and nasal route presented the highest levels of immunoglobulin (Ig)A and IgG anti-PppA antibodies in bronchoalveolar lavages (BAL) and IgG in serum, which no doubt contributed to the protection against infection. However, only the groups that received the live and inactivated vaccine associated with the oral administration of the probiotic were able to prevent lung colonization by S. pneumoniae serotypes 3 and 14 in a respiratory infection model. This would be related to a preferential stimulation of the T helper type 1 (Th1) cells at local and systemic levels and with a moderate Th2 and Th17 response, shown by the cytokine profile induced in BAL and by the results of the IgG1/IgG2a ratio at local and systemic levels. Nasal immunization with the inactivated recombinant strain associated with oral Lc administration was able to stimulate the specific cellular and humoral immune response and afford protection against the challenge with the two S. pneumoniae serotypes. The results obtained show the probiotic-inactivated vaccine association as a valuable alternative for application to human health, especially in at-risk populations, and are the first report of a safe and effective immunization strategy using an inactivated recombinant strain. PMID- 20002452 TI - A novel nonsense mutation in CUL4B gene in three brothers with X-linked mental retardation syndrome. AB - Cabezas syndrome (MIM 300354) is a recently identified syndromic form of X-linked mental retardation (XLMR) caused by mutations in the CUL4B gene. In total, nine XLMR families carrying mutations in the CUL4B gene have been described to date. Here, we present a detailed clinical phenotype of three affected brothers of Polish descent. Based on the symptoms, we made a clinical diagnosis of Cabezas syndrome, which was subsequently confirmed by identification of a novel nonsense mutation (c.2107A-->T, p.703K-->X) in exon 18 of the CUL4B gene. The mutation was inherited from an asymptomatic mother and was present in all three affected brothers. The patients presented with typical features of Cabezas syndrome, such as severe mental retardation, speech impairment, hyperactivity, seizures, intention tremor, inguinal hernia, small feet, and craniofacial dysmorphism. In addition to previously described symptoms, syndactyly of the second and third toes and skin manifestations (hyperhydrosis and keratosis pilaris) were present in our cases. Our report provides further support that Cabezas syndrome is a recognizable syndromic form of XLMR. We conclude that the CUL4B gene should be screened in males with severe speech impairment and primary intention tremor, especially if characteristic facial dysmorphism is also present. PMID- 20002453 TI - Does family history predict the age at onset of new breast cancers in BRCA1 and BRCA2 mutation-positive families? AB - Women who carry BRCA mutations are advised to begin breast cancer screening based on the age-specific risks of breast cancer development. It is not clear to what extent the family history of breast cancer influences age of onset. We evaluated the use of family history to predict the age of breast cancer onset in BRCA mutation carriers. Pedigrees from an Ontario-based registry were reviewed to identify the index case of breast cancer (most recent diagnosis) and other family cases of breast cancer. The youngest age of breast cancer diagnosis and mean age at breast cancer diagnosis in the other family cases were compared to the age of onset in the index case. The 260 BRCA1 and 213 BRCA2 pedigrees were reviewed. In BRCA2 families, the index case was diagnosed on average at 44.4 years when the youngest reported family case was less than or equal to 35 years, compared to 51.9 years when the earliest cases were diagnosed after age 50 (p = 0.04). A modest trend was seen for BRCA1 carriers, but this was not statistically significant. To a small extent, the onset of breast cancer in a BRCA2 mutation carrier can be predicted from her family history of cancer, however, the trend is modest and should not alter clinical recommendations regarding initiation of screening. PMID- 20002455 TI - DISC1 duplication in two brothers with autism and mild mental retardation. AB - We describe the identification and delineation of an inherited 2.07 Mb microduplication in 1q42.2 in two brothers with autism and mild mental retardation. Since this duplication was not present in 1577 Belgian persons, we consider this as an extremely rare variant which has the potential to provide further insight into the genetics of autism. The duplication contains seven genes including the DISC1 gene, an interesting candidate gene that has been associated to schizophrenia, bipolar disorder, autism and Asperger syndrome. In this report we describe additional analyses undertaken to investigate the causal relationship of the duplication to the autism phenotype. We conclude that the 1q42.2 microduplication probably confers susceptibility to autism in the current family. This study is a typical illustration of the difficult interpretation of causality of a very rare variant in neuropsychiatric disease and the challenge of genetic counselling in a particular family. PMID- 20002456 TI - Prenatal diagnosis of Duchenne muscular dystrophy by comparative genomic hybridization. PMID- 20002458 TI - Identities and frequencies of BMPR2 mutations in Chinese patients with idiopathic pulmonary arterial hypertension. PMID- 20002457 TI - Adverse effects of trichothiodystrophy DNA repair and transcription gene disorder on human fetal development. AB - The effects of DNA repair and transcription gene abnormalities in human pre-natal life have never been studied. Trichothiodystrophy (TTD) is a rare (affected frequency of 10(-6)) recessive disorder caused by mutations in genes involved in nucleotide excision repair (NER) pathway and in transcription. Based on our novel clinical observations, we conducted a genetic epidemiologic study to investigate gestational outcomes associated with TTD. We compared pregnancies resulting in TTD-affected offspring (n = 24) with respect to abnormalities during their antenatal and neonatal periods to pregnancies resulting in their unaffected siblings (n = 18), accounting for correlation, and to population reference values. Significantly higher incidence of several severe gestational complications was noted in TTD-affected pregnancies. Small for gestational age (SGA) <10th percentile [Relative risk (RR ) = 9.3, 95% CI = 1.4-60.5, p = 0.02], SGA <3rd percentile (RR = 7.2, 95% CI = 1.1-48.1, p = 0.04), and neonatal intensive care unit (NICU) hospitalization (RR = 6.4, 95% CI = 1.4-29.5, p = 0.02) occurred more frequently among TTD-affected neonates compared with their unaffected siblings. Compared with reference values from general obstetrical population, pregnancies that resulted in TTD-affected infants were significantly more likely to be complicated by hemolysis, elevated liver enzymes and low platelets (HELLP) syndrome (RR = 35.7, 95% CI = 7.6-92.5, p = 0.0002), elevated mid-trimester maternal serum human chorionic gonadotropin (hCG) levels (RR = 14.3, 95% CI = 7.0-16.6, p < 0.0001), SGA <3rd percentile (RR = 13.9, 95% CI = 7.4-21.1, p < 0.0001), pre-term delivery (<32 weeks) (RR = 12.0, 95% CI = 4.9 21.6, p < 0.0001), pre-eclampsia (RR = 4.0, 95% CI = 1.6-7.4, p = 0.006), and decreased fetal movement (RR = 3.3, 95% CI = 1.6-5.2, p = 0.0018). Abnormal placental development is an underlying mechanism that may explain the constellation of observed complications in our study. Thus, we hypothesize that TTD DNA repair and transcription genes play an important role in normal human placental development. PMID- 20002459 TI - Gene copy number variation and common human disease. AB - Variation in gene copy number is increasingly recognized as a common, heritable source of inter-individual differences in genomic sequence. The role of copy number variation is well established in the pathogenesis of rare genomic disorders. More recently, germline and somatic copy number variation have been shown to be important pathogenic factors in a range of common diseases, including infectious, autoimmune and neuropsychiatric diseases and cancer. In this review, we describe the range of methods available for measuring copy number variants (CNVs) in individuals and populations, including the limitations of presently available assays, and highlight some key examples of common diseases in which CNVs have been shown clearly to have a pathogenic role. Although there has been major progress in this field in the last 5 years, understanding the full contribution of CNVs to the genetic basis of common diseases will require further studies, with more accurate CNV assays and larger cohorts than have presently been completed. PMID- 20002460 TI - Experts' opinions on ethical issues of genetic research into Alzheimer's disease: results of a Delphi study in the Netherlands. AB - Most publications on the ethical aspects of genetic research into Alzheimer's Disease (AD) concentrate on the differences between the opinions of professionals and non-professionals. Differences in rating of morally relevant issues between groups of professionals have not yet been described. A modified Delphi study in two rounds was held to identify differences between groups of experts (i.e. clinicians, representatives of patient organisations, ethicists and persons with a commercial background). The strongest correlation was found between the opinions of ethicists and representatives of patient organisations (0.67) and between clinicians and ethicists (0.62). Moderate correlation (0.55) was found between the opinions of clinicians and representatives of patient organisations. Persons with a commercial background showed a weak correlation with clinicians (0.41), ethicists (0.35) and representatives of patient organisations (0.30). These differences in rating of morally relevant issues between various professional groups are relevant for clinical practice and dementia care, particularly the different rating of prenatal diagnosis found between clinicians and representatives of patient organisations. Interdisciplinary consultations between various professional groups -including at least researchers, clinicians and ethicists -are recommended to guarantee that all considerations will be incorporated into the debate on ethical issues of genetic research into AD. PMID- 20002461 TI - Mutational study in the PDHA1 gene of 40 patients suspected of pyruvate dehydrogenase complex deficiency. AB - We screened for PDHA1 mutations in 40 patients with biochemically demonstrated PDHc deficiency or strong clinical suspicion, and found changes with probable pathological significance in 20. Five patients presented new mutations: p.A169V, c.932_938del, c.1143_1144 ins24, c.1146_1159dup and c.510-30G> A, this latter is a new undescribed cause of exon 6 skipping. Another four mutations have been found, and previously reported, in our patients: p.H113D, p.P172L, p.Y243del and p.Y369Q. Eleven patients presented seven known mutations: p.R127Q, p.I166I, p.A198T, p.R263G, p.R302C, p.R378C and c.1142_1145dup. The latter three were found in more than one unrelated patient: p.R302C was detected in a heterozygous girl and a mosaic male, p.R378C in two males and finally, c.1142_1145dup in three females; only one in 20 mothers was found to be a carrier (p.R263G). Apart from those 20 patients, the only alteration detected in one girl with clear PDHc and PDH-E1 deficiency was the silent change c.396A> C (p.R132R), and other eight PDHc deficient patients carry combinations of known infrequent polymorphisms that are overrepresented among our 20 unsolved patients. The importance of these changes on PDH activity is unclear. Investigations in the other PDHc genes are in course in order to elucidate the genetic defect in the unresolved patients. PMID- 20002462 TI - The effect of titanium topography features on mesenchymal human stromal cells' adhesion. AB - OBJECTIVE: The aim of this study was to evaluate the effect of two kinds of dental implants surfaces with their own characteristics on human marrow stromal cells' adhesion. MATERIAL AND METHODS: Fifty-six titanium discs (28 machined and 28 acid etched) were used. Machined (MS) and acid-etched surfaces (ES) were analyzed using a scanning electron microscope, energy dispersing spectroscopy (EDS), contact angle analysis and human marrow stromal cells' culture. RESULTS: Significant differences were observed in the topography and wetability of the tested surfaces. However, etched surfaces presented a high level of wetability when compared with machined surfaces. Contact angles showed considerable differences between etched and machined surfaces (Friedman test P<0.05). EDS analysis showed the same composition on both the surfaces tested. Counting of adhered cells on both types of surfaces showed that there is no statistical significance in human marrow stromal cells' adhesion after 18 h (Mann-Whitney test P>0.05). CONCLUSION: The present study concludes that modifications on the titanium implant surfaces roughness may promote differences in the morphology of bone marrow stromal cells. Nevertheless, in this microenvironment, no interference in the adhesion phenomenon was noted. PMID- 20002463 TI - Indications for combined liver and kidney transplantation: propositions after a 23-yr experience. AB - The frequency of combined liver and kidney transplants (CLKT) persists despite the pronounced scarcity of organs. In this review, we sought to ascertain any factors that would reduce the use of these limited commodities. Seventy-five adult CLKT were performed over a 23-yr period at our center, 29 (39%) of which occurred during the Model for End-stage Liver Disease (MELD) era. Overall, patient survival rates were 82%, 73%, and 62% at one, three, and five yr, respectively. There was no difference in patient survival based either on pre transplant hemodialysis status or by glomerular filtration rate (GFR) at the time of transplant. Patients undergoing a second CLKT or a liver retransplantation at the time of CLKT had a survival rate of 30% at three months. In the MELD era, patient survival was unchanged (p = NS) despite an older recipient population (p = 0.0029) and a greater number of hepatitis C patients (p = 0.0428). In summary, patients requiring liver retransplantation with concomitant renal failure should be denied CLKT. Renal allografts may also be spared by implementing strict criteria for renal organ allocation (GFR < 30 mL/min at the time of evaluation) and considering the elimination of preemptive kidney transplantation in CLKT. PMID- 20002464 TI - Transplantation tourism: high risk for the recipients. AB - BACKGROUND: The shortage of donor organ supply is forcing patients with end-stage renal disease to alternative searches. The aim of this study is to present the clinical and laboratory data of five patients who were transplanted in Egypt from paid living-unrelated donors and followed at our institution. METHODS: Five patients (four male, one female, mean age 51 yr) were included in this retrospective study. RESULTS: All allografts still have good function with a mean serum creatinine level of 0.9 mg/dL. Surgical and medical problems were common such as wound infection (n = 3), evisceration (n = 2), deep vein thrombosis (n = 2), unexplained abdominal incision requiring removal of an abdominal surgical compress left in situ during previous surgery, placement of allograft on the side of an unrepaired indirect inguinal hernia and transplant pyelonephritis. CONCLUSION: Although recent developments increased success in renal transplantation, receiving a kidney from a paid living donor at a commercial transplant center still carries great risks for the recipient. PMID- 20002465 TI - Hand-assisted versus total laparoscopic live donor nephrectomy: comparison and technique evolution at a single center in Taiwan. AB - PURPOSE: To compare the outcome of hand-assisted laparoscopic live donor nephrectomy (HLDN) and total laparoscopic live donor nephrectomy (TLDN) in a single center. METHODS: The demographics, complications, and outcomes were compared between successfully performed 51 HLDN and 42 TLDN. RESULTS: The patients' demographics including body mass index were all similar. Four conversions were excluded for the outcome analysis. The operation time of HLDN group (188 +/- 62 min) was shorter, although not significantly, than that of TLDN group's (207 +/- 30 min) (p = 0.065). However, the operation time of the first 24 cases (237 +/- 66 min) was significantly longer than that of the later 69 performed (180 +/- 35 min). The warm ischemia time was shorter in HLDN (2.5 +/- 1.3 min) compared to that of TLDN (4.1 +/- 1.7 min) (p < 0.01), but the serum creatinine values (mg/dL) of recipients were equivalent (HLDN/TLDN = 1.18 +/- 0.3:1.14 +/- 0.3, p = 0.587). There was no difference in the length of hospital stay (6.7 vs. 6.4 d, p = 0.475). There was no graft loss, but one ureter stricture (HLDN group) and one urinary leakage (TLDN group) were recorded. CONCLUSIONS: Both HLDN and TLDN are effective and safe as reflected in graft functions and limited complications. There was a learning curve in establishing the technique of laparoscopic donor nephrectomy. PMID- 20002466 TI - The impact of sirolimus on hepatocyte proliferation after living donor liver transplantation. AB - BACKGROUND: There is a lack of data on the use of sirolimus after partial liver transplantation, especially regarding its impact on post-transplant regeneration. METHODS: We reviewed adult living donor transplantations, with de novo sirolimus (n = 7) and without sirolimus (n = 21). Liver biopsies were stained for KI-67, a proliferation marker. Controls included specimens with normal liver parenchyma (n = 13). RESULTS: Both groups had similar demographics, graft and patient survival and complication rates. During the first six wk and over the whole first year post-transplant, the use of sirolimus was associated with lower levels of hepatocyte proliferation compared to sirolimus-free patients, (overall, 0.3 [0 7.2] vs. 3 [0-49] KI-67 positive hepatocytes per high power field, p <= 0.05). The levels observed in the sirolimus group were similar to those seen in non transplanted control patients with normal parenchyma (0.2 [0-1.3], p = NS). Post transplant hepatocyte proliferation correlated with the serum levels of sirolimus (p <= 0.05), but not with those of tacrolimus or with the dose of mycophenolate mofetil (p = 0.9 and 0.3, respectively). CONCLUSIONS: These data suggest that sirolimus is associated with decreased post-transplant hepatocyte proliferation. The clinical significance of this observation remains to be fully determined. PMID- 20002467 TI - Commentary: Rectal cancer patients are often not suitable for laparoscopic surgery. PMID- 20002468 TI - Role of glucotoxicity and lipotoxicity in the pathophysiology of Type 2 diabetes mellitus and emerging treatment strategies. AB - Type 2 diabetes mellitus is a disease characterized by persistent and progressive deterioration of glucose tolerance. Both insulin resistance and impaired insulin secretion contribute to development of Type 2 diabetes. However, whilst insulin resistance is fully apparent in the pre-diabetic condition, impairment of insulin secretion worsens over the time, being paralleled by a progressive decline in both pancreatic B-cell function and B-cell mass. Intense research has identified a number of genetic variants that may predispose to impaired B-cell function, but such predisposition can be precipitated and worsened by toxic effects of hyperglycaemia (glucotoxicity) and elevated levels of free fatty acids (lipotoxicity). All these aspects of the pathogenesis of Type 2 diabetes are discussed in this review. Moreover, treatments that target reduction in glucotoxicity or lipotoxicity are outlined, including emerging strategies that target the role of glucagon-like peptide 1 and sodium glucose co-transporter 2. PMID- 20002469 TI - Perinatal risk factors for early onset of Type 1 diabetes in a 2000-2005 birth cohort. AB - AIMS: To examine perinatal risk factors for the onset of Type 1 diabetes before 6 years of age, in a 2000-2005 Australian birth cohort. METHODS: Data from longitudinally linked delivery and hospital admission records (until June 2007) were analysed. Diabetes in mothers and children was identified from International Classification of Diseases 10 diagnosis codes in the hospital records. RESULTS: There were 272 children admitted to hospital with a first diagnosis of diabetes out of 502 040 live births. Incidence for the infants born in 2000 was 16.0 per 100 000 person-years. Maternal Type 1 diabetes was a significant risk factor [crude relative risk (RR) 6.33], but maternal Type 2 diabetes and gestational diabetes were not significantly associated with diabetes in the child. Late preterm birth (34-36 weeks) (RR 1.64) and caesarean section (RR 1.30) increased the risk of a diabetes admission. Size-for-gestational-age was significantly associated with onset of diabetes (small-for-gestational age RR 0.48), but neither birth weight categories nor birth weight as a continuous variable were associated with risk of diabetes. Increasing maternal age was associated with an increased risk of diabetes in the child (RR 1.13 for each additional 5 years of age). CONCLUSIONS: This study identified risk factors associated with onset of Type 1 diabetes before 6 years of age, in a recent birth cohort. Size-for gestational-age had a consistent association with risk of early onset of Type 1 diabetes, small size being protective. Size-for-gestational-age measures should be preferred to birth weight thresholds when assessing risk of diabetes. PMID- 20002470 TI - Evaluation of proposed oral disposition index measures in relation to the actual disposition index. AB - Aims While the disposition index provides a useful measure of B-cell function, its calculation requires the performance of a frequently sampled intravenous glucose tolerance test (FSIVGTT). Recently, the demonstration of a hyperbolic relationship between indices of insulin secretion and insulin sensitivity derived from the oral glucose tolerance test (OGTT) has led to the introduction of two novel OGTT-based measures of B-cell function analogous to the disposition index: (i) the insulin secretion-sensitivity index-2 (ISSI-2) (defined as the ratio of the area-under-the-insulin-curve to the area-under-the-glucose curve, multiplied by the Matsuda index) and (ii) insulinogenic index (IGI)/fasting insulin. However, neither of these two measures has been directly compared with the disposition index. Methods Two hundred and thirteen non-diabetic children (122 boys, 91 girls) underwent both OGTT and FSIVGTT, allowing for the calculation of ISSI-2, IGI/fasting insulin and the disposition index. Results ISS1-2 and IGI/fasting insulin were strongly correlated with each other (r = 0.82, P < 0.0001). Both measures correlated with the disposition index, with ISSI-2 showing a modestly stronger association (ISSI-2: r = 0.24, P = 0.0003; IGI/fasting insulin: r = 0.21, P = 0.0022). Standardized linear regression analyses confirmed that the relationship between log ISSI-2 and the disposition index (standardized regression coefficient = 0.224, P = 0.001) was stronger than that between log IGI/fasting insulin and the disposition index (standardized regression coefficient = 0.166, P = 0.015). Conclusions The OGTT-derived measures ISSI-2 and IGI/fasting insulin exhibit modest correlations with the disposition index. These relationships require further assessment in other patient populations. PMID- 20002471 TI - Metabolic syndrome and alanine aminotransferase: a global perspective from the NAVIGATOR screening population. AB - AIMS: Non-alcoholic fatty liver disease (NAFLD) is associated with features of the metabolic syndrome (MetS) and may be an expression of the syndrome within the liver. Using screening data from the Nateglinide And Valsartan in Impaired Glucose Tolerance Outcomes Research (NAVIGATOR) study (n = 42 149), we examined whether alanine aminotransferase (ALT), a biomarker for NAFLD, clustered with features of MetS and whether the clusters differed across global geographic regions. METHODS: Exploratory factor analysis using principle components analysis was applied to data drawn from the NAVIGATOR screening population (n = 41 111). Demographic data, anthropomorphic measurements and blood pressure (BP) collected during the screening visit, as well as blood samples analysed for ALT, total cholesterol, triglycerides, high-density lipoprotein, low-density lipoprotein, and fasting and 2-h glucose measures after an oral glucose tolerance test were used for our analysis. RESULTS: Two factors, interpreted as lipid (Factor 1), and BP/obesity (Factor 2) were identified, explaining approximately 50% of the variance in the overall population. Similar patterns of aggregation were reproducible across all geographic regions except Asia, where fasting glucose loaded more consistently on Factor 1. ALT loaded with mean arterial pressure, fasting glucose and waist circumference except in Asia, where it loaded only with mean arterial pressure and waist circumference. CONCLUSIONS: ALT aggregated with components of MetS, and the pattern of aggregation of ALT with other features of MetS was similar across regions except Asia, possibly indicating a different pathophysiology for NAFLD in Asia. Predictive models of NAFLD may need to be adjusted for regional and ethnic differences. PMID- 20002472 TI - Incidence of Type 2 diabetes in the elderly German population and the effect of clinical and lifestyle risk factors: KORA S4/F4 cohort study. AB - AIMS: To determine the incidence of Type 2 diabetes in an elderly population in Germany and its association with clinical and lifestyle factors. METHODS: Oral glucose tolerance tests (OGTT, World Health Organization criteria) were carried out in a random sample of 1353 subjects (age group 55-74 years; 62% response) in Augsburg (Southern Germany) (1999-2001). The cohort was re-investigated in 2006 2008. Of those individuals without diabetes (baseline), 887 (74%) participated in the follow-up. RESULTS: Ninety-three (10.5%) developed diabetes during the 7-year follow-up period {standardized incidence rates [95% confidence interval (CI)] per 1000 person-years: total 15.5; 12.6, 19.1; men 20.2; 15.6, 26.1; women 11.3; 7.9, 16.1}. In both sexes, those who developed diabetes were slightly older, were more obese, had a more adverse metabolic profile (higher glucose values, HbA(1c), fasting insulin, uric acid, and triglycerides) and were more likely to have hypertension at baseline than were participants remaining free of diabetes (P < 0.05). On stepwise logistic regression, age, parental diabetes, body mass index, uric acid, current smoking, HbA(1c) and fasting and 2-h glucose (OGTT) were strong predictors of diabetes incidence. The risk of diabetes was higher in subjects with isolated impaired glucose tolerance (odds ratio 8.8; 95% CI 5.0, 15.6) than in isolated impaired fasting glucose (4.7; 2.2, 10.0), although the difference did not reach statistical significance. CONCLUSIONS: For the first time, we have estimated the incidence of Type 2 diabetes in an elderly German cohort and demonstrated that it is among the highest in Europe. The OGTT appears to be useful in identifying individuals with high Type 2 diabetes risk. Our results support a role of smoking in the progression to diabetes. PMID- 20002473 TI - Increasing trend in the prevalence of Type 2 diabetes and pre-diabetes in the Chinese rural and urban population in Qingdao, China. AB - AIMS: To determine the secular trend of prevalence of Type 2 diabetes and pre diabetes in a Chinese population from 2001 to 2006. METHODS: Two consecutive population-based surveys for diabetes were conducted in a randomly selected population aged 35-74 years and living in Qingdao, China in 2001-2002 (n = 10854) and 2006 (n = 4416). All participants underwent standardized 2-h 75-g oral glucose tolerance tests (OGTTs), along with fasting capillary plasma glucose (FCG) tests in 2006. One urban community underwent OGTTs directly in 2002 (n = 1815), while a two-step screening strategy using FCG as a first-line screening test followed by OGTTs was used in 9039 individuals in 2001. Diabetes and pre diabetes was defined according to the 2006 World Health Organization/International Diabetes Federation criteria. RESULTS: Based on the results of direct OGTTs, the age-standardized prevalence of diabetes and pre diabetes in urban areas was 12.2 and 15.4% in 2002, whereas the prevalences were 18.8 and 28.7% in urban areas and 14.1 and 20.2% in rural areas in 2006 (P < 0.001, in urban areas). Using the two-step screening strategy, the prevalence of diabetes in 2001 was 10.1% in urban and 7.7% in rural areas and 13.8% in urban and 12.2% in rural areas in 2006 (P < 0.001). Based on the data of the 2006 survey, the two-step screening strategy missed 30.2% of diabetes cases when compared with the number defined by the direct OGTT approach. CONCLUSIONS: Qingdao has experienced a marked increase in the prevalence of diabetes and pre diabetes in the past 5 years. Intervention to prevent a further increase in the prevalence of diabetes is urgently required. PMID- 20002474 TI - Nerve growth factor and expression of its receptors in patients with diabetic neuropathy. AB - AIMS: Low serum nerve growth factor (NGF) levels have been reported in patients with diabetic peripheral neuropathy (DPN), but the role of NGF in the development of neuropathy is unclear. Thus, we investigated the associations of serum NGF level and NGF receptor activity with the presence and severity of DPN. METHODS: One hundred and thirty-six patients with Type 2 diabetes were included in this cross-sectional study. Serum NGF levels were measured by ELISA. Expressions of NGF receptors (TrkA and p75(NTR)) were measured by immunohistochemical staining. The presence and severity of DPN were assessed by neuropathy disability score (NDS) and by corneal nerve fibre length (cNFL) and nerve branch density (cNBD) using in vivo confocal microscopy. RESULTS: Patients with DPN had higher serum NGF levels (56-451 pg/ml) than patients without DPN (4-54 pg/ml). However, in DPN patients, serum NGF was negatively associated with neuropathy severity (mild 222 +/- 64 pg/ml; moderate 114 +/- 17 pg/ml; severe 89 +/- 20 pg/ml). This negative association was consistent in all severity indices (NDS, P < 0.001; cNFL, P < 0.001; cNBD P = 0.010) even after adjustment for age, sex, diabetes duration, insulin use, fasting glucose and glycated haemoglobin. Although NGF receptor activities had significantly (P < 0.05) negative associations with the presence and severity of neuropathy, these associations were not significant when adjusted for other factors. CONCLUSIONS: Serum NGF level was positively associated with the presence of DPN but negatively associated with neuropathy severity in DPN patients. The change in serum NGF might be a consequence of, rather than a contributor to, the early development of DPN. PMID- 20002475 TI - Abnormal liver function tests in patients with Type 1 diabetes mellitus: prevalence, clinical correlations and underlying pathologies. AB - AIMS: To determine the prevalence of elevated alanine transaminase (ALT) in a large cohort of patients with Type 1 diabetes and to examine the clinical correlations and causes. Methods Patients with Type 1 diabetes mellitus were prospectively recruited and ALT, glycated haemoglobin and lipid profile were measured. Patients with Type 2 diabetes mellitus were recruited as a comparison group. PATIENTS: with abnormal ALT were investigated for underlying causes. Prevalence of abnormal ALT was analysed at three separate cut-offs and multivariable analysis used to identify independent risk factors. RESULTS: Nine hundred and eleven with Type 1 diabetes and 963 with Type 2 diabetes were included. The prevalence of elevated ALT was dependent on the cut-off value: > 30 IU/l in males and > 19 IU/l in females, > 50 and > 63 IU/l was 34.5, 4.3 and 1.9%, respectively, in Type 1 diabetes and 51.4, 8.2 and 3.7%, respectively, in Type 2 diabetes. In Type 1 diabetes an elevated ALT was associated with worse glycaemic control, age > 55 years and elevated triglycerides. Investigation of these patients revealed a cause in 43.6% of patients, predominantly non-alcoholic fatty liver disease (NAFLD). CONCLUSIONS: Elevated ALT is not uncommon in Type 1 diabetes and is associated with NAFLD-related risk factors. Patients with Type 1 diabetes and elevated ALT should be investigated as significant abnormalities may be found which are amenable to interventions. PMID- 20002476 TI - Long-term glycaemic effects of pioglitazone compared with placebo as add-on treatment to metformin or sulphonylurea monotherapy in PROactive (PROactive 18). AB - AIMS: To assess the long-term glycaemic effects, concomitant changes in medications and initiation of permanent insulin use (defined as daily insulin use for a period of > or = 90 days or ongoing use at death/final visit) with pioglitazone vs. placebo in diabetic patients receiving metformin or sulphonylurea monotherapy at baseline in the PROspective pioglitAzone Clinical Trial in macroVascular Events (PROactive). METHODS: In PROactive, patients with Type 2 diabetes and macrovascular disease were randomized to pioglitazone (force titrated to 45 mg/day) or placebo, in addition to other existing glucose-lowering therapies. In a post-hoc analysis, we categorized patients not receiving insulin at baseline and treated by oral monotherapy into two main cohorts: add-on to metformin alone (n = 514) and sulphonylurea alone (n = 1001). The follow-up averaged 34.5 months. RESULTS: There were significantly greater reductions in glycated haemoglobin (HbA(1c)) with pioglitazone than with placebo and more pioglitazone-treated patients achieved HbA(1c) targets, irrespective of the baseline oral glucose-lowering regimen and despite a decrease in the use of other glucose-lowering agents. Approximately twice as many in the placebo groups progressed to permanent insulin use than in the pioglitazone groups across the two cohorts: 3.4% for pioglitazone and 6.5% for placebo when added to metformin monotherapy and 6.3% and 14.8%, respectively, when added to sulphonylurea monotherapy. The overall safety of both dual therapies was good. CONCLUSIONS: Intensifying an existing oral monotherapy regimen to a dual oral regimen by adding pioglitazone resulted in sustained improvements in glycaemic control and reduced progression to insulin therapy. The efficacy and safety of adding pioglitazone to either metformin monotherapy or sulphonylurea monotherapy were good. PMID- 20002477 TI - The effect of biopsy-positive silent coeliac disease and treatment with a gluten free diet on growth and glycaemic control in children with Type 1 diabetes. AB - OBJECTIVE: To determine the effect of coeliac disease and treatment with a gluten free diet on growth and glycaemic control in asymptomatic children with Type 1 diabetes. METHODS: Data were compared in children with coeliac disease diagnosed by annual antibody screening and jejunal biopsy and treated with a gluten-free diet (n = 49) against individuals who were antibody negative (n = 49) matched for age, sex and duration of diabetes. RESULTS: No differences in growth were observed. In the years prior to diagnosis of coeliac disease, mean glycated haemoglobin (HbA(1c)) was lower in cases compared with control subjects [8.3 +/- 1.1% vs. 8.7 +/- 0.9%, P = 0.02 (mean +/- sd)]. In cases, HbA(1c) deteriorated 12 months from the start of a gluten-free diet to levels similar to control subjects (8.9 +/- 1.5% vs. 8.8 +/- 1.5%, P-value for analysis of variance = 0.9). In regression analysis, the diagnosis of coeliac disease and start of a gluten-free diet was associated with a rise in HbA(1c) in the first year of treatment [odds ratio 1.56 (95% confidence intervals 1.16-2.10), P = 0.003] after adjusting for insulin dose and regimen and other variables. CONCLUSIONS: In children with Type 1 diabetes, lower HbA(1c) prior to diagnosis of silent coeliac disease rises following treatment with a gluten-free diet to levels similar to those without coeliac disease. Although unproven, these observations may relate to abnormalities at the small bowel mucosa before the appearance of circulating coeliac antibodies. PMID- 20002478 TI - Efficacy and safety of insulin glargine and glimepiride in subjects with Type 2 diabetes before, during and after the period of fasting in Ramadan. AB - AIMS: To determine the safety and efficacy of insulin glargine and glimepiride in patients with Type 2 diabetes before and after Ramadan and during fasting for Ramadan. METHODS: In this open, descriptive, multi-centre, prospective study, insulin-naive (n = 100) or previously insulin-treated (n = 249) patients with Type 2 diabetes received insulin glargine [titrated from 10 U daily according to fasting blood glucose (FBG)] and glimepiride (4 mg daily). The number and type of hypoglycaemic episodes and glycaemic control were assessed before, during and after Ramadan. Bivariate logistic regression analyses were used to identify factors which predicted hypoglycaemia during Ramadan. RESULTS: Only one episode of severe hypoglycaemia occurred in each time period before, during and after Ramadan. Mild hypoglycaemic episodes increased from 156 pre-Ramadan to 346 during Ramadan (P < 0.001) and decreased to 153 post-Ramadan (P = 0.0002).The increase during Ramadan was mainly attributed to increased symptomatic hypoglycaemic episodes. FBG and glycated haemoglobin improved during the titration period and did not change during the rest of the study. Risk of hypoglycaemic events during Ramadan was higher in countries where fasting is strict [odds ratio (OR) 3.69 (2.06-6.63), P < 0.0001]. Lower weight [< 70.0 kg; OR 2.56 (1.46-4.48), P = 0.001] and waist circumference [< 90 cm; OR 3.06 (1.62-5.78), P = 0.001] increased the risk of hypoglycaemia during Ramadan whilst FBG > 6.7 mmol/l [OR 0.3 (0.17-0.54), P < 0.0001] had a protective effect. CONCLUSIONS: Combination of insulin glargine and glimepiride may be used during Ramadan in patients with Type 2 diabetes who wish to fast, provided glimepiride is given at the time of breaking the fast and insulin glargine titrated to provide FBG > 6.7 mmol/l. PMID- 20002479 TI - Performance of HbA(1c) for detecting newly diagnosed diabetes and pre-diabetes in Chinese communities living in Beijing. AB - AIM: To determine the performance of glycated haemoglobin (HbA(1c)) as a screening tool for detecting newly diagnosed diabetes (NDM) and pre-diabetes. METHODS: A diabetes survey was conducted in Beijing among community dwellers who were willing to participate in the survey. Included in the survey were 903 individuals aged 21-79 years without previously diagnosed diabetes and in whom HbA(1c) and other required covariates had been measured. NDM and pre-diabetes (impaired glucose tolerance + impaired fasting glucose) were defined according to the World Health Organization 1999 criteria based on 75-g oral glucose tolerance test. Receiver operating characteristic curve (ROC) was plotted to determine the performance of HbA(1c). RESULTS: The prevalence of NDM and pre-diabetes was 11.1% and 22.4%, respectively. At an optimal HbA(1c) cut-off point of > or = 6.0%, the test gave a sensitivity of 80.0% and a specificity of 89.8% for diagnosing NDM; at an optimal cut-off point of > or = 5.7%, the sensitivity was 59.4% and specificity 73.9% for diagnosing pre-diabetes. Individuals with HbA(1c)> or = 6.0% tended to be more obese than those with HbA(1c) < 6.0%, but blood pressure and lipid profiles did not differ between the two groups. CONCLUSIONS: HbA(1c) as a single screening test is adequate to detect newly diagnosed diabetes but is not able to identify pre-diabetes in this obese Chinese population. PMID- 20002480 TI - Which people with Type 2 diabetes achieve good control of intermediate outcomes? Population database study in a UK region. AB - AIMS: To measure quality of vascular risk factor measurement and control in people with Type 2 diabetes after comprehensive pay-for-performance implementation and to examine variation by patient and practice characteristics. METHODS: Multi-level regression analysis of 10 191 patients with Type 2 diabetes registered with 59 practices in the Tayside region. Quality measures examined were recording of glycated haemoglobin (HbA(1c)), blood pressure (BP), cholesterol and smoking status in the last 12 months; achievement of recommended intermediate outcome targets (HbA(1c)< or = 7.4%, BP < 140/80 mmHg, cholesterol < or = 5.0 mmol/l, not smoking); and simple and all-or-none composite measures. RESULTS: Ninety-five per cent of all recommended processes were received by patients, with 88% of patients receiving all four. Half of all intermediate outcomes targets were achieved, but only 16% of patients achieved all four targets. Process and outcome of care were consistently worse for 1523 (15.0%) patients aged < 55 years. HbA(1c) and BP targets were progressively less likely to be achieved as body mass index increased. Women were less likely to achieve cholesterol targets, but apart from smoking status, there were no associations with socio-economic status. CONCLUSION: Under comprehensive pay-for-performance, process of care is remarkably reliable, but intermediate outcome control less so. Previously identified socio-economic variations in diabetes care have been largely eliminated, but gender inequality is persistent. Younger people were considerably less likely to achieve intermediate outcome targets. Mitigating increased vascular risk in younger patients with Type 2 diabetes presents major challenges for health services in the face of the evolving epidemics of obesity and diabetes. PMID- 20002481 TI - Does an advanced insulin education programme improve outcomes and health service use for people with Type 2 diabetes? A 5-year follow-up of the Newcastle Empowerment course. AB - OBJECTIVE: To show that an advanced diabetes education programme delivers sustained benefits to people with diabetes prescribed insulin and healthcare providers over and above those provided by basic diabetes education. METHODS: An historical cohort study of 68 people with Type 1 and 51 people with Type 2 diabetes on insulin who attended the 4-day Newcastle Empowerment programme in 2001 and 2002 compared with 71 people with Type 1 and 312 people with Type 2 diabetes who attended only the basic 4-day insulin education programme over the same period, followed until 2007. Primary outcome was all hospital admissions and emergency visits; secondary outcomes were the composite of first cardiac event or death and readmission for diabetes complications. Cox-proportional hazards regression was used to analyse Type 1 and Type 2 diabetes separately. RESULTS: The empowerment programme significantly delayed time to first hospital admission/visit for patients with Type 2 diabetes; the hazard ratio (HR) of 0.41 (P = 0.01) translates into a delay of almost 3 years; this was partly driven by a significant reduction in cardiovascular events and mortality (HR = 0.24, P = 0.01). These effects were not seen for people with Type 1 diabetes. CONCLUSIONS: A one-time, advanced diabetes education programme teaching intensive insulin self management with an empowerment style can lead to sustained improvement in patient outcomes and reduce use of hospital services for people with Type 2 diabetes on insulin. PMID- 20002482 TI - Moderating effects of coping styles on anxiety and depressive symptoms caused by psychological stress in Chinese patients with Type 2 diabetes. AB - AIMS: This study aimed to assess possible interactive effects of coping styles and psychological stress on depression and anxiety symptoms in Chinese patients with Type 2 diabetes. METHODS: Three hundred and four patients with Type 2 diabetes underwent a face-to-face interview by trained research staff according to a standardized questionnaire including information on socio-demographic characteristics, psychological stress, coping styles and anxiety and depressive symptoms. The interactive effects of coping styles and psychological stress on depression and anxiety symptoms were assessed by hierarchical multiple regression analyses. RESULTS: There were significant associations of the four domains of psychological stress with anxiety and depressive symptoms, except for the relationship between 'reduced economic condition' and depressive symptoms. 'Negative coping style' significantly increased the level of both anxiety and depressive symptoms; whereas, 'active coping style' and 'avoidant coping style' decreased the risk of depressive symptoms. The interactions of 'negative coping style' with 'worrying about decline in body/physical function' and 'reduced economic condition' significantly increased the risk of anxiety and depressive symptoms, and the interaction of 'social/family crisis caused by the disease' with 'avoidance coping style' and 'worrying about decline in body/physical function' with 'active coping style' significantly decreased the risk of depressive symptoms. CONCLUSIONS: The results of this study suggest that certain coping styles might moderate the association of psychological stress with anxiety and depressive symptoms in Chinese patients with Type 2 diabetes. PMID- 20002483 TI - Handedness, insulin sensitivity and pancreatic B-cell function in Type 2 diabetes. AB - BACKGROUND: Laterality is associated with various health conditions. No study has addressed the influence of handedness on Type 2 diabetes mellitus (T2DM) phenotype, including glucose homeostasis, glucose-lowering therapies and metabolic control. METHODS: Five hundred and seventy-six consecutive adult T2DM outpatients underwent homeostasis model assessment (HOMA) of pancreatic B-cell function (B), insulin sensitivity (S), hyperbolic product (B x S) and age standardized B x S deficit. Right-handed patients (87.5%; RH; n = 504) had similar age, gender, diabetes duration and education than non-right-handed patients (12.5%; non-RH; n = 72). RESULTS: Non-RH were more insulin-sensitive: 66% (39%) vs. 52% (36%) [mean (1 sd); P = 0.0024] and had significantly higher B x S and lower age-adjusted B x S deficit: 35% (20%) vs. 26% (17%) and 1.08% (0.40%) vs. 1.32% (0.55%)/year (non-RH; P = 0.0005 and P < 0.0001, respectively). CONCLUSIONS: Non-right-handed T2DM patients are more insulin-sensitive, have higher hyperbolic product and less age-standardized B x S deficit. These may modulate glucose-lowering therapy requirements and glycaemic control. PMID- 20002484 TI - First trimester serum neutrophil gelatinase-associated lipocalin in gestational diabetes. AB - AIMS: Neutrophil gelatinase-associated lipocalin (NGAL) is highly expressed in damaged epithelia, during inflammation and cardiovascular disease. Soluble NGAL was measured in women who subsequently developed gestational diabetes. METHODS: From a cohort of 908 pregnant outpatients who participated in a screening programme for Down's syndrome at 9-12 weeks of gestation, we considered all 41 women with a singleton pregnancy, who developed gestational diabetes in the previous 12 months, and a control group of 82 normal pregnancies. Circulating NGAL and insulin resistance by homeostasis model assessment ratio (HOMA-IR) were determined in the first trimester. RESULTS: Median serum NGAL concentrations were higher in the gestational diabetes group [51.3 ng/ml (39.8-66.1 ng/ml)] compared with the control group [17.8 ng/ml (15.5-20.9 ng/ml)] (P < 0.001) and positively correlated with HOMA-IR (P < 0.001). CONCLUSIONS: In the first trimester, circulating NGAL was significantly increased in women who subsequently developed gestational diabetes compared with the control group. PMID- 20002485 TI - Short report: suboptimal diabetes care in high-risk diabetic patients attending a specialist retina clinic. AB - AIMS: Individuals with diabetic retinopathy (DR) represent a high-risk group who would benefit from intensive metabolic control and risk factor management. This brief report examines quality of care among diabetic patients attending a tertiary retinal clinic. METHODS: A cross-sectional survey, notes review, and slit-lamp examination was conducted in 139 diabetic patients attending a specialist retinal clinic to assess the quality of comprehensive diabetes care. DR was graded according to the Early Treatment Diabetic Retinopathy Study scale. RESULTS: The prevalence of non-proliferative DR (NPDR) and proliferative DR (PDR) was 39.6 and 35.2%, respectively. The prevalence of microalbuminuria in patients with no DR, NPDR and PDR was 32, 54.1 and 68.8%, respectively. Glycaemic control was suboptimal (mean HbA(1c) 8.0 +/- 1.8%) and 15.8% were current smokers. Drugs affecting the renin-angiotensin system were used by only 61.9% of patients with both DR and microalbuminuria, and aspirin by only 35.3%. CONCLUSIONS: These data suggest that diabetes care in this high-risk population with established microvascular complications was suboptimal. Specialist clinics dealing with diabetic complications may be a setting where quality improvement strategies to reduce morbidity and mortality should be focused. PMID- 20002486 TI - A national survey of the current state of screening services for diabetic retinopathy: ABCD-diabetes UK survey of specialist diabetes services 2006. AB - The main aims were to ascertain the progress made in the implementation of retinal screening services and to explore any barriers or difficulties faced by the programmes. The survey focused on all the essential elements for retinal screening, including assessment and treatment of screen-positive cases. Eighty five per cent of screening programmes have a coordinated screening service and 73% of these felt that they have made significant progress. Eighty-five per cent of screening units use 'call and recall' for appointments and 73.5% of programmes follow the National Screening Committee (NSC) guidance. Although many units worked closely with ophthalmology, further assessment and management of screen positive patients was a cause for concern. The fast-track referral system, to ensure timely and appropriate care, has been difficult to engineer by several programmes. This is demonstrated by 48% of programmes having waiting lists for patients identified as needing further assessment and treatment for retinopathy. Ophthalmology service for people with diabetic retinopathy was provided by a dedicated ophthalmologist in 89.4% of the programmes. Sixty-six per cent of the programmes reported inadequate resources to sustain a high-quality service, while 26% highlighted the lack of infrastructure and 49% lacked information technology (IT) support. In conclusion, progress has been made towards establishing a national screening programme for diabetic retinopathy by individual screening units, with a number of programmes providing a structured retinal screening service. However, programmes face difficulties with resource allocation and compliance with Quality Assurance (QA) standards, especially those which apply to ophthalmology and IT support. Screening programmes need to be resourced adequately to ensure comprehensive coverage and compliance with QA. PMID- 20002487 TI - Prospective and retrospective recording of severe hypoglycaemia, and assessment of hypoglycaemia awareness in insulin-treated Type 2 diabetes. PMID- 20002488 TI - Advantages of insulin pump therapy. PMID- 20002489 TI - Capecitabine-induced severe hypertriglyceridaemia and diabetes: a case report and review of the literature. PMID- 20002490 TI - Effect of combined oral proteases and flavonoid treatment in subjects at risk of Type 1 diabetes. PMID- 20002492 TI - Disasters has informed humanitarian thinking and practice for more than 30 years. PMID- 20002494 TI - A meta-analysis of trait differences between invasive and non-invasive plant species. AB - A major aim in ecology is identifying determinants of invasiveness. We performed a meta-analysis of 117 field or experimental-garden studies that measured pair wise trait differences of a total of 125 invasive and 196 non-invasive plant species in the invasive range of the invasive species. We tested whether invasiveness is associated with performance-related traits (physiology, leaf-area allocation, shoot allocation, growth rate, size and fitness), and whether such associations depend on type of study and on biogeographical or biological factors. Overall, invasive species had significantly higher values than non invasive species for all six trait categories. More trait differences were significant for invasive vs. native comparisons than for invasive vs. non invasive alien comparisons. Moreover, for comparisons between invasive species and native species that themselves are invasive elsewhere, no trait differences were significant. Differences in physiology and growth rate were larger in tropical regions than in temperate regions. Trait differences did not depend on whether the invasive alien species originates from Europe, nor did they depend on the test environment. We conclude that invasive alien species had higher values for those traits related to performance than non-invasive species. This suggests that it might become possible to predict future plant invasions from species traits. PMID- 20002496 TI - Functional characterization of human nucleosome assembly protein 1-like proteins as histone chaperones. AB - Nucleosome Assembly Protein 1 (NAP1) is a highly conserved histone chaperone protein suspected to be involved in the dynamical regulation of the histone H2A H2B hetero-dimer. However, the exact mechanism by which NAP1-like proteins act is currently unknown. In this work, we characterized the biochemical properties of two human NAP1-like proteins, hNAP1L1 and hNAP1L4, including a previously uncharacterized subtype, with the aim of determining their exact mechanistic role. Both hNAP1L1 and hNAP1L4 were found to be localized mainly to the cytoplasm and a minor population of them was suggested to be in the nucleus. Biochemical analyses demonstrated that both hNAP1L1 and hNAP1L4 mediated nucleosome formation. In addition, hNAP1L1 was shown to possess a significantly greater nucleosome disassembly activity than hNAP1L4, suggesting that hNAP1L1 and hNAP1L4 may play distinct roles in the regulation of histone dynamics. Building upon this initial discovery we also found that histone H2A-H2B and various histone H2A variants-H2B dimers were found to associate with both hNAP1L1 and hNAP1L4 in cell extracts. These results suggest that human NAP1-like proteins play overlapping roles in transport and deposition of histone H2A-H2B or H2A variants-H2B dimers on chromatin and nonoverlapping roles in nucleosome disassembly. PMID- 20002497 TI - Multiple roles of PPARalpha in brown adipose tissue under constitutive and cold conditions. AB - Peroxisome proliferator-activated receptor alpha (PPARalpha) is a member of the nuclear receptor family, regulating fatty acid degradation in many organs. Two dimensional SDS-PAGE of brown adipose tissue (BAT) from PPARalpha-null mice produced a higher-density spot. Proteomic analysis indicated that the protein was pyruvate dehydrogenase beta (PDHbeta). To observe PDHbeta regulation in BAT, the organ was stimulated by long-term cold exposure, and the activities of associated enzymes were investigated. Histological and biochemical analyses of BAT showed a significant decrease in the triglyceride content in wild-type mice and some degree of decrease in PPARalpha-null mice on cold exposure. Analyses of molecules related to glucose metabolism showed that the expression of PDHbeta is under PPARalpha-specific regulation, and that glucose degradation ability may decrease on cold exposure. In contrast, analyses of molecules related to fatty acid metabolism showed that numerous PPARalpha/gamma target molecules are induced on cold exposure, and that fatty acid degradation ability in wild-type mice is markedly enhanced and also increases to same degree in PPARalpha-null mice on cold exposure. Thus, this study proposes novel and multiple roles of PPARalpha in BAT. PMID- 20002498 TI - Regulatory mechanism for expression of GPX1 in response to glucose starvation and Ca in Saccharomyces cerevisiae: involvement of Snf1 and Ras/cAMP pathway in Ca signaling. AB - Saccharomyces cerevisiae has three homologues of the glutathione peroxidase gene, GPX1, GPX2, and GPX3. We have previously reported that the expression of GPX3 was constitutive, but that of GPX2 was induced by oxidative stress and CaCl(2), and uncovered the regulatory mechanisms involved. Here, we show that the expression of GPX1 is induced by glucose starvation and treatment with CaCl(2). The induction of GPX1 expression in response to glucose starvation and Ca(2+) was dependent on the transcription factors Msn2 and Msn4 and cis-acting elements [stress response element (STRE)] in the GPX1 promoter. The Ras/cAMP pathway is also involved in the expression of GPX1. We found that Snf1, a Ser/Thr protein kinase, is involved in the glucose starvation- and Ca(2+)-induced expression of GPX1. The activation of Snf1 is accompanied by phosphorylation of Thr(210). We found that the Ca(2+)-treatment as well as glucose starvation causes the phosphorylation of Thr(210) of Snf1 in a Tos3, Sak1, and Elm1 protein kinase dependent manner. As the timing of the initiation of Ca(2+)-induced expression of GPX1 was retarded in an snf1Delta mutant, the activation of Snf1 seems pivotal to the early-stage-response of GPX1 to Ca(2+). PMID- 20002499 TI - High density of weak replication origins in a 75-kb region of chromosome 2 of fission yeast. AB - Using a two-dimensional gel electrophoresis origin mapping technique and cell synchronization, we have studied replication timing and mapped origins in a 75-kb region of chromosome 2 of Schizosaccharomyces pombe. Three of the five mapped origins are moderately active and the other two are very weak. DNA fragments containing the three moderately active origins and one weak origin are ARS positive whereas that containing the other weak origin is ARS-negative. Three ARS elements reported earlier from this region appear to be inactive as chromosomal origins. The centromere-proximal 45 kb of this region replicates earlier than the telomere-proximal 30 kb. A transition from early to late replication occurs within 10 kb of the chromosomally inactive ars727, suggesting a possible role of the previously reported late-replication-enforcing region in determining chromosomal replication timing of the region. These results in conjunction with those from some other studies suggest that, in S. pombe, the actual number of potential origins may be significantly higher than previously detected in many genome-wide studies, and the relationship between ARS activity and chromosomal origin activity is not as simple as in Saccharomyces cerevisiae. PMID- 20002500 TI - Prevalence and factors associated with renal impairment in HIV-infected patients, ANRS C03 Aquitaine Cohort, France. AB - OBJECTIVES: The aims of the present study were to estimate the prevalence of renal impairment (RI) among HIV-infected adult patients and to investigate the associated factors. METHODS: A cross-sectional survey was conducted in a French hospital-based cohort. Clearance of creatinine (CC) was calculated using the Cockcroft-Gault formula. Four stages of RI were defined: mild (60-90 mL/min), moderate (30-60), severe (15-30) and end stage (<15). Logistic regression models were used to investigate factors associated with RI. RESULTS: The male/female ratio of the 2588 patients enrolled was 3:1 and the median age was 42 years. At the time of assessment of CC, the median CD4 count was 430 cells/microL and HIV plasma viral load (VL) was<50 copies/mL in 60%. The overall prevalence of RI was 39.0%: 34.2% mild, 4.4% moderate, 0.3% severe and 0.2% end-stage. Mild RI was associated with female gender [odds ratio (OR)=3.3: 95% CI 2.6-4.3)], age >50 years (OR=9.8: 7.4-13.0) and 40-50 years (OR=1.9: 1.5-2.4), body mass index (BMI) <22 kg/m(2) (OR=3.3: 2.7-4.3) and tenofovir exposure (OR=1.4: 1.0-1.9 for <1 year and OR=1.5: 1.2-2.0 for >1 year). Advanced RI (CC <60 mL/min) was associated with age >50 years (OR=5.6: 2.9-10.9) and 40-50 years (OR=2.2: 1.1-1.4), BMI <22 kg/m(2) (OR=1.5: 1.0-2.4), hypertension (OR=2.5: 1.4-2.5) and indinavir (IDV) exposure >1 year (OR=2.3: 1.5-3.6). CONCLUSION: This survey confirms the high prevalence of RI in HIV-infected patients and indicates the importance of the investigation of renal function especially in women, older patients, those with a low BMI or treated with tenofovir or IDV. PMID- 20002501 TI - Monitoring and impact of fluconazole serum and cerebrospinal fluid concentration in HIV-associated cryptococcal meningitis-infected patients. AB - OBJECTIVES: The aim of the present study was to assess fluconazole pharmacokinetic measures in serum and cerebrospinal fluid (CSF); and the correlation of these measures with clinical outcomes of invasive fungal infections. METHODS: A randomized trial was conducted in HIV-infected patients receiving three different regimens of fluconazole plus amphotericin B (AmB) for the treatment of cryptococcal meningitis. Regimens included fluconazole 400 mg/day+AmB (AmB+Fluc400) or fluconazole 800 mg/day+AmB (AmB+Fluc800) (14 days followed by fluconazole alone at the randomized dose for 56 days); or AmB alone for 14 days followed by fluconazole 400 mg/day for 56 days. Serum (at 24 h after dosing) and CSF samples were taken at baseline and days 14 and 70 (serum only) for fluconazole measurement, using gas-liquid chromatography. RESULTS: Sixty-four treated patients had fluconazole measurements: 11 in the AmB group, 12 in the AmB+Fluc400 group and 41 in the AmB+Fluc800 group. Day 14 serum concentration geometric means were 24.7 mg/L for AmB+Fluc400 and 37.0 mg/L for AmB+Fluc800. Correspondingly, CSF concentration geometric means were 25.1 mg/L and 32.7 mg/L. Day 14 Serum and CSF concentrations were highly correlated with AmB+Fluc800 (P<0.001, r=0.873) and AmB+Fluc400 (P=0.005, r=0.943). Increased serum area under the curve (AUC) appears to be associated with decreased mortality at day 70 (P=0.061, odds ratio=2.19) as well as with increased study composite endpoint success at days 42 and 70 (P=0.081, odds ratio=2.25 and 0.058, 2.89, respectively). CONCLUSION: High fluconazole dosage (800 mg/day) for the treatment of HIV-associated cryptococcal meningitis was associated with high serum and CSF fluconazole concentration. Overall, high serum and CSF concentration appear to be associated with increased survival and primary composite endpoint success. PMID- 20002502 TI - Development and initial validation of a brief screener for focused HIV prevention efforts. AB - OBJECTIVES: The public health response to the spread of HIV relies on behavioural changes, especially reductions in sexual and drug-use-related transmission risk behaviours (TRBs). While understanding the factors that dispose people towards risky behaviours is important scientifically, it can be difficult to distil the many predictors of sexual risk behaviours into a useful clinical tool for focused prevention efforts. Our goal was to evaluate the extent to which known predictors of sexual TRBs (self-efficacy, treatment optimism, engagement with medical care, awareness of risky behaviours, substance use, and relevant behavioural and socio demographic characteristics) combined with additional attitude-related assessments to identify those who had engaged in recent sexual TRBs and may therefore be at risk of additional TRBs. METHODS: In this study, we analysed data on beliefs and behaviours related to sex, substance use, HIV prevention and other relevant factors for 280 patients at a publicly funded HIV/AIDS clinic in Seattle. All participants completed a baseline audio computer-assisted self interview (ACASI) as part of a larger trial focused on reducing TRBs. RESULTS: Our multivariate model yielded three screening questions that could prove effective in identifying HIV-positive patients in need of focused prevention resources. CONCLUSIONS: The resulting screener holds promise as a brief and easily deployed tool that can be used by providers regardless of access to ACASI technology. Additional validation is needed and longitudinal evaluation is currently in progress. PMID- 20002503 TI - Editorial Comment to Post-transplant lymphoproliferative disorder following renal transplantation: a single-center experience over 40 years. PMID- 20002504 TI - Post-transplant lymphoproliferative disorder following renal transplantation: a single-center experience over 40 years. AB - OBJECTIVES: To investigate post-transplant lymphoproliferative disorder (PTLD) following renal transplantation at our institution. METHODS: Medical records of 631 patients who underwent renal transplantation at Osaka University Hospital between March 1965 and December 2008 were reviewed. RESULTS: PTLD following renal transplantation was detected in 10 patients (five men, five women; mean age at transplantation, 38.5 years). Mean duration from renal transplantation to the onset of PTLD was 7.1 years (range, 5 months to 18 years, 9 months). Mean duration of observation was 3.9 years from the onset of PTLD. Immunosuppressant therapy comprised multidrug combination therapy, including cyclosporine in six patients and tacrolimus in four patients. In addition to a reduction in the immunosuppressant dose, which was performed in all patients, PTLD was treated with surgery in seven patients, radiotherapy in two patients, rituximab in five patients, and cytotoxic chemotherapy in four patients. A complete remission in eight patients and progressive disease in two were observed. At last follow up, seven patients were alive and five patients had functioning grafts. CONCLUSIONS: The incidence of PTLD following renal transplantation at our institution is 1.6% with onset occurring more than 5 years after transplantation in five patients. Consequently, with long-term renal graft survival now feasible, attention must be paid to detecting late-onset PTLD. PMID- 20002505 TI - Effect of a statewide fall prevention program on incidence of femoral fractures in residents of long-term care facilities. AB - OBJECTIVES: To evaluate the effect of a statewide dissemination of a modified evidence-based fall prevention program on incidence of femoral fractures in nursing homes. DESIGN: Observational study of a staged implementation of a successful fall prevention program. SETTING: One thousand three hundred fifty nine nursing homes in two federal states in the south of Germany. PARTICIPANTS: Nine thousand seventy-seven residents in the intervention homes and 43,583 residents in control homes from the same and a different federal state. INTERVENTION: Staff education on fall prevention, advice on environmental adaptations, and progressive strength and balance training over at least 1 year. MEASUREMENTS: Incident femoral fractures. RESULTS: One thousand five hundred eighteen femoral fractures occurred in the total study population during the intervention period. The crude incidence rate of femoral fractures was 39.5/1,000 person-years in residents from the intervention homes and 40.9 and 39.7/1,000 person-years in residents from two sets of control homes. In a multivariate model, there was no evidence of an effect of the fall prevention program on incidence of femoral fracture when compared with control homes from the same federal state (adjusted hazard ratio (AHR)=0.96, 95% confidence interval (CI)=0.83-1.11) or from a different federal state (AHR=1.00, 95% CI=0.86-1.16). CONCLUSION: The statewide dissemination of a multifactorial fall prevention program did not appear to reduce the burden of femoral fractures in residents of nursing homes. PMID- 20002506 TI - Implementing group medical visits for older adults at group health cooperative. AB - In a pair of randomized controlled trials in the Kaiser Permanente delivery system in Colorado in the 1990s, group visits for older adults (monthly non disease-specific group medical appointments for a cohort of patients led by primary care teams) were proven to reduce costs, decrease hospitalizations, and improve patient and provider satisfaction. As part of a translational effort, this group visit intervention was replicated in a delivery system in Seattle, Washington, and the log of total healthcare costs was measured in the first year of the intervention. Utilization and patient and physician satisfaction were secondary outcomes. For the cost and utilization analysis, a retrospective case control design compared 221 case patients aged 65 and older with high outpatient usage in the previous 18 months with 1,015 control patients selected randomly from clinics not participating in the intervention. Controls were matched to cases on the number of primary care visits in the prior 18 months. Total costs were not statistically different for intervention patients and controls ($8,845 vs $10,288, P=.11), nor were there statistically significant differences in utilization, including hospital admissions and outpatient visits, but patient and provider satisfaction with the intervention was high. This translational effort did not demonstrate the cost savings of the original efficacy trials. Possible explanations for these divergent results may have to do with differences between those who participated and differences between the two delivery systems. PMID- 20002507 TI - Training nursing staff to recognize depression in home healthcare. AB - OBJECTIVES: To describe the implementation and acceptability of the TRaining In the Assessment of Depression (TRIAD) intervention, which has been tested in a randomized trial. The primary aim of TRIAD is to improve the ability of homecare nurses to detect depression in medically ill, older adult homecare patients. DESIGN: Description of the important components of TRIAD, its implementation, and evaluation results from nurse surveys. SETTING: Three certified home healthcare agencies in Westchester County, New York. PARTICIPANTS: Thirty-six homecare nurses. INTERVENTION: Participants randomly assigned to TRIAD (n=17) were provided with the opportunity to observe and practice patient interviewing. The approach focused on clinically meaningful identification of the two "gateway" symptoms of depression and is consistent with the newly revised Medicare mandatory Outcome and Assessment Information Set (OASIS-C). Control group participants (n=19) received no training beyond that which agencies may have provided routinely. MEASUREMENTS: Baseline and 1-year nurse confidence in depression detection, and postintervention acceptability ratings of the TRIAD intervention. RESULTS: Participants randomized to the TRIAD intervention reported a statistically significant increase in confidence in assessing for depression mood (P<.001), whereas the usual care group's confidence remained unchanged (P=.34) 1 year later. CONCLUSION: An educational program designed to improve depression detection by giving nurses the skills and confidence to integrate depression assessment into the context of routine care can be successfully implemented with homecare agency support. The authors discuss the intervention in terms of OASIS-C and the "real world" realities of intervention implementation. PMID- 20002508 TI - Higher level of systemic C-reactive protein is independently predictive of coronary heart disease in older community-dwelling adults: the three-city study. AB - OBJECTIVES: To assess the association between systemic C-reactive protein (CRP) and incident coronary heart disease (CHD) in community-dwelling elderly people. DESIGN: A French population-based multicenter prospective cohort study. SETTING: Three cities in France: Bordeaux in the southwest, Dijon in the northeast, and Montpellier in the southeast. PARTICIPANTS: After 4 years of follow-up, a case cohort study was designed including 1,004 subjects randomly selected from the initial cohort of 9,294 subjects free of CHD at baseline and 174 subjects who developed first CHD events during follow-up. MEASUREMENTS: Hazard ratios (HRs) were estimated using a Cox proportional hazard model adapted for the case-cohort design using a CRP level less than 1 mg/L as the reference category. RESULTS: Of the random sample, 24.3% had a CRP level less than 1.0 mg/L, 45.8% had a CRP level of 1.0 to 2.9 mg/L, and 29.9% had a CRP level of 3.0 to 10.0 mg/L. The HRs for CHD, adjusted for age, sex, and study center, were 1.69 (95% confidence interval (CI)=1.04-2.75) for CRP from 1.0 to 2.9 mg/L and 2.32 (95% CI=1.41-3.82) for CRP from 3.0 to 10.0 mg/L (P for trend <.001). After additional adjustment for smoking, body mass index, diabetes mellitus, systolic blood pressure, high density lipoprotein cholesterol, low-density lipoprotein cholesterol, triglycerides, statin use, and antihypertensive treatment, a baseline CRP of 3.0 to 10.0 mg/L remained associated with risk of CHD (HR=1.87, 95% CI=1.09-3.25), although CRP did not improve the discriminative ability of a predicting model based on traditional risk factors (receiver operating characteristic curves from 0.740 to 0.749). CONCLUSION: CRP is an independent CHD risk marker but does not improve CHD risk prediction in community-dwelling elderly people. PMID- 20002509 TI - The effectiveness of inpatient geriatric evaluation and management units: a systematic review and meta-analysis. AB - OBJECTIVES: To examine how geriatric evaluation and management units (GEMUs) are organized and to examine the effectiveness of admission on a GEMU. DESIGN: Systematic review and meta-analysis based on literature search of multiple databases and the references lists of all identified articles and by contacting authors. SETTING: GEMUs. PARTICIPANTS: Elderly people admitted to a GEMU. MEASUREMENTS: Quality of the studies was assessed on 10 criteria. The outcome parameters were mortality, institutionalization, functional decline, readmission, and length of stay at different follow-up points. A random-effects meta-analysis was performed using Hedges' gu and variance of relative risk (RR). RESULTS: GEMUs are organized in a heterogeneous way and the included studies gave no thorough description of comprehensive geriatric assessment (CGA). Involvement of a multidisciplinary team was a key element in all GEMUs. The individual trials showed that admission to a GEMU has one or more favorable effects on the outcomes of interest, with two significant effects in the meta-analysis: less functional decline at discharge from the GEMU (RR=0.87, 95% confidence interval (CI)=0.77 0.99; P=.04) and a lower rate of institutionalization 1 year after discharge (RR=0.78, CI=0.66-0.92; P=.003). For the other outcomes in the meta-analysis, a GEMU did not induce significantly different outcomes than usual care. CONCLUSION: This meta-analysis shows a significant effect in favor of the GEMU group on functional decline at discharge and on institutionalization after 1 year. There is heterogeneity between the studies, poor quality of some randomized controlled trials, and shortage of information about CGA. Multidisciplinary CGA offered in a GEMU may add value to the care for frail older persons admitted to the hospital, but the limitations confirm the need for well-designed studies using explicit CGA and more-structured and -coherent assessment instruments such as the Minimum Data Set Resident Assessment Instrument. PMID- 20002510 TI - An evaluation of an adapted U.S. model of pharmaceutical care to improve psychoactive prescribing for nursing home residents in northern ireland (fleetwood northern ireland study). AB - OBJECTIVES: To test the effect of an adapted U.S. model of pharmaceutical care on prescribing of inappropriate psychoactive (anxiolytic, hypnotic, and antipsychotic) medications and falls in nursing homes for older people in Northern Ireland (NI). DESIGN: Cluster randomized controlled trial. SETTING: Nursing homes randomized to intervention (receipt of the adapted model of care; n=11) or control (usual care continued; n=11). PARTICIPANTS: Residents aged 65 and older who provided informed consent (N=334; 173 intervention, 161 control). INTERVENTION: Specially trained pharmacists visited intervention homes monthly for 12 months and reviewed residents' clinical and prescribing information, applied an algorithm that guided them in assessing the appropriateness of psychoactive medication, and worked with prescribers (general practitioners) to improve the prescribing of these drugs. The control homes received usual care. MEASUREMENTS: The primary end point was the proportion of residents prescribed one or more inappropriate psychoactive medicine according to standardized protocols; falls were evaluated using routinely collected falls data mandated by the regulatory body for nursing homes in NI. RESULTS: The proportion of residents taking inappropriate psychoactive medications at 12 months in the intervention homes (25/128, 19.5%) was much lower than in the control homes (62/124, 50.0%) (odds ratio=0.26, 95% confidence interval=0.14-0.49) after adjustment for clustering within homes. No differences were observed at 12 months in the falls rate between the intervention and control groups. CONCLUSION: Marked reductions in inappropriate psychoactive medication prescribing in residents resulted from pharmacist review of targeted medications, but there was no effect on falls. PMID- 20002511 TI - The poor outcome of ischemic stroke in very old people: a cohort study of its determinants. AB - OBJECTIVES: To assess how much of the excess risk of poor outcome from stroke in people aged 80 and older aging per se explains, independent of other prognostic determinants. DESIGN: Cohort, observational. SETTING: University hospital. PARTICIPANTS: One thousand five hundred fifty-five patients with first-ever ischemic stroke consecutively referred to an in-hospital Clinical Pathway program were studied. MEASUREMENTS: The relationship between age and 1-month outcome (death, disability (modified Rankin Scale 3-5), and poor outcome (modified Rankin Scale 3-6)) was assessed, with adjustment for several prognostic factors. RESULTS: Six hundred twelve patients aged 80 and older showed worse outcome after 1 month than those who were younger, in terms of mortality (19% vs 5%, hazard ratio (HR)=3.85, 95% confidence interval (CI)=2.8-5.4) and disability (51% vs 33%, odds ratio (OR)=3.16, 95% CI=2.5-4.0), although in multivariate models, the adjusted HR for mortality decreased to 1.47 (95% CI=1.0-2.16) and the ORs for disability and poor outcome decreased to 1.76 (95% CI=1.32-2.3.) and 1.83 (95% CI=137-2.43), respectively. Stroke severity, the occurrence of at least one medical complication, and premorbid disability explained most of the risk excess in the oldest-old. CONCLUSION: Stroke outcome is definitely worse in very old people, and most of the excess risk of death and disability is attributable to the higher occurrences of the most-severe clinical stroke syndromes and of medical complications in the acute phase. These represent potential targets for preventive and therapeutical strategies specifically for elderly people. PMID- 20002512 TI - Hearing handicap predicts the development of depressive symptoms after 3 years in older community-dwelling Japanese. AB - OBJECTIVES: To examine the association between hearing handicap and depressive symptoms in older community-dwelling Japanese. DESIGN: Community-based cohort study. SETTING: Kurabuchi Town, Gunma Prefecture, Japan. PARTICIPANTS: Five hundred eighty residents (261 men, 319 women) aged 65 and older without depressive symptoms. MEASUREMENTS: In a baseline examination performed in 2005/06, participants answered the 10-item screening version of the Hearing Handicap Inventory for Elderly (HHIE-S). They were divided into two groups according to their scores: a group with no hearing handicap (HHIE-S scores of < or =8) and a hearing handicap group (HHIE-S scores of > or =10). The Geriatric Depression Scale was used to identify depressive symptoms in face-to-face home visit interviews conducted in 2008, and the association between hearing handicap and depressive symptoms was assessed using logistic regression. RESULTS: The incidence of depressive symptoms was 19.6% in the group with a hearing handicap and 8.0% in the group without a hearing handicap. When compared with the subjects without hearing handicap, subjects with a hearing handicap had a multiadjusted odds ratio of depressive symptoms of 2.45 (95% confidence interval=1.26-4.77). The association remained significant even when hearing impairment measured with pure-tone audiometry was added to the multiadjusted model. CONCLUSION: A hearing handicap can predict future depressive symptoms in older community-dwelling people. PMID- 20002513 TI - Volume of home- and community-based Medicaid waiver services and risk of hospital admissions. AB - OBJECTIVES: To evaluate whether type and volume of Medicaid Home- and Community Based Services (HCBS) waiver program are associated with risk of hospitalization and whether this association changes over time. DESIGN: Prospective. SETTING: Indiana Medicaid claims data from June 2001 to December 2004. PARTICIPANTS: Medicaid recipients (N=1,354) who enrolled in the Aged and Disabled waiver program between January 2002 and June 2004. MEASUREMENTS: Time to hospital admission since enrollment in the HCBS waiver program, adjusted for demographics, comorbidities, prior use of health services, and volume of HCBS received, including attendant care, homemaking, and home-delivered meals. RESULTS: A greater volume of attendant care, homemaking services, and home-delivered meals was associated with a lower risk of hospitalization. This effect diminished over time for attendant care and homemaking. The risk of hospitalization for subjects receiving 5 hours of attendant care per month was 54% (hazard ratio (HR)=0.46, 95% confidence interval (CI)=0.38-0.57) lower during the first month of enrollment and 20% lower by Month 10 (HR=0.80, 95% CI=0.73-0.88) than for those receiving no attendant care. CONCLUSION: Greater volume of HCBS services was associated with lower risk of hospitalization. The findings highlight the potential importance of assessing and monitoring the volume of HCBS patients receive. PMID- 20002514 TI - Fecal incontinence in elderly Koreans. AB - OBJECTIVES: To estimate the prevalence and correlates of fecal incontinence (FI) and its effect on quality of life in ambulatory elderly people in Korea. DESIGN: Cross-sectional, convenience sample-based survey. SETTING: Twenty-seven senior citizen centers and two health clinics in two cities of Korea. PARTICIPANTS: Nine hundred eighty-one relatively healthy and ambulatory community-dwelling people aged 60 and older. MEASUREMENTS: Data were collected through in-person interviews with a structured questionnaire. Multivariate logistic regression analysis was used to determine independent risk factors for FI. RESULTS: The prevalence of FI was 15.5%. FI was significantly associated with lower quality of life (Medical Outcomes Study 36-item Short-Form Survey) for physical and mental health. In men, FI was significantly associated with urinary incontinence (odds ratio (OR)=4.89, 95% confidence interval (CI)=2.45-9.77), hemorrhoids (OR=4.66, 95% CI=1.67 12.97), and poor self-perceived health status (P for trend=.02). In women, FI was associated with urinary incontinence (OR=2.91, 95% CI=1.76-4.81), diabetes mellitus (OR=2.04, 95% CI=1.24-3.37), hemorrhoids (OR=2.99, 95% CI=1.31-6.83), and infrequent dietary fiber intake (P for trend=.02). CONCLUSION: FI is prevalent in elderly Koreans and has a profound effect on quality of life. Physicians should closely screen for FI in elderly patients with certain risk factors and evaluate to control these potentially preventable or modifiable factors. PMID- 20002515 TI - Chronic hyperammonemia induces tonic activation of NMDA receptors in cerebellum. AB - Reduced function of the glutamate--nitric oxide (NO)--cGMP pathway is responsible for some cognitive alterations in rats with hyperammonemia and hepatic encephalopathy. Hyperammonemia impairs the pathway in cerebellum by increasing neuronal nitric oxide synthase (nNOS) phosphorylation in Ser847 by calcium calmodulin-dependent protein kinase II (CaMKII), reducing nNOS activity, and by reducing nNOS amount in synaptic membranes, which reduces its activation following NMDA receptors activation. The reason for increased CaMKII activity in hyperammonemia remains unknown. We hypothesized that it would be as a result of increased tonic activation of NMDA receptors. The aims of this work were to assess: (i) whether tonic NMDA activation receptors is increased in cerebellum in chronic hyperammonemia in vivo; and (ii) whether this tonic activation is responsible for increased CaMKII activity and reduced activity of nNOS and of the glutamate--NO--cGMP pathway. Blocking NMDA receptors with MK-801 increases cGMP and NO metabolites in cerebellum in vivo and in slices from hyperammonemic rats. This is because of reduced phosphorylation and activity of CaMKII, leading to normalization of nNOS phosphorylation and activity. MK-801 also increases nNOS in synaptic membranes and reduces it in cytosol. This indicates that hyperammonemia increases tonic activation of NMDA receptors leading to reduced activity of nNOS and of the glutamate--NO--cGMP pathway. PMID- 20002516 TI - Differential modulation of mu-opioid receptor signaling to adenylyl cyclase by regulators of G protein signaling proteins 4 or 8 and 7 in permeabilised C6 cells is Galpha subtype dependent. AB - Regulators of G protein signaling (RGS) proteins act as GTPase-accelerating protein to negatively modulate G protein signaling and are defined by a conserved RGS domain with considerable amino acid diversity. To determine the effects of specific, purified RGS proteins on mu-opioid signaling, C6 cells stably expressing a mu-opioid receptor were rendered permeable to proteins by treatment with digitonin. Mu-opioid inhibition of forskolin-stimulated adenylyl cyclase by [D-Ala(2),N-Me-Phe(4),Gly-ol]-enkephalin (DAMGO), a mu-specific opioid peptide, remained fully intact in permeabilized cells. Purified RGS domain of RGS4 added to permeabilized cells resulted in a twofold loss in DAMGO potency but had no effect in cells expressing RGS-insensitive G proteins. The inhibitory effect of DAMGO was reduced to the same extent by purified RGS4 and RGS8. In contrast, the RGS domain of RGS7 had no effect and inhibited the action of RGS8 as a result of weak physical association with Galphai2 and minimal GTPase-accelerating protein activity in C6 cell membranes. These data suggest that differences in conserved RGS domains of specific RGS proteins contribute to differential regulation of opioid signaling to adenylyl cyclase and that a permeabilized cell model is useful for studying the effects of specific RGS proteins on aspects of G protein coupled receptor signaling. PMID- 20002517 TI - Cyclic GMP phosphodiesterase inhibition alters the glial inflammatory response, reduces oxidative stress and cell death and increases angiogenesis following focal brain injury. AB - Recent evidence obtained in cultured glial cells indicates that cGMP-mediated pathways regulate cytoskeleton dynamics, glial fibrillary acidic protein expression and motility in astrocytes, as well as inflammatory gene expression in microglia, suggesting a role in the regulation of the glial reactive phenotype. The aim of this work was to examine if cGMP regulates the glial inflammatory response in vivo following CNS damage caused by a focal cryolesion onto the cortex in rats. Results show that treatment with the cGMP phosphodiesterase inhibitor zaprinast (10 mg/kg i.p.) 2 h before and 24 and 48 h after the lesion results 3 days post-lesion in notably enhanced astrogliosis manifested by increased glial fibrillary acidic protein immunoreactivity and protein levels around the lesion. In contrast, zaprinast decreased the number of round/ameboid lectin-positive cells and the expression of the activated microglia/macrophage markers Iba-1 and CD11b indicating decreased recruitment and activation of these cells. This altered inflammatory response is accompanied by a decrease in protein oxidative stress, apoptotic cell death and neuronal degeneration. In addition, zaprinast enhanced angiogenesis in the lesioned cortex probably as a result of vascular endothelial growth factor expression in reactive astrocytes. These results suggest that regulation of the glial inflammatory response may contribute to the reported neuroprotective effects of cGMP-phosphodiesterase inhibitors in brain injury. PMID- 20002518 TI - Circadian variations of prostaglandin E2 and F2 alpha release in the golden hamster retina. AB - Circadian variations of prostaglandin E2 and F2alpha release were examined in the golden hamster retina. Both parameters showed significant diurnal variations with maximal values at midnight. When hamsters were placed under constant darkness for 48 h, the differences in prostaglandin release between subjective mid-day and subjective midnight persisted. Western blot analysis showed that cyclooxygenase (COX)-1 levels were significantly higher at midnight than at mid-day, and at subjective midnight than at subjective mid-day, whereas no changes in COX-2 levels were observed among these time points. Immunohistochemical studies indicated the presence of COX-1 and COX-2 in the inner (but not outer) retina. Circadian variations of retinal prostaglandin release were also assessed in suprachiasmatic nuclei (SCN)-lesioned animals. Significant differences in retinal prostaglandin release between subjective mid-day and subjective midnight were observed in SCN-lesioned animals. These results indicate that hamster retinal prostaglandin release is regulated by a retinal circadian clock independent from the SCN. Thus, the present results suggest that the prostaglandin/COX-1 system could be a retinal clock output or part of the retinal clock mechanism. PMID- 20002519 TI - AAV-mediated chronic over-expression of SNAP-25 in adult rat dorsal hippocampus impairs memory-associated synaptic plasticity. AB - Long-term memory is formed by alterations in glutamate-dependent excitatory synaptic transmission, which is in turn regulated by synaptosomal protein of 25 kDa (SNAP-25), a key component of the soluble N-ethylmaleimide-sensitive factor attachment protein receptor complex essential for exocytosis of neurotransmitter filled synaptic vesicles. Both reduced and excessive SNAP-25 activity has been implicated in various disease states that involve cognitive dysfunctions such as attention deficit hyperactivity disorder, schizophrenia and Alzheimer's disease. Here, we over-express SNAP-25 in the adult rat dorsal hippocampus by infusion of a recombinant adeno-associated virus vector, to evaluate the consequence of late adolescent-adult dysfunction of the soluble N-ethylmaleimide-sensitive factor attachment protein receptor protein in the absence of developmental disruption. We report a specific and significant increase in the levels of extracellular glutamate detectable by microdialysis and a reduction in paired-pulse facilitation in the hippocampus. In addition, SNAP-25 over-expression produced cognitive deficits, delaying acquisition of a spatial map in the water maze and impairing contextual fear conditioning, both tasks known to be dorsal hippocampal dependent. The high background transmission state and pre-synaptic dysfunction likely result in interference with requisite synapse selection during spatial and fear memory consolidation. Together these studies provide the first evidence that excess SNAP-25 activity, restricted to the adult period, is sufficient to mediate significant deficits in the memory formation process. PMID- 20002520 TI - Effects of 3,4-methylenedioxymethamphetamine (MDMA) on serotonin transporter and vesicular monoamine transporter 2 protein and gene expression in rats: implications for MDMA neurotoxicity. AB - 3,4-Methylenedioxymethamphetamine (MDMA; 'Ecstasy') is a popular recreational drug used worldwide. This study aimed to determine the effects of this compound on the expression of nerve terminal serotonergic markers in rats. Experiment 1 investigated MDMA-induced changes in levels of the serotonin transporter (SERT) and the vesicular monoamine transporter 2 (VMAT-2) in the hippocampus, a region with sparse dopaminergic innervation, after lesioning noradrenergic input with N (2-chloroethyl)-N-ethyl-2-bromobenzylamine (DSP-4). Adult male Sprague-Dawley rats were administered 100 mg/kg DSP-4 or saline 1 week prior to either an MDMA (10 mg/kg x 4) or saline binge. Two weeks following the binge treatment, the DSP 4/MDMA group unexpectedly showed little change in hippocampal VMAT-2 protein expression compared with DSP-4/Saline controls, despite large reductions in SERT levels in all regions examined in the MDMA-treated animals. Furthermore, animals treated with binge MDMA (Experiment 2) showed a striking decrease in SERT gene expression (and a lesser effect on VMAT-2) measured by quantitative RT-PCR in pooled dorsal and median raphe tissue punches, when compared with saline-treated controls. These results demonstrate that MDMA causes substantial regulatory changes in the expression of serotonergic markers, thus questioning the need to invoke distal axotomy as an explanation of MDMA-related serotonergic deficits. PMID- 20002521 TI - The neuroprotective mechanism of 1-(R)-aminoindan, the major metabolite of the anti-parkinsonian drug rasagiline. AB - The anti-parkinsonian drug, rasagiline [N-propargyl-1-(R)-aminoindan; Azilect(R)], is a secondary cyclic benzylamine and indane derivative, which provides irreversible, potent monoamine oxidase-B (MAO-B) inhibition and possesses neuroprotective and neurorestorative activities. A prospective clinical trial has shown that rasagiline confers significant symptomatic improvement and demonstrated alterations in Parkinson's disease progression. Rasagiline is primarily metabolized by hepatic cytochrome P-450 to form its major metabolite, 1 (R)-aminoindan, a non-amphetamine, weak reversible MAO-B inhibitor compound. Recent studies indicated the potential neuroprotective effect of 1-(R) aminoindan, suggesting that it may contribute to the overall neuroprotective and antiapoptotic effects of its parent compound, rasagiline. This review article briefly highlights the molecular mechanisms underlying the neuroprotective properties of the active metabolite of rasagiline, 1-(R)-aminoindan, supporting the valuable potential of rasagiline for disease modification. PMID- 20002522 TI - Endogenous purinergic signaling is required for osmotic volume regulation of retinal glial cells. AB - Intense neuronal activity in the sensory retina is associated with a volume increase of neuronal cells (Uckermann et al., J. Neurosci. 2004, 24:10149) and a decrease in the osmolarity of the extracellular space fluid (Dmitriev et al., Vis. Neurosci. 1999, 16:1157). Here, we show the existence of an endogenous purinergic mechanism that prevents hypoosmotic swelling of retinal glial (Muller) cells in mice. In contrast to the cells from wild-type mice, hypoosmotic stress induced rapid swelling of glial cell somata in retinal slices from mice deficient in P2Y(1), adenosine A(1) receptors, or ecto-5'-nucleotidase (CD73). Consistently, glial cell bodies in retinal slices from wild-type mice displayed osmotic swelling when P2Y(1) or A(1) receptors, or CD73, were pharmacologically blocked. Exogenous ATP, UTP, and UDP inhibited glial swelling in retinal slices, while the swelling of isolated glial cells was prevented by ATP but not by UTP or UDP, suggesting that uracil nucleotides indirectly regulate the glial cell volume via activation of neuronal P2Y(4/6) and neuron-to-glia signaling. It is suggested that autocrine/paracrine activation of purinergic receptors and enzymes is crucially involved in the regulation of the glial cell volume. PMID- 20002523 TI - Inhibition of Abeta aggregation and neurotoxicity by the 39-kDa receptor associated protein. AB - Aggregation of beta-amyloid protein (Abeta) to form oligomers is considered to be a key step in generating neurotoxicity in the Alzheimer's disease brain. Agents that bind to Abeta and inhibit oligomerization have been proposed as Alzheimer's disease therapeutics. In this study, we investigated the binding of fluorescein labeled Abeta(1-42) (FluoAbeta(1-42)) to SH-SY5Y neuroblastoma cells and examined the effect of the 39-kDa receptor-associated protein (RAP), on the Abeta cell interaction. FluoAbeta(1-42) bound to the cells in a punctate pattern. Surprisingly, when RAP was added to the incubations, FluoAbeta(1-42) and RAP were found to be co-localized on the cell surface, suggesting that RAP and Abeta may bind to each other. Experiments using the purified proteins confirmed that a RAP Abeta complex was stable and resistant to sodium dodecyl sulfate. RAP also inhibited Abeta oligomerization. We next examined whether RAP could inhibit the neurotoxic effects of Abeta. Addition of Abeta(1-42) to SH-SY5Y cells caused an increase in intracellular Ca2+ that was inhibited by treatment of the Abeta peptide with RAP. RAP also blocked an Abeta-induced inhibition of long-term memory consolidation in 1-day-old chicks. This study demonstrates that RAP binds to Abeta and is an inhibitor of the neurotoxic effects of Abeta. PMID- 20002524 TI - Stress and GABA receptors. AB - GABA(A) receptors are sensitive to subtle changes in the environment in both early-life and adulthood. These neurochemical responses to stress in adulthood are sex-dependent. Acute stress induces rapid changes in GABA(A) receptors in experimental animals, with the direction of the changes varying according to the sex of the animals and the stress-paradigm studied. These rapid alterations are of particular interest as they provide an example of fast neurotransmitter system plasticity that may be mediated by stress-induced increases in neurosteroids, perhaps via effects on phosphorylation and/or receptor trafficking. Interestingly, some studies have also provided evidence for long-lasting changes in GABA(A) receptors as a result of exposure to stressors in early-life. The short- and long-term stress sensitivity of the GABAergic system implicates GABA(A) receptors in the non-genetic etiology of psychiatric illnesses such as depression and schizophrenia in which stress may be an important factor. PMID- 20002525 TI - DNA methylation regulates adenosine A(2A) receptor cell surface expression levels. AB - Adenosine A(2A) receptors (A(2A)Rs) appear to play important roles in inflammation and in certain diseases of the nervous system. Pharmacological modulation of A(2A)Rs is particularly useful in Parkinson's disease and has been tested in schizophrenia. However, little is known about the regulation of A(2A)R gene (ADORA2A). A bioinformatic analysis revealed the presence of three CpG islands in the 5' UTR region of human ADORA2A. Next, HeLa, SH-SY5Y and U87-MG cells were treated for 48 h with 5 muM 5-azacytidine (Aza). Increased A(2A)R levels were demonstrated in HeLa and SH-SY5Y cells when compared with non-treated cells. No modifications were seen in U87-MG cells. The increased A(2A)R mRNA and protein levels were accompanied by a loss of DNA methylation pattern in HeLa and SH-SY5Y cells, as measured with the SEQUENOM MassArray platform. The Aza treatment also reduced the affinity of a methyl-CpG-binding protein for ADORA2A by quantitative chromatin immunoprecipitation in HeLa cells. Interestingly, A(2A)R levels were reduced by S-adenosyl-l-methionine treatment in U87-MG and methyl-CpG-binding protein affinity was increased for ADORA2A by quantitative chromatin immunoprecipitation. Therefore, these results show for the first time that DNA methylation plays a role in ADORA2A transcription and, subsequently, in constitutive A(2A)R cell surface levels. PMID- 20002526 TI - Aging-dependent changes of microglial cells and their relevance for neurodegenerative disorders. AB - Among multiple structural and functional brain changes, aging is accompanied by an increase of inflammatory signaling in the nervous system as well as a dysfunction of the immune system elsewhere. Although the long-held view that aging involves neurocognitive impairment is now dismissed, aging is a major risk factor for neurodegenerative diseases such as Alzheimer;s disease, Parkinson;s disease and Huntington's disease, among others. There are many age-related changes affecting the brain, contributing both to certain declining in function and increased frailty, which could singly and collectively affect neuronal viability and vulnerability. Among those changes, both inflammatory responses in aged brains and the altered regulation of toll like receptors, which appears to be relevant for understanding susceptibility to neurodegenerative processes, are linked to pathogenic mechanisms of several diseases. Here, we review how aging and pro-inflammatory environment could modulate microglial phenotype and its reactivity and contribute to the genesis of neurodegenerative processes. Data support our idea that age-related microglial cell changes, by inducing cytotoxicity in contrast to neuroprotection, could contribute to the onset of neurodegenerative changes. This view can have important implications for the development of new therapeutic approaches. PMID- 20002527 TI - C38, equivalent to BM88, is developmentally expressed in maturing retinal neurons and enhances neuronal maturation. AB - C38 antigen is specifically expressed in neuronal cells of the retina. The purpose of this study was to isolate C38 cDNA and determine its molecular functions. Sequence analysis of C38 cDNA revealed that C38 is equivalent to rat BM88, which has been reported to induce cell-cycle arrest and neuronal differentiation in Neuro2a cells. C38 and Ki67, a marker of proliferating cells, were not colocalized during retinal development. C38 was first detected in the retinal ganglion cells at embryonic day 16, much later than the expression of doublecortin, a marker of immature neurons. Although all the horizontal cells were post-mitotic at this stage, C38 was not detected in horizontal cells until the postnatal period. In addition, C38 over-expression did not induce neuronal differentiation or cell-cycle arrest of pluripotent P19 embryonal carcinoma cells. Instead, C38 promoted maturation during neuronal differentiation of P19 embryonal carcinoma cells by down-regulating Oct-3, a pluripotent cell marker and enhancing the expressions of positive regulators of neurogenesis. In conclusion, during retinal development, C38 is first expressed in post-mitotic retinal neurons and is up-regulated during their maturation. C38 does not induce neuronal competence in pluripotent cells, but does promote maturation in already committed neuronal cells. PMID- 20002528 TI - Intermittent hypoxia during sleep induces reactive gliosis and limited neuronal death in rats: implications for sleep apnea. AB - Sleep apnea (SA) can be effectively managed in humans but it is recognized that when left untreated, SA causes long-lasting changes in neuronal circuitry in the brain. Recent neuroimaging studies gave suggested that these neuronal changes are also present even in patients successfully treated for the acute effects of SA. The cellular mechanisms that account for these changes are not certain but animal models of intermittent hypoxia (IH) during sleep have shown neuronal death and impairment in learning and memory. Reactive gliosis has a drastic effect on neuronal survival and circuitry and in this study we examined the neuro-glial response in brain areas affected by SA. Glial and neuronal alterations were analyzed after 1, 3, 5 and 10 days of exposure to IH (8 h/day during the sleep phase, cycles of 6 min each, 10-21% O2) and observed significant astroglial hyperplasia and hypertrophy in parietal brain cortex and hippocampus by studying gliofibrillary acidic protein, Vimentin, S100B and proliferating cell nuclear antigen expression. In addition, altered morphology, reduced dendrite branching and caspase activation were observed in the CA-1 hippocampal and cortical (layers IV-V) pyramidal neurons at short exposure times (1-3 days). Surprisingly, longer exposure to IH reduced the neuronal death rate and increased neuronal branching in the presence of persistent reactive gliosis. Up-regulation of hypoxia inducible factor 1 alpha (HIF-1alpha) and mdr-1, a HIF-1alpha target gene, were observed and increased expression of receptor for advanced end glycated products and its binding partner S100B were also noted. Our results show that a low number of hypoxic cycles induce reactive gliosis and neuronal death whereas continuous exposure to IH cycles reduced the rate of neuronal death and induced neuronal branching on surviving neurons. We hypothesize that HIF-1alpha and S100B glial factor may improve neuronal survival under hypoxic conditions and propose that the death/survival/re-growth process observed here may underlie brain circuitry changes in humans with SA. PMID- 20002529 TI - Influence of factors related to implant stability detected by wireless resonance frequency analysis device. AB - Resonance frequency analysis (RFA) was introduced as a method for measuring implant stability more than a decade ago. Implant stability quotient (ISQ) values obtained using a recently introduced wireless RFA device have made it possible to evaluate stability in a non-invasive technique; however, there are few studies of the factors that affect ISQ values determined using this device. The aim of the present study was to evaluate the association between ISQ values determined by wireless RFA and various factors related to dental implant stability using a pig cortical bone model. Dental implants (Replace) Select Tapered implants) with a length of 10 mm were placed into pig cortical bone samples, then, ISQ values were determined using wireless RFA under various conditions (probe orientation, diameter of implant, insertion torque and peri-implant bone loss). The results of this study showed that ISQ values were not affected by the direction of the probe from parallel to perpendicular to the long axis of the pig bone or to the smart peg. In addition, the diameter of the implant did not have a significant effect on the measured ISQ values. Statistically significant correlations were found between insertion torque and ISQ values (Spearman's test, P < 0.05), and lower ISQ values were observed for deeper peri-implant vertical defects (Mann-Whitney U test, P < 0.05). A wireless RFA device appears to be useful for measuring implant stability within the limits of the present in vitro study. PMID- 20002530 TI - Surgical and prosthetic reconsiderations in patients with maxillectomy. AB - The purpose of this study was to establish and evaluate new possibilities for rehabilitation of patients with obturator prosthesis who had undergone partial or total maxillectomy because of tumour ablation surgery. Eleven patients with maxillary defects were reconstructed with a computer-aided design/computer-aided manufacturing designed prosthesis. Missing retention was gained by inserting implants in the remaining bone, so that an expansion of the surgical defect to gain further retention could be avoided. All patients were treated successfully according to the previously described treatment plan. The Obturator Functioning Scale (OFS) of the Memorial Sloan-Kettering Cancer Centre was applied to evaluate the functional quality of the obturator prosthesis and patient's satisfaction. It showed good results in all fields of functional outcome and social acceptance. PMID- 20002531 TI - Salivary flow rate and clinical characteristics of patients with xerostomia according to its aetiology. AB - The purpose of the present study was to investigate the differences in salivary flow rates and dry mouth-related subjective symptoms and behaviours in patients with xerostomia according to its aetiology. One hundred and forty patients (24 men and 116 women, mean age, 58.1 +/- 13.3 years) with a chief complaint of xerostomia were included. The patients were divided into five groups; Sjogren's syndrome (n = 34), post-radiation therapy in the head and neck region (n = 16), antipsychotic medications (n = 30), systemic diseases or medications affecting salivary flow (n = 35), and unknown aetiology (n = 25). The patients were asked a standardized series of questions concerning dry mouth, and their whole salivary flow rates were measured. Patients with a history of radiation therapy displayed the most decreased values of salivary flow rates and the most severe associated symptoms and behaviours while patients with unknown aetiology displayed the least decreased values of salivary flow rates and relatively favourable symptoms and behaviours. A burning sensation in the mouth was the most prevalent in patients with systemic diseases or those who were taking medications while altered taste perception was the most prevalent in patients taking antipsychotics. In conclusion, patients with xerostomia displayed various degrees of discomfort related to the quality of life according to the aetiology of their conditions. PMID- 20002532 TI - Classifying degenerative joint disease by the RDC/TMD and by panoramic imaging: a retrospective analysis. AB - The purposes of the study were to evaluate the utility of diagnosing degenerative joint disease (DJD) by the clinical finding of coarse crepitus alone, without supporting imaging studies, as defined by the RDC/TMD, and to evaluate the contribution of panoramic radiography as an aid in the diagnosis of DJD. A retrospective analysis of 372 consecutive patients with TMD was conducted. Their panoramic radiographs were evaluated for the extent of their contribution to the final diagnosis. Panoramic radiography was of no diagnostic value in 94.4% of the cases when the group was considered as a whole. When patients diagnosed with DJD were considered separately, panoramic radiography was completely sufficient for reaching the final diagnosis in 20.0% of the cases. In almost 90% of these patients, however, the clinical examination did not support the diagnosis of DJD (no coarse crepitus was found). This raises some doubts about the effectiveness of the clinical examination according to the RDC/TMD and about the utility of panoramic radiography in the definitive diagnosis of DJD, because both techniques have low accuracy (11.1% and 20%, respectively). The present study supports the current recommendations that panoramic radiography should not be ordered routinely to assess DJD, but still it is first choice when any dental problem is suspected. Further additional imaging (computerized tomography, magnetic resonance imaging) should be considered only if there is reason to expect that the findings might affect diagnosis and management. This study adds to recent criticisms of the clinical validity of the RDC/TMD, with regard to DJD. PMID- 20002533 TI - Oral parafunctions as risk factors for diagnostic TMD subgroups. AB - The frequency of diurnal clenching and/or grinding and nail-biting habits was assessed in patients affected by temporomandibular disorders (TMDs) and in healthy controls in order to investigate the possible association between these oral parafunctions and different diagnostic subgroups of TMDs. The case group included 557 patients (127 men, mean age +/- SD = 34.5 +/- 15.4 years; 430 women, mean age +/- SD = 32.9 +/- 14.1 years) affected by myofascial pain or disc displacement or arthralgia/arthritis/arthrosis. The control group included 111 healthy subjects (55 men, mean age +/- SD = 37 +/- 15.2 years; 56 women, mean age +/- SD = 38.2 +/- 13.8 years). Multinomial logistic regression analysis was used to assess the association between oral parafunctions and TMDs, after adjusting for age and gender. Daytime clenching/grinding was a significant risk factor for myofascial pain (odds ratio (OR) = 4.9, 95% confidence interval (CI): 3.0-7.8) and for disc displacement (OR = 2.5, 95% CI: 1.4-4.3), nail biting was not associated to any of the subgroups investigated. Female gender was a significant risk factor for myofascial pain (OR = 3.8; 95% CI: 2.4-6.1), whereas the risk factor for developing disc displacement decreased with ageing. No association was found between gender, age and arthralgia/arthritis/arthrosis. PMID- 20002534 TI - Relationship between the unilateral TMJ osteoarthritis/osteoarthrosis, mandibular asymmetry and the EMG activity of the masticatory muscles: a retrospective study. AB - The purpose of this retrospective study was to investigate the relationship between the unilateral temporomandibular joint (TMJ) osteoarthritis/osteoarthrosis (OA), mandibular asymmetry and electromyographic (EMG) activity of the masticatory muscles. Twenty-two Japanese women (aged 23.2 +/- 5.4 years) and 10 Japanese men (aged 22.4 +/- 2.8 years) exhibiting unilateral TMJ OA were included in this study. Two angular and seven linear measurements were obtained for the analysis of the skeletal hard tissues. The cephalometric measurement values (CV) were normalized using the CV ratio for the evaluation of the degree of mandibular asymmetry. The EMG was recorded during maximal voluntary clenching efforts for 10 s in the intercuspal position. The average values of integral EMG (iEMG) of three trials were normalized using the iEMG ratio for the evaluation of the functional balance of the masticatory muscles. The mandibular midline was shifted to the TMJ OA side with a median value of 9.85 mm. The CV ratio of the ramus height of the TMJ OA side was significantly smaller than that of the non-OA side. For the masseter muscle, the iEMG ratio of the TMJ OA side was significantly larger than that of the non-OA side (P < 0.05). These results suggest that unilateral TMJ OA is related to the dentofacial morphology, thus resulting in a mandibular midline shift to the affected side and it is associated with a masticatory muscle imbalance. PMID- 20002535 TI - The use of likelihood ratio methodology to find predictors of treatment outcome in patients with dental injury diagnoses. AB - The purpose of this prospective, cohort study of patients with dental injuries was to develop prediction rules to predict treatment response related to the management of dental injuries. The study comprised of 130 patients with a single permanent incisor affected by a dental displacement (n = 100) or fracture injury (n = 30). Laser Doppler flowmetry (LDF) measurements of pulpal blood flow (PBF) were taken 6 and 18 weeks after dental injury Treatment response (success or failure) was categorized based on findings of clinical and radiographical evaluation after 9 months. Forty-four (34%) subjects were categorized as treatment success (absence of loss of sensitivity, periapical radiolucency and grey discolouration of crown), 43 (33%) as treatment failures (loss of sensitivity, and periapical radiolucency and/or grey discolouration of crown) and 43 (33%) as acceptable outcome (loss of sensitivity). After using univariate analysis to determine the association between potential clinical and LDF predictor variables and treatment response status, preliminary prediction rules were developed for prediction of success [positive likelihood ratio (LR), 29.0; 95% confidence interval (CI), 1.7-496.4] and failure (negative LR, 0.55; CI, 0.4 0.7). The most important variables were subluxation, root fracture, baseline PBF level and change in PBF level at 3-month follow-up. Outcome following the management of dental injuries may be predicted from variables collected from LDF and physical examination. Predictive modelling may provide clinicians with the opportunity to identify 'at-risk' patients early and initiate specific treatment approaches. PMID- 20002536 TI - The future of complete dentures in oral rehabilitation. A critical review. AB - Based on available investigations and current trends in oral rehabilitation published in the dental literature, an attempt is made to describe the possible future role of complete dentures. For edentulous patients, complete dentures have for long been the only prosthodontic treatment option. Whereas a large number of edentulous patients report satisfaction with denture usage, a smaller number are unable to adapt; for such patients, sophistication of clinical and technical processes or quality of denture-supporting tissues, appear to have little influence on patient-perceived outcomes. Since the 1980s, osseointegrated dental implants have dramatically improved the therapeutic possibilities, especially so for maladaptive patients. Those able to access such treatment can expect significant improvements in oral functional status and quality of life. While there is a downward trend in edentulism in several countries, it is region specific, confirming the overriding influence of socio-economic factors on health status. In most societies, despite ageing populations, the need for complete dentures is not likely to reduce in the near future. Whereas a two- or even a one implant overdenture for the edentulous mandible is increasingly regarded as a minimum standard of care in many developed countries, its routine prescription for the majority in the world who are disadvantaged is unrealistic; for them, even 'low-tech' therapies like conventional dentures are beyond their reach. Improving the conventional management of edentulous patients is a necessity and requires a keener focus by researchers, educators and clinicians in the developed world on the needs of populations with fewer resources. PMID- 20002537 TI - Which device is more accurate to determine the stability/mobility of dental implants? A human cadaver study. AB - Non-invasive devices including resonance frequency (RF) analysis and mobility measuring (MM) damping capacity assessment are used to measure implant stability/mobility. The aims of the study were to compare the primary stability of implant inserted into extraction sockets by using RF with cable, RF wireless and new wireless MM device, to clarify the relation between these devices and to understand the correlations between peri-implant bone levels and implant stability. A total of 30 screw-type implants (3.75 x 11 and 4.2 x 11 mm) were inserted into extraction sockets of eight mandibular pre-molar regions of human cadavers. The primary stability of implants was measured by three devices after insertion. Peri-implant vertical defects were created in millimetre increments ranging between 0 and 5 mm, and stability/mobility of implants were analysed. At placement, the mean implant stability quotient of RF with cable, RF wireless and MM device values was 46 +/- 1, 57.8 +/- 9 and -5.4 +/- 1, respectively. Statistical correlations were demonstrated between these devices (P = 0.001). Statistically significant differences were presented for all peri-implant detects ranging between 0 and 5 mm for RF with cable and RF wireless at all increments. However, only a significant decrease was found between 0 and 1 mm defects, and 4 and 5 mm defects in MM device. Although RF with cable and RF wireless seem to be suitable to detect peri-implant bone loss around implants in 1 mm increments, the new MM device may not be suitable to detect the 1 mm peri-implant bone changes in human dried cadaver mandibles. PMID- 20002538 TI - Hereditary and acquired protein S deficiencies are associated with low TFPI levels in plasma. AB - BACKGROUND: Protein S and tissue factor pathway inhibitor (TFPI) act together in down-regulating coagulation. OBJECTIVE: To investigate the TFPI/protein S system in hereditary and acquired protein S deficiency. METHODS: Plasma antigen levels of protein S and full-length TFPI were determined in heterozygous type I protein S-deficient individuals (n=35), patients on oral anticoagulant treatment (OAT) (n=29), oral contraceptive (OC) users (n=10) and matched controls. Thrombin generation was determined using calibrated automated thrombography. RESULTS: Full length TFPI levels were lower in type I protein S-deficient individuals (76.8+/ 33.8%) than in age- and sex-matched controls (128.0+/-59.4%, P<0.001). Among protein S-deficient individuals with thrombosis, those on OAT had not only lower total protein S levels (25.7+/-8.2% vs. 54.7+/-8.2%, P<0.001), but also lower full-length TFPI levels (52.6+/-15.0% vs. 75.4+/-22.9%, P=0.009) than those not on OAT. Similarly, OC users had lower protein S (73.8+/-11.5% vs. 87.9+/-10.8%, P=0.005) and full-length TFPI levels (73.7+/-27.7% vs. 106.4+/-29.2%, P=0.007) than non-users. When triggered with tissue factor, plasma from protein S deficient individuals generated 3-5-fold more thrombin than control plasma. The difference was only partially corrected by normalization of the protein S level, full correction requiring additional normalization of the TFPI level. Protein S immunodepletion experiments indicated that free protein S and full-length TFPI form a complex in plasma, and the protein S/TFPI interaction was confirmed by surface plasmon resonance analysis. CONCLUSIONS: Full-length TFPI binds to protein S in plasma and is reduced in genetic and acquired protein S deficiency. The concomitant TFPI deficiency substantially contributes to the hypercoagulable state associated with protein S deficiency. PMID- 20002539 TI - APC resistance: biological basis and acquired influences. AB - Proteolytic inactivation of factors Va (FVa) and VIIIa (FVIIIa) by activated protein C (APC) and its cofactors protein S and factor V (FV) is a key process in the physiological down-regulation of blood coagulation. Functional abnormalities of this pathway, which manifest themselves in vitro as a poor anticoagulant response of plasma to added APC (APC resistance), are prevalent in the general population and are associated with an increased risk of venous thrombosis. APC resistance was originally discovered in thrombophilic families and later shown to be associated with the common FV Arg506Gln (FV(Leiden)) mutation, which abolishes one of the APC-cleavage sites in FV. Although FV(Leiden) is the major cause of hereditary APC resistance, it is becoming increasingly clear that several other genetic and acquired conditions contribute to APC resistance and thereby increase the risk of venous thrombosis. This paper reviews the multifactorial etiology of APC resistance and discusses its clinical implications. PMID- 20002540 TI - Increased sperm count may account for high population frequency of factor V Leiden. AB - BACKGROUND: Factor V Leiden (FVL) increases the risk of venous thrombosis and pregnancy loss in carriers. Nevertheless, this relatively old mutation is prevalent in Caucasion populations, which could be explained by positive selection pressure. Men with FVL have previously been found to have higher fecundity (the time between marriage and first pregnancy). Whether this is caused by increased sperm counts in men with FVL is unknown. OBJECTIVES: To assess whether men with factor V Leiden have increased sperm counts. PATIENTS/METHODS: We performed a prospective cohort study among 1139 consecutively included male partners of subfertile couples presenting at our university hospital for fertility workup between January 2000 and July 2007. All potential candidates who gave informed consent were included, irrespective of their fertility workup. In this retrospective analysis, we excluded participants with known causes of spermatogenic function or azoospermia. Subsequently, we genotyped all participants and compared sperm counts between FVL carriers and non-carriers. RESULTS: We identified 37 FVL carriers and 921 non-carriers. FVL carriers had higher total sperm counts and total motile sperm counts than non-carriers: 236 x 10(6) (95% CI 158-292 x 10(6)) vs. 163 x 10(6) (95% CI 147-178 x 10(6)) and 81 x 10(6) (95% CI 54-105 x 10(6)) vs. 52 x 10(6) (95% CI 48-57 x 10(6)), respectively. CONCLUSIONS: To our knowledge, this is the first study that indicates that an increased incidence of a genotype may be controlled by increased sperm counts. However, the finding that men with FVL had higher total (motile) sperm counts was not statistically significant and needs confirmation in other studies. PMID- 20002541 TI - Mlck1a is expressed in zebrafish thrombocytes and is an essential component of thrombus formation. AB - BACKGROUND: We have used the advantages of the zebrafish model system to demonstrate which of the vertebrate myosin light chain kinase (MLCK) genes is expressed in thrombocytes and important for thrombus formation. METHODS AND RESULTS: Here we report that Mlck1a is an essential component of thrombus formation. Phylogenetic data revealed four zebrafish orthologous for three human MLCK genes. To investigate expression of the zebrafish mlck genes in thrombocytes we compared GFP-tagged platelets with other cells by microarray analysis, and showed that mlck1a expression was 4.5-fold enriched in platelets. Furthermore, mlck1a mRNA and mRNA for the platelet-specific cd41 co-localized in thrombi. Expression of other mlck subtypes was lower in GFP-tagged platelets (mlck1b; 0.77 fold enriched) and absent in thrombi (mlck1b, -2, -3). To investigate the role of Mlck1a in thrombus formation, we knocked down mlck1a using two morpholinos. This resulted in impaired morphology changes of platelets adhering on fibrinogen. In a thrombosis model, in which thrombocytes adhere to the vessel wall damaged by laser irradiation, thrombus formation was slowed down in mlck1a-deficient embryos. CONCLUSION: We conclude that Mlck1a is the subtype of MLCK that contributes to platelet shape change and thrombus formation. PMID- 20002542 TI - Apolipoprotein C-III predicts cardiovascular mortality in severe coronary artery disease and is associated with an enhanced plasma thrombin generation. AB - BACKGROUND: Apolipopoprotein C-III (apo C-III) plays a pivotal role in controlling plasma triglyceride (TG) and contributes to the atherogenic properties of TG-rich lipoproteins. OBJECTIVES: (i) To examine the predictive value of serum apo C-III for cardiovascular mortality in the setting of secondary prevention of coronary artery disease (CAD); and (ii) to evaluate possible associations between apolipoprotein levels and the thrombin generation assay, a global test to estimate plasma thrombogenic potential. METHODS AND RESULTS: A cohort of 633 patients with angiographically proven CAD was prospectively followed for a median follow-up of 57 months. The large majority of them (92%) underwent coronary (endovascular or surgical) revascularization. During the follow-up, 91 (14.3%) out of 633 patients died, with 64 events (10.1%) attributed to cardiovascular causes. After adjustment for all the other predictors of mortality during univariate analysis (i.e. age, statin therapy, myocardial infarction history, diabetes, hs-CRP and creatinine), elevated apo C-III levels (> or = 10.5 mg dL(-1)- the median value) significantly predicted both total and cardiovascular mortality (HR for total mortality 2.22 with 95% CI 1.16-4.24; HR for cardiovascular mortality 2.35 with 95% CI 1.19-4.62). In a subgroup of 225 subjects, apo C-III levels were significantly associated with endogenous thrombin potential in regression models (standardized beta coefficient = 0.207, P = 0.002). CONCLUSIONS: Basal concentrations of apo C-III levels > or = 10.5 mg dL( 1) in CAD patients independently predicted cardiovascular mortality during the subsequent 5-year period. Such concentrations were associated with an enhanced plasma endogenous thrombin generation, suggesting a complex interplay between TG rich particles and the coagulation cascade as well as a new 'thrombogenetic' role for apo C-III. PMID- 20002543 TI - RGD-ligand mimetic antagonists of integrin alphaIIbbeta3 paradoxically enhance GPVI-induced human platelet activation. AB - BACKGROUND: The integrin alpha(IIb)beta(3) is the major mediator of platelet aggregation and has, therefore, become an important target of antithrombotic therapy. Antagonists of alpha(IIb)beta(3), for example abciximab, tirofiban and eptifibatide, are used in the treatment of acute coronary syndromes. However, in addition to effective blockade of the integrin, binding of can induce conformational changes in the integrin and can also induce integrin clustering. This class effect of RGD-ligand mimetics might, therefore, underlie paradoxical platelet activation and thrombosis previously reported. OBJECTIVES: To examine the components of signaling pathways and functional responses in platelets that may underlie this phenomenon of paradoxical platelet activation. METHODS: We assessed the effect of lotrafiban, and other alpha(IIb)beta(3) antagonists including the clinically used drug tirofiban, on tyrosine phosphorylation of key signaling proteins in platelets by immunoblotting and also platelet functional outputs such as cytosolic calcium responses, phosphatidylserine exposure (pro coagulant activity) and dense granule release. RESULTS: In all cases, no effect of alpha(IIb)beta(3) antagonists were observed on their own, but these integrin antagonists did lead to a marked potentiation of glycoprotein VI (GPVI) associated FcR gamma-chain phosphorylation, activation of Src family kinases and Syk kinase. This correlated with increased dense granule secretion, cytosolic calcium response and exposure of phosphatidylserine on the platelet surface. P2Y(12) antagonism abolished the potentiated phosphatidylserine exposure and dense granule secretion but not the cytosolic calcium response. CONCLUSIONS: These data provide a mechanism for enhancement of platelet activity by alpha(IIb)beta(3) inhibitors, but also reveal a potentially important signaling pathway operating from the integrin to GPVI signaling. PMID- 20002544 TI - Polyphosphate binds with high affinity to exosite II of thrombin. AB - BACKGROUND: Polyphosphate (a linear polymer of inorganic phosphate) is secreted from platelet dense granules, and we recently showed that it accelerates factor V activation by thrombin. OBJECTIVE: To examine the interaction of polyphosphate with thrombin. METHODS AND RESULTS: Thrombin, but not prothrombin, altered the electrophoretic migration of polyphosphate in gel mobility assays. Thrombin binding to polyphosphate was influenced by ionic strength, and was evident even in plasma. Two positively charged exosites on thrombin mediate its interactions with other proteins and accessory molecules: exosite I (mainly with thrombin substrates), and exosite II (mainly with certain anionic polymers). Free thrombin, thrombin in complex with hirudin's C-terminal dodecapeptide and gamma thrombin all bound polyphosphate similarly, excluding exosite I involvement. Mutations within exosite II, but not within exosite I, the Na(+)-binding site or hydrophobic pocket, weakened thrombin binding to polyphosphate as revealed by NaCl dependence. Surface plasmon resonance demonstrated tight interaction of polyphosphate with thrombin (K(d) approximately 5 nm) but reduced interaction with a thrombin exosite II mutant. Certain glycosaminoglycans, including heparin, only partially competed with polyphosphate for binding to thrombin, and polyphosphate did not reduce heparin-catalyzed inactivation of thrombin by antithrombin. CONCLUSION: Polyphosphate interacts with thrombin's exosite II at a site that partially overlaps with, but is not identical to, the heparin-binding site. Polyphosphate interactions with thrombin may be physiologically relevant, as the polyphosphate concentrations achievable following platelet activation are far above the approximately 5 nM K(d) for the polyphosphate-thrombin interaction. PMID- 20002545 TI - Diverse functions of protein kinase C isoforms in platelet activation and thrombus formation. AB - Platelet activation is a complex balance of positive and negative signaling pathways. The protein kinase C (PKC) family is a major regulator of platelet granule secretion, integrin activation, aggregation, spreading and procoagulant activity. As broad-spectrum PKC inhibitors reduce secretion and aggregation, the PKC family is generally considered to be a positive regulator of platelet activation. However, the individual members of the PKC family that are expressed in platelets are regulated in different ways, and an increasing body of evidence indicates that they have distinct, and often opposing, roles. Many of the recent advances in understanding the contributions of individual PKC isoforms have come from mouse gene knockout studies. PKCalpha, a classic isoform, is an essential positive regulator of granule secretion and thrombus formation, both in vitro and in vivo. Mice lacking PKCalpha show much reduced thrombus formation in vivo but do not have a bleeding defect, suggesting that PKCalpha could be an attractive antithrombotic target. Important, apparently non-redundant, roles, both positive and negative, for the novel PKC isoforms delta, theta and epsilon in granule secretion have also been proposed, indicating highly complex regulation of this essential process. Similarly, PKCbeta, PKCdelta and PKCtheta have non-redundant roles in platelet spreading, as absence of either PKCbeta or PKCtheta reduces spreading, whereas PKCdelta negatively regulates filopodial formation. This negative signaling by PKCdelta may reduce platelet aggregation and so restrict thrombus formation. In this review, we discuss the current understanding of the regulation and functions of individual PKC isoforms in platelet activation and thrombus formation. PMID- 20002546 TI - The REFLECT statement: methods and processes of creating reporting guidelines for randomized controlled trials for livestock and food safety. AB - The conduct of randomized controlled trials in livestock with production, health, and food-safety outcomes presents unique challenges that might not be adequately reported in trial reports. The objective of this project was to modify the CONSORT (Consolidated Standards of Reporting Trials) statement to reflect the unique aspects of reporting these livestock trials. A 2-day consensus meeting was held on November 18-19, 2008 in Chicago, IL, to achieve the objective. Before the meeting, a Web-based survey was conducted to identify issues for discussion. The 24 attendees were biostatisticians, epidemiologists, food-safety researchers, livestock production specialists, journal editors, assistant editors, and associate editors. Before the meeting, the attendees completed a Web-based survey indicating which CONSORT statement items would need to be modified to address unique issues for livestock trials. The consensus meeting resulted in the production of the REFLECT (Reporting Guidelines for Randomized Control Trials) statement for livestock and food safety and 22-item checklist. Fourteen items were modified from the CONSORT checklist, and an additional subitem was proposed to address challenge trials. The REFLECT statement proposes new terminology, more consistent with common usage in livestock production, to describe study subjects. Evidence was not always available to support modification to or inclusion of an item. The use of the REFLECT statement, which addresses issues unique to livestock trials, should improve the quality of reporting and design for trials reporting production, health, and food-safety outcomes. PMID- 20002547 TI - Babesiosis caused by a large Babesia species in 7 immunocompromised dogs. AB - BACKGROUND: A large unnamed Babesia species was detected in a dog with lymphoma. It was unknown if this was an underrecognized pathogen. OBJECTIVE: Report the historical and clinicopathologic findings in 7 dogs with babesiosis caused by a large unnamed Babesia species characterize the 18S ribosomal ribonucleic acid (rRNA) genes. ANIMALS: Seven immunocompromised dogs from which the Babesia was isolated. METHODS: Retrospective case review. Cases were identified by a diagnostic laboratory, the attending clinicians were contacted and the medical records were reviewed. The Babesia sp. 18S rRNA genes were amplified and sequenced. RESULTS: Six of 7 dogs had been splenectomized; the remaining dog was receiving oncolytic drugs. Lethargy, anorexia, fever, and pigmenturia were reported in 6/7, 6/7, 4/7, and 3/7 dogs. Laboratory findings included mild anemia (7/7) and severe thrombocytopenia (6/7). Polymerase chain reaction (PCR) assays used to detect Babesia sensu stricto species were all positive, but specific PCR assays for Babesia canis and Babesia gibsoni were negative in all dogs. The 18S rRNA gene sequences were determined to be identical to a large unnamed Babesia sp. previously isolated. Cross-reactive antibodies against other Babesia spp. were not always detectable. Five dogs were treated with imidocarb dipropionate and 1 dog with atovaquone/azithromycin; some favorable responses were noted. The remaining dog was untreated and remained a clinically stable carrier. CONCLUSIONS AND CLINICAL IMPORTANCE: Dogs with pigmenturia, anemia, and thrombocytopenia should be tested for Babesia sp. by PCR. Serology is not sufficient for diagnosis of this Babesia sp. Asplenia, chemotherapy, or both might represent risk factors for persistent infection, illness, or both. PMID- 20002548 TI - Antemortem diagnosis of a distal axonopathy causing severe stringhalt in a horse. PMID- 20002549 TI - Von Willebrand factor antigen concentration in dogs with sepsis. AB - BACKGROUND: Von Willebrand factor (vWF) antigen concentration, a marker of endothelial activation, is increased in human patients with multiorgan failure, sepsis, or both, and is an independent predictor of survival. HYPOTHESIS/OBJECTIVES: vWF antigen concentrations are significantly higher in dogs with sepsis. ANIMALS: Fourteen dogs hospitalized with sepsis. Sepsis was defined as microbiologic or cytologic evidence of infection combined with systemic inflammatory response syndrome. Control dogs were healthy dogs, without evidence of disease. METHODS: Prospective, observational study. Dogs admitted to the intensive care unit with a diagnosis of sepsis were considered eligible for enrollment into the study. Exclusion criteria included a previous diagnosis of von Willebrand disease or a recent history of a plasma transfusion. Citrated plasma samples were collected for analysis of vWF antigen by ELISA. All samples were drawn from dogs during hospitalization. Data between populations were analyzed using nonparametric statistical analysis with a P value < .05 considered significant. RESULTS: Twenty-five dogs were enrolled; 14 dogs with sepsis and 11 control dogs. The median vWF antigen concentration in dogs with sepsis was 156% (range, 117-200%), which was significantly higher than healthy dogs (105%; range, 44-155%, P < .005). There was no difference between survivors and nonsurvivors with a median vWF antigen concentration of 144% (range, 136-201%) in survivors (n = 7) and 159% (range, 122-174%) in nonsurvivors (n = 7) (P = .5). CONCLUSIONS AND CLINICAL IMPORTANCE: vWF is increased in dogs with sepsis, possibly reflecting endothelial activation. Further exploration of endothelial function is warranted in critically ill dogs. PMID- 20002550 TI - Quality matters: publishing in the era of CONSORT, REFLECT, and EBM. PMID- 20002551 TI - Effect of tissue processing on assessment of endoscopic intestinal biopsies in dogs and cats. AB - BACKGROUND: Prior studies failed to detect significant association between hypoalbuminemia and small intestinal lesions. HYPOTHESIS: Use of pictorial templates will enhance consistency of interpathologist interpretation and identification of intestinal lesions associated with hypoalbuminemia. ANIMALS: Tissues from 62 dogs and 25 cats examined as clinical cases at 7 referral veterinary practices in 4 countries. METHODS: Retrospective, observational study. Histopathology slides from sequential cases undergoing endoscopic biopsy were examined by 4 pathologists by pictorial templates. Changes for 9 microscopic features were recorded as normal, mild, moderate or severe, and 2- and 4-point scales were tested for consistency of interpretation. Logistic regression models determined odds ratios (OR) of histologic lesions being associated with hypoalbuminemia while kappa statistics determined agreement between pathologists on histologic lesions. RESULTS: There was poor agreement (kappa = -0.013 to 0.3) between pathologists, and institution of origin of slides had effect (kappa = 1.0 for 3 of 4 lesions on slides from Institution 5) on agreement between pathologists on selected histologic features. Using 2 point as opposed to 4-point grading scale increased agreement between pathologists (maximum kappa = 0.69 using 4-point scale versus maximum kappa = 1.0 using 2-point scale). Significant association (P = .019- .04; 95% OR = 3.14-10.84) between lacteal dilation and hypoalbuminemia was found by 3 pathologists. CONCLUSIONS AND CLINICAL IMPORTANCE: Substantial inconsistency between pathologists remains despite use of pictorial template because of differences in slide processing. Distinguishing between mild and moderate lesions might be important source of the disagreement among pathologists. PMID- 20002552 TI - Serum cardiac troponin I concentration in dogs with precapillary and postcapillary pulmonary hypertension. AB - BACKGROUND: Pulmonary hypertension (PH) is a disease condition leading to right sided cardiac hypertrophy and, eventually, right-sided heart failure. Cardiac troponin I (cTnI) is a circulating biomarker of cardiac damage. HYPOTHESIS: Myocardial damage can occur in dogs with precapillary and postcapillary PH. ANIMALS: One hundred and thirty-three dogs were examined: 26 healthy controls, 42 dogs with mitral valve disease (MVD) without PH, 48 dogs with pulmonary hypertension associated with mitral valve disease (PH-MVD), and 17 dogs with precapillary PH. METHODS: Prospective, observational study. Serum cTnI concentration was measured with a commercially available immunoassay and results were compared between groups. RESULTS: Median cTnI was 0.10 ng/mL (range 0.10 0.17 ng/mL) in healthy dogs. Compared with the healthy population, median serum cTnI concentration was increased in dogs with precapillary PH (0.25 ng/mL; range 0.10-1.9 ng/mL; P < .001) and in dogs with PH-MVD (0.21 ng/mL; range 0.10-2.10 ng/mL; P < .001). Median serum cTnI concentration of dogs with MVD (0.12 ng/mL; range 0.10-1.00 ng/mL) was not significantly different compared with control group and dogs with PH-MVD. In dogs with MVD and PH-MVD, only the subgroup with decompensated PH-MVD had significantly higher cTnI concentration compared with dogs with compensated MVD and PH-MVD. Serum cTnI concentration showed significant modest positive correlations with the calculated pulmonary artery systolic pressure in dogs with PH and some echocardiographic indices in dogs with MVD and PH-MVD. CONCLUSIONS AND CLINICAL IMPORTANCE: Serum cTnI is high in dogs with either precapillary and postcapillary PH. Myocardial damage in dogs with postcapillary PH is likely the consequence of increased severity of MVD. PMID- 20002553 TI - Clinical features and outcome of Heterobilharzia americana infection in dogs. AB - BACKGROUND: Heterobilharzia americana (HA), the causative agent of canine schistosomiasis, is a flatworm with a freshwater snail as an intermediate host. Only case reports or small case series evaluating naturally infected dogs have been published. OBJECTIVE: Describe clinical signs in dogs naturally infected with HA. ANIMALS: Twenty-two dogs naturally infected with HA from 1985 to 2009. METHODS: Retrospective study. All medical records were searched for HA and schistosomiasis. Only dogs with a diagnosis based on identification of ova on histopathology or fecal saline sedimentation were included. RESULTS: The median age was 3.1 years (1-12). The median duration of clinical signs before diagnosis was 0.63 months (0.03-12). The most common clinical signs were lethargy (91%), weight loss (77%), hyporexia (68%), vomiting (59%), and diarrhea (55%). Eleven of the 22 dogs were hypercalcemic. Hypercalcemia did not resolve without definitive treatment with praziquantel. HA infection was an incidental diagnosis in 7/22 dogs. Diagnosis was obtained via necropsy (4), histopathology (9), and fecal examination (9). Definitive treatment included praziquantel and fenbendazole. Eighteen dogs were diagnosed antemortem and 17 were treated. Twelve dogs were alive for 6 months to 3 years after diagnosis. CONCLUSIONS AND CLINICAL IMPORTANCE: HA infection occurs in younger, larger breed, indoor dogs. Hypercalcemia does not resolve without praziquantel treatment. Prognosis is good and neither hypercalcemic-induced renal failure nor ascites appears to worsen prognosis. Dogs in affected areas or that have traveled to affected areas that present for weight loss, gastrointestinal or liver disease, and hypercalcemia, should be tested. PMID- 20002554 TI - Cardiac troponin I is associated with severity of myxomatous mitral valve disease, age, and C-reactive protein in dogs. AB - BACKGROUND: Concentrations of cardiac troponin I (cTnI) and C-reactive protein (CRP) might be associated with cardiac remodeling in dogs with myxomatous mitral valve disease (MMVD). Age- and sex-dependent variations in cTnI concentration have been described. OBJECTIVE: To investigate whether plasma concentrations of cTnI and CRP are associated with severity of MMVD, and investigate potential associations of dog characteristics on cTnI and CRP concentrations. ANIMALS: Eighty-one client-owned dogs with MMVD of varying severity. METHODS: Dogs were prospectively recruited for the study. Dogs were classified according to severity of MMVD. Plasma cTnI was analyzed by a high sensitivity cTnI assay with a lower limit of detection of 0.001 ng/mL, and plasma CRP was analyzed by a canine specific CRP ELISA. RESULTS: Higher cTnI concentrations were detected in dogs with moderate (0.014 [interquartile range 0.008-0.029] ng/mL, P = .0011) and severe (0.043 [0.031-0.087] ng/mL, P < .0001) MMVD, compared with healthy dogs (0.001 [0.001-0.004]ng/mL). Dogs with severe MMVD also had higher cTnI concentrations than dogs with mild (0.003 [0.001-0.024] ng/mL, P < .0001) and moderate (P = .0019) MMVD. There were significant associations of age, CRP, heart rate, and left ventricular end-diastolic diameter, on cTnI concentration C reactive protein did not differ among severity groups, but was significantly associated with cTnI, breed, and systolic blood pressure on CRP concentration. CONCLUSIONS AND CLINICAL IMPORTANCE: Analysis of cTnI concentration has potential to increase knowledge of overall cardiac remodeling in dogs with MMVD. However, effect of age on cTnI needs consideration when assessing cTnI. PMID- 20002555 TI - Comparison of histopathologic findings in biopsies from the duodenum and ileum of dogs with enteropathy. AB - BACKGROUND: In the investigations of dogs with chronic small intestinal diarrhea collection of ileal biopsies lengthens procedural time and has been of uncertain value. OBJECTIVES: To evaluate whether there was agreement between histologic changes present in samples of duodenal and ileal mucosa, and hence to provide initial information in the process of determining whether collection of ileal biopsies is clinically justified. ANIMALS: 40 dogs with chronic small and large intestinal diarrhea from which endoscopic (in 30 cases) or surgical (in 10 cases) duodenal and ileal biopsies had been collected. METHODS: Samples were reviewed concurrently by two observers (MJD and MDW) using the scoring system developed by the World Small Animal Veterinary Association (WSAVA) Gastrointestinal Standardization Group. Comparisons were made by kappa analysis. RESULTS: Microscopic pathology was observed in 30 cases. Only eight out of this 30 (27%) had the same histopathologic diagnosis in both the duodenum and the ileum. This dropped to 3 out of 30 (10%) if different disease severity was also considered as disagreement. Microscopic pathology would have been found in 60% and 80% of the 30 cases, if only duodenal or ileal biopsies respectively, had been available. CONCLUSIONS AND CLINICAL IMPORTANCE: There was poor agreement between histopathological findings from duodenal versus ileal biopsies with abnormalities sometimes being more readily detected in the ileum. Routine collection of ileal plus duodenal samples appears warranted when concurrent small and large intestinal diarrhea is present. PMID- 20002556 TI - Evaluation of a novel real-time continuous glucose-monitoring system for use in cats. AB - BACKGROUND: The Guardian REAL-Time is a continuous glucose-monitoring system (CGMS) recently developed to provide instantaneous interstitial glucose concentrations; the system does not require a monitor being fixed to the animal. HYPOTHESIS: The CGMS provides accurate and reproducible real-time readings of glucose concentration in cats. ANIMALS: Thirty-two diabetic cats, 2 cats with suspected insulinoma, and 5 healthy cats. METHODS: Prospective, observational study. CGMS accuracy was compared with a reference glucose meter at normal, high, and low blood glucose concentrations using error grid analysis. Reading variability of 2 simultaneously used CGMS was determined in diabetic cats by calculating correlation and percentage of concordance of paired data at different glycemic ranges. The time interval between increasing glycemia and a rise in interstitial fluid glucose measured by the CGMS was assessed in healthy cats receiving glucose IV; the time point of maximal increase in interstitial glucose concentrations was calculated. RESULTS: The CGMS was 100, 96.1, and 91.0% accurate at normal, high, and low blood glucose concentrations. Measurements deviated from reference by -12.7 +/- 70.5 mg/dL at normal, -12.1 +/- 141.5 mg/dL at high, and -1.9 +/- 40.9 mg/dL at low glucose concentrations. Overall, paired CGMS readings correlated significantly (r = 0.95, P < .0001) and concordance was 95.7%. The median delay after IV administration of glucose to an increase in interstitial glucose was 11.4 minutes (range: 8.8-19.7 minutes). CONCLUSIONS AND CLINICAL IMPORTANCE: Although some readings substantially deviated from reference values, the CGMS yields reproducible results, is clinically accurate in cats with hyperglycemia and euglycemia, and is slightly less accurate if blood glucose concentrations are low. Rapidly increasing interstitial glucose after a glycemic rise suggests that the CGMS is suitable for real-time measurement under clinical conditions. PMID- 20002557 TI - Prevalence of Staphylococcus aureus and methicillin-resistant Staphylococcus aureus carriage in three populations. AB - BACKGROUND: A higher prevalence of methicillin-resistant Staphylococcus aureus (MRSA) colonization is reported in healthcare workers compared with nonhealthcare workers. HYPOTHESIS: The prevalence of MRSA colonization differed in people and pets in households with healthcare workers as compared with households without healthcare workers. SUBJECTS: A person and 1 dog or cat from 586 households defined as either a nonhealthcare (n = 213), veterinary healthcare (n = 211), or human healthcare (n = 162) worker household. METHODS: Prospective cross-sectional study. Samples from humans and pets were cultured in vitro. Staphylococcus aureus was identified as methicillin sensitive (MSSA) or MRSA with mecA polymerase chain reaction. Pulsed-field gel electrophoresis and spa-typing were used to characterize relatedness of S. aureus and MRSA and assign USA types. RESULTS: The prevalence of MSSA and MRSA in humans was 21.5% (126/586) and 5.63% (33/586), respectively, and 7.85% (46/586) and 3.41% (20/586), respectively, in pets. There were no differences in prevalences of either MSSA or MRSA between household types. The proportion of MRSA among all S. aureus isolates in humans and pets was 20.8% (33/159) and 30.3% (20/66), respectively. In < 1.0% (4/586) of households, the same strain of MRSA was found in both a person and a pet. CONCLUSIONS AND CLINICAL IMPORTANCE: There were no differences in the prevalences of MSSA or MRSA between healthcare worker and nonhealthcare worker households. Pets and people colonized with S. aureus were as likely to be colonized with MRSA. Colonization of a person and their pet with the same strain of MRSA was rare. PMID- 20002558 TI - Hepatitis C/HIV co-infection is associated with higher mortality in hospitalized patients with hepatitis C or HIV. AB - Up to 10% of all patients with Hepatitis C virus (HCV) infection are co-infected with human immunodeficiency virus (HIV); 25-30% of HIV patients are co-infected with HCV. The aim of this study was to examine the association of HCV/HIV co infection with outcomes of hospitalized patients compared to those with HCV or HIV monoinfection. Using the 2006 Nationwide Inpatient Sample, patients with HCV or HIV monoinfection or HCV/HIV co-infection were identified using ICD-9-CM codes. We compared liver-related and infection-related admission between the three groups of patients. Multivariate logistic regression was performed to identify independent predictors of in-hospital mortality. A total of 474,843 discharges with HCV monoinfection, 206,758 with HIV monoinfection and 56,304 with HCV/HIV co-infection were included. Liver-related admissions were more common in co-infected patients (15.4%) compared to those with HIV monoinfection (3.3%, P < 0.001). Primary infectious hospitalizations were more common in HIV monoinfection (33.9%) compared to co-infected patients (26%, P < 0.001). HCV/HIV co-infection was associated with higher mortality compared to HCV monoinfection (OR 1.41, 95% CI 1.20-1.65) but not when compared to monoinfected-HIV patients. HCV-associated cirrhosis or complications thereof conferred four times greater mortality risk in patients with HIV (OR 3.96, 95% CI 3.29-4.79). The rate of hospitalization for HCV/HIV co-infected patients (23.5%) was significantly higher than those with HCV (14.8%) or HIV (19.9%) (P < 0.001). HCV/HIV co-infection is associated with significantly higher rates of hospitalization and is a risk factor for in hospital mortality compared to patients with isolated HCV. PMID- 20002559 TI - Health care risk factors among women and personal behaviours among men explain the high prevalence of hepatitis C virus infection in Karachi, Pakistan. AB - To estimate the prevalence and identify factors associated with hepatitis C virus (HCV) infection among men and women in Karachi, Pakistan. We conducted a cross sectional study of adult men and women in a peri-urban community of Karachi (Jam Kandah). Households were selected through systematic sampling from within all villages in the study area. All available adults within each household were interviewed about potential HCV risk factors. A blood specimen was collected to test for anti-HCV antibodies by enzyme immunoassay. We used generalized estimating equations while accounting for correlation of responses within villages to identify the factors associated with HCV infection. Of 1997 participants, 476 (23.8%) were anti-HCV positive. Overall, HCV infection was significantly associated with increasing age, ethnicity, and having received > or =2 blood transfusions, > or =3 hospitalizations, dental treatment and >5 injections among women. Among women, > or =2 blood transfusions [adjusted odds ratio (aOR) = 2.32], >5 injections during the past 6 months (aORs = 1.47), dental treatment (aOR = 1.31) and increasing age(aOR = 1.49), while among men, extramarital sexual intercourse (aOR = 2.77), at least once a week shave from barber (aOR = 5.04), > or =3 hospitalizations (aOR = 2.50) and increasing age (aOR = 1.28) were associated with HCV infection. A very high prevalence of HCV was found in the study population. Among women, unsafe health care practices, while among men extramarital sex, shaving from a barber and hospitalizations were associated with HCV infection. Efforts are needed to improve the safety of medical procedures to reduce the transmission of HCV in Pakistan [Corrections made in Summary after initial online publication.]. PMID- 20002561 TI - The impact of a managed care network on attendance, follow-up and treatment at a hepatitis C specialist centre. AB - Infection with the hepatitis C virus commonly occurs in patient groups who have difficulty accessing conventional medical care, reducing their chance of successful antiviral therapy. Managed care networks (MCNs) have been suggested as a mechanism of improving access to care; however, there is little evidence to support their use in patients with hepatitis C. The aim of this study was to evaluate the impact of a MCN for patients with hepatitis C. This was a retrospective cohort study of all individuals in our area who had received a positive hepatitis C antibody test between August 1994 and June 2008. The MCN introduced a new referral pathway, which included nonmedical referrals and outreach nurse-led clinics. These interventions were introduced in 2004 and evaluated in 2008. After the introduction of the MCN, the proportion of individuals who accessed care increased from 61% (280/430) to 82.4% (721/875). There was an increase in nonmedical referrals with 81 (18.3%) being directly referred from Drug Problem Services and 75 (17%) from the Prison Service. The changes to referral did not have a negative impact on treatment outcomes as the number who completed treatment increased from 66.1% (43/65) to 73.7% (98/133) and the sustained virological response increased from 50.7% (33/65) to 60.9% (81/133). This study provides evidence that the collaboration of health care professionals within a network can have a radical effect in improving access to care in a traditionally hard to reach population. This has been achieved with little additional resource, but rather working smarter with existing staff. PMID- 20002560 TI - Prevalence and factors associated with significant liver fibrosis assessed by transient elastometry in HIV/hepatitis C virus-coinfected patients. AB - Transient elastometry (TE) could provide a more accurate evaluation of the frequency and risk factors of liver fibrosis in hepatitis C virus (HCV) infection than that based on biopsy. The aim of this study was to assess the prevalence of and factors associated with significant liver fibrosis in a large population of HIV/HCV-coinfected patients. HIV/HCV-coinfected patients, who had participated in a cross-sectional, multicenter, retrospective study of liver fibrosis using noninvasive markers and in whom a determination of liver stiffness (LS) by TE was available, were included in this analysis. Factors potentially associated with significant fibrosis (LS >= 9 kPa) were analyzed. One thousand three hundred and ten patients fulfilled the inclusion criteria, 526 (40%) of them showed LS >= 9 kPa and 316 (24%) cirrhosis (LS >= 14 kPa). The factors independently associated with significant fibrosis [adjusted odds ratio (95% confidence interval, P value) were the following: older age [1.04 (1.01-1.07), 0.002], daily alcohol intake > 50 g/day [1.58 (1.10-2.27), 0.013] and the length of HCV infection [1.03 (1.00 1.06), 0.023]]. A CD4 cell count lower than < 200 per mm(3) [1.67 (0.99-2.81), 0.053] and HCV genotype 4 [0.66 (0.42-1.02), 0.066] were marginally associated with LS >= 9 kPa. In conclusion, the prevalence of cirrhosis in HIV/HCV coinfected patients seems to be higher than previously reported in studies based on liver biopsy. Older age, alcohol consumption and lower CD4 cell counts are related with significant fibrosis. The latter association supports an earlier starting of antiretroviral therapy in this setting. PMID- 20002562 TI - Response to pegylated interferon and ribavirin in Asian American patients with chronic hepatitis C genotypes 1 vs 2/3 vs 6. AB - Chronic hepatitis C is generally underappreciated in Asian Americans, and most pivotal studies were conducted in western countries and only included a small numbers of Asian patients. Our goal was to examine and compare treatment outcomes in these patients with genotypes 1 vs 2/3 vs 6. We performed a retrospective cohort study of 167 consecutive treatment-naive Asian American patients treated with pegylated interferon (PEG IFN) plus ribavirin (RBV) at two community clinics in Northern California from 12/00 to 1/08. Primary outcome was sustained virological response rate by intention-to-treat analysis. The overall completion rate was 76%, and treatment adherence (completion of >= 75-80% PEG IFN + RBV dose for >= 75-80% of intended duration) was 74%. Significant depression was noted in only 4% of patients. Sustained virologic response in patients with genotype 6 treated for 48 weeks was similar to that seen in those with genotype 2/3 (74%vs 75%, P = 0.89) and significantly higher than those with genotype 1 (74%vs 49%, P = 0.016). On multivariate analysis inclusive of sex, age, body mass index (<= 25 vs > 25) and viral load, only treatment adherence and genotype (2/3 and 6 treated for 48 weeks) were found to be significant predictors of sustained virologic response. We conclude that significant depression is rare in Asian American patients (4%). Patients with genotype 6 treated for 48 weeks appear to have a similar treatment response rate as patients with genotype 2/3 and a significantly higher response rate than those with genotype 1. PMID- 20002563 TI - Relationship between functional polymorphism in the Aurora A gene and susceptibility of hepatocellular carcinoma. AB - Aurora A is considered a potential cancer susceptibility gene owing to overexpression or amplification of Aurora A gene that causes centrosome dysfunction, chromosome instability, tumourigenic transformation and checkpoint abnormalities. Functional coding region polymorphism F31I in the Aurora A gene has recently been shown to be associated with several human cancers, but its association with hepatocellular carcinoma (HCC) has yet to be investigated. Genetic polymorphism of Aurora A was investigated in 128 confirmed subjects with HCC and 128 cancer-free control subjects matched on age, gender, smoking and alcohol consumption by using a polymerase chain reaction-restriction fragment length polymorphism assay. Allele and genotype associations of Aurora A F31I polymorphism with HCC susceptibility were observed in comparisons between the patient and control samples (respectively; P = 0.005, P = 0.012). The proportion of the genotypes containing I31 allele in patients with HCC (39.8%) was significantly higher than that in patients without HCC (22.7%) (P = 0.003). The distribution F31I genotype was significantly associated with increased risk of HCC (P = 0.003, odds ratio = 2.26, 95% confidence interval = 1.31-3.90 for FI + II genotypes vs FF genotype). Our results suggest for the first time that the Aurora A F31I polymorphism may be a genetic susceptibility factor for HCC. PMID- 20002564 TI - Chemerin, vaspin and insulin resistance in chronic hepatitis C. AB - Adipocytokine profile seems to play a distinct role in the pathogenesis of chronic hepatitis C (CHC). Chemerin and vaspin are recently described adipocytokines with various suggested functions and potential to modulate inflammatory response and insulin resistance (IR). We assessed chemerin, vaspin and leptin serum concentration and studied their association with IR laboratory and morphological features in patients with hepatitis C. The study included 40 patients with hepatitis C and 20 healthy volunteers, similar in age and body mass index (43.6 +/- 11.6 vs 40.9 +/- 11.8 years and 25.0 +/- 4.1 vs 23.9 +/- 3.3 kg/m(2), respectively). Patients had to have a normal lipid profile, and diabetes was an exclusion criteria. Serum chemerin and leptin levels and IR were significantly higher in patients with hepatitis C when compared to the controls (P = 0.02, P = 0.02 and P = 0.02, respectively), whereas vaspin level was significantly decreased (P = 0.01). Serum chemerin was negatively associated with necro-inflammatory grade (r = (-0.49), P = 0.01). The lowest levels of serum chemerin were found in patients with moderate/severe inflammation (P = 0.03). Serum leptin tended to be up-regulated in patients with minimal inflammatory activity. Serum vaspin was higher, although not significantly, when fibrosis was more advanced. IR was positively associated with fibrosis stage (r = 0.33, P = 0.03). Serum chemerin and leptin were related to each other (r = 0.45, P = 0.02).Our findings support a complex interaction between the analysed adipokines and pathogenesis of inflammatory process in CHC. The role of chemerin and vaspin in pathogenesis of inflammatory response should be further investigated. PMID- 20002566 TI - Hepatitis E in HIV-positive patients in a low-endemic country. PMID- 20002567 TI - Hepatitis B virus: origin and evolution. AB - The pathogenesis of hepatitis B virus (HBV) is complex and it appears that molecular variants play a role in this process. HBV undergoes numerous rounds of error prone production within an infected host. The resulting quasispecies are heterogeneous and in the absence of archaeological records of past infection, the evolution of HBV can only be inferred indirectly from its epidemiology and by genetic analysis. This review gathered the controversies about the HBV origin and factors influencing its quasispecies. Also, it provided some evidence on how HBV genotypes correlated with human history and patterns of migration. It is our belief that this topic deserves further attention and thus it is likely that more critical research work will be performed to elucidate the unknown mechanisms and processes in this area. PMID- 20002565 TI - Role of treatment for depressive symptoms in relieving the impact of fatigue in HIV-HCV co-infected patients: ANRS Co13 Hepavih, France, 2006-2008. AB - Fatigue is a major component of quality of life (QOL) and is associated with depression in HIV-HCV co-infected individuals. We investigated whether treating depressive symptoms (DS) could mitigate the impact of fatigue on daily functioning in co-infected patients, even those at an advanced stage of disease. The analysis was conducted on enrollment data of 328 HIV-HCV co-infected patients recruited in the French nationwide ANRS CO 13 HEPAVIH cohort. Data collection was based on medical records and self-administered questionnaires which included items on socio-behavioural data, the fatigue impact scale (FIS) in three domains (cognitive, physical and social functioning), depressive symptoms (CES-D classification) and use of treatments for depressive symptoms (TDS). After multiple adjustment for gender and unemployment, CD4 cell count <200 per mm(3) was associated with a negative impact of fatigue on the physical functioning dimension (P = 0.002). A higher number of symptoms causing discomfort significantly predicted a higher impact of fatigue on all three dimensions (P < 0.001). This was also true for patients with DS receiving TDS when compared with those with no DS but receiving TDS. A significant decreasing linear trend (P < 0.001) of the impact of fatigue was found across the categories 'DS/TDS', 'DS/no TDS', 'no DS/TDS' and 'no DS/no TDS'. Despite limitations related to the cross sectional nature of this study, our results suggest that routine screening and treatment for DS can reduce the impact of fatigue on the daily functioning of HIV HCV co-infected patients and relieve the burden of their dual infection. PMID- 20002568 TI - Verification of an Edwardsiella ictaluri-specific diagnostic PCR. AB - AIMS: To verify the specificity of a PCR assay for the identification and diagnosis of Edwardsiella ictaluri. METHODS AND RESULTS: An Edwardsiella ictaluri specific PCR assay was developed utilizing two features of the ribosomal DNA gene clusters. The first feature is the presence of two ribosomal gene clusters located in tandem to one another (the inter-ribosomal spacer, IRS). This characteristic is present in the Edwardsiella genus but absent in the other sequenced members of the Enterobacteriaceae. The second feature is the presence of an intervening sequence (IVS) in the 23S rRNA gene of Edw. ictaluri. To verify the specificity of this assay, we tested genomic DNA from a variety of bacterial species. The IVS/IRS PCR assay results in an c. 2000-bp product from all Edw. ictaluri isolates tested, but not from any other species including Edwardsiella tarda. CONCLUSIONS: The IVS/IRS PCR assay is highly specific for Edw. ictaluri and useful as a tool for identifying this pathogen. SIGNIFICANCE AND IMPACT OF THE STUDY: This research verifies the specificity of PCR-based assay for Edw. Ictaluri, and we describe this assay as a highly versatile diagnostic tool for its identification. PMID- 20002569 TI - Isolation, evaluation and use of two strong, carbon source-inducible promoters from Corynebacterium glutamicum. AB - AIMS: To obtain strong, carbon source-inducible promoters useful for industrial applications of Corynebacterium glutamicum. METHODS AND RESULTS: DNA microarray and qRT-PCR enabled identification of the promoters of cgR_2367 (malE1) and cgR_2459 (git1) as strong, maltose- and gluconate-inducible promoters, respectively, in C. glutamicum. Promoter probe assays revealed that in the presence of the inducing sugars, PmalE1 and Pgit1, respectively, facilitated 3.4- and 4.2-fold increased beta-galactosidase activities compared to the same activity induced by glucose. In addition, PmalE1 was not functional in Escherichia coli, in which Pgit1 function was repressible, which enabled the cloning of a hitherto 'difficult-to-clone' heterologous gene of a lignocellulolytic enzyme, whose secretion was consequently induced by the carbon sources. CONCLUSIONS: PmalE1 and Pgit1 are strong, carbon source-inducible promoters of C. glutamicum whose characteristics in E. coli are integral to the secretion ability of C. glutamicum to secrete lignocellulolytic enzyme. SIGNIFICANCE AND IMPACT OF THE STUDY: Corynebacterium glutamicum, like its counterpart industrial workhorses E. coli and Bacillus subtilis, does exhibit strong, carbon source-inducible promoters, and the functionality of two of which was demonstrated in this study. While this study may be most relevant in the ongoing efforts to establish technologies of the biorefinery, it should also be of interest to general microbiologists exploring the versatility of industrial micro-organisms. In so doing, the study should impact future advances in industrial microbiology. PMID- 20002570 TI - Cloning, expression and characterization of attachment-invasion locus protein (Ail) of Yersinia enterocolitica and its utilization in rapid detection by immunoassays. AB - AIMS: Rapid detection of pathogenic Yersinia enterocolitica isolates by using antisera raised against recombinant attachment-invasion locus (Ail) protein. METHODS AND RESULTS: The complete gene (471 bp) encoding for the Ail protein was amplified by PCR and cloned in pQE 30 UA vector. The recombinant clones were selected by polymerase chain reaction (PCR). Recombinant protein was expressed using induction with 1 mmol l(-1) final concentration of isopropylthiogalactoside (IPTG). Polyclonal antibodies were raised in mice against this purified recombinant protein. An indirect plate ELISA was standardized based on rAil protein for the detection of Y. enterocolitica. Western blot analysis with the sera raised against recombinant Ail protein exhibited reaction at 17 kDa region of the native Ail protein present in pathogenic Y. enterocolitica standard strains and strains isolated from pork samples suggesting that the antigenicity of recombinant Ail protein was similar to that of native Ail protein. Nonpathogenic Y. enterocolitica and the other species of Yersinia, namely, Y. pseudotuberculosis, Y. intermedia, Y. kristenseni, Y. fredrickseni and also the Enterobacteriaceae organisms tested were not found reacting to polyclonal antisera against this recombinant Ail protein. CONCLUSION: The antibodies raised against recombinant Ail protein could specifically identify pathogenic Y. enterocolitica strains both by indirect plate ELISA and Western blot immunoassay. SIGNIFICANCE AND IMPACT OF THE STUDY: The method developed in this study may find application in the detection of pathogenic Y. enterocolitica not only from food and environmental samples but also from clinical samples. PMID- 20002571 TI - Method for bacteriophage isolation against target Campylobacter strains. AB - AIMS: Poultry meat is considered a major source of Campylobacter. This micro aerobic bacterium is commonly responsible for foodborne illness. This work focuses on the isolation of Campylobacter coli lytic bacteriophages (phages) against target C. coli strains. METHODS AND RESULTS: A method involving the enrichment of free-range chicken samples in a broth containing the target C. coli strains and salts (CaCl(2) and MgSO(4)) was used for phage isolation. This method allowed the isolation of 43 phages that were active against 83% of the C. coli strains used in the isolation procedure. Approximately 65% of the phages were also effective against Campylobacter jejuni strains. CONCLUSIONS: The use of target pathogens in the phage isolation step improves the likelihood of detecting and isolating phages for the control of these specific strains. SIGNIFICANCE AND IMPACT OF THE STUDY: This technique will be valuable in the context of phage therapy for enriching for phages that are active against specifically identified strains of bacteria, for example from a food poisoning outbreak or epidemic strains resistant to multiple antibiotics. In these situations, using the conventional methods for searching for bacteriophages active for these particular strains can be a time-consuming, if not an unsuccessful process. Using the isolation method described in this manuscript, the particular strains can be added to the enrichment broth increasing the probability of finding phages against them. Therefore, it will shorten the time needed for seeking phages able to lyse target strains, which in most of the cases, because of the rapid increase in antimicrobial-resistant bacteria, is of crucial importance. PMID- 20002572 TI - Production of single cell protein using waste capsicum powder produced during capsanthin extraction. AB - AIMS: To produce single cell protein (SCP) by using waste capsicum powder produced during capsanthin extraction as a substrate. METHODS AND RESULTS: The extraction [CPM (capsicum powder medium)] from waste capsicum powder was used as culture medium to cultivate four yeast strains. The main composition of CPM was determined. The average concentration of total sugar, total nitrogen and phosphorous of CPM were 16.3, 3.7 g l(-1) and 785.4 mg l(-1), respectively. Four yeast strains were cultured in two CPMs, and the cell mass, protein content of cells and specific growth rate of cells were determined. Addition of corn steep liquor significantly increased the cell mass production. Presence of capsaicin in CPM did not show inhibition of cell growth of yeast tested. CONCLUSIONS: CPM contained sufficient nutrients and could be used as a good medium to produce SCP. Candida utilis 1769 was chosen as the biomass producer because of its highest SCP formation (6.8 g l(-1)) and higher specific growth rate (0.12 h(-1)). The amino acid composition of its protein was well balanced. SIGNIFICANCE AND IMPACT OF THE STUDY: Utilization of waste capsicum powder can reduce environmental pollution and increase protein supply for animal feed. PMID- 20002573 TI - The purB gene controls rhizosphere colonization by Pantoea agglomerans. AB - AIMS: To isolate the rhizosphere competence-defective transposon Tn5 mutant of Pantoea agglomerans NBRISRM (SRM) and to identify the gene causing defect in its root colonization ability. METHODS AND RESULTS: From over 5000 clones containing Tn5, one mutant P. agglomerans NBRISRMT (SRMT) showing 6 log units less colonization when compared with SRM, after 30 days in sand-nonsterilized soil assay system was selected for further work to determine the effects of the mutation on rhizosphere competence. Southern hybridization analysis of restricted genomic DNA of SRMT demonstrated that the mutant had a single Tn5 insert. SRM increased in titre to about 2 x 10(8) CFU g(-1) root, compared with the indigenous bacterial population of heterotrophs of about 5 x 10(7) CFU g(-1) root. In contrast, 30 days later, the titre value of SRMT was almost undetectable at 1 x 10(2) CFU g(-1) root, demonstrating its inability to survive and colonize the rhizosphere. Sequencing of the flanking region of the Tn5 mutant revealed that Tn5 disrupted the purB gene. CONCLUSIONS: A defect in the colonization phenotype of the SRMT was attributed to the disruption in adenylosuccinate lyase (EC 4.3.2.2) which is encoded by the pur B gene and is required for rhizosphere colonization in P. agglomerans. Significantly less exopolysaccharide and biofilm was formed by SRMT when compared to SRM, because of the disruption of the purB gene. SIGNIFICANCE AND IMPACT OF THE STUDY: This work provides the first evidence for a functional role of purB gene in rhizosphere competence and root colonization by any rhizobacteria. PMID- 20002574 TI - Optimization of exopolysaccharide production from Armillaria mellea in submerged cultures. AB - AIMS: To study the optimization of submerged culture conditions for exopolysaccharide (EPS) production by Armillaria mellea in shake-flask cultures and also to evaluate the performance of an optimized culture medium in a 5-l stirred tank fermenter. METHODS AND RESULTS: Shake flask cultures for EPS optimal nutritional production contained having the following composition (in g l(-1)): glucose 40, yeast extract 3, KH(2)PO(4) 4 and MgSO(4) 2 at an optimal temperature of 22 degrees C and an initial of pH 4.0. The optimal culture medium was then cultivated in a 5-l stirred tank fermenter at 1 vvm (volume of aeration per volume of bioreactor per min) aeration rate, 150 rev min(-1) agitation speed, controlled pH 4.0 and 22 degrees C. In the optimal culture medium, the maximum EPS production in a 5-l stirred tank fermenter was 588 mg l(-1), c. twice as great as that in the basal medium. The maximum productivity for EPS (Q(p)) and product yield (Y(P/S)) were 42.02 mg l(-1) d(-1) and 26.89 mg g(-1), respectively. CONCLUSIONS: The optimal culture conditions we proposed in this study enhanced the EPS production of A. mellea from submerged cultures. SIGNIFICANCE AND IMPACT OF THE STUDY: The optimal culturing conditions we have found will be a suitable starting point for a scale-up of the fermentation process, helping to develop the production of related medicines and health foods from A. mellea. PMID- 20002575 TI - Miniprimer PCR assay targeting multiple genes: a new rapid and reliable tool for genotyping Pantoea stewartii subsp. stewartii. AB - AIM: Development of a 'miniprimer' PCR assay for genotyping Pantoea stewartii subsp. stewartii, the causal agent of the Stewart's bacterial wilt on maize. METHODS AND RESULTS: Four 10-nucleotide (10-nt) 'miniprimer' sets were designed and evaluated in the presence of Titanium Taq DNA polymerase. Under optimal reaction conditions, the miniprimer pair Uni-BacF-10/Uni-BacR-10 reproducibly generated identical banding patterns among 10 strains of P. stewartii subsp. stewartii, different patterns from strains of P. stewartii subsp. indologenes, other Panteoa species, Clavibacter michiganensis, Pectobacterium spp., Pseudomonas spp. and other bacterial species. The amplicons of Pantoea stewartii subsp. stewartii were cloned and sequenced to identify genes or DNA fragments that are targeted by the miniprimer PCR assay. Of the 14 'clone types' identified, sequences of a 1.23-kb fragment had a 99.8% similarity to part of the Pantoea stewartii zeaxanthin diglucoside biosynthetic operon (AY166713). Other dominant cloned fragments included a 411-bp amplicon that exhibited 99.8% similarity to the psaU gene (syn:ysaU; GQ249669), a type III protein-secretion system complex of P. stewartii subsp. stewartii strain DC283, and a 548-bp fragment showed 63% homology to the Asp/Glu racemase encoding gene in Erwinia tasmaniensis strain ET1/99. CONCLUSION: The miniprimer PCR assay reported here is highly discriminatory and reproducible in genotyping Pantoea stewartii subsp. stewartii. SIGNIFICANCE AND IMPACT OF THE STUDY: This miniprimer PCR assay could be a new reliable and rapid tool for fingerprinting the Stewart's wilt pathogen of maize. PMID- 20002576 TI - Human cathelicidin peptide LL37 inhibits both attachment capability and biofilm formation of Staphylococcus epidermidis. AB - AIMS: The aim of this work was to investigate the possible effect of human cathelicidin antimicrobial peptide LL37 on biofilm formation of Staphylococcus epidermidis, a major causative agent of indwelling device-related infections. METHODS AND RESULTS: We performed initial attachment assay and biofilm formation solid surface assay in microtitre plates, as well as growth experiment in liquid medium using laboratory strain Staph. epidermidis ATCC35984. We found that already a low concentration of the peptide LL37 (1 mg l(-1)) significantly decreased both the attachment of bacteria to the surface and also the biofilm mass. No growth inhibition was observed even at 16 mg l(-1) concentration of LL37, indicating a direct effect of the peptide on biofilm production. CONCLUSIONS: As biofilm protects bacteria during infections in humans and allows their survival in a hostile environment, inhibition of biofilm formation by LL37 may have a key role to prevent bacterial colonization on indwelling devices. SIGNIFICANCE AND IMPACT OF THE STUDY: Our findings suggest that this host defence factor can be a potential candidate in prevention and treatment strategies of Staph. epidermidis infections in humans. PMID- 20002577 TI - A recombinant metalloprotease antigen of Vibrio vulnificus elicits protective antibodies in a murine model. AB - AIMS: To investigate whether Vibrio vulnificus metalloprotease (VvpE) can induce the production of specific anti-VvpE antibody to confer effective protection against Vibrio vulnificus infection and to evaluate the possibility of VvpE as a potential vaccine candidate against disease caused by V. vulnificus. METHODS AND RESULTS: The gene encoding the 65-kDa VvpE of V. vulnificus was amplified by PCR and cloned into the expression vector pET21(b). The recombinant VvpE of V. vulnificus was expressed in Escherichia coli BL21(DE3). This His(6)-tagged VvpE was purified and injected intramuscularly into mice to evaluate its ability to stimulate immune response. Specific antibody levels were measured by ELISA. The 75% protective efficacy of recombinant VvpE was evaluated by active immunization and intraperitoneal challenge with V. vulnificus in mice. CONCLUSIONS: The recombinant His(6)-tagged VvpE of V. vulnificus is capable of inducing high antibody response in mice to confer effective protection against lethal challenge with V. vulnificus. VvpE might be a potential vaccine candidate to against V. vulnificus infection. SIGNIFICANCE AND IMPACT OF THE STUDY: This study uses His(6)-tagged VvpE to act as vaccine that successfully induces effective and specific anti-VvpE antibody and offers an option for the potential vaccine candidate against V. vulnificus infection. PMID- 20002578 TI - Repeated unidirectional introgression towards Populus balsamifera in contact zones of exotic and native poplars. AB - As the evolutionary significance of hybridization is largely dictated by its extent beyond the first generation, we broadly surveyed patterns of introgression across a sympatric zone of two native poplars (Populus balsamifera, Populus deltoides) in Quebec, Canada within which European exotic Populus nigra and its hybrids have been extensively planted since the 1800s. Single nucleotide polymorphisms (SNPs) that appeared fixed within each species were characterized by DNA-sequencing pools of pure individuals. Thirty-five of these diagnostic SNPs were employed in a high-throughput assay that genotyped 635 trees of different age classes, sampled from 15 sites with various degrees of anthropogenic disturbance. The degree of admixture within sampled trees was then assessed through Bayesian clustering of genotypes. Hybrids were present in seven of the populations, with 2.4% of all sampled trees showing spontaneous admixture. Sites with hybrids were significantly more disturbed than pure stands, while hybrids comprised both immature juveniles and trees of reproductive age. All three possible F1s were detected. Advanced-generation hybrids were consistently biased towards P. balsamifera regardless of whether hybridization had occurred with P. deltoides or P. nigra. Gene exchange between P. deltoides and P. nigra was not detected beyond the F1 generation; however, detection of a trihybrid demonstrates that even this apparent reproductive isolation does not necessarily result in an evolutionary dead end. Collectively, results demonstrate the natural fertility of hybrid poplars and suggest that introduced genes could potentially affect the genetic integrity of native trees, similar to that arising from introgression between natives. PMID- 20002579 TI - Ant genomics: strength and diversity in numbers. AB - A recent workshop held at the Arizona State University Center for Social Dynamics and Complexity gathered over 50 prominent researchers from around the globe to discuss the development of genomic resources for several ant species. Ants play crucial roles in many ecological niches and the sequencing of several ant genomes promises to elucidate topics ranging from the genetic basis for social complexity, longevity and behaviour to systems biology and the identification of novel antimicrobial compounds. Unlike other species, most ant genomes are being generated by individual labs and small collaborations without the annotation and computational resources that support prominent model organism genome databases such those for the fruitfly and roundworm. Attendees summarized their current progress and future plans for several ant genomes and discussed how best to coordinate the analysis and annotation of ant sequences to benefit the broad research interests of the social insect community. PMID- 20002580 TI - Association between Wolbachia and Spiroplasma within Drosophila neotestacea: an emerging symbiotic mutualism? AB - Interspecific mutualism can evolve when specific lineages of different species tend to be associated with each other from one generation to the next. Different maternally transmitted endosymbionts occurring within the same cytoplasmic lineage fulfil this requirement. Drosophila neotestacea is infected with maternally transmitted Wolbachia and Spiroplasma, which are cotransmitted at high frequency in natural populations. Molecular phylogenetic evidence indicates that both endosymbionts have been present in D. neotestacea for considerable evolutionary periods. Thus, conditions are suitable for the evolution of mutualism between them. In support of this possibility, there is a significant positive association between Wolbachia and Spiroplasma infection in many samples of D. neotestacea from natural populations. Theoretically, such a positive association can result from either mutualism between these endosymbionts or recent spread. Collections from present-day populations suggest that recent spread and mutualism have both operated to generate the positive association between Wolbachia and Spiroplasma. If selection acts on the combination of these two endosymbionts, they may be in the early stages of evolution of a more complex, cooperative association. PMID- 20002581 TI - Clonal expansion of the Belgian Phytophthora ramorum populations based on new microsatellite markers. AB - Co-existence of both mating types A1 and A2 within the EU1 lineage of Phytophthora ramorum has only been observed in Belgium, which begs the question whether sexual reproduction is occurring. A collection of 411 Belgian P. ramorum isolates was established during a 7-year survey. Our main objectives were genetic characterization of this population to test for sexual reproduction, determination of population structure, evolution and spread, and evaluation of the effectiveness and impact of control measures. Novel, polymorphic simple sequence repeat (SSR) markers were developed after screening 149 candidate loci. Eighty isolates of P. ramorum, broadly representing the Belgian population, were analyzed using four previously described and three newly identified polymorphic microsatellite loci as well as amplified fragment length polymorphisms. SSR analysis was most informative and was used to screen the entire Belgian population. Thirty multilocus genotypes were identified, but 68% of the isolates belonged to the main genotype EU1MG1. Although accumulated mutation events were detected, the overall level of genetic diversity within the Belgian isolates of P. ramorum appears to be limited, indicating a relatively recent clonal expansion. Based on our SSR analysis there is no evidence of sexual recombination in the Belgian population of P. ramorum. Metalaxyl use decreased the genetic diversity of P. ramorum until 2005, when the majority of the isolates had become resistant. Most genotypes were site-specific and despite systematic removal of symptomatic and neighbouring plants, some genotypes were detected over a period of several years at a single site, sometimes discontinuously, indicating (latent) survival of the pathogen at those sites. PMID- 20002582 TI - Establishment and expansion of Lake Malawi rock fish populations after a dramatic Late Pleistocene lake level rise. AB - Major environmental events that fragment populations among multiple island habitats have potential to drive large-scale episodes of speciation and adaptive radiation. A recent palaeolimnological study of sediment cores indicated that Lake Malawi underwent major climate-driven desiccation events 75,000-135,000 years ago that lowered the water level to at least 580 m below the present state and severely reduced surface area. After this period, lake levels rose and stabilized, creating multiple discontinuous littoral rocky habitats. Here, we present evidence supporting the hypothesis that establishment and expansion of isolated philopatric rock cichlid populations occurred after this rise and stabilization of lake level. We studied the Pseudotropheus (Maylandia) species complex, a group with both allopatric and sympatric populations that differ in male nuptial colour traits and tend to mate assortatively. Using coalescent analyses based on mitochondrial DNA, we found evidence that populations throughout the lake started to expand and accumulate genetic diversity after the lake level rise. Moreover, most haplotypes were geographically restricted, and the greatest genetic similarities were typically among sympatric or neighbouring populations. This is indicative of limited dispersal and establishment of assortative mating among populations following the lake level rise. Together, this evidence is compatible with a single large-scale environmental event being central to evolution of spatial patterns of genetic and species diversity in P. (Maylandia) and perhaps other Lake Malawi rock cichlids. Equivalent climate driven pulses of habitat formation and fragmentation may similarly have contributed to observed rapid and punctuated cladogenesis in other adaptive radiations. PMID- 20002583 TI - Cryptic differences in dispersal lead to differential sensitivity to habitat fragmentation in two bumblebee species. AB - Habitat loss has led to fragmentation of populations of many invertebrates, but social hymenopterans may be particularly sensitive to habitat fragmentation due to their low effective population sizes. The impacts of fragmentation depend strongly on dispersal abilities, but these are difficult to quantify. Here, we quantify and compare dispersal abilities of two bumblebee species, Bombus muscorum and Bombus jonellus, in a model island system. We use microsatellites to investigate population genetic structuring, dispersal and spatial patterns in genetic diversity. Populations of both species showed significant structuring, and isolation by distance, but this was markedly greater in B. muscorum (theta = 0.13) than in B. jonellus (theta = 0.034). This difference could reflect a higher effective population size in B. jonellus compared to B. muscorum, but this is not consistent with the observed abundance of the two species. We argue that it is more likely that B. jonellus has a higher propensity to disperse than B. muscorum. This will influence their relative susceptibility to habitat fragmentation and may in part explain differential declines of mainland populations of these and other bumblebee species. PMID- 20002584 TI - Out in the cold: genetic variation of Nothofagus pumilio (Nothofagaceae) provides evidence for latitudinally distinct evolutionary histories in austral South America. AB - Nothofagus pumilio is the dominant and almost ubiquitous tree species in mountainous environments of temperate South America. We used two types of molecular markers (cpDNA and isozymes) to evaluate the effects of the Paleogene paleogeography of Patagonia and more recent climatic oscillations of the Neogene on such cold-tolerant species' genetic makeup. Phylogeographic analysis on sequences of three cpDNA non-coding regions at 85 populations yielded two latitudinally disjunct monophyletic clades north and south of c. 42 degrees S containing 11 and three haplotypes, respectively. This indicates a long-lasting vicariant event due to the presence of an extended open paleobasin at mid latitudes of Patagonia. Also distribution patterns of cpDNA haplotypes suggest regional spread following stepping-stone models using pre-Cenozoic mountains as corridors. Comparable genetic diversity measured along 41 sampled populations using seven polymorphic isozyme loci provides evidence of local persistence and spread from multiple ice-free locations. In addition, significantly higher heterozygosity and allelic richness at high latitudes, i.e. in areas of larger glacial extent, suggest survival in large and isolated refugia. While, higher cpDNA diversity in lower latitudes reflects the complex orogeny that historically isolated northern populations, lower isozyme diversity and reduced F(ST) values provide evidence of local glacial survival in numerous small locales. Therefore, current genetic structure of N. pumilio is the result of regional processes which took place during the Tertiary that were enhanced by contemporary local effects of drift and isolation in response to Quaternary climatic cycles. PMID- 20002585 TI - Mercury-associated DNA hypomethylation in polar bear brains via the LUminometric Methylation Assay: a sensitive method to study epigenetics in wildlife. AB - In this paper we describe a novel approach that may shed light on the genomic DNA methylation of organisms with non-resolved genomes. The LUminometric Methylation Assay (LUMA) is permissive for genomic DNA methylation studies of any genome as it relies on the use of methyl-sensitive and -insensitive restriction enzymes followed by polymerase extension via Pyrosequencing technology. Here, LUMA was used to characterize genomic DNA methylation in the lower brain stem region from 47 polar bears subsistence hunted in central East Greenland between 1999 and 2001. In these samples, average genomic DNA methylation was 57.9% +/- 6.69 (SD; range was 42.0 to 72.4%). When genomic DNA methylation was related to brain mercury (Hg) exposure levels, an inverse association was seen between these two variables for the entire study population (P for trend = 0.17). After dichotomizing animals by gender and controlling for age, a negative trend was seen amongst male animals (P for trend = 0.07) but no associations were found in female bears. Such sexually dimorphic responses have been found in other toxicological studies. Our results show that genomic DNA methylation can be quantitatively studied in a highly reproducible manner in tissue samples from a wild organism with a non-resolved genome. As such, LUMA holds great promise as a novel method to explore consequential questions across the ecological sciences that may require an epigenetic understanding. PMID- 20002586 TI - Comparative phylogeography of five avian species: implications for Pleistocene evolutionary history in the Qinghai-Tibetan plateau. AB - Pleistocene climate fluctuations have shaped the patterns of genetic diversity observed in extant species. In contrast to Europe and North America where the effects of recent glacial cycles on genetic diversity have been well studied, the genetic legacy of the Pleistocene for the Qinghai-Tibetan (Tibetan) plateau, a region where glaciation was not synchronous with the North Hemisphere ice sheet maxima, remains poorly understood. Here, we compared the phylogeographical patterns of five avian species on the Qinghai-Tibetan plateau by three mitochondrial DNA fragments: the Tibetan snow finch (Montifringilla adamsi), the Blanford's snow finch (Pyrgilauda blanfordi), the horned lark (Eremophila alpestris), the twite (Carduelis flavirostris) and the black redstart (Phoenicurus ochruros). Our results revealed the three species mostly distributed on the platform region of the plateau that experienced population expansion following the retreat of the extensive glaciation period (0.5-0.175 Ma). These results are at odds with the results from avian species of Europe and North America, where population expansions occurred after Last Glacial Maximum (LGM, 0.023-0.018 Ma). A single refugium was identified in a restricted semi-continuous area around the eastern margin of the plateau, instead of multiple independent refugia for European and North American species. For the other two species distributed on the edges of the plateau (the twite and black redstart), populations were maintained at stable levels. Edge areas are located on the eastern margin, which might have had little or no ice cover during the glaciation period. Thus, milder climate may have mitigated demographic stresses for edge species relative to the extremes experienced by platform counterparts, the present-day ranges of which were heavily ice covered during the glaciation period. Finally, various behavioural and ecological characteristics, including dispersal capacities, habitat preference and altitude specificity along with evolutionary history might have helped to shape different phylogeographical structures appearing in these five species. PMID- 20002587 TI - Heterothallism in Saccharomyces cerevisiae isolates from nature: effect of HO locus on the mode of reproduction. AB - Understanding the evolution of sex and recombination, key factors in the evolution of life, is a major challenge in biology. Studies of reproduction strategies of natural populations are important to complement the theoretical and experimental models. Fungi with both sexual and asexual life cycles are an interesting system for understanding the evolution of sex. In a study of natural populations of yeast Saccharomyces cerevisiae, we found that the isolates are heterothallic, meaning their mating type is stable, while the general belief is that natural S. cerevisiae strains are homothallic (can undergo mating-type switching). Mating-type switching is a gene-conversion process initiated by a site-specific endonuclease HO; this process can be followed by mother-daughter mating. Heterothallic yeast can mate with unrelated haploids (amphimixis), or undergo mating between spores from the same tetrad (intratetrad mating, or automixis), but cannot undergo mother-daughter mating as homothallic yeasts can. Sequence analysis of HO gene in a panel of natural S. cerevisiae isolates revealed multiple mutations. Good correspondence was found in the comparison of population structure characterized using 19 microsatellite markers spread over eight chromosomes and the HO sequence. Experiments that tested whether the mating type switching pathway upstream and downstream of HO is functional, together with the detected HO mutations, strongly suggest that loss of function of HO is the cause of heterothallism. Furthermore, our results support the hypothesis that clonal reproduction and intratetrad mating may predominate in natural yeast populations, while mother-daughter mating might not be as significant as was considered. PMID- 20002588 TI - Plastidial glycolysis in developing Arabidopsis embryos. AB - During oilseed embryo development, carbon from sucrose is utilized for fatty acid synthesis in the plastid. The role of plastidial glycolysis in Arabidopsis embryo oil accumulation was investigated. Genes encoding enolases (ENO) and phosphoglyceromutases (PGlyM) were identified, and activities and subcellular locations were established by expression of recombinant and green fluorescent protein (GFP)-fusion proteins. Mutant Arabidopsis plants lacking putative plastidial isoforms were characterized with respect to isoform composition and embryo oil content. In the developing embryo, ENO1 and ENO2 account for most or all of the plastidial and cytosolic ENO activity, respectively, and PGLYM1 accounts for most or all of the plastidial PGlyM activity. The eno1 and pglym1 mutants, in which plastidic ENO and PGlyM activities were undetectable, had wild type amounts of seed oil at maturity. It is concluded that although plastids of developing Arabidopsis embryos have the capacity to carry out the lower part of the glycolytic pathway, the cytosolic glycolytic pathway alone is sufficient to support the flux from 3-phosphoglycerate to phosphoenolpyruvate required for oil production. The results highlight the importance for oil production of translocators that facilitate interchange of glycolytic intermediates between the cytosol and the plastid stroma. PMID- 20002589 TI - Out-of-pocket prices of opioid analgesics in the United States, 1999-2004. AB - OBJECTIVE: To determine the out-of-pocket prices of common opioid analgesics by medication, drug coverage, region, and year. DESIGN: Retrospective cohort study using 1999-2004 data from the Medical Expenditure Panel Survey and the Medicare Current Beneficiary Survey. SETTING: U.S. civilian noninstitutionalized population. PATIENTS: Adults not enrolled in Medicaid who filled prescriptions for opioid analgesics between 1999 and 2004. OUTCOME MEASURES: Prices of prescribed analgesics were collected from receipts, medication containers, patient recall, and administrative records (N = 20,926 and 31,500, respectively). RESULTS: Average out-of-pocket price of an opioid analgesic prescription was around $10, but the estimate is potentially misleading: A typical adult patient without drug coverage paid $12.86-$61.60 to fill his or her analgesic prescription, depending on medication. The extended-release formulations cost more than double the immediate release prices. For the analgesics studied, drug coverage lowered out-of-pocket prices by 50-85%, while market prices increased at a rate of 5.7-9% per year with little regional variation. Data did not include prices for medications not prescribed or prescribed, but not acquired. CONCLUSIONS: Independent of the diagnosis, patients' out-of-pocket price for prescribed analgesics fluctuated freely in the United States across time, region, and coverage status. These fluctuations potentially distort the delivery of effective pain management and further burden an already afflicted population. PMID- 20002590 TI - Opioid analgesics for pain control: wisconsin physicians' knowledge, beliefs, attitudes, and prescribing practices. AB - OBJECTIVE: Opioid analgesics are the drugs of choice for the treatment of moderate to severe acute and cancer pain. Although their role in the management of chronic pain not related to cancer is controversial, there is increasing evidence for their benefit in certain patient populations. DESIGN: A 32-item survey to assess Wisconsin physicians' knowledge, beliefs, and attitudes toward opioid analgesic use was mailed to 600 randomly selected licensed physicians, resulting in a 36% response rate. RESULTS: Half of the respondents considered diversion a moderate or severe problem in Wisconsin. A majority considered addiction to be a combination of physiological and behavioral characteristics, rather than defining it solely as a behavioral syndrome. Most physicians felt it lawful and acceptable medical practice to prescribe opioids for chronic cancer pain, but only half held this view if the pain was not related to cancer. Fewer physicians considered such prescribing as lawful and generally accepted medical practice if the patient had a history of substance abuse. About two-thirds of physicians were not concerned about being investigated for their opioid prescribing practices, but some admitted that fear of investigation led them to lower the dose prescribed, limit the number of refills, or prescribe a Schedule III or IV rather than a Schedule II opioid. CONCLUSION: Wisconsin physicians who responded to this survey held many misconceptions about the prescribing of opioids. Such views, coupled with a lack of knowledge about laws and regulations governing the prescribing of controlled substances, may result in inadequate prescribing of opioids with resultant inadequate management of pain. PMID- 20002591 TI - Sex Differences in Pain and Pain-Related Disability among Primary Care Patients with Chronic Musculoskeletal Pain. AB - BACKGROUND: Although previous research suggests women report more severe pain than men, evidence for sex-related differences in pain-related disability is conflicting. Also, the impact of psychological factors on sex differences in disability is uncertain. OBJECTIVE: The purpose of this study is to assess sex differences in pain-related disability and evaluate whether they are accounted for by psychological factors. METHODS: Analysis of baseline data from the Stepped Care for Affective disorders and Musculoskeletal Pain study. Participants included 241 male and 249 female primary care patients with moderately severe persistent pain of the back, hip, or knee. Multivariable log-linear models were used to determine the association between sex and pain-related disability and whether sex differences persisted after adjustment for psychiatric comorbidity and potential psychological mediators. RESULTS: Compared with men, women reported worse pain intensity, greater pain-related interference with function, and more disability days due to pain. They also had worse depression, anxiety, and self efficacy. Sex differences in pain interference with function and pain disability days remained significant in multivariable models. Depression, poor self efficacy, and fear of reinjury were independently associated with disability in both men and women. CONCLUSIONS: Women report greater pain-related disability than do men, even after controlling for depression, anxiety, and other psychological factors. Pain management strategies that target functional disability may be particularly important in the treatment of women with pain. PMID- 20002592 TI - Predictors of pain outcomes in patients with chronic musculoskeletal pain co morbid with depression: results from a randomized controlled trial. AB - OBJECTIVE: The combination of chronic musculoskeletal pain and depression is associated with worse clinical outcomes than either condition alone. In this study, we report the predictors of pain intensity and activity interference in primary care patients with co-morbid pain and depression. METHODS: This is a secondary data analysis of the 250 persons who participated in a randomized clinical trial designed to test the effectiveness of 12 weeks of optimized antidepressant therapy for both depression and pain. Using multivariate linear regression analysis, we assessed the predictive value of baseline self-efficacy, fear of movement, pain beliefs, and demographic and clinical factors on 3-month Graded Chronic Pain Scale pain intensity and activity interference outcomes. RESULTS: In the full model, significant sociodemographic predictors of less activity interference included being non-white (beta-5.8, P = 0.04) and being employed (beta-13.3, P < 0.0001). The latter was also predictive of less pain intensity (beta-5.6, P = 0.01). As expected, the optimized antidepressant treatment arm was associated with improved outcomes (pain intensity: beta-3.7, P = 0.0005 and activity interference: beta-6.4, P = 0.01). Whereas stronger perceived pain control (beta 3.6, P = 0.01) was associated with greater activity interference, higher degree of fear of movement (or fear avoidance) predicted greater pain intensity (beta 0.46, P = 0.04) and activity interference (beta 0.57, P = 0.05). Neither the location (low back vs hip/knee) nor duration of pain were predictive of pain intensity or interference outcomes. CONCLUSION: The findings are consistent with a bio-psychosocial model, implicating the need to consider the impact of sociodemographic variables and pain-related beliefs and cognition on pain-related outcomes for patients with co-morbid musculoskeletal pain and depression. PMID- 20002593 TI - Importance of pelvic muscle tenderness evaluation in women with chronic pelvic pain. AB - OBJECTIVE: To determine the prevalence of pelvic muscle tenderness in women with chronic pelvic pain (CPP) and to assess the importance of evaluating muscle tenderness in such women. DESIGN: Observational study of 48 healthy female volunteers and 108 women with CPP, who were clinically evaluated for pelvic muscle tenderness by two researchers blinded to all clinical data. RESULTS: The frequency of clinically detected pelvic muscle tenderness was significantly higher in women with CPP than in healthy volunteers (58.3% vs 4.2%, P < 0.001). Among women with CPP, those with pelvic muscle tenderness had higher Beck Depression Index scores (22 [6-42] vs 13 [3-39], P = 0.02) and higher rates of dyspareunia (63.5% [40/63] vs 28.9% [13/45], P < 0.004) and constipation (46.0% [29/63] vs 26.7% [12/45], P = 0.05) than those without pelvic muscle tenderness. CONCLUSION: Tenderness of pelvic muscles was highly prevalent among women with CPP and was associated with higher BDI scores and higher rates of dyspareunia and constipation. Determination of pelvic muscle tenderness may help in identifying women who require more intense treatment for CPP. PMID- 20002594 TI - An open pilot study assessing the benefits of quetiapine for the prevention of migraine refractory to the combination of atenolol, nortriptyline, and flunarizine. AB - BACKGROUND: Migraine is a prevalent neurological disorder. Although prevention is the core of treatment for most, some patients are refractory to standard therapies. Accordingly, the aim of this study was to evaluate the use of Quetiapine (QTP) in the preventive treatment of refractory migraine, defined as previous unresponsiveness to the combination of atenolol, nortriptyline, and flunarizine. METHODS: Thirty-four consecutive patients (30 women and 4 men) with migraine (ICHD-II), fewer than 15 days of headache per month, and not overusing symptomatic medications were studied. All participants had failed to the combination of atenolol (60 mg/day), nortriptyline (25 mg/day), and flunarizine (3 mg/day). Failure was defined as <50% reduction in attack frequency after 10 weeks of treatment. After other medications were discontinued, QTP was initiated at a single daily dose of 25 mg, and then titrated to 75 mg. After 10 weeks, headache frequency, consumption of rescue medications, and adverse events were analyzed. RESULTS: Twenty-nine patients completed the study. Three patients withdrew and two were lost to follow-up. Among those who completed, 22 (75.9%; 64.7% of the intention-to-treat population) had greater than 50% headache reduction. The mean frequency of migraine days decreased from 10.2 to 6.2 per month. Use of rescue medications decreased from 2.3 to 1.2 days/week. Adverse events were reported by nine (31%) patients. CONCLUSIONS: Although limited by the open design, this study provides pilot data to support the use of QTP in the preventive treatment of refractory migraine. Controlled studies are necessary to confirm these observations. PMID- 20002595 TI - Assessment of celiac plexus block and neurolysis outcomes and technique in the management of refractory visceral cancer pain. AB - OBJECTIVE: To assess demographic and clinical factors associated with celiac plexus neurolysis outcomes. DESIGN: Retrospective clinical data analysis. SETTING: A tertiary care, academic medical center. PATIENTS: Forty-four patients with terminal visceral (mostly pancreatic) cancer who failed conservative measures. Interventions. Fifty celiac plexus alcohol neurolytic procedures done for pain control after a positive diagnostic block. OUTCOME MEASURES: A successful treatment was predefined as >50% pain relief sustained for > or =1 month. The following variables were analyzed for their association with treatment outcome: age, gender, duration of pain, origin of tumor, opioid dose, type of radiological guidance used, single- vs double-needle approach, type of block (e.g., antero- vs retrocrural), immediate vs delayed neurolysis, volume of local anesthetic employed for both diagnostic and neurolytic blocks, and use of sedation. RESULTS: Those variables correlated with a positive outcome included lower opioid dose and the absence of sedation. Strong trends for a positive association with outcome were found for the use of computed tomography (vs fluoroscopy), and using <20 mL of local anesthetic for the diagnostic block. CONCLUSIONS: Celiac plexus neurolysis may provide intermediate pain relief to a significant percentage of cancer sufferers. Both careful selection of candidates based on clinical variables, and technical factors aimed at enhancing the specificity of blocks may lead to improved outcomes. PMID- 20002596 TI - Durability of therapeutic response to milnacipran treatment for fibromyalgia. Results of a randomized, double-blind, monotherapy 6-month extension study. AB - OBJECTIVE: To evaluate the durability of improvement and long-term efficacy of milnacipran treatment in fibromyalgia, to assess efficacy in patients re randomized from placebo to milnacipran, and to collect additional information on the tolerability and efficacy of long-term treatment with milnacipran. DESIGN: A total of 449 patients who successfully completed a 6-month lead-in study enrolled in this 6-month extension study (87.7% of eligible subjects). Patients initially receiving milnacipran 200 mg/day during the lead-in study were maintained at 200 mg/day (n = 209); patients initially assigned to placebo or milnacipran 100 mg/day were re-randomized (1:4) to either 100 mg/day (n = 48) or 200 mg/day (n = 192) of milnacipran for an additional 6 months of treatment. Efficacy assessments included visual analog scale pain ratings, Fibromyalgia Impact Questionnaire (FIQ) total score, and Patient Global Impression of Change (PGIC). RESULTS: Patients continuing on milnacipran demonstrated a sustained reduction in pain over the full 12-month period. Additional beneficial effects were also maintained, as indicated by the PGIC and FIQ. Patients initially assigned to either placebo or milnacipran 100 mg/day in the lead-in study and subsequently re randomized to milnacipran 200 mg/day in the extension study experienced further improvements in their mean pain scores, FIQ total scores, and PGIC ratings at 1 year. Milnacipran treatment was generally well tolerated. The most commonly reported newly emergent adverse event was nausea. CONCLUSIONS: In addition to confirming that milnacipran safely and effectively improves the multiple symptoms of fibromyalgia, these data indicate that milnacipran provides 1-year durable efficacy in this patient population. PMID- 20002597 TI - Opioids for the management of severe chronic nonmalignant pain in children: a retrospective 1-year practice survey in a children's hospital. AB - OBJECTIVE: This study reviewed opioid prescription for chronic severe nonmalignant pain in a multidisciplinary pediatric pain clinic. We looked at benefits and side effects of therapy, and compared our process of opioid prescription with the practice guidelines defined in adult literature. DESIGN: Descriptive retrospective practice survey. SETTING: Multidisciplinary pain clinic in a tertiary pediatric hospital. PATIENTS: During a 12-month period, 104 patients were seen in the clinic, of which 49 received an opioid as part of their pain management; 11 received an opioid chronically, defined as more than 3 months in this study, and 5 of these were still on opioid at the end of the study period although data on one patient are lacking as she had been transferred to an adult clinic. METHODS: Information about patients was obtained from chart review. OUTCOME MEASURES: Benefits and side effects of treatment, as well as the process of prescribing opioids and follow-up. RESULTS: In the four patients studied, there seem to be better pain control and improved function in these patients while on opioid therapy, despite minor side effects. We identified some areas of improvement in our practice and made recommendations for the use of opioids in pediatric chronic severe nonmalignant pain. CONCLUSIONS: The evolution of our four patients seems to be in favor of treatment with an opioid for severe chronic nonmalignant pain in certain pediatric patients, in the context of prescribing in a multidisciplinary pain clinic with a multisystem approach to pain management although more data are needed to know if such therapy is safe and beneficial on a longer-term basis. PMID- 20002598 TI - Malignancy as a possible complication of complex regional pain syndrome: a case report. AB - A synovial sarcoma presented in the knee of a young woman 20 years after the onset of pain which was attributed to complex regional pain syndrome (CRPS). Was this a chance occurrence, or could there be any link between the two conditions? Did the pain itself and the persistent inflammatory and immunological response to pain contribute to the development of malignancy, or could the malignancy have been present subclinically for many years and have contributed to the ongoing pain syndrome? This case report looks into the diagnosis of synovial sarcoma and CRPS and the relationship between the neurogenic inflammation seen in CRPS and that seen in malignancies. The diagnosis of CRPS is a diagnosis of exclusion. Constant vigilance of patients with this unpleasant condition is necessary. PMID- 20002599 TI - A comprehensive protocol to diagnose and treat pain of muscular origin may successfully and reliably decrease or eliminate pain in a chronic pain population. AB - OBJECTIVE: A comprehensive protocol is presented to identify muscular causes of regional pain syndromes utilizing an electrical stimulus in lieu of palpation, and combining elements of Prolotherapy with trigger point injections. METHODS: One hundred seventy-six consecutive patients were evaluated for the presence of muscle pain by utilizing an electrical stimulus produced by the Muscle Pain Detection Device. The diagnosis of "Muscle Pain Amenable to Injection" (MPAI), rather than trigger points, was made if pain was produced for the duration of the stimulation. If MPAI was found, muscle tendon injections (MTI) were offered to patients along with post-MTI physical therapy, providing neuromuscular electrical stimulation followed by a validated exercise program [1]. A control group, evaluated 1 month prior to their actual consultation/evaluation when muscle pain was identified but not yet treated, was used for comparison. RESULTS: Forty-five patients who met criteria completed treatment. Patients' scores on the Brief Pain Inventory decreased an average of 62%; median 70% (P < 0.001) for pain severity and 68%; median 85% (P < 0.001) for pain interference one month following treatment. These changes were significantly greater (P < 0.001) than those observed in the untreated controls. CONCLUSION: A protocol incorporating an easily reproducible electrical stimulus to diagnose a muscle causing pain in a region of the body followed by an injection technique that involves the entirety of the muscle, and post injection restoration of muscle function, can successfully eliminate or significantly reduce regional pain present for years. PMID- 20002600 TI - Differences in growth and physiological traits of two poplars originating from different altitudes as affected by UV-B radiation and nutrient availability. AB - Cuttings of Populus kangdingensis and Populus cathayana originating from altitudes of 3500 and 1500 m in southwestern China, respectively, were grown for one growing season in the field under ambient or ambient plus supplemental ultraviolet-B (UV-B) radiation with two levels of nutrients. In both species, enhanced UV-B radiation significantly increased UV-B absorbing compounds and guaiacol peroxidase (GPX) activity, while no significant effects were observed in photosynthetic pigments and proline content. On the other hand, cuttings grown with high-nutrient availability had larger leaf area, higher total biomass and GPX activity as well as higher water use efficiency (WUE) (as measured by stable carbon isotope composition, delta(13)C) when compared with low-nutrient conditions, while UV-B absorbing compounds and ascorbic acid (AsA) content significantly decreased. Differences in responses to enhanced UV-B radiation and nutrient availability were observed between the two species. Nutrient-induced increases in chlorophyll a, chlorophyll b and total chlorophyll as well as in carotenoids were greater in P. kangdingensis than in P. cathayana. In P. cathayana, enhanced UV-B radiation significantly decreased leaf area and total biomass, while it significantly increased WUE and ascorbate peroxidase (APX). In contrast, such changes were not observed in P. kangdingensis. In addition, the effects of enhanced UV-B radiation on leaf area, total biomass and UV-B absorbing compounds were closely related to the nutrient status. Our results indicated that P. kangdingensis, which originates from the altitude of 3500 m and is apparently adapted to low-nutrient and high-UV-B habitats, exhibits better tolerance to enhanced UV-B radiation and greater growth under low-nutrient availability than does P. cathayana originating from the altitude of 1500 m. PMID- 20002601 TI - ROS in biotic interactions. AB - Production of reactive oxygen species (ROS) is a hallmark of successful recognition of infection and activation of plant defenses. ROS play multifaceted signaling functions mediating the establishment of multiple responses and can act as local toxins. Controversy surrounds the origin of these ROS. Several enzymatic mechanisms, among them a plasma membrane NADPH oxidase and cell wall peroxidases, can be responsible for the ROS detected in the apoplast. However, high levels of ROS from metabolic origins and/or from downregulation of ROS-scavenging systems can also accumulate in different compartments of the plant cell. This compartmentalization could contribute to the specific functions attributed to ROS. Additionally, ROS interact with other signals and phytohormones, which could explain the variety of different scenarios where ROS signaling plays an important part. Interestingly, pathogens have developed ways to alter ROS accumulation or signaling to modify plant defenses. Although ROS have been mainly associated with pathogen attack, ROS are also detected in other biotic interactions including beneficial symbiotic interactions with bacteria or mycorrhiza, suggesting that ROS production is a common feature of different biotic interactions. Here, we present a comprehensive review describing the newer views in ROS signaling and function during biotic stress. PMID- 20002602 TI - Burkholderia species are ancient symbionts of legumes. AB - Burkholderia has only recently been recognized as a potential nitrogen-fixing symbiont of legumes, but we find that the origins of symbiosis in Burkholderia are much deeper than previously suspected. We sampled 143 symbionts from 47 native species of Mimosa across 1800 km in central Brazil and found that 98% were Burkholderia. Gene sequences defined seven distinct and divergent species complexes within the genus Burkholderia. The symbiosis-related genes formed deep Burkholderia-specific clades, each specific to a species complex, implying that these genes diverged over a long period within Burkholderia without substantial horizontal gene transfer between species complexes. PMID- 20002603 TI - The east-west-north colonization history of the Mediterranean and Europe by the coastal plant Carex extensa (Cyperaceae). AB - Coastal plants are ideal models for studying the colonization routes of species because of the simple linear distributions of these species. Carex extensa occurs mainly in salt marshes along the Mediterranean and European coasts. Variation in cpDNA sequences, amplified fragment length polymorphisms (AFLPs) and simple sequence repeats (SSRs) of 24 populations were analysed to reconstruct its colonization history. Phylogenetic relationships indicate that C. extensa together with the South American Carex vixdentata and the southern African Carex ecklonii form a monophyletic group of halophilic species. Analyses of divergence times suggest that early lineage diversification may have occurred between the late Miocene and the late Pliocene (Messinian crisis). Phylogenetic and network analyses of cpDNA variation revealed the monophyly of the species and an ancestral haplotype contained in populations of the eastern Mediterranean. The AFLP and SSR analyses support a pattern of variation compatible with these two lineages. These analyses also show higher levels of genetic diversity and differentiation in the eastern population group, which underwent an east-to-west Mediterranean colonization. Quaternary climatic oscillations appear to have been responsible for the split between these two lineages. Secondary contacts may have taken place in areas near the Ligurian Sea in agreement with the gene flow detected in Corsican populations. The AFLP and SSR data accord with the 'tabula rasa' hypothesis in which a recent and rapid colonization of northern Europe took place from the western Mediterranean after the Last Glacial Maximum. The unbalanced west-east vs. west-north colonization may be as a result of 'high density blocking' effect. PMID- 20002604 TI - Local selection and population structure in a deep-sea fish, the roundnose grenadier (Coryphaenoides rupestris). AB - Local populations within a species can become isolated by stochastic or adaptive processes, though it is most commonly the former that we quantify. Using presumably neutral markers we can assess the time-dependent process of genetic drift, and thereby quantify patterns of differentiation in support of the effective management of diversity. However, adaptive differences can be overlooked in these studies, and these are the very characteristics that we hope to conserve by managing neutral diversity. In this study, we used 16 hypothetically neutral microsatellite markers to investigate the genetic structure of the roundnose grenadier in the North Atlantic. We found that one locus was a clear outlier under directional selection, with F(ST) values much greater than at the remaining loci. Differentiation between populations at this locus was related to depth, suggesting directional selection, presumably acting on a linked locus. Considering only the loci identified as neutral, there remained significant population structure over the region of the North Atlantic studied. In addition to a weak pattern of isolation by distance, we identified a putative barrier to gene flow between sample sites either side of the Charlie Gibbs Fracture Zone, which marks the location where the sub-polar front crosses the Mid-Atlantic Ridge. This may reflect a boundary across which larvae are differentially distributed in separate current systems to some extent, promoting differentiation by drift. Structure due to both drift and apparent selection should be considered in management policy. PMID- 20002605 TI - Risk-adjusted assessment of incidence and quantity of blood use in acute-care hospitals in Japan: an analysis using administrative data. AB - BACKGROUND AND OBJECTIVES: Continuous monitoring of blood use and feedback on transfusions are effective in decreasing inappropriate blood transfusions. However, traditional methods of monitoring have practical challenges, such as the limited availability of experts and funding. Administrative data including a patient classification system may be employed for risk-adjusted assessment of hospital-wide blood use. MATERIALS AND METHODS: We conducted an audit of blood use at two hospitals and determined proportions of appropriate blood use at each hospital. We then used administrative data of 587,045 cases provided by 73 hospitals to develop two mathematical models to calculate risk-adjusted use of blood products. The first model is a logistic regression model to predict the percentage of transfused patients. Patient demographics, surgery and diagnostic groups were utilized as predictors of transfusion. The second model is a case-mix adjusted model which predicts hospital-wide use of units of blood products from the distribution of diagnosis-related groups. For each model, the observed to expected (O/E) ratio of blood use in each hospital was calculated. We compared resultant ratios with proportions of appropriate blood use in two of the hospitals studied. RESULTS: Both models showed good prediction abilities. O/E ratios calculated using the two models were relevant to proportions of appropriate transfusions. CONCLUSIONS: Risk-adjusted assessments of blood product use based on administrative data allow hospital-wide evaluation of transfusion use. Comparing blood use between different hospitals contributes toward establishing appropriate transfusion practices. PMID- 20002606 TI - Leukotrienes affect secretory function of ovarian cells in vitro. AB - The aim of this study was to determine which cells are the source of production and target for leukotriene (LTs) action within the bovine ovary. Luteal (CL, days 14-16 of the oestrous cycle), steroidogenic cells (LSC) and endothelial cells (LEC) of the bovine corpus luteum (CL), and granulosa cells (GC) were isolated enzymatically, cultured in a monolayer and incubated with LTC(4), LTB(4), Azelastine (an antagonist of LTC(4)) or Dapsone (an antagonist of LTB(4)). Then cells were collected for determination of mRNA expression for LT receptors (LTRs) and 5-lipoxygenase (5-LO) by real time RT-PCR, and media were collected for determination of prostaglandin (PG)E(2), F(2alpha), progesterone (P4; LSC only), endothelin-1 (ET-1; LEC only) and 17-beta oestradiol (E2; GC only). The greatest mRNA expression for LTR-II and 5-LO were found in LEC, whereas LTR-I mRNA expression did not differ among cell types. The level of PGE(2) increased after LTs treatment in each type of ovarian cell, excluding LTC(4) treatment in LEC. The secretion of PGF(2alpha) was also increased by LTs, but decreased after LTB(4) treatment of LSC. In GC cultures, both LTs stimulated E2 secretion; in LEC cultures, LTB(4) stimulated whereas LTC(4) inhibited P4 secretion; in LEC cultures, LTC(4) stimulated but LTB(4) inhibited ET-1 secretion. The results show that LTs are produced locally and are involved in PGs production/secretion in all examined cells (LSC, LEC and GC) of bovine ovary. Leukotriene treatment modulate secretion of E2, by GC, P4 by LSC and ET-1 by LEC, which indicates that LTs are involved in regulation of ovarian secretory functions. PMID- 20002607 TI - Effect of beta-carotene supply during close-up dry period on the onset of first postpartum luteal activity in dairy cows. AB - The aim of this study was to examine the effect of beta-carotene supply during the close-up dry period on the onset of first postpartum luteal activity in dairy cows. Twelve cows were supplied with 2000 mg of beta-carotene (20 g Rovimix((r)) beta-Carotene containing 10% beta-carotene; DSM Nutrition Japan K.K., Tokyo, Japan) by oral administration daily from day 21 before expected calving date to parturition. Fourteen cows (control) did not receive beta-carotene supplementation. Blood samples were obtained on days 21, 14 and 7 before expected calving date and on days 1, 7, 14, 21 postpartum. When the plasma progesterone concentration exceeded 1 ng/ml by day 21 postpartum, luteal activity was assumed to have been initiated. The result showed that serum beta-carotene concentrations in the beta-carotene cows were higher than in the control cows during the experimental period (p < 0.01). The number of cows with the onset of luteal activity by day 21 postpartum was 9/12 in the beta-carotene cows and 4/14 in the control cows (p < 0.05). Retinol, certain metabolic parameters and metabolic hormones concentrations did not differ between beta-carotene and control cows. In addition, serum retinol concentration in beta-carotene cows without luteal activity was lower than in beta-carotene cows with luteal activity (p < 0.05), and serum gamma-glutamyl transpeptidase concentration in beta-carotene cows with luteal activity (p < 0.05) and control cows without luteal activity (p < 0.01) was higher than in control cows with luteal activity. In conclusion, beta carotene supply during the close-up dry period may support the onset of luteal activity during early lactation in dairy cows. PMID- 20002608 TI - Identification of two novel alleles HLA-B*070209 and HLA-B*130205 by polymerase chain reaction sequence-based typing. AB - HLA-B*070209 shows one nucleotide difference from B*070201 in exon 4. HLA B*130205 has a single nucleotide difference compared with HLA-B*130201. PMID- 20002609 TI - HLA DR-DQ combination associated with the increased risk of developing human HCV positive non-Hodgkin's lymphoma is related to the type II mixed cryoglobulinemia. AB - This investigation was focused on the contribution of individual human leukocyte antigen (HLA)-DR and -DQ alleles to the human hepatitis C virus (HCV)(+) non Hodgkin's lymphoma (NHL), with and without mixed cryoglobulinemia (MC), to study whether individual HLA class II alleles are expressed preferentially or equally in human HCV-specific NHL. For this purpose, peripheral blood mononuclear cells were obtained from two groups of patients with HCV(+) NHL and with or without MC (70 and 71 cases, respectively), and from 4575 blood donors. Eighty-three subjects with HCV infection only, and 118 patients with MC, only without lymphoma, were added as additional control groups. Individual HLA-DR and -DQ alleles were determined using high-resolution sequence-based typing and then data were collected by considering the HLA-DRB1 and DQB1 supertypes on the basis of common structural and functional features, proposed by in silico Bioinformatic studies. From the data, it is evidenced that the DR5-DQ3 HLA combination was strongly associated with the HCV (+) MC (+) NHL group of patients compared with bone marrow donor population (P or = 2 other risk factors. RESULTS: The counselling did not reduce the prevalence of overweight in 5- to 15-year-old participants. From age 7 onwards, the proportion of children with > or = 2 risk factors was lower in the intervention than in the control group (p = 0.005). At the age of 15 years, 13.0% of girls and 10.8% of boys in the intervention group and 17.5% of girls and 18.8% of boys in the control group had the risk factor cluster (p = 0.046 for main effect of the study group). Having even one risk factor at the age of 5 years predicted the clustering of risk factors at the age of 15 years (OR: 3.8, p < 0.001). CONCLUSION: Repeated, individualized dietary and lifestyle counselling may reduce the clustering of overweight-related cardiometabolic risk factors in adolescents even though the counselling is not intense enough to prevent overweight. PMID- 20002625 TI - Fusobacterium necrophorum multiple abscesses in children with sickle cell disease. PMID- 20002626 TI - Validity and reliability of a Swedish version of the Non-Communicating Children's Pain Checklist--Postoperative Version. AB - AIM: To test the validity and reliability of a Swedish version of the Non Communicating Children's Pain Checklist-Postoperative Version (NCCPC-PV). METHODS: Thirty-two consecutive children/adolescents (2-20 years of age) with cognitive impairment and no verbal communication from four habilitation centres were admitted to the study. Each child's behaviour was observed by a parent or a caregiver and by a physiotherapist in two calm and two painful situations within the child's everyday life. The raters independently assessed and graded the child's behaviour during 5 min according to the translated Swedish version of the NCCPC-PV. The intrarater and interrater reliability were determined, and the construct validity was examined. RESULTS: The results from 202 assessments showed that the construct validity was good: children's behavioural signs differed significantly between situations of pain and situations of calm (p < 0.001). Repeated assessments showed poor agreement both within and between raters [intraclass correlation coefficient (ICC) 0.51-0.65]. The agreement for pain was good (ICC 0.83). CONCLUSION: The Swedish version of the NCCPC-PV can be used for pain assessment in children with cognitive impairments who lack verbal communication. Aspects of reliability need to be further analysed. PMID- 20002627 TI - Interleukin-4 receptor polymorphisms in asthma and allergy: relation to different disease phenotypes. AB - AIM: Inheritance and genetic factors are supposed to influence susceptibility to asthma and allergy. We tested if single nucleotide polymorphisms (SNPs) in the IL4R gene were associated with susceptibility to such diseases, or if they were related to the phenotypic presentation of asthma and allergic rhinoconjunctivitis (ARC). METHODS: Three hundred and nine 12- to 13-year-old children were included. Six SNPs in the IL4R were analysed in response to current allergic disease, and to presentation of specific asthma and ARC phenotypes. Questionnaires were used to determine allergic disease status, and skin prick tests to evaluate sensitization to common airborne allergens. RESULTS: Less eczema was seen in individuals with the AA-genotype of rs2057768, and less ARC among those with the AA-genotype of rs2107356, especially ARC associated with sensitization to pollen. The AA-genotype of rs2057768 and the TT genotype of rs3024632 were associated with a specific asthma phenotype. CONCLUSION: Variations within the IL4R gene are associated with allergic diseases in children, preferably with eczema and disease phenotypes of ARC and asthma. PMID- 20002628 TI - Time for a change in the research paradigm for Alzheimer's disease: the value of a chaotic matrix modeling approach. AB - The amyloid cascade hypothesis, based on the genetic data from early onset, familial forms of the disease, has been the dominant model for many years and involves over production and deposition of the beta amyloid protein as causal in the disease process. However, it does not apply very well to the more common, later onset, sporadic form of the disease, where a wider range of factors appear to be involved in disease progression. Over recent years, data illustrating reciprocal interactions between the amyloid precursor protein (APP) and its various metabolites with many factors involved in normal synaptic plasticity have emerged. These feedback relationships have the potential to affect the complex kinase cascades involved in every aspect of neuronal function. Further, data regarding the multiple roles of the presenilins have the potential to allow the over expression and deposition of the amyloid beta protein to be both a cause and consequence of disease progression, with relevance in both sporadic and familial of Alzheimer's disease (AD). Disease progression might be better explained by a chaotic matrix of factors and raises the question again whether AD should be approached as a single entity or as a syndrome, with important consequences for disease identification and treatment. PMID- 20002629 TI - Impact of social and economic characteristics on orthodontic treatment among children and teenagers in France. AB - OBJECTIVES: This study aimed to estimate the prevalence of orthodontic treatment in France among children and teenagers aged 8-18 years, by sex and by age, and to investigate the specific role of social and economic characteristics on use of orthodontic treatment. METHODS: We analyzed data from the cross-sectional national health survey conducted in France in 2002-2003, which included a sample of 5988 children aged 8-18 years. All data were collected by interview including the question on orthodontic treatment. Other data used in our study were family social status and income, maternal educational attainment and place of birth, whether the child was covered by a supplementary health insurance and whether the residence was urban or rural. We also calculated the density of orthodontists in the district. Multivariate logistic regression analyses were used to study the relationships between these social and economic factors and orthodontic treatment. RESULTS: The prevalence of orthodontic treatment was 14% of all children aged 8-18, 15% for girls, and 13% for boys, and 23% in the 12 to 15-year age group. Children were less likely to have orthodontic treatment when parents were service or sales workers compared with children whose parents were managers or professionals (aOR = 0.50; 95%CI: [0.34;0.76]), when family income was in the lowest, compared with highest quartile (aOR = 0,62; 95% CI: [0.45;0.85]), when children had no supplementary insurance compared with children covered by private insurance (aOR = 0.53; 95% CI: [0.34; 0.81]), or when they lived in rural compared with urban areas (aOR = 0.70; 95% CI: [0.54; 0.91]). CONCLUSION: There are social inequalities in orthodontic treatment in France, associated mainly with social status, annual income, supplementary insurance, and the residence area. PMID- 20002630 TI - PUFA--an index of clinical consequences of untreated dental caries. AB - RATIONALE: Dental caries is a global public health problem, especially in children. Most caries in developing countries remains untreated. Only limited data are available on the clinical consequences of untreated dental caries because there is no measure to quantify the prevalence and severity of oral conditions resulting from untreated dental caries. OBJECTIVES: To present a new index to evaluate the prevalence and severity of oral conditions resulting from untreated dental caries. To validate the index within the Philippines National Oral Health Survey, 2006. METHODS: The PUFA index records the presence of severely decayed teeth with visible pulpal involvement (P/p), ulceration caused by dislocated tooth fragments (U/u), fistula (F/f) and abscess (A/a). RESULTS: Good kappa values show the reliability of the index. The prevalence of PUFA/pufa >0 was 85% and 56% for 6- and 12-year-olds, respectively. The mean number of teeth affected (PUFA/pufa) was 3.5 and 1.2 for 6- and 12-year-olds, respectively. In 6- and 12-year-olds, 40% and 41% of decayed teeth had progressed to odontogenic infections. CONCLUSION: The PUFA index complements classical caries indices with relevant information for epidemiologists and health care planners. PMID- 20002631 TI - A literature review of aesthetic perceptions of dental fluorosis and relationships with psychosocial aspects/oral health-related quality of life. AB - Aesthetic perceptions and oral health-related quality of life concerning dental fluorosis have been assessed in several studies during the past two decades. However, no comprehensive review article summarizing the studies investigating this issue has been published. OBJECTIVE: To assess the relationships between perceptions of dental appearance/oral health-related quality of life (OHRQoL) and dental fluorosis. METHODS: The PubMed database was searched using the Medical Subject Headings (MeSH) for English-language studies from 1985 to March 2009. Thirty-five articles qualified for inclusion and then were classified into three categories based on the type of study approach: (i) respondent review of photographs and assessment concerning satisfaction/acceptance, (ii) respondent assessment of study subject's teeth concerning satisfaction/acceptance, and (iii) respondent assessments of the psychosocial/OHRQoL impact. RESULTS: There were varied results from earlier studies focused on satisfaction/acceptance of very mild to mild fluorosis. More recent studies with methodological improvements to assess impact on quality of life clearly showed that mild fluorosis was not a concern. Furthermore, mild fluorosis was sometimes associated with improved OHRQoL. Severe fluorosis was consistently reported to have negative effects on OHRQoL. CONCLUSION: Because dental fluorosis in the United States and other nations without high levels of naturally-occurring fluoride is mild or very mild, with little impact on OHRQoL, dental professionals should emphasize the appropriate use of fluorides for caries prevention and preventing moderate/severe fluorosis. PMID- 20002632 TI - Rapid assessment and safe management of severe pulmonary hypertension with milrinone during orthotopic liver transplantation. AB - The incidence of porto-pulmonary hypertension (PPHN) in patients with end stage liver disease is 8.5%. Evidence indicates that proceeding with orthotopic liver transplantation (OLT) in patients diagnosed with severe PPHN (mean pulmonary artery pressure [mPAP]>45 mmHg) at the time of OLT surgery is associated with high perioperative mortality. We describe a case of severe PPHN that was diagnosed by right heart catheterization at the time of surgery. We quickly determined the reversibility of PPHN with a bolus of milrinone and proceeded with OLT. Further episodes of pulmonary hypertension were successfully managed with continuous milrinone infusion and transesophageal echocardiography monitoring. Reversibility via vasodilator trial after identification of high pulmonary artery pressures (PAP) may be an important indication of the feasibility of OLT. Milrinone may be useful for the rapid identification of the reversibility of high PAP and may be an effective agent to control abrupt increases in PAP during OLT. PMID- 20002633 TI - Syringocystadenocarcinoma papilliferum. PMID- 20002634 TI - Porokeratosis ptychotropica: successful treatment with the dermatome. PMID- 20002635 TI - A rare ischemic complication of ingrowing toenail surgery in a child. PMID- 20002636 TI - A rare case of a giant Marjolin ulcer. PMID- 20002637 TI - Botulinum toxin type A for the treatment of glabellar lines in Chinese: a double blind, randomized, placebo-controlled study. AB - BACKGROUND: This is the first well-controlled study of the use of botulinum toxin type A (BoNTA) for glabellar lines in China. OBJECTIVES: To evaluate the safety and efficacy of BoNTA in the treatment of glabellar lines in Chinese subjects. MATERIALS AND METHODS: A total of 227 subjects received a single treatment in a 3:1 randomization ratio of BoNTA (20 U):placebo and were observed for 120 days after injection. Effective outcome measures included investigator's rating of wrinkle severity at maximum frown and rest and subjects' global assessment and self-perception of age. RESULTS: A significantly higher responder rate at maximum frown, ranging from 94.1% at day 30 to 52.9% at day 120, was noted in the BoNTA group. The proportion of subjects with none or mild glabellar lines at rest was 66.7% in the BoNTA group at day 30. Most (95.3%) of the subjects treated with BoNTA reported better than 50% improvement at day 30, and self-perception of age was less than chronological age. There were no statistically significant differences in adverse events reported between the two groups (p=.06). CONCLUSION: A single treatment of 20 U of BoNTA was effective and safe in reducing glabellar lines in Chinese subjects. PMID- 20002638 TI - Reduced pain with use of proprietary hyaluronic acid with lidocaine for correction of nasolabial folds: a patient-blinded, prospective, randomized controlled trial. AB - BACKGROUND: Pain during and after implantation of dermal gel fillers is a consistent complaint of patients undergoing soft tissue augmentation. Reduction of pain during injection would increase patient comfort and improve the overall patient experience. OBJECTIVE: To evaluate pain at the injection site during and after the injection of Prevelle SILK or Captique and to evaluate outcomes after 2 weeks. METHODS & MATERIALS: In a patient-blinded, prospective, randomized, split face design trial, a non-animal-derived hyaluronic acid based filler formulated with lidocaine (Prevelle SILK) was injected in one nasolabial fold (NLF), and the same filler without lidocaine (Captique) was injected in the contralateral NLF of 45 enrolled patients. Injection site pain was measured using a visual analogue scale at injection (time 0) and 15, 30, 45, and 60 minutes after injection. Patients were asked to return for an evaluation after 2 weeks and to complete a self-assessment questionnaire during the follow-up visit. RESULTS: There was more than 50% less pain associated with the dermal gel with lidocaine than with the same filler without lidocaine at all time points (p<.05). The greatest difference in pain was recorded at the time of injection, and then the effect gradually declined over the 60-minute period. Both fillers were well tolerated, and there was no difference in outcome after 2 weeks. CONCLUSION: Addition of lidocaine to a filler resulted in significantly less pain associated with the procedure without compromising outcomes. PMID- 20002639 TI - Reconstruction of a helical rim defect. PMID- 20002640 TI - Columellar advancement flap for midline nasal defects. PMID- 20002641 TI - Surgical gem: inverted contralateral ear digital photograph as a template during earlobe keloid reconstruction. PMID- 20002642 TI - Needle preference in patients receiving cosmetic botulinum toxin type A. AB - BACKGROUND: Patients often complain of pain and bruising from needle injections. Some clinicians believe smaller gauge needles cause less pain. Thirty-gauge needles are currently the standard needles employed for administering botulinum toxin type A (BTX-A). OBJECTIVE: This study sought to determine whether patients receiving BTX-A have a preference for 30-gauge or 32-gauge needles based on the amount of pain and bruising experienced. METHODS: Thirty-seven subjects received BTX-A on the right side of the face using a 30-gauge needle and on the left side using a 32-gauge needle. Subjects were masked to needle size. They were then asked to rate injection pain on an 11-point numerical rating scale and to note any bruising. Physician preference was also evaluated. RESULTS: There were no statistically significant differences in the amount of intra-procedural pain (p=.37) or the level of post-procedural pain and discomfort (p=.76) experienced. Twenty-seven percent of subjects reported greater bruising with the 32-gauge needle, versus 29.7% with the 30-gauge needle. The physician injector did not have a preference. Lastly, 83.8% of subjects did not detect a difference in BTX-A paralysis effect. CONCLUSION: We do not recommend using 32-gauge needles in place of 30-gauge needles for administering BTX-A. PMID- 20002643 TI - Commentary: hair and pilonidal sinus disease. PMID- 20002644 TI - Evaluation of 60 patients with pilonidal sinus treated with laser epilation after surgery. AB - BACKGROUND: The surgical treatments for pilonidal sinus disease often result in recurrences, and the patients risk requiring multiple surgical interventions. OBJECTIVE: To evaluate the role of alexandrite laser hair removal after surgery in pilonidal sinus disease. METHODS: Sixty patients who underwent surgical treatment of pilonidal sinus disease and were treated with a 755-nm alexandrite laser after surgery between 1999 and 2007 were examined retrospectively. The charts were reviewed, and the patients were interviewed on the telephone about their post-laser period and recurrence. The laser parameters, patient history, and surgical details were recorded. RESULTS: The overall recurrence rate was 13.3%, after a mean follow-up period +/- standard error of the mean of 4.8 +/- 0.3 years. The mean number of laser treatment was 2.7 +/- 0.1. Seventy-five percent of the recurrences were detected after a follow-up period of 5 to 9 years. Fifty percent of the recurrent cases had drainage and secondary intention before the laser epilation. CONCLUSION: Our results strongly suggest that laser hair removal after surgical interventions in pilonidal sinus disease decreases the risk of recurrence over the long term. PMID- 20002645 TI - Commentary: the evolving role of photodynamic therapy for the treatment of photoaged skin. PMID- 20002646 TI - Photorejuvenation with topical methyl aminolevulinate and red light: a randomized, prospective, clinical, histopathologic, and morphometric study. AB - BACKGROUND: Photodynamic therapy (PDT) is an option for skin rejuvenation. Although many studies report clinical improvement with PDT in photodamaged skin, histologic and morphometric evidence is not documented in most cases. OBJECTIVE: To evaluate clinical and histopathologic changes induced by methyl aminolevulinate (MAL)-PDT and to morphometrically quantify collagen and elastic fibers in skin remodeling induced by MAL-PDT in photodamaged skin. METHODS AND MATERIALS: Fourteen patients were treated with two sessions of MAL-PDT. The light source was a light-emitting diode: 635 nm, 37 J/cm(2). Skin biopsies were performed before and 3 and 4 months after treatment. All fragments were stained using the hematoxylin-eosin, orcein, and picrosirius techniques. Morphometric studies were done of three samples from each patient. RESULTS: Global clinical improvement was observed in 10 of 14 patients. The histopathologic study showed increased collagen fibers 3 and 6 months after treatment. The decrease in the amount of elastic fiber was statistically significant 3 (p=.016) and 6 (p=.008) months after treatment. The increase in the amount of collagen fiber was statistically significant 6 months after treatment (p=.048). CONCLUSION: Clinical improvement with regard to texture, firmness, wrinkle depth, skin coloration, and clearance of actinic keratoses was observed. Histopathologic and morphometric studies were consistent with the clinical findings. PMID- 20002647 TI - Radiofrequency ablation of facial nerve branches controlling glabellar frowning. AB - BACKGROUND: Hyperdynamic activity of the corrugator supercilii and procerus muscles causes glabellar furrows. Recently, a novel radiofrequency device has become available that can effectively ablate the efferent nerves controlling corrugator and procerus contraction, producing clinical results that are similar to those of botulinum toxin. OBJECTIVE: To assess the efficacy, longevity of effect, and side effects of the radiofrequency ablation device in the treatment of hyperdynamic glabellar furrows. MATERIALS AND METHODS: Four probe entry points were used to access branches of the temporal and angular nerves. Seven and two ablations, respectively, were delivered to each temporal branch and angular nerve. RESULTS: Twenty-nine patients underwent bilateral radiofrequency ablation of temporal branches of the facial nerve and the angular nerves. Abrogation of glabellar furrowing was achieved in 90% of patients. No major adverse events were observed. All patients developed mild to moderate swelling, and nine patients (31%) developed purpura in the treated areas. Sixty-nine percent of patients had effects that lasted 4 months or longer, 41% had effects that lasted 6 months or longer, and 10% had effects lasting longer than 12 months. CONCLUSION: Radiofrequency ablation of efferent branches of the temporal and angular nerves effectively eliminates corrugator and procerus contraction and concomitant glabellar furrowing. PMID- 20002648 TI - Treatment needs in HIV prevention trials: using beneficence to clarify sponsor investigator responsibilities. AB - Some participants will get HIV-infected in HIV prevention trials, despite risk reduction measures. The subsequent treatment responsibilities of sponsor investigators have been widely debated, especially where access to antiretroviral therapy (ART) is not available. In this paper, we explore two accounts of beneficence to establish whether they can shed light on sponsor-investigator responsibilities. We find the notion of general beneficence helpful insofar as it clarifies that some beneficent actions will be obligatory where they can be dispensed without scuppering the trial. We find the notion of specific beneficence helpful insofar as it directs investigators to attend to the needs of trial participants; however the range of interventions that could be provided remains unhelpfully broad. We then examine accounts of the investigator participant relationship to narrow the range of interventions that investigators should provide, concluding that health-care, and HIV infection, are appropriate foci. We conclude that when investigators are able to meet the ART needs of their participants (e.g. referral, assisted referral or direct provision) without sacrificing trial quality, they must do so. However, there is little of this explicit direction to be found in the account of specific beneficence itself, but rather it is found in accounts of the relationship that are compatible with beneficence. PMID- 20002649 TI - Periadventitial delivery of anti-EGF receptor antibody inhibits neointimal macrophage accumulation after angioplasty in a hypercholesterolaemic rabbit. AB - Monocyte recruitment and their differentiation into macrophages are both early events in native and accelerated atherosclerosis that follows angioplasty. We have investigated the putative functional role of the epidermal growth factor receptor (EGFR) present on rabbit monocytes/macrophages. The impact of periadventitial delivery of an EGFR-specific, blocking monoclonal antibody (ICR62, which inhibits EGF-binding to its receptor) was investigated in a rabbit model of accelerated atherosclerosis induced by a combination of carotid injury and 4 weeks of a 2% cholesterol-diet. Two weeks after the initiation of the diet, a balloon-catheter angioplasty of the left common carotid artery was performed and a collar placed around the injured carotid artery immediately, for the delivery of ICR62 antibody, isotype-matched antibody or saline control. Monocyte/macrophage accumulation, cell proliferation and neointimal thickening were determined 2 weeks after the delivery of the antibodies. The function of the EGFR on rabbit monocytes was also investigated in vitro, using chemotaxis assays. Treatment with ICR62 was associated with a significant reduction in macrophage accumulation and neointimal thickening and a 76% reduction in neointimal area of the vessel wall compared with controls. In vitro ICR62 inhibited macrophage and smooth muscle cell migration towards EGFR ligands including EGF and HB-EGF. These findings suggest that EGFR ligation may be important in the development of early atherosclerotic lesions following balloon-catheter angioplasty, and periadventitial delivery may provide a feasible approach for administration of the inhibitors of EGFR-binding such as ICR62. PMID- 20002650 TI - Accurate identification of proliferative index in melanocytic neoplasms with Melan-A/Ki-67 double stain. PMID- 20002651 TI - Cutaneous T-cell lymphoma occurring with a melanocytic proliferation, masquerading as a nonhealing ulcer with reactive changes. AB - Two of the most challenging areas in dermatopathology are lymphoproliferative disorders and melanocytic lesions. We present a case of peripheral T-cell lymphoma occurring with an intradermal melanocytic proliferation. A 63-year-old Caucasian man presented with a 12-cm edematous, erythematous to violaceous, scalp ulceration that had enlarged over six months. Previous biopsies showed reactive changes which were concerning for infection. The last biopsies showed small to intermediate sized, angulated cells with clear cytoplasm within the dermis, with extension into the epidermis. These cells stained positive with markers for CD3, CD45RO and CD43, yet showed decreased expression of pan-T-cell markers CD5 and CD7, and absent expression of CD4, CD8, CD56 and CD57 and EBV. Molecular studies showed a clonal T-cell receptor gamma chain gene rearrangement. The diagnosis was peripheral T-cell lymphoma, unspecified. Another biopsy from an indurated area separate from the ulcer showed scattered, enlarged cells embedded in the same lymphocytic infiltrate. No mitotic figures were identified. These cells stained for S100 and Melan-A, in a partly nested arrangement. This was felt to represent a melanocytic nevus. This case likely represents an extraordinary coincidence of two distinctly different neoplasms. PMID- 20002652 TI - Candida balanitis: risk factors. AB - BACKGROUND: The amount of available information on the prevalence and incidence of candida balanitis is still surprisingly scarce. OBJECTIVES: To determine the prevalence of candida colonization and candida balanitis in men attending a Sexually Transmitted Diseases (STD) clinic. To identify risk factors associated with candida balanitis. METHODS: During a 36-month period, a cross-sectional study was carried out on consecutive men attendees of the STD clinic in Hospital S. Joao, Porto. Clinical and epidemiological data were recorded. Specimen collection from the glans penis and the coronal sulcus followed two procedures: a cotton tipped swab and the direct impression on the surface of CHROMagar Candida medium. Risk factors were considered singly and in combination through logistic regression models. RESULTS: Among 478 men enrolled, the prevalence of candida colonization was 26.2% and the prevalence of candida balanitis was 18%. Candida colonization was strongly associated with an age above 60 years (OR = 3.375; 95% CI: 1.547-7.362) and with the presence of other cause of balanitis apart from Candida organisms (OR: 2.466; 95% CI: 1.491-4.078). An age above 40 years (OR: 2.27; 95% CI: 1.005-4.500), diabetes mellitus (OR: 19.390; 95% CI: 7.789-48.273) and more than ten candida colonies recovered by culture (OR: 9.586; 95% CI: 2.682 34.263) were risk factors for candida balanitis. CONCLUSIONS: This study highlights the impact of factors other than sexual behaviours upon the epidemiology of this infection. For both candida colonization and infection, age was an important risk factor. Diabetes mellitus was an independent risk factor for candida balanitis. More than ten colonies recovered from culture are associated with clinical signs and symptoms. PMID- 20002653 TI - C-reactive protein and leucocyte activation in psoriasis vulgaris according to severity and therapy. AB - BACKGROUND: Psoriasis vulgaris is a chronic recurrent inflammatory skin disease and psoriatic lesions have shown leucocyte infiltration. OBJECTIVES: We aimed to study C-reactive protein (CRP) and leucocyte activation markers/inhibitors as potential monitors of psoriasis vulgaris. METHODS: A cross-sectional (n = 73) and a longitudinal study (before, at 3, 6 and 12 weeks of therapy; n = 47) was performed; 10 patients started topical treatment, 17 narrow-band ultraviolet light B (NBUVB) and 20 psolaren associated to UVA (PUVA); psoriasis severity was defined by Psoriasis Area and Severity Index (PASI). RESULTS: Compared with control (n = 38), we found higher CRP levels, total leukocyte/neutrophil count, elastase, lactoferrin and alpha1-antitrypsin. Increasing PASI was linked to increasing CRP and a trend to higher elastase and lactoferrin, suggesting that worsening enhances inflammatory response with neutrophil activation. CRP correlated with PASI, total leucocytes, neutrophils, elastase, lactoferrin and alpha1-antitrypsin. NBUVB and PUVA presented similar effects. CONCLUSION: We propose CRP as a useful marker of psoriasis severity that could be used to monitor psoriasis and its treatment, and, together with PASI and elastase, could also be used as a global index of severity. PMID- 20002654 TI - Intense pulsed light vs. pulsed-dye laser in the treatment of facial acne: a randomized split-face trial. AB - BACKGROUND: Various laser and light therapy have been increasingly used for the treatment of acne vulgaris. PATIENTS AND METHODS: Twenty patients with facial acne were treated using intense pulsed light (IPL) on one side of the face and pulsed dye laser (PDL) on the other to compare the efficacy and safety of IPL and PDL. Treatment was performed 4 times at 2-week intervals. Treatment effectiveness was determined using lesion counts, acne severity, patient subjective self assessments of improvement, and histopathological examinations, which included immunohistochemical staining for transforming growth factor-beta (TGF-beta). RESULTS: Numbers of total acne lesions decreased following both treatments. For inflammatory lesions such as papules, pustules and nodules, IPL-treated sides showed an earlier and more profound improvement than PDL-treated sides. However, at 8 weeks after the 4th treatment, a rebound aggravation of acne was observed on IPL-treated sides. On the contrary, PDL produced gradual improvements during the treatment sessions and these improvements lasted 8 weeks after the 4th treatment. Non-inflammatory lesions as open and closed comedones also showed improvement following both treatments and PDL-treated sides showed better improvement as the study proceeded. Histopathological examinations showed amelioration in inflammatory reactions and an increase in TGF-beta expression after both treatments, which were more prominent for PDL-treated sides. CONCLUSION: Both PDL and IPL were found to treat acne effectively, but PDL showed a more sustained effect. TGF-beta might play a key role in the resolution of inflammatory acne lesions. PMID- 20002655 TI - No interaction between methyl jasmonate and ozone in Pima cotton: growth and allocation respond independently to both. AB - Ozone (O3) is damaging to plants, inducing signalling pathways involving antagonism between jasmonates and ethylene. These pathways mediate O3 responses, particularly to acute exposure, and their manipulation protected several species against acute and chronic O3. We use chronic daily exposure of up to 163 ppb O3, and twice weekly application of up to 320 microg plant(-1) methyl jasmonate (MeJA) to test two hypothesizes: 1) a low rate of MeJA does not affect growth but increases O3 sensitivity; 2) a high rate inhibits growth but reduces O3 sensitivity. Both hypotheses were rejected. Growth declined with increases in both MeJA and O3. MeJA at 40 microg plant(-1) caused no direct effect, and at 160 microg plant(-1) reduced growth similarly at all O3. Neither rate altered O3 sensitivity. These additive responses are not consistent with protection by MeJA in this system. They may reflect inter-specific differences in signalling, since O3 concentrations used here exceeded some reported acute exposures. Alternatively, parallel responses to O3 and MeJA may suggest that O3-induced jasmonates play a developmental role in chronic response but no protective role in the absence of lesions characteristic of acute exposure. MeJA appears useful as a probe of these mechanisms. PMID- 20002656 TI - Structure and dynamics of a two-helix SNARE complex in live cells. AB - SNAREs are clustered membrane proteins essential for intracellular fusion steps. During fusion, three to four SNAREs with a Q(a)-, Q(b)-, Q(c)- and R-SNARE-motif form a complex. The core complex represents a Q(a)Q(b)Q(c)R-SNARE-motif bundle, most certainly assembling in steps. However, to date it is unknown which intermediate SNARE complex observed in vitro also exists in vivo. Here we have applied comparative fluorescence recovery after photobleaching (FRAP)-studies as a novel approach for studying in intact cells a SNARE interaction involved in synaptic vesicle fusion [catalyzed by syntaxin 1A (Q(a)), SNAP25 (Q(b)/Q(c)) and synaptobrevin 2 (R)]. We find that the Q(b)-SNARE-motif of SNAP25 interacts reversibly with clustered syntaxin. The interaction requires most of the alpha helical Q(b)-SNARE-motif and depends on its position within the molecule. We conclude that a zippered Q(a)Q(b)-SNARE complex represents a short-lived SNARE intermediate in intact cells, most likely providing an initial molecular platform toward membrane fusion. PMID- 20002657 TI - Liver transplantation from donation after cardiac death donors: initial Belgian experience 2003-2007. AB - The Belgian experience with donation after cardiac death (DCD) liver transplantation (LT) was retrospectively reviewed, particularly evaluating patient and graft survivals, and biliary complications. From 2003 to 2007, 58 DCD LT were performed in Belgium. Mean procurement total warm ischemia time was 25 +/ 2 min (mean +/- SEM). Mean cold ischemia time was 451 +/- 18 min. Mean follow-up was 23 +/- 2.2 months. Post-transplant peak aspartate aminotransminases was 2241 +/- 338 UI/l. Patient survivals at 1 month, 1 and 3 years, were 91.3%, 83.3% and 66.9% respectively. Graft survivals at 1 month, 1 and 3 years, were 84.4%, 72.4% and 48.8% respectively. Two patients (3.4%) developed primary nonfunction. Regarding the biliary complications, seven grafts (12%) were lost because of intrahepatic cholangiopathy, and 12 other patients (20.6%) developed bile duct stenoses requiring endoscopic and/or surgical management. The rate of symptomatic ischemic biliary lesions for grafts surviving more than 3 months was 38% (19/50). Although DCD organ donors may be a source of viable liver grafts, results were inferior to those obtained with donation after brain death LT in this series. Prognostic criteria have to be developed to improve results of DCD-LT. PMID- 20002658 TI - No evidence of occult hepatitis C virus (HCV) infection in serum of HCV antibody positive HCV RNA-negative kidney-transplant patients. AB - Persistence of hepatitis C virus (HCV) in patients who cleared HCV is still debated. Occult HCV infection is described as the presence of detectable HCV RNA in liver or peripheral blood mononuclear cells (PBMCs) of patients with undetectable plasma HCV-RNA by conventional PCR assays. We have assessed the persistence of HCV in 26 kidney-transplant patients, followed up for 10.5 years (range 2-16), after HCV elimination while on hemodialysis. If HCV really did persist, arising out of the loss of immune control caused by institution of the regimen of immunosuppressive drugs after kidney transplantation, HCV reactivation would have taken place. Their immunosuppression relied on calcineurin inhibitors (100%), and/or steroids (62%), and/or antimetabolites (94%). An induction therapy, given to 22 patients, relied on rabbit antithymocyte globulin (59%) or anti-IL2-receptor blockers (32%). All patients had undetectable HCV RNA as ascertained by several conventional tests. At the last follow-up, no residual HCV RNA was detected in the five liver biopsies, the 26 plasma, and in the 37 nonstimulated and 24 stimulated PBMCs tested with an ultrasensitive RT-PCR assay (detection limit, 2 IU/ml). No biochemical or virologic relapse was seen during follow-up. The absence of HCV relapse in formerly HCV-infected immunocompromised patients suggests the complete eradication of HCV after its elimination while on dialysis. PMID- 20002659 TI - Food allergy to meat and milk in adults. PMID- 20002660 TI - Influence of acupuncture on type I hypersensitivity itch and the wheal and flare response in adults with atopic eczema - a blinded, randomized, placebo controlled, crossover trial. AB - BACKGROUND: Itch is a major symptom of allergic skin disease. Acupuncture has been shown to exhibit a significant effect on histamine-induced itch in healthy volunteers. We investigated the effect of acupuncture on type I hypersensitivity itch and skin reaction in a double-blind, randomized, placebo-controlled, crossover trial. METHODS: An allergen stimulus (house dust mite or grass pollen skin prick) was applied to 30 patients with atopic eczema before (direct effect) and after (preventive effect) two experimental approaches or control observation: acupuncture at points Quchi and Xuehai [verum acupuncture (VA), dominant side], 'placebo-point' acupuncture (PA, dominant side), no acupuncture (NA). Itch intensity was recorded on a visual analogue scale. After 10 min, wheal and flare size and skin perfusion (via LASER-Doppler) were measured at the stimulus site, and the validated Eppendorf Itch Questionnaire (EIQ) was answered. RESULTS: Mean itch intensity was significantly lower in VA (35.7 +/- 6.4) compared to NA (45.9 +/- 7.8) and PA (40.4 +/- 5.8) regarding the direct effect; and significantly lower in VA (34.3 +/- 7.1) and PA (37.8 +/- 5.6) compared to NA (44.6 +/- 6.2) regarding the preventive effect. In the preventive approach, mean wheal and flare size were significantly smaller in VA (0.38 +/- 0.12 cm(2)/8.1 +/- 2.0 cm(2)) compared to PA (0.54 +/- 0.13 cm(2)/13.5 +/- 2.8 cm(2)) and NA (0.73 +/- 0.28 cm(2)/15.1 +/- 4.1 cm(2)), and mean perfusion in VA (72.4 +/- 10.7) compared to NA (84.1 +/- 10.7). Mean EIQ ratings were significantly lower in VA compared to NA and PA in the treatment approach; and significantly lower in VA and PA compared to NA in the preventive approach. CONCLUSIONS: Acupuncture at the correct points showed a significant reduction in type I hypersensitivity itch in patients with atopic eczema. With time the preventive point-specific effect diminished with regard to subjective itch sensation, whereas it increased in suppressing skin-prick reactions. PMID- 20002662 TI - IgE-mediated anaphylaxis to Hippobosca equina in a patient with systemic mastocytosis. PMID- 20002661 TI - Markers of eosinophilic and neutrophilic inflammation in bronchoalveolar lavage of asthmatic and atopic children. AB - BACKGROUND: Recent studies performing fiberoptic bronchoscopy in children have improved our understanding of asthma pathophysiology. Eosinophilic, but also neutrophilic, inflammation has been described in asthma, but the relationship with atopy was incompletely investigated. The aim of this study is to examine inflammatory cells and mediators in children with asthma compared to the appropriate controls, i.e. atopic children without asthma and children with no atopy or asthma. Moreover, asthmatic children were analysed separately based on the presence of atopy and stratified by age. METHODS: We recruited 191 children undergoing fiberoptic bronchoscopy for appropriate indications: 91 asthmatics (aged 1.4-17 years), 44 atopics without asthma (1.6-17.8 years) and 56 nonasthmatic nonatopic controls (1.4-14 years). In bronchoalveolar lavage, total and differential cell counts and inflammatory mediators, including ECP, eotaxin, IL-8 and TNFalpha, were analysed. RESULTS: Eosinophils and ECP levels were increased in asthmatic children when compared to controls (P = 0.002 and P = 0.01, respectively), but also atopic children without asthma had increased ECP levels compared to controls (P = 0.0001). Among asthmatic children, eosinophils and ECP levels were not different between atopic and nonatopic individuals. Neither neutrophils nor the related mediators (IL-8 and TNFalpha) differed significantly in the three groups. This pattern of inflammation was observed in both preschool and school-aged asthmatic children. CONCLUSIONS: This study suggests that markers of eosinophilic, but not neutrophilic inflammation, are increased in asthmatic children and also in atopic children without asthma. Of interest, in asthmatic children, the activation of the eosinophilic response is not solely because of the presence of atopy. PMID- 20002663 TI - Renal medullary carcinoma: molecular, pathological and clinical evidence for treatment with topoisomerase-inhibiting therapy. AB - STUDY TYPE: Aetiology (case series) Level of Evidence 4. OBJECTIVE: To present the molecular rationale and potential clinical benefit of topoisomerase II (TopoII)-inhibiting therapy for renal medullary carcinoma (RMC), a rare but extremely lethal form of kidney cancer that classically afflicts young men with sickle-cell trait. The current therapeutic approach with these aggressive tumours is radical nephrectomy followed by systemic chemotherapy, but the prognosis remains dismal. MATERIALS AND METHODS: The whole-genome expression was analysed in four RMC tumours. We also report a case of metastatic RMC in which a complete response was achieved for 9 months using a TopoII-inhibiting therapy. RESULTS: Expanded whole-genome expression analysis showed increases of TopoII in all cases. There was also overall deregulation of DNA remodelling and repair, and an ontological association between RMC and urothelial carcinoma. Using a TopoII inhibiting agent, there was a complete response for 9 months in a patient with metastatic RMC. CONCLUSION: This report provides molecular evidence for the rational use of TopoII inhibitors in the treatment of RMC. PMID- 20002664 TI - Predicting individual outcomes after radical cystectomy: an external validation of current nomograms. AB - OBJECTIVE: To determine whether published nomograms, i.e. The International Bladder Cancer Nomogram Consortium (IBCNC) and the Bladder Cancer Research Consortium (BCRC) statistical models for predicting disease recurrence and survival of patients after radical cystectomy, are feasible for routine use in intermediate-volume institutions in Europe, as although these nomograms had high accuracy by internal validation tests, they stem from high-volume centres and have not been validated elsewhere and thus their general applicability remains unproven. PATIENTS AND METHODS: We externally validated the published nomograms. Information about treatments, pathological details, and recurrence and survival status was retrospectively collected from 246 patients. The expected survival according to the nomograms was calculated. The predictive accuracy of the proposed models was calculated by Harrell's concordance indices. To assess the independent prognostic value of the variables proposed by IBCNC and BCRC, stepwise multivariable Cox regressions based on Akaike's Information Criterion (AIC) for the different endpoints were used. A best model for prediction was created on the basis of our data. RESULTS: The IBCNC and the BCRC nomograms showed an improvement in the predictive accuracy of recurrence, all-cause and bladder-cancer-specific survival after radical cystectomy of up to 4% compared to Tumour-Node-Metastasis stage-based predictions. According to the calculated AIC values for the different models, all nomograms performed better than the TNM classification. CONCLUSIONS: The BCRC and IBCNC nomograms provided accurate predictions when they were applied to an external cohort of patients from low- to intermediate-volume centres. The prediction of recurrence and survival based on the addressed nomograms is better than TNM stage-based prediction. The application of such nomograms can be supported on a sound basis, but further amendments are warranted. PMID- 20002665 TI - The effect of gender on response to bacillus Calmette-Guerin therapy for patients with non-muscle-invasive urothelial carcinoma of the bladder. AB - OBJECTIVE: To determine the influence of gender on the outcome of patients with high-risk non-muscle-invasive bladder cancer treated with intravesical bacille Calmette-Guerin (BCG) therapy, as the role of hormone status in the pathogenesis of urothelial carcinoma and the response to treatment remains subject to debate. PATIENTS AND METHODS: We reviewed 1021 consecutive patients (756 men and 265 women) who were treated with induction BCG between 1978 and 2006 for multiple or recurrent high-grade Ta, T1, and/or carcinoma in situ (CIS) bladder cancer. All patients had > or = 5 years of follow-up. The endpoints of initial response to BCG and the time to disease recurrence and progression were correlated with gender using Kaplan-Meier methods and multivariate Cox regression models. RESULTS: Men were significantly more likely to present with high grade (P = 0.003) tumours and with CIS (P < 0.001), while age and clinical stage at presentation were similar between men and women. There was no significant difference in the initial response to BCG by gender, as 593/756 (78.4%) men and 219/265 (82.6%) women had no evidence of disease at 6 months after BCG treatment (P = 0.14). The median time to recurrence after BCG therapy was also similar for men and women (20 vs 21 months, P = 0.51). Likewise, there was no evidence of a significant association between gender and the risk of disease progression after BCG therapy, such that the 5-year estimated freedom from progression was 77% and 82%, respectively, for men and women (P = 0.08). Moreover, on a multivariate analysis controlling for patient age and tumour stage, grade and CIS, gender was not associated with the risk of recurrence (hazard ratio 0.94, 95% confidence interval 0.79-1.11; P = 0.44) or progression (1.18, 0.85-1.63; P = 0.33) after BCG. When the outcomes for women treated with BCG were stratified by age <50 years (the median age of menopause in the USA) vs > or = 50 years, again there were no differences in the risk of tumour recurrence (P = 0.95) or progression (P = 0.35). CONCLUSION: These data suggest that the outcomes of men and women with high risk non-muscle-invasive urothelial carcinoma treated with BCG are similar. As such, further studies are required to determine the clinical relevance of preclinical evidence that has suggested a potential role for sex steroids in the pathophysiology of bladder cancer. PMID- 20002666 TI - Treatment failure and clinical progression after salvage therapy in men with biochemical recurrence after radical prostatectomy: radiotherapy vs androgen deprivation. AB - OBJECTIVE: To compare the outcomes between salvage radiotherapy (RT) and androgen deprivation therapy (ADT), to investigate factors determining clinical progression (CP) in men with prostate cancer. PATIENTS AND METHODS: The study comprised 121 patients with biochemical recurrence while on follow-up by prostate specific antigen (PSA) measurement, without adjuvant therapy after radical prostatectomy, received RT (45) or ADT (76). Failure after salvage therapy was defined as a PSA level of >0.2 ng/mL. Clinical, pathological and treatment factors were analysed. RESULTS: The clinicopathological characteristics were similar between the RT and ADT groups except that men in the RT group were younger (61.4 vs 65.4 years). After ADT, salvage failed in 10 (13%) after a mean (sd) of 18.5 (4.5) months of treatment, and 6.7 months after salvage failed all patients progressed clinically. After RT, salvage failed in 22 (49%) after 30.7 (5.2) months of response. Upon RT failure, all patients received ADT, after which in three (14%) patients the treatment failed again after 20.1 months of treatment and progressed to CP after 6.5 months, while in the remaining 19 (86%) patients the PSA level remained undetectable for 37.6 (7.7) months. On multivariate analysis, pathological stage (> or =T3b) and Gleason grade 5 disease were independently prognostic of CP. CONCLUSION: Salvage RT alone and combined with subsequent ADT provided PSA control in most patients, significantly increasing CP free survival compared with initial ADT. Patients with a short PSA doubling time (<3 months) are at high risk of failed salvage treatment after RT, and initial ADT might be considered. Regardless of salvage method, advanced pathological stage and Gleason grade 5 were factors prognostic of CP. PMID- 20002667 TI - Long-term data on the survival of patients with prostate cancer treated with radical prostatectomy in the prostate-specific antigen era. AB - STUDY TYPE: Therapy (case series) Level of Evidence 4. OBJECTIVE: To examine the long-term rates of biochemical recurrence (BCR)-free survival, cancer-specific mortality (CSM)-free survival, and overall survival (OS) in patients with prostate cancer treated with open radical prostatectomy (RP) in the prostate specific antigen (PSA) era. PATIENTS AND METHODS: The study comprised 436 patients who were treated with RP between 1992 and 1997 at our institution. None received adjuvant/salvage therapy in the absence of BCR. The BCR-free, CSM-free and OS rates were defined using the Kaplan-Meier method. Multivariable Cox regression models were used to test the effect of age, preoperative PSA level, neoadjuvant hormonal therapy, pT stage, lymph node status, RP Gleason sum and surgical margin status on BCR. RESULTS: The median follow-up of censored patients was 122, 128, and 132 months for, respectively, BCR-free, CSM-free and OS estimates. The 10-year event-free survival rates for the same endpoints were 60%, 94% and 86%, respectively. Preoperative PSA level, RP Gleason sum, pT stage, lymph node status, and surgical margin status were independent predictors of BCR (all adjusted P < 0.05). CONCLUSIONS: This study is the first to evaluate the long-term cancer control outcomes after RP from a European country in the PSA era. Our data indicate that RP provides excellent long-term survival rates in patients with clinically localized prostate cancer. Although approximately 40% of patients have BCR after 10 years of follow-up, the CSM rate after 10 years is as low as 6%. PMID- 20002668 TI - Efficacy, tolerability and safety of propiverine hydrochloride in comparison to oxybutynin in children with urge incontinence due to overactive bladder: Results of a multicentre observational cohort study. AB - OBJECTIVE: To compare, in a retrospective observational cohort study, the efficacy, tolerability and safety of propiverine and oxybutynin in children with urge incontinence (UI) due to overactive bladder. PATIENTS AND METHODS: Medical records were scrutinized for children with UI. As a primary efficacy outcome variable the achievement of continence after treatment with variable doses of propiverine or oxybutynin was assessed. Weekly UI episodes and daily voiding frequency were evaluated as secondary efficacy outcomes. Tolerability was evaluated by the rate of adverse events, adverse drug reactions caused by antimuscarinics and premature treatment termination. RESULTS: At 16 study centres, 621 children aged 5-14 years with UI due to overactive bladder were enrolled. After anticholinergic treatment (437 propiverine, 184 oxybutynin) continence was achieved in 61.6% and 58.7% of the patients after 186 and 259 days, respectively. There were clinically relevant improvements in voiding frequency across treatment groups. Daily doses of propiverine were markedly below the recommendations (0.54 vs 0.8 mg/kg body weight), daily doses of oxybutynin were according to the recommendations (0.31 vs 0.2-0.4 mg/kg body weight) at treatment initiation. There was a significantly more favourable tolerability to propiverine than oxybutynin for the overall rate of adverse events (3.9% vs 16.3%, odds ratio 4.813), adverse drug reactions caused by propiverine or oxybutynin (2.8% vs 9.2%) and premature treatment termination due to adverse drug reactions (1.6% vs 4.4%). CONCLUSION: Propiverine and oxybutynin are effective in children with UI due to overactive bladder. Sufficient treatment periods of at least 2, preferably 3-4, months are the crucial factors for a successful treatment. The tolerability profile of propiverine is better than for oxybutynin. PMID- 20002669 TI - Narrow-band imaging cystoscopy to evaluate bladder tumours--individual surgeon variability. AB - STUDY TYPE: Diagnosis (exploratory cohort) Level of Evidence 2b. OBJECTIVE: To assess individual urologist variability using narrow-band imaging (NBI) cystoscopy to evaluate bladder tumours. PATIENTS AND METHODS: In all, 50 patients underwent white-light and NBI cystoscopy to evaluate for recurrent bladder tumours. Endoscopic images in each patient were independently viewed by four urologists assessing presence or absence of tumour. Their findings were correlated with biopsy results. RESULTS: In all, 26 patients had recurrent tumour and 24 had benign histology. There were no significant differences among urologists detecting recurrent tumour or in determining final pathology. CONCLUSIONS: There does not appear to be a 'learning curve' for adapting to NBI surveillance cystoscopy in patients with bladder cancer. PMID- 20002670 TI - Lymph node mapping in patients with bladder cancer undergoing radical cystectomy and lymph node dissection to the level of the inferior mesenteric artery. AB - OBJECTIVE: To evaluate extended lymph node dissection (LND) as a nodal staging tool in the treatment of invasive carcinoma of the urinary bladder and to suggest a reasonable proximal limit of the dissection. PATIENTS AND METHODS: In all, 170 patients underwent radical cystectomy with extended LND up to the level of the inferior mesenteric artery. Specimens were evaluated as 13 separate packages from pre-designated anatomical locations. The number of LNs and presence of positive LNs (LN+) at each location was prospectively registered. RESULTS: The median (range) number of LNs removed was 24 (6-62). In all, 25.3% of the patients had LN+. The median (range) number of LN+ was 2 (1-20). Advanced T-stage was correlated with a higher risk of LN+ but not to the specific location of the LN+. Two patients had LN+ above the common iliac bifurcation with no LN+ more distally located within the pelvic region. All other patients with LN+ above the common iliac bifurcation had more distally located LN+. There were no skip lesions to LNs above the aortic bifurcation. CONCLUSIONS: Extended LND above the common iliac bifurcation including the presacral area provides a more accurate LN staging compared with a standard pelvic LND. Extending the limits above the aortic bifurcation is not necessary from a staging perspective. PMID- 20002671 TI - Extraprostatic extension of prostatic adenocarcinoma on needle core biopsy: report of 72 cases with clinical follow-up. AB - OBJECTIVE: To describe the histological findings and prognosis that are associated with extraprostatic extension (EPE) on needle core biopsy of prostatic adenocarcinoma. PATIENTS AND METHODS: We retrieved 99 cases of prostatic adenocarcinoma with EPE at initial diagnosis on biopsy from the consultation files of one of the authors between 1997 and 2009. The 72 cases that had available clinical follow-up data formed the basis of this study. RESULTS: The mean (range) age of the patients was 64 (48-87) years, the median (mean, range) serum prostatic specific antigen level was 7.8 (64.8, 0.3-1505) ng/mL, and 60 of the patients (83%) had abnormalities on a digital rectal examination. The mean (range) number of malignant cores was 7.7 (1-23); the mean percentage of carcinoma in each core was 69.6%, and that in the core(s) with EPE was 76.8%. The mean Gleason score in the core(s) with EPE was 8, with a mean highest Gleason score per case of 8.4. Perineural invasion was detected in 54 cases (75%). Ten of 11 patients treated surgically had EPE on the radical prostatectomy (RP) specimen; also six had positive resection margins, five showed invasion into the seminal vesicles and one had lymph node metastasis. The Gleason scores in nine of the RP specimens did not differ from the highest grade found in the associated biopsies (score 9 in three, 8 in two, 7 in four); in one case it increased (from score 6 to 8) and in one it decreased (from score 9 to 8). Patients were followed for a mean (median, range) of 2.9 (2, 0.1-9) years, with metastases identified in 29 (40%); 10 (14%) died from the disease. CONCLUSION: EPE on needle core biopsy of the prostate is strongly associated with extensive, high-grade prostatic adenocarcinoma, such that its usefulness as an isolated prognostic factor is relatively limited. PMID- 20002672 TI - Is it time to re-design the haematuria clinic? PMID- 20002673 TI - Chronic partial bladder outlet obstruction does not impair erectile function in male rats. AB - OBJECTIVE: To evaluate, in a well-controlled study, the effect of surgically induced partial bladder outlet obstruction (PBOO) on male erectile function in a rat model. MATERIALS AND METHODS: PBOO was created in 17 adult male Sprague Dawley rats by partial ligation of the proximal urethra. Sham-operated and PBOO rats were evaluated for urodynamic and erectile function at 4-8 weeks after surgery. Erectile responses to electrical field stimulation (EFS) to the major pelvic ganglion, and to erectogenic agents (1,1-diethyl-2-hydroxy-2-nitroso hydrazine, DEA-NO, and Y-27632) were evaluated and the area under the curve (AUC, a product of the intracavernous pressure and duration) was used to denote the erectile response. RESULTS: Experimental PBOO in rats significantly increased the mean (sem) bladder weight, to 256 (25) mg in PBOO rats vs 123 (24) mg in sham controls, and the voiding frequency to 1.01 (0.1) voids/min vs 0.72 (0.14) voids/min in sham controls (P < 0.05). There was no significant difference between the erectile response to EFS, with a mean AUC in sham control rats at 1.5, 3.0 and 4.5 V of 2603 (372), 3200 (332) and 3357 (166), respectively, vs 2273 (183), 3794 (211) and 4177 (306) in PBOO rats (P > 0.05); or to the erectogenic agents, the AUC for DEA-NO being 9000 (975) in PBOO rats vs 13 201 (2756) in sham controls, and the AUC for Y-27 632 being 44 915 (2462) and 45 907 (7408), respectively (P > 0.05). There was greater immunoreactivity to RhoA in bladder and penile tissues of PBOO than control rats. CONCLUSION: PBOO does not affect erectile function in rats. Additional mechanisms or pathways might be involved in lower urinary tract symptom-related erectile dysfunction in humans. PMID- 20002674 TI - Chronic inhibition of nitric-oxide synthase induces hypertension and erectile dysfunction in the rat that is not reversed by sildenafil. AB - STUDY TYPE: Aetiology (case control) Level of Evidence 3b OBJECTIVE To evaluate the effect of N(G)-nitro-L-arginine methyl ester (L-NAME)-induced hypertension (HT) on erectile function in the rat and determine if the phosphodiesterase (PDE) 5 inhibitor, sildenafil, can reverse the effects of nitric oxide (NO) deficiency, as HT is a risk factor for erectile dysfunction (ED) and the NO synthase (NOS) inhibitor L-NAME induces NO-deficient HT. MATERIALS AND METHODS: Thirty-six adult Sprague-Dawley male rats were divided into three groups, i.e. a control, L-NAME HT (40 mg/rat/day in the drinking water for 4 weeks), and sildenafil-treated L NAME-HT (1.5 mg/rat/day sildenafil, by oral gavage concomitantly with L-NAME). The erectile response expressed as a ratio of intracavernosal pressure (ICP)/mean arterial pressure (MAP), evaluated after electrical stimulation of the right cavernous nerve. The isometric tension of corpus cavernosum smooth muscle (CCSM) was measured in organ-bath experiments. NOS expression was determined immunohistochemically for neuronal (n)NOS and by Western blot analysis for endothelial (e) and inducible (i) NOS protein. cGMP levels were evaluated by enzyme-linked immunosorbent assay. RESULTS: The erectile response was diminished in the HT group. Nitrergic and endothelium-dependent relaxation was reduced, while the relaxation response to sodium nitroprusside and contractile response to phenylephrine were not altered in CCSM from L-NAME-treated rats. HT rats showed decreased expression of nNOS, whereas eNOS and iNOS protein expression was increased. Sildenafil partly restored endothelial and molecular changes in CCSM from HT rats, but did not reverse the decreased erectile response, even as cGMP levels returned to normal levels. CONCLUSIONS: Sildenafil treatment did not correct the ED in L-NAME-treated HT rats. Under sustained high blood pressure, up regulation of PDE5 expression failed to reverse the depletion of neuronal NO and/or impaired nNOS activity. However, endothelium-dependent relaxation was restored. Drug targeting of neuronal dysfunction might delay the onset of ED in HT. PMID- 20002675 TI - Syndecan-1 expression in prostate cancer and its value as biomarker for disease progression. AB - OBJECTIVE: To evaluate the association between syndecan-1 (CD138) expression and prostate cancer. PATIENTS AND METHODS: We evaluated syndecan-1 expression using a recently constructed tissue microarray of prostatic samples taken from 243 patients, corresponding to 1400 cores, with 69.8%, 5.6%, 17.6% and 7% of the cores representing localized prostate cancer, high-grade prostatic intraepithelial neoplasia, benign prostate tissue and hormone refractory/metastatic disease, respectively. RESULTS: Metastatic cases had the highest frequency and membranous staining intensity for syndecan-1 overexpression, followed by hormone refractory and localized disease (83.3% vs 34.8% and 25.7%, respectively). There was no significant difference in the frequency of membranous syndecan-1 expression between localized prostate cancer and benign glands (25.7% vs 24.7% of cases, respectively). However, benign glands showed significantly higher intensity staining than localized prostate cancer. We found no significant association between syndecan-1 expression and any of the following: Gleason score, pathological stage, surgical margin status and biochemical recurrence. CONCLUSION: The current available evidence, from the present and previous studies, show that syndecan-1 is not an independent predictor of recurrence or tumour-specific survival, diminishing its significance as a clinical marker. PMID- 20002676 TI - Management of urethral recurrence after orthotopic urinary diversion. AB - STUDY TYPE: Therapy (case series) Level of Evidence 4 OBJECTIVE To evaluate our experience with urethral recurrences in patients treated by radical cystectomy(RC) and orthotopic neobladder urinary diversion for carcinoma of the bladder. PATIENTS AND METHODS: We retrospectively reviewed the records of patients treated with RC and orthotopic urinary diversion between January 1980 and July 2004. RESULTS: In all, 260 patients underwent RC with a Studer or Hautmann orthotopic urinary diversion; the median (range) follow-up was 5.1 (0 15.6) years. Six patients (2.3%) developed local recurrence of urothelial cancer (UC) within the urethra after this treatment. The median (range) time to presentation with recurrence after RC was 2.4 (0.7-3.6) years for pT1-4 UC. Recurrences were treated with various methods, including transurethral resection, urethrectomy with conversion of neobladder to continent catheterizable diversion, and chemotherapy. At the last follow-up, four of these six patients were alive without disease, one was alive with disease, and one had died from disease. CONCLUSIONS: In our experience, local recurrences involving the urethra are infrequent. Complete surgical excision can provide a good outcome. Neoadjuvant chemotherapy should be considered for recurrences with adverse clinicopathological features. PMID- 20002677 TI - Comparative effects of T-type and L-type Ca(2+)-antagonists against noradrenaline induced contractions of human vas deferens. AB - OBJECTIVE: To investigate the effects of the relatively selective T-type Ca(2+) antagonists, mibefradil and flunarizine, and the L-type Ca(2+)-antagonist, nifedipine, on the contractions of longitudinal and circular muscles of human vas deferens, to elucidate the possible involvement of T-type voltage-gated Ca(2+) channels (VOCs) in the contractile function of the tissue. MATERIALS AND METHODS: Human vas deferens specimens from elective vasectomies were cut into strips of longitudinal muscle or transversely into rings of circular muscle. These were set up for tension recording and superfused with Krebs' medium (36 degrees C). Contractions were evoked by noradrenaline or high [K(+)](o) (in the presence of the L-type Ca(2+) agonist, FPL 64176) and the effects of Ca(2+) antagonists were determined. RESULTS: Noradrenaline (0.1-100 micromol/L) evoked rhythmic and tonic contractions of longitudinal and circular muscles, which were potently inhibited by nifedipine ( or = 3 months after vasectomy. PATIENTS AND METHODS: Between 1 July 2005 and 31 March 2008, 1073 men provided a first semen sample at > or = 3 months after vasectomy. Semen was first evaluated on a wet-slide preparation. Those samples with no ('azoospermia') or sporadic immotile spermatozoa could be cleared without further analysis. Samples with motile sperm were immediately labelled as potentially fertile, while those with a significant number of immotile sperm were re-analysed using a Neubauer haemocytometer. All samples with <100 000 immotile sperm/mL were cleared. RESULTS: Of men providing semen at 3 months after vasectomy, 96% could be cleared. No sperm were seen ('azoospermia') in 51.3% of samples, and 44.7% of samples contained <100 000 immotile sperm. No paternity has been reported in the cleared group after a follow-up of at least 1 year. CONCLUSIONS: A protocol stipulating that patients can be cleared after a single semen sample containing <100 000 immotile sperm/mL at > or = 3 months after vasectomy is safe and dramatically reduces the number of men who cannot be cleared at 3 months after vasectomy. PMID- 20002680 TI - (-)-Epigallocatechin-3-gallate induces Du145 prostate cancer cell death via downregulation of inhibitor of DNA binding 2, a dominant negative helix-loop helix protein. AB - (-)-Epigallocatechin-3-gallate (EGCG) is one of the major polyphenol components in green tea. It effectively induces apoptosis in prostate cancer cells. The anticancer effect of this reagent is appealing because it is a natural component of a popular daily beverage that has proven harmless for thousands of years, making it a good candidate chemopreventive agent. EGCG suppresses cell growth and causes cell death, but the mechanisms are not well characterized, especially in androgen-independent prostate cancer cells. In the present study, using Affymetrix genechip Hu133 2.0, we analyzed the gene expression patterns of the androgen-independent prostate cancer cell line Du145, treated with or without EGCG, and found 40 genes whose expression levels were altered (>twofold, either upregulated or downregulated, P < 0.01) upon treatment with EGCG. These gene products are involved in the functions of transcription, RNA processing, protein folding, phosphorylation, protein degradation, cell motility, and ion transport. Among them, inhibitor of DNA binding 2 (ID2), known as a dominant anti retinoblastoma (Rb) helix-loop-helix protein, was found to be downregulated fourfold by EGCG treatment. Forced expression of ID2 in Du145 cells reduced apoptosis and increased cell survival in the presence of EGCG, and knockdown ID2 expression in Du145 cells using a morpholino oligonucleotide specific for ID2 mimicked the apoptosis effect generated by EGCG treatment, although it was milder. To our knowledge, this is the first report indicating that ID2 is one of the critical factors in the signaling pathway of Du145 cell death induced by EGCG. PMID- 20002681 TI - Methylenetetrahydrofolate reductase polymorphisms, serum methylenetetrahydrofolate reductase levels, and risk of childhood acute lymphoblastic leukemia in a Chinese population. AB - Methylenetetrahydrofolate reductase (MTHFR), involved in DNA methylation and nucleotide synthesis, is thought to be associated with a decreased risk of adult and childhood acute lymphoblastic leukemia (ALL). Accumulating evidence has indicated that two common genetic variants, C677T and A1298C, are associated with cancer risk. We hypothesized that these two variants were associated with childhood ALL susceptibility and influence serum MTHFR levels. We genotyped these two polymorphisms and detected MTHFR levels in a case-control study of 361 cases and 508 controls. Compared with the 677CC and 677CC/CT genotypes, the 677TT genotype was associated with a statistically significantly decreased risk of childhood ALL (odds ratio = 0.53, 95% confidence interval = 0.32-0.88, and odds ratio = 0.55, 95% confidence interval = 0.35-0.88, respectively). In addition, a pronounced reduced risk of ALL was observed among low-risk ALL and B-phenotype ALL. Moreover, the mean serum MTHFR level was 8.01 ng/mL (+/-4.38) in cases and 9.27 ng/mL (+/-4.80) in controls (P < 0.001). MTHFR levels in subjects with 677TT genotype was significantly higher than those with 677CC genotype (P = 0.010) or 677CT genotype (P = 0.043) in controls. In conclusion, our results provide evidence that the MTHFR polymorphisms might contribute to reduced childhood ALL risk in this population. PMID- 20002682 TI - Transmission of periodontopathic bacteria from natural teeth to implants. AB - PURPOSE: Prevention of peri-implantitis is essential for the success of implant rehabilitation. Infection by periodontopathic bacteria is a major cause of peri implantitis. The aim of the present study was to identify the source of peri implant colonization by periodontopathic bacteria. MATERIALS AND METHODS: Twenty one patients with implants were enrolled in the study. Subgingival plaque samples from the adjacent, occluding, and contralateral natural teeth were collected prior to second-stage surgery. Samples from implant sulci were then obtained 2 weeks later. Detection of periodontopathic bacteria was performed by the polymerase chain reaction. RESULTS: The detection rates for Aggregatibacter actinomycetemcomitans, Prevotella intermedia, Porphyromonas gingivalis, Treponema denticola, Tannerella forsythia, and Fusobacterium nucleatum in all subgingival samples from natural teeth were similar to that in the peri-implant sulci. Multiple logistic regression analysis revealed an association between the detection of A. actinomycetemcomitans, Prevotella intermedia, Porphyromonas gingivalis, Treponema denticola, and F. nucleatum in the gingival crevices of adjacent teeth and that of the peri-implant sulcus, but no association for Tannerella forsythia. CONCLUSIONS: The present findings suggest that colonization by A. actinomycetemcomitans, Prevotella intermedia, Porphyromonas gingivalis, Treponema denticola, and F. nucleatum at the implant sulcus was affected by these microorganisms in the gingival crevice of adjacent teeth rather than those on occluding and contralateral teeth. PMID- 20002683 TI - Detection of candidaemia in patients with and without underlying haematological disease. AB - Diagnosing candidaemia remains difficult despite the development of new diagnostics. We report a direct comparison of three different blood-culture systems and four indirect tests. One hundred and fourteen episodes either with haematological disease and fever despite antibacterials, or with documented invasive candidiasis, were enrolled prospectively. Clinical, para-clinical information and surveillance cultures were obtained. Blood culture was performed using conventional blood-culture bottles, mycosis bottles, and the Isolator 10 lysis centrifugation system. Serum D-arabinitol/L-arabinitol (DA/LA) ratios were determined by gas chromatography mass spectrometry. Antigen, mannan-antigen (Ag) and anti-mannan antibody (Ab) were detected by CandTec, Platelia Candida Ag ELISA and Candida AB/AC/AK kits, respectively. Episodes were classified as proven (n = 24), probable (n = 14), possible (n = 52) or unlikely (n = 24) invasive candidiasis. Candidaemia involved C. albicans (17), C. albicans + C. glabrata (3), C. tropicalis (1) and yeast (1). Mycosis bottles yielded two additional positives and the conventional blood culture yielded one positive not identified by other blood-culture methods. Considering proven and unlikely episodes, respectively, sensitivity and specificity were as follows: mannan-Ag and/or anti mannan Ab: 83.3%, 78.3%; DA/LA ratio: 41.7%, 86.4%; and CandTec Candida Ag: 66.6%, 70.8%. Lowering the cut-off values to mannan-Ag 0.10 ng/mL and anti-mannan Ab 4 AU/mL, the values were: 100%, 73.9%. Applying the DA/LA ratio to only patients with haematological neutropenia the values were: 75%, 90.5%. Fungal blood culture allowed slightly improved detection of candidaemia. The best indirect test performance was obtained from combined mannan-Ag and anti-mannan Ab detection, especially with lower cut-offs. DA/LA ratio appears to be useful in the context of haematological neutropenia. PMID- 20002684 TI - Hard to swallow--emerging and re-emerging issues in foodborne infection. PMID- 20002685 TI - Clostridium difficile in food--innocent bystander or serious threat? AB - Clostridium difficile is a critically important cause of disease in humans, particularly in hospitalized individuals. Three major factors have raised concern about the potential for this pathogen to be a cause of foodborne disease: the increasing recognition of community-associated C. difficile infection, recent studies identifying C. difficile in food animals and food, and similarities in C. difficile isolates from animals, food and humans. It is clear that C. difficile can be commonly found in food animals and food in many regions, and that strains important in human infections, such as ribotype 027/NAP1/toxinotype III and ribotype 078/toxinotype V, are often present. However, it is currently unclear whether ingestion of contaminated food can result in colonization or infection. Many questions remain unanswered regarding the role of C. difficile in community associated diarrhoea: its source when it is a food contaminant, the infective dose, and the association between ingestion of contaminated food and disease. The significant role of this pathogen in human disease and its potential emergence as an important community-associated pathogen indicate that careful evaluation of different sources of exposure, including food, is required, but determination of the potential role of food in C. difficile infection may be difficult. PMID- 20002686 TI - Methicillin-resistant Staphylococcus aureus in food products: cause for concern or case for complacency? AB - The widespread use of antimicrobial agents, in combination with insufficient infection control measures, is the main driver of the current pandemic of antimicrobial resistance in human pathogens. The use of antimicrobials in food animal production also contributes, because resistant organisms and resistance genes can spread from animals to humans by direct contact or through the food chain. An important, traditionally human, pathogen, methicillin-resistant Staphylococcus aureus (MRSA), is currently endemic in many hospitals around the world and has also emerged in the community. Recently, a new reservoir of MRSA has been identified in food production animals and people in contact with these animals. This involves a specific clone, multilocus sequence type 398 (ST398), which has spread extensively among animals. ST398 has also been found in up to 11.9% of retail meat samples in several surveys from different parts of the world, posing a potential threat to human health. PMID- 20002687 TI - Listeriosis: a resurgent foodborne infection. AB - Listeria monocytogenes is the causative agent of human listeriosis, a potentially fatal foodborne infection. Clinical manifestations range from febrile gastroenteritis to more severe invasive forms, including sepsis, meningitis, rhombencephalitis, perinatal infections, and abortions. In recent years, an increasing rate of listeriosis has been reported in several European countries. These increases primarily reflect a higher rate of bacteraemic listeriosis in those > or =65 years of age, and are not otherwise correlated with geography, gender, ethnicity, socioeconomic factors or infectious serotypes. In the late 1980s, an upsurge in listeriosis rates was due to the contamination of a small number of food products. However, a restricted range of strains was responsible for most of the additional cases at that time, and no evidence exists for such a pattern since 2001. From a clinical perspective, the importance of isolating the pathogen as a prerequisite for an accurate epidemiological investigation and ultimately stopping transmission cannot be overemphasized. PMID- 20002688 TI - Much meat, much malady: changing perceptions of the epidemiology of hepatitis E. AB - Hepatitis E, which is caused by hepatitis E virus (HEV), may now be considered a zoonosis as well as an anthroponosis. Pigs, boars and deer have been identified as reservoirs, and their flesh and entrails--as meat and offal--as vehicles of HEV transmission. Shellfish also act as vehicles. Dietary, gastronomic and culinary preferences influence how extensively HEV conveyed by these vehicles can be inactivated before their ingestion by the host. Another route of infection is paved by HEV that is enterically shed by humans and by live animals into the environment. Although anthroponotic transmission of HEV is primarily environmental, zoonotic transmission may proceed along both foodborne and environmental routes. PMID- 20002689 TI - Serious unconventional complications of surgery with stapler for haemorrhoidal prolapse and obstructed defaecation because of rectocoele and rectal intussusception. AB - AIM: Treatment of haemorrhoidal prolapse by stapled haemorrhoidopexy (SH) and obstructed defaecation syndrome with the stapled transanal rectal resection (STARR) technique is becoming increasingly popular with patients and surgeons. Unfortunately, serious complications have been identified. The aim of the present study was to analyse the complications and their treatment to see where they might be avoided and to determine best management. METHOD: All Units of Coloproctology belonging to the Italian Unitary Society of Coloproctology (SIUCP) were asked to return documentation of serious complications following SH and STARR. RESULTS: Forty-six reports were received from 23 centres. Twenty-seven serious complications were reported. Twenty occurred after SH (13 PPH 03, 7 PPH 01) (Endo-surgery Inc., Cincinnati, Ohio, USA) and seven after STARR. Complications were treated by abdominal operation in nine patients [colostomy (3), ileostomy (2), Hartmann's resection (1) and anterior resection (1)]. CONCLUSION: Stapled haemorrhoidopexy and STARR can result in serious complications requiring major surgery for their treatment. PMID- 20002690 TI - The role of cell proliferation and crypt fission in adenoma aggressiveness: a comparison of ileoanal pouch and rectal adenomas in familial adenomatous polyposis. AB - AIM: In patients with familial adenomatous polyposis (FAP), ileoanal pouch cancer is rare whereas rectal cancer is common, despite polyp initiation at the two sites being similar at the molecular level. This study investigated whether the disparity in adenoma aggressiveness reflects underlying differences in histogenesis. METHOD: Normal mucosal biopsies and 2-3 mm adenomas from patients with FAP were dissected into individual crypts. Crypt area, morphology, fission and mitoses were analysed for crypts from pouch, rectum and supra-anastomotic ileum. Immunohistochemistry of similar archival samples was performed for lysozyme, beta-catenin and TP53 expression. RESULTS: The morphology of normal crypts was similar at each site, although crypt area differed. The area of normal pouch crypts was intermediate between rectum and ileum. The area of adenomatous crypts of rectum and pouch was similar, but the latter had increased asymmetrical fission. Crypt mitoses were proportional to area in all tissues, but crypt fission was reduced in adenomatous crypts from the rectum compared with the pouch. Pouch adenomas retained lysozyme expression as seen in normal ileum. Nuclear beta-catenin accumulation was similar, but TP53 expression was increased in rectal adenomas. CONCLUSION: Diminutive polyps from rectum and pouch differ in morphology and proliferation. Aggressiveness in rectal polyps is not conferred by increased crypt proliferation, fission, or activation of the Wnt signalling pathway. Increased TP53 expression suggests other molecular mechanisms may be responsible. While crypt mitoses are proportional to crypt area, the threshold for fission may be site specific, indicating that tissue origin may influence histogenesis and thus malignant potential. PMID- 20002691 TI - Single-incision laparoscopic surgery (SILS) in complex colorectal surgery: a technique offering potential and not just cosmesis. AB - AIM: Single-incision (or port) laparoscopic surgery (SILS) has recently emerged as a method to improve morbidity and cosmetic benefit of conventional laparoscopic surgery. The literature contains two reports of SILS right hemicolectomy, and we report our experience of this technique. METHOD: Seven consecutive, unselected patients underwent SILS retrocaecal appendicectomy, right hemicolectomy, extended right hemicolectomy, colectomy with ileorectal anastomosis, proctocolectomy, anterior resection and restorative proctocolectomy/ileoanal pouch using a single Triport (Olympus Keymed, Southend, UK), conventional instrumentation and nerve block analgesia. Three had undergone previous surgery, two had cancer and two were immunosuppressed. RESULTS: Umbilical, right- and left-iliac fossa SILS was feasible using conventional instruments. Operative time ranged between 23 and 195 min (median 48 min). Four patients tolerated normal diet within 6 h (12-16 h for the remainder). Only one patient required postoperative enteral morphine (10 mg * 4). Discharge occurred between 8 and 90 h (median 16 h) of surgery. A secondary haemorrhage from the ileorectal anastomosis was managed conservatively. CONCLUSION: SILS colorectal resection is feasible and safe when performed by an experienced laparoscopic surgeon and theatre team. It may have advantages over conventional laparoscopic surgery in terms of reduced pain, lower cost, faster recovery and cosmesis. PMID- 20002692 TI - Outcome and long-term function of restorative proctocolectomy for Crohn's disease: comparison to patients with ulcerative colitis. AB - AIM: Restorative proctocolectomy (RPC) is the most common operation for chronic ulcerative colitis (CUC), as it provides excellent functional outcome. However, among patients with Crohn's disease (CD), RPC is generally not recommended, as outcome and long-term function may be poor. Our purpose was to compare matched cohorts of CD and CUC patients to determine whether there are differences in outcome or function. METHOD: We queried our prospectively maintained database of patients who underwent RPC from 1991 to 2008. We identified patients who underwent RPC for CD and compared them with a matched cohort of patients who underwent RPC for CUC. RESULTS: We identified 13 patients with CD (seven women, median age 34 years) and 39 patients with CUC (21 women, median age 35 years). The patients were well matched for gender, clinical and demographic variables. Seven patients (54%) with CD had proctitis, but none had perianal or ileal disease. There were four (30.8%) postoperative complications and no anastomotic leaks. The CD group experienced significantly fewer median daily bowel movements (P = 0.02), incontinence for liquids (P < 0.01) and pouchitis (P < 0.01). With a median follow up of 44 months, pouch excision rate was significantly higher in the Crohn's group (2 vs 0%, P < 0.01). CONCLUSION: In patients with CD, RPC may result in fewer daily bowel movements, less liquid incontinence and a lower incidence of pouchitis compared with CUC patients who undergo RPC. However, risk of pouch loss is higher in patients with CD. Therefore, in properly selected patients with CD, RPC provides an acceptable long-term functional outcome. PMID- 20002693 TI - Long-term functional outcome and quality of life after restorative proctocolectomy with ileo-anal anastomosis for colitis. AB - AIM: The study aimed to evaluate long-term health-related quality of life (HRQOL) and functional outcome in patients who had undergone restorative proctocolectomy with ileo-anal anastomosis (IPAA) for ulcerative colitis and familial adenomatous polyposis. METHOD: A total of 156 patients who underwent IPAA during the period 1984-2003 and who still had an intact pouch were included. The HRQOL score was compared with 4152 individuals from the general Norwegian population using the SF 36 questionnaire, and function was evaluated using the Wexner Continence Grading Scale. RESULTS: One hundred and ten (71%) patients answered the questionnaires, 60 (55%) of whom were men. All except five patients had ulcerative colitis. Median (range) age at interview was 47 (19-66) years, and time after surgery was 12 (2-22) years. The IPAA patients scored slightly, but significantly, lower in four of six SF-36 health domains than the control subjects, adjusted for age and gender. Multiple regression analysis showed frequency of nocturnal defaecation, faecal incontinence and urgency to be independent negative prognostic factors of quality of life. Frequency of defaecation was a median of 7 (3-12) bowel movements during the day and 2 (0-6) at night. The majority had some degree of faecal incontinence, median (range) Wexner score of 8 (0-17), and 40% reported urgency of defaecation necessitating alterations in lifestyle. CONCLUSION: Patients with IPAA reported slightly lower HRQOL rates than the general population and had an inferior functional outcome. PMID- 20002694 TI - Rectourethral fistula after injection sclerotherapy for haemorrhoids, a rare complication. PMID- 20002695 TI - Chemoradiotherapy in the management of primary squamous-cell carcinoma of the rectum. AB - AIM: To assess the efficacy of chemoradiotherapy in the management of primary squamous-cell carcinoma of the rectum. METHOD: Nine patients with primary squamous-cell carcinoma of the rectum were treated with chemoradiotherapy from 1985 to 2007. All patients were female, with a mean age of 54 years (range 41-72 years). The mean distance of the tumour from the anal verge was 6 cm (range 4-10 cm). Seven patients were treated with curative intent (two postoperative, three preoperative, two chemoradiotherapy alone). Clinical stages for the curative group were T1N0M0 (1), T3N0M0 (3), T3N1M0 (1) and T4N0M0 (2). Chemoradiotherapy consisted of pelvic radiation 45-54 Gy in 1.8 Gy/fraction and concurrent 5 fluorouracil and mitomycin C. The mean follow-up duration for the curative group was 39 months (range 15-120 months). RESULTS: All seven patients treated with curative intent achieved local control. A complete pathological response was seen in all patients after preoperative chemoradiotherapy. Two patients did not have surgery and remained free of disease. One patient with gross residual disease after surgery achieved local control. 18-Fluorodeoxyglucose (FDG) avidity was detected in all patients who had FDG-PET scans. Both primary and metastatic tumours demonstrated FDG-avidity. CONCLUSION: Primary squamous-cell carcinoma of the rectum appears to be very sensitive to chemoradiotherapy. The high complete response rate suggests that chemoradiotherapy alone with salvage surgery for local failure should be further explored. FDG-PET scan may have a role to play in the management of this disease. PMID- 20002696 TI - Clinical significance of macroscopic completeness of mesorectal resection in rectal cancer. AB - AIM: Local recurrence after resection of rectal cancer is usually regarded as being due to a 'failure' of surgery. The completeness of resection of the mesorectum has been proposed as an indicator of the 'quality' of the resection. We determined the prognostic value of macroscopic evaluation of rectal cancer resection specimens and the circumferential resection margin (CRM) after curative surgery. METHOD: From 1999 to 2006, the macroscopic quality of the mesorectum and the CRM were prospectively assessed in 127 patients who underwent rectal cancer resection with curative intent (R0+R1). Chemoradiotherapy was administered for 61 tumours staged as locally advanced tumours (T3, T4 and N+). Univariate analysis of time to local recurrence and cancer-free survival were tested (Kaplan-Meier) and multivariate analysis calculated with a Cox regression model. RESULTS: The mesorectum was incomplete in 34 (26.8%) patients. At a median follow up of 34 months (range, 9-96 months), in the group with an adequate mesorectal excision, the cumulative risk of local recurrence at 5 years was 10%. This was 25% if the mesorectum was incomplete (P < 0.01). Five-year cancer-free survival was 65% if the mesorectal excision was adequate and 47% if it was not (P < 0.05). Multivariate analysis identified T status, the CRM and the mesorectal score as independent factors for local recurrence, and T and N status and the mesorectal score as independent factors for disease-free survival. CONCLUSION: The outcome of surgical treatment of rectal cancer is related to the completeness of mesorectal excision. It is a more discriminative prognostic factor than the classic tumour-node-metastasis (TNM) system. PMID- 20002697 TI - Management and outcome of anastomotic leakage after colonic surgery. AB - AIM: Anastomotic leakage remains a key factor for morbidity after colonic surgery. The aim of the study was to analyse the outcome of different therapeutical approaches. METHOD: Of 1731 consecutive patients undergoing colonic resection between 1998 and 2005 at our institution, 67 patients with anastomotic leakage were identified from a prospective database. A logistic regression model was used to determine factors which influenced the therapeutic approach and outcome. RESULTS: The overall anastomotic leakage rate was 3.5%. All patients were re-operated. The anastomosis was resected without restoration of continuity in 31 but preserved in 36 patients. An ileostomy was constructed in 27 of 36 patients with anastomotic leakage after repair or revision of the anastomosis, the remaining nine cases were treated without ileostomy. Five of these latter nine vs three of the 27 patients with ileostomy experienced re-leakage (P = 0.05). The overall mortality was 25%. The Mannheim Peritonitis Index was 17.44 for survivors vs 25.64 for nonsurvivors (P < 0.001). Restoration of intestinal continuity was performed in 95% of the patients with ileostomy and in 88% after Hartmann's procedure. Multivariate analysis identified multi organ failure as the only factor predictive of a fatal outcome (P < 0.001). An ASA-score of more than 2 (P = 0.02) and peritonitis (P = 0.002) were reasons for not preserving the anastomosis. CONCLUSION: Repair or redo of the anastomosis without a protective ileostomy frequently results in failure of the procedure. After Hartmann's operation or split stoma creation a majority of patients undergo restoration of intestinal continuity. PMID- 20002698 TI - CA125 and acute appendicitis. PMID- 20002699 TI - Strict follow-up programme including CT and 18F-FDG-PET after curative surgery for colorectal cancer. AB - AIM: The risk of local recurrence following curative surgery for colorectal cancer (CRC) is up to 50%. A rigorous follow-up program may increase survival. Guidelines on suitable methods for scheduled follow up examinations are needed. This study evaluates a strict follow-up program including carcinogenic embryonic antigen (CEA), chest X-ray, abdominal ultrasound (US), computed tomography (CT) and (18)F-FDG positron emission tomography (FDG-PET). METHOD: A cohort of 132 patients, treated by surgery with curative intent for CRC, was included. Patients were followed prospectively with scheduled controls at 3, 6, 12 and 24 months after curative surgery. CEA, chest X-ray, US, CT and FDG-PET supplemented by clinical examination. The end-point was recurrence. Sensitivity and specificity was estimated 2 years after surgery. RESULTS: Of the 132 patients included in the study, 25 experienced recurrence, detected at scheduled controls (n = 18) and at intervals between them (n = 7). The results of CT and FDG-PET were correlated with recurrence. CT combined with FDG-PET had the highest specificity and sensitivity. CONCLUSION: A total of 72% of recurrences were detected at scheduled controls. The findings supported a strict follow-up program following curative surgery for colorectal cancer. FDG-PET combined with CT should be included in control programs. PMID- 20002700 TI - Laparoscopic Heller myotomy with Dor fundoplication for achalasia: long-term outcomes and effect on chest pain. AB - The aim of the present study was to evaluate the long-term outcomes of laparoscopic Heller myotomy with Dor fundoplication (LHD) and its effect on chest pain. Between June 1995 and August 2009, a total of 35 patients with achalasia underwent an LHD. The symptom scores were calculated by combining the frequency and the severity. Pre- and postoperative evaluations included symptom score, radiology, manometry, and 24-hour pH manometry. Median total symptom score was significantly lower than the preoperative score (19 vs 4, P < 0.001) at a median follow-up of 94 months. Among the 35 patients, 18 (51%) had chest pain. The frequency of chest pain was similar for the pre- and postoperative scores, but the severity tended to be less. Median esophageal diameter (5.4 cm vs 3.5 cm, P < 0.001) and lower esophageal sphincter pressure (41 mmHg vs 8.9 mmHg, P < 0.001) were significantly reduced after surgery. Median age, duration of symptoms, esophageal diameter, and lower esophageal sphincter pressure were similar between patients with and without chest pain prior to surgery. No significant differences were observed between the two groups in terms of amplitude, duration, and frequency of contractions from the findings of postoperative 24-hour esophageal manometry. Chest pain resolved in three patients (17%) and improved in seven patients (39%) after surgery. LHD can durably relieve achalasic symptoms of both dysphagia and regurgitation, and it can be considered the surgical procedure of choice. However, achalasic chest pain does not always seem to be related with patient characteristics and manometric findings. PMID- 20002701 TI - Functional analysis of long-term outcome after Heller's myotomy for achalasia. AB - Treatment of achalasia aims at reducing the pressure of the lower esophageal sphincter (LES) and palliate symptoms. Our objective in this study was to investigate functional changes of the esophagus after Heller myotomy and evaluate their influence on postoperative gastroesophageal reflux and esophageal morphologic changes. Between 1980 and 2003, 216 patients with achalasia underwent Heller myotomy, associated with anterior partial fundoplication (Dor fundoplication). Preoperative and long-term outcome data were collected from these patients at our hospital. The objective was to analyze esophageal functional results after Heller myotomy in the long term. Results were classified as excellent, good, fair, or poor, according to Vantrappen and Hellemans' modified classification. One-year, 2-year, 5-year, 10-year, and 20-year postoperative follow-up information was available in 100% of all patients, 91.7%, 85.1%, 60%, 52.6%, and 45.9%, respectively. There were no perioperative deaths. One year after the surgery, all patients had a significant reduction in symptoms of dysphagia and regurgitation. Five years, 10 years, 15 years, and 20 years after surgery, there were 77.2% of patients (142 in 184), 68.1%, 57.1%, and 54.5%, respectively, who were satisfied (excellent to good) with surgery. No esophageal peristalsis was demonstrated in patients during follow-up. Contractile waves in the body of the esophagus were simultaneous. The difference in the distal esophageal amplitude, the LES relaxation rate, and LES pressures in the anterior wall and/ or two sides was significant (P < 0.05) when compared before and after operation. However, there was no significant difference in the LES length and LES pressure in the posterior side. The change of direction of the LES pressure and the relaxation of LES correlate with long-term outcomes. Postoperative gastroesophageal reflux rates, including nocturnal reflux, increased with time. The percentage of patients whose esophageal diameter became normal or remained mildly increased with time in the first 10 years after surgery changed significantly. Myotomy is an effective way to palliate symptoms in patients with achalasia. Adequate myotomy can lead to reduction of LES pressure in two or three directions, which may facilitate esophageal emptying by gravity. Surgical intervention does not lead to the return of esophageal peristalsis. Functional damage of LES in patients with achalasia is irreversible. PMID- 20002702 TI - Synopsis of investigations into proposed theories on the etiology of achalasia. AB - The current state of research into the etiology of achalasia only allows for speculation. To date, several studies have been performed investigating genetic, immune, and infectious disease mechanisms; however, none of these have been conclusive. Further research into this topic is warranted given the severity of the disease, and it may be possible that all of these mechanisms are involved in the pathophysiology of the disease. PMID- 20002703 TI - CCL17 and CCL22 chemokines within tumor microenvironment are related to infiltration of regulatory T cells in esophageal squamous cell carcinoma. AB - It has been reported that an increased population of regulatory T cells (T-regs) is one of the reasons for impaired anti-tumor immunity. We investigated the frequency of Foxp3(+) T-regs in tumor-infiltrating lymphocytes (TILs) and peripheral blood lymphocytes (PBLs) of patients with esophageal squamous cell carcinoma (ESCC). Furthermore, in order to elucidate the mechanisms behind T-regs accumulation within tumors, we evaluated the relationship between CCL17 or CCL22 expression and the frequency of Foxp3(+) T-regs. CD4(+)CD25(+)Foxp3(+) T-regs as a percentage of CD4(+) cells were counted by flow cytometry. The frequency of CCL17(+) or CCL22(+) cells among CD14(+) cells in tumors was also evaluated by flow cytometry. Moreover, an in vitro migration assay using T-regs derived from ESCC was performed in the presence of CCL17 or CCL22. The frequency of Foxp3(+) T regs in TILs was significantly higher than that in the normal esophageal mucosa (24.6 +/- 10.0 vs 7.1 +/- 5.9%, P < 0.01). The frequency of Foxp3(+) T-regs in PBLs of ESCC patients was significantly higher than that in normal healthy donors (7.0 +/- 4.2 vs 2.5 +/- 1.0%, P < 0.01). Furthermore, the frequency of CCL17(+) or CCL22(+) cells among CD14(+) cells within tumors was significantly higher than that of normal esophageal mucosa, and there was a significant correlation between the frequency of CCL17(+) or CCL22(+) cells and Foxp3(+) T-regs in TILs. In addition, the in vitro migration assay indicated that T-regs were significantly induced to migrate by CCL17 or CCL22. In conclusion, CCL17 and CCL22 within the tumor are related to the increased population of Foxp3(+) T-regs in ESCC. PMID- 20002704 TI - Effect of pantoprazole in patients with chronic laryngitis and pharyngitis related to gastroesophageal reflux disease: clinical, proximal, and distal pH monitoring results. AB - Few studies had evaluated the results of proton pump inhibitors on distal and proximal pH recording using a dual-channel probe. The aim of this study was to determine the clinical and pH-metric effect of treatment with pantoprazole 80 mg for 8 weeks in patients with ear, nose, and throat (ENT) manifestations of gastroesophageal reflux disease associated with pathological proximal acid exposure. We conducted a prospective open study. Patients included had to have chronic pharyngitis or laryngitis, and a pathological gastroesophagopharyngeal reflux. All patients received treatment with pantoprazole 80 mg daily for 8 weeks. One week after the end treatment, patients had a second ENT examination and a 24-hour pH monitoring using dual-channel probe. We included 33 patients (11 men, 22 women). A pathological distal acid reflux was found in 30 patients (91%). After treatment, the improvement of ENT symptoms was found in 51.5% of patients. Normalization of 24-hour proximal esophageal pH monitoring was observed in 22 patients (66%). After treatment, the overall distal acid exposure, the number of distal reflux events, and the number of reflux during more than 5 minutes were significantly decreased (respectively: 19.4% vs 7.2% [P < 0.0001], 62.7 vs 28.4 [P < 0.0001], and 10.4 vs 3.9 [P < 0.0001] ). Similarly, in proximal level, the same parameters were significantly decreased after treatment (respectively: 6.8% vs 1.6% [P < 0.0001], 32.6 vs 8.1 [P < 0.0001], and 3.4 vs 0.6 [P= 0.005] ). Treatment with pantoprazole reduced the frequency and severity of gastroesophagopharyngeal acid reflux in patients with chronic pharyngitis and laryngitis. PMID- 20002705 TI - Determinants of coverage in community-based therapeutic care programmes: towards a joint quantitative and qualitative analysis. AB - One of the most important elements behind the success of Community-based Therapeutic Care (CTC) programmes for the treatment of severe acute malnutrition has been their ability to achieve high levels of coverage. In CTC, coverage is measured using the Centric System Area Sampling (CSAS) method, which provides accurate and reliable estimates of programme coverage as well as information on the primary reasons for non-attendance. Another important feature of CTC programmes is their use of socio-cultural assessments to determine potential barriers to access and to develop context-specific responses. By analysing data on non-attendance provided by CSAS surveys, in conjunction with data from socio cultural assessments, it is possible to identify common factors responsible for failures in programme coverage. This paper focuses on an analysis of data from 12 CTC programmes across five African countries. It pinpoints three common factors (distance to sites, community awareness of the programme, and the way in which rejections are handled at the sites) that, together, account for approximately 75 per cent of non-attendance. PMID- 20002706 TI - Revealing the socioeconomic impact of small disasters in Colombia using the DesInventar database. AB - Small disasters are usually the product of climate variability and climate change. Analysis of them illustrates that they increase difficulties for local development-frequently affecting the livelihoods of poor people and perpetuating their level of poverty and human insecurity-and entail challenges for a country's development. In contrast to extreme events, small disasters are often invisible at the national level and their effects are not considered as relevant from a macroeconomic standpoint. Nevertheless, their accumulated impact causes economic, environmental and social problems. This paper presents the results of an evaluation of the DesInventar database, developed in 1994 by the Network for Social Studies in Disaster Prevention in Latin America. In addition, it proposes a new version of the Local Disaster Index developed in 2005 within the framework of the Disaster Risk and Management Indicators Program for the Americas, with the support of the Inter-American Development Bank. PMID- 20002707 TI - A national system for disseminating information on victims during mass casualty incidents. AB - Immediate provision of information to the public is vital during mass casualty incidents (MCIs). Failure to implement rapidly a communication response system may result in the public overwhelming hospitals. This paper shares Israel's experience in developing and maintaining a national system for supplying information on the location and identification of casualties. ADAM interfaces online with hospitals' patient registration systems, and allows for immediate electronic transfer of designated data. The system permits information centres to access information on which hospital has admitted identified and unidentified casualties. The latter are photographed at the entrance to the hospital and the picture is stored in ADAM. The system enables hospitals and municipalities to ensure immediate availability and accessibility of information and thus (in our belief) mitigate the concerns of family and friends. Use of such an interface system is recommended as an integral element of emergency preparedness. PMID- 20002708 TI - Application of catastrophe loss modelling to promote property insurance in developing countries. AB - Many mega cities in developing countries are exposed to the sources of natural catastrophes, particularly seismic activity. A high level of seismic hazard in some of these places, coupled with a relatively high degree of vulnerability within the built environment, can result in dire human and economic consequences. This paper contains examples of such potentially disruptive factors in relation to Tehran, Iran. It presents preliminary seismic loss estimates for residential buildings in a pilot area of northern Tehran. The paper briefly investigates the effectiveness of risk management measures and loss compensation mechanisms before assessing the feasibility of an insurance-based risk transfer instrument for managing potential seismic losses among residential buildings in Tehran. It goes on to suggest how probabilistic catastrophe loss modelling can help local insurers to manage their portfolios and facilitate risk sharing among insurance companies and households. Finally, the paper addresses the question of how catastrophe loss modelling can help to strengthen the penetration of property insurance in developing countries. PMID- 20002709 TI - Insuring against earthquakes: simulating the cost-effectiveness of disaster preparedness. AB - Ex-ante measures to improve risk preparedness for natural disasters are generally considered to be more effective than ex-post measures. Nevertheless, most resources are allocated after an event in geographical areas that are vulnerable to natural disasters. This paper analyses the cost-effectiveness of ex-ante adaptation measures in the wake of earthquakes and provides an assessment of the future role of private and public agencies in disaster risk management. The study uses a simulation model approach to evaluate consumption losses after earthquakes under different scenarios of intervention. Particular attention is given to the role of activity diversification measures in enhancing disaster preparedness and the contributions of (targeted) microcredit and education programmes for reconstruction following a disaster. Whereas the former measures are far more cost-effective, missing markets and perverse incentives tend to make ex-post measures a preferred option, thus occasioning underinvestment in ex-ante adaptation initiatives. PMID- 20002710 TI - Having a voice and hearing what you want. PMID- 20002711 TI - Troponin assays: more questions than answers? PMID- 20002712 TI - Emergency demand access block and patient safety: a call for national leadership. PMID- 20002713 TI - Review article: the use of pelvic examination within the emergency department in the assessment of early pregnancy bleeding. AB - Early pregnancy bleeding is a common presentation in the emergency setting. Traditionally, its assessment relied on clinical findings, including pelvic examination. However in recent years, ultrasonography and quantitative beta human chorionic gonadotropin assays have gained prominence and are now first-line in diagnosing early pregnancy bleeding. Accordingly, the role of pelvic examination in the acute setting has been increasingly questioned. This article reviews the evidence for the role of pelvic examination in the assessment of early pregnancy bleeding in the ED. A Medline search was conducted and 43 articles were included in this review. Applicable research is largely observational and of a low level of evidence. However, available data indicate that the role of pelvic examination in the assessment of early pregnancy bleeding is limited, providing that there is prompt access to transvaginal ultrasound examination. Pelvic examination does not provide further diagnostic information over ultrasonography used in conjunction with beta human chorionic gonadotropin assays. The routine use of pelvic examination is not supported by the literature. However, when ultrasonography and beta human chorionic gonadotropin testing are unavailable or the results inconclusive, pelvic examination should be considered. Assessment in these instances must focus on identifying possible life-threatening conditions, such as ectopic pregnancy, as well as determining the safety of discharge pending definitive assessment. Speculum examination is indicated in those presenting with severe bleeding or hypotension as removal of obstructing endocervical products can be a crucial resuscitative measure. PMID- 20002714 TI - The 10 mL syringe is useful in generating the recommended standard of 40 mmHg intrathoracic pressure for the Valsalva manoeuvre. AB - OBJECTIVE: The Valsalva manoeuvre (VM) continues to be first-line management for haemodynamically stable supraventricular tachycardia in the acute setting. 40 mmHg of intrathoracic pressure is seen as an essential component of the VM. Anecdotally, blowing into a 10 mL syringe to move the plunger is one method of pressure generation; however, to date its effectiveness has not been tested. The objective of the present study was to assess if blowing into a syringe sufficient to move the plunger could produce the required 40 mmHg of pressure. METHODS: A two-part experimental study tested the pressure required to move the plunger, and sustain that movement for 15 s, in a Terumo syringe. Part one tested a range of syringe sizes. Part two, a repeated measures study, tested the syringe to ascertain if a pressure reduction occurred after repeated use. A sphygmomanometer was attached to the syringe via a 10 cm length of tubing with another length of tubing attached to the sphygmomanometer enabling an investigator to blow into the syringe. RESULTS: In part one, the 10 mL syringe was the only size noted to provide the required 40 mmHg pressure to move the plunger. In part two, the mean for each of the three tests per syringe varied between 37.0 mmHg (95% CI 34.2 39.8) and 40.2 mmHg (95% CI 37.5-43.0). There was no statistically significant fall noted over three uses of the same syringe. CONCLUSION: The present study has demonstrated that blowing into a 10 mL Terumo syringe, to move the plunger, generated 40 mmHg intrathoracic pressure, thereby meeting the recommended intrathoracic pressure for optimum VM performance. PMID- 20002715 TI - Adverse outcomes following emergency department discharge of patients with possible acute coronary syndrome. AB - OBJECTIVE: To determine the proportion of adverse events in patients discharged after ED assessment for possible acute coronary syndrome. METHODS: Prospective observational cohort study enrolling consecutive patients presenting with symptoms suggestive of coronary syndrome. Main outcome was the proportion of adverse coronary events (defined a priori) within 30 days. RESULTS: Of 2627 patients, 1819 (69%) were discharged without a diagnosis of coronary syndrome and 808 (31%) were admitted for further investigation and treatment. Of these, 385 (14.7%) were given a final diagnosis of acute coronary syndrome. On 30 day follow up, 18 of the discharged patients were diagnosed with acute coronary syndrome (0.7%; 95% confidence intervals [CI] 0.4-1.1%), 10 with unstable angina (0.4%; 95% CI 0.2-0.7%) and 8 with non-ST elevation myocardial infarction (0.3%; 95% CI 0.2-0.6%). There were no cases of ST elevation infarction or death. The sensitivity for diagnosis of acute coronary syndromes was 95.5% (95% CI 92.9 97.3%). Average length of stay was 7 h for discharged patients. Forty-six per cent of patients with diabetes and 47% with a past history of coronary disease were discharged. Subsequent outpatient stress testing was performed in 13.6%. CONCLUSIONS: In a large Australian ED, less than 1% of patients presenting with symptoms suggestive of coronary syndrome were discharged and subsequently had a 30 day adverse event. Reducing this proportion by admitting patients with traditional risk factors would markedly increase hospital workload. Opportunities exist to improve both the safety and efficiency of chest pain assessment in the ED. PMID- 20002716 TI - Short-term risk of adverse outcome is significantly higher in patients returning an abnormal troponin result when tested in the emergency department. AB - OBJECTIVE: Describe short-term mortality and hospitalization outcomes in patients undergoing troponin testing in Western Australia tertiary public hospital ED. METHODS: Retrospective cohort study of adults presenting to urban tertiary ED between July 2000 and December 2004, and undergoing troponin inhibitory or tropomyosin binding testing. Patient data relating to ED attendance, troponins, hospitalization and death were linked using probabilistic data linkage. Troponin results were dichotomized (normal/abnormal) according to hospitals' cut-offs across various time periods. Presentations undergoing ED troponin testing, provided that it was not within 28 days of a previous discharge involving ED troponin testing, met study inclusion criteria. Logistical regression analyses quantified risk of subsequent hospitalization and death. RESULTS: A total of 119 646 troponin tests were performed during 83 635 ED presentations involving 54 214 patients. For the 73 965 ED presentations meeting the study criteria, 6779 (9.2%) recorded an abnormal peak troponin level and 168 (2.5%) of these presentations were discharged directly from ED. After adjusting for covariates, those discharged directly from ED with abnormal troponin results were significantly more likely to die within 2 days (OR 60.4, 95% CI 31.9-114.2), 7 days (OR 31.2, 95% CI 17.9-54.3) and 28 days (OR 19.4, 95% CI 12.5-30.1) of discharge, compared with those with normal troponin results. Adjusted odds of subsequent hospital admission within 28 days following discharge from ED were 3.3 (95% CI 2.4-4.5) times greater for presentations with abnormal versus normal troponin results. CONCLUSION: An abnormal ED troponin result appears to be associated with increased risk of subsequent hospitalization and death within 28 days of discharge. PMID- 20002717 TI - Prevalence of access block in Australia 2004-2008. AB - OBJECTIVE: Access block is the inability of ED patients requiring admission to access appropriate inpatient beds in a timely fashion, defined in Australasia as more than 8 h in the ED. The present study describes changes in prevalence of access block in Australia over a 4 year period. METHODS: Email, telephone and fax survey of ED on six Mondays at 10.00 hours (31 May, 30 August 2004, 18 June, 3 September 2007, 2 June, 2 September 2008). Data collected included point data on the status of patients in the ED at the index time and of recent ED attendance numbers. Results were collated and analysed by state and hospital role delineation. RESULTS: Forty-eight (60%) of 80 eligible ED answered all six surveys. Presentations to the ED the day before rose 15% (P < 0.0001, paired t test) in 4 years, and nationally access block patients in the ED rose an average of 27%, and patients waiting to be seen by a doctor 31%. There were differences between states, with hospitals in New South Wales reporting a significant reduction in access block patients (-51%, P= 0.0002), but all other states a significant increase (+45%, P= 0.001). There were differences by role delineation, with non-paediatric major referral hospitals experiencing the greatest access block, but smaller hospitals experiencing the greatest increase in patients waiting. CONCLUSIONS: Around one-third of all patients receiving care in these ED surveys were experiencing access block. There is evidence that flow through New South Wales ED has improved. The data suggest that most hospitals have passed the point of efficiency. PMID- 20002718 TI - Debriefing critical incidents in the paediatric emergency department: current practice and perceived needs in Australia and New Zealand. AB - Anecdotally critical incident debriefing (CID) is an important topic for staff in paediatric ED. The present study aimed to determine current baseline CID practices and perceived needs of ED staff. A questionnaire regarding CID practice was circulated to all 13 Paediatric Research in Emergency Departments International Collaborative (PREDICT) sites in Australia and New Zealand (including all tertiary paediatric ED), and completed by 1 senior doctor and 1 senior nurse. All PREDICT sites participated (13 nurses, 13 doctors). Seventy per cent did not currently have a hospital protocol on debriefing and 90% did not have ED-specific guidelines. The most commonly debriefed topics were death of a patient, multi-trauma and sudden infant death syndrome, also ranked highest in importance for debriefing. The median reported debriefs per department were 4 per year (range 0-12), all conducted within a week of the CI with half within 24 h. ED workers most likely to be invited to the CID session were doctors, nurses and social workers (96%). Debriefing was mostly conducted internally (62%) and most likely facilitated by a doctor (81%) or nurse (54%). Debriefing addressed both clinical and emotional issues (89%) within the same session (69%). Debriefing was rated as very important, median of 8/10 by doctors and 10/10 by nurses. Almost 90% of those surveyed indicated that they would like a CID programme and guidelines for their department. Debriefing is perceived as important by senior ED clinicians, yet few ED have formalized guidelines or programmes. Best-practice guidelines should be developed. PMID- 20002719 TI - Paracetamol versus ibuprofen: a randomized controlled trial of outpatient analgesia efficacy for paediatric acute limb fractures. AB - Paediatric limb fracture is a common injury that presents frequently to the ED. The primary objective of the present study was to determine whether ibuprofen provides better analgesia than paracetamol for paediatric patients discharged with acute limb fractures. A prospective, randomized controlled study was conducted in a children's ED. Children aged 5-14 years with an acute limb fracture were randomized to be prescribed paracetamol 15 mg/kg/dose every 4 h or ibuprofen 10 mg/kg/dose every 8 h. Objective (child-reported) pain scores using the 'Faces' pain scale were measured over a 48 h period. Child-reported pain did not differ significantly between the paracetamol and ibuprofen groups (mean pain score paracetamol 2.8 [95% CI 2.4-3.4] vs 2.7 [95% CI 2.1-3.3], P = 0.73). Parent reported sleep quality did not differ between the two groups (P = 0.78). Child reported pain score decreased over the 48 h of measurement (P < 0.0001). There were no significant differences in side-effects detected between the two groups. The present study shows that in the outpatient paediatric population, ibuprofen does not provide better analgesia than paracetamol. Pain from an acute fracture can be managed by regular simple oral analgesia and immobilization. PMID- 20002720 TI - Consequences of an unrecognized measles exposure in an emergency department. AB - A recent measles case in a paediatric hospital ED resulted in 111 individuals (patients, family members and health-care workers) potentially being exposed to measles. This report documents the efforts taken to contact trace and provide best practice care for all those exposed to the index case. It also provides a snapshot of community prevalence information on immunity to measles. One hundred per cent of contacted children (n = 24) eligible for vaccination were immunized, whereas 96% of adults surveyed or tested (n = 44) had assumed or proven immunity. However, six infants aged between 6 and 9 months were exposed and might have been a sufficiently large reservoir to facilitate the ongoing spread of measles in the community, if contact tracing and preventative measures had not occurred. This scenario also highlights the need to consider measles in the ED, particularly among travellers, with urgent isolation of suspected cases in the ED according to guidelines discussed. PMID- 20002721 TI - Current level of training, experience and perceptions of emergency physicians as expert witnesses: a pilot study. AB - The objective of the present study was to determine emergency physicians' training, experience and perceptions as expert witnesses. Emergency physicians of an adult tertiary referral and teaching hospital participated in a pilot survey regarding their experiences in report writing and in court as expert witnesses. The 28-item survey also examined the amount of formalized forensic medical teaching that emergency physicians had received during their training. Of the participants, 41% (95% CI 21.6-64.1; 7/17) had never received any undergraduate or postgraduate training in forensic medicine, 11/17 (65%, 95% CI 41.2-82.8) had provided a written expert opinion for court, and 12/17 (71%, 95% CI 46.6-87.0) had attended court as an expert witness. All participants considered themselves 'skilled in attending an emergency resuscitation', whereas 3/13 (23%, 95% CI 7.5 50.9) considered themselves 'skilled in attending a courtroom trial'. Nearly 90% (95% CI 64.7-98.0; 15/17) thought that medical evidence training should be a requirement of emergency speciality training. The most commonly preferred forms of medical evidence training were mock court sessions (76%, 95% CI 52.2-91.0; 13/17) and forensic workshops (76%, 95% CI 52.2-91.0; 13/17). From 10 non technical skills required of an EP, 'appearing in court as an expert witness' was perceived to be the second most difficult skill by most respondents. Emergency physicians in this pilot study have limited training for the role of expert witness and see it as one of the most difficult non-technical skills they have to perform. Further research is required regarding the current and future scope of forensic training. PMID- 20002722 TI - Emergency physicians as expert witnesses: 'from frontline wise to courtroom woes'. AB - A 'Medical Expert' witness is one who has been accepted by the court as having special knowledge of the subject about which they are testifying and as a result is allowed to give evidence of their medical opinion. In contrast, ordinary witnesses can only give evidence regarding facts that they have observed. Emergency medicine is one of a few speciality areas of medicine more likely to be involved in legal proceedings. Preparing the emergency physician to be an expert witness is an invaluable part of their training. Education in this area should concentrate on the purpose of the evidence, the legal framework in which it is delivered and the specific skill requirements of medical expert witnesses. PMID- 20002723 TI - Queensland emergency medical system: a structural and organizational model for the emergency medical system in Australia. AB - OBJECTIVE: The emergency medical system (EMS) can be defined as a comprehensive, coordinated and integrated system of care for patients suffering acute illness and injury. The aim of the present paper is to describe the evolution of the Queensland Emergency Medical System (QEMS) and to recommend a strategic national approach to EMS development. METHODS: Following the formation of the Queensland Ambulance Service in 1991, a state EMS committee was formed. This committee led the development and approval of the cross portfolio QEMS policy framework that has resulted in dynamic policy development, system monitoring and evaluation. This framework is led by the Queensland Emergency Medical Services Advisory Committee. RESULTS: There has been considerable progress in the development of all aspects of the EMS in Queensland. These developments have derived from the improved coordination and leadership that QEMS provides and has resulted in widespread satisfaction by both patients and stakeholders. CONCLUSIONS: The strategic approach outlined in the present paper offers a model for EMS arrangements throughout Australia. We propose that the Council of Australian Governments should require each state and Territory to maintain an EMS committee. These state EMS committees should have a broad portfolio of responsibilities. They should provide leadership and direction to the development of the EMS and ensure coordination and quality of outcomes. A national EMS committee with broad representation and broad scope should be established to coordinate the national development of Australia's EMS. PMID- 20002724 TI - Endovascular management of an axillary arterial injury post shoulder dislocation. AB - Axillary arterial injury is a rare complication of non-recurrent glenohumeral joint dislocation. Previously this type of injury would have required open exploration of the axillary artery with associated risk of iatrogenic injury to the axillary vein or brachial plexus. We present the case of a 75-year-old lady, who following a fall in her own home, sustained a dislocation of her right shoulder joint complicated by avulsion of a branch of her axillary artery. We describe the successful endovascular management of the injury using a self expanding stent and propose this as the preferred surgical option where the artery is not completely transected. PMID- 20002725 TI - Clinical challenges and serious vascular complications in neurofibromatosis. AB - Although generally regarded as a benign and chronic condition, neurofibromatosis can be associated with acute vascular and neurological complications. We report the case of a patient with neurofibromatosis who developed massive haemothorax as a result of spontaneous rupture of the left vertebral artery and left subclavian artery. The case was a diagnostic and management challenge. PMID- 20002726 TI - LIPID REGISTRY: intravenous lipid emulsion as antidote. PMID- 20002727 TI - It's not all science. PMID- 20002730 TI - P53 deletion may drive the clinical evolution and treatment response in multiple myeloma. AB - We report a patient with multiple myeloma presenting with a paraspinal plasmacytoma with a marked dissociation between the response obtained in bone marrow (BM) infiltration and that achieved in soft tissue masses. While a complete remission was reached and maintained in BM, extramedullary plasmacytomas were refractory to every line of treatment. Genetic analysis identified the presence of t(4;14) and RB deletion in myeloma cells of both origins. However, a P53 deletion was only detected in plasma cells from extramedullary plasmacytomas. This finding suggests that P53 deletion has a role in the lack of treatment response of extramedullary plasmacytomas. PMID- 20002732 TI - Safe and feasible outpatient treatment following induction and consolidation chemotherapy for patients with acute leukaemia. AB - Traditionally, patients with acute leukaemia are admitted to hospital during chemotherapy-induced pancytopenia, although a few recent reports have reported the feasibility and safety of outpatient treatment. We have developed an outpatient treatment programme for patients with acute leukaemia incorporating comprehensive patient education for self-care management at home during pancytopenia and involvement of patients in care of their tunnelled central venous catheter (CVC). During neutropenia, patients are treated with prophylactic ciprofloxacine, amoxicillin/clavulanic acid and fluconazole. Herein, we report the results of outpatient treatment of 60 patients with acute leukaemia (54 with acute myeloid leukaemia) followed prospectively in the period from March 2004 to 2007. After induction chemotherapy, outpatient treatment was possible after 48 of 73 induction courses, with no readmission in 19 of these (40%). A total of 129 consolidation courses were administered with outpatient treatment following 116 of these, with no readmission in 69 (59%). The median number of days spent at home with neutrophils below 0.5 x 10(9)/L was 8 d per course following induction and 12 d following consolidation chemotherapy. The predominant cause of readmission was neutropenic fever, in most instances of unknown origin. Coagulase negative staphylococci and Enterococcus faecium were the most frequently identified bacteria in blood cultures, whereas only four positive blood cultures with multiresistant Escherichia coli were identified in the entire patient cohort, the latter exclusively observed in patients receiving antibiotic prophylaxis. The majority of the patients were able to take care of their CVC including change in dressing and heparin flushing. There were 12 CVC-related infections. There were no treatment-related deaths. We conclude that outpatient treatment of patients with acute leukaemia is feasible and safe. PMID- 20002731 TI - MYH9 related disease: four novel mutations of the tail domain of myosin-9 correlating with a mild clinical phenotype. AB - MYH9-related disease (MYH9-RD) is a rare autosomal dominant disorder caused by mutations in MYH9, the gene encoding the heavy chain of non-muscle myosin IIA. All patients present congenital macrothrombocytopenia and inclusion bodies in neutrophils. Some of them can also develop sensorineural deafness, presenile cataract, and/or progressive nephropathy leading to end-stage renal failure. We report four families, each with a novel mutation: two missense mutations, in exons 31 and 32, and two out of frame deletions in exon 40. They were associated with no bleeding diathesis, normal, or only slightly reduced platelet count and no extra-hematological manifestations, confirming that alterations of the tail domain cause a mild form of MYH9-RD with no clinically relevant defects. PMID- 20002735 TI - Anticipation of age at onset in familial multiple sclerosis. AB - BACKGROUND AND OBJECTIVE: Anticipation of age at onset in the younger generations is a widely known characteristic of many diseases with genetic inheritance. This study was performed to assess whether there is anticipation of age at onset in younger generations of familial multiple sclerosis (MS) in a Spanish population and to compare clinical characteristics of familial and sporadic MS. METHODS: We studied a cohort of 1110 patients diagnosed with MS and followed-up in our MS Unit. Patients were considered as familial MS if they had in their family at least one relative of first or second degree diagnosed with MS. Otherwise, patients were considered to have sporadic MS. We compared the age at onset between relatives from different generations, and we also compared the age at onset of familial and sporadic MS. RESULTS: A lower age at onset in the younger generations was found (median 22 years vs. 30 years, P < 0.001) and a significant lower age at onset of the disease in familial MS comparing to sporadic MS (median 25 years vs. 29 years, P = 0.042). CONCLUSIONS: There is an anticipation of the age at onset of MS in the younger generations of patients with familial MS. There is also a lower age at onset in familial versus sporadic MS. PMID- 20002733 TI - The GH-IGF system in amyotrophic lateral sclerosis: correlations between pituitary GH secretion capacity, insulin-like growth factors and clinical features. AB - BACKGROUND AND PURPOSE: The growth hormone (GH) and insulin-like growth factor (IGF) system may be involved in neurodegenerative processes, and some abnormalities have been reported in amyotrophic lateral sclerosis (ALS). Our aim was to investigate the GH-IGF axis in patients with ALS and evaluate correlations between this endocrine system and clinical features. METHODS: Serum levels of GH, IGF-I, IGF-II, insulin, IGF-binding protein 1 (IGF-BP1), and IGF-binding protein 3 (IGF-BP3) were measured in 25 patients with ALS and 25 age-, gender-, and BMI matched healthy controls. A GHRH plus arginine test was performed in patients and controls. Clinical status of patients was evaluated with the ALS Functional Rating Scale - Revised (ALSFRS-R) and upper motor neuron (UMN) score. RESULTS: GHRH plus arginine test showed GH deficiency (GHD) in 13 (52%) patients with ALS; severe GHD was found in 6 (24%) and partial GHD in 7 (28%) patients. IGF-I levels were significantly higher in patients with ALS than in healthy controls (182.9 +/ 90.8 vs. 139.4 +/- 58.1 ng/ml; P = 0.015). IGF-I levels were higher in patients with ALS with UMN score >10 than those with UMN score <10 (217.8 +/- 100.8 vs. 155.5 +/- 74.6 ng/ml, P = 0.05). IGF-II levels were significantly lower in patients with ALS than in healthy controls (720.9 +/- 215 vs. 1001.9 +/- 475.4 ng/ml; P = 0.03). CONCLUSIONS: The results demonstrate an impairment of the GH IGFs system in ALS. The degenerative process in ALS might lead to a compensatory increase in IGF-I in an attempt to provide additional support to motor neurons or degenerating muscle fibers. The decrease in IGF-II levels may also be of pathological significance. PMID- 20002738 TI - Current status of the use of Botulinum neurotoxin type A. PMID- 20002739 TI - Routine use of Xeomin in patients previously treated with Botox: long term results. AB - BACKGROUND AND PURPOSE: Based upon large and carefully performed studies Xeomin was first registered in 2005. However, its real potential can only be assessed, when it is used outside of study design restrictions, in an independent setting, in off-label indications and during continued use. METHODS AND RESULTS: Two hundred and sixty-three patients (91 with dystonia, 84 with spasticity, 17 with hemifacial spasm and re-innervation synkinesias, 64 with hyperhidrosis, 7 with hypersalivation), who were previously treated with Botox for at least 1 year under stable conditions, were converted in a blinded fashion to Xeomin using a 1:1 conversion ratio and identical treatment parameters. Therapeutic outcome and adverse effects were monitored by neurological examination and structuralised interviews. In 223 patients (all except those with axillary hyperhidrosis) Xeomin was used continuously throughout a 3 year period. Altogether 1050 injection series were performed. Patients with dystonia received 261.5 +/- 141.0 MU Botox/Xeomin, patients with spasticity 450.5 +/- 177.1 MU, patients with hemifacial spasm and reinnervation synkinesias 44.7 +/- 19.5 MU and patients with hyperhidrosis 286.9 +/- 141.6 MU. The maximum botulinum toxin dose applied was 840 MU. There were no subjective or objective differences between Botox and Xeomin treatments with respect to onset latency, maximum and duration of their therapeutic effects and their adverse effect profiles. Long-term use did not reveal additional safety relevant aspects. None of the patients lost therapeutic efficacy during the observation period. CONCLUSIONS: Xeomin can be used safely in doses of up to 840 MU. Even when applied in high doses it did not produce secondary therapy failure. There were no diffusion differences between Botox and Xeomin. Using a conversion ratio of 1:1 Xeomin and Botox can easily be exchanged in a continued treatment. PMID- 20002740 TI - Xeomin in the treatment of cervical dystonia. AB - BACKGROUND AND PURPOSE: Botulinum toxin type A (BoNT/A) is a highly effective and well-tolerated treatment for focal dystonias. The BoNT/A in Botox and Dysport is part of a high-molecular-weight complex that contains hemagglutinins and other non-toxic proteins, whilst Xeomin is a highly purified BoNT/A free of such complexing proteins. In the largest controlled study of BoNT/A published to date (Neurology 2005; 64: 1949), it was demonstrated that Xeomin is non-inferior to Botox and has 1:1 efficacy in the treatment of cervical dystonia. A possible limitation of continued BoNT/A treatment is antibody development. Based on its physiochemical properties and toxicological evidence, Xeomin is expected to have a reduced incidence of non-responders after long-term treatment compared with other marketed BoNT/A products. METHODS AND RESULTS: In our ongoing open-label study, 100 patients suffering from cervical dystonia are continuously treated with Xeomin; 50 patients were treated de novo, the remaining patients had been previously treated with Botox, Dysport or NeuroBloc/Myobloc. All patients showed negative results in antibody testing at the beginning of Xeomin treatment. During continuous treatment with Xeomin up to 2 years, patients continued to respond well to Xeomin treatment. CONCLUSION: The treatment was well tolerated and no patient has developed neutralizing antibodies as measured using the sensitive mouse hemidiaphragma assay within these first 2 years. PMID- 20002741 TI - Xeomin: an innovative new botulinum toxin type A. AB - Botulinum toxin type A (BoNT/A) is a well-established treatment for conditions characterized by muscle and autonomic nerve terminal overactivity, such as cervical dystonia and blepharospasm, and hyperhidrosis, respectively. BoNT/A is not digested in the gastrointestinal tract as it forms a complex with several proteins that protect and stabilize the neurotoxin. However, the pure neurotoxin is solely responsible for the therapeutic effect, and the complexing proteins have been shown to exhibit immunostimulating activity. The complexing proteins are not required for the stabilization of the neurotoxin in a formulation; the complexing proteins immediately dissociate from the neurotoxin at a physiologic pH, so they do not influence the spread of the neurotoxin. Xeomin is the only botulinum toxin that is free from complexing proteins and is stable at room temperature for a period of 4 years. When injected directly into muscles, Xeomin inhibits local neuromuscular cholinergic transmission, causing focal weakness. It binds to motor nerve terminal pre-synaptic receptors, is internalized via receptor-mediated endocytosis and then selectively cleaves a protein called SNAP 25. This is one of several so-called 'SNARE' proteins involved in exocytosis. Cleavage of SNAP-25 inhibits the secretion of acetylcholine causing the paralysis of the muscle. The clinical effects begin 24-72 h after injection, peak at approximately 4-6 weeks and are sustained for several months. PMID- 20002742 TI - Clinical efficacy and tolerability of Xeomin in the treatment of blepharospasm. AB - BACKGROUND AND PURPOSE: Blepharospasm is classified as a focal dystonia, and botulinum toxin type A (BoNT/A) has been shown to be a highly effective and well tolerated symptomatic treatment. Xeomin, the latest addition to BoNT/A preparations, is a purified, freeze-dried BoNT/A that is free from complexing proteins. METHODS AND RESULTS: In a double-blind, parallel-group, multicentre study, 300 patients with blepharospasm received either Xeomin or Botox 15-80 U (J Neural Transm 2006; 113: 303). Both treatments produced statistically significant improvements from baseline in the Jankovic Rating Scale at week 3 (primary efficacy variable; Xeomin: -2.90; Botox: -2.67; P < 0.0001 from baseline for both), with the difference between treatments (-0.23) indicating that Xeomin was clinically non-inferior to Botox. No significant differences were found between Xeomin and Botox for all secondary variables. There were no clinically relevant differences between Xeomin and Botox in safety parameters, with 40 of 148 patients (27.0%) treated with Xeomin reporting adverse events versus 45 of 155 patients (29.0%) treated with Botox. The most common adverse event was ptosis (6.1% Xeomin and 4.5% Botox). CONCLUSION: Clinical evidence to date suggests that Xeomin is an effective treatment for blepharospasm that does not differ from Botox in terms of its potency, duration of effect or adverse reaction profile. PMID- 20002743 TI - 8th Annual Meeting of the German Dermatoendocrinology Working Group (ADE) of Arbeitsgemeinschaft Dermatologische Forschung (ADF), 4.3.2009, Heidelberg. PMID- 20002745 TI - Structural and serological studies of the O-polysaccharide of strains from a newly created Proteus O78 serogroup prevalent in Polish patients. AB - Seven Proteus mirabilis strains from five Polish patients (five isolates from urea and two from feces) appeared to be a bacterial clone widespread in hospitals, most probably due to nosocomial infection and autoinfection. Enzyme linked immunosorbent assay and Western blot showed that lipopolysaccharides from all strains studied are serologically identical to each other but distinct from Proteus lipopolysaccharides studied earlier and, hence, these strains could not be classified in any of the currently existing 77 Proteus O-serogroups. Accordingly, structural analysis of the O-polysaccharide of a representative strain 1B-m revealed a structure that is typical of Proteus O-antigens but is unique in detail. Based on these data, we propose to classify the strains studied as a new serogroup in the genus Proteus named O78. PMID- 20002746 TI - A comparative study of RNA and DNA as internal gene expression controls early in the developmental cycle of Chlamydia pneumoniae. AB - Many microbial pathogens invade and proliferate within host cells and the molecular mechanism underlying this behavior is currently being revealed for several bacterial species. Testing clinically relevant antibacterial compounds and elucidating their effects on gene expression requires adequate controls, especially when studying genetically intractable organisms such as Chlamydia spp., for which various gene fusions cannot be constructed. Until now, relative mRNA levels in Chlamydia have been measured using different internal gene expression controls, including 16S rRNA, mRNAs, and DNA. Here, we compared the advantages and disadvantages of various internal expression controls during the early phase of Chlamydia pneumoniae development. The relative abundance of target mRNAs varied using the different internal control RNAs. This was partly due to variation in the transcript stability of the RNA species. Also, seven out of nine of the analyzed RNAs increased fivefold or more between 2 and 14 h postinfection, while the amount of DNA and number of cells remained essentially unaltered. Our results suggest that RNA should not be used as a gene expression control during the early phase of Chlamydia development, and that intrinsic bacterial DNA is preferable for that purpose because it is stable, abundant, and its relative amount is generally correlated with bacterial numbers. PMID- 20002748 TI - A novel carbon source-dependent genetic transformation system for the versatile cell factory Hypocrea jecorina (anamorph Trichoderma reesei). AB - Genetic transformation is an indispensable tool for basic fungal research, as well as a useful technique for directed improvement of industrial strains. Here we describe a simple and reproducible transformation system for the filamentous fungus Hypocrea jecorina. The system is based on hxk1 (encoding hexokinase) as selectable marker, a hexokinase-negative strain and D-mannitol, which is used as selective carbon source and osmotic stabilizer. Following transformation with the hxk1 gene, the obtained transformants were able to grow on D-mannitol as sole carbon source. Transformation efficiency achieved using D-mannitol as carbon source and osmotic stabilizer was roughly five times higher than that using D sorbitol. The utility of this system was further demonstrated by transformation of H. jecorina with the egfp (encoding the enhanced green fluorescent protein) gene. Fluorescence microscopy revealed EGFP fluorescence in positive transformants. Our results demonstrated the feasibility of exploiting a carbon source metabolic pathway for the development of promising fungal transformation systems, which provides a new molecular toolbox for genetic modifications of the cell factory H. jecorina. PMID- 20002749 TI - On the expanding, then contracting scope of scientific publications. PMID- 20002750 TI - Proconvulsant potential of cyproheptadine in experimental animal models. AB - In epileptic patients cyproheptadine is frequently prescribed as an appetite stimulant for the treatment of anorexia associated with anti-epileptic drugs and for the management of 'serotonin syndrome' in depressed epileptic patients. However, the study of serotonergic and histaminergic pathway shows that the decreased neurotransmission of serotonin and histamine in the brain reduces seizures threshold. Since, cyproheptadine interferes with these pathways via antagonizing subtypes of 5-HT(1/2) receptors and H(1) receptor, therefore the present study was undertaken to investigate its effect on seizures threshold, so as to substantiate its use in epileptics. In the present study convulsions were induced in mice by, maximum electroshock (MES), picrotoxin, and pentylenetetrazol (PTZ). Cyproheptadine (4 mg/kg, i.p.) was administered per se and along with clinically used anti-epileptic drugs (phenytoin 25 mg/kg, i.p. and diazepam 5 mg/kg, i.p.) in different groups of mice, onset and extent of convulsions in these groups were compared with that of vehicle control and anti-epileptics per se treated groups. Percentage mortality in all groups was also determined. Results depicted a significant increase in duration of tonic hind limb extension in MES and decrease in latency to clonic convulsions induced by PTZ and picrotoxin in cyproheptadine treated groups (per se and along with anti epileptics), as compared to vehicle control and anti-epileptics per se treated groups respectively. Percentage mortality was also increased with cyproheptadine treatment. Therefore it is concluded that cyproheptadine pretreatment reduces threshold, increases severity of seizures and decreases the efficacy of clinically used anti-epileptic drugs in experimental animal models of convulsions. PMID- 20002751 TI - 'Multiple action fixed combination. Present or future?'. AB - Cardiovascular disease (CVD) is the most common cause of death in Western countries and will continue to be so in upcoming years. A close correlation has been demonstrated among CVD, stroke, ischemic heart disease, renal failure and a number of modifiable risk factors. As cardiovascular (CV) risk factors commonly co-exist, high-risk patients with hypertension, obesity and diabetes may well benefit from a multiple action combination of CV agents with synergistic efficacy. Control of blood pressure (BP) and the other CV risk factors is still far from the optimal rates and achievement of internationally accepted goals must be imperative. The benefits of achieving these goals, including significant reductions in CV morbidity and mortality, are well documented. Thus, a rigorous effort to improve BP goal attainment is required. Most of the patients will need two or more antihypertensives to achieve BP goal. Administering of two drugs in a single-dose formulation substantially improves patient compliance compared with separate agent administration. Fixed-dose combination therapy can offer potential advantages over individual agents, including increased efficacy, reduced incidence of adverse effects, lower healthcare costs and improved patient compliance through the use of a single medication administered once daily. Currently available fixed-dose agents include several combinations with complementary pharmacodynamic activity. Last, the polypill includes several CV acting agents that affects various CV risk factors and offers encouraging results, although more data are needed to strengthen the polypill concept, its efficacy and safety. PMID- 20002752 TI - Comprehensive geriatric assessment of elderly highlanders in Qinghai, China I: activities of daily living, quality of life and metabolic syndrome. AB - AIM: To reveal the comparison of comprehensive geriatric functions of elderly highlanders in Qinghai Plateau in China among three different ethnic groups. METHODS: Activities of daily living (ADL), screening-based depression, quality of life (QOL) and checking-up of metabolic syndrome including community-based oral glucose tolerance test were assessed in 393 community-dwelling elderly subjects aged 60 years or more (247 Han elderly subjects, 49 Mongolian ones and 97 Tibetan ones). RESULTS: Tibetan elderly highlanders were more disabled in ADL, but had higher QOL than Han elderly ones in Qinghai Plateau. Blood pressure measurements, rate of hypertension and hemoglobin concentrations in Tibetan elderly highlanders were lower than Han ones. Rates of diabetes and impaired glucose tolerance in elderly highlanders were relatively lower than other Asian elderly lowlanders. CONCLUSION: Prevalence of metabolic syndrome in elderly highlanders in Qinghai was still not high, however, we should pay attention to its tendency related with socialglobalism in the near future. Further investigation on physiological adaptability to hypoxic environment and human ageing phenomena in a global context may open a new research frontier for ageing science. PMID- 20002753 TI - Comprehensive geriatric assessment of elderly highlanders in Qinghai, China II: the association of polycythemia with lifestyle-related diseases among the three ethnicities. AB - AIM: The objective of this study is to disclose the association of polycythemia with lifestyle-related diseases (hypertension, obesity and glucose intolerance) among the three ethnicities in Qinghai, China. METHODS: The subjects were 393 elderly people (247 Han, 97 Tibetan and 49 Mongolian) aged 60 years and more living in Qinghai (3000 m a.s.l.) in China. The associated factors with polycythemia were analyzed in the subjects. Excessive polycythemia was defined as hemoglobin concentration over 20 mg/dL. RESULTS: Polycythemia was associated with men, hypoxemia, obesity and high diastolic blood pressure (DBP) in the elderly in Qinghai. Male sex was associated with polycythemia in all ethnicities. Obesity was associated with Han and Tibetan men. Glucose intolerance and activities of daily living were not directly associated with polycythemia after adjustment for sex. There were 7.9% with excessive polycythemia. Independently-associated factors for excessive polycythemia were male sex, body mass index of 25 or more, SpO(2) of less than 85%, DBP of 85 mmHg or more and Han ethnicity (vs Tibetan) by multiple logistic regression. CONCLUSION: There was a close association of polycythemia with diastolic hypertension and obesity in lifestyle-related diseases in high-altitude elderly people. Han people had a higher hemoglobin concentration after adjustment of lifestyle-related diseases compared with Tibetan people. The difference of hemoglobin concentration may be due to Tibetans undergoing a much longer period of adaptation than Han people. Further study is needed to disclose the association between the difference of hypoxic adaptation, lifestyle-related diseases and chronic mountain sickness for their prevention. PMID- 20002754 TI - Comprehensive geriatric assessment of elderly highlanders in Qinghai, China, III: oxidative stress and aging in Tibetan and Han elderly highlanders. AB - BACKGROUND: Although there are several factors which may contribute to oxidative stress at high altitude, little is known about the association between oxidative stress and aging in the community-dwelling elderly in the Tibetan Plateau. METHODS: Reactive oxygen species (ROS) and comprehensive geriatric functions were examined among 235 community-dwelling elderly subjects aged 60 years or more (146 Hans and 89 Tibetans). As a marker of ROS, the levels of reactive oxygen metabolites (ROM) were measured using the d-ROM test. RESULTS: The rate of dependence of basic activities of daily living (basic ADL) among Tibetan elderly highlanders was significantly higher than that among Han elderly highlanders. The d-ROM level was higher among the Tibetan elderly than those among the Han elderly (Tibetan 465.6 +/- 97.9 Carr U, Han 415.3 +/- 72.0 Carr U, P = 0.003). The ROM level was higher among women than those among men. Stepwise multiple regression analysis showed that being Tibetan, female, and oxygen saturation were independent predictors of increasing d-ROM level (Tibetan beta, 0.241; female beta, 0.206; oxygen saturation beta, 0.218). The high levels of ROM (d-ROM >500 Carr U) were significantly associated with dependence of basic ADL after adjustment for age, sex and ethnicity (odds ratio = 2.51, P = 0.028). CONCLUSION: The findings of this study imply the possibility that ROS is higher among Tibetan elderly highlanders than that of Han, which related to the geriatric items. Further studies are needed to show the impact of oxidative stress on the aging of highlanders. PMID- 20002755 TI - Comprehensive geriatric assessment of elderly highlanders in Qinghai, China IV: comparison of food diversity and its relation to health of Han and Tibetan elderly. AB - AIM: To examine the association between food diversity and health status of Han and Tibetan elderly highlanders in Qinghai Plateau, China. METHODS: The study population consisted of 240 community-dwelling elderly subjects aged 60 years or more (176 Han elderly subjects, 64 Tibetan ones). Food diversity was determined using an 11-item Food Diversity Score Kyoto (FDSK-11). Subjects were interviewed on health status including activities of daily living (ADL), screening-based depression and quality of life (QOL). Blood chemical investigation was carried out in association with food diversity. RESULTS: ADL was significantly lower in both Han and Tibetan elderly with lower food diversity than those with higher diversity. In Han elderly with lower food diversity, QOL was significantly lower in the items of subjective sense of health, relationship with family and subjective happiness, but not significant in Tibetan elderly. A close association was found between lower food diversity and lower financial satisfaction in both Han and Tibetan subjects. No association was found between food diversity and age or body mass index. Higher food diversity was associated with lower blood glucose level in Han elderly subjects, but the opposite association was found in Tibetan ones. CONCLUSION: Food diversity was associated with ADL and QOL in highlanders in Qinghai, China. Food assessment is very important as a useful indicator to establish the actual condition of diet and its relation to health status of community-dwelling elderly as well as the change of economic background in the Qinghai highlands. PMID- 20002756 TI - Associations between functional tooth number and physical complaints of community residing adults in a 15-year cohort study. AB - BACKGROUND: Oral function influences various general health and organ diseases. We wondered if physical complaints of unknown origin were related to oral function. METHODS: Five thousand five hundred and eighty-four community residents (2206 men and 3378 women, aged 40-89 years) on the Miyako Islands, Okinawa Prefecture, Japan, were recruited for the baseline study in 1987 and followed up for 15 years. Physical complaints and functional tooth numbers were assessed for the baseline study in 1987 by dentists and followed up for 15 years. Physical complaints of unknown origin included shoulder stiffness, lower back pain, pain of upper extremity, pain of lower extremity, neuralgia, ear ringing, difficult of hearing, dizziness and sleeping difficulties. RESULTS: Physical complaints were significantly associated with functional tooth number in the baseline study. There were no systematic differences between physical complaints and functional tooth number among different age groups. Physical complaints did not influence the survival rates in either men or women. CONCLUSION: Oral function may be related to physical complaints. Dental care may be one of the targets to treat these physical complaints of unknown origin. PMID- 20002757 TI - Effects of a robotic walking exercise on walking performance in community dwelling elderly adults. AB - BACKGROUND: Reduced gait speed and stride length are characteristic of gait in elderly people and increase their dependency on assistance. We developed a robotic stride assistance system (SAS) to automatically control the walk ratio during walking. Our aim was to quantify the effects of a walking exercise with the SAS on walking performance and glucose metabolism during walking in community dwelling elderly adults. METHODS: For 3 months, 15 women (72-85 years old) performed the walking exercise twice weekly for 90 min/session. We assessed walking for 5 m at a comfortable speed before and after the intervention. Positron emission tomography with [(18)F]fluorodeoxyglucose (FDG) was used to assess muscle activity during an unassisted 50-min walk. RESULTS: Walking speed was improved by the intervention and FDG uptake by the gluteus minimus, gluteus medius and rectus femoris, and pelvic muscles (iliacus and gluteus muscles) were reduced. CONCLUSION: These results suggest that a walking intervention program using an applied robotic system is useful for improving the walking ability and the efficiency of muscle activities during walking in the elderly. PMID- 20002758 TI - Associated conditions and clinical significance of awake bruxism. AB - BACKGROUND: Awake bruxism is defined as an oral parafunctional activity that includes clenching and grinding of teeth during wakefulness. Confirming the possible related anatomy and the clinical significance of awake bruxism in geriatric hospitals is the aim of this study. METHODS: We analyzed the medical records of 503 patients who were admitted to hospital from April to June 2008. After the recognition of bruxism, the clinical, brain imaging studies and statistical parametric mapping (SPM) of brain single photon emission computed tomography were performed. RESULTS: In each disease group, five of 125 Alzheimer's disease (AD) patients (4.0%), three of 11 frontotemporal dementia (FTD) patients (27.3%), seven of 230 stroke patients (including two patients related to citalopram, 3.0%), one of 45 Parkinson's disease patients (2.2%) and four of 17 hydrocephalus patients (23.5%) had bruxism. Even though awake bruxism occurred early after stroke onset, it occurred late after AD and FTD onset. This occurred in a far advanced stage of AD, while it occurred in a moderately advanced stage of FTD. SPM analysis in AD and FTD patients with awake bruxism revealed significant hypoperfusion in frontotemporal and other subcortical structures. Surface electromyography recordings from the masseter muscle showed rhythmic regular motor activity at a rate of 1-2/s. CONCLUSION: This study suggests that awake bruxism is encountered not infrequently in various diseases in geriatric hospitals. It is frequently observed in FTD and normal pressure hydrocephalus, which characteristically shows frontal lobe dysfunction. These facts and SPM analysis show that awake bruxism can be regarded as a frontal neurological sign of various neurological disorders. PMID- 20002759 TI - Five cases of severe hyponatremia in the elderly successfully treated with low doses of hydrocortisone. AB - Hyponatremia is commonly defined as a serum sodium concentration of 135 mEq/L or less, which is reported to occur in 0.97-2.48% of hospitalized patients. Especially, in elderly patients, severe hyponatremia is sporadically developed without any complaints. Although a variety of causes of hyponatremia are documented, diagnosis of functional adrenal insufficiency is difficult because of a lack of the diagnostic criteria. In this case report, we present five cases of hyponatremia in the elderly, who were presumed to have functional adrenal insufficiency, successfully treated with low doses of hydrocortisone. PMID- 20002760 TI - Peripheral T-cell lymphoma presenting as myelofibrosis with the expression of basic fibroblast growth factor. AB - Myelofibrosis is often observed in chronic myeloproliferative disorders (CMPD), but non-Hodgkin's lymphoma with diffuse myelofibrosis is rare. We describe an elderly case with peripheral T-cell lymphoma-unspecified (PTCL) presenting as diffuse myelofibrosis. Bone marrow biopsy revealed infiltration of atypical lymphocytes and diffuse myelofibrosis without any increase in megakaryocytes. To discuss the pathogenesis of fibrosis, we examined cytokines relative to fibrosis using immunostaining. The expression of basic fibroblast growth factor (bFGF) was diffusely detected in the area of extracellular matrix of bone marrow. In addition, in situ hybridization revealed that infiltrating lymphoma cells expressed bFGF mRNA. Basic FGF, originally identified based on its mitogenicity for fibroblasts, has multiple potential, influencing neoangiogenesis, bone marrow fibrosis and the proliferation of tumor cells. Basic FGF might play an important role in the pathogenesis of myelofibrosis in the present case. PMID- 20002761 TI - Foot care using green tea paste for behavioral and psychological symptoms in dementia patients. PMID- 20002762 TI - Prescribing cascade in an 80-year-old Japanese immigrant. PMID- 20002763 TI - Utilization and expenditures on chiropractic care in the United States from 1997 to 2006. AB - OBJECTIVE: To investigate national utilization and expenditures on chiropractic care between 1997 and 2006. DATA SOURCE: The nationally representative Medical Expenditure Panel Survey (MEPS). STUDY DESIGN: We performed descriptive analyses and generated national estimates from data obtained from U.S. adult (>or=18 years) MEPS respondents who reported having visited a chiropractor (annual sample size between 789 and 1,082). For each year, we examined the estimated total national expenditure, the total number of U.S. adults who received chiropractic care, the total number of ambulatory visits to U.S. chiropractors, and the inflation-adjusted charges and expenditures per U.S. adult chiropractic patient. PRINCIPAL FINDINGS: The total number of U.S. adults who visited a chiropractor increased 57 percent from 7.7 million in 2000 to 12.1 million in 2003. From 1997 to 2006, the inflation-adjusted national expenditures on chiropractic care increased 56 percent from U.S.$3.8 billion to U.S.$5.9 billion. Inflation adjusted total mean expenditures per patient and expenditures per office visit remained unchanged. CONCLUSION: The large increase in U.S. adult expenditures on chiropractic care between 1997 and 2006 was due to a 57 percent increase in the total number of U.S. adult chiropractic patients that occurred from 2000 to 2003. From 2003 to 2006, the total number of U.S. adult chiropractic patients has remained stable. PMID- 20002764 TI - Acute myocardial infarction and coronary artery bypass grafting outcomes in specialty and general hospitals: analysis of state inpatient data. AB - OBJECTIVE: Compare characteristics and outcomes of patients hospitalized in specialty cardiac and general hospitals for acute myocardial infarction (AMI) and coronary artery bypass grafting (CABG). DATA: 2000-2005 all-payor administrative data from Arizona, California, Texas, and Wisconsin. STUDY DESIGN: We identified patients admitted to specialty and competing general hospitals with AMI or CABG and compared patient demographics, comorbidity, and risk-standardized mortality in specialty and general hospitals. PRINCIPAL FINDINGS: Specialty hospitals admitted a lower proportion of women and blacks and treated patients with less comorbid illness than general hospitals. Unadjusted in-hospital AMI mortality for Medicare enrollees in specialty and general hospitals was 6.1 and 10.1 percent (p<.0001) and for non-Medicare enrollees was 2.8 and 4.0 percent (p<.04). Unadjusted in-hospital CABG mortality for Medicare enrollees in specialty and general hospitals was 3.2 and 4.7 percent (p<.01) and for non-Medicare enrollees was 1.1 and 1.8 percent (p=.02). After adjusting for patient characteristics and hospital volume, risk-standardized in-hospital mortality for all AMI patients was 2.7 percent for specialty hospitals and 4.1 percent for general hospitals (p<.001) and for CABG was 1.5 percent for specialty hospitals and 2.0 percent for general hospitals (p=.07). CONCLUSIONS: In-hospital mortality in specialty hospitals was lower than in general hospitals for AMI but similar for CABG. Our results suggest that specialty hospitals may offer significantly better outcomes for AMI but not CABG. PMID- 20002765 TI - Impact of Medicare Part D on Medicare-Medicaid dual-eligible beneficiaries' prescription utilization and expenditures. AB - OBJECTIVE: To examine the effect of Part D on 65-78-year-old noninstitutionalized dual eligibles' prescription utilization and expenditures. DATA SOURCE: Random sample of unique pharmacy customers of a national retail pharmacy chain who filled at least one prescription during both 2005 and 2006. For each subject, we obtained claims data for every prescription filled between January 1, 2005, and April 31, 2007. STUDY DESIGN: Generalized estimating equations were used to examine the experience of a "treatment" group (dual eligibles between 65 and 78 years on January 1, 2005) with that of a "control" group (near-elderly patients with Medicaid coverage between 60 and 63 years on January 1, 2005) during the first 18 months after Part D implementation. PRINCIPAL FINDINGS: Expenditures for the treatment and control groups tracked each other closely in the pre-Part D period. Immediately following the implementation of Part D, expenditures for both groups decreased and then leveled off. There were no significant changes in trends in the dual eligibles' out-of-pocket expenditures, total monthly expenditures, pill-days, or total number of prescriptions due to Part D. CONCLUSIONS: We find no evidence that Part D adversely affected pharmaceutical utilization or out-of-pocket expenditures of dual eligibles during the transition period, nor during the 16 months subsequent to Part D implementation. PMID- 20002766 TI - Clinicopathological characteristics of primary gastric T-cell lymphoma. AB - AIMS: To investigate the clinicopathological characteristics of 20 primary gastric T-cell lymphoma (GTCL) cases without human T-lymphotropic virus type I infection in Japan, a non-endemic area for coeliac disease. METHODS AND RESULTS: Fifteen cases had no history of persistent diarrhoea or severe hypoproteinaemia. Histologically, 13 cases (65%) consisted of large cell lymphoma and seven (35%) were of medium-sized cells. Intraepithelial lymphoma cell invasion was found in three cases (15%). Two of 10 surgical cases (20%) showed intramucosal tumour cell spreading with enteropathy-like features. Helicobacter pylori CagA gene was detected in three of 10 cases (30%). The lymphoma cells of all 20 cases were positive for CD3 and/or TCRbetaF1 and negative for CD56. CD4- and CD8- lymphoma was found in 11 cases (55%), CD4+ lymphoma in seven (35%) and CD8+ lymphoma in two (10%). CD30+, CD5+ and CD25+ lymphomas were detected in nine (45%), 10 (50%) and 11 (55%) cases, respectively. Five-year survival of the 16 available cases was 54%. Early clinical stage and medium-sized cell lymphoma were significantly (P < 0.05) better prognostic factors. CONCLUSIONS: Patients with GTCL exhibit distinct clinicopathological findings and prognoses from those with enteropathy associated T-cell lymphomas. GTCL may be mainly derived from lamina propria and parafollicular T cells. PMID- 20002767 TI - Intravascular lymphocytosis in acute appendicitis: potential mimicry of chronic lymphocytic leukaemia. AB - AIMS: Intravascular lymphocytosis (IVL) in appendectomy specimens is a fairly common phenomenon but has not been described and studied in detail. The aim was to describe the IVL phenomenon in appendiceal specimens removed for appendicitis that can mimic chronic lymphocytic leukaemia and to investigate factors that could contribute to the development of IVL. METHODS AND RESULTS: The clinicopathology of 100 appendectomy cases and the appendices from 20 right hemicolectomy cases were reviewed. The IVL phenomenon was more commonly seen in laparoscopic appendectomy specimens [Fisher's exact 0.011, odds ratio (OR) 3.14] and in patients <38 years old (Fisher's exact 0.012, OR 3.01). CONCLUSIONS: Surgical manipulation and the patient's innate immunity are contributary factors to the development of the IVL phenomenon. Pathologists should be aware of this histological artefact in order to prevent overdiagnosis of intravascular lymphoid neoplasia. PMID- 20002768 TI - Oncocytic versus mitochondrion-rich follicular thyroid tumours: should we make a difference? AB - AIMS: To separate true oncocytic neoplasms from mitochondrion-rich non-oncocytic lesions based on the intracellular relationship between major cell organelles, and to establish the diagnostic and clinical relevance of this distinction. METHODS AND RESULTS: Tissue samples from 276 follicular adenomas, 194 follicular carcinomas, 162 normal thyroids and 296 non-neoplastic lesions were classified as conventional, mitochondrion-rich or oncocytic based on the immunohistochemically assessed quantity and intracellular distribution of mitochondria and endoplasmic reticulum (ER) and nuclear position. Pathological and clinical features were compared among the groups. In oncocytes, densely packed mitochondria resulted in homogeneous immunolabelling of basal cytoplasmic regions, whereas ER and the nuclei were typically displaced to the apical position. This aberrant organelle distribution was not observed in non-oncocytes, which allowed reliable distinction between oncocytic and mitochondrion-rich lesions. Clinically, mitochondrial increase in non-oncocytic lesions was associated with neoplasia, malignancy and higher cancer recurrence rates. Similar correlation, albeit less pronounced, was observed within the oncocytic tumour group. By contrast, oncocytic change per se was not associated with neoplasia, malignancy or cancer aggressiveness. CONCLUSIONS: True oncocytic neoplasms can be distinguished from mitochondrion-rich non-oncocytic tumours based on aberrant distribution of all major cell organelles. This distinction has immediate clinical relevance and should be implemented in practice. PMID- 20002769 TI - Identifying CD82 (KAI1) as a marker for human chromophobe renal cell carcinoma. AB - AIMS: The variation in cytoplasmic staining and growth pattern makes differential diagnosis of chromophobe renal cell carcinoma (RCC), especially from renal oncocytomas (RO), uncertain. The aim was to find an immunohistochemical marker to improve the diagnosis of chromophobe RCC. METHODS AND RESULTS: The expression profile of renal cell tumours was established using the Affymetrix HG U133 Plus 2.0 microarray. After performing hierarchical clustering we selected the best 50 genes and analysed 24 of them by reverse transcriptase-polymerase chain reaction. TMC5, RBM35, SPINK7, CD82/KAI1 and GLIS3 were expressed specifically in chromophobe RCCs. Immunohistochemical studies confirmed the specificity of CD82/KAI1. Analysis of tissue microarrays containing all major types of renal neoplasm revealed immunopositivity for CD82/KAI1 in 78% (69/88) of chromophobe RCCs but in only 3% (6/220) of conventional or clear cell RCCs. All 90 ROs and 171 papillary RCCs were immunonegative for CD82/KAI1. To confirm the specificity of antibody by immunoblotting, we transfected HEK293 cells with an expression vector containing the full-length CD82/KAI1. CONCLUSIONS: CD82/KAI1 is an excellent marker for distinguishing chromophobe RCCs from other types of renal cell tumours, especially from ROs with overlapping phenotype. PMID- 20002770 TI - Overexpression of RAD51 occurs in aggressive prostatic cancer. AB - AIMS: To test the hypothesis that, in a matched series of prostatic cancers, either with or without BRCA1 or BRCA2 mutations, RAD51 protein expression is enhanced in association with BRCA mutation genotypes. METHODS AND RESULTS: RAD51 expression identified immunohistochemically was compared between prostatic cancers occurring in BRCA1 or BRCA2 mutation carriers and controls. RAD51 protein expression in the cytoplasm and nuclei of the benign tissues was significantly less than in the malignant tissues (P < 0.001). In all cancers, cytoplasmic expression of RAD51 was more prevalent and associated with higher Gleason score (P < 0.05) irrespective of BRCA mutational status, than its expression in benign tissues (P < 0.001). Although nuclear immunoreactivity was not observed in BRCA associated cancers with Gleason score < or =7, it was significantly increased in all other groups of prostatic cancers when compared with benign tissues (P < 0.001). CONCLUSIONS: RAD51 protein is strongly expressed in high-grade prostatic cancers, whether sporadic or associated with BRCA germ-line mutations. Distinct localization of RAD51 between cytoplasm and nucleus, particularly in cancers of Gleason score < or =7, reflects distinct levels of RAD51 regulatory activity, from transcription to DNA repair. This biomarker may be of value in identifying patients requiring urgent treatment at diagnosis as well as in analysing biological mechanisms underlying aggressive phenotype of human prostatic cancer. PMID- 20002771 TI - Validation of potential therapeutic targets in alveolar soft part sarcoma: an immunohistochemical study utilizing tissue microarray. AB - AIMS: The molecular signature of alveolar soft part sarcoma (ASPS) is a specific der(17)t(X;17)(p11.2;q25) translocation, resulting in a chimeric transcription factor (ASPSCR1-TFE3). When this disease is no longer amenable to surgical curative intervention, uniformly efficacious therapies are lacking. The aim of this study was to evaluate the expression of potential molecular therapeutic targets in a cohort of ASPS tumour samples. METHODS AND RESULTS: Immunohistochemical analysis for hepatocyte growth factor, c-Met, phosphorylated c-Met, phosphorylated AKT, phosphorylated MEK, epidermal growth factor receptor (EGFR), vascular endothelial growth factor (VEGF), p53 and vimentin was performed on an ASPS tissue microarray, yielding complete data from 26 tumours. Activation of c-Met and its downstream effectors was noted, whereas only limited EGFR expression was seen. VEGF was expressed to varying degrees. Only one sample exhibited strong nuclear p53 expression, while 10 expressed low levels. Vimentin expression was negative in the vast majority of samples (96%). CONCLUSIONS: There is a crucial need for better anti-ASPS therapies. Activated c-Met and the phosphorylation of its downstream effectors validate an intact signalling cascade probably induced by the ASPSCR1-TFE3 chimeric transcription factor. The angiogenic phenotype of these tumours is supported by increased angiogenic factor expression. Combination therapies targeting both tumour cells and angiogenesis merit further investigation. PMID- 20002772 TI - The accuracy of frozen section diagnosis in apparent early ovarian cancer- results from a UK centre. PMID- 20002773 TI - Granulomatous variant of allergic proctocolitis. PMID- 20002774 TI - Digitalization of post-mortem coronary angiography. PMID- 20002775 TI - The proteins ARVCF and p0071 in renal cell carcinomas and their potential use in the diagnosis of renal tumours. PMID- 20002776 TI - Determinants of late HIV diagnosis among different transmission groups in Finland from 1985 to 2005. AB - OBJECTIVES: To study determinants of late HIV diagnosis in a low-HIV-prevalence (<0.1%) country where HIV spread among men who have sex with men (MSM) and heterosexuals in the 1980s, and among injecting drug users (IDUs) in the late 1990s. METHODS: Newly diagnosed HIV cases referred to the Helsinki University Central Hospital between 1985 and 2005 were reviewed to identify determinants of late HIV diagnosis, defined as diagnosis when the first CD4 count was <200 cells/microL, or when AIDS occurred within 3 months of HIV diagnosis. Determinants of late diagnosis were analysed using multivariate logistic regression. RESULTS: Among 934 HIV cases, 211 (23%) were diagnosed late. In the first 4-year interval of each sub-epidemic (1985-1989 for MSM and heterosexuals, 1998-2001 for IDUs), rates of late HIV diagnosis were 13%, 18% and 6%, respectively, but increased thereafter to 29%, 27% and 37%. Late diagnosis was associated with non-Finnish ethnicity, older age, male gender, lack of earlier HIV testing, diagnosis at health care settings and later stage of the sub epidemic. CONCLUSIONS: The lower rate of late diagnosis in the first 4-year interval of each HIV sub-epidemic suggests that the early stages of the HIV epidemic in Finland were detected early. This factor may have contributed to the low prevalence of HIV infection in Finland. The stage and age of the epidemic should be taken into account when interpreting the data on late HIV diagnosis, especially in cross-country comparisons. PMID- 20002777 TI - Non-medically supervised treatment interruptions among participants in a universally accessible antiretroviral therapy programme. AB - BACKGROUND: We examined clinical outcomes, patient characteristics and trends over time of non-medically supervised treatment interruptions (TIs) from a free of-charge antiretroviral therapy (ART) programme in British Columbia (BC), Canada. METHODS: Data from ART-naive individuals > or =18 years old who initiated triple combination highly active antiretroviral therapy (HAART) between January 2000 and June 2006 were analysed. Participants having > or =3 month gap in HAART coverage were defined as having a TI. Cox proportional hazards modelling was used to examine factors associated with TIs and to examine factors associated with resumption of treatment. RESULTS: A total of 1707 participants were study eligible and 643 (37.7%) experienced TIs. TIs within 1 year of ART initiation decreased from 29% of individuals in 2000 to 19% in 2006 (P<0.001). TIs were independently associated with a history of injection drug use (IDU) (P=0.02), higher baseline CD4 cell counts (P<0.001), hepatitis C co-infection (P<0.001) and the use of nelfinavir (NFV) (P=0.04) or zidovudine (ZDV)/lamivudine (3TC) (P=0.009) in the primary HAART regimen. Male gender (P<0.001), older age (P<0.001), AIDS at baseline (P=0.008) and having a physician who had prescribed HAART to fewer patients (P=0.03) were protective against TIs. Four hundred and eighty-eight (71.9%) participants eventually restarted ART with male patients and those who developed an AIDS-defining illness prior to their TI more likely to restart therapy. Higher CD4 cell counts at the time of TI and unknown hepatitis C status were associated with a reduced likelihood of restarting ART. CONCLUSION: Treatment interruptions were associated with younger, less ill, female and IDU participants. Most participants with interruptions eventually restarted therapy. Interruptions occurred less frequently in recent years. PMID- 20002778 TI - Hospitalization risk following initiation of highly active antiretroviral therapy. AB - OBJECTIVES: While highly active antiretroviral therapy (HAART) decreases long term morbidity and mortality, its short-term effect on hospitalization rates is unknown. The primary objective of this study was to determine hospitalization rates over time in the year after HAART initiation for virological responders and nonresponders. METHODS: Hospitalizations among 1327 HAART-naive subjects in an urban HIV clinic in 1997-2007 were examined before and after HAART initiation. Hospitalization rates were stratified by virological responders (> or =1 log(10) decrease in HIV-1 RNA within 6 months after HAART initiation) and nonresponders. Causes were determined through International Classification of Diseases, 9th Revision (ICD-9) codes and chart review. Multivariate negative binomial regression was used to assess factors associated with hospitalization. RESULTS: During the first 45 days after HAART initiation, the hospitalization rate of responders was similar to their pre-HAART baseline rate [75.1 vs. 78.8/100 person years (PY)] and to the hospitalization rate of nonresponders during the first 45 days (79.4/100 PY). The hospitalization rate of responders fell significantly between 45 and 90 days after HAART initiation and reached a plateau at approximately 45/100 PY from 91 to 365 days after HAART initiation. Significant decreases were seen in hospitalizations for opportunistic and nonopportunistic infections. CONCLUSIONS: The first substantial clinical benefit from HAART may be realized by 90 days after HAART initiation; providers should keep close vigilance at least until this time. PMID- 20002779 TI - Long-term high-physiological-dose growth hormone reduces intra-abdominal fat in HIV-infected patients with a neutral effect on glucose metabolism. AB - OBJECTIVES: The aim of the study was to investigate the effect of long-term high physiological-dose recombinant human growth hormone (rhGH) therapy on fat distribution and glucose metabolism in HIV-infected patients. METHODS: Forty-six HIV-infected Caucasian men on highly active antiretroviral therapy (HAART), with an age range of 21-60 years and no significant comorbidity, were included in this randomized, placebo-controlled, double-blind, single-centre trial. Twenty-eight subjects were randomized to 0.7 mg/day rhGH, and 18 subjects to placebo, administered as daily subcutaneous injections between 1 and 3 pm for 40 weeks. Endpoints included changes in visceral adipose tissue (VAT), subcutaneous adipose tissue (SAT), limb fat mass, percentage of limb fat, plasma lipids, insulin resistance and glucose tolerance. RESULTS: VAT and trunk fat mass decreased significantly in the GH group compared with the placebo group [-19 cm(2) (-11%) vs. 12 cm(2) (6%), P=0.03, and -548 g (-9%) vs. 353 g (6%), P<0.01, respectively]. The beneficial fat redistribution in the GH group occurred without concomitant changes in subcutaneous fat at the abdomen or extremities. rhGH therapy was well tolerated. Insulin resistance, glucose tolerance, and total plasma cholesterol and triglycerides did not significantly change during intervention. CONCLUSIONS: Daily 0.7 mg rhGH treatment for 40 weeks reduced abdominal visceral fat and trunk fat mass in HIV-infected patients. This treatment appeared to be safe with respect to glucose tolerance and insulin sensitivity. PMID- 20002780 TI - Pitfalls in measuring high-density lipoprotein cholesterol concentrations in HIV infected patients. AB - OBJECTIVES: HIV infection and its treatment are associated with dyslipidaemia and increased risk of cardiovascular disease. Accurate high-density lipoprotein (HDL) cholesterol values are necessary for the management of these abnormalities, but current methods have not been properly assessed in these patients. The aim of this study was to assess in HIV-infected patients the consistency and accuracy of a synthetic polymer/detergent homogeneous assay used to measure HDL cholesterol concentrations and to evaluate the impact of storage. METHODS: HDL cholesterol was measured using a synthetic polymer/detergent homogeneous method in samples from HIV-infected patients and healthy subjects for each of the storage regimens: baseline, after 1 week at 4 degrees C, and after 12 months at -80 degrees C. The ultracentrifugation and precipitation assays were used for comparison. RESULTS: Three out of every 20 samples from HIV-infected patients had discrepant HDL cholesterol values with respect to the ultracentrifugation method. Overestimation was associated with high C-reactive protein concentrations and underestimation with plasma gamma-globulin concentrations, an effect that was amplified by any of the storage conditions tested. CONCLUSIONS: Caution is needed when using the synthetic polymer/detergent homogeneous method for direct measurement of HDL cholesterol concentrations in HIV-infected patients. This assay is of limited use in clinical trials in which frozen samples are analysed. PMID- 20002781 TI - Use of a prescription-based measure of antiretroviral therapy adherence to predict viral rebound in HIV-infected individuals with viral suppression. AB - OBJECTIVE: The aim of the study was to assess whether a simple, routinely available measure of antiretroviral therapy (ART) adherence predicts viral rebound at the next HIV viral load (VL) measurement in virally suppressed patients. METHODS: The analysis was performed on the Royal Free HIV Cohort, London, UK. Each 'drug coverage-viral load episode' (DCVL episode) was defined as a 6-month period immediately prior to a VL < or =50 HIV-1 RNA copies/mL (time zero), during which the patient had been continuously on HAART, with all measured VLs < or =50 copies/mL. The next VL after time-zero was used to assess whether VL rebound (defined as >200 copies/mL) had occurred. Drug coverage, our measure of adherence, was calculated as the proportion of days in the 6-month period covered by a valid prescription for at least three antiretroviral drugs. RESULTS: A total of 376 (2.4%) VL rebounds occurred in 15 660 DCVL episodes among 1632 patients. Drug coverage was 100% for 32% of episodes, 95-99% for 16% of episodes and < or =60% for 10% of episodes. The risk ratio of rebound associated with a 10% increase in drug coverage, adjusted for potential confounding variables, was 0.93 (95% confidence interval 0.88-0.98). CONCLUSIONS: Antiretroviral drug coverage assessed at the time of VL measurement in patients with undetectable VL is potentially clinically useful for predicting VL rebound at the next VL measurement. PMID- 20002782 TI - The psychological status of HIV-positive people and their psychosocial experiences in eastern China. AB - OBJECTIVES: The aim of the study was to investigate the psychological status and the psychosocial experiences of HIV-positive people using Symptom Check List 90 (SCL-90) in eastern China. METHODS: Two hundred and fourteen HIV-positive people and 200 controls were recruited to the study. Participants were given an anonymous questionnaire which included questions pertaining to demography, SCL-90 and psychosocial experiences. RESULTS: The mean subscale scores for SCL-90 in the HIV-positive group were all higher than those of the control group (P<0.001), especially for depression, anxiety, obsessive-compulsive disorder and hostility. Female HIV-positive individuals had significantly higher depression and anxiety scores (P<0.05) and more scores higher than 2.0 than male HIV-positive individuals. The average number of subscales with mean scores higher than 2.0 was 4.1 for female HIV-positive individuals and 3.7 for male HIV-positive individuals. The most common psychosocial experiences related to HIV infection were fear (36.9%) and helplessness (31.8%). 90.2% of HIV-positive people would not tell others about their disease because of fear of discrimination against family members (42.2%), exclusion by community members (26.9%) and abandonment (23.3%). Discrimination from acquaintances (38.8%) was a main stressor in the HIV positive individuals' daily life. Most members of HIV-positive individuals' communities expressed negative attitudes: alienation, coldness, aversion and fear. 38.3% of the HIV-positive participants reported that their family members had been discriminated against. CONCLUSIONS: The results demonstrate that HIV positive people in eastern China live in a negative psychosocial environment and suffer from psychological distress. It is necessary to provide psychological interventions for people living with AIDS and to educate community members in order to improve the psychosocial environment. PMID- 20002783 TI - Lopinavir protein binding in HIV-1-infected pregnant women. AB - BACKGROUND: Pregnancy may alter protein binding (PB) of highly bound protease inhibitors due to changes in plasma concentrations of albumin and alpha-1 acid glycoprotein (AAG). Small changes in PB can greatly impact the fraction of drug unbound (FU) exerting pharmacological effect. We report lopinavir (LPV) PB during third trimester (antepartum, AP) compared to > or =1.7 weeks postpartum (PP) to determine if FU changes compensate for reduced total concentrations reported previously. METHODS: P1026s enrolled women receiving LPV/ritonavir, soft gel capsules 400/100 mg or 533/133 mg twice daily. LPV FU, albumin and AAG were determined AP and PP. RESULTS: AP/PP samples were available from 29/25 women respectively with all but one woman receiving the same dose AP/PP. LPV FU was increased 18% AP vs. PP (mean 0.96+/-0.16% AP vs. 0.82+/-0.21% PP, P=0.001). Mean protein concentrations were reduced AP (AAG=477 mg/L; albumin=3.28 mg/dL) vs. PP (AAG=1007 mg/L; albumin=3.85 mg/dL) (P<0.0001 for each comparison). AAG concentration correlated with LPV binding. Total LPV concentration did not correlate with LPV FU AP or PP. However, higher LPV concentration PP was associated with reduced PB and higher FU after adjustment for AAG. CONCLUSIONS: LPV FU was higher and AAG lower AP vs. PP. The 18% increase in LPV FU AP is smaller than the reduction in total LPV concentration reported previously and is not of sufficient magnitude to eliminate the need for an increased dose during pregnancy. PMID- 20002784 TI - CCL19 is a specific ligand of the constitutively recycling atypical human chemokine receptor CRAM-B. AB - The human chemokine receptor CRAM (chemokine receptor on activated macrophages), encoded by the gene CCRL2, is a new candidate for the atypical chemokine receptor family that includes the receptors DARC, D6 and chemocentryx chemokine receptor (CCX-CKR). CRAM is maturation-stage-dependently expressed on human B lymphocytes and its surface expression is up-regulated upon short-term CCL5 exposure. Here, we demonstrate that the homeostatic chemokine CCL19 is a specific ligand for CRAM. In radioactive labelling studies CCL19 bound to CRAM-expressing cells with an affinity similar to the described binding of its other receptor CCR7. In contrast to the known CCL19/CCR7 ligand/receptor pair, CRAM stimulation by CCL19 did not result in typical chemokine-receptor-dependent cellular activation like calcium mobilization or migration. Instead, we demonstrate that CRAM is constitutively recycling via clathrin-coated pits and able to internalize CCL19 as well as anti-CRAM antibodies. As this absence of classical chemokine receptor responses and the recycling and internalization features are characteristic for non-classical chemokine receptors, we suggest that CRAM is the newest member of this group. As CCL19 is known to be critically involved in lymphocyte and dendritic cell trafficking, CCL19-binding competition by CRAM might be involved in modulating these processes. PMID- 20002785 TI - Suboptimal engagement of the T-cell receptor by a variety of peptide-MHC ligands triggers T-cell anergy. AB - T cells recognize antigen via the T-cell receptor (TCR) and produce a spectrum of responses that range from activation to anergy or cell death. The variety of outcomes may be dictated by the strength of the signals transmitted upon cognate recognition of the TCR. The physiological outcome of TCR engagement is determined by several factors, including the avidity of the ligand for TCR, the duration of engagement, and the presence and nature of accessory molecules present on antigen presenting cells (APCs). In this review, we discuss a model of anergy induced by presentation of low densities of peptide-major histocompatibility complex (MHC) ligand in CD4(+) T cells and compare it to anergy induced by altered peptide ligands in an effort to identify a unifying mechanism. We suggest that altered peptide ligand (APL) and low densities of agonist ligands induce anergy by engaging less than optimal numbers of TCRs. The physiological impacts of anergy in memory CD4(+) T cells are discussed. PMID- 20002786 TI - Hypoxic stress up-regulates the expression of Toll-like receptor 4 in macrophages via hypoxia-inducible factor. AB - Toll-like receptors (TLRs) are germline-encoded innate immune receptors that recognize invading micro-organisms and induce immune and inflammatory responses. Deregulation of TLRs is known to be closely linked to various immune disorders and inflammatory diseases. Cells at sites of inflammation are exposed to hypoxic stress, which further aggravates inflammatory processes. We have examined if hypoxic stress modulates the TLR activity of macrophages. Hypoxia and CoCl(2) (a hypoxia mimetic) enhanced the expression of TLR4 messenger RNA and protein in macrophages (RAW264.7 cells), whereas the messenger RNA of other TLRs was not increased. To determine the underlying mechanism, we investigated the role of hypoxia-inducible factor 1 (HIF-1) in the regulation of TLR4 expression. Knockdown of HIF-1alpha expression by small interfering RNA inhibited hypoxia induced and CoCl(2)-induced TLR4 expression in macrophages, while over-expression of HIF-1alpha potentiated TLR4 expression. Chromatin immunoprecipitation assays revealed that HIF-1alpha binds to the TLR4 promoter region under hypoxic conditions. In addition, deletion or mutation of a putative HIF-1-binding motif in the TLR4 promoter greatly attenuated HIF-1alpha-induced TLR4 promoter reporter expression. Up-regulation of TLR4 expression by hypoxic stress enhanced the response of macrophages to lipopolysaccharide, resulting in increased expression of cyclooxygenase-2, interleukin-6, regulated on activation normal T cell expressed and secreted, and interferon-inducible protein-10. These results demonstrate that TLR4 expression in macrophages is up-regulated via HIF-1 in response to hypoxic stress, suggesting that hypoxic stress at sites of inflammation enhances susceptibility to subsequent infection and inflammatory signals by up-regulating TLR4. PMID- 20002787 TI - Activation of mannan-binding lectin-associated serine proteases leads to generation of a fibrin clot. AB - The lectin pathway of complement is activated upon binding of mannan-binding lectin (MBL) or ficolins (FCNs) to their targets. Upon recognition of targets, the MBL-and FCN-associated serine proteases (MASPs) are activated, allowing them to generate the C3 convertase C4b2a. Recent findings indicate that the MASPs also activate components of the coagulation system. We have previously shown that MASP 1 has thrombin-like activity whereby it cleaves and activates fibrinogen and factor XIII. MASP-2 has factor Xa-like activity and activates prothrombin through cleavage to form thrombin. We now report that purified L-FCN-MASPs complexes, bound from serum to N-acetylcysteine-Sepharose, or MBL-MASPs complexes, bound to mannan-agarose, generate clots when incubated with calcified plasma or purified fibrinogen and factor XIII. Plasmin digestion of the clot and analysis using anti D-dimer antibodies revealed that the clot was made up of fibrin and was similar to that generated by thrombin in normal human plasma. Fibrinopeptides A and B (FPA and FPB, respectively) were released after fibrinogen cleavage by L-FCN MASPs complexes captured on N-acetylcysteine-Sepharose. Studies of inhibition of fibrinopeptide release indicated that the dominant pathway for clotting catalysed by the MASPs is via MASP-2 and prothrombin activation, as hirudin, a thrombin inhibitor that does not inhibit MASP-1 and MASP-2, substantially inhibits fibrinopeptide release. In the light of their potent chemoattractant effects on neutrophil and fibroblast recruitment, the MASP-mediated release of FPA and FPB may play a role in early immune activation. Additionally, MASP-catalysed deposition and polymerization of fibrin on the surface of micro-organisms may be protective by limiting the dissemination of infection. PMID- 20002788 TI - Antigenic dietary protein guides maturation of the host immune system promoting resistance to Leishmania major infection in C57BL/6 mice. AB - The immature immune system requires constant stimulation by foreign antigens during the early stages of life to develop properly and to create efficient immune responses against later infections. We have previously shown that intake of antigenic dietary protein is critical for inducing maturation of the immune system as well as for the development of T helper type 1 (Th1) immunity. In this study, we show that administration of an amino acid (aa)-based diet during the development of the immune system subsequently resulted in inefficient control of Leishmania major infection in adult C57BL/6 mice. Compared with mice fed a control protein-containing diet, adult aa-fed mice showed a decreased interferon (IFN)-gamma response to parasite antigens and insufficient production of nitric oxide (NO), which is crucial to parasite death. However, no deviation towards Th2 specific immunity to L. major was observed. Phenotypic analysis of antigen presenting cells (APCs) from aa-fed mice revealed deficient levels of the costimulatory molecules CD40 and CD80, and low levels of interleukin (IL)-12 produced by peritoneal macrophages, revealing an early stage of maturation of these cells. APCs isolated from aa-fed mice were unable to stimulate a Th1 response in vitro. Both phenotypic features of T cells from aa-fed mice and their ability to produce a Th1 response in the presence of mature APCs were unaffected when compared with T cells from control mice. The results presented here support the notion that regulation of Th1 immunity to infection includes environmental factors such as dietary proteins, which provide a natural source of stimulation that contributes to the process of maturation of APCs. PMID- 20002789 TI - Human immunodeficiency virus (HIV) infection during pregnancy induces CD4 T-cell differentiation and modulates responses to Bacille Calmette-Guerin (BCG) vaccine in HIV-uninfected infants. AB - Human immunodeficiency virus (HIV)-negative infants born to HIV-positive mothers frequently exhibit a range of immunological abnormalities. We tested the hypothesis that HIV during pregnancy affects the ability of CD4 T cells of HIV negative infants to respond to vaccine challenge by recruiting HIV-negative infants born to HIV-negative and HIV-positive mothers and measuring their responses to Bacille Calmette-Guerin (BCG) vaccine given at birth. At 2 weeks, maternal HIV status did not influence CD4 T-cell counts or differentiation, but by 10 weeks CD4 counts of infants born to HIV-positive mothers fell to a level characteristic of HIV-positive infants. Among the CD4 T-cell populations, markers of differentiation (CCR7(-) CD45RA(-) CD27(-)) and senescence (CD57, PD-1) were more common among infants born to HIV-positive mothers than among infants born to HIV-negative mothers. At 2 weeks of age, we assessed the effector response to heat-killed BCG and tuberculin purified protein derivative (PPD) by overnight interferon (IFN)-gamma enzyme-linked immunosorbent spot-forming cell assay (ELISpot), but found no measurable effect of maternal HIV status. At 10 weeks, we assessed CD4 T-cell memory by measuring proliferation in response to the same antigens. We observed a bimodal response that allowed infants to be classified as high or low responders and found that fewer infants born to HIV-positive mothers were able to mount a robust proliferative response, suggesting that their reduced CD4 counts and increased differentiation indicated a deficiency in their ability to develop immunological memory. PMID- 20002790 TI - The expression and function of Nod-like receptors in neutrophils. AB - Neutrophils make up an essential part of the innate immune system, and are involved both in the initial responses to pathogens, and in orchestrating later immune responses. Neutrophils recognize pathogens through pattern-recognition receptors (PRRs), which are activated by microbial motifs. The Nod-like receptors (nucleotide-binding domain leucine-rich repeat containing family; NLRs) constitute a recently discovered group of PRRs whose role in the neutrophil immune responses is not yet characterized. The present study aimed to investigate the expression and function of NLRs in neutrophils. Neutrophils were isolated from human peripheral blood, and the presence of nucleotide-binding oligomerization domain 1 (NOD1), NOD2 and NACHT-LRR-PYD-containing protein 3 (NLRP3) was evaluated with flow cytometry and immunohistochemistry. The expression of NOD1, NOD2 and NLRP3 messenger RNA was determined using real-time reverse transcription-polymerase chain reaction. Changes in neutrophil cytokine secretion, phenotype and migration following agonist-induced activation were studied using enzyme-linked immunosorbent assay, flow cytometry and a chemotaxis assay, respectively. No expression of NOD1 was found in isolated neutrophils and stimulation with the NOD1 ligand gamma-d-glutamyl-meso-diaminopimelic acid induced no signs of activity. In contrast, a marked expression of NOD2 and NLRP3 was found. NOD2 activation with MurNAc-l-Ala-d-isoGln (MDP) resulted in interleukin-8 secretion, CD62 ligand down-regulation, CD11b up-regulation and increased migration towards an inflammatory stimulus. NLRP3 activation with alum caused interleukin-1beta secretion and facilitated migration. Altogether, this suggests that NLRs may be a previously unknown pathway for neutrophil activation. PMID- 20002792 TI - Some questions on dispersion of human exhaled droplets in ventilation room: answers from numerical investigation. AB - This study employs a numerical model to investigate the dispersion characteristics of human exhaled droplets in ventilation rooms. The numerical model is validated by two different experiments prior to the application for the studied cases. Some typical questions on studying dispersion of human exhaled droplets indoors are reviewed and numerical study using the normalized evaporation time and normalized gravitational sedimentation time was performed to obtain the answers. It was found that modeling the transient process from a droplet to a droplet nucleus due to evaporation can be neglected when the normalized evaporation time is <0.051. When the normalized gravitational sedimentation time is <0.005, the influence of ventilation rate could be neglected. However, the influence of ventilation pattern and initial exhaled velocity on the exhaled droplets dispersion is dominant as the airflow decides the droplets dispersion significantly. Besides, the influence of temperature and relative humidity on the dispersion of droplets can be neglected for the droplet with initial diameter <200 microm; while droplet nuclei size plays an important role only for the droplets with initial diameter within the range of 10 microm 100 microm. Practical Implications Dispersion of human exhaled droplets indoor is a key issue when evaluating human exposure to infectious droplets. Results from detailed numerical studies in this study reveal how the evaporation of droplets, ventilation rate, airflow pattern, initial exhaled velocity, and particle component decide the droplet dispersion indoor. The detailed analysis of these main influencing factors on droplet dispersion in ventilation rooms may help to guide (1) the selection of numerical approach, e.g., if the transient process from a droplet to a droplet nucleus due to evaporation should be incorporated to study droplet dispersion, and (2) the selection of ventilation system to minimize the spread of pathogen-laden droplets in an indoor environment. PMID- 20002791 TI - Anti-Aspergillus human host defence relies on type 1 T helper (Th1), rather than type 17 T helper (Th17), cellular immunity. AB - Both interferon-gamma-producing type 1 T helper (Th1)- and interleukin-17 (IL-17) producing Th17 cells have been proposed to be involved in anti-fungal host defence. Although invasive aspergillosis is one of the most severe human fungal infections, little is known regarding the relative importance of the Th1 versus Th17 cellular immune pathways for the human anti-Aspergillus host defence. Using human peripheral blood mononuclear cells and a system consisting of monocyte derived macrophages with lymphocytes, we found that Aspergillus fumigatus is a weak inducer of human IL-17 but induces a strong Th1 response. These data were validated by the very low IL-17 levels in bronchoalveolar lavage fluid and serum of patients with invasive aspergillosis. Surprisingly, live A. fumigatus reduced IL-17 production induced by mitogenic stimuli. This effect was mediated through the propensity of A. fumigatus to metabolize tryptophan and release kynurenine, which modulates the inflammatory response through inhibition of IL-17 production. In conclusion, A. fumigatus does not stimulate production of IL-17 and human host defence against aspergillosis may not rely on potent Th17 responses. PMID- 20002793 TI - Indoor/outdoor concentrations and elemental composition of PM10/PM2.5 in urban/industrial areas of Kocaeli City, Turkey. AB - This study presents indoor/outdoor PM2.5 and PM10 concentrations measured during winter and summer in 15 homes in Kocaeli, which is one of the most industrialized areas in Turkey. Indoor and outdoor PM2.5 and PM10 mass concentrations and elemental composition were determined using an X-ray fluorescence spectrometer. Quantitative information was obtained on mass concentrations and other characteristics such as seasonal variation, indoor/outdoor (I/O) ratio, PM2.5/PM10 ratio, correlations and sources. Average indoor and outdoor PM2.5 concentrations were 29.8 and 23.5 microg/m(3) for the summer period, and 24.4 and 21.8 microg/m(3) for the winter period, respectively. Average indoor and outdoor PM10 concentrations were 45.5 and 59.9 microg/m(3) for the summer period, and 56.9 and 102.3 microg/m(3) for the winter period, respectively. A varimax rotated factor analysis (FA) was performed separately on indoor and outdoor datasets in an effort to identify possible heavy metal sources of PM2.5 and PM10 particle fractions. FA of outdoor data produced source categories comprising polluted soil, industry, motor vehicles, and fossil fuel combustion for both PM fractions, while source categories determined for indoor data for both PM2.5 and PM10 comprised industry, polluted soil, motor vehicles, and smoking, with an additional source category of cooking activities detected for the PM2.5 fraction. Practical Implications In buildings close to industrial areas or traffic arteries, outdoor sources may have an important effect on indoor air pollution. Therefore, indoor and outdoor investigations should be conducted simultaneously to assess the relationship between indoor and outdoor pollution. This study presents the simultaneous measurement of PM fractions (PM2.5 and PM10) and their elemental compositions to determine the sources of respirable PM and the heavy metals bound to these particles in indoor air. Factor analysis of indoor data indicated that the contribution of outdoor pollutant sources to indoor pollution was about 70%, making these sources the most significant for indoor heavy metal pollution, wheras other sources of indoor pollution included smoking and cooking activities. PMID- 20002794 TI - Short-term airing by natural ventilation - implication on IAQ and thermal comfort. AB - The need to improve the energy efficiency of buildings requires new and more efficient ventilation systems. It has been demonstrated that innovative operating concepts that make use of natural ventilation seem to be more appreciated by occupants. Among the available ventilation strategies that are currently available, buoyancy driven, single-sided natural ventilation has proved to be very effective and can provide high air change rates for temperature and Indoor Air Quality (IAQ) control. However, to promote a wider distribution of these systems an improvement in the knowledge of their working principles is necessary. The present study analyses and presents the results of an experimental evaluation of airing performance in terms of ventilation characteristics, IAQ and thermal comfort. It includes investigations of the consequences of opening time, opening frequency, opening area and expected airflow rate, ventilation efficiency, thermal comfort and dynamic temperature conditions. A suitable laboratory test rig was developed to perform extensive experimental analyses of the phenomenon under controlled and repeatable conditions. The results showed that short-term window airing is very effective and can provide both acceptable IAQ and thermal comfort conditions in buildings. Practical Implications This study gives the necessary background and in-depth knowledge of the performance of window airing by single-sided natural ventilation necessary for the development of control strategies for window airing (length of opening period and opening frequency) for optimum IAQ and thermal comfort in naturally ventilated buildings. PMID- 20002795 TI - Silkmoth chorion gene regulation revisited: promoter architecture as a key player. AB - Regulation of silkmoth chorion genes has long been used as a model system for studying differential gene expression. The large numbers of genes, their overlapping expression patterns and the overall complexity of the system hinted towards an elaborate mechanism for transcriptional control. Recent studies, however, offer evidence of a molecular pathway governed by the interplay between two general transcription factors, CCAAT enhancer binding proteins (C/EBP) and GATA, an architectural protein, high mobility group A and a chromatin remodeller, chromo-helicase/ATPase-DNA binding protein 1. In this review we present a parsimonious model that adequately describes regulation of transcription across all temporally regulated chorion genes, and propose a role for promoter architecture. PMID- 20002796 TI - Transcriptome analysis of the synganglion from the brown dog tick, Rhipicephalus sanguineus. AB - Tick control strategies rely heavily on chemicals (acaricides), most of which target the central nervous system. With increasing resistance, new acaricides are urgently needed but knowledge of tick neurobiology is surprisingly limited, notably the number of neural-specific gene sequences. One thousand and eight expressed sequence tags (ESTs) were obtained from a normalized cDNA library from Rhipicephalus sanguineus synganglia. Putative functional identities were assigned to 44% whereas 34% were unknown/novel sequences. Of particular interest were ESTs encoding a chitinase-like enzyme, an acetylcholinesterase and four transmembrane receptors including two glutamate-gated chloride channel receptors, a leucokinin like receptor and a nicotinic acetylcholine receptor alpha-subunit. This study highlights the benefits of using both neural tissues and normalized libraries in an EST-approach for identifying potential acaricide targets expressed as rare transcripts. PMID- 20002797 TI - A cathepsin L-like proteinase is involved in moulting and metamorphosis in Helicoverpa armigera. AB - Holometabolous insects undergo larval moulting and metamorphosis within their life cycle. A cDNA encoding the cathepsin L-like proteinase Ha-cathL has been cloned from Helicoverpa armigera. It has a sequence of 1826 bp and encodes a 550 residue protein with a molecular mass of 63 kDa. Northern blot analysis indicated that Ha-cathL is specifically expressed in haemocytes, with increased expression during larval moulting and metamorphosis. In vivo experimentation revealed that Ha-cathL is up-regulated by 20-hydroxyecdysone. Meanwhile, in situ hybridization and immunocytochemistry revealed that Ha-cathL mRNA is mainly expressed in granulocytes and plasmatocytes. Knock down of cathepsin L by RNA interference results in larvae death before pupation or the formation of a chimeric pupa containing a larval head and thorax, abnormal wings and the pupal abdomen. The reason for this is that the affected haemocytes cannot become granulated, and therefore cannot participate in fat body remodelling and wing development. These facts suggest that Ha-cathL is involved in larval moulting and metamorphosis by participating in the functioning of haemocytes. PMID- 20002799 TI - The smear layer in endodontics - a review. AB - Root canal instrumentation produces a layer of organic and inorganic material called the smear layer that may also contain bacteria and their by-products. It can prevent the penetration of intracanal medicaments into dentinal tubules and influence the adaptation of filling materials to canal walls. This article provides an overview of the smear layer, focusing on its relevance to endodontics. The PubMed database was used initially; the reference list for smear layer featured 1277 articles, and for both smear layer dentine and smear layer root canal revealed 1455 publications. Smear layer endodontics disclosed 408 papers. A forward search was undertaken on selected articles and using some author names. Potentially relevant material was also sought in contemporary endodontic texts, whilst older books revealed historic information and primary research not found electronically, such that this paper does not represent a 'classical' review. Data obtained suggests that smear layer removal should enhance canal disinfection. Current methods of smear removal include chemical, ultrasonic and laser techniques - none of which are totally effective throughout the length of all canals or are universally accepted. If smear is to be removed, the method of choice seems to be the alternate use of ethylenediaminetetraacetic acid and sodium hypochlorite solutions. Conflict remains regarding the removal of the smear layer before filling root canals, with investigations required to determine the role of the smear layer in the outcomes of root canal treatment. PMID- 20002800 TI - Comparison of working length determination with radiographs and two electronic apex locators. AB - AIM: To evaluate the accuracy of the Root ZX and Elements-Diagnostic electronic apex locators when compared with radiographs for locating the canal terminus or minor foramen. METHODOLOGY: The canal terminus of 482 canals in 160 maxillary and mandibular teeth was located in vivo with both locators and radiographically. After extraction, the actual location of the minor foramen was determined visually and with magnification. A paired samples t-test, chi-square test and a repeated measure anova at the 0.05 level of significance were used to determine differences between the groups. RESULTS: The Root ZX located the minor foramen correctly 68% of the time in anterior and premolar teeth, and 58% of the time in molar teeth. The Elements-Diagnostic located the minor foramen correctly 58% of the time in anterior and premolar teeth and 49% of the time in molar teeth. Radiographs located the minor foramen correctly 20% of the time in anterior and premolar teeth and 11% of the time in molar teeth. There was no statistically significant difference between the two locators, but there was a significant difference between them and radiographs. For all teeth, the measurements made by the apex locators were within +/-0.5 mm of the minor foramen 100% of the time, whereas for the radiographs, the measurements were within this range only 15% of the time. This difference was significant (P = 0.05). CONCLUSION: Measuring the location of the minor foramen using the two apex locators was more accurate than radiographs and would reduce the risk of instrumenting and filling beyond the apical foramen. PMID- 20002801 TI - The effect of sodium hypochlorite and ethylenediaminetetraacetic acid irrigation, individually and in alternation, on tooth surface strain. AB - AIM: To evaluate the effect of irrigation regimens on tooth surface strain using saline, sodium hypochlorite (3% and 5% NaOCl) and ethylenediaminetetraacetic acid (17% EDTA), individually and in alternating combinations. METHODOLOGY: Single rooted premolar teeth with single canals prepared to standardized dimensions were grouped by anatomical features and randomly distributed amongst six experimental groups (n = 12 each). The six groups were: (1) saline; (2) 5% NaOCl; (3) 3% NaOCl; (4) 17% EDTA; (5) 3% NaOCl and 17% EDTA; (6) 5% NaOCl and 17% EDTA. All groups underwent four (group 1) or five (groups 2, 3, 4, 5, 6) sequential 30-min irrigation periods following each of which the tooth was subjected to a standard regime of cyclic, nondestructive, occlusal loading. Tooth surface strain was measured during each loading cycle using electrical strain gauges mounted cervico proximally. The data were analysed by Hierarchical anova and post hoc multiple comparisons. RESULTS: Irrigation with 5% NaOCl alone or alternating with 17% EDTA significantly (P < 0.001) increased the peak strain values for each of the irrigation periods compared with that of saline (group 1). The data for the other groups revealed no significant differences compared with those of saline. The strain increase after the fourth irrigation cycle was significantly higher for group 6 than for group 2. The measured canal morphology and dentine thickness parameters did not prove to have a significant effect on tooth surface strain. CONCLUSIONS: Irrigation with 5% NaOCl acting alone or alternated with 17% EDTA (used in 30 min cycles), significantly increased tooth surface strain. The alternated regimen showed significantly greater changes in tooth surface strain than NaOCl alone. Irrigation with 3% NaOCl and 17% EDTA individually or in combination did not significantly alter the tooth surface strain. PMID- 20002802 TI - Analysis of resin tags formation in root canal dentine: a cross sectional study. AB - AIM: To evaluate the length, density and quality of resin tags formed by penetration of various types of adhesive systems into dentinal tubules at various cross section levels of the root canal in correlation to the density of dentinal tubules. METHODOLOGY: Thirty mandibular premolars were instrumented and fibre posts were inserted with three different adhesive systems with and without activator: etch & rinse XP Bond and XP Bond/Self Cure Activator; self-etch (two step) AdheSE and AdheSE/AdheSE DC Activator and self-etch (one-step) Hybrid Bond and Hybrid Bond/Hybrid Brushes. The resin tags were evaluated from slices obtained from sections perpendicular to the long axis of the teeth at 3, 6, and 9 mm from the root apex under a Confocal Laser Scanning microscope. RESULTS: In all groups, lack of continuity of resin tag length, density and quality was observed not only from the cervical to the apical region of each root canal, but also in a mesio-distal direction to the long axis of the root. Application of etch & rinse adhesive in contrast to the self-etch adhesives provided the formation of the shorter, but considerably denser, more homogeneous and not interrupted resin tags with similar length. Use of the activator for all types of adhesives significantly increased the completeness (P = 0.014) and continuity (P = 0.024) of resin tags. CONCLUSIONS: None of the investigated adhesives were able to completely infiltrate the dentinal tubules in the entire root canal. Use of the etch & rinse adhesive system and the activators significantly increased the density and the quality of resin tags. PMID- 20002803 TI - Haemostatic effect and tissue reactions of methods and agents used for haemorrhage control in apical surgery. AB - AIM: To compare the haemostatic effect and tissue reactions of different agents and methods used for haemorrhage control in apical surgery. METHODOLOGY: Six standardized bone defects were prepared in the calvaria of six Burgundy rabbits. Five haemostatic modalities were tested for their haemostatic effect and tissue reactions, and were compared with untreated control defects: Expasyl + Stasis, Expasyl + Stasi + freshening of the bone defect with a bur, Spongostan, Spongostan+ epinephrine, and electro cauterization. The haemostatic effect was analysed visually and compared using Wilcoxon's signed rank test. Two groups of three animals were evaluated histologically for hard and soft tissue reactions related to the different haemostatic measures, after 3 and 12 weeks of healing respectively. RESULTS: Expasyl + Stasis and electro cauterization proved most effective in reducing bleeding (P < 0.05), but were accompanied by unfavourable tissue reactions, as indicated by the presence of necrotic bone, inflammatory cells and the absence of bone repair. These adverse tissue reactions did not recover substantially over time. However, adverse reactions were not observed when the superficial layer of bone had been removed with a rotary instrument. In contrast, Spongostan + epinephrine showed only a moderate haemostatic effect, but elicited also only mild adverse tissue reactions. CONCLUSIONS: Haemostasis in experimental bone defects is most effectively accomplished by using Expasyl + Stasis or electro cauterization. However, the bone defects should be freshened with a rotary instrument before suturing so as not to compromise healing. PMID- 20002804 TI - Apexogenesis after initial root canal treatment of an immature maxillary incisor a case report. AB - AIM: To present a case where a traumatized, immature tooth still showed capacity for continued root development and apexogenesis after root canal treatment was initiated based on an inaccurate pulpal diagnosis. SUMMARY: Traumatic dental injuries may result in endodontic complications. Treatment strategies for traumatized, immature teeth should aim at preserving pulp vitality to ensure further root development and tooth maturation. A 9-year-old boy, who had suffered a concussion injury to the maxillary anterior teeth, was referred after endodontic treatment was initiated in tooth 21 one week earlier. The tooth had incomplete root length, thin dentinal walls and a wide open apex. The pulp chamber had been accessed, and the pulp canal instrumented to size 100. According to the referral, bleeding from the root made it difficult to fill the root canal with calcium hydroxide. No radiographic signs of apical breakdown were recorded. Based on radiographic and clinical findings, a conservative treatment approach was followed to allow continued root development. Follow-up with radiographic examination every 3rd month was performed for 15 months. Continued root formation with apical closure was recorded. In the cervical area, a hard tissue barrier developed, which was sealed with white mineral trioxide aggregate (MTA). Bonded composite was used to seal the access cavity. At the final 2 years follow-up, the tooth showed further root development and was free from symptoms. PMID- 20002805 TI - Non-resolving periapical inflammation: a malignant deception. AB - AIM: To report a case of oral non-Hodgkin's lymphoma with a delayed diagnosis. SUMMARY: Non-Hodgkin's lymphoma of the oral cavity is an uncommon but important condition. Early diagnosis is complicated when the presenting signs and symptoms are similar to those of odontogenic infections. This report describes the case of a 38-year-old female patient who presented to her dentist complaining of pain in her upper jaw. Subsequent dental treatment, including extraction, root canal treatment and apicectomy including biopsy were carried out by the patient's dentist and local dental hospital. Nine months elapsed before a more extensive surgical exploration established a diagnosis of lymphoma. PMID- 20002807 TI - Selectivity of lynx proteins on insect nicotinic acetylcholine receptors in the brown planthopper, Nilaparvata lugens. AB - Neuronal nicotinic acetylcholine receptors (nAChRs) are major excitatory neurotransmitter receptors in both vertebrates and invertebrates. Two lynx proteins (Nl-lynx1 and Nl-lynx2) have been identified in the brown planthopper, Nilaparvata lugens, which act as modulators on insect nAChRs. In the present study, two lynx proteins were found to act on the triplet receptor Nlalpha1/Nlalpha2/beta2 expressed in Xenopus oocytes, increasing agonist-evoked macroscopic currents, but not changing agonist sensitivity and desensitization properties. Nl-lynx1 and Nl-lynx2 increased I(max) (maximum responses) of acetylcholine to 4.85-fold and 2.40-fold of that of Nlalpha1/Nlalpha2/beta2 alone, and they also increased I(max) of imidacloprid to 2.57-fold and 1.25-fold. Although, on another triplet nAChRs Nlalpha3/Nlalpha8/beta2, Nl-lynx2 increased I(max) of acetylcholine and imidacloprid to 3.63-fold and 2.16-fold, Nl-lynx1 had no effects on I(max) of either acetylcholine or imidacloprid. The results demonstrate the selectivity of lynx proteins for different insect nAChR subtypes. This selectivity was also identified in native N. Lugens. Co-immunoprecipitation was found between Nlalpha1/Nlalpha2-containing receptors and both Nl-lynx1 and Nl lynx2, but was only found between Nlalpha3/Nlalpha8-containing receptors and Nl lynx2. When the previously identified Nlalpha1(Y151S) and Nlalpha3(Y151S) mutations were included (Nlalpha1(Y151S)/Nlalpha2/beta2 and Nlalpha3(Y151S)/Nlalpha8/beta2), the increase in I(max) of imidacloprid, but not acetylcholine, caused by co-expression of Nl-lynx1 and Nl-lynx2 was more noticeable than that of their wildtype counterparts. Taken together, these data suggest that two modulators, Nl-lynx1 and Nl-lynx2, might serve as an influencing factor in target site insensitivity in N. lugens, such as Y151S mutation. PMID- 20002809 TI - Toll-like receptors 2 and 4 gene polymorphisms in a southeastern Chinese population with tuberculosis. AB - Polymorphisms of the toll-like receptor 2 (TLR2) gene (Arg677Trp, Arg753Gln) and the TLR4 gene (Asp299Gly, Thr399Ile) were investigated in 205 tuberculosis (TB) patients and 203 controls. Genetic variations were analysed by DNA sequencing. We revealed that these polymorphisms were rare in the southeastern Chinese population and were not associated with susceptibility to TB. PMID- 20002808 TI - The effects of mating and instrumental insemination on queen honey bee flight behaviour and gene expression. AB - Mating is fundamental to most organisms, although the physiological and transcriptional changes associated with this process have been largely characterized only in Drosophila melanogaster. In this study, we use honey bees as a model system because their queens undergo massive and permanent physiological and behavioural changes following mating. Previous studies have identified changes associated with the transition from a virgin queen to a fully mated, egg-laying queen. Here, we further uncouple the mating process to examine the effects of natural mating vs. instrumental insemination and saline vs. semen insemination. We observed effects on flight behaviour, vitellogenin expression and significant overlap in transcriptional profiles between our study and analogous studies in D. melanogaster, suggesting that some post-mating mechanisms are conserved across insect orders. PMID- 20002810 TI - Study on VNTR polymorphism of gene IL-1RA in 19 Chinese populations. AB - Earlier studies suggested that a variable number tandem repeat (VNTR) polymorphism in intron 2 of the interleukin-1 receptor antagonist (IL-1RA) gene might be associated with some chronic inflammatory diseases, autoimmune diseases and solid tumours. To study the distribution of this polymorphism in China, 1352 samples were collected from 19 widely distributed Chinese populations. PCR was used to genotype the VNTR. The overall frequencies of allele 1 and allele 2 were 0.913 and 0.064 respectively. The frequency of the allele 2 was significantly different between the northeastern and the northwestern populations. Moreover, the allele frequencies at this locus in three Chinese Han populations were different from that in minority populations. When compared with other populations worldwide, the frequencies of the two alleles in China were not significantly different from those in the Asian and Pacific Islands. However, the prevalence of allele 1 in China was significantly higher, and the prevalence of allele 2 was significantly lower, than those in American and European Caucasians, and the pairwise Fst values reinforced this observation. The differences of the allele frequencies between different regions and within the same region showed that geography and race have important roles in the population differentiation for the IL-1RA gene. In summary, our results provide a valuable reference for population genetic information and future disease association studies in Chinese populations. PMID- 20002811 TI - Immunity-related gene single nucleotide polymorphisms associated with Rhodococcus equi infection in foals. AB - In previous work, we found significant associations of horse polymorphic microsatellite and immunity-related (IR) gene markers with Rhodococcus equi infection of foals. Here, a statistically significant association between a single nucleotide polymorphism (SNP) within the interleukin 7 receptor-encoding gene (IL7R) with high R. equi burden in transtracheal aspirates was found (Fisher's F = 0.043, odds ratio: 8.00, 95% confidence interval: 1.127-56.795). Further positional and/or functional candidate genes investigated TLR2, IL13, IL17A, IL28R, TACE/ADAM 17 and GBP1, were not associated with infection in this study. SNPs analysed were found by sequencing and appropriate restriction fragment length polymorphism markers were developed. Their associations with R. equi infection were tested by genotyping thoroughbred foals from the original study. The association was confirmed by analysing genotypes composed with genes previously reported to be associated with R. equi infection in the same group. PMID- 20002813 TI - Trauma theory in Sandor Ferenczi's writings of 1931 and 1932. AB - The author states that it is Ferenczi 's writings of 1931 and 1932 that exhibit the most conspicuous departures from Freud 's ideas and at the same time contain Ferenczi 's most original contributions. The texts concerned - Confusion of tongues between adults and the child (Ferenczi, 1932a), the Clinical Diary (Dupont, 1985), and some of the Notes and fragments (Ferenczi, 1930-32), all of which were published posthumously - present valuable and original theories on trauma which are significant not only in historical terms but also because the ideas concerned are relevant to our conception of clinical psychoanalysis today. The aim of this paper is to give an account of Ferenczi 's trauma theory as it emerges from his writings of 1931-32 and to specify the points on which he differs from Freud. PMID- 20002814 TI - The presence of Spinoza in the exchanges between Sigmund Freud and Romain Rolland. AB - Although Freud recognized his profound affinity with Spinoza, we seldom find explicit and direct references to the philosopher in his works. The correspondence between Romain Rolland, the 'Christian without a church', and Freud, the 'atheist Jew', is full of Spinozian reminiscences that nourish their works of this period and are underpinned by their mutual transference. The Future of an Illusion is written according to a Spinozian blueprint and aims at replacing religion, qualified as superstition, by psychoanalysis. A quotation from Heine, 'brother in unbelief', is a direct reference to Spinoza. Concurring with Freud's critiques of dogmas and churches, Rolland proposes an analysis of the 'oceanic feeling' as a basis of the religious sentiment. Freud replies with Civilization and Its Discontents. In 1936, on the occasion of Rolland's 70th birthday, Freud sends him an open letter, A disturbance of memory on the Acropolis, where the strange feeling that he has experienced in front of the Parthenon refers inter alia to his double culture: Jewish and German. In the light of this correspondence, the creation of psychoanalysis turns out to be a quest for the sacred that has disappeared in modernity; Freud, though, was able to find it inside man's unconscious. PMID- 20002815 TI - Child versus adult psychoanalysis: two processes or one? AB - Child analysis continues to be seen as a different technique from adult analysis because children are still involved in a developmental process and because the primary objects continue to play active roles in their lives. This paper argues that this is a false dichotomy. An extended vignette of the analysis of a latency aged girl is used to demonstrate that the psychoanalytic process that develops in child analysis is structurally the same as that in adult analysis. Both revolve around the analysis of resistance and transference and use both to promote knowledge of the patient's mind at work. And both techniques formulate interventions based on the analyst's appraisal of the patient's mental organization. It is hoped that stressing the essential commonality of both techniques will promote the development of an overarching theory of psychoanalytic technique. PMID- 20002816 TI - The relationship of the inner and the outer in psychoanalysis. AB - What is internal and what is external according to psychoanalytic theory? This is a surprisingly complicated question. The terminology is often ambiguous and inconsistent as, for instance, in the use of terms like 'object' and 'other'. The relationship between internal and external in psychoanalysis is analysed from a philosophical, concept analytical, developmental psychological, methodological and trauma versus internal dynamics point of view. It is argued that psychoanalytical writing is influenced by the authors' need to create their personal psychoanalytic theory and language. This is seen as one of the main reasons for the terminological variety and ambiguity in psychoanalytic writing. It is also argued that one particular reason for difficulties concerning the internal-external terminology is the existential anxiety awakened by the threat of the essential aloneness of man. The consciousness of this has a tendency to fade and lead to unclear terminology. The importance of the transitional world as a resting place from the hard reality of this essential aloneness is emphasized. The transitional world is also seen as a necessary part of psychoanalytical practice, as an aid in striving for the truth and reality, important goals of psychoanalysis. PMID- 20002818 TI - Postponing trauma: the dangers of telling. AB - Surviving a major historical trauma has consequences that are difficult to live with. Survivors who remain silent are often condemned to a desiccated existence, a dried-out life, a death in life. Survivors who speak out run an even greater risk. Telling their ghastly tale may trigger somatic consequences, psychotic episodes, or even suicide. As to the psychoanalytic cure, the free association it requires carries its own danger: negative therapeutic reaction in sometimes extreme forms. Avoidance of horror may turn into avoidance of life itself. Awful as it may seem, this avoidance of life may represent a victory over a menacing chaos. Should we as analysts accept the risk of endangering such a victory, no matter how unsatisfactory? The psychoanalytical injunction to speak out may trigger an upsurge of shame and terror. Is subjectivation always possible? This paper is about what happens when denial and splitting strategies are suspended, when 'crypts' are opened. Is there an analytic 'poros' allowing for a controlled return of affects? Is there a therapeutic solution to the problem of telling a wreckage without being caught in it? The dangers of 'telling' will be discussed in regard to new analytic strategies and new interpretive registers. When the 'silent psychic sharing' proves insufficient, some analysts go so far as to take part in the shame, share the grief, 'lend their own psyche', become a 'double' of the analysand, accept the existence of 'sanctuaries'. To what effect? PMID- 20002817 TI - Technique and final cause in psychoanalysis: four ways of looking at one moment. AB - This paper argues that if one considers just a single clinical moment there may be no principled way to choose among different approaches to psychoanalytic technique. One must in addition take into account what Aristotle called the final cause of psychoanalysis, which this paper argues is freedom. However, freedom is itself an open-ended concept with many aspects that need to be explored and developed from a psychoanalytic perspective. This paper considers one analytic moment from the perspectives of the techniques of Paul Gray, Hans Loewald, the contemporary Kleinians and Jacques Lacan. It argues that, if we are to evaluate these techniques, we must take into account the different conceptions of freedom they are trying to facilitate. PMID- 20002819 TI - When the third is dead: memory, mourning, and witnessing in the aftermath of the holocaust. AB - The origins of psychoanalysis, as well as the concerns of our daily endeavors, center on engagement with the fate of the unbearable - be it wish, affect, or experience. In this paper, I explore psychological states and dynamics faced by survivors of genocide and their children in their struggle to sustain life in the midst of unremitting deadliness. Toward this continuous effort, I re-examine Freud's theoretical formulations concerning memory and mourning, elaborate Andre Green's concept of the 'Dead Mother', and introduce more recent work on the concepts of the 'third' and 'thirdness'. Throughout, my thoughts are informed by our clinical experience with the essential role of witnessing in sustaining life after massive trauma. I bring aspects of all these forms of knowing to reflections about a poem by Primo Levi entitled Unfinished business and to our own never finished business of avoiding denial while living in an age of genocide and under the aura of uncontained destructiveness. PMID- 20002820 TI - Performative and enactive features of psychoanalytic witnessing: the transference as the scene of address. AB - This paper will attempt to broaden the conception of witnessing in analytic work with traumatized patients by extending the idea to incorporate the patient's developing and varied capacity for witnessing, as well as a witnessing that occurs within the analytic relationship itself. Actions occuring as part of traumatic repetition are understood to represent memory phenomena and are distinguised from dissociated self-state experience. These experiences are not therapeutically intended to be symbolized, but rather lived-through with the analyst, thus transforming the patient's own relation to the experience. I suggest that the scene in which this living-through takes place is the transference-countertransference matrix, and that it is the analytic encounter that allows traumatic repetition to take on the quality of a communication, an address to another, rather than remain meaningless reproduction. A clinical vignette illustrates the turning of trauma's imperative for witnessing into an address in the analytic encounter. PMID- 20002821 TI - Thoughts on representation in therapy of Holocaust survivors. AB - This paper presents the problems of representation and lack of representation in treating Holocaust survivors, through clinical vignettes and various theoreticians. The years of Nazi persecution and murder brought about a destruction of symbolization and turning inner and external reality into the Thing itself, the concrete, or, in Lacan's words, 'The Thing'. The paper presents two ideas related to praxis as well as theory in treating Holocaust survivors: the first is related to the therapist's treatment of the Holocaust nightmare expressing the traumatic events just as they happened 63 years previously; the second deals with the attempt at subjectification, in contrast to the objectification forced by the Nazis on their victims. PMID- 20002822 TI - Melanie Klein's letters addressed to Marcelle Spira (1955-1960). AB - Between 1955 and 1960, Melanie Klein wrote some 45 hitherto unpublished letters to Marcelle Spira, the Swiss psychoanalyst living at that time in Geneva. In 2006, after Spira's death, these letters were deposited with the Raymond de Saussure Psychoanalysis Centre in Geneva. They are the only known letters that Klein addressed to her psychoanalyst colleagues. Several topics are mentioned in them: (1) the meetings between the two women in Geneva and London; (2) Spira's contribution to Boulanger's translation into French of The Psychoanalysis of Children, which Klein herself carefully revised; (3) the papers that Klein was at that time working on, including Envy and Gratitude; (4) Spira's own work; (5) the difficulties that Spira, a Kleinian psychoanalyst who trained in Buenos Aires, was encountering in her attempt to be admitted to the Swiss Psychoanalytical Society; and (6) a few items of personal and family news. In addition to the invaluable historical information that these letters provide, they offer us a very moving epistolary self-portrait of Melanie Klein, enabling us to discover her personality in the final years of her life - she died in September 1960, just two months after writing her last letter to Spira. PMID- 20002823 TI - The French model of psychoanalytic training: Ethical conflicts. AB - Research on psychoanalytical education within the IPA may be clarified by reflecting on the ethic behind each of the three main models (Eitingonian, French and Uruguayan). In fact, the ethic underpinning psychoanalytical education, whatever the model, is confronted by irreducible conflicts between transmitting psychoanalysis by means of analytical experience or by means of academic teaching. Transmission by experience is essentially based on the ethic of psychoanalytic practice, which is difficult to regulate through institutional standards, whereas the academic aspect can be evaluated by objective and public criteria. The importance of both aspects and their relative weight in the training process depend on the conception of psychoanalysis underlying each model. This paper will look primarily at the French training model, the essentially analytical aspects of which favour the transmission of the very ethical foundations of psychoanalytic practice itself: the application of the method both as a working tool and as a tool of evaluation. It presupposes expanding the observation and analysis of transference beyond the framework of treatment to that of supervision. From this analysis, the paper will attempt to demonstrate how the French model proposes dealing with the inevitable conflicts between transmission by means of analysis and training by means of apprenticeship. PMID- 20002824 TI - On: On understanding projective identification in the treatment of psychotic states of mind. PMID- 20002826 TI - On: boys only! No mothers allowed. PMID- 20002828 TI - Between physical desire and emotional involvement: reflections on Frederic Fonteyne's film Une Liaison Pornographique. PMID- 20002835 TI - Calcium-binding proteins in skeletal muscles of the mdx mice: potential role in the pathogenesis of Duchenne muscular dystrophy. AB - Duchenne muscular dystrophy is one of the most common hereditary diseases. Abnormal ion handling renders dystrophic muscle fibers more susceptible to necrosis and a rise in intracellular calcium is an important initiating event in dystrophic muscle pathogenesis. In the mdx mice, muscles are affected with different intensities and some muscles are spared. We investigated the levels of the calcium-binding proteins calsequestrin and calmodulin in the non-spared axial (sternomastoid and diaphragm), limb (tibialis anterior and soleus), cardiac and in the spared extraocular muscles (EOM) of control and mdx mice. Immunoblotting analysis showed a significant increase of the proteins in the spared mdx EOM and a significant decrease in the most affected diaphragm. Both proteins were comparable to the cardiac muscle controls. In limb and sternomastoid muscles, calmodulin and calsequestrin were affected differently. These results suggest that differential levels of the calcium-handling proteins may be involved in the pathogenesis of myonecrosis in mdx muscles. Understanding the signaling mechanisms involving Ca(2+)-calmodulin activation and calsequestrin expression may be a valuable way to develop new therapeutic approaches to the dystrophinopaties. PMID- 20002836 TI - Increased calcium influx in the presence of ethanol in mouse pancreatic acinar cells. AB - The effects of alcohol on Ca(2+) signalling remains poorly understood. Here we have investigated the effects of acute ethanol exposure on Ca(2+) influx in mouse pancreatic acinar cells. Cells were loaded with fura-2 and the changes in fluorescence were monitored by spectrofluorimetry and imaging analysis. Stimulation of cells with 20 pM cholecystokinin evoked an oscillatory pattern in [Ca(2+)](c), both in the presence and in the absence of extracellular Ca(2+). Stimulation of cells with cholecystokinin in the presence of 50 mM ethanol led to a transformation of physiological oscillations into a single transient increase in [Ca(2+)](c). This effect was observed when Ca(2+) was present in the extracellular medium, and did not appear in its absence. Addition of 1 mM CaCl(2) to the extracellular medium, following release of Ca(2+) from intracellular stores by stimulation of cells with 1 nM cholecystokinin or 1 microM thapsigargin in the absence of extracellular Ca(2+), was followed by an increase in [Ca(2+)](c). Ca(2+) influx was increased in the presence of 50 mM ethanol. The anti-oxidant cinnamtannin B-1 (10 microM) or inhibition of alcohol dehydrogenase by 4-MP (1 mM), significantly reduced Ca(2+) influx evoked by cholecystokinin in the presence of ethanol. In summary, intoxicating concentrations of ethanol may lead to over stimulation of pancreatic acinar cells by cholecystokinin. This might be partially explained by the generation of reactive oxygen species and an increased Ca(2+) entry in the presence of ethanol. Potentially ethanol might lead to Ca(2+) overload, which is a common pathological precursor that is implicated in pancreatitis. PMID- 20002837 TI - A comparative study of the spatial distribution of mast cells and microvessels in the foetal, adult human thymus and thymoma. AB - Mast cells (MCs) are widely distributed in human and animal tissues and have been shown to play an important role in angiogenesis in normal and pathological conditions. Few data are available about the relationship between MCs and blood vessels in the normal human thymus, and there are virtually no data about their distribution and significance in thymoma. The aim of this study was to analyse the spatial distribution of MCs and microvessels in the normal foetal and adult thymus and thymoma. Twenty biopsy specimens of human thymus, including foetal and adult normal thymus and thymoma were analysed. Double staining with CD34 and mast cell tryptase was used to count both mast cells and microvessels in the same fields. Computer-assisted image analysis was performed to characterize the spatial distribution of MCs and blood vessels in selected specimens. Results demonstrated that MCs were localized exclusively to the medulla. Their number was significantly higher in thymoma specimens as compared with adult and foetal normal specimens respectively. In contrast the microvessel area was unchanged. The analysis of the spatial distribution and relationship between MCs and microvessels revealed that only in the thymoma specimens was there a significant spatial association between MCs and microvessels. Overall, these data suggest that MCs do not contribute significantly to the development of the vascular network in foetal and adult thymus, whereas in thymoma they show a close relationship to blood vessels. This could be an expression of their involvement not only in endothelial cells but also in tumour cell proliferation. PMID- 20002840 TI - Editorial comment to BK virus nephropathy: clinical experience in a university hospital in Japan. PMID- 20002838 TI - Myostatin and follistatin expression in skeletal muscles of rats with chronic heart failure. AB - Skeletal muscle abnormalities can contribute to decreased exercise capacity in heart failure. Although muscle atrophy is a common alteration in heart failure, the mechanisms responsible for muscle mass reduction are not clear. Myostatin, a member of TGF-beta family (transforming growth factor), regulates muscle growth and mass. Several studies have shown a negative correlation between myostatin expression and muscle mass. The aim of this study was to evaluate myostatin expression in skeletal muscles of rats with heart failure. As myostatin gene expression can be modulated by follistatin, we also evaluated its expression. Heart failure was induced by myocardial infarction (MI, n = 10); results were compared to Sham-operated group (n = 10). Ventricular function was assessed by echocardiogram. Gene expression was analyzed by real-time PCR and protein levels by Western blotting in the soleus and gastrocnemius muscles; fibre trophism was evaluated by morphometric analysis. MI group presented heart failure evidence such as pleural effusion and right ventricular hypertrophy. Left ventricular dilation and dysfunction were observed in MI group. In the soleus muscle, cross sectional area (P = 0.006) and follistatin protein levels (Sham 1.00 +/- 0.36; MI 0.18 +/- 0.06 arbitrary units; P = 0.03) were lower in MI and there was a trend for follistatin gene expression to be lower in MI group (P = 0.085). There was no change in myostatin expression between groups. In gastrocnemius, all MI group parameters were statistically similar to the Sham. In conclusion, our data show that during chronic heart failure, decreased skeletal muscle trophism is combined with unchanged myostatin and reduced follistatin expression. PMID- 20002841 TI - Primary seminoma of the prostate. AB - Abstract A 54-year-old gentleman was suspected of having sarcoma of the prostate because of his low serum prostate-specific antigen level (1.9 ng/mL) and an enlarged heterogeneous mass on computed tomography and magnetic resonance imaging scans. Pathological examination of the prostate needle biopsy indicated seminoma, which was confirmed with immunohistochemical staining. There was no evidence of disease in other areas on physical examination or on radiographic tests. Therefore, we diagnosed the case as a primary seminoma of the prostate, which was consequently treated with a total of three courses of bleomycin, etoposide and cisplatin chemotherapy. Complete response was obtained on computed tomography, magnetic resonance imaging and prostate needle re-biopsy. To our knowledge, there have only been five cases of primary seminoma of the prostate reported. PMID- 20002842 TI - A giant chromophobe renal cell carcinoma exceeding 10 kg. PMID- 20002843 TI - A case of juxtaglomerular cell tumor (reninoma) of the kidney treated with laparoscopic partial nephrectomy. PMID- 20002844 TI - A myelodysplastic male patient presenting with difficult catheterization as a result of a prostatic utricle cyst. PMID- 20002845 TI - Four-week fluconazole treatment is recommended for localized granulomatous cryptococcal prostatitis in patients with liver cirrhosis. PMID- 20002847 TI - Abstracts of the 4th UK Forum Stroke Conference. December 1-3, 2009. Glasgow, United Kingdom. PMID- 20002848 TI - Bronchoscopic evaluation of the mediastinum using endobronchial ultrasound: a description of the first 216 cases carried out at an Australian tertiary hospital. AB - BACKGROUND: Performance of linear probe endobronchial ultrasound-guided transbronchial needle aspiration (EBUS-TBNA) for staging non-small-cell lung cancer has been extensively studied. Alternate indications for its use are less well characterised, and performance in other clinical settings may differ. METHODS: We examined a prospectively collected cohort comprising the first 215 patients undergoing EBUS-TBNA at our institution. Patients were analysed according to the clinical and radiological indication for referral. We also examined the effect of the procedural learning curve on diagnostic sensitivity. RESULTS: A total of 215 patients underwent 216 EBUS-TBNA procedures. EBUS-TBNA returned adequate tissue for cytopathological analysis in 202 of 216 procedures (94%). Overall sensitivity for detection of malignancy was 0.92 (95% confidence interval 0.86-0.96); however, this varied according to the primary indication for EBUS-TBNA. Diagnostic sensitivity was high among all sub-groups, but the negative predictive value varied depending on the clinical indication for the procedure. We estimate 104 invasive surgical procedures and 32 inpatient admissions were avoided by use of EBUS-TBNA. Significant improvement in diagnostic performance was seen after 20 procedures were completed, and diagnostic accuracy did not peak until after 50 procedures. CONCLUSIONS: EBUS-TBNA is able to confirm accurately histologically a large number of disease processes, both malignant and benign, in all clinical indications studied. The procedure is safe even when carried out by proceduralists with minimal prior experience. Diagnostic performance continues to improve beyond 50 cases carried out. PMID- 20002849 TI - The current status of targeted therapy for non-small cell lung cancer. AB - Lung cancer accounts for more cancer-related deaths than any other malignancy in Australia and worldwide. Non-small cell lung cancer (NSCLC) accounts for about 85% of lung cancers and is associated with a 5-year survival of only 15%. Treatment with platinum-based doublets in the first-line setting and single agent chemotherapy in the second-line setting has improved survival and quality of life in patients with NSCLC. However, the benefits associated with chemotherapy are modest and serve to stress the need for novel therapeutic approaches. In the last decade a range of targeted therapies has been evaluated in NSCLC. Dramatic and often durable responses were seen in patients treated with the epidermal growth factor receptor (EGFR) tyrosine kinase inhibitors (TKI) gefitinib and erlotinib particularly in females, non-smokers, patients of East Asian ethnicity and those with adenocarcinomas - a group subsequently found to be enriched for tumours with activating EGFR mutations. Large randomized phase III trials have since established a role for EGFR TKI in the second- and third-line setting as well as a potential role for the monoclonal antibodies bevacizumab and cetuximab, directed at vascular endothelial growth factor and EGFR, respectively, in the combination with chemotherapy in the first-line setting. Recently it has been shown that patients with EGFR mutations may benefit from gefitinib in the first line setting. Other promising agents under evaluation are inhibitors of the insulin-like growth factor-1 receptor and inhibitors of recently described ALK gene rearrangements. PMID- 20002850 TI - Hospital doctors' attitudes towards older people. AB - BACKGROUND: Ageism among health professionals is increasingly recognized, but few studies investigated hospital doctors' attitudes towards older people. The aims of this study were to investigate hospital doctors' attitudes towards older people and to determine whether factors, which were identified in studies on other health professionals, influence hospital doctors' attitudes. METHODS: Hospital doctors who worked in General Medicine or Aged Care units in two tertiary public hospitals in metropolitan Victoria, Australia, in 2008 were surveyed with Fraboni's Scale of Ageism (FSA), a validated instrument used to investigate attitudes towards older people. Demographic data from participants were collected. RESULTS: Of the 235 questionnaires distributed, 122 were returned (overall response rate 51.9%). Response rate was highest among consultants (80.4%), followed by registrars (64.1%) and lowest among interns and residents (35.2%). The mean FSA score attained by the respondents was 61.5 (SD 11.0), representing a point between a neutral and a positive disposition. Doctors' characteristics that were associated with more positive attitudes towards older people included age of 30years or older (P < 0.001), female gender (P= 0.003), more senior in position (P < 0.001), postgraduate years of 10 or more (P < 0.001), previous working experience in Aged Care (P < 0.001), interest in Aged Care (P < 0.001) and more frequent social contacts with healthy older people (P < 0.001). CONCLUSION: Hospital doctors of different demographic features and background characteristics display different attitudes towards older people. These findings can be used to inform future development of undergraduate and postgraduate medical curricula and form a basis for future studies on the effectiveness of these interventions in improving doctors' attitudes. PMID- 20002851 TI - The Bosentan Patient Registry: long-term survival in pulmonary arterial hypertension. AB - BACKGROUND/AIMS: The Bosentan Patient Registry (BPR) was a prospective, multicentre, Australian registry funded by Actelion Pharmaceuticals. The primary aim of the registry was to collect survival data in patients with pulmonary arterial hypertension (PAH) treated with bosentan. METHODS: The BPR was initiated in 15 specialized PAH centres. All patients on or starting bosentan were invited to enrol. Treating physicians notified the registry if patients discontinued bosentan, because of either a change in therapy, transplantation, intervention or death. Survival data were validated against the Australian Institute of Health and Welfare National Death Index. RESULTS: Between 2004 and 2007, a total of 528 patients (mean age 59 +/- 17 years) were enrolled representing 69% of patients either previously taking or initiated on bosentan during that time. The BPR population was generally older with more advanced functional deficit than patients enrolled in randomized, placebo-controlled trials. Aetiology was idiopathic (iPAH) in 58% and connective tissue disease related (scleroderma (SSc) PAH) in 42%. For iPAH patients, World Health Organisation functional classes II, III and IV at enrolment was 8.2%, 66.4% and 20.5%, and for the SSc-PAH cohort, 3.2%, 75.8% and 17.9% respectively. The observed annual mortality was 11.8% in patients with iPAH and 16.6% in patients SSc-PAH. CONCLUSION: This large Australian registry provides 'real life' information on the characteristics and management of PAH in clinical practice. Treatment with bosentan improved survival outcomes in both iPAH and SSc-PAH compared with historical controls. Age, disease severity and aetiology were critical factors in determining clinical outcomes. PMID- 20002852 TI - Low prevalence of significant carotid artery disease on ultrasound in patients proceeding to coronary artery bypass surgery. AB - BACKGROUND: Cardiothoracic surgery is associated with an increased risk of perioperative stroke. Preoperative carotid ultrasonography can identify significant stenosis, but there is debate about the value of screening. The aims of this study were to (i) determine the prevalence of significant carotid artery disease in screened patients undergoing cardiothoracic surgery and (ii) correlate their ultrasonographic findings with perioperative strokes. METHODS: Retrospective analysis of 166 patients (118 men, 48 women) who underwent a preoperative carotid ultrasound and coronary artery bypass graft surgery (CABG) from 2004 to 2007. Perioperative strokes were recorded and compared with ultrasonographic and clinical data. A separate cohort of 1423 patients (1064 men, 359 women) who underwent CABG over the same period was also evaluated. RESULTS: Only 11 screened patients (6.6%) had significant (>70%) carotid artery disease and two of these underwent simultaneous carotid endarterectomy. There were five perioperative strokes in screened patients, four of which occurred in individuals with <50% disease. Compared with the non-screened cohort, ultrasound screened patients were older and more likely to have a prior stroke or transient ischaemic attack, hypertension, hypercholesterolaemia, peripheral vascular disease and/or renal impairment than non-screened patients. There was no significant difference in perioperative strokes compared with non-screened patients (3% vs 1.2% respectively, P= NS). CONCLUSION: There is a low prevalence of significant carotid artery disease in ultrasound screened patients. The risk of perioperative strokes in screened patients is low and not significantly different from non screened patients. PMID- 20002853 TI - Necrotizing fasciitis: review of 82 cases in South Auckland. AB - BACKGROUND: Early recognition of necrotizing fasciitis (NF) can be difficult, but is important as infections progress rapidly and have significant mortality. The aim of this study of patients with NF was to determine the clinical characteristics at presentation, causative pathogens and subsequent clinical outcome. METHODS: We retrospectively reviewed consecutive patients with NF presenting to Middlemore Hospital from January 2000 to June 2006. RESULTS: Eighty two patients were evaluated: 56% male, mean age 54.9 years (standard deviation 18.5), 40% Pacific Islanders. The site of infection was the lower limb in 46 (56%) patients, upper limb in 12 (15%) patients and perineum in 13 (16%) patients. Twenty-two (27%) patients were taking non-steroidal anti-inflammatory drugs, 29 (35%) had diabetes mellitus, 25 (30%) had gout and 17 (21%) had congestive heart failure. Forty-nine (60%) patients had a surgical procedure within 24 h of admission. Streptococcus pyogenes was isolated from tissue or blood cultures in 33 (40%) patients and 26 (32%) patients had polymicrobial infection. Twenty-five (30%) patients died, 17 (68%) within 72 h of admission. Independent predictors of mortality include congestive heart failure (P = 0.033) and a history of gout (P = 0.037). CONCLUSION: NF remains an important disease in our community with significant morbidity and mortality. Pacific Islanders were disproportionately represented. Early diagnosis of NF can be difficult and requires a high index of suspicion in all patients presenting with cellulitis or unexplained sepsis. Congestive heart failure and gout are independent predictors of mortality and patients with these conditions and sepsis need early assessment with more aggressive hospital triaging. PMID- 20002854 TI - Outpatient parenteral antimicrobial therapy-treated bone and joint infections in a tropical setting. AB - BACKGROUND: Osteoarticular infections are a primary indication for outpatient parenteral antimicrobial therapy (OPAT). The climate and geographical diversity of tropical Australia, together with the prevalence of melioidosis, disseminated gonococcal disease and community-acquired methicillin-resistant Staphylococcus aureus renders this a challenging environment in which to manage such infections. We evaluated patients managed by the Royal Darwin Hospital Hospital in the Home service for bone and joint infections. METHODS: A retrospective analysis of the therapeutic outcomes at the end of intravenous therapy was carried out for patients treated between 1 January 2006 and 15 September 2007. RESULTS: Fifty five patients were treated, including 21 (38%) indigenous Australians and 18 (33%) from remote communities. Baseline characteristics were similar to other published data, but there were two cases each of gonococcal septic arthritis and melioidosis. During treatment, 39 (71%) lived at home, with five (9%) of these receiving treatment at community clinics. Thirteen (24%) resided in self-care units in the hospital grounds. Three (5%) were managed at hostels or in prison. Median duration of parenteral therapy was 42 days, with a median of 22 days outside hospital, providing a total saving of 1307 bed-days. Clinical success at end of therapy was 84%, with no significant difference between indigenous and non indigenous cohorts. CONCLUSION: OPAT for osteoarticular infections is both feasible and effective in a tropical environment, including for indigenous patients. Extension of treatment to remote-dwelling patients is facilitated by the innovative use of self-care units and administration of treatment at remote clinics. PMID- 20002855 TI - CD38 as a prognostic marker in chronic lymphocytic leukaemia at a single New Zealand centre: patient survival in comparison to age- and sex-matched population data. AB - AIM: The aim of this study is to determine whether the analysis of CD38 expression by chronic lymphocytic leukaemia (CLL) cells provides useful additional prognostic information. METHODS: Clinical, laboratory, overall survival (OS) and treatment-free survival (TFS) data were collected on 130 CLL patients who had CD38 expression analysed at Canterbury Health Laboratories, New Zealand (NZ) during 1998-2008. RESULTS: The detection of any level of CD38 expression by CLL cells was associated with a significantly shorter OS and TFS. When analysis was restricted to Binet stage A patients, CD38 expression identified a subset of patients (21%) who, in common with Binet stage B/C patients, had a significantly shorter OS and TFS (P<0.0015), and a TFS at 4 years of <10%. In contrast, CD38-negative Binet stage A patients had an OS that was not significantly different from that of an age/sex-matched NZ population and a 5 year TFS of 77%. CONCLUSION: This study indicates that, when combined with clinical staging, the presence of any detectable CD38 expression can be used to further improve the identification of CLL patients with more aggressive disease (i.e. Binet stage B/C or Binet stage A and CD38 positive). This will allow better identification of those patients requiring more intensive monitoring and also allow improved patient counselling regarding prognosis. PMID- 20002856 TI - Study of compliance with a clinical pathway for suspected pulmonary embolism. AB - BACKGROUND/AIMS: Clinical pathways to guide the investigation of suspected pulmonary embolism have been increasingly adopted by emergency departments worldwide. This study evaluated the compliance with a clinical pathway that combines risk assessment (Wells score) with d-dimer, ventilation-perfusion scanning or computed tomographic pulmonary angiography (CTPA). The aims of this study were to identify factors that contribute to compliance and to assess patient outcomes and resource utilization. METHODS: Repeated retrospective chart reviews of 239 patients who underwent investigation for pulmonary embolism through our emergency department extracted patient demographics, pathway parameters and patient outcomes. A phone interview at 3-month follow up was carried out. RESULTS: Incidence of pulmonary embolism was 8.4% (n= 20). Compliance to the clinical pathway occurred in 120 subjects (50.2%). Non compliance occurred in 71 subjects (29.7%). Forty-eight subjects (20.1%) underwent risk assessments, but subsequent diagnostic tests did not conform to the stated pathway (partial compliance). Compliance was poor in subjects assessed by non-emergency department doctors (chi(2) = 27.95, P<= 0.001). Compliance occurred less in pregnant subjects (chi(2) = 7.27, P= 0.007) and those with chronic respiratory disease (chi(2) = 5.31, P= 0.021). Subjects in the compliant group were less likely to undergo CTPA (odds ratio 2.07 (1.16-3.70), P= 0.012). CONCLUSIONS: Compliance with this clinical pathway allowed emergency department doctors in an Australian university teaching hospital to complete diagnostic testing for suspected pulmonary embolism appropriately unless non-emergency department doctors became involved. Compliance with this pathway altered the distribution of diagnostic tests performed with less reliance on CTPA, but was not associated with better patient outcomes. PMID- 20002857 TI - Management of chronic obstructive pulmonary disease in Australia after the publication of national guidelines. AB - BACKGROUND: Information on the management of chronic obstructive pulmonary disease (COPD) in Australia, especially the extent of adherence to the COPD-X Plan, is sparse. AIM: To evaluate COPD patient adherence to treatment recommendations and healthcare provider adherence to the COPD-X Plan. METHODS: Cross-sectional study of patients admitted to a secondary care hospital with an acute exacerbation of COPD over a 6-month period. Data were collected from patient interviews and medical notes. RESULTS: Participants (n= 45) aged 72 +/- 11.5 years (mean +/- SD) had a mean FEV(1) % predicted 52.2 +/- 18.7. At the time 11 (24.4%) patients continued to smoke; 25 (55.6%) had never participated in a pulmonary rehabilitation programme; and 23 (51.1%) self-reported poor adherence to some COPD medications. Inhaler technique was deemed suboptimal in 25 (55.6%) patients. Only 11 (24.4%) patients had received any instructions from their doctor regarding management of exacerbations. The use of medications not supported by the COPD-X guidelines were: long-term prednisolone (11, 24.4%) and prophylactic antibiotics (3, 6.7%). CONCLUSION: Management of COPD in Australia by both patients and providers remains suboptimal despite the publication and wide dissemination of the COPD-X Plan, suggesting the need to intensify both patient and provider education in COPD management. PMID- 20002858 TI - Benchmarking opioids in the last 24 hours of life. AB - BACKGROUND/AIMS: Opioid dosages are titrated against symptoms with large variations between patients. For self-assessment of prescribing, and for clinical governance, it is important to know the normal prescribing patterns across New Zealand (NZ). The aims of this study were to document opioids and dosages given to patients in the last 24 h of life to identify normal practice and allow prescribers to reflect on their own practice. METHODS: A cross-sectional benchmarking design with retrospective chart review was carried out among 14 NZ hospices. Data (n = 352) on opioid dosages were analysed for inter-hospice variability (ANOVA). Parenteral morphine equivalent daily dose was used to analyse the dosage of different opioids. RESULTS: Overall, 95% of dying patients received an opioid. Of these 71% received morphine, 17% fentanyl, 10% methadone 9.5% oxycodone. The dosages delivered are conservative compared with international data, with a geometric mean of 47.8 mg. There was no significant difference in mean dosages of opioids prescribed between hospice teams. There was, however, a significant difference between the dosage of opioid for those on the Liverpool Care Pathway for the Care of the Dying and those who were not (63.1 44.4 mg), and between those with malignant and non-malignant disease (53.8-26.0 mg). Opioid footprints show different hospices have different patterns of opioid use. CONCLUSION: The parameters of what normal opioid prescribing is in the last days of life in NZ can be described from these data. There is value in repeating this exercise both for clinical governance and for professional reflection and self-assessment. PMID- 20002859 TI - Interacting effects of density and temperature on body size in multiple populations of Chinook salmon. AB - 1. The size individuals attain reflects complex interactions between food availability and quality, environmental conditions and ecological interactions. A statistical interaction between temperature and the density of conspecifics is expected to arise from various ecological dynamics, including bioenergetic constraints, if population density affects mean consumption rate or activity level. Density effects on behaviour or size-selective predation could also generate this pattern. This interaction plays an important role in bioenergetic models, in particular, and yet has not been documented in natural populations. 2. The lengths of 131 286 juvenile wild Chinook salmon (Oncorhynchus tshawytscha) across 13 populations spread throughout the Salmon River Basin, Idaho, USA over 15 years were compared to test whether juvenile density alters the relationship between body size and temperature. 3. Strong evidence for a negative interaction between mean summer temperature and density emerged, despite the relatively cool temperatures in this high elevation habitat. Growth correlated positively with temperature at lower densities, but the correlation was negative at the highest densities. 4. This is the first study to document this interaction at such a large spatial and temporal scale, and suggests that warmer temperatures might intensify some density-dependent processes. How climate change will affect individual growth rates in these populations will depend intimately on ecological conditions, particularly food availability and population dynamics. More broadly, the conditions that led to the interactions observed in our study - limited food availability and temperatures that ranged above those optimal for growth - likely exist for many other natural populations, and warrant broader exploration. PMID- 20002860 TI - Resource partitioning or reproductive isolation: the ecological role of body size differences among closely related species in sympatry. AB - 1. Body size differences among coexisting related species are common, but the actual effect of these differences in mitigating interspecific interactions, such as resource competition and reproductive interference, is poorly understood. 2. Local assemblages of the ground beetle genus Carabus (subgenus Ohomopterus) typically consist of two or more species of varying sizes. Through foraging and mating experiments using four Ohomopterus species in parapatry and sympatry, we examined whether interspecific body size differences are effective in partitioning food resources or reducing reproductive interference. 3. Because larval Ohomopterus feed exclusively on earthworms, body size differences may be related to partitioning earthworms of different sizes. However, larvae did not exhibit differences in selectivity or attack success on earthworms of different sizes based on larval body size, indicating little possibility of partitioning food by body size. 4. In contrast, interspecific mating behaviours, such as mate recognition, mounting, and copulation, were hindered when body size differences were large; copulation was frequently accomplished between parapatric species with smaller body size differences. 5. These results suggest that body size differences between species effectively reduce reproductive interference, rather than resource competition. Although body size differences in coexisting closely related species have been considered to function in resource partitioning, they may function primarily in reproductive isolation and thereby facilitate coexistence of species. PMID- 20002861 TI - Tits on the move: exploring the impact of environmental change on blue tit and great tit migration distance. AB - 1. In response to warmer spring conditions in Central Europe many migratory bird species have shifted their timing of breeding. Environmental change has also led to warmer winters, shortening the distance between the breeding grounds of migratory birds and their overwintering areas. 2. Here, we show that in response to warmer winters, blue tits (Cyanistes caeruleus), but not great tits (Parus major), breeding in Germany decreased their migration distance between 1964 and 1996. Understanding this difference provides insight into possible constraints and selection pressures involved in how species respond to environmental change. Here, we focus on their breeding ecology. 3. In a nest box population in southern Germany, both species laid their first clutch earlier with increasing spring temperature, but over the study period (1974-1999) blue tits showed a significant and stronger advancement in laying date than great tits. For both species, selection for earlier breeding did not vary with environmental change, indicating that early laying pairs did not do better than later laying pairs as spring temperature increased. 4. Blue tits in the nest box population were single brooded and existing hypotheses state that single-brooded species likely advance their laying date to match timing of reproduction with the advancing food peak in spring. We hypothesize that this might be one reason why blue tits adjusted their migration strategy as closer proximity to the breeding grounds in winter allows better prediction of the onset of spring. Ten per cent of great tits successfully produced second broods and their first clutch laying date is a compromise between first and second clutch laying date, which might be why great tits had not advanced their laying date nor altered their migration strategy. PMID- 20002862 TI - Analysis of variance with unbalanced data: an update for ecology & evolution. AB - 1. Factorial analysis of variance (anova) with unbalanced (non-orthogonal) data is a commonplace but controversial and poorly understood topic in applied statistics. 2. We explain that anova calculates the sum of squares for each term in the model formula sequentially (type I sums of squares) and show how anova tables of adjusted sums of squares are composite tables assembled from multiple sequential analyses. A different anova is performed for each explanatory variable or interaction so that each term is placed last in the model formula in turn and adjusted for the others. 3. The sum of squares for each term in the analysis can be calculated after adjusting only for the main effects of other explanatory variables (type II sums of squares) or, controversially, for both main effects and interactions (type III sums of squares). 4. We summarize the main recent developments and emphasize the shift away from the search for the 'right'anova table in favour of presenting one or more models that best suit the objectives of the analysis. PMID- 20002863 TI - Seasonal effects of Pacific-based climate on recruitment in a predator-limited large herbivore. AB - 1. Climate is an important factor influencing the population dynamics of large herbivores operating directly on individuals or through its effect on forage characteristics. However, the seasonal effect of climate may differ between forage- and predator-limited populations because of a climatic influence on predation rates. The influence of climate on predator-limited large herbivores is less well known than on forage-limited populations. Further, the effect of Pacific-based climate on large herbivore populations has been rarely assessed. 2. We investigated the effect of the Pacific Decadal Oscillation (PDO), across different seasons, on recruitment in 10 populations (herds) of mountain-dwelling caribou Rangifer tarandus caribou L. in the Yukon Territory, Canada. These low density populations occur in highly seasonal environments and are considered predator-limited with high neonatal calf mortality. Hence, in most years females do not spend resources through lactational support during the summer and resource intake is devoted to self-maintenance. We predicted that climate affecting environmental conditions at calving would have a strong effect on recruitment via its influence on predation rates. We also predicted that climatic conditions prior to conception could have an effect on recruitment through its influence on female fecundity. We modelled recruitment (n = 165) by seasonal PDO values using generalized linear mixed-effects models with herd-varying coefficients. 3. We found that recruitment variability was best explained by variation in winter climate (beta = 0.110, SE = 0.007) prior to birth (in utero) and May climate (beta = 0.013, SE = 0.006) at calving. There was little support for a pre conception climate effect influencing female body condition and hence fecundity. These results confirm that recruitment in these populations is limited by predation and that forage-limitation is not a significant factor in their population dynamics. There was considerable variability in herd-specific relationships between the PDO and recruitment. Incorporating herd-specific characteristics, such as variable predator densities or terrain characteristics within a herd range, may shed greater light on the complex relationship between climate and ungulate population dynamics. PMID- 20002864 TI - Soya bean tempe extracts show antibacterial activity against Bacillus cereus cells and spores. AB - AIMS: Tempe, a Rhizopus ssp.-fermented soya bean food product, was investigated for bacteriostatic and/or bactericidal effects against cells and spores of the food-borne pathogen Bacillus cereus. METHODS AND RESULTS: Tempe extract showed a high antibacterial activity against B. cereus ATCC 14579 based on optical density and viable count measurements. This growth inhibition was manifested by a 4 log CFU ml(-1) reduction, within the first 15 min of exposure. Tempe extracts also rapidly inactivated B. cereus spores upon germination. Viability and membrane permeability assessments using fluorescence probes showed rapid inactivation and permeabilization of the cytoplasmic membrane confirming the bactericidal mode of action. Cooked beans and Rhizopus grown on different media did not show antibacterial activity, indicating the unique association of the antibacterial activity with tempe. Subsequent characterization of the antibacterial activity revealed that heat treatment and protease addition nullified the bactericidal effect, indicating the proteinaceous nature of the bioactive compound. CONCLUSIONS: During fermentation of soya beans with Rhizopus, compounds are released with extensive antibacterial activity against B. cereus cells and spores. SIGNIFICANCE AND IMPACT OF STUDY: The results show the potential of producing natural antibacterial compounds that could be used as ingredients in food preservation and pathogen control. PMID- 20002865 TI - Exopolysaccharide analysis of biofilm-forming Candida albicans. AB - AIM: The major objective of the study was to analyse exopolysaccharide produced by a biofilm forming-clinical strain of Candida albicans. METHODS AND RESULTS: The biofilm-forming ability of C. albicans recovered from infected intrauterine devices (IUDs) was evaluated using XTT (2,3-bis[2-methoxy-4-nitro-5-sulfophenyl] 2H-tetrazolium-5-carboxanilide) reduction assay. The morphological characteristics of the biofilm were assessed using scanning electron microscopy (SEM), atomic force microscopy (AFM) and confocal laser scanning microscopy (CLSM). Biochemical characterization of the exopolysaccharide was carried out by gel permeation chromatography, gas chromatography (GC), Fourier transform infrared (FTIR) spectroscopy and nuclear magnetic resonance (NMR) spectroscopy. Microscopic studies of C. albicans biofilm revealed complex, heterogeneous three dimensional structure, in which yeast cells and hyphal elements were entrenched within exopolysaccharides matrix. Chromatographic analysis data indicated C. albicans exopolysaccharide (c. 300 kDa) to be made up of four major sugar units. The FTIR spectrum revealed specific absorbance of O-H, C-H, O=C=O, C=O, C-N and C C ring stretching. (1) H and (13) C NMR data showed the presence of beta (1->6) and beta (1->3) linkages in the exopolysaccharide chain that were assigned to alpha-D-glucose and beta-D-glucose, alpha-D-mannose, alpha-L-rhamnose and N acetyl glucosamine (beta-D-GlcNAc), respectively. CONCLUSIONS: Study suggested the production of a water soluble c. 300 kDa exopolysaccharide by C. albicans made up of glucose, mannose, rhamnose and N-acetyl glucosamine subunits. SIGNIFICANCE AND IMPACT OF THE STUDY: The study could assist in the development of novel therapeutics aimed at disrupting C. albicans biofilms that will translate into improved clearance of Candida-related infections. PMID- 20002866 TI - Physiological and molecular analysis of the stress response of Saccharomyces cerevisiae imposed by strong inorganic acid with implication to industrial fermentations. AB - AIMS: This work aimed to identify the molecular mechanism that allows yeast cells to survive at low pH environments such as those of bioethanol fermentation. METHODS AND RESULTS: The industrial strain JP1 cells grown at pH 2 was evaluated by microarray analysis showing that most of the genes induced at low pH were part of the general stress response (GSR). Further, an acid-tolerant yeast mutant was isolated by adaptive selection that was prone to grow at low pH in inorganic but weak organic acid. It showed higher viability under acid-temperature synergistic treatment. However, it was deficient in some physiological aspects that are associated with defects in protein kinase A (PKA) pathway. Microarray analysis showed the induction of genes involved in inhibition of RNA and protein synthesis. CONCLUSIONS: The results point out that low pH activates GSR, mainly heat shock response, that is important for long-term cell survival and suggest that a fine regulatory PKA-dependent mechanism that might affect cell cycle in order to acquire tolerance to acid environment. SIGNIFICANCE AND IMPACT OF THE STUDY: These findings might guide the construction of a high-fermentative stress tolerant industrial yeast strain that can be used in complex industrial fermentation processes. PMID- 20002867 TI - Unravelling the C3/C4 carbon metabolism in Ralstonia eutropha H16. AB - AIMS: Detailed knowledge about the enzymes responsible for conversion of C(3) and C(4) compounds will be helpful to establish the bacterial strain Ralstonia eutropha as platform for the production of biotechnologically interesting compounds. Although various studies about these enzymes were accomplished in the past, some contradicting information about the enzyme pattern in this bacterium still exists. To resolve these discrepancies, the C(3) /C(4) metabolism was reinvestigated after the genome sequence of this bacterium became available. METHODS AND RESULTS: In silico analysis of genome sequence revealed putative genes coding for NAD(P)(+) -dependent malic enzymes (Mae), phoshoenolpyruvate carboxykinase (Pck), phosphoenolpyruvate carboxylase (Ppc), phosphoenolpyruvate synthase (Pps) and pyruvate carboxylase (Pyc). Reverse transcription PCR revealed constitutive expression of mae and pck genes, whereas no transcripts of pyc and ppc were found. Expression of active NADP(+) -dependent MaeB and Pck and absence of Pyc and Ppc was confirmed by spectrophotometric enzyme assays. CONCLUSIONS: The data reported in this study suggest that two enzymes, (i) MaeB and (ii) Pck, mediate between the C(3) and C(4) intermediates in R. eutropha H16. The enzymatic conversion of pyruvate into phosphoenolpyruvate (PEP) is catalysed by Pps, and an NADH(+) -dependent Mdh mediates the reversible conversion of malate and oxaloacetate. SIGNIFICANCE AND IMPACT OF THE STUDY: An increased knowledge of the enzymes mediating between C(3) and C(4) intermediates in R. eutropha will facilitate metabolic engineering. PMID- 20002868 TI - Comparative susceptibility of planktonic and 3-day-old Salmonella Typhimurium biofilms to disinfectants. AB - AIMS: To compare the susceptibility of a 3-day-old biofilm and planktonic Salmonella to disinfectants at different exposure times. We hypothesize that Salmonella biofilms are more resilient to disinfectants compared to planktonic Salmonella. METHODS AND RESULTS: The susceptibility of planktonic cells to disinfectants was tested by a modified version of the Council of Europe suspension test EN 1276. Salmonella biofilms were formed using the Calgary Biofilm Device. Results show that 3-day-old Salmonella biofilms are less susceptible to the disinfectants benzalkonium chloride, chlorhexidine gluconate, citric acid, quaternary ammonium compounds, sodium hypochlorite (SH) and ethanol, compared to planktonic Salmonella. Surprisingly, the results also demonstrate that low concentrations of SH were more effective against a 3-day-old biofilm compared to high concentrations of SH. CONCLUSIONS: While all the disinfectants evaluated were able to reduce biofilm-associated cells at concentrations and contact times sufficient to eliminate planktonic cells, there were still sufficient viable cells remaining in the biofilm to cause further contamination and potential infection. SIGNIFICANCE AND IMPACT OF THE STUDY: Protocols for the use of chemical disinfectants need to include biofilm susceptibility testing. There is a requirement for an effective and standardized tool for determining the susceptibility of biofilms to disinfectants. PMID- 20002869 TI - Lipodystrophy - a sign for metabolic syndrome in patients of the HIV-HEART study. AB - BACKGROUND: After the introduction of antiretroviral therapy, the life expectancy of HIV patients has increased to more than 30 years after initial diagnosis. Cardiovascular disease now is an important cause of death in HIV-infected patients. PATIENTS AND METHODS: In the multicenter, prospective HIV-HEART study, 222 (38 %) patients suffered from lipodystrophy. Women were more often affected than men (41.5 % vs. 25.3 %). Patients with lipodystrophy were on average 5 years older and had been infected longer (10.4 vs. 6.6 years) then patients without lipodystrophy. RESULTS: Lipodystrophy in HIV patients was a clinical sign of cardiovascular risk factors like hyperlipidemia (total cholesterol 19 mg/dl higher, HDL 2.8 mg/dl lower, triglycerides 53 mg/dl higher) and type 2 diabetes (11.3 % vs. 2.8 %). Patients with lipodystrophy were more likely to be co infected with hepatitis B (34.7 % vs. 28.8 %, p = 0.122) or C (13.1 % vs. 9.3 %, p = 0.16) than patients without lipodystrophy. The quality of life was reduced in patients with lipodystrophy. In 6 of 8 scales of the SF-36 questionnaire, patients with lipodystrophy had lower scores. CONCLUSIONS: Lipodystrophy syndrome is an early warning system for a number of illnesses which reduce life expectancy. Dermatologists must help insure that HIV-infected patients receive treatment for these disorders. PMID- 20002870 TI - Molecular evidence that phylogenetically diverged ciliates are active in microbial mats of deep-sea cold-seep sediment. AB - Cold seeps are areas of the seafloor where hydrogen sulfide- and methane-rich fluid seepage occurs, often sustaining chemosynthetic ecosystems. It is well known that both archaea and bacteria oxidize sulfides and methane to produce chemical energy and that several endemic animals use this energy to thrive in cold seeps. On the other hand, there is little knowledge regarding diversity and ecology of microbial eukaryotes in this ecosystem. In this study we isolated environmental RNA and DNA from microbial mats of cold-seep sediment in Sagami Bay, Japan, and retrieved eukaryotic small-subunit ribosomal RNA sequences with polymerase chain reaction methods followed by clone library construction. Most RNA-derived clones obtained were from ciliates, although DNA-derived clones were mainly from the fungus Cryptococcus curvatus, suggesting that ciliates are active in the environment. The ciliate sequences were phylogenetically diverse, and represented eight known class lineages as well as undesignated lineages. Because most ciliates are bacterivorous, it is highly likely that the ciliates for which sequences were recovered play a role in the food web of this ecosystem as grazers of microbial mats. In addition, given that the environment studied is under highly reduced (anoxic) conditions, based on the prokaryotic community structure deduced from T-RFLP profiles, the ciliates detected may be obligatory or facultative anaerobes. PMID- 20002871 TI - In vitro cultivation and morphological characterization of phloemic trypanosomatids isolated from coconut trees. AB - Plant trypanosomatids cause lethal vascular wilting in palms of the Arecaceae family. Infections, affecting plants in South and Central America, can result in significant economic loss. The study of trypanosomatids that cause these diseases has been complicated due to the inability to culture these organisms for in vitro analyses. To develop a protocol that would facilitate studies of trypanosomatids, continuous in vitro cultures of phloemic trypanosomatids were established from apical stems of diseased coconut trees collected in endemic and non-endemic regions of Brazil (the states of Bahia and Rio de Janeiro, respectively). Although attempts at establishing axenic cultures were unsuccessful, it was found that trypanosomatid co-cultures could be successfully established and maintained. The procedure was to preculture media with 10(4)Aedes albopictus cells in Grace's medium supplemented with 10% heat-inactivated fetal bovine serum (without antibiotics or fungicides) for 3 d before adding 10(6) trypanosomatids/ml harvested from either fresh apical stem extracts or with 2 mm(3) fragments of coconut apical stems. By day 7 under these conditions the parasites grew exponentially. Using this strategy, two isolates were identified and have been maintained in our laboratory for over 400 passages, demonstrating the efficacy of this culturing procedure. In situ the organisms were observed in vascular bundles and inside sieve elements of the phloem of diseased palms. In vitro parasites retained their mobility. Morphometric analysis revealed differences between Bahia and Rio de Janeiro isolates. PMID- 20002872 TI - Neuroendocrine cells and associated malignancies of the oral mucosa: a review. AB - Neuroendocrine cells of the oral mucosa constitute an under-recognized component of the diffuse neuroendocrine system with diverse subpopulations and elusive biologic roles in the oral cavity. Primary malignant oral tumors that show a neuroendocrine phenotype display histomorphologic heterogeneity thereby giving rise to a spectrum of lesions in this rare category of oral malignancy. These lesions can be divided into neuroendocrine carcinomas (NECs) of small cell or non small cell type. The former is further subdivided into the Merkel cell type or the pulmonary type while the latter includes atypical carcinoid tumor and large cell NEC. All histologic subtypes of oral NEC appear to have a strong predilection for men in their fifth or sixth decade and arise predominantly in the non-keratinized oral mucosa. The biologic behavior of oral Merkel cell carcinomas appears to be more aggressive than those of skin. It remains to be determined whether histologic categorization of the remaining tumor subtypes is predictive of patient survival in oral neuroendocrine tumors. PMID- 20002873 TI - NFATc1 and TNFalpha expression in giant cell lesions of the jaws. AB - BACKGROUND: Activation mutations of SH3BP2 gene have been demonstrated in cherubism and central giant cell lesion (CGCL). In the present study we first attempted to investigate the SH3BP2 gene in peripheral giant cell lesion (PGCL). The effect of SH3BP2 gene mutations on the transcription of the downstream genes nuclear factor of activated T cells (NFATc1) and the cytokine tumor necrosis factor-alpha (TNF-alpha) was also investigated together with the immunolocalization of NFATc1 protein in a set of cases of PGCL, CGCL and cherubism with and without SH3BP2 mutation. METHOD: Fresh samples of five PGCL, five CGCL and one cherubism cases were included in this study. One of the samples of CGCL presented a somatic heterozygous mutation c.1442A>T in exon 11. The cherubism case showed a heterozygotic substitution c.320C>T in both blood and lesion. These mutations were previously published. All coding and flanking regions of the SH3BP2 gene were sequenced in the cases of PGCL. The real-time polymerase chain reaction (RT-PCR) was performed to analyze the transcription of NFATc1 and TNF-alpha genes. The immunohistochemical analysis of the NFATc1 protein was also performed. RESULTS: No SH3BP2 gene mutation was found in PGCL. The RT-PCR showed increased expression of NFATc1 and decreased transcription of TNF-alpha in all the samples. The immunohistochemical analysis of the NFATc1 protein showed a predominant nuclear staining in the multinucleated giant cells. CONCLUSION: The development of giant cells lesions of the jaws and cherubism are possibly mediated by overexpression of NFAT in the nucleus of the multinucleated cells. PMID- 20002874 TI - Association between oral health-related and general health-related quality of life in subjects attending dental offices in Germany. AB - OBJECTIVES: To evaluate the GHRQoL and OHRQoL of patients attending dental offices in Germany and to determine correlation coefficients between SF (Short Form)-12 and OHIP (Oral Health Impact Profile)-14 scores. METHODS: A total of 10,342 dental offices were randomly selected. Each of the 1,113 that consented to participate received 20 questionnaires to be filled in by a convenience sample of the patients. The questionnaire included the OHIP-14-form for OHRQoL as well as the SF-12-form for GHRQoL. RESULTS: A total of 12,392 completed questionnaires were analyzed. The mean age of the participants (64.9 percent female, 35.1 percent male) was 44.25 years. The mean summary score of OHIP-14 was 6.30 (SD 7.46). The mean physical component summary scale (PCS) of the SF-12 was 51.15 (SD 7.23) and the mental component summary scale (MCS) was 50.17 (SD 8.55). The variance of PCS and MCS could be explained to 10 percent each by oral health related quality of life (r2 = 0.095 and 0.101, P < 0.001). CONCLUSION: OHRQoL is considerably related to GHRQoL. PMID- 20002875 TI - The dental office visit as a potential opportunity for diabetes screening: an analysis using NHANES 2003-2004 data. AB - OBJECTIVES: The bidirectional relationship between periodontitis and diabetes suggests that the dental visit may offer a largely untapped opportunity to screen for undiagnosed diabetes. To better examine this potential opportunity, data from the National Health and Nutrition Examination Survey (NHANES) 2003-2004 were used to determine if a larger proportion of patients with periodontal disease as compared with those without periodontitis would be recommended for screening according to American Diabetes Association (ADA) guidelines. The data were also used to determine whether at-risk individuals with periodontitis visited a dental professional recently, so that they could avail themselves of this opportunity for screening, if offered. METHODS: Data to perform these analyses were collected from 2,923 subjects aged 20 and older who reported that they were never told that they had diabetes, had a periodontal examination, and had sufficient data to compute body mass index. Descriptive statistics, t-tests, and chi-square analyses that compared those with and without periodontitis were extrapolated to the US population. RESULTS: A total of 62.9 percent of those without periodontitis and 93.4 percent of those with periodontal disease met ADA guidelines for diabetes screening. Of those at-risk with periodontal disease, 33.9 percent had seen a dentist in the past 6 months, 50 percent in the past year, and 60.4 percent in the past 2 years. CONCLUSIONS: As almost all individuals with periodontitis would have been recommended for diabetes screening, and many at-risk persons with periodontal disease recently visited a dentist, our data suggest that the dental visit provides an important potential venue for this screening. PMID- 20002876 TI - Dental care expenditures and retirement. AB - OBJECTIVES: To examine the relationship of dental care coverage, retirement, and out-of-pocket (OOP) dental expenditures in an aging population, using data from the Health and Retirement Study (HRS). METHODS: We estimate OOP dental expenditures among individuals who have dental utilization as a function of dental care coverage status, retirement, and individual and household characteristics. We also estimate a multivariate model controlling for potentially confounding variables. RESULTS: Overall, mean OOP dental expenditures among those with any spending were substantially larger for those without coverage than for those with coverage. However, controlling for coverage shows that there is little difference in spending by retirement status. CONCLUSIONS: Although having dental coverage is a key determinant of the level of OOP expenditures on dental care; spending is higher among those without coverage than those who have dental insurance. We also found that while retirement has no independent effect on OOP dental expenditures once controlling for coverage, dental coverage rates are much lower among retirees. PMID- 20002877 TI - Factors associated with the recruitment and retention of dentists in the public sector. AB - OBJECTIVES: There is an increasing demand for public dental services in Australia, with many community dental clinics unable to meet this demand because of an inadequate number of dentists in the workforce. The aim of this study was to identify factors contributing to the recruitment and retention of dentists in the public sector. METHODS: A postal questionnaire survey of 180 dentists (response rate 75.6 percent) working in the Victorian public sector was undertaken to investigate the characteristics of public sector dentists, job satisfaction, remuneration, perceptions of public dentistry, future career intentions, and issues that relate to recruitment and retention of staff. RESULTS: Victorian public dentists' main reason for entering the public sector was to work in a community-based setting in a supportive and mentored environment. The main factors related to dentists leaving the public sector were poor remuneration, lack of clinical experience, and frustration with administrative policies. Victoria's oral health workforce shortages in the public sector are mainly attributed to retention issues. The potential for mentoring and a desire for helping those in need were factors attracting dentists to work in the public sector. CONCLUSION: There was a disproportionate number of female dentists in the public sector compared with the general population, and female dentists had a lower mean salary than male dentists regardless of experience. A range of factors were associated with retention, and gradual frustration because of poor remuneration and lack of professional autonomy were significant reasons for the decision to leave the public sector. PMID- 20002878 TI - Association between obesity and dental caries in a group of preschool children in Mexico. AB - OBJECTIVE: The aim of this study was to determine the association between obesity and caries by utilizing the data of a cohort of preschool children aged 4-5 years. METHODS: Data were obtained from a cohort of 1,160 children. Dental caries detection was performed according to the World Health Organization criteria. The caries index was measured as the number of decayed (d), extracted (e), and filled (f) teeth (t) (deft), or surfaces (defs). The body mass index (BMI) in units of kg/m2 was determined, and children were categorized according to age- and gender specific criteria as normal weight (5th-85th percentile), at-risk overweight (> or = 85th-<95th percentile), and overweight (> or = 95th percentile). Odds ratios were determined for at-risk overweight and overweight children using logistic regression. RESULTS: The prevalence of dental caries was 17.9 percent. A slightly higher percentage of dental caries was found in boys (19.6 percent) than in girls (16.4 percent). From the total sample, the mean BMI was 17.10 +/- 3.83. Approximately 53.7 percent of children were classified as normal weight, 14.2 percent as at-risk overweight, and 32.1 percent as overweight. At-risk overweight children were higher among girls (17.1 percent) than among boys (11.3 percent). When adjusted for covariates, the logistic regression model showed that there was a significant association between at-risk overweight children (P < 0.001), overweight children (P < 0.001), and caries in the primary dentition. Mean (SD) deft value of the sample was 1.08 (2.34), while the corresponding defs value was 1.43 (3.29). CONCLUSION: Obesity appears to be associated with dental caries in the primary dentition of preschool Mexican children. PMID- 20002879 TI - Transport-dependent endocytosis and turnover of a uric acid-xanthine permease. AB - In this work we unmask a novel downregulation mechanism of the uric acid/xanthine transporter UapA, the prototype member of the ubiquitous Nucleobase-Ascorbate Transporter family, directly related to its function. In the presence of substrates, UapA is endocytosed, sorted into the multivesicular body pathway and degraded in vacuoles. Substrate-induced endocytosis, unlike ammonium-induced turnover, is absolutely dependent on UapA activity and several lines of evidence showed that the signal for increased endocytosis is the actual translocation of substrates through the UapA protein. The use of several UapA functional mutants with altered kinetics and specificity has further shown that transport-dependent UapA endocytosis occurs through a mechanism, which senses subtle conformational changes associated with the transport cycle. We also show that distinct mechanisms of UapA endocytosis necessitate ubiquitination of a single Lys residue (K572) by HulA(Rsp5). Finally, we demonstrate that in the presence of substrates, non-functional UapA versions can be endocytosed in trans if expressed in the simultaneous presence of active UapA versions, even if the latter cannot be endocytosed themselves. PMID- 20002880 TI - Screening examination and treatment of Trichophyton tonsurans infection in judo athletes affiliated with the University Judo Federation of Tokyo. AB - In Japan, Trichophyton tonsurans infection has become an increasing problem among combat sports participants. We investigated the prevalence of T. tonsurans infection in athletes affiliated to judo clubs in the 21 First Division universities that were registered with the University Judo Federation of Tokyo in 2008. Study procedures performed by the subjects included (i) completion of a questionnaire concerning lifestyle, risk factors for tinea corporis and medical history; (ii) scrubbing the scalp with a circular hairbrush to obtain samples for fungal culture; (iii) anti-fungal treatment as recommended by a dermatologist, based on the number of fungal colonies isolated from the hairbrush; and (iv) repeat testing using the hairbrush method 3 months after treatment recommendations were received. Of 902 study subjects, 102 (11.3%) yielded positive hairbrush culture results. Of these, 14 individuals (13.7%) had tinea corporis; the remainder were asymptomatic. Conversion to negative fungal culture was observed in 85 of 96 culture-positive individuals who performed the second hairbrush culture test following treatment. Control of T. tonsurans infection among judo athletes could be achieved by educating athletes, trainers and coaches in judo clubs concerning detection, prevention, and treatment of T. tonsurans infection. PMID- 20002881 TI - Prevalence of Candida dubliniensis among cancer patients in Kuwait: a 5-year retrospective study. AB - Despite close genetic and phenotypic relationship of Candida dubliniensis with Candida albicans, its role in human disease is mostly restricted to oral colonisation, particularly among HIV-infected patients. The prevalence of C. dubliniensis in association with other disease conditions has been infrequently reported. In this study, we present data on the prevalence of C. dubliniensis among yeast species isolated from cancer patients over a 5-year period. A total of 1445 yeast isolates recovered from respiratory specimens, blood, urine and oral swabs were analysed. Candida dubliniensis isolates were provisionally identified by phenotypic methods and their identity was further confirmed by species-specific amplification and/or sequencing of internally transcribed spacer region of rDNA. Antifungal susceptibility for fluconazole was determined by Etest. The number of isolates identified as C. dubliniensis, C. albicans and other yeast species were 71 (4.9%), 862 (59.6%) and 512 (35%) respectively. All the C. dubliniensis isolates originated from respiratory (5.9%) or oral (3.2%) specimens with an overall prevalence of 4.9%, and were found to be susceptible to fluconazole. The isolation of C. dubliniensis from respiratory or oral specimens and not from blood or urine specimens suggests that this species has preference to colonise these sites of human body. PMID- 20002882 TI - Molecular analysis of the cutaneous Malassezia microbiota from the skin of patients with atopic dermatitis of different severities. AB - Cutaneous Malassezia is an exacerbating factor in patients with atopic dermatitis. We analysed the Malassezia microbiota of adult patients with head and neck atopic dermatitis of different severities (mild, moderate and severe). Of the nine human-associated Malassezia species, the number detected was similar (3.5-4.2 species per case) among the members of all severity groups. However, the ratio of the two major Malassezia species, M. globosa and M. restricta, was different in the severe group. PMID- 20002883 TI - Clinically relevant drug interactions of current antifungal agents. AB - Antifungal agents are often prescribed in critically ill patients who are receiving many other medications. When using systemic antifungals, clinicians may possess susceptibility data and they are typically aware of the potential toxicity of these agents. However, the myriad of potential drugs that antifungal agents can interact with is daunting and can be confusing. This article reviews the pharmacokinetic properties of antifungal agents and their clinically relevant drug interactions. The antifungal agents differ markedly in their pharmacokinetic properties and in how they interact with other medicines. The amphotericin B formulations interact with other medicines primarily by reducing their renal elimination or producing additive toxicities. The azoles interact with other medicines primarily by inhibiting biotransformation or by affecting drug distribution and elimination. The echinocandins have the lowest propensity to interact with other medicines. The clinical relevance of antifungal-drug interactions varies substantially. While certain interactions are benign and result in little or no untoward clinical outcomes, others can produce significant toxicity or compromise efficacy if not properly managed through monitoring and dosage adjustment. However, certain interactions produce significant toxicity or compromise efficacy to such an extent that they cannot be managed and the particular combination of antifungal and interacting medicine should be avoided. PMID- 20002884 TI - Tinea capitis: still an unsolved problem? AB - Tinea capitis is a fungal infection specifically involving the scalp and hair. It is the most common dermatophyte infection in children under 12 years of age, with a predominance in those of sub-Saharan African descent. Common signs include hair loss, scaling, erythema and impetigo-like plaques. Adults may also be affected, but to a lesser degree. The causative species are from the Microsporum and Trichophyton genera. Limited treatment options and diverse modes of transmission complicate the clinician's ability to address this disease adequately. Although dermatophytes are ubiquitous in our environment and tinea capitis is common, therapeutic options can be utilised to reduce morbidity. PMID- 20002885 TI - The 2009 stock conference report: inflammation, obesity and metabolic disease. AB - Obesity is linked with many deleterious health consequences and is associated with increased risk of chronic disease including type 2 diabetes, atherosclerosis and certain forms of cancer. Recent work has highlighted the impact of obesity to activate inflammatory gene networks and suggests a causal function of inflammation in the pathogenesis of the metabolic syndrome. Since 2005, when Dr Gokhan Hotamisligil chaired the fourth Stock Conference in Istanbul, Turkey, entitled 'Obesity and Inflammation', there has been an explosion of studies investigating the relationship between obesity, inflammation and substrate metabolism. The exuberance surrounding this field of research is exemplified by the body of work that has been published in these past 4 years, including over 1400 publications. During this time, several novel mechanisms relating to cellular inflammation have been uncovered including the role of the hematopoietic system, toll-like receptor activation, endoplasmic reticulum stress and very recently T-cell activation in obesity-induced insulin resistance. These discoveries have led us to rethink cellular nutrient sensing and its role in inflammation and metabolic disease. Despite burgeoning investigation in this field, there still remain a number of unanswered questions. This review that evolved from the 2009 Stock Conference summarizes current research and identifies the deficiencies in our understanding of this topic. The overall goal of this Stock Conference was to bring together leading investigators in the field of inflammation and obesity research in the hope of fostering new ideas, thus advancing the pursuit of novel therapeutic strategies to reduce disease risk and or better treat chronic disease including type 2 diabetes, cardiovascular disease and cancer. PMID- 20002886 TI - Survival of transvenous ICD leads in young patients. AB - BACKGROUND: In adults, transvenous implantable cardioverter defibrillator (ICD) lead failure rates are significant, and their occurrence increases with time from implant. There are limited data in children. The goal of this study was to assess lead survival in young patients undergoing ICD implantation at a single center. METHODS: Records of patients under 21 years old with transvenous ICD leads implanted at our center from June 1997 to August 2007 were retrospectively reviewed. Age, weight, height, diagnosis, lead and generator model, venous access technique, generator position, pacing thresholds, lead impedance, and R wave size were recorded. "Lead failure" was defined as any lead problem requiring surgical intervention to restore proper function to the ICD system. RESULTS: Seventy-one transvenous leads were included (70 patients). Average age at implant was 14.8 years (range 5.7-19.5). All the devices were implanted by a single operator (HMS). Venous access was obtained via cephalic cutdown in 66/71. Mean follow-up time was 2.8 years (range 0.2-7.8 years, median 2.3 years). There were no infections requiring explantation. There were four lead failures. Three were lead fractures, occurring 12, 13, and 19 months after implant. The fourth lead failed when an arrhythmia was not appropriately detected, and a second dedicated rate sensing lead was thus implanted. Univariate analysis did not identify any variable to be a significant predictor of lead failure. Kaplan-Meier survival analysis demonstrated 5-year lead survival at 89.6%. CONCLUSIONS: ICD lead survival in children, when performed by an experienced operator, is similar to that found in adults. PMID- 20002887 TI - Inducible expression of (V600E) Braf using tyrosinase-driven Cre recombinase results in embryonic lethality. AB - We recently demonstrated that expression of (V600E)Braf in mature mouse melanocytes induces melanoma. Here, we show that expression of (V600E)Braf using the tyrosinase promoter leads to an unexpected embryonic lethality, with the animals dying before, at, or shortly after birth. The mice suffer from a range of developmental defects in the skin, the brain, the eyes and the heart, tissues that are normally colonized by melanocytes. We show that the (V600E)Braf expressing cells are potential melanocytic precursors that are fully transformed, suggesting that (V600E)Braf stimulates proliferation and blocks differentiation of these cells. Our data suggests that the presence of these cells in the organs that are normally occupied by melanocytes leads to severe developmental disruption, resulting in catastrophic defects and leading to death of the individual. PMID- 20002888 TI - The PGF(2alpha) receptor FP is lost in nevi and melanoma. PMID- 20002889 TI - A delphi method expert survey to derive standards for flood damage data collection. AB - For the purpose of flood damage analyses reliable, comparable, comprehensive, consistent, and up-to-date data are an indispensable need. Like in many other countries a database with this kind of datasets does not exist in Germany. To establish it, standards have to be set for flood damage data collection. We approached this problem by questioning experts about their information needs for flood damage analysis. This survey is done by applying a Delphi survey approach. The aptitude of the Delphi approach to assess, structure, and standardize expert knowledge is evaluated in this article. In the survey a panel of 55 experts working in the field of flood damage analysis for insurances, engineering companies/consultancy, public water management, and universities and other scientific institutions helped to identify common information needs. The multi step Delphi method proved to reduce the deviation of answers thereby enabling consensual results and also enhanced the quality by modifying group answers in the direction of experience based answers. There was also a high level of congruence in information needs between experts from different fields of employment that allowed the derivation of common standards. PMID- 20002890 TI - Quantitative risk assessment relating to adventitious presence of allergens in food: a probabilistic model applied to peanut in chocolate. AB - Peanut allergy is a public health concern, owing to the high prevalence in France and the severity of the reactions. Despite peanut-containing product avoidance diets, a risk may exist due to the adventitious presence of peanut allergens in a wide range of food products. Peanut is not mentioned in their ingredients list, but precautionary labeling is often present. A method of quantifying the risk of allergic reactions following the consumption of such products is developed, taking the example of peanut in chocolate tablets. The occurrence of adventitious peanut proteins in chocolate and the dose-response relationship are estimated with a Bayesian approach using available published data. The consumption pattern is described by the French individual consumption survey INCA2. Risk simulations are performed using second-order Monte Carlo simulations, which separately propagates variability and uncertainty of the model input variables. Peanut allergens occur in approximately 36% of the chocolates, leading to a mean exposure level of 0.2 mg of peanut proteins per eating occasion. The estimated risk of reaction averages 0.57% per eating occasion for peanut-allergic adults. The 95% values of the risk stand between 0 and 3.61%, which illustrates the risk variability. The uncertainty, represented by the 95% credible intervals, is concentrated around these risk estimates. Children have similar results. The conclusion is that adventitious peanut allergens induce a risk of reaction for a part of the French peanut-allergic population. The method developed can be generalized to assess the risk due to the consumption of every foodstuff potentially contaminated by allergens. PMID- 20002891 TI - A risk assessment model to evaluate the role of fecal contamination in recreational water on the incidence of cryptosporidiosis at the community level in Ontario. AB - A quantitative microbial risk assessment model was developed to simulate the role of recreational water contact in the transmission of cryptosporidiosis in a model Ontario community. Stochastic simulations were based on plausible modes of contamination of a pool (literature derived), river (site-specific), and recreational lakes (literature derived). The highest estimated risks of infection were derived from the (highly contaminated) recreational lake scenario, considered the upper end for risk of infection for both children (10 infections per 1,000 swims [5 per thousand: two infections per 1,000 swims; 95 per thousand: three infections per 100 swims]) and adults (four infections per 1,000 swims [5 per thousand: four infections per 1,000 swims; 95 per thousand: one infection per 100 swims]). Simulating the likely Cryptosporidium oocyst concentration in a lane pool that a child would be exposed to following a diarrheal fecal release event resulted in the third highest mean risk of infection (four infections per 10,000 swims [5 per thousand: three infections per 100,000; 95 per thousand: 10 infections per 10,000 swims]). The findings from this study illustrate the need for systematic and standardized research to quantify Cryptosporidium oocyst levels in Canadian public pools and recreational beaches. There is also a need to capture the swimming practices of the Canadian public, including most common forms and frequency measures. The study findings suggest that swimming in natural swim environments and in pools following a recent fecal contamination event pose significant public health risks. When considering these risks relative to other modes of cryptosporidiosis transmission, they are significant. PMID- 20002892 TI - Consumer confidence in the safety of food and newspaper coverage of food safety issues: a longitudinal perspective. AB - This study develops a longitudinal perspective on consumer confidence in the safety of food to explore if, how, and why consumer confidence changes over time. In the first study, a theory-based monitoring instrument for consumer confidence in the safety of food was developed and validated. The monitoring instrument assesses consumer confidence together with its determinants. Model and measurement invariance were validated rigorously before developments in consumer confidence in the safety of food and its determinants were investigated over time. The results from the longitudinal analysis show that across four waves of annual data collection (2003-2006), the framework was stable and that the relative importance of the determinants of confidence was, generally, constant over time. Some changes were observed regarding the mean ratings on the latent constructs. The second study explored how newspaper coverage of food safety related issues affects consumer confidence in the safety of food through subjective consumer recall of food safety incidents. The results show that the newspaper coverage on food safety issues is positively associated with consumer recall of food safety incidents, both in terms of intensity and recency of media coverage. PMID- 20002893 TI - Intelligent adversary risk analysis: a bioterrorism risk management model. AB - The tragic events of 9/11 and the concerns about the potential for a terrorist or hostile state attack with weapons of mass destruction have led to an increased emphasis on risk analysis for homeland security. Uncertain hazards (natural and engineering) have been successfully analyzed using probabilistic risk analysis (PRA). Unlike uncertain hazards, terrorists and hostile states are intelligent adversaries who can observe our vulnerabilities and dynamically adapt their plans and actions to achieve their objectives. This article compares uncertain hazard risk analysis with intelligent adversary risk analysis, describes the intelligent adversary risk analysis challenges, and presents a probabilistic defender attacker-defender model to evaluate the baseline risk and the potential risk reduction provided by defender investments. The model includes defender decisions prior to an attack; attacker decisions during the attack; defender actions after an attack; and the uncertainties of attack implementation, detection, and consequences. The risk management model is demonstrated with an illustrative bioterrorism problem with notional data. PMID- 20002894 TI - Management of blood supplies during an influenza pandemic. AB - BACKGROUND: Blood supplies are delicate resources, particularly vulnerable to incidents affecting the health of donors. The critical impact of a pandemic on the availability of red blood cells (RBCs) has been demonstrated in previous research; however, a detailed estimate of the expected deficit is missing. This has become a priority issue in the face of the current influenza pandemic. STUDY DESIGN AND METHODS: Data from several major blood donation services were used to analyze management of blood supplies in Germany. Routine management of RBCs was extrapolated to epidemic and pandemic situations using computer simulations with a mathematical model that allows for analysis of deficits in blood supplies. RESULTS: Routine management and distribution of RBCs are driven by supply, which has marked fluctuations but does not appear to have seasonality. There seems to be a remarkable elasticity in the demand for RBCs that helps to mitigate minor crises in supply, but this is likely to be overstretched during a severe pandemic. CONCLUSION: The supply-driven management of RBCs in Germany implies that assessment of severity of shortages due to a pandemic depends on detailed knowledge about the fraction of transfusions that do not allow for postponement. Pandemic preparedness should include criteria for prioritization of transfusions. PMID- 20002895 TI - Molecular dissection of abnormal wound healing processes resulting in keloid disease. AB - Keloids are locally aggressive scars that typically invade into healthy surrounding skin and cause both physical and psychosocial distress to the patient. These pathological scars occur following minimal skin trauma after a variety of causes including burns and trauma. Although the pathogenesis of keloid disease is not well understood, it is considered to be the end product of an abnormal healing process. The aim of this review was to investigate the molecular and cellular pathobiology of keloid disease in relation to the normal wound healing process. The molecular aberrances in keloids that correlate with the molecular mechanisms in normal wound healing can be categorized into three groups: (1) extracellular matrix proteins and their degradation, (2) cytokines and growth factors, and (3) apoptotic pathways. With respect to cellular involvements, fibroblasts are the most well-studied cell population. However, it is unclear whether the fibroblast is the causative cell; they are modulated by other cell populations in wound repair, such as keratinocytes and macrophages. This review presents a detailed account of individual phases of the healing process and how they may potentially be implicated in aberrant raised scar formation, which may help in clarifying the mechanisms involved in keloid disease pathogenesis. PMID- 20002896 TI - The effect of epigallocatechin-3-gallate, a constituent of green tea, on transforming growth factor-beta1-stimulated wound contraction. AB - Dermal fibrosis, or scarring, following surgical incisions, traumatic wounds and burns presents a major clinical burden. Transforming growth factor (TGF)-beta1 is a major factor known to stimulate fibroblast proliferation, collagen production, and the differentiation of fibroblast to myofibroblast promoting wound contraction. Furthermore, excessive or prolonged TGF-beta1 has been shown to be associated with scarring. Green tea contains high amounts of polyphenols with the major polyphenolic compound being epigallocatechin-3-gallate (EGCG). EGCG has been shown to be anti-inflammatory, anti-oxidant, and may improve wound healing and scarring, though its precise effect on TGF-beta1 remains unclear. This study aimed at determining the effect of EGCG on TGF-beta1 collagen contraction, gene expression and the differentiation of fibroblast to myofibroblast. EGCG appears to affect the role that TGF-beta1 plays in fibroblast populated collagen gel contraction and this seems to be through both myofibroblast differentiation and connective tissue growth factor gene expression and reduces the expression of collagen type I gene regulation. PMID- 20002897 TI - Dexamethasone reduces mitomycin C-related inflammatory cytokine expression without inducing further cell death in corneal fibroblasts. AB - The purpose of this study was to investigate the effect of dexamethasone (DEX) on mitomycin C (MMC)-induced inflammatory cytokine expression in corneal fibroblasts. Primary human corneal fibroblasts were treated with MMC, dexamethasone, or in combination. Morphological changes and cell growth were documented using phase-contrast microscopy and PicoGreen assay, respectively. Cell apoptosis was evaluated by annexin V/propidium iodide staining, whereas viability was tested by the live/dead assay and analyzed by flow cytometry. The relative expression of interleukin-8 and monocyte chemoattractant protein-1 was investigated with quantitative real-time polymerase chain reaction and enzyme linked immunosorbent assay. Mitogen-activated protein kinase activation and mitogen-activated protein kinase phosphatase-1 expression were documented by Western blot analysis. We found that MMC induced corneal fibroblast elongation, apoptosis, and retarded cell growth, whereas DEX did not significantly alter cell morphology or viability. The combination of DEX and MMC did not induce additional apoptosis and cell death. DEX dose dependently down-regulated basal and MMC induced interleukin-8 and monocyte chemoattractant protein-1 mRNA expression and protein secretion. DEX attenuated MMC-induced p38 and Jun N-terminal kinases activation and up-regulated expression. These suggested that DEX may inhibit MMC induced interleukin-8 and monocyte chemoattractant protein-1 by up-regulating MKP 1 expression, which subsequently deactivated p38 and Jun N-terminal kinases activation. Combined MMC and DEX treatment may facilitate corneal wound healing. PMID- 20002898 TI - Alteration in cellular morphology, density and distribution in rat vocal fold mucosa following injury. AB - The vocal fold mucosa plays an important role in voice production. Its cellular composition and density frequently change under various pathological conditions, often contributing to altered extracellular matrix production, tissue viscoelasticity, and voice quality. In this study, cellular changes in the rat mucosa following a unilateral stripping injury were investigated and analyzed semi-quantitatively. Distinctive and sequential changes in cellular morphology, composition, and density were observed in the mucosa post-injury. Cellular recruitment was a major event during the early stage of injury and reached its peak level by day 5 post-injury. Several types of cells, including neutrophil like cells, epithelial cells, and fibroblast-like cells, were sequentially recruited. The sequential emergence of reactive cell populations following injury and subsequent reconstruction of the mucosa suggests their involvement in vocal fold tissue repair and scar formation processes. PMID- 20002899 TI - An in silico approach to the analysis of acute wound healing. AB - The complex interactions that characterize acute wound healing have stymied the development of effective therapeutic modalities. The use of computational models holds the promise to improve our basic approach to understanding the process. By modifying an existing ordinary differential equation model of systemic inflammation to simulate local wound healing, we expect to improve the understanding of the underlying complexities of wound healing and thus allow for the development of novel, targeted therapeutic strategies. The modifications in this local acute wound healing model include: evolution from a systemic model to a local model, the incorporation of fibroblast activity, and the effects of tissue oxygenation. Using these modifications we are able to simulate impaired wound healing in hypoxic wounds with varying levels of contamination. Possible therapeutic targets, such as fibroblast death rate and rate of fibroblast recruitment, have been identified by computational analysis. This model is a step toward constructing an integrative systems biology model of human wound healing. PMID- 20002900 TI - Folic acid in the prevention of neural tube defects: awareness among laywomen and healthcare providers in Japan. AB - It is known that neural tube defects are folic acid preventable congenital anomalies. We investigated to what extent this information was disseminated among laywomen and healthcare providers. Questionnaire studies were conducted twice, in 2002 and 2007, for four groups of laywomen and seven groups of healthcare providers in Japan regarding awareness, folic acid supplements and healthy diets. Awareness among laywomen was less than 20%, except for families who had experience with spina bifida in 2002, and 5 years later only pregnant women showed a significant increase in awareness. Awareness among healthcare providers varied from 12 to 76%, depending on their profession, and this proportion increased in five of the seven groups in 2007. The majority of laywomen obtained their information from mass media, while the majority of healthcare providers received information through media for professionals. Laywomen who used folate supplements and healthcare providers who recommended them were initially fewer than 25 and 37%, respectively. Five years later, however, pregnant women who used folic acid supplements increased from 9.1 to 43.1%. As awareness among non pregnant laywomen and some healthcare providers is considerably low, information should be presented repeatedly to these groups. The difficulty in getting women to consume folic acid supplements is an argument for the government to require folic acid fortification of grains so that the prevention of neural tube defects can be maximized. PMID- 20002901 TI - Development of olfactory epithelium in the human fetus: scanning electron microscopic observations. AB - AIMS: Human olfactory epithelium becomes functional at birth, but prenatal development remains unclear. In the present study, we aimed to clarify the development of human olfactory epithelium using scanning electron microscopy (SEM). METHODS: The development of human olfactory epithelium was observed in 24 externally normal fetuses, which were formalin-fixed and long-preserved, with a crown-rump length (CRL) of 102-336 mm (gestational week 14-38). The olfactory mucosa in the superior wall of the nasal septum near the choana were dissected and observed under SEM. We examined the number of olfactory vesicles per unit area, diameter of olfactory vesicles, and number and length of cilia on olfactory vesicles. RESULTS: At circa (ca) CRL 100 mm (ca 14 weeks), olfactory epithelium displayed several olfactory vesicles with 1-2 short cilia per unit area. At ca CRL 150 mm (ca 18 weeks), olfactory vesicles were present in small clusters, and cilia were longer. At CRL lager than 225 mm (ca 26 weeks), olfactory vesicles became located separately from each other, while length and number of cilia per olfactory vesicle were further increased. CONCLUSION: The present findings suggest that fetal olfactory epithelium becomes morphologically almost the same as that in adults in late gestation, much later than previously thought. PMID- 20002902 TI - Restraint-induced maternal stress and alteration of ossification in mouse fetuses. AB - Jcl:ICR pregnant mice were immobilized for 120 minutes from days 8-12 of gestation, and their fetuses were examined for skeletal features on day 18 of gestation. In the stressed group, decreased maternal bodyweight gain and lower fetal weight were noted. In this group, the incidences of segmentation defects, fused ribs, absent lumbar vertebrae and full supernumerary ribs were increased in fetuses. In addition, fusion of the basi- and ex-occipital bones was frequently observed in this group (12.9%). This finding was seen at an incidence of 1.4% in the control group, usually in newborns during the ossification process of the occipital bone. Therefore, the fused basi- and ex-occipital bones were considered to be due to altered ossification, but not to be an abnormality. In summary, immobilization of Jcl:ICR mice during the period of fetal organogenesis induced altered ossification of the occipital bones as well as some abnormalities and supernumerary ribs. PMID- 20002903 TI - Acrania/encephalocele sequence (exencephaly) associated with 92,XXXX karyotype: early prenatal diagnosis at 9(+5) weeks by 3D transvaginal ultrasound and coelocentesis. AB - A 27-year-old pregnant woman was diagnosed by 3D transvaginal ultrasound as carrying a fetus of 9(+5) weeks gestation affected by acrania/encephalocele (exencephaly) sequence. A 2D transvaginal ultrasound-guided aspiration of 5 mL of extra-coelomic fluid was performed under cervical block before uterine suction. Conventional cytogenetic analysis demonstrated a 92,XXXX karyotype. Transvaginal 2D ultrasound-guided coelocentesis for rapid karyotyping can be proposed to women who are near to miscarriage or in cases where a prenatal ultrasound diagnosis of congenital anomaly is performed at an early stage of development. Genetic analysis can be performed using traditional cytogenetic analysis or can be aided by fluorescence in situ hybridization (FISH). Coelocentesis may become an integral part of first trimester armamentarium and may be clinically useful in the understanding of the pathogenesis of early prenatally diagnosed congenital anomalies. PMID- 20002904 TI - Prenatal diagnosis of persistent cloaca. AB - We report four cases of persistent cloaca diagnosed at 32-33 weeks of gestation. In cases of persistent cloaca, serial prenatal ultrasonography shows transient fetal ascites, enlarged cystic structures arising from the fetal pelvis. Our four cases of persistent cloaca were diagnosed prenatally. Persistent cloaca should be considered in any female fetus presenting with hydronephrosis and a large cystic lesion arising from the pelvis as assessed by ultrasound and magnetic resonance imaging. Neither pulmonary hypoplasia nor severe oligohydramnios were found in any of our four cases, and they each had a good prognosis. Prenatal diagnosis allows time for parental counseling and delivery planning at a tertiary care center for neonatal intensive care and pediatric surgery. PMID- 20002905 TI - Thoracic kidney associated with congenital diaphragmatic hernia. AB - We report three cases of ectopic thoracic (or superior ectopic) kidney; one in a neonate and two in 6-month-old children, associated with congenital diaphragmatic hernia. In all cases the diagnosis was made during surgery and confirmed by intravenous pyelography, sonography and magnetic resonance imaging in the postoperative period. Because of the rarity of this condition we report these cases together with a wide review of the published reports. PMID- 20002906 TI - Fetal upper limb amelia with increased nuchal translucency. PMID- 20002907 TI - Terminology of developmental abnormalities in common laboratory mammals (version 2). AB - This update (Version 2) of the Terminology of Developmental Abnormalities in Common Laboratory Mammals (Version 1) incorporates improvements and enhancements to both content and organization of the terminology to enable greater flexibility in its application, while maintaining a consistent approach to the description of findings. The revisions are the result of an international collaboration among interested organizations, advised by individual experts and the outcomes of several workshops. The terminology remains organized into tables under the broad categories of external, visceral, and skeletal observations, following the manner in which data are typically collected and recorded in developmental toxicity studies. This arrangement of the tables, as well as other information provided in appendices, is intended to facilitate the process of specimen evaluation at the laboratory bench level. Only the commonly used laboratory mammals (i.e. rats, mice, rabbits) are addressed in the current terminology tables. The inclusion of other species that are used in developmental toxicity testing, such as primates, is considered outside the scope of the present update. Similarly, categorization of findings as, for example, 'malformation' or 'variation' remains unaddressed, in accordance with the overall principle that the focus of this document is descriptive terminology and not diagnosis or interpretation. The skeletal terms have been augmented to accommodate cartilage findings. PMID- 20002908 TI - Evaluation of PCR assays for quantifying seed-borne infection by Fusarium and Microdochium seedling blight pathogens. AB - AIMS: To evaluate competitive PCR assays for quantifying seed-borne Microdochium and Fusarium seedling blight pathogen DNA and to determine test and year repeatability and sources of variability. METHODS AND RESULTS: Relationships between DNA and plate counts were significant for Fusarium and Microdochium seedling blight pathogens in 152 seed batches from 3 years. Coefficient of determinations, however, differed greatly (Fusarium; R(2) = 0.25, P = 0.029, Microdochium; R(2) = 0.73, P < 0.001). Significant differences between years were observed in the regression slopes for Microdochium. Pathogen DNA quantified in 16 extractions after sampling was highly correlated to results following storage for 1-2 years (R(2) > 0.90). Residual maximum likelihood analysis showed that the least and greatest variance components of the testing procedure were DNA extraction subsampling and PCR assay respectively. CONCLUSIONS: Amount of pathogen DNA is a useful estimator of seed batch contamination for Microdochium but not Fusarium seedling blight pathogens. Although reproducible over time, improvements to the testing procedure should focus on repeated PCR amplifications to reduce assay variability. SIGNIFICANCE AND IMPACT OF THE STUDY: Replacing plate counts with competitive PCR for determining the severity of seed batch contamination is feasible in areas where Microdochium seedling blight pathogens predominate. PMID- 20002909 TI - Characterization of some culture factors affecting oxalate degradation by the mycoparasite Coniothyrium minitans. AB - AIMS: To find possible approaches to utilize the mechanism of oxalate degradation by Coniothyrium minitans (Cm) in controlling the plant pathogen Sclerotinia sclerotiorum (Ss). METHODS AND RESULTS: Differences in oxalate degradation by different Cm strains and effects of the initial oxalate concentration, ambient pH and nutrient factors on mycelial growth and oxalate degradation by Cm were studied in shaken cultures. Results showed that two wild-type Cm strains, Chy-1 and ZS-1, did not differ in oxalate degradation in modified potato dextrose broth (mPDB) amended with oxalic acid (OA). Cm could grow in mPDB amended with sodium oxalate (SO-mPDB) at pH 6.5 or with ammonium oxalate (AO-PDB) at pH 6.2, but oxalate degradation was very low; oxalate degradation was greatly enhanced in SO- or AO-mPDB with pH being lowered to 2.8-2.9. Similarly, oxalate degradation was higher than 90% in OA-amended mPDB at pH 4.4 but was reduced to be <22% at pH 7.0. Five carbon sources and three nitrogen sources investigated and nutrients from mycelia and sclerotia of Ss were favorable for the growth of Cm and OA degradation by Cm. CONCLUSIONS: Cm can degrade oxalate under acidic pH. Exudates from mycelia or sclerotia of Ss may serve as nutrients for Cm mycelial growth and degradation of oxalate secreted by Ss. SIGNIFICANCE AND IMPACT OF THE STUDY: The finding of oxalate degradation laid a foundation for mining-related genes in Cm for engineering plant resistance against Ss. Elucidation of the importance of acidic pH and nutrients from Ss in oxalate degradation by Cm will help to understand the interaction between Cm and Ss. PMID- 20002910 TI - Validation of the dorsal air pouch model to predict and examine immunostimulatory responses in the gut. AB - AIMS: To validate the use of the air pouch system to predict and examine early immune responses induced by the presumptive probiotics Lactobacillus paracasei subsp. paracasei B112, DC205, DC215 and DC412 strains in the gut mucosa. METHODS AND RESULTS: Only the DC412 strain interacted strongly with the cells forming the air pouch lining tissue and induced early innate immune responses such as polymorphonuclear (PMN) cell recruitment, phagocytosis and tumour necrosis factor alpha (TNF-alpha) production that equal the respective responses induced by the probiotic Lactobacillus acidophilus NCFB 1748. The strains exhibiting strong immunoregulatory activity in the air pouch also interacted strongly with the gut associated lymphoid tissue (GALT). The strain DC412 exerts its effect on the intestine through stimulation of Toll-like receptor (TLR)2/TLR4-mediated signalling events leading to secretion of a certain profile of cytokines in which gamma interferon (IFN-gamma), TNF-alpha, interleukin (IL)-6 and IL-10 are included. The probiotic Lact. acidophilus NCFB 1748 induces the same cytokine profile in addition to IL-12B, and this response is potentially mediated by the synergy of TLR2 and TLR9. CONCLUSION: The strain DC412 possesses the in vitro and in vivo characteristics of a probiotic micro-organism. SIGNIFICANCE AND IMPACT OF THE STUDY: The dorsal mouse or rat air pouch may be used as an alternative and rapid method for the initial discrimination and selection of potential probiotic Lactobacillus strains. PMID- 20002911 TI - Characterization of reaerosolization from impingers in an effort to improve airborne virus sampling. AB - AIMS: To assess the impact of reaerosolization from liquid impingement methods on airborne virus sampling. METHODS AND RESULTS: An AGI-30 impinger containing particles [MS2 bacteriophage or 30-nm polystyrene latex (PSL)] of known concentration was operated with sterile air. Reaerosolized particles as a function of sampling flow rate and particle concentration in the impinger collection liquid were characterized using a scanning mobility particle sizer. Reaerosolization from the impinger was also compared to that from a BioSampler. Results show that reaerosolization increases as flow rate increases. While the increased particle concentration in the impinger collection liquid leads to an increase in the reaerosolization of PSL particles, it does not necessarily lead to an increase in the reaerosolization of virus particles. Reaerosolization of virus particles begins to decrease as the particle concentration in the impinger collection liquid rises above 10(6) PFU ml(-1). This phenomenon results from aggregation of viral particles at high concentrations. Compared with micron-sized particles, nanosized virus particles are easier to aerosolize because of reduced inertia. Reaerosolization from the BioSampler is demonstrated to be significantly less than that from the impinger. CONCLUSIONS: Reaerosolization from impingement sampling methods is a mode of loss in airborne virus sampling, although it is not as significant a limitation as the primary particle size of the aerosol. Utilizing a BioSampler coupled with short sampling periods to prevent high accumulative concentrations can minimize the impact of reaerosolization. SIGNIFICANCE AND IMPACT OF THE STUDY: This study confirms reaerosolization of virus particles to be a mode of loss in impingement sampling and identifies methods to minimize the loss. PMID- 20002912 TI - Synergistic mechanisms of retigeric acid B and azoles against Candida albicans. AB - AIMS: To clarify the underlying synergistic antifungal mechanisms of retigeric acid B (RAB) in combination with azoles against Candida albicans. METHODS AND RESULTS: Increased accumulation of rhodamine 123 in C. albicans was measured by both spectrophotometric method and flow cytometry. The inhibitory properties to the drug efflux of C. albicans were determined spectrophotometrically. The decreased cellular ergosterol synthesis was measured using its unique spectrophotometric absorbance profile, and the downregulation expression levels of CDR1 and ERG11 were detected by real-time reverse transcription polymerase chain reaction. Transmission electron microscopy investigation found the wrinkled cell membrane and the impaired cell wall. CONCLUSIONS: RAB synergizes the antifungal effect of azoles against C. albicans by inhibiting efflux pump activity, targeting the ergosterol biosynthesis pathway and increasing the fluidity for the resulted ergosterol depletion. SIGNIFICANCE AND IMPACT OF THE STUDY: Investigating the mechanism of the synergy between RAB and azoles against C. albicans will help us to uncover the antifungal roles of this lichen-derived triterpene acid and find its possible clinical applications in overcoming fungal resistance. PMID- 20002913 TI - Proteomic analysis of outer membrane proteins of Edwardsiella tarda. AB - AIMS: The purpose of this study was to identify outer membrane proteins (OMPs) of Edwardsiella tarda. METHODS AND RESULTS: The OMPs from a virulent strain of E. tarda (ET-7) was extracted using lauroyl sarcosine method. The OMPs were analysed by two-dimensional polyacrylamide gel electrophoresis (2D-PAGE), and protein spots were identified using matrix assisted laser desorption/ionization-time-of flight mass spectrometry. A total of 21 proteins were identified from 24 protein spots observed on the 2D-PAGE gel. These proteins were identified as GroEL, antigenic proteins, ABC transporters, elongation factors, OmpA, PTSINtr with GAF domain, catalase C, glycolytic enzymes, DnaJ, transcriptional regulator, proteins mraZ and ccdA. Subcellular localizations, beta-barrel OMPs and lipoproteins of identified proteins were predicted using PSORTb, PRED-TMBB and LipoP1.0 programme. CONCLUSIONS: Identification, localization and possible functions of OMPs of E. tarda were studied. SIGNIFICANCE AND IMPACT OF THE STUDY: These proteins could be used for development of novel drug targets, diagnostics or vaccine against edwardsiellosis. PMID- 20002914 TI - [Journal-club]. PMID- 20002915 TI - [Scaling erythemas in an infant]. PMID- 20002916 TI - [The star plasty--modification of the purse string suture]. PMID- 20002920 TI - The enduring merits of gold. PMID- 20002921 TI - Dental erosion. PMID- 20002922 TI - Conservative and esthetic cast gold fixed partial dentures-inlay, onlay, and partial veneer retainers, custom composite pontics, and stress-breakers: part I: fundamental design principles. AB - Although in today's dental world implant restorations are considered the standard of care in the replacement of missing teeth, clinical contraindications and patient nonacceptance of implant placement can be encountered. Several scenarios are discussed here in which a single missing tooth can be restored with conservative fixed partial dentures (FPD) that employ cast gold retainers; each with a customized design in order to preserve tooth structure, maintain esthetics, and provide a long-term prognosis. The abutment teeth are prepared for conservative partial coverage restorations by using Brasseler burs (Brasseler USA, Savannah, GA, USA). Impressions are taken of the preparations, along with any retentive features, utilizing either the Vented Pin Channel technique or the Shooshan Plastic Pin technique. The latter technique utilizes Kodex twist drills and corresponding impression pins (Coltene Whaledent Inc., Mahwah, NJ, USA). The conservative FPD with non-rigid connectors is fabricated by using type III gold alloy. The pontic cage portion is chemically prepared utilizing the Panavia F2.0 cement kit (Kuraray America Inc., Houston, TX, USA) or other dual-polymerizing resin cement and restored with any type of direct composite resin material. A palette of opaquers and tints are used for chairside characterization of the esthetic pontic facing. The final polish of the pontic is completed by using FlexiDisc and FlexiBuff discs (Cosmedent Inc., Chicago, IL, USA). CLINICAL SIGNIFICANCE In cases where an implant restoration is contraindicated for replacement of a single tooth, a semi-precision FPD is a conservative, functional, and esthetic alternative. PMID- 20002923 TI - Conservative and esthetic cast gold fixed partial dentures-inlay, onlay, and partial veneer retainers, custom composite pontics, and stress breakers: part II: utilization of additional retentive features and fabrication of custom pontic facings. AB - Although in today's dental world implant restorations are considered the standard of care in the replacement of missing teeth, clinical contraindications and patient nonacceptance of implant placement can be encountered. Several scenarios are discussed here in which a single missing tooth can be restored with conservative fixed partial dentures (FPD) that employ cast gold retainers; each with a customized design in order to preserve tooth structure, maintain esthetics, and provide a long-term prognosis. The abutment teeth are prepared for conservative partial coverage restorations by using Brasseler burs (Brasseler USA, Savannah, GA, USA). Impressions are taken of the preparations, along with any retentive features, utilizing either the Vented Pin Channel technique or the Shooshan Plastic Pin technique. The latter technique utilizes Kodex twist drills and corresponding impression pins (Coltene Whaledent Inc., Mahwah, NJ, USA). The conservative FPD with non-rigid connectors is fabricated by using type III gold alloy. The pontic cage portion is chemically prepared utilizing the Panavia F2.0 cement kit (Kuraray America Inc., Houston, TX, USA) or other dual-polymerizing resin cement and restored with any type of direct composite resin material. A palette of opaquers and tints are used for chairside characterization of the esthetic pontic facing. The final polish of the pontic is completed by using FlexiDisc and FlexiBuff discs (Cosmedent Inc., Chicago, IL, USA). CLINICAL SIGNIFICANCE In cases where an implant restoration is contraindicated for replacement of a single tooth, a semi-precision FPD is a conservative, functional, and esthetic alternative. PMID- 20002924 TI - Commentary: conservative and esthetic cast gold fixed partial dentures-inlay, onlay, and partial veneer retainers, custom composite pontics, and stress breakers: parts I and II. PMID- 20002925 TI - Effect of light irradiation on tooth whitening: enamel microhardness and color change. AB - The aim of this study was to evaluate the influence of light exposure associated with 35% hydrogen peroxide (Pola Office, SDI, Melbourne, Vic., Australia) or 15% hydrogen peroxide (BriteSmile, Discus, Culver City, CA, USA) on the microhardness and color changes of bovine enamel. Experimental groups were Britesmile + Light (BL) (15% hydrogen peroxide + plasm arc; 4 x 20 minutes), Britesmile + No Light (BN) (BL, no light), Pola office + Light (PL) (35% hydrogen peroxide + LED; 4 x 8 minutes), and Pola office + No light (PN) (PL, no light). Color changes (DeltaE) and the CIELAB (Commission Internationale de l' Eclairage, L* a* b* color system) parameters (L*, a*, and b*) were assessed with a spectrophotometer before (B), immediately (A), 1 day and 7 days after bleaching. The microhardness was measured before (B) and after (A), the obtained data were submitted to a two-way analysis of variance, and DeltaE were submitted to t-test for each period. Only Pola Office, in which the peroxide is associated with the light, improved DeltaE when evaluated immediately after bleaching (p < 0.001). Light exposure did not influence DeltaE after 1 day or 7 days for either bleaching system. The enamel microhardness was not altered after bleaching for BriteSmile. However, enamel microhardness was reduced after bleaching for Pola Office, 283 MPa (+/-21) and 265 MPa (+/-27), respectively. It was concluded that these two bleaching systems were efficient regardless of the light systems used. However, the 35% hydrogen peroxide altered the enamel microhardness. CLINICAL SIGNIFICANCE Enamel microhardness was affected by a 35% hydrogen peroxide in-office bleaching therapy. Moreover, the in-office bleaching outcome was not improved by using the light associated with systems tested in this study. (J Esthet Restor Dent 21:387 396, 2009). PMID- 20002926 TI - Commentary: effect of light irradiation on tooth whitening: enamel microhardness and color change. PMID- 20002927 TI - Effect of surface sealants on marginal microleakage in Class V resin composite restorations. AB - PURPOSE: Surface sealants may reduce or avoid problems related to the marginal interface. The aim of this study was to evaluate the microleakage in resin composite Class V restorations sealed with an adhesive system (Xeno III [Dentsply, Konstanz, Germany]), a sealant for exposed dentin (Seal & Protect [Dentsply]), and two surface sealants (Fortify [Bisco, Lombard, IL, USA]; Optiguard [Kerr, Orange, CA, USA]). MATERIALS AND METHODS: Fifty cavities with margins in enamel were prepared on crowns of bovine teeth and restored with an adhesive system (Prime and Bond NT/Dentsply) and resin composite (Esthet X/Dentsply). They were separated into four groups for the application of the surface sealants, and a control group (without surface sealing). Thermal cycling with baths of 5 degrees C +/- 2 degrees C and 55 degrees C +/- 2 degrees C were performed in 600 cycles. The teeth were made impermeable, except for a 3-mm window around the restoration and immersed in a 50% silver nitrate solution for 8 hours. The crown was sectioned longitudinally and the cuts were analyzed by three independent evaluators, using a stereoscopic loupe with 10x magnification that attributed representative scores. Agreement among the examiners was evaluated by the Kappa test. RESULTS: The Kruskal-Wallis test and Dunn Method showed that there was significant difference between the Control and Seal & Protect groups. The Seal & Protect group presented the lowest degree of microleakage, followed by the Optiguard and Xeno III groups. The highest scores were obtained in the Control group. CONCLUSION: The sealant materials evaluated presented different rates of effectiveness, and Seal & Protect was the most effective in decreasing the degree of marginal microleakage. CLINICAL SIGNIFICANCE Although surface sealants or covering agents have been used to diminish microleakage, they may present different rates of effectiveness with regard to reducing the degree of marginal leakage. (J Esthet Restor Dent 21:397-406, 2009). PMID- 20002928 TI - Commentary: effect of surface sealants on marginal microleakage in Class V resin composite restorations. PMID- 20002929 TI - Effect of bleaching on staining susceptibility of resin composite restorative materials. AB - STATEMENT OF THE PROBLEM: Effect of bleaching procedures on staining susceptibility of resin restorative materials is still questionable. PURPOSE: The aim of this study was to evaluate the staining susceptibility of restorative materials bleached with 20% carbamide peroxide home bleaching agent and subsequently immersed in coffee and tea. MATERIALS AND METHODS: Forty-two disk shaped specimens were fabricated for each of the resin composites (Filtek Supreme XT [3M ESPE, St. Paul, MN, USA], Ceram-X Mono [Dentsply, Konstanz, Germany], and Aelite All Purpose Body [BISCO, Inc., Shaumburg, IL, USA]). The baseline color values were measured with a spectrophotometer. The specimens of each restorative material were randomly divided into two groups (N = 21). While the first group specimens were stored in distilled water (nonbleaching group-control), bleaching agent (Opalescence PF 20% [Ultradent Poducts, South Jordan, UT, USA]) was applied on the top surface of each specimen of the second group (bleaching group). After color change values were measured, the specimens were randomly divided into three subgroups (N = 7) according to the staining solutions. The color change values (DeltaE*ab) were calculated and the data were subjected to analysis of variance. Statistical significance was declared if the p value was 0.05 or less. RESULTS: There was no statistically significant difference within each restorative material's DeltaE*ab values after bleaching (p = 0.714). Also, the staining solutions did not cause a statistically significant difference between DeltaE*ab values of bleaching compared with nonbleaching groups (p = 0.146). Significant interaction was found only between restorative materials and staining solutions (p = 0.000). CONCLUSION: Bleaching of the tested resin composites did not increase their susceptibility to extrinsic staining in vitro. CLINICAL SIGNIFICANCE Bleaching did not affect staining susceptibility of the tested resin composite restorative materials. (J Esthet Restor Dent 21:407-415, 2009). PMID- 20002930 TI - Commentary: effect of bleaching on staining susceptibility of resin composite restorative materials. PMID- 20002931 TI - Critical appraisal: bonding to fluorosed tooth structure. AB - In many continuing education lectures, I frequently have been asked the question "When bonding to fluorosed teeth, should I do anything differently?" As the prevalence of fluorosis has increased, this question has become more and more clinically relevant. Unfortunately, there has not been extensive research in this area, but there has been some. Recent Ask the Expert pieces in the Journal have addressed the issue, and this Critical Appraisal covers the recent research on bonding of resin-based materials to fluorosed enamel and dentin in greater detail. PMID- 20002932 TI - Talking with Patients. Featured topic: fluoride. PMID- 20002933 TI - Mixed infections may promote diversification of mutualistic symbionts: why are there ineffective rhizobia? AB - While strategy variation is a key feature of symbiotic mutualisms, little work focuses on the origin of this diversity. Rhizobia strategies range from mutualistic nitrogen fixers to parasitic nonfixers that hoard plant resources to increase their own survival in soil. Host plants reward beneficial rhizobia with higher nodule growth rates, generating a trade-off between reproduction in nodules and subsequent survival in soil. However, hosts might not discriminate between strains in mixed infections, allowing nonfixing strains to escape sanctions. We construct an adaptive dynamics model of symbiotic nitrogen-fixation and find general situations where symbionts undergo adaptive diversification, but in most situations complete nonfixers do not evolve. Social conflict in mixed infections when symbionts face a survival-reproduction trade-off can drive the origin of some coexisting symbiont strategies, where less mutualistic strains exploit benefits generated by better mutualists. PMID- 20002936 TI - Another key molecule for pathogenesis of hepatocellular carcinoma. PMID- 20002937 TI - Adding serum sodium to model for end-stage liver disease: identifying those most at risk. PMID- 20002938 TI - Moderate acute cholecystitis: to cut now or to cut later. PMID- 20002939 TI - Endoscopic ultrasound-fine needle aspiration for the diagnosis of lymphoma: are we there yet? PMID- 20002940 TI - Epidemiology of colorectal cancer in Asia. AB - The prevalence of colorectal cancer is increasing in Asia. However, the age standardized rate has reached a plateau in some countries. Some studies have shown a male predominance difference and increasing risk in the elderly, but not in the younger population. 'Right shifting' of colorectal cancer, not accountable by difference in age or the indications for endoscopic examination, has also been noted. Westernized diet is associated with colorectal cancer, but controversy remains on how it causes colorectal cancer. Alcohol consumption, obesity, diabetes mellitus, consumption of red and processed meat and cigarette smoking are linked to bowel cancer epidemiologically. Only high dietary calcium has a consistent negative (or 'protective') effect. The efficacy of fish oil, vitamin D, soy, phytoestrogens, folate, methionine, riboflavin and vitamin B6 has not been established. Aspirin and non-steroidal anti-inflammatory drugs use decrease risk of colorectal cancer after 5-10 years of use. There is no evidence for a detrimental effect of proton pump inhibitors or benefit of statins in colorectal cancer. In conclusion, there is a rising trend and prevalence of colorectal cancer in Asia. Dietary modification or supplementation may not be effective in preventing colorectal cancer. Surveillance of colorectal cancer in high-risk groups, according to current recommendation, is probably most effective. PMID- 20002941 TI - Proliferative drive and liver carcinogenesis: too much of a good thing? AB - There have been innumerable studies published in the attempt to identify gene expression signatures in hepatocellular carcinoma (HCC). When all the regulators and targets of the differentially expressed genes are analyzed from larger studies, the most striking theme is upregulation of mitosis-promoting and cell proliferation genes in HCC compared with 'liver-specific gene clusters' in non tumorous tissue. A major limitation of expression profiling is that it only provides a 'snapshot' of what is an evolving process and thus cannot distinguish the differences in gene expression that are primary effectors of dysregulated growth from those that represent downstream consequences. The development of HCC in a chronically diseased liver, often referred to as hepatocarcinogenesis, is a multistep process characterized by the progressive accumulation and interplay of genetic alterations causing aberrant growth, malignant transformation of liver parenchymal cells, followed by vascular invasion and metastasis. This review will discuss HCC precursor lesions, draw on the 'proliferation cluster' genes highlighted from HCC expression profiling studies, relate them to a selection of regulatory networks important in liver regeneration, cell cycle control and their potential significance in the pathogenesis of HCC or primary liver cancer. PMID- 20002942 TI - Gastrointestinal: acute haemorrhage from a Dieulafoy lesion within a gastric diverticulum managed endoscopically. PMID- 20002943 TI - Gastrointestinal: Eosinophilic enteritis manifesting as brown-pigmented duodenal ulcers. PMID- 20002944 TI - Hepatobiliary and pancreatic: papillary cholangiocarcinoma. PMID- 20002947 TI - Nutrition in older people. PMID- 20002949 TI - Communication competence within dietetics: dietitians' and clients' views about the unspoken dialogue--the impact of personal presentation. AB - BACKGROUND: Although client communications are affected by clients' assumptions about professionals' characteristics drawn from dress attire, little is known about how this dialogue operates in dietetics. The present study aimed to describe how dietitians and their clients interpret this dialogue and to explore the implications for practice. METHODS: A purposive quota sample of dietitians (n = 46) from 21 health services in one state of Australia and a quota of their adult patients (n = 34) were interviewed about dietitians' nutrition education roles. Semi-structured interviews were transcribed and identified themes developed into a questionnaire to survey Australian dietitians. Analysis used frequencies and nonparametric statistics (P <0.05). Triangulation of the results obtained from the studies revealed a strong agreement between data sources. RESULTS: Dietitians' dress attire was perceived as a key source of nonverbal communications by dietetics clients. This was recognised by 75% of the 256 dietitians who were surveyed nationally. Dietitians favoured a professional style (i.e. skirt or slacks, with top). Many clients rejected formal dress (i.e. suit, high heels) as being a potential communication barrier. Some clients viewed dietitians' bodily size/shape as a role model. Implications of dietitians' presentation (i.e. how you look) were important to both clients and dietitians. CONCLUSION: Dress style is implicated in nonverbal communication dialogues between the dietitian and client. As a matter of competence and to maintain congruency in communication, dietitians should be aware of their clients' preferences for formality of dress, and conduct their attire accordingly. PMID- 20002950 TI - Validity, reliability and acceptability of the Imperial Nutritional Screening System (INSYST): a tool that does not require the body mass index. AB - BACKGROUND: Nutritional screening tools are central to identifying malnourished patients, but their efficacy is often reduced as a result of difficulties in obtaining height for body mass index (BMI) calculations. The present study aimed to evaluate the validity, reliability and acceptability of the Imperial Nutritional Screening System (INSYST); a tool that does not require the BMI. METHODS: Patients were screened by the researcher within 72 h of admission using INSYST I & II, Malnutrition Universal Screening Tool (MUST) and Mini Nutritional Assessment (MNA), including taking height and weight. Routine INSYST data, completed by nursing staff, were subsequently collected. At risk and malnourished patients were combined for statistical analysis. Inter-tool and inter-rater agreement (kappa, kappa) was evaluated. Sensitivity and specificity were calculated. Nurses were timed using INSYST. Acceptability, including ease and speed of use, was evaluated. RESULTS: Kappa (agreement) scores (all P < 0.001) were substantial for INSYST I versus MUST and MNA (kappa = 0.73 and kappa = 0.76, respectively) and moderate for INSYST II (both kappa = 0.53). The sensitivity of INSYST I and II was high (95-100%), whereas specificity was lower (65-83%). The agreement between dietitian and nurse for INSYST I was substantial kappa = 0.77 and that for INSYST II was fair kappa = 0.39 (both P or=18 years (of whom 65% claimed to be 'motivated to eat a healthy diet') indicated that over 40% reported eating at a catering outlet at least once a week. Over half said that they would wish to see information on ingredients and the salt content of menu items at all venues. Preference for information on energy and fat content was less popular and varied in the range 42-56% for energy and 47-59% for fat. It is notable that 43% of respondents said they would welcome information on energy content of menu items in restaurants. CONCLUSION: A significant proportion of consumers wish to see information on the ingredients and nutrition composition on menu items for sale in UK catering outlets. Such information is likely to raise an awareness and understanding of healthy food choices and assist the population in making informed choices about healthy eating. PMID- 20002960 TI - A novel 3-d reconstruction system for the assessment of bifurcation lesions treated by the mini-crush technique. AB - BACKGROUND: Conventional two-dimensional angiography lacks the ability to properly image the true bifurcation geometry, and its percutaneous coronary intervention-induced changes in the clinical setting. METHODS AND RESULTS: A novel three-dimensional reconstruction system was investigated by retrospectively analyzing 39 lesions in 35 consecutive patients with coronary bifurcation disease treated with the mini-crush technique. At baseline, significant correlations were proved between two- and three-dimensional systems in terms of either reference vessel diameter (R(2)= 0.68 and 0.29 for main and side branches, respectively), minimum lumen diameter (R(2)= 0.73 and 0.36), stenosis diameter (R(2)= 0.69 and 0.29), and lesion length (R(2)= 0.48 and 0.58). These results were consistent with those observed after the procedure and at 8-month follow-up. Lesion length was significantly longer with the three-dimensional compared to the two dimensional system for both main and side branches (P < 0.001, and P = 0.007, respectively). CONCLUSIONS: The three-dimensional quantitative reconstruction system may provide accurate evaluation of the complex curvilinear structure of bifurcation lesions when using a double stent technique. PMID- 20002961 TI - Photoperiodic modulation of clock gene expression in the avian premammillary nucleus. AB - The premammillary nucleus (PMM) has been shown to contain a daily endogenous dual oscillation in dopamine (DA)/melatonin (MEL) as well as c-fos mRNA expression that is associated with the daily photo-inducible phase of gonad growth in turkeys. In the present study, the expression of clock genes (Bmal1, Clock, Cry1, Cry2, Per2 and Per3) in the PMM was determined under short (8 : 16 h light/dark cycle) and long (16 : 8 h light/dark cycle) photoperiods relative to changes associated with the diurnal rhythm of DA and MEL. Constant darkness (0 : 24 h light/dark cycle) was used to assess the endogenous response of clock genes. In addition, light pulses were given at zeitgeber time (ZT) 8, 14 and 20 to ascertain whether clock gene expression is modulated by light pulse stimulation and therefore has a daily phase-related response. In the PMM, the temporal clock gene expression profiles were similar under short and long photoperiods, except that Per3 gene was phase-delayed by approximately 16 h under long photoperiod. In addition, Cry1 and Per3 genes were light-induced at ZT 14, the photosensitive phase for gonad recrudescence, whereas the Clock gene was repressed. Gene expression in established circadian pacemakers, the visual suprachiasmatic nucleus (vSCN) and the pineal, was also determined. Clock genes in the pineal gland were rhythmic under both photoperiods, and were not altered after light pulses at ZT 14, which suggests that pineal clock genes may not be associated with the photosensitive phase and reproductive activities. In the vSCN, clock gene expression was phase-shifted depending on the photoperiod, with apexes at night under short day length and during the day under long day length. Furthermore, light pulses at ZT 14 induced the Per2 gene, whereas it repressed the Bmal1 gene. Taken together, the changes in clock gene expression observed within the PMM were unique compared to the pineal and vSCN, and were induced by long photoperiod and light during the daily photosensitive phase; stimuli that are also documented to promote reproductive activity. These results show that Cry1 and Per3 are involved in the photic response associated with the PMM neuronal activation and are coincident with an essential circadian mechanism (photosensitive phase) controlling the reproductive neuroendocrine system. PMID- 20002962 TI - Corticotrophin-releasing factor alters the timing of puberty in the female rat. AB - Puberty is a developmental process that is dependent upon activation of the hypothalamic gonadotrophin-releasing hormone (GnRH) pulse generator. It is well established that the stress neuropeptide, corticotrophin-releasing factor (CRF), has a profound inhibitory action on GnRH pulse generator frequency. Although stress is known to affect the timing of puberty, the role of CRF is unknown. The present study aimed to test the hypothesis that CRF plays a critical role in the timing of puberty. On postnatal day (pnd) 28, female rat pups were chronically implanted with i.c.v. cannulae and received 14 days of administration of either CRF, CRF receptor antagonist (astressin-B) or artificial cerebrospinal fluid via an osmotic mini-pump. A separate group of rats served as nonsurgical controls. As a marker of puberty, rats were monitored for vaginal opening and first vaginal oestrus. Levels of CRF, CRF receptor types 1 and 2 (CRF-R1, CRF-R2) mRNA expression in micropunches of the medial preoptic area (mPOA), hypothalamic paraventricular nucleus (PVN) and arcuate nucleus (ARC) were determined across pubertal development; brain tissue was collected from a naive group of rats on pnd 14, 32, on the day of vaginal opening, and pnd 77 (Adult). Administration of CRF resulted in a delay in the onset of puberty, whereas astressin-B advanced pubertal onset. Additionally, CRF and CRF-R1 mRNA expression was reduced in the mPOA, but not ARC, at puberty. In the PVN, expression of CRF, but not CRF-R1 mRNA, was reduced at the time of puberty. These data support the hypothesis that CRF signalling may play an important role in modulating the timing of puberty in the rat. PMID- 20002963 TI - Effects of photoperiod on kisspeptin neuronal populations of the ewe diencephalon in connection with reproductive function. AB - Kisspeptin (Kiss) is a key regulator of reproductive function in both prepubertal and adult mammals. Its expression appears to vary throughout the year in seasonal species. We aimed to determine the impact of a change of photoperiod on the size of Kiss neuronal populations found in the preoptic area (POA) and arcuate nucleus (ARC) of the ewe brain. Using immunocytochemistry, we first examined the proportion of neurones expressing Kiss, using HuC/D as a neuronal marker, at different time-points after transition from long days (LD; 16 : 8 h light/dark cycle) to short days (SD; 8 : 16 h light/dark cycle). Luteinising hormone (LH) secretion was measured in ovariectomised oestradiol replaced ewes from the month preceding the transition to SD until the sacrifice of the animals at days 0, 45 and 112 from this photoperiodic transition. High LH levels were only observed in animals killed at day 112. The number of Kiss neurones/mm(2) doubled in the caudal ARC at day 112. The percentage of neurones showing Kiss immunoreactivity increased significantly in both the POA and ARC in the day 112 group. In a second experiment, ewes kept in LD received an i.c.v. injection of colchicine 20 h before sacrifice. Colchicine treatment increased the number and the percentage of neurones with Kiss in both the POA and caudal ARC. The data obtained suggest that the increase in Kiss neurones detected in the POA and caudal ARC after transition to SD stemmed from an increase in Kiss synthesis. This up-regulation of Kiss content under the shorter day condition appears to be a late event within the cascade activated by a longer secretion of melatonin, which is a critical factor in switching gonadotrophin-releasing hormone secretion to a breeding season profile. PMID- 20002964 TI - The response of glucose-excited neurones in the ventromedial hypothalamus to decreased glucose is enhanced in a murine model of type 2 diabetes mellitus. AB - Obesity and type 2 diabetes mellitus (T2DM) are associated with dysfunctional insulin signalling and impaired central glucose sensing. Glucose sensing neurones reside in key areas of the brain involved in glucose and energy homeostasis (e.g. ventromedial hypothalamus; VMH). We have recently shown that insulin attenuates the ability of glucose-excited (GE) neurones to sense decreased glucose. We hypothesise that this effect of insulin on VMH GE neurones is impaired during T2DM when insulin signalling is dysfunctional. To test our hypotheses, we used whole cell patch clamp recording techniques to evaluate the effects of insulin on VMH GE neurones in brain slices from wild-type and diabetic (db/db) mice. The effects of decreasing glucose from 2.5 to 0.1 mM on VMH GE neurones were similar in wild-type and db/db mice. However, decreasing glucose from 2.5 to 0.5 mM decreased the action potential frequency, membrane potential and input resistance of VMH GE neurones to a significantly greater extent in db/db versus wild-type mice. Furthermore, insulin (5 nM) blunted the effects of decreased glucose in wild-type, but not db/db mice. These differences in both glucose and insulin sensitivity between wild-type and db/db mice were completely ameliorated by the insulin sensitiser, Compound 2 (300 nM). These data are consistent with our hypothesis that impaired insulin signalling in T2DM sensitises VMH GE neurones to decreased glucose. PMID- 20002965 TI - A comparison of two repeated restraint stress paradigms on hypothalamic-pituitary adrenal axis habituation, gonadal status and central neuropeptide expression in adult male rats. AB - The available evidence continues to illustrate an inhibitory influence of male gonadal activity on the hypothalamic-pituitary-adrenal (HPA) axis under acute stress. However, far less is known about how these systems interact during repeated stress. Because HPA output consistently declines across studies examining repeated restraint, the potential mechanisms mediating this habituation are often inferred as being equivalent, even though these studies use a spectrum of restraint durations and exposures. To test this generalisation, as well as to emphasise a potential influence of the male gonadal axis on the process of HPA habituation, we compared the effects of two commonly used paradigms of repeated restraint in the rodent: ten daily episodes of 0.5 h of restraint and five daily episodes of 3 h of restraint. Both paradigms produced comparable declines in adrenocorticotrophic hormone and corticosterone between the first and last day of testing. However, marked differences in testosterone levels, as well as corticotrophin-releasing hormone (CRH) and arginine vasopressin (AVP) expression, occurred between the two stress groups. Plasma testosterone levels remained relatively higher in animals exposed to 0.5 h of restraint compared to 3 h of restraint, whereas forebrain gonadotrophin-releasing hormone (GnRH) cell counts increased in both groups. AVP mRNA was increased after 3 h, but not after 0.5 h of repeated restraint, in the medial parvicellular paraventricular nucleus and in the posterior bed nucleus of the stria terminalis (BST), and increased with 0.5 h of repeated restraint in the medial amygdala. CRH mRNA was increased after 3 h, but not after 0.5 h of repeated restraint, in the central amygdala and anterior BST. The data obtained illustrate that, despite comparable declines in HPA responses, the pathways recruited for stress adaptation appear to be distinct between restraint groups. Given the extreme sensitivity of limbic AVP to testosterone, and conversely CRH to circulating glucocorticoids, whether differences in endocrine profiles might explain these neuropeptide differences remains to be seen. Nonetheless, the present study provides several new entry points for testing gonadal influences on stress-specific HPA habituation. PMID- 20002966 TI - The role of transcranial Doppler embolic monitoring in the management of intracranial arterial stenosis. AB - Intracranial arterial stenosis (IAS) is thought to be responsible for 8% of all ischemic stroke subtypes. The best medical treatment for this condition is still controversial. Transcranial Doppler (TCD) emboli monitoring may help to guide the treatment by measuring the frequency of microembolic signals (MES). We report a case of IAS where TCD emboli monitoring proved useful in establishing the mechanism of stroke by being artery-to-artery emboli and guiding therapy based on the frequent symptoms and number of MES. PMID- 20002967 TI - MRI evidence of early muscle atrophy in MuSK positive myasthenia gravis. AB - Muscle atrophy, particularly of facial and bulbar muscles, seems to be a relatively common long-term consequence of musclespecific tyrosine kinase myasthenia gravis (MuSK-MG), perhaps reflecting the chronic state of disease or long-term therapy with corticosteroids. We performed magnetic resonance imaging (MRI) to assess muscle wasting in the facial and bulbar muscles in two female MuSK-MG patients, with short duration of symptoms prior to diagnosis and prior to commencement of steroid therapy. The study revealed marked atrophy of temporalis, masseters, and lingual muscles with fatty replacement. MRI evidence of early muscle atrophy in MuSK-MG may indicate that MuSK antibodies per se may predispose to muscle atrophy. PMID- 20002968 TI - Recognition of human emotions from cerebral blood flow changes in the frontal region: a study with event-related near-infrared spectroscopy. AB - BACKGROUNDS AND PURPOSE: The aim of this study is to develop a near-infrared spectroscopy (NIRS)-based system that recognizes pleasant and unpleasant human emotions based on cerebral blood flow (CBF) in order to understand the minds of patients whose brain function is severely impaired. The forehead region is easily accessible to NIRS measurements, whereas the role of the anterior prefrontal cortex (PFC) in the processing of emotion remains to be elucidated. METHODS: Initially, using event-related NIRS we examined changes in oxygenated hemoglobin (oxy-Hb) as an indicator of regional CBF changes, which reflect brain activity directly related to emotions, but not to cognitive operations in the anterior frontal regions, during viewing affective pictures. The event-related potentials (ERPs), systemic blood pressure, and pulse rate were also measured simultaneously. RESULTS: The event-related analysis of changes in oxy-Hb for a 6 s-picture presentation period showed that very unpleasant emotion was accompanied by an increase in oxy-Hb in the bilateral ventrolateral PFCs, while very pleasant emotion was accompanied by a decrease in oxy-Hb in the left dorsolateral PFC. There were no significant differences in either ERPs or autonomic nervous system activities between the two emotional states. CONCLUSION: These findings suggest the possibility of recognizing patients' emotions from CBF changes. PMID- 20002969 TI - Septo-optic dysplasia complicated by infantile spasms and bilateral choroidal fissure arachnoid cysts. AB - BACKGROUND AND PURPOSE: septo-optic dysplasia (SOD) is the triad of optic nerve hypoplasia, panhypopituitarism, and agenesis of septum pellucidum, and has been described previously to be associated with heterotopias and midline interhemispheric cyst. We describe a case of SOD with arachnoid cysts, persistent primary hyperplastic vitreous, and malformations of cortical development. METHODS: case report and review of literature. RESULTS: our patient was found to have SOD, bilateral ventriculomegaly, pachygyria, gray matter heterotopia, bilateral choroidal cysts near the brainstem, and persistent primary hyperplastic vitreous. She later developed infantile spasms and required enucleation of the abnormal eye and cyst fenestration. CONCLUSION: coincidence of seizures, SOD, bilateral choroid fissure cysts, heterotopias, and persistent primary hyperplastic vitreous is a unique constellation. It is unclear whether this represents a new syndrome or SOD spectrum variation. Patients with SOD and arachnoid cysts should be monitored for signs of herniation. PMID- 20002970 TI - Extraparenchymal neurocysticercosis in Albuquerque, New Mexico. AB - BACKGROUND: neurocysticercosis (NCC) prevalence is increasing throughout the United States mainly because of immigration from Latin America. Clinicians may fail to recognize the extraparenchymal disease because they do not consider the diagnosis. METHODS: to analyze neuroimaging and clinical characteristics of extraparenchymal NCC, we retrospectively reviewed all such cases presenting to a major general medical school hospital in the State of New Mexico. RESULTS: eleven (30%) of our 37 cases of NCC diagnosed using standard criteria from 1998 through 2004 had extraparenchymal disease. On neuroimaging, 36% of the patients lacked parenchymal cysts, 64% had intraventricular cysticerci, 64% had subarachnoid cysticerci, and 64% had hydrocephalus due to either basal arachnoiditis or direct obstruction of intraventricular pathways. Lumbar puncture was performed in 6 patients. All had a cerebrospinal fluid (CSF) pleocytosis, none had CSF or blood eosinophilia, and CSF antibody to NCC could be absent while present in serum. Response to treatment was frequently suboptimal. CONCLUSIONS: extraparenchymal NCC is more frequent than previously thought. Because clinicians outside the Southwest United States are often unfamiliar with NCC as a cause of chronic meningitis, chronic ventriculitis, or hydrocephalus without obvious cysts, the diagnosis of extraparenchymal NCC often depends on the correct interpretation of neuroimaging. PMID- 20002971 TI - Validation of transcranial Doppler with magnetic resonance angiography in acute cerebral ischemia. AB - BACKGROUND AND PURPOSE: Transcranial Doppler (TCD) is often used as a screening tool for detecting intracranial stenosis. Since TCD results may vary among laboratories and sonographers, it requires validation against an imaging modality. We evaluated diagnostic accuracy of TCD performed in our laboratory against time-of-flight (TOF) magnetic resonance angiography (MRA) in detecting intracranial stenosis in patients with acute cerebral ischemia. METHODS: Consecutive patients with acute (<24 hours) cerebral ischemia and intracranial arterial stenosis on MRA underwent both TCD and MRA within 5 hours of each other. TCD was performed by credentialed neurosonologists according to standardized protocol. An independent neuroradiologist interpreting MRA was blinded to TCD findings. We evaluated TCD peak systolic velocities (PSV) in proximal intracranial arteries as predictive of moderate (>50%) and severe (>70%) stenosis on TOF-MRA. RESULTS: One hundred and fifty patients (74% males; mean age 53 years) underwent neurovascular evaluation with TCD and MRA. Twenty-two (14.6%) patients were excluded due to absent temporal acoustic windows. Middle cerebral artery TCD PSV values of >140 cm/s and >180 cm/s were found to predict a >50% and >70% focal stenosis, respectively. Optimal cut-off PSV values for other major proximal intracranial arteries were also established. CONCLUSIONS: TCD performed in our laboratory shows satisfactory agreement with TOF-MRA in diagnosis and grading of proximal intracranial stenosis in patients with acute cerebral ischemia. PMID- 20002972 TI - Effect of different cements on the biomechanical behavior of teeth restored with cast dowel-and-cores-in vitro and FEA analysis. AB - PURPOSE: To test the hypothesis that the type of cement used for fixation of cast dowel-and-cores might influence fracture resistance, fracture mode, and stress distribution of single-rooted teeth restored with this class of metallic dowels. MATERIALS AND METHODS: The coronal portion was removed from 40 bovine incisors, leaving a 15 mm root. After endodontic treatment and standardized root canal relief at 10 mm, specimens were embedded in polystyrene resin, and the periodontal ligament was simulated with polyether impression material. The specimens were randomly divided into four groups (n = 10), and restored with Cu Al cast dowel-and-cores cemented with one of four options: conventional glass ionomer cement (GI); resin-modified glass ionomer cement (GR); dual-cure resin cement (RC); or zinc-phosphate cement (ZP). Sequentially, fracture resistance of the specimens was tested with a tangential load at a 135 degrees angle with a 0.5 mm/min crosshead speed. Data were analyzed using one-way analysis of variance (ANOVA) and the Fisher test. Two-dimensional finite element analysis (2D-FEA) was then performed with representative models of each group simulating a 100 microm cement layer. Results were analyzed based on von Mises stress distribution criteria. RESULTS: The mean fracture resistance values were (in N): RC, 838.2 +/- 135.9; GI, 772.4 +/- 169.8; GR, 613.4 +/- 157.5; ZP, 643.6 +/- 106.7. FEA revealed that RC and GR presented lower stress values than ZP and GI. The higher stress concentration was coincident with more catastrophic failures, and consequently, with lower fracture resistance values. CONCLUSIONS: The type of cement influenced fracture resistance, failure mode, and stress distribution on teeth restored with cast dowel-and-cores. PMID- 20002973 TI - Effect of 180 days of water storage on the transverse strength of acetal resin denture base material. AB - PURPOSE: Acetal resin has been used as an alternative denture base and clasp material since 1986. The manufacturers claim that acetal resin has superior physical properties when compared to conventional denture base acrylic resins. Limited information is available about transverse strengths of acetal resin. The purpose of this investigation was to compare transverse strengths of pink and white acetal resins to transverse strengths of conventional heat-polymerized polymethylmethacrylate (PMMA) resin in increasing durations of water storage. MATERIALS AND METHODS: A transverse strength test was performed in accordance with International Standards Organization (ISO) specification No 1567. Twenty 65 x 10 x 2.5 mm(3) specimens of each resin were prepared; five specimens of each resin group were subjected to three-point bending test after 50 hours, 30 days, 60 days, and 180 days of water storage in distilled water at 37 degrees C. Experimental groups' transverse strengths were compared by three-way ANOVA and Duncan's multiple range tests. RESULTS: Transverse strength of PMMA denture base material was found to be in accordance with the requirements of ISO specification No 1567. Transverse strengths of white and pink acetal resin could not be calculated in this study, as white and pink acetal resin specimens did not break at the maximum applied force in the three-point bending test. Flexural strength of acetal resin was found to be within the ISO specification limits. As the water storage time increased, the deflection values of PMMA showed no significant difference (p > 0.05). Both the white and pink acetal resin showed significant increase in deflection as the water storage time was increased from 50 hours to 180 days (p < 0.05). CONCLUSION: The results of this study indicated that transverse strength values of PMMA were within the ISO specification limit. Water storage time (50 hours, 30, 60, and 180 days) had no statistically significant effect on the transverse strength and deflection of PMMA. Acetal resin suffered from permanent deformation, but did not break in the three-point bending test. Acetal resin showed significant increase in deflection as the water storage time was increased from 50 hours to 180 days. All materials tested demonstrated deflection values in compliance with ISO specification No 1567. PMID- 20002974 TI - The history of articulators: the "Articulator Wars" phenomenon with some circumstances leading up to it. AB - At the dawn of the 20th century, all was not well with the practice of "plate prostheses." Removable prosthodontics had been degrading for several decades and was now generally in low esteem, even though there had been many significant advances. W. E. Walker had introduced adjustable condylar guides, George Snow, the facebow, and Carl Christensen, a method for clinically measuring the condylar inclines. Nevertheless, the average practicing dentist was still using simple hinge articulators and was apathetic to the deplorable state of the artificial teeth available; however, this was all going to change dramatically when two dentists, Alfred Gysi and J. Leon Williams, working together between 1910 and 1914, presented to the profession the "Trubyte Artificial Tooth System" that embodied both a typal system for selecting anterior teeth and new posterior occlusal carvings that made possible, for the first time, the articulation of artificial teeth. This incited many of prosthetic dentistry's elite to introduce their own theories of mandibular movement and the articulators that they designed to reflect those theories. The intense debates that ensued, both in the meeting halls and in the literature, were numerous and lasted for decades. At the time, the "Articulator Wars" had both positive and negative consequences. Today, with many of the "Articulator Wars" issues remaining as part of the practice of dentistry, the "Articulator Wars" can be considered a phenomenon of enlightenment. PMID- 20002975 TI - Survey of currently used materials for fabrication of extraoral maxillofacial prostheses in North America, Europe, Asia, and Australia. AB - PURPOSE: The purpose of this survey was to review the extraoral maxillofacial materials currently used as well as the advantages and disadvantages of the materials in the fabrication of facial prostheses. Results of this survey will enhance scientific knowledge, generate research study ideas, and possibly lead to production of alternative or new maxillofacial materials. MATERIAL AND METHODS: A 47-question survey was delivered via e-mail to all members (combined total of 260 members) of the American Anaplastology Association (AAA) and American Academy of Maxillofacial Prosthetics (AAMP) for evaluation of personal preference involving maxillofacial prosthetic materials (intrinsic/extrinsic silicone elastomers and pigments/colorants used, polymerization/curing process, advantages and disadvantages of the most often used materials, most important characteristic of material/technique used). RESULTS: The views of 43 (16%) respondents indicated that the majority surveyed were using room temperature-vulcanized (RTV) silicone products. Silicone pigments for intrinsic and silicone pastes for extrinsic coloring were favored over artist's oil colors and dry earth pigments. The polymerization process and/or curing times and temperatures for the same silicone material varied between users. The top five advantages of most often used materials were good esthetics, ease of coloring, easy manipulation, thin margins possible, and adhesive compatibility. The top five disadvantages were discoloration over time, technique-sensitivity, lack of repairability, extrinsic colors peel/fade, and lack of longevity. Nontoxic/nonallergenic materials with high edge strength and color stability were the most important features when choosing a maxillofacial prosthetic material/technique. CONCLUSIONS: The responses to this survey indicate that the majority of AAA and AAMP members are using or have used a variety of RTV silicones, pigments, and colorants in the quest to provide the best possible facial prosthetic service. Further research is needed to further refine and improve extraoral maxillofacial materials/techniques based on the results of this study. PMID- 20002976 TI - Advanced education in prosthodontics: residents' perspectives on their current training and future goals. AB - PURPOSE: The purposes of this study were to identify current prosthodontic residents' demographics and to document prosthodontic residents' perspectives on their clinical training and future goals. MATERIALS AND METHODS: A 52-item survey was created and distributed to prosthodontic residents in the United States on February 8, 2007. The data collected were analyzed; the means and standard deviations were calculated and ranked. Statistical analysis was conducted using Chi-square and Mann-Whitney analysis (p = 0.05). RESULTS: A 43% response rate was achieved, representing approximately 48% of the total population of prosthodontic residents in the United States. The majority of residents ranked clinical education as the most important factor in selecting their programs, were satisfied with their training, and planned to pursue the certification of the American Board of Prosthodontics. When asked how often they planned to work, 4 days a week was the most common answer. CONCLUSION: This is the first report identifying current prosthodontic residents' demographics and their perspectives on their clinical training and future goals. Several trends were identified, indicating a bright future for the specialty. By knowing the students' perceptions regarding their training and future goals, the American College of Prosthodontists and/or program directors will be able to use this information to improve residency programs and the specialty. PMID- 20002977 TI - Effects of clinical experience and explorer type on judged crown margin acceptability. AB - PURPOSE: This study was undertaken to simultaneously compare instrumentation type and operator characteristics in judgments of clinical acceptability of crowns exhibiting a controlled range of marginal gaps. The research was conducted in a laboratory setting and generalizability analysis was used as a statistical technique to identify the sources contributing to variation in the judgment outcome. MATERIALS AND METHODS: A crown was seated on an ivorine tooth in a device that permitted continuous adjustment in intervals of 25 mum to produce known marginal gaps ranging from zero to 250 microm. Forty-nine students and six faculty members used five types of explorers each to determine, by tactile examination, the point on the controlled increasing marginal gap where they would no longer regard the gap as clinically acceptable. RESULTS: There were no differences across type of explorer. Operators with clinical experience had a threshold that rejected crowns at a smaller gap than did those operators without clinical experience (p= 0.007). Faculty members maintained a higher individual degree of consistency in their personal judgments than did students (p= 0.02); however, the inter-operator consistency was significantly lower for faculty members than for students (p < 0.05). CONCLUSIONS: Differences among operators in a simulation of the decision regarding gaps in crowns accounted for 63% of the variance; type of explorer used in assisting this decision accounted for about half as much variance. Faculty members making such judgments exhibited high intra operator consistency but significantly lower inter-operator consistency than did students. The study suggests that the internal standards dentists use for clinical decision making deserves further study as they may be as significant as the equipment used. PMID- 20002978 TI - Color stability of denture acrylic resins and a soft lining material against tea, coffee, and nicotine. AB - PURPOSE: The purpose of this study was to investigate the effect of four solutions [saliva (control group), saliva+tea, saliva+coffee, saliva+nicotine] on the color of different denture base acrylic resins (heat-polymerized, injection molded, autopolymerized) and a soft denture liner. MATERIALS AND METHODS: Twenty specimens from each type of test material were prepared (2.5 mm diameter, 2 mm thickness). Five specimens from each test material (heat-polymerized, chemically polymerized, injection-molded acrylic resin, soft denture reliner) were stored in each solution in 37 degrees C in a dark environment. Colorimetric measurements were done on the 1st, 7th, and 30th days. Color differences among specimens immersed in saliva (control group), and staining solutions were evaluated over time. Data were statistically analyzed with one-way analysis of variance (ANOVA) (alpha= 0.05). ANOVA was followed by Tukey test to find which groups differed from each other. RESULTS: Significant color shifts occurred in heat-polymerized and injection-molded acrylic resins in coffee and in soft liner in nicotine over time (p < 0.05) (DeltaE > 1). The color shift of soft liner in nicotine was significantly different than that of the remainder of the test materials in nicotine (p < 0.05). The color shift magnitudes of each test material in coffee and tea were not significantly different when compared among the test material groups (p > 0.05). CONCLUSIONS: The effect of staining solutions on the color of each test material in each session was perceivable by the human eye (DeltaE > 1); however, the color shifts of all test materials were clinically acceptable (DeltaE < 3.7) except for soft liner in nicotine, which was not clinically acceptable over time. Therefore, minimizing drinking of such beverages and use of tobacco, particularly when soft liner is applied, may be advantageous for denture wearers for long-term color stability. PMID- 20002979 TI - Bond strength of resin cements to Co-Cr and Ni-Cr metal alloys using adhesive primers. AB - PURPOSE: The aim of this study was to evaluate the effectiveness of adhesive primers (APs) applied to Co-Cr and Ni-Cr metal alloys on the bond strength of resin cements to alloys. MATERIALS AND METHODS: Eight cementing systems were evaluated, consisting of four resin cements (Bistite II DC, LinkMax, Panavia F 2.0, RelyX Unicem) with or without their respective APs (Metaltite, Metal Primer II, Alloy Primer, Ceramic Primer). The two types of dental alloys (Co-Cr, Ni-Cr) were cast in plate specimens (10 x 5 x 1 mm(3)) from resin patterns. After casting, the plates were sandblasted with aluminum oxide (100 microm) and randomly divided into eight groups (n = 6). Each surface to be bonded was treated with one of eight cementing systems. Three resin cement cylinders (0.5 mm high, 0.75 mm diameter) were built on each bonded metal alloy surface, using a Tygon tubing mold. After water storage for 24 hours, specimens were subjected to micro shear testing. Data were statistically analyzed by two-way ANOVA and Tukey's studentized range test. RESULTS: The application of Metal Primer II resulted in a significantly higher bond strength for LinkMax resin cement when applied in both metal alloys. In general, the cementing systems had higher bond strengths in Co Cr alloy than in Ni-Cr. CONCLUSIONS: The use of AP between alloy metal surfaces and resin cements did not increase the bond strength for most cementing systems evaluated. PMID- 20002980 TI - Clinicopathological and immunohistochemical features of oral spindle cell carcinoma. AB - BACKGROUND: Oral spindle cell carcinoma (SpCC) is a rare variant of oral squamous cell carcinoma (SCC). The aims of this study were to compare the clinicopathologic and immunohistochemical features of oral SpCC with conventional oral SCC. METHODS: Five cases of oral SpCC and 10 cases of oral SCC (five well differentiated and five poorly differentiated) were evaluated through conventional hematoxylin and eosin staining and immunohistochemical reactions to cytokeratins (CK), vimentin, desmin, smooth muscle actin, muscle-specific actin, S-100 protein, epithelial membrane antigen (EMA), p53, and ki-67. RESULTS: Oral SpCC showed predilection for males on their sixth decade of life, presenting clinically as painful infiltrative ulcers or ulcerated exophytic polypoid masses, preferably located on the alveolar mucosa. Mesenchymal markers were expressed in the spindle cell but not in the carcinomatous component of SpCC, and it was negative in all SCC. CKs AE1/AE3, 6, 14, and EMA were positive on both carcinomatous and spindle cell components of most SpCCs. These tumors also presented higher p53 and ki-67 expression and no CK 1 expression in contrast to well-differentiated SCC. CONCLUSION: Oral SpCC presented a different clinical profile than conventional SCC and histopathologic features and p53 and ki-67 expression closer to poorly differentiated SCC. Besides mesenchymal markers, CK AE1/AE3, 6, 14, and EMA expression on spindle cells may be useful as an adjunct on microscopical differential diagnosis of SpCC. PMID- 20002981 TI - Mode of HIV transmission associated with risk of oral lesions in HIV-infected subjects in Thailand. AB - BACKGROUND: The aim of this study was to determine if route of human immunodeficiency virus (HIV) transmission is associated with the risk of oral lesions in HIV-infected subjects in Thailand. METHODS: A cross-sectional study was performed in 186 HIV-infected heterosexuals (aged 21-65 years, mean 32 years), and 82 HIV-infected intravenous drug users (IVDUs) (aged 16-50 years, mean 30 years). The following information was recorded: route of HIV transmission, total lymphocyte cell counts, weight, smoking habit, alcohol consumption, medications, presence of denture, plaque index, and presence of oral lesions. The association between mode of HIV transmission and the risk of oral lesions among the subjects was determined by multiple logistic regression analysis. RESULTS: Oral lesions were found in 138 HIV-infected heterosexuals (75%) and in 37 HIV-infected IVDUs (46%). Oral candidiasis (OC) was the most common lesion among both groups (44% vs. 28%), followed by hairy leukoplakia (HL) (33% vs. 10%). Multiple logistic regression analysis showed a significant association between mode of HIV transmission and the risk of oral lesions after controlling for the total lymphocyte cell counts and other confounding factors [OR 3.1; 95% CI 1.5-6.4; P = 0.002]. OC was significantly associated with heterosexual route of HIV transmission [OR 2.4; 95% CI 1.2-4.7; P = 0.014]. Similar association was also observed with HL [OR 3.7; 95% CI 1.5-9.1; P = 0.004]. CONCLUSIONS: Mode of HIV transmission is associated with the risk of oral lesions in HIV-infected subjects in Thailand. Further studies should be performed to determine if the risk of oral lesions is associated with differences in HIV subtypes. PMID- 20002982 TI - Evaluation of correlation of serum lipid profile in patients with oral cancer and precancer and its association with tobacco abuse. AB - BACKGROUND: Cholesterol at either higher/lower level can be troublesome. Health issues related to higher than normal levels have received much public attention because of their relationship to incidence of heart disease, whereas implications of decreased cholesterol levels remain unclear. Present study tried to evaluate and correlate the decreased cholesterol levels in Oral cancer, Oral precancer and in tobacco abuse. METHODS: Total Cholesterol (TC), High Density Lipoproteins (HDL), Very Low Density Lipoproteins (VLDL), Low Density Lipoproteins (LDL) and Triglyceride (Tri) were estimated in 210 subjects. Out of these 210 subjects, 70 subjects were histopathologically confirmed Oral Cancer, 70 subjects were histopathologically confirmed Oral precancer (OPC) and 70, age and sex matched, healthy subjects who are not having Oral Cancer, Oral precancer and who had no history of any major illness in the past. These groups were subdivided into: Subjects with No Habit of Tobacco (NHT) and Subjects With Habit of Tobacco (WHT). RESULTS: There was significant decrease in TC, HDL, VLDL, and triglyceride in Oral Cancer group; and significant decrease in TC, and HDL in Oral precancer group as compared to Control. Mean serum lipid profile levels were not significantly different in subjects between NHT and WHT. CONCLUSIONS: There is an inverse relationship between serum lipid profile and Oral Cancer and Oral precancer. There was no overall significant correlation of serum lipid profile with tobacco abuse. PMID- 20002983 TI - Face transplantation. PMID- 20002984 TI - End-of-life decision making - where have all the surgeons gone? PMID- 20002985 TI - Fatwas and surgery: how and why a fatwa may inform a Muslim patient's surgical options. PMID- 20002986 TI - Developing academic surgeons: the focus of a new course. PMID- 20002988 TI - Surgical history of ancient China: part 1. AB - Although surgery was an accepted and quite proficient craft very early on in Chinese history, it has deteriorated through the ages. Despite the fact that anaesthetic agents in major surgery were employed during the third century, Chinese surgery is conspicuous by its stagnation. Reverence for the dead, filial piety, abhorrence of shedding blood and other conservative attitudes make it impossible for any accurate knowledge of the human anatomy and physiology, without which surgery cannot progress. This article surveys some highlights in the history of surgery in ancient China and examines the factors responsible for its decline. The second concluding part deals with orthopaedics. PMID- 20002989 TI - Surgical decision making. AB - Decision making is an integral part of surgical practice and is a skill that is as important as the ability to operate. Sound decision making is the result of acquiring 'surgical wisdom', which is achieved through learning, experience and reflection. This paper reviews the processes of surgical decision making with respect to choosing the correct procedure and deciding when to operate, and how operative decisions are made, together with the factors that influence our surgical decisions. PMID- 20002990 TI - Facial transplantation as an option in reconstructive surgery: no mountains too high? AB - BACKGROUND: Human facial allotransplantation (FA) is a testament to the impressive progress, which was and is still happening in the field of transplantation medicine and reconstructive surgery. Like every pioneering treatment option, FA faces both the clinicians' and public opinion with a huge amount of medical and psychosocial dilemmas, which, in order to introduce FA as a scientifically and socially accepted procedure into clinical practice, need to be discussed, answered and solved. METHODS: This review is based on a detailed literature research in all relevant databases (MEDLINE, PubMed, Cochrane Library, specialist textbooks), however, recent papers (published between 2006 and 2008) were given highest priority for inclusion. RESULTS: In this review, two main limitations associated with facial tissue allotransplantation, that is, shortage of donations and life-long need for immunosuppressive treatment, are discussed and presented in both psychosocial and medical terms. CONCLUSION: Although both of these limitations potentially could successfully inhibit the transformation of FA from an experimental therapy to the treatment of choice for patients with severe functional facial impairment, recent literature suggest that FA will find a meaningful place in facial reconstructive surgery. PMID- 20002991 TI - Fijian seasonal scourge of mango tree falls. AB - BACKGROUND: Mango tree falls are a frequent presentation at any health facility in the South Pacific. This study aims to identify (i) the number of admissions because of falls from mango trees; (ii) epidemiology; (iii) seasonal trend; (iv) injury profile; and (v) hospital care provided. METHODS: Retrospective case review on all mango tree falls related injuries resulting in admissions at the Lautoka Hospital, Fiji during a 1-year period (2007). Patient records were analysed to identify specific injury patterns such as upper/lower limb fractures, spinal cord injury and head injury, caused by mango tree falls. RESULTS: Thirty nine cases were identified. Eighty-two percent (n = 32) of the falls occurred in the mango season (June-November). Seventy-two percent (n = 28) of the patients were males and 28% (n = 11) were females. Seventy-six percent were aged 5-13 and only 21% were adults. Also, 77% (n = 30) of the patients were ethnic Fijians and 23% (n = 9) were Fijian-Indians. Sixty-four percent (n = 25) had closed fractures and 17% (n = 7) had open fractures. Fifty-six percent (n = 22) of the fractures were of the fore arm. There were two cases of spinal cord injury, four cases of head injury, one ICU admission and one death. Average hospital stay was 7.56 days. CONCLUSION: All these injuries were recreational and the majority in the urban setting. They were all avoidable. PMID- 20002992 TI - Famous discoveries by medical students. AB - Heparin, insulin, the sinoatrial node and ether anaesthesia are just some of the major discoveries made by medical students, all of which have had a huge impact on the practice of surgery. This paper explores the breadth and depth of some of these talented individuals and their extraordinary contributions to medicine. While some continued to enjoy brilliant careers, others became embroiled in damaging disputes about priority or were overtaken by illness or personal problems. Regardless of their fate, their contributions are a potent reminder of what can be achieved by gifted and determined medical students undertaking a period of basic scientific or clinical research. PMID- 20002993 TI - Nothing to sneeze at! A study into intra-operative contamination. AB - PURPOSE: We performed a prospective study of sneezes from orthopaedic registrars to assess the potential for intra-operative contamination from a masked surgeon, and to determine if head position can alter the potential for contamination. TYPE OF STUDY: Prospective controlled trial. METHODS: Four orthopaedic registrars from the Gold Coast Hospital each inhaled pepper to precipitate a sneeze. Cultures were taken with and without standard Smith & Nephew surgical masks, in positions directly in front and to the sides of a masked registrar. The process was repeated three times for each registrar. A control plate was left exposed to the atmosphere. A control plate and sneeze plate was cultured on blood agar. Three masks were subsequently swabbed to exclude contamination from the masks. RESULTS: 2/24 (8.33%) of the side with mask cultures returned significant bacterial growths. Also, 1/12 (8.33%) of the front with mask cultures returned significant growths. In addition, 9/12 (75%) of the direct sneezes resulted in significant bacterial counts. The control plate failed to return any growth. Subsequent culture of the exterior of three surgical masks failed to yield significant growth. There was a statistically significant odds ratio of 0.03 comparing the front and side group with the unmasked direct group. There was no statistically significant difference comparing front and side sneeze growth. CONCLUSIONS: While the use of surgical face masks significantly reduces bacterial counts following a sneeze, it fails to eliminate the potential for surgical field contamination completely. The fact that significant bacterial counts can be returned from direct culture through a mask and to the sides of a mask suggests that head position is irrelevant and contamination is possible in any direction. It is, therefore, suggested that, if possible, a sneezing surgeon distances himself/herself as much as possible from the sterile field. We also recommend following a sneeze; surgeons should re-gown and glove, given the risk of contamination of the sterile field. LEVEL OF EVIDENCE: Level II. PMID- 20002994 TI - Evaluating anatomical research in surgery: a prospective comparison of cadaveric and living anatomical studies of the abdominal wall. AB - BACKGROUND: Cadaveric research has widely influenced our understanding of clinical anatomy. However, while many soft-tissue structures remain quiescent after death, other tissues, such as viscera, undergo structural and functional changes that may influence their use in predicting living anatomy. In particular, our understanding of vascular anatomy has been based upon cadaveric studies, in which vascular tone and flow do not match the living situation. METHODS: An angiographic analysis of the abdominal wall vasculature was performed using plain film and computed tomography angiography in 60 cadaveric hemi-abdominal walls (from 31 cadavers) and 140 living hemi-abdominal walls (in 70 patients). The deep inferior epigastric artery (DIEA) and all of its perforating branches larger than 0.5 mm were analysed for number, calibre and location. RESULTS: Both large, named vessels and small calibre vessels show marked differences between living anatomy and cadaveric specimens. The DIEA was of larger diameter (4.2 mm versus 3.1 mm, P < 0.01) and had more detectable branches in the cadaveric specimens. Perforators were of greater calibre (diameter 1.5 mm versus 0.8 mm, P < 0.01) and were more plentiful (16 versus 6, P < 0.01) in cadaveric specimens. However, the location of individual vessels was similar. CONCLUSIONS: Cadaveric anatomy displays marked differences to in vivo anatomy, with the absence of living vascular dynamics affecting vessel diameters in cadaveric specimens. Blood vessels are of greater measurable calibre in cadaveric specimens than in the living. Consequently, cadaveric anatomy should be interpreted with consideration of post-mortem changes, while living anatomical studies, particularly with the use of imaging technologies, should be embraced in anatomical research. PMID- 20002995 TI - William Marsden, a pioneer in social medicine. PMID- 20002996 TI - John Brown's book. PMID- 20002997 TI - Sir William Mackenzie: sympathetic ophthalmia and glaucoma before ophthalmoscopy. AB - One of the practitioners of probably the oldest surgical specialty, ophthalmic, was the eminent Scottish ophthalmologist, Sir William Mackenzie. Educated in Edinburgh, he moved to Glasgow, and described and named sympathetic ophthalmia before the time of the ophthalmoscope, well defining his powers of observation and deduction. Founding the Glasgow Eye Infirmary, his 'Practical Treatise on Diseases of the Eye' appeared in English in four editions (1850-1884) and in French and German. In this also appears the first full and clear account of glaucoma. Both he and the illustrator of his book, Wharton Jones, moved to Glasgow because of rather indefinite connections with Robert Knox, the anatomist, who was allegedly helped by the bodysnatchers, Burke and Hare. Mackenzie and his book were highly regarded before the revolutionary ophthalmoscope. He was knighted and appointed Surgeon Oculist to the Queen in Scotland. PMID- 20002998 TI - From Lanfranc to Sunderland: the surgery of peripheral nerve injuries. PMID- 20002999 TI - Signal naval achievements of James Lind (1747), James Cook (1770) and Owen Stanley (1847). PMID- 20003000 TI - Art of preserving health: studies on the medical supervision of physical exercise. AB - To the ancient Greeks, physical exercise was an essential part of life, especially during adolescence and young adulthood. Long after the end of the Classical Greek era, Roman conquest brought a shift towards martial training, increased professionalism in athletic competition and a weak strand of restorative gymnastics kept barely intact by the likes of Galen. While the crux of these teachings was the use of exercise, among other things, to promote and maintain health, the emphasis began to shift to concerns about the health of athletes and the medical problems brought about by exercise. Fashions in athletic training began to change in the mid-nineteenth century, but the mystique associated with athletic training pervaded much of the thinking and still persists today where, in this modern scientific period of exercise and health, physiologists, physical educators and physicians have become involved in seeking to apply the scientific method to what has become known as exercise science. The modern concept of sports medicine tends to emphasize the training and welfare of the elite athlete. PMID- 20003001 TI - Xanthogranulomatous cholecystitis masquerading as malignancy with liver metastasis. PMID- 20003002 TI - Multinodular goitre complicated by abscess due to E. coli. PMID- 20003003 TI - Large retroperitoneal calcified hydatid cyst. PMID- 20003007 TI - Water use in the surgical scrub: surgeons can reduce their environmental footprint. PMID- 20003008 TI - Water use in the surgical scrub: isn't it time to stop using water at all? PMID- 20003009 TI - Prognostic endovascular scoring is useful for the assessment of patients with additional pathology. PMID- 20003010 TI - Rare complication following Longo operation: giant rectal haematoma. Case report. PMID- 20003011 TI - Mass not suitable for liposuction. PMID- 20003012 TI - Interdisciplinary resection of a giant retroperitoneal liposarcoma of 25 kg. PMID- 20003013 TI - Spontaneous resolution (and other rarities) of gallstone ileus. PMID- 20003014 TI - Bilateral recurrent laryngeal nerve palsy after Ivor Lewis oesophagectomy. PMID- 20003015 TI - A rare case of small bowel intussusception through a loop ileostomy. PMID- 20003016 TI - Bier's block and regional anaesthesia techniques in hand surgery. PMID- 20003017 TI - Continuing medical education program in echocardiography. AB - Article Title: Association between Left Atrium Enlargement and Intradialytic Hypotension: Role of Diastolic Dysfunction in the Hemodynamic Complications during Hemodialysis. PMID- 20003018 TI - Differential left atrial remodeling in LV diastolic dysfunction and mitral regurgitation. AB - OBJECTIVE: Chronic pressure and volume overload may cause different type of left atrial (LA) remodeling in left atrial enlargement (LAE) leading to different cardiovascular outcomes. These two different patterns of LA remodeling can be discriminated by LA eccentricity index (LAEi). The goal of our study was to evaluate an association between LAEi, LV diastolic dysfunction (LVDD), and mitral regurgitation (MR). METHOD: LAEi was calculated from 3D of left atrium (LA): anteroposterior (D1), superoinferior (D2), mediolateral (D3), and LAEi = D2x2/ (D1+D3). LAE was described as elongated left atrium (EA) if LAEi > or = 1.27, and spherical left atrium (SA) if LAEi < 1.27. RESULTS: A group of 501 patients (10.4%) with LAE were categorized into two subgroups; 232 (46.3%) with EA and 269 (53.7%) with SA based on LAEi. Among 108 patients (21.6%) with LVDD without MR, 102 had EA and only 6 had SA (P < 0.0001). The cohort of 155(30.1%) patients with MR without LVDD, only 8 had EA and143 had SA (P < 0.0001). Of the total group of 501 patients, 59 had persistent AF and in this subgroup only 10 patients had EA and 49 patients had SA (P < 0.0001). There was a statistically significant difference for AF rate between EA-patients with LVDD without MR and SA-patients with MR without LVDD (P < 0.001). CONCLUSIONS: LVDD may contribute to pressure overload LA remodeling in a way quite different from volume overload LA remodeling by MR. LAEi can discriminate two different LA morphologies with significantly different potential outcomes. It may identify patients associated with a higher rate of AF with a higher cardiovascular risk. PMID- 20003019 TI - Safety and feasibility of real time adenosine myocardial contrast echocardiography with emphasis on induction of arrhythmias: a study in healthy volunteers and patients with stable coronary artery disease. AB - INTRODUCTION: Some studies reported an increased incidence of premature ventricular complexes (PVCs) during triggered myocardial contrast echocardiography (MCE) using high-intensity ultrasound destruction. Whether PVCs are also induced by real time MCE using low emission power, is unknown. The aim of the study was to assess the occurrence of arrhythmias during real time adenosine MCE in healthy volunteers and patients with stable coronary artery disease (CAD). METHODS: Fifty healthy volunteers and 26 patients with stable CAD underwent real time MCE using Sonovue and power pulse inversion (ATL 5000) at rest and during adenosine stress. The occurrence of premature atrial complexes (PAC) and PVCs was analyzed before and during MCE using ECG-tracings from videotapes. RESULTS: In healthy subjects, the occurrence of PVCs at baseline (0.04 +/- 0.23 PVCs/min) was similar at rest (0.04 +/- 0.23 PVCs/min, P = NS), and adenosine stress (0.03 +/- 0.14, P = NS). In CAD patients, the occurrence of PVCs at baseline was 0.30 +/- 0.76 PVC/min, compared to 0.29 +/- 0.74 at rest (P = NS), and 0.34 +/- 0.74 during adenosine stress (P = NS). The number of subjects demonstrating PVCs did not increase during MCE. The occurrence of PACs during MCE was not increased compared to baseline. CONCLUSION: Real time MCE using low emission power does not increase the occurrence of premature complexes in healthy volunteers or CAD patients. PMID- 20003020 TI - Can 3D ultrasound and doppler angiography of great arteries be included in second trimester ecocardiographic examination? A prospective study on low-risk pregnancy population. AB - The introduction of new applications such as the spatiotemporal image correlation (STIC) and the tomographic ultrasound imaging (TUI) has greatly enhanced the study of cardiac and vessels anatomy. The aim of our study was to assess the feasibility of including real-time volumetric 3D reconstruction of heart anatomy and Doppler angiography of great vessels during routine second trimester pregnancy scan, in a low-risk population. Visualization of the pulmonary veins, ductus venosus and inferior vena cava was significantly superior with 3D ultrasound. Therefore, STIC makes it possible to store heart beats for a mean time of 10 s, to replay acquired volumes in slow motion, to study the heart atrium along three axes and retrace volumetric image storage inversely from the apex to the base of the heart, making reconstruction of the four-chamber view and great artery outflow tract much easier and faster. Furthermore, the use of digital communication allow operators to store and send volume of images that can be postprocessed by external consultant specialists worldwide. Although 3D echocardiography is less time-consuming and independent of fetal positioning in utero when compared with 2D it may be premature to include 3D and 3D Doppler angiography among routine examination. However, the study of the outflow tracts by 2D echocardiography should be implemented in order to improve the diagnostic accuracy of CHDs. At present, only "targeted" cases in which a routine 2D scan has raised diagnostic doubts should undergo a second level sonography for a thorough 2D-3D examination of cardiac and vessels anatomy. PMID- 20003021 TI - Flow-mediated dilatation: a review of techniques and applications. AB - Cardiovascular disease is one of the leading causes of death in industrialized nations worldwide. Traditional assessment of cardiovascular disease often involves invasive procedures (e.g., coronary angiography) and also provides little functional information of arterial reactivity. The emergence of the assessment of "subclinical" atherosclerosis and the role of arterial stiffness in both cardiovascular prognosis and cardiovascular risk have become more widely accepted and used in modern clinical practice and research. Flow-mediated dilatation can be used to directly assess endothelial function in healthy and diseased populations. This review aims to provide the reader with an insight into the technique and clinical applications of this recently new tool for the assessment of cardiovascular disease and risk. PMID- 20003022 TI - The utility of live three-dimensional echocardiography in the surgical treatment of bilateral coronary fistulas. AB - In patients with coronary and pulmonary arterial fistulas, the drainage sites may be difficult to identify if there are multiple connections by standard 2D echocardiography techniques. However, real time 3D TEE was used intraoperatively in our case to facilitate site identification, the number of fistulous connections, and documentation of the fistula ligation. PMID- 20003023 TI - Two-dimensional echocardiographic findings of pulmonary valve cyst secondary to pulmonary valvuloplasty. AB - Real time two-dimensional transthoracic and transesophageal echocardiography demonstrated a mobile echolucent mass attached to the pulmonary valve in a 25 year-old adult, 20 years following balloon pulmonary valvuloplasty. The mass was surgically excised and pathology showed it to be a cyst. PMID- 20003024 TI - Images in echocardiography: a freely floating saphenous vein graft. PMID- 20003025 TI - Fatal coronary embolism in infective endocarditis associated with chronic rheumatic heart disease: an unexpected transesophageal echocardiographic documentation. PMID- 20003026 TI - Ruptured sinus of valsalva aneurysm in a Turner's syndrome patient. PMID- 20003027 TI - Complement C3 in Bernese Mountain dogs. AB - BACKGROUND: Previous research suggests that low serum concentrations of the third component of complement (C3) are associated with both the susceptibility to infectious agents such as Borrelia burgdorferi and the development of glomerular disease. We hypothesized that low levels of C3 are associated with the coincident occurrence of B. burgdorferi infection and glomerulonephritis in Bernese Mountain dogs. OBJECTIVES: The aims of this study were to evaluate the serum concentration of C3 in Bernese Mountain dogs with and without antibodies against B. burgdorferi and to compare this concentration with that of healthy control dogs. METHODS: Eighty-three clinically healthy Bernese Mountain dogs and 46 control dogs were included. Antibodies against B. burgdorferi were determined using an ELISA with a whole cell sonicate as antigen. Results were confirmed using Western blot. C3 was measured using a single radial immunodiffusion test. Results were reported as the percentage concentration of C3 compared with that in pooled preserved canine serum (100% C3 concentration). RESULTS: Median C3 concentration was 128.5% in Bernese Mountain dogs with antibodies against B. burgdorferi, 133.5% in B. burgdorferi-negative Bernese Mountain dogs, 87.8% in positive control dogs, and 102.2% in negative control dogs. Within Bernese Mountain and control groups, C3 was lower in dogs with antibodies against B. burgdorferi compared with those without. Percentage concentration of C3 was higher in healthy Bernese Mountain dogs compared with control dogs. CONCLUSION: Low C3 concentration is not an explanation for the high prevalence of B. burgdorferi infections and glomerular disease in Bernese Mountain dogs. PMID- 20003028 TI - Evaluation of peripheral blood neutrophil function in tumor-bearing dogs. AB - BACKGROUND: Peripheral blood neutrophils of untreated human cancer patients have been shown to have normal, increased, and decreased phagocytic activity, killing capacity, and/or oxidative burst activities. OBJECTIVES: The objectives of this study were to evaluate oxidative burst and phagocytic activities of peripheral blood neutrophils from tumor-bearing dogs before therapy and compare them with neutrophil function of healthy control dogs. METHODS: Heparinized whole blood was obtained from dogs with high-grade lymphoma (n=23), sarcoma (n=13), or carcinoma (n=11), and healthy control dogs (n=11) for flow cytometric evaluation of oxidative burst and phagocytic activities. Percentage of bursting cells and amount of oxidative burst activity were determined after stimulation with phorbol 12-myristate 13-acetate (PMA) or Escherichia coli. Percentage of phagocytic cells and amount of phagocytic activity were determined after incubation with fluorescent E. coli. RESULTS: Compared with control dogs, dogs with sarcoma (P=.004) and carcinoma (P=.05) had a lower percentage of neutrophils exhibiting oxidative burst activity after stimulation with PMA. Phagocytic activity was significantly lower in dogs with sarcomas compared with control dogs (P<.0001) and dogs with lymphoma (P=.01). CONCLUSIONS: Untreated carcinomas and sarcomas in dogs may suppress the percentage of neutrophils capable of oxidative burst when stimulated by PMA. Furthermore, sarcomas also may suppress the amount of phagocytic activity per neutrophil. Until further studies can be performed, the clinical significance of these findings is unknown. PMID- 20003029 TI - Measurement of platelet aggregation in ovine blood using a new impedance aggregometer. AB - BACKGROUND: Whole blood platelet aggregometry (impedance) is an important method to investigate platelet function disorders. Examination of hemostatic function in sheep is important with respect to their role as an animal model of human disease. OBJECTIVE: The aim of this study was to evaluate and optimize selected methodological aspects (anticoagulant, agonist concentration) of impedance aggregometry in ovine blood using the new Multiplate 5.0 analyzer. METHODS: Blood samples were collected in hirudin anticoagulant from 40 clinically healthy sheep. Samples from selected sheep were collected in citrate, with or without the addition of calcium chloride. The agonists adenosine diphosphate (ADP), collagen, ristocetin, arachidonic acid, and thrombin receptor-activating peptide (TRAP) were added in several concentrations to induce aggregation. RESULTS: Based on maximum aggregation values and internal precision, no significant difference was found between ADP concentrations of 3-10 micromol/L and collagen concentrations of 3-5 microg/mL (P>.05). The lowest interindividual variation of approximately 3 4-fold was seen with 4 and 5 micromol/L ADP and 4 and 5 microg/mL collagen. Ristocetin, arachidonic acid, and TRAP did not induce significant aggregation at any concentration. Aggregation results were significantly lower when measured in citrate- vs hirudin-anticoagulated blood, regardless of the presence of calcium chloride. CONCLUSIONS: Our results indicate that the multiplate impedance aggregometer is suitable for the measurement of platelet aggregation in sheep using optimal agonist concentrations of 4-5 micromol/L ADP and 4-5 microg/mL collagen. Hirudin-anticoagulated blood is the preferred sample material. PMID- 20003030 TI - Donor hypo- and hypernatremia are predictors for increased 1-year mortality after cardiac transplantation. AB - Donor hypernatremia is known to be associated with initial graft dysfunction in liver transplantation. Controversial data exist regarding the impact of sodium dysregulation on patient survival after heart transplantation (HTX). The aim of this study was to investigate the influence of donor sodium levels on survival in a large cohort of heart transplant recipients from the Eurotransplant registry. From 1997 to 2005, all consecutive adult HTX performed in the Eurotransplant region were included into this study (n = 4641 patients). Multivariate analysis was applied to investigate possible clinical predictors for 1-year post transplant survival after cardiac transplantation (donor sodium levels, donor age, donor cause of death, recipient age, primary disease, urgency status, cold ischemia time). In multivariate analysis, recipients receiving a donor heart with serum sodium level lower than 130 mmol/l or higher than 170 mmol/l had a 1.25 fold higher risk for 1-year post-transplant mortality than patients with normal donor sodium ranges (P = 0.007). Other independent risk factors for impaired 1 year survival were recipient age, the indication for transplantation and the urgency status of the recipient. Our study demonstrates that hyponatremia as well as hypernatremia show a strong U-shaped correlation with poor survival after cardiac transplantation. Accurate donor management to avoid electrolyte disorder seems to be crucial for ensuring good quality of donor hearts. PMID- 20003031 TI - Prevalence of anti-HLA antibodies after liver transplantation. PMID- 20003032 TI - Low levels of high-density lipoprotein cholesterol: an independent risk factor for late adverse cardiovascular events in renal transplant recipients. AB - Long-term kidney transplant graft and patient survival is often limited by cardiovascular (CV) disease. Risk factors for CV disease such as diabetes, hypertension and elevated low-density lipoprotein levels are well documented; however, the impact of low levels of high-density lipoprotein (HDL) has not been defined. We performed a retrospective chart review of 324 consecutive renal transplant recipients from 2001 to 2007 to correlate baseline HDL levels with major adverse cardiovascular events (MACEs) defined as a composite of new onset CV illness, cerebral vascular events and peripheral vascular disease. A total of 92 MACEs occurred over a total of 1913 patient years of follow-up. Low HDL cholesterol levels were noted in 58.3% of patients. Compared with those with normal HDL levels, a greater percentage of patients with low HDL levels had post transplant MACEs (20% vs. 60% respectively) and experienced an increased rate of all cause mortality. Sixty-two percent of all MACEs occurred in patients with low HDL levels. In the low HDL group, the odds ratio for experiencing a MACE was 1.92. Therefore, HDL cholesterol may provide an important new therapeutic target to prevent vascular morbidity and mortality following renal transplantation. PMID- 20003033 TI - Association between birth outcomes and aflatoxin B1 biomarker blood levels in pregnant women in Kumasi, Ghana. AB - OBJECTIVE: To investigate the association between birth outcomes and blood levels of aflatoxin B(1) (AFB1)-lysine adduct in pregnant women in Kumasi, Ghana. METHOD: A cross-sectional study of 785 pregnant women attending antenatal clinic was conducted. Aflatoxin B(1) (AFB(1))-lysine adduct levels were determined by high performance liquid chromatography (HPLC) on blood taken after delivery. The birth outcomes considered were small for gestation age, low birthweight, preterm delivery and stillbirth. Participants were divided into quartiles based on the distribution of aflatoxin B(1)-lysine adducts in pg/mg albumin ('low': 2.67 to 4.97 to 11.34). Statistical analysis involved models that included socio-demographic variables and other potential confounders. RESULTS: The average AFB(1)-lysine adduct level in maternal serum was 10.9 +/- 19.00 pg/mg albumin (range = 0.44-268.73 pg/mg). After adjusting for socio-demographic variables and potential confounding factors, participants in the highest AFB(1)-lysine quartile with 'very high' AFB(1)-lysine level (>11.34 pg/mg) were more likely to have low birthweight babies (OR, 2.09; 95% CI, 1.19-3.68), and showed a trend of increasing risk for low birthweight (P(trend) = 0.007) compared to participants in the lowest quartile. CONCLUSION: This study adds to the growing body of evidence that aflatoxins may increase the risk of adverse birth outcomes. The findings have implications for targeted nutritional education of pregnant women in areas with high levels of aflatoxin contamination of foods. PMID- 20003034 TI - Mortality after failure of antiretroviral therapy in sub-Saharan Africa. AB - OBJECTIVE: To assess the outcome of patients who experienced treatment failure with antiretrovirals in sub-Saharan Africa. METHODS: Analysis of 11 antiretroviral therapy (ART) programmes in sub-Saharan Africa. World Health Organization (WHO) criteria were used to define treatment failure. All ART-naive patients aged >or=16 who started with a non-nucleoside reverse transcriptase inhibitor (NNRTI)-based regimen and had at least 6 months of follow-up were eligible. For each patient who switched to a second-line regimen, 10 matched patients who remained on a non-failing first-line regimen were selected. Time was measured from the time of switching, from the corresponding time in matched patients, or from the time of treatment failure in patients who remained on a failing regimen. Mortality was analysed using Kaplan-Meier curves and random effects Cox models. RESULTS: Of 16 591 adult patients starting ART, 382 patients (2.3%) switched to a second-line regimen. Another 323 patients (1.9%) did not switch despite developing immunological or virological failure. Cumulative mortality at 1 year was 4.2% (95% CI 2.2-7.8%) in patients who switched to a second-line regimen and 11.7% (7.3%-18.5%) in patients who remained on a failing first-line regimen, compared to 2.2% (1.6-3.0%) in patients on a non-failing first-line regimen (P < 0.0001). Differences in mortality were not explained by nadir CD4 cell count, age or differential loss to follow up. CONCLUSIONS: Many patients who meet criteria for treatment failure do not switch to a second-line regimen and die. There is an urgent need to clarify the reasons why in sub Saharan Africa many patients remain on failing first-line ART. PMID- 20003035 TI - Output that counts: pedometers, sociability and the contested terrain of older adult fitness walking. AB - Based on five months of participant observation and interviews with members of a hospital-sponsored walking club, I explore the rejection of pedometer technology among older adult walkers. Health researchers praise pedometers as a useful tool for measuring walking activity, setting fitness goals, and charting progress towards goals. Older adult walkers, however, viewed pedometers and the monitoring they enable as anathema to walking group norms that stress sociability. I assess the differential construction of pedometers by fitness researchers, group leaders, and walkers themselves. While fitness researchers construct pedometer technology as a motivator for exercise adherence, walkers believed pedometers would create competition and hierarchy that might destroy group camaraderie. In contrast to biomedical models of health and wellness, which focus predominantly on exercise outputs like step counts, these findings suggest that sociability is an important component of health maintenance leisure activities for older adult walkers. PMID- 20003037 TI - In the vanguard of biomedicine? The curious and contradictory case of anti-ageing medicine. AB - The rise of anti-ageing medicine is emblematic of the current conditions of American biomedicine. Through in-depth interviews with 31 anti-ageing practitioners, we examine how practitioners strive for-and justify-a model of care that runs counter to what they see as the 'assembly line' insurance-managed industry of healthcare. Their motivation, however, is not merely a reaction to conventional medicine. It is derived from what they see as a set of core beliefs about the role of the physician, the nature of the physician-patient relationship, and the function of biomedicine. We analyse this ideology to underscore how anti-ageing medicine is built on a 'technology of the self', a self in need of constant surveillance, intervention, and maintenance. The ultimate goal is to create an optimal self, not just a self free of illness. A fundamental irony is that, despite their self-presentation and the perception of the public, anti-ageing providers do not use practices that are especially 'high tech' or unconventional. Instead, the management of ageing bodies rests on providers' perceived knowledge of their patients, tailored treatments, and a collaborative pact between the provider and patient. PMID- 20003038 TI - The clinical assessment of patients admitted to hospital following an episode of self-harm: a qualitative study. AB - Based upon qualitative interviews with clinicians responsible for assessing patients admitted to hospital following an episode of self-harm by overdose, this paper documents interactional factors that shape both how clinicians conduct these assessments and their understanding of what 'causes' someone to harm themselves. The analysis first documents how the patients' overdoses are framed as essentially impulsive acts committed in response to adverse circumstances. This is achieved by clinicians' avoidance of any engagement with, or assessment of, a patient's reasons for taking an overdose. Secondly, the analysis considers why the patients' reasons for taking an overdose do not feature in the clinicians' assessments, and shows, by an examination of two deviant cases, that clinicians avoid engaging with patients' motivations so as to: minimise the risk of generating bedside disagreements; avoid the possibility of being drawn into the emotional life of a patient, and shield themselves from the social and economic inequalities that are a feature of many patients' lives. The paper concludes with a discussion of how bedside assessments that do not engage with patients' reasons for taking an overdose might be experienced by the patients themselves and the deleterious effect this could have upon their clinical management. PMID- 20003039 TI - Pre-emptive resistance: patients' participation in diagnostic sense-making activities. AB - In medical clinic visits, patients do more than convey information about their symptoms and problems so doctors can diagnose and treat them. Patients may also show how they have made sense of their health problems and may press doctors to interpret their problems in certain ways. Using conversation analysis, we analyse a practice patients use early in the medical visit to show that relatively benign or commonplace interpretations of their symptoms are implausible. In this practice, which we term pre-emptive resistance, patients raise candidate explanations for their symptoms and then report circumstances that undermine these explanations. By raising candidate explanations on their own and providing evidence against them, patients call for doctors to restrict the range of diagnostic hypotheses they might otherwise consider. However, the practice does not compel doctors to transparently indicate whether they will do so. Patients also display their ability to recognise and weigh the evidence for common, easily remedied causes of their symptoms. By presenting evidence against them, they show doctors the relevance of more serious diagnostic interpretations without pressing for them outright. PMID- 20003036 TI - The experience of infertility: a review of recent literature. AB - About 10 years ago Greil published a review and critique of the literature on the socio-psychological impact of infertility. He found at the time that most scholars treated infertility as a medical condition with psychological consequences rather than as a socially constructed reality. This article examines research published since the last review. More studies now place infertility within larger social contexts and social scientific frameworks although clinical emphases persist. Methodological problems remain but important improvements are also evident. We identify two vigorous research traditions in the social scientific study of infertility. One tradition uses primarily quantitative techniques to study clinic patients in order to improve service delivery and to assess the need for psychological counselling. The other tradition uses primarily qualitative research to capture the experiences of infertile people in a sociocultural context. We conclude that more attention is now being paid to the ways in which the experience of infertility is shaped by social context. We call for continued progress in the development of a distinctly sociological approach to infertility and for the continued integration of the two research traditions identified here. PMID- 20003040 TI - Childbirth embodiment: problematic aspects of current understandings. AB - The experience of childbirth is one of the most corporeal of the human condition. Against a backdrop of profound change in the milieu of birthing over the past 30 years, especially in the developed world, a number of discourses now compete for the status of the safest, most fulfilling birth experience. Supporters of biomedical and 'natural' approaches make their respective claims to those, with obstetricians broadly aligning their professional interests with the former and midwives with the latter. There is mounting evidence that childbearing women's experiences of birth are often shaped in the uneasy space between the two. Within sociological discourse in health, embodiment is a dominant theme but, to date, research has concentrated mainly on new reproductive technologies, and there is a dearth of recent research and theorising around the act of parturition itself. This paper argues that because of this, there has been a polarising tendency in current discourses which is having a largely negative impact on women, professionals and the maternity services. A call is made for an integration of traditional childbirth embodiment theories, mediated through compassionate, relationally focused maternity care, especially when labour complications develop. PMID- 20003041 TI - Ageing in place and technologies of place: the lived experience of people with dementia in changing social, physical and technological environments. AB - Abstract This paper explores the varied meanings and lived experiences of older people with dementia, in relation to everyday technologies in public spaces outside the home, through re-analysis of qualitative data generated from research that focused on the use of technology in supporting people with dementia to carry on with their everyday outside activities. The data have been re-analysed to critically examine how people with memory loss manage being outside in both familiar and unfamiliar public environments. In doing so the authors explore how technologies mediate between the physical and social environment in which people with dementia live. This paper highlights the importance of a neglected space within dementia research, namely the outside public environment. Although the outside environment and activities that take place in that space, for some, are curtailed, for others the physical and social security of familiar environments enables them to carry on with everyday activities in this public realm. Outside space can be both therapeutic and frightening and this paper demonstrates that people with dementia can sometimes feel out of place in public space. It however also shows the variety of ways people with dementia use everyday technologies to manage 'feeling out of place'. PMID- 20003042 TI - Sirolimus-associated acute interstitial nephritis in a renal allograft. PMID- 20003043 TI - Effects of the novel protein kinase C inhibitor AEB071 (Sotrastaurin) on rat cardiac allograft survival using single agent treatment or combination therapy with cyclosporine, everolimus or FTY720. AB - NVP-AEB071 (AEB, sotrastaurin), an oral inhibitor of protein kinase C (PKC), effectively blocks T-cell activation. The immunosuppressive effects of oral AEB were demonstrated in a rat local graft versus host (GvH) reaction and rat cardiac transplantation models. T-cell activation was suppressed by 95% in blood from AEB treated rats, with a positive correlation between T-cell inhibition and AEB blood concentration. In GvH studies, AEB inhibited lymph node swelling dose-dependently (3-30 mg/kg). BN and DA cardiac allografts were acutely rejected within 6-10 days post-transplantation in untreated LEW rats. AEB at 10 and 30 mg/kg b.i.d. prolonged BN graft survival to a mean survival time of 15 and >28 days, and DA grafts to 6.5 and 17.5 days, respectively. In the DA to LEW model, combining a nonefficacious dose of AEB (10 mg/kg b.i.d.) with a nonefficacious dose of cyclosporine, everolimus or FTY720 led to prolonged median survival times (26 days, >68 days and >68 days, respectively). Pharmacokinetic monitoring excluded drug-drug interactions, suggesting synergy. In conclusion, these studies are the first to demonstrate that AEB prolongs rat heart allograft survival safely as monotherapy and in combination with nonefficacious doses of cyclosporine, everolimus or FTY720. Thus, AEB may have the potential to offer an alternative to calcineurin inhibitor-based therapies. PMID- 20003044 TI - Expansion of hepatic progenitor cell in fatty liver graft after living donor liver transplantation. AB - Although it is known that steatotic livers have a reduced ability to regenerate, most individuals with steatosis show generally benign prognosis. We hypothesized that a proliferative blockade in steatotic hepatocytes results in the compensatory expansion of hepatic progenitor cells (HPC) during fatty liver regeneration. Fifty-four cases of living donor liver transplantation (LDLT) with a liver biopsy performed at the postoperative 10th day were examined. HPC were counted by immunofluorescence histochemical dual-staining technique using cytokeratin 7 and Ki-67, and the replicative arrest of hepatocytes was assessed by p21 immunohistochemistry. The degree of ductular proliferation during regeneration 10 days after LDLT correlated both with the degree of steatosis and the number of HPC (P < 0.001). There was no difference in the average number of HPC and the replicative arrest index between donors with or without steatosis before LDLT (P = 0.111 and P = 0.062). However, degree of steatosis correlated with both the expansion of HPC and the replicative arrest index during liver regeneration 10 days after LDLT (P < 0.001 and P < 0.001, respectively). Moreover, increased replicative arrest was strongly associated with HPC expansion (P < 0.001). In conclusion, the compensatory expansion of HPC as a result of impaired hepatocyte replication occurred during steatotic liver regeneration after LDLT. PMID- 20003045 TI - Outcomes of transplants from patients with small renal tumours, live unrelated donors and dialysis wait-listed patients. AB - We report the outcomes of renal transplant patients (n = 43) who received grafts from donors (n = 41) with small (<3 cm) renal tumours removed before transplantation covering the period from May 1996 to September 2007. Patient and graft survival were compared with the outcomes of conventional live unrelated transplants (LURTs) (n = 120) and to patient survival on the transplant waiting list for those who did not receive a kidney during this period (n = 153). Patient survival at 1, 3 and 5 years were 92%, 88% and 88% for recipients of tumourectomized kidneys (TKs), 99%, 97% and 97% for LURTs, and 98%, 92% and 74% for dialysis patients waiting for a deceased donor kidney (log rank score 10.4, P = 0.005). One patient experienced a local tumour recurrence at 9 years following transplantation. This patient declined intervention and is currently under active surveillance. Transplantation of tumourectomized kidneys from patients with small, localized, incidentally detected renal tumours results in similar outcomes to conventional LURTs and confers a significant survival advantage for patients who would otherwise be unable to receive a transplant. PMID- 20003046 TI - Cryoprecipitate prepared from plasma frozen within 24 hours after phlebotomy contains acceptable levels of fibrinogen and VIIIC. AB - BACKGROUND: As blood centers prepare fewer plasma products from female donors to avoid HLA antibodies, the standard starting material for cryoprecipitate (cryo), fresh-frozen plasma (FFP), might become increasingly scarce. A unit of cryo must contain at a minimum 80 IU of VIIIC and 150 mg of fibrinogen. Plasma frozen within 24 hours after phlebotomy (FP24) contains nearly equivalent levels of clotting factors as FFP at the time of thawing and, although not AABB/FDA approved for this purpose, might be a suitable starting material for cryo (cryo24) manufacture. STUDY DESIGN AND METHODS: FP24 was collected from 20 male donors and cryo24 was subsequently prepared according to standard blood center protocols. The cryo24 was then thawed and VIIIC, von Willebrand factor (VWF) antigen, and fibrinogen levels were measured in these cryo24 units and compared to the corresponding levels in 20 randomly selected units of standard cryo (prepared from FFP). RESULTS: The mean (+/-SD) levels in cryo and cryo24 at the time of thawing were as follows: fibrinogen (mg/unit) 455.8 (+/-172.6) and 575.8 (+/-185.9; p = 0.04), VIIIC (IU/unit) 216.1 (+/-52.3) and 252.4 (+/-70.1; p = 0.07), and VWF (IU/unit) 448.1 (+/-118.9) and 505.9 (+/-135.1; p = 0.16), respectively. All units of standard cryo and cryo24 met the current minimum content standards. CONCLUSION: All units of cryo24 in this pilot study met the current minimum content standards for cryo. While clinical studies of its efficacy are warranted, cryo24's in vitro characteristics are at least equivalent to that of standard cryo. PMID- 20003047 TI - Prolonged anemia in an intrauterine-transfused neonate with Rh-hemolytic disease: no evidence for anti-D-related suppression of erythropoiesis in vitro. AB - BACKGROUND: Rh-hemolytic disease may be complicated in some cases by a prolonged postnatal anemia with an extended need for postnatal red blood cell (RBC) transfusion. Besides ongoing hemolysis, marrow suppression and erythropoietin (EPO) deficiency are discussed as underlying mechanisms of this so-called "late hyporegenerative anemia." CASE REPORT: We present a case of a newborn with Rh hemolytic disease caused by anti-D who received several intrauterine RBC transfusions. After birth, no reticulocytes or D+ RBCs were detectable in peripheral blood of the infant; thus further RBC transfusions were necessary. Administration of intravenous immunoglobulins had no obvious effect. Reticulocytes first became detectable 15 weeks after birth, when anti-D titer had decreased to 16. A few days later, hemoglobin started to increase and no further treatment was necessary. To investigate whether anti-D is able to cause maturation arrest of erythroid progenitors, maternal serum was added to an in vitro assay of erythropoiesis, induced from human CD34+ cells. RESULTS: In this case, no variables of hemolysis (e.g., elevated bilirubin) were observed. The EPO level was normal and a marrow sample showed increased erythropoiesis. The in vitro erythropoiesis assay revealed no influence of anti-D on RBC proliferation and differentiation. CONCLUSION: Anemia in our patient seemed to be mainly caused by ongoing intramedullar hemolysis due to persistent high anti-D titers. In such cases, variables for hemolysis are not necessarily found. Release of patient's own RBCs into the circulation may become sufficient when anti-D has declined to a very low level of approximately 16. PMID- 20003048 TI - A dual antiglobulin test for the detection of weak or nonagglutinating immunoglobulin M warm autoantibodies. AB - BACKGROUND: Immunoglobulin (Ig)M warm autoantibodies (AABs) usually cause severe autoimmune hemolytic anemia (AIHA) and, in some cases, red blood cell (RBC)-bound IgM cannot be detected. We describe a simple dual antiglobulin test (DDAT) for diagnosing such cases. STUDY DESIGN AND METHODS: A patient with erroneously suspected cold agglutinin syndrome was investigated. The direct antiglobulin test (DAT) was performed using standard techniques and dual (two stages) antiglobulin reagents (IgG rabbit anti-human IgM, IgG goat anti-rabbit IgG). RESULTS: A cold agglutinin syndrome was diagnosed initially, as the patient's serum was reactive with RBCs at a temperature of 28 degrees C or less, and the DAT was strongly positive with anti-C(3)d. Six months later, the patient was reexamined at this hospital due to progressive hemolysis. His RBCs were found to be coated with IgM warm AABs that only became detectable using a DDAT, and his serum contained only a weak cold agglutinin. The hemolysis remained refractory to treatment with prednisolone and also prednisolone plus azathioprine, but gradually improved after treatment with prednisolone plus cyclophosphamide. CONCLUSION: Weak or nonagglutinating RBC-bound IgM warm antibodies can be identified by the presented DDAT. PMID- 20003049 TI - Prolonged elevation of plasma argatroban in a cardiac transplant patient with a suspected history of heparin-induced thrombocytopenia with thrombosis. AB - BACKGROUND: Direct thrombin inhibitors (DTIs) provide an alternative method of anticoagulation for patients with a history of heparin-induced thrombocytopenia (HIT) or HIT with thrombosis (HITT) undergoing cardiopulmonary bypass (CPB). In the following report, a 65-year-old critically ill patient with a suspected history of HITT was administered argatroban for anticoagulation on bypass during heart transplantation. The patient required massive transfusion support (55 units of red blood cells, 42 units of fresh-frozen plasma, 40 units of cryoprecipitate, 40 units of platelets, and three doses of recombinant Factor VIIa) for severe intraoperative and postoperative bleeding. STUDY DESIGN AND METHODS: Plasma samples from before and after CPB were analyzed postoperatively for argatroban concentration using a modified ecarin clotting time (ECT) assay. RESULTS: Unexpectedly high concentrations of argatroban were measured in these samples (range, 0-32 microg/mL), and a prolonged plasma argatroban half life (t(1/2)) of 514 minutes was observed (published elimination t(1/2) is 39-51 minutes [< or = 181 minutes with hepatic impairment]). CONCLUSIONS: Correlation of plasma argatroban concentration versus the patient's coagulation variables and clinical course suggest that prolonged elevated levels of plasma argatroban may have contributed to the patient's extended coagulopathy. Because DTIs do not have reversal agents, surgical teams and transfusion services should remain aware of the possibility of massive transfusion events during anticoagulation with these agents. This is the first report to measure plasma argatroban concentration in the context of CPB and extended coagulopathy. PMID- 20003050 TI - Considerations on the use of adjunct red blood cell exchange transfusion in the treatment of severe Plasmodium falciparum malaria. AB - BACKGROUND: Travelers returning to the United States from malaria-endemic areas are at increased risk of a potentially fatal infection from Plasmodium falciparum, which requires prompt and aggressive treatment. STUDY DESIGN AND METHODS: Described is a case of a 7-year-old boy who was infected by P. falciparum while in Africa and developed features of severe infection, including hyperparasitemia, altered neurologic status, and malarial hepatitis. RESULTS: A single automated erythrocytapheresis procedure reduced parasitemia from 14% to less than 1%. Along with intravenous quinidine, this reduced parasite level was maintained throughout the hospitalization and the patient recovered. CONCLUSION: Exchange transfusion (ET) is an effective adjunct therapy to reduce the parasite load in cases of severe P. falciparum malaria. When performed in certain defined settings, the benefits can outweigh the risks of the procedure. Discussed are the medical and technical considerations on the use of adjunctive ET for severe P. falciparum infection and a review of the literature of the use of adjunct ET in the treatment of severe P. falciparum malaria. PMID- 20003051 TI - Demographic profile of blood donors at three major Brazilian blood centers: results from the International REDS-II study, 2007 to 2008. AB - BACKGROUND: The profile of blood donors changed dramatically in Brazil over the past 20 years, from remunerated to nonremunerated and then from replacement to community donors. Donor demographic data from three major blood centers establish current donation profiles in Brazil, serving as baseline for future analyses and tracking longitudinal changes in donor characteristics. STUDY DESIGN AND METHODS: Data were extracted from the blood center, compiled in a data warehouse, and analyzed. Population data were obtained from the Brazilian census. RESULTS: During 2007 to 2008, there were 615,379 blood donations from 410,423 donors. A total of 426,142 (69.2%) were from repeat (Rpt) donors and 189,237 (30.8%) were from first-time (FT) donors. Twenty percent of FT donors returned to donate in the period. FT donors were more likely to be younger, and Rpt donors were more likely to be community donors. All were predominantly male. Replacement donors still represent 50% of FT and 30% of Rpt donors. The mean percentage of the potentially general population who were donors was approximately 1.2% for the three centers (0.7, 1.5, and 3.1%). Adjusting for the catchment's area, the first two were 2.1 and 1.6%. CONCLUSIONS: Donors in the three Brazilian centers tended to be younger with a higher proportion of males than in the general population. Donation rates were lower than desirable. There were substantial differences in sex, age, and community/replacement status by center. Studies on the safety, donation frequencies, and motivations of donors are in progress to orient efforts to enhance the availability of blood. PMID- 20003052 TI - Thrombotic thrombocytopenic purpura at the Johns Hopkins Hospital from 1992 to 2008: clinical outcomes and risk factors for relapse. AB - BACKGROUND: Plasma exchange, the standard treatment for thrombotic thrombocytopenic purpura (TTP), has significantly decreased disease mortality. However, TTP recurs in 20% to 50% of patients who survive the initial episode. We aimed to describe the clinical spectrum of TTP, to determine the valid endpoint for plasma exchange cessation, and to explore the risk factors for disease relapse. STUDY DESIGN AND METHODS: Using the ICD-9 diagnosis code, we identified patients treated for TTP at the Johns Hopkins Hospital between 1992 and 2008. Complete demographic, clinical, laboratory, treatment, and outcome data were collected from the medical records. RESULTS: A total of 72 patients were treated for 134 episodes of TTP at the Johns Hopkins Hospital during the study period. With standardized combined immunosuppression and plasma exchange treatment, the all-cause mortality rate was 4%. Lactate dehydrogenase (LDH) normalization lagged behind platelet (PLT) recovery by an average of 9 days and did not predict response. Relapse occurred in 36% of patients during a median follow-up of 30 months with most (76%) occurring in the first 24 months. African American ethnicity was associated with increased risk of relapse (odds ratio = 4.8, p = 0.03). CONCLUSIONS: Excellent outcomes in patients with TTP are achievable with multimodality therapy. LDH normalization lags behind PLT recovery and might not be an informative endpoint for plasma exchange cessation. Prospective studies are warranted to confirm the influence of race on relapse and identify additional risk factors for adverse outcomes that could be targeted to improve therapeutic outcomes for patients with TTP. PMID- 20003053 TI - Impact of using different laboratory assays to detect human leukocyte antigen antibodies in female blood donors. AB - BACKGROUND: HLA antibodies passively transferred to transfused recipients may cause transfusion reactions such as transfusion-related acute lung injury (TRALI), but in many of the reported TRALI incidents, no white blood cell antibodies have been identified. We investigated whether a higher number of anti HLA would be detected in donor's plasma by using a method with potential higher sensitivity rate. STUDY DESIGN AND METHODS: Sera from 300 previously pregnant female blood donors were screened for anti-HLA using a solid-phase mixed-antigen assay (enzyme-linked immunosorbent assay [ELISA]). Samples from 60 women with three or more pregnancies with a negative ELISA were further tested using microbead-flow assays (LABScreen mixed, panel-reactive antibodies [PRA], and single antigen). RESULTS: Anti-HLA Class I and/or Class II were detected by ELISA in 26.7% (80/300) of all women and in 37.0% (37/100) of women with three or more pregnancies. The LABScreen assays detected additional anti-HLA specificities (44 Class I and 17 Class II) in 28.3% (17/60) of ELISA-negative donors with three or more pregnancies. HLA antibodies were detected in 8.3% (5/60), 18.3% (11/60), and 21.7% (13/60) of ELISA-negative women by LABScreen mixed, PRA, or single antigen, respectively. CONCLUSION: Our data showed that the microbead-flow detected more HLA antibodies than ELISA, but the clinical significance of these antibodies is currently unknown. Detecting anti-HLA is useful for donor management and could contribute to the decision to definitively defer blood donors involved in TRALI incidents. However, further studies are necessary to better determinate the relative risk of TRALI induced by anti-HLA detected only by techniques with higher sensitivity rate. PMID- 20003054 TI - Evaluating maturation and genetic modification of human dendritic cells in a new polyolefin cell culture bag system. AB - BACKGROUND: Dendritic cells (DCs) are applied worldwide in several clinical studies of immune therapy of malignancies, autoimmune diseases, and transplantations. Most legislative bodies are demanding high standards for cultivation and transduction of cells. Closed-cell cultivating systems like cell culture bags would simplify and greatly improve the ability to reach these cultivation standards. We investigated if a new polyolefin cell culture bag enables maturation and adenoviral modification of human DCs in a closed system and compare the results with standard polystyrene flasks. STUDY DESIGN AND METHODS: Mononuclear cells were isolated from HLA-A*0201-positive blood donors by leukapheresis. A commercially available separation system (CliniMACS, Miltenyi Biotec) was used to isolate monocytes by positive selection using CD14-specific immunomagnetic beads. The essentially homogenous starting cell population was cultivated in the presence of granulocyte-macrophage-colony-stimulating factor and interleukin-4 in a closed-bag system in parallel to the standard flask cultivation system. Genetic modification was performed on Day 4. After induction of maturation on Day 5, mature DCs could be harvested and cryopreserved on Day 7. During the cultivation period comparative quality control was performed using flow cytometry, gene expression profiling, and functional assays. RESULTS: Both flasks and bags generated mature genetically modified DCs in similar yields. Surface membrane markers, expression profiles, and functional testing results were comparable. The use of a closed-bag system facilitated clinical applicability of genetically modified DCs. CONCLUSIONS: The polyolefin bag-based culture system yields DCs qualitatively and quantitatively comparable to the standard flask preparation. All steps including cryopreservation can be performed in a closed system facilitating standardized, safe, and reproducible preparation of therapeutic cells. PMID- 20003055 TI - Efficient generation of clinical-grade genetically modified dendritic cells for presentation of multiple tumor-associated proteins. AB - BACKGROUND: Dendritic cells (DCs) play a central role in the initiation and regulation of immune responses. DCs for clinical applications can be generated with high yield from leukapheresis products. Using adenoviral transduction we genetically modified human DCs to produce and present melanoma-associated antigens. Coexpression of green fluorescent protein and epitope tags were used to monitor genetic modification. Generation, genetic modification, and cryoconservation of gene modified human DCs on a clinical scale in a closed system is feasible. STUDY DESIGN AND METHODS: CD14-positive monomuclear cells were isolated from leukapheresis products of HLA-A* 0201 positive voluntary blood donors using immunomagnetic beads. Selected cells were cultivated for 7 days. Adenovirus transduction was optimal on Day 4. Maturation was induced on Day 5. Mature DC were aliquoted and cryoconserved on Day 7. Quality control was performed using flow cytometry, expression profiling, and functional assays (ELISPOT, CBA). RESULTS: We were able to generate sufficient genetically modified mature DCs in serum-free cultures that could be stored by cryopreservation. The use of a closed system facilitated development of methods for standardized production of clinically applicable genetically modified DCs. The adenoviral transduction system allowed simultaneous and flexible expression of tumor associated antigens for prolonged presentation of multiple epitopes. CONCLUSION: The feasibility of a closed-bag system for the cultivation of genetically modified human DCs is shown. The immature DCs were genetically modified by recombinant replication-deficient adenoviruses to express multiple epitopes of tumor-associated proteins and then differentiated to mature antigen-presenting DCs. PMID- 20003056 TI - The impact of simple donor education on donor behavioral deferral and infectious disease rates in Sao Paulo, Brazil. AB - BACKGROUND: Studies have shown that human immunodeficiency virus (HIV) residual risk is higher in Brazilian than in US and European blood donors, probably due to failure to defer at-risk individuals in Brazil. This study assessed the impact of an educational brochure in enhancing blood donors' knowledge about screening test window phase and reducing at-risk individuals from donating. STUDY DESIGN AND METHODS: This trial compared an educational intervention with a blood center's usual practice. The brochure was distributed in alternating months to all donors. After donating, sampled participants completed two questions about their HIV window period knowledge. The impact on HIV risk deferral, leaving without donation, confidential unit exclusion (CUE) use, and test positivity was also analyzed. RESULTS: From August to November 2007 we evaluated 33,940 donations in the main collection center of Fundacao Pro-Sangue/Hemocentro de Sao Paulo in Sao Paulo, Brazil. A significant (p < 0.001) pamphlet effect was found on correct responses to both questions assessing HIV window phase knowledge (68.1% vs. 52.9%) and transfusion risk (91.1% vs. 87.2%). After adjusting for sex and age, the pamphlet effect was strongest for people with more than 8 years of education. There was no significant pamphlet effect on HIV risk deferral rate, leaving without donation, use of CUE, or infectious disease rates. CONCLUSION: While the educational pamphlet increased window period knowledge, contrary to expectations this information alone was not enough to make donors self-defer or acknowledge their behavioral risk. PMID- 20003057 TI - Evaluation of removal of prion infectivity from red blood cells with prion reduction filters using a new rapid and highly sensitive cell culture-based infectivity assay. AB - BACKGROUND: The clearance of infectious prions from biologic fluids is usually quantified by bioassays based on intracerebral inoculation of hamsters or mice; these tests are slow, cumbersome, imprecise, and very expensive. In the present study we describe the use of a new and highly sensitive cell culture-based infectivity assay to evaluate the performance of several prion removal prototype filters. STUDY DESIGN AND METHODS: Five units of 1- to 2-day-old ABO-compatible human red blood cells (RBCs) in saline-adenine-glucose-mannitol were obtained from an AABB-accredited blood bank. The 5 units were combined to create a homogenous pool. Scrapie-infected mouse brain homogenate of a Rocky Mountain Laboratory strain was added to the pooled RBCs. The pooled RBCs were divided into 300-mL aliquots, which were filtered with either standard leukoreduction filter or four prototypes of prion reduction filter. The levels of prion infectivity in the pre- and postfiltration samples were measured with a cell culture-based standard scrapie cell assay (SSCA). RESULTS: All the 22-layer prion reduction filters removed prion infectivity below the limit of detection of the SSCA (reduction in prion infectivity > or =2.0 log(10)LD(50)/mL) while the 10-layer variant showed some residual infectivity. CONCLUSIONS: These results demonstrate the utility of a highly sensitive cell culture-based infectivity assay for screening prion reduction filters. The use of this type of in vitro infectivity assay will substantially help expedite the screening and discovery of devices aimed at reducing the risk of variant Creutzfeldt-Jakob disease transmission through blood transfusion. PMID- 20003058 TI - Procedure-specific preoperative red blood cell preparation and utilization management in pediatric surgical patients. AB - BACKGROUND: Data-driven practices in preoperative red blood cell (RBC) preparation for pediatric surgical procedures are not well established. Adaptation of established adult preparation guidance methods to pediatric populations may improve perioperative RBC utilization. STUDY DESIGN AND METHODS: A retrospective audit of preoperative RBC preparation volumes (Vp) and intraoperative RBC transfusion volumes (Vt) for pediatric surgical procedures was undertaken at a large children's hospital from January to June 2006. RBC preparation-to-transfusion volume (mL/kg) ratios (P:T) were calculated for all surgeries, subspecialties, and select procedures. P:T equals Vp divided by Vt. Resulting P:Ts were compared to a target P:T of 2:1. A model for maximum procedure-specific Vp (Vp-max) defined Vp-max as the RBC transfusion volume able to meet the needs of 80% of patients undergoing an individual surgical procedure. Vp-max values were applied to the study data set to predict the impact on P:Ts and Vp. RESULTS: RBCs were prepared for 332 surgical procedures and transfused during 113 procedures. P:T was 3.5:1 for total surgical procedures (subspecialty range, 2.7:1-46:0), exceeding the 2:1 target. Vp-max modeling for spinal fusion, craniotomy for neoplasia, craniotomy for seizure, and craniosynostectomy yielded P:T ratios of 1.5:1, 1.5:1, 1.7:1, and 1.0:1, respectively, predicting a 30% decrease in Vp for these four surgical procedures. CONCLUSIONS: P:Ts for pediatric surgical procedures at this institution indicate potentially excessive preoperative RBC preparations. Determination of data-driven procedure-specific Vp may increase the efficiency of preoperative RBC preparation practices. PMID- 20003059 TI - Novel single-nucleotide change in GYP*A in a person who made an alloantibody to a new high-prevalence MNS antigen called ENEV. AB - BACKGROUND: Alloantibodies that define some high-prevalence MNS antigens are made by people with glycophorin A (GPA) altered by a single-amino-acid change or replacement of amino acids from part of the Pseudoexon 3 of GYP*B. The finding of a patient whose plasma contained a novel alloanti-En(a)FR prompted this study. RESULTS: The patient's serum contained an alloantibody to a high-prevalence antigen, resistant to papain, ficin, trypsin, alpha-chymotrypsin, or dithiothreitol. The antibody was strongly reactive with all panel red blood cells (RBCs) tested, showed reduced reactivity with ENEP- and ENAV- RBCs, and was nonreactive with M(k)M(k), En(a-), GP.Hil/GP.Hil, and GP.JL/M(k) RBCs. The patient's RBCs typed M+N-S+s-, Wr(a-b+(w)), ENEP-, and ENAV-. These results indicated that the antibody recognized a new high-prevalence antigen in the MNS system. Sequencing of DNA prepared from the patient's white blood cells revealed a GYP*A nucleotide substitution of 242T>G (predicted to change Val62 of GPA to Gly). This change ablates an RsaI restriction enzyme site and polymerase chain reaction-restriction fragment length polymorphism confirmed that the proband was homozygous for Nucleotide 242G. CONCLUSIONS: We describe a novel high-prevalence MNS antigen, characterized by Val62 in GPA and named ENEV. The absence of the antigen is associated with Gly62. The change explains the weakened reactivity of the patient's serum with ENEP- and ENAV- RBCs and nonreactivity with anti-ENEP and anti-ENAV against her RBCs. The ENEV antigen has been assigned the ISBT number MNS45. PMID- 20003060 TI - Is red blood cell rheology preserved during routine blood bank storage? AB - BACKGROUND: Red blood cell (RBC) units stored for more than 2 weeks at 4 degrees C are currently considered of impaired quality. This opinion has primarily been based on altered RBC rheologic properties (i.e., enhanced aggregability, reduced deformability, and elevated endothelial cell interaction), during prolonged storage of nonleukoreduced RBC units. In this study, the rheologic properties and cell variables of leukoreduced RBC units, during routine blood bank storage in saline-adenine-glucose-mannitol, were investigated. STUDY DESIGN AND METHODS: Ten leukoreduced RBC units were stored at the blood bank for 7 weeks at 4 degrees C. RBCs were tested weekly for aggregability, deformability, and other relevant variables. RESULTS: RBC aggregability was significantly reduced after the first week of storage but recovered during the following weeks. After 7 weeks aggregability was slightly, but significantly, reduced (46.9 + or - 2.4-44.3 + or - 2.2 aggregation index). During storage the osmotic fragility was not significantly enhanced (0.47 + or - 0.01% phosphate-buffered saline) and the deformability at shear stress of 3.9 Pa was not significantly reduced (0.36 + or 0.01 elongation index [EI]). The deformability at 50 Pa was reduced (0.58 + or - 0.01-0.54 + or - 0.01 EI) but remained within reference values (0.53 + or - 0.04). During 5 weeks of storage, adenosine triphosphate was reduced by 54% whereas mean cell volume, pH, and mean cell hemoglobin concentration were minimally affected. CONCLUSIONS: RBC biochemical and physical alterations during storage minimally affected the RBC ability to aggregate and deform, even after prolonged storage. The rheologic properties of leukoreduced RBC units were well preserved during 7 weeks of routine blood bank storage. PMID- 20003061 TI - Activity-based costs of blood transfusions in surgical patients at four hospitals. AB - BACKGROUND: Blood utilization has long been suspected to consume more health care resources than previously reported. Incomplete accounting for blood costs has the potential to misdirect programmatic decision making by health care systems. Determining the cost of supplying patients with blood transfusions requires an in depth examination of the complex array of activities surrounding the decision to transfuse. STUDY DESIGN AND METHODS: To accurately determine the cost of blood in a surgical population from a health system perspective, an activity-based costing (ABC) model was constructed. Tasks and resource consumption (materials, labor, third-party services, capital) related to blood administration were identified prospectively at two US and two European hospitals. Process frequency (i.e., usage) data were captured retrospectively from each hospital and used to populate the ABC model. RESULTS: All major process steps, staff, and consumables to provide red blood cell (RBC) transfusions to surgical patients, including usage frequencies, and direct and indirect overhead costs contributed to per-RBC-unit costs between $522 and $1183 (mean, $761 +/- $294). These exceed previously reported estimates and were 3.2- to 4.8-fold higher than blood product acquisition costs. Annual expenditures on blood and transfusion-related activities, limited to surgical patients, ranged from $1.62 to $6.03 million per hospital and were largely related to the transfusion rate. CONCLUSION: Applicable to various hospital practices, the ABC model confirms that blood costs have been underestimated and that they are geographically variable and identifies opportunities for cost containment. Studies to determine whether more stringent control of blood utilization improves health care utilization and quality, and further reduces costs, are warranted. PMID- 20003062 TI - Ultra-short course immunotherapy in children and adolescents during a 3-yrs post marketing surveillance study. AB - Ultra-short course immunotherapy (uSCIT) has shown good efficacy and tolerability in children and adolescents with seasonal allergic rhinitis (SAR), conjunctivitis and/or asthma in clinical studies. Here, we investigate the efficacy of uSCIT in the juvenile subpopulation of a 3-year post-marketing surveillance (PMS) study. To assess the differences in the efficacy of uSCIT between adults and children respectively adolescents enrolled in a PMS study. In a prospective open study 422 patients aged 6-18 years with SAR, conjunctivitis and/or asthma received four pre seasonal injections with pollen allergoids formulated with monophosphoryl lipid A (MPL, Pollinex Quattro) over a minimum of 3 weeks. Efficacy was evaluated by response to therapy and consumption of anti-allergic medication during the pollen season. Tolerability was evaluated by patients' acceptance of therapy. These results were compared with the adult subpopulation of this study. Response to treatment was assessed as good or very good in 94% of patients, mirroring findings for the entire cohort. Further improvements were noted in patients receiving subsequent courses of therapy. Anti-allergic medication use decreased from 83% to 24% of patients after the first treatment course (p < 0.0001). Therapy was well accepted by children/adolescents and considered 'very good' or 'good' by 93% of juveniles. No serious adverse events or cases of anaphylaxis were reported. This subanalysis indicated that uSCIT with Pollinex Quattro had similar efficacy and tolerability in children/adolescents and adults. The convenient dosing regimen and favourable safety profile of uSCIT may support a wider uptake of uSCIT in this patient population. PMID- 20003063 TI - Polycyclic aromatic hydrocarbon metabolite levels and pediatric allergy and asthma in an inner-city cohort. AB - Exposure to polycyclic aromatic hydrocarbons (PAH) has been associated with allergic sensitization and asthma. We hypothesized that increased urinary PAH metabolites are associated with allergy or asthma among children age 5 yrs in an inner-city birth cohort. As part of an ongoing prospective birth cohort under the auspices of the Columbia Center for Children's Environmental Health (CCCEH), urine was collected from 5-yr-old children (n = 222) of Dominican American and African American mothers in Northern Manhattan and South Bronx of New York City. Twenty-four PAH metabolites were measured in these specimens, and their levels (unadjusted and specific gravity corrected) were evaluated with IgE levels and asthma outcomes. Ten metabolites were detected in urine from all children. Concentrations ranged higher than those in representative samples of US children ages 6-11 in the National Health and Nutrition Examination Survey (NHANES). Among CCCEH children, compared with African Americans, the Dominican children had higher 2-hydroxynaphthalene but lower 9-hydroxyfluorene and 4-hydroxyphenanthrene concentrations. Increased 3-hydroxyfluorene and 3-hydroxyphenanthrene levels were associated with higher anti-mouse IgE levels (p < 0.05). These plus 2 hydroxynaphthalene, 2-hydroxyflourene and 1-hydroxyphenanthrene concentrations were associated with higher anti-mouse IgE levels on multivariate analyzes. Increased 2-hydroxyphenanthrene, 3-hydroxyphenanthrene and 4-hydroxyphenanthrene levels were associated with higher anti-cat IgE levels (p < 0.05) in univariate, but not multivariate, analyzes. Levels of PAH metabolites were not associated with respiratory symptoms. Measures of PAH metabolites suggest considerable exposure in an urban pediatric population, and possible associations with allergic sensitization to mouse. PMID- 20003064 TI - Exploration of basal immune parameters in healthy infants receiving an infant milk formula supplemented with prebiotics. AB - This double-blind, randomized, placebo-controlled study, aimed to explore the effect of an infant milk formula (IMF) with 6 g/l short-chain galacto- and long chain fructo-oligosaccharides (scGOS/lcFOS, ratio 9:1) on basal immune parameters in 215 healthy, term infants during the first 26 wk of life. After birth, the infants received breast milk or were randomized to receive an IMF with or without scGOS/lcFOS. Blood samples were collected at the age of 8 wk and 26 wk for the analysis of serum immunoglobulins, lymphocyte subpopulations, and cytokines. The scGOS/lcFOS group and the control group were compared in the statistical analysis. A breast fed group was included as a reference. In total, 187 Infants completed the study. No significant differences were observed between both formula groups in the different studied immune parameters at weeks 8 and 26. This explorative study indicates that supplementation of infant formula with a mixture of prebiotic oligosaccharides did not change the basal level of the measured parameters of the developing immune system in healthy infants with a balanced immune system during the first 6 months of life in comparison to feeding a standard infant formula and in comparison to exclusive breastfeeding. PMID- 20003065 TI - Determination of multiple allergen-specific IgE by microfluidic immunoassay cartridge in clinical settings. AB - Our aims were to evaluate the performance of an automated microfluidic immunoassay system for measuring allergen-specific IgE (sIgE) in sera against an established in vitro assay and to assess the system's diagnostic accuracy against objective clinical criteria for identifying sensitization to specific allergens in daily practice of allergy clinics. Using both the automated microfluidic-based immunoassay system (BioIC and ImmunoCAP, we measured sIgE in serum samples from 212 children who visited allergic clinics in two medical centers. Outcomes of skin prick tests (SPT) served as the clinical comparison method. The assay results of targeted allergen of BioIC have a good correlation with ImmunoCAP in the diagnosis of allergen sensitivity by patients' clinical history. When comparing the test results of the sIgE against overall allergens, in either two tests among the three assays performed showed high percentage of agreement between BioIC and ImmunoCAP (77.8%, 95% CI: 72-83.3%) but not with SPT (BioIC 64.9%, 95% CI: 58-72%; ImmunoCAP 67.5%, 95% CI: 61-74%). Using ROC analysis and SPT as quasi-standard, BioIC and ImmunoCAP have nearly the same performance of sensitivity and specificity in the confirmation of SPT results. The total and within one-class agreements of each allergen test result between BioIC and ImmunoCAP ranged between 55.2% and 99.5% with an overall average of 80.9%. Laboratory testing for sIgE can be performed on a fully automated, microfluidic cartridge system with advantages of low sample volume, simultaneously tested allergens, and with diagnostic accuracy for representative allergens equivalent to the semi-automated CAP technology. PMID- 20003066 TI - Probiotics for allergic diseases: realities and myths. AB - The prevalence of allergic diseases such as asthma, allergic rhinitis, and atopic dermatitis has increased sharply over the past two to three decades in many countries, and allergies are now the most common chronic disease among children throughout the world. In the past few years, probiotics have been advocated for the management of allergic diseases in many parts of the world. Physicians have a responsibility to ensure the efficacy and safety of any products they prescribe or recommend. This article provides a comprehensive overview and a critical interpretation of currently available evidence regarding the role of probiotics in the prevention and treatment of allergic diseases in humans and also discusses several major myths and potential risks associated with the use of probiotics. In the current era of evidence-based medicine, there is still insufficient evidence to recommend probiotics for the prevention of allergic diseases or as part of standard management for any allergic conditions in children. PMID- 20003067 TI - Allergy related disorders among 2-yrs olds in a general population. The PACT Study. AB - Allergic disorders represent a major health problem in most developed countries, but few population-based studies have focused on these disorders in early childhood. The aims of the present study were to investigate the prevalence, gender differences and distribution of allergy related disorders and their association to sensitization among unselected children, 2 yrs of age, in a general population. A population-based study with parental self reported questionnaire data involving allergy related symptoms and results from allergy tests from 4783 two-yr-old children was conducted, and skin prick tests (SPT) of a randomly selected sample comprising 390 children were performed. In the total population the prevalence of reported wheeze was 26%, doctor diagnosed asthma (DDAsthma) 7.0%, atopic dermatitis (AD) 17% and allergic rhinoconjunctivitis (ARC) 3%. Of the 1008 (21%) allergy tested children 59% reported a positive test, but of the randomly selected children only 8% had a positive SPT. Children with AD were most frequently sensitized and children with ARC were most likely to have other allergy related disorders (70%). More boys than girls had an allergy related disorder or a positive allergy test. In conclusion, two in five had an allergy related disorder, but less than 10% had a positive SPT. Having one allergic disorder, especially ARC, increased substantially the risk of having another, and having AD was most strongly associated to a positive allergy test. Moreover, boys were more likely than girls to have an allergy related disorder or a positive SPT indicating a gender difference in the natural history of allergy related disorders. PMID- 20003068 TI - Maternal diet during pregnancy and allergic sensitization in the offspring by 5 yrs of age: a prospective cohort study. AB - To examine the effect of maternal diet during pregnancy on allergic sensitization in the offspring by 5 yrs of age. The Finnish type 1 Diabetes Prediction and Prevention Nutrition Study. A population-based cohort study with 5-yr follow-up. A total of 931 children with human leukocyte antigen-conferred susceptibility to type 1 diabetes for whom maternal pregnancy food frequency questionnaire data and allergen-specific immunoglobulin E measurement at 5 yrs were available. Increasing maternal consumption of citrus fruits [odds ratio (OR) = 1.14, 95% confidence interval (CI) = 1.05-1.25] and total fruit (OR = 1.36, 95% CI = 1.09 1.70) were positively associated with sensitization to inhalant allergens, after adjustment for potential confounders. Maternal intake of vitamin D (OR = 0.56, 95% CI = 0.35-0.91) was inversely associated with sensitization to food allergens. Maternal consumption of citrus fruits during pregnancy may increase the risk to allergic sensitization in the offspring, whereas vitamin D intake may have a beneficial effect. Further studies are required to define more closely the putative effect of maternal intake of polyunsaturated fatty acids on development of allergic diseases in the offspring. PMID- 20003069 TI - Treatment of clinical insulin resistance in children: a systematic review. AB - The objective of this study was to evaluate the effectiveness of interventions aimed at improving clinical insulin resistance and/or pre-diabetes in children. This study is a systematic review and meta-analysis. Five electronic databases were searched for randomized controlled trials of at least 2-months' duration. The outcomes were fasting insulin, homeostasis model assessment of insulin resistance (HOMA-IR), body mass index (BMI) and adverse outcomes. Four randomized controlled trials were identified. All compared the effect of 6 months of metformin plus or minus lifestyle intervention with placebo plus or minus lifestyle intervention. After pooling results from three trials, the mean difference after 6 months favoured the intervention with a statistically significant mean decrease in fasting insulin, HOMA-IR and BMI of 9.6 uU mL(-1) (95% confidence interval [CI]: 6.3, 13.0 uU mL(-1) ; I(2) = 76%), 2.7 (95% CI: 1.7, 3.6; I(2) = 74%) and 1.7 kg m(-2) (95% CI: 1.1, 2.3 kg m(-2) ; I(2) = 75) respectively. Mild gastrointestinal symptoms were reported in 19% (2-29%; median and range) of participants taking metformin. Metformin improves markers of insulin sensitivity and reduces BMI in children and adolescents with clinical insulin resistance or pre-diabetes. Stronger evidence from high-quality studies of longer duration and larger sample size are required before clinical conclusions about the optimal treatment protocol in this population can be drawn. PMID- 20003070 TI - Percentile distributions of waist circumference for 7-19-year-old Polish children and adolescents. AB - The aim of the study was to develop waist circumference (WC) percentiles in Polish children and youth and to compare these with the results obtained in other countries. The study comprised a random group of 5663 Polish children aged 7-18 years. Smoothed WC percentile curves were computed using the LMS method. The curves displaying the values of the 50th (WC(50)) and the 90th (WC(90)) percentile were then compared with the results of similar studies carried out in children from the UK, Spain, Germany, Turkey, Cyprus, Canada and the USA. WC increased with age in both boys and girls and in all observed age periods the boys were seen to dominate. For 18-year-old Polish boys and girls the values of WC(90) were 86.5 and 78.2, respectively, and were lower than the current criteria developed by the International Diabetes Federation. Both WC(50) and WC(90) were higher in Polish boys and girls compared with their counterparts in the UK, Turkey and Canada and significantly lower than in children from the USA, Cyprus and Spain. The percentile curves for Polish children and youth, which were developed here for the first time, are base curves that can be applied in analysing trends as well as making comparisons with results of similar studies performed in other countries. PMID- 20003071 TI - Resistance training for obese, type 2 diabetic adults: a review of the evidence. AB - In both developed and developing countries, increased prevalence of obesity has been strongly associated with increased incidence of type 2 diabetes mellitus (T2DM) in the adult population. Previous research has emphasized the importance of physical activity in the prevention and management of obesity and T2DM, and generic exercise guidelines originally developed for the wider population have been adapted for these specific populations. However, the guidelines traditionally focus on aerobic training without due consideration to other exercise modalities. Recent reviews on resistance training in the T2DM population have not compared this modality with others including aerobic training, or considered the implications of resistance training for individuals suffering from both obesity and T2DM. In short, the optimal mix of exercise modalities in the prescription of exercise has not been identified for it benefits to the metabolic, body composition and muscular health markers common in obesity and T2DM. Similarly, the underlying physical, social and psychological barriers to adopting and maintaining exercise, with the potential to undermine the efficacy of exercise interventions, have not been addressed in earlier reviews. Because it is well established that aerobic exercise has profound effects on obesity and T2DM risk, the purpose of this review was to address the importance of resistance training to obese adults with T2DM. PMID- 20003072 TI - Stress-induced DNA methylation changes and their heritability in asexual dandelions. AB - *DNA methylation can cause heritable phenotypic modifications in the absence of changes in DNA sequence. Environmental stresses can trigger methylation changes and this may have evolutionary consequences, even in the absence of sequence variation. However, it remains largely unknown to what extent environmentally induced methylation changes are transmitted to offspring, and whether observed methylation variation is truly independent or a downstream consequence of genetic variation between individuals. *Genetically identical apomictic dandelion (Taraxacum officinale) plants were exposed to different ecological stresses, and apomictic offspring were raised in a common unstressed environment. We used methylation-sensitive amplified fragment length polymorphism markers to screen genome-wide methylation alterations triggered by stress treatments and to assess the heritability of induced changes. *Various stresses, most notably chemical induction of herbivore and pathogen defenses, triggered considerable methylation variation throughout the genome. Many modifications were faithfully transmitted to offspring. Stresses caused some epigenetic divergence between treatment and controls, but also increased epigenetic variation among plants within treatments. *These results show the following. First, stress-induced methylation changes are common and are mostly heritable. Second, sequence-independent, autonomous methylation variation is readily generated. This highlights the potential of epigenetic inheritance to play an independent role in evolutionary processes, which is superimposed on the system of genetic inheritance. PMID- 20003073 TI - Membrane steroid-binding protein 1 induced by a diffusible fungal signal is critical for mycorrhization in Medicago truncatula. AB - Arbuscular mycorrhiza (AM) is a mutualistic biotrophic association that requires a complex exchange of signals between plant and fungus to allow accommodation of the mycosymbiont in the root cortex. Signal exchange happens even before physical contact, activating the plant symbiotic program. We investigated very early transcriptional responses in Medicago truncatula to inoculation with Glomus intraradices and identified four genes induced by diffusible AM fungal signals before contact. Three of them were previously shown to be mycorrhiza induced at later stages of the symbiosis, while MtMSBP1, encoding a membrane-bound steroid binding protein, is a novel mycorrhizal marker. Expression analyses in plants defective in the symbiotic receptor kinase DMI2 allowed discrimination of two different signaling cascades involved in the perception of the diffusible signals. Thus, while some of the genes are activated in a DMI2-dependent manner, the induction of one of them encoding a proteinase inhibitor is DMI2-independent. Downregulation of MtMSBP1 by RNAi led to an aberrant mycorrhizal phenotype with thick and septated appressoria, decrease number of arbuscules and distorted arbuscule morphology. This provides genetic evidence that MtMSBP1 is critical for mycorrhiza development. We hypothesize that MtMSBP1 plays a role in sterol homeostasis in the root. PMID- 20003074 TI - Regulation of synthesis and degradation of a sulfolipid under sulfur-starved conditions and its physiological significance in Chlamydomonas reinhardtii. AB - Regulation of synthesis and degradation of sulfoquinovosyl diacylglycerol (SQDG), one of the membrane lipids that construct thylakoids, under sulfur (S)-starved conditions and its physiological significance were explored in a green alga, Chlamydomonas reinhardtii. Here, we used sac1 and sac3 mutants defective in response to ambient S-status to characterize the system of known induction of SQDG degradation by S starvation that ensures a major S source for protein synthesis. The SQDG synthesis system was monitored in the wild type during S starvation. An SQDG-deficient mutant, hf-2, was utilized to discover functions where SQDG metabolism participates during S starvation. The induction of SQDG degradation was largely repressed in both sac1 and sac3 mutants. The SQDG synthesis capacity was increased by 40% after S starvation, with a sixfold elevation in the mRNA level of the SQD1 gene for SQDG synthesis. Compared with the wild type, hf-2 had decreased protein accumulation, photosystem (PS) I stability and growth rate. A role of SQDG as an S storage lipid is fulfilled under the control of both SAC1 and SAC3 genes, and it is essential for proper protein synthesis in acclimatization of cells to S starvation. The enhancement in SQDG synthesis may reflect the importance of SQDG as the membrane lipid that stabilizes the PSI complex. PMID- 20003075 TI - Heightened central affective response to visceral sensations of pain and discomfort in IBS. AB - BACKGROUND Typically, conventional functional imaging methods involve repeated exposures to sensory stimulation. In rectal distension (RD) studies that involve multiple distensions, however, it is difficult to disambiguate the central response to RD from pathological alterations in peripheral neural responses associated with relaxation and accommodation of the rectum. METHODS This study addressed potential confounders found in previous imaging studies by collecting functional magnetic resonance imaging studies (fMRI) data during a single slow ramp-tonic distension paradigm and analysing fMRI signal changes using independent component analysis. KEY RESULTS Compared with controls, IBS participants showed increased activation of the anterior cingulate cortices, insula and ventral medial prefrontal regions suggesting heightened affective responses to painful visceral stimuli. In addition, the failure by IBS patients to down-regulate activity within ventral medial prefrontal and the posterior cingulate/precuneus regions was suggestive of reduced sensitivity to somatic changes and delayed shifts away from rest in ;default network' activity patterns. Controls showed heightened activation of the thalamus, striatal regions and dorsolateral prefrontal cortex suggesting greater arousal and salience-driven sustained attention reactions and greater modulation of affective responses to discomfort and pain. CONCLUSION&INFERENCES This work points to alterations in the central response to visceral pain and discomfort in IBS, highlighting diminished modulation and heightened internalization of affective reactions. PMID- 20003076 TI - Glucagon-like peptide-1 inhibits voltage-gated potassium currents in mouse nodose ganglion neurons. AB - BACKGROUND: Glucagon-like peptide-1 (GLP-1) is a major hormone known to regulate glucose homeostasis and gut function, and is an important satiety mediator. These actions are at least in part mediated via an action on vagal afferent neurons. However, the mechanism by which GLP-1 activates vagal afferents remains unknown. We hypothesized that GLP-1 acts on nodose ganglion neuron voltage-gated potassium (KV) channels, increasing membrane excitability. METHODS: Employing perforated patch clamp recordings we examined the effects of GLP-1 on membrane properties as well as voltage-gated potassium currents. Extracellular recordings of jejunal afferents were performed to demonstrate the functional relevance of these effects at the nerve terminal. KEY RESULTS: Glucagon-like peptide-1 depolarized a subpopulation of nodose neurons. This membrane depolarization was used to identify neurons containing functional GLP-1 receptors. In these neurons, GLP-1 decreased rheobase and broadened the action potential, and increased the number of action potentials elicited at twice rheobase. We identified a GLP-1 sensitive current whose reversal potential shifted in a depolarizing direction when extracellular potassium was increased. We identified two macroscopic K currents, IA, an inactivating current and IK a sustained current. GLP-1 caused inhibition of these currents, IK by 45%, P < 0.05 and IA currents by 52%P < 0.01, associated with a hyperpolarizing shift of steady-state inactivation curves for both currents. In extracellular recordings of jejunal afferents, GLP-1 increased firing rate, the effect blocked by the K(+) channel antagonist 4-AP. CONCLUSIONS & INFERENCES: These experiments indicate that GLP-1 receptor activation results in vagal afferent excitation, due at least in part to inhibition of sustained and inactivating potassium currents. This mechanism may be important in satiety and glucose homeostatic signals arising from the gastrointestinal tract. PMID- 20003078 TI - Inhibition of p38 MAPK improves intestinal disturbances and oxidative stress induced in a rabbit endotoxemia model. AB - BACKGROUND: Lipopolysaccharide (LPS) decreases intestinal contractility and induces the release of reactive oxygen species, which play an important role in the pathogenesis of sepsis. p38 mitogen-activated protein kinase (MAPK) can be activated by a variety of stimuli such as LPS. The aims of this study were: (i) to investigate the role of p38 MAPK in the effect of LPS on (a) the acetylcholine, prostaglandin E(2) and KCl-induced contractions of rabbit duodenum and (b) the oxidative stress status; (ii) to localize the active form of p38 in the intestine. METHODS: Rabbits were injected with (i) saline, (ii) LPS, (iii) SB203580, a specific p38 MAPK inhibitor or (iv) SB203580 + LPS. Duodenal contractility was studied in an organ bath. SB203580 was also tested in vitro. The protein expression of p-p38 and total p38 was measured by Western blot and p p38 was localized by immunohistochemistry. The formation of products of oxidative damage to proteins (carbonyls) and lipids (MDA+4-HDA) was quantified in intestine and plasma. KEY RESULTS: ACh, PGE(2) and KCl-induced contractions decreased with LPS. LPS increased phospho-p38 expression and the levels of carbonyls and MDA+4 HDA. SB203580 blocked the effect of LPS on the ACh, PGE(2) and KCl-induced contractions in vivo and in vitro and the levels of carbonyls and MDA+4-HDA. P p38 was detected in neurons of the myenteric plexus and smooth muscle cells of duodenum. CONCLUSIONS & INFERENCES: Lipopolysaccharide decreases the duodenal contractility in rabbits and increases the production of free radicals. p38 MAPK is a mediator of these effects. PMID- 20003079 TI - 5-HT(2B) receptors modulate visceral hypersensitivity in a stress-sensitive animal model of brain-gut axis dysfunction. AB - BACKGROUND: Irritable bowel syndrome (IBS) is associated with an enhanced perception to visceral stimuli and exaggerated stress response. The serotonergic neurotransmitter system has been strongly implicated as a key player in the manifestation of IBS symptomatology including visceral hypersensitivity. However the role of 5-HT(2B) receptors in visceral pain, although speculated, is currently unclear. Thus we assessed the impact of a selective 5-HT(2B) receptor antagonist, RS-127445, on visceral hypersensitivity in a model of brain gut axis dysfunction the Wistar Kyoto (WKY) rat. METHODS: Colorectal distension (CRD) was used to assess the visceral sensitivity of the WKY rat compared to normosensitive Sprague Dawley (SD) rats. Once we verified the visceral sensitivity of the WKY rat we assessed the efficacy of RS-127445 in pain signalling from the colorectum. We administered the compound peripherally (i.p.) and centrally (i.c.v.) in order to ascertain the site of action of RS 127445. Behavioural responses to colorectal distention were then monitored. KEY RESULTS: The WKY rats were more viscerally hypersensitive than the SD as previously shown. RS-127445 (5 mg kg(-1), i.p.) significantly reversed visceral hypersensitivity in WKY animals. Moreover, when administered intracerebroventricularly RS-127445 (100 nM) also decreased the number of pain behaviours during noxious CRD in the WKY animals. CONCLUSIONS & INFERENCES: Taken together, blockade of 5-HT(2B) receptors offers an exciting novel therapeutic target for pain relief in stress-related gastrointestinal disorders such as IBS. PMID- 20003080 TI - Cognitive symptoms, cervical range of motion and pain as prognostic factors after whiplash trauma. AB - OBJECTIVES: To evaluate pain, cervical range of motion (CROM) and cognitive symptoms as predictors for poor prognosis defined as sick leave 3 years later. MATERIAL AND METHODS: In 97 patients CROM, pain intensity and cognitive symptoms were measured immediately following trauma, at 6 months and 3 years. Patients were also asked at 3 years if they had been on sick leave the last 6 months. RESULTS: Pain intensity and reduced CROM were not clinically useful as predictors of later sick leave. The best predictors were presence within 96 h after injury of the two cognitive symptoms 'being easily distracted' (odds ratio 8.7-50) and 'easily irritated' (odds ratio 5.3-31). CONCLUSIONS: Initial pain and reduced CROM may be related to minor tissue damage which often heals while late functionality is more dependent on other factors such as cognitive dysfunction. For patients with whiplash-associated disorders two simple questions should be asked; 'Are you currently easily irritated?' and 'Are you currently easily distracted (e.g. is it difficult for you to follow a conversation if several people are talking in the room at the same time)?'. An affirmative answer to any of these questions indicates an increased risk for poor prognosis defined as sick leave 3 years later. PMID- 20003077 TI - Gastroparesis and functional dyspepsia: excerpts from the AGA/ANMS meeting. AB - BACKGROUND: Despite the relatively high prevalence of gastroparesis and functional dyspepsia, the aetiology and pathophysiology of these disorders remain incompletely understood. Similarly, the diagnostic and treatment options for these two disorders are relatively limited despite recent advances in our understanding of both disorders. PURPOSE: This manuscript reviews the advances in the understanding of the epidemiology, pathophysiology, diagnosis, and treatment of gastroparesis and functional dyspepsia as discussed at a recent conference sponsored by the American Gastroenterological Association (AGA) and the American Neurogastroenterology and Motility Society (ANMS). Particular focus is placed on discussing unmet needs and areas for future research. PMID- 20003081 TI - Long-term levetiracetam treatment in patients with epilepsy: 3-year follow up. AB - OBJECTIVES: To assess the long-term efficacy and tolerability of levetiracetam in routine clinical practice. MATERIALS AND METHODS: We retrospectively analysed 218 patients, mostly adults, presenting mostly with localisation-related epilepsy, treated with levetiracetam as adjunctive therapy or monotherapy for up to 36 months. The primary points evaluated were: long-term retention rate, reasons for discontinuing levetiracetam and the percentage of seizure-free patients. RESULTS: The retention rate at 6, 12, 24 and 36 months following the commencement of levetiracetam treatment was 91.7, 75.2, 60.1 and 53.7% respectively. Sixty-seven (30.7%) patients discontinued levetiracetam treatment. During the clinical audit evaluation period, surgical resection or implantation of VNS was performed in 31 (14.3%) patients. In 53 of the 67 patients (79.1%), the treatment was discontinued due to lack of efficacy; in 14 patients (20.9%) treatment was discontinued due to adverse events. In total, 24 of 218 patients (11.0%) were seizure-free for 36 months. CONCLUSIONS: Levetiracetam is an effective and well tolerated option for long-term treatment of epilepsy in adults. PMID- 20003082 TI - Validation of Serbian version of the disease-specific myasthenia gravis questionnaire. AB - AIM: The aim of this study was to validate translated and cross-cultural adapted Italian version of myasthenia gravis-specific questionnaire (MGQ) in Serbian MG patients. MATERIALS AND METHODS: The questionnaire was validated in 140 consecutive MG patients from Belgrade. In each patient association between the total MGQ score and form and severity of the disease was determined. Also, correlation between regional domain scores of MGQ and main clinical findings according to Besinger's clinical score was analyzed. RESULTS: Patients' participation in the assessment was satisfactory with excellent internal consistency and reproducibility. Total MGQ score, as well as domain scores, correlated with highly significant inverse relationship with the disease severity and clinical status of patients at the moment of completing the questionnaire. Furthermore, the bulbar domain of the questionnaire appeared more specific and sensitive than clinical history and examination. CONCLUSION: We concluded that the Serbian version of the MGQ may be useful as a measure of clinical outcome in patients with MG. PMID- 20003083 TI - Polyneuropathy associated with IgM vs IgG monoclonal gammopathy: comparison between clinical and electrophysiological findings. AB - OBJECTIVE: The neuropathy associated with IgM monoclonal gammopathy (IgM-MG) is regarded as a sensorimotor, mainly demyelinating neuropathy. It is not fully known whether the neuropathy in IgG-MG is caused by the same mechanisms and shows the same electrophysiological characteristics. We aimed at making a comparison between clinical and neurophysiological findings in these two conditions. PATIENTS AND METHODS: Twenty-seven patients with IgM-associated neuropathy [18 with anti-myelin-associated glycoprotein (anti-MAG) antibodies] were compared with 15 age-matched patients with IgG-associated neuropathy. RESULTS: Patients with IgM-associated neuropathy (especially those with anti-MAG antibodies) had significantly clinically more severe disabilities with involvement of both motor and sensory functions compared with patients with IgG-associated neuropathy in whom clinical sensory disturbances were more prominent than motor dysfunction. Motor and sensory conduction velocities were significantly lower and distal latencies significantly longer in the IgM group than in the IgG group concerning the median, ulnar and peroneal nerves. Fifty-four per cent of the patients in the IgM group did not present a sensory response of the median nerve vs 13% in the IgG group. There was also a significant difference concerning absent responses from the peroneal and sural nerves in the IgM vs IgG group (peroneal: 48% vs 13%, sural: 88% vs 27%). CONCLUSION: Polyneuropathy associated with IgM-MG, especially when associated with anti-MAG antibodies, appears to have more of a demyelinating involvement that meets the criteria for demyelination. This was not as clear in those associated with IgG. The IgG neuropathy showed less and milder deficit in the electrophysiological studies. PMID- 20003084 TI - Motor impairment in liver cirrhosis without and with minimal hepatic encephalopathy. AB - AIM: Manifest hepatic encephalopathy (HE) goes along with motor symptoms such as ataxia, mini-asterixis, and asterixis. The relevance of motor impairments in cirrhotics without and with minimal HE (mHE) is still a matter of debate. PATIENTS AND METHODS: We tested three different groups of patients with liver cirrhosis: no signs of HE (HE 0), mHE, and manifest HE grade 1 according to the West Haven criteria (HE 1). All patients (n = 24) and 11 healthy control subjects were neuropsychometrically tested including critical flicker frequency (CFF), a reliable measure for HE. Motor abilities were assessed using Fahn Tremor Scale and International Ataxia Rating Scale. Fastest alternating index finger movements were analyzed for frequency and amplitude. RESULTS: Statistical analyses showed an effect of HE grade on tremor and ataxia (P < 0.01). Additionally, both ratings yielded strong negative correlation with CFF (P < 0.01, R = -0.5). Analysis of finger movements revealed an effect of HE grade on movement frequency (P < 0.03). Moreover, decreasing movement frequency and increasing movement amplitude parallel decreasing CFF (P < 0.01, R = 0.6). CONCLUSION: Our results indicate that ataxia, tremor, and slowing of finger movements are early markers for cerebral dysfunction in HE patients even prior to neuropsychometric alterations becoming detectable. PMID- 20003085 TI - Chronic inflammatory demyelinating polyradiculoneuropathy in children: characterized by subacute, predominantly motor dominant polyeuropathy with a favorable response to the treatment. AB - OBJECTIVES: Chronic inflammatory demyelinating polyradiculopathy (CIDP) is less well-studied in children than in adults, probably due to its relative rarity. This study was performed in order to characterize the clinical features of CIDP in children. MATERIALS AND METHODS: Twenty-eight patients with CIDP who were followed up for more than 1 year were included, and were divided into a child (n = 7, age <16) and an adult group (n = 21, age >or=16). Then, we have assessed the initial progression pattern, clinical course, and serial nerve conduction findings in each patient. Finally, differential features in child and adult group were analyzed. RESULTS: Distinguishing features in the child group include subacute progression within less than 2 months, predominant motor system involvement in lower extremities, and marked improvement in response to immune modulating therapy. Our study also suggested that serial nerve conduction study may be useful in assessing the effectiveness of the treatment in children. CONCLUSIONS: Our study showed that children with CIDP have some distinguishing features from adults in terms of clinical course and response to treatment. PMID- 20003086 TI - Misdiagnosis of epileptic and non-epileptic seizures in a neurological intensive care unit. AB - OBJECTIVE: The etiological misinterpretation of paroxysmal neurological symptoms frequently causes a delayed treatment or an inappropriate utilization of ICU capacities. METHODS: In this study, the data of 208 patients admitted to a neurological ICU because of acute transient neurological deficits, loss of consciousness or unclear motor phenomena were retrospectively analyzed. The initial emergency room diagnosis was compared to the final diagnosis and the rate of misdiagnosis was related to the patients' history and diagnostic data. RESULTS: In 13.9%, the emergency room diagnosis of epileptic seizures turned out to be incorrect, whereas in 15.6%, the final diagnosis of epileptic seizures was missed in the emergency room. Factors that were significantly correlated to missing the seizure diagnosis were (i) no prior history of epilepsy, (ii) old age, (iii) multi-morbidity, (iv) pathologic CT-scans demonstrating cerebrovascular lesions, (v) seizure description by non-professionals, (vi) predominantly negative seizure phenomena (aphasia, loss of consciousness, paresis), (vii) lack of tongue-bite lesions. PMID- 20003087 TI - Circulating free nitrotyrosine and cognitive decline. AB - OBJECTIVE: To determine if the circulating nitrotyrosine level significantly correlates with parameters measuring cognitive abilities. MATERIALS AND METHODS: One-hundred and twelve community-living subjects (ranging in age from 27 to 98 years) were evaluated for cognitive abilities [Mini Mental State Examination (MMSE) score] and circulating free nitrotyrosine plasma level, as well as for several variables that might influence cognitive abilities (age, education) and nitrotyrosine level (body mass index, haematological parameters, cardiovascular and inflammatory indices). RESULTS: In the sub-group of cognitively impaired subjects (score at MMSE < 23.9), but not in that of cognitively not impaired subjects, a significant inverse correlation exists between nitrotyrosine level and MMSE score (r = -0.378; P < 0.02). CONCLUSIONS: The finding, if confirmed by longitudinal studies, could play a role in the management of the subjects with Mild Cognitive Impairment, the clinical condition considered as a transitional state between the changes of cognitive ability in normal aging and dementia. PMID- 20003088 TI - Modulation of seizure threshold by vagus nerve stimulation in an animal model for motor seizures. AB - OBJECTIVE: The precise mechanism of action of vagus nerve stimulation (VNS) in suppressing epileptic seizures remains to be elucidated. This study investigates whether VNS modulates cortical excitability by determining the threshold for provoking focal motor seizures by cortical electrical stimulation before and after VNS. MATERIAL AND METHODS: Male Wistar rats (n = 8) were implanted with a cuff-electrode around the left vagus nerve and with stimulation electrodes placed bilaterally on the rat motor cortex. Motor seizure threshold (MST) was assessed for each rat before and immediately after 1 h of VNS with standard stimulation parameters, during two to three sessions on different days. RESULTS: An overall significant increase of the MST was observed following 1 h of VNS compared to the baseline value (1420 microA and 1072 microA, respectively; P < 0.01). The effect was reproducible over time with an increase in MST in each experimental session. CONCLUSIONS: VNS significantly increases the MST in a cortical stimulation model for motor seizures. These data indicate that VNS is capable of modulating cortical excitability. PMID- 20003089 TI - Black holes in multiple sclerosis: definition, evolution, and clinical correlations. AB - Magnetic resonance imaging (MRI) is a sensitive paraclinical test for diagnosis and assessment of disease progression in multiple sclerosis (MS) and is often used to evaluate therapeutic efficacy. The formation of new T2-hyperintense MRI lesions is commonly used to measure disease activity, but lacks specificity because edema, inflammation, gliosis, and axonal loss all contribute to T2 lesion formation. As the role of neurodegeneration in the pathophysiology of MS has become more prominent, the formation and evolution of chronic or persistent Tl hypointense lesions (black holes) have been used as markers of axonal loss and neuronal destruction to measure disease activity. Despite the use of various detection methods, including advanced imaging techniques such as magnetization transfer imaging and magnetic resonance spectroscopy, correlation of persistent black holes with clinical outcomes in patients with MS remains uncertain. Furthermore, although axonal loss and neuronal tissue destruction are known to contribute to irreversible disability in patients with MS, there are limited data on the effect of therapy on longitudinal change in Tl-hypointense lesion volume. Measurement of black holes in clinical studies may elucidate the underlying pathophysiology of MS and may be an additional method of evaluating therapeutic efficacy. PMID- 20003090 TI - Subjective memory decline in healthy community-dwelling elders. What does this complain mean? AB - OBJECTIVES: Subjective feelings of memory decline are fairly common among the elderly. The causes of this are heterogeneous, and may be related to both affective and cognitive disorders. We attempted to explore the associations between subjective and cognitive measures. MATERIALS AND METHODS: Healthy subjects were studied. They completed questionnaires regarding memory difficulties and lifestyle habits, the Geriatric Depression scale (GDS), and the Spielberger State-Trait Anxiety Inventory. Cognitive functions were tested using the Mini-Mental State Exam and supplemented with NeuroTrax, a computerized neurophysiological battery. Univariate logistic regression model was applied to estimate odd ratios (OR) and 95% confidence intervals of associations. RESULTS: Of 341 consecutive non-depressed subjects, 257 participants (75.4%) reported subjective memory decline (SMD). Subjects with and without SMD did not differ in age, gender, education, marital status, employment and life-style. Subjects with SMD had elevated GDS scores (OR = 1.14, 95% CI: 1.003-1.29), white anxiety level showed a tendency to be increased (OR = 1.03, 95% CI: 0.99-1.06). Comparison of cognitive performance has not revealed differences in cognitive domains between subjects with and without SMD. CONCLUSIONS: SMD in healthy elderly people is associated with sub-clinical depression even among those without objectively measured cognitive decline. PMID- 20003091 TI - Seasonal allergies and suicidality: results from the National Comorbidity Survey Replication. AB - OBJECTIVE: Studies have shown an association between allergies and suicidality, and a seasonality of suicide has also been described. We hypothesize an association between history of seasonal allergies and suicide ideation and attempt. METHOD: Data came from the National Comorbidity Survey Replication, a nationally representative sample (n = 5692) of adults living in the US. Logistic regression models were used to calculate adjusted odds ratios (OR) controlling for the following: age, sex, race, smoking, asthma and depression. RESULTS: After weighting and adjustment, a positive and statistically significant association was found between history of seasonal allergies and history of suicidal ideation [adjusted OR = 1.27 (1.01-1.58)]. We found no association between history of seasonal allergies and history of suicide attempts [adjusted OR = 1.17 (0.89 1.52)]. CONCLUSION: Findings from a population-based sample support the hypothesized relationship between allergies and suicidal ideation. PMID- 20003092 TI - Prevalence and predictors of recurrence of major depressive disorder in the adult population. AB - OBJECTIVE: Knowledge of the risk of recurrence after recovery of a major depressive disorder (MDD) is of clinical and scientific importance. The purpose of this paper was to provide a systematic review of the prevalence and predictors of recurrence of MDD. METHOD: Studies were searched in Medline en PsychINFO using the search terms 'recur*', 'relaps*', 'depress*', 'predict*' and course. RESULTS: Recurrence of MDD in specialised mental healthcare settings is high (60% after 5 years, 67% after 10 years and 85% after 15 years) and seems lower in the general population (35% after 15 years). Number of previous episodes and subclinical residual symptoms appear to be the most important predictors. Gender, civil status and socioeconomic status seem not related to the recurrence of MDD. CONCLUSION: Clinical factors seem the most important predictors of recurrence. Data from studies performed in the general population and primary care on the recurrent course of MDD are scarce. PMID- 20003093 TI - The clinical presentation and prognostic factors for intrahepatic and extrahepatic cholangiocarcinoma in a tertiary care centre. AB - BACKGROUND: The incidence of cholangiocarcinoma is rising. Accurate predictors of survival at diagnosis are not well defined. AIM: To clarify the clinical presentation and prognostic factors of intrahepatic cholangiocarcinoma and extrahepatic cholangiocarcinoma in a contemporary cohort of patients. METHODS: Records for consecutive patients at the University of Michigan hospital diagnosed with cholangiocarcinoma between January 2003 and April 2008 were reviewed. RESULTS: In all, 136 patients had cholangiocarcinoma (79 intra- and 57 extrahepatic cholangiocarcinoma). Median survival was 27.3 months-25.8 months for intrahepatic cholangiocarcinoma and 30.3 months for extrahepatic cholangiocarcinoma. Independent predictors of mortality at presentation on multivariate analysis were elevated bilirubin level (HR 1.04, 95%CI 1.01-1.07), CA 19-9 levels >100 U/mL (HR 1.90, 95%CI 1.17-3.08) and stage of disease (HR 1.51, 95%CI 1.16-1.96). After adjusting for baseline prognostic factors, surgical therapy was associated with improved survival (HR 0.48; 95% CI 0.26-0.88). There were no significant differences regarding clinical presentation, disease stage (P = 0.98), and survival (P = 0.51) between intra- and extrahepatic cholangiocarcinoma. CONCLUSIONS: Survival for cholangiocarcinoma remains poor with no significant difference in outcomes between intra- and extrahepatic cholangiocarcinoma. Stage of disease, bilirubin level and CA 19-9 level are important prognostic factors at presentation. Surgical therapy provides similar efficacy for both tumours when adjusted for other prognostic variables. PMID- 20003094 TI - A pneumatic dilation strategy in achalasia: prospective outcome and effects on oesophageal motor function in the long term. AB - BACKGROUND: Long-term follow-up studies of achalasia after pneumatic dilation, mostly retrospective, have shown variable results. AIM: To examine the outcome of achalasia after pneumatic dilation using a prospective follow-up programme. METHODS: One or two dilations (first dilation treatment) in 77 patients to achieve stable (>1 year) remission and patients followed up with yearly clinical and manometric assessments. Endoscopy, pH monitoring and barium swallow were also performed. RESULTS: A total of 69 patients achieved stable remission and were followed up for 5.6 years (3-10.7) [median (IQ range)], whereas six patients underwent cardiomyotomy and two experienced a perforation. Twelve of the 69 patients relapsed after 2.6 years (1.7-5.1): nine of 12 underwent one to two further dilations. Six-year remission rate (by Kaplan-Meyer estimates) was 82% after first dilation treatment and 96% after all dilations. Continuous antisecretory treatment was clinically needed in 16%, oesophagitis present in 7% and reflux pathological in 28% of the patients. Beneficial effects of dilation on oesophageal motility and on diameter of the oesophageal body at barium swallow were maintained during follow-up. CONCLUSIONS: A management strategy including sessions of pneumatic dilation until stable remission and a standardized follow up is highly successful in the long term. Gastro-oesophageal reflux is clinically relevant in a minority of patients. PMID- 20003095 TI - Clinical trial: the efficacy of alverine citrate/simeticone combination on abdominal pain/discomfort in irritable bowel syndrome--a randomized, double blind, placebo-controlled study. AB - BACKGROUND: Alverine citrate and simeticone combination has been used for almost 20 years in irritable bowel syndrome (IBS), but supportive scientific evidence of efficacy was limited. AIM: To evaluate the efficacy of alverine citrate and simeticone combination in patients with IBS-related abdominal pain/discomfort. METHODS: A total of 412 IBS patients meeting ROME III criteria were included in this double-blind randomized placebo-controlled study if their abdominal pain/discomfort intensity was at least 60 mm on a 0-100 mm visual analogue scale (VAS) during a 2-week run-in treatment-free period. Patients were randomly assigned through the use of Interactive Voice Response System to receive either alverine citrate 60 mg with simeticone 300 mg three times daily or matching placebo for 4 weeks. RESULTS: The full analysis set included 409 patients (71.4% female: mean age: 46.2 +/- 13.9 years). At week 4, alverine citrate and simeticone group had lower VAS scores of abdominal pain/discomfort (median: 40 mm vs. 50 mm, P = 0.047) and higher responder rate (46.8% vs. 34.3%, OR = 1.3; P = 0.01) as compared with placebo group. Patient receiving alverine citrate and simeticone reported greater global symptom improvement compared with those receiving placebo (P = 0.0001). Reported adverse events were similar in both groups. CONCLUSION: Alverine citrate/simeticone combination was significantly more effective than placebo in relieving abdominal pain/discomfort in patients with IBS. PMID- 20003096 TI - Predicting the advent of ascites and other complications in primary biliary cirrhosis: a staged model approach. AB - BACKGROUND: Current survival models for primary biliary cirrhosis have limited precision for medium and long-term survival. Aim To describe a prognostic model for the advent of complications in primary biliary cirrhosis as the first approach to a staged prognostic model. METHODS: From an established database of 289 consecutive primary biliary cirrhosis patients referred to Royal Free Hospital over 12 years (mean follow-up of 4.1 years), baseline characteristics at referral were evaluated by Cox-proportional hazards regression modelling. RESULTS: The following complications occurred de novo: 85 ascites/peripheral oedema, 40 oesophagogastric varices, 63 encephalopathy, 29 spontaneous bacterial peritonitis and/or septicaemia, 59 symptomatic urinary tract infections. Age, albumin, log(10)(bilirubin), presence of ascites at referral, variceal bleeding within 6 weeks before referral, detection of oesophagogastric varices at or before referral were significant at multivariate analysis with different combinations and coefficients for each complication. The model for predicting ascites and/or peripheral oedema best fitted the observed data (ROC = 0.7682, S.E. = 0.0385). CONCLUSIONS: The known prognostic factors in primary biliary cirrhosis also model the advent of complications. In view of the prognostic importance of ascites and its more robust statistical model, ascites and/or peripheral oedema could represent, following validation, the most suitable staged model in primary biliary cirrhosis to improve precision in survival modelling. PMID- 20003097 TI - Role of ataxia telangiectasia mutated in insulin signalling of muscle-derived cell lines and mouse soleus. AB - AIM: Ataxia telangiectasia mutated (ATM) reportedly plays a role in insulin stimulated activation of Akt in some cell types but not in others. The role of ATM in insulin signalling has not been firmly resolved for skeletal muscle cells, for which Akt phosphorylation is a pivotal step in stimulation of glucose transport. Accordingly, our aim was to determine the role of ATM in insulin effects for cell lines derived from skeletal muscle and for skeletal muscle. METHODS: We examined insulin effects in L6 myotubes, mouse soleus, C2C12 myotubes and differentiated rhabdomyosarcoma (RD) cells in the presence and absence of a low concentration (1 microm) of the ATM inhibitor KU55933. We also compared insulin signalling in C2C12 cells expressing shRNA against ATM and control cell lines (empty vector; cells expressing non-targeting shRNA). RESULTS: In L6 myotubes and mouse soleus muscle, KU55933 inhibited insulin-stimulated phosphorylation of the 160 kDa substrate of Akt (AS160) despite no effect on Akt. In contrast, KU55933 prevented insulin-stimulated Akt phosphorylation in C2C12 myotubes. Furthermore, C2C12 myotubes expressing shRNA against ATM displayed reduced insulin-stimulated Akt phosphorylation compared to controls. KU55933 also decreased insulin-stimulated Akt phosphorylation in differentiated RD cells. CONCLUSION: These model-dependent differences in the role of ATM in insulin action demonstrate a role of ATM in insulin-stimulated phosphorylation of Akt (in C2C12 and RD cells) but also allow the elucidation of a novel, Akt-independent role of ATM (in L6 myotubes and mouse soleus, at the level of AS160) in insulin signalling. PMID- 20003098 TI - Characterization of sleep-wake patterns in a novel transgenic mouse line overexpressing human prepro-orexin/hypocretin. AB - AIM: Orexin/hypocretin peptides are expressed in the lateral hypothalamus and involved in the regulation of autonomic functions, energy homeostasis and arousal states. The sleep disorder narcolepsy, which is characterized by excessive daytime sleepiness and occurrence of sudden rapid eye movement (REM) sleep, is associated with a loss of orexin neurones. Our study investigated the effects of orexins on sleep-wake patterns in a novel transgenic mouse line overexpressing the human prepro-orexin (hPPO) gene under the control of its endogenous promoter. METHODS: Orexin overexpression was investigated by PCR, Southern and Western blotting as well as immunohistochemistry. Polysomnographic recordings were performed for analyses of sleep-wake patterns and for electroencephalographic activity during 24 h baseline and during and after 6 h of sleep deprivation (SD). RESULTS: Transgenic hPPO mice had increased expression of human prepro-orexin (hPPO) and orexin-A in the hypothalamus. Transgene expression decreased endogenous orexin-2 receptors but not orexin-1 receptors in the hypothalamus without affecting orexin receptor levels in the basal forebrain, cortex or hippocampus. Transgenic mice compared with their wild type littermates showed small but significant differences in the amount of waking and slow wave sleep, particularly during the light-dark transition periods, in addition to a slight reduction in REM sleep during baseline and during recovery sleep after SD. CONCLUSION: The hPPO-overexpressing mice show a small reduction in REM sleep, in addition to differences in vigilance state amounts in the light/dark transition periods, but overall the sleep-wake patterns of hPPO-overexpressing mice do not significantly differ from their wild type littermates. PMID- 20003099 TI - Effects of short-term food deprivation on orexin-A-induced intestinal bicarbonate secretion in comparison with related secretagogues. AB - Studies of gastrointestinal physiology in humans and intact animals are usually conducted after overnight fast. We compared the effects of orexin-A, vasoactive intestinal polypeptide (VIP), melatonin, serotonin, uroguanylin, ghrelin and prostaglandin E(2) (PGE(2)) on duodenal bicarbonate secretion in fed and overnight fasted animals. This review is a summary of our findings. Secretagogues were administered by intra-arterial infusion or luminally (PGE(2)). Enterocyte intracellular calcium ([Ca(2+)](i)) signalling was studied by fluorescence imaging. Total RNA was extracted, reverse transcripted to cDNA and expression of orexin receptors measured by quantitative real-time PCR. Orexin-A stimulates the duodenal secretion in continuously fed animals but not in food-deprived animals. Similarly, short-term fasting causes a 100-fold decrease in the amount of the muscarinic agonist bethanechol required for stimulation of secretion. In contrast, fasting does not affect secretory responses to intra-arterial VIP, melatonin, serotonin, uroguanylin and ghrelin, or that to luminal PGE(2). Orexin A induces [Ca(2+)](i) signalling in enterocytes from fed rats but no significant [Ca(2+)](i) responses occur in enterocytes from fasted animals. In addition, overnight fasting decreases the expression of mucosal orexin receptors. Short term food deprivation thus decreases duodenal expression of orexin receptors and abolishes the secretory response to orexin-A as well as orexin-A-induced [Ca(2+)](i) signalling. Fasting, furthermore, decreases mucosal sensitivity to bethanechol. The absence of declines in secretory responses to other secretagogues tested strongly suggests that short-term fasting does not affect the secretory capacity of the duodenal mucosa in general. Studies of intestinal secretion require particular evaluation with respect to feeding status. PMID- 20003100 TI - Probing the corticospinal link between the motor cortex and motoneurones: some neglected aspects of human motor cortical function. AB - This review considers the operation of the corticospinal system in primates. There is a relatively widespread cortical area containing corticospinal outputs to a single muscle and thus a motoneurone pool receives corticospinal input from a wide region of the cortex. In addition, corticospinal cells themselves have divergent intraspinal branches which innervate more than one motoneuronal pool but the synergistic couplings involving the many hand muscles are likely to be more diverse than can be accommodated simply by fixed patterns of corticospinal divergence. Many studies using transcranial magnetic stimulation of the human motor cortex have highlighted the capacity of the cortex to modify its apparent excitability in response to altered afferent inputs, training and various pathologies. Studies using cortical stimulation at 'very low' intensities which elicit only short-latency suppression of the discharge of motor units have revealed that the rapidly conducting corticospinal axons (stimulated at higher intensities) drive motoneurones in normal voluntary contractions. There are also major non-linearities generated at a spinal level in the relation between corticospinal output and the output from the motoneurone pool. For example, recent studies have revealed that the efficacy of the human corticospinal connection with motoneurones undergoes activity-dependent changes which influence the size of voluntary contractions. Hence, corticospinal drives must be sculpted continuously to compensate for the changing functional efficacy of the descending systems which activate the motoneurones. This highlights the need for proprioceptive monitoring of movements to ensure their accurate execution. PMID- 20003101 TI - Pain in children with cerebral palsy: a cross-sectional multicentre European study. AB - AIM: To determine the prevalence and associations of self-reported and parent reported pain in children with cerebral palsy (CP) of all severities. METHOD: Cross-sectional design using a questionnaire; analysis using ordinal regression. Children aged 8-12 years were randomly selected from population-based registers of children with CP in eight European regions; a further region recruited 75 children from multiple sources. Outcome measures were pain in the previous week among children who could self-report and parents' perception of their child's pain in the previous 4 weeks. RESULTS: Data on pain were available from 490 children who could self-report and parents of 806 children (those who could and could not self-report). The estimated population prevalence of self-reported pain in the previous week was 60% (95% CI: 54-65%) and that of parent-reported pain in the previous 4 weeks was 73% (95% CI: 69-76%). In self-reporting children, older children reported more pain but pain was not significantly associated with severity of impairment. In parent reports, severity of child impairment, seizures and parental unemployment were associated with more frequent and severe pain. CONCLUSION: Pain in children with CP is common. Clinicians should enquire about pain and consider appropriate physical, therapeutic or psychological management. PMID- 20003102 TI - The effect of ascorbate on minor recurrent aphthous stomatitis. AB - AIM: Minor recurrent aphthous stomatitis (MRAS) is a common, painful and inflammatory ailment of the oral cavity with juvenile onset and unknown aetiology. The purpose of this study was to evaluate the potential of ascorbate (vitamin C) to reduce the frequency of MRAS and severity of pain. PATIENTS AND METHODS: Sixteen MRAS patients (9 boys and 7 girls: mean age, 12.0 +/- 2.4 years old) were assigned to take an oral dosage of 2000 mg/m(2)/day ascorbate. SUBJECTS: Their baseline frequency of outbreaks and the level of pains were compared during the treatment; in addition, a crossover clinical trial was performed. Polymorphonuclear leucocytes play a role in the pathogenesis, and then superoxide anion production was evaluated in prior to ascorbate treatment. RESULTS: The data indicated a statistically significant 50% reduction in oral ulcer outbreaks and a decline of pain level. Neutrophils were primed for superoxide anion production in the patients with MRAS. CONCLUSION: Ascorbate may modulate the generation of reactive oxygen species and augment neutrophil apoptosis, which could prevent neutrophil-mediated inflammation. Ascorbate seems to be effective, but the findings of our study were preliminary and it should be re-evaluated with a larger randomized controlled clinical trials. PMID- 20003103 TI - Thimerosal exposure (from tetanus-diphtheria vaccine) during pregnancy and neurodevelopment of breastfed infants at 6 months. AB - AIM: We studied the effect on neurodevelopment of infants who are exposed to thimerosal in tetanus-diphtheria (Td) vaccines during pregnancy. METHODS: We compared Gesell Developmental Schedules (GDS) of exclusive breastfed infants at 6 months born to mothers who received Td (1 to 3 doses) against those who were born to mothers who did not take such vaccines. RESULTS: Compared with the group of infants not exposed to ethylmercury in utero, the infants of exposed mothers showed no significant difference in neurodevelopment delays. Although there was a significant correlation between hair-Hg of mothers and hair-Hg of neonates (Spearman r = 0.353; p = 0.0011), there was no significant correlation between the level of in utero exposure to ethylmercury in Td vaccines and neonate's hair Hg concentrations (Spearman r = 0.060; p = 0.5922). However, regression analysis showed that GDS at 6 months was significantly associated with total mercury concentration of neonate's hair but was not sensitive to the number of vaccines taken by the mother. CONCLUSION: Early neurodevelopment of exclusively breastfed infants is sensitive to in utero exposure to mercury, but maternal thimerosal exposure in tetanus-diphtheria vaccines per se cannot portend clinical neurodevelopment delays measured by GDS at 6 months. PMID- 20003104 TI - Down syndrome and streptococcus group A disease in hospitalized children. PMID- 20003105 TI - Respiratory viral infection in lower airways of asymptomatic children. AB - AIM: The aim of this study was to determine if asthmatic children have viruses more commonly detected in lower airways during asymptomatic periods than normal children. METHODS: Fifty-five asymptomatic children attending elective surgical procedures (14 with stable asthma, 41 normal controls) underwent non bronchoscopic bronchoalveolar lavage. Differential cell count and PCR for 13 common viruses were performed. RESULTS: Nineteen (35%) children were positive for at least one virus, with adenovirus being most common. No differences in the proportion of viruses detected were seen between asthmatic and normal 'control' children. Viruses other than adenovirus were associated with higher neutrophil counts, suggesting that they caused an inflammatory response in both asthmatics and controls (median BAL neutrophil count, 6.9% for virus detected vs. 1.5% for virus not detected, p = 0.03). CONCLUSIONS: Over one-third of asymptomatic children have a detectable virus (most commonly adenovirus) in the lower airway; however, this was not more common in asthmatics. Viruses other than adenovirus were associated with elevated neutrophils suggesting that viral infection can be present during relatively asymptomatic periods in asthmatic children. PMID- 20003106 TI - Band injury caused by cord around leg in utero. PMID- 20003107 TI - A four-year prospective study on intraocular pressure in relation to phacoemulsification cataract surgery. AB - AIM OF STUDY: To follow up prospectively the intraocular pressure (IOP) of healthy eyes with senile cataract undergoing phacoemulsification surgery over a duration of 4 years. PATIENTS AND METHODS: Thirty-five patients entering first eye cataract surgery had IOP measured by applanation tonometry pre-operatively and on day 1, at 1 month, 6 months, 1 year, 2 years and 4 years after surgery at 9 a.m. and again at 2 p.m. in the Department of Ophthalmology, Oulu University Hospital. Thirty-four patients attended the 1-year checkup, and the 2- and 4-year results are available for 31. RESULTS: The pre-operative IOP was 16.0 (SD 4.3, range 6-25) mmHg in the morning and 16.4 (SD 4.0, range 8-25.5) mmHg in the afternoon. On the first postoperative day, the IOP was 2.1 +/- 5.6 mmHg higher than before surgery (p = 0.029). At 1 month, the IOP morning measurement had decreased 2.8 +/- 3.6 mmHg, and in the afternoon, the decrease was 3.0 +/- 2.7 mmHg from the pre-operative level. At 6 months, the decrease was 3.3 +/- 2.7 mmHg in the morning and 3.6 +/- 2.7 mmHg in the afternoon, at 1 year, 3.2 +/- 3.0 mmHg and 3.5 +/- 3.2 mmHg, at 2 years, 3.2 +/- 2.4 mmHg and 3.1 +/- 2.8 mmHg, and at the 4-year postoperative checkup, 3.6 +/- 3.4 mmHg and 3.6 +/- 2.7 mmHg, respectively (p = 0.000 for all time-points). CONCLUSIONS: IOP decreases by about 3 mmHg (16-23% from the pre-operative IOP level) after phacoemulsification and remains at this reduced level with no trend towards an increase during 4 years. PMID- 20003108 TI - Comparison of retinal thickness and fundus-related microperimetry with visual acuity in uveitic macular oedema. AB - PURPOSE: Macular oedema is a common complication and vision-limiting factor in uveitis. The aim of this study was to compare retinal thickness as measured by optical coherence tomography and photoreceptor function as measured by fundus related microperimetry with respect to their correlation with visual acuity. METHODS: Prospective observational monocentre study. Thirty-one patients (53 eyes) with endogenous uveitis and fluorescein angiographically confirmed macular oedema were evaluated. Foveal thickness was analysed using spectral-domain (Spectralis(TM) ; Heidelberg Engineering, Heidelberg, Germany) OCT and retinal sensitivity was assessed using fundus-related microperimetry (MP1; Nidek Technologies, Padova, Italy). All findings were correlated with best-corrected visual acuity (BCVA). RESULTS: Foveal thickness was correlated with BCVA [p = 0.005, r = 0.38, 95% confidence interval (CI) 0.12-0.59]. For microperimetry measurements, a negative correlation with logMAR visual acuity was found. Fixation abnormalities were not associated with poor visual acuity, increased foveal thickness or retinal sensitivity. In eyes with cystoid changes in the outer plexiform and inner nuclear layer, foveal thickness was increased (p < 0.0001). Epiretinal membrane formation was present in 70%. In these eyes, foveal thickness was significantly increased (p = 0.003) and visual acuity was worse (p = 0.08). CONCLUSION: Foveal thickness and fundus-related microperimetry were correlated with visual acuity. Cystoid changes in the outer plexiform and inner nuclear layer and the presence of epiretinal membrane were associated with poor visual acuity. Fixation abnormalities were not associated with poor visual acuity. PMID- 20003110 TI - Terson's syndrome as a prognostic factor for mortality of spontaneous subarachnoid haemorrhage. AB - PURPOSE: To evaluate the prognosis of mortality in patients with spontaneous subarachnoid haemorrhage associated with Terson's syndrome. METHODS: A prospective, consecutive case series study was conducted in patients admitted to the emergency room with a diagnosis of acute subarachnoid haemorrhage. After a complete neurological examination, funduscopic examination using binocular indirect ophthalmoscopy under mydriasis was performed upon admission and at days 3, 7, 30 and 60 after the onset. In all cases, the diagnosis of intracranial bleeding was made by computerized tomography, and the clinical condition was graded according to the Hunt & Hess and Glasgow coma scales. RESULTS: Forty-seven patients with the diagnosis of subarachnoid haemorrhage were enrolled. Forty-four cases were associated with a ruptured aneurysm and three cases with arterio venous malformation. Fourteen patients (29%) were diagnosed with Terson's syndrome. Seven patients (50%) with Terson's syndrome died, whereas death occurred in three patients (9%) without Terson's syndrome (p = 0.002). Ocular findings in Terson's syndrome were preretinal, intraretinal, sub-retinal and vitreous haemorrhage. Associated ocular findings included third-nerve palsy, papilloedema and subconjunctival haemorrhage. CONCLUSION: The presence of Terson's syndrome was associated with an increased mortality rate (50% versus 9%; p < 0.01). Therefore, patients with the diagnosis of intracranial haemorrhage should be submitted to a funduscopic examination, because the presence of intraocular haemorrhage is an important life-threatening prognostic factor. PMID- 20003109 TI - Spontaneous closure of lamellar macular holes studied by optical coherence tomography. AB - PURPOSE: To document and study the spontaneous closure of lamellar macular holes (LMH) by optical coherence tomography (OCT). METHODS: Two women with LMH, 62 and 71 years old, respectively, were followed up with fundoscopy, fundus photography and OCT. RESULTS: In both patients spontaneous closure of LMH was observed 11 and 21 months after baseline examination, respectively. The foveal thickness in case 1 increased from 84 MUm at baseline to 162 MUm at the final examination. The foveal thickness in case 2 increased from 48 MUm at baseline to 148 MUm at the final examination. The foveal contour was also restored in both eyes. The foveal morphology was preserved in both eyes during the follow-up period. CONCLUSION: In both patients the spontaneous closure of LMH could be attributed to the shrinkage of the hole or the release of the tension on the retinal surface, which followed the complete posterior vitreous detachment and separation of epiretinal membrane from the retina. PMID- 20003111 TI - Central retinal vein occlusion following trabeculectomy with OloGen in patients with advanced glaucoma: a possible side-effect? PMID- 20003112 TI - Strongyloides stercoralis hyperinfection: difficulties in diagnosis and treatment. AB - Immunocompromised patients who are infected with Strongyloides stercoralis may develop a potentially fatal auto-infection syndrome characterised by non-specific pulmonary and gastrointestinal symptoms and Gram negative sepsis. We present the case of one such patient who underwent a negative laparotomy for a presumed intra abdominal surgical catastrophe with a subsequent protracted stay on the intensive care unit. Once the diagnosis of strongyloidiasis was made, the patient was successfully treated with subcutaneous antihelminthic drugs. With appropriate screening for and eradication of strongyloides in those with immune compromise, or in those about to start immunosuppressive therapy, potentially fatal episodes of hyperinfection could be avoided. In the absence of screening, severe strongyloidiasis should be suspected in immunosuppressed individuals who have travelled to or resided in an endemic area and present with the characteristic features. Awareness of the signs of hyperinfection amongst those involved in acute care could prevent unnecessary morbidity and mortality in these patients. PMID- 20003113 TI - Elective peri-operative intra-aortic balloon counterpulsation during maxillofacial free flap reconstructive surgery in a patient with severe cardiomyopathy. AB - A key anaesthetic goal in reconstructive free flap surgery is to maintain optimum blood flow to maximise flap survival. We describe the use of intra-aortic balloon counterpulsation in a patient with severe cardiac dysfunction, in whom surgery was initially felt to be contraindicated. PMID- 20003114 TI - Beyond competence: defining and promoting excellence in anaesthesia. AB - Recent trends in medical training have tended to focus on competence, in the sense of adequate performance, rather than excellence. This article reviews published literature and relevant concepts relating to excellence and professionalism from within anaesthesia, from medicine more generally and from outside the profession. A number of conceptual frameworks are presented that could be adapted for the promotion of excellence, and some of the necessary prerequisites for this promotion discussed. PMID- 20003115 TI - Effect of an episode of critical illness on subsequent hospitalisation: a linked data study. AB - Healthcare utilisation can affect quality of life and is important in assessing the cost-effectiveness of medical interventions. A clinical database was linked to two Australian state administrative databases to assess the difference in incidence of healthcare utilisation of 19,921 patients who survived their first episode of critical illness. The number of hospital admissions and days of hospitalisation per patient-year was respectively 150% and 220% greater after than before an episode of critical illness (assessed over the same time period). This was the case regardless of age or type of surgery (i.e. cardiac vs non cardiac). After adjusting for the ageing effect of the cohort as a whole, there was still an unexplained two to four-fold increase in hospital admissions per patient-year after an episode of critical illness. We conclude that an episode of critical illness is a robust predictor of subsequent healthcare utilisation. PMID- 20003116 TI - The cardiovascular effects of normobaric hyperoxia in patients with heart rate fixed by permanent pacemaker. AB - To investigate whether the established reductions in heart rate and cardiac output with hyperoxia in humans are primary effects or secondary to increases in systemic vascular resistance, we paced the hearts of nine patients with permanent pacemakers at a fixed rate when breathing either medical air (inspired O(2) fraction 0.21) or oxygen (inspired O(2) fraction 0.80) in a randomised, double blind fashion. A thoracic bio-impedance machine was used to measure heart rate, stroke volume and blood pressure and calculate cardiac index and systemic vascular resistance index. Oxygen caused no change in cardiac index (p = 0.18), stroke index (p = 0.44) or blood pressure (p = 0.52) but caused a small (5.5%) increase in systemic vascular resistance index (p = 0.03). This suggests that hyperoxia has no direct myocardial depressant effects, but that the changes in cardiac output reported in previous studies are secondary to changes in systemic vascular resistance. PMID- 20003117 TI - The acute hypoxic ventilatory response under halothane, isoflurane, and sevoflurane anaesthesia in rats. AB - The relative order of potency of anaesthetic agents on the hypoxic ventilatory response has been tested in humans, but animal data are sparse. We examined the effects of 1.4, 1.6, 1.8, and 2.0 MAC halothane, isoflurane, and sevoflurane on phrenic nerve activity in euoxia (baseline) and during acute normocapnic hypoxia (inspired oxygen fraction 0.09) in adult male Sprague-Dawley rats. With halothane, all animals became apnoeic even in euoxia, and the hypoxic response was completely abolished at all anaesthetic levels. With isoflurane, 5 of 14 animals exhibited phrenic nerve activity in euoxia at 1.4 MAC and demonstrated a hypoxic response (302% of baseline activity), but all became apnoeic and lost the hypoxic response at higher doses. With sevoflurane, phrenic nerve activity and a hypoxic response was preserved in at least some animals at all doses (i.e. even the highest dose of 2.0 MAC). Similar to the rank order of potency previously observed in humans, the relative order of potency of depression of the hypoxic ventilatory response in rats was halothane (most depressive) > isoflurane > sevoflurane (p = 0.01 for differences between agents). PMID- 20003118 TI - Association of human micro-opioid receptor gene polymorphism A118G with fentanyl analgesia consumption in Chinese gynaecological patients. AB - One hundred and seventy-four Chinese gynaecology patients were studied for the impact of A118G polymorphism in the micro-opioid receptor gene (OPRM1) on pain sensitivity and postoperative fentanyl consumption. Pre-operatively, the pain threshold and pain tolerance threshold were measured using electrical stimulation. A118G polymorphism was genotyped using the polymerase chain reaction restriction fragment length polymorphism method. Intravenous fentanyl patient controlled analgesia provided postoperative pain management, assessed using a visual analogue scale and fentanyl consumed in the first 24 h after surgery was noted. We found the prevalence of G118 allele was 31.3%. The A118G polymorphism had a gene-dose-dependent effect on electrical pain tolerance threshold. Fentanyl consumption was also significantly different in patients with different OPRM1 genotypes (homozygotes for 118G consumed more than did heterozygotes or homozygotes for 118A). Fentanyl consumption increased in accordance with the number of 118G alleles. We conclude that OPRM1 gene analysis may help predict individual opioid sensitivity and so optimise postoperative pain control. PMID- 20003119 TI - Acute hypotension associated with leucocyte depletion filters during cell salvaged blood transfusion. AB - Summary Intra-operative cell salvage is used in more than 75% of NHS hospitals in the United Kingdom and is a safe and cost effective alternative to allogenic blood transfusion. We report a case of acute hypotension during reinfusion of cell salvaged blood through a leucocyte depletion filter that occurred during a caesarean section. We review the literature of hypotension associated with the use of bedside leucocyte depletion filters. PMID- 20003121 TI - Validation of the Wong-Baker FACES Pain Rating Scale in pediatric emergency department patients. AB - OBJECTIVES: The Wong-Baker FACES Pain Rating Scale (WBS), used in children to rate pain severity, has been validated outside the emergency department (ED), mostly for chronic pain. The authors validated the WBS in children presenting to the ED with pain by identifying a corresponding mean value of the visual analog scale (VAS) for each face of the WBS and determined the relationship between the WBS and VAS. The hypothesis was that the pain severity ratings on the WBS would be highly correlated (Spearman's rho > 0.80) with those on a VAS. METHODS: This was a prospective, observational study of children ages 8-17 years with pain presenting to a suburban, academic pediatric ED. Children rated their pain severity on a six-item ordinal faces scale (WBS) from none to worst and a 100-mm VAS from least to most. Analysis of variance (ANOVA) was used to compare mean VAS scores across the six ordinal categories. Spearman's correlation (rho) was used to measure agreement between the continuous and ordinal scales. RESULTS: A total of 120 patients were assessed: the median age was 13 years (interquartile range [IQR] = 10-15 years), 50% were female, 78% were white, and six patients (5%) used a language other than English at home. The most commonly specified locations of pain were extremity (37%), abdomen (19%), and back/neck (11%). The mean VAS increased uniformly across WBS categories in increments of about 17 mm. ANOVA demonstrated significant differences in mean VAS across face groups. Post hoc testing demonstrated that each mean VAS was significantly different from every other mean VAS. Agreement between the WBS and VAS was excellent (rho = 0.90; 95% confidence interval [CI] = 0.86 to 0.93). There was no association between age, sex, or pain location with either pain score. CONCLUSIONS: The VAS was found to have an excellent correlation in older children with acute pain in the ED and had a uniformly increasing relationship with WBS. This finding has implications for research on pain management using the WBS as an assessment tool. PMID- 20003120 TI - Inferior vena cava percentage collapse during respiration is affected by the sampling location: an ultrasound study in healthy volunteers. AB - OBJECTIVES: Physicians are unable to reliably determine intravascular volume status through the clinical examination. Respiratory variation in the diameter of the inferior vena cava (IVC) has been investigated as a noninvasive marker of intravascular volume status; however, there has been a lack of standardization across investigations. The authors evaluated three locations along the IVC to determine if there is clinical equivalence of the respiratory percent collapse at these sites. The objective of this study was to determine the importance of location when measuring the IVC diameter during quiet respiration. METHODS: Measurements of the IVC were obtained during quiet passive respiration in supine healthy volunteers. All images were recorded in B-mode, with cine-loop adjustments in real time, to ensure that maximum and minimum IVC dimensions were obtained. One-way repeated-measures analysis of variance (ANOVA) was used for comparison of IVC measurement sites. RESULTS: The mean (+/-SD) percentage collapse was 20% (+/-16%) at the level of the diaphragm, 30% (+/-21%) at the level of the hepatic vein inlet, and 35% (+/-22%) at the level of the left renal vein. ANOVA revealed a significant overall effect for location of measurement, with F(2,35) = 6.00 and p = 0.006. Contrasts showed that the diaphragm percentage collapse was significantly smaller than the hepatic (F(1,36) = 5.14; p = 0.03) or renal caval index (F(1,36) = 11.85; p = 0.002). CONCLUSIONS: Measurements of respiratory variation in IVC collapse in healthy volunteers are equivalent at the level of the left renal vein and at 2 cm caudal to the hepatic vein inlet. Measurements taken at the junction of the right atrium and IVC are not equivalent to the other sites; clinicians should avoid measuring percentage collapse of the IVC at this location. PMID- 20003122 TI - Assessment of a new model for femoral ultrasound-guided central venous access procedural training: a pilot study. AB - OBJECTIVES: Repetitive practice with feedback in residency training is essential in the development of procedural competency. Lightly embalmed cadaver laboratories provide excellent simulation models for a variety of procedures, but to the best of our knowledge, none describe a central venous access model that includes the key psychomotor feedback elements for the procedure, namely intravascular contents that allow for determination of correct needle position by either ultrasonographic imaging and/or aspiration or vascular contents. METHODS: A cadaver was lightly embalmed using a technique that preserves tissue texture and elasticity. We then performed popliteal fossa dissections exposing the popliteal artery and vein. Vessels were ligated distally, and 14-gauge catheters were introduced into the lumen of each artery and vein. The popliteal artery and vein were then infused with 200 mL of icterine/gel and 200 mL of methylene blue/gel, respectively. Physician evaluators then performed ultrasound (US) guided femoral central venous line placements and rated the key psychomotor elements on a five-point Likert scale. RESULTS: The physician evaluators reported a median of 10.5 years of clinical emergency medicine (EM) experience with an interquartile range (IQR) of 16 and a median of 10 central lines placed annually (IQR = 10). Physician evaluators rated the key psychomotor elements of the simulated procedure as follows: ultrasonographic image of vascular elements, 4 (IQR = 0); needle penetration of skin, 4.5 (IQR = 1); needle penetration of vein, 5 (IQR = 1); US image of needle penetrating vein, 4 (IQR = 2); aspiration of vein contents, 3 (IQR = 2); passage of dilator into vein, 4 (IQR = 2); insertion of central venous catheter, 5 (IQR = 1); US image of catheter insertion into vein, 5 (IQR = 1); and overall psychomotor feedback of the simulated procedure compared to the evaluators' actual patient experience, 4 (IQR = 1). CONCLUSIONS: For the key psychomotor elements of central venous access, the lightly embalmed cadaver with intravascular water-soluble gel infusion provided a procedural model that closely simulated clinicians' experience with patients. PMID- 20003123 TI - Recent caffeine ingestion reduces adenosine efficacy in the treatment of paroxysmal supraventricular tachycardia. AB - OBJECTIVES: Caffeine, an adenosine receptor blocker, should theoretically reduce adenosine efficacy in the treatment of paroxysmal supraventricular tachycardia (SVT). We aimed to determine the effect of recent caffeine ingestion on the likelihood of reversion of SVT with adenosine. METHODS: This was a multicenter, case-control study of adult patients with SVT treated with adenosine between September 2007 and July 2008. The primary endpoint was reversion to sinus rhythm (SR) after a 6-mg adenosine bolus, as a function of recent (within 2, 4, 6, and 8 hours) caffeine ingestion. Caffeine ingestion data were collected using a self administered questionnaire. RESULTS: Of 68 patients enrolled, 52 (76.5%, 95% confidence interval [CI] = 64.4% to 85.6%) reverted after a 6-mg adenosine bolus. There were no significant differences in age, sex, or daily caffeine ingestion between patients who did and did not revert (p > 0.05). However, as a group, patients who did not revert had recently ingested significantly more caffeine (p < 0.05). If caffeine had been ingested less than 2 or 4 hours before the adenosine bolus, the odds of reversion to SR were significantly reduced (odds ratio [OR] = 0.18, 95% CI = 0.04 to 0.93; and OR = 0.14, 95% CI = 0.04 to 0.49, respectively). If caffeine had been ingested less than 6 or 8 hours before the adenosine, the odds of reversion were not reduced (OR = 0.31, 95% CI = 0.09 to 1.02; and OR = 0.31, 95% CI = 0.09 to 1.08, respectively). CONCLUSIONS: Ingestion of caffeine less than 4 hours before a 6-mg adenosine bolus significantly reduces its effectiveness in the treatment of SVT. An increased initial adenosine dose may be indicated for these patients. PMID- 20003124 TI - Predictive ability of propofol effect-site concentrations during fast and slow infusion rates. AB - BACKGROUND: The performance of propofol effect-site pharmacokinetic models during target-controlled infusion (TCI) might be affected by propofol administration rate. This study compares the predictive ability of three effect-site pharmacokinetic models during fast and slow infusion rates, utilizing the cerebral state index (CSI) as a monitor of consciousness. METHODS: Sixteen healthy volunteers, 21-45 years of age, were randomly assigned to receive either a bolus dose of propofol 1.8 mg/kg at a rate of 1200 ml/h or an infusion of 12 mg/kg/h until 3-5 min after loss of consciousness (LOC). After spontaneous recovery of the CSI, the bolus was administered to patients who had first received the infusion and vice versa. The study was completed after spontaneous recovery of CSI following the second dose scheme. LOC was assessed and recorded when it occurred. Adequacies of model predictions during both administration schemes were assessed by comparing the effect-site concentrations estimated at the time of LOC during the bolus dose and during the infusion scheme. RESULTS: LOC occurred 0.97 +/- 0.29 min after the bolus dose and 6.77 +/- 3.82 min after beginning the infusion scheme (P<0.05). The Ce estimated with Schnider (ke0=0.45/min), Marsh (ke0=1.21/min) and Marsh (ke0=0.26/min) at LOC were 4.40 +/ 1.45, 3.55 +/- 0.64 and 1.28 +/- 0.44 microg/ml during the bolus dose and 2.81 +/- 0.61, 2.50 +/- 0.39 and 1.72 +/- 0.41 microg/ml, during the infusion scheme (P<0.05). The CSI values observed at LOC were 70 +/- 4 during the bolus dose and 71 +/- 2 during the infusion scheme (NS). CONCLUSION: Speed of infusion, within the ranges allowed by TCI pumps, significantly affects the accuracy of Ce predictions. The CSI monitor was shown to be a useful tool to predict LOC in both rapid and slow infusion schemes. PMID- 20003126 TI - Regional cerebral blood flow responses to hyperventilation during sevoflurane anaesthesia studied with PET. AB - BACKGROUND: Arterial carbon dioxide tension (PaCO(2)) is an important factor controlling cerebral blood flow (CBF) in neurosurgical patients. It is still unclear whether the hypocapnia-induced decrease in CBF is a general effect on the brain or rather linked to specific brain regions. We evaluated the effects of hyperventilation on regional cerebral blood flow (rCBF) in healthy volunteers during sevoflurane anaesthesia measured with positron emission tomography (PET). METHODS: Eight human volunteers were anaesthetized with sevoflurane 1 MAC, while exposed to hyperventilation. During 1 MAC sevoflurane at normocapnia and 1 MAC sevoflurane at hypocapnia, one H(2)(15)O scan was performed. Statistical parametric maps and conventional regions of interest analysis were used for estimating rCBF differences. RESULTS: Cardiovascular parameters were maintained constant over time. During hyperventilation, the mean PaCO(2) was decreased from 5.5 + or - 0.7 to 3.8 + or - 0.9 kPa. Total CBF decreased during the hypocapnic state by 44%. PET revealed wide variations in CBF between regions. The greatest values of vascular responses during hypocapnia were observed in the thalamus, medial occipitotemporal gyrus, cerebellum, precuneus, putamen and insula regions. The lowest values were observed in the superior parietal lobe, middle and inferior frontal gyrus, middle and inferior temporal gyrus and precentral gyrus. No increases in rCBF were observed. CONCLUSIONS: This study reports highly localized and specific changes in rCBF during hyperventilation in sevoflurane anaesthesia, with the most pronounced decreases in the sub cortical grey matter. Such regional heterogeneity of the cerebral vascular response should be considered in the assessment of cerebral perfusion reserve during hypocapnia. PMID- 20003127 TI - Dexmedetomidine provides cortical neuroprotection: impact on anaesthetic-induced neuroapoptosis in the rat developing brain. AB - BACKGROUND: Recent evidence has demonstrated the anti-apoptotic of dexmedetomidine in different brain injury models. Herein, we investigated whether dexmedetomidine could directly protect against cortical injury in vitro and in vivo. METHODS: Apoptosis was induced by staurosporine or wortmannin treatment in cortical neuronal cultures in vitro or by 6 h of isoflurane (0.75%) administration to post-natal day 7 rat pups in vivo. Dexmedetomidine was then applied in escalating doses to assess the neuroprotective potential of this agent. Cell survival was quantified using an MTT assay in vitro and in vivo apoptosis was assessed using cleaved caspase-3 immunohistochemistry. Cortical Western blots were conducted for the cellular survival proteins Bcl-2 and phosphorylated extracellular signal-regulated protein kinase (pERK)1 and 2. RESULTS: In vitro dexmedetomidine dose-dependently prevented both staurosporine- and wortmannin-induced injury in cortical neuronal cultures, indicating that dexmedetomidine can prevent apoptosis when applied directly. In vivo isoflurane induced cortical neuroapoptosis compared with air (327+/-80 vs. 34+/-9 caspase-3 positive neurons; P<0.05). Dexmedetomidine inhibited isoflurane-induced caspase-3 expression (P<0.05), although the protection achieved did not completely attenuate the isoflurane injury (P<0.05 vs. air). Isoflurane treatment decreased Bcl-2 and pERK protein expression relative to air, an effect reversed by dexmedetomidine treatment. CONCLUSIONS: Dexmedetomidine prevents cortical apoptosis in vitro and in vivo. However, using higher doses of dexmedetomidine does not further increase protection against isoflurane injury in the cortex than previously observed. PMID- 20003128 TI - Distribution of ABO blood group allele and identification of three novel alleles in the Chinese Han population. AB - BACKGROUND: The ABO blood group system is clinically important in blood transfusion. The molecular characterization of ABO blood group has clinical and anthropological importance. Here, we determined the ABO alleles distribution and identified three novel alleles in the Chinese Han population. STUDY DESIGN AND METHODS: Four hundred and seventeen Chinese Han individuals were determined by standard serologic techniques for the ABO blood group phenotypes. The ABO genotypes and alleles were analysed by polymerase chain reaction sequence-based typing (PCR-SBT) for sequencing exon 6 to 7 of the ABO gene. The polymorphisms of intron 5 and 6 of the ABO gene were also analysed by PCR-SBT. The two haplotypes including new alleles were separated by a Dynabeads M-270 Streptavidin protocol. RESULTS: All ABO genotypes of the samples were consistent with the phenotypes. Fourteen alleles were identified based on the nucleotide sequences of exon 6 and 7, with five common alleles (A101, A102, B101, O01 and O02), six known rare alleles (A205, B110, O04, O05, O07 and O50) and three novel alleles (B112, CisAB06 and 061). The three new alleles appeared with the frequencies of 0.12%, 0.12% and 0.36%, respectively. CONCLUSION: The detailed frequencies distribution of ABO alleles was studied in the Chinese Han population. We identified 14 alleles, including 3 novel alleles. PMID- 20003130 TI - Collaborative study to establish a replacement World Health Organization International Standard for parvovirus B19 DNA nucleic acid amplification technology (NAT)-based assays. AB - BACKGROUND AND OBJECTIVES: The aim of the study was to replace the 1(st) World Health Organization International Standard for parvovirus B19 DNA for nucleic acid amplification technique (NAT)-based assays (code 99/800). Two lyophilized preparations (coded 99/800 and 99/802) had been evaluated in the original collaborative study. The present study re-evaluates these two preparations in terms of potency, stability and encapsidation of virus DNA. MATERIALS AND METHODS: The 1(st) International Standard (99/800) and 99/802 were re-coded as Samples 1 and 2, respectively. The samples were distributed to six laboratories and assayed on four separate occasions. Accelerated thermal degradation samples of the two preparations were examined after storage at 20 degrees C for 7 years. Nuclease treatment was used to investigate the encapsidation of virus DNA. RESULTS: Data were returned from a total of six different quantitative NAT-based assays. The results of the present study confirm those of the original, with no significant differences found in estimated international units (IU)/ml for the 1(st) International Standard (Sample 1 in this study) and the proposed replacement preparation, Sample 2 (99/802). Accelerated thermal degradation studies demonstrate that both samples are very stable, with no loss of potency after storage at 20 degrees C for 7 years. Both lyophilized preparations contained the majority of B19V DNA encapsidated in virions. CONCLUSIONS: On the basis of the data presented in this collaborative study, Sample 2 (code number 99/802) was established as the 2(nd) International Standard for parvovirus B19 DNA for NAT-based assays with a potency of 10(6) IU/ml (500 000 IU/vial). PMID- 20003131 TI - Approach to blood donors with microcytosis. AB - The aim of this paper is to find microcytosis in donors, to establish prevalence of iron deficiency anaemia (IDA) and beta-thalassemia trait (BTT) in them and to evaluate which index is most effective in differentiating these two conditions. IDA and BTT are the most common causes of microcytic anaemia. Traditional approach is trial of iron treatment. Where thalassemias are common, this can lead to iron overload and failure to provide diagnosis/counselling in BTT. Initially 925 donor samples were evaluated on cell counter. Of these, 50 were found microcytic. These were subjected to Ferritin and HbA2 determination. Subsequently, additional 51, age- and sex-matched normocytic donor samples were selected as controls. These were subjected to the same tests. Nine indices namely RBC, RDW, Mentzer's, Shine and Lal, England and Fraser, Srivastava, Green and King, RDW index and Ricerca were used to differentiate IDA and BTT. Prevalence of microcytosis was 5.4%. Of these microcytic samples, 52% were IDA, 36% were BTT, 8% had both and 4% were undiagnosed. IDA had significantly lower Hb, mean corpuscular volume (MCV) and Ferritin levels than the control group. BTT had lower MCV, higher Ferritin and comparable Hb levels with control group. The Youden's index of Mentzer's was highest and RBC was the only index which had both sensitivity and specificity more than 80% for both IDA and BTT. It is desirable to routinely perform hemograms for all blood donors and further analyse the microcytic samples for Ferritin and HbA2 to diagnose IDA and BTT and to provide appropriate counselling/treatment. PMID- 20003132 TI - Targeted forward mutagenesis by transitive RNAi. AB - A novel technique is described that targets specific populations of transcripts for homology-based gene silencing using transitive RNAi. This approach is designed to target a subset of the transcriptome in order to identify genes involved in a particular localized process, such as photosynthesis. As a proof-of concept approach, mesophyll cells from Arabidopsis thaliana were laser microdissected from whole leaves to generate a focused cDNA library that was bi directionally cloned into a transitive RNAi vector that had been designed to induce silencing of homologous, endogenous genes. Approximately 15% of the transformant plants identified from both sense and antisense libraries exhibited visible phenotypes indicative of photosynthetic defects. Amplification from the genome and sequencing of cDNA inserts identified candidate genes underlying the phenotypes. For 10 of 11 such mutants, re-transformation with an RNAi construct corresponding to the candidate gene recapitulated the original mutant phenotype, and reduction of corresponding endogene transcripts was confirmed. In addition, one of the re-transformed transgenes also silenced transcripts of closely related family members, thereby demonstrating the utility of this approach for mutagenesis of redundant gene functions. Preliminary results using tissue specific transitive RNAi forward mutagenesis of the Arabidopsis vegetative shoot apical meristem demonstrate the broad applicability of this forward mutagenesis technique for a variety of plant cell types. PMID- 20003133 TI - Two functionally distinct members of the MATE (multi-drug and toxic compound extrusion) family of transporters potentially underlie two major aluminum tolerance QTLs in maize. AB - Crop yields are significantly reduced by aluminum (Al) toxicity on acidic soils, which comprise up to 50% of the world's arable land. Al-activated release of ligands (such as organic acids) from the roots is a major Al tolerance mechanism in plants. In maize, Al-activated root citrate exudation plays an important role in tolerance. However, maize Al tolerance is a complex trait involving multiple genes and physiological mechanisms. Recently, transporters from the MATE family have been shown to mediate Al-activated citrate exudation in a number of plant species. Here we describe the cloning and characterization of two MATE family members in maize, ZmMATE1 and ZmMATE2, which co-localize to major Al tolerance QTL. Both genes encode plasma membrane proteins that mediate significant anion efflux when expressed in Xenopus oocytes. ZmMATE1 expression is mostly concentrated in root tissues, is up-regulated by Al and is significantly higher in Al-tolerant maize genotypes. In contrast, ZmMATE2 expression is not specifically localized to any particular tissue and does not respond to Al. [(14)C]-citrate efflux experiments in oocytes demonstrate that ZmMATE1 is a citrate transporter. In addition, ZmMATE1 expression confers a significant increase in Al tolerance in transgenic Arabidopsis. Our data suggests that ZmMATE1 is a functional homolog of the Al tolerance genes recently characterized in sorghum, barley and Arabidopsis, and is likely to underlie the largest maize Al tolerance QTL found on chromosome 6. However, ZmMATE2 most likely does not encode a citrate transporter, and could be involved in a novel Al tolerance mechanism. PMID- 20003134 TI - Probing plant membranes with FM dyes: tracking, dragging or blocking? AB - Remarkable progress in various techniques of in vivo fluorescence microscopy has brought an urgent need for reliable markers for tracking cellular structures and processes. The goal of this manuscript is to describe unexplored effects of the FM (Fei Mao) styryl dyes, which are widely used probes that label processes of endocytosis and vesicle trafficking in eukaryotic cells. Although there are few reports on the effect of styryl dyes on membrane fluidity and the activity of mammalian receptors, FM dyes have been considered as reliable tools for tracking of plant endocytosis. Using plasma membrane-localized transporters for the plant hormone auxin in tobacco BY-2 and Arabidopsis thaliana cell suspensions, we show that routinely used concentrations of FM 4-64 and FM 5-95 trigger transient re localization of these proteins, and FM 1-43 affects their activity. The active process of re-localization is blocked neither by inhibitors of endocytosis nor by cytoskeletal drugs. It does not occur in A. thaliana roots and depends on the degree of hydrophobicity (lipophilicity) of a particular FM dye. Our results emphasize the need for circumspection during in vivo studies of membrane proteins performed using simultaneous labelling with FM dyes. PMID- 20003135 TI - Molecular analysis of HLA Class I and Class II genes in four indigenous Malaysian populations. AB - This is the first report of high-resolution human leukocyte antigen (HLA) typing in four indigenous groups in Malaysia. A total of 99 normal, healthy participants representing the Negrito (Jehai and Kensiu), Proto-Malay (Temuan) and a native group of Borneo (Bidayuh) were typed for HLA-A, -B, -DRB1 and -DQB1 genes using sequence-based typing. Eleven HLA-A, 26 HLA-B, 16 HLA-DRB1 and 14 HLA-DQB1 alleles were detected, including a new allele, HLA-B*3589 in the Jehai. Highly frequent alleles were A*2407, B*1513, B*1801, DRB1*0901, DRB1*1202, DRB1*1502, DQB1*0303 and DQB1*0502. Principal component analysis based on high-resolution HLA-A, -B and -DRB1 allele frequencies showed close affinities among all four groups, including the Negritos, with other Southeast Asian populations. These results showed the scope of HLA diversity in these indigenous minority groups and may prove beneficial for future disease association, anthropological and forensic studies. PMID- 20003136 TI - Fine epitope mapping of monoclonal antibodies 9B9 and 3G8 to the N domain of angiotensin-converting enzyme (CD143) defines a region involved in regulating angiotensin-converting enzyme dimerization and shedding. AB - A panel of monoclonal antibodies (mAbs) raised against both the N and C domains of angiotensin-I-converting enzyme (ACE, peptidyl dipeptidase, EC 3.4.15.2) have been extensively mapped and have facilitated the study of various aspects of ACE structure and biology. In this study, we characterize two mAbs, 9B9 and 3G8, that recognize the N domain of ACE and that influence shedding and dimerization. Fine epitope mapping was performed, which mapped the epitopes for these mAbs to the N terminal region of the N domain where they overlap to a large extent, despite having different effects on ACE processing. The mAb 3G8 epitope appears to be shielded by the C domain and to be carbohydrate dependent as binding increased significantly as a result of underglycosylation, whereas these factors did not influence mAb 9B9 recognition. Three mutations within the overlapping region of these two epitopes, Q18H, L19E, and Q22A, which decreased mAb 3G8 binding to the soluble N domain, were introduced into full-length somatic ACE (sACE) to determine their influence on ACE expression and processing. Increased ACE expression, cell surface expression, and basal shedding were observed with all three mutations. Furthermore, cross-linking and western blotting of Chinese hamster ovary (CHO) cell lysates detected two distinct ACE dimers, a native and cross-linked dimer. Increasing amounts of the cross-linked dimer were observed for the mutant sACEQ22A, further implicating the overlapping region of the mAb 9B9 and 3G8 epitopes in ACE processing. PMID- 20003137 TI - HLA alleles and haplotypes distribution in Dai population in Yunnan province, Southwest China. AB - Human leukocyte antigen (HLA) analysis would be a useful tool to trace the origin of modern humans. In this study, we provided the first four digital HLA-A, -B, -C and -DRB1 allele and haplotype data in the Dai ethnic population, which is a unique and representative Kam-Tai-speaking ethnic minority living in the Yunnan province of Southwestern China. Our results showed that the Dai population has unique HLA characteristic that are most closely related to the Southeastern Asia group and similar to the Kam-Tai speaking populations in China and Thailand. PMID- 20003138 TI - Biliary strictures in pediatric liver transplant recipients - early diagnosis and treatment results in excellent graft outcomes. AB - Biliary complications in pediatric LT are important causes of morbidity and graft loss. We examined our recent pediatric LT experience to determine the outcome of post-LT biliary complications and their relationship to graft type. All initially isolated LTs performed at our institution between January 1, 2000 and August 20, 2007 were reviewed. Recipient data, donor type, graft survival, and biliary complications data were examined. Of 66 LTs, 32 patients received whole organ grafts, and 34 received partial grafts; 11 split, seven reduced size, and 16 live donors. Seventy-seven percent of patients had biliary reconstruction using a RYH. Overall, 17 (26%) developed biliary complications, and 15 were diagnosed within six months post-LT. Live donor and split allografts had more biliary complications than reduced size or whole allografts (50% and 36% vs. 0% and 16%, respectively). Seventy-one percent responded to percutaneous or endoscopic treatment. Five failed initial non-operative management and required reoperation (one retransplantation). These data suggest that biliary strictures occur most frequently in live donor and split allografts and that non-operative therapy is highly successful. Partial grafts are essential in pediatric LT, and a high clinical suspicion for biliary complications combined with aggressive and early diagnosis and therapy rarely results in graft loss. PMID- 20003139 TI - Anal-transitional and bronchial metaplasia within the ileostomy of a small bowel allograft: a case report. AB - Epithelial metaplasia is a phenomenon where a native mucosal cell type is replaced by another due to altered stem cell differentiation, usually as the result of long-standing injury or inflammation. It is common in ileostomies performed with native small bowels for both neoplastic and inflammatory conditions. We present a case of anal-transitional and bronchial epithelial metaplasia that occurred near the ileostomy of a small bowel allograft in a seven month-old male transplanted for short bowel syndrome related to gastroschisis. The anal-transitional metaplastic mucosa had an immunophenotype (CK7- and p63 positive) similar to that of normal mucosa at the anal transition zone and a DNA STR profile consistent with the donor. To our knowledge, this represents the first report of this type of metaplasia occurring in ileostomies in the English literature. It is important for clinicians and pathologists to recognize these benign mucosal alterations to avoid diagnostic confusion. PMID- 20003140 TI - Thyroid function abnormalities among first-degree relatives of Iranian congenital hypothyroidism neonates. AB - BACKGROUND: Congenital hypothyroidism (CH) is a relatively common metabolic disease in neonates. Until recent years the disorder was usually regarded as occurring in a sporadic manner. Over the past few years, however, a considerable proportion of familial cases have been identified, and possible roles of autoimmune factors suggested. The aim of the present study was to evaluate abnormality of thyroid function tests in first-degree relatives of CH neonates and compared this to the normal population. METHODS: From 2002 until 2007 thyroid function tests (T4 and thyroid-stimulating hormone [TSH]) were done in randomly selected CH and normal neonates (n= 194 and n= 350, respectively) and their first degree relatives. Most mothers of the CH neonates and control groups were also evaluated for thyroid peroxidase antibody (TPOAb). RESULTS: Thyroid function test in first-degree relative of neonates with CH (361 parents, 136 siblings) were compared with those in control groups (665 parents, 478 siblings). Abnormal thyroid function tests were found in 85 patients in the CH group versus 96 patients in the control group; hypothyroidism was found in 75 (15.1%) and 57 subjects (5%) person in the CH and control groups, respectively (P < 0.05). Positive TPO antibody was found in 22 mothers (17.3%) of CH neonates in comparison with 65 mothers (32.5%) of control groups (P < 0.05). Frequency of hyperthyroidism in parents of control group had trend to be higher than parents of CH neonates (P= 0.05) CONCLUSION: Familial and genetic components play a role in inheritance of CH, but maternal thyroid autoimmunity may not play an important role in the development of CH in Iran. PMID- 20003141 TI - Colistimethate sodium therapy for multidrug-resistant isolates in pediatric patients. AB - AIM: The aim of the present study was to assess the efficacy and safety of colistimethate sodium therapy in multidrug-resistant nosocomial infections caused by Pseudomonas aeruginosa or Acinetobacter baumannii in neonates and children. METHODS: Pediatric patients hospitalized at the Uludag University Hospital who had nosocomial infections caused by multidrug-resistant P. aeruginosa or A. baumannii, were enrolled in the study. Colistimethate sodium at a dosage of 50-75 x 10(3) U/kg per day was given i.v. divided into three doses. RESULTS: Fifteen patients received 17 courses of colistimethate sodium for the following infections: ventilator-associated pneumonia (n= 14), catheter-related sepsis (n= 1) and skin and soft-tissue infection (n= 2). The mean age of patients was 53.2 + 74.7 months (range, 8 days-15 years) and 60% were male. Mortality was 26.6%. CONCLUSION: Colistimethate sodium appears to be safe and effective for the treatment of severe infections caused by multidrug-resistant P. aeruginosa or A. baumannii in pediatric patients. PMID- 20003142 TI - Implication of early-onset biliary atresia and extrahepatic congenital anomalies. AB - BACKGROUND: The aim of the present study was to determine the rate of early-onset biliary atresia (BA) and its implications, for embryonic-type BA in Taiwan, a high-prevalence area for BA. The relationship between the timing of disease onset and congenital extrahepatic anomalies was also identified. METHODS: Medical records of 130 infants born in Taiwan with biliary atresia between January 1996 and December 2005 were reviewed retrospectively. The gold standard for the diagnosis of biliary atresia was intraoperative cholangiography. As well as medical records review, abdominal imaging and echocardiograms were performed to determine other structural anomalies. Early-onset BA was defined as acholic stool and cholestatic jaundice observed before 2 weeks of age. RESULTS: On review of onset of acholic stool and cholestatic jaundice before 2 weeks of age, 31 patients (23.8%) were defined as having early-onset BA. Twenty patients (15.4%) had major congenital extrahepatic anomalies. One (0.7%) had biliary atresia splenic malformation syndrome (BASM). Both early-onset and late-onset BA may be associated with other structural anomalies. Patients with early-onset BA had a higher probability of having major extrahepatic anomaly (9/31 vs 11/99, P = 0.046). Situs anomalies accompanying major gastrointestinal (GI) tract anomalies occurred only in early-onset BA patients. CONCLUSIONS: After comprehensively investigating the timing of onset and associated congenital extrahepatic anomalies in BA patients in Taiwan, only one BASM with double spleen was detected. A total of 23.8% of patients had early-onset BA, and this group of patients is prone to extrahepatic anomalies. Situs anomalies accompanying major GI tract anomaly may be indicative of embryonic-type early-onset BA. PMID- 20003143 TI - Blood concentrations of midazolam in status epilepticus using an appropriate condition of HPLC. AB - BACKGROUND: The aim of the present study was to determine an index to evaluate the efficacy and safety of midazolam (MDZ) to treat status epilepticus (SE). An original system was therefore developed to measure blood concentrations of MDZ and 1-hydroxymidazolam (1-OHMDZ) as the main metabolite on high-performance liquid chromatography. METHODS: This system was established through inspection of chromatograms, calibration curves and coefficient of correlations of MDZ. The clinical course of 11 SE patients, ranging from 4 months to 10 years of age, are described. These patients were treated with MDZ and measured at each blood concentration of MDZ. Moreover, patients were evaluated on cranial computed tomography and magnetic resonance imaging and video electroencephalogram (EEG), and it was determined that their seizures disappeared in accordance with the disappearance of convulsions and interictal EEG findings. RESULTS: Reproducibility was good with this system. The standard curves of MDZ and 1-OHMDZ were almost straight, and the correlation coefficients of MDZ and 1-OHMDZ were r = 0.9999 and r = 0.9998, respectively. The convulsions in nine of 11 SE patients disappeared without side-effects and the blood concentrations of MDZ in all the patients were measured. The mean peak blood concentrations of MDZ and 1-OHMDZ were higher than those reported in other studies. CONCLUSIONS: The clinical utility of this system has been demonstrated. An index to evaluate the efficacy and safety of MDZ is necessary, and MDZ blood concentrations measured on the present original precise measuring system could help in establishing a plan to successfully treat SE. PMID- 20003144 TI - Attitude to extended use and long-term storage of newborn screening blood spots in Japan. AB - BACKGROUND: Residual dried blood spots (DBS) remaining after routine newborn screening (NBS) tests are candidate specimens for extended uses such as quality assurance and the development of new technology. A trial of NBS using tandem mass spectrometry was launched in 2004 in Japan. The aim of the present study was to analyze the attitudes of the public, patient families, and medical professionals toward the extended use and long-term storage of residual DBS, and to construct a standardized informational brochure. METHODS: A questionnaire was sent to randomly selected members of the public, members of the Japanese Phenylketonuria (PKU) Association, medical staff of a general hospital, staff of a children's hospital, obstetricians and gynecologists, pediatricians and NBS personnel. Associated responses, which were given in a free comment format, were analyzed by text mining. RESULTS: The awareness ratio of NBS was low in the public (26.6%), but despite this, when a brief explanatory note on NBS was provided, 71.7% of them recognized the necessity of NBS. They were less positive than medical professionals and PKU patient families regarding the extended use of DBS for forensic investigation, for the study of health problems, or long-term storage of residual DBS, regardless of whether these factors affected them personally or not. Among the medical professionals, obstetricians and pediatricians exhibited a higher ratio of negative responses toward the extended use and long-term storage of DBS than others. CONCLUSION: The general public is more conservative than PKU patients and their families or medical professionals about the extended use or long-term storage of residual DBS. Presentation to the public, particularly to couples of childbearing age, of appropriate explanatory information on NBS itself, or the extended use or long-term storage of residual DBS, is recommended. PMID- 20003145 TI - Effect of fluid intake on the outcome of constipation in children: PEG 4000 versus lactulose. AB - BACKGROUND: The aim of the present study was to evaluate the effect of fluid intake on the outcome of treatment with osmotic laxatives, PEG 4000 and lactulose, in children with chronic constipation. METHODS: Medical records were reviewed, and 27 children with chronic constipation satisfying the following criteria were enrolled: (i) on stable maintenance medication for >= 3 months; (ii) no dosage modification or drug change between the two periods compared; (iii) no clinical events during the investigation that could affect intake or the clinical outcomes; (iv) good compliance regarding inclusion in the diaries of the frequency of bowel movements, the consistency of stools, and the amount of fluid intake. The diary covered > 25 days per visit; and (v) interval of < 3 months between the two periods compared. A scoring system was developed for bowel movements and for stool consistency. The score during a period of better fluid intake was compared to that during a period of worse fluid intake, for the same patient. Wilcoxon signed-rank test was used for the analysis. RESULTS: On polyethylene glycol 4000 (PEG 4000; n = 14), the bowel movement score on good fluid intake was 21.43-30.00 (median, 27.78), while that on poor fluid intake was 19.00-30.00 (median, 25.13; P = 0.009). For stool consistency, the score on good fluid intake was 11.79-20.00 (median, 20.00), while that on poor fluid intake was 11.00-25.00 (median, 15.91; P = 0.002). On lactulose (n = 13) there was no statistical difference with respect to the two categories of defecation. CONCLUSIONS: Children who are already on PEG 4000 still benefit from additional fluid intake. PMID- 20003146 TI - Effects of discrepancy between imagined and perceived sounds on the N2 component of the event-related potential. AB - Two experiments were conducted to examine whether the N2 component of the event related potential (ERP), typically elicited in a S1-S2 matching task and considered to reflect mismatch process, can still be elicited when the S1 was imagined instead of perceived and to investigate how N2 amplitude varied with the degree of S1-S2 discrepancy. Three levels of discrepancy were defined by the degree of separation between the heard (S2) and imagined (S1) sounds. It was found that the N2 was reliably elicited when the perceived S2 differed from the imagined S1, but whether N2 amplitude increased with the degree of discrepancy depended in part on the S1-S2 discriminability (as evidenced by reaction time). Specifically, the effect of increasing discrepancy was attenuated as discriminability increased from hard to easy. These results, together with the dynamic ERP topography observed within the N2 window, suggest that the N2 effect reflects two sequential but overlapping processes: automatic mismatch and controlled detection. PMID- 20003147 TI - Simplifying jackknifing of ERPs and getting more out of it: retrieving estimates of participants' latencies. AB - Abstract Research has demonstrated that the jackknifing procedure for estimating ERP latencies (J. Miller, T. Patterson, & R. Ulrich, 1998) yields more accurate estimates of differences between experimental conditions in ERP latency than other methods. However, the scores resulting from this procedure require special adjustments for further analyses and do not directly reflect each participant's latency. Here, a simple transform is proposed that retrieves estimates of each participant's latency from the subaverage scores, rendering further adjustments superfluous. Other advantages of working with participants' latencies are discussed. Results of simulations support the validity of jackknifing and the retrieval transform. PMID- 20003148 TI - Task relevance and recognition of concealed information have different influences on electrodermal activity and event-related brain potentials. AB - This study aimed at differentiating between memory- and task-related processes and their correlates on the electrodermal and electrocortical level during information concealment. Variations of the Guilty Knowledge Test were implemented in two experiments while we measured skin conductance responses (SCRs) and event related brain potentials. P300 amplitudes were specifically enhanced for items requiring a deviant behavioral response but they were not sensitive to concealed knowledge. In contrast, N200 amplitudes differed between memorized and irrelevant items in both experiments. SCR measures reflected a combined influence of task relevance and probe recognition, and they provided incremental validity above N200 amplitudes. These results suggest that the P300 mainly reflects task relevance in the given experimental setting whereas the N200 amplitude is sensitive to previously encoded information and potentially linked to response monitoring processes. PMID- 20003149 TI - Sequence effects in cued task switching modulate response preparedness and repetition priming processes. AB - In task-switching paradigms, reaction time (RT) switch cost is eliminated on trials after a no-go trial (no-go/go sequence effect). We examined the locus of no-go interference on task-switching performance by comparing the event-related potential (ERP) time course of go/go and no-go/go sequences from cue onset to response execution. We also examined whether noninformative trials (i.e., delayed reconfiguration, no response inhibition) produce similar sequence effects. Participants switched using informative and noninformative cues (Experiment 2) intermixed with no-go trials (Experiment 1). Repeat RT was slower for both no go/informative (pNG/I) and noninformative/informative (pNI/I) than informative/informative sequences. ERPs linked to anticipatory preparation showed no effect of trial sequence. ERPs indicated that pNG/I sequences reduce response readiness whereas pNI/I sequences reduce repetition benefit for repeat trials. Implications for task-switching models are discussed. PMID- 20003150 TI - Cardiac awareness and autonomic cardiac reactivity during emotional picture viewing and mental stress. AB - This study investigated the relationship between autonomic cardiovascular reactivity and cardiac awareness during the following conditions: baseline, emotional picture viewing, mental stress, and heartbeat tracking. Cardiac parameters were examined by using power spectrum analysis of heart rate variability and impedance cardiography. According to their performance in a heartbeat tracking task, 38 participants were classified as good (n=19) or poor (n=19) heartbeat perceivers. Neither group differed during baseline and heartbeat tracking, but good compared to poor heartbeat perceivers demonstrated greater sympathetic reactivity during mental stress and more vagal reactivity and subjective arousal during emotional picture viewing. The results suggest that cardiac awareness is related to greater responsivity of the autonomic nervous system during situations evoking autonomic reactivity. PMID- 20003151 TI - Direct antifungal effect of femtosecond laser on Trichophyton rubrum onychomycosis. AB - Onychomycosis is caused by dermatophyte infection of the nail. Though laser energy has been shown to eliminate dermatophytes in vitro, direct laser elimination of onychomycosis is not successful due to difficulties in selectively delivering laser energy to the deeper levels of the nail plate without collateral damage. Femtosecond (fsec) infrared titanium sapphire lasers circumvent this problem by the nonlinear interactions of these lasers with biological media. This quality, combined with the deeply penetrating nature of the near-infrared radiation, allows elimination of deeply seeded nail dermatopytes without associated collateral damage. Nail cuttings obtained from patients with onychomycosis caused by Trichophyton rubrum underwent fsec laser irradiation using increasing laser intensities with the focus scanned throughout the whole thickness of the nail specimen. The efficacy of the laser treatment was evaluated by subculture. Scanning electron microscopy was used to determine fsec laser induced collateral damage. We found that a fsec laser fluence of 7 x 10(31) photons m(-2) s(-1) or above successfully inhibited the growth of the fungus in all samples examined, whereas laser intensities above 1.7 x 10(32) photons m(-2) s(-1) affected the structure of the nail plate. Our findings suggest that T. rubrum-mediated onychomycosis may be treated by fsec laser technology. PMID- 20003152 TI - Depth-dependent effects of ultraviolet radiation on survivorship, oxidative stress and DNA damage in sea urchin (Strongylocentrotus droebachiensis) embryos from the Gulf of Maine. AB - A field experiment was conducted on the early embryos of the green sea urchin Strongylocentrotus droebachiensis at different depths in the Gulf of Maine (GOM) to assess the effects of UV radiation (UVR: 300-400 nm) on survivorship, oxidative stress and DNA damage. Embryos experimentally placed at 1 m were exposed to UVB (300-320 nm) where a significant decrease in survivorship was observed as well as significant increases in the activity of the antioxidant enzyme superoxide dismutase and DNA damage. DNA damage includes both cyclobutane pyrimidine dimer photoproducts from direct exposure to UVA (320-400 nm) and indirect DNA damage associated with the production of reactive oxygen species. All embryos had equivalent concentrations of the UVR-absorbing compounds known as mycosporine-like amino acids and despite the fact that these compounds absorb primarily in the UVA portion of the spectrum they did not provide protection for embryos from DNA damage in the field at depths less than 5 m. DNA damage and survivorship of green sea urchin embryos in the GOM was directly related to the optical properties of the water column and the differential attenuation of UVB and UVA wavelengths. PMID- 20003153 TI - Mycosporine-like amino acids in freshwater copepods: potential sources and some factors that affect their bioaccumulation. AB - Mycosporine-like amino acids (MAAs) are ubiquitous photoprotective compounds in aquatic environments. MAAs are synthesized by a wide variety of organisms (i.e. bacteria, fungi and algae) and their production is photoinducible by ultraviolet radiation (UVR) (280-400 nm) and/or photosynthetically active radiation (400-750 nm). Most animals however, are unable to synthesize MAAs and must acquire these compounds through their diet or from symbiotic organisms. In this paper, we investigate the possible sources of MAAs and factors (temperature and initial MAA concentration) that may affect their bioaccumulation in freshwater copepods. We found that MAA accumulation may occur even if the copepods are cultured on a MAA free diet. In addition, we found that the bacteriostatic antibiotic, chloramphenicol, inhibits the bioaccumulation of MAAs. These two pieces of evidence suggest that the source of MAAs in these copepods may be prokaryotic organisms in close association with the animals. The two factors investigated in this study, temperature and initial MAA concentrations, were found to affect the rates at which MAAs are accumulated. Temperature had positive effects on both uptake and elimination rates. On the other hand, the rate of uptake decreased at the highest assayed initial MAA concentration, probably because the concentration of MAAs was already close to saturation. PMID- 20003154 TI - Photocatalysis of chloroform decomposition by hexachloroosmate(IV). AB - Hexachloroosmate(IV) effectively catalyzes the photodecomposition of chloroform in aerated solutions. The decomposition products are consistent with a mechanism in which excited state OsCl(6)(2-) reduces chloroform, rather than one involving photodissociation of chlorine atoms. Trace amounts of ethanol or water in the chloroform lead to photosubstitution to form OsCl(5)(EtOH)(-) or OsCl(5)(H(2)O)( ), neither of which is photocatalytically active. PMID- 20003155 TI - Hypericin hydroquinone: potential as a red-far red photosensitizer? AB - Hypericin hydroquinone is the product of two-electron reduction of hypericin (quinone), a potent phenanthroperylenequinone photosensitizer. In contrast to the quinone, the hydroquinone exhibits strong absorbance in the far-red spectral region. Herein we provide initial evidence on the potential of hypericin hydroquinone as a far-red photosensitizer. PMID- 20003156 TI - Exclusive observation of the (13R)-enantiomer of chlorophyll-c from a diatom Chaetoseros calcitrans. AB - Chiral high-performance liquid-chromatography (HPLC) for quantitative analysis of optically active chlorophyll(Chl)-c molecules, which are seen in many marine photosynthetic organisms, was developed. Chls-c have a single asymmetric carbon at the 13(2)-position, so their stereoisomers are (13(2)R)- and (13(2)S) enantiomers. After the separation of each enantiomer, the stereochemistry was unambiguously characterized using its circular dichroism spectrum in comparison with that of the structure-related compound, protochlorophyllide-a. Moreover, Chls-c were carefully extracted from the cells of a diatom Chaetoseros calcitrans without racemization and were subjected to the chiral HPLC. The results clearly demonstrated that naturally occurring Chl-c molecules are enantiomerically pure (13(2)R)-forms, which are generally found in photosynthetically active chlorophyllous pigments. PMID- 20003157 TI - Kinetics of the M-intermediate in the photocycle of bacteriorhodopsin upon chemical modification with surfactants. AB - The spectroscopic and kinetic studies of the interaction between bacteriorhodopsin in the M-intermediate and several surfactants (cetyl trimethyl ammonium bromide, dodecyl trimethyl ammonium bromide, diethylene glycol mono-n hexyl ether, ethylene glycol mono-n-hexyl ether, sodium 1-decanesulfonate and sodium 1-heptanesulfonate) have been investigated using steady-state UV-VIS spectrometry and time-resolved absorption techniques. The steady-state spectral results show that bR retains its trimeric state. Time-resolved observations indicate that the rate of deprotonation of the protonated Schiff base increases in the presence of the cationic surfactants, whereas insignificant changes are observed in the neutral or anionic surfactants. The rate of the reprotonation of the Schiff base in the transition M --> N is accelerated in anionic and neutral surfactants, but is decelerated in the presence of the cationic surfactants. Surfactants with a longer hydrocarbon tail have a greater effect on the kinetics when compared with surfactants having shorter hydrocarbon tails. The opposite effect is observed when the hydrophilic head of the surfactants contains opposite charges. These distinct kinetics are discussed in terms of the difference in the modified surface hydrophilicity of the bR and the possible protein configurational changes upon surfactant treatments. PMID- 20003158 TI - Fluorescent probing of urea-induced chemical unfolding of bovine serum albumin by intramolecular charge transfer fluorescence probe E-3-(4-dimethylamino-naphthalen 1-yl)-acrylic acid. AB - Changes in polarity at the immediate binding site in protein bovine serum albumin (BSA) produces distinct changes in the solvent polarity-dependent emission band of fluorescence probe E-3-(4-dimethylamino-naphthalen-1-yl)-acrylic acid. Steady state spectroscopy and time-resolved spectroscopy have been used to investigate this binding process. Attaching the probe to BSA and then monitoring its spectral changes with increasing urea concentration and raising temperature has also tracked the denaturation of BSA chemically and thermally. The polarity of the microenvironment was investigated employing the Reichardt E(T)(30) scale. Fluorescence anisotropy, red edge excitation shifts and acrylamide-induced quenching of fluorescence have been exploited to gain better insight into this binding process. PMID- 20003159 TI - Deferoxamine iron chelation increases delta-aminolevulinic acid induced protoporphyrin IX in xenograft glioma model. AB - Exogenous administration of delta-aminolevulinic acid (delta-ALA) leads to selective accumulation of protoporphyrin IX (PpIX) in brain tumors, and has shown promising results in increasing extent of resection in fluorescence-guided resection (FGR) of brain tumors. However, this approach still suffers from heterogeneous staining and so some tumor margins may go undetected because of this variation in PpIX production. The aim of this study was to test the hypothesis that iron chelation therapy could increase the level of fluorescence in malignant glioma tumors. Mice implanted with xenograft U251-GFP glioma tumor cells were given a 200 mg kg(-1) dose of deferoxamine (DFO), once a day for 3 days prior to delta-ALA administration. The PpIX fluorescence observed in the tumor regions was 1.9 times the background in animal group without DFO, and 2.9 times the background on average, in the DFO pre-treated group. A 50% increase in PpIX fluorescence contrast in the DFO group was observed relative to the control group (t-test P-value = 0.0020). These results indicate that iron chelation therapy could significantly increase delta-ALA-induced PpIX fluorescence in malignant gliomas, pointing to a potential role of iron chelation therapy for more effective FGR of brain tumors. PMID- 20003160 TI - Spectroscopic properties of fluorescein and rhodamine dyes attached to DNA. AB - We report the spectroscopic properties of fluorescein, x-rhodamine, tetramethyl rhodamine, attached to single strand, duplex DNA, and to the digestion products by DNAse I. The properties reported include: molar absorptivity, quantum yield, absorbance and fluorescence spectra, fluorescence lifetime, intrinsic lifetime (tau0), static quenching (S) and the Forster critical distances (R0) between fluorescein and x-rhodamine or tetramethyl-rhodamine (acceptors). These spectroscopic properties depend strongly on the local dye environment. Fluorescein was studied: (1) attached to biotin (BF), (2) BF bound to avidin; and attached to two positions in DNA. X-rhodamine and tetramethyl-rhodamine were studied as free dyes and attached at the 5'-end of DNA. We propose a general method to determine the molar absorptivity and tau0 of a dye attached to DNA based on the reaction of a biotinylated and dye-labeled oligomer with standardized avidin. The molar absorptivity of a second dye attached to a DNA duplex can be obtained by comparing spectra of doubly and singly labeled sequences. S, arising from dye-DNA interactions can then be determined. R0 for free and attached dyes showed differences from 1.1 to 4.2 A. We present evidence for the direct interaction of dyes attached to the termini of various single stranded DNA sequences. PMID- 20003161 TI - Childhood immunization and atopic disease into middle-age--a prospective cohort study. AB - The association between childhood immunizations and risk of atopic diseases is unclear. No study has examined possible associations between childhood immunizations and such diseases in middle age. The Tasmanian Longitudinal Health Study (TAHS) is a population based cohort study of respiratory disease. The TAHS participants were followed from 7 to 44 yrs of age. Immunizations during childhood were examined for any association with asthma and atopic disease at age 44 yrs. Multivariable regression models were used to estimate relative risks while adjusting for confounders. Cox regression was used to estimate the association between childhood immunizations and asthma developing after the age of 7 yrs. We found no association between any childhood immunization (Diphtheria, Tetanus, Pertussis, Polio, Smallpox) and asthma (ORs ranged from 0.87 to 1.17 p > 0.05), eczema (ORs ranged from 0.99 to 1.07 p > 0.05), food allergy (ORs ranged from 0.97 to 1.11 p > 0.05), or hay fever (ORs ranged from 1.02 to 1.05 p > 0.05) at age 44. Nor did we find any association between childhood immunizations and an increased risk of incident asthma after the age of 7 yrs (Diphtheria HR = 1.06, 95% CI 0.82, 1.36; Tetanus HR = 1.13, 95% CI 0.88, 1.44; Pertussis HR = 1.03, 95% CI 0.81, 1.30; Polio HR = 1.15, 95% CI 0.86, 1.54; Smallpox HR = 1.21, 95% CI 0.99, 1.48; DTP HR = 1.05, 95% CI 0.85, 1.30). Our analysis does not support any association between common childhood immunizations and risk of asthma and atopic disease in middle-age. Our findings should provide reassurance that in terms of life time risk of asthma and atopic disease, childhood immunization is safe. PMID- 20003162 TI - Suppressive effects of formoterol and salmeterol on eotaxin-1 in bronchial epithelial cells. AB - Eotaxin-1 (CCL11), an eosinophil-specific C-C chemokine, is a potent chemoattractant for mobilization of eosinophils into airways after allergic stimulation. Eotaxin-1 recruits eosinophils into inflammatory sites, and may play a role in the pathogenesis of asthma. Formoterol and salmeterol are two inhaled long acting beta(2) adrenoceptor agonists (LABAs), widely used for the local treatment of asthma. However, little is known about their effects on the eotaxin 1 expression of bronchial epithelial cells. BEAS-2B cells were stimulated by adding IL-4 with or without 2 h pre-treatment of formoterol or salmeterol. The protein and mRNA expression of eotaxin-1 were measured by ELISA assay and real time PCR, respectively. Effects of formoterol and salmeterol on nuclear and cytosolic pSTAT-6 expression were evaluated by Western blot and immunofluorescence study. Formoterol and salmeterol (10(-7)-10(-10) m) significantly down-regulated IL-4- induced eotaxin-1 expression in BEAS-2B cells. A specific beta(2) adrenoceptor antagonist (ICI 118,551) reversed their suppression of eotaxin-1 production. Forskolin, an cAMP activator, could also suppress the expression of eotaxin-1 by IL-4 in a dose dependent manner (10(-7) 10(-10 )m). The western blot and immunofluorescence studies demonstrated that formoterol 10(-7 )m suppressed the nuclear expression of pSTAT-6. Formoterol and salmeterol, two inhaled long-acting beta(2) agonists, down-regulated IL-4- induced eotaxin-1 expression in BEAS-2B cells. The effect was mediated via the beta(2) adrenoceptor, and cAMP. Formoterol significantly down-regulated pSTAT6 at higher concentration, and further turned off the IL-4 signaling pathway. PMID- 20003163 TI - Cat sensitization identified by recombinant Fel d 1 several years before symptoms -results from the BAMSE cohort. AB - Exposure to cat is one of the most important causes of allergic disease. The objective of this study was to investigate IgE reactivity to the recombinant major cat allergen, rFel d 1, as an early marker of cat sensitization. Based on questionnaires, 144 children with allergic symptoms due to cat, or where such symptoms were suspected, were selected from the birth cohort BAMSE and allocated into three study groups. Blood samples taken at age 4 and 8 yrs were analysed for IgE to rFel d 1 and cat dander extract (CDE) by quantitative ELISA (cut-off limit 0.037 kU(A)/l) and the ImmunoCAP System (cut-off limit 0.35 kU(A)/l), respectively. At 4 yrs, 25/33 children with certain allergic symptoms to cat had IgE to both rFel d 1 and CDE, while 14/42 of those suspecting symptoms at 4 had IgE to rFel d 1 and 9/42 to CDE. In a group developing symptoms after 4 yrs, 60/69 had IgE to rFel d 1 and 57/69 to CDE at 8, while 33/69 had IgE to rFel d 1 already at 4 and 26/69 to CDE. This was the only one of the three study groups where a significant increase in the IgE levels to rFel d 1 was found from 4 to 8 yrs (p < 0.001), even when only children with IgE to rFel d 1 already at 4 were included (p < 0.001). We show that the single major cat allergen rFel d 1 is at least as good as CDE in the diagnosis of cat allergy in childhood. With a sensitive rFel d 1 assay cat sensitization can be detected several years before symptoms to cat are reported. PMID- 20003164 TI - Functional conservation and diversification of class E floral homeotic genes in rice (Oryza sativa). AB - Mutant analyses in different eudicotyledonous flowering plants demonstrated that SEPALLATA-like MADS-box genes are required for the specification of sepals, petals, stamens and carpels, and for floral determinacy, thus defining class E floral organ identity genes. SEP-like genes encode MADS-domain transcription factors and constitute an angiosperm-specific gene clade whose members show remarkably different degrees of redundancy and sub-functionalization within eudicots. To better understand the evolutionary dynamics of SEP-like genes throughout the angiosperms we have knocked down SEP-like genes of rice (Oryza sativa), a distant relative of eudicots within the flowering plants. Plants affected in both OsMADS7 and OsMADS8 show severe phenotypes including late flowering, homeotic changes of lodicules, stamens and carpels into palea/lemma like organs, and a loss of floral determinacy. Simultaneous knockdown of the four rice SEP-like genes OsMADS1, OsMADS5, OsMADS7 and OsMADS8, leads to homeotic transformation of all floral organs except the lemma into leaf-like organs. This mimics the phenotype observed with the sep1 sep2 sep3 sep4 quadruple mutant of Arabidopsis. Detailed analyses of the spatial and temporal mRNA expression and protein interaction patterns corresponding to the different rice SEP-like genes show strong similarities, but also gene-specific differences. These findings reveal conservation of SEP-like genes in specifying floral determinacy and organ identities since the separation of eudicots and monocots about 150 million years ago. However, they indicate also monocot-specific neo- and sub-functionalization events and hence underscore the evolutionary dynamics of SEP-like genes. Moreover, our findings corroborate the view that the lodicules of grasses are homologous to eudicot petals. PMID- 20003165 TI - A dynamic gene expression atlas covering the entire life cycle of rice. AB - Growth and development of a plant are controlled by programmed expression of suits of genes at the appropriate time, tissue and abundance. Although genomic resources have been developed rapidly in recent years in rice, a model plant for cereal genome research, data of gene expression profiling are still insufficient to relate the developmental processes to transcriptomes, leaving a large gap between the genome sequence and phenotype. In this study, we generated genome wide expression data by hybridizing 190 Affymetrix GeneChip Rice Genome Arrays with RNA from 39 tissues collected throughout the life cycle of the rice plant from two varieties, Zhenshan 97 and Minghui 63. Analyses of the global transcriptomes revealed many interesting features of dynamic patterns of gene expression across the tissues and stages. In total, 38 793 probe sets were detected as expressed and 69% of the expressed transcripts showed significantly variable expression levels among tissues/organs. We found that similarity of transcriptomes among organs corresponded well to their developmental relatedness. About 5.2% of the expressed transcripts showed tissue-specific expression in one or both varieties and 22.7% of the transcripts exhibited constitutive expression including 19 genes with high and stable expression in all the tissues. This dataset provided a versatile resource for plant genomic research, which can be used for associating the transcriptomes to the developmental processes, understanding the regulatory network of these processes, tracing the expression profile of individual genes and identifying reference genes for quantitative expression analyses. PMID- 20003166 TI - Structure and evolutionary conservation of the plant N-end rule pathway. AB - The N-end rule relates the in vivo half-life of a protein to the identity of its N-terminal amino acid residue. While some N-terminal residues result in metabolically stable proteins, other, so-called destabilizing residues, lead to rapid protein turnover. The N-end rule pathway, which mediates the recognition and degradation of proteins with N-terminal destabilizing residues, is present in all organisms examined, including prokaryotes. This protein degradation pathway has a hierarchical organization in which some N-terminal residues, called primary destabilizing residues, are directly recognized by specific ubiquitin ligases. Other destabilizing residues, termed secondary and tertiary destabilizing residues, require modifications before the corresponding proteins can be targeted for degradation by ubiquitin ligases. In eukaryotes, the N-end rule pathway is a part of the ubiquitin/proteasome system and is known to play essential roles in a broad range of biological processes in fungi, animals and plants. While the structure of the N-end rule pathway has been extensively studied in yeast and mammals, knowledge of its organization in plants is limited. Using both tobacco and Arabidopsis, we identified the complete sets destabilizing and stabilizing N terminal residues. We also characterized the hierarchical organization of the plant N-end rule by identifying and determining the specificity of two distinct N terminal amidohydrolases (Nt-amidases) of Arabidopsis that are essential for the destabilizing activity of the tertiary destabilizing residues Asn and Gln. Our results indicate that both the N-end rule itself and mechanistic aspects of the N end rule pathway in angiosperms are very similar to those of mammals. PMID- 20003167 TI - Early transcriptomic events in microdissected Arabidopsis nematode-induced giant cells. AB - Root-knot nematodes differentiate highly specialized feeding cells in roots (giant cells, GCs), through poorly characterized mechanisms that include extensive transcriptional changes. While global transcriptome analyses have used galls, which are complex root structures that include GCs and surrounding tissues, no global gene expression changes specific to GCs have been described. We report on the differential transcriptome of GCs versus root vascular cells, induced in Arabidopsis by Meloidogyne javanica at a very early stage of their development, 3 days after infection (d.p.i.). Laser microdissection was used to capture GCs and root vascular cells for microarray analysis, which was validated through qPCR and by a promoter-GUS fusion study. Results show that by 3 d.p.i., GCs exhibit major gene repression. Although some genes showed similar regulation in both galls and GCs, the majority had different expression patterns, confirming the molecular distinctiveness of the GCs within the gall. Most of the differentially regulated genes in GCs have no previously assigned function. Comparisons with other transcriptome analyses revealed similarities between GCs and cell suspensions differentiating into xylem cells. This suggests a molecular link between GCs and developing vascular cells, which represent putative GC stem cells. Gene expression in GCs at 3 d.p.i. was also found to be similar to crown galls induced by Agrobacterium tumefaciens, a specialized root biotroph. PMID- 20003168 TI - Generation of pluripotent stem cells from eggs of aging mice. AB - Oocytes can reprogram genomes to form embryonic stem (ES) cells. Although ES cells largely escape senescence, oocytes themselves do senesce in the ovaries of most mammals. It remains to be determined whether ES cells can be established using eggs from old females, which exhibit reproductive senescence. We attempted to produce pluripotent stem cell lines from artificial activation of eggs (also called pES) from reproductive aged mice, to determine whether maternal aging affects pES cell production and pluripotency. We show that pES cell lines were generated with high efficiency from reproductive aged (old) mice, although parthenogenetic embryos from these mice produced fewer ES clones by initial two passages. Further, pES cell lines generated from old mice showed telomere length, expression of pluripotency molecular markers (Oct4, Nanog, SSEA1), alkaline phosphatase activity, teratoma formation and chimera production similar to young mice. Notably, DNA damage was reduced in pES cells from old mice compared to their progenitor parthenogenetic blastocysts, and did not differ from that of pES cells from young mice. Also, global gene expression differed only minimally between pES cells from young and old mice, in contrast to marked differences in gene expression in eggs from young and old mice. These data demonstrate that eggs from old mice can generate pluripotent stem cells, and suggest that the isolation and in vitro culture of ES cells must select cells with high levels of DNA and telomere integrity, and/or with capacity to repair DNA and telomeres. PMID- 20003169 TI - Live donor nephrectomy: a review of evidence for surgical techniques. AB - SUMMARY: Live kidney donation is an important alternative for patients with end stage renal disease. To date, the health of live kidney donors at long-term follow-up is good, and the procedure is considered to be safe. Surgical practice has evolved from the open lumbotomy, through mini-incision muscle-splitting open donor nephrectomy, to minimally invasive laparoscopic techniques. There are different minimally invasive techniques, including standard laparoscopic, hand assisted laparoscopic, hand-assisted retroperitoneoscopic, pure retroperitoneoscopic, and robotic-assisted live donor nephrectomy. At present, these minimally invasive techniques are subjected to clinical trials focusing on surgical outcome, quality of life, costs, long-term follow-up, and also morbidity of donor, recipient, and graft. In practice, many centers only perform donor nephrectomy on young healthy donors with normal weight. There is increasing evidence that donor nephrectomy with multiple arteries, right kidney and obese patients can be done with precaution. In this review, we address the surgical part of live kidney donation and the best level of evidence for all surgical techniques and issues surrounding the technique. PMID- 20003170 TI - Optimal placement of bipolar surface EMG electrodes in the face based on single motor unit analysis. AB - Locations of surface electromyography (sEMG) electrodes in the face are usually chosen on a macro-anatomical basis. In this study we describe optimal placement of bipolar electrodes based on a novel method and present results for lower facial muscles. We performed high-density sEMG recordings in 13 healthy participants. Raw sEMG signals were decomposed into motor unit action potentials (MUAPs). We positioned virtual electrode pairs in the interpolated monopolar MUAPs at different positions along muscle fiber direction and calculated the bipolar potentials. Electrode sites were determined where maximal bipolar amplitude was achieved and were validated. Objective guidelines for sEMG electrode placement improve the signal-to-noise ratio and may contribute to reduce cross talk, which is particularly important in the face. The method may be regarded as an important basis for improving the validity and reproducibility of sEMG in complex muscle areas. PMID- 20003172 TI - Flagellar motions in phototactic steering in a brown algal swarmer. AB - Using infrared high-speed video microscopy, we observed light-triggered transitory flagellar motions in flagellate reproductive cells (swarmers) of a brown alga, Scytosiphon lomentaria, under primary helical swimming conditions before and during negative phototactic orientation to unilateral actinic light. The posterior flagellum, which is autofluorescent and thought to be light sensing, was passively dragged in the dark and exhibited one to several rapid lateral beats during orientation changes for phototactic steering. Notably, a brief cessation of anterior flagellar beating was occasionally observed concomitantly with rapid beats of the posterior flagellum. This behavior caused a pause in helical body rotation, which may contribute to the accuracy of phototactic steering. Thus, coordinated regulation of the movement of the two flagella plays a crucial role in phototactic steering. PMID- 20003171 TI - Temporal requirements of insulin/IGF-1 signaling for proteotoxicity protection. AB - Toxic protein aggregation (proteotoxicity) is a unifying feature in the development of late-onset human neurodegenerative disorders. Reduction of insulin/IGF-1 signaling (IIS), a prominent lifespan, developmental and reproductive regulatory pathway, protects worms from proteotoxicity associated with the aggregation of the Alzheimer's disease-linked Abeta peptide. We utilized transgenic nematodes that express human Abeta and found that late life IIS reduction efficiently protects from Abeta toxicity without affecting development, reproduction or lifespan. To alleviate proteotoxic stress in the animal, the IIS requires heat shock factor (HSF)-1 to modulate a protein disaggregase, while DAF 16 regulates a presumptive active aggregase, raising the question of how these opposing activities could be co-regulated. One possibility is that HSF-1 and DAF 16 have distinct temporal requirements for protection from proteotoxicity. Using a conditional RNAi approach, we found an early requirement for HSF-1 that is distinct from the adult functions of DAF-16 for protection from proteotoxicity. Our data also indicate that late life IIS reduction can protect from proteotoxicity when it can no longer promote longevity, strengthening the prospect that IIS reduction might be a promising strategy for the treatment of neurodegenerative disorders caused by proteotoxicity. PMID- 20003173 TI - Effect of ultraviolet radiation on acetylcholinesterase activity in freshwater copepods. AB - We analyzed the effects of UV radiation (UVR) effects on acetylcholinesterase (AChE) activity in two calanoid copepods, Boeckella gibbosa and Parabroteas sarsi that inhabit Patagonian shallow lakes. We studied the effect of experimental UVR (UV-B and UV-A) exposure on AChE activity in relation to basal antioxidant capacities of both copepods. Our experiments showed that UVR can effectively depress AChE activity, although with differences between species. In both copepods AChE was affected by UV-B, whereas UV-A only affected AChE in B. gibbosa. Both copepods also differed in body elemental composition (C:N:P), photoprotecting compound content (carotenoids and mycosporine-like amino acids) and enzymatic antioxidant capacity (glutathione S-transferase [GST]). Our results suggest that when exposed to UVR, AChE activity would depend more on the antioxidant capacity (GST) and P availability for enzyme synthesis than on the photoprotective compounds. PMID- 20003174 TI - Photosensitizer-antibiotic conjugates: a novel class of antibacterial molecules. AB - Bacterial resistance to a variety of antibiotics has led to intensive research into the effect of photosensitizers as a cytotoxic agent against bacterial cells. In this study, we synthesized the following conjugates with or without a linker: rose bengal-penicillanic acid (RBPA), rose bengal-linker-penicillanic acid (RBLPA) and rose bengal-linker-kanamycin (RBLKAN). The antibacterial activity of these conjugates was examined on Staphylococcus aureus and Escherichia coli. Exposure of the cultures to 100 J cm(-2) showed that the minimum inhibitory concentration (MIC) of RBPA, RBLPA and RBLKAN on S. aureus was 0.195, 0.156 and 0.004 microm, respectively. The MIC of RBPA, RBLPA and RBLKAN on E. coli was 1.56, 2.5 and 0.156 microm, respectively. In dark control experiments, the MIC of these conjugates was not detected until a concentration that was 16-fold that of the MIC found in the light experiments. RBPA and RBLPA as well as RBLKAN are bactericidal for both bacterial cells. Total eradication of S. aureus and E. coli was observed with RBLKAN (0.078 and 20 microm 16 J cm(-2), respectively). Under these conditions, scanning electron microscopic analysis showed significant damage to these bacteria. However, the photosensitizer and antibiotics individually were not effective. PMID- 20003175 TI - Antibodies and DNA photoproducts: applications, milestones and reference guide. AB - Polyclonal and monoclonal antibodies recognizing the primary DNA photoproducts induced by ultraviolet radiation (UVR) have proven to be essential tools in the study of photochemical and photobiological phenomena. As specific "DNA damage binding proteins" these reagents have been used to develop a diverse array of technical procedures applied to a plethora of important problems in DNA photochemistry and the biological effects of UVR at the molecular, developmental, organism and population levels. This survey attempts to cover this science from an historical perspective and to reveal the great breadth of discovery and contribution associated with the development and application of DNA damage antibodies to the current body of science. PMID- 20003176 TI - Opiate-induced oesophageal dysmotility. AB - BACKGROUND: Opiates have well characterized (troublesome) untoward effects on the gastrointestinal tract. Opioid bowel dysfunction has been a subject of research and even drug design, but surprisingly little is known with regard to clinical effects of opiates on the oesophagus. AIM: To characterize opiate effects on motor function of the oesophagus in patients presenting with dysphagia. METHODS: Retrospective review of 15 patients with dysphagia referred for oesophageal manometry while on chronic opiates. Manometry was completed during opiate use and in three cases, after opiates were discontinued. RESULTS: All patients demonstrated motility abnormalities. Incomplete lower oesophageal sphincter (LOS) relaxation (11.5 +/- 1.6 mmHg) was seen in most cases. Ten patients demonstrated nonperistaltic contractions in > or =3 of 10 swallows. Additional abnormalities included high amplitude contractions; triple peaked contractions; and increased velocity. The average resting lower oesophageal sphincter (LOSP) met criteria for hypertensive LOS in three patients. These features were suggestive of spasm or achalasia. Repeat manometry off opiates was performed in three cases. LOS relaxation was noted to be complete upon repeat manometry in these cases. There was also improved peristalsis and normal velocity. CONCLUSIONS: A range of manometric abnormalities were seen in patients with dysphagia in the setting of opiate use: impaired LOS relaxation, high amplitude/velocity and simultaneous oesophageal waves. These data suggest that the oesophagus is susceptible to the effects of opiates and care must be taken before ascribing dysphagia to a primary oesophageal motility disorder in patients taking opiates. PMID- 20003177 TI - What we still don't know about AIDS. PMID- 20003178 TI - Malaria paediatric hospitalization between 1999 and 2008 across Kenya. AB - BACKGROUND: Intervention coverage and funding for the control of malaria in Africa has increased in recent years, however, there are few descriptions of changing disease burden and the few reports available are from isolated, single site observations or are of reports at country-level. Here we present a nationwide assessment of changes over 10 years in paediatric malaria hospitalization across Kenya. METHODS: Paediatric admission data on malaria and non-malaria diagnoses were assembled for the period 1999 to 2008 from in-patient registers at 17 district hospitals in Kenya and represented the diverse malaria ecology of the country. These data were then analysed using autoregressive moving average time series models with malaria and all-cause admissions as the main outcomes adjusted for rainfall, changes in service use and populations-at-risk within each hospital's catchment to establish whether there has been a statistically significant decline in paediatric malaria hospitalization during the observation period. RESULTS: Among the 17 hospital sites, adjusted paediatric malaria admissions had significantly declined at 10 hospitals over 10 years since 1999; had significantly increased at four hospitals, and remained unchanged in three hospitals. The overall estimated average reduction in malaria admission rates was 0.0063 cases per 1,000 children aged 0 to 14 years per month representing an average percentage reduction of 49% across the 10 hospitals registering a significant decline by the end of 2008. Paediatric admissions for all-causes had declined significantly with a reduction in admission rates of greater than 0.0050 cases per 1,000 children aged 0 to 14 years per month at 6 of 17 hospitals. Where malaria admissions had increased three of the four sites were located in Western Kenya close to Lake Victoria. Conversely there was an indication that areas with the largest declines in malaria admission rates were areas located along the Kenyan coast and some sites in the highlands of Kenya. CONCLUSION: A country-wide assessment of trends in malaria hospitalizations indicates that all is not equal, important variations exist in the temporal pattern of malaria admissions between sites and these differences require more detailed investigation to understand what is required to promote a clinical transition across Africa. PMID- 20003179 TI - Genetic variation of NEDD4L is associated with essential hypertension in female Kazakh general population: a case-control study. AB - BACKGROUND: Hypertension affects > 18.8% of adults in China. Indeed, hypertension is the most prevalent risk factor for cardiovascular morbidity and mortality worldwide. Genetic variation is thought to contribute to the etiology of hypertension. NEDD4L is a candidate gene for hypertension, both functionally and genetically. The purpose of the current study was to investigate the relationship between the variation in NEDD4L and essential hypertension in Kazakh, which is a relatively isolated population with a pure genetic background and is an ideal population to study genetic mechanisms of hypertension. METHODS: We screened the promoter and exons of NEDD4L in 94 Kazakh hypertensive individuals to identify representative variations. Then, by genotyping the representative variations in the Kazakh general population, a case-control study was conducted. RESULTS: By systemically screening variations of NEDD4L, we did not identify any functional mutations in NEDD4L. A new common variation (296921-296923delTTG), which is not found in the NCBI database, was identified. Three representative variations (296921-296923delTTG, rs2288774, and rs2288775) were successfully genotyped in the Kazakh general population. The distribution of the dominant model (AA vs. AG+GG) of rs2288775, the additive model, and the recessive model (II+ID vs. DD) of 296921-296923delTTG differed significantly between the cases and controls in females (P = 0.040, P = 0.024, and P = 0.007, respectively). After adjusting for confounding factors, logistic regression analysis showed that rs2288775 (in the dominant model) and 296921-296923delTTG (in the recessive model) were significantly associated with hypertension (rs2288775: OR = 1.479, 95% CI = 1.011 2.064, p = 0.044; and 296921-296923delTTG: OR = 1.908, 95% CI = 1.020-3.568, p = 0.043) in females. The frequency of the D-C-G haplotype was significantly higher for cases than for controls in females (P = 0.020). There was a significant interaction between the NEDD4L genotype and gender (P for interaction: 0.045 for rs2288775 and 0.064 for 296921-296923delTTG), but there was no significant interaction between the NEDD4L genotype and smoking (P for interaction: 0.616 for rs2288775 and 0.447 for 296921-296923delTTG). For females and total participants, the urinary Na excretion rate was significantly lower in the DD than the I/I+I/D individuals (P = 0.032 and P = 0.027 respectively). CONCLUSION: The genetic variations of NEDD4L may be associated with essential hypertension in females in the Kazakh general population. PMID- 20003180 TI - Eradication of intracellular Francisella tularensis in THP-1 human macrophages with a novel autophagy inducing agent. AB - BACKGROUND: Autophagy has been shown recently to play an important role in the intracellular survival of several pathogenic bacteria. In this study, we investigated the effect of a novel small-molecule autophagy-inducing agent, AR 12, on the survival of Francisella tularensis, the causative bacterium of tularemia in humans and a potential bioterrorism agent, in macrophages. METHODS AND RESULTS: Our results show that AR-12 induces autophagy in THP-1 macrophages, as indicated by increased autophagosome formation, and potently inhibits the intracellular survival of F. tularensis (type A strain, Schu S4) and F. novicida in macrophages in association with increased bacterial co-localization with autophagosomes. The effect of AR-12 on intracellular F. novicida was fully reversed in the presence of the autophagy inhibitor, 3-methyl adenine or the lysosome inhibitor, chloroquine. Intracellular F. novicida were not susceptible to the inhibitory activity of AR-12 added at 12 h post-infection in THP-1 macrophages, and this lack of susceptibility was independent of the intracellular location of bacteria. CONCLUSION: Together, AR-12 represents a proof-of-principle that intracellular F. tularensis can be eradicated by small-molecule agents that target innate immunity. PMID- 20003181 TI - Mortality effects of average education: a multilevel study of small neighbourhoods in rural and urban areas in Norway. AB - BACKGROUND: The intention was to find out whether there was an association between the socio-economic resources in a small neighbourhood ("basic statistical unit"; BSU) and individual mortality, net of individual resources, and whether this association differed between municipalities including a quite large city and others. The possibility of a rural-urban difference in the health effect of community resources has not been checked earlier. METHODS: Discrete-time hazard models for mortality at age 60-89 were estimated for 1990-1992 and 2000-2002, using register data that cover the entire Norwegian population. For each person, the educational level and the municipality and BSU of residence in 1990 and 2000 were known. Average education was computed by aggregating over the individual data. In total, there were about 200000 deaths in more than 13000 BSUs during 5 million person-years of observation. RESULTS: There was a significant relationship between average education in the BSU and individual mortality, but only in the medium-sized and largest municipalities. The sharpest relationship was seen in the latter, where for example OR per year of education was 0.908 (95% CI 0.887-0.929) in the 1990-92 period. The findings were robust to various alternative specifications. CONCLUSION: These results from a large data set are consistent with the idea that neighbourhood socio-economic resources may affect individual mortality, but suggest that distinctions according to population size or density be made in future research and that one should be careful, if focusing on cities, to generalize beyond that setting. With these data, one can only speculate about the reasons for the rural-urban difference. A stronger higher level spatial segregation in urban areas may be one explanation. PMID- 20003182 TI - The hydrogenosomes of Psalteriomonas lanterna. AB - BACKGROUND: Hydrogenosomes are organelles that produce molecular hydrogen and ATP. The broad phylogenetic distribution of their hosts suggests that the hydrogenosomes of these organisms evolved several times independently from the mitochondria of aerobic progenitors. Morphology and 18S rRNA phylogeny suggest that the microaerophilic amoeboflagellate Psalteriomonas lanterna, which possesses hydrogenosomes and elusive "modified mitochondria", belongs to the Heterolobosea, a taxon that consists predominantly of aerobic, mitochondriate organisms. This taxon is rather unrelated to taxa with hitherto studied hydrogenosomes. RESULTS: Electron microscopy of P. lanterna flagellates reveals a large globule in the centre of the cell that is build up from stacks of some 20 individual hydrogenosomes. The individual hydrogenosomes are surrounded by a double membrane that encloses a homogeneous, dark staining matrix lacking cristae. The "modified mitochondria" are found in the cytoplasm of the cell and are surrounded by 1-2 cisterns of rough endoplasmatic reticulum, just as the mitochondria of certain related aerobic Heterolobosea. The ultrastructure of the "modified mitochondria" and hydrogenosomes is very similar, and they have the same size distribution as the hydrogenosomes that form the central stack.The phylogenetic analysis of selected EST sequences (Hsp60, Propionyl-CoA carboxylase) supports the phylogenetic position of P. lanterna close to aerobic Heterolobosea (Naegleria gruberi). Moreover, this analysis also confirms the identity of several mitochondrial or hydrogenosomal key-genes encoding proteins such as a Hsp60, a pyruvate:ferredoxin oxidoreductase, a putative ADP/ATP carrier, a mitochondrial complex I subunit (51 KDa), and a [FeFe] hydrogenase. CONCLUSION: Comparison of the ultrastructure of the "modified mitochondria" and hydrogenosomes strongly suggests that both organelles are just two morphs of the same organelle. The EST studies suggest that the hydrogenosomes of P. lanterna are physiologically similar to the hydrogenosomes of Trichomonas vaginalis and Trimastix pyriformis. Phylogenetic analysis of the ESTs confirms the relationship of P. lanterna with its aerobic relative, the heterolobosean amoeboflagellate Naegleria gruberi, corroborating the evolution of hydrogenosomes from a common, mitochondriate ancestor. PMID- 20003183 TI - Risk factors for early recurrence after inguinal hernia repair. AB - BACKGROUND: Family history, male gender and age are significant risk factors for inguinal hernia disease. Family history provides evidence for a genetic trait and could explain early recurrence after inguinal hernia repair despite technical advance at least in a subgroup of patients. This study evaluates if age and family history can be identified as risk factors for early recurrence after primary hernia repair. METHODS: We performed an observational cohort study for 75 patients having at least two recurrent hernias. The impact of age, gender and family history on the onset of primary hernias, age at first recurrence and recurrence rates was investigated. RESULTS: 44% (33/75) of recurrent hernia patients had a family history and primary as well as recurrent hernias occurred significantly earlier in this group (p = 0.04). The older the patients were at onset the earlier they got a recurrent hernia. Smoking could be identified as on additional risk factor for early onset of hernia disease but not for hernia recurrence. CONCLUSION: Our data reveal an increased incidence of family history for recurrent hernia patients when compared with primary hernia patients. Patients with a family history have their primary hernias as well as their recurrence at younger age then patients without a family history. Though recurrent hernia has to be regarded as a disease caused by multiple factors, a family history may be considered as a criterion to identify the risk for recurrence before the primary operation. PMID- 20003184 TI - Development of multiplex real-time PCR assays for identification of members of the Anopheles funestus species group. AB - BACKGROUND: The malaria vector and non-vector species of the Anopheles funestus group are morphologically very similar and accurate identification is required as part of effective control strategies. In the past, this has relied on morphological and cytogenetic methods but these have been largely superseded by a robust allele-specific PCR (AS-PCR). One disadvantage of AS-PCR is the requirement for post-PCR processing by gel electrophoresis of PCR products. In this study, three new high-throughput 'closed-tube' assays were developed and compared with the previously described AS-PCR technique. METHODS: Protocols for three fluorescence-based assays based on Melt Curve Analysis (MCA), High Resolution Melt (HRM) and TaqMan SNP genotyping were developed to detect and discriminate Anopheles parensis, Anopheles leesoni, Anopheles vaneedeni, Anopheles rivulorum and An. funestus s.s. The sensitivity and specificity of these assays were compared with the widely used AS-PCR in a blind trial using DNA extracted from wild-caught mosquitoes. RESULTS: The TaqMan assay proved to be the most sensitive and specific of the three new assays. The MCA and HRM assays initially gave promising results, but were more sensitive to both DNA quality and quantity and consequently showed a higher rate of incorrect identifications. CONCLUSION: The TaqMan assay proved to be the most robust of the three protocols tested in this study. This assay very effectively identified all five members of the An. funestus group using fluorescently-labeled probes with distinct emission and excitation spectra allowing their independent detection in a single reaction. This method is at least as sensitive and specific as the gold standard AS-PCR approach and because it has no requirement for post-PCR processing is simpler and more rapid to run. The one disadvantage of the TaqMan assay is the cost of this assay, both in terms of initial capital outlay and running cost per sample, which is higher than AS-PCR. However, the cost of both the real-time PCR machine and fluorescently labelled probes required is falling and in the future the cost of this assay is likely to become closer to that of standard PCR. PMID- 20003185 TI - Gene doctoring: a method for recombineering in laboratory and pathogenic Escherichia coli strains. AB - BACKGROUND: Homologous recombination mediated by the lambda-Red genes is a common method for making chromosomal modifications in Escherichia coli. Several protocols have been developed that differ in the mechanisms by which DNA, carrying regions homologous to the chromosome, are delivered into the cell. A common technique is to electroporate linear DNA fragments into cells. Alternatively, DNA fragments are generated in vivo by digestion of a donor plasmid with a nuclease that does not cleave the host genome. In both cases the lambda-Red gene products recombine homologous regions carried on the linear DNA fragments with the chromosome. We have successfully used both techniques to generate chromosomal mutations in E. coli K-12 strains. However, we have had limited success with these lambda-Red based recombination techniques in pathogenic E. coli strains, which has led us to develop an enhanced protocol for recombineering in such strains. RESULTS: Our goal was to develop a high throughput recombineering system, primarily for the coupling of genes to epitope tags, which could also be used for deletion of genes in both pathogenic and K-12 E. coli strains. To that end we have designed a series of donor plasmids for use with the lambda-Red recombination system, which when cleaved in vivo by the I SceI meganuclease generate a discrete linear DNA fragment, allowing for C terminal tagging of chromosomal genes with a 6xHis, 3xFLAG, 4xProteinA or GFP tag or for the deletion of chromosomal regions. We have enhanced existing protocols and technologies by inclusion of a cassette conferring kanamycin resistance and, crucially, by including the sacB gene on the donor plasmid, so that all but true recombinants are counter-selected on kanamycin and sucrose containing media, thus eliminating the need for extensive screening. This method has the added advantage of limiting the exposure of cells to the potential damaging effects of the lambda Red system, which can lead to unwanted secondary alterations to the chromosome. CONCLUSION: We have developed a counter-selective recombineering technique for epitope tagging or for deleting genes in E. coli. We have demonstrated the versatility of the technique by modifying the chromosome of the enterohaemorrhagic O157:H7 (EHEC), uropathogenic CFT073 (UPEC), enteroaggregative O42 (EAEC) and enterotoxigenic H10407 (ETEC) E. coli strains as well as in K-12 laboratory strains. PMID- 20003186 TI - Inactivation of the fliY gene encoding a flagellar motor switch protein attenuates mobility and virulence of Leptospira interrogans strain Lai. AB - BACKGROUND: Pathogenic Leptospira species cause leptospirosis, a zoonotic disease of global importance. The spirochete displays active rotative mobility which may contribute to invasion and diffusion of the pathogen in hosts. FliY is a flagellar motor switch protein that controls flagellar motor direction in other microbes, but its role in Leptospira, and paricularly in pathogenicity remains unknown. RESULTS: A suicide plasmid for the fliY gene of Leptospira interrogans serogroup Icterohaemorrhagiae serovar Lai strain Lai that was disrupted by inserting the ampicillin resistance gene (bla) was constructed, and the inactivation of fliY gene in a mutant (fliY-) was confirmed by PCR and Western Blot analysis. The inactivation resulted in the mRNA absence of fliP and fliQ genes which are located downstream of the fliY gene in the same operon. The mutant displayed visibly weakened rotative motion in liquid medium and its migration on semisolid medium was also markedly attenuated compared to the wild type strain. Compared to the wild-type strain, the mutant showed much lower levels of adhesion to murine macrophages and apoptosis-inducing ability, and its lethality to guinea pigs was also significantly decreased. CONCLUSION: Inactivation of fliY, by the method used in this paper, clearly had polar effects on downstream genes. The phentotypes observed, including lower pathogenicity, could be a consequence of fliY inactivation, but also a consequence of the polar effects. PMID- 20003187 TI - The effect on the small bowel of 5-FU and oxaliplatin in combination with radiation using a microcolony survival assay. AB - BACKGROUND: In locally advanced rectal cancer, 5-Fluorouracil (5-FU)-based chemoradiation is the standard treatment. The main acute toxicity of this treatment is enteritis. Due to its potential radiosensitizing properties, oxaliplatin has recently been incorporated in many clinical chemoradiation protocols. The aim of this study was to investigate to what extent 5-FU and oxaliplatin influence the radiation (RT) induced small bowel mucosal damage when given in conjunction with single or split dose RT. METHODS: Immune competent balb c mice were treated with varying doses of 5-FU, oxaliplatin (given intraperitoneally) and total body RT, alone or in different combinations in a series of experiments. The small bowel damage was studied by a microcolony survival assay. The treatment effect was evaluated using the inverse of the slope (D0) of the exponential part of the dose-response curve. RESULTS: In two separate experiments the dose-response relations were determined for single doses of RT alone, yielding D0 values of 2.79 Gy (95% CI: 2.65 - 2.95) and 2.98 Gy (2.66 - 3.39), for doses in the intervals of 5-17 Gy and 5-10 Gy, respectively. Equitoxic low doses (IC5) of the two drugs in combination with RT caused a decrease in jejunal crypt count with significantly lower D0: 2.30 Gy (2.10 - 2.56) for RT+5 FU and 2.27 Gy (2.08 - 2.49) for RT+oxaliplatin. Adding both drugs to RT did not further decrease D0: 2.28 Gy (1.97 - 2.71) for RT+5-FU+oxaliplatin. A clearly higher crypt survival was noted for split course radiation (3 x 2.5 Gy) compared to a single fraction of 7.5 Gy. The same difference was seen when 5-FU and/or oxaliplatin were added. CONCLUSION: Combining 5-FU or oxaliplatin with RT lead to an increase in mucosal damage as compared to RT alone in our experimental setting. No additional reduction of jejunal crypt counts was noted when both drugs were combined with single dose RT. The higher crypt survival with split dose radiation indicates a substantial recovery between radiation fractions. This mucosal-sparing effect achieved by fractionation was maintained also when chemotherapy was added. PMID- 20003188 TI - Evaluating the financial protection of patients with chronic disease by health insurance in rural China. AB - BACKGROUND: A growing number of developing countries are developing health insurance schemes that aim to protect households, particularly the poor, from financial catastrophe and impoverishment caused by unaffordable medical care. This paper investigates the extent to which patients suffering from chronic disease in rural China face catastrophic expenditure on healthcare, and how far the New Co-operative Medical Insurance Scheme (NCMS) offers them financial protection against this. METHODS: A household survey was conducted in six counties in Ningxia Autonomous Region and Shandong Province, with a total of 6,147 rural households, including 3944 individual chronic disease patients. Structured questionnaires were used with chronic disease patients to investigate: their basic social and economic characteristics, including income and expenditure levels and NCMS membership; and their health care utilization, associated healthcare costs and levels of reimbursement by NCMS. 'Catastrophic' expenditure was defined as healthcare expenditure of more than 40% of household non-food expenditure. RESULTS: Expenditure for chronic diseases accounted for an average of 27% of annual non-food per capita expenditure amongst NCMS members in Shandong and 35% in Ningxia. 14-15% of families in both provinces spent more than 40% of their non-food expenditure on chronic healthcare costs. Between 8 and 11% of non NCMS members and 13% of NCMS members did not seek any medical care for chronic illness. A greater proportion of NCMS members in the poorest quintile faced catastrophic expenditure as compared to those in the richest quintile in both study sites. A slightly higher proportion of non-NCMS members than NCMS member households faced catastrophic expenditure, but the difference was not statistically significant. CONCLUSION: A significant proportion of patients with chronic diseases face catastrophic healthcare costs and these are especially heavy for the poor. The NCMS offers only a limited degree of financial protection. The heavy financial burden of healthcare for chronic disease poses an urgent challenge to the NCMS. There is an urgent need for a clear policy on how to offer financial protection to those with chronic disease. PMID- 20003189 TI - The effects of zooprophylaxis and other mosquito control measures against malaria in Nouna, Burkina Faso. AB - BACKGROUND: In the absence of large scale, organized vector control programmes, individual protective measures against mosquitoes are essential for reducing the transmission of diseases like malaria. Knowledge of the types and effectiveness of mosquito control methods used by households can aid in the development and promotion of preventive measures. METHODS: A matched, population-based case control study was carried out in the semi-urban region of Nouna, Burkina Faso. Surveys and mosquito captures were conducted for each participating household. Data were analysed using conditional logistic regression and Pearson's product moment correlations. RESULTS: In Nouna, Burkina Faso, the main types of reported mosquito control measures used included sleeping under bed nets (insecticide treated and untreated) and burning mosquito coils. Most of the study households kept animals within the compound or house at night. Insecticide house sprays, donkeys, rabbits and pigs were significantly associated with a reduced risk of malaria only in univariate analyses. CONCLUSION: Given the conflicting results of the effects of zooprophylaxis from previous studies, other community-based preventive measures, such as bed nets, coils and insecticide house-spraying, may be of more benefit. PMID- 20003190 TI - Estimation of the diameter and cross-sectional area of the internal jugular veins in adult patients. AB - INTRODUCTION: Unawareness of an asymmetry between the right and left internal jugular vein (IJV) and methodological pitfalls in previous studies raise concerns about such asymmetry. Hence the aim of this prospective non-interventional study was to validate the hypothesis that right IJV diameter is greater than those of left IJV and to determine the cross-sectional area of the IJVs using computed tomography (CT)-scans and original automatic software. METHODS: All consecutive adult outpatients who underwent a thoracic contrast-enhanced (TCE) helical CT scan during a 5-month period were included. To determine diameter and cross sectional area of the IJVs, we used Advanced Vessel Analysis software integrated in a CT-scan (Advanced Vessel Analysis on Advantage Workstation Windows 4.2; General Electrics) allowing automatic segmentation of vessels and calculation of their diameters and cross-sectional areas. RESULTS: A total of 360 TCE CT-scans was performed; 170 were excluded from the analysis. On the remaining 190 CT scans, the diameter and cross-sectional area of the right IJV were significantly greater than those of the left IJV (17 +/- 5 mm [median: 17 mm, range: 13 to 20 mm] vs. 14 +/- 5 mm [median: 13 mm, range: 10 to 16 mm], P < 0.001; and 181 +/- 111 mm2 [median: 160 mm2, range: 108 to 235 mm2] vs. 120 +/- 81 mm2 [median: 102 mm2, range: 63 to 168 mm2], P < 0.001, respectively). CONCLUSIONS: In a general population of adult outpatients, the diameter and cross-sectional area of the right IJV were significantly greater than those of the left IJV. This could be an additional argument to prefer right over left IJV cannulation. PMID- 20003191 TI - Identification of small molecule compounds that inhibit the HIF-1 signaling pathway. AB - BACKGROUND: Hypoxia-inducible factor-1 (HIF-1) is the major hypoxia-regulated transcription factor that regulates cellular responses to low oxygen environments. HIF-1 is composed of two subunits: hypoxia-inducible HIF-1alpha and constitutively-expressed HIF-1beta. During hypoxic conditions, HIF-1alpha heterodimerizes with HIF-1beta and translocates to the nucleus where the HIF-1 complex binds to the hypoxia-response element (HRE) and activates expression of target genes implicated in cell growth and survival. HIF-1alpha protein expression is elevated in many solid tumors, including those of the cervix and brain, where cells that are the greatest distance from blood vessels, and therefore the most hypoxic, express the highest levels of HIF-1alpha. Therapeutic blockade of the HIF-1 signaling pathway in cancer cells therefore provides an attractive strategy for development of anticancer drugs. To identify small molecule inhibitors of the HIF-1 pathway, we have developed a cell-based reporter gene assay and screened a large compound library by using a quantitative high throughput screening (qHTS) approach. RESULTS: The assay is based upon a beta lactamase reporter under the control of a HRE. We have screened approximate 73,000 compounds by qHTS, with each compound tested over a range of seven to fifteen concentrations. After qHTS we have rapidly identified three novel structural series of HIF-1 pathway Inhibitors. Selected compounds in these series were also confirmed as inhibitors in a HRE beta-lactamase reporter gene assay induced by low oxygen and in a VEGF secretion assay. Three of the four selected compounds tested showed significant inhibition of hypoxia-induced HIF-1alpha accumulation by western blot analysis. CONCLUSION: The use of beta-lactamase reporter gene assays, in combination with qHTS, enabled the rapid identification and prioritization of inhibitors specific to the hypoxia induced signaling pathway. PMID- 20003192 TI - Inhibition of trypsin expression in Lutzomyia longipalpis using RNAi enhances the survival of Leishmania. AB - BACKGROUND: Leishmania parasites must overcome several barriers to achieve transmission by their sand fly vectors. One of the earliest threats is exposure to enzymes during blood meal digestion. Trypsin-like enzymes appear to be detrimental to parasite survival during the very early phase of development as amastigotes transform into promastigote stages. Here, we investigate whether parasites can affect trypsin secretion by the sand fly midgut epithelium and if inhibition of this process is of survival value to the parasites. RESULTS: Infections of Lutzomyia longipalpis with Leishmania mexicana were studied and these showed that infected sand flies produced less trypsin-like enzyme activity during blood meal digestion when compared to uninfected controls. RNA interference was used to inhibit trypsin 1 gene expression by micro-injection into the thorax, as trypsin 1 is the major blood meal induced trypsin activity in the sand fly midgut. Injection of specific double stranded RNA reduced trypsin 1 expression as assessed by RT-PCR and enzyme assays, and also led to increased numbers of parasites in comparison with mock-injected controls. Injection by itself was observed to have an inhibitory effect on the level of infection, possibly through stimulation of a wound repair or immune response by the sand fly. CONCLUSION: Leishmania mexicana was shown to be able to modulate trypsin secretion by Lutzomyia longipalpis to its own advantage, and direct inhibition of trypsin gene expression led to increased parasite numbers in the midguts of infected flies. Successful application of RNA interference methodology to Leishmania-infected sand flies now opens up the use of this technique to study a wide range of sand fly genes and their role in the parasite-vector interaction. PMID- 20003193 TI - Genome-based polymorphic microsatellite development and validation in the mosquito Aedes aegypti and application to population genetics in Haiti. AB - BACKGROUND: Microsatellite markers have proven useful in genetic studies in many organisms, yet microsatellite-based studies of the dengue and yellow fever vector mosquito Aedes aegypti have been limited by the number of assayable and polymorphic loci available, despite multiple independent efforts to identify them. Here we present strategies for efficient identification and development of useful microsatellites with broad coverage across the Aedes aegypti genome, development of multiplex-ready PCR groups of microsatellite loci, and validation of their utility for population analysis with field collections from Haiti. RESULTS: From 79 putative microsatellite loci representing 31 motifs identified in 42 whole genome sequence supercontig assemblies in the Aedes aegypti genome, 33 microsatellites providing genome-wide coverage amplified as single copy sequences in four lab strains, with a range of 2-6 alleles per locus. The tri nucleotide motifs represented the majority (51%) of the polymorphic single copy loci, and none of these was located within a putative open reading frame. Seven groups of 4-5 microsatellite loci each were developed for multiplex-ready PCR. Four multiplex-ready groups were used to investigate population genetics of Aedes aegypti populations sampled in Haiti. Of the 23 loci represented in these groups, 20 were polymorphic with a range of 3-24 alleles per locus (mean = 8.75). Allelic polymorphic information content varied from 0.171 to 0.867 (mean = 0.545). Most loci met Hardy-Weinberg expectations across populations and pairwise FST comparisons identified significant genetic differentiation between some populations. No evidence for genetic isolation by distance was observed. CONCLUSION: Despite limited success in previous reports, we demonstrate that the Aedes aegypti genome is well-populated with single copy, polymorphic microsatellite loci that can be uncovered using the strategy developed here for rapid and efficient screening of genome supercontig assemblies. These loci are suitable for genetic and population studies using multiplex-PCR. PMID- 20003195 TI - Seven mistakes and potential solutions in epidemiology, including a call for a World Council of Epidemiology and Causality. AB - All sciences make mistakes, and epidemiology is no exception. I have chosen 7 illustrative mistakes and derived 7 solutions to avoid them. The mistakes (Roman numerals denoting solutions) are: 1. Failing to provide the context and definitions of study populations. (I Describe the study population in detail) 2. Insufficient attention to evaluation of error. (II Don't pretend error does not exist.) 3. Not demonstrating comparisons are like-for-like. (III Start with detailed comparisons of groups.) 4. Either overstatement or understatement of the case for causality. (IV Never say this design cannot contribute to causality or imply causality is ensured by your design.) 5. Not providing both absolute and relative summary measures. (V Give numbers, rates and comparative measures, and adjust summary measures such as odds ratios appropriately.) 6. In intervention studies not demonstrating general health benefits. (VI Ensure general benefits (mortality/morbidity) before recommending application of cause-specific findings.) 7. Failure to utilise study data to benefit populations. (VII Establish a World Council on Epidemiology to help infer causality from associations and apply the work internationally.) Analysis of these and other common mistakes is needed to benefit from the increasing discovery of associations that will be multiplying as data mining, linkage, and large-scale scale epidemiology become commonplace. PMID- 20003194 TI - Regurgitation in healthy and non healthy infants. AB - Uncomplicate regurgitation in otherwise healthy infants is not a disease. It consists of milk flow from mouth during or after feeding. Common causes include overfeeding, air swallowed during feeding, crying or coughing; physical exam is normal and weight gain is adequate. History and physical exam are diagnostic, and conservative therapy is recommended. Pathologic gastroesophageal reflux or gastroesophageal reflux disease refers to infants with regurgitation and vomiting associated with poor weight gain, respiratory symptoms, esophagitis. Reflux episodes occur most often during transient relaxations of the lower esophageal sphincter unaccompanied by swallowing, which permit gastric content to flow into the esophagus. A minor proportion of reflux episodes occurs when the lower esophageal sphincter fails to increase pressure during a sudden increase in intraabdominal pressure or when lower esophageal sphincter resting pressure is chronically reduced. Alterations in several protective mechanisms allow physiologic reflux to become gastroesophageal reflux disease; diagnostic approach is both clinical and instrumental: radiological series are useful to exclude anatomic abnormalities; pH-testing evaluates the quantity, frequency and duration of the acid reflux episodes; endoscopy and biopsy are performed in the case of esophagitis. Therapy with H2 receptor antagonists and proton pump inhibitors are suggested. PMID- 20003196 TI - The mitochondrial genomes of sponges provide evidence for multiple invasions by Repetitive Hairpin-forming Elements (RHE). AB - BACKGROUND: The mitochondrial (mt) genomes of sponges possess a variety of features, which appear to be intermediate between those of Eumetazoa and non metazoan opisthokonts. Among these features is the presence of long intergenic regions, which are common in other eukaryotes, but generally absent in Eumetazoa. Here we analyse poriferan mitochondrial intergenic regions, paying particular attention to repetitive sequences within them. In this context we introduce the mitochondrial genome of Ircinia strobilina (Lamarck, 1816; Demospongiae: Dictyoceratida) and compare it with mtDNA of other sponges. RESULTS: Mt genomes of dictyoceratid sponges are identical in gene order and content but display major differences in size and organization of intergenic regions. An even higher degree of diversity in the structure of intergenic regions was found among different orders of demosponges. One interesting observation made from such comparisons was of what appears to be recurrent invasions of sponge mitochondrial genomes by repetitive hairpin-forming elements, which cause large genome size differences even among closely related taxa. These repetitive hairpin-forming elements are structurally and compositionally divergent and display a scattered distribution throughout various groups of demosponges. CONCLUSION: Large intergenic regions of poriferan mt genomes are targets for insertions of repetitive hairpin- forming elements, similar to the ones found in non-metazoan opisthokonts. Such elements were likely present in some lineages early in animal mitochondrial genome evolution but were subsequently lost during the reduction of intergenic regions, which occurred in the Eumetazoa lineage after the split of Porifera. Porifera acquired their elements in several independent events. Patterns of their intra-genomic dispersal can be seen in the mt genome of Vaceletia sp. PMID- 20003197 TI - Detailed molecular and clinical investigation of a child with a partial deletion of chromosome 11 (Jacobsen syndrome). AB - BACKGROUND: Jacobsen syndrome (JBS) is a rare chromosomal disorder leading to multiple physical and mental impairment. This syndrome is caused by a partial deletion of chromosome 11, especially subband 11q24.1 has been proven to be involved. Clinical cases may easily escape diagnosis, however pancytopenia or thrombocytopenia may be indicative for JBS. RESULTS: We report a 7.5 years old boy presenting with speech development delay, hearing impairment and abnormal platelet function. High resolution SNP oligonucleotide microarray analysis revealed a terminal deletion of 11.4 Mb in size, in the area 11q24.1-11qter. This specific deletion encompasses around 170 genes. Other molecular techniques such as fluorescence in situ hybridization and multiplex ligation-dependent probe amplification were used to confirm the array-result. DISCUSSION: Our results suggest that the identification and detailed analysis of similar patients with abnormal platelet function and otherwise mild clinical features will contribute to identification of more patients with 11q deletion and JBS. PMID- 20003198 TI - Transforming growth factor-beta inhibits aromatase gene transcription in human trophoblast cells via the Smad2 signaling pathway. AB - BACKGROUND: Transforming growth factor-beta (TGF-beta) is known to exert multiple regulatory functions in the human placenta, including inhibition of estrodial production. We have previously reported that TGF-beta1 decreased aromatase mRNA levels in human trophoblast cells. The objective of this study was to investigate the molecular mechanisms underlying the regulatory effect of TGF-beta1 on aromatase expression. METHODS: To determine if TGF-beta regulates aromatase gene transcription, several reporter constructs containing different lengths of the placental specific promoter of the human aromatase gene were generated. JEG-3 cells were transiently transfected with a promoter construct and treated with or without TGF-beta1. The promoter activity was measured by luciferase assays. To examine the downstream signaling molecule mediating the effect of TGF-beta on aromatase transcription, cells were transiently transfected with dominant negative mutants of TGF-beta type II (TbetaRII) and type I receptor (ALK5) receptors before TGF-beta treatment. Smad2 activation was assessed by measuring phophorylated Smad2 protein levels in cytosolic and nuclear fractions. Smad2 expression was silenced using a siRNA expression construct. Finally, aromatase mRNA half-life was determined by treating cells with actinomycin D together with TGF-beta1 and measuring aromatase mRNA levels at various time points after treatment. RESULTS AND DISCUSSION: TGF-beta1 inhibited the aromatase promoter activity in a time- and dose-dependent manner. Deletion analysis suggests that the TGF-beta1 response element resides between -422 and -117 nucleotides upstream from the transcription start site where a Smad binding element was found. The inhibitory effect of TGF-beta1 was blocked by dominant negative mutants of TbetaRII and ALK5. TGF-beta1 treatment induced Smad2 phosphorylation and translocation into the nucleus. On the other hand, knockdown of Smad2 expression reversed the inhibitory effect of TGF-beta1 on aroamtase transcription. Furthermore, TGF-beta1 accelerated the degradation of aromatase mRNA. CONCLUSION: Our results demonstrate that TGF-beta1 exerts regulatory effects on aromatase gene at both transcriptional and post-transcriptional levels. The transcriptional regulation of aromatase gene by TGF-beta1 is mediated by the canonical TGF-beta pathway involving TbetaRII, ALK5 and Smad2. These findings further support the role of TGF-beta1 in regulating human placental functions and pregnancy. PMID- 20003199 TI - Multiplex real-time quantitative PCR, microscopy and rapid diagnostic immuno chromatographic tests for the detection of Plasmodium spp: performance, limit of detection analysis and quality assurance. AB - BACKGROUND: Accurate laboratory diagnosis of malaria species in returning travelers is paramount in the treatment of this potentially fatal infectious disease. MATERIALS AND METHODS: A total of 466 blood specimens from returning travelers to Africa, Asia, and South/Central America with suspected malaria infection were collected between 2007 and 2009 at the reference public health laboratory. These specimens were assessed by reference microscopy, multipex real time quantitative polymerase chain reaction (QPCR), and two rapid diagnostic immuno-chromatographic tests (ICT) in a blinded manner. Key clinical laboratory parameters such as limit of detection (LOD) analysis on clinical specimens by parasite stage, inter-reader variability of ICTs, staffing implications, quality assurance and cost analysis were evaluated. RESULTS: QPCR is the most analytically sensitive method (sensitivity 99.41%), followed by CARESTART (sensitivity 88.24%), and BINAXNOW (sensitivity 86.47%) for the diagnosis of malaria in returning travelers when compared to reference microscopy. However, microscopy was unable to specifically identify Plasmodia spp. in 18 out of 170 positive samples by QPCR. Moreover, the 17 samples that were negative by microscopy and positive by QPCR were also positive by ICTs. Quality assurance was achieved for QPCR by exchanging a blinded proficiency panel with another reference laboratory. The Kappa value of inter-reader variability among three readers for BINAXNOW and CARESTART was calculated to be 0.872 and 0.898 respectively. Serial dilution studies demonstrated that the QPCR cycle threshold correlates linearly with parasitemia (R(2) = 0.9746) in a clinically relevant dynamic range and retains a LOD of 11 rDNA copies/microl for P. falciparum, which was several log lower than reference microscopy and ICTs. LOD for QPCR is affected not only by parasitemia but the parasite stage distribution of each clinical specimen. QPCR was approximately 6-fold more costly than reference microscopy. DISCUSSION: These data suggest that multiplex QPCR although more costly confers a significant diagnostic advantage in terms of LOD compared to reference microscopy and ICTs for all four species. Quality assurance of QPCR is essential to the maintenance of proficiency in the clinical laboratory. ICTs showed good concordance between readers however lacked sensitivity for non falciparum species due to antigenic differences and low parasitemia. CONCLUSION: Multiplex QPCR but not ICTs is an essential adjunct to microscopy in the reference laboratory detection of malaria species specifically due to the superior LOD. ICTs are better suited to the non-reference laboratory where lower specimen volumes challenge microscopy proficiency in the non-endemic setting. PMID- 20003200 TI - Intensive insulin treatment improves forearm blood flow in critically ill patients: a randomized parallel design clinical trial. AB - INTRODUCTION: Intensive insulin treatment of critically ill patients was seen as a promising method of treatment, though recent studies showed that reducing the blood glucose level below 6 mmol/l had a detrimental outcome. The mechanisms of the effects of insulin in the critically ill are not completely understood. The purpose of the study was to test the hypothesis that intensive insulin treatment may influence forearm blood flow independently of global hemodynamic indicators. METHODS: The study encompassed 29 patients of both sexes who were admitted to the intensive care unit due to sepsis and required artificial ventilation as the result of acute respiratory failure. 14 patients were randomly selected for intensive insulin treatment (Group 1; blood glucose concentration 4.4-6.1 mmol/l), and 15 were selected for conventional insulin treatment (Group 2; blood glucose level 7.0 mmol/l-11.0 mmol/l). At the start of the study (t0, beginning up to 48 hours after admittance and the commencement of artificial ventilation), at 2 hours (t1), 24 hours (t2), and 72 hours (t3) flow in the forearm was measured for 60 minutes using the strain-gauge plethysmography method. Student's t-test of independent samples was used for comparisons between the two groups, and Mann-Whitney's U-test where appropriate. Linear regression analysis and the Pearson correlation coefficient were used to determine the levels of correlation. RESULTS: The difference in 60-minute forearm flow at the start of the study (t0) was not statistically significant between groups, while at t2 and t3 significantly higher values were recorded in Group 1 (t2; Group 1: 420.6 +/- 188.8 ml/100 ml tissue; Group 2: 266.1 +/- 122.2 ml/100 ml tissue (95% CI 30.9 278.0, P = 0.02); t3; Group 1: 369.9 +/- 150.3 ml/100 ml tissue; Group 2: 272.6 +/- 85.7 ml/100 ml tissue (95% CI 5.4-190.0, P = 0.04). At t1 a trend towards significantly higher values in Group 1 was noted (P = 0.05). The level of forearm flow was related to the amount of insulin infusion (r = 0.40). CONCLUSIONS: Compared to standard treatment, intensive insulin treatment of critically ill patients increases forearm flow. Flow increase was weakly related to the insulin dose, though not to blood glucose concentration. TRIAL REGISTRATION: TRIAL NUMBER: ISRCTN39026810. PMID- 20003201 TI - Mass action models versus the Hill model: an analysis of tetrameric human thymidine kinase 1 positive cooperativity. AB - BACKGROUND: The Hill coefficient characterizes the extent to which an enzyme exhibits positive or negative cooperativity, but it provides no information regarding the mechanism of cooperativity. In contrast, models based on the equilibrium concept of mass action can suggest mechanisms of cooperativity, but there are often many such models and often many with too many parameters. RESULTS: Mass action models of tetrameric human thymidine kinase 1 (TK1) activity data were formed as pairs of plausible hypotheses that per site activities and binary dissociation constants are equal within contiguous stretches of the number of substrates bound. Of these, six 3-parameter models were fitted to 5 different datasets. Akaike's Information Criterion was then used to form model probability weighted averages. The literature average of the 5 model averages was K = (0.85, 0.69, 0.65, 0.51) microM and k = (3.3, 3.9, 4.1, 4.1) sec-1 where K and k are per site binary dissociation constants and activities indexed by the number of substrates bound to the tetrameric enzyme. CONCLUSION: The TK1 model presented supports both K and k positive cooperativity. Three-parameter mass action models can and should replace the 3-parameter Hill model. PMID- 20003202 TI - Gastric cancer surgery: Billroth I or Billroth II for distal gastrectomy? AB - BACKGROUND: The selection of an anastomosis method after a distal gastrectomy is a highly debatable topic; however, the available documentation lacks the necessary research based on a comparison of early postoperative complications. This study was conducted to investigate the difference of early postoperative complications between Billroth I and Billroth II types of anastomosis for distal gastrectomies. METHODS: A total of 809 patients who underwent distal gastrectomies for gastric cancer during four years were included in the study. The only study endpoint was analysis of in-patients' postoperative complications. The risk adjusted complication rate was compared by POSSUM (Physiological and operative severity score for enumeration of morbidity and mortality) and the severity of complications was compared by Rui Jin Hospital classification of complication. RESULTS: Complication rate of Billroth II type of anastomosis was almost double of that in Billroth I (P=0.000). Similarly, the risk adjusted complication rate was also higher in Billroth II group. More severe complications were observed and the postoperative duration was significantly longer in Billroth II type (P=0.000). Overall expenditure was significantly higher in Billroth II type (P=0.000). CONCLUSION: Billroth II method of anastomosis was associated with higher rate of early postoperative complications. Therefore, we conclude that the Billroth I method should be the first choice after a distal gastrectomy as long as the anatomic and oncological environment of an individual patient allows us to perform it. However more prospective studies should be designed to compare the overall surgical outcomes of both anastomosis methods. PMID- 20003203 TI - Automated mass action model space generation and analysis methods for two reactant combinatorially complex equilibriums: an analysis of ATP-induced ribonucleotide reductase R1 hexamerization data. AB - BACKGROUND: Ribonucleotide reductase is the main control point of dNTP production. It has two subunits, R1, and R2 or p53R2. R1 has 5 possible catalytic site states (empty or filled with 1 of 4 NDPs), 5 possible s-site states (empty or filled with ATP, dATP, dTTP or dGTP), 3 possible a-site states (empty or filled with ATP or dATP), perhaps two possible h-site states (empty or filled with ATP), and all of this is folded into an R1 monomer-dimer-tetramer-hexamer equilibrium where R1 j-mers can be bound by variable numbers of R2 or p53R2 dimers. Trillions of RNR complexes are possible as a result. The problem is to determine which are needed in models to explain available data. This problem is intractable for 10 reactants, but it can be solved for 2 and is here for R1 and ATP. RESULTS: Thousands of ATP-induced R1 hexamerization models with up to three (s, a and h) ATP binding sites per R1 subunit were automatically generated via hypotheses that complete dissociation constants are infinite and/or that binary dissociation constants are equal. To limit the model space size, it was assumed that s-sites are always filled in oligomers and never filled in monomers, and to interpret model terms it was assumed that a-sites fill before h-sites. The models were fitted to published dynamic light scattering data. As the lowest Akaike Information Criterion (AIC) of the 3-parameter models was greater than the lowest of the 2-parameter models, only models with up to 3 parameters were fitted. Models with sums of squared errors less than twice the minimum were then partitioned into two groups: those that contained no occupied h-site terms (508 models) and those that contained at least one (1580 models). Normalized AIC densities of these two groups of models differed significantly in favor of models that did not include an h-site term (Kolmogorov-Smirnov p < 1 x 10(-15)); consistent with this, 28 of the top 30 models (ranked by AICs) did not include an h-site term and 28/30 > 508/2088 with p < 2 x 10(-15). Finally, 99 of the 2088 models did not have any terms with ATP/R1 ratios >1.5, but of the top 30, there were 14 such models (14/30 > 99/2088 with p < 3 x 10(-16)), i.e. the existence of R1 hexamers with >3 a-sites occupied by ATP is also not supported by this dataset. CONCLUSION: The analysis presented suggests that three a-sites may not be occupied by ATP in R1 hexamers under the conditions of the data analyzed. If a sites fill before h-sites, this implies that the dataset analyzed can be explained without the existence of an h-site. PMID- 20003204 TI - A microarray-based system for the simultaneous analysis of single nucleotide polymorphisms in human genes involved in the metabolism of anti-malarial drugs. AB - BACKGROUND: In order to provide a cost-effective tool to analyse pharmacogenetic markers in malaria treatment, DNA microarray technology was compared with sequencing of polymerase chain reaction (PCR) fragments to detect single nucleotide polymorphisms (SNPs) in a larger number of samples. METHODS: The microarray was developed to affordably generate SNP data of genes encoding the human cytochrome P450 enzyme family (CYP) and N-acetyltransferase-2 (NAT2) involved in anti-malarial drug metabolisms and with known polymorphisms, i.e. CYP2A6, CYP2B6, CYP2C8, CYP2C9, CYP2C19, CYP2D6, CYP3A4, CYP3A5, and NAT2. RESULTS: For some SNPs, i.e. CYP2A6*2, CYP2B6*5, CYP2C8*3, CYP2C9*3/*5, CYP2C19*3, CYP2D6*4 and NAT2*6/*7/*14, agreement between both techniques ranged from substantial to almost perfect (kappa index between 0.61 and 1.00), whilst for other SNPs a large variability from slight to substantial agreement (kappa index between 0.39 and 1.00) was found, e.g. CYP2D6*17 (2850C>T), CYP3A4*1B and CYP3A5*3. CONCLUSION: The major limit of the microarray technology for this purpose was lack of robustness and with a large number of missing data or with incorrect specificity. PMID- 20003205 TI - Comparative, double-blind, controlled study of intra-articular hyaluronic acid (Hyalubrix) injections versus local anesthetic in osteoarthritis of the hip. AB - INTRODUCTION: Comparison of intra-articular bacterial-derived hyaluronic acid (Hyalubrix) (HA) with local analgesia (mepivacaine) for osteoarthritis (OA) of the hip. METHODS: A pilot prospective, double-blind, 6-month randomized trial of 42 patients with hip OA. HA or mepivacaine was administered twice (once a month) under ultrasound guidance. Efficacy measurements included the Lequesne's algofunctional index, a visual analog scale for pain, concomitant use of analgesia, patient and physician global measurement, and safety. RESULTS: Patients in the HA group exhibited a significantly reduced Lequesne's algofunctional index 3 and 6 months after treatment (P < 0.001) and significantly reduced visual analog scale pain scores 3 and 6 months after treatment (P < 0.05) compared with the local anesthetic group. All primary and secondary measures were significantly improved versus baseline, but other than the above were not different from each other at 3 or 6 months. Adverse effects were minimal. CONCLUSIONS: This comparative study suggests a beneficial effect and safety of intra-articular HA in the management of hip OA. TRIAL REGISTRATION NUMBER: ISRCTN39397064. PMID- 20003206 TI - Identifying differentially methylated genes using mixed effect and generalized least square models. AB - BACKGROUND: DNA methylation plays an important role in the process of tumorigenesis. Identifying differentially methylated genes or CpG islands (CGIs) associated with genes between two tumor subtypes is thus an important biological question. The methylation status of all CGIs in the whole genome can be assayed with differential methylation hybridization (DMH) microarrays. However, patient samples or cell lines are heterogeneous, so their methylation pattern may be very different. In addition, neighboring probes at each CGI are correlated. How these factors affect the analysis of DMH data is unknown. RESULTS: We propose a new method for identifying differentially methylated (DM) genes by identifying the associated DM CGI(s). At each CGI, we implement four different mixed effect and generalized least square models to identify DM genes between two groups. We compare four models with a simple least square regression model to study the impact of incorporating random effects and correlations. CONCLUSIONS: We demonstrate that the inclusion (or exclusion) of random effects and the choice of correlation structures can significantly affect the results of the data analysis. We also assess the false discovery rate of different models using CGIs associated with housekeeping genes. PMID- 20003208 TI - Design and methods for a randomized clinical trial of a diabetes self-management intervention for low-income Latinos: Latinos en Control. AB - BACKGROUND: US Latinos have greater prevalence of type 2 diabetes (diabetes), uncontrolled diabetes and diabetes co-morbidities compared to non-Latino Whites. They also have lower literacy levels and are more likely to live in poverty. Interventions are needed to improve diabetes control among low-income Latinos. METHODS AND DESIGN: This randomized clinical trial tested the efficacy of a culturally- and literacy-tailored diabetes self-management intervention (Latinos en Control) on glycemic control among low-income Latinos with diabetes, compared to usual care (control). Participants were recruited from five community health centers (CHCs) in Massachusetts. The theory-based intervention included an intensive phase of 12 weekly sessions and a follow-up maintenance phase of 8 monthly sessions. Assessments occurred at baseline, and at 4 and 12 months. The primary outcome was glycosylated hemoglobin (HbA1c). Secondary outcomes were self management behaviors, weight, lipids and blood pressure. Additional outcomes included diabetes knowledge, self-efficacy, depression and quality of life. The study was designed for recruitment of 250 participants (estimated 20% dropout rate) to provide 90% power for detecting a 7% or greater change in HbA1c between the intervention and control groups. This is a difference in change of HbA1c of 0.5 to 0.6%. DISCUSSION: Low-income Latinos bear a great burden of uncontrolled diabetes and are an understudied population. Theory-based interventions that are tailored to the needs of this high-risk population have potential for improving diabetes self-management and reduce health disparities. This article describes the design and methods of a theory driven intervention aimed at addressing this need. PMID- 20003209 TI - Transcriptional profile of isoproterenol-induced cardiomyopathy and comparison to exercise-induced cardiac hypertrophy and human cardiac failure. AB - BACKGROUND: Isoproterenol-induced cardiac hypertrophy in mice has been used in a number of studies to model human cardiac disease. In this study, we compared the transcriptional response of the heart in this model to other animal models of heart failure, as well as to the transcriptional response of human hearts suffering heart failure. RESULTS: We performed microarray analyses on RNA from mice with isoproterenol-induced cardiac hypertrophy and mice with exercise induced physiological hypertrophy and identified 865 and 2,534 genes that were significantly altered in pathological and physiological cardiac hypertrophy models, respectively. We compared our results to 18 different microarray data sets (318 individual arrays) representing various other animal models and four human cardiac diseases and identified a canonical set of 64 genes that are generally altered in failing hearts. We also produced a pairwise similarity matrix to illustrate relatedness of animal models with human heart disease and identified ischemia as the human condition that most resembles isoproterenol treatment. CONCLUSION: The overall patterns of gene expression are consistent with observed structural and molecular differences between normal and maladaptive cardiac hypertrophy and support a role for the immune system (or immune cell infiltration) in the pathology of stress-induced hypertrophy. Cross-study comparisons such as the results presented here provide targets for further research of cardiac disease that might generally apply to maladaptive cardiac stresses and are also a means of identifying which animal models best recapitulate human disease at the transcriptional level. PMID- 20003210 TI - Structural issues affecting mixed methods studies in health research: a qualitative study. AB - BACKGROUND: Health researchers undertake studies which combine qualitative and quantitative methods. Little attention has been paid to the structural issues affecting this mixed methods approach. We explored the facilitators and barriers to undertaking mixed methods studies in health research. METHODS: Face-to-face semi-structured interviews with 20 researchers experienced in mixed methods research in health in the United Kingdom. RESULTS: Structural facilitators for undertaking mixed methods studies included a perception that funding bodies promoted this approach, and the multidisciplinary constituency of some university departments. Structural barriers to exploiting the potential of these studies included a lack of education and training in mixed methods research, and a lack of templates for reporting mixed methods articles in peer-reviewed journals. The 'hierarchy of evidence' relating to effectiveness studies in health care research, with the randomised controlled trial as the gold standard, appeared to pervade the health research infrastructure. Thus integration of data and findings from qualitative and quantitative components of mixed methods studies, and dissemination of integrated outputs, tended to occur through serendipity and effort, further highlighting the presence of structural constraints. Researchers are agents who may also support current structures - journal reviewers and editors, and directors of postgraduate training courses - and thus have the ability to improve the structural support for exploiting the potential of mixed methods research. CONCLUSION: The environment for health research in the UK appears to be conducive to mixed methods research but not to exploiting the potential of this approach. Structural change, as well as change in researcher behaviour, will be necessary if researchers are to fully exploit the potential of using mixed methods research. PMID- 20003211 TI - Pressure-dependent stress relaxation in acute respiratory distress syndrome and healthy lungs: an investigation based on a viscoelastic model. AB - INTRODUCTION: Limiting the energy transfer between ventilator and lung is crucial for ventilatory strategy in acute respiratory distress syndrome (ARDS). Part of the energy is transmitted to the viscoelastic tissue components where it is stored or dissipates. In mechanically ventilated patients, viscoelasticity can be investigated by analyzing pulmonary stress relaxation. While stress relaxation processes of the lung have been intensively investigated, non-linear interrelations have not been systematically analyzed, and such analyses have been limited to small volume or pressure ranges. In this study, stress relaxation of mechanically ventilated lungs was investigated, focusing on non-linear dependence on pressure. The range of inspiratory capacity was analyzed up to a plateau pressure of 45 cmH2O. METHODS: Twenty ARDS patients and eleven patients with normal lungs under mechanical ventilation were included. Rapid flow interruptions were repetitively applied using an automated super-syringe maneuver. Viscoelastic resistance, compliance and time constant were determined by multiple regression analysis using a lumped parameter model. This same viscoelastic model was used to investigate the frequency dependence of the respiratory system's impedance. RESULTS: The viscoelastic time constant was independent of pressure, and it did not differ between normal and ARDS lungs. In contrast, viscoelastic resistance increased non-linearly with pressure (normal: 8.4 (7.4-11.9) [median (lower - upper quartile)] to 35.2 (25.6-39.5) cmH2O.sec/L; ARDS: 11.9 (9.2-22.1) to 73.5 (56.8-98.7)cmH2O.sec/L), and viscoelastic compliance decreased non-linearly with pressure (normal: 130.1(116.9-151.3) to 37.4(34.7-46.3) mL/cmH2O; ARDS: 125.8(80.0-211.0) to 17.1(13.8-24.7)mL/cmH2O). The pulmonary impedance increased with pressure and decreased with respiratory frequency. CONCLUSIONS: Viscoelastic compliance and resistance are highly non-linear with respect to pressure and differ considerably between ARDS and normal lungs. None of these characteristics can be observed for the viscoelastic time constant. From our analysis of viscoelastic properties we cautiously conclude that the energy transfer from the respirator to the lung can be reduced by application of low inspiratory plateau pressures and high respiratory frequencies. This we consider to be potentially lung protective. PMID- 20003212 TI - Exploring biological network structure with clustered random networks. AB - BACKGROUND: Complex biological systems are often modeled as networks of interacting units. Networks of biochemical interactions among proteins, epidemiological contacts among hosts, and trophic interactions in ecosystems, to name a few, have provided useful insights into the dynamical processes that shape and traverse these systems. The degrees of nodes (numbers of interactions) and the extent of clustering (the tendency for a set of three nodes to be interconnected) are two of many well-studied network properties that can fundamentally shape a system. Disentangling the interdependent effects of the various network properties, however, can be difficult. Simple network models can help us quantify the structure of empirical networked systems and understand the impact of various topological properties on dynamics. RESULTS: Here we develop and implement a new Markov chain simulation algorithm to generate simple, connected random graphs that have a specified degree sequence and level of clustering, but are random in all other respects. The implementation of the algorithm (ClustRNet: Clustered Random Networks) provides the generation of random graphs optimized according to a local or global, and relative or absolute measure of clustering. We compare our algorithm to other similar methods and show that ours more successfully produces desired network characteristics.Finding appropriate null models is crucial in bioinformatics research, and is often difficult, particularly for biological networks. As we demonstrate, the networks generated by ClustRNet can serve as random controls when investigating the impacts of complex network features beyond the byproduct of degree and clustering in empirical networks. CONCLUSION: ClustRNet generates ensembles of graphs of specified edge structure and clustering. These graphs allow for systematic study of the impacts of connectivity and redundancies on network function and dynamics. This process is a key step in unraveling the functional consequences of the structural properties of empirical biological systems and uncovering the mechanisms that drive these systems. PMID- 20003213 TI - Filling the gap - COI barcode resolution in eastern Palearctic birds. AB - BACKGROUND: The Palearctic region supports relatively few avian species, yet recent molecular studies have revealed that cryptic lineages likely still persist unrecognized. A broad survey of cytochrome c oxidase I (COI) sequences, or DNA barcodes, can aid on this front by providing molecular diagnostics for species assignment. Barcodes have already been extensively surveyed in the Nearctic, which provides an interesting comparison to this region; faunal interchange between these regions has been very dynamic. We explored COI sequence divergence within and between species of Palearctic birds, including samples from Russia, Kazakhstan, and Mongolia. As of yet, there is no consensus on the best method to analyze barcode data. We used this opportunity to compare and contrast three different methods routinely employed in barcoding studies: clustering-based, distance-based, and character-based methods. RESULTS: We produced COI sequences from 1,674 specimens representing 398 Palearctic species. These were merged with published COI sequences from North American congeners, creating a final dataset of 2,523 sequences for 599 species. Ninety-six percent of the species analyzed could be accurately identified using one or a combination of the methods employed. Most species could be rapidly assigned using the cluster-based or distance-based approach alone. For a few select groups of species, the character based method offered an additional level of resolution. Of the five groups of indistinguishable species, most were pairs, save for a larger group comprising the herring gull complex. Up to 44 species exhibited deep intraspecific divergences, many of which corresponded to previously described phylogeographic patterns and endemism hotspots. CONCLUSION: COI sequence divergence within eastern Palearctic birds is largely consistent with that observed in birds from other temperate regions. Sequence variation is primarily congruent with taxonomic boundaries; deviations from this trend reveal overlooked biological patterns, and in some cases, overlooked species. More research is needed to further refine the taxonomic status of some Palearctic birds, but large genetic surveys such as this may facilitate this effort. DNA barcodes are a practical means for rapid species assignment, although efficient analytical methods will likely require a two tiered approach to differentiate closely related pairs of species. PMID- 20003214 TI - Combination antiretroviral drugs in PLGA nanoparticle for HIV-1. AB - BACKGROUND: Combination antiretroviral (AR) therapy continues to be the mainstay for HIV treatment. However, antiretroviral drug nonadherence can lead to the development of resistance and treatment failure. We have designed nanoparticles (NP) that contain three AR drugs and characterized the size, shape, and surface charge. Additionally, we investigated the in vitro release of the AR drugs from the NP using peripheral blood mononuclear cells (PBMCs). METHODS: Poly-(lactic-co glycolic acid) (PLGA) nanoparticles (NPs) containing ritonavir (RTV), lopinavir (LPV), and efavirenz (EFV) were fabricated using multiple emulsion-solvent evaporation procedure. The nanoparticles were characterized by electron microscopy and zeta potential for size, shape, and charge. The intracellular concentration of AR drugs was determined over 28 days from NPs incubated with PBMCs. Macrophages were imaged by fluorescent microscopy and flow cytometry after incubation with fluorescent NPs. Finally, macrophage cytotoxicity was determined by MTT assay. RESULTS: Nanoparticle size averaged 262 +/- 83.9 nm and zeta potential -11.4 +/- 2.4. AR loading averaged 4% (w/v). Antiretroviral drug levels were determined in PBMCs after 100 microg of NP in 75 microL PBS was added to media. Intracellular peak AR levels from NPs (day 4) were RTV 2.5 +/- 1.1; LPV 4.1 +/- 2.0; and EFV 10.6 +/- 2.7 microg and continued until day 28 (all AR >or= 0.9 microg). Free drugs (25 microg of each drug in 25 microL ethanol) added to PBMCs served as control were eliminated by 2 days. Fluorescence microscopy and flow cytometry demonstrated phagocytosis of NP into monocytes-derived macrophages (MDMs). Cellular MTT assay performed on MDMs demonstrated that NPs are not significantly cytotoxic. CONCLUSION: These results demonstrated AR NPs could be fabricated containing three antiretroviral drugs (RTV, LPV, EFV). Sustained release of AR from PLGA NP show high drug levels in PBMCs until day 28 without cytotoxicity. PMID- 20003215 TI - Adolescent body satisfaction: the role of perceived parental encouragement for physical activity. AB - BACKGROUND: Parents play an important role in the development of children's health behaviors, but less is known about the role of parental encouragement for physical activity (PA) on youth PA behavior and body image satisfaction. The purposes of this study were to: (1) longitudinally assess whether adolescent PA at age 15 mediates the effect of perceived parental encouragement for PA at age 15 for predicting adolescent body satisfaction at age 16, while controlling for body mass index (BMI), and (2) examine the extent to which adolescent sex moderates this association. METHODS: Participants were 379 boys and girls assessed at 15 and 16 years of age, who completed surveys as part of a larger longitudinal study in their health/physical education classes in a school district in Central Pennsylvania. Participants completed measures of their perception of parental encouragement for PA, PA behavior, body satisfaction, and height and weight to calculate BMI at age 15 and 16 (i.e., 10th and 11th grades). Pearson correlations were used to examine the association among the study variables and hierarchical regression analyses were used to predict body satisfaction at age 16. RESULTS: Perceived encouragement for PA from fathers, but not mothers, at age 15, was significantly associated with adolescent PA at age 15 and body satisfaction scores at age 16. Adolescents reporting higher PA behavior and perceived encouragement for PA from fathers at age 15 had higher body satisfaction scores at age 16. Moreover, adolescent PA at age 15 mediated the association between perceived fathers' encouragement for PA at age 15 and adolescent body satisfaction at age 16, when controlling for BMI. Examining the moderating effect of adolescent sex on this association revealed that adolescent PA no longer mediated the association between perceived encouragement for PA from fathers and adolescent body satisfaction, and sex moderated this association. DISCUSSION: These findings suggest that, regardless of adolescent BMI, fathers may play an instrumental role in adolescents' body image satisfaction by positively influencing PA behavior. However, the influence of perceived encouragement for PA from fathers on adolescent body satisfaction and PA behavior may differ for boys and girls. PMID- 20003217 TI - Macromolecular and elemental composition analysis and extracellular metabolite balances of Pichia pastoris growing at different oxygen levels. AB - BACKGROUND: Analysis of the cell operation at the metabolic level requires collecting data of different types and to determine their confidence level. In addition, the acquired information has to be combined in order to obtain a consistent operational view. In the case of Pichia pastoris, information of its biomass composition at macromolecular and elemental level is scarce particularly when different environmental conditions, such as oxygen availability or, genetic backgrounds (e.g. recombinant protein production vs. non production conditions) are compared. RESULTS: P. pastoris cells growing in carbon-limited chemostat cultures under different oxygenation conditions (% O2 in the bioreactor inlet gas: 21%, 11% and 8%, corresponding to normoxic, oxygen-limiting and hypoxic conditions, respectively), as well as under recombinant protein (antibody fragment, Fab) producing and non-producing conditions, were analyzed from different points of view. On the one hand, the macromolecular and elemental composition of the biomass was measured using different techniques at the different experimental conditions and proper reconciliation techniques were applied for gross error detection of the measured substrates and products conversion rates. On the other hand, fermentation data was analyzed applying elemental mass balances. This allowed detecting a previously missed by-product secreted under hypoxic conditions, identified as arabinitol (aka. arabitol). After identification of this C5 sugar alcohol as a fermentation by-product, the mass balances of the fermentation experiments were validated. CONCLUSIONS: After application of a range of analytical and statistical techniques, a consistent view of growth parameters and compositional data of P. pastoris cells growing under different oxygenation conditions was obtained. The obtained data provides a first view of the effects of oxygen limitation on the physiology of this microorganism, while recombinant Fab production seems to have little or no impact at this level of analysis. Furthermore, the results will be highly useful in other complementary quantitative studies of P. pastoris physiology, such as metabolic flux analysis. PMID- 20003218 TI - Minimal stress shielding with a Mallory-Head titanium femoral stem with proximal porous coating in total hip arthroplasty. AB - BACKGROUND: As longevity of cementless femoral components enters the third decade, concerns arise with long-term effects of fixation mode on femoral bone morphology. We examined the long-term consequences on femoral remodeling following total hip arthroplasty with a porous plasma-sprayed tapered titanium stem. METHODS: Clinical data and radiographs were reviewed from a single center for 97 randomly selected cases implanted with the Mallory-Head Porous femoral component during primary total hip arthroplasty. Measurements were taken from preoperative and long-term follow-up radiographs averaging 14 years postoperative. Average changes in the proximal, middle and diaphyseal zones were determined. RESULTS: On anteroposterior radiographs, the proximal cortical thickness was unchanged medially and the lateral zone increased 1.3%. Middle cortical thickness increased 4.3% medially and 1.2% laterally. Distal cortical thickness increased 9.6% medially and 1.9% laterally. Using the anteroposterior radiographs, canal fill at 100 mm did not correlate with bony changes at any level (Spearman's rank correlation coefficient of -0.18, 0.05, and 0.00; p value = 0.09, 0.67, 0.97). On lateral radiographs, the proximal cortical thickness increased 1.5% medially and 0.98% laterally. Middle cortical thickness increased 2.4% medially and 1.3% laterally. Distal cortical thickness increased 3.5% medially and 2.1% laterally. From lateral radiographs, canal fill at 100 mm correlated with bony hypertrophy at the proximal, mid-level, and distal femur (Spearman's rank correlation coefficient of 0.85, 0.33, and 0.28, respectively; p value = 0.001, 0.016, and 0.01, respectively). CONCLUSION: Stress shielding is minimized with the Mallory-Head titanium tapered femoral stem with circumferential proximal plasma-sprayed coating in well-fixed and well functioning total hip arthroplasty. Additionally, the majority of femora demonstrated increased cortical thickness in all zones around the stem prosthesis. LEVEL OF EVIDENCE: Therapeutic Level III. PMID- 20003216 TI - The emerging role of cardiovascular risk factor-induced mitochondrial dysfunction in atherogenesis. AB - An important role in atherogenesis is played by oxidative stress, which may be induced by common risk factors. Mitochondria are both sources and targets of reactive oxygen species, and there is growing evidence that mitochondrial dysfunction may be a relevant intermediate mechanism by which cardiovascular risk factors lead to the formation of vascular lesions. Mitochondrial DNA is probably the most sensitive cellular target of reactive oxygen species. Damage to mitochondrial DNA correlates with the extent of atherosclerosis. Several cardiovascular risk factors are demonstrated causes of mitochondrial damage. Oxidized low density lipoprotein and hyperglycemia may induce the production of reactive oxygen species in mitochondria of macrophages and endothelial cells. Conversely, reactive oxygen species may favor the development of type 2 diabetes mellitus, mainly through the induction of insulin resistance. Similarly - in addition to being a cause of endothelial dysfunction, reactive oxygen species and subsequent mitochondrial dysfunction - hypertension may develop in the presence of mitochondrial DNA mutations. Finally, other risk factors, such as aging, hyperhomocysteinemia and cigarette smoking, are also associated with mitochondrial damage and an increased production of free radicals. So far clinical studies have been unable to demonstrate that antioxidants have any effect on human atherogenesis. Mitochondrial targeted antioxidants might provide more significant results. PMID- 20003219 TI - Health care seeking among pulmonary tuberculosis suspects and patients in rural Ethiopia: a community-based study. AB - BACKGROUND: Health care seeking is a dynamic process that is influenced by socio demographic, cultural and other factors. In Ethiopia, there are limited studies regarding the health seeking behaviour of tuberculosis (TB) suspects and TB patients. However, a thorough understanding of patients' motivation and actions is crucial to understanding TB and the treatment of disease. Such insights would conceivably help to reduce delay in diagnosis, improve treatment adherence and thereby reduce transmission of TB in the community. The objective of this study was to describe and analyze health care seeking among TB suspects and pulmonary TB (PTB) cases in a rural district of the Amhara Region in Ethiopia. METHODS: Study kebeles were randomly selected in a cross-sectional study design. House-to house visits were conducted in which individuals aged 15 years and above in all households of the kebeles were included. Subjects with symptoms suggestive of TB were interviewed about their health seeking behaviour, socio-demographic and clinical factors using a semi-structured questionnaire. Logistics regression analysis was employed to assess associations between the independent and outcome variables. RESULTS: The majority, 787 (78%), TB suspects and 33 (82.5%) PTB cases had taken health care actions for symptoms from sources outside their homes. The median delay before the first action was 30 days. In logistics regression, women (AOR 0.8, 95% CI 0.6, 0.9) were found to be less likely to visit a medical health provider than men. Those with a long duration of cough (AOR 1.5, 95% CI 1.03, 2.1) and those with a previous history of TB (AOR 1.5, 95% CI 1.03, 2.3) were more likely to visit a medical health provider compared to those with a shorter duration of cough and with no history of TB. CONCLUSION: The majority of TB suspects and PTB cases had already taken health care actions for their symptoms at the time of the survey. The availability of a simple and rapid diagnostic TB test for use at the lowest level of health care and the involvement of all health providers in case finding activities are imperative for early TB case detection. PMID- 20003220 TI - RFX2 is a candidate downstream amplifier of A-MYB regulation in mouse spermatogenesis. AB - BACKGROUND: Mammalian spermatogenesis involves formation of haploid cells from the male germline and then a complex morphological transformation to generate motile sperm. Focusing on meiotic prophase, some tissue-specific transcription factors are known (A-MYB) or suspected (RFX2) to play important roles in modulating gene expression in pachytene spermatocytes. The current work was initiated to identify both downstream and upstream regulatory connections for Rfx2. RESULTS: Searches of pachytene up-regulated genes identified high affinity RFX binding sites (X boxes) in promoter regions of several new genes: Adam5, Pdcl2, and Spag6. We confirmed a strong promoter-region X-box for Alf, a germ cell-specific variant of general transcription factor TFIIA. Using Alf as an example of a target gene, we showed that its promoter is stimulated by RFX2 in transfected cells and used ChIP analysis to show that the promoter is occupied by RFX2 in vivo. Turning to upstream regulation of the Rfx2 promoter, we identified a cluster of three binding sites (MBS) for the MYB family of transcription factors. Because testis is one of the few sites of A-myb expression, and because spermatogenesis arrests in pachytene in A-myb knockout mice, the MBS cluster implicates Rfx2 as an A-myb target. Electrophoretic gel-shift, ChIP, and co transfection assays all support a role for these MYB sites in Rfx2 expression. Further, Rfx2 expression was virtually eliminated in A-myb knockout testes. Immunohistology on testis sections showed that A-MYB expression is up-regulated only after pachytene spermatocytes have clearly moved away from the tubule wall, which correlates with onset of RFX2 expression, whereas B-MYB expression, by contrast, is prevalent only in earlier spermatocytes and spermatogonia. CONCLUSION: With an expanding list of likely target genes, RFX2 is potentially an important transcriptional regulator in pachytene spermatocytes. Rfx2 itself is a good candidate to be regulated by A-MYB, which is essential for meiotic progression. If Alf is a genuine RFX2 target, then A-myb, Rfx2, and Alf may form part of a transcriptional network that is vital for completion of meiosis and preparation for post-meiotic differentiation. PMID- 20003221 TI - Direct rosiglitazone action on steroidogenesis and proinflammatory factor production in human granulosa-lutein cells. AB - BACKGROUND: Ovarian granulosa cells are the predominant source of estradiol and progesterone biosynthesis in vivo. Rosiglitazone, a synthetic agonist of the peroxisome proliferator-activated receptor gamma (PPAR gamma), is applied as the treatment of insulin resistance including women with PCOS. The aim of the study was to investigate the direct effects of rosiglitazone on steroidogenesis and proinflammatory factor production in human granulosa-lutein cells (GLCs). METHODS: Primary human GLCs were separated during in vitro fertilization and cultured in the presence of rosiglitazone, GW9662 (an antagonist of PPAR gamma) and hCG. The mRNA expression of key steroidogenic factors including 3beta- hydroxysteriod dehydrogenase (3beta-HSD), cytochrome P-450 scc (CYP11A1), cytochrome P-450 aromatase (CYP19A1), and steroidogenic acute regulatory protein (StAR) were detected by quantitative real-time PCR. Estradiol and progesterone levels in GLCs cultures were measured by chemiluminescence immunoassay, and the proinflammtory factors (TNFalpha and IL-6) in conditioned culture media were measured by ELISA. RESULTS: PPAR gamma mRNA levels increased up to 3.24 fold by rosiglitazone at the concentration of 30 microM compared to control (P<0.05). hCG alone or hCG with rosiglitazone had no significant effects on PPAR gamma mRNA levels. The CYP19A1 mRNA level at exposure to rosiglitazone alone showed a drop, but was not significantly reduced comparing to control. The expression levels of enzymes 3beta-HSD and CYP11A1 in all treatments did not alter significantly. The StAR mRNA expression at exposure to rosiglitazone was significantly increased comparing to control (P<0.05). The media concentrations of E2 and progesterone by rosiglitazone treatment showed a declining trend comparing to control or cotreatment with hCG, which did not reach significance. Most importantly, treatment with rosiglitazone decreased TNFalpha secretion in a statistically significant manner compared with control (P<0.05). The concentration of IL-6 following rosiglitazone exposure did not significantly decrease comparing to control. CONCLUSION: In cultured GLCs, rosiglitazone stimulated StAR expression, but did not significantly affect steroidogenic enzymes, as well as E2 and progesterone production. Moreover, rosiglitazone significantly decreased the production of TNFalpha in human GLCs, suggesting that PPAR gamma may play a role in the regulation of GLCs functions through inhibiting proinflammatory factors. PMID- 20003222 TI - Fatal hemopericardial tamponade due to primary pericardial mesothelioma: a case report. AB - BACKGROUND: Primary mesothelioma of the pericardium comprises less than 1% of all mesothelioma cases. Its typical presentation is insidious, with nonspecific signs and symptoms, and usually results in heart failure due to cardiac tamponade, either by a serous effusion or by direct tumorous constriction of the heart. With the exception of several case reports, the outcome is uniformly fatal, and patients typically die within six months of diagnosis. CASE PRESENTATION: A 45 year-old African American female presented to the emergency department with several days of dizziness, difficulty walking, and low blood pressure. The patient suddenly suffered cardiac arrest, and her death was pronounced. The medical examiner assumed jurisdiction of the case due to the sudden death nature of the case without known medical history. At autopsy, a one-liter hemopericardium was present, and the pericardial sac was thick, granular and adhesed to the heart, suspicious for pericarditis. Microscopic examination of the pericardial tissue instead led to a diagnosis of primary pericardial mesothelioma. CONCLUSION: Our case demonstrates a pericardial mesothelioma initially masquerading grossly as pericarditis. Microscopic examination of any grossly abnormal pericardial tissue therefore may be warranted so that a neoplastic disease process does not go undetected. Additionally, of the approximately 200 such tumors reported in the medical literature, a case demonstrating marked hemopericardium and resulting in sudden death has not been described until now. PMID- 20003223 TI - Matrix metalloproteinase 28, a novel matrix metalloproteinase, is constitutively expressed in human intervertebral disc tissue and is present in matrix of more degenerated discs. AB - INTRODUCTION: The regulation and elevation in expression of the catabolic matrix metalloproteinases (MMPs) is of high importance in the human intervertebral disc since upregulation of these matrix-degrading enzymes results in matrix destruction associated with disc degeneration. MMP28 (epilysin) is a newly discovered MMP believed to play a role in matrix composition and turnover in skin. It is present in basal keratinocytes where its expression is upregulated with wound repair, and in cartilage and synovium where it is upregulated in osteoarthritis. Recent work has shown that mechanical compression can act to modulate expression of MMP28. The expression of MMP28 is unexplored in the intervertebral disc. METHODS: Following approval by our human subjects institutional review board, we employed microarray analyses to evaluate in vivo expression of MMP28 and the MMP28 precursor in human disc tissue, and utilized immunohistochemistry to determine cellular and extracellular matrix localization of MMP28 in 35 human disc tissue specimens. The percentage of cells positive for MMP28 immunocytochemical localization was also determined. RESULTS: The present work documents the expression and presence of MMP28 in cells and extracellular matrix (ECM) of the human intervertebral disc. Gene expression levels in human disc tissue were detectable for both MMP28 and the MMP28 precursor. MMP28 cytoplasmic localization was present in cells of the outer annulus; it was also present in some, but not all, cells of the inner annulus and nucleus. MMP28 was not found in the ECM of healthier Grade I to II discs, but was identified in the ECM of 61% of the more degenerated Grade III to V discs (P = 0.0018). There was a significant difference in cellular MMP28 distribution in the disc (P = 0.008): the outer annulus showed the largest percentage of cells positive for MMP28 immunolocalization, followed by the inner annulus and then the nucleus. Herniated discs showed a significantly greater proportion of MMP28-positive cells compared with nonherniated discs (P = 0.034). CONCLUSIONS: Findings presented here show the first documentation of intervertebral disc expression and production of MMP28. MMP28 was found in both disc cell cytoplasm and in the ECM of more degenerated specimens, with greater cellular localization in the outer annulus and in herniated disc specimens. These findings are important because of the key role of MMPs in disc turnover and homeostasis, and previous indications of a role for this MMP in matrix repair and matrix turnover in other tissues. Our data, which show the presence of MMP28 in human disc tissue, suggest that MMP28 may have a potentially important role in ECM modulation in the healthy and degenerating disc. PMID- 20003224 TI - Aerobic exercise in obese diabetic patients with chronic kidney disease: a randomized and controlled pilot study. AB - BACKGROUND: Patients with obesity, diabetes, and chronic kidney disease (CKD) are generally physically inactive, have a high mortality rate, and may benefit from an exercise program. METHODS: We performed a 24-week randomized controlled feasibility study comparing aerobic exercise plus optimal medical management to medical management alone in patients with type 2 diabetes, obesity (body mass index [BMI] > 30 kg/m2), and stage 2-4 CKD (estimated glomerular filtration rate [eGFR] 15-90 mL/min/1.73 m2 with persistent proteinuria). Subjects randomized to exercise underwent thrice weekly aerobic training for 6 followed by 18 weeks of supervised home exercise. The primary outcome variable was change in proteinuria. RESULTS: Seven subjects randomized to exercise and 4 control subjects completed the study. Exercise training resulted in an increase in exercise duration during treadmill testing, which was accompanied by slight but insignificant decreases in resting systolic blood pressure and 24-hour proteinuria. Exercise did not alter GFR, hemoglobin, glycated hemoglobin, serum lipids, or C-reactive protein (CRP). Caloric intake and body weight and composition also did not change with exercise training. CONCLUSION: Exercise training in obese diabetic patients with CKD is feasible and may have clinical benefits. A large-scale randomized controlled trial to determine the effects of exercise on renal functions, cardiovascular fitness, inflammation, and oxidative stress in diabetic patients with CKD is planned. PMID- 20003225 TI - An experimental assessment of in silico haplotype association mapping in laboratory mice. AB - BACKGROUND: To assess the utility of haplotype association mapping (HAM) as a quantitative trait locus (QTL) discovery tool, we conducted HAM analyses for red blood cell count (RBC) and high density lipoprotein cholesterol (HDL) in mice. We then experimentally tested each HAM QTL using published crosses or new F2 intercrosses guided by the haplotype at the HAM peaks. RESULTS: The HAM for RBC, using 33 classic inbred lines, revealed 8 QTLs; 2 of these were true positives as shown by published crosses. A HAM-guided (C57BL/6J x CBA/J)F2 intercross we carried out verified 2 more as true positives and 4 as false positives. The HAM for HDL, using 81 strains including recombinant inbred lines and chromosome substitution strains, detected 46 QTLs. Of these, 36 were true positives as shown by published crosses. A HAM-guided (C57BL/6J x A/J)F2 intercross that we carried out verified 2 more as true positives and 8 as false positives. By testing each HAM QTL for RBC and HDL, we demonstrated that 78% of the 54 HAM peaks were true positives and 22% were false positives. Interestingly, all false positives were in significant allelic association with one or more real QTL. CONCLUSION: Because type I errors (false positives) can be detected experimentally, we conclude that HAM is useful for QTL detection and narrowing. We advocate the powerful and economical combined approach demonstrated here: the use of HAM for QTL discovery, followed by mitigation of the false positive problem by testing the HAM-predicted QTLs with small HAM-guided experimental crosses. PMID- 20003226 TI - Development and tracking of central patterns of subcutaneous fat of rural South African youth: Ellisras longitudinal study. AB - BACKGROUND: Individuals grow and accumulate central patterns of body fat into the diseases they will suffer from as older adults. The need to elicit the development and tracking of central patterns of body fat from younger age into adolescent remains to be explored. METHOD: Skinfolds measurements were done according to the standard procedures in the Ellisras Longitudinal Growth and Health Study. In total, 2,225 children--550 preschool and 1,675 primary school- aged 3-10 years (birth cohorts 1993 to 1986) were enrolled at baseline in 1996 and followed through out the eight-year periodic surveys. In 2003, 1,771 children -489 preschool and 1,282 primary school--were still in the study. RESULTS: The development of triceps, biceps, suprailiac and suscapular skinfolds of Ellisras girls were significantly higher (p < 0.001 to 0.05) compared to boys over time. The tracking coefficient between the initial measurements and the subsequent measurements was higher for skinfolds (r about 0.63) than for skinfold ratios (r about 0.43). Longitudinal tracking coefficient measuring the association between the initial measurements and all the follow up measurements simultaneously was about 0.57. CONCLUSION: The accumulation of central patterns of body fat of Ellisras children starts in childhood and adolescence spurt with Ellisras girls acquiring more than boys over time. High significant tracking of skinfold thickness while the skinfold ratios show low and insignificant tracking over time. The magnitude of central patterns of body fat accumulation over time requires further investigation to clarify their association with risk factors for cardiovascular diseases. PMID- 20003227 TI - Disruption of occludin function in polarized epithelial cells activates the extrinsic pathway of apoptosis leading to cell extrusion without loss of transepithelial resistance. AB - BACKGROUND: Occludin is a tetraspanin protein normally localized to tight junctions. The protein interacts with a variety of pathogens including viruses and bacteria, an interaction that sometimes leads to its extrajunctional localization. RESULTS: Here we report that treatment of mammary epithelial monolayers with a circularized peptide containing a four amino acid sequence found in the second extracellular loop of occludin, LHYH, leads to the appearance of extrajunctional occludin and activation of the extrinsic apoptotic pathway. At early times after peptide treatment endogenous occludin and the LYHY peptide were co-localized in extrajunctional patches, which were also shown to contain components of the death inducing signaling complex (DISC), caspases 8 and 3, the death receptor FAS and the adaptor molecule FADD. After this treatment occludin could be immunoprecipitated with FADD, confirming its interaction with the DISC. Extrusion after LYHY treatment was accomplished with no loss of epithelial resistance. CONCLUSION: These observations provide strong evidence that, following disruption, occludin forms a complex with the extrinsic death receptor leading to extrusion of apoptotic cells from the epithelial monolayer. They suggest that occludin has a protective as well as a barrier forming role in epithelia; pathogenic agents which utilize this protein as an entry point into the cell might set off an apoptotic reaction allowing extrusion of the infected cell before the pathogen can gain entry to the interstitial space. PMID- 20003228 TI - Acute perimyocarditis mimicking transmural myocardial infarction. AB - Although acute pericarditis has charachteristic electrocardiographic (ECG) findings that differentiate it from acute ST segment elevation myocardial infarction (MI); in certain cases diagnosis is somewhat difficult especially when the ECG reveals focal instead of diffuse changes and moreover when pericarditis is associated with an underlying myocarditis causing elevation of the cardiac biomarkers therefore increasing the difficulty in differentiating between both enteties. This is especially important because adverse lethal side effect can occur if thrombolytic therapy is administered for a patient with acute pericarditis, or if a diagnosis of transmural MI is missed. In this case report we are describing an 18 year old male patient who presented with an acute onset of severe chest pain associated with focal ECG changes and elevated cardiac enzymes mimicking transmural MI. This report aims to sensitize readers to this debate and create awareness among cardiologists and intensivists with both presentations and how to reach an accurate diagnosis. PMID- 20003231 TI - Embolization of cyanoacrylate glue in systemic circulation in a case of hepatocellular carcinoma: an autopsy report. AB - We report a case of embolism of the sclerosant dye with subsequent formation of foreign-body giant cell reaction within the veins of pulmonary and portal circulation in an autopsy case of hepatocellular carcinoma developing over an underlying primary biliary cirrhosis. PMID- 20003229 TI - Stratification of the severity of critically ill patients with classification trees. AB - BACKGROUND: Development of three classification trees (CT) based on the CART (Classification and Regression Trees), CHAID (Chi-Square Automatic Interaction Detection) and C4.5 methodologies for the calculation of probability of hospital mortality; the comparison of the results with the APACHE II, SAPS II and MPM II 24 scores, and with a model based on multiple logistic regression (LR). METHODS: Retrospective study of 2864 patients. Random partition (70:30) into a Development Set (DS) n = 1808 and Validation Set (VS) n = 808. Their properties of discrimination are compared with the ROC curve (AUC CI 95%), Percent of correct classification (PCC CI 95%); and the calibration with the Calibration Curve and the Standardized Mortality Ratio (SMR CI 95%). RESULTS: CTs are produced with a different selection of variables and decision rules: CART (5 variables and 8 decision rules), CHAID (7 variables and 15 rules) and C4.5 (6 variables and 10 rules). The common variables were: inotropic therapy, Glasgow, age, (A-a)O2 gradient and antecedent of chronic illness. In VS: all the models achieved acceptable discrimination with AUC above 0.7. CT: CART (0.75(0.71-0.81)), CHAID (0.76(0.72-0.79)) and C4.5 (0.76(0.73-0.80)). PCC: CART (72(69-75)), CHAID (72(69 75)) and C4.5 (76(73-79)). Calibration (SMR) better in the CT: CART (1.04(0.95 1.31)), CHAID (1.06(0.97-1.15) and C4.5 (1.08(0.98-1.16)). CONCLUSION: With different methodologies of CTs, trees are generated with different selection of variables and decision rules. The CTs are easy to interpret, and they stratify the risk of hospital mortality. The CTs should be taken into account for the classification of the prognosis of critically ill patients. PMID- 20003232 TI - The common FTO variant rs9939609 is not associated with BMI in a longitudinal study on a cohort of Swedish men born 1920-1924. AB - BACKGROUND: Common FTO (fat mass and obesity associated) gene variants have recently been strongly associated with body mass index and obesity in several large studies. Here we set out to examine the association of the FTO variant rs9939609 with BMI in a 32 year follow up study of men born 1920-1924. Moreover, we analyzed the effect of physical activity on the different genotypes. METHODS: The FTO rs9936609 was genotyped using an Illumina golden gate assay. BMI was calculated using standard methods and body fat was estimated by measuring skinfold thickness using a Harpenden caliper. Physical activity was assessed using a four question medical questionnaire. RESULTS: FTO rs9939609 was genotyped in 1153 elderly Swedish men taking part of a population-based cohort study, the ULSAM cohort. The risk of obesity and differences in BMI according to genotype at the ages of 50, 60, 70, 77 and 82 were investigated. We found no increased risk of obesity and no association with BMI at any age with the FTO rs9939609 variant. We found however interaction between physical activity at the age of 50 years and genotype on BMI levels (p = 0.039) and there was a clear trend towards larger BMI differences between the TT and AA carriers as well as between AT and AA carriers in the less physically active subjects. CONCLUSION: Here we found that the well established obesity risk allele for a common variant in FTO does not associate with increased BMI levels in a Swedish population of adult men which reached adulthood before the appearance of today's obesogenic enviroment. There is an interaction between physical activity and the effect of the FTO genotype on BMI levels suggesting that lack of physical activity is a requirement for an association of FTO gene variants to obesity. PMID- 20003233 TI - Aberrant expression and potency as a cancer immunotherapy target of alpha methylacyl-coenzyme A racemase in prostate cancer. AB - Alpha-methylacyl-CoA racemase (AMACR) is an enzyme playing an important role in the beta-oxidation of branched-chain fatty acids and fatty acid derivatives. High expression levels of AMACR have been described in various cancers, including prostate cancer, colorectal cancer and kidney cancer. Because of its cancer specific and frequent expression, AMACR could be an attractive target for cytotoxic T-lymphocyte (CTL)-based immunotherapy for cancer. In the present study, we examined the induction of AMACR-specific CTLs from prostate cancer patients' peripheral blood mononuclear cells (PBMCs) and determined HLA-A24 restricted CTL epitopes.RT-PCR and immunohistochemical analysis revealed that AMACR was strongly expressed in prostate cancer cell lines and tissues as compared with benign or normal prostate tissues. Four AMACR-derived peptides carrying the HLA-A24-binding motif were synthesized from the amino acid sequence of this protein and analyzed to determine their binding affinities to HLA-A24. By stimulating patient's PBMCs with the peptides, specific CTLs were successfully induced in 6 of 11 patients. The peptide-specific CTLs exerted significant cytotoxic activity against AMACR-expressing prostate cancer cells in the context of HLA-A24. Our study demonstrates that AMACR could become a target antigen for prostate cancer immunotherapy, and that the AMACR-derived peptides might be good peptide vaccine candidates for HLA-A24-positive AMACR-expressing cancer patients. PMID- 20003234 TI - Ttk69-dependent repression of lozenge prevents the ectopic development of R7 cells in the Drosophila larval eye disc. AB - BACKGROUND: During the development of the Drosophila eye, specific cell types differentiate from an initially equipotent group of uncommitted precursor cells. The lozenge (lz) gene, which is a member of the Runt family of transcriptional regulators, plays a pivotal role in mediating this process through regulating the expression of several fate-specifying transcription factors. However, the regulation of lz, and the control of lz expression levels in different cell types is not fully understood. RESULTS: Here, we show a genetic interaction between Tramtrack69 (Ttk69) a key transcriptional repressor and an inhibitor of neuronal fate specification, and lz, the master patterning gene of cells posterior to the morphogenetic furrow in the Drosophila eye disc. Loss of Ttk69 expression causes the development of ectopic R7 cells in the third instar eye disc, with these cells being dependent upon Lz for their development. Using the binary UAS Gal4 system, we show that overexpression of Ttk69 causes the loss of lz-dependent differentiating cells, and a down-regulation of Lz expression in the developing eye. The loss of lz-dependent cells can be rescued by overexpressing lz via a GMR lz transgene. We provide additional data showing that factors functioning upstream of Ttk69 in eye development regulate lz in a Ttk69-dependent manner. CONCLUSIONS: Our results lead us to conclude that Ttk69 can either directly or indirectly repress lz gene expression to prevent the premature development of R7 precursor cells in the developing eye of Drosophila. We therefore define a mechanism for the tight regulatory control of the master pre-patterning gene, lz, in early Drosophila eye development and provide insight into how differential levels of lz expression can be achieved to effect specific cell fate outcomes. PMID- 20003235 TI - Protein-protein docking using region-based 3D Zernike descriptors. AB - BACKGROUND: Protein-protein interactions are a pivotal component of many biological processes and mediate a variety of functions. Knowing the tertiary structure of a protein complex is therefore essential for understanding the interaction mechanism. However, experimental techniques to solve the structure of the complex are often found to be difficult. To this end, computational protein protein docking approaches can provide a useful alternative to address this issue. Prediction of docking conformations relies on methods that effectively capture shape features of the participating proteins while giving due consideration to conformational changes that may occur. RESULTS: We present a novel protein docking algorithm based on the use of 3D Zernike descriptors as regional features of molecular shape. The key motivation of using these descriptors is their invariance to transformation, in addition to a compact representation of local surface shape characteristics. Docking decoys are generated using geometric hashing, which are then ranked by a scoring function that incorporates a buried surface area and a novel geometric complementarity term based on normals associated with the 3D Zernike shape description. Our docking algorithm was tested on both bound and unbound cases in the ZDOCK benchmark 2.0 dataset. In 74% of the bound docking predictions, our method was able to find a near-native solution (interface C-alphaRMSD < or = 2.5 A) within the top 1000 ranks. For unbound docking, among the 60 complexes for which our algorithm returned at least one hit, 60% of the cases were ranked within the top 2000. Comparison with existing shape-based docking algorithms shows that our method has a better performance than the others in unbound docking while remaining competitive for bound docking cases. CONCLUSION: We show for the first time that the 3D Zernike descriptors are adept in capturing shape complementarity at the protein-protein interface and useful for protein docking prediction. Rigorous benchmark studies show that our docking approach has a superior performance compared to existing methods. PMID- 20003236 TI - Improving follow-up of abnormal cancer screens using electronic health records: trust but verify test result communication. AB - BACKGROUND: Early detection of colorectal cancer through timely follow-up of positive Fecal Occult Blood Tests (FOBTs) remains a challenge. In our previous work, we found 40% of positive FOBT results eligible for colonoscopy had no documented response by a treating clinician at two weeks despite procedures for electronic result notification. We determined if technical and/or workflow related aspects of automated communication in the electronic health record could lead to the lack of response. METHODS: Using both qualitative and quantitative methods, we evaluated positive FOBT communication in the electronic health record of a large, urban facility between May 2008 and March 2009. We identified the source of test result communication breakdown, and developed an intervention to fix the problem. Explicit medical record reviews measured timely follow-up (defined as response within 30 days of positive FOBT) pre- and post-intervention. RESULTS: Data from 11 interviews and tracking information from 490 FOBT alerts revealed that the software intended to alert primary care practitioners (PCPs) of positive FOBT results was not configured correctly and over a third of positive FOBTs were not transmitted to PCPs. Upon correction of the technical problem, lack of timely follow-up decreased immediately from 29.9% to 5.4% (p<0.01) and was sustained at month 4 following the intervention. CONCLUSION: Electronic communication of positive FOBT results should be monitored to avoid limiting colorectal cancer screening benefits. Robust quality assurance and oversight systems are needed to achieve this. Our methods may be useful for others seeking to improve follow-up of FOBTs in their systems. PMID- 20003237 TI - Identifying sexual differentiation genes that affect Drosophila life span. AB - BACKGROUND: Sexual differentiation often has significant effects on life span and aging phenotypes. For example, males and females of several species have different life spans, and genetic and environmental manipulations that affect life span often have different magnitude of effect in males versus females. Moreover, the presence of a differentiated germ-line has been shown to affect life span in several species, including Drosophila and C. elegans. METHODS: Experiments were conducted to determine how alterations in sexual differentiation gene activity might affect the life span of Drosophila melanogaster. Drosophila females heterozygous for the tudor[1] mutation produce normal offspring, while their homozygous sisters produce offspring that lack a germ line. To identify additional sexual differentiation genes that might affect life span, the conditional transgenic system Geneswitch was employed, whereby feeding adult flies or developing larvae the drug RU486 causes the over-expression of selected UAS-transgenes. RESULTS: In this study germ-line ablation caused by the maternal tudor[1] mutation was examined in a long-lived genetic background, and was found to increase life span in males but not in females, consistent with previous reports. Fitting the data to a Gompertz-Makeham model indicated that the maternal tudor[1] mutation increases the life span of male progeny by decreasing age independent mortality. The Geneswitch system was used to screen through several UAS-type and EP-type P element mutations in genes that regulate sexual differentiation, to determine if additional sex-specific effects on life span would be obtained. Conditional over-expression of transformer female isoform (traF) during development produced male adults with inhibited sexual differentiation, however this caused no significant change in life span. Over expression of doublesex female isoform (dsxF) during development was lethal to males, and produced a limited number of female escapers, whereas over-expression of dsxF specifically in adults greatly reduced both male and female life span. Similarly, over-expression of fruitless male isoform A (fru-MA) during development was lethal to both males and females, whereas over-expression of fru MA in adults greatly reduced both male and female life span. CONCLUSION: Manipulation of sexual differentiation gene expression specifically in the adult, after morphological sexual differentiation is complete, was still able to affect life span. In addition, by manipulating gene expression during development, it was possible to significantly alter morphological sexual differentiation without a significant effect on adult life span. The data demonstrate that manipulation of sexual differentiation pathway genes either during development or in adults can affect adult life span. PMID- 20003238 TI - Reanalysis of cancer mortality in Japanese A-bomb survivors exposed to low doses of radiation: bootstrap and simulation methods. AB - BACKGROUND: The International Commission on Radiological Protection (ICRP) recommended annual occupational dose limit is 20 mSv. Cancer mortality in Japanese A-bomb survivors exposed to less than 20 mSv external radiation in 1945 was analysed previously, using a latency model with non-linear dose response. Questions were raised regarding statistical inference with this model. METHODS: Cancers with over 100 deaths in the 0 - 20 mSv subcohort of the 1950-1990 Life Span Study are analysed with Poisson regression models incorporating latency, allowing linear and non-linear dose response. Bootstrap percentile and Bias corrected accelerated (BCa) methods and simulation of the Likelihood Ratio Test lead to Confidence Intervals for Excess Relative Risk (ERR) and tests against the linear model. RESULTS: The linear model shows significant large, positive values of ERR for liver and urinary cancers at latencies from 37 - 43 years. Dose response below 20 mSv is strongly non-linear at the optimal latencies for the stomach (11.89 years), liver (36.9), lung (13.6), leukaemia (23.66), and pancreas (11.86) and across broad latency ranges. Confidence Intervals for ERR are comparable using Bootstrap and Likelihood Ratio Test methods and BCa 95% Confidence Intervals are strictly positive across latency ranges for all 5 cancers. Similar risk estimates for 10 mSv (lagged dose) are obtained from the 0 20 mSv and 5 - 500 mSv data for the stomach, liver, lung and leukaemia. Dose response for the latter 3 cancers is significantly non-linear in the 5 - 500 mSv range. CONCLUSION: Liver and urinary cancer mortality risk is significantly raised using a latency model with linear dose response. A non-linear model is strongly superior for the stomach, liver, lung, pancreas and leukaemia. Bootstrap and Likelihood-based confidence intervals are broadly comparable and ERR is strictly positive by bootstrap methods for all 5 cancers. Except for the pancreas, similar estimates of latency and risk from 10 mSv are obtained from the 0 - 20 mSv and 5 - 500 mSv subcohorts. Large and significant cancer risks for Japanese survivors exposed to less than 20 mSv external radiation from the atomic bombs in 1945 cast doubt on the ICRP recommended annual occupational dose limit. PMID- 20003239 TI - Crude subcellular fractionation of cultured mammalian cell lines. AB - BACKGROUND: The expression and study of recombinant proteins in mammalian culture systems can be complicated during the cell lysis procedure by contaminating proteins from cellular compartments distinct from those within which the protein of interest resides and also by solubility issues that may arise from the use of a single lysis buffer. Partial subcellular fractionation using buffers of increasing stringency, rather than whole cell lysis is one way in which to avoid or reduce this contamination and ensure complete recovery of the target protein. Currently published protocols involve time consuming centrifugation steps which may require expensive equipment and commercially available kits can be prohibitively expensive when handling large or multiple samples. FINDINGS: We have established a protocol to sequentially extract proteins from cultured mammalian cells in fractions enriched for cytosolic, membrane bound organellar, nuclear and insoluble proteins. All of the buffers used can be made inexpensively and easily and the protocol requires no costly equipment. While the method was optimized for a specific cell type, we demonstrate that the protocol can be applied to a variety of commonly used cell lines and anticipate that it can be applied to any cell line via simple optimization of the primary extraction step. CONCLUSION: We describe a protocol for the crude subcellular fractionation of cultured mammalian cells that is both straightforward and cost effective and may facilitate the more accurate study of recombinant proteins and the generation of purer preparations of said proteins from cell extracts. PMID- 20003240 TI - In silico approaches to study mass and energy flows in microbial consortia: a syntrophic case study. AB - BACKGROUND: Three methods were developed for the application of stoichiometry based network analysis approaches including elementary mode analysis to the study of mass and energy flows in microbial communities. Each has distinct advantages and disadvantages suitable for analyzing systems with different degrees of complexity and a priori knowledge. These approaches were tested and compared using data from the thermophilic, phototrophic mat communities from Octopus and Mushroom Springs in Yellowstone National Park (USA). The models were based on three distinct microbial guilds: oxygenic phototrophs, filamentous anoxygenic phototrophs, and sulfate-reducing bacteria. Two phases, day and night, were modeled to account for differences in the sources of mass and energy and the routes available for their exchange. RESULTS: The in silico models were used to explore fundamental questions in ecology including the prediction of and explanation for measured relative abundances of primary producers in the mat, theoretical tradeoffs between overall productivity and the generation of toxic by products, and the relative robustness of various guild interactions. CONCLUSION: The three modeling approaches represent a flexible toolbox for creating cellular metabolic networks to study microbial communities on scales ranging from cells to ecosystems. A comparison of the three methods highlights considerations for selecting the one most appropriate for a given microbial system. For instance, communities represented only by metagenomic data can be modeled using the pooled method which analyzes a community's total metabolic potential without attempting to partition enzymes to different organisms. Systems with extensive a priori information on microbial guilds can be represented using the compartmentalized technique, employing distinct control volumes to separate guild-appropriate enzymes and metabolites. If the complexity of a compartmentalized network creates an unacceptable computational burden, the nested analysis approach permits greater scalability at the cost of more user intervention through multiple rounds of pathway analysis. PMID- 20003241 TI - Cadmium burden and the risk and phenotype of prostate cancer. AB - BACKGROUND: Studies on the association between prostate cancer and cadmium exposure have yielded conflicting results. This study explored cadmium burden on the risk and phenotype of prostate cancer in men with no evident environmental exposure. METHODS: Hospital-based 261 prostate cancer cases and 267 controls with benign diseases were recruited from four hospitals in Taiwan. Demographic, dietary and lifestyle data were collected by standardized questionnaires. Blood cadmium (BCd) and creatinine-adjusted urine cadmium (CAUCd) levels were measured for each participant. Statistical analyses measured the prostate cancer risk associated with BCd and CAUCd separately, controlling for age, smoking and institution. BCd and CAUCd levels within cases were compared in relation to the disease stage and the Gleason score. RESULTS: High family income, low beef intake, low dairy product consumption and positive family history were independently associated with the prostate carcinogenesis. There was no difference in BCd levels between cases and controls (median, 0.88 versus 0.87 microg/l, p = 0.45). Cases had lower CAUCd levels than controls (median, 0.94 versus 1.40 microg/g creatinine, p = 0.001). However, cases with higher BCd and CAUCd levels tended to be at more advanced stages and to have higher Gleason scores. The prostate cancer cases with Gleason scores of > or = 8 had an odds ratio of 2.89 (95% confidence interval 1.25-6.70), compared with patients with scores of 2-6. CONCLUSION: Higher CAUCd and BCd levels may be associated with advanced cancer phenotypes, but there was only a tenuous association between cadmium and prostate cancer. PMID- 20003242 TI - Molecular evolution of measles viruses circulated in Taiwan 1992-2008. AB - Genetic analyses of viral samples from 74 laboratory confirmed measles cases occurring in Taiwan during 1992-2008 identified six viral genotypes D3, D5, D9, G2, H1 and H2. The most frequently detected genotype, H1, was associated with outbreaks in 1994 and 2002, and was the likely indigenous genotype in 1992. In response to the outbreaks, two catch-up campaigns were launched and a routine second dose of measles, mumps, and rubella vaccine at entry to elementary school was introduced. The vaccination campaigns successfully reduced the number of measles cases in Taiwan, and many of the more recent cases can be traced to importations, primarily from other Asian countries. A number of measles genotypes which were associated with outbreaks in other Asian countries were detected among the more recent cases. The more recent genotype H1 viruses had sequences that were identical to those currently circulating in China or associated with international importation of virus. PMID- 20003243 TI - Gadolinium decreases inflammation related to myocardial ischemia and reperfusion injury. AB - BACKGROUND: The lanthanide cation, gadolinium (GdCl3) protects the myocardium against infarction following ischemia and reperfusion. Neutrophils and macrophages are the main leukocytes responsible for infarct expansion after reperfusion. GdCl3 interferes with macrophage and neutrophil function in the liver by decreasing macrophage secretion of inflammatory cytokines and neutrophil infiltration. We hypothesized that GdCl3 protects against ischemia and reperfusion injury by decreasing inflammation. We determined the impact of GdCl3 treatment for reperfusion injury on 1) circulating monoctye and neutrophil counts, 2) secretion of inflammatory cytokines, and 3) influx of monocytes and neutrophils into the myocardium. METHODS: Rats (n = 3-6/gp) were treated with saline or GdCl3 (20 mumol/kg) 15 min prior to a 30 min period of regional ischemia and 120 min reperfusion. Sham rats were not subject to ischemia. Blood was collected either after 30 min ischemia or 120 min reperfusion and hearts were harvested at 120 min reperfusion for tissue analysis. Blood was analyzed for leukocytes counts and cytokines. Tissue was analyzed for cytokines and markers of neutrophil and monocyte infiltration by measuring myeloperoxidase (MPO) and alpha naphthyl acetate esterase (ANAE). RESULTS: GdCl3 did not affect the number of circulating neutrophils prior to ischemia. Two hours reperfusion resulted in a 2- and 3- fold increase in circulating monocytes and neutrophils, respectively. GdCl3 decreased the number of circulating monocytes and neutrophils during reperfusion to levels below those present prior to ischemia. Furthermore, after 120 min of reperfusion, GdCl3 decreased ANAE and MPO activity in the myocardium by 1.9-fold and 6.5-fold respectively. GdCl3 decreased MPO activity to levels below those measured in the Sham group. Serum levels of the major neutrophil chemoattractant cytokine, IL-8 were increased from pre-ischemic levels during ischemia and reperfusion in both control and GdCl3 treated rats. Likewise, IL-8 levels increased throughout the 3 hour time period in the Sham group. There was no difference in IL-8 detected in the myocardium after 120 min reperfusion between groups. In contrast, after 120 min reperfusion GdCl3 decreased the myocardial tissue levels of macrophage secreted cytokines, GM-CSF and IL-1. CONCLUSION: GdCl3 treatment prior to ischemia and reperfusion injury decreased circulating monocytes and neutrophils, macrophage secreted cytokines, and leukocyte infiltration into injured myocardium. These results suggest GdCl3 decreased monoctye and neutrophil migration and activation and may be a novel treatment for inflammation during ischemia and reperfusion. PMID- 20003244 TI - Overexpression of inhibitor of DNA-binding (ID)-1 protein related to angiogenesis in tumor advancement of ovarian cancers. AB - BACKGROUND: The inhibitor of DNA-binding (ID) has been involved in cell cycle regulation, apoptosis and angiogenesis. This prompted us to study ID functions in tumor advancement of ovarian cancers. METHODS: Sixty patients underwent surgery for ovarian cancers. In ovarian cancers, the levels of ID-1, ID-2 and ID-3 mRNAs were determined by real-time reverse transcription-polymerase chain reaction. The histoscore with the localization of ID-1 was determined by immunohistochemistry. Patient prognosis was analyzed with a 36-month survival rate. Microvessel counts were determined by immunohistochemistry for CD34 and factor VIII-related antigen. RESULTS: ID-1 histoscores and mRNA levels both significantly (p < 0.001) increased in ovarian cancers according to clinical stage, regardless of histopathological type. Furthermore, 30 patients with high ID-1 expression had a lower survival rate (53%) compared to patients with low ID-1 expression (80%). ID 1 histoscores and mRNA levels significantly (p < 0.0001) correlated with microvessel counts in ovarian cancers. CONCLUSION: ID-1 increased in ovarian cancer cells during tumor progression. Moreover, ID-1 expression levels correlated with microvessel counts. Therefore, ID-1 might work on tumor advancement via angiogenesis and is considered to be a candidate for a prognostic indicator in ovarian cancers. PMID- 20003245 TI - Towards a comprehensive barcode library for arctic life - Ephemeroptera, Plecoptera, and Trichoptera of Churchill, Manitoba, Canada. AB - BACKGROUND: This study reports progress in assembling a DNA barcode reference library for Ephemeroptera, Plecoptera, and Trichoptera ("EPTs") from a Canadian subarctic site, which is the focus of a comprehensive biodiversity inventory using DNA barcoding. These three groups of aquatic insects exhibit a moderate level of species diversity, making them ideal for testing the feasibility of DNA barcoding for routine biotic surveys. We explore the correlation between the morphological species delineations, DNA barcode-based haplotype clusters delimited by a sequence threshold (2%), and a threshold-free approach to biodiversity quantification--phylogenetic diversity. RESULTS: A DNA barcode reference library is built for 112 EPT species for the focal region, consisting of 2277 COI sequences. Close correspondence was found between EPT morphospecies and haplotype clusters as designated using a standard threshold value. Similarly, the shapes of taxon accumulation curves based upon haplotype clusters were very similar to those generated using phylogenetic diversity accumulation curves, but were much more computationally efficient. CONCLUSION: The results of this study will facilitate other lines of research on northern EPTs and also bode well for rapidly conducting initial biodiversity assessments in unknown EPT faunas. PMID- 20003246 TI - IL4 gene polymorphism and previous malaria experiences manipulate anti-Plasmodium falciparum antibody isotype profiles in complicated and uncomplicated malaria. AB - BACKGROUND: The IL4-590 gene polymorphism has been shown to be associated with elevated levels of anti-Plasmodium falciparum IgG antibodies and parasite intensity in the malaria protected Fulani of West Africa. This study aimed to investigate the possible impact of IL4-590C/T polymorphism on anti-P. falciparum IgG subclasses and IgE antibodies levels and the alteration of malaria severity in complicated and uncomplicated malaria patients with or without previous malaria experiences. METHODS: Anti-P.falciparum IgG subclasses and IgE antibodies in plasma of complicated and uncomplicated malaria patients with or without previous malaria experiences were analysed using ELISA. IL4-590 polymorphisms were genotyped using RFLP-PCR. Statistical analyses of the IgG subclass levels were done by Oneway ANOVA. Genotype differences were tested by Chi-squared test. RESULTS: The IL4-590T allele was significantly associated with anti-P. falciparum IgG3 antibody levels in patients with complicated (P = 0.031), but not with uncomplicated malaria (P = 0.622). Complicated malaria patients with previous malaria experiences carrying IL4-590TT genotype had significantly lower levels of anti-P. falciparum IgG3 (P = 0.0156), while uncomplicated malaria patients with previous malaria experiences carrying the same genotype had significantly higher levels (P = 0.0206) compared to their IL4-590 counterparts. The different anti-P. falciparum IgG1 and IgG3 levels among IL4 genotypes were observed. Complicated malaria patients with previous malaria experiences tended to have lower IgG3 levels in individuals carrying TT when compared to CT genotypes (P = 0.075). In contrast, complicated malaria patients without previous malaria experiences carrying CC genotype had significantly higher anti-P. falciparum IgG1 than those carrying either CT or TT genotypes (P = 0.004, P = 0.002, respectively). CONCLUSION: The results suggest that IL4-590C or T alleles participated differently in the regulation of anti-malarial antibody isotype profiles in primary and secondary malaria infection and, therefore, could play an important role in alteration of malaria severity. PMID- 20003247 TI - Comparison of the eSwab collection and transportation system to an amies gel transystem for Gram stain of clinical specimens. AB - BACKGROUND: The first step in routine microbiology laboratory procedures is the collection and safe transportation of swab samples. This can be accomplished using ESwab Collection and Transport System (Copan Italia, Brescia -Italy). The aim of the present study was to compare the results of microscopic examination of gram stain smears prepared directly from clinical specimens, collected and transported in the ESwab, with those obtained using Amies Agar gel Transystem without charcoal (Copan). FINDINGS: Specimens were collected from 80 patients (32 vaginal swabs, 27 cervical swabs, 11 urethral swabs and 10 wound swabs). Two swabs were in random order collected from each patient, one using the conventional Amies gel Transystem, the other using ESwab. One slide was prepared for each specimen using the conventional swab and two sets of slides were prepared from the specimens collected with the ESwab: one using 100 mul and one using 50 mul of the Amies medium. All slides were gram stained using an automated Gram stainer. Microscopic examination of 240 slides (80 with conventional and 160 with ESwab) showed that the quality of smear preparation from the ESwab system, allowed for easier identification of human cells and identification of greater number of microorganisms. Microscopic examination of additional slides prepared from ESwab at 24 or 72 hours after initial collection were equivalent to those prepared when received in the laboratory within 2 hours of collection. CONCLUSION: Microscopic examination performed using ESwab, especially when preparing the slides with 100 mul, shows superior results to those obtained using the Amies gel Transystem. PMID- 20003248 TI - Outcomes following oesophagectomy in patients with oesophageal cancer: a secondary analysis of the ICNARC Case Mix Programme Database. AB - INTRODUCTION: This report describes the case mix and outcomes of patients with oesophageal cancer admitted to adult critical care units following elective oesophageal surgery in England, Wales and Northern Ireland. METHODS: Admissions to critical care following elective oesophageal surgery for malignancy were identified using data from the Intensive Care National Audit and Research Centre (ICNARC) Case Mix Programme Database. Information on admissions between December 1995 and September 2007 were extracted and the association between in-hospital mortality and patient characteristics on admission to critical care was assessed using multiple logistic regression analysis. The performance of three prognostic models (Simplified Acute Physiology Score (SAPS) II, Acute Physiology and Chronic Health Evaluation (APACHE) II and the ICNARC physiology score) was also evaluated. RESULTS: Between 1995 and 2007, there were 7227 admissions to 181 critical care units following oesophageal surgery for malignancy. Overall mortality in critical care was 4.4% and in-hospital mortality was 11%, although both declined steadily over time. Eight hundred and seventy-three (12.2%) patients were readmitted to critical care, most commonly for respiratory complications (49%) and surgical complications (25%). Readmitted patients had a critical care unit mortality of 24.7% and in-hospital mortality of 33.9%. Overall in-hospital mortality was associated with patient age, and various physiological measurements on admission to critical care (partial pressure of arterial oxygen (PaO2):fraction of inspired oxygen (FiO2) ratio, lowest arterial pH, mechanical ventilation, serum albumin, urea and creatinine). The three prognostic models evaluated performed poorly in measures of discrimination, calibration and goodness of fit. CONCLUSIONS: Surgery for oesophageal malignancy continues to be associated with significant morbidity and mortality. Age and organ dysfunction in the early postoperative period are associated with an increased risk of death. Postoperative serum albumin is confirmed as an additional prognostic factor. More work is required to determine how this knowledge may improve clinical management. PMID- 20003250 TI - Computer aided data acquisition tool for high-throughput phenotyping of plant populations. AB - BACKGROUND: The data generated during a course of a biological experiment/study can be sometimes be massive and its management becomes quite critical for the success of the investigation undertaken. The accumulation and analysis of such large datasets often becomes tedious for biologists and lab technicians. Most of the current phenotype data acquisition management systems do not cater to the specialized needs of large-scale data analysis. The successful application of genomic tools/strategies to introduce desired traits in plants requires extensive and precise phenotyping of plant populations or gene bank material, thus necessitating an efficient data acquisition system. RESULTS: Here we describe newly developed software "PHENOME" for high-throughput phenotyping, which allows researchers to accumulate, categorize, and manage large volume of phenotypic data. In this study, a large number of individual tomato plants were phenotyped with the "PHENOME" application using a Personal Digital Assistant (PDA) with built-in barcode scanner in concert with customized database specific for handling large populations. CONCLUSION: The phenotyping of large population of plants both in the laboratory and in the field is very efficiently managed using PDA. The data is transferred to a specialized database(s) where it can be further analyzed and catalogued. The "PHENOME" aids collection and analysis of data obtained in large-scale mutagenesis, assessing quantitative trait loci (QTLs), raising mapping population, sampling of several individuals in one or more ecological niches etc. PMID- 20003249 TI - Combination strategies for pandemic influenza response - a systematic review of mathematical modeling studies. AB - BACKGROUND: Individual strategies in pandemic preparedness plans may not reduce the impact of an influenza pandemic. METHODS: We searched modeling publications through PubMed and associated references from 1990 to 30 September 2009. Inclusion criteria were modeling papers quantifying the effectiveness of combination strategies, both pharmaceutical and non-pharmaceutical. RESULTS: Nineteen modeling papers on combination strategies were selected. Four studies examined combination strategies on a global scale, 14 on single countries, and one on a small community. Stochastic individual-based modeling was used in nine studies, stochastic meta-population modeling in five, and deterministic compartmental modeling in another five. As part of combination strategies, vaccination was explored in eight studies, antiviral prophylaxis and/or treatment in 16, area or household quarantine in eight, case isolation in six, social distancing measures in 10 and air travel restriction in six studies. Two studies suggested a high probability of successful influenza epicenter containment with combination strategies under favorable conditions. During a pandemic, combination strategies delayed spread, reduced overall number of cases, and delayed and reduced peak attack rate more than individual strategies. Combination strategies remained effective at high reproductive numbers compared with single strategy. Global cooperative strategies, including redistribution of antiviral drugs, were effective in reducing the global impact and attack rates of pandemic influenza. CONCLUSION: Combination strategies increase the effectiveness of individual strategies. They include pharmaceutical (antiviral agents, antibiotics and vaccines) and non-pharmaceutical interventions (case isolation, quarantine, personal hygiene measures, social distancing and travel restriction). Local epidemiological and modeling studies are needed to validate efficacy and feasibility. PMID- 20003251 TI - Knowledge translation research in population health: establishing a collaborative research agenda. AB - BACKGROUND: Despite the increasing mobilization of researchers and funding organizations around knowledge translation (KT) in Canada and elsewhere, many questions have been only partially answered, particularly in the field of population health. This article presents the results of a systematic process to draw out possible avenues of collaboration for researchers, practitioners and decision-makers who work in the area of KT. The main objective was to establish a research agenda on knowledge translation in population health. METHODS: Using the Concept Mapping approach, the research team wanted to identify priority themes for the development of research on KT in population health. Mapping is based on multivariate statistical analyses (multidimensional scaling and hierarchical cluster analysis) in which statements produced during a brainstorming session are grouped in weighted clusters. The final maps are a visual representation of the priority themes of research on KT. Especially designed for facilitating consensus in the understanding and organization of various concepts, the Concept Mapping method proved suitable for achieving this objective. RESULTS: The maps were produced by 19 participants from university settings, and from institutions within the health and social services network. Three main perspectives emerge from this operation: (1) The evaluation of the effectiveness of KT efforts is one of the main research priorities; (2) The importance of taking into consideration user contexts in any KT effort; (3) The challenges related to sharing power for decision-making and action-taking among various stakeholder groups. These perspectives open up avenues of collaboration for stakeholders who are involved in research on KT. Besides these three main perspectives, the concept maps reveal three other trends which should be emphasized. CONCLUSION: The Concept Mapping process reported in this article aimed to provoke collective reflection on the research questions that should be studied, in order to foster coherence in research activities in the field of population health. Based on this, it is appropriate to continue to support the development of research projects in KT and the formation of research teams in this field. Research on KT must lead to concrete outcomes within communities that are interested in the question. PMID- 20003252 TI - Carotid body tumors: radioguided surgical approach. AB - BACKGROUND: Carotid body tumours (CBTs) are very rare lesions which should be treated as soon as possible even when benign since small tumour size permits easier removal and lower incidence of perioperative complications and recurrence. Malignant forms are rare and they can be identified by lymph node invasion and metastases in distant locations. The need of reliable and effective diagnostic modalities for both primary CBTs and its metastases or recurrence is evident.The present study reviews our experience and attempt to define the role of colour coded ultrasound (CCU) and Somatostatin receptor scintigraphy (SRS) with Indium 111-DTPA-pentetretide (Octreoscan(R)) using both planar and single photon emission tomography (SPECT) technique in the diagnosis and follow-up of these uncommon lesions within a multidisciplinary approach. METHODS: From 1997 to 2008, 12 patients suffering from 16 CBTs (4 bilateral) were investigated by CCU and SRS SPECT before and after surgery. All tumours were grouped according to Shamblin's classification in order to assess the technical difficulties and morbidity of surgical resection on the ground of their size and relationship with the carotid arteries. Intraoperative radiocaptation by Octreoscan(R)) was also carried out in all cases to evaluate the radicality of surgery. All perioperative scans were evaluated by the same nuclear medicine physician. RESULTS: Preoperatively CCU showed CBTs (four were not palpable) with a sensitivity of 100%. Radioisotope imaging identified the CBTs as chemodectomas in 15 cases while no radioisotopic uptake was detected in 1 vagus nerve neurinoma. No evidence of metastasis or multicentricity were seen by total body radioisotopic scans. Combined data from CCU and SRS-SPECT allowed to determine tumour size in order to select 7 larger tumours which were submitted to selective preoperative embolization.Intraoperatively Octreoscan demonstrated microscopic tumour leftovers promptly removed in 1 case and an unresectable remnant at the base of the skull in another case.During follow-up CCI and radioisotope scans showed no recurrence in 14 cases and a slightly enlargement of the intracranial residual as detected during surgery in 1 patient. CONCLUSION: CCU may allow an early and noninvasive detection of CBTs and hence safer operations. The combined use of CCU and SRS-SPECT provide useful data to identify those tumours and to evaluate their extent and carotid arteries infiltration. Radioisotope imaging is a sensitive modality to detect metastases and lymph node involvement that are markers of CBT malignancy. After surgery CCU and SRS-SPECT can be accurate modalities for surveillance for an early detection of CBTs recurrence. PMID- 20003253 TI - Effects of chromium picolinate on glycemic control and kidney of the obese Zucker rat. AB - BACKGROUND: Chromium picolinate (Cr(pic)3) is advocated as adjuvant therapy for impaired glycemic control, despite concerns for DNA damage. Potential toxicity of Cr(pic)3 should be greater for the kidney that accumulates chromium. Therefore, we tested the hypothesis that Cr(pic)3 treatment of obese Zucker rats (OZR) exacerbates renal abnormalities associated with dysglycemia. METHODS: Male OZR were treated with diets lacking or containing 5 and 10 mg/kg of chromium, as Cr(pic)3, for 20 weeks; lean Zucker rats (LZR) served as controls. Glycemic and renal effects of Cr(pic)3 were determined in the context of indices of oxidative stress and inflammation. RESULTS: The OZR displayed increased fasting plasma glucose and insulin in association with enlarged pancreatic islets exhibiting collagen and periodic acid Schiff-positive deposits compared to LZR; Cr(pic)3 treatment did not affect these parameters. The OZR, irrespective of Cr(pic)3, excreted more albumin than LZR. Also, other indices of renal function or histopathology were not affected by Cr(pic)3 treatment. Urinary excretion of 8 hydroxydeoxyguanosine (8-OHdG), an index of oxidative DNA damage, was greater in the OZR than LZR; dietary Cr(pic)3 treatment attenuated 8-OHdG excretion. However, immunostaining of kidney for 8-OHdG revealed similar staining pattern and intensity, despite significant renal accumulation of chromium in Cr(pic)3 treated groups. Finally, increased renal nitrotyrosine and cyclooxygenase-2 levels and urinary excretion of monocyte chemoattractant protein-1 of OZR were partially reversed by Cr(pic)3 treatment. CONCLUSION: Dietary Cr(pic)3 treatment of OZR does not beneficially influence glycemic status or increase the risk for oxidative DNA damage; rather, the treatment attenuates indices of oxidative stress and inflammation. PMID- 20003254 TI - Motor function may differentiate attention deficit hyperactivity disorder from early onset bipolar disorder. AB - BACKGROUND: Differentiating between bipolar spectrum disorder (BD) and attention deficit hyperactivity disorder (ADHD) in childhood and adolescence is difficult because the clinical presentation is influenced by ongoing neural development, causing considerable symptom overlap. Motor problems and neurological soft signs have been associated with ADHD for decades. Little is known about motor skills in BD. Here we assess the diagnostic accuracy of neuromotor deviations in differentiating ADHD from BD in clinical practice. We also investigate if these deviations exist in concurrent ADHD and BD, thus indicating true comorbidity METHODS: 64 patients 6-18 years (31 girls, 33 boys) fulfilling the diagnostic criteria of BD, ADHD combined subtype (ADHD-C) or comorbid BD and ADHD-C, were compared using an age-standardized neuromotor test; NUBU. Categorical variables were analyzed using cross table with two-tailed chi square test or Fisher's exact test when appropriate. Continuous variables were analyzed by Kruskal-Wallis test and, if significant, Mann-Whitney U test and ROC plots. RESULTS: The ADHD-C group and the comorbid ADHD-C and BD group both showed significantly more neurological soft signs (p less than 0.01) and lower mean static coordination percentile (p less than 0.01) than the BD group. The positive predictive value of NUBU in the diagnosis of ADHD-C with or without concurrent BD was 89% (80-95) for total soft signs and 87% (79-95) for static coordination below the 7.5 percentile. CONCLUSION: An age-standardized neuromotor test battery may promote diagnostic accuracy in differentiating ADHD from BD in clinical practice, and help evaluating whether symptoms of ADHD in children who have BD reflect symptom overlap or real comorbidity. This may have important implications for everyday diagnostic work. PMID- 20003255 TI - Synthesis and expression of CDw75 antigen in human colorectal cancer. AB - BACKGROUND: Increased ST6Gal I activity has been associated with the alpha(2,6)sialylation enhancement of membrane glycoconjugates observed in metastatic colorectal carcinomas (CRC). Siaalpha(2,6)Galbeta(1,4)GlcNAc sequence, known as CDw75, is a sialylated carbohydrate determinant generated by the ST6Gal I. This epitope has been reported to be associated with the progression of gastric and colorectal tumours, hence there are only a few conclusive studies to date. METHODS: By radioisotopic techniques we evaluated the ST6Gal I activity in healthy, transitional and tumour tissues from 43 patients with CRC. By immunohistochemistry we assessed the CDw75 expression in 25 colorectal adenomas, 43 tumours, 13 transitional and 28 healthy tissues of CRC patients. RESULTS: ST6Gal I activity was likewise found to be statistically higher in tumour tissue respect to healthy tissue from CRC patients. CDw75 expression was positive in 20% of colorectal adenomas. Furthermore, 70% of tumour specimens and 8.3% of transitional specimens were positive for CDw75 expression, whereas none of the healthy ones showed the presence of the epitope. CONCLUSION: The major contribution of this study is the inclusion of data from transitional tissue and the analysis of CDw75 antigen expression in CRC and in colorectal adenomas, little known so far. ST6Gal I activity and CDw75 antigen expression were increased in CRC. Although their comparison did not reach the statistical significance, a great extent of patients showed both, an enhanced tumour ST6Gal I activity and an increased CDw75 expression in the tumour tissue. So, these two variables may play a role in malignant transformation. The expression of CDw75 in colorectal adenomas suggests that this antigen may be a tumour marker in CRC. PMID- 20003256 TI - Validation of computerized diagnostic information in a clinical database from a national equine clinic network. AB - BACKGROUND: Computerized diagnostic information offers potential for epidemiological research; however data accuracy must be addressed. The principal aim of this study was to evaluate the completeness and correctness of diagnostic information in a computerized equine clinical database compared to corresponding hand written veterinary clinical records, used as gold standard, and to assess factors related to correctness. Further, the aim was to investigate completeness (epidemiologic sensitivity), correctness (positive predictive value), specificity and prevalence for diagnoses for four body systems and correctness for affected limb information for four joint diseases. METHODS: A random sample of 450 visits over the year 2002 (nvisits=49,591) was taken from 18 nation wide clinics headed under one company. Computerized information for the visits selected and copies of the corresponding veterinary clinical records were retrieved. Completeness and correctness were determined using semi-subjective criteria. Logistic regression was used to examine factors associated with correctness for diagnosis. RESULTS: Three hundred and ninety six visits had veterinary clinical notes that were retrievable. The overall completeness and correctness were 91% and 92%, respectively; both values considered high. Descriptive analyses showed significantly higher degree of correctness for first visits compared to follow up visits and for cases with a diagnostic code recorded in the veterinary records compared to those with no code noted. The correctness was similar regardless of usage category (leisure/sport horse, racing trotter and racing thoroughbred) or gender.For the four body systems selected (joints, skin and hooves, respiratory, skeletal) the completeness varied between 71% (respiration) and 91% (joints) and the correctness ranged from 87% (skin and hooves) to 96% (respiration), whereas the specificity was >95% for all systems. Logistic regression showed that correctness was associated with type of visit, whether an explicit diagnostic code was present in the veterinary clinical record, and body system. Correctness for information on affected limb was 95% and varied with joint. CONCLUSION: Based on the overall high level of correctness and completeness the database was considered useful for research purposes. For the body systems investigated the highest level of completeness and correctness was seen for joints and respiration, respectively. PMID- 20003257 TI - Cryptic Leishmania infantum infection in Italian HIV infected patients. AB - BACKGROUND: Visceral leishmaniasis (VL) is a protozoan diseases caused in Europe by Leishmania (L.) infantum. Asymptomatic Leishmania infection is more frequent than clinically apparent disease. Among HIV infected patients the risk of clinical VL is increased due to immunosuppression, which can reactivate a latent infection. The aims of our study were to assess the prevalence of asymptomatic L. infantum infection in HIV infected patients and to study a possible correlation between Leishmania parasitemia and HIV infection markers. METHODS: One hundred and forty-five HIV infected patients were screened for the presence of anti Leishmania antibodies and L. infantum DNA in peripheral blood. Statistical analysis was carried out by using a univariate regression analysis. RESULTS: Antibodies to L. infantum were detected in 1.4% of patients. L. infantum DNA was detected in 16.5% of patients. Significant association for PCR-Leishmania levels with plasma viral load was documented (p = 0.0001). CONCLUSION: In our area a considerable proportion of HIV infected patients are asymptomatic carriers of L. infantum infection. A relationship between high HIV viral load and high parasitemic burden, possibly related to a higher risk of developing symptomatic disease, is suggested. PCR could be used for periodic screening of HIV patients to individuate those with higher risk of reactivation of L. infantum infection. PMID- 20003258 TI - Comparative analysis of catfish BAC end sequences with the zebrafish genome. AB - BACKGROUND: Comparative mapping is a powerful tool to transfer genomic information from sequenced genomes to closely related species for which whole genome sequence data are not yet available. However, such an approach is still very limited in catfish, the most important aquaculture species in the United States. This project was initiated to generate additional BAC end sequences and demonstrate their applications in comparative mapping in catfish. RESULTS: We reported the generation of 43,000 BAC end sequences and their applications for comparative genome analysis in catfish. Using these and the additional 20,000 existing BAC end sequences as a resource along with linkage mapping and existing physical map, conserved syntenic regions were identified between the catfish and zebrafish genomes. A total of 10,943 catfish BAC end sequences (17.3%) had significant BLAST hits to the zebrafish genome (cutoff value 98%) were mild to moderate in intensity, and the most commonly reported AEs included headache, decreased appetite, insomnia, and abdominal pain. Seven subjects discontinued the study due to intolerable AEs (n = 3) and application site reactions (n = 4). CONCLUSION: This study demonstrates that MTS, when carefully titrated to optimal dose, may further improve child and family HRQL, as well as behavioral, medication worry, and economic impact item scores, as measured by the AIM-C in subjects switching to MTS from a stable dose of routinely prescribed oral ER-MPH after a short treatment period. Furthermore, following the abrupt conversion from oral ER-MPH to MTS, the majority of caregivers reported being highly satisfied with MTS as a treatment option for their children with ADHD. TRIAL REGISTRATION: NCT00151983. PMID- 20003261 TI - Optimized PCR with sequence specific primers (PCR-SSP) for fast and efficient determination of Interleukin-6 Promoter -597/-572/-174Haplotypes. AB - BACKGROUND: Interleukin-6 (IL-6) promoter polymorphisms at positions -597(G-->A), -572(G-->C) and -174(G-->C) were shown to have a clinical impact on different major diseases. At present PCR-SSP protocols for IL-6 -597/-572/-174haplotyping are elaborate and require large amounts of genomic DNA. FINDINGS: We describe an improved typing technique requiring a decreased number of PCR-reactions and a reduced PCR-runtime due to optimized PCR-conditions. CONCLUSION: This enables a fast and efficient determination of IL-6 -597/-572/-174haplotypes in clinical diagnosis and further evaluation of IL-6 promoter polymorphisms in larger patient cohorts. PMID- 20003262 TI - Effect of quinolinic acid on human astrocytes morphology and functions: implications in Alzheimer's disease. AB - The excitotoxin quinolinic acid (QUIN) is synthesized through the kynurenine pathway (KP) by activated monocyte lineage cells. QUIN is likely to play a role in the pathogenesis of several major neuroinflammatory diseases including Alzheimer's disease (AD). The presence of reactive astrocytes, astrogliosis, increased oxidative stress and inflammatory cytokines are important pathological hallmarks of AD. We assessed the stimulatory effects of QUIN at low physiological to high excitotoxic concentrations in comparison with the cytokines commonly associated with AD including IFN-gamma and TNF-alpha on primary human astrocytes. We found that QUIN induces IL-1beta expression, a key mediator in AD pathogenesis, in human astrocytes. We also explored the effect of QUIN on astrocyte morphology and functions. At low concentrations, QUIN treatment induced concomitantly a marked increase in glial fibrillary acid protein levels and reduction in vimentin levels compared to controls; features consistent with astrogliosis. At pathophysiological concentrations QUIN induced a switch between structural protein expressions in a dose dependent manner, increasing VIM and concomitantly decreasing GFAP expression. Glutamine synthetase (GS) activity was used as a functional metabolic test for astrocytes. We found a significant dose dependent reduction in GS activity following QUIN treatment. All together, this study showed that QUIN is an important factor for astroglial activation, dysregulation and cell death with potential relevance to AD and other neuroinflammatory diseases. PMID- 20003263 TI - Invasions, DNA barcodes, and rapid biodiversity assessment using ants of Mauritius. AB - BACKGROUND: Using an understudied taxon (Hymenoptera, Formicidae) found on a tropical island (Mauritius) where native flora and fauna have been threatened by 400 years of habitat modification and introduced species, we tested whether estimated incidences of diversity and complementarity were similar when measured by standard morphological alpha-taxonomy or phylogenetic diversity (PD) based on a standardized mitochondrial barcode and corroborating nuclear marker. RESULTS: We found that costs related to site loss (considered loss of evolutionary history measured as loss of barcode PD) were not significantly different from predictions made either a) using standard morphology-based taxonomy, or b) measured using a nuclear marker. Integrating morphology and barcode results permitted us to identify a case of initially morphologically-cryptic variation as a new and endemic candidate species. However, barcode estimates of the relative importance of each site or network of sites were dramatically affected when the species in question was known to be indigenous or introduced. CONCLUSION: This study goes beyond a mere demonstration of the rapid gains possible for diversity assessment using a standardized DNA barcode. Contextualization of these gains with ecological and natural history information is necessary to calibrate this wealth of standardized information. Without such an integrative approach, critical opportunities to advance knowledge will be missed. PMID- 20003264 TI - Intein-mediated site-specific conjugation of Quantum Dots to proteins in vivo. AB - We describe an intein based method to site-specifically conjugate Quantum Dots (QDs) to target proteins in vivo. This approach allows the covalent conjugation of any nanostructure and/or nanodevice to any protein and thus the targeting of such material to any intracellular compartment or signalling complex within the cells of the developing embryo. We genetically fused a pleckstrin-homology (PH) domain with the N-terminus half of a split intein (IN). The C-terminus half (IC) of the intein was conjugated to QDs in vitro. IC-QD's and RNA encoding PH-IN were microinjected into Xenopus embryos. In vivo intein-splicing resulted in fully functional QD-PH conjugates that could be monitored in real time within live embryos. Use of Near Infra Red (NIR)-emitting QDs allowed monitoring of QD conjugates within the embryo at depths where EGFP is undetectable demonstrating the advantages of QD's for this type of experiment. In conclusion, we have developed a novel in vivo methodology for the site-specific conjugation of QD's and other artificial structures to target proteins in different intracellular compartments and signaling complexes. PMID- 20003265 TI - Extent of differential allelic expression of candidate breast cancer genes is similar in blood and breast. AB - INTRODUCTION: Normal gene expression variation is thought to play a central role in inter-individual variation and susceptibility to disease. Regulatory polymorphisms in cis-acting elements result in the unequal expression of alleles. Differential allelic expression (DAE) in heterozygote individuals could be used to develop a new approach to discover regulatory breast cancer susceptibility loci. As access to large numbers of fresh breast tissue to perform such studies is difficult, a suitable surrogate test tissue must be identified for future studies. METHODS: We measured differential allelic expression of 12 candidate genes possibly related to breast cancer susceptibility (BRCA1, BRCA2, C1qA, CCND3, EMSY, GPX1, GPX4, MLH3, MTHFR, NBS1, TP53 and TRXR2) in breast tissue (n = 40) and fresh blood (n = 170) of healthy individuals and EBV-transformed lymphoblastoid cells (n = 19). Differential allelic expression ratios were determined by Taqman assay. Ratio distributions were compared using t-test and Wilcoxon rank sum test, for mean ratios and variances respectively. RESULTS: We show that differential allelic expression is common among these 12 candidate genes and is comparable between breast and blood (fresh and transformed lymphoblasts) in a significant proportion of them. We found that eight out of nine genes with DAE in breast and fresh blood were comparable, as were 10 out of 11 genes between breast and transformed lymphoblasts. CONCLUSIONS: Our findings support the use of differential allelic expression in blood as a surrogate for breast tissue in future studies on predisposition to breast cancer. PMID- 20003266 TI - The use of mobile phone data for the estimation of the travel patterns and imported Plasmodium falciparum rates among Zanzibar residents. AB - BACKGROUND: Malaria endemicity in Zanzibar has reached historically low levels, and the epidemiology of malaria transmission is in transition. To capitalize on these gains, Zanzibar has commissioned a feasibility assessment to help inform on whether to move to an elimination campaign. Declining local transmission has refocused attention on imported malaria. Recent studies have shown that anonimized mobile phone records provide a valuable data source for characterizing human movements without compromising the privacy of phone users. Such movement data in combination with spatial data on P. falciparum endemicity provide a way of characterizing the patterns of parasite carrier movements and the rates of malaria importation, which have been used as part of the malaria elimination feasibility assessment for the islands of Zanzibar. DATA AND METHODS: Records encompassing three months of complete mobile phone usage for the period October December 2008 were obtained from the Zanzibar Telecom (Zantel) mobile phone network company, the principal provider on the islands of Zanzibar. The data included the dates of all phone usage by 770,369 individual anonymous users. Each individual call and message was spatially referenced to one of six areas: Zanzibar and five mainland Tanzania regions. Information on the numbers of Zanzibar residents travelling to the mainland, locations visited and lengths of stay were extracted. Spatial and temporal data on P. falciparum transmission intensity and seasonality enabled linkage of this information to endemicity exposure and, motivated by malaria transmission models, estimates of the expected patterns of parasite importation to be made. RESULTS: Over the three month period studied, 88% of users made calls that were routed only through masts on Zanzibar, suggesting that no long distance travel was undertaken by this group. Of those who made calls routed through mainland masts the vast majority of trips were estimated to be of less than five days in length, and to the Dar Es Salaam Zantel defined region. Though this region covered a wide range of transmission intensities, data on total infection numbers in Zanzibar combined with mathematical models enabled informed estimation of transmission exposure and imported infection numbers. These showed that the majority of trips made posed a relatively low risk for parasite importation, but risk groups visiting higher transmission regions for extended periods of time could be identified. CONCLUSION: Anonymous mobile phone records provide valuable information on human movement patterns in areas that are typically data-sparse. Estimates of human movement patterns from Zanzibar to mainland Tanzania suggest that imported malaria risk from this group is heterogeneously distributed; a few people account for most of the risk for imported malaria. In combination with spatial data on malaria endemicity and transmission models, movement patterns derived from phone records can inform on the likely sources and rates of malaria importation. Such information is important for assessing the feasibility of malaria elimination and planning an elimination campaign. PMID- 20003267 TI - Exploring complex miRNA-mRNA interactions with Bayesian networks by splitting averaging strategy. AB - BACKGROUND: microRNAs (miRNAs) regulate target gene expression by controlling their mRNAs post-transcriptionally. Increasing evidence demonstrates that miRNAs play important roles in various biological processes. However, the functions and precise regulatory mechanisms of most miRNAs remain elusive. Current research suggests that miRNA regulatory modules are complicated, including up-, down-, and mix-regulation for different physiological conditions. Previous computational approaches for discovering miRNA-mRNA interactions focus only on down-regulatory modules. In this work, we present a method to capture complex miRNA-mRNA interactions including all regulatory types between miRNAs and mRNAs. RESULTS: We present a method to capture complex miRNA-mRNA interactions using Bayesian network structure learning with splitting-averaging strategy. It is designed to explore all possible miRNA-mRNA interactions by integrating miRNA-targeting information, expression profiles of miRNAs and mRNAs, and sample categories. We also present an analysis of data sets for epithelial and mesenchymal transition (EMT). Our results show that the proposed method identified all possible types of miRNA-mRNA interactions from the data. Many interactions are of tremendous biological significance. Some discoveries have been validated by previous research, for example, the miR-200 family negatively regulates ZEB1 and ZEB2 for EMT. Some are consistent with the literature, such as LOX has wide interactions with the miR-200 family members for EMT. Furthermore, many novel interactions are statistically significant and worthy of validation in the near future. CONCLUSIONS: This paper presents a new method to explore the complex miRNA-mRNA interactions for different physiological conditions using Bayesian network structure learning with splitting-averaging strategy. The method makes use of heterogeneous data including miRNA-targeting information, expression profiles of miRNAs and mRNAs, and sample categories. Results on EMT data sets show that the proposed method uncovers many known miRNA targets as well as new potentially promising miRNA-mRNA interactions. These interactions could not be achieved by the normal Bayesian network structure learning. PMID- 20003268 TI - The effect of enteral and parenteral feeding on secretion of orexigenic peptides in infants. AB - BACKGROUND: The feeding in the first months of the life seems to influence the risks of obesity and affinity to some diseases including atherosclerosis. The mechanisms of these relations are unknown, however, the modification of hormonal action can likely be taken into account. Therefore, in this study the levels of ghrelin and orexin A - peripheral and central peptide from the orexigenic gut brain axis were determined. METHODS: Fasting and one hour after the meal plasma concentrations of ghrelin and orexin were measured in breast-fed (group I; n = 17), milk formula-fed (group II; n = 16) and highly hydrolyzed, hypoallergic formula-fed (group III; n = 14) groups, age matched infants (mean 4 months) as well as in children with iv provision of nutrients (glucose - group IV; n = 15; total parenteral nutrition - group V; n = 14). Peptides were determined using EIA commercial kits. RESULTS: Despite the similar caloric intake in orally fed children the fasting ghrelin and orexin levels were significantly lower in the breast-fed children (0.37 +/- 0.17 and 1.24 +/- 0.29 ng/ml, respectively) than in the remaining groups (0.5 +/- 0.27 and 1.64 +/- 0.52 ng/ml, respectively in group II and 0.77 +/- 0.27 and 2.04 +/- 1.1 ng/ml, respectively, in group III). The postprandial concentrations of ghrelin increased to 0.87 +/- 0.29 ng/ml, p < 0.002 and 0.76 +/- 0.26 ng/ml, p < 0.01 in groups I and II, respectively as compared to fasting values. The decrease in concentration of ghrelin after the meal was observed only in group III (0.47 +/- 0.24 ng/ml). The feeding did not influence the orexin concentration. In groups IV and V the ghrelin and orexin levels resembled those in milk formula-fed children. CONCLUSION: The highly hydrolyzed diet strongly affects fasting and postprandial ghrelin and orexin plasma concentrations with possible negative effect on short- and long-time effects on development. Also total parenteral nutrition with the continuous stimulation and lack of fasting/postprandial modulation might be responsible for disturbed development in children fed this way. PMID- 20003269 TI - Pharmacokinetics and lung delivery of PDDS-aerosolized amikacin (NKTR-061) in intubated and mechanically ventilated patients with nosocomial pneumonia. AB - INTRODUCTION: Aminoglycosides aerosolization might achieve better diffusion into the alveolar compartment than intravenous use. The objective of this multicenter study was to evaluate aerosol-delivered amikacin penetration into the alveolar epithelial lining fluid (ELF) using a new vibrating mesh nebulizer (Pulmonary Drug Delivery System (PDDS), Nektar Therapeutics), which delivers high doses to the lungs. METHODS: Nebulized amikacin (400 mg bid) was delivered to the lungs of 28 mechanically ventilated patients with Gram-negative VAP for 7-14 days, adjunctive to intravenous therapy. On treatment day 3, 30 minutes after completing aerosol delivery, all the patients underwent bronchoalveolar lavage in the infection-involved area and the ELF amikacin concentration was determined. The same day, urine and serum amikacin concentrations were determined at different time points. RESULTS: Median (range) ELF amikacin and maximum serum amikacin concentrations were 976.1 (135.7-16127.6) and 0.9 (0.62-1.73) microg/mL, respectively. The median total amount of amikacin excreted in urine during the first and second 12-hour collection on day 3 were 19 (12.21-28) and 21.2 (14.1 29.98) microg, respectively. During the study period, daily through amikacin measurements were below the level of nephrotoxicity. Sixty-four unexpected adverse events were reported, among which 2 were deemed possibly due to nebulized amikacin: one episode of worsening renal failure, and one episode of bronchospasm. CONCLUSIONS: PDDS delivery of aerosolized amikacin achieved very high aminoglycoside concentrations in ELF from radiography-controlled infection involved zones, while maintaining safe serum amikacin concentrations. The ELF concentrations always exceeded the amikacin minimum inhibitory concentrations for Gram-negative microorganisms usually responsible for these pneumonias. The clinical impact of amikacin delivery with this system remains to be determined. TRIAL REGISTRATION: ClinicalTrials.gov Identifier: NCT01021436. PMID- 20003271 TI - Expression of HIF-1alpha and VEGF in colorectal cancer: association with clinical outcomes and prognostic implications. AB - BACKGROUND: Hypoxia-inducible factor 1 alpha (HIF-1alpha) and vascular endothelial growth factor (VEGF) are frequently overexpressed in numerous types of cancers and are known to be important regulators of angiogenesis. Until now, few studies have been carried out to investigate the prognostic role of these factors in solid tumors, especially in colorectal cancer (CRC). The purpose of this study was to evaluate the expression of HIF-1alpha and VEGF in CRC tissues, and to analyze the association of these two factors with several clinical and pathological characteristics, and patients' survival. METHODS: Paraffin-embedded tissue samples were retrospectively collected from 71 CRC patients, who received surgical resection between 2001 and 2002, with a median follow-up of 5 years. We examined the patterns of expression of HIF-1alpha and VEGF by immunohistochemistry method. Statistical analysis was performed with univariate tests and multivariate Cox proportional hazards model to evaluate the differences. RESULTS: Expression of HIF-1alpha and VEGF was positively observed in 54.93% and 56.34% among the patients, respectively. HIF-1alpha and VEGF status were significantly associated with tumor stage, lymph nodes and liver metastases (P < 0.05). Expression of both HIF-1alpha and VEGF remained significantly associated with overall survival (OS) (P < 0.01), and HIF-1alpha was positively correlative to VEGF in CRC (r = 0.72, P < 0.001). CONCLUSIONS: HIF-1alpha and VEGF could be used as biomarkers indicating tumors in advanced stage and independently implied poor prognosis in patients with CRC. Treatment that inhibits HIF-1alpha might be a promising targeted approach in CRC to exhibit its potential to improve outcomes in future perspective, just as VEGF targeting has proved to be. PMID- 20003270 TI - Adhesive threads of extraintestinal pathogenic Escherichia coli. AB - The ability to adhere to host surfaces is by far the most vital step in the successful colonization by microbial pathogens. Colonization begins with the attachment of the bacterium to receptors expressed by cells forming the lining of the mucosa. Long hair like extracellular appendages called fimbriae, produced by most Gram-negative pathogens, mediate specific attachment to the epithelial cell surface. Associated with the fimbriae is a protein called an adhesin, which directs high-affinity binding to specific cell surface components. In the last couple of years, an enormous amount of research has been undertaken that deals with understanding how bacterial pathogens adhere to host cells. E. coli in all probability is one of the best studied free-living organisms. A group of E. coli called Extraintestinal pathogenic E. coli (ExPEC) including both human and animal pathogens like Uropathogenic E. coli (UPEC), Newborn meningitic E. coli (NMEC) and Avian pathogenic E. coli (APEC), have been found to harbour many fimbriae including Type 1 fimbriae, P fimbriae, curli fibres, S fimbriae, F1C fimbriae, Dr fimbriae, afimbrial adhesins, temperature-sensitive haemagglutinin and many novel adhesin gene clusters that have not yet been characterized. Each of these adhesins is unique due to the recognition of an adhesin-specific receptor, though as a group these adhesins share common genomic organization. A newly identified putative adhesin temporarily termed ExPEC Adhesin I, encoded by gene yqi, has been recently found to play a significant role in the pathogenesis of APEC infection, thus making it an interesting candidate for future research. The aim of this review is to describe the role of ExPEC adhesins during extraintestinal infections known till date, and to suggest the idea of investigating their potential role in the colonization of the host gut which is said to be a reservoir for ExPEC. PMID- 20003272 TI - CENPA overexpression promotes genome instability in pRb-depleted human cells. AB - BACKGROUND: Aneuploidy is a hallmark of most human cancers that arises as a consequence of chromosomal instability and it is frequently associated with centrosome amplification. Functional inactivation of the Retinoblastoma protein (pRb) has been indicated as a cause promoting chromosomal instability as well centrosome amplification. However, the underlying molecular mechanism still remains to be clarified. RESULTS: Here we show that pRb depletion both in wild type and p53 knockout HCT116 cells was associated with the presence of multipolar spindles, anaphase bridges, lagging chromosomes and micronuclei harbouring whole chromosomes. In addition aneuploidy caused by pRb acute loss was not affected by p53 loss.Quantitative real-time RT-PCR showed that pRB depletion altered expression of genes involved in centrosome duplication, kinetochore assembly and in the Spindle Assembly Checkpoint (SAC). However, despite MAD2 up-regulation pRb depleted cells seemed to have a functional SAC since they arrested in mitosis after treatments with mitotic poisons. Moreover pRb-depleted HCT116 cells showed BRCA1 overexpression that seemed responsible for MAD2 up-regulation.Post transcriptional silencing of CENPA by RNA interference, resulting in CENP-A protein levels similar to those present in control cells greatly reduced aneuploid cell numbers in pRb-depleted cells. CONCLUSION: Altogether our findings indicate a novel aspect of pRb acute loss that promotes aneuploidy mainly by inducing CENPA overexpression that in turn might induce micronuclei by affecting the correct attachment of spindle microtubules to kinetochores. PMID- 20003273 TI - The low-affinity phosphate transporter PitA is dispensable for in vitro growth of Mycobacterium smegmatis. AB - BACKGROUND: Mycobacteria have been shown to contain an apparent redundancy of high-affinity phosphate uptake systems, with two to four copies of such systems encoded in all mycobacterial genomes sequenced to date. In addition, all mycobacteria also contain at least one gene encoding the low-affinity phosphate transporter, Pit. No information is available on a Pit system from a Gram positive microorganism, and the importance of this system in a background of multiple other phosphate transporters is unclear. RESULTS: The aim of this study was to determine the physiological role of the PitA phosphate transporter in Mycobacterium smegmatis. Expression of pitA was found to be constitutive under a variety of growth conditions. An unmarked deletion mutant in pitA of M. smegmatis was created. The deletion did not affect in vitro growth or phosphate uptake of M. smegmatis. Expression of the high-affinity transporters, PstSCAB and PhnDCE, was increased in the pitA deletion strain. CONCLUSION: PitA is the only low affinity phosphate transport system annotated in the genome of M. smegmatis. The lack of phenotype of the pitA deletion strain shows that this system is dispensable for in vitro growth of this organism. However, increased expression of the remaining phosphate transporters in the mutant indicates a compensatory mechanism and implies that PitA is indeed used for the uptake of phosphate in M. smegmatis. PMID- 20003274 TI - Comparison of linear discriminant analysis methods for the classification of cancer based on gene expression data. AB - BACKGROUND: More studies based on gene expression data have been reported in great detail, however, one major challenge for the methodologists is the choice of classification methods. The main purpose of this research was to compare the performance of linear discriminant analysis (LDA) and its modification methods for the classification of cancer based on gene expression data. METHODS: The classification performance of linear discriminant analysis (LDA) and its modification methods was evaluated by applying these methods to six public cancer gene expression datasets. These methods included linear discriminant analysis (LDA), prediction analysis for microarrays (PAM), shrinkage centroid regularized discriminant analysis (SCRDA), shrinkage linear discriminant analysis (SLDA) and shrinkage diagonal discriminant analysis (SDDA). The procedures were performed by software R 2.80. RESULTS: PAM picked out fewer feature genes than other methods from most datasets except from Brain dataset. For the two methods of shrinkage discriminant analysis, SLDA selected more genes than SDDA from most datasets except from 2-class lung cancer dataset. When comparing SLDA with SCRDA, SLDA selected more genes than SCRDA from 2-class lung cancer, SRBCT and Brain dataset, the result was opposite for the rest datasets. The average test error of LDA modification methods was lower than LDA method. CONCLUSIONS: The classification performance of LDA modification methods was superior to that of traditional LDA with respect to the average error and there was no significant difference between theses modification methods. PMID- 20003275 TI - Researching the mental health needs of hard-to-reach groups: managing multiple sources of evidence. AB - BACKGROUND: Common mental health problems impose substantial challenges to patients, carers, and health care systems. A range of interventions have demonstrable efficacy in improving the lives of people experiencing such problems. However many people are disadvantaged, either because they are unable to access primary care, or because access does not lead to adequate help. New methods are needed to understand the problems of access and generate solutions. In this paper we describe our methodological approach to managing multiple and diverse sources of evidence, within a research programme to increase equity of access to high quality mental health services in primary care. METHODS: We began with a scoping review to identify the range and extent of relevant published material, and establish key concepts related to access. We then devised a strategy to collect - in parallel - evidence from six separate sources: a systematic review of published quantitative data on access-related studies; a meta-synthesis of published qualitative data on patient perspectives; dialogues with local stakeholders; a review of grey literature from statutory and voluntary service providers; secondary analysis of patient transcripts from previous qualitative studies; and primary data from interviews with service users and carers.We synthesised the findings from these diverse sources, made judgements on key emerging issues in relation to needs and services, and proposed a range of potential interventions. These proposals were debated and refined using iterative electronic and focus group consultation procedures involving international experts, local stakeholders and service users. CONCLUSIONS: Our methods break new ground by generating and synthesising multiple sources of evidence, connecting scientific understanding with the perspectives of users, in order to develop innovative ways to meet the mental health needs of under-served groups. PMID- 20003276 TI - Resistance exercise training lowers HbA1c more than aerobic training in adults with type 2 diabetes. AB - BACKGROUND: The aim of this study was to compare the effects of 10 weeks of resistance or treadmill exercises on glycemic indices levels prior to and immediately following exercise in adults with type 2 diabetes. RESEARCH DESIGN AND METHOD: Twenty inactive subjects (mean age 53.5 years) with type 2 diabetes enrolled in the study. Baseline HbA1c, blood glucose levels, heart rate, and blood pressure were measured for each subject prior to the initiation of the exercise program. Subsequently, subjects were matched to age, waist circumference and sex and assigned to either isocaloric resistance or treadmill exercise groups, which met 3 times per week for 10 weeks. RESULTS: Both groups showed a reduction in pre and post-exercise blood glucose and HbA1c values. There was no change in resting blood pressure or heart rate in either group during the course of the 10 week intervention. The group receiving resistance exercises showed significant differences in the daily pre-exercise plasma glucose readings between the beginning and end of the exercise protocol (p < 0.001). There were significant improvements in the mean HbA1c reading pre and post training in both groups (p < 0.001). However, the greater reduction was noted in the resistance exercise group, and at 10 weeks their HbA1c levels were significantly lower than the group that received treadmill exercises (p < 0.006). CONCLUSION: Ten weeks of resistance exercises were associated with a significantly better glycemic control in adults with type 2 diabetes compared to treadmill exercise. PMID- 20003277 TI - Early pregnancy peripheral blood gene expression and risk of preterm delivery: a nested case control study. AB - BACKGROUND: Preterm delivery (PTD) is a significant public health problem associated with greater risk of mortality and morbidity in infants and mothers. Pathophysiologic processes that may lead to PTD start early in pregnancy. We investigated early pregnancy peripheral blood global gene expression and PTD risk. METHODS: As part of a prospective study, ribonucleic acid was extracted from blood samples (collected at 16 weeks gestational age) from 14 women who had PTD (cases) and 16 women who delivered at term (controls). Gene expressions were measured using the GeneChip(R) Human Genome U133 Plus 2.0 Array. Student's T-test and fold change analysis were used to identify differentially expressed genes. We used hierarchical clustering and principle components analysis to characterize signature gene expression patterns among cases and controls. Pathway and promoter sequence analyses were used to investigate functions and functional relationships as well as regulatory regions of differentially expressed genes. RESULTS: A total of 209 genes, including potential candidate genes (e.g. PTGDS, prostaglandin D2 synthase 21 kDa), were differentially expressed. A set of these genes achieved accurate pre-diagnostic separation of cases and controls. These genes participate in functions related to immune system and inflammation, organ development, metabolism (lipid, carbohydrate and amino acid) and cell signaling. Binding sites of putative transcription factors such as EGR1 (early growth response 1), TFAP2A (transcription factor AP2A), Sp1 (specificity protein 1) and Sp3 (specificity protein 3) were over represented in promoter regions of differentially expressed genes. Real-time PCR confirmed microarray expression measurements of selected genes. CONCLUSIONS: PTD is associated with maternal early pregnancy peripheral blood gene expression changes. Maternal early pregnancy peripheral blood gene expression patterns may be useful for better understanding of PTD pathophysiology and PTD risk prediction. PMID- 20003278 TI - A combination of methotrexate and zoledronic acid prevents bone erosions and systemic bone mass loss in collagen induced arthritis. AB - INTRODUCTION: Osteoclasts play a key role in the pathogenesis of bone erosion and systemic bone mass loss during rheumatoid arthritis (RA). In this study, we aimed to determine the effect of methotrexate (MTX) and zoledronic acid (ZA), used alone or in combination, on osteoclast-mediated bone erosions and systemic bone mass loss in a rat model of collagen induced arthritis (CIA). We hypothesized that MTX and ZA could have an additive effect to prevent both bone erosion and systemic bone loss. METHODS: Arthritis was induced in 64 female Sprague-Dawley rats. After the clinical onset of CIA, rats were assigned to treatment with MTX (1 mg/kg/week), ZA (100 microg/kg twice weekly), both treatments at the same regimens, or vehicle. Arthritis score and paw thickness were recorded twice weekly. The rats were sacrificed on D28 and hind paws were removed for radiographic, histological and immunohistochemical analysis. The effects of treatments on osteoclastogenesis were determined by Tartrate resistant acid phosphatase (TRAP) staining. Micro-CT of the tibia was carried out for histomorphometric analysis. Bone mass density was evaluated by densitometry. RESULTS: MTX significantly decreased the severity of CIA, whereas ZA slightly exacerbated it. When these two drugs were used in combination, MTX prevented the pro-inflammatory effect of ZA. The combination of ZA with MTX was more effective than MTX alone for reducing structural joint damage with a dramatic decrease of osteoclasts' number in the eroded joints. However, MTX alone also significantly reduced the number of osteoclasts and the number of CD68+ mononuclear cells. ZA alone, or ZA with MTX, significantly increased the systemic bone mass density measured by densitometry and bone volume on histomorphometric analysis. CONCLUSIONS: A combination of MTX and ZA prevented both bone erosion and systemic bone loss in a rat model of arthritis. Both treatments independently decreased the number of osteoclasts in the eroded joint. However, while MTX probably acts mainly through a decrease of inflammation, ZA has a direct effect on osteoclasts, allowing a dramatic down-regulation of these cells in inflamed joints. These two different mechanisms of action provide support for the use of a combination of these two drugs to improve the prevention of structural joint damage in RA. PMID- 20003279 TI - Association of ADAM33 gene polymorphisms with COPD in a northeastern Chinese population. AB - BACKGROUND: Chronic obstructive pulmonary disease (COPD) is influenced by both environmental and genetic factors. ADAM33 (a disintegrin and metalloproteinase 33) has been one of the most exciting candidate genes for asthma since its first association with the disease in Caucasian populations. Recently, ADAM33 was shown to be associated with excessive decline of lung function and COPD. The aim of this study was to evaluate the potential relationship between polymorphisms of ADAM33 and COPD in a Han population in northeastern China. METHODS: A total of 312 COPD patients and a control group of 319 healthy volunteers were recruited for this study. Eight polymorphic loci (V4, T+1, T2, T1, S2, S1, Q-1, and F+1) of ADAM33 were selected for genotyping. Genotypes were determined by using the polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method. RESULTS: Statistically significant differences in the distributions of the T2G, T1G, S2C, and Q-1G alleles between patients and controls were observed (P < 0.001, odds ratio (OR) = 2.81, 95% confidence interval (CI) = 2.19-3.61; P < 0.001, OR = 2.60, 95% CI = 2.06-3.30; P = 0.03, OR = 1.31, 95% CI = 1.02-1.69; and P < 0.001, OR = 1.93, 95% CI = 1.50-2.50, respectively). Haplotype analysis showed that the frequencies of the CGGGGAGC, CGGGGAGT, CGGGCAGC, and CGGGGGGC haplotypes were significantly higher in the case group than in the control group (P = 0.0002, 0.0001, 0.0005, and 0.0074, respectively). In contrast, the haplotype CGAAGAGC was more common in the control group than in the case group (P < 0.0001). CONCLUSION: These preliminary results suggest an association between ADAM33 polymorphisms and COPD in a Chinese Han population. PMID- 20003280 TI - Reducing health risk in family members of patients with type 2 diabetes: views of first degree relatives. AB - BACKGROUND: Patients with type 2 diabetes can have an important role in discussing health risk within families. This study aimed to establish the acceptability to first degree relatives towards their relative with type 2 diabetes intervening as health promoters in their own families, using the Health Belief Model as a theoretical framework for evaluation. METHODS: Cross-sectional questionnaire design. Survey questionnaire for first degree relative (sibling or child) mailed to a random sample of patients with type 2 diabetes registered with an urban hospital diabetes clinic (n = 607 eligible patients). Patients were asked to pass on questionnaires to one to two first degree relatives. RESULTS: Questionnaires were returned from 257 families (42% response rate) with two responses provided by 107 families (a total of 364 questionnaires). The majority (94%) of first degree relatives of patients with type 2 diabetes would like to be informed about reducing their risk. Half (48%) of respondents reported being spoken to by a relative with type 2 diabetes about their risk of diabetes. Those spoken to were more likely to see themselves at risk of diabetes, to worry about developing diabetes and to view diabetes as a serious condition. CONCLUSIONS: A role for patients with type 2 diabetes in discussing health risk in their family appears to be acceptable to many relatives. Discussion of risk and interventions to reduce health risk with their relatives should be encouraged in patients with type 2 diabetes. PMID- 20003281 TI - An e-health driven laboratory information system to support HIV treatment in Peru: E-quity for laboratory personnel, health providers and people living with HIV. AB - BACKGROUND: Peru has a concentrated HIV epidemic with an estimated 76,000 people living with HIV (PLHIV). Access to highly active antiretroviral therapy (HAART) expanded between 2004-2006 and the Peruvian National Institute of Health was named by the Ministry of Health as the institution responsible for carrying out testing to monitor the effectiveness of HAART. However, a national public health laboratory information system did not exist. We describe the design and implementation of an e-health driven, web-based laboratory information system- NETLAB--to communicate laboratory results for monitoring HAART to laboratory personnel, health providers and PLHIV. METHODS: We carried out a needs assessment of the existing public health laboratory system, which included the generation and subsequent review of flowcharts of laboratory testing processes to generate better, more efficient streamlined processes, improving them and eliminating duplications. Next, we designed NETLAB as a modular system, integrating key security functions. The system was implemented and evaluated. RESULTS: The three main components of the NETLAB system, registration, reporting and education, began operating in early 2007. The number of PLHIV with recorded CD4 counts and viral loads increased by 1.5 times, to reach 18,907. Publication of test results with NETLAB took an average of 1 day, compared to a pre-NETLAB average of 60 days. NETLAB reached 2,037 users, including 944 PLHIV and 1,093 health providers, during its first year and a half. The percentage of overall PLHIV and health providers who were aware of NETLAB and had a NETLAB password has also increased substantially. CONCLUSION: NETLAB is an effective laboratory management tool since it is directly integrated into the national laboratory system and streamlined existing processes at the local, regional and national levels. The system also represents the best possible source of timely laboratory information for health providers and PLHIV, allowing patients to access their own results and other helpful information about their health, extending the scope of HIV treatment beyond the health facility and providing a model for other countries to follow. The NETLAB system now includes 100 diseases of public health importance for which the Peruvian National Institute of Health and the network of public health laboratories provide testing and results. PMID- 20003282 TI - Genome scan to assess the respective role of host-plant and environmental constraints on the adaptation of a widespread insect. AB - BACKGROUND: The evolutionary success of phytophagous insects could result from their adaptation to different host-plants. Alternatively, the diversification of widespread species might be driven by adaptation along environmental gradients. To disentangle the respective roles of host-plant versus abiotic environmental variables acting on the genome of an oligophagous insect, we performed a genome scan using 83 unlinked AFLP markers on larvae of the large pine weevil collected on two host-plants (pine and spruce) in four forestry regions across Europe. RESULTS: At this large geographic scale, the global genetic differentiation was low and there was no isolation by distance pattern, suggesting that migration is overwhelming genetic drift in this species. In this context, the widely used frequentist methods to detect outliers (e.g. Dfdist), which assume migration - drift equilibrium are not the most appropriate approach. The implementation of a recently developed Bayesian approach, conceived to detect outliers even in non equilibrium situations, consistently detected 9 out of 83 loci as outliers. Eight of these were validated as outliers by multiple logistic regressions: six correlated with environmental variables, one with host-plant and one with the interaction between environmental variables and host-plant. CONCLUSION: These results suggest a relatively greater importance of abiotic environmental variables, as opposed to factors linked with the host-plant, in shaping genetic differentiation across the genome in this species. Logistic regression allows the nature of factors involved in locus-specific selection to be precisely identified and represents another step forward in the process of identifying adaptive loci. PMID- 20003283 TI - An improved empirical bayes approach to estimating differential gene expression in microarray time-course data: BETR (Bayesian Estimation of Temporal Regulation). AB - BACKGROUND: Microarray gene expression time-course experiments provide the opportunity to observe the evolution of transcriptional programs that cells use to respond to internal and external stimuli. Most commonly used methods for identifying differentially expressed genes treat each time point as independent and ignore important correlations, including those within samples and between sampling times. Therefore they do not make full use of the information intrinsic to the data, leading to a loss of power. RESULTS: We present a flexible random effects model that takes such correlations into account, improving our ability to detect genes that have sustained differential expression over more than one time point. By modeling the joint distribution of the samples that have been profiled across all time points, we gain sensitivity compared to a marginal analysis that examines each time point in isolation. We assign each gene a probability of differential expression using an empirical Bayes approach that reduces the effective number of parameters to be estimated. CONCLUSIONS: Based on results from theory, simulated data, and application to the genomic data presented here, we show that BETR has increased power to detect subtle differential expression in time-series data. The open-source R package betr is available through Bioconductor. BETR has also been incorporated in the freely-available, open source MeV software tool available from http://www.tm4.org/mev.html. PMID- 20003284 TI - Back disorders and lumbar load in nursing staff in geriatric care: a comparison of home-based care and nursing homes. AB - BACKGROUND: Back pain is one of the most frequent complaints in the nursing profession. Thus, the 12-month prevalence of pain in the lumbar spine in nursing staff is as high as 76%. Only a few representative studies have assessed the prevalence rates of back pain and its risk factors among nursing staff in nursing homes in comparison to staff in home-based care facilities. The present study accordingly investigates the prevalence in the lumbar and cervical spine and determines the physical workload to lifting and caring in geriatric care. METHODS: 1390 health care workers in nursing homes and home care participated in this cross sectional survey. The nursing staff members were examined by occupational physicians according to the principals of the multistep diagnosis of musculoskeletal disorders. Occupational exposure to daily care activities with patient transfers was measured by a standardised questionnaire. The lumbar load was calculated with the Mainz-Dortmund dose model. Information on ergonomic conditions were recorded from the management of the nursing homes. Comparisons of all outcome variables were made between both care settings. RESULTS: Complete documentation, including the findings from the occupational physicians and the questionnaire, was available for 41%. Staff in nursing homes had more often positive orthopaedic findings than staff in home care. At the same time the values calculated for lumbar load were found to be significant higher in staff in nursing homes than in home-based care: 45% vs. 6% were above the reference value. Nursing homes were well equipped with technical lifting aids, though their provision with assistive advices is unsatisfactory. Situation in home care seems worse, especially as the staff often has to get by without assistance. CONCLUSIONS: Future interventions should focus on counteracting work-related lumbar load among staff in nursing homes. Equipment and training in handling of assistive devices should be improved especially for staff working in home care. PMID- 20003285 TI - Predictors of the first cardiovascular event in patients with systemic lupus erythematosus - a prospective cohort study. AB - INTRODUCTION: Cardiovascular disease (CVD) is a major cause of premature mortality among Systemic lupus erythematosus (SLE) patients. Many studies have measured and evaluated risk factors for premature subclinical atherosclerosis, but few studies are prospective and few have evaluated risk factors for hard endpoints, i.e. clinically important cardiovascular events (CVE). We investigated the impact of traditional and lupus associated risk factors for the first ever CVE in a longitudinal cohort of SLE patients. METHODS: A total of 182 SLE patients (mean age 43.9 years) selected to be free of CVE were included. Cardiovascular and autoimmune biomarkers were measured on samples collected after overnight fasting at baseline. Clinical information was collected at baseline and at follow up. End point was the first ever CVE (ischemic heart, cerebrovascular or peripheral vascular disease or death due to CVD). Impact of baseline characteristics/biomarkers on the risk of having a first CVE was evaluated with Cox regression. RESULTS: Follow up was 99.5% after a mean time of 8.3 years. Twenty-four patients (13%) had a first CVE. In age-adjusted Cox regression, any positive antiphospholipid antibody (aPL), elevated markers of endothelial activation (von Willebrand factor (vWf), soluble vascular cellular adhesion molecule-1 (sVCAM-1)) and fibrinogen predicted CVEs. Of SLE manifestations, arthritis, pleuritis and previous venous occlusion were positively associated with future CVEs while thrombocytopenia was negatively associated. Among traditional risk factors only age and smoking were significant predictors. In a multivariable Cox regression model age, any positive aPL, vWf and absence of thrombocytopenia were all predictors of the first CVE. CONCLUSIONS: In addition to age, positive aPL, biomarkers indicating increased endothelial cell activity/damage, and absence of thrombocytopenia were independent predictors of CVEs in this prospective study. Our results indicate that activation of the endothelium and the coagulation system are important features in SLE related CVD. Furthermore, we observed that the risk of CVEs seems to differ between subgroups of SLE patients. PMID- 20003286 TI - HER2 overexpression and amplification is present in a subset of ovarian mucinous carcinomas and can be targeted with trastuzumab therapy. AB - BACKGROUND: The response rate of ovarian mucinous carcinomas to paclitaxel/carboplatin is low, prompting interest in targeted molecular therapies. We investigated HER2 expression and amplification, and the potential for trastuzumab therapy in this histologic subtype of ovarian cancer. METHODS: HER2 status was tested in 33 mucinous carcinomas and 16 mucinous borderline ovarian tumors (BOT)). Five cases with documented recurrence and with tissue from the recurrence available for testing were analyzed to determine whether HER2 amplification status changed over time. Three prospectively identified recurrent mucinous ovarian carcinomas were assessed for HER2 amplification and patients received trastuzumab therapy with conventional chemotherapy. RESULTS: Amplification of HER2 was observed in 6/33 (18.2%) mucinous carcinomas and 3/16 (18.8%) BOT. HER2 amplification in primary mucinous carcinomas was not associated with an increased likelihood of recurrence. The prospectively identified recurrent mucinous carcinomas showed overexpression and amplification of HER2; one patient's tumor responded dramatically to trastuzumab in combination with conventional chemotherapy, while another patient experienced an isolated central nervous system recurrence after trastuzumab therapy. CONCLUSION: HER2 amplification is relatively common in ovarian mucinous carcinomas (6/33, 18.2%), although not of prognostic significance. Trastuzumab therapy is a treatment option for patients with mucinous carcinoma when the tumor has HER2 amplification and overexpression. PMID- 20003287 TI - Amygdala temporal dynamics: temperamental differences in the timing of amygdala response to familiar and novel faces. AB - BACKGROUND: Inhibited temperament - the predisposition to respond to new people, places or things with wariness or avoidance behaviors - is associated with increased risk for social anxiety disorder and major depression. Although the magnitude of the amygdala's response to novelty has been identified as a neural substrate of inhibited temperament, there may also be differences in temporal dynamics (latency, duration, and peak). We hypothesized that persons with inhibited temperament would have faster responses to novel relative to familiar neutral faces compared to persons with uninhibited temperament. We used event related functional magnetic resonance imaging to measure the temporal dynamics of the blood oxygen level dependent (BOLD) response to both novel and familiar neutral faces in participants with inhibited or uninhibited temperament. RESULTS: Inhibited participants had faster amygdala responses to novel compared with familiar faces, and both longer and greater amygdala response to all faces. There were no differences in peak response. CONCLUSION: Faster amygdala response to novelty may reflect a computational bias that leads to greater neophobic responses and represents a mechanism for the development of social anxiety. PMID- 20003288 TI - A malaria outbreak in Naxalbari, Darjeeling district, West Bengal, India, 2005: weaknesses in disease control, important risk factors. AB - An outbreak of malaria in Naxalbari, West Bengal, India, in 2005 was investigated to understand determinants and propose control measures. Malaria cases were slide confirmed. Methods included calculation of annual blood examination rates (ABER, number of slides examined/population), collection of water specimens from potential vector-breeding sites, sorting of villages in categories depending on the number of abandoned wells within two kilometers radius and review of the DDT spray coverage. Cases were compared with matched neighbourhood controls in terms of personal protection using matched odds ratios (MOR). 7,303 cases and 17 deaths were reported between April 2005 and March 2006 with a peak during October rains (Attack rate: 50 per 1,000, case fatality: 0.2%). The attack rate increased according to the number of abandoned wells within 2 kilometres radius (P < 0.0001, Chi-square for trend). Abandoned wells were Anopheles breeding sites. Compared with controls, cases were more likely to sleep outdoors (MOR: 3.8) and less likely to use of mosquito nets and repellents (MOR: 0.3 and 0.1, respectively). DDT spray coverage and ABER were 39% and 3.5%, below the recommended 85% and 10%, respectively. Overall, this outbreak resulted from weaknesses in malaria control measures and a combination of factors, including vector breeding, low implementation of personal protection and weak case detection. PMID- 20003289 TI - Refining gene signatures: a Bayesian approach. AB - BACKGROUND: In high density arrays, the identification of relevant genes for disease classification is complicated by not only the curse of dimensionality but also the highly correlated nature of the array data. In this paper, we are interested in the question of how many and which genes should be selected for a disease class prediction. Our work consists of a Bayesian supervised statistical learning approach to refine gene signatures with a regularization which penalizes for the correlation between the variables selected. RESULTS: Our simulation results show that we can most often recover the correct subset of genes that predict the class as compared to other methods, even when accuracy and subset size remain the same. On real microarray datasets, we show that our approach can refine gene signatures to obtain either the same or better predictive performance than other existing methods with a smaller number of genes. CONCLUSIONS: Our novel Bayesian approach includes a prior which penalizes highly correlated features in model selection and is able to extract key genes in the highly correlated context of microarray data. The methodology in the paper is described in the context of microarray data, but can be applied to any array data (such as micro RNA, for example) as a first step towards predictive modeling of cancer pathways. A user-friendly software implementation of the method is available. PMID- 20003290 TI - Self-inflicted penetrating eye injuries using a razor blade: case report. AB - BACKGROUND: A 23 year old white male with a history of social and behavioural problems attempted to blind himself chemically, with alcohol, and mechanically, with a razor blade. METHODS: Observational case report of a patient who self inflicted bilateral scleral lacerations with a razor blade, after losing his job. RESULTS: The patient sustained bilateral inferior scleral perforations, with hypotony and a right traumatic cataract. He received urgent surgical repair, and prophylactic antibiotics. There were no retinal breaks or detachments. He later underwent successful cataract surgery to the right eye. CONCLUSION: Self inflicted ocular injury may be possible in non-psychotic patients, as a situational response to a life event. Urgent repair can completely restore vision in some cases. Referral for psychiatric counseling is mandatory. PMID- 20003291 TI - Time dependence of changes of two cartilage layers in anterior cruciate ligament insertion after resection on chondrocyte apoptosis and decrease in glycosaminoglycan. AB - BACKGROUND: The purpose of this study is to clarify the differences in time dependent histological changes (chondrocyte apoptosis and glycosaminoglycan (GAG) layer thickness decrease) between uncalcified fibrocartilage (UF) and calcified fibrocartilage (CF) layers at the anterior cruciate ligament (ACL) insertion after ACL resection of rabbits. METHODS: Forty male Japanese white rabbits underwent ACL substance resection in the right knee (resection group) and same operation without resection in the left knee (sham group). Animals were sacrificed 1, 2, 4 and 6 weeks after surgery. RESULTS: In the UF layer, the apoptosis rate in the resection group was significantly higher than that in the sham group at 1 and 2 weeks. The GAG layer thicknesses of the UF layer in the resection group at 1, 2, 4 and 6 weeks were lower than those in the sham group. In the CF layer, the apoptosis rate in the resection group was significantly higher than that in the sham group at 2 and 4 weeks. The GAG layer thickness of the CF layer in the resection group was lower than that in the sham group only at 6 weeks. CONCLUSION: The increase in chondrocyte apoptosis rate preceded the decrease in GAG layer thickness in both layers. In the UF layer, the increase in chondrocyte apoptosis rate and the decrease in GAG layer thickness preceded those in the CF layer. Using a surviving ligament and minimizing a debridement of ACL remnant during ACL reconstruction may be important to maintain cartilage layers of ACL insertion. An injured ACL should be repaired before degenerative changes of the insertion occur. PMID- 20003292 TI - Effects of Cowpea mottle virus and Cucumber mosaic virus on six Soybean (Glycine max L.) cultivars. AB - The study was carried out to determine the comparative pathogenic response of six cultivars of soybean; TGx 1844-18E, TGx 1448-2E, TGx 1910-8F, TGx 1019-2EN, TGx 1910-8F and TGx 1876-4E to single and mixed infections with cowpea mottle virus and cucumber mosaic virus. The experiment was conducted in the screenhouse at the crop production pavilion, Faculty of Agriculture, University of Ilorin, Ilorin, Kwara state Nigeria. The results of the experiment revealed that all soybean cultivars were susceptible to single and mixed infection of the two viruses but to seemingly different extent. The single infection with cowpea mottle virus (CMeV), however, caused the most severe symptoms on the soybean cultivars. Cucumber mosaic virus (CMV) alone was not as severe as the CMeV. The mixed infection of CMeV and CMV did not cause higher severity than CMeV alone indicating that there was little or no synergistic effect between the two viruses on soybean. PMID- 20003293 TI - Effects of dietary phytoestrogens on plasma testosterone and triiodothyronine (T3) levels in male goat kids. AB - BACKGROUND: Exposure to xenoestrogens in humans and animals has gained increasing attention due to the effects of these compounds on reproduction. The present study was undertaken to investigate the influence of low-dose dietary phytoestrogen exposure, i.e. a mixture of genistein, daidzein, biochanin A and formononetin, on the establishment of testosterone production during puberty in male goat kids. METHODS: Goat kids at the age of 3 months received either a standard diet or a diet supplemented with phytoestrogens (3-4 mg/kg/day) for approximately 3 months. Plasma testosterone and total and free triiodothyronine (T3) concentrations were determined weekly. Testicular levels of testosterone and cAMP were measured at the end of the experiment. Repeated measurement analysis of variance using the MIXED procedure on the generated averages, according to the Statistical Analysis System program package (Release 6.12, 1996, SAS Institute Inc., Cary, NC, USA) was carried out. RESULTS: No significant difference in plasma testosterone concentration between the groups was detected during the first 7 weeks. However, at the age of 5 months (i.e. October 1, week 8) phytoestrogen-treated animals showed significantly higher testosterone concentrations than control animals (37.5 nmol/l vs 19.1 nmol/l). This elevation was preceded by a rise in plasma total T3 that occurred on September 17 (week 6). A slightly higher concentration of free T3 was detected in the phytoestrogen group at the same time point, but it was not until October 8 and 15 (week 9 and 10) that a significant difference was found between the groups. At the termination of the experiment, testicular cAMP levels were significantly lower in goats fed a phytoestrogen-supplemented diet. Phytoestrogen-fed animals also had lower plasma and testicular testosterone concentrations, but these differences were not statistically significant. CONCLUSION: Our findings suggest that phytoestrogens can stimulate testosterone synthesis during puberty in male goats by increasing the secretion of T3; a hormone known to stimulate Leydig cell steroidogenesis. It is possible that feedback signalling underlies the tendency towards decreased steroid production at the end of the experiment. PMID- 20003294 TI - Trends in healthcare utilization among older Americans with colorectal cancer: a retrospective database analysis. AB - BACKGROUND: Analyses of utilization trends (cost drivers) allow us to understand changes in colorectal cancer (CRC) costs over time, better predict future costs, identify changes in the use of specific types of care (eg, hospice), and provide inputs for cost-effectiveness models. This retrospective cohort study evaluated healthcare resource use among US Medicare beneficiaries diagnosed with CRC between 1992 and 2002. METHODS: Cohorts included patients aged 66+ newly diagnosed with adenocarcinoma of the colon (n = 52,371) or rectum (n = 18,619) between 1992 and 2002 and matched patients from the general Medicare population, followed until death or December 31, 2005. Demographic and clinical characteristics were evaluated by cancer subsite. Resource use, including the percentage that used each type of resource, number of hospitalizations, and number of hospital and skilled nursing facility days, was evaluated by stage and subsite. The number of office, outpatient, and inpatient visits per person-year was calculated for each cohort, and was described by year of service, subsite, and treatment phase. Hospice use rates in the last year of life were calculated by year of service, stage, and subsite for CRC patients who died of CRC. RESULTS: CRC patients (mean age: 77.3 years; 44.9% male) used more resources than controls in every category (P < .001), with the largest differences seen in hospital days and home health use. Most resource use (except hospice) remained relatively steady over time. The initial phase was the most resource intense in terms of office and outpatient visits. Hospice use among patients who died of CRC increased from 20.0% in 1992 to 70.5% in 2004, and age-related differences appear to have evened out in later years. CONCLUSION: Use of hospice care among CRC decedents increased substantially over the study period, while other resource use remained generally steady. Our findings may be useful for understanding CRC cost drivers, tracking trends, and forecasting resource needs for CRC patients in the future. PMID- 20003295 TI - The effects of HIF-1alpha on gene expression profiles of NCI-H446 human small cell lung cancer cells. AB - BACKGROUND: Gene targeted therapy refers to any therapy focused on one of the many biological features of the tumor. Such features are mediated by specific genes that are involved in tumor metastasis, recurrence, poor response to chemotherapy and others. Hypoxia is an important pathognomonic feature of many malignant tumors including SCLC (small cell lung cancer). HIF-1alpha, which is induced by hypoxia, is the most important regulatory factor of many specific genes that can influence the biological features of tumors. METHODS: In this study, we tried to elucidate the changes in gene expression profiles of SCLC NCI H446 cells mediated by HIF-1alpha. According to different treatments of cells, three experimental pairwise comparisons were designed: hypoxia group vs. control group, Ad5-HIF-1alpha group vs. Ad5 group, and Ad5-siHIF-1 alpha group Vs Ad5 group. RESULTS: Results from the analysis of gene expression profiles indicated that there were 65 genes upregulated and 28 genes downregulated more than two fold in all three experimental pairwise comparisons. These genes were involved in transport, signal-transduction, cell adhesion/motility, growth factor/cytokines, transcription, inflammatory response, metabolic process, in addition to others. SOCS1, IGFBP5, IL-6 and STAT3 were also upregulated at protein level. SOCS1 could significantly induce apoptosis and suppress growth of NCI-H446 cells but HIF 1alpha could induce growth and suppress apoptosis. CONCLUSIONS: Through this research, we are trying to find novel functional genes that are mediated by HIF 1alpha and provide the theoretical basis for new therapeutic targets. HIF-1 alpha maybe upregulate the expression of SOCS1 through mediation of STAT3 and IL-6. In addition, SOCS1 could significantly induce apoptosis and suppress growth of NCI H446 cells. This was contrary to HIF-1alpha and it indicated that there might be an antagonism effect between HIF-1alpha and SOCS1 on regulating growth and apoptosis of NCI-H446 cells. PMID- 20003296 TI - Proteome analysis of Aspergillus niger: lactate added in starch-containing medium can increase production of the mycotoxin fumonisin B2 by modifying acetyl-CoA metabolism. AB - BACKGROUND: Aspergillus niger is a filamentous fungus found in the environment, on foods and feeds and is used as host for production of organic acids, enzymes and proteins. The mycotoxin fumonisin B2 was recently found to be produced by A. niger and hence very little is known about production and regulation of this metabolite. Proteome analysis was used with the purpose to reveal how fumonisin B2 production by A. niger is influenced by starch and lactate in the medium. RESULTS: Fumonisin B2 production by A. niger was significantly increased when lactate and starch were combined in the medium. Production of a few other A. niger secondary metabolites was affected similarly by lactate and starch (fumonisin B4, orlandin, desmethylkotanin and pyranonigrin A), while production of others was not (ochratoxin A, ochratoxin alpha, malformin A, malformin C, kotanin, aurasperone B and tensidol B). The proteome of A. niger was clearly different during growth on media containing 3% starch, 3% starch + 3% lactate or 3% lactate. The identity of 59 spots was obtained, mainly those showing higher or lower expression levels on medium with starch and lactate. Many of them were enzymes in primary metabolism and other processes that affect the intracellular level of acetyl-CoA or NADPH. This included enzymes in the pentose phosphate pathway, pyruvate metabolism, the tricarboxylic acid cycle, ammonium assimilation, fatty acid biosynthesis and oxidative stress protection. CONCLUSIONS: Lactate added in a medium containing nitrate and starch can increase fumonisin B2 production by A. niger as well as production of some other secondary metabolites. Changes in the balance of intracellular metabolites towards a higher level of carbon passing through acetyl-CoA and a high capacity to regenerate NADPH during growth on medium with starch and lactate were found to be the likely cause of this effect. The results lead to the hypothesis that fumonisin production by A. niger is regulated by acetyl-CoA. PMID- 20003297 TI - Expansion of tandem repeats in sea anemone Nematostella vectensis proteome: A source for gene novelty? AB - BACKGROUND: The complete proteome of the starlet sea anemone, Nematostella vectensis, provides insights into gene invention dating back to the Cnidarian Bilaterian ancestor. With the addition of the complete proteomes of Hydra magnipapillata and Monosiga brevicollis, the investigation of proteins having unique features in early metazoan life has become practical. We focused on the properties and the evolutionary trends of tandem repeat (TR) sequences in Cnidaria proteomes. RESULTS: We found that 11-16% of N. vectensis proteins contain tandem repeats. Most TRs cover 150 amino acid segments that are comprised of basic units of 5-20 amino acids. In total, the N. Vectensis proteome has about 3300 unique TR-units, but only a small fraction of them are shared with H. magnipapillata, M. brevicollis, or mammalian proteomes. The overall abundance of these TRs stands out relative to that of 14 proteomes representing the diversity among eukaryotes and within the metazoan world. TR-units are characterized by a unique composition of amino acids, with cysteine and histidine being over represented. Structurally, most TR-segments are associated with coiled and disordered regions. Interestingly, 80% of the TR-segments can be read in more than one open reading frame. For over 100 of them, translation of the alternative frames would result in long proteins. Most domain families that are characterized as repeats in eukaryotes are found in the TR-proteomes from Nematostella and Hydra. CONCLUSIONS: While most TR-proteins have originated from prediction tools and are still awaiting experimental validations, supportive evidence exists for hundreds of TR-units in Nematostella. The existence of TR-proteins in early metazoan life may have served as a robust mode for novel genes with previously overlooked structural and functional characteristics. PMID- 20003298 TI - Evaluation of prescribing patterns in a German network of CAM physicians for the treatment of patients with hypertension: a prospective observational study. AB - BACKGROUND: The management of hypertension is a key challenge in modern health systems. This study aimed to investigate hypertension treatment strategies among physicians specialized in complementary and alternative medicine (CAM) in Germany by analysing prescribing patterns and comparing these to the current treatment guidelines issued by the German Hypertension Society. METHODS: In this prospective, multicentre observational study, which included 25 primary care physicians specialized in CAM treatment, prescriptions and diagnoses were analysed for each consecutive hypertensive patient using routine electronic data. Data analysis was performed using univariate statistical tests (Chi square test, Cochran-Armitage trend test). Multiple logistic regression was used to determine factors associated with antihypertensive medication. RESULTS: In the year 2005, 1320 patients with 3278 prescriptions were included (mean age = 64.2 years (SD = 14.5), 63.5% women). Most patients were treated with conventional antihypertensive monotherapies (n = 838, 63.5%). Beta-blockers were the most commonly prescribed monotherapy (30.7%), followed by ACE inhibitors (24.0%). Combination treatment usually consisted of two antihypertensive drugs administered either as separate agents or as a coformulation. The most common combination was a diuretic plus an ACE inhibitor (31.2% of dual therapies). Patient gender, age, and comorbidities significantly influenced which treatment was prescribed. 187 patients (14.2%) received one or more CAM remedies, most of which were administered in addition to classic monotherapies (n = 104). Men (OR = 0.66; 95% CI: 0.54-0.80) and patients with diabetes (OR = 0.55; 95% CI: 0.42 0.0.73), hypercholesterolaemia (OR = 0.59; 95% CI: 0.47-0.75), obesity (OR = 0.74; 95% CI: 0.57-0.97), stroke (OR = 0.54; 95% CI: 0.40-0.74), or prior myocardial infarction (OR = 0.37; 95% CI: 0.17-0.81) were less likely to receive CAM treatment. CONCLUSIONS: The large majority of antihypertensive treatments prescribed by CAM physicians in the present study complied with the current German Hypertension Society treatment guidelines. Deviations from the guidelines were observed in one of every seven patients receiving some form of CAM treatment. PMID- 20003299 TI - Protocol for the Osteoporosis Choice trial. A pilot randomized trial of a decision aid in primary care practice. AB - BACKGROUND: Bisphosphonates can reduce fracture risk in patients with osteoporosis, but many at-risk patients do not start or adhere to these medications. The aims of this study are to: (1) preliminarily evaluate the effect of an individualized 10-year osteoporotic fracture risk calculator and decision aid (OSTEOPOROSIS CHOICE) for postmenopausal women at risk for osteoporotic fractures; and (2) assess the feasibility and validity (i.e., absence of contamination) of patient-level randomization (vs. cluster randomization) in pilot trials of decision aid efficacy. METHODS/DESIGN: This is a protocol for a parallel, 2-arm, randomized trial to compare an intervention group receiving OSTEOPOROSIS CHOICE to a control group receiving usual primary care. Postmenopausal women with bone mineral density T-scores of <-1.0, not receiving bisphosphonate therapy, and receiving care at participating primary care practices in and around Rochester, Minnesota, USA will be eligible to participate in the trial. We will measure the effect of OSTEOPOROSIS CHOICE on five outcomes: (a) patient knowledge regarding osteoporosis risk factors and treatment; (b) quality of the decision-making process for both the patient and clinician; (c) patient and clinician acceptability and satisfaction with the decision aid; (d) rate of bisphosphonate use and adherence, and (e) trial processes (e.g., ability to recruit participants, collect patient outcomes). To capture these outcomes, we will use patient and clinician surveys following each visit and video recordings of the clinical encounters. These video recordings will also allow us to determine the extent to which clinicians previously exposed to the decision aid were able to recreate elements of the decision aid with control patients (i.e., contamination). Pharmacy prescription profiles and follow-up phone interviews will assess medication start and adherence at 6 months. DISCUSSION: This pilot trial will provide evidence of feasibility, validity of patient randomization, and preliminary efficacy of a novel approach--decision aids--to improving medication adherence for postmenopausal women at risk of osteoporotic fractures. The results will inform the design of a larger trial that could provide more precise estimates of the efficacy of the decision aid. TRIAL REGISTRATION: Clinical Trials.gov Identifier: NCT00578981. PMID- 20003300 TI - The effectiveness of health coaching, home blood pressure monitoring, and home titration in controlling hypertension among low-income patients: protocol for a randomized controlled trial. AB - BACKGROUND: Despite the many antihypertensive medications available, two-thirds of patients with hypertension do not achieve blood pressure control. This is thought to be due to a combination of poor patient education, poor medication adherence, and "clinical inertia." The present trial evaluates an intervention consisting of health coaching, home blood pressure monitoring, and home medication titration as a method to address these three causes of poor hypertension control. METHODS/DESIGN: The randomized controlled trial will include 300 patients with poorly controlled hypertension. Participants will be recruited from a primary care clinic in a teaching hospital that primarily serves low-income populations.An intervention group of 150 participants will receive health coaching, home blood pressure monitoring, and home-titration of antihypertensive medications during 6 months. The control group (n=150) will receive health coaching plus home blood pressure monitoring for the same duration. A passive control group will receive usual care. Blood pressure measurements will take place at baseline, and after 6 and 12 months. The primary outcome will be change in systolic blood pressure after 6 and 12 months. Secondary outcomes measured will be change in diastolic blood pressure, adverse events, and patient and provider satisfaction. DISCUSSION: The present study is designed to assess whether the 3-pronged approach of health coaching, home blood pressure monitoring, and home medication titration can successfully improve blood pressure, and if so, whether this effect persists beyond the period of the intervention. TRIAL REGISTRATION: ClinicalTrials.gov identifier: NCT01013857. PMID- 20003301 TI - Towards accurate imputation of quantitative genetic interactions. AB - Recent technological breakthroughs have enabled high-throughput quantitative measurements of hundreds of thousands of genetic interactions among hundreds of genes in Saccharomyces cerevisiae. However, these assays often fail to measure the genetic interactions among up to 40% of the studied gene pairs. Here we present a novel method, which combines genetic interaction data together with diverse genomic data, to quantitatively impute these missing interactions. We also present data on almost 190,000 novel interactions. PMID- 20003302 TI - Demographic costs of inbreeding revealed by sex-specific genetic rescue effects. AB - BACKGROUND: Inbreeding can slow population growth and elevate extinction risk. A small number of unrelated immigrants to an inbred population can substantially reduce inbreeding and improve fitness, but little attention has been paid to the sex-specific effects of immigrants on such "genetic rescue". We conducted two subsequent experiments to investigate demographic consequences of inbreeding and genetic rescue in guppies. RESULTS: Populations established from pairs of full siblings that were descended either from two generations of full-sibling inbreeding or unrelated outbred guppies did not grow at different rates initially, but when the first generation offspring started breeding, outbred founded populations grew more slowly than inbred-founded populations. In a second experiment, adding two outbred males to the inbred populations resulted in significantly faster population growth than in control populations where no immigrants were added. Adding females resulted in growth at a rate intermediate to the control and male-immigrant treatments. CONCLUSION: The slower growth of the outbred-founded than inbred-founded populations is the opposite of what would be expected under inbreeding depression unless many deleterious recessive alleles had already been selectively purged in the inbreeding that preceded the start of the experiment, and that significant inbreeding depression occurred when the first generation offspring in outbred-founded populations started to inbreed. The second experiment revealed strong inbreeding depression in the inbred founded populations, despite the apparent lack thereof in these populations earlier on. Moreover, the fact that the addition of male immigrants resulted in the highest levels of population growth suggests that sex-specific genetic rescue may occur in promiscuous species, with male rescue resulting in higher levels of outbreeding than female rescue. PMID- 20003303 TI - Insight into microRNA regulation by analyzing the characteristics of their targets in humans. AB - BACKGROUND: microRNAs (miRNAs) are believed to regulate their targets through posttranscriptional gene regulation and have the potential to silence gene expression via multiple mechanisms. Despite previous advances on miRNA regulation of gene expression, little has been investigated from a genome scale. RESULTS: To gain new insight into miRNA regulation in humans, we used large scale data and carried out a series of studies to compare various features of miRNA target genes to that of non-miRNA target genes. We observed significant differences between miRNA and non-miRNA target genes for a number of characteristics, including higher and broader mRNA expression, faster mRNA decay rate, longer protein half life, and longer gene structures. Based on these features and by analyzing their relationships we found that miRNA target genes, other than having miRNA repression, were most likely under more complex regulation than non-miRNA target genes, which was evidenced by their higher and broader gene expression but longer gene structures. Our results of higher and broader gene expression but fast mRNA decay rates also provide evidence that miRNA dampening of the output of preexisting transcripts facilitates a more rapid and robust transition to new expression programs. This could be achieved by enhancing mRNA degradation through an additive effect from multiple miRNA targeting. CONCLUSION: Genome-scale analysis on the nature of miRNA target genes has revealed a general mechanism for miRNA regulation of human gene expression. The results of this study also indicate that miRNA target genes, other than having miRNA repression, are under more complex gene regulation than non-miRNA target genes. These findings provide novel insight into miRNA regulation of human gene expression. PMID- 20003304 TI - A survey on knowledge and self-reported formula handling practices of parents and child care workers in Palermo, Italy. AB - BACKGROUND: Powdered infant formula (PIF) is not a sterile product, but this information appears to be poorly diffused among child caregivers. Parents and child care workers may behave in an unsafe manner when handling PIF. METHODS: This study involved parents and child care workers in the 24 municipal child care centres of Palermo. Knowledge and self-reported practices about PIF handling were investigated by a structured questionnaire. A Likert scale was used to measure the strength of the respondent's feelings. Association of knowledge and self reported practices with demographic variables was also evaluated. RESULTS: 42.4% of parents and 71.0% of child care workers filled in the questionnaire. Significant differences were found between parents and child care workers for age and education. 73.2% of parents and 84.4% of child care workers were confident in sterility of PIF. Generally, adherence to safe procedures when reconstituting and handling PIF was more frequently reported by child care workers who, according to the existing legislation, are regularly subjected to a periodic training on food safety principles and practices. Age and education significantly influenced the answers to the questionnaire of both parents and child care workers. CONCLUSION: The results of the study reveal that parents and child care workers are generally unaware that powdered formulas may contain viable microorganisms. However, child care workers consistently chose safer options than parents when answering the questions about adherence to hygienic practices.At present it seems unfeasible to produce sterile PIF, but the risk of growth of hazardous organisms in formula at the time of administration should be minimized by promoting safer behaviours among caregivers to infants in both institutional settings and home. PMID- 20003305 TI - A recent duplication revisited: phylogenetic analysis reveals an ancestral duplication highly-conserved throughout the Oryza genus and beyond. AB - BACKGROUND: The role of gene duplication in the structural and functional evolution of genomes has been well documented. Analysis of complete rice (Oryza sativa) genome sequences suggested an ancient whole genome duplication, common to all the grasses, some 50-70 million years ago and a more conserved segmental duplication between the distal regions of the short arms of chromosomes 11 and 12, whose evolutionary history is controversial. RESULTS: We have carried out a comparative analysis of this duplication within the wild species of the genus Oryza, using a phylogenetic approach to specify its origin and evolutionary dynamics. Paralogous pairs were isolated for nine genes selected throughout the region in all Oryza genome types, as well as in two outgroup species, Leersia perrieri and Potamophila parviflora. All Oryza species display the same global evolutionary dynamics but some lineage-specific features appear towards the proximal end of the duplicated region. The same level of conservation is observed between the redundant copies of the tetraploid species Oryza minuta. The presence of orthologous duplicated blocks in the genome of the more distantly-related species, Brachypodium distachyon, strongly suggests that this duplication between chromosomes 11 and 12 was formed as part of the whole genome duplication common to all Poaceae. CONCLUSION: Our observations suggest that recurrent but heterogeneous concerted evolution throughout the Oryza genus and in related species has led specifically to the extremely high sequence conservation occurring in this region of more than 2 Mbp. PMID- 20003306 TI - Dietary intake, lung function and airway inflammation in Mexico City school children exposed to air pollutants. AB - INTRODUCTION: Air pollutant exposure has been associated with an increase in inflammatory markers and a decline in lung function in asthmatic children. Several studies suggest that dietary intake of fruits and vegetables might modify the adverse effect of air pollutants. METHODS: A total of 158 asthmatic children recruited at the Children's Hospital of Mexico and 50 non-asthmatic children were followed for 22 weeks. Pulmonary function was measured and nasal lavage collected and analyzed every 2 weeks. Dietary intake was evaluated using a 108-item food frequency questionnaire and a fruit and vegetable index (FVI) and a Mediterranean diet index (MDI) were constructed. The impact of these indices on lung function and interleukin-8 (IL-8) and their interaction with air pollutants were determined using mixed regression models with random intercept and random slope. RESULTS: FVI was inversely related to IL-8 levels in nasal lavage (p < 0.02) with a significant inverse trend (test for trend p < 0.001), MDI was positively related to lung function (p < 0.05), and children in the highest category of MDI had a higher FEV1 (test for trend p < 0.12) and FVC (test for trend p < 0.06) than children in the lowest category. A significant interaction was observed between FVI and ozone for FEV1 and FVC as was with MDI and ozone for FVC. No effect of diet was observed among healthy children. CONCLUSION: Our results suggest that fruit and vegetable intake and close adherence to the Mediterranean diet have a beneficial effect on inflammatory response and lung function in asthmatic children living in Mexico City. PMID- 20003307 TI - Deciphering the transcriptional circuitry of microRNA genes expressed during human monocytic differentiation. AB - BACKGROUND: Macrophages are immune cells involved in various biological processes including host defence, homeostasis, differentiation, and organogenesis. Disruption of macrophage biology has been linked to increased pathogen infection, inflammation and malignant diseases. Differential gene expression observed in monocytic differentiation is primarily regulated by interacting transcription factors (TFs). Current research suggests that microRNAs (miRNAs) degrade and repress translation of mRNA, but also may target genes involved in differentiation. We focus on getting insights into the transcriptional circuitry regulating miRNA genes expressed during monocytic differentiation. RESULTS: We computationally analysed the transcriptional circuitry of miRNA genes during monocytic differentiation using in vitro time-course expression data for TFs and miRNAs. A set of TF-->miRNA associations was derived from predicted TF binding sites in promoter regions of miRNA genes. Time-lagged expression correlation analysis was utilised to evaluate the TF-->miRNA associations. Our analysis identified 12 TFs that potentially play a central role in regulating miRNAs throughout the differentiation process. Six of these 12 TFs (ATF2, E2F3, HOXA4, NFE2L1, SP3, and YY1) have not previously been described to be important for monocytic differentiation. The remaining six TFs are CEBPB, CREB1, ELK1, NFE2L2, RUNX1, and USF2. For several miRNAs (miR-21, miR-155, miR-424, and miR-17-92), we show how their inferred transcriptional regulation impacts monocytic differentiation. CONCLUSIONS: The study demonstrates that miRNAs and their transcriptional regulatory control are integral molecular mechanisms during differentiation. Furthermore, it is the first study to decipher on a large-scale, how miRNAs are controlled by TFs during human monocytic differentiation. Subsequently, we have identified 12 candidate key controllers of miRNAs during this differentiation process. PMID- 20003308 TI - Increased immunogenicity of surviving tumor cells enables cooperation between liposomal doxorubicin and IL-18. AB - BACKGROUND: Liposomal doxorubicin (Doxil) is a cytotoxic chemotherapy drug with a favorable hematologic toxicity profile. Its active drug, doxorubicin, has interesting immunomodulatory properties. Here, the effects of Doxil on surviving tumor cell immunophenotype were investigated. METHODS: Using ID8 murine ovarian cancer cells, the immunomodulatory effects of Doxil were studied by measuring its impact on ovarian cancer cell expression of MHC class-I and Fas, and susceptibility to immune attack in vitro. To evaluate the ability of Doxil to cooperate with cancer immunotherapy, the interaction between Doxil and Interleukin 18 (IL-18), a pleiotropic immunostimulatory cytokine, was investigated in vivo in mice bearing ID8-Vegf tumors. RESULTS: While Doxil killed ID8 tumor cells in a dose-dependent manner, tumor cells escaping Doxil-induced apoptosis upregulated surface expression of MHC-I and Fas, and were sensitized to CTL killing and Fas-mediated death in vitro. We therefore tested the hypothesis that the combination of immunotherapy with Doxil provides positive interactions. Combination IL-18 and Doxil significantly suppressed tumor growth compared with either monotherapy in vivo and uniquely resulted in complete tumor regression and long term antitumor protection in a significant proportion of mice. CONCLUSION: These data demonstrate that Doxil favorably changes the immunophenotype of a large fraction of the tumor that escapes direct killing thus creating an opportunity to expand tumor killing by immunotherapy, which can be capitalized through addition of IL-18 in vivo. PMID- 20003310 TI - Social support during childbirth as a catalyst for early breastfeeding initiation for first-time Nigerian mothers. AB - BACKGROUND: Initiation of breastfeeding can be difficult in a busy maternity centre with inadequate manpower and social support. This study aims to explore the role of psychosocial support offered by companions on breastfeeding initiation among first-time mothers. METHODS: This is a secondary data analysis of a randomised controlled trial conducted among women attending the antenatal clinic of the University College Hospital, Ibadan, Nigeria in 2007. Those in the experimental group were asked to bring someone of their choice to the labour room to act as a companion; the comparison group received standard care. The results of 209 HIV negative women who had vaginal births were analysed. The main outcome measure was time to initiation of breastfeeding after childbirth. RESULTS: Of the total, 94 had companions during labour while 115 did not have a companion. The median time to breastfeeding initiation was significantly shorter in those with companions compared to controls (16 vs. 54 minutes; p < 0.01). The cumulative survival analysis indicated that all in the treatment group had initiated breastfeeding by 26 minutes, while among the control group none had commenced at 30 minutes post-delivery with some as late as 12 hours. After Cox regression analysis was used to adjust for possible confounders, the outcome still showed a significant hazard ratio of 207.8 (95%CI 49.2, 878.0; p < 0.01) among women who were supported by a companion. CONCLUSION: Use of companions during labour is associated with earlier time to breastfeeding initiation among first-time mothers in Nigeria. TRIAL REGISTRATION: Australian New Zealand Clinical Trials Registry: ACTRN12609000994280. PMID- 20003309 TI - Differential regulation of immune responses and macrophage/neuron interactions in the dorsal root ganglion in young and adult rats following nerve injury. AB - BACKGROUND: Neuropathic pain is an apparently spontaneous experience triggered by abnormal physiology of the peripheral or central nervous system, which evolves with time. Neuropathic pain arising from peripheral nerve injury is characterized by a combination of spontaneous pain, hyperalgesia and allodynia. There is no evidence of this type of pain in human infants or rat pups; brachial plexus avulsion, which causes intense neuropathic pain in adults, is not painful when the injury is sustained at birth. Since infants are capable of nociception from before birth and display both acute and chronic inflammatory pain behaviour from an early neonatal age, it appears that the mechanisms underlying neuropathic pain are differentially regulated over a prolonged postnatal period. RESULTS: We have performed a microarray analysis of the rat L4/L5 dorsal root ganglia (DRG), 7 days post spared nerve injury, a model of neuropathic pain. Genes that are regulated in adult rats displaying neuropathic behaviour were compared to those regulated in young rats (10 days old) that did not show the same neuropathic behaviour. The results show a set of genes, differentially regulated in the adult DRG, that are principally involved in immune system modulation. A functional consequence of this different immune response to injury is that resident macrophages cluster around the large A sensory neuron bodies in the adult DRG seven days post injury, whereas the macrophages in young DRG remain scattered evenly throughout the ganglion, as in controls. CONCLUSIONS: The results show, for the first time, a major difference in the neuroimmune response to nerve injury in the dorsal root ganglion of young and adult rats. Differential analysis reveals a new set of immune related genes in the ganglia, that are differentially regulated in adult neuropathic pain, and that are consistent with the selective activation of macrophages around adult, but not young large A sensory neurons post injury. These differences may contribute to the reduced incidence of neuropathic pain in infants. PMID- 20003311 TI - Combining a leadership course and multi-source feedback has no effect on leadership skills of leaders in postgraduate medical education. An intervention study with a control group. AB - BACKGROUND: Leadership courses and multi-source feedback are widely used developmental tools for leaders in health care. On this background we aimed to study the additional effect of a leadership course following a multi-source feedback procedure compared to multi-source feedback alone especially regarding development of leadership skills over time. METHODS: Study participants were consultants responsible for postgraduate medical education at clinical departments. STUDY DESIGN: pre-post measures with an intervention and control group. The intervention was participation in a seven-day leadership course. Scores of multi-source feedback from the consultants responsible for education and respondents (heads of department, consultants and doctors in specialist training) were collected before and one year after the intervention and analysed using Mann-Whitney's U-test and Multivariate analysis of variances. RESULTS: There were no differences in multi-source feedback scores at one year follow up compared to baseline measurements, either in the intervention or in the control group (p = 0.149). CONCLUSION: The study indicates that a leadership course following a MSF procedure compared to MSF alone does not improve leadership skills of consultants responsible for education in clinical departments. Developing leadership skills takes time and the time frame of one year might have been too short to show improvement in leadership skills of consultants responsible for education. Further studies are needed to investigate if other combination of initiatives to develop leadership might have more impact in the clinical setting. PMID- 20003312 TI - An efficient algorithm for the stochastic simulation of the hybridization of DNA to microarrays. AB - BACKGROUND: Although oligonucleotide microarray technology is ubiquitous in genomic research, reproducibility and standardization of expression measurements still concern many researchers. Cross-hybridization between microarray probes and non-target ssDNA has been implicated as a primary factor in sensitivity and selectivity loss. Since hybridization is a chemical process, it may be modeled at a population-level using a combination of material balance equations and thermodynamics. However, the hybridization reaction network may be exceptionally large for commercial arrays, which often possess at least one reporter per transcript. Quantification of the kinetics and equilibrium of exceptionally large chemical systems of this type is numerically infeasible with customary approaches. RESULTS: In this paper, we present a robust and computationally efficient algorithm for the simulation of hybridization processes underlying microarray assays. Our method may be utilized to identify the extent to which nucleic acid targets (e.g. cDNA) will cross-hybridize with probes, and by extension, characterize probe robustnessusing the information specified by MAGE TAB. Using this algorithm, we characterize cross-hybridization in a modified commercial microarray assay. CONCLUSIONS: By integrating stochastic simulation with thermodynamic prediction tools for DNA hybridization, one may robustly and rapidly characterize of the selectivity of a proposed microarray design at the probe and "system" levels. Our code is available at http://www.laurenzi.net. PMID- 20003313 TI - Mutations in the coding regions of the hepatocyte nuclear factor 4 alpha in Iranian families with maturity onset diabetes of the young. AB - Hepatocyte nuclear factor 4alpha (HNF4alpha) is a nuclear receptor involved in glucose homeostasis and is required for normal beta cell function. Mutations in the HNF4alpha gene are associated with maturity onset diabetes of the young type 1 (MODY1). The aim of the present study was to determine the prevalence and nature of mutations in HNF4alpha gene in Iranian patients with a clinical diagnosis of MODY and their family members. Twelve families including 30 patients with clinically MODY diagnosis and 21 members of their family were examined using PCR-RFLP method and in case of mutation confirmed by sequencing techniques. Fifty age and sex matched subjects with normal fasting blood sugar (FBS) and Glucose tolerance test (GTT) were constituted the control group and investigated in the similar pattern. Single mutation of V255M in the HNF4alpha gene was detected. This known mutation was found in 8 of 30 patients and 3 of 21 individuals in relatives. Fifty healthy control subjects did not show any mutation. Here, it is indicated that the prevalence of HNF4alpha mutation among Iranian patients with clinical MODY is considerable. This mutation was present in 26.6% of our patients, but nothing was found in control group. In the family members, 3 subjects with the age of or=18 on entry to study; and serum 25-hydroxyvitamin D levels <75 nmol/L. Patients will be randomised 1:1 to receive either oral cholecalciferol 2000IU/day or placebo for 6 months. The primary outcome will be an improvement in insulin sensitivity, measured by hyperinsulinaemic euglycaemic clamp. Secondary outcome measures will include serum parathyroid hormone, cytokines (Interleukin-1beta, Interleukin-6, Tumour Necrosis Factor alpha), adiponectin (total and High Molecular Weight), osteocalcin (carboxylated and under-carboxylated), peripheral blood mononuclear cell Nuclear Factor Kappa-B p65 binding activity, brachial artery reactivity, aortic pulse wave velocity and waveform analysis, and indirect calorimetry. All outcome measures will be performed at baseline and end of study. DISCUSSION: To date, no randomised controlled trial has been performed in pre-dialysis CKD patients to study the correlation between vitamin D status with supplementation, insulin resistance and markers of adverse cardiovascular risk. We remain hopeful that cholecalciferol may be a safe intervention, with health benefits beyond those related to bone-mineral homeostasis. TRIAL REGISTRATION: Australian and New Zealand Clinical Trials Registry ACTRN12609000246280. PMID- 20003317 TI - The neuroprotective properties of palmitoylethanolamine against oxidative stress in a neuronal cell line. AB - BACKGROUND: N-acylethanolamines (NAEs) are lipids upregulated in response to cell and tissue injury and are involved in cytoprotection. Arachidonylethanolamide (AEA) is a well characterized NAE that is an endogenous ligand at cannabinoid and vanilloid receptors, but it exists in small quantities relative to other NAE types. The abundance of other NAE species, such as palmitoylethanolamine (PEA), together with their largely unknown function and receptors, has prompted us to examine the neuroprotective properties and mechanism of action of PEA. We hypothesized that PEA protects HT22 cells from oxidative stress and activates neuroprotective kinase signaling pathways. RESULTS: Indeed PEA protected HT22 cells from oxidative stress in part by mediating an increase in phosphorylated Akt (pAkt) and ERK1/2 immunoreactivity as well as pAkt nuclear translocation. These changes take place within a time frame consistent with neuroprotection. Furthermore, we determined that changes in pAkt immunoreactivity elicited by PEA were not mediated by activation of cannabinoid receptor type 2 (CB2), thus indicating a novel mechanism of action. These results establish a role for PEA as a neuroprotectant against oxidative stress, which occurs in a variety of neurodegenerative diseases. CONCLUSIONS: The results from this study reveal that PEA protects HT22 cells from oxidative stress and alters the localization and expression levels of kinases known to be involved in neuroprotection by a novel mechanism. Overall, these results identify PEA as a neuroprotectant with potential as a possible therapeutic agent in neurodegenerative diseases involving oxidative stress. PMID- 20003318 TI - Improved accuracy in flow mapping of congenital heart disease using stationary phantom technique. AB - BACKGROUND: Flow mapping by cardiovascular magnetic resonance has become the gold standard for non-invasively defining cardiac output (CO), shunt flow and regurgitation. Previous reports have highlighted the presence of inherent errors in flow mapping that are improved with the use of a stationary phantom control. To our knowledge, these studies have only been performed in healthy volunteers. RESULTS: We analyzed the variation in flow measurements made with and without stationary phantom correction in 31 patients with congenital heart disease. Variation in stroke volume (SV) measurements was seen in all vessels across all patient groups. The variation was largest when analyzing the right ventricular outflow tract (RVOT), with a range of absolute differences in SV from 0.2 to 70 ml and in CO from 0.02 to 4.8 L/min. In patients with repaired Tetrology of Fallot (ToF), the average ratio of pulmonary to systemic blood flow (Qp:Qs) was 1.18 without and 1.02 with phantom correction. Without performing phantom correction, 23% of the repaired ToF patients were classified as having a residual shunt as compared to 0% when flow mapping was performed with phantom correction. Similarly, in patients with known atrial level shunting (ASD/PAPVR) 20% of patients had no shunt when flow mapping was performed without phantom correction as compared to 0% with phantom correction. In patients with bicuspid aortic valves (BAV), the differences in the regurgitant fraction between measuring flow with and without phantom correction ranged from 0 to 30%, while the regurgitant fraction in the RVOT of ToF patients varied by as much as 31%. CONCLUSION: The impact of inherent errors in CMR flow mapping should not be underestimated. While the variation across a population may not display a significant trend, for any individual patient it can be quite large. Failure to correct for such variation can lead to clinically significant misinterpretation of flow data. The use of the stationary phantom correction technique appears to improve accuracy both in normal patients as well as those with congenital heart disease. PMID- 20003319 TI - Identification of known and novel pancreas genes expressed downstream of Nkx2.2 during development. AB - BACKGROUND: The homeodomain containing transcription factor Nkx2.2 is essential for the differentiation of pancreatic endocrine cells. Deletion of Nkx2.2 in mice leads to misspecification of islet cell types; insulin-expressing beta cells and glucagon-expressing alpha cells are replaced by ghrelin-expressing cells. Additional studies have suggested that Nkx2.2 functions both as a transcriptional repressor and activator to regulate islet cell formation and function. To identify genes that are potentially regulated by Nkx2.2 during the major wave of endocrine and exocrine cell differentiation, we assessed gene expression changes that occur in the absence of Nkx2.2 at the onset of the secondary transition in the developing pancreas. RESULTS: Microarray analysis identified 80 genes that were differentially expressed in e12.5 and/or e13.5 Nkx2.2-/- embryos. Some of these genes encode transcription factors that have been previously identified in the pancreas, clarifying the position of Nkx2.2 within the islet transcriptional regulatory pathway. We also identified signaling factors and transmembrane proteins that function downstream of Nkx2.2, including several that have not previously been described in the pancreas. Interestingly, a number of known exocrine genes are also misexpressed in the Nkx2.2-/- pancreas. CONCLUSIONS: Expression profiling of Nkx2.2-/- mice during embryogenesis has allowed us to identify known and novel pancreatic genes that function downstream of Nkx2.2 to regulate pancreas development. Several of the newly identified signaling factors and transmembrane proteins may function to influence islet cell fate decisions. These studies have also revealed a novel function for Nkx2.2 in maintaining appropriate exocrine gene expression. Most importantly, Nkx2.2 appears to function within a complex regulatory loop with Ngn3 at a key endocrine differentiation step. PMID- 20003320 TI - Destructive arthritis in a patient with chikungunya virus infection with persistent specific IgM antibodies. AB - BACKGROUND: Chikungunya fever is an emerging arboviral disease characterized by an algo-eruptive syndrome, inflammatory polyarthralgias, or tenosynovitis that can last for months to years. Up to now, the pathophysiology of the chronic stage is poorly understood. CASE PRESENTATION: We report the first case of CHIKV infection with chronic associated rheumatism in a patient who developed progressive erosive arthritis with expression of inflammatory mediators and persistence of specific IgM antibodies over 24 months following infection. CONCLUSIONS: Understanding the specific features of chikungunya virus as well as how the virus interacts with its host are essential for the prevention, treatment or cure of chikungunya disease. PMID- 20003321 TI - Whole pelvic helical tomotherapy for locally advanced cervical cancer: technical implementation of IMRT with helical tomotherapy. AB - BACKGROUND: To review the experience and to evaluate the treatment plan of using helical tomotherapy (HT) for the treatment of cervical cancer. METHODS: Between November 1st, 2006 and May 31, 2009, 10 cervical cancer patients histologically confirmed were enrolled. All of the patients received definitive concurrent chemoradiation (CCRT) with whole pelvic HT (WPHT) followed by brachytherapy. During WPHT, all patients were treated with cisplatin, 40 mg/m2 intravenously weekly. Toxicity of treatment was scored according to the Common Terminology Criteria for Adverse Events v3.0 (CTCAE v3.0). RESULTS: The mean survival was 25 months (range, 3 to 27 months). The actuarial overall survival, disease-free survival, locoregional control and distant metastasis-free rates at 2 years were 67%, 77%, 90% and 88%, respectively. The average of uniformity index and conformal index was 1.06 and 1.19, respectively. One grade 3 of acute toxicity for diarrhea, thrombocytopenia and three grade 3 leucopenia were noted during CCRT. Only one grade 3 of subacute toxicity for thrombocytopenia was noted. There were no grade 3 or 4 subacute toxicities of anemia, leucopenia, genitourinary or gastrointestinal effects. Compared with conventional whole pelvic radiation therapy (WPRT), WPHT decreases the mean dose to rectum, bladder and intestines successfully. CONCLUSION: HT provides feasible clinical outcomes in locally advanced cervical cancer patients. Long-term follow-up and enroll more locally advanced cervical carcinoma patients by limiting bone marrow radiation dose with WPHT technique is warranted. PMID- 20003322 TI - A replication study of the association between the IL12B promoter allele CTCTAA and susceptibility to cerebral malaria in Thai population. AB - BACKGROUND: Interleukin-12 (IL-12), a heterodimeric cytokine composed of p35 and p40 subunits, has been thought to play an important role in the pathogenesis of malaria. The IL-12p40 subunit is encoded by the IL12B gene. An IL12B promoter allele, CTCTAA, at rs17860508 has been reported to be associated with susceptibility to cerebral malaria in African populations. However, this association has not so far been replicated in non-African populations. METHODS: To examine whether the CTCTAA allele is associated with susceptibility to cerebral malaria in Asian populations, 303 Thai patients with Plasmodium falciparum malaria (109 cerebral malaria and 194 mild malaria patients) were genotyped for rs17860508 by PCR-direct sequencing. RESULTS: The CTCTAA allele showed a significant association with susceptibility to cerebral malaria in the Thai population (allelic OR = 1.37; one sided P-value = 0.030). CONCLUSIONS: The existence of a significant association between the CTCTAA allele and susceptibility to cerebral malaria was confirmed in Southeast Asian population, which was previously reported in African populations. PMID- 20003323 TI - RANKL inhibition by osteoprotegerin prevents bone loss without affecting local or systemic inflammation parameters in two rat arthritis models: comparison with anti-TNFalpha or anti-IL-1 therapies. AB - INTRODUCTION: Rat adjuvant-induced arthritis (AIA) and collagen-induced arthritis (CIA) feature bone loss and systemic increases in TNFalpha, IL-1beta, and receptor activator of NF-kappaB ligand (RANKL). Anti-IL-1 or anti-TNFalpha therapies consistently reduce inflammation in these models, but systemic bone loss often persists. RANKL inhibition consistently prevents bone loss in both models without reducing joint inflammation. Effects of these therapies on systemic markers of bone turnover and inflammation have not been directly compared. METHODS: Lewis rats with established AIA or CIA were treated for 10 days (from day 4 post onset) with either PBS (Veh), TNFalpha inhibitor (pegsunercept), IL-1 inhibitor (anakinra), or RANKL inhibitor (osteoprotegerin (OPG)-Fc). Local inflammation was evaluated by monitoring hind paw swelling. Bone mineral density (BMD) of paws and lumbar vertebrae was assessed by dual X-ray absorptiometry. Markers and mediators of bone resorption (RANKL, tartrate resistant acid phosphatase 5b (TRACP 5B)) and inflammation (prostaglandin E2 (PGE2), acute-phase protein alpha-1-acid glycoprotein (alpha1AGP), multiple cytokines) were measured in serum (day 14 post onset). RESULTS: Arthritis progression significantly increased paw swelling and ankle and vertebral BMD loss. Anti-TNFalpha reduced paw swelling in both models, and reduced ankle BMD loss in AIA rats. Anti-IL-1 decreased paw swelling in CIA rats, and reduced ankle BMD loss in both models. Anti-TNFalpha and anti-IL-1 failed to prevent vertebral BMD loss in either model. OPG-Fc reduced BMD loss in ankles and vertebrae in both models, but had no effect on paw swelling. Serum RANKL was elevated in AIA-Veh and CIA-Veh rats. While antiTNFalpha and anti-IL-1 partially normalized serum RANKL without any changes in serum TRACP 5B, OPG-Fc treatment reduced serum TRACP 5B by over 90% in both CIA and AIA rats. CIA-Veh and AIA-Veh rats had increased serum alpha1AGP, IL-1beta, IL-8 and chemokine (C-C motif) ligand 2 (CCL2), and AIA-Veh rats also had significantly greater serum PGE2, TNFalpha and IL-17. Anti TNFalpha reduced systemic alpha1AGP, CCL2 and PGE2 in AIA rats, while anti-IL-1 decreased systemic alpha1AGP, IL-8 and PGE2. In contrast, RANKL inhibition by OPG Fc did not lessen systemic cytokine levels in either model. CONCLUSIONS: Anti TNFalpha or anti-IL-1 therapy inhibited parameters of local and systemic inflammation, and partially reduced local but not systemic bone loss in AIA and CIA rats. RANKL inhibition prevented local and systemic bone loss without significantly inhibiting local or systemic inflammatory parameters. PMID- 20003324 TI - Evidence for an association of HLA-DRB1*15 and DRB1*09 with leprosy and the impact of DRB1*09 on disease onset in a Chinese Han population. AB - BACKGROUND: Human leukocyte antigens (HLAs) have been proposed to modulate the immune response to Mycobacterium leprae. The association of HLA-DRB1 with leprosy has been reported in several populations, but not in a Chinese population. METHODS: The polymerase chain reaction-sequence-specific oligonucleotide probe with Luminex100 (PCR-SSOP-Luminex) method was used to genotype HLA-DRB1 alleles in 305 leprosy patients and 527 healthy control individuals. RESULTS: The HLA DRB1*15 allele was significantly more prevalent among leprosy patients than healthy controls, whereas the frequency of the HLA-DRB1*09 allele was lower among leprosy patients, especially those with early-onset disease. CONCLUSION: HLA-DRB1 alleles are associated with leprosy susceptibility in a Chinese population. The HLA-DRB1*09 allele was found to be protective exclusively in a subset of early onset leprosy patients. PMID- 20003325 TI - Inhibition of SOC/Ca2+/NFAT pathway is involved in the anti-proliferative effect of sildenafil on pulmonary artery smooth muscle cells. AB - BACKGROUND: Sildenafil, a potent phosphodiesterase type 5 (PDE5) inhibitor, has been proposed as a treatment for pulmonary arterial hypertension (PAH). The mechanism of its anti-proliferative effect on pulmonary artery smooth muscle cells (PASMC) is unclear. Nuclear translocation of nuclear factor of activated T cells (NFAT) is thought to be involved in PASMC proliferation and PAH. Increase in cytosolic free [Ca2+] ([Ca2+]i) is a prerequisite for NFAT nuclear translocation. Elevated [Ca2+]i in PASMC of PAH patients has been demonstrated through up-regulation of store-operated Ca2+ channels (SOC) which is encoded by the transient receptor potential (TRP) channel protein. Thus we investigated if: 1) up-regulation of TRPC1 channel expression which induces enhancement of SOC mediated Ca2+ influx and increase in [Ca2+]i is involved in hypoxia-induced PASMC proliferation; 2) hypoxia-induced promotion of [Ca2+]i leads to nuclear translocation of NFAT and regulates PASMC proliferation and TRPC1 expression; 3) the anti-proliferative effect of sildenafil is mediated by inhibition of this SOC/Ca2+/NFAT pathway. METHODS: Human PASMC were cultured under hypoxia (3% O2) with or without sildenafil treatment for 72 h. Cell number and cell viability were determined with a hemocytometer and MTT assay respectively. [Ca2+]i was measured with a dynamic digital Ca2+ imaging system by loading PASMC with fura 2 AM. TRPC1 mRNA and protein level were detected by RT-PCR and Western blotting respectively. Nuclear translocation of NFAT was determined by immunofluoresence microscopy. RESULTS: Hypoxia induced PASMC proliferation with increases in basal [Ca2+]i and Ca2+ entry via SOC (SOCE). These were accompanied by up-regulation of TRPC1 gene and protein expression in PASMC. NFAT nuclear translocation was significantly enhanced by hypoxia, which was dependent on SOCE and sensitive to SOC inhibitor SKF96365 (SKF), as well as cGMP analogue, 8-brom-cGMP. Hypoxia induced PASMC proliferation and TRPC1 up-regulation were inhibited by SKF and NFAT blocker (VIVIT and Cyclosporin A). Sildenafil treatment ameliorated hypoxia induced PASMC proliferation and attenuated hypoxia-induced enhancement of basal [Ca2+]i, SOCE, up-regulation of TRPC1 expression, and NFAT nuclear translocation. CONCLUSION: The SOC/Ca2+/NFAT pathway is, at least in part, a downstream mediator for the anti-proliferative effect of sildenafil, and may have therapeutic potential for PAH treatment. PMID- 20003326 TI - Promoter methylation of IGFBP-3 and p53 expression in ovarian endometrioid carcinoma. AB - BACKGROUND: Insulin-like growth factor binding protein (IGFBP-3) is an antiproliferative, pro-apoptotic and invasion suppressor protein which is transcriptionally regulated by p53. Promoter methylation has been linked to gene silencing and cancer progression. We studied the correlation between IGFBP-3 and p53 expression as well as IGFBP-3 promoter methylation in ovarian endometrioid carcinoma (OEC) by immunohistochemical staining and quantitative methylation specific PCR (qMSP). Additionally, we assessed the molecular regulatory mechanism of wild type (wt) p53 on IGFBP-3 expression using two subclones of OEC, the OVTW59-P0 (low invasive) and P4 (high invasive) sublines. RESULTS: In 60 cases of OEC, 40.0% showed lower IGFBP-3 expression which was significantly correlated with higher IGFBP-3 promoter methylation. p53 overexpression was detected in 35.0% of OEC and was unrelated to clinical outcomes and IGFBP-3. By Kaplan-Meier analysis, patients with lower IGFBP-3, higher IGFBP-3 promoter methylation, and normal p53 were associated most significantly with lower survival rates. In OEC cell line, IGFBP-3 expression was correlated with IGFBP-3 promoter methylation. IGFBP-3 expression was restored after treatment with a DNA methy-transferase inhibitors (5-aza-deoxycytidine) and suppressed by a p53 inhibitor (pifithrin alpha). The putative p53 regulatory sites on the promoter of IGFBP-3 were identified at -210, -206, -183 and -179 bases upstream of the transcription start site. Directed mutagenesis at these sites quantitatively reduced the transcription activity of IGFBP-3. CONCLUSION: Our data suggests that IGFBP-3 silencing through IGFBP-3 promoter methylation in the absence of p53 overexpression is associated with cancer progression. These results support a potential role of IGFBP-3 methylation in the carcinogenesis of OEC. PMID- 20003327 TI - Fall-related injuries in a nursing home setting: is polypharmacy a risk factor? AB - BACKGROUND: Polypharmacy is regarded as an important risk factor for fallingand several studies and meta-analyses have shown an increased fall risk in users of diuretics, type 1a antiarrhythmics, digoxin and psychotropic agents. In particular, recent evidence has shown that fall risk is associated with the use of polypharmacy regimens that include at least one established fall risk increasing drug, rather than with polypharmacy per se. We studied the role of polypharmacy and the role of well-known fall risk-increasing drugs on the incidence of injurious falls. METHODS: A retrospective observational study was carried out in a population of elderly nursing home residents. An unmatched, post stratification design for age class, gender and length of stay was adopted. In all, 695 falls were recorded in 293 residents. RESULTS: 221 residents (75.4%) were female and 72 (24.6%) male, and 133 (45.4%) were recurrent fallers. 152 residents sustained no injuries when they fell, whereas injuries were sustained by 141: minor in 95 (67.4%) and major in 46 (32.6%). Only fall dynamics (p = 0.013) and drugs interaction between antiarrhythmic or antiparkinson class and polypharmacy regimen (> or =7 medications) seem to represent a risk association for injuries (p = 0.024; OR = 4.4; CI 95% 1.21 - 15.36). CONCLUSION: This work reinforces the importance of routine medication reviews, especially in residents exposed to polypharmacy regimens that include antiarrhythmics or antiparkinson drugs, in order to reduce the risk of fall-related injuries during nursing home stays. PMID- 20003328 TI - A rapid malaria appraisal in the Venezuelan Amazon. AB - BACKGROUND: While the federal state of Amazonas bears the highest risk for malaria in Venezuela (2007: 68.4 cases/1000 inhabitants), little comprehensive information about the malaria situation is available from this area. The purpose of this rapid malaria appraisal (RMA) was to provide baseline data about malaria and malaria control in Amazonas. METHODS: The RMA methodology corresponds to a rapid health impact assessment (HIA) as described in the 1999 Gothenburg consensus. In conjunction with the actors of the malaria surveillance system, all useful data and information, which were accessible within a limited time-frame of five visits to Amazonas, were collected, analysed and interpreted. RESULTS: Mortality from malaria is low (< 1 in 105) and slide positivity rates have stayed at the same level for the last two decades (15% +/- 6% (SD)). Active case detection accounts for ca. 40% of slides taken. The coverage of the censured population with malaria notification points (NPs) has been achieved in recent years. The main parasite is Plasmodium vivax (84% of cases). The proportion of Plasmodium falciparum is on the decline, possibly driven by the introduction of cost-free artemisinin-based combination therapy (ACT) (1988: 33.4%; 2007: 15.4%). Monitoring and documentation is complete, systematic and consistent, but poorly digitalized. Malaria transmission displayed a visible lag behind rainfall in the capital municipality of Atures, but not in the other municipalities. In comparison to reference microscopy, quality of field microscopy and rapid diagnostic tests (RDTs) is suboptimal (kappa < 0.75). Hot spots of malaria risk were seen in some indigenous ethnic groups. Conflicting strategies in respect of training of community health workers (CHW) and the introduction of new diagnostic tools (RDTs) were observed. CONCLUSION: Malaria control is possible, even in tropical rain forest areas, if the health system is working adequately. Interventions have to be carefully designed and the features of the particular local Latin American context considered. PMID- 20003329 TI - Role of p53 mutation in the effect of boron neutron capture therapy on oral squamous cell carcinoma. AB - BACKGROUND: Boron neutron capture therapy (BNCT) is a selective radiotherapy, being effective for the treatment of even advanced malignancies in head and neck regions as well as brain tumors and skin melanomas. To clarify the role of p53 gene, the effect of BNCT on oral squamous cell carcinoma (SCC) cells showing either wild- (SAS/neo) or mutant-type (SAS/mp53) p53 was examined. METHODS: Cells were exposed to neutron beams in the presence of boronophenylalanine (BPA) at Kyoto University Research Reactor. Treated cells were monitored for modulations in colony formation, proliferation, cell cycle, and expression of cell cycle associated proteins. RESULTS: When SAS/neo and SAS/mp53 cells were subjected to BNCT, more suppressive effects on colony formation and cell viability were observed in SAS/neo compared with SAS/mp53 cells. Cell cycle arrest at the G1 checkpoint was observed in SAS/neo, but not in SAS/mp53. Apoptotic cells increased from 6 h after BNCT in SAS/neo and 48 h in SAS/mp53 cells. The expression of p21 was induced in SAS/neo only, but G2 arrest-associated proteins including Wee1, cdc2, and cyclin B1 were altered in both cell lines. CONCLUSION: These results indicate that oral SCC cells with mutant-type are more resistant to BNCT than those with wild-type p53, and that the lack of G1 arrest and related apoptosis may contribute to the resistance. At a physical dose affecting the cell cycle, BNCT inhibits oral SCC cells in p53-dependent and -independent manners. PMID- 20003330 TI - Intraventricular infusion of hyperosmolar dextran induces hydrocephalus: a novel animal model of hydrocephalus. AB - BACKGROUND: Popular circulation theory of hydrocephalus assumes that the brain is impermeable to cerebrospinal fluid (CSF), and is therefore incapable of absorbing the CSF accumulating within the ventricles. However, the brain parenchyma is permeable to water due to the presence of specific ion channels as well as aquaporin channels. Thus, the movement of water into and out of the ventricles may be determined by the osmotic load of the CSF. If osmotic load determines the aqueous content of CSF in this manner, it is reasonable to hypothesize that hydrocephalus may be precipitated by pathologies and/or insults that produce sustained elevations of osmotic content within the ventricles. METHODS: We investigated this hypothesis by manipulating the osmotic content of CSF and assaying the development of hydrocephalus in the rat brain. This was achieved by continuously infusing artificial CSF (negative control; group I), fibroblast growth factor (FGF2) solution (positive control; group II) and hyperosmotic dextran solutions (10 KD and 40 KD as experimental solutions: groups III and IV) for 12 days at 0.5 muL/h. The osmolality of the fluid infused was 307, 664, 337 and 328 mOsm/L in Groups I, II, III and IV, respectively. Magnetic resonance imaging (MRI) was used to evaluate the ventricular volumes. Analysis of variance (ANOVA) with pairwise group comparisons was done to assess the differences in ventricular volumes among the four groups. RESULTS: Group I had no hydrocephalus. Group II, group III and group IV animals exhibited significant enlargement of the ventricles (hydrocephalus) compared to group I. There was no statistically significant difference in the size of the ventricles between groups II, III and IV. None of the animals with hydrocephalus had obstruction of the aqueduct or other parts of CSF pathways on MRI. CONCLUSION: Infusing hyperosmolar solutions of dextran, or FGF into the ventricles chronically, resulted in ventricular enlargement. These solutions increase the osmotic load in the ventricles. Water influx (through the choroid plexus CSF secretion and/or through the brain) into the ventricles to normalize this osmotic gradient results in hydrocephalus. We need to revise the popular theory of how fluid accumulates in the ventricles at least in some forms of hydrocephalus. PMID- 20003331 TI - The tracking of dietary intakes of children and adolescents in Sweden over six years: the European Youth Heart Study. AB - BACKGROUND: The stability of dietary habits through various life-stages is not well understood. A better understanding of the tracking of diet over time could have implications for health promotion as well as for the planning of nutritional epidemiology studies. We examined the stability of dietary intakes of children and adolescents over six years. METHODS: As part of the European Youth Heart Study, in 1998-9, a 24-h dietary recall was performed on over one thousand 9- and 15-year-olds in Sweden. In 2004-5, 40% returned to the follow-up study. These 452 subjects (273 15- and 179 21-year-olds) were assigned to age- and gender-specific tertiles of intakes of food groups, energy, selected nutrients and energy density (low, mid and high) at each time point. The agreement between the classification of subjects into tertiles at both time points was examined using Cohen's weighted kappa and other stability coefficients. We included a dropout analysis and considered the effect that energy mis-reporting might have on our results. RESULTS: Fair tracking was seen between childhood and adolescence for the milk, fil and yoghurt food group (kappa = 0.30), and between adolescence and young adulthood for fruit (kappa = 0.24). Slight tracking was observed for most other food groups and fair to slight tracking for all nutrients studied. Only membership of the high milk, fil and yoghurt tertile could be predicted from membership at baseline, in children. Excluding potential energy mis-reporters did not affect the results. CONCLUSIONS: Despite the long time between data collections, and the method of dietary data collection used, evidence for slight tracking was observed for most food groups and nutrients over these six years. PMID- 20003332 TI - Methylprednisolone inhibits IFN-gamma and IL-17 expression and production by cells infiltrating central nervous system in experimental autoimmune encephalomyelitis. AB - BACKGROUND: Glucocorticoids have been shown to be effective in the treatment of autoimmune diseases of the CNS such as multiple sclerosis and its animal model, experimental autoimmune encephalomyelitis (EAE). However, the mechanisms and the site of glucocorticoids' actions are still not completely defined. The aim of this study was to investigate the in vivo effect of the synthetic glucocorticoid methylprednisolone (MP) on the expression and production of proinflammatory cytokines interferon (IFN)-gamma and interleukin (IL)-17 by cells infiltrating CNS tissue. METHODS: Experimental autoimmune encephalomyelitis was induced in Dark Agouti (DA) rats by immunization with rat spinal cord homogenate mixed with adjuvants. Commencing on the day when the first EAE signs appeared, DA rats were injected daily for 3 days with MP and/or RU486, an antagonist of glucocorticoid receptor. Cytokine production and gene expression in CNS-infiltrating cells and lymph node cells were measured using ELISA and real time PCR, respectively. RESULTS: Treatment of rats with MP ameliorated EAE, and the animals recovered without relapses. Further, MP inhibited IFN-gamma and IL-17 expression and production in cells isolated from the CNS of DA rats with EAE after the last injection of MP. The observed effect of MP in vivo treatment was not mediated through depletion of CD4+ T cells among CNS infiltrating cells, or through induction of their apoptosis within the CNS. Finally, the glucocorticoid receptor antagonist RU486 prevented the inhibitory effect of MP on IFN-gamma and IL-17 production both in vitro and in vivo, thus indicating that the observed effects of MP were mediated through glucocorticoid receptor-dependent mechanisms. CONCLUSION: Taken together, these results demonstrate that amelioration of EAE by exogenous glucocorticoids might be, at least partly, ascribed to the limitation of effector cell functions in the target tissue. PMID- 20003333 TI - Laparoscopic retrograde (fundus first) cholecystectomy. AB - BACKGROUND: Retrograde ("fundus first") dissection is frequently used in open cholecystectomy and although feasible in laparoscopic cholecystectomy (LC) it has not been widely practiced. LC is most simply carried out using antegrade dissection with a grasper to provide cephalad fundic traction. A series is presented to investigate the place of retrograde dissection in the hands of an experienced laparoscopic surgeon using modern instrumentation. METHODS: A prospective record of all LCs carried out by an experienced laparoscopic surgeon following his appointment in Bristol in 2004 was examined. Retrograde dissection was resorted to when difficulties were encountered with exposure and/or dissection of Calot's triangle. RESULTS: 1041 LCs were carried out including 148 (14%) emergency operations and 131 (13%) associated bile duct explorations. There were no bile duct injuries although conversion to open operation was required in six patients (0.6%). Retrograde LC was attempted successfully in 11 patients (1.1%). The age ranged from 28 to 80 years (mean 61) and there were 7 males. Indications were; fibrous, contracted gallbladder 7, Mirizzi syndrome 2 and severe kyphosis 2. Operative photographs are included to show the type of case where it was needed and the technique used. Postoperative stay was 1/2 to 5 days (mean 2.2) with no delayed sequelae on followup. Histopathology showed; chronic cholecystitis 7, xanthogranulomatous cholecystitis 3 and acute necrotising cholecystitis 1. CONCLUSIONS: In this series, retrograde laparoscopic dissection was necessary in 1.1% of LCs and a liver retractor was needed in 9 of the 11 cases. This technique does have a place and should be in the armamentarium of the laparoscopic surgeon. PMID- 20003334 TI - A rare case of acute lymphoblastic leukaemia with hemophilia A. AB - A rare case of Acute lymphoblastic leukemia with hemophillia in a 12 year old boy is presented in the article. Patient was known case of hemophillia (factor VIII deficiency). He was diagnosed as a case of ALL based on bone marrow examination and immunophenotypic study. Patient was treated as per Children Cancer group guidelines. The main aim of reporting this rare association lies in developing treatment strategies in preventing life threatening bleeding due to this rare association which though may be accidental but need further research. PMID- 20003335 TI - APRIL is a novel clinical chemo-resistance biomarker in colorectal adenocarcinoma identified by gene expression profiling. AB - BACKGROUND: 5-Fluorouracil(5FU) and oral analogues, such as capecitabine, remain one of the most useful agents for the treatment of colorectal adenocarcinoma. Low toxicity and convenience of administration facilitate use, however clinical resistance is a major limitation. Investigation has failed to fully explain the molecular mechanisms of resistance and no clinically useful predictive biomarkers for 5FU resistance have been identified. We investigated the molecular mechanisms of clinical 5FU resistance in colorectal adenocarcinoma patients in a prospective biomarker discovery project utilising gene expression profiling. The aim was to identify novel 5FU resistance mechanisms and qualify these as candidate biomarkers and therapeutic targets. METHODS: Putative treatment specific gene expression changes were identified in a transcriptomics study of rectal adenocarcinomas, biopsied and profiled before and after pre-operative short course radiotherapy or 5FU based chemo-radiotherapy, using microarrays. Tumour from untreated controls at diagnosis and resection identified treatment independent gene expression changes. Candidate 5FU chemo-resistant genes were identified by comparison of gene expression data sets from these clinical specimens with gene expression signatures from our previous studies of colorectal cancer cell lines, where parental and daughter lines resistant to 5FU were compared. A colorectal adenocarcinoma tissue microarray (n = 234, resected tumours) was used as an independent set to qualify candidates thus identified. RESULTS: APRIL/TNFSF13 mRNA was significantly upregulated following 5FU based concurrent chemo-radiotherapy and in 5FU resistant colorectal adenocarcinoma cell lines but not in radiotherapy alone treated colorectal adenocarcinomas. Consistent with APRIL's known function as an autocrine or paracrine secreted molecule, stromal but not tumour cell protein expression by immunohistochemistry was correlated with poor prognosis (p = 0.019) in the independent set. Stratified analysis revealed that protein expression of APRIL in the tumour stroma is associated with survival in adjuvant 5FU treated patients only (n = 103, p < 0.001), and is independently predictive of lack of clinical benefit from adjuvant 5FU [HR 6.25 (95%CI 1.48-26.32), p = 0.013]. CONCLUSIONS: A combined investigative model, analysing the transcriptional response in clinical tumour specimens and cancers cell lines, has identified APRIL, a novel chemo-resistance biomarker with independent predictive impact in 5FU-treated CRC patients, that may represent a target for novel therapeutics. PMID- 20003336 TI - Income level and chronic ambulatory care sensitive conditions in adults: a multicity population-based study in Italy. AB - BACKGROUND: A relationship between quality of primary health care and preventable hospitalizations has been described in the US, especially among the elderly. In Europe, there has been a recent increase in the evaluation of Ambulatory Care Sensitive Conditions (ACSC) as an indicator of health care quality, but evidence is still limited. The aim of this study was to determine whether income level is associated with higher hospitalization rates for ACSC in adults in a country with universal health care coverage. METHODS: From the hospital registries in four Italian cities (Turin, Milan, Bologna, Rome), we identified 9384 hospital admissions for six chronic conditions (diabetes, hypertension, congestive heart failure, angina pectoris, chronic obstructive pulmonary disease, and asthma) among 20-64 year-olds in 2000. Case definition was based on the ICD-9-CM coding algorithm suggested by the Agency for Health Research and Quality - Prevention Quality Indicators. An area-based (census block) income index was used for each individual. All hospitalization rates were directly standardised for gender and age using the Italian population. Poisson regression analysis was performed to assess the relationship between income level (quintiles) and hospitalization rates (RR, 95% CI) separately for the selected conditions controlling for age, gender and city of residence. RESULTS: Overall, the ACSC age-standardized rate was 26.1 per 10.000 inhabitants. All conditions showed a statistically significant socioeconomic gradient, with low income people being more likely to be hospitalized than their well off counterparts. The association was particularly strong for chronic obstructive pulmonary disease (level V low income vs. level I high income RR = 4.23 95%CI 3.37-5.31) and for congestive heart failure (RR = 3.78, 95% CI = 3.09-4.62). With the exception of asthma, males were more vulnerable to ACSC hospitalizations than females. The risks were higher among 45-64 year olds than in younger people. CONCLUSIONS: The socioeconomic gradient in ACSC hospitalization rates confirms the gap in health status between social groups in our country. Insufficient or ineffective primary care is suggested as a plausible additional factor aggravating inequality. This finding highlights the need for improving outpatient care programmes to reduce the excess of unnecessary hospitalizations among poor people. PMID- 20003337 TI - Activation of Retinoid X Receptor increases dopamine cell survival in models for Parkinson's disease. AB - BACKGROUND: Parkinson's disease (PD) is caused by degeneration of dopamine (DA) neurons in the ventral midbrain (vMB) and results in severely disturbed regulation of movement. The disease inflicts considerable suffering for the affected and their families. Today, the opportunities for pharmacological treatment are meager and new technologies are needed. Previous studies have indicated that activation of the nuclear receptor Retinoid X Receptor (RXR) provides trophic support for DA neurons. Detailed investigations of these neurotrophic effects have been hampered by the lack of readily available DA neurons in vitro. The aim of this study was to further describe the potential neurotrophic actions of RXR ligands and, for this and future purposes, develop a suitable in vitro-platform using mouse embryonic stem cells (mESCs). RESULTS: We studied the potential neurotrophic effects of the RXR ligand LG100268 (LG268) and the RXR-Nurr1 ligand XCT0139508 (XCT) in neuronal cultures derived from rat primary vMB and mESCs. RXR ligands protect DA neurons from stress, such as that induced by the PD-modeling toxin 6-hydroxy dopamine (6-OHDA) and hypoxia, but not from stress induced by oxidative hydrogen peroxide (H2O2) or the excitotoxic agent kainic acid (KA). The neurotrophic effect is selective for DA neurons. DA neurons from rat primary vMB and mESCs behaved similarly, but the mESC-derived cultures contained a much higher fraction of DA cells and thus provided more accessible experimental conditions. CONCLUSIONS: RXR ligands rescue DA neurons from degeneration caused by the PD simulating 6-OHDA as well as hypoxia. Thus, RXR is a novel promising target for PD research. mESC-derived DA cells provide a valid and accessible in vitro-platform for studying PD inducing toxins and potential trophic agents. PMID- 20003338 TI - The Prostate Care Questionnaire for Carers (PCQ-C): reliability, validity and acceptability. AB - BACKGROUND: Patient experience is commonly monitored in evaluating and improving health care, but the experience of carers (partners/relatives/friends) is rarely monitored even though the role of carers can often be substantial. For carers to fulfil their role it is necessary to address their needs. This paper describes an evaluation of the reliability, validity and acceptability of the PCQ-C, a newly developed instrument designed to measure the experiences of carers of men with prostate cancer. METHODS: The reliability, acceptability and validity of the PCQ C were tested through a postal survey and interviews with carers. The PCQ-C was posted to 1087 prostate cancer patients and patients were asked to pass the questionnaire on to their carer. Non-responders received one reminder. To assess test-retest reliability, 210 carers who had responded to the questionnaire were resent it a second time three weeks later. A subsample of nine carers from patients attending one hospital took part in qualitative interviews to assess validity and acceptability of the PCQ-C. Acceptability to service providers was evaluated based on four hospitals' experiences of running a survey using the PCQ C. RESULTS: Questionnaires were returned by 514 carers (47.3%), and the majority of questions showed less than 10% missing data. Across the sections of the questionnaire internal consistency was high (Cronbach's alpha ranging from 0.80 to 0.89), and test-retest stability showed moderate to high stability (intraclass correlation coefficients ranging from 0.52 to 0.83). Interviews of carers indicated that the PCQ-C was valid and acceptable. Feedback from hospitals indicated that they found the questionnaire useful, and highlighted important considerations for its future use as part of quality improvement initiatives. CONCLUSIONS: The PCQ-C has been found to be acceptable to carers and service providers having been used successfully in hospitals in England. It is ready for use to measure the aspects of care that need to be addressed to improve the quality of prostate cancer care, and for research. PMID- 20003340 TI - Identification and characterization of intervening sequences within 23S rRNA genes from more than 200 Campylobacter isolates from seven species including atypical campylobacters. AB - BACKGROUND: Identification and characterization of intervening sequences (IVSs) within 23S rRNA genes from Campylobacter organisms including atypical campylobacters were carried out using two PCR primer pairs, designed to generate helix 25 and 45 regions. RESULTS: Only C. sputorum biovar sputorum LMG7975 and fecalis LMG8531, LMG8534 and LMG6728 of a total of 204 Campylobacter isolates (n = 56 C. jejuni; n = 11 C. coli; n = 33 C. fetus; n = 43 C. upsaliensis; n = 30 C. hyointestinalis; n = 4 C. sputorum biovar sputorum; n = 5 C. sputorum biovar fecalis; n = 5 C. sputorum biovar paraureolyticus; n = 10 C. concisus; n = 7 C. curvus) were shown to carry IVSs in helix 25 region. C. sputorum biovar fecalis LMG8531 and LMG8534, interestingly, carried two different kinds of the 23S rRNA genes with and without the IVS, respectively. Consequently, in a total of 265 isolates of 269, including 65 C. lari isolates examined previously, the absence of IVSs was identified in the helix 25 region. In the helix 45 region, all the C. hyointestinalis, C. sputorum and C. concisus isolates were shown not to carry any IVSs. However, the 30 of 56 C. jejuni isolates (54%), 5 of 11 C. coli (45%), 25 of 33 C. fetus (76%), 30 of 43 C. upsaliensis (70%) and 6 of 7 C. curvus (90%) were shown to carry IVSs. In C. jejuni and C. upsaliensis isolates, two different kinds of the 23S rRNA genes were also identified to occur with and without IVSs in the helix 45 region, respectively. CONCLUSIONS: Secondary structure models were also constructed with all the IVSs identified in the present study. In the purified RNA fractions from the isolates which carried the 16S or 23S rRNA genes with the IVSs, no 16S or 23S rRNA was evident, respectively. PMID- 20003339 TI - Analysis of cat oocyte activation methods for the generation of feline disease models by nuclear transfer. AB - BACKGROUND: Somatic cell nuclear transfer in cats offers a useful tool for the generation of valuable research models. However, low birth rates after nuclear transfer hamper exploitation of the full potential of the technology. Poor embryo development after activation of the reconstructed oocytes seems to be responsible, at least in part, for the low efficiency. The objective of this study was to characterize the response of cat oocytes to various stimuli in order to fine-tune existing and possibly develop new activation methods for the generation of cat disease models by somatic cell nuclear transfer. METHODS: First, changes in the intracellular free calcium concentration [Ca2+]i in the oocytes induced by a number of artificial stimuli were characterized. The stimuli included electroporation, ethanol, ionomycin, thimerosal, strontium-chloride and sodium (Na+)-free medium. The potential of the most promising treatments (with or without subsequent incubation in the presence of cycloheximide and cytochalasin B) to stimulate oocyte activation and support development of the resultant parthenogenetic embryos was then evaluated. Finally, the most effective methods were selected to activate oocytes reconstructed during nuclear transfer with fibroblasts from mucopolysaccharidosis I- and alpha-mannosidosis-affected cats. RESULTS: All treatments were able to elicit a [Ca2+]i elevation in the ooplasm with various characteristics. Pronuclear formation and development up to the blastocyst stage was most efficiently triggered by electroporation (60.5 +/- 2.9 and 11.5 +/- 1.7%) and the combined thimerosal/DTT treatment (67.7 +/- 1.8 and 10.6 +/- 1.9%); incubation of the stimulated oocytes with cycloheximide and cytochalasin B had a positive effect on embryo development. When these two methods were used to activate oocytes reconstructed during nuclear transfer, up to 84.9% of the reconstructed oocytes cleaved. When the 2 to 4-cell embryos (a total of 220) were transferred into 19 recipient females, 4 animals became pregnant. All of the fetuses developed from oocytes activated by electroporation followed by cycloheximide and cytochalasin B incubation; no fetal development was detected as a result of thimerosal/DTT activation. Although heartbeats were detected in two of the cloned fetuses, no term development occurred. CONCLUSION: Electroporation proved to be the most effective method for the activation of cat oocytes reconstructed by nuclear transfer. The combined thimerosal/DTT treatment followed by cycloheximide and cytochalasin B incubation triggered development effectively to the blastocyst stage; whether it is a viable option to stimulate term development of cloned cat embryos needs further investigations. PMID- 20003341 TI - Transcutaneous electrical nerve stimulation for the management of tennis elbow: a pragmatic randomized controlled trial: the TATE trial (ISRCTN 87141084). AB - BACKGROUND: Tennis elbow is a common and often extremely painful musculoskeletal condition, which has considerable impact on individuals as well as economic implications for healthcare utilization and absence from work. Many management strategies have been studied in clinical trials. Whilst corticosteroid injections offer short term pain relief, this treatment is unpleasant and is used with caution due to an associated high risk of pain recurrence in the long term. Systematic reviews conclude that there is no clear and effective treatment for symptoms of pain in the first 6 weeks of the condition. There is a clear need for an intervention that is acceptable to patients and provides them with effective short-term pain relief without increasing the risk of recurrence. Transcutaneous electrical nerve stimulation (TENS) is an inexpensive, non-invasive, non pharmacological form of analgesia that is commonly used in the treatment of pain. TENS has very few contraindications and is simple to apply. It also benefits from being patient controlled, thereby promoting self-management. This study aims to assess the effectiveness, in terms of pain relief, and cost-effectiveness of a self-management package of treatment that includes TENS. METHODS/DESIGN: The design of the study will be a two-group pragmatic randomized clinical trial. 240 participants aged 18 years and over with tennis elbow will be recruited from 20 30 GP practices in Staffordshire, UK. Participants are to be randomized on a 1:1 basis to receive either primary care management (standard GP consultation, medication, advice and education) or primary care management with the addition of TENS, over 6 weeks. Our primary outcome measure is average intensity of elbow pain in the past 24 hours (0-10 point numerical rating scale) at 6 weeks. Secondary outcomes include pain and limitation of function, global assessment of change, days of sick leave, illness perceptions, and overall health status. A cost-effectiveness analysis will also be performed. Patient adherence and satisfaction data will be collected at 6 weeks, 6 months and 12 months by postal questionnaire. A diary will also be completed for the first 2 weeks of treatment. Clinical effectiveness and cost-effectiveness analyses will be carried out using an intention-to-treat approach as the primary analysis. DISCUSSION: This paper presents detail on the rationale, design, methods and operational aspects of the trial. TRIAL REGISTRATION: Current Controlled Trials. ISRCTN87141084. PMID- 20003342 TI - Identification of a biomarker panel using a multiplex proximity ligation assay improves accuracy of pancreatic cancer diagnosis. AB - BACKGROUND: Pancreatic cancer continues to prove difficult to clinically diagnose. Multiple simultaneous measurements of plasma biomarkers can increase sensitivity and selectivity of diagnosis. Proximity ligation assay (PLA) is a highly sensitive technique for multiplex detection of biomarkers in plasma with little or no interfering background signal. METHODS: We examined the plasma levels of 21 biomarkers in a clinically defined cohort of 52 locally advanced (Stage II/III) pancreatic ductal adenocarcinoma cases and 43 age-matched controls using a multiplex proximity ligation assay. The optimal biomarker panel for diagnosis was computed using a combination of the PAM algorithm and logistic regression modeling. Biomarkers that were significantly prognostic for survival in combination were determined using univariate and multivariate Cox survival models. RESULTS: Three markers, CA19-9, OPN and CHI3L1, measured in multiplex were found to have superior sensitivity for pancreatic cancer vs. CA19-9 alone (93% vs. 80%). In addition, we identified two markers, CEA and CA125, that when measured simultaneously have prognostic significance for survival for this clinical stage of pancreatic cancer (p < 0.003). CONCLUSIONS: A multiplex panel assaying CA19-9, OPN and CHI3L1 in plasma improves accuracy of pancreatic cancer diagnosis. A panel assaying CEA and CA125 in plasma can predict survival for this clinical cohort of pancreatic cancer patients. PMID- 20003343 TI - The synthetic inhibitor of fibroblast growth factor receptor PD166866 controls negatively the growth of tumor cells in culture. AB - BACKGROUND: Many experimental data evidence that over-expression of various growth factors cause disorders in cell proliferation. The role of the Fibroblast Growth Factors (FGF) in growth control is indisputable: in particular, FGF1 and its tyrosine kinase receptor (FGFR1) act through a very complex network of mechanisms and pathways. In this work we have evaluated the antiproliferative activity effect of PD166866, a synthetic molecule inhibiting the tyrosin kinase action of FGFR1. METHODS: Cells were routinely grown in Dulbecco Modified Eagle's medium supplemented with newborn serum and a penicillin-streptomycin mixture.Cell viability was evaluated by Mosmann assay and by trypan blue staining. DNA damage was assessed by in situ fluorescent staining with Terminal Deoxynucleotidyl Transferase dUTP nick end labeling (TUNEL assay).Assessment of oxidative stress at membrane level was measured by quantitative analysis of the intra-cellular formation of malonyl-dialdheyde (MDA) deriving from the decomposition of poly unsaturated fatty acids.The expression of Poly-ADP-Ribose-Polymerase (PARP), consequent to DNA fragmentation, was evidenced by immuno-histochemistry utilizing an antibody directed against an N-terminal fragment of the enzyme. RESULTS: The bioactivity of the drug was investigated on Hela cells. Cytoxicity was assessed by the Mosmann assay and by vital staining with trypan blue. The target of the molecule is most likely the cell membrane as shown by the significant increase of the intracellular concentration of malonyl-dihaldheyde. The increase of this compound, as a consequence of the treatment with PD166866, is suggestive of membrane lipoperoxidation. The TUNEL assay gave a qualitative, though clear, indication of DNA damage. Furthermore we demonstrate intracellular accumulation of poly-ADP-ribose polymerase I. This enzyme is a sensor of nicks on the DNA strands and this supports the idea that treatment with the drug induces cell death. CONCLUSIONS: Data presented in this work show that PD166866 has clear antiproliferative effects. The negative control of cell proliferation may be exerted through the activation of the apoptotic pathway. The results of experiments addressing this specific point, such as: evaluation of DNA damage, lipoperoxidation of the cell membrane and increase of expression of PARP, an enzyme directly involved in DNA repair. Results suggest that cells exposed to PD16866 undergo apoptosis. However, concomitant modes of cell death cannot be ruled out. The possible use of this drug for therapeutic purposes is discussed. PMID- 20003344 TI - Filling gaps in PPAR-alpha signaling through comparative nutrigenomics analysis. AB - BACKGROUND: The application of high-throughput genomic tools in nutrition research is a widespread practice. However, it is becoming increasingly clear that the outcome of individual expression studies is insufficient for the comprehensive understanding of such a complex field. Currently, the availability of the large amounts of expression data in public repositories has opened up new challenges on microarray data analyses. We have focused on PPARalpha, a ligand activated transcription factor functioning as fatty acid sensor controlling the gene expression regulation of a large set of genes in various metabolic organs such as liver, small intestine or heart. The function of PPARalpha is strictly connected to the function of its target genes and, although many of these have already been identified, major elements of its physiological function remain to be uncovered. To further investigate the function of PPARalpha, we have applied a cross-species meta-analysis approach to integrate sixteen microarray datasets studying high fat diet and PPARalpha signal perturbations in different organisms. RESULTS: We identified 164 genes (MDEGs) that were differentially expressed in a constant way in response to a high fat diet or to perturbations in PPARs signalling. In particular, we found five genes in yeast which were highly conserved and homologous of PPARalpha targets in mammals, potential candidates to be used as models for the equivalent mammalian genes. Moreover, a screening of the MDEGs for all known transcription factor binding sites and the comparison with a human genome-wide screening of Peroxisome Proliferating Response Elements (PPRE), enabled us to identify, 20 new potential candidate genes that show, both binding site, both change in expression in the condition studied. Lastly, we found a non random localization of the differentially expressed genes in the genome. CONCLUSION: The results presented are potentially of great interest to resume the currently available expression data, exploiting the power of in silico analysis filtered by evolutionary conservation. The analysis enabled us to indicate potential gene candidates that could fill in the gaps with regards to the signalling of PPARalpha and, moreover, the non-random localization of the differentially expressed genes in the genome, suggest that epigenetic mechanisms are of importance in the regulation of the transcription operated by PPARalpha. PMID- 20003345 TI - Atrial fibrillation detection by heart rate variability in Poincare plot. AB - BACKGROUND: Atrial fibrillation (AFib) is one of the prominent causes of stroke, and its risk increases with age. We need to detect AFib correctly as early as possible to avoid medical disaster because it is likely to proceed into a more serious form in short time. If we can make a portable AFib monitoring system, it will be helpful to many old people because we cannot predict when a patient will have a spasm of AFib. METHODS: We analyzed heart beat variability from inter-beat intervals obtained by a wavelet-based detector. We made a Poincare plot using the inter-beat intervals. By analyzing the plot, we extracted three feature measures characterizing AFib and non-AFib: the number of clusters, mean stepping increment of inter-beat intervals, and dispersion of the points around a diagonal line in the plot. We divided distribution of the number of clusters into two and calculated mean value of the lower part by k-means clustering method. We classified data whose number of clusters is more than one and less than this mean value as non-AFib data. In the other case, we tried to discriminate AFib from non AFib using support vector machine with the other feature measures: the mean stepping increment and dispersion of the points in the Poincare plot. RESULTS: We found that Poincare plot from non-AFib data showed some pattern, while the plot from AFib data showed irregularly irregular shape. In case of non-AFib data, the definite pattern in the plot manifested itself with some limited number of clusters or closely packed one cluster. In case of AFib data, the number of clusters in the plot was one or too many. We evaluated the accuracy using leave one-out cross-validation. Mean sensitivity and mean specificity were 91.4% and 92.9% respectively. CONCLUSIONS: Because pulse beats of ventricles are less likely to be influenced by baseline wandering and noise, we used the inter-beat intervals to diagnose AFib. We visually displayed regularity of the inter-beat intervals by way of Poincare plot. We tried to design an automated algorithm which did not require any human intervention and any specific threshold, and could be installed in a portable AFib monitoring system. PMID- 20003346 TI - Gene socialization: gene order, GC content and gene silencing in Salmonella. AB - BACKGROUND: Genes of conserved order in bacterial genomes tend to evolve slower than genes whose order is not conserved. In addition, genes with a GC content lower than the GC content of the resident genome are known to be selectively silenced by the histone-like nucleoid structuring protein (H-NS) in Salmonella. RESULTS: In this study, we use a comparative genomics approach to demonstrate that in Salmonella, genes whose order is not conserved (or genes without homologs) in closely related bacteria possess a significantly lower average GC content in comparison to genes that preserve their relative position in the genome. Moreover, these genes are more frequently targeted by H-NS than genes that have conserved their genomic neighborhood. We also observed that duplicated genes that do not preserve their genomic neighborhood are, on average, under less selective pressure. CONCLUSIONS: We establish a strong association between gene order, GC content and gene silencing in a model bacterial species. This analysis suggests that genes that are not under strong selective pressure (evolve faster than others) in Salmonella tend to accumulate more AT-rich mutations and are eventually silenced by H-NS. Our findings may establish new approaches for a better understanding of bacterial genome evolution and function, using information from functional and comparative genomics. PMID- 20003347 TI - Visualization of anomalous origin and course of coronary arteries in 748 consecutive symptomatic patients by 64-slice computed tomography angiography. AB - BACKGROUND: Coronary artery anomalies (CAAs) are currently undergoing profound changes in understanding potentially pathophysiological mechanisms of disease. Aim of this study was to investigate the prevalence of anomalous origin and course of coronary arteries in consecutive symptomatic patients, who underwent cardiac 64-slice multidetector-row computed tomography angiography (MDCTA). METHODS: Imaging datasets of 748 consecutive symptomatic patients referred for cardiac MDCTA were analyzed and CAAs of origin and further vessel course were grouped according to a recently suggested classification scheme by Angelini et al. RESULTS: An overall of 17/748 patients (2.3%) showed CAA of origin and further vessel course. According to aforementioned classification scheme no Subgroup 1- (absent left main trunk) and Subgroup 2- (anomalous location of coronary ostium within aortic root or near proper aortic sinus of Valsalva) CAA were found. Subgroup 3 (anomalous location of coronary ostium outside normal "coronary" aortic sinuses) consisted of one patient with high anterior origin of both coronary arteries. The remaining 16 patients showed a coronary ostium at improper sinus (Subgroup 4). Latter group was subdivided into a right coronary artery arising from left anterior sinus with separate ostium (subgroup 4a; n = 7) and common ostium with left main coronary artery (subgroup 4b; n = 1). Subgroup 4c consisted of one patient with a single coronary artery arising from the right anterior sinus (RAS) without left circumflex coronary artery (LCX). In subgroup 4d, LCX arose from RAS (n = 7). CONCLUSIONS: Prevalence of CAA of origin and further vessel course in a symptomatic consecutive patient population was similar to large angiographic series, although these patients do not reflect general population. However, our study supports the use of 64-slice MDCTA for the identification and definition of CAA. PMID- 20003348 TI - Postembryonic development of transit amplifying neuroblast lineages in the Drosophila brain. AB - BACKGROUND: Specific dorsomedial (DM) neuroblast lineages of the Drosophila brain amplify their proliferation through generation of transit amplifying intermediate progenitor cells. Together, these DM neuroblast lineages comprise over 5,000 adult-specific neural cells and thus represent a substantial part of the brain. However, no information is currently available about the structure or function of any of the neural cells in these DM lineages. In this report we use MARCM-based clonal analysis together with immunocytochemical labeling techniques to investigate the type and fate of neural cells generated in the DM lineages. RESULTS: Genetic cell lineage-tracing and immunocytochemical marker analysis reveal that DM neuroblasts are multipotent progenitors that produce a set of postembryonic brain glia as well as a large number of adult-specific protocerebral neurons. During larval development the adult-specific neurons of each DM lineage form several spatially separated axonal fascicles, some of which project along larval brain commissural structures that are primordia of midline neuropile. By taking advantage of a specific Gal4 reporter line, the DM-derived neuronal cells can be identified and followed into early pupal stages. During pupal development the neurons of the DM lineages arborize in many parts of the brain and contribute to neuropile substructures of the developing central complex, such as the fan-shaped body, noduli and protocerebral bridge. CONCLUSIONS: Our findings provide cellular and molecular evidence for the fact that DM neuroblasts are multipotent progenitors; thus, they represent the first identified progenitor cells in the fly brain that have neuroglioblast functions during postembryonic development. Moreover, our results demonstrate that the adult-specific neurons of the DM lineages arborize widely in the brain and also make a major contribution to the developing central complex. These findings suggest that the amplification of proliferation that characterizes DM lineages may be an important requirement for generating the large number of neurons required in highly complex neuropile structures such as the central complex in the Drosophila brain. PMID- 20003349 TI - Improved tRNA prediction in the American house dust mite reveals widespread occurrence of extremely short minimal tRNAs in acariform mites. AB - BACKGROUND: Atypical tRNAs are functional minimal tRNAs, lacking either the D- or T-arm. They are significantly shorter than typical cloverleaf tRNAs. Widespread occurrence of atypical tRNAs was first demonstrated for secernentean nematodes and later in various arachnids. Evidence started to accumulate that tRNAs of certain acariform mites are even shorter than the minimal tRNAs of nematodes, raising the possibility that tRNAs lacking both D- and T-arms might exist in these organisms. The presence of cloverleaf tRNAs in acariform mites, particularly in the house dust mite genus Dermatophagoides, is still disputed. RESULTS: Mitochondrial tRNAs of Dermatophagoides farinae are minimal, atypical tRNAs lacking either the T- or D-arm. The size (49-62, 54.4 +/- 2.86 nt) is significantly (p = 0.019) smaller than in Caenorhabditis elegans (53-63, 56.3 +/- 2.30 nt), a model minimal tRNA taxon. The shortest tRNA (49 nt) in Dermatophagoides is approaching the length of the shortest known tRNAs (45-49 nt) described in other acariform mites. The D-arm is absent in these tRNAs, and the inferred T-stem is small (2-3 bp) and thermodynamically unstable, suggesting that it may not exist in reality. The discriminator nucleotide is probably not encoded and is added postranscriptionally in many Dermatophagoides tRNAs. CONCLUSIONS: Mitochondrial tRNAs of acariform mites are largely atypical, non-cloverleaf tRNAs. Among them, the shortest known tRNAs with no D-arm and a short and unstable T-arm can be inferred. While our study confirmed seven tRNAs in Dermatophagoides by limited EST data, further experimental evidence is needed to demonstrate extremely small and unusual tRNAs in acariform mites. PMID- 20003350 TI - Strategies for recruiting Hispanic women into a prospective cohort study of modifiable risk factors for gestational diabetes mellitus. AB - BACKGROUND: The purpose of this article was to describe effective strategies for recruitment of Hispanic women into a prospective cohort study of modifiable risk factors for gestational diabetes mellitus (GDM). Although Hispanic women have two to four times the risk of developing GDM compared with non-Hispanic white women, few GDM prevention studies have included Hispanic women. METHODS: The study was conducted in the ambulatory obstetrical practices of Baystate Medical Center located in a socioeconomically and ethnically diverse city in Massachusetts. The study employed a range of strategies to recruit Hispanic women based on a review of the literature as well as prior experience with the study population. RESULTS: Over a period of 32 months, a total of 851 Hispanic prenatal care patients were recruited. Among eligible women, 52.4% agreed to participate. Participants were young (70% <25 years), with low levels of education, and on public health insurance (81.5%); 88% were unmarried. Study design features such as use of bilingual recruiters, a flexible recruitment process, training recruiters to be culturally sensitive, use of culturally tailored materials, prescreening participants, participant compensation, seeking the cooperation of clinic staff, and continuous monitoring of recruitment goals emerged as important issues influencing recruitment. CONCLUSIONS: Findings suggest that investigators can successfully recruit pregnant women from ethnic minority groups of low socioeconomic status into observational studies. The study provides culturally appropriate recruitment strategies useful for practice-based settings recruiting Hispanic research participation. PMID- 20003351 TI - Co-bedding as a Comfort measure For Twins undergoing painful procedures (CComForT Trial). AB - BACKGROUND: Co-bedding, a developmental care strategy, is the practice of caring for diaper clad twins in one incubator (versus separating and caring for each infant in separate incubators), thus creating the opportunity for skin-to-skin contact and touch between the twins. In studies of mothers and their infants, maternal skin-to-skin contact has been shown to decrease procedural pain response according to both behavioral and physiological indicators in very preterm neonates. It is uncertain if this comfort is derived solely from maternal presence or from stabilization of regulatory processes from direct skin contact. The intent of this study is to compare the comfort effect of co-bedding (between twin infants who are co-bedding and those who are not) on infant pain response and physiologic stability during a tissue breaking procedure (heelstick). METHODS/DESIGN: Medically stable preterm twin infants admitted to the Neonatal Intensive Care Unit will be randomly assigned to a co-bedding group or a standard care group. Pain response will be measured by physiological and videotaped facial reaction using the Premature Infant Pain Profile scale (PIPP). Recovery from the tissue breaking procedure will be determined by the length of time for heart rate and oxygen saturation to return to baseline. Sixty four sets of twins (n = 128) will be recruited into the study. Analysis and inference will be based on the intention-to-treat principle. DISCUSSION: If twin contact while co-bedding is determined to have a comforting effect for painful procedures, then changes in current neonatal care practices to include co-bedding may be an inexpensive, non invasive method to help maintain physiologic stability and decrease the long term psychological impact of procedural pain in this high risk population. Knowledge obtained from this study will also add to existing theoretical models with respect to the exact mechanism of comfort through touch. TRIAL REGISTRATION: NCT00917631. PMID- 20003353 TI - Diaphragmatic hernia with strangulated loop of bowel presenting after colonoscopy: case report. AB - BACKGROUND: The incidence of diaphragmatic hernias caused or exacerbated by diagnostic colonoscopy is not well elucidated at this time, and is believed to be very rare. CASE PRESENTATION: We present the case of a 57 year old man with remote history of traumatic injury who first presented with vague left shoulder pain for two weeks, mild anemia, and tested positive for fecal occult blood. Four days post colonoscopy the patient was found to have a strangulated loop of bowel herniated through the diaphragm into the left hemithorax. CONCLUSIONS: In patients with previous history of serious traumatic injury and particularly those with previous splenectomy, a thorough history and physical examination before routine colonoscopy is important. A high level of suspicion for post-operative complications should also be maintained when assessing such patients. PMID- 20003352 TI - Th1 and Th17 hypercytokinemia as early host response signature in severe pandemic influenza. AB - INTRODUCTION: Human host immune response following infection with the new variant of A/H1N1 pandemic influenza virus (nvH1N1) is poorly understood. We utilize here systemic cytokine and antibody levels in evaluating differences in early immune response in both mild and severe patients infected with nvH1N1. METHODS: We profiled 29 cytokines and chemokines and evaluated the haemagglutination inhibition activity as quantitative and qualitative measurements of host immune responses in serum obtained during the first five days after symptoms onset, in two cohorts of nvH1N1 infected patients. Severe patients required hospitalization (n = 20), due to respiratory insufficiency (10 of them were admitted to the intensive care unit), while mild patients had exclusively flu-like symptoms (n = 15). A group of healthy donors was included as control (n = 15). Differences in levels of mediators between groups were assessed by using the non parametric U Mann Whitney test. Association between variables was determined by calculating the Spearman correlation coefficient. Viral load was performed in serum by using real-time PCR targeting the neuraminidase gene. RESULTS: Increased levels of innate-immunity mediators (IP-10, MCP-1, MIP-1beta), and the absence of anti nvH1N1 antibodies, characterized the early response to nvH1N1 infection in both hospitalized and mild patients. High systemic levels of type-II interferon (IFN gamma) and also of a group of mediators involved in the development of T-helper 17 (IL-8, IL-9, IL-17, IL-6) and T-helper 1 (TNF-alpha, IL-15, IL-12p70) responses were exclusively found in hospitalized patients. IL-15, IL-12p70, IL-6 constituted a hallmark of critical illness in our study. A significant inverse association was found between IL-6, IL-8 and PaO2 in critical patients. CONCLUSIONS: While infection with the nvH1N1 induces a typical innate response in both mild and severe patients, severe disease with respiratory involvement is characterized by early secretion of Th17 and Th1 cytokines usually associated with cell mediated immunity but also commonly linked to the pathogenesis of autoimmune/inflammatory diseases. The exact role of Th1 and Th17 mediators in the evolution of nvH1N1 mild and severe disease merits further investigation as to the detrimental or beneficial role these cytokines play in severe illness. PMID- 20003354 TI - Serum levels of autoantibodies against C-reactive protein correlate with renal disease activity and response to therapy in lupus nephritis. AB - INTRODUCTION: Serum levels of C-reactive protein (CRP) seldom reflect disease activity in systemic lupus erythematosus (SLE). We have previously shown that autoantibodies against neo-epitopes of CRP often occur in SLE, but that this does not explain the modest CRP response seen in flares. However, we have repeatedly found that anti-CRP levels parallel lupus disease activity, with highest levels in patients with renal involvement; thus, we aimed to study anti-CRP in a material of well-characterized lupus nephritis patients. METHODS: Thirty-eight patients with lupus nephritis were included. Treatment with corticosteroids combined with cyclophosphamide, mycophenolate mofetil or rituximab was started after baseline kidney biopsy. A second biopsy was taken after > or = 6 months. Serum creatinine, cystatin C, complement, anti-dsDNA, anti-CRP and urinalysis were done on both occasions. Biopsies were evaluated regarding World Health Organisation (WHO) class and indices of activity and chronicity. Renal disease activity was estimated using the British Isles Lupus Assessment Group (BILAG) index. RESULTS: At baseline, 34/38 patients had renal BILAG-A; 4/38 had BILAG-B. Baseline biopsies showed WHO class III (n = 8), IV (n = 19), III to IV/V (n = 3) or V (n = 8) nephritis. Seventeen out of 38 patients were anti-CRP-positive at baseline, and six at follow-up. Overall, anti-CRP levels had dropped at follow-up (P < 0.0001) and anti-CRP levels correlated with renal BILAG (r = 0.29, P = 0.012). A positive anti-CRP test at baseline was superior to anti-dsDNA and C1q in predicting poor response to therapy as judged by renal BILAG. Baseline anti CRP levels correlated with renal biopsy activity (r = 0.33, P = 0.045), but not with chronicity index. Anti-CRP levels were positively correlated with anti-dsDNA (fluorescence-enhanced immunoassay: r = 0.63, P = 0.0003; Crithidia luciliae immunofluorescence microscopy test: r = 0.44, P < 0.0001), and inversely with C3 (r = 0.35, P = 0.007) and C4 (r = 0.29, P = 0.02), but not with C1q (r = 0.14, P = 0.24). No associations with urinary components, creatinine, cystatin C or the glomerular filtration rate were found. CONCLUSIONS: In the present study, we demonstrate a statistically significant correlation between anti-CRP levels and histopathological activity in lupus nephritis, whereas a baseline positive anti CRP test predicted poor response to therapy. Our data also confirm previous findings of associations between anti-CRP and disease activity. This indicates that anti-CRP could be helpful to assess disease activity and response to therapy in SLE nephritis, and highlights the hypothesis of a pathogenetic role for anti CRP antibodies in lupus nephritis. PMID- 20003355 TI - The transcriptional programme of Salmonella enterica serovar Typhimurium reveals a key role for tryptophan metabolism in biofilms. AB - BACKGROUND: Biofilm formation enhances the capacity of pathogenic Salmonella bacteria to survive stresses that are commonly encountered within food processing and during host infection. The persistence of Salmonella within the food chain has become a major health concern, as biofilms can serve as a reservoir for the contamination of food products. While the molecular mechanisms required for the survival of bacteria on surfaces are not fully understood, transcriptional studies of other bacteria have demonstrated that biofilm growth triggers the expression of specific sets of genes, compared with planktonic cells. Until now, most gene expression studies of Salmonella have focused on the effect of infection-relevant stressors on virulence or the comparison of mutant and wild type bacteria. However little is known about the physiological responses taking place inside a Salmonella biofilm. RESULTS: We have determined the transcriptomic and proteomic profiles of biofilms of Salmonella enterica serovar Typhimurium. We discovered that 124 detectable proteins were differentially expressed in the biofilm compared with planktonic cells, and that 10% of the S. Typhimurium genome (433 genes) showed a 2-fold or more change in the biofilm compared with planktonic cells. The genes that were significantly up-regulated implicated certain cellular processes in biofilm development including amino acid metabolism, cell motility, global regulation and tolerance to stress. We found that the most highly down-regulated genes in the biofilm were located on Salmonella Pathogenicity Island 2 (SPI2), and that a functional SPI2 secretion system regulator (ssrA) was required for S. Typhimurium biofilm formation. We identified STM0341 as a gene of unknown function that was needed for biofilm growth. Genes involved in tryptophan (trp) biosynthesis and transport were up regulated in the biofilm. Deletion of trpE led to decreased bacterial attachment and this biofilm defect was restored by exogenous tryptophan or indole. CONCLUSIONS: Biofilm growth of S. Typhimurium causes distinct changes in gene and protein expression. Our results show that aromatic amino acids make an important contribution to biofilm formation and reveal a link between SPI2 expression and surface-associated growth in S. Typhimurium. PMID- 20003356 TI - Selection of reference genes for quantitative real-time PCR in equine in vivo and fresh and frozen-thawed in vitro blastocysts. AB - BACKGROUND: Application of reverse transcription quantitative real-time polymerase chain reaction is very well suited to reveal differences in gene expression between in vivo and in vitro produced embryos. Ultimately, this may lead to optimized equine assisted reproductive techniques. However, for a correct interpretation of the real-time PCR results, all data must be normalized, which is most reliably achieved by calculating the geometric mean of the most stable reference genes. In this study a set of reliable reference genes was identified for equine in vivo and fresh and frozen-thawed in vitro embryos. FINDINGS: The expression stability of 8 candidate reference genes (ACTB, GAPDH, H2A/I, HPRT1, RPL32, SDHA, TUBA4A, UBC) was determined in 3 populations of equine blastocysts (fresh in vivo, fresh and frozen-thawed in vitro embryos). Application of geNorm indicated UBC, GAPDH, ACTB and HPRT1 as the most stable genes in the in vivo embryos and UBC, RPL32, GAPDH and ACTB in both in vitro populations. When in vivo and in vitro embryos were combined, UBC, ACTB, RPL32 and GAPDH were found to be the most stable. SDHA and H2A/I appeared to be highly regulated. CONCLUSIONS: Based on these results, the geometric mean of UBC, ACTB, RPL32 and GAPDH is to be recommended for accurate normalization of quantitative real-time PCR data in equine in vivo and in vitro produced blastocysts. PMID- 20003357 TI - Effectiveness of combined intermittent preventive treatment for children and timely home treatment for malaria control. AB - BACKGROUND: Whiles awaiting for the arrival of an effective and affordable malaria vaccine, there is a need to make use of the available control tools to reduce malaria risk, especially in children under five years and pregnant women. Intermittent preventive treatment (IPT) has recently been accepted as an important component of the malaria control strategy. This study explored the potential of a strategy of intermittent preventive treatment for children (IPTC) and timely treatment of malaria-related febrile illness in the home in reducing the parasite prevalence and malaria morbidity in young children in a coastal village in Ghana. METHODS: The study combined home-based delivery of IPTC among six to 60 months old and home treatment of suspected febrile malaria illness within 24 hours. All children between six and 60 months of age received intermittent preventive treatment using amodiaquine and artesunate, delivered by community assistants every four months (three times in 12 months). Malaria parasite prevalence surveys were conducted before the first and after the third dose of IPTC. RESULTS: Parasite prevalence was reduced from 25% to 3% (p < 0.00, Mann-Whitney) one year after the inception of the two interventions. At baseline, 13.8% of the children were febrile (axillary temperature greater than or equal to 37.5 degree Celsius) compared to 2.2% at evaluation (post IPTC3 combined with timely home management of fever) (p < 0.00, Mann-Whitney). CONCLUSION: The evaluation result indicates that IPTC given three times in a year combined with timely treatment of febrile malaria illness, impacts significantly on the parasite prevalence. The marked reduction in the parasite prevalence with this strategy points to the potential for reducing malaria-related childhood morbidity and mortality, and this should be explored by control programme managers. PMID- 20003358 TI - The impacts of substance abuse and dependence on neuropsychological functions in a sample of patients from Saudi Arabia. AB - BACKGROUND: A lot of studies were directed to explore the relation between drug abuse and neuropsychological functions. Some studies reported that even after a long duration of disappearance of withdrawal or intoxication symptoms, many patients have obvious deterioration of cognitive functions. The aim of this study was to explore the relationship between the substance use disorders and the executive functions. METHODS: Two groups were selected for this study. An experimental group consisted of 154 patients and further subdivided according to the substance used into three different subgroups: opioid, amphetamine and alcohol groups which included 49, 56 and 49 patients respectively. The control group was selected matching the experimental group in the demographic characteristics and included 100 healthy persons. Tools used were: Benton visual retention tests, color trail making test, Stroop colors-word test, symbol digit modalities test, the five dots cognitive flexibility test, and TAM verbal flexibility test. All the data were subjected to statistical analysis RESULTS: The study showed that the group of drug-dependent subjects performed significantly worse than the comparison group on all measures Also, there were significant differences among the subgroups as the alcoholic group was much worse followed by the amphetamine then the opioids groups. Patients with longer duration of dependence and multiple hospital readmissions were much worse in comparison to patients with shorter duration of dependence and less readmission. CONCLUSION: The study confirmed that the functions of specific brain regions underlying cognitive control are significantly impaired in patients of drug addiction. This impairment was significantly related to type of substance, duration of use and number of hospitalization and may contribute to most of behavioral disturbances found in addicts and need much attention during tailoring of treatment programs. PMID- 20003359 TI - Breast cancer in Kurdish women of northern Iraq: incidence, clinical stage, and case control analysis of parity and family risk. AB - BACKGROUND: Breast cancer in the Middle-East occurs in relatively young women and frequently presents as advanced disease. A protective effect of multiparity is not apparent, and high familial risk is reported in some countries. This study investigates breast cancer rates and clinical stage related to age in the Kurdish region of Iraq and evaluates risk associated with parity and family history. Findings are compared with nearby countries and the West. METHODS: Sulaimaniyah Directorate of Health records identified 539 women diagnosed with breast cancer during 2006-2008. Clinical survey forms were completed on 296 patients and on 254 age-matched controls. Age specific incidence rates were calculated from Directorate of Health population estimates. RESULTS: Average patient age was 47.4 +/- 11 years and 59.5% were pre-menopausal. Diagnosis was at clinical stage 1 for 4.1%, stage 2 for 43.5%, stage 3 for 26.0%, and stage 4 for 8.1% of patients. For 18.2%, stage was unknown. Annual breast cancer incidence rates per 100,000 women peaked at 168.9 at age 55 to 59 and declined to 57.3 at 60 and above. Patients had an average of 5.0 +/- 3.3 children compared to 5.4 +/- 3.5 for controls, P = 0.16. A first degree family member had breast cancer among 11.1% of patients and 2.1% of controls (P < 0.001) with > 50% of these patients and controls being > or =50 years old. No statistically significant relationship was found between tumor stage and age, P = 0.59. CONCLUSIONS: In Kurdish Iraq, breast cancer is predominantly a disease of pre-menopausal women having multiple pregnancies. For younger patients, breast cancer incidence was similar to the West and possibly higher than many Middle-Eastern countries, but unlike the West, the estimated rates declined markedly in the elderly. The familial breast cancer risk for both older and younger women was within the general population risk of Western countries. Clinical stages were advanced and indicated delays in diagnosis that were unrelated to patient age. PMID- 20003360 TI - Molecular approaches to the analysis of deformed wing virus replication and pathogenesis in the honey bee, Apis mellifera. AB - BACKGROUND: For years, the understanding of the pathogenetic mechanisms that underlie honey bee viral diseases has been severely hindered because of the lack of a cell culture system for virus propagation. As a result, it is very imperative to develop new methods that would permit the in vitro pathogenesis study of honey bee viruses. The identification of virus replication is an important step towards the understanding of the pathogenesis process of viruses in their respective hosts. In the present study, we developed a strand-specific RT-PCR-based method for analysis of Deformed Wing Virus (DWV) replication in honey bees and in honey bee parasitic mites, Varroa Destructor. RESULTS: The results shows that the method developed in our study allows reliable identification of the virus replication and solves the problem of falsely-primed cDNA amplifications that commonly exists in the current system. Using TaqMan real time quantitative RT-PCR incorporated with biotinylated primers and magnetic beads purification step, we characterized the replication and tissue tropism of DWV infection in honey bees. We provide evidence for DWV replication in the tissues of wings, head, thorax, legs, hemolymph, and gut of honey bees and also in Varroa mites. CONCLUSION: The strategy reported in the present study forms a model system for studying bee virus replication, pathogenesis and immunity. This study should be a significant contribution to the goal of achieving a better understanding of virus pathogenesis in honey bees and to the design of appropriate control measures for bee populations at risk to virus infections. PMID- 20003361 TI - Paraganglioma of the filum terminale: case report. AB - BACKGROUND: Paragangliomas affecting the filum terminale are extremely rare, benign tumors. The literature yielded thirty-two cases of paraganglioma in this site. CASE PRESENTATION: A 49 year-old-man, whose presenting symptoms were low back pain and left leg weakness, was diagnosed as having a paraganglioma of the filum terminale. The clinical, histological and radiological characteristics of this case, that brings the total number of cases described to 33, are discussed in the light of published data. CONCLUSIONS: This extremely rare pathology can usually be successfully treated by total surgical resection, which represents the gold standard. In the event of incomplete removal, assiduous long-term follow-up is mandatory. PMID- 20003362 TI - Environmental DNA sequencing primers for eutardigrades and bdelloid rotifers. AB - BACKGROUND: The time it takes to isolate individuals from environmental samples and then extract DNA from each individual is one of the problems with generating molecular data from meiofauna such as eutardigrades and bdelloid rotifers. The lack of consistent morphological information and the extreme abundance of these classes makes morphological identification of rare, or even common cryptic taxa a large and unwieldy task. This limits the ability to perform large-scale surveys of the diversity of these organisms.Here we demonstrate a culture-independent molecular survey approach that enables the generation of large amounts of eutardigrade and bdelloid rotifer sequence data directly from soil. Our PCR primers, specific to the 18s small-subunit rRNA gene, were developed for both eutardigrades and bdelloid rotifers. RESULTS: The developed primers successfully amplified DNA of their target organism from various soil DNA extracts. This was confirmed by both the BLAST similarity searches and phylogenetic analyses. Tardigrades showed much better phylogenetic resolution than bdelloids. Both groups of organisms exhibited varying levels of endemism. CONCLUSION: The development of clade-specific primers for characterizing eutardigrades and bdelloid rotifers from environmental samples should greatly increase our ability to characterize the composition of these taxa in environmental samples. Environmental sequencing as shown here differs from other molecular survey methods in that there is no need to pre-isolate the organisms of interest from soil in order to amplify their DNA. The DNA sequences obtained from methods that do not require culturing can be identified post-hoc and placed phylogenetically as additional closely related sequences are obtained from morphologically identified conspecifics. Our non-cultured environmental sequence based approach will be able to provide a rapid and large-scale screening of the presence, absence and diversity of Bdelloidea and Eutardigrada in a variety of soils. PMID- 20003363 TI - The expressed needs of people with chronic fatigue syndrome/myalgic encephalomyelitis: a systematic review. AB - BACKGROUND: We aimed to review systematically the needs for support in managing illness and maintaining social inclusion expressed by people with chronic fatigue syndrome/myalgic encephalomyelitis (CFS/ME) METHODS: We carried out a systematic review of primary research and personal ('own') stories expressing the needs of people with CFS/ME. Structured searches were carried out on Medline, AMED, CINAHL, EMBASE, ASSIA, CENTRAL, and other health, social and legal databases from inception to November 2007. Study inclusion, data extraction and risk of bias were assessed independently in duplicate. Expressed needs were tabulated and a conceptual framework developed through an iterative process. RESULTS: Thirty two quantitative and qualitative studies, including the views of over 2500 people with CFS/ME with mainly moderate or severe illness severity, met the inclusion criteria. The following major support needs emerged: 1) The need to make sense of symptoms and gain diagnosis, 2) for respect and empathy from service providers, 3) for positive attitudes and support from family and friends, 4) for information on CFS/ME, 5) to adjust views and priorities, 6) to develop strategies to manage impairments and activity limitations, and 7) to develop strategies to maintain/regain social participation. CONCLUSIONS: Although the studies were heterogeneous, there was consistent evidence that substantial support is needed to rebuild lives. Gaining support depends - most importantly - on the ability of providers of health and social care, colleagues, friends and relatives, and those providing educational and leisure services, to understand and respond to those needs. PMID- 20003364 TI - Cost-utility analysis of infliximab and adalimumab for refractory ulcerative colitis. AB - OBJECTIVE: To evaluate cost-utility of infliximab and adalimumab for the treatment of moderate-to-severe ulcerative colitis (UC) refractory to conventional therapies in Canada. METHODS: A Markov model was constructed to evaluate incremental cost-utility ratios (ICUR) of 5 mg/kg and 10 mg/kg infliximab and adalimumab therapies compared to 'usual care' in treating a hypothetical cohort of patients (aged 40 years and weighing 80 kg) over a five year time horizon from the perspective of a publicly-funded health care system. Clinical parameters were derived from the Active Ulcerative Colitis Trials 1 and 2. Costs were obtained through provincial drug benefit plans. ICUR was the main outcome measure and both deterministic and probabilistic sensitivity analyses were conducted. RESULTS: Compared to the strategy A ('usual care') in the base case analysis, the ICURs were CA$358,088/QALY for the strategy B ('5 mg/kg infliximab + adalimumab') and CA$575,540/QALY for the strategy C ('5 mg/kg and 10 mg/kg infliximab + adalimumab'). The results were sensitive to: the remission rates maintained in responders to 'usual care' and to 5 mg/kg infliximab, the rate of remission induced by adalimumab in non-responders to 5 mg/kg infliximab, early surgery rate, and utility values. When the willingness to pay (WTP) was less than CA$150,000/QALY, the probability of 'usual care' being the optimal strategy was 1.0. The probability of strategy B being optimal was 0.5 when the WTP approximated CA$400,000/QALY. CONCLUSIONS: The ICURs of anti-TNF-alpha drugs were not satisfactory in treating patients with moderate-to-severe refractory UC. Future research could be aimed at the long-term clinical benefits of these drugs, especially adalimumab for patients intolerant or unresponsive to infliximab treatment. PMID- 20003365 TI - Prediction of calcium-binding sites by combining loop-modeling with machine learning. AB - BACKGROUND: Protein ligand-binding sites in the apo state exhibit structural flexibility. This flexibility often frustrates methods for structure-based recognition of these sites because it leads to the absence of electron density for these critical regions, particularly when they are in surface loops. Methods for recognizing functional sites in these missing loops would be useful for recovering additional functional information. RESULTS: We report a hybrid approach for recognizing calcium-binding sites in disordered regions. Our approach combines loop modeling with a machine learning method (FEATURE) for structure-based site recognition. For validation, we compared the performance of our method on known calcium-binding sites for which there are both holo and apo structures. When loops in the apo structures are rebuilt using modeling methods, FEATURE identifies 14 out of 20 crystallographically proven calcium-binding sites. It only recognizes 7 out of 20 calcium-binding sites in the initial apo crystal structures.We applied our method to unstructured loops in proteins from SCOP families known to bind calcium in order to discover potential cryptic calcium binding sites. We built 2745 missing loops and evaluated them for potential calcium binding. We made 102 predictions of calcium-binding sites. Ten predictions are consistent with independent experimental verifications. We found indirect experimental evidence for 14 other predictions. The remaining 78 predictions are novel predictions, some with intriguing potential biological significance. In particular, we see an enrichment of beta-sheet folds with predicted calcium binding sites in the connecting loops on the surface that may be important for calcium-mediated function switches. CONCLUSION: Protein crystal structures are a potentially rich source of functional information. When loops are missing in these structures, we may be losing important information about binding sites and active sites. We have shown that limited loop modeling (e.g. loops less than 17 residues) combined with pattern matching algorithms can recover functions and propose putative conformations associated with these functions. PMID- 20003367 TI - No fate but what we make: a case of full recovery after out-of-hospital cardiac arrest. AB - An 80 years old man suffered a cardiac arrest shortly after arrival to his local health department. Basic Life Support was started promptly and nine minutes later, on evaluation by an Advanced Life Support team, the victim was defibrillated with a 200J shock. When orotracheal intubation was attempted, masseter muscle contraction was noticed: on reevaluation, the victim had pulse and spontaneous breathing.Thirty minutes later, the patient had been transferred to an emergency department. As he complained of chest pain, the ECG showed a ST segment depression in leads V4 to V6 and laboratory tests showed cardiac troponine I slightly elevated. A coronary angiography was performed urgently: significant left main plus three vessel coronary artery disease was disclosed.Eighteen hours after the cardiac arrest, a quadruple coronary artery bypass grafting operation was undertaken. During surgery, a fresh thrombus was removed from the middle left anterior descendent artery. Post-operative course was uneventful and the patient was discharged seven days after the procedure. Twenty four months later, he remains asymptomatic.In this case, the immediate call for the Advanced Life Support team, prompt basic life support and the successful defibrillation, altogether, contributed for the full recovery. Furthermore, the swiftness in the detection and treatment of the acute reversible cause (myocardial ischemia in this case) was crucial for long-term prognosis. PMID- 20003366 TI - Quantitative TaqMan real-time PCR assays for gene expression normalisation in feline tissues. AB - BACKGROUND: Gene expression analysis is an important tool in contemporary research, with real-time PCR as the method of choice for quantifying transcription levels. Co-analysis of suitable reference genes is crucial for accurate expression normalisation. Reference gene expression may vary, e.g., among species or tissues; thus, candidate genes must be tested prior to use in expression studies. The domestic cat is an important study subject in both medical research and veterinary medicine. The aim of the present study was to develop TaqMan real-time PCR assays for eight potential reference genes and to test their applicability for feline samples, including blood, lymphoid, endocrine, and gastrointestinal tissues from healthy cats, and neoplastic tissues from FeLV-infected cats. RESULTS: RNA extraction from tissues was optimised for minimal genomic DNA (gDNA) contamination without use of a DNase treatment. Real time PCR assays were established and optimised for v-abl Abelson murine leukaemia viral oncogene homolog (ABL), beta-actin (ACTB), beta-2-microglobulin (B2M), beta glucuronidase (GUSB), hydroxymethyl-bilane synthase (HMBS), hypoxanthine phosphoribosyltransferase (HPRT), ribosomal protein S7 (RPS7), and tryptophan 5 monooxygenase activation protein, zeta polypeptide (YWHAZ). The presence of pseudogenes was confirmed for four of the eight investigated genes (ACTB, HPRT, RPS7, and YWHAZ). The assays were tested together with previously developed TaqMan assays for feline glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and the universal 18S rRNA gene. Significant differences were found among the expression levels of the ten candidate reference genes, with a ~106-fold expression difference between the most abundant (18S rRNA) and the least abundant genes (ABL, GUSB, and HMBS). The expression stability determined by the geNorm and NormFinder programs differed significantly. Using the ANOVA-based NormFinder program, RPS7 was the most stable gene in the tissues studied, followed by ACTB and ABL; B2M, HPRT, and the 18S rRNA genes were the least stable ones. CONCLUSION: The reference gene expression stability varied considerably among the feline tissues investigated. No tested gene was optimal for normalisation in all tissues. For the majority of the tissues, two to three reference genes were necessary for accurate normalisation. The present study yields essential information on the correct choice of feline reference genes depending on the tissues analysed. PMID- 20003368 TI - Clinical profile, outcomes and improvement in symptoms and productivity in rhinitic patients in Karachi, Pakistan. AB - BACKGROUND: Rhinitis can cause a heavy toll on patients because of its bothersome effects on productivity. This retrospective study was conducted to explore the clinical profile, outcomes and improvement in the symptoms and productivity resulting from treatment of allergic rhinitis in Pakistan. METHODS: We carried out a retrospective file review of all allergic rhinitis patients who presented to the Ear, Nose, Throat Consulting Clinic from January, 2006 to June, 2008 using a structured proforma especially designed for this purpose. Data was entered and analyzed using SPSS v. 16.0. RESULTS: The charts of 169 patients were reviewed. The mean age of the patients was 35.2 +/- 9.1 years. Sixty percent patients were male. Ninety eight patients (58%) reported allergy symptoms to be present at both home and work. One hundred and two patients (60.4%) had symptoms severe enough to cause absence from work or academic activities. Up to seventy one percent patients were spending between 1000 - 3000 Pakistani Rupees (1 US$= 83.3 Pakistani rupees) on the treatment of allergic rhinitis per year. One hundred and fifty one patients (89.3%) reported an improvement in rhinitic symptoms and productivity while 18 patients (10.7%) didn't. This improvement was significantly associated with satisfaction with treatment (p < 0.001). CONCLUSION: Allergic rhinitis, a ubiquitous disease, was seen to cause a strain on patients in the form of recurrent treatment-related expenses as well as absenteeism from work or other daily activities. Symptoms and productivity improved significantly after treatment. PMID- 20003369 TI - An analysis of farmers' perception of the new cooperative medical system in Liaoning Province, China. AB - BACKGROUND: Since 2003, the number of pilot areas of the New Rural Cooperative Medical System (NRCMS) has increased in rural China. And the major efforts have been concentrated on the enrollment of prospective members. In this study, we examined the satisfaction of the rural residents with the NRCMS as well as factors affecting their attitudes towards the NRCMS. METHODS: The data for this study were collected from a survey involving twenty counties in Liaoning Province. Interviews and focus groups were conducted between 10th January and 20th August 2008. A total of 2,780 people aged 18-72 were randomly selected and interviewed. Data were evaluated by nonparametric tests and ordinal regression models. RESULTS: 71.6% of the study subjects were satisfied with the NRCMS. Single factor analysis showed that attitudes towards the NRCMS were influenced by gender, age, marital status, and self-rated health status. In the ordinal regression analysis, gender, age, and self-rated health status affect satisfaction (P < 0.05). CONCLUSIONS: We found that a considerable proportion of farmers were satisfied with the NRCMS. Gender, age, and self-rated health status had significant effects on farmers' attitudes towards the NRCMS. The Chinese Central Government attempted to adopt active measures in the future to continuously improve the NRCMS, including initiating educational programs, building new medical facilities and increasing financial investment. PMID- 20003370 TI - Building resilience: a preliminary exploration of women's perceptions of the use of acupuncture as an adjunct to In Vitro Fertilisation. AB - BACKGROUND: In Vitro Fertilisation (IVF) is now an accepted and effective treatment for infertility, however IVF is acknowledged as contributing to, rather than lessening, the overall psychosocial effects of infertility. Psychological and counselling interventions have previously been widely recommended in parallel with infertility treatments but whilst in many jurisdictions counselling is recommended or mandatory, it may not be widely used. Acupuncture is increasingly used as an adjunct to IVF, in this preliminary study we sought to investigate the experience of infertile women who had used acupuncture to improve their fertility. METHODS: A sample of 20 women was drawn from a cohort of women who had attended for a minimum of four acupuncture sessions in the practices of two acupuncturists in South Australia. Eight women were interviewed using a semi structured questionnaire. Six had sought acupuncture during IVF treatment and two had begun acupuncture to enhance their fertility and had later progressed to IVF. Descriptive content analysis was employed to analyse the data. RESULTS: Four major categories of perceptions about acupuncture in relation to reproductive health were identified: (a) Awareness of, and perceived benefits of acupuncture; (b) perceptions of the body and the impact of acupuncture upon it; (c) perceptions of stress and the impact of acupuncture on resilience; and (d) perceptions of the intersection of medical treatment and acupuncture. CONCLUSION: This preliminary exploration, whilst confined to a small sample of women, confirms that acupuncture is indeed perceived by infertile women to have an impact to their health. All findings outlined here are reported cautiously because they are limited by the size of the sample. They suggest that further studies of acupuncture as an adjunct to IVF should systematically explore the issues of wellbeing, anxiety, personal and social resilience and women's identity in relation to sexuality and reproduction. PMID- 20003371 TI - Short-term and long-term success of electrical cardioversion in atrial fibrillation in managed care system. AB - BACKGROUND: Initial success of electrical cardioversion (ECV) of atrial fibrillation (AF) has been reported in several studies as 50%-90%, of which only 50% patients remain in sinus rhythm (SR) at the end of one year. We conducted this study to see if outcomes of other trials are applicable in managed care setting. METHODS: We conducted a retrospective study in 370 consecutive patients who underwent ECV for AF. They were reviewed for initial outcome of ECV and recurrence of AF after a successful ECV, with and without prophylactic antiarrhythmic drugs. RESULTS: Initial success of ECV for AF was 65.7%. At one year, 47% remained in SR. AF for Val). Immunohistochemistry staining was used to measure the level of Aurora-A expression in bladder and colon cancer specimens. To reveal the effect of overexpression of the above two genes on cellular responses, mouse NIH3T3 fibroblast derived cell lines over-expressing either RasV12 and wild-type Aurora A (designated WT) or RasV12 and kinase-inactivated Aurora-A (KD) were established. MTT and focus formation assays were conducted to measure proliferation rate and focus formation capability of the cells. Small interfering RNA, pharmacological inhibitors and dominant negative genes were used to dissect the signaling pathways involved. RESULTS: Overexpression of wild-type Aurora-A and mutation of RasV12 were detected in human bladder and colon cancer tissues. Wild-type Aurora-A induces focus formation and aggregation of the RasV12 transformants. Aurora-A activates Ral A and the phosphorylation of AKT as well as enhances the phosphorylation of MEK, ERK of WT cells. Finally, the Ras/MEK/ERK signaling pathway is responsible for Aurora-A induced aggregation of the RasV12 transformants. CONCLUSION: Wild-type-Aurora-A enhances focus formation and aggregation of the RasV12 transformants and the latter occurs through modulating the Ras/MEK/ERK signaling pathway. PMID- 20003377 TI - Association of HLA-A, B, DRB1 alleles and haplotypes with HIV-1 infection in Chongqing, China. AB - BACKGROUND: The human immunodeficiency virus type 1(HIV-1) epidemic in Chongqing, China, is increasing rapidly with the dominant subtype of CRF07_BC over the past 3 years. Since human leukocyte antigen (HLA) polymorphisms have shown strong association with susceptibility/resistance to HIV-1 infection from individuals with different ethnic backgrounds, a recent investigation on frequencies of HLA class I and class II alleles in a Chinese cohort also indicated that similar correlation existed in HIV infected individuals from several provinces in China, however, such information is unavailable in Chongqing, southwest China. METHODS: In this population-based study, we performed polymerase chain reaction analysis with sequence-specific oligonucleotide probes (PCR-SSOP) for intermediate-low resolution HLA typing in a cohort of 549 HIV-1 infected individuals, another 2475 healthy subjects from the Han nationality in Chongqing, China, were selected as population control. We compared frequencies of HLA-A, B, DRB1 alleles, haplotypes and genotypes between the two groups, and analyzed their association with HIV-1 susceptibility or resistance. RESULTS: The genetic profile of HLA (A, B, DRB1) alleles of HIV-1 infected individuals from Chongqing Han of China was obtained. Several alleles of HLA-B such as B*46 (P = 0.001, OR = 1.38, 95%CI = 1.13-1.68), B*1501G(B62) (P = 0.013, OR = 1.42, 95%CI = 1.08-1.88), B*67 (P = 0.022, OR = 2.76, 95%CI = 1.16-6.57), B*37 (P = 0.014, OR = 1.93, 95%CI = 1.14-3.28) and B*52 (P = 0.038, OR = 1.64, 95%CI = 1.03-2.61) were observed to have association with susceptibility to HIV-1 infection in this population. In addition, the haplotype analysis revealed that A*11-B*46, A*24-B*54 and A*01-B*37 for 2-locus, and A*11 B*46-DRB1*09, A*02-B*46-DRB1*08, A*11-B*4001G-DRB1*15, A*02-B*4001G-DRB1*04, A*11 B*46-DRB1*08 and A*02-B*4001G-DRB1*12 for 3-locus had significantly overrepresented in HIV-1 infected individuals, whereas A*11-B*1502G, A*11-B*1502G DRB1*12 and A*33-B*58-DRB1*13 were underrepresented. However, the low-resolution homozygosity of HLA-A, B, DRB1 loci and HLA-Bw4/Bw6 genotypes did not differ significantly between the two groups. CONCLUSION: These results may contribute to the database of HLA profiles in HIV-1 infected Chinese population, consequently, the association of certain HLA alleles with susceptibility or resistance to HIV-1 infection would provide with clues in choosing proper preventive strategies against HIV-1 infection and developing effective HIV-1 vaccines in Chinese population, especially for those in southwest China. PMID- 20003378 TI - Evaluation of the Palutop+4 malaria rapid diagnostic test in a non-endemic setting. AB - BACKGROUND: Palutop+4 (All. Diag, Strasbourg, France), a four-band malaria rapid diagnostic test (malaria RDT) targeting the histidine-rich protein 2 (HRP-2), Plasmodium vivax-specific parasite lactate dehydrogenase (Pv-pLDH) and pan Plasmodium-specific pLDH (pan-pLDH) was evaluated in a non-endemic setting on stored whole blood samples from international travellers suspected of malaria. METHODS: Microscopy corrected by PCR was the reference method. Samples include those infected by Plasmodium falciparum (n = 323), Plasmodium vivax (n = 97), Plasmodium ovale (n = 73) and Plasmodium malariae (n = 25) and 95 malaria negative samples. RESULTS: The sensitivities for the diagnosis of P. falciparum, P. vivax, P. malariae and P. ovale were 85.1%, 66.0%, 32.0% and 5.5%. Sensitivities increased at higher parasite densities and reached 90.0% for P. falciparum >100/microl and 83.8% for P. vivax > 500/microl. Fourteen P. falciparum samples reacted with the Pv-pLDH line, one P. vivax sample with the HRP-2 line, and respectively two and four P. ovale and P. malariae samples reacted with the HRP-2 line. Two negative samples gave a signal with the HRP-2 line. Faint and weak line intensities were observed for 129/289 (44.6%) HRP-2 lines in P. falciparum samples, for 50/64 (78.1%) Pv-pLDH lines in P. vivax samples and for 9/13 (69.2%) pan-pLDH lines in P. ovale and P. malariae samples combined. Inter-observer reliabilities for positive and negative readings were excellent for the HRP-2 and Pv-pLDH lines (overall agreement > 92.0% and kappa values for each pair of readers >or= 0.88), and good for the pan-pLDH line (85.5% overall agreement and kappa-values >or= 0.74). CONCLUSIONS: Palutop+4 performed moderately for the detection of P. falciparum and P. vivax, but sensitivities were lower than those of three-band malaria RDTs. PMID- 20003379 TI - A role of periaqueductal grey NR2B-containing NMDA receptor in mediating persistent inflammatory pain. AB - The midbrain periaqueductal grey (PAG) is a structure known for its roles in pain transmission and modulation. Noxious stimuli potentiate the glutamate synaptic transmission and enhance glutamate NMDA receptor expression in the PAG. However, little is known about roles of NMDA receptor subunits in the PAG in processing the persistent inflammatory pain. The present study was undertaken to investigate NR2A- and NR2B-containing NMDA receptors in the PAG and their modulation to the peripheral painful inflammation. Noxious stimuli induced by hind-paw injection of complete Freund's adjuvant (CFA) caused up-regulation of NR2B-containing NMDA receptors in the PAG, while NR2A-containing NMDA receptors were not altered. Whole-cell patch-clamp recordings revealed that NMDA receptor mediated mEPSCs were increased significantly in the PAG synapse during the chronic phases of inflammatory pain in mice. PAG local infusion of Ro 25-6981, an NR2B antagonist, notably prolonged the paw withdrawal latency to thermal radian heat stimuli bilaterally in rats. Hyperoside (Hyp), one of the flavonoids compound isolated from Rhododendron ponticum L., significantly reversed up-regulation of NR2B containing NMDA receptors in the PAG and exhibited analgesic activities against persistent inflammatory stimuli in mice. Our findings provide strong evidence that up-regulation of NR2B-containing NMDA receptors in the PAG involves in the modulation to the peripheral persistent inflammatory pain. PMID- 20003380 TI - Monte Carlo vs. pencil beam based optimization of stereotactic lung IMRT. AB - BACKGROUND: The purpose of the present study is to compare finite size pencil beam (fsPB) and Monte Carlo (MC) based optimization of lung intensity-modulated stereotactic radiotherapy (lung IMSRT). MATERIALS AND METHODS: A fsPB and a MC algorithm as implemented in a biological IMRT planning system were validated by film measurements in a static lung phantom. Then, they were applied for static lung IMSRT planning based on three different geometrical patient models (one phase static CT, density overwrite one phase static CT, average CT) of the same patient. Both 6 and 15 MV beam energies were used. The resulting treatment plans were compared by how well they fulfilled the prescribed optimization constraints both for the dose distributions calculated on the static patient models and for the accumulated dose, recalculated with MC on each of 8 CTs of a 4DCT set. RESULTS: In the phantom measurements, the MC dose engine showed discrepancies < 2%, while the fsPB dose engine showed discrepancies of up to 8% in the presence of lateral electron disequilibrium in the target. In the patient plan optimization, this translates into violations of organ at risk constraints and unpredictable target doses for the fsPB optimized plans. For the 4D MC recalculated dose distribution, MC optimized plans always underestimate the target doses, but the organ at risk doses were comparable. The results depend on the static patient model, and the smallest discrepancy was found for the MC optimized plan on the density overwrite one phase static CT model. CONCLUSIONS: It is feasible to employ the MC dose engine for optimization of lung IMSRT and the plans are superior to fsPB. Use of static patient models introduces a bias in the MC dose distribution compared to the 4D MC recalculated dose, but this bias is predictable and therefore MC based optimization on static patient models is considered safe. PMID- 20003381 TI - Modeling trends of health and health related indicators in Ethiopia (1995-2008): a time-series study. AB - BACKGROUND: The Federal Ministry of Health of Ethiopia has been publishing Health and Health related indicators of the country annually since 1987 E.C. These indicators have been of high importance in indicating the status of health in the country in those years. However, the trends/patterns of these indicators and the factors related to the trends have not yet been investigated in a systematic manner. In addition, there were minimal efforts to develop a model for predicting future values of Health and Health related indicators based on the current trend. OBJECTIVES: The overall aim of this study was to analyze trends of and develop model for prediction of Health and Health related indicators. More specifically, it described the trends of Health and Health related indicators, identified determinants of mortality and morbidity indicators and developed model for predicting future values of MDG indicators. METHODS: This study was conducted on Health and Health related indicators of Ethiopia from the year 1987 E.C to 2000 E.C. Key indicators of Mortality and Morbidity, Health service coverage, Health systems resources, Demographic and socio-economic, and Risk factor indicators were extracted and analyzed. The trends in these indicators were established using trend analysis techniques. The determinants of the established trends were identified using ARIMA models in STATA. The trend-line equations were then used to predict future values of the indicators. RESULTS: Among the mortality indicators considered in this study, it was only Maternal Mortality Ratio that showed statistically significant decrement within the study period. The trends of Total Fertility Rate, physician per 100,000 population, skilled birth attendance and postnatal care coverage were found to have significant association with Maternal Mortality Ratio trend. There was a reversal of malaria parasite prevalence in 1999 E.C from Plasmodium Falciparum to Plasmodium Vivax. Based on the prediction from the current trend, the Millennium Development Goal target for under-five mortality rate and proportion of people having access to basic sanitation can be achieved. CONCLUSION: The current trend indicates the need to accelerate the progress of the indicators to achieve MDGs at or before 2015, particularly for Maternal Health and access to safe water supply. PMID- 20003382 TI - The impact of measurement errors in the identification of regulatory networks. AB - BACKGROUND: There are several studies in the literature depicting measurement error in gene expression data and also, several others about regulatory network models. However, only a little fraction describes a combination of measurement error in mathematical regulatory networks and shows how to identify these networks under different rates of noise. RESULTS: This article investigates the effects of measurement error on the estimation of the parameters in regulatory networks. Simulation studies indicate that, in both time series (dependent) and non-time series (independent) data, the measurement error strongly affects the estimated parameters of the regulatory network models, biasing them as predicted by the theory. Moreover, when testing the parameters of the regulatory network models, p-values computed by ignoring the measurement error are not reliable, since the rate of false positives are not controlled under the null hypothesis. In order to overcome these problems, we present an improved version of the Ordinary Least Square estimator in independent (regression models) and dependent (autoregressive models) data when the variables are subject to noises. Moreover, measurement error estimation procedures for microarrays are also described. Simulation results also show that both corrected methods perform better than the standard ones (i.e., ignoring measurement error). The proposed methodologies are illustrated using microarray data from lung cancer patients and mouse liver time series data. CONCLUSIONS: Measurement error dangerously affects the identification of regulatory network models, thus, they must be reduced or taken into account in order to avoid erroneous conclusions. This could be one of the reasons for high biological false positive rates identified in actual regulatory network models. PMID- 20003384 TI - Research into the efficacy and cost-effectiveness of brief, free of charge and anonymous sex counselling to improve (mental) health in youth: design of a randomised controlled trial. AB - BACKGROUND: The capacity to form romantic relationships and sexual health of adolescents in the Netherlands are compromised by several factors, including young age of first intercourse and adolescent depression. Several thresholds like own expenses, trust and embarrassment prevent adolescents to seek help for their sexual problems. To overcome these thresholds, brief sex counselling has been developed. It has been used since 2006 within the Rotterdam-Rijnmond Public Health Service, but there is lack of information about the (cost-) effectiveness. In the current study we will evaluate the (cost-) effectiveness of brief sex counselling for sexual problems in adolescents and young adults between 18 and 25 years of age. METHODS: In a randomised controlled trial we will compare (1) brief sex counselling with (2) intensive sexological treatment, and (3) delayed treatment (waiting list). Embedded in this RCT will be a trial-based economic evaluation, looking at the cost-effectiveness and cost-utility of brief sex counselling versus the two other interventions. Four hundred fifty adolescents (aged 18-25) with sexual problems will be recruited among the persons who visit the Public Health Service (PHS) and through various websites. After a screening procedure, eligible participants will be randomly allocated to one of the three intervention groups. Primary outcome measure of the clinical evaluation is the severity of sexual problems. Other outcomes include psychological distress, especially depression. The economic evaluation will be performed from a societal perspective. Costs will be assessed continuously by a retrospective questionnaire covering the last 3 month. All outcome assessments (including those for the economic evaluation) will take place via the internet at baseline, and at 3, 6, 9, and 12 months after baseline. DISCUSSION: The proposed research project will be the first study to provide preliminary data about the effect and cost effectiveness of brief sex counselling in youth in comparison with intensive sexological treatment and delayed treatment. It is anticipated that positive results in (cost-) effectiveness of the proposed intervention will contribute to the improvement of sexual health care for adolescents and young adults. TRIAL REGISTRATION: The study has been registered at the Netherlands Trial Register, part of the Dutch Cochrane Centre (NTR1952). PMID- 20003383 TI - A systematic review of the reporting of Data Monitoring Committees' roles, interim analysis and early termination in pediatric clinical trials. AB - BACKGROUND: Decisions about interim analysis and early stopping of clinical trials, as based on recommendations of Data Monitoring Committees (DMCs), have far reaching consequences for the scientific validity and clinical impact of a trial. Our aim was to evaluate the frequency and quality of the reporting on DMC composition and roles, interim analysis and early termination in pediatric trials. METHODS: We conducted a systematic review of randomized controlled clinical trials published from 2005 to 2007 in a sample of four general and four pediatric journals. We used full-text databases to identify trials which reported on DMCs, interim analysis or early termination, and included children or adolescents. Information was extracted on general trial characteristics, risk of bias, and a set of parameters regarding DMC composition and roles, interim analysis and early termination. RESULTS: 110 of the 648 pediatric trials in this sample (17%) reported on DMC or interim analysis or early stopping, and were included; 68 from general and 42 from pediatric journals. The presence of DMCs was reported in 89 of the 110 included trials (81%); 62 papers, including 46 of the 89 that reported on DMCs (52%), also presented information about interim analysis. No paper adequately reported all DMC parameters, and nine (15%) reported all interim analysis details. Of 32 trials which terminated early, 22 (69%) did not report predefined stopping guidelines and 15 (47%) did not provide information on statistical monitoring methods. CONCLUSIONS: Reporting on DMC composition and roles, on interim analysis results and on early termination of pediatric trials is incomplete and heterogeneous. We propose a minimal set of reporting parameters that will allow the reader to assess the validity of trial results. PMID- 20003385 TI - The role of mTOR and phospho-p70S6K in pathogenesis and progression of gastric carcinomas: an immunohistochemical study on tissue microarray. AB - BACKGROUND: mTOR signaling pathway and its downstream serine/threonine kinase p70S6k were frequently activated in human cancers. The dysregulation of the mTOR pathway has been found to be a contributing factor of a variety of different cancer. To investigate the role of mTOR signal pathway in the stepwise development of gastric carcinomas, we analyzed the correlations between the mTOR and P70S6K expression and clinic pathological factors and studied its prognostic role in gastric carcinomas. METHODS: mTOR and phospho-p70S6K proteins were examined by immunohistochemistry on tissue microarray containing gastric carcinomas (n = 412), adenomas (n = 47) and non-neoplastic mucosa (NNM, n = 197) with a comparison of their expression with clinicopathological parameters of carcinomas. RESULTS: There was no difference of mTOR expression between these three tissues (p > 0.05). Cytoplasmic phospho(p)-P706SK was highly expressed in adenoma, compared with ANNMs (p < 0.05), whereas its nuclear expression was lower in gastric carcinomas than gastric adenoma and ANNMs (p < 0.05). These three markers were preferably expressed in the older patients with gastric cancer and intestinal-type carcinoma (p < 0.05). mTOR expression was positively correlated with the cytoplasmic and nuclear expression of p-P70S6K(p < 0.05). Nuclear P70S6K was inversely linked to tumor size, depth of invasion, lymph node metastasis and UICC staging (p < 0.05). Univariate analysis indicated that expression of mTOR and nuclear p-P70S6K was closely linked to favorable prognosis of the carcinoma patients (p < 0.05). Multivariate analysis showed that age, depth of invasion, lymphatic invasion, lymph node metastasis, Lauren's classification and mTOR expression were independent prognostic factors for overall gastric carcinomas (p < 0.05). CONCLUSION: Aberrant expression of p-P70S6K possibly contributes to pathogenesis, growth, invasion and metastasis of gastric carcinomas. It was considered as a promising marker to indicate the aggressive behaviors and prognosis of gastric carcinomas. PMID- 20003386 TI - Survival prediction from clinico-genomic models--a comparative study. AB - BACKGROUND: Survival prediction from high-dimensional genomic data is an active field in today's medical research. Most of the proposed prediction methods make use of genomic data alone without considering established clinical covariates that often are available and known to have predictive value. Recent studies suggest that combining clinical and genomic information may improve predictions, but there is a lack of systematic studies on the topic. Also, for the widely used Cox regression model, it is not obvious how to handle such combined models. RESULTS: We propose a way to combine classical clinical covariates with genomic data in a clinico-genomic prediction model based on the Cox regression model. The prediction model is obtained by a simultaneous use of both types of covariates, but applying dimension reduction only to the high-dimensional genomic variables. We describe how this can be done for seven well-known prediction methods: variable selection, unsupervised and supervised principal components regression and partial least squares regression, ridge regression, and the lasso. We further perform a systematic comparison of the performance of prediction models using clinical covariates only, genomic data only, or a combination of the two. The comparison is done using three survival data sets containing both clinical information and microarray gene expression data. Matlab code for the clinico genomic prediction methods is available at http://www.med.uio.no/imb/stat/bmms/software/clinico-genomic/. CONCLUSIONS: Based on our three data sets, the comparison shows that established clinical covariates will often lead to better predictions than what can be obtained from genomic data alone. In the cases where the genomic models are better than the clinical, ridge regression is used for dimension reduction. We also find that the clinico-genomic models tend to outperform the models based on only genomic data. Further, clinico genomic models and the use of ridge regression gives for all three data sets better predictions than models based on the clinical covariates alone. PMID- 20003387 TI - Dissecting the dynamics of dysregulation of cellular processes in mouse mammary gland tumor. AB - BACKGROUND: Elucidating the sequence of molecular events underlying breast cancer formation is of enormous value for understanding this disease and for design of an effective treatment. Gene expression measurements have enabled the study of transcriptome-wide changes involved in tumorigenesis. This usually occurs through identification of differentially expressed genes or pathways. RESULTS: We propose a novel approach that is able to delineate new cancer-related cellular processes and the nature of their involvement in tumorigenesis. First, we define modules as densely interconnected and functionally enriched areas of a Protein Interaction Network. Second, 'differential expression' and 'differential co-expression' analyses are applied to the genes in these network modules, allowing for identification of processes that are up- or down-regulated, as well as processes disrupted (low co-expression) or invoked (high co-expression) in different tumor stages. Finally, we propose a strategy to identify regulatory miRNAs potentially responsible for the observed changes in module activities. We demonstrate the potential of this analysis on expression data from a mouse model of mammary gland tumor, monitored over three stages of tumorigenesis. Network modules enriched in adhesion and metabolic processes were found to be inactivated in tumor cells through the combination of dysregulation and down-regulation, whereas the activation of the integrin complex and immune system response modules is achieved through increased co-regulation and up-regulation. Additionally, we confirmed a known miRNA involved in mammary gland tumorigenesis, and present several new candidates for this function. CONCLUSIONS: Understanding complex diseases requires studying them by integrative approaches that combine data sources and different analysis methods. The integration of methods and data sources proposed here yields a sensitive tool, able to pinpoint new processes with a role in cancer, dissect modulation of their activity and detect the varying assignments of genes to functional modules over the course of a disease. PMID- 20003388 TI - Modular prediction of protein structural classes from sequences of twilight-zone identity with predicting sequences. AB - BACKGROUND: Knowledge of structural class is used by numerous methods for identification of structural/functional characteristics of proteins and could be used for the detection of remote homologues, particularly for chains that share twilight-zone similarity. In contrast to existing sequence-based structural class predictors, which target four major classes and which are designed for high identity sequences, we predict seven classes from sequences that share twilight zone identity with the training sequences. RESULTS: The proposed MODular Approach to Structural class prediction (MODAS) method is unique as it allows for selection of any subset of the classes. MODAS is also the first to utilize a novel, custom-built feature-based sequence representation that combines evolutionary profiles and predicted secondary structure. The features quantify information relevant to the definition of the classes including conservation of residues and arrangement and number of helix/strand segments. Our comprehensive design considers 8 feature selection methods and 4 classifiers to develop Support Vector Machine-based classifiers that are tailored for each of the seven classes. Tests on 5 twilight-zone and 1 high-similarity benchmark datasets and comparison with over two dozens of modern competing predictors show that MODAS provides the best overall accuracy that ranges between 80% and 96.7% (83.5% for the twilight zone datasets), depending on the dataset. This translates into 19% and 8% error rate reduction when compared against the best performing competing method on two largest datasets. The proposed predictor provides accurate predictions at 58% accuracy for membrane proteins class, which is not considered by majority of existing methods, in spite that this class accounts for only 2% of the data. Our predictive model is analyzed to demonstrate how and why the input features are associated with the corresponding classes. CONCLUSIONS: The improved predictions stem from the novel features that express collocation of the secondary structure segments in the protein sequence and that combine evolutionary and secondary structure information. Our work demonstrates that conservation and arrangement of the secondary structure segments predicted along the protein chain can successfully predict structural classes which are defined based on the spatial arrangement of the secondary structures. A web server is available at http://biomine.ece.ualberta.ca/MODAS/. PMID- 20003390 TI - Rapid detection of SMARCB1 sequence variation using high resolution melting. AB - BACKGROUND: Rhabdoid tumors are rare cancers of early childhood arising in the kidney, central nervous system and other organs. The majority are caused by somatic inactivating mutations or deletions affecting the tumor suppressor locus SMARCB1 [OMIM 601607]. Germ-line SMARCB1 inactivation has been reported in association with rhabdoid tumor, epitheloid sarcoma and familial schwannomatosis, underscoring the importance of accurate mutation screening to ascertain recurrence and transmission risks. We describe a rapid and sensitive diagnostic screening method, using high resolution melting (HRM), for detecting sequence variations in SMARCB1. METHODS: Amplicons, encompassing the nine coding exons of SMARCB1, flanking splice site sequences and the 5' and 3' UTR, were screened by both HRM and direct DNA sequencing to establish the reliability of HRM as a primary mutation screening tool. Reaction conditions were optimized with commercially available HRM mixes. RESULTS: The false negative rate for detecting sequence variants by HRM in our sample series was zero. Nine amplicons out of a total of 140 (6.4%) showed variant melt profiles that were subsequently shown to be false positive. Overall nine distinct pathogenic SMARCB1 mutations were identified in a total of 19 possible rhabdoid tumors. Two tumors had two distinct mutations and two harbored SMARCB1 deletion. Other mutations were nonsense or frame-shifts. The detection sensitivity of the HRM screening method was influenced by both sequence context and specific nucleotide change and varied from 1: 4 to 1:1000 (variant to wild-type DNA). A novel method involving digital HRM, followed by re-sequencing, was used to confirm mutations in tumor specimens containing associated normal tissue. CONCLUSIONS: This is the first report describing SMARCB1 mutation screening using HRM. HRM is a rapid, sensitive and inexpensive screening technology that is likely to be widely adopted in diagnostic laboratories to facilitate whole gene mutation screening. PMID- 20003389 TI - c.1810C>T polymorphism of NTRK1 gene is associated with reduced survival in neuroblastoma patients. AB - BACKGROUND: TrkA (encoded by NTRK1 gene), the high-affinity tyrosine kinase receptor for neurotrophins, is involved in neural crest cell differentiation. Its expression has been reported to be associated with a favourable prognosis in neuroblastoma. Therefore, the entire coding sequence of NTRK1 gene has been analysed in order to identify mutations and/or polymorphisms which may alter TrkA receptor expression. METHODS: DNA was extracted from neuroblastomas of 55 Polish and 114 Italian patients and from peripheral blood leukocytes of 158 healthy controls. Denaturing High-Performance Liquid Chromatography (DHPLC) and Single Strand Conformation Polymorphism (SSCP) analysis were used to screen for sequence variants. Genetic changes were confirmed by direct sequencing and correlated with biological and clinical data. RESULTS: Three previously reported and nine new single nucleotide polymorphisms were detected. c.1810C>T polymorphism present in 8.7% of cases was found to be an independent marker of disease recurrence (OR = 13.3; p = 0.009) associated with lower survival rates (HR = 4.45 p = 0.041). c.1810C>T polymorphism's unfavourable prognostic value was most significant in patients under 18 months of age with no MYCN amplification (HR = 26; p = 0.008). In-silico analysis of the c.1810C>T polymorphism suggests that the substitution of the corresponding amino acid residue within the conservative region of the tyrosine kinase domain might theoretically interfere with the functioning of the TrkA protein. CONCLUSIONS: NTRK1 c.1810C>T polymorphism appears to be a new independent prognostic factor of poor outcome in neuroblastoma, especially in children under 18 months of age with no MYCN amplification. PMID- 20003391 TI - ERCC2, ERCC1 polymorphisms and haplotypes, cooking oil fume and lung adenocarcinoma risk in Chinese non-smoking females. AB - BACKGROUND: Excision repair cross-complementing group 1 (ERCC1) and group 2 (ERCC2) proteins play important roles in the repair of DNA damage and adducts. Single nucleotide polymorphisms (SNPs) of DNA repair genes are suspected to influence the risk of lung cancer. This study aimed to investigate the association between the ERCC2 751, 312 and ERCC1 118 polymorphisms and the risk of lung adenocarcinoma in Chinese non-smoking females. METHODS: A hospital-based case-control study of 285 patients and 285 matched controls was conducted. Information concerning demographic and risk factors was obtained for each case and control by a trained interviewer. After informed consent was obtained, each person donated 10 ml blood for biomarker testing. Three polymorphisms were determined by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method. RESULTS: This study showed that the individuals with the combined ERCC2 751AC/CC genotypes were at an increased risk for lung adenocarcinoma compared with those carrying the AA genotype [adjusted odds ratios (OR) 1.64, 95% confidence interval (CI) 1.06-2.52]. The stratified analysis suggested that increased risk associated with ERCC2 751 variant genotypes (AC/CC) was more pronounced in individuals without exposure to cooking oil fume (OR 1.98, 95%CI 1.18-3.32) and those without exposure to fuel smoke (OR 2.47, 95%CI 1.46 4.18). Haplotype analysis showed that the A-G-T and C-G-C haplotypes were associated with increased risk of lung adenocarcinoma among non-smoking females (ORs were 1.43 and 2.28, 95%CIs were 1.07-1.91 and 1.34-3.89, respectively). CONCLUSION: ERCC2 751 polymorphism may be a genetic risk modifier for lung adenocarcinoma in non-smoking females in China. PMID- 20003392 TI - An improved phage-display panning method to produce an HM-1 killer toxin anti idiotypic antibody. AB - BACKGROUND: Phage-display panning is an integral part of biomedical research. Regular panning methods are sometimes complicated by inefficient detachment of the captured phages from the antigen-coated solid supports, which prompted us to modify. Here, we produce an efficient antigen-specific single chain fragment variable (scFv) antibody by using a target-related molecule that favored selection of recombinant antibodies. RESULTS: To produce more selective and specific anti-idiotypic scFv-antibodies from a cDNA library, constructed from HM 1 killer toxin (HM-1)-neutralizing monoclonal antibodies (nmAb-KT), the method was modified by using an elution buffer supplemented with HM-1 that shares structural and functional similarities with the active site of the scFv antibody. Competitive binding of HM-1 to nmAb-KT allowed easy and quick dissociation of scFv-displayed phages from immobilized nmAb-KT to select specific anti-idiotypic scFv antibodies of HM-1. After modified panning, 80% clones (40/50) showed several times higher binding affinity to nmAb-KT than regular panning. The major populations (48%) of these clones (scFv K1) were genotypically same and had strong cytocidal activity against Saccharomyces and Candida species. The scFv K1 (K(d) value = 4.62 x 10(-8) M) had strong reactivity toward nmAb-KT, like HM-1 (K(d) value = 6.74 x 10(-9) M) as judged by SPR analysis. CONCLUSION: The scFv antibodies generated after modified subtractive panning appear to have superior binding properties and cytocidal activity than regular panning. A simple modification of the elution condition in the phage-display panning protocol makes a large difference in determining success. Our method offers an attractive platform to discover potential therapeutic candidates. PMID- 20003393 TI - TIM-Finder: a new method for identifying TIM-barrel proteins. AB - BACKGROUND: The triosephosphate isomerase (TIM)-barrel fold occurs frequently in the proteomes of different organisms, and the known TIM-barrel proteins have been found to play diverse functional roles. To accelerate the exploration of the sequence-structure protein landscape in the TIM-barrel fold, a computational tool that allows sensitive detection of TIM-barrel proteins is required. RESULTS: To develop a new TIM-barrel protein identification method in this work, we consider three descriptors: a sequence-alignment-based descriptor using PSI-BLAST e-values and bit scores, a descriptor based on secondary structure element alignment (SSEA), and a descriptor based on the occurrence of PROSITE functional motifs. With the assistance of Support Vector Machine (SVM), the three descriptors were combined to obtain a new method with improved performance, which we call TIM Finder. When tested on the whole proteome of Bacillus subtilis, TIM-Finder is able to detect 194 TIM-barrel proteins at a 99% confidence level, outperforming the PSI-BLAST search as well as one existing fold recognition method. CONCLUSIONS: TIM-Finder can serve as a competitive tool for proteome-wide TIM barrel protein identification. The TIM-Finder web server is freely accessible at http://202.112.170.199/TIM-Finder/. PMID- 20003394 TI - Ghrelin-like peptide with fatty acid modification and O-glycosylation in the red stingray, Dasyatis akajei. AB - BACKGROUND: Ghrelin (GRLN) is now known to be an appetite-stimulating and growth hormone (GH)-releasing peptide that is predominantly synthesized and secreted from the stomachs of various vertebrate species from fish to mammals. Here, we report a GRLN-like peptide (GRLN-LP) in a cartilaginous fish, the red stingray, Dasyatis akajei. RESULTS: The purified peptide contains 16 amino acids (GVSFHPQPRS10TSKPSA), and the serine residue at position 3 is modified by n octanoic acid. The modification is the characteristic of GRLN. The six N-terminal amino acid residues (GVSFHP) were identical to another elasmobranch shark GRLN-LP that was recently identified although it had low identity with other GRLN peptides. Therefore, we designated this peptide stingray GRLN-LP. Uniquely, stingray GRLN-LP was O-glycosylated with mucin-type glycan chains [N-acetyl hexosamine (HexNAc)3 hexose(Hex)2] at threonine at position 11 (Thr-11) or both serine at position 10 (Ser-10) and Thr-11. Removal of the glycan structure by O glycanase made the in vitro activity of stingray GRLN-LP decreased when it was evaluated by the increase in intracellular Ca2+ concentrations using a rat GHS R1a-expressing cell line, suggesting that the glycan structure plays an important role for maintaining the activity of stingray GRLN-LP. CONCLUSIONS: This study reveals the structural diversity of GRLN and GRLN-LP in vertebrates. PMID- 20003395 TI - Proteomic analysis during larval development and metamorphosis of the spionid polychaete Pseudopolydora vexillosa. AB - BACKGROUND: While the larval-juvenile transition (metamorphosis) in the spionid polychaete Pseudopolydora vexillosa involves gradual morphological changes and does not require substantial development of juvenile organs, the opposite occurs in the barnacle Balanus amphitrite. We hypothesized that the proteome changes during metamorphosis in the spionids are less drastic than that in the barnacles. To test this, proteomes of pre-competent larvae, competent larvae (ready to metamorphose), and juveniles of P. vexillosa were compared using 2-dimensional gel electrophoresis (2-DE), and they were then compared to those of the barnacle. RESULTS: Unlike the significant changes found during barnacle metamorphosis, proteomes of competent P. vexillosa larvae were more similar to those of their juveniles. Pre-competent larvae had significantly fewer protein spots (384 spots), while both competent larvae and juveniles expressed about 660 protein spots each. Proteins up-regulated during competence identified by MALDI-TOF/TOF analysis included a molecular chaperon (calreticulin), a signal transduction regulator (tyrosin activation protein), and a tissue-remodeling enzyme (metallopeptidase). CONCLUSIONS: This was the first time to study the protein expression patterns during the metamorphosis of a marine polychaete and to compare the proteomes of marine invertebrates that have different levels of morphological changes during metamorphosis. The findings provide promising initial steps towards the development of a proteome database for marine invertebrate metamorphosis, thus deciphering the possible mechanisms underlying larval metamorphosis in non-model marine organisms. PMID- 20003396 TI - A comparative study of a flow-cytometry-based assessment of in vitro Plasmodium falciparum drug sensitivity. AB - BACKGROUND: Recently developed Sybr Green-based in vitro Plasmodium falciparum drug sensitivity assays provide an attractive alternative to current manual and automated methods. The present study evaluated flow cytometry measurement of DNA staining with Sybr Green in comparison with the P. falciparum lactate dehydrogenase assay, the tritiated hypoxanthine incorporation assay, a previously described Sybr Green based plate reader assay and light microscopy. METHODS: All assays were set up in standardized format in 96-well plates. The 50% inhibitory concentrations (IC50) of chloroquine, mefloquine and dihydroartemisinin against the laboratory adapted P. falciparum strains 3D7, E8B, W2mef and Dd2 were determined using each method. RESULTS: The resolution achieved by flow cytometry allowed quantification of the increase in individual cell DNA content after an incubation period of only 24 h. Regression, and Bland and Altman analyses showed that the IC50 values determined using the flow cytometry assay after 24 h agreed well with those obtained using the hypoxanthine incorporation assay, the P. falciparum lactate dehydrogenase assay, the Sybr Green plate reader assay and light microscopy. However the values obtained with the flow cytometry assay after 48 h of incubation differed significantly from those obtained with the hypoxanthine incorporation assay, and the P. falciparum lactate dehydrogenase assay at low IC50 values, but agreed well with the Sybr Green plate reader assay and light microscopy. CONCLUSIONS: Although flow cytometric equipment is expensive, the necessary reagents are inexpensive, the procedure is simple and rapid, and the cell volume required is minimal. This should allow field studies using fingerprick sample volumes. PMID- 20003397 TI - The neck-region polymorphism of DC-SIGNR in peri-centenarian from Han Chinese population. AB - BACKGROUND: DC-SIGNR (also called CD209L) has been extensively studied on its role in host genetic predisposition to viral infection. In particular, variable number tandem repeat (VNTR) of the neck-region of DC-SIGNR is highly polymorphic and the polymorphism has been investigated for genetic predisposition to various infectious diseases, though conflicting results had been reported. As infection is a major cause of human death and a mechanism of natural selection, we hypothesized that VNTR polymorphism of DC-SIGNR might have an effect on human life span. METHODS: Here we collected 361 peri-centenarian individuals (age >or=94 for female and age >or=90 for male) and 342 geographically matched controls (age 22-53, mean 35.0 +/- 12.0) from Han Chinese. The VNTR polymorphism of the neck region was determined by PCR and genotype was called by separating the PCR products in agarose gel. RESULTS: A total of 11 genotypes and 5 alleles were found in our population. The genotype distribution, allele frequencies and homozygote proportion did not show a significant difference between peri centenarian and control group. As gender differences in lifespan are ubiquitously observed throughout the animal kingdom, we then stratified the samples by gender. There was more 6/7 genotypes in female peri-centenarian group than that in female control group, at a marginal level of significance (5.56 vs. 1.28%, p = 0.041). The difference was not significant after correction by Bonferroni method. It suggests a possible differential effect of DC-SIGNR VNTR genotypes between sexes. Further studies are warranted to confirm our preliminary findings and investigate the mechanisms of the underlying functions. CONCLUSIONS: Our study indicated that there was absence of association between the neck region polymorphism of DC-SIGNR and longevity in Han Chinese population. But the question of whether the DC-SIGNR could affect longevity in a gender-specific pattern remains open. PMID- 20003399 TI - A typology of practice narratives during the implementation of a preventive, community intervention trial. AB - BACKGROUND: Traditional methods of process evaluation encompass what components were delivered, but rarely uncover how practitioners position themselves and act relative to an intervention being tested. This could be crucial for expanding our understanding of implementation and its contribution to intervention effectiveness. METHODS: We undertook a narrative analysis of in-depth, unstructured field diaries kept by nine community development practitioners for two years. The practitioners were responsible for implementing a multi-component, preventive, community-level intervention for mothers of new babies in eight communities, as part of a cluster randomised community intervention trial. We constructed a narrative typology of approaches to practice, drawing on the phenomenology of Alfred Schutz and Max Weber's Ideal Type theory. RESULTS: Five types of practice emerged, from a highly 'technology-based' type that was faithful to intervention specifications, through to a 'romantic' type that held relationships to be central to daily operations, with intact relationships being the final arbiter of intervention success. The five types also differed in terms of how others involved in the intervention were characterized, the narrative form (e.g., tragedy, satire) and where and how transformative change in communities was best created. This meant that different types traded-off or managed the priorities of the intervention differently, according to the deeply held values of their type. CONCLUSIONS: The data set constructed for this analysis is unique. It revealed that practitioners not only exercise their agency within interventions, they do so systematically, that is, according to a pattern. The typology is the first of its kind and, if verified through replication, may have value for anticipating intervention dynamics and explaining implementation variation in community interventions. PMID- 20003398 TI - The PACE study: a randomised clinical trial of cognitive activity (CA) for older adults with mild cognitive impairment (MCI). AB - BACKGROUND: Research evidence from observational studies suggests that cognitive activity reduces the risk of cognitive impairment in later life as well as the rate of cognitive decline of people with dementia. The Promoting Healthy Ageing with Cognitive Exercise (PACE) study has been designed to determine whether a cognitive activity intervention decreases the rate of cognitive decline amongst older adults with mild cognitive impairment (MCI). METHODS/DESIGN: The study will recruit 160 community-dwelling men and women aged 65 years of age or over with mild cognitive impairment (MCI). Participants will be randomly allocated to two treatment groups: non-specific education and cognitive activity. The intervention will consist of ten 90-minute sessions delivered twice per week over a period of five weeks. The primary outcome measure of the study is the change from baseline in the total score on the Cambridge Cognitive Score (CAMCOG). Secondary outcomes of interest include changes in memory, attention, executive functions, mood and quality of life. Primary endpoints will be collected 12, 52 and 104 weeks after the baseline assessment. DISCUSSION: The proposed project will produce the best available evidence on the merits of increased cognitive activity as a strategy to prevent cognitive decline among older adults with MCI. We anticipate that the results of this study will have implications for the development of evidence based preventive strategies to reduce the rate of cognitive decline amongst older people at risk of dementia. TRIAL REGISTRATION: ACTRN12608000556347. PMID- 20003400 TI - Validation of pooled genotyping on the Affymetrix 500 k and SNP6.0 genotyping platforms using the polynomial-based probe-specific correction. AB - BACKGROUND: The use of pooled DNA on SNP microarrays (SNP-MaP) has been shown to be a cost effective and rapid manner to perform whole-genome association evaluations. While the accuracy of SNP-MaP was extensively evaluated on the early Affymetrix 10 k and 100 k platforms, there have not been as many similarly comprehensive studies on more recent platforms. In the present study, we used the data generated from the full Affymetrix 500 k SNP set together with the polynomial-based probe-specific correction (PPC) to derive allele frequency estimates. These estimates were compared to genotyping results of the same individuals on the same platform, as the basis to evaluate the reliability and accuracy of pooled genotyping on these high-throughput platforms. We subsequently extended this comparison to the new SNP6.0 platform capable of genotyping 1.8 million genetic variants. RESULTS: We showed that pooled genotyping on the 500 k platform performed as well as those previously shown on the relatively lower throughput 10 k and 100 k array sets, with high levels of accuracy (correlation coefficient: 0.988) and low median error (0.036) in allele frequency estimates. Similar results were also obtained from the SNP6.0 array set. A novel pooling strategy of overlapping sub-pools was attempted and comparison of estimated allele frequencies showed this strategy to be as reliable as replicate pools. The importance of an appropriate reference genotyping data set for the application of the PPC algorithm was also evaluated; reference samples with similar ethnic background to the pooled samples were found to improve estimation of allele frequencies. CONCLUSION: We conclude that use of the PPC algorithm to estimate allele frequencies obtained from pooled genotyping on the high throughput 500 k and SNP6.0 platforms is highly accurate and reproducible especially when a suitable reference sample set is used to estimate the beta values for PPC. PMID- 20003401 TI - Universal health care no guarantee of equity: comparison of socioeconomic inequalities in the receipt of coronary procedures in patients with acute myocardial infarction and angina. AB - BACKGROUND: In Australia there is a socioeconomic gradient in morbidity and mortality favouring socioeconomically advantaged people, much of which is accounted for by ischaemic heart disease. This study examines if Australia's universal health care system, with its mixed public/private funding and delivery model, may actually perpetuate this inequity. We do this by quantifying and comparing socioeconomic inequalities in the receipt of coronary procedures in patients with acute myocardial infarction (AMI) and patients with angina. METHODS: Using linked hospital and mortality data, we followed patients admitted to Western Australian hospitals with a first admission for AMI (n = 5539) or angina (n = 7401) in 2001-2003. An outcome event was the receipt, within a year, of a coronary procedure-angiography, angioplasty and/or coronary artery bypass surgery (CABG). Socioeconomic status was assigned to each individual using an area-based measure, the SEIFA Index of Disadvantage. Multivariable proportional hazards regression was used to model the association between socioeconomic status and procedure rates, allowing for censoring and adjustment of multiple covariates. Mediating models examined the effect of private health insurance. RESULTS: In the AMI patient cohort, socioeconomic gradients were not evident except that disadvantaged women were more likely than advantaged women to undergo CABG. In contrast, in the angina patient group there were clear socioeconomic gradients for all procedures, favouring more advantaged patients. Compared with patients in the most disadvantaged quintile of socioeconomic status, patients in the least disadvantaged quintile were 11% (1-21%) more likely to receive angiography, 52% (29-80%) more likely to undergo angioplasty and 30% (3-55%) more likely to undergo CABG. Private health insurance explained some of the socioeconomic variation in rates. CONCLUSIONS: Australia's universal health care system does not guarantee equity in the receipt of high technology health care for patients with ischaemic heart disease. While such a system might ensure equity for patients with AMI, where guidelines for treatment are relatively well established, this is not the case for angina patients, where health care may be less urgent and more discretionary. PMID- 20003403 TI - Associations between successful palliative cancer pathways and community nurse involvement. AB - BACKGROUND: Most terminally ill cancer patients and their relatives wish that the patient dies at home. Community nurses (CNs) are often frontline workers in the patients' homes and CN involvement may be important in attaining successful palliative pathways at home.The aim of the present study was to examine associations between bereaved relatives' evaluation of palliative treatment at home and 1) place of death and 2) CN involvement. METHODS: The study is a population-based, cross-sectional combined register and questionnaire study performed in Aarhus County, Denmark. CN questionnaires were used to obtain data on CNs' efforts, GP-questionnaires were used to obtain data on pathway characteristics and relatives answered questionnaires to evaluate the palliative pathway at home. Questionnaires addressed the palliative pathway of a total of 599 deceased cancer patients. Associations between bereaved relatives' evaluation of palliative pathways at home and place of death and CN involvement were analysed. RESULTS: 'A successful palliative pathway at home' was positively associated with home-death and death at a nursing home compared with death at an institution. No significant associations were identified between the evaluations of the palliative pathway at home and the involvement of CNs. CONCLUSIONS: Our study indicates that dying at home is positively associated with a higher likelihood that the bereaved relative will evaluate the palliative pathway at home as successful. The absence of any significance of involvement of CNs may be ascribed to the variables for involvement chosen in the study. More research is needed on CNs' impact on palliative pathways. PMID- 20003402 TI - Transcriptional profile of Pseudomonas syringae pv. phaseolicola NPS3121 in response to tissue extracts from a susceptible Phaseolus vulgaris L. cultivar. AB - BACKGROUND: Pseudomonas syringae pv. phaseolicola is a Gram-negative plant pathogenic bacterium that causes "halo blight" disease of beans (Phaseolus vulgaris L.). This disease affects both foliage and pods, and is a major problem in temperate areas of the world. Although several bacterial genes have been determined as participants in pathogenesis, the overall process still remains poorly understood, mainly because the identity and function of many of the genes are largely unknown. In this work, a genomic library of P. syringae pv. phaseolicola NPS3121 was constructed and PCR amplification of individual fragments was carried out in order to print a DNA microarray. This microarray was used to identify genes that are differentially expressed when bean leaf extracts, pod extracts or apoplastic fluid were added to the growth medium. RESULTS: Transcription profiles show that 224 genes were differentially expressed, the majority under the effect of bean leaf extract and apoplastic fluid. Some of the induced genes were previously known to be involved in the first stages of the bacterial-plant interaction and virulence. These include genes encoding type III secretion system proteins and genes involved in cell-wall degradation, phaseolotoxin synthesis and aerobic metabolism. On the other hand, most repressed genes were found to be involved in the uptake and metabolism of iron. CONCLUSION: This study furthers the understanding of the mechanisms involved, responses and the metabolic adaptation that occurs during the interaction of P. syringae pv. phaseolicola with a susceptible host plant. PMID- 20003404 TI - Insights into distinct regulatory modes of nucleosome positioning. AB - BACKGROUND: The nucleosome is the fundamental unit of eukaryotic genomes. Experimental evidence suggests that the genomic DNA sequence and a variety of protein factors contribute to nucleosome positioning in vivo. However, how nucleosome positioning is determined locally is still largely unknown. RESULTS: We found that transcription factor binding sites (TFBSs) with particular nucleosomal contexts show a preference to reside on specific chromosomes. We identified four typical gene classes associated with distinct regulatory modes of nucleosome positioning, and further showed that they are distinguished by transcriptional regulation patterns. The first mode involves the cooperativity between chromatin remodeling and stable transcription factor (TF)-DNA binding that is linked to high intrinsic DNA binding affinities, evicting nucleosomes from favorable DNA sequences. The second is the DNA-encoded low nucleosome occupancy that is associated with high gene activity. The third is through chromatin remodeling and histone acetylation, sliding nucleosomes along DNA. This mode is linked to more cryptic sites for TF binding. The last consists of the nucleosome-enriched organization driven by other factors that overrides nucleosome sequence preferences. In addition, we showed that high polymerase II (Pol II) occupancy is associated with high nucleosome occupancy around the transcription start site (TSS). CONCLUSIONS: We identified four different regulatory modes of nucleosome positioning and gave insights into mechanisms that specify promoter nucleosome location. We suggest two distinct modes of recruitment of Pol II, which are selectively employed by different genes. PMID- 20003405 TI - Design of the Balance@Work project: systematic development, evaluation and implementation of an occupational health guideline aimed at the prevention of weight gain among employees. AB - BACKGROUND: Occupational health professionals may play an important role in preventive health promotion activities for employees. However, due to a lack of knowledge and evidence- and practice based methods and strategies, interventions are hardly being implemented by occupational physicians to date. The aim of the Balance@Work project is to develop, evaluate, and implement an occupational health guideline aimed at the prevention of weight gain among employees. METHODS: Following the guideline development protocol of the Netherlands Society of Occupational Medicine and the Intervention Mapping protocol, the guideline was developed based on literature, interviews with relevant stakeholders, and consensus among an expert group. The guideline consists of an individual and an environmental component. The individual component includes recommendations for occupational physicians on how to promote physical activity and healthy dietary behavior based on principles of motivational interviewing. The environmental component contains an obesogenic environment assessment tool. The guideline is evaluated in a randomised controlled trial among 20 occupational physicians. Occupational physicians in the intervention group apply the guideline to eligible workers during 6 months. Occupational physicians in the control group provide care as usual. Measurements take place at baseline and 6, 12, and 18 months thereafter. Primary outcome measures include waist circumference, daily physical activity and dietary behavior. Secondary outcome measures include sedentary behavior, determinants of behavior change, body weight and body mass index, cardiovascular disease risk profile, and quality of life. Additionally, productivity, absenteeism, and cost-effectiveness are assessed. DISCUSSION: Improving workers' daily physical activity and dietary behavior may prevent weight gain and subsequently improve workers' health, increase productivity, and reduce absenteeism. After an effect- and process evaluation the guideline will be adjusted and, after authorisation, published. Together with several implementation aids, the published guideline will be disseminated broadly by the Netherlands Society of Occupational Medicine. TRIAL REGISTRATION: ISRCTN73545254/NTR1190. PMID- 20003406 TI - Gender differences in presentation and diagnosis of chest pain in primary care. AB - BACKGROUND: Chest pain is a common complaint and reason for consultation in primary care. Research related to gender differences in regard to Coronary Heart Disease (CHD) has been mainly conducted in hospital but not in primary care settings. We aimed to analyse gender differences in aetiology and clinical characteristics of chest pain and to provide gender related symptoms and signs associated with CHD. METHODS: We included 1212 consecutive patients with chest pain aged 35 years and older attending 74 general practitioners (GPs). GPs recorded symptoms and findings of each patient and provided follow up information. An independent interdisciplinary reference panel reviewed clinical data of every patient and decided about the aetiology of chest pain at the time of patient recruitment. Multivariable regression analysis was performed to identify clinical predictors that help to rule in or out CHD in women and men. RESULTS: Women showed more psychogenic disorders (women 11,2%, men 7.3%, p = 0.02), men suffered more from CHD (women 13.0%, men 17.2%, p = 0.04), trauma (women 1.8%, men 5.1%, p < 0.001) and pneumonia/pleurisy (women 1.3%, men 3.0%, p = 0.04) Men showed significantly more often chest pain localised on the right side of the chest (women 9.1%, men 25.0%, p = 0.01). For both genders known clinical vascular disease, pain worse with exercise and age were associated positively with CHD. In women pain duration above one hour was associated positively with CHD, while shorter pain durations showed an association with CHD in men. In women negative associations were found for stinging pain and in men for pain depending on inspiration and localised muscle tension. CONCLUSIONS: We found gender differences in regard to aetiology, selected clinical characteristics and association of symptoms and signs with CHD in patients presenting with chest pain in a primary care setting. Further research is necessary to elucidate whether these differences would support recommendations for different diagnostic approaches for CHD according to a patient's gender. PMID- 20003407 TI - The polycyclic aromatic hydrocarbon concentrations in soils in the Region of Valasske Mezirici, the Czech Republic. AB - The polycyclic aromatic hydrocarbon (PAH) contamination of urban, agricultural and forest soil samples was investigated from samples obtained in the surroundings of Valasske Mezirici. Valasske Mezirici is a town located in the north-east mountainous part of the Czech Republic, where a coal tar refinery is situated. 16 PAHs listed in the US EPA were investigated. Organic oxidizable carbon was also observed in the forest soils. The PAH concentrations ranged from 0.86-10.84 (with one anomalous value of 35.14) and 7.66-79.39 mg/kg dm in the urban/agricultural and forest soils, respectively. While the PAH levels in the urban/agricultural soils are within the range typically found in industrialized areas, the forest soils showed elevated PAH concentrations compared to other forest soils in Western and Northern Europe. The PAH concentrations and their molecular distribution ratios were studied as functions of the sample location and the meteorological history. The soils from localities at higher altitudes above sea level have the highest PAH concentrations, and the PAH concentrations decrease with increasing distance from the town. PMID- 20003408 TI - A systems analysis of the chemosensitivity of breast cancer cells to the polyamine analogue PG-11047. AB - BACKGROUND: Polyamines regulate important cellular functions and polyamine dysregulation frequently occurs in cancer. The objective of this study was to use a systems approach to study the relative effects of PG-11047, a polyamine analogue, across breast cancer cells derived from different patients and to identify genetic markers associated with differential cytotoxicity. METHODS: A panel of 48 breast cell lines that mirror many transcriptional and genomic features present in primary human breast tumours were used to study the antiproliferative activity of PG-11047. Sensitive cell lines were further examined for cell cycle distribution and apoptotic response. Cell line responses, quantified by the GI50 (dose required for 50% relative growth inhibition) were correlated with the omic profiles of the cell lines to identify markers that predict response and cellular functions associated with drug sensitivity. RESULTS: The concentrations of PG-11047 needed to inhibit growth of members of the panel of breast cell lines varied over a wide range, with basal-like cell lines being inhibited at lower concentrations than the luminal cell lines. Sensitive cell lines showed a significant decrease in S phase fraction at doses that produced little apoptosis. Correlation of the GI50 values with the omic profiles of the cell lines identified genomic, transcriptional and proteomic variables associated with response. CONCLUSIONS: A 13-gene transcriptional marker set was developed as a predictor of response to PG-11047 that warrants clinical evaluation. Analyses of the pathways, networks and genes associated with response to PG-11047 suggest that response may be influenced by interferon signalling and differential inhibition of aspects of motility and epithelial to mesenchymal transition. PMID- 20003409 TI - Integrating complex genomic datasets and tumour cell sensitivity profiles to address a 'simple' question: which patients should get this drug? AB - It is becoming increasingly apparent that cancer drug therapies can only reach their full potential through appropriate patient selection. Matching drugs and cancer patients has proven to be a complex challenge, due in large part to the substantial molecular heterogeneity inherent to human cancers. This is not only a major hurdle to the improvement of the use of current treatments but also for the development of novel therapies and the ability to steer them to the relevant clinical indications. In this commentary we discuss recent studies from Kuo et al., published this month in BMC Medicine, in which they used a panel of cancer cell lines as a model for capturing patient heterogeneity at the genomic and proteomic level in order to identify potential biomarkers for predicting the clinical activity of a novel candidate chemotherapeutic across a patient population. The findings highlight the ability of a 'systems approach' to develop a better understanding of the properties of novel candidate therapeutics and to guide clinical testing and application.See the associated research paper by Kuo et al: http://www.biomedcentral.com/1741-7015/7/77. PMID- 20003410 TI - Characterization of the histone H2A.Z-1 and H2A.Z-2 isoforms in vertebrates. AB - BACKGROUND: Within chromatin, the histone variant H2A.Z plays a role in many diverse nuclear processes including transcription, preventing the spread of heterochromatin and epigenetic transcriptional memory. The molecular mechanisms of how H2A.Z mediates its effects are not entirely understood. However, it is now known that H2A.Z has two protein isoforms in vertebrates, H2A.Z-1 and H2A.Z-2, which are encoded by separate genes and differ by 3 amino acid residues. RESULTS: We report that H2A.Z-1 and H2A.Z-2 are expressed across a wide range of human tissues, they are both acetylated at lysine residues within the N-terminal region and they exhibit similar, but nonidentical, distributions within chromatin. Our results suggest that H2A.Z-2 preferentially associates with H3 trimethylated at lysine 4 compared to H2A.Z-1. The phylogenetic analysis of the promoter regions of H2A.Z-1 and H2A.Z-2 indicate that they have evolved separately during vertebrate evolution. CONCLUSIONS: Our biochemical, gene expression, and phylogenetic data suggest that the H2A.Z-1 and H2A.Z-2 variants function similarly yet they may have acquired a degree of functional independence. PMID- 20003411 TI - Factors associated with maternal mortality in Sub-Saharan Africa: an ecological study. AB - BACKGROUND: Maternal health is one of the major worldwide health challenges. Currently, the unacceptably high levels of maternal mortality are a common subject in global health and development discussions. Although some countries have made remarkable progress, half of the maternal deaths in the world still take place in Sub-Saharan Africa where little or no progress has been made. There is no single simple, straightforward intervention that will significantly decrease maternal mortality alone; however, there is a consensus on the importance of a strong health system, skilled delivery attendants, and women's rights for maternal health. Our objective was to describe and determine different factors associated with the maternal mortality ratio in Sub-Saharan countries. METHODS: An ecological multi-group study compared variables between many countries in Sub-Saharan Africa using data collected between 1997 and 2006. The dependent variable was the maternal mortality ratio, and Health care system related, educational and economic indicators were the independent variables. Information sources included the WHO, World Bank, UNICEF and UNDP. RESULTS: Maternal mortality ratio values in Sub-Saharan Africa were demonstrated to be high and vary enormously among countries. A relationship between the maternal mortality ratio and some educational, sanitary and economic factors was observed. There was an inverse and significant correlation of the maternal mortality ratio with prenatal care coverage, births assisted by skilled health personnel, access to an improved water source, adult literacy rate, primary female enrolment rate, education index, the Gross National Income per capita and the per-capita government expenditure on health. CONCLUSIONS: Education and an effective and efficient health system, especially during pregnancy and delivery, are strongly related to maternal death. Also, macro-economic factors are related and could be influencing the others. PMID- 20003412 TI - Prediction of seizure control in non-ketotic hyperglycemic induced seizures. AB - BACKGROUND: To study the factors predictive for seizure control in non-ketotic hyperglycemic induced seizures (NKHS). METHODS: We studied 21 patients who were clinically diagnosed as NKHS at Khon Kaen University hospital, Thailand. Multiple linear regression analysis was used to identify the factors predictive for seizure control. RESULTS: Most patients had no previous history of diabetes and presented with repetitive partial seizures. The mean number of seizure attacks was 45 times prior to admission. The average duration to terminate seizure was 36 hours and significantly predicted by frequency of seizures (estimate 0.9, p value 0.013). CONCLUSION: Frequency of seizures is the only predictive factor for the success of seizure control in NKHS. PMID- 20003413 TI - Nuclear factor I-A represses expression of the cell adhesion molecule L1. AB - BACKGROUND: The neural cell adhesion molecule L1 plays a crucial role in development and plasticity of the nervous system. Neural cells thus require precise control of L1 expression. RESULTS: We identified a full binding site for nuclear factor I (NFI) transcription factors in the regulatory region of the mouse L1 gene. Electrophoretic mobility shift assay (EMSA) showed binding of nuclear factor I-A (NFI-A) to this site. Moreover, for a brain-specific isoform of NFI-A (NFI-A bs), we confirmed the interaction in vivo using chromatin immunoprecipitation (ChIP). Reporter gene assays showed that in neuroblastoma cells, overexpression of NFI-A bs repressed L1 expression threefold. CONCLUSION: Our findings suggest that NFI-A, in particular its brain-specific isoform, represses L1 gene expression, and might act as a second silencer of L1 in addition to the neural restrictive silencer factor (NRSF). PMID- 20003414 TI - Effects of normalization on quantitative traits in association test. AB - BACKGROUND: Quantitative trait loci analysis assumes that the trait is normally distributed. In reality, this is often not observed and one strategy is to transform the trait. However, it is not clear how much normality is required and which transformation works best in association studies. RESULTS: We performed simulations on four types of common quantitative traits to evaluate the effects of normalization using the logarithm, Box-Cox, and rank-based transformations. The impact of sample size and genetic effects on normalization is also investigated. Our results show that rank-based transformation gives generally the best and consistent performance in identifying the causal polymorphism and ranking it highly in association tests, with a slight increase in false positive rate. CONCLUSION: For small sample size or genetic effects, the improvement in sensitivity for rank transformation outweighs the slight increase in false positive rate. However, for large sample size and genetic effects, normalization may not be necessary since the increase in sensitivity is relatively modest. PMID- 20003415 TI - Global end-diastolic volume acquired by transpulmonary thermodilution depends on age and gender in awake and spontaneously breathing patients. AB - INTRODUCTION: Volumetric parameters acquired by transpulmonary thermodilution had been repeatedly proven superior to filling pressures for estimation of cardiac preload. Up to now, the proposed normal ranges were never studied in detail. We investigated the relationship of the global end-diastolic volume (GEDV) acquired by transpulmonary thermodilution with age and gender in awake and spontaneously breathing patients. METHODS: Patients requiring brain tumor surgery were equipped prospectively with a transpulmonary thermodilution device. On postoperative day one, thermodilution measurements were performed in 101 patients ready for discharge from the ICU. All subjects were awake, spontaneously breathing, hemodynamically stable and free of catecholamines. RESULTS: Main finding was a dependence of GEDV on age and gender, height and weight of the patient. Age was a highly significant non-linear coefficient for GEDV with large inter-individual variance (p < 0.001). On average, GEDV was 131.1 ml higher in males (p = 0.027). Each cm body height accounted for 13.0 ml additional GEDV (p < 0.001). GEDV increased by 2.90 ml per kg actual body weight (p = 0.043). Each cofactor, including height and weight, remained significant after indexing GEDV to body surface area using predicted body weight. CONCLUSIONS: The volumetric parameter GEDV shows a large inter-individual variance and is dependent on age and gender. These dependencies persist after indexing GEDV to body surface area calculated with predicted body weight. Targeting resuscitation using fixed ranges of preload volumes acquired by transpulmonary thermodilution without concern to an individual patient's age and gender seems not to be appropriate. PMID- 20003416 TI - STK39 polymorphisms and blood pressure: an association study in British Caucasians and assessment of cis-acting influences on gene expression. AB - BACKGROUND: Blood pressure (BP) has significant heritability, but the genes responsible remain largely unknown. Single nucleotide polymorphisms (SNPs) at the STK39 locus were recently associated with hypertension by genome-wide association in an Amish population; in vitro data from transient transfection experiments using reporter constructs suggested that altered STK39 expression might mediate the effect. However, other large studies have not implicated STK39 in hypertension. We determined whether reported SNPs influenced STK39 expression in vivo, or were associated with BP in a large British Caucasian cohort. METHODS: 1372 members of 247 Caucasian families ascertained through a hypertensive proband were genotyped for reported risk variants in STK39 (rs6749447, rs3754777, rs35929607) using Sequenom technology. MERLIN software was used for family-based association testing. Cis-acting influences on expression were assessed in vivo using allelic expression ratios in cDNA from peripheral blood cells in 35 South African individuals heterozygous for a transcribed SNP in STK39 (rs1061471) and quantified by mass spectrometry (Sequenom). RESULTS: No significant association was seen between the SNPs tested and systolic or diastolic BP in clinic or ambulatory measurements (all p > 0.05). The tested SNPs were all associated with allelic expression differences in peripheral blood cells (p < 0.05), with the most significant association for the intronic SNP rs6749447 (P = 9.9 x 10-4). In individuals who were heterozygous for this SNP, on average the G allele showed 13% overexpression compared to the T allele. CONCLUSIONS: STK39 expression is modified by polymorphisms acting in cis and the typed SNPs are associated with allelic expression of this gene, but there is no evidence for an association with BP in a British Caucasian cohort. PMID- 20003417 TI - Assessing functional mitral regurgitation with exercise echocardiography: rationale and clinical applications. AB - Secondary or functional mitral regurgitation (FMR) represents an increasing feature of mitral valve disease characterized by abnormal function of anatomically normal leaflets in the context of the impaired function of remodelled left ventricles. The anatomic and pathophysiological basis of FMR are briefly analyzed; in addition, the role of exercise echocardiography for the assessment of FMR is discussed in view of its relevance to clinical practice. PMID- 20003418 TI - Combination effect of antithrombin and recombinant human soluble thrombomodulin in a lipopolysaccharide induced rat sepsis model. AB - INTRODUCTION: Recombinant human soluble thrombomodulin (rhsTM) is newly developed for the treatment of DIC. The purpose of this study was to evaluate the efficacy of the concomitant administration of rhsTM and antithrombin (AT). METHODS: In the first series, rats were treated with either 62.5, 125, 250 or 500 IU/kg (n = 6, each) of AT or 0.125, 0.25, 0.5 or 1.0 mg/kg (n = 6, each) of rhsTM followed by lipopolysaccharide (LPS) injection. 8 h later, the fibrinogen level was examined. In the second series, TM group was pretreated with 0.25 mg/kg of rhsTM, AT group was pretreated with 125 IU/kg of AT, AT/TM group was pretreated with both AT and rhsTM, and control group was pretreated with saline (n = 7, each). The platelet count, fibrinogen, ALT, LDH and high-mobility group box 1 (HMGB1) levels were measured. In addition, histologic changes in liver were examined. In the third series, survival was calculated up to 24 h. RESULTS: Both AT and rhsTM produced a linear dose-response with regard to the fibrinogen level, with 125 IU/kg of AT and 0.25 mg/kg of rhsTM producing equivalent effects. The combined administration of AT and rhsTM significantly reduced the decrease in the platelet count and the fibrinogen level (P < 0.05, 0.01, respectively). The elevations in ALT and LDH were significantly suppressed in all treatment groups. The HMGB1 level and the histologic changes tended to indicate damage reduction. Survival was significantly better only in AT/TM group (P < 0.01). CONCLUSIONS: The coadministration of AT and rhsTM might be effective for the treatment of severe sepsis. PMID- 20003419 TI - The mitochondrial genome of the 'twisted-wing parasite' Mengenilla australiensis (Insecta, Strepsiptera): a comparative study. AB - BACKGROUND: Strepsiptera are an unusual group of sexually dimorphic, entomophagous parasitoids whose evolutionary origins remain elusive. The lineage leading to Mengenilla australiensis (Family Mengenillidae) is the sister group to all remaining extant strepsipterans. It is unique in that members of this family have retained a less derived condition, where females are free-living from pupation onwards, and are structurally much less simplified. We sequenced almost the entire mitochondrial genome of M. australiensis as an important comparative data point to the already available genome of its distant relative Xenos vesparum (Family Xenidae). This study represents the first in-depth comparative mitochondrial genomic analysis of Strepsiptera. RESULTS: The partial genome of M. australiensis is presented as a 13421 bp fragment, across which all 13 protein coding genes (PCGs), 2 ribosomal RNA (rRNA) genes and 18 transfer RNA (tRNA) sequences are identified. Two tRNA translocations disrupt an otherwise ancestral insect mitochondrial genome order. A+T content is measured at 84.3%, C-content is also very skewed. Compared with M. australiensis, codon bias in X. vesparum is more balanced. Interestingly, the size of the protein coding genome is truncated in both strepsipterans, especially in X. vesparum which, uniquely, has 4.3% fewer amino acids than the average holometabolan complement. A revised assessment of mitochondrial rRNA secondary structure based on comparative structural considerations is presented for M. australiensis and X. vesparum. CONCLUSIONS: The mitochondrial genome of X. vesparum has undergone a series of alterations which are probably related to an extremely derived lifestyle. Although M. australiensis shares some of these attributes; it has retained greater signal from the hypothetical most recent common ancestor (MRCA) of Strepsiptera, inviting the possibility that a shift in the mitochondrial selective environment might be related to the specialization accompanying the evolution of a small, morphologically simplified completely host-dependent lifestyle. These results provide useful insights into the nature of the evolutionary transitions that accompanied the emergence of Strepsiptera, but we emphasize the need for adequate sampling across the order in future investigations concerning the extraordinary developmental and evolutionary origins of this group. PMID- 20003420 TI - Transpositionally active episomal hAT elements. AB - BACKGROUND: hAT elements and V(D)J recombination may have evolved from a common ancestral transposable element system. Extrachromosomal, circular forms of transposable elements (referred to here as episomal forms) have been reported yet their biological significance remains unknown. V(D)J signal joints, which resemble episomal transposable elements, have been considered non-recombinogenic products of V(D)J recombination and a safe way to dispose of excised chromosomal sequences. V(D)J signal joints can, however, participate in recombination reactions and the purpose of this study was to determine if hobo and Hermes episomal elements are also recombinogenic. RESULTS: Up to 50% of hobo/Hermes episomes contained two intact, inverted-terminal repeats and 86% of these contained from 1-1000 bp of intercalary DNA. Episomal hobo/Hermes elements were recovered from Musca domestica (a natural host of Hermes), Drosophila melanogaster (a natural host of hobo) and transgenic Drosophila melanogaster and Aedes aegypti (with autonomous Hermes elements). Episomal Hermes elements were recovered from unfertilized eggs of M. domestica and D. melanogaster demonstrating their potential for extrachromosomal, maternal transmission. Reintegration of episomal Hermes elements was observed in vitro and in vivo and the presence of Hermes episomes resulted in lower rates of canonical Hermes transposition in vivo. CONCLUSION: Episomal hobo/Hermes elements are common products of element excision and can be maternally transmitted. Episomal forms of Hermes are capable of integration and also of influencing the transposition of canonical elements suggesting biological roles for these extrachromosomal elements in element transmission and regulation. PMID- 20003422 TI - Towards equitable access to medicines for the rural poor: analyses of insurance claims reveal rural pharmacy initiative triggers price competition in Kyrgyzstan. AB - BACKGROUND: A rural pharmacy initiative (RPI) designed to increase access to medicines in rural Kyrgyzstan created a network of 12 pharmacies using a revolving drug fund mechanism in 12 villages where no pharmacies previously existed. The objective of this study was to determine if the establishment of the RPI resulted in the unforeseen benefit of triggering medicine price competition in pre-existing (non-RPI) private pharmacies located in the region. METHODS: We conducted descriptive and multivariate analyses on medicine insurance claims data from Kyrgyzstan's Mandatory Health Insurance Fund for the Jumgal District of Naryn Province from October 2003 to December 2007. We compared average quarterly medicine prices in competitor pharmacies before and after the introduction of the rural pharmacy initiative in October 2004 to determine the RPI impact on price competition. RESULTS: Descriptive analyses suggest competitors reacted to RPI prices for 21 of 30 (70%) medicines. Competitor medicine prices from the quarter before RPI introduction to the end of the study period decreased for 17 of 30 (57%) medicines, increased for 4 of 30 (13%) medicines, and remained unchanged for 9 of 30 (30%) medicines. Among the 9 competitor medicines with unchanged prices, five initially decreased in price but later reverted back to baseline prices. Multivariate analyses on 19 medicines that met sample size criteria confirm these findings. Fourteen of these 19 (74%) competitor medicines changed significantly in price from the quarter before RPI introduction to the quarter after RPI introduction, with 9 of 19 (47%) decreasing in price and 5 of 19 (26%) increasing in price. CONCLUSIONS: The RPI served as a market driver, spurring competition in medicine prices in competitor pharmacies, even when they were located in different villages. Initiatives designed to increase equitable access to medicines in rural regions of developing and transitional countries should consider the potential to leverage medicine price competition as a means of achieving their goal. Evaluations of interventions to increase rural access to medicines should include impact assessment on both formal and informal pharmaceutical markets. PMID- 20003421 TI - Recombinant protein expression by targeting pre-selected chromosomal loci. AB - BACKGROUND: Recombinant protein expression in mammalian cells is mostly achieved by stable integration of transgenes into the chromosomal DNA of established cell lines. The chromosomal surroundings have strong influences on the expression of transgenes. The exploitation of defined loci by targeting expression constructs with different regulatory elements is an approach to design high level expression systems. Further, this allows to evaluate the impact of chromosomal surroundings on distinct vector constructs. RESULTS: We explored antibody expression upon targeting diverse expression constructs into previously tagged loci in CHO-K1 and HEK293 cells that exhibit high reporter gene expression. These loci were selected by random transfer of reporter cassettes and subsequent screening. Both, retroviral infection and plasmid transfection with eGFP or antibody expression cassettes were employed for tagging. The tagged cell clones were screened for expression and single copy integration. Cell clones producing > 20 pg/cell in 24 hours could be identified. Selected integration sites that had been flanked with heterologous recombinase target sites (FRTs) were targeted by Flp recombinase mediated cassette exchange (RMCE). The results give proof of principle for consistent protein expression upon RMCE. Upon targeting antibody expression cassettes 90-100% of all resulting cell clones showed correct integration. Antibody production was found to be highly consistent within the individual cell clones as expected from their isogenic nature. However, the nature and orientation of expression control elements revealed to be critical. The impact of different promoters was examined with the tag-and-targeting approach. For each of the chosen promoters high expression sites were identified. However, each site supported the chosen promoters to a different extent, indicating that the strength of a particular promoter is dominantly defined by its chromosomal context. CONCLUSION: RMCE provides a powerful method to specifically design vectors for optimized gene expression with high accuracy. Upon considering the specific requirements of chromosomal sites this method provides a unique tool to exploit such sites for predictable expression of biotechnologically relevant proteins such as antibodies. PMID- 20003423 TI - Mechanism of primitive duct formation in the pancreas and submandibular glands: a role for SDF-1. AB - BACKGROUND: The exocrine pancreas is composed of a branched network of ducts connected to acini. They are lined by a monolayered epithelium that derives from the endoderm and is surrounded by mesoderm-derived mesenchyme. The morphogenic mechanisms by which the ductal network is established as well as the signaling pathways involved in this process are poorly understood. RESULTS: By morphological analyzis of wild-type and mutant mouse embryos and using cultured embryonic explants we investigated how epithelial morphogenesis takes place and is regulated by chemokine signaling. Pancreas ontogenesis displayed a sequence of two opposite epithelial transitions. During the first transition, the monolayered and polarized endodermal cells give rise to tissue buds composed of a mass of non polarized epithelial cells. During the second transition the buds reorganize into branched and polarized epithelial monolayers that further differentiate into tubulo-acinar glands. We found that the second epithelial transition is controlled by the chemokine Stromal cell-Derived Factor (SDF)-1. The latter is expressed by the mesenchyme, whereas its receptor CXCR4 is expressed by the epithelium. Reorganization of cultured pancreatic buds into monolayered epithelia was blocked in the presence of AMD3100, a SDF-1 antagonist. Analyzis of sdf1 and cxcr4 knockout embryos at the stage of the second epithelial transition revealed transient defective morphogenesis of the ventral and dorsal pancreas. Reorganization of a globular mass of epithelial cells in polarized monolayers is also observed during submandibular glands development. We found that SDF-1 and CXCR4 are expressed in this organ and that AMD3100 treatment of submandibular gland explants blocks its branching morphogenesis. CONCLUSION: In conclusion, our data show that the primitive pancreatic ductal network, which is lined by a monolayered and polarized epithelium, forms by remodeling of a globular mass of non polarized epithelial cells. Our data also suggest that SDF-1 controls the branching morphogenesis of several exocrine tissues. PMID- 20003424 TI - A high-density collection of EMS-induced mutations for TILLING in Landsberg erecta genetic background of Arabidopsis. AB - BACKGROUND: Arabidopsis thaliana is the main model species for plant molecular genetics studies and world-wide efforts are devoted to identify the function of all its genes. To this end, reverse genetics by TILLING (Targeting Induced Local Lesions IN Genomes) in a permanent collection of chemically induced mutants is providing a unique resource in Columbia genetic background. In this work, we aim to extend TILLING resources available in A. thaliana by developing a new population of ethyl methanesulphonate (EMS) induced mutants in the second commonest reference strain. In addition, we pursue to saturate the number of EMS induced mutations that can be tolerated by viable and fertile plants. RESULTS: By mutagenizing with different EMS concentrations we have developed a permanent collection of 3712 M2/M3 independent mutant lines in the reference strain Landsberg erecta (Ler) of A. thaliana. This population has been named as the Arabidopsis TILLer collection. The frequency of mutations per line was maximized by using M1 plants with low but sufficient seed fertility. Application of TILLING to search for mutants in 14 genes identified 21 to 46 mutations per gene, which correspond to a total of 450 mutations. Missense mutations were found for all genes while truncations were selected for all except one. We estimated that, on average, these lines carry one mutation every 89 kb, Ler population providing a total of more than five million induced mutations. It is estimated that TILLer collection shows a two to three fold higher EMS mutation density per individual than previously reported A. thaliana population. CONCLUSIONS: Analysis of TILLer collection demonstrates its usefulness for large scale TILLING reverse genetics in another reference genetic background of A. thaliana. Comparisons with TILLING populations in other organisms indicate that this new A. thaliana collection carries the highest chemically induced mutation density per individual known in diploid species. PMID- 20003425 TI - The other side of comparative genomics: genes with no orthologs between the cow and other mammalian species. AB - BACKGROUND: With the rapid growth in the availability of genome sequence data, the automated identification of orthologous genes between species (orthologs) is of fundamental importance to facilitate functional annotation and studies on comparative and evolutionary genomics. Genes with no apparent orthologs between the bovine and human genome may be responsible for major differences between the species, however, such genes are often neglected in functional genomics studies. RESULTS: A BLAST-based method was exploited to explore the current annotation and orthology predictions in Ensembl. Genes with no orthologs between the two genomes were classified into groups based on alignments, ontology, manual curation and publicly available information. Starting from a high quality and specific set of orthology predictions, as provided by Ensembl, hidden relationship between genes and genomes of different mammalian species were unveiled using a highly sensitive approach, based on sequence similarity and genomic comparison. CONCLUSIONS: The analysis identified 3,801 bovine genes with no orthologs in human and 1010 human genes with no orthologs in cow, among which 411 and 43 genes, respectively, had no match at all in the other species. Most of the apparently non-orthologous genes may potentially have orthologs which were missed in the annotation process, despite having a high percentage of identity, because of differences in gene length and structure. The comparative analysis reported here identified gene variants, new genes and species-specific features and gave an overview of the other side of orthology which may help to improve the annotation of the bovine genome and the knowledge of structural differences between species. PMID- 20003426 TI - DescFold: a web server for protein fold recognition. AB - BACKGROUND: Machine learning-based methods have been proven to be powerful in developing new fold recognition tools. In our previous work [Zhang, Kochhar and Grigorov (2005) Protein Science, 14: 431-444], a machine learning-based method called DescFold was established by using Support Vector Machines (SVMs) to combine the following four descriptors: a profile-sequence-alignment-based descriptor using Psi-blast e-values and bit scores, a sequence-profile-alignment based descriptor using Rps-blast e-values and bit scores, a descriptor based on secondary structure element alignment (SSEA), and a descriptor based on the occurrence of PROSITE functional motifs. In this work, we focus on the improvement of DescFold by incorporating more powerful descriptors and setting up a user-friendly web server. RESULTS: In seeking more powerful descriptors, the profile-profile alignment score generated from the COMPASS algorithm was first considered as a new descriptor (i.e., PPA). When considering a profile-profile alignment between two proteins in the context of fold recognition, one protein is regarded as a template (i.e., its 3D structure is known). Instead of a sequence profile derived from a Psi-blast search, a structure-seeded profile for the template protein was generated by searching its structural neighbors with the assistance of the TM-align structural alignment algorithm. Moreover, the COMPASS algorithm was used again to derive a profile-structural-profile-alignment-based descriptor (i.e., PSPA). We trained and tested the new DescFold in a total of 1,835 highly diverse proteins extracted from the SCOP 1.73 version. When the PPA and PSPA descriptors were introduced, the new DescFold boosts the performance of fold recognition substantially. Using the SCOP_1.73_40% dataset as the fold library, the DescFold web server based on the trained SVM models was further constructed. To provide a large-scale test for the new DescFold, a stringent test set of 1,866 proteins were selected from the SCOP 1.75 version. At a less than 5% false positive rate control, the new DescFold is able to correctly recognize structural homologs at the fold level for nearly 46% test proteins. Additionally, we also benchmarked the DescFold method against several well-established fold recognition algorithms through the LiveBench targets and Lindahl dataset. CONCLUSIONS: The new DescFold method was intensively benchmarked to have very competitive performance compared with some well-established fold recognition methods, suggesting that it can serve as a useful tool to assist in template based protein structure prediction. The DescFold server is freely accessible at http://202.112.170.199/DescFold/index.html. PMID- 20003427 TI - No difference in between-country variability in use of newly approved orphan and non- orphan medicinal products--a pilot study. AB - BACKGROUND: Regulators and payers have to strike a balance between the needs of the patient and the optimal allocation of resources. Drugs indicated for rare diseases (orphan medicines) are a special group in this context because of their often high per unit costs. Our objective in this pilot study was to determine, for drugs used in an outpatient setting, how utilisation of centrally authorised drugs varies between countries across a selection of EU member states. METHODS: We randomly selected five orphan medicines and nine other drugs that were centrally authorised in the European Union between January 2000 and November 2006. We compared utilisation of these drugs in six European Union member states: Austria, Denmark, Finland, Portugal, The Netherlands, and Sweden. Utilisation data were expressed as Defined Daily Doses per 1000 persons per year. Variability in use across countries was determined by calculating the relative standard deviation for the utilisation rates of individual drugs across countries. RESULTS: No association between orphan medicine status and variability in use across countries was found (P = 0.52). Drugs with an orphan medicine status were more expensive and had a higher innovation score than drugs without an orphan medicine status. CONCLUSIONS: The results show that the variability in use of orphan medicines in the different health care systems of the European Union appears to be comparable to the other newly authorised drugs that were included in the analysis. This means that, although strong heterogeneity in access may exist, this heterogeneity is not specific for drugs with an orphan status. PMID- 20003428 TI - Estimates of linkage disequilibrium and effective population size in rainbow trout. AB - BACKGROUND: The use of molecular genetic technologies for broodstock management and selective breeding of aquaculture species is becoming increasingly more common with the continued development of genome tools and reagents. Several laboratories have produced genetic maps for rainbow trout to aid in the identification of loci affecting phenotypes of interest. These maps have resulted in the identification of many quantitative/qualitative trait loci affecting phenotypic variation in traits associated with albinism, disease resistance, temperature tolerance, sex determination, embryonic development rate, spawning date, condition factor and growth. Unfortunately, the elucidation of the precise allelic variation and/or genes underlying phenotypic diversity has yet to be achieved in this species having low marker densities and lacking a whole genome reference sequence. Experimental designs which integrate segregation analyses with linkage disequilibrium (LD) approaches facilitate the discovery of genes affecting important traits. To date the extent of LD has been characterized for humans and several agriculturally important livestock species but not for rainbow trout. RESULTS: We observed that the level of LD between syntenic loci decayed rapidly at distances greater than 2 cM which is similar to observations of LD in other agriculturally important species including cattle, sheep, pigs and chickens. However, in some cases significant LD was also observed up to 50 cM. Our estimate of effective population size based on genome wide estimates of LD for the NCCCWA broodstock population was 145, indicating that this population will respond well to high selection intensity. However, the range of effective population size based on individual chromosomes was 75.51 - 203.35, possibly indicating that suites of genes on each chromosome are disproportionately under selection pressures. CONCLUSIONS: Our results indicate that large numbers of markers, more than are currently available for this species, will be required to enable the use of genome-wide integrated mapping approaches aimed at identifying genes of interest in rainbow trout. PMID- 20003429 TI - Graphle: Interactive exploration of large, dense graphs. AB - BACKGROUND: A wide variety of biological data can be modeled as network structures, including experimental results (e.g. protein-protein interactions), computational predictions (e.g. functional interaction networks), or curated structures (e.g. the Gene Ontology). While several tools exist for visualizing large graphs at a global level or small graphs in detail, previous systems have generally not allowed interactive analysis of dense networks containing thousands of vertices at a level of detail useful for biologists. Investigators often wish to explore specific portions of such networks from a detailed, gene-specific perspective, and balancing this requirement with the networks' large size, complex structure, and rich metadata is a substantial computational challenge. RESULTS: Graphle is an online interface to large collections of arbitrary undirected, weighted graphs, each possibly containing tens of thousands of vertices (e.g. genes) and hundreds of millions of edges (e.g. interactions). These are stored on a centralized server and accessed efficiently through an interactive Java applet. The Graphle applet allows a user to examine specific portions of a graph, retrieving the relevant neighborhood around a set of query vertices (genes). This neighborhood can then be refined and modified interactively, and the results can be saved either as publication-quality images or as raw data for further analysis. The Graphle web site currently includes several hundred biological networks representing predicted functional relationships from three heterogeneous data integration systems: S. cerevisiae data from bioPIXIE, E. coli data using MEFIT, and H. sapiens data from HEFalMp. CONCLUSIONS: Graphle serves as a search and visualization engine for biological networks, which can be managed locally (simplifying collaborative data sharing) and investigated remotely. The Graphle framework is freely downloadable and easily installed on new servers, allowing any lab to quickly set up a Graphle site from which their own biological network data can be shared online. PMID- 20003430 TI - 'Who's who' in two different flower types of Calluna vulgaris (Ericaceae): morphological and molecular analyses of flower organ identity. AB - BACKGROUND: The ornamental crop Calluna vulgaris is of increasing importance to the horticultural industry in the northern hemisphere due to a flower organ mutation: the flowers of the 'bud-flowering' phenotype remain closed i.e. as buds throughout the total flowering period and thereby maintain more colorful flowers for a longer period of time than the wild-type. This feature is accompanied and presumably caused by the complete lack of stamens. Descriptions of this botanical particularity are inconsistent and partially conflicting. In order to clarify basic questions of flower organ identity in general and stamen loss in detail, a study of the wild-type and the 'bud-flowering' flower type of C. vulgaris was initiated. RESULTS: Flowers were examined by macro- and microscopic techniques. Organ development was investigated comparatively in both the wild-type and the 'bud-flowering' type by histological analyses. Analysis of epidermal cell surface structure of vegetative tissues and perianth organs using scanning electron microscopy revealed that in wild-type flowers the outer whorls of colored organs may be identified as sepals, while the inner ones may be identified as petals. In the 'bud-flowering' type, two whorls of sepals are directly followed by the gynoecium. Both, petals and stamens, are completely missing in this flower type. The uppermost whorl of green leaves represents bracts in both flower types. In addition, two MADS-box genes (homologs of AP3/DEF and SEP1/2) were identified in C. vulgaris using RACE-PCR. Expression analysis by qRT-PCR was conducted for both genes in leaves, bracts, sepals and petals. These experiments revealed an expression pattern supporting the organ classification based on morphological characteristics. CONCLUSIONS: Organ identity in both wild-type and 'bud flowering' C. vulgaris was clarified using a combination of microscopic and molecular methods. Our results for bract, sepal and petal organ identity are supported by the 'ABCDE model'. However, loss of stamens in the 'bud-flowering' phenotype is an exceptional flower organ modification that cannot be explained by modified spatial expression of known organ identity genes. PMID- 20003431 TI - Aqueous extracts from dietary supplements influence the production of inflammatory cytokines in immortalized and primary T lymphocytes. AB - BACKGROUND: Congaplex and Immuplex are dietary supplements that have been traditionally used to support immune system function. The purpose of these experiments was to determine whether Congaplex and Immuplex affect immune function using primary and immortalized T lymphocytes. METHODS: Immortalized CEM and Jurkat T lymphocytes and primary peripheral mononuclear blood cells (PBMCs) were treated with the aqueous extracts from Congaplex and Immuplex to determine the effects of these products on cytokine production in activated T lymphocytes. RESULTS: Congaplex enhanced phytohemagglutinin/phorbol 12-myristate 13-acetate (PHA/PMA) stimulation of both CEM and Jurkat cells as measured by the production of cytokines, while Immuplex suppressed PHA/PMA-induced production of cytokines, with the exception of interleukin (IL)-8 which was enhanced by Immuplex. In vitro treatment of PBMCs from 10 healthy donors with Congaplex or Immuplex decreased PHA-stimulated production of interferon (IFN)-gamma but increased the production of IL-13. CONCLUSIONS: While the effects of Congaplex and Immuplex differed in these two models, these data demonstrate that the aqueous extracts from these two dietary supplements can affect the inflammatory response of T lymphocytes. PMID- 20003432 TI - Helicobacter pylori lipopolysaccharide modification, Lewis antigen expression, and gastric colonization are cholesterol-dependent. AB - BACKGROUND: Helicobacter pylori specifically takes up cholesterol and incorporates it into the bacterial membrane, yet little is currently known about cholesterol's physiological roles. We compared phenotypes and in vivo colonization ability of H. pylori grown in a defined, serum-free growth medium, F12 with 1 mg/ml albumin containing 0 to 50 mug/ml cholesterol. RESULTS: While doubling times were largely unaffected by cholesterol, other overt phenotypic changes were observed. H. pylori strain SS1 grown in defined medium with cholesterol successfully colonized the stomach of gerbils, whereas SS1 grown without cholesterol failed to colonize. H. pylori lipopolysaccharide often displays Lewis X and/or Y antigens. Expression of these antigens measured by whole-cell ELISA was markedly enhanced in response to growth of strain SS1, 26695, or G27 in cholesterol. In addition, electrophoretic analysis of lipopolysaccharide in wild type G27 and in mutants lacking the O-chain revealed structural changes within the oligosaccharide core/lipid A moieties. These responses in Lewis antigen levels and in lipopolysaccharide profiles to cholesterol availability were highly specific, because no changes took place when cholesterol was substituted by beta-sitosterol or bile salts. Disruption of the genes encoding cholesterol alpha-glucosyltransferase or lipid A phosphoethanolamine transferase had no effect on Lewis expression, nor on lipopolysaccharide profiles, nor on the cholesterol responsiveness of these properties. Disruption of the lipid A 1-phosphatase gene eliminated the effect of cholesterol on lipopolysaccharide profiles but not its effect on Lewis expression. CONCLUSIONS: Together these results suggest that cholesterol depletion leads to aberrant forms of LPS that are dependent upon dephosphorylation of lipid A at the 1-position. A tentative model for the observed effects of cholesterol is discussed in which sequential steps of lipopolysaccharide biogenesis and, independently, presentation of Lewis antigen at the cell surface, depend upon membrane composition. These new findings demonstrate that cholesterol availability permits H. pylori to modify its cell envelope in ways that can impact colonization of host tissue in vivo. PMID- 20003433 TI - TransportTP: a two-phase classification approach for membrane transporter prediction and characterization. AB - BACKGROUND: Membrane transporters play crucial roles in living cells. Experimental characterization of transporters is costly and time-consuming. Current computational methods for transporter characterization still require extensive curation efforts, especially for eukaryotic organisms. We developed a novel genome-scale transporter prediction and characterization system called TransportTP that combined homology-based and machine learning methods in a two phase classification approach. First, traditional homology methods were employed to predict novel transporters based on sequence similarity to known classified proteins in the Transporter Classification Database (TCDB). Second, machine learning methods were used to integrate a variety of features to refine the initial predictions. A set of rules based on transporter features was developed by machine learning using well-curated proteomes as guides. RESULTS: In a cross validation using the yeast proteome for training and the proteomes of ten other organisms for testing, TransportTP achieved an equivalent recall and precision of 81.8%, based on TransportDB, a manually annotated transporter database. In an independent test using the Arabidopsis proteome for training and four recently sequenced plant proteomes for testing, it achieved a recall of 74.6% and a precision of 73.4%, according to our manual curation. CONCLUSIONS: TransportTP is the most effective tool for eukaryotic transporter characterization up to date. PMID- 20003434 TI - Paediatricians knowledge, attitudes, and practices regarding immunizations for infants in Italy. AB - BACKGROUND: The purpose of this study was to investigate whether paediatricians have appropriate knowledge, attitudes, and behaviours regarding vaccinations for infants in Italy. METHODS: A random sample of 500 paediatricians received a self administered anonymous questionnaire covering demographic and professional characteristics; knowledge about the mandatory, recommended, and not indicated vaccinations for infants; attitudes about vaccinations for infants; behaviour regarding current administration or willingness to administer mandatory or recommended vaccinations for infants and immunization education programs of the parents. RESULTS: Only 42.3% paediatricians knew all recommended vaccinations for infants and this knowledge was significantly higher in females, in those who worked a higher number of hours for week, and in those who use guidelines for immunization practice. Only 10.3% had a very favourable attitude towards the utility of the recommended vaccinations for infants and this was significantly higher in those who administered recommended vaccinations for infants. A large proportion (82.7%) of paediatricians routinely informed the parents about the recommended vaccinations for infants and this appropriate behaviour was significantly higher among younger, in those with a higher number of years in practice, and in those who administered the recommended vaccinations for infants. CONCLUSION: Training and educational interventions are needed in order to improve knowledge, attitudes, and behaviours regarding vaccinations for infants among paediatricians. PMID- 20003435 TI - A new two-phase dimeticone pediculicide shows high efficacy in a comparative bioassay. AB - BACKGROUND: Dimeticones kill head lice by physical means. Here we assessed in a comparative bioassay the ex vivo efficacy of "NYDA sensitiv", a new two-phase dimeticone-based pediculicide similar to a product established on the market, but without fragrances. METHODS: We compared efficacy of the new product to a positive dimeticone control group, a sample of four other insecticidal and natural head lice products marketed in Germany, and an untreated control. In a bioassay, lice were exposed ex vivo to products and examined for activity for up to 24 hours, following a standard protocol. RESULTS: After 6 and 24 hours, 13.7 and 88.5% of untreated control lice did not show major vital signs. In contrast, no lice showed major vital signs 5 minutes after treatment with the new product or the control dimeticone group (NYDA). This effect persisted at all observation points (100% efficacy). Efficacy of 0.5% permethrin (Infectopedicul) ranged between 76 and 96% in evaluations between 5 min and 6 hours. All lice treated with a coconut-based compound (mosquito Lauseshampoo) did not show major vital signs after 5 min, but mortality was only 58% after one hour. Pyrethrum extract (Goldgeist forte) showed an efficacy of 22 - 52% between 5 min and 3 hours after treatment; after 6 hours, 76% of lice were judged dead. An oxyphthirine-based compound (Liberalice DUO LP-PRO) killed 22 - 54% of lice in the first 6 hours. CONCLUSIONS: The two-phase dimeticone compound NYDA sensitiv is highly efficacious. The removal of fragrances as compared to an established dimeticone product did not affect in vitro efficacy. PMID- 20003436 TI - SWiss Atorvastatin and interferon Beta-1b trial In Multiple Sclerosis (SWABIMS)- rationale, design and methodology. AB - BACKGROUND: Statins have anti-inflammatory and immunomodulatory properties in addition to their lipid-lowering effects. Currently, the effects of statins on multiple sclerosis are still controversial. Therefore, randomized clinical trials are needed to provide better evidence on the therapeutic potential of statins in multiple sclerosis. The SWiss Atorvastatin and Interferon Beta-1b trial in Multiple Sclerosis (SWABIMS) evaluates the efficacy, safety and tolerability of atorvastatin 40 mg per os daily and subcutaneous interferon beta-1b every other day compared to monotherapy with subcutaneous interferon beta-1b every other day in patients with relapsing-remitting multiple sclerosis. METHODS/DESIGN: SWABIMS is a multi-centre, randomized, parallel-group, rater-blinded, Phase IIb-study conducted in eight hospitals in Switzerland. 80 treatment naive patients with relapsing-remitting forms of multiple sclerosis will receive subcutaneous interferon beta-1b for three months. Afterwards, they are randomized into two equal-sized parallel arms, receiving atorvastatin 40 mg/d or not in addition to interferon beta-1b for another 12 months. Disease activity measured by the proportion of patients with new T2 lesions is the primary endpoint. DISCUSSION: SWABIMS is designed to give further information about the therapeutic effect of atorvastatin 40 mg per os daily as add-on therapy to interferon beta-1b in patients with relapsing-remitting multiple sclerosis. Furthermore important safety and tolerability data will be generated. TRIAL REGISTRATION: http://www.clinicaltrials.gov. Identifier: NCT00942591; Swissmedic reference number: 2005DR2119. PMID- 20003437 TI - Peripheral non-viral MIDGE vector-driven delivery of beta-endorphin in inflammatory pain. AB - BACKGROUND: Leukocytes infiltrating inflamed tissue produce and release opioid peptides such as beta-endorphin, which activate opioid receptors on peripheral terminals of sensory nerves resulting in analgesia. Gene therapy is an attractive strategy to enhance continuous production of endogenous opioids. However, classical viral and plasmid vectors for gene delivery are hampered by immunogenicity, recombination, oncogene activation, anti-bacterial antibody production or changes in physiological gene expression. Non-viral, non-plasmid minimalistic, immunologically defined gene expression (MIDGE) vectors may overcome these problems as they carry only elements needed for gene transfer. Here, we investigated the effects of a nuclear localization sequence (NLS) coupled MIDGE encoding the beta-endorphin precursor proopiomelanocortin (POMC) on complete Freund's adjuvant-induced inflammatory pain in rats. RESULTS: POMC-MIDGE NLS injected into inflamed paws appeared to be taken up by leukocytes resulting in higher concentrations of beta-endorphin in these cells. POMC-MIDGE-NLS treatment reversed enhanced mechanical sensitivity compared with control MIDGE NLS. However, both effects were moderate, not always statistically significant or directly correlated with each other. Also, the anti-hyperalgesic actions could not be increased by enhancing beta-endorphin secretion or by modifying POMC-MIDGE NLS to code for multiple copies of beta-endorphin. CONCLUSION: Although MIDGE vectors circumvent side-effects associated with classical viral and plasmid vectors, the current POMC-MIDGE-NLS did not result in reliable analgesic effectiveness in our pain model. This was possibly associated with insufficient and variable efficacy in transfection and/or beta-endorphin production. Our data point at the importance of the reproducibility of gene therapy strategies for the control of chronic pain. PMID- 20003438 TI - Evolution of antibody landscape and viral envelope escape in an HIV-1 CRF02_AG infected patient with 4E10-like antibodies. AB - BACKGROUND: A minority of HIV-1 infected individuals develop broad cross neutralizing (BCN) plasma antibodies that are capable of neutralizing a spectrum of virus variants belonging to different HIV-1 clades. The aim of this study was to identify the targeted epitopes of an individual with BCN plasma antibodies, referred to as ITM4, using peptide phage display. This study also aimed to use the selected mimotopes as tools to unravel the evolution of the antibody landscape and the viral envelope escape which may provide us with new insights for vaccine design. RESULTS: This study led us to identify ITM4 plasma antibodies directed to the 4E10 epitope located in the gp41 membrane-proximal external region (MPER). Analysis of antibody specificities revealed unusual immunogenic properties of the ITM4 viral envelope, as not only the V3 loop and the gp41 MPER but also the C1 and lentivirus lytic peptide 2 (LLP2) region seem to be targets of the immune system. The 4E10-like antibodies are consistently elicited during the 6-year follow up period. HIV-1 ITM4 pseudoviruses showed an increasing resistance over time to MPER monoclonal antibodies 4E10 and 2F5, although no changes are found in the critical positions of the epitope. Neutralization of COT6.15 (subtype C; 4E10-sensitive) pseudoviruses with alanine substitutions in the MPER region indicated an overlapping specificity of the 4E10 monoclonal antibody and the ITM4 follow up plasma. Moreover the 4E10-like antibodies of ITM4 contribute to the BCN capacity of the plasma. CONCLUSIONS: Using ITM4 BCN plasma and peptide phage display technology, we have identified a patient with 4E10-like BCN antibodies. Our results indicate that the elicited 4E10-like antibodies play a role in virus neutralization. The viral RNA was isolated at different time points and the ITM4 envelope sequence analysis of both early (4E10-sensitive) and late (4E10-resistant) viruses suggest that other regions in the envelope, outside the MPER region, contribute to the accessibility and sensitivity of the 4E10 epitope. Including ITM4 specific HIV-1 Env properties in vaccine strategies may be a promising approach. PMID- 20003439 TI - Detecting coordinated regulation of multi-protein complexes using logic analysis of gene expression. AB - BACKGROUND: Many of the functional units in cells are multi-protein complexes such as RNA polymerase, the ribosome, and the proteasome. For such units to work together, one might expect a high level of regulation to enable co-appearance or repression of sets of complexes at the required time. However, this type of coordinated regulation between whole complexes is difficult to detect by existing methods for analyzing mRNA co-expression. We propose a new methodology that is able to detect such higher order relationships. RESULTS: We detect coordinated regulation of multiple protein complexes using logic analysis of gene expression data. Specifically, we identify gene triplets composed of genes whose expression profiles are found to be related by various types of logic functions. In order to focus on complexes, we associate the members of a gene triplet with the distinct protein complexes to which they belong. In this way, we identify complexes related by specific kinds of regulatory relationships. For example, we may find that the transcription of complex C is increased only if the transcription of both complex A AND complex B is repressed. We identify hundreds of examples of coordinated regulation among complexes under various stress conditions. Many of these examples involve the ribosome. Some of our examples have been previously identified in the literature, while others are novel. One notable example is the relationship between the transcription of the ribosome, RNA polymerase and mannosyltransferase II, which is involved in N-linked glycan processing in the Golgi. CONCLUSIONS: The analysis proposed here focuses on relationships among triplets of genes that are not evident when genes are examined in a pairwise fashion as in typical clustering methods. By grouping gene triplets, we are able to decipher coordinated regulation among sets of three complexes. Moreover, using all triplets that involve coordinated regulation with the ribosome, we derive a large network involving this essential cellular complex. In this network we find that all multi-protein complexes that belong to the same functional class are regulated in the same direction as a group (either induced or repressed). PMID- 20003440 TI - Cross-species amplification of 36 cyprinid microsatellite loci in Phoxinus phoxinus (L.) and Scardinius erythrophthalmus (L.). AB - BACKGROUND: To conduct phylogeographic or population genetic studies, an adequate number of DNA markers for the focal species are required. Due to severe unavailability of genotype markers of any kind for the species Eurasian minnow (Phoxinus phoxinus L.) and rudd (Scardinius erythrophthalmus L.), we set out to attempt cross-amplification of a set of microsatellite loci from related species. FINDINGS: We tested 36 cyprinid microsatellite loci for cross-species amplification in minnow and rudd. Fifteen species-locus combinations produced amplifications in minnow, seven being polymorphic, while 18 combinations amplified in rudd, nine of these being polymorphic. CONCLUSIONS: The positive cross-species amplifications present potential contributions to the establishment of genetic marker sets for population genetics studies of the two focal species. PMID- 20003441 TI - The Golgi alpha-1,6 mannosyltransferase KlOch1p of Kluyveromyces lactis is required for Ca2+/calmodulin-based signaling and for proper mitochondrial functionality. AB - BACKGROUND: Protein N-glycosylation is a relevant metabolic pathway in eukaryotes and plays key roles in cell processes. In yeasts, outer chain branching is initiated in the Golgi apparatus by the alpha-1,6-mannosyltransferase Och1p. RESULTS: Here we report that, in Kluyveromyces lactis, this glycosyltransferase is also required to maintain functional mitochondria and calcium homeostasis. Cells carrying a mutation in KlOCH1 gene showed altered mitochondrial morphology, increased accumulation of ROS and reduced expression of calcium signalling genes such as calmodulin and calcineurin. Intracellular calcium concentration was also reduced in the mutant cells with respect to the wild type counterparts.Phenotypes that occur in cells lacking the alpha-1,6-mannosyltransferase, including oxidative stress and impaired mitochondria functionality, were suppressed by increased dosage of KlCmd1p. This, in turn, acts through the action of calcineurin. CONCLUSIONS: Proper functioning of the alpha-1,6-mannosyltransferase in the N-glycosylation pathway of K. lactis is required for maintaining normal calcium homeostasis; this is necessary for physiological mitochondria dynamics and functionality. PMID- 20003442 TI - Critical assessment of sequence-based protein-protein interaction prediction methods that do not require homologous protein sequences. AB - BACKGROUND: Protein-protein interactions underlie many important biological processes. Computational prediction methods can nicely complement experimental approaches for identifying protein-protein interactions. Recently, a unique category of sequence-based prediction methods has been put forward--unique in the sense that it does not require homologous protein sequences. This enables it to be universally applicable to all protein sequences unlike many of previous sequence-based prediction methods. If effective as claimed, these new sequence based, universally applicable prediction methods would have far-reaching utilities in many areas of biology research. RESULTS: Upon close survey, I realized that many of these new methods were ill-tested. In addition, newer methods were often published without performance comparison with previous ones. Thus, it is not clear how good they are and whether there are significant performance differences among them. In this study, I have implemented and thoroughly tested 4 different methods on large-scale, non-redundant data sets. It reveals several important points. First, significant performance differences are noted among different methods. Second, data sets typically used for training prediction methods appear significantly biased, limiting the general applicability of prediction methods trained with them. Third, there is still ample room for further developments. In addition, my analysis illustrates the importance of complementary performance measures coupled with right-sized data sets for meaningful benchmark tests. CONCLUSIONS: The current study reveals the potentials and limits of the new category of sequence-based protein-protein interaction prediction methods, which in turn provides a firm ground for future endeavours in this important area of contemporary bioinformatics. PMID- 20003443 TI - Lack of functional and expression homology between human and mouse aldo-keto reductase 1C enzymes: implications for modelling human cancers. AB - BACKGROUND: Over recent years, enzymes of the aldo-keto reductase (AKR) 1C subfamily have been implicated in the progression of prostate, breast, endometrial and leukemic cancers. This is due to the ability of AKR1C enzymes to modify androgens, estrogens, progesterone and prostaglandins (PGs) in a tissue specific manner, regulating the activity of nuclear receptors and other downstream effects. Evidence supporting a role for AKR1C enzymes in cancer derives mostly from studies with isolated primary cells from patients or immortalized cell lines. Mice are ideal organisms for in vivo studies, using knock-out or over-expression strains. However, the functional conservation of AKR1C enzymes between human and mice has yet to be described. RESULTS: In this study, we have characterized and compared the four human (AKR1C1,-1C2, -1C3 and 1C4) and the eight murine (AKR1C6, -1C12, -1C13, -1C14, -1C18, -1C19, -1C20 and 1C21) isoforms in their phylogeny, substrate preference and tissue distribution. We have found divergent evolution between human and murine AKR1C enzymes that was reflected by differing substrate preference. Murine enzymes did not perform the 11beta-ketoreduction of prostaglandin (PG) D2, an activity specific to human AKR1C3 and important in promoting leukemic cell survival. Instead, murine AKR1C6 was able to perform the 9-ketoreduction of PGE2, an activity absent amongst human isoforms. Nevertheless, reduction of the key steroids androstenedione, 5alpha dihydrotestosterone, progesterone and estrone was found in murine isoforms. However, unlike humans, no AKR1C isoforms were detected in murine prostate, testes, uterus and haemopoietic progenitors. CONCLUSIONS: This study exposes significant lack of phylogenetic and functional homology between human and murine AKR1C enzymes. Therefore, we conclude that mice are not suitable to model the role of AKR1C in human cancers and leukemia. PMID- 20003444 TI - Application of hybridization control probe to increase accuracy on ligation detection or minisequencing diagnostic microarrays. AB - BACKGROUND: Nucleic acid detection based on ligation reaction or single nucleotide extension of ssDNA probes followed by tag microarray hybridization provides an accurate and sensitive detection tool for various diagnostic purposes. Since microarray quality is crucial for reliable detection, these methods can benefit from correcting for microarray artefacts using specifically adapted techniques. FINDINGS: Here we demonstrate the application of a per-spot hybridization control oligonucleotide probe and a novel way of computing normalization for tag array data. The method takes into account the absolute value of the detection probe signal and the variability in the control probe signal to significantly alleviate problems caused by artefacts and noise on low quality microarrays. CONCLUSIONS: Diagnostic microarray platforms require experimental and computational tools to enable efficient correction of array artefacts. The techniques presented here improve the signal to noise ratio and help in determining true positives with better statistical significance and in allowing the use of arrays with poor quality that would otherwise be discarded. PMID- 20003445 TI - A new deep branch of eurasian mtDNA macrohaplogroup M reveals additional complexity regarding the settlement of Madagascar. AB - BACKGROUND: Current models propose that mitochondrial DNA macrohaplogroups M and N evolved from haplogroup L3 soon after modern humans left Africa. Increasingly, however, analysis of isolated populations is filling in the details of, and in some cases challenging, aspects of this general model. RESULTS: Here, we present the first comprehensive study of three such isolated populations from Madagascar: the Mikea hunter-gatherers, the neighbouring Vezo fishermen, and the Merina central highlanders (n = 266). Complete mitochondrial DNA genome sequences reveal several unresolved lineages, and a new, deep branch of the out-of-Africa founder clade M has been identified. This new haplogroup, M23, has a limited global distribution, and is restricted to Madagascar and a limited range of African and Southwest Asian groups. CONCLUSIONS: The geographic distribution, phylogenetic placement and molecular age of M23 suggest that the colonization of Madagascar was more complex than previously thought. PMID- 20003446 TI - Evaluation of current knowledge, awareness and practice of spirometry among hospital -based Nigerian doctors. AB - BACKGROUND: Spirometry is a cost-effective diagnostic tool for evaluation of lung function and for case-finding in a resource-limited setting. The acceptance of this test depends on the awareness of its indications and the ability to interpret the results. No studies have assessed the knowledge of spirometry among Nigerian doctors. The aim of this study was to evaluate the current knowledge, awareness and practice of spirometry among hospital-based Nigerian doctors. METHODS: We carried out a cross-sectional survey among 321 doctors working in Nigerian hospitals between March 2008 and June 2008. Information on knowledge, awareness, practice of and barriers to spirometry were obtained using a pre tested, self-administered structured questionnaire and the data were then analysed. RESULTS: Of the 321 doctors that participated, 108 (33.6%) reported that they have good knowledge of spirometry. One hundred and ninety-five (60.7%) were aware of the importance of spirometry in aiding the diagnosis of respiratory diseases; 213(66.4%) were aware of the importance of spirometry in determining the severity of diseases. Medical school was the most common source of knowledge on spirometry (64.5%). Eighty-one (25.2%) doctors reported having a spirometer in their hospitals. Doctors having access to a spirometer used it more frequently for aiding the diagnosis of COPD (40.7% vs.27.5%) and for monitoring of asthma (18.5% vs.11.3%) than those without access to a spirometer. The doctors working in University Teaching Hospitals and Federal Medical Centres (FMC) (22.4% vs. 4.5%) and those having access to a spirometer (40.7 vs.11.3%) were very confident of interpreting spirometry results compared to those working in District and General Hospitals and without access to a spirometer. Irrespective of access to a spirometer or the type of hospital they were employed in, doctors reported that unavailability of a spirometer was the greatest barrier to its use (62.5%) followed by lack of awareness about its usefulness (17.2%). CONCLUSION: The knowledge and practice of spirometry were poor among hospital-based Nigerian doctors because of unavailability of spirometers in most hospitals. These findings have implications for further evaluation, planning and management of patient care in respiratory disease. Spirometers should be made available in all hospitals, and the knowledge of spirometry should be improved among doctors. PMID- 20003448 TI - Isolated recurrence of distal adenocarcinoma of the extrahepatic bile duct on a draining sinus scar after curative resection: case report and review of the literature. AB - BACKGROUND: Surgical resection remains the gold standard for the treatment of localized adenocarcinoma of the extrahepatic bile ducts. Yet, treatment of loco regional recurrences is not well defined. CASE PRESENTATION: We present an unusual case of distal adenocarcinoma of the extrahepatic bile ducts that was treated with surgery and relapsed two years later with a solitary recurrence on the tract of a previous Redon drain. In addition, a review of the literature on management of loco regional relapses is presented. CONCLUSIONS: The ideal management of these patients still remains undefined. Decisions are made based on clinical parameters from retrospective series, such as tumor grade, surgical margins or lymph node involvement. Prospective studies, that include molecular and genetic markers, are needed to improve patient selection and outcomes on this population. PMID- 20003447 TI - Nuclear receptor coregulator SNP discovery and impact on breast cancer risk. AB - BACKGROUND: Coregulator proteins are "master regulators", directing transcriptional and posttranscriptional regulation of many target genes, and are critical in many normal physiological processes, but also in hormone driven diseases, such as breast cancer. Little is known on how genetic changes in these genes impact disease development and progression. Thus, we set out to identify novel single nucleotide polymorphisms (SNPs) within SRC-1 (NCoA1), SRC-3 (NCoA3, AIB1), NCoR (NCoR1), and SMRT (NCoR2), and test the most promising SNPs for associations with breast cancer risk. METHODS: The identification of novel SNPs was accomplished by sequencing the coding regions of these genes in 96 apparently normal individuals (48 Caucasian Americans, 48 African Americans). To assess their association with breast cancer risk, five SNPs were genotyped in 1218 familial BRCA1/2-mutation negative breast cancer cases and 1509 controls (rs1804645, rs6094752, rs2230782, rs2076546, rs2229840). RESULTS: Through our resequencing effort, we identified 74 novel SNPs (30 in NCoR, 32 in SMRT, 10 in SRC-3, and 2 in SRC-1). Of these, 8 were found with minor allele frequency (MAF) >5% illustrating the large amount of genetic diversity yet to be discovered. The previously shown protective effect of rs2230782 in SRC-3 was strengthened (OR = 0.45 [0.21-0.98], p = 0.04). No significant associations were found with the other SNPs genotyped. CONCLUSIONS: This data illustrates the importance of coregulators, especially SRC-3, in breast cancer development and suggests that more focused studies, including functional analyses, should be conducted. PMID- 20003449 TI - Dual agarose magnetic (DAM) ChIP. AB - BACKGROUND: Chromatin immunoprecipitation (ChIP) has become a very popular technique to study epigenetic regulation because it can be used to identify proteins and protein modifications present at specific locations in chromatin. While techniques have been developed to investigate epigenetic modifications present in chromatin during a specific biological function such as transcription, they depend upon the ability of the ChIP to analyze two epitopes on the same chromatin and are generally time consuming, difficult to perform, and not very sensitive. The Dual Agarose Magnetic (DAM) ChIP procedure described here is designed to address these shortcomings. FINDINGS: Protein A agarose and protein G magnetic beads bound with different IgGs have been combined in a single Chromatin Immunoprecipitation (ChIP) assay to analyze for the presence of two epitopes on the same chromatin at the same time. This procedure has been used with non-immune rabbit IgG bound to either the agarose or beads in order to include an internal negative control for non-specific binding of chromatin. The procedure has also been used with various antibodies including those targeting RNA Polymerase II and replication protein A 70 to determine whether specific forms of modified histones are present in either transcribing or replicating forms of SV40 minichromosomes respectively. CONCLUSIONS: The DAM ChIP procedure is a rapid, simple, and sensitive technique to characterize two epitopes located in the same chromatin. It should be particularly useful for the rapid screening of epigenetic modifications present in biologically active chromatin. PMID- 20003450 TI - A proposal for a new clinical classification of chronic pancreatitis. AB - BACKGROUND: The clinical course of chronic pancreatitis is still unpredictable, which relates to the lack of the availability of a clinical classification. Therefore, patient populations cannot be compared, the course and the outcome of the disease remain undetermined in the individual patient, and treatment is not standardized. AIM: To establish a clinical classification for chronic pancreatitis which is user friendly, transparent, relevant, prognosis- as well as treatment-related and offers a frame for future disease evaluation. METHODS: Diagnostic requirements will include one clinical criterion, in combination with well defined imaging or functional abnormalities. RESULTS: A classification system consisting of three stages (A, B and C) is presented, which fulfils the above-mentioned criteria. Clinical criteria are: pain, recurrent attacks of pancreatitis, complications of chronic pancreatitis (e.g. bile duct stenosis), steatorrhea, and diabetes mellitus. Imaging criteria consist of ductal or parenchymal changes observed by ultrasonography, ERCP, CT, MRI, and/or endosonography. CONCLUSION: A new classification of chronic pancreatitis, based on combination of clinical signs, morphology and function, is presented. It is easy to handle and an instrument to study and to compare the natural course, the prognosis and treatment of patients with chronic pancreatitis. PMID- 20003451 TI - Local expression of tumor necrosis factor-receptor 1:immunoglobulin G can induce salivary gland dysfunction in a murine model of Sjogren's syndrome. AB - INTRODUCTION: Tumor necrosis factor is a pleiotropic cytokine with potent immune regulatory functions. Although tumor necrosis factor inhibitors have demonstrated great utility in treating other autoimmune diseases, such as rheumatoid arthritis, there are conflicting results in Sjogren's syndrome. The aim of this study was to assess the effect of a locally expressed tumor necrosis factor inhibitor on the salivary gland function and histopathology in an animal model of Sjogren's syndrome. METHODS: Using in vivo adeno associated viral gene transfer, we have stably expressed soluble tumor necrosis factor-receptor 1-Fc fusion protein locally in the salivary glands in the Non Obese Diabetic model of Sjogren's syndrome. Pilocarpine stimulated saliva flow was measured to address the salivary gland function and salivary glands were analyzed for focus score and cytokine profiles. Additionally, cytokines and autoantibody levels were measured in plasma. RESULTS: Local expression of tumor necrosis factor-receptor 1:immunoglobulin G fusion protein resulted in decreased saliva flow over time. While no change in lymphocytic infiltrates or autoantibody levels was detected, statistically significant increased levels of tumor growth factor-beta1 and decreased levels of interleukin-5, interleukin-12p70 and interleukin -17 were detected in the salivary glands. In contrast, plasma levels showed significantly decreased levels of tumor growth factor-beta1 and increased levels of interleukin 4, interferon-gamma, interleukin-10 and interleukin-12p70. CONCLUSIONS: Our findings suggest that expression of tumor necrosis factor inhibitors in the salivary gland can have a negative effect on salivary gland function and that other cytokines should be explored as points for therapeutic intervention in Sjogren's syndrome. PMID- 20003452 TI - Reduced transcription of TCOF1 in adult cells of Treacher Collins syndrome patients. AB - BACKGROUND: Treacher Collins syndrome (TCS) is an autosomal dominant craniofacial disorder caused by frameshift deletions or duplications in the TCOF1 gene. These mutations cause premature termination codons, which are predicted to lead to mRNA degradation by nonsense mediated mRNA decay (NMD). Haploinsufficiency of the gene product (treacle) during embryonic development is the proposed molecular mechanism underlying TCS. However, it is still unknown if TCOF1 expression levels are decreased in post-embryonic human cells. METHODS: We have estimated TCOF1 transcript levels through real time PCR in mRNA obtained from leucocytes and mesenchymal cells of TCS patients (n = 23) and controls (n = 18). Mutational screening and analysis of NMD were performed by direct sequencing of gDNA and cDNA, respectively. RESULTS: All the 23 patients had typical clinical features of the syndrome and pathogenic mutations were detected in 19 of them. We demonstrated that the expression level of TCOF1 is 18-31% lower in patients than in controls (p < 0.05), even if we exclude the patients in whom we did not detect the pathogenic mutation. We also observed that the mutant allele is usually less abundant than the wild type one in mesenchymal cells. CONCLUSIONS: This is the first study to report decreased expression levels of TCOF1 in TCS adult human cells, but it is still unknown if this finding is associated to any phenotype in adulthood. In addition, as we demonstrated that alleles harboring the pathogenic mutations have lower expression, we herein corroborate the current hypothesis of NMD of the mutant transcript as the explanation for diminished levels of TCOF1 expression. Further, considering that TCOF1 deficiency in adult cells could be associated to pathologic clinical findings, it will be important to verify if TCS patients have an impairment in adult stem cell properties, as this can reduce the efficiency of plastic surgery results during rehabilitation of these patients. PMID- 20003453 TI - Adaptive behavior of neighboring neurons during adaptation-induced plasticity of orientation tuning in VI. AB - BACKGROUND: Sensory neurons display transient changes of their response properties following prolonged exposure to an appropriate stimulus (adaptation). In adult cat primary visual cortex, orientation-selective neurons shift their preferred orientation after being adapted to a non-preferred orientation. The direction of those shifts, towards (attractive) or away (repulsive) from the adapter depends mostly on adaptation duration. How the adaptive behavior of a neuron is related to that of its neighbors remains unclear. RESULTS: Here we show that in most cases (75%), cells shift their preferred orientation in the same direction as their neighbors. We also found that cells shifting preferred orientation differently from their neighbors (25%) display three interesting properties: (i) larger variance of absolute shift amplitude, (ii) wider tuning bandwidth and (iii) larger range of preferred orientations among the cluster of cells. Several response properties of V1 neurons depend on their location within the cortical orientation map. Our results suggest that recording sites with both attractive and repulsive shifts following adaptation may be located in close proximity to iso-orientation domain boundaries or pinwheel centers. Indeed, those regions have a more diverse orientation distribution of local inputs that could account for the three properties above. On the other hand, sites with all cells shifting their preferred orientation in the same direction could be located within iso-orientation domains. CONCLUSIONS: Our results suggest that the direction and amplitude of orientation preference shifts in V1 depend on location within the orientation map. This anisotropy of adaptation-induced plasticity, comparable to that of the visual cortex itself, could have important implications for our understanding of visual adaptation at the psychophysical level. PMID- 20003454 TI - The role of beta-lactamase-producing-bacteria in mixed infections. AB - Beta-lactamase-producing bacteria (BLPB) can play an important role in polymicrobial infections. They can have a direct pathogenic impact in causing the infection as well as an indirect effect through their ability to produce the enzyme beta-lactamase. BLPB may not only survive penicillin therapy but can also, as was demonstrated in in vitro and in vivo studies, protect other penicillin susceptible bacteria from penicillin by releasing the free enzyme into their environment. This phenomenon occurs in upper respiratory tract, skin, soft tissue, surgical and other infections. The clinical, in vitro, and in vivo evidence supporting the role of these organisms in the increased failure rate of penicillin in eradication of these infections and the implication of that increased rate on the management of infections is discussed. PMID- 20003455 TI - Characterization of the transcripts and protein isoforms for cytoplasmic polyadenylation element binding protein-3 (CPEB3) in the mouse retina. AB - BACKGROUND: Cytoplasmic polyadenylation element binding proteins (CPEBs) regulate translation by binding to regulatory motifs of defined mRNA targets. This translational mechanism has been shown to play a critical role in oocyte maturation, early development, and memory formation in the hippocampus. Little is known about the presence or functions of CPEBs in the retina. The purpose of the current study is to investigate the alternative splicing isoforms of a particular CPEB, CPEB3, based on current databases, and to characterize the expression of CPEB3 in the retina. RESULTS: In this study, we have characterized CPEB3, whose putative role is to regulate the translation of GluR2 mRNA. We identify the presence of multiple alternative splicing isoforms of CPEB3 transcripts and proteins in the current databases. We report the presence of eight alternative splicing patterns of CPEB3, including a novel one, in the mouse retina. All but one of the patterns appear to be ubiquitous in 13 types of tissue examined. The relative abundance of the patterns in the retina is demonstrated. Experimentally, we show that CPEB3 expression is increased in a time-dependent manner during the course of postnatal development, and CPEB3 is localized mostly in the inner retina, including retinal ganglion cells. CONCLUSION: The level of CPEB3 was up regulated in the retina during development. The presence of multiple CPEB3 isoforms indicates remarkable complexity in the regulation and function of CPEB3. PMID- 20003456 TI - Biochemical evidence for the tyrosine involvement in cationic intermediate stabilization in mouse beta-carotene 15, 15'-monooxygenase. AB - BACKGROUND: beta-carotene 15,15'-monooxygenase (BCMO1) catalyzes the crucial first step in vitamin A biosynthesis in animals. We wished to explore the possibility that a carbocation intermediate is formed during the cleavage reaction of BCMO1, as is seen for many isoprenoid biosynthesis enzymes, and to determine which residues in the substrate binding cleft are necessary for catalytic and substrate binding activity. To test this hypothesis, we replaced substrate cleft aromatic and acidic residues by site-directed mutagenesis. Enzymatic activity was measured in vitro using His-tag purified proteins and in vivo in a beta-carotene-accumulating E. coli system. RESULTS: Our assays show that mutation of either Y235 or Y326 to leucine (no cation-pi stabilization) significantly impairs the catalytic activity of the enzyme. Moreover, mutation of Y326 to glutamine (predicted to destabilize a putative carbocation) almost eliminates activity (9.3% of wt activity). However, replacement of these same tyrosines with phenylalanine or tryptophan does not significantly impair activity, indicating that aromaticity at these residues is crucial. Mutations of two other aromatic residues in the binding cleft of BCMO1, F51 and W454, to either another aromatic residue or to leucine do not influence the catalytic activity of the enzyme. Our ab initio model of BCMO1 with beta-carotene mounted supports a mechanism involving cation-pi stabilization by Y235 and Y326. CONCLUSIONS: Our data are consistent with the formation of a substrate carbocation intermediate and cation-pi stabilization of this intermediate by two aromatic residues in the substrate-binding cleft of BCMO1. PMID- 20003457 TI - Hepatic profile analyses of tipranavir in Phase II and III clinical trials. AB - BACKGROUND: The risk and course of serum transaminase elevations (TEs) and clinical hepatic serious adverse event (SAE) development in ritonavir-boosted tipranavir (TPV/r) 500/200 mg BID recipients, who also received additional combination antiretroviral treatment agents in clinical trials (TPV/r-based cART), was determined. METHODS: Aggregated transaminase and hepatic SAE data through 96 weeks of TPV/r-based cART from five Phase IIb/III trials were analyzed. Patients were categorized by the presence or absence of underlying liver disease (+LD or -LD). Kaplan-Meier (K-M) probability estimates for time-to first US National Institutes of Health, Division of AIDS (DAIDS) Grade 3/4 TE and clinical hepatic SAE were determined and clinical actions/outcomes evaluated. Risk factors for DAIDS Grade 3/4 TE were identified through multivariate Cox regression statistical modeling. RESULTS: Grade 3/4 TEs occurred in 144/1299 (11.1%) patients; 123/144 (85%) of these were asymptomatic; 84% of these patients only temporarily interrupted treatment or continued, with transaminase levels returning to Grade < or = 2. At 96 weeks of study treatment, the incidence of Grade 3/4 TEs was higher among the +LD (16.8%) than among the -LD (10.1%) patients. K-M analysis revealed an incremental risk for developing DAIDS Grade 3/4 TEs; risk was greatest through 24 weeks (6.1%), and decreasing thereafter (>24-48 weeks: 3.4%, >48 weeks-72 weeks: 2.0%, >72-96 weeks: 2.2%), and higher in +LD than -LD patients at each 24-week interval. Treatment with TPV/r, co infection with hepatitis B and/or C, DAIDS grade >1 TE and CD4+ > 200 cells/mm3 at baseline were found to be independent risk factors for development of DAIDS Grade 3/4 TE; the hazard ratios (HR) were 2.8, 2.0, 2.1 and 1.5, respectively. Four of the 144 (2.7%) patients with Grade 3/4 TEs developed hepatic SAEs; overall, 14/1299 (1.1%) patients had hepatic SAEs including six with hepatic failure (0.5%). The K-M risk of developing hepatic SAEs through 96 weeks was 1.4%; highest risk was observed during the first 24 weeks and decreased thereafter; the risk was similar between +LD and -LD patients for the first 24 weeks (0.6% and 0.5%, respectively) and was higher for +LD patients, thereafter. CONCLUSION: Through 96 weeks of TPV/r-based cART, DAIDS Grade 3/4 TEs and hepatic SAEs occurred in approximately 11% and 1% of TPV/r patients, respectively; most (84%) had no significant clinical implications and were managed without permanent treatment discontinuation. Among the 14 patients with hepatic SAE, 6 experienced hepatic failure (0.5%); these patients had profound immunosuppression and the rate appears higher among hepatitis co-infected patients. The overall probability of experiencing a hepatic SAE in this patient cohort was 1.4% through 96 weeks of treatment. Independent risk factors for DAIDS Grade 3/4 TEs include TPV/r treatment, co-infection with hepatitis B and/or C, DAIDS grade >1 TE and CD4+ > 200 cells/mm3 at baseline. TRIAL REGISTRATION: US-NIH Trial registration number: NCT00144170. PMID- 20003458 TI - Web-based interrogation of gene expression signatures using EXALT. AB - BACKGROUND: Widespread use of high-throughput techniques such as microarrays to monitor gene expression levels has resulted in an explosive growth of data sets in public domains. Integration and exploration of these complex and heterogeneous data have become a major challenge. RESULTS: The EXALT (EXpression signature AnaLysis Tool) online program enables meta-analysis of gene expression profiles derived from publically accessible sources. Searches can be executed online against two large databases currently containing more than 28,000 gene expression signatures derived from GEO (Gene Expression Omnibus) and published expression profiles of human cancer. Comparisons among gene expression signatures can be performed with homology analysis and co-expression analysis. Results can be visualized instantly in a plot or a heat map. Three typical use cases are illustrated. CONCLUSIONS: The EXALT online program is uniquely suited for discovering relationships among transcriptional profiles and searching gene expression patterns derived from diverse physiological and pathological settings. The EXALT online program is freely available for non-commercial users from http://seq.mc.vanderbilt.edu/exalt/. PMID- 20003460 TI - Spatial autocorrelation analysis of health care hotspots in Taiwan in 2006. AB - BACKGROUND: Spatial analytical techniques and models are often used in epidemiology to identify spatial anomalies (hotspots) in disease regions. These analytical approaches can be used to not only identify the location of such hotspots, but also their spatial patterns. METHODS: In this study, we utilize spatial autocorrelation methodologies, including Global Moran's I and Local Getis Ord statistics, to describe and map spatial clusters, and areas in which these are situated, for the 20 leading causes of death in Taiwan. In addition, we use the fit to a logistic regression model to test the characteristics of similarity and dissimilarity by gender. RESULTS: Gender is compared in efforts to formulate the common spatial risk. The mean found by local spatial autocorrelation analysis is utilized to identify spatial cluster patterns. There is naturally great interest in discovering the relationship between the leading causes of death and well-documented spatial risk factors. For example, in Taiwan, we found the geographical distribution of clusters where there is a prevalence of tuberculosis to closely correspond to the location of aboriginal townships. CONCLUSIONS: Cluster mapping helps to clarify issues such as the spatial aspects of both internal and external correlations for leading health care events. This is of great aid in assessing spatial risk factors, which in turn facilitates the planning of the most advantageous types of health care policies and implementation of effective health care services. PMID- 20003461 TI - A cross-sectional study of Victorian Mobile Intensive Care Ambulance Paramedics knowledge of the Valsalva Manoeuvre. AB - BACKGROUND: The Valsalva Manoeuvre (VM) is a primary measure for terminating haemodynamically stable supraventricular tachycardia (SVT) in the emergency care setting. The clinical use and termination success of the VM in the prehospital setting has not been investigated to date. The objective of this study was to determine Melbourne Mobile Intensive Care Ambulance (MICA) Paramedic knowledge of the VM, and to compare this understanding with an evidence-based model of VM performance. METHODS: A cross-sectional study in the form of a face-to-face interview was used to determine Melbourne MICA Paramedic understanding of VM instruction between January and February, 2008. The results were then compared with an evidence-based model of VM performance to ascertain compliance with the three criteria of position, pressure and duration. Ethics approval was granted. RESULTS: There were 28 participants (60.9%) who elected a form of supine posturing, some 23 participants (50%) selected the syringe method of pressure generation, with 16 participants (34.8%) selecting the "as long as you can" option for duration. On comparison, one out of 46 MICA Paramedics correctly identified the three evidence-based criteria. CONCLUSIONS: The formal education of Melbourne's MICA Paramedics would benefit from the introduction of an evidence based model of VM performance, which would impact positively on patient care and may improve reversion success in the prehospital setting. The results of this study also demonstrate that an opportunity exists to promote the evidence-based VM criteria across the primary emergency care field. PMID- 20003459 TI - 2-Deoxy-D-glucose enhances TRAIL-induced apoptosis in human melanoma cells through XBP-1-mediated up-regulation of TRAIL-R2. AB - BACKGROUND: Past studies have shown that sensitivity of melanoma cells to TRAIL induced apoptosis is largely correlated with the expression levels of TRAIL death receptors on the cell surface. However, fresh melanoma isolates and melanoma tissue sections express generally low levels of death receptors for TRAIL. The clinical potential of TRAIL in the treatment of melanoma may therefore be limited unless given with agents that increase the cell surface expression of TRAIL death receptors. 2-Deoxy-D-glucose (2-DG) is a synthetic glucose analogue that inhibits glycolysis and glycosylation and blocks cell growth. It has been in clinical evaluation for its potential use as an anticancer agent. In this study, we have examined whether 2-DG and TRAIL interact to enhance their cytotoxicity towards melanoma cells. RESULTS: 2-DG did not kill melanoma cells, but enhanced TRAIL induced apoptosis in cultured melanoma cells and fresh melanoma isolates. This was associated with increased activation of the caspase cascade and mitochondrial apoptotic pathway, and was blocked by inhibition of TRAIL-R2, and to a lesser extent, inhibition of TRAIL-R1. Treatment with 2-DG up-regulated TRAIL death receptors, in particular, TRAIL-R2, on the melanoma cell surface. Up-regulation of TRAIL-R2 was due to increased transcription that was not dependent on the transcription factors, p53 and CHOP. Instead, the IRE1 alpha and ATF6 pathways of the unfolded protein response that were activated by 2-DG appeared to be involved. Moreover, XBP-1, which is known to be transcriptionally regulated by ATF6 and functionally activated by IRE1 alpha, was found to play an important role in 2-DG-mediated transcriptional up-regulation of TRAIL-R2 in melanoma cells. CONCLUSION: These results indicate that 2-DG sensitizes human melanoma cells to TRAIL-induced apoptosis by up-regulation of TRAIL-2 via the ATF6/IRE1 alpha/XBP-1 axis of the unfolded protein response. They suggest that 2-DG is a promising agent to increase the therapeutic response to TRAIL in melanoma. PMID- 20003462 TI - The perception of the hidden curriculum on medical education: an exploratory study. AB - BACKGROUND: Major curriculum reform of undergraduate medical education occurred during the past decades in the United Kingdom (UK); however, the effects of the hidden curriculum, which influence the choice of primary care as a career, have not been sufficiently recognized. While Japan, where traditionally few institutions systematically foster primary care physicians and very few have truly embraced family medicine as their guiding discipline, has also experienced meaningful curriculum reform, the effect of the hidden curriculum is not well known. The aim of this study is to identify themes pertaining to the students' perceptions of the hidden curriculum affecting undergraduate medical education in bedside learning in Japan. METHODS: Semi-structured interviews with thematic content analysis were implemented. Undergraduate year-5 students from a Japanese medical school at a Japanese teaching hospital were recruited. Interview were planned to last between 30 to 60 minutes each, over an 8-month period in 2007. The interviewees' perceptions concerning the quality of teaching in their bedside learning and related experiences were collected and analysed thematically. RESULTS: Twenty five medical students (18 males and 7 females, mean age 25 years old) consented to participate in the interviews, and seven main themes emerged: "the perception of education as having a low priority," "the prevalence of positive/negative role models," "the persistence of hierarchy and exclusivity," "the existence of gender issues," "an overburdened medical knowledge," "human relationships with colleagues and medical team members," and "first experience from the practical wards and their patients." CONCLUSIONS: Both similarities and differences were found when comparing the results to those of previous studies in the UK. Some effects of the hidden curriculum in medical education likely exist in common between the UK and Japan, despite the differences in their demographic backgrounds, cultures and philosophies. PMID- 20003463 TI - Association between polymorphisms in DNA repair genes and survival of non-smoking female patients with lung adenocarcinoma. AB - BACKGROUND: Excision repair cross-complementing group 1 (ERCC1) and group 2 (ERCC2), and X-ray repair cross-complementing group 1 (XRCC1) proteins play important roles in the repair of DNA damage and adducts. Single nucleotide polymorphisms (SNPs) of DNA repair genes are suspected to influence treatment effect and survival of cancer patients. This study aimed to investigate the relationship between polymorphisms in ERCC2, ERCC1 and XRCC1 genes and survival of non-smoking female patients with lung adenocarcinoma. METHODS: We used polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method to evaluate SNPs in ERCC2, ERCC1 and XRCC1 genes among 257 patients. RESULTS: The overall median survival time (MST) was 13.07 months. Increasing numbers of either ERCC1 118 or XRCC1 399 variant alleles were associated with shorter survival of non-smoking female lung adenocarcinoma patients (Log-rank P < 0.001). The adjusted hazard ratios (HRs) for individuals with CT or TT genotype at ERCC1 Asn118Asn were 1.48 and 2.67 compared with those with CC genotype. For polymorphism of XRCC1 399, the HRs were 1.28 and 2.68 for GA and AA genotype. When variant alleles across both polymorphisms were combined to analysis, the increasing number of variant alleles was associated with decreasing overall survival. Using the stepwise Cox regression analysis, we found that the polymorphisms in ERCC1 and XRCC1, tumor stage and chemotherapy or radiotherapy status independently predicted overall survival of non-smoking female patients with lung adenocarcinoma. CONCLUSIONS: Genetic polymorphisms in ERCC1 and XRCC1 genes might be prognostic factors in non-smoking female patients with lung adenocarcinoma. PMID- 20003464 TI - A changing picture of shigellosis in southern Vietnam: shifting species dominance, antimicrobial susceptibility and clinical presentation. AB - BACKGROUND: Shigellosis remains considerable public health problem in some developing countries. The nature of Shigellae suggests that they are highly adaptable when placed under selective pressure in a human population. This is demonstrated by variation and fluctuations in serotypes and antimicrobial resistance profile of organisms circulating in differing setting in endemic locations. Antimicrobial resistance in the genus Shigella is a constant threat, with reports of organisms in Asia being resistant to multiple antimicrobials and new generation therapies. METHODS: Here we compare microbiological, clinical and epidemiological data from patients with shigellosis over three different periods in southern Vietnam spanning 14 years. RESULTS: Our data demonstrates a shift in dominant infecting species (S. flexneri to S. sonnei) and resistance profile of the organisms circulating in southern Vietnam. We find that there was no significant variation in the syndromes associated with either S. sonnei or S. flexneri, yet the clinical features of the disease are more severe in later observations. CONCLUSIONS: Our findings show a change in clinical presentation of shigellosis in this setting, as the disease may be now more pronounced, this is concurrent with a change in antimicrobial resistance profile. These data highlight the socio-economic development of southern Vietnam and should guide future vaccine development and deployment strategies. TRIAL REGISTRATION: Current Controlled Trials ISRCTN55945881. PMID- 20003465 TI - EEVD motif of heat shock cognate protein 70 contributes to bacterial uptake by trophoblast giant cells. AB - BACKGROUND: The uptake of abortion-inducing pathogens by trophoblast giant (TG) cells is a key event in infectious abortion. However, little is known about phagocytic functions of TG cells against the pathogens. Here we show that heat shock cognate protein 70 (Hsc70) contributes to bacterial uptake by TG cells and the EEVD motif of Hsc70 plays an important role in this. METHODS: Brucella abortus and Listeria monocytogenes were used as the bacterial antigen in this study. Recombinant proteins containing tetratricopeptide repeat (TPR) domains were constructed and confirmation of the binding capacity to Hsc70 was assessed by ELISA. The recombinant TPR proteins were used for investigation of the effect of TPR proteins on bacterial uptake by TG cells and on pregnancy in mice. RESULTS: The monoclonal antibody that inhibits bacterial uptake by TG cells reacted with the EEVD motif of Hsc70. Bacterial TPR proteins bound to the C terminal of Hsc70 through its EEVD motif and this binding inhibited bacterial uptake by TG cells. Infectious abortion was also prevented by blocking the EEVD motif of Hsc70. CONCLUSIONS: Our results demonstrate that surface located Hsc70 on TG cells mediates the uptake of pathogenic bacteria and proteins containing the TPR domain inhibit the function of Hsc70 by binding to its EEVD motif. These molecules may be useful in the development of methods for preventing infectious abortion. PMID- 20003466 TI - Aural myiasis in a neonate in peninsular Malaysia. AB - Myiasis is a pathological condition in humans and animals caused by various species of dipterous larvae. Myiasis which occurs in a newborn baby is referred as neonatal myiasis. It is a rare condition and there are only a few reports to date. A case of neonatal aural myiasis in a two day old infant is reported in this paper. PMID- 20003467 TI - Lewis y antigen promotes the proliferation of ovarian carcinoma-derived RMG-I cells through the PI3K/Akt signaling pathway. AB - BACKGROUND: Lewis y antigen is difucosylated oligosaccharide and is carried by glycoconjugates at cell surface. Elevated expression of Lewis y has been found in 75% of ovarian tumor, and the high expression level is correlated to the tumor's pathological staging and prognosis. This study was to investigate the effect and the possible mechanism of Lewis y on the proliferation of human ovarian cancer cells. METHODS: We constructed a plasmid encoding alpha1,2-fucosyltransferase (alpha1,2-FT) gene and then transfected it into ovarian carcinoma-derived RMG-I cells with lowest Lewis y antigen expression level. Effect of Lewis y on cell proliferation was assessed after transfection. Changes in cell survival and signal transduction were evaluated after alpha-L-fucosidase, anti-Lewis y antibody and phosphatidylinositol 3-kinase (PI3K) inhibitor treatment. RESULTS: Our results showed that the levels of alpha1,2-FT gene and Lewis y increased significantly after transfection. The cell proliferation of ovarian carcinoma derived RMG-I cells sped up as the Lewis y antigen was increased. Both of alpha-L fucosidase and anti-Lewis y antibody inhibited the cell proliferation. The phosphorylation level of Akt was apparently elevated in Lewis y-overexpressing cells and the inhibitor of PI3K, LY294002, dramatically inhibited the growth of Lewis y-overexpressing cells. In addition, the phosphorylation intensity and difference in phosphorylation intensity between cells with different expression of alpha1,2-FT were attenuated significantly by the monoantibody to Lewis y and by the PI3K inhibitor LY294002. CONCLUSIONS: Increased expression of Lewis y antigen plays an important role in promoting cell proliferation through activating PI3K/Akt signaling pathway in ovarian carcinoma-derived RMG-I cells. Inhibition of Lewis y expression may provide a new therapeutic approach for Lewis y positive ovarian cancer. PMID- 20003468 TI - An evolved xylose transporter from Zymomonas mobilis enhances sugar transport in Escherichia coli. AB - BACKGROUND: Xylose is a second most abundant sugar component of lignocellulose besides glucose. Efficient fermentation of xylose is important for the economics of biomass-based biorefineries. However, sugar mixtures are sequentially consumed in xylose co-fermentation with glucose due to carbon catabolite repression (CCR) in microorganisms. As xylose transmembrance transport is one of the steps repressed by CCR, it is therefore of interest to develop a transporter that is less sensitive to the glucose inhibition or CCR. RESULTS: The glucose facilitator protein Glf transporter from Zymomonas mobilis, also an efficient transporter for xylose, was chosen as the target transporter for engineering to eliminate glucose inhibition on xylose uptake. The evolution of Glf transporter was carried out with a mixture of glucose and xylose in E. coli. Error-prone PCR and random deletion were employed respectively in two rounds of evolution. Aided by a high throughput screening assay using xylose analog p-nitrophenyl-beta-D xylopyranoside (pNPX) in 96-well plates, a best mutant 2-RD5 was obtained that contains several mutations, and a deletion of 134 residues (about 28% of total residues), or three fewer transmembrane sections (TMSs). It showed a 10.8-fold improvement in terms of pNPX transport activity in the presence of glucose. The fermentation performance results showed that this mutant improved xylose consumption by 42% with M9 minimal medium containing 20 g L-1 xylose only, while with the mixture sugar of xylose and glucose, 28% more glucose was consumed, but no obvious co-utilization of xylose was observed. Further glucose fed-batch experiments suggested that the intracellular metabolism of xylose was repressed by glucose. CONCLUSIONS: Through random mutagenesis and partial deletion coupled with high-throughput screening, a mutant of the Glf transporter (2-RD5) was obtained that relieved the inhibition of xylose transport by glucose. The fermentation tests revealed that 2-RD5 was advantageous in xylose and glucose uptakes, while no obvious advantage was seen for xylose co-consumption when co fermented with glucose. Further efforts could focus on reducing CCR-mediated repression of intracellular metabolism of xylose. Glf should also serve as a useful model to further exploit the molecular mechanism of xylose transport and the CCR-mediated inhibition. PMID- 20003469 TI - Association between SOD2 T-9C and MTHFR C677T polymorphisms and longevity: a study in Jordanian population. AB - BACKGROUND: Aging in animals is associated with high body oxidative stress, which might affect susceptibility and onset of age-related diseases, and the subsequent life span. Therefore, genes that modulate oxidative stress might play a role in determining longevity. In this study, we investigated whether the gene pool relevant to the SOD2-9T/C and MTHFR 677C/T polymorphisms changes as the Jordanian population ages. METHODS: Polymorphisms were genotyped in 130 elderly subjects (57 females and 73 males, mean age: 90.01 years) and 135 young control subjects (67 females and 68 males, mean age: 33.43 years). RESULTS: No significant differences were found in the genotype and allele frequencies of examined SOD2 and MTHFR gene variants between the elderly group and young controls (P > 0.05), nor when each gender was considered separately (P > 0.05). CONCLUSION: SOD2-9T/C and MTHFR 677C/T gene polymorphisms do not seem to be important in Jordanian population for longevity phenotype. PMID- 20003470 TI - Crystallographic structure of ubiquitin in complex with cadmium ions. AB - BACKGROUND: Ubiquitination plays a critical role in regulating many cellular processes, from DNA repair and gene transcription to cell cycle and apoptosis. It is catalyzed by a specific enzymatic cascade ultimately leading to the conjugation of ubiquitin to lysine residues of the target protein that can be the ubiquitin molecule itself and to the formation of poly-ubiquitin chains. FINDINGS: We present the crystal structure at 3.0 A resolution of bovine ubiquitin crystallized in presence of cadmium ions. Two molecules of ubiquitin are present in the asymmetric unit. Interestingly this non-covalent dimeric arrangement brings Lys-6 and Lys-63 of each crystallographically-independent monomer in close contact with the C-terminal ends of the other monomer. Residues Leu-8, Ile-44 and Val-70 that form a hydrophobic patch at the surface of the Ub monomer are trapped at the dimer interface. CONCLUSIONS: The structural basis for signalling by poly-Ub chains relies on a visualization of conformations of alternatively linked poly-Ub chains. This arrangement of ubiquitin could illustrate how linkages involving Lys-6 or Lys-63 of ubiquitin are produced in the cell. It also details how ubiquitin molecules can specifically chelate cadmium ions. PMID- 20003471 TI - Informed consent from patients participating in medical education: a survey from a university hospital in Jamaica. AB - BACKGROUND: Medical students at the University of the West Indies receive clinical training by passing through a series of hospital rotations at the University Hospital of the West Indies (UHWI). Many of these patients are unaware that medical students may be involved in their care. We performed this study to determine patient awareness and their willingness to participate in research and teaching activities. FINDINGS: All consecutive patients admitted to the UHWI between May 1, 2006 and May 29, 2006 who required elective or emergency surgical procedures were prospectively identified These patients were interviewed using a standardised pre-tested questionnaire about their knowledge and willingness to have medical students participate in the delivery of their hospital care. Data was analyzed using SPSS Version 12.0. There were 83 (39.5%) males and 127 (60.5%) females interviewed. The patients were unaware of the grade of the medical professional performing their interview/examination at admission in 157 (74.8%) cases or the grade of medical professional performing their operations in 101 (48.1%) cases. Only 14 (6.7%) patients were specifically asked to allow medical students to be present during their clinical evaluation and care. When specifically asked, 1 patient declined. Had they been asked, 196 (93.3%) patients would have voluntarily allowed medical student involvement. Only 90 (42.9%) patients were made aware that they were admitted to an academic centre with research interests. Only 6 (6.7%) patients declined. Had they been asked, 84 (93.3%) patients would be willing to participate in teaching or research projects. CONCLUSIONS: As medical educators, we are responsible to adhere to ethical and legal guidelines when we interact with patients. It is apparent that there is urgent need for policy development at the UWI to guide clinicians and students on their interactions with patients. PMID- 20003472 TI - Early severe morbidity and resource utilization in South African adults on antiretroviral therapy. AB - BACKGROUND: High rates of mortality and morbidity have been described in sub Saharan African patients within the first few months of starting highly active antiretroviral therapy (HAART). There is limited data on the causes of early morbidity on HAART and the associated resource utilization. METHODS: A cross sectional study was conducted of medical admissions at a secondary-level hospital in Cape Town, South Africa. Patients on HAART were identified from a register and HIV-infected patients not on HAART were matched by gender, month of admission, and age group to correspond with the first admission of each case. Primary reasons for admission were determined by chart review. Direct health care costs were determined from the provider's perspective. RESULTS: There were 53 in the HAART group with 70 admissions and 53 in the no-HAART group with 60 admissions. The median duration of HAART was 1 month (interquartile range 1-3 months). Median baseline CD4 count in the HAART group was 57 x 106 cells/L (IQR 15-115). The primary reasons for admission in the HAART group were more likely to be due to adverse drug reactions and less likely to be due to AIDS events than the no-HAART group (34% versus 7%; p < 0.001 and 39% versus 63%; p = 0.005 respectively). Immune reconstitution inflammatory syndrome was the primary reason for admission in 10% of the HAART group. Lengths of hospital stay per admission and inpatient survival were not significantly different between the two groups. Five of the 15 deaths in the HAART group were due to IRIS or adverse drug reactions. Median costs per admission of diagnostic and therapeutic services (laboratory investigations, radiology, intravenous fluids and blood, and non-ART medications) were higher in the HAART group compared with the no-HAART group (US$190 versus US$111; p = 0.001), but the more expensive non-curative costs (overhead, capital, and clinical staff) were not significantly different (US$1199 versus US$1128; p = 0.525). CONCLUSIONS: Causes of early morbidity are different and more complex in HIV-infected patients on HAART. This results in greater resource utilization of diagnostic and therapeutic services. PMID- 20003473 TI - Variable expression of cysteinyl leukotriene type I receptor splice variants in asthmatic females with different promoter haplotypes. AB - BACKGROUND: Cysteinyl leukotrienes are potent inflammatory mediators implicated in the pathogenesis of asthma. Human cysteinyl leukotriene receptor 1 (CYSLTR1) gene contains five exons that are variably spliced. Within its promoter few polymorphisms were described. To date, there has been no evidence about the expression of different splice variants of CysLT1 in asthma and their association with CYSLTR1 promoter polymorphisms.The goal of our study was to investigate CysLT1 alternative transcripts expression in asthmatic patients with different CYSLTR1 promoter haplotypes.The study groups consisted of 44 patients with asthma, diagnosed according to GINA 2008 criteria and 18 healthy subjects. Genomic DNA and total RNA was extracted from peripheral blood mononuclear cells. Real-time PCR was performed with specific primers for transcript I [GenBank:DQ131799] and II [GenBank:DQ131800]. Fragments of the CYSLTR1 promoter were amplified by PCR and sequenced directly to identify four single nucleotide polymorphisms: C/T [SNP:rs321029], A/C [SNP:rs2637204], A/G [SNP:rs2806489] and C/T [SNP:rs7066737]. RESULTS: The expression of CysLT1 transcript I and II in asthma did not differ from its expression in healthy control group. However, in major alleles homozygotic CAAC/CAAC women with asthma we found significantly higher expression of transcript I as compared to heterozygous CAAC/TCGC women in that loci. CysLT1 transcript I expression tended to negative correlation with episodes of acute respiratory infection in our asthmatic population. Moreover, expression of CysLT1 transcript II in CAAC/CAAC homozygotic women with asthma was significantly lower than in CAAC/CAAC healthy control females. CONCLUSIONS: Genetic variants of CYSLTR1 promoter might be associated with gender specific expression of CysLT1 alternative transcripts in patients with asthma. CysLT1 splice variants expression might also correlate with the susceptibility to infection in asthmatic population. PMID- 20003474 TI - Demographic characteristics of patients using a fully integrated psychosocial support service for cancer patients. AB - BACKGROUND: Psychosocial support services are an important component of modern cancer treatment. A major challenge for all psychosocial services is the achievement of equity of use. Previous studies in the UK have found that women of higher socio-economic status with breast cancer were over-represented amongst those accessing support services. People with other cancer diagnoses, those from socio-economically deprived areas, and men, were under-represented. FINDINGS: The Oncology Health Service, Kingston Upon Hull, UK, delivers fully integrated psychosocial support and interventions. To assess equity of access in this service, a cross-sectional study of all patients with cancer accessing the service during a 5 day period was carried out. One hundred and forty-five patients attended. Forty four percent were male, and the types of cancer were broadly in the proportions expected on the basis of population prevalence (breast cancer 22%, colorectal cancer 21%, lung cancer 16%). Sixty six percent came from the three most deprived quintiles of the Townsend deprivation Index. CONCLUSIONS: The fully integrated Oncology Health Service in Hull is accessed by a more diverse range of patients than previously reported for other services, and is an example of a model of service by which socially equitable use of psychosocial support in the National Health Service might be achieved. PMID- 20003475 TI - Unraveling the genomic diversity of small eukaryotes. AB - A report of the meeting Comparative Genomics of Eukaryotic Microorganisms, 17-22 October 2009, San Feliu de Guixols, Spain. PMID- 20003476 TI - Focus on collagen: in vitro systems to study fibrogenesis and antifibrosis state of the art. AB - Fibrosis represents a major global disease burden, yet a potent antifibrotic compound is still not in sight. Part of the explanation for this situation is the difficulties that both academic laboratories and research and development departments in the pharmaceutical industry have been facing in re-enacting the fibrotic process in vitro for screening procedures prior to animal testing. Effective in vitro characterization of antifibrotic compounds has been hampered by cell culture settings that are lacking crucial cofactors or are not holistic representations of the biosynthetic and depositional pathway leading to the formation of an insoluble pericellular collagen matrix. In order to appreciate the task which in vitro screening of antifibrotics is up against, we will first review the fibrotic process by categorizing it into events that are upstream of collagen biosynthesis and the actual biosynthetic and depositional cascade of collagen I. We point out oversights such as the omission of vitamin C, a vital cofactor for the production of stable procollagen molecules, as well as the little known in vitro tardy procollagen processing by collagen C-proteinase/BMP 1, another reason for minimal collagen deposition in cell culture. We review current methods of cell culture and collagen quantitation vis-a-vis the high content options and requirements for normalization against cell number for meaningful data retrieval. Only when collagen has formed a fibrillar matrix that becomes cross-linked, invested with ligands, and can be remodelled and resorbed, the complete picture of fibrogenesis can be reflected in vitro. We show here how this can be achieved. A well thought-out in vitro fibrogenesis system represents the missing link between brute force chemical library screens and rational animal experimentation, thus providing both cost-effectiveness and streamlined procedures towards the development of better antifibrotic drugs. PMID- 20003477 TI - Evidence of the adaptive evolution of immune genes in chicken. AB - The basis for understanding the characteristics of gene functional categories in chicken has been enhanced by the ongoing sequencing of the zebra finch genome, the second bird species to be extensively sequenced. This sequence provides an avian context for examining how variation in chicken has evolved since its divergence from its common ancestor with zebra finch as well as well as a calibrating point for studying intraspecific diversity within chicken. Immune genes have been subject to many selective processes during their evolutionary history: this gene class was investigated here in a set of orthologous chicken and zebra finch genes with functions assigned from the human ortholog. Tests demonstrated that nonsynonymous sites at immune genes were highly conserved both in chicken and on the avian lineage. McDonald-Kreitman tests provided evidence of adaptive evolution and a higher rate of selection on fixation of nonsynonymous substitutions at immune genes compared to that at non-immune genes. Further analyses showed that GC content was much higher in chicken than in zebra finch genes, and was significantly elevated in both species' immune genes. Pathogen challenges are likely to have driven the selective forces that have shaped variation at chicken immune genes, and continue to restrict diversity in this functional class. PMID- 20003478 TI - Overweight and obesity in urban Africa: A problem of the rich or the poor? AB - BACKGROUND: Obesity is a well recognized risk factor for various chronic diseases such as cardiovascular diseases, hypertension, and type 2 diabetes mellitus. The aim of this study was to shed light on the patterns of overweight and obesity in sub-Saharan Africa, with special interest in differences between the urban poor and the urban non-poor. The specific goals were to describe trends in overweight and obesity among urban women; and examine how these trends vary by education and household wealth. METHODS: The paper used Demographic and Health Surveys data from seven African countries where two surveys had been carried out with an interval of at least 10 years between them. Among the countries studied, the earliest survey took place in 1992 and the latest in 2005. The dependent variable was body mass index coded as: Not overweight/obese; Overweight; Obese. The key covariates were time lapse between the two surveys; woman's education; and household wealth. Control variables included working status, age, marital status, parity, and country. Multivariate ordered logistic regression in the context of the partial proportional odds model was used. RESULTS: Descriptive results showed that the prevalence of urban overweight/obesity increased by nearly 35% during the period covered. The increase was higher among the poorest (+50%) than among the richest (+7%). Importantly, there was an increase of 45-50% among the non educated and primary-educated women, compared to a drop of 10% among women with secondary education or higher. In the multivariate analysis, the odds ratio of the variable time lapse was 1.05 (p < 0.01), indicating that the prevalence of overweight/obesity increased by about 5% per year on average in the countries in the study. While the rate of change in urban overweight/obesity did not significantly differ between the poor and the rich, it was substantially higher among the non-educated women than among their educated counterparts. CONCLUSION: Overweight and obesity are on the rise in Africa and might take epidemic proportions in the near future. Like several other public health challenges, overweight and obesity should be tackled and prevented early as envisioned in the WHO Global strategy on diet, physical activity and health. PMID- 20003479 TI - In-vitro mapping of E-fields induced near pacemaker leads by simulated MR gradient fields. AB - BACKGROUND: Magnetic resonance imaging (MRI) of patients with implanted cardiac pacemakers is generally contraindicated but some clinicians condone scanning certain patients. We assessed the risk of inducing unintended cardiac stimulation by measuring electric fields (E) induced near lead tips by a simulated MRI gradient system. The objectives of this study are to map magnetically induced E near distal tips of leads in a saline tank to determine the spatial distribution and magnitude of E and compare them with E induced by a pacemaker pulse generator (PG). METHODS: We mapped magnetically induced E with 0.1 mm resolution as close as 1 mm from lead tips. We used probes with two straight electrodes (e.g. wire diameter of 0.2 mm separated by 0.9 mm). We generated magnetic flux density (B) with a Helmholtz coil throughout 0.6% saline in a 24 cm diameter tank with (dB/dt) of 1 T/sec (1 kHz sinusoidal waveform). Separately, we measured E near the tip of leads when connected to a PG set to a unipolar mode. Measurements were non-invasive (not altering the leads or PG under study). RESULTS: When scaled to 30 T/s (a clinically relevant value), magnetically-induced E exceeded the E produced by a PG. The magnetically-induced E only occurred when B was coincident with or within 15 msec of implantable pacemaker's pulse. CONCLUSIONS: Potentially hazardous situations are possible during an MR scan due to gradient fields. Unintended stimulation can be induced via abandoned leads and leads connected to a pulse generator with loss of hermetic seal at the connector. Also, pacemaker dependent patients can receive drastically altered pacing pulses. PMID- 20003480 TI - Free abdominal fluid without obvious solid organ injury upon CT imaging: an actual problem or simply over-diagnosing? AB - Whereas a non-operative approach for hemodynamically stable patients with free intraabdominal fluid in the presence of solid organ injury is generally accepted, the presence of free fluid in the abdomen without evidence of solid organ injury not only presents a challenge for the treating emergency physician but also for the surgeon in charge. Despite recent advances in imaging modalities, with multi detector computed tomography (CT) (with or without contrast agent) usually the imaging method of choice, diagnosis and interpretation of the results remains difficult. While some studies conclude that CT is highly accurate and relatively specific at diagnosing mesenteric and hollow viscus injury, others studies deem CT to be unreliable. These differences may in part be due to the experience and the interpretation of the radiologist and/or the treating physician or surgeon.A search of the literature has made it apparent that there is no straightforward answer to the question what to do with patients with free intraabdominal fluid on CT scanning but without signs of solid organ injury. In hemodynamically unstable patients, free intraabdominal fluid in the absence of solid organ injury usually mandates immediate surgical intervention. For patients with blunt abdominal trauma and more than just a trace of free intraabdominal fluid or for patients with signs of peritonitis, the threshold for a surgical exploration - preferably by a laparoscopic approach - should be low. Based on the available information, we aim to provide the reader with an overview of the current literature with specific emphasis on diagnostic and therapeutic approaches to this problem and suggest a possible algorithm, which might help with the adequate treatment of such patients. PMID- 20003481 TI - Defining the healthy "core microbiome" of oral microbial communities. AB - BACKGROUND: Most studies examining the commensal human oral microbiome are focused on disease or are limited in methodology. In order to diagnose and treat diseases at an early and reversible stage an in-depth definition of health is indispensible. The aim of this study therefore was to define the healthy oral microbiome using recent advances in sequencing technology (454 pyrosequencing). RESULTS: We sampled and sequenced microbiomes from several intraoral niches (dental surfaces, cheek, hard palate, tongue and saliva) in three healthy individuals. Within an individual oral cavity, we found over 3600 unique sequences, over 500 different OTUs or "species-level" phylotypes (sequences that clustered at 3% genetic difference) and 88 - 104 higher taxa (genus or more inclusive taxon). The predominant taxa belonged to Firmicutes (genus Streptococcus, family Veillonellaceae, genus Granulicatella), Proteobacteria (genus Neisseria, Haemophilus), Actinobacteria (genus Corynebacterium, Rothia, Actinomyces), Bacteroidetes (genus Prevotella, Capnocytophaga, Porphyromonas) and Fusobacteria (genus Fusobacterium).Each individual sample harboured on average 266 "species-level" phylotypes (SD 67; range 123 - 326) with cheek samples being the least diverse and the dental samples from approximal surfaces showing the highest diversity. Principal component analysis discriminated the profiles of the samples originating from shedding surfaces (mucosa of tongue, cheek and palate) from the samples that were obtained from solid surfaces (teeth).There was a large overlap in the higher taxa, "species-level" phylotypes and unique sequences among the three microbiomes: 84% of the higher taxa, 75% of the OTUs and 65% of the unique sequences were present in at least two of the three microbiomes. The three individuals shared 1660 of 6315 unique sequences. These 1660 sequences (the "core microbiome") contributed 66% of the reads. The overlapping OTUs contributed to 94% of the reads, while nearly all reads (99.8%) belonged to the shared higher taxa. CONCLUSIONS: We obtained the first insight into the diversity and uniqueness of individual oral microbiomes at a resolution of next-generation sequencing. We showed that a major proportion of bacterial sequences of unrelated healthy individuals is identical, supporting the concept of a core microbiome at health. PMID- 20003482 TI - Manual annotation and analysis of the defensin gene cluster in the C57BL/6J mouse reference genome. AB - BACKGROUND: Host defense peptides are a critical component of the innate immune system. Human alpha- and beta-defensin genes are subject to copy number variation (CNV) and historically the organization of mouse alpha-defensin genes has been poorly defined. Here we present the first full manual genomic annotation of the mouse defensin region on Chromosome 8 of the reference strain C57BL/6J, and the analysis of the orthologous regions of the human and rat genomes. Problems were identified with the reference assemblies of all three genomes. Defensins have been studied for over two decades and their naming has become a critical issue due to incorrect identification of defensin genes derived from different mouse strains and the duplicated nature of this region. RESULTS: The defensin gene cluster region on mouse Chromosome 8 A2 contains 98 gene loci: 53 are likely active defensin genes and 22 defensin pseudogenes. Several TATA box motifs were found for human and mouse defensin genes that likely impact gene expression. Three novel defensin genes belonging to the Cryptdin Related Sequences (CRS) family were identified. All additional mouse defensin loci on Chromosomes 1, 2 and 14 were annotated and unusual splice variants identified. Comparison of the mouse alpha-defensins in the three main mouse reference gene sets Ensembl, Mouse Genome Informatics (MGI), and NCBI RefSeq reveals significant inconsistencies in annotation and nomenclature. We are collaborating with the Mouse Genome Nomenclature Committee (MGNC) to establish a standardized naming scheme for alpha defensins. CONCLUSIONS: Prior to this analysis, there was no reliable reference gene set available for the mouse strain C57BL/6J defensin genes, demonstrating that manual intervention is still critical for the annotation of complex gene families and heavily duplicated regions. Accurate gene annotation is facilitated by the annotation of pseudogenes and regulatory elements. Manually curated gene models will be incorporated into the Ensembl and Consensus Coding Sequence (CCDS) reference sets. Elucidation of the genomic structure of this complex gene cluster on the mouse reference sequence, and adoption of a clear and unambiguous naming scheme, will provide a valuable tool to support studies on the evolution, regulatory mechanisms and biological functions of defensins in vivo. PMID- 20003483 TI - Motor ability in children treated for idiopathic clubfoot. A controlled pilot study. AB - BACKGROUND: To study motor ability at seven years of age in children treated for idiopathic clubfoot and its relation to clubfoot laterality, foot status and the amount of surgery performed. METHODS: Twenty children (mean age 7.5 years, SD 3.2 months) from a consecutive birth cohort from our hospital catchments area (300.000 inhabitants from southern Sweden) were assessed with the Movement Assessment Battery for Children (MABC) and the Clubfoot Assessment Protocol (CAP). RESULTS: Compared to typically developing children an increased prevalence of motor impairment was found regarding both the total score for MABC (p < 0.05) and the subtest ABC-Ball skills (p < 0.05). No relationship was found between the child's actual foot status, laterality or the extent of foot surgery with the motor ability as measured with MABC. Only the CAP item "one-leg stand" correlated significantly with the MABC (rs = -0.53, p = 0.02). CONCLUSIONS: Children with idiopathic clubfoot appear to have an increased risk of motor activity limitations and it is possible that other factors, independent of the clinical status, might be involved. The ability to keep balance on one leg may be a sufficient tool for determining which children in the orthopedic setting should be more thoroughly evaluated regarding their neuromotor functioning. PMID- 20003484 TI - Penicillin-binding proteins (PBP) and Lmo0441 (a PBP-like protein) play a role in Beta-lactam sensitivity of Listeria monocytogenes. AB - While seven penicillin-binding proteins (PBPs) or PBP-like proteins have been identified either by radiolabelled penicillin binding studies or genomic analysis, only PBP3 has been considered of interest for Beta-lactams activity against Listeria monocytogenes. Herein we reveal that both PBP4 and Lmo0441 (a PBP-like protein) play a direct role in cephalosporin activity in L. monocytogenes while PBP4 additionally has a protective affect against both penicillin and carbapenem. PMID- 20003487 TI - Recent developments in podiatric prescribing in the UK and Australia. AB - Recent and substantial changes in access to restricted medicines by podiatrists in Australia are clearly consistent with healthcare policies aimed at reshaping the healthcare workforce. At the same time, prescribing and access to medicines by allied health professionals, including podiatrists, has been the focus of a recent scoping project by the UK Department of Health. In this commentary we explore the possible implications of these changes. PMID- 20003486 TI - Phlegmonous colitis: another source of sepsis in cirrhotic patients? AB - BACKGROUND: The clinical relevance of phlegmonous colitis (PC), a rare autopsy finding in cirrhotic patients, is poorly documented. We postulated that PC might be a source of sepsis in patients with portal hypertensive colopathy (PHC). CASE PRESENTATION: We report three cirrhotic patients who were admitted with abdominal sepsis and who illustrate, to various degrees, the clinico-pathological sequence of colonic alterations associated with portal hypertension. Two cirrhotic patients with PHC developed gram-negative bacteraemia and quickly responded to intravenous antibiotics. Another cirrhotic patient underwent emergency colectomy for PC, and subsequently died from multiple organ failure. Histological alterations in the operative specimen included: a) mucosal ulcerations; b) disseminated micro-abscesses in the submucosa; and c) a severe vasculopathy leading to complete obliteration of submucosal blood vessels. CONCLUSIONS: These data suggest that cirrhotic patients with PHC may progress towards PC, which, in turn, may be the cause for life-threatening sepsis. PMID- 20003485 TI - DNA double strand break repair enzymes function at multiple steps in retroviral infection. AB - BACKGROUND: DNA double strand break (DSB) repair enzymes are thought to be necessary for retroviral infection, especially for the post-integration repair and circularization of viral cDNA. However, the detailed roles of DSB repair enzymes in retroviral infection remain to be elucidated. RESULTS: A GFP reporter assay showed that the infectivity of an HIV-based vector decreased in ATM- and DNA-PKcs-deficient cells when compared with their complemented cells, while that of an MLV-based vector was diminished in Mre11- and DNA-PKcs-deficient cells. By using a method based on inverse- and Alu-PCR, we analyzed sequences around 3' HIV 1 integration sites in ATM-, Mre11- and NBS1- deficient cells. Increased abnormal junctions between the HIV-1 provirus and the host DNA were found in these mutant cell lines compared to the complemented cell lines and control MRC5SV cells. The abnormal junctions contained two types of insertions: 1) GT dinucleotides, which are normally removed by integrase during integration, and 2) inserted nucleotides of unknown origin. Artemis-deficient cells also showed such abnormalities. In Mre11-deficient cells, part of a primer binding site sequence was also detected. The 5' host-virus junctions in the mutant cells also contained these types of abnormal nucleotides. Moreover, the host-virus junctions of the MLV provirus showed similar abnormalities. These findings suggest that DSB repair enzymes play roles in the 3'-processing reaction and protection of the ends of viral DNA after reverse transcription. We also identified both 5' and 3' junctional sequences of the same provirus by inverse PCR and found that only the 3' junctions were abnormal with aberrant short repeats, indicating that the integration step was partially impaired in these cells. Furthermore, the conserved base preferences around HIV-1 integration sites were partially altered in ATM-deficient cells. CONCLUSIONS: These results suggest that DSB repair enzymes are involved in multiple steps including integration and pre-integration steps during retroviral replication. PMID- 20003488 TI - Trends and variation in the management of oesophagogastric cancer patients: a population-based survey. AB - BACKGROUND: Previous evidence indicates potential variation in the quality of care of cancer patients. We aimed to examine whether recent changes in the treatment of oesophagogastric cancers have been distributed equally among different patient subgroups. METHODS: We analysed population-based cancer registry data about the treatment patterning of oesophagogastric cancer (other than oesophageal squamous cell carcinoma) during 1995-2006. RESULTS: There were 14,077 patients aged > or =40 years (69% men). There was only limited information on stage, and no information on co-morbidity status. During successive triennia, curative surgery use decreased from 28% to 20% (p < 0.001) whilst chemotherapy use increased from 9% to 30% (p < 0.001). Use of palliative surgery and of radiotherapy increased significantly but modestly (7% to 10%, and 9% to 11%, respectively). In multivariable logistic regression adjusting for age group, gender, diagnosis period and tumour type, curative surgery and chemotherapy were used less frequently in more deprived patients [per increasing deprivation group Odds Ratio (OR) = 0.96, 95% Confidence Interval (CI) 0.93-0.99, and OR = 0.90, 95%CI 0.87-0.93, respectively, p < 0.001 for both)]. Chemotherapy was also used less frequently in women (OR = 0.76, p < 0.001). CONCLUSIONS: During the study period, curative surgery decreased by a third and chemotherapy use increased by more than three-fold, reflecting improvements in the appropriateness and quality of management, but chemotherapy use, in particular, was unequal, both by socioeconomic status and gender. PMID- 20003489 TI - Severity of Giardia infection associated with post-infectious fatigue and abdominal symptoms two years after. AB - BACKGROUND: A high rate of post-infectious fatigue and abdominal symptoms two years after a waterborne outbreak of giardiasis in Bergen, Norway in 2004 has previously been reported. The aim of this report was to identify risk factors associated with such manifestations. METHODS: All laboratory confirmed cases of giardiasis (n = 1262) during the outbreak in Bergen in 2004 received a postal questionnaire two years after. Degree of post-infectious abdominal symptoms and fatigue, as well as previous abdominal problems, was recorded. In the statistical analyses number of treatment courses, treatment refractory infection, delayed education and sick leave were used as indices of protracted and severe Giardia infection. Age, gender, previous abdominal problems and symptoms during infection were also analysed as possible risk factors. Simple and multiple ordinal logistic regression models were used for the analyses. RESULTS: The response rate was 81% (1017/1262), 64% were women and median age was 31 years (range 3-93), compared to 61% women and 30 years (range 2-93) among all 1262 cases. Factors in multiple regression analysis significantly associated with abdominal symptoms two years after infection were: More than one treatment course, treatment refractory infection, delayed education, bloating and female gender. Abdominal problems prior to Giardia infection were not associated with post-infectious abdominal symptoms. More than one treatment course, delayed education, sick leave more than 2 weeks, and malaise at the time of infection, were significantly associated with fatigue in the multiple regression analysis, as were increasing age and previous abdominal problems. CONCLUSION: Protracted and severe giardiasis seemed to be a risk factor for post-infectious fatigue and abdominal symptoms two years after clearing the Giardia infection. PMID- 20003491 TI - The Retrovirology Open Access experience. AB - The Retrovirology Open Access experience after publishing more than 500 articles is discussed. PMID- 20003490 TI - Sensitive HPV detection in oropharyngeal cancers. AB - BACKGROUND: Human papillomaviruses (HPV) are the aetiological agents of certain benign and malignant tumours of skin and mucosae; the most important of which is cervical cancer. Also, the incidence of ano-genital warts, HPV-anal cancer and oropharyngeal cancers are rising. To help ascertain a useful PCR detection protocol for oropharyngeal cancers, we directly compared three commonly used primer sets in detection of HPV from different clinical samples. METHODS: We compared PGMY09/11, MY09/11 and GP5+/6+ primers sets in PCRs of 34 clinically diagnosed samples of genital warts, cervical brushings (with associated histological diagnosis) and vulval biopsies. All negative samples were subsequently tested using the previously reported PGMY/GP PCR method and amplicons directly sequenced for confirmation and typing. An optimised PCR protocol was then compared to a line blot assay for detection of HPV in 15 oropharyngeal cancer samples. RESULTS: PGMY09/11 primers detected HPV presence in more cervical brushing (100%) and genital wart (92.9%) samples compared to MY09/11 (90% and 64.3%) and GP5+/6+ (80% and 64.3%) primer sets, respectively. From vulval biopsies, HPV detection rates were: MY09/11 (63.6%), GP5+/6+ (54.5%) and PGMY09/11 (54.5%). PGMY/GP nested PCR demonstrated that HPV was present, and direct sequencing confirmed genotypes. This nested PCR protocol showed detection of HPV in 10/15 (66.7%) of oropharyngeal cancer samples. CONCLUSIONS: PGMY09/11 primers are the preferred primer set among these three for primary PCR screening with different clinical samples. MY09/11 and GP5+/6+ may be used (particularly for cervical samples) but demonstrate lower detection rates. A nested PCR approach (i.e. a PGMY-GP system) may be required to confirm negativity or to detect low levels of HPV, undetectable using current primary PCR methods, as demonstrated using oropharyngeal cancer samples. PMID- 20003492 TI - A cross-curricular physical activity intervention to combat cardiovascular disease risk factors in 11-14 year olds: 'activity knowledge circuit'. AB - BACKGROUND: Cardiovascular disease is the leading cause of mortality worldwide. Risk factors associated with cardiovascular disease have been shown to track from childhood through to adulthood. Previous school-based physical activity interventions have demonstrated modest improvements to cardiovascular disease risk factors by implementing extra-curricular activities or improving current physical education curriculum. Few have attempted to increase physical activity in class-room taught curriculum subjects. This study will outline a school-based cross-curricular physical activity intervention to combat cardiovascular disease risk factors in 11-14 year old children. METHOD/DESIGN: A South Wales Valley school of low socio-economic status has been selected to take part. Participants from year eight (12-13 years) are to be assigned to an intervention group, with maturation-matched participants from years seven (11-12 years) and nine (13-14 years) assigned to a control group. A cross-curricular physical activity intervention will be implemented to increase activity by two hours a week for 18 weeks. Participants will briskly walk 3200 m twice weekly during curriculum lessons (60 minutes duration). With the exception of physical education, all curriculum subjects will participate, with each subject delivering four intervention lessons. The intervention will be performed outdoors and on school premises. An indoor course of equal distance will be used during adverse weather conditions. Cardiovascular disease risk factors will be measured pre- and post intervention for intervention and control groups. These will take place during physical education lessons and will include measures of stature, mass, waist, hip, and neck circumferences, together with skinfold measure's taken at four sites. Blood pressure will be measured, and fitness status assessed via the 20 m multi-stage fitness test. Questionnaires will be used to determine activity behaviour (physical activity questionnaire for adolescence), diet (seven day food diary) and maturation status. Fasting blood variables will include total cholesterol, low-density lipoprotein cholesterol, high density lipoprotein cholesterol, triglycerides, insulin, glucose, high-sensitivity C-reactive protein, interleukin-6, adiponectin, and fibrinogen. Motivational variables and psychological well-being will be assessed by questionnaire. DISCUSSION: Our study may prove to be a cost effective strategy to increase school time physical activity to combat cardiovascular disease risk factors in children. TRIAL REGISTRATION: [NCT00998478]. PMID- 20003493 TI - Using theatre in education in a traditional lecture oriented medical curriculum. AB - BACKGROUND: Lectures supported by theatrical performance may enhance learning and be an attractive alternative to traditional lectures. This study describes our experience with using theatre in education for medical students since 2001. METHODS: The volunteer students, coached by experienced students, were given a two-week preparation period to write and prepare different dramatized headache scenarios during three supervised meetings. A theatrical performance was followed by a student presentation about history taking and clinical findings in diagnosing headache. Finally, a group discussion led by students dealt with issues raised in the performance. The evaluation of the theatre in education lecture "A Primary Care Approach to Headache" was based on feedback from students. RESULTS: More than 90% of 43 responding students fully agreed with the statement "Theatrical performance made it easier to understand the topic". More than 90% disagreed with the statements "Lecture halls were not appropriate for this kind of interaction" and "Students as teachers were not appropriate". Open ended questions showed that the lesson was thought of as fun, good and useful by most students. The headache questions in the final exam showed results that were similar to average exam results for other questions. CONCLUSION: Using theatrical performance in medical education was appreciated by most students and may facilitate learning and enhance empathy and team work communication skills. PMID- 20003494 TI - PALB2 variants in hereditary and unselected Finnish prostate cancer cases. AB - BACKGROUND: PALB2 1592delT mutation is associated with increased breast cancer and suggestive prostate cancer (PRCA) risk in Finland. In this study we wanted to assess if any other PALB2 variants associate to increased PRCA risk and clinically describe patients with formerly found PALB2 1592delT mutation. METHODS: Finnish families with two or more PRCA cases (n = 178) and unselected cases (n = 285) with complete clinical data were initially screened for variants in the coding region and splice sites of PALB2. Potentially interesting variants were verified in additional set of unselected cases (n = 463). RESULTS: From our clinically defined sample set we identified total of six variants in PALB2. No novel variants among Finnish PRCA cases were found. Clinical characteristics of the variant carriers, including the previously described family carrying PALB2 1592delT, revealed a trend towards aggressive disease, which also applied to a few non-familial cases. Hypersensitivity to mitomycin C (MMC) of lymphoblasts from individuals from the family with 1592delT revealed haploinsufficiency among carriers with altered genotype. CONCLUSIONS: Though any of the detected PALB2 variants do not associate to PRCA in population level in Finland it cannot be ruled out that some of these variants contribute to cancer susceptibility at individual level. PMID- 20003495 TI - A novel mutation causing mild, atypical fumarylacetoacetase deficiency (Tyrosinemia type I): a case report. AB - A male patient, born to unrelated Belgian parents, presented at 4 months with epistaxis, haematemesis and haematochezia. On physical examination he presented petechiae and haematomas, and a slightly enlarged liver. Serum transaminases were elevated to 5-10 times upper limit of normal, alkaline phosphatases were 1685 U/L (<720), total bilirubin was 2.53 mg/dl (<1.0), ammonaemia 69 microM (<32), prothrombin time less than 10%, thromboplastin time >180 s (<60) and alpha fetoprotein 29723 microg/L (<186). Plasma tyrosine (651 microM) and methionine (1032 microM) were strongly increased. In urine, tyrosine metabolites and 4-oxo-6 hydroxyheptanoic acid were increased, but succinylacetone and succinylacetoacetate--pathognomonic for tyrosinemia type I--were repeatedly undetectable. Delta-aminolevulinic acid was normal, which is consistent with the absence of succinylacetone. Abdominal ultrasound and brain CT were normal.Fumarylacetoacetase (FAH) protein and activity in cultured fibroblasts and liver tissue were decreased but not absent. 4-hydroxyphenylpyruvate dioxygenase activity in liver was normal, which is atypical for tyrosinemia type I. A novel mutation was found in the FAH gene: c.103G>A (Ala35Thr). In vitro expression studies showed this mutation results in a strongly decreased FAH protein expression.Dietary treatment with phenylalanine and tyrosine restriction was initiated at 4 months, leading to complete clinical and biochemical normalisation. The patient, currently aged 12 years, shows a normal physical and psychomotor development.This is the first report of mild tyrosinemia type I disease caused by an Ala35Thr mutation in the FAH gene, presenting atypically without increase of the diagnostically important toxic metabolites succinylacetone and succinylacetoacetate. PMID- 20003496 TI - Traumatic fracture-dislocation of the hip following rugby tackle: a case report. AB - Posterior fracture-dislocation of hip is uncommonly encountered in rugby injuries. We report such a case in an adult while playing rugby. The treating orthopaedician can be caught unaware and injuries in such sports can be potentially misdiagnosed as hip sprains. Immediate reduction of the dislocation was performed in theatres. The fracture was fixed with two lag screws and a neutralization plate. This led to early rehabilitation and speedy recovery with return to sporting activities by 12 months. PMID- 20003497 TI - Blunt traumatic pericardial rupture and cardiac herniation with a penetrating twist: two case reports. AB - BACKGROUND: Blunt Traumatic Pericardial Rupture (BTPR) with resulting cardiac herniation following chest trauma is an unusual and often fatal condition. Although there has been a multitude of case reports of this condition in past literature, the recurring theme is that of a missed injury. Its occurrence in severe blunt trauma is in the order of 0.4%. It is an injury that frequently results in pre/early hospital death and diagnosis at autopsy, probably owing to a combination of diagnostic difficulties, lack of familiarity and associated polytrauma. Of the patients who survive to hospital attendance, the mortality rate is in the order of 57-64%. METHODS: We present two survivors of BTPR and cardiac herniation, one with a delayed penetrating cardiac injury secondary to rib fractures. With these two cases and literature review, we hope to provide a greater awareness of this injury CONCLUSION: BTPR and cardiac herniation is a complex and often fatal injury that usually presents under the umbrella of polytrauma. Clinicians must maintain a high index of suspicion for BTPR but, even then, the diagnosis is fraught with difficulty. In blunt chest trauma, patients should be considered high risk for BTPR when presenting with:Cardiovascular instability with no obvious cause. Prominent or displaced cardiac silhouette and asymmetrical large volume pneumopericardium. Potentially, with increasing awareness of the injury and improved use and availability of imaging modalities, the survival rates will improve and cardiac Herniation could even be considered the 5th H of reversible causes of blunt traumatic PEA arrest. PMID- 20003498 TI - Separating therapeutic efficacy from glucocorticoid side-effects in rodent arthritis using novel, liposomal delivery of dexamethasone phosphate: long-term suppression of arthritis facilitates interval treatment. AB - INTRODUCTION: Glucocorticoids have extensively been used in the treatment of rheumatoid arthritis and other inflammatory diseases. However, their side-effects remain the major limitation in clinical use and an improved therapeutic index is needed. METHODS: Therapeutic efficacy and persistence of free and liposomal dexamethasone phosphate (DXM-P) were determined in mouse collagen-induced arthritis. For regimens with equal therapeutic benefit, the side-effect profiles were analysed over time with respect to collagen breakdown, suppression of the hypothalamus-pituitary-adrenal (HPA) axis, changes in blood glucose levels and the haematological profile. In addition, the presence of drug was monitored in plasma. RESULTS: Liposomal DXM-P, but not free drug, resulted in a persistent anti-inflammatory effect. Comparable clinical benefit was achieved with a single administration of 4 mg/kg liposomal DXM-P or daily administrations of 1.6 mg/kg free drug for at least 7 days. For the liposomal form, but not for the free form, we observed a limitation of the suppression of the HPA axis in time and an absence of the drug-induced gluconeogenesis. CONCLUSIONS: Liposomal DXM-P, but not free DXM-P, achieves therapeutic persistence in mouse collagen-induced arthritis, which results in drug-free periods of therapeutic benefit. The physical absence of drug after day 2 is associated with a reduction of the typical glucocorticoid side-effects profile. Liposomal DXM-P thereby has an improved therapeutic window. PMID- 20003499 TI - Impaired verbal memory in Parkinson disease: relationship to prefrontal dysfunction and somatosensory discrimination. AB - OBJECTIVE: To study the neurocognitive profile and its relationship to prefrontal dysfunction in non-demented Parkinson's disease (PD) with deficient haptic perception. METHODS: Twelve right-handed patients with PD and 12 healthy control subjects underwent thorough neuropsychological testing including Rey complex figure, Rey auditory verbal and figural learning test, figural and verbal fluency, and Stroop test. Test scores reflecting significant differences between patients and healthy subjects were correlated with the individual expression coefficients of one principal component, obtained in a principal component analysis of an oxygen-15-labeled water PET study exploring somatosensory discrimination that differentiated between the two groups and involved prefrontal cortices. RESULTS: We found significantly decreased total scores for the verbal learning trials and verbal delayed free recall in PD patients compared with normal volunteers. Further analysis of these parameters using Spearman's ranking correlation showed a significantly negative correlation of deficient verbal recall with expression coefficients of the principal component whose image showed a subcortical-cortical network, including right dorsolateral-prefrontal cortex, in PD patients. CONCLUSION: PD patients with disrupted right dorsolateral prefrontal cortex function and associated diminished somatosensory discrimination are impaired also in verbal memory functions. A negative correlation between delayed verbal free recall and PET activation in a network including the prefrontal cortices suggests that verbal cues and accordingly declarative memory processes may be operative in PD during activities that demand sustained attention such as somatosensory discrimination. Verbal cues may be compensatory in nature and help to non-specifically enhance focused attention in the presence of a functionally disrupted prefrontal cortex. PMID- 20003500 TI - BLAST+: architecture and applications. AB - BACKGROUND: Sequence similarity searching is a very important bioinformatics task. While Basic Local Alignment Search Tool (BLAST) outperforms exact methods through its use of heuristics, the speed of the current BLAST software is suboptimal for very long queries or database sequences. There are also some shortcomings in the user-interface of the current command-line applications. RESULTS: We describe features and improvements of rewritten BLAST software and introduce new command-line applications. Long query sequences are broken into chunks for processing, in some cases leading to dramatically shorter run times. For long database sequences, it is possible to retrieve only the relevant parts of the sequence, reducing CPU time and memory usage for searches of short queries against databases of contigs or chromosomes. The program can now retrieve masking information for database sequences from the BLAST databases. A new modular software library can now access subject sequence data from arbitrary data sources. We introduce several new features, including strategy files that allow a user to save and reuse their favorite set of options. The strategy files can be uploaded to and downloaded from the NCBI BLAST web site. CONCLUSION: The new BLAST command-line applications, compared to the current BLAST tools, demonstrate substantial speed improvements for long queries as well as chromosome length database sequences. We have also improved the user interface of the command-line applications. PMID- 20003501 TI - Reversal of childhood idiopathic scoliosis in an adult, without surgery: a case report and literature review. AB - BACKGROUND: Some patients with mild or moderate thoracic scoliosis (Cobb angle <50-60 degrees) suffer disproportionate impairment of pulmonary function associated with deformities in the sagittal plane and reduced flexibility of the spine and chest cage. Long-term improvement in the clinical signs and symptoms of childhood onset scoliosis in an adult, without surgical intervention, has not been documented previously. CASE PRESENTATION: A diagnosis of thoracic scoliosis (Cobb angle 45 degrees) with pectus excavatum and thoracic hypokyphosis in a female patient (DOB 9/17/52) was made in June 1964. Immediate spinal fusion was strongly recommended, but the patient elected a daily home exercise program taught during a 6-week period of training by a physical therapist. This regime was carried out through 1992, with daily aerobic exercise added in 1974. The Cobb angle of the primary thoracic curvature remained unchanged. Ongoing clinical symptoms included dyspnea at rest and recurrent respiratory infections. A period of multimodal treatment with clinical monitoring and treatment by an osteopathic physician was initiated when the patient was 40 years old. This included deep tissue massage (1992-1996); outpatient psychological therapy (1992-1993); a daily home exercise program focused on mobilization of the chest wall (1992-2005); and manipulative medicine (1994-1995, 1999-2000). Progressive improvement in chest wall excursion, increased thoracic kyphosis, and resolution of long-standing respiratory symptoms occurred concomitant with a >10 degree decrease in Cobb angle magnitude of the primary thoracic curvature. CONCLUSION: This report documents improved chest wall function and resolution of respiratory symptoms in response to nonsurgical approaches in an adult female, diagnosed at age eleven years with idiopathic scoliosis. PMID- 20003502 TI - Ethnobotanical knowledge in rural communities of Cordoba (Argentina): the importance of cultural and biogeographical factors. AB - BACKGROUND: The possibility to better understand the relationships within the men, the nature and their culture has extreme importance because allows the characterisation of social systems through their particular environmental perception, and provides useful tools for the development of conservation policies. METHODS: The present study was planned to disentangle environmental and cultural factors that are influencing the perception, knowledge and uses of edible and medicinal plants in rural communities of Cordoba (Argentina). Interviews an participant observation were conducted in nine rural communities located in three different biogeographical areas. Data about knowledge of medicinal and edible plants and sociocultural variables were obtained. Data were analysed by Principal Components Analysis (PCA). RESULTS: The analysis of data confirmed that medicinal species are widely used whereas the knowledge on edible plants is eroding. The PCA showed four groups of communities, defined by several particular combinations of sociocultural and/or natural variables. CONCLUSION: This comprehensive approach suggests that in general terms the cultural environment has a stronger influence than the natural environment on the use of medicinal and edible plants in rural communities of Cordoba (Argentina). PMID- 20003503 TI - A transcriptomic computational analysis of mastic oil-treated Lewis lung carcinomas reveals molecular mechanisms targeting tumor cell growth and survival. AB - BACKGROUND: Mastic oil from Pistacia lentiscus variation chia, a blend of bioactive terpenes with recognized medicinal properties, has been recently shown to exert anti-tumor growth activity through inhibition of cancer cell proliferation, survival, angiogenesis and inflammatory response. However, no studies have addressed its mechanisms of action at genome-wide gene expression level. METHODS: To investigate molecular mechanisms triggered by mastic oil, Lewis Lung Carcinoma cells were treated with mastic oil or DMSO and RNA was collected at five distinct time points (3-48 h). Microarray expression profiling was performed using Illumina mouse-6 v1 beadchips, followed by computational analysis. For a number of selected genes, RT-PCR validation was performed in LLC cells as well as in three human cancer cell lines of different origin (A549, HCT116, K562). PTEN specific inhibition by a bisperovanadium compound was applied to validate its contribution to mastic oil-mediated anti-tumor growth effects. RESULTS: In this work we demonstrated that exposure of Lewis lung carcinomas to mastic oil caused a time-dependent alteration in the expression of 925 genes. GO analysis associated expression profiles with several biological processes and functions. Among them, modifications on cell cycle/proliferation, survival and NF kappaB cascade in conjunction with concomitant regulation of genes encoding for PTEN, E2F7, HMOX1 (up-regulation) and NOD1 (down-regulation) indicated some important mechanistic links underlying the anti-proliferative, pro-apoptotic and anti-inflammatory effects of mastic oil. The expression profiles of Hmox1, Pten and E2f7 genes were similarly altered by mastic oil in the majority of test cancer cell lines. Inhibition of PTEN partially reversed mastic oil effects on tumor cell growth, indicating a multi-target mechanism of action. Finally, k means clustering, organized the significant gene list in eight clusters demonstrating a similar expression profile. Promoter analysis in a representative cluster revealed shared putative cis-elements suggesting a common regulatory transcription mechanism. CONCLUSIONS: Present results provide novel evidence on the molecular basis of tumor growth inhibition mediated by mastic oil and set a rational basis for application of genomics and bioinformatic methodologies in the screening of natural compounds with potential cancer chemopreventive activities. PMID- 20003504 TI - Quantitative comparison of microarray experiments with published leukemia related gene expression signatures. AB - BACKGROUND: Multiple gene expression signatures derived from microarray experiments have been published in the field of leukemia research. A comparison of these signatures with results from new experiments is useful for verification as well as for interpretation of the results obtained. Currently, the percentage of overlapping genes is frequently used to compare published gene signatures against a signature derived from a new experiment. However, it has been shown that the percentage of overlapping genes is of limited use for comparing two experiments due to the variability of gene signatures caused by different array platforms or assay-specific influencing parameters. Here, we present a robust approach for a systematic and quantitative comparison of published gene expression signatures with an exemplary query dataset. RESULTS: A database storing 138 leukemia-related published gene signatures was designed. Each gene signature was manually annotated with terms according to a leukemia-specific taxonomy. Two analysis steps are implemented to compare a new microarray dataset with the results from previous experiments stored and curated in the database. First, the global test method is applied to assess gene signatures and to constitute a ranking among them. In a subsequent analysis step, the focus is shifted from single gene signatures to chromosomal aberrations or molecular mutations as modeled in the taxonomy. Potentially interesting disease characteristics are detected based on the ranking of gene signatures associated with these aberrations stored in the database. Two example analyses are presented. An implementation of the approach is freely available as web-based application. CONCLUSIONS: The presented approach helps researchers to systematically integrate the knowledge derived from numerous microarray experiments into the analysis of a new dataset. By means of example leukemia datasets we demonstrate that this approach detects related experiments as well as related molecular mutations and may help to interpret new microarray data. PMID- 20003505 TI - Developing 21st century accreditation standards for teaching hospitals: the Taiwan experience. AB - BACKGROUND: The purpose of this study is to establish teaching hospital accreditation standards anew with the hope that Taiwan's teaching hospitals can live up to the expectations of our society and ensure quality teaching. METHODS: The development process lasted two years, 2005-2006, and was separated into three stages. The first stage centered on leadership meetings and consensus building, the second on drafting the new standards with expert focus groups, and the third on a pilot study and subsequent revision. RESULTS: Our new teaching hospital accreditation standards have six categories and 95 standards as follows: educational resources (20 items), teaching and training plans and outcomes (42 items), research and results (9 items), development of clinical faculty and continuing education (8 items), academic exchanges and community education (8 items), and administration (8 items). CONCLUSIONS: The new standards have proven feasible and posed reasonable challenges in the pilot study. We hope the new standards will strengthen teaching and research, and improve the quality of hospital services at the same time. PMID- 20003506 TI - Unique case of esophageal rupture after a fall from height. AB - BACKGROUND: Traumatic ruptures of the esophagus are relatively rare. This condition is associated with high morbidity and mortality. Most traumatic ruptures occur after motor vehicle accidents. CASE PRESENTATION: We describe a unique case of a 23 year old woman that presented at our trauma resuscitation room after a fall from 8 meters. During physical examination there were no clinical signs of life-threatening injuries. She did however have a massive amount of subcutaneous emphysema of the chest and neck and pneumomediastinum. Flexible laryngoscopy revealed a lesion in the upper esophagus just below the level of the upper esophageal sphincter. Despite preventive administration of intravenous antibiotics and nutrition via a nasogastric tube, the patient developed a cervical abscess, which drained spontaneously. Normal diet was gradually resumed after 2.5 weeks and the patient was discharged in a reasonable condition 3 weeks after the accident. CONCLUSIONS: This case report presents a high cervical esophageal rupture without associated local injuries after a fall from height. PMID- 20003507 TI - Atomoxetine for the treatment of attention-deficit/hyperactivity disorder (ADHD) in children with ADHD and dyslexia. AB - BACKGROUND: The objective of this study was to assess the effects of atomoxetine on treating attention-deficit/hyperactivity disorder (ADHD), on reading performance, and on neurocognitive function in youth with ADHD and dyslexia (ADHD+D). METHODS: Patients with ADHD (n = 20) or ADHD+D (n = 36), aged 10-16 years, received open-label atomoxetine for 16 weeks. Data from the ADHD Rating Scale-IV (ADHDRS-IV), Kaufman Test of Educational Achievement (K-TEA), Working Memory Test Battery for Children (WMTB-C), and Life Participation Scale for ADHD Child Version (LPS-C) were assessed. RESULTS: Atomoxetine demonstrated significant improvement for both groups on the ADHDRS-IV, LPS-C, and K-TEA reading comprehension standard and composite scores. K-TEA spelling subtest improvement was significant for the ADHD group, whereas the ADHD+D group showed significant reading decoding improvements. Substantial K-TEA reading and spelling subtest age equivalence gains (in months) were achieved for both groups. The WMTB C central executive score change was significantly greater for the ADHD group. Conversely, the ADHD+D group showed significant phonological loop score enhancement by visit over the ADHD group. Atomoxetine was well tolerated, and commonly reported adverse events were similar to those previously reported. CONCLUSIONS: Atomoxetine reduced ADHD symptoms and improved reading scores in both groups. Conversely, different patterns and magnitude of improvement in working memory component scores existed between ADHD and ADHD+D patients. Though limited by small sample size, group differences in relation to the comparable changes in improvement in ADHD symptoms could suggest that brain systems related to the therapeutic benefit of atomoxetine in reducing ADHD symptoms may be different in individuals with ADHD+D and ADHD without dyslexia. TRIAL REGISTRATION: CLINICAL TRIAL REGISTRY: ClinicalTrials.gov: NCT00191048. PMID- 20003509 TI - The reproducibility of measurements of intramuscular magnesium concentrations and muscle oxidative capacity using 31P MRS. AB - 31P magnetic resonance spectroscopy (31P MRS) has been used to measure intramuscular magnesium concentrations and muscle metabolism. Abnormal intramuscular magnesium has been reported in several patient populations with suspected metabolic disorders. The purpose of this study was to evaluate our ability to measure intramuscular magnesium and muscle metabolism in the quadriceps muscles of healthy subjects, and to test whether these measurements were influenced by prior exercise. Twelve normal, healthy male volunteers were tested in a 3 Tesla magnet on four separate days. Resting [Mg2+] was calculated from the heights and frequency shifts of the phosphate, phosphocreatine and ATP peaks. Phosphocreatine (PCr) recovery kinetics were measured after 30-39 second bouts of isometric exercise. Thirty minutes prior to the 3rd test session the subjects completed a 2 hour treadmill walk at 40-60% of heart rate reserve. Resting [Mg2+] averaged 0.388 mM and had an interclass correlation coefficient between days (ICC) of 0.352. The mean end exercise PCr was 47.6% and the mean end exercise pH was 6.97. PCr recovery averaged 39 seconds (p = 0.892) and had an ICC of 0.819. Prior long duration exercise did not produce significant alterations in either PCr recovery kinetics or intracellular magnesium levels (p = 0.440). In conclusion, the reproducibility of Resting [Mg2+] was less than that of PCr recovery measurements, and may reflect the sensitivity of these measurements to phasing errors. In addition, prior exercise is unlikely to alter measurements of resting metabolites or muscle metabolism suggesting that rigorous control of physical activity prior to metabolic testing is unnecessary. PMID- 20003508 TI - Adolescents' wellbeing and functioning: relationships with parents' subjective general physical and mental health. AB - BACKGROUND: This study aimed at examining the relationship between parental subjective health status and adolescents' health-related quality of life (HRQoL) as well as the role of gender, socioeconomic status, presence of chronic health care needs and social support on the above interaction. METHODS: Questionnaires were administered to a Greek nation-wide random sample of adolescents (N = 1,194) aged 11-18 years and their parents (N = 973) in 2003. Adolescents' and parents' status was assessed, together with reports of socio-economic status and level of social support. Various statistical tests were used to determine the extent to which these variables were related to each other. RESULTS AND DISCUSSION: Parental subjective mental health status was significantly correlated with adolescents' better physical and psychological wellbeing, moods and emotions, parent-child relationships, school environment and financial resources. Parental subjective physical health status was strongly associated with more positive adolescents' self-perception. Adolescents' male gender, younger age, absence of chronic health care needs, high social support, and higher family income were positively associated with better HRQoL. CONCLUSIONS: This study reinforces the importance of parental subjective health status, along with other variables, as a significant factor for the adolescents' HRQoL. PMID- 20003510 TI - Comparative analysis of slot dimension in lingual bracket systems. AB - BACKGROUND: Orthodontic treatment with fixed appliances requires - among others - the correct clinical expression of torque, which depends on the precise fitting of archwire and slot. Especially in the lingual technique torque problems become clinically more evident than in labial appliances also with respect to the vertical alignment of teeth due to different distances from the center of resistance. The purpose of the present study was to compare the preciseness of slot dimensions of different lingual bracket systems. METHODS: Three lingual bracket systems were included in the study (7th Generation and STb, Ormco, Glendora, CA, USA; Incognito, TOP-Service/3 M Unitek, Monrovia, CA, USA). Non destructive analysis of vertical slot dimensions was performed using precision pin gauges (Azurea, Belprahon, Switzerland) that were tapered in increments of 0.002 mm (0.00008 inch). The sizes of 240 incisor and canine brackets were measured per system (total: 720). Data were compared using one-way ANOVA. A p value < 0.05 was considered statistically significant. RESULTS: Average slot dimensions were 0.467 mm +/- 0.007 mm (0.0184 inch +/- 0.0003 inch) for the 7th Generation bracket system, 0.466 mm +/- 0.004 mm (0.0183 inch +/- 0.0001) inch for the STb bracket system and 0.459 mm +/- 0.004 mm (0.0181 inch +/- 0.0001) inch for the Incognito bracket system. Differences between systems were statistically significant (p < 0.05). CONCLUSIONS: The analyzed bracket systems for lingual treatment exhibited significant differences in slot dimension that will clinically result in torque play. These aspects must be considered in lingual orthodontic treatment. PMID- 20003511 TI - Autologous bone graft versus demineralized bone matrix in internal fixation of ununited long bones. AB - BACKGROUND: Non-unions are severe complications in orthopaedic trauma care and occur in 10% of all fractures. The golden standard for the treatment of ununited fractures includes open reduction and internal fixation (ORIF) as well as augmentation with autologous-bone-grafting. However, there is morbidity associated with the bone-graft donor site and some patients offer limited quantity or quality of autologous-bone graft material. Since allogene bone-grafts are introduced on the market, this comparative study aims to evaluate healing characteristics of ununited bones treated with ORIF combined with either iliac crest-autologous-bone-grafting (ICABG) or demineralized-bone-matrix (DBM). METHODS AND RESULTS: From 2000 to 2006 out of sixty-two consecutive patients with non-unions presenting at our Level I Trauma Center, twenty patients had ununited diaphyseal fractures of long bones and were treated by ORIF combined either by ICABG- (n = 10) or DBM-augmentation (n = 10). At the time of index-operation, patients of the DBM-group had a higher level of comorbidity (ASA-value: p = 0.014). Mean duration of follow-up was 56.6 months (ICABG-group) and 41.2 months (DBM-group). All patients were clinically and radiographically assessed and adverse effects related to bone grafting were documented. The results showed that two non-unions augmented with ICABG failed osseous healing (20%) whereas all non unions grafted by DBM showed successful consolidation during the first year after the index operation (p = 0.146). No early complications were documented in both groups but two patients of the ICABG-group suffered long-term problems at the donor site (20%) (p = 0.146). Pain intensity were comparable in both groups (p = 0.326). However, patients treated with DBM were more satisfied with the surgical procedure (p = 0.031). CONCLUSION: With the use of DBM, the costs for augmentation of the non-union-site are more expensive compared to ICABG (calculated difference: 160 euro/case). Nevertheless, this study demonstrated that the application of DBM compared to ICABG led to an advanced outcome in the treatment of non-unions and simultaneously to a decreased quantity of adverse effects. Therefore we conclude that DBM should be offered as an alternative to ICABG, in particular to patients with elevated comorbidity and those with limited availability or reduced quality of autologous-bone graft material. PMID- 20003512 TI - Antenatal and postnatal corticosteroid and resuscitation induced lung injury in preterm sheep. AB - BACKGROUND: Initiation of ventilation using high tidal volumes in preterm lambs causes lung injury and inflammation. Antenatal corticosteroids mature the lungs of preterm infants and postnatal corticosteroids are used to treat bronchopulmonary dysplasia. OBJECTIVE: To test if antenatal or postnatal corticosteroids would decrease resuscitation induced lung injury. METHODS: 129 d gestational age lambs (n = 5-8/gp; term = 150 d) were operatively delivered and ventilated after exposure to either 1) no medication, 2) antenatal maternal IM Betamethasone 0.5 mg/kg 24 h prior to delivery, 3) 0.5 mg/kg Dexamethasone IV at delivery or 4) Cortisol 2 mg/kg IV at delivery. Lambs then were ventilated with no PEEP and escalating tidal volumes (VT) to 15 mL/kg for 15 min and then given surfactant. The lambs were ventilated with VT 8 mL/kg and PEEP 5 cmH20 for 2 h 45 min. RESULTS: High VT ventilation caused a deterioration of lung physiology, lung inflammation and injury. Antenatal betamethasone improved ventilation, decreased inflammatory cytokine mRNA expression and alveolar protein leak, but did not prevent neutrophil influx. Postnatal dexamethasone decreased pro-inflammatory cytokine expression, but had no beneficial effect on ventilation, and postnatal cortisol had no effect. Ventilation increased liver serum amyloid mRNA expression, which was unaffected by corticosteroids. CONCLUSIONS: Antenatal betamethasone decreased lung injury without decreasing lung inflammatory cells or systemic acute phase responses. Postnatal dexamethasone or cortisol, at the doses tested, did not have important effects on lung function or injury, suggesting that corticosteroids given at birth will not decrease resuscitation mediated injury. PMID- 20003513 TI - Transcribed-Ultra Conserved Region expression is associated with outcome in high risk neuroblastoma. AB - BACKGROUND: Neuroblastoma is the most common, pediatric, extra-cranial, malignant solid tumor. Despite multimodal therapeutic protocols, outcome for children with a high-risk clinical phenotype remains poor, with long-term survival still less than 40%. Hereby, we evaluated the potential of non-coding RNA expression to predict outcome in high-risk, stage 4 neuroblastoma. METHODS: We analyzed expression of 481 Ultra Conserved Regions (UCRs) by reverse transcription quantitative real-time PCR and of 723 microRNAs by microarrays in 34 high-risk, stage 4 neuroblastoma patients. RESULTS: First, the comparison of 8 short- versus 12 long-term survivors showed that 54 UCRs were significantly (P < 0.0491) over expressed in the former group. For 48 Ultra Conserved Region (UCRs) the expression levels above the cut-off values defined by ROC curves were strongly associated with good-outcome (OS: 0.0001

0 to <10, >or=10 years) in industry or occupation. RESULTS: Significantly increased risk of UBC was observed among waiters and bartenders (OR 2.87; 95% CI 1.05 to 7.72) and occupations related to medicine and health (OR 2.17; 95% CI 1.21 to 3.92), agricultural production, livestock and animal specialties (OR 1.90; 95% CI 1.03 to 3.49), electrical assembly, installation and repair (OR 1.69; 95% CI 1.07 to 2.65), communications (OR 1.74; 95% CI 1.00 to 3.01), and health services (OR 1.58; 95% CI 1.02 to 2.44). For these occupations we also observed a significant excess risk of UBC for long-term work (i.e. >or=10 years), with the exception of waiters and bartenders. Employment for 10 years or more was associated with increased risk of UBC in general farmers (OR 9.58; 95% CI 2.18 to 42.05), agricultural production of crops (OR 3.36; 95% CI 1.10 to 10.27), occupations related to bench working (OR 4.76; 95% CI 1.74 to 13.01), agricultural, fishery, forestry & related (OR 4.58; 95% CI 1.97 to 10.65), transportation equipment (OR 2.68; 95% CI 1.03 to 6.97), and structural work (OR 1.85; 95% CI 1.16 to 2.95). CONCLUSIONS: This study provides evidence of increased risk of UBC for occupations that were previously reported as at-risk. Workers in several occupation and industry groups have a significantly higher risk of UBC, particularly when duration of employment is 10 years or more. PMID- 20003538 TI - Unrestricted somatic stem cells from human umbilical cord blood grow in serum free medium as spheres. AB - BACKGROUND: Human umbilical cord blood-derived unrestricted somatic stem cells (USSCs), which are capable of multilineage differentiation, are currently under investigation for a number of therapeutic applications. A major obstacle to their clinical use is the fact that in vitro expansion is still dependent upon fetal calf serum, which could be a source of pathogens. In this study, we investigate the capacity of three different stem cell culture media to support USSCs in serum free conditions; HEScGRO, PSM and USSC growth medium ACF. Our findings demonstrate that USSCs do not grow in HEScGRO or PSM, but we were able to isolate, proliferate and maintain multipotency of three USSC lines in USSC growth medium ACF. RESULTS: For the first one to three passages, cells grown in USSC growth medium ACF proliferate and maintain their morphology, but with continued passaging the cells form spherical cell aggregates. Upon dissociation of spheres, cells continue to grow in suspension and form new spheres. Dissociated cells can also revert to monolayer growth when cultured on extracellular matrix support (fibronectin or gelatin), or in medium containing fetal calf serum. Analysis of markers associated with pluripotency (Oct4 and Sox2) and differentiation (FoxA2, Brachyury, Goosecoid, Nestin, Pax6, Gata6 and Cytokeratin 8) confirms that cells in the spheres maintain their gene expression profile. The cells in the spheres also retain the ability to differentiate in vitro to form cells representative of the three germline layers after five passages. CONCLUSIONS: These data suggest that USSC growth medium ACF maintains USSCs in an undifferentiated state and supports growth in suspension. This is the first demonstration that USSCs can grow in a serum- and animal component-free medium and that USSCs can form spheres. PMID- 20003539 TI - Malaria control in Timor-Leste during a period of political instability: what lessons can be learned? AB - BACKGROUND: Malaria is a major global health problem, often exacerbated by political instability, conflict, and forced migration. OBJECTIVES: To examine the impact of political upheaval and population displacement in Timor-Leste (2006) on malaria in the country. METHOD: Case study approach drawing on both qualitative and quantitative methods including document reviews, in-depth interviews, focus group discussions, site visits and analysis of routinely collected data. FINDINGS: The conflict had its most profound impact on Dili, the capital city, in which tens of thousands of people were displaced from their homes. The conflict interrupted routine malaria service programs and training, but did not lead to an increase in malaria incidence. Interventions covering treatment, insecticide treated nets (ITN) distribution, vector control, surveillance and health promotion were promptly organized for internally displaced people (IDPs) and routine health services were maintained. Vector control interventions were focused on IDP camps in the city rather than on the whole community. The crisis contributed to policy change with the introduction of Rapid Diagnostic Tests and artemether-lumefantrine for treatment. CONCLUSIONS: Although the political crisis affected malaria programs there were no outbreaks of malaria. Emergency responses were quickly organized and beneficial long term changes in treatment and diagnosis were facilitated. PMID- 20003540 TI - Associations of physical and mental health problems with chronic cough in a representative population cohort. AB - BACKGROUND: Although chronic cough is a common problem in clinical practice, data on the prevalence and characteristics of cough in the general population are scarce. Our aim was to determine the prevalence of chronic cough that is not associated with diagnosed respiratory conditions and examine the impact on health status and psychological health, in a representative adult population cohort METHODS: North West Adelaide Health Study (n stage 1 = 4060, stage 2 = 3160) is a representative population adult cohort. Clinical assessment included spirometry, anthropometry and skin tests. Questionnaires assessed demographics, lifestyle risk factors, quality of life, mental health and respiratory symptoms, doctor diagnosed conditions and medication use. RESULTS: Of the 3355 people without identified lung disease at baseline, 18.2% reported chronic cough. In multiple logistic regression models, at follow-up, dry chronic cough without sputum production was significantly more common in males (OR 1.5, 95% CI 1.1, 1.9), current smokers (OR 4.9, 95% CI 3.4, 7.2), obesity (OR 1.9, 95% CI 1.3, 2.9), use of ACE inhibitors (OR 1.8, 95% CI 1.1, 2.9), severe mental health disturbance (OR 2.1, 95% CI 1.4, 3.1) and older age (40-59 years OR 1.7 95% CI 1.2, 2.4; > or = 60 years OR 2.1 95% CI 1.3, 3.5). Among non-smokers only, all cough was significantly more common in men, those with severe mental health disturbance and obesity. CONCLUSIONS: Chronic cough is a major cause of morbidity. Attention to cough is indicated in patients with obesity, psychological symptoms or smokers. Inquiring about cough in those with mental health problems may identify reversible morbidity. PMID- 20003541 TI - The hemolytic and cytolytic activities of Serratia marcescens phospholipase A (PhlA) depend on lysophospholipid production by PhlA. AB - BACKGROUND: Serratia marcescens is a gram-negative bacterium and often causes nosocomial infections. There have been few studies of the virulence factors of this bacterium. The only S. marcescens hemolytic and cytotoxic factor reported, thus far, is the hemolysin ShlA. RESULTS: An S. marcescens shlAB deletion mutant was constructed and shown to have no contact hemolytic activity. However, the deletion mutant retained hemolytic activity on human blood agar plates, indicating the presence of another S. marcescens hemolytic factor. Functional cloning of S. marcescens identified a phospholipase A (PhlA) with hemolytic activity on human blood agar plates. A phlAB deletion mutant lost hemolytic activity on human blood agar plates. Purified recombinant PhlA hydrolyzed several types of phospholipids and exhibited phospholipase A1 (PLA1), but not phospholipase A2 (PLA2), activity. The cytotoxic and hemolytic activities of PhlA both required phospholipids as substrates. CONCLUSION: We have shown that the S. marcescens phlA gene produces hemolysis on human blood agar plates. PhlA induces destabilization of target cell membranes in the presence of phospholipids. Our results indicated that the lysophospholipids produced by PhlA affected cell membranes resulting in hemolysis and cell death. PMID- 20003542 TI - A simple method for estimating genetic diversity in large populations from finite sample sizes. AB - BACKGROUND: Sample size is one of the critical factors affecting the accuracy of the estimation of population genetic diversity parameters. Small sample sizes often lead to significant errors in determining the allelic richness, which is one of the most important and commonly used estimators of genetic diversity in populations. Correct estimation of allelic richness in natural populations is challenging since they often do not conform to model assumptions. Here, we introduce a simple and robust approach to estimate the genetic diversity in large natural populations based on the empirical data for finite sample sizes. RESULTS: We developed a non-linear regression model to infer genetic diversity estimates in large natural populations from finite sample sizes. The allelic richness values predicted by our model were in good agreement with those observed in the simulated data sets and the true allelic richness observed in the source populations. The model has been validated using simulated population genetic data sets with different evolutionary scenarios implied in the simulated populations, as well as large microsatellite and allozyme experimental data sets for four conifer species with contrasting patterns of inherent genetic diversity and mating systems. Our model was a better predictor for allelic richness in natural populations than the widely-used Ewens sampling formula, coalescent approach, and rarefaction algorithm. CONCLUSIONS: Our regression model was capable of accurately estimating allelic richness in natural populations regardless of the species and marker system. This regression modeling approach is free from assumptions and can be widely used for population genetic and conservation applications. PMID- 20003543 TI - Immunohistochemical and other prognostic factors in B cell non Hodgkin lymphoma patients, Kampala, Uganda. AB - BACKGROUND: Non Hodgkin lymphomas are the most common lymphomas in Uganda. Recent studies from developed countries have shown differences in survival for the different immunophenotypes. Such studies are lacking in Africa where diagnosis is largely dependent on morphology alone. We report immunohistochemical and other prognostic factors in B cell non Hodgkin lymphoma patients in Kampala, Uganda. METHODS: Non Hodgkin lymphoma tissue blocks from the archives of the Department of Pathology, Makerere University College of Health Sciences, Kampala, Uganda, from 1991-2000, were sub typed using haematoxylin and eosin, Giemsa as well as immunohistochemistry. Using tissue micro array, 119 biopsies were subjected to: CD3, CD5, CD10, CD20, CD23, CD30, CD38, CD79a, CD138, Bcl-6, Bcl-2, IRTA-1, MUM1/IRF4, Bcl-1/cyclin D1, TdT, ALKc, and Ki-67/Mib1. Case notes were retrieved for: disease stage, chemotherapy courses received and retrospective follow up was done for survival. RESULTS: Non Hodgkin B cell lymphomas comprised of Burkitt lymphoma [BL] (95/119) diffuse large B cell lymphoma (19/119), mantle cell lymphoma (4/119) and precursor B lymphoblastic lymphoma (1/119). For Burkitt lymphoma, good prognosis was associated with receiving chemotherapy, female gender and CD30 positivity. Only receiving chemotherapy remained significant after Cox regression analysis. Diffuse large B cell lymphomas with activated germinal centre B cell (GCB) pattern (CD10+/-, BCL-6+/-, MUM+/-, CD138+/-) had better survival (98.4 months; 95% CI 89.5 -107.3) than the others (57.3 months; 95% CI 35.5 - 79.0) p = 0.027 (log rank test). CONCLUSIONS: Activated GCB diffuse large B cell lymphoma had a better prognosis than the others. For Burkitt lymphoma, not receiving chemotherapy carried a poor prognosis. Availability of chemotherapy in this resource limited setting is critical for survival of lymphoma patients. PMID- 20003544 TI - CHESS (CgHExpreSS): a comprehensive analysis tool for the analysis of genomic alterations and their effects on the expression profile of the genome. AB - BACKGROUND: Genomic alterations frequently occur in many cancer patients and play important mechanistic roles in the pathogenesis of cancer. Furthermore, they can modify the expression level of genes due to altered copy number in the corresponding region of the chromosome. An accumulating body of evidence supports the possibility that strong genome-wide correlation exists between DNA content and gene expression. Therefore, more comprehensive analysis is needed to quantify the relationship between genomic alteration and gene expression. A well-designed bioinformatics tool is essential to perform this kind of integrative analysis. A few programs have already been introduced for integrative analysis. However, there are many limitations in their performance of comprehensive integrated analysis using published software because of limitations in implemented algorithms and visualization modules. RESULTS: To address this issue, we have implemented the Java-based program CHESS to allow integrative analysis of two experimental data sets: genomic alteration and genome-wide expression profile. CHESS is composed of a genomic alteration analysis module and an integrative analysis module. The genomic alteration analysis module detects genomic alteration by applying a threshold based method or SW-ARRAY algorithm and investigates whether the detected alteration is phenotype specific or not. On the other hand, the integrative analysis module measures the genomic alteration's influence on gene expression. It is divided into two separate parts. The first part calculates overall correlation between comparative genomic hybridization ratio and gene expression level by applying following three statistical methods: simple linear regression, Spearman rank correlation and Pearson's correlation. In the second part, CHESS detects the genes that are differentially expressed according to the genomic alteration pattern with three alternative statistical approaches: Student's t-test, Fisher's exact test and Chi square test. By successive operations of two modules, users can clarify how gene expression levels are affected by the phenotype specific genomic alterations. As CHESS was developed in both Java application and web environments, it can be run on a web browser or a local machine. It also supports all experimental platforms if a properly formatted text file is provided to include the chromosomal position of probes and their gene identifiers. CONCLUSIONS: CHESS is a user-friendly tool for investigating disease specific genomic alterations and quantitative relationships between those genomic alterations and genome-wide gene expression profiling. PMID- 20003545 TI - Perceived body image in men and women with type 2 diabetes mellitus: correlation of body mass index with the figure rating scale. AB - BACKGROUND: Body mass index (BMI) is often used as an objective surrogate estimate of body fat. Increased BMI is directly associated with an increase in metabolic disease, such as type 2 diabetes mellitus (T2DM). The Stunkard Figure Rating Scale (FRS) is a subjective measure of body fat, and self-perceptions of body image conceivably impact the development and treatment of T2DM. This study examined the self-perception of body image to various levels of BMI among those with T2DM. METHODS: Respondents (n = 13,887) to the US Study to Help Improve Early evaluation and management of risk factors Leading to Diabetes (SHIELD) 2006 survey self-reported their weight and height for BMI calculation. On the gender specific Stunkard FRS, respondents selected the figure most closely resembling their body image. Spearman correlation was computed between perceived body image and BMI for men and women separately. Student's t-test analysis compared the mean BMI differences between respondents with and without T2DM. RESULTS: Men with T2DM did not significantly differ from men without diabetes mellitus in mean BMI per body image figure except at the extremes in body figures. Women with T2DM had a significantly higher BMI for the same body figure compared with women without diabetes mellitus for most figures (p < 0.05). CONCLUSIONS: Individuals, particularly women, with T2DM may differ in their perception of body image compared with those without diabetes mellitus. It is unclear if these perceived differences increase the risk of T2DM, or if the diagnosis of T2DM alters body image perceptions. PMID- 20003546 TI - Optimising intraperitoneal gentamicin dosing in peritoneal dialysis patients with peritonitis (GIPD) study. AB - BACKGROUND: Antibiotics are preferentially delivered via the peritoneal route to treat peritonitis, a major complication of peritoneal dialysis (PD), so that maximal concentrations are delivered at the site of infection. However, drugs administered intraperitoneally can be absorbed into the systemic circulation. Drugs excreted by the kidneys accumulate in PD patients, increasing the risk of toxicity. The aim of this study is to examine a model of gentamicin pharmacokinetics and to develop an intraperitoneal drug dosing regime that maximises bacterial killing and minimises toxicity. METHODS/DESIGN: This is an observational pharmacokinetic study of consecutive PD patients presenting to the Royal Brisbane and Women's Hospital with PD peritonitis and who meet the inclusion criteria. Participants will be allocated to either group 1, if anuric as defined by urine output less than 100 ml/day, or group 2: if non-anuric, as defined by urine output more than 100 ml/day. Recruitment will be limited to 15 participants in each group. Gentamicin dosing will be based on the present Royal Brisbane & Women's Hospital guidelines, which reflect the current International Society for Peritoneal Dialysis Peritonitis Treatment Recommendations. The primary endpoint is to describe the pharmacokinetics of gentamicin administered intraperitoneally in PD patients with peritonitis based on serial blood and dialysate drug levels. DISCUSSION: The study will develop improved dosing recommendations for intraperitoneally administered gentamicin in PD patients with peritonitis. This will guide clinicians and pharmacists in selecting the most appropriate dosing regime of intraperitoneal gentamicin to treat peritonitis. TRIAL REGISTRATION: ACTRN12609000446268. PMID- 20003547 TI - Association of a de novo 16q copy number variant with a phenotype that overlaps with Lenz microphthalmia and Townes-Brocks syndromes. AB - BACKGROUND: Anophthalmia and microphthalmia are etiologically and clinically heterogeneous. Lenz microphthalmia is a syndromic form that is typically inherited in an X-linked pattern, though the causative gene mutation is unknown. Townes-Brocks syndrome manifests thumb anomalies, imperforate anus, and ear anomalies. We present a 13-year-old boy with a syndromic microphthalmia phenotype and a clinical diagnosis of Lenz microphthalmia syndrome. CASE PRESENTATION: The patient was subjected to clinical and molecular evaluation, including array CGH analysis. The clinical features included left clinical anophthalmia, right microphthalmia, anteriorly placed anus with fistula, chordee, ventriculoseptal defect, patent ductus arteriosus, posteriorly rotated ears, hypotonia, growth retardation with delayed bone age, and mental retardation. The patient was found to have an approximately 5.6 Mb deletion of 16q11.2q12.1 by microarray based comparative genomic hybridization, which includes the SALL1 gene, which causes Townes-Brocks syndrome. CONCLUSIONS: Deletions of 16q11.2q12.2 have been reported in several individuals, although those prior reports did not note microphthalmia or anophthalmia. This region includes SALL1, which causes Townes-Brocks syndrome. In retrospect, this child has a number of features that can be explained by the SALL1 deletion, although it is not clear if the microphthalmia is a rare feature of Townes-Brocks syndrome or caused by other mechanisms. These data suggest that rare copy number changes may be a cause of syndromic microphthalmia allowing a personalized genomic medicine approach to the care of patients with these aberrations. PMID- 20003548 TI - Genetic diversity of Ophiocordyceps sinensis, a medicinal fungus endemic to the Tibetan Plateau: implications for its evolution and conservation. AB - BACKGROUND: Ophiocordyceps sinensis (syn. Cordyceps sinensis), endemic to alpine regions on the Tibetan plateau, is one of the most valuable medicinal fungi in the world. Huge commercial demand has led to excessive harvest and a dramatic decline in its numbers. The diversity of terrains and climates on the Tibetan Plateau and the broad insect host range (more than 50 species in the family Hepialidae) may have resulted in substantial intraspecific genetic diversity for this fungus. The objective of this study was to evaluate the population distribution of O. sinensis from geographically diverse regions of the Tibetan Plateau based on nrDNA ITS and MAT1-2-1 gene sequences. Understanding of the genetic diversity and genesis of O. sinensis will provide important information for the evolution and conservation of this fungus. RESULTS: Significant sequence variations in the ITS and MAT1-2-1 genes (27 and 23 informative sites, eight and seven haplotypes, respectively) were observed. Phylogenetic analysis based on ITS sequences, MAT1-2-1 sequences, or their combined data set, clustered isolates from northern regions in one clade (clade I), whereas isolates from southern regions were dispersed in all four clades (clade I-IV). Single-strand conformation polymorphism (SSCP) analyses of 2639 ITS clones from seven samples revealed 91 different SSCP patterns that were subsequently sequenced. ITS heterogeneity was found in XZ-LZ07-H1 (Nyingchi population), and 17 informative sites and five haplotypes were detected from 15 clones. The five haplotypes clustered into three clades (clade I, II, and IV). CONCLUSIONS: Significant genetic divergence in O. sinensis was observed and the genetic diversification was greater among southern isolates than that among northern isolates. The polymorphism of nrDNA ITS sequences suggested that O. sinensis spread from a center of origin (the Nyingchi District) to southern regions and subsequently to northern areas. These results suggest that southern populations are important reservoirs of genetic diversity and should be taken into account in conservation programs. PMID- 20003550 TI - Survival of Ascaris eggs and hygienic quality of human excreta in Vietnamese composting latrines. AB - BACKGROUND: For centuries farmers in Vietnam have fertilized their fields with human excreta collected directly from their household latrines. Contrary to the official guideline of six-month storage, the households usually only store human excreta for three to four months before use, since this is the length of time that farmers have available to produce fertilizer between two cropping seasons. This study aimed to investigate whether hygienically safe fertilizer could be produced in the latrines within this period of time. METHODS: By inoculating eggs of the helminth parasite indicator Ascaris suum into heaps of human excreta, a die-off experiment was conducted under conditions similar to those commonly used in Vietnamese latrines. Half a ton of human excreta was divided into five heaps containing increasing concentrations of lime from 0% to 11%. RESULTS: Regardless of the starting pH, which varied from 9.4 to 11.6, a >99% die-off of eggs was obtained after 105 to 117 days of storage for all lime concentrations and 97% of eggs were non-viable after 88 days of storage. The most critical parameter found to determine the die-off process was the amount of ammonia (urine) in the excreta which indicates that longer storage periods are needed for parasite egg die-off if urine is separated from the excreta. CONCLUSION: By inactivating >99% of all A. suum eggs in human excreta during a storage period of only three months the commonly used Double Vault Composting (DVC) latrine, in which urine is not separated, could therefore potentially provide a hygienic acceptable fertilizer. PMID- 20003551 TI - [Diagnostic image. A woman with a vulvar swelling]. AB - A 25-year-old woman presented with a tumour above the labia minora, due to an epidermal cyst. PMID- 20003552 TI - [Colorectal cancer screening: advice from the Health Council of the Netherlands]. AB - There is sufficient evidence that a colorectal cancer (CRC) screening programme is feasible, acceptable and cost-effective amongst the Dutch population. This is the conclusion of the Health Council of the Netherlands in an advisory report to the Minister of Health, Welfare and Sport. It also concluded that an immunochemical faecal occult blood test (iFOBT) would be the obvious screening method. The recommended target group is men and women aged 55-75. An evaluation of the randomized pilot trials in the regions of Nijmegen, Amsterdam, and Rotterdam predicted a participation rate of 60%. Based on the analyses from a microsimulation model, biennial screening for CRC with iFOBT and an uptake rate of 60% could prevent 1400 deaths from CRC per annum, at a cost of 2200 euros per years of life saved. PMID- 20003553 TI - [Negative pressure therapy for surgical wounds]. AB - In surgery, negative pressure wound therapy (NPWT) is applied for thoracic, abdominal, and extremity wounds, particularly if there is traumatic tissue loss, if primary wound closure is not possible or if the wound has to be left open or reopened because of an infection. The NPWT applications are, however, running ahead of scientific evidence to support their use. Although there is plenty of literature available, very little convincing evidence exists based on well conducted, randomised trials. NPWT can be applied to thoracic wounds (usually due to deep sternal wound infections) to reduce treatment duration. NPWT also appears useful if primary closure of the abdomen is not possible. For traumatic extremity wounds, NPWT can be helpful as a practical temporary wound cover. Possible disadvantages of NPWT are skin irritation, painful dressing changes, entero atmospheric fistulae, and the risk of bleeding in patients using anticoagulants. The effect of NPWT on wound healing, infection, mortality, and costs remains unclear. To obtain convincing evidence, we call for greater NPWT application in a research context. PMID- 20003554 TI - [Acute pelvic fractures]. AB - Two patients were involved in a high-energy trauma that resulted in an unstable pelvic fracture. The first patient, a 51-year-old woman, was trapped between two boats. At our hospital she received a traumatic pelvic orthotic device (T-POD), followed by angiography with embolisation. Three days post-trauma she went to the operating room for definitive surgical treatment of her Tile C pelvic fracture. The second patient, a 19-year-old man, was in a car that collided with a tree. He also received a T-POD, but remained haemodynamically unstable. He went to the operating room for damage control surgery, followed by an angiography with embolisation. Two days later, definitive surgical treatment of the Tile C pelvic fracture took place. These cases illustrate the variety of possible treatments for patients with unstable pelvic fractures. Choice of treatment depends on the character of the attending injuries and the haemodynamic situation. Fast consultation should take place with a hospital experienced in the initial care and treatment of unstable pelvic fractures. PMID- 20003555 TI - [Colorectal cancer screening: CT colonography]. AB - In the Netherlands, the introduction of screening for colorectal cancer is being considered. CT colonography is an accurate technique for the detection of colorectal cancer and its precursors in symptomatic patients and for screening. Radiation exposure in CT colonography is limited and is no major drawback for the use of CT colonography for screening. Aspects important to screening with CT colonography have not been studied yet, such as participation rate and cost effectiveness. MR colonography for screening has been studied little. PMID- 20003556 TI - [Diagnosis of colorectal tumours in daily clinical practice: comparison of colonoscopy and sigmoidoscopy]. AB - OBJECTIVE: To map the locations of advanced colorectal neoplasia in patients referred for colonoscopy or sigmoidoscopy and to compare the yield of advanced neoplasia and the distribution of advanced neoplasia per indication for endoscopy. DESIGN: Observational. METHOD: In a multicentre survey in North Holland, the Netherlands (n = 18 hospitals), data on all colonoscopies and sigmoidoscopies performed during a three-month period in 2005 were analyzed. The locations and the histological features of all colonic neoplasia and the indications for endoscopy were recorded. Advanced neoplasm was defined as adenoma >or=10 mm in size, an adenoma with any villous features, or high-grade dysplasia or adenocarcinoma. RESULTS: A total of 4623 patients underwent a total colonoscopy and 3004 patients underwent sigmoidoscopy. The prevalence of advanced neoplasia was 13% on colonoscopy and 6% on sigmoidoscopy. Of the advanced neoplasia found on colonoscopy, 67% were located in the distal colon and 33% in the proximal colon. Of the patients with advanced neoplasia in the proximal colon (n = 228), 51% had no abnormalities in the distal colon. The percentage of advanced neoplasia in the proximal colon varied from 23% in patients younger than 50 years to 41% in patients aged 80 years and older. Depending on the indication for endoscopy, the prevalence of advanced neoplasia in the proximal colon varied from 11-57%. CONCLUSION: Of the advanced colorectal neoplasms 33% were located in the proximal colon. With increasing age, a shift in tumour localization occurs from distal to proximal in the colon. Colonoscopy is the preferred method for the endoscopic diagnosing of colorectal neoplasia. PMID- 20003557 TI - [Unexplained physical symptoms successfully treated by symptom-based psychotherapy]. AB - A 43-year-old woman suffered severe weight loss due to unexplained vomiting. No physical cause was detected. She was treated with hypnotherapy, cognitive therapy and a gradual increase in food with the help of a dietician. The vomiting stopped and the patient was able to eat again. For patients with unexplained physical symptoms, it is often difficult to understand that a psychological component is involved. This psychological component should not be emphasized. It is more important to tell the patient that there is no explanation for their problem, but that there is a possible treatment: psychotherapy. PMID- 20003558 TI - [Shared Medical Appointments: experiences of patients and care givers]. AB - OBJECTIVE: To examine the characteristics of patients participating in a shared medical appointment (SMA); to what extent SMAs fulfil patients' needs; to evaluate if patients are better informed after an SMA and how they evaluate meeting other patients at an SMA, and how health professionals experience SMAs. DESIGN: Descriptive. METHOD: Fourteen multidisciplinary teams participated in the study. The Dutch Institute for Healthcare Improvement (CBO) trained them in how to conduct an SMA. The experiences of patients participating in an SMA were compared with the experiences of patients who visited the doctor individually. Patients completed questionnaires before and after the SMA (n = 83) or the individual visit (n = 158). RESULTS: Patients who participated in an SMA did not differ significantly from the regular patient population in either demographic characteristics or in their experience of health and care uptake. Patients participated in an SMA primarily to share experiences with fellow patients, to learn from others and to obtain more information. Both patients and health professionals (n = 53) reported that patients received more information during an SMA. Patients valued the contact with their fellow patients. SMA offered care givers a different experience and opportunities to learn. CONCLUSION: For patients and health professionals, SMAs seem indeed to have added value. More research is needed to understand the working mechanism and the long-term effects of participation in SMAs. PMID- 20003560 TI - [Liver rupture caused by migrating gallstones]. AB - Gallstones can work their way through different tissue layers by means of chronic irritation and the formation of abscesses. Here we present the case of a 74-year old man who, after having attacks of pain in the upper abdominal region for years, was admitted with signs of peritoneal irritation and haemodynamic shock. An emergency laparotomy revealed old blood, pus and a large hole in the frontal part of the liver. A loose gallstone was found in the cavity. Each step this gallstone had taken from the gallbladder to the peritoneal cavity could, with hindsight, be traced back to the clinical course of the disease. Liver abscesses and rupture are rare complications of cholelithiasis. PMID- 20003562 TI - [Preconceptual care should include looking at the effect of working conditions on pregnancy]. AB - Working conditions are amongst the exogenous factors that can lead to pregnancy complications. Attention within preconceptual care to the working conditions experienced by women who plan to become pregnant can change this, by influencing exposure to harmful working conditions. Until now, relatively little attention has been given to working conditions within preconceptual care, and the subject is often not discussed in daily midwifery practice. The most relevant literature on this topic is summarized in this article. Occupational exposure to certain chemicals, stressful work and nightshifts during early pregnancy increase the risk of spontaneous abortion. There is also an increased risk of congenital malformations if the mother has been exposed to solvents or pesticides during early pregnancy, or the father has been exposed to these substances before or shortly after conception. We conclude that the occupational physician should be involved in the preconceptual consultation in order to contribute to the prevention of pregnancy complications. PMID- 20003563 TI - [Visual loss as a sign of hypertension]. AB - A 26-year-old male patient had been suffering from a decreased visual acuity in both eyes for 3 weeks. This appeared to be due to malignant hypertension. The hypertension went unnoticed until papillary and macular oedema were detected during fundoscopy. Hypertension can develop at all ages and may give rise to visual complaints or even to loss of vision. Insufficient clinical awareness of the atypical manifestations of severe hypertension and of the differential diagnosis of loss of vision may lead to irreversible damage of organs, in this patient the left eye. Fundoscopy is sometimes indicated in patients with severe hypertension to assess damage to the eye and to decide whether the patient must be hospitalised. PMID- 20003564 TI - [Recognition of temporal lobe epilepsy in adults]. AB - Temporal lobe epilepsy has a subtle and diverse symptomatology, and therefore temporal lobe seizures can initially be misdiagnosed. Here we discuss 3 patients with this type of epilepsy, but with completely different presentations. The first, a 67-year-old woman suffered from episodes of confusion, and later she developed tonic-clonic seizures. The second patient, a 58-year-old man, had auras followed by impaired consciousness and oral automatisms and also developed a type of seizure resembling panic attacks. The third patient, a 65-year-old man, presented with isolated auras. A few years later he developed tonic-clonic seizures, and the diagnosis 'temporal lobe epilepsy' was made. This article discusses differences in presentation between limbic and neocortical temporal lobe epilepsy, as well as the results of EEG and MRI investigations. PMID- 20003565 TI - [Diagnostic image. A woman with a swollen tongue]. AB - A 46-year-old woman had a swollen tongue due to amyloid light chain (AL) amyloidosis caused by multiple myeloma. PMID- 20003566 TI - [Multiparametric MRI for prostate cancer screening]. AB - Two recent studies on prostate cancer screening gave conflicting results about the effects of screening on prostate cancer mortality. The current screening method of PSA determination in combination with transrectal ultrasonic biopsy leads to a large number of false positive results and overtreatment. A screening test is needed that reduces the number of unnecessary prostate biopsies and which discriminates between more and less aggressive forms of prostate cancer. Multiparametric MRI has a high specificity for prostate cancer detection and provides information about prostate cancer aggressiveness. PSA in combination with multiparametric MRI performed at 1.5 Tesla appears to be a fairly accurate screening test. Due to its high costs and limited availability, multiparametric MRI is not suitable as a primary screening test. However, it could serve as a subsequent screening test if the PSA has increased above threshold values. Using multiparametric MRI as a follow-up test during screening would provide more accurate biopsies, prevent unnecessary prostate biopsies and improve the characterization of prostate cancer. PMID- 20003567 TI - Role of intestinal Bifidobacterium pseudolongum in dietary fructo-oligosaccharide inhibition of 2,4-dinitrofluorobenzene-induced contact hypersensitivity in mice. AB - Strategies to manipulate the gut microbiota have been explored for preventing allergy development. We previously showed that dietary supplementation with fructo-oligosaccharide (FOS) reduced 2, 4-dinitrofluorobenzene (DNFB)-induced contact hypersensitivity (CHS) in BALB/c mice. Because the CHS response was negatively correlated with the number of faecal bifidobacteria, particularly Bifidobacterium pseudolongum, the present study aimed to examine whether oral administration of B. pseudolongum affects CHS response. Viable B. pseudolongum was successfully isolated from mouse faeces. Female BALB/c mice were fed a synthetic diet with or without FOS supplementation, and B. pseudolongum (2 x 10(7) cells) was administered daily throughout the experimental period. Two weeks after starting the test diets, mice received DNFB on the ear auricle twice at 7-d intervals. Conventional cultivation and molecular biological analyses based on 16S rRNA gene sequences showed that administration of FOS and B. pseudolongum resulted in higher excretion of viable bifidobacteria, mainly B. pseudolongum. Although dietary FOS reduced the CHS response as demonstrated by ear swelling, B. pseudolongum administration resulted in a reduction in the initial phase only of the CHS response. B. pseudolongum administration increased hapten-specific IgG1, while dietary FOS decreased IgG2a in sera. Administration of FOS and B. pseudolongum decreased interferon-gamma production and increased IL-10 production in cervical lymph node cells restimulated with hapten in vitro. We conclude that B. pseudolongum proliferation in the intestinal tract is partially responsible for the reduction in DNFB-induced CHS response by dietary supplementation with FOS in mice, which may be mediated by the modulation of antigen-induced cytokine production. PMID- 20003568 TI - Tolerance of arabinoxylan-oligosaccharides and their prebiotic activity in healthy subjects: a randomised, placebo-controlled cross-over study. AB - The tolerance and prebiotic effect following oral intake by healthy human subjects of arabinoxylan-oligosaccharides (AXOS), produced by partial enzymic hydrolysis of the wheat fibre arabinoxlyan, were studied. A total of twenty healthy subjects participated in the present randomised, placebo-controlled cross over study. They consumed 10 g AXOS or placebo per d each for 3 weeks with a 4 week wash-out period in between. Before and immediately after each intake period, blood samples were taken to measure haematological and clinical chemistry parameters and the subjects completed a questionnaire about gastrointestinal symptoms. Additionally, urine was collected over 48 h for analysis of p-cresol and phenol content by GC-MS, and faeces were collected over 72 h for analysis of microbiota using real-time PCR. Of the subjects, ten also performed a urine and faeces collection 2 weeks after the start of intake (during intervention). A limited number of tested blood parameters were influenced in a statistically significantly way by either AXOS or placebo intake, but these changes remained within the normal range. Blood lipids remained unchanged. AXOS had no statistically significant effect on the range of gastrointestinal symptoms, except for a mild increase in flatulence. Urinary p-cresol excretion, an indicator of protein fermentation, was significantly decreased after 2 weeks of AXOS intake. The levels of bifidobacteria were significantly increased after 2 and 3 weeks of AXOS intake as well as after 3 weeks of placebo. However, the effect of AXOS on bifidobacteria was more pronounced than that of placebo. In conclusion, AXOS are a well-tolerated prebiotic at the dose of 10 g/d. AXOS intake increases faecal bifidobacteria and reduces urinary p-cresol excretion. PMID- 20003569 TI - Can vitamin C induce nucleotide excision repair? Support from in vitro evidence. AB - Intracellular vitamin C acts to protect cells against oxidative stress by intercepting reactive oxygen species (ROS) and minimising DNA damage. However, rapid increases in intracellular vitamin C may induce ROS with subsequent DNA damage priming DNA repair processes. Herein, we examine the potential of vitamin C and the derivative ascorbate-2-phosphate (2-AP) to induce a nucleotide excision repair (NER) response to DNA damage in a model of peripheral blood mononuclear cells. Exposure of cells to elevated levels of vitamin C induced ROS activity, resulting in increased levels of deoxycytidine glyoxal (gdC) and 8-oxo-2' deoxyguanosine (8-oxodG) adducts in DNA; a stress response was also induced by 2 AP, but was delayed in comparison to vitamin C. Evidence of gdC repair was also apparent. Measurement of cyclobutane thymine-thymine dimers (T < >T) in DNA and culture supernatant were included as a positive marker for NER activity; this was evidenced by a reduction in DNA and increases in culture supernatant levels of T < >T for vitamin C-treated cells. Genomics analysis fully supported these findings confirming that 2-AP, in particular, induced genes associated with stress response, cell cycle arrest, DNA repair and apoptosis, and additionally provided evidence for the involvement of vitamin C in the mobilisation of intracellular catalytic Fe. PMID- 20003570 TI - Effect of dietary sodium phytate and microbial phytase on the lipase activity and lipid metabolism of broiler chickens. AB - The objective of the present study was to investigate the effect of dietary phytate and microbial phytase on the lipase activity, lipid metabolism and mRNA expressions of fatty acid synthase (FASN) and leptin in broiler chickens. The study was conducted as a 2 x 3 factorial arrangement of treatments with phytate phosphorus at 0.20 and 0.40 % (added as the sodium phytate) and supplemental microbial phytase at 0, 500, or 1000 phytase units/kg. The results showed that phytase improved (P < 0.05) the growth performance and ileal digestibility of nutrients of broilers, but phytate had no effect (P>0.05) on these parameters, except the decrease (P < 0.01) in the digestibility of Ca. Phytate decreased (P < 0.05) the lipase activity, serum total cholesterol (T-CHO) and hepatic TAG, and elevated (P < 0.01) serum NEFA and HDL cholesterol. Phytase decreased (P < 0.05) serum NEFA, but increased (P < 0.01) serum T-CHO and hepatic TAG. Phytate and phytase also influenced (P < 0.01) the mRNA expressions of leptin in the liver. There were significant (P < 0.05) interactions of phytate and phytase on the concentrations of serum TAG and LDL cholesterol, hepatic NEFA and T-CHO, and the mRNA expressions of FASN. The results suggest that phytate and phytase can affect lipase activity and lipid metabolism of broiler chickens. PMID- 20003571 TI - Movement analyses of wood cricket ( Nemobius sylvestris) (Orthoptera: Gryllidae). AB - Information on the dispersal ability of invertebrate species associated with woodland habitats is severely lacking. Therefore, a study was conducted examining the movement patterns of wood cricket (Nemobius sylvestris) (Orthoptera: Gryllidae) on the Isle of Wight, UK. Juvenile (i.e. nymphs) and adult wood crickets were released and observed over time within different ground surface substrates. Their movement paths were recorded and subsequently analysed using random walk models. Nymphs were found to move more slowly than adults did; and, when given a choice, both nymphs and adults showed a preference for moving through or over leaf litter compared to bare soil or grass. A correlated random walk (CRW) model accurately described the movement pattern of adult wood crickets through leaf litter, indicating a level of directional persistence in their movements. The estimated population spread through leaf litter for adults was 17.9 cm min-1. Movements of nymphs through leaf litter could not accurately be described by a random walk model, showing a change in their movement pattern over time from directed to more random movements. The estimated population spread through leaf litter for nymphs was 10.1 cm min-1. The results indicate that wood cricket adults can be considered as more powerful dispersers than nymphs; however, further analysis of how the insects move through natural heterogeneous environments at a range of spatio-temporal scales needs to be performed to provide a complete understanding of the dispersal ability of the species. PMID- 20003572 TI - Genetic variation among Helicoverpa armigera populations as assessed by microsatellites: a cautionary tale about accurate allele scoring. AB - The existence of genetic differences among Australian populations of the pest moth Helicoverpa armigera based on microsatellite markers is contentious. To resolve this issue, we analyzed microsatellite variation in moth samples from multiple locations simultaneously in two laboratories that have previously reported contrasting patterns. Alleles and allele numbers detected in the laboratories differed, as did the genetic differences found between the samples. The automated scoring system used in one of the laboratories combined with non denaturing polyacrylamide gels led to inaccurate identification of alleles and high FST values between the populations. However, H. armigera in Australia is probably not structured geographically, with high gene flow between populations. This influences management of H. armigera and the development of area-wide control options, as populations need to be considered as one panmictic unit. The results also highlight potential problems of automated scoring systems when these are not checked carefully. PMID- 20003573 TI - Immature development and adult eclosion of Ufens principalis Owen (Hymenoptera: Trichogrammatidae), an egg parasitoid of Homalodisca spp. (Hemiptera: Cicadellidae) in southern California. AB - The biology of the immature stages and adult eclosion of Ufens principalis Owen, an important parasitoid of Homalodisca eggs in southern California, were studied. The duration of the egg, larval and pupal stages at 26.7 degrees C were 0-1, 7 and 9 days, respectively. Sacciform larvae, which developed gregariously within host eggs, were motile until about five days of age, and then became sessile. Parasitized host eggs changed from whitish and soft when freshly-laid to yellow orange and hard at five days and older. This change was accompanied by formation of septal walls separating the mature larvae and pupae. The rate of immature development had a strong positive linear relationship (R(2)=0.853, n=98) with temperatures in the range of 20.0-30.3 degrees C. The theoretical minimum threshold for immature development was 13.5 degrees C, and the required heat units were 241.0 degree-days. Adult eclosion from host eggs occurred mostly (85%) on the first two days of emergence. Although most females emerged during the morning hours (0600-1200 h), males tended to emerge earlier than females with equal emergence during the morning and late night hours (2400-0600 h). The rate of successful adult emergence was high (88%). The ratio of enclosed adults to the number of exit holes was 1.18, indicating that most adults tended to independently cut their exit holes. The number of exit holes had a strong negative relationship (R2=0.711, n=125) with exit hole size, suggesting that larger numbers of developing immatures per host egg result in an overall decrease in adult size. PMID- 20003574 TI - Determination of biotypes of Dactylopius tomentosus (Hemiptera: Dactylopiidae) and insights into the taxonomic relationships of their hosts, Cylindropuntia spp. AB - Host specialization to form biotypes is common among phytophagous insects, and it has been hypothesised that biotypes of Dactylopius tomentosus L. (Hemiptera: Dactylopiidae) occur. D. tomentosus is an important biological control agent for Cylindropuntia cacti when they occur as weeds. Additionally, there is uncertainty surrounding the taxonomic status of some species of Cylindropuntia. This study aimed to confirm the existence of D. tomentosus biotypes and to assess whether host specialization can help to resolve this systematic uncertainty. For this study, the host specificity and performance of ten provenances of D. tomentosus collected from C. cholla, C. fulgida var. fulgida, C. imbricata, C. f. var. mamillata, C. rosea and C. tunicata and reared on C. cholla, C. fulgida var. fulgida, C. imbricata and C. rosea were investigated. Five life-history parameters were measured including: crawler development time and survival, female development time, and the weight and number of eggs produced by females. Results revealed significant variation in host specificity with provenances either thriving, surviving or dying on the different hosts, thus demonstrating the existence of biotypes. Also, host specificity was related to host species and not to the geographic locality from which either the host or provenance was sourced. These findings suggest that the characteristics of Cylindropuntia species may differ sufficiently, there by presenting different selection pressures that induce and sustain distinct biotypes of D. tomentosus. The observed host use patterns of the biotypes separated the plant species into two groups that accorded with known phylogenetic relationships among Cylindropuntia species, suggesting that biotypes can be used to elucidate their taxonomic relatedness. Besides advancing our knowledge of the ecology and evolution of D. tomentosus, these novel findings have important implications for the biological control of Cylindropuntia species. PMID- 20003575 TI - Aggregation in insect communities colonizing cattle-dung. AB - Ruminant dung is a highly ephemeral, patchily distributed resource, which is used by a diverse community of invertebrate species. In such environments, high levels of insect aggregation may be important in facilitating stability and coexistence across patchily distributed populations. The aim of the present work was to quantify the aggregation of the insects colonising cow-dung in cattle pasture in southwest England and to test the hypothesis that the dung-pat community assemblage observed was the result of stochastic colonization. This was examined using batches of ten standardised, 1.5 kg, artificial cow pats placed out in cattle pastures in each of 24 weeks between May and October in 2001. Pats were left exposed in the field for seven days before being brought back to the laboratory, where any insect colonizers were collected and identified. Individual pats contained, on average, only half the number of insect taxa present in an entire batch put out at any one time. All larval coleopteran taxa, 20 of the 22 adult coleopteran taxa and 22 of the 23 dipteran taxa, showed significant aggregation, with the abundance of most taxa within pats approximating a negative binomial distribution. A simulation analysis was used to show that the observed relative frequency of taxa within pats did not differ from that expected by chance if colonisation is a random binomial event in which each species colonises a pat independently of all other species. Aggregated populations, of even highly abundant insects, may be more susceptible to the deleterious effects of insecticidal contaminants in dung than if they were evenly distributed, if by chance they colonize a pat containing insecticidal residues from a recently treated animal. PMID- 20003576 TI - Successive single-word utterances and use of conversational input: a pre syntactic route to multiword utterances. AB - In the period between sole use of single words and majority use of multiword utterances, children draw from their existing productive capability and conversational input to facilitate the eventual outcome of majority use of multiword utterances. During this period, children use word combinations that are not yet mature multiword utterances, termed 'successive single-word utterances' (SSWUs). The language development of five children, observed in play with their mothers, was studied longitudinally across the transitional period (age 1 ; 3 to 2 ; 0). Results demonstrate a common developmental trajectory from single words to SSWUs, formed with the support of conversation, to more independent SSWUs, and finally to majority use of multiword utterances. The children varied in the extent to which they produced SSWUs and whether they first produced across-turn versus within-turn SSWUs. Possible reasons for variability and why SSWU production may be important to the development of multiword utterances are discussed. PMID- 20003577 TI - Object movement in preschool children's word learning. AB - Two studies examined whether preschool children preferred to select a moving object over stationary objects when determining the referent of a novel word. In both studies three- and four-year-olds observed three novel objects, one moving object and two stationary objects. In Study 1, children (n=44) were asked to select the object that best matched a novel word. In Study 2, children (n=45) were asked to select the object that best matched a novel fact. Results across the two studies indicated that three- and four-year-olds showed a preference for selecting the moving object and that this preference was similar for both words and facts. These results suggest that preschool children are able to use movement to determine the referent of a novel word, especially when other cues are unavailable or unhelpful, but that movement may not be uniquely helpful for word learning. PMID- 20003578 TI - The effect of bovine embryo culture without proteins supplements until day 4 on transcription level of hyaluronan synthases, receptors and mtDNA content. AB - The effect of bovine embryo culture on a flat surface, (without a surface-active compound) on the level of mRNA expression of hyaluronan (HA) synthases (Has1, Has2 and Has3), Ha receptors RHAMM and C44 receptors was evaluated by mitochondrial DNA concentration and in vitro development. Cultures were evaluated up to 96 h post-insemination (hpi) using SOFaa medium. Of the three Has isoforms, Has2 expression only increased in the bovine serum albumin (BSA)-only supplemented groups regardless of time of BSA addition. Expression of RHAMM receptors was highly dependent on the addition of HA, irrespective of the presence of BSA in the medium. In contrast, expression of the CD44 receptor gene was not affected by any treatment. The cleavage rates and number of embryos that developed to 75 years and 5/6 for those aged <75 years. This study has good validity in discriminating AD participants and the data here can help in diagnosing AD and mild cognitive impairment using the CVVLT. PMID- 20003580 TI - Neuropsychological outcomes after coronary artery bypass grafting. AB - Short- and long-term cognitive declines after cardiac surgery with cardiopulmonary bypass have been reported, but the frequency, severity, nature, and etiology of postoperative cognitive changes have been difficult to quantify. Current studies have corrected the principal methodological shortcoming of earlier studies by including control groups, and have shown that while early postoperative cognitive decline does occur in some patients, it is generally mild and reversible by 3 months after surgery. Late cognitive changes do occur, but comparison with patients undergoing off-pump surgery or those being treated medically suggests that these changes are not specific to CABG or more specifically to the use of cardiopulmonary bypass. PMID- 20003581 TI - Neuropsychological performance following a history of multiple self-reported concussions: a meta-analysis. AB - Debate continues about the long-term neuropsychological impact of multiple mild traumatic brain injuries (MTBI). A meta-analysis of the relevant literature was conducted to determine the impact of having a history of more than one self reported MTBI (versus just one MTBI) across seven cognitive domains, as well as symptom complaints. The analysis was based on 8 studies, all conducted with athletes, involving 614 cases of multiple MTBI and 926 control cases of a single MTBI. The overall effect of multiple MTBI on neuropsychological functioning was minimal and not significant (d = 0.06). However, follow-up analyses revealed that multiple self-reported MTBI was associated with poorer performance on measures of delayed memory and executive functioning. The implications and limitations of these findings are discussed. PMID- 20003582 TI - Characteristics of executive function impairment in Parkinson's disease patients without dementia. AB - Executive function impairments in Parkinson's disease (PD) are well documented. However, uncertainties remain regarding the impact of these deficits on other areas of cognitive functioning. The goal of this study was to provide a comprehensive assessment of cognitive characteristics in patients with PD without dementia and to assess how any such deficits affected other areas of cognitive functioning. Forty PD patients without dementia were compared to healthy controls using measures of attention and speed of processing and a comprehensive set of executive function tests including working memory, planning, and problem solving. Measures of memory/learning and visuospatial skills were also included to examine the relationship between aspects of executive function and other areas of cognition. Patients with PD showed deficits on measures of executive function, problem solving, and visuospatial skills. However, they were unimpaired on measures of planning, attention, and memory/learning. Deficits in problem solving were only evident for tasks with a high visuospatial content and were no longer significant when visuospatial skills were controlled for. While deficits in executive function and visuospatial skills were apparent for PD patients compared to controls, many aspects of cognition remained intact. These can provide a focus for cognitive intervention strategies that can be effective in delaying decline for PD patients. PMID- 20003583 TI - Interstimulus jitter facilitates response control in children with ADHD. AB - Interstimulus "jitter" involves randomization of intervals between successive stimulus events, and can facilitate performance on go/no-go tests among healthy adults, though its effect in clinical populations is unclear. Children with Attention-deficit/Hyperactivity Disorder (ADHD) commonly exhibit deficient response control, leading to increased intra-subject variability (ISV), which has been linked to anomalous functioning within frontal circuits, as well as their interaction with posterior "default mode" regions. We examined effects of interstimulus jitter on response variability in 39 children, ages 9-14 years (25 ADHD, 14 controls). Participants completed 2 computerized go/no-go tests: one with fixed interstimulus interval (ISI) and one with jittered ISI. Repeated measures analysis of variance (ANOVA) revealed a significant group-by test interaction, such that introduction of jitter produced a significant decrease in ISV among children with ADHD, but not among controls. Whereas children with ADHD were significantly more variable than controls on the go/no-go test with fixed ISI, their performance with jittered ISI was equivalent to that of controls. Jittering stimulus presentation provides a nonpharmacologic mechanism for improving response control in ADHD. This bottom-up approach may be mediated by increases in vigilance through noradrenergic circuits that facilitate maintenance of frontal circuits critical to response control. PMID- 20003584 TI - Longitudinal patterns of semantic and episodic memory in frontotemporal lobar degeneration and Alzheimer's disease. AB - The longitudinal assessment of episodic and semantic memory was obtained from 236 patients diagnosed with Alzheimer's disease (AD, n = 128) and with frontotemporal lobar degeneration (FTLD, n = 108), including patients with a social comportment/dysexecutive (SOC/EXEC) disorder, progressive nonfluent aphasia (PNFA), semantic dementia (SemD), and corticobasal syndrome (CBS). At the initial assessment, AD patients obtained a lower score on the delayed free recall test than other patients. Longitudinal analyses for delayed free recall found converging performance, with all patients reaching the same level of impairment as AD patients. On the initial evaluation for delayed recognition, AD patients also obtained lower scores than other groups. Longitudinal analyses for delayed recognition test performance found that AD patients consistently produced lower scores than other groups and no convergence between AD and other dementia groups was seen. For semantic memory, there were no initial between-group differences. However, longitudinal analyses for semantic memory revealed group differences over illness duration, with worse performance for SemD versus AD, PNFA, SOC/EXEC, and CBS patients. These data suggest the presence of specific longitudinal patterns of impairment for episodic and semantic memory in AD and FTLD patients suggesting that all forms of dementia do not necessarily converge into a single phenotype. PMID- 20003585 TI - Topical 5-fluorouracil for granular myringitis: a double-blinded study. AB - BACKGROUND: Granular myringitis is a poorly understood disease which causes considerable discomfort and concern to affected individuals. Some cases may progress to inflammatory obliteration of the deep ear canal. Treatment options are diverse, and no single accepted treatment modality exists. Local application of 5-fluorouracil has recently been trialled in the treatment of chronic otitis media, with promising results. AIM: To investigate the efficacy and safety of topical 5-fluorouracil in cases of granular myringitis. METHODS: Sixty patients with granular myringitis were enrolled in this controlled, double-blinded study. Patients were divided randomly into two equal groups. Patients in the study group (n = 30) received three successive applications of topical 5-fluorouracil 5 per cent cream, with two-week intervals between treatments. Patients in the control group (n = 30) received only local petroleum jelly cream packing at the same intervals. Follow-up appointments were scheduled every two weeks for the first three months; patients were then seen again after two years for evaluation of any persistent disease. Outcome measures were: (1) after three months, symptomatic response to treatment (i.e. quantity of aural discharge as reported by patient) and change in the amount of granulation tissue (as assessed by the physician via endoscopic examination); and (2) persistent disease after two years. RESULTS: There was a highly statistically significant difference in the studied outcomes, comparing the study and control groups. CONCLUSION: Local application of 5 fluorouracil is a new treatment method for granular myringitis. In this study, it proved safe and efficacious, with no serious local reactions or complications. PMID- 20003586 TI - Ectopic cervical thymus: case report and review of pathogenesis and management. AB - OBJECTIVE: Ectopic cervical thymus is a rare anomaly, and 50 per cent of cases occur in children. The aim of this report was to remind clinicians of this rare condition, which is part of the differential diagnosis of a paediatric neck lump. METHOD: Case report together with literature review of the pathogenesis and management of ectopic cervical thymus. RESULTS: The ectopic cervical thymus is a rare cause of a benign neck mass, and is thus usually misdiagnosed as lymphadenopathy or neoplasia. We discuss its embryology, aetiology, presentation, histopathology, radiology and management, with reference to the reported case. CONCLUSION: Ectopic cervical thymus should always be considered in the differential diagnosis of a paediatric neck lump. The diagnosis can often be confirmed by cytology and radiology prior to surgical excision. PMID- 20003587 TI - Functional status after total laryngectomy: cross-sectional survey of 79 laryngectomees using the Performance Status Scale for Head and Neck Cancer. AB - INTRODUCTION: Total laryngectomy affects patients' post-operative speech and swallowing functions. We aimed to assess these outcomes. MATERIALS AND METHODS: Patients' normalcy of diet, ability to eat in public and speech comprehensibility were assessed using the Performance Status Scale for Head and Neck Cancer, in a cross-sectional survey of disease-free laryngectomees. RESULTS AND ANALYSIS: Seventy-nine laryngectomees (72 men and seven women), with a mean age of 64 years (range 37 to 96), were included. Mean scores were 81.6 (standard deviation 29.2) for normalcy of diet, 77.8 (standard deviation 30.2) for eating in public and 65.2 (standard deviation 23.5) for speech. Normalcy of diet achieved higher scores within six months of laryngectomy and remained stable. There was a statistically significant difference between scores for the speech and public eating domains, comparing patients less and more than six months post laryngectomy. This trend was maintained beyond 12 months. CONCLUSIONS: In post laryngectomy patients, Performance Status Scale for Head and Neck Cancer scores improved over time, especially those for the speech and public eating domains, reflecting increasing confidence in social interactions and familiarity with surgical voice restoration. PMID- 20003588 TI - Editorial. PMID- 20003589 TI - The effect and mode of action of saponins on the microbial populations and fermentation in the rumen and ruminant production. AB - The growing public concerns over chemical residues in animal-derived foods and threats of antibiotic-resistant bacteria have renewed interest in exploring safer alternatives to chemical feed additives in ruminant livestock. Various bioactive phytochemicals including saponins appear to be potential 'natural' alternatives to 'chemical' additives in modulating rumen fermentation favourably and animal performance. Saponins are a diverse group of glycosides present in many families of plants. The primary effect of saponins in the rumen appears to be to inhibit the protozoa (defaunation), which might increase the efficiency of microbial protein synthesis and protein flow to the duodenum. Furthermore, saponins may decrease methane production via defaunation and/or directly by decreasing the activities (i.e. rate of methanogenesis or expression of methane-producing genes) and numbers of methanogens. Saponins may also selectively affect specific rumen bacteria and fungi, which may alter the rumen metabolism beneficially or adversely. The ammonia-adsorption and modulation of digesta passage in the rumen by saponins have also been implicated in altering rumen metabolism, but their physiological responses are likely to be negligible compared with microbiological effects. The effects of saponins on rumen fermentation have not been found to be consistent. These discrepancies appear to be related to the chemical structure and dosage of saponins, diet composition, microbial community and adaptation of microbiota to saponins. There is need for systematic research based on chemical structures of saponins, nutrient composition of diets and their effects on rumen microbial ecosystem to obtain consistent results. The present paper reviews and discusses the effects and mode of action of saponins on microbial community and fermentation in the rumen, and ruminant performance. PMID- 20003590 TI - Functional foods for dyslipidaemia and cardiovascular risk prevention. AB - A food can be regarded as 'functional' if it can demonstrate a beneficial efficacy on one or more target functions in the body in a convincing way. Beyond adequate nutritional qualities, functional foods should either improve the state of health and wellbeing and/or reduce the risk of disease. Functional foods that are marketed with claims of heart disease reduction focus primarily on the major risk factors, i.e. cholesterol, diabetes and hypertension. Some of the most innovative products are designed to be enriched with 'protective' ingredients, believed to reduce risk. They may contain, for example, soluble fibre (from oat and psyllium), useful both for lowering cholesterol and blood pressure, or fructans, effective in diabetes. Phytosterols and stanols lower LDL-cholesterol in a dose-dependent manner. Soya protein is more hypocholesterolaemic in subjects with very high initial cholesterol and recent data indicate also favourable activities in the metabolic syndrome. n-3 Fatty acids appear to exert significant hypotriacylglycerolaemic effects, possibly partly responsible for their preventive activity. Dark chocolate is gaining much attention for its multifunctional activities, useful both for the prevention of dyslipidaemia as well as hypertension. Finally, consensus opinions about tea and coffee have not emerged yet, and the benefits of vitamin E, garlic, fenugreek and policosanols in the management of dyslipidaemia and prevention of arterial disease are still controversial. PMID- 20003591 TI - Congenital cholesteatoma of mastoid region manifesting as acute mastoiditis: case report and literature review. AB - OBJECTIVES: We report an extremely rare case of congenital cholesteatoma of the mastoid region, presenting as acute mastoiditis. We also review the 16 previously reported cases of congenital cholesteatoma of the mastoid region. CASE REPORT: A 65-year-old man presented with left-sided, post-auricular swelling and pain. Acute mastoiditis was diagnosed, with computed tomography demonstrating destruction of the bony plates of the posterior cranial fossa and sigmoid sinus. Initial surgery revealed a cholesteatoma in the mastoid, with no extension into the aditus ad antrum or attic. These findings were confirmed by pathological and immunohistochemical analysis of the surgical specimen, the latter using involucrin. The cholesteatoma matrix was completely removed in a second operation. CONCLUSIONS: Including this case, only four of the 17 reported cases of congenital cholesteatoma of the mastoid region showed post-auricular pain or swelling, indicating acute mastoiditis. Clinicians should bear in mind that congenital cholesteatoma may be present in patients presenting with mastoiditis, particularly adults. PMID- 20003592 TI - A non-traumatic, blue-purple auricle: case report. AB - OBJECTIVE: We report a typical case of earlobe lymphocytoma. METHOD: A case report and literature review are presented. RESULTS: A 10-year-old girl presented with a blue-coloured earlobe. A diagnosis of Lyme disease was confirmed by serological tests. Lyme borreliosis is the most common tick-borne disease in the northern hemisphere. It is caused by the spirochete Borrelia burgdorferi sensu lato. The patient was successfully treated with antibiotics. CONCLUSION: The diagnostic process and ENT symptomatology of Lyme disease and borrelial lymphocytoma are summarised and discussed. PMID- 20003593 TI - Stridor in Parkinson's disease: a case of 'dry drowning'? AB - OBJECTIVES: (1) To present a rare case of stridor secondary to prolonged laryngospasm in a patient with Parkinson's disease, and (2) to review the literature on stridor in Parkinson's disease. METHODS: We report a 73-year-old Parkinson's disease patient who developed acute stridor due to prolonged laryngospasm triggered by overspill of excessive secretions. The literature was reviewed, following a Medline search using the keywords 'Parkinson's disease' and 'stridor' or 'airway obstruction' or 'laryngospasm' or 'laryngeal dystonia' or 'bilateral vocal cord palsy'. RESULT: Only 12 previously reported cases of stridor in Parkinson's disease patients were identified. Causes included bilateral vocal fold palsy (eight cases), laryngospasm (five), and dystonia of the jaw and neck muscles (two). The mechanism of laryngospasm in our patient was similar to 'dry drowning', and has not previously been described. CONCLUSION: Laryngospasm can be triggered in Parkinson's disease by excessive secretions entering the larynx. The mechanism is similar to 'dry drowning'. Treatment focuses on reducing secretions. The use of botulinum toxin to reduce spasm is inappropriate in this situation. This case emphasises the importance of recognising different causes of stridor in Parkinson's disease patients, as this affects management. PMID- 20003594 TI - Inferior vestibular neuritis: a novel subtype of vestibular neuritis. AB - OBJECTIVE: To report eight cases of inferior vestibular neuritis, in order to raise awareness of this new subtype of vestibular neuritis. MATERIALS AND METHODS: We retrospectively analysed 216 patients (104 males and 112 females; age range 10-64 years; mean age 38.4 years) with full clinical documentation who had attended our hospital's vertigo clinic between May 2007 and December 2008. All patients underwent systematic investigation, including hearing tests, radiology, caloric testing and vestibular evoked myogenic potential testing. RESULTS: Of 216 patients with vestibular neuritis, eight cases were diagnosed as inferior vestibular neuritis, based on comprehensive analysis of test data. The clinical features of these eight patients were consistent with the characteristics of vestibular neuritis. The results of pure tone audiometry and caloric testing were normal, and the possibility of central lesions was excluded by cerebral computed tomography or magnetic resonance imaging on admission. Six cases had unilateral loss of vestibular evoked myogenic potentials, whereas two had a unilateral lower amplitude of vestibular evoked myogenic potentials. CONCLUSIONS: Inferior vestibular neuritis is a novel subtype of vestibular neuritis, which involves the inferior vestibular nerve alone. Vestibular evoked myogenic potential testing is a useful aid to the diagnosis of inferior vestibular neuritis. PMID- 20003595 TI - Radiofrequency coblation for treatment of advanced laryngotracheal recurrent respiratory papillomatosis. AB - BACKGROUND: A variety of treatment modalities are currently used to treat recurrent respiratory papillomatosis. We aimed to study the efficacy of radiofrequency cold ablation (coblation) for the treatment of laryngotracheal recurrent respiratory papillomatosis, by comparing treatment intervals for coblation and CO2 laser vaporisation. METHOD: Retrospective case series of adult patients with advanced laryngotracheal recurrent respiratory papillomatosis. RESULTS: Six patients were treated for at least two years by CO2 laser vaporisation with or without intralesional cidofovir. All six subsequently underwent treatment with radiofrequency coblation with or without intralesional cidofovir. Coblation resulted in longer periods between interventions, compared with CO2 laser (p = 0.03). CONCLUSION: Radiofrequency coblation appears to be an attractive alternative technique to CO2 laser for the surgical treatment of advanced laryngotracheal papillomata. PMID- 20003596 TI - Neuro-otological findings in tinnitus patients with normal hearing. AB - INTRODUCTION: Tinnitus is usually associated with hearing loss, and patients with tinnitus and normal hearing are unusual. Neuro-otological findings have not previously been described in tinnitus patients with normal hearing. AIM: To analyse neuro-otological examination results from a group of tinnitus patients with normal hearing. MATERIALS AND METHODS: Seventeen normal-hearing tinnitus patients seen over a 10-year period were retrospectively evaluated. Their results were compared with those of a control group of 17 normal subjects without tinnitus. RESULTS: The main neuro-otological finding in the tinnitus patients was caloric test abnormality: a unilateral canal paresis was present in 15 of the 17 patients. Caloric tests were normal in 15 of the 17 control subjects. CONCLUSION: We may infer from these results that tinnitus could be the only clinical manifestation of a cochlear - and presumably cochleo-vestibular - lesion, and that unilateral canal paresis may be the only abnormal finding on neuro otological examination. PMID- 20003597 TI - Plasticity of fixation in patients with central vision loss. AB - The aim of this study was to explore the plasticity of fixation in patients with central vision loss. Most of these patients use preferred retinal loci (PRLs) in the healthy eccentric part of the retina to fixate, but fixation stability and retinal location are not always optimal for best visual performance. This study examined whether fixation stability and a new PRL location can be trained and whether these changes in ocular motor control transfer into better reading performance. Six patients with age-related macular degeneration participated in the study. Fixation stability measurements, microperimetry, and auditory biofeedback training were performed with the MP-1 microperimeter. The auditory biofeedback was used during five 1-h long training sessions to improve fixation and relocate the PRL. Fixation location and stability were recorded while viewing four different targets: a cross, a letter, a word, and a nine-cycle radial grating. Visual acuity was assessed with the Early Treatment Diabetic Retinopathy Study (ETDRS) chart and reading performance with the MNRead test. The results showed that all patients developed a new PRL in an optimal location for reading, and they were able to use it consistently while viewing different targets. Fixation stability improved 53% after training. Learning transferred to the old PRL even though fixation stability at this location was not trained. All these improvements in ocular motor control translated into better reading performance: reading speed improved 38% and reading acuity and critical print size gained two lines. We conclude that the ability of the ocular motor system to fixate is flexible in patients with central vision loss: a new PRL can be trained, fixation stability can be improved, and learning transfers to an untrained location. These gains in ocular motor control result in better visual performance. This property can be successfully used to optimize the residual vision of patients with central vision loss. PMID- 20003598 TI - Cell type-specific and light-dependent expression of Rab1 and Rab6 GTPases in mammalian retinas. AB - The Ras-like Rab1 and Rab6 GTPases modulate protein traffic along the early secretory pathway and are involved in the regulation of maturation of rhodopsin in the outer retina. However, Rab GTPases have not been studied in the inner retinas. Here, we analyzed the anatomatic distribution and expression of Rab1 and Rab6 in the mouse and rat retinas by immunohistochemistry and immunoblotting. We found that Rab1 was specifically expressed in the rod bipolar cells, while Rab6 was expressed in a different cell type(s) from rod bipolar cells in the inner retina. We also demonstrated that expression of Rab1 and Rab6 was increased with light. These data provided the first evidence implicating that Rab1 and Rab6 may be involved in the regulation of the retinal adaptation. PMID- 20003599 TI - Endoscopic, endonasal decompression of spinal stenosis with myelopathy secondary to cranio-vertebral tuberculosis: two cases. AB - BACKGROUND: Tuberculosis can cause extensive osseo-ligamentous destruction at the cranio-vertebral junction, leading to atlanto-axial instability and compression of vital cervico-medullary centres. This may manifest as quadriparesis, bulbar dysfunction and respiratory insufficiency. AIM: We report two patients presenting with spinal stenosis and cord compression secondary to cranio-vertebral tuberculosis, who were successfully decompressed via an endoscopic, endonasal approach. STUDY DESIGN: Two case reports. METHODS AND RESULTS: Both patients were successfully decompressed via an endoscopic, endonasal approach which provided access to the cranio-vertebral junction and upper cervical spine. CONCLUSION: An endoscopic, endonasal approach is feasible for the surgical management of cranio vertebral junction stenosis; such an approach minimises surgical trauma to critical structures, reducing post-operative morbidity and the duration of hospital stay. PMID- 20003600 TI - Common carotid artery pseudoaneurysm formation following foreign body ingestion. AB - OBJECTIVE: To report an unusual case of pseudoaneurysm formation following ingestion of a thin wire. METHOD: Case report, including management, and review of the world literature concerning pseudoaneurysm and its management. RESULTS: A 15-year-old boy presented as an emergency with a two-week history of painful swallowing, and a one-week history of a progressively enlarging, right-sided swelling of the lower neck. A foreign body had been ingested two weeks previously. Radiological investigation showed a pseudoaneurysm of the right common carotid artery. The neck was explored, the foreign body removed and the common carotid artery repaired. The patient was discharged on the 10th post operative day without neurovascular complication. CONCLUSION: This is an unusual presentation of pseudoaneurysm of the common carotid artery following accidental ingestion of a foreign body, reported for its rarity and management. PMID- 20003601 TI - Does antrochoanal polyp present with epistaxis? AB - OBJECTIVE: To compare the gross and microscopic appearance of antrochoanal polyps associated with recurrent epistaxis, with those with a more typical presentation. DESIGN: Prospective, controlled study. METHODS: All patients underwent clinical and endoscopic examination, computed tomography scanning, and examination under anaesthesia, in order to detect the gross diagnostic criteria for antrochoanal polyp. Histological findings on light microscopy were compared for polyps presenting with epistaxis versus those without. The number of predominant inflammatory cells in the corium was determined in both groups and statistically compared using the Student t-test. RESULTS: Recurrent epistaxis was a presenting symptom in 10/84 (11.9 per cent) patients with gross diagnostic criteria for antrochoanal polyp. Grossly, these patients' polyps had a reddish, vascular surface in parts. Histologically, these polyps showed a highly vascular stroma with multiple dilated blood vessels, the typical appearance of an angiomatous antrochoanal polyp. Thrombi at different stages of development were detected, with no infarcts. The remaining cases (88.1 per cent) had no history of epistaxis; histologically, these patients' polyps showed an oedematous connective tissue core with few inflammatory cells. Plasma cells were predominant in the angiomatous polyps, being significantly more prevalent than in the ordinary antrochoanal polyps (p < 0.00). CONCLUSIONS: It would appear that only angiomatous antrochoanal polyps present with epistaxis. Detection of the characteristic gross appearance of these polyps may help avoid unwanted surgery. Histopathological analysis confirms the diagnosis. A significantly increased number of plasma cells may be the underlying cause of the histological changes seen in angiomatous antrochoanal polyps. PMID- 20003602 TI - Oncogenic osteomalacia in a patient with an ethmoid sinus tumour. AB - OBJECTIVE: To highlight the clinical presentation and management of a rare case of oncogenic osteomalacia due to an ethmoid sinus tumour. MATERIALS AND METHODS: We examined the case records of a 55-year-old man who presented with progressive fatigue, weakness and bone pain, and noted the clinical presentation, laboratory investigations, computed tomography findings, operative notes and follow-up details. CONCLUSION: Oncogenic osteomalacia secondary to a paranasal sinus neoplasm is a rare entity. The causative tumour is often occult and may be missed by routine clinical examination. This case report illustrates the appropriate pattern of evaluation and management to ensure a successful outcome. PMID- 20003603 TI - Outcome of computer-assisted surgery in patients with chronic rhinosinusitis. AB - OBJECTIVE: To compare the complication rates and outcome of computer-assisted versus non-computer-assisted functional endoscopic sinus surgery. METHODS: We reviewed retrospectively the medical records of 276 patients who had undergone sinus surgery for chronic rhinosinusitis with (n = 108) or without (n = 168) computer assistance, from 1996 to 2004, to determine the incidence of complications and need for revision surgery. RESULTS: The incidence of complications was 6.5 per cent in the computer-assisted group and 6.0 per cent in the non-computer-assisted group (p = 1.00). In the computer-assisted group, 9.2 per cent needed revision surgery, compared with 10.7 per cent in the non-assisted group (p = 0.84). CONCLUSIONS: Although our study found no significant difference in complications or revision rates, computer-assisted surgery serves as an important orientation aid during functional endoscopic sinus surgery. PMID- 20003604 TI - Closed rhinoplasty approach for excision of nasal dermoids. AB - OBJECTIVE: To assess the efficacy of excision of nasal dermoids through a closed rhinoplasty incision. This is the first description of the use of this approach for excision of superficial nasal dermoids. METHODS: Three boys aged five, nine and 12 years presented with midline nasal dermoids with minimal cutaneous involvement. Magnetic resonance imaging demonstrated distinct, cystic, superficial nasal masses. The cysts were excised through a closed rhinoplasty approach. In each case, completeness of extirpation was judged by histopathological examination of the excised specimen. Aesthetic outcome was recorded photographically. RESULTS: All three patients' cysts were completely excised, with excellent cosmetic results. CONCLUSIONS: The closed rhinoplasty incision is another approach in the surgeon's armamentarium for excision of small, superficial nasal dermoid cysts. In well selected cases, this approach gives optimal cosmetic results, provides adequate exposure with minimal dissection, and allows total extirpation. PMID- 20003605 TI - Self-limiting benign paroxysmal positional vertigo following use of whole-body vibration training plate. AB - OBJECTIVE: We describe a case of benign paroxysmal positional vertigo which occurred after use of a whole-body vibration training plate. METHOD: Case report and literature review concerning the secondary causes of benign paroxysmal positional vertigo and the physiological effects of whole-body vibration training plates. RESULTS: A 44-year-old woman was referred with classic symptoms of benign paroxysmal positional vertigo following use of a whole-body vibration training plate, a popular form of fitness equipment widely used in sports, rehabilitation and beauty treatments. The condition resolved spontaneously after several days. There have been reports of negative side effects in users of this equipment, such as dizziness, headache and a sensation of imbalance; however, there have been no reported cases involving vertigo. Based on a literature review, this equipment may cause side effects, including vertigo, by generating forces that can increase the original amplitude of internal organs, which may potentially cause labyrinthine trauma or dislocation of otoconia, leading to benign paroxysmal positional vertigo. CONCLUSIONS: We suggest that whole-body vibration training plates may potentially induce benign paroxysmal positional vertigo. Manufacturers may need to make users of this equipment aware of this risk, and remind them to use it with caution. PMID- 20003606 TI - Vestibular schwannoma: role of conservative management. AB - OBJECTIVE: To assess the outcome of conservative management of vestibular schwannoma. STUDY DESIGN: Observational study. SETTING: Tertiary referral centre. PATIENTS: Four hundred and thirty-six patients with vestibular schwannoma (490 tumours), including 327 sporadic tumours and 163 tumours in 109 patients with neurofibromatosis type two. MAIN OUTCOME MEASURES: The relationship of tumour growth to tumour size at presentation, and to certain demographic features. RESULTS: The initial tumour size was significantly larger in the neurofibromatosis type two group (11 mm) than in the sporadic vestibular schwannoma group (5.1 mm). In both groups, 68 per cent of tumours did not grow during follow up (mean 3.6 years; range one to 14 years). The mean growth rate was 1.1 mm/year (range 0-15 mm/year) for sporadic tumours and 1.7 mm/year (range 0-18 mm/year) for neurofibromatosis type two tumours. The tumour growth rate correlated positively with tumour size in the sporadic tumour group, and correlated negatively with age in the neurofibromatosis type two group. CONCLUSION: Two-thirds of vestibular schwannomas did not grow. Radiological surveillance is an acceptable approach in carefully selected patients. Once a sporadic vestibular schwannoma reaches 2 cm in intracranial diameter, it is likely to continue growing. We do not recommend conservative management for sporadic tumours with an intracranial diameter of 1.5 cm or more. Vestibular schwannoma management is more complex in patients with neurofibromatosis type two. PMID- 20003607 TI - Molecular pathophysiology of psoriasis and molecular targets of antipsoriatic therapy. AB - Psoriasis is a chronic inflammatory skin disease characterised by elevated red scaly plaques on specific body sites. Histologically, the plaques are defined by epidermal hyperplasia, epidermal and dermal infiltration by leukocytes, and changes in the dermal microvasculature. Differentiation and activation are disturbed in lesional psoriatic keratinocytes, and the pool of proliferating keratinocytes is increased, which is accompanied by enhanced production of proinflammatory cytokines, adhesion molecules and antimicrobial peptides. These changes in psoriatic keratinocytes are caused by altered expression of genes associated with epidermal differentiation, and by activation of signalling pathways involving signal transducer and activator of transcription 3 (STAT3), type I interferon (IFN) and mitogen-activated protein kinase (MAPK). The number of T cells, and myeloid and plasmacytoid dendritic cells (DCs) is markedly increased in psoriatic lesions. Myeloid DCs produce interleukin (IL)-23, tumour necrosis factor (TNF)-alpha and inducible nitric oxide synthase (iNOS), which are crucial cytokines in the pathogenesis of psoriasis. IL-23 stimulates the secretion of IL-22 by T helper 17 cells, and IL-22 induces epidermal hyperplasia. The crosstalk between keratinocytes and leukocytes via their proinflammatory cytokines creates the vicious circle of chronic skin inflammation seen in psoriasis. This suggests that optimal treatment of psoriasis needs to target pathogenic pathways in both leukocytes and keratinocytes. PMID- 20003608 TI - The bionomics of an invasive species Sitona lepidus during its establishment in New Zealand. AB - The egg, larval, pupal and adult abundance of the clover root weevil Sitona lepidus Gyllenhal (Coleoptera: Curculionidae) was monitored at three sites for the first ten years following the discovery of this exotic pest in the Waikato region of New Zealand. The species went through an initial boom and bust cycle at two sites, with populations reaching up to 1800 larvae m-2. Thereafter, winter larval populations were relatively stable, ranging between 450-750 m-2. Unlike in the Northern Hemisphere, S. lepidus was found to have two generations a year in the Waikato region of New Zealand. Pasture white clover content at the time of peak adult numbers was positively related to the subsequent peak larval populations for each generation. The factors contributing to the emergence of S. lepidus as one of the most important pasture pests in New Zealand are discussed. PMID- 20003609 TI - The effect of fish meal replacement by soyabean products on fish growth: a meta analysis. AB - Meta-analysis was applied to quantify the effect of replacement of fish meal by soyabean products in diets on fish growth. Measurement of growth in different units among studies required the use of a standardised effect size (Hedges' d). From a total of ninety-nine studies concentrating on fish meal replacement by defatted soyabean meal, 53 % were eliminated due to, among others, absence of a fish meal control diet (n 18), or no statistical differences or measurement of dispersion (n 6) indicated. Replacement of 4 to 40 % fish meal by soyabean meal (inclusion levels of 71-366 g/kg) resulted in a mean effect size of - 0.1142 (95 % CI - 0.4665, 0.2382) obtained in forty-eight comparisons evaluated with seventeen different fish species. However, at higher fish meal replacement levels the 95 % CI calculated for combined effect sizes did not overlap with zero. With soya protein concentrate replacing 25 to 100 % of fish meal in diets for seven fish species, methionine supplementation (mean - 2.4373 (95 % CI - 3.9004, - 0.9742); n 10) did not have a substantial influence on the magnitude of cumulative effect sizes relative to no supplementation (mean - 2.7306 (95 % CI - 3.7991, - 1.6620); n 16). Information on other soyabean products (full-fat soyabeans, soya flour) used as protein sources in fish diets was found as too limited for analysis and definite conclusions. The present study contributes by putting a numerical value to the magnitude of growth differences in fish when replacing dietary fish meal by soyabean products. PMID- 20003610 TI - Cognitive and home environmental predictors of change in sugar-sweetened beverage consumption among adolescents. AB - Sugar-sweetened beverage (SSB) consumption may increase risk for unnecessary weight gain. To develop interventions discouraging consumption, more insight is needed about cognitive and environmental predictors related to the decrease in SSB consumption. The present paper aims (1) to describe the relationship between potential cognitive determinants of change (attitudes, subjective norms, perceived behavioural control and intentions) and perceived environmental factors (family food rule and home availability of SSB) with changes in SSB consumption between baseline and 4-month follow-up and (2) to study whether the relationships between the environmental factors and SSB consumption are mediated by the cognitive determinants. Information on possible predictors and SSB intake at baseline and 4-month follow-up was provided by 348 Dutch adolescents (aged 12-13 years) through online questionnaires that were completed at school. Multilevel logistic regression and mediation analyses were used to determine direct and indirect associations between predictors and behaviour. The present results show that a high perceived behavioural control to decrease intake at baseline was associated with a decrease in consumption of SSB between baseline and follow-up (OR = 0.53). Low availability and a stricter family food rule were associated with a decrease in SSB consumption between baseline and follow-up (OR = 2.39, 0.54). The association between availability and decrease in SSB consumption was for 68 % mediated by perceived behavioural control to drink less. In conclusion, interventions to decrease SSB intake should focus on improving attitudes and perceived behavioural control to reduce intake, and on limiting home availability and stimulating stricter family food rules regarding SSB consumption. PMID- 20003611 TI - Dietary glycaemic index, dietary glycaemic load and incidence of myocardial infarction in women. AB - The association of dietary glycaemic index (GI) and glycaemic load (GL) with CVD has been examined in several populations with varying results. We tested the hypothesis that women with diets high in GI or GL would have higher rates of myocardial infarction (MI), and the associations would be stronger in overweight women. We measured dietary GI and GL in 36,234 Swedish Mammography Cohort participants aged 48-83 years using FFQ. Cox models were used to calculate incidence rate ratios (RR) and 95 % CI for hospitalisation or death due to MI assessed using the Swedish inpatient and cause-of-death registers from 1 January 1998 until 31 December 2006. Over 9 years of follow-up, 1138 women were hospitalised or died due to a first MI. In multivariable-adjusted models, the RR comparing top to bottom quartile of dietary GI were 1.12 (95 % CI 0.92, 1.35, P trend = 0.24), and the RR comparing top to bottom quartile of dietary GL were 1.22 (95 % CI 0.90, 1.65, P-trend = 0.23). Among overweight women, the RR comparing top to bottom quartile of dietary GI were 1.20 (95 % CI 0.91, 1.58, P trend = 0.22), and the RR comparing top to bottom quartile of dietary GL were 1.45 (95 % CI 0.93, 2.25, P-trend = 0.16). There were no statistically significant associations of dietary GI or GL with MI in this population. PMID- 20003613 TI - Variation in the host ABO blood group may be associated with susceptibility to hepatitis C virus infection. AB - This study aimed to determine the relationship between hepatitis C virus (HCV) infection and ABO/Rhesus blood groups, age and sex. A total of 20 000 patients who came to donate blood in the blood bank of GSVM Medical College, Kanpur were enrolled in the study. Demographic data recorded for each patient included age, sex and blood group. Blood samples were tested for anti-HCV antibodies and ABO/Rhesus blood group antigen typing was performed. The overall positive rate of anti-HCV was 0.34%. We found that seropositivity for HCV increased with age. Anti HCV antibodies were detected in 1/765 women (0.13%), compared to 67/19 235 men (0.35%). Seroprevalence of HCV was found to be higher in blood group O individuals (0.42%) and lowest in blood group AB individuals (0.04%). The results of this study demonstrate that that HCV infection may not be related to age and sex but the possible association of blood group antigens with HCV infection cannot be ruled out. PMID- 20003612 TI - Undernutrition, fatty acid and micronutrient status in relation to cognitive performance in Indian school children: a cross-sectional study. AB - While undernutrition and anaemia have previously been linked to poor development of children, relatively little is known about the role of B-vitamins and fatty acids on cognition. The present study aims to explore the associations between indicators of body size, fatty acid and micronutrient status on cognitive performance in 598 Indian school children aged 6-10 years. Baseline data of a clinical study were used to assess these associations by analyses of variance adjusting for age, sex, school, maternal education and cognitive tester. The Kaufman Assessment Battery for Children II was used to measure four cognitive domains, including fluid reasoning, short-term memory, retrieval ability and cognitive speediness. Scores were combined into an overall measure, named mental processing index (MPI). Body size indicators and Hb concentrations were significantly positively related to cognitive domains and MPI, such that increases of 1 sd in height-for-age and weight-for-age z-scores would each translate into a 0.09 sd increase in MPI, P = 0.0006 and 0.002, respectively. A 10 g/l increase in Hb concentrations would translate into a 0.08 sd increase in MPI, P = 0.0008. Log-transformed vitamin B12 concentrations were significantly inversely associated with short-term memory, retrieval ability and MPI (beta (95 % CI) = - 0.124 (- 0.224, - 0.023), P = 0.02). Other indicators of Fe, iodine, folate and fatty acid status were not significantly related to cognition. Our findings for body size, fatty acids and micronutrients were in agreement with previous observational studies. The inverse association of vitamin B12 with mental development was unexpected and needed further study. PMID- 20003614 TI - 'Feral' and 'wild'-type methicillin-resistant Staphylococcus aureus in the United Kingdom. AB - Circulation of methicillin-resistant Staphylococcus aureus (MRSA) outside hospitals could alter the impact of hospital-based control strategies. We investigated two groups of cases (each matched to controls with MRSA): 61 'community cases' not in acute hospital in the year before MRSA isolation; and 21 cases with ciprofloxacin-sensitive (CipS) MRSA. Multi-locus sequence typing, spa typing and Panton-Valentine leukocidin gene testing were performed and demographics obtained. Additional questionnaires were completed by community case GPs. Community cases comprised 6% of Oxfordshire MRSA. Three community cases had received no regular healthcare or antibiotics: one was infected with CipS. Ninety one percent of community cases had healthcare-associated sequence type (ST)22/36; CipS MRSA cases had heterogeneous STs but many had recent healthcare exposure. A substantial minority of UK MRSA transmission may occur outside hospitals. Hospital strains are becoming 'feral' or persisting in long-term carriers in the community with regular healthcare contacts; those with recent healthcare exposure may nevertheless acquire non-hospital epidemic MRSA strains in the community. PMID- 20003615 TI - Molecular characterization of SAT 2 foot-and-mouth disease virus from post outbreak slaughtered animals: implications for disease control in Uganda. AB - In Uganda, limiting the extent of foot-and-mouth disease (FMD) spread during outbreaks involves short-term measures such as ring vaccination and restrictions of the movement of livestock and their products to and from the affected areas. In this study, the presence of FMD virus RNA was investigated in cattle samples 3 months after FMD quarantine measures had been lifted following an outbreak in 2004. Oropharyngeal tissue samples were obtained from 12 cattle slaughtered in a small town abattoir in Kiboga. FMD virus RNA was detected by diagnostic RT-PCR in nine of the 12 tissue samples. Part of the coding region for the capsid protein VP1 was amplified and sequenced. All samples were identified as belonging to the SAT 2 serotype. The implications for FMD control of both virus introduction into Uganda and the presence of carrier animals following outbreaks are discussed. PMID- 20003616 TI - Common acute childhood infections and appendicitis: a historical study of statistical association in 27 English public boarding schools, 1930-1934. AB - Although the involvement of common childhood infections in the aetiology of acute appendicitis has long been conjectured, supporting evidence is largely restricted to a disparate set of clinical case reports. A systematic population-based analysis of the implied comorbid associations is lacking in the literature. Drawing on a classic epidemiological dataset, assembled by the School Epidemics Committee of the United Kingdom's Medical Research Council (MRC) in the 1930s, this paper presents a historical analysis of the association between termly outbreaks of each of six common childhood infections (chickenpox, measles, mumps, rubella, scarlet fever and whooping cough) and operated cases of acute appendicitis in 27 English public boarding schools. When controlled for the potential confounding effects of school, year and season, multivariate negative binomial regression revealed a positive association between the level of appendicitis activity and the recorded rate of mumps (beta=0.15, 95% CI 0.07 0.24, P<0.001). Non-significant associations were identified between appendicitis and the other sample infectious diseases. Subject to data caveats, our findings suggest that further studies are required to determine whether the comorbid association between mumps and appendicitis is causal. PMID- 20003617 TI - Bioavailability of procyanidin dimers and trimers and matrix food effects in in vitro and in vivo models. AB - Among procyanidins (PC), monomers, such as catechin and epicatechin, have been widely studied, whereas dimer and trimer oligomers have received much less attention, despite their abundance in our diet. Recent studies have showed that as dimers and trimers could be important in determining the biological effects of procyanidin-rich food, understanding their bioavailability and metabolism is fundamental. The purpose of the present work is to study the stability of PC under digestion conditions, the metabolism and the bioavailability by using a combination of in vitro and in vivo models. Simultaneously, the matrix effect of a carbohydrate-rich food on the digestibility and bioavailability of PC is investigated. The results show a high level of stability of PC under gastric and duodenal digestion conditions. However, the pharmacokinetic study revealed limited absorption. Free forms of dimers and trimers have been detected in rat plasma, reaching the maximum concentration 1 h after oral intake of a grape seed extract. PMID- 20003618 TI - Cross-linguistic syntactic priming in bilingual children. AB - Previous research has used cross-linguistic priming methodology with bilingual adults to explore the nature of their syntactic representations. The present paper extends the use of this methodology to bilingual children to investigate the relation between the syntactic structures of their two languages. Specifically, we examined whether the use of passives by the experimenter in one language primed the subsequent use of passives by the child in the other language. Results showed evidence of syntactic priming from Spanish to English: hearing a Spanish sentence containing a passive led to the increase in children's production of the parallel structure in English. However, there was no priming in the other direction: hearing an English sentence containing a passive did not increase children's use of the parallel structure in Spanish. These results provide evidence for both the integration of syntactic representations in bilingual children and the asymmetry of the relation between their two languages. PMID- 20003619 TI - Differential effects of acute and repeated citalopram in mouse models of anxiety and depression. AB - Clinically, SSRIs are widely prescribed in the treatment of several anxiety disorders, although very few pre-clinical studies have observed a beneficial effect of this class of drugs in animal models of anxiety. Furthermore, the biphasic pattern observed clinically, an exacerbation of anxiety followed by beneficial effects, is rarely observed in animal studies. In the present study we document this clinical phenomenon in several behavioural paradigms. While a single injection of citalopram induced anxiogenic effects, three administrations of citalopram were sufficient to elicit anxiolytic effects. Congruent with these data, we observed that short-term repeated administration of citalopram was accompanied by increased activation of cAMP response element-binding protein (CREB) in the hippocampus and desensitization of 5-HT1A receptors, two phenomena well associated with chronic rather than acute actions of antidepressants. Moreover, effects of citalopram were abolished in CREBalphaDelta mutant animals in the elevated zero maze (EZM) and tail suspension test (TST), but not in novelty-induced hypophagia (NIH). Further, the desensitization of 5-HT1A receptors elicited by citalopram was not affected by CREB deficiency. The significance of the EZM and TST paradigms in predicting therapeutic efficacy is well known while effects in NIH and 5-HT1A sensitization are less well established. These data demonstrate that behavioural responses to citalopram are dependent on the frequency of its administration, and that these responses are differentially dependent on CREB function. PMID- 20003620 TI - Role of the neural cell adhesion molecule (NCAM) in amygdalo-hippocampal interactions and salience determination of contextual fear memory. AB - Evidence suggests that the neural cell adhesion molecule (NCAM) is an important molecular constituent of adaptive and maladaptive circuit (re-)organization in the central nervous system. Here, we further investigate its putative involvement in amygdala and hippocampus functions during context fear memory formation. Using laser capture microdissection and quantitative RT-PCR, we show high NCAM mRNA expression levels in the lateral and basolateral subnuclei of the amygdala, as well as their training intensity- and context-dependent regulation during fear memory consolidation. Moreover, we demonstrate that deficits of NCAM-/- mice in context fear memory can be overcome through contextual pre-exposure, i.e. by reducing the modulatory influence of the amygdala on this hippocampus-dependent memory. On the contrary, NCAM-/- mice failed to increase contextual fear memory after salient overtraining, although they adequately increased their response to auditory-cued fear stimuli. Finally, we demonstrate a reduction of amygdalo hippocampal theta synchronization in NCAM-/- mice during fear memory retrieval. Together, these results suggest an involvement of NCAM-mediated cell recognition processes in information processing of the amygdalo-hippocampal system and in the amygdala-mediated modulation of context fear memory according to stimulus salience. PMID- 20003621 TI - Health effects with consumption of the flax lignan secoisolariciresinol diglucoside. AB - Flaxseed is the richest source of the lignan secoisolariciresinol diglucoside (SDG). After ingestion, SDG is converted to secoisolariciresinol, which is further metabolised to the mammalian lignans enterodiol and enterolactone. A growing body of evidence suggests that SDG metabolites may provide health benefits due to their weak oestrogenic or anti-oestrogenic effects, antioxidant activity, ability to induce phase 2 proteins and/or inhibit the activity of certain enzymes, or by mechanisms yet unidentified. Human and animal studies identify the benefits of SDG consumption. SDG metabolites may protect against CVD and the metabolic syndrome by reducing lipid and glucose concentrations, lowering blood pressure, and decreasing oxidative stress and inflammation. Flax lignans may also reduce cancer risk by preventing pre-cancerous cellular changes and by reducing angiogenesis and metastasis. Thus, dietary SDG has the potential to decrease the incidence of several chronic diseases that result in significant morbidity and mortality in industrialised countries. The available literature, though, makes it difficult to clearly identify SDG health effects because of the wide variability in study methods. However, the current evidence suggests that a dose of at least 500 mg SDG/d for approximately 8 weeks is needed to observe positive effects on cardiovascular risk factors in human patients. Flaxseed and its lignan extracts appear to be safe for most adult populations, though animal studies suggest that pregnant women should limit their exposure. The present review discusses the potential health benefits of SDG in humans, with supporting evidence from animal studies, and offers suggestions for future research. PMID- 20003622 TI - A single dose of vitamin A improves haemoglobin concentration, retinol status and phagocytic function of neutrophils in preschool children. AB - Since there is a reported interrelationship between vitamin A and Fe metabolism, and with immunological response, the objective was to evaluate the effect of a single dose of vitamin A administered to preschool children, on Fe and vitamin A nutritional status, anaemia and phagocytic function of neutrophils, 30 d after supplementation. A total of eighty children (sixty-eight supplemented and twelve controls) were supplemented orally with 200,000 IU (60 mg) vitamin A, and evaluated for nutritional, haematological and immunological responses at the beginning of the study and 30 d after supplementation. Parameters studied included Hb, serum ferritin, retinol and Fe concentrations, transferrin saturation, IL-4, interferon-gamma and phagocityc capacity of neutrophils using non-fluorescent latex microbeads. After supplementation there was a significant increase in Hb concentration (P = 0.03), mean corpuscular Hb concentration (P = 0.001) and serum retinol (P = 0.0078). Prevalences of anaemia and vitamin A deficiency decreased significantly from 17.6 % to 13.2 % and from 25 % to 13.2 %, respectively. Regarding phagocytic function, there was a significant increase in the number of microbeads engulfed by neutrophils (P < 0.05) and no significant changes in cytokine concentrations at 1 month after treatment. A single dose of 200,000 IU (60 mg) vitamin A administered orally to a group of preschool children with a high prevalence of vitamin A deficiency enhanced serum retinol and Hb concentrations, decreased the prevalence of anaemia and vitamin A deficiency and improved the constitutive phagocytic capacity of neutrophils. Vitamin A supplementation could help to decrease vitamin A deficiency, anaemia prevalence and to improve the innate immunity response in preschool children. The effects were obtained without Fe supplementation. PMID- 20003623 TI - Metabolite fingerprinting of urine suggests breed-specific dietary metabolism differences in domestic dogs. AB - Selective breeding of dogs has culminated in a large number of modern breeds distinctive in terms of size, shape and behaviour. Inadvertently, a range of breed-specific genetic disorders have become fixed in some pure-bred populations. Several inherited conditions confer chronic metabolic defects that are influenced strongly by diet, but it is likely that many less obvious breed-specific differences in physiology exist. Using Labrador retrievers and miniature Schnauzers maintained in a simulated domestic setting on a controlled diet, an experimental design was validated in relation to husbandry, sampling and sample processing for metabolomics. Metabolite fingerprints were generated from 'spot' urine samples using flow injection electrospray MS (FIE-MS). With class based on breed, urine chemical fingerprints were modelled using Random Forest (a supervised data classification technique), and metabolite features (m/z) explanatory of breed-specific differences were putatively annotated using the ARMeC database (http://www.armec.org). GC-MS profiling to confirm FIE-MS predictions indicated major breed-specific differences centred on the metabolism of diet-related polyphenols. Metabolism of further diet components, including potentially prebiotic oligosaccharides, animal-derived fats and glycerol, appeared significantly different between the two breeds. Analysis of the urinary metabolome of young male dogs representative of a wider range of breeds from animals maintained under domestic conditions on unknown diets provided preliminary evidence that many breeds may indeed have distinctive metabolic differences, with significant differences particularly apparent in comparisons between large and smaller breeds. PMID- 20003624 TI - Dietary antioxidant restriction affects the inflammatory response in athletes. AB - The purpose of the present study was to determine the effects of dietary antioxidant restriction on plasma concentrations of carotenoids and inflammatory markers at rest and in response to exercise in endurance-trained males. Seventeen males performed two exercise trials 2 weeks apart. Participants followed their habitual antioxidant diet (H-AO) before the first exercise test, then a restricted antioxidant diet (R-AO) for 2 weeks before the second exercise test. Blood was collected pre- and post-exercise. Dietary intakes of fibre, ascorbic acid and beta-carotene were lower (P < 0.05) on the R-AO diet, but no other differences were observed. Pre-exercise plasma beta-carotene concentrations were lower (H-AO, 195 (sd 92); R-AO, 123 (sd 54) ng/ml; P < 0.05), and TNF-alpha concentrations were higher (H-AO, 16 (sd 7); R-AO, 613 (sd 325) pg/ml; P < 0.01) on the R-AO diet compared to the H-AO diet. Most plasma carotenoid concentrations decreased with exercise, but this effect was more consistent on the H-AO diet. No differences in plasma IL-6 concentrations were observed pre-exercise, whereas post-exercise plasma IL-6 concentrations (H-AO, 30.3 (sd 16); R-AO, 15.3 (sd 5) pg/ml; P < 0.05) were lower following the R-AO diet. Post-exercise TNF-alpha concentrations were higher on the R-AO diet. Ratings of perceived effort during submaximal exercise were higher (P < 0.05) on the R-AO diet, but there was no difference in the time to exhaustion between diets. In conclusion, lower dietary intakes of carotenoids alter the plasma concentrations of antioxidants and markers of inflammation at rest and in response to exercise. PMID- 20003625 TI - Diet-induced acidosis: is it real and clinically relevant? AB - The concept of diet-induced 'acidosis' as a cause of disease has been a subject of interest for more than a century. The present article reviews the history of our evolving understanding of physiological pH, the physiological support for the concept of 'acidosis', the causes of acidosis, how it is recognised, its short term effects as well as the long-term clinical relevance of preventative measures, and the research support for normalisation of pH. Further, we suggest differentiation of the terms 'acidosis' and 'acidaemia' as a way to resolve the conflation of these topics which has led to confusion and controversy. The available research makes a compelling case that diet-induced acidosis, not diet induced acidaemia, is a real phenomenon, and has a significant, clinical, long term pathophysiological effect that should be recognised and potentially counterbalanced by dietary means. PMID- 20003626 TI - Placenta: a possible predictor of vitamin A deficiency. AB - The objective of the present study is to assess the association between vitamin A deficiency (VAD) evaluated by serum retinol concentration from the mother and umbilical cord and placental concentration of retinol and carotenoids to propose placental values representative of deficiency. Two hundred and sixty-two puerperal women and their newborns were assessed. Concentration of serum and placental retinol and carotenoids was determined by the spectrophotometric method. Receiver operating characteristic (ROC) curve analysis was performed according to two cut-off points (0.70 and 1.05 mumol/l) to represent deficiency in the placental concentration. No difference between averages of placental retinol and carotenoids was observed in the puerperal women regardless of the cut off point used to define VAD. In relation to the newborns, a decrease (P = 0.012) in placental retinol averages in individuals with VAD was observed when the 1.05 mumol/l cut-off point was adopted. In respect to the placental carotenoid averages, a decrease is observed for both the cut-off points (P = 0.013 and 0.019 for 1.05 and 0.7 mumol/l, respectively). The ROC curve results point to the value of 0.80 mumol/l as representing deficiency with greater values found for sensitivity (66.7 %), specificity (41.7 %) and accuracy (65 %) when the 0.70 mumol/l cut-off point was adopted. The results of the present study show an association between the placental concentration of retinol and carotenoids with clinical VAD, suggesting the need for further studies on more severe cases of deficiency. PMID- 20003627 TI - Is vitamin C supplementation beneficial? Lessons learned from randomised controlled trials. AB - In contrast to the promised 'antioxidant miracle' of the 1980s, several randomised controlled trials have shown no effect of antioxidant supplements on hard endpoints such as morbidity and mortality. The former over-optimistic attitude has clearly called for a more realistic assessment of the benefit:harm ratio of antioxidant supplements. We have examined the literature on vitamin C intervention with the intention of drawing a conclusion on its possible beneficial or deleterious effect on health and the result is discouraging. One of several important issues is that vitamin C uptake is tightly controlled, resulting in a wide-ranging bioavailability depending on the current vitamin C status. Lack of proper selection criteria dominates the currently available literature. Thus, while supplementation with vitamin C is likely to be without effect for the majority of the Western population due to saturation through their normal diet, there could be a large subpopulation with a potential health problem that remains uninvestigated. The present review discusses the relevance of the available literature on vitamin C supplementation and proposes guidelines for future randomised intervention trials. PMID- 20003629 TI - Relationship of frontal lobe dysfunction and aberrant motor behaviors in patients with Alzheimer's disease. AB - BACKGROUND: In order to address the neuropsychological pathogenesis of aberrant motor behaviors in Alzheimer's disease (AD), we used a cross-sectional study design to investigate the association between frontal lobe function, including executive function, and activity disturbances (wandering, purposeless activities and inappropriate activities). METHODS: Among 75 consecutive outpatients with AD, 50 subjects with a Clinical Dementia Rating (CDR) score of 1 or 2 were selected and divided into two groups based on data obtained from interviews with their caregivers: an aberrant motor behaviors (AMB) group (n = 22), and a non-aberrant motor behaviors (NAMB) group (n = 28). Aberrant motor behavior was defined according to whether the "activity disturbance" score (ranging from 0 to 9) of the Behavioral Pathology in Alzheimer Disease (Behave-AD) scale was 0 or >or=1. The total and subtest scores of the Frontal Assessment Battery (FAB) were then compared between the two groups. RESULTS: Significant differences were found between the FAB total (P < 0.05) and the subtest scores (lexical fluency, conflicting instructions; P < 0.05) in the two groups. The FAB score was significantly associated with the activity disturbance score (r = -0.49; P<0.001). A stepwise multiple regression analysis showed that only the FAB score significantly influenced the activity disturbance score (P < 0.001). CONCLUSIONS: This finding suggested that in addition to episodic memory disturbance, frontal lobe dysfunctions might lead patients with AD to develop aberrant motor behavior. PMID- 20003628 TI - Speed of processing training protects self-rated health in older adults: enduring effects observed in the multi-site ACTIVE randomized controlled trial. AB - BACKGROUND: We evaluated the effects of cognitive training on self-rated health at 1, 2, 3, and 5 years post-baseline. METHODS: In the ACTIVE (Advanced Cognitive Training for Independent and Vital Elderly) randomized controlled trial, 2,802 older adults (>or=65 years) were randomly assigned to memory, reasoning, speed of processing, or no-contact control intervention groups. Complete data were available for 1,804 (64%) of the 2,802 participants at five years. A propensity score model was adjusted for attrition bias. The self-rated health question was coded using the Diehr et al. (2001) transformation (E = 95/VG = 90/G = 80/F = 30/P = 15), and analyzed with change-score regression models. RESULTS: The speed of processing (vs. no-contact control) group had statistically significant improvements (or protective effects) on changes in self-rated health at the 2, 3 and 5 year follow-ups. The 5-year improvement was 2.8 points (p = 0.03). No significant differences were observed in the memory or reasoning groups at any time. CONCLUSION: The speed of processing intervention significantly protected self-rated health in ACTIVE, with the average benefit equivalent to half the difference between excellent vs. very good health. PMID- 20003630 TI - Do people become more apathetic as they grow older? A longitudinal study in healthy individuals. AB - BACKGROUND: The aim of this study was to determine levels, rates and progression of apathy in healthy older persons and to investigate factors associated with its progression. METHODS: Seventy-six healthy elderly subjects, aged 58-85 years (mean 69.9), who were recruited by general advertisement and through local community groups, participated as a control group for a longitudinal study of stroke patients. Data were collected on demographic, psychological, neuropsychological and neuroimaging (MRI) variables and apathy was rated by informants on the Apathy Evaluation Scale (AES). RESULTS: Apathy scores and rates increased over 5 years, especially in men. Change of apathy was associated with informant ratings of cognitive decline in the years prior to baseline assessment but not to subsequent neuropsychological, neuroimaging or functional changes. CONCLUSIONS: Apathy increases with age in otherwise healthy community-dwelling individuals, particularly in men. PMID- 20003631 TI - Situational versus chronic loneliness as risk factors for all-cause mortality. AB - BACKGROUND: Several international studies have substantiated the role of loneliness as a risk factor for mortality. Although both theoretical and empirical research has supported the classification of loneliness as either situational or chronic, research to date has not evaluated whether this classification has a differential impact upon mortality. METHODS: To establish the definition of situational vs. chronic loneliness, we used three waves of the Health and Retirement Study (HRS), a nationally representative sample of Americans over the age of 50 years. Baseline data for the present study were collected in the years 1996, 1998, and 2000. The present study concerns the 7,638 individuals who completed all three waves; their loneliness was classified as either not lonely, situational loneliness or chronic loneliness. Mortality data were available through to the year 2004. RESULTS: Those identified as "situationally lonely" (HR = 1.56; 95% CI: 1.52-1.62) as well as those identified as "chronically lonely" (HR = 1.83; 95% CI: 1.71-1.87) had a greater risk for all cause mortality net of the effect of possible demographic and health confounders. Nonetheless, relative to those classified as "situationally lonely," individuals classified as "chronically lonely" had a slightly greater mortality risk. CONCLUSIONS: The current study emphasizes the important role loneliness plays in older adults' health. The study further supports current division into situational vs. chronic loneliness, yet suggests that both types serve as substantial mortality risks. PMID- 20003632 TI - Dental health of community-living older people attending secondary healthcare: a cross-sectional comparison between those with and without diagnosed mental illness. AB - BACKGROUND: Mental illness and cognitive impairment are risk factors for poor dental health. METHODS: We conducted a cross-sectional study to compare the dental health of older patients attending out-patient clinics and day hospitals of old age psychiatry services (the psychiatry group, n = 103) with those attending general/geriatric medical services (the medical group; n = 99). Those living in care homes, and those with diagnosed mental illness (in the medical group) were excluded. A registered mental health nurse assessed mental and general health using validated and previously published instruments. A registered dentist made an independent assessment of dental health (examination to assess oral pathology, status of remaining teeth, and dentures) and made an overall judgment about whether the patient needed any dental treatment (a "normative" need). RESULTS: The normative need for dental treatment was significantly higher among the psychiatry group compared to the medical group (85% vs 52%; p<0.001); even after taking account of the effect of age, gender, teeth status, physical comorbidity, cognition, depressive symptoms, and overall mental and social health [adjusted odds ratio, OR (95% confidence interval): 4.32 (2.09, 8.91)]. The presence of any natural remaining teeth [OR: 4.44 (2.10, 9.42)] and Barthel Index [OR: 0.96 (0.93, 0.99)] were the two other independent predictors of the need for treatment. CONCLUSION: Dental problems are common in community-living older people, especially those with some natural remaining teeth and those with mental illness. There is a need to develop integrated mental health and dental care services for older people with emphasis on prevention of dental problems. PMID- 20003633 TI - Alone? Perceived social support and chronic interpersonal difficulties in suicidal elders. AB - BACKGROUND: Social networks may protect depressed elders against suicidal behavior. However, conflict in important relationships may undermine the sense of social support, potentially negating the protective effects. Thus, we investigated the role of chronic interpersonal difficulties and perceived social support in depressed elders with and without suicidal thoughts and attempts. METHODS: 106 individuals aged 60 years and older participated in this cross sectional, case-control study. They were placed in three groups: suicidal depressed, non-suicidal depressed and non-depressed. Following a detailed clinical characterization, we assessed perceived social support (Interpersonal Support Evaluation List), and chronic interpersonal difficulties (Inventory of Interpersonal Problems). Using general linear models, we explored the relationship between suicidal thoughts/attempts, social support, and chronic interpersonal difficulties. We also examined whether lower perceived social support explained the relationship between chronic interpersonal difficulties and suicidal thoughts/attempts. RESULTS: Suicidal depressed elders reported the lowest levels of perceived social support (belonging, tangible support, and self esteem) and higher levels of chronic interpersonal difficulties (struggle against others and interpersonal hostility), compared to both non-suicidal depressed and non-depressed elders. The relationship between chronic interpersonal difficulties and suicidal behavior was partially explained by low perceived social support. CONCLUSIONS: The experience of strong affects, interpersonal struggle, and hostility in relationships may undermine the sense of social support in depressed elders, possibly leading them to contemplate or attempt suicide. Depressed elders with a history of interpersonal difficulties need to be carefully monitored for suicidal behavior. PMID- 20003634 TI - Developing effective educational approaches for Liaison Old Age Psychiatry teams: a literature review of the learning needs of hospital staff in relation to managing the confused older patient. AB - BACKGROUND: Deficiencies in the knowledge, skills and attitudes of all healthcare professionals working within the general hospital contribute towards the suboptimal care of older hospitalized patients with confusion. In the U.K., policy dictates that Liaison Old Age Psychiatry teams deliver effective education to general hospital clinical staff. The purpose of this paper is to review the literature concerning the learning needs of healthcare professionals in relation to managing confusion in the older patient in order to inform effective educational approaches for Liaison Old Age Psychiatry teams. METHODS: A broad range of medical and educational databases were searched. Identified English language studies were selected for further analysis if they had a specific educational focus in the hospital setting and then further subdivided into intervention and naturalistic studies. The impact of intervention studies was evaluated by Kirkpatrick's system. Learning needs, as determined from the naturalistic studies, were mapped to identify themes. RESULTS: 13 intervention studies were identified. Despite a high level of effectiveness for educational interventions, it was unclear what the active components were. A further 23 naturalistic studies were identified; their findings focused on knowledge gaps, diagnostic behaviors and experiences, attitudes and training issues. Few studies specifically researched learning needs or the educational role of liaison teams. Conspicuous by its absence was reference to relevant educational theories. CONCLUSIONS: The findings of this review can be incorporated in the planning of local curricula by Liaison Teams in order to design educational strategies. There is a need for further research, especially studies exploring the learning needs of all healthcare professionals. PMID- 20003635 TI - Dairy consumption and circulating levels of inflammatory markers among Iranian women. AB - OBJECTIVE: Although recent studies have shown an inverse relationship between dairy consumption and metabolic abnormalities, we are aware of no study evaluating the association between dairy consumption and circulating levels of inflammatory markers. The current study was undertaken to assess the association between the consumption of high-fat and low-fat dairy products and circulating levels of inflammatory markers among Tehrani women aged 40-60 years. DESIGN: In a cross-sectional study of 486 apparently healthy women aged 40-60 years, we assessed usual dietary intakes by means of an FFQ. Anthropometric measurements were made and fasting blood samples were taken for measuring inflammatory markers. RESULTS: The reported mean (SD) daily intake of low- and high-fat dairy consumption was 85 (SD 23) and 101 (SD 29) g/d, respectively. After control for age, BMI, waist circumference and other potential confounders, low-fat dairy consumption was inversely associated with C-reactive protein (beta = -0.04), IL-6 (beta = -0.02) and soluble vascular cell adhesion molecule-1 (beta = -0.06); with further adjustment for dietary intakes, the associations remained significant just for soluble vascular cell adhesion molecule-1 (beta = -0.03). High-fat dairy intake was positively associated with log-transformed values of serum amyloid A (beta = 0.08) and soluble vascular cell adhesion molecule-1 (beta = 0.05), both before and after adjustment for all potential confounding variables. No overall significant associations were found between total dairy consumption and inflammation. CONCLUSIONS: The current study indicates an independent relationship between high-fat as well as low-fat dairy consumption, not total dairy intake, and some markers of inflammation and endothelial dysfunction. Further studies are required to identify responsible components of dairy products and related mechanisms of action. PMID- 20003636 TI - Session 1: Allergic disease: The challenges of managing food hypersensitivity. AB - Food hypersensitivity (FHS) is the umbrella term used for food allergies that involve the immune system and food intolerances that do not involve the immune system. FHS has a huge impact on quality of life and any dietary advice given should aim to minimise this effect. Despite many advances made in diagnosing and managing patients with FHS, the cornerstone of management still remains avoidance of the relevant food. However, a commonly-presenting dilemma in clinical practice is deciding to what extent the food(s) should be avoided. The level of avoidance required is currently based on the type of FHS the patient has, characteristics of the particular food protein and the natural history of the particular FHS. In addition to management of other FHS, management of cow's milk allergy requires the healthcare professional to choose the appropriate formula. Information required by the patient also includes understanding food labels and issues surrounding cross-contamination. In order to ensure that the diet is nutritionally sound, advice should be given about suitable food choices and following a healthy balanced diet, whilst taking into account the dietary restrictions. Practical issues that need to be addressed include going on holiday, travelling and eating away from home. The dietitian plays a crucial role in this process. At present, there are no standardised documents or protocols for the management of FHS and practices differ within and between countries. If adrenaline auto-injectors are prescribed, correct administration should be demonstrated and reviewed on an ongoing basis. PMID- 20003637 TI - Session 3 (Joint with the British Dietetic Association): Management of obesity: Weight-loss interventions in the treatment of obesity. AB - Treatments to induce weight loss for the obese patient centre on the achievement of negative energy balance. This objective can theoretically be attained by interventions designed to achieve a reduction in energy intake and/or an increase in energy expenditure. Such 'lifestyle interventions' usually comprise one or more of the following strategies: dietary modification; behaviour change; increases in physical activity. These interventions are advocated as first treatment steps in algorithms recommended by current clinical obesity guidelines. Medication and surgical treatments are potentially available to those unable to implement 'lifestyle interventions' effectively by achieving losses of between 5 kg and 10 kg. It is accepted that the minimum of 5% weight loss is required to achieve clinically-meaningful benefits. Dietary treatments differ widely. Successful weight loss is most often associated with quantification of energy intake rather than macronutrient composition. Most dietary intervention studies secure a weight loss of between 5 kg and 10 kg after intervention for 6 months, with gradual weight regain at 1 year where weight changes are 3-4 kg below the starting weight. Some dietary interventions when evaluated at 2 and 4 years post intervention report the effects of weight maintenance rather than weight loss. Specific anti-obesity medications are effective adjuncts to weight loss, in most cases doubling the weight loss of those given dietary advice only. Greater physical activity alone increases energy expenditure by insufficient amounts to facilitate clinically-important weight losses, but is useful for weight maintenance. Weight losses of between half and three-quarters of excess body weight are seen at 10 years post intervention with bariatric surgery, making this arguably the most effective weight-loss treatment. PMID- 20003638 TI - Session 4: CVD, diabetes and cancer: Evidence for the use of the Mediterranean diet in patients with CHD. AB - Diet is associated with the development of CHD. The incidence of CHD is lower in southern European countries than in northern European countries and it has been proposed that this difference may be a result of diet. The traditional Mediterranean diet emphasises a high intake of fruits, vegetables, bread, other forms of cereals, potatoes, beans, nuts and seeds. It includes olive oil as a major fat source and dairy products, fish and poultry are consumed in low to moderate amounts. Many observational studies have shown that the Mediterranean diet is associated with reduced risk of CHD, and this result has been confirmed by meta-analysis, while a single randomised controlled trial, the Lyon Diet Heart study, has shown a reduction in CHD risk in subjects following the Mediterranean diet in the secondary prevention setting. However, it is uncertain whether the benefits of the Mediterranean diet are transferable to other non-Mediterranean populations and whether the effects of the Mediterranean diet will still be feasible in light of the changes in pharmacological therapy seen in patients with CHD since the Lyon Diet Heart study was conducted. Further randomised controlled trials are required and if the risk-reducing effect is confirmed then the best methods to effectively deliver this public health message worldwide need to be considered. PMID- 20003639 TI - Plenary Lecture 3: Food and the planet: nutritional dilemmas of greenhouse gas emission reductions through reduced intakes of meat and dairy foods. AB - Legally-binding legislation is now in place to ensure major reductions in greenhouse gas emissions in the UK. Reductions in intakes of meat and dairy products, which account for approximately 40% of food-related emissions, are an inevitable policy option. The present paper assesses, as far as is possible, the risk to nutritional status of such a policy in the context of the part played by these foods in overall health and well-being and their contribution to nutritional status for the major nutrients that they supply. Although meat may contribute to saturated fat intakes and a higher BMI, moderate meat consumption within generally-healthy population groups has no measurable influence on morbidity or mortality. However, high consumption of red and processed meat has been associated with increased risk of colo-rectal cancer and recent advice is to reduce intakes to a maximum of 70 g/d. Such reductions in meat and haem-Fe intake are unlikely to influence Fe status in functional terms. However, overall protein intakes would probably fall, with the potential for intakes to be less than current requirements for the elderly. Whether it is detrimental to health is uncertain and controversial. Zn intakes are also likely to fall, raising questions about child growth that are currently unanswerable. Milk and dairy products, currently specifically recommended for young children and pregnant women, provide 30-40% of dietary Ca, iodine, vitamin B12 and riboflavin. Population groups with low milk intakes generally show low intakes and poor status for each of these nutrients. Taken together it would appear that the reductions in meat and dairy foods, which are necessary to limit environmental damage, do pose serious nutritional challenges for some key nutrients. These challenges can be met, however, by improved public health advice on alternative dietary sources and by increasing food fortification. PMID- 20003640 TI - Mathematical modelling of respiratory syncytial virus (RSV): vaccination strategies and budget applications. AB - We propose an age-structured mathematical model for respiratory syncytial virus in which children aged <1 year are especially considered. Real data on hospitalized children in the Spanish region of Valencia were used in order to determine some seasonal parameters of the model. Weekly predictions of the number of children aged <1 year that will be hospitalized in the following years in Valencia are presented using this model. Results are applied to estimate the regional cost of paediatric hospitalizations and to perform a cost-effectiveness analysis of possible vaccination strategies. PMID- 20003641 TI - Overview of the North American beef cattle industry and the incidence of bovine respiratory disease (BRD). AB - The North American beef cattle industry has endured many changes throughout the years but is encountering some of the most dramatic challenges in history during the first decade of the 21st century. The US beef cattle inventory increased from 1990 to a high of 103.5 million in 1996 and since has declined to a low of 94.5 million on 1 January 2009. Even though economic signals have encouraged the cow herd to increase over the last 5 years, a significant decrease occurred during 2008. It is difficult to determine the precise cost of bovine respiratory disease (BRD) to the industry but it is reported to be greater than US$500 million per year. Data from our practice indicate that losses from BRD over the last 18 years have been characterized by 5-year cycles of decreases and increases. Perhaps it is time for the industry to look for ways to reduce death loss by methods that focus on the animal's response to the pathogens instead of continuing to focus on the pathogens. PMID- 20003642 TI - North American cattle marketing and bovine respiratory disease (BRD). AB - The risk of bovine respiratory disease (BRD) has a significant effect on the value of cattle in the marketplace. Calves sold in larger groups have $6.37/45.45 kg more value than those sold as singles or in small groups. Morbidity is higher in unweaned commingled calves than those marketed in groups more than 45 days following weaning. Calves with an aggressive disposition have significantly less value than docile calves, due largely to depressed performance and less carcass value. The value of cattle in the marketplace can be improved by offering larger, uniform lots of cattle that have been weaned at least 45 days. Cattle that suffer BRD in the feedlot have from $23.23 to $151.18 less value than those remaining healthy. PMID- 20003643 TI - Impact of bovine respiratory disease (BRD) from the perspective of the Canadian beef producer. AB - The costs of bovine respiratory disease (BRD) to the beef producer can be estimated by identifying and summing the direct and indirect costs associated with the disease. The major direct costs are attributable to the cost of the feeder, production costs and carcass disposal. The indirect costs are mainly associated with infrastructure and labour. PMID- 20003644 TI - A dairy producer's view of respiratory disease. AB - Respiratory disease is a significant economic burden for United States dairies. Although there has been quality research, there is a need to translate the academic findings to the dairy barn. In particular, there is a need to implement solutions in the context of the economic ramifications of disease management. Plans to combat respiratory disease need to be simple, implemented incrementally and require low 'management energy'. If respiratory disease is prevented at an early age, this will ensure the greatest chance that the animals will reach their full genetic potential to produce. PMID- 20003645 TI - Challenges and opportunities for managing respiratory disease in dairy calves. AB - Bovine respiratory disease (BRD) is important for the Ontario dairy industry due to the large economic and welfare costs of this disease. Practical science-based management techniques are needed to control and reduce the risk of this disease. Currently, the emphasis on BRD is focused on early detection of disease and prevention. These areas are important but it is not practical to assume this disease will be eliminated in the near future. It is necessary to determine the best practices for caring for sick animals, monitoring their recovery and making changes to their management to facilitate health and recovery. If management changes can be made for animals that are failing to thrive in a current situation, a more complete recovery may be possible and the welfare and economic costs of BRD may be minimized. PMID- 20003646 TI - Genetics, environment and bovine respiratory disease. AB - The heritability estimates for resistance and/or susceptibility to bovine respiratory disease (BRD) are small, suggesting response to direct selection will be slow. The number of mammalian genetic markers associated with resistance to specific pathogens or improved immunity is increasing and will provide additional information for developing selection criteria for producing animals with an innate resistance to BRD. Environmental and management factors play significant roles in the prevalence of BRD and must be considered in a holistic approach to reducing BRD. Although no single solution for preventing BRD is likely to be discovered in the immediate future, the long-term outlook appears very promising. PMID- 20003647 TI - Disease control on organic and natural cattle operations. AB - The number of cattle operations which manage their livestock as certified organic or as natural is constantly increasing. Common conditions that practitioners often see have bacterial, viral or parasite etiologies. With restrictions on treatments allowed, either by the United States Department of Agriculture (USDA) Certified Organic program or by other niche programs, it would help practitioners to know what is allowable to use and how to approach cases in these settings. This presentation aims to provide veterinarians with an awareness of animal health care when working with organic/natural producers and alternative therapies that are understandable to medically trained professionals. PMID- 20003648 TI - Veterinary ethics and production diseases. AB - An animal's welfare should be governed by five freedoms, namely, freedom from hunger and thirst, freedom from discomfort, freedom from pain, injury or disease, freedom to express normal behavior and freedom from fear and distress. If the essence of veterinary medicine is to act like a physician for animals then the profession must be vocal in opposition to production diseases, which can be prevented by changing the system of production. PMID- 20003649 TI - Bovine respiratory disease research (1983-2009). AB - Bovine respiratory disease (BRD) research has provided significant understanding of the disease over the past 26 years. Modern research tools that have been used include monoclonal antibodies, genomics, polymerase chain reaction, immunohistochemistry (IHC), DNA vaccines and viral vectors coding for immunogens. Emerging/reemerging viruses and new antigenic strains of viruses and bacteria have been identified. Methods of detection and the role for cattle persistently infected bovine viral diarrhea virus (BVDV) were identified; viral subunits, cellular components and bacterial products have been characterized. Product advances have included vaccines for bovine respiratory syncytial virus, Mannheimia haemolytica and Pasteurella multocida; the addition of BVDV2 to the existing vaccines and new antibiotics. The role of Mycoplasma spp., particularly Mycoplasma bovis in BRD, has been more extensively studied. Bovine immunology research has provided more specific information on immune responses, T cell subsets and cytokines. The molecular and genetic basis for viral-bacterial synergy in BRD has been described. Attempts have been made to document how prevention of BRD by proper vaccination and management prior to exposure to infectious agents can minimize disease and serve as economic incentives for certified health programs. PMID- 20003650 TI - Host response to bovine respiratory pathogens. AB - Bovine respiratory disease (BRD) involves complex interactions amongst viral and bacterial pathogens that can lead to intense pulmonary inflammation (fibrinous pleuropneumonia). Viral infection greatly increases the susceptibility of cattle to secondary infection of the lung with bacterial pathogens like Mannheimia haemolytica and Histophilus somni. The underlying reason for this viral/bacterial synergism, and the manner in which cattle respond to the virulence strategies of the bacterial pathogens, is incompletely understood. Bovine herpesvirus type 1 (BHV-1) infection of bronchial epithelial cells in vitro enhances the binding of M. haemolytica and triggers release of inflammatory mediators that attract and enhance binding of neutrophils. An exotoxin (leukotoxin) released from M. haemolytica further stimulates release of inflammatory mediators and causes leukocyte death. Cattle infected with H. somni frequently display vasculitis. Exposure of bovine endothelial cells to H. somnii or its lipooligosaccharide (LOS) increases endothelium permeability, and makes the surface of the endothelial cells pro-coagulant. These processes are amplified in the presence of platelets. The above findings demonstrate that bovine respiratory pathogens (BHV 1, M. haemolytica and H. somni) interact with leukocytes and other cells (epithelial and endothelial cells) leading to the inflammation that characterizes BRD. PMID- 20003651 TI - Update on bacterial pathogenesis in BRD. AB - Mannheimia haemolytica, Pasteurella multocida, Histophilus somni, Mycoplasma bovis and Arcanobacterium pyogenes are all frequently implicated in bovine respiratory disease (BRD). M. haemolytica is considered the most important of the group. These bacteria are commensals in the nasopharynx and establish infection in the lungs of cattle that are subjected to a variety of stresses. Factors that permit adherence to and proliferation in the lungs and factors that cause tissue destruction and inflammation have been identified as having major roles in pathogenesis. These virulence factors include protein adhesins, capsular polysaccharide, outer membrane proteins, iron-binding proteins, lipopolysacharide or lipooligosaccharide, enzymes and toxins. These bacterial products function to evade the immune system, damage the immune system and induce a severe inflammatory response. PMID- 20003652 TI - Update on viral pathogenesis in BRD. AB - Many viruses, including bovine herpesvirus-1 (BHV-1), bovine respiratory syncytial virus (BRSV), parainfluenzavirus-3 (PI3), bovine coronavirus, bovine viral diarrhea virus and bovine reovirus, have been etiologically associated with respiratory disease in cattle. This review focuses on the pathogenesis of BHV-1 and BRSV, two very different agents that primarily cause disease in the upper and lower respiratory tract, respectively. PMID- 20003653 TI - Vaccination for respiratory immunity: latest developments. AB - Advances over the last 20 years in immunology and molecular biology have provided many new tools for identifying the important antigens and new ways to achieve the appropriate immune responses to these antigens. These provide many more options to achieve the best immune response from deletion mutations, subunit antigens, vectors or DNA immunization. These tools are being adopted to screen, discover and produce the appropriate antigens and to deliver them by the optimal method and with novel adjuvants to achieve the appropriate immune response. These developments will result in vaccines for respiratory disease that are safer and more efficacious, and provide greater flexibility for use and administration. PMID- 20003654 TI - Antimicrobials and BRD. AB - Pharmacodynamics is limited with respect to its ability to provide precise predictions to guide therapy because of complications related to the bound versus unbound state of the agent, tissue versus plasma concentrations, drug degradation over time, variations among microorganisms, and factors associated with the specific environment at the infection site. Antimicrobial susceptibility testing is likewise imprecise when applied to an individual animal; however, it is valuable on an animal population basis. PMID- 20003655 TI - Future of BRD research: an animal health industry perspective. AB - Industry has made large investments into bovine respiratory disease (BRD) research historically, and will continue to do so, despite the apparent lack of progress, an uncertain regulatory environment, and increased competition for internal resources. Factors such as the growing demand for protein, and the ongoing consolidation and 'technification' of the beef sector globally suggest that the industry will continue to demand interventions that prevent disease, are more efficacious, can be easily administered, and positively affect meat quality. New products must also meet the regulatory requirements of safety and efficacy and anticipate the future needs of the numerous stakeholders in the global food chain. Two obstacles in meeting this challenge are the declining interest in food animal medicine, and BRD specifically, and the reluctance to accept new technology at the consumer level. PMID- 20003656 TI - BRD research needs in the next 10-20 years. AB - Despite numerous advances in bovine infectious disease research, bovine respiratory disease (BRD) remains a significant disease threat and a cause of tremendous loss. Renewed efforts and innovative strategies are required to build on the successes of the past to achieve the successes of tomorrow. Continued and improved support of BRD research will prove to be a worthwhile investment. To achieve better coordination and more efficient use of resources, the following supportive actions should be considered. There should be coordination of integrative teams through an over-arching agency (United States Department of Agriculture (USDA)) to provide peer review and priority review for available public research funds. This would include use of development grants (as seed and matching funding) available to public/private partnerships. Testing and validation of new procedures in production settings (providing more than cost analyses) should be strengthened. Publication of studies should be promoted and old links between clinicians, diagnosticians and basic researchers should be re established. Sets of 'best practices' should be published. Programs to support both basic and applied research through graduate fellowships, residencies, workshops and start-up funding for young research scientists should be strengthened. Sustained funding to train the next generation of research investigators should be addressed as a critical need. PMID- 20003659 TI - Changing of the guard at the CJU. PMID- 20003660 TI - Legends in urology. PMID- 20003661 TI - Endoscopic management of upper tract urothelial carcinoma. AB - While radical nephroureterectomy remains the gold standard of treatment for patients with upper tract urothelial tumors, technological advances have made endoscopic management possible. The careful selection of patients for such an approach is dependent upon an accurate diagnosis and an understanding of the natural history of the disease. High grade tumors behave aggressively and warrant radical extirpation unless an absolute contraindication exists. Motivated patients with low grade tumors and relative contraindications to nephroureterectomy can be managed with percutaneous or retrograde ureteroscopic techniques. High recurrence rates in the ipsilateral upper tract and bladder mandate close surveillance of patients treated conservatively. We review the important diagnostic, staging, technical, and surveillance issues in the endoscopic treatment of upper tract urothelial carcinoma. PMID- 20003662 TI - Is lymphadenectomy indicated in patients with T1 moderately differentiated penile cancer? AB - OBJECTIVE: In patients with penile squamous cell carcinomas (SCCs), lymphadenectomy can be curative and should be considered in cases deemed high risk for metastatic spread to regional lymph nodes. Management of patients without palpable lymphadenopathy remains controversial. Current guidelines for T1 penile SCCs based on previous studies have suggested that moderately differentiated tumors are at low risk for metastatic disease; however given our experience with such patients we sought to examine whether such tumors were truly observable or should be treated more aggressively. MATERIALS AND METHODS: A retrospective chart review of penile cancer cases at three institutions was performed. All slides of patients diagnosed with T1 lesions were rereviewed by our reference pathologists to confirm the original diagnosis and stage. These patients were also reviewed regarding lymphadenectomy results and clinical outcomes. RESULTS: Between 1988 and 2004, a total of 34 cases of SCC of the penis were identified, of which 10 were stage T1. Of these 10 cases, seven had moderately differentiated carcinoma without vascular invasion on pathological evaluation. Metastatic disease was present in one patient at the time of diagnosis and subsequently developed in three of the remaining six patients during follow up. Thus a total of 4 (57%) of the patients developed metastatic disease. CONCLUSIONS: Current management protocols place moderately differentiated T1 penile squamous carcinoma without vascular invasion in a low risk category for metastatic disease. As such, expectant management is currently offered as a primary option for these patients. Our experience suggests that patients in this category are in fact at higher risk for metastatic disease, and may be offered early groin dissection in place of expectant management. PMID- 20003658 TI - Advances and challenges in malaria vaccine development. AB - Malaria remains one of the most devastating infectious diseases that threaten humankind. Human malaria is caused by five different species of Plasmodium parasites, each transmitted by the bite of female Anopheles mosquitoes. Plasmodia are eukaryotic protozoans with more than 5000 genes and a complex life cycle that takes place in the mosquito vector and the human host. The life cycle can be divided into pre-erythrocytic stages, erythrocytic stages and mosquito stages. Malaria vaccine research and development faces formidable obstacles because many vaccine candidates will probably only be effective in a specific species at a specific stage. In addition, Plasmodium actively subverts and escapes immune responses, possibly foiling vaccine-induced immunity. Although early successful vaccinations with irradiated, live-attenuated malaria parasites suggested that a vaccine is possible, until recently, most efforts have focused on subunit vaccine approaches. Blood-stage vaccines remain a primary research focus, but real progress is evident in the development of a partially efficacious recombinant pre erythrocytic subunit vaccine and a live-attenuated sporozoite vaccine. It is unlikely that partially effective vaccines will eliminate malaria; however, they might prove useful in combination with existing control strategies. Elimination of malaria will probably ultimately depend on the development of highly effective vaccines. PMID- 20003663 TI - Editorial comment: The management debate continues! PMID- 20003665 TI - Effect of cranberry drink on bacterial adhesion in vitro and vaginal microbiota in healthy females. AB - INTRODUCTION/OBJECTIVE: Cranberries have been shown to produce urinary metabolites that influence uropathogen adhesion and prevent urinary tract infections. This study was designed to determine if consuming reconstituted, unsweetened cranberry drink from extract retained its bioactive properties by reducing uropathogen adhesion without adversely affecting urinary calcium, magnesium and the vaginal microflora. MATERIALS AND METHODS: A randomized crossover study was undertaken in 12 healthy women consuming reconstituted unsweetened cranberry drink, CranActin or water. The urine was collected at 4 hours and 1 week of consumption and evaluated for antiadhesive properties and urinary pH, calcium and magnesium. Vaginal swabs were collected after 1 week of treatment to assess the vaginal microbiota by DGGE. RESULTS: The resultant urine produced by subjects who consumed 500 ml reconstituted cranberry extract twice per day, significantly reduced the adherence to epithelial cells of P-fimbriated uropathogenic Escherichia coli and showed a tendency towards significance for two E. coli strains expressing fimbriae and an Enterococcus faecalis isolate. The cranberry drink treatment did not alter urinary pH, but reduced calcium and magnesium concentrations compared to water, although not to statistical significance. The reconstituted cranberry drink had no apparent detrimental effect on the vaginal microbiota. However, consuming twice daily resulted in an apparent loss of a potential pathogen from the vagina in 42% subjects. CONCLUSIONS: The present findings suggest that reconstituted cranberry drink may retain the ability to reduce the risk of UTI by inhibiting pathogen adhesion while not detrimentally affecting urinary pH or vaginal microbiota, or the risk of calculi. PMID- 20003666 TI - Fracture risk in androgen deprivation therapy: a Canadian population based analysis. AB - INTRODUCTION: Prostate cancer is the most common noncutaneous malignancy diagnosed in men. The use of androgen deprivation therapy (ADT) is the mainstay of treatment for metastatic disease. The use of ADT has been reported to increase the risk of osteoporosis in men with prostate cancer, with higher risk of fracture than age matched controls. We sought to confirm the higher fracture risk of men with prostate cancer on ADT in the Canadian population. METHODS: We used the Population Health Research Data Repository housed at Manitoba Centre for Health Policy to identify all cases of fractures of the hip, vertebra, or wrist in men aged 50 years and older occurring between 1996 and 2004. Each case was matched with up to three controls by age, sex, ethnicity and medical comorbidity. We calculated the odds ratios (OR) for fracture with prostate cancer, and with or without ADT, after adjusting for possible confounding variables. RESULTS: There were 4696 cases of fracture matched with 14080 controls. After controlling for confounding variables, there was no significant association between prostate cancer and fracture risk (adjusted OR = 0.97, 95% confidence intervals [CI]: 0.83 1.15). We detected a significant association between ADT and fracture risk in men. The adjusted ORs for current and past ADT usage were 1.71 (95% CI: 1.13 - 2.58) and 2.42 (95% CI: 1.42-4.12) respectively. CONCLUSION: Our findings suggest that prostate cancer itself does not increase the risk of fracture and corroborate published results demonstrating an association between ADT and fractures. PMID- 20003667 TI - Multi-session retrograde endoscopic lithotripsy of large renal calculi in obese patients. AB - OBJECTIVES: To establish the safety and efficacy of planned multi-session retrograde endoscopic lithotripsy (REL) for the treatment of large renal calculi in the morbidly obese. METHODS: We retrospectively reviewed charts of patients who underwent multi-session REL procedures from 2003 to 2008. Inclusion criteria included body mass index > 35, total linear stone diameter > 2.0 cm, and patients with a preoperative plan to perform multi-session ureteroscopy. A total of nine patients (six with staghorn calculi) underwent 21 separate procedures. Stone size was measured on preoperative imaging and was defined as length in greatest diameter. Stone free was defined as the complete absence of residual stone on postoperative imaging. RESULTS: Mean body mass index of the patients was 47.8 kg/m2. Mean total linear stone diameter was 3.8 cm. Three of nine patients (33%) were stone free after their final treatment. Mean decrease in stone size from preoperative imaging was 3.3 cm (83%). There were no intraoperative complications. Mean length of follow up was 0.88 years. CONCLUSIONS: Multi session REL is a safe alternative to percutaneous nephrolithotomy (PCNL) in obese patients with very large stones, including staghorn calculi. We recognize that the stone free rate in this series is lower than would be expected with REL for smaller stone burdens or with PCNL. Due to the limitations imposed by both the patient's general medical conditions as well as technical considerations, these patients are left with few options for treatment. Our experience is that management with staged ureteroscopy offers a reduction in stone burden and in some patients a stone free status that provides an acceptable patient outcome. PMID- 20003668 TI - Editorial comment: Re: Multi-session retrograde endoscopic lithotripsy of large renal calculi in obese patients. PMID- 20003669 TI - Renal tumors and the risk of malignancy based on size. AB - PURPOSE: To determine the incidence of malignancy in resected renal tumors in a subpopulation of Canadian patients and the significance of tumor size, patient's demographics, and whether the tumor was an incidental finding. METHODS: Medical records of 168 consecutive nephrectomies performed between March 2003 and June 2008 at our institution were reviewed retrospectively. RESULTS: Average age of the patients was 61 years old (SD 11, range 28-89) and male to female ratio was 1.3:1. Total of 180 masses were resected in 168 nephrectomies (128 radical, 40 partial) during the study period. Of the 180 masses, 20 (11%) were benign and 160 (89%) were malignant lesions. Fifty-five percent of the resected renal masses were incidentally found on preoperative imaging. Based on the pathology reports, the average size of the masses was 5.5 cm (SD 4.0, range 0.3-25.0). The larger masses were more likely to be malignant than the smaller masses (Pearson's chi square test, p = 0.040). CONCLUSION: The present study assists us to adequately assess the risk of malignancy of a renal mass in a Canadian population based on size which allows us to properly advise the patients and suggest best possible treatment options. We recommend more aggressive therapies for masses larger than 4 cm and parenchymal sparing procedures for masses smaller than 4 cm as large proportion of these are benign. PMID- 20003670 TI - Ungated extracorporeal shock wave lithotripsy: safe and effective in the pediatric population. AB - PURPOSE: Ungated extracorporeal shock wave lithotripsy (SWL) is unsynchronized to the patient's electrocardiogram. Although ungated extracorporeal shock wave lithotripsy (SWL) is associated with cardiac arrhythmias in adults, the incidence of arrhythmias in children has not been established. We report on the safety and efficacy of ungated SWL of renal calculi in children. MATERIALS AND METHODS: We evaluated all children less than 18 years of age undergoing ungated SWL for renal calculi. Lithotripsy with gradual incremental energy increase was used to treat the stones. Patients were monitored for arrhythmias and other standard monitoring intraoperatively and postoperatively. RESULTS: Twenty-four consecutive children (10 boys and 14 girls) between 3.5 and 17 years of age underwent 32 ungated SWL procedures for renal calculi. The stone size ranged from 5 mm to 19 mm. No patient had cardiac arrhythmias or other intraoperative complications, required to have the procedure terminated prematurely, or conversion to gated SWL. The overall stone free rate was 87% based on radiographic imaging. CONCLUSIONS: This series supports our initial series that ungated SWL is safe and efficacious in children less than 18 years of age. The arrhythmias associated with adults do no appear to occur in children undergoing ungated SWL. PMID- 20003671 TI - Intravesical therapy use in the high risk patient: practice patterns in an equal access healthcare institution before and after national guidelines. AB - PURPOSE: We examined patterns of intravesical therapy use in nonmuscle invasive bladder cancer over the last 10 years at our institution where there is equal access to healthcare. We further examined any affect that the introduction of national guidelines may have had on the utilization of intravesical therapy in these patients. MATERIALS AND METHODS: An Institutional Review Board (IRB) approved retrospective chart review was performed between the years 1997 and 2007. Only those with premalignant or malignant pathology, as identified using intradepartmental surgical logs and pathology reports, were included. RESULTS: Four hundred seventeen procedures, representing 228 patients, were identified that met the above criteria. A total of 170 high risk, nonmuscle invasive bladder tumors (HG, CIS and T1) were identified, or 41% of cases in whom intravesical therapy was indicated according to the 1999 American Urological Association (AUA) guidelines. One hundred nine (64.2%) received intravesical therapy and 61 (35.8%) did not. This corresponds to an underutilization rate of 19.4% (33/170 high risk tumors did not receive intravesical therapy for unknown reasons). CONCLUSIONS: We have determined that the utilization of intravesical therapy in patients with high risk nonmuscle invasive bladder cancer has improved since the introduction of the 1999 AUA guidelines in an equal access healthcare institution and that patients are compliant with this therapy. PMID- 20003672 TI - Partial ureteropelvic junction obstruction managed by robotic excision and ureteropyelostomy. PMID- 20003673 TI - Laparoscopic management of extensive ureteral fibroepithelial polyps. AB - Fibroepthelial polyps are uniformly benign tumors of the collecting system which may cause obstruction of an affected renal unit. We present a unique case of a 34 year-old male with a solitary functioning kidney who presented with flank pain and renal insufficiency. Radiographic and ureteroscopic evaluation revealed ureteral obstruction due to extensive polyps. After ureteral stenting and normalization of renal function, successful polyp excisions were performed laparoscopically through a ureterotomy. The pathology revealed benign fibroepithelial polyps. The patient remained asymptomatic until 3 years later when ureteroscopy performed for a calculus revealed a widely patent lumen free of polyps. To our knowledge, this is the first published report of a long term follow up after laparoscopic resection of extensive ureteral fibroepithelial polyps. PMID- 20003674 TI - Rectal Hem-o-Lok clip migration after robot-assisted laparoscopic radical prostatectomy. AB - INTRODUCTION: Weck Hem-o-Lok clip migration into the bladder has been reported after robot-assisted laparoscopic radical prostatectomy (RALP). We report a case of Weck clip migration into the rectum presenting as a mass on colonoscopy. METHODS: A 61-year-old male with a prostate specific antigen level of 4.84 ng/ml underwent transrectal ultrasound guided biopsy of the prostate revealing a Gleason's 3 + 3 adenocarcinoma of the prostate involving 20% of the sampled tissue for the left apex. He was subsequently treated with a transperitoneal robot-assisted laparoscopic radical prostatectomy and bilateral pelvic lymphadenectomy. Weck Hem-o-Lok clips were used to ligate the prostate vascular pedicles. The vesicourethral anastomosis was performed using a double armed running technique. RESULTS: Final pathology demonstrated a Gleason 4 + 3 pT2cN0Mx adenocarcinoma of the prostate with negative margins. Four lymph nodes were negative for malignancy. No intraoperative complications occurred. Postoperatively, patient was found to have a Weck Hem-o-Lok clip that migrated into his rectum. This was found on colonoscopy performed for diverticular disease of the colon. The clip was removed without complication. CONCLUSIONS: Judicious use of Weck clips during RALP and communication with physicians participating in patient care for those who have undergone RALP is crucial in minimizing complications and avoiding subsequent procedures. PMID- 20003675 TI - Fistulization in a locally advanced case of squamous cell carcinoma of the prostate. AB - Squamous cell carcinoma of the prostate gland is very rare, constituting 0.5%-1% of all prostatic malignancies. Though it has a similar clinical presentation to prostate cancer, the tumor is more aggressive, spreading to bone, liver and lung. The median survival time is approximately 14 months. Diagnosis is exclusively by histology. Therapeutic options may include radical surgery, radiotherapy, chemotherapy, hormonal therapy or a combination of these treatments. We present a case of locally advanced squamous cell carcinoma of the prostate and comment on its management and subsequent disease related complication. PMID- 20003676 TI - Large bladder diverticulum presenting as an inguinal hernia. AB - We present the case of a 76-year-old man with a large bladder diverticulum presenting as an inguinal hernia with small bowel incarceration. Bladder herniation is extremely rare and when clinically suspected, computed tomography can be an important adjunct to diagnosis. PMID- 20003677 TI - Retroperitoneal ancient schwannoma involving the renal hilum. AB - Schwannomas, a soft-tissue tumor of a Schwann cell, involving the kidney are rare, with few cases available in the literature. Herein, we report a case of a rare variant, an ancient schwannoma of the renal hilum. PMID- 20003678 TI - Small bowel injury during percutaneous nephrostomy tube placement causing small bowel obstruction. AB - Direct small bowel injury is an exceedingly rare complication of percutaneous nephrostomy tube placement. In this report, we present a case of inadvertent injury to the small bowel during percutaneous nephrostomy tube placement with subsequent development of small bowel obstruction. We reviewed the literature to determine the risk factors and appropriate management of small bowel injuries as a result of such a procedure. PMID- 20003679 TI - Testicular infarction as a sequela of inguinal hernia repair. AB - BACKGROUND: Testicular atrophy and necrosis as a result of ischemic orchitis is a well-established complication after inguinal hernia repair. We reviewed four patients with a recent history of inguinal hernia repair and subsequently developed ischemic orchitis to evaluate management options to provide symptomatic relief and prevent testicular atrophy. CASE SERIES: The first patient underwent loosening of inguinal hernia mesh for scrotal pain and decreased arterial testicular flow 8 hours after inguinal hernia repair. The second patient developed ischemic orchitis 20 hours after inguinal herniorrphaphy and required removal of mesh. Neither patient had testicular atrophy at their follow up visit. The third patient underwent orchiectomy for testicular pain and intraoperative findings of infarction. The fourth patient never experienced pain but showed testicular atrophy 6 months after inguinal hernia repair and was managed expectantly. CONCLUSION: In our series, testicular ischemia was reversed in two of the four patients with mesh loosening or removal. Prompt evaluation and exploration of suspected ischemic orchitis after inguinal hernia repair may allow symptomatic relief and prevention of testicular atrophy. PMID- 20003680 TI - Blunt testicular trauma results in rupture of mixed germ cell tumor. AB - It is extremely rare that a documented case of blunt trauma results in rupture of a testicular tumor. We present the case of a 24-year-old man who was crushed by a tree who developed spontaneous testicular rupture. At surgical exploration, he was found to ultimately have a mixed germ cell tumor of the testicle. This case illustrates the importance of physical examination, patient clinical history, and scrotal ultrasound in the management of scrotal trauma. In this instance, the testicular mass ruptured and lead to significant testicular hemorrhage. PMID- 20003681 TI - Squamous cell carcinoma of the renal pelvis with inferior vena cava and iliac vein tumor thrombus. AB - Renal cell carcinoma with inferior vena cava (IVC) tumor thrombus is a well described clinical entity. We report a case of 64-year-old man that developed an aggressive renal pelvic squamous cell carcinoma (SCC) with extensive IVC and bilateral iliac vein tumor thrombus. To our knowledge this is the fifth reported case of renal pelvic SCC with IVC tumor thrombus and the first involving the iliac veins. We review the current therapeutic options for treatment of SCC with IVC involvement. PMID- 20003682 TI - Office based urology trials. PMID- 20003683 TI - Open clinical uro-oncology trials in Canada. PMID- 20003684 TI - Endovascular reperfusion therapy for acute ischemic stroke: a meta-analysis. AB - Intra-arterial thrombolysis has been advocated for treatment of acute ischemic stroke and several prognostic factors were identified in these patients. Other endovascular methods aiming to recanalize occluded vessels including balloon angioplasty, stenting and use of mechanical clot retrieving devices were devised. To date, this approach is replaced by a multi-modal reperfusion therapy (MMRT) approach that emphasizes combination of various methods to achieve reperfusion. MMRT includes intra-arterial infusion of thrombolytics and/or antiplatelet agents, mechanical clot disruption and retrieval and balloon angioplasty with stent placement. Recent results suggest that MMRT results in higher chances for both recanalization of the occluded artery and reperfusion of the ischemic tissue. PMID- 20003685 TI - Recanalization of symptomatic carotid artery dissections causing occlusion with multiple stents: the use of delayed double-contrast road map. AB - BACKGROUND AND PURPOSE: Internal carotid artery dissections (ICADs) with occlusion present with a high morbidity and mortality. No specific medical treatment has proven to be effective in this setting. In selected cases of ICAD with occlusion, stent-assisted angioplasty has been shown to be effective in restoring the perfusion. Spontaneous ICAD causing occlusion successfully recanalized with multiple telescoped stents extending intracranially has only been reported exceptionally. METHODS: We report cases of symptomatic acute carotid occlusion after spontaneous dissection extending from the cervical to the petrocavernous ICA segments. Imaging studies revealed the presence of an extensive penumbra area in every case. Patients were treated by means of multiple stents deployed in a telescoped fashion with the aid of a delayed double-contrast road map. RESULTS: Post-procedural angiography demonstrated restitution of the carotid lumen with no signs of residual dissection or intracranial emboli. The patients improved rapidly, showing no residual neurological deficit after a week. At follow-up, patients are clinically asymptomatic and the vessel is patent with no radiological signs of myointimal hyperplasia. CONCLUSIONS: The successful angiographic and clinical results observed in our cases of extraintracranial stenting of a long carotid dissection causing occlusion contribute to the literature of carotid dissection treated with multiple stents. PMID- 20003686 TI - Is the spinal cord motoneuron exclusively a target in ALS? Comparison between astroglial reactivity in a rat model of familial ALS and in human sporadic ALS cases. AB - OBJECTIVE: Motoneurons are the focus of most investigations of amyotrophic lateral sclerosis (ALS), while the astrocyte reaction is regarded as a phenomenon secondary to neuron degeneration. Since astroglial reactivity differed in different studies of human and animal ALS models and often varied from case to case, we examined and compared astrocyte reactivity within the anterior horns of the spinal cord in a transgenic rat model of familial ALS and in human sporadic ALS (sALS) cases. METHODS: Routine histological staining and immunohistochemical reactions to cytoskeletal proteins [neurofilaments, glial fibrillary acidic protein (GFAP), vimentin and tau] and proliferative markers (proliferating cell nuclear antigen and Ki-67). RESULTS: In human sALS cases and in rats at the early pre-symptomatic and symptomatic stages of the disease, the astroglial reaction was very weak, although there was visible evidence of the morphological manifestations of motoneuron degeneration. Poor immunoreactivity to the GFAP and vimentin antigens and increased expression of tau protein were observed in astrocytes, particularly in the rat model. The astrocyte reaction was evident during a short 'transient' phase of the disease in animals, between the asymptomatic and paretic stages. Proliferating cell nuclear antigen immunoreactivity in glial and neuronal nuclei was observed only in animal material. CONCLUSIONS: Abnormalities in astrocyte cytoskeletal proteins are characteristic features for ALS, both in acquired and congenital forms of the disease. The cytoskeletal aberrations may lead to astroglial dysfunction and disturbances in glutamate uptake that may in turn increase the degeneration of motoneurons. PMID- 20003687 TI - 2010: the year of the lung. PMID- 20003688 TI - Hospital-based surveillance for DR-TB: necessary but not sufficient. PMID- 20003689 TI - Indoor solid fuel combustion and tuberculosis: is there an association? AB - OBJECTIVES: To assess the strength of evidence in published articles for an association between indoor solid fuel combustion and tuberculosis. METHODS: PubMed, a private database and Google Scholar were searched up to May 2008, as was the Cochrane Library (2008, issue 4), to identify articles on the association between indoor air pollution and tuberculous infection, tuberculosis disease and tuberculosis mortality. Each article initially chosen as acceptable for inclusion was reviewed for data extraction by three different reviewers using a standard format. Strength of evidence was determined by pre-determined criteria. RESULTS: The full texts of 994 articles were examined for a final selection of 10 possible articles, of which six met the inclusion criteria. All articles investigated the association between exposure to solid fuel (coal and biomass) smoke and tuberculosis disease. Three (50%) of the six studies included in the systematic review showed a significant effect of exposure to solid fuel combustion and tuberculosis disease-one high-quality case-control study and two cross-sectional studies. CONCLUSION: Despite the plausibility of an association, available original studies looking at this issue do not provide sufficient evidence of an excess risk of tuberculosis due to exposure to indoor coal or biomass combustion. Because the number of studies identified was small, new studies are needed before more definitive conclusions can be reached. PMID- 20003690 TI - Vitamin D receptor genetic polymorphisms and tuberculosis: updated systematic review and meta-analysis. AB - BACKGROUND: Host genetic susceptibility has been suggested as one of the most important explanations for inter-individual differences in tuberculosis (TB) risk. The vitamin D receptor (VDR) gene has been studied as a candidate locus due to genetic polymorphisms that affects the activity of the receptor and subsequent downstream vitamin D-mediated effects. METHODS: We reviewed published studies on VDR polymorphisms and TB susceptibility up to 15 April 2009 and quantitatively summarised associations of the most widely studied polymorphisms (FokI, TaqI, ApaI and BsmI) using meta-analysis. RESULTS: A total of 23 eligible studies were included in this review. Heterogeneous results were observed, which may be partly explained by the differences between populations. Among Asians, the FokI ff genotype showed a pronounced positive association (OR 2.0, 95%CI 1.3-3.2), a significant inverse association was observed for the BsmI bb genotype (OR 0.5, 95%CI 0.4-0.8), and marginal significant associations were found for TaqI and ApaI polymorphisms. However, none of the polymorphisms was significantly related to TB among Africans or South Americans. CONCLUSIONS: The association of VDR polymorphisms with risk of TB observed in our analyses supports the hypothesis that vitamin D deficiency might play a role as risk factor during the development of TB. PMID- 20003691 TI - Drug-resistant tuberculosis in six hospitals in Rio de Janeiro, Brazil. AB - SETTING: Tuberculosis (TB) drug resistance survey in six hospitals in Rio de Janeiro, Brazil. OBJECTIVE: To estimate resistance to at least one drug (DR) and multidrug resistance (MDR) and identify associated factors. DESIGN: One-year cross-sectional survey. Hospitals were included as a convenience sample. RESULTS: Of 595 patients investigated, 156 (26.2%) had previously undergone anti tuberculosis treatment, 433 (72.8%) were not previously treated and information on the remaining 6 was not available. Overall, DR and MDR rates were high, at respectively 102 (17.1%, 95%CI 14.3-20.5) and 44 (7.4%, 95%CI 5.5-9.9) cases. Among individuals not previously treated, 17 had MDR (3.9%, 95%CI 2.4-6.3) and diagnosis in a TB reference hospital was independently associated with MDR (prevalence ratio [PR] 3.3, 95%CI 1.2-8.7) after multivariate analysis. Among previously treated individuals, 27 had MDR (17.3%, 95%CI 11.7-24.2). MDR-TB was independently associated with diagnosis in a TB reference hospital (PR 3.6, 95%CI 1.5-8.7), male sex (PR 2.3, 95%CI 1.2-4.4) and dyspnoea (PR 0.3, 95%CI 0.1-0.7). CONCLUSION: We found high levels of DR- and MDR-TB. Our study design did not permit us to determine the contribution of community versus nosocomial transmission. Further studies are needed to establish this. Nevertheless, hospitals should be recognised as a potential source of transmission of resistant TB strains and urgent measures to avoid nosocomial TB transmission should be taken. PMID- 20003692 TI - Multidrug-resistant pulmonary tuberculosis in Los Altos, Selva and Norte regions, Chiapas, Mexico. AB - OBJECTIVES: To analyse the proportion of multidrug-resistant tuberculosis (MDR TB) in cultures performed during the period 2000-2002 in Los Altos, Selva and Norte regions, Chiapas, Mexico, and to analyse MDR-TB in terms of clinical and sociodemographic indicators. METHODS: Cross-sectional study of patients with pulmonary tuberculosis (PTB) from the above regions. Drug susceptibility testing results from two research projects were analysed, as were those of routine sputum samples sent in by health personnel for processing (n = 114). MDR-TB was analysed in terms of the various variables of interest using bivariate tests of association and logistic regression. RESULTS: The proportion of primary MDR-TB was 4.6% (2 of 43), that of secondary MDR-TB was 29.2% (7/24), while among those whose history of treatment was unknown the proportion was 14.3% (3/21). According to the logistic regression model, the variables most highly associated with MDR TB were as follows: having received anti-tuberculosis treatment previously, cough of >3 years' duration and not being indigenous. CONCLUSIONS: The high proportion of MDR cases found in the regions studied shows that it is necessary to significantly improve the control and surveillance of PTB. PMID- 20003693 TI - An interesting case of rifampicin-dependent/-enhanced multidrug-resistant tuberculosis. AB - We report a case of rifampicin (RMP) dependent/enhanced multidrug-resistant (MDR TB) from a patient who had been treated with the World Health Organization optional thrice-weekly treatment and document the clinical and bacteriological features. RMP-enhanced tubercle bacilli that grew poorly without RMP but grew better in its presence were isolated from the patient with treatment failure. The bacteria grown without RMP consisted of mixed morphologies of short rod-shaped acid-fast bacteria and poorly stained coccoid bacteria, but stained normally as acid-fast rods when grown in the presence of RMP. The isolated RMP-enhanced bacteria harbored the common S531L mutation and a novel mutation F584S in the rpoB gene. Treatment containing RMP or replacement of RMP with more powerful rifapentine paradoxically aggravated the disease, but its removal led to successful cure of the patient. This study highlights the potential dangers of continued treatment of MDR-TB with rifamycins that can occur due to delayed or absent drug susceptibility results and calls for timely detection of RMP dependent/-enhanced bacteria in treatment failure patients by including RMP in culture media and removal of RMP from treatment regimen upon detection. PMID- 20003694 TI - Tuberculosis in asymptomatic HIV-infected adults with abnormal chest radiographs screened for tuberculosis prevention. AB - BACKGROUND: Isoniazid preventive therapy (IPT) prevents tuberculosis (TB) in people living with HIV (human immunodeficiency virus, PLWH). Symptom screening without chest radiographs (CXRs) was established as the strategy for excluding TB disease among PLWH seeking IPT in Botswana's 2001 pilot project. This strategy was evaluated in 2004-2006 among candidates screened for an IPT clinical trial. METHODS: PLWH referred from clinics and HIV testing centers were screened for TB symptoms. All asymptomatic candidates received CXRs; those with abnormal CXRs were investigated further. RESULTS: Among 2732 asymptomatic candidates screened, 302 (11%) had abnormal CXRs potentially compatible with TB; TB disease was diagnosed in 43 of these 302 (14%), or 43 (1.6%) of the 2732 asymptomatic candidates. While not associated with CD4 lymphocyte counts < 200 cells/mm(3), TB was associated with a positive tuberculin skin test (relative risk 2.1, 95%CI 1.1 4.0). IPT was initiated in 113 (62%) of 182 asymptomatic PLWH with abnormal CXRs; 8/113 (7%) subsequently developed TB, and 7/8 (88%) successfully completed anti tuberculosis treatment. CONCLUSIONS: The prevalences of abnormal CXRs and TB were respectively 2.6- and 8.9-fold higher among asymptomatic PLWH screened for the trial than in the pilot. A cost-effectiveness analysis is needed to determine whether the benefits of symptom screening alone are offset by the risk of inducing INH resistance by excluding CXRs during screening. PMID- 20003695 TI - Chest radiograph reading and recording system: evaluation for tuberculosis screening in patients with advanced HIV. AB - SETTING: An antiretroviral treatment (ART) service in Gugulethu township, Cape Town, South Africa. OBJECTIVE: To assess the inter-observer agreement when using the chest radiographic reading and reporting system (CRRS) to detect radiographic abnormalities in patients with advanced human immunodeficiency virus (HIV) associated immunodeficiency being actively screened for tuberculosis (TB). Second, to assess the associated performance characteristics of radiology as a routine screening test for detection of culture-confirmed pulmonary TB. DESIGN: Radiographs from a study in which patients were actively screened for TB just before starting ART were independently reported by two CRRS-certified readers blinded to clinical status. RESULTS: Good kappa statistic agreements between observers were found when reporting any radiological abnormality consistent with TB among all patients (n = 203, kappa = 0.63, 95%CI 0.52-0.73) and among those with culture-confirmed TB (n = 53, kappa = 0.61, 95%CI 0.40-0.83). However, in comparison with sputum culture, the sensitivity (0.68, 95%CI 0.54-0.79) and specificity (0.53, 95%CI 0.45-0.61) of radiology in this patient group were low. CONCLUSION: This study provides evidence of the good inter-observer agreement using the CRRS standardised reporting methodology when used among patients with advanced HIV-associated immunodeficiency and a high prevalence of culture-proven pulmonary TB. The utility of radiology as a screening test for TB in this patient group, however, remains limited. PMID- 20003696 TI - Experience establishing tuberculosis laboratory capacity in a developing country setting. AB - OBJECTIVE: To describe the experience of strengthening laboratory diagnosis of tuberculosis (TB) in a resource-limited country with high TB-HIV (human immunodeficiency virus) and multidrug-resistant TB (MDR-TB) prevalence. METHODS: In the Kingdom of Lesotho, which is confronted with high levels of TB, MDR-TB and HIV prevalence, between 2006 and 2008 a coalition of the Foundation for Innovative New Diagnostics, Partners In Health and the World Health Organization renovated the National TB Reference Laboratory and reinforced microscopy services, streamlined conventional culture and drug susceptibility testing (DST) and introduced modern TB diagnostic methods. FINDINGS: It was feasible to establish a biosafety level three facility for solid culture and DST and an external quality assessment programme for smear microscopy within 4 months, all in 2007. Liquid culture and DST were introduced a month later. Preliminary results were comparable to those found in laboratories in industrialised countries. A year later, line-probe assay for the rapid detection of MDR-TB was introduced. DISCUSSION: Through strong political commitment and collaboration, it is possible to rapidly establish quality assured TB diagnostic capacity, including current methods, in a resource-limited setting. Case detection and management for TB and MDR-TB have been greatly enhanced. From a low baseline, TB culture throughput in the laboratory increased ten-fold and has been sustained. This experience has served as a catalyst to translate policy into practice with new diagnostic technologies. It supports global policy setting to enhance and modernise laboratory work in developing countries. PMID- 20003697 TI - Adding moxifloxacin is associated with a shorter time to culture conversion in pulmonary tuberculosis. AB - OBJECTIVE: To investigate whether adding moxifloxacin (MXF) to the standard anti tuberculosis regimen can shorten the time to sputum culture conversion in pulmonary tuberculosis (PTB). METHODS: Adults with culture-positive PTB were divided into two treatment groups by their choice: standard regimen alone (HERZ group) and standard regimen plus daily 400 mg MXF in the first 2 months (MXF group). Sputum samples were collected thrice weekly in the first 8 weeks. The propensity score was calculated to estimate the conditional probability of entering the MXF group. Factors influencing time to culture conversion were investigated using Cox proportional hazards regression analysis stratified by propensity score. RESULTS: Sixty-two patients were enrolled in the MXF group and 88 in the HERZ group; respectively 51 and 72 completed the study. The regimen was modified before culture conversion in respectively 6 (12%) and 12 (16%; P = 0.47) patients, due to adverse effects. The time to culture conversion was shorter in the MXF group (HR 2.1, 95%CI 1.4-3.2). The culture conversion rate after 6 weeks of treatment was respectively 82% and 61% (P = 0.011, <0.05/4, calculated using the modified Bonferroni method). CONCLUSIONS: Adding MXF to the standard anti tuberculosis regimen in the first 2 months was associated with a shorter time to culture conversion, a higher 6-week culture conversion rate and reduced transmission of tuberculosis. PMID- 20003698 TI - Reduced health provider delay and tuberculosis mortality due to an improved hospital programme. AB - SETTING: A referral hospital in Kaohsiung, Taiwan. OBJECTIVE: To evaluate the impact of an in-hospital tuberculosis (TB) quality care programme initiated in May 2005 on health provider delay and outcome of newly diagnosed TB cases. DESIGN: Retrospective chart review of newly diagnosed TB cases presenting in 2002 and 2006. Health provider delay, clinical manifestations, management and outcome were recorded. RESULTS: Overall, 327 patients before (2002) and 262 patients after (2006) the programme began were enrolled. Patients were older men (mean age 65.9 years) and 23.4% (138/589) had diabetes; 84.4% had received anti tuberculosis treatment. The programme shortened the time for doctors to order a chest X-ray (P < 0.01), and the reporting time for smear (P < 0.0001) and culture (P < 0.0001). On multivariable analysis, risk factors for attributable mortality included age >/=65 years (OR 4.4, 95%CI 1.8-10.9, P = 0.001) and liver cirrhosis (OR 4.3, 95%CI 1.1-16.6, P = 0.04). Treatment reduced mortality by 81% (OR 0.2, 95%CI 0.1-0.4, P < 0.001) and the programme halved overall mortality (OR 0.5, 95%CI 0.3-0.8, P = 0.01), and reduced attributable mortality by 62% (OR 0.4, 95%CI 0.2-0.8, P < 0.01). CONCLUSION: Intervention at the hospital level for quality control of TB care was instrumental in reducing health provider delay and led to a significant reduction in mortality. PMID- 20003699 TI - The impact of nutritional deficit on mortality of in-patients with pulmonary tuberculosis. AB - SETTING: A metropolitan governmental medical centre, Seoul, Republic of Korea. OBJECTIVE: To elucidate the impact of the nutritional deficit assessed by the Nutritional Risk Score (NRS) on the outcomes of in-patients with pulmonary tuberculosis (PTB). DESIGN: All hospitalised patients with microbiologically confirmed PTB were enrolled. A four-point NRS included low body mass index (<18.5 kg/m(2)), hypoalbuminaemia (<30.0 g/l), hypocholesterolaemia (<2.33 mmol/l) and severe lymphocytopaenia (<7 x 10(5) cells/l). The primary outcome was overall in hospital mortality. The degree of radiographical resolution after anti tuberculosis treatment was also evaluated. RESULTS: In a total of 156 patients, the male to female ratio was 1.6:1. The overall mortality was 13.5% and tuberculosis-specific fatality was 3.9%. Predisposing factors and high NRS (> or = 3 points) were independent risk factors for in-hospital death after adjusting for the severity of PTB. High NRS (OR = 16.8, P < 0.001) and predisposing factors (OR = 5.4, P = 0.032) were independent risk factors for 30-day survival. The NRS was not associated with radiographic improvement. CONCLUSION: Regardless of disease severity, the high NRS was a significant negative predictor among in patients with PTB; this finding should therefore be considered in the management of PTB despite the current era of effective anti-tuberculosis chemotherapy. PMID- 20003700 TI - Comparison of the APACHE II, GCS and MRC scores in predicting outcomes in patients with tuberculous meningitis. AB - OBJECTIVE: To evaluate different scoring systems, including Acute Physiology and Chronic Health Evaluation (APACHE) II, the Glasgow Coma Scale (GCS) and the Medical Research Council (MRC) staging system, as well as other prognostic factors, in predicting the discharge outcomes of adult patients with tuberculous meningitis (TBM). DESIGN: We conducted a retrospective analysis of patients admitted with a diagnosis of TBM to a tertiary hospital in northern Taiwan from March 1996 to February 2006. We used APACHE II, GCS, MRC and a variety of factors within 24 h of admission to predict discharge outcomes recorded by the Glasgow Outcome Scale (GOS). RESULTS: Among 43 TBM patients, 33 had a favourable outcome (GOS 4-5), and 10 had an unfavourable outcome (GOS 1-3). The severity of APACHE II, GCS, MRC and presence of hydrocephalus correlated well with the neurological outcomes (P < 0.05). The APACHE II and GCS scoring systems were more sensitive and specific than those of the MRC in receiver operating characteristic analysis. Furthermore, in-hospital mortality could be predicted accurately with APACHE II and GCS. CONCLUSIONS: The APACHE II scoring system is at least as effective as GCS and superior to MRC in predicting the discharge outcomes of adult patients with TBM. PMID- 20003701 TI - The diagnostic efficacy of fine-needle aspiration using cytology and culture in tuberculous lymphadenitis. AB - OBJECTIVE: To retrospectively assess the diagnostic efficacy of fine-needle aspiration (FNA) using cytological and microbiological examinations in tuberculous lymphadenitis. METHODS: Patients with tuberculous lymphadenitis treated at St Mary's Hospital, London, between January 2001 and June 2007 were identified. The cytological and microbiological reports of 97 patients were found. The criteria for a definite diagnosis of tuberculous lymphadenitis were based on a compatible clinical history, tuberculin positivity and either an indicative cytological result or positive culture. RESULTS: In 77 of the 97 (79%) cases, FNA cytology showed evidence of a tuberculous process. In 65 cases, Mycobacterium tuberculosis was cultured from the aspirates, and 54 of these 65 cases showed corresponding cytological evidence of a tuberculous process; 23 cases were diagnosed by cytology but not microbiology, while 11 cases were diagnosed by microbiology but not cytology. CONCLUSION: Cytological and microbiological results appeared to correlate well, but each also gives an exclusive diagnosis. When combining both modalities, the diagnostic efficacy of FNA rises to 91%. A definitive microbiological diagnosis was achieved in 67% of cases and provided information on drug susceptibility. We conclude that samples should be provided for both cytological and microbiological examination when using FNA to diagnose possible tuberculous lymphadenitis. PMID- 20003702 TI - Surgical treatment of non-tuberculous mycobacterial lung disease: strike in time. AB - SETTING: The Netherlands. OBJECTIVE: To describe our experiences with the adjunctive role and benefits of surgery for lung disease due to non-tuberculous mycobacteria (NTM), specifically addressing its indications and timing. DESIGN: Retrospective medical file review of eight patients who underwent surgical treatment for NTM lung disease in the period January 2000 to January 2009, and review of the available literature. RESULTS: Therapy-resistant cavitary NTM disease was the most frequent indication for surgery; two patients underwent pneumonectomy for an infected destroyed lung. Mycobacterium avium was the most common causative agent. Surgery resulted in culture conversion in seven patients; one patient died 2 months after pneumonectomy. No relapses have been noted in the other seven after an average of 19 months of follow-up. CONCLUSIONS: Adjunctive surgical treatment for NTM lung disease yields encouraging results, similar to previously published case series. Careful patient selection, based on extent and type of disease as well as on cardiopulmonary fitness, is important. Potential benefits of surgery should be considered for every individual patient in whom NTM lung disease is diagnosed and re-evaluated after 6 months of treatment. Where possible, surgery should be pursued and conducted in a timely fashion. PMID- 20003703 TI - Demographic risk factors of pulmonary colonization by non-tuberculous mycobacteria. AB - SETTING: British Columbia (BC), Canada. OBJECTIVE: To determine the risk factors for pulmonary colonization by non-tuberculous mycobacteria (NTM). DESIGN: Retrospective study of subjects colonized by NTM from 1990 to 2006. Subjects without mycobacterial disease and with at least three negative cultures served as controls. RESULTS: Mycobacterium avium complex (MAC) species were the most common NTM. Risk factors of colonization included age > or = 60 years (aOR 2.3), female sex (aOR 1.2), residency in Canada for at least 10 years (aOR 3.8), Canadian-born aboriginal (aOR 1.8), and Canadian-born non-aboriginal (aOR 1.4). Predictors of MAC colonization included White race (aOR 1.6) and residency in Canada for at least 10 years, which was the strongest predictor (aOR 6.7). Aboriginal origin was associated with non-MAC colonization (aOR 1.8), and Canadian-born people from the East/South-East Asian ethnic groups were protected from MAC colonization (aOR 0.2), all aOR P < 0.05. CONCLUSION: Older age, female sex, having been born in Canada, long residency in BC and White race predict pulmonary NTM colonization, while Aboriginal origin predicts non-MAC colonization. Further research is needed to identify environmental NTM sources in BC and to determine their relation to colonization and disease. PMID- 20003704 TI - Acceptability and outcome of an Internet-based smoking cessation programme. AB - SETTING: Three district health boards (DHBs), organisations that govern public hospitals and services in Auckland, New Zealand. OBJECTIVE: To evaluate a commercial web-based smoking cessation programme (Smokestop). DESIGN: Smokestop was offered free of charge to 126 staff members of three Auckland DHBs who wanted to stop smoking. Following a 30 minute face-to-face enrolment meeting, participants were able to log on and use the programme. Nicotine replacement therapy (NRT) was available at no cost. All participants who used the programme at least once were followed up at 1, 3 and 6 months after first logging on for assessment of smoking status by self-report verified by carbon monoxide (CO) in expired breath. RESULTS: Of 104 participants who logged onto the programme, 12 (12%) achieved 6-month continuous CO-validated abstinence. Participant feedback was largely positive: 46% agreed that the programme had assisted them and 74% stated they would recommend it to other smokers. The concomitant use of NRT was seen as an important component. CONCLUSIONS: The results suggest that this internet-based smoking cessation programme is an acceptable method to deliver behavioural support to people who want help in stopping smoking, and that it shows promise as a smoking cessation intervention. PMID- 20003705 TI - Drug-resistant tuberculosis epidemic in the Western Cape driven by a virulent Beijing genotype strain. AB - Temporal analysis of drug-resistant tuberculosis (TB) cases in the Western Cape, South Africa, showed a 1.5-fold increase over a 2-year period, suggesting a doubling time of 8.2 years. This increase was strongly associated with multidrug resistance and the Beijing genotype. Forty-two per cent of the overall increase was due to the Beijing genotype strain R220, suggesting that this strain had evolved unique properties that allowed for both acquisition and transmission of drug resistance. To curb the drug-resistant TB epidemic in this setting, it will be essential to implement rapid diagnostics and efficient infection control measures, improve contact screening and ensure treatment adherence. PMID- 20003706 TI - Tuberculosis attributed to household contacts in the Philippines. AB - Data on the burden of disease from tuberculosis (TB) in Filipino households are limited. To determine the magnitude of undiagnosed TB in TB households, and the demographic and socio-economic factors associated with TB in the Philippines, household contacts of adult smear-positive TB patients seen from July 2001 to June 2003 were assessed based on interview, chest X-ray, tuberculin skin test and sputum examination. History of TB and older age were independently associated with TB disease, and age and duration of cohabitation with TB infection. TB and TB infection are highly prevalent in TB households in the Philippines. PMID- 20003707 TI - Hemin toxicity: a preventable source of brain damage following hemorrhagic stroke. AB - Hemorrhagic stroke is a common cause of permanent brain damage, with a significant amount of the damage occurring in the weeks following a stroke. This secondary damage is partly due to the toxic effects of hemin, a breakdown product of hemoglobin. The serum proteins hemopexin and albumin can bind hemin, but these natural defenses are insufficient to cope with the extremely high amounts of hemin (10 mM) that can potentially be liberated from hemoglobin in a hematoma. The present review discusses how hemin gets into brain cells, and examines the multiple routes through which hemin can be toxic. These include the release of redox-active iron, the depletion of cellular stores of NADPH and glutathione, the production of superoxide and hydroxyl radicals, and the peroxidation of membrane lipids. Important gaps are revealed in contemporary knowledge about the metabolism of hemin by brain cells, particularly regarding how hemin interacts with hydrogen peroxide. Strategies currently being developed for the reduction of hemin toxicity after hemorrhagic stroke include chelation therapy, antioxidant therapy and the modulation of heme oxygenase activity. Future strategies may be directed at preventing the uptake of hemin into brain cells to limit the opportunity for toxic interactions. PMID- 20003708 TI - Effect of potent redox-modulating manganese porphyrin, MnTM-2-PyP, on the Na(+)/H(+) exchangers NHE-1 and NHE-3 in the diabetic rat. AB - Increased expression of Na(+)/H(+) exchanger (NHE) and Na(+),K(+)-ATPase activity have been demonstrated in diabetic nephropathy and are implicated in the development of hypertension. The aim of this study was to investigate the effect of a synthetic manganese porphyrin SOD mimic and peroxynitrite scavenger, Mn(III) 5,10,15,20-tetrakis(N-methylpyridinium-2-yl)porphyrin (MnTM-2-PyP) on the expression of NHE and Na(+),K(+)-ATPase activity in the kidneys of streptozotocin (STZ) diabetic rats. MnTM-2-PyP administration (1 mg/kg/day) started immediately after STZ and lasted 2 months. Glucose and glycosylated hemoglobin levels were measured in blood. NHE-1 and NHE-3 isoform expression, Na(+),K(+)-ATPase activity, and markers of ROS/RNS-induced damage were determined in kidney homogenates. Diabetes caused lipid peroxidation, inactivation of aconitase, and increase of nitrotyrosine, which paralleled an increase in NHE-1 and NHE-3 expression and Na(+),K(+)-ATPase activity. MnTM-2-PyP treatment had no effect on blood glucose and glycosylated hemoglobin, but suppressed lipid peroxidation and nitrotyrosine, protected aconitase against inactivation, and reversed the induction of NHE-1 and NHE-3 isoforms. Na(+)/H(+) exchanger is under the control of redox-based cellular transcriptional activity, including members of the SP family of transcription factors. Mn(III) alkylpyridylporphyrins were previously found to inhibit activation of major transcription factors, including SP-1 via scavenging of signaling ROS/RNS. Therefore, our data suggest that, by reducing the levels of ROS/RNS, MnTM-2-PyP might interfere with signaling pathways responsible for NHE up-regulation. PMID- 20003709 TI - Protective effect of selenite on renal ischemia/reperfusion injury through inhibiting ASK1-MKK3-p38 signal pathway. AB - Previous studies have reported that selenite, a known antioxidant, protects brain against ischemia/reperfusion injury, which is mediated by oxidative stress. The aim of this study was to investigate whether selenite can protect kidney against ischemic injury by reducing activation of the apoptosis signal regulating kinase 1 (ASK1)/mitogen-activated protein kinase kinase 3 (MKK3)/p38 mitogen-activated protein kinase signaling pathway. The activation and expression of ASK1, MKK3, p38, caspase 3 and cleaved PARP were analyzed by Western blot. Apoptosis of renal tubular epithelial cells was assessed by the terminal deoxynucleotidyl transferase mediated dUTP nick-end labeling method. Malondialdehyde (MDA) levels were measured by the thiobarbituric acid reaction. Blood serum creatinine and blood urea nitrogen level were measured with an Olympus automatic multi-analyzer. We found that selenite attenuated significantly ASK1, MKK3, and p38 phosphorylation at 3 h after renal ischemia. Furthermore, selenite decreased significantly renal epithelial tubular cell apoptosis. In addition, selenite reduced the MDA level. These findings suggest that the protective action of selenite on ischemia renal injury is associated closely with reducing activation of the ASK1-MKK3-p38 signal pathway. PMID- 20003710 TI - Repeated edaravone treatment reduces oxidative cell damage in rat brain induced by middle cerebral artery occlusion. AB - The free radical scavenger 3-methyl-1-phenyl-2-pyrazolin-5-one (edaravone) has been used to treat acute brain infarction in Japan since 2001. To obtain direct evidence that edaravone serves as an antioxidant in vivo, four groups of rats were prepared: (i) an ischemia/reperfusion (I/R) group receiving 2 h occlusion reperfusion of the middle cerebral artery; (ii) a single administration group treated by intravenous infusion of edaravone (3 mg/kg) immediately after I/R; (iii) a repeated treatment group receiving twice daily edaravone administration for 14 days; and (iv) a sham operation group without occlusion. Repeated treatment with edaravone significantly improved the neurological symptoms and impairment of motor function as compared to the I/R group, while single administration demonstrated limited efficacy. No significant differences in plasma antioxidants such as ascorbate, urate, and vitamin E, or in redox status of coenzyme Q(9) were observed among the four groups. In contrast, the plasma content of oleic acid in the total free fatty acids (percentage 18:1) was significantly increased in the I/R group for 7 days as compared to the sham operation group. Oleic acid was produced from stearic acid by the action of stearoyl-CoA desaturase to compensate for the oxidative loss of polyunsaturated fatty acids. The above results suggest that cellular oxidative damage in the rat brain is evident for at least 7 days after I/R. Repeated treatment suppressed the percentage 18:1 increment, while the single administration did not, which is consistent with the limited efficacy of single administration. PMID- 20003711 TI - Level of nitrated proteins in the plasma, platelets and liver of patients with liver cirrhosis. AB - Over-expression of nitric oxide synthase (NOS) and nitric oxide (NO) formation are associated with the pathogenesis of liver cirrhosis. NO-related stress alters the functions of biomolecules, especially proteins, probably as a result of nitration. The aim of this study was to assess the level of protein nitration and its correlation with the severity of the disease. Liver cirrhosis patients with different grades of severity (grades A, B, and C according to the Child-Pugh classification) were enrolled in this study. Nitroprotein content, arginine, citrulline, NO in terms of total nitrite, nitrosothiol (RSNO) and protein carbonyls were measured in blood. Immunohistochemical detection of nitroprotein was carried out in liver sections of cirrhosis patients. A significant elevation in the levels of serum and platelet arginine, arginase, citrulline, plasma, and platelet nitroproteins, RSNO, total nitrite, protein carbonyls and also a significant amount of nitrated proteins by immunohistochemical detection in tissue were observed in cirrhosis patients. The alterations were highly significant in grade C patients with bleeding complications when compared to those of grade B and A patients. In platelets, both cytosolic and cytoskeletal proteins were found to be nitrated significantly. The level of nitrite seems to have positive correlation with the level of nitroproteins in different grades of cirrhosis. The level of nitroproteins in plasma, platelets and liver tissue can be correlated with the severity of liver cirrhosis. PMID- 20003712 TI - Redox reactivity in propolis: direct detection of free radicals in basic medium and interaction with hemoglobin. AB - Propolis is one of many natural products with known antioxidant properties. The present work aims to investigate the intimate molecular-level mechanisms of this antioxidant reactivity. Electron paramagnetic resonance (EPR)-detectable free radical signals are described here for the first time in propolis extracts. The shape of these signals and the conditions in which they were obtained, point to polyphenolic flavonoids as the sites of the radicals. An inverse correlation between antioxidant capacity and free radical signal intensity is shown. The free radical reactivity of propolis is also illustrated by the effect it exerts on the biologically-relevant peroxide reactivity of hemoglobin. A new test of antioxidant ability in natural extracts such as propolis is proposed, based on modulation of the ascorbate peroxidase activity of hemoglobin (HAPX). Results of this test correlate well with those obtained via traditional methods such as those based on DPPH (2,2-diphenyl-1-picrylhydrazyl), or on ABTS (2,2'-azinobis(3 ethylbenzothiazoline-6-sulfonic acid). PMID- 20003713 TI - Overexpression of Prdx6 and resistance to peroxide-induced death in Hepa1-6 cells: Prdx suppression increases apoptosis. AB - Peroxiredoxins are thiol-specific antioxidants that catalyze the reduction of cellular peroxides and protect cells from ROS-mediated damage and death. Peroxiredoxin gene expression is up-regulated in a number of cancers, suggesting a possible role in cancer cell maintenance. Prdx6, a cytoplasmic protein elevated in certain cancers, is highly expressed in liver and transcriptionally regulated by various oxidative stresses. In the present study, we found that the cancerous Hepa1-6 hepatoma cell line is significantly more resistant to peroxide-induced cytotoxicity than the non-cancerous H2.35 cell line. We also demonstrated that Hepa1-6 cells express approximately 3-fold more Prdx6 mRNA and 2.5-fold more Prdx6 protein than H2.35 cells. Treatment with mithramycin A resulted in a nearly 20% reduction in Prdx6 mRNA in Hepa1-6 cells, suggesting a possible role for Sp1 in Prdx6 up-regulation. We hypothesized that suppression of Prdx6 in Hepa1-6 cells would increase susceptibility to peroxide-induced cell death. Transient transfection of Hepa1-6 cells with Prdx6 siRNA led to a marked reduction in Prdx6 expression, and an increase in peroxide-induced cytotoxicity by apoptosis. Together, these data demonstrate an important anti-apoptotic function for Prdx6 in cancerous liver cells, and suggest that its up-regulation may be a tumor supportive adaptation in cancerous states. PMID- 20003715 TI - Engaging clinicians in quality improvement initiatives: art or science? PMID- 20003714 TI - Influence of cycle exercise on acetone in expired air and skin gas. AB - This study investigated the influence of cycle exercise on acetone concentration in expired air and skin gas. The subjects for this experiment were eight healthy males. Subjects performed a continuous graded exercise test on a cycle ergometer. The workloads were 360 (1.0 kg), 720 (2.0 kg), 990 (2.75 kg) kgm/min, and each stage was 5 min in duration. A pedaling frequency of 60 rpm was maintained. Acetone concentration was analyzed by gas chromatography. The acetone concentration in expired air and skin gas during exercise at 990 kgm/min intensity was significantly increased compared with the basal level. The skin-gas acetone concentration at 990 kgm/min significantly increased compared with the 360 kgm/min (P < 0.05). The acetone excretion of expired air at 720 kgm/min and 990 kgm/min significantly increased compared with the basal level (P < 0.05). Acetone concentration in expired air was 4-fold greater than skin gas at rest and 3-fold greater during exercise (P < 0.01). Skin gas acetone concentration significantly related with expired air (r = 0.752; P < 0.01). This study confirmed that the skin-gas acetone concentration reflected that of expired air. PMID- 20003716 TI - The European Forum for Primary Care and the European Commission consultation process on the European Workforce for Health: some emerging messages. PMID- 20003717 TI - Participants' evaluation of a group-based organisational assessment tool in Danish general practice: the Maturity Matrix. AB - BACKGROUND: The Maturity Matrix is a group-based formative self-evaluation tool aimed at assessing the degree of organisational development in general practice and providing a starting point for local quality improvement. Earlier studies of the Maturity Matrix have shown that participants find the method a useful way of assessing their practice's organisational development. However, little is known about participants' views on the resulting efforts to implement intended changes. AIM: To explore users' perspectives on the Maturity Matrix method, the facilitation process, and drivers and barriers for implementation of intended changes. METHOD: Observation of two facilitated practice meetings, 17 semi structured interviews with participating general practitioners (GPs) or their staff, and mapping of reasons for continuing or quitting the project. SETTING: General practices in Denmark Main outcomes: Successful change was associated with: a clearly identified anchor person within the practice, a shared and regular meeting structure, and an external facilitator who provides support and counselling during the implementation process. Failure to implement change was associated with: a high patient-related workload, staff or GP turnover (that seemed to affect small practices more), no clearly identified anchor person or anchor persons who did not do anything, no continuous support from an external facilitator, and no formal commitment to working with agreed changes. CONCLUSIONS: Future attempts to improve the impact of the Maturity Matrix, and similar tools for quality improvement, could include: (a) attention to matters of variation caused by practice size, (b) systematic counselling on barriers to implementation and support to structure the change processes, (c) a commitment from participants that goes beyond participation in two-yearly assessments, and (d) an anchor person for each identified goal who takes on the responsibility for improvement in practice. PMID- 20003718 TI - Looking inside the black box of community rehabilitation and intermediate care teams in the United Kingdom: an audit of service and staffing. AB - OBJECTIVE: To generate a picture of the range, configuration and staffing of community and intermediate care services in the United Kingdom (UK) and to ascertain whether any relationships exist between service configuration and staffing models. METHOD: A service audit tool was sent to members of the Community Therapist's Network (CTN) and to chief executives of primary care and National Health Service trusts in the UK. Data were collected from the CTN and chief executives of primary care trusts (PCTs) and NHS trusts between late 2005 and early 2006. RESULTS: The overall response rate to the two audits was 37% (n = 243), with 77% of these responses (n = 186) useable. Services varied greatly in terms of their organisation and staffing configurations. Skill mix varied according to the location of service delivery, with home-based services utilising more therapy and support staff than inpatient services. Two clusters of service emerged, based on the number of referrals per year, support staff in the team and the level of care provided by the service. CONCLUSION: There are no clear patterns to the structure and organisation of community and intermediate care services in relation to their purpose, and it remains unclear how different staffing configurations impact on service costs and patient outcomes. The amount of variation observed indicates that there is likely to be considerable variability in service costs and outcomes for the teams. Further evidence is required to determine the impact of different staffing models, and to identify approaches that optimise both effectiveness and efficiency. PMID- 20003719 TI - Are there treatment variations in triage outcomes across out-of-hours co-ops? AB - BACKGROUND: This study considers the factors that affect service provision for individuals who present to out-of-hours (OOH) primary care services in the Republic of Ireland and Northern Ireland. The organisations under consideration are known as OOH co-ops. Specifically, an individual can potentially receive one of four services: nurse advice, doctor advice, a treatment centre consultation or a home visit. AIM: The principal aim was to investigate whether service provision was consistent across co-ops once patient characteristics, patient complaints and other covariates were controlled for. In this paper, service provision was seen as a necessary but not sufficient condition for quality. METHODS: A multinomial logit approach was used to model the choice between the three services offered by co-ops. RESULTS: The results indicate that service provision was relatively homogenous across co-ops. CONCLUSIONS: Quality was consistent across co-ops in terms of service provision. Therefore the next step is to consider whether quality within the treatment received varies. Nevertheless, the result provides some support for using OOH co-ops as a means to provide OOH primary care. PMID- 20003720 TI - Gender makes the difference: the influence of patients' gender on the delivery of preventive services in primary care in Poland. AB - OBJECTIVE: General practitioners' (GPs') time and resources for preventive services needs to be delivered equitably. We aimed to study the effect of patients' gender on the delivery of preventive procedures to adult patients aged 40 years and over. METHOD: An observational study was performed in primary care surgeries in Wielkopolska (Poland) as a part of the Improving Quality in Primary Care (PIUPOZ) programme carried out by Family Medicine Department of the University of Medical Sciences, Poznan. Trained observers directly observed GPs in their office, to register preventive procedures performed during the consultation and in the previous year (via the medical record) in patients aged 40 years and over. RESULTS: A total of 1073 preventive procedures were registered among 450 patients (267 women and 183 men) by 113 doctors in one year. The most common were serum glucose, blood pressure and total cholesterol measurements. Six procedures were offered to less than 10% of patients: dietary advice, tobacco use and alcohol screening, exercise counselling, body mass index (BMI) recording, and waist measurement. Men were more likely to receive tobacco use and alcohol screening and BMI measurement, while more women were offered a total cholesterol screen. CONCLUSIONS: The annual delivery rate of preventive procedures in patients aged 40 years and above is below the recommended level set by the Polish Ministry of Health. Procedures based on clinical examinations or laboratory tests were offered and performed more frequently than lifestyle advice. More men than women received preventive services for tobacco use or alcohol screening and BMI measurements. Patients' gender and physicians' engagement may influence GPs' preventive attitude and performance. These elements should be incorporated in the development of guidelines concerning prevention in primary care. PMID- 20003721 TI - Gender and patient complaints: are they related? AB - BACKGROUND: Several studies suggest that the gender of patients and their healthcare providers affects overall patient satisfaction. AIMS: We sought to determine whether the gender of patients or providers was associated with the number of complaints filed by patients against providers. METHODS: In this case controlled study, complaints from a health maintenance organisation were analysed for gender disparity during a 12-month period. RESULTS: The odds ratio for patients' female gender to be associated with complaints was 3.10 (95% confidence interval 1.73-5.55, P<0.001). Women were also more likely than men to cite providers' behaviour as the cause of complaints. No significant associations were noted between the number of complaints and a complainant's age, diagnosis of chronic pain or psychiatric illness, or with the provider's gender, age or length of employment in the organisation. The heterogeneity of gender effect demonstrated in the literature suggests that this effect is likely to be unique to the different practice settings. CONCLUSION: Evaluating the presence of this phenomenon in the different practice settings is recommended, to improve patient satisfaction and subsequently the quality of care. PMID- 20003722 TI - Antipsychotic prescribing in nursing homes: an audit report. AB - BACKGROUND: In recent years there have been concerns regarding the prescribing of antipsychotic drugs to patients with dementia in nursing homes, due to adverse effects, inappropriate indications and insufficient review. Our practice decided to look at this difficult area more closely. AIM: The aim of this audit was to assess whether our prescribing of antipsychotic drugs in nursing homes was appropriate. METHOD: Our audit criteria for appropriate prescribing were based on National Institute for Health and Clinical Excellence (NICE) recommendations: 1 there is a clear recorded indication or target symptom (standard 100%) 2 there is a documented review of the prescription in the past six months by a GP or psychiatrist (standard 100%). Patients in nursing homes receiving antipsychotic drugs were identified from electronic case notes, using a specific nursing home Read code and British National Formulary chapter. RESULTS: The first data collection (July 2008) revealed we had 81 patients in nursing homes and 22 (27%) were prescribed antipsychotic drugs. There was a clear recorded indication or target symptom in only 11 patients (50%) and a documented review of the prescription in the past six months by a general practitioner (GP) or psychiatrist in only 14 patients (64%). We discussed the results with our practice team and decided to introduce a review checklist for antipsychotic prescribing in nursing homes. The second data collection (December 2008) showed that the number of patients prescribed antipsychotic drugs had fallen to 15 (19% of total 80) and both audit criteria were met in 100% of cases. CONCLUSION: The introduction of the checklist and six-monthly review led to an improvement in the recording of a clear indication for the antipsychotic drug and of documented review. As a practice we were pleased with the modest reduction in the prescribing of antipsychotic drugs in nursing home patients. We were satisfied that the patients who remain on these drugs are being reviewed appropriately and that the benefit of prescribing outweighs the risk. PMID- 20003723 TI - Web alert: involving patients and the public in health service development and research. PMID- 20003724 TI - Spray washing of tomatoes with chlorine dioxide to minimize Salmonella on inoculated fruit surfaces and cross-contamination from revolving brushes. AB - Chlorine dioxide (ClO(2)) is an antimicrobial agent available for commercial produce washing. This study examined the efficacy of ClO(2) at 5 parts per million (ppm) during spray washing of tomatoes (5.0 ml/s per fruit) for preventing Salmonella enterica transfer from inoculated roller brushes to fruit and wash runoff. Furthermore, the sanitizing effects of ClO(2) during spray washing at low and high flow rates (5.0 and 9.3 ml/s per fruit, respectively) on tomato surfaces (nonstem scar areas) with either newly introduced (wet) or overnight air-dried Salmonella inocula were investigated. Salmonella transfer from contaminated brushes to fruit surfaces was reduced 2.1 +/- 0.6 or 4.7 +/- 0.2 log cycles after spray washing with water for 40 s or with the ClO(2) solution for 10 s, respectively. Cross-contamination of Salmonella from brushes to wash runoff during fruit washing for 60 s decreased 5.9 +/- 0.3 log cycles when ClO(2) was used. Fruit washing using contaminated brushes and low flow-rate washing with either water or ClO(2) solution for 10 s reduced newly introduced Salmonella on fruit surfaces by 1.7 +/- 0.6 or 5.1 +/- 0.3 log cycles, respectively. For fruit surfaces with air-dried inocula, washing with water and using uncontaminated brushes for 10 to 40 s reduced Salmonella by 3.2 +/- 0.3 to 3.4 +/- 0.4 log cycles; and the reduction was significantly improved by using ClO(2), high flow rate, or a longer washing time. Washing with ClO(2) at tested flow rates for 10 to 60 s resulted in a 4.4 +/- 0.6 to 5.2 +/- 0.1 log reduction of air-dried Salmonella on fruit surfaces. PMID- 20003725 TI - Use of chemical sanitizers to reduce microbial populations and maintain quality of whole and fresh-cut cantaloupe. AB - Whole cantaloupes either not inoculated or inoculated with Salmonella Poona were submerged in water, 180 ppm of chlorine, acidified calcium sulfate (ACS: 1.2% Safe(2)O-ACS50), 1,000 ppm of acidified sodium chlorite (ASC), 80 ppm of peroxyacetic acid (PAA), and a combination of ACS and PAA for 10 min. Although only ASC and the combination of ACS and PAA significantly reduced the aerobic plate count of samples taken from the surface of whole cantaloupe (compared with samples taken from cantaloupe submerged in water only), all treatments reduced yeast and mold counts on the whole cantaloupe. However, none of the treatments of whole cantaloupes consistently reduced yeast and mold counts for the samples of fresh-cut cantaloupes. The aerobic plate counts for fresh-cut cantaloupe were reduced by 1 to 2 log CFU/g by sanitization of whole fruit with ASC, ACS, and the combination of ACS and PAA. The low bacterial population on the fresh-cut fruit was maintained during 14 days of storage at 4 degrees C. All treatments had a limited effect on the population of Salmonella, achieving no more than a 1.5-log reduction of the pathogen inoculated on the surface of the whole cantaloupes. Salmonella was nondetectable via direct plating (with a detection limit of 0.4 log CFU/g) in fresh-cut cantaloupes prepared from whole cantaloupes treated with any of the sanitizers. However, after enrichment, Salmonella often was detectable. Color, texture, soluble solids, pH, ascorbic acid, and drip loss of cut cantaloupes were not consistently affected by any of the whole-fruit treatments. Overall, treatments of whole cantaloupe with ASC, ACS, and the combination of ACS and PAA at the concentrations tested permitted a significant reduction in Salmonella and native microflora of whole and cut fruit; however, Salmonella still could be found in cut cantaloupes from all treatments. PMID- 20003726 TI - Comparative evaluation of practical functionality of rapid test format kits for detection of Escherichia coli O157:H7 on lettuce and leafy greens. AB - Multistate outbreaks of Escherichia coli O157:H7 infection in 2005 and 2006 associated with fresh and especially minimally processed produce greatly escalated the application of rapid pathogen detection systems to safety management in this food category. Pathogen testing was rapidly integrated into preharvest qualification for field lots, incoming raw produce, or final product. The raw produce and final product were incorporated into test-and-hold programs, typically within a 10-h time frame. To enhance consumer safety and provide guidance for the industry, an assessment of selected kits in comparison to a culture-based method was undertaken. Four primary kits were compared: the Neogen Reveal, SDI RapidChek, BioControl GDS O157, and Qualicon BAX O157 MP. Nine different leafy greens were freshly harvested and inoculated with a five-isolate mixture of E. coli O157:H7 at 10 CFU/25 g of sample, and cultures were enriched following the specified protocol. The PCR method was most consistent for identifying the presence of the inoculated pathogen in the shortest period of time. For the red-pigmented leafy vegetables red butter lettuce, curly endive, red lettuce, and lollo rosa, 13, 38, 88, and 100% false-negative results, respectively, were obtained with the immunoassays, but PCR detection was minimally affected. Immunoassays were negatively affected by delays in achieving critical threshold populations during the allowed enrichment period. Leafy green type, temperature abuse, and preharvest environment were unlikely to affect the results of PCR-based kits. Findings strongly suggest that product testing systems using 8-h detection cutoffs may give false-negative results. These issues become very important in high-throughput testing and retest protocols for presumptive pathogen-positive lots of produce. PMID- 20003727 TI - Heat and drought stress during growth of lettuce (Lactuca sativa L.) does not promote internalization of Escherichia coli O157:H7. AB - Studies were done to determine the effect of heat stress on internalization of Escherichia coli O157:H7 in lettuce subjected to different watering practices during growth. Iceberg and romaine lettuce were grown in sandy soil in an environmental chamber at 23 degrees C during the day and 7 degrees C at night, with a 12-h photoperiod. Thirty days after transplanting seedlings, potting soil was inoculated with a five-strain mixture of green fluorescent protein-labeled E. coli O157:H7 at populations of 4 and 6 log CFU/g of soil. Lettuce plants were exposed to one of two temperature stress regimes: 36 degrees C during the day and 15 degrees C at night for 2 days, or 32 degrees C during the day and 15 degrees C at night for 3 days, both with a 12-h photoperiod. Control plants were held at 23 degrees C during the day and 7 degrees C at night for 3 days. Plants were either watered daily or not watered during the heat stress and control treatments. E. coli O157:H7 was detected by enrichment in all inoculated soil and rhizosphere samples from plants grown in inoculated soil. Less E. coli O157:H7 was detected in inoculated heat-stressed soil than in control soil. From inoculated pots, all leaf surfaces and macerated leaves that had been surface sanitized were negative for E. coli O157:H7. All surface-sanitized macerated roots from control samples and from 143 of 144 samples of inoculated samples were negative for E. coli O157:H7. Heat stress during growth of lettuce did not promote or enhance internalization of E. coli O157:H7, regardless of the moisture content in the soil. PMID- 20003728 TI - Evaluation of two thermal processing schedules at low relative humidity for elimination of Escherichia coli O157:H7 and Salmonella serovars in chopped and formed beef jerky. AB - Foodborne outbreaks have been linked to jerky produced under insufficient thermal processing schedules. Reduction of Escherichia coli O157:H7 and Salmonella serovars during thermal processing of chopped and formed beef jerky was evaluated under two processing schedules representative of those used by large-scale (LS) and small-scale (SS) jerky production facilities. Fresh chopped and formed all beef jerky batter was inoculated with 5.8 to 7.3 log CFU of E. coli O157:H7 or Salmonella per g, extruded into strips, and thermally processed by LS or SS schedules. A >or=5.0-log CFU/g reduction of both pathogens occurred with <10% relative humidity and a cumulative process of 44 min at 55.6 degrees C followed by 46 min at 77.8 degrees C into the LS schedule. Additional drying at 77.8 degrees C for 3.5 h was needed to achieve a water activity of 0.67 and a moisture to-protein ratio (MPR) of 0.77. For the SS process, a >or=5.0-log CFU/g reduction of both pathogens occurred with 15 to 20% relative humidity and a cumulative process of 45 min at 52 degrees C, 60 min at 57 degrees C, 45 min at 60 degrees C, 45 min at 63 degrees C, 90 min at 68 degrees C, and finishing with 30 min at 77 degrees C. After processing for an additional 90 min at 77 degrees C, water activity was 0.60 while the MPR was 0.82. The LS and SS processes for producing chopped and formed jerky provided >or=5.0 log lethality to control E. coli O157:H7 and Salmonella. However, both processes would require additional drying to achieve an MPR of 0.75 to be labeled as jerky. PMID- 20003729 TI - Easy-to-use rapid test for direct detection of Campylobacter spp. in chicken feces. AB - Human campylobacteriosis is the leading cause of acute bacterial gastroenteritis in developed countries. One important source of infection is poultry. Results from the Dutch Campylobacter Risk Management and Assessment project indicate that meat from broiler flocks shedding >or=7 log CFU Campylobacter per g of feces poses the greatest risk of transmitting campylobacteriosis. The objective of this study was to develop a simple and rapid test that would identify chicken flocks shedding high numbers of Campylobacter. We used lateral flow technology as the alternative test method, and selected the culture method according to International Organization for Standardization guidelines. To evaluate the test under field conditions, we sampled either chicken droppings at farms or cecal contents at the slaughterhouse. PCR was used to confirm presumptive Campylobacter spp. colonies. Under laboratory conditions, chicken feces containing >or=6.7 log CFU/g Campylobacter jejuni or >or=7.1 log CFU/g Campylobacter coli were identified by the lateral flow test. Overall, 3 (33%) of 10, and 29 (85%) of 34 C. jejuni- or C. coli-positive chicken flocks were identified at farms and slaughterhouses, respectively, by using the lateral flow test. Fecal samples containing >or=7.3 log of C. jejuni or C. coli CFU/g as determined by plating were always positive when using the lateral flow test. A single person testing seven flocks at a time could obtain test results within 2 h of sampling. This simple and rapid lateral flow test may contribute significantly to the identification of chicken flocks shedding high numbers of Campylobacter. PMID- 20003730 TI - Growth of Cronobacter spp. under dynamic temperature conditions occurring during cooling of reconstituted powdered infant formula. AB - Reconstituted infant formulae are excellent growth media for Cronobacter spp. (formerly Enterobacter sakazakii) and other microorganisms that may be present in such products. Immediate consumption or rapid cooling and storage at a low temperature are therefore recommended as control measures to prevent microbial growth. Placing a container filled with reconstituted liquid formula in the refrigerator, however, does not mean that the temperature of the liquid is directly the same as the set-point of the refrigerator. This study describes the temperature profiles and methods to predict lag time and possible growth of Cronobacter spp. during the cooling process in three types of containers. The overall heat transfer coefficients (alpha) were determined and were shown to have a very large variability in both household refrigerators and an air-ventilated refrigerator equipped with a fan. A mathematical model was built to predict the growth of Cronobacter spp. under dynamic temperature conditions using three models for the lag time. The various estimations for the lag time had a remarkably strong impact on the predicted growth. The assumption of a constant k value (k = lag time x specific growth rate = lambda x micro = 2.88) fitted the experimental data best. Predictions taking into account the large variability in heat transfer showed that proliferation of Cronobacter spp. during cooling may be prevented by limiting the volume to be cooled to portion size only, or by reconstituting at temperatures of 25 degrees C or lower. The model may also be used to predict growth in other situations where dynamic temperature conditions exist. PMID- 20003731 TI - Detection, isolation, and incidence of Listeria spp. in small-scale artisan cheese processing facilities: a methods comparison. AB - Environmental sampling, focused on environmental sites with a high probability of contamination, was conducted in eight artisan cheese processing facilities. Samples (n = 236) from 86 food contact surfaces and 150 non-food contact surfaces were examined for the presence of Listeria spp. by comparing the efficacy of three different primary enrichment media used in five detection and isolation methods. University of Vermont broth was the most sensitive primary enrichment medium for the detection of Listeria spp., including Listeria monocytogenes. These results, however, did not differ significantly from those obtained with Listeria repair broth or Oxoid 24 Listeria enrichment broth. When full methods were considered, the use of dual enrichment methods identified the most Listeria spp.-positive samples, whereas the BAX System PCR Assay for the Genus Listeria 24E provided the most rapid results (approximately 30 h). Cultural results from the direct plating of secondary enrichment broths were generally in agreement with PCR results when compared within methods. Despite minor differences in efficacy, all five methods were in agreement with one another. Overall, 24 (10.7%) of the 236 environmental samples were positive for Listeria spp., all of which were collected from non-food contact surfaces. Nine of these sites were also positive in previous sampling events, suggesting that these sites serve as Listeria niches and that certain ribotypes are particularly persistent, inhabiting environments and specific sites for over 2 years. Overall, our results suggest that the extent of Listeria spp. contamination, notably L. monocytogenes, in small-scale artisan cheese processing environments is low. PMID- 20003732 TI - Anti-Listeria monocytogenes activity of heat-treated lyophilized pomegranate juice in media and in ground top round beef. AB - Heat treatment can affect antimicrobial activity of plant by-products by altering phenolic content and composition and forming melanoidins. The antilisterial efficacy of heat-treated and unheated lyophilized pomegranate juice (LPJ) was determined. The LPJ was heated at 100 degrees C for 0, 30, 60, or 120 min and added at 2% (wt/wt) to ground top round beef, which was then cooked and inoculated with individual L. monocytogenes strains. Samples of meat stored at 5 degrees C were taken at days 1, 8, 14, and 21 and plated onto Oxford medium for enumeration of bacteria. The MIC of LPJ was determined, and agar well diffusion assays were conducted. Against five L. monocytogenes strains, LPJ had a MIC of 1.50 to 1.75% (wt/vol) and 16.8- to 20.0-mm zones of inhibition. In general, no significant differences in L. monocytogenes levels between the various treatments, including the commercial sodium lactate-sodium diacetate combination, were detected at days 1 and 8. The LPJ (0, 30, 60, and 120 min of heating) significantly inhibited growth of all five L. monocytogenes strains in refrigerated ground cooked beef by 1.80 to 4.61 log CFU/g at day 21. Heating did not negatively impact LPJ antilisterial activity. Addition of LPJ lowered pH values by 0.3 units. The L*, a*, and b* values of cooked ground beef with LPJ changed during the study by 3.4 to 4.43, 0.44 to 0.8, and 0.57 to 1.36 units, respectively, compared with the control. This is the first investigation to confirm pomegranate's antilisterial activity in vitro and in ground beef. PMID- 20003733 TI - Inactivation of Listeria monocytogenes on hams shortly after vacuum packaging by spray application of lauric arginate. AB - This study measured and compared the short-term efficacy levels of lauric arginate (LAE) as a postlethality treatment against Listeria monocytogenes present on varied surfaces of large-diameter hams. Preliminary in vitro work demonstrated a 5-log inactivation of L. monocytogenes in 5,000- and 9,090-ppm LAE solutions within 180 min at 4.4 and 23 degrees C. Six different whole-muscle ham types were inoculated with L. monocytogenes at ca. 7-log CFU per ham and spray treated with between 15 and 29 ml of a 9,090-ppm LAE solution, or an equal volume of water (control), prior to vacuum packaging. After 48 h at 4.4 degrees C, populations were recovered from ham and interior packaging surfaces by using a surface rinse method with Dey-Engley neutralizing broth followed by plating on modified Oxford medium. Logarithmic reductions of L. monocytogenes exceeding 2 log CFU/cm(2) of ham surfaces were achieved by LAE treatment on all ham types. Hams with 1,129 cm(2) of surface area that had been processed by drenching in liquid smoke had 3.84 and 2.67 CFU/cm(2) 48 h following treatment with 18 ml of water or LAE, respectively, but increasing treatment volumes to 22 ml significantly reduced (P < 0.05) L. monocytogenes levels to 0.65 log CFU/cm(2). This study demonstrated the efficacy of LAE against L. monocytogenes on several ham types, thereby validating it as a postlethality treatment for inactivation of the pathogen. PMID- 20003734 TI - Purification and partial amino acid sequence of pentocin 31-1, an anti-Listeria bacteriocin produced by Lactobacillus pentosus 31-1. AB - Pentocin 31-1, an anti-Listeria bacteriocin produced by Lactobacillus pentosus 31 1 from the traditional Chinese fermented Xuan-Wei ham, was successfully purified by the pH-mediated cell adsorption-desorption method and then purified by gel chromatography with Sephadex G-10. The purification resulted in a 1,381.9-fold increase in specific activity with a yield of 76.8% of the original activity. Using Tricine-sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS PAGE), the molecular mass of the purified peptide was found to be between 3,500 and 6,400 Da, and bacteriocin activity was confirmed by overlayer techniques. When subjected to mass spectrometry analysis, the protein was highly pure and its molecular mass was 5,592.225 Da. The partial N-terminal sequence of pentocin 31-1 was the following: NH2-VIADYGNGVRXATLL. Compared with the sequence of other bacteriocins, pentocin 31-1 has the consensus sequence YGNGV in its N-terminal region, and therefore it belongs to the class IIa of bacteriocins. PMID- 20003735 TI - Effect of storage and subsequent reheating on viability of Listeria monocytogenes on pork scrapple. AB - We evaluated the fate of Listeria monocytogenes on commercial pork scrapple, a regionally popular, ready-to-eat (RTE) meat. We also conducted an informal survey to address consumer practices for storing and reheating scrapple. Of the 129 consumers who responded to at least one of the eight questions posed in the survey, about half (46.4%; 52 of 112) considered scrapple RTE, the majority (69.7%; 76 of 109) stored it in the refrigerator, and all (100%; 112 of 112) preferred to reheat it prior to consumption. Most respondents (83.9%; 94 of 112) reheated the scrapple by pan frying for 1 to 10 min at medium to high temperature. To study pathogen behavior, slices of pork scrapple were surface inoculated with a five-strain cocktail of L. monocytogenes (ca. 2.0 log CFU/g), vacuum sealed, and stored for up to 60 days. Pathogen levels increased to 8.9, 9.5, and 9.9 log CFU/g after 44 (4 degrees C), 21 (10 degrees C), and 5 (21 degrees C) days, respectively. When slices 1.3 cm (ca. 55 g) and 1.9 cm (ca. 85 g) thick were surface inoculated with L. monocytogenes (ca. 7.0 log CFU/g) and then reheated in a skillet (191 degrees C) for 0.5 to 4 min per side or to target instantaneous internal temperatures of 48.9 to 71.1 degrees C, it was possible to achieve pathogen reductions ranging from ca. 2.2 to 6.5 log CFU/g. These data confirm that in the unlikely event of postprocessing contamination of pork scrapple by L. monocytogenes, proper reheating can appreciably reduce levels of the pathogen before consumption. PMID- 20003736 TI - An immunomagnetic PCR signal amplification assay for sensitive detection of Staphylococcus aureus enterotoxins in foods. AB - Enterotoxigenic strains of Staphylococcus aureus produce a variety of heat-stable staphylococcal enterotoxins (SEs) that are a prevalent cause of food poisoning in the United States and other countries. Many immunological and biochemical assays often work well in buffer systems but are hindered when tested in the complex chemical environment of foods. To overcome these biases and improve the limits of detection, we implemented an immunomagnetic PCR signal amplification assay (iPCR SA) for recovery and detection of SEA and SEB in foods. Anti-SEA or anti-SEB primary antibodies were coated onto COOH-modified magnetic beads using 1-ethyl-3 (3-dimethyl aminopropyl) carbodiimide reagent. Secondary antibodies were covalently linked to amino-modified reporter DNA oligonucleotides (563 bp) via the linker molecule succinimidyl-4[N-maleimidomethyl]-cyclohexane-1-carboxylate. An internal 159-bp portion of the reporter DNA retained by the captured toxin molecule was then amplified by real-time PCR. A semiautomated Bead Retriever proved extremely helpful in both the application of the conjugation chemistries and required washes and the recovery and washing of bead-conjugated toxin from tested food samples. The procedure was simple, and analyses were completed in 5 to 6 h. The assay was sufficiently robust that we were able to detect SEA and SEB in tryptic soy broth, milk, lemon cream pie, tuna salad, deli turkey, and ground turkey at levels as low as 7.5 fg/ml. SE was still detected at high sensitivity after heating in food samples for typical pasteurization or cooking regimens. Sensitivity was diminished only when samples were subjected to extreme heating. PMID- 20003737 TI - Hollow-fiber ultrafiltration for the concentration and simultaneous recovery of multiple pathogens in contaminated foods. AB - We investigated the possibility of using hollow-fiber ultrafiltration (HUF) for the simultaneous recovery of multiple microorganisms in food samples. MS2 bacteriophage, E. coli, Bacillus subtilis spores, and murine norovirus (MNV) were each inoculated into 5 liters of either distilled water (DW) or glycine elution buffer and then concentrated using hollow-fiber polysulfone ultrafilters. The resulting concentrates were further analyzed by either cultivation or TaqMan real time reverse transcription PCR assay. The overall average recovery rates were 7.1% in DW and 17.1% in glycine elution buffer. When the virus, vegetative bacteria, and bacterial spores were simultaneously inoculated into DW, glycine, or Tris-HCl elution buffers, on average 16.8% of inoculated microorganisms were recovered by HUF. The addition of 3% beef extract blocking buffer to HUF increased the total recovery rate to 46.1%, with incremental recovery rates increasing sharply for B. subtilis spores and MNV. Use of HUF resulted in E. coli recovery rates of 68.0% on lettuce and 66.2% on ham and MNV recovery rates of 1.5% on lettuce and 5.8% on ham. Our study demonstrates that HUF can be effective at simultaneously recovering and concentrating diverse bacterial and viral pathogens from foods. PMID- 20003738 TI - An integrated approach to extend the shelf life of a composite pastry product (cannoli). AB - In this study, a combined approach is proposed to extend the shelf life of a composite pastry product (cannoli). In particular, to delay moisture migration, one, two, or three layers of a zein-based coating were studied. A three-layer coating represented the most effective solution to prevent rapid pastry softening. A subsequent experimental trial was aimed to prolong the shelf life of the ricotta-based stuffing. To this aim, two different antimicrobial compounds (lysozyme and lemon extract) at three concentrations (2,000, 3,000, and 4,000 ppm) were investigated separately from a microbiological and a sensorial point of view. Lemon extract was the active compound that received a better score, thus suggesting using 2,000 ppm of citrus extract in the last step. In the final experimental trial, cannoli were coated with three layers of zein, stuffed with ricotta containing the selected active agent, and packaged in two microperforated films. The use of zein-based coating and the lemon extract in the ricotta stuffing, combined with the barrier properties of the selected packaging materials, allowed a significant prolongation of cannoli shelf life, regardless of the type of film: a shelf life of more than 3 days was recorded, compared with the control samples, which were acceptable for less than 2 days. It is reasonable to assume that the proposed integrated approach could boost the distribution of the investigated typical pastry beyond local borders. PMID- 20003739 TI - Inactivation of fungal spores in apple juice by high pressure homogenization. AB - High pressure homogenization (HPH) at 300 MPa in apple juice provides more than 5 log kill of ascospores of Saccharomyces cerevisiae, conidiospores of filamentous fungi, and sporulated black yeasts. HPH and heat treatment were more effective against vegetative cells than against the spores of yeasts used in this study. Ascospores of Talaromyces macrosporus and Neosartorya spinosa were resistant to HPH at 300 MPa. HPH of ascospores of T. macrosporus may result in activation. PMID- 20003740 TI - Survival of North American genotypes of Trichinella in frozen pork. AB - North American genotypes of Trichinella spiralis (T-1), Trichinella nativa (T-2), Trichinella pseudospiralis (T-4), Trichinella murrelli (T-5), and Trichinella T-6 were examined for susceptibility to freezing in pork using time-temperature combinations that have been proven to inactivate T. spiralis. Infections were established in 3-month-old pigs of mixed sex and breed by oral inoculation of 10,000 muscle larvae (ML) (all genotypes, rodent-derived ML), 20,000 ML (T-1, T 4, and T-5; cat-derived ML), or 30,000 ML (T-2 and T-6; cat-derived ML). Pigs were euthanized 60 days postinoculation. Muscles from the tongue, masseter muscles, diaphragm, triceps, hams, neck, rump, and loins were ground, pooled, and mixed to ensure even distribution of larvae. Samples (20 g) containing each Trichinella species, genotype, and source combination were placed in heat sealable pouches, transferred to a constant temperature refrigerant bath, and maintained according to defined time and temperature combinations. Larvae recovered from cold-treated pork samples were inoculated into mice to determine infectivity. Results indicated that the time-temperature combinations known to render pork safe for T. spiralis are sufficient to inactivate T. nativa and T-6 (the freeze-resistant isolates), T. murrelli (the most common sylvatic species in the United States excluding Alaska), and T. pseudospiralis (a species that lacks a muscle nurse cell). These data close a gap in knowledge about the effectiveness of freezing for inactivating these parasites in pork and should alleviate concern about the safety of frozen pork products from the United States. PMID- 20003741 TI - Migration of bisphenol A from can coatings to liquid infant formula during storage at room temperature. AB - Information on migration of bisphenol A (BPA) from can coatings to foods during storage at room temperature is very limited, and the conclusions from the available studies are not always consistent. To investigate the effect of storage time on BPA migration from can coatings to liquid infant formula at room temperature, samples of 21 canned liquid infant formula products from different cans but the same lot as those analyzed for BPA previously were analyzed for BPA again after storage at room temperature for 10 months. Additional migration of BPA from can coatings to liquid formula during the 10-month storage period at room temperature was observed for 9 of the 21 products, with increases in BPA levels ranging from 29.8 to 110%. Significant differences between the 2007 and 2008 results (P = 0.026) were observed for only one brand of product, which had the lowest BPA levels in the 2007 survey. The BPA levels in the milk-based formula products analyzed in 2008 (mean, 6.8 ng/g) were significantly higher (P = 0.00023) than those in the milk-based formula products analyzed in 2007 (mean, 5.0 ng/g), whereas the differences in BPA levels between the soya-based formula products analyzed in 2008 (mean, 5.3 ng/g) and those analyzed in 2007 (mean, 5.8 ng/g) were not significant (P = 0.097). No obvious correlation between the product expiration date and the level of BPA migration from can coatings was observed. PMID- 20003742 TI - High-risk food consumption and food safety practices in a Canadian community. AB - Understanding consumers' high-risk food consumption patterns and food handling in the home is critical in reducing foodborne illness. This study was conducted to determine the prevalence of unsafe food practices of individuals in a Canadian based population, specifically, high-risk food consumption and home food safety practices. During November 2005 to March 2006, a sample of 2,332 randomly selected residents of the Waterloo Region (Ontario, Canada) participated in a telephone survey of food consumption and food safety. Questions covered consumption of high-risk foods, hand washing practices, safe food handling knowledge, source of food safety education, meat thawing and cooking practices, cross-contamination after raw food preparation, and refrigeration temperatures. Certain high-risk food behaviors were common among respondents and were associated with demographic characteristics. In general, unsafe practices increased with increasing total annual household income level. Males were more likely to report engaging in risky practices than were females. Specific high risk behaviors of public health concern were reported by elderly individuals (e.g., consuming undercooked eggs), children (e.g., consuming chicken nuggets), and rural residents (e.g., drinking unpasteurized milk). Respondents appeared to know proper food safety practices, but did not put them into practice. Thus, educational programs emphasizing specific practices to improve food safety should be directed to targeted audiences, and they should stress the importance of consumer behavior in the safety of foods prepared at home. Further investigation of consumer perceptions is needed to design such programs to effectively increase the implementation of safe food practices by consumers. PMID- 20003743 TI - Influence of beta-agonists (ractopamine HCl and zilpaterol HCl) on fecal shedding of Escherichia coli O157:H7 in feedlot cattle. AB - Ractopamine HCl and zilpaterol HCl, beta-agonists recently approved for use in feedlot cattle to improve performance traits and carcass leanness, were examined for their effects on fecal shedding of Escherichia coli O157:H7 in feedlot cattle. Fecal samples (n = 2,454) were obtained from four experiments (one ractopamine HCl, three zilpaterol HCl) over the course of a 3-year period, either by rectal palpation (ractopamine HCl experiment) or from pen-floor fecal pats. Samples were cultured quantitatively and qualitatively for E. coli O157:H7. No significant treatment differences were detected for fecal prevalence of E. coli O157:H7 in the ractopamine HCl experiment. Zilpaterol HCl feeding had no effect (P > 0.20) on fecal shedding in the first or second experiments, with overall E. coli O157:H7 prevalence relatively low (<7%). In the third zilpaterol HCl experiment, the percentage of fecal samples that were E. coli O157:H7 positive following qualitative culture was higher (P < 0.05) in the zilpaterol HCl treatment (10.3%) than for the control (6.1%). The current research showed minimal effects of beta-agonists on fecal shedding of E. coli O157:H7 and indicated that these compounds (fed immediately prior to slaughter) are not a cause for concern from a food safety standpoint. PMID- 20003744 TI - Use of cellulose filters to isolate Campylobacter spp. from naturally contaminated retail broiler meat. AB - Membrane filtration has been used to isolate Campylobacter spp. from feces, although approximately 5 log CFU/g must be present in the sample. Few studies have attempted to use filter membranes for the isolation of Campylobacter from foods. We investigated the minimum number of thermotolerant Campylobacter cells that pass through cellulose filters, the effect of different cell conditions on the rate of passage, and the minimum number of cells that could pass the filters from enriched broiler meat naturally contaminated with Campylobacter spp. Cellulose filters with 0.65-microm pore sizes retained fewer cells and were more effective than filters with 0.45-microm pore sizes. Scanning electron microscopy revealed that 15 min of contact of the filters with agar plates allowed for the passage of most bacteria. The minimum number of bacteria required to pass through the filters was contingent on cell conditions; nonmotile cells were retained more than motile cells (P < 0.05). The minimum number of motile bacteria from 24-h cultures and centrifuged cells were 2.2 and 2.1 log CFU, respectively, while the number of coccoid and nonmotile (flaA/B(-) mutant) cells were 4.1 and 3.4 log CFU, respectively. Broiler meat samples enriched in Bolton's broth supplemented with 5% lysed blood showed that approximately 1.7 log CFU of Campylobacter can be filtered to pure colonies on agar plates. These results demonstrate that the motility of the bacteria influences passage through cellulose filters and that 0.65-mum-pore-size filters on agar plates help obtain pure Campylobacter colonies from enriched food samples. PMID- 20003745 TI - Pseudomonas fragi strains isolated from meat do not produce N-acyl homoserine lactones as signal molecules. AB - Quorum sensing (QS) is a signalling mechanism through which bacteria cellular functions are modified to promote access to nutrients and more favorable environmental niches. The frequent occurrence of Pseudomonas spp. in fresh and spoiled meat may involve enhanced gene expression regulated by QS. Several Pseudomonas spp. produce different N-acyl homoserine lactone (AHL) signal molecules. Meat spoilage during aerobic, refrigerated storage is often associated with the presence of Pseudomonas fragi. As with other Pseudomonas species in natural habitats, the dominance and activities of P. fragi in meat may be regulated by QS. In this study, five biosensor strains were used to detect AHL production on three different media by 72 different P. fragi strains isolated from fresh and spoiled meat. Positive and negative AHL-producing strains were used to verify the assays. None of the strains produced detectable quantities of AHLs, even when concentrated cell-free culture supernatants were assayed, nor did exogenous lactones increase biofilm formation in P. fragi strains. However, all isolates produced furanosyl borate diesters (type II autoinducers; AI-2) when tested using the bioluminescent biosensor strain of Vibrio harveyi (BB170). The production of AI-2 was presumed to be of metabolic origin even though Pseudomonas spp. have not been shown to harbor the luxS gene. Thus, the efficient development of P. fragi in fresh meat is not regulated by an AHL-mediated QS system. The mechanism of AI-2 production and its possible role in spoilage dynamics needs further study. PMID- 20003746 TI - Chicken salad as the source of a case of Listeria monocytogenes infection in Connecticut. AB - Listeriosis is a severe infection with high morbidity and mortality. We report a fatal case of listeriosis in a patient with a history of Crohn's disease who consumed chicken salad purchased from a retail food establishment before developing listeriosis. As part of the regulatory testing programs, the U.S. Department of Agriculture Food Safety and Inspection Service and the Florida Department of Agriculture and Consumer Affairs found that chicken products from a single food-production establishment were contaminated with Listeria monocytogenes, resulting in a product recall. The case patient's Listeria isolate was subtyped by pulsed-field gel electrophoresis (PFGE) and matched the Listeria isolates from the recalled chicken products. Identification of the source of Listeria involved collaboration among two state public health laboratories and epidemiologists and state and federal regulatory agencies. PFGE typing can be used to reveal correlations between clusters of human illness and contaminated food products and to rapidly identify sources of Listeria infection to allow implementation of corrective actions at both the state and national levels. PMID- 20003747 TI - Microbial quality of soft drinks served by the dispensing machines in fast food restaurants and convenience stores in Griffin, Georgia, and surrounding areas. AB - This study was undertaken to evaluate the microbial quality of the soft drinks served by fast food restaurants and gas station convenience stores in Griffin, GA, and surrounding areas. The soft drinks were collected from the dispensing machines in 8 fast food restaurants or gas station convenience stores in 2005 (n = 25) and in 10 fast food restaurants or gas station convenience stores in 2006 (n = 43) and 2007 (n = 43). One hundred milliliters of each soft drink was filtered through a hydrophobic grid membrane filter. The remaining portion of the soft drink was kept at room temperature for 4 h before sampling in order to mimic the possible holding time between purchase and consumption. The membrane filters were sampled for total aerobic bacteria, Enterobacteriaceae, lactic acid bacteria, and yeasts and molds. The microbial counts in the 2006 samples were numerically higher than the counts in the 2007 samples except for the average lactic acid bacteria counts, and were either significantly or numerically higher than the counts in the 2005 samples. Soft drinks sampled after the 4-h holding period had relatively higher counts than those sampled initially, with a few exceptions. Some soft drinks had over 4 log CFU/100 ml of total aerobic bacteria, Enterobacteriaceae, lactic acid bacteria, and yeast and mold cells. The study revealed the microbial quality of soft drinks served by dispensing machines in Griffin, GA, and surrounding areas, emphasizing the importance of effective sanitizing practice in retail settings. PMID- 20003748 TI - Bactericidal activity of glutaraldehyde-like compounds from olive products. AB - The bactericidal effects of several olive compounds (nonenal, oleuropein, tyrosol, the dialdehydic form of decarboxymethyl elenolic acid either free [EDA] or linked to tyrosol [TyEDA] or to hydroxytyrosol [HyEDA]), other food phenolic compounds (catechin, epicatechin, eugenol, thymol, carvacrol, and carnosic acid), and commercial disinfectants (glutaraldehyde [GTA] and ortho-phthalaldehyde [OPA]), were tested against strains of Pseudomonas fluorescens, Staphylococcus aureus, Enterococcus faecalis, and Escherichia coli. It was found that the bactericidal activities of olive GTA-like compounds (EDA, HyEDA, and TyEDA) were greater than those exerted by several food phenolic substances. Surprisingly, these olive antimicrobials were as active as the synthetic biocides GTA and OPA against the four bacteria studied. Thus, it has been proposed that the bactericidal activity of the main olive antimicrobials is primarily due to their dialdehydic structure, which is similar to that of the commercial biocides GTA and OPA. Our results clearly reveal that olive GTA-like compounds possess a strong bactericidal activity even greater than that of other food phenolic compounds or synthetic biocides. PMID- 20003749 TI - A foodgrade preparation of nisin from diluted milk fermented with Lactococcus lactis W8. AB - Lactococcus lactis strain W8, which contains the nisin Z gene in its genome, grew well and produced nisin in cow's milk at temperatures of 30 to 37 degrees C. Maximum production of nisin was achieved at 6 h and was 4,000 activity units (AU) per ml in skim milk and 2,400 AU/ml in 3% fat milk. The organism produced nisin even in 20 times diluted skim milk and 3% fat milk at 1,000 and 600 AU/ml, respectively. Boiling of the fermented milk (pH 4.2) made with this culture allowed the separation of the liquid part (whey) from the curd. When 20 times diluted skim milk was fermented and the whey derived from it was lyophilized, the yield of nisin was 60,000 AU/g. The antimicrobial activity of the nisin preparation was stable for at least 1 year at refrigeration temperature. L. lactis W8 may have significant applications in the food industry for a cost effective natural nisin preparation. PMID- 20003750 TI - Modeling Penicillium expansum resistance to thermal and chlorine treatments. AB - Apples and apple products are excellent substrates for Penicillium expansum to produce patulin. In an attempt to avoid excessive levels of patulin, limiting or reducing P. expansum contamination levels on apples designated for storage in packinghouses and/or during apple juice processing is critical. The aim of this work was (i) to determine the thermal resistance of P. expansum spores in apple juice, comparing the abilities of the Bigelow and Weibull models to describe the survival curves and (ii) to determine the inactivation of P. expansum spores in aqueous chlorine solutions at varying concentrations of chlorine solutions, comparing the abilities of the biphasic and Weibull models to fit the survival curves. The results showed that the Bigelow and Weibull models were similar for describing the heat inactivation data, because the survival curves were almost linear. In this case, the concept of D- and z-values could be used, and the D values obtained were 10.68, 6.64, 3.32, 1.14, and 0.61 min at 50, 52, 54, 56, and 60 degrees C, respectively, while the z-value was determined to be 7.57 degrees C. For the chlorine treatments, although the biphasic model gave a slightly superior performance, the Weibull model was selected, considering the parsimony principle, because it has fewer parameters than the biphasic model has. In conclusion, the typical pasteurization regimen used for refrigerated apple juice (71 degrees C for 6 s) is capable of achieving a 6-log reduction of P. expansum spores. PMID- 20003751 TI - Effect of chitosan on the infectivity of murine norovirus, feline calicivirus, and bacteriophage MS2. AB - Chitosan is known to inhibit microorganisms of concern to plants, animals, and humans. However, the effect of chitosan on human enteric viruses of public health concern has not been extensively investigated. The purpose of this study was to determine the effect of chitosan on three human enteric viral surrogates: murine norovirus 1 (MNV-1), feline calicivirus F-9 (FCV-F9), and (ssRNA) bacteriophage MS2 (MS2). Chitosan oligosaccharide lactate (molecular weight of 5,000) and water soluble chitosan (molecular weight of 53,000) at concentrations of 1.4, 0.7, and 0.35% were incubated at 37 degrees C for 3 h with equal volumes of each virus at high (approximately 7 log PFU/ml) and low (approximately 5 log PFU/ml) titers. Chitosan effects on each treated virus were evaluated with standardized plaque assays in comparison to untreated virus controls. The water-soluble chitosan at 0.7% decreased the FCV-F9 titer by approximately 2.83 log PFU/ml, with decreasing effects at lower concentrations, and also decreased MS2 at high titers by approximately 1.18 to 1.41 log PFU/ml, regardless of the concentration used. Chitosan treatments at the concentrations studied had no effect on MNV-1 at high titers. Chitosan oligosaccharide showed similar trends against the viruses, but to a lesser extent compared with that of water-soluble chitosan. When lower virus titers (approximately 5 log PFU/ml) were used, plaque reduction was observed for FCV-F9 and MS2, but not MNV-1. The use of higher-molecular-weight chitosan and at higher concentrations with longer incubation may be necessary to inactivate MNV 1. These results in the plaque reduction of human enteric virus surrogates by chitosan treatment show promise for its potential application in the food environment. PMID- 20003752 TI - Food safety in free-range and organic livestock systems: risk management and responsibility. AB - Animal production systems that offer outdoor access to the animals have become increasingly popular in the Western world due to the growing general discontent of consumers with conventional bioindustrial farming practices. These open production systems offer improved animal welfare but may create new problems for animal health, resulting in increased food safety risks from bacterial, viral, or parasitic infections or environmental contaminants. Examples of these new problems include increased Toxoplasma gondii infections in pigs and high dioxin levels in eggs from free-range hens. In this review, the relation between positive and negative points of free-range and organic livestock production systems is discussed with reference to production in The Netherlands. We investigated how proponents of more animal welfare friendly systems deal with potential negative issues in public and whether any risk communication is used. Generally, we found that the existence of a dilemma is disputed or avoided in communication with the consumer. This avoidance could be detrimental for public trust in alternative animal production systems, should problems occur. To prevent future problems, it will be necessary to communicate about the relevant types and sources of the food safety risks to the consumers. The responsibility for protecting food safety should be properly divided among the various parties involved: producers, processors, governments, nongovernmental organizations, and consumers. PMID- 20003753 TI - Multi-tools approach for food safety risk management of steam meals. AB - Steamed meals comprise a new type of meal in which various raw ingredients are packed together and then cooked by the consumer just before consumption. The presence of raw ingredients and the absence of any inactivation step before home cooking could significantly impact the safety of these meals. In this study some of the many tools available to assess food safety were combined to determine the factors affecting the food safety of this new kind of meal. First the hazards were identified using Stepwise and Interactive Evaluation of Food Safety by an Expert System (SIEFE); then food safety objectives for various food pathogens were determined, and hazard analysis and critical control point (HACCP), Risk Ranger, and predictive microbiology (gamma model) were used to determine the appropriate measures to meet the target set. Finally, links to the performance objective of the cooking stage are also proposed for Salmonella as it had the lowest food safety objectives. The SIEFE methodology excluded only Clostridium botulinum from the possible foodborne pathogens capable of causing foodborne illnesses from these meals, while use of HACCP and modelling demonstrated that cooking is the critical step in preparation of the meals. Risk Ranger was used to rank the possible pathogens: Salmonella and Campylobacter scores were the highest, Bacillus cereus the lowest. Risk Ranger was also used to assess the effect of the cooking stage on food safety and confirmed the importance of this process. PMID- 20003754 TI - Variation in urban and rural water fluoride levels in Ontario. AB - OBJECTIVE: To compare levels of water fluoridation in urban and rural distribution systems in Ontario. METHODS: A random sample of 17 urban and 17 rural municipalities was taken from a list of 445 municipalities. The Ontario Ministry of the Environment (MOE) website was used to identify the water treatment plants that supply these municipalities, and water quality reports published by each of these distribution systems for 2007 were collected. For municipal distribution systems without published reports, staff were contacted directly. RESULTS: Far more urban distribution systems (82%) fluoridate their water compared with rural systems (18%). Most urban water suppliers (14 of 17) meet the 2000 MOE fluoride level standard of 0.5-0.8 mg/L, a range that includes the recently adopted Health Canada standard of 0.7 +/- 0.1 mg/L. Only 3 of 17 rural distributors artificially fluoridate their water and 11 of 16 supply suboptimal levels of fluoride. CONCLUSION: Most Ontarians who live in rural areas receive levels of fluoride that are outside MOE standards. Urban water distribution systems that regulate their fluoride content are compliant with the range recommended in 2000. The communal water supplies of some rural residents of southwestern Ontario contain levels of natural fluoride that are well above the standard for artificially fluoridated water. PMID- 20003755 TI - Use of bispectral index system (BIS) to Monitor Enteral Conscious (moderate) sedation during general dental procedures. AB - Although dental board regulations for the provision of in-office enteral conscious (oral) sedation vary widely with respect to training and pharmacologic strategies, they agree on the use of drugs that are inherently safe, the use of pulse oximetry and the availability of emergency equipment, including pharmacologic antagonists. Patient safety is of greatest concern and is best addressed by appropriate selection of patients, adequate training of personnel and appropriate monitoring of patients. Readings from bispectral index system (BIS) monitors, which use electroencephalographic signals, correlate accurately with depth of sedation during nondissociative general anesthesia of adults and children in the operating room setting. The usefulness of such monitoring as an adjunct to other forms of monitoring of in-office enteral sedation in the dental setting may represent the next important application of this tool, adding a further level of safety for the patient and another level of predictability for the practitioner. This paper reviews the current evidence supporting this new technique, presenting data from 20 procedures in which BIS monitoring during in office enteral sedation was employed in a community dental practice. PMID- 20003756 TI - Radiographic diagnosis of synovial chondromatosis of the temporomandibular joint: a case report. AB - Synovial chondromatosis is a rare, benign condition that usually affects the larger diarthroidal joints of the axial skeleton. Approximately 120 cases of synovial chondromatosis involving the temporomandibular joint (TMJ) have been reported. People with this condition may present with swelling, pain, intracapsular sounds and limitation of mandibular movement. Because it is important to differentiate synovial chondromatosis from other joint pathologies, a thorough history and appreciation of clinical features of these conditions are necessary. Radiographs are an important component of the diagnostic armamentarium for discerning conditions that mimic synovial chondromatosis. A case of synovial chondromatosis diagnosed radiographically with the aid of volumetric computed tomography is described, followed by a discussion of potential causative factors and management strategies. A brief review of the differential diagnosis of synovial chondromatosis involving the TMJ is also provided. PMID- 20003757 TI - E-cadherin protects primary hepatocyte spheroids from cell death by a caspase independent mechanism. AB - Cultivation of primary hepatocytes as spheroids creates an efficient three dimensional model system for hepatic studies in vitro and as a cell source for a spheroid reservoir bioartificial liver. The mechanism of spheroid formation is poorly understood, as is an explanation for why normal, anchorage-dependent hepatocytes remain viable and do not undergo detachment-induced apoptosis, known as anoikis, when placed in suspension spheroid culture. The purpose of this study was to investigate the role of E-cadherin, a calcium-dependent cell adhesion molecule, in the formation and maintenance of hepatocyte spheroids. Hepatocyte spheroids were formed by a novel rocker technique and cultured in suspension for up to 24 h. The dependence of spheroid formation on E-cadherin and calcium was established using an E-cadherin blocking antibody and a calcium chelator. We found that inhibiting E-cadherin prevented cell-cell attachment and spheroid formation, and, surprisingly, E-cadherin inhibition led to hepatocyte death through a caspase-independent mechanism. In conclusion, E-cadherin is required for hepatocyte spheroid formation and may be responsible for protecting hepatocytes from a novel form of caspase-independent cell death. PMID- 20003758 TI - Long-term metabolic and hormonal effects of exenatide on islet transplant recipients with allograft dysfunction. AB - The initial success of islet transplantation (ITx) is followed by graft dysfunction (GDF) and insulin reintroduction. Exenatide, a GLP-1 agonist, increases insulin and decreases glucagon secretion and has potential for beta cell regeneration. To improve functional islet mass, exenatide treatment was given to ITx recipients with GDF. The objective of this study was to assess metabolic and hormonal effects of exenatide in GDF. In this prospective, single arm, nonrandomized study, 11 type 1 diabetes recipients of ITx with GDF had HbA1c, weight, insulin requirements, and 5-h mixed meal tolerance test (MMTT; with/without exenatide given before test) at baseline, 3, 6, and 12 months after initiating exenatide treatment. Baseline MMTT showed postprandial hyperglycemia and hyperglucagonemia. Daily exenatide treatment resulted in improved glucose, increased amylin/insulin ratio, and decreased proinsulin/insulin ratio as assessed by MMTT. Glucagon responses remained unchanged. Exenatide administration 1 h before MMTT showed decreased glucagon and glucose at 0 min and attenuation in their postprandial rise. Time-to-peak glucose was delayed, followed by insulin, proinsulin, amylin, and C-peptide, indicating glucose-driven insulin secretion. Five subjects completed 12-month follow-up. Glucose and glucagon suppression responses after MMTT with exenatide were no longer observed. Retrospective 3 month analysis of these subjects revealed higher and sustained glucagon levels that did not suppress as profoundly with exenatide administration, associated with higher glucose levels and increased C-peptide responses. In conclusion, Exenatide suppresses the abnormal postprandial hyperglucagonemia and hyperglycemia observed in GDF. Changes in amylin and proinsulin secretion may reflect more efficient insulin processing. Different degrees of responsiveness to exenatide were identified. These may help guide the clinical management of ITx recipients. PMID- 20003759 TI - Ambient pollutants as adjuvant for allergic sensitization: the emerging role of platinum group elements. AB - The prevalence of asthma and allergies often observed in urban metropolitan areas as compared to rural and farm communities is still an enigma. Westernized life styles, type of farming and exposure to environmental pollutants seem to simultaneously interact in the determination of this phenotype in genetically predisposed individuals. In this scenario, we asked whether and to what extent we could single out antropogenic airborne contaminants in general, and platinum group elements in particular as relevant causal factors in the generation and in the clinical expression of allergic immune responses in exposed individuals. To this aim, we evaluated epidemiological and basic immunology studies published on peer-reviewed journals indexed in Medline on this subject. We reviewed studies focused on effect of the exposure to platinum group elements on the allergic immune response, with specific reference to our own studies, on their influence on dendritic cells and on the consequent skewing of T-helper and T-regulatory lymphocyte functions. Our laboratory contributed to generate consistent evidence supporting the notion that anthropogenic emissions in general, and platinum group elements in particular, can functionally modulate the immune response in a coordinated pro-allergic fashion. We conclude that in genetically predisposed individuals platinum group elements exert an adjuvant effect specifically leading to more severe allergic reactions. PMID- 20003760 TI - Acute mercury intoxication and use of chelating agents. AB - There is a great hazard of mercury intoxication in the third world for artisanal miners using mercury as amalgam for extracting and refining gold. In developing countries, there is the possibility of risk regarding exposure to Hg from amalgam tooth fillings, ethyl-Hg (thimerosal) added as antiseptic to vaccines and methyl Hg in fish. In one case, a 41-year-old man attempted suicide by ingesting 100 mg of HgCl2. After 8 hours, he developed hematemesis and entered the intensive care unit; his urinary Hg was 10.1 mg/l. Treatment with 2,3-dimercaptopropanol (BAL) was started by intramuscular route after 16 hours at the dosage of 5 mg/kg body weight every 4 hours on days 2-3 and 3 mg/kg every 6 hours on days 4-5 and then every 12 hours on days 6-14 without adverse side effects. Acute Hg intoxication can be managed with BAL as first choice chelator, whereas the less toxic 2,3 dimercaptosuccinic acid (DMSA) and 2,3-dimercaptopropane-1-sulfonic acid (DMPS) should be reserved for cases of less severe inorganic Hg or methyl-Hg acute intoxication. Such agents, recommended only for the treatment of acute Hg poisoning, should not be used for patients suffering from neurological diseases in which environmental Hg exposure is hypothesised. PMID- 20003761 TI - Recurrence of pleomorphic adenoma of the palate after sixteen years: case report and an analysis of the literature. AB - Pleomorphic adenoma or mixed tumour (MT) is a benign neoplasia of slow growth and epithelial histogenesis. We report a particular case of recurring MT of the hard palate. A 39-year-old man came to us with a swelling of the hard half-palate. The patient, 19 years earlier, had had a small formation in the same place that, over a period of three years had slowly grown. Histology showed that it was an MT and it was promptly removed. Sixteen years after the operation, a small recurrence reappeared, reaching a diameter of 12 mm. The patient underwent a new excision. The case reported is of particular interest due to many aspects: the outbreak from the minor salivary glands; the male sex; the young age of the patient at the first sign of the tumour; the appearance of a recurrence after 16 years, not contemplated in literature; and finally, the rapid growth of the second appearance. PMID- 20003762 TI - Infections and mast cells. AB - Mast cells play a role in various physiological functions: innate and acquired immunity, epithelium remodelling and proliferation, angiogenesis, cancer, inflammation and infections. Mast cells are activated by cross-linking of FcERI molecules, which are involved in the binding of multivalent antigens to the attached IgE molecules, resulting in a variety of responses including the immediate release of potent inflammatory mediators. In addition, mast cell biology consists in the capability to secrete preformed mediators which include biogenic amines and newly synthetized mediators, which include lipid-derived mediators and cytokines. It has been reported that parasite infections induce a systemic immunomodulatory network, including regulatory T cells, pro-inflammatory and anti-inflammatory cytokines, which might play a key role in the allergic phenotype. Here, in this article, we revisited the relationship between mast cells and infections. PMID- 20003763 TI - Apoptotic effect of cyclosporin a and dexamethasone in malignant cells of patients with B-chronic lymphocytic leukemia. AB - B-chronic lymphocytic leukemia (B-CLL) is a malignant disorder characterized by the accumulation of the leukemic cells in the G0-G1 phase of the cell cycle and expressing high levels of the anti-apoptotic protein Bcl-2. Since we observed that the treatment of autoimmune complications with Cyclosporine A (CsA) determined in some CLL patients an improvement not only of the autoimmune phenomena, but also of the leukemic process, we evaluated the in vitro cytotoxicity of CsA as compared to Dexamethasone (Dex) on leukemic cells. Leukemic cells obtained from 32 B-CLL patients showed a heterogeneous pattern of spontaneous apoptosis at 24 h interval and this pattern permitted to identify: Group 1 (14/32) with high (>20%) apoptotic rate and Group 2 (18/32) with low cell death. CsA and Dex increased cell death in both groups with a different timing by an apoptotic mechanism that does not involve Bcl-2. Furthermore, in Group 2, CsA induced apoptosis was significant higher than that observed with Dex both at 4 and 24 h. We suggest that, in B-CLL, CsA has a significant pro-apoptotic activity manifested also in patients with low spontaneous apoptosis. Our observations might be taken into account to consider new therapeutic strategies in B-CLL. PMID- 20003764 TI - IL-18 and interferon-gamma in HCV-related hepatocellular carcinoma: a model of interplay between immune status and cancer. AB - Hepatocellular carcinoma (HCC) is a frequent malignancy with a high rate of mortality, and the hepatitis B and C viruses are considered major etiological factors associated with the development of chronic inflammation. Today, there is increasing evidence that the inflammatory process, mediated by the complex cytokine network, is inherently associated with many cancer types, including HCC. In this study we have assayed Th1 cytokines, such as IL-18 and IFN gamma, in the sera of 23 HCC patients with HCV infection, analysing their possible association with HCC in respect to 20 patients: 12 carriers for HCV infection and 8 healthy controls. We have also evaluated the possible difference on IL-18 and IFN gamma in HCC patients with respect to the number of hepatic nodules and rate of tumor differentiation. The mean values of serum IL-18 levels were significantly higher in HCC patients than in HCV carriers (p < 0.001) while IFN gamma serum levels were similar in cases and controls. No significant correlation was present between IL-18 and IFN gamma. In addition, IL-18 was higher in HCC patients with two or more nodules in respect to HCC patients with one nodule (372+/-140 vs 109+/-73 pg /mL; p <0.001). There is no significant difference in HCC patients and no correlation between the cytokines and other evaluated variables such as HCV RNA, alpha-1 fetoprotein, genotype and demographics of HCC patients. Taken together, our data suggest that IL-18 may play a key role in the pathogenesis of HCC and its levels can be utilized as a possible marker in the diagnosis of HCC. PMID- 20003765 TI - Immunohistochemical and transcriptional expression of the matrix metalloproteinases MMP-2 and MMP-9 in normal and pathological human oral mucosa. AB - The oral cavity is exposed to chronic or recurrent, physical and chemical trauma that could lead to mucosal reactions (e.g. hyperplasia, dysplasia and tumors). The objective of this study is to investigate the expression and the possible changes of the two matrix metalloproteinases MMP-2 and MMP-9 in normal and pathological human oral mucosa samples. Normal oral mucosa samples and three different types of pathological conditions (hyperplasia, dysplasia and carcinoma) were used for this study. Immunohistochemical analysis was used to evaluate protein expression for the two enzymes, while Reverse Transcription ? Polymerase Chain Reaction (RT-PCR) was used to investigate gene expression. Image analysis was used to give a quantitative evaluation of the immunohistochemical data. In control samples we identified a weak expression of both MMP-2 and MMP-9 in the epithelial layers. In hyperplasia samples MMPs expression is limited to epithelial layers but the immunoreactivity is more intense than in the control. In dysplasia and carcinoma samples the two matrix metalloproteases are expressed not only in epithelium but also in some cells of the connective tissue and in the vessel walls. Qualitative RT-PCR and image analysis confirmed the immunohistochemical data. The results obtained in this study suggest the existence of a possible relationship between the entity of morphological disorganization of the oral mucosa in different pathologies and the increase of MMP-2 and MMP-9 expression. PMID- 20003766 TI - Severe intravascular haemolysis as delayed manifestation of perivalvular leak in patients with mitral valve replacement: a report of two cases. AB - Haemolytic anaemia following mitral valve replacement is uncommon, however in patients who suffer from some degree of perivalvular leak, severe and potentially fatal recurrent intravascular haemolysis can be an annoying problem. We report the cases of two patients with severe haemolytic anaemia observed some years after mitral valve replacement. In one of the two patients the presence of an association between a valvular leak after mitral valve replacement and a calcific atrial wall produced severe and recurrent haemolysis that required multiple blood transfusions. In the second patient the presence of a single valvular leak after mitral valve replacement induced an episode of haemolytic anaemia some years after the operation. These cases point out that in case of unexplained worsening anaemia, a transthoracic (TT) and transesophageal (TE) echocardiogram should be performed, and the possibility of atrial wall alterations in the producing of anaemia should be kept in consideration. In these cases reoperation resolved the recurrence of anemization. PMID- 20003767 TI - Upper extremity deep vein thrombosis as an uncommon complication of pacemaker implantation: a case report. AB - Venous complications of pacemaker implantation rarely cause immediate clinical problems. An 89-year-old man, without thrombophilia, 4 weeks after a pacemaker implantation experienced functional impotence of the left arm that appeared warm, reddened, oedematous and painful. Color Doppler Ultrasonography revealed a thrombosis of the axillary vein extended to the proximal third of the ulnar vein. In our opinion, upper extremity deep vein thrombosis (UEDVT) represents an important complication of post-surgical pacemaker implantation that should be suspected early, even without specific symptoms and thrombophilia. PMID- 20003768 TI - Human amniotic fluid stem cells culture onto titanium screws: a new perspective for bone engineering. AB - The use of titanium plates and screws for osteosynthesis is considered to be an effective treatment for different kinds of fractures in orthopedic surgery. The aim of the present study is to test the ability of titanium screws to promote the growth of osteoblasts obtained from human amniotic fluid stem cells (AFS). Osteoblastic differentiation was assessed by RT-PCR of specific markers such as COL1, ONC, OPN, OCN, OPG, BMP-4 and Runx2. Mineralization was demonstrated by the presence of red depositions. Adherent cells were found to cover the whole surface of titanium screw by Scanning Electron Microscopy (SEM). The result indicates the excellent growth of osteoblasts obtained from amniotic fluid on a titanium surface and could represent an important point in view of a possible therapeutic application of AFS cells. PMID- 20003769 TI - Recent developments and needs in materials used for personal protective equipment and their testing. AB - The field of personal protective equipment (PPE) has led to several high technology innovations. Indeed, improved protection against the various possible encountered risks is looked for, in particular at the workplace. This has generated the development of new materials and new manufacturing technologies, as well as the introduction of new applications for existing ones. However, the remaining challenges are numerous. This paper presents some of the new technologies introduced in the field of protective clothing against heat and flames, mechanical risks and chemical aggressors. It also describes new challenges that are currently worked on, in particular the effect of service aging and the need for testing methods that reproduce realuse conditions. Finally, it discusses various existing and potential applications of nanomaterials and smart textiles for PPE. PMID- 20003770 TI - Active thermography in qualitative evaluation of protective materials. AB - This is a study of the possibilities of a qualitative evaluation of protective materials with active thermography. It presents a simulation of a periodic excitation of a multilayer composite material. Tests were conducted with lock-in thermography on Kevlar composite consisting of 16 layers of Kevlar fabric reinforced with formaldehyde resin with implanted delamination defects. Lock-in thermography is a versatile tool for nondestructive evaluation. It is a fast, remote and nondestructive procedure. Hence, it was used to detect delaminations in the composite structure of materials used in the production of components designed for personal protection. This method directly contributes to an improvement in safety. PMID- 20003771 TI - A multidimensional approach to modelling for workplace risk assessment. AB - This paper aims to help enhance the process of risk identification and assessment in small enterprises by facilitating the incorporation of insights from accident, human error and risk perception models. This effort takes place through grouping and classification models of all these aspects according to certain criteria, to fit the proper set of models to each situation. To further facilitate the process, the main guidelines of each model are presented. The whole approach is not a new theoretical model but a simplified presentation to help safety engineers in selecting the proper model for the workplace to better assess its risks. An example of the application of this approach is also presented. PMID- 20003772 TI - Mental symptoms and the use of new technical equipment. AB - The aim of this paper was to present how mental symptoms are connected to the use of desktop, portable or minicomputers (communicators and hand-held computers), mobile phones, and background information such as age and gender in the Finnish working-age population. The study was carried out as a cross-sectional study by posting a questionnaire to 15 000 working-age (18-65) Finns. The mental symptoms of 6 121 respondents were analysed using the model factors age, gender, the use of computers and the use of mobile phones. In all data, the use of desktop computers was related to mental symptoms. However, the results of our data are not highly reliable, because the nonresponse rate was over 50%. Nevertheless, it may be essential to take into account in the future that working with computers can increase workers' mental symptoms, and it is important to observe their mental health. PMID- 20003773 TI - On the sound radiation from a circular hatchway. AB - Low-frequency sound radiation from vibrating plates is a practical problem often found in engineering applications. In this article, the sound radiation from a circular hatchway is examined using a discrete approach based in the acoustic resistance matrix. Since this matrix can be combined with the volume velocity vector on the discretized vibrating circular surface, the sound radiation efficiency can be estimated through matrix approaches. The limitation of the approach is discussed by using benchmark results presented in previous works. The method produces acceptable results in low frequencies when the response of the plate is dominated by one low structural mode. When the response of more than one mode is significant, the method gives good estimation of the total sound power just for frequencies up to the first resonance. However, the method can be applied to complex and irregular vibrating plane surfaces. PMID- 20003774 TI - An examination of shoulder postures and moments of force among different skill levels in the wool harvesting industry. AB - The wool harvesting industry employs workers of varying skill levels that differ in both quality and number of harvested fleeces. As it was unknown how skill affected parameters such as joint posture and loading, the current study comparatively examined 140 wool harvesting workers representing 4 skill levels during wool harvesting competitions. Three-dimensional upper limb postures and peak and cumulative shoulder moments were calculated for each worker. Results indicated that elite wool harvesters, in general, used different shoulder postures to perform the harvesting tasks and were thus exposed to different shoulder moments as compared to the lower skill levels. It is plausible that these adopted postures allow the higher class workers to perform their job with higher quality and greater speed as compared to the lower ranked workers. Posturalbased training may help improve technique in lower ranked workers and enable these workers to achieve higher ranked status. PMID- 20003775 TI - Musculoskeletal problems among workers of an Iranian sugar-producing factory. AB - BACKGROUND: Assessment of the level of exposure to work-related musculoskeletal disorders (WMSDs) risk factors can be an appropriate basis for planning and implementing an interventional ergonomics program in the workplace. This study was conducted among workers of an Iranian sugar-producing factory to determine WMSD prevalence rate among production workers and to assess the level of exposure to WMSD risks. MATERIALS AND METHODS: In total, 116 workers were randomly selected from production workshops and included in the study. The Nordic musculoskeletal questionnaire was used to study prevalence of WMSDs and a quick exposure check (QEC) was used to assess physical exposure to risks. Required data were videotaped. RESULTS: Most workers (87.1%) suffered from some kind of MSD symptoms during the 12 months prior to the study. The highest prevalence was reported in knees (58.6%) and the lower back (54.3%). In 99.1% of the workers, the level exposure to MSD risks established with QEC was high and very high. Awkward postures, manual material handling, and long hours of standing were the major ergonomics problems. CONCLUSION: There was a high rate of WMSDs in this factory. The level of exposure to WMSD risk factors was high and corrective measures for reducing risk level were essential. PMID- 20003776 TI - The relationship between psychosocial work factors, work stress and computer related musculoskeletal discomforts among computer users in Malaysia. AB - Increasing numbers of workers use computer for work. So, especially among office workers, there is a high risk of musculoskeletal discomforts. This study examined the associations among 3 factors, psychosocial work factors, work stress and musculoskeletal discomforts. These associations were examined via a questionnaire survey on 30 office workers (at a university in Malaysia), whose jobs required an extensive use of computers. The questionnaire was distributed and collected daily for 20 days. While the results indicated a significant relationship among psychosocial work factors, work stress and musculoskeletal discomfort, 3 psychosocial work factors were found to be more important than others in both work stress and musculoskeletal discomfort: job demands, negative social interaction and computer-related problems. To further develop study design, it is necessary to investigate industrial and other workers who have experienced musculoskeletal discomforts and work stress. PMID- 20003777 TI - The effect of the use of full body harnesses on their protective properties. AB - A full body harness is a component of personal systems protecting against falls from a height. To ensure users' safety, the harness must retain its protective properties during its whole service period. All the elements of a harness (webbings, threads, metal buckles, etc.) are exposed to destructive factors. Harnesses stored and used for a few years were tested. The paper discusses the most frequent types of damage and their effect on the essential resistance parameters. The effect of atmospheric conditions, sunlight, mechanical damage and dust on the parameters of webbings was tested. Conclusions on the main causes of the loss of the protective properties of harnesses are drawn; periodic checks of the physical conditions and guidelines for estimating acceptable service time are recommended. PMID- 20003778 TI - Convenient evaluation of mental stress with pupil diameter. AB - This study proposes a convenient method of evaluating mental stress. The potential of monitoring changes in pupil diameter as a measure of human reliability while operating human-machine systems was examined experimentally. An experiment was carried out to clarify the relation between changes in pupil diameter and autonomic nervous activity by measuring an electrocardiogram and pupil diameter when 10 subjects completed a time-sensitive task. The electrocardiogram was measured using a multitelemeter system and pupil diameter was measured using an eye-mark recorder. Several relationships between changes in pupil diameter and autonomic nervous activity were revealed and indicated that measurement of pupil diameter was an effective indicator of autonomic nervous activity. Therefore, it may be possible to develop a monitoring system that measures changes in pupil diameter as an indicator of the mental state of workers operating human-machine systems. PMID- 20003779 TI - [Pay attention to prevention and treatment of complications in peri-operative period]. PMID- 20003780 TI - [Summary of national conference on peri-operation problems, prevention and treatment of complications]. PMID- 20003781 TI - [Cesarean scar pregnancy analysis of 42 cases]. AB - OBJECTIVE: To discuss the diagnosis and treatment strategy of cesarean scar pregnancy (CSP). METHODS: Forty-two cases of CSP admitted to Peking University First Hospital from Jan. 2003 to Dec. 2008 were analyzed retrospectively, and the clinical characteristics, pelvic Doppler ultrasonography report, management including methotrexate (MTX) therapy, bilateral uterine artery embolism (UAE), curettage and hysterectomy of these 42 women were reviewed. The percentage of human chorionic gonadochopin (hCG) reduction, menstrual period after the treatment of her CSP and recurrent pregnant outcome were also followed. RESULTS: Among the 42 CSP women, 24 (57%) presented with vaginal bleeding, 3 (7%) with lower abdominal pain. The interval between CSP and the last cesarean scar (CS) was 0.5 - 18.0 years and the average duration of gestation was (60 +/- 18) days. Fourteen cases (33%) were finally diagnosed as CSP in our hospital followed by failure of abortion due to misdiagnosed as early uterine pregnancy in other hospitals; one (2%) was finally diagnosed after hysterectomy due to excessive bleeding during curettage because of suspected hydatidiform mole under color sonography; one was diagnosed by MRI after color sonography; the rest 25 women (60%) were all confirmed by color sonography before management. Treatment of these cases included curettage after UAE (17 cases), curettage after UAE and MTX (13 cases), curettage after MTX only (4 cases), MTX only (4 cases) and others (4 cases). No significant difference was found in the average duration of pregnancy, average reduction of serum level of hCG and the operation time for curettage between the first two management group (P > 0.05). All of the 42 cases fully recovered before discharge and the recovery time of menses was averagely (32 +/- 10) days (10 - 60 days) after curettage and 5 reported reduced menstrual blood volume. Three out of the 42 women were pregnant again during follow-up and 1 delivered by CS at 39 weeks of gestation one year after without any complications or CSP. CONCLUSIONS: CSP is not common and can be easily misdiagnosed and color ultrasound scan is important in its early diagnosis. UAE combined with MTX followed by curettage is an effective treatment of CSP. PMID- 20003782 TI - [Clinical significance of preventive treatment of thrombosis for patients undergoing gynecological surgery with high risk factors]. AB - OBJECTIVE: To explore the clinical significance of preventive treatment of thrombosis for patients undergoing gynecological surgeries with high risk factors. METHODS: Prospectively, randomized and cases controlled study was performed in 143 patients underwent gynecological surgery with high risk factors to explore the prevention effectiveness and safety in morbidity of lower extremity deep venous thrombosis (LDVT) and relative effected factors, who were enrolled to three groups according to different preventive treatment: group using intermittent pneumatic calf compression (IPC) in 47 cases, group using low molecular-weight hepairin (LMWH) in 48 cases and control group in 48 cases with no prevention strategy. RESULTS: The morbidity of LDVT on lower extremity was 6% (6/94) in group IPC and 1% (1/96) in group LMWH, 18% (17/96) in control group, which was obviously reduced in group IPC and group LMWH than that in control group (P < 0.05), while there was no significantly difference between group IPC and group LMWH (P > 0.05). After surgery in 3 or 4 days, the morbidity of LDVT in group IPC was 1/6, zero in group LMWH and 71% (12/17) in control group, there was significantly reduced in group IPC and group LMWH than that in control group (P < 0.05). There was no side-effects in group IPC, only 1 case in group LMWH presented small quantity bleed in vagina remnant, while no incision bleeding and bleed tendency. Single-variate analysis indicated that elder age, abdominal surgeries and malignant tumor were as the independent factors to effect preventive treatment of thrombosis (P < 0.05). CONCLUSION: LDVT in patients underwent gynecological surgeries with high risk factors can be obviously reduced and delayed by preventive treatment, which is no side-effects, while be effected by the factors of elder age, abdominal surgeries and malignant tumor. PMID- 20003783 TI - [Efficacy of complications of different surgical treatments in cervical intraepithelial neoplasia III]. AB - OBJECTIVE: To compare the rate of surgical complications and surgical failure of different surgical treatments in cervical intraepithelial neoplasia (CIN) III. METHODS: From Jan 2002 to Jun 2007, 1256 patients with CIN III confirmed by histopathologic punch biopsy and excision biopsy of cervical specimens were analyzed retrospectively. Among them, 74 patients accepted loop electrosurgical excision procedure (LEEP), 869 patients adopted cold knife conization (CKC), 49 patients received vaginal enlarged amputation of cervix, as well as 264 patients accepted external fascia hysterectomy, Chi-square test was used to compare the rate of surgical efficacy and complications of different surgical treatments. RESULTS: The surgical failure incidence of LEEP, CKC, vaginal enlarged amputation of cervix and external fascia hysterectomy were 4.1% (3/74), 0.2% (2/869), 0 and 0.4% (1/264) respectively, which was significantly higher in LEEP than that in CKC (P = 0.004) and external fascia hysterectomy (P = 0.034), while there was no statistical difference between CKC and external fascia hysterectomy (P = 0.549). The rate of surgical complications of LEEP, CKC, vaginal enlarged amputation of cervix and external fascia hysterectomy were 8.1% (6/74), 6.2% (54/869), 6.1% (3/49) and 2.3% (6/264) respectively, which were not statistically significant (P = 0.067) among them. CONCLUSION: LEEP, CKC, vaginal enlarged amputation of cervix and external fascia hysterectomy are all secure and effective procedures for patients with CIN III, who can take the individual choice depending on their different conditions. PMID- 20003784 TI - [Maternal serum level of adiponectin and expression of adiponectin mRNA in subcutaneous and omental adipose tissues in women with preeclampsia]. AB - OBJECTIVE: To determine the expression of adiponectin mRNA in omental and subcutaneous adipose tissues and serum levels of adiponectin in pregnant women complicated with pre-eclampsia (PE) in order to find out its association with the pathogenesis of PE. METHODS: The expression of adiponectin mRNA in omental and subcutaneous adipose tissue of 49 women with severe PE, which were divided into early-onset group (n = 24, presented at 34 weeks of gestation), and 30 healthy pregnant women (control), who presented to Fujian Maternity and Child Health Hospital for prenatal care from September 2007 to July 2008, were determined by real-time fluorescent quantitative reverse transcription PCR. The maternal serum level of adiponectin was detected by enzyme-linked immunosorbent assay for all subjects. RESULTS: (1) The levels of serum adiponectin in the early-onset and late-onset severe PE groups were (5.9 +/- 2.7) and (7.9 +/- 2.8) mg/L, which were significantly lower than that of the control (11.2 +/- 4.2) mg/L, respectively (P < 0.01), and the level in the early-onset group was lower than in the late-onset group (P < 0.05). (2) Among the early-onset, late-onset PE group and the controls, significant difference was found between any two groups of the three in the adiponectin mRNA expression in omental adipose tissue (0.56 +/- 0.17, 0.69 +/ 0.23 and 1.00 +/- 0.22, P < 0.01), but not in the subcutaneous adipose tissue (0.90 +/- 0.19, 0.93 +/- 0.23 and 1.00 +/- 0.16, P > 0.05). (3) The adiponectin mRNA expressions in the subcutaneous and omental adipose tissue were positively correlated with the maternal serum level of adiponectin in all groups (P < 0.05). (4) The omental adiponectin mRNA levels were negatively correlated with maternal serum triglyceride levels in all of the three groups (r = -0.610, -0.659, -0.446, respectively, and all P < 0.05). Negative correlation was also found between the omental adiponectin mRNA level and the prepregnant body mass index in the control group (r = -0.436, P < 0.05). CONCLUSION: Decreased maternal serum adiponectin level may be involved in the pathogenesis of pre-eclampsia and also may contribute to the reduced expression of adiponectin in the omental adipose tissue. PMID- 20003785 TI - [Influence of various hemostatic methods on ovarian reserve function in women with ovarian endometriotic cyst treated by laparoscopic cystectomy]. AB - OBJECTIVE: To investigate the influence on ovarian reserve function in treatment of ovarian endometriotic cyst by laparoscopic cystectomy with various hemostasis management. METHODS: From September 2007 to August 2008, 61 patients with bilateral ovarian endometriotic cyst in Peking University First Hospital and Anzhen Hospital Affiliated to Capital University of Medicine were treated by laparoscopic cystectomy. Those patients were divided into two groups randomly, which were 28 patients in suture group and 33 patients in electro coagulation during the operation. Blood samples were obtained from the patients before the operation, on the day 2 or 3 of the second menstrual cycle after operation and the first menstrual cycle after 6 months operation. The serum levels of follicle stimulating hormone (FSH), luteinizing hormone (LH), estradiol (E(2)) were tested. At the same time, total antral follicles (F(0)) and mean ovarian stromal peak systolic blood flow velocity (PSV) were detected by transvaginal ultrasonography to evaluate ovarian reserve function. RESULTS: There was no significant difference in patients' clinical characteristics and ovarian reserve function between two groups (P > 0.05). In the suture group, the serum level of FSH were (10.2 +/- 2.0) U/L before operation, (11.0 +/- 2.2) U/L on day 2 or 3 of the second menstrual cycle and (11.5 +/- 2.5) U/L on day 2 or 3 of the first menstrual cycle after 6 months'operation respectively. In comparison of data before operation, it exhibited significant difference (P < 0.05); F(0) were 8.9 +/- 2.6 before operation, 7.8 +/- 2.1 on day 2 or 3 of the second menstrual cycle and 7.6 +/- 2.4 on day 2 or 3 of the first menstrual cycle after 6 months' operation. When compared with data before operation, it showed significant difference (P < 0.05); PSV were (0.104 +/- 0.017) m/s before operation, (0.084 +/ 0.016) m/s on day 2 or 3 of the second menstrual cycle, (0.086 +/- 0.022) m/s on day 2 or 3 of the first menstrual cycle after 6 months' operation, it also showed significant difference between preoperation and postoperation (P < 0.01). In the electro coagulation group, the serum level of E(2) were (166 +/- 19), (196 +/- 57), (205 +/- 65) pmol/L, serum level of FSH were (10.0 +/- 1.5), (12.2 +/- 2.6), (13.4 +/- 4.5) U/L, F(0) were 8.9 +/- 2.0, 6.5 +/- 2.0, 6.2 +/- 2.5 (P < 0.01); PSV were (0.101 +/- 0.016), (0.072 +/- 0.021), (0.067 +/- 0.024) m/s before operation and on day 2 or 3 of the second menstrual cycle after operation and the first menstrual cycle after 6 months operation. They all showed significant difference between preoperation and postoperation. In the second menstrual cycle post operation (on day 2 or 3), the data of F(0) and PSV were statistically different between the two groups (P < 0.05); in the first menstrual cycle 6 months after the operation (on day 2 or 3), The serum level of E(2), F(0) and PSV were statistically different between the two groups (P < 0.05). CONCLUSION: It suggested that ovarian reserve function would be decreased in treatment of bilateral ovarian endometriotic cyst by laparoscopic cystectomy, it was more serious when electro coagulation hemostasis were given. PMID- 20003786 TI - [Expression of occludin and zona occluden-1 and their morphologic changes in vagina mucosal cells in patients undergoing vaginal construction by using sigmoid colon]. AB - OBJECTIVE: In order to evaluate the changes of neovaginal defense function in the sight of physical barrier by investigating the changes in the histological morphology and expression of occludin, zona occluden-1 (ZO-1) in mucosal cells of the neovagina in patients undergoing vaginal construction by sigmoid colon. METHODS: Fifteen cases with neovagina constructed by sigmoid colon matched with 15 cases' vaginal mocusa as control specimens were observed with light microscopic and electron microscopic methods. At the same time, the expression of occludin and ZO-1 were semiquantitatively measured by streptavidin-perosidase (SP) immunohistochemical method. RESULTS: (1) Squamous metaplasia was observed in colonic mucosa from lower one-third of the artificial vagina in 10 cases (67%, 10/15). (2) The intercellular tight junctions of neovaginal mucosa became shorter and wider, the number of them were decreased or disappeared by electron microscopic study. (3) In upper neovagina, the positive rates of occludin and ZO 1 expression in mucosal cells were 80% (12/15) and 73% (11/15), respectively. In lower neovaginal mucosa where squamous metaplasia occurred, the positive rates of two protein expression in cells were both 100% (10/10) and also was 100% (15/15) in control group, which showed significant difference (P < 0.05, P < 0.01). In lower neovaginal mucosa where squamous metaplasia hadn't occurred, the positive expression of two protein in cells (4/5, 2/5) were significantly lower than those in lower neovaginal mucosa where squamous metaplasia had happened 100% (10/10), 100% (10/10) and in control group [100% (15/15), 100% (15/15)] (P < 0.05), while there were no significantly difference in positive expression of two protein in cells between this group and upper neovagina (P > 0.05). CONCLUSIONS: The squamous metaplasia had occurred in lower neovaginal mucosa. The decreased expression of occludin and ZO-1 in upper and lower neovaginal mucosa where squamous metaplasia didn't occurred. However, In lower neovaginal mucosa where squamous metaplasia occurred, the expression of occludin and ZO-1 in cells were similar to those in control group, which might play a role in the defense function of neovaginal mucosa. PMID- 20003787 TI - [Safety of promestriene capsule used in postmenopausal atrophic vaginitis]. AB - OBJECTIVE: To investigate the safety and efficacy of promestriene capsule used in the treatment of postmenopausal atrophic vaginitis. METHODS: Fifty-three women at age of 45 - 75 years (more than one year history of menopause) diagnosed with postmenopausal atrophic vaginitis were enrolled in self-control study. They all had typicalsymptoms of postmenopausal vaginitis. Promestriene was given by continuous therapy for 20 days, then maintenance therapy for for 8 weeks (1 pill two times per week used). The level of follicle stimulation hormone (FSH) and estradiol (E(2)) in serum was and thickness of endometrium were detected before and after treatment. The routine biochemical test was used as index to monitoring the safety. The vaginal mature index (VMI), the atrophic vaginitis evaluating score and vaginal healthy evaluating score were evaluated for therapeutic effect. In the mean time, adverse effect was recorded. RESULTS: (1) SAFETY: during promestriene treatment, no case with adverse effect was observed. Before treatment, the mean level of FSH and E(2) was (71 +/- 3) U/L and (41 +/- 18) pmol/L, the mean thickness of endometrium was (2.4 +/- 0.9) mm. After treatment, the mean level of FSH and E(2) was (67 +/- 22) U/L and (43 +/- 37) pmol/L, the mean thickness of endometrium was (2.5 +/- 1.3) mm. No significant difference was observed (P > 0.05). (2) Therapeutic effect: VMI were 42 +/- 15 before and 54 +/- 8 after treatment. The atrophic vaginitis evaluating score were 3.4 +/- 1.7 before and 1.5 +/- 1.4 after treatment. Vaginal healthy evaluating score were 7.8 +/- 2.4 before and 12.0 +/- 2.4 after treatment. They all showed significant difference (P < 0.01). (3) Adverse effect: six cases with vaginal bleeding, 3 cases with breast nodules and 1 case with cervical polyp was observed, however, it was uncertain whether those events were associated with promestriene use. CONCLUSION: The premestriene capsule was safe and effective in the treatment of postmenopausal atrophic vaginitis. PMID- 20003788 TI - [Efficacy and safety of remifemin compared to tibolone for controlling of perimenopausal symptoms]. AB - OBJECTIVE: To investigate the efficacy and safety of remifemin (isopropanolic extract of cimicifuga racemosa) treating perimenopausal symptoms in comparison of tibolone. METHODS: One hundred and eighty postmenopausal women at range of 40 - 60 years old were enrolled in a multicenter, randomized and double blind study. They were divided into remifemin and tibolone group at ratio 1:1. The therapeutic strategy was remifemin 20 mg bid po for 12 weeks in remifemin group and tibolone 2.5 mg qd po for 12 weeks in tibolone group. To evaluate therapeutic effect, total score of Kupperman menopause index (KMI) was used as the major observed index and single item score of KMI were secondary observed index. Safety warning was determined by laboratory tests and adverse events at timepoint of before, at 4 and 12 weeks treatment. RESULTS: (1) Total score of KMI: it were 24 +/- 5 in remifemin group and 25 +/- 6 in tibolone group before treatment. At timepoint of 4 weeks treatment, it were 11 +/- 6 in remifemin group and 11 +/- 7 in tibolone group. At timepoint of 12 weeks treatment, it were 7 +/- 6 in remifemin group and 6 +/- 5 in tibolone group. Total KMI score between two groups did not show statistical difference at various timepoint (P > 0.05). (2) Single item score of KMI: when compared before, at 4 and 12 weeks treatment, did show remarkable difference (P < 0.05) either in remifemin or in tibolone group. However, those single items of KMI score did not show statistical difference between 4 and 12 weeks timepoint in each treatment group (P > 0.05). (3) Adverse effect: the incidence of adverse effect in remifemin group was significantly lower than that of tibolone group. None case with vaginal bleeding was observed in remifemin group, however, 17 cases with vaginal bleeding occurred in tibolone group (19%, 17/90). The incidence of breast swelling were 16% (14/90) in remifemin group and 36% (32/90) in tibolone group; before treatment, the thickness of endometrium were (2.6 +/- 1.1) mm in remifemin group and (2.8 +/- 1.1) mm in tibolone group; at timepoint of 12 weeks treatment, the thickness of endometrium were (2.9 +/- 1.4) mm in remifemin group and (3.4 +/- 2.0) mm in tibolone group. In comparison of thickness of endometrium before and at 12 weeks treatment, no remarkable changes was observed in remifemin group, however, endometrium displayed significantly thicker in tibolone group. CONCLUSIONS: Our study suggested that remifemin was one effective and safe agent to manage women with climacteric symptom. It has similar therapeutic effect and lower incidence of adverse effect when compared with tibolone. PMID- 20003789 TI - [Establishment of endometriosis diagnostic model using plasma protein profiling]. AB - OBJECTIVE: To establish the diagnostic model for endometriosis by screening the plasma biomarkers of endometriosis using surface enhanced laser desorption/ionization time of flight mass spectrometry (SELDI-TOF-MS)coupled with bioinformatics. METHODS: Plasma samples from 36 patients with endometriosis (endometriosis group) matched with 35 patients with infertility or benign ovarian tumors (control group) before laparoscopy were collected at Peking Union Medical College Hospital from January to October 2007. Plasma protein profiling were detected using SELDI-TOF-MS and protein chip and peak intensities were compared between the two groups. Biomarker Discovery Software was used for data analysis and model was build by classification and regression tree software (CART), sensitivity and specificity of the diagnostic model were verified. RESULTS: There were 14 protein peaks abnormally expressed in endometriosis group compared with those of control group (P < 0.01). The diagnostic model composed of three protein peaks with the molecular weight of 3,956,000, 11,710,000 and 6,986,000 showed a sensitivity of 92% and specificity of 83%. In the blind test the model showed a sensitivity of 88% and specificity of 80%. CONCLUSIONS: SELDI-TOF-MS is a new approach for screening markers of endometriosis. Its clinical value deserves further investigation. PMID- 20003790 TI - [Nerve-sparing radical hysterectomy and radical hysterectomy: a retrospective study]. AB - OBJECTIVE: To compare Piver radical hysterectomy (RH) with nerve-sparing radical hysterectomy (NSRH) for cervical cancer patients in terms of postoperative physiology of pelvic autonomic nerve and perioperative complications. METHODS: Ninety-three consecutive patients with invasive cervical cancer underwent RH (69 cases) or NSRH (24 cases) from March 2005 to March 2006 at Fudan University Cancer Hospital. The postoperative function of bladder, bowel and sexual function and perioperative morbidity were assessed. RESULTS: Compared with patients received RH, patients underwent NSRH presented a significantly prompter recovery of bladder function (8.7 vs. 14.8 days, P < 0.01) and bowel function (2.9 vs. 3.2 days, P < 0.01). However, there were not significant difference in terms of operative time (146.7 vs. 143.3 minutes, P > 0.05), estimated blood loss (441.7 vs. 565.9 ml, P > 0.05) and hospital stay (10.21 vs. 10.19 days, P > 0.05). No positive surgical margin was found in both groups. No surgery complication was found in NSRH group, while there were 1 case presented the infection of lymphocyst and 1 case presented intestinal obstruction in RH group. After following up postoperative 6 months, the patients received NSRH had a higher rate of satisfaction at sex activity than those received RH (29% vs. 9%, P = 0.042). CONCLUSION: NSRH is safe and feasible surgical management for cervical cancer patients, which would improved the physiology of pelvic autonomic nerve postoperatively. PMID- 20003804 TI - Optimization of (131)I doses for the treatment of hyperthyroidism. AB - Several methods can be used to determine the activity of (131)I in the treatment of hyperthyroidism. However, many of them do not consider all the parameters necessary for optimum dose calculation. The relationship between the dose absorbed by the thyroid and the activity administered depends basically on three parameters: organ mass, iodine uptake and effective half-life of iodine in the thyroid. Such parameters should be individually determined for each patient in order to optimize the administered activity. The objective of this work is to develop a methodology for individualized treatment with (131)I in patients with hyperthyroidism of the Grave's Disease. A neck-thyroid phantom developed at the IRD was used to calibrate a scintillation camera and a uptake probe SCT-13004 at the Nuclear Medicine Center of the University Hospital of Rio de Janeiro and a uptake probe SCT-13002, available at the Nuclear Medicine Institute in Goiania. The biokinetic parameters were determined based on measurements performed in eight voluntary patients. It is concluded that the use of the equipment available at the hospital (scintillation camera and uptake probe) has shown to be a suitable and feasible procedure for dose optimization in terms of effectiveness, simplicity and cost. PMID- 20003805 TI - 'Voxelization' of Alderson-Rando phantom for use in numerical dose measuring. AB - The use of computational models of the human body (anthropomorphic phantoms) assists in the estimation of the dose absorbed in organs or tissues of people exposed to sources of radiation which are external or internal to them. Nowadays, more realistic anthropomorphic phantoms are based on volume elements, well-known as voxels, and they are constructed from real images obtained through the scanning of people by Computed Tomography (CT) or Magnetic Resonance Imaging (MRI). The existence of artifacts in original images CT or MRI indicates the necessity of utilizing filtering processes before segmentation with the purpose of eliminating noises, improving contrasts and also to detect contours of organs regions. This study presents the methodology used for the creation of a phantom of voxels from tomographic images of Alderson-Rando (AR) physical phantom and the development of a computational model of exposure formed by phantoms resulting from "voxelization" of AR connected to Monte Carlo EGS4 code, added by algorithms to simulate radioactive sources in internal dose measuring. PMID- 20003806 TI - Whole-body measurements at Ipen, Brazil. AB - The intake of radioactive material by workers can occur in the radiopharmaceuticals production or during the handling of these in the medical fields (nuclear medicine). The workers who work in areas where exposures are significant are routinely monitored to demonstrate that the workers are receiving adequate protection from internal contamination. Direct measurements of whole body and thyroid contents provide an estimate of the activity value of these radionuclides in the potentially exposed workers. The whole-body measurements of the workers, trainees and visitors are routinely performed by the In Vivo Monitoring Laboratory (LMIV) of the Energy and Nuclear Research Institute (IPEN/CNEN-SP). The frequency of measurements is defined by the Radioprotection Service (SRP) and the Dose Calculation Group of IPEN. During the period 2006 2007, 2500 measurements had been carried in workers who develop tasks related to the production of radiopharmaceuticals. The activity value of the radionuclides and the workers' tasks relationship had been evaluated. PMID- 20003807 TI - A software to digital image processing to be used in the voxel phantom development. AB - Anthropomorphic models used in computational dosimetry, also denominated phantoms, are based on digital images recorded from scanning of real people by Computed Tomography (CT) or Magnetic Resonance Imaging (MRI). The voxel phantom construction requests computational processing for transformations of image formats, to compact two-dimensional (2-D) images forming of three-dimensional (3 D) matrices, image sampling and quantization, image enhancement, restoration and segmentation, among others. Hardly the researcher of computational dosimetry will find all these available abilities in single software, and almost always this difficulty presents as a result the decrease of the rhythm of his researches or the use, sometimes inadequate, of alternative tools. The need to integrate the several tasks mentioned above to obtain an image that can be used in an exposure computational model motivated the development of the Digital Image Processing (DIP) software, mainly to solve particular problems in Dissertations and Thesis developed by members of the Grupo de Pesquisa em Dosimetria Numerica (GDN/CNPq). Because of this particular objective, the software uses the Portuguese idiom in their implementations and interfaces. This paper presents the second version of the DIP, whose main changes are the more formal organization on menus and menu items, and menu for digital image segmentation. Currently, the DIP contains the menus Fundamentos, Visualizacoes, Dominio Espacial, Dominio de Frequencias, Segmentacoes and Estudos. Each menu contains items and sub-items with functionalities that, usually, request an image as input and produce an image or an attribute in the output. The DIP reads edits and writes binary files containing the 3-D matrix corresponding to a stack of axial images from a given geometry that can be a human body or other volume of interest. It also can read any type of computational image and to make conversions. When the task involves only an output image, this is saved as a JPEG file in the Windows default; when it involves an image stack, the output binary file is denominated SGI (Simulacoes Graficas Interativas (Interactive Graphic Simulations), an acronym already used in other publications of the GDN/CNPq. PMID- 20003808 TI - The effect of the Rochagan on radiolabeling with (99m)Tc. AB - Radionuclides are used in nuclear medicine by variety of diagnostic procedures. The labeling of red blood cells (RBC) with (99m)Tc is a current method applied in clinical nuclear medicine. Drugs can alter this labeling and modify the disposition of the radiopharmaceuticals. The influence of Rochagan on the labeling of blood constituents with (99m)Tc was reported. Samples of blood were incubated with different concentrations of Rochagan (0%; 6.25%; 12.5%; 25%; 50%; 100%). Stannous chloride and (99m)Tc (3.7MBq/mL) were added. Plasma (P) and (RBC) were isolated and precipitated with thricloroacetic acid 5%. The insoluble (IF) and soluble fractions (SF) were separated. The %ATI in RBC, IF-P and IF-RBC were calculated. The %ATI on RBC decreased significantly (p<0.05) from control to all concentrations of Rochagan, respectively: 90.15 + or - 0.14(control) to 70.80 + or - 4.21; to 64.36 + or - 0.33; to 57.30 + or - 1.56; to 50.28 + or - 2.71; to 42.41 + or - 2.24; on IF-RBC, respectively: 84.70 + or - 0.87(control) to 67.16 + or - 4.38; to 63.63 + or - 2.92; to 59.02 + or - 3.17; to 43.75 + or - 1.00; to 24.15 + or - 0.94 and also on IF-P, respectively: 83.46 + or - 1.09(control) to 50.90 + or - 3.36; to 35.46 + or - 4.13; to 35.78 + or - 2.31; to 28.74 + or - 3.09; to 19.66 + or - 1.34. The analyses were performed by T-Student and Mann Whitney tests, p<0.05. This effect was probably due to products present in Rochagan that may complex with ions or have a direct/indirect effect on intracellular stannous ion concentration. PMID- 20003809 TI - A methodology to evaluate occupational internal exposure to fluorine-18. AB - The objective of this work is to develop procedures for internal monitoring of (18)F to be applied in cases of possible incorporation of fluoride and (18)FDG, using in vivo and in vitro methods of measurements. The Na I (Tl) 8" x 4" scintillation detector installed at IRD-Whole Body Counter was calibrated for measurements with a whole body anthropomorphic phantom, simulating homogeneous distribution of (18)F in the body. The NaI(Tl) 3"x 3" scintillation detector installed at the IRD-Whole Body Counter was calibrated for in vivo measurements with a brain phantom inserted in an artificial skull, simulating (18)FDG incorporation. The HPGe detection system installed at the IRD-Bioassay Laboratory was calibrated for in vitro measurements of urine samples with 1 liter plastic bottles containing a standard liquid source. A methodology for bioassay data interpretation, based on standard ICRP models edited with the software AIDE version 6, was established. It is concluded that in vivo measurements have sufficient sensitivity for monitoring (18)F in the forms of fluoride and (18)FDG. The use of both in vitro and in vivo bioassay data can provide useful information for the interpretation of bioassay data in cases of accidental incorporation in order to identify the chemical form of (18)F incorporated. PMID- 20003810 TI - Caffeic acid phenethyl ester inhibits arterial smooth muscle cell proliferation and migration in vitro and in vivo using a local delivery system. AB - Over the last two decades, significant advances have been made in percutaneous coronary intervention (PCI) for the treatment of atherosclerotic plaques. However, restenosis after PCI still challenges both vascular biologists and interventional cardiologists. In this study, we found that caffeic acid phenethyl ester (CAPE) displayed an inhibitory effect on human coronary smooth muscle cell (HCSMC) growth and migration. Flow cytometry analysis showed that the ratio of S phase increased after exposing cells to CAPE for 48-72 h. Pretreatment of cells with CAPE significantly suppressed Cyclin E, CDK2, Cyclin A, and proliferating cell nuclear antibody expression. We demonstrated that CAPE inhibited AKT 1 and MEK1/2 activation. Using a local infusion system, CAPE was able to regress the intima thickening of the iliac artery in rabbits after balloon injury. The percentage of intimal thickening decreased significantly to 55.0 +/- 0.12 in the group after local CAPE infusion compared to the group after saline infusion (98.3 +/- 0.41%). In conclusion, CAPE can inhibit the proliferation and migration of HCSMCs by inducing cell cycle arrest. Decreased cell cycle genes and associated signaling pathway target gene expression may mediate anti-proliferative and anti migration effects of CAPE. Furthermore, CAPE prevents intima thickening in rabbits after balloon angioplasty. These results indicate that CAPE may have therapeutic relevance for the prevention of restenosis during PCI in the treatment of coronary artery diseases. PMID- 20003811 TI - STAT1/2 is involved in the inhibition of cell growth induced by U0126 in HeLa cells. AB - The mitogen-activated protein kinase (MAPK) signaling cascade plays an important role in cell life. Herein we show that small interfering RNAs targeting MAPK1 can inhibit HeLa cell growth and induce apoptosis along with up-regulation of signal transducers and activator 1 and 2 (STAT1/2). However, across-talk between the ras raf-ERK1/2 signalling cascade and the JAK-STAT pathway remain largely unknown. Using MEK inhibitor U0126 and JAK-2 inhibitor AG490, we analyzed the relationship between ERK1/2 and STAT1/2 in HeLa cells. U0126 inhibited HeLa cell growth, arrested the cell cycle at G1/G0, and induced cell apoptosis, and AG490 partially reversed the effects of U0126. U0126 induced up-regulation of ERK1/2 and down regulation of phosphorylated ERK1/2, increased STAT1 and STAT2 expression in a dose-dependent manner, and activated STAT1/2 via their phosphorylation. AG490 markedly inhibited the phosphorylation of STAT1 and STAT2 and slightly increased that of ERK1/2 inhibited by U0126. We suggest that STAT1/2 is involved in the inhibition of cell growth induced by U0126 in HeLa cells. PMID- 20003812 TI - Risk factors of invasive cervical cancer in Morocco. AB - It is now well established that infection with oncogenic human papillomavirus (HPV) types is the necessary cause of cervical cancer (CC) and its immediate precursor cervical intraepithelial neoplasia 3. However, HPV infection alone may not be sufficient to cause CC, and other exogenous and endogenous factors may exist that, in conjunction with HPV, influence the risk of progression from cervical HPV infection to CC. In this chapter, we review the evidence for the role of parity, oral contraceptive (OC) use, and tobacco smoking in CC. In this study, molecular detection of HPV by PCR using consensus primers MY09/MY11 able to amplify the L1 gene present in all types of HPV from fresh frozen biopsies showed that the virus is present in 88% (99/113) of cases. The viral typing, carried out on HPV positive cases through hybridization using specifics probes MY14, WD74, WD126, MY16, MY70 and MY115 which correspond respectively to HPV 16, 18, 31, 33, 45 and 59 has revealed the prevalence of HPV 16 and 18 respectively with 65% (64/99) and 44.4% (44/99). Several combinations of double and multiple infections were also observed. Another part of this work has been devoted to the study of risk factors associated with the development of cancerous cervical lesions. The study is based on a questionnaire during the sampling and also on data collected from clinical records and records of hospital patients. PMID- 20003813 TI - Coculturing embryonic stem cells with damaged hepatocytes leads to restoration of damage and high frequency of fusion. AB - Controversy surrounds issue of cell fusion as a repair mechanism whereby stem cells regenerate. To identify the ratio of fusion happens between stem cells and damaged cells, hepatic cells were damaged with 200microM H2O2 for 2 hr. Then, mouse ESCs were cocultured with damaged human hepatocytes. Fusion was detected directly by karyotyping after 48hr coculture as well as by Oct4 promoter drove GFP signal. Results showed that average ratio of fusion in undamaged control group was 0.031 per thousand while ratio of fusion in damaged group was 0.357 per thousand, which was 10 times higher than fusion happened in the control group. Meanwhile, GFP signal indicated that fusion induced hepatic cells' Oct-4 reactivation. Fusion derived hybrid cells contained chromosomes from both parental cells. Most of the chromosomes were from damaged human hepatic cells. Activity of damage-related enzymes LDH, SGOT and SGPT were significantly lower at 48hr coculture than at 12hr coculture. Expression of albumin in co-culture system was up-regulated after coculture, which indicated the reparation of damage after coculturing. Also, by applying RT-PCR and immunocytochemistry differentiation status of ES cells were evaluated. It was shown that ES cells differentiated to hepatic lineage cells and expressed hepatic genes and proteins. PMID- 20003814 TI - The influence of exon 7 Phe389Leu polymorphism on P120 catenin interactions with E-cadherin and three-dimensional model rebuilt. AB - The significance of endothelial P120 catenin (P120ctn) activity has been recognized for many years, however it was only recently that the complicated regulation of this constitutively expressed enzyme in endothelial cells was identified. A critical component of the P120ctn regulatory cycle in endothelial cells is its intracellular localization to caveolae. The caveolar coordination of P120ctn, more specifically its interaction with E-cadherin plays a major role in normal endothelial P120ctn activity and vascular bioavailability of nitric oxide. We have recently shown that the presence of P120ctn exon 7 Phe389Leu polymorphism caused diminished shear which was dependent catenin activation, was less extensively associated with caveolae, and had a decreased degree of interaction with E-cadherin. Here, we carried out preliminary investigations to identify possible mechanisms of the genotype-dependent endothelial cell responses we observed in our previous investigations. Through this approach we tested the hypothesis that computer simulations could provide insights regarding the contribution of this single nucleotide polymorphism to regulation of the P120ctn isoform. We observed that in the Phe/Leu and Leu/Leu mutant genotypes, the amount of P120ctn associated with E-cadherin was significantly lower. Additionally, we have shown, using a theoretical computational model, that mutation of an amino acid at position 389 might affect the protein-protein interactions and localization of the P120ctn protein. These alterations might also affect the protein function and explain the enhanced disease risk associated with the presence of Phe389Leu polymorphism in the P120ctn protein. PMID- 20003815 TI - Primary immunodeficiencies: "new" disease in an old country. AB - Primary immunodeficiency disorders (PIDs) are rare inborn errors of the immune system. Patients with PIDs are unique models that exemplify the functional and phenotypic consequences of various immune defects underlying infections, autoimmunity, lymphoproliferation, allergy and cancer. Over 150 PID syndromes were characterized in the past 60 years, with an ever growing list of new entities being discovered. Because of their rarity, multi-center collaboration for pooled data analysis and molecular studies is important to gain meaningful insights into the phenotypic and genetic diversities of PIDs. In this article, we summarize our research findings on PIDs in Chinese population in the past 20 years. Close collaboration among various immunology centers, cross-referrals and systematic data analysis constitute the foundation for research on PIDs. Future directions include establishment of a national PID registry, raising awareness of PIDs and securing sufficient resources for patient care and scientific research. PMID- 20003816 TI - Inhibitory receptors of the immune system: functions and therapeutic implications. AB - The immune system has a remarkable ability to respond to seemingly endless antigens. In essence, a productive immune response takes place along a well defined but treacherous line, that is to effectively eradicate pathogens, and at the same time avoid causing damage to self organs. This type of response is fine tuned, at least in part, by a complex array of pathways that either promote or inhibit the activation of innate and adaptive immune cells. Much effort has been focused on pathways that can support immune activation. In this article, we review specifically pathways that can inhibit immune responses and maintain immune homeostasis, highlighting our recent understanding on the role of inhibitory receptors that selectively engage the self MHC class I molecules and the B7 superfamily members, we also discuss the inhibitory Fc receptors and inhibitory cytokines and how such pathways, either individually or collectively, regulate innate and adaptive immune responses. Finally, we summarize new emerging approaches on how such negative pathways can be therapeutically modulated in various disease settings. PMID- 20003817 TI - Paradoxical roles of IL-4 in tumor immunity. AB - Interleukin (IL)-4 is a crucial cytokine in tumor immunology. In the initial murine experiments, IL-4 exhibited potent anti-tumor ability. Tumors genetically modified to produce IL-4 were rejected, while parental tumors grew progressively. Mice rejected IL-4-producing tumors got long-lasting anti-tumor immunity. The comparative study showed that IL-4 induced the most effective immune response among several cytokines in both prophylactic and therapeutic models. All of these indicate IL-4 has strong potential as a tumor therapy agent. However, contrary evidence indeed exists, and is becoming more and more abundant which shows IL-4 is a tumor-promoting molecule. IL-4 amounts are usually elevated in human cancer patients. IL-4 knockout mice are more resistant to tumor challenge than IL-4 competent mice. Furthermore, tumor cells of various histological origins often express increased levels of IL-4 receptor in comparison to their normal counterparts. By carefully examining presently available data, we found the effects of IL-4 in tumor immunity are closely related to its sources, expressing time and dose, as well as the molecular and cellular environments. In this mini review, we concentrate on illustrating the paradoxical roles and underlying mechanisms of IL-4 in tumor immunity and try to understand how one molecule has opposite effects. PMID- 20003818 TI - Immunotherapy of Epstein-Barr virus associated malignancies using mycobacterial HSP70 and LMP2A356-364 epitope fusion protein. AB - Epstein-Barr virus infection is strongly associated with a number of malignancies. The EBV latent membrane protein 2A has been implicated as one of the most attractive candidates for immunotherapy of related malignancies. In previous studies, the T cell epitopes of LMP2A have been identified systematically. However, the epitope-based vaccine generally meets inefficient immunogenicity when used in vivo directly, which could be overcome by combination with appropriate adjuvants. Heat shock protein is a natural chaperon, which is able to activate the classical major histocompatibility complex class I antigen processing pathway (cross-presentation). In this study, a minigene encoding LMP2A(356-364) (FLYALALLL) was genetically fused to the carboxy-terminal of mycobacterial heat shock protein 70. The epitope fusion protein was expressed and purified, and the cross-presentation of LMP2A(356-364) by monocyte-derived dendritic cells pulsed with the epitope fusion protein was evaluated. Results showed that the epitope fusion protein-pulsed mDCs were much more efficient than the single peptide-pulsed mDCs on CTL activation. Immunization of HLA-A2.1 transgenic mice with MtHsp70-LMP2A(356-364) generated peptide specific CTL more effectively than a single peptide plus incomplete Freund's adjuvant (IFA). Growth of LMP2A expressing B16 melanoma tumor cells was suppressed in the vaccinated groups. Our results suggested that MtHsp70-LMP2A(356-364) fusion protein was more effective than the CD8(+) T cell epitope alone on anti-tumor immunity. As a result, the MtHsp70-LMP2A(356-364) fusion protein is considered to be a promising candidate vaccine for EBV related malignancies. PMID- 20003819 TI - The influence of age and sex on the cell counts of peripheral blood leukocyte subpopulations in Chinese rhesus macaques. AB - Non-human primates such as Chinese rhesus macaques are the favorable models for preclinical study of potential therapeutic drugs, vaccines and mechanisms of human diseases. Little is known about the normal levels of leukocyte subpopulations of Chinese rhesus macaques. To obtain these data, 100 blood samples from Chinese rhesus macaques were collected. The normal range of major leukocyte subpopulations, such as T lymphocytes, B lymphocytes, monocytes, myeloid dendritic cells (mDCs) and plasmacytoid dendritic cells (pDCs), were quantitatively analyzed by flow cytometry through BD trucount tubes. The influence of age and sex on the cell counts of leukocyte subpopulations was analyzed. The counts of CD3(+) T cells, CD3(+)CD4(+) T cells, CD3(+)CD8(+) T cells and B cells decreased with age, but those of monocytes, mDCs and pDCs had no significant correlation with age. Significant differences existed in the cell counts of most leukocyte subpopulations between the male and female groups except pDCs. And the values of the females were higher than those of the males. The study provided basic information about the leukocyte subpopulations of Chinese rhesus macaques, and it may be valuable for immunobiological study of Chinese rhesus macaques. PMID- 20003820 TI - Altered endothelin-1 signaling in production of thromboxane A2 in kupffer cells from bile duct ligated rats. AB - Kupffer cells (KCs), the liver resident macrophages accounting for 80-90% of the total population of fixed tissue macrophages in the body, not only play a key role in host defense via removing particulate materials from the portal circulation, but may also contribute to the pathogenesis of various liver diseases. We have previously demonstrated that KCs play an important role in controlling portal hypertension and hepatocellular injury via releasing thromboxane A2 (TXA2) in early fibrosis induced by one-week bile duct ligation (BDL). Production of TXA2 is controlled by cytosolic phospholipase A2 (cPLA2) that is activated by the interaction of entothelin-1 (ET-1) with its G-protein coupled ET receptor B (ETBR). However, the signaling pathways that contribute to the ET-1-induced activation of cPLA2 and production of TXA2 in KCs in the normal healthy or injured livers are not yet clear, which are investigated in the present study using isolated KCs from one-week BDL or sham rats. The pharmacological inhibition of cPLA2 or chelation of intracellular calcium abrogated the ET-1 induction of TXA2 from KCs. Compared to those from sham rats, KCs from BDL animals displayed a significantly enhanced responsiveness of p38 MAPK to ET-1, increased ETBR and Galphai subunit but decreased Galphaq and Galpha11 expression. Inhibition of ERK1/2 or Gq signaling abrogated significantly the ET-1 induction of TXA2 in sham KCs but only slightly in BDL KCs. In contrast, inhibition of p38 MAPK and Gi signaling markedly attenuated the ET-1 induction of TXA2 in BDL KCs but had no effect in sham KCs. Lastly, inhibition of PLC or PKC abrogated ET-1 induction of TXA2 in KCs from both sham and BDL groups. The hepatic stress (such as BDL) induces significant modifications in the receptor and intermediates of ET-1 signaling in KC and subsequently alters ET-1 signaling mechanisms, particularly a shift from Gq induced signaling to Gi induced signaling, in the activation of cPLA2 and production of TXA2 in response to ET-1. PMID- 20003821 TI - A study on the glycan specificity of natural antibody repertoires in rodents. AB - Inbred strains of mice and rats are widely used in preclinical investigations evaluating the effectiveness of glycan-based biomecines, however, the glycan specificity repertoires of serum Abs in rodents have not been fully characterized. In the present study, serum antibodies in naive mice and rats of different inbred strains were analyzed for specificity against 4 representative carbohydrate structures including PGA (1,4-linked alpha-D-galactopyranosyluronic acids), beta-glucan, mannan and alpha-glucan (dextran). Mannan was not recognized by serum Abs from any of the mouse and rat strains. Serum IgM in naive F344, BN and Lewis rats recognized PGA and beta-glucan and, less strongly, dextran. High titer circulating IgM against PGA were found in mice of BALB/c, C57BL/6, C3H/NeH and BXSB strains. C3H/NeH was the only strain which also produced low titer IgM against beta-glucan and dextran. Age-related production of high titer IgM, IgA and IgG Abs against alpha-glucan was observed in BXSB mice. Intraperitoneal immunization of BALB/c and C57BL/6 mice with beta-glucan elicited strong IgM responses, while immunization with PGA also led to an increase of anti-PGA IgM Ab titers. These results provide useful information on the characteristics of glycan specific natural antibody repertoires in rodents. PMID- 20003822 TI - A pilot study of the therapeutic efficacy and mechanism of artesunate in the MRL/lpr murine model of systemic lupus erythematosus. AB - Recent evidence indicates that artesunate has immunomodulatory properties that might be useful for treating autoimmune disease. In this study, we conducted a pilot study and explored the effect and mechanism of artesunate on the treatment of systemic lupus erythematosus using an MRL/lpr murine model. MRL/lpr mice were divided into control, cyclophosphamide (CTX) and artesunate treatment groups. Blood was collected to measure serum levels of creatinine, antinuclear antibody (ANA) and anti-double-stranded DNA (anti-dsDNA) antibody. Twenty-four-hour urine was collected to measure levels of proteinuria. The concentration of monocyte chemotactic protein-1 (MCP-1) in serum and urine was measured. The expression of MCP-1 in kidney was detected by Western blot and immunohistochemistry assay, respectively. The expression of B cell activating factor (BAFF) in spleen was determined by real time-PCR and immunoblotting. We found that artesunate significantly increased the survival rate, body weight and blood leukocyte counts, and reduced the serum levels of ANA and anti-dsDNA antibody titer, 24 h urinary protein, and serum creatinine. Our results indicated that artesunate could decrease MCP-1, major pro-inflammation cytokine, in serum, urine and kidney. We also found that the level of BAFF, the major B cell activation factor, was decreased in artesunate treated MRL/lpr mice. Its efficacy was comparable with that of CTX in this study. Taken together, we have demonstrated that artesunate can inhibit the progression of disease and reverse the pathologic lesion of lupus nephritis. PMID- 20003823 TI - Mitoxantrone-mediated apoptotic B16-F1 cells induce specific anti-tumor immune response. AB - In the process of cell apoptosis induced by specific reagents, calreticulin (CRT) in endoplasmic reticulum is transferred and coated onto the cell membrane. As a sort of specific ligand, the CRT on the surface of apoptotic cells could mediate recognition and clearance of apoptotic cells by phagocytes. In this research we discovered that mitoxantrone could induce apoptosis of mouse melonoma B16-F1 tumor cells, accompanied by the membrane translocation and coating of CRT. When mitoxantrone-treated B16-F1 cells were used as antigen to inoculate mice, the mice acquired an ability to suppress proliferation of homologous tumor cells. Splenocytes from these mice showed an increased cytolytic effect on homologous B16-F1 cells but no such effect on non-homologous H22 tumor cells. All these results suggested that mitoxantrone-treated apoptotic B16-F1 cells could be used as a sort of cell vaccine to initiate effective anti-tumor immunoresponse in mice. PMID- 20003824 TI - Colour vision testing for diabetic retinopathy: a systematic review of diagnostic accuracy and economic evaluation. AB - OBJECTIVE: To determine the diagnostic performance and cost-effectiveness of colour vision testing (CVT) to identify and monitor the progression of diabetic retinopathy (DR). DATA SOURCES: Major electronic databases including MEDLINE, EMBASE, Cumulative Index to Nursing and Allied Health Literature, and Cochrane Database of Systematic Reviews were searched from inception to September 2008. REVIEW METHODS: A systematic review of the evidence was carried out according to standard methods. An online survey of National Screening Programme for Diabetic Retinopathy (NSPDR) clinical leads and programme managers assessed the diagnostic tools used routinely by local centres and their views on future research priorities. A decision tree and Markov model was developed to estimate the incremental costs and effects of adding CVT to the current NSPDR. RESULTS: In total, 25 studies on CVT met the inclusion criteria for the review, including 18 presenting 2 x 2 diagnostic accuracy data. The quality of studies and reporting was generally poor. Automated or computerised CVTs reported variable sensitivities (63-97%) and specificities (71-95%). One study reported good diagnostic accuracy estimates for computerised CVT plus retinal photography for detection of sight-threatening DR, but it included few cases of retinopathy in total. Results for pseudoisochromatic plates, anomaloscopes and colour arrangement tests were largely inadequate for DR screening, with Youden indices (sensitivity + specificity - 100%) close to zero. No studies were located that addressed patient preferences relating to CVT for DR. Retinal photography is universally employed as the primary method for retinal screening by centres responding to the online survey; none used CVT. The review of the economic evaluation literature found no previous studies describing the cost and effects of any type of CVT. Our economic evaluation suggested that adding CVT to the current national screening programme could be cost-effective if it adequately increases sensitivity and is relatively inexpensive. The deterministic base-case analysis indicated that the cost per quality-adjusted life-year gained may be 6364 pounds and 12,432 pounds for type 1 and type 2 diabetes respectively. However, probabilistic sensitivity analysis highlighted the substantial probability that CVT is not diagnostically accurate enough to be either an effective or a cost-effective addition to current screening methods. The results of the economic model should be treated with caution as the model is based on only one small study. CONCLUSIONS: There is insufficient evidence to support the use of CVT alone, or in combination with retinal photography, as a method for screening for retinopathy in patients with diabetes. Better quality diagnostic accuracy studies directly comparing the incremental value of CVT in addition to retinal photography are needed before drawing conclusions on cost-effectiveness. The most frequently cited preference for future research was the use of optical coherence tomography for the detection of clinically significant macular oedema. PMID- 20003825 TI - CIM: beyond a new start. PMID- 20003826 TI - Effects of survivin interference RNA on non-small cell lung carcinoma. AB - OBJECTIVES: The primary purpose of this study was to investigate the in vitro and in vivo effect of survivin interference RNA (siRNA) on non-small cell lung cancer. METHODS: Lentivirus was used as a vector to transfer siRNA into human lung cancer A549 cells. The proliferation of the cancer cells was assessed by MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] assay. The lentivirus-mediated siRNA was also injected into the transplanted A549 tumor tissues in mice. Tumour growth was assessed after 11 injections over a period of 21 days. RESULTS: Compared with the placebo and the blank lentiviral vector groups, the siRNA treatment group had reduced cell growth rate following 4 days of the treatment (P < 0.01). The average size of the transplanted A549 tumours in the siRNA treatment group (0.75+/-0.16 cm3, n=8) was smaller than in the placebo (2.09+/-0.22 cm3, n=6) or the blank lentivrial vector groups (1.89+/-0.18 cm3, n=6) (P < 0.01). The tumour growth inhibition rate in the siRNA groups was 46.1%. CONCLUSION: Lentivirus-mediated siRNA therapy inhibits the growth of human lung cancer cells in vitro. The siRNA therapy also suppresses the growth of the transplanted lung cancer in mice. PMID- 20003827 TI - In vitro antibacterial activity of some systemic and topical antihistaminic preparations. AB - PURPOSE: In vitro antibacterial activity of topical and systemic antihistaminic preparations containing different active substrates against the standard strains of two bacteria was evaluated. METHODS: Four topical and 3 systemic preparations containing pheniramine maleate, chlorophenoxamine hydrochloride, and diphenhydramine hydrochloride were studied. The antibacterial activities of these preparations against strains of S. aureus (American Type Culture Collection, ATCC 29213) and S. epidermidis (ATCC 25212) were tested using the disc diffusion method. In addition, the Minimal Innhibitory Concentration (MIC) and Minimal Bactericidal Concentration (MBC) of parenteral preparations for these two bacteria were determined. RESULTS: Pheniramine maleate-topical and pheniramine maleate-systemic had no activity against bacteria, but the others showed various rates of activity. Chlorophenoxamine hydrochloride-topical and chlorophenoxamine hydrochloride-systemic were the most effective (P < 0.05). Despite the same active substrate content, diphenhydramine hydrochloride-topical-1 and diphenhydramine hydrochloride-topical-2 yielded different results when they were compared with each other or with the other preparations. Diphenhydramine hydrochloride-topical-2 had a relatively higher rate of activity than diphenhydramine hydrochloride-topical-1. Inhibition zone diameters were 16.9+/ 1.5 mm 12.3+/-0.5 mm for S .aureus, 17.4+/-1.0 mm 0 mm for S .epidermidis respectively (P < 0.05). MIC values of parenteral preparations were equal to or above 125 ?g/ml. CONCLUSION: MIC values of parenteral preparations were higher than their blood levels in clinical use. Thus, effects of parenteral preparations may not have been reflected in routine clinical practice. However, topical forms have antibacterial activity due to additive substrates and the use of high concentration levels at the site of application. Therefore, in selection of topical forms for appropriate cases, these effects should also be taken into consideration. The antibacterial activity of topical antihistaminic preparations may be useful in certain dermatological pathology. PMID- 20003828 TI - Assessment of arterial stiffness affected by atorvastatin in coronary artery disease using pulse wave velocity. AB - PURPOSE: Several studies have shown arterial stiffness changes associated with coronary artery disease (CAD). Recently, statins were reported to improve arterial stiffness. The aim of the study was to evaluate the effect of atorvastatin on arterial stiffness in CAD patients using pulse wave velocity (PWV). METHODS: We evaluated 63 patients with hyperlipemia and CAD. Forty-three patients were given 10mg atorvastatin daily and 20 patients were assigned to a low-fat diet. Carotid-femoral PWV (PWV-CF), carotid-radial PWV (PWV-CR) and carotid-distal PWV (PWV-CD) were measured in all patients. RESULTS: Compared with baseline, PWV-CF, PWV-CR and PWV-CD decreased after therapy in the atorvastatin group (13.22+/-3.39 & 11.85+/-2.87; 11.85+/-2.72 & 10.73+/-2.31; and 11.04+/-1.99 & 10.15+/-1.75, P < 0.05). There was no difference in the control group (13.29+/ 2.89 & 13.93+/-2.89; 11.52+/-2.25 & 12.31+/-2.22; and 10.46+/-1.86 & 11.15+/ 1.85, P > 0.05). After treatment, values of total cholesterol, triglyceride and low density lipoprotein (LDL) cholesterol were reduced in the 2 groups (P < 0.05). CONCLUSIONS: The study demonstrated that atorvastatin can improve arterial stiffness in CAD patients independent of its lipid-lowering properties. PMID- 20003829 TI - Relationship between hemoglobin and CD40 ligand in prediabetes. AB - PURPOSE: Hemoglobin (Hb) regulates the endothelial function by modulating the bio availability of NO at the tissue level. A significant direct relationship is present between the Hb levels and endothelial functions in patients with Type 2 diabetes. Testing whether this association also exists in subjects with prediabetes is important because prediabetes is associated with an increased risk of cardiovascular disease and mortality. Therefore, we investigated the association of Hb both with the classical cardiac risk factors and the markers for endothelial dysfunction and inflammation, in subjects with impaired glucose tolerance (IGT). METHODS: We enrolled 69 normotensive, and cardiovascular events free subjects with IGT (M=40, age=45.50+/-6.8 yr). Plasma insulin, hsCRP, soluble CD40L, vonWillebrand factor, p-selectin levels were measured. The parameters given according to the higher and lower median Hb values of the subjects were compared. RESULTS: Subjects with the higher Hb levels exhibited lower HDL-C (46.68+/-10.8 mg/dl vs 51.5+/-8.9 mg/dl; P=0.04) and higher systolic (122.57+/ 6.2 mmHg vs 116.17+/-7.4 mmHg; p < 0.001) and diastolic (79.14+/-3.73 mmHg vs 75.58+/-6.1 mmHg; P=0.005) blood pressures and sCD40L (7.9+/-3.8 ng/ml vs 6.07+/ 2.1 ng/ml; P=0.02) levels. Hb levels were correlated to the HDL cholesterol, sCD40L, systolic and diastolic blood pressures and waist circumference (r=-0.28, P=0.02; r=0.29, P=0.02; r=0.53, P < 0.001; r=0.41, P=0.001; r=0.42, P < 0.001 respectively). According to the multiple logistic regression analysis, Hb was the determinant of sCD40L levels (beta=0.437, P < 0.001). CONCLUSION: These results indicate that there may be a link with higher Hb values and cardiovascular risk factors in patients with IGT. Further investigation is warranted to understand the clinical implications of these findings in subjects with prediabetes. PMID- 20003830 TI - Intra-strain variations of baroreflex sensitivity in young Wistar-Kyoto rats. AB - PURPOSE: To compare baroreflex sensitivity among conscious rats of the same strain. METHODS: Male WKY rats (eight weeks old) were studied. Cannulas were inserted into the abdominal aortic artery through the right femoral artery to measure mean arterial pressure (MAP) and heart rate (HR). Baroreflex gain was calculated as the ratio between variation of HR in function of the MAP variation (?HR/?MAP) tested with a depressor dose of sodium nitroprusside (SNP, 50 microg/kg, iv) and with a pressor dose of phenylephrine (PE, 8 microg/kg, iv). We divided the rats into four groups: 1) Low bradycardic baroreflex (LB), BG between -1 and -2 bpm/mmHg tested with PE; 2) High bradycardic baroreflex (HB), BG < -2 bpm/mmHg tested with PE; 3) Low tachycardic baroreflex (LT), BG between -1 and -2 bpm/mmHg tested with SNP and; 4) High tachycardic baroreflex (HT), BG < -2 bpm/mmHg tested with SNP. Significant differences were considered for p < 0.05. RESULTS: Approximately 82% of the rats presented reduced bradycardic reflex while 22 showed attenuated tachycardic reflex. No alterations were noted regarding basal MAP and HR, tachycardic and bradycardic peak and HR range. CONCLUSIONS: There was alteration in baroreflex sensitivity among rats of the same strain. Care should be taken when interpreting studies employing WKY as a control for the SHR. PMID- 20003831 TI - DD3(PCA3) gene expression in cancer and prostatic hyperplasia. AB - PURPOSE: DD3(PCA3) is a novel gene with characteristics that indicate its potentially valuable role in early identification and diagnosis of malignancy and highly upregulated in transformed cells in PCa. The aim of this work was to validate and analyze, by real-time Reverse Transcription-Polymerase Chain Reaction (RT-PCR), the expression of the DD3(PCA3)gene in a Mexican population, both in intratumoral tissue with PCa and benign prostatic hyperplasia (BPH). METHODS: Human samples from patients with PCa (40 cases) and benign prostatic hyperplasia (40 cases) were analyzed for the mRNA expression of DD3(PCA3)by RT PCR RESULTS: The GAPDH gene showed better stability with a Pearson correlation of 0.953 (P < 0.007) for the determination of housekeeping gene. DD3(PCA3)gene expression was 29.74 times higher in PCa tissue (P < 0.0001) than in BPH. The gene expression for the PCa and BPH was 1731+/-280 and 58.23+/-9.9 fold, respectively. CONCLUSIONS: Determination of DD3(PCA3)gene expression by RT-PCR could be a potentially tool for the early detection of PCa in clinical specimens. PMID- 20003832 TI - A Canadian multi-centre, open-label long-term study of Pegvisomant treatment in refractory acromegaly. AB - PURPOSE: Acromegaly is a rarely diagnosed condition with potentially serious complications including accelerated heart disease and reduced survival. After a mean interval of nearly 9 years from onset of disease, a significant proportion of patients are diagnosed with invasive adenomas precluding complete surgical resection. Furthermore, strict normalization of the growth hormone (GH) target insulin-like growth factor I (IGF-I) cannot always be achieved by adjunctive medical therapy with somatostatin analogues. Here we report the results of a Canadian multi-centre study open-label, dose-titrated long-term study examining safety and efficacy outcomes of a growth hormone receptor antagonist, pegvisomant in 19 patients with refractory acromegaly. METHODS: Previously pegvisomant treated and treatment-naive refractory acromegalic patients at least 18 yr of age were eligible (n=19). Patients received open-label daily subcutaneous injections of pegvisomant adjusted according to IGF-I levels. Safety and IGF-I levels were assessed every 4 to 6 wk. Baseline and follow-up visits at 3-month intervals also included administration of the Signs and Symptoms of Acromegaly Questionnaire. This study is registered with ClinicalTrials.gov, NCT00151437. RESULTS: We show that, in escalating doses, pegvisomant results in age-adjusted normalization of IGF-I in nearly all such patients. This IGF-I normalization occurred early on and was maintained throughout the study period of 27 months (IGF-I standard deviation score (SDS), mean +/- SE: 1.66 +/- 0.36, P=0.0003 vs baseline), with a nadir at 18 months (IGF-I SDS, mean +/- SE: 1.50 +/- 0.38, P=0.0010 vs baseline). IGF-I control was also accompanied by measurable improvements in disease-associated symptoms and without radiographic evidence of pituitary tumour progression. Overall, the safety profile of pegvisomant therapy in this patient population was found to be satisfactory and suitable for a long-term treatment. CONCLUSION: Our findings provide support for the long-term safety and efficacy of the GH receptor antagonist pegvisomant in achieving IGF-I control in patients with refractory acromegaly. PMID- 20003833 TI - beta-Tubulin III mRNA expression and docetaxel sensitivity in non-small cell lung cancer. AB - PURPOSE: Despite the success of docetaxel as an anti-tumour agent, the inter individual variability in drug response still poses a major impediment to further use of this agent in the treatment of cancer. Current knowledge about predictive biomarkers of docetaxel sensitivity in malignant effusions is poor. The aim of this study was to investigate the association between beta-tubulin III mRNA expression and chemosensitivity to docetaxel in metastatic malignant effusions. METHODS: Real-time quantitative PCR was used for analysis of beta-tubulin III mRNA expression in 37 malignant effusions collected prospectively. Viable tumour cells obtained from malignant effusions were tested for sensitivity to docetaxel using ATP-TCA assay. RESULTS: beta-tubulin III expression was inversely correlated with sensitivity to docetaxel in pleural effusions of NSCLC patients (P =0.022). The lower level of beta-tubulin III mRNA expression in malignant effusions was associated with higher chemosensitivity to docetaxel in NSCLC patients in vitro. No correlation was found between beta-tubulin III mRNA expression and docetaxel sensitivity in malignant effusions of gastric cancer patient. CONCLUSION: Our results demonstrated that beta-tubulin III mRNA expression level in malignant effusions, in which all cancer cells were metastatic, was correlated with docetaxel sensitivity in NSCLC. This highlights the potential role of biomarkers in malignant effusions in further customized chemotherapy. PMID- 20003834 TI - Hypertension and hemostatic/fibrinolytic balance disorders. AB - BACKGROUND: Hypertension is one of the principal risk factors for cardiovascular disease. We aimed to evaluate the impact of hypertension on fibrinolytic balance and endothelial function by measuring plasma levels of plasminogen activator inhibitor-1 (PAI-1), tissue plasminogen activator antigen (tPA), tPA/PAI-1 complex and fibrinogen. METHODS: Patients enrolled into the study were divided into four groups: 22 essential hypertensive (EH), 22 white coat hypertensive (WCH), 22 renovascular hypertensive (RH) and 22 normotensive control subjects. Plasma PAI-1, tPA, tPA/PAI-1 complex levels were measured by enzyme linked immunosorbent assays. RESULTS: There was no difference in the systolic and diastolic blood pressure measurements of the EH and RH groups. The four groups were comparable for age, gender, smoking habits and BMI. Patients with EH, RH and WCH had increased plasma levels of PAI-1, tPA, tPA/PAI-1 complex and fibrinogen compared with controls. No fibrinolytic parameter was associated with blood pressure in hypretensive subjects. CONCLUSION: This prospective study showed that fibrinolytic markers such as PAI-1, tPA, tPA/PAI-1 complex are independently associated with the development of hypertension. This supports the hypothesis that disturbances in fibrinolysis precede a cardiovascular event. Therefore, hypertension may be associated with impaired fibrinolysis. PMID- 20003835 TI - Physiologic hyperleptinemia in obesity does not affect vasopressin secretion in acute hypo- or hyperosmolality. AB - PURPOSE: Abnormal water excretion after ingestion of a water load has been described in obesity. We hypothesized that AVP secretion is abnormal in obese subjects in acute hypo- and hyperosmolality and that the hormone leptin is partly responsible for this. METHODS: We studied the relation between leptin, AVP and serum osmolality in two separate tests: (1) after ingestion of a water load (20 ml/kg lean body mass plus 5 ml /kg of adipose tissue) and (2) after iv hypertonic saline (5% NaCl) at a rate of 0.1 ml/kg lean body mass/minute for 120 min in ten subjects of normal weight (BMI > 20 and < 25 kg/m2; controls) and ten obese females (BMI > 30 kg/m2). Obese subjects were tested before (98.6 +/- 9.3 kg) and after weight loss (90.2 +/- 8.5 kg). RESULTS: In the water load experiment, obese subjects excreted a smaller percentage of the water load than controls. Weight loss restored the ability to excrete the water load in the obese. In the water load and hypertonic saline infusion experiment, plasma AVP concentrations and the area under the curve (AUC) for AVP concentration were not different in obese from normal weight women. Baseline leptin concentration was not correlated with baseline AVP or the change in AVP during the experiment in any of the groups. Weight loss did not change AVP responses in obese subjects. CONCLUSION: AVP secretion in response to acute hypo- and hyperosmolality is not different in normal weight and obese subjects. There is no correlation between leptin and AVP in normal weight or obese subjects. PMID- 20003836 TI - Exercise capacity of children with pediatric lung disease. AB - BACKGROUND: Pulmonary function of children with cystic fibrosis (CF) and bronchopulmonary dysplasia (BPD) is similar at rest even though the mechanisms of injury differ. We sought to compare the peak exercise responses in children with BPD versus CF while controlling for pulmonary impairment, nutritional status, gender, age, height, and predicted forced expired volume in 1 second (approximately 73% of predicted). METHODS: Nine BPD children and 9 CF children underwent spirometry and a progressive exercise test to maximum on a cycle ergometer. RESULTS: There was no difference between groups in body mass percentile (CF:97 +/- 13%, BPD: 98 +/- 11%), peak power output (Wpeak) (CF:67 +/- 19 W, BPD:73 +/- 28 W), % predicted Wpeak (CF:83 +/- 28%, BPD:88 +/- 15%), peak oxygen uptake (VO2peak, CF: 38 +/- 7 ml/kg/min, BPD: 39 +/-6 ml/kg/min), or % predicted VO2peak (CF:99 +/- 16 %, BPD:96 +/- 27%). CONCLUSIONS: Children with mild pulmonary impairments are able to achieve a near normal peak power output and a normal VO2peak. Neither the aetiology nor the developmental onset of the process appears to be important influences on VO2peak or Wpeak. PMID- 20003837 TI - The effect of MTHFR c.677C>T on plasma homocysteine levels depends on health, age and smoking. AB - PURPOSE: The role of homocysteine in the pathogenesis of arteriosclerosis and stroke is under debate. It is important to determine the interplay of factors that influence homocysteine plasma levels, such as age, gender, smoking and the genetic background. The T-allele of the common variant methylenetetrahydrofolate reductase (MTHFR) c.677C > T is the most prevalent known genetic cause of elevated plasma homocysteine levels, but the association of this allele with vascular disease has been controversial. The aim of the present study was to examine whether the influence of MTHFR c.677C > T on homocysteine levels depends on individual factors. METHODS: From an ongoing study on atherosclerosis, we analyzed 523 Caucasian individuals, including patients with cerebrovascular disease (n=141), their healthy spouses (n=106) and the offspring (n=276). ANOVA and regression analyses were employed to separately analyze the effect of MTHFR c.677C > T on homocysteine levels in patients, spouses and offspring as well as in subgroups defined by age, gender and smoking. RESULTS: MTHFR c.677C > T was associated with homocysteine plasma levels in the study sample (P < 0.001), but not in the patients with cerebrovascular disease, if analyzed separately. Analyses of subgroups divided by the age of 55 years revealed that the MTHFR c.677 C > T genotype was associated with homocysteine plasma levels in the younger (P < 0.001), but not in the older individuals. In addition, if individuals with at least less than ten cigarette package years were analyzed separately, MTHFR c.677C > T was associated with plasma homocysteine levels only in the group with the lower cigarette consumption (P=0.002). CONCLUSION: In our study, the association of the MTHFR c.677C > T genotype with plasma homocysteine levels was weakened by other factors that impact homocysteine levels. The effect of MTHFR c.677C > T on plasma homocysteine levels may, thus, be of major importance for healthy, young, non-smoking persons. Such specifications may explain the controversial results of epidemiological studies on the relevance of MTHFR c.677C > T. PMID- 20003838 TI - Prognostic value of CRM1 in pancreas cancer. AB - PURPOSE: Pancreatic cancer is a highly aggressive malignant tumour with poor prognosis. The median survival is only 6 months. This study investigated the prognostic value of nuclear export protein chromosomal region maintenance/exportin 1/Xpo1 (CRM1) expression in pancreas cancer. METHODS: CRM1 expression was detected, by Western blot, in pancreatic tissue from 69 cancer patients and 10 normal subjects. RESULTS: CRM1 showed increased expression in pancreatic cancer tissue (P = 0.007). The high expression of CRM1 was associated with increased serum levels of CEA (P = 0.002) and CA19-9 (P = 0.005). There was an association between CRM1 expression and tumour size (P = 0.01), lymphadenopathy (P = 0.004) and liver metastasis (P = 0.003). High CRM1 expression was not correlated with the other clinicopathological parameters. High CRM1 expression was a prognostic indicator for progression-free survival (PFS) (P = 0.006) as well as overall survival (OS) (P = 0.001). Expression of CRM1 was an independent prognostic parameter for poorer PFS and OS (95% CI, 1.27-5.39). CONCLUSIONS: CRM1 expression demonstrated prognostic value in pancreatic cancer. Prospective studies are required to determine the prognostic role of high expression of CRM1 in pancreatic cancer. PMID- 20003839 TI - Comparison of antihyperglycemic effects of creatine and metformin in type II diabetic patients. AB - PURPOSE: To compare the antihyperglycemic effects of metformin and creatine in recently detected type II diabetics in a short-term clinical study. METHODS: In a 14 day simmetrically randomized crossover study, recently detected type II diabetics received either creatine (2x3 g/day) or metformin (2x500 mg/day) for five days, followed by two days of washout, followed by cross-over to the opposite treatment for the next five days. Fasting and post-prandial (-15, 60, 90, 120, 180 and 240 min) blood glucose, insulin, c-peptide, creatine and lactate were measured every other day for the duration of treatment, and HbA1c only at the begining and at the end of the study. RESULTS: Both creatine and metformin decreased glucose concentrations to similar levels at all time points vs. basal glucose values [-15, 60, 90, 120, 180, and 240 min]: 11.1+/-0.75 vs 9.1+/-0.55a vs 8.8+/-0.59b, 14.4+/-0.6 vs 12.9+/-0.47a vs 13.1+/-0.55a, 14.8+/-0.58 vs 13.0+/ 0.46b vs 13.3+/-0.55a, 14.1+/-0.6 vs 11.9+/-0.42b vs 12.5+/-0.51a, 12.2+/-0.6 vs 9.6+/-0.36c vs 9.9+/-0.38c, and 10.1+/-0.47 vs 7.8+/-0.36c vs 8.4+/-0.4b; (aP < 0.05; bP < 0.01; cP < 0.001 vs. basal glucose values). Neither treatment altered insulin, c-peptide, or HbA1c. Lactate varied during the day, but never reached the upper level of the safety reference range. CONCLUSION: Short-term treatment with creatine and metformin elicits similar glucose lowering effects in recently detected type II diabetics. Further studies are necessary to determine the effect of creatine on long-term glucose and insulin regulation. PMID- 20003840 TI - Coordinated regulation of follicle development by germ and somatic cells. AB - The continuum of folliculogenesis begins in the fetal ovary with the differentiation of the oogonia and their isolation within the primordial follicles. Primordial follicle activation is an enigmatic process, whereby some follicles enter the growing pool to become primary follicles, thereby embarking on an irreversible progression towards ovulation or atresia. This process is under the coordinated regulation of factors from the oocyte itself, as well as from the somatic cells of the ovary, in particular the theca and granulosa cells, which are structural components of the follicle. These two influences provide the principal stimuli for the growth of the follicle to the late preantral or early antral stage of development. The endocrine effects of the gonadotrophins FSH and LH are essential to the continued progression of the follicle and most atresia can be attributed to the failure to receive or process the gonadotrophin signals. The peri-ovulatory state has received intensive investigation recently, demonstrating a coordinated role for gonadotrophins, steroids, epidermal growth factor family proteins and prostaglandins. Thus, a complex programme of coordinated interaction of governing elements from both germ and somatic cell sources is required for successful follicle development. PMID- 20003841 TI - Mammalian oocyte development: checkpoints for competence. AB - During the lifespan of the female, biochemical changes occur in the ovarian environment. These changes are brought about by numerous endogenous and exogenous factors, including husbandry practices, production demands and disease, and can have a profound effect on ovarian oocyte quality and subsequent embryo development. Despite many investigations, there is no consensus regarding the time or period of follicular oocyte development that is particularly sensitive to insult. Here, the key molecular and morphological events that occur during oocyte and follicle growth are reviewed, with a specific focus on identifying critical checkpoints in oocyte development. The secondary follicle stage appears to be a key phase in follicular oocyte development because major events such as activation of the oocyte transcriptome, sequestration of the zona pellucida, establishment of bidirectional communication between the granulosa cells and the oocyte and cortical granule synthesis occur during this period of development. Several months later, the periovulatory period is also characterised by the occurrence of critical events, including appropriate degradation or polyadenylation of mRNA transcripts, resumption of meiosis, spindle formation, chromosome alignment and segregation, and so should also be considered as a potential checkpoint of oocyte development. PMID- 20003842 TI - Is the zona pellucida an efficient barrier to viral infection? AB - Although the transfer of embryos is much less likely to result in disease transmission than the transport of live animals, the sanitary risks associated with embryo transfer continue to be the subject of both scientific investigations and adaptations of national and international legislation. Therefore, the implications are important for veterinary practitioners and livestock breeders. In vivo-derived and in vitro-produced embryos are widely used in cattle and embryos from other species, such as sheep, goats, pigs and horses, are also currently being transferred in fairly significant numbers. Bearing in mind the wide variety of embryos of different species and the correspondingly large number of viruses that are of concern, it is expedient at this time to look again at the importance of the zona pellucida (ZP) as a barrier against viruses and at the susceptibility or otherwise of embryonic cells to viral infection if ever they are exposed. For embryos with an intact ZP, viral infection of the embryo is unlikely to occur. However, the virus may stick to the ZP and, in this case, International Embryo Transfer Society (IETS) washing procedures in combination with trypsin treatment are mandatory. A caveat is the fact that currently more and more types of embryos are becoming available for transfer and scientific data cannot be extrapolated from one species to another. These topics are discussed in the present review. PMID- 20003843 TI - Towards the use of microfluidics for individual embryo culture. AB - Mammalian embryo development is still relatively inefficient in vitro. Much research has been conducted on the chemical environment, or culture medium, surrounding the embryo, but little attention has been given to the actual physical culture environment, which has changed very little over the years. The application of microfluidics to embryo production in vitro is a tantalising approach that may alleviate some of the limits that traditional microdrop culture places on embryo development and research into gamete and embryo physiology. These devices may lead to enhanced in vitro embryo development and quality by more closely mimicking the in vivo environment. Initial work in this area is promising and gives us proof-of-principle that these unique microfluidic systems may indeed be applicable to in vitro culture of gametes and embryos. The present paper reviews the advantages of microfluidics for in vitro embryo production: how the platforms are manufactured, the current uses of microfluidics in assisted reproduction, static v. dynamic culture environments, individual gamete and embryo culture and the future directions of microfluidic application to in vitro embryo production and manipulation. Finally, preliminary data from our laboratory using a new microfluidic well insert for porcine, bovine and murine embryo culture is discussed. PMID- 20003844 TI - Challenge testing of gametes to enhance their viability. AB - Embryos, oocytes and spermatozoa undergo several manipulations during the in vitro procedures that are an integral part of assisted reproductive technologies (ART) in mammals. Consequently, some of the gametes are damaged irreparably, whereas others react to these challenges with some sort of survival mechanism that enables them to come through the process. The details of the mechanism remain unknown but, if identified, it could have immense potential as a new way to improve the viability of embryos produced by ART. However, few publications describe systematic ways to challenge test gametes and then to use the results as a basis for improving gamete viability. Furthermore, new methods to monitor the reactions of gametes to such challenge tests are needed. In the present review, these two issues are discussed, as are some of the conditions necessary before a challenge test protocol can be part of future work with ART. PMID- 20003845 TI - Applications of RNA interference-based gene silencing in animal agriculture. AB - Classical genetic selection, recently aided by genomic selection tools, has been successful in achieving remarkable progress in livestock improvement. However, genetic selection has led to decreased genetic diversity and, in some cases, acquisition of undesirable traits. In order to meet the increased demands of our expanding population, new technologies and practices must be developed that contend with zoonotic and animal disease, environmental impacts of large farming operations and the increased food and fibre production needed to feed and clothe our society. Future increases in productivity may be dependent upon the acquisition of genetic traits not currently encoded by the genomes of animals used in standard agricultural practice, thus making classical genetic selection impossible. Genetic engineering of livestock is commonly used to produce pharmaceuticals or to impart enhanced production characteristics to animals, but has also demonstrated its usefulness in producing animals with disease resistance. However, significant challenges remain because it has been more difficult to produce animals in which specific genes have been removed. It is now possible to modify livestock genomes to block expression of endogenous and exogenous genes (such as those expressed following virus infection). In the present review, we discuss mechanisms of silencing gene expression via the biology of RNA interference (RNAi), the technology of activating the RNAi pathway and the application of this technology to enhance livestock production through increased production efficiency and prevention of disease. An increased demand for sustainable food production is at the forefront of scientific challenges and RNAi technology will undoubtedly play a key role. PMID- 20003846 TI - Effects of gamete source and culture conditions on the competence of in vitro produced embryos for post-transfer survival in cattle. AB - One limitation to the use of in vitro-produced embryos in cattle production systems is the fact that pregnancy rates after transfer to recipients are typically lower than when embryos produced in vivo are transferred. Conceptually, the oocyte and spermatozoon from which the embryo is derived could affect competence for post-transfer survival. There are sire differences in embryonic survival after transfer, but there is little evidence that an embryo's ability to establish pregnancy is determined by sex sorting of spermatozoa by flow cytometry. The role of the source of the oocyte as a determinant of embryonic survival after transfer has not been examined carefully. Conditions for embryo culture after fertilisation can have an impact on the ability of the embryo to establish pregnancy following transfer. Among the specific molecules produced in the reproductive tract of the cow that have been shown to improve competence of in vitro-produced embryos for post-transfer survival are colony-stimulating factor 2, insulin-like growth factor-1 (for recipients exposed to heat stress) and hyaluronan (for less-advanced embryos). There is also a report that embryo competence for post-transfer survival can be improved by inclusion of a carbon activated air filtration system in the incubator used to culture embryos. Progress in developing culture systems to improve embryonic competence for survival after transfer would be hastened by the development of in vitro assays that accurately predict the potential of an embryo to establish pregnancy after transfer. A group of 52 genes has been identified that are differentially expressed in embryos that developed to term v. embryos that did not establish pregnancy. Perhaps a gene microarray consisting of these genes, alone or in combination with other genes, could be used to screen embryos for competence to establish pregnancy. PMID- 20003848 TI - Pregnancy recognition and abnormal offspring syndrome in cattle. AB - Development of the post-hatching conceptus in ruminants involves a period of morphological expansion that is driven by complex interactions between the conceptus and its intrauterine environment. As a result of these interactions, endometrial physiology is altered, leading to establishment of the pregnancy and continued development of the placenta. Disruption of normal fetal and placental development can occur when embryos are exposed to manipulations in vitro or when inappropriate endocrine sequencing occurs in vivo during the pre- and peri implantation periods. The present review addresses the development of the post hatching bovine conceptus, its interactions with the maternal system and changes in development that can occur as a result of in vivo and in vitro manipulations of the bovine embryo. PMID- 20003847 TI - Bovine embryo transfer recipient synchronisation and management in tropical environments. AB - Numerous studies have shown that it is possible to manipulate follicular and luteal dynamics, thereby eliminating the need for oestrus detection in embryo transfer (ET) programmes. Fixed-time ET (FTET) protocols are based on the use of gonadotrophin-releasing hormone (GnRH) and prostaglandin (PG) F or progesterone/progestogen (P4)-releasing devices and oestradiol. The FTET protocols increases the proportion of recipients transferred, and therefore pregnancy rates, compared with the use of PGF followed by ET 7 days after oestrus. Furthermore, the addition of equine chorionic gonadotrophin (eCG) to the P4 and oestradiol-based FTET protocols results in an even higher proportion of recipients transferred, and thus higher pregnancy rates. The beneficial effect of eCG treatment may be related to increased growth of the dominant follicle and increased plasma P4 concentrations during the subsequent luteal phase. In Bos taurus x Bos indicus recipients, pregnancy rates were positively correlated with the diameter of the corpus luteum (CL) and the number of CL at ET. When repeat breeder Holstein cows were used as recipients, FTET protocols increased number of recipients transferred and pregnancy rates compared with the traditional PGF based synchronisation protocols. In conclusion, the use of FTET protocols eliminates the need for the detection of oestrus and results in a greater proportion of recipients transferred and satisfactory pregnancy rates. Thus, FTET optimises the use of recipients, reducing labour and animal handling and facilitating the use of ET. PMID- 20003849 TI - Delivery of cloned offspring: experience in Zebu cattle (Bos indicus). AB - The production of a healthy cloned calf is dependent on a multitude of successful steps, including reprogramming mediated by the oocyte, the development of a functional placenta, adequate maternal-fetal interaction, the establishment of a physiological metabolic setting and the formation of a complete set of well differentiated cells that will eventually result in well-characterised and fully competent tissues and organs. Although the efficiency of nuclear transfer has improved significantly since the first report of a somatic cell nuclear transfer derived animal, there are many descriptions of anomalies concerning cloned calves leading to high perinatal morbidity and mortality. The present article discusses some our experience regarding perinatal and neonatal procedures for cloned Zebu cattle (B. indicus) that has led to improved survival rates in Nellore cloned calves following the application of such 'labour-intensive technology'. PMID- 20003851 TI - New approaches to superovulation in the cow. AB - There is continuing need to simplify bovine superovulation protocols without compromising embryo production. The control of follicular wave emergence and ovulation has facilitated donor management, but the most commonly used treatment, oestradiol, cannot be used in many parts of the world and mechanical removal of the dominant follicle is difficult to apply in the field. Other alternatives include gonadotrophin-releasing hormone (GnRH) or LH, but efficacy in groups of randomly cycling animals is variable. Another alternative is to increase the response to GnRH by inducing a persistent follicle and initiating FSH treatments following GnRH-induced ovulation. The number of transferable embryos following superovulation during the first follicular wave did not differ from that achieved 4 days after oestradiol benzoate and progesterone. To further simplify superovulation, FSH has been administered as a single intramuscular injection. Superovulation of beef donors with a single intramuscular injection of Folltropin V (Bioniche Animal Health, Belleville, ON, Canada) diluted in a slow-release formulation resulted in embryo production comparable to that obtained using the traditional twice-daily protocol. The single intramuscular injection has the potential to reduce labour and handling and may be useful when handling stress is an impediment to success. These alternatives provide ways of facilitating widespread application of embryo transfer technologies. PMID- 20003852 TI - New approaches to superovulation and embryo transfer in small ruminants. AB - The present paper reviews the current state of knowledge in multiple ovulation and embryo transfer (MOET) technology in small ruminants, focusing on recently reported information. Major new findings are related to follicular wave patterns in small ruminants, the elucidation of follicular dominance and the integration of this information into ovarian superstimulatory treatment protocols. Follicular dynamics determine steroid and gonadotrophin secretion, follicular responses to FSH, ovulatory responses and embryo yield. Protocols that control follicular dominance have been designed to allow the initiation of superstimulation at the beginning of a follicular wave. New approaches consist of (1) synchronisation of ovulation for superstimulation during Wave 1 (i.e. Day 0 protocol), (2) pretreatment with a gonadotrophin-releasing hormone (GnRH) antagonist from 10 days prior to FSH treatment to avoid follicular dominance and (3) progesterone oestradiol cotreatment to synchronise follicle wave emergence. These protocols provide a homogeneous pool of small follicles that are gonadotrophin responsive, enhancing the superovulatory response and embryo yield with a reduction in the incidence of unovulated follicles and early regression of corpora lutea. In addition, the rate of fertilisation failure has been reduced by using an inducer of ovulation (i.e. GnRH) associated with intrauterine insemination. In summary, the application of recently acquired knowledge has resulted in relevant improvements in MOET programmes in small ruminants. PMID- 20003850 TI - Systems biology of embryogenesis. AB - The development of a complete organism from a single cell involves extraordinarily complex orchestration of biological processes that vary intricately across space and time. Systems biology seeks to describe how all elements of a biological system interact in order to understand, model and ultimately predict aspects of emergent biological processes. Embryogenesis represents an extraordinary opportunity (and challenge) for the application of systems biology. Systems approaches have already been used successfully to study various aspects of development, from complex intracellular networks to four dimensional models of organogenesis. Going forward, great advancements and discoveries can be expected from systems approaches applied to embryogenesis and developmental biology. PMID- 20003853 TI - Synthetic media for culture, freezing and vitrification of bovine embryos. AB - Media designed for the recovery, holding and cryopreservation of bovine and equine embryos are available from several commercial sources. In years past, some of these media contained bovine serum, although inclusion of serum in embryo transfer media is now largely discontinued due to issues relating to storage and biosecurity. Currently, bovine serum albumin (BSA) is included in most commercially manufactured media intended for use in embryo transfer (ET). Although BSA poses less risk than serum for the transmission of infectious disease, its inclusion still entails risk of viral contamination. The present review briefly describes the various components of ET media and the development of efficacious ET media containing no products of animal origin. An evaluation of the efficacy of recovery, holding, slow controlled freezing and vitrification media in both research and commercial ET embryo transfer settings is also presented. PMID- 20003854 TI - Use of equine chorionic gonadotrophin in synchronised AI of seasonal-breeding, pasture-based, anoestrous dairy cattle. AB - In seasonally calving dairy areas, a critical index of efficiency and profitability is the ability to maintain a short annual calving spread. In essence, this means that all cows need to conceive within 90 days of calving. Anovulatory anoestrus (AA) is a major barrier to reproductive performance under these conditions and the successful treatment of AA is an important aspect of reproductive intervention. The present paper reviews regimens used to treat AA in New Zealand and reports on a large-scale field trial investigating the effects of equine chorionic gonadotrophin (eCG) within a proven efficacious treatment protocol. The administration of eCG at the time of removal of a progesterone releasing device in an 8-day, oestradiol benzoate (EB)-based treatment protocol increased the percentage of cows in calf after 7 and 28 days of breeding. The interaction between cow age and treatment with eCG was significant, with cows over 5 years of age having higher conception rates (P = 0.002) and 7 day in-calf rates (P = 0.002) than cows on a standard 8-day treatment protocol without eCG. The addition of eCG to a standard 8-day EB/progesterone protocol can improve reproductive success in seasonally calving AA dairy cows. PMID- 20003855 TI - Use of knowledge regarding LH receptors to improve superstimulatory treatments in cattle. AB - Embryo transfer is a biotechnology that has been used worldwide to increase the production of offspring from female bovines. Treatments to induce multiple ovulations (superovulation) have evolved from superstimulatory protocols that depended upon detection of oestrus to treatments that synchronise follicle growth and ovulation, allowing for improved donor management and fixed-timed AI (FTAI). The protocols associated with FTAI facilitate animal handling and produce at least as many viably embryos as conventional treatment protocols that required detection of oestrus. Recent knowledge regarding LH receptors (LHR) and follicular development can be applied to improve embryo transfer protocols. In fact, improvements in the superstimulatory treatment called the 'P-36 protocol', which include hormones that stimulate LHR, indicate that adjustments related to LHR availability may increase bovine embryo yield compared with conventional protocols based on the detection of oestrus. PMID- 20003856 TI - Brief introduction to whole-genome selection in cattle using single nucleotide polymorphisms. AB - Genomic selection using single nucleotide polymorphisms (SNPs) is a powerful new tool for genetic selection. In cattle, SNP profiles for individual animals are generated using a small plastic chip that is diagnostic for up to 50 000 SNPs spaced throughout the genome. Phenotypes, usually averaged over offspring of bulls, are matched with SNP profiles of bulls mathematically so that animals can be ranked for siring desirable phenotypes via their SNP profiles. For many traits in dairy cattle, the rate of genetic improvement can be nearly doubled when SNP information is used in addition to current methods of genetic evaluation. Separate SNP analyses need to be developed for different populations (e.g. the system for Holsteins is not useful for Jerseys). In addition, the value of these systems is very dependent on the number of accurate phenotypes matched with SNP profiles; for example, increasing the number of North American Holstein bulls evaluated from 1151 to 3576 quadrupled the additional genetic gain in net merit from this approach. Thus, the available information will be insufficient to exploit this technology fully for most populations. However, once a valid SNP evaluation system is developed, any animal in that population, including embryos, can be evaluated with similar accuracy. Biopsying embryos and screening them via SNP analysis will greatly enhance the value of this technology by minimising generation intervals. PMID- 20003857 TI - Practical applications of new research information in the practice of bovine embryo transfer. AB - For more than 40 years, practitioners have sought to improve all aspects of commercial bovine embryo transfer. The development of new technologies for this industry has been substantial, with recent focus on cryopreservation techniques and the in vitro production of embryos fertilised with sexed spermatozoa. When these and other new technologies are developed, the following questions remain: (1) is said technology regulated or does it require licensing; and (2) is it applicable and, if so, is it financially feasible? Computer access to published research and the advancement of data software programs conducive to the industry for data procurement have been essential for helping practitioners answer these questions by enhancing their ability to analyse and apply data. The focus of the present paper is to aid commercial embryo transfer practitioners in determining new technologies that are available and whether they can be implemented effectively, benefiting their programs. PMID- 20003858 TI - Factors affecting fertilisation and early embryo quality in single- and superovulated dairy cattle. AB - Data on fertilisation and embryo quality in dairy cattle are presented and the main factors responsible for the low fertility of single-ovulating lactating cows and embryo yield in superovulated dairy cattle are highlighted. During the past 50 years, the fertility in high-producing lactating dairy cattle has decreased as milk production increased. Recent data show conception rates to first service to be approximately 32% in lactating cows, whereas in heifers it has remained above 50%. Fertilisation does not seem to be the principal factor responsible for the low fertility in single-ovulating cows, because it has remained above 80%. Conversely, early embryonic development is impaired in high-producing dairy cows, as observed by most embryonic losses occurring during the first week after fertilisation. However, in superovulated dairy cattle, although fertilisation failure is more pronounced, averaging approximately 45%, the percentage of fertilised embryos viable at 1 week is quite high (>70%). Among the multifactorial causes of low fertility in lactating dairy cows, high feed intake associated with low concentrations of circulating steroids may contribute substantially to reduced embryo quality. Fertilisation failure in superovulated cattle may be a consequence of inappropriate gamete transport due to hormonal imbalances. PMID- 20003859 TI - [Preventive resection of incidentally perioperatively found Meckel diverticulum]. PMID- 20003860 TI - [Perforation of Meckel's diverticulum with intraabdominal hemorrhage and periappendicular inflammation]. AB - Perforation of Meckel's diverticulum (MD) is a rare and serious complication. The authors report a case of a 12-year-old girl who presented with acute abdominal pain and anaemia. Haemoperitoneum, perforated MD and peri-appendicular inflammation were found during laparotomy. The patient was treated with resection of a segment of ileum bearing the diverticulum and appendectomy. Postoperative recovery was uneventful. Pathological examination showed a perforated peptic ulcer with acute peritonitis and periappendicitis. PMID- 20003861 TI - [A bleeding gastrointestinal stromal tumour in a Meckel's diverticulum]. AB - We present at rare case of severe upper gastrointestinal bleeding in a 57-year old patient due to a gastrointestinal stromal tumour located in a Meckel's diverticulum (MD) without ectopic tissue. We discuss indications and pro et cons of prophylactic resection of MD. PMID- 20003862 TI - [Characterization of complex renal cysts]. AB - Complex renal cysts represent a clinical problem, since it is often difficult to exclude malignancy. The Bosniak classification uses computed tomography (CT) to categorize cystic renal lesions. Category I and II are presumed to be benign, and III and IV potentially malignant. Category IIF cysts are lesions that may develop into malignancy. Studies exploring whether the IIF category solves the clinically important problem of differentiating between benign and malignant cysts are needed. PMID- 20003863 TI - [Vitamin D, cardiovascular disease and mortality in patients with kidney disease]. AB - Chronic kidney disease is associated with an increased risk of cardiovascular disease. Vitamin D deficiency is common in patients with chronic kidney disease. In epidemiological studies, vitamin D deficiency and absence of treatment with vitamin D is associated with increased cardiovascular mortality. Several possible mechanisms may explain how vitamin D can influence the development of cardiovascular disease. Clinical intervention studies are needed to clarify whether treatment with vitamin D decreases the risk of cardiovascular disease in chronic kidney disease. PMID- 20003864 TI - [Adverse events causing discontinuation of donepezil for Alzheimer's dementia]. AB - INTRODUCTION: The objective was to investigate the frequency of adverse events (AE) as a cause for discontinuation of donepezil treatment for Alzheimer's dementia (DAT) in a geriatric memory unit. MATERIAL AND METHODS: Five-year retrospective study of 123 donepezil-treated patients diagnosed with DAT or mixed dementia in a geriatric memory unit. The material covers all patients treated with donepezil and surveyed for 12 months in the memory unit during the period from 14th March 2001 to 7th April 2006. RESULTS: Among the 123 patients, 106 (86%) suffered from DAT and 17 (14%) suffered from mixed dementia. A total of 100 (81%) were female while 23 (19%) were male. The median age was 84 years. In all, 26 (21%) patients discontinued treatment due to AE. The most frequent AE were nausea/vomiting, diarrhoea and loss of appetite. CONCLUSION: In 21% of the cases treatment was discontinued within 12 months due to AE. The most frequent AEs were nausea/vomiting, diarrhoea and loss of appetite. For the most part discontinuation took place within the initial three months. However, some treatments were not discontinued until after six months; consequently, treatment should be supervised and patients should have easy access to a memory unit in the case of AE. PMID- 20003865 TI - [Schistosomiasis in Danish travellers and immigrants]. AB - INTRODUCTION: The purpose of the study was to describe symptoms and diagnosis of schistosomiasis in travellers and immigrants based on patients diagnosed and treated at the State University Hospital in Copenhagen in 2003-2008. MATERIAL AND METHODS: Retrospective review of patient records. RESULTS: A total of 49 patients (39 travellers and ten immigrants) were diagnosed with schistosomiasis. All patients except one were infected in Africa. There was considerable variation in clinical manifestations between travellers and immigrants. Eggs were detected in urine, faeces and/or biopsy from 18 patients, whereas 31 patients were diagnosed solely by serology. DISCUSSION: Travellers with schistosomiasis are often asymptomatic or have unspecific symptoms, while immigrants may present with severe complications to the infection. Because symptoms can develop years after the infection and due to the risk of severe complications, even asymptomatic patients should be treated. Treatment is simple and has few side effects. PMID- 20003866 TI - [A survey of research funding at Danish regional hospitals]. AB - INTRODUCTION: According to recently passed Danish legislation, all Danish hospitals are obliged to take part in scientific research. As data on financial support for research activities are lacking, we assessed the resources allocated to research from the budget of the central hospital management as a percentage of the total budget at Danish regional hospitals in 2007. MATERIAL AND METHODS: A postal survey was conducted at 13 hospitals in the Western part of Denmark. The questionnaire comprised items in the following major categories: 1) budget allocated specifically for research and travel grants; 2) employment of scientific and technical support staff; 3) facilities and equipment for research; and 4) research dissemination. RESULTS: Questionnaires were returned from 11 hospitals. Six hospitals reported to have dedicated fixed amounts on the budget for research, exact figures were reported in four cases only equivalent to 0.1%, 0.3%, 0.3% and 0.6% of the total budget. Most hospitals had associate professors, but only five had full professors. Seven hospitals supplied laboratories and technical facilities, eight hospitals held staff-meetings on a regular basis and four published an annual report on research activities. CONCLUSION: In the majority of regional hospitals in Western Denmark, less than 0.3% of the total budget administered by the central hospital management was allocated specifically for research. These figures, however, may not be accurate as individual departments may allocate additional resources from local budgets. We recommend that regional hospitals define research strategies and allocate the necessary funding in their budgets. PMID- 20003867 TI - [Keystone flap for reconstruction of skin defects]. AB - INTRODUCTION: The keystone design perforator island flap was first described in 2003. It is a local flap based on axial perforators from the underlying structures. It is designed as a curvilinear shaped trapezoidal design flap which is essentially two V-Y flaps end to end. Most reports are of the lower limbs, but it has been used successfully almost all over the body. MATERIAL AND METHODS: We report our experience in 21 patients, who underwent excision of skin malignancies and reconstruction of the defect with a keystone flap (24 flaps in total). RESULTS: Only few complications were seen. Two patients had signs of infection at the border of the flap and were treated with local and/or oral antibiotics. In two cases a small necrosis developed at the border of the flap. In both cases, it healed conservatively. CONCLUSION: The keystone flap has proven to be very reliable with only few reported flap losses. Furthermore, it is technically easy to perform and thus may be performed by trainee surgeons as well as senior surgeons. The cosmetic result is excellent and far superior to the alternative skin grafting with its color differences and contour defect. In addition, many patients may be treated as day-only surgery or discharged from the hospital after only one day. PMID- 20003868 TI - [Ranolazine--new treatment of chronic stable angina pectoris]. AB - Ranolazine sustained-release tablets were recently approved in the EU for chronic stable angina as add-on therapy when symptoms are not controlled with first-line agents. The mechanism of action is thought to involve inhibition of late sodium influx in the heart, which can reduce abnormalities of contractility and repolarisation associated with ischaemia. Ranolazine increases the exercise capacity, reduces angina, and diminishes the use of nitroglycerine. The drug has an excellent safety profile and may be a valuable addition to the treatment of chronic stable angina. PMID- 20003869 TI - [The influence of allergic rhinitis on asthma]. AB - Allergic rhinitis presents as intermittent or persistent nasal symptoms with sneeze, blockage and secretions. Rhinitis affects more that 20% of the population and is more frequent in younger persons. Allergic rhinitis influences all aspects of quality of life, school performance and work. Most asthmatics suffer from rhinitis and many rhinitis patients suffer from asthma. Patients with rhinitis should be evaluated for asthma and patients with asthma should be evaluated for rhinitis. A combined strategy should be applied in the treatment of upper and lower airways. PMID- 20003872 TI - Modified biplanar chevron osteotomy. AB - BACKGROUND: The surgical treatment of hallux valgus with an increased distal metatarsal articular angle (DMAA) should include the correction of this angle to maintain a congruent joint. The purpose of this study was to report our results with this procedure. MATERIALS AND METHODS: From January 2000 until December 2006, 23 feet (13 patients) with an increased DMAA were operated upon by the same surgeon using a biplanar chevron osteotomy. The technique was modified making the plantar cut more horizontal and only removing a wedge from the dorsal cut. Clinical and radiographic evaluation was made after a mean followup of 56 months. The AOFAS score for the hallux was used and angular deformities were measured. The patients had a mean age of 51 years and were all female. RESULTS: A good functional result was achieved with a mean AOFAS score of 87. An improvement of the angular deformities was noted in all patients. The mean hallux valgus angle improved from 28 degrees pre-op to 14 degrees post-op, the intermetatarsal angle from 12 degrees to 7 degrees and the DMAA from 20 degrees to 7 degrees. All patients except one were satisfied with the outcome of the procedure. Minimally symptomatic AVN of the metatarsal head was observed in one foot. CONCLUSIONS: We conclude that this procedure is a good choice for the treatment of symptomatic hallux valgus with an increased DMAA, producing a good clinical and radiographic result. PMID- 20003873 TI - Comparison of the proximal chevron and Ludloff osteotomies for the correction of hallux valgus. AB - BACKGROUND: Although several studies have described good results of proximal chevron and Ludloff osteotomies, there have been no studies comparing the results of these two techniques at a single institution. MATERIALS AND METHODS: We consecutively evaluated 46 patients who underwent proximal chevron osteotomies and 52 patients who underwent Ludloff osteotomies. Patients were evaluated by preoperative and postoperative weight bearing radiographs and the American Orthopaedic Foot and Ankle Society (AOFAS) hallux MP score. RESULTS: Both groups had similarly high AOFAS scores and good correction by radiographic parameters. No statistically significant differences were found with respect to correction of hallux valgus angle (HVA) and intermetatarsal angle (IMA) between the two groups. Significant shortening of the first metatarsal was found after Ludloff osteotomy (p < 0.05). At 6 weeks after surgery, the pain subscore was significantly lower in the proximal chevron group than in the Ludloff group (p < 0.05). CONCLUSIONS: The proximal chevron and Ludloff osteotomies yielded equivalent clinical and radiological results. The Ludloff osteotomy with lag screw fixation is more stable and does not require postoperative hardware removal, although it is technically demanding and has a tendency toward greater shortening of the first metatarsal. PMID- 20003874 TI - Minimally invasive treatment of distal tibial fractures with locking and non locking plates. AB - BACKGROUND: The purpose of this study was to evaluate the clinical performances of stainless steel nonlocking plates and titanium locking plates in minimally invasive medial plating of extra-articular metaphyseal fractures of the distal tibia. MATERIALS AND METHODS: Between 2004 and 2006, 43 patients who sustained closed fractures of the distal tibia metaphysis were managed with either a stainless steel nonlocking plate (Group 1, n = 21) or a titanium locking plate (Group 2, n = 22). Clinical and radiographic data were retrospectively reviewed. Function was assessed with use of the American Orthopaedic Foot and Ankle Society ankle-hindfoot instrument. The average followup period was 25 months. RESULTS: The average AOFAS foot and ankle scores in Group 1 and Group 2 at final followup were 85 and 81, respectively. Fracture reduction was anatomical or nearly anatomical without angular displacement in all cases except one patient in Group 2. (p > 0.05) The average time to full, unprotected weightbearing in Group 1 and Group 2 were 15 weeks and 18 weeks, respectively (p < 0.05). CONCLUSION: Minimally invasive medial plating with titanium locking plates resulted in prolonged secondary healing both in comminuted and simple fracture patterns compared to conventional stainless steel nonlocking plates. We believe that in biological fixation of distal tibial fractures, similarly good results may be obtained with both materials as the locking technology had a greater effect on stability. PMID- 20003875 TI - New radiographic parameters assessing forefoot abduction in the adult acquired flatfoot deformity. AB - BACKGROUND: Stage II flatfoot secondary to posterior tibial tendon insufficiency may be subclassified into mild (IIa) and severe (IIb) deformity based on the degree of talonavicular abduction. Current assessment of this abduction is difficult. We hypothesized that two new anteroposterior radiographic parameters, the lateral talonavicular incongruency angle (IA) and incongruency distance (ID) would demonstrate good reliability, correlate with current abduction parameters, and differ in IIb deformity, IIa deformity, and controls. MATERIALS AND METHODS: Preoperative radiographs for consecutive patients undergoing flatfoot reconstruction were reviewed and subdivided into those with a Stage IIb (n = 32) or Stage IIa (n = 8) deformity. A third group of patients without flatfoot served as control (n = 30). Radiographs were measured blindly by two investigators. Reliability was assessed with intraclass correlation coefficients (ICC), correlation with existing parameters with Pearson coefficients, and comparison between groups with analysis of variance. RESULTS: The mean intrarater and interrater ICC's for the IA (0.88 and 0.81, respectively) were high. The IA correlated well with the coverage angle (r = 0.86) and uncoverage percent (r = 0.76). The IA was higher in the IIb versus IIa patients (p = 0.007) and in the IIb group versus control (p < 0.001). The ID demonstrated excellent reliability (ICC's of 0.83 and 0.83), but correlated poorly with the two other abduction parameters (r = -0.59 and -0.49) and failed to differentiate between the three groups (p = 0.0528). CONCLUSION: This data suggests that the IA is reliable and may help subclassify Stage II flatfoot deformity. PMID- 20003876 TI - Risk factors for failure of transmetatarsal amputation in diabetic patients: a cohort study. AB - INTRODUCTION: This study analysed the factors affecting wound healing of a transmetatarsal amputation (TMA) in patients with diabetes. MATERIALS AND METHODS: Twenty-one patients who failed a TMA and were revised to transtibial amputation (TTA) within the first year were compared with a matched cohort of 21 successful TMA's. The factors compared included demographics, systemic issues, and local conditions in the foot. Chi-square comparisons were used for group data, and Analysis of Variance (ANOVA) for numeric data. RESULTS: Blood glucose control as measured by HbA1c was the most important single factor predicting the success of TMA. Need for debridement after TMA was also found to be a significant predictor of failure of TMA. There was a trend towards duration of ulcer prior to TMA and smoking being significant. All other variables, including vascular status or renal failure were not significantly different between the two groups. CONCLUSION: The primary factor determining the success of a TMA was the quality of glucose control. The results of this study can be extrapolated to diabetic patients undergoing other types of surgery, with preoperative diabetic control as measured by HbA1c being an important determinant of the outcome of surgery. As a result of this study we currently do not perform any elective, trauma or emergency surgery on diabetic patients with an HbA1c of over 8 unless the need for surgery is to save life or limb. PMID- 20003877 TI - Intraoperative mechanical bone strength determination in tibiotalocalcaneal fusion: a biomechanical investigation. AB - BACKGROUND: Bone strength is currently measured with indirect techniques. We investigated the use of an intraoperative mechanical measurement for local bone strength determination and prediction of intramedullary-nail fusion failure. We investigated whether intraoperative local bone strength determination may be useful to the surgeon in predicting intramedullary nail hindfoot fusion performance. MATERIALS AND METHODS: In seven human specimens, bone mineral density (BMD) was determined with qCT. A device (DensiProbe) specially devised for nailed tibiotalocalcaneal arthrodesis (TTCA) was inserted at the intended calcaneal screw sites of an intramedullary nail, and the cancellous break-away torque was measured. The constructs were then cyclically loaded to failure in dorsiflexion-plantarfexion. RESULTS: The BMD range was wide (42.8 to 185.9 mg HA/cm(3)). The proximal-screw site peak torque was 0.47 to 1.61 Nm; distal-screw site peak torque was 0.24 to 1.06 Nm. The number of cycles to failure correlated with peak torque both proximally (p = 0.021; r(2) = 0.69) and distally (p = 0.001; r(2) = 0.92). Proximally, peak torque did not correlate with BMD (p = 0.060; r(2) = 0.54); distally, it correlated significantly (p = 0.003; r(2) = 0.86). CONCLUSION: DensiProbe measurements can be used in the hindfoot to assess bone strength. In this study, specimens that failed early could be identified. However, in clinical practice fusion failure is multifactorial in origin, and failure prediction cannot be based upon peak torque measurements alone. CLINICAL RELEVANCE: The technique described here may be of use to give an intraoperative decision aid to predict intramedullary nail hindfoot fusion performance. PMID- 20003878 TI - Kinematic analysis of runners with achilles mid-portion tendinopathy. AB - BACKGROUND: Despite anecdotal evidence linking overpronation to the onset of Achilles tendinopathy (AT), there is little conclusive evidence of a particular movement pattern of the lower extremity associated with this injury. Therefore, the objective of the present study was to observe differences in the kinematic profiles of healthy runners (CON) and runners with mid-portion Achilles tendinopathy (ATG). MATERIALS AND METHODS: In this cross-sectional analysis, 48 male height and weight matched subjects were invited to participate: 27 with mid portion Achilles tendon pain and 21 asymptomatic controls. Subjects underwent lower extremity clinical examination, then ran barefoot for 10-trials at a self selected pace. A 3D motion capture system analysed tri-plane kinematic data for the lower extremity. RESULTS: The ATG displayed significantly greater sub-talar joint eversion displacement during mid-stance of the running gait (13 +/- 3 degrees vs. 11 +/- 3 degrees; p = 0.04). Trends were observed such that the ATG showed lower peak dorsiflexion velocity (300 +/- 39 degrees/s vs. 330 +/- 59 degrees/s; p = 0.08) and greater overall frontal plane ankle joint range of motion (45 degrees +/- 7 vs. 41 degrees +/- 7; p = 0.09). CONCLUSION: We found an increase in eversion displacement of the sub-talar joint in runners with Achilles mid-portion tendinopathy. Based on the findings from this study, there is evidence that devices used to control sub-talar eversion may be warranted in patients with Achilles mid-portion tendinopathy who demonstrate over-pronation during mid-stance of the running gait. PMID- 20003879 TI - Sciatic nerve blockade: a survey of orthopaedic foot and ankle specialists in North America and the United Kingdom. AB - BACKGROUND: Sciatic nerve blocks are used to reduce post-operative pain and allow early discharge for patients undergoing foot and ankle surgery. This study aimed to identify the utilization of this procedure in the US and UK and to establish the standard of care with respect to the level of anesthesia that the patient is under and use of ultrasound localization when performing sciatic nerve blocks. MATERIALS AND METHODS: A survey of current committee members of AOFAS and members of BOFAS. RESULTS: Two hundred sixty-three surgeons were contacted with a response rate of 44%. Eighty-two percent commonly used a sciatic nerve blockade. Sixty-nine percent never or only sometimes used ultrasonography and variable levels of nerve stimulation were used. Forty-two percent where happy to have the block performed under full anesthesia. There were significant differences between British and American practices regarding the level of nerve stimulation and the level of anesthesia used. The most common complication cited was prolonged anesthesia of which the vast majority spontaneously resolved. Performing blocks awake or sedated did not seem to alter number of complications seen. CONCLUSION: This study represents a current practice review of sciatic nerve blocks performed amongst senior foot and ankle surgeons. Although no absolute consensus has been reached as to the use of ultrasound or whether the patient needs to be awake for the procedure, it is clear that the standard of care does not mandate either of these. The differences between US and UK practice are probably cultural and do not appear to affect the number of complications encountered. PMID- 20003880 TI - Surgical anatomy of the proximal release of the gastrocnemius: a cadaveric study. AB - BACKGROUND: The importance of isolated gastrocnemius contracture in disorders of the foot and ankle has been established in recent years. The aim of this study was to describe the proximal anatomical approach to the medial and lateral heads of gastrocnemius and to compare the sizes of the medial and lateral heads of the gastrocnemius. MATERIALS AND METHODS: 15 cadaveric extremities were dissected using a posterior approach 1 cm below the level of the skin crease. Proximity of cutaneous nerves and major vessels was noted. The heads of the gastrocnemius were dissected from their origin and the cross sectional anatomy was assessed. RESULTS: Approach to the medial head of gastrocnemius was safe. Conversely the variable anatomy of the nerves in the approach to the lateral head meant that extreme care must be taken if complications are to be avoided. The aponeurosis of the medial head of gastrocnemius was 2.4 times the cross-sectional area compared to the lateral head. CONCLUSION: In this study we describe a safe posterior approach to the medial aponeurosis of the gastrocnemius and also describe the different sizes of the medial and lateral gastrocnemius heads. CLINICAL RELEVANCE: We conclude that the approach to the medial head of gastrocnemius is free from neurovascular structures and that release of the medial head alone may be efficacious in the operative treatment of isolated gastrocnemius tightness that has failed non-operative treatment. PMID- 20003881 TI - Tendon transfer fixation in the foot and ankle: a biomechanical study. AB - BACKGROUND: Tendon transfers are often used in foot and ankle surgery. Different fixation devices and techniques have been described. The most recently developed ones are bone anchors and interference screws. MATERIALS AND METHODS: A biomechanical study was designed to compare tendon transfer fixation, using Corkscrew bone anchors 5 x 15.5 mm and Bio-interference screws (8 x 23 mm). Fifteen fresh cadaver specimens underwent both fixation techniques for split anterior tibial tendon transfer at the cuboid bone. All the specimens underwent standardized X-rays in order to evaluate mineral bone density using a standardized measurement system. All were tested until maximal load to failure. RESULTS: The ultimate load to failure of the tendon secured to the cuboid using anchors was 103 N (SD, 52), compared with 150 N (SD, 68) for tendons secured to the bone with interference screws (p = 0.003). No difference was found between the techniques that could be related to bone density. CONCLUSION: Interference screws provided greater strength than bone anchors. CLINICAL RELEVANCE: This study demonstrated increased strength in securing bone to tendon in vitro for SPLATT tendon transfer with interference screws as compared to suture anchors. PMID- 20003882 TI - Comparison of calcaneal fixation of a retrograde intramedullary nail with a fixed angle spiral blade versus a fixed-angle screw. AB - BACKGROUND: Retrograde intramedullary nailing is an established technique for tibiotalocalcaneal arthrodesis (TTCA). In poor bone stock (osteoporosis, neuroarthropathy), device fixation in the hindfoot remains a problem. Fixed-angle spiral-blade fixation of the nail in the calcaneus could be useful. MATERIALS AND METHODS: In seven matched pairs of human below-knee specimens, bone mineral density (BMD) was determined, and TTCA was performed with an intramedullary nail (Synthes Hindfoot Arthrodesis Nail HAN Expert Nailing System), using a conventional screw plus a fixed-angle spiral blade versus a conventional screw plus a fixed-angle screw, in the calcaneus. The constructs were subjected to quasi-static loading (dorsiflexion/plantarflexion, varus/valgus, rotation) and to cyclic loading to failure. Parameters studied were construct neutral zone (NZ) and range of motion (ROM), and number of cycles to failure. RESULTS: With dorsiflexion/plantarflexion loading, the screw-plus-spiral-blade constructs had a significantly smaller ROM in the quasi-static test (p = 0.028) and early in the cyclic test (p = 0.02); differences in the other parameters were not significant. There was a significant correlation between BMD and cycles to failure for the two screw constructs (r = 0.94; p = 0.002) and for the screw-plus-spiral-blade constructs (r = 0.86; p = 0.014). CONCLUSION: In TTCA with a HAN Expert Nailing System, the use of a calcaneal spiral blade can further reduce motion within the construct. This may be especially useful in poor bone stock. CLINICAL RELEVANCE: Results obtained in this study could be used to guide the operating surgeon's TTCA strategy. PMID- 20003883 TI - Percutaneous bone biopsy to distinguish osteomyelitis from charcot osteoarthropathy: two case reports. PMID- 20003884 TI - Tibial shaft fracture with concomitant syndesmotic injury: a case report. PMID- 20003885 TI - Replantation of severed foot at the chopart joint: a case report. PMID- 20003886 TI - Periprosthetic fracture after total ankle replacement: surgical technique. PMID- 20003887 TI - Technical tip: fixation of the scarf metatarsal osteotomy with a proximal buttress and a standard distal screw. PMID- 20003888 TI - The problem is background, not signal. PMID- 20003889 TI - Translational modulation of proteins expressed from bicistronic vectors. AB - Bicistronic vectors are useful tools for exogenous expression of two gene products from a single promoter element; however, reduced expression of protein from the second cistron compared with the first cistron is a common limitation to this approach. To overcome this limitation, we explored use of dihydrofolate reductase (DHFR) complementary DNA encoded in bicistronic vectors to induce a second protein of interest by methotrexate (MTX) treatment. Previous studies have demonstrated that levels of DHFR protein and DHFR fusion protein can be induced translationally following MTX treatment of cells. We demonstrated that in response to MTX treatment, DHFR partner protein in a bicistronic construct is induced for longer periods of time when compared with endogenous DHFR and DHFR fusion protein, in vitro and in vivo. Using rapamycin pretreatment followed by MTX treatment, we also devised a strategy to modulate levels of two proteins expressed from a bicistronic construct in a cap-independent manner. To our knowledge, this is the first report demonstrating that levels of proteins in DHFR based bicistronic constructs can be induced and modulated using MTX and rapamycin treatment. PMID- 20003890 TI - Fluorescence imaging of fast retrograde axonal transport in living animals. AB - Our purpose was to enable an in vivo imaging technology that can assess the anatomy and function of peripheral nerve tissue (neurography). To do this, we designed and tested a fluorescently labeled molecular probe based on the nontoxic C fragment of tetanus toxin (TTc). TTc was purified, labeled, and subjected to immunoassays and cell uptake assays. The compound was then injected into C57BL/6 mice (N = 60) for in vivo imaging and histologic studies. Image analysis and immunohistochemistry were performed. We found that TTc could be labeled with fluorescent moieties without loss of immunoreactivity or biologic potency in cell uptake assays. In vivo fluorescent imaging experiments demonstrated uptake and retrograde transport of the compound along the course of the sciatic nerve and in the spinal cord. Ex vivo imaging and immunohistochemical studies confirmed the presence of TTc in the sciatic nerve and spinal cord, whereas control animals injected with human serum albumin did not exhibit these features. We have demonstrated neurography with a fluorescently labeled molecular imaging contrast agent based on the TTc. PMID- 20003891 TI - Extra domain B fibronectin as a target for near-infrared fluorescence imaging of rheumatoid arthritis affected joints in vivo. AB - Abstract We investigated a molecular imaging approach for the detection of collagen-induced arthritis in rats by targeting the extra domain B (ED-B) of the extracellular matrix protein fibronectin. ED-B is a highly conserved domain (identical in human and rats) that is produced by alternative splicing during embryonic development and during vascular remodeling such as angiogenesis. The hallmark of rheumatoid arthritis is synovitis leading to both angiogenesis in the synovium and the promotion of cartilage and bone disruption. For in vivo diagnostics, the ED-B-binding single-chain antibody fragment AP39 was used as a targeting probe. It was covalently linked to the near-infrared dye tetrasulfocyanine (TSC) to be visualized by near-infrared fluorescence imaging. The resulting AP39-TSC conjugate was intravenously administered to rats with collagen-induced arthritis and the respective controls. Ovalbumin-TSC was used as control conjugate. Optical imaging over a time period of 24 hours using a planar imaging setup resulted in a clear enhancement of fluorescence intensity in joints with moderate to severe arthritis compared with control joints between 3 and 8 hours postinjection. Given that AP39 is a fully human antibody fragment, this molecular imaging approach for arthritis detection might be translated to humans. PMID- 20003892 TI - Toxicity of organic fluorophores used in molecular imaging: literature review. AB - Fluorophores are potentially useful for in vivo cancer diagnosis. Using relatively inexpensive and portable equipment, optical imaging with fluorophores permits real-time detection of cancer. However, fluorophores can be toxic and must be investigated before they can be administered safely to patients. A review of published literature on the toxicity of 19 widely used fluorophores was conducted by searching 26 comprehensive biomedical and chemical literature databases and analyzing the retrieved material. These fluorophores included Alexa Fluor 488 and 514, BODIPY FL, BODIPY R6G, Cy 5.5, Cy 7, cypate, fluorescein, indocyanine green, Oregon green, 8-phenyl BODIPY, rhodamine 110, rhodamine 6G, rhodamine X, rhodol, TAMRA, Texas red, and Tokyo green. Information regarding cytotoxicity, tissue toxicity, in vivo toxicity, and mutagenicity was included. Considerable toxicity-related information was available for the Food and Drug Administration (FDA)-approved compounds indocyanine green and fluorescein, but published information on many of the non-FDA-approved fluorophores was limited. The information located was encouraging because the amounts of fluorophore used in molecular imaging probes are typically much lower than the toxic doses described in the literature. Ultimately, the most effective and appropriate probes for use in patients will be determined by their fluorescent characteristics and the safety of the conjugates. PMID- 20003894 TI - Health Protection Agency publishes Annual Evidence Update focussing on HIV in children and adolescents. PMID- 20003895 TI - The effectiveness of behavioural and psychosocial HIV/STI prevention interventions for MSM in Europe: A systematic review. AB - Given the need of programme planners and policy makers for descriptions of specific interventions and quantitative estimates of intervention effects to make informed decisions concerning prevention funding and research, there is a need for a systematic review that updates the current knowledge base about HIV/STI preventive interventions targeted at men who have sex with men (MSM) in Europe. The aim was to summarise and assess the effectiveness of HIV/STI prevention interventions for MSM living in Europe, and to identify intervention characteristics associated with effectiveness as well as potential gaps in the evidence base. A systematic search for relevant literature in eight international databases and in reference lists of relevant reviews and included studies was performed. Studies were selected according to pre-specified criteria and appraised for risk of bias. We summarised results using tables and calculated effect estimates for sexual behaviour outcomes. Results from six controlled studies, involving a total of 4,111 participants at entry from four different European countries were summarised. The results showed that there was 'high' or 'unclear' risk of bias in one or more of the assessed domains in all studies. The pooled effect estimate of the four interventions for which data were available suggested that MSM who participate in HIV/STI prevention initiatives may be somewhat less likely to report unprotected anal intercourse (UAI). The evidence base was insufficient to examine characteristics of interventions most closely associated with magnitude of effect and to draw solid conclusions about unique gaps in the evaluation literature. Despite the maturity of the HIV epidemic, rigorous outcome evaluations of any form of behavioural HIV/STI intervention for MSM in Europe are scarce. The results point to possible short term effects of interventions in terms of reductions in the proportion of MSM who engage in UAI, but the paucity of controlled studies demonstrates the need for research in this area. There is an overall deficit in outcome evaluations of interventions aimed at reducing HIV/STI risk behaviour among MSM in Europe. Designing behavioural HIV/STI preventive strategies to avert new infections, and the evaluation of such prevention programmes for MSM is an important component of a comprehensive HIV/STI containment strategy across the continuum of prevention and care. PMID- 20003896 TI - HIV risk behaviour knowledge, substance use and unprotected sex in men who have sex with men in Tallinn, Estonia. AB - This study examines HIV risk behaviour knowledge, substance use and unprotected sex in a sample of 79 men who have sex with men (MSM) in Tallinn, Estonia. Median age of the study population was 30 years (range 18-62 years); 35 were bisexual; 56 answered correctly to at least 10 out of 13 questions about HIV risk behaviours; 23 consumed more than seven alcoholic drinks in the week before the survey; nearly half (n=34) of the participants reported some illicit drug use in the past 12 months; 40 did not use a condom regularly in the 12 months preceding the survey, and 41 did not use a condom during their last sexual intercourse. Alcohol consumption in the week before the survey was negatively associated with condom use during last intercourse (RR 0.48; 95% CI 0.41-0.56). Use of illicit drugs varied significantly by ethnicity (p-value = 0.02). Multivariable analysis showed that higher consumption of alcohol in the week before the survey could be predicted by education, age group and sexual orientation. In conclusion, socio demographic factors such as education, age, ethnicity and sexual orientation may affect HIV risk behaviour knowledge, sexual behavior and substance use among MSM in Estonia, and need to be taken into consideration for targeted HIV prevention. PMID- 20003897 TI - HIV bio-behavioural survey among men who have sex with men in Barcelona, Bratislava, Bucharest, Ljubljana, Prague and Verona, 2008-2009. AB - Data from 23 European countries show that the annual number of HIV diagnoses in men who have sex with men (MSM) increased by 86% between 2000 and 2006. This paper reports the main preliminary results of a bio-behavioural survey in MSM with a specific focus on HIV prevalence and use of United Nations General Assembly Special Session (UNGASS) indicators in six cities in Southern and Eastern Europe. Time-location sampling (TLS) was used. A total number of 2,356 questionnaires and 2,241 oral fluid samples were collected (invalid samples 4.1%). The data show different socio-demographic patterns across countries regarding age, level of education, living conditions, living area and self identity. Southern European cities had the highest percentage of people who had tested for HIV and collected the result. More than 50% of respondents in the sample from Barcelona reported having used a condom last time they had anal sex (57.2%), whilst in all other cities this proportion was below 50%. The cities with the highest HIV prevalence in MSM were Barcelona (17.0%) and Verona (11.8%) whilst lower percentages were reported in Bratislava (6.1%), Bucharest (4.6%), Ljubljana (5.1%) and Prague (2.6%). The low prevalence in Eastern European cities is encouraging. However, with the level of high-risk sexual behaviour documented and the lower frequency of HIV test seeking behaviour, there is a clear risk of an increase in HIV transmission. PMID- 20003898 TI - Lymphogranuloma venereum in Europe, 2003-2008. AB - Lymphogranuloma venereum, caused by the L serovars of Chlamydia trachomatis, emerged in Europe in 2003 and a series of outbreaks were reported in different countries. The infection presents as a severe proctitis in men who have sex with men, many of whom are co-infected with HIV and other sexually transmitted infections. This paper reviews the number of cases reported over a five year period, from 2003 to 2008, from countries that were part of the European Surveillance of Sexually Transmitted Infections (ESSTI) network. Reports were received from Belgium, Denmark, France, Germany, the Netherlands, Portugal, Spain, Sweden, and the United Kingdom. It appears that after five years the characteristics of the patients infected has overall remained unchanged, although the total number of cases has increased and more countries in Europe have now identified cases of LGV. PMID- 20003900 TI - Recently acquired HIV infection in men who have sex with men (MSM) in France, 2003-2008. AB - An increase in the number of new HIV diagnoses among men who have sex with men (MSM) has been observed in several countries in the early 2000s. In this article, we explore the trends in MSM in France between 2003 and 2008. To estimate the number of MSM newly diagnosed with HIV, we take into account the reporting delay, underreporting and missing data for HIV case notification. To identify recent infections (RI) (acquired an average of six months before diagnosis), we used an enzyme immunoassay for recent HIV-1 infections (EIA-RI) which has been performed routinely for new HIV diagnoses since 2003. Multivariate analysis was used to identify factors associated with RI. We estimate that between 1,900 and 2,400 MSM have been newly diagnosed with HIV every year: the proportion of MSM among all newly diagnosed with HIV cases has increased from 25.2% (95% confidence interval (CI): 23.3-27.1) in 2003 to 37.0% (95% CI: 35.2-38.7) in 2008 and was stable during the period 2006-2008. In 2008, the rate of newly diagnosed HIV cases per 10,000 MSM living in France was 72.5. The proportion of non-B subtypes of HIV-1 among cases diagnosed in MSM was 11.7% (2003-2008). The assessment of RI was performed for 4,819 MSM newly diagnosed with HIV in 2003-2008. Of these, 47.6% (95%CI = 46.2-49.0) (2,295 cases) were shown to have been recently infected. The risk of RI was greater for those of French nationality (adjusted odds ratio (aOR) =1.6 [95% CI: 1.4-1.9]), those with high economic status (aOR =1.4 [95% CI: 1.2 1.8]), those tested after a risk exposure (aOR =1.6[95% CI: 1.3-1.8]) or after presenting with clinical symptoms or abnormal biological markers (aOR =1.8 [95% CI: 1.5-2.0]), those who had tested for HIV three or more times during their life time (aOR =4.2 [95% CI: 3.4-5.2]) and those living in the Paris area (aOR =1.2 [95% CI: 1.0-1.3]). The risk of RI decreased with age. The HIV situation among MSM living in France is a cause of concern, despite the prevention campaigns dedicated to this highly educated sub-population. PMID- 20003901 TI - Symptomatic primary HIV infection in a 49-year-old man who has sex with men: beware of the window phase. AB - A 49-year-old man with a history of receptive unprotected anal intercourse with multiple anonymous men presented with a symptomatic primary HIV infection. Upon his initial visit the rapid HIV antibody screening test was negative but a p24 antigen test suggested a highly infectious phase in the HIV infection. An immunoblot assay confirmed the HIV diagnosis only 14 days later. Recent infections are characterised by a highly infectious phase and, if gone unnoticed, can have a large contribution to the ongoing transmission of HIV. Healthcare providers should be aware of primary HIV infection and the pitfalls in its diagnosis. PMID- 20003903 TI - Viral hepatitis, HIV, human herpes virus and Treponema pallidum infection in haemodialysis patients from Kosovo, 2005. AB - The serological status of hepatitis viruses and other infectious diseases in the 66 dialysed patients of one haemodialysis unit in Kosovo were studied, comparing the data with a large group of blood donors and out-patients. All dialysed patients were hepatitis A virus (HAV) positive. Prevalence of hepatitis B surface antigen (HBsAg), hepatitis B surface antibodies (anti-HBs), and hepatitis B core antibodies (anti-HBc) was 14 of 66, 21% (95% confidence interval (CI): 12-33%), 5 of 66, 8% (95%CI: 5-22%), and 50 of 66, 76% (95%CI: 64-85%), respectively. Antibodies to hepatitis C virus (anti-HCV) prevalence was 57 of 66, 86% (95%CI: 76-94%). No human immunodeficiency virus (HIV) positive case was found. Prevalence of past herpes simplex virus type 2 (HSV-2) infection was 29% (95%CI: 18-41%). Two patients (3%, 95%CI: 0-10%) were positive for Treponema pallidum and 18% (95%CI: 10-30%) were human herpesvirus 8 (HHV-8) antibody positive. Four hundred and fifty-two subjects were recruited for comparison. Markers of past HAV infection was associated with haemodialysis (Fisher s exact test p-value=0.037). Dialysed patients were at a higher risk of being HBsAg positive than others: the sex- and age-adjusted odds ratio (OR) was 5.18 (95%CI: 1.87-14.32). Anti-HBc positivity was strongly associated with haemodialysis: the sex- and age-adjusted OR was 6.43 (95%CI: 3.22-12-85). Anti-HCV positivity was 86% and 1% in presence and absence of haemodialysis, respectively. The Fisher s exact test for association proved a strong association between haemodialysis and HCV (p value<0.0001). The OR for association between haemodialysis and HSV-2 positivity was 3.20 (95%CI: 1.46-7.00). Significant associations were also observed between haemodialysis status and antibodies to Treponema pallidum (Fisher s exact test p value=0.044). In Kosovo, the prevalence of viral hepatitis infection and other viral infections and Treponema pallidum among dialysed patients is high, indicating major ongoing nosocomial transmission. PMID- 20003899 TI - HIV/STI co-infection among men who have sex with men in Spain. AB - In Spain, neither the HIV nor the STI national surveillance systems collect information on HIV/STI co-infection. However, there are two networks based on HIV/STI clinics which gather this data. We describe HIV prevalence in men who have sex with men (MSM) diagnosed with infectious syphilis and/or gonorrhoea in 15 STI clinics; and concurrent diagnoses of STI in MSM newly diagnosed with HIV in 19 HIV/STI clinics. In total, 572 MSM were diagnosed with infectious syphilis and 580 with gonorrhoea during 2005-2007. HIV prevalence among syphilis and gonorrhoea cases was 29.8% and 15.2% respectively. In the multivariate analysis, HIV/syphilis co-infection was associated with being Latin American; having a history of STI; reporting exclusively anal intercourse; and having sex with casual or several types of partners. HIV and gonorrhoea co-infection was associated with age older than 45 years; having no education or only primary education completed; and having a history of STI. In total, 1,462 HIV infections were newly diagnosed among MSM during 2003-2007. Of these, 31.0% were diagnosed with other STI at the same time. Factors associated with STI co-infection among new HIV cases in MSM were being Latin American; and having sex with casual partners or with both steady and casual partners. In Spain, a considerable proportion of MSM are co-infected with HIV and STI. PMID- 20003904 TI - Single-nucleotide polymorphism in the SCCmec-orfX junction distinguishes between livestock-associated MRSA CC398 and human epidemic MRSA strains. AB - A number of real-time PCR assays for direct detection of methicillinresistant (MRSA) in clinical specimens are targeting staphylococcal cassette chromosome mec (SCCmec) right extremity sequences and the S. aureus chromosomal orfX gene sequences located to the right of the SCCmec integration site. When testing 184 MRSA strains of human and animal origin from geographically distinct locations, we identified several characteristic single-nucleotide polymorphisms (SNPs) within the SCCmec-orfX junction of livestock-associated (LA) MRSA CC398 which serve as suitable strain markers for screening purposes. Within an assay time of 60 minutes and an additional 10 minutes for the melting curve analysis, all MRSA CC398 isolates were correctly identified by their characteristic T(m) value in the commercial LightCycler MRSA Advanced test. Studies to confirm the diagnostic accuracy of the SNP-based strain identification assay with a larger collection of clinical and LA-MRSA strains are ongoing. PMID- 20003906 TI - Prolonged shedding of influenza A(H1N1)v virus: two case reports from France 2009. AB - We observed a prolonged shedding of virus 14 and 28 days after symptom onset in two patients with pandemic H1N1 influenza, who did not have immunodepression and were treated with neuraminidase inhibitor. This prolonged shedding was not associated with the emergence of resistance mutation H275Y in the viral neuraminidase gene. PMID- 20003905 TI - Detection of human norovirus from frozen raspberries in a cluster of gastroenteritis outbreaks. AB - We describe a cluster of norovirus outbreaks affecting about 200 people in Southern Finland in September and October 2009. All outbreaks occurred after consumption of imported raspberries from the same batch intended for the catering sector. Human norovirus genotype GI.4 was found in frozen raspberries. The berries were served in toppings of cakes in separate catering settings or mixed in curd cheese as a snack for children in a daycare center. The relative risk for consumption of the berry dish was 3.0 (p 10.5, p=0.003. There was no difference in the mean acute antibody titers for patients with mild compared to severe illness. Serum antibody may play a role in protection from hMPV infection supporting the development of an hMPV vaccine that stimulates humoral immunity. PMID- 20003920 TI - Duration of Vi antibodies in participants vaccinated with Typhim Vi (Typhoid Vi polysaccharide vaccine) in an area not endemic for typhoid fever. AB - After a single injection of Typhim Vi (typhoid Vi polysaccharide vaccine), serum antibody concentrations were monitored for 3 years in 37 adults who resided where typhoid fever was not endemic. Anti-Vi antibody concentrations declined progressively during the study, to levels that support the current US recommendation for revaccination every 2 years. PMID- 20003921 TI - Stimulation of anti-polio and anti-HSV IgA pre-plasma cell response in blood following parenteral immunization with tetanus-diphtheria vaccine. AB - Systemic immunization can elicit a significant response of IgG producing activated B cell subsets in human blood, part of which is not toward the vaccine. However, the effect of vaccination on IgA antibody secreting B cell subsets has had limited investigation. We immunized healthy, adult volunteers with a tetanus/diphtheria vaccine and observed a significant burst of IgA-secreting pre plasma cells (PPC). Isolated PPC produced IgG, but not IgA antibody to tetanus antigen, and produced IgA and IgG antibody specific for poliovirus and herpes simplex virus. Thus, a vaccine that generates a systemic recall response to tetanus also induces blood PPC secreting IgA and IgG antibody relevant to mucosal protection. PMID- 20003922 TI - An analysis of the Human Papilloma Virus vaccine debate on MySpace blogs. AB - BACKGROUND: The roll out of HPV immunization programs across the United States was hindered by controversy. We tracked the debate in the United States through MySpace, then the most popular social networking site, in order to better understand the public's reaction to the vaccine. METHODS: We searched MySpace for all blog discourse related to HPV immunization. We analyzed each blog according to the overall portrayal of HPV immunization, identified the characteristics of the bloggers, and developed a content analysis to categorize the types of supporting arguments made. RESULTS: 303 blogs met our inclusion criteria. 157 (52%) of the blogs were classified as positive, 129 (43%) as negative, and 17 (6%) were ambivalent toward HPV immunization. Positive blogs generally argued that HPV infection was effective and there were no reasonable alternatives to immunizing. Negative blogs focused on the risks of immunizing and relied heavily on vaccine-critical publications to support their viewpoint. Of the blogs where gender could be identified, 75 (25%) were posted by men and 214 (71%) by women. 60% of blogs posted by men were explicitly critical about HPV immunization versus 36% of women's blogs. Male bloggers also had larger networks of friends. CONCLUSIONS: We describe a novel and promising approach to the surveillance of public opinions and attitudes toward immunization. In our analysis, men were far more likely to hold negative views about HPV immunization than women and disseminate negative messages through larger social networks. Blog analysis is a useful tool for Public health officials to profile vaccine criticism and to design appropriate educational information tailored to respond to alternative media/alternative information actively disseminated via social media tools. Public health officials should examine mechanisms by which to leverage this media to better communicate their message through existing networks and to engage in on going dialogue with the public. PMID- 20003923 TI - Virus-like particles and capsomeres are potent vaccines against cutaneous alpha HPVs. AB - The potential as prophylactic vaccines of L1-based particles from cutaneous genus alpha human papillomavirus (HPV) types has not been assessed so far. However, there is a high medical need for such vaccines since HPV-induced skin warts represent a major burden for children and for immunocompromised adults, such as organ transplant recipients. In this study, we have examined the immunogenicity of capsomeres and virus-like particles (VLPs) from HPV types 2, 27, and 57, the most frequent causative agents of skin warts. Immunization of mice induced immune responses resembling those observed upon vaccination with HPV 16 L1-based antigens. The antibody responses were cross-reactive but type-restricted in their neutralizing capacities. Application of adjuvant led to an enhanced potential to neutralize the respective immunogen type but did not improve cross neutralization. Vaccination with capsomeres and VLPs from all four analyzed HPV types induced robust IFNgamma-associated T-cell activation. Immunization with mixed VLPs from HPV types 2, 27, and 57 triggered an antibody response similar to that after single-type immunization and capable of efficiently neutralizing all three types. Our results imply that vaccination with combinations of VLPs from cutaneous HPV types constitutes a promising strategy to prevent HPV-induced skin lesions. PMID- 20003924 TI - Impact of influenza-like illness and effectiveness of influenza vaccination in oncohematological children who have completed cancer therapy. AB - In order to evaluate the impact of influenza-like illness and the effectiveness of influenza vaccination in children with oncohematological disease who have completed cancer therapy, 182 children with a diagnosis of oncohematological disease were divided into two subgroups on the basis of the length of time off therapy (<6 months or 6-24 months) and randomised 1:1 to receive influenza vaccination or not. The controls were 91 otherwise healthy children unvaccinated against influenza. The results show that the clinical and socioeconomic impact of influenza-like illnesses and the effectiveness of influenza vaccination in oncohematological children who have completed cancer therapy are related to the length of the off therapy period, and seem to be significantly greater in those who have been off therapy for less than 6 months in comparison with healthy controls. This suggests that the administration of influenza vaccination should be strongly recommended only among oncohematological children who have been off therapy for less than 6 months. PMID- 20003925 TI - Retro-engineering of liposomal vaccine adjuvants: role of a microarray-based screen. PMID- 20003926 TI - Efficacy of live attenuated influenza vaccine in children against influenza B viruses by lineage and antigenic similarity. AB - Seasonal influenza vaccines, including live attenuated influenza vaccine (LAIV), contain three vaccine strains (two type A and one type B). Ideally, the hemagglutinin antigens of the recommended vaccine strains are antigenically similar to epidemic wild-type strains; in actuality, the antigenic match between circulating and vaccine strains each year can vary significantly owing to intermittent genetic reassortment and continuous antigenic drift. For influenza B, antigenic relatedness is further complicated by the existence of two distinct lineages. Consequently, the influenza B vaccine component can be of a completely different antigenic lineage from the circulating epidemic strains. Using data from nine randomized clinical trials in young children (6 months to 6 years of age), vaccine efficacy of LAIV against influenza B strains was assessed across this spectrum of antigenic relatedness. In an integrated analysis, vaccine efficacy of two doses of LAIV in vaccine-naive children was 86% against B strains of the same lineage and closely matched to the vaccine strain, 55% against strains of the same lineage but antigenically drifted from the vaccine strain, and 31% against strains of the opposite B lineage and antigenically unrelated to the vaccine strain. These data provide a more accurate assessment of the protection provided by the current trivalent vaccine and highlight the need for vaccination strategies that provide enhanced protection against both lineages of influenza B such as a quadrivalent influenza vaccine. PMID- 20003927 TI - A systematic review of measures for reducing injection pain during adult immunization. AB - OBJECTIVE: To evaluate the effectiveness of different pain-relieving interventions to reduce pain from immunization in adults. DATA SOURCES: MEDLINE (1950 to October Week 3 2008) PsycINFO (1967 to December Week 1 2008), CINAHL (1982 to October Week 4 2008), EMBASE (1980 to 2008 Week 43) and the Cochrane Central Register of Controlled Trials (3rd Quarter 2008). REVIEW METHODS: Databases were searched for trials of pharmacological, behavioural, physical or operator-dependant techniques to reduce pain from immunization in adults. The primary outcome was pain as assessed by visual analogue scale or other numeric rating scale. RESULTS: Six studies representing 853 participants were identified. One study evaluating pharmacological interventions (lidocaine-prilocaine) found them to be effective in reducing pain from immunization. Similarly, two studies evaluating physical pain relieving techniques, either skin cooling interventions (Fluori-Methane) or tactile stimulation (manual pressure at the site of injection) found them to reduce pain. One study of jet injectors found them to be more painful than conventional needle and syringe. Neither freezing needles nor warming vaccines was found to be effective in reducing pain. No studies investigated psychological interventions or oral analgesics (acetaminophen and ibuprofen). CONCLUSION: There was limited evidence to support the use of lidocaine-prilocaine, Fluori-Methane and manual pressure for reducing immunization pain in adults. There was limited evidence of more pain with jet injectors compared to needle and syringe. Due to limited data, we recommend further investigation of methods to reduce immunization pain in adults, primarily psychological and physical techniques. PMID- 20003928 TI - 7-Valent pneumococcal conjugate vaccine and lower respiratory tract infections: effectiveness of a 2-dose versus 3-dose primary series. AB - BACKGROUND: Immunogenicity studies suggest antibody responses from a 7-valent pneumococcal conjugate vaccine (PCV7) regimen consisting of 2 doses in the primary series are less immunogenic, for at least several vaccine serotypes, compared with a regimen consisting of 3 doses; evidence of effectiveness for prevention of invasive pneumococcal disease for both regimens is available but comparative data are lacking for prevention of lower respiratory tract diseases (LRTD). METHODS: We compared rates of LRTD between children who were born in 2002 and received 2 versus 3 PCV7 doses in the primary series, both before and after receipt of the booster dose, using a retrospective matched-cohort design and health insurance claims data. Two-dose and 3-dose children were matched (1:1) using propensity scoring. Cumulative rates of hospital admissions and outpatient visits for LRTD were tallied during the post-primary/pre-booster period and the post-booster period (to age 3 years), respectively. RESULTS: During the post primary/pre-booster period, 3-dose children (n=3293) had 7.8 (95% CI: 0.8 to 14.8) fewer LRTD-related hospital admissions (per 1000 children) and 57 (95% CI: 6 to 128) fewer LRTD-related outpatient visits (per 1000 children) than matched 2 dose subjects (n=3293). During the post-booster period, the numbers of LRTD related hospital admissions and outpatient visits did not differ significantly between 3-dose and 2-dose children. CONCLUSIONS: Our findings suggest that a 2 dose PCV7 primary series, while conferring savings from reduced vaccine costs in comparison with a 3-dose primary series, also may confer less protection against LRTD in the first year of life, at least during the period soon after the vaccine is introduced. PMID- 20003929 TI - Fate of the tissue in lateral canals and apical ramifications in response to pathologic conditions and treatment procedures. AB - INTRODUCTION: This article reviews and reports on the histopathologic and histobacteriologic status of the tissue in lateral canals and apical ramifications (LC/AR) in diverse clinical conditions as well as in response to endodontic treatment. METHODS: In total, serial sections from 493 human tooth specimens obtained by extraction or apical surgery were screened for the presence of LC/AR. RESULTS: LC/AR were observed in about 75% of the teeth. In clinically vital teeth, vital tissue was consistently found in LC/AR. In teeth with periodontal disease, the whole pulp became necrotic only when the subgingival biofilm reached the main apical foramen. In teeth with pulp exposure by caries, the tissue in LC/AR remained vital as far as the pulp tissue in the main canal did so. When pulp necrosis reached the level of the LC/AR, the tissue therein was either partially or completely necrotic. Chemomechanical preparation partially removed necrotic tissue from the entrance of LC/AR, whereas the adjacent tissue remained inflamed, sometimes infected, and associated with periradicular disease. Vital tissue in LC/AR was not removed by preparation. In cases in which lateral canals appeared radiographically "filled," they were actually not obturated, and the remaining tissue in the ramification was inflamed and enmeshed with the filling material. CONCLUSIONS: Overall, the belief that lateral canals must be injected with filling material to enhance treatment outcome was not supported by literature review or by our histopathologic observations. It appears that strategies other than finding a technique that better squeezes sealer or gutta percha within LC/AR should be pursued to effectively disinfect these regions. PMID- 20003930 TI - Mineral trioxide aggregate: a comprehensive literature review--Part I: chemical, physical, and antibacterial properties. AB - INTRODUCTION: An ideal orthograde or retrograde filling material should seal the pathways of communication between the root canal system and its surrounding tissues. It should also be nontoxic, noncarcinogenic, nongenotoxic, biocompatible, insoluble in tissue fluids, and dimensionally stable. Mineral trioxide aggregate (MTA) was developed and recommended initially because existing root-end filling materials did not have these "ideal" characteristics. MTA has also been recommended for pulp capping, pulpotomy, apical barrier formation in teeth with open apexes, repair of root perforations, and root canal filling. Since MTA's introduction in 1993, numerous studies have been published regarding various aspects of this material. The aim of Part I of this literature review is to present investigations regarding the chemical, physical, and antibacterial properties of MTA. METHODS: A review of the literature was performed by using electronic and hand-searching methods for the chemical and physical properties and antibacterial activity of MTA from November 1993-September 2009. RESULTS: There are many published reports regarding the chemical, physical, and antibacterial properties of MTA. Our search showed that MTA is composed of calcium, silica, and bismuth. It has a long setting time, high pH, and low compressive strength. It possesses some antibacterial and antifungal properties, depending on its powder-to-liquid ratio. CONCLUSIONS: MTA is a bioactive material that influences its surrounding environment. PMID- 20003931 TI - Treatment outcome in endodontics: the Toronto study--phases 3, 4, and 5: apical surgery. AB - INTRODUCTION: The long-term outcome of apical surgery performed on root-filled teeth presenting with post-treatment apical periodontitis has been the subject of debate; therefore, current evidence is required to support the prognosis of this important procedure. The objectives of this study were (1) to assess the long term outcome of apical surgery and (2) to identify significant outcome predictors in Phases 3-5 of the Toronto Study, pooled with the previously reported Phases 1 and 2. METHODS: The 4- to 10-year outcome of apical surgery was prospectively assessed by a blinded, independent, calibrated examiner and dichotomized as "healed" (periapical index score 45 years, 84%, 10 mm, 53%). CONCLUSIONS: In this 4- to 10-year cohort study, the outcome was better in subjects >45 years old, teeth with inadequate root-filling length, and crypt size of middle > apical. CONCLUSIONS: The reducing agent was capable of reversing the effect of dentin deproteinization, and RelyX U100 behaved similarly to RelyX ARC. PMID- 20003952 TI - Effect of various chemicals as solvents on the dissolution of set white mineral trioxide aggregate: an in vitro study. AB - INTRODUCTION: The main disadvantages of mineral trioxide aggregate (MTA) are its prolonged setting time and the difficulty in retrieving it from the root canal. Because MTA is alkaline in nature, it might interact with acids or chemicals and could disintegrate. The aim of this study was to assess the effect of various chemicals on dissolution of white MTA (WMTA). METHODS: Stainless steel molds were prepared and filled with WMTA that was exposed to carbonic acid, 2% chlorhexidine gluconate solution, 17% ethylenediaminetetraacetic acid (EDTA) solution, and saline on days 1 and 21. The surface hardness was measured before and after 5-, 10-, 15-, and 20-minute intervals after exposure to chemicals. The samples were probed with a #16 endodontic explorer after 20 minutes of chemical exposure and hardness testing. RESULTS: Both qualitative and quantitative analysis showed that carbonic acid was effective in significantly reducing the surface hardness of set WMTA on both days 1 and 21; 2% chlorhexidine gluconate reduced the surface hardness of set WMTA significantly on day 1 only. CONCLUSIONS: Carbonic acid can be effectively used as an adjunct to dissolve set WMTA even after 21 days, whereas 2% chlorhexidine gluconate showed significant surface dissolution only within 24 hours of WMTA placement. Thus, usage of chlorhexidine gluconate as a root canal irrigant in which WMTA is used during endodontic procedure should be avoided for 24 hours. Solutions with EDTA have no effect on surface hardness of WMTA. PMID- 20003953 TI - Percentages of gutta-percha-filled canal area observed after increased apical enlargement. AB - INTRODUCTION: Increased apical enlargement might be achieved with S-ApeX files. However, S-ApeX preparations exhibit a small taper, which might hinder the performance of the root filling. The aim of this study was to compare the percentage of gutta-percha-filled canal areas (PGCA) in root canals after preparation with GT and S-ApeX instruments. METHODS: Four groups of maxillary lateral incisors (n = 20) were root canal treated. Canals were prepared with S ApeX files to size 60 in 2 groups or with a GT system to size 30 taper 0.06 in other groups. Canals were filled with gutta-percha and AH26 by using a lateral compaction (LC) technique or a combined lateral and vertical compaction (LCVC) technique. Roots were then sectioned 2, 4, 6, and 8 mm from the apex. The area of the canal and gutta-percha was measured in each section, and the PGCA was calculated. RESULTS: The overall PGCA in the S-ApeX/LC group was significantly lower than that in other groups (analysis of variance, P = .001). The average PGCA values were 97.6%, 99.3%, and 98.5% for the GT/LC, S-ApeX/LCVC, and GT/LCVC groups, respectively. The differences between these groups were not statistically significant (analysis of variance, P = .21). CONCLUSIONS: A combined lateral and vertical compaction technique achieved comparable PGCA in canals prepared in maxillary lateral incisors by using S-ApeX or GT instruments. PMID- 20003954 TI - Evaluation of a sonic device designed to activate irrigant in the root canal. AB - INTRODUCTION: The aims of this study were to evaluate the removal of dentin debris from the root canal by sonic or ultrasonic activation of the irrigant and the physical mechanisms of sonic activation by visualizing the oscillations of the sonic tip, both inside and outside the confinement of the root canal. METHODS: Roots of 18 canines were embedded, split, and prepared into standardized root canals. A standard groove was cut on the wall of one half of each root canal and filled with the same amount of dentin debris before irrigation procedures. The removal of dentin debris was evaluated after different irrigation procedures. The oscillations of the sonic tip were visualized ex vivo by using high-speed imaging at a time scale relevant to the irrigation process, and the oscillation amplitude of the tip was determined under 20x magnification. RESULTS: After irrigation, there was a statistically significant difference between the experimental groups (P < .0001). Without irrigant activation, the grooves were still full of dentin debris. From the ultrasonic activated group, 89% of the canals were completely free of dentin debris, whereas from the sonic group, 5.5% 6.7% were (P = .0001). There was no significant difference between the sonic activation groups. CONCLUSIONS: Activation of the irrigant resulted in significantly more dentin debris removal; ultrasonic activation was significantly more efficient than sonic activation. The oscillation amplitude of the sonically driven tips is 1.2 +/- 0.1 mm, resulting in much wall contact and no cavitation of the irrigant. PMID- 20003955 TI - Cyclic fatigue and fracture characteristics of ground and twisted nickel-titanium rotary files. AB - INTRODUCTION: The purposes of this study were to compare the fatigue resistance of traditional, ground nickel-titanium rotary instruments with the Twisted File and to examine the fracture characteristics of the fatigued fragment. METHODS: Size #25, 0.06 tapered, TF (SybronEndo), RaCe (FKG Dentaire), Helix (DiaDent), and ProTaper F1 (Dentsply Maillefer) were examined with scanning electron microscope for surface characteristics before subjected to a cyclic (rotational bending) fatigue test. The time until fracture was recorded to calculate the number of revolutions for each instrument. The data were compared for differences by using analysis of variance and post hoc Scheffe test. The fragments were examined with scanning electron microscope both in lateral view and fractographically. RESULTS: TF showed a significantly higher resistance to cyclic fatigue than other nickel-titanium files that were manufactured with a grinding process (P < .05). The path of crack propagation appeared to be different for electropolished (TF and RaCe) versus non-electropolished (Helix and ProTaper) instruments. CONCLUSIONS: Although all specimens showed similar fractographic appearance, which indicated a similar fracture mechanism, instruments with abundant machining grooves seemed to have a higher risk of fatigue. PMID- 20003956 TI - Fracture resistance and deflection of pulpless anterior teeth restored with composite or porcelain veneers. AB - INTRODUCTION: The aim of this in vitro study was to evaluate the influence that resin composite and porcelain veneer restorations, associated or not to fiber post placement, have on fracture resistance and deflection of pulpless anterior teeth. METHODS: One hundred twenty freshly extracted human maxillary central incisors were selected. Teeth were randomly divided into 7 experimental groups (veneer preparation/resin composite veneer placement/endodontic therapy and resin composite veneer placement/endodontic therapy, fiber post and resin composite veneer placement/porcelain veneer placement/endodontic therapy and porcelain veneer placement/endodontic therapy, fiber post and porcelain veneer placement) and a control group (n = 15). Specimens were loaded to fracture recording crown deflection, and data were statistically analyzed. RESULTS: Veneer preparations did not significantly influence fracture resistance of incisors. On the contrary, veneer preparation significantly increased specimen deflection values. Fiber posts seemed to significantly increase mean maximum load values for endodontically treated teeth restored with either composite or porcelain veneers. CONCLUSIONS: A fiber post restoration can be suggested when endodontic treatment is associated with veneer restoration. Veneer restorations seem to be an optimal choice also for endodontically treated teeth. PMID- 20003957 TI - Alternating versus continuous rotation: a comparative study of the effect on instrument life. AB - INTRODUCTION: Rotary nickel-titanium instruments have multiple advantages but with the risk of more fractures because of flexure or torsion. Alternating rotation appears to be an alternative to continuous rotary movement. The objective of this study was to determine the influence of the type of instrument rotation on the frequency of fractures or deformation. METHODS: Instrumentation was performed on 120 molar root canals with an angle of curvature greater than 30 degrees using alternating rotation (group A: 60 degrees clockwise, 45 degrees counterclockwise) and continuous rotation (group B). RESULTS: The results indicate that instruments used with alternating rotation have a higher mean number of uses (13.0) compared with the continuous rotation group (10.05); this difference was statistically significant (p < 0.05). CONCLUSIONS: The ProTaper shaping instruments (S1 and S2) are those that achieved the greatest difference in use with alternating rotation, with S2 being the most resistant to fracture or deformation with the two types of movement used. PMID- 20003958 TI - Use of mineral trioxide aggregate in the treatment of invasive cervical resorption: a case report. AB - INTRODUCTION: Guided tissue regeneration (GTR) is a successful treatment procedure of periodontal reconstructive surgery. Adverse effects can be observed after GTR procedure, but external root resorption is rarely reported at clinical studies. Invasive cervical resorption is a clinical term used to describe a relatively uncommon, insidious, and often aggressive form of external root resorption. METHODS: The present case demonstrates an invasive cervical resorption that is a potential late complication of GTR in a maxillary canine of a 59-year-old male patient. After the surgical intervention and root canal treatment, the resorption was subsequently sealed with mineral trioxide aggregate. RESULTS: The 1-year follow-up demonstrates no pathologic changes on clinical and radiographic examination. CONCLUSIONS: This case report presents a treatment strategy that might improve the healing outcomes for patients with invasive cervical resorption. PMID- 20003959 TI - Management of mucosal fenestrations by interdisciplinary approach: a report of three cases. AB - INTRODUCTION: Mucosal fenestrations are infrequently encountered in clinical practice, and as such, their management has not been reported often. Their treatment might be further complicated by communication, with the oral environment making them susceptible to the deposition of plaque and calculus, a secondary factor in preventing reformation of mucosal covering. METHODS: Three cases of mucosal fenestrations are presented, of which 2 were nonvital and were treated endodontically. Full-thickness flap was elevated; root-end resection and root-end filling with light-cured glass ionomer were done. In the third case, the tooth was vital, so the fenestrated root surface was just planed. The areas were then covered with connective tissue graft before repositioning the flap. RESULTS: At subsequent follow-up, the mucosal fenestrations were completely healed by soft tissue coverage, and postoperative radiographs revealed good periapical bone healing in the first 2 cases, and there was maintenance of vitality in the third case. CONCLUSIONS: Careful diagnosis of mucosal fenestrations can help elucidate the underlying cause, and combining nonsurgical and surgical therapies can be a good treatment option for their successful management. PMID- 20003960 TI - [Cerebral aneurysm and subarachnoid hemorrhage revealed by a Tako Tsubo syndrome]. AB - Cerebral hemorrhage is usually associated to many cardiac disorders, mimicking acute coronary syndrome. We relate a case of a postmenopausal woman presenting at emergency room for acute coronary syndrome and whose evaluation revealed a subarachnoidal hemorrhage, normal coronary arteries, and a typical Tako Tsubo aspect on echocardiography with apical ballooning and hyperkinesis with basal hypokinesis. PMID- 20003961 TI - [Iatrogenic dissection of the right coronary artery and the ascending aorta during coronary intervention]. AB - Iatrogenic acute dissection of the ascending aorta following coronary angiography and percutaneous intervention is rare. The options for treatment are dictated by patient stability, nature of dissection of the coronary vessel, ability to restore the coronary circulation and extent of aortic dissection. Usually localized aortic dissections have been managed conservatively or treated by sealing the entry with a coronary stent. Extensive dissections may require a surgical intervention. We report the case of a 52-year-old man with iatrogenic dissection of the right coronary artery ostium and extension of the dissection to the ascending aorta during intraluminal angioplasty of an obstructive lesion in the first portion of the right coronary artery. The patient was managed conservatively without stenting (failure stenting of the right coronary artery) and without surgery. Aortic dissection was monitored by means of transesophageal echocardiography. Serial computed tomography scans demonstrated spontaneous resolution of the dissection. The evolution of the patient was satisfactory. Causes, frequency and treatment procedures of this iatrogeny are discussed. PMID- 20003962 TI - Parasympathectomy induces morphological changes and alters gene-expression profiles in the rat submandibular gland. AB - OBJECTIVE: The chorda-lingual (CL) nerve carries parasympathetic fibers to the hilum of the sublingual and submandibular glands (SMGs) and evokes the secretion of saliva. The effect of cutting the CL nerve on the biological processes in SMGs was investigated by examining the gene-expression profiles in the SMGs after a surgical parasympathectomy. METHODS: At day 3 after the CL nerve cut, the changes in the SMGs at both the experimental cut and contralateral control sides were analysed by microarray and light microscopy. The expression levels of 6 selected genes were confirmed by real-time PCR, Western blot and immunofluorescence staining. RESULTS: The wet weight of the parasympathectomised SMGs decreased significantly compared to that of the contralateral side (p<0.05). Histological analyses after the parasympathectomy showed a widened interacinar space as well as some atropic changes to the acini of the SMGs in the cut side. Microarray analysis revealed that twofold differential expression in mRNA expression in the parasympathectomized SMGs were detected in 88 genes (0.004%): 41 genes were overexpressed, 11 were underexpressed and 36 were unknown. Changes of the expression of 6 selected genes detected by Western blot and/or real-time PCR were consistent with the microarray data. CONCLUSION: The important genes involved in biological processes for salivation were identified through a large-scale gene expression analysis. PMID- 20003964 TI - Response time distributional evidence for distinct varieties of number attraction. AB - Speakers are known to make subject-verb agreement errors both when a number mismatching noun intervenes between the head of the subject phrase and the verb (e.g., *The key to the cabinets are on the table) and in configurations in which there is a number-mismatching noun that does not intervene (e.g., *The cabinets that the key open are on the second floor). Using a two-choice response time (RT) paradigm, Staub (2009) found that correct agreement decisions were also slowed in both cases. The present article reports a new experiment designed to explore whether these two RT effects are qualitatively similar or different. Fitting of the ex-Gaussian distribution (Ratcliff, 1979) to individual subjects' RT data, in each condition, demonstrated that the effect of an intervening number attractor on correct RT is due to both a shifting of the distribution to the right and to increased skewing, while the effect of a non-intervening attractor is almost entirely a skewing effect. A non-parametric vincentizing procedure supported these conclusions. These findings are taken to support the view that these two types of number attraction involve distinct processing mechanisms. PMID- 20003963 TI - Novel O-antigen of Hafnia alvei PCM 1195 lipopolysaccharide with a teichoic acid like structure. AB - The lipopolysaccharide (LPS) of Hafnia alvei strain PCM 1195 was obtained by the hot phenol/water method. The O-specific polysaccharide was released by mild acidic hydrolysis and isolated by gel filtration. The structure of the O-specific polysaccharide was investigated by (1)H, (13)C, and (31)P NMR spectroscopy, MALDI TOF MS, and GC-MS, accompanied by monosaccharide and methylation analysis. It was concluded that the O-specific polysaccharide is composed of a hexasaccharide repeating units interlinked with a phosphate group: {-->4-alpha-D-Glcp-(1-->3) alpha-L-FucpNAc-(1-->3)-[alpha-D-Glcp-(1-->4)]-alpha-D-GlcpNAc-(1-->3)-alpha-L FucpNAc-(1-->4)-alpha-D-Glcp-(1-->P}(n). PMID- 20003965 TI - Pesticide exposure of pregnant women in Guadeloupe: ability of a food frequency questionnaire to estimate blood concentration of chlordecone. AB - CONTEXT: Chlordecone, an environmentally persistent organochlorine insecticide used intensively in banana culture in the French West Indies until 1993, has permanently polluted soils and contaminated foodstuffs. Consumption of contaminated food is the main source of exposure nowadays. We sought to identify main contributors to blood chlordecone concentration (BCC) and to validate an exposure indicator based on food intakes. MATERIAL AND METHODS: We used a food frequency questionnaire (FFQ) completed by a sample of 194 pregnant women to estimate their dietary exposure to chlordecone and compared it to blood levels. In a first approach, chlordecone daily intake was estimated as the product of daily eaten quantity of 214 foodstuffs, multiplied by their chlordecone content, and summed over all items. We then predicted individual blood chlordecone concentration with empirical weight regression models based on frequency of food consumption, and without contamination data. RESULTS: Among the 191 subjects who had BCC determination, 146 (76%) had detectable values and mean BCC was 0.86 ng/mL (range < LOD-13.2). Mean per capita dietary intake of chlordecone was estimated at 3.3 microg/day (range: 0.1-22.2). Blood chlordecone levels were significantly correlated with food exposure predicted from the empirical weight models (r=0.47, p<0.0001) and, to a lesser extent, with chlordecone intake estimated from food consumption and food contamination data (r=0.20, p=0.007). Main contributors to chlordecone exposure included seafood, root vegetables, and Cucurbitaceous. CONCLUSION: These results show that the Timoun FFQ provides valid estimates of chlordecone exposure. Estimates from empirical weight models correlated better with blood levels of chlordecone than did estimates from the dietary intake assessment. PMID- 20003966 TI - Surgical gastrojejunostomy or endoscopic stent placement for the palliation of malignant gastric outlet obstruction (SUSTENT study): a multicenter randomized trial. AB - BACKGROUND: Both gastrojejunostomy (GJJ) and stent placement are commonly used palliative treatments of obstructive symptoms caused by malignant gastric outlet obstruction (GOO). OBJECTIVE: Compare GJJ and stent placement. DESIGN: Multicenter, randomized trial. SETTING: Twenty-one centers in The Netherlands. PATIENTS: Patients with GOO. INTERVENTIONS: GJJ and stent placement. MAIN OUTCOME MEASUREMENTS: Outcomes were medical effects, quality of life, and costs. Analysis was by intent to treat. RESULTS: Eighteen patients were randomized to GJJ and 21 to stent placement. Food intake improved more rapidly after stent placement than after GJJ (GOO Scoring System score > or = 2: median 5 vs 8 days, respectively; P < .01) but long-term relief was better after GJJ, with more patients living more days with a GOO Scoring System score of 2 or more than after stent placement (72 vs 50 days, respectively; P = .05). More major complications (stent: 6 in 4 patients vs GJJ: 0; P = .02), recurrent obstructive symptoms (stent: 8 in 5 patients vs GJJ: 1 in 1 patient; P = .02), and reinterventions (stent: 10 in 7 patients vs GJJ: 2 in 2 patients; P < .01) were observed after stent placement compared with GJJ. When stent obstruction was not regarded as a major complication, no differences in complications were found (P = .4). There were also no differences in median survival (stent: 56 days vs GJJ: 78 days) and quality of life. Mean total costs of GJJ were higher compared with stent placement ($16,535 vs $11,720, respectively; P = .049 [comparing medians]). Because of the small study population, only initial hospital costs would have been statistically significant if the Bonferroni correction for multiple testing had been applied. LIMITATIONS: Relatively small patient population. CONCLUSIONS: Despite slow initial symptom improvement, GJJ was associated with better long term results and is therefore the treatment of choice in patients with a life expectancy of 2 months or longer. Because stent placement was associated with better short-term outcomes, this treatment is preferable for patients expected to live less than 2 months. ( CLINICAL TRIAL REGISTRATION NUMBER: ISRCTN 06702358.). PMID- 20003967 TI - A new metal stent with a controlled-release system for palliation of malignant dysphagia: a prospective, multicenter study. AB - BACKGROUND: Self-expanding metal stents are frequently used for the palliation of malignant dysphagia. Recently, a new stent design, the Evolution stent (Cook Medical, Limerick, Ireland), was developed. The stent has an internal and external silicone coating to resist tissue ingrowth and uncoated dual flanges on both ends to prevent migration. Its controlled-release system may allow full control and precise positioning of the stent. OBJECTIVE: To determine the safety and clinical effectiveness of the Evolution stent for the palliation of dysphagia in patients with malignant strictures located in the esophagus (n = 33 [75%]) or gastric cardia (n = 11 [25%]). METHODS: All patients who underwent placement of an esophageal Evolution stent from February to December 2008 received follow-up telephone calls 14 days after treatment and thereafter monthly until death. Data on technical and clinical outcome, complications, and recurrent dysphagia were collected. RESULTS: A total of 44 patients (32 [73%] men, median age 74 years [range 48-91 years]) were treated with the Evolution stent. The median tumor length was 6 cm (range 1-11 cm). Stent placement was technically successful in 41 patients. Dysphagia improved from a median of 3 (ability to swallow liquids only) to 1 (ability to eat some solid foods) 4 weeks after stent placement. Major complications occurred in 4 (9%) patients, ie, pneumonia (3 patients) and hemorrhage (1 patient). Two (5%) patients died of pneumonia. Minor complications included gastroesophageal reflux and mild retrosternal pain in 11 (25%) and 4 (9%) patients, respectively. Recurrent dysphagia developed in 11 (25%) patients as a result of tissue ingrowth or overgrowth (6 patients; 14%), food impaction (3 patients; 7%), or stent migration (2 patients; 5%). CONCLUSIONS: The Evolution stent is safe and effective for the palliation of dysphagia from esophageal and gastric cardia cancer. Randomized trials are needed to compare this device with other stent designs. PMID- 20003968 TI - Endoscopic peritoneal access and insufflation: natural orifice transluminal endoscopic surgery. AB - BACKGROUND: Diagnostic transgastric endoscopic peritoneoscopy is a safe model for exploration of the peritoneum. Endoscopic insufflation of the peritoneal cavity has not been validated in humans. We report here our experience with pneumoperitoneum established endoscopically with a laparoscopic insufflator. DESIGN: Pneumoperitoneum was established with a laparoscopic insufflator through the biopsy channel of the gastroscope. Intra-abdominal pressure was measured with a transfascial Veress needle and compared with endoscopic values. The gastrotomy was used in the creation of the gastric pouch. PATIENTS: Twenty patients undergoing laparoscopic Roux-en-Y gastric bypass participated in the study. Ten had undergone no previous surgery, whereas the other 10 patients had a history of abdominal procedures. INTERVENTIONS: Diagnostic transgastric endoscopic peritoneoscopy was performed through a gastrotomy created endoscopically without laparoscopic visualization. MAIN OUTCOME MEASUREMENTS: Diagnostic findings, operating times, and clinical course were recorded. RESULTS: The average time for transgastric access was 9.6 minutes. This did not vary in patients with previous surgery (P = .3). Endoscopic insufflation was successful in all patients. The mean endoscopic and laparoscopic pressures were 9.80 and 9.75 mm Hg, respectively (P = .9). In no patients were there limitations to visualization of the abdomen. Adhesions were noted in 80% and 10% of patients with and without a history of surgery, respectively (P = .005). There were no complications related to transgastric passage of the endoscope or exploration of the peritoneal cavity. CONCLUSIONS: Although limited by the small sample size in this study, we believe that transgastric access may be considered as an alternative approach to peritoneal insufflation and provides a safe alternative for exploration of the abdomen. Endoscopic insufflation through the biopsy channel by using a laparoscopic insufflator seems to be an effective and safe method for establishing pneumoperitoneum. PMID- 20003969 TI - Needle-knife sphincterotomy: factors predicting its use and the relationship with post-ERCP pancreatitis (with video). AB - BACKGROUND: In the absence of precut needle-knife sphincterotomy (NKS), failure of biliary cannulation may occur in up to 10% of cases. There are few prospective evaluations of the safety and efficacy of NKS, and studies of its early use in difficult cannulation have been inconclusive. Whether precut NKS after failure of primary biliary cannulation is independently associated with post-ERCP pancreatitis (PEP) remains controversial. OBJECTIVE: To examine the relationship between NKS and PEP. DESIGN: Analysis of prospectively collected data from two randomized trials of ERCP techniques, with PEP as the primary endpoint measure. SETTING: Tertiary-care academic medical center. PATIENTS: This study involved 732 patients from two successive, prospective, randomized trials of naive papilla cannulation between November 2001 and April 2006. Patients with pancreatic or ampullary cancer were excluded. INTERVENTION: Naive papilla cannulation, NKS, primary guidewire versus contrast-assisted cannulation, and glyceryl trinitrate patch versus placebo. MAIN OUTCOME MEASUREMENTS: PEP and procedure-related complications. RESULTS: NKS was performed in 94 of 732 patients (12.8%) and was successful in achieving bile duct access in 80 of 94 patients (85%). Cannulation success in the entire group was 717 of 732 patients (97.7%). The overall frequency of PEP following NKS was 14.9% (14 of 94 patients) compared with 6.1% (39 of 638 patients) without NKS (P < .001). The incidence of PEP increased with an increasing number of attempts at cannulating the papilla. Pancreatic stents were inserted in 22 patients, 5 of whom developed pancreatitis. In multivariate analysis, independent predictors of PEP were as follow: female sex (odds ratio [OR] = 3.5, P = .028), suspected sphincter of Oddi dysfunction (SOD) (OR = 9.7, P < .001), partial pancreatic drainage (OR = 4.8, P = .011), 10 to 14 attempts at papilla cannulation (OR = 4.4, P = .031), and >/=15 attempts at papilla cannulation (OR = 9.4, P = .013). NKS was not an independent predictor of PEP. There were no perforations, no major bleeding, and no cases of severe pancreatitis in the NKS group. LIMITATIONS: Nonrandomized for NKS intervention. CONCLUSIONS: The number of attempts at cannulating the papilla is independently associated with PEP, and the risk increases with an increasing number of attempts. NKS is not an independent predictor of PEP. PMID- 20003970 TI - A new fully covered stent with antimigration properties for the palliation of malignant dysphagia: a prospective cohort study. AB - BACKGROUND: Fully covered stents are designed to resist tissue ingrowth that is often seen with partially covered stents. An issue with fully covered stents is the risk of migration. OBJECTIVE: We aimed to determine efficacy, recurrent dysphagia, and complications of the SX-ELLA stent Esophageal HV, which is fully covered to resist tissue ingrowth and has an antimigration ring to withstand migration. DESIGN: Prospective cohort study. SETTING: Two tertiary referral centers. PATIENTS: Forty-four patients with malignant esophageal strictures from inoperable or metastatic esophageal or gastric cardia cancer (n = 42) or lung cancer (n = 2). INTERVENTIONS: Placement of an SX-ELLA stent. MAIN OUTCOME MEASURES: Functional outcome, recurrent dysphagia, complications, and survival. RESULTS: Dysphagia improved from a median score of 3 (liquids only) before stent placement to 1 (ability to eat some solid food) 4 weeks later (P < .001). Twelve of 44 (Kaplan Meier analysis = 40%) patients developed 18 episodes of recurrent dysphagia of which 6 were caused by stent migration and 2 by tissue overgrowth. In total, 14 episodes of major complications developed in 10 of 44 (Kaplan Meier analysis = 29%) patients, 8 of which were caused by hemorrhage. After a median follow-up of 15 months, 39 patients had died (median survival 110 days), 5 (11%) from hemorrhage. LIMITATIONS: Nonrandomized study design. CONCLUSIONS: Dysphagia caused by esophageal cancer can be successfully palliated by placement of a new, fully covered esophageal stent (SX-ELLA). Although this single-wire braided stent with an antimigration ring is supposed to be less traumatic and to reduce migration, this was not substantiated in this study. Further improvements of stent features are needed to achieve the goals set for this study. PMID- 20003971 TI - Does Barrett's esophagus respond to chemoradiation therapy for adenocarcinoma of the esophagus? AB - BACKGROUND: Adenocarcinoma of the esophagus is frequently associated with Barrett's esophagus (BE). The response of esophageal adenocarcinoma to chemoradiation therapy is well described; however, the effect of chemoradiation on tumor-associated BE has not been specifically reported. OBJECTIVE: To determine the response of tumor-associated BE to chemoradiation therapy. DESIGN: Retrospective cohort study. SETTING: A single National Cancer Institute Comprehensive Cancer Care Center experience. PATIENTS: The study cohort consisted of 43 patients with stage I to IVA esophageal adenocarcinoma associated with BE who received either neoadjuvant or definitive chemoradiation therapy and underwent either esophagectomy or surveillance at our institution. MAIN OUTCOME MEASUREMENT: The presence and extent of BE after chemoradiation therapy of esophageal adenocarcinoma associated with endoscopically documented pretreatment BE. RESULTS: BE persisted after chemoradiation therapy in 93% (40/43) of cases (95% CI, 83%-99%). Twenty-seven patients received neoadjuvant chemoradiation therapy before esophagectomy. Persistent BE was detected in all 27 surgical specimens (100%). In 59% (16/27) of the cases, there was complete pathologic tumor response. Sixteen patients received definitive chemoradiation therapy. Persistent pretreatment BE was identified in 88% (14/16) by surveillance endoscopy (95% CI, 60%-98%). The mean length of BE before and after chemoradiation was 6.6 cm and 5.8 cm, respectively (P = .38). LIMITATIONS: Retrospective design, small sample size, and single-site data collection. CONCLUSIONS: Chemoradiation therapy of esophageal adenocarcinoma does not eliminate tumor-associated BE, nor does it affect the length of the BE segment. PMID- 20003972 TI - Time-gated fluorescence spectroscopy improves endoscopic detection of low-grade dysplasia in ulcerative colitis. AB - BACKGROUND: Dysplasia in ulcerative colitis is frequently missed with 4-quadrant biopsies. An experimental setup recording delayed fluorescence spectra simultaneously with white light endoscopy was recently developed. OBJECTIVE: We compared detection of invisible flat intraepithelial neoplasia with protoporphyrin IX fluorescence and standard 4-quadrant biopsies. DESIGN: Prospective, crossover design without randomization of the order of procedures. SETTING: Gastroenterology Department, Humboldt University, Charite, Berlin, Germany. PATIENTS: Forty-two patients with extensive ulcerative colitis of more than 10 years' duration were included. INTERVENTIONS: Colonoscopy with 4-quadrant biopsies and targeted biopsies of macroscopic lesions and time-gated fluorescence guided colonoscopy were performed 2 weeks apart by 2 blinded endoscopists. Three independent pathologists examined the biopsy specimens. MAIN OUTCOME MEASUREMENTS: The primary outcome criterion was detection rate of invisible flat intraepithelial neoplasia. RESULTS: Invisible flat intraepithelial neoplasia was detected in 3 (7%) patients by white light 4-quadrant biopsies and in 10 (24%) patients by fluorescence-guided endoscopy (P = .02). The sensitivity and specificity for differentiating patients with and without dysplasia were 100% and 81%, respectively. Dysplastic and nondysplastic mucosa could be discriminated with a sensitivity and specificity of 73% and 81%, respectively. LIMITATIONS: The trial was not randomized. CONCLUSION: The detection rate of intraepithelial neoplasia in patients with ulcerative colitis can be improved by fluorescence guided colonoscopy. PMID- 20003973 TI - Massive abdominal venous cavernous transformation diagnosed by use of EUS. PMID- 20003974 TI - Stabilization of nonlinear systems with a slowly varying parameter by a control Lyapunov function. AB - Based on a control Lyapunov function (CLF) strategy, a novel approach for designing a controller for a slowly varying nonlinear system is proposed. The approach may be thought of as being in between the time-invariant and time varying CLF techniques. If the time-invariant technique is used to control a slowly varying system, stability will not be guaranteed. On the other hand, the time-varying CLF technique, due to the control law, has complexity and needs to measure or estimate the derivative of system parameters. The advantage of the proposed method is its independence from the measurement or estimation of the derivatives of the system parameters. It is shown that the proposed control law can even be independent of the parameters of the system. In this paper, the conditions are derived that allow using the simple CLF formula that guarantees the stability of a slowly varying system. The efficiency of the approach is shown through some simulations. PMID- 20003975 TI - Pressure ulcer risk assessment in critical care: interrater reliability and validity studies of the Braden and Waterlow scales and subjective ratings in two intensive care units. AB - BACKGROUND: The application of standardized pressure ulcer risk assessment scales is recommended in clinical practice. OBJECTIVES: The aims of this study were to compare the interrater reliabilities of the Braden and Waterlow scores and subjective pressure ulcer risk assessment and to determine the construct validity of these three assessment approaches. DESIGN: Observational. SETTINGS: Two intensive care units of a large University Hospital in Germany. PARTICIPANTS: 21 and 24 patients were assessed by 53 nurses. Patients' mean age was 69.7 (SD 8.3) and 67.2 (SD 11.3). METHODS: Two interrater reliability studies were conducted. Samples of patients were assessed independently by a sample of three nurses. A 10 cm visual analogue scale was applied to measure subjective pressure ulcer risk rating. Intraclass correlation coefficients (ICC) and standard errors of measurement (SEM) were used to determine interrater reliability and agreement of the item and sum scores. Pearson product moment correlation coefficients (r) were used to indicate the degree and direction of the relationships between the measures. RESULTS: The interrater reliability for the subjective pressure ulcer risk assessment was ICC(1,1)=0.51 (95% CI 0.26-0.74) and 0.71 (95% CI 0.53-0.85). Interrater reliability of Braden scale sum scores was ICC(1,1)=0.72 (95% CI 0.52 0.87) and 0.84 (95% CI 0.72-0.92) and for Waterlow scale sum scores ICC(1,1)=0.36 (95% CI 0.09-0.63) and 0.51 (95% CI 0.27-0.72). The absolute degree of correlation between the measures ranged from 0.51 to 0.77. CONCLUSIONS: Interrater reliability coefficients indicate a high degree of measurement error inherent in the scores. Compared to subjective risk assessment and the Waterlow scale scores the Braden scale performed best. However, measurement error is too high to draw valid inferences for individuals. Less than 26-59% of variances in scores of one scale were determined by scores of another scale indicating that all three instruments only partly measured the same construct. The use of the Braden-, Waterlow- and Visual Analogue scales for measuring pressure ulcer risk of intensive care unit patients is not recommended. PMID- 20003976 TI - Interplay of genetic and environmental factors: Innate immunity genetic polymorphisms in MBL2 affect endothelial dysfunction and risk of atherosclerosis. PMID- 20003977 TI - Effect of transglutaminase 2 (TG2) deficiency on atherosclerotic plaque stability in the apolipoprotein E deficient mouse. AB - BACKGROUND: Transglutaminase 2 (TG2), a cross-linking enzyme that confers supra molecular structures with extra rigidity and resistance against proteolytic degradation, is expressed in the shoulder regions of human atherosclerotic plaques. It has been proposed that TG2 prevents tearing and promotes plaque repair at these potential weak points, and also promotes ectopic calcification of arteries. TG2 is also expressed within plaques that develop within the brachiocephalic arteries of apolipoprotein E (apoE) deficient mice. OBJECTIVES: To determine the role that TG2 plays in plaque development and calcification, mice were bred that were doubly deficient in apoE and TG2, and were maintained on a high-fat diet for 6 months. RESULTS: Lesion size and composition were not significantly altered in the apoE/TG2 double-knockout mice, with the exception of a 9.7% decrease in the proportion of the plaque occupied by lipid (p=0.032). The frequency of buried fibrous caps within brachiocephalic plaques was significantly higher in male than in female mice, but TG2 deficiency had no effect on either gender. The extent of lesion calcification varied markedly between individual mice, but it was not decreased in the apoE/TG2 double-knockout mice. CONCLUSION: These data indicate that, in the apoE knockout mouse model of atherosclerosis, TG2 does not influence plaque composition or calcification. The data further suggest that TG2 does not influence plaque stability or repair in these mice. PMID- 20003978 TI - Contact mechanics and elastohydrodynamic lubrication in a novel metal-on-metal hip implant with an aspherical bearing surface. AB - Diameter and diametral clearance of the bearing surfaces of metal-on-metal hip implants and structural supports have been recognised as key factors to reduce the dry contact and hydrodynamic pressures and improve lubrication performance. On the other hand, application of aspherical bearing surfaces can also significantly affect the contact mechanics and lubrication performance by changing the radius of the curvature of a bearing surface and consequently improving the conformity between the head and the cup. In this study, a novel metal-on-metal hip implant employing a specific aspherical bearing surface, Alpharabola, as the acetabular surface was investigated for both contact mechanics and elastohydrodynamic lubrication under steady-state conditions. When compared with conventional spherical bearing surfaces, a more uniform pressure distribution and a thicker lubricant film thickness within the loaded conjunction were predicted for this novel Alpharabola hip implant. The effects of the geometric parameters of this novel acetabular surface on the pressure distribution and lubricant thickness were investigated. A significant increase in the predicted lubricant film thickness and a significant decrease in the dry contact and hydrodynamic pressures were found with appropriate combinations of these geometric parameters, compared with the spherical bearing surface. PMID- 20003980 TI - Stress relaxation of human ankles is only minimally affected by knee and ankle angle. AB - Comprehensive characterization of stress relaxation in musculotendinous structures is needed to create robust models of viscoelastic behavior. The commonly used quasi-linear viscoelastic (QLV) theory requires that the relaxation response be independent of tissue strain (length). This study aims to characterize stress relaxation in the musculotendinous and ligamentous structures crossing the human ankle (ankle-only structures and the gastrocnemius muscle tendon unit, which crosses the ankle and knee), and to determine whether stress relaxation is independent of the length of these structures. Two experiments were conducted on 8 healthy subjects. The first experiment compared stress relaxation over 10 min at different gastrocnemius muscle-tendon unit lengths keeping the length of ankle-joint only structures fixed. The second experiment compared stress relaxation at different lengths of ankle-joint only structures keeping gastrocnemius muscle-tendon unit length fixed. Stress relaxation data were fitted with a two-term exponential function (T=G(0)+G(1)e(-lambda(1))(t)+G(2)e( lambda(2))(t)). The first experiment demonstrated a significant effect of gastrocnemius muscle-tendon unit length on G(1), and the second experiment demonstrated an effect of the length of ankle-joint only structures on G(2), lambda(1) and lambda(2) (p<0.05). Nonetheless, the size of effects on stress relaxation was small (DeltaG/G<10%), similar to experimental variability. We conclude that stress relaxation in the relaxed human ankle is minimally affected by changing gastrocnemius muscle-tendon unit length or by changing the lengths of ankle-joint only structures. Consequently quasi-linear viscoelastic models of the relaxed human ankle can use a common stress relaxation modulus at different knee and ankle angles with minimal error. PMID- 20003979 TI - Compressive axial mechanical properties of rat bone as functions of bone volume fraction, apparent density and micro-ct based mineral density. AB - Mechanical testing has been regarded as the gold standard to investigate the effects of pathologies on the structure-function properties of the skeleton. With recent advances in computing power of personal computers, virtual alternatives to mechanical testing are gaining acceptance and use. We have previously introduced such a technique called structural rigidity analysis to assess mechanical strength of skeletal tissue with defects. The application of this technique is predicated upon the use of relationships defining the strength of bone as a function of its density for a given loading mode. We are to apply this technique in rat models to assess their compressive skeletal response subjected to a host of biological and pharmaceutical stimulations. Therefore, the aim of this study is to derive a relationship expressing axial compressive mechanical properties of rat cortical and cancellous bone as a function of equivalent bone mineral density, bone volume fraction or apparent density over a range of normal and pathologic bones. We used bones from normal, ovariectomized and partially nephrectomized animals. All specimens underwent micro-computed tomographic imaging to assess bone morphometric and densitometric indices and uniaxial compression to failure. We obtained univariate relationships describing 71-78% of the mechanical properties of rat cortical and cancellous bone based on equivalent mineral density, bone volume fraction or apparent density over a wide range of density and common skeletal pathologies. The relationships reported in this study can be used in the structural rigidity analysis introduced by the authors to provide a non-invasive method to assess the compressive strength of bones affected by pathology and/or treatment options. PMID- 20003981 TI - Features of the adsorption of Naproxen on the chiral stationary phase (S,S)-Whelk O1 under reversed-phase conditions. AB - Using elution chromatography, we studied the adsorption mechanism of the Naproxen enantiomers on the chiral stationary phase (S,S)-Whelk-O1, from buffered methanol water solutions. We propose an adsorption mechanism that assumes monolayer adsorption of the more retained enantiomer and the associative adsorption of the less retained one. The effects of the mobile phase composition on the adsorption of Naproxen are discussed. The combination of an elevated column temperature and of the use of an acidic mobile phase led to the degradation of the column and caused a major loss of its separation ability. The use of a moderately acidic mobile phase at temperature slightly above ambient did not produce rapid severe damages but, nevertheless, hampered the experiments and caused a slow gradual deterioration of the column. PMID- 20003982 TI - Editorial on "The challenges of the analysis of basic compounds by high performance liquid chromatography: some possible approaches for improved separations" by D.V. McCalley. PMID- 20003983 TI - Simultaneous quantification of multiple classes of phenolic compounds in blood plasma by liquid chromatography-electrospray tandem mass spectrometry. AB - A method using high performance liquid chromatography-electrospray tandem mass spectrometry (LC-ESI-MS/MS) in positive ion mode was developed for the simultaneous analysis of 30 phenolic compounds, including four estrogens, bisphenol A (BPA), 10 hydroxylated polybrominated dephenyl ethers (OH-PBDEs) and 15 bromophenols (BRPs), in blood plasma. In the present method, derivatization with dansyl chloride was employed, and all the derivertized target compounds were well resolved on a 100mm Xbridge C18 column with acetonitrile and 0.1% formic acid as the mobile phases. Purification procedures, such as liquid-liquid extraction and silica-gel chromatography, were applied to reduce matrix effects in the sample extract and remove excess derivatizing reagents, thus permitting selective and sensitive detection of the target phenolic compounds. The limit of quantification for all analytes, with a signal-to-noise ratio of approximately 10, was 2-30 pg/g (plasma weight) except for 6-OH-BDE-137 (30 pg/g) and 3-BRP (60 pg/g). The method was validated for recoveries (68-100%), accuracy (84-110%) and precision (3.7-11%) using charcoal-stripped bovine blood plasma spiked with all target compounds (500 and 5000 pg/mL). Finally, the method was applied to analyze six blood plasma samples from frogs and cormorants, where two natural estrogens, one BPA, one OH-PBDE and four BRPs were detected. The greatest total concentrations of estrogens coincided with the least total concentrations of other phenolic compounds for both species. The proposed method based on derivatization followed by LC-MS/MS provides a novel method to simultaneously monitor multiple groups of phenolic compounds in blood plasma. PMID- 20003984 TI - Synergistic effect of silicate clay and phosphazene-oxyalkyleneamines on thermal stability of cured epoxies. AB - Amine substitution of hexachlorocyclophosphazene (HCP) with poly(oxypropylene)diamines (POP) afforded HCP-POP adducts which were subsequently intercalated into a layered silicate clay. The relative thermal stabilities of the epoxies cured with the phosphazene-amines and the intercalated clays were studied. The organoclays, with the confined HCP-POP from 400 and 2000 g/mol M(w) amines, are nongelled products when using 1/6 M ratio of HCP/POP starting materials in tetrahydrofuran solvent. The intercalation of HCP-POP polyamine salts into sodium montmorillonite afforded the HCP-POP-embedded organoclays with an expanded interlayer silicate spacing (2.4-5.1 nm) from the original 1.2 nm spacing (X-ray diffraction). The effect of silicate clays was evaluated by blending the HCP-POP/clay hybrids into a two-component epoxy system (diglycidyl ether of 4,4'-isopropylidenediphenol (BPA) and a diamine) and fully cured to form solid materials. The distribution of the exfoliated silicate platelets in the matrix was analyzed by transmission electronic microscopy (TEM). Thermal gravimetric analysis (TGA) indicated an enhanced thermal stability for the HCP/clay epoxy nanocomposites, with a delayed weight-loss pattern (temperature of weight loss at 10% (T(10 wt.%)) from 360 to 385 degrees C and temperature of weight loss at 85% (T(85 wt.%)) from 598 to 696 degrees C), compared to the pristine epoxies. By comparing these epoxies with different amounts of phosphazene and/or silicates, the TGA revealed a synergistic effect for the presence of both phosphorous and silicate components. Furthermore, the epoxies had improved physical properties such as hardness (from 3H to 5H) and surface adhesion (observed by scanning electron microscope (SEM) on fracture surface). PMID- 20003985 TI - High adsorption selectivity of ZnAl layered double hydroxides and the calcined materials toward phosphate. AB - The selective adsorption of phosphate ions was investigated on CO(3)(2-), Cl(-), and NO(3)(-)-type ZnAl layered double hydroxides (LDHs) and the calcined CO(3)(2 )-type LDH at 200, 400, 600 and 800 degrees C, respectively. The calcined LDHs and NO(3)(-)-type LDH showed high selectivity toward phosphate ions, while the Cl(-)-type LDH shows selectivity toward both SO(4)(2-) and phosphate ions. The NO(3)(-)-type LDH selectively adsorbed phosphate ions mainly through ion exchange. The calcined samples possibly proceed through ligand complexation or electrostatic attraction between phosphate ions and hydrated ZnO formed after calcination, although the structural memory effect was observed for the samples calcined below 600 degrees C with the formation of CO(3)(2-)-type LDH. Adsorption/desorption behaviors indicated that the calcined samples had good stability and reusability. PMID- 20003986 TI - Beta-lactoglobulin aggregates in foam films: effect of the concentration and size of the protein aggregates. AB - Single foam films made from mixtures of nonaggregated proteins and protein aggregates have been studied using a thin film balance apparatus. Their features (heterogeneity, stability and resistance to pressure change) are dependent on the aggregate size and on the ratio between nonaggregated proteins and protein aggregates. A phase diagram of these foam films has been drawn and a correlation of the structural properties of foam film and stability of real foams has been found. In particular, the formation of a gel-like network within the foam film coincides with the stability of the corresponding 3-D foams. PMID- 20003987 TI - The effect of age on the phenotype and function of developing thymocytes. AB - The immune system declines with age leading to a progressive deterioration in the ability to respond to infection and vaccination. Age-associated thymic involution is one of the most recognized changes in the ageing immune system and is believed to be a major contributor towards immunosenescence; however, the precise mechanisms involved in age-associated thymic involution remain unclear. In order to gain further insight into the effect of ageing on T-cell development, steady state thymopoiesis was studied in mice ranging from 1 to 18 months of age. There was a decrease in thymic cellularity with age, but the most dramatic loss occurred early in life. Although there were no alterations in the proportion of the major thymocyte subsets, there was a significant decline in the expression of other key molecules including CD3 and CD24. There was a decline in the ability of thymocytes from older mice to respond to mitogens, which was demonstrated by a failure to up-regulate expression of the activation marker CD69 and to enter the G(2)--M phase of the cell cycle. This was concurrent with an increased resistance to apoptosis in thymocytes from aged animals. Together, these results suggest that T cells may be flawed even before exiting to the periphery and that this could contribute to the age-associated decline in immune function. PMID- 20003988 TI - Reading development in agglutinative languages: evidence from beginning, intermediate, and adult Basque readers. AB - Do typological properties of language, such as agglutination (i.e., the morphological process of adding affixes to the lexeme of a word), have an impact on the development of visual word recognition? To answer this question, we carried out an experiment in which beginning, intermediate, and adult Basque readers (n=32 each, average age=7, 11, and 22 years, respectively) needed to read correctly versus incorrectly inflected words embedded in sentences. Half of the targets contained high-frequency stems, and the other half contained low frequency stems. To each stem, four inflections of different lengths were attached (-a, -ari, -aren, and -arentzat, i.e., inflectional sequences). To test whether the process of word recognition was modulated by the knowledge of word structure in the language, half of the participants' native language was Basque and the other half's native language was Spanish. Children showed robust effects of frequency and length of inflection that diminished with age. In addition, the effect of length of inflection was modulated by the frequency of the stem and by the native language. Taken together, these results suggest that word recognition develops from a decoding strategy to a direct lexical access strategy and that this process is modulated by children's knowledge of the inflectional structure of words from the beginning of their reading experience. PMID- 20003989 TI - Early-life stress and antidepressant treatment involve synaptic signaling and Erk kinases in a gene-environment model of depression. AB - Stress has been shown to interact with genetic vulnerability in pathogenesis of psychiatric disorders. Here we investigated the outcome of interaction between genetic vulnerability and early-life stress, by employing a rodent model that combines an inherited trait of vulnerability in Flinders Sensitive Line (FSL) rats, with early-life stress (maternal separation). Basal differences in synaptic signaling between FSL rats and their controls were studied, as well as the consequences of early-life stress in adulthood, and their response to chronic antidepressant treatment (escitalopram). FSL rats showed basal differences in the activation of synapsin I and Erk1/2, as well as in alpha CaM kinase II/syntaxin-1 and alpha CaM kinase II/NMDA-receptor interactions in purified hippocampal synaptosomes. In addition, FSL rats displayed a blunted response of Erk-MAP kinases and other differences in the outcome of early-life stress in adulthood. Escitalopram treatment restored some but not all alterations observed in FSL rats after early-life stress. The marked alterations found in key regulators of presynaptic release/neurotransmission in the basal FSL rats, and as a result of early-life stress, suggest synaptic dysfunction. These results show that early gene-environment interaction may cause life-long synaptic changes affecting the course of depressive-like behavior and response to drugs. PMID- 20003990 TI - A novel approach to prevent spinal cord ischemia: Inoue stent graft with a side branch of small caliber for the reconstruction of the artery of Adamkiewicz. AB - OBJECTIVES: Paraplegia remains a serious complication after endovascular repair of thoracic aortic aneurysms, and it has been reported that paraplegia might be due to ischemia of the artery of Adamkiewicz. This study investigates the feasibility of an Inoue stent graft with a side branch of a small caliber for the reconstruction of the intercostal artery branching the artery of Adamkiewicz. METHODS: Branched Inoue stent grafts were implanted into the thoracic aorta and 11th intercostal artery of 5 mongrel dogs. The side branch measured 3 x 5 mm and contained a bare-metal coronary stent for fixing to the intercostal arterial wall. Aortography and selective angiography of the 11th intercostal artery were performed before and immediately after implantation and after 1, 4, 8, and 12 weeks. The luminal diameter of the intercostal artery before implantation was 2.4 +/- 0.3 mm. RESULTS: All stent grafts were successfully deployed. The main body of the graft did not develop endoleak or migrate, and the side branch remained patent for 12 weeks. Angiography performed 1 week postoperatively revealed smooth flow with slight stenosis (4.2% +/- 1.7%) along the side branches and the intercostal arteries in all dogs. Four weeks postoperatively, however, mild concentric stenosis (38% +/- 16%) along the side branch was observed in 4 of the 5 dogs. The percent stenosis at 8 and 12 weeks was 38% +/- 15% and 33% +/- 11%, respectively; these values were not significantly different from the value at 4 postoperative weeks. CONCLUSIONS: A novel Inoue stent graft with a side branch of small caliber was successfully deployed into the canine thoracic aorta and intercostal artery; the side branch remained patent for 12 weeks. This novel technique may enable endovascular reconstruction of the Adamkiewicz artery. PMID- 20003991 TI - The Arctic is no longer put on ice: evaluation of Polar cod (Boreogadus saida) as a monitoring species of oil pollution in cold waters. AB - The withdrawing Arctic ice edge will facilitate future sea transport and exploration activities in the area, which calls for the establishment of relevant cold water monitoring species. The present study presents first results of field baseline levels for core oil pollution biomarkers in Polar cod (Boreogadussaida) sampled from pristine, Arctic waters. Furthermore, biomarker response levels were characterized in controlled laboratory exposure experiments running over 2 weeks. Fish exposed to a simulated petrogenic spill (1ppm dispersed, crude oil) exhibited elevated hepatic EROD activity, bile PAH-metabolites, and hepatic DNA adducts, whereas male individuals exposed to simulated produced water (30ppb nonylphenol) exhibited a strong induction of plasma vitellogenin. In conclusion, the results demonstrated low and robust biomarker baseline levels that were clearly different from exposure responses. In combination with its high abundance and circumpolar distribution, the Polar cod seems well qualified for oil pollution monitoring in Arctic waters. PMID- 20003992 TI - Blast furnace slag can effectively remediate coastal marine sediments affected by organic enrichment. AB - There is an urgent need to control nutrient release fluxes from organically enriched sediments into overlying waters to alleviate the effects of eutrophication. This study aims to characterize blast furnace slag (BFS) and evaluate its remediation performance on organically-enriched sediments in terms of suppressing nutrient fluxes and reducing acid volatile sulfide. BFS was mainly composed of inorganic substances such as CaO, SiO(2), Al(2)O(3) and MgO in amorphous crystal phase. Container experiments showed that the phosphate concentration in the overlying water, its releasing flux from sediment and AVS of the sediment decreased by 17-23%, 39% and 16% compared to the control without BFS, respectively. The loss on ignition was significantly decreased by 3.6-11% compared to the control. Thus, the application of BFS to organically-enriched sediment has a suppressive role on organic matter, AVS concentration and phosphate releasing flux from sediments and therefore, is a good candidate as an effective environmental remediation agent. PMID- 20003993 TI - Association of secular trends in unemployment with suicide in Taiwan, 1959-2007: a time-series analysis. AB - OBJECTIVE: Despite the wealth of research investigating the association of unemployment with suicide in the West, few studies have investigated the association in non-Western countries. This study aimed to investigate the relationship between secular trends in unemployment and suicide in Taiwan. STUDY DESIGN: Time-series analysis. METHODS: Overall and age-specific suicide rates (1959-2007) for Taiwanese men and women aged 15 years or above were calculated from national population and mortality statistics. The association of secular trends in unemployment with suicide was investigated graphically and using time series modelling (Prais-Winsten regression). RESULTS: Rises in unemployment were associated with an increase in male suicide rates, but evidence for an association in females was limited. In the model controlling for changes in gross domestic product (GDP) per capita, GDP growth, divorce and female labour force participation, for every 1% rise in unemployment, male suicide rates increased by 3.1 (95% confidence interval 1.4-4.8) per 100,000. There is no evidence for a difference in the strength of association between men of different ages. CONCLUSION: Trends in suicide, particularly for adult males, appear to be influenced by unemployment. The results have implications for suicide prevention, in particular for societies facing acute rises in unemployment during recessions. PMID- 20003994 TI - Developing adenoviral vectors encoding therapeutic genes toxic to host cells: comparing binary and single-inducible vectors expressing truncated E2F-1. AB - Adenoviral vectors are highly efficient at transferring genes into cells and are broadly used in cancer gene therapy. However, many therapeutic genes are toxic to vector host cells and thus inhibit vector production. The truncated form of E2F-1 (E2Ftr), which lacks the transactivation domain, can significantly induce cancer cell apoptosis, but is also toxic to HEK-293 cells and inhibits adenovirus replication. To overcome this, we have developed binary- and single-vector systems with a modified tetracycline-off inducible promoter to control E2Ftr expression. We compared several vectors and found that the structure of expression cassettes in vectors significantly affects E2Ftr expression. One construct expresses high levels of inducible E2Ftr and efficiently causes apoptotic cancer cell death by activation of caspase-3. The approach developed in this study may be applied in other viral vectors for encoding therapeutic genes that are toxic to their host cells and/or inhibit vector propagation. PMID- 20003996 TI - Formation and fate of chlorination by-products in reverse osmosis desalination systems. AB - Chlorination by-products may be formed during pretreatment or posttreatment disinfection in reverse osmosis (RO) desalination systems, potentially posing health, aesthetic and ecological risks. To assess the formation and fate of by products under different conditions likely to be encountered in desalination systems, trihalomethanes, dihaloacetonitriles, haloacetic acids, and bromophenols were analyzed in water samples from a pilot-scale seawater desalination plant with a chlorine pretreatment system and in benchscale experiments designed to simulate other feed water conditions. In the pilot plant, RO rejection performance as low as 55% was observed for neutral, low-molecular-weight by products such as chloroform or bromochloroacetonitrile. Benchscale chlorination experiments, conducted on seawater from various locations indicated significant temporal and spatial variability for all by-products, which could not be explained by measured concentrations of organic carbon or bulk parameters such as SUVA(254). When desalinated water was blended with freshwater, elevated concentrations of bromide in the blended water resulted in dihaloacetonitrile concentrations that were higher than those expected from dilution. In most situations, the concentration of chlorination by-products formed from continuous chlorination of seawater or blending of desalinated water and freshwater will not compromise water quality or pose significant risks to aquatic ecosystems. PMID- 20003995 TI - A rev1-vpu polymorphism unique to HIV-1 subtype A and C strains impairs envelope glycoprotein expression from rev-vpu-env cassettes and reduces virion infectivity in pseudotyping assays. AB - Functional studies of HIV-1 envelope glycoproteins (Envs) commonly include the generation of pseudoviruses, which are produced by co-transfection of rev-vpu-env cassettes with an env-deficient provirus. Here, we describe six Env constructs from transmitted/founder HIV-1 that were defective in the pseudotyping assay, although two produced infectious virions when expressed from their cognate proviruses. All of these constructs exhibited an unusual gene arrangement in which the first exon of rev (rev1) and vpu were in the same reading frame without an intervening stop codon. Disruption of the rev1-vpu fusion gene by frameshift mutation, stop codon, or abrogation of the rev initiation codon restored pseudovirion infectivity. Introduction of the fusion gene into wildtype Env cassettes severely compromised their function. The defect was not due to altered env and rev transcription or a dominant negative effect of the expressed fusion protein, but seemed to be caused by inefficient translation at the env initiation codon. Although the rev1-vpu polymorphism affects Env expression only in vitro, it can cause problems in studies requiring Env complementation, such as analyses of co-receptor usage and neutralization properties, since 3% of subtype A, 20% of subtype C and 5% of CRF01_A/E viruses encode the fusion gene. A solution is to eliminate the rev initiation codon when amplifying rev-vpu-env cassettes since this increases Env expression irrespective of the presence of the polymorphism. PMID- 20003997 TI - Biological oxidation of dissolved methane in effluents from anaerobic reactors using a down-flow hanging sponge reactor. AB - Anaerobic wastewater treatment plants discharge dissolved methane, which is usually not recovered. To prevent emission of methane, which is a greenhouse gas, we utilized an encapsulated down-flow hanging sponge reactor as a post-treatment to biologically oxidize dissolved methane. Within 3 weeks after reactor start-up, methane removal efficiency of up to 95% was achieved with a methane removal rate of 0.8 kg COD m(-3) day(-1) at an HRT of 2 h. After increasing the methane loading rate, the maximum methane removal rate reached 2.2 kg COD m(-3) day(-1) at an HRT of 0.5 h. On the other hand, only about 10% of influent ammonium was oxidized to nitrate during the first period, but as airflow was increased to 2.5 L day(-1), nitrification efficiency increased to approximately 70%. However, the ammonia oxidation rate then decreased with an increase in the methane-loading rate. These results indicate that methane oxidation occurred preferentially over ammonium oxidation in the reactor. Cloning of the 16S rRNA and pmoA genes as well as phylogenetic and T-RFLP analyses revealed that type I methanotrophs were the dominant methane oxidizers, whereas type II methanotrophs were detected only in minor portion of the reactor. PMID- 20003998 TI - Effects of pH, ionic strength, dissolved organic matter, and flow rate on the co transport of MS2 bacteriophages with kaolinite in gravel aquifer media. AB - Viruses are often associated with colloids in wastewater and could be transported with colloids into groundwater from land disposal of human and animal effluent and sludge, causing contamination of groundwater. To investigate the role of colloids in the transport of viruses in groundwater, experiments were conducted using a 2m long column packed with heterogeneous gravel aquifer media. Bacteriophage MS2 was used as the model virus and kaolinite as the model colloid. Experimental data were analyzed using Temporal Moment Analysis and Filtration Theory. In the absence of kaolinite colloid, MS2 phage traveled slightly faster than the conservative tracer bromide (Br), with little differences observed between unfiltered and filtered MS2 phage (0.22 microm as the operational cut-off for colloid-free virus). In the presence of kaolinite colloids, MS2 phage breakthrough occurred concurrently with that of the colloidal particles and the time taken to reach the peak virus concentration was reduced, suggesting a colloid-facilitated virus transport in terms of peak-concentration time and velocity. Meanwhile mass recovery and magnitude of concentrations of the phages were significantly reduced, indicating colloid-assisted virus attenuation in terms of concentrations and mass. Decreasing the pH or increasing the ionic strength increased the level of virus attachment to the aquifer media and colloids, and virus transport became more retarded, resulting in lower peak concentration, lower mass recovery, longer peak-concentration time, and greater apparent collision efficiency. Increasing the concentration of dissolved organic matter (DOM) or flow rate resulted in faster virus transport velocity, higher peak-concentrations and mass recoveries, and lower apparent collision efficiencies. The dual-role of colloids in transport viruses has important implications for risk analysis and remediation of virus-contaminated sites. PMID- 20003999 TI - Study on the adsorption of Neutral Red from aqueous solution onto halloysite nanotubes. AB - Halloysite nanotubes (HNTs), a low-cost available clay mineral, were tested for the ability to remove cationic dye, Neutral Red (NR), from aqueous solution. Natural HNTs used as adsorbent in this work were initially characterized by XRD, FT-IR, TEM and BET. The effect of adsorbent dose, initial pH, temperature, initial concentration and contact time were investigated. Adsorption increased with increase in adsorbent dose, initial pH, temperature and initial concentration. The equilibrium data were well described by both the Langmuir and Freundlich isotherm models. The maximum adsorption capacity was 54.85, 59.24 and 65.45mg/g at 298, 308 and 318K, respectively. Batch kinetic experiments showed that the adsorption followed pseudo-second-order kinetic model with correlation coefficients greater than 0.999. Thermodynamic parameters of DeltaG(0), DeltaH(0) and DeltaS(0) indicated the adsorption process was spontaneous and endothermic. The results above confirmed that HNTs had the potential to be utilized as low cost and relatively effective adsorbent for cationic dyes removal. PMID- 20004000 TI - Pesticide exposure and inducible antipredator responses in the zooplankton grazer, Daphnia magna Straus. AB - Risk assessment of toxic substances under ecologically-relevant scenarios which include the presence of natural stressors is essential to understand their indirect toxic effects and to improve prediction of the impacts of contamination on community structure and ecosystem function. Here, we study the effects of the pesticide imidacloprid on the responses of Daphnia magna to a combination of predator-release kairomones from trout and alarm cues from conspecifics, simulating different levels of perceived predation risk. The joint effects of simultaneous exposure to both types of stressors were assessed both by traditional analysis of variance and by employing conceptual models for the evaluation of contaminant mixture exposures. Results demonstrated that pesticide exposure can significantly increase the costs of inducible antipredator defences and impair life-history responses of daphnids to fish predation pressure. Since trait-mediated effects are well-known to play a key role in population dynamics, the combined direct and indirect effects of sub-lethal concentrations of pesticides could induce maladaptive responses in zooplankton populations in the field, reducing their long-term viability. PMID- 20004001 TI - CD81 protein is expressed at high levels in normal germinal center B cells and in subtypes of human lymphomas. AB - CD81 is a tetraspanin cell surface protein that regulates CD19 expression in B lymphocytes and enables hepatitis C virus infection of human cells. Immunohistologic analysis in normal hematopoietic tissue showed strong staining for CD81 in normal germinal center B cells, a cell type in which its increased expression has not been previously recognized. High-dimensional flow cytometry analysis of normal hematopoietic tissue confirmed that among B- and T-cell subsets, germinal center B cells showed the highest level of CD81 expression. In more than 800 neoplastic tissue samples, its expression was also found in most non-Hodgkin lymphomas. Staining for CD81 was rarely seen in multiple myeloma, Hodgkin lymphoma, or myeloid leukemia. In hierarchical cluster analysis of diffuse large B-cell lymphoma, staining for CD81 was most similar to other germinal center B cell-associated markers, particularly LMO2. By flow cytometry, CD81 was expressed in diffuse large B-cell lymphoma cells independent of the presence or absence of CD10, another germinal center B-cell marker. The detection of CD81 in routine biopsy samples and its differential expression in lymphoma subtypes, particularly diffuse large B-cell lymphoma, warrant further study to assess CD81 expression and its role in the risk stratification of patients with diffuse large B-cell lymphoma. PMID- 20004002 TI - Ecological vulnerability in risk assessment--a review and perspectives. AB - This paper reviews the application of ecological vulnerability analysis in risk assessment and describes new developments in methodology. For generic non-site specific assessments (e.g. for the requirements of most European directives on dangerous chemicals) risk is characterised just on the basis of the ratio between an effect indicator and an exposure indicator. However, when the actual risk for a specific ecosystem is desired, the concept of ecological vulnerability may be more appropriate. This calls for a change in thinking, from sensitivity at the organism level to vulnerability at higher organization levels, and thus forms the link from laboratory toxicology to field effects at population, community or ecosystem level. To do so, biological and ecological characteristics of the ecosystems under concern are needed to estimate the ecological vulnerability. In this review we describe different vulnerability analysis methods developed for populations (of a single species), communities (consisting of different populations of species) and ecosystems (community and habitat combined). We also give some examples of methods developed for socio-ecological systems. Aspects that all methods share are the use of expert judgment, the input of stakeholders, ranking and mapping of the results, and the qualitative nature of the results. A new general framework is presented to guide future ecological vulnerability analysis. This framework can be used as part of ecological risk assessment, but also in risk management. We conclude that the further quantification of ecological vulnerability is a valuable contribution to vulnerability assessment. PMID- 20004003 TI - Urinary arsenic concentrations and speciation in residents living in an area with naturally contaminated soils. AB - A cross sectional study was carried out to evaluate arsenic exposure of residents living in an area with a soil naturally rich in arsenic (As), through urinary measurements. During the summer of 2007, 322 people aged over 7 years and resident in the study area for at least 4 days prior to the investigation were recruited. The sum of urinary inorganic arsenic and metabolites (iAs+MMA+DMA) and speciation were determined by graphite furnace atomic absorption spectrometry and high performance liquid chromatography coupled to inductively coupled plasma mass spectrometry, respectively. Geometric means levels of iAs+MMA+DMA were 3.6 microg/L or 4.4 microg/g creatinine. The percent of DMA, As(III) and MMA contribution to urinary arsenic concentrations was respectively 84.2%, 12% and 3.7%. We found significant associations between urinary arsenic concentrations and the consumption of seafood (p=0.03), the consumption of wine (p=0.03) and beer (p=0.001), respectively 3 and 4 days before the investigation. When we focus on the various species, As(V) was rarely detected and DMA is the predominant metabolite composing the majority of measurable inorganic-related As in the urine. Considering the percent of DMA contribution to iAs+MMA+DMA urinary concentrations, almost half of the subjects had 100% of DMA contribution whatever the concentration of urinary As whereas the others had a lower DMA contribution, between 39 and 90%. Arsenic levels reported in this original study in France were between 2 and 4 times lower than in other studies dealing with iAs+MMA+DMA levels associated with soil arsenic exposure. Arsenic levels were similar to those observed in unexposed individuals in European countries, although 10% were above the French guideline values for the general population. PMID- 20004004 TI - Polybrominated diphenyl ethers, perfluorinated alkylated substances, and metals in tile drainage and groundwater following applications of municipal biosolids to agricultural fields. AB - Polybrominated diphenyl ethers (PBDEs), perfluorinated alkylated substances (PFAS), and metals were monitored in tile drainage and groundwater following liquid (LMB) and dewatered municipal biosolid (DMB) applications to silty-clay loam agricultural field plots. LMB was applied (93,500 L ha(-1)) in late fall 2005 via surface spreading on un-tilled soil (SS(LMB)), and a one-pass aerator based pre-tillage prior to surface spreading (AerWay SSD) (A). The DMB was applied (8 Mg d wha(-1)) in early summer 2006 on the same plots by injecting DMB beneath the soil surface (DI), and surface spreading on un-tilled soil (SS(DMB)). Key PBDE congeners (BDE-47, -99, -100, -153, -154, -183, -209) comprising 97% of total PBDE in LMB, had maximum tile effluent concentrations ranging from 6 to 320 ng L(-1) during application-induced tile flow. SS(LMB) application-induced tile mass loads for these PBDE congeners were significantly higher than those for control (C) plots (no LMB) (p<0.05), but not A plots (p>0.05). PBDE mass loss via tile (0-2h post-application) as a percent of mass applied was approximately 0.04 0.1% and approximately 0.8-1.7% for A and SS(LMB), respectively. Total PBDE loading to soil via LMB and DMB application was 0.0018 and 0.02 kg total PBDE ha( 1)yr(-1), respectively. Total PBDE concentration in soil (0-0.2m) after both applications was 115 ng g(-1)dw, (sampled 599 days and 340 days post LMB and DMB applications respectively). Of all the PFAS compounds, only PFOS (max concentration=17 ng L(-1)) and PFOA (12 ng L(-1)) were found above detectable limits in tile drainage from the application plots. Mass loads of metals in tile for the LMB application-induced tile hydrograph event, and post-application concentrations of metals in groundwater, showed significant (p<0.05) land application treatment effects (SS(LMB)>A>C for tile and SS(LMB) and A>C for groundwater for most results). Following DMB application, no significant differences in metal mass loads in tile were found between SS(DMB) and DI treatments (PBDE/PFAS were not measured). But for many metals (Cu, Se, Cd, Mo, Hg and Pb) both SS(DMB) and DI loads were significantly higher than those from C, but only during <100 days post DMB application. Clearly, pre-tilling the soil (e.g., A) prior to surface application of LMB will reduce application-based PBDE and metal contamination to tile drainage and shallow groundwater. Directly injecting DMB in soil does not significantly increase metal loading to tile drains relative to SS(DMB), thus, DI should be considered a DMB land application option. PMID- 20004005 TI - Persistent organic pollutants in fish tissue in the mid-continental great rivers of the United States. AB - Great rivers of the central United States (Upper Mississippi, Missouri, and Ohio rivers) are valuable economic and cultural resources, yet until recently their ecological condition has not been well quantified. In 2004-2005, as part of the Environmental Monitoring and Assessment Program for Great River Ecosystems (EMAP GRE), we measured legacy organochlorines (OCs) (pesticides and polychlorinated biphenyls, PCBs) and emerging compounds (polybrominated diphenyl ethers, PBDEs) in whole fish to estimate human and wildlife exposure risks from fish consumption. PCBs, PBDEs, chlordane, dieldrin and dichlorodiphenyltrichloroethane (DDT) were detected in most samples across all rivers, and hexachlorobenzene was detected in most Ohio River samples. Concentrations were highest in the Ohio River, followed by the Mississippi and Missouri Rivers, respectively. Dieldrin and PCBs posed the greatest risk to humans. Their concentrations exceeded human screening values for cancer risk in 27-54% and 16-98% of river km, respectively. Chlordane exceeded wildlife risk values for kingfisher in 11-96% of river km. PBDE concentrations were highest in large fish in the Missouri and Ohio Rivers (mean>1000 ng g(-1) lipid), with congener 47 most prevalent. OC and PBDE concentrations were positively related to fish size, lipid content, trophic guild, and proximity to urban areas. Contamination of fishes by OCs is widespread among great rivers, although exposure risks appear to be more localized and limited in scope. As an indicator of ecological condition, fish tissue contamination contributes to the overall assessment of great river ecosystems in the U.S. PMID- 20004006 TI - Role of the platelet chemokine platelet factor 4 (PF4) in hemostasis and thrombosis. AB - Chemokines are a family of small proteins that have significant roles in inflammation, angiogenesis and cellular homing. Since inflammation and hemostasis/thrombosis have multiple overlapping roles and pathways, one could expect that some chemokines would also have biologically significant roles in hemostasis/thrombosis as well. This would especially be true for chemokines that are localized solely or predominantly within platelets and released in large amounts at sites of platelet activation such as platelet factor 4 (PF4, CXCL4) and its closely related chemokine, platelet basic protein (PBP, CXCL7). Our group and others have clearly demonstrated an in vivo role for PF4 in hemostasis/thrombosis, but not for PBP, which in contrast has clear proinflammatory properties. This review will focus on PF4 and its potential roles in hemostasis/thrombosis and the underlying pathways by which PF4 may be especially important in such pathologic thrombotic states as heparin-induced thrombocytopenia (HIT) and septic shock. PMID- 20004007 TI - Gamma tocopherol supplementation prevents exercise induced coagulation and platelet aggregation. PMID- 20004008 TI - Glucose as a prognostic factor in non-diabetic women with locally advanced cervical cancer (IIB-IVA). AB - OBJECTIVE: The aim of this study was to evaluate the association between pretreatment random plasma glucose levels and cancer prognosis in non-diabetic women with locally advanced cervical cancer treated with CCRT (concurrent chemoradiation) or RT (radiation therapy) only. METHODS: We investigated the non fasting plasma glucose levels checked during the initial work up before treatment in 134 non-diabetic patients with locally advanced cervical cancer. Based on the survival time and the progression-free interval (PFI) recorded in the electronic medical records Cox proportional hazard regression models were used to estimate the hazard ratio (HR) for overall survival and PFI according to the various level of glucose and a cut-off level (<102 mg/dL and >or=102 mg/dL), adjusting for clinical covariates. RESULTS: A shorter overall survival and PFI was observed in the group with higher glucose levels (HR, 1.03; p=0.002, HR, 1.02; p=0.001, respectively) and more than 102 mg/dL, by univariate analyses (HR, 3.21; p=0.012, HR, 2.20; p=0.006, respectively). Multivariate analysis, adjusting for clinical FIGO stage, performance status, treatment type (CCRT vs. RT) and chemotherapeutic regimen types showed that patients with higher glucose levels or more than >or=102 mg/dL had shorter overall survival times (HR, 1.02; p=0.015, HR, 2.54; p=0.049, respectively) and PFI (HR, 1.02; p=0.003, HR, 1.88; p=0.031, respectively). CONCLUSION: This investigation provides evidence supporting the prognostic value of glucose levels in non-diabetic women with locally advanced cervical cancer treated with radiation therapy and/or concurrent chemotherapy; high glucose levels were associated with a greater risk for recurrence and mortality in these patients. PMID- 20004009 TI - Effect of storage duration, storage temperature, and diluent on the viability and fertility of fresh ram sperm. AB - Cervical artificial insemination (AI) in sheep with fresh semen yields a much higher pregnancy rate than when frozen-thawed semen is used, and consequently frozen semen is only acceptable for laparoscopic insemination. The short life span of fresh semen is a major constraint on the use of AI in genetic improvement programs for sheep. The main objective of this study was to examine the effects of storage conditions on viability and fertilization ability of fresh ram (Ovis aries) semen up to 72h postcollection. Experiment 1 was designed to evaluate the effect of diluent type (standard skim milk, AndroMed, OviPro, and INRA 96) and storage temperature (5 degrees C and 15 degrees C) on the motility and viability of fresh ram semen. Storage temperature, irrespective of diluent, had a significant effect on both motility and viability. Storage at 5 degrees C maintained acceptable motility and viability up to 72h compared with that of storage at 15 degrees C. In Experiment 2, the penetrating ability of fresh ram semen, diluted in either skim milk, AndroMed, or INRA 96, was assessed using artificial mucus. Flat capillary tubes containing artificial mucus were suspended in 250muL semen at a sperm concentration of 20x10(6)/mL. Semen was stored at 5 degrees C and tested after 6, 24, 48, and 72h. There was a significant diluent by time interaction. In Experiment 3, the fertilizing ability of fresh ram semen stored at 5 degrees C was evaluated in vitro. Fresh semen (diluted in either skim milk, AndroMed, or INRA 96) was added to matured ewe oocytes at 6, 24, or 72h after semen collection. Cleavage rate was recorded at 48h postinsemination, and blastocyst development was recorded on Days 6 to 9. There was a significant treatment effect on cleavage and blastocyst rates; insemination of semen stored for 24h resulted in higher rates than those for storage at 72h. In Experiment 4, the fertilizing ability of fresh ram semen was evaluated in vivo. Semen was diluted in INRA 96, stored at 5 degrees C, and used to inseminate ewes on the day of collection or at 24, 48, and 72h postcollection. Multiparous ewes were cervically inseminated at a synchronized estrus. Fertility rate decreased linearly (P<0.001) up to 72h after semen collection. PMID- 20004010 TI - Effects of age, weight, hormones, and hibernation on breeding success in boreal toads (Bufo boreas boreas). AB - The goals of this study were to test the effects of exogenous hormones and hibernation on breeding behavior and gamete release by boreal toads (Bufo boreas boreas). Each year, a subset of 77 toads was hibernated and then paired with hibernated or nonhibernated mates and treated with luteinizing hormone releasing hormone analogue (LHRHa), human chorionic gonadotropin (hCG), or left untreated. Amplexus and egg and sperm production were recorded. At 1 yr of age, only 19% of pairs exhibited amplexus, and no sperm or eggs were produced. At 2 and 3 yr of age, most male toads treated with LHRHa exhibited amplexus (56.9% and 100%, respectively). Among 2-yr-old males, amplexus was more prevalent (P<0.05) in those that were hibernated than in those that were nonhibernated (54.0% and 33.3%, respectively), but most males in each group (93.3% and 75%, respectively) produced sperm in response to LHRHa treatment. Only one 2-yr-old and two 3-yr-old females produced eggs. At 4 yr of age, eight females produced eggs, but two died from egg retention. More nonhibernated than hibernated females developed eggs (7 of 10 vs. 1 of 10, P<0.05). Mean (+/-SD) weight of female toads producing eggs (58.9 +/- 11.9g) was greater (P<0.05) than that of nonproducing females (43.6 +/- 7.0g). Similarly, four of seven nonhibernated females (58.8+/-8.3g) produced eggs at 5 yr of age. All eggs were produced by females treated once with LHRHa. Number of eggs per female varied (141 to 3307), and development to tadpoles was low (0 to 36.5%), although tadpoles did become toadlets. In conclusion, male and female boreal toads matured at 2 and 4 yr of age, respectively, and heavier females were more likely to produce eggs. To enhance breeding success, males should be hibernated and treated with LHRHa. In contrast, female productivity was enhanced by improving their body condition instead of subjecting them to hibernation prior to LHRHa treatment. PMID- 20004011 TI - Postactivation treatment with nocodazole maintains normal nuclear ploidy of cloned pig embryos by increasing nuclear retention and formation of single pronucleus. AB - The objective of this study was to investigate the effects of postactivation treatment with nocodazole on morphologic changes of donor nuclei and in vitro and in vivo development of somatic cell nucleus transfer (SCNT) embryos in pigs (Sus scrofa). Somatic cell nucleus transfer oocytes were either untreated (control) or treated with nocodazole or demecolcine after electric activation, then cultured in vitro or transferred to surrogate pigs. Treatment with nocodazole (30%) and demecolcine (29%) after electric activation improved embryo development to the blastocyst stage compared with the control (16%). The rate of oocytes that formed single clusters of chromosomes or a pronucleus 4h after activation was higher after treatment with nocodazole (82%) and demecolcine (86%) than under the control conditions (66%), and this tendency was not altered even 12h after activation. Pseudo-polar body extrusion was inhibited by nocodazole and demecolcine, and the rate of embryos with diploid chromosomes was higher after treatment with nocodazole (86%) and demecolcine (77%) than under control conditions (58%). Nocodazole treatment resulted in a farrowing rate of 50% with a 1.7% efficiency of piglet production, whereas controls showed a farrowing rate of 60% and a production efficiency of 3.8%. Our results demonstrate that postactivation treatment with nocodazole maintains normal nuclear ploidy of cloned embryos likely by increasing nuclear retention and formation of single pronuclei. In vivo development could be achieved from the transfer of nocodazole treated embryos but showed some defects compared with control. PMID- 20004012 TI - Umbilical vein injection of oxytocin for retained placenta. PMID- 20004013 TI - Umbilical vein oxytocin for the treatment of retained placenta (Release Study): a double-blind, randomised controlled trial. AB - BACKGROUND: Retained placenta is associated with post-partum haemorrhage. Meta analysis has suggested that umbilical injection of oxytocin could increase placental expulsion without the need for a surgeon or anaesthetic. We assessed the effect of high-dose umbilical vein oxytocin as a treatment for retained placenta. METHODS: In this double-blind, placebo-controlled trial, haemodynamically stable women with a retained placenta for more than 30 min were recruited from 13 sites in the UK, Uganda, and Pakistan. 577 women were randomly assigned by a computer-generated randomisation list stratified by centre to 30 mL saline containing either 50 IU oxytocin (n=292) or 5 mL water (n=285), which was injected into the placenta through an umbilical vein catheter. All trial participants, study workers, and data handlers were masked to individual allocations. The primary outcome was the need for manual removal of the placenta. Analysis was by intention to treat. This study is registered, number ISRCTN 13204258. FINDINGS: The primary outcome was recorded for all participants. We detected no difference between the groups in the need for manual removal of placenta (oxytocin 179/292 [61.3%] vs placebo 177/285 [62.1%]; relative risk 0.98, 95% CI 0.87-1.12; p=0.84). The need for manual removal was higher in the UK (overall 250/361 [69%]) than in Uganda (90/190 [47%]) or Pakistan (16/26 [62%]). Adverse events did not differ between the two groups. INTERPRETATION: Umbilical oxytocin has no clinically significant effect on the need for manual removal for women with retained placenta. FUNDING: WHO, WellBeing of Women, Pakistan Higher Education Commission. PMID- 20004014 TI - Design, syntheses and evaluation of hemocompatible pegylated-antimicrobial polymers with well-controlled molecular structures. AB - In this paper, we have designed and synthesized well-defined pegylated-polymers with tertiary amines from readily available commodity monomers 2 (dimethylamino)ethyl methacrylate (DMAEMA) and oligo(ethylene glycol) methyl ether methacrylate (OEGMA, M(n) approximately 475 Da) by reversible addition fragmentation chain transfer (RAFT) polymerisation. By employing a simple and efficient post-polymerisation functionalization strategy, tertiary amines were quaternized to result in cationic polymers. By the careful selection of the functional halide, X-(CH(2))q-R, (where in X=halide; R=the chemical functionality; q=the number of alkyl spacer between the quaternary ammonium group and R), a series of polymers with well-controlled molecular weight, different amphiphilic balance and chemical functionalities (such as alkyl, primary alcohol, primary amine and carboxylic acid) were readily synthesized. The antimicrobial activities of these cationic polymers were determined against Gram-positive bacteria Bacillus subtilis. Minimum inhibitory concentration (MIC), the polymer concentration to completely inhibit the bacterial growth, was found to be dependent both on the nature of functional group and the hydrophobicity of the polymer. Amongst the functional groups, both the alkyl and the alcohol groups were found to be effective, with MIC values in the range of 20-80 mg/L. The haemolytic properties of polymers were analyzed against mouse red blood cells. The polymers with a short alkyl or hydroxyl group demonstrated little haemolysis, yet retained strong antimicrobial activity. The overall hydrophobicity of the polymer influenced its haemolytic behavior. These polymers can be promising antimicrobial agents. In addition, the approach proposed in this study to atom efficient design and synthesis of antimicrobial polymers from the commercially available monomers can also be applied to develop well-defined functional cationic polymers for various biomedical applications. PMID- 20004015 TI - The roles of Wnt signaling modulators Dickkopf-1 (Dkk1) and Dickkopf-2 (Dkk2) and cell maturation state in osteogenesis on microstructured titanium surfaces. AB - Osteoblast differentiation on tissue culture polystyrene (TCPS) requires Wnt/beta catenin signaling, regulating modulators of the Wnt pathway like Dickkopf-1 (Dkk1) and Dkk2. Osteoblast differentiation is increased on microstructured titanium (Ti) surfaces compared to TCPS; therefore, we hypothesized that surface topography and hydrophilicity affect Dkk1 and Dkk2 expression and that their roles in osteoblast differentiation on Ti differs depending on cell maturation state. Human osteoblast-like MG63 cells, normal human osteoblasts (HOBs), and human mesenchymal stem cells (MSCs), as well as MG63 cells stably silenced for Dkk1 or Dkk2 were grown for 6 days on TCPS and Ti surfaces (PT [Ra<0.2 microm], SLA [Ra=4 microm], modSLA [hydrophilic-SLA]). Dkk1 and Dkk2 mRNA and protein increased on SLA and modSLA for all cell types, but exogenous rhDkk1 and rhDkk2 affected MSCs differently than MG63 cells and HOBs. Silencing Dkk1 reduced MG63 cell number on TCPS and PT, but increased differentiation on these substrates. Silencing Dkk2 reduced stimulatory effects of SLA and modSLA on osteoblast differentiation; Dkk2 but not Dkk1 restored these effects. Antibodies to Dkk1 or Dkk2 specifically blocked substrate-dependent changes caused by the proteins, demonstrating their autocrine action. This indicates major roles for Dkk1 and the canonical Wnt pathway in early-stage differentiation, and for Dkk2 and Wnt/Ca2+ dependent signaling in late-stage differentiation on microstructured and hydrophilic surfaces, during osseointegration. PMID- 20004016 TI - Cell-penetrating macromolecules: direct penetration of amphipathic phospholipid polymers across plasma membrane of living cells. AB - Nanoscaled materials are normally engulfed in endosomes by energy-dependent endocytosis and fail to access the cytosolic cell machinery. Although some biomolecules may penetrate non-endocytically or fuse with plasma membranes without overt membrane disruption, to date no synthetic macromolecule of comparable size has been shown to exhibit this property. Here, we discovered mechanism of direct cell membrane penetration using synthetic phospholipid polymers. These water-soluble amphiphilic phospholipid polymers enter the cytoplasm of living mammalian cells in vitro within a few minutes without overt bilayer disruption even under conditions where energy-dependent endocytic uptakes are blocked. Furthermore, targeted cytosolic distribution to cell organelles was achieved by selecting specific fluorescent tags to the polymers. Thus, the phospholipid polymers can provide a new way of thinking about access to the cellular interior, namely direct membrane penetration. PMID- 20004017 TI - Bioconjugation of barium titanate nanocrystals with immunoglobulin G antibody for second harmonic radiation imaging probes. AB - The second harmonic generation (SHG) active nanocrystals have been demonstrated as attractive imaging probes in nonlinear microscopy due to their coherent, non bleaching and non-blinking signals with a broad flexibility in the choice of excitation wavelength. For the use of these nanocrystals as biomarkers, it is essential to prepare a chemical interface for specific labeling. We developed a specific labeling scheme for barium titanate (BaTiO3) nanocrystals which we use as second harmonic radiation imaging probes. The specificity was achieved by covalently coupling antibodies onto the nanocrystals. We demonstrate highly specific labeling of the nanocrystal conjugates in an antibody microarray and also the membrane proteins of live biological cells in vitro. The development of surface functionalization and bioconjugation of SHG active nanocrystals provides the opportunities of applying them to biological studies. PMID- 20004018 TI - Trophically and topographically functionalized silk fibroin nerve conduits for guided peripheral nerve regeneration. AB - Artificial nerve conduits (NC) can be used as an alternative to autologous nerve grafts to enhance the repair of small nerve gaps. Current NC lack adequate molecular and structural functionalities. Thus, we developed silk fibroin NC (SF NC) that were loaded with glial cell line-derived neurotrophic factor (GDNF) and nerve growth factor (NGF) and topographically functionalized with aligned and non aligned SF nanofibers. The SF NC were produced from fully functionalized SF membranes on which initial experiments were performed. DRG (dorsal root ganglions) sensory neurons and spinal cord (SpC) motor neurons, both from chicken embryos, exhibited an augmented length and rate of axonal outgrowth parallel to the aligned nanofibers. In addition, glial cells from DRG proliferated and migrated in close association and even slightly ahead of the outgrowing axons. On the contrary, axonal and glial growth was slower and randomly oriented on non aligned nanofibers. The DRG and SpC explants were also inserted into the lumen of the finished SF NC. The unidirectional orientation of axo-glial outgrowth from the explants evidenced the preservation of the trophic and topographical functionalities in the SF NC. Bioactive GDNF and NGF were released in vitro from SF NC over 4 weeks. Thus, the developed functionalized SF NC hold promise to enhance functional recovery of injured peripheral nerves. PMID- 20004019 TI - Acute phase response in porcine reproductive and respiratory syndrome virus infection. AB - This study was focused on the changes observed in the serum concentration of haptoglobin (Hp), C-reactive protein (CRP), serum amyloid A (SAA) and Pig-major acute protein (Pig-MAP), during experimental porcine reproductive and respiratory syndrome virus (PRRSV) infection and in their relationship with the expression of interleukin 1beta (IL-1beta), interleukin 6 (IL-6) and tumor necrosis factor alpha (TNF-alpha). Hp and Pig-MAP serum levels were increased at 10 dpi, but CRP and SAA showed a delayed and highly variable increase. All three proinflammatory cytokines were poorly expressed, and only a mild increase in IL-1beta was observed at 7 dpi. The increased expression of Hp coincided with the light enhancement observed in both IL-6 and TNF-alpha, and might be related with an increased expression of IL-10. The low expression of TNF-alpha might point to a possible mechanism of viral evasion of host-immune response. This issue and the delayed expression of CRP and SAA should be taken into account in future studies about modulation of the immune response by PRRSV infection. PMID- 20004020 TI - Structural and functional comparison of MIF ortholog from Plasmodium yoelii with MIF from its rodent host. AB - Host-derived macrophage migration inhibitory factor (MIF) has been implicated in the pathogenesis of malaria infection, especially in malarial anemia. Although two Plasmodium parasite-derived MIF orthologs, Plasmodium falciparum MIF and P. berghei MIF were identified recently, the crystal structure and the precise roles of Plasmodium-derived MIFs, particularly in combination with the host MIF, remain unknown. In this study, we identified another MIF ortholog from a rodent-specific P. yoelii (PyMIF). This molecule shares a conserved three-dimensional structure with murine MIF (MmMIF), but with a different substrate binding pattern and much lower tautomerase activity. It could activate host cells via several signaling pathways in vitro, and inhibiting macrophage apoptosis, also similarly to MmMIF. However, we found that PyMIF and MmMIF acted synergistically to activate the MAPK ERK1/2 signaling pathway at very low concentration but acted antagonistically at higher concentration. Furthermore, we detected PyMIF in the sera of infected mice and found that injection of recombinant PyMIF (rPyMIF) during infection could up regulate several pro-inflammatory cytokines in vivo and slightly delay the death of infected mice. These data suggest that PyMIF modulates host immune responses together with host MIF and has potential to prolong parasitemia or the chronicity of malaria infection. PMID- 20004021 TI - A five-amino-acid motif in the undefined region of the TLR8 ectodomain is required for species-specific ligand recognition. AB - Toll-like receptors play important roles in regulating immunity against microbial infections. Toll-like receptor 8 (TLR8) belongs to a subfamily comprising TLR7, TLR8 and TLR9. Human TLR8 mediates anti-viral immunity by recognizing ssRNA viruses, and triggers potent anti-viral and antitumor immune responses upon ligation by synthetic small molecular weight ligands. Interestingly, distinct from human TLR8, mouse TLR8 was not responsive to ligand stimulation in the absence of polyT-oligodeoxynucleotides (polyT-ODN). The molecular basis for this distinct ligand recognition is still unclear. In the present study, we compared the activation of TLR8 from different species including mouse, rat, human, bovine, porcine, horse, sheep, and cat by ligand ligations. Only the TLR8s from the rodent species (i.e., mouse and rat TLR8s) failed to respond to ligand stimulation in the absence of polyT-ODN. Multiple sequence alignment analysis suggested that these two rodent TLR8s lack a five-amino-acid motif that is conserved in the non-rodent species with varied sequence. This small motif is located in an undefined region of the hTLR8 ectodomain, immediately following LRR 14. Deletion mutation analysis suggested that this motif is essential for the species-specific ligand recognition of hTLR8, whereas it is not required for self dimerization and intracellular localization of this receptor. PMID- 20004022 TI - Maternal sensitivity throughout infancy: continuity and relation to attachment security. AB - Relations among different measures of maternal sensitivity were assessed longitudinally by examining maternal behavior when infants were 4 months, 15 months, and 2.5 years. At each time period, two measures of maternal sensitivity were scored (4 months: maternal vocal and smiling contingency in face-to-face interactions; 15 months: maternal scaffolding and following of infants' play within joint attention; 2.5 years: maternal facilitative and collaborative play within joint attention). When infants were 2.5 years, attachment security was assessed. Mothers' scores on each maternal sensitivity measure within each time period were correlated with their maternal sensitivity scores in another time period, suggesting individual measures of maternal sensitivity were tapping similar aspects across the time periods. Maternal vocal contingency at 4 months was the strongest predictor of infants' attachment security over 2 years later. PMID- 20004023 TI - Impaired flexible decision-making in Major Depressive Disorder. AB - BACKGROUND: Depression is associated with dysfunctional affective states, neuropsychological impairment and altered sensitivity to reward and punishment. These impairments can influence complex decision-making in changing environments. METHODS: The contingency shifting variant Iowa Gambling Task (IGT) was used to assess flexible decision-making performance in a group of medicated unipolar Major Depressive Disorder (MDD) patients (n=19) and a group of healthy control volunteers (n=20). The task comprised the standard IGT followed by a contingency shift phase where decks progressively changed reward and punishment schedule. RESULTS: Patients with MDD showed impaired performance compared to controls during both the standard and the contingency-shift phases of the IGT. Analysis of the contingency-shift phase demonstrated that individuals with depression had difficulties perceiving when a previously bad contingency became good. LIMITATIONS: The present findings have several limitations including small sample size, the possible confounding role of medication and absence of other neuropsychological tests (i.e., executive function). CONCLUSION: Depressed patients show impaired decision-making behaviour in static and dynamic environments. Altered sensitivity to reward and punishment is proposed as the mechanism responsible for the lack of advantageous choices and poor adjustment to a changing environment. PMID- 20004024 TI - Anti-beta2 glycoprotein-I antibody increases the risk of pregnancy-induced hypertension: a case-controlled study. AB - The aim of this study was to evaluate whether anti-beta2 glycoprotein-I antibody (anti-beta2GPI) of the IgG or IgM classes is associated with the development of pregnancy-induced hypertension (PIH) or preeclampsia in the Japanese population. In a case-controlled cohort study, peripheral blood was obtained at 8-14 weeks of gestation from a consecutive series of 1155 women. The case group comprised 36 patients who later developed PIH during the pregnancy. Of the 36 PIH patients, 13 had severe PIH, 18 had preeclampsia and 11 had severe preeclampsia. One hundred and eleven age- and parity-matched women whose pregnancies ended in normal delivery without obstetric complications were selected as controls. We found that a titer of anti-beta2GPI IgG>or=1.0 U/ml was a risk factor for severe PIH (P=0.023, OR 5.7 95%CI 1.4-22.8). In addition, titers of anti-beta2GPI IgM>or=1.2 U/ml was found to be a risk factor for PIH (P=0.001, OR 8.8 95%CI 1.6-47.5). In women positive for anti-beta2GPI but negative for lupus anticoagulant, anti cardiolipin, phosphatidylserine-dependent anti-prothrombin, or kininogen dependent anti-phosphatidylethanolamine antibodies, the presence of anti-beta2GPI was not a significant risk factor for development of PIH or preeclampsia. In conclusion, the presence of anti-beta2GPI antibody represents a risk factor for developing PIH and severe PIH. This finding supports the utility of anti-beta2GPI determination as one of the laboratory criteria for anti-phospholipid syndrome classification. The usefulness of anti-beta2GPI measurement among women without other anti-phospholipid antibodies requires further study. PMID- 20004025 TI - Inhibition of 19S proteasomal regulatory complex subunit PSMD8 increases polyspermy during porcine fertilization in vitro. AB - The 26S proteoasome is a multi-subunit protease specific to ubiquitinated substrate proteins. It is composed of a 20S proteasomal core with substrate degradation activity, and a 19S regulatory complex that acts in substrate recognition, deubiquitination, priming and transport to the 20S core. Inhibition of proteolytic activities associated with the sperm acrosome-borne 20S core prevents fertilization in mammals, ascidians and echinoderms. Less is known about the function of the proteasomal 19S complex during fertilization. The present study examined the role of PSMD8, an essential non-ATPase subunit of the 19S complex, in sperm-ZP penetration during porcine fertilization in vitro (IVF). Immunofluorescence localized PSMD8 to the outer acrosomal membrane, acrosomal matrix and the inner acrosomal membrane. Colloidal gold transmission electron microscopy detected PSMD8 on the surface of vesicles in the acrosomal shroud, formed as a result of zona pellucida-induced acrosomal exocytosis. Contrary to the inhibition of fertilization by blocking of the 20S core activities, fertilization and polyspermy rates were increased by adding anti-PSMD8 antibody to fertilization medium. This observation is consistent with a possible role of PSMD8 in substrate deubiquitination, a process which when blocked, may actually accelerate substrate proteolysis by the 26S proteasome. Subunit PSMD8 co immunoprecipitated with acrosomal surface-associated spermadhesin AQN1. This association indicates that the sperm acrosome-borne proteasomes become exposed onto the sperm surface following the acrosomal exocytosis. Since immunological blocking of subunit PSMD8 increases the rate of polyspermy during porcine fertilization, the activity of the 19S complex may be a rate-limiting factor contributing to anti-polyspermy defense during porcine fertilization. PMID- 20004026 TI - Cognitive impairment following high fat diet consumption is associated with brain inflammation. AB - C57Bl/6 mice were administered a high fat, Western diet (WD, 41% fat) or a very high fat lard diet (HFL, 60% fat), and evaluated for cognitive ability using the Stone T-maze and for biochemical markers of brain inflammation. WD consumption resulted in significantly increased body weight and astrocyte reactivity, but not impaired cognition, microglial reactivity, or heightened cytokine levels. HFL increased body weight, and impaired cognition, increased brain inflammation, and decreased BDNF. Collectively, these data suggest that while different diet formulations can increase body weight, the ability of high fat diets to disrupt cognition is linked to brain inflammation. PMID- 20004027 TI - The combined endoscopic management of congenital laryngeal web. AB - Laryngeal web in the anterior commissure is a rare congenital anomaly often leading to severe dyspnea. Endoscopic procedures based on a simple transsection in these cases may worsen the condition because vocal folds have a tendency for fibrosis and granulation tissue formation after surgical interventions. Thus the traditional treatment of choice is the demanding, externally performed laryngotracheal reconstruction generally with a rib cartilage graft and longer period of stenting. This report presents the successful endoscopic management of a congenital laryngeal web in a 2-year-old boy, who previously underwent an uneffective scar laser transsection that led to excessive glotto-subglottic refibrosis. After the CO(2)-laser transsection the authors applied Mitomycin-C and inserted a combined silicon stent by extra-endolaryngeal technique. After the removal of the stent the patient could be decannulated and his voice improved. The application of these minimally invasive endoscopic techniques was successful, hence it may be an effective alternative treatment option for laryngeal webs. PMID- 20004028 TI - Recent advances and method development for drug target identification. AB - Although it is commonly recognized that most drugs cause inhibition or activation of function by physically binding to one or more gene products, the direct interactions of bioactive small molecules with specific gene products, or targets, is often not well characterized. From a therapeutic perspective, it is nevertheless essential to know a drug's binding partner(s) to understand the mechanism of action and anticipate possible side effects to avoid costly clinical failures. This knowledge is increasingly important as the prevalence of polypharmacy expands to include drugs that engage multiple targets. This review provides a succinct overview of several recent approaches that employ genetics, proteomics, expression profiling or bioinformatics procedures for the systematic characterization of the targets of bioactive compounds. The continuous improvement and advancement of existing technologies is critically discussed and we offer a perspective on the future of innovative emerging new generation technologies. PMID- 20004029 TI - Aging affects the mental simulation/planning of the "rising from the floor" sequence. AB - We investigated the effect of aging on the ability to mentally simulate/plan a complex sequential action of the whole body, namely "rising from the floor". Forty-four non-demented elderly people (mean age: 85.2+/-5.5 years) and 20 young people (mean age: 26.6+/-4.9 years) were included in the study. They were required to put in order six images representing the main movements necessary to get up from a sitting position on the floor. We showed that older subjects had poorer performance-both in terms of proportion of success and response time-than their younger counterparts. These results are in line with previous findings showing age-related alterations in action simulation/action planning processes, and highlight the fact that elderly people have particular difficulties when the action to mentally simulate is complex. PMID- 20004030 TI - Resting heart rate: a predictor of allograft vasculopathy? PMID- 20004031 TI - Percutaneous coronary interventions in Chinese mainland 2008. AB - Percutaneous coronary intervention (PCI) developed rapidly in Chinese mainland in recent years. We collected data on PCIs performed in Chinese mainland in 2008 and analyze the causes of the differences among provinces in PCI development. 182,312 PCIs were performed in 1061 hospitals in 2008. The population-adjusted PCI rate was 137.28 procedures per million inhabitants. PCIs in different provinces had positive correlation with local per capita gross domestic product (GDP) (B=0.06, P<0.05) and with the hospitals which had completed over 100 procedures per year (B=434.23, P<0.05), and had negative correlation with local population (B=0.49, P<0.05). PMID- 20004032 TI - An economic decision support tool for simulating paratuberculosis control strategies in a UK suckler beef herd. AB - A dynamic, deterministic, economic simulation model was developed to estimate the costs and benefits of controlling Mycobacterium avium subsp. paratuberculosis (Johne's disease) in a suckler beef herd. The model is intended as a demonstration tool for veterinarians to use with farmers. The model design process involved user consultation and participation and the model is freely accessible on a dedicated website. The 'user-friendly' model interface allows the input of key assumptions and farm specific parameters enabling model simulations to be tailored to individual farm circumstances. The model simulates the effect of Johne's disease and various measures for its control in terms of herd prevalence and the shedding states of animals within the herd, the financial costs of the disease and of any control measures and the likely benefits of control of Johne's disease for the beef suckler herd over a 10-year period. The model thus helps to make more transparent the 'hidden costs' of Johne's in a herd and the likely benefits to be gained from controlling the disease. The control strategies considered within the model are 'no control', 'testing and culling of diagnosed animals', 'improving management measures' or a dual strategy of 'testing and culling in association with improving management measures'. An example 'run' of the model shows that the strategy 'improving management measures', which reduces infection routes during the early stages, results in a marked fall in herd prevalence and total costs. Testing and culling does little to reduce prevalence and does not reduce total costs over the 10-year period. PMID- 20004033 TI - The additive utility assumption of the QALY model revisited. AB - Quality-adjusted life years are valid representations of the preferences of individuals for health outcomes only under a set of restrictive assumptions. One of the key assumptions is additive utility independence (AUI). Recently, Bleichrodt and Filko [Bleichrodt and Filko, 2008. Journal of Health Economics 27 (5), 1237-1249] presented a new test for AUI, the test for generalized marginality (GM). Based on a student survey showing that violations observed at an individual level cancel out at the group level they concluded that use of the QALY model for economic evaluations in health care is supported. In this comment we argue that this conclusion is not warranted for 2 independent reasons: (i) the GM test is not sufficient to claim AUI both at an individual and group (i.e., aggregated) level and (ii) the student survey is not appropriate to make generalized statements about preferences at the population level. PMID- 20004034 TI - Evaluation of bactericidal activity of weakly acidic electrolyzed water (WAEW) against Vibrio vulnificus and Vibrio parahaemolyticus. AB - Vibrio parahaemolyticus and Vibriovulnificus cause severe foodborne illness in humans; thus, to reduce outbreaks of disease, it is clearly important to reduce food contamination by these pathogens. Although electrolyzed oxidizing (EO) water has been reported to exhibit strong bactericidal activities against many pathogens, it has never been tested against V. vulnificus and V. parahaemolyticus. The purpose of this study was to evaluate the bactericidal activity of weakly acidic electrolyzed water (WAEW), a type of EO water, against V. vulnificus and V. parahaemolyticus. Cell suspensions and cell cultures of both pathogens were treated for 30s with sodium hypochlorite solution containing 35mg/L available chlorine concentration (ACC) or WAEW containing 35mg/L ACC. After an initial inoculum of 5.7logCFU/mL, the number of viable V. vulnificus cells was reduced by 2.2 logs after treatment for 60s with sodium hypochlorite solution containing 35mg/L ACC, while no cells survived treatment with WAEW for 30s. Similar results were obtained for V. parahaemolyticus. Under open storage conditions, WAEW maintained bactericidal activities against cell suspensions of both strains after 5weeks but disappeared against cell cultures of the two strains after 5weeks. Under closed storage conditions, however, WAEW maintained bactericidal activities against both cell suspensions and cell cultures of each strain after 5weeks. No cells were detected in the cell suspensions and cultures when the ACC of WAEW was more than 20mg/L and treatment time was greater than 15s. Bactericidal activity of WAEW against V. vulnificus cell culture was reduced when the ACC of WAEW was less than 15mg/L but was maintained in the V. vulnificus cell suspension when the ACC of WAEW was 0.5mg/L. Thus, the bactericidal activity of WAEW was primarily affected by ACC rather than treatment time. Similar results were obtained for V. parahaemolyticus, indicating that WAEW kills these microorganisms more quickly than a chemical product such as sodium hypochlorite (NaClO), even at equivalent ACCs. PMID- 20004035 TI - Impact of metabolic syndrome, diabetes and prediabetes on cardiovascular events: Tehran lipid and glucose study. AB - AIMS: To compare the cardiovascular disease (CVD) risk associated with the metabolic syndrome (MetS) and dysglycemia, independent of each other during a median follow-up of 6.7 years. METHODS: Study population included 4018 Iranian subjects >or=40 years, free of CVD or coronary heart disease (CHD) at baseline. Incidence rates and hazard ratio (HR) were estimated by the presence or absence of MetS and dysglycemia. Considering the glycemic status, the ability of MetS in prediction of CVD after adjustment for age, sex and CVD risk factors was assessed. RESULTS: The prevalence of MetS, impaired fasting glucose or impaired glucose tolerance (IFG/IGT) and diabetes were 51.4%, 27.3%, and 18.7%, respectively. The addition of MetS to diabetes did not change the CVD risk compared to diabetic subjects without MetS (reference group) after adjustment for age and sex [HR: 1.62 (95% CI 0.93-2.81)] and CVD risk factors [HR: 1.49 (95% CI 0.86-2.60)]. However, addition of MetS to IFG/IGT increased the risk of CVD by 2.5 times compared to those with IFG/IGT without MetS (the risk factor adjusted HR: 2.45 (95% CI 1.32-4.55)). CONCLUSIONS: In Iranian population, MetS did not add to diabetes to predict incident CVD however, IFG/IGT was a significant predictor only in the presence of MetS. PMID- 20004036 TI - Hypomagnesaemia is associated with diabetes: Not pre-diabetes, obesity or the metabolic syndrome. AB - AIMS: The mechanism for the association between diabetes and hypomagnesaemia remains uncertain. This study aimed to test whether hypomagnesaemia is present in pre-diabetes, obesity and the metabolic syndrome. METHODS: 1453 adults from randomly selected households from rural Victoria, Australia, attended for biomedical assessment. Serum magnesium concentrations, hypomagnesaemia defined using local laboratory criteria (<0.70mmol/l), and defined by the bottom quintile of serum magnesium concentrations, were compared in different diabetes pre-cursor states including metabolic syndrome using ATP III criteria. RESULTS: The mean serum magnesium was 0.84+/-0.06mmol/l and 25 (1.7%) had a low magnesium. Mean magnesium was lower among those with known diabetes than those with new diabetes, impaired glucose tolerance (IGT), impaired fasting glucose (IFG) and normal subjects (0.79 (0.78-0.81) vs 0.83 (0.81-0.86); 0.84 (0.82-0.85); 0.84 (0.82 0.86); 0.85 (0.84-0.85)mmol/l). After adjusting for confounders, and compared with those without diabetes, hypomagnesaemia was 10.51 (1.37-80.60)-fold more common with new diabetes, 8.63 (2.20-33.90)-fold more common with known diabetes, 6.77 (1.75-26.17)-fold more common among those taking anti-hypertensive medication but with no difference to those with IGT/IFG (0.90 (0.10-8.10)). CONCLUSION: Diabetes is associated with hypomagnesaemia, but not its pre-cursor states. PMID- 20004037 TI - The Kathmandu Declaration: "Life Circle" approach to prevention and care of diabetes mellitus. AB - OBJECTIVES: To formulate strategies and action plans for the prevention and care of diabetes mellitus as part of the implementation of the International Diabetes Federation (IDF) United Nations Resolution (UNR) 61/225 through a unique concept of a "Life Circle" approach. METHOD: Consensus following review of evidence available and presented at a meeting convened to achieve the objective co-chaired by the IDF President and President elect and diabetologists from several countries in the IDF regions. CONCLUSIONS: The Kathmandu Declaration presents the concept of a "Life Circle" approach to prevention and care of diabetes--a continuum beginning from preconception, pregnancy, infancy and childhood to adult life in an integrated manner. Emphasis is on the benefits on entering the circle at any point and formulates guidelines that could be incorporated in any national diabetes prevention and care programme, indicating the interactive role of all known aetiological factors. PMID- 20004038 TI - Rural-urban differences in health-seeking for the treatment of childhood malaria in south-east Nigeria. AB - OBJECTIVES: To identify the differences in health-seeking for childhood malaria treatment, between urban and rural communities in Nigeria, with a view to providing information to policy makers that will be used to improve malaria control. METHODS: Quantitative and qualitative research methods were employed in eliciting information. A pre-tested structured questionnaire was administered to 1200 caretakers of children under 5 years who had malaria 2 weeks prior to the survey period. Focus group discussions were held with mothers and in-depth interviews with health care providers. RESULTS: Health-seeking for malaria, differed significantly between rural and urban mothers. While majority (64.7%) of urban caretakers patronized private/government health facilities, most (62%) of their rural counterparts resorted to self-treatment with drugs bought over-the counter, from patent medicine vendors. Hospitals were geographically more accessibility to urban than rural dwellers. Rural mothers only go to hospital when the problem persists or becomes worse, which results in delay in seeking appropriate and timely care. CONCLUSION: Urban and rural mothers differed in their responses to childhood fevers. Training drug vendors and caretakers are important measures to improve malaria control. Health facilities with good quality services and readily available drugs should be provided. PMID- 20004039 TI - An incremental knowledge acquisition-based system for supporting decisions in biomedical domains. AB - In Intensive Care Units doctors have to manage several alarm situations in patients. When a doctor analyzes the state of the patient, (s)he has to decide if there is an alarm situation and make decisions about what actions to perform. It is desirable to detect these situations before they occur, because the solution could be easier and the doctor has more time to react. An intelligent system could analyze the information, extract conclusions, format and order the causes leading to the severe condition. This would be helpful for a doctor, and would make the decision-making process easier. A system capable of performing such operations is presented here. This is not a diagnosis application but a tool to detect alarm situations for patient safety. A prototype capable of making retrospective evaluation of the condition of the patients has been developed. This system is based on the MCRDR technology, which has been extended to deal with the requirements of this domain. The evaluation of the system is also reported in this paper. PMID- 20004040 TI - Are adenosquamous lung carcinomas a simple mix of adenocarcinomas and squamous cell carcinomas, or more complex at the molecular level? AB - Adenocarcinomas (AC), squamous cell carcinomas (SCC) and adenosquamous carcinomas (ASC) are three histological subtypes of non-small-cell lung carcinomas (NSCLC). ASC are morphologically mixed tumours that contain the two cell components AC and SCC. To understand if they are a "simple" mix of AC and SCC or if they present molecular specificities, as compared with the molecular characterization of both components, we performed a comparative transcriptome analysis on a series of nine ASC, five AC and five SCC induced in rats by radon exposure. We found that 72, 40 and 39 genes were differentially expressed when comparing AC_SCC, ASC_SCC and AC_ASC, respectively. Moreover, when classifying the three histological subtypes, using genes that discriminated AC and SCC, we observed that all ASC were classified as intermediate between the AC and SCC, some being closer to AC, others to SCC. These results indicated that, regarding gene expression, ASC could be considered as a mix of AC and SCC, both in various proportions. However, they also exhibit molecular specificities since we found specific genes discriminating ASC_SCC and AC_ASC. In conclusion, the ASC mixed lung tumours are more complex than simple mixes of AC and SCC components. Neuroendocrine differentiation and ERK proliferation pathways seemed preferentially deregulated in ASC compared to AC and SCC respectively, pathways that are worthy of being explored because they could partially explain the high clinical aggressiveness of ASC. PMID- 20004041 TI - Follow-up study of the GIGYF2 gene in French families with Parkinson's disease. AB - Meeus et al. (2009) reported no pathogenic mutations in a comprehensive genetic analysis of the entire GRB10-interacting GYF protein-2 gene (GIGYF2) in a Belgian series of both familial and sporadic patients with Parkinson's disease (PD). Although we initially proposed that GIGYF2 corresponds to the PARK11 locus, in familial PD, we found no causative variations in a follow-up study in which GIGYF2 was screened in an independent series of 185 patients with autosomal dominant PD, mostly of French origin. Together, these results do not support a major role of GIGYF2 in PD. PMID- 20004042 TI - [Health policies of national and regional level for the immigrant population in Spain]. AB - OBJECTIVES: To analyze the content of health policies for the immigrant population developed by central and regional governments in Spain. METHODS: A descriptive comparative study of central and regional healthcare policies for the immigrant population was conducted in Spain through content analysis. The selected regions were Andalusia, Valencia, Madrid and the Basque Country as these regions have specific policies, distinct proportions of immigrants and policy evaluations. National or regional health and immigration plans with health policies for immigrants were selected. Contents analysis was conducted of the following main dimensions: policy principles and objectives, strategies and results' evaluation. Subsequently, strategies were categorized according to the area of intervention. RESULTS: Healthcare policies for the immigrant population are mainly included in national and regional immigration plans. The principles of these policies are based on equal rights to healthcare between the immigrant and native-born populations and the objectives aim to achieve this end. National objectives and actions address access to and adaptation of health services, health promotion, health needs assessment, and health personnel training in cultural competences. Regional policies follow the national guidelines but their actions are more specific. Policy evaluations are highly limited. CONCLUSIONS: The content of the health policies, especially national policies, address major issues in meeting immigrants' healthcare needs. However, the absence of assessments, together with persistent problems in the provision of care and inequalities in access, could indicate insufficient implementation and requires careful monitoring. PMID- 20004043 TI - Synthesis, characterization and biological activity of some new 1,3,4-oxadiazole bearing 2-flouro-4-methoxy phenyl moiety. AB - In the present study a series of new 1,3,4-oxadiazole derivatives containing 2 fluoro-4-methoxy moiety were synthesized. These newly synthesized compounds were characterized by NMR, mass spectral, IR spectral study and also by C, H, N analyses. All the newly synthesized compounds were screened for their antibacterial and antifungal studies. Antimicrobial studies revealed that compounds 4a and 4b showed significant antibacterial activity against Escherichia coli and Pseudomonas aeruginosa Compound 4i showed significant antifungal activity against C. albicans. PMID- 20004044 TI - Synthesis and antitumor activity of new dihydropyridine thioglycosides and their corresponding dehydrogenated forms. AB - A number of a new pyridine thioglycosides were synthesized via reaction of piperidinium salts of dihydropyridinethiones with 2,3,4,6-tetra-O-acetyl-alpha-D gluco- and galactopyranosyl bromide. The antitumor activities of the synthesized compounds were evaluated utilizing two different human cell lines. Some of the tested compounds showed high inhibition of human cell lines. The detailed synthesis, spectroscopic data and antitumor activities for the synthesized compounds were reported. PMID- 20004045 TI - Curcuminoid analogs with potent activity against Trypanosoma and Leishmania species. AB - The natural curcuminoids curcumin (1), demethoxycurcumin (2) and bisdemethoxycurcumin (3) have been chemically modified to give 46 analogs and 8 pairs of 1:1 mixture of curcuminoid analogs and these parent curcuminoids and their analogs were assessed against protozoa of the Trypanosoma and Leishmania species. The parent curcuminoids exhibited low antitrypanosomal activity (EC(50) for our drug-sensitive Trypanosoma brucei brucei line (WT) of compounds 1, 2 and 3 are 2.5, 4.6 and 7.7 microM, respectively). Among 43 curcuminoid analogs and 8 pairs of 1:1 mixture of curcuminoid analogs tested, 8 pure analogs and 5 isomeric mixtures of analogs exhibited high antitrypanosomal activity in submicromolar order of magnitude. Among these highly active analogs, 1,7-bis(4-hydroxy-3 methoxyphenyl)hept-4-en-3-one (40) was the most active compound, with an EC(50) value of 0.053+/-0.007 microM; it was about 2-fold more active than the standard veterinary drug diminazene aceturate (EC(50) 0.12+/-0.01 microM). Using a previously characterized diminazene-resistant T. b. brucei (TbAT1-KO) and a derived multi-drug resistant line (B48), no cross-resistance of curcuminoids was observed to the diamidine and melaminophenyl arsenical drugs that are the current treatments. Indeed, curcuminoids carrying a conjugated keto (enone) motif, including 40, were significantly more active against T. b. brucei B48. This enone motif was found to contribute to particularly high trypanocidal activity against all Trypanosoma species and strains tested. The parent curcuminoids showed low antileishmanial activity (EC(50) values of compounds 1 and 2 for Leishmania mexicana amastigotes are 16+/-3 and 37+/-6 microM, respectively) while the control drug, pentamidine, displayed an EC(50) of 16+/-2 microM. Among the active curcuminoid analogs, four compounds exhibited EC(50) values of less than 5 microM against Leishmania major promastigotes and four against L. mexicana amastigotes. No significant difference in sensitivity to curcuminoids between L. major promastigotes and L. mexicana amastigotes was observed. The parent curcuminoids and most of their analogs were also tested for their toxicity against human embryonic kidney (HEK) cells. All the curcuminoids exhibited lower toxicity to HEK cells than to T. b. brucei bloodstream forms and only one of the tested compounds showed significantly higher activity against HEK cells than curcumin (1). The selectivity index for T. b. brucei ranged from 3-fold to 1500-fold. The selectivity index for the most active analog, the enone 40, was 453-fold. PMID- 20004046 TI - A theoretical study of the structure-radical scavenging activity of trans resveratrol analogues and cis-resveratrol in gas phase and water environment. AB - Quantum calculations based on the density functional theory (DFT) have been employed to study the relation between the structure and antioxidant activity of trans-resveratrol (TR), cis-resveratrol (CR), trans-4.4'-dihydroxystilbene (trans 4,4'-DHS), trans-3,4-dihydroxystilbene (trans-3,4-DHS), trans-3,4,4' trihydroxystilbene (trans-3,4,4'-THS), trans-3,4,5-trihydroxystilbene (trans 3,4,5-THS) and alpha,beta-dihydro-3,4',5-trihydroxystilbene (alpha,beta-dihydro 3,4',5-THS) in the gas phase and water environment. We have shown that the antioxidant activity of trans-stereoisomers of the compounds considered is related to their phenoxy radicals showing a planar and semiquinone conformation that allows delocalization of the unpaired electron through the whole trans stilbene skeleton. The calculations have revealed that trans-3,4-DHS, trans 3,4,4'-THS, trans-3,4,5-THS and trans-4,4'-DHS exhibit higher antioxidant activity than TR, while alpha,beta-dihydro-3,4',5-THS and CR are less efficient antioxidants than TR. We have proved that all compounds have higher ability to donate hydrogen atom in the gas phase than in the presence of water medium in which they are very sensitive to electron donation. The results obtained demonstrate that the H-transfer mechanism for scavenging of the free radicals is more preferable than the single-electron transfer in both types of environment. They are consistent with the experiments confirming the specific biological activity of the compounds considered. PMID- 20004047 TI - Fractionation of plutonium in environmental and bio-shielding concrete samples using dynamic sequential extraction. AB - Fractionation of plutonium isotopes ((238)Pu, (239,240)Pu) in environmental samples (i.e. soil and sediment) and bio-shielding concrete from decommissioning of nuclear reactor were carried out by dynamic sequential extraction using an on line sequential injection (SI) system combined with a specially designed extraction column. Plutonium in the fractions from the sequential extraction was separated by ion exchange chromatography and measured using alpha spectrometry. The analytical results show a higher mobility of plutonium in bio-shielding concrete, which means attention should be paid to the treatment and disposal of nuclear waste from decommissioning. PMID- 20004048 TI - Re: one-stage management of ankylosis of the temporomandibular joint with a custom-made total joint replacement system. PMID- 20004049 TI - ASA grade and disease-free mortality in head and neck cancer patients: a prospective study. AB - Complex surgery with curative intent as part of the care of patients with head and neck cancer, who also have serious coexisting conditions is sometimes viewed critically as being unduly, optimistic. We have used American Society of Anesthesiologists' (ASA) grading by a single anaesthetist prospectively as a baseline to investigate a possible link between coexisiting conditions and disease-free survival in 114 patients with head and neck cancer patients treated by the same anaesthetist and surgical team, and found that the ASA grade is not a reliable predictor of disease-free survival. There was no significant association between ASA grade and overall mortality, but there was a significant association between ASA grade and mortality associated with metastatic disease. However, the test for trend was not significant, which suggested that deaths from metastatic disease did not increase in line with ASA grading. All patients in ASA grades II and III were alive 2 years after their initial operation and the risk of mortality after 2 years may increase by up to 10%. PMID- 20004050 TI - Assessment of nematode community structure as a bioindicator in river monitoring. AB - Nematode communities from river water and sediments were assessed for the abundance, feeding types, maturity indices and nematode channel ratio (NCR). The sampling sites studied included different levels of pollution and contamination from agricultural, industrial and sewage sources. The nematode abundance found in the sediment samples was more than that in the water samples. The lowest nematode abundance in sediment samples and the lowest NCR in water samples were both found at the industrial pollution site. Water samples showed positive correlation between the NCR and river pollution index (RPI). Mean maturity indices in sediment samples were inversely correlated with RPI. The pollutant source determined the relationship between NCR and pollution level, while maturity index always showed negative correlation with pollutant level regardless of the pollutant sources. The nematode abundance and its community structure were both reliable bioindicators for monitoring long-term river pollution in both qualitative and quantitative aspects. PMID- 20004051 TI - Diagnosis of COPD in hospitalised patients. AB - OBJECTIVE: To examine the quality of COPD diagnosis in hospitalised patients. MATERIAL AND METHODS: Retrospective multicentre cross-sectional audit review of the clinical histories of patients discharged with a diagnosis of COPD. The diagnosis of COPD was considered correct (DxC) in cases where the combination of a bronchial obstruction (FEV1/FVC<70%) and smoking (>10 packets/year) could be documented. In the rest of the cases the diagnosis was considered deficient (DxD). A DxC in at least 60% of patients was required to be considered an acceptable quality health care diagnosis. Demographic data such as, smoking, spirometry, the specialist who discharged the patient (P: Pneumologist; MS: Medical Specialty; CS: Surgical Specialty), and health care level (hospital complexity; low (H1), intermediate (H2) and high (H3)). RESULTS: A total of 840 cases were analysed (718 males, 122 females); mean age (SD) 73 (10), from 10 hospitals (3 H1, 4 H2, 3 H3). A DxD was obtained in 597 (71.1%), due to either lack of spirometry (538, 64%) or smoking criteria (319, 38%), (P<0.001). Only two of the ten hospitals complied with the criteria of an acceptable quality health care diagnosis. Significant differences (P<0.0001) were seen on comparing DxC and DxD by health care level (DxC: 56.2% in H1, 29.9% in H2, 20.9% in H3), and by specialist (DxC: 47.6% en P, 24.6% in SP, 17.4% in MS). A multivariate analysis associated DxC with the male sex, H1 and pneumology reports. CONCLUSIONS: 1. The quality health care for the diagnosis of COPD is deficient. 2. The lack of spirometry is the most common cause of DxD. PMID- 20004052 TI - Photodynamic diagnosis in non-muscle-invasive bladder cancer: a systematic review and cumulative analysis of prospective studies. AB - CONTEXT: The clinical benefit of photodynamic diagnosis (PDD) with 5 aminolevulinic acid or hexaminolevulinate in addition to white-light cystoscopy (WLC) in bladder cancer has been discussed controversially. OBJECTIVE: To assess in a systematic review the effect of PDD in addition to WLC on (1) the diagnosis and (2) the therapeutic outcome of primary or recurrent non-muscle-invasive bladder cancer investigated by cystoscopy or transurethral resection. EVIDENCE ACQUISITION: An electronic database search of Medline, Embase, the Cochrane Library, and CancerLit was undertaken, plus hand searching of relevant congress abstracts and urologic journals. Trials were included if they prospectively compared WLC with PDD in bladder cancer. The review process followed the guidelines of the Cochrane Collaboration. Two reviewers evaluated independently both trial eligibility and methodological quality and data extraction. EVIDENCE SYNTHESIS: The primary end point of diagnostic accuracy was additional detection rate. The primary end points of therapeutic outcome were residual tumour at second resection and recurrence-free survival (RFS). Seventeen trials were identified. Twelve diagnostic trials used WLC and PDD with the same patients. Seven reported results for the subgroup of patients with carcinoma in situ (CIS). Five randomised trials studied therapeutic outcome. The results were combined in random effects meta-analyses if end points, designs, and populations were comparable. Twenty percent (95% confidence interval [CI], 8-35) more tumour positive patients were detected with PDD in all patients with non-muscle-invasive tumours and 39% (CI, 23-57) more when only CIS was analysed. Heterogeneity was present among diagnostic studies even when the subgroup of patients with CIS was investigated. Residual tumour was significantly less often found after PDD (odds ratio: 0.28; 95% CI, 0.15-0.52; p<0.0001). RFS was higher at 12 and 24 mo in the PDD groups than in the WLC-only groups. The combined p value of log-rank tests of RFS was statistically significant (0.00002). CONCLUSIONS: PDD detects more bladder tumour-positive patients, especially more with CIS, than WLC. More patients have a complete resection and a longer RFS when diagnosed with PDD. PMID- 20004053 TI - Re: Hans-Martin Fritsche, Maximilian Burger, Robert S. Svatek et al. Characteristics and outcomes of patients with clinical T1 grade 3 urothelial carcinoma treated with radical cystectomy: results from an international cohort. Eur Urol 2010;57:300-9. PMID- 20004054 TI - Prognostication of upper tract urothelial carcinoma: what do we really need? PMID- 20004055 TI - Detection of explosives by positive corona discharge ion mobility spectrometry. AB - In this work, thermal decomposition has been used to detect explosives by IMS in positive polarity. Explosives including Pentaerythritol Tetranitrate (PETN), Cyclo-1,3,5-Trimethylene-2,4,6-Trinitramine (RDX), 2,4,6-Trinitrotoluene (TNT), 2,4-Dihydro-5-nitro-3H-1,2,4-triazol-3-one (NTO), 1,3,5,7-Tetranitro-1,3,5,7 tetrazocine (HMX), have been evaluated at temperatures between 150 and 250 degrees C in positive polarity in air. Explosives yield NO(x) which causes NO(+) peak to increase. Additional peaks may be used to identify the type of explosive. The limit of detection for RDX, HMX, PETN, NTO, and TNT were obtained to be 1, 10, 40, 1000, and 1000 ng, respectively. PMID- 20004056 TI - Cr(Vi) reduction capacity of activated sludge as affected by nitrogen and carbon sources, microbial acclimation and cell multiplication. AB - The objectives of the present work were: (i) to analyze the capacity of activated sludge to reduce hexavalent chromium using different carbon sources as electron donors in batch reactors, (ii) to determine the relationship between biomass growth and the amount of Cr(VI) reduced considering the effect of the nitrogen to carbon source ratio, and (iii) to determine the effect of the Cr(VI) acclimation stage on the performance of the biological chromium reduction assessing the stability of the Cr(VI) reduction capacity of the activated sludge. The highest specific Cr(VI) removal rate (q(Cr)) was attained with cheese whey or lactose as electron donors decreasing in the following order: cheese whey approximately lactose>glucose>citrate>acetate. Batch assays with different nitrogen to carbon source ratio demonstrated that biological Cr(VI) reduction is associated to the cell multiplication phase; as a result, maximum Cr(VI) removal rates occur when there is no substrate limitation. The biomass can be acclimated to the presence of Cr(VI) and generate new cells that maintain the ability to reduce chromate. Therefore, the activated sludge process could be applied to a continuous Cr(VI) removal process. PMID- 20004057 TI - Hybrid signal processing in voltammetric determination of chromium(VI). AB - This study presents different hybrid signal processing algorithms which are useful in the interpretation of voltammetric signals recorded on mercury film electrodes for the determination of Cr(VI). Because of the complex character of the distortions (random, fast increasing, nonlinear, background noise and other perturbations) the application of a complex numerical procedure is necessary. In this work different variants of hybrid algorithms are utilized: adaptive degree polynomial filter, baseline generation and subtraction, signals ratio method, orthogonal signal correction and continuous wavelet transformation with dedicated mother wavelet. The operation and effectiveness of proposed procedures were tested by the determination of very low concentrations of Cr(VI) in synthetic and river water samples. PMID- 20004058 TI - Combination of cloud point extraction and flame atomic absorption spectrometry for preconcentration and determination of nickel and manganese ions in water and food samples. AB - A simple, rapid, inexpensive, and nonpolluting cloud point extraction (CPE) technique has been improved for the preconcentration and determination of nickel and manganese. After complexation with p-nitrophenylazoresorcinol (Magneson I), the analytes could be competitively extracted in a surfactant octylphenoxy polyethoxyethanol (Triton X-114), prior to determination by flame atomic absorption spectrometry (FAAS). The effects of experimental conditions such as pH, concentration of chelating agent and surfactant, equilibration temperature and time on CPE were studied. Under the optimum conditions, preconcentration of a 25 mL sample solution permitted the detection of 2.7 ng mL(-1) Ni(2+) and 2.9 ng mL(-1) Mn(2+) with enrichment factors of 17 and 19 for Ni(2+) and Mn(2+), respectively. The developed method was applied to the determination of trace nickel and manganese in water and food samples with satisfactory results. PMID- 20004059 TI - Operational characteristics of efficient co-removal of H2S and NH3 in a horizontal biotrickling filter using exhausted carbon. AB - Odorous H(2)S and NH(3) gases were effectively biodegraded in a horizontal biotrickling filter (HBTF) packed with H(2)S-exhausted carbon. During the process, the operational characteristics including the distribution of degradation products, biomass accumulation and biological activity, carbon surface characteristics and pressure drop were investigated. The results show that the content of biodegradation products on carbon was low, i.e. 0.9-2.8 wt% S and 0.3-1.0 wt% N. The low content benefited the stable operation of the HBTF, due to preventing a toxic concentration of degradation products on packing bed over a long-term operation. The biomass was distributed evenly along the HBTF . This avoided the problems of bed clogging and activity loss. On the other hand, the deteriorated performance was observed due to the biomass accumulation over a long-term operation. Carbon surface characteristics in the HBTF remained almost unchanged. pH values of carbon were neutral and micropore structure of carbon remained relatively stable. In addition, the pressure drop in the HBTF was very low. These operational characteristics of the HBTF system significantly contributed to efficient co-removal of H(2)S and NH(3) in the HBTF over a long term operation. PMID- 20004060 TI - Central sensitization in carpal tunnel syndrome with extraterritorial spread of sensory symptoms. AB - Extraterritorial spread of sensory symptoms is frequent in carpal tunnel syndrome (CTS). Animal models suggest that this phenomenon may depend on central sensitization. We sought to obtain psychophysical evidence of sensitization in CTS with extraterritorial symptoms spread. We recruited 100 unilateral CTS patients. After selection to rule out concomitant upper-limb causes of pain, 48 patients were included. The hand symptoms distribution was graded with a diagram into median and extramedian pattern. Patients were asked on proximal pain. Quantitative sensory testing (QST) was performed in the territory of injured median nerve and in extramedian territories to document signs of sensitization (hyperalgesia, allodynia, wind-up). Extramedian pattern and proximal pain were found in 33.3% and 37.5% of patients, respectively. The QST profile associated with extramedian pattern includes: (1) thermal and mechanic hyperalgesia in the territory of the injured median nerve and in those of the uninjured ulnar and radial nerves and (2) enhanced wind-up. No signs of sensitization were found in patients with the median distribution and those with proximal symptoms. Different mechanisms may underlie hand extramedian and proximal spread of symptoms, respectively. Extramedian spread of symptoms in the hand may be secondary to spinal sensitization but peripheral and supraspinal mechanisms may contribute. Proximal spread may represent referred pain. Central sensitization may be secondary to abnormal activity in the median nerve afferents or the consequence of a predisposing trait. Our data may explain the persistence of sensory symptoms after median nerve surgical release and the presence of non-anatomical sensory patterns in neuropathic pain. PMID- 20004061 TI - Assessment of amniotic and polyurethane membrane dressings in the treatment of burns. AB - As allograft and xenografts are not available in Islamic countries, amniotic membrane seems to be an effective alternative in the management of deep burns. Its proven bioactivities and modest price suggest that it might be superior to synthetic dressings. Forty-six patients were enrolled in this randomized, controlled clinical trial conducted in the Burn Unit at Suez Canal University Hospital, Ismailia, Egypt. All age groups and both gender were included in the study. Only patients with less than 50% total body surface area burned were included, thus minimizing the dropouts in both groups. All were either second or third degree. These patients were randomly assigned either to group I: amniotic membrane (Biomembrane) dressing, or group II: polyurethane membrane (Tegaderm) dressing. Those in group I demonstrated a significantly lower rate of infection and required less frequent dressing changes than those in group II. They also sustained less electrolyte and albumin loss. The rate of healing in the amniotic membrane group was significantly faster than in the polyurethane group. Furthermore, pain was significantly less when Biomembrane was used. Based on these findings, we recommend the use of lyophilized gamma-irradiated amniotic membrane as an effective alternative for allograft and xenografts in Islamic countries and the Jewish population. PMID- 20004062 TI - Psychometric validation of the Leeds Dependence Questionnaire (LDQ) in a young adult clinical sample. AB - OBJECTIVE: Measures of substance dependence severity that are both clinically efficient and sensitive to change can facilitate assessment of clinical innovation necessary for improving current evidence-based practices. The Leeds Dependence Questionnaire (LDQ) is a 10-item, continuous, self-report measure of dependence that is not specific to any particular substance and has shown promise in preliminary psychometric research. The present study investigates its psychometric properties in a large clinical sample of young adults. METHOD: Young adults (N=300) were enrolled in a naturalistic treatment process and outcome study of residential substance dependence treatment (mean age 20.4 [1.6], range 18-25; 27% female; 95% White). Dependence severity by demographic and diagnostic groupings, factor structure and internal consistency, and criterion- and construct-related validity were examined. RESULTS: Dependence severity in this cohort of youth overall was high (M=18.65 [8.65]). LDQ scores were highest among opiate and stimulant users, and there was a trend for higher scores among women compared to men (t=1.869, p=.063). Factor analysis using a robust alpha factoring extraction revealed a single factor accounting for 63% of the variance in reported dependence severity. The internal consistency was also very high (alpha=.93). Concurrent and convergent validity with dependence criteria, substance use frequency, and general symptom severity, respectively, were also acceptable. CONCLUSIONS: The LDQ shows considerable promise as a brief, psychometrically sound, measure of substance dependence useful across a variety of substances, that has clinical and research utility. This study supports its use among young adults. PMID- 20004063 TI - Beta-lactam antibiotic reduces morphine analgesic tolerance in rats through GLT-1 transporter activation. AB - Glutamate transporter subtype 1 (GLT-1) activation is a promising - and understudied - approach for managing aspects of morphine tolerance caused by increased glutamatergic transmission. Identification of beta-lactam antibiotics as pharmaceuticals which activate GLT-1 transporters prompted us to hypothesize that repeated beta-lactam antibiotic (ceftriaxone) administration blocks development of tolerance to morphine antinociception through GLT-1 activation. Here, we injected rats with morphine (10mg/kg, s.c.) twice daily for 7 days to induce tolerance and used the hot-plate assay to determine antinociception on days 1, 4 and 7 of repeated morphine administration. Ceftriaxone and a selective GLT-1 transporter inhibitor dihydrokainate (DHK) were co-administered with morphine to determine if GLT-1 activation mediated the ceftriaxone effect. Tolerance was present on days 4 and 7 of repeated morphine administration. Ceftriaxone (50, 100 or 200mg/kg, i.p.) administration dose-dependently blocked development of morphine tolerance. DHK (10mg/kg, s.c.), administered 15 min before each morphine injection, prevented inhibition of morphine tolerance by ceftriaxone (200mg/kg, i.p.). These results identify an interaction between ceftriaxone and morphine in opioid-tolerant rats and suggest beta-lactam antibiotics preserve analgesic efficacy during chronic morphine exposure. PMID- 20004064 TI - The intergenerational transmission of tobacco smoking--the role of parents' long term smoking trajectories. AB - Youths whose parents smoke tobacco may be at elevated risk of smoking themselves. However, the association between parental long-term smoking history and offspring regular tobacco use is not well known. Using data collected on 1121 youths (12-26 years) participating in the GAZEL Youth study, a French community-based cohort, we tested the association between parental long-term smoking trajectory and offspring regular smoking. Parental smoking trajectory over 11 years (1989-1999) was measured by yearly reports obtained from the parent. Statistical analyses controlled for youth's sex, age, alcohol use and disruptive behavioral problems, parent's sex, as well as family socioeconomic position. Overall, 27% of study youths smoked regularly. Compared to offspring of non-smokers, those of persistent smokers had twofold smoking rates (age and sex-adjusted OR: 1.91, 95% CI: 1.30-2.79, fully-adjusted OR: 1.96, 95% CI: 1.31-2.93). Additionally, persistent parental smoking predicted offspring heavy smoking and early smoking initiation. Overall, maternal smoking was more strongly associated with youths' regular smoking than paternal smoking (fully-adjusted ORs: 3.12, 95% 1.58-6.16 vs. 1.47, 95% 0.87-2.49). These results suggest that efforts to decrease the burden of tobacco smoking among youths may be more efficient if focused on families rather than on individuals. PMID- 20004065 TI - Nicotine patch vs. nicotine lozenge for smoking cessation: an effectiveness trial coordinated by the Community Clinical Oncology Program. AB - BACKGROUND: Nicotine replacement therapies are efficacious for treating nicotine dependence. However, limited data exist on benefits of different NRTs and predictors of treatment outcome. This study compared the effectiveness of transdermal nicotine vs. nicotine lozenge for smoking cessation and identified predictors of treatment response. METHODS: A randomized, open-label effectiveness trial was conducted at 12 medical sites participating in the National Cancer Institute's Community Clinical Oncology Program. The sample consisted of 642 treatment-seeking smokers randomized to 12 weeks of transdermal nicotine or nicotine lozenge. RESULTS: Smoker characteristics were assessed at baseline, and 24-h point prevalence abstinence confirmed with breath carbon monoxide (CO) was evaluated at end of treatment (EOT) and at a 6-month follow-up. There was a trend for higher quit rates for transdermal nicotine vs. nicotine lozenge at EOT (24.3% vs. 18.7%, p=.10) and 6 months (15.6% vs. 10.9%, p=.10). A logistic regression model of EOT quit rates showed smokers who preferred transdermal nicotine, were not reactive to smoking cues, and did not use nicotine to alleviate distress or stimulate cognitive function had higher quit rates on transdermal nicotine. A logistic regression model of 6-month quit rates showed smokers who preferred transdermal nicotine had higher quit rates on transdermal nicotine, and smokers who used nicotine to alleviate distress or stimulate cognitive processes had lower quit rates on nicotine lozenge. CONCLUSIONS: Transdermal nicotine may be more effective than nicotine lozenge for smokers who prefer transdermal nicotine and do not smoke to alleviate emotional distress or stimulate cognitive function. PMID- 20004066 TI - Treponema pedis isolated from a sow shoulder ulcer. PMID- 20004067 TI - Serodiagnosis of Mycobacterium avium infections in pigs. AB - The aim of this study is the development and evaluation of a serodiagnostic assay for Mycobacterium avium (MA). After screening MA lipid fractions in an ELISA format, a polar lipid fraction was selected as antigen because of its superior recognition by serum antibodies in experimentally infected pigs. The resulting MA ELISA was evaluated as an alternative for detection of MA infection by traditional pathological examination of pig lymph nodes for granulomatous lesions by meat inspectors. By comparing with bacteriological examination, the MA-ELISA showed significantly better sensitivity (69%) as compared to pathological examination (31%) in experimentally infected pigs. The MA-ELISA also appeared significantly more specific in a set of serum samples from MA negative pigs: only 1 out of these 153 serum samples reacted positive, whereas 99 (65%) of these had displayed false positive results by detection of lymph nodes lesions that appeared not to be associated with MA (Komijn et al., 2007). The MA-ELISA was subsequently evaluated using serum samples from two farms with pigs known to be infected with MA. Bacteriological examination of the sub-maxillary and mesenteric lymph nodes showed that 56% (103/184) and 35% (41/117) of the pigs, respectively were positive for MA in these farms. In the first farm, 16% (29/184) of the pigs tested positive in MA-ELISA and 31% (57/184) by pathological examination. On the contrary, in the second farm, more pigs tested positive 17% (15/117) in MA-ELISA with 8% (9/117) positivity by pathological examination. Taking the results on both farms together, the sensitivity of the MA-ELISA was 14% and the specificity 83%, whereas the sensitivity of the pathological examination was 31% and the specificity 86%. For practical reasons use of a serological test as the MA-ELISA may be preferred over pathological or bacteriological examinations. Our studies in experimentally infected and negative "field" sera indicate that the MA-ELISA is significantly more specific and more sensitive than detection by classical pathological examination. However, the studies in two MA infected farms show a variable picture with pathological examination overall performing better. Study in a wider range of "positive" farms will be needed to provide a more comprehensive view of the quality of both tests for detection of MA in infected farms. At the same time further optimization of MA-ELISA with use of lipid antigens from a broader range of serotypes may improve its performance in the face of infections with different MA serotypes. PMID- 20004068 TI - Pharyngeal, rectal and nasal colonization of clinically healthy dogs with Staphylococcus aureus. PMID- 20004069 TI - Exercise training associated with estrogen therapy induced cardiovascular benefits after ovarian hormones deprivation. AB - Menopause is recognized as a period of increased risk for coronary heart disease. Although the benefits of exercise training in lowering cardiovascular risk factors are well established, the risks and benefits of hormone therapy have been questioned. The purpose of the present study was to investigate the effects of estrogen therapy (HT) associated or not with exercise training (ET) in autonomic cardiovascular control in ovariectomized (OVX) rats. Female rats were divided into: control, OVX, OVX+HT, OVX+ET and OVX+HT+ET. HT was performed using a 0.25mg 8-weeks sustained release pellet. Trained groups were submitted to an 8-week exercise training protocol on treadmill. Baroreflex sensitivity (BRS) was evaluated by heart rate responses to arterial pressure (AP) changes, and vagal and sympathetic tonus by pharmacological blockade. Ovariectomy induced an AP increase (123+/-2mmHg vs. 108+/-2mmHg), BRS impairment ( approximately 69%), sympathetic activation ( approximately 100%) and vagal tonus reduction ( approximately 77%) compared to controls. HT or ET normalized the changes in parasympathetic tonus. However, only the association HT+ET was able to promote normalization of AP, BRS and sympathetic tonus, as compared to controls. These results indicate that ET induces cardiovascular and autonomic benefits in OVX rats under HT, suggesting a positive role of this association in the management of cardiovascular risk factor in postmenopausal women. PMID- 20004070 TI - A case of intraneural perineurioma presenting with monomelic atrophy in a child. AB - We report the case of an 11-year-old girl who developed slowly progressive atrophy of the left lower extremity. She suffered from mild dilated cardiomyopathy of unknown cause since 4years of age. When she was 7years old, her family noticed that her left extremity was thinner compared to the right one. Computed tomography showed atrophy and areas of low density in the left gluteus maximus, thigh, and calf muscles. The left sciatic nerve showed gadolinium enhancement on magnetic resonance imaging. A biopsy of the left sural nerve revealed pseudo-onion bulbs. Immunohistochemical staining was positive for epithelial membrane antigen and negative for S100 protein. Electron microscopy demonstrated myelinated or unmyelinated nerve fibers surrounded by concentric layers of perineurial cells. These results indicated intraneural perineurioma. The tumor was estimated at least from the nerve root to the ankle joint. The length of nerve involvement in this patient was the highest recorded in the literatures. Intraneural perineurioma is a very rare disorder, but is tend to be found in youth. This disorder should be considered when we see children with monomelic weakness and/or atrophy. PMID- 20004071 TI - [Anal fistula]. PMID- 20004072 TI - Pre-treatment hemodynamic features involved with long-term survival of cirrhotic patients after embolization of gastric fundal varices. AB - PURPOSE: To clarify the pre-treatment hemodynamic features involved in the long term survival of cirrhotic patients with gastric fundal varices (FV) after balloon-occluded retrograde transvenous obliteration (B-RTO). MATERIALS AND METHODS: Eighty-one cirrhotic patients with medium- or large-grade FV treated by B-RTO were enrolled in this retrospective study. Pre-treatment flow volume ratio between gastric vein and portal trunk (GP-R) was obtained by Doppler ultrasound. RESULTS: The cumulative survival rate was 90% at 1 year, 74.8% at 3 years, 57.2% at 5 years, and 45.8% at 7 years without recurrence in a median period of 1148.5 days The survival was poorer in patients with HCC (47% at 3 years, 9.4% at 5 years, p<0.0001) than without (89.2% at 3 years, 81.9% at 5 years, 67.5% at 7 years), in patients with Child B/C (57.7% at 3 years, 42.1% at 5 years, 28.1% at 7 years, p=0.0016) than with Child A (91.8% at 3 years, 71.5% at 5 years, 62.1% at 7 years), and in patients with GP-R > or = 1.0 (58.9% at 3 years, p=0.0485) than with GP-R<1.0 (76.3% at 3 years, 62% at 5 years, 49.6% at 7 years). Multivariate analysis identified the presence of HCC (hazard ratio, 12.486; 95% CI, 4.08-38.216; p<0.0001), Child B/C (hazard ratio, 3.41; 95% CI, 1.594-7.15; p=0.0051) and GP-R > or = 1.0 (hazard ratio, 2.701; 95% CI, 1.07-6.15; p=0.0221) as independent factors for poor prognosis. CONCLUSION: GP-R > r= 1.0 on Doppler ultrasound before B-RTO may be a predictive indicator for poor prognosis in cirrhotic patients with FV after B-RTO, in addition to the presence of HCC and severe liver damage. PMID- 20004073 TI - An approach to on-line electrospray mass spectrometric detection of polypeptide antibiotics of enramycin for high-speed counter-current chromatographic separation. AB - In the field of pharmaceutical and biomedical analysis of peptides, a rapid on line detection and identification for a methodology have been required for the discovery of new biological active products. In this study, a high-speed counter current chromatography with electrospray mass spectrometry (HSCCC/ESI-MS) was developed for the on-line detection and purification of polypeptide antibiotics of enramycin-A and -B. The analytes were purified on HSCCC model CCC-1000 (multi layer coil planet centrifuge) with a volatile solvent of two-phase system composed of n-butanol/hexane/0.05% aqueous trifluoroacetic acid solution (43/7/50, V/V/V), and detected on an LCMS-2010EV quadrupole mass spectrometer fitted with an ESI source system in positive ionization following scan mode (m/z 100-2000). The HSCCC/ESI-MS peaks indicated that enramycin-A (major m/z 786 [M+3H](3+) and minor m/z 1179 [M+2H](2+)) and enramycin-B (major m/z 791 [M+3H](3+) and minor m/z 1185 [M+2H](2+)) have the peak resolution value of 2.9 from 15mg of loaded enramycin powder. The HSCCC collected amounts of the peak fractions were additionally 4.3mg (enramycin-A), and 5.9mg (enramycin-B), respectively. These purified substances were analyzed by LC/ESI-MS with scan positive mode. Based on the LC/ESI-MS chromatograms and spectra of the fractions, enramycin-A and -B were estimated to be over 95% purity. The overall results indicate that this approach of HSCCC/ESI-MS is a powerful technique for the purification and identification of bioactive peptides. PMID- 20004074 TI - Comparison of ultraviolet detection, evaporative light scattering detection and charged aerosol detection methods for liquid-chromatographic determination of anti-diabetic drugs. AB - Recently, charged aerosol detection (CAD), a new kind of universal detection method, has been widely employed in the HPLC system. In the present study, four kinds of anti-diabetic drug standards, glipizide, gliclazide, glibenclamide and glimepiride were determined by ultraviolet (UV) detection, evaporative light scattering detection (ELSD) and the aforementioned CAD. The results were compared with reference to linearity, accuracy, precision and limit of detection (LOD). All of the experiments were performed on a reverse phase column with water and acetonitrile as the mobile phase. Separations were achieved under the same chromatographic conditions for each detection method. As a result, CAD generated nearly uniform responses compared with UV detection and ELSD. It showed the best accuracy and LOD among 3 detectors and had similar precision with UV detection at higher concentrations while UV detection showed a better precision at lower concentrations than did CAD or ELSD. The LOD of CAD, in fact, can be up to two times higher than that of ELSD. The UV and ELSD linearity was satisfactory at R(2)>0.99, though in the case of CAD, a log-log transformation was needed. The proposed methods were also applied to the real anti-diabetic drugs and diabetes related dietary supplements. PMID- 20004075 TI - Associations between emotional exhaustion, social capital, workload, and latitude in decision-making among professionals working with people with disabilities. AB - BACKGROUND: Many people working in human services in Western countries suffer from burnout, characterized by emotional exhaustion, depersonalization, and decreased personal performance. Prevention of emotional exhaustion (the first phase of burnout) constitutes a great challenge because emotional exhaustion may cause increasing turnover rates in staff and lead to a lesser quality of care. Prevention of emotional exhaustion requires knowledge of its predictors. The aim of this study was to investigate the associations between emotional exhaustion, social capital, workload, and latitude in decision-making among German professionals working in the care of persons with intellectual and physical disabilities. MATERIALS AND METHODS: The study was based on a survey in a sheltered workshop and 5 homes for disabled persons with 175 professionals. Burnout was measured with the German version of the Maslach Burnout Inventory General Survey (MBI-GS). A multivariate logistic regression analysis was computed. RESULTS: Logistic regression identified the following three significant predictors of emotional exhaustion in the sample: workload (OR, 4.192; CI, 2.136 8.227), latitude in decision-making (OR, 0.306; CI, 0.115-0.811), and male gender (OR, 4.123; CI, 1.796-9.462). Nagelkerke's Pseudo-R(2) was 0.344. CONCLUSIONS: The results of this study demonstrate that specific factors in work organization are associated with emotional exhaustion. Taking into account sociodemographic changes and the upcoming challenges for human services professionals, the results underline the importance of considering aspects of organization at the workplace to prevent burnout. Specific circumstances of male employees must be considered. PMID- 20004076 TI - Breast cancer diagnosis in digital mammogram using multiscale curvelet transform. AB - This paper presents an approach for breast cancer diagnosis in digital mammogram using curvelet transform. After decomposing the mammogram images in curvelet basis, a special set of the biggest coefficients is extracted as feature vector. The Euclidean distance is then used to construct a supervised classifier. The experimental results gave a 98.59% classification accuracy rate, which indicate that curvelet transformation is a promising tool for analysis and classification of digital mammograms. PMID- 20004077 TI - Breast cancer survivors and vitamin D: a review. AB - Recent evidence has suggested a role for vitamin D in breast cancer prevention and survival. Studies have reported an inverse relation between vitamin D intake and the risk of breast cancer, improvements in survival after a diagnosis of breast cancer in women with higher levels of vitamin D, and vitamin D insufficiency in up to 75% of women with breast cancer. Preclinical data have indicated that vitamin D affects up to 200 genes that influence cellular proliferation, apoptosis, angiogenesis, terminal differentiation of normal and cancer cells, and macrophage function. Vitamin D receptors have been found in up to 80% of breast cancers, and vitamin D receptor polymorphisms have been associated with differences in survival. Although ongoing studies have investigated a possible link between adequate levels of vitamin D and improved cancer prognosis, breast cancer survivors may derive additional, non-cancer related benefits from adequate vitamin D levels, including improvements in bone mineral density, quality of life, and mood. Maintaining adequate vitamin D stores is recommended for breast cancer survivors throughout their lifetime. PMID- 20004078 TI - Intake of nutritional supplements among people exercising in gyms and influencing factors. AB - OBJECTIVE: To assess supplement intake in people who exercise regularly in gyms in the city of Belo Horizonte, Brazil, and discuss the influencing factors on its ingestion. METHODS: A total of 1102 enrolled subjects who exercised in 50 gyms throughout the city were part of this cross-sectional study. Men and women were recruited and all subjects were at least 18 y old. Participants were asked to complete written questionnaires about their use of supplements. Data were collected over a period of 4 mo. Descriptive statistics and chi-square tests were performed. RESULTS: The intake of nutritional supplements was reported by 36.8% of participants. The highest intake was in men (44.6%). Five products were consumed almost daily: those rich in proteins and amino acids (58%), isotonic drinks (32%), those rich in carbohydrates (23%), natural/phytotherapeutic (20%), and multivitamin/mineral supplements (19%). Most people (55%) reported using nutritional supplements without any specialized professional guidance and based primarily on self-prescription. Individuals younger than 30 y, mainly men (odds ratio 3.28, 95% confidence interval 2.06-5.20, P < 0.01), took supplements rich in proteins (odds ratio 2.73, 95% confidence interval 1.26-5.94, P < 0.05). In contrast, older participants reported taking supplements rich in multivitamin/minerals and natural/phytotherapeutic agents (odds ratio 5.52, 95% confidence interval 2.76-11.06, P < 0.01). CONCLUSION: Our results show that supplement intake in people exercising in gyms is high and is usually self prescribed. We emphasize that the use of dietary supplements must be always done under the supervision of a specialist (physician or nutritionist). PMID- 20004079 TI - Role of enteral nutrition and pharmaconutrients in conditions of splanchnic hypoperfusion. AB - In critically ill patients there is consistent evidence that significant benefits are achieved if nutrients are delivered within the gut compared with the parenteral route. However, in conditions related to gut hypoflux, enteral nutrition may play a double role in counterbalancing the installed low-flow state. On the one hand, enteral-induced postprandial hyperemia may preserve the mucosal barrier and ameliorate immune competence; on the other hand, feeding by the gut may pose a theoretical risk of intestinal ischemia. Despite limited investigation, a strategic temporary minimal enteral nutrition with hypocaloric content has been recommended recently aiming to avoid the overfeeding syndrome and the menace of gut hypoperfusion. Under these conditions, the early luminal delivery of key nutrients such as arginine, glutamine dipeptides, antioxidants, and butyrate are an attractive option for this subset of patients. Arginine may prevent intestinal injury due to hypoperfusion but may harm the gut if ischemia is established. In contrast, glutamine may promote benefits in both conditions. Further investigations by randomized trials in this field are necessary. PMID- 20004080 TI - Body composition changes with aging: the cause or the result of alterations in metabolic rate and macronutrient oxidation? AB - It has been well documented that as individuals age, body composition changes, even in the absence of changes in body weight. Studies have shown that fat mass increases and muscle mass decreases with age. However, it is unclear why such changes occur. Resting metabolic rate (RMR) and substrate oxidation rates have been examined with aging. It has been proposed that reductions in RMR and fat oxidation may lead to changes in body composition. Alternatively, changes in body composition with aging may lead to reductions in RMR. The purpose of this review is to provide an overview of the literature surrounding the impact of aging on RMR and substrate oxidation. Although long-term longitudinal studies are lacking, most cross-sectional studies or short-term longitudinal studies show a reduction in RMR with aging that cannot be explained by changes in body composition including loss in fat-free mass, where the latter includes atrophy or decreases in the mass of high metabolic rate organs. There is indirect evidence suggesting that the metabolic rate of individual organs is lower in older compared with younger individuals. With aging, we conclude that reductions in the mass of individual organs/tissues and in tissue-specific organ metabolic rate contribute to a reduction in RMR that in turn promotes changes in body composition favoring increased fat mass and reduced fat-free mass. PMID- 20004081 TI - Butyrate and propionate induced activated or non-activated neutrophil apoptosis via HDAC inhibitor activity but without activating GPR-41/GPR-43 pathways. AB - OBJECTIVE: Decreased neutrophil apoptosis is implicated in persistent inflammation resulting in systemic inflammatory response syndrome and multiple organ dysfunctions syndromes. Short-chain fatty acids (SCFAs) may be a candidate to control neutrophil apoptosis because SCFAs are normally produced in the gut and related products have been approved for human use. We investigated the effects of SCFAs on apoptosis of activated and non-activated neutrophils and their mechanisms. METHODS: Purified neutrophils obtained from healthy volunteers were preincubated for 1 h with or without the G-protein receptor (GPR) inhibitor pertussis toxin (100 ng/mL) or U-73122 (50 ng/mL), extracellular signal-related protein kinase inhibitor PD98059 (10 microM), mitogen-activated protein kinase (MAPK) p38 inhibitor SB203580 (25 microM), Jun kinase inhibitor-I (2 microM), caspase-3 and -7 inhibitor Z-VAD-FMK (100 microM), caspase-8 inhibitor Z-IETD-FMK (50 microM), or caspase-9 inhibitor Z-LEHD-FMK (50 microM). The cells were then cultured with or without SCFAs or trichostatin A, a typical histone deacetylase inhibitor, in the presence or absence of lipopolysaccharide (1 microg/mL) or tumor necrosis factor-alpha (100 ng/mL). Neutrophil apoptosis was assessed by annexin V staining using flow cytometry. The GPR-41 and -43 and apoptosis-related proteins (bax, mcl-1, a1) mRNA were measured by quantitative real-time polymerase chain reaction and the expression of acetylated histone H3 was determined by western blot. RESULTS: The caspase inhibitors inhibited butyrate- and propionate induced neutrophil apoptosis treated or untreated with lipopolysaccharide or tumor necrosis factor-alpha, whereas GPR and MAPK inhibitors had no effect. The mRNA expressions of GPR-43 and a1 protein were reduced by butyrate and propionate. The expressions of acetylated histone H3 were induced by butyrate and propionate. CONCLUSION: These results suggest that butyrate and propionate increase apoptosis of neutrophils irrespective of their activation state, by factors other than GPRs and MAPKs, and their mechanisms likely relate to their histone deacetylase inhibition activity, which may control a1 mRNA expression. PMID- 20004082 TI - Update on melanocortin interventions for cachexia: progress toward clinical application. AB - Cachexia is a devastating syndrome of body wasting that is associated with multiple common chronic diseases including cancer, chronic kidney disease, and chronic heart failure. These underlying diseases are associated with increased levels of inflammatory cytokines and result in anorexia, increased resting energy expenditure, and loss of fat and lean body mass. Prior experiments have implicated the central melanocortin system in the hypothalamus with the propagation of these symptoms of cachexia. Pharmacologic blockade of this system using melanocortin antagonists causes attenuation of the signs of cachexia in laboratory models. Recent advances in our knowledge of this disease process have involved further elucidation of the pathophysiology of melanocortin activation and demonstration of the efficacy of melanocortin antagonists in new models of cachexia, including cardiac cachexia. In addition, small molecule antagonists of the melanocortin-4 receptor continue to be introduced, including ones with oral bioavailability. These developments generate optimism that melanocortin antagonism will be used to treat humans with disease-associated cachexia. However, to date, human application has remained elusive and it is unclear when we will know whether humans with cachexia would benefit from treatment with these compounds. PMID- 20004084 TI - Influence of natural zeolitic tuff and organozeolites surface charge on sorption of ionizable fumonisin B(1). AB - Natural zeolitic tuff was modified with 2, 5 and 10mmol M(+)/100g of octadecyldimethylbenzyl ammonium (ODMBA) ions and the products were denoted as OZ 2, OZ-5 and OZ-10. The starting material and organozeolites were characterized by determination of the point of zero charge (pH(pzc)) and by thermal analysis. In vitro sorption of fumonisin B(1) (FB(1)) was studied for all sorbents at pH 3, 7 and 9. The pH(pzc) for the zeolitic tuff was 6.8+/-0.1, while the pH(pzc) for OZ 2, OZ-5 and OZ-10 pH(pzc) was 7.0+/-0.1. The curves pH(final)=f(pH(initial)) suggest that the surfaces of all sorbents are positively charged at pH 3 and uncharged at pH 7 and 9. High sorption of FB(1) by the zeolitic tuff in acidic solution suggests electrostatic interactions between the anionic FB(1) and the positively charged surface. At pH 7 and 9, adsorption of FB(1) is prevented because anionic FB(1) cannot be adsorbed at the uncharged surface. From the pH(pzc) for the organozeolites, it is possible that with lower amounts of ODMBA (OZ-2 and OZ-5), at pH 3, beside interactions between head groups of ODMBA and its alkyl chains and anionic FB(1), electrostatic interactions between positive uncovered surface and anionic FB(1) contribute to the sorption, while at pH 7 and 9 there is only the possibility for interactions between FB(1) and ODMBA. When the zeolitic surface was completely covered with ODMBA (OZ-10), FB(1) sorption was independent of the form of FB(1) suggesting only interactions between ODMBA and FB(1). PMID- 20004083 TI - Low omega-6/omega-3 polyunsaturated fatty acid ratios reduce hepatic C-reactive protein expression in apolipoprotein E-null mice. AB - OBJECTIVE: Expression characteristics of C-reactive protein (CRP) for the omega 6/omega-3 polyunsaturated fatty acid (PUFA) ratios have not been evaluated in the well-qualified experimental atherosclerotic mouse model. This work focused on characteristics of CRP expression in the liver of apolipoprotein E-null (apoE(-/ )) mice influenced by omega-6/omega-3 PUFA ratios. METHODS: Varying ratios of omega-6/omega-3 PUFAs (group 1, 1.28; group 2, 5.03; group 3, 9.98; and group 4, 68.26, respectively) on hepatic and aortic CRP expressions were assessed in male apoE(-/-) mice fed a diet containing 5% (w/w) experimental fat for 6 wk. Hepatic peroxisome proliferator-activated receptor-gamma mRNA abundance, hepatic interleukin (IL)-6 protein level, atherosclerotic lesions, and serum cytokines including IL-1beta, IL-6, and tumor necrosis factor-alpha were examined. RESULTS: As the dietary ratio of omega-6/omega-3 fatty acids ascended, so did the expression of hepatic and aortic CRP and hepatic IL-6 protein. However, peroxisome proliferator-activated receptor-gamma mRNA level had a tendency to decrease. Serum IL-1beta, IL-6, and tumor necrosis factor-alpha levels did not show a statistical difference among the mice fed the four ratios of the omega 6/omega-3 PUFA diet. The group 4 mice developed a significant increase in atherosclerotic lesions compared with the other groups. CONCLUSION: The results indicated that low ratios of omega-6/omega-3 PUFAs (1.28-9.98) downregulated the hepatic and aortic CRP expressions and reduced aortic en face lesions in apoE(-/ ) mice compared with the high ratio of the omega-6/omega-3 PUFA diet. PMID- 20004085 TI - Electrophoresis of human brain microvascular endothelial cells with uptake of cationic solid lipid nanoparticles: effect of surfactant composition. AB - This study analyzes the effects of Tween 80 and Span 20 on the electrical interaction between cationic solid lipid nanoparticles (CSLNs) and human brain microvascular endothelial cells (HBMECs). Electrophoretic mobility, zeta potential and fixed charge density of CSLNs and HBMECs with uptake of CSLNs were also investigated. The results revealed that a higher molar ratio of Tween 80 yielded smaller CSLNs, higher charge of CSLNs and larger electrical attraction energy between CSLNs and HBMECs. On the contrary, a decrease in the molar ratio of Tween 80 enhanced the absolute values of electrophoretic mobility and zeta potential of CSLNs-incorporating HBMECs. In addition, a higher concentration of CSLNs in the dispersion medium led to lower charge of CSLNs-incorporating HBMECs. An increased concentration of glutamate from 1 to 2mg/mL enhanced the absolute values of electrophoretic properties of CSLNs-incorporating HBMECs. In the case of increasing concentration of glutamate, pure Tween 80 was stronger in charge enhancement than pure Span 20. PMID- 20004086 TI - Lung lymphatic anatomy and correlates. AB - The pulmonary lymphatics do much more than keep the lung dry. They defend the lungs from airborne particles and microbes and allow a local influx of liquid to clear and clean inflamed or damaged tissue. Lymphatic morphology, especially the three-dimensional structure, is best demonstrated by casting the lymphatics, corroding the tissue, and viewing the casts by scanning electron microscopy. With this technique, different lymphatic forms exist. On the pleural surface prelymphatics are simply tissue planes that connect with lymphatic channels. Reservoir lymphatics are another form of initial lymphatics that have blind ending pouches. They empty into conduit lymphatics. Lymphatics around blood vessels and airways are generally tubular and saccular. In the last decade, lymphatic markers have been discovered that allow the study of lymphatic development in health and disease. Modulators of these pathways could be potential therapeutics for diverse pulmonary problems such as cancer and lung transplantation. The size of the lymphatic system expands manifold in response to an excess fluid load, cancer, or inflammation. Immune cells move through the lymphatics, mature, and become activated there. The lymphatics enable the immune defense system by allowing a sequestered place and close proximity for antigen presentation and lymphocyte maturation. PMID- 20004088 TI - Pulmonary vasodilator testing and use of calcium channel blockers in pulmonary arterial hypertension. AB - Pulmonary arterial hypertension (PAH) encompasses a number of diseases responsible for a specific set of hemodynamic findings during right heart catheterization. During initial workup, pulmonary vasodilator testing is performed. A positive acute pulmonary vasodilator test predicts better survival and response to calcium channel blocker (CCB) therapy. There is lack of consensus on the preferred agent for determining acute pulmonary vasoreactivity. The ACCP guidelines and the 4(th) World Symposium on Pulmonary Hypertension support the use of intravenous epoprostenol or nitric oxide (NO) as the preferred agents for pulmonary vasodilator testing. A decrease in the mean pulmonary artery pressure by at least 10 mmHg to reach an absolute value of 40 mmHg or less without a decrease in cardiac output is currently considered a positive pulmonary vasodilator test. A positive test by the current recommended criteria is observed in about 10-15% of patients with idiopathic PAH. Approximately half of these patients will experience long-term benefits with CCBs. A positive test may select patients with an earlier or less aggressive form of disease, which may carry a better prognosis. A positive vasodilator test is observed very infrequently in patients with pulmonary arterial hypertension other than idiopathic PAH or anorexigen associated PAH. This article reviews the literature regarding pulmonary vasodilator testing and use of CCB therapy in patients with PAH, while identifying the gaps in knowledge concerning this diagnostic procedure. PMID- 20004087 TI - A GMM-IG framework for selecting genes as expression panel biomarkers. AB - OBJECTIVE: The limitation of small sample size of functional genomics experiments has made it necessary to integrate DNA microarray experimental data from different sources. However, experimentation noises and biases of different microarray platforms have made integrated data analysis challenging. In this work, we propose an integrative computational framework to identify candidate biomarker genes from publicly available functional genomics studies. METHODS: We developed a new framework, Gaussian Mixture Modeling-Coupled Information Gain (GMM-IG). In this framework, we first apply a two-component Gaussian mixture model (GMM) to estimate the conditional probability distributions of gene expression data between two different types of samples, for example, normal versus cancer. An expectation-maximization algorithm is then used to estimate the maximum likelihood parameters of a mixture of two Gaussian models in the feature space and determine the underlying expression levels of genes. Gene expression results from different studies are discretized, based on GMM estimations and then unified. Significantly differentially-expressed genes are filtered and assessed with information gain (IG) measures. RESULTS: DNA microarray experimental data for lung cancers from three different prior studies was processed using the new GMM-IG method. Target gene markers from a gene expression panel were selected and compared with several conventional computational biomarker data analysis methods. GMM-IG showed consistently high accuracy for several classification assessments. A high reproducibility of gene selection results was also determined from statistical validations. Our study shows that the GMM-IG framework can overcome poor reliability issues from single-study DNA microarray experiment while maintaining high accuracies by combining true signals from multiple studies. CONCLUSIONS: We present a conceptually simple framework that enables reliable integration of true differential gene expression signals from multiple microarray experiments. This novel computational method has been shown to generate interesting biomarker panels for lung cancer studies. It is promising as a general strategy for future panel biomarker development, especially for applications that requires integrating experimental results generated from different research centers or with different technology platforms. PMID- 20004089 TI - Plasma ammonia response to incremental cycling and walking tests in COPD. AB - OBJECTIVE: It is well documented that plasma ammonia accumulates during exercise under conditions of metabolic stress. Metabolic stress (when skeletal muscle ATP supply fails to meet demand) occurs at low work rates during cycling in patients with COPD, but not been described during walking. Walking is an important activity for many patients with COPD and is commonly prescribed in pragmatic outpatient pulmonary rehabilitation programmes. In this study we explored whether metabolic stress occurs during incremental walking at the low work rates these patients achieve. METHODS: Twenty-nine subjects with stable COPD [mean(SD) age 68(7)years, FEV(1) 50(19)% predicted] performed maximal cardiopulmonary exercise tests on a cycle ergometer and treadmill. Plasma ammonia concentration was measured at rest, 1 and 2min of exercise, peak exercise and 2min recovery. RESULTS: Subjects achieved mean(SD) cycle work rate of 57(20)W with VO(2max) 15.5(4.6)ml/min per kg, and treadmill distance 284(175)m with VO(2peak) 16.8(4.2)ml/min per kg. Plasma ammonia concentration rose significantly (p<0.001) with walking [mean(SEM) change 24.7(3.8)micromol/l] and cycling [mean(SEM) change 35.2(4.3)micromol/l], but peak exercise ammonia was lower in walking (p<0.01). In a subgroup of subjects (n=7) plasma ammonia did not rise during either cycling or walking despite similar lactate rise and peak exercise indices. CONCLUSION: Our data indicate that failure of muscle ATP re-synthesis to meet demand and development of metabolic stress can occur during walking in COPD patients at the low work rates these patients achieve. This may therefore be a factor contributing to exercise limitation independent of ventilatory limitation. PMID- 20004090 TI - Emitted dose and lung deposition of inhaled terbutaline from Turbuhaler at different conditions. AB - Turbuhaler has a very high resistance hence patient inhalation flow when using it would be low. The total emitted dose (TED) of 500microg terbutaline sulphate from a Bricanyl Turbuhaler was determined using a range of inhalation flows (10-60L min(-1)) with inhalation volume of 2 and 4L using a DPI sampling apparatus after one and two inhalations. The relative lung and systemic bioavailability of terbutaline from Bricanyl Turbuhaler when used by healthy subjects and COPD patients were determined after one and two inhalations at slow and fast inhalation flows using a novel urinary terbutaline pharmacokinetic method. The TED resulted from the one and two inhalations increased significantly (p<0.05) with the increase of the inhalation flow at both 2 and 4L inhalation volumes. The relative lung and systemic bioavailability after one inhalation at fast inhalation flow were significantly higher (p<0.01) than at slow inhalation flow in both healthy subjects and patients. Also the healthy subjects results were significantly higher (p<0.05) than the COPD patients after one inhalation. However after two inhalations there was no significant difference between slow and fast inhalation flow or healthy subjects and COPD patients. Hence it is essential to inhale twice and as deep and hard as possible from each dose of Turbuhaler for patients with low inspiratory flow and limited inhalation volume as they may not receive much benefit from one inhalation. PMID- 20004091 TI - A portable microfluidic flow cytometer based on simultaneous detection of impedance and fluorescence. AB - A portable microfluidic flow cytometer with dual detection ability of impedance and fluorescence was developed for cell analysis and particle-based assays. In the proposed system, fluorescence from microparticles and cells is measured through excitation by a light emitting diode (LED) and detection by a solid stated photomultiplier (SSPM). Simultaneous impedometric detection provides information on the existence and size of microparticles and cells through polyelectrolyte gel electrodes (PGEs) operated by custom designed circuits for signal detection, amplification, and conversion. Fluorescence and impedance signals were sampled at 1 kHz with 12 bit resolution. The resulting microfluidic cytometer is 15x10x10 cm(3) in width, depth, and height, with a weight of about 800 g. Such a miniaturized and battery powered system yielded a portable microfluidic cytometer with high performance. Various microbeads and human embryonic kidney 293 (HEK-293) cells were employed to evaluate the system. Impedance and fluorescence signals from each bead or cell made classification of micro particles or cells easy and fast. PMID- 20004092 TI - Ethanol production from high dry matter corncob using fed-batch simultaneous saccharification and fermentation after combined pretreatment. AB - To obtain high concentration of ethanol from cellulose, corncob was pretreated with acid and alkali to remove non-cellulose components, and then subjected to simultaneous saccharification and fermentation (SSF). An ethanol concentration as high as 69.2 g/L was achieved with 19% dry matter (DM) using batch SSF, resulting in an 81.2% overall ethanol yield. A fed-batch process using a high solid concentration was also investigated. Fresh substrate was pretreated with dilute sulfuric acid-sodium hydroxide, and then added at different amounts during the first 24 h, to yield a final dry matter content of 25% (w/v). SSF conditions with cellulose loading of 22.8 FPU/g glucan, dry yeast (Saccharomyces cerevisiae) loading of 5 g/L and substrate supplementation every 4h yielded the highest ethanol concentration of 84.7 g/L after 96 h. This corresponded to a 79% overall ethanol yield. PMID- 20004093 TI - Biodegradation of the analgesic naproxen by Trametes versicolor and identification of intermediates using HPLC-DAD-MS and NMR. AB - The white-rot fungus Trametes vesicolor degraded naproxen (10 mg L(-1)) in a liquid medium to non-detectable levels after 6h. When naproxen was added in the range of concentrations typically found in the environment (55 microg L(-1)), it was almost completely degraded (95%) after 5h. In vitro degradation experiments with purified laccase and purified laccase plus mediator 1-hydroxybenzotriazol showed slight and almost complete naproxen degradation, respectively. A noticeable inhibition on naproxen degradation was also observed when the cytochrome P450 inhibitor 1-aminobenzotriazole was added to the fungal cultures. These data suggest that both enzymatic systems could play a role in naproxen degradation. 2-(6-hydroxynaphthalen-2-yl)propanoic acid and 1-(6 methoxynaphthalen-2-yl)ethanone were structurally elucidated by HPLC-DAD-MS and NMR as degradation intermediates of naproxen. After 6h of incubation, both parent compound and intermediates disappeared from the medium. The non-toxicity of the treated medium was confirmed by Microtox test. PMID- 20004094 TI - Two-step liquid hot water pretreatment of Eucalyptus grandis to enhance sugar recovery and enzymatic digestibility of cellulose. AB - A two-step liquid hot water pretreatment (TSLHW) was developed with the objective of achieving complete saccharification of both hemicellulose and cellulose of Eucalyptus grandis, thereby avoiding the problems associated with the use of strong acid catalysts. The first step of the pretreatment was studied in the temperature range 180-200 degrees C, and the highest yield of total xylose achieved was 86.4% after 20 min at 180 degrees C. The second-step of the pretreatment was studied in the temperature range 180-240 degrees C and for lengths of time of 0-60 min. The conversion rate of glucan was more sensitive to temperature than time. The optimum reaction conditions for the second step of the pretreatment with minimal degradation of sugars were 200 degrees C for 20 min. the total sugar recovery from E. grandis with the optimized pretreatment and 72 h enzymatic digestion, reached 96.63%, which is superior to the recovery from a single-step pretreatment with hot water or dilute acid. PMID- 20004095 TI - Wood plastic composites from agro-waste materials: Analysis of mechanical properties. AB - This article presents the application of agro-waste materials (i.e., corn stalk, reed stalk, and oilseed stalk) in order to evaluate and compare their suitability as reinforcement for thermoplastics as an alternative to wood fibers. The effects of fiber loading and CaCO(3) content on the mechanical properties were also studied. Overall trend shows that with addition of agro-waste materials, tensile and flexural properties of the composites are significantly enhanced. Oilseed fibers showed superior mechanical properties due to their high aspect ratio and chemical characteristics. The order of increment in the mechanical properties of the composites is oilseed stalk >corn stalk>reed stalk at all fiber loadings. The tensile and flexural properties of the composite significantly decreased with increasing CaCO(3) content, due to the reduction of interface bond between the fiber and matrix. It can be concluded from this study that the used agro-waste materials are attractive reinforcements from the standpoint of their mechanical properties. PMID- 20004096 TI - Identification of a new class of small molecule C5a receptor antagonists. AB - C5a is a terminal product of the complement cascade that activates and attracts inflammatory cells including granulocytes, mast cells and macrophages via a specific GPCR, the C5a receptor (C5aR). Inhibition of C5a/C5aR interaction has been shown to be efficacious in several animal models of autoimmune diseases, including RA, SLE and asthma. This account reports the discovery of a new class of C5aR antagonists through high-throughput screening. The lead compounds in this series are selective and block C5a binding, C5a-promoted calcium flux in human neutrophils with nanomolar potency. PMID- 20004097 TI - Synthesis and biological activity of a potent and orally bioavailable SCD inhibitor (MF-438). AB - A series of stearoyl-CoA desaturase 1 (SCD1) inhibitors were developed. Investigations of enzyme potency and metabolism led to the identification of the thiadiazole-pyridazine derivative MF-438 as a potent SCD1 inhibitor. MF-438 exhibits good pharmacokinetics and metabolic stability, thereby serving as a valuable tool for further understanding the role of SCD inhibition in biological and pharmacological models of diseases related to metabolic disorders. PMID- 20004098 TI - Synthesis, spectral analysis and in vitro microbiological evaluation of 3-(3 alkyl-2,6-diarylpiperin-4-ylidene)-2-thioxoimidazolidin-4-ones as a new class of antibacterial and antifungal agents. AB - In the present work, a new series of bis hybrid heterocycle comprising both piperidine and thiohydantoin nuclei together namely 3-(3-alkyl-2,6-diarylpiperin 4-ylidene)-2-thioxoimidazolidin-4-ones 46-60 was synthesized by the treatment of the respective thiosemicarbazones 31-45 with chloroethyl acetate and anhydrous sodium acetate in refluxing ethanol for 4h and were characterized by melting point, elemental analysis, MS, FT-IR, one-dimensional NMR ((1)H, D(2)O exchanged (1)H and (13)C), two dimensional HOMOCOSY and NOESY spectroscopic data. In addition, the title compounds were screened for their antimicrobial activities against a spectrum of clinically isolated microbial organisms. Compounds 47-50, 52-55 and 57-60 with fluoro, chloro, methoxy or methyl functions at the para position of phenyl rings attached to C-2 and C-6 carbons of piperidine moiety along with and without methyl substituent at position C-3 of the piperidine ring exerted potent biological activities against Staphylococcus aureus, beta Hemolytic streptococcus, Vibrio cholerae, Escherichia coli, Pseudomonas aeruginosa, Aspergillus flavus, Candida albicans, Candida 6 and Candida 51 at a minimum inhibitory concentration. PMID- 20004099 TI - Evaluation of secondary amide replacements in a series of CCR5 antagonists as a means to increase intrinsic membrane permeability. Part 1: Optimization of gem disubstituted azacycles. AB - Replacement of a secondary amide with an N-acyl or N-sulfonyl gem-disubstituted azacyle in a series of CCR5 antagonists led to the identification of compounds with excellent in vitro HIV antiviral activity and increased intrinsic membrane permeability. PMID- 20004100 TI - Hemorrhagic encephalitis associated with Epstein-Barr virus infection. AB - Epstein-Barr virus (EBV) encephalitis is a rare neurological complication, usually only reported in pediatric patients. We present a 20-year-old, previously healthy male who developed hemorrhagic encephalitis caused by EBV. He was admitted to our hospital with a 1-week history of fever, diarrhea, headache, and confusion. Brain T2-weighted MRI showed a focal area of increased signal in the right temporal lobe. Brain MRI and CT scans on day 2 revealed progression of the lesion, with partial hemorrhagic change, acute brain swelling, and severe midline shift. The patient underwent external decompression and external ventricular drainage. EBV DNA was identified in brain biopsy specimens by polymerase chain reaction. The postoperative course was uneventful. To our knowledge, this is the second report of hemorrhagic EBV encephalitis in an adult. PMID- 20004101 TI - Risk of growth in unruptured intracranial aneurysms: a retrospective analysis. AB - This study sought to define the growth of unruptured intracranial aneurysms, in particular the frequency of growth and the characteristic factors predictive of growth. Two hundred and eight patients with 285 unruptured aneurysms were followed. Electronic records and angiographic films were obtained for measurements of aneurysm size. The mean follow-up duration was 21.8 months (range 1.1-137.3 months). Growth was identified in 95 of the 285 aneurysms (33.3%). The cumulative incidence of growth predicted using the Kaplan-Meier method was 22.7% at 1 year, 35.2% at 2 years, and 47.7% at 3 years. Aneurysm growth was significantly associated with a patient history of excessive alcohol consumption (p=0.04). A high incidence of growth can be seen in conservatively managed aneurysms with time. Consequently, continual follow-up is recommended to monitor for aneurysmal growth. PMID- 20004102 TI - Cerebral inflammatory response and predictors of admission clinical grade after aneurysmal subarachnoid hemorrhage. AB - Poor admission clinical grade is the most important determinant of outcome after aneurysmal subarachnoid hemorrhage (aSAH); however, little attention has been focused on independent predictors of poor admission clinical grade. We hypothesized that the cerebral inflammatory response initiated at the time of aneurysm rupture contributes to ultra-early brain injury and poor admission clinical grade. We sought to identify factors known to contribute to cerebral inflammation as well as markers of cerebral dysfunction that were associated with poor admission clinical grade. Between 1997 and 2008, 850 consecutive SAH patients were enrolled in our prospective database. Demographic data, physiological parameters, and location and volume of blood were recorded. After univariate analysis, significant variables were entered into a logistic regression model to identify significant associations with poor admission clinical grade (Hunt-Hess grade 4-5). Independent predictors of poor admission grade included a SAH sum score >15/30 (odds ratio [OR] 2.3, 95% confidence interval [CI] 1.5-3.6), an intraventricular hemorrhage sum score >1/12 (OR 3.1, 95% CI 2.1-4.8), aneurysm size >10mm (OR 1.7, 95% CI 1.1-2.6), body temperature 38.3 degrees C (OR 2.5, 95% CI 1.1-5.4), and hyperglycemia >200mg/dL (OR 2.7, 95% CI 1.6-4.5). In a large, consecutive series of prospectively enrolled patients with SAH, the inflammatory response at the time of aneurysm rupture, as reflected by the volume and location of the hemoglobin burden, hyperthermia, and perturbed glucose metabolism, independently predicts poor admission Hunt-Hess grade. Strategies for mitigating the inflammatory response to aneurysmal rupture in the hyper-acute setting may improve the admission clinical grade, which may in turn improve outcomes. PMID- 20004103 TI - The influence of race on outcome following subarachnoid hemorrhage. AB - The goal of this study was to examine the relationship between race and outcome following subarachnoid hemorrhage (SAH). We identified all SAH discharges in New York City during 2003. An adverse outcome was defined as in-hospital death or discharge other than to home. While correcting for age and gender, we examined the effect of race and payor status on outcome following SAH. Forty-four percent of patients with SAH were white. Being white had a significant relationship with outcome when controlled for payor status (odds ratio 0.56). Among self pay/Medicaid patients, fewer white (52%) individuals suffered poor outcomes than non-white (66%, p=0.03). Our results establish that white patients in New York City with SAH have better outcomes than non-whites. While it is unclear whether this discrepancy is secondary to pathophysiological differences or unidentified social factors, our findings demonstrate that this effect is independent of insurance status, and emphasize the need for further investigation into racial disparities in outcome following SAH. PMID- 20004104 TI - Custom-made hinged spacers in revision knee surgery for patients with infection, bone loss and instability. AB - Polymethyl methacrylate spacers are commonly used during staged revision knee arthroplasty for infection. In cases with extensive bone loss and ligament instability, such spacers may not preserve limb length, joint stability and motion. We report a retrospective case series of 19 consecutive patients using a custom-made cobalt chrome hinged spacer with antibiotic-loaded cement. The "SMILES spacer" was used at first-stage revision knee arthroplasty for chronic infection associated with a significant bone loss due to failed revision total knee replacement in 11 patients (58%), tumour endoprosthesis in four patients (21%), primary knee replacement in two patients (11%) and infected metalwork following fracture or osteotomy in a further two patients (11%). Mean follow-up was 38 months (range 24-70). In 12 (63%) patients, infection was eradicated, three patients (16%) had persistent infection and four (21%) developed further infection after initially successful second-stage surgery. Above knee amputation for persistent infection was performed in two patients. In this particularly difficult to treat population, the SMILES spacer two-stage technique has demonstrated encouraging results and presents an attractive alternative to arthrodesis or amputation. PMID- 20004105 TI - Improving the trade-off between simulation time and accuracy in efficiency calibrations with the code DETEFF. AB - Quick and relatively simple procedures were incorporated into the Monte Carlo code DETEFF in order to consider the escape of Bremsstrahlung radiation and secondary electrons. The relative bias in efficiency values was thus reduced for photon energies between 1500 and 2000 keV, without any noticeable increment of the simulation time. A relatively simple method was also included to consider the rounding of detector edges. The validation studies showed relative deviations of about 1% in the energy range 10-2000 keV. PMID- 20004106 TI - New coupling strategy for radionuclide labeling of synthetic polymers. AB - We have developed a radiolabeling strategy for synthetic polymers based on the formation of azo dye usable for both covalent and chelating labeling modalities under mild conditions. Poly[N-(2-hydroxypropyl)methacrylamide] and poly(N isopropyl acrylamide) were used as model polymers. N-methacryloyl tyrosinamide was introduced into the polymers and the phenolic moiety was then reacted with diazotized chelator precursors. The conjugates were radiolabeled with both the covalently bound (iodine-125) and chelated (indium-111) radionuclides in high yields and sufficient in vitro stability of the labels was proven. PMID- 20004107 TI - Proteolytic activity and cysteine protease expression in wheat leaves under severe soil drought and recovery. AB - The involvement of acidic proteases in soil drought response of winter wheat (Triticum aestivum L.) at seedling stage in three cultivars differing in water stress tolerance was studied. Withholding irrigation for seven days resulted in severe drought stress corresponding to 60% leaf water deficit. Stressed plants were recovered by providing optimal water supply for 3 days. Reversible changes in leaf pigment and protein content were registered, being least expressed in the drought-resistant cultivar Katya. Protein loss was inversely related to the increase in total proteolytic activity at pH 5 and in aminopeptidase activity at pH 7. Quantitative differences among the cultivars were established only for azocaseinolytic activity (pH 5). The drought-resistant cultivar (Katya) showed relatively little increase in acid protease activity whereas the highest values of this activity were detected in cultivar Pobeda. In-gel staining for cysteine activated proteases revealed four to five separate activity bands. The upper band, specifically inhibited by E-64, was raised at severe drought. Transcript abundance of two wheat cysteine proteases -Ta.61026 putative thiol protease, and WCP2 peptidase of papain type was analyzed by RT-PCR. Gene expression of the cysteine proteases under study was suppressed in the drought-tolerant cultivar, while in the less resistant ones it remained unchanged or augmented. The results suggest that lower proteolytic activity and decreased expression of certain cysteine protease genes under water deficit during early developmental stage could be regarded as an indicator for drought resistance of winter wheat cultivars. PMID- 20004108 TI - Differential survival of Escherichia coli uvrA, uvrB, and uvrC mutants to psoralen plus UV-A (PUVA): Evidence for uncoupled action of nucleotide excision repair to process DNA adducts. AB - The nucleotide excision repair mechanism (NER) of Escherichia coli is responsible for the recognition and elimination of more than twenty different DNA lesions. Herein, we evaluated the in vivo role of NER in the repair of DNA adducts generated by psoralens (mono- or bi-functional) and UV-A light (PUVA) in E. coli. Cultures of wild-type E. coli K12 and mutants for uvrA, uvrB, uvrC or uvrAC genes were treated with PUVA and cell survival was determined. In parallel, kinetics of DNA repair was also evaluated by the comparison of DNA sedimentation profiles in all the strains after PUVA treatment. The uvrB mutant was more sensitive to PUVA treatment than all the other uvr mutant strains. Wild-type strain, and uvrA and uvrC mutants were able to repair PUVA-induced lesions, as seen by DNA sedimentation profiles, while the uvrB mutant was unable to repair the lesions. In addition, a quadruple fpg nth xth nfo mutant was unable to nick PUVA-treated DNA when the crude cell-free extract was used to perform plasmid nicking. These data suggest that DNA repair of PUVA-induced lesions may require base excision repair functions, despite proficient UvrABC activity. These results point to a specific role for UvrB protein in the repair of psoralen adducts, which appear to be independent of UvrA or UvrC proteins, as described for the classical UvrABC endonuclease mechanism. PMID- 20004109 TI - Brain metastases in HER2-positive breast cancer: the evolving role of lapatinib. AB - Due to improvements in diagnosis and systemic therapy, brain metastases are an increasingly common cause of morbidity and mortality for patients with advanced breast cancer. The incidence of symptomatic brain metastases among women with metastatic breast cancer ranges from 10% to 16%. The HER2 receptor, which is overexpressed in approximately 25% of all breast cancers, is an important risk factor for the development of central nervous system metastases. Surgery and radiation therapy are the primary approaches to the treatment of brain metastases but new chemotherapy and biological agents promise to play an important role in the future management of central nervous system disease. This article reviews the epidemiology, current treatment options and recent advances in the field, with a focus on HER2-positive disease and the emerging role of lapatinib for the treatment and prevention of brain metastases. PMID- 20004110 TI - Metabolic reprogramming, caloric restriction and aging. AB - Caloric restriction (CR) without malnutrition slows the aging process and extends lifespan in diverse species by unknown mechanisms. The inverse linear relationship between calorie intake and lifespan suggests that regulators of energy metabolism are important in the actions of CR. Studies in several species reveal tissue-specific changes in energy metabolism with CR and suggest that metabolic reprogramming plays a critical role in its mechanism of aging retardation. We herein describe common signatures of CR and suggest how they can slow aging. We discuss recent advances in understanding the function of key metabolic regulators that probably coordinate the response to altered nutrient availability with CR and how the pathways they regulate can retard the aging process. PMID- 20004111 TI - Low-dose erythropoietin aggravates endotoxin-induced organ damage in conscious rats. AB - Endotoxin shock can induce the production of several inflammatory mediators such as TNF-alpha, IL-6, and IL-1beta, leading to multiple organ dysfunction and death. Erythropoietin (EPO) has been found to interact with its receptor (EPO-R), expressed in a wide variety of non-hematopoietic tissues, to induce a range of pleiotropic cytoprotective actions. We investigated the effects of low doses of EPO (300U/kg, intravenous administration) on the physiopathology and cytokine levels in endotoxin shock in conscious rats. Endotoxin shock was induced by intravenous injection of Escherichia coli lipopolysaccharide (20mg/kg) in conscious rats. Mean arterial pressure (MAP) and heart rate (HR) were continuously monitored for 48h after LPS administration. Levels of biochemical and cytokine parameters, including glutamic oxaloacetic transaminase (GOT), glutamic pyruvic transaminase (GPT), blood urea nitrogen (BUN), creatinine (Cre), lactic dehydrogenase (LDH), and creatine phosphokinase (CPK) were measured at 0, 1, 3, 6, 9, 12, 18, 24, and 48h after sepsis. Serum TNF-alpha, IL-6, and IL-1beta level was measured at 1h after sepsis. Endotoxin shock significantly increased blood GOT, GPT, BUN, Cre, LDH, CPK, TNF-alpha, IL-6, IL-1beta levels, and HR, while it decreased MAP. EPO further increased the markers of organ injury (GOT, GPT, BUN, Cre, LDH, and CPK), inflammatory biomarkers (TNF-alpha, IL-6, and IL 1beta) and did not affect MAP and HR after LPS. EPO disserved endotoxin shock induced liver, kidney, lung, and small intestine damage in conscious rats. In conclusion, pre-treatment with low doses of EPO increased the release of TNF alpha, IL-6, and IL-1beta, along with aggravating endotoxin shock-induced markers of organ injury in conscious rats. PMID- 20004112 TI - Interferon gamma gene +874A/T polymorphism and intracellular interferon gamma expression in pulmonary tuberculosis. AB - We investigated whether IFN-gamma gene +874(A/T) polymorphism influences intracellular interferon gamma expression in T-cell subsets of normal healthy subjects (NHS) and pulmonary tuberculosis patients (PTB). Peripheral blood mononuclear cells were stimulated with live Mycobacterium tuberculosis (MTB) and the intracellular IFN-gamma expression was studied using flow cytometry. Genotyping of IFN-gamma gene +874(A/T) was done using allele specific polymerase chain reaction. Significantly increased IFN-gamma expressing CD3+CD4+ and CD3+CD8+ T cells were observed in NHS with AA genotype compared to TT genotype in unstimulated (p=0.0308 and p=0.0157) and MTB stimulated (p=0.0494 and p=0.0287) cultures and this difference was not observed in PTB patients. The present study suggests that the variant genotypes of IFN-gamma (+874) may be associated with altered expression of IFN-gamma at the intracellular level and play an immunoregulatory role at the site of M. tuberculosis infection. PMID- 20004113 TI - The change of cytokines in tear and blood after different pterygium operation. AB - Pterygium is an invasion of altered ocular tissue into the cornea. Bone marrow derived stem cells have been reported to be involved in wound healing under chemotactic factors after pterygium removal and pain may act as a trigger signal. We evaluated the change of systemic and local chemotactic factors that could affect the mobilization and migration of BMSCs to the wound bed after conventional bare sclera pterygium excision. We also applied temporary amniotic membrane patch after pterygium removal, and compared the changes of cytokines with those of conventional bare sclera excision group. Substance-P (SP), vascular endothelial growth factor (VEGF), and stem cell factor (SCF) were measured in plasma and tear using ELISA and migrating CD34(+) cells by flow cytometry. The results showed that post-operative pain was much reduced (p<0.05), and SP, VEGF and SCF kept consistently lower levels in plasma after temporary amniotic membrane application. Circulating CD34(+) cells increased slightly in the temporary amniotic membrane patch group compared with marked increase in the bare sclera group. Thus, the application of a temporary amniotic membrane after pterygium removal might be an effective therapeutic means by controlling pain and excessive infiltration of bone marrow-derived stem cells. PMID- 20004114 TI - Electron capture dissociation mass spectrometry of metallo-supramolecular complexes. AB - The electron capture dissociation (ECD) of metallo-supramolecular dinuclear triple-stranded helicate Fe(2)L(3)(4+) ions was determined by Fourier transform ion cyclotron resonance mass spectrometry. Initial electron capture by the di iron(II) triple helicate ions produces dinuclear double-stranded complexes analogous to those seen in solution with the monocationic metal centers Cu(I) or Ag(I). The gas-phase fragmentation behavior [ECD, collision-induced dissociation (CID), and infrared multiphoton dissociation (IRMPD)] of the di-iron double stranded complexes, (i.e., MS(3) of the ECD product) was compared with the ECD, CID, and IRMPD of the Cu(I) and Ag(I) complexes generated from solution. The results suggest that iron-bound dimers may be of the form Fe(I)(2)L(2)(2+) and that ECD by metallo-complexes allows access, in the gas phase, to oxidation states and coordination chemistry that cannot be accessed in solution. PMID- 20004115 TI - Recognition of additional roles for immunoglobulin domains in immune function. AB - Characterization of immune receptors found in phylogenetically disparate species at the genetic, structural and functional levels has provided unique insight into the evolutionary acquisition of immune function. The roles of variable- and intermediate-type immunoglobulin (Ig) domains in direct recognition of ligands and other functions are far wider than previously anticipated. Common mechanisms of multigene family diversification and expansion as well as unique adaptations that relate to function continue to provide unique insight into the numerous patterns, processes and complex interactions that regulate the host response to infectious challenge. PMID- 20004116 TI - Those other mammals: the immunoglobulins and T cell receptors of marsupials and monotremes. AB - This review summarizes analyses of marsupial and monotreme immunoglobulin and T cell receptor genetics and expression published over the past decade. Analyses of recently completed whole genome sequences from the opossum and the platypus have yielded insight into the evolution of the common antigen receptor systems, as well as discovery of novel receptors that appear to have been lost in eutherian mammals. These species are also useful for investigation of the development of the immune system in organisms notable for giving birth to highly altricial young, as well as the evolution of maternal immunity through comparison of oviparous and viviparous mammals. PMID- 20004117 TI - Measuring "intuition" in the SRT generation task. AB - We address some concerns related to the use of post-trial attribution judgments, originally developed for artificial grammar learning (AGL), during the version of the serial reaction time (SRT) task used by Fu, Dienes, and Fu (2010). In particular, intuition attributions, which are central to Fu et al.'s arguments, seem problematic: This attribution is likely to be made when stimuli contain several competing sources of information to which subjective feelings could be attributed. The interpretation of intuition attributions in Fu et al.'s SRT generation task is problematic because the procedure involved a 2-element sequence where items varied only in position. In our view, responses categorised as intuitions might have been a variety of guess response where neither judgement knowledge nor structural knowledge were conscious. The results would then be compatible with previous findings showing that people can control the use of unconscious structural knowledge even when judgement knowledge is unconscious. PMID- 20004118 TI - The imbalance of oculomotor capture in unilateral visual neglect. AB - Visual neglect has been associated with an imbalance in the level of activity in the saccadic system: activity in the contralesional field is suppressed, which makes target selection unlikely. We recorded eye movements of a patient with hemispatial neglect and a group of healthy participants during an oculomotor distractor paradigm. Results showed that the interfering effects of a distractor were very strong when presented in her ipsilesional visual field. However, when the distractor was presented in her contralesional field, there were no interfering effects when the target was presented in her ipsilesional field. These findings could not be explained by the presence of a visual field defect as revealed by the results of two hemianopic patients. Our results are in line with an imbalance in the level of activity in the saccadic system in visual neglect because visual elements presented in the contralesional field did not compete for saccadic selection. PMID- 20004119 TI - Reproducibility of lung and lobar volume measurements using computed tomography. AB - RATIONALE AND OBJECTIVES: Lung and lobar volume measurements from computed tomographic (CT) imaging are being used in clinical trials to assess new minimally invasive emphysema treatments aiming to reduce lung volumes. Establishing the reproducibility of lung volume measurements is important if they are to be accepted as treatment planning and outcome variables. The aims of this study were to (1) investigate the correlation between lung volumes assessed on CT imaging and on pulmonary function testing (PFT), (2) compare the two methods' reproducibility, and (3) assess the reproducibility of CT lobar volumes. MATERIALS AND METHODS: CT imaging and body plethysmography were performed at baseline and after a 9-month interval in multicenter emphysema treatment trials. Lung volumes were measured at total lung capacity (TLC) and at residual volume (RV). Lobar volumes were measured on CT imaging using a semiautomated technique. The correlations between CT and PFT volumes were computed for 486 subjects at baseline. Reproducibility was assessed in terms of the intraclass correlation coefficient (ICC) for 126 subjects from the control group at TLC and 120 subjects at RV. RESULTS: Correlations between CT and PFT lung volumes were 0.86 at TLC and 0.67 at RV. At TLC, the ICCs were 0.943 for CT imaging and 0.814 for PFT. At RV, the ICCs were 0.886 for CT imaging and 0.683 for PFT. CT lobar volumes showed good reproducibility (all P values < .05). CONCLUSION: CT lung and lobar volume measurements could be captured in a multicenter trial setting with high reproducibility and were highly correlated with those obtained on PFT. CT imaging showed significantly better reproducibility than PFT between interval lung volume measurements, offering the potential for designing emphysema treatment trials involving fewer subjects. PMID- 20004120 TI - Beneficial changes of serum calcification markers and contralateral carotid plaques echogenicity after combined carotid artery stenting plus intensive lipid lowering therapy in patients with bilateral carotid stenosis. AB - OBJECTIVES/DESIGN: In symptomatic patients treated with ipsilateral carotid artery stenting (CAS) plus intensive lipid lowering, we assessed the changes of osteopontin (OPN), osteoprotegerin (OPG) and the Gray-Scale Median (GSM) score contralateral to symptomatic carotid stenosis. MATERIALS/METHODS: Forty-six symptomatic patients (group A) with significant carotid stenosis (North American Symptomatic Carotid Endarterectomy Trial (NASCET): >70%) underwent ipsilateral CAS. Those patients had simultaneously contralateral low-grade carotid stenosis (NASCET: 30-69%). Group B included 67 symptomatic patients with low-grade bilateral carotid stenosis (NASCET: 30-69%), but without indications for revascularisation. All patients were treated with atorvastatin (10-80mg) to target low-density lipoprotein (LDL)<100mgdl(-1). Blood samples and plaques' GSM score contralateral to brain infarct were assayed at baseline and after 6 months. RESULTS: At baseline, there were no significant differences between groups (p>0.05). Six-month atorvastatin treatment equivalently improved lipid profile in both groups (p<0.05). The parameters hsCRP, OPN and OPG were significantly down regulated within both groups, but to a greater extent in group A (p<0.05). Besides this, contralateral GSM score was significantly improved from baseline in both groups (p<0.01), but that increment was more pronounced in group A (vs. group B; p=0.041). These changes were inversely correlated with changes in OPN (p=0.014), OPG (p=0.011) and LDL (p=0.041). CONCLUSION: Ipsilateral CAS plus intensive lipid-lowering therapy was associated with enhanced contralateral carotid plaque stability and attenuated inflammatory burden and calcification inhibitors to a greater extent than atorvastatin therapy alone in patients with bilateral carotid stenosis. PMID- 20004121 TI - The association between acquired urinary sphincter mechanism incompetence in bitches and early spaying: a case-control study. AB - A case-control study was conducted between December 2005 and August 2006 in London (1) to estimate the strength of association between early ovariohysterectomy (spaying) and urinary incontinence (sphincter mechanism incompetence), (2) to identify other risk factors for incontinence, and (3) to assess any implications of incontinence on the owner-pet relationship. Cases were defined as bitches that developed incontinence after spaying and were treated, and the controls comprised continent spayed bitches. Questionnaires from 202 cases were compared to 168 controls, and analysed using multivariable logistic regression. No significant association between early spaying and incontinence was detected although there was a tendency that early spayed bitches were less likely to be incontinent. Docked bitches were 3.8 times more likely to be incontinent than undocked bitches; bitches weighing over 10 kg were 3.7 times more likely to be incontinent than smaller dogs; and older bitches were more likely to be incontinent (OR=3.1-23.8) than younger animals. Some owners were found to have a negative attitude towards incontinence. PMID- 20004122 TI - Reduction of B(0) inhomogeneity effects in triple-quantum-filtered sodium imaging. AB - Triple-quantum (TQ) filtered sodium MR imaging has been proposed for separation of sodium signal arising from different physiological compartments. In a three pulse sequence without refocussing pulse, the TQ signal is strongly sensitive to inhomogeneities of the B(0) field. We examine the dependence of the TQ signal intensity on the sequence parameters and propose a modified phase-cycling scheme to improve image quality. A new method for correction of B(0) inhomogeneity artefacts in TQ filtered sodium imaging is presented which requires only two acquisitions to obtain a correction as far as the B(0) inhomogeneity and the pulse widths are not too large. The method was verified in phantom experiments. PMID- 20004123 TI - IFI = invasive fungal infections. What is that? A misnomer, because a non invasive fungal infection does not exist! AB - In principle, one has to differentiate between a mere colonization and an invasive fungal infection (IFI) with quite different clinical manifestations and consequences. If the term invasive were used in its proper sense (expressis verbis), all fungal infections could be called invasive, even a fungal keratitis or an infection of the hairs by dermatophytes. In general, however, the term IFI is used only to characterize systemic, generalized, deep-seated, visceral and severe, life-threatening fungal infections, in contrast to superficial, local, benign, self-limiting fungal diseases. The term IFI as used generally is thus misleading, confusing, and not at all helpful to correctly differentiate mild, moderate, and severe fungal infections. PMID- 20004124 TI - Human bacterial arthritis caused by Streptococcus zooepidemicus: report of a case. AB - Septic arthritis caused by Streptococcus zooepidemicus is a rare event in humans. Of the four cases reported in the literature, only two patients had direct animal contact, and the portal of entry remained unclear in all cases. We report herein the case of a patient who suffered a purulent arthritis of the left shoulder caused by S. zooepidemicus, successfully treated in our department. A diagnostic FDG-PET-CT scan ruled out other foci of infection, but detected a hyperkeratotic plantar chronic soft tissue lesion of the left foot, acquired in a paragliding accident 10 years earlier. The fact that the patient habitually took care of his horses barefoot in boots, identifies the cutaneous portal of entry as most likely. To our knowledge this is the first report of a septic arthritis caused by S. zooepidemicus where a cutaneous entry route is described. PMID- 20004125 TI - [Hypofractionated stereotactic radiotherapy for brain metastases]. AB - PURPOSE: A survey of the literature has been performed to find arguments in order to help the choice between radiosurgery and hypofractionated stereotactic radiotherapy in the treatment of brain metastases. PATIENTS AND METHODS: A comparison of two groups of brain metastases treated with hypofractionated stereotactic radiotherapy or radiosurgery, with or without WBRT was performed. Hypofractionated stereotactic radiotherapy: there were eight series including 448 patients published from 2000 to 2009; treated with 5-6 MV X-Rays, non invasive head immobilization, a margin 2 to 10mm; 24 to 40Gy in three to five fractions; a 5 to 8 days duration in six series and 15-16 days in two other series. WBRT (30%) ; radiosurgery: there were 12 series (1994 to 2005) including 2157 patients; an invasive head immobilization, no margin; doses from 10 to 25 Gy; six series over 12 had Gamma Knife radiosurgery and six had Linacs X-Rays. WBRT (30 Gy/10 F/12 days) associated to radiosurgery in several series. The following parameters were compared: median GTV, median survival, 1-year survival rate, local control rate, necrosis and WBRT rates. RESULTS: Hypofractionated stereotactic radiotherapy series: the parameters were respectively: 0,52-4,47 cm(3) (median 2,8 cm(3)); 5 16 months (median 8,7 months); 68,2-93% (median 82,5%); necrosis rate 3,1%; associated WBRT 30%. Radiosurgery series: the parameters were respectively: 1,3 to 5,5 cm(3) (median 2 cm(3)); 5,5 to 22 months (median 11 months); 71 to 95% (median 85%); 0,5 to 6% (median 2,4%); associated WBRT 58%. Results seem similar in the two groups: Hypofractionated stereotactic radiotherapy with non invasive immobilization could theoretically treat all brain metastases sizes except lesions<10 mm (500 mm(3)). In large volumes,>4200 mm(3) GTV, the toxicity of hypofractionated stereotactic radiotherapy was not reported, thus it was difficult to compare its results with the published reports of radiosurgery toxicity. WBRT was a confusing parameter. Obviously, this initial survey has important limitations, specifically its methodology. CONCLUSION: Radiosurgery and hypofractionated stereotactic radiotherapy could be used to treat brain metastases with GTV>500 mm(3) and < or = 4200 mm(3) (O 20mm); for GTV<500 mm(3) (O 10mm) an invasive procedure with radiosurgery is necessary. For GTV>4200 mm(3) (O 20mm), hypofractionated stereotactic radiotherapy could be proposed, provided further studies, using 4 to 6 Gy fractions, a duration less or equal to 10-12 days and a margin of 2mm will be performed. PMID- 20004126 TI - Rationale for carbon ion therapy in high-grade glioma based on a review and a meta-analysis of neutron beam trials. AB - PURPOSE: The standard treatment of high-grade glioma is still unsatisfactory: the 2-year survival after radiotherapy being only 10-25%. A high linear energy transfer (LET) ionising radiotherapy has been used to overcome tumour radioresistance. An overview of the field is needed to justify future prospective controlled studies on carbon ion therapy. MATERIALS AND METHODS: A meta-analysis of clinical trials on neutron beam therapy and a literature review of clinical investigations on light ion use in high-grade glioma were carried out. RESULTS: Four randomised controlled trials on neutron beam therapy were retained. The meta analysis showed a non-significant 6% increase of two-year mortality (Relative risk [RR]=1.06 [0.97-1.15]) in comparison with photon therapy. Two phase I/II trials on carbon and neon ion therapy reported for glioblastoma 10% and 31% two year overall survivals and 13.9 and 19.0 months median survivals, respectively. CONCLUSION: This meta-analysis suggests that neutron beam therapy does not improve the survival of high-grade glioma patients while there is no definitive conclusion yet regarding carbon therapy. The ballistic accuracy and the improved biological efficacy of carbon ions renew the interest in prospective clinical trials on particle beam radiotherapy of glioma and let us expect favourable effects of dose escalation on patients' survival. PMID- 20004127 TI - Undiagnosed late-onset chylothorax accompanied by fatal acute pulmonary thromboembolism after surgical treatment of lung cancer: an autopsy case and review of the literature. AB - Late-onset chylothorax is a relatively rare and essentially benign postoperative complication. We describe an autopsy case of undiagnosed hemi-chylothorax accompanied by fatal acute pulmonary thromboembolism after pulmonary resection and lymph node dissection for lung cancer, reviewing the literature. The patient was a 60-year-old woman. She complained of dyspnea on the 20th postoperative day, which gradually increased. The patient suddenly died 2 days later. Autopsy demonstrated right chylothorax (1000 ml), and fatal acute pulmonary thromboembolism, mainly on the left side, originating from the lower extremities. Postoperative chylothorax, causing respiratory and pulmonary circulatory disturbance, may provide an important predisposition to ward fatality due to pulmonary thromboembolism even among those showing a milder type; thus careful management is necessary. PMID- 20004128 TI - Association of cardiac autonomic neuropathy with alteration of sympatho-vagal balance through heart rate variability analysis. AB - Early sub-clinical assessment of severity of cardiac autonomic neuropathy (CAN) and intervention are of prime importance for risk stratification and early treatment in preventing sudden death due to silent myocardial infarction. The Ewing battery is currently the diagnostic tool of choice but is unable to detect sub-clinical disease and requires patient cooperation. Time and frequency domain analysis have several shortcomings including sensitivity to recording length, respiratory activity and non-stationarities in the ECG signal. An important step forward is to have a non-invasive method of detecting CAN that is robust against these shortcomings and has a higher sensitivity for the presence of both sub clinical and overt clinical disease. This study presents a novel parameter, tone entropy (T-E) that analyses heart rate variability (HRV) of 20 min lead II ECG recordings. Tone (T) represents sympatho-vagal balance and entropy (E) the autonomic regularity activity. Thirteen normal subjects without (CAN-) and 21 with CAN (CAN+) participated in this study. Among 21 CAN+ subjects, 13 are early CAN+ (eCAN+), eight are definite CAN+ (dCAN+) according to autonomic nervous system function tests as described by Ewing. The results showed that tone was higher and the entropy was lower in the dCAN+ group (T=-0.033 to -0.010 and E=1.73-2.24) compared with the eCAN+ (T=-0.0927 to -0.0311 and E=2.0-2.65) and normal (T=-0.128 to -0.0635 and E=2.64-3.15) group. The research verified that T E is a suitable method to determine the presence of CAN that correctly identifies experimentally induced changes in cardiac function akin to parasympathetic and sympathetic dysfunction and differentiates between stages in CAN disease progression identified using the Ewing battery. PMID- 20004129 TI - Community off-sales provision and the presence of alcohol-related detritus in residential neighbourhoods. AB - This paper investigates the relationship between community off-sales premises and alcohol-related detritus (litter/remains) in residential neighbourhoods. This was accomplished by photographing all brand-identifiable alcohol product detritus (glass, packaging, etc.) where they lay and mapping these against the presence of off-sales outlets (licensed convenience stores) in the community. It was hypothesised that alcohol-related detritus would be greatest near to such alcohol outlets. However, although there was some evidence of a "broken bottles effect", accumulations of alcohol-related detritus near some off-sales premises, it is concluded that local area deprivation is a better predictor of such alcohol related incivility than is outlet provision. The implications of these findings are discussed in relation to current social responsibility policy developments which are designed to make the alcohol industry liable for alcohol-related incivilities. PMID- 20004130 TI - Objective versus subjective measures of the built environment, which are most effective in capturing associations with walking? AB - This study compared the strength of association of objective and subjective measures of environment with walking sufficiently for health. It used an existing model, which employed only objective environmental measures and adjusted for socio-demographic covariates, to estimate the influence of the built environment on walking. The original model was re-run with new, subjectively measured variables. The results showed that objective measures of the built environment had stronger associations with walking than subjective measures. Future studies of the environmental influences on physical activity should include objective measures of the built environment. PMID- 20004131 TI - The chemokine system in cancer biology and therapy. AB - Chemokines are a key component of cancer-related inflammation. Chemokines and chemokine receptors are downstream of genetic events that cause neoplastic transformation and are components of chronic inflammatory conditions, which predispose to cancer. Components of the chemokine system affect in a cell autonomous or non-autonomous way multiple pathways of tumor progression, including: leukocyte recruitment and function; cellular senescence; tumor cell proliferation and survival; invasion and metastasis. Available information in preclinical and clinical settings suggests that the chemokine system represents a valuable target for the development of innovative therapeutic strategies. PMID- 20004132 TI - Chromosomal numerical aberrations in apparently normal oral mucosa of heavy smokers affected by lung cancer. AB - Cigarette smoke creates a field of injury in the epithelial lining of the entire respiratory tract causing an increased risk for the development of malignant lesions. It is conceivable, therefore, that early genetic alterations, can be detected in oral mucosa of heavy smokers mainly those affected by lung cancer. As aneuploidy was shown to be an early event in oral carcinogenesis, we aimed to investigate the prevalence of aneuploid cells (ACs) in samples obtained from apparently normal looking oral mucosa of heavy smokers affected by lung cancer (LC). Two brush samples were collected from the oral mucosa of 152 subjects; 31 heavy smokers with LC, 59 heavy smokers without LC and 62 never-smokers. The samples were simultaneously analyzed for morphology and fluorescent in situ hybridization (FISH) using chromosomes 2 and 8 centromeric probes. Over 2% ACs were found in 23% of heavy smokers with LC compared to 12% in heavy smokers without LC and 5% of the never-smokers group (P=0.015). A trend was also noticed when comparing the group of heavy smokers without LC with the never-smokers (P=0.198). We conclude that heavy smokers harbour detectable chromosomal numerical aberrations in oral epithelial cells of normal looking mucosa. These aberrations are more frequently found in heavy smokers affected by LC. PMID- 20004133 TI - Biological pathways involved in the aggressive behavior of the keratocystic odontogenic tumor and possible implications for molecular oriented treatment - an overview. AB - In the classification of Head and Neck Tumors, published in 2005 by the World Health Organization Classification, the odontogenic keratocyst has been reclassified as a benign intraosseous neoplasm, calling it "keratocystic odontogenic tumor" (KCOT). Significant differences on the molecular level between KCOT and other odontogenic cystic lesions suggest a different biological origin. Genetic and molecular research regarding odontogenic tumors, and KCOTs in particular, has led to an increasing amount of knowledge and understanding of their physiopathological pathways. A review of the biological behavior of this recognized aggressive pathological entity of the jaws and a contemporary outline of the molecular (growth factors, p53, PCNA and Ki-67, bcl-2) and genetic (PTCH, SHH) alterations associated with this odontogenic neoplasm provides a better understanding of the mechanisms involved in its development and strengthen the current concept that the KCOT should, indeed, be regarded as a neoplasm. Furthermore, markers known to be rapidly induced in response to growth factors, tumor promoters, cytokines, bacterial endotoxins, oncogenes, hormones and shear stress, such as COX-2, may also shed new light on the biological mechanisms involved in the development of these benign but sometimes aggressive neoplasms of the jaws. PMID- 20004134 TI - Ameloblastoma of the jaws: a critical reappraisal based on a 40-years single institution experience. AB - The 40-years of experience in a single institution with the treatment of previously untreated ameloblastoma have been reported, followed by a management protocol. Retrospectively, 25 consecutive patients treated between 1969 and 2009 have been analyzed. In 11 patients, a preoperative diagnosis of ameloblastoma was available. In the remaining 14 patients the diagnosis of ameloblastoma was a postoperative one. For the recurrence rate a minimum follow-up period of 5 years was observed; 20 patients met these criteria. After primary radical surgery in five patients, no recurrences were observed. In case of conservative surgical treatment, performed in 15 patients, a recurrence was observed in eight (53%) of them. Six of these patients were then treated successfully by radical surgery, while two patients refused such surgical approach. In one of the patients with a recurrence a cervical lymph node metastasis was detected at the same time, resulting in a diagnosis of metastasizing ameloblastoma. PMID- 20004135 TI - Employment and return to work in head and neck cancer survivors. AB - To investigate employment in working-age head and neck cancer (HNC) survivors before and after treatment in relation to sociodemographic and clinical factors, health related quality of life, and emotional distress. Patients younger than 65 years at time of diagnosis and at least 2 years after curative treatment for HNC were included. Exclusion criteria were cognitive dysfunction and no understanding of Dutch language. Primary outcome measures were employment status and return to work assessed by a study specific questionnaire. Secondary outcome measures were health related quality of life (EORTC QLQ-C30 and QLQ-H&N35) and emotional distress (Hospital Anxiety and Depression Scale (HADS)). Sociodemographic and disease and treatment related parameters, health related quality of life, and distress were compared to employment status. Eighty-five out of 113 included patients completed the questionnaires (response rate 75%). At time of diagnosis 32 patients were not working (38%), significantly associated with (higher) age and (lower) education level. Of the 53 patients who were employed at time of diagnosis, 44 patients returned to work (83%): 28 to the same work, 7 to adapted work and 9 to other work. Median time was 6 months to return to work (range 0-24 months) and 71% of the patient returned to work within 6 months after treatment. Anxiety and oral dysfunction as xerostomia, trismus, sticky saliva, problems with teeth, and loss of appetite, problems with social eating and social contacts were significantly associated with employment after treatment. The majority of employed HNC survivors return to work within 6 months after treatment. Oral dysfunction, loss of appetite, deteriorated social functioning, and high levels of anxiety are barriers for HNC survivors to return to work after treatment. PMID- 20004136 TI - Proteomics shed light on the brassinosteroid signaling mechanisms. AB - Large numbers of receptor-like kinases (RLKs) play key roles in plant development and defense by perceiving extracellular signals. The mechanisms of ligand-induced kinase activation and downstream signal transduction have been studied for only a few RLK pathways, among which the brassinosteroid (BR) pathway is the best characterized. Recently, proteomics studies identified new components that bridge the last gap in the genetically defined BR-signaling pathway, establishing the first complete pathway from an RLK to transcription factors in plants. Furthermore, analyses of phosphorylation events, mostly by mass spectrometry, provided insights into the mechanistic details of receptor kinase activation and regulation of downstream components by phosphorylation. This review focuses on recent progress in understanding BR signal transduction made by proteomics studies. PMID- 20004137 TI - The manganese(III) complex with chelating Schiff base ligand: X-ray structure, spectroscopic and computational studies. AB - A new Mn(III) complex, [MnCl(H(2)O)(L)].H(2)O.C(2)H(5)OH, where L=2,2'-[1,2 phenylenebis[nitrilomethylylidene]]bis(6-methoxyphenolate), has been synthesized and characterized by single-crystal X-ray diffraction. There is a good agreement between calculated and experimental structural data. The complex is crystallized in orthorhombic with space group Pbca. The Mn1 atom is coordinated with one Schiff base ligand, one water molecule and one chloride anion, forming a six coordination number. The electronic and fluorescence spectra of the complex were also studied. PMID- 20004138 TI - Multi-wavelength spectrophotometric determination of acidity constant of some newly synthesized Schiff bases and their QSPR study. AB - The acidity constants of some newly synthesized Schiff base derivatives were determined by hard-model based multivariate data analysis of the spectrophotometric data in the course of pH-metric titration in 50% (v/v) methanol-water binary solvent. The employed data analysis method was also able to extract the pure spectra and pH-dependent concentration profiles of the acid-base species. The molecules that possess different substituents (both electron donating and withdrawing) on the ortho-, meta- and para-positions of one of the phenyl ring showed variable acidity constants ranging from 8.77 to 11.07 whereas the parent molecule had an acidity constant of 10.25. To investigate the quantitative effects of changing of substitution pattern on the acidity constant, a quantitative structure-property relation analysis was conducted using substituent constants and molecular descriptor. Some models with high statistical quality (measured by cross-validation Q(2)) were obtained. It was found that the acidity constant of the studied molecules in the methanol-water mixed solvent not only is affected by electronic features of the solutes but also by the lipophilic interaction between methanol part of solvent and the deprotonated solutes. PMID- 20004139 TI - Synthesis, spectroscopic, magnetic and thermal properties of bimetallic salts, [Ni(L)][MCl4] [where M=Co(II), Zn(II), Hg(II) and L=3,7-bis(2-aminoethyl)-1,3,5,7 tetraazabicyclo(3.3.1)nonane]. X-ray structure of [Ni(L)][CoCl4]. AB - New bimetallic complex salts corresponding to the formulation [Ni(L)][MCl(4)] have been synthesized by the facile reaction between [Ni(L)](ClO(4))(2) and [MCl(2)(PPh(3))(2)] in high yields [where M=Co(II), Zn(II), Hg(II) and L=3,7 bis(2-aminoethyl)-1,3,5,7-tetraazabicyclo(3.3.1)nonane]. The complexes were characterized by IR, electronic spectra, TGA/DSC, magnetic moment and conductivity measurements. The X-ray crystal structure for [Ni(L)][CoCl(4)] clearly establishes the cationic-anionic interaction. It crystallizes in the space group P1 with unit cell dimensions a=7.1740(15)A, b=8.1583(16)A and c=8.3102(16)A. A square-planar geometry is evident for the [Ni(L)](2+) cation while the anion is found to be tetrahedral. A two-step thermolytic pattern is observed in the pyrolysis of the bimetallic complex salts. PMID- 20004140 TI - Absorption and fluorescence properties of aryl substituted porphyrins in different media. AB - Absorption and fluorescence properties of aryl substituted porphyrins, 5,10,15,20 tetra-4-oxy(aceticacid)phenylporphyrin (TAPP), 5,10,15,20-tetra-(4-phenoxyphenyl) porphyrin (TPPP), 5,10,15,20-tetra-(3-bromo-4-hydroxyphenyl) porphyrin (TBHPP), and 5,10,15,20-tetra-p-chloromethylphenyl porphyrin (CMPP) were investigated. The UV/vis absorption, fluorescence and excited spectra as the fluorescence quantum yields and fluorescence lifetimes for the compounds were measured in organic solvents (chloroform (CHCl(3)), tetrahydrofuran (THF)) and immobilized media (PVC film, sol-gel matrix). The fluorescence quantum yields of TAPP and TPPP were higher than the others. The fluorescence lifetimes of all studied porphyrin derivates were found to be fifty percent lower and their fluorescence intensities were increased fifty percent more in both of immobilized mediums, as compared to organic solvents. PMID- 20004141 TI - The effect of nitric acid (HNO3) on growth, spectral, thermal and dielectric properties of triglycine sulphate (TGS) crystal. AB - The effect of nitric acid (HNO(3)) addition on the growth of triglycine sulphate (TGS) crystal has been studied from the aqueous solution for various concentrations of nitric acid. Significant changes in the crystal size and morphology have been observed in all the grown samples. Single crystal and powder X-ray diffraction analyses confirm the structure and cell parameter values of pure and HNO(3) doped TGS crystals. FT-Raman and FTIR spectra confirm the characteristics absorption bands of pure and HNO(3) doped TGS crystals. The composition of TGS crystals have been confirmed by CHNS analysis. Physical properties such as thermal, dielectric and mechanical studies have been performed for the pure and HNO(3) doped TGS crystals. The dielectric constants of the crystals have been studied as a function of frequency. The results suggest that the HNO(3) is doped into TGS crystal and that the doping increases its dielectric constant. PMID- 20004142 TI - Synthesis, crystal structure and photoluminescence of 3-(4-(anthracen-10 yl)phenyl)-benzo[5,6]coumarin. AB - A new coumarin derivative, 3-(4-(anthracen-10-yl)phenyl)-benzo[5,6]coumarin, was synthesized and characterized by FT-IR, (1)H NMR, element analysis and single crystal X-ray crystallography. The dihedral angle of benzo[5,6]coumarin ring and phenyl group is 36.15 degrees, and the dihedral angle of phenyl group and anthracene skeleton is 89.37 degrees. The UV-vis absorption and photoluminescence of the compound were discussed. The result shown that the compound exhibits high fluorescence quantum yield (Phi(F)), large Stokes shift and green emission (508 nm). The molecular structure of the compound was optimized using density functional theory (DFT) at B3LYP/6-31G(d) level, and the HOMO and LUMO levels of the compound were deduced. PMID- 20004143 TI - Unsupervised self-initiation of antiretroviral drugs in a newly diagnosed HIV-1 infected haemodialysis patient. PMID- 20004144 TI - Febrile illness in a returned traveller from Thailand. PMID- 20004145 TI - Factors associated with persistence of arthralgia among Chikungunya virus infected travellers: report of 42 French cases. AB - BACKGROUND: In 2005-2006, a major epidemic of CHIKV infection occurred in the Islands of the south-western Indian Ocean, and longstanding manifestations seemed to be more frequent than described before. OBJECTIVES: To describe the frequency and related factors of late clinical manifestations of CHIKV infection among imported cases living in Aquitaine area, France. STUDY DESIGN: All patients recruited through the travel clinic and tropical medicine unit of the University Hospital Centre of Bordeaux with possible CHIKV infection were prospectively recorded, and confirmed cases of CHIKV infection were interviewed 2 years after infection. Factors associated with the persistence of symptoms were determined by multivariate logistic regression. RESULTS: Among the 29 cases followed, 17 still suffered from arthralgia 2 years after infection, and most of them had never recovered from the initial phase of the condition. The risk of persistent arthralgia tended to be higher among subjects with low educational level, subjects infected in the Reunion Island, and when initial phase lasted 30 days or more and was characterised by a severe pain. CONCLUSIONS: Consistent with previous studies, our findings showed worsened late manifestations among patients returning from Indian Ocean area. Persistence of symptoms tended to be linked with clinical burden during the acute phase, which can be informative for early recognition and management of patients at risk for developing persistent rheumatic symptoms. Cryoglobulins failed to be identified in seronegative patients with invalidating dengue-like syndrome. PMID- 20004146 TI - Viral load and genotypes of noroviruses in symptomatic and asymptomatic children in Southeastern Brazil. AB - BACKGROUND: Noroviruses (NoVs) are a major etiological agent of sporadic acute gastroenteritis worldwide. OBJECTIVES: To detect, quantify and characterize genogroups and genotypes of NoVs in children with and without gastrointestinal symptoms. STUDY DESIGN: NoVs were investigated by RT-PCR in a total of 319 fecal specimens from children up to three years old with (n=229) and without (n=90) acute diarrhea, between February 2003 and June 2004 in the emergency room in Vitoria, Southeastern Brazil. NoVs were quantified by real-time PCR and genotyped. RESULTS: NoVs were detected in 17% (40/229) and 13% (12/90) of symptomatic and asymptomatic children, respectively. Six NoV-rotavirus A mixed infections were observed. Fifty-one strains were characterized as NoV GII and one as GI. Twenty strains were characterized as GII/4 (9/13), GII/3 (1/13), GII/6 (2/13) and GII/14 (1/13) in symptomatic and GII/3 (6/7) and GII/8 (1/7) in asymptomatic children. The median RNA viral loads were 8.39 and 7.15log(10)copies/g of fecal specimens for symptomatic and asymptomatic children, respectively (p=0.011). NoV load was lower when it was present in a mixed infection with rotavirus A (p=0.0005). CONCLUSIONS: This study demonstrates a diversity of NoV strains circulating in this geographic area, and reports GII/8 and GII/14 in the American Continent for the first time. In addition, it confirms GII/4 as the most prevalent genotype in symptomatic children and identified GII/3 in an important frequency, especially in asymptomatic children. Furthermore, preliminary results show that symptomatic patients present a viral load that is significantly greater than asymptomatic children (p=0.011). PMID- 20004147 TI - Control of central self-tolerance induction by autoreactive CD4+ thymocytes. AB - Medullary thymic epithelial cells (mTECs) are crucial for the selection of a T cell-receptor (TCR) repertoire purged of self-reactive specificities, because these cells activate a promiscuous gene-expression program that leads to the synthesis of a wide array of peripheral tissue-restricted self-antigens. This review summarizes recent progress in our understanding of the cellular interactions, ligands, receptors and signal-transduction pathways that control mature-mTEC development. The particular focus is on new findings supporting the model that mature-mTEC development in the postnatal thymus depends on nuclear factor-kappaB (NF-kappaB) signaling induced by CD40-CD40 ligand, and receptor activator-of-NF-kappaB (RANK)-RANK ligand interactions, and that these signals are delivered in the context of antigen-specific interactions between CD4(+) thymocytes carrying autoreactive TCRs and mTECs displaying cognate autoantigen MHC-class-II complexes. PMID- 20004148 TI - Evaluation of sleep quality in patients with refractory seizures who undergo epilepsy surgery. AB - The aim of the study was to evaluate excessive daytime sleepiness and subjective sleep quality in patients who undergo epilepsy surgery for treatment of refractory partial seizures. Forty-eight patients were enrolled in this research study. All of them were evaluated 2 days before and 3 months after the surgery. Two questionnaires were used to assess daytime sleepiness (Epworth Sleepiness Scale [ESS]) and sleep quality (Pittsburgh Sleep Quality Index [PSQI]). Global PSQI was high (mean=5.65 SD=3.71) before the surgical procedure (P<0.001). PSQI evaluation revealed higher and statistically significant scores in three components as well as in the global score, when analyzed by predominance of daytime or nocturnal seizures. ESS and PSQI scores were also analyzed by gender, antiepileptic drug class, age, and seizure frequency, with no significant differences. We concluded that patients with partial recurrent seizures of temporal origin have poor subjective sleep quality that improves significantly after epilepsy surgery. PMID- 20004150 TI - Genotype based evaluation of Pseudomonas aeruginosa eradication treatment success in cystic fibrosis patients. AB - BACKGROUND: Longitudinal data regarding the genotypes of Pseudomonas aeruginosa isolates after eradication treatment are limited. We followed cystic fibrosis patients after a first ever isolation of P. aeruginosa and evaluated the P. aeruginosa-free time period after eradication therapy. METHODS: Between January 2003 and December 2008 respiratory samples were cultured prospectively from 41 patients with a first ever P. aeruginosa isolate. Twenty five patients had at least one subsequent isolate. Treatment efficacy was assessed based on the time to a second isolation and on comparison of the RAPD genotypes of the P. aeruginosa isolates. RESULTS: Eleven patients became chronically colonized during the study period. For ten of these the second isolate had the same genotype as the first isolate. Moreover, these patients had a significantly shorter P. aeruginosa-free time interval between the first ever and the second isolate compared to the 14 not chronically colonized patients (median 0 months versus 7.5 months, p<0.05). CONCLUSION: Our results indicate that the presence of a genotypically identical subsequent P. aeruginosa isolate and/or a short P. aeruginosa-free time interval after treatment are ominous signs and might be useful additional tools to predict impending chronic colonization. Current routine bacteriological methods for the detection of P. aeruginosa may lack the sensitivity to discriminate between true eradication and low bacterial persistence. PMID- 20004151 TI - Constipation in pediatric cystic fibrosis patients: an underestimated medical condition. AB - BACKGROUND: The aims of this study were to determine prevalence, risk factors and treatment of constipation in patients with Cystic Fibrosis (CF), as well as the diagnostic value of abdominal radiography. METHODS: A cohort of 214 pediatric CF patients was investigated. Furthermore, 106 abdominal radiographs of CF patients with or without constipation were independently assessed by three observers on two separate occasions using the Barr and Leech scores. RESULTS: The prevalence of constipation was 47%. Low total fat absorption and meconium ileus were independent risk factors for constipation in CF, while fiber and fluid intake were not associated. In CF patients the inter and intraobserver variabilities of the Barr and Leech scores were poor to moderate. CONCLUSION: Constipation is a significant medical issue in CF and was associated with low total fat absorption and a history of meconium ileus. Finally, abdominal radiography seems of little value in the regular follow-up of CF patients. PMID- 20004149 TI - Nuclear reorganization of DNA mismatch repair proteins in response to DNA damage. AB - The DNA mismatch repair (MMR) system is highly conserved and vital for preserving genomic integrity. Current mechanistic models for MMR are mainly derived from in vitro assays including reconstitution of strand-specific MMR and DNA binding assays using short oligonucleotides. However, fundamental questions regarding the mechanism and regulation in the context of cellular DNA replication remain. Using synchronized populations of HeLa cells we demonstrated that hMSH2, hMLH1 and PCNA localize to the chromatin during S-phase, and accumulate to a greater extent in cells treated with a DNA alkylating agent. In addition, using small interfering RNA to deplete hMSH2, we demonstrated that hMLH1 localization to the chromatin is hMSH2-dependent. hMSH2/hMLH1/PCNA proteins, when associated with the chromatin, form a complex that is greatly enhanced by DNA damage. The DNA damage caused by high doses of alkylating agents leads to a G(2) arrest after only one round of replication. In these G(2)-arrested cells, an hMSH2/hMLH1 complex persists on chromatin, however, PCNA is no longer in the complex. Cells treated with a lower dose of alkylating agent require two rounds of replication before cells arrest in G(2). In the first S-phase, the MMR proteins form a complex with PCNA, however, during the second S-phase PCNA is missing from that complex. The distinction between these complexes may suggest separate functions for the MMR proteins in damage repair and signaling. Additionally, using confocal immunofluorescence, we observed a population of hMSH6 that localized to the nucleolus. This population is significantly reduced after DNA damage suggesting that the protein is shuttled out of the nucleolus in response to damage. In contrast, hMLH1 is excluded from the nucleolus at all times. Thus, the nucleolus may act to segregate a population of hMSH2-hMSH6 from hMLH1-hPMS2 such that, in the absence of DNA damage, an inappropriate response is not invoked. PMID- 20004152 TI - Garlic extract prevents CCl(4)-induced liver fibrosis in rats: The role of tissue transglutaminase. AB - BACKGROUND AND AIM: Tissue transglutaminase contributes to liver damage in the development of hepatic fibrosis. In a model of neurodegeneration, the therapeutic benefit of cystamine has been partly attributed to its inhibition of transglutaminase activity. Garlic extract contains many compounds structurally related to cystamine. We investigated the anti-fibrotic effect of garlic extract and cystamine as specific tissue transglutaminase inhibitors. METHODS: Rat liver fibrosis was induced by intraperitoneal injection of carbon tetrachloride (CCl(4)) for 7 weeks. Cystamine or garlic extract was administrated by daily intraperitoneal injection, starting from the day after the first administration of CCl(4). Hepatic function, histology, tissue transglutaminase immunostaining and image analysis to quantify Red Sirius stained collagen deposition were examined. Reverse transcription-polymerase chain reaction to detect alpha-SMA, IL 1beta and tissue transglutaminase expression and Western blot for tissue transglutaminase protein amount were performed. Transglutaminase activity was assayed on liver homogenates by a radio-enzymatic method. RESULTS: Transglutaminase activity was increased in CCl(4) group and reduced by cystamine and garlic extract (p<0.05). Treatment with cystamine and garlic extract reduced the liver fibrosis and collagen deposition, particularly in the garlic extract group (p<0.01). Moreover, the liver damage improved and serum alanine aminotransferase was decreased (p<0.05). Tissue transglutaminase immunolocalised with collagen fibres and is mainly found in the ECM of damaged liver. Alpha-SMA, IL-1beta, tissue transglutaminase mRNA and tissue transglutaminase protein were down-regulated in the cystamine and garlic extract groups compared to controls. CONCLUSION: These findings concurrently suggest that transglutaminase may play a pivotal role in the pathogenesis of liver fibrosis and may identify garlic cystamine-like molecules as a potential therapeutic strategy in the treatment of liver injury. PMID- 20004153 TI - [Bronchopulmonary dysplasia and prematurity. Short-and long-term respiratory changes]. AB - Bronchopulmonary dysplasia (BPD) is the most frequent chronic lung disease in premature children. With the inclusion of antenatal steroid therapy, surfactant use and novel mechanical ventilation strategies, survival of premature newborns has increased, whereupon the incidence of BPD has not only decreased but has also risen in extremely premature newborns. This has led to a high respiratory morbidity in the first 2-3 years of life, with numerous admissions to hospital and respiratory exacerbations mostly due to viral infections. Although there is a trend towards improvement, during school age and adolescence, respiratory symptoms may persist, due to changes in pulmonary function often showing a lower exercise capacity. Although BPD symptoms are similar to those of asthma, as there is limitation in airflow and bronchial hyperresponsiveness (BHR), pathophysiological mechanisms could be different in both diseases. On the other hand, isolated prematurity plays an important role in the child's respiratory pathology, proving that pulmonary function alterations in preterm children are present since the first months of life. A higher respiratory morbidity has also been observed in these children when compared to full-term newborns, not only during the first years of life but also subsequently. In this study, different aspects of chronic respiratory disease associated with prematurity will be analysed, drawing special attention to clinical symptoms, respiratory function changes, BHR and exercise capacity. All these aspects will be reviewed from early childhood until adolescence and young adult age. Similarities and differences between BPD and asthma will also be discussed. PMID- 20004155 TI - Sensitivity to change of the Quebec Back Pain Disability Scale and the Dallas Pain Questionnaire. AB - OBJECTIVE: To study and compare the sensitivities to change of the French versions of the Quebec Back Pain Disability Scale (QBPDS) and the Dallas Pain Questionnaire (DPQ). METHOD: A retrospective study of the medical records of 30 chronic low back pain (LBP) outpatients undergoing a 4-week functional training program in a physical medicine and rehabilitation unit. Physical data (a visual analogue pain scale, the Schober index, Sorensen and Shirado tests and the finger to-floor distance) and functional status (QBPDS and DPQ scores) were measured at D0 and D25. Sensitivity to change was assessed in terms of the standardized response mean (SRM) and the effect size (ES). RESULTS: With the exception of the DPQ's "social interest" domain, all QBPDS and DPQ scores improved significantly over the course of the rehabilitation programme. Sensitivity to change was high for the QBPDS (SRM=0.80; ES=0.62) but only moderate for the DPQ (SRM=0.59; ES=0.61). DISCUSSION-CONCLUSION: Our results show that the QBPDS was more sensitive to clinical change than the DPQ in a sample of chronic LBP sufferers. It would be interesting to assess this sensitivity in a population of acute LBP patients and to evaluate the long-term effects of a functional rehabilitation program on the DPQ's "social interest" domain. PMID- 20004154 TI - HMGNs, DNA repair and cancer. AB - DNA lesions threaten the integrity of the genome and are a major factor in cancer formation and progression. Eukaryotic DNA is organized in nucleosome-based higher order structures, which form the chromatin fiber. In recent years, considerable knowledge has been gained on the importance of chromatin dynamics for the cellular response to DNA damage and for the ability to repair DNA lesions. High Mobility Group N1 (HMGN1) protein is an emerging factor that is important for chromatin alterations in response to DNA damage originated from both ultra violet light (UV) and ionizing irradiation (IR). HMGN1 is a member in the HMGN family of chromatin architectural proteins. HMGNs bind directly to nucleosomes and modulate the structure of the chromatin fiber in a highly dynamic manner. This review focuses mainly on the roles of HMGN1 in the cellular response pathways to different types of DNA lesions and in transcriptional regulation of cancer related genes. In addition, emerging roles for HMGN5 in cancer progression and for HMGN2 as a potential tool in cancer therapy will be discussed. PMID- 20004156 TI - Expanding the ribosomal universe. AB - In this issue of Structure, Taylor et al. (2009) present the most complete model of an eukaryotic ribosome to date. This achievement represents a critical milestone along the path to structurally defining the unique aspects of the eukaryotic protein synthetic machinery. PMID- 20004157 TI - Insights into resistance against lincosamide antibiotics. AB - Bacteria utilize multiple strategies to circumvent antibiotics, producing broad specificity exporters or enzymes that catalyze the modification of either antibiotics or their targets. A report in this issue of Structure provides the structural and catalytic mechanisms of LinB, an adenylyltransferase of E. faecium that confers resistance to the lincosamide antibiotic clindamycin. PMID- 20004159 TI - E pluribus tres: the 2009 nobel prize in chemistry. AB - This year's Nobel Prize in Chemistry celebrates a multitude of research areas, making the difficult selection of those most responsible for providing atomic details of the nanomachine that makes proteins according to genetic instructions. The Ribosome and RNA polymerase (recognized in 2006) structures highlight a puzzling asymmetry at the origins of biology. PMID- 20004158 TI - Cyclic nucleotide binding GAF domains from phosphodiesterases: structural and mechanistic insights. AB - GAF domains regulate the catalytic activity of certain vertebrate cyclic nucleotide phosphodiesterases (PDEs) by allosteric, noncatalytic binding of cyclic nucleotides. GAF domains arranged in tandem are found in PDE2, -5, -6, 10, and -11, all of which regulate the cellular concentrations of the second messengers cAMP and/or cGMP. Nucleotide binding to GAF domains affects the overall conformation and the catalytic activity of full-length PDEs. The cyclic nucleotide-bound GAF domains from PDE2, -5, -6, and -10 all adopt a conserved fold but show subtle differences within the binding pocket architecture that account for a large range of nucleotide affinities and selectivity. NMR data and details from the structure of full-length nucleotide-free PDE2A reveal the dynamic nature and magnitude of the conformational change that accompanies nucleotide binding. The discussed GAF domain structures further reveal differences in dimerization properties and highlight the structural diversity within GAF domain-containing PDEs. PMID- 20004160 TI - Averaging of electron subtomograms and random conical tilt reconstructions through likelihood optimization. AB - The reference-free averaging of three-dimensional electron microscopy (3D-EM) reconstructions with empty regions in Fourier space represents a pressing problem in electron tomography and single-particle analysis. We present a maximum likelihood algorithm for the simultaneous alignment and classification of subtomograms or random conical tilt (RCT) reconstructions, where the Fourier components in the missing data regions are treated as hidden variables. The behavior of this algorithm was explored using tests on simulated data, while application to experimental data was shown to yield unsupervised class averages for subtomograms of groEL/groES complexes and RCT reconstructions of p53. The latter application served to obtain a reliable de novo structure for p53 that may resolve uncertainties about its quaternary structure. PMID- 20004161 TI - Crystallographic insight into collagen recognition by discoidin domain receptor 2. AB - The discoidin domain receptors, DDR1 and DDR2, are widely expressed receptor tyrosine kinases that are activated by triple-helical collagen. They control important aspects of cell behavior and are dysregulated in several human diseases. The major DDR2-binding site in collagens I-III is a GVMGFO motif (O is hydroxyproline) that also binds the matricellular protein SPARC. We have determined the crystal structure of the discoidin domain of human DDR2 bound to a triple-helical collagen peptide. The GVMGFO motifs of two collagen chains are recognized by an amphiphilic pocket delimited by a functionally critical tryptophan residue and a buried salt bridge. Collagen binding results in structural changes of DDR2 surface loops that may be linked to the process of receptor activation. A comparison of the GVMGFO-binding sites of DDR2 and SPARC reveals a striking case of convergent evolution in collagen recognition. PMID- 20004162 TI - Non-native interactions are critical for mechanical strength in PKD domains. AB - Experimental observation has led to the commonly held view that native state protein topology is the principle determinant of mechanical strength. However, the PKD domains of polycystin-1 challenge this assumption: they are stronger than predicted from their native structure. Molecular dynamics simulations suggest that force induces rearrangement to an intermediate structure, with nonnative hydrogen bonds, that resists unfolding. Here we test this hypothesis directly by introducing mutations designed to prevent formation of these nonnative interactions. We find that these mutations, which only moderately destabilize the native state, reduce the mechanical stability dramatically. The results demonstrate that nonnative interactions impart significant mechanical stability, necessary for the mechanosensor function of polycystin-1. Remarkably, such nonnative interactions result from force-induced conformational change: the PKD domain is strengthened by the application of force. PMID- 20004164 TI - Global conformational changes of ribosome observed by normal mode fitting for 3D Cryo-EM structures. AB - Many three-dimensional density maps of 70S ribosome at various functional states are available now in the Electron Microscopy DataBank at EBI. We used our new flexible-fitting approach to systematically analyze these maps to reveal the global conformational differences between the EM structures. Large-scale ratchet like deformations were observed in an EM structure of the initiation complex and in some EM structures bound with EFG, RF3, and RRF. In most of them, the L1 stalk, which interacts with the tRNA molecule at the E site of ribosome and is considered to be involved in the release of the tRNA, was in "the blocking state" for the E-tRNA. Furthermore, we found that the EM structures bound with EFG or RRF were aligned in the conformational space, suggesting that the large-scale conformational changes of the 70S ribosome bound with these factors occur along a specific pathway in a concerted manner. PMID- 20004165 TI - Unraveling the allosteric mechanism of serine protease inhibition by an antibody. AB - Recent structural studies have outlined the mechanism of protease inhibition by active site-directed antibodies. However, the molecular basis of allosteric inhibition by antibodies has been elusive. Here we report the 2.35 A resolution structure of the trypsin-like serine protease hepatocyte growth factor activator (HGFA) in complex with the allosteric antibody Ab40, a potent inhibitor of HGFA catalytic activity. The antibody binds at the periphery of the substrate binding cleft and imposes a conformational change on the entire 99-loop (chymotrypsinogen numbering). The altered conformation of the 99-loop is incompatible with substrate binding due to the partial collapse of subsite S2 and the reorganization of subsite S4. Remarkably, a single residue deletion of Ab40 abolished inhibition of HGFA activity, commensurate with the reversal of the 99 loop conformation to its "competent" state. The results define an "allosteric switch" mechanism as the basis of protease inhibition by an allosteric antibody. PMID- 20004163 TI - Comprehensive molecular structure of the eukaryotic ribosome. AB - Despite the emergence of a large number of X-ray crystallographic models of the bacterial 70S ribosome over the past decade, an accurate atomic model of the eukaryotic 80S ribosome is still not available. Eukaryotic ribosomes possess more ribosomal proteins and ribosomal RNA than do bacterial ribosomes, which are implicated in extraribosomal functions in the eukaryotic cells. By combining cryo EM with RNA and protein homology modeling, we obtained an atomic model of the yeast 80S ribosome complete with all ribosomal RNA expansion segments and all ribosomal proteins for which a structural homolog can be identified. Mutation or deletion of 80S ribosomal proteins can abrogate maturation of the ribosome, leading to several human diseases. We have localized one such protein unique to eukaryotes, rpS19e, whose mutations are associated with Diamond-Blackfan anemia in humans. Additionally, we characterize crucial interactions between the dynamic stalk base of the ribosome with eukaryotic elongation factor 2. PMID- 20004166 TI - Structure of HIV-1 reverse transcriptase with the inhibitor beta-Thujaplicinol bound at the RNase H active site. AB - Novel inhibitors are needed to counteract the rapid emergence of drug-resistant HIV variants. HIV-1 reverse transcriptase (RT) has both DNA polymerase and RNase H (RNH) enzymatic activities, but approved drugs that inhibit RT target the polymerase. Inhibitors that act against new targets, such as RNH, should be effective against all of the current drug-resistant variants. Here, we present 2.80 A and 2.04 A resolution crystal structures of an RNH inhibitor, beta thujaplicinol, bound at the RNH active site of both HIV-1 RT and an isolated RNH domain. beta-thujaplicinol chelates two divalent metal ions at the RNH active site. We provide biochemical evidence that beta-thujaplicinol is a slow-binding RNH inhibitor with noncompetitive kinetics and suggest that it forms a tropylium ion that interacts favorably with RT and the RNA:DNA substrate. PMID- 20004167 TI - Identification of structural mechanisms of HIV-1 protease specificity using computational peptide docking: implications for drug resistance. AB - Drug-resistant mutations (DRMs) in HIV-1 protease are a major challenge to antiretroviral therapy. Protease-substrate interactions that are determined to be critical for native selectivity could serve as robust targets for drug design that are immune to DRMs. In order to identify the structural mechanisms of selectivity, we developed a peptide-docking algorithm to predict the atomic structure of protease-substrate complexes and applied it to a large and diverse set of cleavable and noncleavable peptides. Cleavable peptides showed significantly lower energies of interaction than noncleavable peptides with six protease active-site residues playing the most significant role in discrimination. Surprisingly, all six residues correspond to sequence positions associated with drug resistance mutations, demonstrating that the very residues that are responsible for native substrate specificity in HIV-1 protease are altered during its evolution to drug resistance, suggesting that drug resistance and substrate selectivity may share common mechanisms. PMID- 20004168 TI - Structure and mechanism of the lincosamide antibiotic adenylyltransferase LinB. AB - Lincosamides make up an important class of antibiotics used against a wide range of pathogens, including methicillin-resistant Staphylococcus aureus. Predictably, lincosamide-resistant microorganisms have emerged with antibiotic modification as one of their major resistance strategies. Inactivating enzymes LinB/A catalyze adenylylation of the drug; however, little is known about their mechanistic and structural properties. We determined two X-ray structures of LinB: ternary substrate- and binary product-bound complexes. Structural and kinetic characterization of LinB, mutagenesis, solvent isotope effect, and product inhibition studies are consistent with a mechanism involving direct in-line nucleotidyl transfer. The characterization of LinB enabled its classification as a member of a nucleotidyltransferase superfamily, along with nucleotide polymerases and aminoglycoside nucleotidyltransferases, and this relationship offers further support for the LinB mechanism. The LinB structure provides an evolutionary link to ancient nucleotide polymerases and suggests that, like protein kinases and acetyltransferases, these are proto-resistance elements from which drug resistance can evolve. PMID- 20004169 TI - A role for a specific cholesterol interaction in stabilizing the Apo configuration of the human A(2A) adenosine receptor. AB - The function of G-protein-coupled receptors is tightly modulated by the lipid environment. Long-timescale molecular dynamics simulations (totaling approximately 3 mus) of the A(2A) receptor in cholesterol-free bilayers, with and without the antagonist ZM241385 bound, demonstrate the instability of helix II in the apo receptor in cholesterol-poor membrane regions. We directly observe that the effect of cholesterol binding is to stabilize helix II against a buckling type deformation, perhaps rationalizing the observation that the A(2A) receptor couples to G protein only in the presence of cholesterol (Zezula and Freissmuth, 2008). The results suggest a mechanism by which the A(2A) receptor may function as a coincidence detector, activating only in the presence of both cholesterol and agonist. We also observed a previously hypothesized conformation of the tryptophan "rotameric switch" on helix VI in which a phenylalanine on helix V positions the tryptophan out of the ligand binding pocket. PMID- 20004170 TI - Structural basis of the cofactor- and substrate-assisted activation of human coagulation factor IXa. AB - Human coagulation factor IX serves both to maintain and to control blood coagulation. The dual function of this hemophilic factor is implemented by a tiered activation mechanism. Processed two-chain factor IXa is catalytically silent; only together with its cofactor VIIIa does factor IXa form the highly potent Xase complex. The detailed mechanism of this secondary activation has remained elusive so far. Here we present the crystal structures of Xase-like factor IXa mutants with several-thousand-fold activity enhancement that mimic the secondary activation by Xase formation. The structures reveal how cofactor triggered and substrate-assisted modulations in the factor IXa 99- and 60-loops cooperate in S4 through S2' formation, allowing for productive substrate recognition. We could further physically map and visualize a distinct communication line, along which agonists such as Ca(2+) direct their effects to the active site and vice versa. PMID- 20004172 TI - Structure-function studies of chemokine-derived carboxy-terminal antimicrobial peptides. AB - Recent reports which show that several chemokines can act as direct microbicidal agents have drawn renewed attention to these chemotactic signalling proteins. Here we present a structure-function analysis of peptides derived from the human chemokines macrophage inflammatory protein-3alpha (MIP-3alpha/CCL20), interleukin 8 (IL-8), neutrophil activating protein-2 (NAP-2) and thrombocidin-1 (TC-1). These peptides encompass the C-terminal alpha-helices of these chemokines, which have been suggested to be important for the direct antimicrobial activities. Far UV CD spectroscopy showed that the peptides are unstructured in aqueous solution and that a membrane mimetic solvent is required to induce a helical secondary structure. A co-solvent mixture was used to determine solution structures of the peptides by two-dimensional (1)H-NMR spectroscopy. The highly cationic peptide, MIP-3alpha(51-70), had the most pronounced antimicrobial activity and displayed an amphipathic structure. A shorter version of this peptide, MIP-3alpha(59-70), remained antimicrobial but its structure and mechanism of action were unlike that of the former peptide. The NAP-2 and TC-1 proteins differ in their sequences only by the deletion of two C-terminal residues in TC-1, but intact TC-1 is a very potent antimicrobial while NAP-2 is inactive. The corresponding C-terminal peptides, NAP-2(50-70) and TC-1(50-68), had very limited and no bactericidal activity, respectively. This suggests that other regions of TC-1 contribute to its bactericidal activity. Altogether, this work provides a rational structural basis for the biological activities of these peptides and proteins and highlights the importance of experimental characterization of peptide fragments as distinct entities because their activities and structural properties may differ substantially from their parent proteins. PMID- 20004171 TI - Butyrylcholinesterase for protection from organophosphorus poisons: catalytic complexities and hysteretic behavior. AB - Butyrylcholinesterase is a promiscuous enzyme that displays complex kinetic behavior. It is toxicologically important because it detoxifies organophosphorus poisons (OP) by making a covalent bond with the OP. The OP and the butyrylcholinesterase are both inactivated in the process. Inactivation of butyrylcholinesterase has no adverse effects. However, inactivation of acetylcholinesterase in nerve synapses can be lethal. OP-inhibited butyrylcholinesterase and acetylcholinesterase can be reactivated with oximes provided the OP has not aged. Strategies for preventing the toxicity of OP include (a) treatment with an OP scavenger, (b) reaction of non-aged enzyme with oximes, (c) reactivation of aged enzyme, (d) slowing down aging with peripheral site ligands, and (e) design of mutants that rapidly hydrolyze OP. Option (a) has progressed through phase I clinical trials with human butyrylcholinesterase. Option (b) is in routine clinical use. The others are at the basic research level. Butyrylcholinesterase displays complex kinetic behavior including activation by positively charged esters, ability to hydrolyze amides, and a lag time (hysteresis) preceding hydrolysis of benzoylcholine and N-methylindoxyl acetate. Mass spectrometry has identified new OP binding motifs on tyrosine and lysine in proteins that have no active site serine. It is proposed, but not yet proven, that low dose exposure involves OP modification of proteins that have no active site serine. PMID- 20004173 TI - Differential effects of conjugated linoleic acid isomers on the biophysical and biochemical properties of model membranes. AB - Conjugated linoleic acids (CLA) are known to exert several isomer-specific biological effects, but their mechanisms of action are unclear. In order to determine whether the physicochemical effects of CLA on membranes play a role in their isomer-specific effects, we synthesized phosphatidylcholines (PCs) with 16:0 at sn-1 position and one of four CLA isomers (trans 10 cis 12 (A), trans 9 trans 11 (B), cis 9 trans 11 (C), and cis 9 cis 11 (D)) at sn-2, and determined their biophysical properties in monolayers and bilayers. The surface areas of the PCs with the two natural CLA (A and C) were similar at all pressures, but they differed significantly in the presence of cholesterol, with PC-A condensing more than PC-C. Liposomes of PC-A similarly showed increased binding of cholesterol compared to PC-C liposomes. PC-A liposomes were less permeable to carboxyfluorescein compared to PC-C liposomes. The PC with two trans double bonds (B) showed the highest affinity to cholesterol and lowest permeability. The two natural CLA-PCs (A and C) stimulated lecithin-cholesterol acyltransferase activity by 2-fold, whereas the unnatural CLA-PCs (B and D) were inhibitory. These results suggest that the differences in the biophysical properties of CLA isomers A and C may partly contribute to the known differences in their biological effects. PMID- 20004174 TI - Design of peptide-targeted liposomes containing nucleic acids. AB - Anticancer systemic gene silencing therapy has been so far limited by the inexistence of adequate carrier systems that ultimately provide an efficient intracellular delivery into target tumor cells. In this respect, one promising strategy involves the covalent attachment of internalizing-targeting ligands at the extremity of PEG chains grafted onto liposomes. Therefore, the present work aims at designing targeted liposomes containing nucleic acids, with small size, high encapsulation efficiency and able to be actively internalized by SCLC cells, using a hexapeptide (antagonist G) as a targeting ligand. For this purpose, the effect of the liposomal preparation method, loading material (ODN versus siRNA) and peptide-coupling procedure (direct coupling versus post-insertion) on each of the above-mentioned parameters was assessed. Post-insertion of DSPE-PEG antagonist G conjugates into preformed liposomes herein named as stabilized lipid particles, resulted in targeted vesicles with a mean size of about 130 nm, encapsulation efficiency close to 100%, and a loading capacity of approximately 5 nmol siRNA/mumol of total lipid. In addition, the developed targeted vesicles showed increased internalization in SCLC cells, as well as in other tumor cells and HMEC-1 microvascular endothelial cells. The improved cellular association, however, did not correlate with enhanced downregulation of the target protein (Bcl-2) in SCLC cells. These results indicate that additional improvements need to be performed in the future, namely by ameliorating the access of the nucleic acids to the cytoplasm of the tumor cells following receptor-mediated endocytosis. PMID- 20004175 TI - Interaction with membrane mimics of transmembrane fragments 16 and 17 from the human multidrug resistance ABC transporter 1 (hMRP1/ABCC1) and two of their tryptophan variants. AB - The human multidrug resistance-associated protein 1 (hMRP1/ABCC1) belongs to the ATP-binding cassette transporter superfamily. Together with P-glycoprotein (ABCB1) and the breast cancer resistance protein (BCRP/ABCG2), hMRP1 confers resistance to a large number of structurally diverse drugs. The current topological model of hMRP1 includes two cytosolic nucleotide-binding domains and 17 putative transmembrane (TM) helices forming three membrane-spanning domains. Mutagenesis and labeling studies have shown TM16 and TM17 to be important for function. We characterized the insertion of the TM16 fragment into dodecylphosphocholine (DPC) or n-dodecyl-beta-d-maltoside (DM) micelles as membrane mimics and extended our previous work on TM17 (Vincent et al., 2007, Biochim. Biophys. Acta 1768, 538). We synthesized TM16 and TM17, with the Trp residues, W1198 in TM16 and W1246 in TM17, acting as an intrinsic fluorescent probe, and TM16 and TM17 Trp variants, to probe different positions in the peptide sequence. We assessed the interaction of peptides with membrane mimics by evaluating the increase in fluorescence intensity resulting from such interactions. In all micelle-bound peptides, the tryptophan residue appeared to be located, on average, in the head group micelle region, as shown by its fluorescence spectrum. Each tryptophan residue was partially accessible to both acrylamide and the brominated acyl chains of two DM analogs, as shown by fluorescence quenching. Tryptophan fluorescence lifetimes were found to depend on the position of the tryptophan residue in the various peptides, probably reflecting differences in local structures. Far UV CD spectra showed that TM16 contained significant beta-strand structures. Together with the high Trp correlation times, the presence of these structures suggests that TM16 self association may occur at the interface. In conclusion, this experimental study suggests an interfacial location for both TM16 and TM17 in membrane mimics. In terms of overall hMRP1 structure, the experimentally demonstrated amphipathic properties of these TM are consistent with a role in the lining of an at least partly hydrophilic transport pore, as suggested by the currently accepted structural model, the final structure being modified by interaction with other TM helices. PMID- 20004176 TI - Glycyl betaine is effective in slowing down the irreversible denaturation of a detergent-solubilized membrane protein, sarcoplasmic reticulum Ca2+-ATPase (SERCA1a). AB - Many membrane proteins become labile when they are solubilized by detergent. Here we show that the presence of high concentrations of glycyl betaine stabilizes one of these proteins, the sarcoplasmic reticulum Ca(2+)-ATPase (SERCA1a), solubilized with nonionic detergents like n-dodecyl beta-d-maltopyranoside (DDM) or octaethylene glycol monododecyl ether (C(12)E(8)) which are commonly used for its purification or crystallization. Betaine at high concentrations might become useful as a stabilizing agent for detergent-solubilized membrane proteins, for instance during purification procedures or during the long periods of time required for crystallogenesis. PMID- 20004177 TI - Detection and characterization of side population in Ewing's sarcoma SK-ES-1 cells in vitro. AB - Dye exclusion is a valuable technique to isolate cancer stem cells (CSCs) based on an ability of stem cell to efflux fluorescent DNA-binding dye, especially for tumors without unique surface markers. It has been proven that side population (SP) cells that exclude Hoechst 33342 dye are enriched with stem-like cells in several cancer cell lines. In this study, we isolated and characterized SP cells from human Ewing's sarcoma cell line SK-ES-1 in vitro. SP cells were detected in SK-ES-1 and comprised 1.2% of total cell population. Only SP cells had the capacity to regenerate both SP and non-SP cells. The proliferation rates were similar between SP and non-SP cells. However, the clonogenicity and invasiveness of SP cells were significantly higher than that of non-SP cells. Further characterization of this SP phenotype presented other properties. SP cells exhibited increased multi-drug resistance and the ATP binding cassette protein (ABC) transporters were up-regulated in SP population. These findings suggest that SP cells derived from Ewing's sarcoma play the critical role in tumor metastasis and recurrence and might be an ideal target for clinical therapy. PMID- 20004178 TI - Cystatin M expression is reduced in gastric carcinoma and is associated with promoter hypermethylation. AB - Cystatin M is a secreted inhibitor of lysosomal cysteine proteases and increasing evidences indicate that it is a novel target for epigenetic silencing during mammary tumorigenesis. Aberrant promoter methylation is a well-known mechanism that participates in cystatin M silencing in breast cancer. However, the role of cystatin M in the gastric cancer remains to be elucidated. Immunohistochemistry was used to investigate the expression of cystatin M in 60 gastric carcinomas. Hypermethylation of cystatin M promoter was evaluated by the methylation-specific PCR (MSP) method in gastric carcinomas (tumor and paired adjacent non-tumor tissues). Reverse-transcriptase PCR and BSP were also performed on gastric cancer cell lines before and after treatment with 5-aza-2'-deoxycytidine (5-Aza-dC). Lost expression of cystatin M was observed in 42 of 60 (70%) gastric carcinomas. 55% (33 of 60) of primary tumors analyzed displayed cystatin M promoter hypermethylation, indicating that this aberrant characteristic is common in gastric malignancies. Moreover, a statistically significant inverse association was found between cystatin M methylation status and expression of the cystatin M protein in tumor tissues (p=0.027). We also found that patients with cystatin M promoter methylation had a significantly shorter survival time than those without this methylation (p=0.020). These results suggest that cystatin M promoter hypermethylation is one of the molecular mechanisms that accounts for reduced cystatin M gene expression in gastric carcinomas. PMID- 20004179 TI - Monocytes deliver bioactive nerve growth factor through a brain capillary endothelial cell-monolayer in vitro and counteract degeneration of cholinergic neurons. AB - Alzheimer's disease is an age-dependent brain disorder, characterized by progressive memory deficits and cognitive decline and loss of cholinergic neurons. Nerve growth factor (NGF) is the most potent protein to protect cholinergic neurons against degeneration. However, problems of delivery to the brain limit the therapeutical use of NGF. The aim of the present study was to test, if primary rat monocytes can be loaded with recombinant NGF and pass an in vitro monolayer of brain capillary endothelial cells (BCEC), release NGF, and support the cholinergic neurons in an organotypic brain slice model. Monocytes were isolated from rat blood by negative magnetic selection, loaded with recombinant NGF using Bioporter. The monocytes adhered and migrated through an in vitro rat BCEC-monolayer. NGF released at the basolateral side counteracted degeneration of cholinergic basal nucleus of Meynert neurons. In conclusion, our present study shows a proof-of-principle, that primary monocytes secreting NGF might be useful tools to deliver NGF into the brain, however, further in vivo studies are necessary. PMID- 20004181 TI - Attention inhibition of early cortical activation to fearful faces. AB - Several lines of evidence demonstrate that processing facial expression can occur in the first 130 ms following a face presentation, but it remains unclear how this is modulated by attention. We presented neutral, fearful and happy faces to subjects who attended either to repeated identity or to repeated emotions. Brain activity was recorded using magnetoencephalography (MEG) and analyzed with event related beamforming, providing both temporal and spatial information of processing in the brain. The first MEG component, at 90 ms (M90), was sensitive to facial expression, but only when attention was not directed to expression; non attended fearful faces increased activation in occipital and right middle frontal gyri. Around 150 ms, activity in several brain regions, regardless of the direction of attention, was larger to emotional compared to neutral faces; attention directed to facial expressions increased activity in the right fusiform gyrus and the anterior insula bilaterally. M220 was not modulated by individual facial expressions; however, attention directed to facial expressions enhanced activity in the right inferior parietal lobe and precuneus, while attention directed to identity enhanced posterior cingulate activity.These data demonstrate that facial expression processing involves frontal brain areas as early as 90 ms. Attention directed to emotional expressions obscured this early automatic processing but increased the M170 activity. The M220 sources varied with the direction of attention. Thus, the pattern of neural activation to faces varied with attention to emotions or to identity, demonstrating separate and only partially overlapping networks for these two facets of information contained in faces. PMID- 20004182 TI - The control of benzene exposure in North America, Europe and some Asian nations has been highly successful but industrial exposure in other countries continues to be of concern. Preface. PMID- 20004180 TI - Alterations of resting state functional connectivity in the default network in adolescents with autism spectrum disorders. AB - Autism spectrum disorders (ASD) are associated with disturbances of neural connectivity. Functional connectivity between neural structures is typically examined within the context of a cognitive task, but also exists in the absence of a task (i.e., "rest"). Connectivity during rest is particularly active in a set of structures called the default network, which includes the posterior cingulate cortex (PCC), retrosplenial cortex, lateral parietal cortex/angular gyrus, medial prefrontal cortex, superior frontal gyrus, temporal lobe, and parahippocampal gyrus. We previously reported that adults with ASD relative to controls show areas of stronger and weaker connectivity within the default network. The objective of the present study was to examine the default network in adolescents with ASD. Sixteen adolescents with ASD and 15 controls participated in a functional MRI study. Functional connectivity was examined between a PCC seed and other areas of the default network. Both groups showed connectivity in the default network. Relative to controls, adolescents with ASD showed widespread weaker connectivity in nine of the eleven areas of the default network. Moreover, an analysis of symptom severity indicated that poorer social skills and increases in restricted and repetitive behaviors and interests correlated with weaker connectivity, whereas poorer verbal and non-verbal communication correlated with stronger connectivity in multiple areas of the default network. These findings indicate that adolescents with ASD show weaker connectivity in the default network than previously reported in adults with ASD. The findings also show that weaker connectivity within the default network is associated with specific impairments in ASD. PMID- 20004183 TI - Purification of a peptide from seahorse, that inhibits TPA-induced MMP, iNOS and COX-2 expression through MAPK and NF-kappaB activation, and induces human osteoblastic and chondrocytic differentiation. AB - Ongoing efforts to search for naturally occurring, bioactive substances for the amelioration of arthritis have led to the discovery of natural products with substantial bioactive properties. The seahorse (Hippocampus kuda Bleeler), a telelost fish, is one source of known beneficial products, yet has not been utilized for arthritis research. In the present work, we have purified and characterized a bioactive peptide from seahorse hydrolysis. Among the hydrolysates tested, pronase E-derived hydrolysate exhibited the highest alkaline phosphatase (ALP) activity, a phenotype marker of osteoblast and chondrocyte differentiation. After its separation from the hydrolysate by several purification steps, the peptide responsible for the ALP activity was isolated and its sequence was identified as LEDPFDKDDWDNWK (1821Da). We have shown that the isolated peptide induces differentiation of osteoblastic MG-63 and chondrocytic SW-1353 cells by measuring ALP activity, mineralization and collagen synthesis. Our results indicate that the peptide acts during early to late stages of differentiation in MG-63 and SW-1353 cells. We also assessed the concentration dependence of the peptide's inhibition of MMP (-1, -3 and -13), iNOS and COX-2 expression after treatment with 12-O-tetradecanoylphorbol-13-acetate (TPA), a common form of phorbol ester. The peptide also inhibited NO production in MG-63 and SW-1353 cells. To elucidate the mechanisms by which the peptide acted, we examined its effects on TPA-induced MAPKs/NF-kappaB activation and determined that the peptide treatment significantly reduced p38 kinase/NF-kappaB in MG-63 cells and MAPKs/NF-kappaB in SW-1353 cells. PMID- 20004184 TI - Benzene 2009--Health effects and mechanisms of bone marrow toxicity: implications for t-AML and the mode of action framework. AB - This overview of the Symposium and its organization includes a historical survey of the scientific literature in which the relationship of benzene exposure to the development of aplastic anemia and other bone marrow diseases, including acute myeloid (myelogenous) leukemia, is described. Previous conferences on the health effects of benzene are summarized. The important role of the revised World Health Organization classification of tumors of the hematopoietic and lymphoid tissues in clarifying the specific diseases related to benzene exposure is emphasized. PMID- 20004185 TI - 25-OH-vitamin D assay variation and subject management in clinical practice. AB - OBJECTIVE: To compare two methods for assessing vitamin D status in clinical settings. DESIGN AND METHODS: 25-OH-vitamin D status was measured in 120 patients by HPLC and a commercial immunoassay. RESULTS: Only 53% of the subjects considered as vitamin D sufficient by immunoassay fell within this category according to HPLC although subjects with concentrations above 75 nmol/L, regardless of the method used, presented normal PTH concentrations. CONCLUSIONS: Both methods are not exchangeable to classify subjects based on unique cut-offs but they are comparable when interpreted in relation with PTH concentrations. PMID- 20004186 TI - Initiation of neuronal differentiation requires PI3-kinase/TOR signalling in the vertebrate neural tube. AB - Regulated neuron production within the vertebrate nervous system relies on input from multiple signalling pathways. Work in the Drosophila retina has demonstrated that PI3-kinase and downstream TOR signalling regulate the timing of photoreceptor differentiation; however, the function of such signals during vertebrate neurogenesis is not well understood. Here we show that mutant mice lacking PKB activity downstream of PDK1, the master kinase of the PI3-kinase pathway, exhibit deficient neuron production. We further demonstrate expression of PI3-kinase signalling components and active PKB and TOR signalling in the chick spinal cord, an early site of neurogenesis. Neuron production was also attenuated in the chick neural tube following exposure to small molecule inhibitors of PI3-kinase (LY294002) or TOR (Rapamycin) activity. Furthermore, Rapamycin repressed expression of early neuronal differentiation genes, such as Ngn2, but did not inhibit expression of Sox1B genes characteristic of proliferating neural progenitors. In addition, some cells expressing an early neuronal marker were mis-localised at the ventricular surface in the presence of Rapamycin and remained aberrantly within the cell cycle. These findings suggest that TOR signalling is necessary to initiate neuronal differentiation and that it may facilitate coordination of cell cycle and differentiation programmes. In contrast, stimulating PI3-kinase signalling did not increase neuron production, suggesting that such activity is simply permissive for vertebrate neurogenesis. PMID- 20004187 TI - Intestinal tube formation in Caenorhabditis elegans requires vang-1 and egl-15 signaling. AB - Understanding how epithelial organs form during morphogenesis is a major problem in developmental biology. In the present paper, we provide a detailed analysis of vang-1, the only homolog of the planar cell polarity protein Strabismus/Van Gogh in Caenorhabditis elegans. We demonstrate that during organogenesis of the intestine, (i) VANG-1 specifically interacts with PDZ 2 domain of DLG-1 (Discs large) and becomes phosphorylated by the kinase domain of the FGF-like receptor tyrosine kinase EGL-15; (ii) VANG-1 is predominantly restrained to the cell cortex but relocates to the apical junction; and (iii) in vang-1 embryos epithelial cells of the intestine are not correctly arranged along the anterior posterior axis. To investigate what determines the disposition of the VANG-1 protein, either truncated protein forms were expressed in the intestine or RNAi was used to remove the functions of gene products previously shown to be involved in apical junction formation. Removal of the VANG-1 PDZ binding motif "-ESAV" and depletion of dlg-1 or let-413 gene functions interferes with the localization of VANG-1. In addition, egl-15 embryos show a premature relocation of VANG-1 to the apical junction, causing defects that resemble those observed in mutant vang-1 embryos and after intestine-specific overexpression of full-length vang-1. Finally, the localization of VANG-1 depends on DSH-2, a homolog of the planar cell polarity protein Dishevelled and depletion phenocopies vang-1 and egl-15 phenotypes in the embryonic intestine. PMID- 20004188 TI - Purkinje cells originate from cerebellar ventricular zone progenitors positive for Neph3 and E-cadherin. AB - GABAergic Purkinje cells (PCs) provide the primary output from the cerebellar cortex, which controls movement and posture. Although the mechanisms of PC differentiation have been well studied, the precise origin and initial specification mechanism of PCs remain to be clarified. Here, we identified a cerebellar and spinal cord GABAergic progenitor-selective cell surface marker, Neph3, which is a direct downstream target gene of Ptf1a, an essential regulator of GABAergic neuron development. Using FACS, Neph3(+) GABAergic progenitors were sorted from the embryonic cerebellum, and the cell fate of this population was mapped by culturing in vitro. We found that most of the Neph3(+) populations sorted from the mouse E12.5 cerebellum were fated to differentiate into PCs while the remaining small fraction of Neph3(+) cells were progenitors for Pax2(+) interneurons, which are likely to be deep cerebellar nuclei GABAergic neurons. These results were confirmed by short-term in vivo lineage-tracing experiments using transgenic mice expressing Neph3 promoter-driven GFP. In addition, we identified E-cadherin as a marker selectively expressed by a dorsally localized subset of cerebellar Neph3(+) cells. Sorting experiments revealed that the Neph3(+) E-cadherin(high) population in the embryonic cerebellum defined PC progenitors while progenitors for Pax2(+) interneurons were enriched in the Neph3(+) E-cadherin(low) population. Taken together, our results identify two spatially demarcated subregions that generate distinct cerebellar GABAergic subtypes and reveal the origin of PCs in the ventricular zone of the cerebellar primordium. PMID- 20004189 TI - 17beta-Estradiol decreases vulnerability to ventricular arrhythmias by preserving connexin43 protein in infarcted rats. AB - Epidemiological studies showed that a lower mortality rate of sudden cardiac death among women than among men may depend on the action of female sex hormones. This study assessed whether 17beta-estradiol exerts anti-arrhythmic effects through enhanced Connexin43 (Cx43) expression after infarction. Two weeks after ovariectomy, female Wistar rats were randomly assigned to coronary artery ligation or sham-operation. Twenty-four hours after coronary ligation, ovariectomized rats were randomized into vehicle, subcutaneous estradiol treatment, tamoxifen, or subcutaneous estradiol treatment+tamoxifen and followed for 4weeks. To verify the role of estradiol-related nitric oxide in modulating the expression of Cx43, N-nitro-L-arginine methyl ester was also assessed in an in vitro study. Myocardial Cx43 expression revealed a significant decrease in vehicle-treated infarcted rats compared with sham-operated rats at 24h and 4weeks after infarction. Attenuated Cx43 expression was blunted after administering estradiol, assessed by immunofluorescent analysis, Western blotting, and real time quantitative RT-PCR of Cx43. The vulnerability for ventricular arrhythmia during programmed stimulation in estradiol-treated infarcted rats was significantly lower than in vehicle-treated infarcted rats. The beneficial effect of estradiol on Cx43 was abolished by tamoxifen. In addition, the invitro study demonstrated that the amount of Cx43 showed significant reduction after adding N nitro-L-arginine methyl ester. Chronic administration of estradiol after infarction is associated with attenuated reduction of gap junction proteins probably through a nitric oxide-dependent pathway via the estrogen receptor and thus plays a critical role in the beneficial effect on arrhythmic vulnerability response to programmed electrical stimulation. PMID- 20004190 TI - A comparative study of alpha-adrenergic receptor mediated Ca(2+) signals and contraction in intact human and mouse vascular smooth muscle. AB - In many vascular smooth muscle cells, physiological and pharmacological agonists initiate oscillatory fluctuations in intracellular Ca(2+) to initiate and maintain vasoconstriction. These oscillations are supported by the underlying cellular ultrastructure, particularly the close apposition between the plasma membrane (PM) and superficial sarcoplasmic reticulum (SR), the so-called PM-SR junctions, which are important for SR Ca(2+) refilling. We hypothesize that the disappearance of PM-SR junctions during aging and/or disease is directly related to the disappearance of agonist-induced Ca(2+) oscillations. We compared phenylephrine-mediated Ca(2+) signals and contraction in human and murine smooth muscle cells in small mesenteric arteries and also employed electron microscopy to examine the cytoplasmic distribution of the SR. Phenylephrine elicited tonic contractions in both types of vessels, asynchronous Ca(2+) oscillations in the mouse mesenteric smooth muscle cells, but only single transient Ca(2+) signals in the human mesenteric smooth muscle cells. While nifedipine inhibited 90% of the phenylephrine-induced tonic contraction in mouse mesenteric arteries, it only slightly attenuated tonic contraction in human mesenteric arteries, although the nifedipine-resistant component was abolished by the Rho-kinase blocker 1-(5 Isoquinolinylsulfonyl)homopiperazine dihydrochloride (HA-1077). Furthermore, superficial SR was found to be abundant in the mouse vessels and many PM-SR junctions were observed, but the smooth muscle of human mesenteric arteries had far less peripheral SR and was almost devoid of PM-SR junctions. As PM-SR junctions are essential for the maintenance of Ca(2+) oscillations, the change in Ca(2+) signalling pattern in the relatively old human patients was due to impaired SR refilling. PMID- 20004191 TI - YB1/p32, a nuclear Y-box binding protein 1, is a novel regulator of myoblast differentiation that interacts with Msx1 homeoprotein. AB - Precisely controlled cellular differentiation is essential for the proper development of vertebrate embryo and deregulated differentiation is a major cause of many human congenital diseases as well as cancer. Msx1 is a member of the homeoprotein family implicated in these processes, which inhibits the differentiation of skeletal muscle and other cell types, presumably by regulating transcription of target genes through interaction with other cellular factors. We presently show that YB1/p32, a nuclear Y-box binding protein 1, interacts with Msx1 homeoprotein and functions as a regulator of C2C12 myoblast differentiation. We demonstrate that YB1/p32 functionally interacts with Msx1 through its N terminal region and colocalizes with Msx1 at the nuclear periphery. Moreover, we find that YB1/p32 is competent for inhibition of C2C12 myoblast differentiation, which is correlated with its activity as a negative regulator of MyoD gene expression and binding to the MyoD core enhancer region (CER). Furthermore, YB1/p32 cooperates with Msx1 in transcriptional repression and knocking down the expression of endogenous YB1 attenuates the effects of Msx1. Taken together, our study has uncovered a new function of YB1/p32, a regulator of skeletal muscle differentiation. PMID- 20004192 TI - Glial cell line-derived neurotrophic factor alters the growth characteristics and genomic imprinting of mouse multipotent adult germline stem cells. AB - This study evaluated the essentiality of glial cell line-derived neurotrophic factor (GDNF) for in vitro culture of established mouse multipotent adult germline stem (maGS) cell lines by culturing them in the presence of GDNF, leukemia inhibitory factor (LIF) or both. We show that, in the absence of LIF, GDNF slows the proliferation of maGS cells and result in smaller sized colonies without any change in distribution of cells to different cell-cycle stages, expression of pluripotency genes and in vitro differentiation potential. Furthermore, in the absence of LIF, GDNF increased the expression of male germ line genes and repopulated the empty seminiferous tubule of W/W(v) mutant mouse without the formation of teratoma. GDNF also altered the genomic imprinting of Igf2, Peg1, and H19 genes but had no effect on DNA methylation of Oct4, Nanog and Stra8 genes. However, these effects of GDNF were masked in the presence of LIF. GDNF also did not interfere with the multipotency of maGS cells if they are cultured in the presence of LIF. In conclusion, our results suggest that, in the absence of LIF, GDNF alters the growth characteristics of maGS cells and partially impart them some of the germline stem (GS) cell-like characteristics. PMID- 20004193 TI - Distinct central representations for sensory fibers innervating either the conjunctiva or cornea of the rat. AB - The laminar sheet of epithelium (e.g., skin and mucous membrane) enclosing our bodies is represented in the dorsal horns of the medulla and spinal cord. The eyeball however indents this laminar sheet and is shrouded by different layers: the cornea/sclera, the conjunctiva, and hairy skin. This involution of the orb confounds defining the central representation of the cornea and its surrounding mucosa and skin. We used herein the transganglionic transport of a cocktail of HRP conjugated to cholera toxin and wheat germ agglutinin to determine the central representation of these epithelia in the dorsal horns of the rat. The HRP cocktail was injected either into the stroma of the cornea, the mucosa of the conjunctiva, or the supraorbital and infraorbital nerves. Injections of the cornea produced dense label in the interstitial islands in the ventral medullary dorsal horn (MDH), probably lamina I, and in neuropil in the ventromedial tip of the MDH, probably lamina II. There sometimes was variable, diffuse label in the C1 dorsal horn after corneal injections but more rostral parts of the trigeminal sensory complex were never labeled. Injections of the conjunctiva densely labeled laminae I-III in the C1 dorsal horn, while laminae IV-V were diffusely labeled. Sparser reaction product also was seen in lamina I in positions similar to the cornea projection. Label was seen ventrally in subnuclei interpolaris and oralis, as well as the principal trigeminal nucleus. Projections of the infraorbital nerve included all laminae in the trigeminocervical complex as well as large portions of the rostral subnuclei in the spinal trigeminal nucleus. The projections of the supraorbital nerve were similar, but were restricted to ventral parts of the trigeminal sensory complex. In other cases the cornea was injected either after cutting the supraorbital and infraorbital nerves or the conjunctiva was injected after enucleating the eyeball. Any reaction product from corneal injections was reduced dramatically in the C1 dorsal horn after transection of the infraorbital and supraorbital nerves. Injecting the conjunctiva after enucleating the eyeball densely labeled the C1 projection to the dorsal horn, a small patch in lamina I in the MDH, as well as the rostral trigeminal complex. We propose that the cornea has but a single representation in the trigeminocervical complex in its ventral part near the caudal end of the medulla. We also propose the palpebral conjunctiva mucosa is represented in the C1 dorsal horn, and speculate that the bulbar conjunctiva overlaps with that of the cornea in lamina I. We discuss these projections in relation to the circuitry for the supraorbital-evoked and corneal-evoked blink reflexes. The relationship of the cornea and conjunctiva is intimate, and investigators must be very careful when attempting to stimulate them in isolation. PMID- 20004194 TI - Echinococcus granulosus genotypes in livestock of Iran indicating high frequency of G1 genotype in camels. AB - In this study, 112 Echinococcus granulosus isolates from different livestock of Iran were genotyped by PCR amplification of ribosomal DNA-internal transcribed spacer 1 (rDNA-ITS1) region followed by restriction fragment length polymorphism (RFLP) with the enzyme RsaI. The possibility of intra-genotype variation was also investigated using randomly amplified polymorphic DNA (RAPD) analysis. Isolates from sheep, goats, cattle and the majority of camels (12 of 18; 66.7%) were identified as the G1 genotype and a few camel isolates (6 of 18; 33.3%) belonged to the G6 genotype. Overall G1 and G6 genotypes were identified in 94.6% (106 of 112) and 5.3% (6 of 112) of all isolates, respectively. RAPD analysis based on 15 separate primers showed 7-14 bands of 200-3000bp for strain G1. Considering each individual primer, no differences observed among isolates from different hosts and between livers and lungs. This study confirmed the existence of G1 and G6 genotypes in Iran. Moreover, G1 is much more prevalent even in camels, indicating the importance of sheep-dog cycle in public health. Studying intra-genotypic variation of E. granulosus warrants more research using other primers and methods. PMID- 20004195 TI - The reproductive output of sea lice Caligus rogercresseyi under controlled conditions. AB - Gravid females of Caligus rogercresseyi were collected from Atlantic salmon (Salmo salar) from a farm located at Chiloe Island (42 degrees 40'S73 degrees 15'W), Chile, to obtain information about the reproductive output of this parasite in vitro. The egg strings removed from the females were incubated under controlled conditions to obtain virgin adult females. One female which had mated only once produced eleven generations of eggs strings in a period of 74 days. The first egg strings of the females obtained in vitro were produced at 389 degree days ( degrees D) after egg incubation, while the next generations of eggs strings were produced with a periodicity between 4 and 6 days dependent on the water temperature. The average length of the egg string was 3.1mm and the mean number of eggs per string was 31. The values recorded in captivity for the egg string length and the number of eggs per string, were lower than the values recorded in gravid females from the field. One female survived for 79 days and males, maintained separately from the females, survived for 60 days. PMID- 20004196 TI - The genus Sappinia: history, phylogeny and medical relevance. AB - The genus Sappinia with the single species Sappinia pedata was established for an amoeba with two nuclei and pedicellate "cysts" by Dangeard in 1896. In 1912, Alexeieff transferred an also double nucleated, but apparently sexually reproducing amoeba to this genus as Sappinia diploidea, that had been described as Amoeba diploidea by Hartmann and Nagler in 1908. As the original isolates were lost, Michel and colleagues established a neotype for S. diploidea in 2006 and Brown and colleagues established a neotype for S. pedata in 2007. Molecular analyses have corroborated the differentiation between S. pedata and S. diploidea, however, the genus splits into more than two well separated clusters. Altogether, the genus Sappinia is now classified as a member of the Thecamoebidae and, moreover, as potentially pathogenic. In 2001, Gelman and colleagues reported a case of severe encephalitis in a non-immunocompromised young man caused by Sappinia. PMID- 20004197 TI - A special issue on parasitic crustaceans. PMID- 20004198 TI - Obox4 regulates the expression of histone family genes and promotes differentiation of mouse embryonic stem cells. AB - Obox genes are preferentially expressed in the ovary, testis and oocyte, and play important roles in many developmental processes. In this study, we report that Obox4 and Obox6 are expressed in mouse embryonic stem cells (mESCs) and that Obox4 regulates histone family gene expression in mESCs. Obox4 protein expressing mESCs formed colonies with a flattened and irregular morphology, and exhibited decreased expression levels of self-renewal related proteins, such as Oct4 and Sox2, as well as reduced alkaline phosphatase activity. The results of microarray analysis and siRNA mediated knockdown experiments suggest that Obox4 is an upstream regulator of the histone gene family. PMID- 20004199 TI - Structural requirements of anthocyanins in relation to inhibition of endothelial injury induced by oxidized low-density lipoprotein and correlation with radical scavenging activity. AB - Anthocyanins may play an important role in atherosclerosis prevention. However, the structure-function relationships are not well understood. The objective of this study was to compare the inhibitory effect of 21 anthocyanins against oxidized low-density lipoprotein-induced endothelial injury to understand the relationship between anthocyanin chemical structure and the endothelial protective properties, measured as cell viability, MDA production and NO release. Additionally, the intracellular anti-radical activity of the selected anthocyanins was investigated to identify the correlation with endothelial protection. Our results provide evidence that the number of -OH in total or in B ring, 3',4'-ortho-dihydroxyl and 3-hydroxyl are the main structural requirements of anthocyanins in suppressing oxidative stress-induced endothelial injury and such inhibitory effect was significantly correlated with the intracellular radical scavenging activity. PMID- 20004200 TI - Activation of oxidative stress response by hydroxyl substituted chalcones and cyclic chalcone analogues in mitochondria. AB - We investigated the effect of hydroxyl substituted chalcone (1a) and some chalcone analogues (1b-d) on isolated rat liver mitochondria to gain new insights into the cytotoxic mechanism of these compounds. We observed an inhibitory effect on phosphorylation and the partial uncoupling of compounds 1a and 1d. Increased radical generation and possible covalent interaction of the compounds with cellular thiols resulted in glutathione (GSH) depletion and modulation of the investigated mitochondrial activities. Disruption of interconnected mechanisms as electron transport chain and energetic metabolism, ROS production and insufficiency of antioxidant defensive system could lead to induction of cell death. PMID- 20004201 TI - Phosphatidylinositol 3-kinase gamma inhibition ameliorates inflammation and tumor growth in a model of colitis-associated cancer. AB - BACKGROUND & AIMS: A large body of evidence supports a correlation between inflammation and cancer, although the molecular mechanisms that govern this process are incompletely understood. Phosphatidylinositol 3-kinase (PI3K) is an enzyme that regulates the immune response and contributes to cell transformation in several tumor types. Here, we address the role of the PI3Kgamma isoform in inflammatory bowel disease and in the development of colitis-associated cancer. METHODS: PI3Kgamma(-/-) and control mice were repeatedly treated with dextran sulfate sodium to induce chronic colitis and colitis-associated cancer. Colorectal tumor burden and colon inflammation were evaluated in these mice. Leukocyte populations in colon were characterized by flow cytometry analysis. RESULTS: PI3Kgamma-deficient mice had a lower incidence of colitis-associated tumors, as well as reduced tumor multiplicity and smaller tumor size compared with controls. Reduced tumor development paralleled less colon inflammation in PI3Kgamma-deficient mice. Analysis of leukocyte populations in the colon of PI3Kgamma-deficient mice showed defective activation and infiltration of myeloid cells and defective recruitment of T cells to the colon compared with controls. CONCLUSIONS: PI3Kgamma regulates the innate immune response in a murine model of ulcerative colitis, thereby controlling colon inflammation and tumor formation. PMID- 20004202 TI - Tumor necrosis factor and interferon-gamma down-regulate Klotho in mice with colitis. AB - BACKGROUND & AIMS: Klotho (KL) is an anti-inflammatory protein that protects the endothelium from nitric oxide (NO)-induced dysfunction, reduces the expression of endothelial adhesion molecules, and potentially regulates T-cell functions. KL deficiency leads to premature senescence and impaired Ca2+/Pi homeostasis, which can lead to inflammatory bowel disease (IBD)-associated osteopenia/osteoporosis. We investigated the changes in renal expression of Kl as a consequence of colitis. METHODS: We studied 3 mouse models of IBD: colitis induced by trinitrobenzene sulfonic acid, colitis induced by microflora (in gnotobiotic interleukin-10(-/-)), and colitis induced by adoptive transfer of CD4(+)CD45RB(high) T cells. Effects of the tumor necrosis factor (TNF) and interferon (IFN)-gamma on Kl expression and the activity of its promoter were examined in renal epithelial cells (mpkDCT4 and mIMCD3). RESULTS: Renal expression of Kl messenger RNA (mRNA) and protein was reduced in all 3 models of IBD. Reduced level of KL correlated with the severity of colitis; the effect was reversed by neutralizing antibodies against TNF. In vitro, TNF inhibited Kl expression, an effect potentiated by IFN-gamma. The combination of TNF and IFN gamma increased expression of inducible nitric oxide synthase (iNOS) and increased NO production. The effect of IFN-gamma was reproduced by exposure to an NO donor and reversed by the iNOS inhibitor. In cells incubated with TNF and/or IFN-gamma, Kl mRNA stability was unaffected, whereas Kl promoter activity was reduced, indicating that these cytokines regulate Kl at the transcriptional level. CONCLUSIONS: The down-regulation of KL that occurs during inflammation might account for the extraintestinal complications such as abnormalities in bone homeostasis that occur in patients with IBD. PMID- 20004203 TI - Annual and periovulatory changes in tyrosine hydroxylase activity in the ovary of the catfish Heteropneustes fossilis. AB - In the present study, tyrosine hydroxylase (TH, the rate-limiting enzyme in catecholamine synthesis) activity was demonstrated in the ovary of the catfish to elucidate the possible physiological role of catecholamines in the gonad. The ovary is innervated by seven pairs of nerves, originating from the paired sympathetic chain lying dorsal to the posterior kidney. Ovarian TH activity showed a significant annual variation (P<0.001, one-way ANOVA), correlating with gonado-somatic index. Activity was low from December to February (resting phase), increased from March to July (recrudescent phase) and then decreased in post spawning phase (August-November). The annual pattern was similar to that of the brain. An intraperitoneal injection of 100 IU hCG/fish induced significant periovulatory changes in TH activity with the peak rise at 16 h, and the activity decreased after egg-stripping (P<0.001, one-way ANOVA). Brain TH activity showed similar periovulatory changes. The results suggest that catecholamine synthesis is increased during both ovarian recrudescence and spawning of the annual reproductive cycle, implying a functional role in ovarian growth, maturation and ovulation. PMID- 20004204 TI - A deficiency in the B cell response of C57BL/6 mice correlates with loss of macrophage-mediated killing of Leishmania amazonensis. AB - Infection of C3HeB/FeJ and C57BL/6 mice with Leishmania major stimulates a healing cell-mediated immune response, while Leishmania amazonensis infection leads to chronic disease. Here we show C3HeB/FeJ mice co-infected with both species of Leishmania heal, while co-infected C57BL/6 mice do not. Using an in vitro killing assay we determined B cells from infected C57BL/6 mice are ineffective in promoting parasite killing compared with B cells from infected C3HeB/FeJ mice. Furthermore, infected C57BL/6 mice produce less antigen-specific antibodies compared with infected C3HeB/FeJ mice. These findings suggest B cells play a required role in the cell-mediated immune response against L. amazonensis. PMID- 20004205 TI - Increase in backbone mobility of the VTS1p-SAM domain on binding to SRE-RNA. AB - The sterile alpha motif (SAM) domain of VTS1p, a posttranscriptional gene regulator, belongs to a family of SAM domains conserved from yeast to humans. Even though SAM domains were originally classified as protein-protein interaction domains, recently, it was shown that the yeast VTS1p-SAM and the SAM domain of its Drosophila homolog Smaug can specifically recognize RNA hairpins termed Smaug recognition element (SRE). Structural studies of the SRE-RNA complex of VTS1p-SAM revealed that the SAM domain primarily recognizes the shape of the RNA fold induced by the Watson-Crick base-pairing in the RNA pentaloop. Only the central G nucleotide is specifically recognized. The VTS1p-SAM domain recognizes SRE-RNAs with a CNGGN pentaloop where N is any nucleotide. The C1-G4 base pair in the wild type can be replaced by any pair of nucleotides that can form base pairs even though the binding affinity is greatest with a pyrimidine in position 1 and a purine in position 4. The interaction thus combines elements of sequence-specific and non-sequence-specific recognitions. The lack of structural rearrangements in either partner following binding is rather intriguing, suggesting that molecular dynamics may play an important role in imparting relaxed specificity with respect to the exact combination of nucleotides in the loop, except for the central nucleotide. In this work, we extend our previous studies of SRE-RNA interaction with VTS1p, by comparing the dynamics of the VTS1p-SAM domain both in its free form and when bound to SRE-RNA. The 15N relaxation studies of backbone dynamics suggest the presence of a dynamic interaction interface, with residues associated with specific G3 recognition becoming more rigid on RNA binding while other regions attain increased flexibility. The results parallel the observations from our studies of dynamics changes in SRE-RNA upon binding to VTS1p-SAM and shows that molecular dynamics could play a crucial role in modulating binding affinity and possibly contribute to the free energy of the interaction through an entropy driven mechanism. PMID- 20004206 TI - Light activation of rhodopsin: insights from molecular dynamics simulations guided by solid-state NMR distance restraints. AB - Structural restraints provided by solid-state NMR measurements of the metarhodopsin II intermediate are combined with molecular dynamics simulations to help visualize structural changes in the light activation of rhodopsin. Since the timescale for the formation of the metarhodopsin II intermediate (>1 ms) is beyond that readily accessible by molecular dynamics, we use NMR distance restraints derived from 13C dipolar recoupling measurements to guide the simulations. The simulations yield a working model for how photoisomerization of the 11-cis retinylidene chromophore bound within the interior of rhodopsin is coupled to transmembrane helix motion and receptor activation. The mechanism of activation that emerges is that multiple switches on the extracellular (or intradiscal) side of rhodopsin trigger structural changes that converge to disrupt the ionic lock between helices H3 and H6 on the intracellular side of the receptor. PMID- 20004208 TI - Characterization of the head-to-tail overlap complexes formed by human lamin A, B1 and B2 "half-minilamin" dimers. AB - Half-minilamins, representing amino- and carboxy-terminal fragments of human lamins A, B1 and B2 with a truncated central rod domain, were investigated for their ability to form distinct head-to-tail-type dimer complexes. This mode of interaction represents an essential step in the longitudinal assembly reaction exhibited by full-length lamin dimers. As determined by analytical ultracentrifugation, the amino-terminal fragments were soluble under low ionic strength conditions sedimenting with distinct profiles and s-values (1.6-1.8 S) indicating the formation of coiled-coil dimers. The smaller carboxy-terminal fragments were, except for lamin B2, largely insoluble under these conditions. However, after equimolar amounts of homotypic amino- and carboxy-terminal lamin fragments had been mixed in 4 M urea, upon subsequent renaturation the carboxy terminal fragments were completely rescued from precipitation and distinct soluble complexes with higher s-values (2.3-2.7 S) were obtained. From this behavior, we conclude that the amino- and carboxy-terminal coiled-coil dimers interact to form distinct oligomers (i.e. tetramers). Furthermore, a corresponding interaction occurred also between heterotypic pairs of A- and B type lamin fragments. Hence, A-type lamin dimers may interact with B-type lamin dimers head-to-tail to yield linear polymers. These findings indicate that a lamin dimer principally has the freedom for a "combinatorial" head-to-tail association with all types of lamins, a property that might be of significant importance for the assembly of the nuclear lamina. Furthermore, we suggest that the head-to-tail interaction of the rod end domains represents a principal step in the assembly of cytoplasmic intermediate filament proteins too. PMID- 20004207 TI - Combination of the loss of cmnm5U34 with the lack of s2U34 modifications of tRNALys, tRNAGlu, and tRNAGln altered mitochondrial biogenesis and respiration. AB - Yeast Saccharomyces cerevisiae MTO2, MTO1, and MSS1 genes encoded highly conserved tRNA modifying enzymes for the biosynthesis of carboxymethylaminomethyl (cmnm)(5)s(2)U(34) in mitochondrial tRNA(Lys), tRNA(Glu), and tRNA(Gln). In fact, Mto1p and Mss1p are involved in the biosynthesis of the cmnm(5) group (cmnm(5)U(34)), while Mto2p is responsible for the 2-thiouridylation (s(2)U(34)) of these tRNAs. Previous studies showed that partial modifications at U(34) in mitochondrial tRNA enabled mto1, mto2, and mss1 strains to respire. In this report, we investigated the functional interaction between MTO2, MTO1, and MSS1 genes by using the mto2, mto1, and mss1 single, double, and triple mutants. Strikingly, the deletion of MTO2 was synthetically lethal with a mutation of MSS1 or deletion of MTO1 on medium containing glycerol but not on medium containing glucose. Interestingly, there were no detectable levels of nine tRNAs including tRNA(Lys), tRNA(Glu), and tRNA(Gln) in mto2/mss1, mto2/mto1, and mto2/mto1/mss1 strains. Furthermore, mto2/mss1, mto2/mto1, and mto2/mto1/mss1 mutants exhibited extremely low levels of COX1 and CYTB mRNA and 15S and 21S rRNA as well as the complete loss of mitochondrial protein synthesis. The synthetic enhancement combinations likely resulted from the completely abolished modification at U(34) of tRNA(Lys), tRNA(Glu), and tRNA(Gln), caused by the combination of eliminating the 2-thiouridylation by the mto2 mutation with the absence of the cmnm(5)U(34) by the mto1 or mss1 mutation. The complete loss of modifications at U(34) of tRNAs altered mitochondrial RNA metabolisms, causing a degradation of mitochondrial tRNA, mRNA, and rRNAs. As a result, failures in mitochondrial RNA metabolisms were responsible for the complete loss of mitochondrial translation. Consequently, defects in mitochondrial protein synthesis caused the instability of their mitochondrial genomes, thus producing the respiratory-deficient phenotypes. Therefore, our findings demonstrated a critical role of modifications at U(34) of tRNA(Lys), tRNA(Glu), and tRNA(Gln) in maintenance of mitochondrial genome, mitochondrial RNA stability, translation, and respiratory function. PMID- 20004210 TI - Estimating trees from filtered data: identifiability of models for morphological phylogenetics. AB - As an alternative to parsimony analyses, stochastic models have been proposed (Lewis, 2001; Nylander et al., 2004) for morphological characters, so that maximum likelihood or Bayesian analyses may be used for phylogenetic inference. A key feature of these models is that they account for ascertainment bias, in that only varying, or parsimony-informative characters are observed. However, statistical consistency of such model-based inference requires that the model parameters be identifiable from the joint distribution they entail, and this issue has not been addressed. Here we prove that parameters for several such models, with finite state spaces of arbitrary size, are identifiable, provided the tree has at least eight leaves. If the tree topology is already known, then seven leaves suffice for identifiability of the numerical parameters. The method of proof involves first inferring a full distribution of both parsimony informative and non-informative pattern joint probabilities from the parsimony informative ones, using phylogenetic invariants. The failure of identifiability of the tree parameter for four-taxon trees is also investigated. PMID- 20004209 TI - Identification of novel contributions to high-affinity glycoprotein-receptor interactions using engineered ligands. AB - Engineered receptor fragments and glycoprotein ligands employed in different assay formats have been used to dissect the basis for the dramatic enhancement of binding of two model membrane receptors, dendritic cell-specific intercellular adhesion molecule 3-grabbing nonintegrin (DC-SIGN) and the macrophage galactose lectin, to glycoprotein ligands compared to simple sugars. These approaches make it possible to quantify the importance of two major factors that combine to enhance the affinity of single carbohydrate-recognition domains (CRDs) for glycoprotein ligands by 100-to 300-fold. First, the presence of extended binding sites within a single CRD can enhance interaction with branched glycans, resulting in increases of fivefold to 20-fold in affinity. Second, presentation of glycans on a glycoprotein surface increases affinity by 15-to 20-fold, possibly due to low-specificity interactions with the surface of the protein or restriction in the conformation of the glycans. In contrast, when solution-phase networking is avoided, enhancement due to binding of multiple branches of a glycan to multiple CRDs in the oligomeric forms of these receptors is minimal and binding of a receptor oligomer to multiple glycans on a single glycoprotein makes only a twofold contribution to overall affinity. Thus, in these cases, multivalent interactions of individual glycoproteins with individual receptor oligomers have a limited role in achieving high affinity. These findings, combined with considerations of membrane receptor geometry, are consistent with the idea that further enhancement of the binding to multivalent glycoprotein ligands requires interaction of multiple receptor oligomers with the ligands. PMID- 20004211 TI - Evolution of resistance to anti-cancer therapy during general dosing schedules. AB - Anti-cancer drugs targeted to specific oncogenic pathways have shown promising therapeutic results in the past few years; however, drug resistance remains an important obstacle for these therapies. Resistance to these drugs can emerge due to a variety of reasons including genetic or epigenetic changes which alter the binding site of the drug target, cellular metabolism or export mechanisms. Obtaining a better understanding of the evolution of resistant populations during therapy may enable the design of more effective therapeutic regimens which prevent or delay progression of disease due to resistance. In this paper, we use stochastic mathematical models to study the evolutionary dynamics of resistance under time-varying dosing schedules and pharmacokinetic effects. The populations of sensitive and resistant cells are modeled as multi-type non-homogeneous birth death processes in which the drug concentration affects the birth and death rates of both the sensitive and resistant cell populations in continuous time. This flexible model allows us to consider the effects of generalized treatment strategies as well as detailed pharmacokinetic phenomena such as drug elimination and accumulation over multiple doses. We develop estimates for the probability of developing resistance and moments of the size of the resistant cell population. With these estimates, we optimize treatment schedules over a subspace of tolerated schedules to minimize the risk of disease progression due to resistance as well as locate ideal schedules for controlling the population size of resistant clones in situations where resistance is inevitable. Our methodology can be used to describe dynamics of resistance arising due to a single (epi)genetic alteration in any tumor type. PMID- 20004212 TI - Effect of rapid human N-acetyltransferase 2 haplotype on DNA damage and mutagenesis induced by 2-amino-3-methylimidazo-[4,5-f]quinoline (IQ) and 2-amino 3,8-dimethylimidazo-[4,5-f]quinoxaline (MeIQx). AB - Heterocyclic amines such as 2-amino-3-methylimidazo-[4,5-f]quinoline (IQ) and 2 amino-3,8-dimethylimidazo-[4,5-f]quinoxaline (MeIQx) are dietary carcinogens generated when meats are cooked well-done. Bioactivation includes N-hydroxylation catalyzed by cytochrome P4501A2 (CYP1A2) followed by O-acetylation catalyzed by N acetyltransferase 2 (NAT2). Nucleotide excision repair-deficient Chinese hamster ovary (CHO) cells stably transfected with human CYP1A2 and either NAT2*4 (rapid acetylator) or NAT2*5B (slow acetylator) alleles were treated with IQ or MeIQx to examine the effect of NAT2 genetic polymorphism on IQ- or MeIQx-induced DNA adducts and mutagenesis. MeIQx and IQ both induced decreases in cell survival and significantly (p<0.001) greater number of endogenous hypoxanthine phosphoribosyl transferase (hprt) mutants in the CYP1A2/NAT2*4 than the CYP1A2/NAT2*5B cell line. IQ- and MeIQx-induced hprt mutant cDNAs were sequenced and over 85% of the mutations were single-base substitutions with the remainder exon deletions likely caused by splice-site mutations. For the single-base substitutions, over 85% were at G:C base pairs. Deoxyguanosine (dG)-C8-IQ and dG-C8-MeIQx adducts were significantly (p<0.001) greater in the CYP1A2/NAT2*4 than the CYP1A2/NAT2*5B cell line. DNA adduct levels correlated very highly with hprt mutants for both IQ and MeIQx. These results suggest substantially increased risk for IQ- and MeIQx induced DNA damage and mutagenesis in rapid NAT2 acetylators. PMID- 20004213 TI - Predicting the hepatocarcinogenic potential of alkenylbenzene flavoring agents using toxicogenomics and machine learning. AB - Identification of carcinogenic activity is the primary goal of the 2-year bioassay. The expense of these studies limits the number of chemicals that can be studied and therefore chemicals need to be prioritized based on a variety of parameters. We have developed an ensemble of support vector machine classification models based on male F344 rat liver gene expression following 2, 14 or 90 days of exposure to a collection of hepatocarcinogens (aflatoxin B1, 1 amino-2,4-dibromoanthraquinone, N-nitrosodimethylamine, methyleugenol) and non hepatocarcinogens (acetaminophen, ascorbic acid, tryptophan). Seven models were generated based on individual exposure durations (2, 14 or 90 days) or a combination of exposures (2+14, 2+90, 14+90 and 2+14+90 days). All sets of data, with the exception of one yielded models with 0% cross-validation error. Independent validation of the models was performed using expression data from the liver of rats exposed at 2 dose levels to a collection of alkenylbenzene flavoring agents. Depending on the model used and the exposure duration of the test data, independent validation error rates ranged from 47% to 10%. The variable with the most notable effect on independent validation accuracy was exposure duration of the alkenylbenzene test data. All models generally exhibited improved performance as the exposure duration of the alkenylbenzene data increased. The models differentiated between hepatocarcinogenic (estragole and safrole) and non-hepatocarcinogenic (anethole, eugenol and isoeugenol) alkenylbenzenes previously studied in a carcinogenicity bioassay. In the case of safrole the models correctly differentiated between carcinogenic and non carcinogenic dose levels. The models predict that two alkenylbenzenes not previously assessed in a carcinogenicity bioassay, myristicin and isosafrole, would be weakly hepatocarcinogenic if studied at a dose level of 2 mmol/kg bw/day for 2 years in male F344 rats; therefore suggesting that these chemicals should be a higher priority relative to other untested alkenylbenzenes for evaluation in the carcinogenicity bioassay. The results of the study indicate that gene expression-based predictive models are an effective tool for identifying hepatocarcinogens. Furthermore, we find that exposure duration is a critical variable in the success or failure of such an approach, particularly when evaluating chemicals with unknown carcinogenic potency. PMID- 20004214 TI - Reduction of negative afterimage duration in Parkinson's disease patients: a possible role for dopaminergic deficiency in the retinal Interplexiform cell layer. AB - Dopaminergic deficiency may affect Parkinson's disease patients (PD) in the central as well as the peripheral tissues. In the retina, the neuromodulatory role of the dopaminergic Interplexiform cell layer (IP) plays a major role in the retinal light adaptation and may account for the duration of the negative afterimage. Here we present results showing a significant reduction of negative afterimage duration in PD patients. This supports the hypothesis that the retinal dopaminergic system may be the main cause for the long duration of negative afterimage. We suggest that the observed reduction of afterimage duration is due to possible dopaminergic deficiency in patients with Parkinson's disease. PMID- 20004215 TI - Invasive group A streptococcal infections in children presenting to a paediatric intensive care unit in the North West of England. PMID- 20004216 TI - Antiviral prevention of sepsis induced cytomegalovirus reactivation in immunocompetent mice. AB - INTRODUCTION: Immunocompetent patients can reactivate latent cytomegalovirus (CMV) during critical illness and reactivation is associated with significantly worse outcomes. Prior to clinical trials in humans to prove causality, we sought to determine an optimal antiviral treatment strategy. METHODS: Mice latently infected with murine CMV (MCMV) received a septic reactivation trigger and were randomized to receive one of four ganciclovir regimens or saline. Lungs were evaluated for viral transcriptional reactivation and fibrosis after each regimen. Influences of ganciclovir on early sepsis-induced pulmonary inflammation and T cell activation were studied after sepsis induction. RESULTS: All ganciclovir regimens reduced measurable MCMV transcriptional reactivation, and 10mg/day for 7 or 21 days was most effective. Lower dose (5mg/kg/day) or delayed therapy was associated with significant breakthrough reactivation. Higher doses of ganciclovir given early were associated with the lowest incidence of pulmonary fibrosis, and delay of therapy for 1 week was associated with significantly worse pulmonary fibrosis. Although bacterial sepsis induced activation of MCMV-specific pulmonary T-cells, this activation was not influenced by ganciclovir. CONCLUSION: These results suggest that antiviral treatment trials in humans should use 10mg/kg/day ganciclovir administered as early as possible in at-risk patients to minimize reactivation events and associated pulmonary injury. PMID- 20004217 TI - HIV-1 non-B subtypes: High transmitted NNRTI-resistance in Spain and impaired genotypic resistance interpretation due to variability. AB - Genotypic resistance algorithms interpret drug-resistance mutations, but are mainly developed for HIV-1 subtype B, meanwhile non-B subtypes cause 90% of worldwide infections. They include clade-specific amino acid at drug-resistance positions different than subtype B. This study explores: (i) the variability at resistance-related positions in 128 non-B and 226 B sequences from 354 treatment naive patients diagnosed in Spain (1999-2007); (ii) the discordances between five resistance interpretation algorithms (ANRS, Stanford, Rega, Geno2pheno, RIS); and (iii) the reliability of five subtyping tools (Stanford, Geno2pheno, Rega, NCBI, EuResist) for each HIV-1 variant. Primary drug-resistance prevalence was 13.6%, although higher in non-B vs. B subtypes (18.7% vs. 10.6%), due to a twofold higher NNRTI-resistance prevalence (15.7% vs. 7.6%). Most secondary PI resistances, more frequent in non-B, were in fact clade-specific residues. Most sequences were interpreted as susceptible to all antiretrovirals by the five resistance algorithms, except for tipranavir by ANRS in non-B clades. Interalgorithm discordances were significantly higher in non-B variants for specific drugs. The agreement with phylogenetic analysis differed among subtyping tools testing non-B variants. We found a higher prevalence of NNRTI-resistance mutations in non-B subtypes. Certain algorithms overestimate the resistance in non-B subtypes due to natural patterns of mutations. Subtyping tools should be optimised for non-B variants. PMID- 20004218 TI - Early behavioural markers of disease in P301S tau transgenic mice. AB - The present study aimed at characterizing the early behavioural changes preceding the onset of disease in a transgenic mouse model of tauopathy. Mice transgenic for human P301S tau protein exhibit many characteristics of human tauopathies, including neurodegeneration, the formation of neuronal tau inclusions in the brain and the development of a pronounced motor phenotype by 5 months of age. In this study we analyzed the ultrasonic vocalization pattern in the first week of life and the response to nest-related olfactory stimuli on postnatal day 10 in both homozygous and heterozygous transgenic mice in comparison with the behavioural responses of C57BL/6J pups. Expression of human tau in the brain and spinal cord was evaluated by Western blot analysis. Homozygous P301S tau transgenic and C57BL/6J mice were also assessed for motor coordination in the Rotarod apparatus at 1, 2, 3 and 4 months of age, and spatial learning and spontaneous behaviour were evaluated at 2 months in the Morris water-maze and the open-field test. Results showed that P301S pups emitted more ultrasounds than C57BL/6J pups when separated from the mother. Two-month-old P301S mice displayed a hyperactive profile, as shown by increased swimming speed, enhanced locomotion and exploration of a novel object in the open field. However, the spatial task acquisition was unaltered. Decline of motor coordination capacities was evident only by the third month of age in the rotarod test. Behavioural abnormalities can be detected well before occurrence of neurodegeneration in this transgenic strain, thus representing precocious markers of pathology. PMID- 20004219 TI - The orbitofrontal cortex is not necessary for acquisition or remote recall of socially transmitted food preferences. AB - A role for prefrontal cortex has been proposed in systems consolidation of memory. The current study examined the effects of excitotoxic lesions of the orbitofrontal cortex (OFC) in rats on acquisition and remote recall of socially transmitted food preferences (STFP). Subjects received excitotoxic lesions of the OFC, and they were trained on two food preferences. They were tested 1h after the first training session to determine the effect of the lesion on acquisition. The following day, they were trained on a second preference and tested 10 days later to determine the effect of the lesion on remote recall. OFC lesions did not impair either STFP acquisition or remote recall in comparisons with sham-operated animals. In addition, a subset of animals underwent odor discrimination and reversal training. Consistent with previous reports, subjects with OFC lesions required more trials to reach criterion and made more errors during reversal training than did sham-operated animals. Taken together, the results of the present study indicate that the orbitofrontal cortex is not necessary for acquisition or systems consolidation of socially transmitted food preferences. PMID- 20004220 TI - An efficient DNA isolation protocol for filamentous cyanobacteria of the genus Arthrospira. AB - Thanks to their photosynthetic and nutritive properties, cyanobacteria of the Arthrospira genus are of interest as food supplements, as efficient oxygen producing life support system organisms for manned space flight, and for the production of biofuels. Despite these potential valuable applications, full genome sequences and genetic information in general on Arthrospira remain scarce. This is mainly due to the difficulty to extract sufficient high molecular weight nucleic acids from these filamentous cyanobacteria. In this article, an efficient and reproducible DNA extraction procedure for cyanobacteria of the genus Arthrospira was developed. The method is based on the combination of a soft mechanical lysis with enzymatic disruption of the cell wall. The comparison with other extraction protocols clearly indicates that this optimised method allows the recovery of a larger amount of DNA. Furthermore, the extracted DNA presents a high molecular weight, a reduced degradation and an excellent overall quality. It can be directly used for molecular biology purposes such as PCR, and clone library construction. PMID- 20004222 TI - The forthcoming applications of gold nanoparticles in drug and gene delivery systems. AB - The unique optical, chemical, and biological properties of gold nanoparticles have resulted in them becoming of clinical interest in several applications including drug and gene delivery. The attractive features of gold nanoparticles include their surface plasmon resonance, the controlled manner in which they interact with thiol groups, and their non-toxic nature. These attributes can be exploited to provide an effective and selective platform to obtain a targeted intracellular release of some substance. The use of gold nanoparticles can also increase the stability of the payload. Here we review recent advances in the use of gold nanoparticles in drug and gene delivery systems. The topics of surface modification, site-specificity and drugs and gene and gene delivery are discussed. PMID- 20004221 TI - Neurocognitive mechanisms of conceptual processing in healthy adults and patients with schizophrenia. AB - This overview outlines findings of cognitive and neurocognitive studies on comprehension of verbal, pictorial, and video stimuli in healthy participants and patients with schizophrenia. We present evidence for a distinction between two complementary neurocognitive streams of conceptual analysis during comprehension. In familiar situations, adequate understanding of events may be achieved by mapping the perceived information on the associative and similarity-based connections between concepts in semantic memory - a process reflected by an N400 waveform of event-related electrophysiological potentials (ERPs). However, in less conventional contexts, a more flexible mechanism may be needed. We suggest that this alternative processing stream, reflected by a P600 ERP waveform, may use discrete, rule-like goal-related requirements of real-world actions to comprehend relationships between perceived people, objects, and actions. This neurocognitive model of comprehension is used as a basis in discussing studies in schizophrenia. These studies suggest an imbalanced engagement of the two conceptual streams in schizophrenia, whereby patients may rely on the associative and similarity-based networks in semantic memory even when it would be more adaptive to recruit mechanisms that draw upon goal-related requirements. Finally, we consider the roles that these conceptual mechanisms may play in real-life behavior, and the consequences that their dysfunction may have for disorganized behavior and inability to plan actions to achieve behavioral goals in schizophrenia. PMID- 20004224 TI - Thirty years of the journal Appetite. The citation record. PMID- 20004223 TI - Self-controlled children stay leaner in the transition to adolescence. AB - In a prospective longitudinal study, we examined whether the personality trait of self-control protects against weight gain during the transition from childhood to adolescence. We obtained multi-method, multi-source measures of self-control from a socioeconomically and ethnically diverse sample of 105 fifth-grade students. Height and weight were recorded by the school nurse and used to calculate age- and gender-specific standardized body mass index (BMI) z-scores. Self-controlled fifth graders had lower BMI z-scores in eighth grade compared to their more impulsive peers, and this relationship remained significant when controlling for potential confounds, including gender, age, socioeconomic status, ethnicity, IQ, and happiness. Moreover, when controlling for the same covariates, self-control measured in fifth grade predicted decreases in BMI z-scores from fifth to eighth grade. These results suggest that more self-controlled children are protected from weight gain in the transition to adolescence. PMID- 20004225 TI - Copper accumulation by cultured astrocytes. AB - To study copper transport in brain astrocytes, we have used astrocyte-rich primary cultures as model system. Cells in these cultures contained a basal copper content of 1.1+/-0.4 nmol per mg protein. The cellular copper content increased strongly after application of copper chloride in a time and concentration-dependent manner. Analysis of the linear copper accumulation during the first 5 min of copper exposure revealed that cultured astrocytes accumulated copper with saturable kinetics with apparent K(M)- and V(max)-values of 9.4+/-1.8 microM and 0.76+/-0.13 nmol/(min x mg protein), respectively. In contrast, incubation of astrocytes with copper in the presence of ascorbate caused a linear increase of the copper accumulation rates for copper concentrations of up to 30 microM. In addition, copper accumulation was strongly inhibited by the presence of an excess of zinc or of various other divalent metal ions. The presence of mRNA and of immunoreactivity of the copper transport protein Ctr1 in astrocyte cultures suggests that Ctr1 contributes to the observed copper accumulation. However, since some characteristics of the observed copper accumulation are not consistent with Ctr1-mediated copper transport, additional Ctr1-independent mechanism(s) are likely to be involved in astrocytic copper accumulation. PMID- 20004226 TI - Genetically modified plants for non-food or non-feed purposes: straightforward screening for their appearance in food and feed. AB - Genetically modified (GM) plants aimed at producing food/feed are part of regular agriculture in many areas of the World. Commodity plants have also found application as bioreactors, designated non-food/non-feed GM (NFGM) plants, thereby making raw material for further refinement to industrial, diagnostic or pharmaceutical preparations. Many among them may pose health challenge to consumers or livestock animals, if occurring in food/feed. NFGM plants are typically released into the environment, but are grown under special oversight and any among several containment practices, none of which provide full protection against accidental dispersal. Adventitious admixture with food or feed can occur either through distributional mismanagement or as a consequence of gene flow to plant relatives. To facilitate NFGM surveillance we propose a new mandatory tagging of essentially all such plants, prior to cultivation or marketing in the European Union. The suggested tag--Plant-Made Industrial or Pharmaceutical Products Tag (PMIP-T)--is envisaged to occur as a transgenic silent DNA identifier in host plants and designed to enable technically simple identification and characterisation of any NFGM. Implementation of PMIP-T would permit inexpensive, reliable and high-throughput screening for NFGM specifically. The paper outlines key NFGM prospects and challenges as well as the PMIP-T concept. PMID- 20004227 TI - A topological substructural molecular design approach for predicting mutagenesis end-points of alpha, beta-unsaturated carbonyl compounds. AB - Chemically reactive, alpha, beta-unsaturated carbonyl compounds are common environmental pollutants able to produce a wide range of adverse effects, including, e.g. mutagenicity. This toxic property can often be related to chemical structure, in particular to specific molecular substructures or fragments (alerts), which can then be used in specialized software or expert systems for predictive purposes. In the past, there have been many attempts to predict the mutagenicity of alpha, beta-unsaturated carbonyl compounds through quantitative structure activity relationships (QSAR) but considering only one exclusive endpoint: the Ames test. Besides, even though those studies give a comprehensive understanding of the phenomenon, they do not provide substructural information that could be useful forward improving expert systems based on structural alerts (SAs). This work reports an evaluation of classification models to probe the mutagenic activity of alpha, beta-unsaturated carbonyl compounds over two endpoints--the Ames and mammalian cell gene mutation tests--based on linear discriminant analysis along with the topological Substructure molecular design (TOPS-MODE) approach. The obtained results showed the better ability of the TOPS-MODE approach in flagging structural alerts for the mutagenicity of these compounds compared to the expert system TOXTREE. Thus, the application of the present QSAR models can aid toxicologists in risk assessment and in prioritizing testing, as well as in the improvement of expert systems, such as the TOXTREE software, where SAs are implemented. PMID- 20004228 TI - Challenges of using pluripotent stem cells for safety assessments of substances. AB - Various European Union (EU) legislations request the use of in vitro tests for toxicological evaluations in order to increase the safety of consumer and patients but also to reduce the number vertebrates. The review provides a brief overview on EU legislations in place but without further interpretation. At present several ongoing EU projects address the need of developing predictive in vitro tests including projects assessing the potential of human embryonic stem cell (hESC) lines as basis for a range of toxicity tests. Tests based on human cells would avoid interspecies variations and as such predict more precisely adverse effects to the human body. However, the ethical situation on the use of toxicity tests based on hESCs is still under debate since no harmonization within Europe on the use of hESC lines has been achieved yet. A mutual acceptance of toxicity tests based on hESCs for regulatory applications is therefore challenging. Recent reports on the establishment of induced pluripotent stem cells (iPSC) are pointing to a way out of this dilemma, since these cells have apparently very similar characteristics as hESCs and could serve as basis for the development of toxicity tests. A careful scientific comparison between pluripotent cells of different origin is now needed in order to make final judgments. In any case, the development of reliable and relevant in vitro toxicity tests based on human pluripotent cells requires additional quality assessments of critical parameter that are also summarized within the review. PMID- 20004229 TI - Enzymatic degradation products from a marine polysaccharide YCP with different immunological activity and binding affinity to macrophages, hydrolyzed by alpha amylases from different origins. AB - YCP is a marine polysaccharide with anti-tumor and immune-modulating effects. This study evaluated the effect of enzymatic degradation of YCP by alpha-amylases from different origins on its immunological activity and binding ability to the macrophages. YCP was hydrolyzed by alpha-amylases isolated from Aspergillus oryzae, Bacillus licheniformis, Barley malt, and Porcine pancreas respectively, then four fragments with unique molecular weight (termed: YCP-Ao, YCP-Bl, YCP-Bm, and YCP-Pp, respectively) were obtained. The four fragments showed different immunological activity and the ability to bind to macrophages. Among them, YCP-Ao possessed almost equivalent immunological activity compared to the original YCP, while such properties were not retained in YCP-Bl. Our further study showed that YCP-Ao prevented YCP from binding to macrophages. In conclusion, YCP-Ao and YCP might have similar active regions. PMID- 20004231 TI - Arousal response to hypoxia in newborns: Insights from animal models. AB - In newborns, the inability to initiate an arousal response to hypoxia is associated with apnea of prematurity, sudden infant death syndrome, and rare genetic disorders of respiratory control. Despite intensive research, the mechanisms of this response are poorly understood. This paper provides an overview of studies investigating the arousal response to hypoxia, with special emphasis on newborn mouse models. Mutant mouse models can provide valuable information regarding the pathogenesis of genetically determined disorders affecting arousal response to hypoxia, although data remain sparse. In mice, the arousal response to hypoxia emerges immediately after birth, when the ventilatory response to hypoxia is still immature. Habituation of the arousal response occurs after repeated hypoxic episodes. Newborn mice can learn to associate novel odors to hypoxia and respond to those odors by producing alerting responses, suggesting that the arousal response to hypoxia may be shaped by learning processes. PMID- 20004232 TI - Global architecture of metabolite distributions across species and its formation mechanisms. AB - Living organisms produce metabolites of many types via metabolic reactions. Especially, flavonoids, a type of secondary metabolite, of plant species are interesting examples. Since plant species are believed to have specific flavonoids that were formed response to diverse environments, elucidation of the design principles of metabolite distributions across plant species is important for understanding metabolite diversity and plant evolution. In our previous study, we found heterogeneous connectivity in metabolite distribution, and proposed a simple model to explain the possible origin of heterogeneous connectivity. In this paper, we determine additional structural properties of metabolite distribution among families that are analogous analogy with plant animal mutualistic networks: nested structure and modular structure. An earlier model representative of these structural properties in bipartite relationships was established on the basis of the traits of elements and external factors. However, we found that the architecture of metabolite distribution could be explained by simple evolutionary processes without trait-based mechanisms by comparing our model and the earlier model. Our model provided a better qualitative and quantitative prediction of nested and modular structures in addition to heterogeneous connectivity. This finding implies an alternative possible origin of these structural properties, and suggests the mechanisms underlying establishment metabolite distributions across plant species are simpler than expected. PMID- 20004230 TI - Emergence and re-emergence of viral diseases of the central nervous system. AB - Neurologic disease is a major cause of disability in resource-poor countries and a substantial portion of this disease is due to infections of the CNS. A wide variety of emerging and re-emerging viruses contribute to this disease burden. New emerging infections are commonly due to RNA viruses that have expanded their geographic range, spread from animal reservoirs or acquired new neurovirulence properties. Mosquito-borne viruses with expanding ranges include West Nile virus, Japanese encephalitis virus and Chikungunya virus. Zoonotic viruses that have recently crossed into humans to cause neurologic disease include the bat henipaviruses Nipah and Hendra, as well as the primate-derived human immunodeficiency virus. Viruses adapt to new hosts, or to cause more severe disease, by changing their genomes through reassortment (e.g. influenza virus), mutation (essentially all RNA viruses) and recombination (e.g. vaccine strains of poliovirus). Viruses that appear to have recently become more neurovirulent include West Nile virus, enterovirus 71 and possibly Chikungunya virus. In addition to these newer challenges, rabies, polio and measles all remain important causes of neurologic disease despite good vaccines and global efforts toward control. Control of human rabies depends on elimination of rabies in domestic dogs through regular vaccination. Poliovirus eradication is challenged by the ability of the live attenuated vaccine strains to revert to virulence during the prolonged period of gastrointestinal replication. Measles elimination depends on delivery of two doses of live virus vaccine to a high enough proportion of the population to maintain herd immunity for this highly infectious virus. PMID- 20004233 TI - Investigation of gammaE-crystallin target protein binding to bovine lens alpha crystallin by small-angle neutron scattering. AB - alpha-Crystallin, one of the main constituent proteins in the crystalline lens, is an important molecular chaperone both within and outside the lens. Presently, the structural relationship between alpha-crystallin and its target proteins during chaperone action is poorly understood. It has been hypothesised that target proteins bind within a central cavity. Small-angle neutron-scattering (SANS) experiments in conjunction with isotopic substitution were undertaken to investigate the interaction of a target lens protein (gammaE-crystallin) with alpha-crystallin (alpha(H)) and to measure the radius of gyration (Rg) of the proteins and their binary complexes in solution under thermal stress. The size of the alpha(H) in D(2)O incubated at 65 degrees C increased from 69+/-3 to 81+/-5 A over 40 min, in good agreement with previously published small-angle X-ray scattering (SAXS) and SANS measurements. Deuterated gammaE-crystallin in H(2)O buffer (gammaE(D)/H(2)O) and hydrogenous gammaE-crystallin in D(2)O buffer (gammaE(H)/D(2)O) free in solution were of insufficient size and/or too dilute to provide any measurable scattering over the angular range used, which was selected primarily to investigate gammaE:alpha(H) complexes. The evolution of the aggregation size/shape as an indicator of alpha(H) chaperone action was monitored by recording the neutron scattering in different H:D solvent contrasts under thermally stressed conditions (65 degrees C) for binary mixtures of alpha(H), gammaE(H), and gammaE(D). It was found that Rg(alpha(H):gammaE(D)/D(2)O)>Rg(alpha(H):gammaE(H)/D(2)O)>Rg(alpha(H)/D(2)O) and that Rg(alpha(H):gammaE(H)/D(2)O) approximately Rg(alpha(H)/D(2)O). The relative sizes observed for the complexes weighted by the respective scattering powers of the various components imply that gammaE-crystallin binds in a central cavity of the alpha-crystallin oligomer, during chaperone action. PMID- 20004234 TI - Pre-treatment of Syndrex protects mice from becoming diabetic after streptozotocin injection. AB - Diabetes mellitus is a metabolic disorder characterized by hyperglycemia due to either insufficiency of insulin or inability of cells to respond to insulin. Many clinical and experimental evidence have suggested the strong association between hyperglycemia, oxidative stress and diabetic complications. Therefore, the antidiabetic drugs with antioxidant potential would have a higher therapeutic value. To check its antidiabetic and antioxidant properties in vivo, experiments were done wherein mice were fed with Syndrex in different schedules and/or made diabetic by intraperitoneal injection of streptozotocin. Animals fed with Syndrex prior to the induction of diabetes by streptozotocin injection showed resistance to an increase in blood glucose levels. This treatment increased the activities of antioxidant enzymes namely, catalase, glutathione reductase and superoxide dismutase and reduced serum triglyceride and cholesterol levels as compared to those found in uncontrolled diabetic mice. Among the three different schedules used for Syndrex treatment, the best effect was seen in the case of mice pretreated with Syndrex prior to STZ injection. In our opinion, Syndrex given along with insulin may reduce the amount of insulin dose required and because of its strong antioxidant activity would certainly help to reduce the development of diabetic complications. PMID- 20004235 TI - Characterization of luciferases and its paralogue in the Panamanian luminous click beetle Pyrophorus angustus: a click beetle luciferase lacks the fatty acyl CoA synthetic activity. AB - Two luciferase genes (dPaLuc and vPaLuc) and one paralogue of luciferase (PaLL) were isolated from the Panamanian luminous click beetle, Pyrophorus angustus (Elateridae, Pyrophorinae). The transcripts of dPaLuc and vPaLuc were predominantly detected in the body parts with dorsal photophore and ventral photophore, respectively, and the transcript of PaLL was detected in both parts. The gene products of dPaLuc and vPaLuc possessed luminescence activity with firefly luciferin (lambda(max)=536 and 566 nm, respectively) but did not show significant activity of fatty acyl-CoA synthesis. On the other hand, the gene product of PaLL had fatty acyl-CoA synthetic activity with very weak luminescence activity. The catalytic properties of click beetle luciferase are different from our previous results that firefly luciferase has both luminescence activity and fatty acyl-CoA synthetic activity. These results suggested that the ancestral fatty acyl-CoA synthetase in the Pyrophorinae lineage has undergone gene duplication event, followed by specialization of one copy in luciferase. Subsequently, the luciferase was duplicated again and the two copies diverged in their luminescent color and expression pattern. PMID- 20004236 TI - Over-expression of a protein kinase gene enhances the defense of tobacco against Rhizoctonia solani. AB - To identify Nicotiana tabacum genes involved in resistance and susceptibility to Rhizoctonia solani, suppression subtractive hybridization was used to generate a cDNA library from transcripts that are differentially expressed during a compatible and incompatible interaction. This allowed the isolation of a protein kinase cDNA that was down-regulated during a compatible and up-regulated during an incompatible interaction. Quantitative RT-PCR analysis of this gene confirmed the differential expression patterns between the compatible and incompatible interactions. Over-expression of this gene in tobacco enhanced the resistance to damping-off produced by an aggressive R. solani strain. Furthermore, silencing of this protein kinase gene reduced the resistance to a non-aggressive R. solani strain. A set of reported tobacco-resistant genes were also evaluated in tobacco plants over-expressing and silencing the protein kinase cDNA. Several genes previously associated with resistance in tobacco, like manganese superoxide dismutase, Hsr203J, chitinases and phenylalanine ammonia-lyase, were up-regulated in tobacco plants over-expressing the protein kinase cDNA. Potentially, the protein kinase gene could be used to engineer resistance to R. solani in tobacco cultivars susceptible to this important pathogen. PMID- 20004237 TI - Why Surinamese migrants in the Netherlands continue to use medicinal herbs from their home country. AB - AIM OF THE STUDY: When people migrate, they tend to bring along their medicinal plants. In order to improve migrant health, we need information on their traditional health beliefs and practices. This paper investigates medicinal plant use among Surinamese migrants in the Netherlands. MATERIALS AND METHODS: Data from 210 semi-structured interviews among 1st and 2nd generation Surinamese migrants were analysed to determine which medicinal plants were used, for what purposes, which demographic, socio-economic or psycho-social factors play a role in the choice for traditional medicine and to clarify people's personal motives to use herbs. Variables associated with medicinal plant use were identified by using the Pearson gamma2 test and the two-sample t-test. After selecting significant variables by means of bivariate analyses, multinomial logistic regression with stepwise forward selection was used to assess whether medicinal plant use could be explained by a combination of these variables. RESULTS: More than 75% of the respondents used herbal medicine, and 66% did so in the past year. Herbs were more frequently employed for health promotion (39%) than for disease prevention or cure (both 27%). Almost half of the respondents who had been ill the last year had used herbal medicine. More than 140 herb species were mentioned during the interviews. Plant use was often related to certain culture bound health beliefs. Spiritual baths were the most popular traditional practice, followed by genital steam baths, bitter tonics, and the consumption of bitter vegetables. Afro-Surinamers more frequently used herbal medicine than Hindustani. The WINTI belief strongly influenced plant use, as well as the occurrence of an illness in the past year, and frequent visits to Suriname. Age, gender, income and education had no significant effect on the use of traditional medicine. Surinamers stated that they used medicinal herbs because they grew up with them; herbs were more effective and had fewer side effects than conventional therapies. CONCLUSIONS: As long as certain culture-bound beliefs and health concepts remain prevalent among Surinamese migrants, and ties with their home country remain strong, they will continue using medicinal herbs from their country of origin. More research is needed on the health effects of frequently used medicinal plants by migrants in the Netherlands. PMID- 20004238 TI - Does mtDNA nucleoid organization impact aging? AB - Somatic cells in tissue culture package several copies of mitochondrial DNA (mtDNA) in aggregates known as nucleoids that appear to be remarkably stable. The clustering of multiple mtDNA genomes in a single nucleoid complex may promote the progressive age-related accumulation of deletion and point mutations in mtDNA in many somatic tissues, particularly in post-mitotic cells. In contrast, oocytes appear to have the ability to select against deleterious mutations in mtDNA, at least in mice. This fundamental difference suggests that oocytes may be better able to detect and remove defective mtDNA genomes than somatic cells, possibly due in part to the simpler organization of the mtDNA in smaller nucleoids. These observations suggest the hypothesis that a complex nucleoid structure containing several mtDNA molecules may impair the ability of the cell to select against deleterious mtDNA mutations, thereby contributing to age-related mitochondrial dysfunction. PMID- 20004239 TI - Increased expression of endothelial iNOS accounts for hyporesponsiveness of pulmonary artery to vasoconstrictors after paraquat poisoning. AB - Paraquat is a toxic herbicide that induces severe acute lung injury (ALI) and pulmonary hypertension in humans. Although vascular disorders are present and contribute to increased mortality in ALI patients, there is little data available on vascular responsiveness after toxic exposure to paraquat. We aimed to evaluate the vascular response of isolated pulmonary arteries from rats treated with a dose of paraquat that induces ALI. Paraquat treatment did not modify the relaxant response of pulmonary artery to acetylcholine, but greatly reduced phenylephrine induced contraction. Removal of the endothelium, inhibition of nitric oxide synthase (NOS) with L-NAME or selective inhibition of inducible NOS (iNOS) with L NIL, restored contraction of vessels from paraquat poisoned rats to the same level as those not exposed to paraquat. The basal production of NO and expression of iNOS were increased in endothelium-intact but not in endothelium-denuded vessels from paraquat-poisoned rats. Expression of endothelial NOS was not modified. Our findings suggest that paraquat poisoning increases endothelial iNOS expression and basal NO production decreasing responsiveness of pulmonary artery to vasoconstrictors. Thus, our results do not support the hypothesis that pulmonary hypertension in paraquat-induced ALI is mediated by a reduction in endothelial NO production or increased contractility of pulmonary artery. PMID- 20004240 TI - Characterization of the rabbit agouti signaling protein (ASIP) gene: transcripts and phylogenetic analyses and identification of the causative mutation of the nonagouti black coat colour. AB - The agouti locus encodes the agouti signalling protein (ASIP) which is involved in determining the switch from eumelanin to pheomelanin synthesis in melanocytes. In the domestic rabbit (Oryctolagus cuniculus) early studies indicated three alleles at this locus: A, light-bellied agouti (wild type); a(t), black and tan; a, black nonagouti. We characterized the rabbit ASIP gene and identified the causative mutation (an insertion in exon 2) of the black nonagouti allele whose frequency was evaluated in 31 breeds. Phylogenetic analysis of ASIP sequences from Oryctolagus and 9 other species of the family Leporidae placed Oryctolagus as sister species to Pentalagus and Bunolagus. Transcription analysis in wild type agouti rabbits revealed the presence of two major transcripts with different 5'-untranslated regions having ventral or dorsal skin specific expression. ASIP gene transcripts were also detected in all examined rabbit tissues distinguishing the rabbit expression pattern from what was observed in wild type mice. PMID- 20004242 TI - Differential activation of p38MAPK isoforms by MKK6 and MKK3. AB - All four members of the mammalian p38 mitogen-activated protein kinase (MAPK) family (p38alpha, p38beta, p38gamma and p38delta) are activated by dual phosphorylation in the TGY motif in the activation loop. This phosphorylation is mediated by three kinases, MKK3, MKK6 and MKK4, at least in vitro. The role of these MKK in the activation of p38alpha has been demonstrated in studies using fibroblasts that lack MKK3 and/or MKK6. Nonetheless, the physiological upstream activators of the other p38MAPK isoforms have not yet been reported using MKK knockout cells. In this study, we examined p38beta, gamma and delta activation by MKK3 and MKK6, in cells lacking MKK3, MKK6 or both. We show that MKK3 and MKK6 are both essential for the activation of p38gamma and p38beta induced by environmental stress, whereas MKK6 is the major p38gamma activator in response to TNFalpha. In contrast, p38delta activation by ultraviolet radiation, hyperosmotic shock, anisomycin or by TNFalpha is mediated by MKK3. Moreover, in response to osmotic stress, MKK3 and MKK6 are crucial in regulating the phosphorylation of the p38gamma substrate hDlg and its activity as scaffold protein. These data indicate that activation of distinct p38MAPK isoforms is regulated by the selective and synchronized action of two kinases, MKK3 and MKK6, in response to cell stress. PMID- 20004241 TI - Oxidative stress from diverse developmental neurotoxicants: antioxidants protect against lipid peroxidation without preventing cell loss. AB - Oxidative stress has been hypothesized to provide a mechanism by which apparently unrelated chemicals can nevertheless produce similar developmental neurotoxic outcomes. We used differentiating PC12 cells to compare the effects of agents from four different classes and then to evaluate antioxidant amelioration: fipronil, perfluorooctanesulfonamide (PFOSA), dieldrin and chlorpyrifos. The rank order for lipid peroxidation corresponded to the ability to evoke cell loss: fipronil>PFOSA>dieldrin>chlorpyrifos. The same sequence was found for an index of cell enlargement (protein/DNA ratio) but the effects on neurite outgrowth (membrane/total protein) diverged, with fipronil producing a decrease and PFOSA an increase. Cotreatment with antioxidants reduced (ascorbate) or eliminated (Vitamin E) lipid peroxidation caused by each of the agents but failed to protect against cell loss, with the sole exception of chlorpyrifos, for which we earlier showed partial protection by Vitamin E; addition of higher NGF concentrations protected neither against oxidative stress nor cell loss. Despite the failure to prevent cell loss, ascorbate protected the cells from the effects of PFOSA on neuritic outgrowth; NGF, and to a lesser extent, ascorbate, offset the effects of fipronil on both cell enlargement and neuritogenesis. At the same time, the ameliorant treatments also worsened some of the other toxicant effects. Our results point out the problems in concluding that, just because a neurotoxicant produces oxidative stress, antioxidant therapy will be effective in preventing damage. Instead, additional mechanisms for each agent may provide alternative routes to neurotoxicity, or may be additive or synergistic with oxidative stress. PMID- 20004243 TI - A single-point mutation in FGFR2 affects cell cycle and Tgfbeta signalling in osteoblasts. AB - Fgf and Tgfbeta are key regulators of bone development. It is not known, however, whether there is a relationship between defective Fgf signalling, resulting in a premature cranial suture fusion, and Tgfbeta signalling. We used mouse calvaria osteoblasts carrying a mutation (hFGFR2-C278F) associated with Crouzon and Pfeiffer syndromes to investigate effects of this mutation on cell growth and possible mechanisms underlying it. Mutated osteoblasts displayed reduced S-phase, increased apoptosis and increased differentiation. As Tgfbeta signalling appeared to be required in an autocrine/paracrine manner for osteoblast proliferation, we tested the hypothesis that reduced growth might be due, at least in part, to an altered balance between FGF and Tgfbeta signalling. Tgfbeta expression was indeed decreased in mutated osteoblasts, as compared to osteoblasts carrying the wild type hFGFR2. Treatment with Tgfbeta, however, neither increased proliferation in mutated osteoblasts, unlike in controls, nor rescued proliferation in control osteoblasts treated with an Erk1/2 inhibitor. Significantly, Erk2, that is important for proliferation, was reduced relatively to Erk1 in mutated cells. Altogether this study suggests that the hFGFR2-C278F mutation affects the osteoblast ability to respond to Tgfbeta stimulation via the Erk pathway and that the overall effect of the mutation is a loss of function. PMID- 20004244 TI - Recent advances in the manipulation of murine gene expression and its utility for the study of human neurological disease. AB - Transgenic mouse models have vastly contributed to our knowledge of the genetic and molecular pathways underlying the pathogenesis of neurological disorders that affect millions of people worldwide. Not only have they allowed the generation of disease models mimicking the human pathological state but they have also permitted the exploration of the pathological role of specific genes through the generation of knock-out and knock-in models. Classical constitutive transgenic mice have several limitations however, due to behavioral adaptation process occurring and conditional mouse models are time-consuming and often lack extensive spatial or temporal control of gene manipulation. These limitations could be overcome by means of innovative methods that are now available such as RNAi, viral vectors and large cloning DNA vectors. These tools have been extensively used for the generation of mouse models and are characterized by the superior control of transgene expression that has been proven invaluable in the assessment of novel treatments for neurological diseases and to further investigate the molecular processes underlying the etiopathology of neurological disorders. Furthermore, in association with classical transgenic mouse models, they have allowed the validation of innovative therapeutic strategies for the treatment of human neurological disorders. This review describes how these tools have overcome the limitations of classical transgenic mouse models and how they have been of value for the study of human neurological diseases. PMID- 20004245 TI - Tau-knockout mice show reduced GSK3-induced hippocampal degeneration and learning deficits. AB - It has been proposed that deregulation of neuronal glycogen synthase kinase 3 (GSK3) activity may be a key feature in Alzheimer disease pathogenesis. We have previously generated transgenic mice that overexpress GSK3beta in forebrain regions including dentate gyrus (DG), a region involved in learning and memory acquisition. We have found that GSK3 overexpression results in DG degeneration. To test whether tau protein modified by GSK3 plays a role in that neurodegeneration, we have brought GSK3 overexpressing mice to a tau knockout background. Our results indicate that the toxic effect of GSK3 overexpression is milder and slower in the absence of tau. Thus, we suggest that the hyperphosphorylated tau mediates, at least in part, the pathology observed in the brain of GSK3 overexpressing mice. PMID- 20004246 TI - Antimicrobial peptides (AMP) with antiviral activity against fish nodavirus. AB - Nervous necrosis virus (NNV) is classified as betanodavirus of Nodaviridae, and has caused mass mortality of numerous marine fish species at larval stage. Antimicrobial peptides (AMPs) play an important role of innate immunity either against bacterial pathogens or viruses. Up to date, little is known if any AMP could effectively inhibit fish nodaviruses and its mechanism. In this study, the antiviral activities of three antimicrobial peptides (AMPs) against grouper NNV (GNNV) were screened in the fish cell line. Two of the three AMPs, tilapia hepcidin 1-5 (TH 1-5) and cyclic shrimp anti-lipopolysaccharide factor (cSALF), were able to agglutinate purified NNV particles into clump, and the clumps were further confirmed to be viral proteins by TEM and Western blot. The NNV solution, separately pre-mixed with AMP (TH 1-5 or cSALF) or deionized-distilled water for 1 h, was used to infect GF-1 cells, and the levels of capsid protein in the GNNV AMP-infected cells at 1 h post infection were much lower than that in the GNNV H(2)O-infected cells, indicating that only a small portion of viral particles in the GNNV-AMP mixture could successfully infected the cells. Treatment of cBB cells with TH 1-5 and cSALF did not induce Mx gene expression; however, grouper epinecidin-1 (CP643-1) could induce the expression of Mx in the pre-treated cBB cells. This study revealed three AMPs with anti-NNV activity through two different mechanisms, and shed light on the future application in aquaculture. PMID- 20004247 TI - Reading the mind's eye: decoding category information during mental imagery. AB - Category information for visually presented objects can be read out from multi voxel patterns of fMRI activity in ventral-temporal cortex. What is the nature and reliability of these patterns in the absence of any bottom-up visual input, for example, during visual imagery? Here, we first ask how well category information can be decoded for imagined objects and then compare the representations evoked during imagery and actual viewing. In an fMRI study, four object categories (food, tools, faces, buildings) were either visually presented to subjects, or imagined by them. Using pattern classification techniques, we could reliably decode category information (including for non-special categories, i.e., food and tools) from ventral-temporal cortex in both conditions, but only during actual viewing from retinotopic areas. Interestingly, in temporal cortex when the classifier was trained on the viewed condition and tested on the imagery condition, or vice versa, classification performance was comparable to within the imagery condition. The above results held even when we did not use information in the specialized category-selective areas. Thus, the patterns of representation during imagery and actual viewing are in fact surprisingly similar to each other. Consistent with this observation, the maps of "diagnostic voxels" (i.e., the classifier weights) for the perception and imagery classifiers were more similar in ventral-temporal cortex than in retinotopic cortex. These results suggest that in the absence of any bottom-up input, cortical back projections can selectively re-activate specific patterns of neural activity. PMID- 20004248 TI - Effect of hemodynamic variability on Granger causality analysis of fMRI. AB - In this work, we investigated the effect of the regional variability of the hemodynamic response on the sensitivity of Granger causality (GC) analysis of functional magnetic resonance imaging (fMRI) data to neuronal causal influences. We simulated fMRI data by convolving a standard canonical hemodynamic response function (HRF) with local field potentials (LFPs) acquired from the macaque cortex and manipulated the causal influence and neuronal delays between the LFPs, the hemodynamic delays between the HRFs, the signal-to-noise ratio (SNR), and the sampling period (TR) to assess the effect of each of these factors on the detectability of the neuronal delays from GC analysis of fMRI. In our first bivariate implementation, we assumed the worst-case scenario of the hemodynamic delay being at the empirical upper limit of its normal physiological range and opposing the direction of neuronal delay. We found that, in the absence of HRF confounds, even tens of milliseconds of neuronal delays can be inferred from fMRI. However, in the presence of HRF delays which opposed neuronal delays, the minimum detectable neuronal delay was hundreds of milliseconds. In our second multivariate simulation, we mimicked the real situation more closely by using a multivariate network of four time series and assumed the hemodynamic and neuronal delays to be unknown and drawn from a uniform random distribution. The resulting accuracy of detecting the correct multivariate network from fMRI was well above chance and was up to 90% with faster sampling. Generically, under all conditions, faster sampling and low measurement noise improved the sensitivity of GC analysis of fMRI data to neuronal causality. PMID- 20004249 TI - The neural substrates of impaired finger tapping regularity after stroke. AB - Not only finger tapping speed, but also tapping regularity can be impaired after stroke, contributing to reduced dexterity. The neural substrates of impaired tapping regularity after stroke are unknown. Previous work suggests damage to the dorsal premotor cortex (PMd) and prefrontal cortex (PFCx) affects externally-cued hand movement. We tested the hypothesis that these two areas are involved in impaired post-stroke tapping regularity. In 19 right-handed patients (15 men/4 women; age 45-80 years; purely subcortical in 16) partially to fully recovered from hemiparetic stroke, tri-axial accelerometric quantitative assessment of tapping regularity and BOLD fMRI were obtained during fixed-rate auditory-cued index-thumb tapping, in a single session 10-230 days after stroke. A strong random-effect correlation between tapping regularity index and fMRI signal was found in contralesional PMd such that the worse the regularity the stronger the activation. A significant correlation in the opposite direction was also present within contralesional PFCx. Both correlations were maintained if maximal index tapping speed, degree of paresis and time since stroke were added as potential confounds. Thus, the contralesional PMd and PFCx appear to be involved in the impaired ability of stroke patients to fingertap in pace with external cues. The findings for PMd are consistent with repetitive TMS investigations in stroke suggesting a role for this area in affected-hand movement timing. The inverse relationship with tapping regularity observed for the PFCx and the PMd suggests these two anatomically-connected areas negatively co-operate. These findings have implications for understanding the disruption and reorganization of the motor systems after stroke. PMID- 20004250 TI - Functional imaging of sympathetic activation during mental stress. AB - Activation of the sympathetic nervous system (SNS) is essential in adapting to environmental stressors and in maintaining homeostasis. This reaction can also turn into maladaptation, associated with a wide spectrum of stress-related diseases. Up to now, the cortical mechanisms of sympathetic activation in acute mental stress have not been sufficiently characterized. We therefore investigated cerebral activation applying functional magnetic resonance imaging (fMRI) during performance of a mental stress task with graded levels of difficulty, i.e. four versions of a Stroop task (Colour Word Interference Test, CWT) in healthy subjects. To analyze stress-associated sympathetic activation, skin conductance and heart rate were continuously recorded. The results show that sympathetic activation through mental stress is associated with distinct cerebral regions being immediately involved in task performance (visual, motor, and premotor areas). Other activated regions (right insula, dorsolateral superior frontal gyrus, cerebellar regions) are unrelated to task performance. These latter regions have previously been considered to be involved in mediating different stress responses. The results might furthermore serve as a basis for future investigations of the connection between these cortical regions in the generation of stress-related diseases. PMID- 20004251 TI - Perceiving the writing sequence of Chinese characters: an ERP investigation. AB - The neural dynamics in perceiving well-learned sequences and its modulation by task demand were investigated in this study in which participants were asked to observe stroke-by-stroke display of Chinese characters composed of two radicals while their brain activity was monitored with the event-related potential (ERP) technique. Experiment 1 used an accuracy judgment task that would draw participants' attention to the violation of the writing sequence whereas Experiment 2 required participants to judge the completion of the display and thus the more automatic aspects of sequence processing could be revealed. In Experiment 1, the within-radical boundary reversal produced bilateral posterior N2 enhancement and the cross-boundary reversal elicited a left N2 effect and right posterior N2 reduction on the critical stroke. Both types of reversal elicited P3 effects on the critical stroke and sustained negativity effects on the following stroke, with the size being larger for the cross-boundary reversal. In Experiment 2, in addition to the P3 effects, the within-boundary reversal elicited a left posterior N2 effect and the cross-boundary reversal elicited right posterior N2 reduction on the critical stroke. Moreover, on the following stroke, the cross-boundary reversal elicited a right N2 effect and both types of reversal elicited sustained positivity effects. These findings demonstrate that native Chinese readers use their sequential knowledge to predict upcoming strokes in perceiving the writing of characters and to construct appropriate representations for the action sequence regardless of whether such predictions and constructions are required by the task. PMID- 20004253 TI - A comparison of permutation and parametric testing for between group effective connectivity differences using DCM. AB - Effective connectivity is becoming an increasingly popular technique for obtaining additional information from functional magnetic resonance imaging (fMRI) cognitive activation studies. It is potentially important for investigating psychiatric illnesses, which are thought to depend on disrupted connections and in observing the action of psychoactive drugs used to treat these disorders. If researchers are to apply these techniques confidently then it is important to establish the level of power that is available in an experiment. This study compares the level of power available when applying effective connectivity to test for differences between groups using parametric tests and permutation testing. Permutation testing has previously been shown to have superior sensitivity to parametric tests in fMRI studies. As an illustrative example, both the parametric t-test and equivalent permutation test were applied to a comparison between healthy controls and remitted depressed volunteers performing an emotional face processing task. Permutation testing was found to provide superior power compared with the nonparametric equivalent. PMID- 20004252 TI - Abnormalities in neural processing of emotional stimuli in Williams syndrome vary according to social vs. non-social content. AB - Williams syndrome (WS) is a rare genetic disorder caused by the deletion of approximately 25 genes on chromosome 7q11.23 and is characterized by both hypersociability and increases in specific phobia and anticipatory anxiety regarding non-social entities or circumstances. Alterations in amygdala reactivity and prefrontal regulation consistent with the observed behavioral pattern of social versus non-social abnormalities have been previously demonstrated in individuals with WS (Meyer-Lindenberg et al., 2005). However, in that study, the social stimulus (faces) matching task was more difficult than the non-social scene (IAPS stimuli) matching task, making it impossible to disambiguate the relative contributions of task difficulty and stimulus type (social versus non-social). In the present study, we examined the performance of the same group of participants with WS and normal IQs during a more cognitively demanding task using the same scene stimuli as in the prior study. Confirming previous findings, the results indicated (a) a differential response of prefrontal regions as a function of task difficulty and (b) a persistently increased activation of the amygdala to non-social scenes by individuals with WS regardless of cognitive load. These data provide further evidence of disruption in amygdala-prefrontal circuitry in individuals with WS. PMID- 20004254 TI - Association between striatal and extrastriatal dopamine D2-receptor binding and social desirability. AB - Research on the biological underpinnings of personality can provide leads to the pathophysiology of psychiatric disorders. In particular, interpersonal aspects of behavior are a common problem during the course of psychiatric illness. Animal research has demonstrated a role for the dopamine system in social behaviour, and recent molecular imaging studies have shown a negative correlation between dopamine D2-receptor binding in the striatum and social desirability. The emotional and cognitive aspects of social behavior suggest involvement of brain regions outside of the striatum, such as limbic structures. The aim of the present study was to explore associations between the personality trait social desirability and dopamine D2-receptor binding in both striatal and extrastriatal brain regions. We examined 16 control subjects with Positron Emission Tomography and the radioligands [(11)C]raclopride and [(11)C]FLB 457, in relation to social desirability in the inventory Swedish universities Scales of Personality. [(11)C]raclopride D2-receptor binding in the striatum showed negative correlations to social desirability scores, corroborating previous findings. Furthermore, a correlation of a higher statistical significance was demonstrated for [(11)C]FLB 457 binding in the hippocampal-amygdala complex. A separate analysis of social desirability items in relation to a model of interpersonal behaviour revealed that the associations were driven by items reflecting high submissiveness and high affiliation. Taken together with previous evidence on D2 receptor binding and social behaviour, a role for dopaminergic neurotransmission in regulating displays of dominance vs. submissive behaviour is proposed. PMID- 20004255 TI - A novel approach for enhancing the signal-to-noise ratio and detecting automatically event-related potentials (ERPs) in single trials. AB - Brief radiant laser pulses can be used to activate cutaneous Adelta and C nociceptors selectively and elicit a number of transient brain responses in the ongoing EEG (N1, N2 and P2 waves of laser-evoked brain potentials, LEPs). Despite its physiological and clinical relevance, the early-latency N1 wave of LEPs is often difficult to measure reliably, because of its small signal-to-noise ratio (SNR), thus producing unavoidable biases in the interpretation of the results. Here, we aimed to develop a method to enhance the SNR of the N1 wave and measure its peak latency and amplitude in both average and single-trial waveforms. We obtained four main findings. First, we suggest that the N1 wave can be better detected using a central-frontal montage (Cc-Fz), as compared to the recommended temporal-frontal montage (Tc-Fz). Second, we show that the N1 wave is optimally detected when the neural activities underlying the N2 wave, which interfere with the scalp expression of the N1 wave, are preliminary isolated and removed using independent component analysis (ICA). Third, we show that after these N2-related activities are removed, the SNR of the N1 wave can be further enhanced using a novel approach based on wavelet filtering. Fourth, we provide quantitative evidence that a multiple linear regression approach can be applied to these filtered waveforms to obtain an automatic, reliable and unbiased estimate of the peak latency and amplitude of the N1 wave, both in average and single-trial waveforms. PMID- 20004257 TI - A new allele (eNOS4e) in the intron 4 (VNTR) of eNOS gene in malaria infected individuals of the population of Orissa (an eastern Indian state). PMID- 20004256 TI - Spatiotemporal dynamics of bilingual word processing. AB - Studies with monolingual adults have identified successive stages occurring in different brain regions for processing single written words. We combined magnetoencephalography and magnetic resonance imaging to compare these stages between the first (L1) and second (L2) languages in bilingual adults. L1 words in a size judgment task evoked a typical left-lateralized sequence of activity first in ventral occipitotemporal cortex (VOT: previously associated with visual word form encoding) and then ventral frontotemporal regions (associated with lexico semantic processing). Compared to L1, words in L2 activated right VOT more strongly from approximately 135 ms; this activation was attenuated when words became highly familiar with repetition. At approximately 400 ms, L2 responses were generally later than L1, more bilateral, and included the same lateral occipitotemporal areas as were activated by pictures. We propose that acquiring a language involves the recruitment of right hemisphere and posterior visual areas that are not necessary once fluency is achieved. PMID- 20004258 TI - Bioresorbable elastomeric vascular tissue engineering scaffolds via melt spinning and electrospinning. AB - Current surgical therapy for diseased vessels less than 6mm in diameter involves bypass grafting with autologous arteries or veins. Although this surgical practice is common, it has significant limitations and complications, such as occlusion, intimal hyperplasia and compliance mismatch. As a result, cardiovascular biomaterials research has been motivated to develop tissue engineered blood vessel substitutes. In this study, vascular tissue engineering scaffolds were fabricated using two different approaches, namely melt spinning and electrospinning. Small diameter tubes were fabricated from an elastomeric bioresorbable 50:50 poly(l-lactide-co-epsilon-caprolactone) copolymer having dimensions of 5mm in diameter and porosity of over 75%. Scaffolds electrospun from two different solvents, acetone and 1,1,1,3,3,3-hexafluoro-2-propanol were compared in terms of their morphology, mechanical properties and cell viability. Overall, the mechanical properties of the prototype tubes exceeded the transverse tensile values of natural arteries of similar caliber. In addition to spinning the polymer separately into melt-spun and electrospun constructs, the approach in this study has successfully demonstrated that these two techniques can be combined to produce double-layered tubular scaffolds containing both melt-spun macrofibers (<200microm in diameter) and electrospun submicron fibers (>400nm in diameter). Since the vascular wall has a complex multilayered architecture and unique mechanical properties, there remain several significant challenges before a successful tissue-engineered artery is achieved. PMID- 20004259 TI - Influence of moisture on the mechanical behavior of a natural composite. AB - The effects of moisture on the mechanical properties of the spicules of the sponge Euplectella aspergillum have been investigated. Determinations were made with the aid of a dynamic mechanical analyzer in both the static and dynamic modes, as well as imaging of the failed surfaces with scanning electron microscopy. For comparison purposes, melt-grown glass fibers of similar diameters were also studied in both distilled water and seawater. That exposure reduced both the stiffness and strength of the spicules. In addition, the energy required to achieve complete failure decreased in moist environments. The data for the wet spicules in both aqueous media showed decreasing values of energy dissipated until catastrophic failure compared to dry samples. The strength of wet glass decreased when compared with the dry condition, and the elastic modulus was also reduced. The most marked influence of moisture was seen in the damping effects in moist spicule samples that were nearly an order of magnitude larger than the damping of dry spicules. This effect was attributed mainly to plasticization of the thin organic layers. PMID- 20004260 TI - Engineering surfaces for site-specific vascular differentiation of mouse embryonic stem cells. AB - Differentiation of stem and progenitor cells routinely relies on the application of soluble growth factors, an approach that enables temporal control of cell fate but enables no spatial control of the differentiation process. Angiogenic progenitor cells derived from mouse embryonic stem cells (ESCs) were differentiated here according to the pattern of immobilized vascular endothelial growth factor-A (VEGF). Mouse ESCs engineered to express green fluorescent protein (eGFP) under control of promoter for the receptor tyrosine kinase Flk1 were used. The Flk1+ angiogenic progenitors were selected from day 3 differentiating embryoid bodies based on their expression of eGFP using fluorescence activated cell sorting. Mouse VEGF(165) was covalently immobilized onto collagen IV (ColIV) using 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide (EDC) chemistry. A non-cell adhesive layer of photocrosslinkable chitosan was first created, after which VEGF-ColIV was stamped as 100mum wide lanes on top of the chitosan layer and the Flk1+ angiogenic progenitors were seeded for site specific differentiation. Lanes stamped with only ColIV served as controls. The results presented here demonstrate that the cultivation of Flk1+ progenitors on surfaces with immobilized VEGF yielded primarily endothelial cells (53+/-13% CD31 positive and 17+/-2% smooth muscle actin positive), whereas surfaces without VEGF favored vascular smooth muscle-like cell differentiation (26+/-17% CD31 positive and 38+/-9% smooth muscle actin positive). PMID- 20004261 TI - A one-step method to fabricate PLLA scaffolds with deposition of bioactive hydroxyapatite and collagen using ice-based microporogens. AB - Porous poly(l-lactic acid) (PLLA) scaffolds with bioactive coatings were prepared by a novel one-step method. In this process, ice-based microporogens containing bioactive molecules, such as hydroxyapatite (HA) and collagen, served as both porogens to form the porous structure and vehicles to transfer the bioactive molecules to the inside of PLLA scaffolds in a single step. Based on scanning electron microscopy, energy-dispersive X-ray spectroscopy, X-ray diffraction and Fourier transform infrared spectroscopy analysis, the bioactive components were found to be transferred successfully from the porogens to PLLA scaffolds evenly. Osteoblast cells were used to evaluate the cellular behaviors of the composite scaffolds. After culturing for 8days, MTT assay and alkaline phosphatase activity results suggested that HA/collagen could improve the interactions between osteoblast cells and the polymeric scaffold. PMID- 20004263 TI - A pilot study on the use of natural calcium isotope (44Ca/40Ca) fractionation in urine as a proxy for the human body calcium balance. AB - We explored the possibility of using natural calcium (Ca) isotope variations in the urine (delta(44/40)Ca(urine)) as a proxy for the Ca balance in the human body. We chose two test persons extremely different in their health status, gender and age (4-year-old healthy boy and a 60-year-old woman known to suffer from osteoporosis). During a 5 day interval the Ca isotope composition of the individual diet (delta(44/40)Ca(diet)) was monitored for both test persons to be in general agreement to the Ca isotope composition of the normal western European diet ( approximately -1.02+/-0.1 per thousand). However, measurements showed that (1) delta(44/40)Ca(urine) of both test persons are approximately 1.37 and approximately 2.49 per thousand, respectively, heavier than delta(44/40)Ca(diet) and that (2) the delta(44/40)Ca(urine-boy) is approximately 1.1 per thousand heavier when compared to the value of the woman. The individual offset between diet and test persons is interpreted to reflect individual Ca reabsorption rates in the kidneys being the result of Rayleigh type Ca isotope fractionation related to the partitioning of Ca between the glomerular filtrate and filtered residue. The relative difference between delta(44/40)Ca(urine-boy) and delta(44/40)Ca(urine-woman) of approximately 1.1 per thousand may reflect individual differences in the balance of bone mineralization and demineralization processes related to age, gender and health status. By arbitrarily defining an equilibrium value for Delta(44/40)Ca(diet-urine) of -1.93 per thousand being the arithmetic mean of delta(44/40)Ca(urine) for both test persons the measured delta(44/40)Ca(urine) values may be applied to model the individual bone mineralization and demineralization processes in a qualitative way. Note, second order influences of intestinal Ca absorption during sequestration of Ca between intestine and blood have to be subject of further studies. PMID- 20004264 TI - Is kyphoplasty better than vertebroplasty in restoring normal mechanical function to an injured spine? AB - INTRODUCTION: Kyphoplasty is gaining in popularity as a treatment for painful osteoporotic vertebral body fracture. It has the potential to restore vertebral shape and reduce spinal deformity, but the actual clinical and mechanical benefits of kyphoplasty remain unclear. In a cadaveric study, we compare the ability of vertebroplasty and kyphoplasty to restore spine mechanical function, and vertebral body shape, following vertebral fracture. METHODS: Fifteen pairs of thoracolumbar "motion segments" (two vertebrae with the intervening disc and ligaments) were obtained from cadavers aged 42-96 years. All specimens were compressed to induce vertebral body fracture. Then one of each pair underwent vertebroplasty and the other kyphoplasty, using 7 ml of polymethylmethacrylate cement. Augmented specimens were compressed for 2 hours to allow consolidation. At each stage of the experiment, motion segment stiffness was measured in bending and compression, and the distribution of loading on the vertebrae was determined by pulling a miniature pressure transducer through the intervertebral disc. Disc pressure measurements were performed in flexed and extended postures with a compressive load of 1.0-1.5 kN. They revealed the intradiscal pressure (IDP) which acts on the central vertebral body, and they enabled compressive load bearing by the neural arch (F(N)) to be calculated. Changes in vertebral height and wedge angle were assessed from radiographs. The volume of leaked cement was determined by water displacement. Volumetric bone mineral density (BMD) of each vertebral body was calculated using DXA and water displacement. RESULTS: Vertebral fracture reduced motion segment compressive stiffness by 55%, and bending stiffness by 39%. IDP fell by 61-88%, depending on posture. F(N) increased from 15% to 36% in flexion and from 30% to 58% in extension (P<0.001). Fracture reduced vertebral height by an average 0.94 mm and increased vertebral wedging by 0.95 degrees (P<0.001). Vertebroplasty and kyphoplasty were equally effective in partially restoring all aspects of mechanical function (including stiffness, IDP, and F(N)), but vertebral wedging was reduced only by kyphoplasty (P<0.05). Changes in mechanical function and vertebral wedging were largely maintained after consolidation, but height restoration was not. Cement leakage was similar for both treatments. CONCLUSIONS: Vertebroplasty and kyphoplasty were equally effective at restoring mechanical function to an injured spine. Only kyphoplasty was able to reverse minor vertebral wedging. PMID- 20004262 TI - Manganese regulation of virulence factors and oxidative stress resistance in Neisseria gonorrhoeae. AB - Neisseria gonorrhoeae has evolved a complex and novel network of oxidative stress responses, including defence mechanisms that are dependent on manganese (Mn). We performed systematic analyses at the transcriptomic and proteomic (1D SDS-PAGE and Isotope-Coded Affinity Tag [ICAT]) levels to investigate the global expression changes that take place in a high Mn environment, which results in a Mn-dependent oxidative stress resistance phenotype. These studies revealed that there were proteins regulated at the post-transcriptional level under conditions of increased Mn concentration, including proteins involved in virulence (e.g., pilin, a key adhesin), oxidative stress defence (e.g., superoxide dismutase), cellular metabolism, protein synthesis, RNA processing and cell division. Mn regulation of inorganic pyrophosphatase (Ppa) indicated the potential involvement of phosphate metabolism in the Mn-dependent oxidative stress defence. A detailed analysis of the role of Ppa and polyphosphate kinase (Ppk) in the gonococcal oxidative stress response revealed that ppk and ppa mutant strains showed increased resistance to oxidative stress. Investigation of these mutants grown with high Mn suggests that phosphate and pyrophosphate are involved in Mn dependent oxidative stress resistance. PMID- 20004266 TI - Mifepristone: ten years later. AB - While pregnant women have sought abortifacients for thousands of years, they had no success at finding one that both worked and did not jeopardize their lives in the process. The discovery of mifepristone, with both anti-glucocorticoid and anti-progesterone properties, has had a profound effect on women's lives while weaving the abortion-related political hazards. Despite the controversies, millions of women around the world have used mifepristone for medical abortion. This review describes how researchers addressed the numerous barriers of a mifepristone abortion (i.e., gestational age limitation, lengthy process, high costs, complex regimen, failures, side effects and complications) and continue to improve upon the limited numbers and types of clinicians offering mifepristone. PMID- 20004265 TI - Immunization with the attenuated plasmidless Chlamydia trachomatis L2(25667R) strain provides partial protection in a murine model of female genitourinary tract infection. AB - Here we report on the safety, immunogenicity, and vaccine efficacy of the naturally occurring plasmid-free attenuated Chlamydia trachomatis L2-25667R (L2R) strain in a murine infection model. Intravaginal immunization induced both chlamydial specific serum antibody and systemic CD4(+) Th1 biased immune responses but failed to induce local IgA antibodies. Immunization induced no pathological changes in the urogenital tract. Protective immunity was evaluated by vaginal challenge with a natural occurring non-attenuated plasmid positive C. trachomatis urogenital strain (serovar D). Vaccinated mice were not protected from colonization/infection but exhibited a reduction in infectious burden at early time periods (1-2 weeks) post-challenge. Partial protective immunity did not protect against inflammatory disease. Thus, intravaginal vaccination with the live-attenuated L2R stain is safe, induces a systemic antibody and CD4(+) Th1 biased immune response, but its protective efficacy is limited to reducing chlamydial burden at early time periods post-infection. PMID- 20004267 TI - Review of clinical experience with estradiol in combined oral contraceptives. AB - Previous attempts to replace ethinylestradiol (EE) with 17beta-estradiol (E2) in combined oral contraceptives (COCs) have proved unsatisfactory in terms of bleeding outcomes. A review of previous studies of E2-based COCs has shown that, despite good ovulation inhibition, bleeding irregularities affected up to 100% of women, often resulting in high rates of discontinuation (up to 42%). Suggested reasons for the bleeding irregularities observed with these predominantly monophasic estradiol-progestin preparations included suboptimal doses of E2 and an inappropriate estrogen/progestin ratio. The progestin used in the investigated formulations (e.g., norethisterone acetate, desogestrel and cyproterone acetate) may also have affected the overall bleeding profile. More recent studies of a multiphasic COC containing estradiol valerate (E2V) and dienogest (DNG) indicate efficient ovulation inhibition and acceptable cycle control. In a randomized, double-blind trial that compared E2V/DNG with a monophasic COC comprising EE/levonorgestrel (LNG), the occurrence of scheduled withdrawal bleeding per cycle with E2V/DNG and EE/LNG was 77.7-83.2% and 89.5-93.8%, respectively. The intensity and duration of withdrawal bleeding was reduced with E2V/DNG. The incidence of intracyclic bleeding was similar with E2V/DNG (10.5-18.6%) and EE/LNG (9.9-17.1%). This review shows that after several unsatisfactory attempts to develop E2-based COCs, more recent studies employing endometrial-focused progestins, e.g., DNG, and multiphasic dosing regimens appear to be a promising approach for an E2-based COC that provides efficient ovulation inhibition and acceptable cycle control. PMID- 20004268 TI - Postmarketing study of ORTHO EVRA and levonorgestrel oral contraceptives containing hormonal contraceptives with 30 mcg of ethinyl estradiol in relation to nonfatal venous thromboembolism. AB - BACKGROUND: Concern has been raised that the risk of venous thromboembolism (VTE) in users of the ORTHO EVRA patch is higher compared to users of oral contraceptives (OCs). STUDY DESIGN: We identified idiopathic cases of VTE and controls, matched on age and index date, from among women in the United States PharMetrics/IMS and MarketScan databases who were current users of the patch or levonorgestrel-containing OCs with 30 mcg of ethinyl estradiol. We calculated odds ratios (ORs) and 95% confidence intervals (CIs). RESULTS: The ORs (95% CI) for VTE in users of the patch compared to levonorgestrel-containing OCs were 2.0 (0.9-4.1) and 1.3 (0.8-2.1) in the PharMetrics and MarketScan databases, respectively. ORs (95% CI) restricted to women aged 39 years or younger were 1.4 (0.6-3.0) and 1.2 (0.7-2.0), respectively. CONCLUSION: These results provide evidence that the risk of idiopathic VTE in users of the patch is not materially different than that of users of levonorgestrel-containing OCs in women aged 39 years or younger. We cannot rule out some increase in the risk in women aged 40 years or older. PMID- 20004269 TI - Oral contraceptive discontinuation and its aftermath in 19 developing countries. AB - BACKGROUND: The purpose of the article was to document oral contraceptive (OC) discontinuation and switching in a large number of low- and middle-income countries, and to assess the effects of women's education and reason for use (spacing vs. limitation). An attempt was made to explain intercountry variations. STUDY DESIGN: Calendar data from 19 Demographic and Health Surveys conducted between 1999 and 2005 were used. Data were analyzed by single- and multiple decrement life tables and by Cox proportional hazard model. RESULTS: The probability of stopping OC use within 12 months for reasons that implied dissatisfaction with the method ranged from 15% in Indonesia to over 40% in Bolivia and Peru with a median value of 28%. On average, 35% switched to a modern method within 3 months and 16% to a less effective 'traditional' method. Both education and reason for use were strongly related to the probability of switching to a modern method. Discontinuation was lower and switching higher in countries judged to have strong family planning programs. CONCLUSIONS: Both discontinuation of use and inadequate switching to alternative methods are major but neglected problems in the family planning services of many developing countries. PMID- 20004270 TI - Prospective study of weight change in new adolescent users of DMPA, NET-EN, COCs, nonusers and discontinuers of hormonal contraception. AB - BACKGROUND: Weight gain is commonly reported as a side effect of hormonal contraception and can lead to method discontinuation or reluctance to initiate the method. The purpose of this study was to investigate weight change in adolescent (aged 15-19 years) users of depot-medroxyprogesterone acetate (DMPA), norethisterone enanthate (NET-EN), combined oral contraceptives (COCs) and discontinuers of these methods as compared to nonusers of hormonal contraception. STUDY DESIGN: This longitudinal study recruited initiators of DMPA (n=115), NET EN (n=115), COCs (n=116) and nonusers of contraception (n=144). Participants were followed up for 4-5 years, and details of current contraceptive method, including switching, discontinuing and/or starting hormonal methods were documented at each 6-monthly visit. Women were classified according to their contraceptive histories on completion of the study, and injectable users were combined into one group for analysis. Height, weight and self-reported dieting were recorded at each visit. RESULTS: There was no difference in mean age or weight between the groups at baseline. Women using DMPA or NET-EN throughout, or switching between the two, had gained an average of 6.2 kg compared to average increases of 2.3 kg in the COC group, 2.8 kg in nonusers and 2.8 kg among discontinued users of any method (p=.02). There was no evidence of a difference in weight gain between women classified as nonobese or classified as overweight/obese in any of the four study groups at baseline. CONCLUSION: There is fairly strong evidence that adolescent contraceptive hormonal injectable users appear to gain more weight than COC users, discontinuers and nonusers of contraception. PMID- 20004271 TI - Oral contraceptive use and bone density in adolescent and young adult women. AB - BACKGROUND: Most of the millions of oral contraceptive (OC) users are under 30 years of age and in the critical period for bone mass accrual. STUDY DESIGN: This cross-sectional study of 606 women aged 14-30 years examined both OC duration and estrogen dose and their association with bone mineral density (BMD) at the hip, spine, and whole body (dual-energy X-ray absorptiometry). RESULTS: Of 389 OC users and 217 nonusers enrolled, 50% were adolescents (14-18 years). Of OC users, 38% used "low-dose" OCs [<30 mcg ethinyl estradiol (EE)]. In adolescents, mean BMD differed by neither OC duration nor EE dose. However, 19- to 30-year-old women's mean BMD was lower with longer OC use for spine and whole body (p=.004 and p=.02, respectively) and lowest for >12 months of low-dose OCs for the hip, spine and whole body (p=.02, .003 and .002, respectively). CONCLUSIONS: Prolonged use of today's OCs, particularly <30 mcg EE, may adversely impact young adult women's bone density while using these agents. PMID- 20004272 TI - The efficacy and safety of a low-dose, 91-day, extended-regimen oral contraceptive with continuous ethinyl estradiol. AB - BACKGROUND: This clinical trial was conducted to demonstrate the efficacy and safety of a 91-day extended-regimen, low-dose combination oral contraceptive (OC) consisting of 84 days of ethinyl estradiol (EE) 20 mcg/levonorgestrel (LNG) 100 mcg, followed by 7 days of 10 mcg EE in place of placebo. STUDY DESIGN: A multicenter open-label, single-treatment, Phase 3 study evaluated women aged 18 through 40 years over a treatment period of up to 1 year (four 91-day extended cycles). All subjects completed daily paper diaries to monitor compliance, bleeding and additional forms of contraception used during the course of the study. RESULTS: A total of 1249 subjects completed the study. The Pearl Index was 2.74 (95% confidence interval, 1.92-3.78), based on 36 pregnancies that occurred after the onset of treatment and within 14 days after the last combination tablet in women aged 18-35 years. Among compliant-use subjects 18-35 years old, the Pearl Index was 1.73 based on 22 on-treatment pregnancies. The life table pregnancy rate for subjects 18-35 years of age was 2.39%. Cycle control and adverse events reported with this regimen were similar to those reported with other low-dose OCs. CONCLUSIONS: This study demonstrated effective prevention of pregnancy with a 20-mcg EE, 91-day extended-regimen OC. In addition, the regimen was well tolerated and incidence of adverse events were consistent with what has been reported with other low-dose OCs. PMID- 20004273 TI - Efficacy of the low-dose combined oral contraceptive chlormadinone acetate/ethinylestradiol: physical and emotional benefits. AB - BACKGROUND: This study investigated the effects of the low-dose combined oral contraceptive (COC) 2.0 mg chlormadinone acetate (CMA)/0.03 mg ethinylestradiol (EE) (Belara, Balanca) on cycle-related physical and emotional disorders in women >or=25 years of age. STUDY DESIGN: A prospective, non-interventional, observational study of 3772 women over six cycles was conducted in 303 office based gynecological centers throughout Germany. RESULTS: CMA/EE provided high contraceptive efficacy with a Pearl index of 0 (95% confidence interval=0.00 0.22) and was generally well tolerated, with no statistically significant weight changes during the observation period (p=.147). CMA/EE intake resulted in a statistically significant improvement in cycle-related physical and emotional symptoms, with a 67% overall reduction in sum score for number and intensity of cycle-related symptoms per patient. CONCLUSIONS: The results of this study in women >or=25 years of age support previous findings that 2.0 mg CMA/0.03 mg EE is an effective low-dose COC, with an excellent tolerability profile, with the additional benefits of significantly reducing both cycle-related physical and emotional symptoms (por=500 IU/L than in women whose serum beta-hCG was <500 IU/L (p<.001), and the overall sensitivity and specificity of serum beta-hCG (>or=500 IU/L) were 97.1% and 62.5%, respectively. Moreover, the incomplete abortion rate was greater in women with an endometrial thickness >or=15 mm than in women with an endometrial thickness <15 mm (p<.001), and the overall sensitivity and specificity of endometrial thickness (>or=15 mm) were 94.1% and 75%, respectively. No complication occurred. CONCLUSIONS: The combination of mifepristone and misoprostol was found to be a safe and effective method to terminate early pregnancy in women with a previous cesarean delivery. If a woman with a prior uterine incision experienced vaginal bleeding intervals >or=21 days and/or had a bilayer endometrial thickness >or=15 mm and/or serum beta-hCG >or=500 IU/L after a medical abortion, then she should undergo curettage. PMID- 20004276 TI - A randomized pilot study on the effectiveness and side-effect profiles of two doses of digoxin as fetocide when administered intraamniotically or intrafetally prior to second-trimester surgical abortion. AB - BACKGROUND: Digoxin is commonly used to facilitate second-trimester surgical abortion despite limited data regarding its safety and effectiveness for this indication. We conducted a pilot study to determine the incidence of side effects after digoxin administration and whether effectiveness can be improved with variations in dose and technique. STUDY DESIGN: Fifty-two women presenting for elective termination of pregnancy between 18 and 24 weeks' gestation were randomized to one of four digoxin treatment groups: 1.0 mg intraamniotic (1.0 IA), 1.0 mg intrafetal (1.0 IF), 1.5 mg intraamniotic (1.5 IA) or 1.5 mg intrafetal (1.5 IF). Ultrasound was used to assess for the presence of fetal cardiac activity prior to the abortion procedure. Data on the presence and severity of pain, nausea and other potential side effects were collected before digoxin injection, immediately following digoxin injection and on the day after digoxin injection. RESULTS: Digoxin effectively induced fetal death in 87% of women. The failure rate did not vary by route of administration (IA or IF) and was not lowered by increasing the dose from 1.0 to 1.5 mg. IF injections induced fetal death more rapidly than IA injections. Digoxin administration did not result in increased pain or nausea. CONCLUSIONS: IA or IF injection of digoxin is safe and effective for inducing fetal death prior to second-trimester surgical abortion. Doses greater than 1.0 mg may not be necessary. PMID- 20004277 TI - Cancer risk after sterilization with transcervical quinacrine: updated findings from a Chilean cohort. AB - BACKGROUND: Dating back to the 1970s, thousands of women worldwide have voluntarily been sterilized with transcervical insertion of quinacrine pellets. The safety and efficacy of the technology are still being assessed today; in particular, better estimates on the incidence of human cancers are now feasible. METHODS: We conducted a cohort study of 1492 women in Santiago and Valdivia, Chile, who received transcervical quinacrine pellets for contraceptive sterilization between 1977 and 1989. We periodically interviewed women with the last interviews in 2006-2007 and reviewed their medical records. We calculated age and site-specific incidence of invasive cancers and compared the observed cases to the number of expected cases based on data from the Cali, Colombia, cancer registry, gathered by the International Agency for Research on Cancer. RESULTS: During 23,894 person-years of follow-up, 41 invasive cancers were identified, including 16 new cases that had occurred since the previous analysis. Ten cases of cervical cancer were observed, compared with 12.1 expected. Since the initial study's confirmation of a single case of leiomyosarcoma, no other uterine cancers have been diagnosed. We would expect 2.0 uterine cancers during this number of observed women-years. One case of ovarian cancer was diagnosed, compared with 3.1 expected. CONCLUSION: Rates of cancer among women exposed to intrauterine quinacrine are similar to population-based rates. PMID- 20004278 TI - A randomized, double-blind, placebo-controlled Phase II extended safety study of two Invisible Condom formulations in Cameroonian women. AB - BACKGROUND: Invisible Condom gel formulations being developed as microbicides to prevent the sexual transmission of HIV are advancing through the phases of clinical trials. The objectives of this study were to evaluate, after 8 weeks of vaginal application, the extended safety and acceptability of two Invisible Condom vaginal gel formulations: (i) the polymer alone and (ii) the polymer containing sodium lauryl sulfate (SLS) compared to placebo. STUDY DESIGN: This study is a randomized, doubled-blind, placebo-controlled Phase II extended safety study in healthy sexually active women from Yaounde, Cameroon. Women were randomized into three gel arms: (i) placebo, (ii) polymer alone and (iii) polymer/SLS. Women applied gel intravaginally twice daily for 8 weeks. RESULTS: A total of 194 sexually active women applied placebo (n=41), polymer alone (n=76) and polymer/SLS (n=77). Invisible Condom gel formulations were well tolerated with no reported serious adverse events. The majority of reported adverse events were mild or moderate and mostly similar in all three arms, except for pelvic pain that was 10% higher in the polymer and polymer/SLS arms compared to placebo. Colposcopy showed neither genital ulceration nor mucosal lesions. Nugent score, H(2)O(2)-producing lactobacilli and vaginal pH were not affected by the study products. The gel formulations and applicator were generally acceptable and comfortable. CONCLUSION: This extended safety study showed that the Invisible Condom gel formulations and applicator were well tolerated and acceptable when applied intravaginally twice daily for 8 weeks. Thus, further phases of clinical development of Invisible Condom as a potential microbicide to prevent sexual transmission of HIV are warranted. PMID- 20004279 TI - Loop electrosurgical excision procedure to treat cervical stenosis at the time of surgical abortion. AB - Cervical stenosis can act as a barrier to the uterine cavity for surgical abortion. We present a case in which a loop electrosurgical excision procedure (LEEP) was performed during a surgical abortion to overcome a stenotic cervix. We also discuss risk factors for cervical stenosis, and other methods of treating stenosis which have been reported in the literature. PMID- 20004280 TI - Effects of the contraceptive vaginal ring, the contraceptive transdermal patch and combined oral contraceptives on markers of hemostasis. PMID- 20004282 TI - [Risk of homicide and major mental disorders: a critical review]. AB - INTRODUCTION: Tragic and high profile killings by people with mental illness have been used to suggest that the community care model for mental health services has failed. It is also generally thought that schizophrenia predisposes subjects to homicidal behaviour. OBJECTIVE: The aim of the present paper was to estimate the rate of mental disorder in people convicted of homicide and to examine the relationship between definitions. We investigated the links between homicide and major mental disorders. METHODS: This paper reviews studies on the epidemiology of homicide committed by mentally disordered people, taken from recent international academic literature. The studies included were identified as part of a wider systematic review of the epidemiology of offending combined with mental disorder. The main databases searched were Medline. A comprehensive search was made for studies published since 1990. RESULTS: There is an association of homicide with mental disorder, most particularly with certain manifestations of schizophrenia, antisocial personality disorder and drug or alcohol abuse. However, it is not clear why some patients behave violently and others do not. Studies of people convicted of homicide have used different definitions of mental disorder. According to the definition of Hodgins, only 15% of murderers have a major mental disorder (schizophrenia, paranoia, melancholia). Mental disorder increases the risk of homicidal violence by two-fold in men and six-fold in women. Schizophrenia increases the risk of violence by six to 10-fold in men and eight to 10-fold in women. Schizophrenia without alcoholism increased the odds ratio more than seven-fold; schizophrenia with coexisting alcoholism more than 17 fold in men. We wish to emphasize that all patients with schizophrenia should not be considered to be violent, although there are minor subgroups of schizophrenic patients in whom the risk of violence may be remarkably high. According to studies, we estimated that this increase in risk could be associated with a paranoid form of schizophrenia and coexisting substance abuse. The prevalence of schizophrenia in the homicide offenders is around 6%. Despite this, the prevalence of personality disorder or of alcohol abuse/dependence is higher: 10% to 38% respectively. The disorders with the most substantially higher odds ratios were alcohol abuse/dependence and antisocial personality disorder. Antisocial personality disorder increases the risk over 10-fold in men and over 50-fold in women. Affective disorders, anxiety disorders, dysthymia and mental retardation do not elevate the risk. Hence, according to the DMS-IV, 30 to 70% of murderers have a mental disorder of grade I or a personality disorder of grade II. However, many studies have suffered from methodological weaknesses notably since obtaining comprehensive study groups of homicide offenders has been difficult. CONCLUSIONS: There is an association of homicide with mental disorder, particularly with certain manifestations of schizophrenia, antisocial personality disorder and drug or alcohol abuse. Most perpetrators with a history of mental disorder were not acutely ill or under mental healthcare at the time of the offence. Homicidal behaviour in a country with a relatively low crime rate appears to be statistically associated with some specific mental disorders, classified according to the DSM-IV-TR classifications. PMID- 20004283 TI - [Social phobia in anorexia nervosa: evolution during the care]. AB - CONTEXT: The links between anorexia nervosa (AN) and anxiety disorders, and particularly social phobia, are little known. However, social phobia occurs frequently in AN. Some studies have shown reduction in anxious and depressive symptomatology in AN with re-nutrition. But, to our knowledge, no work has examined the evolution of social phobia symptoms during re-nutrition in AN. OBJECTIVES: To specify the links between AN, nutritional state, and social phobia. METHOD: The population consisted of 2 samples and the analysis was conducted using the SPSS11.5. Sample 1 (N=24 AN) was evaluated on admission and on leaving the hospital. Our evaluation used the body mass index (BMI), the Liebowitz scale, the Mini International Neuropsychiatric Interview (MINI), and the Yale-Brown Obsessive Compulsive Scale for Eating Disorders scale (Y-BOCS-ED) respectively to evaluate or diagnose the state of malnutrition, social anxiety symptomatology, social phobia in Diagnostic and Statistical Manual-4 (DSM-IV) and anorexic symptomatology. Sample 2 (N=60) was assessed at the end of the hospitalization and then 6, 12 and 18 months later. We used the BMI, Liebowitz scale, MINI, and Eating Disorders Inventory (EDI) to assess anorexic symptomatology. In addition, the Morgan-Russell outcome assessment schedule (MR schedule) was used to assess the total clinical state of the patients. RESULTS: Social anxiety symptomatology and actual diagnosis decreased throughout the treatment. However, regardless of the point at which the patient received care, there was no correlation between social phobia and nutritional state, as indicated by BMI. A correlation existed between social phobia and AN symptomatology, and between social phobia and total clinical state, during the out-patient care. CONCLUSION: A component of AN-social phobia comorbidity is still questionable. Is it linked to the clinical state of the subjects (question of an additional effect of under nutrition and cognition), or even to AN? Others indicators of under nutrition are of interest and warrant further evaluations. We therefore feel that a diagnosis of social phobia can only be confirmed after an acute state of AN, thus allowing for preferential treatments. Others studies must be conducted in order to continue to explore the links between social phobia and AN. PMID- 20004284 TI - [Self-mutilating behaviour: a study on 30 inpatients]. AB - INTRODUCTION: Deliberate self-injury is defined as the intentional, direct injuring of body tissue without suicidal intent. There are different types of deliberate self-mutilating behaviour: self cutting, phlebotomy, bites, burns, or ulcerations. Sometimes, especially among psychotic inpatients, eye, tongue, ear or genital self-mutilations have been reported. In fact, self-mutilation behaviour raises nosological and psychopathological questions. A consensus on a precise definition is still pending. Many authors consider self-mutilating behaviour as a distinct clinical syndrome, whereas others hold it to be a specific symptom of borderline personality disorder. Self-mutilating behaviour has been observed in 10 to 15% of healthy children, especially between the age of 9 and 18 months. These self mutilations are considered as pathological after the age of 3. Such behaviour is common among adolescents, with a higher proportion of females, and among psychiatric inpatients. Patients use different locations and methods for self-mutilation. Deliberate self harm syndrome is often associated with addictive behaviour, suicide attempt, and personality disorder. CLINICAL MATERIAL: We report on an observational study including 30 inpatients and we compared the data with the existing literature. As a matter of fact, until now, most of the papers deal with case reports or with very specific patterns of self mutilation (eye, tongue or genital self-mutilations). Otherwise, papers report the relationships between self-mutilation and somatic or personality disorders (Lesh Nyhan syndrome, borderline personality disorder, dermatitis artefacta, self mutilation in children following brachial plexus related to birth injury, mental retardation...). Our study included all self harmed patients who had been admitted to our psychiatric hospital (whatever the location and type of self mutilation). Patients suffering from brain injury or mental retardation were excluded. RESULTS: In our sample, there was a higher percentage of women (29 women and 1 man) and the mean age was 18 (12 to 37). More than half of the patients were aged under 18. Single parent families were reported in 30% of cases. Thirty percent of patients had been physically or sexually abused during childhood. Sixty percent had a comorbid psychiatric disorder, 63% had been hospitalised previously (half of them twice or more). Seventy-three percent of patients had previously attempted suicide (notably deliberate self-poisoning and cutting) that was not considered as self-mutilating behaviour by the patients themselves. Each patient had self harmed themselves at least twice and most often different methods and locations were used (deliberate self harm of forearms 90%, thighs 26.7%, legs 16.7%, chest 10%, belly 10%, hands 6.9%, face 6.9%, arms 6.7%, and feet 3.3%). Addictive disorders, such as substance abuse (tobacco 46.7%; alcohol 23.3%; illicit drugs 16.7% mostly cannabis or cocaine) and eating disorders (33.3% and among them 50% of cases were restrictive anorexia nervosa) were often associated with a deliberate self harm syndrome. Three psychiatric diagnoses were often observed in our cohort: depressive disorder 36.7%; personality disorder 20%; psychosis 10% and depressive disorder associated with personality disorder 33.3%. In our sample, psychotic patients differed on several clinical aspects: the atypical location (abdomen, nails) and method (needles) of self-mutilating behaviour. None of them had been abused during childhood and none was suffering from addictive disorders. PMID- 20004285 TI - [Preliminary comparative study of the personality disorder evaluation DIP instrument with the semi-structured SCID-II interview]. AB - INTRODUCTION: This work deals with the comparative study of two standardised instruments, which can be used to diagnose personality disorders (PD): the SCID II and the DIP. Each instrument used as a self-questionnaire followed by a semi structured interview by the same clinician was applied to 21 patients suffering from PD. The DIP (DSM-IV and ICD-10 Personality), which is a recent instrument, consists of a self-questionnaire (DIP-Q) and a semi-structured interview (DIP-I), created by Bodlund and Ottosson. It makes it possible to evaluate PD from criteria based on the DSM-IV as well as the ICD-10. We translated it into French then evaluated it in comparison with another instrument, the Structured Clinical Interview for DSM-IV Axis II PD (SCID-II) whose validity was demonstrated by Bouvard. METHOD: For the self-questionnaire (SCID-auto), we used CUNGI'S computerised version. The present version of the semi-structured interview SCID-E (French translation by Bouvard et al.) evaluates the 10 PD of the DSM-IV, the depressive personality and the passive-aggressive personality, included in the DSM-IV appendix B. The DIP-Q questionnaire is made up of 140 right/wrong items referring to the 10 PD of the DSM-IV and the eight disorders of the ICD-10. The DIP-I is the self-structured interview created by Ottosson et al. and it is built on the same pattern as the SCID-II. It provides diagnoses for all DSM-IV and/or ICD-10 PD as well as the schizotypic disorder. The DIP-I is usually preceded by a general "scan" interview in order to assess an existing personality disorder corresponding to Axis I of the DSM-IV or the ICD-10. In our study, we substituted a Mini International Neuropsychiatric Interview (MINI) questionnaire for this interview. Twenty-four patients suffering from one or several PD were chosen among ambulatory or out-patients by clinicians from the Saint-Etienne Psychiatric University Hospital Center. The diagnosis was not revealed to the examiner during the study. The subjects filled in the DIP-I and the SCID-II self-questionnaires. The answers to each test were first processed through a computer, then the patients were seen over the following weeks for the DIP-I and SCID-II semi structured interviews. For both questionnaires, we only explored the diagnostic categories reaching pathological level (as was recommended by the authors). Considering the small number of patients involved, we used nonparametric tests: Wilcoxon test, Mac Nemar test and the Kappa. RESULTS: As far as the self questionnaire results are concerned, we noticed important differences for the schizoid and the schizotypic PD between the DIP-Q (ICD) and the DIP-Q (DSM). The most represented PDs are the paranoiac, borderline, avoiding and obsessional personalities. After the semi-structured interviews, it appears that only 30 to 50% of the diagnoses obtained through self-evaluation were confirmed (with the exception of the schizotypic personality and the antisocial personality for the SCID with perfect agreement between self and clinical evaluation). Globally, the agreement between diagnosis by self-evaluation and diagnosis by semi-structured interview is not very satisfactory. Finally, a cluster analysis of the results of the three semi-structured interviews put together reveals that five patients show at least one PD diagnosed in the three clusters, two have no diagnosis, six patients have one or several PDs in clusters B and C, three patients have some in clusters A and C, and five patients only have some in cluster C. Our results lead to several remarks: the size of our group is small, but it must be pointed out that the investigations for each patient took about three hours, which made it difficult for the patients to agree when the clinicians proposed the study; three patients originally included could not be evaluated because of suicidal behaviour. In their self-administered form, the SCID and the DSM version of the DIP-Q broadly diagnose a little more than three PDs per patient, whereas the ICD version of the DIP-Q diagnoses more than five. The administration of semi structured interviews leads to an average of 1.3 diagnosis for the DIP-Q DSM-IV and 1.6 for the ICD against 1.9 PD for the SCID interview. These results correspond to the literature data. There are differences between the SCID and the DIP-I, as regards to the way they were used: the SCID-II makes it necessary to repeat the questions positively answered in the self-questionnaire, whereas the DIP-I explores all the criteria of the whole diagnosed PD, which may favour the inclusions. Concerning other instruments compared to the SCID-II in the international literature, our results with the DIP are globally satisfactory. CONCLUSION: The results must be interpreted with some care, considering the small number of patients. Important discrepancies were noticed between the diagnoses obtained through self-evaluation and the semi-structured interview, mainly for the A and C personality clusters of the DSM-IV, showing the tests to be extremely sensitive, but not specific enough for detection. However, the agreement between both instruments referring to the DSM-IV is satisfactory. The main interest of our work was to make the first French translation of the DIP known and to compare it to another instrument, which has often been evaluated previously. PMID- 20004286 TI - [Frequency and risk factors for pathological gambling in a sample of gamblers in Casablanca, Morocco]. AB - INTRODUCTION: Pathological gambling is a major psychiatric disorder and a public health problem that has gained a lot of attention in the last few years. The problems caused by gambling are increasingly serious. The prevalence of pathological or compulsive gambling varies from 0.8 to 2% in Europe, Canada and the USA. AIM OF THE STUDY: A study has been conducted in different gambling spots in Casablanca to measure the frequency of pathological gambling in Morocco and to determine the risk factors and the socio-cultural factors associated with it. DESIGN OF THE STUDY: The collection of data was conducted in the most frequented gambling spots. The first part of the questionnaire permitted the gathering of the sociodemographic characteristics and drug addictions as well as the psychiatric and legal histories. The detection and diagnosis of pathological gambling was conducted using through the South Oaks Gambling Screen (SOCS) questionnaire, which has been translated to Arabic to suit the needs of the study. PARTICIPANTS IN THE STUDY: Two hundred men were retained out of the 243 interviewed, which represent a participation rate of 82%. The frequency of pathological gambling among a population of gamblers is 53%. The mean age of the sample was 42.3 more or less 10.70 years. Thirty-six percent of those interviewed had a monthly income of less than 2000 dirhams (200 euros). Horse and greyhound racing were the most popular forms of gambling among those interviewed, with rates of 91 and 60% respectively. Individuals with an educational level of no more than primary school, a monthly income of less than 5000 dirhams (500 euros), a personal psychiatric history, and a drug addiction, are most risky to be pathological gamblers. CONCLUSION: The topic pathological gambling has been studied very little in Arab and Islamic countries, and it is necessary to conduct larger epidemiological studies on the general population to inquire about its prevalence, risk factors as well as its psychiatric characteristics. PMID- 20004287 TI - [Prevalence of psychiatric disorders in French general practice using the patient health questionnaire: comparison with GP case-recognition and psychotropic medication prescription]. AB - INTRODUCTION: Psychiatric disorders, mainly depression and anxiety, are frequently encountered in primary care and are a major cause of distress and disability. Nearly half of cases go unnoticed and among those that are recognised, many do not receive adequate treatment. In France, there is limited research concerning the prevalence, detection and management of these conditions in primary care. OBJECTIVES: To estimate the prevalence of psychiatric disorders, overall and for the main psychiatric diagnostic categories, encountered in primary care; to describe general practitioners' (GPs') case identification rate; to examine psychotropic medication prescription according to diagnosis, in a regionally representative sample of GP attenders. METHODS: GP practicing standard general practice in an urban area of the city of Montpellier and a nearby semi rural region were recruited to participate. The response rate was 32.8% (n=41). Five additional GP almost exclusively offering homeopathy and acupuncture were recruited nonrandomly for convenience purposes. In each GP surgery, consecutive patients entering the waiting room were invited by a research assistant to participate until 25 patients per GP were recruited. Each participant completed self-report questionnaires in the waiting time, including the patient health questionnaire (PHQ), which yields provisional DSM-IV diagnoses. The GP completed a brief questionnaire during the consultation, giving his/her rating of the severity of any psychiatric disorder present and action taken. RESULTS: The patient response rate was 89.8%. In all, 14.9% of patients reached DSM-IV criteria for major depression or anxiety disorder on the PHQ (9.1% for major depression, 7.5% for panic disorder; 6% for other anxiety disorders). For the subthreshold categories, 7.4% met criteria for other depressive disorders, 11.8% for somatoform disorders and 10.9% for probable alcohol abuse or dependence. 66.3% of patients with DSM-IV diagnoses of major depression or anxiety disorder were identified by the GP as having a psychiatric disorder. The identification rate was 51% for all depressive disorders, anxiety and somatoform disorders. Of patients receiving a prescription for anxiolytic or antidepressant medication on the survey day, 80% were classified as cases of psychiatric disorder by the GP. Only 48.8% met criteria for major depression or anxiety disorder on the PHQ. CONCLUSION: This study highlights the frequency of psychiatric disorders in a regional study of French general practice. Overall, prevalence rates were similar to those found elsewhere, except for probable alcohol abuse and dependence, which was considerably higher than in the USA PHQ validation study. As in other countries, GP identified roughly half of psychiatric cases. Furthermore, half of patients treated by anxiolytic or antidepressant medication did not meet the diagnostic criteria on the survey day for which these medications have mainly shown their efficacy. This confirms the French paradox of one of the highest psychotropic medication consumption rates in Europe despite many cases of depression remaining untreated. The PHQ could be a rapid and acceptable diagnostic aid tool for French general practice but first needs to be validated against the diagnosis of mental health professionals in this setting. PMID- 20004288 TI - [Screening for bipolar disorders: a review of the literature]. AB - OBJECTIVE: The aim of this article is to review the major instruments proposed for screening for bipolar disorder among clinical or general, adult or paediatric populations. They were developed in order to improve the detection of this illness which, far too often, remains unrecognized. Several of these screening instruments are already translated into several languages and validated. METHODOLOGY: A systematic review of the literature published on this topic up to July 2007 was carried out, using the main electronic data base (Medline). The keywords employed included bipolar disorder, screening, questionnaire, diagnosis and early recognition. RESULTS: The studies reported here examine whether screening instruments perform similarly in various clinical and non-clinical samples. Different forms of the same questionnaire (like self-report or parent report used in paediatric samples) are sometimes compared, usually showing that parent reports supersede the adolescent self-report form. This is namely the case for the Mood Disorder Questionnaire (MDQ) which is a brief and widely tested tool, available both in adult and adolescent versions. The MDQ exhibits good psychometric properties in relation to sensitivity and specificity in adult psychiatric samples, but these are more limited in the general population. Moreover, it yields better sensitivity for BP type I than for other bipolar subtypes. This is also true for other screening instruments like the hypomania check list (HCL-32). In order to optimize the sensitivity for bipolar II disorders, proposals for changing the MDQ screening algorithm have been tested. DISCUSSION: Even though it does not replace a thorough clinical interview, the use of screening tools for bipolar disorder is widely advocated. We discuss the need for clinicians to rely upon instruments allowing for a rapid and economically feasible identification of this disorder. Involving family members in the evaluation process may also increase the rate of recognition. More studies are still required in order to improve diagnostic efficiency of the screening instruments. PMID- 20004289 TI - [Efficacy of escitalopram and severity of depression: new data]. AB - According to recent data in the literature, severe forms of depression represent almost half of all characterised episodes of depression; in view of this their treatment is a real public health challenge. According to the recommendations of various health authorities, treatment first relies on antidepressant medication. A recent meta-analysis of 12 new generation antidepressants has shown that these products are not all equivalent in the treatment of major depression, and that differences exist in terms not only of efficacy but also of acceptability. These differences in efficacy also appear true in the treatment of severe depression. Very recent publications, with a meta-analysis carried out on large cohorts of patients included in clinical trials, have shown the superior efficacy of escitalopram (which is the most active enantiomer of the racemic component, citalopram) in severe depression (defined by a total score on the MADRS scale > or =30). A pooled analysis by Kilts et al. assessed the response to treatment according to the baseline severity of the depression, under escitalopram and under six comparable products: citalopram, duloxetine, fluoxetine, paroxetine, sertraline, and venlafaxine. It showed that the rate of responders to the different treatments decreased when baseline severity increased, which agrees with the data in the literature, except for escitalopram, with which the rate of responders remained stable. The pooled analysis of Kennedy et al., that partly referred to the same studies, showed that for all the patients exhibiting severe depression, escitalopram was significantly superior to the other same six comparable products (mean estimated difference of 1.8 on the total MADRS score [p<0.0001]; a rate of 64.4 responders vs 55.8% [odds ratio=1.60, p<0.0001]; rate of remission of 47.7 vs 41.6% [odds ratio=1.39, p<0.0007]). In parallel with the statistical significance of the results, the criterion of clinical pertinence of the differences revealed between the medicinal products is highly significant. Regarding antidepressants, the criteria of clinical pertinence most frequently applied are the difference in rate of responders, the number needed to treat (NNT) and, to a lesser extent, the difference between the rates of patients in remission and the difference in the effect of treatment. According to these criteria, Montgomery and Moller (2009) assessed the clinical pertinence of the results showing the superior efficacy of escitalopram on three products: citalopram, paroxetine and duloxetine. Hence, they established this clinical pertinence for all of the depressed patients, but also in the particular case of severe depression. All these new data confirm the interest of escitalopram in the treatment of characterised depression and notably in the severe forms, with a particularly favourable efficacy/acceptability ratio. PMID- 20004290 TI - [Persons suffering from schizophrenia and relapses]. AB - INTRODUCTION: In schizophrenia, relapse is a common event that affects more than half the patients within 2 years after a first episode. It is a real setback for them and their relatives. Surprisingly, we do not have much information on how patients and their relatives experience the relapse. METHOD: A national survey was conducted among 316 schizophrenic outpatients treated with antipsychotics, and 82 of their relatives. The survey assessed the following four aspects: disease history, last relapse history, hospitalization experiences, and relapse prevention. RESULTS: Regarding the disease history, the average psychiatric follow-up was 13 years and patients had been hospitalized five times on average. Relatives reported approximately the same history. Regarding the last relapse, 9/10 of relatives reported that this relapse led to hospitalization and 69% of patients understood that their hospitalizations were due to relapse. 4% of patients and 7% of relatives identified the end of the treatment as a precursor to relapse. While a lack of compliance was found in about four relapses out of 10. It has also been shown that patients confided primarily in the medical team and the relatives thought to be the first confidant of patients. Regarding the experience of hospitalization, 87% of patients and 86% of relatives judged the hospitalization useful. For both, hospitalization represented a solving step to manifestations of relapse. Regarding the relapse prevention, almost three patients out of four thought they knew what to do in order to avoid a new relapse, while only 52% of the relatives thought patients knew what to do for this matter. For more than one third of the patients, the last relapse (3 years ago) was still a painful event. Avoiding a new relapse was considered very important or important by 91% of patients and 100% of relatives. Relatives felt that regular appointments with the medical team helped avoid relapses. Fifty-nine per cent of relatives have said it was difficult to verify whether or not the treatment was taken by a schizophrenic patient. Relatives' opinion on the injectable treatment was favorable and approximately 50% of the patients declared knowing of injectable treatments. Among these 72% felt that such treatment was reassuring, 69% said it was simpler than oral therapy, and 67% thought it was the most suitable to check the compliance. Only 31% considered it restricting for the patient, against 54% who were considering it not restricting. Finally 57% of patients were willing to take an injectable treatment in order to prevent further hospitalization. CONCLUSION: This study brings us a better understanding of patients' and relatives' experience of relapse. These results demonstrate the potential impact of relapse on the patients and their relatives and highlight their motivation to avoid further relapses. Also revealed, the lack of importance given to the link between compliance and relapse by patients and relatives. These results underscore the complexity of this disease management in which each player has a key role. PMID- 20004291 TI - [Milnacipran and venlafaxine at flexible doses (up to 200 mg/d) in the outpatient treatment of adults with moderate-to-severe major depressive disorder: a 24-week randomised, double blind exploratory study]. AB - INTRODUCTION: Serotonin (HT) and noradrenaline (NA) reuptake inhibitors (SNRIs) are commonly used as first line treatment of major depressive disorders (MDD). As compared to tricyclic antidepressants, they have proved similar efficacy and better tolerability. Milnacipran (MLN) (Ixel) and venlafaxine (VLF) (Effexor) are two SNRIs pharmacologically differing by their NA/HT ratio of potency: 1:1 and 1:30, respectively. OBJECTIVES: To investigate the efficacy and safety/tolerability of MLN and VLF administered at flexible doses (100, 150 or 200 mg/day) for 24 weeks (including 4 weeks of up-titration) in the outpatient treatment of adults with moderate-to-severe MDD. DESIGN: Multicentre, randomised, double blind, 2-parallel-arm, 24-week exploratory trial conducted in France by 50 psychiatrists. DIAGNOSIS AND MAIN INCLUSION CRITERIA: Male or female outpatients, aged 18 to 70, meeting the DSM-IV-TR and related MINI criteria for recurrent, unipolar, moderate-to-severe MDD, with neither psychotic features nor severe suicidal risk. A Montgomery-Asberg depression rating scale (MADRS) score> or =23 was required at inclusion. TREATMENT SCHEDULE: Patients were randomised to receive either MLN or VLF (1:1 ratio) for 24 weeks in double-blind conditions. Regardless of the treatment received, the following dosing schedule was applied: during the initial 4-week up-titration phase, the dosage was progressively increased from 25 mg/day (qd administration) to 150 mg/day (bid administration). At week 4, the dosage was either maintained at 150 mg/day, or adapted to 100 or 200 mg/day, based on the investigator's clinical judgement. At any time during the 20 following treatment weeks, the dose could be lowered for safety concerns until a minimal threshold of 100 mg/day. From Week 24, the dosage was decreased by 50mg/day every five days. After randomisation, eight assessment visits were organised at 2, 4, 6, 8, 12, 18, 24 weeks, and at study end (after the 5-15 days of down-titration and 10 days free of treatment). Efficacy evaluation ratings included the MADRS and global disease severity (CGI-S) total scores. Rates of MADRS response (reduction of initial score> or =50%) and remission (score< or =10) were calculated at Week 8 and Week 24 in the full analysis set as well as in the subgroups of patients with depressive disorder of severe DSM-IV intensity and with a MINI evaluation of suicidal risk (rated as required 'moderate' at the worst). STATISTICAL ANALYSIS: Standard distribution statistics (including mean and standard deviation [S.D.]) of scores and their changes from baseline, were calculated using the observed-case (OC) approach at all assessment times for the MADRS score, and the last-observation-carried-forward (LOCF) at 8 and 24 weeks for both MADRS and CGI-S scores. MADRS response and remission rates at 8 and 24 weeks were calculated using the LOCF approach by normal approximation of the binomial distribution. Bilateral exploratory statistical tests at 5% significance level were performed for results at 8 and 24 weeks of: (i) MADRS score changes from baseline, based on the score progress at each visit (mixed model for repeated measurements [MMRM]), and (ii) global MADRS response and remission rates (Chi(2)). RESULTS AND PATIENTS: A total of 195 patients were randomly assigned MLN (n=97) or VLF (n=98) and 134 (68.7%: 61.9%/MLN and 75.5%/VLF) completed the trial. At the end of the up-titration, patients received 100 mg/day (11.4%/MLN, 10%/VLF), 150 mg/day (30.4%/MLN, 43.8%/VLF), or 200 mg/day (58.2%/MLN, 46.3%/VLF). Totals of 177 patients (90/MLN and 87/VLF) and 181 patients (90/MLN and 91/VLF) were analysed for efficacy and safety, respectively. Treatment groups were similar for baseline characteristics except a higher proportion of MLN patients with a severe depressive episode (63.3% versus 54%). RESULTS AND EFFICACY: MADRS score (mean [S.D.] initial score: 31 [4.5]) progressively decreased all along the treatment course and similarly in both groups (Week 8-OC : -18.8 [7.7]/MLN and -18.6 [7.3]/VLF, p(MMRM)=0.95 ; Week 24-OC : -23.1 [7.8]/MLN and -22.4 [7.3]/VLF, p(MMRM)=0.37 ). At week 8-LOCF, MADRS response rates were similar in both groups (64.4%/MLN, 65.5%/VLF, p(chi2)=0.88) as well as remission rates (42.2%/MLN, 42.5%/VLF p(chi2)=0.97). At week 24 they remained non clinically and statistically different between groups (response rates: 70%/MLN, 77%/VLF, p(chi2)=0.29; remission rates: 52.2%/MLN, 62.1%/VLF, p(chi2)=0.19). In both "severe depressive episode" and "MINI mild or moderate suicidal risk" subgroups (n=104 and 75, respectively), response and remission rates were non clinically different at both time points, however in the "MINI mild-to-moderate suicidal risk" subgroup, MLN tended to be more rapidly active (remission rate at week 8-LOCF: 44.7%/MLN, 35.1%/VLF). The changes in CGI-S were also indicative of a significant improvement of the global illness severity with both treatments. RESULTS AND SAFETY/TOLERABILITY: The tolerability profile of both drugs was in line with their pharmacological activity. About 70% of patients in both groups experienced at least one adverse event (AE). In both groups, the most common AEs were nausea, dizziness, headache and hyperhidrosis, and, in the male patients, genito-urinary problems: orgasmic disorders (VLF only) and dysuria (MLN only). These AEs were mostly responsible for definitive treatment discontinuation for tolerability concerns. None of the 6 serious adverse events (SAEs) on MLN and 4 of the 8 SAEs on VLF were related to the test drug. CONCLUSION: MLN and VLF at flexible doses up to 200 mg/day globally exhibited similar efficacy and tolerability profiles in the long-term treatment of adults with MDD. PMID- 20004292 TI - [Chronic renal complications induced by lithium]. AB - INTRODUCTION: After 40 years of use of lithium in the treatment of mood disorders, the renal risks associated with the long-term exposure to lithium are better known. OBJECTIVE: This review is aimed at summarizing the information available in the literature regarding the impact of lithium on renal structure and function, the prevalence of renal abnormalities, the associated risk-factors and the strategy for their identification and management. METHOD: Articles were selected using a Medline search. The keywords were lithium, renal function, kidney, nephrotoxicity, renal insufficiency, side-effects, polyuria, diabetes insipidus and drug monitoring. RESULT: A well-recognized adverse effect of lithium exposure is the occurrence of nephrotic diabetes insipidus (NDI) resulting in polyuria and polydipsia, which occurs in 20% of the patients on long term lithium treatment. This side-effect is linked to a deficit in urine concentrating ability. Its occurrence is associated with the duration of lithium therapy. Although this effect of lithium is initially functional and may disappear if the treatment is rapidly stopped, it may become structural and permanent over time. The decision to stop lithium or to treat the NDI with amiloride is mainly based upon its functional impact. DISCUSSION: A debate has been ongoing for decades regarding whether or not the long-term use of lithium may cause slowly progressive renal failure. According to the recent literature, progressive renal failure occurs in approximately 20% of the patients on long term lithium treatment, among whom a few develop severe renal insufficiency due to lithium (possibly in conjunction with other somatic factors) in the form of interstitial nephritis. However, there is an increasing number of reports of patients requiring dialysis after long-term exposure to lithium. CONCLUSION: Current recommended strategies for minimising the renal side effects of lithium include: avoiding acute episodes of renal toxicity; monitoring serum lithium concentrations in order to achieve optimal efficacy at the lowest possible concentrations; monitoring serum creatinine levels at least on a yearly basis, with discontinuation of lithium use, discussion with a nephrologist if creatinine clearance decreases below 60 ml/mn; and the possible application of lithium into single daily dose. PMID- 20004293 TI - [Further evidence of the efficacy of the antidepressant Valdoxan (agomelatine) compared with conventional antidepressive drugs (from the ECNP held in Istanbul, 15 September 2009)]. PMID- 20004294 TI - Painting the picture of distressing somatic symptoms. PMID- 20004295 TI - Is there a better term than "medically unexplained symptoms"? PMID- 20004296 TI - Causal symptom attributions in somatoform disorder and chronic pain. AB - OBJECTIVE: Somatoform disorders (SFD) are defined by symptoms that lack medical explanation. This study examined the type and pattern of patients' causal attributions using a new semistructured interview technique METHODS: The Causal Attributions Interview allows to assess and weigh 15 common explanations of physical symptoms. Attributions given by 79 patients with SFD were compared with those obtained from 187 chronic pain patients. RESULTS: The test-retest reliabilities of the interview-elicited attributions were satisfactory to good. SFD patients attributed most of their symptoms to mental/emotional problems (46.9%) and somatic disease (41.1%), while the pain sample preferred physical overloading/exhaustion (56.1%), daily hastiness/time pressure (41.7%), somatic disease (39.6%), and weather influence (39.0%). On average, SFD patients chose 2.57 and pain patients 3.86 different attributions for each symptom. These numbers were substantially larger than those of initial spontaneous attributions. Correspondence analysis revealed underlying dimensions with three groups labeled "environmental," somatic," and "psychological/stress." While pure environmental attributions were rare (1.1%), somatic factors were chosen for 28.3% of the symptoms, psychological/stress for 22.1%, and the combination of both for 25.6%. Approximately 10% were attributed in a multicausal sense to all three groups. Depression was found to correlate positively with psychological/stress and negatively with somatic attributions. CONCLUSION: The results do not support the perspective that SFDs generally result from poor acknowledgement of emotional factors. SFD and chronic pain showed distinguishable attributional patterns. PMID- 20004297 TI - Which factors predict the persistence of DSM-IV depression, anxiety, and somatoform disorders in the medically ill three months post hospital discharge? AB - OBJECTIVE: This study sought to assess the persistence of DSM-IV depression, anxiety, and somatoform disorders in a sample of 206 medical patients 3 months after hospital discharge and to examine which baseline factors predicted the persistence of disorder. METHODS: Patients were interviewed using the Monash Interview for Liaison Psychiatry (a structured psychiatric interview for the medically ill) during admission and again at 3 months post discharge. Scales completed during admission elicited sociodemographic data, psychiatric history, mental and physical functioning, illness behavior, coping modes, and number of close relationships. Best-subset logistic regression was employed to find the best combination of these potential predictors of the persistence of psychiatric disorder. RESULTS: Persistence of anxiety disorders [n=43; 50.6%; 95% CI=39.5 61.6], depression (n=55; 44.4%; 95% CI=35.4-53.5), and somatoform disorders (n=35; 42.2%; 95% CI=31.3-53.0) was moderately high, with no statistically significant difference in the rate of persistence of the three groups of disorder. Family psychiatric history, education, and poorer physical and mental functioning during hospitalization predicted persistence of depression. Poorer mental functioning, less denial, and greater number of close relationships predicted persistence of anxiety disorders. Higher levels of education, use of acceptance-resignation as a coping mechanism, and greater hypochondriasis predicted persistence of somatoform disorders. CONCLUSION: The belief that psychiatric disorders in hospitalized medically ill patients spontaneously remit after discharge is false. A substantial proportion persist for at least 3 months. Early detection and treatment is possible and warranted. Features of the illness (poorer physical and mental health) and personal and social factors identifiable at hospital admission identify patients at risk for persistence. PMID- 20004298 TI - Mindfulness-based stress reduction for chronic pain conditions: variation in treatment outcomes and role of home meditation practice. AB - OBJECTIVE: This study compared changes in bodily pain, health-related quality of life (HRQoL), and psychological symptoms during an 8-week mindfulness-based stress reduction (MBSR) program among groups of participants with different chronic pain conditions. METHODS: From 1997-2003, a longitudinal investigation of chronic pain patients (n=133) was nested within a larger prospective cohort study of heterogeneous patients participating in MBSR at a university-based Integrative Medicine center. Measures included the Short-Form 36 Health Survey and Symptom Checklist-90-Revised. Paired t tests were used to compare pre-post changes on outcome measures. Differences in treatment effect sizes were compared as a function of chronic pain condition. Correlations were examined between outcome parameters and home meditation practice. RESULTS: Outcomes differed in significance and magnitude across common chronic pain conditions. Diagnostic subgroups of patients with arthritis, back/neck pain, or two or more comorbid pain conditions demonstrated a significant change in pain intensity and functional limitations due to pain following MBSR. Participants with arthritis showed the largest treatment effects for HRQoL and psychological distress. Patients with chronic headache/migraine experienced the smallest improvement in pain and HRQoL. Patients with fibromyalgia had the smallest improvement in psychological distress. Greater home meditation practice was associated with improvement on several outcome measures, including overall psychological distress, somatization symptoms, and self-rated health, but not pain and other quality of life scales. CONCLUSION: MBSR treatment effects on pain, HRQoL and psychological well-being vary as a function of chronic pain condition and compliance with home meditation practice. PMID- 20004299 TI - Symptoms, personality traits, and stress in people with mobile phone-related symptoms and electromagnetic hypersensitivity. AB - OBJECTIVE: Some people report symptoms that they associate with electromagnetic field (EMF) exposure. These symptoms may be related to specific EMF sources or to electrical equipment in general (perceived electromagnetic hypersensitivity, EHS). Research and clinical observations suggest a difference between mobile phone (MP)-related symptoms and EHS with respect to symptom prevalence, psychological factors, and health prognosis. This study assessed prevalence of EMF-related and EMF-nonrelated symptoms, anxiety, depression, somatization, exhaustion, and stress in people with MP-related symptoms or EHS versus a population-based sample and a control sample without EMF-related symptoms. METHODS: Forty-five participants with MP-related symptoms and 71 with EHS were compared with a population-based sample (n=106) and a control group (n=63) using self-report questionnaires. RESULTS: The EHS group reported more symptoms than the MP group, both EMF-related and EMF-nonrelated. The MP group reported a high prevalence of somatosensory symptoms, whereas the EHS group reported more neurasthenic symptoms. As to self-reported personality traits and stress, the case groups differed only on somatization and listlessness in a direct comparison. In comparison with the reference groups, the MP group showed increased levels of exhaustion and depression but not of anxiety, somatization, and stress; the EHS group showed increased levels for all of the conditions except for stress. CONCLUSION: The findings support the idea of a difference between people with symptoms related to specific EMF sources and people with general EHS with respect to symptoms and anxiety, depression, somatization, exhaustion, and stress. The differences are likely to be important in the management of patients. PMID- 20004300 TI - Resistance to extinction in an odor-20% CO2 inhalation paradigm: further evidence for a symptom learning account of multiple chemical sensitivity. AB - OBJECTIVE: Multiple chemical sensitivity (MCS) is characterized by a variety of symptoms in response to nontoxic concentrations of chemicals. To further test a laboratory model of MCS based on symptom learning, we used a stronger respiratory challenge in this study than in previous studies to induce symptoms (20% CO(2) enriched air, unconditioned stimulus). Additionally, extinction of learned symptoms was investigated. METHODS: Diluted ammonia (AM) and butyric acid (BA) served as conditioned (odor) stimuli (CSs). In a learning phase, healthy participants received three breathing trials of BA mixed with CO(2) and three trials of AM mixed with regular air, or the reversed combination. In a test phase, the same trials were administered without CO(2). Immediately after odor onset, participants rated their anxiety and expectancy to experience symptoms during that trial. Self-reported symptoms were assessed after each trial; respiratory behavior was measured throughout the experiment. RESULTS: Only participants who learned to correctly anticipate symptoms (learning phase) reported elevated symptoms in response to the CS+ odor (test phase). Anticipation was more likely correct and learned symptoms were more elevated when BA had been previously associated with CO(2). Across test trials, anticipatory anxiety diminished, but learned symptoms did not. Participants failing to learn the prearranged contingencies reported overall more symptoms and anxiety. CONCLUSIONS: Strong respiratory challenges impede extinction of learned symptoms. Conscious expectancy, which may be modulated by odor quality, determines whether learned symptoms develop in response to a specific odor or to the general context. The results further support a symptom learning account of MCS. PMID- 20004301 TI - Examining maladaptive beliefs about sleep across insomnia patient groups. AB - OBJECTIVES: Unhelpful beliefs about sleep have been linked to insomnia, and increasing one's cognitive flexibility about sleep has been linked to posttreatment sleep improvement. This study evaluated whether levels of such beliefs differ across insomnia groups and whether there are particular beliefs that differ for specific insomnia subtypes. METHODS: Participants (N=1384) were people with insomnia and good sleepers ranging from 18 to 89 years old (mean=42.6; S.D.=19.4). Data from previous studies at five insomnia clinical sites were pooled to examine responses on the Dysfunctional Beliefs and Attitudes about Sleep Scale (DBAS) across differing insomnia groups. RESULTS: Group analyses revealed that those from community-based insomnia clinics and those who are hypnotic-dependent generally had the highest levels of unhelpful sleep related beliefs. With the exception of beliefs about sleep needs (wherein only community sleep clinic patients had high scores relative to good sleepers), all insomnia groups had higher scores on the 16-item DBAS (DBAS-16) than good sleepers. A validity analysis suggested that a DBAS-16 index score of >3.8 represented the level of unhelpful beliefs associated with clinically significant insomnia, although a slightly lower cutoff may be useful for identifying an unhelpful degree of sleep-related beliefs in highly screened primary-insomnia only and medical patient groups. CONCLUSIONS: This study offers descriptive data for the use of DBAS-16 across insomnia subgroups, which will help the user understand what degree of maladaptive sleep beliefs is most strongly associated with clinically significant levels of insomnia. Results also may have implications for cognitive targeting during treatment for particular insomnia groups. PMID- 20004302 TI - From dysphonia to dysphoria: Mokken scaling shows a strong, reliable hierarchy of voice symptoms in the Voice Symptom Scale questionnaire. AB - Symptoms of hoarseness (dysphonia) are common and often associated with psychological distress. The Voice Symptom Scale (VoiSS) is a 30-item self completed questionnaire concerning voice and throat symptoms. Psychometric and clinical studies on the VoiSS show that it has good reliability and validity, and a clear factorial structure. The present article presents a further advance in voice measurement from the patient's point of view. To date, there has been no examination of whether voice-related symptoms form a hierarchy; that is, whether people who suffer voice problems progress through a reliable set of problems from mild to severe. To address this question, the technique of Mokken scaling was applied to the VoiSS in 480 patients with dysphonia. A strong and reliable Mokken scale--a symptom hierarchy--was found, which included 17 of the 30 items. This new information on dysphonia shows that voice symptoms progress from voice oriented difficulties, through practical problems, to disturbances of social relationships and mood (dysphoria). The results add information about the structured phenomenology of voice problems, further establish the relationship between voice impairment and psychosocial impairment, and suggest practical applications in the assessment of dysphonic voices. PMID- 20004303 TI - Stress profile, coping style, anxiety, depression, and gastric emptying as predictors of functional dyspepsia: a case-control study. AB - OBJECTIVE: To determine whether gastric emptying, stomach peristaltic frequencies, stress profile, depressive symptoms, anxiety, and salivary cortisol levels could predict functional dyspepsia (FD). METHODS: Magnetogastrography (MGG) was used to measure gastric emptying time and the gastric peristaltic frequencies in 15 patients with FD diagnosis and in 17 healthy volunteers. In all the participants, stress profile, anxiety, and depressive symptoms were evaluated by means of standardized questionnaires, and morning salivary samples were collected for the measurement of cortisol levels. A univariate logistic regression model was used to examine the probability of the measured variables to predict the presence of FD. RESULTS: The univariate logistic regression model showed that the half-time of gastric emptying (OR=1.16 P=.01); the subscale of stress items (OR=1.08, P=.003); negative appraisal of subscale coping strategies (OR=1.03, P=0.007); anxiety (OR=1.05, P=.01); and depression (OR=1.23, P=.02) had a significant predictive value for the presence of FD. However, by applying the stricter multiple comparison criteria, only stress, negative appraisal, and anxiety arose as predictors of FD. The FD patients compared with healthy volunteers showed significantly elevated half-time of gastric emptying (P<.0006), high scores in the subscales of stress (P<.000003), in behavior type "A" (P<.04), in coping styles (P<.008), in depression (P<.0004), and in anxiety (P<.0002). CONCLUSIONS: These findings indicate that psychosocial stress, mood symptoms, and coping style are predictors of FD. The stress shows high sensibility and specificity in the patients with FD, indicating a contribution in the etiopathogenesis of dyspepsia. PMID- 20004304 TI - The Emotional Processing Scale: scale refinement and abridgement (EPS-25). AB - OBJECTIVE: The Emotional Processing Scale (EPS) is a 38-item, eight-factor self report questionnaire designed to measure emotional processing styles and deficits. Scale development is an ongoing process and our aim was to (i) refine the scale by trying out items from a new item pool and (ii) shorten the scale to enhance its clinical and research utility. METHODS: Fifteen new items were added to the original 38-item pool. The resulting 53-item scale was administered to four groups (N=690) (mental health, healthy controls, pain patients, and general medical practice attendees). Exploratory factor analysis was used to explore the underlying factor structure. RESULTS: Maximum likelihood (ML) factor analysis was used to guide the process of item selection and scale reduction. Four of the previous eight factors remained in similar form, two of the original factors were discarded, and one new factor emerged incorporating items from two previous factors. The revised version of the scale (EPS-25) has a 25-item five-factor structure. Internal reliability was moderate to high for all five factors. CONCLUSION: The psychometric properties of the revised scale appear promising, particularly in relation to the detection of differences between diagnostic groups. PMID- 20004305 TI - Construct validity of the Chinese version of the Chalder Fatigue Scale in a Chinese community sample. AB - OBJECTIVE: This paper aims to evaluate the factorial validity of the Chinese version of the Chalder Fatigue Scale (ChCFS) using a Chinese community sample. METHODS: A total of 201 Chinese adults completed the ChCFS, the 12-item Short Form Health Survey (SF12), and the Hospital Anxiety and Depression Scale (HADS). Confirmatory factor analysis was conducted to assess the fit of a one-factor model, a two-factor correlated model, and a three-factor hierarchical model in both the 14-item and 11-item versions of ChCFS to the current data. RESULTS: ChCFS successfully replicated the original two-factor structure in the current Chinese community sample, and the 11-item version demonstrated better data-model fit than the 14-item version. The instrument possessed good internal consistency (Cronbach's alpha=.863). The convergent validity with HADS and divergent validity with SF12 were also evident. CONCLUSIONS: ChCFS is valid and reliable among Chinese adults in the general population. PMID- 20004306 TI - The proposed diagnosis of somatic symptom disorders in DSM-V: two steps forward and one step backward? PMID- 20004307 TI - The concept of comorbidity in somatoform disorder--a DSM-V alternative for the DSM-IV classification of somatoform disorder. PMID- 20004309 TI - [Syphilis, 10 years after its comeback]. PMID- 20004310 TI - [Four cases of photopheresis treatment for cutaneous lupus erythematosus refractory to standard therapy]. AB - BACKGROUND: Photopheresis is a leucopheresis procedure in which cells are photoactivated by psoralen and then irradiated by ultraviolet A. We report four cases of women with refractory cutaneous lupus erythematosus (LE) who responded to this treatment. PATIENTS AND METHODS: We treated one patient with subacute LE having a contraindication to antimalarials and to thalidomide and three patients with chronic LE (lupus panniculitis, lupus tumidus and disseminated discoid LE) refractory to treatment with hydroxychloroquine, chloroquine, thalidomide and dapsone, and also, in some cases, to oral and intravenous corticosteroids, methotrexate, colchicine, acitretine, sulfasalazine, mycophenolate mofetil and intravenous immunoglobulin. Treatment consisted of two 4-hour sessions fortnightly. Only antimalarials were continued during photopheresis. RESULTS: Photopheresis had a positive effect on all four patients. We noticed complete remission in two patients and interruption of progression followed by partial remission in the other two after a mean delay of two to three months of treatment. All treatments other than antimalarials were stopped. DISCUSSION: Photopheresis appears to be an effective treatment option in patients with cutaneous LE. Due to its high cost, it should nevertheless remain an exceptional therapeutic option restricted to patients with cutaneous LE resistant to standard therapy. PMID- 20004311 TI - [Infective endocarditis in a dermatology unit]. AB - BACKGROUND: Although often clinically suspected, infectious endocarditis (IE) is frequently difficult to diagnose with certainty. Although the dermatological signs of endocarditis can vary, they must be routinely sought where there is a suspicion of IE. The aim of this study was to reveal the diversity of clinical manifestations of IE in a dermatology unit. PATIENTS AND METHODS: This retrospective study was conducted between May 2006 and May 2007 and included all patients hospitalized in the dermatology unit in whom an IE was diagnosed according to the modified Duke criteria. RESULTS: Seven patients were included with a median age of 61 years. The reasons for hospital admission were: chronic ulcers (n=1), Sezary's syndrome (n=1), atopic dermatitis (n=1), epidermolysis bullosa acquisita (n=1) and purpura (n=1). Specific dermatological manifestations of IE included necrotic lesions on the lower limbs (n=2), purpura (n=5) and splinter haemorrhages (n=1). Blood cultures were positive in 3 cases (MSSA=2, MRSA=1). One patient had serological evidence of Coxiella burnetti IE. Cutaneous sources of IE were found in 6 cases, including acute dermohypodermitis or chronic dermatosis (3), peripheral venous catheter (n=2) and haemodialysis (n=1). Transthoracic echocardiography was negative in 6 patients, whereas transoesophageal echocardiography performed in 6 patients confirmed the diagnosis in 5 cases. The mean time to diagnosis was 21 days. Among these patients, 5 died after a mean period of 78 days. CONCLUSION: Diagnosing IE remains a clinical challenge and must be routinely considered in the presence of unusual dermatological findings such as purpura or distal necrosis, but also in patients with partially or poorly controlled chronic dermatosis, which comprise an underestimated potential source of IE. Physicians treating such patients must consider the risk of IE, especially in the event of chronic dermatosis or of an invasive cutaneous procedure involving affected skin. PMID- 20004312 TI - [Biological therapy for psoriasis: practical experience at a French dermatology unit]. AB - BACKGROUND: The course of biological therapy (BT) in clinical practice may differ markedly from treatment schedules in clinical trials. Treatment modifications and patient characteristics can affect treatment safety and efficacy. In addition, long-term results concerning the use of BT in clinical practice are lacking. OBJECTIVES: To report our experience of BT in terms of short- and long-term efficacy and safety. PATIENTS AND METHODS: The retrospectively analysed cohort consisted of psoriasis patients receiving BT between 2004 and 2008. Patients in clinical trials were excluded. Mean body surface area (BSA) and Dermatology Life Quality Index were recorded. RESULTS: Fifty-eight patients undergoing 86 courses of BT were enrolled. Thirty-three patients were treated with efalizumab, 21 with infliximab and 32 with etanercept. During the study period, 40% of patients were switched to another BT. The number of patients attaining BSA-75 at 3and 6months respectively was 38% and 75% for efalizumab, 62% and 61% for infliximab, and 36% and 61% for etanercept. After 24months of follow-up, only 33% of patients (34% of patients with efalizumab, 52% with infliximab and 22% with etanercept) were still following their initial BT, with treatment being discontinued in 52% of patients due to adverse events or treatment failure. DISCUSSION: Our study confirms the efficacy and feasibility of BT in clinical practice. However, the high frequency of BT discontinuation for adverse events or non-response led to sequential therapy using different biological treatments. PMID- 20004313 TI - [Lichen striatus with nail dystrophy in an infant]. AB - BACKGROUND: Lichen striatus is an acquired eruption that develops in a linear pattern along Blaschko's lines, most commonly on the limbs. It is most often seen in children aged between 3 and 10 years. Nail involvement is uncommon. We report the case of an infant with lichen striatus and nail involvement. CASE REPORT: A 10-month-old boy was referred for evaluation of acquired onychodystrophy on his right third and fourth fingers present for 2 months. On physical examination, lateral longitudinal ridging and splitting were seen on the right third and fourth fingernails with onychodystrophy. Linear bands of bright pink papules were seen on the inside and outside edges of the third and fourth fingers. Skin biopsy revealed a lymphocytic infiltrate in the superficial dermis with exocytosis and very few dyskeratotic cells, suggesting lichen striatus. Skin and nails lesions resolved spontaneously over 9 months. DISCUSSION: Nail involvement in lichen striatus is uncommon and is frequently associated with typical skin lesions. Thirty cases have been reported in the literature since 1941. None of the subjects affected was younger than 2-years-old. Nail involvement was more frequent on the fingers, especially the thumb. As in the present case, longitudinal ridging, splitting and thinning with onychodystrophy are common clinical features of nail involvement. The diagnosis of lichen striatus is based on the presence of skin lesions and can be confirmed by biopsy. PMID- 20004314 TI - [Cheilitis: a new manifestation of gastro-oesophageal reflux?]. AB - BACKGROUND: Gastro-oesophageal reflux (GOR) is a common disease. Extra oesophageal clinical signs associated with the disease and affecting the ENT (otitis media, rhinitis, laryngitis), pulmonary and stomatological spheres have been described for a number of years. We report three cases of cheilitis potentially attributable to GOR. CASE REPORT: Three female patients consulted for cheilitis present for several months. Screening for an infectious cause was negative, as was allergological investigation using skin tests. However, questioning revealed nocturnal breathing through the mouth in all three patients as well as symptomatic GOR in one patient. Test therapy comprising proton-pump inhibitors was proposed and after 1month symptoms had subsided completely. Gastro oesophageal endoscopy performed in the subsequent weeks revealed oesophagitis in all patients. DISCUSSION: Extra-oesophageal signs described during the course of GOR mainly affect the ENT sphere and pulmonary sphere (cough, asthma) and are doubtless due to the direct caustic action of gastric secretions on the mucus membrane. Although gastroenterological investigation (endoscopy, 24-h pHmetry) may be negative, treatment with proton-pump inhibitors results in complete resolution of symptoms. The cases we describe suggest for the first time that extra-oesophageal sites affected by GOR may include the labial semimucosa. PMID- 20004315 TI - [Large-vessel granulomatous vasculitis during the course of sarcoidosis: Takayasu's arteritis?]. AB - BACKGROUND: Vascular involvement in sarcoidosis is very rare and is characterized by preferential involvement of large vessels similar to that observed in Takayasu's disease. Distinguishing between these two diseases is often difficult and constitutes a diagnostic pitfall. The association between sarcoidosis and Takayasu's arteritis is not coincidental and a common physiopathological factor may exist; it suggests a possible aetiopathogenetic relationship between sarcoidosis and Takayasu's arteritis and casts doubt on whether this form of vasculitis is a disease in its own right or simply a syndrome caused by other diseases. CASE REPORT: We report the case of a man with a 10-year history of cutaneous and pulmonary sarcoidosis who developed ischaemia of the right upper limb evocative of Takayasu's arteritis. The patient was successfully treated with oral steroids and methotrexate. DISCUSSION: This case prompts discussion about the relationship between Takayasu's disease and sarcoidosis. Physicians should be aware of the possible occurrence of granulomatous arteritis during the course of sarcoidosis which requires a special work-up. PMID- 20004316 TI - [Sorafenib-induced multiple eruptive keratoacanthomas]. AB - BACKGROUND: Sorafenib has been approved for use in the treatment of metastatic renal cell carcinoma. Cutaneous side-effects are common, including rash, hand foot syndrome, alopecia, pruritus, dry skin and erythema. We report an original unexpected cutaneous effect: multiple keratoacanthomas. In the light of a literature review of drug-induced keratoacanthomas, we discuss the potential underlying physiopathological mechanism. CASE REPORT: Three weeks after starting treatment with sorafenib for metastatic renal cell carcinoma, a 64-year-old man developed skin lesions on the face, ears, forearms and thighs having the appearance of dome-shaped nodules with central keratotic cores. Eruptive keratoacanthomas were suspected and were in fact confirmed by histology. Thanks to effective antiangiogenic treatment and the mild discomfort of the keratoacanthomas, sorafenib could be continued. Three weeks after the end of treatment, all lesions had regressed and the patient's skin returned to normal. DISCUSSION: Although the precise aetiology is unknown, the development of eruptive keratoacanthomas is associated with impaired immunity, sun exposure, viral infection, genetic predisposition, radiation therapy and exposure to chemical carcinogens. A few cases of drug-induced keratoacanthomas have been described in the literature and certain immunosuppressant drugs have been implicated. This case suggests that the novel antineoplastic agent sorafenib has a complex activity that, in addition to tyrosine kinases inhibition, includes an immunosuppressant mechanism that can occasionally cause skin lesions. PMID- 20004317 TI - [Blasch-kolinear psoriasis revealed by infliximab therapy]. AB - BACKGROUND: Blaschko-linear psoriasis is a rare disease about which only a few publications have appeared in the literature. This form of psoriasis poses problems of differential diagnosis with regard to other forms of inflammatory Blaschko-linear dermatoses. Herein, we report an original case, the linear nature of which was revealed by treatment with infliximab. CASE REPORT: A 29-year-old man presented chronic psoriasis present for 17 years and resistant to various forms of systemic therapy. Treatment with infliximab 5mg/kg given on D1, D15 and two-monthly, thereafter resulted in practically complete resolution of all skin lesions after the fourth infusion. The only remaining lesions were psoriatic erythematous-squamous, non-pustular lesions with a Blaschko-linear pattern, limited to one side, on the left arm and left leg. These lesions persisted after 10 courses of infliximab, although no other lesions reappeared. DISCUSSION: This case was original in terms of the revelation of Blaschko-linear lesions during treatment with infliximab, despite the complete disappearance of diffuse psoriatic plaques, thus suggesting the existence in this patient of two cell populations, each having a different response to biotherapy. PMID- 20004318 TI - [Campylobacter jejuni cellulitis]. PMID- 20004319 TI - [Staphylococcal skin infections in the French armed forces: epidemiological surveillance data]. PMID- 20004320 TI - [Epidemiology of bullous pemphigoid in Guadeloupe (French West Indies)]. PMID- 20004321 TI - [A phototype-based study of frostbite in the French armed forces]. PMID- 20004322 TI - [A painful and exophytic hand lesion]. PMID- 20004323 TI - [Establishing the aetiological diagnosis of congenital ichthyosis]. PMID- 20004324 TI - [Congenital ichtyosis]. PMID- 20004325 TI - [Pigmented leg lesion]. PMID- 20004326 TI - [Focal epithelial hyperplasia]. PMID- 20004327 TI - [Primary umbilical endometriosis]. PMID- 20004328 TI - [Delayed hypersensitivity to sodium lauryl sulphate in omeprazole]. PMID- 20004329 TI - [Indications of mTOR inhibitors in liver transplantation]. PMID- 20004330 TI - Everolimus de novo in liver transplantation. AB - The safety and tolerability of everolimus has been evaluated in a randomized, phase II trial, comparing 3 doses of everolimus to a placebo, in association with cyclosporine and corticosteroids, after liver transplantation. There were no significant differences between groups in the rates of the composite end point (graft failure, biopsy-proven acute rejection, graft loss, death, or loss to follow-up) or its individual components. Although there were lower rates of treated acute rejection and mortality with the higher dosages (2 and 4 mg/day), these did not reach statistical significance. Interestingly, freedom from rejection correlated with trough blood levels of everolimus: patients with levels of 3 ng/mL or less had rejection rates 3-fold higher than patients with levels exceeding 3 ng/mL. All graft losses and most deaths were associated with typical posttransplant complications, not with study medication and not due to hepatic artery thrombosis. There were no clear dose-related differences among groups for hematology parameters. After transplantation, renal function declined to a similar extent in all 4 groups. The overall incidence of infection was comparable between groups (61-77%). Although the interpretation of the results of this trial is hampered by the small sample sizes of patient groups (about 30 in each group) and the high dropout rates (about 50%), this study suggests that everolimus is an effective immunosuppressive agent with an acceptable patient tolerance and safety profile after liver transplantation. PMID- 20004331 TI - [Minimize kidney failure in transplantation patients with proliferation signal inhibitors]. AB - Chronic renal dysfunction is a multifactorial and frequent event after organ transplantation. The measurement or the estimation of glomerular filtration rate is essential to detect early progressive renal dysfunction. Proliferation signal inhibitors are nonnephrotoxic immunosuppressive drugs which may be useful to minimize calcineurin inhibitors-related side effects through a conversion strategy. Most studies in the setting of kidney transplantation showed improvement in glomerular filtration rate as high than conversion was early. Proliferation signal inhibitors may be included quickly in new immunosuppressive regimen for liver transplanted patients with chronic renal dysfunction. PMID- 20004332 TI - [Liver transplantation for hepatocellular carcinoma and potential application of PSI]. AB - Liver transplantation is considered the best treatment of hepatocellular carcinoma but its efficacy depends on the risk of tumour recurrence. The risk of recurrence depends on tumour characteristics but is also influenced by immunosuppressive regimens. Immunosuppressants of the mTOR inhibitors family share anti-tumour properties which are already taken into account in the treatment of renal carcinoma and advanced hepatocellular carcinoma. These features make interesting their use in liver transplantation for hepatocellular carcinoma, with the following goals: to reduce the risk of post-transplant tumour recurrence, to expand the indications of liver transplantation for hepatocellular carcinoma and eventually to slow down tumour growth during the waiting period. However, to date, the potential role of mTOR inhibitors after liver transplantation for hepatocellular carcinoma is only based on small encouraging proof of concept studies. Large randomized studies are therefore required to further define the specific indications of these compounds. The demonstration of a beneficial impact of mTOR inhibitors in the setting of liver transplantation for hepatocellular carcinoma would be a major therapeutical advance. PMID- 20004333 TI - [Antitumoral effect of proliferation signal inhibitors]. AB - The mammalian target of rapamycin (mTOR) is implicated in cell growth especially during cancer development and progression. Its action is dependent on well known oncogenic pathways that regulate tumor cell growth and cell cycle progression, in response to different stimuli. Sirolimus, temsirolimus and everolimus are specific inhibitors of mTOR that have originally been characterized by their antifungal and immunosuppressive properties, but also significantly inhibit cancer cells'proliferation, invasion, and metastasis, and promote apoptosis. In addition, mTOR inhibitors display potent antiangiogenic properties by the suppression of vascular endothelial growth factor signal transduction. The antitumoral effects of mTOR inhibitors, as a monotherapy or in combination with tyrosine kinase inhibitors or usual cytotoxic agents, have been extensively suggested in preclinical studies, including animal models. In a clinical setting, preliminary reports have demonstrated that mTOR inhibitors use is associated with an acceptable safety profile. Currently, mTOR inhibitors are tested in multiple trials and various cancer types, usually in intermittent schedules to avoid significant immunosuppression. Of particular interest is the use of mTOR inhibitors in the field of organ transplantation, including liver transplantation, in preventive or curative strategies, for the treatment of recurrent hepatocellular carcinoma and de novo post-transplantation malignancies. PMID- 20004334 TI - [Side effects of proliferation signal inhibitors and their management]. AB - Proliferation signal inhibitors (PSI) could help to lower the calcineurine inhibitors level to minimize their toxicity and improve long-term graft survival. Side effects of this drugs are specific and must been known. Hyperlipidemia and cutaneous side-effects are the most frequent, angioedema and interstital pneumonitis the most serious. In majority of cases, early and adapted management could limit the impact of these side effects. PMID- 20004335 TI - Increased intake of ethanol and dietary fat in galanin overexpressing mice. AB - Evidence suggests that the orexigenic peptide, galanin (GAL), in the hypothalamic paraventricular nucleus (PVN) has a role in stimulating the consumption of ethanol, in addition to a high-fat diet. This possibility was further examined in mutant mice that overexpress the GAL gene. Two sets of GAL-overexpressors (GALOE) compared with wild-type (WT) controls, maintained on laboratory chow and water, were trained to voluntarily drink increasing concentrations of ethanol, from 3 to 15%. In the GALOE versus WT mice, the results revealed the following: (1) a 35 40% increase in ethanol intake and ethanol preference, which was evident only at the highest (15%) ethanol concentration, in male but not female mice, and was seen with comparisons to littermate and nonlittermate WT controls, (2) a significantly larger, 60-75% increase in ethanol intake and ethanol preference after a day of food deprivation, again only in male GALOE mice, (3) no change in consumption of sucrose or quinine solutions in preference tests, and (4) a 55% increase in consumption of a fat-rich diet during a 2-h test period, in both male and female GALOE mice. These results obtained with overexpression of the GAL gene provide strong support for a physiological role of this peptide in stimulating the consumption of ethanol and a fat-rich diet. They reveal gender differences in the behavioral phenotype, which may reflect GAL's functional relationship to reproductive hormones in the stimulation of consummatory behavior. PMID- 20004336 TI - Nicotinic receptor ligands reduce ethanol intake by high alcohol-drinking HAD-2 rats. AB - Neuronal nicotinic acetylcholine receptors (nAChRs) are implicated in the reinforcing effects of many drugs of abuse, including ethanol. The present study examined the efficacy of cytisine, a nAChR partial agonist, and lobeline, a putative nAChR antagonist, on the maintenance of ethanol drinking by HAD-2 rats. Adult male HAD-2 rats were given access to ethanol (15 and 30%, with ad libitum access to water and food) 22 h/day for 12 weeks, beginning at 60 days of age, after which cytisine (0.0, 0.5, and 1.5 mg/kg) was tested for 3 consecutive days. The rats were given an 18-day washout period and were then tested with lobeline (0.0, 1.0, and 5.0 mg/kg) for 3 consecutive days. Ethanol intake was measured at 1, 4, and 22 h postinjection. Rats were injected intraperitoneally just before lights out (1200 h). There was a significant main effect of cytisine treatment on the second test day, with the 1.5 mg/kg dose significantly reducing ethanol intake at the 1- and 4-h time-points, relative to saline, and the 0.5 mg/kg dose inducing a significant reduction at the 4-h time-point. Conversely, lobeline treatment resulted in significant main effects of treatment for all three time points within each test day, with the 5.0 mg/kg dose significantly reducing ethanol intake, relative to saline, at each time-point within each test day. These findings provide further evidence that activity at the nAChR influences ethanol intake and is a promising target for pharmacotherapy development for the treatment of alcohol dependence and relapse. PMID- 20004337 TI - Differential effects of free versus imposed motor activity on alcohol consumption in C57BL/6J versus DBA/2J mice. AB - Practicing sport regularly provides obvious physiological and psychological benefits, but intensive sport activity, for example, at the competition level, may be associated with an increased risk for addictive disorder, whether to improve results (doping), to tolerate consequences of important pressure and physical activities (drugs), or developing as a substitute when stopping, definitely or temporally, this activity that has been considered as potentially addictive (conversion). An animal model of voluntary versus imposed physical activity is described in which males of two mouse strains with high (C57BL/6J) versus (DBA/2J) low alcohol preference were tested for possible modifications of alcohol consumption during and after physical activity, when the activity was forced versus when it was recreative. Free access to a running wheel allowed a moderate physical activity that was associated with a decrease in voluntary alcohol intake in C57BL/6J mice. On the one hand, forced running at a higher speed than that occurring under free conditions led to increased alcohol intake, back to the initial level. On the other hand, alcohol intake remained at the same low level whether or not DBA/2J mice were subjected to free or forced running. The only effect of forced running in these alcohol-avoiding mice was a significant desynchronization of circadian motor activity rhythm, which was even more pronounced than that induced in alcohol-preferring C57BL/6J mice. These results highlight the positive aspects of moderate and entertaining sport activity and provide further support to the idea that it can decrease spontaneous alcohol intake. They also tend to demonstrate that, at least for vulnerable subjects, forced and intensive activities may not have the same benefits regarding alcohol consumption. PMID- 20004340 TI - Ethanol alters the expressions of c-Fos and myelin basic protein in differentiating oligodendrocytes. AB - Myelination occurs in the central nervous system of the human fetus, adolescents, and young adults. Ethanol interferes with myelination in part by altering the composition of the myelin sheath. Here we show that ethanol also affected the expression of the transcription factor c-Fos in differentiating oligodendrocytes (OLGs). Central glial-4 OLG progenitors were induced to differentiate in the absence and presence of 100 mM ethanol, and ethanol-caused changes in the levels of c-Fos and myelin basic protein (MBP) were determined by Western blot analysis at selected developmental stages. The relatively high c-Fos level in progenitors did not immediately decrease to a low level at the onset of differentiation but displayed a downregulation at a later developmental stage. Ethanol delayed the developmental c-Fos downregulation maintaining c-Fos at a 45% higher level at 2 days of differentiation (DoD). Ethanol also decreased the rate of the burst of MBP expression that occurred between 1 and 2 DoD, reducing the MBP level by 47% at 2 DoD. The ethanol-caused delays of c-Fos downregulation and MBP upregulation were both blocked by the protein kinase C (PKC) inhibitor bisindolylmaleimide I (BIM). Likewise, treatment of OLGs with a low 5-nM concentration of the PKC activator by 12-O-tetradecanoylphorbol-13-acetate mimicked the ethanol effects on the expression of both proteins, effects that were also counteracted by BIM. The results indicate that ethanol-caused delays of the stage-specific c-Fos downregulation and the inhibition of MBP expression both occur through a PKC mediated mechanism. The ethanol-caused delay in c-Fos downregulation may disrupt normal timing for expression of genes involved in OLG differentiation, and the inhibited MBP expression may alter the myelin sheath composition. PMID- 20004339 TI - AMPAR-mediated synaptic transmission in the CA1 hippocampal region of neonatal rats: unexpected resistance to repeated ethanol exposure. AB - Alpha-amino-3-hydroxyl-5-methyl-4-isoxazole-propionate glutamatergic receptors (AMPAR) mediate most of the fast excitatory synaptic transmission in mature neurons. In contrast, a number of developing synapses do not express AMPARs; these are gradually acquired in an activity-driven manner during the first week of life in rats, which is equivalent to the third trimester of human pregnancy. Neuronal stimulation has been shown to drive high conductance Ca(2+)-permeable AMPARs into the synapse, strengthening glutamatergic synaptic transmission. Alterations in this process could induce premature stabilization or inappropriate elimination of newly formed synapses and contribute to the hippocampal abnormalities associated with fetal alcohol spectrum disorder. Previous studies from our laboratory performed with hippocampal slices from neonatal rats showed that acute ethanol exposure exerts potent stimulant effects on CA1 and CA3 neuronal networks. However, the impact of these in vitro actions of acute ethanol exposure is unknown. Here, we tested the hypothesis that repeated in vivo exposure to ethanol strengthens AMPAR-mediated neurotransmission in the CA1 region by means of an increase in synaptic expression of Ca(2+)-permeable AMPARs. We exposed rats to ethanol vapor (serum ethanol concentration approximately 40 mM) or air for 4h/day from postnatal day (P) 2-6. In brain slices prepared at P4 6, we found no significant effect of ethanol exposure on input-output curves for AMPAR-mediated field excitatory postsynaptic potentials (fEPSPs), the contribution of Ca(2+)-permeable AMPARs to these fEPSPs, or the acute effect of ethanol on fEPSP amplitude. These results suggest that homeostatic plasticity mechanisms act to maintain glutamatergic synaptic strength and ethanol sensitivity in response to repeated developmental ethanol exposure. PMID- 20004338 TI - The effects of acute and chronic ethanol exposure on presynaptic and postsynaptic gamma-aminobutyric acid (GABA) neurotransmission in cultured cortical and hippocampal neurons. AB - Decades after ethanol was first described as a gamma-aminobutyric acid (GABA) mimetic, the precise mechanisms that produce the acute effects of ethanol and the physiological adaptations that underlie ethanol tolerance and dependence remain unclear. Although a substantial body of evidence suggests that ethanol acts on GABAergic neurotransmission to enhance inhibition in the central nervous system, the precise mechanisms underlying the physiological effects of both acute and chronic ethanol exposure are still under investigation. We have used in vitro ethanol exposure followed by recording of miniature inhibitory postsynaptic currents (mIPSCs) to determine whether acute or chronic ethanol exposure directly alters synaptic GABA(A) receptor (GABA(A)R) function or GABA release in cultured cortical and hippocampal neurons. Acute ethanol exposure slightly increased the duration of mIPSCs in hippocampal neurons but did not alter mIPSC kinetics in cortical neurons. Acute ethanol exposure did not change mIPSC frequency in either hippocampal or cortical neurons. One day of chronic ethanol exposure produced a transient decrease in mIPSC duration in cortical neurons but did not alter mIPSC kinetics in hippocampal neurons. Chronic ethanol exposure did not change mIPSC frequency in either hippocampal or cortical neurons. Chronic ethanol exposure also did not produce substantial cross-tolerance to a benzodiazepine in either hippocampal or cortical neurons. The results suggest that ethanol exposure in vitro has limited effects on synaptic GABA(A)R function and action potential independent GABA release in cultured neurons and that ethanol exposure in cultured cortical and hippocampal neurons may not reproduce all the effects that occur in vivo and in acute brain slices. PMID- 20004341 TI - Alcohol intake and 19-year mortality in diabetic men: NIPPON DATA80. AB - Although moderate alcohol intake in diabetic Caucasians is associated with a reduction in coronary heart disease mortality, no study in Japanese with diabetes has examined the association between alcohol intake and mortality outcomes. We analyzed the relationship between alcohol intake and all-cause and cause-specific mortality using the database from NIPPON DATA80. At the baseline in 1980, data on history, lifestyle, and physical examinations were collected on study participants aged 30 years and older from randomly selected areas in Japan. After excluding participants with comorbidities, we followed 4,018 male participants (3,614 nondiabetics, 195 with impaired glucose tolerance and 209 diabetic) for 19 years. During the 19 years of follow-up, there were 990 deaths; 328 were from cardiovascular disease and 157 from all-heart diseases. With the never-drinking category serving as a reference, the Cox multivariate-adjusted hazard ratios for non-daily and daily drinkers for cardiovascular mortality were 0.43 (95% confidence intervals: 0.19-0.95) and 0.45 (0.25-0.80), respectively, and 0.33 (0.12-0.91) and 0.31 (0.15-0.67) for all-heart disease mortality in the combined impaired glucose tolerance and diabetic Japanese men. Alcohol drinking in men with glucose intolerance was associated with a significant reduction in cardiovascular and all-heart disease mortality as seen in the general population in Japan. PMID- 20004342 TI - Alcohol consumption and higher incidence of impaired fasting glucose or type 2 diabetes in obese Korean men. AB - It is inconclusive whether moderate alcohol consumption reduces the diabetes risk. We observed the development of impaired fasting glucose or type 2 diabetes according to the amount of alcohol intake and body mass index. The annual health evaluation data of 2,500 male workers from 2002 to 2006 were reviewed retrospectively deleting personal identification code. The information contained sex, age, medical history, smoking status, alcohol consumption, participating regular exercise, anthropometric, and biochemistry measurement. Impaired fasting glucose or diabetes was determined when fasting plasma glucose was > or =100mg/dL. Thousand seven hundred seven subjects were eligible after excluding medical history of diabetes or fasting glucose > or =100mg/dL at baseline. The relative risks of its development in group of taking 1-14, 15-29, and > or =30.0g ethanol were 0.842 (95% confidence interval [CI], 0.603-1.176), 1.068 (95% CI, 0.736-1.551), and 1.019 (95% CI, 0.662-1.568) within normal weight group, 1.164 (95% CI, 0.795-1.705), 1.421 (95% CI, 0.947-2.133), and 1.604 (95% CI, 1.031 2.495) within overweight group, and 1.498 (95% CI, 1.042-2.153), 1.634 (95% CI, 1.091-2.447), and 1.563 (95% CI, 1.019-2.396) within obese group each after adjusting age, family history of diabetes, smoking, exercise, serum fasting glucose, aspartate aminotransferase, and gamma-glutamyltransferase with nondrinkers as a reference group. Not only high alcohol consumption but also moderate drinking was related with higher incidence of impaired fasting glucose or diabetes in obese Korean men. PMID- 20004343 TI - Alcoholism and alcohol abstinence: N-acetylcysteine to improve energy expenditure, myocardial oxidative stress, and energy metabolism in alcoholic heart disease. AB - Alcoholism has been associated with a wide range of pathologic conditions, including alcoholic heart disease (AHD). Because AHD may be associated with oxidative stress, antioxidant compounds, such as N-acetylcysteine (NAC) could be useful to control the damage done by alcohol (ethanol) consumption. To investigate the NAC effects on alcoholism and alcohol abstinence, initially, 30 male Wistar rats were divided into two groups: (C, N=6) given standard chow and water; (E, N=24) receiving standard chow and aqueous ethanol solution in semi voluntary research. After 30 days of ethanol-exposure, (E) group was divided into four subgroups (N=6/group):(E-E) continued drinking 30% ethanol-solution; (E-NAC) drinking ethanol-solution containing 2g/L NAC; (AB) changed ethanol solution to water; (AB-NAC) changed ethanol to aqueous solution of 2g/L NAC. After 15 days of the E-group division, E-E rats had lower body weight and feed efficiency, as well as higher energy-expenditure resting metabolic rate (RMR)/body weight and VO(2) consumption/surface area. These calorimetric changes were reflected on the cardiac tissue. E-E rats had higher heart weight/body weight ratio and myocardial lipid hydroperoxide (LH), indicating AHD with hypertrophy and oxidative stress. Myocardial superoxide dismutase was higher, whereas glutathione-peroxidase (GSH peroxidase) was lower in E-E rats than in C. The higher myocardial hydroxyacyl coenzyme-A dehydrogenase (OHADH), OHADH/citrate synthase (CS), and lactate dehydrogenase (LDH)/CS in E-E rats indicated higher fatty acid degradation relative to aerobic metabolism predisposing the lipotoxicity. AB rats had lower RMR/body weight than E-E, normalized myocardial oxidative stress, and energy metabolism. E-NAC and AB-NAC had lower RMR/body weight, myocardial LH, LDH/CS, and higher GSH-peroxidase than E-E and AB, respectively, demonstrating lower oxidative stress and higher myocardial carbohydrate oxidation. In conclusion, the present study brought new insights on alcohol consumption and AHD because ethanol exposure enhanced energy-expenditure and induced a number of calorimetric changes, which were reflected in body weight and myocardial lipotoxicity. NAC preventing ethanol-induced calorimetric changes and reducing myocardial oxidative stress enhanced carbohydrate oxidation, thus optimizing myocardial energy metabolism in both alcoholic and abstinence condition. PMID- 20004344 TI - Drinking and driving: a decrease in executive frontal functions in young drivers with high blood alcohol concentration. AB - This study correlated the executive frontal functions with blood alcohol concentration (BAC) in night drivers in a Brazilian city. Of 592 drivers randomly recruited between December 17, 2005 and May 5, 2006, during nighttime hours on main streets or avenues with intense vehicle traffic in Vitoria, Brazil, 444 had the BAC determined by a portable digital breath alcohol analyzer and 389 were submitted to a frontal function examination by a frontal assessment battery (FAB). A high percentage (24.4%) of drivers presented alcohol in their blood. Most of these drivers were male (82%), and nearly half (43.7%) were young adults (aged between 20 and 30 years). The results showed an inverse relationship between the BAC and FAB total scores, with a higher BAC corresponding to a smaller FAB total score, delineating a progressive decrease in frontal function with increasing concentrations of alcohol. The most intriguing result was that alcohol-induced impairment on frontal executive function was particularly important in young adults, and more specifically in the motor programming subset of FAB, an executive function highly involved in driving skills. Considering the worldwide evidence of the high-risk involvement of youth in automobile crashes, the effects of alcohol in young adults need to be more thoroughly examined by cognitive studies, and more direct preventive solutions need to be taken focusing on this age range. PMID- 20004354 TI - The vanishing physician-scientist? PMID- 20004355 TI - Stem cell therapy for the heart: a perspective. PMID- 20004356 TI - Treatment with alpha-lipoic acid reduces asymmetric dimethylarginine in patients with type 2 diabetes mellitus. AB - Elevated asymmetric dimethylarginine (ADMA) concentrations predict cardiovascular events in patients with type 2 diabetes mellitus (T2DM). It has been shown that alpha-lipoic acid (ALA) improves endothelial function and oxidative stress in these patients. The present study investigated if ALA reduces ADMA in patients with T2DM. Plasma concentrations of ADMA, L-arginine and symmetric dimethylarginine (SDMA) were determined in a double-blind, randomized, placebo controlled study in patients with T2DM. Intravenous ALA (n = 16) or placebo (n = 14) was administered daily for 3 weeks. ALA reduced ADMA while no change was observed with placebo (mean change -0.05 micromol/1[95% CI: -0.01; -0.09] vs. 0.01 micromol/1 [95% CI: -0.05; -0.03]; ANOVA p = 0.031). SDMA and L-arginine were not affected by ALA. In conclusion ALA treatment reduces ADMA in patients with T2DM. Long-term studies need to demonstrate if ALA may cause cardiovascular risk reduction. PMID- 20004357 TI - Stem cell therapy in acute myocardial infarction: a review of clinical trials. AB - Stem cells (SCs) possess the ability to differentiate into cells of various tissues. Although the differentiation of SCs into functional cardiomyocytes has been difficult to demonstrate in humans, clinical trials using SCs in the setting of acute myocardial infarction (AMI) have demonstrated variable results. Interpretation of these trials has been difficult because of multiple variables, which include differences in trial design, cell type, timing of cell delivery, and outcome measurements. Herein, a summary of all clinical trials in subgroups of direct injection, indirect mobilization, and combination approaches of SC therapy in AMI is provided with significant findings in each group. PMID- 20004358 TI - Relevance of the CX3CL1/fractalkine-CX3CR1 pathway in vasculitis and vasculopathy. AB - The clinical presentation of systemic vasculitis can vary widely and include skin disorders, neuropathy, eye symptoms, and systemic inflammation. The precise molecular mechanisms underlying this syndrome are not fully understood, but the importance of a chronic imbalance of the cytokines and chemokines involved in orchestrating inflammatory responses is now recognized. In similar fashion, atherosclerosis is now recognized to be a chronic inflammatory disease in which chemokines play important roles. In the current review, we discuss the involvement of CX3CL1, which is a unique member of the chemokine family, and its receptor, CX3CR1, in the pathogenesis of these vasculopathies. PMID- 20004359 TI - Phosphorylation of contractile proteins in response to alpha- and beta-adrenergic stimulation in neonatal cardiomyocytes. AB - alpha- and beta-Adrenergic receptor agonists induce an inotropic response in the adult heart by promoting the phosphorylation of several regulatory proteins, including myosin-binding protein C (MyBP-C), cardiac troponin I (cTnI), and phospholamban (PLB). However, the adrenergic-induced phosphorylation of these proteins has not been characterized in the developing heart. Accordingly, we evaluated MyBP-C, cTnI, and PLB phosphorylation in cultured neonatal rat cardiomyocytes (NRCMs) after alpha- and beta-receptor activation with phenylephrine and isoproterenol. alpha-Receptor stimulation increased, whereas beta-receptor activation reduced MyBP-C phosphorylation. Isoelectric-focusing experiments indicated that the amount of monophosphorylated MyBP-C was sensitive to alpha-adrenergic activation, but diphosphorylated and triphosphorylated MyBP-C levels were largely unaffected. The phosphorylation of cTnI and PLB was consistent with the mechanism observed in adult hearts: alpha- and beta-Receptor stimulation phosphorylated both proteins. For cTnI, the greatest difference associated with beta-receptor activation was observed in the diphosphorylated state, whereas alpha-receptor activation was associated with a marked increase in the tetraphosphorylated protein and absence of the unphosphorylated state. Despite these apparent changes in cTnI and PLB phosphorylation, beta-receptor activation failed to alter calcium transients in NRCMs. Collectively, these findings suggest that, unlike cTnI and PLB, MyBP-C and inotropy are not coupled to beta-adrenergic stimulation in NRCMs. Therefore, cTnI and PLB probably play a more central role in modulating contractile function in NRCMs in response to catecholamines than does MyBP-C, and MyBP-C may have a structural role in stabilizing thick filament assembly rather than influencing cross-bridge formation in developing hearts. PMID- 20004360 TI - Systemic and airway inflammation in sleep apnea and obesity: the role of ICAM-1 and IL-8. AB - The recurrent hypoxic stress that characterizes obstructive sleep apnea (OSA) seems to play a role in the increased adherence of neutrophils to endothelial cells as well as in the resulting migration of the former to the inflamed area. Intercellular adhesion molecule 1 (ICAM-1) and interleukin (IL)-8 are markers widely used in OSA studies to investigate inflammation. The aim of this study was to measure ICAM-1 and IL-8 levels in the breath condensate and in the plasma and inflammatory cells in the induced sputum of 12 obese OSA (OO) patients, 10 nonobese OSA (NOO) patients, 10 obese non-OSA (ONO) subjects, and 8 healthy subjects (HS) using a specific enzyme immunoassay (EIA) kit. A significant increase in both plasma and exhaled IL-8 and ICAM concentrations and percentage neutrophils was observed in the induced sputum of obese OSA patients, non-obese OSA patients, and obese non-OSA subjects compared with healthy subjects. However, although these inflammatory markers were found to follow an upward trend in obese OSA patients no difference was observed in both either non-obese OSA patients and obese non-OSA subjects. Finally, a significant positive correlation was found to occur among IL-8, ICAM-1, and sputum neutrophils, as well as across the apnea hypopnoea index (AHI), TST 90%, body mass index (BMI), and neck circumference. The data obtained confirm the occurrence of an ICAM- and IL-8-mediated neutrophilic airway inflammation in both OSA and obese patients. The degree of inflammation, which seems to worsen in cases of comorbidity (OSA and obesity), is likely to be responsible for the increased risk of developing cardiovascular events observed in these subjects, and therefore, it deserves to be elucidated even more. PMID- 20004362 TI - Comparison of two heights for forward-placed trunk support with standing work. AB - Two forward-placed supports with different heights are investigated using human motion capture and EMG. Ten male participants stood in 10 degrees increments of trunk flexion between 0 and 40 degrees for three conditions; leaning on a desk adjusted to the height of the pelvis, leaning on a prototype support at the height of the sternum and with no external support. Low back and hip extensor muscle activity was reduced by an average 60% with leaning against the prototype compared to the no-support condition whereas leaning on a desk produced no significant change in muscle activity. Supported conditions resulted in greater forward displacement of the trunk by at least two-fold compared to no-support representing a longer reach distance. No adverse changes in kinematics indicate that either support blocked segmental flexion of the pelvis, lumbar spine or thoracic spine. These findings suggest that leaning against a higher-placed trunk support could be beneficial for tasks requiring forward flexion. PMID- 20004361 TI - Lung endothelial barrier protection by iloprost in the 2-hit models of ventilator induced lung injury (VILI) involves inhibition of Rho signaling. AB - Mechanical ventilation at high tidal volume (HTV) may cause pulmonary capillary leakage and acute lung inflammation culminating in ventilator-induced lung injury. Iloprost is a stable, synthetic analog of prostaglandin I(2) used to treat pulmonary hypertension, which also showed endothelium-dependent antiedemagenic effects in the models of lung injury. To test the hypothesis that iloprost may attenuate lung inflammation and lung endothelial barrier disruption caused by pathologic lung distension and coagulation system component thrombin, we used cell and animal 2-hit models of ventilator-induced lung injury. Mice received a triple injection of iloprost (2 microg/kg, intravenous instillation) at 0, 40, and 80 min after the onset of HTV mechanical ventilation (30 mL/kg, 4h), combined with the administration of a thrombin receptor-activating peptide 6 (TRAP6, 3 x 10(-7)mol/mouse, intratracheal instillation). After 4h of ventilation, bronchoalveolar lavage (BAL), histologic analysis, and measurements of Evans blue accumulation in the lung tissue were performed. The effects of iloprost on endothelial barrier dysfunction were subsequently assessed in pulmonary endothelial cells (ECs) exposed to thrombin and pathologic (18%) cyclic stretch. The combination of HTV and TRAP6 enhanced the accumulation of neutrophils in BAL fluid and lung parenchyma, as well as increased the BAL protein content and endothelial permeability judged by Evans blue extravasation in the lung tissue. These effects were markedly attenuated by iloprost. The application of 18% cyclic stretch to pulmonary ECs enhanced the thrombin-induced EC paracellular gap formation and Rho-GTPase-mediated phosphorylation of regulatory myosin light chains and myosin phosphatase. Iloprost markedly inhibited the Rho-kinase-mediated site-specific phosphorylation of myosin phosphatase, and it prevented cyclic stretch- and thrombin-induced endothelial monolayer disruption. This study characterizes for the first time the protective effects of iloprost in the in vitro and in vivo 2-hit models of VILI and supports consideration of iloprost as a new therapeutic treatment of VILI. PMID- 20004363 TI - Reduced density of calbindin immunoreactive GABAergic neurons in the occipital cortex in major depression: relevance to neuroimaging studies. AB - BACKGROUND: Several lines of evidence suggest dysfunction of the gamma aminobutyric acid (GABA)ergic system in major depressive disorder. Neuroimaging studies report reduced levels of GABA in the dorsolateral prefrontal and occipital cortex of depressed patients. Our previous postmortem study revealed a reduction in the density and size of calbindin-immunoreactive (CB-IR) GABAergic neurons in the prefrontal cortex in major depressive disorder. The goal of this study was to test whether the changes in CB-IR neurons can also be detected in the occipital cortex, where neuroimaging studies report a prominent GABA decrease. METHODS: A three-dimensional cell counting probe was used to assess the cell-packing density and size of CB-IR neurons in layer II of the occipital cortex in 10 major depressive disorder subjects and 10 psychiatrically healthy control subjects. RESULTS: The density of CB-IR neurons was significantly decreased by 28% in major depressive disorder subjects compared with the control group. The size of CB-IR neurons was unchanged in major depressive disorder subjects when compared with control subjects. CONCLUSIONS: The reduction in the density of CB-IR GABAergic neurons in the occipital cortex in depression is similar to that observed previously in the prefrontal cortex. Deficit in cortical GABAergic interneurons may contribute to the low GABA levels detected in neuroimaging studies in major depressive disorder patients. PMID- 20004364 TI - Anhedonia and emotional experience in schizophrenia: neural and behavioral indicators. AB - BACKGROUND: Emotional impairments such as anhedonia are often considered key features of schizophrenia. However, self-report research suggests that emotional experience in response to affect-eliciting stimuli is intact in schizophrenia. Investigation of neural activity during emotional experience may help clarify whether symptoms of anhedonia more likely reflect alterations of in-the-moment hedonic experience or impairments in other aspects of goal-directed behavior. METHODS: Forty individuals with DSM-IV-TR schizophrenia or schizoaffective disorder and 32 healthy control subjects underwent functional magnetic resonance imaging while making valence and arousal ratings in response to emotional pictures, words, and faces. Blood oxygen level-dependent responses were compared between patients and control subjects and were correlated with questionnaire measures of anhedonia. RESULTS: Patients showed some evidence of blunted valence but not arousal ratings in response to emotional stimuli compared with control subjects. Higher anhedonia scores were associated with blunted valence ratings in both groups and fully mediated the group differences in valence ratings. Functional activity was largely intact in patients, except for regions in right ventral striatum and left putamen, which showed reduced responses to positive stimuli. Higher anhedonia was associated with reduced activation to positive versus negative stimuli in bilateral amygdala and right ventral striatum in patients and in bilateral caudate in control subjects. CONCLUSIONS: Increased anhedonia is associated with a reduced experience of valence in both patients and control subjects, and group differences in experienced valence are likely driven by individual differences in anhedonia. Reduced activation of the striatum and amygdala may contribute to symptoms of anhedonia by failing to signal the salience of positive events. PMID- 20004365 TI - Neural activity to a partner's facial expression predicts self-regulation after conflict. AB - BACKGROUND: Failure to self-regulate after an interpersonal conflict can result in persistent negative mood and maladaptive behaviors. Research indicates that lateral prefrontal cortex (LPFC) activity is related to emotion regulation in response to laboratory-based affective challenges, such as viewing emotional pictures. This suggests that compromised LPFC function may be a risk factor for mood and behavior problems after an interpersonal conflict. However, it remains unclear whether LPFC activity to a laboratory-based affective challenge predicts self-regulation in real life. METHODS: We investigated whether LPFC activity to a laboratory-based affective challenge (negative facial expressions of a partner) predicts self-regulation after a real-life affective challenge (interpersonal conflict). During a functional magnetic resonance imaging scan, healthy, adult participants in committed relationships (n = 27) viewed positive, negative, and neutral facial expressions of their partners. In a three-week online daily diary, participants reported conflict occurrence, level of negative mood, rumination, and substance use. RESULTS: LPFC activity in response to the laboratory-based affective challenge predicted self-regulation after an interpersonal conflict in daily life. When there was no interpersonal conflict, LPFC activity was not related to mood or behavior the next day. However, when an interpersonal conflict did occur, ventral LPFC (VLPFC) activity predicted mood and behavior the next day, such that lower VLPFC activity was related to higher levels of negative mood, rumination, and substance use. CONCLUSIONS: Low LPFC function may be a vulnerability and high LPFC function may be a protective factor for the development of mood and behavior problems after an interpersonal stressor. PMID- 20004366 TI - Synthesis of new glycosyl biuret and urea derivatives as potential glycoenzyme inhibitors. AB - O-peracetylated 1-(beta-D-glucopyranosyl)-5-phenylbiuret was prepared in the reaction of O-peracetylated beta-D-glucopyranosylisocyanate and phenylurea. The reaction of O-peracetylated N-beta-D-glucopyranosylurea with phenylisocyanate furnished the corresponding 1-(beta-D-glucopyranosyl)-3,5-diphenyl- as well as 3 (beta-D-glucopyranosyl)-1,5-diphenyl biurets besides 1-(beta-D-glucopyranosyl)-3 phenylurea. O-Peracetylated 1-(beta-D-glucopyranosyl)-5-(beta-D glycopyranosyl)biurets were obtained in one-pot reactions of O-peracetylated beta D-glucopyranosylamine with OCNCOCl followed by a second glycopyranosylamine of beta-D-gluco, beta-D-galacto and beta-D-xylo configurations. O-Acyl protected 1 (beta-D-glucopyranosyl)-3-(beta-D-glycopyranosylcarbonyl)ureas were obtained from the reaction of beta-D-glucopyranosylisocyanate with C-(glycopyranosyl)formamides of beta-D-gluco and beta-D-galacto configurations. The O-acyl protecting groups were removed under acid- or base-catalyzed transesterification conditions, except for the N-acylurea derivatives where the cleavage of the N-acyl groups was faster than deprotection. Some of the new compounds exhibited moderate inhibition against rabbit muscle glycogen phosphorylase b and human salivary alpha-amylase. PMID- 20004367 TI - Studies on the boronation of methyl-beta-D-cellobioside--a cellulose model. AB - The conversion of phenylboronic acid (PBA) with methyl-beta-D-cellobioside (Me beta-D-clb) and cellodextrins (DP(w) 12) was investigated to gain a basic understanding of the interactions of boric acid derivatives with oligo- and polyglucans. By means of MS and NMR experiments, it was possible to show a first stage formation of a six-membered ring at C-4 and C-6 of the non-reducing glucose occurs as in the case of monosaccharides. If the amount of reagent is increased the formation of seven-membered rings at the secondary OH moieties is observed. Even the existence of two of these large ring-systems in the direct neighborhood was found. Application of an excess of boronation reagent led to dimerization reactions of Me-beta-D-clb via the primary reducing glucose residue as confirmed by DOSY NMR studies. Preliminary (13)C NMR studies for the interaction of cellodextrins with PBA in DMSO solution confirmed a functionalization at the trans-1,2-diol moieties of these oligomers. The amount of reagent applied may either was shown to lead to soluble products or to insoluble cross-linked material. PMID- 20004368 TI - Synthesis of raphanuside, an unusual oxathiane-fused thioglucoside isolated from the seeds of Raphanus sativus L. AB - Raphanuside (1), an unusual oxathiane-fused thioglycoside isolated from the seeds of Raphanus sativus L., was synthesized via 10 steps and in 11% overall yield. PMID- 20004369 TI - Xenoreactivity of human clonal mesenchymal stem cells in a major histocompatibility complex-matched allogeneic graft-versus-host disease mouse model. AB - Effects of mesenchymal stem cells (MSCs) on graft-versus-host disease (GVHD) have been actively investigated since the discovery of the immunomodulation property of MSCs about a decade ago. Human clonal MSCs (hcMSCs) were isolated from human bone marrow aspirate according to our newly established isolation protocol called subfractionation culturing method, and were evaluated for their efficacy on GVHD treatment, using a mouse MHC-matched B6-->BALB.B GVHD model system. Although the hcMSCs can suppress the allogeneic proliferation of human peripheral blood mononuclear cells in in vitro, the administration of the hcMSCs failed to reduce the GVHD-related mortality of the murine recipients. One of the reasons might be that murine cytokines such as IFN-gamma and TNF-alpha cannot activate the hcMSCs. Based on these results, we suggest that xenogeneic MSCs may not be used for the treatment of GVHD. PMID- 20004370 TI - Effect of genistein therapy on plasma levels of asymmetric dimethylarginine in healthy postmenopausal women: a randomized, placebo-controlled study. AB - The aim of this study was to investigate the effects of pure genistein therapy on asymmetric dimethylarginine in healthy postmenopausal women. Healthy postmenopausal women received pure genistein (n=21) or placebo (n=17) for 6 months, and no statistically significant effects on plasma asymmetric dimethylarginine levels were found with pure genistein treatment. PMID- 20004371 TI - Lifestyle management improves quality of life and depression in overweight and obese women with polycystic ovary syndrome. AB - OBJECTIVE: To assess the impact of adding exercise to dietary restriction on depressive symptoms and health-related quality of life (HRQOL) in women with polycystic ovary syndrome (PCOS). DESIGN: Analysis of depression and quality of life outcomes from a randomized, controlled prospective clinical intervention that evaluated the effects on a range of health outcomes in women with PCOS. SETTING: Clinical research unit. PATIENT(S): One hundred four overweight/obese PCOS women (aged 29.3 +/- 0.7 years; body mass index [BMI] 36.1 +/- 0.5 kg/m(2)). INTERVENTION(S): Randomized to one of three 20-week lifestyle programs: diet only, diet and aerobic exercise, or diet and combined aerobic-resistance exercise. MAIN OUTCOME MEASURE(S): Depression and PCOS-specific HRQOL. RESULT(S): Forty-nine women completed the intervention (diet only = 14, diet and aerobic exercise = 15, diet and combined aerobic-resistance exercise = 20). By week 20 all groups achieved weight loss and had improvements in depression and PCOS specific HRQOL scores, except for body hair domain score. There was no difference between treatments for all outcomes. CONCLUSION(S): This study demonstrated that dietary restriction alone and combined with exercise had similar benefits for improving depression and HRQOL scores in overweight and obese women with PCOS. PMID- 20004372 TI - Endometrial preparation for in vitro maturation treatment. PMID- 20004373 TI - A novel two-step strategy for in vitro culture of early-stage ovarian follicles in the mouse. AB - OBJECTIVE: To develop an in vitro strategy to support the growth of early-stage follicles and produce mature oocytes competent for fertilization. DESIGN: Whole ovaries from 8-day-old mice were cultured for 4 days, and then secondary follicles were isolated and cultured for 12 days in a three-dimensional alginate or fibrin-alginate (FA) hydrogel matrix. SETTING: University-affiliated laboratory. ANIMALS: Mice. INTERVENTION(S): None. MAIN OUTCOME MEASURES: Histologic evaluation of follicle development, steroid hormone production, and rates of oocyte maturation, oocyte fertilization, and embryo formation. RESULT(S): Culture of 8-day-old mouse ovaries for 4 days resulted in transition of the follicle population from primordial and primary follicles to secondary follicles, similar to that seen in a 12-day-old ovary. Isolated secondary follicles cultured for 12 days showed larger increases in oocyte diameter and more frequent antrum formation and theca cell differentiation in the FA-hydrogel matrix compared with the alginate matrix. Steroid hormone secretion patterns were consistent with the changes in follicle morphology and cell differentiation observed in the cultured follicles. Compared with oocytes from alginate follicle cultures, a greater number of oocytes retrieved from the FA-based follicle cultures progressed to metaphase I, reached metaphase II, and could be fertilized and cleaved to two-cell embryos. The organ culture plus FA-hydrogel follicle culture strategy produced a very high rate of oocyte progression to metaphase II (88 +/- 8.7% [mean +/- SEM]) and formation of two-cell embryos (54 +/- 4%). CONCLUSION(S): A strategy combining whole ovary culture of early-stage follicles and subsequent FA hydrogel in vitro follicle culture produced a high percentage of oocytes competent for fertilization; this might provide new options for fertility preservation in women and prepubertal girls facing fertility threatening diseases or treatments. PMID- 20004374 TI - Vascular endothelial growth factor and beta-human chorionic gonadotropin are associated with trophoblastic invasion into the tubal wall in ectopic pregnancy. AB - OBJECTIVE: To assess the association between the depth of trophoblastic penetration into the tubal wall with serum concentrations of vascular endothelial growth factor (VEGF) and beta-hCG and to assess its predictive value. DESIGN: Prospective study. SETTING: Tertiary care university hospital. PATIENT(S): Thirty patients with ampullary pregnancy undergoing salpingectomy were analyzed. INTERVENTION(S): Trophoblastic invasion was histologically classified as stage I when limited to the tubal mucosa, stage II when extending to the muscle layer, and stage III in the case of complete tubal wall infiltration. MAIN OUTCOME MEASURE(S): The relation between depth of trophoblastic infiltration into the tubal wall with VEGF and beta-hCG serum concentrations on the day of surgery. RESULT(S): An association between the depth of trophoblastic invasion and maternal serum concentrations of VEGF and beta-hCG was observed. VEGF levels of 297.2 pg/mL showed 100.0% sensitivity and 90.0% specificity for stage I, and levels of 440.1 pg/mL showed 81.8% sensitivity and 88.8% specificity for stage III. Beta-hCG levels of 2590.0 mIU/mL showed 88.9% sensitivity and 80.0% specificity for stage I, and levels of 10,827.0 mUI/mL showed 72.7% sensitivity and 88.9% specificity for stage III. CONCLUSION(S): Maternal serum VEGF and beta hCG concentrations are associated with depth of trophoblastic penetration into the tubal wall. PMID- 20004375 TI - Cross-over design trials in infertility--how much multiplicity is too much? PMID- 20004376 TI - Tumor necrosis factor-like weak inducer of apoptosis (TWEAK)/fibroblast growth factor inducible-14 might regulate the effects of interleukin 18 and 15 in the human endometrium. AB - Tumor necrosis factor-like weak inducer of apoptosis (TWEAK) could play a role in the regulation of interleukin (IL)-15 and IL-18, cytokines crucial for angiogenesis and uterine natural killer (uNK) cell recruitment during embryo implantation. We therefore confirmed the endometrial presence of TWEAK/fibroblast growth factor inducible-14 (Fn-14) and documented simultaneously the cytotoxic KIR receptor (NKp46) of uNK cells in the human endometrium while TWEAK, Fn-14, IL 15, and IL-18 mRNA were quantified by real-time polymerase chain reaction in relation to the recruitment of CD56+ cells among fertile control women and patients who had failed to implant after assisted reproduction treatment. PMID- 20004377 TI - Pregnancy in women with polycystic ovary syndrome: the effect of different phenotypes and features on obstetric and neonatal outcomes. AB - OBJECTIVE: To test the hypothesis that the risk of adverse obstetric or neonatal outcomes varies according to different phenotypes of polycystic ovary syndrome (PCOS), and to evaluate the clinical impact of the main features of PCOS. DESIGN: Prospective controlled clinical study. SETTING: Academic Departments of Obstetrics and Gynecology, and of Endocrinology, Italy. PATIENT(S): Ninety-seven pregnant women with PCOS and 73 healthy pregnant subjects were recruited as cases and controls, respectively. INTERVENTION(S): Clinical, biochemical, and ultrasonographic evaluations. MAIN OUTCOME MEASURE(S): Obstetric and neonatal outcomes. RESULT(S): The relative risk (RR) for adverse obstetric or neonatal outcomes was increased (1.7, 95% confidence interval [CI] 1.12-2.96) in patients with PCOS and varied according to the PCOS phenotype (1.93, 95% CI 1.12-2.96; 2.23, 95% CI 1.21-3.15; 0.54, 95% CI 0.09-1.63, and 0.48, 95% CI 0.31-0.78 for full-blown, nonpolycystic ovaries [PCO], nonhyperandrogenic, and ovulatory phenotypes, respectively). The RRs were 1.57 (95% CI 0.85-2.52) and 0.48 (95% CI 0.31-0.78) for oligoanovulatory and ovulatory patients with PCOS, respectively. The risk for adverse obstetric or neonatal outcomes was affected significantly by ovarian dysfunction and biochemical hyperandrogenism, whereas no significant effect was detected for clinical hyperandrogenism and PCO. CONCLUSION(S): The increased risk for adverse obstetric and neonatal outcomes that was observed in patients with PCOS varies widely according to the different phenotypes and features of PCOS. PMID- 20004378 TI - B lymphocyte stimulator -817C>T promoter polymorphism and the predisposition for the development of deep infiltrating endometriosis. AB - The prevalence of the BLyS -817C>T polymorphic variant among women with either deep infiltrating endometriosis or adenomyosis compared with a group of gynecologic patients without symptomatic endometriosis and a group of healthy women was assessed in this study. Patients with deep infiltrating endometriosis had less often a BLyS -817C/T genotype as compared with the reference group, with an odds ratio of 0.50 (95% confidence interval 0.27-0.93 versus the C/C genotype). PMID- 20004379 TI - Sperm chromatin structure is associated with the quality of spermatogenesis in infertile patients. AB - OBJECTIVE: To establish the diagnostic value of sperm chromatin structure assessment for the evaluation of male factor infertility, in addition to conventional andrological workup. DESIGN: Cross-sectional controlled study. SETTING: A tertiary referral andrology clinic. PATIENT(S): Two hundred seventy nine male partners of infertile couples. INTERVENTION(S): None. MAIN OUTCOME MEASURE(S): The DNA fragmentation index (DFI) determined by the sperm chromatin structure assay (SCSA), semen parameters, serum levels of reproductive hormones, and World Health Organization (WHO) classification of male factor subfertility. RESULT(S): In all patient categories, except those including patients with hypogonadotrophic hypogonadism, sperm antibodies, or normospermia, DFI was significantly higher compared with in proven fertile controls. After classification of the quality of spermatogenesis based on mean testicular volume (<10 ml vs. >15 ml), follicle stimulating hormone (FSH; > 10 U/L vs. <5 U/L), and inhibin-B (<100 nmol/L vs. >150 nmol/L), the DFI was significantly higher in patients with poor spermatogenesis (35.9%) than in patients with normal spermatogenesis (25.9%). In a multiple regression analysis, the teratozoospermia index, sperm vitality, and FSH were significant determinants of the DFI level. Male age was associated with DFI, but leukocytospermia, body mass index, and smoking were not confounders of DFI. CONCLUSION(S): Impaired spermatogenesis, irrespective of the WHO classification of male factor subfertility, is generally associated with an increase of sperm DNA damage. PMID- 20004380 TI - Robotically assisted laparoscopic microsurgical tubal reanastomosis: a retrospective study. AB - OBJECTIVE: To evaluate the pregnancy and delivery outcome of robot-assisted tubal reanastomosis. DESIGN: Retrospective cohort study. SETTING: University hospital. PATIENT(S): Ninety-seven patients with available follow-up who underwent the reversal of tubal ligation, with a median age of 37 years (range, 24-47 years). INTERVENTION(S): Tubal reanastomosis by robot-assisted laparoscopy. MAIN OUTCOME MEASURE(S): Analysis of the distribution of time to conception and to estimate the crude pregnancy and birth rates at 2 years. RESULT(S): The overall pregnancy and birth rates were 71%, (95% confidence interval [CI], 61%-80%) and 62% (95% CI, 52%-72%). Ninety-one percent (95% CI, 76%-98%) of patients <35 years old became pregnant, and 88% (95% CI, 72%-97%) delivered at least once. The corresponding pregnancy and delivery rates were 75% (95% CI, 57%-89%) and 66% (95% CI, 47%-81%) between 36 and 39 years old, 50% (95% CI, 25%-75%) and 43.8% (95% CI, 20%-70%) between 40 and 42 years old, 33% (95% CI, 10%-65%) and 8.3% (95% CI, <1%-38%) after the age of 43 years. CONCLUSION(S): This study reports satisfactory birth rates after tubal reanastomosis by robot-assisted laparoscopy in patients aged 40 years or less. PMID- 20004381 TI - Prolonged time to pregnancy is associated with a greater risk of adverse outcomes. AB - Births with known time to pregnancy (TTP) during the period 1989-2007 (n=17,114) were analyzed to investigate associations between TTP and pregnancy outcome among couples that conceived spontaneously. The adjusted odds ratio (95% confidence interval) for poor neonatal health, including low Apgar score, low umbilical vein pH, and need for neonatal intensive care, was 1.51 (1.09-2.09) in women who had a TTP of 25-36 months and 1.60 (1.18-2.19) in women who had a TTP of >or=37 months compared with women with a TTP of 0-6 months. PMID- 20004385 TI - The unresolved origin of uniparental diploid cell lines. PMID- 20004387 TI - Endometriosis at several sites, cyclic bowel symptoms, and the likelihood of the appendix being affected. AB - In patients with endometriosis of the appendix, other sites are use to be affected by the disease, mainly bladder, rectosigmoid and retrocervical endometriosis. When these characteristics are present or if patients have more than three sites affected by endometriosis, the surgeon should evaluate the appendix carefully. PMID- 20004388 TI - Ovarian stimulation for in vitro fertilization alters the intrauterine cytokine, chemokine, and growth factor milieu encountered by the embryo. AB - OBJECTIVE: To elucidate the impact of ovarian stimulation on the intrauterine milieu represented by the cytokine, chemokine, and growth factor profile in endometrial secretions aspirated before embryo transfer. DESIGN: Prospective cohort study. SETTING: Fertility center in tertiary referral university hospital. PATIENT(S): Forty-two patients undergoing ovarian stimulation with GnRH analogues were recruited. They participated in both a natural and an ovarian-stimulated cycle for within patient comparisons. INTERVENTION(S): Endometrial secretion aspiration was performed immediately before embryo transfer. MAIN OUTCOME MEASURE(S): The concentrations of 17 mediators known to be involved in human embryo implantation were assessed by multiplex immunoassay. RESULT(S): After correction for multiple testing, significantly higher concentrations of interleukin (IL)-1beta, IL-5, IL-10, IL-12, IL-17, tumor necrosis factor (TNF) alpha, heparin-binding epidermal growth factor (HbEGF), eotaxin, and dickkopf homologue-1 were present in endometrial secretions obtained in stimulated compared with natural cycles. CONCLUSION(S): Endometrial secretion analysis provides a novel means of investigating the effect of ovarian stimulation on the intrauterine milieu. The in vivo milieu encountered by the embryo after transfer is significantly altered by ovarian stimulation. PMID- 20004389 TI - An aromatase polymorphism modulates the relationship between weight and estradiol levels in obese men. AB - OBJECTIVE: To describe the influence of the TTTA aromatase polymorphism (TTTAn) on the relation between obesity and plasma estradiol (E(2)) in obese men. DESIGN: A 2-year cohort study. SETTING: Clinical research center. PATIENT(S): Severely obese men (31 who had had gastric bypass surgery and 118 controls). INTERVENTION(S): Men were genotyped for the TTTAn CYP19A1 polymorphism. Anthropomorphic measures, plasma E(2), and other hormonal levels were determined at baseline and 2-year follow-up. MAIN OUTCOMES MEASURE(S): Relationships between weight and changes in weight and plasma E(2) were examined in relation to the TTTAn polymorphism. RESULT(S): The mean age was 46.5 +/- 10.82 years, and mean body mass index was 47.1 +/- 8.46 kg/m(2). The most common repeats were 7 and 11. TTTAn number did not correlate with plasma E(2) in the univariate analysis. When patients were stratified per weight group, the correlation between plasma E(2) and weight was seen only among men with a higher TTTA repeat at baseline and 2 years. Similarly, only men with higher TTTA exhibited reduced E(2) levels after weight loss. CONCLUSION(S): A higher TTTA repeat is associated with a strengthened relationship between obesity and E(2). The well-established effect of increased weight on plasma E(2) appears to be absent in men with low TTTA numbers. PMID- 20004390 TI - Effects of ovarian puncture for in vitro maturation on subsequent in vitro fertilization cycle in women with polycystic ovaries. AB - We conducted a case-control study and compared the results of IVF treatment in two groups of women with polycystic ovaries. The first group consisted of 42 women who were previously treated unsuccessfully with in vitro maturation (IVM), and the second group, 48 others with no previous IVM (control group). We included the first IVF cycle only. Our preliminary result shows that previous ovarian puncture for IVM is associated with improved number of mature oocytes retrieved and total number of embryos produced. However, it is not associated with a better pregnancy rate. A larger study is needed. PMID- 20004391 TI - Patient's fertility desire should be taken into consideration in the surgical treatment algorithm of infiltrating endometriosis. PMID- 20004392 TI - Accuracy of assisted reproductive technology information on the Massachusetts birth certificate, 1997-2000. AB - OBJECTIVE: To evaluate the accuracy of assisted reproductive technology (ART) reporting on the Massachusetts birth certificate and to explore the individual and hospital-level characteristics associated with ART reporting. DESIGN: Validity analysis of population-based data. SETTING: Live-birth deliveries by Massachusetts-resident mothers during 1997-2000. PATIENT(S): Live births and delivery mothers. INTERVENTION(S): The ART data maintained by the Centers for Disease Control and Prevention were linked with the live birth-infant death records in Massachusetts. Successfully linked records were used as the gold standard for ART-related deliveries in evaluating the validity of the ART information reported on the Massachusetts birth certificate. MAIN OUTCOME MEASURE(S): Sensitivity and specificity. RESULT(S): The sensitivity of ART reporting on the birth certificate was 27% and the specificity >99%. Sensitivity of ART reporting was higher among women with multiple deliveries (twins: 32%; triplets+: 43%) and preterm deliveries (36%). CONCLUSION(S): During the period evaluated, reporting of ART information on the birth certificate was incomplete, and ART births identified through the birth certificate were a biased sample of the population of ART births. Using delivery hospital data as the sole source of ART information for the standard birth certificate may yield inaccurate information. PMID- 20004393 TI - A diagnosis of an intraluminal carcinoid tumor of the bile duct by using cholangioscopy. PMID- 20004395 TI - Moral distress among Ugandan nurses providing HIV care: a critical ethnography. AB - BACKGROUND: The phenomenon of moral distress among nurses has been described in a variety of high-income countries and practice settings. Defined as the biopsychosocial, cognitive, and behavioural effects experienced by clinicians when their values are compromised by internal or external constraints, it results from the inability to provide the desired care to patients. No research has been reported that addresses moral distress in severely resource-challenged regions such as sub-Saharan Africa. AIM: To describe the manifestation and impact of moral distress as it was experienced by Ugandan nurses who provided care to HIV infected or -affected people. METHOD: A critical ethnography was conducted with 24 acute care and public health nurses at a large referral centre in Uganda. Data were collected through interviews, observation, and focus group discussions. RESULTS: Participants described their passion for nursing and commitment to patients. They experienced moral distress when a lack of resources put patients' wellbeing at risk. The trauma imposed by systemic challenges on the nursing profession was acknowledged, as was the perception that the public blamed nurses for poor patient outcomes. However, participants were determined to serve to the best of their abilities and to take satisfaction from any contributions they were able to make. They cited the importance of education in the development of their capacity to provide care with a positive attitude, and demonstrated a collective resilience as they discussed strategies for addressing issues that affected them and their colleagues. CONCLUSIONS: The experience of moral distress among nurses in Uganda differed somewhat from the experience of nurses in high-income countries. Constraints imposed by the inability to implement skills and knowledge to their fullest extent, as well as a lack of resources and infrastructure may result in the omission of care for patients. Moral distress appears to manifest within a relational and contextual environment and participants focussed on the impact for patients, communities, and the nursing profession as a whole, rather than on their own personal suffering. The opportunity for continuing education led to strategies to transform personal attitudes and practice as well as to enhance the presentation of the profession to the public. PMID- 20004396 TI - The user experience of critical care discharge: a meta-synthesis of qualitative research. AB - OBJECTIVES: This review identifies the most significant factors, which impact upon the user experience of progress and recovery from critical illness during the first month after discharge from critical care, and discusses these in relation to the development of effective critical care discharge support strategies. DESIGN: Meta-synthesis of qualitative primary research. DATA SOURCES: Qualitative research published in English between 1990 and 2009 was identified using online databases: CINAHL, MEDLINE, EMBASE, British Nursing Index, CDSR, ACP Journal Club, Cochrane library, Social Policy and Practice and PsycInfo. Studies of adult patients, relatives/carers/significant others, which focused on experiences after discharge from an intensive care or high dependency unit to a general ward were retrieved. REVIEW METHODS: Following screening against inclusion/exclusion criteria, methodological appraisal of studies was conducted using a published framework. Ten studies met the criteria for inclusion. RESULTS: Five key themes emerged from the meta-synthesis: physical and psychological symptoms; making progress; the need to know; and safety and security. CONCLUSIONS: Findings from this meta-synthesis and other related literature supports the existence of physical and psychological problems in the immediate period following discharge from critical care to the ward, and suggests that patients and their families have a desire for more control over their recovery. However, this desire is countered by a need to feel safe and protected, culminating in an expression of dependence on healthcare staff. Any effective support strategy needs to take account of these findings. PMID- 20004397 TI - Mechanical interaction between cells and fluid for bone tissue engineering scaffold: modulation of the interfacial shear stress. AB - An analytical model of the fluid/cell mechanical interaction was developed. The interfacial shear stress, due to the coupling between the fluid and the cell deformation, was characterized by a new dimensionless number N(fs). For N(fs) above a critical value, the fluid/cell interaction had a damping effect on the interfacial shear stress. Conversely, for N(fs) below this critical value, interfacial shear stress was amplified. As illustration, the role of the dynamic fluid/cell mechanical coupling was studied in a specific biological situation involving cells seeded in a bone scaffold. For the particular bone scaffold chosen, the dimensionless number N(fs) was higher than the critical value. In this case, the dynamic shear stress at the fluid/cell interface is damped for increasing excitation frequency. Interestingly, this damping effect is correlated to the pore diameter of the scaffold, furnishing thus target values in the design of the scaffold. Correspondingly, an efficient cell stimulation might be achieved with a scaffold of pore size larger than 300 microm as no dynamic damping effect is likely to take place. The analytical model proposed in this study, while being a simplification of a fluid/cell mechanical interaction, brings complementary insights to numerical studies by analyzing the effect of different physical parameters. PMID- 20004398 TI - Analysis of amino acids and biogenic amines in breast cancer cells by capillary electrophoresis using polymer solutions containing sodium dodecyl sulfate. AB - We describe simultaneous analysis of naphthalene-2,3-dicarboxaldehyde (NDA)-amino acid and NDA-biogenic amine derivatives by CE in conjunction with light-emitting diode-induced fluorescence detection using poly(ethylene oxide) (PEO) solutions containing sodium dodecyl sulfate (SDS). After sample injection, via EOF 0.1% PEO prepared in 100mM TB solution (pH 9.0) containing 30 mM SDS entered a capillary filled with 0.5M TB solution (pH 10.2) containing 40 mM SDS. Under this condition, 14 NDA-amino acid and NDA-amine derivatives were separated within 16 min, with high efficiency ((1.0-3.2)x10(5) theoretical plates) and sensitivity (LODs at S/N=3 ranging from 2.06 to 19.17 nM). In the presence of SDS and PEO, analytes adsorption on the capillary wall was suppressed, leading to high efficiency and reproducibility. The intraday analysis RSD values (n=3) of the mobilities for the analytes are less than 0.52%. We have validated the practicality of this approach by quantitative determination of 9 amino acids in breast cancer cells (MCF-7) and 10 amino acids in normal epithelial cells (H184B5F5/M10). The concentrations of Tau and Gln in the MCF-7 cells were different than those in the H184B5F5/M10 cells, respectively. Our results show the potential of this approach for cancer study. PMID- 20004399 TI - Properties of microdispersed sintered nanodiamonds as a stationary phase for normal-phase high performance liquid chromatography. AB - The chromatographic properties of microdispersed sintered nanodiamonds (MSND) are studied under conditions of normal-phase HPLC. The retention characteristics of 30 substances representing four classes of aromatic compounds including monoalkylbenzenes, polymethylbenzenes, di-n-alkyl phthalates and polyaromatic hydrocarbons in n-alkane mobile phases have been measured. The selectivity of MSND was compared with literature data for other common adsorbents including silica gel, alumina and porous graphitic carbon (PGC). MSND shows the distinctive adsorption properties especially in a stronger retention of aromatic hydrocarbons and in the better selectivity of the separation of geometric isomers. The significant improvement in separation efficiency up to 45,300 theoretical plates per meter, was achieved for the first time for the columns packed with diamond related materials (DRM). PMID- 20004400 TI - Systematic cyanobacterial membrane proteome analysis by combining acid hydrolysis and digestive enzymes with nano-liquid chromatography-Fourier transform mass spectrometry. AB - The identification of membrane proteins is currently under-represented since the trans-membrane domains of membrane proteins have a hydrophobic property. Membrane proteins have mainly been analyzed by cleaving and identifying exposed hydrophilic domains. We developed the membrane proteomics method for targeting integral membrane proteins by the following sequential process: in-solution acid hydrolysis, reverse phase chromatographic separation, trypsin or chymotrypsin digestion and nano-liquid chromatography-Fourier transform mass spectrometry. When we employed total membrane proteins of Synechocystis sp. PCC 6803, 155 integral membrane proteins out of a predictable 706 were identified in a single application, corresponding to 22% of a genome. The combined methods of acid hydrolysis-trypsin (AT) and acid hydrolysis-chymotrypsin (AC) identified both hydrophilic and hydrophobic domains of integral membrane proteins, respectively. The systematic approach revealed a more concrete data in mapping the repertoire of cyanobacterial membrane and membrane-linked proteome. PMID- 20004401 TI - A model for predicting slopes S in the basic equation for the linear-solvent strength theory of peptide separation by reversed-phase high-performance liquid chromatography. AB - A model for predicting the slope (S) in the fundamental equation of linear solvent-strength theory for peptidic compounds was developed. Our approach is based on the novel assumption that three well-defined molecular descriptors: peptide length (N), charge (Z) and hydrophobicity index (HI) are the major contributors to the value of S. Following the definition of the model's variables, the retention of a number of Arg-terminated synthetic peptides was investigated under isocratic elution conditions (100 A pore size C18 phase, 0.1% trifluoroacetic acid as ion-pairing modifier). The peptide sequences were systematically designed to span the properties of the typical tryptic peptides that are analyzed in proteomic experiments. Experimental data show that slopes S increase with the independent increase in both peptide charge and peptide length when the two other parameters are held constant. The influence of peptide hydrophobicity is more complex: depending on peptide length and charge, stronger RP-HPLC retention can either decrease or increase the values of S. We postulate a general function to explain this behavior: S=C1 x Z(C2)+C3 x N(C4)+C5 x HI(C6)+C7/Z+C8/N+C9/HI+C10 x ZN+C11 x ZHI+C12 x NHI+B. A simple optimization using a "random walk" through parameter-space was used to determine the optimal coefficients compared to the measured S-values of 37 peptides. The model gives a approximately 0.97 R(2) correlation between the measured and predicted S-values: it was verified against previously published data on a human growth hormone protein tryptic digest and some synthetic analogues from that mixture. PMID- 20004402 TI - The composition-explicit distillation curve technique: Relating chemical analysis and physical properties of complex fluids. AB - The analysis of complex fluids such as crude oils, fuels, vegetable oils and mixed waste streams poses significant challenges arising primarily from the multiplicity of components, the different properties of the components (polarity, polarizability, etc.) and matrix properties. We have recently introduced an analytical strategy that simplifies many of these analyses, and provides the added potential of linking compositional information with physical property information. This aspect can be used to facilitate equation of state development for the complex fluids. In addition to chemical characterization, the approach provides the ability to calculate thermodynamic properties for such complex heterogeneous streams. The technique is based on the advanced distillation curve (ADC) metrology, which separates a complex fluid by distillation into fractions that are sampled, and for which thermodynamically consistent temperatures are measured at atmospheric pressure. The collected sample fractions can be analyzed by any method that is appropriate. The analytical methods we have applied include gas chromatography (with flame ionization, mass spectrometric and sulfur chemiluminescence detection), thin layer chromatography, FTIR, corrosivity analysis, neutron activation analysis and cold neutron prompt gamma activation analysis. By far, the most widely used analytical technique we have used with the ADC is gas chromatography. This has enabled us to study finished fuels (gasoline, diesel fuels, aviation fuels, rocket propellants), crude oils (including a crude oil made from swine manure) and waste oils streams (used automotive and transformer oils). In this special issue of the Journal of Chromatography, specifically dedicated to extraction technologies, we describe the essential features of the advanced distillation curve metrology as an analytical strategy for complex fluids. PMID- 20004403 TI - Signal suppression/enhancement in high-performance liquid chromatography tandem mass spectrometry. AB - The review discusses the pitfalls of the matrix effect in mass spectrometry detection hyphenated to liquid chromatography separation. Matrix effect heavily influences both qualitative and quantitative analyses, giving rise to suppression or enhancement of the signal. As generally recognised, the predominant cause is the presence of undesired components that co-elute in the chromatographic separation and alter the ionisation process. The interfering species can be components of the sample, compounds released during the pre-treatment/extraction process or reagents added to the mobile phase to improve chromatographic resolution. The different mechanisms proposed in literature to explain the suppression or the enhancement of the signal both in electrospray and atmospheric pressure chemical ionisations are presented and the results observed in the different experimental conditions are compared and discussed. All data together lead to conclude that the chemical properties of the target analyte, the kind of matrix, the matrix to analyte concentration ratio, the extraction process, the chromatographic conditions as well as the kind of the mass spectrometry instrumentation and the ionisation conditions can play a role. Likely all these potential causes act in a synergic way and the final effect observed is hardly due to only one of them. Depending on an unpredictable combination of conditions, signal suppression or enhancement can be observed. The review discusses the matrix effects observed in HPLC-MS and HPLC-MS/MS analysis proposes hypotheses to explain the observed behaviours and proposes methods and strategies to overcome the matrix effects. PMID- 20004404 TI - Reversed-phase screening strategies for liquid chromatography on polysaccharide derived chiral stationary phases. AB - Immobilised polysaccharide-based chiral stationary phases (CSPs) are chromatographic materials that combine the remarkable enantioselective performance of the polysaccharide derivatives in addition to solvent versatility for enantiomeric resolution. Their behaviour under normal phase conditions and polar organic mode has been quite extensively discussed in several scientific communications. This article will focus on an approach to developing efficient chiral analytical methods with these immobilised CSPs (CHIRALPAK IA, CHIRALPAK IB and CHIRALPAK IC) by applying a limited number of mobile phases under reversed phase conditions. The manuscript will review the development of screening strategies by liquid chromatography for the separation of enantiomers in combination with applications compatible with LC-MS. The rational combination of this technique and the different supports will allow the identification of enantiomeric resolutions in reasonable time frames and with high success rates. PMID- 20004405 TI - Patterning of J-aggregated dyes using directed self-assembly on micro- and nanopatterned templates fabricated from phase-separated mixed Langmuir-Blodgett films. AB - We report a useful technique for the deposition of dyes in the form of J aggregates using directed self-assembly on the micro- and nanopatterned templates fabricated from the phase-separated mixed Langmuir-Blodgett (LB) films. The patterns of the mixed LB films can be tuned by adjusting the intermolecular interactions between the film-forming molecules. We used the mixed LB films containing silane coupling agent for the fabrication of micro- and nanopatterned templates, taking advantage of the difference between the surface free energy of the patterned regions and that of the self-assembled monolayers of the silane coupling agent. Atomic force microscopy showed that dyes were deposited by casting, spin-coating and the LB technique in accordance with the patterns of the original mixed LB films. Emission spectroscopy revealed that J-aggregates were formed in the patterned films. We succeeded in obtaining nanopatterns of a cyanine dye in the form of J-aggregates on the templates. PMID- 20004406 TI - One-step preparation of PS/TiO2 nanocomposite particles via miniemulsion polymerization. AB - This paper presents a one-step method to fabricate PS/TiO(2) nanocomposite particles via miniemulsion polymerization technique. In this approach, styrene (St) and acetylacetone (Acac) chelated tetra-n-butyl titanate (TBT) were confined in the miniemulsion droplets with the aid of cationic surfactant cetyl trimethyl ammonium bromide (CTAB) and co-stabilizer hexadecane (HD). During the polymerization of St, TBT diffused towards the surface of the miniemulsion droplets owing to its hydrophilicity and yielded TiO(2) particles via a sol-gel process. With the electrostatic interaction between the positively charged CTAB and negatively charged Ti-OH, TiO(2) particles were coated onto the PS cores to form PS/TiO(2) nanocomposite particles. PMID- 20004407 TI - New anthracene-tetrathiafulvalene derivative-encapsulated SWNT nanocomposite and its application for biosensing. AB - In this study, a novel anthracene-tetrathiafulvalene derivative has been synthesized and immobilized on single-walled carbon nanotubes through non covalent sidewall functionalization. The new anthracene-tetrathiafulvalene (TTF) derivative-encapsulated SWNT nanocomposites were characterized using SEM, TEM, and Raman spectra and were utilized for biomolecular recognition. Our observations demonstrate that the new anthracene-TTF derivative-encapsulated SWNT nanocomposites can readily facilitate the biosensing and sensitive detection of DNA, which could be further explored for promising applications in bioelectronics and biosensors. PMID- 20004408 TI - Cell surface characterisation of Microcystis aeruginosa and Chlorella vulgaris. AB - According to previous studies, Microcystis aeruginosa, a picocyanobacteria, is more sensitive to copper than the other phytoplanktonic species such as Chlorella vulgaris a chlorophycea. An experimental study was carried out to determine the cell wall characteristics of both species. The cell surface of M. aeruginosa presented a lower hydrophobic character and a more negative surface charge over a larger pH range than for C. vulgaris, according to hydrophobicity and zeta potentials measurements. By combining infrared spectroscopy data and potentiometric titrations, the total concentration of functional groups determined for M. aeruginosa (1.60x10(-3)mol/g), as well as the number of carboxyl groups, was higher than for the C. vulgaris ones (0.22x10(-3)mol/g). The differences in surface properties, as shown by chemical, physicochemical and spectroscopic measurements would justify the differences in sensitivities to copper ions up-take between the two specimens. PMID- 20004409 TI - Surface reactivity of alpha-Al(2)O(3) and mechanisms of phosphate sorption: In situ ATR-FTIR spectroscopy and zeta potential studies. AB - We have investigated the effect of solution parameters on the adsorption of phosphate ions and on charges and structures, i.e., on the nature of species, at the alpha-Al(2)O(3) colloid/solution interface by using the batch method, zeta potential measurements, and in situ ATR-FTIR spectroscopy. The uptake of phosphate decreases with the extent of surface deprotonation (i.e., pH), imparts negative charges to the colloid surface, and induces IEP shifts showing chemical sorption. Use of complementary techniques provides evidence that phosphate is sorbed at low pH (3.3) by a combination of surface reactions of complexation and precipitation, whose relative contributions depend on phosphate loading. Surface complexation includes fast reactions of ligand exchange with single coordinated hydroxyls, and electrostatic attraction of H(2)PO(4)(-) ions at positively charged surface sites. This is supported by experiments at low coverage showing sharp and linear decrease of zeta potential (i.e., surface charge) with amount of phosphate sorbed. At high coverage, zeta potential values are low and independent of phosphate loading. Formation of surface precipitates of Al-phosphate is inferred from the assignment of the ATR-FTIR absorption band at 1137cm(-1), whose intensity increases with phosphate solution content and reaction time, to the P-O stretching vibration mode for phosphate sorbed at high concentrations on alpha Al(2)O(3). In situ ATR-FTIR spectroscopy reveals also structural reorganizations of surface hydroxyls with time, due to surface hydration and to surface precipitation continuing over extended periods along alumina dissolution. PMID- 20004410 TI - Self-assembly of lyotropic liquid crystal phases in ternary systems of 1,2 dimethyl-3-hexadecylimidazolium bromide/1-decanol/water. AB - Ternary mixtures of 1,2-dimethyl-3-hexadecylimidazolium bromide/1-decanol/water form two types of well-ordered lyotropic ionic liquid crystals, a hexagonal phase linked to the ionic liquid-water axis, and a lamellar phase located at the center of their phase diagram. The lamellar and hexagonal phases span a wide temperature range from 3 to 40 degrees C, depending on the composition. The spontaneous self assembly of the mixtures arises from strong association between the hydroxyl groups of 1-decanol or water and the polar head-groups of the ionic liquid (head group solvent), aromatic stacking interaction between the imidazolium rings and the hydrophobic effect. PMID- 20004411 TI - Aggregation behavior of a chiral long-chain ionic liquid in aqueous solution. AB - The synthesis of a chiral long-chain ionic liquid (IL), S-3-hexadecyl-1-(1 hydroxy-propan-2-yl)-imidazolium bromide ([C(16)hpim]Br), is presented. The adsorption and aggregation of this surface active IL in aqueous solution is described. The critical micelle concentration (cmc) measurement suggests that the chiral IL has superior capacity for micelle formation compared to traditional ionic surfactants. The relatively larger hydrophilic head group of the IL results in a larger maximum surface excess concentration (Gamma(max)) and a smaller minimum molecular cross-sectional area (A(min)). Electrical conductivity studies show a small degree of counterion binding to these micelles, which may increase the electrostatic repulsions between the hydrophilic heads of adjacent surfactant molecules. Both factors of the hydrophilic headgroup size and electrostatic repulsion in [C(16)hpim]Br micelles lead to a looser packing of the surfactant molecules in the micelles. As a result, a higher micropolarity and smaller mean aggregation number is observed. Moreover, the looser micellar packing of the [C(16)hpim]Br molecules results in a unusual upfield shift of the proton NMR signals in the hydrophobic chains after micelle formation. (1)H NMR and 2D ROESY spectroscopic analyses confirm a chiral arrangement of the micelles. Chiral IL micelles may have potential applications in the stereochemical recognition of surfaces or of biological structures. PMID- 20004412 TI - A longitudinal investigation of morpho-syntax in children with Speech Sound Disorders. AB - PURPOSE: The intent of this study was to examine the longitudinal morpho syntactic progression of children with Speech Sound Disorders (SSD) grouped according to Mean Length of Utterance (MLU) scores. METHODS: Thirty-seven children separated into four clusters were assessed in their pre-kindergarten and Grade 1 years. Cluster 1 were children with typical development; the other clusters were children with SSD. Cluster 2 had good pre-kindergarten MLU; Clusters 3 and 4 had low MLU scores in pre-kindergarten, and (respectively) good and poor MLU outcomes. RESULTS: Children with SSD in pre-kindergarten had lower Developmental Sentence Scores (DSS) and made fewer attempts at finite embedded clauses than children with typical development. All children with SSD, especially Cluster 4, had difficulty with finite verb morphology. CONCLUSIONS: Children with SSD and typical MLU may be weak in some areas of syntax. Children with SSD who have low MLU scores and poor finite verb morphology skills in pre-kindergarten may be at risk for poor expressive language outcomes. However, these results need to be replicated with larger groups. LEARNING OUTCOMES: The reader should (1) have a general understanding of findings from studies on morpho-syntax and SSD conducted over the last half century (2) be aware of some potential areas of morpho-syntactic weakness in young children with SSD who nonetheless have typical MLU, and (3) be aware of some potential longitudinal predictors of continued language difficulty in young children with SSD and poor MLU. PMID- 20004413 TI - Calculating Cornell voltage from nonstandard chest electrode recording site in the Reasons for Geographic And Racial Differences in Stroke study. AB - BACKGROUND: To minimize participants' burden and the need for disrobing, a 7-lead electrocardiogram (ECG) recording using a single mid-sternal chest lead was recorded at the initial stages of The REasons for Geographic And Racial Differences in Stroke (REGARDS) study. Electrocardiogram-detected left ventricular hypertrophy (ECG-LVH) by Cornell voltage (RaVL + S-wave amplitude in V(3) [SV(3)]) cannot be assessed from this method because of the absence of V(3). We examined the possibility that the S-wave amplitude in the mid-sternal lead (SV) could be used as a surrogate for SV(3). METHODS: The REGARDS study is a US national study where 7-lead ECGs were performed in 8,330 (29%) participants and standard 12-lead EGCs were performed in 20 811 (71%). Cornell voltage was calculated as the sum of aVL amplitude + SV (in the 7-lead group) or SV(3) (in the 12-lead group). Logistic regression analysis was used to examine and compare the magnitude of the association between the LVH risk factors with ECG-LVH in both groups, and Cox proportional hazards analysis was used to examine and compare the hazard ratios of overall mortality and cardiovascular mortality associated with ECG-LVH in both groups. RESULTS: Regardless of the Cornell voltage calculation method, ECG-LVH was significantly associated with LVH risk factors; and with the exception of sex, there was no evidence of a difference in the magnitude of the association. ECG-LVH from both approaches were significantly and similarly associated with both all-cause and cardiovascular mortality. CONCLUSION: ECG-LVH by Cornell voltage calculated from a 7-lead ECG (using SV in the formula) has demographic and clinical associations that are similar to that calculated from a standard 12-lead ECG (using SV(3)). In epidemiologic studies recording 7-lead ECG, SV could be used as an alternative to SV(3) in the Cornell voltage formula. PMID- 20004414 TI - Presence of soil-dwelling clostridia in commercial powdered infant formulas. AB - OBJECTIVE: Because Clostridium botulinum was isolated from powdered infant formula (PIF) fed to an infant in the United Kingdom who subsequently developed infant botulism and from unopened PIF from the same manufacturer, we tested PIF manufactured in the United States for the presence of clostridial spores. STUDY DESIGN: Thirty PIF ingested by 19 California infants with botulism within 4 weeks of onset of illness (48% of all patients fed PIF during study) in 2006-2007 were cultured anaerobically to isolate clostridia. All isolated clostridia were identified to the species level and enumerated with standard microbiologic and molecular methods. RESULTS: Five of 30 (17%) PIF samples ingested by patients contained clostridial spores. Spores were also found in 7 of 9 (78%) market purchased PIF samples. Clostridium sporogenes was isolated most frequently, followed by Clostridium butyricum and at least 10 other soil-dwelling clostridial species. No neurotoxigenic clostridia were isolated. The most probable number of clostridial spores in PIF ranged between 1.1 to >23 per 100 g. CONCLUSIONS: With the notable exception of production of botulinum neurotoxin, C sporogenes is physiologically comparable with proteolytic strains of C botulinum, and both share the same natural reservoir (soils and dust worldwide). The isolation of C sporogenes and potentially pathogenic clostridia from U.S.-manufactured PIF suggests that neurotoxigenic clostridial spores have the potential to be present in these products. PMID- 20004415 TI - Serotonergic functioning as measured by the loudness dependence of auditory evoked potentials is related to a haplotype in the brain-derived neurotrophic factor (BDNF) gene. AB - OBJECTIVES: The serotonergic system plays an important pathophysiological role in various psychiatric disorders. Brain-derived neurotrophic factor (BDNF) is involved in the differentiation and survival of serotonergic neurons. A previous study showed that low serum BDNF levels were associated with strong loudness dependence of auditory evoked potentials (LDAEP) as a reflection of low central serotonergic activity. To evaluate the genetic basis of this relationship, we studied whether the LDAEP is correlated with genetic variants within the BDNF gene. METHODS: Ninety five healthy subjects (41 males, 54 females) received electrophysiological recording of LDAEP and blood drawing for BDNF genotyping. Three BDNF markers (including the single nucleotide polymorphism rs6265(Val66Met)) were analyzed. RESULTS: Haplotype analysis revealed stronger LDAEP values in carriers of the G(Val)-C-T [rs6265(Val66Met)-rs2030324-rs1491850] haplotype within the BDNF gene in comparison to other haplotype carriers. These findings were demonstrated for the LDAEP of both left and right primary auditory cortices as well as for the vertex electrode (Cz). CONCLUSION: Subjects with the BDNF haplotype G(Val)-C-T seem to be characterized by low serotonergic activity as well as possibly by low serum BDNF levels. These findings need replication in independent samples. PMID- 20004416 TI - Baseline of butyltin pollution in coastal sediments within the Basque Country (northern Spain), in 2007-2008. AB - Tributyltin (TBT), dibutyltin (DBT) and monobutyltin (MBT) were measured in surficial sediments at, the ports of Pasaia and Bilbao, together with other mid- and small-size harbours of the Basque Country (northern Spain), in 2007-2008. The highest values of the sum of the three measured butyltin species (3523-3640 ng g( 1), as Sn) were found at sampling stations near to shipyards located within the port of Pasaia. The highest value of TBT concentration (3143 ng g(-1), as Sn) was found at the marina of Getxo, in the port of Bilbao. The degree of TBT degradation varied greatly between sampling stations, being found to be generally higher in those sediments with higher values of redox potential and lower values of TBT concentration (normalized by organic matter content). PMID- 20004417 TI - Using blood samples to estimate persistent organic pollutants and metals in green sea turtles (Chelonia mydas). AB - Persistent organic pollutants (POPs) and heavy metals have been reported in a number of green turtle (Chelonia mydas) populations worldwide. However, due to ethical considerations, these studies have generally been on tissues from deceased and stranded animals. The purpose of this study was to investigate the use of blood samples to estimate the tissue contamination of live C. mydas populations. This study analysed 125 POP compounds and eight heavy metals in the blood, liver, kidney and muscle of 16 C. mydas from the Sea World Sea Turtle Rehabilitation Program, Gold Coast, Australia. Strong correlations were observed between blood and tissue concentrations for a number of POPs and metals. Furthermore, these correlations were observed over large ranges of turtle size, sex and condition. These results indicate that blood samples are a reliable non lethal method for predicting chemical contamination in C. mydas. PMID- 20004418 TI - The role of biogenic structures on the biogeochemical functioning of mangrove constructed wetlands sediments--a mesocosm approach. AB - Benthic metabolism (measured as CO(2) production) and carbon oxidation pathways were evaluated in 4 mangrove mesocosms subjected daily to seawater or 60% sewage in the absence or presence of mangrove trees and biogenic structures (pneumatophores and crab burrows). Total CO(2) emission from darkened sediments devoid of biogenic structures at pristine conditions was comparable during inundation (immersion) and air exposure (emersion), although increased 2-7 times in sewage contaminated mesocosms. Biogenic structures increased low tide carbon gas emissions at contaminated (30%) and particularly pristine conditions (60%). When sewage was loaded into the mesocosms under unvegetated and planted conditions, iron reduction was substituted by sulfate reduction and contribution of aerobic respiration to total metabolism remained above 50%. Our results clearly show impacts of sewage on the partitioning of electron acceptors in mangrove sediment and confirm the importance of biogenic structures for biogeochemical functioning but also on greenhouse gases emission. PMID- 20004419 TI - [M129V PRNP gene polymorphism in Castilla y Leon shows a similar distribution to other Spanish regions and other European countries]. AB - BACKGROUND AND OBJECTIVE: It has been reported that the polymorphism M129V in the PRNP gene modifies the phenotype in all the subtypes of prion diseases in humans. All cases diagnosed to date as suffering the human variant of mad cow disease (vCJD) carry the allele 129M. In the last five years five cases of vCJD have been reported in Spain, three of them in Castilla y Leon (in western Spain). Observation of differences in the genotypic frequency of this polymorphism in different populations prompted us to analyze the distribution of genotypes of the V129M polymorphism in a population from Castilla y Leon to determine if there is an increased risk to develop vCJD in this region of Spain. PATIENTS AND METHOD: We studied the coding region of the PRNP gene containing codon 129 in 110 healthy individuals from Castilla y Leon. We amplified the DNA by PCR and allelic discrimination was performed after digestion with restriction enzyme NspI. Comparison between groups was performed by X(2) test. RESULTS: Our results show a distribution of genotypes of the V129M polymorphism in the population from Castilla y Leon which is similar to other Spanish and European populations. CONCLUSIONS: From the genetic point of view, the risk of people from Castilla y Leon to suffer vCJD should be similar to that of the other Spanish regions. PMID- 20004420 TI - [Current treatment of chronic hepatitis C. Factors influencing the rate of relapse and their effect on sustained virological response]. PMID- 20004421 TI - [Neotanal screening of congenital hypothyroidism. A major contribution to public health in the last few decades]. PMID- 20004422 TI - [Disseminated abdominal hydatid disease]. PMID- 20004423 TI - [The mediterranean diet: a group of healthy foods, a type of diet, or an advertising panacea?]. PMID- 20004424 TI - Hemoglobin concentration in men with type 2 diabetes mellitus. AB - Anemia is a common but often overlooked complication of diabetes. We investigated the relationship between hemoglobin concentration and various factors as well as markers of subclinical atherosclerosis in men with type 2 diabetes mellitus. Hemoglobin concentration was measured in 319 men with type 2 diabetes mellitus. We evaluated the relationship between hemoglobin concentration and various factors including age, body mass index, and glycemic control, as well as between hemoglobin concentration and pulse wave velocity or ankle-brachial index (n = 209) and between hemoglobin concentration and carotid intima-media thickness or plaque score (n = 125). Mean hemoglobin concentration was 14.2 +/- 0.80 g/dL. Body mass index (r = 0.340, P < .0001) and estimated glomerular filtration rate (r = 0.219, P = .0011) were positively associated with hemoglobin concentration, whereas age (r = -0.388, P < .0001), glycated albumin (r = -0.148, P = .0121), serum creatinine concentration (r = -0.206, P = .0019), and log (urinary albumin excretion) (r = -0.188, P = .0010) were negatively associated with hemoglobin concentration. Multiple regression analysis identified age (beta = -0.222, P = .0019), body mass index (beta = 0.145, P = .0432), systolic blood pressure (beta = 0.214, P = .0015), total cholesterol concentration (beta = 0.170, P = .0077), and serum creatinine concentration (beta = -0.181, P = .0045) as independent determinants of hemoglobin concentration. No significant association was observed between hemoglobin concentration and serum erythropoietin concentration (r = 0.079, P = .2980). Negative correlations were found between hemoglobin concentration and pulse wave velocity (r = -0.289, P < .0001) and between hemoglobin concentration and plaque score (r = -0.275, P = .0024). In conclusion, hemoglobin concentration was associated with various factors; and decreased hemoglobin concentration was associated with subclinical markers of atherosclerosis in men with type 2 diabetes mellitus. PMID- 20004425 TI - Insulin resistance is associated with C-reactive protein independent of abdominal obesity in nondiabetic Taiwanese. AB - Insulin resistance, which plays a fundamental role in the pathogenesis of metabolic syndrome and type 2 diabetes mellitus, is associated with serum levels of inflammatory markers and abdominal obesity. Whether insulin resistance is caused by inflammation or is an epiphenomenon of obesity remains unresolved. We therefore conducted a cross-sectional study to investigate whether the association between insulin resistance and C-reactive protein (CRP) levels is independent of abdominal obesity in a nondiabetic Taiwanese population. The study included 574 Taiwanese participants (300 men and 274 women) who were nondiabetic persons with CRP levels not exceeding 10 mg/L and who did not have a history of cardiovascular disease or were taking medication for dyslipidemia. All participants were of Han-Chinese origin. The degree of insulin resistance was determined using the homeostasis model assessment of insulin resistance (HOMA IR). The CRP levels were categorized into quartiles from the lowest to the highest concentrations (Q1-Q4). Blood pressure, fasting glucose level, triglycerides level, waist circumference, and HOMA-IR were all found to be significantly higher in Q3 and Q4 than in Q1 and Q2. Stratified analysis by sex and abdominal obesity showed that HOMA-IR was significantly associated with CRP levels in both sexes in either obese or nonobese populations. Multiple linear regression analysis adjusting for age, smoking, components of metabolic syndrome, and waist circumference showed that the association between HOMA-IR and CRP levels remained significant in both men and women (P = .029 for men and P < .001 for women). These findings confirm that insulin resistance is strongly associated with CRP levels independent of abdominal obesity in nondiabetic Taiwanese. Factors other than abdominal obesity, such as polymorphisms in the CRP gene, may influence the association of insulin resistance with CRP levels in different ethnic populations. PMID- 20004426 TI - Diabetic retinopathy is associated with visceral fat accumulation in Japanese type 2 diabetes mellitus patients. AB - The presence of diabetic retinopathy (DR) and increased of visceral fat accumulation (VFA) are associated with high mortality in type 2 diabetes mellitus patients. This preliminary study was therefore designed to test the hypothesis that DR is associated with insulin resistance and VFA in type 2 diabetes mellitus patients without insulin treatment. A total of 102 type 2 diabetes mellitus patients were divided into 2 groups: DR group (age, 60 +/- 6 years [mean +/- SD]; n = 31) and no diabetic retinopathy (NDR) group (59 +/- 5 years, n = 71). The level of blood glucose was assessed by fasting plasma glucose, fasting immunoreactive insulin, homeostasis model assessment index, and hemoglobin A(1c). The fat distribution was evaluated by measuring the VFA by abdominal computed tomography at the umbilical level. The body mass index and waist circumference were higher in the DR group than in the NDR group (P < .001 and P < .0005, respectively). Plasma levels of triglyceride were higher, whereas high-density lipoprotein cholesterol was lower, in the DR group than in the NDR group (P < .005 and P < .0001, respectively). Fasting plasma glucose (P < .0005), insulin concentrations (P < .0001), homeostasis model assessment index (P < .0001), and VFA (P < .0001) levels were higher in the DR group than in the NDR group. Multivariate logistic analysis revealed that DR was independently predicted by high VFA and insulin resistance. The results of this preliminary study indicate that the presence of DR was associated with high VFA and insulin resistance in Japanese patients with type 2 diabetes mellitus. PMID- 20004427 TI - Soluble adhesion molecule E-selectin predicts cardiovascular events in Japanese patients with type 2 diabetes mellitus. AB - Soluble adhesion molecule E-selectin (sE-selectin) is a marker of endothelial activation. To investigate whether high serum concentrations of sE-selectin could predict cardiovascular events, we followed 392 Japanese patients with type 2 diabetes mellitus who had no history of cardiovascular disease for a mean period of 6 years. The cardiovascular end points were defined as fatal and nonfatal myocardial infarction, angina pectoris, stroke, and sudden death. During the follow-up period, 51 patients reached end point. Patients who reached end point were significantly older and had longer duration of diabetes, higher systolic blood pressure, higher hemoglobin A1c, higher plasma glucose, higher sE-selectin, and lower high-density lipoprotein cholesterol compared with those free of such events. The mean serum concentration of sE-selectin was higher in patients who reached end point (81.1 +/- 32.2 ng/mL) than event-free patients (66.7 +/- 33.7 ng/mL, mean +/- SD; P < .01). Multiple logistic regression analysis identified age, systolic blood pressure, total cholesterol, sE-selectin, and low high density lipoprotein cholesterol as independent factors related to cardiovascular events. The odds ratio for cardiovascular events for 1-SD increase in sE-selectin concentration was 1.45 (95% confidence interval, 1.22-1.71). Kaplan-Meier analysis demonstrated a significantly higher cardiovascular event rate in the highest tertile of sE-selectin compared with the lowest or middle tertile of sE selectin (P < .01). The results suggest that high serum concentrations of sE selectin can predict cardiovascular events in Japanese patients with type 2 diabetes mellitus. PMID- 20004428 TI - Pharmacokinetics, plasma protein binding and bioavailability of moxifloxacin in Muscovy ducks after different routes of administration. AB - In this study the disposition kinetics and plasma availability of moxifloxacin in Muscovy ducks after single intravenous (i.v.), intramuscular (i.m.) and oral (p.o.) administrations of 5 mg kg(-1) b.wt. were investigated. The concentrations of moxifloxacin in the plasma were measured using high-performance liquid chromatography (HPLC) with fluorescence detection on samples collected at frequent intervals after drug administration. Following intravenous injection, the decline in plasma drug concentration was bi-exponential with half-lives of (t(1/2)(alpha)) 0.22+/-0.10h and (t(1/2)(beta)) 2.49+/-0.26h for distribution and elimination phases, respectively. The volume of distribution at steady-state (V(dss)) was 1.02+/-0.14 l kg(-1) and the total body clearance (Cl(tot)) was 0.32+/-0.11 l kg(-1)h(-1), respectively. After intramuscular and oral administration of moxifloxacin at the same dose the peak plasma concentrations (C(max)) were 2.38+/-0.43 and 2.11+/-0.36 microg ml(-1) and were obtained at 1.47+/-0.26 and 1.83+/-0.16h (T(max)), respectively, the elimination half-lives (T(1/2el)) were 3.14+/-0.42 and 2.63+/-0.44h, respectively, and AUC(0-24) were 15.87+/-2.35 and 14.52+/-2.37 microg ml(-1)h(-1), respectively. The systemic bioavailabilities were 96.36+/-11.54% and 86.79+/-12.64%, respectively. In vitro plasma protein binding percent was 32%. We concluded that moxifloxacin might be clinically interesting alternative for the treatment of most sensitive bacterial infections in Muscovy ducks. PMID- 20004429 TI - [Cerebral venous thrombosis: look for hyperthyroidism]. PMID- 20004431 TI - The relevance of transition zones on computed tomography in the management of small bowel obstruction. AB - BACKGROUND: Frequently, radiologists emphasize radiographic transition zones (RTZs) on computed tomography (CT), which are areas of abrupt change from dilated to collapsed bowel, as pathognomonic for small-bowel obstruction (SBO) diagnosis and location. The relevance of RTZs to patient management remains unknown. The purpose of this study was to determine the surgical predictive value and intraoperative accuracy of RTZ. METHODS: A retrospective review of 200 patients with SBO who underwent abdominal CT at a single institution from 2002 to 2007 was performed. Statistical analysis was conducted using an unpaired t test, a Chi square test, and multivariate analysis. RESULTS: Of the 200 patients with SBO, 150 (75%) had an RTZ. Seventy-five (38%) patients required operative intervention; 58 (39%) patients had RTZ and 17 (34%) patients did not have RTZ (P=NS). The presence of RTZ was not associated with increased probability of operative versus nonoperative management (odds ratio=1.19; 95% confidence interval [0.61-2.32]). The mean time to operative intervention was 3.6 days. Immediate operative intervention (<24 h) was equivalent in patients with versus without RTZ (57% vs 53%; P=NS) as was intervention for failed nonoperative management (43% vs 47%; P=NS). For patients who required operative intervention, RTZ correlated with intraoperative site of obstruction in only 31 (63%) patients. CONCLUSION: The presence of RTZs does not increase the likelihood of operative intervention or identify patients who will fail nonoperative management. RTZ should, therefore, not be used as a major criterion influencing operative versus nonoperative management decisions in patients with SBO. For patients who required operative intervention, RTZ had a 63% correlation with intra-operative findings, which makes it a useful adjunct to pre-operative planning. PMID- 20004430 TI - Roux-en-Y gastric bypass reverses renal glomerular but not tubular abnormalities in excessively obese diabetics. AB - BACKGROUND: Obesity and type 2 diabetes are associated with renal dysfunction, which improves after Roux-en-Y gastric bypass (RYGB). During a 12-month follow-up period, we studied prospectively the changes in glomerular and tubular functions that occurred in excessively obese diabetic and non diabetic subjects after RYGB. METHODS: The cohort included 35 patients, 54% of whom had type 2 diabetes. Glomerular filtration rate (GFR) was estimated using creatinine clearance. Tubular function was studied by measuring the ratio of urinary cystatin C to urinary creatinine (UCC ratio). RESULTS: Baseline renal parameters, anthropometric characteristics, and changes in body mass index after the surgical procedures were similar between the 2 cohorts. At 12 months after RYGB, creatinine clearance decreased 15% in diabetics (P = .02) and 21% in nondiabetics (P = .03). A change in GFR was seen earlier in the nondiabetics (-29% after 6 months; P = .003). The UCC ratio was increased at both 6- and 12-month follow-ups (P = .03 and .003, respectively) only in the diabetic group. CONCLUSION: GFR was improved at 12 months after RYGB, with nondiabetics showing a greater propensity score. Tubular function remained unchanged in the nondiabetic subjects, but worsening occurred in the diabetic subjects. These results underscore the importance of reversal of excessive obesity before the onset of frank diabetes. PMID- 20004432 TI - Matrix metalloproteinase-9 delays wound healing in a murine wound model. AB - BACKGROUND: Metalloproteinase-9 (MMP-9) is a type IV collagenase found at elevated levels in chronic wounds. As wounds heal, MMP-9 diminishes. In this study, we investigated whether MMP-9 directly contributes to chronic wound pathogenesis. METHODS: Recombinant proMMP-9 was prepared using immortalized keratinocytes transduced by a lentivirus. ProMMP-9 was purified from cell culture media and activated using 4-aminophenylmercuric acetate. Active MMP-9 was then suspended in xanthan gum to a concentration paralleling that found in human chronic wounds. Two parallel 6-mm punch biopsies were made on the backs of C57BL mice. Wounds were treated daily with MMP-9 or vehicle. Wound areas were measured and tissues examined by densitometry, real-time RT-PCR, histology, and immunohistochemistry at days 7, 10, and 12. RESULTS: Exogenous MMP-9, at the level found within chronic wounds, delayed wound healing in this animal model. By 7 days, wounds in the MMP-9-injected group were 12% larger than control wounds (P = .008). By day 12, wounds in the MMP-9-injected group were 25% larger than those of the control group (P = .03). Histologic examination shows that high levels of active MMP-9-impaired epithelial migrating tongues (P = .0008). Moreover, consistent with elevated MMP-9, the collagen IV in the leading edge of the epithelial tongue was diminished. CONCLUSION: MMP-9 appears to directly delay wound healing. Our data suggests that this may occur through interference with re epithelialization. We propose that MMP-9 interferes with the basement membrane protein structure, which in turn impedes keratinocyte migration, attachment, and the reestablishment of the epidermis. PMID- 20004433 TI - Innominate artery aneurysm as presentation of angiosarcoma. PMID- 20004434 TI - Antiangiogenic response after 70% hepatectomy and its relationship with hepatic regeneration and angiogenesis in rats. AB - BACKGROUND: The aim of this study was to evaluate the antiangiogenic response and its relation to regeneration and angiogenesis after 70% hepatectomy in a rat model. METHODS: Sixty-four Wistar albino rats were included in the study. Animals were allocated into 8 groups (n = 8). After a 70% hepatectomy, liver regeneration, angiogenesis, and antiangiogenic response were evaluated in the remnant liver on days 0, 1, 2, 3, 5, 7, 10, and 14. Regeneration and angiogenesis were determined with immunoreactivity to proliferating cell nuclear antigen and vascular endothelial growth factor. Antiangiogenic response was evaluated by detecting collagen 18 m RNA with reverse transcriptase polymerase chain reaction. RESULTS: We showed that liver regeneration peaked at day 1, whereas angiogenesis in the periportal and perisinusoidal areas reached their peak values on days 3 and 7, respectively. Both regeneration and angiogenic activity around perisinusoidal hepatocytes returned to basal activity on the day 10. Antiangiogenic response first appeared on day 5, reached a peak on day 10, and returned to basal values on day 14. CONCLUSION: Collagen18 mRNA expression is present in the normal liver during the regenerative process. We suggest that the stimulus that causes the cessation of regeneration process may come from hepatocytes, and collagen 18 produced by hepatocytes may modulate this event by inhibiting the angiogenesis. PMID- 20004435 TI - Dysphagia lucoria: consideration for the endocrine surgeon. PMID- 20004436 TI - Extended central pancreatic resection as an alternative for extended left or extended right resection for appropriate pancreatic neoplasms. AB - BACKGROUND: Whether patients with focal pancreatic lesions of benign or borderline pathology should be treated by extended central pancreatectomy rather than by extended classic resectional procedures, such as extended right and left resections, is controversial. METHODS: Between 1992 and 2007, 105 patients underwent operation for focal pancreatic lesions of borderline or benign neuroendocrine neoplasms, cystadenoma, intraductal papillary mucinous neoplasia (IPMN), and secondary metastasis. In all, 35 patients were subjected to extended central pancreatectomy, whereas the remaining 70 patients were treated by an extended classic right resection or an extended classic left resection. Groups were matched according to age, sex, and histopathology. RESULTS: No peri operative mortality occurred after extended central pancreatectomy and extended classic left resection (n = 35, each). Two (6%) patients died after extended classic right resection. Overall, in-hospital morbidity was 26% after extended central pancreatectomy, 43% after extended classic right resection, and 37% after extended classic left resection. After a median follow-up of 48 months, a local recurrence rate of 17% after extended central pancreatectomy was similar to the corresponding rates of 9% after extended classic left resection and 14% after extended classic right resection. Endocrine and exocrine impairment was less pronounced after extended central pancreatectomy (6% and 9%) than after extended classic left resection (34% and 29%) and extended classic right resection (28% and 24%; P < .05). CONCLUSION: Extended central pancreatectomy for appropriate pancreatic neoplasms is associated with less peri-operative morbidity and mortality than after extended classic left and extended classic right resection. Long-term local recurrence after extended central pancreatectomy is similar to the recurrence rates after extended classic right and classic left resection. Our results suggest that appropriately selected patients will benefit from extended central pancreatectomy because of the maintenance of endocrine and exocrine function. PMID- 20004437 TI - CEACAM-7: a predictive marker for rectal cancer recurrence. AB - BACKGROUND: The identification of rectal cancer patients predisposed to developing recurrent disease could allow directed adjuvant therapy to improve outcomes while decreasing unnecessary morbidity. This study evaluates carcinoembryonic antigen cellular adhesion molecule-7 (CEACAM-7) expression in rectal cancer as a predictive recurrence factor. METHODS: A single-institution colorectal cancer database and a frozen tissue biobank were queried for rectal cancer patients. CEACAM-7 messenger RNA (mRNA) expression from normal rectal mucosa and rectal cancers was analyzed using quantitative real-time polymerase chain reaction (PCR). Expression-level differences among normal tissue, disease free survivors, and those that developed recurrence were analyzed. RESULTS: Eighty-four patients were included in the study, which consisted of 37 patients with nonrecurrent disease (median follow-up, 170 months), 29 patients with recurrent disease, and 18 patients with stage IV disease. CEACAM-7 expression was decreased 21-fold in rectal cancers compared with normal mucosa (P = .002). The expression levels of CEACAM-7 were relatively decreased in tumors that developed recurrence compared with nonrecurrence, significantly for stage II patients (14 fold relative decrease, P = .002). For stages I-III, disease-free survival segregates were based on relative CEACAM-7 expression values (P = .036), specifically for stage II (P = .018). CONCLUSION: CEACAM-7 expression is significantly decreased in rectal cancer. Expression differences between long term survivors and those with recurrent disease introduce a potential tumor marker to define a subset of patients who benefit most from adjuvant therapy. PMID- 20004438 TI - The efficacy of balloon-occluded retrograde transvenous obliteration on small intestinal variceal bleeding. AB - BACKGROUND: Small intestinal variceal bleeding is an unusual cause of gastrointestinal hemorrhaging of portal hypertensive patients, but once it occurs it may lead to life-threatening bleeding because this problem is difficult to diagnose and treat. This study investigated the efficacy of balloon occluded retrograde transvenous obliteration (B-RTO) to treat small intestinal variceal bleeding. METHODS: From 2003 to 2007, 6 patients with small variceal bleeding were treated by B-RTO. The characteristics of the patients, B-RTO procedures, prognosis, and occurrence of risky esophageal and gastric varices were evaluated according to their medical records. RESULTS: Three-dimensional (3-D) angiography by MD-CT was useful for the diagnosis of small intestinal varices and the draining vessels. In all cases, variceal bleeding was controlled by B-RTO treatment, although rebleeding was recognized in one patient after 30 months (16.6%). All patients were alive throughout the follow-up except 1 patient who died of liver failure (6-44 months). Risky esophageal varices and gastric varices occurred in 2 patients (33.3%) after B-RTO. However, no variceal bleeding occurred. CONCLUSION: B-RTO was found to be an effective treatment modality which provided good initial hemostasis, thereby eradicating ectopic small intestinal varices. PMID- 20004439 TI - Pre-operative dental brushing can reduce the risk of postoperative pneumonia in esophageal cancer patients. AB - BACKGROUND: The presence of pathogens in dental plaque is a risk factor associated with postoperative pneumonia in esophageal cancer patients. The effectiveness of pre-operative dental brushing to decrease the risk of postoperative pneumonia in esophageal cancer patients was evaluated prospectively. METHODS: A total of 86 thoracic esophageal cancer patients who underwent an esophagectomy were investigated. Patients were divided into 2 groups: the control group (41 patients) and the pre-operative dental brushing group (45 patients). The patients in the brushing group were assigned to brush their teeth 5 times a day. After the operation, the frequency of postoperative pneumonia and need for tracheostomy for pulmonary treatment was calculated. RESULTS: Postoperative pneumonia was decreased markedly from 32% to 9% (P = .013), and the frequency of postoperative pneumonia requiring tracheostomy decreased from 12% to 0% in the dental brushing group, respectively. Limiting the patients who had positive pathogenic bacteria in their dental plaque on their admission, the frequency of postoperative pneumonia was decreased from 71% (5 of 7 patients) in the control group to 17% (2 of 12 patients) in the dental brushing group (P = .045). CONCLUSION: Frequent pre-operative dental brushing is performed easily and seems to prevent postoperative pneumonia in esophageal cancer patients. PMID- 20004441 TI - Minor versus major hepatic resection for small hepatocellular carcinoma (HCC) in cirrhotic patients: a 20-year experience. AB - BACKGROUND: The choice between minor versus major resection or anatomic versus nonantatomic resection for small (<5 cm) solitary hepatocellular carcinoma (HCC) in patients with cirrhosis is controversial. The aim of our study was to evaluate the long-term disease-free survival (DFS) and overall survival (OS) after minor or major hepatic resection for small solitary HCC in cirrhotic patients. METHODS: Between January 1983 and December 2002, patients with solitary HCC of < or = 5 cm in size who had histologically proven liver cirrhosis and microscopically tumor free margin were included. These selected patients underwent either minor (< or = 2 segments) or major (> or = 3 segments) hepatectomy. RESULTS: In 373 patients, 259 underwent minor and 114 underwent major hepatectomy. Patients in the minor resection group had more severe underlying liver disease (P = .005). Therefore, only 29.3% received anatomic resection in the minor resection group in comparison with 72.8% in the major hepatectomy group (P = .0001). No difference was found in postoperative morbidity (P = .105), mortality (P =.222), intrahepatic recurrence (P = .742), and 5-year DFS and OS (31.6% vs 31.8%, P = .932 and 50.7% vs 44.0%, P = .114) in both groups. The type of operative resection was not found to be a significant factor affecting survival in univariate analysis, but the preoperative liver function (alanine aminotransferase [AST] or alanine aminotransferase [ALT], serum albumin, or Child-Pugh status), tumor characteristics (alpha-feto protein, size, and presence of daughter nodules), and blood transfusion were found to be independent factors that affect the DFS and OS in a multivariate analysis. CONCLUSION: The severity of cirrhosis and tumor characteristics depicts long-term survival rather than the type of resection in HCC. PMID- 20004440 TI - Operative outcome of colon interposition in the treatment of esophageal cancer: a 20-year experience. AB - BACKGROUND: In patients with esophageal cancer and a history of gastric surgery, colonic interposition is the treatment of choice. Our aim was to review our experience with this technique and to identify possible predictors of the clinical outcome. METHODS: Between 1986 and 2006, 43 patients underwent esophageal reconstruction accomplished by colon interposition in our surgical department. Data from these patients were collected consecutively and reviewed retrospectively. RESULTS: Colon interposition was performed isoperistaltically in 15 patients and was performed in 28 patients anisoperistaltically. In 18 patients, the right colon was used for interposition, whereas in 25 patients, the left colon was used. The mean survival time was 23+/-29 months. Artificial ventilation more than 24h, tumor differentiation grade III, the presence of major complications, and the presence of multivisceral resection had a significant negative influence on the operative outcome of colon interposition for esophageal replacement. CONCLUSION: Colon interposition for esophageal replacement provides a satisfactory operative outcome with high complication rates. Therefore, it should be reserved as a treatment of second choice for cases in which the stomach is not available. PMID- 20004442 TI - Global survey of factors influencing choice of surgical journal for manuscript submission. AB - BACKGROUND: An increasing number of general and affiliated specialty society journals make finding the right place for manuscript submission of an article challenging. Little is known about what factors surgeons hold important when choosing a journal for article submission. MATERIALS: A global e-mail survey of authors publishing in 5 general surgery journals (Annals of Surgery, British Journal of Surgery, World Journal of Surgery, Archives of Surgery, and Surgery) from January 1, 2007, to December 31, 2008. Demographic data were collected. 15 arbitrarily chosen factors associated with submission strategy were rated for importance on a 5-point modified Likert scale (ranging from 1 representing "unimportant" and 5 representing "very important"). RESULTS: Of 1,855 authors, 250 (14%) responded. Representing 41 countries, 23 (10%) of the respondents were female and 250 (90%) were male. About two thirds of the authors had less than 10 years of clinical practice, with general surgery or gastrointestinal surgery as the major fields of interest represented. Of the 15 factors, the journal "reputation" was rated "very important" (5 points) by 62% of the respondents, followed by the journal "impact factor," which was rated "very important" by 61%, although some geographic differences were noted in this rating. Grouping several factors together in categories, the journal "prestige" and "turnaround time" category was held to be most important based on the average scores received. Age correlated with valued importance of the journal reputation (Spearman rho=0.141; P=.033). The factors considered the least important included the journal's acceptance/rejection rate, the option to suggest peer reviewers, and open access. CONCLUSION: The majority of seasoned surgeons held the overall reputation of the journal as the most important factor followed by the impact factor when choosing a journal for manuscript submission. PMID- 20004443 TI - Improving surgical outcomes through adoption of evidence-based process measures: intervention specific or associated with overall hospital quality? AB - BACKGROUND: The Leapfrog Group aims to improve surgical outcomes through promoting hospital adoption of procedure-specific process measures, although it is unclear whether compliance reflects a hospital's overall quality. The purpose of this study was to evaluate whether implementation of Leapfrog's standard for routine beta-blockade was associated with reductions in mortality after open abdominal aortic aneurysm (AAA) repair alone versus other high-risk operations. METHODS: Using a 2:1 matched case-control study design, hospitals that had not adopted the beta-blockade standard (n = 72) were compared with hospitals that had implemented this Leapfrog standard (n = 36). Leapfrog survey data were linked to patient outcomes in the California OSHPD database from 2000 to 2005. Random effects Poisson regression models were used to evaluate in-hospital mortality over time for patients undergoing AAA repair versus esophagectomy, hepatectomy, pancreatectomy, colectomy, gastrectomy, and pulmonary lobectomy. RESULTS: A total of 6,199 AAA repairs, 2,780 esophagectomies, 2,544 hepatectomies, 2,909 pancreatectomies, 57,795 colectomies, 6,267 gastrectomies, and 10,210 lobectomies were analyzed. AAA-associated mortality significantly declined in hospitals that adopted the beta-blocker standard (relative risk [RR]: 0.49; 95% confidence interval [CI]: 0.24-0.97; P < .05). Implementation of this Leapfrog standard had no effect on reducing adjusted mortality rates for other high-risk operations, including esophagectomy (RR: 0.70; 95% CI: 0.25-1.89), hepatectomy (RR: 1.16; 95% CI: 0.32-4.29), pancreatectomy (RR: 0.76; 95% CI: 0.28-2.02), colectomy (RR: 1.12; 95% CI: 0.86-1.44), gastrectomy (RR: 1.17; 95% CI: 0.57-2.43), and lobectomy (RR: 0.98; 95% CI: 0.46-2.08) (all P > .05). CONCLUSION: Compliance with peri-operative beta-blockade resulted in a significant reduction in mortality after open AAA repair over time, but it had no crossover effect on mortality associated with other high-risk operations in the same hospital. These data suggest that improvements in outcomes resulting from the adoption of evidence-based process measures are procedure specific and do not necessarily reflect overall hospital quality. PMID- 20004444 TI - Perineal reconstruction after rectal and anal disease using the local fascio cutaneous-infragluteal flap: A new and reliable technique. AB - BACKGROUND: Significant morbidity can result from perineal wounds, particularly if the tissue has been partially devitalized after radiotherapy and extensive resection for cancer or chronic inflammation which may occur in Crohn's disease. Many different types of flaps have been used to improve healing of perineal tissue defects. The purpose of this study was to evaluate the morbidity and outcomes of reconstruction using the local fasciocutaneous infragluteal (FCI) flap. METHODS: Fourteen consecutive patients undergoing local FCI flap reconstruction for perineal wounds and defects were included in the study. In 5 female patients, the defect included the dorsal wall of the vagina, which was reconstructed in 1 step. Ten patients underwent operations for anal or low rectal cancer, 3 suffered from Crohn's disease and extensive local fistula formation, and 1 young girl presented with a defect after resection of a perineal synovial sarcoma. Eleven of these patients underwent preoperative chemotherapy and either pre- or intra-operative radiotherapy. RESULTS: Complete healing of perineal wounds occurred in 13 of the 14 patients. There were 4 flap-related complications, including 3 patients with delayed wound healing and wound dehiscence and 1 patient with partial flap necrosis. The last patient required a second local flap for wound closure. In 2 patients, a second FCI flap was necessary because of a second tumor and a local tumor recurrence. Ambulation and normal mobility were possible after the flap procedure in all patients without restriction of activity. Four patients died during the follow-up period (median, 42.5 months) from tumor metastasis. CONCLUSION: Local FCI flap for reconstruction of large perineal defects achieves good wound healing with only moderate morbidity in most patients after extensive resection owing to cancer or Crohn's disease. PMID- 20004445 TI - Restricted peri-operative fluid administration adjusted by serum lactate level improved outcome after major elective surgery for gastrointestinal malignancy. AB - BACKGROUND: Our objective was to compare the effect of a restricted intravenous fluid regimen adjusted by serum lactate level with a standard restricted regimen on complications after major elective surgery for gastrointestinal malignancy. METHODS: This is a randomized, observer-blinded, single-center trial conducted across a time span of 13 months. A total of 299 patients were allocated to either a restricted intravenous fluid regimen with supplementary intravenous fluids given based on serum lactate level (group A) or a standard restricted regimen (group R). In group A, the serum lactate level was monitored closely postoperatively to maintain a normal pre-operative serum lactate level. Group R involved patients treated with a restricted fluid regimen in whom additional fluid and electrolytes were administered when deemed necessary based on the usual clinical criteria. The primary outcome measure was complications; the secondary measures were death and adverse effects. RESULTS: Additional fluid supplementation was needed in some patients in both groups (group A [28%] vs group R [26%]). In group A, the time for additional fluid infusion occurred earlier in the postoperative period than group R. Patients in group A received their first supplementary fluid treatment within the first 12 h more commonly than those in group R (74% vs 37%, respectively; P < .004). The regimen adjusted by serum lactate decreased systemic postoperative complications in group A versus group R (10% vs 22%, respectively; P = .023) but not overall total complications (23% vs 33%, respectively; P = .090). In contrast, in patients who required additional fluid infusion, the difference in complications between the 2 groups was greater (overall complication, 45% vs 85%, respectively; P = .023; major complication, 16% vs 44%, respectively; P = .018; systemic complications, 19% vs 63%, respectively; P = .001). One patient died in group A and 4 died in group R (1% vs 4%, respectively; P = .206). CONCLUSION: A fluid-restricted regimen after elective gastrointestinal operations for malignancy may lead to fluid insufficiency and low tissue perfusion in up to 28% of patients. Close monitoring of serum lactate levels with adjustment of intravenous fluid administration intraoperatively and in the early postoperative period may improve the early detection and correction of inadequate tissue perfusion, thereby decreasing the rate of complications. PMID- 20004446 TI - Adenovirus-mediated siRNA targeting Mcl-1 gene increases radiosensitivity of pancreatic carcinoma cells in vitro and in vivo. AB - BACKGROUND: Myeloid cell leukemia-1 (Mcl-1), an anti-apoptotic member of the B cell lymphoma/leukemia-2 (Bcl-2) family, has been shown to be involved in apoptosis and the cell cycle. Mcl-1 is overexpressed in many malignancies, including pancreatic cancer. The aim of this study was to investigate the effect of siRNA targeted against Mcl-1 on the radiosensitivity of human pancreatic carcinoma cells. METHODS: In 3 pancreatic cancer cell lines, the expression of Mcl-1 mRNA was determined by RT-PCR assay, and the dose-dependent cytotoxicity of radiation was also assessed. Furthermore, the effects of adenovirus-mediated siRNA targeted against Mcl-1 on radiosensitivity of PANC-1 cells both in vitro and in vivo were evaluated. RESULTS: Pancreatic cancer cells showed various levels of Mcl-1 mRNA that were correlated with the radiosensitivity of tumor cells. AdU6/shMcl-1 significantly downregulated the expression of Mcl-1 gene in PANC-1 cells, the most radioresistant cell line. Furthermore, Mcl-1 downregulation could significantly enhance radiosensitivity of PANC-1 cells in vitro and in vivo. The mechanism might be correlated with apoptosis enhancement by activating celluar caspase-3. CONCLUSION: Mcl-1 might be a therapeutic target for radiosensitization of pancreatic carcinoma cells. Adenovirus-mediated siRNA targeting of Mcl-1 could enhance the radiosensitivity of pancreatic cancer cells both in vitro and in vivo, and thus might be a potential strategy for ameliorating cancer. PMID- 20004447 TI - Pediatric small bowel intussusception disease: feasibility of screening for surgery with early computed tomographic evaluation. AB - BACKGROUND: This study investigated the feasibility of early computed tomographic (CT) evaluation and the operative results of pediatric small bowel intussusception with deteriorating ischemic or obstructive symptoms, so-called small bowel intussusception disease (SBID). METHODS: Between 1988 and 1999, among 18 patients surgically proven SBID (conventional group), 12 mimicked ileocolic intussusception and were conventionally managed with abdominal radiography, ultrasonography, reduction enema, and eventually operation. Between 2000 and 2008, we applied a modified approach with inclusion of early CT evaluation if ultrasonography showed a target lesion suspicious for SBID (diameter 25 and <35, HbA1C >7.5%) before and at 1, 4, 12, 26, and 52 weeks after sleeve gastrectomy. The insulin secretion was measured by insulinogenic index and area under the curve (AUC) during a standard oral glucose tolerance test (OGTT). Remission of type 2 diabetes was defined as fasting glucose level <126 mg/dL and HbA1C <6.5% without any glycemic therapy. RESULTS: Of the 20 patients enrolled, the mean age was 46.3 + or - 8.0 years, mean BMI was 31.0 + or - 2.9 kg/m(2), and mean HbA1C was 10.1 + or - 2.2. The mean BMI and excess body weight loss at 1, 4, 12, 26, and 52 weeks after operation were 28.9 (22.1%), 27.4 (43.0%), 25.7 (55.1%), 24.9 (71.9%), and 24.6 (69.1%), respectively. The mean HbA1C at 1, 4, 12, 26, and 52 weeks after operation were 9.2, 8.4, 7.7, 7.3, and 7.1, respectively. Resolution of type 2 diabetes was achieved in 2 (20%) patients at 4 weeks, 6 (30%) at 12 weeks, 8 (40%) at 26 weeks, and 10 (50%) at 52 weeks after sleeve gastrectomy. Before operation, the mean fasting plasma glucose and insulin levels were 240.1 + 80.9 mg/dL and 16.8 + or - 15.4 uIU/mL, respectively. The OGTT test showed a blunted insulin secretion pattern with an AUC of 3,135 uIU x min/mL. At 1 week after operation, the fasting plasma glucose and insulin levels significantly decreased to 158 + or - 52 mg/dL and 5.6 + or - 3.2 uIU/mL, respectively. The AUC decreased to 2,988.7 uIU x min/mL. The AUC at 4, 12, 26, and 52 weeks after operation was 2,211, 1,584, 3,621, and 3,351 uIU x min/mL, respectively. The diabetes resolution rates for those with pre-operative C peptide <3, 3-6, and >6 ng/mL were 1/7 (14.3%), 7/11 (63.6%), and 2/2 (100%), respectively (P < .05). CONCLUSION: Laparosopic gastric sleeve gastrectomy resulted in remission of poorly controlled nonmorbidly obese T2DM patients up to 50% at 1 year after operation. The effect is related more to the decreasing of insulin resistance because of calorie restriction and weight loss rather than to the increasing of insulin secretion. C-peptide >3 ng/mL is the most important predictor for a successful treatment. PMID- 20004452 TI - A novel treatment for bile duct injury with a tissue-engineered bioabsorbable polymer patch. AB - BACKGROUND: With the recent widespread use of laparoscopic cholecystectomy and living-donor liver transplantation, complications involving the biliary system, and stenosis in particular, are encountered frequently. Although various invasive and noninvasive techniques are now available for the treatment of biliary stenosis, recurrence and other problems limit their value and utility. Our group sought to investigate whether a bioabsorbable polymer (BAP) patch could serve as a novel treatment for biliary stenosis. METHODS: In anesthetized hybrid pigs (n = 12), a spindle-shaped portion of the lower common bile duct wall was excised, and a BAP patch was implanted at the excision site. The animals underwent repeat laparotomy at 5 weeks (n = 6) and at 4 months (n = 6) after implantation to recover the graft sites for gross and histologic studies. Blood chemistry was analyzed from samples taken during the patch implantation and recovery. RESULTS: All of the recipient pigs survived until killing. All gained weight and showed no evidence of jaundice. The BAP-patched duct remained patent without obstruction at 5 weeks postimplantation. Blood chemistry did not reveal any increases in hepatobiliary enzyme activities. Histology showed accessory glandular structures in the neo-bile duct. At 4 months, the graft site was indistinguishable from the native duct. Intra-operative cholangiography revealed dilation of the patched site, but no dilatation of the intrahepatic bile ducts. Blood chemistry values were within normal ranges. Histology at the site of the patch confirmed the growth of a cuboidal columnar epithelium similar to that of the native duct. CONCLUSION: The bile duct was dilated only focally at the site of implantation. This newly designed substitute has potential for application as a novel treatment for biliary injury and stenosis. PMID- 20004453 TI - An organized response to resident work-hours restrictions. PMID- 20004454 TI - Timing-dependent protection of hypertonic saline solution administration in experimental liver ischemia/reperfusion injury. AB - BACKGROUND: During liver ischemia, the decrease in mitochondrial energy causes cellular damage that is aggravated after reperfusion. This injury can trigger a systemic inflammatory syndrome, also producing remote organ damage. Several substances have been employed to decrease this inflammatory response during liver transplantation, liver resections, and hypovolemic shock. The aim of this study was to evaluate the effects of hypertonic saline solution and the best timing of administration to prevent organ injury during experimental liver ischemia/reperfusion. METHODS: Rats underwent 1 hr of warm liver ischemia followed by reperfusion. Eighty-four rats were allocated into 6 groups: sham group, control of ischemia group (C), pre-ischemia treated NaCl 0.9% (ISS) and NaCl 7.5% (HTS) groups, pre-reperfusion ISS, and HTS groups. Blood and tissue samples were collected 4 hr after reperfusion. RESULTS: HTS showed beneficial effects in prevention of liver ischemia/reperfusion injury. HTS groups developed increases in AST and ALT levels that were significantly less than ISS groups; however, the HTS pre-reperfusion group showed levels significantly less than the HTS pre-ischemia group. No differences in IL-6 and IL-10 levels were observed. A significant decrease in mitochondrial dysfunction as well as hepatic edema was observed in the HTS pre-reperfusion group. Pulmonary vascular permeability was significantly less in the pre-reperfusion HTS group compared to the ISS group. No differences in myeloperoxidase activity were observed. The liver histologic score was significantly less in the pre-reperfusion HTS group compared to the pre ischemia HTS group. CONCLUSION: HTS ameliorated local and systemic injuries in experimental liver ischemia/reperfusion. Infusion of HTS in the pre-reperfusion period may be an important adjunct to accomplish the best results. PMID- 20004455 TI - Physiologic predictors of postoperative abscess in children with perforated appendicitis: subset analysis from a prospective randomized trial. AB - BACKGROUND: Intra-abdominal abscess after appendectomy is the most common complication in patients with perforated appendicitis. There are currently little data that may forecast which patients are more likely to develop an abscess. Therefore, we performed a retrospective analysis of a prospectively collected dataset to determine whether there are predictors for developing a postoperative abscess. METHODS: The dataset was collected prospectively in a randomized trial comparing antibiotic regimens in 98 pediatric patients with perforated appendicitis. All patients underwent laparoscopic appendectomy and received a minimum of 5 days of intravenous antibiotics. The Pearson correlation was used to evaluate the influence of patient, intra-operative, and early postoperative variables on the development of an abscess. Two-tailed P values were determined from the correlation coefficient, and significance was defined as P < or = .05. RESULTS: At presentation, a positive correlation for abscess formation was identified with increasing age (P = .003), weight (P = .001), body mass index (P = .008), and diarrhea (P = .005). Operative time had no influence on abscess development. After operation, there was progressively increasing positive correlation between abscess and the maximum temperature each successive postoperative day. This relationship became significant at day 3. An increased white blood cell count on day 5 was highly predictive of abscess (P < .001). CONCLUSION: In children presenting with perforated appendicitis, increasing age, weight, and/or body mass index correlated with the development of a postoperative abscess. Diarrhea on presentation also poses an increased risk of abscess. Postoperatively, each successive day with a fever is incrementally more predictive of an abscess formation. PMID- 20004456 TI - The phosphatase Shp2 is required for signaling by the Kaposi's sarcoma-associated herpesvirus viral GPCR in primary endothelial cells. AB - Kaposi's sarcoma-associated herpesvirus (KSHV) is the causative agent of Kaposi's sarcoma (KS), an AIDS-related endothelial cell malignancy that is the most common cancer in central and southern Africa. The KSHV viral G protein-coupled receptor (vGPCR) is a viral oncogene that conveys a survival advantage to endothelial cells and causes KS-like tumors in mouse models. In this study we investigate the role of Shp2, a protein tyrosine phosphatase in vGPCR signaling. Shp2 is vital to many cytokine-induced signaling pathways and is dysregulated in various infections and malignancies. It has also recently been implicated in angiogenesis. We find that vGPCR activity results in phosphorylation of regulatory tyrosines in Shp2 and that in turn, Shp2 is required for vGPCR mediated activation of MEK, NFkappaB, and AP-1. Furthermore, both genetic and chemical inhibition of Shp2 abrogate vGPCR-induced enhancement of endothelial cell migration. This establishes Shp2 as an important point of convergence of KSHV vGPCR signaling and a potential molecular target in the design of an anti KSHV therapeutic regimen. PMID- 20004457 TI - Induction of angiogenic chemokine CCL2 by human herpesvirus 8 chemokine receptor. AB - Human herpesvirus 8 (HHV-8) is associated with Kaposi's sarcoma (KS), an endothelial cell lesion believed to be initiated and driven primarily by cytokine dysregulation. Among the viral proteins suspected as contributing to viral pathogenesis is the lytically expressed viral G protein-coupled receptor (vGPCR), which can induce various cellular cytokines. CC ligand-2 (CCL2/MCP-1) is a vGPCR regulated angiogenic chemokine found at elevated levels in KS lesions and induced by HHV-8 infection of endothelial cells. Here we show that vGPCR induces CCL2 in endothelial cells via activation of C/EBPbeta and that vGPCR and C/EBPbeta are critical components of CCL2 induction by HHV-8 infection of endothelial cultures. To our knowledge, this is the first report of vGPCR-mediated cytokine induction, and its characterization, in the context of virus infection. Our results identify a mechanism by which vGPCR can contribute, in a host cell shutoff-independent manner, to viral pathogenesis. PMID- 20004459 TI - Quantitative structure activity relationship (QSAR) of chlorine effects on E(LUMO) of disinfection by-product: Chlorinated alkanes. AB - Chlorinated Alkane (CA) is one of the major classes of Disinfection By-Products (DBPs) in chlorinated drinking water. Multi-multivariate regression methods have been used to develop Quantitative Structure-Activity Relationship (QSAR) models between the Energy of the Lowest Unoccupied Molecular Orbital (ELUMO) and three molecular descriptors: namely, number of chlorine (NCl), number of carbon (NC) and the Energy of the Highest Occupied Molecular Orbital (EHOMO). Among different linear QSAR models, the most robust model is ELUMO=-0.1474NCl 0.003766NC+1.9528EHOMO+1.0664 (N=30, R2=0.956, F=188.498, RMSE=0.00762, P=0.000). The model is validated by using internal and external cross validation techniques. The model uncertainties are also quantified through the Bootstrapping and the Taylor methods. Among all the molecular descriptors, number of chlorine (NCl) contributes to the most to the ELUMO of chlorinated alkanes. PMID- 20004458 TI - Ubiquitination of tombusvirus p33 replication protein plays a role in virus replication and binding to the host Vps23p ESCRT protein. AB - Post-translational modifications of viral replication proteins could be widespread phenomena during the replication of plus-stranded RNA viruses. In this article, we identify two lysines in the tombusvirus p33 replication co-factor involved in ubiquitination and show that the same lysines are also important for the p33 to interact with the host Vps23p ESCRT-I factor. We find that the interaction of p33 with Vps23p is also affected by a "late-domain"-like sequence in p33. The combined mutations of the two lysines and the late-domain-like sequences in p33 reduced replication of a replicon RNA of Tomato bushy stunt virus in yeast model host, in plant protoplasts, and plant leaves, suggesting that p33-Vps23p ESCRT protein interaction affects tombusvirus replication. Using ubiquitin-mimicking p33 chimeras, we demonstrate that high level of p33 ubiquitination is inhibitory for TBSV replication. These findings argue that optimal level of p33 ubiquitination plays a regulatory role during tombusvirus infections. PMID- 20004460 TI - Organohalogen contaminants in blood of Kemp's ridley (Lepidochelys kempii) and green sea turtles (Chelonia mydas) from the Gulf of Mexico. AB - The threat that exposure to organohalogen (OH) contaminants poses to endangered populations of Kemp's ridley (Lepidochelys kempii) and green sea turtles (Chelonia mydas) is not well understood, partly because few OH data are available. Blood samples from live juvenile and sub-adult L. kempii (n = 46) and C. mydas (n = 9) from the Gulf of Mexico and from L. kempii from the southeastern US coast (n = 3) were extracted using microwave-assisted extraction, and analyzed by large volume injection gas chromatography-mass spectrometry for 85 polychlorinated biphenyls (PCBs), 25 organochlorine pesticides (OCPs) and 27 polybrominated diphenyl ethers (PBDEs). Plasma chemistries, hematology and immune responses were also assessed. Concentrations of SigmaPCBs (geometric mean, range: 3190 pg g(-1), 227-21590 pg g(-1) blood), SigmaDDTs (geometric mean, range: 541 pg g(-1), 161-4310 pg g(-1) blood) and OCPs in L. kempii from the Gulf were comparable to those reported in L. kempii from the Atlantic. SigmaPBDEs were detected in all samples (geometric mean, range: 146 pg g(-1), 19.5-1450 pg g(-1) blood), with PBDE 47, 99, 100, 153 and 154 being the predominant congeners. SigmaPCBs, SigmaDDTs and Sigmachlordanes were one order of magnitude lower in green turtles, and SigmaPBDE concentrations were lower by half due to trophic level differences. L. kempii from the southeast USA had higher percentages of highly chlorinated PCBs indicating exposure to Aroclor 1268. Blood urea nitrogen was positively correlated to Sigmachlordanes, and SigmaPCBs were inversely correlated to creatine phosphokinase in L. kempii. These data help establish baseline contaminant concentrations in live L. kempii and C. mydas. PMID- 20004461 TI - Determination and assessment of mercury content in calcareous soils. AB - This paper provides the first available information on the determination of the total mercury content in different Mediterranean calcareous soils by thermal decomposition, amalgamation and atomic absorption spectrophotometry. Fifty-three samples from five soil use groups (natural, dry land, greenhouse, irrigated and rice farming soils) were analysed. The results show that the levels of mercury in these soils were 9.4-1585mugkg(-1). Soil organic matter and the zinc equivalent index have been related to Hg content in soils, suggesting that Cu, Ni, Zn and Hg are of a similar origin. The background level (BL), geochemical baseline concentrations (GBC) and the reference value (RV) were established from natural soils. Calculations are according to the log-normal distribution criterion. The BL, GBC and RV values obtained were 25.1, 9.8-64.3, and 64.3mugkg(-1), respectively. The BL was similar to the baseline concentration in soils obtained at Admiralty Bay (Antarctica) (24.6mugkg(-1)) and a Spanish peat bog (Galicia) (22mugkg(-1)). These data show that rice farming soils (gleyic-calcaric Fluvisols, Fluvaquents), irrigated soils (calcaric Fluvisols, Xerofluvents) and some greenhouse soils (cumulic Antrosols) presented much higher levels, indicating contamination. Hg accumulation in these soils was associated with local anthropogenic sources. PMID- 20004462 TI - Atmospheric monitoring of organic pollutants in the Arctic under the Arctic Monitoring and Assessment Programme (AMAP): 1993-2006. AB - Continuous and comparable atmospheric monitoring programs to study the transport and occurrence of persistent organic pollutants (POPs) in the atmosphere of remote regions is essential to better understand the global movement of these chemicals and to evaluate the effectiveness of international control measures. Key results from four main Arctic research stations, Alert (Canada), Pallas (Finland), Storhofdi (Iceland) and Zeppelin (Svalbard/Norway), where long-term monitoring have been carried out since the early 1990s, are summarized. We have also included a discussion of main results from various Arctic satellite stations in Canada, Russia, US (Alaska) and Greenland which have been operational for shorter time periods. Using the Digital Filtration temporal trend development technique, it was found that while some POPs showed more or less consistent declines during the 1990s, this reduction is less apparent in recent years at some sites. In contrast, polybrominated diphenyl ethers (PBDEs) were still found to be increasing by 2005 at Alert with doubling times of 3.5 years in the case of deca-BDE. Levels and patterns of most POPs in Arctic air are also showing spatial variability, which is typically explained by differences in proximity to suspected key source regions and long-range atmospheric transport potentials. Furthermore, increase in worldwide usage of certain pesticides, e.g. chlorothalonil and quintozene, which are contaminated with hexachlorobenzene (HCB), may result in an increase in Arctic air concentration of HCB. The results combined also indicate that both temporal and spatial patterns of POPs in Arctic air may be affected by various processes driven by climate change, such as reduced ice cover, increasing seawater temperatures and an increase in biomass burning in boreal regions as exemplified by the data from the Zeppelin and Alert stations. Further research and continued air monitoring are needed to better understand these processes and its future impact on the Arctic environment. PMID- 20004463 TI - WITHDRAWN: Percutaneous mechanical thrombectomy combined with cathether-directed thrombolysis for lower-extremity deep venous thrombosis. AB - This article has been withdrawn at the request of the author(s) and/or editor. The Publisher apologizes for any inconvenience this may cause. The full Elsevier Policy on Article Withdrawal can be found at http://www.elsevier.com/locate/withdrawalpolicy. PMID- 20004465 TI - DART points the way for HIV treatment programmes. PMID- 20004466 TI - Targeted nonviral delivery vehicles to neural progenitor cells in the mouse subventricular zone. AB - Targeted gene therapy can potentially minimize undesirable off-target toxicity due to specific delivery. Neuron-specific gene delivery in the central nervous system is challenging because neurons are non-dividing and also outnumbered by glial cells. One approach is to transfect dividing neural stem and progenitor cells (NSCs and NPCs, respectively). In this work, we demonstrate cell-specific gene delivery to NPCs in the brains of adult mice using a peptide-modified polymeric vector. Tet1, a 12-amino acid peptide which has been shown to bind specifically to neuronal cells, was utilized as a neuronal targeting ligand. The cationic polymer polyethylenimine (PEI) was covalently modified with polyethylene glycol (PEG) for in vivo salt stability and Tet1 for neuron targeting to yield a Tet1-PEG-PEI conjugate. When plasmid DNA encoding the reporter gene luciferase was complexed with Tet1-PEG-PEI and delivered in vivo via an injection into the lateral ventricle, Tet1-PEG-PEI complexes mediated increased luciferase expression levels in brain tissue when compared to unmodified PEI-PEG complexes. In addition, cells transfected by Tet1-PEG-PEI complexes were found to be exclusively adult NPCs whereas untargeted PEG-PEI complexes were found to transfect a heterogenous population of cells. Thus, we have demonstrated targeted, nonviral delivery of nucleic acids to adult NPCs using the Tet1 targeting ligand. These materials could potentially be used to deliver therapeutic genes for the treatment of neurodegenerative diseases. PMID- 20004467 TI - Induction of DNA double-strand breaks in primary gingival fibroblasts by exposure to dental resin composites. AB - Dental resin composites and their reactive monomers/co-monomers have been shown to elicit cytotoxic responses in human gingival fibroblasts (HGF), and their metabolic radical intermediates have the potential to attack the DNA backbone, which may induce DNA double-strand breaks (DSBs). In this study we have tested the cytotoxicity and induction of DSBs by the most common composite resin monomers/co-monomers: BisGMA, HEMA, TEGDMA, and UDMA in gingival fibroblasts using the sensitive gamma-H2AX DNA repair focus assay. Our results show increasing monomer cytotoxicities in the order of BisGMA>UDMA>TEGDMA>HEMA, an order that was also observed for their capacity to induce DSBs. BisGMA at the EC50 concentration of 0.09 mm evoked the highest rate of gamma-H2AX foci formation that was 11-fold higher DNA DSBs as compared to the negative controls that ranged between 0.25 and 0.5gamma-H2AX foci/HGF cell. Our results for the first time show that exposure to dental resin monomers can induce DSBs in primary human oral cavity cells, which underscores their genotoxic capacity. PMID- 20004468 TI - Protection against titanium particle induced osteolysis by cannabinoid receptor 2 selective antagonist. AB - Osteolysis and subsequent aseptic loosening are the most common causes of failure of total joint arthroplasty. Osteolysis is initiated by inflammatory response to wear debris, resulting in localized, osteoclastic peri-implant bone loss. However, there were no effective measures for prevention and treatment of periprosthetic osteolysis. The aim of the current study was to determine whether CB2 selective antagonist (AM630) inhibits wear debris-induced osteolysis in a murine osteolysis model. Titanium (Ti) particles were introduced into established air pouches on BALB/c mice, followed by implantation of calvaria bone from syngeneic littermates. AM630 was given to mice intraperitoneally 2 days before Ti particles introduction and maintained until the sacrifice of the mice. Mice without drug treatment, as well as mice injected with saline alone, were included. Each group contains 10 mice. Pouch tissues were harvested 14 days after bone implantation for histological and molecular analysis. Ti particles stimulation significantly increased CB2 expression. However, less CB2 was observed in AM630 treatment group. AM630 inhibited Ti particle-induced osteolysis associated gene activity of RANK, RANKL and CPK, and diminished RANKL expression in Ti particle stimulated pouches. AM630 markedly reduced the number of TRAP+ cells in pouch tissues. In conclusion, this study provides the evidence that blockage of CB2 with AM630 can markedly reduce Ti particle induced osteolysis in a murine air pouch model. This finding points to the possibility that CB2 selective antagonists like AM630 may have potential value for prevention and treatment of wear particle induced osteolysis. PMID- 20004464 TI - Routine versus clinically driven laboratory monitoring of HIV antiretroviral therapy in Africa (DART): a randomised non-inferiority trial. AB - BACKGROUND: HIV antiretroviral therapy (ART) is often managed without routine laboratory monitoring in Africa; however, the effect of this approach is unknown. This trial investigated whether routine toxicity and efficacy monitoring of HIV infected patients receiving ART had an important long-term effect on clinical outcomes in Africa. METHODS: In this open, non-inferiority trial in three centres in Uganda and one in Zimbabwe, 3321 symptomatic, ART-naive, HIV-infected adults with CD4 counts less than 200 cells per microL starting ART were randomly assigned to laboratory and clinical monitoring (LCM; n=1659) or clinically driven monitoring (CDM; n=1662) by a computer-generated list. Haematology, biochemistry, and CD4-cell counts were done every 12 weeks. In the LCM group, results were available to clinicians; in the CDM group, results (apart from CD4-cell count) could be requested if clinically indicated and grade 4 toxicities were available. Participants switched to second-line ART after new or recurrent WHO stage 4 events in both groups, or CD4 count less than 100 cells per microL (LCM only). Co primary endpoints were new WHO stage 4 HIV events or death, and serious adverse events. Non-inferiority was defined as the upper 95% confidence limit for the hazard ratio (HR) for new WHO stage 4 events or death being no greater than 1.18. Analyses were by intention to treat. This study is registered, number ISRCTN13968779. FINDINGS: Two participants assigned to CDM and three to LCM were excluded from analyses. 5-year survival was 87% (95% CI 85-88) in the CDM group and 90% (88-91) in the LCM group, and 122 (7%) and 112 (7%) participants, respectively, were lost to follow-up over median 4.9 years' follow-up. 459 (28%) participants receiving CDM versus 356 (21%) LCM had a new WHO stage 4 event or died (6.94 [95% CI 6.33-7.60] vs 5.24 [4.72-5.81] per 100 person-years; absolute difference 1.70 per 100 person-years [0.87-2.54]; HR 1.31 [1.14-1.51]; p=0.0001). Differences in disease progression occurred from the third year on ART, whereas higher rates of switch to second-line treatment occurred in LCM from the second year. 283 (17%) participants receiving CDM versus 260 (16%) LCM had a new serious adverse event (HR 1.12 [0.94-1.32]; p=0.19), with anaemia the most common (76 vs 61 cases). INTERPRETATION: ART can be delivered safely without routine laboratory monitoring for toxic effects, but differences in disease progression suggest a role for monitoring of CD4-cell count from the second year of ART to guide the switch to second-line treatment. FUNDING: UK Medical Research Council, the UK Department for International Development, the Rockefeller Foundation, GlaxoSmithKline, Gilead Sciences, Boehringer-Ingelheim, and Abbott Laboratories. PMID- 20004469 TI - Review: Sex and the human placenta: mediating differential strategies of fetal growth and survival. AB - There are known sex specific differences in fetal and neonatal morbidity and mortality. There are also known differences in birthweight centile with males generally being larger than females at birth. These differences are generally ignored when studying obstetric complications of pregnancy and the mechanisms that confer these differences between the sexes are unknown. Current evidence suggests sex specific adaptation of the placenta may be central to the differences in fetal growth and survival. Our research examining pregnancies complicated by asthma has reported sexually dimorphic differences in fetal growth and survival with males adapting placental function to allow for continued growth in an adverse maternal environment while females reduce growth in an attempt to survive further maternal insults. We have reported sex differences in placental cytokine expression, insulin-like growth factor pathways and the placental response to cortisol in relation to the complication of asthma during pregnancy. More recently we have identified sex specific alterations in placental function in pregnancies complicated by preterm delivery which were associated with neonatal outcome and survival. We propose the sexually dimorphic differences in growth and survival of the fetus are mediated by the sex specific function of the human placenta. This review will present evidence supporting this hypothesis and will argue that to ignore the sex of the placenta is no longer sound scientific practice. PMID- 20004470 TI - Endomorphin-suppressed nitric oxide release from mice peritoneal macrophages. AB - Endomorphins are newly discovered mu-opioid receptor selective immunocompetent opioid peptides. Endomorphin 1 is predominantly distributed in brain, while endomorphin 2 is widely allocated in the spinal cord. Lately, endomorphins have been investigated as modulators of reactive oxygen and nitrogen species. Nitric oxide is short lived radical involved in various biological processes such as regulation of blood vessel contraction, inflammation, neurotransmission and apoptosis. The aim of this work was to investigate the in vivo effects of endomorphins on nitric oxide release and NOS 2 isoenzyme upregulation in mice peritoneal macrophages additionally challenged ex vivo with lipopolysaccharide. The results showed that endomorphin 1 or endomorphin 2 in vitro did not change NO release from peritoneal mouse macrophages during a 48 h incubation period. On the other hand in vivo endomorphins had suppressive effect on NO release as well as on NOS 2 and IL-1 protein concentration. The most of suppressive effect in vivo of both endomorphins was blocked with 30 min pretreatment with mu-receptor selective antagonist beta-FNA, which proved involvement of opioid receptor pathway in suppressive effects of endomorphins. PMID- 20004471 TI - Simultaneous determination of nilotinib, imatinib and its main metabolite (CGP 74588) in human plasma by ultra-violet high performance liquid chromatography. AB - We describe a high performance liquid chromatography (HPLC) method that separates two of the currently licenced tyrosine kinase inhibitors (TKIs); nilotinib (AMN107, Tasigna) and imatinib (STI571, Glivec), together with its main metabolite, CGP-74588, from human plasma. After solid phase extraction the drug mix was separated through a Gemini C6-phenyl column (150 mm x 4.6mm, i.d.; 5 microm) (Phenomenex), UK) under isocratic mobile phase conditions of methanol:50mM ammonium acetate (pH 8) (65:35, v/v) with ultra-violet (UV) detection at 260 nm wavelength. For all compounds the intra-day coefficient of variation and bias were <3% and <5% respectively; and inter-day were <4% and <9%. This simple and novel method may be used to quantify levels of TKIs when used alone or in combination with drug treatments for clinical samples. PMID- 20004472 TI - Pulmonary leukostasis mimicking tuberculosis in endemic area. PMID- 20004473 TI - Early complement factors in the local tissue immunocomplex generated during intestinal ischemia/reperfusion injury. AB - Recent work reveals that the innate immune system is able to recognize self targets and initiate an inflammatory response similar to that of pathogens. One novel example of this innate autoimmunity is ischemia/reperfusion (I/R) injury, in which reperfusion of the ischemic tissues elicits an acute inflammatory response activated by natural IgM (nIgM) binding to ischemia-specific self antigens, which are non-muscle myosin heavy chains type II (NMHC-II) subtype A and C. Subsequently, the complement lectin pathway is activated and eventually tissue injury occurs. Although earlier studies in the intestinal model showed that the classical complement pathway did not initiate I/R injury, C1q deposition was still observed in the local injured tissues by imaging analysis. Moreover, the involvement of the alternative complement pathway became unclear due to conflicting reports using different knockout mice. To explore the immediate downstream pathway following nIgM-ischemic antigen interaction, we isolated the nIgM-ischemic antigen immunocomplexes from the local tissue of animals treated in the intestinal I/R injury model, and examined the presence of initial molecules of three complement pathways. Our results showed that mannan-binding lectin (MBL), the early molecule of the lectin pathway, was present in the nIgM-ischemic Ag immunocomplex. In addition, C1q, the initial molecule of the classical pathway was also detected on the immunocomplex. However, Factor B, the early molecule in the alternative pathway, was not detected in the immunocomplex. To further examine the role of the alternative pathway in I/R injury, we utilized Factor B knockout mice in the intestinal model. Our results showed that Factor B knockout mice were not protected from local tissue injury, and their complement system was activated in the local tissues by nIgM during I/R. These results indicated that the lectin complement pathway operates immediately downstream of the nIgM ischemic antigen interaction during intestinal I/R. Furthermore, the classical complement pathway also appears to interact with the of nIgM-ischemic antigen immunocomplex. Finally, the alternative complement pathway is not involved in I/R injury induction in the current intestinal model. PMID- 20004474 TI - Transglutaminase II interacts with rac1, regulates production of reactive oxygen species, expression of snail, secretion of Th2 cytokines and mediates in vitro and in vivo allergic inflammation. AB - Transglutaminase II (TGase II) is a protein cross-linking enzyme with diverse biological functions. Here we report the role of TGase II in allergic inflammation. Antigen stimulation induced expression and activity of TGase II by activation of NF-kappaB in rat basophilic leukemia (RBL2H3) cells. This induction of TGase II was dependent on FcepsilonRI and EGFR. Interaction between TGase II and rac1 was induced following antigen stimulation. TGase II was responsible for the increased production of reactive oxygen species, expression of prostaglandin E2 synthase (PGE2 synthase) and was responsible for increased secretion of prostaglandin E2. ChIP assay showed that TGase II, through interaction with NF kappaB, was responsible for the induction of histone deacetylase-3 (HDAC3) and snail by direct binding to promoter sequences. HDAC3 and snail induced by TGase II, exerted transcriptional repression on E-cadherin. Snail exerted negative effect on expression of MMP-2, and secretion of Th2 cytokines. Inhibition of matrix metalloproteinase-2 (MMP-2) inhibited secretion of Th2 cytokines. In vivo induction of TGase II was observed in Balb/c mouse model of IgE antibody-induced passive cutaneous anaphylaxis. Chemical inhibition of TGase II exerted negative effect on IgE-dependent passive cutaneous anaphylaxis. Chemical inhibition of TGase II by cystamine exerted negative effect on Balb/c mouse model of phorbol myristate acetate (PMA)-induced atopic dermatitis. These results suggest novel role of TGase II in allergic inflammation and TGase II can be developed as target for the development of allergy therapeutics. PMID- 20004475 TI - Humoral immunity in tuberculin skin test anergy and its role in high-risk persons exposed to active tuberculosis. AB - The most common test to identify latent tuberculosis is the tuberculin skin test that detects T cell responses of delayed type hypersensitivity type IV. Since it produces false negative reactions in active tuberculosis or in high-risk persons exposed to tuberculosis patients as shown in this report, we studied antibody profiles to explain the anergy of such responses in high-risk individuals without active infection. Our results showed that humoral immunity against tuberculin, regardless of the result of the tuberculin skin test is important for protection from active tuberculosis and that the presence of high antibody titers is a more reliable indicator of infection latency suggesting that latency can be based on the levels of antibodies together with in vitro proliferation of peripheral blood mononuclear cells in the presence of the purified protein derivative. Importantly, anti-tuberculin IgG antibody levels mediate the anergy described herein, which could also prevent reactivation of disease in high-risk individuals with high antibody titers. Such anti-tuberculin IgG antibodies were also found associated with blocking and/or stimulation of in vitro cultures of PBMC with tuberculin. In this regard, future studies need to establish if immune responses to Mycobacterium tuberculosis can generate a broad spectrum of reactions either toward Th1 responses favoring stimulation by cytokines or by antibodies and those toward diminished responses by Th2 cytokines or blocking by antibodies; possibly involving mechanisms of antibody dependent protection from Mtb by different subclasses of IgG. PMID- 20004476 TI - An evaluation of an integrated model of relapse in depression. AB - BACKGROUND AND AIM: The present study evaluated an integrated model of the role of psychosocial factors in the prediction of relapse of Major Depressive Disorder over a one-year follow-up period. METHODS AND ANALYSES: A range of established variables, including life stress, cognitive-personality vulnerability factors, social support, and coping responses, were simultaneously considered in a series of prediction models, in an adult female sample of remitted depressed individuals. RESULTS: It was determined that interpersonal marked difficulties, social support, and emotion-oriented and avoidance-oriented coping provided the best-fitting and most parsimonious predictive model for depressive relapse at one year follow-up. CONCLUSIONS: The examination of multifactorial models of risk represents a promising avenue for future research and theory development. PMID- 20004477 TI - The relationship of demographic, clinical, cognitive and personality variables to the discrepancy between self and clinician rated depression. AB - BACKGROUND: The measurement of depression severity is an important aspect of both clinical and research practices. However, studies examining the self-report Beck Depression Inventory (BDI) and the clinician Hamilton Depression Rating Scale indicate only moderate correlations. The aim of this study was to examine the correlation between two self-report measures, the revised BDI, the Hopkins Symptom Checklist with the clinician rated, Montgomery-Asberg Depression Rating Scale. The secondary aim was to investigate patient factors which contribute to discordant ratings. METHODS: Depression severity and demographic, clinical, personality, cognitive, and personality factors that may contribute to a self report-clinician rated discrepancy were examined in 177 adult outpatients with a Major Depressive Episode (DSM-IV) participating in a randomised clinical trial comparing CBT and IPT for depression. All assessment was conducted prior to treatment randomisation. RESULTS: Self-report and clinician rated depression were moderately correlated. Individuals with higher clinician rated depression severity, increased levels of rumination and females were more likely to have higher self-report rated depression (BDI-II and SCL-90) than clinician rated depression. In addition, younger patients and those with melancholic depression had higher BDI-II compared to MADRS scores. LIMITATIONS: Results require replication. CONCLUSIONS: Self-reported and observer rated depression were only moderately correlated. Researchers and clinicians interpreting the level of depression need to be cognizant of the patient factors that may contribute to either underreporting or overreporting self-report scores relative to observer ratings. PMID- 20004478 TI - Genetic risk, parent-child relations, and antisocial phenotypes in a sample of African-American males. AB - Gene x environment interactions have been found to be associated with the development of antisocial behaviors. The extant gene x environment research, however, has failed to measure directly the ways in which global measures of genetic risk may interact with a putative environmental risk factor. The current study addresses this gap in the literature and examines the interrelationships among a global measure of genetic risk based on five genetic polymorphisms, a measure of parent-child relations, and eight antisocial phenotypes. Analysis of African-American males (N = 145 to 159) drawn from the National Longitudinal Study of Adolescent Health (Add Health) revealed two broad findings. First, the genetic risk and parent-child relations scales were inconsistently related to the outcome variables. Second, genetic risk and parent-child relations interacted to predict variation in all of the eight antisocial phenotype measures. These findings point to the possibility that measures of genetic risk that are based on multiple polymorphisms can be employed to examine the gene x environmental basis to antisocial behavioral phenotypes. PMID- 20004479 TI - Dissociation of decisions in ambiguous and risky situations in obsessive compulsive disorder. AB - Patients with obsessive-compulsive disorder (OCD) often show deficits in everyday decision-making, a phenomenon which is leading to a growing research interest in neuropsychological aspects of decision-making in OCD. Previous investigations of OCD patients demonstrated deficits in the Iowa Gambling Task (IGT), a decision making task with implicit rules. Results were interpreted as reflecting orbitofrontal cortex dysfunctions observed in OCD. The aim of the present study is to investigate OCD patients' performance on the Game of Dice Task (GDT), a decision-making task with explicit and stable rules. For this purpose, 23 patients with OCD and 22 healthy comparison subjects were examined with the GDT and the IGT as well as with tests of executive functioning. While patients performed worse than comparison subjects on the IGT, they were unimpaired on the GDT and executive functioning tasks. Results further emphasize dysfunctions of the orbitofrontal cortex, but indicate intact functioning of the dorsolateral prefrontal cortex in patients with OCD. PMID- 20004480 TI - Catechol-O-methyltransferase Val158Met polymorphism affects therapeutic response to mood stabilizer in symptomatic manic patients. AB - Catechol-O-methyltransferase (COMT) is a candidate gene for the pathogenesis of some psychiatric disorders. The aim of this study is to examine the role of the COMT gene Val158Met polymorphism on the clinical aspects of bipolar disorder including symptomatology and therapeutic response. This study comprised 144 unrelated manic patients who met strict DSM-IV criteria for bipolar I disorder and 157 healthy unrelated controls. All subjects were of Korean ethnicity. To evaluate the clinical symptoms, we used the Young Mania Rating Scale at baseline and 6 weeks after treatment. No statistically significant difference in genotype distribution was found between manic patients and normal controls. There was also no significant difference in symptomatology among the genotypes in manic patients. In therapeutic response, however, patients with the Met/Met genotype were significantly more frequent in the non-responder than in the responder group. Our results suggest that the COMT gene polymorphism in the therapeutic response to mood stabilizers in manic patients. Further studies with a larger number of subjects and well-controlled design will be required to better understand the role of the COMT gene polymorphism on the therapeutic response to mood stabilizer in manic patients. PMID- 20004481 TI - Impulse-control disorders in children and adolescents with obsessive-compulsive disorder. AB - The aim of this study was to examine current prevalences, clinical correlates and patterns of co-occurrence of impulse-control disorders (ICDs) in children and adolescents with obsessive-compulsive disorder (OCD). We examined rates and clinical correlates of comorbid ICDs in 70 consecutive child and adolescent subjects with lifetime DSM-IV OCD (32.9% females; mean age = 13.8 +/- 2.9 years). Comorbidity data were obtained with structured clinical interviews using DSM-IV criteria. OCD severity was assessed with the Child Yale-Brown Obsessive Compulsive Scale. All variables were compared in OCD subjects with and without current ICDs. 12 (17.1%) subjects met criteria for a current ICD. Pathological skin picking and compulsive nail biting were the most common ICDs with current rates of 12.8% and 10.0%, respectively. OCD subjects with current ICDs were significantly more likely to have a co-occurring tic disorder (66.7% vs. 20.7%). Although having an ICD was associated with greater numerical scores of OCD symptomatology, these differences were not statistically significant. There were no sex-specific patterns of ICD occurrence in children and adolescents with OCD. Certain ICDs are common among children and adolescents with OCD. Better identification of ICDs in children and adolescents with OCD is needed, as are empirically validated treatments for youth with co-occurring ICDs. PMID- 20004482 TI - Changes in hypothalamic-pituitary-thyroid axis following successful treatment with low-frequency right prefrontal transcranial magnetic stimulation in treatment-resistant depression. AB - Hypothalamic-pituitary-thyroid (HPT) axis abnormalities have been reported in some patients with major depression. To knowledge, however, the effects of low frequency right prefrontal transcranial magnetic stimulation (TMS) on the HPT axis have not yet been elucidated. The goal of this study was to evaluate alterations in the HPT axis associated with the therapeutic efficacy of TMS treatments. Twenty patients with treatment-resistant depression received five 60 s 1-Hz trains over the right dorsolateral prefrontal cortex. Twelve treatment sessions were administered within a 3-week period (total pulses, 3600). Responders were defined as a > or =50% decrease in the Hamilton Depression Rating Scale (HDRS) score. Serum levels of thyroid-stimulating hormone (TSH), free triiodothyronine (fT3), and free thyroxine (fT4) were measured, respectively, at pre- and post-treatment. There were no significant changes in fT3 and fT4 levels measured at either pre- or post-treatment in either responders or nonresponders; however, TSH levels of responders elevated significantly after TMS treatments. In addition, there was a significant negative correlation between TSH levels at pretreatment and decrease (%) in the HDRS score. These findings suggest that the HPT axis is associated with antidepressant effects of low-frequency right prefrontal TMS, and indicate that lower TSH levels at pre-treatment are correlated with better therapeutic efficacy. PMID- 20004484 TI - Molecular structural and functional characterization of STAT1 gene regulatory region in teleost Channa argus. AB - The transcription factor STAT1 is involved in signal transduction of type I and II interferons (IFNs). However, the molecular characteristics of the STAT1 regulatory region still remain to be elucidated in teleosts. In the present study, the complete cDNA and the regulatory region of the STAT1 gene were isolated from snakehead (Channa argus). More than 2.4kb 5'-flanking region of STAT1 shares the regulatory elements of IFN-stimulated response element (ISRE) and IFN-gamma activation site (GAS). Consensus ISRE and GAS were located from 373 to -361 and -716 to -724 in the promoter region, respectively. Moreover, it is noticeable that the crucial elements of ISRE (+698 to +710) and GAS (+294 and +301) are present in the first intron of snakehead STAT1. Comparisons of six vertebrate STAT1 5'-flanking regions all present the common sequence characteristics of IFN-induced gene promoter, which include ISRE, GAS and Sp1 sites. In order to further characterize the snakehead STAT1 regulatory region, six reporter constructs of snakehead STAT1 promoter and first intron were generated to examine the specificity to human interferon-gamma (hIFN-gamma). Only those constructs containing the ISRE element showed notable reporter activity after stimulation of Hela cells with hIFN-gamma. However, sequential deletions of putative transcription factor binding sites indicated that GAS elements have little effect on the promoter and intronic activity in response to hIFN-gamma. Taken together, these results suggest that the regulatory mechanisms of IFN signalling appear to be mediated in a similar manner in fish and mammals. PMID- 20004483 TI - Feline leukemia virus immunity induced by whole inactivated virus vaccination. AB - A fraction of cats exposed to feline leukemia virus (FeLV) effectively contain virus and resist persistent antigenemia/viremia. Using real-time PCR (qPCR) to quantitate circulating viral DNA levels, previously we detected persistent FeLV DNA in blood cells of non-antigenemic cats considered to have resisted FeLV challenge. In addition, previously we used RNA qPCR to quantitate circulating viral RNA levels and determined that the vast majority of viral DNA is transcriptionally active, even in the absence of antigenemia. A single comparison of all USDA-licensed commercially available FeLV vaccines using these modern sensitive methods has not been reported. To determine whether FeLV vaccination would prevent nucleic acid persistence, we assayed circulating viral DNA, RNA, antigen, infectious virus, and virus neutralizing (VN) antibody in vaccinated and unvaccinated cats challenged with infectious FeLV. We identified challenged vaccinates with undetectable antigenemia and viremia concomitant with persistent FeLV DNA and/or RNA. Moreover, these studies demonstrated that two whole inactivated virus (WIV) adjuvanted FeLV vaccines (Fort Dodge Animal Health's Fel O-Vax Lv-K) and Schering-Plough Animal Health's FEVAXYN FeLV) provided effective protection against FeLV challenge. In nearly every recipient of these vaccines, neither viral DNA, RNA, antigen, nor infectious virus could be detected in blood after FeLV challenge. Interestingly, this effective viral containment occurred despite a weak to undetectable VN antibody response. The above findings reinforce the precept of FeLV infection as a unique model of effective retroviral immunity elicited by WIV vaccination, and as such holds valuable insights into retroviral immunoprevention and therapy. PMID- 20004485 TI - The complexities of antiretroviral drug-drug interactions: role of ABC and SLC transporters. AB - Treatment of human immunodeficiency virus (HIV) infection involves a combination of several antiviral agents belonging to different pharmacological classes. This combination is referred to as highly active antiretroviral therapy (HAART). This treatment has proved to be very effective in suppressing HIV replication, but antiretroviral drugs have complex pharmacokinetic properties involving extensive drug metabolism and transport by membrane-associated drug carriers. Combination drug therapy often introduces complex drug-drug interactions that can result in toxic or sub-therapeutic drug concentrations, compromising treatment. This review focuses on the role of ATP-binding cassette (ABC) membrane-associated efflux transporters and solute carrier (SLC) uptake transporters in antiretroviral drug disposition, and identifies clinically important antiretroviral drug-drug interactions associated with changes in drug transport. PMID- 20004486 TI - Biomarker responses in polar cod (Boreogadus saida) exposed to the water soluble fraction of crude oil. AB - In order to mimic the biological effects of an oil spill in Arctic waters, we examined several types of biomarkers (genes, enzymes, metabolites, and DNA damage) in polar cod Boreogadus saida experimentally exposed to the water soluble fractions of crude oil. During 4 weeks of exposure, induction of the studied biomarkers exceeded baseline levels. The mRNA expression of the cytochrome P4501A1 (cyp1a1) gene was the most promising biomarker, with glutathione S transferase (gst) as a suitable complement. The delayed ethoxyresorufin O deethylase (EROD) and GST activities and their persistence following 2 weeks of depuration may allow detection of previous exposures in field samples. The composition of PAH metabolites in the bile indicated the bioavailability of different PAH size-classes. Although mRNA expressions of antioxidant defense genes were induced at start of the exposure, with the strongest responses from catalase and cytosolic superoxide dismutase, they were poor for oil monitoring purposes due to their very short response times. Significant DNA damage demonstrated genotoxicity even at low PAH concentrations (<15microgL(-1)) and was correlated with benzo(a)pyrene and pyrene metabolites in the bile. PMID- 20004487 TI - Landscape risk factors for attacks of vampire bats on cattle in Sao Paulo, Brazil. AB - Vampire-bat (Desmodus rotundus) attacks on cattle are a major concern for cattle raising area. Blood loss and paralytic rabies due to bat bites can impose severe losses on the livestock. We took four municipalities inside the Sao Joao da Boa Vista veterinary district (Sao Paulo, Brazil) as a study area and tested a set of landscape features for spatial correlation with distance to areas in which vampire-bat attacks on cattle were documented. Bat- and cattle-related data from the Sao Paulo State Rabies Control Program were used. Landscape data (first-order rivers and their tributaries, main roads, railways and urban areas) were obtained from official cartographic agencies; forest, sugarcane and pasture data were acquired from remote-sensing mappings. The study area was taken as a grid split into 178 cells. Each 4kmx4km cell was filled with bat, cattle and landscape data. Our analysis detected that grid cells that were closer to areas of bat attacks on cattle had higher cattle density and a greater percentage of the land committed to sugarcane cropping, and were close to forest fragments. These results shed light on the need for rethink the Rabies Control Program strategies for defining the surveillance of vampire-bat populations and rabies control in the state of Sao Paulo, Brazil. PMID- 20004488 TI - Does graded return-to-work improve sick-listed workers' chance of returning to regular working hours? AB - Using Danish register and survey data, we examine the effect of a national graded return-to-work program on the probability of sick-listed workers returning to regular working hours. During program participation, the sick-listed worker works fewer hours and receives the normal hourly wage for the hours worked and sickness benefit for the hours off work. When the worker's health improves, working hours are increased until the sick-listed worker is able to work regular hours. Taking account of unobserved differences between program participants and non participants, we find that participation in the program significantly increases the probability of returning to regular working hours. PMID- 20004489 TI - Hospital closure and economic efficiency. AB - We present a new framework for assessing the effects of hospital closures on social welfare and the local economy. While patient welfare necessarily declines when patients lose access to a hospital, closures also tend to reduce costs. We study five hospital closures in two states and find that urban hospital bailouts reduce aggregate social welfare: on balance, the cost savings from closures more than offset the reduction in patient welfare. However, because some of the cost savings are shared nationally, total surplus in the local community may decline following a hospital closure. PMID- 20004490 TI - Evaluating the policy of setting up microscopy centres at township hospitals in Shandong China: experience from patients and providers. AB - BACKGROUND: Since 2004 the China Ministry of Health policy has required microscopy centres (MCs) to be set up in one third of township hospitals nationally, to improve the accessibility of sputum smear testing for TB in rural areas. The objective of this study was to assess the performance of MCs in Shandong province from both patient and provider perspectives. METHODS: A survey of 245 TB suspects was conducted in 8 counties of Shandong stratified by MC performance. Seventy-two health providers and administrators were interviewed at the township and county levels. RESULTS: General performance of MC was poor. In 2006, the high and low performance groups checked on average 190 and 24 TB suspects, respectively. The majority of TB suspects who visited a MC did not have their sputum checked, or sputum was checked but the result was not recorded. TB suspects who visited a MC tended to live closer to it and had better knowledge of the MC than those who visited the county TB dispensary (CTD) directly. Patients with severe TB symptoms tended to go directly to the CTD. No significant difference in medical expenses before the TB diagnosis or diagnostic delay was found between TB suspects who visited a MC and those who did not. Several reasons were identified. The policy tried to set up too many MCs regardless of transportation conditions. It lacked operational details. Township hospitals had limited funding, qualified staff, and technical support from the CTD. The existing referral incentive discouraged sputum checks at the MC. CONCLUSION: The national MC policy fell short of its goals in Shandong. Neither patients nor providers were interested in using MC in its current form. Policy recommendations are given. PMID- 20004491 TI - Costs of hospital-based methadone maintenance treatment in HIV/AIDS control among injecting drug users in Indonesia. AB - OBJECTIVE: To assess the cost of hospital-based methadone maintenance treatment (MMT) for injecting drug users (IDUs) in Bandung, Indonesia; to address concerns of financial sustainability at the hospital level and financial accessibility and economic attractiveness at the health care policy level. METHODS: In a 1 year observation period in 2006-2007, MMT service delivery costs were estimated on the basis of a micro-costing approach. Patient costs were estimated on the basis of a survey among 48 methadone clients. RESULTS: A total number of 129 clients attended the MMT clinic, resulting in a total of 16,335 client visits. Total annual societal costs of running the MMT clinic equalled Rp 1130 mln (US$123,672), or Rp 69,206 (US$7.57) per client visit. Of total costs, patient costs established the largest share (65%), followed by that of central government (20%), and the hospital (15%). Present consultation tariffs already cover hospital costs and the patient costs of accessing MMT services constitute almost 70% of their income. CONCLUSION: Under current circumstances, MMT services are financially sustainable to the hospital. MMT services are subsidized by the central government, and this is warranted considering the important role of the program in HIV/AIDS among IDUs. Still, the present user fee seems a barrier to utilization, and a higher level of subsidy might be justified to reduce the cost to the patient. PMID- 20004492 TI - An intensity-region driven multi-classifier scheme for improving the classification accuracy of proteomic MS-spectra. AB - In this study, a pattern recognition system is presented for improving the classification accuracy of MS-spectra by means of gathering information from different MS-spectra intensity regions using a majority vote ensemble combination. The method starts by automatically breaking down all MS-spectra into common intensity regions. Subsequently, the most informative features (m/z values), which might constitute potential significant biomarkers, are extracted from each common intensity region over all the MS-spectra and, finally, normal from ovarian cancer MS-spectra are discriminated using a multi-classifier scheme, with members the Support Vector Machine, the Probabilistic Neural Network and the k-Nearest Neighbour classifiers. Clinical material was obtained from the publicly available ovarian proteomic dataset (8-7-02). To ensure robust and reliable estimates, the proposed pattern recognition system was evaluated using an external cross-validation process. The average overall performance of the system in discriminating normal from cancer ovarian MS-spectra was 97.18% with 98.52% mean sensitivity and 94.84% mean specificity values. PMID- 20004493 TI - Parallel computation of mutual information on the GPU with application to real time registration of 3D medical images. AB - Due to processing constraints, automatic image-based registration of medical images has been largely used as a pre-operative tool. We propose a novel method named sort and count for efficient parallelization of mutual information (MI) computation designed for massively multi-processing architectures. Combined with a parallel transformation implementation and an improved optimization algorithm, our method achieves real-time (less than 1s) rigid registration of 3D medical images using a commodity graphics processing unit (GPU). This represents a more than 50-fold improvement over a standard implementation on a CPU. Real-time registration opens new possibilities for development of improved and interactive intraoperative tools that can be used for enhanced visualization and navigation during an intervention. PMID- 20004494 TI - Planning forwards: biodiversity research and monitoring systems for better management. PMID- 20004495 TI - SIRT2-mediated protein deacetylation: An emerging key regulator in brain physiology and pathology. AB - Protein function is considerably altered by posttranslational modification. In recent years, cycles of acetylation/deacetylation emerged as fundamental regulators adjusting biological activity of many proteins. Particularly, protein deacetylation by Sirtuins, a family of atypical histone deacetylases (HDACs), was demonstrated to regulate fundamental cell biological processes including gene expression, genome stability, mitosis, nutrient metabolism, aging, mitochondrial function and cell motility. Given this wealth of biological functions, perhaps not unexpectedly then, pharmacological compounds targeting Sirtuin activity are now prime therapeutic agents for alleviating severity of major diseases encompassing diabetes, cancer, cardiovascular and neurodegenerative disorders in many organs. In this review, we will focus on the brain and its physiological and pathological processes governed by Sirtuin-mediated deacetylation. Besides discussing Sirtuin function in neurodegenerative diseases, emphasis will be given on the mounting evidence deciphering key developmental brain functions for Sirtuins in neuronal motility, neuroprotection and oligodendrocyte differentiation. In this respect, we will particularly highlight functions of the unconventional family member SIRT2 in post-mitotic neurons and glial cells. PMID- 20004496 TI - The complex regulation of WRKY53 during leaf senescence of Arabidopsis thaliana. AB - Many different agriculturally important traits, e.g. number and quality of seeds, timing of seed set, fruit ripening, are affected by senescence. Despite the importance of the senescence processes in plants, our knowledge on regulatory mechanisms of senescence is still poor. A central step is a massive reprogramming of the transcriptome, implying an important role of transcription factors. In Arabidopsis 12-16% of all genes are up- or down-regulated. WRKY transcription factors play a central role in controlling leaf senescence in Arabidopsis. One important member of this family, WRKY53, is tightly regulated by different unexpected mechanisms and is a convergence node between senescence and biotic and abiotic stress responses. PMID- 20004497 TI - Changes in chloroplast ultrastructure, fatty acid components of thylakoid membrane and chlorophyll a fluorescence transient in flag leaves of a super-high yield hybrid rice and its parents during the reproductive stage. AB - In plants, it is well established that chloroplast is one of the early targgeted organelles to breakdown during leaves senescing. Here we applied a newly developed super-high-yield hybrid rice (Oryza sativa) LiangYouPeiJiu (LYPJ) and its parents lines to investigate changes in ultrastructure of chloroplasts, fatty acid composition of thylakoid membrane lipids and chlorophyll (Chl) a fluorescence transient in natural senescing leaves. We found that at full expansion of flag leaves in three lines, chloroplasts often showed oblong shapes with a typical membrane system of stroma and grana thylakoids, whereas their shapes had been changed from oblong to spherical during senescence. Our data showed that the initiation of senescence displayed accumulation of starch and an increase in the number and size of plastoglobuli with the damaged thylakoid membranes; subsequently, swollen thylakoid membranes in stroma and in grana with a significant increase in MDA content, and disorganization of thylakoid membrane system with significant changes in fatty acid composition of thylakoid membrane lipids were developed. Compared with its parents, the newly developed hybrid rice LYPJ had the highest capacity of carbohydrate transport from leaves (sources) to ears (sink), marked with the lowest accumulation of starch grains in the leaf chloroplasts, and the slowest senescing rate of chloroplast in overall leaf senescence process. Chl a fluorescence transients of three kinds of flag leaves were analyzed by so-called JIP-test. The results of analysis suggest that these findings inculding a high inherited activity of antioxidant enzymes and high photosynthetate transport to pretect chloroplast structure in the hybrid rice LYPJ have close relations to its super-high yield. PMID- 20004498 TI - Early detection of age-related memory deficits in individual mice. AB - To date, no consensus has been reached concerning the age of the earliest onset of age-related cognitive deficits in rodents. Our aim was to develop a behavioral model allowing early and individual detection of age-related cognitive impairments. We tested young (3 months), middle-aged (10 months) and aged (17 months) C57Bl/6 mice in the starmaze, a task allowing precise analysis of the search pattern of mice via standardized calculation of two navigation indices. We performed mouse-per-mouse analyses and compared each mouse's performance to a threshold based on young mice's performances. Using this method we identified impaired mice from the age of 10 months old. Their deficits were independent of any sensorimotor dysfunctions and were associated with an alteration of the maintenance of the hippocampal CA1 late-LTP. This study develops reliable methodology for early detection of age-related memory disorders and provides evidence that memory can decline in some individuals as early as from the age of 10 months. PMID- 20004499 TI - A randomized controlled trial of estrogen treatment in men with mild cognitive impairment. AB - This randomized, placebo-controlled, cross-over study investigated whether estrogen treatment would have a beneficial effect on tests of verbal memory in men with mild cognitive impairment (MCI). Forty-three men newly diagnosed with MCI were administered a battery of neuropsychological tests before randomly receiving 12 weeks of treatment with estrogen or placebo followed by a 12 week cross-over treatment. A significant improvement in the total score, and in two subscale scores of the Buschke Selective Reminding Test occurred following estrogen treatment compared to both pretreatment and post-placebo scores (p<0.05). However, benefit occurred only in the men who had received estrogen for 12 weeks following 12 weeks of placebo. Although these findings tentatively suggest that treatment with estrogen may improve verbal memory in men with MCI, the fact that the improvement occurred only in the group that received estrogen following 12 weeks of placebo and the absence of improvement on every test of verbal memory administered suggests that these findings need to be replicated using a larger sample size. PMID- 20004500 TI - Use of multitracers for the study of water mixing in the Paraiba do Sul River estuary. AB - Multitracers were used to study water mixing in the Paraiba do Sul River estuary region in August 2007 (dry season) and March 2008 (rainy season) and to evaluate the reach of the river plume in the direction of the open ocean. Two sampling campaigns were carried out, each in a different season. Based on these results, it was possible to conclude that the multitracers used in this study (salinity, Si, Ba and U, as well as the radium isotopes 223Ra, 224Ra, 226Ra and 228Ra) presented satisfactory results toward defining the plume reach and determining the residence time and water-mixing processes in the estuary. A strong correlation was observed between tracers and the distance to the coast. During the low river water discharge period, the riverine water took about 10 days to reach open ocean waters (salinity approximately 35). During the rainy period this value decreased to 6 days. Based on the radium results, it was possible to calculate diffusion coefficients (K(h)) of 23 km(2) d(-1) and 38 km(2) d(-1) for 224Ra and 223Ra, respectively, during the dry season (winter). Values of 65 km(2) d(-1) and 68 km(2) d(-1) for 223Ra and 224Ra, respectively, were found for the rainy period (summer). PMID- 20004501 TI - Gait parameters and stride-to-stride variability during familiarization to walking on a split-belt treadmill. AB - BACKGROUND: Subjects unfamiliar to walking on a split-belt treadmill may initially demonstrate an altered gait pattern or increased variability of gait parameters. While previous investigations have examined kinematic variables associated with familiarization time, the objective of this study was to determine the familiarization period required to obtain the most reproducible gait pattern through the assessment of kinetic, kinematic and spatio-temporal parameters during a single session of treadmill walking. METHODS: Eleven healthy subjects participated in a single bout of treadmill walking which lasted 9 min. Kinematic and kinetic data were collected from the first 30s of each minute, beginning when the treadmill reached full speed. Means and standard deviations for knee flexion at heel strike, ground reaction forces, step width and step length were obtained to examine the changes in each variable over the 9 min. Mean r(2) values were evaluated for changes in variability from one stride to the subsequent stride for sagittal plane hip, knee and ankle joint angles and moments, as well as for vertical and horizontal ground reaction forces. FINDINGS: Significant reductions in variability were found for vertical and horizontal ground reaction forces, knee flexion at heel strike and step length over 9 min. Only step width showed a change in the mean value across trials. There were no increases in r(2) values after the 5th min for any of the gait variables. INTERPRETATION: The results suggest that in order to collect accurate data for gait analysis, subjects should be familiarized to the split-belt treadmill for at least 5 min prior to data collection. PMID- 20004502 TI - Cruciate ligament tensile forces during the forward and side lunge. AB - BACKGROUND: Although weight bearing lunge exercises are frequently employed during anterior cruciate ligament and posterior cruciate ligament rehabilitation, cruciate ligament tensile forces are currently unknown while performing forward and side lunge exercises with and without a stride. METHODS: Eighteen subjects used their 12 repetition maximum weight while performing a forward lunge and side lunge with and without a stride. A motion analysis system and biomechanical model were used to estimate cruciate ligament forces during lunging as a function of 0 90 degrees knee angles. FINDINGS: Comparing the forward lunge to the side lunge across stride variations, mean posterior cruciate ligament forces ranged between 205 and 765N and were significantly greater (P<0.0025) in the forward lunge long at 40 degrees , 50 degrees , 60 degrees , 70 degrees , and 80 degrees knee angles of the descent phase and at 80 degrees , 70 degrees , 60 degrees knee angles of the ascent phase. There were no significant differences (P<0.0025) in mean posterior cruciate ligament forces between with and without stride differences across lunging variations. There were no anterior cruciate ligament forces quantified while performing forward and side lunge exercises. INTERPRETATION: Clinicians should be cautious in prescribing forward and side lunge exercises during early phases of posterior cruciate ligament rehabilitation due to relatively high posterior cruciate ligament forces that are generated, especially during the forward lunge at knee angles between 40 degrees and 90 degrees knee angles. Both the forward and side lunges appear appropriate during all phases of anterior cruciate ligament rehabilitation. Understanding how forward and side lunging affect cruciate ligament loading over varying knee angles may help clinicians better prescribe lunging exercises in a safe manner during anterior cruciate ligament and posterior cruciate ligament rehabilitation. PMID- 20004503 TI - Comparison of trunk stiffness provided by different design characteristics of lumbosacral orthoses. AB - BACKGROUND: Lumbosacral orthoses (LSOs) are class I medical devices that are used in conservative and postoperative management of low back pain. The effectiveness of LSOs depends on their design aimed at enhancing trunk stiffness. Therefore, the purpose of this study was to compare two lumbar supports: extensible (made of neoprene and lycra) and non-extensible (made of polyester and nylon). METHODS: Trunk stiffness and damping was estimated from trunk displacement data in response to a quick force release in trunk flexion, extension, and lateral bending. Fourteen male and 6 female subjects performed five trials at each experimental condition: (1) No LSO, (2) extensible LSO, (3) non-extensible LSO, (4) non-extensible LSO with a small rigid front panel, and (5) non-extensible LSO with a large rigid front panel. Testing order was randomized and the LSOs were cinched to a pressure of 70 mmHg (9.4 kPa) measured between posterior aspect of the iliac crest and the orthosis. FINDINGS: The non-extensible LSO reduced trunk displacement by 14% and increased trunk stiffness by 14% (P<0.001). The extensible LSO did not result in any significant change in trunk displacement or stiffness. The addition of rigid front panels to the non-extensible LSO did not improve its effectiveness. The trunk damping did not differ between the LSO conditions. INTERPRETATION: A non-extensible LSO is more effective in augmenting trunk stiffness and limiting trunk motion following a perturbation than an extensible LSO. The rigid front panels do not provide any additional trunk stiffness most likely due to incongruence created between the body and a brace. PMID- 20004504 TI - Teres major muscle activation relates to clinical outcome in tendon transfer surgery. AB - BACKGROUND: In massive rotator cuff tears a teres major (TMj) tendon transfer to the insertion of the supraspinatus (SSp) reverses its adduction moment arm into abduction which is supposed to be an adequate salvage procedure. Analysis of muscle function to find biomechanical ground of such success is scarce. METHODS: We compared pre- and postoperative clinical outcome of TMj transfer, i.e. Range of Motion, pain, Constant Shoulder scores and arm force. TMj activation was evaluated in 14 patients suffering massive cuff tears using activation ratios to describe the desired 'in-phase' and undesired 'out-of-phase' contribution to the external arm moment. Additionally, we analyzed activation of the latissimus dorsi (LD) and the medial part of the deltoids (DE). The activation ratios were compared to controls and TMj activation ratios were related to clinical outcome. FINDINGS: TMj tendon transfer improved arm function. Pre-operatively, we observed 'out-of-phase'abduction activation of TMj and LD. After transfer patients activated TMj according to its new anatomical position. 'Out-of-phase' LD abduction activation persisted. The clinical improvements coincided with changes in activation ratio of TMj. INTERPRETATION: 'Out-of-phase' TMj adductor activation is associated with compromised arm function in patients with irreparable cuff tears. After transfer, TMj is activated in correspondence with its new anatomical function, which was supportive for the improved arm function. PMID- 20004505 TI - Widespread occurrence of estrogenic UV-filters in aquatic ecosystems in Switzerland. AB - We performed a trace analytical study covering nine hormonally active UV-filters by LC-MS/MS and GC-MS in river water and biota. Water was analysed at 10 sites above and below wastewater treatment plants in the river Glatt using polar organic chemical integrative samplers (POCIS). Four UV-filters occurred in the following order of decreasing concentrations; benzophenone-4 (BP-4) > benzophenone-3 (BP-3) > 3-(4-methyl)benzylidene-camphor (4-MBC) > 2-ethyl-hexyl-4 trimethoxycinnamate (EHMC). BP-4 ranged from 0.27 to 24.0 microg/POCIS, BP-3, 4 MBC and EHMC up to 0.1 microg/POCIS. Wastewater was the most important source. Levels decreased with higher river water flow. No significant in-stream removal occurred. BP-3, 4-MBC and EHMC were between 6 and 68 ng/L in river water. EHMC was accumulated in biota. In all 48 macroinvertebrate and fish samples from six rivers lipid-weighted EHMC occurred up to 337 ng/g, and up to 701 ng/g in 5 cormorants, suggesting food-chain accumulation. UV-filters are found to be ubiquitous in aquatic systems. PMID- 20004506 TI - [Impact of videothoracoscopic sympathectomy in thoracic surgery. Future considerations]. PMID- 20004507 TI - [Tracheobronchomalacia]. AB - Tracheobronchomalacia is a central airway disease characterised by weakness of the wall and dynamic decrease in the tracheal lumen and the large bronchi, particularly while exhaling. It is more common in middle age and the elderly with previous exposure to cigarettes. It causes chronic symptoms such as cough, dyspnea, increase in recurrent infections, and poor secretion management, but it can also progress to chronic respiratory failure and death. It is usually confused with other common diseases like chronic obstructive pulmonary disease (COPD) or asthma. Its causes can be congenital or acquired and its diagnosis involves the dynamic assessment of the airway with tomography and fibrobronchoscopy. It is classified as mild, moderate or severe depending on the degree of collapse of the airway when exhaling. Management consists of a primary phase, in which concomitant diseases must be controlled, such as COPD, asthma or gastro-oesophageal reflux. In diffuse moderate to severe symptomatic tracheobronchomalacia tracheobronchoplasty must be considered with strengthening of the posterior wall. Silicone and "Y" stents can be used to identify patients who could potentially benefit from surgical treatment as well as being used for the definitive symptomatic treatment with high surgical risk. More prospective studies need to be done in order to standardise certain common criteria for the management of this usually under-diagnosed disease. PMID- 20004508 TI - Microvascular flow during mechanical cardiopulmonary resuscitation. PMID- 20004509 TI - Mix proportions and properties of CLSC made from thin film transition liquid crystal display optical waste glass. AB - In this study, controlled low-strength concrete (CLSC) is mixed using different water-to-binder (W/B) ratios (1.1, 1.3 and 1.5) and various percentages of sand substituted by waste LCD glass sand (0%, 10%, 20% and 30%). The properties of the fresh concrete, including compressive strength, electrical resistivity, ultrasonic pulse velocity, permeability ratio and shrinking of the CLSC, are examined. Results show that increases in amount of waste glass added result in better slump and slump flow, longer initial setting time and smaller unit weight. Compressive strength decreases with increasing W/B ratio and greater amounts of waste glass added. Both electrical resistivity and ultrasonic pulse velocity increase with increases in amount of waste glass and decreases in W/B ratio. On the contrary, the permeability ratio increases with increases in W/B ratio, but decreases with greater amounts of waste glass added. CLSC specimens cured for different durations show little changes in length with shrinkage below 0.025%. Our findings reveal that CLSC mixed using waste LCD glass in place of sand can meet design requirements. Recycling of waste LCD glass not only offers an economical substitute for aggregates, but also an ecological alternative for waste management. PMID- 20004511 TI - Epidemiological and clinical features of Moyamoya disease in Nanjing, China. AB - OBJECTIVE: The epidemiology of Moyamoya disease in mainland China has not been documented. Therefore, the present study was designed to examine the epidemiological and clinical features of Moyamoya disease in Nanjing, a provincial capital in China. METHODS: Patient records from multiple hospitals in Nanjing from January 2000 to December 2007 were collected. The clinical features of Moyamoya disease were retrospectively analyzed. RESULTS: A total of 202 patients were identified. There were 94 males and 108 females, with ages ranging from 2 to 78 years. There was a dual age peak, one in the group of patients 5-9 years of age and another in the group of patients 35-39 years of age. The initial symptoms included cerebral ischemia (81 patients, 40%), cerebral hemorrhage (113 patients, 55.9%) and asymptomatic disease (8 patients, 3.9%). An increasing incidence rate of Moyamoya disease was observed during the period of 2000-2007, with an average detection rate of 0.43cases/100,000 persons/year (prevalence 3.92/100,000 persons). The incidence of ischemia associated with the disease was 0.16cases/100,000 people-years and the incidence of hemorrhage was 0.22cases/100,000 people-years. CONCLUSION: This first study on the epidemiological and clinical features of Moyamoya disease in mainland China indicated an increasing incidence of Moyamoya disease with bimodal incidence distribution appearing more frequently in adults. PMID- 20004513 TI - Predicting competitive adsorption behavior of major toxic anionic elements onto activated alumina: a speciation-based approach. AB - Toxic anionic elements such as arsenic, selenium, and vanadium often co-exist in groundwater. These elements may impact each other when adsorption methods are used to remove them. In this study, we investigated the competitive adsorption behavior of As(V), Se(IV), and V(V) onto activated alumina under different pH and surface loading conditions. Results indicated that these anionic elements interfered with each other during adsorption. A speciation-based model was developed to quantify the competitive adsorption behavior of these elements. This model could predict the adsorption data well over the pH range of 1.5-12 for various surface loading conditions, using the same set of adsorption constants obtained from single-sorbate systems. This model has great implications in accurately predicting the field capacity of activated alumina under various local water quality conditions when multiple competitive anionic elements are present. PMID- 20004514 TI - Study of the filtration performance of a plain wave fabric filter using response surface methodology. AB - The gas-solid two-phase flows in the plain wave fabric filter were simulated by computational fluid dynamics (CFD) technology, and the warps and wefts of the fabric filter were made of filaments with different dimensions. The numerical solutions were carried out using commercial computational fluid dynamics (CFD) code Fluent 6.1. The filtration performances of the plain wave fabric filter with different geometry parameters and operating condition, including the horizontal distance, the vertical distance and the face velocity were calculated. The effects of geometry parameters and operating condition on filtration efficiency and pressure drop were studied using response surface methodology (RSM) by means of the statistical software (Minitab V14), and two second-order polynomial models were obtained with regard to the effect of the three factors as stated above. Moreover, the models were modified by dismissing the insignificant terms. The results show that the horizontal distance, vertical distance and the face velocity all play an important role in influencing the filtration efficiency and pressure drop of the plane wave fabric filters. The horizontal distance of 3.8 times the fiber diameter, the vertical distance of 4.0 times the fiber diameter and Reynolds number of 0.98 are found to be the optimal conditions to achieve the highest filtration efficiency at the same face velocity, while maintaining an acceptable pressure drop. PMID- 20004512 TI - MicroRNA in pancreatic cancer: pathological, diagnostic and therapeutic implications. AB - MicroRNAs (miRNAs) are a group of small non-coding RNA molecules of 17-25 nucleotides (nt) in length, predicted to control the activity of about 30% of all protein-coding genes in mammals. Altered expressions of miRNAs are reported in various cancers and may associate with cancer pathogenesis, apoptosis, and cell growth, thereby functioning as either tumor suppressors or oncogenes. Recent reports showed that deregulation of miRNA contribute to tumor development and progression and hence, have diagnostic and prognostic value in several human malignancies. This review discusses the current status of miRNA in pancreatic cancer development, progression, diagnosis, and therapy. PMID- 20004515 TI - Enzymatic mechanism and biochemistry for cyanide degradation: a review. AB - Cyanides are fast-acting poisons, can be lethal if exposed in excess. In spite of fact, cyanides are discharged as effluents in large scale from industries every year. Certain bacteria, fungi, algae and plants produce cyanides. It has been observed that microbes and plant systems can degrade cyanides to less toxic compounds. There are many enzymes, which are produced by microorganisms that utilize cyanides as substrate to make alanine, glutamic acid, alfa-amino-butyric acid, beta-cyanoalanine, etc. Present paper deals with different enzymes, their mechanisms and corresponding pathways with respect to the known biochemistry of enzyme and feasibility for the use in treatment of cyanides containing industrial effluents. PMID- 20004516 TI - Reduction in toxicity of arsenic(III) to Halobacillus sp. Y35 by kaolin and their related adsorption studies. AB - The growth of Halobacillus sp. Y35 has been investigated in HGM hypersaline medium with different doses of As(III) and kaolin. The metabolic heat flux decreases with the increase in As(III) concentration, indicating that strain Y35 lowers their metabolic activity in order to resist the As(III) toxicity. Carbon dioxide flux, cell growth and protein synthesis rates, and total thermal effect have been, for the first time, successfully employed simultaneously to assess the effect of As(III) on strain Y35 in the absence and presence of kaolin. The relative adsorption capacity and adsorption intensity of kaolin for As(III) are higher with strain Y35 than that without strain Y35, demonstrating that it is possible to reduce the toxicity of As(III) to our environment by both using mineral adsorption and biosorption technology. Our work shows the potential application of kaolin and strain Y35 for the removal of As(III) from contaminated groundwater. PMID- 20004517 TI - Hydrothermal-hydrolysis synthesis and photocatalytic properties of nano-TiO2 with an adjustable crystalline structure. AB - Tri-phase (anatase, rutile, and brookite), bi-phase (anatase and rutile), and mono-phase (rutile) TiO(2) nanomaterials with different morphologies were successively synthesized using a hydrothermal-hydrolysis method and adjusting the Ti(4+)/Ti(3+) molar ratio in a precursor solution. The properties of the fabricated nanomaterials were characterized using X-ray diffraction (XRD), scanning electron microscopy (SEM), transmission electron microscopy (TEM), photocatalytic reaction, and other techniques. It has been shown that TiO(2) nanorods can be obtained by increasing the Ti(4+)/Ti(3+) molar ratio in a precursor solution from 1:0 to 0.3:0.7. TiO(2) nanoparticles are formed if the Ti(3+) fraction in the solution is further increased. The selective synthesis of TiO(2) nanomaterials is explained by a decrease in the reaction rate and by changes in acidity with increasing Ti(3+) content. The tri-phase nanorods and bi phase nanoparticles synthesized with Ti(4+)/Ti(3+) molar ratios from 1:0 to 0.8:0.2 and 0.2:0.8 to 0:1, respectively, have a higher degradation ability with respect to methylene blue aqueous solutions under UV irradiation at ambient temperature compared to purely rutile TiO(2) nanorods synthesized with Ti(4+)/Ti(3+) molar ratios from 0.7:0.3 to 0.3:0.7. The high photocatalytic activity of the multi-phase TiO(2) samples is primarily attributed to their larger band gap and suppressed recombination of photo-generated electron-hole pairs. PMID- 20004518 TI - Thermodynamic modeling of ferric phosphate precipitation for phosphorus removal and recovery from wastewater. AB - Phosphorus removal and recovery by ferric phosphate (FePO(4) x 2 H(2)O) precipitation has been considered as an effective technology. In the present study, we examined chemical precipitation thermodynamic modeling of the PHREEQC program for phosphorus removal and recovery from wastewater. The objective of this research was to employ thermodynamic modeling to evaluate the effect of solution factors on FePO(4) x 2 H(2)O precipitation. In order to provide comparison, with the evaluation of thermodynamic modeling, the case study of phosphate removal from anaerobic supernatant was studied. The results indicated that the saturation-index (SI) of FePO(4) x 2 H(2)O followed a polynomial function of pH, and the solution pH influenced the ion activities of ferric iron salts and phosphate. The SI of FePO(4) x 2 H(2)O increased with a logarithmic function of Fe(3+):PO(4)(3-) molar ratio (Fe/P) and initial PO(4)(3-) concentration, respectively. Furthermore, the SI of FePO(4) x 2 H(2)O decreased with a logarithmic function of alkalinity and ionic strength, respectively. With an increase in temperature, the SI at pH 6.0 and 9.0 decreased with a linear function, and the SI at pH 4.0 followed a polynomial function. For the case study of phosphate removal from anaerobic supernatant, the phosphate removal trend at different pH and Fe/P was closer to the predictions of thermodynamic modeling. The results indicated that the thermodynamic modeling of FePO(4) x 2 H(2)O precipitation could be utilized to predict the technology parameters for phosphorus removal and recovery. PMID- 20004519 TI - Degradation of 4-nitrophenol (4-NP) using Fe-TiO2 as a heterogeneous photo-Fenton catalyst. AB - Photocatalytic degradation of 4-nitrophenol was investigated using Fe-doped (1, 3, 5 and 8 wt.% Fe) TiO(2) catalysts under UV light irradiation in aqueous dispersions in the presence of H(2)O(2). Photocatalysts with the lowest Fe content (1%) showed a considerably better behavior with respect to the unloaded TiO(2) and the catalysts with higher Fe contents. Photocatalytic degradation was studied under different conditions such as amounts of 1% Fe-TiO(2) catalyst, H(2)O(2) dose and initial pH of 4-NP solution. The results indicated that about 67.53% total organic carbon of a solution containing 20 mg L(-1) 4-NP was removed at pH 6.17 by using 4.9 mM of H(2)O(2) and 0.4 g L(-1) of the catalyst in a 2-L batch photo-reactor, the complete degradation of 4-NP occurring after 60 min. It was also observed that catalytic behavior could be reproduced in consecutive experiments without a considerable decrease of the UV/Fe-TiO(2)/H(2)O(2) process efficiency. PMID- 20004520 TI - Separation and preconcentration of trace level of lead in one drop of blood sample by using graphite furnace atomic absorption spectrometry. AB - Drop-to-drop solvent microextraction (DDSME) assisted with ultrasonication is applied for the determination of lead in one drop (30 microL) of blood sample by using graphite furnace atomic absorption spectrometry (GF-AAS). The optimum extraction efficiency of lead was observed for 10 min extraction time at pH 5.0 with 2 microL of organic solvent that containing 0.5 M of Cyanex-302. The optimized methodology exhibited good linearity in the range of 0.3-30.0 ng mL(-1) lead with relative standard deviations (RSD) from 2.5 to 4.4%. The method is found to be simple and rapid for the analysis of lead in micro amount of blood sample with the limit of detection (LOD) of 0.08 ng mL(-1). The application of the proposed method has been successfully tested for the determination of lead in blood samples. The results showed that under the optimized experimental conditions, the method showed good sensitivity and recovery %, as well as advantages such as linearity, simplicity, low cost and high feasibility. PMID- 20004522 TI - Extracts of Calpurnia aurea leaves from southern Ethiopia attract and immobilise or kill ticks. AB - Calpurnia aurea extracts are used in southern Ethiopia to protect stock against ticks. Acetone, hexane and water leaf extracts of C. aurea collected in southern Ethiopia were tested for repellent/attractant and acaricidal properties on unfed adult Rhipicephalus pulchellus ticks. In contrast to many other plant species evaluated, C. aurea extracts did not have repellent properties, but rather had a slight attractant capacity. With 20% and 10% acetone extracts, all ticks were either killed or their mobility severely compromised after 1microl of extract was topically applied on the abdomen. At a 5% concentration, 85% of ticks were still affected. A 10% aqueous solution also had a marked effect. The results prove the efficacy of the traditional use of this extract and may lead to a product that can be used commercially to protect animals against tick infestation, under subsistence as well as industrialized conditions. PMID- 20004524 TI - Asymmetry of protracted tongue: can it replace the Wada test? AB - Some asymmetry of body parts (thumbs, cubital crease levels, popliteal crease levels, toes, breasts, gonads and facial structures) is commonly observed. And there is also neurobehavioral asymmetry such as handedness and language lateralization. Since right-handedness is closely associated with left-hemisphere dominance for language, it is known that handedness can be linked with language lateralization. But because cultural and social pressures influence handedness, handedness is able to shift. If so, handedness can't always reflect language lateralization. We can also detect another asymmetry in protracted tongue which is thought to be inherently unchangeable. Some midline deviation of fully protracted tongue is observed in every person. Protractor of the tongue, genioglossus muscle, is solely innervated by contralateral corticobulbar fibers from the inferior part of precentral gyrus. This part is closely related with language expressive area. Therefore, the midline deviation of tongue observed when it is fully protracted could be more related with language lateralization than handedness changeable to external pressures. PMID- 20004525 TI - A phase I/II trial of gefitinib given concurrently with radiotherapy in patients with nonmetastatic prostate cancer. AB - PURPOSE: To estimate the safety and tolerability of daily administration of 250 mg of gefitinib given concurrently with three-dimensional conformal radiotherapy for patients with nonmetastatic prostate cancer. METHODS AND MATERIALS: A total of 42 patients with T2-T3N0M0 tumors were treated in a nonrandomized single center study. A prostate-specific antigen (PSA) level of <20 and a good performance status (WHO, 0-1) were required. Adjuvant or neoadjuvant hormone treatments were not allowed. A daily regimen of 250 mg of gefitinib was started 1 week before radiation therapy began and lasted for the duration of radiation therapy. A dose of 50.4 Gy (1.8 Gy/day) was administered to the tumor, prostate, and seminal vesicles, followed by a 22-Gy booster (2 Gy/day) for a total dose of 72.4 Gy. Correlative studies included analysis of epidermal growth factor receptor (EGFR), EGFRvIII, and phosphorylated EGFR in tumors and tumor necrosis factor, interleukin-1alpha (IL-1alpha), and IL-6 in serum. RESULTS: Maximum tolerated dose was not reached in phase I (12 patients), and 30 additional patients were treated in phase II. Thirty (71.4%) patients completed trial medication. Dose-limiting toxicities were recorded for 16 (38.1%) patients, the most common of which was a grade 3 to 4 increase in transaminase (6 patients). After a median follow-up of 38 months, there were no deaths due to prostate cancer. The estimated PSA relapse-free survival rate at 4 years (Kaplan-Meier) was 97%, the salvage therapy-free survival rate was 91%, and the overall survival rate was 87%. These figures compared favorably with those of matched patients treated with radiation only at higher doses. CONCLUSIONS: The combination of gefitinib and radiation is reasonably well tolerated and has promising activity against nonmetastatic prostate cancer. PMID- 20004526 TI - Relating dose outside the prostate with freedom from failure in the Dutch trial 68 Gy vs. 78 Gy. AB - PURPOSE: For prostate cancer patients at risk for subclinical spread of the disease, we investigated whether incidental dose outside the target was associated with tumor control. METHODS AND MATERIALS: We selected 352 intermediate-risk (mainly T2b-T3a) and high-risk (mainly T3b) patients treated in a randomized trial. Target volume was prostate (68-78 Gy) and seminal vesicles (50-78 Gy). Failure (clinical or biochemical) was evaluated at 4 years. To compare three-dimensional dose distributions, an automated mapping procedure was introduced. Between patients, these maps provide an approximate identification of corresponding anatomical locations. Maps of the dose difference between patients with and without failure were constructed. Univariate and multivariate analyses were performed including the dose in selected points. RESULTS: Dose differences were mainly found in the obturatorial region for the high-risk patients, and in the presacral region for the intermediate risk group (>7 Gy, p < 0.01). Univariate hazard ratios per 10 Gy for selected dose points were 0.83 (p = 0.01, obturatorial) and 0.72 (p = 0.002, presacral). These hazard ratios were stable under multivariate analysis correcting for established prognostic factors, hospital, and dose to the prostate. CONCLUSIONS: Patients without failure have received on average a higher dose to regions where regional cancer spread could be expected. PMID- 20004523 TI - Community-dwelling cocaine-dependent men and women respond differently to social stressors versus cocaine cues. AB - There are likely to be gender differences in determinants of relapse to drug use following abstinence in cocaine-dependent individuals. Cocaine-dependent women are more likely to attribute relapse to negative emotional states and interpersonal conflict. Cocaine dependence has also been linked to dysregulation of stress response and the hypothalamic pituitary adrenal (HPA) axis which may differ between genders. Subjective and HPA-axis responses to a social evaluative stressor, the Trier Social Stress Test (TRIER), and in vivo cocaine-related cues were examined in the present study. RESULTS: There were no gender differences in magnitude of craving responses to the TRIER or the CUE. Both genders had a greater craving response to the CUE than to the TRIER, but the magnitude of the difference was greater for men than women (p=0.04). Cocaine-dependent subjects, compared to the control group, had significantly higher response throughout the TRIER (p<0.0001) and CUE (p<0.0001) testing sessions. There were no gender differences and no gender by cocaine interaction for ACTH responses to the TRIER, although women had lower baseline ACTH (p=0.049). On the CUE task, in contrast, female cocaine-dependent subjects had a more blunted ACTH response than did the other three groups (p=0.02). Female cocaine-dependent subjects also had a lower odds of a positive cortisol response to the TRIER as compared to the other three groups (OR=0.84, 95% CI=[0.02, 1.01]). During the CUE task, cocaine-dependent subjects had overall higher mean cortisol levels (p=0.0001), and higher odds of demonstrating a positive cortisol response to the CUE (OR=2.61, 95% CI=[1.11, 6.11]). No gender differences were found in ACTH responses to the CUE. The results are reviewed in the context of the existing literature on gender differences in cocaine dependence and potential implications for treatment are discussed. PMID- 20004527 TI - Three-dimensional conformal radiation therapy for esophageal squamous cell carcinoma: is elective nodal irradiation necessary? AB - PURPOSE: To evaluate the local control, survival, and toxicity associated with three-dimensional conformal radiotherapy (3D-CRT) for squamous cell carcinoma (SCC) of the esophagus, to determine the appropriate target volumes, and to determine whether elective nodal irradiation is necessary in these patients. METHODS AND MATERIALS: A prospective study of 3D-CRT was undertaken in patients with esophageal SCC without distant metastases. Patients received 68.4 Gy in 41 fractions over 44 days using late-course accelerated hyperfractionated 3D-CRT. Only the primary tumor and positive lymph nodes were irradiated. Isolated out-of field regional nodal recurrence was defined as a recurrence in an initially uninvolved regional lymph node. RESULTS: All 53 patients who made up the study population tolerated the irradiation well. No acute or late Grade 4 or 5 toxicity was observed. The median survival time was 30 months (95% confidence interval, 17.7-41.8). The overall survival rate at 1, 2, and 3 years was 77%, 56%, and 41%, respectively. The local control rate at 1, 2, and 3 years was 83%, 74%, and 62%, respectively. Thirty-nine of the 53 patients (74%) showed treatment failure. Seventeen of the 39 (44%) developed an in-field recurrence, 18 (46%) distant metastasis with or without regional failure, and 3 (8%) an isolated out-of-field nodal recurrence only. One patient died of disease in an unknown location. CONCLUSIONS: In patients treated with 3D-CRT for esophageal SCC, the omission of elective nodal irradiation was not associated with a significant amount of failure in lymph node regions not included in the planning target volume. Local failure and distant metastases remained the predominant problems. PMID- 20004528 TI - Reduction of dose delivered to organs at risk in prostate cancer patients via image-guided radiation therapy. AB - PURPOSE: To determine whether image guidance can improve the dose delivered to target organs and organs at risk (OARs) for prostate cancer patients treated with intensity-modulated radiotherapy (IMRT). METHODS AND MATERIALS: Eight prostate cancer patients were treated with IMRT to 76 Gy at 2 Gy per fraction. Daily target localization was performed via alignment of three intraprostatic fiducials and weekly kV-cone beam computed tomography (CBCT) scans. The prostate and OARs were manually contoured on each CBCT by a single physician. Daily patient setup shifts were obtained by comparing alignment of skin tattoos with the treatment position based on fiducials. Treatment fields were retrospectively applied to CBCT scans. The dose distributions were calculated using actual treatment plans (an 8-mm PTV margin everywhere except for 6-mm posteriorly) with and without image guidance shifts. Furthermore, the feasibility of margin reduction was evaluated by reducing planning margins to 4 mm everywhere except for 3 mm posteriorly. RESULTS: For the eight treatment plans on the 56 CBCT scans, the average doses to 98% of the prostate (D98) were 102% (range, 99-104%) and 99% (range, 45-104%) with and without image guidance, respectively. Using margin reduction, the average D98s were 100% (range, 84-104%) and 92% (range, 40-104%) with and without image guidance, respectively. CONCLUSIONS: Currently, margins used in IMRT plans are adequate to deliver a dose to the prostate with conventional patient positioning using skin tattoos or bony anatomy. The use of image guidance may facilitate significant reduction of planning margins. Future studies to assess the efficacy of decreasing margins and improvement of treatment related toxicities are warranted. PMID- 20004529 TI - Tomotherapy and multifield intensity-modulated radiotherapy planning reduce cardiac doses in left-sided breast cancer patients with unfavorable cardiac anatomy. AB - PURPOSE: For patients with left-sided breast cancers, radiation treatment to the intact breast results in high doses to significant volumes of the heart, increasing the risk of cardiac morbidity, particularly in women with unfavorable cardiac anatomy. We compare helical tomotherapy (TOMO) and inverse planned intensity modulated radiation therapy (IMRT) with three-dimensional conformal radiotherapy using opposed tangents (3D-CRT) for reductions in cardiac volumes receiving high doses. METHODS AND MATERIALS: Fifteen patients with left-sided breast cancers and unfavorable cardiac anatomy, determined by a maximum heart depth (MHD) of >or=1.0 cm within the tangent fields, were planned for TOMO and IMRT with five to seven beam angles, in addition to 3D-CRT. The volumes of heart and left ventricle receiving >or=35 Gy (V35) were compared for the plans, as were the mean doses to the contralateral breast and the volume receiving >or=20 Gy (V20) for the ipsilateral lung. RESULTS: The mean MHD was 1.7 cm, and a significant correlation was observed between MHD and both heart and left ventricle V35. The V35s for IMRT (0.7%) and TOMO (0.5%) were significantly lower than for 3D-CRT (3.6%). The V20 for IMRT (22%) was significantly higher than for 3D-CRT (15%) or TOMO (18%), but the contralateral breast mean dose for TOMO (2.48 Gy) was significantly higher than for 3D-CRT (0.93 Gy) or IMRT (1.38 Gy). CONCLUSIONS: Both TOMO and IMRT can significantly reduce cardiac doses, with modest increases in dose to other tissues in left-sided breast cancer patients with unfavorable cardiac anatomy. PMID- 20004530 TI - Comparison of planned dose distributions calculated by Monte Carlo and Ray-Trace algorithms for the treatment of lung tumors with cyberknife: a preliminary study in 33 patients. AB - PURPOSE: To compare dose distributions calculated using the Monte Carlo algorithm (MC) and Ray-Trace algorithm (effective path length method, EPL) for CyberKnife treatments of lung tumors. MATERIALS AND METHODS: An acceptable treatment plan is created using Multiplan 2.1 and MC dose calculation. Dose is prescribed to the isodose line encompassing 95% of the planning target volume (PTV) and this is the plan clinically delivered. For comparison, the Ray-Trace algorithm with heterogeneity correction (EPL) is used to recalculate the dose distribution for this plan using the same beams, beam directions, and monitor units (MUs). RESULTS: The maximum doses calculated by the EPL to target PTV are uniformly larger than the MC plans by up to a factor of 1.63. Up to a factor of four larger maximum dose differences are observed for the critical structures in the chest. More beams traversing larger distances through low density lung are associated with larger differences, consistent with the fact that the EPL overestimates doses in low-density structures and this effect is more pronounced as collimator size decreases. CONCLUSIONS: We establish that changing the treatment plan calculation algorithm from EPL to MC can produce large differences in target and critical organs' dose coverage. The observed discrepancies are larger for plans using smaller collimator sizes and have strong dependency on the anatomical relationship of target-critical structures. PMID- 20004531 TI - The megavoltage radiation therapy in treatment of patients with advanced or difficult giant cell tumors of bone. AB - PURPOSE: To assess the outcomes of radiotherapy, in terms of local control and treatment complications, of advanced or difficult giant cell tumors of bone (GCTB) that could not be treated by surgery. METHODS AND MATERIALS: Among 122 consecutive patients with confirmed GCTB from 1985 to 2007, 77 patients were treated by megavoltage radiotherapy because they were inappropriate candidates for surgery. We have performed analysis of all data in terms of progression-free survival (PFS) and treatment morbidity. Median follow-up time was 58 months. RESULTS: In the irradiated group, maximal tumor size ranged from 5 to 18 cm (median, 8.5). Anatomic distribution was as follows: femur, 27 cases; tibia, 19; radial/ulnar bone, 12; sacrum, 9; pelvic bones, 5; other, 5. Twenty-one patients (27%) were referred for local recurrence after >=1 other treatment procedures. The radiation doses ranged from 26 to 89 Gy (median, 56; administered 1.8-2.0 Gy/fraction with average total duration of treatment of 5-7 weeks); 8 patients (10%) received <50 Gy. All patients tolerated treatment well without acute or late complications. All patients except two are alive. Local control was achieved in 65 patients (84%; bone recalcification/restitution of joint functions), 12 patients showed signs of local progression, all within irradiated fields (9 were treated successfully with salvage surgery). Five- and 10-year local PFS were 83% and 73%, respectively. Three patients developed lungs metastases. Malignant transformation of GCTB occurred in two patients. CONCLUSIONS: GCTB can be safely and effectively treated with megavoltage radiotherapy with local control rate >80% at 5 years. Our study confirms that radiotherapy of GCTB offers an alternative to difficult or complex surgery and may be an option of choice in the treatment of inoperable patients. PMID- 20004532 TI - Tumor volume reduction rate measured by magnetic resonance volumetry correlated with pathologic tumor response of preoperative chemoradiotherapy for rectal cancer. AB - PURPOSE: To determine whether the tumor volume reduction rate (TVRR) measured using three-dimensional region-of-interest magnetic resonance volumetry correlates with the pathologic tumor response after preoperative chemoradiotherapy (CRT) for locally advanced rectal cancer. METHODS AND MATERIALS: The study included 405 patients with locally advanced rectal cancer (cT3-T4) who had undergone preoperative CRT and radical proctectomy. The tumor volume was measured using three-dimensional region-of-interest magnetic resonance volumetry before and after CRT but before surgery. We analyzed the correlation between the TVRR and the pathologic tumor response in terms of downstaging and tumor regression grade (TRG). Downstaging was defined as ypStage 0-I (ypT0 T2N0M0), and the TRG proposed by Dworak et al. was used. RESULTS: The mean TVRR was 65.0% +/- 22.3%. Downstaging and complete regression occurred in 167 (41.2%) and 58 (14.3%) patients, respectively. The TVRRs according to ypT classification (ypT0-T2 vs. ypT3-T4), ypN classification (ypN0 vs. ypN1-N2), downstaging (ypStage 0-I vs. ypStage II-III), good regression (TRG 3-4 vs. TRG 1-2), and complete regression (TRG 4 vs. TRG 1-3) were all significantly different (p <.05). When the TVRR was categorized into three groups (<60%, 60-80%, and >80%), the rates of ypT0-T2, ypN0, downstaging, and good regression were all significantly greater for patients with a TVRR of >or=60%, as was the complete regression rate for patients with a TVRR >80% (p <.05). CONCLUSION: The TVRR measured using three-dimensional region-of-interest magnetic resonance volumetry correlated significantly with the pathologic tumor response in terms of downstaging and TRG after preoperative CRT for locally advanced rectal cancer. PMID- 20004533 TI - Clinical applicability of cone-beam computed tomography in monitoring seroma volume change during breast irradiation. AB - PURPOSE: To determine whether cone-beam CT (CBCT) is effective in monitoring seroma reduction during breast irradiation when compared with conventional CT. PATIENTS AND METHODS: This study included 19 women with Stage T1-2 breast cancer treated with breast-conserving therapy. Each patient underwent two to four CT and multiple CBCT scans (mean, 8; range, 7-13 scans) at various time intervals during radiotherapy. Seroma were contoured by two observers on all scans and checked by one radiation oncologist. Seroma clarity was determined according to The British Columbia Cancer Agency Seroma Clarity Score scale, and conformity index (CI) of the two observers was evaluated. Correlations in seroma contours and seroma characteristics between CBCT and CT, as well as interobserver variation, were examined. RESULTS: The mean differences in seroma volume between CT and CBCT (3%, p = 0.3) and between the two observers (6%, p = 0.2) were not statistically significant. Seroma clarity correlated significantly with CI for both CT and CBCT (p = 0.02 and p = 0.001, respectively), indicating the higher the seroma clarity score, the greater the CI between the observers. With seroma clarity 3 or higher for CT and CBCT, a high level of observer concordance was shown (all CI of these scans were >or=50%). CONCLUSION: Volume discrepancy between CBCT and CT and between the two observers was not statistically significant. Seroma clarity influenced observers' ability to contour on CT or CBCT equally. Therefore, CBCT is a good clinical surrogate for CT in monitoring seroma reduction during breast radiotherapy, especially for patients with seroma clarity score 3 or higher. PMID- 20004534 TI - The impact of radiotherapy fields in the treatment of patients with choroid plexus carcinoma. AB - PURPOSE: To perform a comprehensive literature review and analysis of cases dealing with choroid plexus carcinoma (CPC) to determine the optimal radiotherapy (RT) treatment field. METHODS AND MATERIALS: A PubMed search of English language articles from 1979 to 2008 was performed, yielding 33 articles with 56 patients who had available data regarding RT treatment field. The median age at diagnosis was 2.7 years (range, 1 month-53 years). Of 54 patients with data regarding type of surgery, 21 (38.9%) had complete resection. Chemotherapy was delivered to 27 (48%) as part of initial therapy. The RT treatment volume was the craniospinal axis in 38 (68%), whole brain in 9 (16%), and tumor/tumor bed in 9 (16%). Median follow-up for surviving patients was 40 months. RESULTS: The 5-year overall survival and progression-free survival (PFS) rates were 59.5% and 37.2%, respectively. Complete resection (p = 0.035) and use of craniospinal irradiation (CSI; p = 0.025) were found to positively affect PFS. The 5-year PFS for patients who had CSI vs. whole brain and tumor/tumor bed RT were 44.2% and 15.3%. For the 19 patients who relapsed, 9 (47%) had a recurrence in the RT field, 6 (32%) had a recurrence outside the RT field, and 4 (21%) had a recurrence inside and outside the irradiated field. CONCLUSION: Patients with CPC who received CSI had better PFS compared with those receiving less than CSI. This study supports the use of CSI in the multimodality management of patients with CPC. PMID- 20004536 TI - Reduced cerebellar diameter in very preterm infants with abnormal general movements. AB - BACKGROUND: Abnormal General Movements (GMs) early in life are predictive of later neuromotor deficits and are related to white matter abnormalities on magnetic resonance imaging (MRI). However, other structural correlates of abnormal GMs have not been defined. AIMS: The objective of this study was to explore brain-metrics (linear brain measurements on MRI representative of 3-D brain volumes) at term as a predictor of abnormal GMs at 1 and 3months' corrected age in preterm infants. It was hypothesized that abnormal GMs would be related to reduced brain-metrics in primary motor areas, namely the cerebellum and parietal lobes. STUDY DESIGN: Eighty three preterm infants (<30weeks' gestational age) were scanned at term-equivalent age. MRI was assessed for white matter abnormality and brain-metrics in six predefined brain regions (i.e. bifrontal, biparietal, lateral ventricles and transverse cerebellar diameters, and inter hemispheric distance). OUTCOME MEASURES: At 1 and 3months' corrected age infants' GMs were assessed from video-taped footage and rated as normal or abnormal using standardized methodology. RESULTS: At 1month, 63% (n=52) of infants had abnormal GMs with no association between any of the brain-metrics and abnormal GMs. At 3months, 23% (n=18) of infants had abnormal GMs (absent fidgety movements n=18; abnormal fidgety movements n=0). Reduced bifrontal, biparietal, and cerebellar transverse diameters, along with an increase in lateral ventricle sizes were associated with an increased risk of abnormal GMs at 3months' corrected age. After controlling for white matter abnormality and grade III/IV intraventricular haemorrhage, only the cerebellar transverse diameter was predictive of abnormal GMs at 3months. CONCLUSIONS: Reduced cerebellar diameter at term equivalent age is related to abnormal GMs at 3months' corrected age, independent of white matter abnormality and intraventricular haemorrhage. PMID- 20004535 TI - Clinically apparent internal mammary nodal metastasis in patients with advanced breast cancer: incidence and local control. AB - PURPOSE: To investigate the incidence and local control of internal mammary lymph node metastases (IMN+) in patients with clinical N2 or N3 locally advanced breast cancer. METHODS AND MATERIALS: We retrospectively reviewed the records of 809 breast cancer patients diagnosed with advanced nodal disease (clinical N2-3) who received radiation treatment at our institution from January 2000 December 2006. Patients were considered IMN+ on the basis of imaging studies. RESULTS: We identified 112 of 809 patients who presented with IMN+ disease (13.8%) detected on ultrasound, computed tomography (CT), positron emission tomography/CT (PET/CT), and/or magnetic resonance imaging (MRI) studies. All 112 patients with IMN+ disease received anthracycline and taxane-based chemotherapy. Neoadjuvant chemotherapy (NCT) resulted in a complete response (CR) on imaging studies of IMN disease in 72.1% of patients. Excluding 16 patients with progressive disease, 96 patients received adjuvant radiation to the breast or the chest wall and the regional lymphatics including the IMN chain with a median dose of 60 Gy if the internal mammary lymph nodes normalized after chemotherapy and 66 Gy if they did not. The median follow-up of surviving patients was 41 months (8-118 months). For the 96 patients able to complete curative therapy, the actuarial 5-year IMN control rate, locoregional control, overall survival, and disease-free survival were 89%, 80%, 76%, and 56%. CONCLUSION: Over ten percent of patients with advanced nodal disease will have IMN metastases on imaging studies. Multimodality therapy including IMN irradiation achieves excellent rates of control in the IMN region and a DFS of more than 50% after curative treatment. PMID- 20004537 TI - On the possible role of mammary-derived growth hormone in human breast cancer. AB - The incidence of breast cancer has risen worldwide, especially in countries where it used to be low, very probably as a result of economic prosperity and changes in life-style. In women, the available data have resulted in the concept of progression from normal breast development to cancer through precursor lesions sensitive to hormones and growth factors that can be produced locally in the mammary gland, acting as paracrine or autocrine stimulating agents. The local endocrine environment in the breast can be different from the situation in the circulation. In the dog, growth hormone (GH) can be produced locally in the mammary glands and its production can be stimulated by progestins. This GH probably plays a paracrine role in the progesterone-induced proliferation and differentiation of mammary epithelium. There is increasing evidence that the local mammary progestin/GH-axis is operational not only in dogs but also in human breast cancer. No data are yet available on the production of mammary-derived GH in women. PMID- 20004538 TI - Waist circumference and serum adiponectin levels in obese and non-obese postmenopausal women. AB - OBJECTIVES: A proposed missing link between obesity and metabolic disturbances is adiponectin, an adipocyte-derived peptide. Adiponectin is a potent antidiabetic hormone and seems to have a beneficial influence on lipid profile as well. The need to explain the complex physiological roles of this hormone prompted the authors to study the relationship between adiponectin level and obesity - related abnormalities in a homogenous population of postmenopausal women. STUDY DESIGN: The study involved 272 postmenopausal women aged 50-60 years. Invitations to participate in the study were sent to 4000 randomly chosen women from the Wroclaw city population fulfilling the age criterion. A telephone questionnaire was administered to the group of 1731 women who responded to the invitation and then subjects for the study were selected. Main outcome measures anthropometrical measurements of body fat tissue content and fat tissue distribution assessment were carried out in all the women. Moreover, serum concentrations of adiponectin, glucose, total cholesterol, HDL cholesterol, triglycerides and insulin were measured. RESULTS: The most frequent (76%) phenotype among the investigated women was obesity (BMI >25) with abnormal (=80cm) waist circumference (OAW), Obesity with normal (<80cm) waist (ONW) and normal weight with abnormal waist (NOAW) were observed in only 5% and 14% of the women, respectively. Non-obese women with normal waist (NONW) were noted in only 5% of the subjects. Serum adiponectin levels in both groups of non-obese women (NOAW and NONW) were significantly higher (p<0.05) than in the women with obesity or overweight and abnormal waist circumference (OAW group). Adiponectin levels in the women with obesity or overweight and normal waist (ONW) were also higher than in the OAW group; however, this difference was not statistically significant (p=0.05). In all the women, serum adiponectin level correlated negatively with BMI (r=-0.34, p=0.0001), total fat (r=-0.28, p=0.0001), android fat deposit (r=-0.23, p=0.0001), waist circumference (r=-0.33, p=0.0001), glucose (r=-0.27, p=0.0001), triglycerides (r=-0.34, p=0.0001), and FIRI (r=-0.34, p=0.0001) and positively with the gynoid/android fat deposit ratio (r=0.28, p=0.0001) and HDL cholesterol (r=0.36, p=0.0001). CONCLUSIONS: These results confirm that adiponectin could be a marker of the development of menopausal insulin resistance syndrome. PMID- 20004539 TI - Prescribing postmenopausal hormone therapy to women in their 50s in the post Women's Health Initiative era. PMID- 20004541 TI - Preparation and solid-state characterization of bupivacaine hydrochloride cyclodextrin complexes aimed for buccal delivery. AB - Binary products of bupivacaine hydrochloride (BVP HCl), an amide type local anesthetic, with parent beta-cyclodextrin (beta-CD) and its soluble beta cyclodextrin-epichlorohydrin polymer (EPI-beta-CD) were prepared and evaluated as a first phase in the development of a novel mucoadhesive formulation aimed for buccal delivery of this drug. The solid products were obtained by physical mixing, ball milling in high-energy mills, co-evaporation and lyophilisation, in order to rationally select the most effective preparation technique. The solid products obtained were carefully characterised by differential scanning calorimetry (DSC), X-ray powder diffractometry (XRPD), Fourier transform infrared spectroscopy (FTIR) and environmental scanning electron microscopy (ESEM). The impact of the preparation techniques on the physicochemical properties of plain drug was also studied. Results of solid-state analysis revealed more intense interactions of BVP HCl with EPI-beta-CD than with native beta-CD, accompanied by stronger reduction of drug crystallinity in the samples, probably favoured by the amorphous nature of the polymeric carrier. While summarising the results of DSC and XRPD analyses, it seems that ball milling of drug/cyclodextrin binary mixtures was particularly efficient in inducing solid-state interaction between the components and it can be considered as the method of choice for preparation of complexes of BVP HCl with beta-CD and EPI-beta-CD. In vitro dissolution properties in artificial saliva of ball-milled BVP HCl and corresponding CD complexes were investigated by simulating the conditions present at the surface of the buccal mucosa. The obtained results confirmed that complexation of BVP HCl with beta-CD and EPI-beta-CD is a suitable tool for properly tailoring the dissolution properties of the drug and it can be favourably exploited for the development of an effective buccal drug delivery system. PMID- 20004542 TI - Simultaneous determination of irbesartan and hydrochlorothiazide in human plasma using HPLC coupled with tandem mass spectrometry: Application to bioequivalence studies. AB - A sensitive, specific and selective liquid chromatography/tandem mass spectrometric method has been developed and validated for the simultaneous determination of irbesartan and hydrochlorothiazide in human plasma. Plasma samples were prepared using protein precipitation with acetonitrile, the two analytes and the internal standard losartan were separated on a reverse phase C(18) column (50mmx4mm, 3microm) using water with 2.5% formic acid, methanol and acetonitrile (40:45:15, v/v/v (%)) as a mobile phase (flow rate of 0.70mL/min). Irbesartan and hydrochlorothiazide were ionized using ESI source in negative ion mode, prior to detection by multiple reaction monitoring (MRM) mode while monitoring at the following transitions: m/z 296-->269 and m/z 296-->205 for hydrochlorothiazide, 427-->175 for irbesartan. Linearity was demonstrated over the concentration range 0.06-6.00microg/mL for irbesartan and 1.00-112.00ng/mL for hydrochlorothiazide. The developed and validated method was successfully applied to a bioequivalence study of irbesartan (300mg) with hydrochlorothiazide (12.5mg) tablet in healthy volunteers (N=36). PMID- 20004543 TI - Simultaneous determination of eleven bioactive compounds in Saururus chinensis from different harvesting seasons by HPLC-DAD. AB - A high performance liquid chromatography method coupled with diode array detection (HPLC-DAD) was developed for simultaneous determination of five major active flavonoids, two aristolactams and four main lignans in Saururus chinensis, namely rutin (1), isoquercitrin (2), quercetin-3-O-beta-d-glucopyranosyl (1-->4) alpha-l-rhamnoside (3), quercitrin (4), quercetin (5), aristolactam A II (6), sauristolactam (7), dihydroguaiaretic acid (8), sauchinone (9), licarin A (10) and licarin B (11). The analysis was performed on an Agilent Eclipse XDB C(18) column (4.6mmx150mm, 5microm) with gradient elution of 0.4% aqueous phosphoric acid and acetonitrile. The detection wavelengths were 280 and 360nm. All calibration curves showed good linearity (r(2)>0.9991) within test ranges. The method was reproducible with intra- and inter-day variation less than 3.2%. The recovery of the assay was in the range of 95.1-103.9%. The validated method was successfully applied for the analysis of the eleven bioactive compounds in seven samples from different harvesting seasons and significant variations were revealed. The results indicated that the developed method can be used as a suitable quality control method for S. chinensis and it should be harvested in August (fruiting period) for Jiangsu cultivation regions, taking the yield into consideration, with the highest amounts of lignans, relative higher amounts of flavonoids and lower amounts of aristolactams. PMID- 20004544 TI - A combined cell membrane chromatography and online HPLC/MS method for screening compounds from Radix Caulophylli acting on the human alpha(1A)-adrenoceptor. AB - We have developed an online analytical method that combines alpha(1A) adrenoceptor (alpha(1A)AR) cell membrane chromatography (alpha(1A)AR-CMC) with high performance liquid chromatography and mass spectrometry (HPLC/MS) for the identification of active components from Radix Caulophylli acting on the human alpha(1A)AR. Fractions retained by the alpha(1A)AR-CMC column were captured into a loop and the components were directly analyzed by combining an 8 port column switcher with an HPLC/MS system for separation and preliminary identification. Using methoxamine as a positive control drug, magnoflorine and caulophine from Radix Caulophylli were identified as the active molecules acting on the alpha(1A)AR. This new alpha(1A)AR-CMC-online-HPLC/MS method can be applied for screening active components acting on alpha(1A)AR from traditional Chinese medicines exemplified by Radix Caulophylli. This method will be of great utility in drug discovery using natural medicinal herbs as a source of novel compounds. PMID- 20004546 TI - Computer-aided detection in computed tomography colonography with full fecal tagging: comparison of standalone performance of 3 automated polyp detection systems. AB - PURPOSE: We sought to compare the performance of 3 computer-aided detection (CAD) polyp algorithms in computed tomography colonography (CTC) with fecal tagging. METHODS: CTC data sets of 33 patients were retrospectively analysed by 3 different CAD systems: system 1, MedicSight; system 2, Colon CAD; and system 3, Polyp Enhanced View. The polyp database comprised 53 lesions, including 6 cases of colorectal cancer, and was established by consensus reading and comparison with colonoscopy. Lesions ranged from 6-40 mm, with 25 lesions larger than 10 mm in size. Detection and false-positive (FP) rates were calculated. RESULTS: CAD systems 1 and 2 could be set to have varying sensitivities with higher FP rates for higher sensitivity levels. Sensitivities for system 1 ranged from 73%-94% for all lesions (78%-100% for lesions > or =10 mm) and, for system 2, from 64%-94% (78%-100% for lesions > or =10 mm). System 3 reached an overall sensitivity of 76% (100% for lesions > or =10 mm). The mean FP rate per patient ranged from 8-32 for system 1, from 1-8 for system 2, and was 5 for system 3. At the highest sensitivity level for all polyps (94%), system 2 showed a statistically significant lower FP rate compared with system 1 (P = .001). When analysing lesions > or =10 mm, system 3 had significantly fewer FPs than systems 1 and 2 (P < .012). CONCLUSIONS: Standalone CTC-CAD analysis in the selected patient collective showed the 3 systems tested to have a variable but overall promising performance with respect to sensitivity and the FP rate. PMID- 20004545 TI - Testing whether decision aids introduce cognitive biases: results of a randomized trial. AB - OBJECTIVE: Women at high risk of breast cancer face a difficult decision whether to take medications like tamoxifen to prevent a first breast cancer diagnosis. Decision aids (DAs) offer a promising method of helping them make this decision. But concern lingers that DAs might introduce cognitive biases. METHODS: We recruited 663 women at high risk of breast cancer and presented them with a DA designed to experimentally test potential methods of identifying and reducing cognitive biases that could influence this decision, by varying specific aspects of the DA across participants in a factorial design. RESULTS: Participants were susceptible to a cognitive bias - an order effect - such that those who learned first about the risks of tamoxifen thought more favorably of the drug than women who learned first about the benefits. This order effect was eliminated among women who received additional information about competing health risks. CONCLUSION: We discovered that the order of risk/benefit information influenced women's perceptions of tamoxifen. This bias was eliminated by providing contextual information about competing health risks. PRACTICE IMPLICATIONS: We have demonstrated the feasibility of using factorial experimental designs to test whether DAs introduce cognitive biases, and whether specific elements of DAs can reduce such biases. PMID- 20004547 TI - The benefits of computed tomographic colonography in reducing a long colonoscopy waiting list. AB - PURPOSE: The Radiology Department, Royal Jubilee Hospital, Victoria, BC, with the support of gastroenterologists and surgeons, was awarded a BC Innovation fund to run a pilot project of computed tomographic colonography to reduce an unacceptably long 2-year colonoscopy waiting list. Funds were approved in April 2007 for a 1-year project, which was completed on March 31, 2008. METHODS: This article describes the challenges of delivering a high-volume computed tomographic colonography program at a busy community hospital, with discussion of the results for the 2,005 patients who were examined. RESULTS: Colonoscopy was avoided in 1,462 patients whose computed tomographic studies showed no significant lesions. In the remainder of patients, only lesions larger than 5 mm were reported, with a total of 508 lesions identified in 433 patients. There were 57 cancers of which 52 were reported as either definite or possible cancers, whereas 5 were not seen on initial scans. Some of the patients with cancer had been on the colonoscopy waiting list for 2 years. In addition, there were 461 patients with significant extracolonic findings, including 84 who required urgent or semi-urgent further management for previously unsuspected conditions, such as pneumonia, aneurysms larger than 5 cm, and a range of solid renal, hepatic, and pancreatic masses. There were no procedural complications from the computed tomographic colon studies. CONCLUSIONS: We have shown that it is feasible to run a high volume CTC service in a general hospital given hospital support and funding. The benefits in this group of over 2000 patients included avoidance of colonoscopy in over 70% of patients, detection of significant polyps or cancer in approximately 20% of patients, and identification of clinically important conditions in 7%-18% depending on the definition used. The estimated costs including capital, operating, and professional fees were in the range of $400. PMID- 20004548 TI - Magnetic resonance cisternographic evaluation of glossopharyngeal, vagus, and accessory nerves. AB - PURPOSE: The individual visualization of the glossopharyngeal, vagus, and accessory nerves has been a troublesome issue. After the recent developments in the microsurgical field, the detailed knowledge of the relationship of these nerves and the tumour has gained importance. The purpose of this study is to compare the visibility of each of these nerves. METHODS: Thirty patients (M/F: 14/16; mean age 52.46 years) with complaints of vertigo, tinnitus, and hearing loss were examined with routine temporal magnetic resonance imaging (MRI) study. The imaging protocol consisted of 3-dimensional fast imaging with steady state acquisition in axial and sagittal oblique planes in addition to routine sequences. These images were transferred to a workstation and reformatted. Visibility of the nerves was evaluated by consensus of 2 radiologists who used an evaluation scale of 2 (excellently visible), 1 (partially visible), to 0 (not visible). RESULTS: In 26 patients, both sides were scanned; in 4 patients, only one side was scanned. A total of 168 nerves were investigated. The rates for visualization for each nerve were as follows: glossopharyngeal nerve, 100% and 100%; vagus nerve, 67.9% and 100%; and accessory nerve, 10.8% and 83.85% on axial and sagittal oblique 3-dimensional fast imaging with steady state acquisition, respectively. CONCLUSIONS: Glossopharyngeal, vagus, and accessory nerve assessment improved when images were obtained in the sagittal oblique plane to the jugular foramen. PMID- 20004549 TI - The use of the Amplatzer plug to treat dysphagia lusoria caused by an aberrant right subclavian artery. AB - Emerging technology with endovascular techniques has expanded our armamentarium to treat the aberrant right subclavian artery. We describe a hybrid technique using an Amplatzer plug in combination with a carotid subclavian bypass to treat a patient with dysphagia lusoria. PMID- 20004550 TI - The Kaiser Permanente inpatient risk adjustment methodology was valid in an external patient population. AB - OBJECTIVES: Accurately predicting hospital mortality is necessary to measure and compare patient care. External validation of predictive models is required to truly prove their utility. This study assessed the Kaiser Permanente inpatient risk adjustment methodology for hospital mortality in a patient population distinct from that used for its derivation. STUDY DESIGN AND SETTING: Retrospective cohort study at two hospitals in Ottawa, Canada, involving all inpatients admitted between January 1998 and April 2002 (n=188,724). Statistical models for inpatient mortality were derived on a random half of the cohort and validated on the other half. RESULTS: Inpatient mortality was 3.3%. The model using original parameter estimates had excellent discrimination (c-statistic 89.4, 95% confidence interval [CI] 0.891-0.898) but poor calibration. Using data based parameter estimates, discrimination was excellent (c-statistic 0.915, 95% CI 0.912-0.918) and remained so when patient comorbidity was expressed in the model using the Elixhauser Index (0.901, 0.898-0.904) or the Charlson Index (0.894, 0.891-0.897). These models accurately predicted the risk of hospital death. CONCLUSION: The Kaiser Permanente inpatient risk adjustment methodology is a valid model for predicting hospital mortality risk. It performed equally well regardless of methods used to summarize patient comorbidity. PMID- 20004551 TI - Allocation concealment continues to be misunderstood. PMID- 20004552 TI - Moving knowledge to action through dissemination and exchange. AB - OBJECTIVE: The objective of this article is to discuss the knowledge dissemination and exchange components of the knowledge translation process that includes synthesis, dissemination, exchange, and ethically sound application of knowledge. This article presents and discusses approaches to knowledge dissemination and exchange and provides a summary of factors that appear to influence the effectiveness of these processes. It aims to provide practical information for researchers and knowledge users as they consider what to include in dissemination and exchange plans developed as part of grant applications. STUDY DESIGN AND SETTING: Not relevant. RESULTS AND CONCLUSIONS: Dissemination is targeting research findings to specific audiences. Dissemination activities should be carefully and appropriately considered and outlined in a dissemination plan focused on the needs of the audience who will use the knowledge. Researchers should engage knowledge users to craft messages and help disseminate research findings. Knowledge brokers, networks, and communities of practice hold promise as innovative ways to disseminate and facilitate the application of knowledge. Knowledge exchange or integrated knowledge translation involves active collaboration and exchange between researchers and knowledge users throughout the research process. PMID- 20004553 TI - Trial sequential analyses of meta-analyses of complications in laparoscopic vs. small-incision cholecystectomy: more randomized patients are needed. AB - OBJECTIVE: Conclusions based on meta-analyses of randomized trials carry a status of "truth." Methodological components may identify trials with systematic errors ("bias"). Trial sequential analysis (TSA) evaluates random errors in meta analysis. We analyzed meta-analyses on laparoscopic vs. small-incision cholecystectomy regarding different outcome measures for the occurrence of type I errors. STUDY DESIGN AND SETTING: Using TSA, we calculated the required information size (IS) and the trial sequential monitoring boundaries regarding complications in our Cochrane review with meta-analyses of cholecystectomy. For each outcome, we calculated a low risk of bias heterogeneity-adjusted IS. As a sensitivity analysis, we calculated an a priori heterogeneity-adjusted IS. RESULTS: According to the trial sequential analyses based on a low risk of bias heterogeneity-adjusted IS definitive evidence may be reached by conducting one more randomized trial. Information may be required on 582 and 119 additional randomized patients to evaluate the effect on severe complications and serious adverse events (SAEs), respectively. CONCLUSION: Our results provide incentives to conduct a new trial with a low risk of bias focusing on a new composite outcome measure of SAEs to obtain conclusive evidence on which operative method to recommend. PMID- 20004554 TI - Patient preferences before and after total knee arthroplasty. AB - OBJECTIVE: Patients before total joint arthroplasty vary in the spectrum and importance of their concerns. The objectives of this study were to evaluate the psychometric properties of the Knee Patient-Specific Index (KPSI) and to determine the type and importance of patients' concerns before and after knee arthroplasty. STUDY DESIGN AND SETTING: A cohort of 119 patients scheduled for elective primary (or revision) total knee arthroplasty were interviewed at two tertiary care teaching hospitals. Patients also completed the Knee Society Scale (KSS), the Short Form 36, the Western Ontario and McMaster University Osteoarthritis Index (WOMAC), and the McMaster Toronto Arthritis Patient Preference Disability Questionnaire (MACTAR). RESULTS: Patients improved after total knee arthroplasty in all 42 symptoms and physical limitations, except crouching/kneeling and walking up and down stairs. Patients' summated concerns correlated with the WOMAC pain subscale (ranging from 0.72 to 0.79), WOMAC physical function subscale (ranging from 0.72 to 0.76), and KSS (ranging from 0.28 to 0.39). The summated responses changed after knee arthroplasty as demonstrated by the standardized response mean of 1.1. CONCLUSIONS: The KPSI captures individual patient unique preferences for patients undergoing total knee arthroplasty. Patients improved in virtually all aspects of their symptoms and function after surgery, with the exception of crouching/kneeling and knee feeling hot. PMID- 20004556 TI - Peri-ictal correlation dynamics of high-frequency (80-200 Hz) intracranial EEG. AB - PURPOSE: To assess (1) how large-scale correlation of intracranial EEG signals in the high-frequency range (80-200Hz) evolves from the pre-ictal, through the ictal into the postictal state and (2) whether the contribution of local neuronal activity to large-scale EEG correlation differentiates epileptogenic from non epileptogenic brain tissue. METHODS: Large-scale correlation of intracranial EEG was assessed by the total correlation strength (TCS), a measure derived from the eigenvalue spectra of zero-lag correlation matrices computed in a time-resolved manner by using a moving window approach. The relative change of total correlation strength (Delta(j)) resulting from leaving out EEG channel j ("leave one-out approach") was used to quantify the contribution of local neuronal activity to large-scale EEG correlation. RESULTS: 19 seizures of 3 patients were analyzed. On average, TCS increased significantly from the pre-ictal to the ictal, and from the ictal to the postictal state. In the pre-ictal state, Delta(j) was significantly more negative when EEG channels that recorded the electrical activity of brain tissue considered to be epileptogenic were left out; the identification of the epileptogenic area, that was subsequently surgically removed in two patients, was based on visual analysis. The spatio-temporal pattern of Delta(j) dramatically changed at seizure onsets and endings, revealing qualitative similarities between the seizures of different patients. DISCUSSION: The evolution of large-scale EEG correlation in the high-frequency range is qualitatively similar to the one previously described for the low-frequency range. Because the two patients who underwent surgery became seizure free, our findings are consistent with the hypothesis that epileptogenic brain tissue may be characterized by its relatively increased contribution to pre-ictal large scale correlation. PMID- 20004555 TI - Pre-existing autoimmunity determines type 1 diabetes outcome after Flt3-ligand treatment. AB - Redirection of immune responses by manipulation of antigen-presenting cells is an emerging strategy for immunosuppressive treatment of autoimmune diseases. In vivo expansion of dendritic cells (DC) by Fms-like tyrosine kinase-3 (Flt3)-Ligand (FL) treatment was shown to delay diabetes onset in the NOD model of autoimmune diabetes. However, we show here that Flt3 stimulation actually accelerates autoimmunity when autoreactive CD8 T cells are detectable in blood prior to treatment. With autoreactive CD8 cells present, the capacity of FL to expand DCs and induce Treg remained intact, but both numbers and the functional response of islet-specific CD8s were boosted. Also, the inhibitory receptor PD-1 on (autoreactive) CD8 T cells and its ligand PD-L1 on Treg were no longer upregulated. These data highlight the need to pre-screen for T cell autoreactivity prior to generalized DC expansion and illustrate how accelerated disease can occur when the intended initiation of regulatory mechanisms is impaired later in diabetogenesis. PMID- 20004557 TI - Isolation and functional characterization of a biosurfactant produced by Lactobacillus paracasei. AB - In this study, the crude biosurfactant produced by a Lactobacillus paracasei strain isolated in a Portuguese dairy industry was characterized. The minimum surface tension (41.8mN/m) and the critical micelle concentration (2.5mg/ml) obtained were found to be similar to the values previously reported for biosurfactants isolated from other lactobacilli. The biosurfactant was found to be stable to pH changes over a range from 6 to 10, being more effective at pH 7, and showed no loss of surface activity after incubation at 60 degrees C for 120h. Although the biosurfactant chemical composition has not been determined yet, a fraction was isolated through acidic precipitation, which exhibited higher surface activity as compared with the crude biosurfactant. Furthermore, this isolated biosurfactant showed antimicrobial and anti-adhesive activities against several pathogenic microorganisms. In addition, L. paracasei exhibited a strong autoaggregating phenotype, which was maintained after washing and resuspending the cells in PBS, meaning that this attribute must be related to cell surface components and not to excreted factors. The autoaggregation ability exhibited by this strain, together with the antimicrobial and anti-adhesive properties observed for this biosurfactant opens the possibility for its use as an effective probiotic strain. PMID- 20004558 TI - Encapsulation of ascorbyl palmitate in chitosan nanoparticles by oil-in-water emulsion and ionic gelation processes. AB - The encapsulation of ascorbyl palmitate (AP) in chitosan particles was carried out by droplet formation via an oil-in-water emulsion, followed by droplet solidification via ionic gelation using sodium triphosphate pentabasic (TPP) as a cross-linking agent. The success of AP encapsulation was confirmed by FT-IR, UV vis spectrophotometry, TGA, and XRD techniques. The obtained AP-loaded chitosan particles were spherical in shape with an average diameter of 30-100nm as observed by SEM and TEM. Loading capacity (LC) and encapsulation efficiency (EE) of AP in the nanoparticles were about 8-20% and 39-77%, respectively, when the initial AP concentration was in the range of 25-150% (w/w) of chitosan. Augmentation of the initial AP concentration led to an increase of LC and a reduction of EE. The amount of AP released from the nanoparticles in ethanol and tris buffer (pH approximately 8.0) increased with increasing LC and decreasing TPP concentration. PMID- 20004559 TI - [Incidence of malpractice claims involving pediatricians]. PMID- 20004560 TI - [Acardiac headless twin: a case report and review of the literature]. AB - INTRODUCTION: Monochorial twin pregnancies are characterised by the presence of vascular connections between the twins. These connections can be the cause of pathologies such as the twin-twin transfusion syndrome or the TRAP syndrome, which is defined as the association of a headless, acardiac twin with a healthy twin. CASE REPORT: The case of an acardiac, headless twin diagnosed during pregnancy at 24 weeks of amenorrhea is described. DISCUSSION: An acardiac headless twin is a rare phenomenon. Yet early diagnosis is crucial during pregnancy to provide adequate monitoring. It is associated with a high death rate in the healthy twin caused by anaemia and heart failure. Therapeutic resources involve interruption of vascular anostomoses between the twins in order to perform a selective feticide. CONCLUSION: Treatment of acardiac headless twin pregnancy ranges from obstetric abstentionism to interventionism, which depends on the prognosis for the healthy twin, dominated by the risk of preterm birth and heart failure. PMID- 20004561 TI - Smooth-to-striated muscle transition in human esophagus: an immunohistochemical study using fetal and adult materials. AB - BACKGROUND: A craniocaudal transition from smooth to striated muscle occurs in the fetal mouse esophagus muscularis propria, until finally the entire muscle component becomes striated. Although no such investigation has been conducted using human fetuses, the transition appears to be incomplete. METHODS: In horizontal sections of 10 human fetuses between 9 and 16 weeks of gestation, we identified immunoreactivity for smooth muscle actin (SMA), striated muscle myosin heavy chain (MyH), desmin, PGP9.5, S100 protein, c-kit, and CD68 in the thoracic esophagus. The TUNEL method was used to identify apoptosis. For comparison, the same immunohistochemistry was conducted using 10 adult esophaguses. RESULTS: In fetuses at all stages examined, a transition zone was found in the upper thoracic esophagus that was attached to the middle one-third of the trachea. In the transition zone, the MyH-positive longitudinal muscle fibers were surrounded by flat, SMA-positive cells, whereas the MyH-positive circular fibers were sometimes located adjacent to the SMA-positive fibers. However, in adults, smooth muscle tended to be clearly separated from striated muscle. The distribution of cells showing immunoreactivity for PGP9.5, S100 or c-kit did not differ between the oral and anal sides of the transition zone. Desmin was positive in the muscularis propria, but negative in the muscularis mucosae. Neither CD68-positive macrophages nor TUNEL-positive cells were present in the esophagus. CONCLUSIONS: In the human esophagus, the smooth-to-striated muscle transition appears to stop at the mid-thoracic level. Cell death or transdifferentiation of smooth muscle appears unlikely, but phenotypic transformation into desmin-positive myofibroblasts is a possibility. PMID- 20004562 TI - Hypertonic environment elicits cyclooxygenase-2-driven prostaglandin E2 generation by colon cancer cells: role of cytosolic phospholipase A2-alpha and kinase signaling pathways. AB - Cyclooxygenase (COX)-2-derived prostaglandin (PG)E(2) controls many aspects of colon cancer development, modulating from apoptosis resistance and cell proliferation to angiogenesis, invasion, and metastasis. Here, we investigated the role of different phospholipases (PL)A(2) in supplying arachidonic acid (AA) for COX-2-dependent PGE(2) generation and signaling pathways involved in activation of colon cancer cells by a physiologically relevant stimulus. To emulate the hypertonic environment found physiologically in colon, the human colon cancer cell line Caco-2 was maintained in hypertonic complete DMEM medium. Human colon cancer cell line Caco-2 exposed to a hypertonic environment responded with marked AA release, COX-2 induction and PGE(2) generation. Selective secretory (s)PLA(2) and calcium-independent (i)PLA(2) inhibitors did not modify PGE(2) generation, while either COX-2 or cytosolic (c)PLA(2) inhibitors completely inhibited PGE(2) generation. cPLA(2)-alpha was responsible for AA supply for PGE(2) generation, but had no role in COX-2 induction. Mitogen activated protein (MAP) kinases, ERK 1/2, p38, and JNK, participated in the signaling events that lead to PGE(2) generation by modulating AA release, but only ERK 1/2 was involved in COX-2 upregulation. Our results indicate that hypertonic stress activates PGE(2) generation by Caco-2 cells through a mechanism dependent on MAP kinase-regulated AA mobilization, increased cPLA(2)-alpha activity, and COX-2 induction. PMID- 20004563 TI - A new approach to reduce number of split fields in large field IMRT. AB - Intensity-modulated radiation therapy (IMRT) has been applied for treatments of primary head with neck nodes, lung with supraclavicular nodes, and high-risk prostate cancer with pelvis wall nodes, all of which require large fields. However, the design of the Varian multileaf collimator requires fields >14 cm in width to be split into 2 or more carriage movements. With the split-field technique, both the number of monitor units (MUs) and total treatment time are significantly increased. Although many different approaches have been investigated to reduce the MU, including introducing new leaf segmentation algorithms, none have resulted in widespread success. In addition, for most clinics, writing such algorithms is not a feasible solution, particularly with commercial treatment planning systems. We introduce a new approach that can minimize the number of split fields and reduce the total MUs, thereby reducing treatment time. The technique is demonstrated on the Eclipse planning system V7.3, but could be generalized to any other system. PMID- 20004564 TI - Non-receptor tyrosine kinase Etk regulation of drug resistance in small-cell lung cancer. AB - Epithelial and endothelial tyrosine kinase (Etk), also known as Bmx (bone marrow X kinase), plays an important role in the apoptosis of epithelial cells. The aim of this study was to investigate whether Etk is involved in the chemoresistance of small cell lung cancer (SCLC) and to correlate the drug resistance associated proteins such as bcl-2, bcl-X(L) and p53. Drug-resistant small lung cancer cells (H69AR) which were originally developed by ADM and which demonstrated multi-drug resistance to chemotherapeutic agents were used in the study. Western blot analysis revealed that H69AR cells over-expressed the proteins Etk and bcl-X(L), but not bcl-2 and p53 when compared to parent H69 cells. Knockdown of Etk expression by Etk-specific small interfering RNA sensitised H69AR cells to chemotherapeutic drugs and inhibited bcl-X(L) expression but not bcl-2 and p53. Co-immunoprecipitation was performed to further evaluate the relationship between Etk and bcl-X(L) with anti-Etk and anti-phospho-Etk antibodies. The bcl-X(L) was accompanied with a robust increase of Etk and tyrosine phosphorylated Etk at Tyr 40 in H69AR cells. In conclusion, our results suggest that non-receptor tyrosine kinase Etk is involved in drug resistance to SCLC by mediating bcl-X(L) via Tyr(P)-40. The potential approach for downregulation of Etk activity on expression would be a novel, potentially clinically practical strategy for interfering with chemoresistance in SCLC. PMID- 20004565 TI - COX-2 expression in chondrosarcoma: a role for celecoxib treatment? AB - Chondrosarcomas are resistant to conventional chemo- and radiotherapy. A subset of chondrosarcomas arises secondarily in the benign tumour syndromes enchondromatosis (EC) and multiple osteochondromas (MO), and prevention of tumour development would greatly improve prognosis. We therefore investigated the effect of selective COX-2 inhibition on chondrosarcoma growth. COX-2 expression was studied in central- and peripheral cartilaginous tumours. The effect of COX-2 inhibition was assessed in four high-grade chondrosarcoma cell lines using celecoxib and NS-398 treatment. COX-2 activity (prostaglandin E(2) (PGE(2)) ELISA) and cell viability were measured. The (prophylactic) effect of celecoxib on chondrosarcoma growth in vivo was studied for 8 weeks using a xenograft model of cell line CH2879 in immunoincompetent nude mice. High COX-2 protein expression was mainly found in solitary peripheral chondrosarcoma and in enchondromatosis related central chondrosarcoma, which was confirmed by qPCR. After 72h of celecoxib treatment, a significant decrease in cell viability was observed in three chondrosarcoma cell lines. In vivo, celecoxib initially slowed tumour growth in chondrosarcoma xenografts; however, after prolonged treatment relapsed tumour growth was observed. Tumour volume was negatively associated with celecoxib serum levels, and seemed smaller in the high-dose prophylactic treatment group. We confirmed the expression of COX-2 in 65% of chondrosarcomas, and COX-2 inhibition by celecoxib diminished cell viability in vitro. The initial response and the decrease in tumour volume with increased celecoxib serum levels in vivo supported a role for celecoxib, although relapsed tumour growth after 6 weeks was worrisome. Also the role of high-dose prophylactic celecoxib in preventing the development of benign and malignant cartilage tumours in EC and MO patients deserves further investigation. PMID- 20004566 TI - Age-related physiologic changes and perioperative management of elderly patients. AB - Aging itself only minimally increases operative risk, but with aging, there is an increase of the prevalence of chronic diseases and a progressive deterioration of organ function. Aging is associated with a progressive decrease in heart, lung and kidney performance. Under normal conditions, these physiologic changes do not produce any problems for the elderly, but when these patients are subjected to the stress of surgery or its complications, there may be inadequate functional reserve. It is very important to know age-related patho-physiological changes in order to be able to better evaluate elderly patients undergoing surgery, and to prevent and manage preoperative complications. In this review we try to identify and to describe the most frequent physiological changes in the elderly, how those impact pharmacodynamic and pharmacokinetic parameters, and how to assess and manage them. PMID- 20004567 TI - Isolation and characterization of wheat straw lignin with a formic acid process. AB - There is a commercial need to increase the amount of industrial produced lignin. An 86.24 wt.% formic acid solution with 0.2M H(2)SO(4) as the catalyst was used to isolate the lignin of dewaxed wheat straw at different residence times and reaction temperatures. Lignin was extracted and purified separately from hydrolysates and residual solids after treatment. The lignin obtained has been characterized by FTIR (Fourier transform infrared), GPC (Gel permeation chromatography), and (1)H NMR (Nuclear magnetic resonance). Results from these analyses indicated that oxidative cleavage of bonds in the lignin macromolecule has occurred during the formic acid treatment, leading to formation of oligomers of lignin with lower molecular weight. Moreover, it was found that guaiacyl unit is the main structure, the content of syringyl unit is higher than that of the hydroxybenzyl unit and beta-O-4 together with beta-5 ether bonds is also identified in the molecular structure of lignin of wheat straw obtained from the formic acid process. PMID- 20004568 TI - High concentration ethanol production from corncob residues by fed-batch strategy. AB - Ethanol production from corncob residues (CCR) pretreated by different methods was studied. The structure features of these CCR were analyzed by Fourier transform-infrared spectrum (FT-IR), X-ray diffraction (XRD), and field emission scanning electron microscope (SEM). Simultaneous saccharification and fermentation (SSF) was performed by adding crude cellulase preparations from Penicillium decumbens JUA10-1 at 30 degrees C. The results suggested that different pretreatments resulted in different composition and structure of residues; these changes had a significant influence on ethanol productivity and concentration. The fed-batch method was combined with SSF to enhance ethanol concentration further and reduce enzyme dosage. Moreover, the absorption and desorption phenomena of cellobiohydrolase I (CBH I) (70 kDa) were observed to be related to lignin contents in residues. These results demonstrated that despite the application of low enzyme dosage, high concentration ethanol could be produced from pretreated corncobs by combining fed-batch method with SSF. PMID- 20004569 TI - Production, characterization and application of activated carbon from brewer's spent grain lignin. AB - Different types of activated carbon were prepared by chemical activation of brewer's spent grain (BSG) lignin using H(3)PO(4) at various acid/lignin ratios (1, 2, or 3g/g) and carbonization temperatures (300, 450, or 600 degrees C), according to a 2(2) full-factorial design. The resulting materials were characterized with regard to their surface area, pore volume, and pore size distribution, and used for detoxification of BSG hemicellulosic hydrolysate (a mixture of sugars, phenolic compounds, metallic ions, among other compounds). BSG carbons presented BET surface areas between 33 and 692 m(2)/g, and micro- and mesopores with volumes between 0.058 and 0.453 cm(3)/g. The carbons showed high capacity for adsorption of metallic ions, mainly nickel, iron, chromium, and silicon. The concentration of phenolic compounds and color were also reduced by these sorbents. These results suggest that activated carbons with characteristics similar to those commercially found and high adsorption capacity can be produced from BSG lignin. PMID- 20004571 TI - Inhibition of CTP synthase from Escherichia coli by xanthines and uric acids. AB - CTP synthase (CTPS) catalyzes the conversion of UTP to CTP and is a recognized target for the development of anticancer, antiviral, and antiprotozoal agents. Xanthine and related compounds inhibit CTPS activity (IC(50)=0.16-0.58mM). The presence of an 8-oxo function (i.e., uric acids) enhances inhibition (IC(50)=0.060-0.121mM). An intact purine ring with anionic character favors inhibition. In general, methylation of the purine does not significantly affect inhibition. PMID- 20004570 TI - P4 capped amides and lactams as HCV NS3 protease inhibitors with improved potency and DMPK profile. AB - SAR studies on the extension of P3 unit of Boceprevir (1, SCH 503034) with amides and lactams and their synthesis is described. Extensive SAR studies resulted in the identification of 36 bearing 4, 4-dimethyl lactam as the new P4 cap unit with improved potency (K(i)( *)=15nM, EC 90=70nM) and pharmacokinetic properties (Rat AUC (PO)=3.52microMh) compared to 1. PMID- 20004572 TI - Synthesis and PGE(2) production inhibition of 1H-furan-2,5-dione and 1H-pyrrole 2,5-dione derivatives. AB - 3,4-Diphenyl-substituted 1H-furan-2,5-dione and 1H-pyrrole-2,5-dione derivatives were synthesized and evaluated for the inhibitory activities on LPS-induced PGE(2) production in RAW 264.7 macrophage cells. Both 1H-furan-2,5-dione and 1H pyrrole-2,5-dione rings as main scaffolds were easily obtained using one of three synthetic methods. Among the compounds investigated, 1H-3-(4-sulfamoylphenyl)-4 phenyl-pyrrole-2,5-dione (6l) showed a strong inhibitory activity (IC(50)=0.61microM) of PGE(2) production. PMID- 20004574 TI - Parallel synthesis of N-biaryl quinolone carboxylic acids as selective M(1) positive allosteric modulators. AB - An iterative analog library synthesis approach was employed in the exploration of a quinolone carboxylic acid series of selective M(1) positive allosteric modulators, and strategies for improving potency and plasma free fraction were identified. PMID- 20004573 TI - Investigation of the sequence and length dependence for cell-penetrating prenylated peptides. AB - Cell penetrating peptides are useful delivery tools for introducing molecules of interest into cells. A new class of cell penetrating molecules has been recently reported-cell penetrating, prenylated peptides. In this study a series of such peptides was synthesized to examine the relationship between peptide sequence and level of peptide internalization and to probe their mechanism of internalization. This study revealed that prenylated peptides internalize via a non-endocytotic pathway regardless of sequence. Sequence length and identity was found to play a role in peptide uptake but prenylated sequences as short as two amino acids were found to exhibit significant cell penetrating properties. PMID- 20004575 TI - Antitubercular potential of some semisynthetic analogues of phytol. AB - Phytol, a diterpene alcohol was modified to several semisynthetic analogues. Some of the modifications were done logically to enhance lipophilicity of the molecule. Analogues 14, 16 and 18 exhibited antitubercular activity (MIC 15.6 50microg/mL) better than phytol (100microg/mL). The most potent analogue 18 was evaluated for in vivo toxicity in Swiss albino mice and was well tolerated by the experimental animals up to 300mg/kg body weight as a single oral acute dose. PMID- 20004577 TI - Evaluation of human neutrophil elastase inhibitory effect of iridoid glycosides from Hedyotis diffusa. AB - Five iridoid glycosides were isolated from the MeOH extract of Hedyotis diffusa, and their structures were elucidated as E-6-O-p-methoxycinnamoyl scandoside methyl ester (1), Z-6-O-p-methoxycinnamoyl scandoside methyl ester (2), E-6-O-p feruloyl scandoside methyl ester (3), E-6-O-p-coumaroyl scandoside methyl ester (4), and Z-6-O-p-coumaroyl scandoside methyl ester (5) by interpretation of their spectroscopic data. All the isolated compounds were evaluated for human neutrophil elastase inhibitory effect, and compound 1 showed potent activity with an IC(50) value of 18.0muM. The molecular docking simulation suggested a structural model for the inhibition of human neutrophil elastase by compound 1. PMID- 20004576 TI - Non-peptide entry inhibitors of HIV-1 that target the gp41 coiled coil pocket. AB - The ectodomain of HIV-1 gp41 mediates the fusion of viral and host cellular membranes. The peptide-based drug Enfuvirtide(1) is precedent that antagonists of this fusion activity may act as anti HIV-agents. Here, NMR screening was used to discover non-peptide leads against this target and resulted in the discovery of a new benzamide 1 series. This series is non-peptide, low molecular weight, and analogs have activity in a cell fusion assay with EC50 values ranging 3-41microM. Structural work on the gp41/benzamide 1 complex was determined by NMR spectroscopy using a designed model peptide system that mimics an open pocket of the fusogenic form of the protein. PMID- 20004578 TI - Chemical lead optimization of a pan G(q) mAChR M(1), M(3), M(5) positive allosteric modulator (PAM) lead. Part I: Development of the first highly selective M(5) PAM. AB - This Letter describes a chemical lead optimization campaign directed at VU0238429, the first M(5)-preferring positive allosteric modulator (PAM), discovered through analog work around VU0119498, a pan G(q) mAChR M(1), M(3), M(5) PAM. An iterative library synthesis approach delivered the first selective M(5) PAM (no activity at M(1)-M(4) @ 30microM), and an important tool compound to study the role of M(5) in the CNS. PMID- 20004580 TI - An anatomical assessment of the supracerebellar midline and paramedian approaches to the inferior colliculus for auditory midbrain implants using a neuronavigation model on cadaveric specimens. AB - The inferior colliculus (IC) is an alternative site for electrode placement in neural deafness due to its surgical accessibility and its well-known tonotopic stratification. In patients where tumor surgery has already occurred and the cerebellopontine angle contains scar tissue or tumor-remnants, midline and paramedian supracerebellar approaches are alternative routes. They are often avoided due to concerns regarding the venous drainage of the cerebellum, the electrode trajectory and the course of the electrode cable. We studied these surgical routes in five neuronavigated fixed cadaveric specimens. For paramedian and midline approaches, the transverse sinus was exposed 5.8mm on average. A mean of 1.6 cerebellar veins, with an average diameter of 2.0mm, draining to the tentorium were transected to reach the tentorial notch. Only 0.4 arterial branches were met. We conclude that the supracerebellar midline and paramedian approaches provide a good exposure of the IC and offer safe and viable alternative routes to the IC. Additionally, they provide a wider angle of action for optimal electrode placement. PMID- 20004579 TI - Nuclear networking fashions pre-messenger RNA and primary microRNA transcripts for function. AB - The expression of protein-coding genes is enhanced by the exquisite coupling of transcription by RNA polymerase II with pre-messenger RNA processing reactions, such as 5'-end capping, splicing and 3'-end formation. Integration between cotranscriptional processing events extends beyond the nucleus, as proteins that bind cotranscriptionally can affect the localization, translation and degradation of the mature messenger RNA. MicroRNAs are RNA polymerase II transcripts with crucial roles in the regulation of gene expression. Recent data demonstrate that processing of primary microRNA transcripts might be yet another cotranscriptional event that is woven into this elaborate nuclear network. This review discusses the extensive molecular interactions that couple the earliest steps in gene expression and therefore influence the final fate and function of the mature messenger RNA or microRNA produced. PMID- 20004581 TI - Optic nerve atrophy in adrenoleukodystrophy detectable by optic coherence tomography. AB - The adrenoleukodystrophies (ALDs) are a group of metabolic disorders characterised by the accumulation of very long-chain fatty acids in all tissues. The two most frequent ALD phenotypes are adult-onset adrenomyeloneuropathy (AMN) and childhood cerebral ALD. Visual system involvement in the adult phenotype is well described as impairment of visual function and optic disc pallor on clinical examination accompanied by demyelination of the optic nerves seen on MRI. Thinning of the retinal nerve fiber layer and ganglion cell death has been described in a neonatal form of ALD. Our patient provides evidence, through ocular coherence tomography scanning of the retina, that such degenerative changes also underlie the visual dysfunction seen in the AMN phenotype. PMID- 20004582 TI - Comparative PET study using F-18 FET and F-18 FDG for the evaluation of patients with suspected brain tumour. AB - The aim of this prospective pilot study in patients with suspected or known brain tumour was to establish the diagnostic value of O-(2-[(18)F]-fluoroethyl)-L tyrosine (FET) positron emission tomography (PET) when compared to fluorine-18 fluorodeoxyglucose (FDG) PET. Twenty-five FET PET and FDG PET scans were performed on 21 consecutive patients within 24 months. Final malignant pathology included 11 glioma (eight low-grade, three high grade), two lymphoma, one olfactory ganglioneuroblastoma, one anaplastic meningioma. Benign pathology included two encephalitis and one cortical dysplasia. Definitive pathology was not available in three patients. The accuracy of PET was determined by subsequent surgical histopathology in 12 and clinical/imaging course in nine patients. Median follow-up period was 20 months. FET sensitivity was 93%, specificity 100%, accuracy 96%, positive predictive value (PPV) 100% and negative predictive value (NPV) 91%. FDG sensitivity was 27%, specificity 90%, accuracy 52%, PPV 80% and NPV 45%. FET PET is more accurate than FDG PET for detecting malignant brain lesions, especially low-grade gliomas. PMID- 20004583 TI - Physiological dimensions of ototoxic responses in a model fish species. AB - Pharmaceutical agents known to be toxic to the human auditory system also impair sensory hair cells of teleosts, and this supports the use of fish models for the screening of such compounds. However, previous investigations have focused almost exclusively on anatomical changes after drug administration without assessing macro-level physiological effects. Using the goldfish (Carassius auratus), we demonstrate that the acquisition of auditory evoked potentials offers a rapid and non-invasive means for tracking ototoxin-induced shifts in auditory thresholds. Gentamicin (100mg/mL) was the agent of choice as it is an extensively-studied human ototoxin. Significant shifts (p<0.05) in hearing sensitivity were observed between 300 Hz and 600 Hz and these shifts depended on acoustic pressure, but not particle motion. This differential elevation of auditory thresholds may be caused by impairment of specific populations of auditory sensory hair cells. PMID- 20004584 TI - 3-(2-Aminocarbonylphenyl)propanoic acid analogs as potent and selective EP3 receptor antagonists. Part 1: discovery and exploration of the carboxyamide side chain. AB - A series of 3-(2-aminocarbonyl-4-phenoxymethylphenyl)propanoic acid analogs were synthesized and evaluated for their EP3 antagonist activity in the presence of additive serum albumin. Several compounds were biologically evaluated for their in vivo efficacy with respect to the PGE(2)-induced uterine contraction in pregnant rats as well as their pharmacokinetics. The discovery process of these potent and selective EP3 antagonists and their structure activity relationship are also presented. PMID- 20004585 TI - Synthesis, anti-HIV and anti-oxidant activities of caffeoyl 5,6-anhydroquinic acid derivatives. AB - In our continued research on chlorogenic acid analogues and derivatives with improved bioactivity, we have synthesized some caffeoyl 5,6-anhydroquinic acid derivatives. The 1,7 acetonides of chlorogenic acid (15), and of the mono caffeoyl 5,6-anhydroquinic acids (7-8) showed appreciable anti-HIV activity. The 3,4-dicaffeoyl 5,6-anhydroquinic acid (12) exhibited an anti-HIV activity twice as that of 3,5-dicaffeoylquinic acid (22). The caffeoyl 5,6-anhydroquinic acid derivatives displayed potent anti-oxidant activities. The mono-caffeoyl 5,6 anhydroquinic acids (10-11) were more than twice stronger than chlorogenic acid (21) on SOD-like activity. PMID- 20004586 TI - Structural and ultrastructural characteristics of male reproductive tract and spermatozoa in two Cryptinae species (Hymenoptera: Ichneumonidae). AB - This study represents the first characterization of male reproductive tracts and ultrastructural description of sperm of Cryptinae species. In Lymeon dieloceri and Pachysomoides sp., the male reproductive tract is formed by a pair of testis, two deferent ducts, two accessory glands and one ejaculatory duct. The spermatozoa are similar to those described for other Hymenoptera, with: (1) the acrosome formed by the acrosomal vesicle covering the perforatorium, which has its base inserted in a cavity located in the nucleus point; (2) a thin nucleus with electron-dense chromatin; (3) an electron-dense centriolar adjunct located between the nucleus and one of the two mitochondrial derivatives; (4) an axoneme with a 9 + 9 + 2 microtubule arrangement; (5) two long mitochondrial derivatives with peripheral cristae and; (6) two accessory bodies located between the two mitochondrial derivatives and the axoneme. These ichneumonids present structural characteristics similar to other parasitic wasps, such as presence of a single follicle per testis, layer of extracellular material enveloping the acrosome and accessory microtubules ending before others in final portion of the flagellum. However, male reproductive system and the spermatozoa presented morphological characteristics that allowed their differentiation, such as oval shapes accessory glands and the symmetric mitochondrial derivatives in L. dieloceri compared to the spherical accessory glands and asymmetrical derivatives observed in Pachysomoides sp. Taken together, data presented here demonstrates that diversity of morphological characteristic from the male reproductive tract and spermatozoa in Hymenoptera might provide a character system that can be used, in association with other systems, to resolve various uncertainties about the evolutionary relationships of this insect group. PMID- 20004587 TI - Determination of 51Cr and 241Am X-ray and gamma-ray emission probabilities per decay. AB - In this paper results of X-ray and gamma-ray emission probabilities per decay of (51)Cr and (241)Am are presented. The measurements were carried out by means of HPGe planar and REGe spectrometers. The activity of (51)Cr and (241)Am samples was determined in a 4pibeta-gamma coincidence counting system. The HPGe spectrometers were calibrated in a well defined geometry by means of (54)Mn, (55)Fe, (57)Co, (133)Ba, (152)Eu, (166m)Ho and (241)Am sources, previously standardized in a 4pibeta-gamma coincidence system. The MCNP Monte Carlo code was used for simulation of the REGe spectrometer calibration curve, for the selected geometry, and compared with the experimental curve. The experimental results were compared with data from literature. PMID- 20004588 TI - IFIN-HH ionization chamber calibration and its validation; electrometric system improvement. AB - The paper presents the results obtained in the calibration of the CENTRONIC IG12/20A ionization chamber for 18 gamma ray emitters, and its improvement with a Keithley 6517A electrometer. The calibration figures were determined either directly in pA MBq(-1) units, or calculated from old units, by using the system capacity value. The calibration figures, determined with RML's standards, are compared with those deduced from the KCRV or the mean of the comparisons, and the values determined at PTB-Germany. PMID- 20004589 TI - Analysis of selective androgen receptor modulators by gas chromatography microchip atmospheric pressure photoionization-mass spectrometry. AB - A gas chromatography-microchip atmospheric pressure photoionization-mass spectrometric (GC-microAPPI-MS) method was developed and used for the analysis of three 2-quinolinone-derived selective androgen receptor modulators (SARMs). SARMs were analyzed from spiked urine samples, which were hydrolyzed and derivatized with N-methyl-N-(trimethylsilyl)trifluoroacetamide before analysis. Trimethylsilyl derivatives of SARMs formed both radical cations (M(+*)) and protonated molecules ([M + H](+)) in photoionization. Better signal-to-noise ratios (S/N) were obtained in MS/MS analysis using the M(+*) ions as precursor ions than using the [M + H](+) ions, and therefore the M(+*) ions were selected for the precursor ions in selected reaction monitoring (SRM) analysis. Limits of detection (LODs) with the method ranged from 0.01 to 1 ng/mL, which correspond to instrumental LODs of 0.2-20 pg. Limits of quantitation ranged from 0.03 to 3 ng/mL. The mass spectrometric response to the analytes was linear (R > or = 0.995) from the LOQ concentration level up to 100 ng/mL concentration, and intra day repeatabilities were 5%-9%. In addition to the GC-microAPPI-MS study, the proof-of-principle of gas chromatography-microchip atmospheric pressure chemical ionization-Orbitrap MS (GC-microAPCI-Orbitrap MS) was demonstrated. PMID- 20004590 TI - The origins of the Rag genes--from transposition to V(D)J recombination. AB - The recombination activating genes 1 and 2 (Rag1 and Rag2) encode the key enzyme that is required for the generation of the highly diversified antigen receptor repertoire central to adaptive immunity. The longstanding model proposed that this gene pair was acquired by horizontal gene transfer to explain its abrupt appearance in the vertebrate lineage. The analyses of the enormous amount of sequence data created by many genome sequencing projects now provide the basis for a more refined model as to how this unique gene pair evolved from a selfish DNA transposon into a sophisticated DNA recombinase essential for immunity. PMID- 20004591 TI - Cognitive-neuroscience approaches to issues of philosophy-of-mind. PMID- 20004592 TI - Recovery of shoulder strength and proprioception after open surgery for recurrent anterior instability: a comparison of two surgical techniques. AB - BACKGROUND: Previous studies have documented a decrease in proprioceptive capacity in the unstable shoulder. The degree to which surgical approach affects recovery of strength and proprioception is unknown. MATERIALS AND METHODS: The recovery of strength and proprioception after open surgery for recurrent anterior glenohumeral instability was compared for 2 surgical procedures. A prospective analysis of 55 consecutive patients with posttraumatic unilateral recurrent anterior glenohumeral instability was performed. Thirty patients (group 1) underwent an open inferior capsular shift with detachment of the subscapularis, and 25 (group 2) underwent an anterior capsulolabral reconstruction. RESULTS: Mean preoperative proprioception and strength values were significantly lower for the affected shoulders in both groups. At 6 months after surgery, there were no significant differences for mean strength and proprioception values between the unaffected and operative sides for group 2 patients. In group 1 patients, however, there were still significant deficits in mean position sense and strength values. Complete restoration of proprioception and strength, however, was evident by 12 months in group 1. CONCLUSION: This study demonstrates that there are significant deficits in both strength and proprioception in patients with posttraumatic, recurrent anterior glenohumeral instability. Although both are completely restored by 1 year after surgery, a subscapularis-splitting approach allows for complete recovery of strength and position sense as early as 6 months postoperatively. Detachment of the subscapularis delays recovery of strength and position sense for up to 12 months after surgery. PMID- 20004593 TI - The effect of posterior capsular tightening on peak subacromial contact pressure during simulated active abduction in the scapular plane. AB - HYPOTHESIS: Our hypothesis was that tightening of the posterior capsule would lead to increased subacromial pressure and increased superior translation during active abduction in the scapular plane. BACKGROUND: Subacromial impingement syndrome is a painful condition that occurs during overhead activities as the rotator cuff is compressed in the subacromial space. Unrecognized secondary causes of subacromial impingement may lead to treatment failure. Posterior capsular tightness, believed to alter glenohumeral joint kinematics, is often cited as a secondary cause of SI; however, scientific evidence is lacking. The primary objective of this study was to evaluate the effect of posterior capsular tightening on peak subacromial pressure during abduction in the scapular plane. MATERIALS AND METHODS: Ten fresh frozen shoulder specimens from deceased donors were mounted on a custom shoulder simulator. With the scapula fixed, the deltoid and rotator cuff muscles were loaded in discrete static steps with a constant ratio to elevate the humerus in the scapular plane. The treatment order (no tightening, 1-cm, and 2-cm tightening of the posterior capsule) was randomly assigned to each specimen. Peak subacromial contact pressure and glenohumeral kinematics at the peak pressure position were compared using a repeated measures analysis of variance. RESULTS: Peak subacromial pressures (mean +/- standard deviation) were similar between treatment groups: 345 +/- 152, 410 +/- 213, and 330 +/- 164 kPa for no tightening, 1-cm, and 2-cm tightening of the posterior capsule respectively (P > .05). No significant differences were found for superior or anterior translations at the peak pressure position (P > .05). DISCUSSION: Posterior capsular tightening, as a sole variable, did not contribute to a significant increase in peak subacromial pressure during abduction in the scapular plane. A similar study simulating active forward flexion is necessary to fully characterize the contribution of posterior capsular tightness to subacromial impingement. CONCLUSION: Tightening of the posterior capsule did not increase subacromial pressure, or increase superior or anterior translation during abduction in the scapular plane. PMID- 20004594 TI - Surgical treatment for lateral epicondylitis: a long-term follow-up of results. AB - HYPOTHESIS: Since its first description, the pathology, natural history, and treatment of lateral epicondylitis have remained controversial. For patients in who conservative management fails, surgery remains an option. The optimal method of surgery remains debatable and is further confounded by a relative lack of long term follow-up studies. MATERIALS AND METHODS: This study describes a modification of the Nirschl surgical technique and presents its long-term results. Patients undergoing this open technique were reviewed by use of the Hospital for Special Surgery and Mayo elbow performance assessment tools, as well as having grip strength and subjective outcome recorded. RESULTS: From June 1986 to December 2001, 158 consecutive patients (171 elbows) underwent surgery in a single-surgeon series. Of these patients, 137 (86.7%) were available for follow up at a mean of 9.8 years. The mean age of the group was 42 years. Subjectively, the results were good to excellent in 94.6% of patients and in 92.6% to 94.0% of patients by use of the Hospital for Special Surgery and Mayo scores, respectively. No differences were noted in grip strength. No patient required revision surgery. CONCLUSIONS: This repeatable open technique offers excellent results with a low rate of complications at a mean follow-up of 9.8 years. These results compare favorably in terms of numbers followed up, length of follow-up, and outcome and offer strong evidence of its efficacy. PMID- 20004596 TI - ATP-sensitive potassium channels contribute to the time-dependent alteration in the pentylenetetrazole-induced seizure threshold in diabetic mice. AB - Although there is evidence that diabetes affects seizure susceptibility, the underlying mechanism has not been completely understood. Several studies also suggest a pivotal role for K(ATP) channels in the seizure modulation. The aim of the present study was to evaluate the seizure threshold induced by pentylenetetrazole in diabetic mice at different times (3 days, 1-8 weeks) after induction of diabetes with streptozocin and to examine the possible role of ATP sensitive potassium (K(ATP)) channels in this manner. Our data showed a time dependent alteration in the threshold in diabetic mice, reaching a peak on week 2 after streptozocin injection and declining significantly afterwards. The seizure threshold in 8-week diabetic mice was even lower than control levels, though the difference was not significant. The K(ATP) channel opener cromakalim (0.1 30microg/kg, i.p.) significantly increased the seizure threshold in control mice. Although the K(ATP) channel blocker glibenclamide (0.5, 1mg/kg) had no effect, it prevented the effects of the potent dose of cromakalim (30microg/kg) on seizure threshold in control mice. Glibenclamide (1mg/kg, i.p.) also decreased the seizure threshold in 2-week diabetic mice to the control levels which was blocked by pre-treatment with cromakalim (10microg/kg, i.p.). Cromakalim (10microg/kg, i.p.) significantly increased the seizure threshold in 8-week diabetic mice which was inhibited by pre-treatment with glibenclamide (1mg/kg, i.p.). We demonstrated a time-dependent alteration in the pentylenetetrazole-induced seizure threshold in diabetic mice. This phenomenon might be due to the probable alteration in the K(ATP) channel functioning during the diabetic condition. PMID- 20004595 TI - Arthroscopic capsular release for refractory shoulder stiffness: a critical analysis of effectiveness in specific etiologies. AB - HYPOTHESIS: The purpose of this study is to report and compare the outcome of arthroscopic capsular release in patients with shoulder stiffness with post traumatic, postsurgical, and idiopathic etiologies. We hypothesize that patients with idiopathic or post-traumatic stiffness have better outcomes after arthroscopic capsular release than those with shoulder stiffness with a postsurgical etiology. MATERIALS AND METHODS: A retrospective review of 115 patients who underwent arthroscopic capsular release for refractory shoulder stiffness was performed. There were 60 men and 55 women with a mean age of 49 years (range, 27 to 81 years). The patients were divided into 3 groups according to the etiology of stiffness: post-traumatic (26 patients), postsurgical (48 patients), and idiopathic (41 patients). Arthroscopic capsular release was performed in all patients after a mean of 9 months of physical therapy (range, 6 to 13 months). RESULTS: At a mean follow-up of 46 months (range, 25 to 89 months), the overall subjective shoulder value in all groups improved from 29% to 73% and the age- and gender-adjusted Constant score improved from 35% to 86%. The mean pain score decreased from 7.5 to 1, and mean active forward flexion, external rotation, and internal rotation increased from 97 degrees , 14 degrees , and the L5 vertebral level, respectively, to 135 degrees , 38 degrees , and the T11 vertebral level, respectively (P < .0001). There was no significant difference between the outcomes of idiopathic and post-traumatic stiffness (P = .7). However, the Constant score and subjective shoulder value were significantly lower in the postsurgical group compared with the idiopathic and post-traumatic groups (P = .0001 and P = .006, respectively). CONCLUSIONS: Arthroscopic capsular release is an effective treatment for refractory shoulder stiffness. Patients with idiopathic and post-traumatic shoulder stiffness have better outcomes than patients with postsurgical stiffness. PMID- 20004597 TI - Diffusion weighted MR imaging of the breast. AB - The aim of this work is to review the techniques and clinical applications of diffusion-weighted magnetic resonance (MR) imaging of the breast. Diffusion weighted MR imaging plays a role in the differentiation breast cancer from benign lesions, the characterization of malignancy, and the detection of tumor extension. The apparent diffusion coefficient of breast cancer is correlated with tumor cellularity and some prognostic factors of breast cancer. It can be used for the differentiation of recurrent tumors from posttreatment changes and monitoring of patients after chemotherapy. Diffusion-weighted MR imaging is used for the characterization of breast mass, diagnosis, and the grading and staging of breast cancer, as well as prediction of the responses of patients with breast cancer to chemotherapy. PMID- 20004598 TI - Dissemination of radiological information using enhanced podcasts. AB - RATIONALE AND OBJECTIVES: Podcasts and vodcasts (video podcasts) have become popular means of sharing educational information via the Internet. MATERIALS AND METHODS: In this article, we introduce another method, an enhanced podcast, which allows images to be displayed with the audio. Bookmarks and URLs may also be imbedded within the presentation. This article describes a step-by-step tutorial for recording and distributing an enhanced podcast using the Macintosh operating system. Enhanced podcasts can also be created on the Windows platform using other software. An example of an enhanced podcast and a demonstration video of all the steps described in this article are available online at web.mac.com/mthapa. RESULTS: An enhanced podcast is an effective method of delivering radiological information via the Internet. Viewing images while simultaneously listening to audio content allows the user to have a richer experience than with a simple podcast. Incorporation of bookmarks and URLs within the presentation will make learning more efficient and interactive. CONCLUSION: The use of still images rather than video clips equates to a much smaller file size for an enhanced podcast compared to a vodcast, allowing quicker upload and download times. PMID- 20004599 TI - Correlation of CSF proinflammatory cytokines with MRI in tuberculous meningitis. AB - RATIONALE AND OBJECTIVES: To demonstrate the correlation of proinflammatory cytokines (PCs), intercellular adhesion molecule (sICAM-1), tumor necrosis factor alpha (TNF-alpha), and interleukin-1beta (IL-1beta) in CSF of tuberculous meningitis (TBM) patients with magnetic resonance imaging (MRI) including diffusion tensor imaging (DTI) and also to look for the changes in imaging parameters after antitubercular treatment (ATT) in these patients. MATERIALS AND METHODS: Forty patients with TBM (median age, 27.7 years) and 30 age-/sex-matched controls were included in this study. PCs were quantified from the CSF of TBM patients at the time of hospital admission (baseline). MRI including DTI was performed at the time of baseline study and 6 months after ATT. RESULTS: Significant positive correlation of PCs with fractional anisotropy (FA) values and post-contrast signal intensity (PCSI) collected from cerebral cortical regions was observed in TBM patients. A significant positive correlation of FA values with PCSI was also observed at both time points in patient groups. At baseline study significantly high FA values were observed in patients compared to controls. Significantly decreased FA values and PCSI were observed in the patients after 6 months of ATT compared to the baseline study. CONCLUSIONS: Results of this study suggest that the DTI-derived anisotropy have the potential to delineate meningeal inflammation and it may be used in assessment of therapeutic response in TBM patients as an additional method to conventional imaging. PMID- 20004600 TI - Structural analysis of two novel mutations in MCFD2 gene causing combined coagulation factors V and VIII deficiency. PMID- 20004601 TI - Subarachnoid transplantation of immortalized galanin-overexpressing astrocytes attenuates chronic neuropathic pain. AB - Treatment of chronic neuropathic pain resulted from peripheral nerve injury is one of the most difficult problems in modern clinical practice. The use of cell lines as biologic "minipumps" to chronically deliver anti-nociceptive molecules into the pain-processing centers of spinal cord is a newly developing technique for the treatment of pain. Moreover, spinal administration of exogenous galanin (GAL) is a useful target for the treatment of chronic pain after nerve injury. Because of better histocompatibility, lower immunogenicity and reproducibility, immortalized astrocytes (IAST) have been served as a promising cellular vehicle to deliver therapeutic molecules into CNS. In this study, the rat IAST was transfected with rat preprogalanin cDNA and the galanin-synthesizing and secreting cell line, IAST/GAL, was isolated. After cells were transplanted into the subarachnoid space of rats with chronic neuropathic pain induced by spared nerve injury (SNI) of sciatic nerve, their analgesic potential was evaluated by behavioral tests. The results showed that IAST/GAL transfected with preprogalanin gene could express and secrete significantly higher level of GAL protein in vitro and in vivo as compared with control cells. In addition, the pain-related behaviors, thermal hyperalgesia and mechanical allodynia were significantly alleviated during the 1-7 weeks after grafts of IAST/GAL cells, which could be reversed by galanin receptor antagonist M35 temporarily. Taken together, these data suggest that subarachnoid transplant of immortalized galanin-overexpressing astrocytes near the pain-processing centers was able to reverse the development of chronic neuropathic pain, which offers an adjunct approach to currently used therapies for the pain management. PMID- 20004603 TI - icoshift: A versatile tool for the rapid alignment of 1D NMR spectra. AB - The increasing scientific and industrial interest towards metabonomics takes advantage from the high qualitative and quantitative information level of nuclear magnetic resonance (NMR) spectroscopy. However, several chemical and physical factors can affect the absolute and the relative position of an NMR signal and it is not always possible or desirable to eliminate these effects a priori. To remove misalignment of NMR signals a posteriori, several algorithms have been proposed in the literature. The icoshift program presented here is an open source and highly efficient program designed for solving signal alignment problems in metabonomic NMR data analysis. The icoshift algorithm is based on correlation shifting of spectral intervals and employs an FFT engine that aligns all spectra simultaneously. The algorithm is demonstrated to be faster than similar methods found in the literature making full-resolution alignment of large datasets feasible and thus avoiding down-sampling steps such as binning. The algorithm uses missing values as a filling alternative in order to avoid spectral artifacts at the segment boundaries. The algorithm is made open source and the Matlab code including documentation can be downloaded from www.models.life.ku.dk. PMID- 20004602 TI - SEnD NMR: sensitivity enhanced n-dimensional NMR. AB - Sparse sampling offers tremendous potential for overcoming the time limitations imposed by traditional Cartesian sampling of indirectly detected dimensions of multidimensional NMR data. However, in many instances sensitivity rather than time remains of foremost importance when collecting data on protein samples. Here we explore how to optimize the collection of radial sampled multidimensional NMR data to achieve maximal signal-to-noise. A method is presented that exploits a rigorous definition of the minimal set of radial sampling angles required to resolve all peaks of interest in combination with a fundamental statistical property of radial sampled data. The approach appears general and can achieve a substantial sensitivity advantage over Cartesian sampling for the same total data acquisition time. Termed Sensitivity Enhanced n-Dimensional or SEnD NMR, the method involves three basic steps. First, data collection is optimized using routines to determine a minimal set of radial sampling angles required to resolve frequencies in the radially sampled chemical shift evolution dimensions. Second, appropriate combinations of experimental parameters (transients and increments) are defined by simple statistical considerations in order to optimize signal-to noise in single angle frequency domain spectra. Finally, the data is processed with a direct multidimensional Fourier transform and a statistical artifact and noise removal step is employed. PMID- 20004604 TI - In silico and functional studies of the regulation of the glucocerebrosidase gene. AB - In Gaucher disease (GD), the inherited deficiency of glucocerebrosidase results in the accumulation of glucocerebroside within lysosomes. Although almost 300 mutations in the glucocerebrosidase gene (GBA) have been identified, the ability to predict phenotype from genotype is quite limited. In this study, we sought to examine potential GBA transcriptional regulatory elements for variants that contribute to phenotypic diversity. Specifically, we generated the genomic sequence for the orthologous genomic region ( approximately 39.4kb) encompassing GBA in eight non-human mammals. Computational comparisons of the resulting sequences, using human sequence as the reference, allowed the identification of multi-species conserved sequences (MCSs). Further analyses predicted the presence of two putative clusters of transcriptional regulatory elements upstream and downstream of GBA, containing five and three transcription factor-binding sites (TFBSs), respectively. A firefly luciferase (Fluc) reporter construct containing sequence flanking the GBA gene was used to test the functional consequences of altering these conserved sequences. The predicted TFBSs were individually altered by targeted mutagenesis, resulting in enhanced Fluc expression for one site and decreased expression for seven others sites. Gel-shift assays confirmed the loss of nuclear-protein binding for several of the mutated constructs. These identified conserved non-coding sequences flanking GBA could play a role in the transcriptional regulation of the gene contributing to the complexity underlying the phenotypic diversity seen in GD. PMID- 20004605 TI - Tumor necrosis factor-alpha-308A allele may have a protective effect for chronic hepatitis B virus infection in Mongoloid populations. AB - OBJECTIVES: Previous studies on the tumor necrosis factor-alpha (TNF-alpha)-308 gene promoter polymorphism in chronic hepatitis B virus (HBV) infection have reported conflicting results. METHODS: We carried out a meta-analysis of 21 studies in relation to the TNF-alpha-308 gene promoter, involving a total of 4230 chronic HBV infection cases and 2905 controls. RESULTS: The overall meta-analysis indicated that -308A heterozygotes (GA) had a significant 27% decreased risk of developing chronic hepatitis B (CHB) (odds ratio (OR) 0.73; 95% confidence interval (CI) 0.57-0.93; p=0.012). For -308A allele homozygotes (AA) and carriers (GA+AA), the pooled odd ratios both indicated a significantly decreased risk of CHB (OR 0.28; 95% CI 0.19-0.43; p=0.0001; and OR 0.70; 95% CI 0.55-0.89; p=0.004, respectively). In subgroup analyses by ethnicity, a significantly decreased risk was associated with -308 variant genotypes (GA and AA) in Mongoloid populations in all genetic models. However, no significant associations were found in Caucasoids. Moreover, in the subgroup analyses by control group, significantly decreased risk was associated with -308 variant genotypes (GA and AA) in the group of spontaneously recovered cases in all genetic models; however, no significant associations were found in the group of healthy cases. CONCLUSIONS: The TNF-alpha-308A allele is a protective factor for chronic HBV infection, especially in Mongoloids. PMID- 20004606 TI - Rural maternity care: can we learn from Wal-Mart? AB - In many countries rural maternity care is under threat. Consequently rural pregnant women will have to travel further to attend larger maternity units to receive care and deliver their babies. This trend is not dissimilar from the disappearance of other rural services, such as village shops, banks, post offices and bus services. We use a comparative approach to draw an analogy with large scale supermarkets, such as the Wal-Mart and Tesco and their effect on the viability of smaller rural shops, depersonalisation of service and the wider community. The closure of a community-maternity unit leads to women attending a different type of hospital with a different approach to maternity care. Thus small community-midwifery units are being replaced, not by a very similar unit that happens to be further away, but by a larger obstetric unit that operates on different models, philosophy and notions of risk. Comparative analysis allows a fresh perspective on the provision of rural maternity services. We argue that previous discussions focusing on medicalisation and change in maternity services can be enhanced by drawing on experience in other sectors and taking a wider societal lens. PMID- 20004607 TI - Automated detection of intracranial aneurysms based on parent vessel 3D analysis. AB - The detection of brain aneurysms plays a key role in reducing the incidence of intracranial subarachnoid hemorrhage (SAH) which carries a high rate of morbidity and mortality. The majority of non-traumatic SAH cases is caused by ruptured intracranial aneurysms and accurate detection can decrease a significant proportion of misdiagnosed cases. A scheme for automated detection of intracranial aneurysms is proposed in this study. Applied to the segmented cerebral vasculature, the method detects aneurysms as suspect regions on the vascular tree, and is designed to assist diagnosticians with their interpretations and thus reduce missed detections. In the current approach, the vessels are segmented and their medial axis is computed. Small regions along the vessels are inspected and the writhe number is introduced as a new surface descriptor to quantify how closely any given region approximates a tubular structure. Aneurysms are detected as non-tubular regions of the vascular tree. The geometric assumptions underlying the approach are investigated analytically and validated experimentally. The method is tested on 3D-rotational angiography (3D-RA) and computed tomography angiography (CTA). In our experiments, 100% sensitivity was achieved with average false positives rates of 0.66 per study on 3D-RA data and 5.36 false positive rates per study on CTA data. PMID- 20004609 TI - Effect of acidity on microstructure and spectroscopic properties of Y2O3:Eu3+ powders and ceramics. AB - Y(2)O(3):Eu(3+) nano-powders were synthesized by homogeneous precipitation and the influence of solution acidity was investigated. IR and TG-DTA analysis showed that in different acidic solutions different kinds of sediments were generated. TEM images revealed distinct morphologies of obtained powders. The resultant powders were sintered into ceramics in vacuum and then in N(2) atmosphere without any additives. SEM images indicated that the morphologies of powders have significant impact on the microstructures of sintering-derived ceramics. Low agglomerated and uniform powders were in favor of production of ceramics with homogeneous microstructures. Excitation and emission spectra of both powders and ceramics were measured and some changes were observed. PMID- 20004608 TI - Reliability of cortical activity during natural stimulation. AB - Response reliability is complementary to more conventional measurements of response amplitudes, and can reveal phenomena that response amplitudes do not. Here we review studies that measured reliability of cortical activity within or between human subjects in response to naturalistic stimulation (e.g. free viewing of movies). Despite the seemingly uncontrolled nature of the task, some of these complex stimuli evoke highly reliable, selective and time-locked activity in many brain areas, including some regions that show little response modulation in most conventional experimental protocols. This activity provides an opportunity to address novel questions concerning natural vision, temporal scale of processing, memory and the neural basis of inter-group differences. PMID- 20004610 TI - Spectrometric studies on the interaction of fluoroquinolones and bovine serum albumin. AB - The interaction between fluoroquinolones (FQs), ofloxacin and enrofloxacin, and bovine serum albumin (BSA) was investigated by fluorescence and UV-vis spectroscopy. It was demonstrated that the fluorescence quenching of BSA by FQ is a result of the formation of the FQ-BSA complex stabilized, in the main, by hydrogen bonds and van der Waals forces. The Stern-Volmer quenching constant, K(SV), and the corresponding thermodynamic parameters, DeltaH, DeltaS and DeltaG, were estimated. The distance, r, between the donor, BSA, and the acceptor, FQ, was estimated from fluorescence resonance energy transfer (FRET). The effect of FQ on the conformation of BSA was analyzed with the aid of UV-vis absorbance spectra and synchronous fluorescence spectroscopy. Spectral analysis showed that the two FQs affected the conformation of the BSA but in a different manner. Thus, with ofloxacin, the polarity around the tryptophan residues decreased and the hydrophobicity increased, while for enrofloxacin, the opposite effect was observed. PMID- 20004611 TI - Analysis of changes in attenuated total reflection FTIR fingerprints of Pseudomonas fluorescens from planktonic state to nascent biofilm state. AB - Attenuated total reflection Fourier transform infrared (ATR-FTIR) spectroscopy is a useful method for monitoring biofilm in situ, non-destructively, in real time, and under fully hydrated conditions. In this work we focused on changes in Pseudomonas fluorescens ATR-FTIR fingerprint accompanying the very early stages of biofilm formation: initial bacterial adhesion and the very beginning of biofilm development in the presence of nutrients. To help interpreting variations in the ATR-FTIR fingerprint of sessile bacteria, ATR-FTIR spectra of planktonic bacteria in different growth phases were also examined, and the average surface coverage and spatial arrangement of bacteria on the ATR crystal were determined by epifluorescence microscopy. The proteins, nucleic acids and polysaccharides ATR-FTIR spectral data recorded during growth of sessile bacteria were shown to be linked to changes in the physiological state of the bacteria, possibly accompanied by extracellular polymeric substances production. This work clearly showed by spectroscopic method how bacteria change drastically their metabolism during the first hours of biofilm formation. PMID- 20004612 TI - Intermanual transfer of sensorimotor memory for grip force when lifting objects: the role of wrist angulation. AB - OBJECTIVE: To investigate the mechanisms underlying the intermanual transfer of sensorimotor memory when lifting an object. METHODS: Twenty healthy subjects grasped and lifted an object with constant mechanical properties with the right hand (RH) first and then with the left hand (LH). Ten of the subjects lifted the object with the RH in a regular wrist angulation (WA), followed by lifts with the LH in a regular WA. The remaining 10 subjects lifted the object with the RH in a hyper-flexed WA, followed by lifts with the LH in a regular WA. RESULTS: Subjects generated greater peak grip force (GF) rates, grip and lift forces when lifting the object with the wrist in a regular WA compared to lifts with the wrist in hyper-flexion. Importantly, subjects transferred the predictive scaling of GF from the RH to the LH, regardless of the WA. CONCLUSIONS: Biomechanical properties of the object do not seem to be used by the CNS as a first line information to evaluate GF when handling an object or transferring information about the grasp to the opposite hemisphere. SIGNIFICANCE: The predictive scaling of GF rather reflects an internal sense of effort than an internal representation of the mechanical object properties. PMID- 20004613 TI - Inhibition of the anterior intraparietal area and the dorsal premotor cortex interfere with arbitrary visuo-motor mapping. AB - OBJECTIVE: The contribution of the human anterior intraparietal area and the dorsal premotor cortex to arbitrary visuo-motor mapping during grasping were tested. METHODS: Trained right-handed subjects reached for and pincer-grasped a cube with the right hand in the absence of visual feedback after the cube location had been displayed for 200ms. During the reaching movements, the colour of the cube changed and visual feedback about the change of colour was provided for 100ms at 500ms after movement onset (at the time of peak grasp aperture). Depending on colour, subjects were instructed to either pincer-grasp the cube in a horizontal or vertical grasp position with the latter necessitating wrist rotation (experiment 1) or to pincer-grasp and transport the cube to either a left or right target position (experiment 2). Within two consecutive 200ms time windows (TMS 1 and 2) starting 500ms and 700ms after movement onset, respectively, double pulses of supra-threshold transcranial magnetic stimulation (inter-stimulus interval: 100ms) were delivered over (i) the left primary motor cortex (90 degrees vertically angulated coil position, control stimulation), (ii) the left dorsal premotor cortex or (ii) the left anterior intraparietal area. RESULTS: Compared to control stimulation, stimulation of the anterior intraparietal area, but not of the dorsal premotor cortex, at TMS 1 delayed the times to wrist rotation (experiment 1) and hand transport (experiment 2). Compared to control stimulation, stimulation of the dorsal premotor cortex, but not of the anterior intraparietal area, at TMS 2 delayed both wrist rotation (experiment 1) and hand transport (experiment 2). CONCLUSIONS: We contend that the anterior intraparietal area and the dorsal premotor cortex are both involved albeit at different phases during the mapping of arbitrary visual cues with goal directed grasp and transport movements. SIGNIFICANCE: These data add to the current understanding of how human cortical areas work in concert during manual activities. PMID- 20004614 TI - Caveats when studying motor cortex excitability and the cortical control of movement using transcranial magnetic stimulation. PMID- 20004615 TI - Putative facilitators and barriers for adherence to CPAP treatment in patients with obstructive sleep apnea syndrome: a qualitative content analysis. AB - INTRODUCTION: Effective treatment of obstructive sleep apnea syndrome (OSAS) with continuous positive airway pressure (CPAP) can reduce morbidity and mortality, but adherence rates are low without a clear consensus of causes. OBJECTIVE: To explore the experiences of adherence to CPAP treatment in patients with OSAS. METHODS: A qualitative content analysis was employed. Data were collected by in depth interviews with 23 purposively selected patients. RESULTS: Adherence to CPAP treatment was summarized according to "facilitators" and "barriers" to CPAP treatment. Facilitators for adherence, as described by the patients were a desire to avoid symptoms, knowledge about the risk for medical consequences, fear of negative social consequences and disturbing the sleep of significant others. Other facilitators were a positive attitude to CPAP treatment, trust in healthcare personnel, a sense of engagement from the spouse and a feeling of physical improvement. Barriers included experiencing practical problems, negative psychological effects of the equipment, and negative attitudes to the treatment. Other barriers were side-effects as well as insufficient support from healthcare personnel and the spouse. CONCLUSION: Adherence to CPAP treatment is a multifaceted problem including patient, treatment, condition, social and healthcare related factors. Knowledge about facilitators and barriers for adherence to CPAP treatment can be used in interventional strategies. PMID- 20004616 TI - The big sports events--how does sports medicine and sports science contribute? PMID- 20004617 TI - Ipilimumab monotherapy in patients with pretreated advanced melanoma: a randomised, double-blind, multicentre, phase 2, dose-ranging study. AB - BACKGROUND: Ipilimumab is a human monoclonal antibody that blocks cytotoxic T lymphocyte antigen 4 and has shown promising activity in advanced melanoma. We aimed to ascertain the antitumour efficacy of ipilimumab in patients with advanced melanoma. METHODS: We undertook a randomised, double-blind, phase 2 trial in 66 centres from 12 countries. 217 patients with previously treated stage III (unresectable) or stage IV melanoma were randomly assigned a fixed dose of ipilimumab of either 10 mg/kg (n=73), 3 mg/kg (n=72), or 0.3 mg/kg (n=72) every 3 weeks for four cycles (induction) followed by maintenance therapy every 3 months. Randomisation was done with a permuted block procedure, stratified on the basis of type of previous treatment. The primary endpoint was best overall response rate (the proportion of patients with a complete or partial response, according to modified WHO criteria). Efficacy analyses were done by intention to treat, whereas safety analyses included patients who received at least one dose of ipilimumab. This study is registered with ClinicalTrials.gov, number NCT00289640. FINDINGS: The best overall response rate was 11.1% (95% CI 4.9-20.7) for 10 mg/kg, 4.2% (0.9-11.7) for 3 mg/kg, and 0% (0.0-4.9) for 0.3 mg/kg (p=0.0015; trend test). Immune-related adverse events of any grade arose in 50 of 71, 46 of 71, and 19 of 72 patients at doses of 10 mg/kg, 3 mg/kg, and 0.3 mg/kg, respectively; the most common grade 3-4 adverse events were gastrointestinal immune-related events (11 in the 10 mg/kg group, two in the 3 mg/kg group, none in the 0.3 mg/kg group) and diarrhoea (ten in the 10 mg/kg group, one in the 3 mg/kg group, none in the 0.3 mg/kg group). INTERPRETATION: Ipilimumab elicited a dose-dependent effect on efficacy and safety measures in pretreated patients with advanced melanoma, lending support to further studies at a dose of 10 mg/kg. FUNDING: Bristol-Myers Squibb. PMID- 20004618 TI - Technological approaches to the scientific explorations of epilepsy and behavior. Editorial/commentary. PMID- 20004619 TI - Working memory abnormalities in chronic interictal epileptic psychosis and schizophrenia revealed by magnetoencephalography. AB - Working memory (WM) deficits are considered a core cognitive dysfunction in schizophrenia. To determine cognitive abnormalities in chronic interictal psychosis (CIP), and to assess whether these abnormalities are distinguishable from those seen in schizophrenia in terms of WM deficits, we used magnetoencephalography during a WM task performed by patients with CIP, nonpsychotic epilepsy, and schizophrenia and by healthy subjects. Multiple Source Beamformer and Brain-Voyager were used for analysis. In both patients with CIP and those with schizophrenia, we found dorsolateral prefrontal hyperactivation and left inferior temporal hypoactivation, as indicated by alpha event-related desynchronization and synchronization, respectively. Patients with schizophrenia also showed alpha2 event-related desynchronization in the mid-prefrontal cortex relative to healthy controls. Direct comparison of patients with CIP and schizophrenia rendered no difference in source-power changes. Our findings indicate similar functional cognitive abnormalities in CIP and schizophrenia in the prefrontal and left temporal cortex, which supports the possibility that these disorders share common underlying pathophysiological mechanisms. PMID- 20004620 TI - Intravenous immunoglobulin therapy in intractable childhood epilepsy: open-label study and review of the literature. AB - Our aim was to investigate the long term effectiveness of intravenous immunoglobulin (IVIG) against intractable childhood epilepsy in the era of new antiepileptics and to determine the predictors of a favorable response in a prospective open-label add-on study. Of thirty-seven 9.9+/-0.9-year-old patients (11 with partial seizures, 26 with generalized seizures of whom 9 had West syndrome and 17 Lennox-Gastaut syndrome) followed for 15+/-3 months, 43% had a >50% decrease in seizures (including 15% seizure free, 229+/-58 compared with 104+/-3 seizures/month, P=0.035: generalized 246+/-318 to 117+/-200, P=0.025, partial 191+/-437 to 72+/-179, P>0.05; power=0.2). Males were more likely to respond than females (P=0.011, odds ratio=9.3). Review of the literature revealed nine other articles reporting efficacy of IVIG against epileptic seizures. Only one other used statistical methods and, unlike ours, showed only a trend toward seizure frequency reduction without achieving statistical significance, presumably because it was underpowered. These results indicate large-scale controlled studies of IVIG in epilepsy are still needed. PMID- 20004621 TI - Choosing a treatment for cervicogenic headache: when? what? how much? PMID- 20004622 TI - Determination of the in vivo posterior loading environment of the Coflex interlaminar-interspinous implant. AB - BACKGROUND CONTEXT: The in vivo loading environment of load-bearing implants is generally largely unknown. Loads are typically approximated from cadaver tests or biomechanical calculations for the preclinical assessment of a device's safety and efficacy. PURPOSE: To determine the actual in vivo loading environment of an elastic interlaminar-interspinous implant (Coflex). STUDY DESIGN: A retrospective radiographic study to noninvasively measure the in vivo implant loads of 176 patients. METHODS: For this study, neutral, flexion, and extension radiographs were quantitatively analyzed using validated image analysis technology. The angle between the Coflex arms was measured for each radiograph and statistically evaluated. Separately, the Coflex implant was characterized using mechanical test data and finite element analysis, which resulted in a load-deformation formula that describes the implant load as a function of its size and elastic deformation. Using the formula and the elastic implant deformation data obtained from the radiographic analysis, the exact implant load was calculated for each patient and each posture. For statistical analysis, the patients were grouped by indication and procedure, which resulted in 12 different groups. The determined loads were compared with the strength of the posterior lumbar spinal elements obtained from the literature and with the static and dynamic mechanical limits of the Coflex interlaminar-interspinous implant. RESULTS: The force data were independent of implant size, diagnosis (with one exception), number of levels of the decompression procedure, number of levels of implantations (one or two), and follow-up time. The median compressive force acting on the Coflex implant was found to be 45.8 N. The maximum load change between flexion and extension was 140 N; the maximum overall load exceeded 239 N in extension. CONCLUSIONS: The average loads exerted by the Coflex implant on the spinous process and lamina are 11.3% and 7.0% of their respective static failure load. The implant fatigue strength is significantly higher than the measured median force, which explains the extremely rare observation of a Coflex fatigue failure. PMID- 20004623 TI - Takotsubo cardiomyopathy and congenital long QT syndrome in a patient with a novel duplication in the Per-Arnt-Sim (PAS) domain of hERG1. PMID- 20004624 TI - Diagnostic value of bipolar precordial leads in Brugada syndrome: more accurate, more simple, or more theoretical? PMID- 20004625 TI - Anodic oxidation of 3,4-dihydroxyphenylacetic acid on carbon electrodes in acetic acid solutions. AB - The electrochemical oxidation of 3,4-dihydroxyphenylacetic acid (DOPAC) on a carbon fiber microelectrode (CF) and a glassy carbon macroelectrode (GC) in glacial acetic acid solutions was investigated using voltammetric techniques. Voltammograms recorded at these electrodes show well-defined single waves or peaks. The proposed mechanism of the anodic oxidation of DOPAC consists of two successive one-electron one-proton steps. The loss of the first electron proceeds irreversibly and determines the overall rate of the electrode process. This stage is accompanied by the generation of an unstable phenoxyl radical in position 4 of the aromatic ring. The second stage of the electrode reaction produces substituted orto-quinone as the final product of the electrode process of DOPAC. DOPAC exhibits more antioxidative power than synthetic BHT and can be useful in food protection against reactive oxygen species. The results presented can help to explain biochemical and antioxidative properties of DOPAC in a living cell and can be useful in determination of this compound in real samples. PMID- 20004626 TI - A voltammetric Rhodotorula mucilaginosa modified microbial biosensor for Cu(II) determination. AB - It is the first report about the usage of Rhodotorula mucilaginosa as a biomaterial to construct a microbial biosensor based on carbon paste for determination of copper. Cu(II) was preconcentrated electrode surface at open circuit and then detected with electrochemical techniques, including Cyclic Voltammetry (CV) and Differential Pulse Stripping Voltammetry (DPSV). Some parameters such as pH of preconcentration solution, preconcentration time, scan rate and effect of interfering heavy metal ions were carried out for optimum responses. The best defined cathodic peak was obtained at pH 5 with 0.05 M NaNO(3) and a scan rate of 100 mV/s. The linear range for the developed microbial biosensor was found in the range of 1.0 x 10(-7) and 1.0 x 10(-5)M (0.0064 and 0.64 mg/L) at the response time of 15 min (R(2)=0.98). The easy fabrication, sensitivity, low cost and fast response time showed the advantages of the biosensor to conventional techniques. PMID- 20004627 TI - Capillary electrophoresis of chitooligosaccharides in acidic solution: simple determination using a quaternary-ammonium-modified column and indirect photometric detection with crystal violet. AB - Five chitosan oligosaccharides were separated in acidic aqueous solution by capillary electrophoresis (CE) with indirect photometric detection using a positively coated capillary. Electrophoretic mobility of the chitooligosaccharides (COSs) depended on the number of monomer units in acidic aqueous solution, similar to other polyelectrolyte oligomers. The separation was developed in nitric acid aqueous solution at pH 3.0 with 1 mM Crystal Violet, using a capillary positively coated with N-trimethoxypropyl-N,N,N trimethylammonium chloride. The limit of the detection for chitooligosaccharides with two to six saccharide chains was less than 5 microM. CE determination of an enzymatically hydrolyzed COS agreed with results from HPLC. PMID- 20004629 TI - [Hyperglycaemic hyperosmolar non-ketotic syndrome: a rare onset of diabetes in childhood]. PMID- 20004628 TI - Direct binding of sulfur mustard and chloroethyl ethyl sulphide to human cell membrane-associated proteins; implications for sulfur mustard pathology. AB - Sulfur mustard (SM) is a potent vesicating agent that produces debilitating blisters and ulcerating lesions on the skin which are characteristically slow to heal. There are currently no specific medical countermeasures to prevent SM induced vesication and therefore SM remains a major military threat. To investigate the mechanism by which SM causes these injuries we aimed to identify the cellular proteins that are important in the vesicant response and pathology of SM. Membrane and membrane-associated proteins that are targets for direct binding by SM were compared to targets directly bound by CEES (chloroethylethylsulphide). As CEES is a less potent blistering agent compared to SM, it was hypothesised that differences in the binding of these two mustards could reveal key proteins directly involved in the mustard vesicant response. Human cellular membranes fractionated from HaCaT cells were exposed to (14)C-SM or (14)C-CEES and the membrane proteins to which SM or CEES bound were separated by 2D gel electrophoresis, located by fluorography and subsequently identified using mass spectrometry. A number of proteins were identified that were differentially labelled by SM and CEES. Actin, annexin A2 and keratin 9 were labelled with SM at a higher intensity than was seen with the same concentration of CEES. Therefore results from these studies suggest that SM binding to these proteins could contribute to the complex pathology seen following SM exposure. PMID- 20004630 TI - Surgical resection of vulva lymphoedema circumscriptum. AB - BACKGROUND: Lymphangioma circumscriptum, a rare, benign disease that can be either congenital or acquired, involves the deep dermis and subcutaneous lymphatics. OBJECTIVE: This study aims to analyse the efficacy of surgical resection of vulva lymphangioma circumscriptum (VLC). MATERIALS AND METHODS: Between January 2000 and December 2008, eight consecutive women referred to our centre and treated surgically for VLC were included in the study. VLC was responsible for recurrent lymph oozing in seven cases. All women were treated by the same plastic surgeon specialising in lymphatic diseases. RESULTS: The first surgery was performed after a median interval of 5.4 years since VLC onset. The first cutaneous resection included the labia majora of all women and labia minora of five and clitoral hood of four. Five women experienced rapidly recurrent vesicles associated with lymph oozing and underwent resection again (once: two women, twice: three women). The second resection was performed 4-6 months after the first, whereas the third took place 1-6 years after the second. Five women had moderate and transitory post-surgical lymph oozing. After a median follow-up of 53 months after the last surgery, seven of the eight women were free from symptom. CONCLUSION: Surgical resection is an effective and well-tolerated therapy for VLC in most women. Lesion recurrence is frequent but resection can be repeated several times with no adverse effects. PMID- 20004631 TI - Management of wounds with exposed bone structures using an artificial dermis and skin grafting technique. AB - The task of managing an open wound complicated by exposed bony structures underneath is difficult, if not challenging. We have instituted a method of managing the problems in stages using an artificial dermis and skin grafting technique in 17 wounds in 15 individuals from Sept. 2006 to Feb. 2009. While all wounds were noted to assume aberrant healing processes, the majority of involved bony structures were devoid of periosteal covering compounded by various degrees of infection. Of 15 incidents, mechanical trauma was responsible for 10, chemical burns for two and electrical burns for two patients. A chronic non-healing ulcer with exposed bone formed in an old burn scar accounted for the remaining one. The regimen of surgical management consisted of initial debridement, the coverage of the resultant wound with an artificial dermis and a partial-thickness skin grafted over this dermis-like structure grown with granulation tissues. Complete wound healing was attained in 15 out of 17 with outstanding cosmetic and minimal donor-site morbidity. Despite initial failure encountered in two, the morbidities noted were low. It is especially useful in large defects that usually require flaps for coverage. PMID- 20004632 TI - Microsurgery training today and future. PMID- 20004633 TI - Maternal perceptions of infant hunger, satiety, and pressuring feeding styles in an urban Latina WIC population. AB - OBJECTIVE: Controlling feeding styles in which parents regulate feeding without responding to child cues have been associated with poor self-regulation of feeding and increased weight, but have not been well studied in infancy. We sought to assess maternal perception of infant feeding cues and pressuring feeding styles in an urban Latina Special Supplemental Nutrition Program for Women, Infants, and Children (WIC) population. METHODS: Secondary analysis of a larger study of Latina mothers participating in New York City WIC programs. We examined maternal perception of infant feeding cues and pressuring feeding style. Using logistic regression, we assessed: 1) characteristics associated with perceptions of cues and pressuring to feed, including sociodemographics, breastfeeding, and maternal body mass index; and 2) whether perceptions of cues were associated with pressuring feeding style. RESULTS: We surveyed 368 mothers (84% response rate). Most mothers perceived that babies sense their own satiety. However, 72% believed that infant crying must indicate hunger. Fifty-three percent believed that mothers should always make babies finish the bottle ("pressure to feed"). Pressuring feeding style was associated with foreign maternal country of birth (adjusted odds ratio [AOR] 3.05; 95% confidence interval [CI], 1.66-5.60) and less than a high school education (AOR 1.81; 95% CI, 1.12-2.91). Two perceptions of feeding cues were related to pressuring feeding style: belief that infant crying must indicate hunger (AOR 2.59; 95% CI, 1.52-4.42) and infant hand sucking implies hunger (AOR 1.83; 95% CI, 1.10-3.03). CONCLUSIONS: Maternal characteristics influence perception of infant hunger and satiety. Interpretation of feeding cues is associated with pressuring feeding style. Improving responsiveness to infant cues should be a component of early childhood obesity prevention. PMID- 20004634 TI - Breast cancer risk and common single nucleotide polymorphisms in homologous recombination DNA repair pathway genes XRCC2, XRCC3, NBS1 and RAD51. AB - The possible role for DNA repair deficiencies in cancer development, namely in breast cancer has been the subject of increasing interest since it has been reported that breast cancer patients might be deficient in the repair of DNA damage. Exposure to ionizing radiation has been pointed out as a risk factor for breast cancer, and the type of DNA lesions induced by this carcinogen can be repaired by homologous recombination DNA repair (HRR) pathway. To evaluate the potential modifying role of some single nucleotide polymorphisms (SNP) in HRR involved genes on the individual susceptibility to breast cancer we carried out a hospital based case-control study in a Caucasian Portuguese population (289 histological confirmed breast cancer patients and 548 control individuals). We genotyped 4 SNPs in 4 different HRR pathway genes, XRCC2 (Ex3+442G>A, R188H, rs3218536), XRCC3 (Ex8-5C>T, T241M, rs861539), NBS1 (Ex5-32C>G, E185Q, rs1805794) and RAD51 5'UTR (Ex1-59G>T, rs1801321), tagging 41 SNPs in these genes. The frequency of the different polymorphisms in the Portuguese control population is similar to the ones reported for other Caucasian populations, and the deviation of the Hardy-Weinberg equilibrium was only observed for the XRCC2 (Ex3+442G>A, R188H, rs3218536) polymorphism in the control population. The results obtained, after logistic regression analysis, did not reveal a major role of these polymorphisms on breast cancer susceptibility. However, when the population was stratified according to breast feeding (women that breast fed and women that never breast fed) it is observed, in women that never breast fed, that the heterozygous individuals for the XRCC2 (Ex3+442G>A, R188H, rs3218536) polymorphism have a decreased risk for breast cancer [adjusted OR=0.45; 95% CI=0.22-0.92] (P=0.03). Additionally, after stratification according to menopausal status, our results suggest that post-menopausal women carrying at least one variant allele for the XRCC3 (Ex8-5C>T, T241M, rs861539) polymorphism have a lower risk for breast cancer [adjusted OR=0.67; 95% CI, 0.47-0.94] (P=0.03). Most of the studies suggest that breastfeeding may be responsible for 2/3 of the estimate reduction of breast cancer. The longer the duration of breastfeeding the lower the potential risk associated with breast cancer. Therefore, in our study the potential protective role of the variant allele of XRCC2 (Ex3+442G>A, R188H, rs3218536), in never breast fed women, might be related with a more efficient DNA repair activity. PMID- 20004635 TI - Epidemiological, antigenic and genetic characteristics of seasonal influenza A(H1N1), A(H3N2) and B influenza viruses: basis for the WHO recommendation on the composition of influenza vaccines for use in the 2009-2010 northern hemisphere season. AB - Influenza vaccines form an important component of the global response against infections and subsequent illness caused in man by influenza viruses. Twice a year, in February and September, the World Health Organisation through its Global Influenza Surveillance Network (GISN), recommends appropriate influenza viruses to be included in the seasonal influenza vaccine for the upcoming Northern and Southern Hemisphere winters. This recommendation is based on the latest data generated from many sources and the availability of viruses that are suitable for vaccine manufacture. This article gives a summary of the data and background to the recommendations for the 2009-2010 Northern Hemisphere influenza vaccine formulation. PMID- 20004637 TI - Exonuclease III action on microarrays: observation of DNA degradation by fluorescence correlation spectroscopy. AB - DNA with all cytosines, thymines, or all pyrimidines of one strand substituted by fluorescently labeled analogs shows diminished solubility in aqueous media and a strong tendency to aggregation that hampers enzymatic downstream processing. In this study, immobilization of fully fluorescently labeled DNA on microarrays was shown to resolve the named problems and to enable successive DNA degradation by exonuclease III. Fluorescence correlation spectroscopy and single-molecule counting for monitoring the course of DNA hydrolysis in real time revealed the virtually processive degradation of labeled DNA that occurred at an average rate of approximately 4 nt/s. PMID- 20004636 TI - Cartilage shear dynamics during tibio-femoral articulation: effect of acute joint injury and tribosupplementation on synovial fluid lubrication. AB - OBJECTIVE: To determine the effects of acute injury and tribosupplementation by hyaluronan (HA) on synovial fluid (SF) modulation of cartilage shear during tibio femoral articulation. METHODS: Human osteochondral blocks from the lateral femoral condyle (LFC) and tibial plateau (LTP) were apposed, compressed 13%, and subjected to sliding under video microscopy. Tests were conducted with equine SF from normal joints (NL-SF), SF from acutely injured joints (AI-SF), and AI-SF to which HA was added (AI-SF+HA). Local and overall shear strain (E(xz)) and the lateral displacement (Deltax) at which E(xz) reached 50% of peak values (Deltax(1/2)) were determined. RESULTS: During articulation, LFC and LTP cartilage E(xz) increased with Deltax and peaked when surfaces slid, with peak E(xz) being maintained during sliding. With AI-SF as lubricant, surface and overall Deltax(1/2) were approximately 40% and approximately 20% higher, respectively, than values with NL-SF and AI-SF+HA as lubricant. Also, peak E(xz) was markedly higher with AI-SF as lubricant than with NL-SF as lubricant, both near the surface (approximately 80%) and overall (50-200%). Following HA supplementation to AI-SF, E(xz) was reduced from values with AI-SF alone by 30 50% near the surface and 20-30% overall. Magnitudes of surface and overall E(xz) were markedly (approximately 50 to 80%) higher in LTP cartilage than LFC cartilage for all lubricants. CONCLUSION: Acute injury impairs SF function, elevating cartilage E(xz) markedly during tibio-femoral articulation; such elevated E(xz) may contribute to post-injury associated cartilage degeneration. Since HA partially restores the function of AI-SF, as indicated by E(xz), tribosupplements may be beneficial in modulating normal cartilage homeostasis. PMID- 20004638 TI - Low-temperature electron transfer suggests two types of Q(A) in intact photosystem II. AB - The correlation between the reduction of Q(A) and the oxidation of Tyr(Z) or Car/Chl(Z)/Cyt(b559) in spinach PSII enriched membranes induced by visible light at 10 K is studied by using electron paramagnetic resonance spectroscopy. Similar g=1.95-1.86 Q(A)(-*)EPR signals are observed in both Mn-depleted and intact samples, and both signals are long lived at low temperatures. The presence of PPBQ significantly diminished the light induced EPR signals from Q(A)(-*), Car(+*)/Chl(+*) and oxidized Cyt(b559), while enhancing the amplitude of the S(1)Tyr(Z)* EPR signal in the intact PSII sample. The quantification and stability of the g=1.95-1.86 EPR signal and signals arising from the oxidized Tyr(Z) and the side-path electron donors, respectively, indicate that the EPR detectable g=1.95-1.86 Q(A)(-*) signal is only correlated to reaction centers undergoing oxidation of the side-path electron donors (Car/Chl(Z)/Cyt(b559)), but not of Tyr(Z). These results imply that two types of Q(A)(-*) probably exist in the intact PSII sample. The structural difference and possible function of the two types of Q(A) are discussed. PMID- 20004639 TI - Infrared reflection-absorption spectroscopy: principles and applications to lipid protein interaction in Langmuir films. AB - Infrared reflection-absorption spectroscopy (IRRAS) of lipid/protein monolayer films in situ at the air/water interface provides unique molecular structure and orientation information from the film constituents. The technique is thus well suited for studies of lipid/protein interaction in a physiologically relevant environment. Initially, the nature of the IRRAS experiment is described and the molecular structure information that may be obtained is recapitulated. Subsequently, several types of applications, including the determination of lipid chain conformation and tilt as well as elucidation of protein secondary structure are reviewed. The current article attempts to provide the reader with an understanding of the current capabilities of IRRAS instrumentation and the type of results that have been achieved to date from IRRAS studies of lipids, proteins, and lipid/protein films of progressively increasing complexity. Finally, possible extensions of the technology are briefly considered. PMID- 20004640 TI - Oligo-(R)-3-hydroxybutyrate modification of sorting signal enables pore formation by Escherichia coli OmpA. AB - The outer membrane protein A (OmpA) of Escherichia coli is a well-known model for protein targeting and protein folding. Wild-type OmpA, isolated either from cytoplasmic inclusion bodies or from outer membranes, forms narrow pores of approximately 80 pS in planar lipid bilayers at room temperature. The pores are well structured with narrow conductance range when OmpA is isolated using lithium dodecyl sulfate (LDS) or RapiGest surfactant but display irregular conductance when OmpA is isolated with urea or guanidine hydrochloride. Previous studies have shown that serine residues S163 and S167 of the sorting signal of OmpA (residues 163-169), i.e., the essential sequence for outer membrane incorporation, are covalently modified by oligomers of (R)-3-hydroxybutyrate (cOHB). Here we find that single-mutants S163 and S167 of OmpA, which still contain cOHB on one serine of the sorting signal, form narrow pores in planar lipid bilayers at room temperature with lower and more irregular conductance than wild-type OmpA, whereas double mutants S163:S167 and S163:V166 of OmpA, with no cOHB on the sorting signal, are unable to form stable pores in planar lipid bilayers. Our results indicate that modification of serines in the sorting signal of OmpA by cOHB in the cytoplasm enables OmpA to incorporate into lipid bilayers at room temperature as a narrow pore. They further suggest that cOHB modification may be an important factor in protein targeting and protein folding. PMID- 20004641 TI - P-glycoprotein substrate transport assessed by comparing cellular and vesicular ATPase activity. AB - We compared the P-glycoprotein ATPase activity in inside-out plasma membrane vesicles and living NIH-MDR1-G185 cells with the aim to detect substrate transport. To this purpose we used six substrates which differ significantly in their passive influx through the plasma membrane. In cells, the cytosolic membrane leaflet harboring the substrate binding site of P-glycoprotein has to be approached by passive diffusion through the lipid membrane, whereas in inside-out plasma membrane vesicles, it is accessible directly from the aqueous phase. Compounds exhibiting fast passive influx compared to active efflux by P glycoprotein induced similar ATPase activity profiles in cells and inside-out plasma membrane vesicles, because their concentrations in the cytosolic leaflets were similar. Compounds exhibiting similar influx as efflux induced in contrast different ATPase activity profiles in cells and inside-out vesicles. Their concentration was significantly lower in the cytosolic leaflet of cells than in the cytosolic leaflet of inside-out membrane vesicles, indicating that P glycoprotein could cope with passive influx. P-glycoprotein thus transported all compounds at a rate proportional to ATP hydrolysis (i.e. all compounds were substrates). However, it prevented substrate entry into the cytosol only if passive influx of substrates across the lipid bilayer was in a similar range as active efflux. PMID- 20004642 TI - Molecular shape and binding force of Mycoplasma mobile's leg protein Gli349 revealed by an AFM study. AB - Recent studies of the gliding bacteria Mycoplasma mobile have identified a family of proteins called the Gli family which was considered to be involved in this novel and yet fairly unknown motility system. The 349kDa protein called Gli349 was successfully isolated and purified from the bacteria, and electron microscopy imaging and antibody experiments led to the hypothesis that it acts as the "leg" of M. mobile, responsible for attachment to the substrate as well as for gliding motility. However, more precise evidence of the molecular shape and function of this protein was required to asses this theory any further. In this study, an atomic force microscope (AFM) was used both as an imaging and a force measurement device to provide new information about Gli349 and its role in gliding motility. AFM images of the protein were obtained revealing a complex structure with both rigid and flexible parts, consistent with previous electron micrographs of the protein. Single-molecular force spectroscopy experiments were also performed, revealing that Gli349 is able to specifically bind to sialyllactose molecules and withstand unbinding forces around 70pN. These findings strongly support the idea that Gli349 is the "leg" protein of M. mobile, responsible for binding and also most probably force generation during gliding motility. PMID- 20004643 TI - Anti-tumor effect of small interfering RNA targeting the androgen receptor in human androgen-independent prostate cancer cells. AB - Early phase prostate cancer is usually androgen-dependent, with the androgen/androgen receptor (AR) signaling pathway playing a central role. At this stage, the cancer responds well to androgen ablation therapy, but prostate cancers eventually acquire androgen independence and more aggressive phenotypes. Several studies, however, have shown that the majority of tumors still express functional AR, which is often amplified and mutated. To determine if the AR is a plausible therapeutic target, we investigated the anti-tumor effect of small interfering RNAs targeting the AR (siAR) in the human prostate cancer cells, LNCaP and 22Rv1, which express mutated AR. In both types of cells, transfection of siAR suppressed mutated AR expression and significantly reduced cell growth. Furthermore, atelocollagen-mediated systemic siAR administration markedly inhibited the growth of 22Rv1 cells subcutaneously xenografted in castrated nude mice. These results suggest that the AR is still a key therapeutic target even in androgen-independent prostate cancer (AIPC). Silencing of AR expression in AIPC opens promising therapeutic perspectives. PMID- 20004644 TI - Novel sinomenine derivative 1032 improves immune suppression in experimental autoimmune encephalomyelitis. AB - Sinomenine (SIN) is an alkaloid isolated from the Chinese medicinal plant Sinomenium acutum. It is widely used as an immunosuppressive drug for treating autoimmune diseases. Due to its poor efficiency, the large-dose treatment presents some side effects and limits its further applications. In this study, we used chemical modification to improve the therapeutic effect of SIN in vitro and in vivo. A new derivative of sinomenine, named 1032, demonstrates significantly improved immunosuppressive activity over that of its parent natural compound (SIN). In an experimental autoimmune encephalomyelitis (EAE) model, 1032 significantly reduced encephalitogenic T cell responses and induced amelioration of EAE, which outcome was related to its selective inhibitory effect on the production of IL-17. By contrast, SIN treatment only led to a moderate alleviation of EAE severity and the expression level of IL-17 was not significantly reduced. Furthermore, 1032 exhibited suppression of Th17, but not Treg, cell differentiation, a result probably related to its inhibitory effect on IkappaB-alpha degradation as well as on IL-6 and TNF-alpha secretion in BMDCs. We speculate that 1032 as a novel anti-inflammatory agent may target DC to block IL 6 production, which in turn would terminate Th17 cell development. Thus, SIN derivative 1032 presents considerable potential in new drug development for treating autoimmune and inflammatory disease. PMID- 20004645 TI - Pili of oral Streptococcus sanguinis bind to fibronectin and contribute to cell adhesion. AB - Streptococcus sanguinis is a predominant bacterium in the human oral cavity and occasionally causes infective endocarditis. We identified a unique cell surface polymeric structure named pili in this species and investigated its functions in regard to its potential virulence. Pili of S. sanguinis strain SK36 were shown to be composed of three distinctive pilus proteins (PilA, PilB, and PilC), and a pili-deficient mutant demonstrated reduced bacterial adherence to HeLa and human oral epithelial cells. PilC showed a binding ability to fibronectin, suggesting that pili are involved in colonization by this species. In addition, ATCC10556, a standard S. sanguinis strain, was unable to produce pili due to defective pilus genes, which indicates a diversity of pilus expression among various S. sanguinis strains. PMID- 20004646 TI - Tumor necrosis factor-alpha increases alkaline phosphatase expression in vascular smooth muscle cells via MSX2 induction. AB - Vascular calcification is implicated in many diseases including atherosclerosis and diabetes. Tumor necrosis factor-alpha (TNF-alpha) has been shown to promote vascular calcification both in vitro and in vivo. However, the molecular mechanism of TNF-alpha-mediated vascular calcification has not yet been fully defined. Therefore, in this study, we aimed to investigate whether MSX2 acts as a crucial regulator in TNF-alpha-induced vascular calcification and to define the regulatory mechanism of MSX2 induction in human vascular smooth muscle cells (VSMCs). TNF-alpha increased the expression of osteogenic marker genes including RUNX2, osterix, alkaline phosphatase (ALP), and bone sialoprotein, and it also promoted matrix mineralization in VSMCs. In addition, TNF-alpha enhanced MSX2 expression in a dose- and time-dependent manner. MSX2 over-expression alone induced ALP expression, whereas knockdown of MSX2 with small interfering RNA completely blocked TNF-alpha-induced ALP expression. New protein synthesis was dispensable for MSX2 induction by TNF-alpha, and the inhibition of NF-kappaB by BAY-11-7082 or by dominant negative IkappaBalpha abolished the TNF-alpha-directed induction of MSX2 expression. However, inhibition of NADPH oxidase did not affect MSX2 expression. In conclusion, our study suggests that TNF-alpha directly induces MSX2 expression through the NF-kappaB pathway, which in turn induces expression of ALP, a key molecule in mineralization, in VSMCs. PMID- 20004647 TI - Pharmacological targeting reveals distinct roles for CXCR2/CXCR1 and CCR2 in a mouse model of arthritis. AB - Neutrophils and monocytes are abundantly represented in the synovial fluid and tissue in rheumatoid arthritis patients. We therefore explored the effects of small molecule chemokine receptor antagonists to block migration of these cells in anti-collagen antibody-induced arthritis. Targeting neutrophil migration with the CXCR2/CXCR1 antagonist SCH563705 led to a dose-dependent decrease in clinical disease scores and paw thickness measurements and clearly reduced inflammation and bone and cartilage degradation based on histopathology and paw cytokine analyses. In contrast, targeting monocyte migration with the CCR2 antagonist MK0812 had no effect on arthritis disease severity. The pharmacodynamic activities of both SCH563705 and MK0812 were verified by assessing their effects on the peripheral blood monocyte and neutrophil populations. SCH563705 selectively reduced the peripheral blood neutrophil frequency, and caused an elevation in the CXCR2 ligand CXCL1. MK0812 selectively reduced the peripheral blood monocyte frequency, and caused an elevation in the CCR2 ligand CCL2. The much greater impact of CXCR2/CXCR1 antagonism relative to CCR2 antagonism in this model of arthritis highlights the therapeutic potential for targeting CXCR2/CXCR1 in human arthritides. PMID- 20004648 TI - An in vitro model that can distinguish between effects on angiogenesis and on established vasculature: actions of TNP-470, marimastat and the tubulin-binding agent Ang-510. AB - In anti-cancer therapy, current investigations explore the possibility of two different strategies to target tumor vasculature; one aims at interfering with angiogenesis, the process involving the outgrowth of new blood vessels from pre existing vessels, while the other directs at affecting the already established tumor vasculature. However, the majority of in vitro model systems currently available examine the process of angiogenesis, while the current focus in anti vascular therapies moves towards exploring the benefit of targeting established vasculature as well. This urges the need for in vitro systems that are able to differentiate between the effects of compounds on angiogenesis as well as on established vasculature. To achieve this, we developed an in vitro model in which effects of compounds on different vascular targets can be studied specifically. Using this model, we examined the actions of the fumagillin derivate TNP-470, the MMP-inhibitor marimastat and the recently developed tubulin-binding agent Ang 510. We show that TNP-470 and marimastat solely inhibited angiogenesis, whereas Ang-510 potently inhibited angiogenesis and caused massive disruption of newly established vasculature. We show that the use of this in vitro model allows for specific and efficient screening of the effects of compounds on different vascular targets, which may facilitate the identification of agents with potential clinical benefit. The indicated differences in the mode of action between marimastat, TNP-470 and Ang-510 to target vasculature are illustrative for this approach. PMID- 20004649 TI - Transplantation of placenta-derived mesenchymal stromal cells upon experimental stroke in rats. AB - The beneficial effects of bone marrow-derived mesenchymal stromal cell (MSC) administration following experimental stroke have already been described. Despite several promising characteristics, placenta-derived MSC have not been used in models of focal ischemia. The aim of the current study is to investigate the impact of intravenously transplanted placenta-derived MSC on post-stroke recovery. Permanent occlusion of the middle cerebral artery was induced in spontaneously hypertensive rats. MSC were obtained from the human maternal or fetal placenta and intravenously administered after 24 h (single transplantation) or after 8 h and 24 h (dual transplantation). Sensorimotor deficits were quantified for 60 days using the beam walk test and the modified Neurological Severity Score system. Infarct volume was determined in vivo by means of magnetic resonance imaging on days 1, 8, 29 and 60. Astroglial reactivity was semiquantitatively ascertained within a small and a broad region adjacent to the lesion border. The double infusion of placental MSC was superior to single transplantation in the functional tests. However, a significant difference to the control group in all outcome parameters was observed only for maternally derived MSC. These findings suggest that placental tissue constitutes a promising source for experimental stroke therapies. PMID- 20004650 TI - Alzheimer-type tau pathology in advanced aged nonhuman primate brains harboring substantial amyloid deposition. AB - We elucidated how Alzheimer-type pathologies of amyloid beta-protein (Abeta) and tau spatiotemporally emerge in brains of nontransgenic nonhuman primate, cynomolgus monkey, in the present study. To examine the accumulation of deposited Abeta, phosphorylated tau accumulation, intracellular tau accumulation, and neurofibrillary tangle formation, the brains, mainly temporal cortex and hippocampus, of 34 cynomolgus monkeys aged 6 to 36 years were studied by biochemical and histochemical analyses. Biochemically, first, the accumulation of insoluble Abeta was detected in the neocortical (temporal and frontal) and hippocampal regions of animals as young as mid-20s and their levels were extremely high in those of advanced age. The accumulation of phosphorylated tau in the same regions occurred before the age of 20 with poor correlation to the levels of insoluble Abeta. Histologically, intraneuronal and intraoligodendroglial tau accumulation was observed in temporal cortex and hippocampus of animals before the age of 20. In an advanced aged 36-year-old individual, argyrophilic tangles and tau-accumulated dystrophic neurites were markedly observed in the medial temporal area contiguous to limbic structures. Notably, these tau pathologies also emerged, to a lesser extent, in the temporal cortices of advanced aged animals harboring significant amounts of insoluble Abeta. These results suggest that the cynomolgus monkey can be used to elucidate the age-dependent sequence of Abeta and tau pathologies. PMID- 20004651 TI - Neurocalcin protein labeling reveals a dimorphism within the developing zebra finch brain. AB - The exact mechanism(s) responsible for sexual differentiation of the zebra finch song system remains unknown but likely involve a combination of hormonal and genetic factors. One product that may play a role is the calcium binding protein, neurocalcin. A previous study indicated that neurocalcin mRNA was widely distributed throughout the zebra finch telencephalon, overlapping with song control regions. Because it was not clear how much of that expression was functionally relevant, we labeled tissue with a specific antibody. At the three age ranges that were examined, there were region-specific fluctuations in the number of neurocalcin immunoreactive cells. During post-hatching developmental ages (P) 18-23, quantitative analyses indicated that females had significantly more neurons containing neurocalcin protein in HVC and RA than males. This difference was not detected at P10 or in adults. Labeling in LMAN did not reveal a sex difference at any of the ages. In other areas of the brain, semi quantitative analyses of the relative number of cells immunopositive for neurocalcin varied across specific regions, but with the possible exception of the lateral striatum, no obvious sex differences were apparent. Taken together, these data are consistent with the idea that neurocalcin may be important for regulating sexual dimorphisms within the neural song system at a specific developmental period. In addition, because of variations in events related to post-transcriptional modification, a thorough study of a gene's role in development and/or behavior should include investigations of its transcription as well as translation. PMID- 20004652 TI - Autophagy impairment inhibits differentiation of glioma stem/progenitor cells. AB - Despite of similarities between glioma stem/progenitor cells (GSPCs) and neural stem/progenitor cells (NSPCs), inhibition of differentiation is a distinct characteristic of GSPCs. In this study, we investigated the effects of autophagy impairment on inhibition of differentiation of GSPC, and its molecular mechanism. GSPCs were kept by our laboratory; NSPCs were isolated from human fetal brain tissue. We found that the autophagic activity in GSPCs was significantly lower than that in NSPCs. However, the autophagic activity markedly increased after GSPCs were induced to differentiate by fetal calf serum (FCS). The autophagy inhibitors 3-methyladenine and Bafilomycin A1 (BFA) inhibited the FSC-induced differentiation of GSPCs. And autophagy activator Rapamycin could promote differentiation of GSPCs. In order to disclose whether the loss of PTEN in GSPC is related to the deficiency of autophagic activity in GSPCs (for PTEN being lost in the GSPCs studied by us), we introduced the wild type gene of PTEN into GSPCs, and found that the autophagic activity was restored significantly after the gene transduction. The low autophagic activity in GSPCs leads to the inhibition of differentiation of GSPCs, and the loss of PTEN in GSPCs probably is an underlying mechanism for the low autophagic activity in GSPCs. These results suggest that bust autophagic activity target at PTEN might be a potential therapy target for glioma therapy. PMID- 20004653 TI - SCA17 repeat expansion: mildly expanded CAG/CAA repeat alleles in neurological disorders and the functional implications. AB - BACKGROUND: Spinocerebellar ataxia type 17 (SCA17) involves the expression of a CAG/CAA expansion mutation in the gene encoding TATA-box binding protein (TBP), a general transcription initiation factor. The spectrum of SCA17 clinical presentation is broad. METHODS: We screened for triplet expansion in the TBP gene in Taiwanese Parkinson's disease (PD), Alzheimer's disease (AD) and atypical parkinsonism and investigated the functional implication of expanded alleles using lymphoblastoid cells as a model. RESULTS: A total of 6 mildly expanded alleles (44-46) were identified in patients group. The frequency of the individuals carrying expanded alleles in PD (3/602 [0.5%]), AD (2/245 [0.8%]) and atypical parkinsonism (1/44 [2.3%]) is not significant as compared to that in the control subjects (0/644 [0.0%]). In lymphoblastoid cells, HSPA5, HSPA8 and HSPB1 expression levels in cells with expanded TBP were significantly lower than that of the control cells. Although not significantly, the levels of PARK7 protein isoforms 6.1 and 6.4 are notably increased in SCA17 lymphoblastoid cells. Treatment of TBH (tert-butyl hydroperoxide) significantly increases cell death in the cells with mildly expanded TBP. CONCLUSIONS: Our findings expand the spectrum of SCA17 phenotype and may contribute to our understanding of the disease. PMID- 20004654 TI - Ion Selective Electrodes (ISEs) and interferences--a review. AB - Ion Selective Electrodes (ISEs) are used to measure some of the most critical analytes on clinical laboratory and point-of-care analysers. These analytes which include Na(+), K(+), Cl(-), Ca(2+), Mg(2+) and Li(+) are used for rapid patient care decisions. Although the electrodes are very selective, they are not free of interferences. It is important for laboratories to have an understanding of the type and extent of interferences in order to avoid incorrect clinical decisions and treatment. PMID- 20004655 TI - Significant impact of pace of eating on serum ghrelin and glucose levels. AB - OBJECTIVES: To study the effect of normal versus slow eating on serum ghrelin, glucose, insulin, and C-peptide levels in healthy subjects from Riyadh, Saudi Arabia. DESIGN AND METHODS: The specified breakfast meal was served on two randomized occasions to 24 healthy volunteers to eat with a normal pace or at a slow rate. Venous blood samples were collected at 7 time points for biochemical analysis. RESULTS: The slow ingestion of meals resulted in a significant increase in blood glucose and ghrelin levels as compared to normal pace of eating. CONCLUSIONS: Normal eating speed appears to be beneficial for maintaining the optimal blood glucose and ghrelin levels. PMID- 20004656 TI - Meig1 deficiency causes a severe defect in mouse spermatogenesis. AB - Meig1 is a mouse gene, abundantly expressed in the testis. It encodes two alternative transcripts that are expressed differentially in the somatic and germinal compartments of the testis. These transcripts share the same coding region but differ in their 5' un-translated regions, due to alternative promoters. Here we show that MEIG1 is a highly conserved short metazoan protein with a conserved core of 81 residues. It is present from chordates to radial symmetry animals, with an intriguing absence in insects and nematodes. It is also present in two earlier diverging protist lineages. To elucidate the role of MEIG1 during gamete production we established a knockout mouse line by eliminating the common coding region. Our results identified Meig1 as a critical spermatogenic gene, whose absence results in complete male infertility. Seminiferous tubules in Meig1-null males contained all early stages of spermatogenesis, up to elongating spermatids, but mature elongated spermatids were absent. Accordingly, the caudal epididymis was apparently missing spermatozoa, and the very few spermatozoa-like cells that were obtained were immotile and exhibited a wide range of severe morphological abnormalities. These results point at late spermiogenesis as the differentiative stage at which MEIG1's function is crucial. Nevertheless, delayed kinetics of earlier meiotic stages together with increased apoptosis of meiotic spermatocytes and haploid round spermadids in Meig1 knockout males, suggest involvement of MEIG1 in meiotic stages as well. PMID- 20004657 TI - Effect of pioglitazone on insulin resistance in fructose-drinking rats correlates with AGEs/RAGE inhibition and block of NADPH oxidase and NF kappa B activation. AB - Insulin resistance, which may influence Alzheimer's disease, is associated with an increase in circulating advanced glycosylation end products (AGEs) and the increased expression of the receptor for AGEs (RAGE). Inhibition of AGE/RAGE system has been shown to attenuate neuronal damage. Specific ligands for peroxisome proliferator-activated receptor gamma (PPARgamma), which have proven effective in the animal models of Alzheimer's disease and other neuroinflammatory diseases, have been shown to decrease RAGE expression. Here we investigated the effect of PPARgamma agonist, pioglitazone, on cognition function and AGE/RAGE system in a rodent model of insulin resistance, the fructose-drinking rats. Six week-old male Wistar rats were fed a standard commercial diet and water without (control) or with 10% fructose for 16weeks. The animals were randomly divided into 4 groups (n=10): non-treated and water-drinking rats (control group); pioglitazone-treated and water-drinking (control treatment group); non-treated and fructose-drinking rats (fructose group) and pioglitazone-treated and fructose drinking rats (fructose treatment group). Pioglitazone was given at the dose of 10mg/kgd by gavage for the last 12weeks of the 16-week period. The results showed that pioglitazone treatment reduced the escape latency in Morris water maze test, decreased AGE/RAGE expression in the cerebral cortex of fructose-drinking rats. Furthermore, we found that the expression of p47phox component of NAPDH oxidase, phosphorylated nuclear factor NF-kappaB p65, tumor necrosis factor alpha and interleukin-1 beta was significantly increased in the cerebral cortex of fructose drinking rats (P<0.001). These effects were reversed by pioglitazone treatment (P<0.01 or 0.001). Taken together, these findings suggest that the activation of AGEs-RAGE system contribute to the brain damage of insulin resistance. Pioglitazone administration can improve cognition function probably related to its effect of decreasing the activation of AGEs-RAGE system, which correlates with block of NAPDH oxidase and NF-kappaB activation in this rodent model of insulin resistance. PMID- 20004658 TI - Blockade of dopamine, but not noradrenaline, transporters produces hyperthermia in rats that lack serotonin transporters. AB - To investigate whether life-long disturbed serotonin neurotransmission may result in adaptive changes of dopaminergic and noradrenergic systems, effects of drugs on stress-induced hyperthermia were studied in serotonin transporter knockout rats. The noradrenalin transporter blocker atomoxetine was more effective in reducing stress-induced hyperthermia, induced by an injection, in serotonin transporter (SERT) knockout (SERT(-/-)) rats compared to SERT(+/+) rats. The dopamine transporter blocker GBR12909 increased the core body temperature in SERT(-/-) rats, and had no effect on the SERT(+/+) rats. Finally, the noradrenalin transporter together with dopamine transporter blocker bupropion was more effective in decreasing the stress of an injection in SERT(-/-) rats than in SERT(+/+) rats. These data suggest that the sensitivity of dopamine and noradrenalin receptors is changed in serotonin transporter knockout rats. The lack of the serotonin transporter in SERT(-/-) rats might reflect humans with a life-long disturbed serotonin system, making this rat a good model to study possible changes in dopaminergic and noradrenergic systems in psychiatric disorders. PMID- 20004659 TI - Cortactin is implicated in murine zygotic development. AB - Cortactin is a cortex-enriched protein implicated in Arp2/3 complex-mediated actin polymerization. However, the physiological role of cortactin remains unknown. We have generated a mouse strain in which the allele of murine cortactin was disrupted by a gene trapping vector. The resulting heterozygous mice developed normally and were fertile, but embryonic fibroblasts derived from heterozygous animals displayed partial impairment in PDGF-induced membrane ruffling. No homozygous offspring or early embryos even at the two-cell stage were detected. Analysis of oocytes revealed a gradual decrease in the detection of homozygous zygotes after fertilization. In normal oocytes arrested at meiotic metaphase II (MII), cortactin immunoreactivity was detected in an apical layer that overlies the maternal chromosome and overlaps with a polarized cortex enriched with actin. The formation of the polarized cortactin layer was diminished upon treatment with latrunculin B, an actin polymerization inhibitor. After resumption of meiosis II, the majority of cortactin protein was accumulated into the second polar body. Microinjection of MII-arrested eggs with either cortactin antibody or RNA encoding a cortactin mutant deficient in Arp2/3 complex binding disrupted the integrity of the actin cap and inhibited emission of the second polar body triggered by parthenogenesis. Our data suggest that cortactin plays an important role in the mechanics of asymmetric division in oocytes. PMID- 20004660 TI - Introduction to special issue: Challenges and opportunities for regeneration in the peripheral nervous system. AB - Regeneration in the peripheral nervous system offers unique opportunities and challenges to medicine. Compared to the central nervous system, peripheral axons can and do regenerate resulting in functional recovery, especially if the distance to target is short as in distal limb injuries. However, this regenerative capacity is often incomplete and functional recovery with proximal lesions is limited. Furthermore, regeneration of axons to the appropriate targets remains a challenge with inappropriate reinnervation being an impediment to full recovery. The reviews and selected original research papers in this Special Issue will address some of these challenges and highlight new opportunities for development of effective therapies for nerve regeneration. PMID- 20004662 TI - Mixing pleasures: review of the effects of drugs on sex behavior in humans and animal models. AB - Drugs of abuse act on the brain circuits mediating motivation and reward associated with natural behaviors. There is ample evidence that drugs of abuse impact male and female sexual behavior. First, the current review discusses the effect of drugs of abuse on sexual motivation and performance in male and female humans. In particular, we discuss the effects of commonly abused drugs including psychostimulants, opiates, marijuana/THC, and alcohol. In general, drug use affects sexual motivation, arousal, and performance and is commonly associated with increased sexual risk behaviors. Second, studies on effects of systemic administration of drugs of abuse on sexual behavior in animals are reviewed. These studies analyze the effects on sexual performance and motivation but do not investigate the effects of drugs on risk-taking behavior, creating a disconnect between human and animal studies. For this reason, we discuss two studies that focus on the effects of alcohol and methamphetamine on inhibition of maladaptive sex-seeking behaviors in rodents. Third, this review discusses potential brain areas where drugs of abuse may be exerting their effect on sexual behavior with a focus on the mesolimbic system as the site of action. Finally, we discuss recent studies that have brought to light that sexual experience in turn can affect drug responsiveness, including a sensitized locomotor response to amphetamine in female and male rodents as well as enhanced drug reward in male rats. PMID- 20004661 TI - Farglitazar lacks antifibrotic activity in patients with chronic hepatitis C infection. AB - BACKGROUND & AIMS: Farglitazar (GI262570), an insulin-sensitizing agent, selectively binds and activates peroxisome proliferator-activated receptor gamma (PPARgamma) and inhibits stellate cell activation. We evaluated its antifibrotic effect in patients with chronic hepatitis C that did not respond to standard-of care therapy. METHODS: Patients with fibrosis of Ishak stages 2-4 (n = 265), based on analysis of liver biopsy samples, were randomly assigned to groups given once-daily doses of 0.5 mg farglitazar, 1.0 mg farglitazar, or placebo for 52 weeks; repeat liver biopsy samples were then obtained. The primary end points were changes in levels of alpha-smooth muscle actin (SMA) expression and collagen, based on morphometry and ranked histologic assessments. RESULTS: Two hundred nine patients had paired biopsy specimens adequate for analysis (81.5% with pretreatment Ishak scores of stage 2 or 3). There was no overall difference in SMA (P = .58) or collagen (P = .99) levels at week 52. SMA levels increased by a median of 49% in samples from patients given placebo, 58% in patients given 0.5 mg farglitizar and 52% in patients given 1.0 mg farglitizar, respectively. Collagen increased by 27% in placebo samples and 31% in samples from patients given either dose of farglitizar. There were no significant differences between treatment groups in the ranked assessment of paired biopsy specimens or in the proportion of patients with a change in fibrosis score > or = Ishak stage. CONCLUSIONS: In patients with chronic hepatitis C and moderate fibrosis, 52 weeks of treatment with farglitazar does not affect stellate cell activation or fibrosis (measured by morphometry or comparison of paired biopsy specimens). PMID- 20004663 TI - Cardiac myosin heavy chain isoform exchange alters the phenotype of cTnT-related cardiomyopathies in mouse hearts. AB - Familial hypertrophic cardiomyopathy, FHC, is a clinically heterogeneous, autosomal-dominant disease of the cardiac sarcomere leading to extensive remodeling at both the whole heart and molecular levels. The remodeling patterns are mutation-specific, a finding that extends to the level of single amino acid substitutions at the same peptide residue. Here we utilize two well-characterized transgenic FHC mouse models carrying independent amino acid substitutions in the TM-binding region of cardiac troponin T (cTnT) at residue 92. R92Q and R92L cTnT domains have mutation-specific average peptide conformation and dynamics sufficient to alter thin filament flexibility and cross-bridge formation and R92 mutant myocytes demonstrate mutation-specific temporal molecular remodeling of Ca(2+) kinetics and impaired cardiac contractility and relaxation. To determine if a greater economy of contraction at the crossbridge level would rescue the mechanical defects caused by the R92 cTnT mutations, we replaced the endogenous murine alpha-myosin heavy chain (MyHC) with the beta-MyHC isoform. While beta MyHC replacement rescued the systolic dysfunction in R92Q mice, it failed to rescue the defects in diastolic function common to FHC-associated R92 mutations. Surprisingly, a significant component of the whole heart and molecular contractile improvement in the R92Q mice was due to improvements in Ca(2+) homeostasis including SR uptake, [Ca2+](i) amplitude and phospholamban phosphorylation. Our data demonstrate that while genetically altering the myosin composition of the heart bearing a thin filament FHC mutation is sufficient to improve contractility, diastolic performance is refractory despite improved Ca(2+) kinetics. These data reveal a previously unrecognized role for MyHC isoforms with respect to Ca(2+) homeostasis in the setting of cardiomyopathic remodeling and demonstrate the overall dominance of the thin filament mutation in determining the degree of diastolic impairment at the myofilament level. PMID- 20004664 TI - The molecular basis of the steep force-calcium relation in heart muscle. AB - Contraction of heart muscle is regulated by binding of Ca(2+) ions to troponin in the muscle thin filaments, causing a change in filament structure that allows myosin binding and force generation. The steady-state relationship between force and Ca(2+) concentration in demembranated ventricular trabeculae is well described by the Hill equation, with parameters EC(50), the Ca(2+) concentration that gives half the maximum force, and n(H), the Hill coefficient describing the steepness of the Ca(2)(+) dependence. Although each troponin molecule has a single regulatory Ca(2+) site, n(H) is typically around 3, indicating co operativity in the regulatory mechanism. This review focuses on the molecular basis of this co-operativity, and in particular on the popular hypothesis that force-generating myosin cross-bridges are responsible for the effect. Although cross-bridges can switch on thin filaments at low MgATP concentrations, we argue that the evidence from contracting heart muscle cells shows that this mechanism does not operate in more physiological conditions, and would not play a significant role in the intact heart. Interventions that alter maximum force and EC(50) do not in general produce a significant change in n(H). Complete abolition of force generation by myosin inhibitors does not affect the n(H) values for either Ca(2+) binding to the thin filaments or changes in troponin structure, and both values match that for force generation in the absence of inhibitors. These results provide strong evidence that the co-operative mechanism underlying the high value of n(H) is not due to force-generating cross-bridges but is rather an intrinsic property of the thin filaments. PMID- 20004665 TI - Netrin-1 prevents ischemia/reperfusion-induced myocardial infarction via a DCC/ERK1/2/eNOS s1177/NO/DCC feed-forward mechanism. AB - We have recently shown that a novel endothelial mitogen netrin-1 potently stimulates nitric oxide (NO()) production via a DCC-ERK1/2 dependent mechanism. In view of the well-established cardioprotective role of NO(), the present study investigated whether netrin-1 is cardioprotective via NO(*) signaling in the heart. Netrin-1 receptor DCC was abundantly expressed in the C57BL/6J mouse hearts. Perfusion of heart with netrin-1 (100 ng/mL) using a Langendorff system significantly increased NO(*) production. Under ischemia/reperfusion (I/R), netrin-1 induced a substantial reduction in infarct size (21.8+/-4.9% from 42.5+/ 3.6% in the controls), which was accompanied by an augmented production of NO(*). Pre-perfusion with DCC-antibody, U0126 (MEK1/2 inhibitor), L-NAME or PTIO (NO(*) scavenger) attenuated protective effects of netrin-1 on infarct size and NO(*) production, indicating upstream roles of DCC and ERK1/2 in NO(*) production, as well as an essential role of NO(*) in cardioprotection. Netrin-1 induced reduction in infarct size was significantly attenuated in DCC+/- mice, confirming an intermediate role of DCC. In additional experiments we found netrin-1 increased ERK1/2 and eNOS(s1177) phosphorylation, and DCC protein expression, which was diminished by I/R. Furthermore, netrin-1-induced DCC upregulation was NO(*) and ERK1/2-dependent, implicating a feed-forward mechanism. DAF-AM staining revealed enhanced NO(*) production in both cardiac endothelial cells (ECs) and myocytes. In primarily isolated cardiomyocytes, netrin-1 also increased NO(*) production, DCC abundance and ERK1/2 phosphorylation. Of note, cardiac apoptosis was significantly attenuated by netrin-1, which was reversed by DCC-antibody, U0126, L-NAME or PTIO. In summary, our data clearly demonstrate that netrin-1 potently protects the heart from I/R injury by stimulating NO(*) production from cardiac ECs and myocytes. This potent effect is mediated by a DCC/ERK1/2/eNOS(s1177)/NO(*)/DCC feed-forward mechanism in both cell types. PMID- 20004666 TI - Minicircle performance depending on S/MAR-nuclear matrix interactions. AB - The ideal vector for cell and tissue modification does not depend on integration but rather behaves as an independent functional unit that replicates as an episome. Based on a scaffold/matrix attachment region (S/MAR), we have introduced, in 2006, an approximately 4-kb replicating nonviral minicircle able to exploit the cellular replication machinery in a way reminiscent of ARS vectors. Consisting of only one active transcription unit and the S/MAR, it resists silencing as it is free of prokaryotic vector parts and drug selection markers. The rate of final establishment in the nuclear architecture is moderate but comparable to Epstein-Barr virus-based episomes (<5%). Here, we demonstrate that this parameter can be improved if the host cell chromatin is opened by histone hyperacetylation prior to transfection. It remains unaffected, however, by cell cycle position. Still, this class of episomes revealed intrinsic instability and integration after 5 months of continuous culture. In vivo evolution enabled the effective reduction of S/MAR size from 2 kb to 733 bp (resulting in a minicircle of approximately 3 kb) with largely improved stability and cloning capacity. Investigation of individual clones served to prove persistent and homogenous expression, which is ascribed to stable association with nuclear attachment sites. Optimum expression levels were shown to depend on the authentic usage of a polyadenylation site 3' from the S/MAR as anticipated by the stress-induced duplex destabilization algorithm, which finds increasing use to predict the functional parameters of these systems. PMID- 20004667 TI - The cAMP receptor-like protein CLP is a novel c-di-GMP receptor linking cell-cell signaling to virulence gene expression in Xanthomonas campestris. AB - Cyclic-di-GMP [bis-(3'-5')-cyclic diguanosine monophosphate] controls a wide range of functions in eubacteria, yet little is known about the underlying regulatory mechanisms. In the plant pathogen Xanthomonas campestris, expression of a subset of virulence genes is regulated by c-di-GMP and also by the CAP (catabolite activation protein)-like protein XcCLP, a global regulator in the CRP/FNR superfamily. Here, we report structural and functional insights into the interplay between XcCLP and c-di-GMP in regulation of gene expression. XcCLP bound target promoter DNA with submicromolar affinity in the absence of any ligand. This DNA-binding capability was abrogated by c-di-GMP, which bound to XcCLP with micromolar affinity. The crystal structure of XcCLP showed that the protein adopted an intrinsically active conformation for DNA binding. Alteration of residues of XcCLP implicated in c-di-GMP binding through modeling studies caused a substantial reduction in binding affinity for the nucleotide and rendered DNA binding by these variant proteins insensitive to inhibition by c-di GMP. Together, these findings reveal the structural mechanism behind a novel class of c-di-GMP effector proteins in the CRP/FNR superfamily and indicate that XcCLP regulates bacterial virulence gene expression in a manner negatively controlled by the c-di-GMP concentrations. PMID- 20004668 TI - The outer membrane usher guarantees the formation of functional pili by selectively catalyzing donor-strand exchange between subunits that are adjacent in the mature pilus. AB - Type 1 pili from uropathogenic Escherichia coli are a prototype of adhesive surface organelles assembled and secreted by the conserved chaperone/usher pathway. They are composed of four different homologous protein subunits that need to be assembled in a defined order. In the periplasm, the pilus chaperone FimC donates a beta-strand segment to the subunits to complete their imperfect immunoglobulin-like fold. During subunit assembly, this segment of the chaperone is displaced by an amino-terminal extension of an incoming subunit in a reaction termed donor-strand exchange. To date, the molecular mechanisms underlying the coordinated subunit assembly, in particular the role of the outer membrane usher FimD, are still poorly understood. Here we show that the binding of complexes between FimC and the different pilus subunits to the amino-terminal substrate recognition domain of FimD is an extremely fast process, with association rate constants in the range of 10(7)-10(8) M(-)(1) s(-1) at 20 degrees C. Furthermore, we demonstrate that the ordered assembly of pilus subunits is a consequence of the usher's ability to selectively catalyze the assembly of defined subunit subunit pairs that are adjacent in the mature pilus. The usher therefore coordinates the assembly of pilus subunits at the stage of donor-strand exchange between pairs of subunits and not at the level of the initial binding of chaperone-subunit complexes. PMID- 20004670 TI - Elementary network reconstruction: a framework for the analysis of regulatory networks in biological systems. AB - Complexity of regulatory networks arises from the high degree of interaction between network components such as DNA, RNA, proteins, and metabolites. We have developed a modeling tool, elementary network reconstruction (ENR), to characterize these networks. ENR is a knowledge-driven, steady state, deterministic, quantitative modeling approach based on linear perturbation theory. In ENR we demonstrate a novel means of expressing control mechanisms by way of dimensionless steady state gains relating input and output variables, which are purely in terms of species abundances (extensive variables). As a result of systematic enumeration of network species in nxn matrix, the two properties of linear perturbation are manifested in graphical representations: transitive property is evident in a special L-shape structure, and additive property is evident in multiple L-shape structures arriving at the same matrix cell. Upon imposing mechanistic (lowest-level) gains, network self-assembly through transitive and additive properties results in elucidation of inherent topology and explicit cataloging of higher level gains, which in turn can be used to predict perturbation results. Application of ENR to the regulatory network behind carbon catabolite repression in Escherichia coli is presented. Through incorporation of known molecular mechanisms governing transient and permanent repressions, the ENR model correctly predicts several key features of this regulatory network, including a 50% downshift in intracellular cAMP level upon exposure to glucose. Since functional genomics studies are mainly concerned with redistribution of species abundances in perturbed systems, ENR could be exploited in the system-level analysis of biological systems. PMID- 20004669 TI - Lineage-specific differences in the amino acid substitution process. AB - In Darwinian evolution, mutations occur approximately at random in a gene, turned into amino acid mutations by the genetic code. Some mutations are fixed to become substitutions and some are eliminated from the population. Partitioning pairs of closely related species with complete genome sequences by average population size of each pair, we looked at the substitution matrices generated for these partitions and compared the substitution patterns between species. We estimated a population genetic model that relates the relative fixation probabilities of different types of mutations to the selective pressure and population size. Parameterizations of the average and distribution of selective pressures for different amino acid substitution types in different population size comparisons were generated with a Bayesian framework. We found that partitions in population size as well as in substitution type are required to explain the substitution data. Selection coefficients were found to decrease with increasingly radical amino acid substitution and with increasing effective population size. To further explore the role of underlying processes in amino acid substitution, we analyzed embryophyte (plant) gene families from TAED (The Adaptive Evolution Database), where solved structures for at least one member exist in the Protein Data Bank. Using PAML, we assigned branches to three categories: strong negative selection, moderate negative selection/neutrality, and positive diversifying selection. Focusing on the first and third categories, we identified sites changing along gene family lineages and observed the spatial patterns of substitution. Selective sweeps were expected to create primary sequence clustering under positive diversifying selection. Co-evolution through direct physical interaction was expected to cause tertiary structural clustering. Under both positive and negative selection, the substitution patterns were found to be nonrandom. Under positive diversifying selection, significant independent signals were found for primary and tertiary sequence clustering, suggesting roles for both selective sweeps and direct physical interaction. Under strong negative selection, the signals were not found to be independent. All together, a complex interplay of population genetic and protein thermodynamics forces is suggested. PMID- 20004671 TI - Anomalies in the transcriptional regulatory network of the yeast Saccharomyces cerevisiae. AB - We investigate the structural and dynamical properties of the transcriptional regulatory network of the Yeast Saccharomyces cerevisiae and compare it with two "unbiased" ensembles: one obtained by reshuffling the edges and the other generated by mimicking the transcriptional regulation mechanism within the cell. Both ensembles reproduce the degree distributions (the first-by construction exactly and the second approximately), degree-degree correlations and the k-core structure observed in Yeast. An exceptionally large dynamically relevant core network found in Yeast in comparison with the second ensemble points to a strong bias towards a collective organization which is achieved by subtle modifications in the network's degree distributions. We use a Boolean model of regulatory dynamics with various classes of update functions to represent in vivo regulatory interactions. We find that the Yeast's core network has a qualitatively different behavior, accommodating on average multiple attractors unlike typical members of both reference ensembles which converge to a single dominant attractor. Finally, we investigate the robustness of the networks and find that the stability depends strongly on the used function class. The robustness measure is squeezed into a narrower band around the order-chaos boundary when Boolean inputs are required to be nonredundant on each node. However, the difference between the reference models and the Yeast's core is marginal, suggesting that the dynamically stable network elements are located mostly on the peripherals of the regulatory network. Consistently, the statistically significant three-node motifs in the dynamical core of Yeast turn out to be different from and less stable than those found in the full transcriptional regulatory network. PMID- 20004672 TI - Cycling expression and cooperative operator interaction in the trp operon of Escherichia coli. AB - Oscillatory behaviour in the tryptophan operon of an Escherichia coli mutant strain lacking the enzyme-inhibition regulatory mechanism has been observed by Bliss et al. but not confirmed by others. This behaviour could be important from the standpoint of synthetic biology, whose goals include the engineering of intracellular genetic oscillators. This work is devoted to investigating, from a mathematical modelling point of view, the possibility that the trp operon of the E. coli inhibition-free strain expresses cyclically. For that we extend a previously introduced model for the regulatory pathway of the tryptophan operon in Escherichia coli to account for the observed multiplicity and cooperativity of repressor binding sites. Thereafter we investigate the model dynamics using deterministic numeric solutions, stochastic simulations, and analytic studies. Our results suggest that a quasi-periodic behaviour could be observed in the trp operon expression level of single bacteria. PMID- 20004673 TI - Upregulation of caveolin-1 expression is associated with structural modifications of endothelial cells in diabetic lung. AB - Diabetes and the associated hyperglycemia affect pulmonary physiology and biochemistry inducing endothelial impairment, as the first step in lung vascular dysfunction. Caveolin-1, a characteristic protein of endothelial caveolae, acts as a scaffolding protein involved in signal transduction, cholesterol homeostasis, and vesicular trafficking. To document the effect of hyperglycemia on lung endothelial cells, we designed experiments on streptozotocin-induced diabetes and on double transgenic diabetic mice and investigated (1) the early morphological changes occurring in endothelial cells, (2) the ACE activity and cholesterol content of caveolae-rich membrane microdomains, and (3) the protein and gene expression of caveolin-1. We provide evidence that in diabetic lung, the endothelial cell displays an increased number of caveolae and enlarged surface area and a well-developed synthetic machinery, changes that correlate with an overall augmented ACE activity and cholesterol content and overexpression (gene and protein) of caveolin-1. Targeting the endothelial cell surface molecules modulated by hyperglycemia, such as caveolin-1 and ACE could be an additional therapeutic strategy in diabetes. PMID- 20004674 TI - Alteration of the carbohydrate for deoxyguanosine analogs markedly changes DNA replication fidelity, cell cycle progression and cytotoxicity. AB - Nucleoside analogs are efficacious cancer chemotherapeutics due to their incorporation into tumor cell DNA. However, they exhibit vastly different antitumor efficacies, suggesting that incorporation produces divergent effects on DNA replication. Here we have evaluated the consequences of incorporation on DNA replication and its fidelity for three structurally related deoxyguanosine analogs: ganciclovir (GCV), currently in clinical trials in a suicide gene therapy approach for cancer, D-carbocyclic 2'-deoxyguanosine (CdG) and penciclovir (PCV). GCV and CdG elicited similar cytotoxicity at low concentrations, whereas PCV was 10-100-fold less cytotoxic in human tumor cells. DNA replication fidelity was evaluated using a supF plasmid-based mutation assay. Only GCV induced a dose-dependent increase in mutation frequency, predominantly GC-->TA transversions, which contributed to cytotoxicity and implicated the ether oxygen in mutagenicity. Activation of mismatch repair with hydroxyurea decreased mutations but failed to repair the GC-->TA transversions. GCV slowed S-phase progression and CdG also induced a G2/M block, but both drugs allowed completion of one cell cycle after drug treatment followed by cell death in the second cell cycle. In contrast, PCV induced a lengthy early S-phase block due to profound suppression of DNA synthesis, with cell death in the first cell cycle after drug treatment. These data suggest that GCV and CdG elicit superior cytotoxicity due to their effects in template DNA, whereas strong inhibition of nascent strand synthesis by PCV may protect against cytotoxicity. Nucleoside analogs based on the carbohydrate structures of GCV and CdG is a promising area for antitumor drug development. PMID- 20004675 TI - Modulatory effects of S 38232, a non alpha-7 containing nicotine acetylcholine receptor agonist on network activity in the mouse hippocampus. AB - Extracellular field potentials (fEPSPs) and whole cell patch-clamp recordings were used to test the effect of S 38232, a newly developed potent non-alpha7 nicotinic acetylcholine receptors (nAChR) agonist, on synaptic transmission in hippocampal slices obtained from adult mice. S 38232 increased the amplitude of fEPSPs, evoked in stratum radiatum by Schaffer collateral stimulation. This effect was potentiated by picrotoxin, suggesting that S 38232 exerts at least in part its effect on GABAergic interneurons. The action of S 38232 was mediated by non-alpha7 containing nAChRs since it was prevented by DHbetaE (1muM) but not by alpha-BTX (100nM). A similar potentiating effect on fEPSPs was observed when nicotine (1muM) was applied to hippocampal slices obtained from alpha7 -/- mice in the presence of picrotoxin. The potentiating effect of S 38232 was probably presynaptic in origin since it was associated with a significant reduction in paired-pulse ratio. In addition, in patch clamp experiments, S 38232 enhanced the frequency (but not the amplitude) of spontaneous excitatory and inhibitory postsynaptic currents (sEPSCs, sIPSCs) recorded from CA1 principal cells. Moreover, it enhanced the frequency of miniature IPSCs but not EPSCs, suggesting that it was acting on nAChRs located on presynaptic/pre-terminal regions of GABAergic interneurons. The effect of S 38232 on GABAergic signaling was concentration-dependent with an EC(50) of 43muM. In conclusions, we present evidence that the new nicotine ligand S 38232, by selectively activating non alpha7 nAChRs located on principal cells and GABAergic interneurons, influences network activity and information processing in the hippocampus. PMID- 20004676 TI - Roles of cutaneous versus spinal TRPA1 channels in mechanical hypersensitivity in the diabetic or mustard oil-treated non-diabetic rat. AB - Previous results indicate that intaperitoneal administration of a TRPA1 channel antagonist attenuates diabetic hypersensitivity. We studied whether the antihypersensitivity effect induced by a TRPA1 channel antagonist in diabetic animals is explained by action on the TRPA1 channel in the skin, the spinal cord, or both. For comparison, we determined the contribution of cutaneous and spinal TRPA1 channels to development of hypersensitivity induced by topical administration of mustard oil in healthy controls. Diabetes mellitus was induced by streptozotocin in the rat. Hypersensitivity was assessed by the monofilament- and paw pressure-induced limb withdrawal response. Intrathecal (i.t.) administration of Chembridge-5861528 (CHEM, a TRPA1 channel antagonist) at doses 2.5-5.0 microg/rat markedly attenuated diabetic hypersensitivity, whereas 20 microg of CHEM was needed to produce a weak attenuation of diabetic hypersensitivity with intraplantar (i.pl.) administrations. In controls, i.pl. administration of CHEM (20 microg) produced a weak antihypersensitivity effect at the mustard oil-treated site. I.t. administration of CHEM (10 microg) in controls produced a strong antihypersensitivity effect adjacent to the mustard oil-treated area (site of secondary hyperalgesia), while it failed to influence hypersensitivity at the mustard oil-treated area (site of primary hyperalgesia). A reversible antagonism of the rat TRPA1 channel by CHEM was verified using in vitro patch clamp recordings. The results suggest that while cutaneous TRPA1 channels contribute to mechanical hypersensitivity induced by diabetes or topical mustard oil, spinal TRPA1 channels, probably on central terminals of primary afferent nerve fibers, play an important role in maintenance of mechanical hypersensitivity in these conditions. PMID- 20004678 TI - The sense of body: a multidisciplinary approach to body representation. PMID- 20004677 TI - Selective potentiation of homomeric P2X2 ionotropic ATP receptors by a fast non genomic action of progesterone. AB - P2X receptors are ligand-gated ion channels activated by ATP that are widely expressed in the organism and regulate many physiological functions. We have studied the effect of progesterone (PROG) on native P2X receptors present in rat dorsal root ganglion (DRG) neurons and on recombinant P2X receptors expressed in HEK293 cells or Xenopus laevis oocytes. The effects of PROG were observed and already maximal during the first coapplication with ATP and did not need any preincubation of the cells with PROG, indicating a fast mechanism of action. In DRG neurons, PROG rapidly and reversibly potentiated submaximal but not saturating plateau-like currents evoked by ATP, but had no effect on the currents activated by alpha,beta-methylene ATP, an agonist of homomeric or heteromeric receptors containing P2X1 or P2X3 subunits. In cells expressing homomeric P2X2 receptors, responses to submaximal ATP, were systematically potentiated by PROG in a dose-dependent manner with a threshold between 1 and 10 nM. PROG had no effect on ATP currents carried by homomeric P2X1, P2X3, and P2X4 receptors or by heteromeric P2X1/5 and P2X2/3 receptors. We conclude that PROG selectively potentiates homomeric P2X2 receptors and, in contrast with dehydroepiandrosterone (DHEA), discriminates between homomeric and heteromeric P2X2-containing receptors. This might have important physiological implications since the P2X2 subunit is the most widely distributed P2X subunit in the organism. Moreover, DHEA and PROG might be useful tools to clarify the distribution and the role of native homo- and heteromeric P2X2 receptors. PMID- 20004679 TI - The effects of acute alcohol exposure on the response properties of neurons in visual cortex area 17 of cats. AB - Physiological and behavioral studies have demonstrated that a number of visual functions such as visual acuity, contrast sensitivity, and motion perception can be impaired by acute alcohol exposure. The orientation- and direction-selective responses of cells in primary visual cortex are thought to participate in the perception of form and motion. To investigate how orientation selectivity and direction selectivity of neurons are influenced by acute alcohol exposure in vivo, we used the extracellular single-unit recording technique to examine the response properties of neurons in primary visual cortex (A17) of adult cats. We found that alcohol reduces spontaneous activity, visual evoked unit responses, the signal-to-noise ratio, and orientation selectivity of A17 cells. In addition, small but detectable changes in both the preferred orientation/direction and the bandwidth of the orientation tuning curve of strongly orientation-biased A17 cells were observed after acute alcohol administration. Our findings may provide physiological evidence for some alcohol-related deficits in visual function observed in behavioral studies. PMID- 20004680 TI - Toxicity and toxicokinetics of metformin in rats. AB - Metformin is a first-line drug for the treatment of type 2 diabetes (T2D) and is often prescribed in combination with other drugs to control a patient's blood glucose level and achieve their HbA1c goal. New treatment options for T2D will likely include fixed dose combinations with metformin, which may require preclinical combination toxicology studies. To date, there are few published reports evaluating the toxicity of metformin alone to aid in the design of these studies. Therefore, to understand the toxicity of metformin alone, Crl:CD(SD) rats were administered metformin at 0, 200, 600, 900 or 1200 mg/kg/day by oral gavage for 13 weeks. Administration of > or =900 mg/kg/day resulted in moribundity/mortality and clinical signs of toxicity. Other adverse findings included increased incidence of minimal necrosis with minimal to slight inflammation of the parotid salivary gland for males given 1200 mg/kg/day, body weight loss and clinical signs in rats given > or =600 mg/kg/day. Metformin was also associated with evidence of minimal metabolic acidosis (increased serum lactate and beta-hydroxybutyric acid and decreased serum bicarbonate and urine pH) at doses > or =600 mg/kg/day. There were no significant sex differences in mean AUC(0-24) or C(max) nor were there significant differences in mean AUC(0-24) or C(max) following repeated dosing compared to a single dose. The no observable adverse effect level (NOAEL) was 200 mg/kg/day (mean AUC(0-24)=41.1 microg h/mL; mean C(max)=10.3 microg/mL based on gender average week 13 values). These effects should be taken into consideration when assessing potential toxicities of metformin in fixed dose combinations. PMID- 20004681 TI - H4 receptor antagonism exhibits anti-nociceptive effects in inflammatory and neuropathic pain models in rats. AB - The histamine H(4) receptor (H(4)R) is expressed primarily on cells involved in inflammation and immune responses. To determine the potential role of H(4)R in pain transmission, the effects of JNJ7777120, a potent and selective H(4) antagonist, were characterized in preclinical pain models. Administration of JNJ7777120 fully blocked neutrophil influx observed in a mouse zymosan-induced peritonitis model (ED(50)=17 mg/kg s.c., 95% CI=8.5-26) in a mast cell-dependent manner. JNJ7777120 potently reversed thermal hyperalgesia observed following intraplantar carrageenan injection of acute inflammatory pain (ED(50)=22 mg/kg i.p., 95% CI=10-35) in rats and significantly decreased the myeloperoxide activity in the carrageenan-injected paw. In contrast, no effects were produced by either H(1)R antagonist diphenhydramine, H(2)R antagonists ranitidine, or H(3)R antagonist ABT-239. JNJ7777120 also exhibited robust anti-nociceptive activity in persistent inflammatory (CFA) pain with an ED(50) of 29 mg/kg i.p. (95% CI=19-40) and effectively reversed monoiodoacetate (MIA)-induced osteoarthritic joint pain. This compound also produced dose-dependent anti allodynic effects in the spinal nerve ligation (ED(50)=60 mg/kg) and sciatic nerve constriction injury (ED(50)=88 mg/kg) models of chronic neuropathic pain, as well as in a skin-incision model of acute post-operative pain (ED(50)=68 mg/kg). In addition, the analgesic effects of JNJ7777120 were maintained following repeated administration and were evident at the doses that did not cause neurologic deficits in rotarod test. Our results demonstrate that selective blockade of H(4) receptors in vivo produces significant anti-nociception in animal models of inflammatory and neuropathic pain. PMID- 20004683 TI - Incidence and risk factors for the metabolic syndrome in an urban South European population. AB - OBJECTIVE: To estimate the incidence of the metabolic syndrome in a Portuguese sample and to evaluate its specific risk factors. METHODS: During a follow-up evaluation (May 2005-September 2008) of a Portuguese community cohort, 1377 participants were eligible for this study. Data on social, demographic, personal and family medical history and behavioral characteristics (including alcohol consumption and smoking habits) were collected. The metabolic syndrome was defined according to the AHA/NHLBI criteria Crude incidence rates were calculated for the total sample, by sex and age classes, and Poisson generalized linear models were used, with the default log link and offset in the variable time. RESULTS: The incidence rate of metabolic syndrome in this population was 47.2 /1000 person-years (95% CI: 41.2-53.9), similar in females and in males. The incidence of metabolic syndrome increased with age and in less educated participants. No significant association was observed between the occurrence of metabolic syndrome and the behavioral characteristics evaluated. High waist circumference presented the strongest association with the occurrence of the metabolic syndrome, even after adjustment for other confounders (IRR=1.68; 95% CI: 1.47-1.93). CONCLUSION: Older and less educated participants had a higher syndrome incidence. In this population, central obesity was the strongest predictor of metabolic syndrome. PMID- 20004682 TI - Inhibitory effect of ethanol extract of Magnolia officinalis and 4-O methylhonokiol on memory impairment and neuronal toxicity induced by beta amyloid. AB - The components of the herb Magnolia officinalis have exhibited antioxidant and neuroprotective activities. In this study, we investigated effects of ethanol extract of M.officinalis and its major component 4-O-methylhonokiol on memory dysfunction and neuronal cell damages caused by A beta. Oral pretreatment of ethanol extract of M. officinalis (2.5, 5 and 10mg/kg) and 4-O-methylhonokiol (1mg/kg) into drinking water for 5 weeks suppressed the intraventricular treatment of A beta(1-42) (0.5 microg/mouse, i.c.v.)-induced memory impairments. In addition, 4-O-methylhonokiol prevented the A beta(1-42)-induced apoptotic cell death as well as beta-secretase expression. 4-O-methylhonokiol also inhibited H(2)O(2) and A beta(1-42)-induced neurotoxicity in cultured neurons as well as PC12 cells by prevention of the reactive oxygen species generation. 4-O methylhonokiol also directly inhibited beta-secretase activity and A beta fibrilization in vitro. Thus, ethanol extract of M. officinalis may be useful for prevention of the development or progression of AD, and 4-O-methylhonokiol may be a major active component. PMID- 20004684 TI - Psychological distress is independently associated with physical inactivity in Israeli adults. AB - OBJECTIVES: Limited data exist on the link between psychological distress and regular physical activity in the general population. We examined the association of psychological distress with physical inactivity, independently of sociodemographic and behavioral factors in a large community sample. We also examined whether psychological distress is related to limitations in day-to-day activities due to emotional problems. METHODS: Information on physical activity, sociodemographic, and behavioral factors was obtained through telephone interviews of 5708 subjects aged > or = 21 years from a cross-sectional study-the first Israeli National Health Interview Survey (2003-2004). Psychological distress and limitations due to emotional problems were measured using the five item Mental Health scale (MHI-5) and role emotional scales derived from the SF-36 questionnaire. RESULTS: In multivariate analyses, high psychological distress level was associated with increased odds of physical inactivity among both men (adjusted OR=1.30, 95% CI=1.09-1.55) and women (adjusted OR=1.31, 95% CI=1.11 1.53). Psychological distress was strongly associated with limitations in day-to day activities due to emotional problems. CONCLUSIONS: In this cross-sectional study of adult men and women, psychological distress was independently associated with physical inactivity. Psychological distress may limit day-to-day activities in general and reduce the success of health promotion activities. These findings could help in identifying subjects facing difficulties in initiation and adherence to these activities. PMID- 20004685 TI - Monte Carlo simulation of mechanical unfolding of proteins based on a simple two state model. AB - Single molecule methods are becoming routine biophysical techniques for studying biological macromolecules. In mechanical unfolding of proteins, an externally applied force is used to induce the unfolding of individual protein molecules. Such experiments have revealed novel information that has significantly enhanced our understanding of the function and folding mechanisms of several types of proteins. To obtain information on the unfolding kinetics and the free energy landscape of the protein molecule from mechanical unfolding data, a Monte Carlo simulation based on a simple two-state kinetic model is often used. In this paper, we provide a detailed description of the procedure to perform such simulations and discuss the approximations and assumptions involved. We show that the appearance of the force versus extension curves from mechanical unfolding of proteins is affected by a variety of experimental parameters, such as the length of the protein polymer and the force constant of the cantilever. We also analyze the errors associated with different methods of data pooling and present a quantitative measure of how well the simulation results fit experimental data. These findings will be helpful in experimental design, artifact identification, and data analysis for single molecule studies of various proteins using the mechanical unfolding method. PMID- 20004686 TI - New insights in diagnosing Schistosoma myelopathy. AB - At present non-invasive tests for diagnosing Schistosoma myelopathy are sub optimal. We present a novel serological method, using paired liquor and serum samples, resulting in the diagnosis of Schistosoma myelopathy in a male patient with proximal muscle weakness. The patient recovered after praziquantel treatment. PMID- 20004687 TI - Sex patterns in diagnoses of tularaemia, Sweden 1997-2008. PMID- 20004688 TI - Advances in viral transneuronal tracing. AB - Powerful transneuronal tracing technologies exploit the ability of some neurotropic viruses to travel across neuronal pathways and to function as self amplifying markers. Two main classes of viral transneuronal tracers are available, derived from alpha-herpesviruses (Herpes Simplex virus type 1, Pseudorabies) and rabies virus. Depending on the virus type and strain, there are major differences with regard to host range, peripheral uptake, replication mechanisms, transport direction and specificity. While alpha-herpesviruses are the tracers of choice for studying autonomic innervation, rabies virus is the ideal tool for studying motor innervation, since its peripheral uptake occurs exclusively at motor endplates. Rabies virus is the only viral tracer that is entirely specific, as it moves exclusively across chemical synapses by strictly unidirectional (retrograde) transneuronal transfer without altering neuronal metabolism, allowing for the stepwise, time-dependent, identification of neuronal networks across an unlimited number of synapses. This review will highlight and contrast the different properties of these viral tracers, and summarize the methodological issues that are critical for the appropriate execution and interpretation of transneuronal tracing studies. Combinations of viral tracing with other methodologies will be evaluated. Emerging technologies, based on genetically modified herpes and rabies tracers, will be also discussed and put in perspective. PMID- 20004690 TI - A simple and rapid method of bacterial transformation. AB - Recently, a unique method for bacterial transformation using nanofibers to inoculate DNA has been developed by Naoto Yoshida and colleagues. We have verified the principle, transforming Escherichia coli, Yersinia enterocolitica and Acinetobacter baumannii, and have established a user-friendly protocol. A buffered suspension of sepiolite-an inexpensive, fibrous yet inoffensive mineral is mixed with bacteria and transforming DNA and the mixture directly spread on selective agar. PMID- 20004689 TI - IgG antibodies produced during subcutaneous allergen immunotherapy mediate inhibition of basophil activation via a mechanism involving both FcgammaRIIA and FcgammaRIIB. AB - The majority of human subjects who receive subcutaneous allergen immunotherapy (IT) develop decreased sensitivity to their allergens. Multiple factors may explain the efficacy of IT, some evidence support a role for allergen specific IgG antibodies. There is controversy whether such antibodies act by blocking allergen binding to IgE or initiation of active inhibitory signaling through low affinity IgG receptors (FcgammaRIIB) on mast cells and basophils. In this study, we addressed this question using peripheral blood from cat non-allergic, cat allergic, and immunotherapy-treated cat allergic subjects. Blood from subjects who received IT contain IgG antibodies that mediate inhibition of basophil activation by a mechanism that is blocked by antibodies specific for the inhibitory IgG receptor FcgammaRIIB. Surprisingly, inhibition was also blocked by aglycosylated, putatively non-FcR binding, antibodies that are specific for the FcgammaRIIA, suggesting a contribution of this receptor to the observed effect. Consistent with a cooperative effect, ex vivo basophils were found to express both IgG receptors. In other studies we found that basophils from subjects who were both chronically exposed to allergen and were producing both cat allergen specific IgE and IgG, are hyporesponsive to allergen. These studies confirm that IgG antibodies produced during IT act primarily by stimulation of inhibitory signaling, and suggest that FcgammaRIIA and FcgammaRIIB function cooperatively in activation of inhibitory signaling circuit. We suggest that under normal physiologic conditions in which only a small proportion of FcepsilonRI are occupied by IgE of a single allergen specificity, FcgammaRIIA co-aggregation may, by providing activated Lyn, be required to fuel activation of inhibitory FcgammaRIIB function. PMID- 20004691 TI - Decreased P300 current source density in drug-naive first episode schizophrenics revealed by high density recording. AB - Decreased P300 amplitude has been reported in schizophrenics during their first episode. The brain sources that contribute to this abnormality have not yet been well documented, and were investigated in the present study using high density EEG recordings. Nineteen drug-naive first episode schizophrenics were compared to 25 normal controls. Auditory P300 was elicited using an oddball paradigm. The brain sources of P300 ERP were reconstructed by performing low resolution of electromagnetic tomography (LORETA) analysis. No group difference in P300 latency was found. P300 amplitude was smaller for schizophrenics than for controls. Topographical analysis revealed that P300 amplitude reduction in schizophrenics was significant over left and medial regions of interest (ROIs). LORETA analysis of the P300 peak revealed that, the brain sources of P300 were symmetrically distributed over left and right hemispheres among the normal controls, but were asymmetrically distributed among the patients, with a reduction predominantly over the left temporal area. Statistical non-Parametric Mapping analysis identified 29 voxels of a significant group difference, which focused on left insula, left superior temporal gyrus (STG) and left postcentral gyrus (PCG). In addition, the mean P300 current source density over left insula, left STG and left PCG correlated inversely with the patients' Positive and Negative Syndrome Scale scores. The neural substrates that contributed to the decreased P300 amplitude in drug-naive first episode schizophrenia relatively focused on left STG and its nearby areas. These areas are probably involved in the pathogenesis of schizophrenia, and possible mechanisms for pathology need to be further clarified. PMID- 20004693 TI - Micelles based on HPMA copolymers. AB - Polymeric micelles have been under extensive investigation during the past years as drug delivery systems, particularly for anticancer drugs. They are formed by the self-assembly of amphiphilic block copolymers in aqueous solutions and have a spherical shape and a size in the nano-range (<200nm). Tumor accumulation of polymeric micelles upon intravenous administration can occur as a result of the leaky vasculature of tumor tissue (called the enhanced permeation and retention (EPR) effect).To benefit from the EPR effect, polymeric micelles need to have prolonged circulation times as well as high and stable drug loadings. Poly[N-(2 hydroxypropyl) methacrylamide] (pHPMA) is a hydrophilic polymer currently under investigation for its use in polymer-drug conjugates. Its biocompatibility, non immunogenicity and the possibility for functionalization are properties that resulted in broad pharmaceutical and biomedical applications, also in the micelle technology research. Being hydrophilic, it can serve as a micellar stealth corona, while it can also be modified with hydrophobic moieties to serve as a micellar core in which hydrophobic drugs can be solubilized and retained. HPMA based polymeric micelles have been showing very promising in vitro and in vivo results. This review summarizes the applications of pHPMA in the field of polymeric micelles, either serving as a micellar stealth corona, or, if hydrophobically rendered by derivatization, as a micellar core. PMID- 20004692 TI - Non-viral systemic delivery of Fas siRNA suppresses cyclophosphamide-induced diabetes in NOD mice. AB - A membrane receptor, Fas (CD95), and its ligand FasL have been considered as key players in diabetes pathogenesis. They are known to mediate interactions between beta cells and cytotoxic T cells, which results in apoptotic cell death. We hypothesized that the interruption of Fas-FasL interactions by suppressing Fas expression in beta cells would affect the development of diabetes. The effect of Fas-silencing siRNA (Fas siRNA) on diabetes development was evaluated in a cyclophosphamide (CY)-accelerated diabetes animal model after intravenous administration using a polymeric carrier, polyethylenimine (PEI). The systemic non-viral delivery of Fas siRNA showed significant delay in diabetes incidence up to 40 days, while the control mice treated with naked Fas siRNA, scrambled dsRNA, or PBS were afflicted with diabetes within 20 days. The retardation of diabetes incidence after the treatment of Fas siRNA may be due to the delayed progression of the pancreatic insulitis. In this study, the potential use of a non-viral carrier based siRNA gene therapy for the prevention of type-1 diabetes is demonstrated. PMID- 20004695 TI - Distribution of killer cell immunoglobulinlike receptors in the Macedonian population. AB - The aim of this study was to analyze killer immunoglobulinlike receptor (KIR) gene polymorphism in the Macedonian population. The study sample consists of 214 healthy unrelated individuals, aged 20-35 years. All individuals are of Macedonian origin and nationality, and residents of different geographic regions. The population genetics analysis package, Arlequin, was used for analysis of the data. We found that all 16 KIR genes were observed in the Macedonian population and framework genes KIR3DL3, KIR2DL4, and KIR3DL2 were present in all individuals. A total of 56 different KIR genotypes were found in the Macedonian population, based on the presence of 16 KIR genes. Neighbor-joining phylogenetic tree, constructed on the basis of standard genetic distances of KIR genes, shows that Macedonian population is in the same cluster with England West Midlands Indian Asian, Brazil SouthEast Caucasian, Romania Caucasians, Spain Basque, England West Midlands Caucasian, France Reunion, and Spain Granada populations. The frequency of KIR loci in Macedonian population shares several general features with other Caucasoid populations studied before. PMID- 20004694 TI - Estrogens induce visfatin expression in 3T3-L1 cells. AB - Visfatin is a 56 kDa protein that is overexpressed in pregnant women. Like insulin, 2 nM visfatin induced GLUT 4 translocation from the cytosolic fraction to the membrane in 3T3-L1 cells. We have previously reported that visfatin induces glucose uptake into 3T3-L1 cells. These two actions define visfatin as an insulinomimetic. Three estrogens are elevated in pregnancy. Estradiol, the predominant estrogen, estriol, produced by the fetal liver and the pro-estrogen progesterone are all higher during pregnancy than in nonparous women. 3T3-L1 cells were treated with 150 ng/ml estriol, 16 ng/ml estradiol or 190 ng/ml progesterone to reflect the circulating concentrations of these steroids during pregnancy. Estriol treatment produced a 2.5-fold increase in visfatin gene expression. Estradiol and progesterone had small but insignificant effects on visfatin gene expression. In a second experiment, cells were treated with a combination of all three steroids together at the same concentrations listed above. The combination treatment produced a 13-fold increase in visfatin gene expression. These data suggest that the estriol, estradiol and progesterone exert a synergistic effect on visfatin gene expression. Taken together these data suggest that visfatin may play a physiological role during pregnancy. Since visfatin potently and efficaciously induced GLUT 4 translocation in a cell culture model, any hypothetical role for visfatin in pregnancy should include the possibility that it may play a role in maternal/fetal glucose metabolism or distribution. Two possibilities present: either maternal visfatin is overexpressed as a protective response in the pregnant female to compensate for the insulin resistance that often accompanies pregnancy or the excess visfatin is a compensatory response to ensure adequate glucose delivery to the growing fetus. PMID- 20004696 TI - Atropine increases sevoflurane potency in cortical but not spinal networks during cholinergic overstimulation. AB - In the event of mass destruction with nerve agents a number of victims can be expected to suffer from symptoms of cholinergic overstimulation due to intoxication as well as from physical trauma. Since previous studies have demonstrated that cholinesterase inhibitors may reverse general anaesthesia in humans this scenario raises the question of how these patients can be anaesthetised in order to enable surgical interventions. A likely reason for this reversal is a reduction of anaesthetic potency by acetylcholine as observed for volatile anaesthetics in vitro. In order to test whether a combination of cholinergic antagonists with general anaesthetics improves their potency, we investigated the effects of clinically relevant concentrations of atropine on sevoflurane potency in cortical and spinal slice cultures during cholinergic overstimulation. As the spinal cord and neocortex are important substrates for general anaesthetics cultured spinal and cortical tissue slices were obtained from embryonic and newborn mice, respectively. Drug effects were assessed by extracellular voltage recordings of spontaneous action potential activity. Application of acetylcholine elevated spontaneous activity in neocortical and spinal slices. Atropine (10 nM) reduced discharge rates and reversed the increase of spontaneous activity induced by acetylcholine. In the presence of acetylcholine and atropine sevoflurane caused a concentration-dependent decrease of neuronal activity in neocortical (EC(50)=0.35+/-0.33 MAC) and spinal slices (EC(50)=0.43+/-0.03 MAC). Comparing our results with previous studies which investigated the effects of acetylcholine on anaesthetic potency it is concluded that small concentrations of atropine increase sevoflurane potency in cortical networks during cholinergic overstimulation. Thus, in a clinical setting, we recommend that anaesthetic drugs should be co-applied with atropine for adequate performance of general anaesthesia. PMID- 20004697 TI - Involvement of advanced glycation end-products, pentosidine and N(epsilon) (carboxymethyl)lysine, in doxorubicin-induced cardiomyopathy in rats. AB - In the pathogenesis of doxorubicin (DXR)-induced cardiomyopathy, oxidative stress appears to play an important role. It has been reported that pentosidine and N(epsilon)-(carboxymethyl)lysine (CML), advanced glycation end-products (AGEs), are formed by the combined processes of glycation and oxidation and play a significant role in the process of complications of diabetic mellitus. We investigated the potential involvement of AGE formation in DXR-induced cardiomyopathy in rats. Male Crl:CD(SD) rats received intravenous injection of DXR at 2mg/kg or saline once weekly for 8 weeks, with or without daily treatment with the AGE formation inhibitors, aminoguanidine (AG, 25 mg/kg/day, i.p.) and pyridoxamine (PM, 60 mg/kg/day, i.p.). Time-course experiments revealed significantly increased pentosidine and CML in the heart in the DXR group from Week 6. These findings coincided with a decrease in fractional shortening (FS), an index of cardiac function, and the development of cardiomyopathy characterized by vacuolated hypertrophic myocardial fibers. There was a significant correlation between the myocardial AGEs and FS or plasma cardiac troponin-I. Immunohistochemical staining showed localization of pentosidine to the cytoplasm of vacuolated myocardial cells. In DXR-treated rats, oxidative stress was enhanced prior to any observed increase in pentosidine and CML levels in the heart. Hyperglycemia was not observed throughout the study period. Intervention by AG or PM treatment ameliorated the functional and morphological changes induced by DXR in the heart, in addition to lowered myocardial pentosidine and CML levels. These results suggested that DXR accelerates the formation of pentosidine and CML in the heart through enhanced oxidative stress and that AGE formation is involved in DXR-induced cardiomyopathy. The findings may enable development of novel preventive therapies and predictive biomarkers of DXR induced cardiomyopathy. PMID- 20004698 TI - Systemic disorders and their influence on the development of dental hard tissues: a literature review. AB - OBJECTIVES: This report highlights the influence of a number of disorders with systemic physiological effects that impact on the development of dental hard tissues. It focuses in particular, on the pathological effects of systemic conditions with less well recognised, but no less important, impacts on dental development. Such conditions, include cystic fibrosis, HIV/AIDS, leukaemia, Alstrom syndrome, hypophosphatasia, Prader-Willi syndrome, Tricho-dento-osseous syndrome, tuberous sclerosis, familial steroid dehydrogenase deficiency and epidermolysis bullosa. These, along with developmental and environmental causes of enamel and dentine defects, are discussed and the possible aetiology of such effects are proposed. Furthermore, the dental management and long-term dental care of these patients is outlined. SOURCES: MEDLINE/PubMed. CONCLUSIONS: Enamel and dentine defects can present with a wide spectrum of clinical features and may be caused by a variety of factors occurring throughout tooth development from before birth to adulthood. These may include host traits, genetic factors, immunological responses to cariogenic bacteria, saliva composition, environmental and behavioural factors and systemic diseases. These diseases and their spectrum of clinical manifestations on the organs affected (including the dentition) require an increased knowledge by dental practitioners of the disease processes, aetiology, relevant treatment strategies and prognosis, and must encompass more than simply the management of the dental requirements of the patient. It is important that the impact of the disease and its treatment, particularly in respect of immunosuppression where dental interventions may become life threatening, is also taken into consideration. PMID- 20004699 TI - Attenuated Th1 induction by dendritic cells from mice deficient in the leukotriene B4 receptor 1. AB - Dendritic cells (DCs) are important antigen-presenting cells that control Th1- and Th2-type immunological reactions by releasing cytokines and interacting directly with T cells. Leukotriene B4 (LTB4), a classical proinflammatory lipid mediator for phagocytes, was recently identified as an important attractant for effector CD4(+) and CD8(+) T cells. However, little information is available on the roles of LTB4 and its receptor BLT1 in DCs. Here we show that functional BLT1 expressed in mouse bone marrow-derived DCs (BMDCs) plays important role in initiating Th1-type immune response. Detailed analyses using BMDCs revealed that BLT1-deficient DCs produced less IL-12p70 than WT DCs, leading to attenuated IFN gamma production in an allogeneic mixed lymphocyte reaction. Adoptive transfer of antigen-loaded BLT1-deficient DCs into naive WT mice induced a weakened Th1- and enhanced Th2-response in vivo compared to WT DCs. BLT1-deficient mice consistently showed much attenuated delayed-type hypersensitivity (DTH), in which Th1-type cellular responses play a key role, and popliteal lymph node cells of BLT1-deficient mice showed reduced production of Th1 cytokines after DTH induction compared to cells from WT mice. Thus, in addition to its role in inflammation, the LTB4-BLT1 axis is important in initiating Th1-type immunological reactions mediated by DCs. PMID- 20004700 TI - Effect of baicalein from Scutellaria baicalensis on prevention of noise-induced hearing loss. AB - Noise-induced hearing loss (NIHL) has been thought to primarily involve damage to the sensory hair cells of the cochlea via mechanical and metabolic mechanisms. This study examined the effects of baicalin, baicalein, and Scutellaria baicalensis (SB) extract against NIHL in a mouse model. Mice received oral treatment with SB, baicalin, baicalein beginning 30 min prior to noise exposure and continuing once daily throughout the study. Hearing threshold shift was assessed by auditory brain stem responses for 35 days following noise exposure. Central auditory function was evaluated by auditory middle latency responses. Cochlear function was determined based on transient evoked otoacoustic emissions. SB significantly reduced threshold shift, central auditory function damage, and cochlear function deficits, suggesting that SB may protect auditory function in NIHL and that the active constituent may be a flavonoid, baicalein. PMID- 20004701 TI - Melatonin is released in the third ventricle in humans. A study in movement disorders. AB - In order to determine sources and metabolism of melatonin in human cerebrospinal fluid (CSF), melatonin and 6-sulfatoxymelatonin (aMT6S) concentrations were measured in CSF sampled during neurosurgery in both lateral and third ventricles in patients displaying movement disorder (Parkinson's disease, essential tremor, dystonia or dyskinesia) and compared with their plasma levels. Previous determinations in nocturnal urine had showed that the patients displayed melatonin excretion in the normal range, compared with healthy controls matched according to age. A significant difference in melatonin concentration was observed between lateral and third ventricles, with the highest levels in the third ventricle (8.75+/-2.75 pg/ml vs. 3.20+/-0.33 pg/ml, p=0.01). CSF aMT6s levels were similar in both ventricles and of low magnitude, less than 5 pg/ml. They were not correlated with melatonin levels or influenced by the area of sampling. Melatonin levels were significantly higher in third ventricle than in the plasma, whereas there was no difference between plasma and lateral ventricle levels. These findings show that melatonin may enter directly the CSF through the pineal recess in humans. The physiological meaning of these data remains to be elucidated. PMID- 20004702 TI - Selective blockade of serotonin 2C receptor enhances Fos expression specifically in the striatum and the subthalamic nucleus within the basal ganglia. AB - Serotonin(2C) (5-HT(2C)) receptors are widely expressed in the basal ganglia, a group of brain regions involved in the control of motor behavior. However, it remains unclear whether their tonic influence on neuronal activity is distributed in these regions. We have addressed this question by measuring the product of the proto-oncogene c-Fos in rats after peripheral administration of the non-selective 5-HT antagonist mianserin, the 5-HT(2C/2B) antagonist SER-082 or the selective 5 HT(2C) antagonist SB 243213. The intraperitoneal administration of 1mg/kg of SB 243213 or SER-082, but not mianserin, enhanced Fos-immunoreactive cells in the subthalamic nucleus and the striatum, primarily its medial portion. None of these treatments significantly affected Fos expression in the external globus pallidus, the entopeduncular nucleus (the internal globus pallidus in primate) or the substantia nigra pars reticulata. The data suggest that selective blockade of 5 HT(2C) receptors is necessary to unmask a tonic regulation of neuronal activity by this receptor in the basal ganglia and that this effect is restricted to the two structures receiving cortical entries, the striatum and the subthalamic nucleus. PMID- 20004703 TI - Association between GBA L444P mutation and sporadic Parkinson's disease from Mainland China. AB - Mutations in GBA gene have been reported to be in patients with Parkinson's disease (PD) from different ethnic populations, including Taiwanese Chinese. To explore whether mutation in GBA is also associated with PD in Mainland China, we have now a case control study. The occurrence of the GBA L444P mutation was analyzed in an independent cohort of PD patients and controls from Mainland China. This mutation was present in 20/616 (3.2%) of PD compared with 1/411 (0.2%) of controls (odds ratio, OR=13.76, 95% Confidence interval, CI: 1.84 102.92, p=0.001). All carriers harbored the heterozygous genotype. In a subset analysis, the frequency of this mutation was higher both in early onset (EOPD) and late onset PD (LOPD) than in controls. However, no difference in clinical characteristics, such as gender, age at onset, onset symptoms, Hoehn-Yahr stage and UPDRS, was found between L444P carriers and non-carriers. In addition, we also explored the potential relationship between GBA L444P mutation and LRRK2 G2385R and R1628P variants in patients with PD. But no association was found, either. In conclusion, our data suggest that the GBA L444P mutation plays an important role in the development of PD also in Han-Chinese patients from Mainland China. PMID- 20004704 TI - The suprachiasmatic nucleus participates in food entrainment: a lesion study. AB - Daily feeding schedules entrain temporal patterns of behavior, metabolism, neuronal activity and clock gene expression in several brain areas and periphery while the suprachiasmatic nucleus (SCN), the biological clock, remains coupled to the light/dark cycle. Because bilateral lesions of the SCN do not abolish food entrained behavioral and hormonal rhythms it is suggested that food entrained and light entrained systems are independent of each other. Special circumstances indicate a possible interaction between the light and the food entrained systems and indicate modulation of SCN activity by restricted feeding. This study explores the influence of the SCN on food entrained rhythms. Food entrained temporal profiles of behavior, core temperature, corticosterone and glucose, as well as Fos and PER1 immunoreactivity in the hypothalamus and corticolimbic structures were explored in rats bearing bilateral SCN lesions (SCNX). In SCNX rats food anticipatory activity and the food entrained temperature and corticosterone increase were expressed with earlier onset and higher values than in intact controls. Glucose levels were lower in SCNX rats in all time points and SCNX rats anticipation to a meal induced higher c-Fos positive neurons in the hypothalamus, while a decreased c-Fos response was observed in corticolimbic structures. SCNX rats also exhibited an upregulation of the PER1 peak in hypothalamic structures, especially in the dorsomedial hypothalamic nucleus (DMH), while in some limbic structures PER1 rhythmicity was dampened. The present results indicate that the SCN participates actively during food entrainment modulating the response of hypothalamic and corticolimbic structures, resulting in an increased anticipatory response. PMID- 20004705 TI - Nitric oxide and interleukin-1beta mediate noradrenergic induced corticotrophin releasing hormone release in organotypic cultures of rat paraventricular nucleus. AB - Noradrenergic inputs from the brainstem are critical for the central stress response. It has been suggested that endogenous interleukin-1beta (IL-1beta) is involved in norepinephrine (NE)-induced release of corticotropin-releasing hormone (CRH) from the paraventricular nucleus of the hypothalamus (PVN). However, no IL-1 receptor on PVN CRH neurons has been identified. Therefore we hypothesized that the action of IL-1beta in the PVN requires downstream modulators that eventually lead to CRH release by PVN neurons. In the current study, we used organotypic cultures from neonatal rat PVN which display neuroendocrine characteristics suitable for in vitro studies. Pharmacological treatments with NE or IL-1beta elicited nitric oxide (NO) release from the PVN cultures, implying that local NO might be a candidate for modulating the action of IL-1beta. In addition, NE treatments significantly increased IL-1beta and CRH release. Treatment with IL-1beta or sodium nitroprusside also induced CRH release. Next, we also showed that either an IL-1 receptor antagonist or NOS inhibitor Nomega-nitro-L-arginine (L-NNA) attenuated the NE-induced CRH release. These results suggest that IL-1beta and NO are involved in NE-induced CRH release. Moreover, we found that application of L-NNA attenuated IL-1beta-induced CRH release, indicating that NO likely mediates this process. In summary, the current study demonstrates that IL-1beta plays a significant role in NE-induced CRH release, and that neuroendocrine response in the PVN may depend on local NO action. PMID- 20004706 TI - Nicotine induction of theta frequency oscillations in rodent hippocampus in vitro. AB - The hippocampus is an area important for learning and memory and exhibits prominent and behaviourally relevant theta (4-12 Hz) and gamma (30-100 Hz) frequency oscillations in vivo. Hippocampal slices produce similar types of oscillatory activity in response to bath-application of neurotransmitter receptor agonists. The medial septum diagonal band area (MS/DB) provides both a cholinergic and GABAergic projection to the hippocampus, and although it plays a major role in the generation and maintenance of the hippocampal theta rhythm in vivo, there is evidence for intrinsic theta generation mechanisms in the hippocampus, especially in area CA3. The aim of this study was to examine the role of the nicotinic receptor (nAChR) in the induction of oscillatory field activity in the in vitro preparation of the rat hippocampus. Bath-application of a low concentration of nicotine (1 muM) to transversely-cut hippocampal slices produced persistent theta-frequency oscillations in area CA3 of the hippocampus. These oscillations were reduced by both GABA(A) receptor antagonists and ionotropic glutamate receptor antagonists, indicating the involvement of local GABAergic and glutamatergic neurons in the production of the rhythmic theta activity. The nicotine-induced theta activity was inhibited by non-selective nAChR antagonists and partially by an alpha7* nAChR antagonist. The induction of theta frequency oscillations in CA3 by nicotine was mimicked alpha7* nAChR agonists but not by non-alpha7* nAChR agonists. In conclusion, theta activity in the hippocampus may be promoted by tonic stimulation of alpha7* nAChRs, possibly via selective stimulation of theta-preferring interneurons in the hippocampus that express post-synaptic alpha7* nAChRs. PMID- 20004708 TI - Brain volume regulation: osmolytes and aquaporin perspectives. AB - Cerebral water control is critical to maintain neuronal excitability, and to prevent injuries derived from brain swelling or shrinkage. The influence of aquaporins (AQPs) in the balance of water distribution between intracranial compartments is getting much experimental support. The importance of AQPs in fluid clearance during vasogenic brain edema seems well established but their role in cytotoxic swelling and in brain cell shrinkage is not known in detail. The main AQPs function as water channels anticipates their influence on cell volume changes as well as on the mechanisms of volume recovery, which include notably the osmolyte translocation across the cell membrane. Osmolyte fluxes permit the reestablishment of an osmotic balance and volume recovery in anisosmotic-elicited cell volume changes, but are also causal factors per se of brain cell swelling or shrinkage in pathological situations. This review aims to inform on the so far described functional interactions between AQPs and osmolyte fluxes and their volume-sensitive pathways. It also points to the coincidence of AQPs and activation of osmolyte fluxes in physiological and pathological conditions and to the importance of finding possible functional links between these two events, thus enlarging the possibilities via AQP manipulations, to prevent the adverse consequences of cell volume changes in brain. PMID- 20004707 TI - Replacement with GABAergic steroid precursors restores the acute ethanol withdrawal profile in adrenalectomy/gonadectomy mice. AB - The neurosteroid allopregnanolone (ALLO) is a progesterone metabolite that is one of a family of neuroactive steroids (NAS) that are potent positive allosteric modulators of gamma-aminobutyric acid(A) (GABA(A)) receptors. These GABAergic NAS are produced peripherally (in the adrenals and gonads) and centrally in the brain. Peripherally produced NAS modulate some effects of ethanol intoxication (e.g., anxiolytic, antidepressant, and anticonvulsant effects) in rodents. We have found that NAS also may be involved in the rebound neural hyperexcitability following a high ethanol dose. Removal of the adrenals and gonads (ADX/GDX) increased withdrawal severity following 4 g/kg ethanol, as measured by handling induced convulsions (HICs) in male and female DBA/2J mice. NAS are produced through the metabolism of progesterone (PROG), deoxycorticosterone (DOC), or testosterone, which can be blocked with the administration of finasteride (FIN), a 5alpha-reductase enzyme inhibitor. The current investigation was undertaken to clarify the step(s) in the biosynthetic NAS pathway that were sufficient to restore the acute ethanol withdrawal profile in ADX/GDX mice to that seen in intact animals. Male and female DBA/2J mice underwent ADX/GDX or SHAM surgery. After recovery, separate groups of animals were administered PROG, DOC, PROG+FIN, DOC+FIN, FIN, ALLO, ganaxalone (a synthetic ALLO derivative), corticosterone, or vehicle. Animals were then administered a 4 g/kg ethanol dose and allowed to undergo withdrawal. HICs were measured for 12 h and again at 24 h. The results indicate that replacement with PROG and DOC restored the withdrawal profile in ADX/GDX animals to SHAM levels, and that this effect was blocked with co administration of FIN. Administration of FIN alone increased the withdrawal profile in both SHAM and ADX/GDX males. These findings indicate that the increase in acute withdrawal severity after ADX/GDX may be due to the loss of GABAergic NAS, providing insight into the contribution of endogenous GABAergic NAS to ethanol withdrawal severity. PMID- 20004709 TI - A novel 65 kDa RNA-binding protein in squid presynaptic terminals. AB - A polyclonal antibody (C4), raised against the head domain of chicken myosin Va, reacted strongly towards a 65 kDa polypeptide (p65) on Western blots of extracts from squid optic lobes but did not recognize the heavy chain of squid myosin V. This peptide was not recognized by other myosin Va antibodies, nor by an antibody specific for squid myosin V. In an attempt to identify it, p65 was purified from optic lobes of Loligo plei by cationic exchange and reverse phase chromatography. Several peptide sequences were obtained by mass spectroscopy from p65 cut from sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) gels. BLAST analysis and partial matching with expressed sequence tags (ESTs) from a Loligo pealei data bank indicated that p65 contains consensus signatures for the heterogeneous nuclear ribonucleoprotein (hnRNP) A/B family of RNA-binding proteins. Centrifugation of post mitochondrial extracts from optic lobes on sucrose gradients after treatment with RNase gave biochemical evidence that p65 associates with cytoplasmic RNP complexes in an RNA-dependent manner. Immunohistochemistry and immunofluorescence studies using the C4 antibody showed partial co-labeling with an antibody against squid synaptotagmin in bands within the outer plexiform layer of the optic lobes and at the presynaptic zone of the stellate ganglion. Also, punctate labeling by the C4 antibody was observed within isolated optic lobe synaptosomes. The data indicate that p65 is a novel RNA binding protein located to the presynaptic terminal within squid neurons and may have a role in synaptic localization of RNA and its translation or processing. PMID- 20004710 TI - Anatomical tracer injections into the lower urinary tract may compromise cystometry and external urethral sphincter electromyography in female rats. AB - Physiological and anatomical investigations are commonly combined in experimental models. When studying the lower urinary tract (LUT), it is often of interest to perform both urodynamic studies and retrogradely labeled neurons innervating the peripheral target organs. However, it is not known whether the use of anatomical tracers for the labeling of, e.g. spinal cord neurons may interfere with the interpretation of the physiological studies on micturition reflexes. We performed cystometry and external urethral sphincter (EUS) electromyography (EMG) under urethane anesthesia in adult female rats at 5-7 days after injection of a 5% fluorogold (FG) solution or vehicle into the major pelvic ganglia (MPG) or the EUS. FG and vehicle injections into the MPG and EUS resulted in decreased voiding efficiency. MPG injections increased the duration of both bladder contractions and the inter-contractile intervals. EUS injections decreased EUS EMG bursting activity during voiding as well as increased both the duration of bladder contractions and the maximum intravesical pressure. In addition, the bladder weight and size were increased after either MPG or EUS injections in both the FG and vehicle groups. We conclude that the injection of anatomical tracers into the MPG and EUS may compromise the interpretation of subsequent urodynamic studies and suggest investigators to consider experimental designs, which allow for physiological assessments to precede the administration of anatomical tracers into the LUT. PMID- 20004711 TI - Molecular characterization of the intercalated cell masses of the amygdala: implications for the relationship with the striatum. AB - The intercalated cell masses of the amygdala consist of cell clusters located between the basolateral complex of the amygdala and its surrounding structures including the central nucleus of the amygdala and the external capsule. Although recent studies have revealed that the intercalated cell masses may play an important role in emotional learning and memory, there are only a few reports on its molecular characterization. We examined the expression patterns of transcription factors in the intercalated cell masses in late embryonic stage and postnatal rats, and non-human primates. Dlx5, Foxp2, Pbx3 and Meis2 were expressed in all subdivisions of the intercalated cell masses, while Ebf1, Nkx2.1 and Foxp1 were not. In contrast, Pax6 was only expressed in a small population of the main intercalated islands, but not in the medial or lateral cell clusters. In addition, few Pax6-positive neurons co-expressed Foxp2. Thus the intercalated cell masses do not contain a homogeneous population of neurons, in terms of their molecular constituents. Given that Foxp2, Pbx3 and Meis2 are preferentially expressed in distinct cell populations in the developing striatum, and that the intercalated cell masses of the amygdala appear to be a ventrocaudal expansion of the striatum, the intercalated neurons may share a common origin with some types of neurons located in the dorsal striatum. PMID- 20004712 TI - Vascular changes in epilepsy: functional consequences and association with network plasticity in pilocarpine-induced experimental epilepsy. AB - Angiogenesis and blood-brain-barrier (BBB) damage have been proposed to contribute to epileptogenesis and/or ictogenesis in experimental and human epilepsy. We tested a hypothesis that after brain injury angiogenesis occurs in the most damaged hippocampal areas with the highest need of tissue repair, and associates with formation of epileptogenic neuronal networks. We induced status epilepticus (SE) with pilocarpine in adult rats, and investigated endothelial cell proliferation (BrdU and rat endothelial cell antigen-1 (RECA-1) double labeling), vessel length (unbiased stereology), thrombocyte aggregation (thrombocyte immunostaining), neurodegeneration (Nissl staining), neurogenesis (doublecortin (DCX) immunohistochemistry), and mossy fiber sprouting (Timm staining) in the hippocampus at different time points post-SE. As functional measures we determined BBB leakage (quantified immunoglobulin G (IgG) immunostaining), and hippocampal blood volume (CBV) and flow (CBF) in vivo (magnetic resonance imaging, MRI). The total length of hippocampal blood vessels was decreased by 17% at 2 d after status epilepticus (SE) induced by pilocarpine in adult rats (P<0.05 as compared to controls) which was not accompanied by alterations in hippocampal blood volume (BV) and flow (BF). Number of proliferating endothelial cells peaked at 4 d post-SE and correlated with an increase in vessel length (r=0.900, P<0.05). Vessels length had recovered to control level or even higher at 2 wk post-SE, angiogenesis being most prominent in the CA3 (128% as compared to that in controls, P<0.05), and was associated with increased BV (178% as compared to that in controls, P<0.05). Enlargement of vessel diameter in the hippocampal fissure was associated with thrombocyte aggregation in distal capillaries. BBB was most leaky during the first 4 d post SE and increased IgG extravasation was observed for 60 d. Our data show that magnitude of endothelial cell proliferation is not associated with severity of acute post-SE neurodegeneration or formation of abnormal neuronal network. This encourages identification of molecular targets that initiate and maintain specific aspects of tissue reorganization, including preservation and proliferation of endothelial cells to reduce the risk of epileptogenesis and enhance recovery after brain injury. PMID- 20004713 TI - Perception and neural representation of size-variant human vowels in the Mongolian gerbil (Meriones unguiculatus). AB - Humans reliably recognize spoken vowels despite the variability of the sounds caused by the across-subject variability of the speakers' vocal tract. The vocal tract serves as a resonator which imprints a spectral envelope onto the sounds generated by the vocal folds. This spectral envelope contains not only information about the type of vocalization but also about the size of the speaker: the larger the speaker, the lower the formant frequencies of the spoken vowels. In a combined psychophysical and electrophysiological study in the Mongolian gerbil (Meriones unguiculatus), we investigated the perception and neural representation of human vowels spoken by speakers of different sizes. Gerbils trained to discriminate two standard vowels, correctly assigned vowels spoken from different-sized human speakers. Complementary electrophysiological recordings from neurons in the auditory brainstem, midbrain, and primary auditory cortex show that the auditory brainstem retains a truthful representation of the frequency content of the presented vowel sounds. A small percentage of neurons in the midbrain and auditory cortex, however, showed selectivity for a certain vowel type or vocal tract length which is not related to the pure-tone, frequency response area, indicative of a preprocessing stage for auditory segregation of size and structure information. PMID- 20004714 TI - Time-domain "wave" model of the human tympanic membrane. AB - Middle ear models have been successfully developed for many years. Most of those are implemented in the frequency domain, where physical equations are more easily derived. This is problematic, however, when it comes to model non-linear phenomena, especially in the cochlea, and because a frequency-domain implementation may be less intuitive. This research explores a different approach, based on a time-domain implementation, fitted to impedance data. It is adapted from a previous work for the cat and focuses here on the human ear: volume velocity samples are distributed uniformly in space and updated periodically to simulate the propagation of the sound wave in the ear. The modeling approach is simple, yet it can quantitatively reproduce the major characteristics of the human middle ear transmission, and can qualitatively capture forward and reverse power transmission - a key feature of this time domain implementation. These results suggest that complex, multi-modal propagation observed on the TM may not be critical to proper sound transmission along the ear. Besides, model predictions reveal that impedance and velocimetry measurements may be inconsistent with each other, hypothetically because velocimetry protocols could alter the middle ear. PMID- 20004715 TI - Interferon-gamma deficiency modifies the effects of a chronic stressor in mice: Implications for psychological pathology. AB - Pro-inflammatory cytokines promote behavioral and neurochemical variations similar to those evident following stressor exposure, and have been implicated in promoting depressive illness. Indeed, immunotherapeutic application of the cytokine, interferon-alpha, promoted depressive illness in cancer and hepatitis C patients. We assessed the possibility that another interferon cytokine family member, interferon-gamma (IFN-gamma), might contribute to the behavioral and biochemical alterations provoked by a chronic stressor regimen that has been used to model neuropsychiatric pathology in rodents. As predicted, IFN-gamma-deficient mice displayed basal differences in behavior (e.g., reduced open field exploration) and altered neurochemical activity (e.g., increased noradrenergic and serotonergic activity within the central amygdala), relative to their wild type counterparts. Moreover, stressor-induced elevations of corticosterone and the pro-inflammatory cytokine, tumor necrosis factor-alpha, were attenuated in IFN-gamma-deficient mice. Similarly, the IFN-gamma null mice were refractory to the chronic stressor-induced alterations of dopamine metabolism (within the prefrontal cortex, paraventricular nucleus of the hypothalamus and central amygdala) evident in wild-type mice. Yet, the chronic stressor provoked signs of anxiety (e.g., reduced open field exploration) and depression-like behavior (e.g., increased forced swim immobility, reduced consumption of a palatable solution) among both wild-type and IFN-gamma knockout mice alike, suggesting a dissociation of behavioral functioning from the stressor-induced alterations of immunological, hormonal and dopaminergic activity. Together, these data suggest a complex neurobehavioral phenotype, wherein IFN-gamma deletion engenders a state of heightened basal emotionality coupled with increased monoaminergic activity in the amygdala. At the same time, however, IFN-gamma deficiency appears to blunt some of the neurochemical, corticoid and cytokine alterations ordinarily associated with chronic stressor exposure. PMID- 20004717 TI - Colonization of a therapeutic spa with Legionella spp: a public health issue. AB - Traditional geothermally heated therapeutic spas, widely distributed in Europe, use water that is not disinfected. The colonization of therapeutic spas by Legionella spp. has been reported and several outbreaks and sporadic cases of legionellosis have been associated with the use of these facilities. We tested the effectiveness of hyperchlorination and the combination of hyperchlorination and ultraviolet light (UV) disinfection against legionellae which were persistently detected in the water used to supply the therapeutic spa. The hyperchlorination of the water distribution system was performed with a free residual chlorine concentration of 50 mg/l for 4h. An initial reduction in the numbers of legionellae was determined, but it rapidly increased to the original values. A UV unit was installed at the entrance to the spa's water distribution system and a new hyperchlorination procedure was performed immediately before UV irradiation (40 mJ/cm(2)), which was in use continuously for a nine-month period. While legionellae were not recovered during the irradiation period, these organisms were detected immediately after deactivation of the UV unit. We demonstrated that UV disinfection provides effective control of Legionella spp., with the advantage of being a method that, unlike chemical disinfectants, does not affect the physicochemical composition of the water. These findings suggest that UV irradiation, applied at key points in therapeutic spas, can be used to control colonization of water distribution systems by Legionella spp. PMID- 20004716 TI - RhoA/ROCK1 signaling regulates stress granule formation and apoptosis. AB - Cells form stress granules (SGs), in response to unfavorable environments, to avoid apoptosis, but it is unclear whether and how SG formation and cellular apoptosis are coordinately regulated. In this study we detected the small GTPase, Ras homolog gene family member A (RhoA), and its downstream kinase, Rho associated, coiled-coil containing protein kinase 1 (ROCK1), in SG, and found that their stress-induced activities were important for SG formation and subsequent global translational repression. Importantly, only activated RhoA and ROCK1 were sequestered into SG. Sequestration of activated ROCK1 into SG prevented ROCK1 from interacting with JNK-interacting protein 3 (JIP-3) and its activation of c-Jun N-terminal kinase (JNK), a pathway triggering apoptosis, thereby protecting cells from apoptosis. This study identifies a specific signaling pathway, mediated by RhoA and ROCK1, which determines cell fate by promoting SG formation or initiating apoptosis during stress. PMID- 20004718 TI - Wastewater bacterial communities bring together broad-host range plasmids, integrons and a wide diversity of uncharacterized gene cassettes. AB - To investigate the mobile gene pool present in wastewater environments, total community DNA was obtained from two distinct raw effluents: urban and slaughterhouse wastewaters. Bacterial community structure was evaluated by DGGE analysis of 16S rRNA gene fragments. Detection of broad-host-range plasmid sequences and integrase encoding genes was carried out through PCR and Southern hybridization. Gene cassette libraries were obtained using primers targeting consensus sequences that flank gene cassettes. Analysis of DGGE profiles revealed a complex and distinct bacterial community among effluents (similarity<25%). Despite that, All plasmid-specific sequences searched (rep for IncN, trfA for IncP-1 and oriV for IncQ and IncW) and integrase genes were present in both effluents. Gene cassettes recovered from clone libraries showed low homology with genes encoding putative enzymes involved in the metabolism of amino sugars, cell wall synthesis, motility, gene regulation, intercellular signalling and secretion pathways and in the synthesis of cellulose, folic acid and antibiotics. Additionally, in the majority of clones, no identifiable open reading frames homologues were found in the databases. According to our results, wastewater environments promote the development of bacterial communities that support and bring together different types of molecular elements that, in association, play a major role in bacterial adaptation and evolution. PMID- 20004719 TI - A history of spike trains as Point Processes in neural coding. AB - The history of viewing spike trains as point processes along time is that of the inference that spike trains participate in neural codings. A characteristic thought process, the "Conceptual Framework", present always and with sequential components, upholds the logic of this view. Early research in the 1920's introduced the view: experiments, though largely primitive, were designed and interpreted adequately and thus demonstrated convincingly this role. Modern research, starting around 1950, has contributed impressively. Essential was the mathematical Theory of Point Processes that permitted rigorous studies; computer implementation, though no substitute for clear thinking, was enormously helpful. Contributions, besides confirming the key inference, include wide-range train participation in neural codings, with many courses of action and formal implications involving neuronal performances individually and in networks, discharge forms, dynamic behaviors, critical Information and Communication issues, etc. PMID- 20004720 TI - Identification of a novel allele of peroxisome proliferator-activated receptor gamma (PPARG) and its association with resistance to Aeromonas salmonicida in Atlantic salmon (Salmo salar). AB - Bacterial and viral diseases are major problems in Atlantic salmon aquaculture, but may be challenged through selection of brood stock with enhanced survival to diseases. Today's selection strategy is based on controlled challenge tests using siblings of the breeding candidates, and is thus indirect. Direct trait records on breeding candidates can potentially be provided through identification of genetic variation linked to the susceptibility to the disease. Peroxisome proliferator-activated receptor gamma (PPARG) is a lipid-sensing transcription factor primarily known for inducing fat-accumulation in adipocytes, but also in lipid-accumulating macrophages, in mammalian species. Here we report a novel allele of PPARG, pparg-2, in Atlantic salmon. pparg-2 has an insertion of sixty nucleotides that encodes two additional copies of the almost perfect decapeptide motif, (F/C/Y)NHSPDR(S/N)HS, compared to the previously described pparg-1. pparg 1 contains six copies of this repeat unit whereas eight copies are present in the novel pparg-2 allele. pparg-2 mRNA was detectable in kidney and spleen of random Atlantic salmon samples. Here, we studied the effect of pparg-1 and pparg-2 on survival upon challenge to a highly virulent bacterium, Aeromonas salmonicida, causing furunculosis, and the virus causing infectious salmon anaemia (ISA), respectively, in a Norwegian aquaculture population of Atlantic salmon. ppar alleles were found to be significantly associated with survival upon challenge to A. salmonicida, but not to ISA. pparg-2 was the better allele in terms of survival in the challenge test for furunculosis, survival rates being 0.32, 0.40 and 0.42 for animals with the pparg-1,-1, pparg-1, -2 and pparg-2, -2 genotypes, respectively. We conclude that pparg-2 is in linkage disequilibrium (LD) with, or identical to, a locus contributing to different susceptibility to furunculosis in Atlantic salmon. PPARG was mapped to linkage group eight (LG8) but could only be positioned on the male linkage map since all the informative parents in the mapping families were males. This is the first report showing an association between pparg alleles and an enhanced immune response in fish. PMID- 20004721 TI - The association between metabolic rate, immune parameters, and growth performance of rainbow trout, Oncorhynchus mykiss (Walbaum), following the injection of a DNA vaccine alone and concurrently with a polyvalent, oil-adjuvanted vaccine. AB - This research demonstrates a significant increase in routine metabolic rate (RMR) following injection of a DNA vaccine concurrently with a polyvalent, oil adjuvanted vaccine. The increase in RMR was transient and associated with increased activity of both the non-specific and specific immune responses. Rainbow trout (Oncorhynchus mykiss) were injected with a DNA vaccine (DV), a commercially available polyvalent, oil-adjuvanted vaccine (AV), or the two vaccines in combination and sampled at 203, 305, and 406 days (dd) post-vaccine injection (pvi) for RMR and key immune parameters (serum lysozyme activity, serum neutralization antibody titres). The RMR of fish that received both the DV and the AV was significantly higher at 203 dd pvi, compared to fish from all other treatment groups which included the control, the AV, and the DV groups. The increased RMR corresponded to elevated levels of serum lysozyme activity and an earlier seroconversion of virus-specific neutralizing antibodies. To determine if growth performance was affected by the transient increase in RMR, specific growth rate (SGR), percent daily weight gain (WG), and feed conversion ratio (FCR) were determined at 798, 1204, and 1610 dd pvi. Although fish in all three vaccine groups showed significant increases in SGR and WG at 798 and 1610 dd pvi compared to the control group, the overall weight of the fish was not different at the end of the experiment. In summary, this study shows that concurrent injection of a DV and an AV transiently increases the RMR of rainbow trout and changes the manner in which the immune response occurs, but does not affect the overall growth performance of the fish. PMID- 20004722 TI - Immersion exposure of rainbow trout (Oncorhynchus mykiss) fry to wildtype Flavobacterium psychrophilum induces no mortality, but protects against later intraperitoneal challenge. AB - Flavobacterium psychrophilum, the causative agent of RTFS or rainbow trout fry syndrome, causes high mortality among hatchery reared rainbow trout (Oncorhynchus mykiss) fry in Europe and the USA. Despite several attempts, no efficient vaccines have yet been developed, the main obstacle being that the fry have to be vaccinated very early, i.e. around 0.2-0.5 g, where RTFS usually starts to give problems in the fish farms. Consequently, only oral or bath vaccines are relevant. Immersion of fry in inactivated or attenuated bacteria has resulted in RPS values of less than 50%. However, the results are biased by the fact that the fish have been challenged by intraperitoneal (ip) or subcutaneous (sc) injection against which an immersion/oral vaccine may not protect. Therefore, the present study was undertaken in order to investigate whether the presumably most potent immersion immunization, i.e. bathing in high titres of non-attenuated isolates of F. psychrophilum, was able to induce immunity to a subsequent ip challenge. Immersion in live bacteria for 30 or 50 min caused no mortality and protected a major fraction of the fry against challenges 26 and 47 days later with RPS values of 88.2 and 60.3%, respectively. Increased specific antibody titres suggested that adaptive immune mechanisms were involved in the protection. PMID- 20004723 TI - Neural processing of asynchronous audiovisual speech perception. AB - The temporal synchrony of auditory and visual signals is known to affect the perception of an external event, yet it is unclear what neural mechanisms underlie the influence of temporal synchrony on perception. Using parametrically varied levels of stimulus asynchrony in combination with BOLD fMRI, we identified two anatomically distinct subregions of multisensory superior temporal cortex (mSTC) that showed qualitatively distinct BOLD activation patterns. A synchrony defined subregion of mSTC (synchronous>asynchronous) responded only when auditory and visual stimuli were synchronous, whereas a bimodal subregion of mSTC (auditory>baseline and visual>baseline) showed significant activation to all presentations, but showed monotonically increasing activation with increasing levels of asynchrony. The presence of two distinct activation patterns suggests that the two subregions of mSTC may rely on different neural mechanisms to integrate audiovisual sensory signals. An additional whole-brain analysis revealed a network of regions responding more with synchronous than asynchronous speech, including right mSTC, and bilateral superior colliculus, fusiform gyrus, lateral occipital cortex, and extrastriate visual cortex. The spatial location of individual mSTC ROIs was much more variable in the left than right hemisphere, suggesting that individual differences may contribute to the right lateralization of mSTC in a group SPM. These findings suggest that bilateral mSTC is composed of distinct multisensory subregions that integrate audiovisual speech signals through qualitatively different mechanisms, and may be differentially sensitive to stimulus properties including, but not limited to, temporal synchrony. PMID- 20004724 TI - The amodal system for conscious word and picture identification in the absence of a semantic task. AB - Previous studies using explicit semantic tasks, such as category or similarity judgments, have revealed considerable neuroanatomical overlap between processing of the meaning of words and pictures. This result may have been influenced by the semantic executive control required by such tasks. We examined the degree of overlap while minimizing semantic executive demands. In a first fMRI experiment (n=28), we titrated word (35.3 ms, SD=9.6) and picture presentation duration (50.7 ms, SD=15.8) such that conscious stimulus identification became a stochastic process, with a 50% chance of success. Subjects had to indicate by key press whether or not they had been able to identify the stimulus. In a second fMRI experiment (n=19), the identification runs were followed by a surprise forced-choice recognition task and events were sorted on the basis of subsequent memory retrieval success rather than a subjective consciousness report. For both words and pictures, when stimulus processing exceeded the conscious identification threshold, the left occipitotemporal sulcus (OTS), intraparietal sulcus, inferior frontal junction, and middle third of the inferior frontal sulcus (IFS) were more active than when subjects had been unable to identify the stimulus. For both words and pictures, activity in two of these regions, IFS and OTS, predicted subsequent memory retrieval success. A Bayesian comparison revealed that the effective connectivity between IFS and the word- or picture specific systems was mainly mediated via its connections with OTS. The amodal nature of left OTS and IFS involvement in word and picture processing extends to tasks with minimal semantic executive demands. PMID- 20004726 TI - Effects of aging on default mode network activity in resting state fMRI: does the method of analysis matter? AB - Functional MRI (fMRI) of default mode network (DMN) brain activity during resting state is gaining attention as a potential non-invasive biomarker to diagnose incipient Alzheimer's disease. The aim of this study was to identify effects of normal aging on the DMN using different methods of fMRI processing and evaluation. METHODS: fMRI was acquired in 17 young and 21 old healthy subjects and the data were analyzed with (a) volumes of interest (VOI)-based signal time course and (b) independent component analyses (ICA). In the first approach, the strength of DMN region inter-connectivity (as expressed with correlation coefficients) was of primary interest, the second method provided a measure of the magnitude of DMN co-activation. RESULTS: The older subjects exhibited significantly lower DMN activity in the posterior cingulate (PCC, t-test P<.001) as well as a tendency to lower activity in all other DMN regions in comparison to the younger subjects. We found no significant effect of age on DMN inter connectivity. CONCLUSION: Effects of normal aging such as loss of PCC co-activity could be detected by ICA, but not by signal time course correlation analyses of DMN inter-connectivity. This either indicates lower sensitivity of inter connectivity measures to detect subtle DMN changes or indicate that ICA and time course analyses determine different properties of DMN co-activation. Our results, therefore, provide fundamental knowledge for a potential future use of functional MRI as biomarker for neurodegenerative dementias where diminished DMN activity needs to be reliably differentiated from that observed in health aging. PMID- 20004727 TI - Phylogeny and DNA barcoding of inquiline oak gallwasps (Hymenoptera: Cynipidae) of the Western Palaearctic. AB - We examine phylogenetic relationships within the Synergus complex of herbivorous inquiline gallwasps (Hymenoptera; Cynipidae; Synergini) associated with cynipid host galls on oak, a biologically diverse group whose genus-level morphological taxonomy has long been considered stable but whose species level taxonomy is problematic. We incorporate data for over 70% of recognised Western Palaearctic species in five morphology-based genera (Ceroptres, Saphonecrus, Synergus, Synophrus, Ufo), comprising sequence for two mitochondrial loci (coxI, cytb) and one nuclear locus (28S D2). In particular, we assess the evidence for monophyly of two long-established, morphology-defined sections within the genus Synergus that differ in a range of biological traits. To aid analyses of ecological interactions within oak cynipid communities, we also consider the utility of cytochrome oxidase I (coxI) DNA barcodes in the oak inquilines. In this assessment, we do not assume that species are delineated at a single threshold value of sequence divergence for a single gene, but examine concordance in the composition of molecular operational Taxonomic units (MOTUs) across a range of sequence divergences in each gene and across genes. We also assess the impact of sampling effort on MOTU stability. Phylogenetic reconstructions for all three loci support monophyly for Synergus and Synophrus, but reject monophyly for Saphonecrus and for the two sections within Synergus. The suites of traits associated with the two sections of the genus Synergus are thus homoplasious. All three loci also reject monophyly for three Synergus species (S. hayneanus, S. pallipes, S. umbraculus). Sequences for each locus identify robust MOTUs that are largely concordant across loci for a range of cut-off values. Though many MOTU's correspond to recognised Linnean species, there is significant, multigene disagreement between groupings supported by morphology and sequence data, with both allocation of different morphospecies to the same MOTU and allocation of the same morphospecies to multiple MOTUs, regardless of cut-off value. Our results imply that while DNA barcoding has considerable utility within this group, morphology-based identification needs major revision at both genus and species levels. Further, lifehistory traits currently attributed to single morphospecies probably confound attributes of multiple lineages. Revealing patterns of character state evolution in Synergus requires collection of new host association and life history data explicitly linked to DNA barcode data for the specimens concerned. PMID- 20004725 TI - Family history of alcoholism mediates the frontal response to alcoholic drink odors and alcohol in at-risk drinkers. AB - Although a family history of alcoholism is the strongest risk factor for developing alcohol dependence, there are few studies of the association between familial alcoholism and the human brain's reward system activity. We used functional magnetic resonance imaging (fMRI) to determine how family history affects the brain's response to subjects' preferred alcoholic drink odors (AO) as compared to appetitive control odors (ApCO). Fourteen non-dependent heavy drinkers (HD) who were family history positive (FHP) participated, as did 12 HD who were family history negative (FHN). Subjects were imaged under both alcohol intoxication and placebo, using intravenous infusion and pharmacokinetic modeling to target a blood alcohol level of 50 mg%. Under placebo, HD-FHP had a larger medial frontal [AO>ApCO] effect than did HD-FHN. Alcohol intoxication dampened this response in the HD-FHP but potentiated it in the HD-FHN. This suggests that a family history of alcoholism and brain exposure to alcohol interact in heavy drinkers to differentially affect how the brain responds to alcohol cues. PMID- 20004728 TI - Molecular phylogeny of euglyphid testate amoebae (Cercozoa: Euglyphida) suggests transitions between marine supralittoral and freshwater/terrestrial environments are infrequent. AB - Marine and freshwater ecosystems are fundamentally different regarding many biotic and abiotic factors. The physiological adaptations required for an organism to pass the salinity barrier are considerable. Many eukaryotic lineages are restricted to either freshwater or marine environments. Molecular phylogenetic analyses generally demonstrate that freshwater species and marine species segregate into different sub-clades, indicating that transitions between these two environments occur only rarely in the course of evolution. It is, however, unclear if the transitions between freshwater and environments characterized by highly variable salinities, such as the marine supralittoral zone, are also infrequent. Here, we use testate amoebae within the Euglyphida to assess the phylogenetic interrelationships between marine supralittoral and freshwater taxa. Euglyphid testate amoebae are mainly present in freshwater habitats but also occur in marine supralittoral environments. Accordingly, we generated and analyzed partial SSU rRNA gene sequences from 49 new marine/supralittoral and freshwater Cyphoderiidae sequences, 20 sequences of the Paulinellidae, Trinematidae, Assulinidae, and Euglyphidae families as well as 21 GenBank sequences of unidentified taxa derived from environmental PCR surveys. Both the molecular and morphological data suggest that the diversity of Cyphoderiidae is strongly underestimated. The results of our phylogenetic analyses demonstrated that marine supralittoral and freshwater euglyphid testate amoeba species are segregated into distinct sub-clades, suggesting that transitions between these two habitats occurred only infrequently. PMID- 20004729 TI - Red drifters and dark residents: the phylogeny and ecology of a Plio-Pleistocene dragonfly radiation reflects Africa's changing environment (Odonata, Libellulidae, Trithemis). AB - In the last few million years, tropical Africa has experienced pronounced climatic shifts with progressive aridification. Such changes must have had a great impact on freshwater biota, such as Odonata. With about forty species, Trithemis dominates dragonfly communities across Africa, from rain-pools to streams, deserts to rainforests, and lowlands to highlands. Red-bodied species tend to favor exposed, standing and often temporary waters, have strong dispersal capacities, and some of the largest geographic ranges in the genus. Those in cooler habitats, like forest streams, are generally dark-bodied and more sedentary. We combined molecular analyses of ND1, 16S, and ITS (ITSI, 5.8S, and ITSII) with morphological, ecological, and geographical data for 81% of known Trithemis species, including three Asian and two Madagascan endemics. Using molecular clock analyses, the genus's origin was estimated 6-9Mya, with multiple lineages arising suddenly around 4Mya. Open stagnant habitats were inferred to be ancestral and the rise of Trithemis may have coincided with savannah-expansion in the late Miocene. The adaptation of red species to more ephemeral conditions leads to large ranges and limited radiation within those lineages. By contrast, three clades of dark species radiated in the Plio-Pleistocene, each within distinct ecological confines: (1) lowland streams, (2) highland streams, and (3) swampy habitats on alternating sides of the Congo-Zambezi watershed divide; together giving rise to the majority of species diversity in the genus. During Trithemis evolution, multiple shifts from open to more forested habitats and from standing to running waters occurred. Allopatry by habitat fragmentation may be the dominant force in speciation, but possibly genetic divergence across habitat gradients was also involved. The study demonstrates the importance of combining ecological and phylogenetic data to understand the origin of biological diversity under great environmental change. PMID- 20004730 TI - Combination of molecular data support the existence of three main lineages in the phylogeny of aphids (Hemiptera: Aphididae) and the basal position of the subfamily Lachninae. AB - The first molecular studies on the phylogeny of aphids (Hemiptera: Aphididae) bumped into a striking lack of phylogenetic structure for taxa levels higher than tribe, probably as a consequence of the rapid adaptive radiation that this group of insects went through during the Late Cretaceous. Here we present a new attempt to infer the relationships between major aphid taxa by the separate and combined analysis of two nuclear sequences (the long-wavelength opsin gene and the elongation factor 1 alpha gene) and two mitochondrial sequences (the genes encoding the subunit 6 of the F-ATPase and the subunit II of the cytochrome oxidase). Our results confirm previous results with the grouping of the subfamilies analysed in three main lineages, that are named A+D (subfamilies Aphidinae, Calaphidinae, Chaitophorinae, Drepanosiphinae and Pterocommatinae), E+T (subfamilies Anoeciinae, Eriosomatinae, Hormaphidinae, Mindarinae and Thelaxinae) and L (subfamily Lachninae). Furthermore, phylogenetic reconstructions generally support the early branching of the subfamily Lachninae in the phylogeny of aphids. Although some relationships among subfamilies inside lineages are not highly supported, our results are compatible with a scenario for the evolution of aphid life cycles with only four transitions of feeding from gymnosperms to angiosperms and two origins of host alternation. PMID- 20004731 TI - Mitochondrial genome of Thais clavigera (Mollusca: Gastropoda): affirmation of the conserved, ancestral gene pattern within the mollusks. AB - Class Gastropoda includes a large number of described species, many with extensively rearranged mitochondrial genomes. We sequenced the mitogenome of the rock shell, Thais clavigera (Gastropoda: Muricidae), an intertidal snail, using long PCR with primers designed on the basis of expressed sequence tags. The mitogenome of T. clavigera consists of 2 rRNAs, 22 tRNAs, and 13 protein-coding genes, but no control region. Structural comparisons revealed that the order Sorbeoconcha, including T. clavigera, have nearly identical mitochondrial gene patterns. However, they have an inversion between a tRNA(Phe)-tRNA(Glu) cluster that comprises 21 genes, but most of the remaining structure is similar to the putative mollusk ground pattern. These findings will provide a better insight into mitochondrial gene rearrangement over the course of gastropod evolution. PMID- 20004732 TI - A vacuum-vortex technique for preparation of anoxic solutions or liquid culture media in small volumes for cultivating methanogens or other strict anaerobes. AB - A highly efficient method is described for producing at room temperature anoxic solutions of 50 ml or less in test tubes or serum vials by combining negative pressure with strong vortexing so that the liquid-surface, gas exchange area is increased by orders of magnitude. Liquid media suitable for the cultivation of methanogens may be rendered anoxic after three short vacuum-vortex steps. PMID- 20004733 TI - Metabolic engineering of omega-3 long-chain polyunsaturated fatty acids in plants using an acyl-CoA Delta6-desaturase with omega3-preference from the marine microalga Micromonas pusilla. AB - Long-chain (> or = C20) polyunsaturated fatty acids (LC-PUFA) EPA and DHA (20:5(Delta5,8,11,14,17) and 22:6(Delta4,7,10,13,16,19)) have well-documented health benefits against coronary heart disease, rheumatoid arthritis and other disorders. Currently, the predominant sources of these fatty acids are marine fish and algal oils, but research is being conducted to ensure that a sustainable, land-based production system can be developed. We here describe the metabolic engineering of an artificial pathway that produces 26% EPA in leaf triacylglycerol using a newly-identified Delta6-desaturase from the marine microalga Micromonas pusilla. We also demonstrate that this enzyme appears to function as an acyl-CoA desaturase that has preference for omega3 substrates both in planta and in yeast. Phylogenetic analysis indicates that this desaturase shares highly conserved motifs with previously described acyl-CoA Delta6 desaturases. PMID- 20004734 TI - A synthetic analog of 15-epi-lipoxin A4 inhibits human monocyte apoptosis: involvement of ERK-2 and PI3-kinase. AB - Human monocytes play a central function in several steps of the immune response and the process involved in regulating their survival are critical to population control. Lipoxins are lipid mediators and members of the eicosanoid family that exhibit selective stimulatory but nonphlogistic activities in mononuclear cells. In this study, we investigated the effects of 15-epi-16-(para-fluoro)phenoxy LXA(4) (ATL-1), a synthetic analog of 15-epi-lipoxin A(4), in human monocytes survival and apoptosis. ATL-1 concentration-dependently increased monocyte survival, as a consequence of cell apoptosis reduction by the analog. Treatment of these cells with PD98059 or LY294002 blocked ATL-1 effects, indicating the involvement of ERK-2 and PI3-K, both pathways associated with cell survival. Confirming the activation of these pathways, we demonstrated an increase in ERK-2 nuclear translocation and Akt phosphorylation. Furthermore, we showed that ATL-1 inhibits Bax translocation to the mitochondria. These results confirm a cytoprotective effect of lipoxins in monocytes and might contribute to the elucidation of the mechanisms associated with the resolution phase of the inflammatory process in different pathophysiological events. PMID- 20004736 TI - Twenty Sox, twenty years. PMID- 20004735 TI - Immune responses to Epstein-Barr virus: molecular interactions in the virus evasion of CD8+ T cell immunity. AB - Persistent viruses have mechanisms for modulating the host immune responses that are essential for achieving a lifelong virus-host balance while minimizing the viral pathogenicity. Here we review some of the immune-modulating mechanisms evolved by the ubiquitous but potentially oncogenic Epstein-Barr virus, with particular emphasis on the molecular mechanisms of genes interfering with HLA class I antigen presentation. PMID- 20004737 TI - Adult human brain cell culture for neuroscience research. AB - Studies of the brain have progressed enormously through the use of in vivo and in vitro non-human models. However, it is unlikely such studies alone will unravel the complexities of the human brain and so far no neuroprotective treatment developed in animals has worked in humans. In this review we discuss the use of adult human brain cell culture methods in brain research to unravel the biology of the normal and diseased human brain. The advantages of using adult human brain cells as tools to study human brain function from both historical and future perspectives are discussed. In particular, studies using dissociated cultures of adult human microglia, astrocytes, oligodendrocytes and neurons are described and the applications of these types of study are evaluated. Alternative sources of human brain cells such as adult neural stem cells, induced pluripotent stem cells and slice cultures of adult human brain tissue are also reviewed. These adult human brain cell culture methods could benefit basic research and more importantly, facilitate the translation of basic neuroscience research to the clinic for the treatment of brain disorders. PMID- 20004738 TI - Simvastatin activates Akt/glycogen synthase kinase-3beta signal and inhibits caspase-3 activation after experimental subarachnoid hemorrhage. AB - This study was designed to explore the role of simvastatin and its effects on the Akt/GSK3beta survival signal and apoptosis pathway after experimental subarachnoid hemorrhage (SAH). SAH was induced by blood injection into the cisterna magna in New Zealand white rabbits. Increased expression of phospho-Akt and phospho-GSK3beta was observed in brain tissue after SAH. Apoptosis and related proteins, including P53, apoptosis-inducing factor (AIF), cytochrome C, and cleaved caspase-3, were also activated. Simvastatin, at both low dose (10 mg/kg) and high dose (40 mg/kg), further increased expression of phospho-Akt and phospho-GSK3beta, decreased activation of caspase-3, and inhibited apoptosis. Preserved blood-brain barrier and attenuated brain edema were observed following simvastatin treatment. In addition, the neuroprotective effects of simvastatin were blocked by wortmannin (2.5 microg/kg/min), an irreversible PIK3 inhibitor. P53, AIF, and cytochrome C were not affected by simvastatin treatment. Findings from the present study suggest that simvastatin ameliorates acute brain injury after SAH. The potential mechanisms of action include activation of the Akt/GSK3beta survival signal and inhibition of caspase-dependent apoptosis pathway. PMID- 20004740 TI - Effect of adapted physical activity on health-related quality of life among hospitalized children and adolescents (the ACTIV'HOP randomized controlled trial): design and methods. AB - INTRODUCTION: In promoting the health of chronically ill children, the hospital should help children cope both medically and psychosocially with their disease. The University Children's Hospital of Nancy proposes adapted physical activity for hospitalized children, but the practice during hospitalization is not well developed or understood. The ACTIV'HOP study aims to assess the effects of adapted physical activity sessions for hospitalized children in terms of 1) health-related quality of life (HRQoL) and 2) satisfaction with hospital care. RESEARCH DESIGN: ACTIV'HOP is an individual randomized controlled trial of patients admitted to the University Children's Hospital of Nancy. The inclusion criteria are 1) age 5 to 18 years, 2) hospitalized for at least 3 days, and 3) no medical contra-indication to practice adapted physical activity. HYPOTHESIS: Children and adolescents practicing adapted physical activity during hospitalization have a better HRQoL on hospital discharge and better satisfaction with care. OUTCOMES: HRQoL assessed by the Child Health Questionnaire on the first and last day of hospitalization, and satisfaction with care assessed by the Patients Judgment of Hospital Quality questionnaire were sent to the family home 11 to 15 days after hospital discharge. The French validated version of these questionnaires is used. PMID- 20004741 TI - Rank-Minimization for balanced assignment of subjects in clinical trials. AB - Minimization (M) is the most popular algorithm for balancing large numbers of subject variables in treatment groups of small clinical trials. However, its use has been limited because of its complexity, vulnerability to selection bias and lack of a generally accepted method for statistical analysis of the data. Rank Minimization (RM) is a promising new algorithm. It is less complex since it does not require unique programming for each clinical trial to convert continuous to categorical variables. In this study RM is compared to M for balance of variables and vulnerability to selection bias in 1000 simulated trials using 200 subjects with 15 continuous variables. With RM there were no instances of significant imbalance to cause rejection of the null hypothesis, i.e. a Student's t> or =2, although it occurred in 0.4% of the 15000 tests for M. For moderate imbalance, i.e. 1< or = t < 2, the figures were 3% (RM) and 12% (M). The probability of guessing the next assignment was 0.636 (RM) and 0.683 (M). The smaller figure is superior to that of restricted randomization in blocks of five per treatment group. Improvement in balance, a decrease in vulnerability to selection bias and ease of application along with improvements in the statistical analysis should result in the general acceptance of RM for assigning subjects to treatment groups in clinical trials. PMID- 20004739 TI - A multi-center, randomized, controlled trial of parenteral nutrition titrated to resting energy expenditure in children undergoing hematopoietic stem cell transplantation ("PNTREE"): rationale and design. AB - BACKGROUND: Children undergoing hematopoietic stem cell transplantation (HSCT) frequently require prolonged courses of parenteral nutrition (PN) as a consequence of gastrointestinal dysfunction related to preparative chemotherapy and radiation. PN has been associated with shorter engraftment time and decreased mortality during HSCT, however, it is also linked with complications, including infections, liver disease, and metabolic disturbances. Some of these complications may be a result of providing PN in excess of nutrient requirements. We previously described significant reductions in resting energy expenditure (REE), as measured by indirect calorimetry, over the course of HSCT. We also documented a decline in mid-arm muscle area, suggesting depletion of muscle mass, while triceps skinfold, a marker of fat stores, was unchanged. These results suggested the need for further study of energy expenditure, body composition and nutritional intake in this group of high risk patients. DESIGN AND HYPOTHESIS: We hypothesize that changes in body composition affect REE during HSCT, and that standard nutritional support may lead to overfeeding. We are performing a randomized controlled trial of parenteral nutrition among children undergoing allogeneic HSCT. Subjects are randomized to receive PN designed to provide 100% of measured REE, or standard PN, i.e., 140% of estimated energy expenditure. The primary outcome variable is change in percent body fat. Secondary outcomes include glycemic control and frequency of infections, changes in REE and body composition. CONCLUSION: This study will provide unique and comprehensive nutritional data and its results will guide nutritional therapy for children undergoing HSCT and possibly other catabolic patients. PMID- 20004743 TI - Identity of the immunomodulatory proteins from garlic (Allium sativum) with the major garlic lectins or agglutinins. AB - Garlic (Allium sativum), an important medicinal spice, displays a plethora of biological effects including immunomodulation. Although some immunomodulatory proteins from garlic have been described, their identities are still unknown. The present study was envisaged to isolate immunomodulatory proteins from raw garlic, and examine their effects on certain cells of the immune system (lymphocytes, mast cells, and basophils) in relation to mitogenicity and hypersensitivity. Three protein components of approximately 13 kD (QR-1, QR-2, and QR-3 in the ratio 7:28:1) were separated by Q-Sepharose chromatography of 30 kD ultrafiltrate of raw garlic extract. All the 3 proteins exhibited mitogenic activity towards human peripheral blood lymphocytes, murine splenocytes and thymocytes. The mitogenicity of QR-2 was the highest among the three immunomodulatory proteins. QR-1 and QR-2 displayed hemagglutination and mannose-binding activities; QR-3 showed only mannose-binding activity. Immunoreactivity of rabbit anti-QR-1 and anti-QR-2 polyclonal antisera showed specificity for their respective antigens as well as mutual cross-reactivity; QR-3 was better recognized by anti-QR-2 (82%) than by anti-QR-1 (55%). QR-2 induced a 2-fold higher histamine release in vitro from leukocytes of atopic subjects compared to that of non-atopic subjects. In all functional studies, QR-2 was more potent compared to QR-1. Taken together, all these results indicate that the two major proteins QR-2 and QR-1 present in a ratio of 4:1 in raw garlic contribute to garlic's immunomodulatory activity, and their characteristics are markedly similar to the abundant Allium sativum agglutinins (ASA) I and II, respectively. PMID- 20004742 TI - Auto/paracrine control of inflammatory cytokines by acetylcholine in macrophage like U937 cells through nicotinic receptors. AB - Although acetylcholine (ACh) is well known for its neurotransmitter function, recent studies have indicated that it also functions as an immune cytokine that prevents macrophage activation through a 'cholinergic (nicotinic) anti inflammatory pathway'. In this study, we used the macrophage-like U937 cells to elucidate the mechanisms of the physiologic control of cytokine production by auto/paracrine ACh through the nicotinic class of ACh receptors (nAChRs) expressed in these cells. Stimulation of cells with lipopolysaccharide up regulated expression of alpha1, alpha4, alpha5, alpha7, alpha10, beta1 and beta3 subunits, down-regulated alpha6 and beta2 subunits, and did not alter the relative quantity of alpha9 and beta4 mRNAs. Distinct nAChR subtypes showed differential regulation of the production of pro- and anti-inflammatory cytokines. While inhibition of the expression of the TNF-alpha gene was mediated predominantly by the alpha-bungarotoxin sensitive nAChRs, that of the IL-6 and IL 18 genes-by the mecamylamine-sensitive nAChRs. Both the Mec- and alphaBtx sensitive nAChRs regulated expression of the IL-1beta gene equally efficiently. Upregulation of IL-10 production by auto/paracrine ACh was mediated predominantly through alpha7 nAChR. These findings offer a new insight on how nicotinic agonists control inflammation, thus laying a groundwork for the development of novel immunomodulatory therapies based on the nAChR subtype selectivity of nicotinic agonists. PMID- 20004744 TI - Kawasaki disease: aetiopathogenesis and therapeutic utility of intravenous immunoglobulin. AB - Kawasaki disease (KD) is an acute febrile childhood vasculitis, associated with the development of coronary artery abnormalities in 25-30% of untreated patients. The aetiopathogenesis is not well known but it is accepted that an undefined infectious trigger in genetically predisposed individuals results in the disease. KD is characterized by an endothelial cell injury, which could be due to abnormal cytokine production and to generation of cytotoxic antibodies against the endothelial cells. Intravenous immunoglobulin IVIG is an effective treatment in preventing the occurrence of coronary artery abnormalities in KD. Several mechanisms may explain the anti-inflammatory effects of IVIG in this disease. They include modification of the cytokine balance, and alteration on both the differentiation and the function of monocytes/macrophages, neutrophils and lymphocytes. PMID- 20004745 TI - Injectable, rapid gelling and highly flexible hydrogel composites as growth factor and cell carriers. AB - A family of injectable, rapid gelling and highly flexible hydrogel composites capable of releasing insulin-like growth factor (IGF-1) and delivering mesenchymal stromal cell (MSC) were developed. Hydrogel composites were fabricated from Type I collagen, chondroitin sulfate (CS) and a thermosensitive and degradable hydrogel copolymer based on N-isopropylacrylamide, acrylic acid, N acryloxysuccinimide and a macromer poly(trimethylene carbonate)-hydroxyethyl methacrylate. The hydrogel copolymer was gellable at body temperature before degradation and soluble at body temperature after degradation. Hydrogel composites exhibited LCSTs around room temperature. They could easily be injected through a 26-gauge needle at 4 degrees C, and were capable of gelling within 6s at 37 degrees C to form highly flexible gels with moduli matching those of the rat and human myocardium. The hydrogel composites showed good oxygen permeability; the oxygen pressure within the hydrogel composites was similar to that in the air. The effects of collagen and CS contents on LCST, gelation time, injectability, mechanical properties and degradation properties were investigated. IGF-1 was loaded into the hydrogel composites for enhanced cell survival/growth. The released IGF-1 remained bioactive during a 2-week release period. Small fraction of CS in the hydrogel composites significantly decreased IGF-1 release rate. The release kinetics appeared to be controlled mainly by hydrogel composite water content, degradation and interaction with IGF-1. Human MSC adhesion on the hydrogel composites was comparable to that on the tissue culture plate. MSCs were encapsulated in the hydrogel composites and were found to grow inside during a 7-day culture period. IGF-1 loading significantly accelerated MSC growth. RT-PCR analysis demonstrated that MSCs maintained their multipotent differentiation potential in hydrogel composites with and without IGF 1. These injectable and rapid gelling hydrogel composites demonstrated attractive properties for serving as growth factor and cell carriers for cardiovascular tissue engineering applications. PMID- 20004746 TI - In vitro degradation and mechanical integrity of Mg-Zn-Ca alloy coated with Ca deficient hydroxyapatite by the pulse electrodeposition process. AB - The key to manufacturing magnesium-based alloys that are suitable as biodegradable orthopaedic implants is how to adjust their degradation rates and mechanical integrity in the physiological environment. In this study, to solve this challenge, a soluble Ca-deficient hydroxyapatite (Ca-def HA) coating was deposited on an Mg-Zn-Ca alloy substrate by pulse eletrodeposition. This deposition can be demonstrated by X-ray diffractometry and energy dispersion spectroscopy analyses, and the Ca/P atomic ratio of as-deposited coating is about 1.33 (within the range from 1.33 to 1.65). By regulating the appropriate pulse amplitude and width, the Ca-def HA coating shows better adhesion to Mg-Zn-Ca alloy, whose lap shear strength is increased to 41.8+/-2.7 MPa. Potentiodynamic polarization results in Kokubo's simulated body fluid (SBF) indicate that the corrosion potential of Mg alloy increases from -1645 to -1414 mV, while the corrosion current density decreases from 110 to 25 microA/cm(2), which illustrates that the Ca-def HA coating improves the substrate corrosion resistance significantly. Since orthopaedic implants generally serve under conditions of stress corrosion, the mechanical integrity of the Mg-Zn-Ca alloy was measured using the slow strain rate tensile (SSRT) testing technique in SBF. The SSRT results show that the ultimate tensile strength and time of fracture for the coated Mg-Zn-Ca alloy are higher than those of the uncoated one, which is beneficial in supporting fractured bone for a longer time. Thus Mg-Zn-Ca alloy coated with Ca-def HA is be a promising candidate for biodegradable orthopaedic implants, and is worthwhile to further investigate the in vivo degradation behavior. PMID- 20004747 TI - Polymeric micelle-templated synthesis of hydroxyapatite hollow nanoparticles for a drug delivery system. AB - Hydroxyapatite (HA) hollow nanoparticles (HNPs) have great potential in nanoscaled delivery devices due to their small size, excellent biocompatibility and expected high capacity. However, the preparation of HA HNPs for their application in a drug delivery system has rarely been reported because HA has a complicated crystal structure and it is difficult to obtain stable HA HNPs with hollows that are only nanoscaled in size. In the present study, HA HNPs were successfully produced through a novel polymeric micelle-templating method. The micelles were structured with completely insoluble Pluronic P123 molecules at cloud point as the core and Tween-60 molecules as the shell by the hydrophobic interaction of the alkyl chains with the insoluble P123 core. The morphology of the HA HNPs could be transformed from nanospheres to nanotubes by adding citric acid as a cosurfactant. The prepared HA HNPs had a much higher drug payload than traditional nanoparticles, using vancomycin as the model drug. Most importantly, the HA nanotubes were coupled with a layer of citrate molecules on the HA surfaces, which could further improve the drug load efficiency and could form an excellent pH-controlled open/closed gate for drug release with the addition of cationic polyelectrolytes. PMID- 20004748 TI - In vitro and in vivo evaluation of a novel nanosize hydroxyapatite particles/poly(ester-urethane) composite scaffold for bone tissue engineering. AB - Scaffolds for bone tissue engineering should provide an osteoconductive surface to promote the ingrowth of new bone after implantation into bone defects. This may be achieved by hydroxyapatite loading of distinct scaffold biomaterials. Herein, we analyzed the in vitro and in vivo properties of a novel nanosize hydroxyapatite particles/poly(ester-urethane) (nHA/PU) composite scaffold which was prepared by a salt leaching-phase inverse process. Microtomography, scanning electron microscopy and X-ray spectroscopy analyses demonstrated the capability of the material processing to create a three-dimensional porous PU scaffold with nHA on the surface. Compared to nHA-free PU scaffolds (control), this modified scaffold type induced a significant increase in in vitro adsorption of model proteins. In vivo analysis of the inflammatory and angiogenic host tissue response to implanted nHA/PU scaffolds in the dorsal skinfold chamber model indicated that the incorporation of nHA particles into the scaffold material did not affect biocompatibility and vascularization when compared to control scaffolds. Thus, nHA/PU composite scaffolds represent a promising new type of scaffold for bone tissue engineering, combining the flexible material properties of PU with the advantage of an osteoconductive surface. PMID- 20004749 TI - Degradation studies on segmented polyurethanes prepared with HMDI, PCL and different chain extenders. AB - Biodegradable segmented polyurethanes (BSPUs) were prepared with poly(caprolactone) as a soft segment, 4,4'-methylene bis (cyclohexyl isocyanate) and either butanediol (BSPU1) or dithioerythritol (BSPU2) as a chain extender. BSPU samples were characterized in terms of their physicochemical properties and their hemocompatibility. Polymers were then degraded in acidic (HCl 2N), alkaline (NaOH 5M) and oxidative (H(2)O(2) 30wt.%) media and characterized by their mass loss, Fourier transform infrared (FTIR), differential scanning calorimetry (DSC), thermogravimetric analysis (TGA), X-ray diffraction (XRD) and scanning electron microscopy (SEM). Undegraded BSPU1 and BSPU2 exhibited different properties, such as the glass transition temperature T(g) of the soft segment (-25 vs. 4 degrees C), mechanical properties (600% vs. 900% strain to break) and blood coagulating properties (clotting time=11.46 vs. 8.13min). After acidic and alkaline degradation, the disappearance of the 1728cm(-1) band of polycaprolactone (PCL) on both types of BSPU was detected by FTIR. However, the oxidative environment did not affect the soft segment severely as the presence of PCL crystalline domains were observed both by DSC (melting temperature T(m)=52.8 degrees C) and XRD (2theta=21.3 degrees and 23.7 degrees ). By TGA three decomposition temperatures were recorded for both BSPU samples, but the higher decomposition temperature was enhanced after acidic and alkaline degradation. The formation of the porous structure on BSPU1 was observed by SEM, while a granular surface was observed on BSPU2 after alkaline degradation. PMID- 20004750 TI - A new approach in biomimetic synthesis of calcium phosphate coatings using lactic acid-Na lactate buffered body fluid solution. AB - The main objective of this study was to investigate calcium phosphate (CaP) coatings on Ti6Al4V substrates by using the biomimetic technique. To this purpose, a new solution was developed to coat CaP on Ti6Al4V alloy substrates. The newly formulated body fluid (Lac-SBF) contained appropriate amounts of sodium lactate (NaL) and lactic acid (HL), as well as all the other ionic constituents of the human blood plasma. The inorganic ion concentrations of the Lac-SBF solutions were identical with those of human blood plasma. The new Lac-SBF solution of this study eliminated the need for using Tris/HCl or Hepes/NaOH buffers. Prior to coating, Ti6Al4V substrates were chemically treated in NaOH and/or NaOH+H(2)O(2) solutions as an alternative route and then heated at 600 degrees C for 1h in air. In the previous applications, the Cl(-) ion concentration was found to be higher than blood plasma 103mM, which exists in human blood plasma as a result of Tris/HCl which are used to prevent precipitation and to keep the pH level at certain values. In this study, instead of using Tris/HCl, HL/NaL which are generated by human body and do not show any toxic behavior, are used and Cl(-) concentration was kept at 103mM value for the first time. The prepared Lac-SBF was shown to have similar concentration to human blood plasma in terms of all inorganic ions for the first time. Solution properties were evaluated by using turbidimeter, pH meter and rheometer. The coatings were characterized using Fourier-transform infrared spectroscopy (FTIR), X-ray diffraction (XRD), scanning electron microscopy (SEM), and a scratch tester. The obtained results are presented and discussed. PMID- 20004751 TI - Mechanical properties and osteocompatibility of novel biodegradable alanine based polyphosphazenes: Side group effects. AB - The versatility of polymers for tissue regeneration lies in the feasibility to modulate the physical and biological properties by varying the side groups grafted to the polymers. Biodegradable polyphosphazenes are high-molecular-weight polymers with alternating nitrogen and phosphorus atoms in the backbone. This study is the first of its kind to systematically investigate the effect of side group structure on the compressive strength of novel biodegradable polyphosphazene based polymers as potential materials for tissue regeneration. The alanine polyphosphazene based polymers, poly(bis(ethyl alanato) phosphazene) (PNEA), poly((50% ethyl alanato) (50% methyl phenoxy) phosphazene) (PNEA(50)mPh(50)), poly((50% ethyl alanato) (50% phenyl phenoxy) phosphazene) (PNEA(50)PhPh(50)) were investigated to demonstrate their mechanical properties and osteocompatibility. Results of mechanical testing studies demonstrated that the nature and the ratio of the pendent groups attached to the polymer backbone play a significant role in determining the mechanical properties of the resulting polymer. The compressive strength of PNEA(50)PhPh(50) was significantly higher than poly(lactide-co-glycolide) (85:15 PLAGA) (p<0.05). Additional studies evaluated the cellular response and gene expression of primary rat osteoblast cells on PNEA, PNEA(50)mPh(50) and PNEA(50)PhPh(50) films as candidates for bone tissue engineering applications. Results of the in vitro osteocompatibility evaluation demonstrated that cells adhere, proliferate, and maintain their phenotype when seeded directly on the surface of PNEA, PNEA(50)mPh(50), and PNEA(50)PhPh(50). Moreover, cells on the surface of the polymers expressed type I collagen, alkaline phosphatase, osteocalcin, osteopontin, and bone sialoprotein, which are characteristic genes for osteoblast maturation, differentiation, and mineralization. PMID- 20004753 TI - MBD3 mutations are not responsible for ICR1 hypomethylation in Silver-Russell syndrome. AB - Silver-Russell syndrome (SRS) is a sporadic and heterogeneous disease that is mainly associated with intrauterine and postnatal growth retardation. The most frequent known aberration in SRS patients is a hypomethylation of the imprinting control region 1 (ICR1) in 11p15 ( approximately 38%). Up to now the basic mechanisms leading to this imprinting defect are unknown. Based on the recent findings that a reduced level of the methyl-CpG binding protein 3 (Mbd3) in mice results in a specific hypomethylation of the ICR1 and in a smaller size of embryos we hypothesized that mutations in the genomic sequence of the human MBD3 gene might cause SRS. We carried out mutation analysis of MBD3 in 20 SRS patients with hypomethylation of the ICR1 but did not detect any pathogenic variant in the coding region. Thus we assume that genomic mutations of MBD3 are not relevant for the aetiology of the ICR1 hypomethylation and therefore for SRS. PMID- 20004752 TI - Prenatal study of common submicroscopic "genomic disorders" using MLPA with subtelomeric/microdeletion syndrome probe mixes, among gestations with ultrasound abnormalities in the first trimester. AB - PURPOSE: The present study aims to investigate the presence of common submicroscopic chromosomal rearrangements in fetuses with ultrasound abnormalities or positive screening in the first trimester and normal karyotype. We used the multiplex ligation-dependent probe amplification (MLPA) technique with subtelomeric (SALSA P036B) and microdeletion syndrome (SALSA P064B/P096) probe mixes as a screening method to measure copy number changes on the tested probes in chorionic villus sampling. MLPA with P036B and P064/P096 probe mixes was performed on 49 chorionic villi DNA samples obtained between the 11th and 13th week of gestation. RESULTS: The MLPA analyses did not detect any diminished or increased intensity for all the tested probes in the samples. CONCLUSIONS: Our results suggest that the common submicroscopic "genomic disorders" (microdeletion and microduplication syndromes) would not be frequently detected in the first trimester anomalies screening. PMID- 20004754 TI - Immobilization of Candida antarctica lipase B by adsorption in organic medium. AB - Candida antarctica lipase B (CALB) was immobilized on the macroporous resin by physical adsorption in organic medium. The immobilization was performed in 5 mL isooctane, and the immobilization conditions were optimized. The results were achieved with the mass ratio of lipase to support 1:80, the buffer of pH 6.0, initial addition of PBS 75 microL, and immobilization time of two hours at 30 degrees C. Under the optimal conditions, the activity recovery was 83.3%. IM-CALB presented enhanced pH and thermal stability compared to the free lipase, and showed comparable stability with the commercial Novozym 435, after 7 times repeated use for catalyzing the synthesis of ethyl lactate, 56.9% of its initial activity was retained, and only 24.7% was retained when used for catalyzing the hydrolysis of olive oil. PMID- 20004755 TI - 2D-electrophoresis and the urine proteome map: where do we stand? AB - The discovery of urinary biomarkers is a main topic in clinical medicine. The development of proteomics has rapidly changed the knowledge on urine protein composition and probably will modify it again. Two-dimensional electrophoresis (2D-PAGE) coupled with mass spectrometry has represented for years the technique of choice for the analysis of urine proteins and it is time to draw some conclusions. This review will focus on major methodological aspects related to urine sample collection, storage and analysis by 2D-PAGE and attempt to define an advanced normal urine protein map. Overall, 1118 spots were reproducibly found in normal urine samples but only 275 were characterized as isoforms of 82 proteins. One-hundred height spots belonging to 30 proteins were also detected in plasma and corresponded to typical plasma components. The identity of most of the proteins found in normal urine by 2D-PAGE remains to be determined, the majority being low-molecular weight proteins (<30 kDa). Equalization procedures would also enhance sensitivity of the analysis and allow low abundance proteins to be characterized. Therefore, we are still on the way to define the normal urine composition. Technology advancements in concentrating procedure will improve sensitivity and give the possibility to purify proteins for mass spectrometry. PMID- 20004756 TI - Genetic architecture of hand quantitative ultrasound measures: a population-based study in a Sardinian genetic isolate. AB - It is now recognized that quantitative ultrasound (QUS) measures may predict osteoporotic fracture risk independently of bone mineral density. Although many studies have examined genetic and environmental components of bone mineral density and calcaneal QUS measures, few of them were addressed to phalangeal QUS phenotypes, and none to graphic trace parameters. This study aims to evaluate the relative contribution of genetics in the expression of phalangeal QUS traits in the adult healthy population of a Sardinian genetic isolate. Our sample includes 6056 men and women aged 30-103 years, from 43 extended pedigrees recruited in 10 villages of Ogliastra region in occasion of a large epidemiologic survey. Amplitude-dependent speed of sound (AD-SoS), fast wave amplitude (FWA), signal dynamic (SDy), bone transmission time (BTT) and ultrasound bone profile index (UBPI) were obtained from the non-dominant hand using the IGEA DBM Sonic Bone Profiler. These phenotypes were first regressed on age, anthropometric and bioimpedance measures, serum calcium, phosphorus and alkaline phosphatase, alcohol and caffeine consumption, smoking status, exercise and also months since menopause and estrogens use in women. Adjusted QUS parameters were then analyzed by univariate and bivariate variance component models to obtain heritability estimates and genetic and environmental correlations. QUS parameters were correlated to age, anthropometric and bioimpedance measures, serum phosphorus, alkaline phosphatase and to reproductive history and menopause in women. All phenotypes demonstrated substantial heritabilities ranging from 0.29+/-0.03 for SDy to 0.55+/-0.03 for FWA. Proportion of variance due to all covariates ranged from 36% for SDy to 59% for BTT. Many significant genetic and environmental correlations were found between the different QUS measures. In this study, genetic factors appear to play a relevant role in determining hand QUS measures even when taking into account various important environmental factors. Furthermore, the modest genetic correlations may imply the existence of partially unique sets of genes affecting different QUS traits, thus suggesting that QUS parameters measure different properties of bone tissue. PMID- 20004758 TI - Optimum frequency of exercise for bone health: randomised controlled trial of a high-impact unilateral intervention. AB - INTRODUCTION: Exercise can increase bone strength, but to be effective in reducing fracture risk, exercise must be feasible enough to be adopted into daily life and influence potentially vulnerable skeletal sites such as the superolateral cortex of the femoral neck, where thinning is associated with increased fracture risk. Brief, high-impact exercise increases femoral neck bone density but the optimal frequency of such exercise and the location of bone accrual is unknown. This study thus examined (1) the effectiveness of different weekly frequencies of exercise on femoral neck BMD and (2) whether BMD change differed between hip sites using a high-impact, unilateral intervention. METHODS: Healthy premenopausal women were randomly assigned to exercise 0, 2, 4, or 7 days/week for 6 months. The exercise intervention incorporated 50 multidirectional hops on one randomly selected leg. BMD was measured by DXA at baseline and after 6 months of exercise. Changes in the exercise leg were compared between groups using ANCOVA, with change in the control leg and baseline BMD as covariates. RM-MANOVA was conducted to determine whether bone changes from exercise differed between hip sites. RESULTS: 61 women (age 33.6+/-11.1 years) completed the intervention. Compliance amongst exercisers was 86.7+/-10.6%. Peak ground reaction forces during exercise increased from 2.5 to 2.8 times body weight. The change in femoral neck BMD in the exercise limb (adjusted for change in the control limb and baseline BMD) differed between groups (p=0.015), being 0.3% (-1.2 to 0.6), 0.0% (-1.0 to 1.0), 0.9% (-0.1 to 2.0) and 1.8% (0.8 to 2.8) in those exercising 0, 2, 4 and 7 days per week, respectively. When BMD changes at upper neck, lower neck and trochanter were compared using RM-MANOVA, a significant exercise effect was observed (p=0.048), but this did not differ significantly between sites (p=0.439) despite greatest mean increases at the upper femoral neck. CONCLUSIONS: Brief, daily hopping exercises increased femoral neck BMD in premenopausal women but less frequent exercise was not effective. Brief high-impact exercise may have a role in reducing hip fragility, but may need to be performed frequently for optimal response. PMID- 20004757 TI - The cystic fibrosis transmembrane conductance regulator (CFTR) is expressed in maturation stage ameloblasts, odontoblasts and bone cells. AB - Patients with cystic fibrosis (CF) have mild defects in dental enamel. The gene mutated in these patients is CFTR, a Cl(-) channel involved in transepithelial salt and water transport and bicarbonate secretion. We tested the hypothesis that Cftr channels are present and operating in the plasma membranes of mouse ameloblasts. Tissue sections of young mouse jaws and fetal human jaws were immunostained with various anti-Cftr antibodies. Specificity of the antibodies was validated in Cftr-deficient murine and human tissues. Immunostaining for Cftr was obtained in the apical plasma membranes of mouse maturation ameloblasts of both incisor and molar tooth germs. A granular intracellular immunostaining of variable intensity was also noted in bone cells and odontoblasts. In Cftr deficient mice the incisors were chalky white and eroded much faster than in wild type mice. Histologically, only maturation ameloblasts of incisors were structurally affected in Cftr-deficient mice. Some antibody species gave also a positive cytosolic staining in Cftr-deficient cells. Transcripts of Cftr were found in maturation ameloblasts, odontoblasts and bone cells. Similar data were obtained in forming human dentin and bone. We conclude that Cftr protein locates in the apical plasma membranes of mouse maturation ameloblasts. In mouse incisors Cftr is critical for completion of enamel mineralization and conceivably functions as a regulator of pH during rapid crystal growth. Osteopenia found in CF patients as well as in Cftr-deficient mice is likely associated with defective Cftr operating in bone cells. PMID- 20004759 TI - The biological coherence of human phenome databases. AB - Disease networks are increasingly explored as a complement to networks centered around interactions between genes and proteins. The quality of disease networks is heavily dependent on the amount and quality of phenotype information in phenotype databases of human genetic diseases. We explored which aspects of phenotype database architecture and content best reflect the underlying biology of disease. We used the OMIM-based HPO, Orphanet, and POSSUM phenotype databases for this purpose and devised a biological coherence score based on the sharing of gene ontology annotation to investigate the degree to which phenotype similarity in these databases reflects related pathobiology. Our analyses support the notion that a fine-grained phenotype ontology enhances the accuracy of phenome representation. In addition, we find that the OMIM database that is most used by the human genetics community is heavily underannotated. We show that this problem can easily be overcome by simply adding data available in the POSSUM database to improve OMIM phenotype representations in the HPO. Also, we find that the use of feature frequency estimates--currently implemented only in the Orphanet database- significantly improves the quality of the phenome representation. Our data suggest that there is much to be gained by improving human phenome databases and that some of the measures needed to achieve this are relatively easy to implement. More generally, we propose that curation and more systematic annotation of human phenome databases can greatly improve the power of the phenotype for genetic disease analysis. PMID- 20004760 TI - Dosage-dependent severity of the phenotype in patients with mental retardation due to a recurrent copy-number gain at Xq28 mediated by an unusual recombination. AB - We report on the identification of a 0.3 Mb inherited recurrent but variable copy number gain at Xq28 in affected males of four unrelated families with X-linked mental retardation (MR). All aberrations segregate with the disease in the families, and the carrier mothers show nonrandom X chromosome inactivation. Tiling Xq28-region-specific oligo array revealed that all aberrations start at the beginning of the low copy repeat LCR-K1, at position 153.20 Mb, and end just distal to LCR-L2, at 153.54 Mb. The copy-number gain always includes 18 annotated genes, of which RPL10, ATP6AP1 and GDI1 are highly expressed in brain. From these, GDI1 is the most likely candidate gene. Its copy number correlates with the severity of clinical features, because it is duplicated in one family with nonsyndromic moderate MR, is triplicated in males from two families with mild MR and additional features, and is present in five copies in a fourth family with a severe syndromic form of MR. Moreover, expression analysis revealed copy-number dependent increased mRNA levels in affected patients compared to control individuals. Interestingly, analysis of the breakpoint regions suggests a recombination mechanism that involves two adjacent but different sets of low copy repeats. Taken together, our data strongly suggest that an increased expression of GDI1 results in impaired cognition in a dosage-dependent manner. Moreover, these data also imply that a copy-number gain of an individual gene present in the larger genomic aberration that leads to the severe MECP2 duplication syndrome can of itself result in a clinical phenotype as well. PMID- 20004761 TI - Meta-analysis of genome-wide association studies with overlapping subjects. AB - Data from multiple genome-wide association studies are often analyzed together for the purposes of combining information from several studies of the same disease or comparing results across different disorders. We provide a valid and efficient approach to such meta-analysis, allowing for overlapping study subjects. The available data may contain individual participant records or only meta-analytic summary results. Simulation studies demonstrate that failure to account for overlapping subjects can greatly inflate type I error when combining results from multiple studies of the same disease and can drastically reduce power when comparing results across different disorders. In addition, the proposed approach can be substantially more powerful than the simple approach of splitting the overlapping subjects among studies, especially for comparing results across different disorders. The advantages of the new approach are illustrated with empirical data from two sets of genome-wide association studies. PMID- 20004762 TI - Loss of dermatan-4-sulfotransferase 1 function results in adducted thumb-clubfoot syndrome. AB - Adducted thumb-clubfoot syndrome is an autosomal-recessive disorder characterized by typical facial appearance, wasted build, thin and translucent skin, congenital contractures of thumbs and feet, joint instability, facial clefting, and coagulopathy, as well as heart, kidney, or intestinal defects. We elucidated the molecular basis of the disease by using a SNP array-based genome-wide linkage approach that identified distinct homozygous nonsense and missense mutations in CHST14 in each of four consanguineous families with this disease. The CHST14 gene encodes N-acetylgalactosamine 4-O-sulfotransferase 1 (D4ST1), which catalyzes 4-O sulfation of N-acetylgalactosamine in the repeating iduronic acid-alpha1,3-N acetylgalactosamine disaccharide sequence to form dermatan sulfate. Mass spectrometry of glycosaminoglycans from a patient's fibroblasts revealed absence of dermatan sulfate and excess of chondroitin sulfate, showing that 4-O sulfation by CHST14 is essential for dermatan sulfate formation in vivo. Our results indicate that adducted thumb-clubfoot syndrome is a disorder resulting from a defect specific to dermatan sulfate biosynthesis and emphasize roles for dermatan sulfate in human development and extracellular-matrix maintenance. PMID- 20004763 TI - A truncating mutation of TRAPPC9 is associated with autosomal-recessive intellectual disability and postnatal microcephaly. AB - Although autosomal genes are increasingly recognized as important causes of intellectual disability, very few of them are known. We identified a genetic locus for autosomal-recessive nonsyndromic intellectual disability associated with variable postnatal microcephaly through homozygosity mapping of a consanguineous Israeli Arab family. Sequence analysis of genes in the candidate interval identified a nonsense nucleotide change in the gene that encodes TRAPPC9 (trafficking protein particle complex 9, also known as NIBP), which has been implicated in NF-kappaB activation and possibly in intracellular protein trafficking. TRAPPC9 is highly expressed in the postmitotic neurons of the cerebral cortex, and MRI analysis of affected patients shows defects in axonal connectivity. This suggests essential roles of TRAPPC9 in human brain development, possibly through its effect on NF-kappaB activation and protein trafficking in the postmitotic neurons of the cerebral cortex. PMID- 20004764 TI - Combination of linkage mapping and microarray-expression analysis identifies NF kappaB signaling defect as a cause of autosomal-recessive mental retardation. AB - Autosomal-recessive inheritance accounts for nearly 25% of nonsyndromic mental retardation (MR), but the extreme heterogeneity of such conditions markedly hampers gene identification. Combining autozygosity mapping and RNA expression profiling in a consanguineous Tunisian family of three MR children with mild microcephaly and white-matter abnormalities identified the TRAPPC9 gene, which encodes a NF-kappaB-inducing kinase (NIK) and IkappaB kinase complex beta (IKK beta) binding protein, as a likely candidate. Sequencing analysis revealed a nonsense variant (c.1708C>T [p.R570X]) within exon 9 of this gene that is responsible for an undetectable level of TRAPPC9 protein in patient skin fibroblasts. Moreover, TNF-alpha stimulation assays showed a defect in IkBalpha degradation, suggesting impaired NF-kappaB signaling in patient cells. This study provides evidence of an NF-kappaB signaling defect in isolated MR. PMID- 20004765 TI - Identification of mutations in TRAPPC9, which encodes the NIK- and IKK-beta binding protein, in nonsyndromic autosomal-recessive mental retardation. AB - Mental retardation/intellectual disability is a devastating neurodevelopmental disorder with serious impact on affected individuals and their families, as well as on health and social services. It occurs with a prevalence of approximately 2%, is an etiologically heterogeneous condition, and is frequently the result of genetic aberrations. Autosomal-recessive forms of nonsyndromic MR (NS-ARMR) are believed to be common, yet only five genes have been identified. We have used homozygosity mapping to search for the gene responsible for NS-ARMR in a large Pakistani pedigree. Using Affymetrix 5.0 single nucleotide polymorphism (SNP) microarrays, we identified a 3.2 Mb region on 8q24 with a continuous run of 606 homozygous SNPs shared among all affected members of the family. Additional genotype data from microsatellite markers verified this, allowing us to calculate a two-point LOD score of 5.18. Within this region, we identified a truncating homozygous mutation, R475X, in exon 7 of the gene TRAPPC9. In a second large NS ARMR/ID family, previously linked to 8q24 in a study of Iranian families, we identified a 4 bp deletion within exon 14 of TRAPPC9, also segregating with the phenotype and truncating the protein. This gene encodes NIK- and IKK-beta-binding protein (NIBP), which is involved in the NF-kappaB signaling pathway and directly interacts with IKK-beta and MAP3K14. Brain magnetic resonance imaging of affected individuals indicates the presence of mild cerebral white matter hypoplasia. Microcephaly is present in some but not all affected individuals. Thus, to our knowledge, this is the sixth gene for NS-ARMR to be discovered. PMID- 20004766 TI - Homozygous inactivating mutations in the NKX3-2 gene result in spondylo megaepiphyseal-metaphyseal dysplasia. AB - Spondylo-megaepiphyseal-metaphyseal dysplasia (SMMD) is a rare skeletal dysplasia with only a few cases reported in the literature. Affected individuals have a disproportionate short stature with a short and stiff neck and trunk. The limbs appear relatively long and may show flexion contractures of the distal joints. The most remarkable radiographic features are the delayed and impaired ossification of the vertebral bodies as well as the presence of large epiphyseal ossification centers and wide growth plates in the long tubular bones. Numerous pseudoepiphyses of the short tubular bones in hands and feet are another remarkable feature of the disorder. Genome wide homozygosity mapping followed by a candidate gene approach resulted in the elucidation of the genetic cause in three new consanguineous families with SMMD. Each proband was homozygous for a different inactivating mutation in NKX3-2, a homeobox-containing gene located on chromosome 4p15.33. Striking similarities were found when comparing the vertebral ossification defects in SMMD patients with those observed in the Nkx3-2 null mice. Distinguishing features were the asplenia found in the mutant mice and the radiographic abnormalities in the limbs only observed in SMMD patients. The absence of the latter anomalies in the murine model may be due to the perinatal death of the affected animals. This study illustrates that NKX3-2 plays an important role in endochondral ossification of both the axial and appendicular skeleton in humans. In addition, it defines SMMD as yet another skeletal dysplasia with autosomal-recessive inheritance and a distinct phenotype. PMID- 20004767 TI - Remarkably little variation in proteins encoded by the Y chromosome's single-copy genes, implying effective purifying selection. AB - Y-linked single-nucleotide polymorphisms (SNPs) have served as powerful tools for reconstructing the worldwide genealogy of human Y chromosomes and for illuminating patrilineal relationships among modern human populations. However, there has been no systematic, worldwide survey of sequence variation within the protein-coding genes of the Y chromosome. Here we report and analyze coding sequence variation among the 16 single-copy "X-degenerate" genes of the Y chromosome. We examined variation in these genes in 105 men representing worldwide diversity, resequencing in each man an average of 27 kb of coding DNA, 40 kb of intronic DNA, and, for comparison, 15 kb of DNA in single-copy Y chromosomal pseudogenes. There is remarkably little variation in X-degenerate protein sequences: two chromosomes drawn at random differ on average by a single amino acid, with half of these differences arising from a single, conservative Asp-->Glu mutation that occurred approximately 50,000 years ago. Further analysis showed that nucleotide diversity and the proportion of variant sites are significantly lower for nonsynonymous sites than for synonymous sites, introns, or pseudogenes. These differences imply that natural selection has operated effectively in preserving the amino acid sequences of the Y chromosome's X degenerate proteins during the last approximately 100,000 years of human history. Thus our findings are at odds with prominent accounts of the human Y chromosome's imminent demise. PMID- 20004768 TI - mtDNA data mining in GenBank needs surveying. PMID- 20004770 TI - Genetic landscape of Eurasia and "admixture" in Uyghurs. PMID- 20004772 TI - Haplotype background, repeat length evolution, and Huntington's disease. PMID- 20004773 TI - Response to Falush: a role for cis-element polymorphisms in HD. PMID- 20004774 TI - Therapy of autoinflammatory syndromes. AB - The therapy of autoinflammatory syndromes is an excellent example of the power of translational research. Recent advances in our understanding of the molecular and immunologic basis of this newly identified classification of disease have allowed for the application of novel, effective, targeted treatments with life-changing effects on patients. Although colchicine and TNF-alpha inhibitors are important therapies for specific autoinflammatory syndromes, the novel IL-1-targeted drugs are particularly effective for many of these diseases. Recently, the pharmaceutical industry has adopted a strategy of confirming the efficacy of new targeted drugs in often-ignored patients with orphan diseases, and US Food and Drug Administration policies have allowed for accelerated approval of these drugs, creating a win-win situation for patients and industry. This article reviews the general approach to the therapy of autoinflammatory diseases, focusing on current approved therapies and novel approaches that might be used in the future. PMID- 20004775 TI - Autoinflammation: the prominent role of IL-1 in monogenic autoinflammatory diseases and implications for common illnesses. AB - The discovery of the genetic causes of a rare group of immune-mediated inflammatory conditions that mimic infections and allergic conditions in their clinical presentation and the molecular understanding of the function of the mutated molecules in these diseases has led to a revolution in our understanding of the pathogenesis of systemic and local inflammation. The proteins mutated in a number of these so-called autoinflammatory diseases are part of, or regulate the activity of, intracellular molecular complexes, the inflammasomes, that sense "danger" to the body and coordinate an initial immune response. Our understanding of specific triggers of the inflammasomes, coupled with the recognition that inflammasomes are critical for activation of the proinflammatory cytokine IL-1, has provided a rational and very effective target in the treatment of a number of these rare autoinflammatory diseases. In addition, the ongoing discovery of the role of inflammasomes and IL-1 activation and secretion in a number of genetically complex disorders have fundamentally changed our view of disease pathogenesis in a growing number of disorders that were heretofore not even thought of as "immunologic" diseases. PMID- 20004777 TI - Primary immunodeficiencies: 2009 update. AB - More than 50 years after Ogdeon Bruton's discovery of congenital agammaglobulinemia, human primary immunodeficiencies (PIDs) continue to unravel novel molecular and cellular mechanisms that govern development and function of the human immune system. This report provides the updated classification of PIDs that has been compiled by the International Union of Immunological Societies Expert Committee on Primary Immunodeficiencies after its biannual meeting in Dublin, Ireland, in June 2009. Since the appearance of the last classification in 2007, novel forms of PID have been discovered, and additional pathophysiology mechanisms that account for PID in human beings have been unraveled. Careful analysis and prompt recognition of these disorders is essential to prompt effective forms of treatment and thus to improve survival and quality of life in patients affected with PIDs. PMID- 20004779 TI - Thirteen-year follow-up of early intervention with an inhaled corticosteroid in patients with asthma. AB - BACKGROUND: In a 3-year study, adult patients who recently developed asthma (symptoms for less than 1 year) were treated for 2 years with the inhaled corticosteroid (ICS) budesonide (early therapy) or terbutaline. During the third year of the study, terbutaline-treated patients received budesonide (delayed therapy). Differences in lung function and bronchial responsiveness to histamine were observed between the 2 groups. OBJECTIVE: We compared the effects of early versus delayed budesonide therapy after a 10-year follow-up period (13 years after the study began) and current real-life data. METHODS: Of the original 103 patients, 90 were re-examined 13 years after study initiation. After the third year of the study, all patients had their medications, including the dose of ICS, individually adjusted. RESULTS: After the follow-up period, lung function was within the normal range for the entire group (all patients); bronchial responsiveness significantly improved compared with baseline data. No statistically significant differences in clinical or functional variables were found between patients given early or delayed budesonide therapy. However, the delayed therapy group had a higher neutrophil count and higher concentrations of eosinophilic cationic protein and myeloperoxidase in induced sputum. This group had also used more asthma medication and hospital days. CONCLUSIONS: Patients with relatively mild asthma who received ICS within 12 months of their first asthma symptoms or after a 2-year delay achieved equally good functional control of asthma after 10 years of individualized therapy. However, the delayed therapy group exhibited slightly less optimal disease control and more signs of airway inflammation. PMID- 20004780 TI - New-onset mild asthma III: a sequel to 2 "Oscar winners". PMID- 20004776 TI - Improving cellular therapy for primary immune deficiency diseases: recognition, diagnosis, and management. AB - More than 20 North American academic centers account for the majority of hematopoietic stem cell transplantation (HCT) procedures for primary immunodeficiency diseases (PIDs), with smaller numbers performed at additional sites. Given the importance of a timely diagnosis of these rare diseases and the diversity of practice sites, there is a need for guidance as to best practices in management of patients with PIDs before, during, and in follow-up for definitive treatment. In this conference report of immune deficiency experts and HCT physicians who care for patients with PIDs, we present expert guidance for (1) PID diagnoses that are indications for HCT, including severe combined immunodeficiency disease (SCID), combined immunodeficiency disease, and other non SCID diseases; (2) the critical importance of a high degree of suspicion of the primary care physician and timeliness of diagnosis for PIDs; (3) the need for rapid referral to an immune deficiency expert, center with experience in HCT, or both for patients with PIDs; (4) medical management of a child with suspicion of SCID/combined immunodeficiency disease while confirming the diagnosis, including infectious disease management and workup; (5) the posttransplantation follow-up visit schedule; (6) antimicrobial prophylaxis after transplantation, including gamma globulin administration; and (7) important indications for return to the transplantation center after discharge. Finally, we discuss the role of high quality databases in treatment of PIDs and HCT as an element of the infrastructure that will be needed for productive multicenter clinical trials in these rare diseases. PMID- 20004781 TI - Prolonged bronchoprotection against inhaled methacholine by inhaled BI 1744, a long-acting beta(2)-agonist, in patients with mild asthma. AB - BACKGROUND: Long-acting ss(2)-agonists are an established controller medication in asthma. BI 1744 is a novel L?long-acting ss(2)-agonist with a preclinical profile that suggests 24-hour bronchodilation and bronchoprotection may be achieved. OBJECTIVE: To examine the bronchoprotective effects of single doses of BI 1744 against methacholine provocation in subjects with mild asthma. METHODS: Thirty-one subjects with mild asthma were randomized to receive single doses of BI 1744 (2, 5, 10, 20 microg) or placebo on separate days according to a double blind, 5-way crossover design. Methacholine challenges were performed at 30 minutes and at 4, 8, 24, and 32 hours after each single dose of medication, and the results were expressed as PC(20) FEV(1). RESULTS: All doses of BI 1744 produced statistically significant increases in the methacholine PC(20) compared with placebo as long as 32 hours. The mean (geometric SEM) methacholine PC(20) 24 hours after dosing with placebo was 1.73 (1.13) mg/mL, which increased after 2 microg to 3.86 (1.14) mg/mL, after 5 microg to 5.67 (1.14) mg/mL, after 10 microg to 9.42 (1.13) mg/mL, and after 20 microg to 13.71 (1.14) mg/mL (all P < .0001). After 32 hours, the methacholine PC(20) value remained significantly increased for all doses. No safety or tolerability concerns were identified. CONCLUSION: BI 1744 provides significant bronchoprotection against inhaled methacholine for up to 32 hours after single-dose administration. PMID- 20004782 TI - Broad defects in epidermal cornification in atopic dermatitis identified through genomic analysis. AB - BACKGROUND: Psoriasis and atopic dermatitis (AD) are common, complex inflammatory skin diseases. Both diseases display immune infiltrates in lesions and epidermal growth/differentiation alterations associated with a defective skin barrier. An incomplete understanding of differences between these diseases makes it difficult to compare human disease pathology to animal disease models. OBJECTIVE: To characterize differences between these diseases in expression of genes related to epidermal growth/differentiation and inflammatory circuits. METHODS: We performed genomic profiling of mRNA in chronic psoriasis (n = 15) and AD (n = 18) skin lesions compared with normal human skin (n = 15). RESULTS: As expected, clear disease classifications could be constructed on the basis of expected immune polarity (T(H)1, T(H)2, T(H)17) differences. However, even more striking differences were identified in epidermal differentiation programs that could be used for precise disease classifications. Although both psoriasis and AD skin lesions displayed regenerative epidermal hyperplasia, which is a general alteration in epidermal growth, keratinocyte terminal differentiation was differentially polarized. In AD, we found selective defects in expression of multiple genes encoding the cornified envelope, with the largest alteration in loricrin (expressed at 2% of the level of normal skin). At the ultrastructural level, the cornified envelope in AD was broadly defective with highly decreased compaction of corneocytes and reduced intercellular lipids. Hence, the entire keratinocyte terminal differentiation program (cytoplasmic compaction, cornification, and lipid release) is defective in AD, potentially underlying the immune differences. CONCLUSION: Our study shows that although alterations in barrier responses exist in both diseases, epidermal differentiation is differentially polarized, with major implications for primary disease pathogenesis. PMID- 20004784 TI - Epinephrine treatment is infrequent and biphasic reactions are rare in food induced reactions during oral food challenges in children. AB - BACKGROUND: Data about epinephrine use and biphasic reactions in childhood food induced anaphylaxis during oral food challenges are scarce. OBJECTIVE: To determine the prevalence and risk factors of reactions requiring epinephrine and the rate of biphasic reactions during oral food challenges (OFCs) in children. METHODS: Reaction details of positive OFCs in children between 1999 and 2007 were collected by using a computerized database. Selection of patients for OFCs was generally predicated on < or =50% likelihood of a positive challenge and a low likelihood of a severe reaction on the basis of the clinical history, specific IgE levels, and skin prick tests. RESULTS: A total of 436 of 1273 OFCs resulted in a reaction (34%). Epinephrine was administered in 50 challenges (11% of positive challenges, 3.9% overall) for egg (n = 15, 16% of positive OFCs to egg), milk (n = 14, 12%), peanut (n = 10, 26%), tree nuts (n = 4, 33%), soy (n = 3, 7%), wheat (n = 3, 9%), and fish (n = 1, 9%). Reactions requiring epinephrine occurred in older children (median, 7.9 vs 5.8 years; P < .001) and were more often caused by peanuts (P = .006) compared with reactions not treated with epinephrine. There was no difference in the sex, prevalence of asthma, history of anaphylaxis, specific IgE level, skin prick tests, or amount of food administered. Two doses of epinephrine were required in 3 of 50 patients (6%) reacting to wheat, cow's milk, and pistachio. There was 1 (2%) biphasic reaction. No reaction resulted in life-threatening respiratory or cardiovascular compromise. CONCLUSION: Older age and reactions to peanuts were risk factors for anaphylaxis during oral food challenges. Reactions requiring multiple doses of epinephrine and biphasic reactions were infrequent. PMID- 20004785 TI - Large deletions and point mutations involving the dedicator of cytokinesis 8 (DOCK8) in the autosomal-recessive form of hyper-IgE syndrome. AB - BACKGROUND: The genetic etiologies of the hyper-IgE syndromes are diverse. Approximately 60% to 70% of patients with hyper-IgE syndrome have dominant mutations in STAT3, and a single patient was reported to have a homozygous TYK2 mutation. In the remaining patients with hyper-IgE syndrome, the genetic etiology has not yet been identified. OBJECTIVES: We aimed to identify a gene that is mutated or deleted in autosomal recessive hyper-IgE syndrome. METHODS: We performed genome-wide single nucleotide polymorphism analysis for 9 patients with autosomal-recessive hyper-IgE syndrome to locate copy number variations and homozygous haplotypes. Homozygosity mapping was performed with 12 patients from 7 additional families. The candidate gene was analyzed by genomic and cDNA sequencing to identify causative alleles in a total of 27 patients with autosomal recessive hyper-IgE syndrome. RESULTS: Subtelomeric biallelic microdeletions were identified in 5 patients at the terminus of chromosome 9p. In all 5 patients, the deleted interval involved dedicator of cytokinesis 8 (DOCK8), encoding a protein implicated in the regulation of the actin cytoskeleton. Sequencing of patients without large deletions revealed 16 patients from 9 unrelated families with distinct homozygous mutations in DOCK8 causing premature termination, frameshift, splice site disruption, and single exon deletions and microdeletions. DOCK8 deficiency was associated with impaired activation of CD4+ and CD8+T cells. CONCLUSION: Autosomal-recessive mutations in DOCK8 are responsible for many, although not all, cases of autosomal-recessive hyper-IgE syndrome. DOCK8 disruption is associated with a phenotype of severe cellular immunodeficiency characterized by susceptibility to viral infections, atopic eczema, defective T cell activation and T(h)17 cell differentiation, and impaired eosinophil homeostasis and dysregulation of IgE. PMID- 20004783 TI - Global variations in prevalence of eczema symptoms in children from ISAAC Phase Three. AB - BACKGROUND: In 1999, The International Study of Asthma and Allergies in Childhood (ISAAC) Phase One reported the prevalence of eczema symptoms in 715,033 children from 154 centers in 56 countries by using standardized epidemiologic tools. OBJECTIVE: To update the world map of eczema prevalence after 5 to 10 years (ISAAC Phase Three) and include additional data from over 100 new centers. METHODS: Cross-sectional surveys using the ISAAC questionnaire on eczema symptoms were completed by adolescents 13 to 14 years old and by parents of children 6 to 7 years old. Current eczema was defined as an itchy flexural rash in the past 12 months and was considered severe eczema if associated with 1 or more nights per week of sleep disturbance. RESULTS: For the age group 6 to 7 years, data on 385,853 participants from 143 centers in 60 countries showed that the prevalence of current eczema ranged from 0.9% in India to 22.5% in Ecuador, with new data showing high values in Asia and Latin America. For the age group 13 to 14 years, data on 663,256 participants from 230 centers in 96 countries showed prevalence values ranging from 0.2% in China to 24.6% in Columbia with the highest values in Africa and Latin America. Current eczema was lower for boys than girls (odds ratio, 0.94 and 0.72 at ages 6 to 7 years and 13 to 14 years, respectively). CONCLUSION: ISAAC Phase Three provides comprehensive global data on the prevalence of eczema symptoms that is essential for public health planning. New data reveal that eczema is a disease of developing as well as developed countries. PMID- 20004786 TI - ORAI1 deficiency and lack of store-operated Ca2+ entry cause immunodeficiency, myopathy, and ectodermal dysplasia. AB - BACKGROUND: Defects in the development or activation of T cells result in immunodeficiency associated with severe infections early in life. T-cell activation requires Ca2+ influx through Ca2+-release activated Ca2+ (CRAC) channels encoded by the gene ORAI1. OBJECTIVE: Investigation of the genetic causes and the clinical phenotype of immunodeficiency in patients with impaired Ca2+ influx and CRAC channel function. METHODS: DNA sequence analysis for mutations in the genes ORAI1, ORAI2, ORAI3, and stromal interaction molecule (STIM) 1 and 2, as well as mRNA and protein expression analysis of ORAI1 in immunodeficient patients. Immunohistochemical analysis of ORAI1 tissue distribution in healthy human donors. RESULTS: We identified mutations in ORAI1 in patients from 2 unrelated families. One patient is homozygous for a frameshift nonsense mutation in ORAI1 (ORAI1-A88SfsX25), and a second patient is compound heterozygous for 2 missense mutations in ORAI1 (ORAI1-A103E/L194P). All 3 mutations abolish ORAI1 expression and impair Ca2+ influx and CRAC channel function. The clinical syndrome associated with ORAI1 deficiency is characterized by immunodeficiency with a defect in the function but not in the development of lymphocytes, congenital myopathy, and anhydrotic ectodermal dysplasia with a defect in dental enamel calcification. In contrast with the limited clinical phenotype, we found ORAI1 protein expression in a wide variety of cell types and organs. CONCLUSION: Ca2+ influx through ORAI1 is crucial for lymphocyte function in vivo. Despite almost ubiquitous ORAI1 expression, the channel has a nonredundant role in only a few cell types judging from the limited clinical phenotype in ORAI1-deficient patients. PMID- 20004787 TI - Eosinophilic gastrointestinal disease and peanut allergy are alternatively associated with IL-5+ and IL-5(-) T(H)2 responses. AB - BACKGROUND: Both anaphylactic food allergy and eosinophil-associated gastrointestinal disorders are associated with T(H)2 responses and food-specific IgE, yet they have very different clinical presentations. OBJECTIVE: To determine whether the clinical differences between anaphylactic food allergy and eosinophil associated gastrointestinal disorders are reflected in different T(H)2 responses to foods. METHODS: Subjects with peanut allergy (PA), subjects with allergic eosinophilic gastroenteritis (AEG), and nonatopic subjects were enrolled. Antigen specific IL-4, IL-5, IFN-gamma, and TNF T-cell responses to peanut, soy, and shrimp were measured by using intracellular cytokine staining and polychromatic flow cytometry. RESULTS: Two distinct subpopulations of T(H)2 cells were found: IL-5+ T(H)2 (IL-4+, IL-5+) and IL-5(-) T(H)2 (IL-4+, IL-5(-)) cells. Peanut specific IL-5+ T(H)2 cells were present at a 20-fold greater frequency in AEG versus PA (81 vs 4 per 10(6) CD4 cells; P = .05), whereas there were similar frequencies of IL-5(-) T(H)2 cells (67 vs 41 per 10(6)). For all foods, IL-5+ T(H)2 cells accounted for a significantly greater fraction of the antigen specific cells in AEG relative to PA (29% vs 4%; P < .0001). In PA but not AEG, IL-5(-) T(H)2 responses to peanut were highly correlated with peanut-specific IgE (r = 0.87 vs 0.55, respectively). All subject groups elicited similar very low magnitude T(H)1 responses to food antigens. CONCLUSION: T(H)2 responses are composed of 2 subpopulations: IL-5+ T(H)2 and IL-5(-) T(H)2 cells. IL-5+ T(H)2 food allergen-specific T cells are singularly associated with AEG, whereas PA is associated with a dominant IL-5(-) T(H)2 response. These results suggest heterogeneity within the T(H)2 cytokine response, with different T(H)2 responses alternatively favoring IgE-mediated or eosinophil-dominant immunopathology. PMID- 20004788 TI - Placental transfer of allergen-specific IgG but not IgE from a specific immunotherapy-treated mother. PMID- 20004789 TI - Diagnosis of primary immunodeficiency: let your eyes do the talking. PMID- 20004790 TI - Optimizing epigenetic therapy for myelodysplastic syndromes: issues and strategies. PMID- 20004791 TI - Decitabine treatment of patients with higher-risk myelodysplastic syndromes. AB - Since receiving US regulatory approval in 2006 for the treatment of patients with myelodysplastic syndromes, decitabine has also gained marketing approval in several other countries. Rates of complete remission (CR) reported for decitabine range from 9% in the US registration trial to 34% in a study of novel treatment schedules, while reported overall improvement rates range from 30% to 73%. Further studies are needed to determine the relative merits of decitabine compared with azacitidine, the optimal decitabine dose and schedule, how best to proceed following CR, molecular predictors of treatment response, and the possible role of this agent in combination regimens. PMID- 20004792 TI - Hypomethylating agents in myelodysplastic syndromes changing the inevitable: the value of azacitidine as maintenance therapy, effects on transfusion and combination with other agents. AB - The implication of DNA hypermethylation in the pathogenesis of myelodysplastic syndromes (MDS) provides a rationale for using hypomethylating agents such as azacitidine. Growing evidence suggests that azacitidine may reverse epigenetic gene silencing at specific genomic targets. AZA-001 established azacitidine as the first agent to provide a significant overall survival benefit in MDS patients. These data confirmed that azacitidine has a progressively cumulative effect on the MDS clone and support the value of maintenance therapy. Prolonged survival was independent of achieving complete response. Azacitidine in combination with histone deacetylase inhibitors might offer better efficacy by modulating the methylation and acetylation states of silenced genes. PMID- 20004793 TI - In vitro basis for treatment with hypomethylating agents and histone deacetylase inhibitors: can epigenetic changes be used to monitor treatment? AB - Hematopoietic disorders such as myelodysplastic syndromes (MDS) show a high frequency of methylation of tumor suppressor genes. DNA methyltransferase (DNMT) inhibitors such as azacitidine and decitabine are used to target DNA methylation in MDS patients. Combining these drugs with histone deacetylase (HDAC) inhibitors in vitro resulted in synergistic tumor suppressor gene re-expression. Several phase I trials have examined methylation, gene expression and DNA damage as markers of clinical response to DNMT and HDAC inhibitors, with conflicting results. Trials are ongoing to investigate early methylation changes and DNA damage markers to understand the mechanisms of these drugs and as potential predictors of clinical response. PMID- 20004794 TI - Azacitidine in lower-risk myelodysplastic syndromes. AB - Azacitidine is the first agent to significantly prolong overall survival (OS) compared with conventional care regimens in patients with higher-risk myelodysplastic syndromes (MDS). Here, we review currently available data on azacitidine treatment in lower-risk MDS. In a phase III study, a subset of patients with lower-risk MDS treated with azacitidine achieved an overall response rate (ORR) of 60% and a longer median OS compared with supportive care (44 vs 27 months). In a phase II study investigating various azacitidine dose schedules, the hematological improvement rate was approximately 50% in lower-risk patients; most patients who achieved transfusion independence were also lower risk. A further retrospective study reported an ORR of 52% in MDS lower-risk patients who received > or = 4 cycles of therapy. Preliminary data for azacitidine combination therapy with erythropoietin, granulocyte colony stimulating factor, and valproic acid are intriguing but should be treated with caution. These results suggest the feasibility and effectiveness of azacitidine in the treatment of patients with lower-risk MDS. PMID- 20004795 TI - Epigenetic therapy for myelodysplastic syndromes has entered center stage. PMID- 20004796 TI - Review of azacitidine trials in Intermediate-2-and High-risk myelodysplastic syndromes. AB - The efficacy of azacitidine, a cytosine nucleoside analog, has been demonstrated in two large phase III trials in myelodysplastic syndromes (MDS) patients, primarily in higher-risk patients. In the Cancer and Leukemia Group B (CALGB) study (lower-and higher-risk MDS patients; n = 191) azacitidine significantly prolonged median time-to-progression to acute myeloid leukemia or death (21 vs 13 months for supportive care; p = 0.007). The AZA-001 trial (Intermediate-2-and High-risk MDS patients; n = 358) showed azacitidine significantly prolonged median overall survival compared with conventional care regimens (24.5 vs 15.0 months; p = 0.0001). These results establish azacitidine as a reference first line treatment in patients with Intermediate-2-and High-risk MDS who are not immediate candidates for allogeneic stem cell transplantation. PMID- 20004797 TI - Health care transitions: a review of integrated, integrative, and integration concepts. AB - In this article, several views of the terms integration, integrated, and integrative are considered with the hopes that this brief review will help to raise awareness, clarify various uses of these terms, and add to the continuing discussion of integration and how we might improve health care. Models of integrative care, views of integration, and samples of different interpretations and definitions are offered. PMID- 20004798 TI - The difference between integration and collaboration in patient care: results from key informant interviews working in multiprofessional health care teams. AB - OBJECTIVES: Despite the growing interest in integrative health care, collaborative care, and interdisciplinary health care teams, there appears to be little consistency in terminology and clarity regarding the goal for these teams, other than "working together" for the good of the patients. The purpose of this study was to explore what the terms integration and collaboration mean for practitioners and other key informants working in multiprofessional health care teams, with a specific look at chiropractic and family physician teams in primary care settings. METHODS: Semistructured interviews were conducted with 16 key informants until saturation was obtained in the key emerging themes. All interviews were audiorecorded, and the transcripts were coded using qualitative content analysis. RESULTS: Most participants differentiated collaboration from integration. They generally described a model of professions working closely together (ie, collaborating) in the delivery of care but not subsumed into a single organizational framework (ie, integration). Our results suggest that integration requires collaboration as a precondition but collaboration does not require integration. CONCLUSIONS: Collaboration and integration should not be used interchangeably. A critical starting point for any new interdisciplinary team is to articulate the goals of the model of care. PMID- 20004799 TI - A nonsurgical approach to the management of patients with lumbar radiculopathy secondary to herniated disk: a prospective observational cohort study with follow up. AB - OBJECTIVE: This study presents the outcomes of patients with lumbar radiculopathy secondary to disk herniation treated after a diagnosis-based clinical decision rule. METHODS: A prospective observational cohort study was conducted at a multidisciplinary, integrated clinic that includes chiropractic and physical therapy health care services. Data on 49 consecutive patients were collected at baseline, at the end of conservative, nonsurgical treatment and a mean of 14.5 months after cessation of treatment. Disability was measured using the Bournemouth Disability Questionnaire (BDQ) and pain using the Numerical Rating Scale for pain. Fear beliefs were measured with the Fear-Avoidance Beliefs Questionnaire (FABQ). Patients also self-rated improvement. RESULTS: Mean duration of complaint was 60.5 weeks. Mean self-rated improvement at the end of treatment was 77.5%. Improvement was described as "good" or "excellent" in nearly 90% of patients. Mean percentage improvement on the BDQ was 60.4%. Numerical Rating Scale improved 4.1 points and FABQ improved 4.8 points. Clinically meaningful improvements in pain and disability were seen in 79% and 70% of patients, respectively. Mean number of visits was 13.2. After an average long term follow-up of 14.5 months, mean self-rated improvement was 81.1%. "Good" or "excellent" improvement was reported by 80% of patients. Mean percentage improvement in BDQ was 67.4%. Numerical Rating Scale improved 4.2 points and FABQ 4.5 points. Clinically meaningful improvements in pain and disability were seen in 79% and 73% of patients, respectively. CONCLUSIONS: Management based on the decision rule yielded favorable outcomes in this cohort study. Improvement appeared to be maintained over the long term. PMID- 20004800 TI - Cost minimization analysis of low back pain claims data for chiropractic vs medicine in a managed care organization. AB - OBJECTIVE: A managed care organization (MCO) examined differences in allowed cost for managing low back pain by medical providers vs chiropractors in an integrated care environment. The purpose of this study is to provide a retrospective cost analysis of administrative data of chiropractic vs medical management of low back pain in a managed care setting. METHODS: All patients with a low back pain related diagnosis presenting for health care from January 2004 to June 2004 who were insured by an MCO in northeast Wisconsin were tracked. The cumulative health care costs incurred by this MCO during the 2-year period from January 2004 to December 2005 related to these back pain diagnoses were collected. RESULTS: Allowed costs of chiropractic treatment were 12% greater than medical primary care and 60% less per case than other types of medical care combined, on a per case basis: median cost of medical primary care was $365.00, chiropractic care was $417.00, and medical nonprimary care was $669.00. CONCLUSION: This study of an MCO's low back pain allowed costs may be better redirected to primary care or chiropractic, given equivalent levels of case complexity. This study suggests chiropractic management as less expensive compared with medical management of back pain when care extends beyond primary care. Primary care management alone is virtually indistinguishable from chiropractic management in terms of costs. PMID- 20004801 TI - Hospital-based chiropractic integration within a large private hospital system in Minnesota: a 10-year example. AB - OBJECTIVE: The purpose of this article is to describe a model of chiropractic integration developed over a 10-year period within a private hospital system in Minnesota. METHODS: Needs were assessed by surveying attitudes and behaviors related to chiropractic and complementary and alternative medicine (CAM) of physicians associated with the hospital. Analyzing referral and utilization patterns assessed chiropractic integration into the hospital system. RESULTS: One hundred five surveys were returned after 2 mailings for a response rate of 74%. Seventy-four percent of respondents supported integration of CAM into the hospital system, although 45% supported the primary care physician as the gatekeeper for CAM use. From 2006 to 2008, there were 8294 unique new patients in the chiropractic program. Primary care providers (medical doctors and physician assistants) were the most common referral source, followed by self-referred patients, sports medicine physicians, and orthopedic physicians. Overall examination of the program identified that facilitators of chiropractic integration were (1) growth in interest in CAM, (2) establishing relationships with key administrators and providers, (3) use of evidence-based practice, (4) adequate physical space, and (5) creation of an integrated spine care program. Barriers were (1) lack of understanding of chiropractic professional identity by certain providers and (2) certain financial aspects of third-party payment for chiropractic. CONCLUSION: This article describes the process of integrating chiropractic into one of the largest private hospital systems in Minnesota from a business and professional perspective and the results achieved once chiropractic was integrated into the system. This study identified key factors that facilitated integration of services and demonstrates that chiropractic care can be successfully integrated within a hospital system. PMID- 20004802 TI - An analysis of the integration of chiropractic services within the United States military and veterans' health care systems. AB - OBJECTIVES: The purpose of this article is to compare chiropractic integration within the health care systems of the Department of Defense and Department of Veterans Affairs and to identify practices and policies that may either support or challenge the extent of chiropractic integration within those systems. METHODS: As subject matter experts and providers within these systems, our team reviewed enacted legislation, policies, and the literature pertinent to chiropractic practice in Department of Defense and Department of Veterans Affairs medical facilities, and identified opportunities and threats pertinent to integration. RESULTS: We identified 9 areas wherein potential opportunities and threats to integration existed, including legislative history, programmatic growth, leadership structure, employment status of providers, clinical work duties, patient access, patient demographics, academic affiliations, and research. CONCLUSION: These findings provide a higher level of understanding regarding the current state and future direction of chiropractic service integration within these integrated health care systems. PMID- 20004803 TI - Measuring the development of integrative care communication patterns and cross disciplinary knowledge at a university clinic: a baseline cross-sectional study. AB - OBJECTIVE: The initiation of multiple clinical programs within a previously single-purpose health care institution brings multiple health care providers together under one roof. A baseline cross-sectional survey was undertaken to measure the development of integrative care communication patterns and cross disciplinary knowledge at a health sciences university clinic. METHODS: This study was conducted by means of 3 separate surveys administered to all complementary and alternative medicine (CAM) faculty clinicians, CAM student interns, and patients attending CAM therapy at a university clinic (National University of Health Sciences, Lombard, Ill). The CAM therapies included acupuncture and Oriental medicine, chiropractic medicine/doctor of chiropractic, massage therapy, and naturopathic medicine/naturopathic doctor. RESULTS: Consultations between practitioners, including clinicians and interns, of different professions were predominantly informal in nature. The highest level of familiarity was with chiropractic philosophy; the lowest level was with naturopathic philosophy. Clinicians were very familiar with the philosophies of the other professions, with interns and patients being variably familiar with the other professions' philosophies. Less than 20% of patients reported being referred to another care provider. Clinicians, interns, and patients indicated that they would like additional opportunities to take advantage of multiple professions working together. CONCLUSIONS: This study is an initial assessment of the knowledge, communication patterns, and actions within the clinic during these early stages of integration. Recommendations for integrating both clinical care and educational processes within a multiple professions university are considered. PMID- 20004804 TI - Functional scores and subjective responses of injured workers with back or neck pain treated with chiropractic care in an integrative program: a retrospective analysis of 100 cases. AB - OBJECTIVE: The purpose of this study is to report on integrative care for the treatment of injured workers with neck or back pain referred to a doctor of chiropractic from a medical or osteopathic provider. METHODS: This retrospective case series study evaluated data on 100 patients referred for chiropractic care of work-related spinal injuries involving workers' compensation claims. Deidentified data included age, sex, visual analog scale scores for pain, pre- and posttreatment Functional Rating Index (FRI) scores, and subjective response to chiropractic care. Based on date of injury to first chiropractic treatment, patients were subdivided as acute, subacute, or chronic injured workers. Cases were analyzed for differences in pretreatment FRI scores, posttreatment FRI scores, FRI change scores (posttreatment FRI minus pretreatment FRI score), and subjective percentage improvement using a 1-way analysis of variance. Treatment included manual therapy techniques and exercise. RESULTS: Injured workers with either an acute or subacute injury had significantly lower posttreatment FRI scores compared with individuals with a chronic injury. The FRI change scores were significantly greater in the acute group compared with either the subacute or chronic injured workers. Workers in all categories showed improved posttreatment tolerance for work-related activities and significantly lower posttreatment subjective pain scores. CONCLUSIONS: The study identified positive effects of chiropractic management included in integrative care when treating work-related neck or back pain. Improvement in both functional scores and subjective response was noted in all 3 time-based phases of patient status (acute, subacute, and chronic). PMID- 20004805 TI - Demographic characteristics of 38 patients injured in motor vehicle accidents referred by chiropractors to physiatrists. AB - OBJECTIVE: The purpose of this study is to describe the demographic profile of patients in the New Jersey area who are involved in motor vehicle personal injury lawsuits and who are referred from chiropractors to physiatrists. METHODS: The study design was a prospective chart review of patients (N = 38) referred to a private physiatric practice from 5 chiropractic practices. Patient data collected at initial consultation included age, employment status, emergency department consultation, time since accident, visual analog score, neck pain and back pain, review of systems, and functional limitations. RESULTS: The average patient was 37.1 years old, with male-to-female ratio nearly 1:1, and presenting 4.5 months after the accident; 81.6% were employed before the accident, 25.8% of which stopped working. The average pain score was 6.6 on a visual analog scale. Neck and back pain were common at 84.2% and 89.5%, respectively. Other complaints included headaches, sleeping difficulties, dizziness, depression, and anxiety. Limitations in function was reported in most patients. CONCLUSIONS: In this study, patients referred to a physiatrist from doctors of chiropractic had neck and low back pain not requiring hospital admission. Patients referred tended to have complicated cases with a variety of medical, legal, and psychological factors that are associated with delayed recovery. Physiatrists may be uniquely suited to assist chiropractors in management of complicated patients who have been involved in motor vehicle personal injury lawsuits and who have multidisciplinary needs. PMID- 20004806 TI - A case of cerebellopontine angle meningioma presenting with neck and upper extremity pain. AB - OBJECTIVE: The purpose of this case report is to describe and discuss the clinical presentation, diagnosis, and management of a patient with a cerebellopontine angle meningioma. CLINICAL FEATURES: A 29-year-old man presented to a chiropractor with diffuse musculoskeletal pain in the neck and right upper extremity. The findings of the clinical examination included a unilateral lower cranial nerve motor deficit. INTERVENTION AND OUTCOME: Magnetic resonance imaging studies of the neck and brain revealed a posterior fossa tumor, which was eventually diagnosed as a benign meningioma. Partial surgical removal of the tumor mass was followed by radiation therapy. Postoperative morbidity was fairly low. Lower cranial nerve function normalized after surgery. CONCLUSION: Patients with brain lesions may present to chiropractic practices with predominantly musculoskeletal symptoms. Chiropractors can aid in the multidisciplinary and integrated management of such conditions through careful interviewing and appropriate neurological examination. PMID- 20004807 TI - Integrated musculoskeletal rehabilitation care at a comprehensive combat and complex casualty care program. AB - OBJECTIVE: The purpose of this study is to describe the musculoskeletal rehabilitation model used to care for combat and severely wounded or ill US military service members at an integrated Comprehensive Combat and Complex Casualty Care center located at Naval Medical Center San Diego. METHODS: Through a collaborative and iterative process, providers from the various services included at the Comprehensive Combat and Complex Casualty Care program developed a description of the integration of services provided at this location. RESULTS: After construction of the facility in 2007, the program has provided services for approximately 2 years. Eighteen different health care providers from 10 different specialties provide integrated musculoskeletal services, which include primary care, physical therapy, occupational therapy, vestibular therapy, gait analysis, prosthetics, recreational therapy, and chiropractic care. At the time of this writing (early 2009), the program had provided musculoskeletal rehabilitation care to approximately 500 patients, 58 with amputations, from the operational theater, Veterans Affairs, other military treatment facilities, and local trauma centers. CONCLUSION: The complex nature of combat wounded and polytrauma patients requires an integrated and interdisciplinary team that is innovative, adaptable, and focused on the needs of the patient. This article presents a description of the model and the experiences of our musculoskeletal rehabilitation team; it is our hope that this article will assist other centers and add to the small but emerging literature on this topic. PMID- 20004808 TI - Contextualizing integration: a critical social science approach to integrative health care. AB - This article argues for the importance of examining the phenomenon of integrative health care in broader social and historical contexts. The authors examine mainstream approaches to identify patterns of integrative medicine and criticize them for their neglect of clashes among different philosophical paradigms and the wider social contexts that govern health care in practice. The authors outline a framework and highlight the values of a critical social science perspective in deepening our understanding of recent transformations in health care practice and issues surrounding biomedicine and complementary/alternative medicine (including chiropractic, naturopathy, massage, acupuncture/oriental medicine, etc) and traditional medicine. A critical social science perspective pays special attention to complex power relations, inclusionary/exclusionary strategies, and interprofessional dynamics in medicine. Drawing upon recent research findings, the authors illustrate how such a perspective reveals the intricacies and tensions that surround the integration of different paradigms of health care practice. The authors summarize the importance of situating integrative health care in structural contexts and affirm their commitment to a critical social science approach. PMID- 20004809 TI - Chiropractic practice in the Danish public health care sector: a reflection and clarification of context, terms of usage, and selected design considerations for a planned qualitative investigation. AB - This commentary offers preliminary considerations around a phenomenological investigation of chiropractic services in a Danish public sector setting. In this narrative description, the main venue for chiropractic public (secondary) sector practice in the Danish context is briefly described and defined. Furthermore, a contextually relevant definition of an integral health care service is presented; and the professional importance for chiropractic in providing such services is also discussed. Finally, salient questions requiring empirical investigation in this context are posed; and selected issues around a qualitative research design approach are considered. PMID- 20004810 TI - Infection preventionists' experience during the first months of the 2009 novel H1N1 influenza A pandemic. AB - BACKGROUND: A novel strain of influenza A (H1N1) was identified in April 2009 and developed into a pandemic by June 2009. This rapid and unexpected event had enormous implications for infection preventionists (IP) internationally. Lessons learned from this event should guide future pandemic planning efforts. METHODS: Focus groups were conducted at the Association for Professionals in Infection Control and Epidemiology, Inc, (APIC) 2009 conference to evaluate IPs' experience with the novel H1N1 influenza pandemic and assess their perceived needs related to novel H1N1 topics and products required for future education and reference materials. RESULTS: Forty IPs (37 from the United States and 3 international) participated in the focus groups. Needed reference materials identified by attendees included infection prevention guidance for nonacute care settings; occupational health polices; and brief, multilanguage patient/family educational materials. Educational topics on which IPs need to be trained include isolation precautions/personal protective equipment recommendations for novel H1N1 patients, coordination between hospitals and community response agencies, and surge management. The rapidly changing and conflicting recommendations related to patient management made responding to this event challenging. IPs require synthesized infection prevention guidelines developed in a concise, real-time format. CONCLUSION: IPs must continue to partner with public health and other response agencies to address gaps in pandemic planning. PMID- 20004811 TI - National Healthcare Safety Network (NHSN) report: data summary for 2006 through 2008, issued December 2009. PMID- 20004812 TI - The World Health Organization hand hygiene observation method. AB - Monitoring hand hygiene adherence and providing performance feedback to health care workers is a critical component of multimodal hand hygiene promotion programs, but important variations exist in the way adherence is measured. Within the framework of the World Health Organization's (WHO) First Global Patient Safety Challenge known as "Clean Care is Safer Care," an evidence-based, user centered concept, "My five moments for hand hygiene," has been developed for measuring, teaching, and reporting hand hygiene adherence. This concept is an integral part of the WHO's hand hygiene improvement strategy conceived to translate the WHO Guidelines on Hand Hygiene in Health Care into practice. It has been tested in numerous health care facilities worldwide to ensure its applicability and adaptability to all settings irrespective of the resources available. Here we describe the WHO hand hygiene observation method in detail-the concept, the profile and the task of the observers, their training and validation, the data collection form, the scope, the selection of the observed staff, and the observation sessions-with the objective of making it accessible for universal use. Sample size estimates, survey analysis and report, and major bias and confounding factors associated with observation are discussed. PMID- 20004813 TI - The impact of a screensaver in promoting hand hygiene. PMID- 20004814 TI - Medical students need more education on hygiene behavior. PMID- 20004815 TI - Swine flu infection among medical students: an issue of concern. PMID- 20004816 TI - Can influenza A be transmitted from patients previously vaccinated against influenza? PMID- 20004819 TI - On the tech horizon: new software leads to decreased patient mortality. PMID- 20004820 TI - Can't stand the suspense. PMID- 20004821 TI - Can't stand the suspense. PMID- 20004822 TI - Can't stand the suspense. PMID- 20004823 TI - Takes all kinds. PMID- 20004824 TI - Media 'trauma': new NBC series gives managers something to think about. PMID- 20004825 TI - Pearls: stuff you gotta know. PMID- 20004826 TI - Hidden trauma: seizure patients have a significant risk for injury. PMID- 20004827 TI - Does size matter? Paramedic program class size may predict success. PMID- 20004828 TI - Stretch your routine: athletes aren't the only ones to benefit from flexibility. PMID- 20004829 TI - Back to basics: the lost art of BLS care. PMID- 20004830 TI - Toward the sound of shooting: Arlington county, VA., rescue task force represents a new medical response model to active shooter incidents. PMID- 20004831 TI - Instant feedback: point-of-care testing helps providers make life-saving decisions. PMID- 20004832 TI - Faith in a cure: understanding the role folk medicine plays in prehospital care of many Mexican-Americans. PMID- 20004833 TI - Dag nab it! Why didn't I think of that? PMID- 20004834 TI - [Detection and quantification of apoptosis in tissues: a challenge]. PMID- 20004835 TI - [Detection of apoptosis in vivo: comparison of different methods in histological sections of subcutaneous xenografts of HT29 human colon adenocarcinoma]. AB - Apoptosis detection in histological section is very important, but usual methods (TUNEL and morphologic changes) are controversial. Immunohistochemical stains of activated proteins during apoptosis improve detection of cell death in tissue sections. Activated caspase-3 and cleaved cytokeratin-18 are more and more used. OBJECTIVE: This study compared immunohistochemical markers (activated caspase-3, cleaved cytokeratin-18 and two antibodies not yet evaluated: activated caspase-7 and cleaved PARP-1), cellular morphology and TUNEL. MATERIAL AND METHODS: Tumour xenografts of the human colon cancer cell line HT29 were used, treated by photodynamic therapy, to obtain large numbers of cells undergoing apoptosis. Apoptotic cells were quantified and apoptotic indices were determined for each marker. RESULTS: Comparison of apoptosis index indicated statistically best sensibility with activated-caspase-3 and cleaved-cytokeratin-19. CONCLUSION: Immunohistochemistry for activated caspase-3 and cleaved cytkeratin-18 appear to be an easy, sensitive, and reliable method for detecting and quantifying apoptosis in this model. There are therefore recommended for the detection and quantification of apoptosis in tissue sections. Other markers as cleaved PARP-1 and activated caspase-7 can be an interessant solution: advantages and inconvenience for each methods are exposed. PMID- 20004837 TI - [Conjunctiva tumors of the skin: "I look and I do not know!"]. PMID- 20004836 TI - [Lymphoid hemopathies and HHV-8]. AB - HHV-8 is a gamma herpes virus described for the first time in humans in 1994 within Kaposi's disease cells. Since, it has been found to be involved in lymphoid proliferative disorders. First, it has been described in multicentric Castleman's disease (100 % of HIV infected patients and 50 % of HIV negative patients), in multicentric Castleman's disease associated lymphomas and in cavitary and extracavitary primary effusion lymphomas (100 % of cases). More recently, some other HHV-8 associated lymphoproliferations have been described. We describe in this review the characteristics of this virus, along with the clinical, pathological and molecular characteristics of the different entities associated with HHV-8. PMID- 20004838 TI - [Connective tissue tumors of the skin. Plexiform fibro-histiocytic tumor]. PMID- 20004839 TI - [Connective tissue tumors of the skin. Giant cell fibroblastoma]. PMID- 20004840 TI - [Mesenchymal tumors of the skin. Cellular neurothecoma]. PMID- 20004841 TI - [Mesenchymal tumors of the skin. Atypical vascular proliferation after radiotherapy]. PMID- 20004842 TI - [Mesenchymal tumors of the skin. Desmoplastic leiomyosarcoma]. PMID- 20004843 TI - [Mesenchymal tumors of the skin. Acral superficial fibromyxoma]. PMID- 20004844 TI - [Mesenchymal tumors of the skin. Atypical fibrous histiocytoma]. PMID- 20004845 TI - [Mesenchymal tumors of the skin. Tufted angioma (angioblastoma)]. PMID- 20004846 TI - [Conclusion. Mesenchymal tumors of the skin: "what are the differences between cutaneous mesenchymal tumors and their counterparts of deep soft tissues?"]. PMID- 20004847 TI - [Lipomatous polyposis of the colon: a case report]. AB - Lipomatous polyposis of the colon is a rare affection, with about 10 published cases. This affection is characterized by a great number of polyps which can reach several hundreds, of diffuse location on the entire length of the colon, with a higher density on the left colon and the sigmoid. The size of polyps varies from 1 to 10 cm. When they are voluminous, they can be cause of obstruction, ulceration or bleeding. We report a case of colonic lipomatous polyposis, associated with an ileocolic lipomatous, an ileocolic diverticulosis and adenomatous polyps in a 70-year-old man. PMID- 20004848 TI - [Maze-like vascular anomaly in partial mole. Interest for the pathological diagnosis of partial mole on chorionic villous sampling]. AB - A case of maze-like angiomatoid anomaly in villi obtained by chorionic villous sampling (CVS) is described. This feature is pathognomonic of partial mole (triploid syndrome) and it was later confirmed by chromosomal analysis. Maze-like angiomatoid anomaly was previously described on specimen submitted after spontaneous or induced abortions, but it was never reported on CVS. This report emphasized that microscopic investigation of CVS cannot be conclusive for cytogenetic anomaly in almost all cases excepted for partial mole where diagnosis criteria are usually characteristic. PMID- 20004849 TI - [Myocardial involvement in invasive aspergillosis]. AB - We report the case of a 48-year-old female patient who had a Crohn's disease treated by corticosteroids. The patient developed severe cardiac failure, which was refractory to treatment with inotropic agents. At necropsy, examination of the heart revealed myocardial abscesses. On microscopic study, we diagnosed an aspergillar myocarditis. Aspergillar myocarditis is a rare and fatal localisation in disseminated aspergillosis. Diagnosis is difficult and treatment, usually initiated late, is ineffective. PMID- 20004850 TI - [An uncommon duodenal tumor]. PMID- 20004851 TI - [An unusual cause of chronic dysphagia]. PMID- 20004852 TI - [An unusual mastocytosis...]. PMID- 20004853 TI - [A rare tumour of ovary]. PMID- 20004854 TI - [Unusual polyp of the small intestine]. PMID- 20004855 TI - [An unusual hepatic tumor]. PMID- 20004856 TI - Childhood periodic syndromes. AB - This review focuses on so-called "periodic syndromes of childhood that are precursors to migraine," as included in the second edition of the International Classification of Headache Disorders. Presentation is characterized by an episodic pattern and intervals of complete health. Benign paroxysmal torticollis is characterized by recurrent episodes of head tilt, secondary to cervical dystonia, with onset between ages 2-8 months. Benign paroxysmal vertigo presents as sudden attacks of vertigo lasting seconds to minutes, accompanied by an inability to stand without support, between ages 2-4 years. Cyclic vomiting syndrome is distinguished by its unique intensity of vomiting, affecting quality of life, whereas abdominal migraine presents as episodic abdominal pain occurring in the absence of headache. Their mean ages of onset are 5 and 7 years, respectively. Diagnostic criteria and appropriate evaluation represent the key issues. Therapeutic recommendations include reassurance, lifestyle changes, and prophylactic as well as acute antimigraine therapy. PMID- 20004857 TI - Epilepsy surgery outcomes: quality of life and seizure control. AB - A consecutive, retrospective analysis of seizure control and quality of life was performed among 83 pediatric patients undergoing epilepsy surgery at Children's Hospital of Wisconsin. Seizure outcomes were generally favorable, with 68.7% class I outcomes; class II, 12%; and class III, 19.3%. Seizure freedom was highest among temporal lobectomies (84.2%) and hemispherectomies (76.2%). Outcomes among hemispherectomies were substantially superior to those of multilobar resections. Cortical dysplasia was associated with lower seizure freedom, at 57.5%. Among age groups, seizure-free outcomes in infants were lowest, at 50%. The lower infant seizure-free rate was likely attributable to frequency of multilobar resections and type of pathology (cortical dysplasia). Quality-of-life measures generally paralleled seizure outcomes. These results indicate that epilepsy surgery in children with intractable epilepsy can result in significant improvements in seizure control, quality of life, and development. Anticipated type of surgery, presumed location of epileptogenic site, absence of a defined lesion on magnetic resonance imaging scan of the brain, and patient's age should not prevent surgical evaluations of children with intractable epilepsy. PMID- 20004858 TI - Adrenocorticotropic hormone versus pulsatile dexamethasone in the treatment of infantile epilepsy syndromes. AB - For treatment of intractable epilepsies, there are no data comparing conventional adrenocorticotropic hormone and pulsatile corticoid therapy with dexamethasone. A retrospective comparison of efficacy was therefore conducted for both forms of application. Between 1989 and 2001, a series of 11 children with West syndrome and 3 with Lennox-Gastaut syndrome were treated with adrenocorticotropic hormone (group 1); between 2003 and 2006, 7 children with West syndrome, 5 with electrical status epilepticus during slow sleep, and 2 with Lennox-Gastaut syndrome were treated with pulsatile corticoid therapy (group 2). In group 1 (n = 14), 9/11 West syndrome patients became seizure free, but none with Lennox Gastaut syndrome (0/3). In group 2 (n = 14), 4/7 West syndrome patients became seizure-free, 1/2 with Lennox-Gastaut syndrome exhibited seizure-frequency reduction, and 2/5 patients with electrical status epilepticus during slow-wave sleep exhibited significant improvement according to electroencephalograms. In West syndrome, pulsatile corticoid therapy was an effective alternative treatment to adrenocorticotropic hormone, whereas in Lennox-Gastaut syndrome in general steroids did not lead to a significant seizure reduction. In electrical status epilepticus during slow-wave sleep, treatment with pulsatile corticoid therapy seems to be effective and should be investigated in a larger group of patients. PMID- 20004859 TI - Febrile seizures and primary human herpesvirus 6 infection. AB - Primary human herpesvirus 6 infection is acquired mainly during the first two years of life and is often associated with febrile seizures. The aim of the present study was to investigate in Greece the frequency and clinical characteristics of primary human herpesvirus 6 (HHV-6) infection in hospitalized children with febrile seizures. Children aged from 6 months to 5 years without known neurologic disease were examined for primary HHV-6 infection, by real-time polymerase chain reaction in acute-phase plasma and by indirect immunofluorescent assay for antibody titers in acute and convalescent serum. Of 65 children included in the analysis, 55 experienced the first febrile episode of seizures and 10 the second. Primary HHV-6 infection was verified in 10 of 55 children with a first febrile episode (18%), whereas none of the 10 children with a second episode of seizures had primary HHV-6 infection. Eight children were infected with HHV-6 type B and two with type A. None of the 85 control subjects had primary HHV-6 infection, but 49% had immunoglobulin G antibodies against the virus. These findings suggest that primary HHV-6 infection is frequently associated with febrile seizures in children in this geographic region and should be considered, especially for a first episode of febrile seizures. PMID- 20004860 TI - Clinical and imaging findings suggesting human herpesvirus 6 encephalitis. AB - We sought to distinguish patients testing positive for human herpesvirus 6 from those testing negative, based on clinical features and magnetic resonance images. Sixteen immunosuppresed patients were tested by polymerase chain reaction for human herpes virus 6 DNA in cerebrospinal fluid (nine positive results). Medical records were examined for agitation, altered mental status, hallucinations, insomnia, memory loss, and seizures. Patients were sorted by viral status. Clinical features were compared with imaging findings. Insomnia, agitation, and hallucinations were preferentially evident in human herpes virus 6-positive patients. Imaging abnormalities were evident in the hippocampus of both groups. However, extrahippocampal involvement was more common in human herpes virus 6 positive patients and among those with insomnia and hallucinations or seizures. Patients with memory loss and imaging abnormalities in the entorhinal cortex or amygdala were likely to test positive, as were patients with hallucinations and abnormal magnetic resonance signal in the hippocampus. Human herpes virus 6 encephalitis patients present with diverse clinical features that are also common among patients who test negative. This entity should be suspected in patients who present with insomnia, seizures, or hallucinations when imaging abnormalities are evident in the hippocampus, amygdala, and limbic structures beyond the medial temporal lobe. PMID- 20004861 TI - Neurologic varicella complications before routine immunization in Germany. AB - Varicella is an acute febrile, highly infectious disease. We describe the incidence and types of neurologic complications in children up to 16 years old. Hospitalized varicella cases were prospectively captured by active nationwide surveillance through the German Pediatric Surveillance Unit for Rare Diseases from January 2003 to December 2004. Neurologic complications occurred in 232 (25.4%) of 918 hospitalized children with varicella, and were the most frequent reason for hospitalization. The median age was 4.2 years (interquartile range 2.5 5.9). The median duration of hospital stay was 6 days (interquartile range 3-11). Neurologic complications were more frequent (P=0.054) in immunocompetent (32%) than immunocompromised (4%) children. The most frequent diagnoses comprised acute cerebellar ataxia in 72 (31.0%), febrile convulsion in 69 (29.7%), meningoencephalitis in 52 (22.4%), cerebral convulsions in 21 (9.1%), syncope in 9 (3.9%), and cerebral vasculitis/infarction in 6 (2.6%) of all children with neurologic complications. Twenty-eight (12%) demonstrated sequelae (18 with ataxia, four with epilepsy, two with hemiparesis, three with cerebral nerve palsy, and one with dysesthesia). Three patients died. The yearly incidence of neurologic varicella-associated hospitalizations was estimated at 2.4 neurologic complications per 100,000 children, corresponding to about one neurologic complication in 2000 varicella cases. PMID- 20004862 TI - Diffusion tensor imaging in arginase deficiency reveals damage to corticospinal tracts. AB - Individuals with a proximal urea cycle disorder, such as carbamoyl phosphate synthetase deficiency 1 or ornithine transcarbamylase deficiency, may present with encephalopathy resulting from hyperammonemia. The clinical presentation of arginase deficiency is considerably different, characterized by progressive spasticity involving the lower extremities and usually dementia. Diagnosis may be delayed, and patients are often thought to have cerebral palsy. The true etiology of brain injury in arginase deficiency is unknown, but is not thought to be due to hyperammonemia and brain swelling, the mechanism of injury recognized in ornithine transcarbamylase deficiency. Elevated arginine could augment nitric oxide synthesis, leading to oxidative damage. The hypothesis for the present study was that specific brain vulnerability in arginase deficiency would involve microstructural alterations in corticospinal tracts and that this finding, as measured by diffusion tensor imaging, would differ from age-matched control subjects and those with ornithine transcarbamylase deficiency. Diffusion tensor imaging data were compared for a 17-year-old male patient with arginase deficiency, age-matched normal control subjects, and age-matched individuals with ornithine transcarbamylase deficiency. Significant differences were found in suspected areas of interest, specifically in the corticospinal tracts. This finding confirms the hypothesis that the mechanism of injury in arginase deficiency, although still unknown, is unlikely to be similar to that causing ornithine transcarbamylase deficiency. PMID- 20004864 TI - Idiopathic intracranial hypertension without papilledema. AB - Idiopathic intracranial hypertension is characterized by high cerebrospinal fluid pressure with no underlying structural or systemic cause. Idiopathic intracranial hypertension without papilledema, although well-described in adults, is rarely reported in the pediatric population. The usual presentation is similar to that of chronic daily headache, with some features of migraine. However, treatment modalities are different, and specific therapy can lead to significant improvement in symptoms. We describe six children with chronic daily headache, who were diagnosed with idiopathic intracranial hypertension without papilledema. The response to medical management was variable. One child required a lumboperitoneal shunt for persistent signs, with good surgical outcome. PMID- 20004863 TI - Barkhof magnetic resonance imaging criteria predict early relapse in pediatric multiple sclerosis. AB - We sought to identify clinical and radiologic features predicting early relapse after a diagnosis of multiple sclerosis in children. In this nationwide retrospective multicenter study in The Netherlands, we included 28 children with multiple sclerosis with onset before age 16 years. Magnetic resonance images and clinical features at the onset of disease were evaluated. The mean follow-up time was 55 months. Twenty children (71%) had a relapse during follow-up. We found that the presence of at least three of four Barkhof magnetic resonance imaging criteria at the onset of multiple sclerosis signs is predictive of early relapse after a diagnosis of multiple sclerosis in children (P<0.05). PMID- 20004865 TI - Fetal MRI of a hypothalamic hamartoma in Pallister-Hall syndrome. AB - Fetal magnetic resonance imaging is increasingly being used as an adjunct to ultrasound. It allows for better visualization of in utero brain development and intracranial abnormalities (especially cerebral malformations). Hypothalamic hamartoma is a nonneoplastic malformation resembling normal hypothalamic tissue both histologically and on magnetic resonance imaging. Although it is rare, this entity is important to recognize for appropriate management and genetic counseling. We describe a unique patient in whom magnetic resonance imaging of the fetal brain allowed a prenatal diagnosis of Pallister-Hall syndrome. PMID- 20004866 TI - Epileptic apnea in a trisomy 18 infant. AB - Trisomy 18 is frequently associated with neurologic abnormalities, including hypotonia in infancy, mental retardation, central apnea, and epilepsy. Although central apnea and epilepsy are common complications in children with trisomy 18, epileptic apnea in these children was not previously reported. We describe an infant with trisomy 18 who developed epileptic apnea. Her apneic episodes began at age 10 months, occurring in clusters over several days each month. According to ictal electroencephalogram, interictal [(11)C] flumazenil-positron emission tomography, and [(18)F] fluorodeoxyglucose-positron emission tomography, the apneic episodes represented complex partial seizures (autonomic seizures), probably originating in the left frontotemporal area, probably related to cortical microdysgenesis. The condition was successfully treated with zonisamide. In infants with trisomy 18, differentiation of epileptic apnea from central apnea is crucial, because medications used to treat central apnea, including caffeine and theophylline, can be harmful to children with epileptic apnea. PMID- 20004868 TI - Neonatal spinal cord injury after an uncomplicated vaginal delivery. AB - Neonatal spinal cord injury has been reported after traumatic births and as a consequence of underlying lesions in the spinal cord. This report describes an infant who was born with bilateral flaccid paralysis of the upper extremities after an atraumatic, noninstrumented vaginal delivery. The infant was otherwise neurologically intact. The infant was initially thought to exhibit bilateral brachial plexus injury. However, magnetic resonance imaging demonstrated an upper cervical spinal cord hemorrhage, with no underlying lesions of the spinal cord or surrounding vasculature. This case highlights the importance of thoroughly evaluating any neurologic deficit in the newborn, and suggests that normal mechanical forces of labor and delivery may be sufficient to cause damage to the newborn spinal cord. PMID- 20004867 TI - Gaucher disease patient with myoclonus epilepsy and a novel mutation. AB - The N188S mutation in Gaucher disease is associated with myoclonus epilepsy. We performed genetic analysis on a patient with progressive myoclonus epilepsy, who had received antiepileptic drugs for over 10 years. We detected N188S/G199D on the gene encoding glucocerebrosidase. Mutant proteins carrying each mutation were expressed in COS-1 cells (a commonly used cell line which derives from kidney cells of the African green monkey). Measurements of enzymatic activity and Western blotting analysis were performed. When residual activities were measured, glucocerebrosidase with the N188S mutation exhibited 50% activity of the wild type, and with G199D, 7.4%. Neither mutation influenced the stability of the enzyme protein. These data suggested a diagnosis of Gaucher disease for this patient, and indicated that G199D is a novel mutation. PMID- 20004869 TI - Giant, solid, invasive prolactinoma in a prepubescent boy with gynecomastia. AB - Pituitary adenomas are extremely rare in prepubescent children. Most of these tumors are functioning adenomas, but can also present with vision disturbances and changes in body morphology. We discuss the presentation, management, and outcome of a giant, solid, invasive prolactinoma in a 7-year-old boy who presented with visual disturbances and gynecomastia. The child demonstrated significant improvement in vision and a reduction in size of the tumor after subtotal tumor decompression and treatment with oral bromocriptine for 4 months. Tumor decompression and oral dopamine agonist therapy comprise the accepted treatment for pediatric prolactinomas. To our knowledge, only two previous reports described a giant prolactinoma in a prepubescent child. The relevant literature on pediatric pituitary tumours and prolactinoma is reviewed. PMID- 20004871 TI - Thwarted at every turn. PMID- 20004872 TI - The paraoptometric-optometric team. PMID- 20004873 TI - Alzheimer's disease: visual system review. AB - BACKGROUND: Ten million baby boomers in the United States will get Alzheimer's disease. Optometrists can benefit from understanding the impact the Alzheimer's disease process has on the visual system. This can result in more effective management of the condition and in more effective communication with members of the Alzheimer's disease multidisciplinary team. METHODS: This is a review of the literature but by no means a completely exhaustive review. Alzheimer's disease is a complex disease. A rapidly expanding body of knowledge covers multiple disciplines. RESULTS: The visual system shows deficits early in the degenerative process of Alzheimer's disease. Biomarkers through the visual system such as nerve fiber deficits, lens opacities, and functional losses in the magnocellular pathway, such as contrast sensitivity and temporal processing, may prove to not only help detect Alzheimer's disease early but also detect it before there are the classic cognitive and memory losses. CONCLUSIONS: The effects of Alzheimer's disease are devastating. Optometrists, as primary care clinicians, can make critical contributions in the diagnosis, treatment, and management of this neurodegenerative disease. PMID- 20004874 TI - Acute zonal occult outer retinopathy: a case report. AB - BACKGROUND: Acute zonal occult outer retinopathy (AZOOR) is a disorder of unknown etiology with a predilection for young women. AZOOR is characterized by an acute loss of one or more zones of outer retinal function with a corresponding loss of visual field in one or both eyes. Patients present with photopsia, variable funduscopic changes, and abnormal electroretinogram (ERG) findings. There are no proven treatments. Diagnosed cases have a reasonable prognosis because central vision is often spared. CASE REPORT: A 24-year-old white woman presented with acute photopsia with clouded temporal vision in the right eye (O.D.). Initial evaluation found a slightly swollen nerve fiber layer with no other outstanding lesions O.D. Over a 7-week follow-up period, there was significant progression into the deep chorioretinal tissues O.D. with an anterior chamber response. At 3 months, an afferent pupillary defect (APD) developed with significant retinal pigment epithelium (RPE) changes. After extensive testing, the consulting retinal specialist identified AZOOR as the etiology. At 21 and 31 months, the process remained in remission. Although primary symptoms subsided, the dense inferior temporal field defect remained O.D. CONCLUSION: AZOOR is a rare condition with subtle and often vague signs and symptoms making diagnosis difficult. The course of AZOOR, its clinical presentations, hypothesis of etiology, differential diagnosis, workup, management, and prognosis are reviewed. PMID- 20004875 TI - Correlation between asthenopic symptoms and different measurements of convergence and reading comprehension and saccadic fixation eye movements. AB - PURPOSE: The aim of this study was to evaluate the relationship between asthenopic symptoms, convergence amplitude, reading comprehension, and saccadic eye movements in children 8 to 10 years of age. METHODS: Sixty-six children age 8 to 10 years were examined. Convergence was evaluated using (1) nonaccommodative target at near and distance, (2) a near computerized stereogram, and (3) measurement of the near point of convergence (NPC). Reading ability was examined by (1) a reading comprehension test in which children had to answer questions regarding a paragraph they read and (2) the Developmental Eye Movement Test (DEM), which evaluates saccadic speed and accuracy. Asthenopic symptoms were scored by an Asthenopic Symptoms Questionnaire. RESULTS: Asthenopic symptoms score was correlated with the near point of convergence (r = -0.4; P = 0.003), convergence on a near stereogram (r = 0.38; P = 0.01) and distant light (r = 0.27; P = 0.04), but not with convergence on a near nonaccommodative target (r = 0.07; P = 0.6). The DEM ratio score was correlated with the asthenopic symptoms score (r = -0.32; P = 0.01), but the reading comprehension test score was not (r = 0.12; P = 0.4). There was correlation, however, between the time for completion of the reading comprehension test and the asthenopic symptoms score (r = 0.39; P = 0.006). CONCLUSION: Asthenopic symptoms score was correlated with convergence amplitude as measured, whereas accommodation is controlled and the ratio score calculated based upon DEM results. Further study is needed to evaluate the usefulness of the integration between symptom survey and objective reading examinations as screening tool for the diagnosis of convergence insufficiency. PMID- 20004876 TI - Comparison of anterior chamber depth measurements using Galilei, HR Pentacam, and Orbscan II. AB - BACKGROUND: Anterior chamber depth measurement is used frequently in cataract and refractive surgery for advanced intraocular lens (IOL) power calculation formulas and implantation of phakic IOLs. In this article, we show agreement in anterior chamber depth measurements in normal eyes among 3 noncontact systems, namely Galilei, HR Pentacam, and Orbscan II. METHODS: The anterior chamber depth measurements were obtained with Galilei, HR Pentacam, and the Orbscan II in both eyes of 37 healthy subjects. Pearson's correlations were calculated, and mean anterior chamber depth measurements were compared. Bland-Altman plots were used to assess the difference between individual measurements for each patient. RESULTS: The mean anterior chamber depth readings (+/- standard error of means) with Galilei, Pentacam, and Orbscan were 3.22 +/- 0.05, 3.25 +/- 0.05, and 3.54 +/- 0.07, respectively. The difference between Galilei and Pentacam was not statistically significant (P = 0.013). However, Orbscan measurements were larger than those of either Galilei or Pentacam (P < 0.001). The differences between Orbscan and either Galilei or Pentacam were increased with greater anterior chamber depths. The 95% limits of agreement between Orbscan and Galilei, Orbscan and Pentacam, and Pentacam and Galilei in different ranges of chamber depth were ([ < 3 mm, -0.43 to +0.71 mm]; [3 to 3.5 mm, -0.23 to +0.83]; [ > 3.5 mm, +0.41 to +0.53]), ([ < 3 mm, -0.43 to +0.71 mm]; [3 to 3.5 mm, -0.26 to +0.80]; [ > 3.5 mm, +0.38 to +0.54]), and ([ < 3 mm, -0.05 to +0.06 mm]; [3 to 3.5 mm, -0.06 to +0.10]; [ > 3.5 mm, -0.11 to +0.12]), respectively. CONCLUSIONS: These data indicate that in the assessment of normal eyes, Orbscan gives consistently higher measurements for anterior chamber depth compared with Galilei and Pentacam. Because the differences between Orbscan and either Galilei or Pentacam were not within clinically acceptable levels, they are not interchangeable in every clinical situation. However, the differences among anterior chamber depth values measured with Galilei and Pentacam were within clinically acceptable levels. Thus, these 2 systems can be regarded as interchangeable. PMID- 20004877 TI - Subjective comparison of 2 daily disposable contact lenses: Focus Dailies with AquaRelease and Proclear 1-Day. AB - BACKGROUND: Focus Dailies with AquaRelease (CIBA Vision, Duluth, Georgia) is the daily disposable contact lens currently with the highest market share. Proclear 1 Day (CooperVision, Irvine, California) is a new introduction into the daily disposable market. This study compares the subjective preference of these 2 lenses in an independent, head-to-head comparison. METHODS: Forty-one subjects were fit successfully with both brands of daily disposable contact lenses. Subjects wore the lenses for 10 consecutive days. The subjects were surveyed after the 10 days of wear to determine which lens was preferred in terms of comfort, handling, and overall preference. RESULTS: Surveys were collected from 40 subjects. Twenty-four of 40 chose Proclear 1-Day as more comfortable at the end of the day (P = 0.0003). Nineteen of 40 chose Proclear 1-Day as their overall preference (P = 0.0364). CONCLUSION: This study found that Proclear 1-Day was subjectively preferred for end-of-the day comfort and overall preference when compared with the Focus Dailies with AquaRelease or no preference. The authors hope this will give insight to eye care practitioners when deciding which daily disposable contact lens to fit. PMID- 20004878 TI - Rhino-orbito-cerebral mucormycosis, a retrospective study of 7 cases. AB - Mucormycosis is an opportunistic fungal infection caused by fungi of the Mucorales order. It has a low incidence and is a potentially lethal infection which generally affects patients who are immunocompromised due to systemic disease. We report 7 cases of rhinocerebral mucormycosis in a retrospective study of 8 years (2000-2008) in haematological patients. An early diagnosis is essential, and therefore there must be a high level of clinical suspicion in patients with predisposing factors. Certainty diagnosis requires fungal cultures or biopsies of the affected areas which prove an invasion of the tissues by the characteristic hyphae. The key to treatment is early and aggressive surgical treatment, together with high intravenous doses of amphotericin B. Despite this, prognosis is poor and mortality is about 70-80%. PMID- 20004879 TI - Epidemiology of epistaxes admitted to a tertiary hospital. AB - INTRODUCTION AND OBJECTIVES: Epistaxis has been known since antiquity. However, we have limited epidemiological data at our disposal. The aim of this study is to know the main epidemiological characteristics of the cases of epistaxis admitted and to determine the factors associated with the recurrence of bleeding. METHODS: A retrospective study was conducted including admissions for epistaxis in the ENT department of our hospital during the period between January, 2003, and December, 2008. We analyzed the distribution by gender, age, location and time of year. The aetiological causes identified included systemic and local factors. We analyzed the variables related to bleeding recurrence. RESULTS: We evaluated 178 cases of epistaxis: 68% of patients were male (121/178), compared to 32% of women (57/178). The median age (p25-p75) was 65 (53-75) years. The epistaxes were most noticeable during the months of January and April. Among the systemic causes, hypertension (56%), anti-platelet treatment (23%) and anti-coagulant therapies (18.5%) predominated. Local factors were much less numerous than general ones (11% v. 68%). Recurrent bleeding was present in 14% of cases (25/178) and only the posterior location was shown to influence this variable (P<.05). CONCLUSIONS: The typical pattern of the patients admitted to our department for epistaxis is a middle-aged or elderly male with underlying co-morbidity and posterior epistaxis. PMID- 20004880 TI - Electrowetting: a versatile tool for drop manipulation, generation, and characterization. AB - Electrowetting is arguably the most flexible tool to control and vary the wettability of solid surfaces by an external control parameter. In this article we briefly discuss the physical origin of the electrowetting effect and subsequently present a number of approaches for selected novel applications. Specifically, we will discuss the use of EW as a tool to extract materials properties such as interfacial tensions and elastic properties of drops. We will describe some modifications of the EW equation that apply at finite AC voltage for low conductivity fluids when the electric field can partially penetrate into the drops. We will discuss two examples where finite conductivity effects have important consequences, namely electrowetting of topographically structured surfaces as well as the generation of drops in AC electric fields. Finally, we review recent attempts to incorporate electrowetting into conventional channel based microfluidic devices in order to enhance the flexibility of controlling the generation of drops. PMID- 20004881 TI - Targeted next-generation sequencing appoints c16orf57 as clericuzio-type poikiloderma with neutropenia gene. AB - Next-generation sequencing is a straightforward tool for the identification of disease genes in extended genomic regions. Autozygosity mapping was performed on a five-generation inbred Italian family with three siblings affected with Clericuzio-type poikiloderma with neutropenia (PN [MIM %604173]), a rare autosomal-recessive genodermatosis characterised by poikiloderma, pachyonychia, and chronic neutropenia. The siblings were initially diagnosed as affected with Rothmund-Thomson syndrome (RTS [MIM #268400]), with which PN shows phenotypic overlap. Linkage analysis on all living subjects of the family identified a large 16q region inherited identically by descent (IBD) in all affected family members. Deep sequencing of this 3.4 Mb region previously enriched with array capture revealed a homozygous c.504-2 A>C mismatch in all affected siblings. The mutation destroys the invariant AG acceptor site of intron 4 of the evolutionarily conserved C16orf57 gene. Two distinct deleterious mutations (c.502A>G and c.666_676+1del12) identified in an unrelated PN patient confirmed that the C16orf57 gene is responsible for PN. The function of the predicted C16orf57 gene is unknown, but its product has been shown to be interconnected to RECQL4 protein via SMAD4 proteins. The unravelled clinical and genetic identity of PN allows patients to undergo genetic testing and follow-up. PMID- 20004882 TI - Maternal and newborn morbidity by birth facility among selected United States 2006 low-risk births. AB - OBJECTIVE: We sought to evaluate perinatal morbidity by delivery location (hospital, freestanding birth center, and home). STUDY DESIGN: Selected 2006 US birth certificate data were accessed online from the Centers for Disease Control and Prevention. Low-risk maternal and newborn outcomes were tabulated and compared by birth facility. RESULTS: A total of 745,690 deliveries were included, of which 733,143 (97.0%) occurred in hospital, 4661 (0.6%) at birth centers, and 7427 (0.9%) at home. Compared with hospital deliveries, home and birthing center deliveries were associated with more frequent prolonged and precipitous labors. Home births experienced more frequent 5-minute Apgar scores <7. In contrast, home and birthing center deliveries were associated with less frequent chorioamnionitis, fetal intolerance of labor, meconium staining, assisted ventilation, neonatal intensive care unit admission, and birthweight <2500 g. CONCLUSION: Home births are associated with a number of less frequent adverse perinatal outcomes at the expense of more frequent abnormal labors and low 5 minute Apgar scores. PMID- 20004883 TI - Counseling and screening for chromosomal abnormalities. PMID- 20004884 TI - Continuous infusion of 17-hydroxyprogesterone caproate into either the fetoplacental or intervillous circulation of a placental cotyledon attenuates vasoconstriction of the fetoplacental arteries by thromboxane mimetic U46619. AB - OBJECTIVE: The objective of the study was to determine whether pretreatment of fetal or maternal placental vasculature with 17-hydroxyprogesterone caproate (17 P) attenuates the vasoactive effect of the thromboxane mimetic U46619. STUDY DESIGN: Two cotyledons were obtained from each placenta studied. For the first 5 placentas, the fetal artery of 1 cotyledon from each pair was infused with 17-P. After 30 minutes, a bolus dose of U46619 was administered to both cotyledons. An identical procedure was carried out on the next 5 placentas except that 17-P was infused into the intervillous space. RESULTS: The pressure excursion caused by bolus administration of U46619 was less in the cotyledons infused with 17-P, both in the 5 cases in which the fetal vasculature was infused with 17-P (P = .0035) and in the 5 cases in which the maternal vasculature was infused with 17-P (P = .038). CONCLUSION: Pretreatment of either the fetal or maternal circuits of the placenta with 17-P attenuates U46619-mediated fetoplacental vasoconstriction. PMID- 20004886 TI - The natural history of cervical and vulvar intraepithelial neoplasia. PMID- 20004887 TI - [Electrical storm secondary to hypokaliemia in a patient with implantable cardioverter defibrillator]. AB - Electrical storm in patients with implantable cardioverter defibrillator (ICDs) is a rhythmic emergency which can be immediately lethal. It occurs especially in patients with an advanced cardiomyopathy. Such arrhythmias predictor factors and triggers are rarely found. We report the case of a 73-year-old man with ischemic dilated cardiomyopathy, who underwent a complete surgical revascularisation six years ago. In 2003, this patient was hospitalised in cardiology because of a sustained ventricular tachycardia reduced by electrical shock. Because of the aggravation of the heart failure with a NYHA functional class III, electrical and echocardiographic criteria of ventricular dyssynchrony, a biventricular ICD was implanted. Three years after, the patient was hospitalized because of an electrical storm with 96 appropriate shocks. A severe hypokaliemia was the cause of this electrical storm, and the evolution was favourable after correction of the hypokaliemia. Hypokaliemia is rarely the trigger of such arrhythmias, it represents only 3 % of the causes. Its prevalence may be underestimated especially in patients with heart failure who receive high doses of diuretics. PMID- 20004888 TI - Sleep, illness course, and concurrent symptoms in inter-episode bipolar disorder. AB - We investigated associations between sleep, illness course, and concurrent symptoms in 21 participants with bipolar disorder who were inter-episode. Sleep was assessed using a week-long diary. Illness course and symptoms were assessed via validated semi-structured interviews. Lower and more variable sleep efficiency and more variable total wake time were associated with more lifetime depressive episodes. Variability in falling asleep time was positively correlated with concurrent depressive symptoms. Sleep efficiency was positively correlated with concurrent manic symptoms. These findings suggest that inter-episode sleep disturbance is associated with illness course and that sleep may be an important intervention target in bipolar disorder. PMID- 20004889 TI - Facile synthesis of D-lividosamine. AB - A simple, four-step synthesis of D-lividosamine starting from N-acetyl-D glucosamine via a furanosyl oxazoline intermediate is described. PMID- 20004890 TI - 1,5-Anhydro-D-fructose: biocatalytic and chemical synthetic methods for the preparation, transformation and derivatization. AB - 1,5-Anhydro-D-fructose (1,5AnFru) is a monoketosaccharide that can be prepared enzymatically from starch by alpha-1,4-glucan lyase or chemically from D-glucose or D-fructose in a few steps with high yields. The formed 1,5AnFru can be derivatized both enzymatically and chemically to interesting new carbohydrate derivatives, some with biological activities. For example dehydratases, isomerases and reductases can convert 1,5AnFru to enolones (as Ascopyrone P) and sugar alcohols with antimicrobial and antioxidant properties, while chemical modifications can give similar compounds as well as natural products like 1 deoxymannonojirimycin and Clavulazine. 1,5AnFru disaccharides (glycosyl 1-->4 1,5AnFru) have been prepared as well as glycosyl 1-->4 1,5-anhydro-D-tagatose. PMID- 20004891 TI - Development of new glycosylation methodologies for the synthesis of archaeal derived glycolipid adjuvants. AB - To commercialize the production of glycolipid adjuvants, their synthesis needs to be both robust and inexpensive. Herein we describe a semi-synthetic approach where the lipid acceptor is derived from the biomass of the archaeon Halobacterium salinarum, and the glycosyl donors are chemically synthesized. This work presents some preliminary results using the promoter system N iodosuccinimide (NIS) and a stable 0.25 M solution of boron trifluoride trifluoroethanol (BF(3) x TFE(2)) in dichloromethane. This promoter system allows for the use of peracetyl alkyl(aryl)thioglycosides that are available in high yield from inexpensive disaccharide starting materials by promoting clean glycosylation reactions from which pure product can be easily isolated. Conventional glycosylation using NIS-silver trifluoromethanesulfonate (AgOTf) leads to extensive acetyl transfer to the archaeol acceptor and numerous byproducts that make purification complicated. As part of preliminary structure adjuvant activity studies, we describe the one-pot synthesis of a gentiobiose beta-Glcp-(1-->6)-Glcp-SEt donor with an O-2 benzoyl group, which can be used to prepare a disaccharide attached to archaeol in 85% overall yield, and the related glycolipid trisaccharide beta-Glcp-(1-->6)-beta-Glcp-(1-->6)-beta-Glcp-(1-->O) archaeol. The synthesis of the isomeric beta-Glcp-(1-->6)-alpha-Glcp-(1-->O) archaeol featuring a >10:1 alpha/beta alpha-selective glycosylation using the promoter system N-phenylselenylphthalimide-trifluoromethanesulfonic acid (TfOH) is also presented, along with the adjuvant properties of the corresponding archaeosomes (liposomes comprised entirely of combinations of isoprenoid archaeal like lipids). These new vaccine formulations extend previous observations that glycolipids are integral to the activation of MHC type I pathways via CD8(+) antigen-specific T-cells. The beta-Glcp-(1-->6)-beta-Glcp-(1-->6)-beta-Glcp-(1- >O)-archaeol trisaccharide is shown to be more active than the Glcp-(1-->6)-beta Glcp-(1-->O)-archaeol disaccharide. PMID- 20004892 TI - Influence of patient age on the association between euploidy and day-3 embryo morphology. AB - A retrospective cohort study conducted in 138 patients undergoing their first preimplantation genetic screening (PGS) cycle between January 1, 2006, and December 31, 2007, demonstrated that embryos with good day-3 morphology were more likely to be euploid for X/Y, 8, 15, and 18 than those with poor morphology. The strength of association between euploidy and day-3 morphology was not influenced by maternal age. PMID- 20004893 TI - Transgastric endoluminal gastrojejunostomy: technical development from bench to animal study (with video). AB - BACKGROUND: Gastrojejunal anastomosis is commonly performed for palliative management of malignant gastric outlet obstruction and bariatric surgery. Natural orifice transluminal endoscopic surgery revolutionized the surgical approach to intra-abdominal surgery. This study explored the possibility of performing gastrojejunostomy (GJ) by using a hybrid natural orifice transluminal endoscopic surgery approach. OBJECTIVE: To develop a surgical technique for the performance of transgastric endoscopic GJ (TGEJ) in a porcine model. DESIGN: Prospective series of animal experiments. SETTING: University hospital animal laboratory. ANIMALS: Thirteen female domestic pigs. INTERVENTIONS: With the animals under general anesthesia, the endoscope is passed through the gastrotomy and a segment of small bowel is retrieved into the stomach. An enterotomy is then created, and an EndoGIA stapler is introduced through an intragastric port and passed between the small bowel and stomach wall. A GJ is formed after firing of the EndoGIA stapler. The pigs are allowed to resume their diet 1 day after the operation and are allowed to survive for 2 weeks before they are euthanized. The patency of the GJ is confirmed with a repeat endoscopy, contrast study, and postmortem examination. RESULTS: A total of 13 TEGJs were performed, 11 of which were successful. The mean operative time was 53.6 +/- 45.7 minutes. The mean time for gastrotomy was 4.7 minutes, and that for GJ was 42.5 minutes. One TEGJ was converted to open surgery because of malpositioning of the intragastric port, and the other failed because the enterotomy was too extensive. Ten of 11 pigs survived for 2 weeks, and endoscopic examination with contrast study confirmed that all the gastrojejunostomies were patent. On postmortem examination, the average size of the GJ was 30 mm. LIMITATIONS: The length between duodenojejunal flexure and the site chosen to perform the GJ could not be determined. CONCLUSIONS: TEGJ is technically feasible with a patent and sizable anastomosis. PMID- 20004894 TI - Postoperative surgical site infection following acetabular fracture fixation. AB - Postoperative surgical site infection (SSI) in orthopaedic trauma surgery is uncommon, but can present serious complications. This study was designed to assess the prevalence of, and to identify the risk factors for, SSI following acetabular fracture open reduction and internal fixation. A total of 326 consecutive patients who underwent acetabular fracture surgery were retrospectively reviewed. There were 17 patients (5.2%) who developed a SSI, including 10 deep infections and 7 superficial infections. Staphylococcus aureus was the most common causative pathogens in 9 patients, and was Methicillin resistant in 3 patients. Enterococcus faecalis was found in 6 patients, Staphylococcus epidermidis in 3 patients, and Pseudomonas aeruginosa and enterbacter cloacae in 2 patients each. Fourteen of 17 patients developed their infection within 4 weeks after the fixation. Univariate analysis demonstrated that the SSI group had statistically significant higher Injury Severity Score, longer intensive care unit (ICU) stays, larger amount of packed red blood cells transfused, longer operative time, larger estimated operative blood loss, higher body mass index (BMI), more frequent performance of combined approach, embolisation of internal iliac arteries, association of urinary tract injury, and Morel-Lavallee lesion compared to the no SSI group. Multivariate analysis using these 10 parameters showed that BMI, ICU stay, and Morel-Lavallee lesion were independently significant risk factors for SSI. To reduce the incidence of SSI following acetabular fracture surgery, special attention should be directed at the care of obese patients, patients requiring ICU care, and patients with associated Morel-Lavallee lesions. PMID- 20004895 TI - The metabolic syndrome adds utility to the prediction of mortality over its components: The Vietnam Experience Study. AB - BACKGROUND: The metabolic syndrome increases mortality risk. However, as "non affected" individuals may still have up to two risk factors, the utility of using three or more components to identify the syndrome, and its predictive advantage over individual components have yet to be determined. METHODS: Participants, male Vietnam-era veterans (n=4265) from the USA, were followed-up from 1985/1986 for 14.7 years (61,498 person-years), and all-cause and cardiovascular disease deaths collated. Cox's proportional-hazards regression was used to assess the effect of the metabolic syndrome and its components on mortality adjusting for a wide range of potential confounders. RESULTS: At baseline, 752 participants (17.9%) were identified as having metabolic syndrome. There were 231 (5.5%) deaths from all causes, with 60 from cardiovascular disease. After adjustment for a range of covariates, the metabolic syndrome increased the risk of all-cause, HR 2.03, 95%CI 1.52, 2.71, and cardiovascular disease mortality, HR 1.92, 95%CI 1.10, 3.36. Risk increased dose-dependently with increasing numbers of components. The increased risk from possessing only one or two components was not statistically significant. The adjusted risk for four or more components was greater than for only three components for both all-cause, HR 2.30, 95%CI 1.45, 3.66 vs. HR 1.70, 95%CI 1.11, 2.61, and cardiovascular disease mortality, HR 3.34, 95%CI 1.19, 9.37 vs. HR 2.81, 95%CI 1.07, 7.35. The syndrome was more informative than the individual components for all-cause mortality, but could not be assessed for cardiovascular disease mortality due to multicollinearity. Hyperglycaemia was the individual strongest parameter associated with mortality. CONCLUSIONS: The metabolic syndrome is informative in predicting mortality, with risk increasing as the number of components increase above the threshold required for diagnosis. PMID- 20004896 TI - Editor's report. PMID- 20004897 TI - Osteocalcin positive mononuclear cells are associated with the severity of aortic calcification. AB - OBJECTIVE: To assess the association of circulating bone marrow-derived osteo progenitors with vascular calcification in mouse models and patients with peripheral artery disease. METHODS: We estimated the percentage of circulating mononuclear cells expressing osteocalcin in 2 mouse models of aortic calcification developed in osteoprotegerin-deficient mice (OPG(-/-)) using flow cytometry. Aortic calcification was assessed in mice principally by a bioassay of harvested aortas. In patients with peripheral artery disease osteocalcin-positive cells (estimated by flow cytometry) were related to aortic calcification volume assessed from computed tomography. RESULTS: The amount of extractable aortic calcium was increased in both mouse models used in comparison to controls. The percentage of circulating mononuclear cells expressing osteocalcin was correlated to the amount of extractable aortic calcium in male (r=0.525, p=0.02) and female OPG(-/-) (r=0.564, p=0.02) mice and also in animals in which calcification was accelerated using calcitriol (r=0.64, p=0.01). Patients with more severe aortic calcification had a greater percentage of circulating OCN(+) MNCs (median 4.07%, IQR 3.76-4.39, n=12) than those with less severe aortic calcification (median 3.10%, IQR 2.32-3.60, n=11, p=0.05). CONCLUSIONS: This study demonstrates that aortic calcification can be robustly quantified in 2 mouse models. In these models and patients with peripheral artery disease circulating osteocalcin positive mononuclear cells are associated with the severity of aortic calcification. PMID- 20004898 TI - Changes in force and tendinous tissue elongation during the early phase of tetanic summation in in vivo human tibialis anterior muscle. AB - The purpose of this study was to determine the changes that occur in tendinous tissue properties during the early phase of tetanic summation in the in vivo human tibialis anterior muscle (TA). The torque response and tendinous tissue elongation following single stimuli, two-pulse trains, and three-pulse trains were recorded in the TA during isometric contractions. The elongation, compliance, and lengthening velocity of tendinous tissue were determined by real time ultrasonography. The contribution of the response to the second stimulation (C2) was obtained by subtracting the response to the single stimulation (C1) from the response of doublet. The third contribution (C3) was obtained by subtracting the response to the doublet from that of the triplet. C2 (7.8+/-0.5 Nm) and C3 (7.3+/-0.6 Nm) had torque responses significantly higher than C1 (3.6+/-0.7 Nm). In contrast, the elongations of tendinous tissue for C2 (2.8+/-0.4mm) and C3 (1.7+/-0.2mm) were significantly lower than for C1 (4.9+/-0.3mm), indicating that the summation pattern of tendinous tissue elongation is different from the summation pattern of torque response. In addition, this showed considerable difference both between C1 (0.12+/-0.01 mm/N; 83+/-4.6mm/s) and C2 (0.03+/-0.005 mm/N; 50+/-6.3mm/s) and between C1 and C3 (0.02+/-0.002 mm/N; 39+/-6.4mm/s) in the compliance and lengthening velocity of tendinous tissue. These results suggest that changes in tendinous tissue properties between first and second contraction are related to different summation patterns of force and tendinous tissue elongation during early phase of tetanic summation. PMID- 20004899 TI - Detecting femur-insert collisions to improve precision of fluoroscopic knee arthroplasty analysis. AB - Fluoroscopic analysis is an important tool for assessing in vivo kinematics of knee prostheses. Most commonly, a single-plane fluoroscopic setup is used to capture the motion of prostheses during a particular task. Unfortunately, single plane fluoroscopic analysis is imprecise in the out-of-plane direction. This can result in reconstructing physically impossible poses, in which--for example--the femoral component intersects with the insert, as the normal pose estimation process does not take into account the relation between the components. In the proposed method, the poses of both components are estimated simultaneously, while preventing femur-insert collisions. In a phantom study, the accuracy and precision of the new method in estimating the relative pose of the femoral component were compared to those of the original method. With reverse engineered models, the errors in estimating the out-of-plane position decreased from 2.0+/ 0.7 to 0.1+/-0.1 mm, without effects on the errors in rotations and the in-plane positions. With CAD models, the errors in estimating the out-of-plane position decreased from 5.3+/-0.7 mm (mean+/-SD) to 0.0+/-0.4 mm, at the expense of a decreased precision for the other position or orientation parameters. In conclusion, collision detection can prevent reconstructing impossible poses and it improves the position and motion estimation in the out-of-plane direction. PMID- 20004900 TI - Strain shielding in distal femur after patellofemoral arthroplasty under different activity conditions. AB - Strain shielding, a mechanical effect occurring in structures combining stiff with more flexible materials, is considered to lead to a reduction of density in bone surrounding the implant. This effect can be related to the weakness of the implant fixation, which can promote implant loosening. Several studies describe a significant decrease in postoperative bone mineral density adjacent to joint implants, which can compromise their long-term fixation. The aim of the present study was to quantify the strain shielding effect on the distal femur after patellofemoral arthroplasty. For this purpose three activities of daily living were considered: level walking, stair climbing and deep bending at different angles of knee flexion. To determine the strain shielding effect, cortical bone strains were measured experimentally with triaxial strain gauges in synthetic femurs before and after patellofemoral arthroplasty for each of the different daily activities. The results showed that the patellofemoral arthroplasty in general reduced the strains in the medial and distal regions of the femur when deep bending activity occurred, consequently, strain shielding in these regions, with strain decreases of -72.0% and -67.5% were measured. On the other side, higher values of strain were found in the anterior region after patellofemoral replacement for this activity with an increase of +182.0%. The occurrence of strain shielding seems to be more significant when the angle of knee flexion and applied load increases. Strain shielding and over-loading may have relevant effects on bone remodeling surrounding the patellofemoral implant, suggesting a potential effect of later bone resorption in the medial and distal femur regions in case of regular deep bending activity. PMID- 20004901 TI - Determination of pyridine, 2-picoline, 4-picoline and quinoline from mainstream cigarette smoke by solid-phase extraction liquid chromatography/electrospray ionization tandem mass spectrometry. AB - The present paper describes the development and validation of a new reversed phase liquid chromatography-electrospray ionization tandem mass spectrometric method (RP-HPLC-ESI-MS/MS) for simultaneous determination of pyridine, 2 picoline, 4-picoline and quinoline from mainstream cigarette smoke. Liquid-liquid extraction followed by solid-phase extraction was applied to extract the target analytes from cigarette smoke. Baseline chromatographic separation was achieved by utilizing a Zorbax SB-Aq (4.6x150 mm, 5 microm) column in gradient chromatographic conditions with acetonitrile and ammonium acetate buffer as mobile phases. Popular commercially available Indian brand filtered and non filtered cigarettes were analyzed using the same method. The identification of each chemical was established by chromatographic retention times, analyte specific fragmentation patterns and relative peak area ratios of two product/precursor ion pairs. The limit of detection of this method ranged from 1.74 to 14.32 ng/cig using an injection volume of 20 microl. The reproducibility of this method is excellent and better standard deviations were obtained compared to literature reported values for these chemicals. RSD value is less than 9% for all analytes. PMID- 20004902 TI - Analysis of part-per-billion level of arsine and phosphine in light hydrocarbons by capillary flow technology and dielectric barrier discharge detector. AB - A practical gas chromatographic procedure has been developed and implemented for the measurement of arsine and phosphine in hydrocarbons such as propylene at the part-per-billion level. The successful measurement of arsine and phosphine at the level mentioned was attained by incorporating a large volume injection technique to increase the mass of solutes delivered for sensitivity improvement, capillary flow technology to keep the matrix from entering the detector by either back flushing through the inlet vent, or by heart-cutting if required, and dielectric barrier discharge detector operating in argon mode for sensitivity enhancement, as well as offering improved selectivity towards the solutes cited. Using the technique described a complete analysis can be conducted in less than 4 min. A relative standard precision of less than 1.7% was achieved with repeated injections at the concentration level of 25 and 125 ppb (v/v) each of arsine and phosphine in nitrogen with a practical detection limit at the 5 ppb (v/v) level. Correlation coefficients of greater than 0.9999 were obtained for arsine and phosphine over a range from 10 to 2500 ppb (v/v). The analytical methodology was proven to be reliable in continuous operation during the first 6 months of deployment. PMID- 20004903 TI - Insights into the mechanism of separation of heparin and heparan sulfate disaccharides by reverse-phase ion-pair chromatography. AB - Reverse-phase ion-pair high performance liquid chromatography (RPIP-HPLC) and ultra-performance liquid chromatography (RPIP-UPLC) are increasingly popular chromatographic techniques for the separation of organic compounds. However, the fine details of the RPIP separation mechanism are still being debated. Many factors including type and concentration of the ion-pairing reagent, mobile phase pH, organic modifier, ionic strength, and stationary phase all play a role in the overall efficiency and optimization of ion-pairing separations. This study investigates the role that competition between ion-pairing reagents with different steric bulk and hydrophobicity plays in the separation of structural isomers of heparin and heparan sulfate (HS) disaccharides. In addition to providing insights into the mechanism by which RPIP-HPLC can resolve closely related disaccharides, the use of competition between ion-pairing agents could lead to new methods for the separation of larger heparin and HS oligosaccharides. This approach should also be applicable to the analysis of other compound classes, and could lead to a general approach for the chromatographic resolution of mixtures of charged analytes having similar structures. PMID- 20004904 TI - Spectroscopic analysis of interactions between alkylated silanes and alumina nanoporous membranes. AB - Transport across alumina nanoporous membranes can be altered via surface attachment of alkylated trimethoxysilane compounds. The mechanism of attachment has been previously assumed to be monolayer silane coverage through full chemisorption regardless of reaction conditions. This chemisorption arises via covalent Si-O-Al bond formation resulting from condensation between the three putative silanols (due to hydrolysis of the three Si-OCH(3) bonds) and hydroxides present on the alumina surface. If this model was correct, methanol would be produced in large quantities in the reaction solution, and the methoxy moieties would no longer be present on the silane molecule. The results presented in this paper utilized FT-IR and both solution and solid-state NMR to examine the chemical nature of octadecyltrimethoxysilane (ODTMS) present on the alumina surface. The FT-IR results confirm the presence of the silane on the membrane. The (1)H solution NMR results indicate small but detectable methanol production during attachment. The solid-state NMR results demonstrate that the methoxy proton NMR integrated peak intensities remain in nearly the same ratios present in the free silane, concluding that the majority of methoxy groups are intact while the silane is attached to the membrane surface. These three results suggest that monolayer surface coverage and chemisorption through full covalent bonding is not the primary means of attachment for ODTMS on the surface of alumina nanomembranes under these reaction conditions. PMID- 20004905 TI - Porosity evolution of VP-DVB/MCM-41 nanocomposite. AB - The porous structure of nanostructured vinylpyrrolidone-divinylbenzene (VP DVB)/MCM-41 composite was characterized using N(2) adsorption at 77K and positronium annihilation lifetime spectroscopy (PALS), atomic force microscopy (AFM) and Raman spectroscopy. Positron annihilation lifetime spectra were measured during outgassing procedure. The voids of differentiated dimensions were detected in the composite material. The number of free volumes and their dimensions depend on the degree of evacuation of volatile components from the samples. PMID- 20004906 TI - Biosorption of nickel onto treated alga (Oedogonium hatei): Application of isotherm and kinetic models. AB - Oedogonium hatei was developed into an effective and efficient adsorbent for the removal of Ni(II) ions from aqueous solution. The adsorption studies of untreated and treated algal biomass (with 0.1M HCl) were compared in batch mode. Optimum biosorption conditions were determined as a function of pH, biomass dosage, contact time, and temperature and the Langmuir and Freundlich isotherms were found applicable in terms of relatively high regression values. The maximum monolayer adsorption capacity of the biosorbents (untreated and acid-treated algae), as obtained from the Langmuir adsorption isotherm, was found to be 40.9 and 44.2mg/g, respectively at 80min contact time, 5.0 pH, 0.7g/L algal dose, and 298K temperature. The thermodynamic parameters showed that the adsorption of Ni(II) ions onto algal biomass was feasible, spontaneous, and exothermic under the studied conditions. Kinetics of adsorption followed both first- and second order rate equations and the process involving the rate-controlling step is complex involving boundary layer as well as intraparticle diffusion processes. The FTIR results of algal biomass showed that biomass has different functional groups and these functional groups are able to react with metal ion in aqueous solution. Biosorbent could be regenerated using 0.1M NaOH solution, with up to 70% recovery. The performance of this biosorbent was then compared with many other reported biosorbents for nickel removal and it was observed that the proposed adsorbent is effective in terms of its performance. PMID- 20004907 TI - Aqueous foam stabilized by plate-like particles in the presence of sodium butyrate. AB - The addition of salt promotes the adsorption of layered double hydroxide (LDH) particles onto the air-water interface, but stable foams cannot be prepared from LDH dispersions at all the concentration of NaCl or sodium acetate. We generated stable foams using positively charged plate-like LDH particles in the presence of sodium butyrate. The effects of adding sodium butyrate to LDH on the particle zeta potential, adsorption behavior and the adsorption of modified particles at the air-water interface were studied. At a fixed LDH particle concentration, adding of a trace amount of sodium butyrate maximizes flocculation of the aqueous particle dispersion. Foams prepared under this condition of particle dispersion are most stable to coalescence and halt completely disproportionation. Also, the size of the bubbles is the smallest. The bubbles are stable when drying at 80 degrees C with little change in size. Laser-induced fluorescent confocal micrographs and scanning electron microscopy observations clearly confirm the adsorption of LDH particles on the foam surfaces, and the bubbles are armored by an interfacial particle multilayer. PMID- 20004908 TI - Word learning by children with phonological delays: differentiating effects of phonotactic probability and neighborhood density. AB - This study examined the ability of 20 preschool children with functional phonological delays and 34 age- and vocabulary-matched typical children to learn words differing in phonotactic probability (i.e., the likelihood of occurrence of a sound sequence) and neighborhood density (i.e., the number of words that differ from a target by one phoneme). Children were exposed to nonwords paired with novel objects in a story and learning was measured by a picture naming task. Results showed that both groups created lexical representations for rare sound sequences from sparse neighborhoods. However, only children with typical development appeared to build on this initial lexical representation to create a full representation of the word (i.e., lexical-semantic connection and semantic representation). It was hypothesized that creating a lexical representation may be too resource demanding for children with phonological delays, leaving few resources available to create a lexical-semantic connection and/or a semantic representation. LEARNING OUTCOMES: The reader will be able to (1) define phonotactic probability; (2) define neighborhood density; (3) identify how these variables impact the word learning process in general; (4) identify potential areas of deficit in the word learning process for children with functional phonological delays. PMID- 20004909 TI - Learning of syllable-object relations by preverbal infants: the role of temporal synchrony and syllable distinctiveness. AB - The role of temporal synchrony and syllable distinctiveness in preverbal infants' learning of word-object relations was investigated. In Experiment 1, 7- and 8 month-olds (N=64) were habituated under conditions where two similar-sounding syllables, /tah/ and /gah/, were spoken simultaneously with the motions of one of two sets of objects (synchronous) or out of phase with the motions (asynchronous). On test trials, 8-month-olds, but not 7-month-olds, showed learning of the relations in the synchronous condition but not in the asynchronous condition. Furthermore, in Experiment 2, following habituation to one of the synchronous syllable-object pairs, 7-month-olds (n=8) discriminated the syllables and the objects. In Experiment 3, following habituation to two distinct syllables, /tah/-/gih/ or /gah/-/tih/, paired with identical objects, 7 month-olds (n=40) showed learning of the relations, again only in the synchronous condition. Thus, synchrony, which mothers naturally provide between words and object motions, facilitated the mapping onto objects of similar-sounding syllables at 8months of age and distinct syllables at 7months of age. These findings suggest an interaction between infants' synchrony and syllable distinctiveness perception during early word mapping development. PMID- 20004910 TI - The long-term effects of breastfeeding on child and adolescent mental health: a pregnancy cohort study followed for 14 years. AB - OBJECTIVES: To determine whether there was an independent effect of breastfeeding on child and adolescent mental health. STUDY DESIGN: The Western Australian Pregnancy Cohort (Raine) Study recruited 2900 pregnant women and followed the live births for 14 years. Mental health status was assessed by the Child Behaviour Checklist (CBCL) at 2, 6, 8, 10, and 14 years. Maternal pregnancy, postnatal, and infant factors were tested in multivariable random effects models and generalized estimating equations to examine the effects of breastfeeding duration on mental health morbidity. RESULTS: Breastfeeding for less than 6 months compared with 6 months or longer was an independent predictor of mental health problems through childhood and into adolescence. This relationship was supported by the random effects models (increase in total CBCL score: 1.45; 95% confidence interval 0.59, 2.30) and generalized estimating equation models (odds ratio for CBCL morbidity: 1.33; 95% confidence interval 1.09, 1.62) showing increased behavioral problems with shorter breastfeeding duration. CONCLUSION: A shorter duration of breastfeeding may be a predictor of adverse mental health outcomes throughout the developmental trajectory of childhood and early adolescence. PMID- 20004911 TI - Dietary quality index and associated factors among adolescents of the state of Sao Paulo, Brazil. AB - OBJECTIVE: To assess dietary quality and associated factors in adolescents. STUDY DESIGN: We conducted a population-based cross-sectional study in a sample of 1584 adolescents living in areas of the state of Sao Paulo, Brazil. Dietary intake was measured with the 24-hour recall method, and dietary quality was assessed by means of the Health Eating Index (HEI), adapted to fit to the local requirements. Linear regression analyses were performed to assess the association between the HEI and demographic, socioeconomic, and lifestyle variables. RESULTS: A total of 97.1% of the adolescents studied had an inadequate diet or a diet that needed improvement. The mean overall HEI score was 59.7. Lower mean HEI scores were found for fruits, dairy products, and vegetables. Male adolescents who were physically active and lived in a house or apartment had higher HEI scores. The multiple regression analyses showed that the quality of the diet improved as age decreased. Adolescents who lived in houses or apartments had higher HEI scores than adolescents living in shacks or slums, regardless of age and energy intake. CONCLUSIONS: Dietary quality is associated with income and age. A better understanding of the factors associated can provide input to the formulation of policies and development of nutritional actions. PMID- 20004912 TI - Maternal preeclampsia predicts the development of bronchopulmonary dysplasia. AB - OBJECTIVE: To test the hypothesis that exposure to preeclampsia is associated with an increased risk of bronchopulmonary dysplasia (BPD). STUDY DESIGN: A prospective cohort study of 107 babies born between 23 and 32 weeks gestation, collecting maternal, neonatal, and placental data. RESULTS: Of the 107 infants studied, 27 (25%) developed BPD. The bivariate odds ratio (OR) for the relationship between pre-eclampsia and BPD was 2.96 (95% confidence interval [CI] = 1.17 to 7.51; P = .01). When controlling for gestational age, birth weight z score, chorioamnionitis, and other clinical confounders, the OR of developing BPD was 18.7 (95% CI = 2.44 to 144.76). Including the occurrence of preeclampsia, clinical chorioamnionitis, male sex, and maternal tobacco use in addition to gestational age and birth weight z-score accounted for 54% of the variability of the odds of developing BPD. CONCLUSIONS: BPD is increased for infants exposed to preeclampsia. This has possible implications for the prevention of BPD with proangiogenic agents, such as vascular endothelial growth factor. PMID- 20004913 TI - Adiposity predicts carotid intima-media thickness in healthy children and adolescents. AB - OBJECTIVE: To examine whether anthropometric measurements, blood pressure (BP), fasting total cholesterol, and low-density lipoprotein are related to ultrasound measures of carotid intima-media thickness (CIMT) in children and teenagers with no known risk factors for cardiovascular disease. STUDY DESIGN: This cross sectional study included 599 subjects, 6 to 20 years of age (292 males, 307 females; 224 Hispanics, 210 European-Americans, 126 African-Americans, and 39 Asian-Americans) whose body mass index, waist circumference, BP, lipid profiles, and values for CIMT were determined. RESULTS: Measures of CIMT were significantly greater in males than females (P=.006) and in African-Americans when compared with other ethnic groups (all P < .05). There were no relations between age, diastolic BP, or fasting levels of triglycerides, total cholesterol, or low density lipoprotein values and CIMT measures, regardless of sex or ethnic background. Stratified multiple regression analysis indicated that body mass index and waist circumference independently predicted CIMT in both males and females, even after controlling for age, weight, BP, fasting lipid levels, and ethnic background. CONCLUSION: Increased body mass and adiposity are associated with increased intima-media thickness in children and teenagers. This association is present in children not considered overweight, underscoring the need for the continued promotion of adequate nutritional and physical exercise behavior during childhood. PMID- 20004914 TI - Neural state and trait bases of mood-incongruent memory formation and retrieval in first-episode major depression. AB - Mood-congruent cognitive biases constitute critical factors for the vulnerability to depression and its maintenance. One important aspect is impaired memory for positive information during depression and after recovery. To elucidate its state (during depression only) and trait (during depression and recovery) related neural bases, we investigated medication free depressed, recovered, and healthy individuals with functional MRI while they memorized and recognized happy and neutral face stimuli. The imaging results revealed group differences in mood incongruent successful memory encoding and retrieval activity already in the absence of significant memory performance differences. State effects were observed in the amygdala and posterior cingulate cortex. Whereas the amygdala was generally involved in memory formation, its activity predicted subsequent forgetting of neutral faces in depressed patients. Furthermore, the amygdala and posterior cingulate cortex were involved in memory retrieval of happy faces in depressed patients only. Trait effects were observed in the fusiform gyrus and prefrontal cortex. The fusiform gyrus was involved in memory formation and retrieval of happy faces in both patient groups, whereas it was involved in memory formation and retrieval of neutral faces in healthy individuals. Similar trait effects were observed during memory retrieval in the orbitofrontal cortex and left inferior frontal gyrus. Thus, while memory processing of positive information in the amygdala and posterior cingulate cortex is biased during depression only, memory processing in the fusiform gyrus and prefrontal cortex is biased also after recovery. These distinct neural mechanisms may respectively constitute symptom maintenance and cognitive vulnerability factors for depression. PMID- 20004915 TI - A pilot study of the phase angle between cortisol and melatonin in major depression - a potential biomarker? AB - INTRODUCTION: Hypothalamic-pituitary-adrenal (HPA) axis and melatonin rhythm alterations have been independently reported in major depression (MDD) as well as in insomnia. In this pilot study, we link cortisol and melatonin rhythms and propose that the phase angle between cortisol acrophase (CA) and dim-light melatonin onset (20 pg/ml) (DLMO-20) may yield a useful state specific biomarker for MDD. METHODS: Six healthy (HC) and six depressed (MDD) psychotropic free subjects were admitted to the General Clinical Research Center. Blood was sampled for cortisol and melatonin from 1600h to 1000h, under dim lights (<20lux) and constant routine. Time for DLMO-20 and peak cortisol concentration was determined for each subject. Phase angle was computed as the difference in time between CA and DLMO-20. RESULTS: Phase angle was significantly increased in MDD's versus HC's (13.40+/-1.61h. versus 11.61+/-1.66h, p=0.026). Using ROC analysis, a phase angle greater than 13.57h distinguished MDD's from HC's (sensitivity=0.83, specificity=1.0). Mean nocturnal melatonin (1600-1000h) was significantly decreased in MDD's versus HC's (22.67+/-9.08 pg/ml versus 47.82+/-14.76 pg/ml, p=0.015). CONCLUSIONS: The phase angle between CA and DLMO-20 appears to distinguish HC's from MDD's and may be a useful biomarker to aid biologic assessment as well as treatment. Lower nocturnal melatonin in MDD's highlights its importance in MDD's pathophysiology. Additional study with larger sample size is needed to confirm the results of this pilot study. The mechanism for this phase angle difference and decreased melatonin, itself, requires further study. PMID- 20004916 TI - The use of spirometry testing prior to cardiac surgery may impact the Society of Thoracic Surgeons risk prediction score: a prospective study in a cohort of patients at high risk for chronic lung disease. AB - OBJECTIVES: Chronic lung disease is a significant comorbidity in patients undergoing cardiac surgery. Chronic lung disease is currently being classified and reported to the Society of Thoracic Surgeons database by using either clinical interview or spirometric testing. We sought to compare the chronic lung disease classification captured by the 2 methods. METHODS: We performed a prospectively designed study in which patients presenting for cardiac surgery, excluding emergent patients, were screened for a history of asthma, a history of 10 or more pack-years of smoking, a persistent cough, and the use of oxygen. Each selected patient underwent spirometry. The presence and severity of chronic lung disease was coded per Society of Thoracic Surgeons guidelines by using the 2 methods of clinical report and spirometric results. The chronic lung disease classifications were compared, and differences were determined by using concordance and discordance rates. The results were then used to construct Society of Thoracic Surgeons-predicted risk models. RESULTS: The discordant rate was 39.1%, with underestimation of the severity of chronic lung disease in 94% of misclassified patients. This affected the Society of Thoracic Surgeons-predicted risk models for prolonged ventilation, morbidity/mortality, and mortality by increasing the predicted risk when spirometry was used for morbidity/mortality by an average of 1.5 +/- 1.2 percentage points (P < .001) and prolonged ventilation time by an average of 1.3 +/- 1.4 percentage points (P < .001). CONCLUSION: The use of patient history for symptoms, medication, and/or oxygen use as the only method to determine chronic lung disease for this subgroup of patients led to underreporting of chronic lung disease and underestimation of the risk for adverse outcomes. Therefore data submission to the Society of Thoracic Surgeons database should be designed to capture and correct for potential bias in the definition of chronic lung disease because the rate of spirometry in different centers in defining chronic lung disease is not regulated. PMID- 20004919 TI - Esmark elastoplasty for bleeding ventricular assist device outflow graft associated with severe coagulopathy. PMID- 20004917 TI - Outcomes after a decade of laparoscopic giant paraesophageal hernia repair. AB - OBJECTIVE: Laparoscopic repair of giant paraesophageal hernia is a complex operation requiring significant laparoscopic expertise. Our objective was to compare our current approach and outcomes for laparoscopic repair of giant paraesophageal hernia with our previous experience. METHODS: A retrospective review of patients undergoing nonemergency laparoscopic repair of giant paraesophageal hernia, stratified by early versus current era (January 1997-June 2003 and July 2003-June 2008), was performed. We evaluated clinical outcomes, barium esophagogram, and quality of life. RESULTS: Laparoscopic repair of giant paraesophageal hernia was performed in 662 patients (median age 70 years, range 19-92 years) with a median percentage of herniated stomach of 70% (range 30% 100%). With time, use of Collis gastroplasty decreased (86% to 53%), as did crural mesh reinforcement (17% to 12%). Current era patients were 50% more likely to have a Charlson comorbidity index score greater than 3. Thirty-day mortality was 1.7% (11/662). Mortality and complication rates were stable with time, despite increasing comorbid disease in current era. Postoperative gastroesophageal reflux disease health-related quality of life scores were available for 489 patients (30-month median follow-up), with good to excellent results in 90% (438/489). Radiographic recurrence (15.7%) was not associated with symptom recurrence. Reoperation occurred in 3.2% (21/662). CONCLUSIONS: With time, we have obtained significant minimally invasive experience and refined our approach to laparoscopic repair of giant paraesophageal hernia. Perioperative morbidity and mortality remain low, despite increased comorbid disease in the current era. Laparoscopic repair provided excellent patient satisfaction and symptom improvement, even with small radiographic recurrences. Reoperation rates were comparable to the best open series. PMID- 20004920 TI - Planners to the rescue: spatial planning facilitating the development of offshore wind energy. AB - The development of offshore wind energy has started to take place surprisingly quickly, especially in North European waters. This has taken the wind energy industry out of the territory of planning systems that usually govern the siting of wind farms on land, and into the world of departmental, sectoral regulation of marine activities. Although this has favoured the expansion of offshore wind energy in some respects, evidence suggests that the practice and principles of spatial planning can make an important contribution to the proper consideration of proposals for offshore wind arrays. This is especially so when a strategic planning process is put in place for marine areas, in which offshore wind is treated as part of the overall configuration of marine interests, so that adjustments can be made in the interests of wind energy. The current process of marine planning in the Netherlands is described as an illustration of this. PMID- 20004921 TI - Choline metabolism in glycinebetaine accumulating and non-accumulating near isogenic lines of Zea mays and Sorghum bicolor. AB - Glycinebetaine (GB) is a compatible solute that is accumulated by some plant species, especially under conditions leading to tissue osmotic stress. Genetic modification for accumulation of GB in an attempt to produce more stress tolerant plants has been a focus for several groups in recent years. However, attempts to increase tissue GB concentrations have been unsuccessful, with many transgenic lines accumulating far lower concentrations than naturally-occurring GB accumulators. A better understanding of the metabolic regulation of GB synthesis is necessary for successful molecular breeding and biotechnology. We utilized previously developed near-isogenic lines for GB accumulation to characterize the biochemical basis for GB deficiency in maize and sorghum. Salinity resulted in increased accumulation of choline in both accumulating and non-accumulating lines. When grown in the presence of NaCl, GB-non-accumulating lines had increased concentrations of choline and phosphocholine, but not GB. Decreased GB synthesis can be explained from the increased concentrations of phosphocholine in planta and the strong inhibition of N-phosphoethanolamine methyltransferase by phosphocholine observed in vitro. The lack of GB accumulation in GB-/- homozygous NILs was not due to the lack of the putative choline monooxygenase (the enzyme responsible for choline oxidation to betaine aldehyde) gene or protein that we describe. The previously identified bet1 locus does not appear to be choline monooxygenase. However, the lack of GB synthesis does affect the synthesis and turnover of choline moieties in GB non-accumulating lines, which may lead to alterations in overall 1-carbon metabolism in plants. PMID- 20004922 TI - In vitro and ex vivo inhibition of canine cyclooxygenase isoforms by robenacoxib: a comparative study. AB - In vitro whole blood canine assays were used to quantify the inhibitory actions of the novel non-steroidal anti-inflammatory drug (NSAID) robenacoxib on the cyclooxygenase (COX) isoenzymes, COX-1 and COX-2, in comparison with other drugs of the NSAID class. COX-1 activity was determined by measuring serum thromboxane (Tx)B(2) synthesis in blood samples allowed to clot at 37 degrees C for 1h. COX-2 activity was determined by measuring prostaglandin (PG)E(2) synthesis in blood samples incubated at 37 degrees C for 24h in the presence of lipopolysaccharide. The rank order of selectivity for inhibition of COX-2 versus COX-1 (IC(50) COX 1:IC(50) COX-2) for veterinary drugs was highest with robenacoxib (128.8) compared to deracoxib (48.5), nimesulide (29.2), S+ carprofen (17.6), meloxicam (7.3), etodolac (6.6), R- carprofen (5.8) and ketoprofen (0.88). Selectivity expressed as the clinically relevant ratio IC(20) COX-1:IC(80) COX-2 was highest for robenacoxib (19.8) compared to deracoxib (2.3), S+ carprofen (2.5), R- carprofen (2.1), nimesulide (1.8), etodolac (0.76), meloxicam (0.46) and ketoprofen (0.21). An in vivo pharmacokinetic ex vivo pharmacodynamic study in the dog established dosage and concentration-effect relationships for single oral doses of robenacoxib over the dosage range 0.5-8.0mg/kg. Values of C(max) and AUC were linearly related to dosage over the tested range. Robenacoxib did not inhibit serum TxB(2) synthesis (COX-1) ex vivo at dosages of 0.5-4.0mg/kg and produced only transient inhibition (at the 1h and 2h sampling times) at the 8mg/kg dosage. All dosages of robenacoxib (0.5-8mg/kg) produced marked, significant and dose related inhibition of PGE(2) synthesis (COX-2) ex vivo. The data demonstrate that in the dog robenacoxib is a highly selective inhibitor of the COX-2 isoform of COX, and significantly inhibits COX-2 and spares COX-1 in vivo when administered orally over the dosage range 0.5-4.0mg/kg. PMID- 20004923 TI - Peritoneal adhesion formation and reformation tracked by sequential laparoscopy: optimizing the time point for adhesiolysis. AB - BACKGROUND: In a high proportion of patients, operatively lysed adhesions reform. Using a rabbit adhesiogenesis model, this study assessed the efficacy of adhesiolysis and examined how this relates to the tissue composition of adhesions at the time of lysis. METHODS: Polypropylene meshes (5 x 3.5 cm) were implanted on the parietal peritoneum of New Zealand white rabbits. Some animals were killed 3, 7, 14, and 90 days postimplantation to obtain adhesion tissue. Adhesion formation/reformation was monitored by sequential laparoscopy in other animals kept for 90 days and in a separate experimental group subjected to adhesiolysis at 3 days postimplantation. Immune and inflammatory response markers were determined by immunohistochemical, Western blotting, and real-time reverse transcriptase polymerase chain reaction procedures in adhesion tissue; areas occupied by adhesions were quantified in meshes. RESULTS: In animals undergoing adhesiolysis, mesh areas covered by adhesions were significantly decreased at each follow-up time and affected areas became mesothelialized. Increased transforming growth factor (TGF)-beta1 expression was detected in adhesions at 3 days. Greatest TGF-beta1 and vascular endothelial growth factor (VEGF) protein expressions were observed at 7 days, whereas genetic overexpression was noted at 14 days. Active inflammatory cells peaked at the 7-day time point. CONCLUSION: Adhesions formed at 3 days; at this critical time, an adhesiolysis was effective in preventing reformation of future adhesions. TGF-beta1 gene and protein expression were increased in 3-day adhesions with respect to the omentum. Levels of active TGF-beta1 and VEGF were increased at 7 days, along with the inflammatory response at this time point related to tissue remodeling, which led to stabilization of adhesions. PMID- 20004924 TI - The impact of stress on surgical performance: a systematic review of the literature. AB - BACKGROUND: Safe surgical practice requires a combination of technical and nontechnical abilities. Both sets of skills can be impaired by intra-operative stress, compromising performance and patient safety. This systematic review aims to assess the effects of intra-operative stress on surgical performance. METHODS: A systematic search strategy was implemented to obtain relevant articles. MEDLINE, EMBASE, and PsycINFO databases were searched, and 3,547 abstracts were identified. After application of limits, 660 abstracts were retrieved for subsequent evaluation. Studies were included on the basis of predetermined inclusion criteria and independent assessment by 2 reviewers. RESULTS: In all, 22 articles formed the evidence base for this review. Key stressors included laparoscopic surgery (7 studies), bleeding (4 studies), distractions (4 studies), time pressure (3 studies), procedural complexity (3 studies), and equipment problems (2 studies). The methods for assessing stress and performance varied greatly across studies, rendering cross-study comparisons difficult. With only 7 studies assessing stress and surgical performance concurrently, establishing a direct link was challenging. Despite this shortfall, the direction of the evidence suggested that excessive stress impairs performance. Specifically, laparoscopic procedures trigger greater stress levels and poorer technical performance (3 studies), and expert surgeons experience less stress and less impaired performance compared with juniors (2 studies). Finally, 3 studies suggest that stressful crises impair surgeons' nontechnical skills (eg, communication and decision making). CONCLUSION: Surgeons are subject to many intra-operative stressors that can impair their performance. Current evidence is characterized by marked heterogeneity of research designs and variable study quality. Further research on stress and performance is required so that surgical training and clinical excellence can flourish. PMID- 20004925 TI - Porcine circovirus type 2 ORF3 protein competes with p53 in binding to Pirh2 and mediates the deregulation of p53 homeostasis. AB - The ORF3 protein of porcine circovirus type 2 (PCV2) causes apoptosis in virus infected cells and is not essential for virus replication. The ORF3 protein plays an important role in the pathogenesis of the PCV2 infection in mouse models and SPF piglets. The ORF3 protein interacts with the porcine homologue of Pirh2 (pPirh2), a p53-induced ubiquitin-protein E3 ligase, which regulates p53 ubiquitination. Here, we present our study analyzing the details of the molecular interaction between these three factors. Our experiments, in vitro and in vivo, show that ORF3 protein competes with p53 in binding to pPirh2. The amino acid residues 20 to 65 of the ORF3 protein are essential in this competitive interaction of ORF3 protein with pPirh2 over p53. The interaction of ORF3 protein with pPirh2 also leads to an alteration in the physiological cellular localization of pPirh2 and a significant reduction in the stability of pPirh2. These events contribute to the deregulation of p53 by pPirh2, leading to increased p53 levels and apoptosis of the infected cells. PMID- 20004927 TI - Novel membrane bioreactor (MBR) coupled with a nonwoven fabric filter for household wastewater treatment. AB - Conventional and modified membrane bioreactors (MBRs) are increasingly used in small-scale wastewater treatment. However, their widespread applications are hindered by their relatively high cost and operational complexity. In this study, we investigate a new concept of wastewater treatment using a nonwoven fabric filter bag (NFFB) as the membrane bioreactor. Activated sludge is charged in the nonwoven fabric filter bag and membrane filtration via the fabric is achieved under gravity flow without a suction pump. This study found that the biofilm layer formed inside the NFFB achieved 10mg/L of suspended solids in the permeate within 20 min of initial operation. The dynamic biofilter layer showed good filterability and the specific membrane resistance consisted of 0.3-1.9 x 10(12)m/kg. Due to the low F/M ratio (0.04-0.10 kg BOD(5)/m(3)/d) and the resultant low sludge yield, the reactor was operated without forming excess sludge. Although the reactor provided aerobic conditions, denitrification occurred in the biofilm layer to recover the alkalinity, thereby eliminating the need to supplement the alkalinity. This study indicates that the NFFB system provides a high potential of effective wastewater treatment with simple operation at reduced cost, and hence offer an attractive solution for widespread use in rural and sparsely populated areas. PMID- 20004926 TI - Expanding the role of 3-O sulfated heparan sulfate in herpes simplex virus type-1 entry. AB - Heparan sulfate (HS) proteoglycans are commonly exploited by multiple viruses for initial attachment to host cells. Herpes simplex virus-1 (HSV-1) is unique because it can use HS for both attachment and penetration, provided specific binding sites for HSV-1 envelope glycoprotein gD are present. The interaction with gD is mediated by specific HS moieties or 3-O sulfated HS (3-OS HS), which are generated by all but one of the seven isoforms of 3-O sulfotransferases (3 OSTs). Here we demonstrate that several common experimental cell lines express unique sets of 3-OST isoforms. While the isoforms 3-OST-3, -5 and -6 were most commonly expressed, isoforms 3-OST-2 and -4 were undetectable in the cell lines examined. Since most cell lines expressed multiple 3-OST isoforms, we addressed the significance of 3-OS HS in HSV-1 entry by down-regulating 2-O-sulfation, a prerequisite for 3-OS HS formation, by knocking down 2-OST expression by RNA interference (RNAi). 2-OST knockdown was verified by reverse-transcriptase PCR and Western blot analysis, while 3-OS HS knockdown was verified by immunofluorescence. Cells showed a significant decrease in viral entry, suggesting an important role for 3-OS HS. Implicating 3-OS HS further, cells knocked down for 2-OST expression also demonstrated decreased cell-cell fusion when cocultivated with effector cells transfected with HSV-1 glycoproteins. Our findings suggest that 3-OS HS may play an important role in HSV-1 entry into many different cell lines. PMID- 20004928 TI - Effect of shear force and solution pH on flocs breakage and re-growth formed by nano-Al(13) polymer. AB - The breakage and re-growth of flocs formed by polyaluminum chloride (PAC) and the Al(13)O(4) (OH)(24)(7+) (Al(13) for short) polymer were comparatively evaluated for the coagulation of humic acid (HA). A series of jar experiments were conducted to investigate the impacts of shear rate and solution pH on flocs breakage and re-aggregation potential. Results indicated that the responses of flocs to the increasing shear force and solution pH depend on the coagulant used. The ability of flocs to resist breakage decreased with the increasing shear rate. For all levels of shear force investigated in this study, the flocs formed by Al(13) polymer were weaker than those of PAC, whereas Al(13) polymer displayed a better recoverability than PAC. The similar results were obtained when pH of solution was changed. The flocs generated in acidic conditions were stronger and more recoverable than those generated in alkaline conditions no matter which coagulant was used. PMID- 20004929 TI - Operation of suspended-growth shortcut biological nitrogen removal (SSBNR) based on the minimum/maximum substrate concentration. AB - This study exploited the concept of the minimum/maximum substrate concentrations (MSC values) for identifying proper start-up conditions and achieving stable and low effluent total ammonium nitrogen (TAN) concentrations in suspended-growth short-cut biological nitrogen removal (SSBNR). Calculations based on the MSC concept indicated that S(Dmax), the TAN concentration above which ammonium oxidizing bacteria (AOB) are washed out, was around 450mgTAN/L at the given operating conditions of 2mg/L of dissolved oxygen and pH 8, while nitrite oxidizing bacteria (NOB) should be washed out at around 40mgTAN/L. Therefore, the experimental research was focused on the optimal TAN-concentration range for SSBNR, between 50 and 100mg/L. Experimental results showed that a nitrification reactor with initial TAN concentration above 450mg/L did not give a successful start-up. However, two days of starvation, which decreased the TAN concentration in the reactor to 95mg/L, stabilized the reaction quickly, and stable SSBNR was sustained thereafter with 80mgTAN/L and 98% nitrite accumulation in the reactor. During stable SSBNR, the removal ratio of chemical oxygen demand per nitrite nitrogen (DeltaCOD/DeltaNO(2)-N) for denitrification was 1.94gCOD/gN, which is around 55% of that required for nitrate denitrification. Based on a clone library, Nitrosomonas occupied 14% of the total cells, while the sum of Nitrobacter and Nitrospira was less than the detection cut-off of 2%, confirming the NOB were washed out during SSBNR. A spiking test that doubled the influent ammonium loading caused the TAN concentration in the reactor to reach washout for AOB, which lasted until the loading was reduced. Thus, a loading increase should be controlled carefully such that the system does not exceed the washout range for AOB. PMID- 20004930 TI - On the identity and formation routes of environmentally abundant tri- and tetrabromodibenzo-p-dioxins. AB - Ten previously unidentified polybrominated dibenzo-p-dioxins (PBDDs) were identified or tentatively identified in samples of Baltic Sea sediment and biota. Five tetrabrominated dibenzo-p-dioxins (TeBDDs) were identified using authentic reference standards, including the four most abundant (previously denoted 4U1 through 4U3). In addition, five tribromodibenzo-p-dioxins (TrBDDs) were tentatively identified using retention times and analysis of TeBDD debromination products (including a moderately abundant compound previously denoted 3U1). An attempt was made to improve the gas chromatographic separation of the major TeBDDs, but with limited success. The data generated were used as basis for a discussion on the potential PBDD formation routes. No single formation pathway could explain the PBDD patterns found. Instead, a combination of a number of potential formation processes is suggested, including enzymatic and photolytic coupling of environmentally abundant bromophenols or hydroxyl diphenyl ethers (OH PBDEs), and debromination of PBDDs to lesser brominated products. PMID- 20004931 TI - Application of thermal plasma to vitrify fly ash from municipal solid waste incinerators. AB - Several fly ash samples collected from different municipal solid waste incinerator plants were vitrified using thermal plasma furnace at 1400-1500 degrees C. After vitrification, there are obvious changes in microstructures and crystalline phases between produced slag and original ash. Toxicity characteristic leaching procedure results show that there is an excellent resistance against leaching of heavy metal ions. The residual fractions of heavy metals in slag presumably decrease in the following sequence: Cr>Ni>Cu>Zn>Pb>Cd. Almost all of PCDD/Fs could be destroyed through thermal plasma treatment. And the average decomposition efficiency of the samples vitrified by thermal plasma furnace is 100% in toxic equivalent calculations, higher than that of the samples vitrified by conventional resistance furnace. PMID- 20004932 TI - High-throughput analysis of tetracycline antibiotics and their epimers in liquid hog manure using Ultra Performance Liquid Chromatography with UV detection. AB - Antibiotics contained in animal manure can contaminate soil, groundwater and eventually surface and drinking water. To reduce the usage of antibiotics in livestock industry the EU banned their application as growth promoters in 2006. Even though the antibiotics are still used for this purpose and therefore it is necessary to control their applications. An Ultra Performance Liquid Chromatography method (UPLC) with UV detection for determination of tetracycline (TC), oxytetracycline (OTC), chlortetracycline (CTC), and doxycycline (DOX) including their epimers in the liquid hog manure was developed. The antibiotics were extracted with ethyl acetate and separated on UPLC BEH Shield RP18 column. The validated method was selective for all analytes and system suitability was assessed. Calibration curves ranged from 7.8 to 250.0mugmL(-1) with determination coefficient of 0.9999. The method limits of quantification ranged from 0.9 to 1.6mgkg(-1). Recoveries were 52.4+/-3.8%, 72.4+/-5.0%, 83.8+/-5.7% and 95.9+/ 4.7% for TC, OTC, CT, and DOX, respectively. The method was used for the determination of TC, OTC, CT, and DOX in liquid hog manure samples. PMID- 20004933 TI - Test burn with PCB-oil in a local cement kiln in Sri Lanka. AB - The production and use of polychlorinated biphenyls (PCBs) have ceased and most developed countries have disposed off their stocks long time ago. PCBs can however still be found in the environment and one important source is accumulated stocks in developing countries. Sound treatment of PCB is costly and most developing countries do not have dedicated hazardous waste incinerators or non combustion technologies available for domestic disposal and can usually not afford export. High temperature cement kilns have been used to treat organic hazardous wastes in developed countries for decades and shown to constitute a sound option if well managed and controlled. In contrast to dedicated hazardous waste incinerators and other treatment techniques, cement kilns are already in place in virtually every country and may constitute a treatment option. The objective of this study was therefore to carry out the first test burn with PCB oil in a developing country cement kiln and to assess its feasibility and destruction performance. The 3 d test burn demonstrated that the Sri Lankan cement kiln was able to destroy PCB in an irreversible and environmental sound manner without causing any new formation of PCDD/PCDF or HCB. The destruction and removal efficiency (DRE) was better than 99.9999% at the highest PCB feeding rate. PMID- 20004934 TI - Defining the borders of splenic marginal zone lymphoma: a multiparameter study. AB - Classic splenic marginal zone lymphomas are CD5-, CD10-, CD23-, CD43-, and usually IgD+ with biphasic white pulp nodules. However, the 2008 World Health Organization classification accepts splenic marginal zone lymphomas with monophasic marginal zone-like white pulp nodules and recognizes a group of unclassifiable splenic small B-cell lymphomas. To explore the relationship of classic splenic marginal zone lymphomas to these other less well-defined splenic lymphomas, a multiparameter study of 47 splenic marginal zone lymphomas and unclassifiable splenic small B-cell lymphomas was performed. Seventeen of 31 splenic marginal zone lymphomas were biphasic, and 14 were monophasic (90%-100% marginal zone-like white pulp nodules). Sixteen cases were unclassifiable splenic small B-cell lymphomas, most lacking a marginal zone-type component. There were many clinical similarities between the 3 groups, including similar survivals. Monophasic and unclassifiable cases were less likely to have a typical splenic marginal zone lymphoma phenotype (28.6%, 23.1%) compared with biphasic cases (86.7%), usually because of IgD negativity (P < .003). Thirty-four of 42 (81%) cases had cytogenetic abnormalities by fluorescence in situ hybridization; and 17 of 20 (85%), by classical cytogenetics. The most frequent fluorescence in situ hybridization abnormalities among the splenic marginal zone lymphomas were del(7)(q31) (26%), +12 (25%), and +3q27 (27%); and among the unclassifiable cases, +12 (50%) and +3q27 (36%). Five of 6 unclassifiable cases with exclusively small non-marginal zone-like lymphocytes involving both white and red pulp had +12 compared with 9 of 34 other cases (P < .02). CDK6 (2 cases) and BCL3 (1 case) rearrangements were only seen in the unclassifiable group. These results support including both biphasic and monophasic cases as splenic marginal zone lymphomas, but suggest that the lack of a non-marginal zone-like population in the monophasic group is associated with some biologic differences. They also demonstrate a relatively large proportion of unclassifiable cases, including a group with frequent +12. PMID- 20004935 TI - Pleomorphic and dedifferentiated leiomyosarcoma: clinicopathologic and immunohistochemical study of 41 cases. AB - In this article, we supplement the few published articles by describing the clinical and pathologic features of pleomorphic and dedifferentiated leiomyosarcoma from 41 patients (27 women and 14 men) with an age range of 25 to 75 years (mean, 56.5 years), representing the largest cohort reported to date. The typical leiomyosarcoma component accounted for <5% to 60% (mean, 15%) of the tumor. The pleomorphic sarcoma component was composed of polygonal cells in 57% of cases, spindle cells in 21%, a combination of polygonal, epithelioid, rhabdoid, and/or spindle cells in 18%, and predominantly epithelioid cells in 3%. The classical leiomyosarcoma component was positive for at least one myogenic immunohistochemical marker in 29 tumors tested; smooth muscle actin in 100% (27/27), calponin in 90% (9/10), muscle-specific actin in 90% (10/11), desmin in 86% (23/27), smooth muscle myosin heavy chain (SMMS-1) in 67% (4/6), and caldesmon in 57% (4/7). The pleomorphic sarcoma component was reactive for at least one muscle marker in 77% (23/30) of cases; smooth muscle actin in 63% (17/27), calponin in 60% (6/10), SMMS-1 in 60% (3/5), desmin in 59% (16/27), muscle-specific actin in 40% (4/10), and caldesmon in 29% (2/7). The classical leiomyosarcoma component was often strongly positive for myogenic markers, and the pleomorphic sarcoma component usually showed focal and less intense immunoreactivity. Based on staining for muscle markers in the pleomorphic component, twenty-three cases were designated as pleomorphic leiomyosarcoma, and 7 cases were designated as dedifferentiated leiomyosarcoma (negative for all muscle markers used). Eleven cases, in which tissue was not available for immunhistochemical stains, the question of pleomorphic versus dedifferentiated leiomyosarcoma could not be answered. The incidence of metastasis was 89% (32/36) and the mortality rate was 50% (18/36) at last follow-up (3-104 months; mean, 27.5 months). PMID- 20004936 TI - Any proportion of ductal adenocarcinoma in radical prostatectomy specimens predicts extraprostatic extension. AB - Ductal adenocarcinoma of the prostate is an aggressive malignancy, often presenting at an advanced stage. In mixed ductal and acinar adenocarcinomas, the relationship between the proportion of the ductal component of the tumor and the pathologic stage and whether or not aggressive behavior is simply a function of grade remains undetermined. From 268 consecutive radical prostatectomies undertaken as a curative procedure for clinical localized prostate cancer, we identified 34 cases (12.7%) with ductal adenocarcinoma of the prostate comprising 5% to 100% of the total tumor volume. For cases with a ductal adenocarcinoma of the prostate component, the mean age at diagnosis of 60 years (range 49-69 years), mean serum prostate-specific antigen of 8.4 ng/mL (range, 0.8-21 ng/mL) and positive surgical margin rate of 17.6% did not differ significantly from that of the pure adenocarcinoma group. All 34 patients with ductal adenocarcinoma of the prostate had peripheral zone involvement while 16 (46%) also had transition zone involvement. Twenty-five (73%) cases with ductal adenocarcinoma of the prostate had extraprostatic extension (pT3), which compared to 32.9% with acinar adenocarcinoma. The presence of ductal adenocarcinoma of the prostate (P < .0001), high tumor volume (P = .001) and Gleason score >7 (P = .04) significantly predicted pT3 staging category, and the presence of ductal adenocarcinoma of the prostate remained a significant predictor for pT3, after adjusting for tumor volume and Gleason score >7. The proportion of ductal adenocarcinoma of the prostate did not significantly modify the strength of the observed association with pathological stage. In view of the significant association with extraprostatic extension we would recommend that in both core biopsies and radical prostatectomy specimens any proportion of ductal adenocarcinoma of the prostate should be reported. PMID- 20004937 TI - Post-mortem pathologic and genetic studies in "dead in bed syndrome" cases in type 1 diabetes mellitus. AB - Dead in bed syndrome is a poorly understood cause of sudden death in young people with type 1 diabetes. The underlying cause remains unknown. One possible explanation may involve prolongation of the QT interval followed by a terminal malignant arrhythmia. Risk factors associated with QT interval prolongation include hypoglycemia and cardiac autonomic neuropathy. We sought to identify myocardial cellular changes and genetic influences that may contribute to the pathogenesis of dead in bed syndrome. Post-mortem reports between 1994 and 2006 from the 2 largest Departments of Forensic Medicine in Australia were reviewed for dead in bed syndrome cases. Post-mortem heart sections were immunohistochemically stained for collagen types I and III and connective tissue growth factor (CTGF). Genomic DNA was prepared from post-mortem samples, and genetic analysis was performed in the SCN5A, G6PC, PHOX2B, and CTGF genes. Twenty two dead in bed syndrome cases were identified and staining of heart sections for collagen I and III, and CTGF showed no differences between dead in bed syndrome cases and controls. Genetic screening of SCN5A revealed 3 silent polymorphisms A29A, E1061E, and D1819D and 1 protein-changing variant H558R. No genetic variants were found in G6PC, PHOX2B, and CTGF, and dead in bed syndrome cases were not associated with the G-945C CTGF promoter polymorphism. In conclusion, this study is the first to investigate potential pathogenic mechanisms underlying the dead in bed syndrome in type 1 diabetes with the results substantially adding to knowledge of this condition. Understanding the causes and triggers of dead in bed syndrome will be critical in facilitating the identification of patients with type 1 diabetes at highest risk of developing sudden death. PMID- 20004938 TI - Flat epithelial atypia is a common subtype of B3 breast lesions and is associated with noninvasive cancer but not with invasive cancer in final excision histology. AB - The biological behavior and the optimal management of benign breast lesions with uncertain malignant potential, the so-called B3 lesions, found in breast needle core biopsies is still under debate. We addressed this study to compare histologic findings in B3 needle core biopsies with final excision specimens to determine associated rates of malignancy. Consecutive needle core biopsies were performed in a 3-year period (January 1, 2006-December 31, 2008). Biopsies were image-guided (31 by ultrasound, 85 stereotactic vacuum-assisted, 6 unknown) for evaluation of breast abnormalities. We reviewed 122 needle core biopsies with B3 lesions of 91 symptomatic patients and 31 screen-detected women and compared the B3 histologic subtypes with the final excision histology. A total of 1845 needle core biopsies were performed and B3 lesions comprised 6.6% of all B categories. The most common histologic subtype in biopsies was flat epithelia atypia in 35.2%, followed by papillary lesions in 21% and atypical ductal hyperplasia in 20%. Reports on excision specimens were available in 66% (81 patients). Final excision histology was benign in 73 (90.2%) and malignant in 8 (9.8%) patients (2 invasive cancer, 6 ductal carcinoma in situ). Of all B3 subtypes, atypical ductal hyperplasia and flat epithelial atypia were associated with malignancy, whereas only atypical ductal hyperplasia was accompanied by invasive cancer. Of all lesions, flat epithelial atypia was most frequently found in excision specimens (18%). In our study, flat epithelial atypia and atypical ductal hyperplasia are common lesions of the B3 category in needle core biopsies of the breast. Both lesions are associated with malignancy, whereas only atypical ductal hyperplasia was related to invasive cancer. We conclude that an excision biopsy after diagnosis of flat epithelial atypia is recommended depending on clinical and radiologic findings. PMID- 20004940 TI - Large cell calcifying Sertoli cell tumor: a clinicopathologic study of 1 malignant and 3 benign tumors using histomorphology, immunohistochemistry, ultrastructure, comparative genomic hybridization, and polymerase chain reaction analysis of the PRKAR1A gene. AB - Four cases of large cell calcifying Sertoli cell tumor, 3 benign and 1 malignant, with no clinical signs of Carney complex or Peutz-Jeghers syndrome are reported with results of histologic, immunohistochemical, ultrastructural, and comparative genomic hybridization studies. Analysis of PRKAR1A gene was performed on 2 cases. The age range of the patients was 19 to 54 years. The patient with a malignant large cell calcifying Sertoli cell tumor died of disease 4 years after surgery. Patients with benign tumors have had an uneventful follow-up for 1 and 3 years. All tumors were well circumscribed, unencapsulated, and composed of solid sheets, irregular cords, tubular structures, and nests in a fibrous and/or myxoid stroma with cellular atypia in the malignant case. All tumors showed diffuse immunoreactivity for inhibin, vimentin, calretinin, and S100 protein. Focal positivity for cytokeratin (AE1/AE3) was noticed in 1 case. Tumors were negative for CAM 5.2, Mic-2, Melan-A laminin, placental alkaline phosphatase, and alpha fetoprotein. The proliferation index was 5% and 10% for 2 of the benign tumors and 30% for the malignant tumor. Comparative genomic hybridization was performed in 2 cases. There was no evidence of any major chromosomal changes. In one case, no PRKAR1A gene mutation was found. In the other case, a heterozygous shift mutation c.65_84dup was found, despite the absence of other clinical signs of Carney complex or Peutz-Jeghers syndrome. Although the combination of large cell calcifying Sertoli cell tumor and PRKAR1A mutation fulfills the criteria for establishing a diagnosis of Carney complex, the clinical relevance of finding a PRKAR1A gene mutation in a patient without any clinical signs of Carney complex or Peutz-Jeghers syndrome remains to be established. PMID- 20004941 TI - Correlation of overexpression of HMGA1 and HMGA2 with poor tumor differentiation, invasion, and proliferation associated with let-7 down-regulation in retinoblastomas. AB - In addition to RB1, the causative genes involved in the tumorigenesis and progression of retinoblastomas remain to be elucidated. High-mobility group A1 and high-mobility group A2 proteins are expressed at high levels in various benign and malignant tumors and are associated with expressions of malignant phenotypes and poor prognoses. Reduction in let-7 expression levels was detected in cancers; it may be related to high-mobility group A1 and high-mobility group A2 overexpressions. Little is known about the correlations among high-mobility group A1, high-mobility group A2, and let-7 expression and clinicopathologic features of retinoblastoma. In our study, the expressions of high-mobility group A1 and high-mobility group A2 were studied in 44 retinoblastomas by immunohistochemical analysis. Semiquantitative reverse transcription-polymerase chain reaction was used to assay the let-7 expression levels in 28 nontumor retina and 44 tumor samples. Nuclear immunostaining of high-mobility group A1 and high-mobility group A2 was frequently observed in retinoblastomas (68% and 75%, respectively). Expression levels of both high-mobility group A1 and high-mobility group A2 were significantly higher in poorly differentiated retinoblastomas than in well-differentiated retinoblastomas (P < .05 and P < .0001, respectively). In addition, overexpressions of high-mobility group A1 and high-mobility group A2 were more frequently detected in poorly differentiated tumors than in well differentiated tumors (P < .01 and P = .0001, respectively). High-mobility group A2 expression levels were significantly higher in invasive tumors than in noninvasive tumors (P < .05). In addition, the MIB-1 labeling index was higher in poorly differentiated tumors than in well-differentiated tumors (P < .0001). Our study revealed that high-mobility group A1 and high-mobility group A2 expressions correlated with the MIB-1 labeling index (R = 0.327, P = .029; R = 0.602, P < .0001; respectively). The let-7 was expressed in high levels in all 28 nontumor retina samples. However, reduced expression levels of let-7 were observed in 17 (39%) tumors. A potentially inverse correlation exists between the expression levels of let-7 and high-mobility group A1 (r = -0.247, P = .105). In addition, a significantly inverse association was detected between let-7 and high-mobility group A2 and MIB-1 labeling index (r = -0.31, P = .04; r = -0.392, P = .007, respectively). Our findings imply that the overexpressions of high-mobility group A1, high-mobility group A2, and down-regulation of let-7 may be associated with tumorigenesis and progression of retinoblastomas. PMID- 20004942 TI - Characterization of enteroglial cells and denervation process in chagasic patients with and without megaesophagus. AB - Chagas disease is caused by infestation with the parasite Trypanosoma cruzi, and some patients who are serologically positive develop chronic megaesophagus, whereas others are symptom-free. Gastrointestinal form of Chagas disease involves an inflammatory invasion of the enteric plexuses and degeneration of enteric neurons and previous works related that enteroglial cells would be involved in enteric inflammatory responses. Because of this, the aims of this study were to determine the relation of enteroglial cells with the denervation process in chagasic patients with and without megaesophagus and seronegative individuals. Our results indicated that the innervation of the esophageal muscle was substantially reduced in patients with megaesophagus, but asymptomatic seropositive subjects were not different to seronegative controls. Besides, patients with megaesophagus had significant decreased of enteroglial cells labeled with S-100 and glial fibrillary acidic protein, whereas patients without megaesophagus presented an increased of both labels. We believe that enteroglial cells would operate a mechanism of defense in the enteric nervous system against the Trypanosoma cruzi infection, which could prevent the organ denervation and preserve the esophagus function. PMID- 20004943 TI - Quantitative expression of VEGF, VEGF-R1, VEGF-R2, and VEGF-R3 in melanoma tissue microarrays. AB - Angiogenesis is required for progression and metastasis of melanoma. Analysis of angiogenic molecules in benign and malignant tissues may allow identification of markers useful for prediction of sensitivity to antiangiogenic agents. We hypothesized that differential expression of vascular endothelial growth factor (VEGF) and its receptors VEGF-R1, VEGF-R2, and VEGF-R3 would be higher in melanomas than nevi and higher in advanced melanoma. Using automated quantitative analysis, we quantified VEGF, -R1, -R2 and -R3 expression in melanoma tissue microarrays composed of 540 nevi and 468 melanoma specimens (198 primaries, 270 metastases). VEGF, VEGF-R1, VEGF-R2, and VEGF-R3 expression was significantly higher in melanomas than nevi by unpaired t tests (P < .0001). VEGF-R2 expression was higher in metastatic specimens (P < .0001), but VEGF-R3 expression was higher in primaries (P < .0001). VEGF was coexpressed with all 3 receptors when assessed by Spearman's rank correlation. VEGF, VEGF-R1, VEGF-R2, and VEGF-R3 expression is higher in melanomas than nevi. Higher expression of VEGF-R2 was found in metastases versus primaries, supporting the idea that selection for an angiogenic phenotype in metastatic melanoma is conferred via up-regulation of VEGF-R2. However, higher expression of VEGF-R3 was seen on primary lesions, potentially implicating this receptor in initiation of lymphatic tumor spread. Clinical trials using antiangiogenic agents in melanoma should include correlative assays of VEGF, VEGF-R1, VEGF-R2, and VEGF-R3 as biomarkers of response to therapy, preferably using quantitative methods such as automated quantitative analysis. Such assessments could assist with evaluation of these molecules as therapeutic targets in melanoma, ultimately facilitating improved selection of patients for treatment. PMID- 20004944 TI - Lymphatic and blood vessels in scleroderma skin, a morphometric analysis. AB - Vascular involvement is frequent in systemic sclerosis, but the role of the lymphatic vasculature is poorly known. Our aim was to evaluate lymphatic vessels in systemic sclerosis skin lesions. We studied skin forearm biopsies of 9 patients with systemic sclerosis and 7 age-matched controls. Lymphatic vessels were labeled with the monoclonal antibody D2-40 and blood vessels with a polyclonal antibody to von Willebrand Factor. All blood and lymphatic vessels present in each section were counted and total area, inner luminal area, and shape factors were measured. The number of blood and lymphatic vessels in papillary dermis was greater and their diameter lower than in reticular dermis both in systemic sclerosis and controls. In the reticular dermis, the number of lymphatic vessels was markedly reduced in systemic sclerosis (4.9 +/- 1.1 microm( 2) versus 8.9 +/- 1.2 microm(-2)P = .03), and a similar trend was observed in papillary dermis (8.4 +/- 3.7 microm(-2) versus 8.1 +/- 5.3 microm(-2)). Interestingly, the number of periglandular lymphatics in systemic sclerosis was not different from controls. The inner luminal area (possibly indicating compensatory dilation) of lymphatic vessels, particularly the periglandular ones, was greater in systemic sclerosis than in controls. No differences were observed in the number of blood vessels, but the percentage of blood vessel profiles (without lumen) was significantly less in systemic sclerosis both in papillary and in reticular dermis. In conclusion, our data show that skin lesions in systemic sclerosis are characterized by a selective rarefaction of lymphatic vasculature that spares periglandular vessels and that might have a pathogenic role in the evolution and in the clinical manifestations of the disease. PMID- 20004945 TI - Case report: focal nesidioblastosis ("nesidioblastoma") in an adult. AB - Adult nesidioblastosis is an uncommon cause of hyperinsulinemic hypoglycemia characterized by diffuse islet hyperplasia with beta-cell hypertrophy and atypia. The cause of nesidioblastosis in adults is unclear but may be different from nesidioblastosis in infants. In contrast to infants, a focal form of adult nesidioblastosis (ie, "nesidioblastoma") has not been documented, although proposed. We report a 44-year-old man with symptomatic hypoglycemia and localized nesidioblastosis treated with surgical enucleation resulting in normalization of blood glucose. Postoperative euglycemia has persisted in this patient to date (4 months at the time of manuscript submission). PMID- 20004946 TI - Beclin 1 and LC3 autophagic gene expression in cutaneous melanocytic lesions. AB - Beclin 1 and LC3 autophagic genes are altered in several human cancer types. This study was designed to assess the expression of Beclin 1 and LC3 in cutaneous melanocytic lesions, in which they have not yet been investigated. In melanoma, we correlated their expression with conventional histopathologic prognostic factors. In 149 lesions, including benign nevi, dysplastic nevi, radial growth phase melanomas, vertical growth phase melanomas, and melanoma metastases, proteins were evaluated by immunohistochemistry, and, in representative cases of benign nevi, vertical growth phase melanomas and melanoma metastases were evaluated by Western blotting. In most lesions, messenger RNA level was also assessed by real-time reverse transcriptase polymerase chain reaction. Both genes were expressed in all the investigated conditions. Beclin 1 cytoplasmic protein and messenger RNA, as well as LC3 messenger RNA, significantly decreased with tumor progression (P < .05). The percentage of cases with high cytoplasmic expression of beclin 1 from 100% in benign nevi declined to 86.4% in dysplastic nevi, 54.5% in radial growth phase melanomas, 54.3% in vertical growth phase melanomas, and 26.7% in melanoma metastases. The lowest expression of LC3 II protein was observed in melanoma metastases (53.3% of cases) (P < .05); LC3 II protein overexpression was, however, found in several nonbenign lesions, with the highest percentage (45.5%) in radial growth phase melanomas. LC3 II protein expression was inversely correlated to thickness, ulceration, and mitotic rate. In a multivariate analysis, messenger RNAs for both genes discriminated between nonmalignant (benign and dysplastic nevi) and malignant (radial, vertical growth phase melanomas, and melanoma metastases) lesions. Our results, therefore, indicate that beclin 1 and LC3 II autophagic gene expression is altered also in melanocytic neoplasms. PMID- 20004947 TI - Reduction of CD44(+)/CD24(-) breast cancer cells by conventional cytotoxic chemotherapy. AB - Breast cancer cells with the CD44(+)/CD24(-) phenotype have been associated with stem cell properties. To analyze effects of cytotoxic chemotherapy on these cells, we examined a series of 50 breast carcinomas before and after neoadjuvant chemotherapy with epirubicin/cyclophosphamide using double immunofluorescence. Before treatment, an average of 4.4% of the tumor cells displayed a CD44(+)/CD24( ) phenotype. However, after chemotherapy, the frequency of CD44(+)/CD24(-) cells dropped to 2% (P = .008). To test this unexpected finding, we analyzed a second collective of 16 patients that preoperatively had received either 4 cycles of doxorubicin/pemetrexed, followed by 4 cycles of docetaxel or 4 cycles of doxorubicin/cyclophosphamide, followed by 4 cycles of docetaxel with similar results (8.7% CD44(+)/CD24(-) cells on average before and 1.1% after chemotherapy). In addition, no association was observed between the frequency of CD44(+)/CD24(-) cells and the response to chemotherapy or patient survival. However, patients with tumors containing high numbers of CD44(+)/CD24(-) cells more frequently developed bone metastases in the course of disease. In conclusion, our findings challenge the proposed role of CD44(+)/CD24(-) cells as cancer stem cells in tumor resistance to chemotherapy as they apparently are not selected by conventional cytotoxic agents. PMID- 20004948 TI - IMP3 expression is correlated with histologic grade of lung adenocarcinoma. AB - Insulin-like growth factor II mRNA binding protein 3 is an oncofetal protein that is expressed in multiple malignancies. This study aimed to determine the correlation of insulin-like growth factor II mRNA binding protein 3 expression with histologic grade of lung adenocarcinoma. Eighty-nine cases, including 11 atypical adenomatous hyperplasias, 10 pure bronchioloalveolar carcinomas, 36 well differentiated adenocarcinomas and 41 moderately or poorly differentiated adenocarcinomas, were immunohistochemically studied using a monoclonal antibody against insulin-like growth factor II mRNA binding protein 3. Twenty-nine (70.7%) of 41 moderately to poorly differentiated adenocarcinomas were positive for insulin-like growth factor II mRNA binding protein 3, with 26 (89.7%) tumors demonstrating either a strong staining or staining in greater than 30% of tumor cells. Four (40.0%) of 10 bronchioloalveolar carcinomas and 13 (36.1%) of 36 well differentiated adenocarcinomas exhibited insulin-like growth factor II mRNA binding protein 3 positivity with a variable degree and percentage of tumor cells staining. When bronchioloalveolar carcinomas were present in a pure form or as a component of adenocarcinomas, positive insulin-like growth factor II mRNA binding protein 3 staining was always patchy, with less than 20% of tumor cells stained. Overall, the frequency of positive insulin-like growth factor II mRNA binding protein 3 staining was lower in bronchioloalveolar carcinomas and well differentiated adenocarcinomas compared to moderately/poorly differentiated adenocarcinomas (P < .01). No insulin-like growth factor II mRNA binding protein 3 signal was detected in any case of atypical adenomatous hyperplasia. These findings show that insulin-like growth factor II mRNA binding protein 3 is strongly and diffusely expressed in a large proportion of moderately/poorly differentiated lung adenocarcinomas, in particular in the solid component of mixed subtype adenocarcinomas, less frequently expressed in well-differentiated adenocarcinomas and bronchioloalveolar carcinomas, and negative in atypical adenomatous hyperplasias. The higher frequency of expression in moderately/poorly differentiated adenocarcinomas suggests that insulin-like growth factor II mRNA binding protein 3 expression may be associated with an aggressive biological behavior. PMID- 20004949 TI - Interstitial inflammation in Alport syndrome. AB - The Alport syndrome is a hereditary glomerular disease linked to structural abnormalities of collagen IV. In a mouse model of Alport syndrome, the interstitial lymphocyte influx was important for disease progression. CXCR3 is a chemokine receptor involved in lymphocyte recruitment to the kidney. We hypothesized that CXCR3-positive T cells might be involved in human Alport syndrome. Immunohistochemistry was performed on formalin-fixed, paraffin-embedded biopsies from 17 patients with Alport syndrome, 10 with immunoglobulin A (IgA) nephropathy, and 11 healthy donor kidneys. We investigated the expression of the alpha5 chain of collagen IV to confirm the morphologic diagnosis, the chemokine receptor CXCR3 and CD3-positive T cells. Alport syndrome biopsies demonstrated a complete loss of the alpha5 chain of collagen IV from the glomerular basement membrane and the morphologic features consistent with Alport syndrome on electron microscopy. A prominent number of CXCR3-positive cells were found in the tubulointerstitium. Most of the CXCR3-positive cells were CD3-positive T cells, demonstrated by double-labeling in selected biopsies. The number of CXCR3 positive cells in kidneys with Alport syndrome correlated with serum creatinine (P < .05) and with morphologic features of a progressive disease (eg, interstitial fibrosis, glomerulosclerosis, and tubular atrophy). The severity of interstitial CXCR3-positive cell influx was similar in Alport syndrome as compared to immunoglobulin A nephropathy. The noninflammatory glomerular lesion of Alport syndrome is associated with prominent interstitial accumulation of CD3- and CXCR3-positive lymphocytes. The degree of infiltration correlated with renal function. We speculate that targeting T lymphocytes, for example, by CXCR3 blocking agents, might be a novel approach to inhibit disease progression in patients with Alport syndrome. PMID- 20004950 TI - Expression of p16 in benign and malignant cystic squamous lesions of the neck. AB - Metastatic cystic squamous cell carcinomas of the neck often harbor human papillomavirus 16 and, in turn, overexpress p16. P16 immunohistochemistry could be useful in the evaluation of patients who present with cystic squamous lesions of the neck, particularly when the distinction between a benign lymphoepithelial cyst and a metastatic squamous cell carcinoma cannot be easily resolved on clinical or pathologic grounds. Implementation of this strategy, however, awaits a description of p16 expression in benign lymphoepithelial cysts. The purpose of this study was to evaluate p16 staining in cystic squamous lesions of the neck with an emphasis on benign lymphoepithelial cysts. P16 immunohistochemistry was performed on tissue sections and fine needle aspirates of benign (n = 49) and malignant (n = 16) squamous lesions of the neck. P16-positive cases were further evaluated by human papillomavirus 16 in situ hybridization. P16 staining was seen in the tissue sections of 16 of 37 (43%) benign lymphoepithelial cysts. P16 staining tended to localize to regions of the squamous epithelium penetrated by interdigitating lymphocytes. In the aspirates, p16 staining was noted in 5 of 12 (42%) benign lymphoepithelial cysts and in 3 of 16 (19%) cystic squamous cell carcinomas. Human papillomavirus 16 was detected in the 3 p16-positive cystic squamous cell carcinomas but in none of the p16-positive benign lymphoepithelial cysts. P16 overexpression is not always linked to high-risk human papillomavirus integration, but may be intrinsic to the reticulated epithelium that lines benign lymphoepithelial cysts. This observation limits the role of p16 staining as a surrogate marker of human papillomavirus 16 infection and as a diagnostic tool in separating benign from malignant cystic squamous lesions of the neck. PMID- 20004951 TI - If not, why not? Reasons why Canadian postgraduate trainees chose--or did not choose--to become pathologists. AB - Pathology has been frequently identified in the literature as an unpopular choice for medical students. For many years, there have been predictions that this unpopularity would lead to inadequate pathologist numbers, which would in turn contribute to poor quality patient care. In Canada, the predicted crisis has become a reality: after a high-profile failure of laboratory quality, a public inquiry reported that poor pathology recruitment was partially responsible and recommended that medical schools take steps to make pathology more attractive to medical students. There are several published studies into pathology recruitment, but none has asked nonpathology residents why they did not choose pathology. This study uses qualitative techniques to investigate why pathology residents chose to specialize in pathology and why clinical residents rejected a pathology career. Pathology residents across Canada were surveyed, as were clinical (nonpathology) residents in every residency training program at the University of British Columbia in Vancouver, Canada. Pathology residents overwhelmingly cited various attractive features of pathology practice, including its academic nature, the opportunity to explore basic pathogenesis, and its interesting and varied daily work. Most clinical residents rejected pathology because they preferred direct patient contact; however, a sizable minority blamed insufficient or inadequate medical school experiences in pathology. Clinical residents also cited several misconceptions and stereotypes about pathology, including misunderstandings about the role of pathologists and the nature of pathology practice. The reasons why clinical residents rejected pathology careers may provide guidance in improving pathology recruitment of medical students. PMID- 20004952 TI - Thyroid transcription factor 1 expression in ovarian carcinomas is an independent prognostic factor. AB - Tthyroid transcription factor 1 is a marker of lung and thyroid carcinomas, but thyroid transcription factor 1 immunoreactivity is seen in other malignancies. We examined the incidence of thyroid transcription factor 1 expression in gynecologic tumors in Japanese patients, and we further evaluated the presence of epidermal growth factor receptor mutations in thyroid transcription factor 1 positive gynecologic malignancies. A total of 186 patient samples collected at our hospitals between 1991 and 2006 were analyzed, and these specimens consisted of 83 ovarian carcinomas, 55 endometrioid endometrial adenocarcinomas of the uterus, 28 cervical adenocarcinomas of the uterus, and 20 leiomyosarcomas of the uterus. Thyroid transcription factor 1 expression was assessed by immunohistochemistry. The presence of epidermal growth factor receptor mutations was investigated by polymerase chain reaction analyses. Thyroid transcription factor 1 was detected in the nuclei of 11 ovarian carcinomas (13%) and 5 endometrioid adenocarcinomas (10%) of the uterus. In patients with ovarian carcinoma, thyroid transcription factor 1 staining was associated with significantly improved progression-free (P = .017) and overall survival (P = .017) using univariate analysis. Multivariate analysis identified thyroid transcription factor 1 expression as an independent prognostic factor for ovarian cancer (P = .0467). No epidermal growth factor receptor mutations were found in our study. Thyroid transcription factor 1 is expressed with relatively low frequencies in gynecologic malignancies, but thyroid transcription factor 1 expression confers a better prognosis in patients with ovarian cancer. No epidermal growth factor receptor mutations were found in the thyroid transcription factor 1-positive gynecologic malignancies, and we were unable to establish a relationship between epidermal growth factor receptor mutations and thyroid transcription factor 1 immunopositivity, as was previously shown for lung cancer. PMID- 20004953 TI - Clinically occult interstitial fibrosis in smokers: classification and significance of a surprisingly common finding in lobectomy specimens. AB - This study reports the presence of surprisingly frequent and often severe interstitial fibrosis in cigarette smokers with no clinical evidence of interstitial lung disease. Twenty-three lobectomy specimens excised for neoplasms, including 20 from smokers, were extensively sampled, and examined semi quantitatively for interstitial fibrosis, fibroblast foci, peribronchiolar metaplasia, honey-comb change, emphysema, and respiratory bronchiolitis. Interstitial fibrosis involving greater than 25% of slides was identified in 12 of 20 smokers (60%), but in none of the three never-smokers. Three cases were classified as specific forms of interstitial lung disease, including one each of usual interstitial pneumonia, Langerhans cell histiocytosis, and asbestosis. The remaining 9 cases did not fit with a named interstitial lung disease and were considered to represent examples of smoking-related interstitial fibrosis. This lesion was characterized by varying degrees of alveolar septal widening by collagen deposition along with emphysema and respiratory bronchiolitis. The fibrosis occurred both in subpleural and in deeper parenchyma. It surrounded enlarged airspaces of emphysema, but it also involved non-emphysematous parenchyma. Clinical progression was not documented in any case, although follow up was short. These observations highlight the spectrum of unexpected fibrosis that is frequently encountered in lobectomy specimens from cigarette smokers. Additional investigation will be required to determine the clinical significance of smoking-related interstitial fibrosis and its relationship, if any, to other smoking-related diseases. It is important, however, that smoking-related interstitial fibrosis be distinguished from specific forms of fibrosing lung disease that may be associated with poor prognoses, especially usual interstitial pneumonia. PMID- 20004954 TI - Is there a role for B-cell depletion as therapy for scleroderma? A case report and review of the literature. AB - OBJECTIVES: Rituximab (RTX) has been successfully used in the treatment of several rheumatic diseases with an acceptable safety profile. We present herein a patient with systemic sclerosis (SSc) who exhibited significant improvement of his lung function and skin fibrosis following RTX administration, and review the literature regarding the role of B-cells in SSc and the potential efficacy of RTX in its treatment. METHODS: We performed an internet search using the keywords systemic sclerosis, scleroderma, rituximab, B-cells, fibrosis, interstitial lung disease (ILD), and therapy. RESULTS: Our patient, a 40-year old man with severe SSc-associated ILD, received 4 courses of RTX. The patient's lung function improved; forced vital capacity and diffusing capacity of carbon monoxide reached values of 35% and 33%, respectively, compared with 30% and 14% of pretreatment values. Skin thickening assessed clinically and histologically improved as well. Several lines of evidence suggest that B-cells may have a pathogenic role in SSc. B-cells from tight skin mice--an animal model of SSc--exhibit chronic hyperactivity; likewise, B-cells from patients with SSc overexpress CD19 and are chronically activated. Furthermore, studies have revealed that B-cell genes were specifically transcribed in SSc skin and that B-cell infiltration was a prominent feature of SSc-associated ILD. The potential clinical efficacy of RTX in SSc has been explored in a limited number of patients with encouraging results. Preliminary data suggest that RTX may favorably affect skin as well as lung disease in SSc. CONCLUSIONS: Several basic research data underscore the potential pathogenic role of B-cells in SSc and clinical evidence suggests that RTX might be a therapeutic option in SSc. Large-scale multicenter studies are needed to evaluate the potential clinical efficacy of RTX in SSc. PMID- 20004955 TI - Anti-factor IXa Aptamer reduces propagation of thrombin generation in plasma anticoagulated with warfarin. AB - BACKGROUND: Warfarin is routinely used in the prevention and treatment of prothrombotic events. During initiation of warfarin therapy levels of factor (F) VII and protein C decrease rapidly but prothrombin, FIX and FX decline much slower. Therefore, propagation of thrombin generation (TG) remains unaffected much longer, increasing the risk of inadequate anticoagulation. Recently, a novel agent, anti-IXa aptamer, RB006, has been developed. Therefore, we have evaluated the in vitro effects of this agent in warfarin plasma. METHODS: The investigation consisted of two parts. First, a computer simulated time course of TG with warfarin alone and in combination with FIXa inhibition was evaluated and, second, normal volunteer, protein C deficient, FVII deficient and commercial warfarin plasmas (INR 2.1 and 3.1) were spiked with increasing concentrations of aptamer (0-24 microg/ml) and its anticoagulant effects were evaluated using prothrombin time (PT), activated partial thromboplastin time (aPTT) and TG with tissue factor and Actin as activators. Direct effects of aptamer on protein C were also evaluated. RESULTS: Simulation of coagulation during warfarin induction showed that TG can be significantly delayed and decreased by inhibiting FIXa (i.e., with anti-FIXa aptamer). The anti-FIXa aptamer inhibited TG in all tested plasmas, but was most efficacious in warfarin and FVII deficient plasma. The aptamer itself did not inhibit protein C and had no effect on PT, but concentration-dependently increased aPTT. CONCLUSION: The anti-FIXa aptamer potentiates the inhibitory effects of warfarin on TG, and may fill the need as an adjuvant agent during initiation of warfarin therapy. PMID- 20004956 TI - Incidence and management of bevacizumab-associated gastrointestinal perforations in patients with recurrent ovarian carcinoma. AB - OBJECTIVE: The objective of this study was to examine the incidence and management of bevacizumab-associated gastrointestinal (GI) perforations in patients with recurrent ovarian carcinoma. METHODS: We identified all patients who received bevacizumab off protocol from August 2004-August 2008. We examined their medical records for reports of confirmed GI perforation, associated clinicopathological factors, treatment, and outcomes. RESULTS: Six (4%) of 160 patients with ovarian carcinoma who had been treated with bevacizumab developed GI perforations, with a median of 4 (range, 2-8) previous cytotoxic regimens. The median serum CA-125 at the start of treatment was 228 U/mL (range, 50-3106 U/mL). The median number of bevacizumab cycles prior to perforation was 10.5 (range, 2 20). The median time from the last bevacizumab dose to diagnosis of GI perforation was 13 days (range, 1-28 days). Four (67%) patients underwent an exploratory surgery. At laparotomy, one had a gastric perforation and one had an appendiceal perforation; the site of perforation could not be identified in the other 2 Two patients (33%) were managed conservatively-one with a PEG tube and the other with supportive care. The median time of death from the date of diagnosis of GI perforation was 27 days (range, 4-326 days). Only two patients one with a gastric and the other with an appendiceal perforation-survived >65 days. The 30-day mortality rate following a bevacizumab-associated GI perforation was 50%. CONCLUSION: Bevacizumab-associated GI perforations in patients with recurrent ovarian carcinoma occurred in 4% of our patients. The prognosis of patients diagnosed with bevacizumab-associated GI perforations in this study was poor, and treatment should be individualized. PMID- 20004957 TI - Radical hysterectomy during the transition period from traditional to nerve sparing technique. AB - OBJECTIVE: The aim of the study was to compare peri- and postoperative data from patients operated on using the new nerve-sparing technique of radical hysterectomy with data gathered from those who underwent traditional radical hysterectomy. MATERIALS AND METHODS: A total of 20 patients with cervical cancer were included in the study. The study was carried out at a time when the authors had started to perform the nerve-sparing technique by using the descriptions from the literature. During the study period 10 patients underwent the nerve-sparing procedure while the other 10 patients underwent traditional radical hysterectomy. The two groups of patients were comparable in terms of mean age, body mass index, FIGO stage, and histological type; additionally, the follow-up period was similar for both groups. RESULTS: On the one hand, the mean total operative time (197.5+/ 51.4 vs. 155.5+/-39.6 min) and the mean time for the hysterectomy itself (154.5+/ 35.4 vs. 123.0+/-29.8 min) were significantly longer in the group operated on with nerve-sparing technique (p=0.05). Postoperatively, on the other hand, a post void residual urine volume of less than 50 ml was noted to occur significantly faster in the patients who had undergone the nerve-sparing technique (3.5+/-1.4 vs. 9.1+/-4.2 days, p=0.00078). CONCLUSIONS: Although during the introductory period nerve-sparing technique brings about an improvement in voiding function, it prolongs the total operative time in comparison to traditional radical hysterectomy. PMID- 20004958 TI - Upper abdominal surgery in advanced and recurrent ovarian cancer: role of diaphragmatic surgery. AB - OBJECTIVE: Upper abdominal spread of primary and recurrent ovarian cancer is often considered to be a major obstacle to achieve optimal residual disease at the end of surgery. In this study, we investigate the role of diaphragmatic debulking in the natural history of advanced and recurrent epithelial ovarian cancer patients, and the morbidity of this procedure according to clinico surgical characteristics. METHODS: Data from 234 consecutive patients with primary and recurrent advanced ovarian cancer, operated at Catholic University of Rome and Campobasso from January 1, 2005 and December 31, 2008, were retrospectively reviewed. RESULTS: Eighty-seven patients (37.2%) underwent a diaphragmatic surgery. Median age was 55 years (range 37-76). Diaphragmatic debulking was performed in 50 out of 120 patients at primary surgery (41.7%), in 16 out of 74 at interval debulking surgery (21.6%) and in 21 out of 40 secondary cytoreductions (52.5%). In the whole study population optimal residual disease at the end of surgery was achieved. The most frequent post-operative complication was pleural effusion, observed in 37 patients (42.5%). Presence of a post operative pleural effusion was correlated liver mobilization (52.3% vs. 16%; p<0.0027) and large diaphragmatic disease (>5 cm) removal (54.1% vs. 23.5%; p<0.034). CONCLUSIONS: Diaphragmatic surgery represents a crucial step in the debulking of advanced and recurrent ovarian cancer patients. Considering the natural history of advanced epithelial ovarian cancer and the rate of patients needing diaphragmatic debulking during primary cytoreduction, interval debulking surgery and secondary cytoreduction, this procedure should be present in the surgical repertoire of a gynecologic oncologist. PMID- 20004960 TI - Adhesion to tooth structure: a critical review of "macro" test methods. AB - OBJECTIVES: Bond strength between adhesive systems and the tooth structure is influenced by a large number of variables, which makes the comparison among studies virtually impossible. Also, failure often times propagates into the dental substrate or the composite, deeming the results questionable at best. In spite of the increased popularity gained by micro-tensile and micro-shear tests, in vitro evaluations using specimens with relatively large bonding areas remain frequent. This review focuses on aspects related to specimen geometry and test mechanics of "macro" shear and tensile bond strength tests. METHODS: Besides information drawn from the literature, the effect of some parameters on stress distribution at the bonded interface was assessed using finite element analysis (FEA). RESULTS: Bond strength tends to increase with smaller bonding areas and with the use of high elastic modulus composites. Stress concentration at the bonded interface is much more severe in shear compared to tension. Among shear methods, the use of the chisel shows the highest stress concentration. Within the limits suggested by the ISO/TS 11405, crosshead speed does not seem to influence bond strength values. Pooled data from currently available adhesives tested in either shear or tension showed 44% of adhesive failures, 31% mixed and 25% cohesive in the substrate (tooth or composite). A comparative bond strength study involving three adhesive systems revealed similarities between "macro" and "micro" counterparts regarding material ranking, whereas "macro" tests presented a higher incidence of cohesive failures. SIGNIFICANCE: Simplicity warrants "macro" bond strength tests an enduring popularity, in spite of their evident limitations. From a mechanical standpoint, knowing the stress distribution at the bonded interface and how it is affected by the materials and loading method used is key to explain the results. PMID- 20004959 TI - Sensory neuron-specific GPCR Mrgprs are itch receptors mediating chloroquine induced pruritus. AB - The cellular and molecular mechanisms mediating histamine-independent itch in primary sensory neurons are largely unknown. Itch induced by chloroquine (CQ) is a common side effect of this widely used antimalarial drug. Here, we show that Mrgprs, a family of G protein-coupled receptors expressed exclusively in peripheral sensory neurons, function as itch receptors. Mice lacking a cluster of Mrgpr genes display significant deficits in itch induced by CQ but not histamine. CQ directly excites sensory neurons in an Mrgpr-dependent manner. CQ specifically activates mouse MrgprA3 and human MrgprX1. Loss- and gain-of-function studies demonstrate that MrgprA3 is required for CQ responsiveness in mice. Furthermore, MrgprA3-expressing neurons respond to histamine and coexpress gastrin-releasing peptide, a peptide involved in itch sensation, and MrgprC11. Activation of these neurons with the MrgprC11-specific agonist BAM8-22 induces itch in wild-type but not mutant mice. Therefore, Mrgprs may provide molecular access to itch-selective neurons and constitute novel targets for itch therapeutics. PMID- 20004961 TI - My older sibling was drunk - younger siblings' drunkenness in relation to parental monitoring and the parent-adolescent relationship. AB - This study explored the links between having older siblings who get drunk, satisfaction with the parent-adolescent relationship, parental monitoring, and adolescents' risky drinking. Regression models were conducted based on a national representative sample of 3725 8th to 10th graders in Switzerland (mean age 15.0, SD = .93) who indicated having older siblings. Results showed that both parental factors and older siblings' drinking behaviour shape younger siblings' frequency of risky drinking. Parental monitoring showed a linear dose-response relationship, and siblings' influence had an additive effect. There was a non linear interaction effect between parent-adolescent relationship and older sibling's drunkenness. The findings suggest that, apart from avoiding an increasingly unsatisfactory relationship with their children, parental monitoring appears to be important in preventing risky drinking by their younger children, even if the older sibling drinks in such a way. However, a satisfying relationship with parents does not seem to be sufficient to counterbalance older siblings' influence. PMID- 20004962 TI - Identity status change during adolescence and young adulthood: a meta-analysis. AB - The present study was designed to examine developmental patterns of identity status change during adolescence and young adulthood through meta-analysis. Some 124 studies appearing in PsycINFO, ERIC, Sociological Abstracts, and Dissertation Abstracts International between 1966 and 2005 provided data. All calculations were performed using the software program, Comprehensive Meta-analysis. Results from longitudinal studies showed the mean proportion of adolescents making progressive identity status changes was .36, compared with .15 who made regressive changes and .49 who remained stable. Cross-sectional studies showed the mean proportion of moratoriums rising steadily to age 19 years and declining thereafter, while the mean proportion of the identity achieved rose over late adolescence and young adulthood; foreclosure and diffusion statuses declined over the high school years, but fluctuated throughout late adolescence and young adulthood. Meta-analyses showed that large mean proportions of samples were not identity achieved by young adulthood. Possible reasons for this phenomenon are explored. PMID- 20004963 TI - Molecular methods for pathogen detection in blood. PMID- 20004964 TI - Accurate and rapid identification of bacterial species from positive blood cultures with a DNA-based microarray platform: an observational study. AB - BACKGROUND: New DNA-based microarray platforms enable rapid detection and species identification of many pathogens, including bacteria. We assessed the sensitivity, specificity, and turnaround time of a new molecular sepsis assay. METHODS: 2107 positive blood-culture samples of 3318 blood samples from patients with clinically suspected sepsis were investigated for bacterial species by both conventional culture and Prove-it sepsis assay (Mobidiag, Helsinki, Finland) in two centres (UK and Finland). The assay is a novel PCR and microarray method that is based on amplification and detection of gyrB, parE, and mecA genes of 50 bacterial species. Operators of the test assay were not aware of culture results. We calculated sensitivity, specificity, and turnaround time according to Clinical and Laboratory Standards Institute recommendations. FINDINGS: 1807 of 2107 (86%) positive blood-culture samples included a pathogen covered by the assay. The assay had a clinical sensitivity of 94.7% (95% CI 93.6-95.7) and a specificity of 98.8% (98.1-99.2), and 100% for both measures for meticillin-resistant Staphylococcus aureus bacteraemia. The assay was a mean 18 h faster than was the conventional culture-based method, which takes an additional 1-2 working days. 34 of 3284 (1.0%) samples were excluded because of technical and operator errors. INTERPRETATION: Definitive identification of bacterial species with this microarray platform was highly sensitive, specific, and faster than was the gold standard culture-based method. This assay could enable fast and earlier evidence based management for clinical sepsis. PMID- 20004965 TI - Fighting overtreatment in adjuvant breast cancer therapy. PMID- 20004967 TI - Controlling the release of peptide antimicrobial agents from surfaces. AB - Medical conditions are often exacerbated by the onset of infection caused by hospital dwelling bacteria such as Staphylococcus aureus. Antibiotics taken orally or intravenously can require large and frequent doses, further contributing to the sharp rise in resistant bacteria observed over the past several decades. These existing antibiotics are also often ineffective in preventing biofilm formation, a common cause of medical device failure. Local delivery of new therapeutic agents that do not allow bacterial resistance to occur, such as antimicrobial peptides, could alleviate many of the problems associated with current antibacterial treatments. By taking advantage of the versatility of layer-by-layer assembly of polymer thin films, ponericin G1, an antimicrobial peptide known to be highly active against S. aureus, was incorporated into a hydrolytically degradable polyelectrolyte multilayer film. Several film architectures were examined to obtain various drug loadings that ranged from 20 to 150 microg/cm2. Release was observed over approximately ten days, with varying release profiles, including burst as well as linear release. Results indicated that film-released peptide did not suffer any loss in activity against S. aureus and was able to inhibit bacteria attachment, a necessary step in preventing biofilm formation. Additionally, all films were found to be biocompatible with the relevant wound healing cells, NIH 3T3 fibroblasts and human umbilical vein endothelial cells. These films provide the level of control over drug loading and release kinetics required in medically relevant applications including coatings for implant materials and bandages, while eliminating susceptibility to bacterial resistance. PMID- 20004968 TI - The recruitment of two consecutive and different waves of host stem/progenitor cells during the development of tissue-engineered bone in a murine model. AB - Angiogenesis plays a central role in bone regeneration, not only for the transport of nutrients, but also for locally directing skeletal stem/progenitor cells. Following ectopic implantation of porous ceramic cubes seeded with mouse GFP-labeled mesenchymal stem cells (MSC) into syngenic mice, we investigated the cascade of events leading to bone formation. Implants harvested at different times were enzymatically digested to generate single-cell suspensions. Recovered cells were sorted to separate GFP+implanted MSC and host recruited GFP- cells. We isolated and characterized two different waves of cells, migrating from the host to the MSC-seeded ceramic. The first migrated cell population, recovered 7 days after implantation, was enriched in CD31+endothelial progenitors, while the second one, recruited at day 11, was enriched in CD146+pericyte-like cells. Both populations were not recruited into the scaffold following implantation of a non MSC seeded ceramic. Pericyte-like cell mobilization was dependent on the first migrated endothelial cell population. Pericyte-like cells retained properties distinctive of stem cells, such as capacity of performing a high number of in vitro cell divisions and showed an osteogenic potential. Studies on the cross talk between implanted exogenous MSC and resident stem/progenitor cells could open new perspectives for future clinical applications. PMID- 20004966 TI - Adjuvant chemotherapy and timing of tamoxifen in postmenopausal patients with endocrine-responsive, node-positive breast cancer: a phase 3, open-label, randomised controlled trial. AB - BACKGROUND: Tamoxifen is standard adjuvant treatment for postmenopausal women with hormone-receptor-positive breast cancer. We assessed the benefit of adding chemotherapy to adjuvant tamoxifen and whether tamoxifen should be given concurrently or after chemotherapy. METHODS: We undertook a phase 3, parallel, randomised trial (SWOG-8814, INT-0100) in postmenopausal women with hormone receptor-positive, node-positive breast cancer to test two major objectives: whether the primary outcome, disease-free survival, was longer with cyclophosphamide, doxorubicin, and fluorouracil (CAF) given every 4 weeks for six cycles plus 5 years of daily tamoxifen than with tamoxifen alone; and whether disease-free survival was longer with CAF followed by tamoxifen (CAF-T) than with CAF plus concurrent tamoxifen (CAFT). Overall survival and toxicity were predefined, important secondary outcomes for each objective. Patients in this open-label trial were randomly assigned by a computer algorithm in a 2:3:3 ratio (tamoxifen:CAF-T:CAFT) and analysis was by intention to treat of eligible patients. Groups were compared by stratified log-rank tests, followed by Cox regression analyses adjusted for significant prognostic factors. This trial is registered with ClinicalTrials.gov, number NCT00929591. FINDINGS: Of 1558 randomised women, 1477 (95%) were eligible for inclusion in the analysis. After a maximum of 13 years of follow-up (median 8.94 years), 637 women had a disease free survival event (tamoxifen, 179 events in 361 patients; CAF-T, 216 events in 566 patients; CAFT, 242 events in 550 patients). For the first objective, therapy with the CAF plus tamoxifen groups combined (CAFT or CAF-T) was superior to tamoxifen alone for the primary endpoint of disease-free survival (adjusted Cox regression hazard ratio [HR] 0.76, 95% CI 0.64-0.91; p=0.002) but only marginally for the secondary endpoint of overall survival (HR 0.83, 0.68-1.01; p=0.057). For the second objective, the adjusted HRs favoured CAF-T over CAFT but did not reach significance for disease-free survival (HR 0.84, 0.70-1.01; p=0.061) or overall survival (HR 0.90, 0.73-1.10; p=0.30). Neutropenia, stomatitis, thromboembolism, congestive heart failure, and leukaemia were more frequent in the combined CAF plus tamoxifen groups than in the tamoxifen-alone group. INTERPRETATION: Chemotherapy with CAF plus tamoxifen given sequentially is more effective adjuvant therapy for postmenopausal patients with endocrine-responsive, node positive breast cancer than is tamoxifen alone. However, it might be possible to identify some subgroups that do not benefit from anthracycline-based chemotherapy despite positive nodes. FUNDING: National Cancer Institute (US National Institutes of Health). PMID- 20004969 TI - Effect of functionalised fluorescence-labelled nanoparticles on mesenchymal stem cell differentiation. AB - The combined use of nanoparticles and mesenchymal stem cells (MSC) in regenerative medicine requires the incorporation of the particles and, at the same time, undisturbed cell viability and maintenance of the multi-lineage potential of MSC. The aim of this study was to investigate the uptake of novel phosphonate-functionalised polystyrene nanoparticles prepared by miniemulsion polymerisation. After exposition of human MSC to the particles, their uptake and localisation were analysed by flow cytometry, confocal laser scanning microscopy (CLSM), and transmission electron microscopy (TEM). The osteogenic, adipogenic and chondrogenic differentiation potential was examined by analysing representative marker genes by RT-PCR. Flow cytometry revealed that after 5 and 16 days more than 98% of the MSC and of the cells, which underwent osteogenic and adipogenic differentiation were positive for particle association. CLSM and TEM demonstrated the successful intracellular incorporation of the particles without using any transfection agents and their presence over the cultivation period. The cell viability was found to be unaffected. Particle treated MSC maintained their potential for osteogenic, adipogenic and chondrogenic differentiation. It was concluded that the surface functionalisation with phosphonate groups provides a promising basis for the development of nanoparticles with high intracellular uptake rates for drug delivery or cell labelling. PMID- 20004970 TI - Anti-HIF-1alpha antibody-conjugated pluronic triblock copolymers encapsulated with Paclitaxel for tumor targeting therapy. AB - Targeted uptake of nanoscale controlled release polymer micelles encapsulated with drugs represents a potential powerful therapeutic technology. Herein we reported the development of anti-HIF-1alpha antibody-conjugated unimolecular polymer nano micelles filled with Paclitaxel for cancer targeting therapy. Pluronic triblock copolymers(Poly(ethylene glycol)-block-poly(propylene glycol) block-poly(ethylene glycol), PEO-block-PPO-block-PEO) P123 were functionalized with terminal carboxylic groups, and were characterized by infrared (IR) spectroscopy, nuclear magnetic resonance (NMR), thermogravimetric analysis (TGA), and differential scanning calorimetric (DSC). The amphiphilic copolymer nano micelles encapsulated with Paclitaxel were fabricated by self-assembly means, and then were conjugated with anti-HIF-1alpha antibody, the resultant anti-HIF-1alpha conjugated nano micelles filled with PTX (anti-HIF-1alpha-NMs-PTX nanocomposites) were characterized by dynamic light scattering (DLS) and transmission electron microscopy (TEM), and incubated with stomach cancer MGC-803 cells and HDF fibroblast cells, these treated cells were analyzed by MTT and cell-ELISA. The nanocomposites composed of anti-HIF-1alpha conjugated nano micelles filled with CdTe quantum dots were also prepared, and incubated with stomach cancer MGC-803 cells and HDF fibroblast cells for 24 h, then were observed by fluorescent microscope. Results showed that the anti-HIF-1alpha-NMs-PTX nanocomposites were successfully prepared, bound with stomach cancer MGC-803 cells specifically, were internalized, and released PTX inside cancer cells, and selectively killed cancer cells. In conclusion, unique anti-HIF-1alpha antibody-conjugated nano micelles filled with Paclitaxel can target and selectively kill cancer cells with over expression of HIF-1alpha, and has great potential in clinical tumor targeting imaging and therapy. PMID- 20004971 TI - Control of cell attachment through polyDNA hybridization. AB - Cell-cell interactions play vital roles in embryo development and in homeostasis maintenance. Such interactions must be stringently controlled for cell-based tissue engineering and regenerative medicine therapies, and methods for studying and controlling cell-cell interactions are being developed using both biomedical and engineering approaches. In this study, we prepared amphiphilic PEG-lipid polymers that were attached to polyDNA with specific sequences. Incubation of cells with the polyDNA-PEG-lipid conjugate transferred some of the polyDNA to the cells' surfaces. Similarly, polyDNA-PEG-lipid conjugate using polyDNA with a complementary sequence was introduced to the surfaces of other cells or to a substrate surface. Cell-cell or cell-substrate attachments were subsequently mediated via hybridization between the two complementary polyDNAs and monitored using fluorescence microscopy. PMID- 20004972 TI - The expression of caveolin-1 and the distribution of caveolae in the murine placenta and yolk sac: parallels to the human placenta. AB - The expression pattern of caveolin-1 and the distribution of caveolae in the murine placental labyrinth and visceral yolk sac have been determined. Immunoblot analysis demonstrates that both placenta and yolk sac express the protein caveolin-1. Immunofluorescence microscopy was used to determine which cell types in the placental labyrinth and yolk sac express caveolin-1. In yolk sac, detectable caveolin-1 was restricted to endothelial cells and smooth muscle cells of the vitelline vasculature and to mesothelial cells. Endoderm, the major cell type in the yolk sac, does not express caveolin-1 as assessed by this assay. In the labyrinth region of the placenta, endothelial cells express caveolin-1 but this protein was not detectable in any of the three trophoblast layers. These tissues were also examined by electron microscopy to determine which cell types contain the specialized plasma membrane microdomains known as caveolae. Morphologically detectable caveolae were present in endothelial and smooth muscle cells, as well as mesothelial cells of the yolk sac and in endothelial cells of the placental labyrinth. Neither endodermal cells of the yolk sac nor trophoblastic cells in the placental labyrinth contained caveolae-like structures. We conclude that caveolin-1 and caveolae have restricted distribution in the murine placenta and yolk sac and that this parallels the situation in human placenta. PMID- 20004973 TI - Differential effect of prolonged food restriction and fasting on hypothalamic malonyl-CoA concentration and expression of orexigenic and anorexigenic neuropeptides genes in rats. AB - Several lines of evidence suggest that malonyl-CoA in the hypothalamus plays an important role in monitoring and modulating body energy balance. In fasted state the level of malonyl-CoA concentration significantly decreases. Simultaneously, orexigenic neuropeptides (NPY - neuropeptide Y, AgRP - agouti-related peptide) genes are expressed at high level, whereas anorexigenic neuropeptides (CART - cocaine-and amphetamine-regulated transcript, POMC - proopiomelanocortin) genes are expressed at low level. When food intake resumes, opposite effect is observed. This study examined the effect of prolonged food restriction, common in humans trying to lose body weight on expression of orexigenic and anorexigenic neuropeptides genes and on malonyl-CoA content in rat whole hypothalamus. We observed an increase of NPY and AgRP mRNA levels in hypothalamus of rats kept on 30 days-long food restriction (50% of the amount of food consumed by controls). Simultaneously, a decrease of CART and POMC mRNA levels occurred. Refeeding caused a decrease in NPY and POMC mRNA levels without effect on AgRP and CART mRNA. Surprisingly, both prolonged food restriction and food restriction/refeeding caused the increase of malonyl-CoA level in whole hypothalamus. In contrast, fasting for 24h caused the decrease of malonyl-CoA level, which was associated with the up-regulation of NPY and AgRP genes expression and down-regulation of CART and POMC genes expression. After refeeding opposite effect was observed. These results indicate that prolonged food restriction and acute fasting, conditions in which energy expenditure exceeds intake, differentially affect malonyl-CoA concentration and similarly affect orexigenic and anorexigenic neuropeptide genes expression in whole rat hypothalamus. PMID- 20004974 TI - Preliminary molecular analysis of Clostridium difficile isolates from healthy horses in northern Italy. AB - Clostridium difficile, associated with a wide spectrum of diseases in humans, as well as in several animal species, is an important cause of colitis in adult horses and foals. The aim of this study was to investigate by toxin gene profile and PCR-ribotyping the molecular characteristics of 14 C. difficile strains isolated from 42 faeces of healthy horses. Both toxin genes, tcdA and tcdB, were present in only 1 isolate (7.1%). Six isolates (42.9%) demonstrated tcdA-/tcdB+ genotype, and seven isolates (50.0%) were tcdA-/tcdB-. All strains were binary toxin genes negative (cdtA-/cdtB-). The PCR-positive strains, except for the tcdA+/tcdB+ isolate, tested negative for, in vitro, A and/or B toxins production by EIA. Eleven distinct ribotypes were observed. In conclusion, C. difficile can be present in the normal intestinal flora of healthy adult horses, in addition to foals. These animals could therefore play an important role as potential reservoirs of toxigenic strains. PMID- 20004975 TI - Insulin potentiates FcepsilonRI-mediated signaling in mouse bone marrow-derived mast cells. AB - Factors contained in physiological microenvironments in tissues where mast cells differentiate and reside may influence mast cell responsiveness and modify antigen-dependent activation. A possible direct or indirect role of mast cell responses in diabetes mellitus prompted us to study the impact of insulin treatment on antigen triggered signaling pathways downstream of FcepsilonRI in bone marrow-derived mouse mast cells (BMMCs). We found that insulin alone stimulates tyrosine phosphorylation of tyrosine kinases Lyn, Syk, Fyn, the adapter protein Gab2 (Grb2-associated binding protein 2), Akt and activates ERK, JNK and p38 kinase. Effect of insulin on FcepsilonRI signaling pathways was marked by enhanced phosphorylation of Lyn, Fyn, Gab2 and Akt. Furthermore, BMMC stimulated with antigen in the presence of insulin responded with enhanced protein kinase theta (PKCtheta) activity and increased JNK phosphorylation when compared to BMMC triggered with antigen alone. Functional studies reveal enhanced degranulation and altered cytoskeletal rearrangement when BMMCs were treated simultaneously with insulin and antigen. Our results suggest that insulin tunes antigen-mediated responses of mast cells. PMID- 20004976 TI - A novel CXCL8 protein-based antagonist in acute experimental renal allograft damage. AB - Acute renal allograft damage is caused by early leukocyte infiltration which is mediated in part by chemokines presented by glycosaminoglycan (GAG) structures on endothelial surfaces. CXCL8 can recruit neutrophils and induce the firm arrest of monocytes on activated endothelial cells. A human CXCL8-based antagonist (dnCXCL8) designed to generate a dominant-negative mutant protein with enhanced binding to GAG structures and reduced CXCR1/2 receptor binding ability was tested in models of early allograft injury. The agent displayed enhanced binding to GAG structures in vitro and could antagonize CXCL8-induced firm adhesion of monocytes as well as neutrophils to activated microvascular endothelium in physiologic flow assays. In a rat model of acute renal damage, dnCXCL8 treatment limited proximal tubular damage and reduced granulocyte infiltration. In a Fischer 344 (RT1(lvl)) to Lewis (RT1(l)) rat acute renal allograft model, dnCXCL8 was found to reduce monocyte and CD8+ T-cell infiltration into glomeruli and to limit tubular interstitial inflammation and tubulitis in vivo. Early treatment of allografts with agents like dnCXCL8 may help reduce acute allograft damage and preserve renal morphology and thereby help limit chronic dysfunction. PMID- 20004977 TI - Unveiling patterns of affective responses in daily life may improve outcome prediction in depression: a momentary assessment study. AB - OBJECTIVE: Daily life affective responses are closely linked to vulnerability and resilience in depression. Prediction of future clinical course may be improved if information on daily life emotional response patterns is taken into account. METHOD: Female subjects with a history of major depression (n=83), recruited from a population twin register, participated in a longitudinal study using momentary assessment technology with 4 follow-up measurements. The effect of baseline daily life emotional response patterns (affect variability, stress-sensitivity and reward experience) on follow-up depressive symptomatology was examined. RESULTS: Both reward experience (B=-0.30, p=0.001) and negative affect variability (B=0.46, p=0.001) predicted future negative affective symptoms independent of all other dynamic emotional patterns and conventional predictors. CONCLUSION: Daily life information on dynamic emotional patterns adds to the prediction of future clinical course, independent of severity of symptoms and neuroticism score. Better prediction of course may improve decision-making regarding quantitative and qualitative aspects of treatment. PMID- 20004979 TI - Notch activation enhances IFNgamma secretion by human peripheral blood and decidual NK cells. AB - NK cells specialize in killing tumor cells and virally infected cells and also possess non-cytotoxic functions, which include secretion of a variety of cytokines and growth factors. Their activity is mediated by a vast repertoire of inhibitory and activating NK receptors. Recently, it was demonstrated that ligation of the Notch receptor plays a significant role not only in T cell development but also in human T cell and mouse NK cell activation. However, the involvement of Notch triggering in human NK cell activity has not yet been determined. Here we show that Notch1 and Notch2, but not Notch3 and Notch 4, are expressed in human peripheral blood NK cells and in decidual NK cells. We demonstrate that in peripheral blood NK cells only the Notch ligand Delta4 could interact with Notch, whereas in decidual NK cells both Delta1 and Delta4 can interact with Notch. Finally, we show that Notch activation in these cells leads to increased secretion of IFNgamma. We therefore present here a new function of the Notch receptors as enhancers of peripheral blood NK cell and decidual NK cell functions. PMID- 20004980 TI - Attitudes towards long-acting depot antipsychotics: a survey of patients, relatives and psychiatrists. AB - In many countries fewer than 20% of individuals with schizophrenia receive depot antipsychotic medication. Frequently stated reasons are psychiatrist's, patient's and relative's objections to depot treatment. This is the first study that directly compares the attitudes to depot antipsychotics of psychiatrists, patients and relatives. A semi-structured questionnaire about their attitudes towards depot antipsychotics was completed by 255 participants (83 patients diagnosed with schizophrenic disorder, 81 psychiatrists in private practice and 91 relatives, not directly related to the patients). Patients were more negative towards depot injections than psychiatrists and relatives. They particularly fear to be constricted in their autonomy when treated with depot antipsychotics and that injections might be painful. About 67% of all patients in our sample did not receive information about depot antipsychotics from their psychiatrist. Less than 10% of psychiatrists offer depot treatment after a first psychotic episode. Psychiatrists use depot antipsychotics in a conservative way, although they attribute positive traits to the method. Patients' negative attitudes might relate to the low level of information. To enhance the use of depot antipsychotics, information practices of psychiatrists should be improved. Patients should be informed about different forms of treatment during early stages of the illness. PMID- 20004978 TI - Genetic and environmental influences on psychiatric comorbidity: a systematic review. AB - BACKGROUND: The purpose of this review is to systematically appraise the peer reviewed literature about the genetic and environmental determinants of psychiatric comorbidity, focusing on four of the most prevalent types of psychopathology: anxiety disorders, depression, conduct disorder and substance abuse. METHODS: We summarize existing empirical research on the relative contribution that genetic, nonshared and shared environmental factors make to the covariance between disorders, and evidence about specific genes and environmental characteristics that are associated with comorbidity. RESULTS: Ninety-four articles met the inclusion criteria and were assessed. Genetic factors play a particularly strong role in comorbidity between major depression and generalized anxiety disorder or posttraumatic stress disorder, while the non-shared environments make an important contribution to comorbidity in affective disorders. Genetic and non-shared environmental factors also make a moderate-to strong contribution to the relationship between CD and SA. A range of candidate genes, such as 5HTTLPR, MAOA, and DRD1-DRD4, as well as others implicated in the central nervous system, has been implicated in psychiatric comorbidity. Pivotal social factors include childhood adversity/life events, family and peer social connections, and socioeconomic and academic difficulties. LIMITATIONS: Methodological concerns include the use of clinical case-control samples, the focus on a restricted set of individual-level environmental risk factors, and restricted follow-up times. CONCLUSIONS: Given the significant mental health burden associated with comorbid disorders, population-based research on modifiable risk factors for psychiatric comorbidity is vital for the design of effective preventive and clinical interventions. PMID- 20004981 TI - Antiviral effect of recombinant equine interferon-gamma on several equine viruses. AB - Recombinant equine interferon-gamma (reIFN-gamma) was prepared using a baculovirus expression system and its antiviral activity was investigated using several equine viruses. The reIFN-gamma suppressed the replication of all equine viruses used in the present experiment in horse cell cultures, but did not affect the growth of host cells at concentrations of less than 1000 u/ml. A strong antiviral effect was observed, especially against RNA viruses. Equine picornavirus, equine rhinovirus and equine arteritis virus could not be propagated at all in 100 u/ml reIFN-gamma when 100 TCID(50) of infective viruses was inoculated to cultivated horse cells. DNA viruses, equine herpesvirus types 1, 2, 3 and 4 and equine adenovirus, were less sensitive to reIFN-gamma but their growth became less than 1/100 in the cells treated with 100 u/ml reIFN-gamma compared to untreated cells. The antiviral effects were decreased in the cells of heterologous species and more than 1000 u/ml reIFN-gamma was required to induce an antiviral effect. PMID- 20004982 TI - Ryks: new partners for Wnts in the developing and regenerating nervous system. AB - Conserved Ryk transmembrane proteins, tyrosine kinase-related Wnt receptors, are important during neurogenesis, axon guidance and synaptogenesis. Here, we review the increasingly complex biology of the Wnt/Ryk pathway, emphasizing the mechanisms by which Ryks transduce or sometimes block the Wnt signal. Recent studies reveal that Wnts signal through Ryk via multiple mechanisms, including nuclear translocation of their intracellular domains and pathways employing Src Family Kinases and members of the canonical Wnt pathway. We also discuss reports indicating that Wnt/Ryk axon guidance roles are evolutionarily conserved and Wnt/Ryk interactions are required for motoneuron target selection and synaptogenesis at the neuromuscular junction. Recent findings that injury-induced Wnt/Ryk pathway activation inhibits axon regeneration underscore the importance of further understanding this novel pathway. PMID- 20004983 TI - Induction of hepatic carbonyl reductase/20beta-hydroxysteroid dehydrogenase mRNA in rainbow trout downstream from sewage treatment works--possible roles of aryl hydrocarbon receptor agonists and oxidative stress. AB - Carbonyl reductase/20beta-hydroxysteroid dehydrogenase (CR/20beta-HSD) serves both as a key enzyme in the gonadal synthesis of maturing-inducing hormone in salmonids, and as an enzyme protecting against certain reactive oxygen species. We have previously shown that mRNA of the hepatic CR/20beta-HSD B isoform is increased in rainbow trout caged downstream from a Swedish sewage treatment plant. Here, we report an increase of both the A as well as B form in fish kept downstream from a second sewage treatment plant. The two mRNAs were also induced in fish hepatoma cells in vitro after exposure to effluent extract. This indicates that the effects observed in vivo could be a direct effect on the liver, i.e. the mRNA induction does not require a signal from any other organ. When fish were exposed in vivo to several effluents treated with more advanced methods (ozone, moving bed biofilm reactor or membrane bioreactor) the expression of hepatic mRNA CR/20beta-HSD A and B was significantly reduced. Their abundance did not parallel the reduction of estrogen-responsive transcripts, in agreement with our previous observations that ethinylestradiol is not a potent inducer. Treatment with norethisterone, methyltestosterone or hydrocortisone in vivo did not induce the hepatic CR/20beta-HSD A and B mRNA expression. In contrast, both isoforms were markedly induced by the aryl hydrocarbon receptor agonist beta naphthoflavone as well as by the pro-oxidant herbicide paraquat. We hypothesize that the induction of CR/20beta-HSD A and B by sewage effluents could be due to anthropogenic contaminants stimulating the aryl hydrocarbon receptor and/or causing oxidative stress. PMID- 20004984 TI - Co-tolerance of phytoplankton communities to photosynthesis II inhibitors. AB - Natural variability in sensitivity and pollution induced community tolerance (PICT) to atrazine, isoproturon and diuron and a mixture of these three herbicides to natural algal assemblages in mesocosms was determined. The specificity of PICT was examined by evaluating co-tolerance pattern for these photosystem-II (PSII) inhibitors. Phytoplankton communities were constantly exposed to equipotent concentrations of atrazine, isoproturon, diuron namely the 30% hazard concentration (HC(30)) obtained from species sensitivity distributions and an equitoxic mixture (Sigma3 x 1/3 x HC(30) of each herbicide) for five weeks in outdoor mesocosms. Induction of tolerance to the various herbicides was investigated by photosynthetic efficiency measurements of the algal assemblages in short-term laboratory tests. The composition of the algal communities in the various treatments was determined and ordination techniques such as the principal component analysis (PCA) were applied to log-transformed data to compare the seasonal community structure development. Temporal variation in sensitivity of the control algal assemblage to atrazine and isoproturon, but less to diuron was observed. The results further demonstrated that the control communities were in general more sensitive than the treated ones over the whole period tested indicating an enhanced tolerance of pre-exposed phytoplankton in the mesocosms. Co-tolerance was also observed for atrazine pre-exposed algal community to isoproturon, however, not vise versa. A pre-exposure to diuron induced similar tolerance to all three herbicides. A pre-exposure to the mixture treatment also lead to tolerance to isoproturon and diuron, less to atrazine. Overall, the observed co-tolerance pattern indicates that co-tolerance was not comparable between the herbicides with strong similarity in their biochemical mode of action. PMID- 20004985 TI - Diagnostic accuracy of dual-source computed tomography in the characterization of coronary atherosclerotic plaques: comparison with intravascular optical coherence tomography. AB - BACKGROUND: Dual-source computed tomography (DSCT) has enabled us to non invasively visualize coronary artery stenosis, but its ability to characterize coronary atherosclerotic plaques (ASPs) has not been evaluated. Intravascular optical coherence tomography (OCT) provides tissue images of coronary artery wall that are validated by pathohistological studies. We studied the diagnostic accuracy of DSCT in the characterization of coronary ASPs, especially lipid-rich ASP with thin fibrous cap (TCFA), in comparison with OCT. METHODS: DSCT and OCT were used to image non-stenotic ASPs in non-culprit coronary arteries of 17 acute coronary syndrome (ACS) patients, and 162 coronary regions were enrolled. RESULTS: The mean CT values of fibrous ASP, ASP with lipid core, and ASP with calcium deposit were 77.5, 28.9, and 515.9 HU, respectively (P<0.0001). ASP with calcium deposit was detected with a sensitivity of 88.9% and a specificity of 98.6%, while ASP with lipid core was detected by DSCT with a relatively low sensitivity of 73.1% and a high specificity of 94.0%. In TCFA, cross-sectional areas of both ASP and lipid core were significantly larger, mean CT value of ASP was significantly lower, and concomitant calcification was more frequently observed compared with lipid-rich ASP with thick fibrous cap (ThCFA). The combination of these CT parameters seems to be a useful index for the differentiation of TCFA from ThCFA. CONCLUSION: DSCT is useful for non-invasive evaluation of calcified and fibro-fatty tissue characters in coronary artery plaque, but it is still not able to differentiate TCFA, one of the features of vulnerable plaque. PMID- 20004986 TI - A higher level of IgM anti-oxidized LDL antibodies is associated with a lower severity of coronary atherosclerosis in patients on statins. AB - BACKGROUND: Several studies show the association between anti-oxidized LDL antibodies and atherosclerosis. A certain association also seems to exist between the levels of these antibodies and treatment with statins. However, the clinical importance of these autoantibodies is still under discussion. The aim of this study was to determine whether levels of anti-oxidized LDL antibodies are associated with the presence of multivessel coronary artery disease according to whether statins were or were not being taken. METHODS AND RESULTS: The study included 236 patients who underwent invasive coronary angiography with quantification of coronary atherosclerotic lesions to determine the presence of multivessel coronary artery disease. Measurements were made of IgG and IgM anti oxidized LDL antibodies. In the group of patients who were taking statins, the percentage of patients with IgM anti-oxidized LDL antibodies below 50th percentile was significantly higher in those with multivessel coronary artery disease (57.4% vs. 37.5%, P<0.05). A logistic regression model in the patients taking statins showed that the variable significantly associated with the presence of multivessel coronary artery disease was the level of IgM anti oxidized LDL antibodies. CONCLUSIONS: The severity of multivessel coronary artery disease was inversely related with the levels of IgM anti-oxidized LDL antibodies, especially in the patients on statins. PMID- 20004987 TI - Initial Q waves and outcome after reperfusion therapy in patients with ST elevation acute myocardial infarction: a systematic review. AB - BACKGROUND: Patients with ST elevation acute myocardial infarction (STEMI) have different outcome depending on the reperfusion strategy. METHODS: To discern if the presence of initial Q waves in the infarct leads is a useful prognostic parameter in STEMI patients within 6 h of symptom onset treated by different reperfusion strategies (fibrinolysis, fibrinolysis followed by percutaneous coronary intervention [PCI], and primary PCI) we performed a systematic review on outcome comparing patients with and without initial Q waves. RESULTS: The relative risks for those with Q waves were significantly raised for both mortality and the composite outcome of mortality, congestive heart failure or cardiogenic shock, and at both 30-day and 90-day time points. The relative risk for mortality varied from 2.18 (95% CI 1.32-3.61) at 30 days to 2.54 (95% CI 1.87 3.44) at 90 days. The relative risk for composite outcome was 2.28 (95% CI 1.71 3.04) at 30 days and 2.25 (95% CI 1.81-2.80) at 90 days. CONCLUSION: The presence of initial Q waves is a relatively robust parameter to stratify outcome regardless of the reperfusion methods. PMID- 20004989 TI - A simulation study to assess statistical methods for binary repeated measures data. AB - Binary repeated measures data are commonly encountered in both experimental and observational veterinary studies. Among the wide range of statistical methods and software applicable to such data one major distinction is between marginal and random effects procedures. The objective of the study was to review and assess the performance of marginal and random effects estimation procedures for the analysis of binary repeated measures data. Two simulation studies were carried out, using relatively small, balanced, two-level (time within subjects) datasets. The first study was based on data generated from a marginal model with first order autocorrelation, the second on a random effects model with autocorrelated random effects within subjects. Three versions of the models were considered in which a dichotomous treatment was modelled additively, either between or within subjects, or modelled by a time interaction. Among the studied statistical procedures were: generalized estimating equations (GEE), Marginal Quasi Likelihood, likelihood based on numerical integration, penalized quasi likelihood, restricted pseudo likelihood and Bayesian Markov Chain Monte Carlo. Results for data generated by the marginal model showed autoregressive GEE to be highly efficient when treatment was within subjects, even with strongly correlated responses. For treatment between subjects, random effects procedures also performed well in some situations; however, a relatively small number of subjects with a short time series proved a challenge for both marginal and random effects procedures. Results for data generated by the random effects model showed bias in estimates from random effects procedures when autocorrelation was present in the data, while the marginal procedures generally gave estimates close to the marginal parameters. PMID- 20004990 TI - A stochastic mathematical model of the within-herd transmission dynamics of Porcine Reproductive and Respiratory Syndrome Virus (PRRSV): fade-out and persistence. AB - A stochastic, mathematical model of a farrow-finish pig herd was developed and used to investigate the within-herd transmission dynamics of PRRSV, and to examine patterns of on-farm persistence and fade-out. The model was structured to represent the management of a typical European pig herd. Three parameters determining the natural history of infection were derived from the literature. Transmission parameters were chosen using PRRSV antibody data from a cross sectional study of 103 pig herds (Evans et al., 2008). The seroprevalence by age was generated from the model at 21-day intervals and was compared to the cross sectional field data using log-likelihood, accounting for the accuracy of the ELISA test used. The model was run for various isolation practices of purchased gilts, contact structure, herd size and the frequency of re-introduction of infectious gilts. The time-dependent log-likelihood patterns varied between herds in a similar way to patterns observed from serological values from the 103 farms. Essentially they indicated two patterns of seroprevalence: herds in which PRRSV was stably persistent, and herds in which PRRSV was unstable, either recently introduced or recently faded-out. With a herd size of 327 sows with identical management, fade-out of virus occurred within 4 weeks in 21.9% of simulations. Without isolation of gilts from sows, fade-out within 250 days decreased from 81.6% to 14.3% and for herd sizes of 75, 150, 300 and 600, the probability of persistence of virus for >1200 days was 4%, 13.4%, 20.4% and 18.2%, respectively. Introduction of virus at a rate of approximately 0.37 times per year resulted in virus persisting for >1200 days in 32.4% of simulations, compared with 17.6% for no re-introduction. Fade-out of virus was most likely to occur within breeding females before virus reached young stock. Persistence was more likely once PRRSV was present in piglets which in turn infected rearing-pigs. The probability of persistence was higher with increased herd size, increased contact between different age groups and increased re-introduction of infectious gilts. The ability of the model to capture the variability in cross-sectional, age-related serological patterns suggests that the processes of re-introduction, persistence and fade-out of PRRSV play critical roles in PRRSV epidemiology. The potential importance to pig production and transmission of virus between herds is discussed. PMID- 20004991 TI - Who will care? Employment participation and willingness to supply informal care. AB - The impact of informal care responsibilities on the willingness and ability of caregivers to undertake paid employment has been the subject of a number of studies. In contrast, the effect of employment status on willingness to undertake informal care has been less well explored. This paper concentrates on this less studied direction of causality using the data provided by 15 waves of the British Household Panel Survey. We find that employment participation and earnings both impact negatively on willingness to supply informal care. This evidence has implications for health and social care policy since informal care has been shown to be a significant substitute for formal long-term care. PMID- 20004992 TI - Forisomes: calcium-powered protein complexes with potential as 'smart' biomaterials. AB - Sieve tubes in legumes contain forisomes, which are spindle-like bodies that are composed of ATP-independent, mechanically active proteins. Upon injury, forisomes occlude sieve tubes by dispersion and thus, help to prevent loss of nutrient-rich transport sap. Forisome enlargement by dispersion is brought about by Ca2+ induced conformational changes that confer radial expansion and longitudinal contraction. Forisomes recontract upon Ca2+ removal. In vitro, forisomes reversibly disperse and contract in the presence or absence of Ca2+, respectively, and at distinct pHs. Recently, forisomes have received renewed attention because of their unique capacity to convert chemical into mechanical energy independent of high-energy organic compounds. Forisome-based 'smart' materials can be used to produce self-powered monitoring and diagnostic systems. Here, we focus on physiological, chemical and physical aspects of forisomes and discuss their potential as biomimetic devices. PMID- 20004993 TI - Inhibition of Staphylococcus aureus in broth and meat broth using synergies of phenolics and organic acids. AB - This study assessed the occurrence of an enhancing inhibitory effect of the combined application of thymol and carvacrol with lactic and acetic acid against Staphylococcus aureus using the determination of Fractional Inhibitory Concentration (FIC) index and kill-time assay in meat broth and in a food model (meat). Minimum Inhibitory Concentration (MIC) of carvacrol and thymol were 1.25 and 0.6 microL/mL, respectively. FIC indices of the combined application of the phenolic constituents and organic acids were 0.5 suggesting a synergic interaction. No difference (p>0.05) was found among the bacterial counts for the meat broth added of the phenolics alone or in combination with lactic acid at sub inhibitory concentrations. Contrarily, the bacterial counts found for the meat broth added of the phenolics alone were significantly lower (p<0.05) than the counts obtained for the broth added of the combination of phenolics and acetic acid. No difference (p>0.05) was found among the bacterial counts for the meat model added of phytochemicals alone and in combination with the organic acids. The tested antimicrobials alone or in mixture provided smaller anti staphylococcal effect in meat broth than in meat model. These results could arise as an interesting approach for the improvement of food preservation using more natural procedures, considering the current demand of consumer and sensory quality of foods. PMID- 20004994 TI - Genetic Fusarium chemotyping as a useful tool for predicting nivalenol contamination in winter wheat. AB - Fusarium graminearum [teleomorph Gibberella zeae] and Fusarium culmorum together with Fusarium poae are the main species known to produce nivalenol (NIV). The NIV content in wheat (Triticum aestivum L.) harvested in Luxembourg was investigated in 2007 and 2008 at 17 different locations. Species determination and genetic chemotyping of F. graminearum and F. culmorum were used to understand the spatial distribution of NIV producers in wheat from Luxembourg. Three hundred thirteen F. graminearum, 175 F. culmorum and 117 F. poae strains respectively were isolated. Chemotypes of the first two species were determined by PCR and confirmed on a sub sample of single isolates by LC-MS/MS analysis. The 15-acetylated DON chemotype of F. graminearum was dominant in both years representing 94.2% of the population while the NIV chemotype represented 5.8%. The F. culmorum chemotypes were rather evenly distributed, with 3-acetylated DON and NIV profiles present with similar abundances (53.2% and 46.8%, respectively). NIV presence in wheat flour obtained from the 17 sites was correlated with the number of F. culmorum (NIV chemotype) isolated from 100 seeds, suggesting its primary role in NIV production on grains. The predictive power for identifying NIV contamination in grains based on NIV chemotype presence was confirmed by coupling the isolation procedure with a cut off value, resulting in the successful identification (100%, p=0.008) of NIV contamination in grains collected from 9 additional experimental sites. In conclusion, the results highlight the importance of chemotyping for improved prediction of toxin contamination in wheat. PMID- 20004995 TI - Physician scarcity is a predictor of further scarcity in US, and a predictor of concentration in Japan. AB - OBJECTIVES: To assess the effects of geographic diffusion of physicians from medically oversupplied toward undersupplied areas driven by economic competition among physicians and political interventions in Japan and US. METHODS: A quantitative evaluation of physician workforce changes at the community level between 1980 and 2005, using municipality-based (Japan) and county-based (US) census data. RESULTS: The overall number of physicians per 100,000 population (physician-to-population ratio: PPR) increased from 130 to 203 in Japan and 158 to 234 in US. In this context, a higher proportion (30.1%) of the quintile communities with lowest PPRs in 1980 has further decreased their PPRs in US than in Japan (21.6% in 2005). In multivariate analysis low PPR was a positive predictor of PPR decrease in the US communities (odds ratio 1.26; 95% confidence interval 1.01-1.58), while it was a negative predictor in Japanese communities (0.69; 0.57-0.83). CONCLUSIONS: Physician scarcity is associated with further scarcity in US communities, while scarcity is associated with recovery from scarcity in Japanese communities. Competition-based physician diffusion strategies and various interventions to address the maldistribution of physicians apparently have not worked effectively in US compared with Japan. PMID- 20004996 TI - Stakeholder perceptions of aid coordination implementation in the Zambian health sector. AB - In this study, we analysed stakeholder perceptions of the process of implementing the coordination of health-sector aid in Zambia, Africa. The aim of coordination of health aid is to increase the effectiveness of health systems and to ensure that donors comply with national priorities. With increases in the number of donors involved and resources available for health aid globally, the attention devoted to coordination worldwide has risen. While the theoretical basis of coordination has been relatively well-explored, less research has been carried out on the practicalities of how such coordination is to be implemented. In our study, we focused on potential differences between the views of the stakeholders, both government and donors, on the systems by which health aid is coordinated. A qualitative case study was conducted comprising interviews with government and donor stakeholders in the health sector, as well as document review and observations of meetings. Results suggested that stakeholders are generally satisfied with the implementation of health-sector aid coordination in Zambia. However, there were differences in perceptions of the level of coordination of plans and agreements, which can be attributed to difficulties in harmonizing and aligning organizational requirements with the Zambian health-sector plans. In order to achieve the aims of the Paris Declaration; to increase harmonization, alignment and ownership--resources from donors must be better coordinated in the health sector planning process. This requires careful consideration of contextual constraints surrounding each donor. PMID- 20004997 TI - Value of quantitative analysis of circulating cell free DNA as a screening tool for lung cancer: a meta-analysis. AB - OBJECTIVE: Quantitative analysis of circulating cell free DNA is considered as a possible aid for lung cancer screening. We aimed to comprehensively review the evidence for use of circulating cell free DNA to screen for lung cancer. METHODS: After a systematic review of English language studies, sensitivity, specificity, and other measures of accuracy of circulating DNA assay in the diagnosis of lung cancer were pooled using random-effects models. Summary receiver operating characteristic curves were used to summarize overall test performance. RESULTS: Ten studies met our inclusion criteria. The summary estimates for quantitative analysis of circulating cell free DNA in lung cancer screening in the studies included were as follows: sensitivity, 0.80 (95% confidence interval (CI), 0.77 0.83); specificity, 0.77 (95% CI, 0.74-0.80); positive likelihood ratio, 4.54 (95% CI, 2.66-7.76); negative likelihood ratio, 0.28 (95% CI, 0.19-0.40); and diagnostic odds ratio, 20.33 (95% CI, 10.12-40.86). CONCLUSIONS: The current evidence suggests that the diagnostic accuracy of quantitative analysis of circulating DNA is not lower than conventional serum biomarkers for lung cancer screening, at least. However, it is not recommend for lung cancer screening alone, because its discrimination power is not very perfect. The value of circulating DNA assay in combination with conventional markers for lung cancer detection deserved further investigation. PMID- 20004998 TI - ATM polymorphisms and risk of lung cancer among never smokers. AB - The ataxia-telangiectasia mutated (ATM) gene, an important caretaker of overall genome stability, is thought to play a role in the development of human malignancy. Therefore, we hypothesized that sequence variants in ATM may influence the disposition to lung cancer. In this hospital-based matched case control study, nine ATM single nucleotide polymorphisms (rs189037, rs228597, rs228592, rs664677, rs609261, rs599558, rs609429, rs227062, and rs664982) were genotyped in 730 lung cancer patients and 730 healthy controls. Pairwise linkage disequilibrium among nine polymorphisms in the ATM gene was very high. None of the main effects of any of the ATM polymorphisms were related to the risk of lung cancer. Interestingly, ATM polymorphisms were significantly associated with lung cancer among never smokers, and the association was modulated by low-level exposure to carcinogens such as environmental tobacco smoke. When the haplotypes of nine ATM polymorphism sites were studied, no overall association between ATM haplotypes and risk of lung cancer was found. However, the frequency distribution of haplotypes between lung cancer cases and controls was significant in the never smokers (P=0.009), demonstrating that haplotypes have a significant effect on the risk of lung cancer. In conclusion, we found that never smokers with sequence variants of the ATM gene may be at increased risk for lung cancer. Our data also suggest this association may be further modified by exposure to environmental tobacco smoke. This study suggests support to the literature that ATM polymorphisms and environmental tobacco smoke exposure have a role in lung carcinogenesis among never smokers. PMID- 20004999 TI - Source-specific effects of micronutrients in lung cancer prevention. AB - The role of micronutrients in lung cancer prevention is controversial, as observational and experimental studies have generated contradicting results. These discrepancies between studies may be due to different effects of micronutrients depending on source (diet or supplements). The objective of this study was to evaluate the association between vitamin C, E, folate and beta carotene and lung cancer risk while focusing on source-specific effects of dietary and supplemental intake. The association was evaluated in a cohort of 55,557 Danes who completed a food frequency questionnaire including information on consumption of vitamin C, E, folate and beta-carotene from diet and supplements. Incidence rate ratios of lung cancer were calculated using Cox proportional hazards models. During a median follow-up of 10.6 years, 721 incident lung cancer cases were diagnosed. We found a significant protective effect of dietary vitamin E intake and a significantly higher lung cancer risk with supplemental beta-carotene and dietary folate intake. All three micronutrients exhibited significant source-specific effects. The harmful effect of dietary folate is, however, most likely to be due to uncontrolled confounding. Our results indicate source-specific effects of vitamin E and beta-carotene in lung cancer prevention with a preventive effect of dietary vitamin E and a harmful effect of supplemental beta-carotene. Future studies on micronutrients and lung cancer should take source into account. PMID- 20005001 TI - Differential T-cell responses and allergen uptake after exposure of dendritic cells to the birch pollen allergens Bet v 1.0101, Bet v 1.0401 and Bet v 1.1001. AB - The major birch pollen allergen Bet v 1 is present in pollen as a mixture of at least 14 isoforms that share high sequence and structural identities. These isoforms possess either a high or a low IgE-binding capacity which defines them as allergenic or hypoallergenic. Recently, we could demonstrate that only the allergenic isoform Bet v 1.0101 was able to induce an IgE response in birch pollen allergic individuals. The hypoallergenic isoforms Bet v 1.0401 and Bet v 1.1001 were unable to induce IgE synthesis. T-helper cell responses against allergens are characterised by increased levels of Th2 cytokines. Therefore, we examined extent and polarisation of the Th cell response and the kinetics of the allergen uptake after exposure of dendritic cells (DCs) to these isoforms. Monocyte-derived DCs (MDDCs) from birch pollen allergic and non-atopic individuals stimulated with Bet v 1.0101, Bet v 1.0401 or Bet v 1.1001 in combination with the maturation factors TNF-alpha and IL-1beta resulted in a mature DC phenotype as measured by costimulatory molecule up-regulation. Only Bet v 1.0101-stimulated MDDCs from allergic subjects enhanced proliferation of autologous Th cells and the expression of the Th2 cytokines IL-5 and IL-13. Immature MDDCs of allergic individuals internalised equivalent amounts of the allergenic Bet v 1.0101 and the hypoallergenic Bet v 1.0401. In contrast, the uptake of the hypoallergenic Bet v 1.0401 by immature MDDCs of non-atopic individuals was significantly higher. These results provide evidence that DCs discriminate between allergens and highly related hypoallergens. This process may have an impact on the early phase of sensitisation. PMID- 20005002 TI - Leaf proteome responses of Arabidopsis thaliana exposed to mild cadmium stress. AB - The leaf proteome of 3-week-old Arabidopsis thaliana seedlings exposed for 1 week to low, environmentally realistic Cd concentrations was investigated. The data indicated that at 1muMCd, A. thaliana plants adapted their metabolism to cope with the Cd exposure. As a result, only moderate protein changes were observed. However, at 10muMCd, severe stress was indicated by growth reduction and chlorosis of rosette leaves at the macroscopic level and by lipid peroxidation and enhanced peroxidase activity at the cellular level. Of the 730 reproducible proteins among all gels, 21 were statistically upregulated in response to Cd. These proteins can be functionally grouped into 5 classes: proteins involved in (1) oxidative stress response, (2) photosynthesis and energy production, (3) protein metabolism, (4) gene expression and finally, (5) proteins with various or unknown function. In order to provide greater insight into the mechanisms induced on Cd exposure, a working model is proposed. PMID- 20005003 TI - NAM-1gene polymorphism and grain protein content in Hordeum. AB - Grain protein content (GPC) is a key quality factor for malting and brewing process. In wheat, a QTL explaining a large part of GPC variation was identified, which co-localizes with a gene encoding a NAC transcription factor (TtNAM-B1). NAC transcription factors influence GPC by their role in the regulation of senescence and in protein remobilization. An orthologous gene was discovered on barley chromosome 6H where a GPC QTL was mapped. In this study, we identify allelic variation of the NAM-1 gene for three species of Hordeum representing wild and cultivated barley and we investigate the possible link with GPC. Three haplotypes were identified, one corresponds to the sequences of 11 European varieties representing H. vulgare, one corresponds to the sequence found in H. spontaneum and one represents the sequence of H. bulbosum. Three SNPs were identified between H. spontaneum sequence and H. vulgare sequence. One of the H. bulbosum polymorphisms leads to the introduction of a stop codon and a non functional protein. Differences in GPC between the 11 varieties were found but no polymorphism in the NAM-1 gene was observed, suggesting that differences in expression of the HvNAM-1 gene or other genes should play a role in GPC regulation. Nevertheless based on published values for GPC of H. bulbosum and H. spontaneum compared to GPC measured here in H. vulgare, the non-functional protein is associated with the lower GPC, suggesting that loss of functionality of the NAM-1 gene in Hordeum is related to lower GPC. Moreover H. spontaneum GPC seems to be higher than H. vulgare GPC, suggesting also that allelic variation of the functional NAM-1 gene could be associated with GPC variation within the genus Hordeum. PMID- 20005004 TI - [CINCA syndrome: a rare cause of papilledema. The case of homozygous twins]. AB - CINCA syndrome is an autoinflammatory disease in childhood characterized by multisystemic manifestations: cutaneous, articular, and neurological including sensory organs. We report the case of homozygous twins affected by CINCA syndrome. The diagnosis was evoked on the basis of multiple systemic symptoms (multiple episodes of fever of unknown origin, mental retardation, short stature, meningitis, hearing loss, bilateral papilledema) and confirmed by the presence of a CIAS1 mutation on genetic analysis. After few months of treatment by anakinra (an interleukin-1 receptor antagonist) the children began to grow again and we noted regression of the biological inflammatory syndrome. PMID- 20005005 TI - [Analysis of a new visual field index, the VFI, in Ocular Hypertension and Glaucoma]. AB - OBJECTIVE: The Visual Field Index -VFI- is a new perimetric index allowing evaluation of visual function level and progression. In this study, we analysed this new index, that provides a trend analysis of visual field loss progression in Ocular Hypertension and Glaucoma. We also compared results with event analysis. METHODS: Retrospective study on 94 eyes of 54 patients: 35 OHT(ocular hypertension), 34 early POAG (primary open angle glaucoma) (0>MD>-6 dB), 13 moderate POAG (-6>MD>-12 dB) and 12 advanced POAG (MD<-12 dB), with a mean follow up of 6.5 years (4 to 8 years). Each subject performed a mean number of 10 standard automated perimetry visual field tests (Humphrey SITA Standard 24-2), excluding tests without reliable indices. VFI progression rate was analysed. VFI progression during the first half period of follow up was compared with that during the second half period. VFI progression was confronted with script alert messages delivered by the last GPA (Guided Progression Analysis) event analysis program. RESULTS: VFI values were "stable or with low progression" in 100% of OHT patients, 88% of early POAG, 38.5% of moderate POAG, 33% of advanced POAG. Progression during the first half period (mean of 3 years) of follow-up could be extrapolated for the second half period in 97% of OHT patients, 76% of early POAG, 70% of moderate POAG, 75% of advanced POAG. Results from VFI trend analysis and GPA event analysis corresponded in 97% of OHT patients, 85% of early POAG, 85% of moderate POAG, and 87% of advanced POAG. DISCUSSION: VFI seems to be a useful indicator for glaucoma evaluation and progression follow-up. It completes the event analysis. Some very advanced POAG cannot be analysed by GPA event analysis. VFI should be able to be used for further follow-up. All these results require validation in larger population. The purpose would be to assess if VFI is able to detect different profiles of progression to help treatment decisions. PMID- 20005006 TI - [Preventing occupational eye trauma (Geneva, Switzerland)]. AB - INTRODUCTION: Occupational eye trauma causes injuries with often serious socioprofessional, medical-legal, and economic consequences, not only for workers themselves, but also for employers. In spite of today's legislation and the efforts to encourage prevention, the frequency of occupational ocular accidents remains relatively high despite how easy it is to protect the eyes. In this study, the reasons that these accidents persist despite the progress made in preventive measures was investigated. MATERIAL AND METHODS: From January to July 2005, we analyzed the parameters related to 175 occupational eye injuries. All patients agreed to take part in this study, which was carried out in the emergency unit of the Ophthalmology Clinic at Geneva University Hospital. Additional data was collected in companies. RESULTS: Construction workers were the most exposed (41.1%). The activity with the greatest risk was grinding (19.4%). The most affected structure of the eye was the cornea (84%), 72.6% patients were not equipped with ocular protection at the time of the accident, and 17.4% wore poorly adapted eye protection. This can be explained by negligence, lack of awareness, etc. Overall, workers, employers, and the legislation in force are all responsible. DISCUSSION: Our results are comparable with those found in the literature, with certain particularities because heavy industry was underrepresented in our sample. Analysis of the law on the prevention of occupational accidents and diseases shows that the worker is not sufficiently aware of his responsibilities. To our knowledge, the legal aspects treated herein have not been studied. As done in certain studies, we emphasize the importance of preventive ophthalmologic examinations as well as improvement of both working conditions and worker awareness in the workplace. RECOMMENDATIONS AND CONCLUSIONS: Primary prevention must be reinforced. Information campaigns within the workplace aimed at workers and revision of the laws on occupational safety are some of the recommendations that are proposed to control occupational ocular accidents. PMID- 20005007 TI - Efficacy of imipenem therapy for Nocardia actinomycetomas refractory to sulfonamides. AB - BACKGROUND: Actinomycetomas are chronic, granulomatous, subcutaneous infections caused by actinomycetes bacteria. Despite prolonged high-dose and combination antibiotic therapies, some cases remain resistant with risks of bone and visceral involvement. OBJECTIVES: We sought to evaluate the efficacy and safety of imipenem monotherapy, and in combination with amikacin for the treatment of severe and refractory disease, and to identify the disease characteristics that might predict therapy failure with first-line sulfonamides. METHODS: A retrospective study was performed of all microbiologically confirmed cases of actinomycetomas treated since 1995 at a tertiary center for mycology. Eleven patients (Nocardia, n = 10) were treated with sulfonamide combinations (trimethoprim/sulfamethoxazole and dapsone). Eight patients (Nocardia, n = 7) refractory to previous therapies including sulfonamides received a 3-week course of either parenteral imipenem monotherapy (1.5 g daily, n = 3) or combination therapy with amikacin (1 g daily, n = 5), which was repeated at 6-month intervals. RESULTS: Eleven patients with limited disease and mean disease duration of 1.7 years responded successfully to sulfonamides after a mean treatment period of 15 months (range 6-48 months). Patients receiving imipenem had mean disease duration of 10 years, with visceral and bone involvement in 4 patients. Imipenem treatment was well tolerated, and 4 patients achieved clinical and microbiological cure after one to two courses of treatment, the others demonstrating greater than 75% clinical improvement and negative culture results. LIMITATIONS: Patient cohorts in this study were small because strict criteria for inclusion included species identification and adequate follow-up periods. The efficacy data for imipenem +/- amikacin therapy cannot be extrapolated to all Nocardia mycetomas, as the cohort treated in this study had particularly refractory infection. CONCLUSIONS: Sulfonamides are effective for limited disease of relatively short duration. Imipenem monotherapy or in combination with amikacin is well tolerated and demonstrates efficacy in severe disease refractory to sulfonamides. PMID- 20005008 TI - Cheilitis glandularis: a clinicopathological study in 22 patients. AB - BACKGROUND: Cheilitis glandularis (CG) is a condition in which thick saliva is secreted by minor labial salivary glands and adheres to a swollen lip causing discomfort to the patient. Most publications refer to single case reports or small case series. OBJECTIVE: We sought to report and to analyze clinical, pathological, and therapeutic data on 22 patients with CG seen at the department of dermatology at our university. METHOD: Retrospective data about 22 patients with CG are reviewed and presented. RESULTS: Seventeen patients were male and 5 were female. All were fair skinned, including 6 albino individuals. Several of them presented significant signs of photodamage on the lips. Surgical treatment was performed in 10 severely affected patients and consisted of a vermilionectomy followed by minor salivary gland removal. Histopathological study revealed various degrees of chronic sialadenitis and vermilion epithelial changes. Superficially invasive and in situ squamous cell carcinoma of the vermilion was detected in 3 cases. LIMITATIONS: Biopsy and surgery were not performed in all patients. CONCLUSIONS: CG is strongly related to sun sensitivity and may be more severe in albino patients. The swollen, sun-exposed lip may become more susceptible to the occurrence of squamous cell carcinoma. PMID- 20005009 TI - Standards for genital protection in phototherapy units. AB - BACKGROUND: Phototherapy is a useful therapy for many dermatologic disorders and is known for its low side-effect profile. However, one potential notable side effect is genital skin cancer. Unfortunately, no standards for genital protection currently exist for this preventable complication. Patients treated with phototherapy may already have a decreased quality of life because of their primary dermatologic disorder. Development of squamous cell carcinoma of the genitalia may certainly further affect the quality of life. OBJECTIVE: The objective was to determine which readily available materials afford the best photoprotection of the male genitalia. METHODS: Seven common materials used in phototherapy units for genital protection were placed over ultraviolet (UV) B and UVA monitors and placed in broadband UVB, narrowband UVB, and UVA full-body units. The percentage of light blocked was then calculated. RESULTS: Blue and white cotton underwear, blue surgical towels, an athletic supporter with or without a cup, and the psoralen plus UVA pouch provided acceptable means of genital protection; however, surgical masks did not. LIMITATIONS: Only the most commonly used materials were tested in the phototherapy units. The materials were not of a single material type or similar masses. In addition, only one of each type of full-body phototherapy unit was used to obtain the data. CONCLUSION: Although a polyester composition provides better UV protection, factors such as low porosity and higher mass are intrinsic to decreasing the amount of UV penetration of any fabric. Of the commonly used objects, surgical masks do not provide sufficient protection to the genital area. PMID- 20005010 TI - Do emergency physicians attribute drug-related emergency department visits to medication-related problems? AB - STUDY OBJECTIVE: Adverse drug events represent the most common cause of preventable nonsurgical adverse events in medicine but may remain undetected. Our objective is to determine the proportion of drug-related visits emergency physicians attribute to medication-related problems. METHODS: This prospective observational study enrolled adults presenting to a tertiary care emergency department (ED) during 12 weeks. Drug-related visits were defined as ED visits caused by adverse drug events. The definition of adverse drug event was varied to examine both narrow and broad adverse drug event classification systems. Clinical pharmacists evaluated all patients for drug-related visits, using standardized assessment algorithms, and then followed patients until hospital discharge. Interrater agreement for the clinical pharmacist diagnosis of drug-related visit was assessed. Emergency physicians, blinded to the clinical pharmacist opinion, were interviewed at the end of each shift to determine whether they attributed the visit to a medication-related problem. An independent committee reviewed and adjudicated all cases in which the emergency physicians' and clinical pharmacists' assessments were discordant, or either the emergency physician or clinical pharmacist was uncertain. The primary outcome was the proportion of drug related visits attributed to a medication-related problem by emergency physicians. RESULTS: Nine hundred forty-four patients were enrolled, of whom 44 patients received a diagnosis of the narrowest definition of an adverse drug event, an adverse drug reaction (4.7%; 95% confidence interval [CI] 3.5% to 6.2%). Twenty-seven of these were categorized as medication-related by emergency physicians (61.4%; 95% CI 46.5% to 74.3%), 10 were categorized as uncertain (22.7%; 95% CI 12.9% to 37.1%), and 7 categorized as a non-medication-related problem (15.9%; 95% CI 8.0% to 29.5%). Seventy-eight patients (8.3%; 95% CI 6.7% to 10.2%) received a diagnosis of an adverse drug event caused by an adverse drug reaction, a drug interaction, drug withdrawal, a medication error, or noncompliance. Emergency physicians attributed 49 of these to a medication related problem (62.8%; 95% CI 51.7% to 72.7%), were uncertain about 15 (19.2%; 95% CI 12.0% to 29.4%), and attributed 14 to non-medication-related problems (17.9%; 95% CI 11.0% to 27.9%). Twenty-five of 29 (86.2%; 95% CI 69.3% to 94.4%) adverse drug events not considered medication related by emergency physicians were rated at least moderate in severity. CONCLUSION: A significant proportion of drug-related visits are not deemed medication related by emergency physicians. Drug-related visits not attributed to medication-related problems by emergency physicians may be missed in ongoing outpatient adverse drug event surveillance programs intended to develop strategies to enhance drug safety. Further research is needed to determine what the effect may be of not attributing adverse drug events to medication-related problems. PMID- 20005011 TI - Medication errors recovered by emergency department pharmacists. AB - STUDY OBJECTIVE: We assess the impact of emergency department (ED) pharmacists on reducing potentially harmful medication errors. METHODS: We conducted this observational study in 4 academic EDs. Trained pharmacy residents observed a convenience sample of ED pharmacists' activities. The primary outcome was medication errors recovered by pharmacists, including errors intercepted before reaching the patient (near miss or potential adverse drug event), caught after reaching the patient but before causing harm (mitigated adverse drug event), or caught after some harm but before further or worsening harm (ameliorated adverse drug event). Pairs of physician and pharmacist reviewers confirmed recovered medication errors and assessed their potential for harm. Observers were unblinded and clinical outcomes were not evaluated. RESULTS: We conducted 226 observation sessions spanning 787 hours and observed pharmacists reviewing 17,320 medications ordered or administered to 6,471 patients. We identified 504 recovered medication errors, or 7.8 per 100 patients and 2.9 per 100 medications. Most of the recovered medication errors were intercepted potential adverse drug events (90.3%), with fewer mitigated adverse drug events (3.9%) and ameliorated adverse drug events (0.2%). The potential severities of the recovered errors were most often serious (47.8%) or significant (36.2%). The most common medication classes associated with recovered medication errors were antimicrobial agents (32.1%), central nervous system agents (16.2%), and anticoagulant and thrombolytic agents (14.1%). The most common error types were dosing errors, drug omission, and wrong frequency errors. CONCLUSION: ED pharmacists can identify and prevent potentially harmful medication errors. Controlled trials are necessary to determine the net costs and benefits of ED pharmacist staffing on safety, quality, and costs, especially important considerations for smaller EDs and pharmacy departments. PMID- 20005012 TI - Neurostructural predictors of Alzheimer's disease: a meta-analysis of VBM studies. AB - The identification of biological markers at early stages of Alzheimer's disease (AD) contributes to diagnostic accuracy and adds prognostic value. However, in spite of recent developments, results of neurostructural imaging studies on predicting conversion to AD are not uniform. We conducted a systematic review of voxel-based morphometry (VBM) studies about the neurostructural predictors of conversion to AD. Ten studies met inclusion criteria and nine reported baseline regional gray matter (GM) atrophy in mild cognitive impairment (MCI) or healthy subjects who progressed to AD. Using the method of Activation Likelihood Estimation, we meta-analyzed the coordinates from the six longitudinal VBM studies that enrolled subjects with amnestic MCI (aMCI) at baseline. These comprised a total of 429 aMCI subjects, of which 142 converted to AD. Meta analysis yielded one significant cluster of GM volumetric reduction in aMCI patients who converted to AD, located in the left hippocampus and parahippocampal gyrus. In conclusion, left medial temporal lobe atrophy is the most consistent neurostructural biomarker to predict conversion from aMCI to AD. PMID- 20005013 TI - [Intermittent obstructive jaundice by Lemmel's syndrome]. PMID- 20005014 TI - [Mesalazine-induced acute pericarditis]. PMID- 20005015 TI - [Proton pump inhibitors and clopidogrel in a patient with cardiovascular risk factors: Cardiovascular versus gastrointestinal risk?]. PMID- 20005016 TI - [Risk factors and early diagnosis of pancreatic cancer]. AB - Pancreatic cancer (PC) is usually incurable. Identifying people at risk for developing PC may improve the prognosis of this entity. The main risk factors for PC are Peutz-Jeghers syndrome, hereditary pancreatitis and a history family of PC. Other factors, such as advanced age and smoking, should also be taken into account. PC screening is only useful in very high risk individuals. Tools that allow tumors to be identified in the early stages are required in order to apply appropriate curative treatments. In this scenario, only endoscopic ultrasound with cytological analysis of suspicious pancreatic lesions has proved to be useful. PMID- 20005017 TI - [Family and Community Medicine Training Program: Theory and practice]. AB - OBJECTIVE: To improve and adapt training rotation periods following the training program (POE) of the Badajoz Family and Community Medicine (MFyC) Training Unit (UDOCBA). DESIGN: Mixed qualitative (brainstorming and nominal technical group), and quantitative (cross-sectional descriptive), study in 2006 and 2007. SETTING: UDOCBA. PARTICIPANTS: The participants include 50 Residents, 31 accredited tutors, a health technician and a UDOCBA coordinator, as well as 93 MFyC Teaching Units in Spain (UUDD). INTERVENTIONS AND MEASUREMENTS: A rotations working group was formed in UDOCBA, which met on two occasions to make substantial modifications to adapt the POE. The first meeting was a brainstorming session, and the second using a nominal technical group, whose results were submitted to an Advisory Committee. MATERIALS: Questionnaire aimed at tutors, analysis of residents self-assessment files, a questionnaire delivered to all the UUDD (via e mail) and nominal resident groups. Descriptive and bivariate statistics on the variables using the SPSS 15 program. RESULTS: The consensus of the second meeting of the UDOCBA rotations group on evaluating the results obtained with the rest of the tools used, were: to shorten the initial rotation of the C.S. to 5 months, increase the cardiology rotation to 3 months, shorten the rural rotation to 2 months and opt out of the elective 1 month stay in palliatives and radiology. CONCLUSIONS: Theoretically well prepared and easily to put into practice. All the consensus results of the UDOCBA were accepted by the Advisory Committee, followed by POE directives. Homogeneity was seen in the UUDD rotations. Unanimity of initial and final rotation by the C.S., and less in rural. PMID- 20005018 TI - Captopril renography as a prognostic factor in obstructive hydronephrosis with preserved renal function. AB - Hydronephrotic kidney with a differential renal function greater than 55% is defined as supranormal. The signification of this finding remains controversial. In this article, the authors reported a case of supranormal function in obstructive hydronephrosis. Differential renal functions were evaluated after administration of captopril and after pyeloplasty. The role of captopril renography as a prognostic factor for surgery is discussed. PMID- 20005020 TI - Antimicrobial study of newly synthesized 6-substituted indolo[1,2-c]quinazolines. AB - A new series of indolo[1,2-c]quinazoline derivatives were prepared in good yield through reaction of 2-(o-aminophenyl)indole with a variety of arylaldehydes. The structures of the newly synthesized compounds were confirmed by IR, (1)H NMR, (13)C NMR and mass spectral studies and elemental analysis. All the title compounds were investigated for their activity against certain strains of Gram positive bacteria (Staphylococcus aureus, Bacillus subtilis and Streptococcus pyogenes), Gram-negative bacteria (Salmonella typhimurium, Escherichia coli and Klebsiella pneumonia) and pathogenic Fungi (Aspergillus niger, Candida albicans and Trichoderma viridae). Ampicillin and ketoconazole were used as reference compounds. The results revealed that some of synthesized compounds displayed marked activity against all the tested microorganisms. PMID- 20005019 TI - [On the way to shortening tuberculosis treatments: clinical trials of the Unitat d' Investagacio en Tuberculosi de Barcelona supported by the Centers for Disease Control and Prevention]. AB - New treatment guidelines are required to improve the tuberculosis control strategies that have been used for 30 years. Seven centers of the Barcelona Tuberculosis Research Unit (BTRU) (Unitat d'Investigacio en Tuberculosi de Barcelona) are collaborating with the Division of Tuberculosis Elimination of the United States Centers for Disease Control and Prevention in a series of clinical trials on latent tuberculosis infection and tuberculosis disease. BTRU participation began in 2004 with Study 26, an evaluation of the efficacy and tolerability of rifapentine plus isoniazid administered once weekly for 3 months compared with the standard treatment for latent tuberculosis infection. The BTRU centers together enrolled 246 patients (3% of the total). General enrollment was completed in February, 2008. HIV-infected patient and child enrollment continues. Treatment with 12 doses instead of 270 doses is expected to be a clear success. However, the analysis will be completed in 2010. Study 28 (started in 2006), designed for the treatment of pulmonary tuberculosis, compared standard treatment with an experimental regimen substituting moxifloxacin for isoniazid. BTRU centers together enrolled 15 patients (3.5% of the total). The provisional results (presented at the 47th Interscience Conference on Antimicrobial Agents and Chemotherapy in Chicago, 2007) showed no difference between the sputum conversion rate of each regimen at week 8 of treatment. Study 29 is currently underway, in which rifapentine was introduced in the experimental regimen for active tuberculosis treatment. PMID- 20005021 TI - Synthesis of glycoside derivatives of hydroxyanthraquinone with ability to dissolve and inhibit formation of crystals of calcium oxalate. Potential compounds in kidney stone therapy. AB - Synthesis of glycosyl derivatives of hydroxyanthraquinones (6-10) potentially useful for kidney stone therapy is presented. These compounds were analyzed as inhibitors of calcium oxalate crystals formation as well as substances with the ability of dissolving crystalline calcium oxalate. In addition, the effect of the compounds obtained on real kidney stones was analyzed by ex vivo tests. The tests on L929 and A545 cell lines have shown that the compounds obtained were not cytotoxic. PMID- 20005022 TI - Identification of antibacterial and antifungal pharmacophore sites for potent bacteria and fungi inhibition: indolenyl sulfonamide derivatives. AB - Synthesis of seven new indolenyl sulfonamides, have been prepared by the condensation reaction of indole-3-carboxaldehyde with different sulfonamides such as, sulphanilamide, sulfaguanidine, sulfathiazole, sulfamethoxazole, sulfisoxazole, sulfadiazine and sulfamethazine. These synthesized compounds have been used as potential ligands for complexation with some selective divalent transition metal ions (cobalt, copper, nickel & zinc). Structure of the synthesized ligands has been deduced from their physical, analytical (elemental analyses) and spectral (IR, (1)H NMR and (13)C NMR & UV-vis) data. All the compounds have also been assayed for their in vitro antibacterial and antifungal activities examining six species of pathogenic bacteria (Escherichia coli, Shigella flexneri, Pseudomonas aeruginosa, Salmonella typhi, Staphylococcus aureus and Bacillus subtilis) and six of fungi (Trichophyton longifusus, Candida albicans, Aspergillus flavus, Microsporum canis, Fusarium soloni and Candida glabrata). Antibacterial and antifungal results showed that all the compounds showed significant antibacterial activity whereas most of the compounds displayed good antifungal activity. Brine shrimp bioassay was also carried out for in vitro cytotoxic properties against Artemia salina. PMID- 20005023 TI - [Recurrent hypoglycemia in a 68-year-old man]. PMID- 20005024 TI - Attachment, belonging and identity are important to effective health curricula. PMID- 20005025 TI - Measurement of gross alpha and gross beta activity concentrations in human tooth. AB - The gross alpha and gross beta activity concentrations were measured in human tooth taken from 3 to 6 age-groups to 40 and over ones. Accumulated teeth samples are investigated in two groups as under and above 18 years. The gross alpha and beta radioactivity of human tooth samples was measured by using a gas-flow proportional counter (PIC-MPC 9604-alpha/beta counter). In tooth samples, for female age-groups, the obtained results show that the mean gross alpha and gross beta activity concentrations varied between 0.534-0.203 and 0.010-0.453 Bq g(-1) and the same concentrations for male age-groups varied between 0.009-1.168 and 0.071-0.204 Bq g(-1), respectively. PMID- 20005026 TI - Injury to the brachial plexus in neck dissections. AB - Injury to the brachial plexus is a rare complication of neck dissection. We present a case of a 65-year-old man who developed a lesion of the nerve root of C5 and C6 as a result of radical neck dissection. PMID- 20005027 TI - Measuring the best outcome for the least intervention: can the Optimality Index US be applied in the UK? AB - OBJECTIVE: to pilot the Optimality Index-US (OI-US) for the first time within a UK maternity setting in a sample of women at mixed risk. DESIGN: a multidisciplinary group reviewed the items and evidence base of the OI-US. A pilot study was undertaken to compare the availability and quality of data from maternity records to complete the OI-US. Data were collected from maternity records. SETTING: a maternity unit of an inner city teaching hospital in England. PARTICIPANTS: clinical midwives, research midwives, midwifery lecturers and consultant obstetricians (n=10) reviewed the items and evidence base of the OI US. Data were collected from the maternity records of 97 women receiving caseload care and 103 women receiving standard care. MEASUREMENTS AND FINDINGS: when the multidisciplinary group reviewed the items and evidence base of the OI-US, it was noted that some social and clinical factors should be considered for inclusion as part of the Perinatal Background Index (PBI) and OI. The results suggest that the inclusion of women at higher risk in this sample within the UK maternity setting has not been captured by the OI-US. KEY CONCLUSIONS: the following social and clinical factors should be included as part of the PBI and OI for the UK setting: measure of social deprivation, woman's ability to speak and understand English in relation to accessing maternity care, mental health problems during pregnancy and history of domestic violence during pregnancy IMPLICATIONS FOR PRACTICE: availability of items in electronic records is poor and it is recommended that the OI-UK version is a useful research tool in prospective data collection. The development of an international version would be valuable for comparison of background risk and outcomes across a range of care settings. PMID- 20005028 TI - The effect of obesity and gender on body segment parameters in older adults. AB - BACKGROUND: Anthropometry is a necessary aspect of aging-related research, especially in biomechanics and injury prevention. Little information is available on inertial parameters in the geriatric population that account for gender and obesity effects. The goal of this study was to report body segment parameters in adults aged 65 years and older, and to investigate the impact of aging, gender and obesity. METHODS: Eighty-three healthy old (65-75 years) and elderly (>75 years) adults were recruited to represent a range of body types. Participants underwent a whole body dual energy X-ray absorptiometry scan. Analysis was limited to segment mass, length, longitudinal center of mass position, and frontal plane radius of gyration. A mixed-linear regression model was performed using gender, obesity, age group and two-way and three-way interactions (alpha=0.05). FINDINGS: Mass distribution varied with obesity and gender. Males had greater trunk and upper extremity mass while females had a higher lower extremity mass. In general, obese elderly adults had significantly greater trunk segment mass with less thigh and shank segment mass than all others. Gender and obesity effects were found in center of mass and radius of gyration. Non-obese individuals possessed a more distal thigh and shank center of mass than obese. Interestingly, females had more distal trunk center of mass than males. INTERPRETATION: Age, obesity and gender have a significant impact on segment mass, center of mass and radius of gyration in old and elderly adults. This study underlines the need to consider age, obesity and gender when utilizing anthropometric data sets. PMID- 20005029 TI - Prevention of diabetic kidney disease: negative clinical trials with renin angiotensin system inhibitors. PMID- 20005030 TI - Pathophysiology and management of preeclampsia-associated severe hyponatremia. PMID- 20005031 TI - Impacts of changes in land use and fragmentation patterns on Atlantic coastal forests in northern Spain. AB - Changes in forested landscapes may have important consequences for ecosystem services and biodiversity conservation. In northern Spain, major changes in land use occurred during the second half of the 20th century, but their impacts on forests have not been quantified. We evaluated the dynamics of landscape and forest distribution patterns between 1957 and 2003 in Fragas do Eume Natural Park (northwestern Spain). We used orthoimages and a set of standard landscape metrics to determine transitions between land cover classes and to examine forest distribution patterns. Eucalypt plantations showed the greatest increase in area (197%) over time. Furthermore, transitions to eucalypt plantations were found in all major land cover classes. Forest showed a net decline of 20% in total area and represented 30% of the landscape area in 2003. Forest losses were mainly due to eucalypt plantations and the building of a water reservoir, while forest gains were due to increases in shrubland, meadows and cultivated fields which had been recolonised. Forest patch size and core area decreased, and edge length increased over time. In turn, increases were obtained in mean distance between forest patches, and in adjacency to eucalypt plantations and to a water reservoir. These results suggest an increase in forest fragmentation from 1957 to 2003, as well as a change in the nature of the habitat surrounding forest patches. This study shows that land use changes, mostly from eucalypt plantation intensification, negatively affected forested habitats, although some regeneration was ongoing through ecological succession from land abandonment. PMID- 20005032 TI - Rare E196K mutation in the PRNP gene of a patient exhibiting behavioral abnormalities. AB - Genetic transmissible spongiform encephalopathies (TSEs) account for approximately 10-15% of overall human prion diseases worldwide, but genotype phenotype correlations remain incomplete. Here we report the case of an 80-year old man who developed rapidly progressive behavioral abnormalities and myoclonus following a stroke. Repeated electroencephalography (EEG) revealed a general slowing of the basic activity, as well as several episodes of triphasic waves, with neither periodic activity nor recorded seizure. 14.3.3 protein was detected in cerebral cerebrospinal fluid, and direct sequencing of the PRNP gene showed an E196K mutation associated with homozygosity for methionine at codon 129. The patient was diagnosed with probable genetic prion disease with a Creutzfeldt Jakob disease-like phenotype. The PRNP E196K mutation has only rarely been described in the literature, and generally patients exhibited an atypical initial phenotype, mainly involving abnormal behavioral features. Further observations are needed to confirm this particular clinical pattern associated with the mutation. PMID- 20005033 TI - Predictive factors for outcome of invasive video-EEG monitoring and subsequent resective surgery in patients with refractory epilepsy. AB - OBJECTIVE: This is a descriptive study of patients who underwent invasive video EEG monitoring (IVEM) at Ghent University Hospital. The aim of the study is to identify predictive factors for outcome of IVEM and resective surgery (RS). These factors may optimize the patient flow following the non-invasive presurgical evaluation towards IVEM and RS or other treatments. PATIENTS AND METHODS: Over the past 16 years, 68/710 refractory epilepsy patients included in the presurgical evaluation protocol (M/F 41/27, mean age 33 years) underwent IVEM at Ghent University Hospital. Patient features and follow-up data were collected from the patients' medical files and the electronic patient database at the neurology and neurosurgery department. Predictive factors for IVEM outcome were identified by comparing features of patients with a positive IVEM outcome (i.e. ictal onset zone identification) and patients with a negative IVEM outcome. Predictive factors for RS outcome were identified by comparing features of patients with Engel class I and patients with Engel class II-IV outcome. RESULTS: In 56/68 patients (82%) IVEM outcome was positive. The occurrence of a seizure free interval in the patient's history and a non-localizing ictal scalp EEG in patients with a structural abnormality on MRI (p<0.05) were predictive factors for a negative IVEM outcome. 32/68 patients underwent RS. In 22/32 (70%) patients RS resulted in an Engel class I outcome. A structural abnormality on MRI was a predictive factor for a positive RS outcome in patients in whom a focal or regional focus was resected (p<0.05). CONCLUSION: This study shows that IVEM identifies one or more ictal onset zone(s) in up to 80% of patients. The potential of IVEM to identify the ictal onset zone is unlikely in patients with a seizure-free interval in their medical history and a non-localizing ictal scalp EEG during the non-invasive presurgical evaluation. Half of these patients underwent RS with long-term seizure freedom in 70%. Patients with structural MRI lesions have the highest chance of seizure freedom. These findings may contribute to the optimization of patient management during both the invasive and non invasive presurgical work-up. PMID- 20005034 TI - Toxicity of purified terephthalic acid manufacturing wastewater on reproductive system of male mice (Mus musculus). AB - Reproductive toxicity of purified terephthalic acid (PTA) manufacturing wastewater on the male mice (Mus musculus) was investigated after 35-day intragastric perfusion treatment with the wastewater. Fluorescein diacetate and propidium iodide staining, and flow cytometry were used to assess the toxicity of PTA wastewater on spermatogenic cells. PTA wastewater induced significant variations in the relative percentages of immature haploid, diploid, tetraploid and S-phase spermatogonia. Percentage of viable spermatogenic cells was reduced from 93.1+/-2.3 in control group to 90.4+/-1.9 in the wastewater-treated group. Testicular histopathology revealed expansion of interstitial space and reduction in the number and size of Leydig cells induced by the wastewater, which was further certified by the decrease (10.6%) in relative testes weight and the increase (101.3%) in sperm shape abnormality in the wastewater-treated group. In this study, PTA wastewater was found to have reproductive toxicity on male mice, and public health problems may potentially arise from the discharge of the wastewater into the environment. PMID- 20005035 TI - Photodecolorization of Eriochrome Black T using NiS-P zeolite as a heterogeneous catalyst. AB - NiS-P zeolite was prepared by ion exchange and precipitation procedures and it was characterized by FT-IR, SEM and thermal methods. The prepared composite was used as a catalyst in the photodecolorization process of Eriochrome Black T (E.B.T.) dye in aqueous solution under UV irradiation. The effect of key operating parameters such as catalyst dosage, temperature, initial concentration of the dye and initial pH of the solutions were studied on the decolorization process of dye. The primary objective was to determine the optimal conditions for each of the parameters. UV-vis spectrophotometric measurements were performed for the determination of decolorization and mineralization extents. The optimal operation parameters were found as follows: pH 9.1, 0.8 g L(-1) of catalyst loading and 40 ppm of dye concentration. The NiS particles out of zeolite framework did not show significant decolorization efficiency. The decolorization process obeyed first-order kinetics. PMID- 20005036 TI - A mechanistic kinetic model for phenol degradation by the Fenton process. AB - The objective of this paper is to develop and validate a mechanistic model for the degradation of phenol by the Fenton process. Experiments were performed in semi-batch operation, in which phenol, catechol and hydroquinone concentrations were measured. Using the methodology described in Pontes and Pinto [R.F.F. Pontes, J.M. Pinto, Analysis of integrated kinetic and flow models for anaerobic digesters, Chemical Engineering Journal 122 (1-2) (2006) 65-80], a stoichiometric model was first developed, with 53 reactions and 26 compounds, followed by the corresponding kinetic model. Sensitivity analysis was performed to determine the most influential kinetic parameters of the model that were estimated with the obtained experimental results. The adjusted model was used to analyze the impact of the initial concentration and flow rate of reactants on the efficiency of the Fenton process to degrade phenol. Moreover, the model was applied to evaluate the treatment cost of wastewater contaminated with phenol in order to meet environmental standards. PMID- 20005037 TI - Design of an electro-Fenton system with a novel sandwich film cathode for wastewater treatment. AB - In this study, we demonstrate an electro-Fenton (E-Fenton) system constructed with a novel sandwich film cathode (SFC). For the fabrication of SFC, Fe(2+) chitosan (Fe-CHI) was first deposited on foam nickel (Fe-CHI/Ni). Then two pieces of Fe-CHI/Ni was used to fasten one piece of activated carbon fiber (ACF) to obtain a Fe-CHI/Ni|ACF|Fe-CHI/Ni sandwich film cathode. We interestingly found that this SFC based E-Fenton system could effectively degrade rodamine B with in situ generating both hydrogen peroxide and iron ions. Its degradation efficiency was significantly higher than those of the E-Fenton systems constructed with composite cathodes of carbon nanotubes with Fe@Fe(2)O(3) core-shell nanowires or Cu(2)O nanocubes reported in our previous studies. Hydrogen peroxide electrogenerated through the reduction of O(2) adsorbed on the sandwich film cathode and the iron ions produced by the leakage from Fe(2+)-chitosan film during the E-Fenton reaction were, respectively, monitored, providing clues to understand the high efficiency of this novel SFC based E-Fenton system. More importantly, this low-cost sandwich film cathode was very stable and could be reused without catalytic activity decrease, suggesting its potential application in the wastewater treatment. PMID- 20005038 TI - Trajectory measurements of a wall jet impinging onto a forward facing step entering a cross-flow. AB - This study examines a horizontal wall jet impinging onto a forward facing step in a cross-flow. Planar laser induced fluorescence (PLIF) experiments in a water channel indicate that the wall-jet flow after impinging onto the step, becomes a vertical jet with an elliptical cross section. Experiments indicate that the jet trajectory scales with the perimeter of the elliptical jet issuing vertically into the cross-flow. The trajectory consists of three regions: the near-field region which is well described by a power law with an exponent of 1/2, the mid field region where the jet is fully bent over which is described by a power law with an exponent of 1/3, and a far-field region where the jet is dominated by the cross-flow. This paper provides a prediction of the plume behaviour based on the geometric and initial conditions of the jet (diameter, step height, distance from jet to step, and velocity ratio) alone. The Briggs entrainment model for a round jet was also used to predict the trajectories of the jet in the cross-flow. It was found that the entrainment coefficients, alpha and beta, for the elliptical jet case had average values of 0.15 and 0.58 respectively. PMID- 20005039 TI - Incorporation of mineral phosphorus and potassium on leather waste (collagen): a new N(collagen)PK-fertilizer with slow liberation. AB - The feasibility of using a solid waste (rich in nitrogen) from the leather industry, after chromium extraction, as adsorbent for P and K, for possible utilization as NPK fertilizer was evaluated. The materials, with and without the addition of P and K, were characterized by chemical analyses, infrared spectroscopy, EDS (energy dispersive X-ray spectrometry) and SEM (scanning electronic microscopy). Langmuir and Freundlich equations were used for analyzing the experimental data, which showed a better fit to the Freundlich model, thus suggesting a multilayer adsorption process on the surface of the adsorbent. A preliminary test in greenhouse demonstrates that the P and K incorporation on the matrix rich in nitrogen (collagen) is a interesting alternative to use such material as NPK fertilizer. The application of N(collagen)PK formulations, as a source of nutrients for the growth of rice plants, showed promising agronomic results. PMID- 20005040 TI - Spinal macrophage migration inhibitory factor contributes to the pathogenesis of inflammatory hyperalgesia in rats. AB - Pro-inflammatory cytokine production after nociceptive stimuli is pivotal for hyperalgesia. As macrophage migration inhibitory factor (MIF), a pleiotropic cytokine produced mainly by nonneuronal tissue, has been involved in the regulation of neuronal functions, herein we examined the role for MIF in formalin induced inflammatory pain model. MIF critically contributed to nociceptive behaviors following formalin injection. Specifically, spinal administration of a MIF inhibitor (ISO-1) prevented and reversed flinching responses in rats. Further examination showed that levels of both MIF and the MIF receptor CD74 were substantially increased within the ipsilateral spinal cord dorsal horn after formalin administration. Mechanistic studies revealed that MIF upregulated the expression of the spinal NMDA receptor subunit NR2B via the MAPK signaling pathway. Moreover, microglial cells were found to be the major source of spinal MIF after formalin administration by fluorescence colocalization. These data highlight spinal MIF plays a critical role in the pathogenesis of formalin induced inflammatory pain and suggest MIF may be a potential target for therapy of such pathological condition. PMID- 20005042 TI - Pain sensation evoked by observing injury in others. AB - Observing someone else in pain produces a shared emotional experience that predominantly activates brain areas processing the emotional component of pain. Occasionally, however, sensory areas are also activated and there are anecdotal reports of people sharing both the somatic and emotional components of someone else's pain. Here we presented a series of images or short clips depicting noxious events to a large group of normal controls. Approximately one-third of this sample reported an actual noxious somatic experience in response to one or more of the images or clips. Ten of these pain responders were subsequently recruited and matched with 10 non-responders to take part in an fMRI study. The subjects were scanned while observing static images of noxious events. In contrast with emotional images not containing noxious events the responders activated emotional and sensory brain regions associated with pain while the non responders activated very little. These findings provide convincing evidence that some people can readily experience both the emotional and sensory components of pain during observation of other's pain resulting in a shared physical pain experience. PMID- 20005043 TI - Exploring the brain in pain: activations, deactivations and their relation. AB - The majority of neuroimaging studies on pain focuses on the study of BOLD activations, and more rarely on deactivations. In this study, in a relatively large cohort of subjects (N=61), we assess (a) the extent of brain activation and deactivation during the application of two different heat pain levels (HIGH and LOW) and (b) the relations between these two directions of fMRI signal change. Furthermore, in a subset of our subjects (N=12), we assess (c) the functional connectivity of pain-activated or -deactivated regions during resting states. As previously observed, we find that pain stimuli induce intensity dependent (HIGH pain>LOW pain) fMRI signal increases across the pain matrix. Simultaneously, the noxious stimuli induce activity decreases in several brain regions, including some of the 'core structures' of the default network (DMN). In contrast to what we observe with the signal increases, the extent of deactivations is greater for LOW than HIGH pain stimuli. The functional dissociation between activated and deactivated networks is further supported by correlational and functional connectivity analyses. Our results illustrate the absence of a linear relationship between pain activations and deactivations, and therefore suggest that these brain signal changes underlie different aspects of the pain experience. PMID- 20005045 TI - Analysis of Kikuchi band contrast reversal in electron backscatter diffraction patterns of silicon. AB - We analyze the contrast reversal of Kikuchi bands that can be seen in electron backscatter diffraction (EBSD) patterns under specific experimental conditions. The observed effect can be reproduced using dynamical electron diffraction calculations. Two crucial contributions are identified to be at work: First, the incident beam creates a depth distribution of incoherently backscattered electrons which depends on the incidence angle of the beam. Second, the localized inelastic scattering in the outgoing path leads to pronounced anomalous absorption effects for electrons at grazing emission angles, as these electrons have to go through the largest amount of material. We use simple model depth distributions to account for the incident beam effect, and we assume an exit angle dependent effective crystal thickness in the dynamical electron diffraction calculations. Very good agreement is obtained with experimental observations for silicon at 20keV primary beam energy. PMID- 20005044 TI - N-Methyl-D-aspartate receptor (NMDAR) independent maintenance of inflammatory pain. AB - Following peripheral inflammation, NMDA receptor (NMDAR) activation in spinal cord dorsal horn neurons facilitates the generation of pain in response to low threshold inputs (allodynia) and signals the phosphorylation of protein kinase C (pPKC) and extracellular signal-regulated kinase 2 (pERK2). Intraplantar complete Freund's adjuvant (CFA) induces inflammatory nociception (allodynic pain) at 24 hours (h) with a concurrent increase in neuronal pPKCgamma and pERK2 but not glial pERK2. These effects are attenuated in a spatial knockout of the NMDAR (NR1 KO) confined to SCDH neurons. Although glia and proinflammatory cytokines are implicated in the maintenance of inflammatory pain and neuronal activation, the role of NMDARs and neuronal-glial-cytokine interactions that initiate and maintain inflammatory pain are not well defined. In the maintenance phase of inflammatory pain at 96h after CFA the NR1 KO mice are no longer protected from allodynia and the SCDH expression of pPKCgamma and pERK2 are increased. At 96h the expression of the proinflammatory cytokine, IL-1beta, and pERK2 are increased in astrocytes. Intrathecal IL-1 receptor antagonist (IL-1ra), acting on neuronal IL-1 receptors, completely reverses the allodynia at 96h after CFA. Deletion of NMDAR-dependent signaling in neurons protects against early CFA-induced allodynia. Subsequent NMDAR-independent signaling that involves neuronal expression of pPKCgamma and the induction of pERK2 and IL-1beta in activated astrocytes contributes to the emergence of NMDAR-independent inflammatory pain behavior at 96h after CFA. Effective reduction of the initiation and maintenance of inflammatory pain requires targeting the neuron-astrocyte-cytokine interactions revealed in these studies. PMID- 20005046 TI - Effects of diclazuril on apoptosis and mitochondrial transmembrane potential in second-generation merozoites of Eimeria tenella. AB - Diclazuril, a benzeneacetonitrile anticoccidial agent, has potent activity against various stages of Eimeria tenella (E. tenella). To study the effects of diclazuril on E. tenella merozoites, purified second-generation merozoites were obtained from infected chicken caecal tissue at 120h after inoculation by a combination of enzymatic digestion, centrifugation, erythrocytes disruption and percoll density gradient centrifugation. Ultrastructural changes were monitored by transmission electron microscopy (TEM). Apoptosis and mitochondrial transmembrane potential were determined by flow cytometry (FCM). The results showed that diclazuril induced ultrastructural changes and significantly increased the ratio of early apoptosis by 180.75% (P<0.01) and late apoptosis/necrosis by 86.82% (P<0.05) in second-generation merozoites, respectively. Compared with the infected/control group, the ratio of second generation merozoites that lost mitochondrial function was increased by 45.04% (P<0.01) in the infected/treatment group. In conclusion, diclazuril induced morphological changes and attenuated the activity of mitochondrial transmembrane potential of merozoites, which is involved in mitochondrial-depended apoptosis in second-generation merozoites of E. tenella. PMID- 20005048 TI - Burn injuries due to Iranian traditional customs. PMID- 20005049 TI - The treatment of hand burns: timing of debridement and grafting. PMID- 20005051 TI - The significance of incomplete skull fracture in the birth injury. AB - Vaginal delivery is accomplished by the force of the labor overcoming the resistance forces of birth canal. During this process, the fetal head passes through the birth canal and the skull receives pressure on the lateral aspect, resulting in molding, the convex shaping of the cranium. Also, the infant's skull is compressed by the mother's pelvic bony structures. These forces may lead to skull fractures and brain injuries. The hypothesis by the authors is that many skull fractures of the newborn present as incomplete fractures. The bony skull of the newborn is histologically primary bone tissue and which is incomplete in its ossification process. During birth the pressure forces upon the newborn's skull is gradual in one direction, rather than a sudden impact, and therefore it is thought that the skull fracture would be an incomplete fracture. However, it is very hard to ascertain the presence of incomplete fractures especially in incompletely ossified skulls with plain X-ray studies, and therefore it is possible that the real incidence of skull fractures in the newborn are higher than reported in the current and past literature. It is also probable that the external forces upon the skull that are sufficient to cause skull fractures, would also lead to significant brain injury more frequently than actually observed, and subsequently contribute to development of many brain disease later in children. The authors of this study propose that very close examination should be conducted to find incomplete fracture, and increased efforts should be made to establish the presence of possible accompanied brain injuries in babies with incomplete skull fracture. The definitive diagnosis and treatment, as well as close follow up of patients with brain injury will assist the clinician in determining the causes of neurological diseases especially in those with previously unknown etiologies, which may be due to birth injuries. Assistance may be also afforded in the early treatment and prevention of such conditions. PMID- 20005052 TI - Prophylactic topical heparin can prevent or postpone intravenous cannula induced superficial thrombophlebitis. AB - Intravenous cannulation is a cornerstone of today's medical practice. Maintaining a single indwelling intravenous (IV) cannula for long duration is limited by the development of superficial thrombophlebitis (ST). It is a self limiting inflammation and thrombosis of superficial veins. ST presents with fever, pain, erythema, tenderness and cord like swelling. The incidence of ST is high and usually occurs within 72h of IV cannula insertion. The current standard medical therapy for ST is topical heparin application for 7 days. Heparin acts by preventing coagulation rather than lysing a formed clot. So, if topical heparin is started prophylactically even before ST sets in, i.e. from day 1 of IV cannula insertion it can prevent or postpone ST more effectively. It increases the indwelling time of a single IV cannula and can be very useful in high risk groups requiring IV cannulation like patients receiving cancer chemotherapy, ICU patients and infants. It decreases the need for recurrent cannulations and associated morbidity thereby improving patient compliance. It also prevents extended hospital stay due to ST and related complications. High incidence of ST justifies the use of prophylactic topical heparin with all IV cannulations. Prophylaxis will be better than treatment in managing patients with IV cannulas. PMID- 20005050 TI - Prenatal thyroxine treatment disparately affects peripheral and amygdala thyroid hormone levels. AB - A prenatal hypothyroid state is associated with behavioral abnormalities in adulthood. Wistar Kyoto (WKY) rats exhibit hypothyroidism and increased depressive and anxiety-like behaviors. Thus, the WKY could illuminate the mechanisms by which the reversal of developmental hypothyroidism in humans and animals results in adult behavioral improvement. We examined the outcome of maternal thyroxine (T4) treatment on thyroid hormone-regulated functions and adult behavior of the WKY offspring. Pregnant WKY dams completed gestation with and without T4 administration and their adult male offspring were tested. Measures included depressive and anxiety-like behaviors, and thyroid hormone (TH) concentrations in both plasma and specific brain regions. In addition, the expression of two proteins affecting thyroid hormone trafficking and metabolism, monocarboxylate transporter 8 (MCT-8) and iodothyronine deiodinase type III (Dio3), and of several behavior-altering molecules, glucocorticoid receptor (GR), prepro-thyrotropin releasing hormone (prepro-TRH) and corticotrophin releasing hormone (CRH), were determined in the hippocampus and amygdala of the offspring. Prenatal T4 treatment of WKYs did not affect adult depressive behavior but increased anxiety-like behavior and decreased plasma levels of THs. In the hippocampus of males treated with T4 in utero, Dio3 and MCT-8 protein levels were increased, while in the amygdala, there were increases of free T4, MCT-8, GR, prepro-TRH protein and CRH mRNA levels. These results show that T4 administration in utero programs adult peripheral and amygdalar thyroid hormone levels divergently, and that the resulting upregulation of anxiety-related genes in the amygdala could be responsible for the exacerbated anxiety-like behavior seen in WKYs after prenatal T4 treatment. PMID- 20005053 TI - Evolution, obesity, and why children so often choose the unhealthy eating option. AB - In recent decades obesity has become an increasing concern for governments and health organisations around the world. Changes in diet in modern conditions have led to our food environment being labelled "toxic" and "obesogenic", and are portrayed in marked contrast to the types of food environment in which humans evolved and are adapted to biologically. In this article I question whether the modern food environment is genuinely so different from those that humans evolved in, and whether domesticated animals, like humans, tend to become obese in conditions where food is plentiful. I continue by offering a model of eating behaviour based on the marginal value theorem. The model implies that many parents and public policy unwittingly encourage unhealthy diets by controlling or over-regulating children's diet: although it may feel counterintuitive to respond to the risk of obesity by allowing individuals to make choices rather than restricting their choices, a more relaxed and less regulated approach to food intake should allow appropriate modulation of food intake in conditions where food is plentiful. PMID- 20005054 TI - EEG-desynchronization as the major mechanism of anti-epileptic action of vagal nerve stimulation in patients with intractable seizures: clinical neurophysiological evidence. AB - Vagal nerve stimulation (VNS) has emerged as an effective and acceptable alternative therapy for patients with intractable seizures. Despite its reported efficacy in several studies, the precise mechanism of its anti-epileptic action remains elusive and requires to be established. Based on neurophysiological alterations induced by VNS, it has been proposed earlier that a major mechanism of its anti-epileptic action could be EEG-desynchronization, which is known to be associated with increased resistance to seizures. This brief paper attempts to support the proposed mechanism with clinical neurophysiological evidence that has emerged in recent times. PMID- 20005056 TI - Predictors of female urinary incontinence at midlife and beyond. AB - Urinary incontinence (UI) is an important middle age health issue and approximately 20% of women over 40 years of age have problems with continence. Urinary incontinence poses a significant negative impact on social functioning and quality of life to many individuals. It is estimated that around three million people are regularly incontinent in the UK with a prevalence of about 40 per 1000 adults. There are various factors which can cause incontinence such as pregnancy, childbirth, obesity, menopause, or just inherent connective-tissue weakness. All of these factors can cause pathophysiology changes in the muscular and fascial structures of the pelvic floor and lead to pelvic support defects and possibly pelvic floor dysfunction. We aim with this review article to highlight predictors or predisposing factors of incontinence; in order to help clinicians during their decisions and put in place a policy of a preventive strategy to decrease the incontinence rate in the general population. PMID- 20005055 TI - Detection and characterisation of O157:H7 and non-O157 Shiga toxin-producing Escherichia coli in wild boars. AB - The aim of this work was to determine the prevalence and characteristics of Escherichia coli O157:H7 and non-O157 Shiga toxin-producing E. coli (STEC) in free-ranging wild boars killed during the hunting season in southwest Spain. Faecal samples from 212 wild boars (Sus scrofa) were collected and examined for STEC. Characterisation of isolates was performed by PCR, serotyping, phage typing, and pulsed-field gel electrophoresis (PFGE). E. coli O157:H7 and non-O157 STEC were isolated from 7 (3.3%) and 11 (5.2%) animals, respectively, and the resulting 19 isolates were characterised. The PCR procedure indicated that 4 isolates carried the stx(1) gene, 12 carried the stx(2) gene, and 1 contained both of these genes. The ehxA, eae, and saa genes were detected in 13, 8, and 1 of the isolates, respectively. The eae-positive isolates comprised the types eae gamma 1 and eae-zeta. The isolates belonged to 11 O:H serotypes, including 4 new serotypes not previously reported within STEC strains, and the majority of them were from serotypes previously associated with human infection. E. coli O157:H7 isolates belonged to phage types associated with severe human illness: PT14, PT34, and PT54. Indistinguishable PFGE types were found in E. coli O157:H7 isolates recovered from a wild boar and from a human patient with diarrhoea living in the same geographic area. PMID- 20005057 TI - Comparison of adipocere formation in four soil types of the Porto (Portugal) district. AB - Four typical soils of the Porto (Portugal) area were characterized and used to study the decomposition of buried pieces of pork meat under controlled laboratory experiments (an 8 month experiment with a relatively high soil moisture and a 1 month experiment with relatively low soil moisture). The soils types were: organic, sandy, gravel and clay-gravel soils. Soils were characterized for their grain size distribution, pH, water content, organic matter percentage and mineral composition. Four free fatty acids (myristic, palmitic, oleic and stearic) were analysed (using a methodology based on an extraction step followed by a derivatization reaction and high performance liquid chromatography analysis) in soil samples as a sign of adipocere formation. The direct sensorial analysis of the buried sample residues and the free fatty acids profiles of the sampled soils showed that sandy and clay-gravel soils (in a low moisture environment) slowed the normal decomposition process promoting the formation of adipocere. Nevertheless, this apparent soil effect is indirect and a consequence of the different water retention and permeability of the soils. Thus, the water content of the soils is a crucial factor for adipocere formation. PMID- 20005058 TI - [Childhood pharyngitis in Sfax (Tunisia): epidemiology and utility of a rapid streptococcal test]. AB - OBJECTIVE: The authors studied the epidemiology of pharyngitis in children and evaluated the contribution of a rapid streptococcal test. PATIENTS AND METHODS: This prospective study was conducted from June 2007 to May 2008 in a primary health care institution (Policlinique de la Caisse Nationale de Securite Sociale) in Tunisia. Clinical findings were recorded; throat swabs were performed, and a rapid streptococcal test was made. RESULTS: Five hundred and four children were included in this study. The mean age was 5 years and 8 months. Culture was positive for group A streptococcus in 166 cases (32.9 %). Group A streptococcus was more frequently isolated in children aged 5 to 8 years. There was no difference in clinical features between groups of streptococcal and non streptococcal pharyngitis. The sensitivity and specificity of the rapid test were respectively 93.2 and 95.3 %. CONCLUSION: Pharyngitis was of bacterial origin in one third of cases; among these, group A streptococcus was the most frequent bacterium. The rapid test is a quick and reliable tool for the diagnosis of pharyngitis and hence an appropriate treatment. PMID- 20005060 TI - Comparison of transient arterial occlusion and muscle exercise provocation for assessment of perfusion reserve in skeletal muscle with real-time contrast enhanced ultrasound. AB - OBJECTIVE: Contrast-enhanced ultrasound (CEUS) is able to quantify muscle perfusion and changes in perfusion due to muscle exercise in real-time. However, reliable measurement of standardized muscle exercise is difficult to perform in clinical examinations. We compared perfusion reserve assessed by CEUS after transient arterial occlusion and exercise to find the most suitable measurement for clinical application. METHODS: Contrast pulse sequencing (7 MHz) during continuous IV infusion of SonoVue((r)) (4.8 mL/300 s) was used in 8 healthy volunteers to monitor muscle perfusion of the gastrocnemius muscle during transient (1 min) arterial occlusion produced by a thigh cuff of a venous occlusion plethysmograph. Isometric muscle exercise (50% of individual maximum strength for 20s) was subsequently performed during the same examination, and several CEUS parameters obtained from ultrasound-signal-intensity-time curves and its calculation errors were compared. RESULTS: The mean maximum local blood volume after occlusion was 13.9 [~mL] (range, 4.5-28.8 [~mL]), and similar values were measured after sub-maximum exercise 13.8 [~mL], (range, 4.6-22.2 [~mL]. The areas under the curve during reperfusion vs. recovery were also similar (515.2+/ 257.5 compared to 482.2+/-187.5 [~mLs]) with a strong correlation (r=0.65), as were the times to maximum (15.3s vs. 15.9s), with a significantly smaller variation for the occlusion method (+/-2.1s vs. +/-9.0s, p=0.03). The mean errors for all calculated CEUS parameters were lower for the occlusion method than for the exercise test. CONCLUSIONS: CEUS muscle perfusion measurements can be easily performed after transient arterial occlusion. It delivers data which are comparable to CEUS measurements after muscle exercise but with a higher robustness. This method can be easily applied in clinical examination of patients with e.g. PAOD or diabetic microvessel diseases to assess perfusion reserve. PMID- 20005059 TI - Secretin-stimulated MR cholangio-pancreatography in the evaluation of asymptomatic patients with non-specific pancreatic hyperenzymemia. AB - PURPOSE: To assess the diagnostic value of secretin-stimulated MRCP (SS-MRCP) compared with conventional MRCP in asymptomatic patients with mild elevations of pancreatic enzymes. MATERIALS AND METHODS: Eighty asymptomatic patients with pancreatic hyperenzymemia underwent MR imaging at 1.5T-device (Signa EXCITE, GE Healthcare). After the acquisition of axial T1w,T2w sequences, and conventional MRCP, SS-MRCP was performed using a single-slice coronal breath-hold, thick-slab, SSFSE T2w sequence, repeated every 30s up to 15 min following intravenous injection of secretin (Secrelux, Sanochemia). RESULTS: On the basis of the standards of reference, our final diagnoses were: negative findings (n=23), pancreas divisum (n=22), mild chronic pancreatitis (n=14), inflammatory ampullary stenosis (n=3), juxtapapillary duodenal diverticulum (n=1), small cystic lesions (<1cm) (n=22; 5/22 cases associated with pancreas divisum). The image quality of SS-MRCP was significantly higher than that of conventional MRCP (p<0.0001). Standards of reference did not differ significantly from of SS-MRCP findings (p=0.5), while was statistically different from those of conventional MRCP (p<0.0001). A significant difference was found between conventional MRCP and SS MRCP findings (p<0.0001). CONCLUSION: In asymptomatic patients with non-specific pancreatic hyperenzymemia SS-MRCP may represent the best non-invasive diagnostic technique, since it gives morphological and functional information. PMID- 20005061 TI - Contrast enhanced ultrasound of splenic lymphoma involvement. AB - OBJECTIVE: The aim of this study was to compare the value of contrast-enhanced ultrasonography (CEUS) with standard B-mode ultrasound (US) for diagnosis of splenic lymphoma involvement. METHODS: From 04/2005 to 10/2008 n=250 lymphoma patients were investigated by standard B-mode US. A homogeneous splenic echotexture was found in 199 patients (79%). To clarify the benefit of CEUS in this group a pilot series was performed with 16 of the 199 lymphoma patients. All patients with an abnormal splenic echotexture on standard B-Mode US (n=51) including focal hypoechoic splenic lesions (n=41) and an inhomogeneous splenic texture (n=10) were studied by CEUS. CEUS data were retrospectively evaluated. The diagnoses included indolent lymphoma (n=27), aggressive lymphoma (n=14), and Hodgkin's disease (n=10). Number and size of lesions were determined by B-mode US and CEUS. The visualisation of splenic lymphoma involvement by CEUS in comparison to B-mode US was classified as worse, equal, or better. RESULTS: All patients with a homogeneous spleen on B-mode US (n=16) had no visible focal lesions on CEUS. Study patients with focal lesions (n=41) had a hypoechoic (n=22) or isoechoic (n=19) enhancement during the arterial phase, and a hypoechoic enhancement during the parenchymal phase (n=41). The visualisation of focal splenic lymphoma was equal (n=32), better (n=6), or worse (n=3). In all study patients with an inhomogeneous spleen on B-mode US (n=10) no focal lesions were found by CEUS and the value of CEUS therefore was classified as worse. CONCLUSION: CEUS has no clear advantage for diagnosis of splenic lymphoma involvement. PMID- 20005062 TI - The effect of radiofrequency ablation on different organs: ex vivo and in vivo comparative studies. AB - OBJECTIVE: The purposes of this study are to evaluate the ex vivo and in vivo efficacy of radiofrequency ablation (RFA) on different porcine tissues by the ablation of three different sites simultaneously. MATERIALS AND METHODS: A multichannel RFA system, enables three separate tumors to be ablated simultaneously, was used. RFA procedures were applied to normal porcine liver, kidney, and muscle together ex vivo (n=12) and in vivo (n=17). Pre-impedances, defined as baseline systemic impedances of tissues before beginning RFA, and the areas of ablation zones were measured and compared. RESULTS: The areas of ablation zones among three organs had a significant difference in decreasing order as follows: liver, muscle, and kidney in the ex vivo study (p=0.001); muscle, liver, and kidney in the in vivo study (p<0.0001). The areas of ablation zones between ex vivo and in vivo had a significant difference in the liver and muscle (each p<0.05). There was no significant correlation between the areas of ablation zones and pre-impedances in both studies. CONCLUSIONS: Renal RFA produced the smallest ablation zone in both in vivo and ex vivo studies. Muscular RFA demonstrated the largest ablation zone in the in vivo study, and hepatic RFA showed the largest ablation zone in the ex vivo study. This variability in the tissues should be considered for performing an optimized RFA for each organ site. PMID- 20005063 TI - Characterization of human serum albumin forms with pH. Fluorescence lifetime studies. AB - Fluorescence lifetimes of human serum albumin (HSA) tryptophan 214 residue were measured in solution at different pH (from 2 to 12). The results indicate that tryptophan emission occurs with three lifetimes at all pH. However, lifetimes and pre-exponential values are dependent on the pH and thus on the protein form. Three different protein populations have been differentiated: one population for pH 2 and 3 (extended form), the second one from pH 4 to 9 containing HSA migrating (F), normal (N) and basic (B) forms. Another type of population is obvious for pH higher than 9, characterizing the aged (A) form of HSA. PMID- 20005064 TI - A comparative characterization of dipentameric (IgM)(2) and pentameric IgM species present in preparations of a monoclonal IgM for therapeutic use. AB - IgM aggregates in biotechnologically produced preparations have been reported, however, in vitro characteristics and in vivo activity of IgM aggregates have not been well studied. We separated two species of the human monoclonal IgM antibody KBPA-101 by size-exclusion chromatography. Molecular weight determination indicated the presence of dipentameric and pentameric forms. We present here the results of a comparative characterization of these IgM species, including in vitro and in vivo effector function against Pseudomonas aeruginosa. Dipentameric (IgM)(2) species were observed to dissociate into pentameric IgM at 37 degrees C, suggesting a dynamic equilibrium, in which the pentameric species is the predominant form. In vitro antigen binding (P. aeruginosa LPS) and IgM-mediated complement-dependent phagocytosis of labeled bacterial cells did not differ significantly between the dipentameric (IgM)(2) and pentameric IgM species. Furthermore, the in vivo efficacy of dipentameric and pentameric IgM in protecting mice from a lethal dose of P. aeruginosa through passive immunization was nearly equivalent. In conclusion, low concentrations of dipentameric (IgM)(2) may contain an additional but equally active component of the principal biological form. The data presented in this work support the conclusion that the pentameric form of IgM directed against the O-polysaccharide moiety of P. aeruginosa serotype IATS-O11 and dipentameric (IgM)(2) are functionally equivalent. PMID- 20005065 TI - Rifampicin determination in plasma by stir bar-sorptive extraction and liquid chromatography. AB - A sensitive and reproducible stir bar-sorptive extraction and high performance liquid chromatography-UV detection (SBSE/HPLC-UV) method for therapeutic drug monitoring of rifampicin in plasma samples is described and compared with a liquid:liquid extraction (LLE/HPLC-UV) method. This miniaturized method can result in faster analysis, higher sample throughput, lower solvent consumption and less workload per sample while maintaining or even improving sensitivity. Important factors in the optimization of SBSE efficiency such as pH, temperature, extraction time and desorption conditions (solvents, mode magnetic stir, mode ultrasonic stir, time and number of steps) were optimized recoveries ranging from 75 to 80%. Separation was obtained using a reverse phase C(8) column with UV detection (254nm). The mobile phase consisted of methanol:0.25N sodium acetate buffer, pH 5.0 (58:42, v/v). The SBSE/HPLC-UV method was linear over a working range of 0.125-50.0microgmL(-1). The intra-assay and inter-assay precision and accuracy were studied at three concentrations (1.25, 6.25 and 25.0microgmL(-1)). The intra-assay coefficients of variation (CVs) for all compounds were less than 10% and all inter-CVs were less than 10%. Limits of quantification were 0.125microgmL(-1). Stability studies showed rifampicin was stable in plasma for 12h after thawing; the samples were also stable for 24h after preparation. Based on the figures of merit results, the SBSE/HPLC-UV proved to be adequate to the rifampicin analyses from therapeutic to toxic levels. This method was successfully applied to the analysis of real samples and was as effective as the LLE/HPLC-UV method. PMID- 20005066 TI - Emotional and informational patient cues: the impact of nurses' responses on recall. AB - OBJECTIVE: To investigate older cancer patients' informational and emotional cues, how nurses respond to these cues and the effect of cues and responses on patients' information recall. METHODS: 105 cancer patients (aged >or=65 years) completed a recall questionnaire after an educational session preceding chemotherapy treatment. Recall was checked against the actual communication in videorecordings of the consultations. Patients' emotional and informational cues and subsequent responses by the nurse were rated using an adaptation of the Medical Interview Aural Rating Scale (MIARS). RESULTS: Patients gave more informational than emotional cues. The most frequent response to emotional cues was distancing followed by acknowledgement. Nurses gave appropriate information in response to the majority of informational cues. Patients' expression of emotional or informational cues did not influence recall; neither did nurses' responses to informational cues. Responses to emotional cues did affect recall. The more nurses responded by giving 'minimal' encouragements (e.g. 'Hmmm'), the more patients recalled, while distancing responses (e.g. switching focus) were associated with lower recall scores. CONCLUSION: Responding to patients' emotions is likely to impact information recall. PRACTICE IMPLICATIONS: These results highlight the importance of addressing patients' expressions of emotions in the context of patient education, as it enhances information recall. PMID- 20005067 TI - What constitutes evidence-based patient information? Overview of discussed criteria. AB - OBJECTIVE: To survey quality criteria for evidence-based patient information (EBPI) and to compile the evidence for the identified criteria. METHODS: Databases PubMed, Cochrane Library, PsycINFO, PSYNDEX and Education Research Information Center (ERIC) were searched to update the pool of criteria for EBPI. A subsequent search aimed to identify evidence for each criterion. Only studies on health issues with cognitive outcome measures were included. Evidence for each criterion is presented using descriptive methods. RESULTS: 3 systematic reviews, 24 randomized-controlled studies and 1 non-systematic review were included. Presentation of numerical data, verbal presentation of risks and diagrams, graphics and charts are based on good evidence. Content of information and meta information, loss- and gain-framing and patient-oriented outcome measures are based on ethical guidelines. There is a lack of studies on quality of evidence, pictures and drawings, patient narratives, cultural aspects, layout, language and development process. CONCLUSION: The results of this review allow specification of EBPI and may help to advance the discourse among related disciplines. Research gaps are highlighted. PRACTICE IMPLICATIONS: Findings outline the type and extent of content of EBPI, guide the presentation of information and describe the development process. PMID- 20005068 TI - Gene expression profile towards the prediction of patient survival of gastric cancer. AB - BACKGROUND: Several gene expression signatures have been reported to predict patient survival of gastric cancer after surgical resection. However, the prognostic gene lists have overlapped poorly until now. This study conducted an analysis to characterize gene expression profile and developed a survival prediction model. METHODS: The gene expression profile was evaluated in fresh frozen tumor tissue obtained from 48 patients with primary gastric cancer. We measured 84 representative genes involved in transformation and tumorigenesis using quantitative reverse transcription polymerase chain reaction (qRT-PCR) and related the results to overall survival. RESULTS: In a univariate analysis, 84 genes were ranked on their ability to predict survival, of which nine genes were the strongest predictor (P<0.05). They were PLAU, MAP2K1, THBS1, TWIST1, ITGB5, NME4, ANGPT2, platelet-derived growth factor B (PDGFB), ITGB1. Then, we did a multivariate analysis to further select four genes (ITGB1, PDGFB, THBS1, TWIST1) from the above nine genes for the construction of biomathematics model, which was independent of age, gender, TNM stage and other variables. This model could correctly clarify gastric patients into the high-risk group, median-risk group and low-risk group, as well as predict their survival. CONCLUSIONS: Measurement of the expression of four genes is probable to predict surgery-related survival. This model may be test further for its potential to improve the selection of the resected gastric cancer patients in adjuvant chemotherapy. PMID- 20005069 TI - Combined tamoxifen and gefitinib in non-small cell lung cancer shows antiproliferative effects. AB - Gefitinib, an epidermal growth factor receptor (EGFR) tyrosine kinase inhibitor, is approved for clinical use in the treatment of non-small cell lung cancer (NSCLC). According to statistics, NSCLC patients who are female, have adenocarcinoma, or never smoked have a higher response rate to gefitinib treatment. This phenomenon could be due to the interaction between the estrogen receptor (ER) and EGFR. To test whether inhibition of the EGFR signaling pathway affects the antitumour effect of gefitinib, NSCLC cell lines were treated with gefitinib and tamoxifen, an ER antagonist. Cotreatment with gefitinib plus tamoxifen decreased the proliferation and increased the apoptosis of A549 and H1650 adencarcinoma cell lines, when compared with either drug alone. However, there was no effect on H520 cells (squamous cell carcinoma). Rapid activation of the EGFR pathway by both EGF and beta-E2 was observed in A549 cells. Additionally, EGFR and ERbeta expression was down-regulated in response to estrogen and EGF, respectively, but up-regulated in response to tamoxifen and genfitib, respectively. These results suggest that there is a functional cross signaling between the EGFR and the ER pathways in NSCLC, possibly providing a rationale to combine gefitinib with anti-estrogen therapy for lung cancer treatment. PMID- 20005070 TI - [Is there a doctor on board?]. AB - Medical problems during flight have become an important issue as the number of passengers and of miles flown continue to rise. Cabin pressurization causes hypoxia, hypobaria and decreased humidity, which are responsible for most medical incidents occurring during flight. Worldwide daily medical incidents are estimated at 350, i.e., one per 14,000 to 39,600 passengers. Medical advice is obtained in 69% of cases, from physicians (40%), nurses (25%), or paramedics (4%) on board the plane. The leading causes of medical incidents are gastrointestinal (25%), cardiac (10%), and neurological (10%) diseases. The incidence of cardiac arrest in flight is 1000 cases a year, worldwide. Since 1992, airlines have progressively equipped their planes with automated external defibrillators, and crew members are trained to use them. Passenger flights carry medical equipment and drugs, determined according to number of passengers and the flight distance. The conditions of intervention are codified: "good Samaritan" laws protect professionals from liability when they choose to aid others who are injured or ill. Current recommendations call for physicians to identify themselves, request an interpreter when necessary, obtain the patient's consent, conduct out examination, inform the patient, family members and crew members of the situation, contact ground medical staff, use well-known procedures, consider flight diversion, and write up a case report. PMID- 20005071 TI - [New Influenza A H1N1v: usefulness of immunization]. PMID- 20005072 TI - A fast algorithm for AR parameter estimation using a novel noise-constrained least-squares method. AB - In this paper, a novel noise-constrained least-squares (NCLS) method for online autoregressive (AR) parameter estimation is developed under blind Gaussian noise environments, and a discrete-time learning algorithm with a fixed step length is proposed. It is shown that the proposed learning algorithm converges globally to an AR optimal estimate. Compared with conventional second-order and high-order statistical algorithms, the proposed learning algorithm can obtain a robust estimate which has a smaller mean-square error than the conventional least squares estimate. Compared with the learning algorithm based on the generalized least absolute deviation method, instead of minimizing a non-smooth linear L(1) function, the proposed learning algorithm minimizes a quadratic convex function and thus is suitable for online parameter estimation. Simulation results confirm that the proposed learning algorithm can obtain more accurate estimates with a fast convergence speed. PMID- 20005073 TI - The complement system in systemic autoimmune disease. AB - Complement is part of the innate immune system. Its major function is recognition and elimination of pathogens via direct killing and/or stimulation of phagocytosis. Activation of the complement system is, however, also involved in the pathogenesis of the systemic autoimmune diseases. Activation via the classical pathway has long been recognized in immune complex-mediated diseases such as cryoglobulinemic vasculitis and systemic lupus erythematosus (SLE). In SLE, the role of complement is somewhat paradoxical. It is involved in autoantibody-initiated tissue damage on the one hand, but, on the other hand, it appears to have protective features as hereditary deficiencies of classical pathway components are associated with an increased risk for SLE. There is increasing evidence that the alternative pathway of complement, even more than the classical pathway, is involved in many systemic autoimmune diseases. This is true for IgA-dominant Henoch Schonlein Purpura, in which additional activation of the lectin pathway contributes to more severe disease. In anti-neutrophil cytoplasmic antibody (ANCA)-associated vasculitis the complement system was considered not to be involved since immunoglobulin deposition is generally absent in the lesions. However, recent studies, both in human and animal models, demonstrated complement activation via the alternative pathway as a major pathogenic mechanism. Insight into the role of the various pathways of complement in the systemic autoimmune diseases including the vasculitides opens up new ways of treatment by blocking effector pathways of complement. This has been demonstrated for monoclonal antibodies to C5 or C5a in experimental anti phospholipid antibody syndrome and ANCA-associated vasculitis. PMID- 20005074 TI - Recurring gnathodiaphyseal dysplasia in two Russian brothers. AB - Two Russian brothers presented with recurring benign facial bone tumors and progressive limb bowing. The association of fibro-osseous jawbone lesions and long-bone bowing with cortical thickening suggested the diagnosis of gnathodiaphyseal dysplasia, in the absence of arguments in favor of fibrous dysplasia. Gnathodiaphyseal dysplasia is a rare autonomic dominant syndrome due to a mutation of the TMEM16E gene. The extreme and recurring phenotype of these two patients illustrates the variable expressivity of this disease. Differential diagnosis with other benign facial bone tumors is discussed. PMID- 20005075 TI - Paradental cyst mimicking a periodontal pocket: case report of a conservative treatment approach. AB - A 7-year-old boy presented with a periodontal problem related to an erupting lower molar. The tooth showed a 15 mm deep periodontal pocket on the buccal aspect. A microbiological DNA test excluded a periodontal origin. The treatment consisted of local antimicrobial therapy and cleaning and filling of the pocket with Atridox. 2 years after therapy the pocket completely disappeared. Finding periodontal pockets on freshly erupted teeth with acute symptoms should suggest the diagnosis of a cyst. This could prevent surgical endodontal or periodontal therapy. This problem can be managed effectively with minimal therapy and local antibiotics. PMID- 20005076 TI - Reactive lesions of peri-implant mucosa associated with titanium dental implants: a report of 2 cases. AB - The aim of this study was to report 2 novel clinical cases of reactive lesions of the peri-implant mucosa associated with titanium dental implants where metal-like particles were observed histologically. In both cases, the lesions were diagnosed as epulis, based on clinical evidence. Extirpation biopsies were carried out. Case 1 was diagnosed as pyogenic granuloma and case 2 as peripheral giant cell granuloma. The presence of metal-like particles in the tissues suggests that the etiology of the lesions might be related to the corrosion process of the metal structure. This is the first case of pyogenic granuloma to be reported in association with dental implants. All clinical cases of soft tissue lesions associated with implants should be reported to contribute to the understanding of the etiology and pathogeny of these lesions. PMID- 20005077 TI - Febrile seizures and temporal lobe epileptogenesis. AB - Febrile seizures (FS) are a common neurological disorder that affects children. Simple FS are thought to be benign but experimental and clinical evidence support that the risk of developing epilepsy after FS increases if the FS are prolonged and the brain is abnormal. In addition, prolonged FS (PFS) have many deleterious long-term effects characterized mainly in the hippocampus but may involve the whole brain and that prompt abortive treatment of PFS may prevent some of the adverse effects. This review focuses on some of the key factors involved in the generation of FS, factors leading to PFS and potential mechanisms and functional correlates leading to temporal lobe epilepsy (TLE). PMID- 20005078 TI - Sleep and epilepsy in children: clinical aspects and polysomnography. AB - Studies assessing the relationship between sleep and epilepsy in pediatric patients are scarce, however, during childhood, the incidence either of epilepsy and sleep disorders is elevated and this population is thought to have the greatest occurrence of the association between these two co-morbidities. In this article we review results from clinical research developed in our sleep laboratory as effects of refractory partial epilepsy in sleep organization, sleep habits in children with epilepsy, the relationship of apparent life threatening events and epilepsy, heart rate control during sleep and seizures and risk of sudden death, and discuss our findings in the context of the literature available. PMID- 20005079 TI - Response to drug treatment in newly diagnosed epilepsy: a pilot study of (1)H NMR and MS-based metabonomic analysis. AB - Understanding the biological basis of drug resistance and developing techniques which facilitate prediction of outcome have the potential to revolutionize the pharmacotherapy of epilepsy. We have performed a pilot study of metabonomic analysis using nuclear magnetic resonance (NMR) spectroscopy and mass spectrometry (MS) in an effort to identify metabolic biomarkers of response to antiepileptic drug treatment. Pre-treatment serum samples were obtained from 125 patients with newly diagnosed epilepsy who were taking part in a randomised monotherapy trial. Outcome (responder, non-responder) was assessed at 6 weeks, 6 months, and 12 months after starting treatment. Serum samples were subject to investigation by both NMR and MS and the resulting data interrogated by principal components analysis (PCA). There was no clear distinction in the metabolic profile, acquired by either NMR or MS, of responders and non-responders to AED treatment at any of the three clinical end-points investigated, suggesting that pre-treatment serum samples do not contain any prominent biomarkers of responsiveness to initial treatment in new-onset epilepsy. Metabonomic analysis is undoubtedly applicable to the search for biological predictors of response to drug treatment in epilepsy, but future studies should employ larger patient cohorts, more discriminatory analyses, and a less equivocal clinical phenotype. PMID- 20005080 TI - Prevalence of metabolic syndrome in Japanese psoriasis patients. PMID- 20005081 TI - Evaluation of the effect of selective serotonin-reuptake inhibitors on lymphocyte subsets in patients with a major depressive disorder. AB - To date, only the effect of a short-term antidepressant treatment (<12 weeks) on neuroendocrinoimmune alterations in patients with a major depressive disorder has been evaluated. Our objective was to determine the effect of a 52-week long treatment with selective serotonin-reuptake inhibitors on lymphocyte subsets. The participants were thirty-one patients and twenty-two healthy volunteers. The final number of patients (10) resulted from selection and course, as detailed in the enrollment scheme. Methods used to psychiatrically analyze the participants included the Mini-International Neuropsychiatric Interview, Hamilton Depression Scale and Beck Depression Inventory. The peripheral lymphocyte subsets were measured in peripheral blood using flow cytometry. Before treatment, increased counts of natural killer (NK) cells in patients were statistically significant when compared with those of healthy volunteers (312+/-29 versus 158+/-30; cells/mL), but no differences in the populations of T and B cells were found. The patients showed remission of depressive episodes after 20 weeks of treatment along with an increase in NK cell and B cell populations, which remained increased until the end of the study. At the 52nd week of treatment, patients showed an increase in the counts of NK cells (396+/-101 cells/mL) and B cells (268+/-64 cells/mL) compared to healthy volunteers (NK, 159+/-30 cells/mL; B cells, 179+/-37 cells/mL). We conclude that long-term treatment with selective serotonin-reuptake inhibitors not only causes remission of depressive symptoms, but also affects lymphocyte subset populations. The physiopathological consequence of these changes remains to be determined. PMID- 20005082 TI - Fabrication of cell pattern on poly(dimethylsiloxane) by vacuum ultraviolet lithography. AB - Cell patterning on substrates has played a significant role in the study of basic biology, cell-based biosensor and tissue engineering. In this report, a cell pattern was prepared on poly(dimethylsiloxane) (PDMS) substrate by vacuum ultraviolet (VUV) lithography. After immobilizing allyl-polyethylene glycol (APEG) onto PDMS, a chemical heterogeneous patterned surface was fabricated by VUV (Xe(2) excimer: 172nm) lithography with copper mesh as a photomask. The UV exposed domains can promote L929 cell adhesion and growth. However, non-exposed regions resist cell attachment because of the repelling property of PEG. Therefore, cell pattern could be achieved without pre-adsorption of cell adhesive species before cell culture. PMID- 20005083 TI - Effect of surface area enhancement on the adsorption of Bovine Serum Albumin onto titanium dioxide. AB - The equilibrium and the kinetics of BSA adsorption onto specific surface area changed TiO(2) by heat treatment were studied. The TiO(2) was treated at 100 and 200 degrees C for 24h. The specific surface areas of TiO(2) characterized by BET method were measured as 48.9m(2)/g and 53.0m(2)/g for 100 and 200 degrees C, respectively. The adsorption rate and the equilibrium experiments were carried out at pH 4 for 40 degrees C. The rate kinetics for the adsorption of BSA was best fitted with the pseudo-first-order kinetic model. The equilibrium process was described by the Langmuir and Freundlich isotherm models. The adsorption capacities (Q(0)) calculated from the Langmuir isotherm model were 40.6 and 44.4mg/g for heat-treated TiO(2) at 100 and 200 degrees C, respectively. The adsorption of BSA increased with increasing surface area of TiO(2). The zeta potential values of the 100- and 200 degrees C-treated TiO(2) were found as 2.57mV and 0.39mV, respectively, showing that the interaction between TiO(2) with BSA increased with increasing temperature of heat treatment for TiO(2). PMID- 20005084 TI - Editorial: Lymphatic vessel functions in Health and Disease. PMID- 20005085 TI - Possible effect of gene polymorphisms on the release of TNFalpha and IL1 cytokines in coal workers' pneumoconiosis. AB - It has been shown that coal dust exposure stimulates inflammatory response leading to increased release of cytokines from monocytes such as TNF-alpha and IL1. These released cytokines play the key role in the pathogenesis of pneumoconiosis including coal workers' pneumoconiosis. In this study, we investigated TNFA, IL1A, IL1B and IL1RA genes variations on basal, lipopolysaccharide and coal dust-induced cytokine release from blood monocytes of homozygous allele and minor variant allele carriers in Turkish coal workers and CWP patients. According to the genotyping results, TNFA -238 gene polymorphism was found as a risk factor in CWP development (OR=3.79) and to in vitro results; release of both TNF-alpha and IL1 cytokines from the monocytes in CWP patients was significantly increased compared to the healthy workers. Also, LPS and coal dust stimulated release of TNF-alpha, which was significantly higher in allele 2 carriers compared to subjects carrying allele 1 in both the groups. These data suggest that the coal dust-induced release of TNF-alpha from monocytes may be a useful biomarker of CWP. PMID- 20005086 TI - Co-existence of COPD and left ventricular dysfunction in vascular surgery patients. AB - BACKGROUND: The co-existence between chronic obstructive pulmonary disease (COPD) and heart failure has been previously described. However, the co-existence between COPD and subclinical left ventricular (LV) dysfunction, without the presence of heart failure symptoms, is less well understood. This study determined the relationship and clinical relevance of COPD and subclinical LV dysfunction in vascular surgery patients. METHODS: 1005 consecutive vascular surgery patients were included in which COPD was determined using spirometry and LV function using echocardiography. Mild COPD was defined as FEV(1)>or=80% of predicted+FEV(1)/FVC-ratio<0.70. Moderate/severe COPD was defined as FEV(1)<80% of predicted+FEV(1)/FVC-ratio<0.70. Systolic LV dysfunction was defined as LV ejection fraction <50% and diastolic LV dysfunction was diagnosed based on E/A ratio, pulmonary vein flow and deceleration time. Multivariate regression analyses were used to evaluate the impact of COPD and LV dysfunction on all-cause mortality. The mean follow-up time was 2.2+/-1.8 years. RESULTS: Both, mild and moderate/severe COPD were associated with increased risk for subclinical LV dysfunction with odds ratio of 1.6 (95%-CI=1.1-2.3) and 1.7 (95%-CI=1.2-2.4), respectively. Mild- or moderate/severe COPD in combination with LV dysfunction was associated with increased risk for all-cause mortality (mild: hazard ratio 1.7; 95%-CI=1.1-3.6, moderate/severe: hazard ratio 2.5; 95%-CI=1.5-4.7). CONCLUSIONS: COPD was associated with increased risk for subclinical LV dysfunction. COPD+subclinical LV dysfunction was associated with increased risk for all-cause mortality compared to patients with COPD+normal LV function. Echocardiography may be useful to detect subclinical cardiovascular disease and risk-stratify COPD patients undergoing vascular surgery. PMID- 20005087 TI - Chronic cough in upper airway diseases. AB - BACKGROUND: The epidemiological, pathophysiological and clinical links between upper and lower airways are nowadays clearly demonstrated. Most of asthmatics are suffering from rhinitis while up to 40% of rhinitic patients have asthma. Asthmatics and COPD patients are also prone to develop concomitant chronic rhinosinusitis (CRS). This study aimed to determine the predictive value of cough for concomitant asthma in patients suffering from upper airway diseases. METHODS: This cross-sectional study described a group of 143 consecutive patients suffering simultaneously from common upper and lower airway disorders. Both ENT specialists and respiratory physicians consecutively examined the patients in Ghent University Hospital from October 2004 till October 2006. This study was based on the demographic characteristics, upper and lower airway conditions. RESULTS: Forty-seven percent of the patients included in the study were males and the mean age of studied population was 43.6 years. The major complaint was chronic cough. When present, patients with chronic cough have an increased risk of suffering from a concomitant asthma in both allergic rhinitis (OR=5.8) and CRS with nasal polyps (OR=10.4), but not in CRS without polyps. CONCLUSIONS: Chronic cough was found to be a key symptom of associated asthma in allergic rhinitis and CRS with nasal polyps. Interestingly, chronic cough in CRS without nasal polyps did not show the same predictive value: this suggests different pathophysiological mechanisms. PMID- 20005088 TI - Comparative metabolomics approach coupled with cell- and gene-based assays for species classification and anti-inflammatory bioactivity validation of Echinacea plants. AB - Echinacea preparations were the top-selling herbal supplements or medicines in the past decade; however, there is still frequent misidentification or substitution of the Echinacea plant species in the commercial Echinacea products with not well chemically defined compositions in a specific preparation. In this report, a comparative metabolomics study, integrating supercritical fluid extraction, gas chromatography/mass spectrometry and data mining, demonstrates that the three most used medicinal Echinacea species, Echinacea purpurea, E. pallida, and E. angustifolia, can be easily classified by the distribution and relative content of metabolites. A mitogen-induced murine skin inflammation study suggested that alkamides were the active anti-inflammatory components present in Echinacea plants. Mixed alkamides and the major component, dodeca-2E,4E,8Z,10Z(E) tetraenoic acid isobutylamides, were then isolated from E. purpurea root extracts for further bioactivity elucidation. In macrophages, the alkamides significantly inhibited cyclooxygenase 2 (COX-2) activity and the lipopolysaccharide-induced expression of COX-2, inducible nitric oxide synthase and specific cytokines or chemokines [i.e., TNF-alpha, interleukin (IL)-1alpha, IL-6, MCP-1, MIP-1beta] but elevated heme oxygenase-1 protein expression. Cichoric acid, however, exhibited little or no effect. The results of high-performance liquid chromatography/electron spray ionization/mass spectrometry metabolite profiling of alkamides and phenolic compounds in E. purpurea roots showed that specific phytocompound (i.e., alkamides, cichoric acid and rutin) contents were subject to change under certain post-harvest or abiotic treatment. This study provides new insight in using the emerging metabolomics approach coupled with bioactivity assays for medicinal/nutritional plant species classification, quality control and the identification of novel botanical agents for inflammatory disorders. PMID- 20005090 TI - Systematic review and meta-analysis of the used surgical techniques to reduce leg lymphedema following radical inguinal nodes dissection. AB - BACKGROUND: Inguinal nodes dissection is associated with high rates of morbidity, lymphedema in particular is a chronic disabling condition which is a common complication following this operation. Prevention or minimization of this condition is an important aim when considering this procedure. Many technical modifications are suggested for this purpose. This systematic review aims at assessing the efficacy of the available strategies to reduce the risk and severity of leg lymphedema. METHODS: For this review, MEDLINE and EMBASE were searched to identify studies that reported surgical strategies designed to reduce complications of groin dissection and in particular leg lymphedema. Studies that reported outcome of long saphenous vein sparing, fascia preserving dissection, microvascular surgery, sartorius transposition and omental pedicle flap were located. Data were collected using predefined inclusion and exclusion criteria. A combined odds ratio was calculated combining studies suitable for meta-analysis using the random effect model. RESULTS: The search result defined few studies that reported results of saphenous vein sparing technique; some of those studies were found suitable for meta-analysis based on the Newcastle-Ottawa scale for non randomized studies. The meta-analysis showed significant reduction of lymphedema (odds ratio 0.24, 95% CI 0.11-0.53) and other complications of inguinal node dissection. There were no randomized studies to address this problem; there are also isolated studies that reported benefits of other techniques but none of them was suitable for meta-analysis. CONCLUSION: Meta-analysis of the reported studies on sparing the long saphenous vein in inguinal nodes dissection suggests a reduced rate of lymphedema and other postoperative complications. Other methods that may be beneficial are fascia preserving dissection, pedicled omental flap and microsurgery; however sartorius transposition has not been shown to reduce the rate of complications. Randomized controlled trials are needed to prove the benefits of various technical modifications. PMID- 20005089 TI - Tumour CD133 mRNA expression and clinical outcome in surgically resected colorectal cancer patients. AB - BACKGROUND: Human prominin-1 (CD133) is a novel pentaspan membrane protein which was originally classified as a marker of primitive haematopoietic and neural stem cells. Cancer stem cells have been isolated and expanded from leukaemia and several solid tumours, and have been associated with metastasis, chemoresistance and relapse. CD133 is recognised as a stem cell marker and is capable of identifying a tumour-initiating subpopulation in brain, colon, melanoma and other solid tumours. METHODS: We assessed CD133 mRNA expression levels by RT-QPCR in tumour and matched normal tissue from 64 stages I-III colorectal cancer (CRC) patients and correlated tumour CD133 levels with clinicopathological characteristics and clinical outcome. RESULTS: In four patients, CD133 mRNA was not expressed in tumour or in normal tissue. In the remaining 60 patients, expression levels were higher in tumour than in normal tissue (p=0.001). Higher levels of CD133 expression were associated with shorter relapse-free interval (RFI) (p=0.004) and overall survival (OS) (p<0.0001). In the multivariate analyses, CD133 levels emerged as a prognostic marker for RFI and OS. CONCLUSIONS: We have observed longer RFI and OS in patients with lower levels of CD133, regardless of adjuvant treatment and other clinical characteristics. If these findings are confirmed in larger prospective studies, CD133 assessment may prove useful for new diagnostic and therapeutic procedures for CRC patients. PMID- 20005091 TI - Characterization of an anaerobic baffled reactor treating dilute aircraft de icing fluid and long term effects of operation on granular biomass. AB - Successful treatment of dilute ethylene glycol based-aircraft de-icing fluid (ADF) was achieved using a four compartment, anaerobic baffled reactor (ABR). Three ADF concentrations (0.04, 0.07, and 0.13%v/v) were continuously fed at different hydrological retention times (HRTs; 24, 12, 6 and 3h) with concomitant organic loading rates (OLRs) varying between 0.3 and 6 kg chemical oxygen demand (COD)/m(3)/d. ABR achieved over 75% soluble COD removal and an average methane production potential of 0.30+/-0.05LCH(4)/gCOD(removed) at 33 degrees C for the experimental conditions evaluated. The different experimental conditions tested and a four-month summer shut-down simulation had no significant effect on reactor performance or on the settling characteristics of the granular biomass, which remained almost constant during the study. Biomass specific acetoclastic activity however, changed through the study; increasing two fold for the last three compartments and decreasing almost the same magnitude for the first compartment compared to inoculum, suggesting that a new distribution of microbial consortia was established in each compartment of the reactor by the end of the study. PMID- 20005092 TI - Extracellular terpenoid hydrocarbon extraction and quantitation from the green microalgae Botryococcus braunii var. Showa. AB - Mechanical fractionation and aqueous or aqueous/organic two-phase partition approaches were applied for extraction and separation of extracellular terpenoid hydrocarbons from Botryococcus braunii var. Showa. A direct spectrophotometric method was devised for the quantitation of botryococcene and associated carotenoid hydrocarbons extracted by this method. Separation of extracellular botryococcene hydrocarbons from the Botryococcus was achieved upon vortexing of the micro-colonies with glass beads, either in water followed by buoyant density equilibrium to separate hydrocarbons from biomass, or in the presence of heptane as a solvent, followed by aqueous/organic two-phase separation of the heptane solubilized hydrocarbons (upper phase) from the biomass (lower aqueous phase). Spectral analysis of the upper heptane phase revealed the presence of two distinct compounds, one absorbing in the UV-C, attributed to botryococcene(s), the other in the blue region of the spectrum, attributed to a carotenoid. Specific extinction coefficients were developed for the absorbance of triterpenes at 190nm (epsilon = 90 +/- 5 mM(-1) cm(-1)) and carotenoids at 450 nm (epsilon=165+/-5mM(-1) cm(-1)) in heptane. This enabled application of a direct spectrophotometric method for the quantitation of water- or heptane-extractable botryococcenes and carotenoids. B. braunii var. Showa constitutively accumulates approximately 30% of the dry biomass as extractable (extracellular) botryococcenes, and approximately 0.2% of the dry biomass in the form of a carotenoid. It was further demonstrated that heat-treatment of the Botryococcus biomass substantially accelerates the rate and yield of the extraction process. Advances in this work serve as foundation for a cyclic Botryococcus growth, non toxic extraction of extracellular hydrocarbons, and return of the hydrocarbon depleted biomass to growth conditions for further product generation. PMID- 20005093 TI - Characterization of vinasses from five certified brands of origin (CBO) and use as economic nutrient for the xylitol production by Debaryomyces hansenii. AB - Vinasses coming from the five CBOs of Galicia, north-western Spain, were characterized, and successfully employed as economic nutritional supplements for xylitol production by Debaryomyces hansenii. All fermentations can be modelled showing kinetic patterns fairly described by the mathematical models. No negative effect of the phenolic compounds in the liquid phase on the initial volumetric rate of product formation (r(P)(0)) was observed. Multiple linear regression analysis was used to describe the effect of metals and initial xylose acting on P(max) and Y(P/S). Zn was the most influential variable. Besides, partial least squares regression models show a clear separation, based on the first two principal components, between the whole vinasses and the liquid fractions, which provided the higher P(max), with the exception of CBO 4, where P(max)=40.4 g/L, was achieved using the solid and liquid fraction. PMID- 20005094 TI - Ethanolysis of rapeseed oil - distribution of ethyl esters, glycerides and glycerol between ester and glycerol phases. AB - The distribution of ethyl esters, triglycerides, diglycerides, monoglycerides, and glycerol between the ester and glycerol phase was investigated after the ethanolysis of rapeseed oil at various reaction conditions. The determination of these substances in the ester and glycerol phases was carried out by the GC method. The amount of ethyl esters in the glycerol phase was unexpectedly high and therefore the possibility of the reduction of this amount was investigated. The distribution coefficients and the weight distributions of each investigated substance were calculated and compared mutually. The distribution coefficients between the ester and glycerol phase increase in this sequence: glycerol, monoglycerides, diglycerides, ethyl esters, and triglycerides. Soaps and monoglycerides in the reaction mixture cause a worse separation of ethyl esters from the reaction mixture. The existence of a non-separable reaction mixture was observed also, and its composition was determined. PMID- 20005095 TI - Column studies on the evaluation of novel spacer granules for the removal of arsenite and arsenate from contaminated water. AB - Decontamination of arsenic ions from aqueous media has been investigated using iron chitosan spacer granules (ICS) as an adsorbent. Drying of beads saturated with a spacer sucrose was considered as simple treatment, to prevent the restriction of polymer network and enhance sorption capacity. The novel sorbent was studied in up flow column experiments conducted at different flow rates, pH and bed depth to quantify the treatment performance. It was found that silicate was more inhibitory than phosphate, and the silicate in groundwater controlled the arsenic removal efficiency. The column regeneration studies were carried out for two sorption-desorption cycles using 0.1N NaOH as the eluant. TCLP leaching tests were conducted on the arsenic loaded adsorbent which revealed the containment of arsenic-laden sludge can be managed without adverse environmental impact. The developed procedure was successfully applied for the removal of both As(III) and As(V) from arsenic contaminated drinking water samples. PMID- 20005096 TI - The identification of structurally novel, selective, orally bioavailable positive modulators of mGluR2. AB - The optimisation of an HTS hit series (1) leading to the identification of structurally novel, selective, orally bioavailable mGluR2 positive modulators GSK1331258 and GSK1331268 is described. Structure-activity relationships, attenuation of dopaminergic activity, and potentiation of mGluR2 responses in rat hippocampal MPP-DG synapses are also reported. PMID- 20005097 TI - Discovery of (pyridin-4-yl)-2H-tetrazole as a novel scaffold to identify highly selective matrix metalloproteinase-13 inhibitors for the treatment of osteoarthritis. AB - Potent, highly selective and orally-bioavailable MMP-13 inhibitors have been identified based upon a (pyridin-4-yl)-2H-tetrazole scaffold. Co-crystal structure analysis revealed that the inhibitors bind at the S(1)(') active site pocket and are not ligands for the catalytic zinc atom. Compound 29b demonstrated reduction of cartilage degradation biomarker (TIINE) levels associated with cartilage protection in a preclinical rat osteoarthritis model. PMID- 20005098 TI - One pot synthesis and SAR of some novel 3-substituted 5,6-diphenyl-1,2,4 triazines as antifungal agents. AB - An improved protocol for the synthesis of a novel series of 1,2,4-triazines possessing 1,2,3-triazole and piperidine ring using 1-(1-substituted piperidin-4 yl)-1H-1,2,3-triazole-4-carbohydrazide, benzil, ammonium acetate and ZrOCl(2).8H(2)O as a catalyst in ethanol-water has been presented. The yields obtained are in the range of 87-94%. All the synthesized compounds (4a-4l) are novel and were evaluated for their in vitro antifungal activity. SAR for the series has been developed by comparing their MIC values with miconazole and fluconazole. Based on activity data SAR for the series has been developed. Compound 4c from the series was equipotent to miconazole against Candida albicans (MIC-25), Aspergillus niger (MIC-12.5) and Cryptococcus neoformans (MIC-25). Compound 4d was equipotent with miconazole against all tested organisms except Cryptococcus neoformans. Also compound 4i was equipotent with miconazole against C. albicans, A. niger and Fusarium oxysporum. PMID- 20005099 TI - Fluorinated piperidine acetic acids as gamma-secretase modulators. AB - We report herein a novel series of difluoropiperidine acetic acids as modulators of gamma-secretase. Synthesis of 2-aryl-3,3-difluoropiperidine analogs was facilitated by a unique and selective beta-difluorination with Selectfluor. Compounds 1f and 2c were selected for in vivo assessment and demonstrated selective lowering of Abeta42 in a genetically engineered mouse model of APP processing. Moreover, in a 7-day safety study, rats treated orally with compound 1f (250mg/kg per day, AUC(0-24)=2100microMh) did not exhibit Notch-related effects. PMID- 20005100 TI - Synthesis and SAR of sulfonyl- and phosphoryl amidine compounds as anti resorptive agents. AB - Sulfonyl amidines (1) and phosphoryl amidines (2), which were efficiently synthesized via a Cu-catalyzed one pot reaction, showed potent anti-bone resorptive activity in vitro. Structure activity relationship studies led to the identification of numerous osteoclast differentiation inhibitors. PMID- 20005101 TI - Isoxazolopyridone derivatives as allosteric metabotropic glutamate receptor 7 antagonists. AB - This Letter describes the synthesis and evaluation of mGluR7 antagonists in the isoxazolopyridone series. In the course of modification in this class, novel solid support synthesis of the isoxazolopyridone scaffold was developed. Subsequent chemical modification led to the identification of several potent derivatives with improved physicochemical properties compared to a hit compound 1. Among these, 2 showed good oral bioavailability and brain penetrability, suggesting that 2 may be useful for in vivo study to elucidate the role of mGluR7. PMID- 20005102 TI - 2,3,5-Trisubstituted pyridines as selective AKT inhibitors. Part II: Improved drug-like properties and kinase selectivity from azaindazoles. AB - A novel series of AKT inhibitors containing 2,3,5-trisubstituted pyridines with novel azaindazoles as hinge binding elements are described. Among these, the 4,7 diazaindazole compound 2c has improved drug-like properties and kinase selectivity than those of indazole 1, and displays greater than 80% inhibition of GSK3beta phosphorylation in a BT474 tumor xenograft model in mice. PMID- 20005103 TI - Synthesis and in vivo evaluation of bicyclic gababutins. AB - Synthesis of a number of bicyclic five-membered ring derivatives of gabapentin led to the identification of two compounds, (-)-(11A) and (20A) which both had an excellent level of potency against alpha(2)delta and were profiled in an in vivo model of neuropathic pain. PMID- 20005104 TI - The first synthetic agonists of FFA2: Discovery and SAR of phenylacetamides as allosteric modulators. AB - Free fatty acid receptor 2 (FFA2) is a G-protein coupled receptor for which only short-chain fatty acids (SCFAs) have been reported as endogenous ligands. We describe the discovery and optimization of phenylacetamides as allosteric agonists of FFA2. These novel ligands can suppress adipocyte lipolysis in vitro and reduce plasma FFA levels in vivo, suggesting that these allosteric modulators can serve as pharmacological tools for exploring the potential function of FFA2 in various disease conditions. PMID- 20005105 TI - Identification of microRNA processing determinants by random mutagenesis of Arabidopsis MIR172a precursor. AB - MicroRNAs (miRNAs) are widespread posttranscriptional regulators of gene expression. They are processed from longer primary transcripts that contain foldback structures (reviewed in). In animals, a complex formed by Drosha and DGCR8/Pasha recognizes the transition between the single-stranded RNA sequences and the stem loop to produce the first cleavage step in miRNA biogenesis. Whereas animal precursors are of uniform size and shape, their plant counterparts comprise a collection of variable stem loops, and little is known about the structural clues recognized during their processing. Here, we designed an unbiased approach based on the random mutagenesis of the MIR172a precursor to study miRNA processing in plants. Randomly mutated precursors were overexpressed in Arabidopsis, and their activity was determined in vivo. We gathered sequence data from these transgenes and used it to build a MIR172a precursor map highlighting relevant and neutral positions for its processing. A 15 nucleotide stem segment below the miRNA/miRNA(*) duplex was essential for MIR172a processing. In contrast, mutations in the terminal-loop region were mostly neutral, yet a loop was required for miR172 biogenesis. The results could be extended to other precursors, suggesting the existence of common features in at least part of the plant precursors. PMID- 20005106 TI - Preferential ethanol consumption in Drosophila models features of addiction. AB - Alcohol addiction is a common affliction with a strong genetic component [1]. Although mammalian studies have provided significant insight into the molecular mechanisms underlying ethanol consumption [2], other organisms such as Drosophila melanogaster are better suited for unbiased, forward genetic approaches to identify novel genes. Behavioral responses to ethanol, such as hyperactivity, sedation, and tolerance, are conserved between flies and mammals [3, 4], as are the underlying molecular pathways [5-9]. However, few studies have investigated ethanol self-administration in flies [10]. Here we characterize ethanol consumption and preference in Drosophila. Flies prefer to consume ethanol containing food over regular food, and this preference increases over time. Flies are attracted to the smell of ethanol, which partially mediates ethanol preference, but are averse to its taste. Preference for consuming ethanol is not entirely explained by attraction to either its sensory or caloric properties. We demonstrate that flies can exhibit features of alcohol addiction. First, flies self-administer ethanol to pharmacologically relevant concentrations. Second, flies will overcome an aversive stimulus in order to consume ethanol. Third, flies rapidly return to high levels of ethanol consumption after a period of imposed abstinence. Thus, ethanol preference in Drosophila provides a new model for studying aspects of addiction. PMID- 20005107 TI - A nonprocessive class V myosin drives cargo processively when a kinesin- related protein is a passenger. AB - During secretory events, kinesin transports cargo along microtubules and then shifts control to myosin V for delivery on actin filaments to the cell membrane [1]. When kinesin and myosin V are present on the same cargo, kinesin interacts electrostatically with actin to enhance myosin V-based transport in vitro [2]. The relevance of this observation within the cell was questioned. In budding yeast, overexpression of a kinesin-family protein (Smy1p) suppressed a transport defect in a strain with a mutant class V myosin (Myo2p) [3]. We postulate that this is a cellular manifestation of the in vitro observation. We demonstrate that Smy1p binds electrostatically to actin bundles. Although a single Myo2p cannot transport cargo along actin bundles, addition of Smy1p causes the complex to undergo long-range, continuous movement. We propose that the kinesin-family protein acts as a tether that prevents cargo dissociation from actin, allowing the myosin to take many steps before dissociating. We demonstrate that both the tether and the motor reside on moving secretory vesicles in yeast cells, a necessary feature for this mechanism to apply in vivo. The presence of both kinesin and myosin on the same cargo may be a general mechanism to enhance cellular transport in yeast and higher organisms. PMID- 20005109 TI - Repeated co-options of exoskeleton formation during wing-to-elytron evolution in beetles. AB - BACKGROUND: The vast diversity in morphology of insect wings provides an excellent model to study morphological evolution. The best-described wing modification is the specification of halteres in Drosophila by a Hox-dependent mechanism, in which a Hox gene affects the expression of genes important for wing development to modify the resulting structure. We have previously shown that highly modified beetle elytra are Hox-free structures despite their divergent morphology, suggesting another mode of evolutionary modification. RESULTS: To understand how elytra have evolved without Hox input, we have analyzed wing development in a coleopteran, Tribolium castaneum. Based on Drosophila mutant phenotypes, we first hypothesized that changes in the wing gene network might have contributed to elytral evolution. However, we found that the wing gene network defined in Drosophila is largely conserved in Tribolium and is also used to pattern the elytra. Instead, we found evidence that the exoskeleton formation has been co-opted downstream of the conserved wing gene network multiple times. We also show evidence that one of these co-options happened prior to the others, suggesting that repeated co-options may have strengthened an advantageous trait. In addition, we found that the Tribolium apterous genes are not only essential for exoskeletalization of the elytra but also are required for the proper identity of the hindwing-an unexpected role that we find to be conserved in Drosophila. CONCLUSIONS: Our findings suggest that elytral evolution has been achieved by co-opting a beneficial trait several times while conserving the main framework of wing patterning genes. PMID- 20005108 TI - RalA-exocyst complex regulates integrin-dependent membrane raft exocytosis and growth signaling. AB - Anchorage dependence of cell growth is a key metastasis-suppression mechanism that is mediated by effects of integrins on growth signaling pathways. The small GTPase RalA is activated in metastatic cancers through multiple mechanisms and specifically induces anchorage independence. Loss of integrin-mediated adhesion triggers caveolin-dependent internalization of cholesterol- and sphingolipid-rich lipid raft microdomains to the recycling endosomes; these domains serve as platforms for many signaling pathways, and their clearance from the plasma membrane (PM) after cell detachment suppresses growth signaling. Conversely, readhesion triggers their return to the PM and restores growth signaling. Activation of Arf6 by integrins mediates exit of raft markers from the recycling endosomes but is not sufficient for return to the PM. We now show that RalA but not RalB mediates integrin-dependent membrane raft exocytosis through the exocyst complex. Constitutively active RalA restores membrane raft targeting to promote anchorage-independent growth signaling. Ras-transformed pancreatic cancer cells also show RalA-dependent constitutive PM raft targeting. These results identify RalA as a key determinant of integrin-dependent membrane raft trafficking and regulation of growth signaling. They therefore define a mechanism by which RalA regulates anchorage dependence and provide a new link between integrin signaling and cancer. PMID- 20005110 TI - mRNA export from mammalian cell nuclei is dependent on GANP. AB - Bulk nuclear export of messenger ribonucleoproteins (mRNPs) through nuclear pore complexes (NPCs) is mediated by NXF1. It binds mRNPs through adaptor proteins such as ALY and SR splicing factors and mediates translocation through the central NPC transport channel via transient interactions with FG nucleoporins. Here, we show that mammalian cells require GANP (germinal center-associated nuclear protein) for efficient mRNP nuclear export and for efficient recruitment of NXF1 to NPCs. Separate regions of GANP show local homology to FG nucleoporins, the yeast mRNA export factor Sac3p, and the mammalian MCM3 acetyltransferase. GANP interacts with both NXF1 and NPCs and partitions between NPCs and the nuclear interior. GANP depletion inhibits mRNA export, with retention of mRNPs and NXF1 in punctate foci within the nucleus. The GANP N-terminal region that contains FG motifs interacts with the NXF1 FG-binding domain. Overexpression of this GANP fragment leads to nuclear accumulation of both poly(A)(+)RNA and NXF1. Treatment with transcription inhibitors redistributes GANP from NPCs into foci throughout the nucleus. These results establish GANP as an integral component of the mammalian mRNA export machinery and suggest a model whereby GANP facilitates the transfer of NXF1-containing mRNPs to NPCs. PMID- 20005111 TI - Bioengineering research in tissue viability. PMID- 20005112 TI - Validity and reliability of the Nintendo Wii Balance Board for assessment of standing balance. AB - Impaired standing balance has a detrimental effect on a person's functional ability and increases their risk of falling. There is currently no validated system which can precisely quantify center of pressure (COP), an important component of standing balance, while being inexpensive, portable and widely available. The Wii Balance Board (WBB) fits these criteria, and we examined its validity in comparison with the 'gold standard'-a laboratory-grade force platform (FP). Thirty subjects without lower limb pathology performed a combination of single and double leg standing balance tests with eyes open or closed on two separate occasions. Data from the WBB were acquired using a laptop computer. The test-retest reliability for COP path length for each of the testing devices, including a comparison of the WBB and FP data, was examined using intraclass correlation coefficients (ICC), Bland-Altman plots (BAP) and minimum detectable change (MDC). Both devices exhibited good to excellent COP path length test retest reliability within-device (ICC=0.66-0.94) and between-device (ICC=0.77 0.89) on all testing protocols. Examination of the BAP revealed no relationship between the difference and the mean in any test, however the MDC values for the WBB did exceed those of the FP in three of the four tests. These findings suggest that the WBB is a valid tool for assessing standing balance. Given that the WBB is portable, widely available and a fraction of the cost of a FP, it could provide the average clinician with a standing balance assessment tool suitable for the clinical setting. PMID- 20005113 TI - Center of mass position relative to the ankle during walking: a clinically feasible detection method for gait imbalance. AB - This study examined the effectiveness of using the center of mass (COM)-Ankle inclination angles to identify elderly individuals with gait imbalance. Whole body motions of 12 elderly patients with gait imbalance and 12 healthy elderly controls were captured with an 8-camera motion analysis system. Inclination angles between the line formed between the COM and lateral ankle marker or by the COM and center of pressure (COP) and their respective vertical lines were computed for each frame during the single stance phase of gait. Similar to the results for COM-COP angles, the results showed that COM-Ankle inclination angles during the single stance phase could successfully distinguish elderly individuals with gait imbalance from healthy elderly adults. Examination of the COM-Ankle inclination angles could be an alternative assessment to clinical populations who possess inconsistent gait that hinder the calculation of the COM-COP inclination angles. PMID- 20005114 TI - Exploring innate glycopeptide resistance mechanisms in Staphylococcus aureus. PMID- 20005115 TI - Probes for narcotic receptor mediated phenomena. 40. N-substituted cis-4a-ethyl 1,2,3,4,4a,9a-hexahydrobenzofuro[2,3-c]pyridin-8-ols. AB - A series of N-substituted rac-cis-4a-ethyl-1,2,3,4,4a,9a-hexahydrobenzofuro[2,3 c]pyridin-8-ols have been prepared using a simple synthetic route previously designed for synthesis of related cis-2-methyl-4a-alkyl-1,2,3,4,4a,9a hexahydrobenzofuro[2,3-c]pyridin-6-ols. The new phenolic compounds, where the aromatic hydroxy moiety is situated ortho to the oxygen atom in the oxide-bridged ring, do not interact as well as the pyridin-6-ols with opioid receptors. The N para-fluorophenethyl derivative had the highest mu-opioid receptor affinity of the examined compounds (K(i)=0.35 microM). PMID- 20005116 TI - Synthesis, biological evaluation and molecular docking studies of amide-coupled benzoic nitrogen mustard derivatives as potential antitumor agents. AB - A series of amide-coupled benzoic nitrogen mustard derivatives as potential EGFR and HER-2 kinase inhibitors were synthesized and reported for the first time. Some of them exhibited significant EGFR and HER-2 inhibitory activity. Of all the studied compounds, compounds 5b and 5t exhibited the most potent inhibitory activity, which was comparable to the positive control erlotinib. Docking simulation was performed to position compounds 5b and 5t into the EGFR active site to determine the probable binding model. Antiproliferative assay results indicated that some of the benzoic nitrogen mustard derivatives possessed high antiproliferative activity against MCF-7. In particular, compounds 5b and 5t with potent inhibitory activity in tumor growth inhibition may function as potential antitumor agents. PMID- 20005117 TI - Design, synthesis and biological evaluation of 6-(benzyloxy)-4-methylquinolin 2(1H)-one derivatives as PDE3 inhibitors. AB - Selective PDE3 (phosphodiesterase 3) inhibitors improve cardiac contractility and may be used in congestive heart failure. However, their proarrhythmic potential is the most important side effect. In this work ten new synthetic compounds (3 [(4-methyl-2-oxo-1,2-dihydro-6-quinolinyl)oxy]methylbenzamide analogs: 4a-j) were designed, synthesized and tested for the inhibitory activity against human PDE3A and PDE3B. The strategy of the design was based on the structure of vesnarinone (a selective PDE3 inhibitor) and its docking analysis results. The synthetic compounds showed better PDE3 inhibitory activity in comparison with vesnarinone. Using docking analysis, a common binding model of each compound toward PDE3 was suggested. In the next step the potential cardiotonic activity of the best PDE3A inhibitors (4b, IC(50)=0.43+/-0.04microM) was evaluated by using the spontaneously beating atria model. In the experiment, atrium of reserpine-treated rat was isolated and the contractile and chronotropic effects of the synthetic compound were assessed. That was carried out in comparison with vesnarinone. The best pharmacological profile was obtained for the compound 4b, which displayed selectivity for increasing the force of contraction (46+/-3% change over the control) rather than the frequency rate (16+/-4% change over the control) at 100microM. PMID- 20005118 TI - Acute and subchronic exposure of diplopods to substrate containing sewage mud: Tissular responses of the midgut. AB - Several discussions regarding the agricultural use of sewage mud have occurred; however, its use has been questioned due to the risks of soil contamination mainly by metals. Diplopods are saprophagous invertebrates, agile in colonizing several layers of soil. Due to the habit of this group, several researchers have proposed their use in ecotoxicological analyses. This study aimed to expose these invertebrates to substrate containing sewage mud of a STS (Sewage Treatment Station) from Sao Paulo State, Brazil and analyzed its toxic potential by morphological analysis of the midgut of the diplopod Rhinocricus padbergi, region of the digestive tube where digestion and nutrient absorption occur. The animals were exposed in mud concentrations at 1%, 10% and 50% mixed with soil from the collection site for periods of 7 (acute exposure), 15 (intermediary exposure) and 90 (subchronic exposure) days in each treatment. The animals from the control group did not present alterations at all the exposition times. As an acute response (7 days), it was observed in the animals exposed to mud at 1% and 10% an increase in the epithelial renovation and in the liberation of secretory vesicles of glycoprotein content. In the animals exposed to mud at 50%, the main alteration observed was the increase in the number of cytoplasmatic granules in the fat body cells; the epithelium seemed to be in reorganization process in all the observed individuals, suggesting an intense epithelial renovation. In the intermediary period of exposure (15 days), in the animals exposed to mud at 1% it was observed an increase in the number of haemocytes among the cells of the fat body, distributed both separately and grouped; the animals exposed to mud at 10% presented the same evidence of epithelial renovation observed in the animals exposed for 7 days at a concentration of 50% of mud; it was also observed an increase in the number of haemocytes, which reacted strongly to the PAS technique. Cytoplasmatic granules in the hepatic cells, fat body cells and haemocytes were also observed in great number, being that great part presented calcium in their constitution. In the animals exposed to mud at 50%, the alteration observed was the presence of numerous secretory vesicles of the apocrine type through all the epithelia of vacuolated aspect; hepatic cells with cytoplasmatic granules were also observed. As subchronic response (90 days), it was observed that all the fat body cells were completely taken by cytoplasmatic granules, into which it was, detected the presence of calcium; such granules were also observed in the apical region of the principal epithelial cells. These results suggest the existence of toxic agents in the analyzed mud sample, whose different concentrations trigger responses that occur in different manners, intensities and velocities that might prevent the entrance and action of toxic agents in the organism of the studied invertebrate. PMID- 20005119 TI - Total efficiency of GE detectors--dead layer signal effect. AB - Two types of detectors (GC 4018 and BE 5030) were compared regarding the signal from the outer dead layer of the detector. Gamma-spectra of Am-241 were acquired with various delays against the starting signal from an alpha LS probe. Coincidence spectra from both detectors were different. The dead-layer signal does not increase the full-energy peak efficiency but it increases the total efficiency, as it was demonstrated by the reconstruction of Am-241 spectra for both detectors. PMID- 20005120 TI - Development of a test for evaluation of taste perception after tongue reduction. AB - When performing a tongue reduction a frequently asked question is how operation will influence taste of the patient. Different kinds of taste tests are designed, most of these being non-specific ways to determine taste sensation in which high concentration of taste solutions are used to detect if a person is able to taste. To be able to judge the influence of tongue reduction on taste we wanted to develop a validated test that could be used in early childhood. No specific tasting areas were found. This test can be used to evaluate tongue reduction procedures. PMID- 20005121 TI - "Mandibular distraction osteogenesis for severe airway obstruction in Robin Sequence. Case report". AB - Mandibular distraction osteogenesis (MDO) has been increasingly gaining interest over the last decade as a treatment alternative for the challenging airway management in infants with the Robin Sequence (RS). This paper is a case report of a male child diagnosed with RS, suffering from life-threatening airway obstruction and feeding difficulties, treated with tracheostomy and gastrostomy since infancy. After evaluation of the patient by a multidisciplinary team of specialists, MDO performed as soon as possible, was considered the optimal treatment, not only to address the severe micrognathia but also to allow early tracheal decannulation. As the lack of space intraorally contraindicated the use of internal distractors, they were placed externally bilaterally. The patient was successfully decannulated 3 weeks postoperatively and the gastrostomy was removed 1 month postoperatively. The mandibular expansion exceeded 20mm bilaterally and the maxilla-mandible discrepancy was fully corrected. There were no complications related to device placement, activation or removal. Follow-up clinical and radiographic examinations of the patient 1 year after the removal of the distractors revealed improved mandibular projection and continued mandibular growth. No significant scarring occurred at the surgical site and the patient has normal respiratory and feeding function. PMID- 20005122 TI - In the name of the Muse. PMID- 20005123 TI - Comprehensive geriatric assessment can predict complications in elderly patients after elective surgery for colorectal cancer: a prospective observational cohort study. AB - OBJECTIVE: To examine the association between the outcomes of a pre-operative comprehensive geriatric assessment (CGA) and the risk of severe post-operative complications in elderly patients electively operated for colorectal cancer. METHODS: One hundred seventy-eight consecutive patients >= 70 years electively operated for all stages of colorectal cancer were prospectively examined. A pre operative CGA was performed, and patients were categorized as fit, intermediate, or frail. The main outcome measure was severe complications within 30 days of surgery. RESULTS: Twenty-one patients (12%) were categorized as fit, 81 (46%) as intermediate, and 76 (43%) as frail. Eighty-three patients experienced severe complications, including three deaths; 7/21 (33%) of fit patients, 29/81 (36%) of intermediate patients and 47/76 (62%) of frail patients (p=0.002). Increasing age and ASA classification were not associated with complications in this series. CONCLUSION: CGA can identify frail patients who have a significantly increased risk of severe complications after elective surgery for colorectal cancer. PMID- 20005124 TI - How Hydra senses and destroys microbes. AB - Molecular genetic evidence has revealed that the basic templates of innate immune sensors were laid down in ancient animals such as the cnidarian Hydra. Important functions of Hydra's innate immune sensors and effectors include not only protection against pathogens but also controlling tissue-microbiota homeostasis. The deep evolutionary connections imply that invertebrate and mammalian immune pathways have evolved from a reduced number of common ancestral building blocks to their present configurations. PMID- 20005126 TI - Position accuracy and electromyographic responses during head reposition in young adults with chronic neck pain. AB - The head reposition test is traditionally used to examine the proprioceptive sensitivity of the neck. The aim of this study was to investigate the position accuracy and corresponding cervical electromyographic (EMG) responses of the neck pain subjects during sagittal head-to-neutral tasks. Twelve young subjects with chronic neck pain and twelve young healthy subjects were recruited. The position accuracy was measured by the constant error, variable error, and root mean square error of joint angles during head-to-neutral tasks in flexion and extension directions. Surface EMG of neck flexors and extensors were analyzed by the voluntary response index, including the similarity index (SI) and electromyographic magnitude (MAG) of muscle groups. The normalized average integration of EMG activity (NAIEMG) of individual muscle was also calculated. The results showed: (1) significantly larger constant error and root mean square error but similar variable error in patients compared with controls, (2) smaller SI but similar MAG in patients compared with controls, (3) greater synergistic/antagonistic NAIEMG in patients than controls. The findings suggested that young adults with chronic neck pain exhibit proprioceptive dysfunction and altered EMG pattern during voluntary sagittal neck motions. This study provides guidelines which could lead to the development of therapeutic exercise programs. PMID- 20005125 TI - Cloning and expression of a region of vesicle associated membrane protein2 (VAMP2) gene and its use as a recombinant peptide substrate for assaying clostridial neurotoxins in contaminated biologicals. AB - An assay for the endopeptidase activities of clostridial neurotoxins in contaminated biotherapeutic products has been developed. Based on a synthetic peptide substrate representing amino acid residues 60-94 of the intracellular vesicle associated membrane protein2 (VAMP2), RT-PCR was used to amplify the VAMP2 sequence. The extended insert was digested with EcoRI and SalI and ligated into pGEX4T-1 vector for construction of the pGEX4T-1/VAMP plasmid for expressing in Escherichia coli a fusion protein linked to glutathione S-transferase (GST). The fusion protein was purified by affinity chromatography and used in an ELISA assay for comparison with the commercially available synthetic VAMP peptide and rabbit polyclonal antiserum. The identity of the immunoreactivity of recombinant VAMP2 protein with the chemically synthesized peptide was demonstrated by western blot. Our results indicated that recombinant VAMP2 peptide not only reacted with specific polyclonal antibody in a dose-dependent manner, without any remarkable difference observed between the reactivity of the fusion protein and commercial VAMP2 segment peptide, but also cleaved by botulinum neurotoxin type B (BONT/B) after endopeptidase assay. Thus, recombinant VAMP2 could serve as a replacement for VAMP2 synthetic peptide, potentially useful in endopeptidase assays for replacement of the currently used mouse bioassay for clostridial neurotoxins contaminating biotherapeutic products. PMID- 20005127 TI - Test-retest reliability of the soleus H-reflex is affected by joint positions and muscle force levels. AB - The purpose of this study was to determine the test-retest reliability of the soleus (SOL) H-reflex during rest and isometric contractions at 10%, 30%, and 50% of the maximal voluntary force (MVC) at the ankle joint angles of neutral (0 degrees), plantarflexion (20 degrees), and dorsiflexion (-20 degrees) respectively, in a sitting position. Ten healthy participants, with mean age of 24.9+/-5.0 (SD) years, height 168.3+/-8.8 cm, weight 62.7+/-12.3 kg, were tested for the SOL H-reflex (H(max)) on two separate occasions within 7 days. The intraclass correlation coefficient (ICC) for the test-retest of the SOL H-reflex during rest was found to be high at ankle joint angle of neutral (ICC=0.92) and plantarflexion (0.96), and moderate at dorsiflexion (0.75). Inconsistent ICC values (range from 0.62 to 0.97) were found during the submaximal voluntary contractions at the three ankle joint positions. High ICCs were also found in H(max)/M(max) ratio at neutral (0.86), plantarflexion (0.96), and dorsiflexion (0.84) positions. It was concluded that the test-retest reliability of the SOL H reflex was affected by the intensity of voluntary contraction and ankle joint position. The H-reflex demonstrated a higher reliability at the neutral and plantarflexion positions than that at the dorsiflexion position during rest, and a higher reliability at 10% MVC than that at 30% and 50% MVC. PMID- 20005128 TI - Classification of multi muscle activation patterns of osteoarthritis patients during level walking. AB - The study compares the timing and frequency changes of surface EMGs recorded from osteoarthritis patients with previous traumatic ankle injury and normal subjects during level walking. EMG intensity (power) was obtained by a wavelet analysis. There were intensity values for each frequency characterized by the wavelets for every time point. The intensities were compounded into Multi Muscle Patterns (MMP) simultaneously showing the time and spectral aspects of the lower leg muscle activity. The aim of the study was to test the hypothesis that the differences between the group of the MMPs from the affected leg (AFL) and the not affected leg (NAL) allow detecting whether a newly measured MMP results from an AFL or NAL. This hypothesis was tested by a spherical classification procedure yielding the correctly classified MMPs thus indicating the significance of the differences between the MMPs of the AFL and NAL. The hypothesis was supported (not falsified) by the results. Thus there were common features of muscle activity in the AFL of most osteoarthritis patients that allowed detecting whether the MMP of a new patient was of the kind seen in most other osteoarthritis patients. The spectral, timing and intensity factors in the MMP that allowed this classification were visualized in the mean MMPs of the patients and the control group. The comparison revealed where on average the relative timing and spectral differences of the muscle activation of osteoarthritis patients and control subjects occurred. PMID- 20005129 TI - Epidural analgesia improves outcome in cardiac surgery: a meta-analysis of randomized controlled trials. AB - OBJECTIVE: The authors conducted a review of randomized studies to determine whether there were any advantages for clinically relevant outcomes by adding epidural analgesia in patients undergoing cardiac surgery under general anesthesia. DESIGN: Meta-analysis. SETTING: Hospitals. PARTICIPANTS: A total of 2366 patients from 33 randomized trials. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: DATA SOURCES AND STUDY SELECTION: PubMed, BioMedCentral, CENTRAL, EMBASE, Cochrane Central Register of Controlled Trials, and conference proceedings were searched (updated January 2008) for randomized trials that compared general anesthesia with an anesthetic plan including general anesthesia and epidural analgesia in cardiac surgery. Two independent reviewers appraised study quality, with divergences resolved by consensus. Overall analysis showed that epidural analgesia reduced the risk of the composite endpoint mortality and myocardial infarction (30/1125 [2.7%] in the epidural group v 64/1241 [5.2%] in the control arm, odds ratio [OR] = 0.61 [0.40-0.95], p = 0.03 number needed to treat [NNT] = 40), the risk of acute renal failure (35/590 [5.9%] in the epidural group v 54/618 [8.7%] in the control arm, OR = 0.56 [0.34-0.93], p = 0.02, NNT = 36), and the time of mechanical ventilation (weighted mean differences = -2.48 hours [-2.64, -2.32], p < 0.001). CONCLUSIONS: This analysis suggested that epidural analgesia on top of general anesthesia reduced the incidence of perioperative acute renal failure, the time on mechanical ventilation, and the composite endpoint of mortality and myocardial infarction in patients undergoing cardiac surgery. PMID- 20005130 TI - Increased alveolar damage after mechanical ventilation in a porcine model of thoracic surgery. AB - OBJECTIVE: Mechanical stress during one-lung ventilation (OLV) results in lung injury. This study compared the effects of mechanical ventilation, OLV, and surgical manipulation on diffuse alveolar damage (DAD) after application of different anesthetic regimens. DESIGN: Prospective, randomized, controlled, blinded animal experiment. SETTING: University hospital. OBJECTS: Twenty-one piglets. INTERVENTIONS: Animals (27.5 kg) were randomized into 4 groups: spontaneous breathing (SB, n = 3), two-lung ventilation (TLV, n = 6), OLV during desflurane (n = 6), and propofol anesthesia (n = 6). SB pigs were killed after the induction of anesthesia. Lung tissue samples were analyzed to obtain reference values for alveolar damage. TLV pigs underwent standard TLV (tidal volumes [V(T)] = 10 mL/kg, F(I)O(2) = 0.40, positive end-expiratory pressure = 5 cmH(2)O). In OLV pigs, after lung separation by a bronchial blocker, OLV (V(T) = 10 mL/kg) and thoracic surgery were performed. After the procedure, the pigs were killed. Lung tissue samples were harvested for histologic examination. Lung injury was quantified by DAD score; sequestration of leukocytes was assessed by the recruitment of CD45(+) cells into the lungs. MAIN RESULTS: TLV resulted in increased DAD scores in both lungs (TLV v SB: 6.9 v 2.7, p < 0.05); the number of CD45(+) cells was not increased (TLV v SB: 8.7 v 5.0 cells per view). OLV and surgical manipulation increased DAD and leukocyte sequestration without differences between the ventilated and manipulated lungs. Leukocyte recruitment was not differently affected by the anesthetic regimen (propofol v desflurane: CD45(+) cells per view: 13.5 v 11.3). CONCLUSIONS: TLV resulted in increased DAD scores in the lungs as compared with SB. OLV and thoracic surgery further increased lung injury and leukocyte recruitment independently of the administration of propofol or desflurane anesthesia. PMID- 20005131 TI - Cardiac index validation using the pressure recording analytic method in unstable patients. AB - OBJECTIVE: The authors investigated the accuracy and precision of the pressure recording analytic method (PRAM) in cardiac index measurement compared with thermodilution in unstable patients, a setting in which minimally invasive monitoring devices often fail. DESIGN: Criterion standard. SETTING: Intensive care unit. PATIENTS: Thirty-two consecutive patients with low cardiac output syndrome treated with an intra-aortic balloon pump and/or high doses of inotropic drugs but without atrial fibrillation were studied after cardiac surgery. INTERVENTIONS: None. Pulmonary and radial artery catheters were already in situ for clinical reasons. MEASUREMENTS AND MAIN RESULTS: Four patients (12.5%) were excluded from the study because of artifacts caused by under- or overdamping of the arterial pressure monitoring system. The authors performed 3 injections of the thermal indicator in 5 minutes through the pulmonary artery catheter. Mean cardiac index values of 12 consecutive beats were considered for the PRAM. A significant correlation was found between the cardiac index assessed by thermodilution and PRAM (r = 0.72, p < 0.001). The mean bias between the 2 techniques was 0.072 +/- 0.41 L/min/m(2) with lower and upper 95% limits of confidence of -0.089 and 0.233 L/min/m(2), respectively. The percentage error was 30%. Sufficient agreement between the two techniques was evidenced by the Bland Altman plot with only two points above the limits of agreement. CONCLUSIONS: This study showed that PRAM, a minimally invasive method for cardiac index assessment, is clinically useful even in unstable patients such as those receiving intra aortic balloon pump and/or ongoing high doses of a inotropic drugs because of a low cardiac output syndrome but without atrial fibrillation. PMID- 20005132 TI - A new plasma-adapted hydroxyethyl starch preparation: in vitro coagulation studies. AB - OBJECTIVE: Preparing hydroxyethyl starch (HES) in a plasma-adapted solution is supposed to improve safety with regard to coagulation. The influence of a new plasma-adapted HES preparation on coagulation was studied. DESIGN: Operator blinded, randomized study. SETTING: Laboratory in vitro study. PARTICIPANTS: Fifteen healthy young men scheduled for blood donation. INTERVENTIONS: Blood was diluted by 10%, 30%, and 50% using either a plasma-adapted or nonplasma-adapted (prepared in saline solution) potato-derived 6% HES 130/0.42. Only the composition of the solvent of the 2 HES preparations was different. MEASUREMENTS AND MAIN RESULTS: Rotation thromboelastometry (ROTEM; Pentapharm, Munich, Germany) was used to assess changes in coagulation; whole blood aggregometry with 3 inducers was used to assess effects of dilution with HES on platelet function. Clotting time (CT) and clot formation time were significantly prolonged by 30% and 50% dilution, showing significantly longer times in the non-plasma-adapted than in the plasma-adapted HES group (eg, intrinsic CT at the 30% dilution level: plasma-adapted HES 228 +/- 26 seconds [within normal range] v 269 +/- 29 seconds in the nonplasma-adapted HES group). Clot strengthening and clot firmness were significantly reduced by the non-plasma-adapted HES at the 30% dilution level. Platelet aggregation was significantly more reduced by the non-plasma-adapted HES at the 30% and 50% dilution levels. CONCLUSIONS: Dilution with the nonplasma adapted HES 130/0.42 was associated with more negative effects on thromboelastometry and platelet aggregation than the same HES 130/0.4 dissolved in a plasma-adapted solution. The benefits of using a plasma-adapted modern HES preparation on blood loss and use of blood/blood products in cardiac surgery need to be studied. PMID- 20005133 TI - Effects of loss aversion on post-decision wagering: implications for measures of awareness. AB - Wagering contingent on a previous decision, or post-decision wagering, has recently been proposed to measure conscious awareness. Whilst intuitively appealing, it remains unclear whether economic context interacts with subjective confidence and how such interactions might impact on the measurement of awareness. Here we propose a signal detection model which predicts that advantageous wagers placed on the identity of preceding stimuli are affected by loss aversion, despite stimulus visibility remaining constant. This pattern of predicted results was evident in a psychophysical task where we independently manipulated perceptual and economic factors. Changes in wagering behaviour induced by changes in wager size were largely driven by changes in criterion, consistent with the model. However, for near-threshold stimuli, a reduction in wagering efficiency was also evident, consistent with an apparent but potentially illusory decrease in awareness of the stimulus. These findings challenge an assertion that post-decision wagering provides a direct index of subjective awareness. PMID- 20005134 TI - T cell receptor CDR3 sequence but not recognition characteristics distinguish autoreactive effector and Foxp3(+) regulatory T cells. AB - The source, specificity, and plasticity of the forkhead box transcription factor 3 (Foxp3)(+) regulatory T (Treg) and conventional T (Tconv) cell populations active at sites of autoimmune pathology are not well characterized. To evaluate this, we combined global repertoire analyses and functional assessments of isolated T cell receptors (TCR) from TCRalpha retrogenic mice with autoimmune encephalomyelitis. Treg and Tconv cell TCR repertoires were distinct, and autoantigen-specific Treg and Tconv cells were enriched in diseased tissue. Autoantigen sensitivity and fine specificity of these cells intersected, implying that differences in responsiveness were not responsible for lineage specification. Notably, autoreactive Treg and Tconv cells could be fully distinguished by an acidic versus aliphatic variation at a single TCR CDR3 residue. Our results imply that ontogenically distinct Treg and Tconv cell repertoires with convergent specificities for autoantigen respond during autoimmunity and argue against more than limited plasticity between Treg and Tconv cells during autoimmune inflammation. PMID- 20005135 TI - MicroRNA-132 potentiates cholinergic anti-inflammatory signaling by targeting acetylcholinesterase. AB - MicroRNAs (miRNAs) contribute to both neuronal and immune cell fate, but their involvement in intertissue communication remained unexplored. The brain, via vagal secretion of acetylcholine (ACh), suppresses peripheral inflammation by intercepting cytokine production; therefore, we predicted that microRNAs targeting acetylcholinesterase (AChE) can attenuate inflammation. Here, we report that inflammatory stimuli induced leukocyte overexpression of the AChE-targeting miR-132. Injected locked nucleic acid (LNA)-modified anti-miR-132 oligonucleotide depleted miR-132 amounts while elevating AChE in mouse circulation and tissues. In transfected cells, a mutated 3'UTR miR-132 binding site increased AChE mRNA expression, whereas cells infected with a lentivirus expressing pre-miR-132 showed suppressed AChE. Transgenic mice overexpressing 3'UTR null AChE showed excessive inflammatory mediators and impaired cholinergic anti-inflammatory regulation, in spite of substantial miR-132 upregulation in brain and bone marrow. Our findings identify the AChE mRNA-targeting miR-132 as a functional regulator of the brain-to-body resolution of inflammation, opening avenues for study and therapeutic manipulations of the neuro-immune dialog. PMID- 20005136 TI - Chemokine-induced Zap70 kinase-mediated dissociation of the Vav1-talin complex activates alpha4beta1 integrin for T cell adhesion. AB - Lymphocyte integrins mediate cell arrest on endothelium during immune surveillance after activation by chemokine-stimulated inside-out signals. Here we show that a Vav1-talin complex in T cells is a key target for chemokine-triggered inside-out signaling leading to integrin alpha4beta1 activation. Thus, Vav1 dissociation from talin was required to generate high-affinity alpha4beta1 conformations. Assembly of the Vav1-talin complex required PtdIns(4,5)P(2), which was provided by talin-bound phosphatidylinositol phosphate kinase Igamma. Chemokine-promoted Vav1 dissociation from talin followed an initial increase in talin binding to alpha4beta1. This process was dependent on ZAP-70, which binds to and phosphorylates Vav1 in the complex, leading to further alpha4beta1 activation and cell adhesion strengthening. Moreover, Vav1-talin dissociation was needed for Rac1 activation, thus indicating that alpha4beta1 and Rac1 activation can be coupled by chemokine-stimulated ZAP-70 function. Our data suggest that Vav1 might function as a repressive adaptor of talin that must dissociate from alpha4beta1-talin complexes for efficient integrin activation. PMID- 20005137 TI - Perinatal lethal form of Gaucher disease. Clinical and molecular characterization of a Greek case. PMID- 20005138 TI - Polarization encoding as a novel approach to MRI. AB - In magnetic resonance imaging (MRI), there have been three basic techniques to encode the spatial origin of the signals, i.e. Fourier and radio frequency encoding and the use of sensitivity information of sensor arrays. In this paper, we introduce a new encoding method, which we call polarization encoding. The method utilizes sets of polarizing fields with various spatial profiles; it is tailored for MRI at ultra-low fields (ULF MRI). In ULF MRI, signals from a prepolarized sample are typically detected using an array of SQUID (superconducting quantum interference device) sensors at microtesla fields. The prepolarization is achieved with a field of the order of 10-100mT preceding the signal acquisition. In polarization encoding, the prepolarizing field is varied in a way to gain additional information about the sample. The method may also prove useful for modalities that in the absence of any precession aim to image the DC magnetization profile of a sample. PMID- 20005139 TI - (1)H MRS detection of glycine residue of reduced glutathione in vivo. AB - Glutathione (GSH) is a powerful antioxidant found inside different kinds of cells, including those of the central nervous system. Detection of GSH in the human brain using (1)H MR spectroscopy is hindered by low concentration and spectral overlap with other metabolites. Previous MRS methods focused mainly on the detection of the cysteine residue (GSH-Cys) via editing schemes. This study focuses on the detection of the glycine residue (GSH-Gly), which is overlapped by glutamate and glutamine (Glx) under physiological pH and temperature. The first goal of the study was to obtain the spectral parameters for characterization of the GSH-Gly signal under physiological conditions. The second goal was to investigate a new method of separating GSH-Gly from Glx in vivo. The characterization of the signal was carried out by utilization of numerical simulations as well as experiments over a wide range of magnetic fields (4.0 14T). The proposed separation scheme utilizes J-difference editing to quantify the Glx contribution to separate it from the GSH-Gly signal. The presented method retains 100% of the GSH-Gly signal. The overall increase in signal to noise ratio of the targeted resonance is calculated to yield a significant SNR improvement compared to previously used methods that target GSH-Cys residue. This allows shorter acquisition times for in vivo human clinical studies. PMID- 20005140 TI - Pleiotropic role of IGF-I in obesity hypoventilation syndrome. AB - CONTEXT: Obesity hypoventilation syndrome (OHS) is defined by the association between obesity and daytime arterial hypercapnia. The syndrome includes in variable proportion impaired diaphragmatic weakness, decreased central ventilatory drive and nearly systematically occurrence of sleep apnea. An increased cardio-vascular risk has been demonstrated compared to normocapnic obesity. IGF-I has a pleiotropic role in metabolism, ventilatory control, muscle function and cardiovascular protection. OBJECTIVES AND DESIGN: We performed a case control study comparing somatotropic axis changes including IGF-I in obese with or without OHS. METHODS: Patients underwent respiratory function tests, CO(2) ventilatory responses, polysomnography and somatotropic axis exploration (GH, IGF-I and IGFBP-3). RESULTS: 15 OHS (BMI: 41+/-5.6 kg/m(2), PaCO(2): 6.13+/ 0.39 kPa, age: 55.6+/-5.9 years) and 15 matched obese without hypercapnia (BMI: 42+/-6.7 kg/m(2), PaCO(2): 5.13+/-0.27 kPa, age: 55.0+/-7.5 years) were compared. IGF-I and IGFBP-3 were significantly lowered in OHS, and negatively correlated with PaCO(2) (r=-0.615; P<0.001 and r=-0.452; P=0.016, respectively). Inspiratory capacity and forced vital capacity reflecting respiratory muscle strength decreased significantly with IGF-I (r=0.408; P=0.038). Triglycerides levels were higher in OHS (1.64+/-0.58 versus 1.13+/-0.56 g/L; P<0.01), and negatively associated with IGF-I (r=-0.418; P=0.027). CONCLUSION: A low IGF-I level is associated with hypercapnia presumably by reducing ventilatory drive and favouring muscle weakness. The relationship between increased triglycerides and low IGF-I may represent one of mechanisms involved in the OHS increased cardio vascular risk. PMID- 20005141 TI - Human testis-specific PDHA2 gene: methylation status of a CpG island in the open reading frame correlates with transcriptional activity. AB - DNA methylation is an important epigenetic modification that has profound roles in gene expression and, in particular, is thought to be crucial for regulation of tissue-specific genes in animal cells. The pivotal E(1)alpha subunit of human pyruvate dehydrogenase complex, an essential and rate-limiting enzyme system in energy metabolism, is encoded by two distinct genes: PDHA1 gene, located on chromosome X is expressed in somatic tissues, whereas PDHA2 gene, located on chromosome 4, is exclusively expressed in spermatogenic cells. The objective of this study is to elucidate the role of DNA methylation as an epigenetic mechanism controlling the regulation of PDHA2 gene expression in human tissues, namely its repression in somatic tissues and its activation in testicular germ cells. Genomic DNA was isolated from human somatic tissues (circulating lymphocytes and gastric cells) and from testis, including isolated fractions of haploid and diploid germ cells. After primer design with appropriate software, it was performed the sodium bisulfite PCR sequencing of the PDHA2 promoter and coding regions. Total RNA of the same tissues was isolated, reverse transcribed and PDHA1and PDHA2 transcripts were amplified with specific primers and analysed by agarose gel electrophoresis. The analysis of the genomic sequence of the PDHA2 gene revealed the presence of 61 CpG sites whose distribution matches the criteria for the presence of two CpG islands. Sequence analysis of both CpG islands upon bisulfite treatment displayed several differences, either between islands or among tissues. In particular, the methylation pattern of one of the CpG islands revealed a perfect correlation with transcriptional activity of the PDHA2 gene either in testis or in somatic tissues. Surprisingly, it is the full demethylation of the CpG island located in the coding region that seems to play a crucial role upon PDHA2 gene transcription in testis. PMID- 20005142 TI - Prevalence of coronavirus. PMID- 20005143 TI - [Management of Merkel cell carcinoma: Role of radiotherapy in elderly patients]. AB - PURPOSE: Merkel cell carcinoma carcinoma (MCC) or primary cutaneous neuroendocrine carcinoma is a rare and aggressive malignancy affecting elderly. Optimal therapeutic strategy has not yet been established in elderly patients. PATIENTS AND METHODS: From March 1996 to March 2007, 29 patients with Merkel cell carcinoma of were treated at the University Hospital of Amiens, France. Adjuvant radiotherapy (RT) was performed for 14 patients (50%) on the tumor bed with margins of 3 to 5cm, an average dose of 46Gy (30-60Gy), by 2Gy per fraction. Ten of them also received RT to the lymph node area at mean dose of 44.3Gy (26-50Gy). Duration of RT was 35 days. A retrospective analysis was conducted to better evaluate survival and prognostic factors. RESULTS: Median overall survival (OS) was 18.9 months (3-122) and the median time to progression (MTP) 5.5 months (1 26). At 5 years, OS for irradiated patients was 47% (IC95: 12-82%) versus 27% (IC95: 5-49%) in cases of surgery alone (p=0.032). The most frequent sites of recurrence were nodal (34.5%), local (24.1%) and metastatic (17.2%). For patients over 70 years, eight (36.5%) were free of disease at last news, 8 (36.5%) had died from cancer and six from other causes (27%). In this subgroup, MTP was 6 months (2-19) and median OS of 19 months (4-87). There was no acute toxicity greater than grade 2. CONCLUSION: Although limited by a retrospective analysis, this report suggests an advantage of postoperative RT for patients with MCC. It combined low toxicity and improvement of survival. Prospective multicenter trials are needed to clarify and validate the optimal strategy. PMID- 20005144 TI - Estimation of stature from facial measurements in northwest Indians. AB - Estimation of stature is one of the important component in identification of human remains in forensic anthropology. The present investigation attempts to estimate stature from seven facial measurements of 300 (173 males and 127 females) healthy subjects between the ages of 18-70years from Northwest India. Height of all the subjects was measured and facial measurements were taken. Data was subjected to statistical analysis like mean, standard deviation, multiplication factors, Karl Pearson's correlation coefficient (r), linear and multiple regression analyses using statistical package for social sciences (SPSS). The average height of the subjects was in the range of 154.3-178.3cm in males and 155.1-168.4cm in females. Estimated stature calculated by regression analysis of seven facial measurements was almost similar to mean actual stature in both males and females and the difference by using multiplication factors was found to be greater. Standard error of estimation (SEE) computed both by linear and multiple regression analyses was found to be low for the two sexes. Thus we can conclude that regression equations generated from facial measurements can be a supplementary approach for the estimation of stature when extremities are not available. PMID- 20005145 TI - Ultrasound and microwave assisted synthesis of high loading Fe-supported Fischer Tropsch catalysts. AB - Supported iron-based Fischer-Tropsch (FT) catalysts with high loading of active metal have been prepared using both traditional and innovative methods. In these latter the impregnation of silica support has been performed by adding a step involving an ultrasound (US) or a microwave (MW) treatment to improve the metal deposition and to increase the catalyst activity. FT results have indicated the catalysts prepared by US as the most efficient, particularly when sonication is performed in argon atmosphere. MW prepared samples have given results very similar to those obtained with the traditional method. In order to explain the different catalytic activity, all the samples have been characterized by BET, TPR, SEM, TEM, XRD and micro-Raman analyses. PMID- 20005146 TI - Planned stopping in people with Parkinson. AB - The aim of this study was to quantify gait termination in people with Parkinson (PwP) as the basis for understanding the underlying pathophysiology of stopping difficulties. Fourteen PwP and 14 age- and gender-matched comparisons completed five trials each of four walking tasks: preferred walk, preferred walk with secondary motor task, coming to a planned stop, and planned stop with a secondary motor task. Spatio-temporal data of walks were compared to steady state walking in stopping trials. Results showed that PwP walked with shorter step length, slower speed, yet similar cadence to comparisons. Both groups decreased step length and step speed when performing a secondary task. Neither group showed changes of gait characteristics in steady state walking prior to stopping. For stopping trials, the number of steps, time, and distance taken to stop were compared for PwP and controls. In planned stops PwP used more steps and took a longer time to stop, but both groups stopped within a similar distance. A secondary motor task did not alter stopping distance or number of steps to stop, but stopping time increased in the comparisons. The results indicate that central control mechanisms regulating planned stopping appear to be intact in people with mild to moderate Parkinson. PMID- 20005147 TI - "It's not necessarily the distance on the map...": using place as an analytic tool to elucidate geographic issues central to rural palliative care. AB - Palliative care is intimately connected with place, yet little research has explored these relationships in depth, especially with respect to rural and remote settings. This paper uses multiple dimensions of the concept 'place' as an analytic tool to understand the nature of palliative care provision in a rural region of British Columbia, Canada. We draw upon primary data from formal and informal providers (n = 31) to explore the social and physical place of rural palliative care. We unpack four highly geographic issues raised by participants, namely: (1) distance, (2) location, (3) aesthetics, and (4) sites of care. This analysis reveals a rich and complex experience of rural care-giving long overlooked in palliative care research and policy. PMID- 20005148 TI - Pathways mediating VEGF-independent tumor angiogenesis. AB - FDA approval of several inhibitors of the VEGF pathway has enabled significant advances in the therapy of cancer and neovascular age-related macular degeneration. However, similar to other therapies, inherent/acquired resistance to anti-angiogenic drugs may occur in patients, leading to disease progression. So far the lack of predictive biomarkers has precluded identification of patients most likely to respond to such treatments. Recent suggest that both tumor and non tumor (stromal) cell types are involved in the reduced responsiveness to the treatments. The present review examines the role of tumor- as well as stromal cell-derived pathways involved in tumor growth and in refractoriness to anti-VEGF therapies. PMID- 20005149 TI - CC chemokine ligand 2 (CCL2) promotes prostate cancer tumorigenesis and metastasis. AB - CCL2 is a chemokine known to recruit monocytes and macrophages to sites of inflammation. A growing body of research suggests CCL2 is progressively overexpressed in tumor beds and may play a role in the clinical progression of solid tumors. Cancer cells derived from several solid tumor types demonstrate functional receptors for CCL2, suggesting this chemokine may achieve tumorigenicity through direct effects on malignant cells; however, a variety of normal host cells that co-exist with cancer in the tumor microenvironment also respond to CCL2. These cells include macrophages, osteoclasts, endothelial cells, T-lymphocytes, and myeloid-derived immune suppressor cells (MDSCs). CCL2 mediated interactions between normal and malignant cells in the tumor microenvironment and plays a multi-faceted role in tumor progression. PMID- 20005151 TI - The central regulation of plant physiology by adenylates. AB - There have been many recent developments concerning the metabolic, transport and signalling functions of adenylates in plants, suggesting new roles for these compounds as central regulators of plant physiology. For example, altering the expression levels of enzymes involved in the equilibration, salvaging, synthesis and transport of adenylates leads to perturbations in storage, growth and stress responses, implying a role for adenylates as important signals. Furthermore, sensing of the internal energy status involves SNF1-related kinases, which control the expression and phosphorylation of key metabolic enzymes. ATP also acts as an apoplastic signalling molecule to control cell growth and pathogen responses. These new results could shed light on the emerging question of whether energy homeostasis in plant cells differs from mechanisms found in microbes and mammals. PMID- 20005152 TI - DWI filtering using joint information for DTI and HARDI. AB - The filtering of the Diffusion Weighted Images (DWI) prior to the estimation of the diffusion tensor or other fiber Orientation Distribution Functions (ODF) has been proved to be of paramount importance in the recent literature. More precisely, it has been evidenced that the estimation of the diffusion tensor without a previous filtering stage induces errors which cannot be recovered by further regularization of the tensor field. A number of approaches have been intended to overcome this problem, most of them based on the restoration of each DWI gradient image separately. In this paper we propose a methodology to take advantage of the joint information in the DWI volumes, i.e., the sum of the information given by all DWI channels plus the correlations between them. This way, all the gradient images are filtered together exploiting the first and second order information they share. We adapt this methodology to two filters, namely the Linear Minimum Mean Squared Error (LMMSE) and the Unbiased Non-Local Means (UNLM). These new filters are tested over a wide variety of synthetic and real data showing the convenience of the new approach, especially for High Angular Resolution Diffusion Imaging (HARDI). Among the techniques presented, the joint LMMSE is proved a very attractive approach, since it shows an accuracy similar to UNLM (or even better in some situations) with a much lighter computational load. PMID- 20005153 TI - Dual role of the six-coordinated molybdenum and lead ions in novel of photochromic properties of the molybdenum-lead-borate glasses. AB - Transparent glasses were prepared by conventional melting-quenching method in the xMoO(3).(100-x)[3B(2)O(3).PbO] system where 0 or =31; log-rank p=0.033; HR 0.59, 0.35-1.01), after adjustment for number of positive nodes. The recurrence score by treatment interaction was significant in the first 5 years (p=0.029), with no additional prediction beyond 5 years (p=0.58), although the cumulative benefit remained at 10 years. Results were similar for overall survival and breast-cancer specific survival. INTERPRETATION: The recurrence score is prognostic for tamoxifen-treated patients with positive nodes and predicts significant benefit of CAF in tumours with a high recurrence score. A low recurrence score identifies women who might not benefit from anthracycline-based chemotherapy, despite positive nodes. FUNDING: National Cancer Institute and Genomic Health. PMID- 20005177 TI - First-generation genomic tests for breast cancer treatment. PMID- 20005178 TI - Sexual counselling of cardiac patients: nurses' perception of practice, responsibility and confidence. AB - BACKGROUND: Cardiac patients may experience problems with sexual activity as a result of their disease, medications or anxiety and nurses play an important role in sexual counselling. We studied the practice, responsibility and confidence of cardiac nurses in the sexual counselling of these patients. METHOD: An adapted version of the nurses' survey of sexual counselling of MI patients was administered during a scientific meeting of the Council on Cardiovascular Nursing and Allied Professionals within the European Society of Cardiology. RESULTS: Most of the 157 cardiovascular nurses (87%) who completed the survey felt responsible to discuss sexual concerns with their clients, especially when patients initiated a discussion. However in practice, most respondents rarely addressed sexual issues. The items that nurses reported to counsel patients were closely related to the cardiac disease, symptoms and medications and seldom more sensitive subjects (e.g. foreplay, positions). Nurses estimated that their patients could be upset (67%), embarrassed (72%) or anxious (68%) if they were asked about sexual concerns. One-fifth of the nurses felt they had insufficient knowledge and 40% sometimes hesitated to discuss sexual concerns with clients because they might not know how to answer questions. Additional education on sexuality was significantly related to being more comfortable and active in sexual counselling. CONCLUSION: Although cardiac nurses feel responsible and not anxious discussing patients' sexual concerns, these issues are not often discussed in daily practice. Nurses might need more knowledge and specific practical training in providing information on sexual concerns and sexual counselling to cardiac patients. PMID- 20005179 TI - Sir Denis Browne: contributions to pediatric urology. AB - PURPOSE: Sir Denis Browne, a surgeon at the Hospital for Sick Children, Great Ormond Street, was the first surgeon to confine his practice solely to pediatric surgery. He made numerous important contributions to the field of pediatric surgery in orthopedics, otolaryngology, plastic surgery and urology, and is thus known as the father of modern British pediatric surgery. MATERIAL AND METHODS: We reviewed the original publications, historical records and medical literature on Denis Browne and his contributions to urology. RESULTS: Sir Denis Browne used his anatomical studies to elucidate many of the problems he faced. He classified undescended testis into two groups still recognized today: 'normal variations of development (retractile testis) which only need leaving alone' and 'congenital deformities which need operation'. During orchidopexy he stressed cord mobilization rather than testicular fixation and tension on the blood supply. He was also the first to describe the superficial inguinal pouch, which bears his name, as a common site for the undescended testis. He devised a new operation for hypospadias based on the principle that a buried strip of skin will epithelialize the surrounding tissue into a tube. The simplicity of his technique made hypospadias surgery available to the average surgeon. As an inventor, he designed many surgical instruments, including a circular retractor holder, as well as a universal golf club, a circular tennis racquet and a modified bicycle seat. CONCLUSIONS: Sir Denis Browne was an innovator and nonconformist with regard to the traditional teaching and practice of pediatric surgery and urology of his day. His innovations in surgical theory, technique and instrumentation resulted in major contributions to the advancement of urology as well as pediatric surgery. PMID- 20005180 TI - The effects of maternal labour analgesia on the fetus. AB - Maternal labour pain and stress are associated with progressive fetal metabolic acidosis. Systemic opioid analgesia does little to mitigate this stress, but opioids readily cross the placenta and cause fetal-neonatal depression and impair breast feeding. Pethidine remains the most widely used, but alternatives, with the possible exception of remifentanil, have little more to offer. Inhalational analgesia using Entonox is more effective and, being rapidly exhaled by the newborn, is less likely to produce lasting depression. Neuraxial analgesia has maternal physiological and biochemical effects, some of which are potentially detrimental and some favourable to the fetus. Actual neonatal outcome, however, suggests that benefits outweigh detrimental influences. Meta-analysis demonstrates that Apgar score is better after epidural than systemic opioid analgesia, while neonatal acid-base balance is improved by epidural compared to systemic analgesia and even compared to no analgesia. Successful breast feeding is dependent on many factors, therefore randomized trials are required to elucidate the effect of labour analgesia. PMID- 20005181 TI - Point mutations are causing progeroid phenotypes in the mtDNA mutator mouse. PMID- 20005182 TI - Non-specific DNA binding interferes with the efficient excision of oxidative lesions from chromatin by the human DNA glycosylase, NEIL1. AB - Although DNA in eukaryotes is packaged in nucleosomes, it remains vulnerable to oxidative damage that can result from normal cellular metabolism, ionizing radiation, and various chemical agents. Oxidatively damaged DNA is repaired in a stepwise fashion via the base excision repair (BER) pathway, which begins with the excision of damaged bases by DNA glycosylases. We reported recently that the human DNA glycosylase hNTH1 (human Endonuclease III), a member of the HhH GpG superfamily of glycosylases, can excise thymine glycol lesions from nucleosomes without requiring or inducing nucleosome disruption; optimally oriented lesions are excised with an efficiency approaching that seen for naked DNA [1]. To determine if this property is shared by human DNA glycoylases in the Fpg/Nei family, we investigated the activity of NEIL1 on defined nucleosome substrates. We report here that the cellular concentrations and apparent k(cat)/K(M) ratios for hNTH1 and NEIL1 are similar. Additionally, after adjustment for non-specific DNA binding, hNTH1 and NEIL1 proved to have similar intrinsic activities toward nucleosome substrates. However, NEIL1 and hNTH1 differ in that NEIL1 binds undamaged DNA far more avidly than hNTH1. As a result, hNTH1 is able to excise both accessible and sterically occluded lesions from nucleosomes at physiological concentrations, while the high non-specific DNA affinity of NEIL1 would likely hinder its ability to process sterically occluded lesions in cells. These results suggest that, in vivo, NEIL1 functions either at nucleosome-free regions (such as those near replication forks) or with cofactors that limit its non-specific binding to DNA. PMID- 20005184 TI - Simultaneous determination of ABT-888, a poly (ADP-ribose) polymerase inhibitor, and its metabolite in human plasma by liquid chromatography/tandem mass spectrometry. AB - A reversed-phase liquid chromatography coupled to tandem mass spectrometry (LC MS/MS) method was developed and validated for simultaneous determination of ABT 888 and its major metabolite (M8) in human plasma. Sample preparation involved a liquid-liquid extraction by the addition of 0.25 ml of plasma with 10 microl of 1 M NaOH and 1.0 ml ethyl acetate containing 50 ng/ml of the internal standard zileuton. The analytes were separated on a Waters XBridge C(18) column using a gradient mobile phase consisting of methanol/water containing 0.45% formic acid at the flow rate of 0.2 ml/min. The analytes were monitored by tandem mass spectrometry with electrospray positive ionization. Linear calibration curves were generated over the ABT-888 and M8 concentration ranges of 1-2000 ng/ml in human plasma. The lower limits of quantitation (LLOQ) were 1 ng/ml for both ABT 888 and M8 in human plasma. The accuracy and within- and between-day precisions were within the generally accepted criteria for bioanalytical method (<15%). This method was successfully employed to characterize the plasma concentration-time profile of ABT-888 after its oral administration in cancer patients. PMID- 20005183 TI - Use of Drosophila deoxynucleoside kinase to study mechanism of toxicity and mutagenicity of deoxycytidine analogs in Escherichia coli. AB - Most bacteria, including Escherichia coli, lack an enzyme that can phosphorylate deoxycytidine and its analogs. Consequently, most studies of toxicity and mutagenicity of cytosine analogs use ribonucleosides such as 5-azacytidine (AzaC) and zebularine (Zeb) instead of their deoxynucleoside forms, 5-aza-2' deoxycytidine (AzadC) and 2'-deoxy-zebularine (dZeb). The former analogs are incorporated into both RNA and DNA creating complex physiological responses in cells. To circumvent this problem, we introduced into E. coli the Drosophila deoxynucleoside kinase (Dm-dNK), which has a relaxed substrate specificity, and tested these cells for sensitivity to AzadC and dZeb. We find that Dm-dNK expression increases substantially sensitivity of cells to these analogs and dZeb is very mutagenic in cells expressing the kinase. Furthermore, toxicity of dZeb in these cells requires DNA mismatch correction system suggesting a mechanism for its toxicity and mutagenicity. The fluorescence properties of dZeb were used to quantify the amount of this analog incorporated into cellular DNA of mismatch repair-deficient cells expressing Dm-dNK and the results showed that in a mismatch correction-defective strain a high percentage of DNA bases may be replaced with the analog without long term toxic effects. This study demonstrates that the mechanism by which Zeb and dZeb cause cell death is fundamentally different than the mechanism of toxicity of AzaC and AzadC. It also opens up a new way to study the mechanism of action of deoxycytidine analogs that are used in anticancer chemotherapy. PMID- 20005185 TI - Simultaneous HPLC-UV analysis of rufinamide, zonisamide, lamotrigine, oxcarbazepine monohydroxy derivative and felbamate in deproteinized plasma of patients with epilepsy. AB - We present an implementation of a method we previously reported allowing the newer antiepileptic drugs (AEDs) rufinamide (RFN) and zonisamide (ZNS) to be simultaneously determined with lamotrigine (LTG), oxcarbazepine's (OXC) main active metabolite monohydroxycarbamazepine (MHD) and felbamate (FBM) in plasma of patients with epilepsy using high performance liquid chromatography (HPLC) with UV detection. Plasma samples (250 microL) were deproteinized by 1 mL acetonitrile spiked with citalopram as internal standard (I.S.). HPLC analysis was carried out on a Synergi 4 microm Hydro-RP, 250 mm x 4.6 mm I.D. column. The mobile phase was a mixture of potassium dihydrogen phosphate buffer (50 mM, pH 4.5), acetonitrile and methanol (65:26.2:8.8, v/v/v) at an isocratic flow rate of 0.8 mL/min. The UV detector was set at 210 nm. The chromatographic run lasted 19 min. Commonly coprescribed AEDs did not interfere with the assay. Calibration curves were linear for both AEDs over a range of 2-40 microg/mL for RFN and 2-80 microg/mL for ZNS. The limit of quantitation was 2 microg/mL for both analytes and the absolute recovery ranged from 97% to 103% for RFN, ZNS and the I.S. Intra- and interassay precision and accuracy were lower than 10% at all tested concentrations. The present study describes the first simple and validated method for RFN determination in plasma of patients with epilepsy. By grouping different new AEDs in the same assay the method can be advantageous for therapeutic drug monitoring (TDM). PMID- 20005187 TI - Comment to "The efficacy of Helicobacter pylori eradication regimen with and without vitamin C supplementation". PMID- 20005186 TI - Up-regulated proteins in the fluid bathing the tumour cell microenvironment as potential serological markers for early detection of cancer of the breast. AB - Breast cancer is by far the most common diagnosed form of cancer and the leading cause of cancer death in women today. Clinically useful biomarkers for early detection of breast cancer could lead to a significant reduction in mortality. Here we describe a detailed analysis using gel-based proteomics in combination with mass spectrometry and immunohistochemistry (IHC) of the tumour interstitial fluids (TIF) and normal interstitial fluids (NIF) collected from 69 prospective breast cancer patients. The goal of this study was to identify abundant cancer up regulated proteins that are externalised by cells in the tumour microenvironment of most if not all these lesions. To this end, we applied a phased biomarker discovery research strategy to the analysis of these samples rather than comparing all samples among each other, with inherent inter and intra-sample variability problems. To this end, we chose to use samples derived from a single tumour/benign tissue pair (patient 46, triple negative tumour), for which we had well-matched samples in terms of epithelial cell numbers, to generate the initial dataset. In this first phase we found 110 proteins that were up-regulated by a factor of 2 or more in the TIF, some of which were confirmed by IHC. In the second phase, we carried out a systematic computer assisted analysis of the 2D gels of the remaining 68 TIF samples in order to identify TIF 46 up-regulated proteins that were deregulated in 90% or more of all the available TIFs, thus representing common breast cancer markers. This second phase singled out a set of 26 breast cancer markers, most of which were also identified by a complementary analysis using LC-MS/MS. The expression of calreticulin, cellular retinoic acid binding protein II, chloride intracellular channel protein 1, EF-1-beta, galectin 1, peroxiredoxin-2, platelet-derived endothelial cell growth factor, protein disulfide isomerase and ubiquitin carboxyl-terminal hydrolase 5 were further validated using a tissue microarray containing 70 malignant breast carcinomas of various grades of atypia. A significant number of these proteins have already been detected in the blood/plasma/secretome by others. The next steps, which include biomarker prioritization based on the hierarchal evaluation of these markers, antibody and antigen development, assay development, analytical validation, and preliminary testing in the blood of healthy and breast cancer patients, are discussed. PMID- 20005188 TI - A new practical alternative for tumoural gastrointestinal bleeding: Ankaferd blood stopper. PMID- 20005190 TI - [Myotonic dystrophy. 18 years experience in a neuropaediatric clinic]. AB - INTRODUCTION: Myotonic dystrophy is a highly variable autosomic dominant inherited multisystemic disease. We review our 18 years experience with patients suffering from this disease. RESULTS: Eleven patients were identified following a molecular genetic study: 2 patients died, 5 are still under control, 2 are being controlled in another Centre, and 3 dropped out. Three of them were relatives. Seven newborns started with hypotonic symptoms in the neonatal period, with hypotonic symptoms, of which 4 had foetal suffering. One child was diagnosed at age of 3 due to her father being affected. One girl was seen at age of 10 due to stiffness and tightening of her hands for years. One boy, aged 5, was examined due to abnormal hands posture, and a 4 year old child due to psychomotor delay. Associated disorders: 7 children with psychomotor delay, 2 cases of cataracts, 1 case of diabetes type I, 3 cases of hypercholesterolemia, 1 abdominal sarcoma, 1 case of femur and hip fracture, 2 cases of interatrial communication. The diagnostic was made in 5 cases by a clinic due to mother-son relation phenotype, in 3 cases after the family diagnosis and in another 3 cases non-congenital symptoms exclusively in the child's clinic. DISCUSSION: In our experience, myotonic dystrophy is uncommon; it is often congenital, and is associated with perinatal suffering. Genetics can identify or exclude the process. This must be done on newborns who are hypotonic for an unknown reason. It should be suspected in a child who presents with motor abnormalities in the fingers and hands. PMID- 20005191 TI - [Spontaneous resolution of a pseudoaneurysm following arterial puncture]. PMID- 20005189 TI - Gastro-protective strategies in primary care in Italy: the "Gas.Pro." survey. AB - INTRODUCTION: Risk of gastrointestinal injury is relevant among users of anti inflammatory or cardio-protective drugs. Adequate gastro-protection is warranted in high-risk patients. AIM: To assess the perceptions and practices of Italian primary care physicians regarding gastro-protective strategies. METHODS: Nationwide cross-sectional observational study. A 14-question survey questionnaire was administered to 112 primary care physicians throughout Italy. Data collection covered consecutive outpatient candidates for the prescription of a potentially GI harmful medication, observed in the physicians' office over a 3 week period. RESULTS: Cohort included 3943 cases (2489 naive and 1463 chronic NSAID/ASA users). Mean age and prevalence of cardiovascular comorbidity were significantly higher in the latter subgroup. Non-selective NSAIDs and low-dose aspirin were the most commonly prescribed drugs. Combined NSAIDS/ASA plus steroids/anticoagulant/antiplatelets were recorded in 161 cases. Helicobacter pylori status was known in only 38% of naive and 33.2% of chronic users, being negative in 85.3% and 89.5%, respectively. When positive, H. pylori was eradicated by almost all physicians (97.9%), but in case of unknown H. pylori status, the presence of infection was investigated in only 8.6% and 14.9% of patients in the two subgroups. Gastro-protection was endorsed in 80.7% of patients, mostly PPIs (91%). In patients aged over 70, pantoprazole and lansoprazole were the preferred gastro-protective agents. CONCLUSIONS: There is a significant over-use of gastro-protection in the primary care setting in Italy and the role H. pylori is largely overlooked. Educational efforts should be directed to a more targeted gastro-protection only for at-risk patients as well as improved adherence to recommendations for testing and treating H. pylori infection. PMID- 20005192 TI - Obesity in young women during pregnancy. PMID- 20005193 TI - Biodegradable fibrin conduit promotes long-term regeneration after peripheral nerve injury in adult rats. AB - Peripheral nerve injuries are often associated with loss of nerve tissue and require autologous nerve grafts to provide a physical substrate for axonal growth. Biosynthetic neural conduits could be an alternative treatment strategy in such injuries. The present study investigates the long-term effects of a tubular fibrin conduit on neuronal regeneration, axonal sprouting and recovery of muscle weight following peripheral nerve injury and repair in adult rats. Sciatic axotomy was performed proximally in the thigh to create a 10-mm gap between the nerve stumps. The injury gap was bridged by using a 14-mm-long fibrin glue conduit, entubulating 2 mm of the nerve stump at each end. A reversed autologous nerve graft was used as a control. The regenerative response from sensory and motor neurones was evaluated following retrograde labelling with Fast Blue fluorescent tracer. In control experiments, at 16 weeks following peripheral nerve grafting, 5184 (+/-574 standard error of mean (SEM)) sensory dorsal root ganglion neurones and 1001 (+/-37 SEM) spinal motor neurones regenerated across the distal nerve-graft interface. The fibrin conduit promoted regeneration of 60% of sensory neurones and 52% of motor neurones when compared to the control group. The total number of myelinated axons in the distal nerve stump in the fibrin conduit group reached 86% of the control and the weight of gastrocnemius and soleus muscles recovered to 82% and 89% of the controls, respectively. The present results suggest that a tubular fibrin conduit can be used to promote neuronal regeneration following peripheral nerve injury. PMID- 20005194 TI - Complete breast reconstruction with autologous fat graft - a case report. AB - Large-volume fat grafting can be used as an alternative method of breast reconstruction in selected cases. A pleasing aesthetic result as well as its longevity is possible. PMID- 20005195 TI - Routine drainage in reduction mammaplasty: an evidence-based analysis. PMID- 20005196 TI - Evaluation of the performance of CareStart Malaria Pf/Pv Combo rapid diagnostic test for the diagnosis of malaria in Jimma, southwestern Ethiopia. AB - OBJECTIVE: To evaluate the diagnostic performance of CareStart Malaria Pf/Pv Combo test relative to microscopy, for the diagnosis of falciparum and vivax malaria in Ethiopia. METHODS: Two hundred and forty febrile patients visiting the Serbo health center in Jimma zone, southwestern Ethiopia, were involved in this study in 2008. Giemsa-stained thin and thick blood smears were prepared and microscopically examined under a 100x oil immersion microscope objective for Plasmodium species identification and determination of parasitemia respectively. CareStart Malaria Pf/Pv Combo test was performed as per the manufacturers' instruction. FINDINGS: The validity of CareStart Malaria Pf/Pv Combo test for the diagnosis of Plasmodium was very good with a sensitivity of 95.8%, specificity of 100%, positive predictive value of 100% and negative predictive value of 96%. The test performed equally well for the identification of Plasmodium falciparum and P. vivax. The diagnostic performance of this CareStart test is comparable to light microscopy of thin and thick blood smears. CONCLUSION: Although CareStart Malaria Pf/Pv Combo test and blood microscopy have comparable diagnostic performance for Plasmodium detection, the CareStart test has the added advantage of being simple to interpret, cost-efficient, and hence it is preferable to use this rapid diagnostic test for malaria diagnosis in areas where microscopy is not accessible and during times of malaria epidemics that are observed approximately every 4-5 years in Ethiopia. PMID- 20005197 TI - Structural insights into the substrate recognition properties of beta glucosidase. AB - Beta-glucosidase enzymes (EC 3.2.1-3.2.3) hydrolyze sugars and are implicated in a wide spectrum of biological processes. Recently, we reported that beta glucosidase has varied kinetic parameters for the natural and synthetic substrates [K.H Nam, S.J. Kim, M.Y. Kim, J.H. Kim, T.S. Yeo, C.M. Lee, H.K Jun, K.Y. Hwang. Crystal structure of engineered beta-glucosidase from a soil metagenome, Proteins 73 (2008) 788-793]. However, an understanding of the kinetic values of beta-glucosidase has not yet enabled the elucidation of its molecular function. Here, we report the X-ray crystal structure of beta-glucosidase with a glucose and cellobiose fragment from uncultured soil metagenome. From the various crystals, we obtained the pre-reaction (native), intermediate (disaccharide cleavage) and post-reaction (glucose binding) states of the active site pocket. These structures provide snapshots of the catalytic processing of beta glucosidase. In addition, the intermediate state of the crystal structure provides insight into the substrate specificity of beta-glucosidase. These structural studies will facilitate elucidation of the architectural mechanism responsible for the substrate recognition of beta-glucosidase. PMID- 20005198 TI - Amphipathic peptides can act as an anticoagulant by competing with phospholipid membranes for blood coagulation factors. AB - An ideal amphipathic peptide (IAP), composed of simply lysine and leucine residues in a 1:2 ratio (K(7)L(15)), specifically prolongs in vitro coagulation assays that use phospholipids, such as the activated partial thromboplastin time (APTT). The main hypothesis of the present work is that IAP's anticoagulant effect occurs by competing with phospholipid membranes in in vitro coagulation reactions. We verified this hypothesis by employing different phospholipid dependent coagulation assays, such as the APTT, the dilute prothrombin time (dPT) and the dilute Russell viper venom time (dRVVT) with both low and high amounts of phospholipids. We show that coagulation times are prolonged by IAP in a concentration-dependent manner, and that this prolongation is abrogated by adding excess phospholipid, demonstrating a phospholipid dependence for this inhibition. Using an ELISA-based binding assay, we show IAP inhibits the binding of one of the vitamin K-dependent coagulation factors, factor X, to phospholipid membranes. This is further confirmed with fluorescence spectroscopy, where the interaction of IAP and factor X is inhibited by phospholipid. In summary, this work demonstrates that IAP can act as an anticoagulant by impairing the interaction of coagulation factors with phospholipid membranes and provides a paradigm for the development of novel anticoagulants. PMID- 20005199 TI - Identification and characterization of small sub-genomic RNAs in dengue 1-4 virus infected cell cultures and tissues. AB - Dengue virus (DV) contains a single-stranded, positive-sense RNA genome, and the 3' non-coding regions (NCRs) have been demonstrated to play crucial roles in viral RNA replication and translation initiation. In this study, we report the presence of a species of small sub-genomic RNA (sgRNA) derived from the 3' NCR in DV-infected cell cultures and animal tissues. Further experiments demonstrate that these sgRNAs range from 390 to 430 nt in length and that they accumulate together with viral genomic RNA during replication. Sequence analysis and secondary structure prediction indicate that the stem-loop structures within the 3' NCR are responsible for the production of DV sgRNAs. Unexpectedly, a unique DV isolate produced three species of sgRNAs, compared to only one in the other isolates. Collectively, the structural and biochemical properties of DV sgRNAs warrant further study of their biological functions during the viral life cycle and pathogenesis. PMID- 20005200 TI - Molecular characterization and expression of the low-density lipoprotein receptor related protein-10, a new member of the LDLR gene family. AB - We report the characterization of a new member of the low-density lipoprotein receptor (LDLR) gene family designated LRP10. Human LRP10 cDNA encodes a 1905 amino acid type I membrane protein consisting of five functional domains characteristic of the LDLR gene family. CHO-ldlA7 cells transfected with human LRP10 cDNA bound LDLR-associated protein, but not beta-VLDL and HDL. Human LRP10 transcripts were primarily found in the brain, muscle and heart. In situ hybridization of the rat brain showed that the transcripts were intensely present in the cerebral cortex, hippocampus, choroid plexus, ependyma and granular layer. In the developing rat brain, transcript levels gradually increased from postnatal day 1 to 20. Immunofluorescence analysis indicated that LRP10 was observed in the ventricular zone of the embryonic day 14.5 mouse cerebral cortex. The present studies suggest that LRP10 may play a significant role in the brain physiology other than lipoprotein metabolism. PMID- 20005201 TI - Nitric oxide and thioredoxin type 1 modulate the activity of caspase 8 in HepG2 cells. AB - Herein, we report that nitric oxide (NO) and the thioredoxin/thioredoxin reductase system affect the activity of caspase 8 in HepG2 cells. Exposure of cells to NO resulted in inhibition of caspase 8, while a subsequent incubation of the cells in NO-free medium resulted in spontaneous reactivation of the protease. The latter process was inhibited in thioredoxin reductase-deficient HepG2 cells, in which, however, lipoic acid markedly reactivated caspase 8. The data obtained suggest that extrinsic apoptosis can be subjected to redox regulation before induction of proteolytic damage by caspase 3. PMID- 20005202 TI - Fibulin-3 negatively regulates chondrocyte differentiation. AB - Fibulin-3 is a member of the fibulin family that has been newly recognized as extracellular matrix proteins. We assessed the effects of fibulin-3 overexpression on chondrocyte differentiation using the clonal murine cell line ATDC5. The ATDC5-FBLN3 stably expressing fibulin-3 protein was spindle-shaped cell compared to the ATDC5-mock with plump cell. The cell growth in the ATDC5 FBLN3 was accelerated in comparison to that in the ATDC5-mock. The ATDC5-FBLN3 was not stained by Alcian blue, nor was there any cartilage aggregate formed after the induction of chondrogenic differentiation. The expression of type II collagen, aggrecan, and type X collagen was completely suppressed in ATDC5-FBLN3 even after the induction of differentiation. The overexpression of fibulin-3 reduced the expression of Sox5 and Sox6, while it maintained the expression of Sox9. These findings suggest that fibulin-3 may play an important role as a negative regulator of chondrocyte differentiation. PMID- 20005203 TI - Biophysical characterization reveals structural disorder in the developmental transcriptional regulator LBH. AB - Limb-bud and heart (LBH) is a key transcriptional regulator in vertebrates with pivotal roles in embryonic development and human disease. Herein, using a diverse array of biophysical techniques, we report the first structural characterization of LBH pertinent to its biological function. Our data reveal that LBH is structurally disordered with no discernable secondary or tertiary structure and exudes rod-like properties in solution. Consistent with these observations, we also demonstrate that LBH is conformationally flexible and thus may be capable of adapting distinct conformations under specific physiological contexts. We propose that LBH is a member of the intrinsically disordered protein (IDP) family, and that conformational plasticity may play a significant role in modulating LBH dependent transcriptional processes. PMID- 20005204 TI - Proteomic analysis of astrocytic secretion that regulates neurogenesis using quantitative amine-specific isobaric tagging. AB - Astrocytes are essential components of neurogenic niches that affect neurogenesis through membrane association and/or the release of soluble factors. To identify factors released from astrocytes that could regulate neural stem cell differentiation and proliferation, we used mild oxygen-glucose deprivation (OGD) to inhibit the secretory capacity of astrocytes. Using the Transwell co-culture system, we found that OGD-treated astrocytes could not promote neural stem cell differentiation and proliferation. Next, isobaric tagging for the relative and absolute quantitation (iTRAQ) proteomics techniques was performed to identify the proteins in the supernatants of astrocytes (with or without OGD). Through a multi step analysis and gene ontology classification, 130 extracellular proteins were identified, most of which were involved in neuronal development, the inflammatory response, extracellular matrix composition and supportive functions. Of these proteins, 44 had never been reported to be produced by astrocytes. Using ProteinPilot software analysis, we found that 60 extracellular proteins were significantly altered (27 upregulated and 33 downregulated) in the supernatant of OGD-treated astrocytes. Among these proteins, 7 have been reported to be able to regulate neurogenesis, while others may have the potential to regulate neurogenesis. This study profiles the major proteins released by astrocytes, which play important roles in the modulation of neurogenesis. PMID- 20005205 TI - An evolved Escherichia coli strain for producing hydrogen and ethanol from glycerol. AB - Glycerol is an attractive feedstock for biofuels since it accumulates as a byproduct during biodiesel operations; hence, here we consider converting glycerol to hydrogen using the formate hydrogen lyase system of Escherichia coli which converts pyruvate to hydrogen. Starting with E. coli BW25113 frdC that lacks fumarate reductase (this mutation reduces repression of hydrogen synthesis during glycerol fermentation) and by using both adaptive evolution and chemical mutagenesis combined with a selection method based on increased growth on glycerol, we obtained an improved strain, HW2, that produces 20-fold more hydrogen in glycerol medium (0.68+/-0.16 mmol/L/h). HW2 also grows 5-fold faster (0.25+/-0.01/h) than BW25113 frdC on glycerol, so it achieves a reasonable anaerobic growth rate. Corroborating the increase in hydrogen production, glycerol dehydrogenase activity in HW2 increased 4-fold compared to BW25113 frdC. In addition, a whole-transcriptome study revealed that several pathways that would decrease hydrogen yields were repressed in HW2 (fbp, focA, and gatYZ) while a beneficial pathway which encodes enolase was induced. Ethanol production was also increased 5-fold in the evolved HW2 strain. PMID- 20005206 TI - Essential role of TRPC1 channels in cardiomyoblasts hypertrophy mediated by 5 HT2A serotonin receptors. AB - Serotonin (5-HT) participates in the development of cardiac hypertrophy through 5 HT(2A) serotonin receptors. The hypertrophic growth of cardiomyoblasts induced by 5-HT(2A) receptors involves the activation of the Ca(2+) responsive calcineurin/NFAT pathway. However, the mechanism whereby NFAT is activated by 5 HT(2A) receptors remains indeterminate. In this study, we examined whether transient receptor potential canonical (TRPC) channels participate in NFAT activation and hypertrophic response triggered by 5-HT. We demonstrate that TRPC1 expression is upregulated in 5-HT-treated rat cardiomyoblasts whereas TRPC6 is induced in a mouse model of heart hypertrophy. Moreover, TRPC1 knockdown by small interfering RNA inhibits NFAT activation and hypertrophic response mediated by 5 HT(2A) receptors. These findings provide new insights about a mechanistic basis for the activation of the calcineurin/NFAT pathway by 5-HT(2A) receptors and highlight the critical role of TRPC1 in the development of cardiac hypertrophy. PMID- 20005207 TI - Overexpressed proteins may act as mops removing their ligands from the host cells: a case study of calf PNP. AB - Calf purine nucleoside phosphorylase (PNP) was overexpressed in Escherichia coli. The basic kinetic parameters of recombinant PNP were found to be similar to the values published previously for non-recombinant PNP from calf spleen. However, upon titration of the recombinant enzyme with the tight-binding multisubstrate analogue inhibitor DFPP-DG, endothermic as well as exothermic signals were obtained. This was not the case for PNP isolated from calf spleen for which only the endothermic process was observed. Further calorimetric titrations of the recombinant and non-recombinant enzyme with its potent and moderate ligands, and studied involving partial inactivation of the enzyme, lead to the conclusion that a part of the recombinant enzyme forms a complex with its product, hypoxanthine, although hypoxanthine was not present at any purification stage except for its natural occurrence in E. coli cells. Binding of hypoxanthine is accompanied with a large negative change of the free enthalpy, and therefore the replacement of this compound by DFPP-DG yields positive heat signal. Our data obtained with calf PNP indicate that similar processes--moping of ligands from the host cells--may take place in the case of other proteins with high overexpression yield. PMID- 20005208 TI - Carbon monoxide enhance colonic epithelial restitution via FGF15 derived from colonic myofibroblasts. AB - Carbon monoxide (CO) has been reported to ameliorate colonic inflammation and improve experimental colitis. It is well known that mucosal restitution is important to improve colitis as well as reduction of mucosal inflammation. However, it has not been clear whether CO effects to colonic mucosal restitution or not. In general, colonic myofibroblast (MF) has been reported to play an important role of colonic epithelial cell restitution via constitutive secretion of TGF-beta. In this study, we showed CO (supplied by CO-releasing molecule; CORM) treated MF conditioned medium enhanced colonic epithelial cell (YAMC) restitution and we determined gene expression in colonic MF treated with CO using microRNA. The microRNA array suggested that miR-710 was significantly reduced in MF by CO treatment and the target gene of miR-710 is determined to fibroblast growth factor (FGF)15. The CO treated MF conditioned medium which FGF15 expression was silenced extinguished the enhancement effect of epithelial cell restitution. Our findings demonstrate that CO treatment to MF increased FGF15 expression via inhibition of miR-710 and FGF15 enhanced colonic epithelial cell restitution. PMID- 20005209 TI - The UV-filter benzophenone-1 inhibits 17beta-hydroxysteroid dehydrogenase type 3: Virtual screening as a strategy to identify potential endocrine disrupting chemicals. AB - The prevalence of male reproductive disorders and testicular cancer is steadily increasing. Because the exposure to chemicals disrupting natural hormone action has been associated with these diseases, it is important to identify endocrine disrupting chemicals (EDCs) and their targets of action. Here, a 3D-structural database that can be applied for virtual screening approaches to facilitate the identification of EDCs was constructed. The database was screened using pharmacophores of 17beta-hydroxysteroid dehydrogenase type 3 (17beta-HSD3), which catalyzes the last step of testosterone synthesis in testicular Leydig cells and plays an essential role during male sexual development. Among other chemicals, benzophenone (BP) UV-filters were predicted as potential 17beta-HSD3 inhibitors. Biological analyses revealed (2,4-dihydroxyphenyl)-phenylmethanone (also known as benzophenone-1, BP-1) as an inhibitor of human 17beta-HSD3 (IC(50) 1.05microM). BP-1 also efficiently blocked conversion of androstenedione to testosterone by mouse and rat 17beta-HSD3 in whole-organ enzyme assays. Moreover, BP-1 antagonized the testosterone-dependent activation of androgen receptors (IC(50) 5.7microM), suggesting synergistic anti-androgenic effects of BP-1 by preventing testosterone formation and blocking receptor activation. In addition, analyses of several commonly used UV-filters on estrogen- and androgen-metabolizing 17beta HSD enzymes revealed 3-benzylidene camphor (3-BC) and 4-methylbenzylidene camphor (4-MBC) as low micromolar 17beta-HSD2 inhibitors. In conclusion, screening of virtual chemical structure libraries can facilitate the identification of compounds interfering with hormone action. The potential disruption of 17beta-HSD enzyme function by the UV-filters BP-1, 3-BC and 4-MBC requires further investigation and should be considered for safety assessment of these chemicals. PMID- 20005210 TI - Emodin enhances sensitivity of gallbladder cancer cells to platinum drugs via glutathion depletion and MRP1 downregulation. AB - Glutathione conjugation and transportation of glutathione conjugates of anticancer drugs out of cells are important for detoxification of many anticancer drugs. Inhibition of this detoxification system has recently been proposed as a strategy to treat drug-resistant solid tumors. Gallbladder carcinoma is resistant to many anticancer drugs, therefore, it is needed to develop a novel strategy for cancer therapy. In the present study, we tested the effect of emodin (1,3,8 trihydroxy-6-methylanthraquinone), a reactive oxygen species (ROS) generator reported by our group previously, in combination with cisplatin (CDDP), carboplatin (CBP) or oxaliplatin in treating the gallbladder carcinoma cell line SGC996. Our results showed that co-treatment with emodin could remarkably enhance chemosensitivity of SGC996 cells in comparison with cisplatin, carboplatin or oxaliplatin treatment alone. We found that the mechanisms may be attributed to reduction of glutathione level, and downregulation of multidrug resistance related protein 1 (MRP1) expression in SGC996 cells. The experiments on tumor bearing mice showed that emodin/cisplatin co-treatment inhibited the tumor growth in vivo via increasing tumor cell apoptosis and downregulating MRP1 expression. In conclusion, emodin can work as an adjunct to enhance the anticancer effect of platinum drugs in gallbladder cancer cells via ROS-related mechanisms. PMID- 20005211 TI - Reductive activation of the prodrug 1,2-bis(methylsulfonyl)-1-(2-chloroethyl)-2 [[1-(4-nitrophenyl)ethoxy]carbonyl]hydrazine (KS119) selectively occurs in oxygen deficient cells and overcomes O(6)-alkylguanine-DNA alkyltransferase mediated KS119 tumor cell resistance. AB - 1,2-Bis(methylsulfonyl)-1-(2-chloroethyl)-2-[[1-(4 nitrophenyl)ethoxy]carbonyl]hydrazine (KS119) is a prodrug of the 1,2 bis(sulfonyl)hydrazine class of antineoplastic agents designed to exploit the oxygen-deficient regions of cancerous tissue. Thus, under reductive conditions in hypoxic cells this agent decomposes to produce the reactive intermediate 1,2 bis(methylsulfonyl)-1-(2-chloroethyl)hydrazine (90CE), which in turn generates products that alkylate the O(6)-position of guanine in DNA. Comparison of the cytotoxicity of KS119 in cultured cells lacking O(6)-alkylguanine-DNA alkyltransferase (AGT) to an agent such as Onrigin, which through base catalyzed activation produces the same critical DNA G-C cross-link lesions by the generation of 90CE, indicates that KS119 is substantially more potent than Onrigin under conditions of oxygen deficiency, despite being incompletely activated. In cell lines expressing relatively large amounts of AGT, the design of the prodrug KS119, which requires intracellular activation by reductase enzymes to produce a cytotoxic effect, results in an ability to overcome resistance derived from the expression of AGT. This appears to derive from the ability of a small portion of the chloroethylating species produced by the activation of KS119 to slip through the cellular protection afforded by AGT to generate the few DNA G-C cross-links that are required for tumor cell lethality. The findings also demonstrate that activation of KS119 under oxygen-deficient conditions is ubiquitous, occurring in all of the cell lines tested thus far, suggesting that the enzymes required for reductive activation of this agent are widely distributed in many different tumor types. PMID- 20005212 TI - Identification of small molecule inhibitors of pyruvate kinase M2. AB - A common feature of tumors arising from diverse tissue types is a reliance on aerobic glycolysis for glucose metabolism. This metabolic difference between cancer cells and normal cells could be exploited for therapeutic benefit in patients. Cancer cells universally express the M2 isoform of the glycolytic enzyme pyruvate kinase (PKM2), and previous work has demonstrated that PKM2 expression is necessary for aerobic glycolysis and cell proliferation in vivo. Because most normal tissues express an isoform of pyruvate kinase other than PKM2, selective targeting of PKM2 provides an opportunity to target cell metabolism for cancer therapy. PKM2 has an identical catalytic site as the related M1 splice variant (PKM1). However, isoform selective inhibition is possible as PKM2 contains a unique region for allosteric regulation. We have screened a library of greater than 1,00,000 small molecules to identify such inhibitors. The inhibitors identified for PKM2 fell primarily into three distinct structural classes. The most potent PKM2 inhibitor resulted in decreased glycolysis and increased cell death following loss of growth factor signaling. At least part of this effect was due to on-target PKM2 inhibition as less cell death was observed in cells engineered to express PKM1. These data suggest that isoform selective inhibition of PKM2 with small molecules is feasible and support the hypothesis that inhibition of glucose metabolism in cancer cells is a viable strategy to treat human malignancy. PMID- 20005213 TI - Inhibition of epidermal growth factor receptor over-expressing cancer cells by the aphorphine-type isoquinoline alkaloid, dicentrine. AB - The extraordinary relevance of EGFR in tumour biology makes it an exquisite molecular target for tumour therapy. Despite considerable success with these EGFR tyrosine kinase inhibitors in cancer therapy, resistance against these chemical compounds develops owing to the selection of point-mutated variants of EGFR. Therefore, there is an urgent need for the identification of novel EGFR tyrosine kinase inhibitors for treating tumours with such EGFR mutants. We found a preferential cytotoxicity of dicentrine towards U87MG.DeltaEGFR-transduced with a constitutively deletion-activated EGFR expression vector as compared to non transduced wild-type U87MG cells. As determined by microarray-based mRNA expression profiling, this preferential cytotoxicity was accompanied with an activation of BRCA1-mediated DNA damage response, p53 signalling, G1/S and G2/M cell cycle regulation, and aryl hydrocarbon receptor pathways. The activation of these signalling routes might be explained by the fact that dicentrine intercalates DNA and induces DNA strand break by inhibition of DNA topoisomerases. The cell cycle might be arrested by dicentrine-induced DNA lesions. PMID- 20005215 TI - Behavioral, neuroimaging, and computational evidence for perceptual caching in repetition priming. AB - Repetition priming (RP) is a form of learning, whereby classification or identification performance is improved with item repetition. Various theories have been proposed to understand the basis of RP, including alterations in the representation of an object and associative stimulus-response bindings. There remain several aspects of RP that are still poorly understood, and it is unclear whether previous theories only apply to well-established object representations. This paper integrates behavioral, neuroimaging, and computational modeling experiments in a new RP study using novel objects. Behavioral and neuroimaging results were inconsistent with existing theories of RP, thus a new perceptual memory-based caching mechanism is formalized using computational modeling. The model instantiates a viable neural mechanism that not only accounts for the pattern seen in this experiment but also provides a plausible explanation for previous results that demonstrated residual priming after associative linkages were disrupted. Altogether, the current work helps advance our understanding of how brain utilizes repetition for faster information processing. PMID- 20005214 TI - Feature selection in the human brain: electrophysiological correlates of sensory enhancement and feature integration. AB - This study examined the latency and amplitude of cortical processes associated with feature-based visual selective attention, using frequency-domain and time domain measures derived from dense-array electroencephalography. Participants were asked to identify targets based on conjunctions of three types of object features (color, size, and completeness). This procedure aimed to examine (1) the modulation of sensory responses to one or more stimulus features characterizing an object and (2) the facilitation and reduction effects associated with competing features, attended and unattended, in the same object. The selection negativity, an event-related potential measure of sensory amplification for attended features, showed a parametric increase of amplitude as a function of the number of attended features. Late oscillations in the gamma band range were also smaller for stimuli with one or more non-attended visual features but were enhanced for stimuli sharing the overall gestalt with the target. The latency of this late gamma modulation was delayed when two target features were combined, compared to one single discriminative feature. Latency analyses also showed that late bursts of induced high-frequency oscillatory activity peaked around 60 ms later than the selection negativity. Oscillatory activity reflected both selective amplification and competition between object features. These results suggest that sensory amplification of selected features is followed by integrative processing in more widespread networks. Oscillatory activity in these networks is reduced by distraction and is enhanced when attended features can be mapped to specific action. PMID- 20005216 TI - Branching patterns for arterioles and venules of the human cerebral cortex. AB - Branching patterns of microvascular networks influence vascular resistance and allow control of peripheral flow distribution. The aim of this paper was to analyze these branching patterns in human cerebral cortex. Digital three dimensional images of the microvascular network were obtained from thick sections of India ink-injected human brain by confocal laser microscopy covering a large zone of secondary cortex. A novel segmentation method was used to extract the skeletons of 228 vascular trees (152 arterioles and 76 venules) and measure the diameter at every vertex. The branching patterns (area ratios and angles of bifurcations) of nearly 10,000 bifurcations of cortical vascular trees were analyzed, establishing their statistical properties and structural variations as a function of the vessel nature (arterioles versus venules), the parent vessel topological order or the bifurcation type. We also describe their connectivity and discuss the relevance of the assumed optimal design of vascular branching to account for the complex nature of microvascular architecture. The functional implications of some of these structural variations are considered. The branching patterns established from a large database of a human organ contributes to a better understanding of the bifurcation design and provides an essential reference both for diagnosis and for a future large reconstruction of cerebral microvascular network. PMID- 20005217 TI - Next generation OP-bioscavengers: a circulatory long-lived 4-PEG hypolysine mutant of F338A-HuAChE with optimal pharmacokinetics and pseudo-catalytic characteristics. AB - We have shown previously that conjugation of polyethylene glycol (PEG) chains to recombinant human acetylcholinesterase (rHuAChE) results in the extension of its residence time in the circulation of mice and monkeys [1,2]. By profiling the pharmacokinetic behavior of an array of well-defined hypolysine human mutant AChE molecules following PEGylation, we now determine that the duration of these enzyme forms in the circulation of rhesus macaques correlates with their number of appended PEG moieties, and is influenced by the actual location of the PEG chains at the molecule surface, as well. These findings, which concur with those we have previously established in mice, indicate that a common set of rules dictates the circulatory fate of PEGylated HuAChEs in rodents and non-human primates. In addition to its effect on circulatory residence, PEGylation reduces the ability of the rHuAChE bioscavenger to elicit an immune response in the heterologous mouse animal system. Thus, an inverse relationship between anti-AChE antibody production and PEG loading was observed following repeated administration of the different PEGylated hypolysine human AChEs to mice. We note however, that in rhesus macaques, the essentially homologous (human) AChE does not induce specific anti-AChE antibodies after repeated administration of high doses of the enzyme in its PEGylated form, and even in its non-PEGylated form. Taken together, these findings indicate that PEG acts by veiling enzyme-related epitopes, which would otherwise interact with host circulatory elimination pathways and immune system. The barring of such interactions by obstructive PEGs, confers the enzyme molecule with both extended circulatory residence and mitigated immunogenic properties. Further modulation by incorporation of the F338A mutation into the PEGylated hypolysine rHuAChE enzyme mold, resulted in the generation of an OP-bioscavenger that displayed reduced aging rates and could effectively protect mice against repeated exposure to CW agents. This selected 4 PEG F338A-AChE can serve as a paradigm for new generation OP-bioscavengers, specifically tailored for prophylactic treatment against toxic OP-agents. PMID- 20005218 TI - A potential regulatory single nucleotide polymorphism in the promoter of the Klotho gene may be associated with essential hypertension in the Chinese Han population. AB - BACKGROUND: Mice with defects in the Klotho gene exhibit multiple aging phenotypes including arteriosclerosis. We hypothesised that the G-395A polymorphism in the promoter region of the human Klotho gene may contribute to the prevalence of Essential Hypertension (EH). METHODS: We investigate whether the G-395A polymorphism of Klotho is associated with EH in a population consisting of 215 patients with EH and 220 non-hypertensive subjects. We also tested whether a G/A substitution at the G-395A site affected the transcription level in vitro through the dual-luciferase reporter assay. RESULTS: Differences in the genotype distributions of the G-395A polymorphism between the EH and non hypertension groups are statistically significant (P=0.032). There are differential effects of age, gender and smoking status on the association of the G-395A polymorphism with EH; the G-395A polymorphism is significantly associated with EH in subjects over 60years old, in females and in nonsmokers. A multiple logistic regression analysis indicated that the odds ratio for EH in the -395A allele carriers as compared with the control group was 0.593 (P=0.024) after adjusting for current traditional risk factors. The dual-luciferase reporter assay revealed that the -395A carrier of a 498-bp DNA fragment (containing the G 395A site) upstream of the Klotho gene has higher relative luciferase activity than the -395G carrier. CONCLUSIONS: The G-395A polymorphism of the human Klotho gene is associated with EH and may be a potential regulatory site. PMID- 20005219 TI - Nlcam modulates midline convergence during anterior neural plate morphogenesis. AB - During development, different cell types must undergo distinct morphogenetic programs so that tissues develop the right dimensions in the appropriate place. In early eye morphogenesis, retinal progenitor cells (RPCs) move first towards the midline, before turning around to migrate out into the evaginating optic vesicles. Neighbouring forebrain cells, however, converge rapidly and then remain at the midline. These differential behaviours are regulated by the transcription factor Rx3. Here, we identify a downstream target of Rx3, the Ig-domain protein Nlcam, and characterise its role in regulating cell migration during the initial phase of optic vesicle morphogenesis. Through sophisticated live imaging and comprehensive cell tracking experiments in zebrafish, we show that ectopic expression of Nlcam in RPCs, as is observed in Rx3 mutants, causes enhanced convergence of these cells. Expression levels of Nlcam therefore regulate the migratory properties of RPCs. Our results provide evidence that the two phases of optic vesicle morphogenesis: slowed convergence and outward-directed migration, are under different genetic control. We propose that Nlcam forms part of the guidance machinery directing rapid midline migration of forebrain precursors, where it is normally expressed, and that its ectopic expression upon loss of Rx3 imparts these migratory characteristics upon RPCs. PMID- 20005220 TI - Autologous bone marrow mesenchymal cell transplantation improves left ventricular function in a rabbit model of dilated cardiomyopathy. AB - BACKGROUND: Bone marrow mesenchymal stem cells (BMSCs) could differentiate into cardiomyocytes. In this study, we wished to determine whether we could improve ventricular function in a dilated cardiomyopathy (DCM) model in the rabbit by injecting stem cells directly into the ventricular wall. METHODS AND RESULTS: Japanese rabbits were treated with doxorubicin to induce dilated cardiomyopathy. BMSCs were isolated from isogenic adult rabbits and expanded ex vivo. DCM-model animals received an injection directly into the ventricular wall with either BMSCs or saline (sham-operated control; a third control group underwent no operation). Heart function in all three groups was evaluated by echocardiography and cardiac catheterization. Hearts were later excised for histological and immunostaining analysis. Left ventricular function was significantly better in BMSC-transplanted animals than in controls. Cellular staining of BrdU and N cadherin, indicating the presence of bone-marrow-derived cells, was found in BMSC transplanted animals but not in control and sham-operated animals. CONCLUSIONS: BMSC transplantation can improve left ventricular function in DCM. Direct injection into the ventricular wall is an effective approach for stem cell delivery. PMID- 20005221 TI - Hypoxia and the expression of HIF-1alpha and HIF-2alpha in the retina of streptozotocin-injected mice and rats. AB - Decreases in retinal blood flow in diabetics could render the retina hypoxic. In mouse and rat models of diabetes, a decrease in retinal blood flow occurs early, within 3-4 weeks of the induction of hyperglycemia, although information is scarce on whether this early decrease in flow induces hypoxia. The purpose of the current study was to determine whether hypoxia-inducible factor (HIF) levels increase following 4 and/or 12 weeks of hyperglycemia in streptozotocin (STZ) injected mouse (C57BL/6) and rat (Wistar) retinas. Additionally, retinal tissue hypoxia was measured with pimonidazole following 12 weeks of hyperglycemia. These aims were accomplished via immunostaining of cross-sections from enucleated eyes. In mice, staining for HIF-1alpha and HIF-2alpha showed a contrasting pattern, with HIF-1alpha higher in the inner retina than outer, but HIF-2alpha higher in the outer retina than inner. However, in rats, staining for both HIF-1alpha and HIF-2alpha was more intense in the inner retina. The HIF-1alpha staining intensities and patterns were similar between diabetic animals and their non diabetic counterparts following 4 and 12 weeks of hyperglycemia. The same was true for HIF-2alpha except for a trend toward an increase following 12 weeks of hyperglycemia in mice. Pimonidazole staining showed significant decreases throughout all layers of the central retina and most layers of the peripheral retina of rats (but not mice), following 12 weeks of hyperglycemia. In summary, despite early decreases in flow in rats and mice, retinal HIF-1alpha and HIF 2alpha were not found to be increased, and the extent of hypoxia may even decrease after 12 weeks of hyperglycemia in rats. PMID- 20005222 TI - Proinflammatory chemokines during Candida albicans keratitis. AB - Chemotactic cytokines mediate the recruitment of leukocytes into infected tissues. This study investigated the profile of chemokines during experimental Candida albicans keratitis and determined the effects of chemokine inhibition on leukocyte infiltration and fungal growth during murine keratomycosis. Scarified corneas of BALB/c mice were topically inoculated with C. albicans and monitored daily over one week for fungal keratitis. After a gene microarray for murine chemokines compared infected corneas to controls, real-time reverse transcription polymerase chain reaction (RT-PCR) and immunostaining assessed chemokine expression in infected and mock-inoculated corneas. An anti-chemokine antibody was then administered subconjunctivally and evaluated for effects on clinical severity, corneal inflammation, fungal recovery, and cytokine expression. Of 33 chemokine genes examined by microarray, 6 CC chemokines and 6 CXC chemokines were significantly (P<0.05) upregulated more than two-fold. Chemokine (CC-motif) ligand 3 (CCL3) was upregulated 108-fold (P=0.03) by real-time RT-PCR within one day after fungal inoculation and remained increased 28-fold (P=0.02) at one week, and its in situ expression increased in the epithelium and stroma of infected corneas. Compared to the control antibody-treated group, eyes treated with anti CCL3 antibody showed reduced clinical severity (P<0.05), less corneal neovascularization (P=0.02), and fewer inflammatory cells infiltrating corneal tissue, but the amount of recoverable fungi was not significantly (P=0.4) affected. Anti-CCL3 treatment significantly (P=0.01) reduced the expression of tumor necrosis factor and interleukin-1beta in infected corneas. These results indicate that chemokines, especially the CC chemokine CCL3, play important roles in the acute inflammatory response to C. albicans corneal infection. PMID- 20005224 TI - Perpendicular organization of sympathetic neurons within a required physiological voltage. AB - In vitro, postganglionic sympathetic neurons (PSNs) profoundly organize their anatomy according to cues provided by an extracellular voltage. Over 90% of PSNs retract neurites that are parallel/tangential to a gradient of approximately 400 mV/mm. Complete neurite retraction takes approximately 20-40 minutes. Subsequently, neurites grow out from the soma, but now perpendicular to the lines of force while branching profusely. The complete restructuring of the neurons anatomy takes 2-3 hours at 35 degrees C. The maintenance of this asymmetrical anatomy requires the continuous presence of the extracellular electrical field (Ef). We discuss this observation relative to the organization of neurons residing in natural voltage gradients that exist across all epithelia in which neurons are born, mature, or migrate. PMID- 20005223 TI - Acute glial activation by stab injuries does not lead to overt damage or motor neuron degeneration in the G93A mutant SOD1 rat model of amyotrophic lateral sclerosis. AB - Amyotrophic lateral sclerosis (ALS) is a neurodegenerative disease where motor neurons within the brain and spinal cord are lost, leading to paralysis and death. Recently, a correlation between head trauma and the incidence of ALS has been reported. Furthermore, new invasive neurosurgical studies are being planned which involve inserting needles directly to the spinal cord. We therefore tested whether acute trauma to the spinal cord via a knife wound injury would lead to accelerated disease progression in rodent models of ALS (SOD1(G93A) rats). A longitudinal stab injury using a small knife was performed within the lumbar spinal cord region of presymptomatic SOD1(G93A) rats. Host glial activation was detected in the lumbar area surrounding a micro-knife lesion at 2 weeks after surgery in both wild type and SOD1(G93A) animals. However, there was no sign of motor neuron loss in the injured spinal cord of any animal and normal motor function was maintained in the ipsilateral limb. These results indicate that motor neurons in presymptomatic G93A animals are not affected by an invasive puncture wound injury involving reactive astrocytes. Furthermore, acute trauma alone does not accelerate disease onset or progression in this ALS model which is important for future strategies of gene and cell therapies directly targeting the spinal cord of ALS patients. PMID- 20005225 TI - An unbiased, staged, multicentre, validation strategy for Alzheimer's disease CSF tau levels. AB - Newly proposed diagnostic criteria for Alzheimer's disease include cerebrospinal fluid (CSF) tau levels as one core supportive criterion. The published high sensitivity and specificity figures for CSF tau levels in Alzheimer's disease are offset by the large range of proposed cutoff values (9.6 pg/mL to 1140 pg/mL). This study aimed to provide guidance on how to establish, validate and audit CSF tau cutoff values using an unbiased, two-stage multicentre strategy. Both receiver operator characteristics (ROC) optimised and population-based cutoff values were calculated on a pilot dataset (n=99), validated in a large dataset (n=560) and then compared to the literature. The data suggest using an ROC optimised cutoff level of 323+/-51.7 pg/mL allowing for the published inter laboratory coefficient of variation of 16%. This cutoff level was confirmed in a prospective audit (n=100). As demand for CSF tau levels will increase globally, the accuracy of local CSF hTau cutoff levels can be compared against this benchmark. PMID- 20005226 TI - A role of Histidine151 in the lamprey gonadotropin-releasing hormone receptor-1 (lGnRHR-1): Functional insight of diverse amino acid residues in the position of Tyr of the DRY motif in GnRHR from an ancestral type II receptor. AB - The highly conserved DRY motif located at the end of the third transmembrane of G protein-coupled receptors has been described as a key motif for several aspects of GPCR functions. However, in the case of the vertebrate gonadotropin-releasing hormone receptor (GnRHR), the amino acid in the third position in the DRY motif is variable. In the lamprey, a most basal vertebrate, the third amino acid of the "DRY" in lamprey (lGnRHR-1) is His, while it is most often His/Gln in the type II GnRHR. To investigate the functional significance of the substitution of DRY to DRH in the GnRHR-1, second messenger signaling, ligand binding and internalization of the wild-type and mutant lGnRH receptors were characterized with site-directed mutagenesis. Treatment of the DRE(151) and DRS(151) mutant receptors with lamprey GnRH-I significantly reduced inositol phosphate compared to wild-type (DRH(151)) and DRY(151) receptors. The LogIC(50) of wild-type receptor (-9.554+/-0.049) was similar to the LogIC(50) of DRE(151), DRS(151) and DRX(151) mutants, yet these same mutants were shown to significantly reduce cell surface expression. However, the DRY(151) mutant compared to the wild-type receptor increased cell-surface expression, suggesting that the reduction of IP production was due to the level of the cell-surface expression of the mutant receptors. The rate of internalization of DRX(151) (35.60%) was reduced compared to wild-type and other mutant receptors. These results suggest that His(151) of the lamprey GnRH receptor-1 may play a critical role in the retention of a certain level of cell-surface expression for subsequent cellular second messenger events. PMID- 20005227 TI - Ghrelin: an emerging player in the regulation of reproduction in non-mammalian vertebrates. AB - The endocrine regulation of vertebrate reproduction is achieved by the coordinated actions of multiple endocrine factors mainly produced from the brain, pituitary, and gonads. In addition to these, several other tissues including the fat and gut produce factors that have reproductive effects. Ghrelin is one such gut/brain hormone with species-specific effects in the regulation of mammalian reproduction. Recent studies have shown that ghrelin and ghrelin receptor mRNAs, and protein are expressed in the ovary and testis of mammals, indicating a direct effect for ghrelin in the control of reproduction. Ghrelin regulates mammalian reproduction by modulating hormone secretion from the brain and pituitary, and by acting directly on the gonads to influence reproductive tissue development and steroid hormone release. Based on the studies reported so far, ghrelin seems to have a predominantly inhibitory role on mammalian reproduction. The presence of ghrelin and ghrelin receptor has been found in the brain, pituitary and gonads of several non-mammalian vertebrates. In contrast to mammals, ghrelin seems to have a stimulatory role in the regulation of non-mammalian reproduction. The main objective of this review is to do a perspective analysis of the comparative aspects of ghrelin regulation of reproduction. PMID- 20005228 TI - Relationship between alpha-melanocyte-stimulating hormone- and neuropeptide Y containing neurons in the goldfish hypothalamus. AB - Intracerebroventricular (ICV) injection of alpha-melanocyte-stimulating hormone (alpha-MSH) inhibits, whereas ICV injection of neuropeptide Y (NPY) stimulates food intake in the goldfish. However, there is little information about the functional relationship between alpha-MSH-induced anorexigenic and NPY-induced orexigenic actions in the goldfish. In this study we examined the relationship between alpha-MSH- and NPY-containing neurons in the goldfish hypothalamus to investigate whether these alpha-MSH- and NPY-containing neurons have direct mutual inputs. alpha-MSH- and NPY-like immunoreactivities were distributed throughout the brain, especially in the diencephalon. In particular, alpha-MSH containing nerve fibers or endings lay in close apposition to NPY-containing neurons in a specific region of the hypothalamus, the nucleus posterioris periventricularis (NPPv). NPY-containing nerve fibers or endings also lay in close apposition to alpha-MSH-containing neurons specifically in the interior part of the nucleus lateralis tuberis (NLTi). We also investigated the effect of ICV injection of melanocortin 4 receptor agonist (melanotan II) at 100 pmol/g body weight (BW), which is enough to suppress food intake, or NPY at 10 pmol/g BW, which is enough to enhance food intake, on expression levels of mRNA for NPY or proopiomelanocortin (POMC) in the hypothalamus. ICV injection of melanotan II and NPY induced a significant decrease in the expression levels for NPY and POMC mRNA, respectively. These observations suggest that alpha-MSH- and NPY-containing neurons share direct mutual inputs in the NPPv and the NLTi of the hypothalamus, and that alpha-MSH and NPY functionally interact or exhibit mutual inhibition to regulate feeding behavior in the goldfish. PMID- 20005229 TI - Conserved genetic pathways controlling the development of the diffuse endocrine system in vertebrates and Drosophila. AB - The midgut epithelium is formed by absorptive enterocytes, secretory cells and endocrine cells. Each of these lineages is derived from the pluripotent progenitors that constitute the embryonic endoderm; the mature midgut retains pools of self-renewing stem cells that continue to produce all lineages. Recent findings in vertebrates and Drosophila shed light on the genetic mechanism that specifies the fate of the different lineages. A pivotal role is played by the Notch signaling pathway that, in a manner that appears to be very similar to the way in which Notch signaling selects neural progenitors within the neurectoderm, distinguishes the fate of secretory/endocrine cells and enterocytes. Proneural genes encoding bHLH transcription factors are expressed and required in prospective endocrine cells; activation of the Notch pathways restricts the number of these cells and promotes enterocyte development. In this review we compare the development of the intestinal endocrine cells in vertebrates and insects and summarize recent findings dealing with genetic pathways controlling this cell type. PMID- 20005230 TI - Plasma ghrelin concentrations change with physiological state in a sciurid hibernator (Spermophilus lateralis). AB - Ghrelin is a recently discovered hormone which has profound effects on food intake and lipogenesis in mammals. In all mammals studied thus far, plasma ghrelin concentrations are increased before a meal and decrease immediately following a meal; ghrelin levels increase with fasting. The golden-mantled ground squirrel Spermophilus lateralis (also known as Callospermophilus lateralis (see Helgen et al., 2009) is a diurnal hibernator which has a robust annual cycle of body mass gain and loss that is primarily controlled by food intake. We hypothesized that in spring, summer, and autumn, the endogenous ghrelin concentrations of hibernators would be similar to those of non-hibernators, but that during the winter hibernation season, plasma ghrelin concentrations would be low or undetectable. We found that peripherally injected ghrelin significantly increased food intake in June. Plasma ghrelin concentrations were significantly increased through 5 days of fasting during a short-term fast in summer. Over a 24h period, ghrelin concentrations increased at night and decreased during the day with drops corresponding to times when squirrels were eating. In January, ghrelin concentrations are low but measurable even while animals are at low body temperature (Tb). This is the first report of ghrelin concentrations in a non photoperiodic hibernator. We suggest that ghrelin may be important for the regulation of food intake and the body mass cycle in mammals that hibernate. PMID- 20005231 TI - Reproductive axis function and gonadotropin microheterogeneity in a male rat model of diet-induced obesity. AB - Obesity causes complex metabolic and endocrine changes that may lead to adverse outcomes, including hypogonadism. We herein studied the reproductive axis function in male rats under a high-fat diet and analyzed the impact of changes in glycosylation of pituitary LH on the bioactivity of this gonadotropin. Rats were fed with a diet enriched in saturated fat (20% of total calories) and euthanized on days 90 or 180 of diet. Long-term (180 days), high-fat feeding rats exhibited a metabolic profile compatible with insulin resistance and metabolic syndrome; they concomitantly showed decreased intrapituitary and serum LH concentrations, low serum testosterone levels, and elevated serum 17beta-estradiol concentrations. A fall in biological to immunological ratio of intrapituitary LH was detected in 180 days control diet-treated rats but not in high-fat-fed animals, as assessed by a homologous in vitro bioassay. Chromatofocusing of pituitary extracts yielded multiple LH charge isoforms; a trend towards decreased abundance of more basic isoforms (pH 9.99-9.0) was apparent in rats fed with the control diet for 180 days but not in those that were fed the diet enriched in saturated fat. It is concluded that long-term high-fat feeding alters the function of the pituitary-testicular axis, resulting in hypogonadotropic hypogonadism. The alterations in LH function found in these animals might be subserved by changes in hypothalamic GnRH output and/or sustained gonadotrope exposure to an altered sex steroid hormone milieu, representing a distinctly different regulatory mechanism whereby the pituitary attempts to counterbalance the effects of long-term obesity on reproductive function. PMID- 20005232 TI - Changes in abundance of aquaporin-like proteins occurs concomitantly with seasonal acquisition of freeze tolerance in the goldenrod gall fly, Eurosta solidaginis. AB - The accumulation of cryoprotectants and the redistribution of water between body compartments play central roles in the capacity of insects to survive freezing. Aquaporins (AQPs) allow for rapid redistribution of water and small solutes (e.g. glycerol) across the cell membrane and were recently implicated in promoting freeze tolerance. Here, we examined whether aquaporin-like protein abundance correlated with the seasonal acquisition of freezing tolerance in the goldenrod gall fly, Eurosta solidaginis (Diptera: Tephritidae). Through the autumn, larvae became tolerant of freezing at progressively lower temperatures and accumulated the cryoprotectant glycerol. Furthermore, larvae significantly increased the abundance of membrane-bound aquaporin and aquaglyceroporin-like proteins from July through January. Acute exposure of larvae to cold and desiccation resulted in upregulation of the AQP3-like proteins in October, suggesting that their abundance is regulated by environmental cues. The seasonal increase in abundance of both putative aquaporins and aquaglyceroporins supports the hypothesis that these proteins are closely tied to the seasonal acquisition of freeze tolerance, functioning to permit cells to quickly lose water and take-up glycerol during extracellular ice formation, as well as reestablish water and glycerol concentrations upon thawing. PMID- 20005233 TI - Skin deep: what can the study of dermal fibroblasts teach us about dilated cardiomyopathy? PMID- 20005235 TI - A model for Ca2+ waves in networks of glial cells incorporating both intercellular and extracellular communication pathways. AB - Networks of glial cells, and in particular astrocytes, are capable of sustaining calcium (Ca(2+)) waves both in vivo and in vitro. Experimentally, it has been shown that there are two separate modes of communication: the first by the passage of an agent (inositol 1,4,5-triphosphate, IP(3)) through gap junctions (GJs) joining cells; the second by the diffusion of an extracellular agent (adenosine triphosphate, ATP) that binds to receptors on the cells. In both cases, the outcome is the release of Ca(2+) from internal stores in the glial cells. These two modes of communication are not mutually exclusive, but probably work in conjunction in many cases. We present a model of a two-dimensional network of glial cells that incorporates regenerative intercellular (GJ) and extracellular (ATP) pathways. In the extreme cases of only one type of pathway, the results are in agreement with previous models. Adding an extracellular pathway to the GJ model increased the extent and duration of the Ca(2+) wave, but did not significantly change the speed of propagation. Conversely, adding GJs to the extracellular model did increase the wave speed. The model was modified to apply to the retina by extending it to include both astrocytes and Muller cells, with GJs the dominant coupling between astrocytes and ATP responsible for most of the remaining communication. It was found that both pathways are necessary to account for experimental results. PMID- 20005234 TI - The electrostatics of VDAC: implications for selectivity and gating. AB - The voltage-dependent anion channel (VDAC) is the major pathway mediating the transfer of metabolites and ions across the mitochondrial outer membrane. Two hallmarks of the channel in the open state are high metabolite flux and anion selectivity, while the partially closed state blocks metabolites and is cation selective. Here we report the results from electrostatics calculations carried out on the recently determined high-resolution structure of murine VDAC1 (mVDAC1). Poisson-Boltzmann calculations show that the ion transfer free energy through the channel is favorable for anions, suggesting that mVDAC1 represents the open state. This claim is buttressed by Poisson-Nernst-Planck calculations that predict a high single-channel conductance indicative of the open state and an anion selectivity of 1.75--nearly a twofold selectivity for anions over cations. These calculations were repeated on mutant channels and gave selectivity changes in accord with experimental observations. We were then able to engineer an in silico mutant channel with three point mutations that converted mVDAC1 into a channel with a preference for cations. Finally, we investigated two proposals for how the channel gates between the open and the closed state. Both models involve the movement of the N-terminal helix, but neither motion produced the observed voltage sensitivity, nor did either model result in a cation-selective channel, which is observed experimentally. Thus, we were able to rule out certain models for channel gating, but the true motion has yet to be determined. PMID- 20005236 TI - A competition model between Pseudomonas fluorescens and pathogens via iron chelation. AB - In this study we present a competition model between a non-chelator (e.g. pathogen) microorganism and an iron chelator microorganism (e.g. Pseudomonas fluorescens). This latter is a beneficial bacteria that can inhibit the growth of the non-chelator through its iron chelating capability. This phenomena of iron chelation is shown to prevent the pathogen from proliferating to numbers capable of causing disease. A mathematical model is formulated and used to study this competition. The model proposes a new and simple conceptual explanation of interactions. It is a nonlinear system of ordinary differential equations. A qualitative analysis of the model for the batch case (no inflow or outflow from the system) is carried out and the global behavior of the model variables is studied. For the chemostat case, the equilibrium points were derived and their stability was performed through extensive numerical simulations. It is found that iron chelation is able to control the non-chelator microorganism growth under a wide range of conditions. PMID- 20005237 TI - Functional and biochemical evidence indicating beneficial effect of Melatonin and Nicotinamide alone and in combination in experimental diabetic neuropathy. AB - Oxidative stress resulting in excessive generation of ROS is a compelling initiator of DNA damage along with damage to various cellular proteins and other macromolecules. Poly(ADP-ribose) polymerase (PARP) activation in response to DNA damage, stirs an energy-consuming cellular metabolic cycle; culminating into cell death. The present study was designed to determine the effect of combining an antioxidant, Melatonin and a PARP inhibitor, Nicotinamide on the hallmark deficits developing in diabetic neuropathy (DN). Streptozotocin (STZ, 55 mg/kg, i.p.) was administered to induce diabetes. Six weeks post diabetes induction, two week treatment with Melatonin (3 and 10 mg/kg) and Nicotinamide (100 and 300 mg/kg) either alone or in combination was given. Effect of these interventions on the functional, behavioral and biochemical changes caused by hyperglycemia were studied in treated animals. Melatonin and Nicotinamide alone as well as in combination ameliorated the functional deficits along with improvement in pain parameters. The combination also demonstrated an essential reversal of biochemical alterations. Nitrotyrosine and Poly ADP Ribose (PAR) immunopositivity was significantly decreased in sciatic nerve micro-sections of treatment group. The results of this study advocate that simultaneous inhibition of oxidative stress-PARP activation cascade may prove useful for the pharmacotherapy of DN. PMID- 20005238 TI - Atypical hemispheric asymmetry in the perception of negative human vocalizations in individuals with Williams syndrome. AB - Williams syndrome is a neurological condition associated with high levels of auditory reactivity and emotional expression combined with impaired perception of prosody. Yet, little is currently known about the neural organization of affective auditory processing in individuals with this disorder. The current study examines auditory emotion processing in individuals with Williams syndrome. Hemispheric organization for positive and negative human non-linguistic sound processing was compared in participants with and without the disorder using a dichotic listening paradigm. While controls exhibited an expected right cerebral hemisphere advantage for processing negative sounds, those with Williams syndrome showed the opposite pattern. No differences between the groups emerged for the positive stimuli. The results suggest aberrant processing of negative auditory information in Williams syndrome. PMID- 20005239 TI - Recognition of emotional prosody is altered after subthalamic nucleus deep brain stimulation in Parkinson's disease. AB - The recognition of facial emotions is impaired following subthalamic nucleus (STN) deep brain stimulation (DBS) in Parkinson's disease (PD). These changes have been linked to a disturbance in the STN's limbic territory, which is thought to be involved in emotional processing. This was confirmed by a recent PET study where these emotional modifications were correlated with changes in glucose metabolism in different brain regions, including the amygdala and the orbitofrontal regions that are well known for their involvement in emotional processing. Nevertheless, the question as to whether these emotional changes induced by STN DBS in PD are modality-specific has yet to be answered. The objective of this study was therefore to examine the effects of STN DBS in PD on the recognition of emotional prosody. An original emotional prosody paradigm was administered to twenty-one post-operative PD patients, twenty-one pre-operative PD patients and twenty-one matched controls. Results showed that both the pre- and post-operative groups differed from the healthy controls. There was also a significant difference between the pre and post groups. More specifically, an analysis of their continuous judgments revealed that the performance of the post operative group compared with that of the other two groups was characterized by a systematic emotional bias whereby they perceived emotions more strongly. These results suggest that the impaired recognition of emotions may not be specific to the visual modality but may also be present when emotions are expressed through the human voice, implying the involvement of the STN in the brain network underlying the recognition of emotional prosody. PMID- 20005240 TI - Just another face in the crowd: evidence for decreased detection of angry faces in children with Williams syndrome. AB - The detection of social threat is crucial for adaptive behaviour. Previous studies have shown that angry faces capture attention and are processed more efficiently than happy faces. While this anger superiority effect has been found in typical and atypical development, it is unknown whether it exists in individuals with Williams syndrome (WS), who show reduced social fear and atypical sociability. In this study, children with WS searched for angry or happy target faces surrounded by 2, 5 or 8 distracters (happy or angry faces, respectively). Performance was compared to that of mental age-matched controls. Results revealed no group differences for happy faces, however for angry faces, the WS, but not the control group, showed a significant performance decrease for the 8-distracters condition, indicating the absence of an anger superiority effect, in good agreement with evidence for abnormal structure and function in brain areas for social threat processing in WS. PMID- 20005241 TI - F(ST) in the cytonuclear system. AB - Selection on nuclear (or organelle) sites inevitably affects the spatial distribution of a neutral organelle (or nuclear) allele via transient cytonuclear disequilibrium. Here I examine this effect in terms of F(st) for a neutral allele by bringing together cytonuclear genomes with contrasting modes of inheritance. The relationships between cytonuclear disequilibrium and increment in F(st) are explored and confirmed through Monte Carlo simulations. Results show that the transient increment in F(st) for a neutral allele is not only related to the vectors of seed and pollen dispersal but also to the mode of its inheritance. Such increments can be substantial under certain conditions. Seed dispersal is more effective than pollen dispersal in changing the transient increment. The cumulative effects from multiple selective nuclear sites can amplify the transient increment in F(st) for a neutral paternal or maternal organelle allele. Selection on selective organelle sites facilitates the transient increment in F(st) for a neutral nuclear allele. Partial selfing can significantly reinforce the transient increment in F(st). These theoretical insights highlight the roles of transient cytonuclear disequilibrium as a biological factor in evolving population differentiation and refine our practical interpretations of F(st) with cytonuclear markers. PMID- 20005242 TI - ATP7B detoxifies silver in ciliated airway epithelial cells. AB - Silver is a centuries-old antibiotic agent currently used to treat infected burns. The sensitivity of a wide range of drug-resistant microorganisms to silver killing suggests that it may be useful for treating refractory lung infections. Toward this goal, we previously developed a methylated caffeine silver acetate compound, SCC1, that exhibits broad-spectrum antimicrobial activity against clinical strains of bacteria in vitro and when nebulized to lungs in mouse infection models. Preclinical testing of high concentrations of SCC1 in primary culture mouse tracheal epithelial cells (mTEC) showed selective ciliated cell death. Ciliated cell death was induced by both silver- and copper-containing compounds but not by the methylated caffeine portion of SCC1. We hypothesized that copper transporting P-type ATPases, ATP7A and ATP7B, play a role in silver detoxification in the airway. In mTEC, ATP7A was expressed in non-ciliated cells, whereas ATP7B was expressed only in ciliated cells. The exposure of mTEC to SCC1 induced the trafficking of ATP7B, but not ATP7A, suggesting the presence of a cell-specific silver uptake and detoxification mechanisms. Indeed, the expression of the copper uptake protein CTR1 was also restricted to ciliated cells. A role of ATP7B in silver detoxification was further substantiated when treatment of SCC1 significantly increased cell death in ATP7B shRNA-treated HepG2 cells. In addition, mTEC from ATP7B(-/-) mice showed enhanced loss of ciliated cells compared to wild type. These studies are the first to demonstrate a cell type specific expression of the Ag+/Cu+ transporters ATP7A, ATP7B, and CTR1 in airway epithelial cells and a role for ATP7B in detoxification of these metals in the lung. PMID- 20005243 TI - Accessing novel conoidean venoms: Biodiverse lumun-lumun marine communities, an untapped biological and toxinological resource. AB - Cone snail venoms have yielded pharmacologically active natural products of exceptional scientific interest. However, cone snails are a small minority of venomous molluscan biodiversity, the vast majority being tiny venomous morphospecies in the family Turridae. A novel method called lumun-lumun opens access to these micromolluscs and their venoms. Old fishing nets are anchored to the sea bottom for a period of 1-6months and marine biotas rich in small molluscs are established. In a single lumun-lumun community, we found a remarkable gastropod biodiversity (155 morphospecies). Venomous predators belonging to the superfamily Conoidea (36 morphospecies) were the largest group, the majority being micromolluscs in the family Turridae. We carried out an initial analysis of the most abundant of the turrid morphospecies recovered, Clathurella (Lienardia) cincta (Dunker, 1871). In contrast to all cDNA clones characterized from cone snail venom ducts, one of the C. cincta clones identified encoded two different peptide precursors presumably translated from a single mRNA. The prospect of easily accessing so many different morphospecies of venomous marine snails raises intriguing toxinological possibilities: the 36 conoidean morphospecies in this one net alone have the potential to yield thousands of novel pharmacologically active compounds. PMID- 20005244 TI - The relationship between lens transmission and opsin gene expression in cichlids from Lake Malawi. AB - The lens plays an important role in regulating the wavelengths of light that reach the retina. However, the evolutionary relationship between lens transmission and retinal sensitivity remains cloudy at best. We examined the relationship between lens transmission and opsin gene expression in a group of rapidly radiating cichlids from East Africa. Lens transmission was bimodal, either cutting off around 360 or 400 nm, and appeared to be quite labile evolutionarily. We found a strong correlation between lens transmission and SWS1 (UV) opsin gene expression, suggesting that UV transmitting lenses are adaptive in cichlids. Species which expressed high levels of SWS2B (violet) opsin varied in their lens transmission while most species that expressed high levels of SWS2A (blue) opsin had UV blocking lenses. In no instance did lens transmission appear to limit retinal sensitivity. Finally, the strong correlation that we observe between SWS1 expression and lens transmission suggests that these two traits might be coupled genetically. PMID- 20005245 TI - An empirical comparative study on biological age estimation algorithms with an application of Work Ability Index (WAI). AB - In this study, we described the characteristics of five different biological age (BA) estimation algorithms, including (i) multiple linear regression, (ii) principal component analysis, and somewhat unique methods developed by (iii) Hochschild, (iv) Klemera and Doubal, and (v) a variant of Klemera and Doubal's method. The objective of this study is to find the most appropriate method of BA estimation by examining the association between Work Ability Index (WAI) and the differences of each algorithm's estimates from chronological age (CA). The WAI was found to be a measure that reflects an individual's current health status rather than the deterioration caused by a serious dependency with the age. Experiments were conducted on 200 Korean male participants using a BA estimation system developed principally under the concept of non-invasive, simple to operate and human function-based. Using the empirical data, BA estimation as well as various analyses including correlation analysis and discriminant function analysis was performed. As a result, it had been confirmed by the empirical data that Klemera and Doubal's method with uncorrelated variables from principal component analysis produces relatively reliable and acceptable BA estimates. PMID- 20005246 TI - Perspectives on protein crystallisation. AB - This final part on 'perspectives' is focused on new strategies that can be used to crystallise proteins and improve the crystal quality of macromolecular complexes using any of the methods reviewed in this focused issue. Some advantages and disadvantages, limitations, and plausible applications to high resolution X-ray crystallography are discussed. PMID- 20005247 TI - Crystal growth of proteins, nucleic acids, and viruses in gels. AB - Medium-sized single crystals with perfect habits and no defect producing intense and well-resolved diffraction patterns are the dream of every protein crystallographer. Crystals of biological macromolecules possessing these characteristics can be prepared within a medium in which mass transport is restricted to diffusion. Chemical gels (like polysiloxane) and physical gels (such as agarose) provide such an environment and are therefore suitable for the crystallisation of biological macromolecules. Instructions for the preparation of each type of gel are given to urge crystal growers to apply diffusive media for enhancing crystallographic quality of their crystals. Examples of quality enhancement achieved with silica and agarose gels are given. Results obtained with other substances forming gel-like media (such as lipidic phases and cellulose derivatives) are presented. Finally, the use of gels in combination with capillary tubes for counter-diffusion experiments is discussed. Methods and techniques implemented with proteins can also be applied to nucleic acids and nucleoprotein assemblies such as viruses. PMID- 20005248 TI - Are those who use specific complementary and alternative medicine therapies less likely to be immunized? AB - OBJECTIVE: Some authorities are concerned that the use of complementary and alternative medications (CAM) may replace recommended preventive health practices. This study was done to determine if users of individual types of CAM were less likely to receive recommended immunizations. METHODS: We used data from the 2007 National Health Interview Survey of over 23,000 adult, non institutionalized U.S. citizens using bivariate and multivariate analysis to determine if users of individual types of CAM were less likely to receive influenza and/or pneumococcal vaccinations. RESULTS: Using a weighted logistic regression analysis, we found that respondents who used chiropractic care were less likely to receive flu shots (OR=0.68, CI=0.55,0.83, p<0.001). There was a mildly positive trend toward receiving the pneumococcal vaccine in users of deep breathing exercises and toward not receiving both in followers of qi gong. Prayer use was prevalent and had a positive impact on receiving immunizations, especially in Blacks and those in poor health. Regular exercise, having a primary care provider and more frequent office visits were also positively associated with receiving immunizations. CONCLUSION: Chiropractic users are less likely to get flu shots, perhaps reflecting their national body's attitude, which could affect morbidity and mortality. Providers should be aware of their patients' CAM use and encourage accepted primary care practices. PMID- 20005249 TI - Event-related brain potential correlates of visual awareness. AB - Electrophysiological recordings during visual tasks can shed light on the temporal dynamics of the subjective experience of seeing, visual awareness. This paper reviews studies on electrophysiological correlates of visual awareness operationalized as the difference between event-related potentials (ERPs) in response to stimuli that enter awareness and stimuli that do not. There are three candidates for such a correlate: enhancement of P1 around 100 ms, enhancement of early posterior negativity around 200 ms (visual awareness negativity, VAN), and enhancement of late positivity (LP) in the P3 time window around 400 ms. Review of studies using different manipulations of awareness suggests that VAN is the correlate of visual awareness that most consistently emerges across different manipulations of visual awareness. VAN emerges also relatively independent of manipulations of nonspatial attention, but seems to be dependent on spatial attention. The results suggest that visual awareness emerges about 200 ms after the onset of visual stimulation as a consequence of the activation of posterior occipito-temporal and parietal networks. PMID- 20005251 TI - Hyperglycemia in diabetics and non-diabetics: effect on the risk for and severity of pneumococcal pneumonia. AB - OBJECTIVES: We sought to determine whether poor glucose control among diabetics is associated with increased risk for pneumococcal pneumonia and whether elevated admitting plasma glucose (APG) levels are associated with increased severity of this infection in diabetic and non-diabetic patients. METHODS: We compared hemoglobin A(1c) (HbA(1c)) in diabetics who had pneumococcal pneumonia with diabetic case-controls who did not have pneumonia. In patients with pneumococcal pneumonia, we related APG to disease severity as determined by SMART-COP score, need for ICU admission, and mortality at 7 and 30 days. RESULTS: Fifty-three of 233 patients with pneumococcal pneumonia (22.7%) were diabetic. Diabetics with pneumonia had poorer glycemic control than diabetic case-controls (HbA(1c) 8.2% vs. 7.2%, respectively, P<0.01). In pneumococcal pneumonia patients, SMART-COP scores, need for ICU admission, and mortality increased in proportion to the APG. These findings were attributable to the significant association between hyperglycemia and severity in non-diabetics. CONCLUSIONS: Poor glycemic control predisposes diabetics to pneumococcal pneumonia but, among diabetics, the degree of hyperglycemia at admission is not associated with increased disease severity. In contrast, among non-diabetics with pneumococcal pneumonia, hyperglycemia is a marker for severe disease and increased mortality, perhaps reflecting massive release of cytokines and glucocorticosteroids in overwhelming infection. PMID- 20005252 TI - Inter-individual differences in neurobiology as vulnerability factors for affective disorders: implications for psychopharmacology. AB - Susceptibility to affective disorders is individually different, and determined both by genetic variance and life events that cause significant differences in the CNS structure and function between individual subjects. Therefore it is plausible that search for the inter-individual differences in endophenotypes that mediate the effects of causal factors, both genetic and environmental, will reveal the substrates for vulnerability, help to clarify pathogenetic mechanisms, and possibly aid in developing strategies to discover better, more personalized treatments. This review first examines comparatively a number of animal models of human affect and affect-related disorders that rely on persistent inter individual differences, and then highlights some of the neurobiological findings in these models that are compatible with much of research in human behavioural and personality traits. Many behaviours occur in specific combinations in several models, but often remarkable dissociations are observed, providing a variety of constellations of traits. It is concluded that more systematic comparative experimentation on behaviour and neurobiology in different models is warranted to reveal possible "building blocks" of affect-related personality common in animals and humans. Looking into the perspectives in psychopharmacology the focus is placed on probable associations of inter-individual differences with brain structure and function, personality and coping strategies, and psychiatric vulnerability, highlighting some unexpected interactions between vulnerability endophenotypes, adverse life events, and behavioural traits. It is argued that further studies on inter-individual differences in affect and underlying neurobiology should include formal modeling of their epistatic, hierarchical and dynamic nature. PMID- 20005250 TI - Novel cell death signaling pathways in neurotoxicity models of dopaminergic degeneration: relevance to oxidative stress and neuroinflammation in Parkinson's disease. AB - Parkinson's disease (PD) is a common neurodegenerative movement disorder characterized by extensive degeneration of dopaminergic neurons in the nigrostriatal system. Neurochemical and neuropathological analyses clearly indicate that oxidative stress, mitochondrial dysfunction, neuroinflammation and impairment of the ubiquitin-proteasome system (UPS) are major mechanisms of dopaminergic degeneration. Evidence from experimental models and postmortem PD brain tissues demonstrates that apoptotic cell death is the common final pathway responsible for selective and irreversible loss of nigral dopaminergic neurons. Epidemiological studies imply both environmental neurotoxicants and genetic predisposition are risk factors for PD, though the cellular mechanisms underlying selective dopaminergic degeneration remain unclear. Recent progress in signal transduction research is beginning to unravel the complex mechanisms governing dopaminergic degeneration. During the 12th International Neurotoxicology meeting, discussion at one symposium focused on several key signaling pathways of dopaminergic degeneration. This review summarizes two novel signaling pathways of nigral dopaminergic degeneration that have been elucidated using neurotoxicity models of PD. Dr. Anumantha Kanthasamy described a cell death pathway involving the novel protein kinase C delta isoform (PKCdelta) in oxidative stress-induced apoptotic cell death in experimental models of PD. Dr. Ajay Rana presented his recent work on the role of mixed lineage kinase-3 (MLK3) in neuroinflammatory processes in neurotoxic cell death. Collectively, PKCdelta and MLK3 signaling pathways provide new understanding of neurodegenerative processes in PD, and further exploration of these pathways may translate into effective neuroprotective drugs for the treatment of PD. PMID- 20005253 TI - Heterogeneity in astrocyte morphology and physiology. AB - Astrocytes as a cell population are not well defined and comprise a heterogeneous population of cells. There are at least 9 different morphological variants which can coexist within one given brain region. Human astrocytes have a considerably more complex morphology as their rodent counterparts. There are also a number of functional differences depending on brain region and developmental stage in the normal (not pathologic) brain. Astrocytes can differ in functional gap junctional coupling, expression of transmitter receptors, membrane currents, and glutamate transporters. We feel that astrocyte heterogeneity has not yet been thoroughly explored and what we report here will just be a beginning of a new field of research. PMID- 20005255 TI - Technique for collection of cerebrospinal fluid from the cisterna magna in rat. AB - Cerebrospinal fluid (CSF) is in direct contact with the extracellular space in the central nervous system (CNS), and biological changes in the brain can be reflected in CSF. In the present article, a procedure for collection of CSF in rats is described. The technique quickly and reliably yields large quantities of CSF (50-150 microl) in rats. More importantly, blood contamination of the CSF is avoided. Furthermore, detections of ATP and interleukin (IL)-1 beta in the CSF have been carried out. ATP concentration in the CSF samples was between 8.3 and 15.8 nM, with an average of 10.5+/-0.83 nM (mean+/-SEM). The concentrations of IL 1beta were below the detection limit in the CSF in the laminectomy control rats, but it increased to 0.26+/-0.07 ng/ml at 1h after spinal cord injury. This technique offers an alternative method to surgical cannulation for the collection of CSF in rats. PMID- 20005256 TI - Modulation of immune response by combined targeting of complement receptors and low-affinity Fcgamma receptors. AB - Immune complexes (ICs) induce effective pathogen-specific innate and humoral immune response via immunecomplex-binding receptors, such as murine complement receptor type 1 and 2 (mCR1/2) and murine low-affinity Fc receptors for IgG (mFcgammaRII and III). The exact function of mCR1/2 in cooperation with mFcgammaRII/III in modulation of humoral immunity has not yet been adequately clarified. The aim of this study was to target these receptors by specific single chain fragments of antibody (scFv), either individually or in combination, thus modelling the action of IC. For targeting, we used scFv derived from the well characterized 7g6 and 2.4g2 monoclonal antibodies recognizing mCR1/2 and mFcgammaRII/III, respectively. These scFvs were monobiotinylated and conjugated to streptavidin or streptavidin-coated microspheres. Such complexes were investigated with respect to target receptor recognition and in vivo localization. Antibody response against the constructs was measured by ELISA and ELISPOT. Results show that targeting streptavidin complexes to mFcgammaRII/III induces stronger IgG1 response than targeting to mCR1/2 yet both strategies enhance the antibody response compared to the control group immunized with non targeted peptide-streptavidin complexes. Moreover, the immunogenicity of coupled antigens increased using microspheres as carrier, instead of using soluble streptavidin. In summery, our in vivo experiments reveal that mFcgammaRII/III is more potent a target than CR1/2 and show that combined targeting of CR1/2 and FcgammaRII/III receptors does not result in cumulative enhancement of the antigen specific immune response. In addition, microparticle-mediated enhancement of immunization can be further improved by FcgammaRII/III targeting. PMID- 20005254 TI - Effects of normal aging on prefrontal area 46 in the rhesus monkey. AB - This review is concerned with the effects of normal aging on the structure and function of prefrontal area 46 in the rhesus monkey (Macaca mulatta). Area 46 has complex connections with somatosensory, visual, visuomotor, motor, and limbic systems and a key role in cognition, which frequently declines with age. An important question is what alterations might account for this decline. We are nowhere near having a complete answer, but as will be shown in this review, it is now evident that there is no single underlying cause. There is no significant loss of cortical neurons and although there are a few senile plaques in rhesus monkey cortex, their frequency does not correlate with cognitive decline. However, as discussed in this review, the following do correlate with cognitive decline. Loss of white matter has been proposed to result in some disconnections between parts of the central nervous system and changes in the structure of myelin sheaths reduce conduction velocity and the timing in neuronal circuits. In addition, there are reductions in the inputs to cortical neurons, as shown by regression of dendritic trees, loss of dendritic spines and synapses, and alterations in transmitters and receptors. These factors contribute to alterations in the intrinsic and network physiological properties of cortical neurons. As more details emerge, it is to be hoped that effective interventions to retard cognitive decline can be proposed. PMID- 20005257 TI - The same well-characterized T cell epitope SIINFEKL expressed in the context of a cytoplasmic or secreted protein in BCG induces different CD8+ T cell responses. AB - Mycobacterium bovis BCG is still the most widely used vaccine against tuberculosis and CD8(+) T cells play important roles in fighting infection. We investigated how well antigen is processed and presented to CD8(+) T cells using the same well-characterized CD8(+) T cell epitope SIINFEKL expressed in either a cytoplasmic (GFP-OVA) or secreted (85B-OVA) context from BCG. We report that secreted SIINFEKL from 85B-OVA BCG is presented better than cytoplasmic SIINFEKL expressed by GFP-OVA BCG. PMID- 20005258 TI - Reduced expression of chemoattractant receptors by polymorphonuclear leukocytes in Hyper IgE Syndrome patients. AB - Hyper IgE Syndrome (HIES) is a rare genetic disorder, characterized by elevated serum IgE levels and reduced inflammatory responses to bacterial infections. This leads to dermatitis, recurrent lung infections and "cold abscesses". Recently, progress was made in HIES research, when mutations in STAT3 were found in the autosomal dominant form of HIES, and impaired responses of T helper 17 cells were reported. However, the causes for reduced inflammatory responses in these patients were not fully elucidated. In view of studies that indicated that polymorphonuclear leukocytes (PMN) of HIES patients are defective in their chemotactic properties, we asked if the PMN of these patients have reduced expression of receptors for chemoattractants. To analyze this possibility, we focused on fMLP and ELR(+)-CXC chemokines - which are essential for mounting acute inflammatory responses - and determined the coding sequences and expression levels of their corresponding receptors: FPR (for fMLP) as well as CXCR1 and CXCR2 (the receptors for ELR(+)-CXC chemokines). The analyses of these receptors in HIES patients indicated that their coding sequences were intact and normal. However, the percentages of PMN that expressed FPR, CXCR1 and CXCR2 were significantly lower in HIES patients. In addition, lower expression levels per cell were denoted for CXCR1 in PMN of the patients. A cumulative score that was calculated for the three chemoattractant receptors together indicated that in some of the patients there were prominent reductions, of up to approximately 50% in the overall expression of the receptors (indicated by % positive cells and mean expression levels per cell). In addition, we asked whether deregulation of PMN activities in HIES may result from binding of IgE to corresponding receptors on HIES PMN. Our findings indicate that this is probably not the case, because similarly to normal PMN, the cells of HIES patients did not express notable levels of the IgE receptors FcvarepsilonRI and FcvarepsilonRII. Together, these results provide novel information on the expression of key determinants in PMN migration in HIES, suggesting that a defect in the expression of chemoattractant receptors may lead to impaired chemotaxis found in HIES patients, and to decreased inflammatory responses. PMID- 20005259 TI - Tribbles-1 and -2 are tumour suppressors, down-regulated in human acute myeloid leukaemia. AB - Constitutive MAPK signalling is observed in approximately 50% of acute myeloid leukaemia (AML) cases. JNK activation in particular is associated with treatment failure in AML. Tribbles proteins (trb-1, trb-2 and trb-3) are potent negative regulators of MAPK pathways influencing apoptosis, differentiation and cell-cycle progression. Here we aimed to examine tribbles gene expression in AML and to characterise their role in leukaemic cells. A microarray dataset was interrogated for tribbles expression levels in AML cases and healthy controls. Myeloid cell proliferation and apoptosis were assayed in response to trb-1/trb-2 gene knockdown and overexpression, as well as a physical and functional interaction between trb and C/EBPalpha. Trb-2 expression was reduced in AML compared to healthy controls (correlating with nucleophosmin (NPM1) mutations), while low trb 1 expression was associated with inactive C/EBPalpha. In vitro assays indicated that trb-1/trb-2 are growth restrictive and pro-apoptotic in Me-1 cells, each capable of inhibiting JNK activation. JNK inactivation was itself associated with reduced Bcl-2 Ser70 phosphorylation, a residue which, when phosphorylated, maintains the anti-apoptotic activity of Bcl-2. Consistent with this, tribbles mediated dephosphorylation of Bcl-2 Ser70 was associated with subsequent apoptosis. Trb-1/trb-2 transcription appeared to be moderately C/EBPalpha responsive, and physical interaction between C/EBPalpha and trb-1/trb-2 was observed, suggesting a potential for auto-regulation of trb-1 and trb-2 transcription. In conclusion, we propose that trb-1 and trb-2 tumour suppressor activity may be abrogated in a proportion of AML patients. This may lead to enhanced cell survival, and therefore contribute to pathogenesis of the disease. Trb-1/trb-2 may, therefore, represent useful therapeutic targets for the treatment of AML in patients with dys-regulated trb activity. PMID- 20005260 TI - Characterization of the mixture of genotypes of a Citrus tristeza virus isolate by reverse transcription-quantitative real-time PCR. AB - A multiplex real-time PCR assay was developed to detect and quantify the Citrus tristeza virus (CTV) genotypic mixture present in infected plants. CTV isolate FS627, a complex Florida isolate containing T36, T30 and VT genotypes and its aphid transmitted subisolates was used. The relative quantitative assay was carried out using specific primers and probes developed from the genotypes of three CTV virus isolates and included the coat protein region of isolate T36 and the 5' end, ORF 1a and ORF 2 region of isolates T36, T30 and VT. Among the three genotypes present in the aphid transmitted subisolates, the T30 genotype showed higher overall relative quantitation in all specific regions compared to other isolates. The profiles of the some aphid transmitted subisolates were different from the parent source from which they transmitted. The 2(-DeltaDeltaCt) method (the amount of target, normalized to an endogenous control and relative to a calibrator) was used to analyze the relative titers of the three reference genotypes in the aphid transmitted plants infected with FS627. This protocol enabled assessments of CTV genetic diversity in the aphid transmitted subisolates. This simple quantitative assay was sensitive, efficient, and took less time than other existing methods. This relative quantitative assay will be a reliable tool for diagnosis, detection and genetic diversity studies on CTV. PMID- 20005261 TI - Pancreatic beta-cell responses to GLP-1 after near-normalization of blood glucose in patients with type 2 diabetes. AB - This study investigated the effects of strict glycaemic control on beta-cell function in nine obese subjects with type 2 diabetes (T2DM), using graded glucose infusions together with infusions of saline or GLP-1 before (HbA(1)c: 8.0+/-0.4%) and after four weeks of near-normalization of blood glucose (BG) using insulin (mean diurnal BG: 6.4+/-0.3 mmol/l; HbA(1)c: 6.6+/-0.3%). Nine matched healthy subjects acted as controls. In controls, area-under-curve (AUC) for amylin, C peptide and proinsulin were higher with GLP-1 than saline (P<0.001). The AUC amylin/C-peptide ratio was similar on both days, while AUC proinsulin/C-peptide ratio was higher with GLP-1 (P=0.02). In the patients, amylin, C-peptide and proinsulin AUCs were unaltered by near-normoglycaemia per se. Proinsulin responses to GLP-1 were unchanged, but amylin and C-peptide AUCs increased (P<0.05) after insulin treatment, and AUC amylin/C-peptide ratios rose to control levels. Near-normoglycaemia tended to reduce AUC proinsulin/C-peptide ratio, which was significant (P=0.04) with GLP-1, but still higher than with saline (P=0.004). In conclusion, amylin, C-peptide and proinsulin responses to glucose were unaffected by four weeks of near-normoglycaemia, whereas GLP-1 increased amylin and C-peptide secretion and amylin/C-peptide ratio. Near-normoglycaemia reduced proinsulin/C-peptide ratio during stimulation with GLP-1, suggesting that strict glycaemic control might ameliorate some of the disturbances in beta-cell function characterizing T2DM. PMID- 20005262 TI - Liraglutide, but not vildagliptin, restores normoglycaemia and insulin content in the animal model of type 2 diabetes, Psammomys obesus. AB - In order to investigate the effect and mechanism of liraglutide and vildagliptin in diabetic Psammomys obesus, we examined proliferation and apoptosis of beta cells, beta-cell mass (BCM), and pancreatic insulin content after zero, six and fourteen days of treatment compared to control groups. One group of animals was kept on low-energy diet and seven groups were given high-energy diet (HED) that induced diabetes over a four week period. Non-fasting morning blood glucose, body weight, HbA(1C) and pancreatic insulin content were measured and beta cell mass (BCM), proliferation and apoptosis frequencies were determined using stereological point counting. Liraglutide significantly reduced blood glucose and even normalized it in all animals treated for six days and in 11 out of 17 animals treated for fourteen days. HED increased BCM and treatment with liraglutide did not change this. However, compared to the vehicle-treated animals pancreatic insulin content was normalized in animals treated for six and fourteen days with liraglutide. In contrast, vildagliptin, in doses causing full inhibition of plasma DPP-IV activity, neither reduced blood glucose nor altered HED-induced increases in BCM or pancreatic insulin content. These results suggest that liraglutide restores normoglycaemia and improves glycaemic control in P. obesus by increasing their insulin content and improving the function of the beta cells. In contrast, vildagliptin does not improve glycaemic control in P. obesus nor affect beta-cell insulin content. PMID- 20005263 TI - Ubiquitin-proteasome-mediated degradation and synthesis of MyoD is modulated by alphaB-crystallin, a small heat shock protein, during muscle differentiation. AB - alphaB-crystallin, a small heat shock protein, plays an important role in muscle homeostasis. It gets up-regulated during muscle differentiation and mice lacking alphaB-crystallin die prematurely with extensive muscle wastage. We have examined the role of alphaB-crystallin in muscle development using C2C12 myoblasts as a model system. Over-expression of alphaB-crystallin delays the muscle differentiation program significantly. C2C12 myoblasts over-expressing alphaB crystallin (CRYAB-C2C12) display defect in cell-cycle exit upon induction of differentiation. During differentiation, CRYAB-C2C12 cells exhibit sustained level of cyclin D1 and delay in p21 and myogenin expression as compared to C2C12 cells. We find less accumulation of MyoD in CRYAB-C2C12 cells than in C2C12 cells. In vivo protein stability studies reveal faster ubiquitin-proteasome mediated MyoD degradation in CRYAB-C2C12 cells (t(1/2)=1.42 h) than in C2C12 cells (t(1/2)=2.37 h). Immuno-precipitation experiments showed that MyoD gets ubiquitinated at earlier time points in CRYAB-C2C12 cells than in C2C12 cells. Our data reveal alterations in the synthesis and degradation of MyoD in CRYAB C2C12 cells. The level of alphaB-crystallin as well as its Ser-59 phosphorylated form increases with increasing time of differentiation. Our studies show, inter alia, that alphaB-crystallin modulates myogenesis by altering MyoD level and provide an interesting insight in its role in myogenesis. PMID- 20005264 TI - A curve fitting method of quantifying green fluorescent protein expression level in Escherichia coli. AB - Green fluorescent protein (GFP) has all the essential properties of a quantitative reporter protein and the fluorescence of GFP is a reliable and quantitative reporter of underlying differences in expression levels. However, GFP fluorescence does not increase with cell density in direct proportion because of its fluorescence quenching. And the fluorescence quenching is always ignored by most GFP fluorescence assays that provide a measure of average fluorescence intensity over an entire sample cell population. We now propose a novel method that accurately quantifies the fluorescence intensity of GFP expressed in Escherichia coli by a fluorescence spectrophotometer. In our method, a cell containing GFP was regarded as a fluorochrome and the data processing was essentially different from the previous methods. The experimental assay data were curve fitted to calibrate the mean fluorescence intensity of a cell population. Thus, the impact of fluorescence quenching caused by cell density was amended. Moreover, the mean fluorescence intensity of a cell population was roughly proportional to the fluorescence quantum efficiency, a property of a fluorochrome, so it can be used to evaluate the GFP expression level in a given cell population. PMID- 20005265 TI - A novel brain heart infusion broth supports the study of common Francisella tularensis serotypes. AB - Francisella tularensis Schu S4, LVS and U112 have become model organisms for the study of Francisella pathogenesis, and represent a cross section of the different F. tularensis subspecies. Both Schu S4 and LVS are fastidious organisms, requiring medium fortified with supplements and nutrients for enhanced growth. Chamberlains defined medium, Tryptone Soy Broth supplemented with cysteine (TSBc), and cation-adjusted Mueller-Hinton broth (CAMHB) supplemented with 2% IsoVitaleX are typically used in the cultivation of these bacteria. In this report, we describe a simple brain heart infusion broth formulation that can be used to obtain superior growth characteristics in all of these model organisms, and can support bacterial growth from low inoculum. Surprisingly, CAMHB, which is favored in the literature for culturing Schu S4 and LVS, induced the worst growth characteristics of the four formulations studied. To expand on these observations, an additional seven strains of F. tularensis, representing types A.I, A.II, and B were selected from the Department of Defense United Culture Collection (UCC) and a comparative analysis of their growth characteristics performed in the four broth formulations. Results demonstrate differences in the growth characteristics of Francisella species that are significantly influenced by both strain type and the choice of growth medium. Though four of the five additional Type A strains displayed superior growth characteristics in Chamberlain's defined medium, growth characteristics of all three model organisms, as well the Type B strains, were enhanced by the new BHI-based broth formulation. We conclude that this medium represents the optimal choice for cultivation of the three model organisms used for Francisella research. PMID- 20005266 TI - In vitro detection and primary cultivation of bacteria producing materials inhibitory to ruminal methanogens. AB - A novel method for screening bacterial isolates for their potential to inhibit the growth of ruminal methanogenic Archaea was developed using a modification of the soft agar overlay technique, formally used for the isolation of lytic bacteriophages. This method may be used in the specific, hydrogen-rich conditions required for the growth of ruminal methanogenic Archaea. PMID- 20005267 TI - Sustained gamma-band EEG following negative words in depression and schizophrenia. AB - INTRODUCTION: Sustained and elaborative emotional information processing in depression and decreased affective elaboration in schizophrenia are considered hallmarks of these disorders but have not been directly measured. Gamma-band (35 45 Hz) EEG has been associated with semantic functions such as feature binding and may index these elaborative processing. This study examined whether there were group differences in baseline and sustained gamma-band EEG following emotional stimuli in healthy adults as well as adults with depression and schizophrenia. METHODS: 24 never-depressed healthy controls, 14 patients with DSM IV unipolar major depressive disorder, and 15 patients with DSM-IV schizophrenia completed a lexical emotion identification task during EEG assessment. Gamma-band (35-45 Hz) EEG in response to negative words was the primary dependent measure. RESULTS: As predicted, depressed individuals displayed sustained and increased gamma-band EEG throughout the task, and particularly in the seconds following negative words. Individuals with schizophrenia displayed decreased gamma-band activity throughout the task. CONCLUSIONS: These data suggest that gamma-band EEG, measured over several seconds, may serve as a useful index of sustained semantic information processing. Depressed individuals appear to engage in sustained elaboration following emotional stimuli, whereas individuals with schizophrenia are not as prone to this type of elaborative processing. PMID- 20005268 TI - Constantine A. Mangina, President, I.O.P./UN (New York). PMID- 20005269 TI - Luciferin liposomes for enhanced in vivo bioluminescence. PMID- 20005270 TI - Grafting of poly(ethylene glycol) to poly-lysine augments its lifetime in blood circulation and accumulation in tumors without loss of the ability to associate with siRNA. AB - Poly-lysine has been studied as a carrier for the delivery of drugs and nucleic acids for at least a decade. It is an especially attractive carrier for DNA and RNA, because of its condensed cationic charges. In our previous study, we showed that poly(ethylene glycol) (PEG) grafted to poly-L-lysine (PLL) remarkably increased the life time of a small interfering RNA (siRNA) in blood circulation. In this study, we prepared a new series of PEG-grafted PLL (PLL-g-PEG) with various lengths (PEG 2kDa, 5kDa, and 10kDa and PLL 28kDa and 40kDa), to evaluate masking effects of PEG on cationic charges of PLL in vivo and the structural implications for biodistribution and tumoral accumulation. The best in the series, 40K10P37 (40kDa of PLL, 10kDa of PEG, 37mol% grafting) with molecular weight of 10(6) as determined by Multi-Angle Laser Light Scattering (MALLS), accumulated in tumors at about 8% of the injected dose per gram of tissue. Interestingly, a PLL-g-PEG conjugate pre-mixed with murine sera prevented degradation of siRNA, suggesting that PLL-g-PEG preferentially associates with siRNA in sera. Our results indicate grafting of PEG to the side chains of PLL augments its lifetime in blood circulation and tumoral accumulation without loss of the ability to associate with siRNA and support further evaluation of these cationic delivery carriers. PMID- 20005271 TI - Do HPMA copolymer conjugates have a future as clinically useful nanomedicines? A critical overview of current status and future opportunities. AB - N-(2-Hydroxypropyl)methacrylamide (HPMA) copolymer conjugates containing doxorubicin designed in the late 1970s/early 1980s as anticancer polymer therapeutics were the first synthetic polymer-based anticancer conjugates to enter clinical trial beginning in 1994. Early clinical results were promising, confirming activity in chemotherapy refractory patients and the safety of HPMA copolymers as a new polymer platform in this setting. Subsequent Phase I/II trials have investigated conjugates containing paclitaxel (PNU 166945), camptothecin (PNU 166148) (both failed in clinical trials underlining the importance of rational design), and most recently HPMA-copolymer platinates (AP5280 and then AP5346-ProLindac(TM)) entered Phase II clinical development. There are a growing array of second generation HPMA copolymer-based systems involving combination therapy, incorporating putative targeting ligands, having an ever more complex architecture, and both drug and protein conjugates are being proposed as novel treatments for diseases other than cancer. Despite their promise, and the success of polymeric drugs and polymer-protein conjugates, no polymer-drug conjugate (HPMA copolymer-based or otherwise) has yet entered routine clinical use. It is timely to reflect on the progress made over the last 30 years, the relative merits of HPMA copolymers as a platform compared to other polymeric carriers, and comment on their future potential as polymer-based nanomedicines into the 21st century in comparison with the many alternative strategies now emerging for creation of nanopharmaceuticals. PMID- 20005272 TI - HPMA copolymers for modulating cellular signaling and overcoming multidrug resistance. AB - Unique properties of N-(2-hydroxypropyl)methacrylamide (HPMA) copolymer granted its wide use as a carrier for different nanotherapeutics. However, the role of HPMA in a drug delivery system is not limited solely to a carrier for an active payload. Detailed investigations revealed its deep influence on the molecular mechanisms of antitumor action of chemotherapeutic drugs bound to HPMA copolymer. Such influence involves changing the internalization and intracellular trafficking of an entire HPMA-drug complex, modifying the topography of the accumulation of delivered anticancer drug in the tumor and inside tumor cells, overcoming and suppression of existing drug resistance and preventing its de novo development, inhibition of cellular drug detoxification and intracellular repair mechanisms, affecting cell death signaling pathways and mechanisms of apoptosis and necrosis induction. The present review underlines the major mechanisms of the aforementioned effects leading to the substantial enhancement of cell death inducing ability of conjugated anticancer drugs. PMID- 20005273 TI - HPMA copolymers: 30 years of advances. PMID- 20005274 TI - Components of attentional bias for food cues among restrained eaters. AB - The study aimed to investigate attentional bias for food cues among restrained eaters. In particular, the roles of speeded detection (enhanced orientation of attention toward food stimuli) and slowed disengagement (trouble disengaging attention from food stimuli) were examined. Participants were 78 female undergraduate students aged 18-25 years, classified as restrained (N=38) or unrestrained eaters (N=40). Attentional bias was assessed by a visual search task which required participants to locate the position of an odd-one-out target word in a matrix of 19 distractor words. Restrained eaters were disproportionately faster than unrestrained eaters to detect a food word within a neutral matrix compared to a neutral word within a neutral distractor matrix. Restrained eaters were also disproportionately faster, rather than slower, than unrestrained eaters to detect a neutral word within a food matrix compared to a neutral word within a neutral distractor matrix. Thus restrained eaters show a heightened vigilance for food cues, but no slower disengagement from such cues. PMID- 20005275 TI - Emotional Appetite Questionnaire. Construct validity and relationship with BMI. AB - The Emotional Appetite Questionnaire (EMAQ) comprises ratings of tendency to eat in response to both positive and negative, emotions and situations. To assess construct validity, the responses of 232 male and female participants to the EMAQ subscales were correlated with the subscales of the Dutch Eating Behavior Questionnaire (DEBQ), which has been extensively validated. In addition, the EMAQ scores were correlated with BMI. Convergent validity was demonstrated by a significant positive correlation between the negative emotions and situations scores of the EMAQ and the emotional eating subscale score of the DEBQ (DEBQ-E). Moreover, discriminant validity was demonstrated by low correlations of EMAQ positive emotions and situations scores with the DEBQ-E score. For the study sample, the EMAQ negative scores were significantly positively correlated with BMI, and the EMAQ positive scores were significantly inversely correlated with BMI. As BMI increased so did reported negative emotional and situational eating whereas as BMI decreased, reported positive emotional and situational eating increased. Although causality cannot be inferred from correlations, eating more under negative emotions may contribute to being overweight whereas eating less may contribute to being underweight. The EMAQ was shown to have construct validity, and emotional eating was significantly correlated with BMI. PMID- 20005276 TI - Inaccessible food cues affect stress and weight gain in calorically-restricted and ad lib fed rats. AB - Research suggests that caloric restriction (CR) is beneficial; however, the effects of CR in the context of food cues are unclear. A 2 (food cue vs. no cue)x2 (CR vs. ad lib) between-subjects design was employed to test these effects in 40 rats. It was predicted that cue exposure and CR would induce stress, and that these factors might interact synergistically. The results demonstrated that cue-exposed CR rats weighed less than did non-exposed CR rats. A blunted stress response was evident in CR rats relative to ad lib rats. Finally, cue-exposed rats had higher corticosterone levels and body weight during ad lib feeding than did non-cued rats. These results suggest that both CR and chronic food-cue exposure can be stressful, and the implications of this research are discussed in the context of humans' 'obesigenic' environment. PMID- 20005277 TI - Excessive appetitive arousal in Prader-Willi syndrome. AB - This study focused on genetic and behavioural aspects of one important component of the motivation to eat - how appetitive arousal is elicited through the presentation of food-associated stimuli. Individuals with Prader-Willi syndrome, a genetic disorder associated with hyperphagia, and control participants completed a computerised response task in the presence of motivational stimuli. In controls, appetitive arousal was specific to particular stimuli. In contrast, individuals with PWS showed a non-specific pattern of arousal. Over-activation of the anticipatory motivation system may be one consequence of the genetic disorder in PWS. PMID- 20005278 TI - Pediatric neurological syndromes and inborn errors of purine metabolism. AB - This review is devised to gather the presently known inborn errors of purine metabolism that manifest neurological pediatric syndromes. The aim is to draw a comprehensive picture of these rare diseases, characterized by unexpected and often devastating neurological symptoms. Although investigated for many years, most purine metabolism disorders associated to psychomotor dysfunctions still hide the molecular link between the metabolic derangement and the neurological manifestations. This basically indicates that many of the actual functions of nucleosides and nucleotides in the development and function of several organs, in particular central nervous system, are still unknown. Both superactivity and deficiency of phosphoribosylpyrophosphate synthetase cause hereditary disorders characterized, in most cases, by neurological impairments. The deficiency of adenylosuccinate lyase and 5-amino-4-imidazolecarboxamide ribotide transformylase/IMP cyclohydrolase, both belonging to the de novo purine synthesis pathway, is also associated to severe neurological manifestations. Among catabolic enzymes, hyperactivity of ectosolic 5'-nucleotidase, as well as deficiency of purine nucleoside phosphorylase and adenosine deaminase also lead to syndromes affecting the central nervous system. The most severe pathologies are associated to the deficiency of the salvage pathway enzymes hypoxanthine guanine phosphoribosyltransferase and deoxyguanosine kinase: the former due to an unexplained adverse effect exerted on the development and/or differentiation of dopaminergic neurons, the latter due to a clear impairment of mitochondrial functions. The assessment of hypo- or hyperuricemic conditions is suggestive of purine enzyme dysfunctions, but most disorders of purine metabolism may escape the clinical investigation because they are not associated to these metabolic derangements. This review may represent a starting point stimulating both scientists and physicians involved in the study of neurological dysfunctions caused by inborn errors of purine metabolism with the aim to find novel therapeutical approaches. PMID- 20005279 TI - The loudness dependence of the auditory evoked potential (LDAEP) in schizophrenia, bipolar disorder, major depressive disorder, anxiety disorder, and healthy controls. AB - BACKGROUND: Serotonergic dysfunction in schizophrenia, bipolar disorder, major depressive disorder, anxiety disorder, and healthy controls was evaluated by measuring the activity of the loudness dependence of the auditory evoked potential (LDAEP). METHODS: The 357 subjects who were evaluated comprised 55 normal controls, 123 patients with major depressive disorder, 37 with bipolar disorder, 46 with schizophrenia, 37 with panic disorder (PD), 31 with generalized anxiety disorder (GAD), and 28 with post-traumatic stress disorder (PTSD). RESULTS: LDAEP was significantly stronger in healthy controls than in patients with either bipolar disorder (p=0.025) or schizophrenia (p=0.008), and significantly stronger in patients with major depressive disorder than in those with bipolar disorder (p=0.01) or schizophrenia (p=0.03). LDAEP did not differ significantly between patients with major depressive disorder and healthy control subjects (p=0.667), or between healthy control subjects and patients with anxiety disorder, including PD (p=0.469), GAD (p=0.664), and PTSD (p=0.167). CONCLUSION: The findings of the present study reveal that patients with major psychiatric disorders exhibit different strengths of LDAEP according to their serotonin related pathology. Studies controlled for psychotropic medication, menstruation cycle, and smoking are needed. PMID- 20005280 TI - Adding a low dose atypical antipsychotic drug to an antidepressant induced a rapid increase of plasma brain-derived neurotrophic factor levels in patients with treatment-resistant depression. AB - Only two-thirds of depressive patients respond to antidepressant treatment. Recently, addition of an atypical antipsychotic drug to ongoing treatment with an antidepressant has been considered effective and well-tolerated. In the present study, we examined the effects of various atypical antipsychotic drugs as adjuvant to antidepressants, including selective serotonin reuptake inhibitors (SSRIs), serotonin noradrenaline reuptake inhibitors, tricyclic antidepressants and mood stabilizers, on plasma BDNF levels in refractory depressed patients. Forty-five patients who met the DSM-IV criteria for major depressive disorder (n=31) or bipolar disorder (10 with bipolar I, 4 with bipolar II) were enrolled in the study. Twenty-one were male and 24 were female, and their ages ranged from 28 to 71 (mean+/-SD=49+/-12) years. Plasma BDNF levels were measured using a sandwich ELISA. The plasma BDNF levels in responders (those showing a decline in HAM-D scores of 50% or more) were significantly increased 4weeks after the administration of each atypical antipsychotic drug, while the levels in non responders were not changed. Furthermore, there was a significant correlation between the changes in HAM-D scores and the changes in plasma BDNF levels. These results suggest that adding an atypical antipsychotic drug to ongoing treatment with an antidepressant or mood stabilizer is useful and well-tolerated for refractory depressed patients, and the efficacy of atypical antipsychotics as an adjuvant might involve an increase of plasma BDNF levels. PMID- 20005281 TI - Intermittent dosing of G-CSF to ameliorate carbon tetrachloride-induced liver fibrosis in mice. AB - On the basis of the recent report that granulocyte colony-stimulating factor (G CSF) administration after rats' partial orthotopic liver transplantation greatly improved survival rate and liver regeneration of partial graft, we here evaluated the effect of intermittent administration of G-CSF on fibrosis formation induced by carbon tetrachloride (CCl(4)). Bone marrow chimeric female C57BL/6 mice were treated with G-CSF at days 1, 7, 14, 21, and 28 after CCl(4) challenge. At day 35 after CCl(4) administration, we found that G-CSF treatment significantly reduced CCl(4)-induced liver damage and collagen deposition. In addition, levels of hepatic hydroxyproline and serum fibrosis markers in mice receiving G-CSF administration after CCl(4) challenge were significantly lower compared to those of control mice. Histological examination suggested that hepatic damage recovery was much better in these G-CSF-treated mice. Immunofluorescence and fluorescence in situ hybridization (FISH) analysis revealed that donor cells engrafted into host liver, had epithelium-like morphology and expressed albumin, although at low frequency. These results suggest that intermittent G-CSF treatment might initiate endogenous hepatic tissue regeneration in response to CCl(4) injury and ameliorate its fibrogenic effects. PMID- 20005282 TI - Telomerase and reactive oxygen species: Comments on Saretzki, G., 2009. Telomerase, mitochondria and oxidative stress. Exp. Gerontol. 44, 485-492. PMID- 20005283 TI - Acceleration of brain amyloidosis in an Alzheimer's disease mouse model by a folate, vitamin B6 and B12-deficient diet. AB - Epidemiological and clinical studies indicate that elevated circulating level of homocysteine (Hcy) is a risk factor for developing Alzheimer's disease (AD). Dietary deficiency of folate, vitamin B6 and B12 results in a significant increase of Hcy levels, a condition also known as hyperhomocysteinemia (HHcy). In the present study we tested the hypothesis that a diet deficient for these three important factors when administered to a mouse model of AD, i.e. Tg2576, will result in HHcy and in an acceleration of their amylodotic phenotype. Compared with Tg2576 mice on regular chow, the ones receiving the diet deficient for folate, B6 and B12 developed HHcy. This condition was associated with a significant increase in Abeta levels in the cortex and hippocampus, and an elevation of Abeta deposits in the same regions. No significant changes were observed for steady-state levels of total APP, BACE-1, ADAM-10, PS1 and nicastrin in the brains of mice with HHcy. No differences were observed for the main Abeta catabolic pathways, i.e. IDE and neprilysin proteins, or the Abeta chaperone apolipoprotein E. Our findings demonstrate that a dietary condition which leads to HHcy may also result in increased Abeta levels and deposition in a transgenic mouse model of AD-like amylodosis. They further support the concept that dietary factors can contribute to the development of AD neuropathology. PMID- 20005285 TI - Long-term inhibition of dipeptidyl peptidase-4 in Alzheimer's prone mice. AB - We tested here the impact of a long-term inhibition of dipeptidyl peptidase-4 (DPP-4) with sitagliptin on the deposition of amyloid-beta within the brain and deficits in memory-related behavioral paradigms in a model of Alzheimer's disease (AD): double transgenic mice B6*Cg-Tg(APPswe,PSEN1dE9)85Dbo/J. Mice began to receive sitagliptin at 7 months of age. Three different dose of sitagliptin (5, 10 and 20 mg/kg), were administered daily for 12 weeks by gastric gavage. The treatments counteracted: (i) the memory impairment in the contextual fear conditioning test; (ii) increased the brain levels of GLP-1; (iii) produced significant reductions of nitrosative stress and inflammation hallmarks within the brain, as well as (iv) a significant diminution in the ultimate number and total area of betaAPP and Abeta deposits. All these effects much more evident for the dose of 20 mg/kg sitagliptin. The long-term inhibition of the endogenous DPP 4 enzymes with sitagliptin can significantly delay some forms of AD pathology, including amyloid deposition, when administered early in the disease course of a transgenic mouse model of AD. PMID- 20005284 TI - Effect of caloric restriction on base-excision repair (BER) in the aging rat brain. AB - Apyrimidinic/apurinic endonuclease (APE) is a key protein involved in the base excision DNA repair (BER) pathway of oxidative DNA lesions. Using a novel oligonucleotide substrate, we demonstrate that APE activity in the frontal/parietal cortex (F/PCTX), cerebellum, brainstem, midbrain and hypothalamus declined with age in rats on an ad libitum (AL) diet. In contrast, APE activity for these brain regions was approximately 1.5-3 times higher in young, caloric restricted (CR) rats. Despite continuous CR treatment in all animals since six weeks of age, APE activity in the CR group started to decline by middle-age and continued into old age. However, CR maintained APE activity at a level that was significantly higher than that in AL rats across age and in the brain regions examined. Because Western analysis of APE, DNA polymerase beta and DNA ligase III levels in the F/PCTX of both CR and AL rats remained unchanged with age, this suggests that the increased APE activity in CR rats is the result of differential post-translational modification of APE. PMID- 20005286 TI - Substrate adaptation of Trichophyton rubrum secreted endoproteases. AB - Trichophyton rubrum is the most common pathogen caused the dermatophytosis of nail and skin in human. The secreted proteases were considered to be the most important virulence factors. However, the substrates adaptation of T. rubrum secreted proteases is largely unknown. For the first time, we use the keratins from human nail and skin stratum corneum as the growth medium to investigate the different expression patterns of T. rubrum secreted endoproteases genes. During grow in both keratin-containing media SUB7 and MEP2 were the highest expressed gene in each family. These results indicated that SUB7 and MEP2 may be the dominant endoproteases secreted by T. rubrum during host infection and the other proteases may play a supplementary role. The direct comparison of T. rubrum grown on skin and nail medium showed different substrate favorite of secreted endoproteases. The genes MEP2, SUB5, SUB2 and SUB3 were more active during growth in skin medium, possibly these proteases have a higher affinity for skin original keratins. While the structures of SUB1, SUB4, and MEP4 maybe more suitable for the degradation of nail original keratins. This work presents useful molecular details for further understanding the pathogenesis of secreted proteases and the wide adaptation of T. rubrum. PMID- 20005287 TI - Serum cytokine profile in hepatitis C virus carriers presenting cryoglobulinaemia and non-organ-specific autoantibodies. AB - This work investigated the serum cytokine profile (IL-2, IL-4, IL-5, IL-10, IFN gamma and BAFF) of hepatitis C virus (HCV) carriers with autoimmunity. Forty seven HCV carriers and 28 healthy controls were evaluated. Cytokine levels were measured by ELISA. Patients and controls presented similar levels of IL-2, IL-4, IL-5, IL-10, IFN-gamma and BAFF (p>0.05). Cryoglobulinaemic HCV carriers had increased IL-2 (p=0.013), IL-5 (p=0.018) and BAFF (p=0.050). IFN-gamma level was decreased in HCV carriers with rheumatoid factor in comparison with those that were RF-seronegative (p=0.035). Patients with beta2GPI IgA antibodies when were compared with those without this autoantibody, had more serum IL-2 (p=0.009), IL 5 (p=0.018) and BAFF (p=0.039). Interleukin-2 was increased in HCV carriers with positive ANA when they were compared with ANA-seronegative carriers (p=0.044). Interleukins IL-4 and IL-10 were not associated with autoimmunity (P>0.05). In HCV carriers, IL-2 was correlated with IL-5 (p<0.0001) and IFN-gamma (p=0.015), and IL-5 with IFN-gamma (p=0.015). We concluded that the serum profile of cytokines in HCV carriers presenting autoimmune markers may be mainly represented by increased IL-2, IL-5 and BAFF. PMID- 20005288 TI - Cartilage tissue engineering for auricular reconstruction: in vitro evaluation of potential genotoxic and cytotoxic effects of scaffold materials. AB - Tissue engineering of autologous cartilage transplants is suggested as a new approach in reconstruction of external auricular deformities. 1.6-Hexanediol (HD), 1.8-diazabicyclo[5.4.0]undec-7-ene (DBU) and 6-hydroxyhexanoic acid (HHA) are matrices of the open-pored polyurethane three-dimensional scaffold. Since these bioresorbable materials may interact with the human organism, cytotoxic effects on human chondrocytes and lymphocytes and genotoxic effects on human lymphocytes were monitored. Staining with propidium iodide and fluorescence diacetate as well as the EZ4U proliferation assay served for the detection of cytotoxic effects of the materials on human chondrocytes. Trypan blue staining was used to monitor cytotoxicity on lymphocytes. Genotoxic effects on lymphocytes in terms of strand breaks, alkali labile sites and incomplete excision repair were determined by the alkaline single cell microgel electrophoresis (Comet) assay. Cytotoxic effects in chondrocytes and lymphocytes as well as genotoxic effects in lymphocytes were dose-dependent with threshold values of 5mg/mL HD, 0.5mg/mL DBU and 0.03 mg/mL HHA showing no effects. These data suggest that these matrices could be safely used for scaffolds made of polyurethane unless these compounds are not released at a rate giving higher concentrations at the site of implantation or in body fluids, respectively. PMID- 20005289 TI - Evodiamine-induced human melanoma A375-S2 cell death was mediated by PI3K/Akt/caspase and Fas-L/NF-kappaB signaling pathways and augmented by ubiquitin-proteasome inhibition. AB - Evodiamine, a major alkaloidal component of Evodiae fructus exhibits anti-tumor activities. We have previously reported that evodiamine has a marked inhibitory effect on IL-1 sensitive human melanoma A375-S2 cells proliferation, and this action might be through inactivation of PI3K signaling. However, the detailed molecular mechanisms of evodiamine-induced cell death remains poorly understood. In present study, we further confirmed that Akt is the main effector molecule involved in this pathway. Evodiamine also led to IkappaBalpha phosphorylation and degradation that reflect translocation of NF-kappaB. Pretreatment of A375-S2 cells with ubiquitin-proteasome inhibitor MG132 was shown to aggregate the evodiamine caused cell death at 24h. In addition, MG132 reduced ERK phosphorylation, increased caspase-3 activation, Fas-L expression and Bcl-2 cleavage in evodiamine-treated A375-S2 cells. These results suggested the PI3K/Akt/caspase and Fas-L/NF-kappaB signaling pathways might account for the responses of A375-S2 cell death induced by evodiamine, and these signals could be augmented by ubiquitin-proteasome pathway. PMID- 20005290 TI - Genotoxicity of urethane dimethacrylate, a tooth restoration component. AB - Urethane dimethacrylate (UDMA) is used in dental restorative materials in its polymeric form. However, the process of polymerization is usually incomplete and the monomers of UDMA can diffuse into the oral cavity and the pulp, reaching millimolar concentrations. In the present work we showed that UDMA at 0.1 and 1.0 mM decreased the viability of and induced DNA damage in lymphocytes in a concentration dependent manner, but it did not affect a plasmid DNA in vitro. UDMA at 1mM induced apoptosis in lymphocytes. The lymphocytes exposed to UDMA were able to repair their DNA within 60 min. Analysis with DNA repair enzymes Endo III and Fpg showed that UDMA induced mainly oxidative DNA lesions. Vitamin C and chitosan decreased genotoxic effect of UDMA. Our results show that monomers of UDMA may exert pronounced cyto- and genotoxic effects in human lymphocytes and chitosan can be considered as a protection against such effects. PMID- 20005291 TI - Nigella sativa thymoquinone-rich fraction greatly improves plasma antioxidant capacity and expression of antioxidant genes in hypercholesterolemic rats. AB - The antioxidant activities of the thymoquinone-rich fraction (TQRF) extracted from Nigella sativa and its bioactive compound, thymoquinone (TQ), in rats with induced hypercholesterolemia were investigated. Rats were fed a semipurified diet supplemented with 1% (w/w) cholesterol and were treated with TQRF and TQ at dosages ranging from 0.5 to 1.5 g/kg and 20 to 100 mg/kg body wt, respectively, for 8 weeks. The hydroxyl radical (OH(.))-scavenging activity of plasma samples collected from experimental rats was measured by electron spin resonance. The GenomeLab Genetic Analysis System was used to study the molecular mechanism that mediates the antioxidative properties of TQRF and TQ. Plasma total cholesterol and low-density-lipoprotein cholesterol levels were significantly decreased in the TQRF- and TQ-treated rats compared to untreated rats. Feeding rats a 1% cholesterol diet for 8 weeks resulted in a significant decrease in plasma antioxidant capacity, as measured by the capacity to scavenge hydroxyl radicals. However, rats treated with TQRF and TQ at various doses showed significant inhibitory activity toward the formation of OH(.) compared to untreated rats. Upon examination of liver RNA expression levels, treatment with TQRF and TQ caused the up-regulation of the superoxide dismutase 1 (SOD1), catalase, and glutathione peroxidase 2 (GPX) genes compared to untreated rats (P<0.05). In support of this, liver antioxidant enzyme levels, including SOD1 and GPX, were also apparently increased in the TQRF- and TQ-treated rats compared to untreated rats (P<0.05). In conclusion, TQRF and TQ effectively improved the plasma and liver antioxidant capacity and enhanced the expression of liver antioxidant genes of hypercholesterolemic rats. PMID- 20005292 TI - Activation of cardiac hypertrophic signaling pathways in a transgenic mouse with the human PRKAG2 Thr400Asn mutation. AB - Human mutations in PRKAG2, the gene encoding the gamma2 subunit of AMP activated protein kinase (AMPK), cause a glycogen storage cardiomyopathy. In a transgenic mouse with cardiac specific expression of the Thr400Asn mutation in PRKAG2 (TG(T400N)), we previously reported initial cardiac hypertrophy (ages 2-8 weeks) followed by dilation and failure (ages 12-20 weeks). We sought to elucidate the molecular mechanisms of cardiac hypertrophy. TG(T400N) mice showed significantly increased cardiac mass/body mass ratios up to approximately 3-fold beginning at age 2 weeks. Cardiac expression of ANP and BNP were approximately 2- and approximately 5-fold higher, respectively, in TG(T400N) relative to wildtype (WT) mice at age 2 weeks. NF-kappaB activity and nuclear translocation of the p50 subunit were increased approximately 2- to 3-fold in TG(T400N) hearts relative to WT during the hypertrophic phase. Phosphorylated Akt and p70S6K were elevated approximately 2-fold as early as age 2 weeks. To ascertain whether these changes in TG(T400N) mice were a consequence of increased AMPK activity, we crossbred TG(T400N) with TG(alpha2DN) mice, which express a dominant negative, kinase dead mutant of the AMPK alpha2 catalytic subunit and have low myocardial AMPK activity. Genetic reversal of AMPK overactivity led to a reduction in hypertrophy, nuclear translocation of NF-kappaB, phosphorylated Akt, and p70S6K. We conclude that inappropriate activation of AMPK secondary to the T400N PRKAG2 mutation is associated with the early activation of NF-kappaB and Akt signaling pathway, which mediates cardiac hypertrophy. PMID- 20005293 TI - Caprine articular, meniscus and intervertebral disc cartilage: an integral analysis of collagen network and chondrocytes. AB - Cartilage is a tissue with only limited reparative capacities. A small part of its volume is composed of cells, the remaining part being the hydrated extracellular matrix (ECM) with collagens and proteoglycans as its main constituents. The functioning of cartilage depends heavily on its ECM. Although it is known that the various (fibro)cartilaginous tissues (articular cartilage, annulus fibrosus, nucleus pulposus, and meniscus) differ from one each other with respect to their molecular make-up, remarkable little quantitative information is available with respect to its biochemical constituents, such as collagen content, or the various posttranslational modifications of collagen. Furthermore, we have noticed that tissue-engineering strategies to replace cartilaginous tissues pay in general little attention to the biochemical differences of the tissues or the phenotypical differences of the (fibro)chondrocytes under consideration. The goal of this paper is therefore to provide quantitative biochemical data from these tissues as a reference for further studies. We have chosen the goat as the source of these tissues, as this animal is widely accepted as an animal model in orthopaedic studies, e.g. in the field of cartilage degeneration and tissue engineering. Furthermore, we provide data on mRNA levels (from genes encoding proteins/enzymes involved in the synthesis and degradation of the ECM) from (fibro)chondrocytes that are freshly isolated from these tissues and from the same (fibro)chondrocytes that are cultured for 18 days in alginate beads. Expression levels of genes involved in the cross-linking of collagen were different between cells isolated from various cartilaginous tissues. This opens the possibility to include more markers than the commonly used chondrogenic markers type II collagen and aggrecan for cartilage tissue-engineering applications. PMID- 20005294 TI - Molecular characterization and bioactivity of a CXCL13 chemokine in large yellow croaker Pseudosciaena crocea. AB - A CXCL13-like chemokine cDNA was isolated from large yellow croaker (Pseudosciaena crocea) by expressed sequence tag (EST) analysis (LycCXCL13). The full-length cDNA of LycCXCL13 is 796 nucleotides (nt) encoding a protein of 97 amino acids (aa), with a putative molecular weight of 10.7 kDa. The deduced LycCXCL13 contains a 24-aa signal peptide and a 73-aa mature polypeptide, which possesses the typical arrangement of four cysteines as found in other known CXC chemokines (C(25), C(27), C(52) and C(68)). It shares 35, 36 and 39% aa sequence identities to green puffer CXCL13-like, Atlantic salmon CXCL13 and Japanese flounder CXCL13 chemokines, and 24-29% identities to CXCL13 chemokines in mammals, respectively. Phylogenetic analysis showed that LycCXCL13 is more closely related to the CXCL13 subgroup than to any other CXC chemokine subgroups. LycCXCL13 gene was constitutively expressed in all tissues examined, except for intestine. Upon induction with poly(I:C) or inactivated trivalent bacterial vaccine, LycCXCL13 gene expression was significantly up-regulated in spleen, head kidney, heart and gills at 24 h post-injection. Real-time PCR results showed that LycCXCL13 gene expression reached peak level in spleen and head kidney at 12 h after induction by poly(I:C), while its expression increased to the highest level in head kidney at 24 h or in spleen at 48 h by bacterial vaccine. Recombinant LycCXCL13 protein produced in E. coli BL21 exhibited obvious chemotaxis to the peripheral blood leucocytes (PBLs) from large yellow croaker. These results suggest that LycCXCL13 may be involved in inflammatory responses as well as homeostatic processes in large yellow croaker. PMID- 20005295 TI - The effect of G72 genotype on neural correlates of memory encoding and retrieval. AB - Polymorphisms in the G72 (also named d-amino acid oxidase activator, DAOA) gene increase the vulnerability for schizophrenia and affective psychosis. Three recent genetic neuroimaging studies showed that variation in G72 influences the brain activity in the medial temporal lobe (MTL), supporting the hypothesis that G72 might play a modulatory role on brain activity in MTL structures. In the present study we therefore investigated the effect of G72 on the neural correlates of long-term memory encoding and retrieval in a large sample of healthy subjects (n=83) using functional magnetic resonance imaging. A face encoding and a face retrieval memory task were chosen because on the one hand they specifically activate MTL structures and on the other hand they tap into memory processes that are compromised in patients with schizophrenia and affective disorder. Despite a strong a-priori hypothesis of genotype group activation differences in the MTL along with a large sample size we did neither find an effect of G72 genotype status on brain activity in the MTL nor in any other brain regions. The present data therefore do not support the view of a general modulatory role of G72 on MTL brain activity, at least not in the domain of long-term memory encoding and retrieval. Our results highlight the importance of replication studies in genetic neuroimaging. PMID- 20005296 TI - Haplotypes of catechol-O-methyltransferase modulate intelligence-related brain white matter integrity. AB - Twin studies have indicated a common genetic origin for intelligence and for variations in brain morphology. Our previous diffusion tensor imaging studies found an association between intelligence and white matter integrity of specific brain regions or tracts. However, specific genetic determinants of the white matter integrity of these brain regions and tracts are still unclear. In this study, we assess whether and how catechol-O-methyltransferase (COMT) gene polymorphisms affect brain white matter integrity. We genotyped twelve single nucleotide polymorphisms (SNPs) within the COMT gene and performed haplotype analyses on data from 79 healthy subjects. Our subjects had the same three major COMT haplotypes (termed the HPS, APS and LPS haplotypes) as previous studies have reported as regulating significantly different levels of enzymatic activity and dopamine. We used the mean fractional anisotropy (FA) values from four regions and five tracts of interest to assess the effect of COMT polymorphisms, including the well-studied val158met SNP and the three main haplotypes that we had identified, on intelligence-related white matter integrity. We identified an association between the mean FA values of two regions in the bilateral prefrontal lobes and the COMT haplotypes, rather than between them and val158met. The haplotype-FA value associations modulated nonlinearly and fit an inverted U model. Our findings suggest that COMT haplotypes can nonlinearly modulate the intelligence-related white matter integrity of the prefrontal lobes by more significantly influencing prefrontal dopamine variations than does val158met. PMID- 20005298 TI - Hierarchical rule switching in prefrontal cortex. AB - Most real-world decision-making problems involve consideration of numerous possible actions, and it is often impossible to evaluate all of them before settling on preferred strategy. In such situations, humans might explore actions more efficiently by searching only the most likely subspace of the whole action space. To study how the brain solves such action selection problems, we designed a Multi Feature Sorting Task in which the task rules defining an optimal action have a hierarchical structure and studied concurrent brain activity using it. The task consisted of two kinds of rule switches: a higher-order switch to search for a rule across different subspaces and a lower-order switch to change a rule within the same subspace. The results revealed that the left dorsolateral prefrontal cortex (DLPFC) was more active in the higher-order switching, and the right fronto-polar cortex (FPC) was significantly activated with the lower-order switching. We discuss a possible functional model in the prefrontal cortex where the left DLPFC encodes the hierarchical organization of behaviours and the right FPC maintains and updates multiple behavioural. This interpretation is highly consistent with the previous findings and current theories of hierarchical organization in the prefrontal functional network. PMID- 20005297 TI - Unsupervised statistical learning underpins computational, behavioural, and neural manifestations of musical expectation. AB - The ability to anticipate forthcoming events has clear evolutionary advantages, and predictive successes or failures often entail significant psychological and physiological consequences. In music perception, the confirmation and violation of expectations are critical to the communication of emotion and aesthetic effects of a composition. Neuroscientific research on musical expectations has focused on harmony. Although harmony is important in Western tonal styles, other musical traditions, emphasizing pitch and melody, have been rather neglected. In this study, we investigated melodic pitch expectations elicited by ecologically valid musical stimuli by drawing together computational, behavioural, and electrophysiological evidence. Unlike rule-based models, our computational model acquires knowledge through unsupervised statistical learning of sequential structure in music and uses this knowledge to estimate the conditional probability (and information content) of musical notes. Unlike previous behavioural paradigms that interrupt a stimulus, we devised a new paradigm for studying auditory expectation without compromising ecological validity. A strong negative correlation was found between the probability of notes predicted by our model and the subjectively perceived degree of expectedness. Our electrophysiological results showed that low-probability notes, as compared to high-probability notes, elicited a larger (i) negative ERP component at a late time period (400-450 ms), (ii) beta band (14-30 Hz) oscillation over the parietal lobe, and (iii) long-range phase synchronization between multiple brain regions. Altogether, the study demonstrated that statistical learning produces information theoretic descriptions of musical notes that are proportional to their perceived expectedness and are associated with characteristic patterns of neural activity. PMID- 20005299 TI - The evolutionary diversification of the Centropagidae (Crustacea, Calanoida): A history of habitat shifts. AB - The copepod family Centropagidae is widely distributed and occurs in marine, estuarine, freshwater, and inland saline settings. Molecular phylogenies based upon the 16S and 28S genes demonstrate a complex biogeographic history, involving at least five independent invasions of continental waters from the sea. The first colonization was ancient, likely into part of Gondwanaland, and resulted in an inland radiation in southern genera via both vicariance and subsequent habitat shifting among different types of continental waters. Species occupying saline lakes are nested within freshwater clades, indicating invasion of these habitats via fresh waters rather than directly from the ocean or from epicontinental seas. In contrast with the great southern clade, all of the remaining continental invasions are northern, species poor, and quite recent, perhaps even Pleistocene. Long-lived evolutionary euryhalinity, a high propensity for inland invasion, continental vicariance, and in situ radiation within single continents have all played major roles in the diversification of the centropagids. PMID- 20005300 TI - On the dynamics of nitrite, nitrate and other biomarkers of nitric oxide production in inflammatory bowel disease. AB - Nitrite and nitrate are frequently used surrogate markers of nitric oxide (NO) production. Using rat models of acute and chronic DSS-induced colitis we examined the applicability of these and other NO-related metabolites, in tissues and blood, for the characterization of inflammatory bowel disease. Global NO dynamics were assessed by simultaneous quantification of nitrite, nitrate, nitroso and nitrosyl species over time in multiple compartments. NO metabolite levels were compared to a composite disease activity index (DAI) and contrasted with measurements of platelet aggregability, ascorbate redox status and the effects of 5-aminosalicylic acid (5-ASA). Nitroso products in the colon and in other organs responded in a manner consistent with the DAI. In contrast, nitrite and nitrate, in both intra- and extravascular compartments, exhibited variations that were not always in step with the DAI. Extravascular nitrite, in particular, demonstrated significant temporal instabilities, ranging from systemic drops to marked increases. The latter was particularly evident after cessation of the inflammatory stimulus and accompanied by profound ascorbate oxidation. Treatment with 5-ASA effectively reversed these fluctuations and the associated oxidative and nitrosative stress. Platelet activation was enhanced in both the acute and chronic model. Our results offer a first glimpse into the systemic nature of DSS induced inflammation and reveal a greater complexity of NO metabolism than previously envisioned, with a clear dissociation of nitrite from other markers of NO production. The remarkable effectiveness of 5-ASA to abrogate the observed pattern of nitrite instability suggests a hitherto unrecognized role of this molecule in either development or resolution of inflammation. Its possible link to tissue oxygen consumption and the hypoxia that tends to accompany the inflammatory process warrants further investigation. PMID- 20005301 TI - LPS mediated injury to oligodendrocytes is mediated by the activation of nNOS: relevance to human demyelinating disease. AB - Loss of oligodendrocytes and the destruction of myelin form the core features of inflammatory demyelinating disease. Although many of the inflammatory and cellular mediators of tissue injury are known, recent studies have suggested an important role for nitric oxide NO and other reactive nitrogen species in oligodendrocyte injury. The human transformed oligodendrocyte cell line, MO3.13 cells, express Toll like receptor genes (TLR) genes and are activated by lipopolysaccharide (LPS). We determined the activation and consequences of neuronal nitric oxide synthase (nNOS) following stimulation with LPS in the MO3.13 cell line. Our studies show that MO3.13 cells induce nNOS following stimulation with LPS. Most importantly, these studies show a susceptibility of MO3.13 cells to NO mediated cell death by the activation of nNOS but not of inducible NOS (iNOS). MO3.13 cells show increased susceptibility to peroxynitrite mediated cellular injury to mitochondrial proteins and decreased cell survival in the presence of LPS. Our studies suggest that the presence and activation of nNOS in oligodendrocytes can directly mediate oligodendrocyte (OC) injury and reduce cell viability. PMID- 20005302 TI - Evidence for an intact polysaccharide capsule in Bordetella pertussis. AB - Polysaccharide capsules contribute to the pathogenesis of many bacteria species by providing resistance against various defense mechanisms. The production of a capsule in Bordetella pertussis, the etiologic agent of whooping cough, has remained controversial; earlier studies reported this pathogen as a capsulated microorganism whereas the recent B. pertussis genome analysis revealed the presence of a truncated capsule locus. In this work, using transmission electron microscopy and immunostaining approaches, we provide a formal evidence for the presence of an intact microcapsule produced at the surface of both laboratory strain and clinical isolates of B. pertussis. In agreement with previous studies, we found that the capsule is optimally produced in avirulent phase. Unexpectedly, the presence of the capsule was also detected at the surface of virulent B. pertussis bacteria. Consistently, a substantial transcriptional activity of the capsule operon was detected in virulent phase, suggesting that the capsular polysaccharide may play a role during pertussis pathogenesis. In vitro assays indicated that the presence of the capsule does not affect B. pertussis adherence to mammalian cells and does not further protect the bacterium from phagocytosis, complement-mediated killing or antimicrobial peptide attack. PMID- 20005303 TI - Immunomodulatory activity of the lignan 7-hydroxymatairesinol potassium acetate (HMR/lignan) extracted from the heartwood of Norway spruce (Picea abies). AB - The pharmacological profile of the lignan 7-hydroxymatairesinol (HMR/lignan, HMR) includes chemopreventive effects, antioxidant properties, and mild proestrogenic activity. The present study was devised to investigate the effects of HMR on THP 1 cells, an established model of human monocytes, and on human polymorphonuclear leukocytes (PMNs). In THP-1 cells, HMR concentration-dependently reduced LPS stimulated tumor necrosis factor (TNF)-alpha secretion in the supernatant. HMR at low, sub-muM concentrations also reduced TNF-alpha mRNA, which was however enhanced by supra-muM concentrations of HMR. In human PMNs, HMR concentration dependently reduced ROS production induced by either N-formyl-Met-Leu-Phe, phorbol myristate acetate or angiotensin II, as well as interleukin-8 production induced by either N-formyl-Met-Leu-Phe or angiotensin II. Results indicate that HMR is an effective inhibitor of both monocytic THP-1 cells and of human PMNs and warrant further studies to assess their relevance for the prevention and treatment of several conditions characterized by chronic systemic inflammation. PMID- 20005304 TI - Microinjection of serum-starved mitochondria derived from somatic cells affects parthenogenetic development of bovine and murine oocytes. AB - Microinjection of isolated mitochondria into oocytes is an effective method to introduce exogenous mitochondrial DNA. In nuclear transfer procedures in which donor cell mitochondria are transferred with nuclei into recipient oocytes; development and survival rates of reconstructed embryos may be also directly influenced by mitochondrial viability. Mitochondrial viability is dramatically affected by cell culture conditions, such as serum starvation prior to nuclear transfer. This study was conducted to examine the influence of exogenous mitochondria using bovine and mouse parthenogenetic models. Mitochondria were isolated from primary cells at confluency and after serum starvation. The bovine oocytes injected with serum-starved mitochondria showed lower rates of morula and blastocyst formation when compared to uninjected controls (P<0.05). However, the developmental rates between non-starved mitochondria injection and controls were not different (P>0.05). The murine oocytes injected with serum-starved mitochondria showed lower rates of development when compared with non-starved mitochondria and controls (P<0.01). In contrast to mitochondria transfer, ooplasm transfer did not affect murine or bovine parthenogenetic development (P>0.05). The overall results showed that injection of serum-starved mitochondria influenced parthenogenetic development of both bovine and murine oocytes. Our results illustrate that the somatic mitochondria introduction accompanying nuclei has the capacity to affect reconstructed embryo development; particularly when using serum-starved cells as donor cells. PMID- 20005305 TI - Structure-guided expansion of kinase fragment libraries driven by support vector machine models. AB - This work outlines a new de novo design process for the creation of novel kinase inhibitor libraries. It relies on a profiling paradigm that generates a substantial amount of kinase inhibitor data from which highly predictive QSAR models can be constructed. In addition, a broad diversity of X-ray structure information is needed for binding mode prediction. This is important for scaffold and substituent site selection. Borrowing from FBDD, the process involves fragmentation of known actives, proposition of binding mode hypotheses for the fragments, and model-driven recombination using a pharmacophore derived from known kinase inhibitor structures. The support vector machine method, using Merck atom pair derived fingerprint descriptors, was used to build models from activity from 6 kinase assays. These models were qualified prospectively by selecting and testing compounds from the internal compound collection. Overall hit and enrichment rates of 82% and 2.5%, respectively, qualified the models for use in library design. Using the process, 7 novel libraries were designed, synthesized and tested against these same 6 kinases. The results showed excellent results, yielding a 92% hit rate for the 179 compounds that made up the 7 libraries. The results of one library designed to include known literature compounds, as well as an analysis of overall substituent frequency, are discussed. PMID- 20005306 TI - Dissecting the role of mTOR: lessons from mTOR inhibitors. AB - Recent years have observed significant advances in our understanding of how the serine/threonine kinase target of rapamycin (TOR) controls key cellular processes such as cell survival, growth and proliferation. Consistent with its role in cell proliferation, the mTOR pathway is frequently hyperactivated in a number of human malignancies and is thus considered to be an attractive target for anti-cancer therapy. Rapamycin and its analogs (rapalogs) function as allosteric inhibitors of mTORC1 and are currently used in the treatment of advanced renal cell carcinoma. Rapamycin and its derivatives bind to the small immunophilin FKBP12 to inhibit mTORC1 signalling through a poorly understood mechanism. Rapamycin/FKBP12 efficiently inhibit some, but not all, functions of mTOR and hence much interest has been placed in the development of drugs that target the kinase activity of mTOR directly. Several novel active-site inhibitors of mTOR, which inhibit both mTORC1 and mTORC2, were developed in the last year. In this manuscript, we provide a brief outline of our current understanding of the mTOR signalling pathway and review the molecular underpinnings of the action of rapamycin and novel active-site mTOR inhibitors as well as potential advantages and caveats associated with the use of these drugs in the treatment of cancer. PMID- 20005307 TI - Cross-linking with bifunctional reagents and its application to the study of the molecular symmetry and the arrangement of subunits in hexameric protein oligomers. AB - Cross-linking with a bifunctional reagent and subsequent SDS gel electrophoresis is a simple but effective method to study the symmetry and arrangement of subunits in oligomeric proteins. In this study, theoretical expressions for the description of cross-linking patterns were derived for protein homohexamers through extension of the method used for tetramers by Hajdu et al. (1976). The derived equations were used for the analysis of cross-linking by glutardialdehyde of four protein hexamers: beef liver glutamate dehydrogenase (GDH), jack bean urease, hemocyanin from the spiny lobster Panulirus pencillatus (PpHc), Escherichia coli glutamate decarboxylase (GDC) and for analysis of published data on the cross-linking of hexameric E. coli rho by dimethyl suberimidate. Best fit models showed that the subunits in the first four proteins are arranged according to D(3) symmetry in two layers, each subunit able to cross-link to three neighboring subunits for GDH and urease, or to four for PpHc and GDC. The findings indicate a dimer-of-trimers eclipsed arrangement of subunits for GDH and urease and a trimer-of-dimers staggered one for PpHc and GDC. In rho, the subunits are arranged according to D(3) symmetry in a trimer-of-dimers ring. The conclusions from cross-linking of GDH and GDC, PpHc and rho are consistent with results from X-ray crystal structure, those for urease with findings from electron microscopy. PMID- 20005309 TI - Studies on bound water restrained by poly(2-methacryloyloxyethyl phosphorylcholine): Comparison with polysaccharide-water systems. AB - The structural change of water restrained by poly(2-methacryloyloxyethyl phosphorylcholine) (PMPC) was investigated by differential scanning calorimetry (DSC), since the biocompatibility of PMPC and related biopolymers is affected by the structure of water on the polymer surface. The phase transition behaviour of PMPC-water systems with a water content (W(c)=mass of water/mass of dry sample, gg(-1)) in the range 0-2.0 was measured in the temperature range -150 to 50 degrees C. Glass transition, cold crystallization and melting were observed. Cold crystallization, which has been suggested as an index of biocompatibility, was detected for PMPC with a W(c) in the range 0.5-0.9. The amounts of two types of bound water, non-freezing water and freezing bound water, were calculated from the melting enthalpy. The amount of non-freezing water of PMPC was approximately 0.48. It was found that the phase transition behaviour and amount of bound water of PMPC were quite similar to those of water-soluble polysaccharide electrolytes. The results indicate that the bound water, not the free water, is restrained by PMPC. PMID- 20005310 TI - Nanotextured titanium surfaces for enhancing skin growth on transcutaneous osseointegrated devices. AB - A major problem with transcutaneous osseointegrated implants is infection, mainly due to improper closure of the implant-skin interface. Therefore, the design of transcutaneous osseointegrated devices that better promote skin growth around these exit sites needs to be examined and, if successful, would clearly limit infection. Due to the success already demonstrated for orthopedic implants, developing surfaces with biologically inspired nanometer features is a design criterion that needs to be investigated for transcutaneous devices. This study therefore examined the influence of nanotextured titanium (Ti) created through electron beam evaporation and anodization on keratinocyte (skin-forming cell) function. Electron beam evaporation created Ti surfaces with nanometer features while anodization created Ti surfaces with nanotubes. Conventional Ti surfaces were largely micron rough, with few nanometer surface features. Results revealed increased keratinocyte adhesion in addition to increased keratinocyte spreading and differences in keratinocyte filopodia extension on the nanotextured Ti surfaces prepared by either electron beam evaporation or anodization compared to their conventional, unmodified counterparts after 4h. Results further revealed increased keratinocyte proliferation and cell spreading over 3 and 5days only on the nanorough Ti surfaces prepared by electron beam evaporation compared to both the anodized nanotubular and unmodified Ti surfaces. Therefore, the results from this in vitro study provided the first evidence that nano-modification techniques should be further researched as a means to possibly improve skin growth, thereby improving transcutaneous osseointegrated orthopedic implant longevity. PMID- 20005308 TI - Nutrient-dependent regulation of PGC-1alpha's acetylation state and metabolic function through the enzymatic activities of Sirt1/GCN5. AB - Mammals possess an intricate regulatory system for controlling flux through fuel utilization pathways in response to the dietary availability of particular macronutrients. Under fasting conditions, for instance, mammals initiate a whole body metabolic response that limits glucose utilization and favors fatty acid oxidation. Understanding the underlying mechanisms by which this process occurs will facilitate the development of new treatments for metabolic disorders such as type II diabetes and obesity. One of the recently identified components of the signal transduction pathway involved in metabolic reprogramming is PGC-1alpha. This transcriptional coactivator is able to coordinate the expression of a wide array of genes involved in glucose and fatty acid metabolism. The nutrient mediated control of PGC-1alpha activity is tightly correlated with its acetylation state. In this review, we evaluate how the nutrient regulation of PGC 1alpha activity squares with the regulation of its acetylation state by the deacetylase Sirt1 and the acetyltransferase GCN5. We also propose an outline of additional experimental directives that will help to shed additional light on this very powerful transcriptional coactivator. PMID- 20005311 TI - Day case surgery training for surgical trainees: a disappearing act? AB - INTRODUCTION: Over the past decade there has been considerable change to surgical training such as modernising medical careers which have raised concerns over exposure to operative experience. With the National Health Service (NHS) plan aiming for the majority of elective surgical cases to be performed as day cases we sought to assess the level of exposure modern day surgical trainees obtain in day case surgery. METHODS: An anonymous electronic questionnaire survey was completed by 100 surgical trainees in surgical training across the United Kingdom (UK) from a variety of sub-specialities. 16 questions pertinent to day case surgery exposure were answered. RESULTS: The majority of the trainees who completed the survey felt day case surgery is a vital part of their training as a surgeon. Only less than one-third of all the trainees had formal timetabled day case surgery lists. Of the 31 trainees who had scheduled day lists only 58% (n = 18) were consistently able to attend. The most common reasons for being unable to attend were rota issues and lack of encouragement from seniors. 90 trainees (90%) were not satisfied with their overall Day Case Surgery training. CONCLUSIONS: The survey reveals that the modern surgical trainee is gaining a low and inconsistent level of exposure to day case surgery despite being aware of the importance of this modality of training. An urgent review is required to ensure trainees become actively involved in day case surgery and are not missing on this vital training opportunity. PMID- 20005312 TI - Three trocar laparoscopic Roux-en-y gastric bypass: a novel technique en route to the single-incision laparoscopic approach. AB - BACKGROUND: Laparoscopic Roux-en-Y gastric bypass is the gold standard bariatric procedure. Typically, the procedure necessitates five to seven small skin incisions for trocar placement. The senior author (AA Saber) has developed a three-trocar approach for laparoscopic Roux-en-Y gastric bypass. METHODS: Sixteen patients underwent triple-incision laparoscopic Roux-en-Y gastric bypass between May 2009 and August 2009. The same surgeon performed all surgical interventions. The umbilicus was the main point of entry for all patients and the same operative technique and perioperative protocol were used in all patients. RESULTS: A total of sixteen triple-incision laparoscopic Roux-en-Y gastric bypasses were performed. The procedures were successfully performed in all patients. Mean operating time was 145.4 min. None of the patients required conversion to an open procedure. There were no mortalities or post-operative technical complications noted during the immediate post-operative period. CONCLUSION: Three trocar laparoscopic Roux-en-Y gastric bypass is safe, technically feasible and reproducible. This technique may be considered a "precursor" to single-incision laparoscopic Roux-en-Y gastric bypass. PMID- 20005313 TI - A lower costly laparoscopic appendectomy: our experience of more than 2000 cases. AB - Many authors emphasize the advantages of laparoscopic techniques in their experience, including the excellent cosmetic results, reduced postoperative pain, rapid functional recovery, lower incidence of adhesions, wound infections and believe that laparoscopic appendectomy is a reliable operation for treating acute appendicitis. Laparoscopic appendectomy has become a standard procedure in the treatment of acute appendicitis in our department since it was introduced in 2003. There are 414 patients underwent laparoscopic appendectomy and 37 patients underwent open appendectomy in our department in 2008. More than 2000 laparoscopic appendectomies were performed in our department during 2003-2009. However, open appendectomy is still the standard procedure in many Taiwan and worldwide hospitals. One of the reasons is that the operating costs incurred for laparoscopic appendectomy are higher than that of open appendectomy. We describe, herein, an access technique utilizing existing instrumentation that is reliable and easy. PMID- 20005314 TI - Simple technique for single incision transumbilical laparoscopic appendectomy. AB - INTRODUCTION: Acute appendicitis is one of the most commonly encountered surgical problems in everyday practice. With the recent increase in popularity of single incision laparoscopic surgery (SILS), several techniques for SILS appendectomy have already been described. We herein describe our own simplified technique for single incision transumbilical laparoscopic appendectomy. MATERIALS AND METHODS: From December 2008 to August 2009, a total of 26 patients consented for single incision laparoscopic appendectomy for acute appendicitis. Preoperative, intraoperative and postoperative data were collected. RESULTS: Our technique was successful in 19 out of 26 (73.1%) patients, while seven patients required the placement of additional trocars. None of the patients needed conversion to an open approach. Mean OR time was 45.9 min. The mean length of stay was 1.1 day. Only one patient had developed postoperative umbilical wound infection. CONCLUSION: Our technique for single incision laparoscopic transumbilical appendectomy is safe feasible, and reproducible. Prospective randomized studies comparing the single incision laparoscopic approach with its conventional multiport counterpart are necessary to confirm the conclusions of our early experience. PMID- 20005316 TI - Increased cartilage type II collagen degradation in patients with osteogenesis imperfecta used as a human model of bone type I collagen alterations. AB - OBJECTIVE: We investigated whether cartilage degradation is altered in adult patients with mild osteogenesis imperfecta (OI) used as a human model of bone type I collagen-related osteoarthritis (OA). PATIENTS AND METHODS: Sixty-four adult patients with OI (39% women, mean age+/-SD: 37+/-12 years) and 64 healthy age-matched controls (54% women, 39+/-7 years) were included. We also compared data in 87 patients with knee OA (73% women, 63+/-8 years, mean disease duration: 6 years) and 291 age-matched controls (80% women, 62+/-10 years). Urinary C terminal cross-linked telopeptide of type II collagen (CTX-II), a marker of cartilage degradation, urinary helical peptide of type I collagen (Helix-I), a marker of bone resorption, and the urinary ratio between non isomerised/isomerised (alpha/beta CTX-I) type I collagen C-telopeptide, a marker of type I collagen maturation, were measured. RESULTS: Patients with OI had CTX II levels similar to those of subjects with knee OA (p=0.89; mean+/-SEM; 460+/-57 ng/mmol Cr for OI group and 547+/-32 ng/mmol Cr for OA group) and significantly higher than both young (144+/-7.8 ng/mmol Cr, p<0.0001) and old controls (247+/-7 ng/mmol Cr, p<0.0001). In patients with OI, increased Helix-I (p<0.0001) and alpha/beta CTX-I (p=0.0067) were independently associated with increased CTX-II and together explained 26% of its variance (p< 0.0001). In patients with knee OA, increased levels of alpha/beta CTX-I ratio were also associated with higher CTX II levels. CONCLUSION: Adult patients with OI or knee OA are characterized by increased cartilage type II collagen degradation, which is associated with increased type I collagen degradation for OI and lower type I collagen maturation for both OI and OA. These data suggest that both quantitative and qualitative alterations of bone type I collagen metabolism are involved in increased cartilage degradation in patients with OI or knee OA. PMID- 20005315 TI - Quality of diet and potential renal acid load as risk factors for reduced bone density in elderly women. AB - BACKGROUND: Bone mineral density (BMD) may be influenced by the general dietary pattern and the potential renal acid load (PRAL). METHODS: We compared the dietary intake (estimated using the European Prospective Investigation into Cancer and nutrition questionnaire) of 497 community-living women (60 years of age and older) grouped according to tertiles of baseline total, trabecular and cortical BMD estimated using tibial peripheral quantitative computed tomography (pQCT), and of BMD variation over 6 years. RESULTS: None of the nutrients taken into account nor PRAL was associated with total BMD, with the exception that the intake of polyunsaturated fatty acids (PUFA) was slightly higher among women with the highest total BMD. Similar results were found for trabecular BMD. Cortical BMD was associated with serum 25-OH vitamin D (38.8, 43.2, and 49.5 nmol/L in the first, second, and third tertiles, respectively; P=0.042). In the longitudinal analysis, a lower BMI was associated with greater loss of total BMD, while lower serum 25-OH vitamin D at baseline was associated with smaller loss of cortical BMD. CONCLUSIONS: We found no relationship between dietary acid load and BMD. We also confirmed the role of well-recognized risk factor for osteoporosis. PMID- 20005317 TI - Knowledge and acceptability of the HPV vaccine by school children and their parents in Birmingham. AB - Previous research indicates that knowledge about the HPV vaccine amongst the general public is inadequate [1-3]. However, since the introduction of the NHS vaccination programme for girls aged 12-13 from autumn 2008 awareness may have increased. We aimed to assess their knowledge about the vaccine, their likelihood of vaccine uptake and whether these were affected by demographic variables such as gender and ethnicity. Questionnaires which were designed to test levels of knowledge about the vaccination programme were distributed to a sample of 12-13 year-old boys and girls and their parents from three schools in Birmingham. In total, 568 questionnaires were distributed, with 434 responses (76.4%). The results suggest that knowledge is still poor, particularly in subgroups such as males, those of non-white ethnicity and children. Furthermore, we found many misconceptions exist amongst the participants, which could have significant implications for information provision and the targeting of future education programmes. PMID- 20005318 TI - Amino acid residues 196-225 of LcrV represent a plague protective epitope. AB - LcrV, a protein that resides at the tip of the type III secretion needles of Yersinia pestis, is the single most important plague protective antigen. Earlier work reported monoclonal antibody MAb 7.3, which binds a conformational epitope of LcrV and protects experimental animals against lethal plague challenge. By screening monoclonal antibodies directed against LcrV for their ability to protect immunized mice against bubonic plague challenge, we examined here the possibility of additional protective epitopes. MAb BA5 protected animals against plague, neutralized the Y. pestis type III secretion pathway and promoted opsonophagocytic clearance of bacteria in blood. LcrV residues 196-225 were necessary and sufficient for MAb BA5 binding. Compared to full-length LcrV, a variant lacking its residues 196-225 retained the ability of eliciting plague protection. These results identify LcrV residues 196-225 as a linear epitope that is recognized by the murine immune system to confer plague protection. PMID- 20005319 TI - Analytical methods for the determination of arsenosugars--a review of recent trends and developments. AB - Arsenic-containing carbohydrates, generally termed arsenosugars, have been the subject of increasing analytical interest in arsenic speciation analysis. The present review gives an overview concerning achievements and trends in the field of instrumental analysis of arsenosugars. The typical experimental approaches for sample pre-treatment, extraction, separation and detection are discussed. Current possibilities and limitations of modern instrumental techniques are pointed out. PMID- 20005320 TI - A systematic approach to the accurate quantification of selenium in serum selenoalbumin by HPLC-ICP-MS. AB - In this paper, two different methods are for the first time systematically compared for the determination of selenium in human serum selenoalbumin (SeAlb). Firstly, SeAlb was enzymatically hydrolyzed and the resulting selenomethionine (SeMet) was quantified using species-specific isotope dilution (SSID) with reversed phase-HPLC (RP-HPLC) hyphenated to (collision/reaction cell) inductively coupled plasma-quadrupole mass spectrometry (CRC ICP-QMS). In order to assess the enzymatic hydrolysis yield, SeAlb was determined as an intact protein by affinity HPLC (AF-HPLC) coupled to CRC ICP-QMS. Using this approach, glutathione peroxidase (GPx) and selenoprotein P (SelP) (the two selenoproteins present in serum) were also determined within the same chromatographic run. The levels of selenium associated with SeAlb in three serum materials, namely BCR-637, Seronorm level 1 and Seronorm level 2, obtained using both methods were in a good agreement. Verification of the absence of free SeMet, which interferes with the SeAlb determination (down to the amino acid level), in such materials was addressed by analyzing the fraction of GPx, partially purified by AF-HPLC, using RP-HPLC (GPx only) and size exclusion-HPLC (SE-HPLC) coupled to CRC ICP-QMS. The latter methodology was also used for the investigation of the presence of selenium species other than the selenoproteins in the (AF-HPLC) SelP and SeAlb fractions; the same selenium peaks were detected in both control and BCR-637 serum with a difference in age of ca. 12 years. It is also for the first time that the concentrations of selenium associated with SeAlb, GPx and SelP species in such commercially available serums (only certified or having indicative levels of total selenium content) are reported. Such indicative values can be used for reference purposes in future validation of speciation methods for selenium in human serum and/or inter-laboratory comparisons. PMID- 20005321 TI - Deconvolution of liquid scintillation alpha spectra of mixtures of uranium and radium isotopes. AB - A method for the determination of uranium and radium isotopes in water samples is proposed. Liquid scintillation techniques were used for collecting alpha spectra, which were then analyzed by fitting the alpha peaks with overlapping Gaussians. The analysis can quantify the observed isotopes with accuracy depending on the activity of each isotope. In order to simulate the peaks with Gaussian normal distribution functions, the centroid of each peak as well as the full width at half maximum (FWHM) are required, as they depend on the quenching of the sample. For this purpose, samples with known activities of 226Ra and its decay products and also of the uranium isotopes 238U and 234U, at various quenching levels, were used to establish the correlation of the peaks' shift with the quench effect. In addition, the correlation of the FWHM with the centroid of a peak was determined, using the same procedure. Following the above analysis technique, an average of 97+/-2% of detection efficiency and a lower limit of detection of 8.2 mBq kg(-1) for alpha isotopes were achieved. PMID- 20005322 TI - Canonical Measure of Correlation (CMC) and Canonical Measure of Distance (CMD) between sets of data. Part 3. Variable selection in classification. AB - In multivariate regression and classification issues variable selection is an important procedure used to select an optimal subset of variables with the aim of producing more parsimonious and eventually more predictive models. Variable selection is often necessary when dealing with methodologies that produce thousands of variables, such as Quantitative Structure-Activity Relationships (QSARs) and highly dimensional analytical procedures. In this paper a novel method for variable selection for classification purposes is introduced. This method exploits the recently proposed Canonical Measure of Correlation between two sets of variables (CMC index). The CMC index is in this case calculated for two specific sets of variables, the former being comprised of the independent variables and the latter of the unfolded class matrix. The CMC values, calculated by considering one variable at a time, can be sorted and a ranking of the variables on the basis of their class discrimination capabilities results. Alternatively, CMC index can be calculated for all the possible combinations of variables and the variable subset with the maximal CMC can be selected, but this procedure is computationally more demanding and classification performance of the selected subset is not always the best one. The effectiveness of the CMC index in selecting variables with discriminative ability was compared with that of other well-known strategies for variable selection, such as the Wilks' Lambda, the VIP index based on the Partial Least Squares-Discriminant Analysis, and the selection provided by classification trees. A variable Forward Selection based on the CMC index was finally used in conjunction of Linear Discriminant Analysis. This approach was tested on several chemical data sets. Obtained results were encouraging. PMID- 20005324 TI - The application of digital image recognition to the analysis of two-dimensional fingerprints. AB - High-performance liquid chromatography (HPLC) fingerprint has been commonly used in the quality control and assessment of herbal medicines, and two-dimensional (2D) fingerprint obtained by means of HPLC-diode array detector (HPLC/DAD) can provide higher reliability. In this paper, an approach to the analysis of the 2D HPLC/DAD fingerprints, which was based on digital image recognition techniques, was developed for the first time. First, wavelet transform was employed to eliminate noise signal in the 2D fingerprint, and then the 2D fingerprint was converted into grayscale image. Second, several features of the image were calculated, and hierarchical clustering. This approach was applied to the qualitative analysis of the different samples of coptis chinensis, and the clustering result of samples was all highly consistent with the real situation. Based on the densities in grayscale image, three components in standard samples were quantitative analyzed, and the obtained correlation coefficients between concentration and grayscale density were more than 0.999. Our study indicated that the analysis of the 2D HPLC/DAD fingerprint was successful based on the idea and techniques of digital image recognition techniques, and this proposed approach provided a new pathway for the analysis of two-dimensional spectrums. PMID- 20005323 TI - Evaluation of the influence of proline, hydroxyproline or pyrrolidine in the presence of sodium nitrite on N-nitrosamine formation when heating cured meat. AB - N-nitrosamines are meant to be probable or possible carcinogenic components, possibly formed out of a reaction between nitrite and N-containing substances such as amino acids and secondary amines. Nitrite is often used for processing meat products because of its colouring and antimicrobial properties. During this experimental setup, the influence of proline, hydroxyproline or pyrrolidine on N nitrosamine formation in meat samples was evaluated. The N-nitrosamines concentrations were measured with gas chromatography-thermal energy analyzer. Only the concentrations of N-nitrosodimethylamine and N-nitrosopyrrolidine were found above the limit of detection in a number of tested experimental conditions. The concentration of these two N-nitrosamines was modelled as a function of temperature and nitrite concentration for different situations (presence or absence of added natural N-containing meat components). It could be concluded that proline and pyrrolidine promoted the formation of N-nitrosopyrrolidine. It could also be confirmed that the higher the temperature of the meat processing procedure and the higher the sodium nitrite amounts added, the higher were the yields of the respective N-nitrosamines. PMID- 20005325 TI - A multivariate multianalyte screening method for sulfonamides in milk based on front-face fluorescence spectroscopy. AB - Screening methods are used to detect the presence of a substance or class of substances at the level of interest and are specifically designed to avoid false compliant results. They should allow the running of a high number of samples per day at a low cost under routine conditions. In this work, a rapid and simple method for the screening of six sulfonamides (sulfadiazine, SD; sulfamerazine, SMR; sulfamethazine, SMT; sulfachloropyridazine, SCP; sulfathiazole, STZ and sulfamethoxazole, SMO) in milk samples is proposed and assessed according to the criteria required by the European Regulation, Decision 2002/657/EC. The method is based on modelling front-face fluorescence emission spectra by means of partial least squares class modelling (PLS-CM). The milk samples are pre-treated with a single easy step of derivatization with fluorescamine. After confirming that the method has equal analytical sensitivity for all the six sulfonamides, it is established that the multivariate analytical sensitivity at 100 microg L(-1) is 37.5 microg L(-1) when analysing a mixture of six sulfonamides added to different brands of milk and measured in different days. In addition, the method is applied to samples from 11 commercial brands of milk. For beta=0.05, threshold value established by the Decision 2002/657/EC for this method, the probability of false non-compliance, alpha, is equal to 0.17, allowing the suitable screening of these six sulfonamides. PMID- 20005326 TI - Electrochemical investigations of corticosteroid isomers--testosterone and epitestosterone and their simultaneous determination in human urine. AB - Voltammetric investigation of two corticoid isomers--testosterone and epitestosterone has been carried out at bare and single-wall carbon nanotubes (SWNT)-modified edge plane pyrolytic graphite electrode (EPPGE). Square wave voltammetry (OSWV) has been used for the simultaneous determination of isomeric steroids. The reduction of the two isomers occurred in a pH dependent, 2e, 2H+ process and well-defined voltammetric peaks were observed. Under the optimum experimental conditions, linear calibration curves are obtained within the concentration range 5-1000 nM for both the steroids with the limit of detection 2.8 x 10(-9) and 4.1 x 10(-9) M for testosterone and epitestosterone respectively. The developed protocol is successfully implemented for the analysis of both the compounds in the urine samples of normal subjects as well as in patients undergoing treatment with testosterone. The results obtained from the proposed voltammetric method were also compared with HPLC analysis and found to be similar. PMID- 20005327 TI - Evaluation of non-specific binding suppression schemes for neutravidin and alkaline phosphatase at the surface of reticulated vitreous carbon electrodes. AB - Non-specific binding (NSB) of high-molecular-weight proteins onto electrode surfaces can complicate the application of electroanalytical techniques to clinical and environmental research, particularly in biosensor applications. We present herein various strategies to modify the surface of reticulated vitreous carbon (RVC) electrodes to suppress non-specific binding of biomolecules onto its surface. Non-specific binding and specific binding (SB) of two enzyme conjugates, neutravidin-alkaline phosphatase (NA-ALP) and biotinylated alkaline phosphatase (B-ALP), and also neutravidin itself, were studied using hydroquinone diphosphate (HQDP) as an enzyme substrate for ALP inside the pores of RVC electrodes that had been subjected to various modification schemes. The extent of NSB and SB of these biomolecules inside RVC pores was assessed by measuring the initial rate of generation of an electroactive product, hydroquinone (HQ), of the enzyme catalyzed reaction, using linear scan voltammetry (LSV) for HQ detection. Electrodes functionalized with phenylacetic acid and poly(ethylene glycol) (PEG) showed low NSB and high SB (when biotin capture ligands were included in the modification scheme) in comparison with unmodified electrodes and RVC electrodes modified in other ways. A simple sandwich bioassay for neutravidin was performed on the RVC electrode with the lowest NSB. A concentration detection limit of 52+/ 2 ng mL(-1) and an absolute detection limit of 5.2+/-0.2 ng were achieved for neutravidin when this assay was performed using a 100 microL sample size. PMID- 20005328 TI - Electroanalytical study of SYBR Green I and ethidium bromide intercalation in methylated and unmethylated amplicons. AB - This work involves the electrochemical study of the interaction of SYBR Green I (SG) with native DNA using differential pulse voltammetry at a carbon paste electrode (CPE) and alternating current voltammetry at a hanging mercury drop electrode (HMDE). At the CPE the peak current intensity at 1.0 V decreased by increasing the concentration of SG. At the HMDE, a decrease in the current intensity of the DNA peak at -1.2 V was also observed by increasing the concentration of SG. These results electrochemically confirmed that SG intercalates within the DNA double helix and changes its conformation. Through the present work the differentiation of differently methylated analytes was achieved by application of alternative current and differential pulse voltammetric techniques. Amplicons (PCR products) corresponding to the GC-rich p53 exon 5 containing cytosine and its methylated analogue, synthesized by substituting 60% of cytosine by 5-methyl-cytosine, were amplified and investigated electrochemically in the presence of SG and ethidium bromide (EtBr) by differential pulse voltammetry. Considerable peak current differences were observed in the presence of SG and EtBr for unmethylated exon 5 vs. methylated. Therefore, both SG and EtBr could serve as electrochemical probes for identifying different DNA conformations. PMID- 20005329 TI - The use of stir bar sorptive extraction--a potential alternative method for the determination of furan, evaluated using two example food matrices. AB - A comparison is made between static headspace analysis and stir bar sorptive extraction (SBSE) for the quantitative determination of furan. The SBSE technique was optimised and evaluated using two example food matrices (coffee and jarred baby food). The use of the SBSE technique in most cases, gave comparable results to the static headspace method, using the method of standard additions with d4 labelled furan as an internal standard. Using the SBSE method, limits of detection down to 2 ng g(-1) were achieved, with only a 1 h extraction. The method was performed at ambient temperatures, thus eliminating the possibility of formation of furan during extraction. PMID- 20005330 TI - Detection and identification of hydrophilic selenium compounds in selenium-rich yeast by size exclusion-microbore normal-phase HPLC with the on-line ICP-MS and electrospray Q-TOF-MS detection. AB - Normal-phase HPLC and hydrophilic interaction HPLC (HILIC) were investigated for the separation of selenometabolites in a water extract of Se-rich yeast prior to their detection by ICP-MS and identification by electrospray MS/MS. The targeted fraction was a low-abundant fraction co-eluting with salt and sulfur analogues in size-exclusion chromatography which has so far been inaccessible to Se speciation studies. The optimization of the separation conditions resulted in the highest separation efficiency when HILIC was used and elution was carried out isocratically with a low concentration ammonium acetate buffer (1 mM ammonium acetate/10 mM acetic acid) in 80% acetonitrile. Out of 15 peaks observed with the Se-specific ICP-MS detection 12 was identified by electrospray Q-TOF MS/MS (2,3 dihydroxypropionyl (DHP)-Se-methylselenocysteine [M+H]+: 272, Se-methyl-gamma glutamyl-selenocysteinylglycine dioxide [M+H]+: 402, gamma-glutamyl-Se methylselenocysteine [M+H]+: 313; isomers of gamma-glutamylselenocystathionine [M+H]+: 400; Se-methyl-selenoglutathione [M+H]+: 370, isomers of N acetylselenocystathionine [M+H]+: 313, 2,3-DHP-selenohomolanthionine [M+H]+: 373, isomers of 2,3-DHP-selenocystathionine [M+H]+: 359, 2,3-DHP-selenolanthionine [M+H]+: 345 and selenohomolanthionine [M+H]+: 285). PMID- 20005331 TI - A general-purpose baseline estimation algorithm for spectroscopic data. AB - A common feature of many modern technologies used in proteomics--including nuclear magnetic resonance imaging and mass spectrometry--is the generation of large amounts of data for each subject in an experiment. Extracting the signal from the background noise, however, poses significant challenges. One important part of signal extraction is the correct identification of the baseline level of the data. In this article, we propose a new algorithm (the "BXR algorithm") for baseline estimation that can be directly applied to different types of spectroscopic data, but also can be specifically tailored to different technologies. We then show how to adapt the algorithm to a particular technology- matrix-assisted laser desorption/ionization Fourier transform ion cyclotron resonance mass spectrometry--which is rapidly gaining popularity as an analytic tool in proteomics. Finally, we compare the performance of our algorithm to that of existing algorithms for baseline estimation. The BXR algorithm is computationally efficient, robust to the type of one-sided signal that occurs in many modern applications (including NMR and mass spectrometry), and improves on existing baseline estimation algorithms. It is implemented as the function baseline in the R package FTICRMS, available either from the Comprehensive R Archive Network (http://www.r-project.org/) or from the first author. PMID- 20005332 TI - Characterisation of free and glycosidically bound odourant compounds of Aragonez clonal musts by GC-O. AB - To evaluate the potential aroma of Aragonez clonal red musts, several free and glycosidically bound odourant compounds were extracted. Then, the gas chromatography-olfactometry (GC-O) posterior intensity method was used to evaluate their odour intensity and the compounds were identified by gas chromatography-mass spectrometry (GC-MS). A group of eight sniffers evaluated free and bound fractions of Aragonez musts and perceived forty-three and twenty two odourant peaks respectively. Furaneol (burnt sugar, candy-cotton) and vanillin (vanilla, sweet) were identified in both free and bound fractions of Aragonez musts, indicating their grape-derived origin. Principal component analysis (PCA) was applied to the posterior intensity method data and a relationship between the different odourant compound variables and the free fractions was established. Two principal components (PCs) were found which together explained 100% of the total variance. A large number of potentially important but yet unknown odourants was detected by the GC-O analysis. PMID- 20005333 TI - Construction of supported lipid membrane modified piezoelectric biosensor for sensitive assay of cholera toxin based on surface-agglutination of ganglioside bearing liposomes. AB - A novel piezoelelctric biosensor has been developed for cholera toxin (CT) detection based on the analyte-mediated surface-agglutination of ganglioside (GM1)-functionalized liposomes. To achieve a CT-specific agglutination at the surface, the gold electrode is modified by a GM1-functionalized supported lipid membrane via spontaneous spread of the liposomes on a self-assembled monolayer of a long-chain alkanethiol. In the presence of CT, the GM1-incorporated liposomes in assay medium will rapidly specifically agglutinate at the electrode surface through the binding of CT to GM1 on the electrode surface and the liposome interface. This results in an enormous mass loading on the piezoelelctric crystal as well as a significant increase of density and viscosity at the interface, thereby generating a decrease in frequency of the piezoelelctric crystal. The combination of mass loading with interfacial change in the surface-agglutination reaction allows the developed piezoelelctric biosensor to show substantial signal amplification in response to the analyte CT. The detection limit can be achieved as low as 25 ng mL(-1) CT. This is the first demonstration on CT detection based on specific surface-agglutination of GM1-modified liposomes. The supported lipid layer based sensing interface can be prepared readily and renewably, making the developed technique especially useful for simple, reusable and sensitive determination of proteins. PMID- 20005335 TI - Forty years of publication of transplantation proceedings-the third decade: the expansion of the enterprise. PMID- 20005334 TI - A sensitive immunosorbent bio-barcode assay combining PCR with icELISA for detection of gonyautoxin 2/3. AB - In the current study, we developed a nanosphere bio-barcode technology to detect trace gonyautoxin 2/3 (GTX 2/3). GTX 2/3-glucose oxidase (GOX) conjugates were first prepared as the coating antigen in a periodate reaction. Subsequently, gold nanoparticles (NP) dual-labeled with anti-GTX 2/3 monoclonal antibodies (Mab) and DNA oligonucleotides were synthesized via a one-step preparation method. Combining PCR with indirect competitive ELISA (icELISA), a novel immunosorbent bio-barcode assay was established utilizing the Mab-NP-dsDNA complex to convert enzymatic signals to DNA signals. Importantly, the limit of detection of the method was lower than 0.74 microg mL(-1). Thus, the immunosorbent bio-barcode assay is a rapid and high-throughput screening tool to detect GTX 2/3 in aquatic products. PMID- 20005336 TI - Current surgical management of hilar and intrahepatic cholangiocarcinoma: the role of resection and orthotopic liver transplantation. AB - Cholangiocarcinoma (CCA) is a rare but devastating malignancy that presents late, is notoriously difficult to diagnose, and is associated with a high mortality. Surgical resection is the only chance for cure or long-term survival. The treatment of CCA has remained challenging because of the lack of effective adjuvant therapy, aggressive nature of the disease, and critical location of the tumor in close proximity to vital structures such as the hepatic artery and the portal vein. Moreover, the operative approach is dictated by the location of the tumor and the presence of underlying liver disease. During the past 4 decades, the operative management of CCA has evolved from a treatment modality that primarily aimed at palliation to curative intent with an aggressive surgical approach to R0 resection and total hepatectomy followed by orthotopic liver transplantation. PMID- 20005337 TI - Hand-assisted laparoscopic living-donor nephrectomy versus open surgery: evaluation of surgical trauma and late graft function in 82 patients. AB - OBJECTIVE: We evaluated and quantified surgical trauma and late graft function in cases of hand-assisted laparoscopic living-donor nephrectomy (HALLDN) versus open living-donor nephrectom (OLDN). METHODS: This study is a retrospective nonrandomized single-center analysis. Between 1995 and January 2008, 82 patients with end-stage renal disease received kidney transplantations from living donors. Open living-donor nephrectomy was performed in 37 donors, and 45 underwent laparoscopic hand-assisted nephrectomy. Demographic data and perioperative and postoperative data, such as markers of acute phase (C-reactive protein; serum amyloid A) and biochemical markers of glomerular filtration (serum creatinine, serum cystatin C), were compared at serial time points. RESULTS: The mean operative times for HALLDN and OLDN were 165 min and 195 min, respectively. The average warm ischemia time was 45 seconds for laparoscopy and 87 seconds for open surgery. The evaluation of acute phase markers demonstrated a minimally invasiven nature of laparoscopy, with same late graft function compared with open surgery. CONCLUSION: When the surgery was performed by experienced surgeons, hand-assisted living- donor nephrectomy showed shorter operative and warm ischemia times than open surgery, offering at least the same functional results and decreasing surgical complications compared with a completely laparoscopic technique. PMID- 20005338 TI - Polyglactin tie added to nonabsorbable polymer locking clips to control artery in laparoscopic living donor nephrectomy: better safe than sorry. AB - As Teleflex Medical contraindicates the use of Hem-O-Lok clips in laparoscopic living donor nephrectomy (LLDN), we report the feasibility of a simple surgical artifice that aims to preserve the advantages of lockable clips with increased safety while respecting the manufacturer's legal recommendations. Since January 2009, a polyglactin-0 tie was placed on the renal artery in addition to the two usual Hem-O-Lok clips in LLDN at our institution (n = 10) using a pre-tied loop suture (Endoloop ligature, Ethicon) placed on the artery stump, proximally to the aorta, after kidney removal. This artifice increased operating time of 65 seconds (range, 35-85 seconds) with no modification of warm ischemia time and led to visually decreased aortic pulsation transmitted to the clips. Without evidence of increased safety, we assume that this ruse may protect surgeons from prosecution in cases of clip displacement. It certainly decreases the risk of clip slippage and should be considered as a cheap, easy artifice to reduce the already low-risk of hemorrhage in LLDN. PMID- 20005339 TI - Renal cysts in living donor kidney transplantation: long-term follow-up in 25 patients. AB - INTRODUCTION: The acceptance of a living donor kidney bearing cysts might implicate complications after the transplantation due to the natural history of renal cysts. We have presented our experience with transplantation of living donor kidneys containing cysts but not polycystic disease. PATIENTS AND METHODS: We retrospectively reviewed donor and recipient records of all living kidney transplants performed between January 1997 and April 2008. We analyzed serum creatinine and urea levels, as well as ultrasound scans concerning cyst size and morphology at hospital discharge as well as at 12 and 24 months after transplantation. RESULTS: Among 268 living kidney transplantations, we noted 25 donors with renal cysts. In the computed tomography scan reports, 19 kidneys were described to show a single and six, multiple cysts. The size of 10 single cysts was <5 mm; the other nine were a mean of 17.33 mm. Two of the multiple cyst kidneys had lesions <5 mm; in four kidneys, the mean cyst size was 27.25 mm. The renal function of the recipients was normal or almost normal at discharge with a tendency to lower levels at 12 and 24 months after transplantation. Ultrasound revealed changes in cyst diameter among 6/23 kidneys; the mean diameter increased after 12 months, namely, 8.25 mm to 11.5 mm after 24 months. The subgroup of patients with enlarged cysts showed creatinine and urea levels slightly higher than in the entire group. No aspects of malignancy were found, according to the Bosniak and Israel classification system. One suspicious cyst was tomographically confirmed to be hemorrhagic without any need for treatment. None of the living donors had any problems related to the donor nephrectomy or a need for dialysis due to renal insufficiency in the long term. In addition, the living donors who had even beforehand cystic lesions in their contralateral nonremoved kidney at the time of transplantation did not show complications upon follow-up. CONCLUSIONS: In our study, 25 living donor kidneys carried cysts. Neither cyst related complications nor dysfunction of the transplanted organs occurred. An unroofing or excision of the cyst was generally not necessary. Regular ultrasound scans and optional computed tomography scans are recommended for follow-up. Based on this experience, we concluded that kidneys presenting cystic diseases should be considered to be suitable for transplantation without a hazard to the recipients, thus extending the pool of organs. PMID- 20005340 TI - Estimation of standard liver volume in Chinese adult living donors. AB - AIM: To determine a formula predicting the standard liver volume based on body surface area (BSA) or body weight in Chinese adults. MATERIALS AND METHODS: A total of 115 consecutive right-lobe living donors not including the middle hepatic vein underwent right hemi-hepatectomy. No organs were used from prisoners, and no subjects were prisoners. Donor anthropometric data including age, gender, body weight, and body height were recorded prospectively. The weights and volumes of the right lobe liver grafts were measured at the back table. Liver weights and volumes were calculated from the right lobe graft weight and volume obtained at the back table, divided by the proportion of the right lobe on computed tomography. By simple linear regression analysis and stepwise multiple linear regression analysis, we correlated calculated liver volume and body height, body weight, or body surface area. RESULTS: The subjects had a mean age of 35.97 +/- 9.6 years, and a female-to-male ratio of 60:55. The mean volume of the right lobe was 727.47 +/- 136.17 mL, occupying 55.59% +/- 6.70% of the whole liver by computed tomography. The volume of the right lobe was 581.73 +/- 96.137 mL, and the estimated liver volume was 1053.08 +/- 167.56 mL. Females of the same body weight showed a slightly lower liver weight. By simple linear regression analysis and stepwise multiple linear regression analysis, a formula was derived based on body weight. All formulae except the Hong Kong formula overestimated liver volume compared to this formula. CONCLUSIONS: The formula of standard liver volume, SLV (mL) = 11.508 x body weight (kg) + 334.024, may be applied to estimate liver volumes in Chinese adults. PMID- 20005341 TI - Attitudes and behaviours of students from the faculty of theology regarding organ donation: a study from Turkey. AB - The aim of this study was to investigate the knowledge and attitude of students from the Faculty of Theology of Erciyes University regarding organ donation. This study comprising all students (n = 264) showed that 51.6% of subjects to the kidney is an organ that may be donated; other organs were less known. 16.5% of the students thought that organ donation is not in accord with Islamic beliefs; 22.0% thought that it is permitted in Islam for Muslims to donate to non-Muslims, and 23.6% were willing to accept organs from non-Muslims. 23.6% of the students were willing to donate their organs, whereas 57.3% were undecided. None of the students had an organ donation card. Among students who did not consider donation or were undecided, 16.5% stated that it was "religiously inappropriate" and 13.3% stated that they did not "approve the loss of body integrity." Students declared that they had little knowledge regarding organ/tissue donation: 67.9% about the religious aspect, 78.9% about the legal aspect, and 80.5% about the scientific aspect. Only 24.6% of the group noted school education as their source of information, with 51.2% stating that they had been questioned about organ donation by society. With this study, we concluded that the student's knowledge regarding organ donation was not sufficient. PMID- 20005342 TI - Effects of delayed rapamycin treatment on renal fibrosis and inflammation in experimental ischemia reperfusion injury. AB - Ischemia reperfusion injury (IRI) has long-term sequelae on kidney allograft function. Early initiation of rapamycin can retard surgical wound healing and recovery from IRI. In contrast, rapamycin may paradoxically retard long-term fibrotic effects of kidney IRI. We, therefore, hypothesized that delayed initiation of rapamycin after kidney ischemia, started after the initial week of wound healing, would decrease the long-term inflammation and fibrosis caused by IRI. C57BL/6 male mice were subjected to either 45 or 60 minutes of unilateral kidney ischemia or a sham operation. Mice were given rapamycin (subcutaneous, 1.5 mg/kg/d) or vehicle starting at 1 week after IRI surgery for 3 weeks. Urine albumin excretion, kidney histology, and kidney cytokine proteins were examined at 4 weeks after surgery. The 3-week treatment course of rapamycin significantly reduced body weight gain in all 3 groups and reduced postischemic kidney weight in both the 45- and 60-minute ischemia groups, but unexpectedly increased urine albumin excretion in all rapamycin-treated sham or IRI mice compared with vehicle treated mice. Rapamycin treatment showed minimal effects on postischemic kidney fibrosis with variable effects on various cytokine/chemokine protein expressions, namely, decreasing interleukin (IL)-1alpha, IL-6, tumor necrosis factor (TNF) alpha, and regulated on activation normal T cell expressed and secreted (RANTES) while increasing IL-4, keratinocyte-derived chemokine (KC), macrophage inflammatory protein (MIP-1alpha), and IL-10 in the ischemic kidney. These data demonstrated that rapamycin reduced mouse body weight and ischemic kidney weight, while increasing urinary albumin excretion. Delayed initiation of rapamycin after IRI had a minimal effect on renal fibrosis and mixed effects on proinflammatory mediator production. These data do not support delayed initiation of rapamycin after IRI to attenuate IRI-induced progressive fibrosis and inflammation, and They raise further caution regarding rapamycin and albuminuria. PMID- 20005343 TI - Comparison of preservation solutions for washout of kidney grafts: an experimental study. AB - OBJECTIVE: The impact of different preservation solutions for washout of kidney grafts was evaluated regarding temperature, kidney weight, remaining red blood cells (RBCs) and histological evaluation after ex vivo washout using 500 mL cold preservation solution at 4 degrees C followed by 24 hours cold storage (CS). METHODS: Kidneys retrieved from Landrace pigs (20-30 kg) were immediately washed (warm ischemic time 0 min [WIT 0]), using 500 mL cold University of Wisconsin solution (UW), histidine-tryptophan-ketoglutarate (HTK), or Polysol (PS) followed by 24 hours, CS. Also, kidneys were retrieved after a WIT of 30 minutes followed by washout using HTK or PS. RESULTS: After washout, the weight of kidneys washed out with HTK had increased, whereas that of organs in the UW or PS group had decreased. After washout with UW, the core temperature of WIT 0 kidneys was lower than that with HTK. The time needed for washout using 500 mL solution was shorter using PS compared with HTK for both WIT 0 and WIT 30 groups. The amount of remaining RBCs was similar between all WIT 0 groups; whereas in the WIT 30 groups the amount was higher in kidneys washed out using HTK compared with PS. Histological evaluation showed less tissue injury among PS-washed kidneys compared with UW or HTK. CONCLUSION: Overall, kidneys washed-out with PS showed better preservation of structural integrity after 24 hours, CS compared with either UW or HTK. Washout of warm ischemically damaged kidneys was more effective using PS compared with HTK. PMID- 20005344 TI - Remifentanil, isoflurane, and preconditioning attenuate renal ischemia/reperfusion injury in rats. AB - BACKGROUND: The purpose of this investigation was to examine the effect of isoflurane, remifentanil, and preconditioning in renal ischemia/reperfusion injury (IRI). METHODS: All 52 male Wistar rats were anesthetized with isoflurane, intubated and mechanically ventilated. The animals were randomly divided into: S group (sham; n = 11) that underwent only right nephrectomy; as well as the I group of right nephrectomy and ischemia for 45 minutes by clamping of left renal artery. (n = 11); the IP (n = 9), the R (n = 10), and the RP (n = 11) groups. In addition, the R and RP animals received remifentanil (2 microg.kg(-1).min(-1)) during the entire experiment. The IP and RP group underwent ischemic preconditioning (IPC = three cycles of 5 minutes). Serum creatinine values were determined before and after IRI, as well as 24 hours later. In addition to an Histological study, cells from the left kidney were evaluated for apoptosis by flow cytometry (FCM). RESULTS: The Creatinine value of 0.8 +/- 0.2 mg/dl in the S group was significantly lower at 24 hours than the I 3.9 +/- 1.5 mg/dl; IP 2.6 +/ 1.7 mg/dl; R 3.3 +/- 2.8 mg/dl; or RP 1.8 +/- 0.5 mg/dl groups. The RP group value was significantly lower than those of the I, IP, and R groups (p < 0.05). The S group showed less proximal tubular cell damage than the I, IP, R, and RP groups (p < 0.05). The percentages of apoptotic cells (FITC(+)/PI(-)) were: S group = 11.6 +/- 6.5; I = 16.7 +/- 7.3; IP = 37.0 +/- 28.4; R = 11.7 +/- 6.6, and RP = 8.8 +/- 1.5. The difference between the IP vs RP group was significant. Similar percentages of necrotic cells (FITC(+)/PI(+)) and intact cells (FITC( )/PI(-)) were observed among the groups. CONCLUSIONS: Ischemic preconditioning showed no protective effect in the isoflurane group (IP) but when isoflurane was administered associated with remifentanil (RP), there was a beneficial effect on the kidney, as demonstrated by flow cytometry and serum creatinine values. PMID- 20005345 TI - Comparison of two models for evaluation histopathology of experimental renal ischemia. AB - Renal ischemia/reperfusion (I/R) injury is one of the frequent causes of acute renal failure (ARF) due to the complex, interrelated sequence of events, that result in damage to and death of kidney cells. Cells of the proximal tubular epithelium are especially susceptible to I/R injury, leading to acute tubular necrosis, which plays a pivotal role in the pathogenesis of ARF. Several models have been explicated to assess morphological changes, including those of Jabonski et al. and Goujon et al. We compared the 2 models for histopathological evaluation of 30- or 120-minute periods of renal ischemia followed by 24-hour reperfusion in rats. Several changes were observed after application of the 2 models: proximal tubular cell necrosis, loss of brush border, vacuolization, denudation of tubular basement membrane as a consequence of flattening of basal cells, and presence of intratubular exfoliated cells in the lumen of proximal convoluted tubules at various stages of degeneration (karyorexis, kariopyknosis and karyolysis). Evaluating tubular lesions after 2 periods of experimental ischemia with light microscopy allowed us to conclude that the Goujon classification better characterized the main changes in cortical renal tubules after ischemia. PMID- 20005346 TI - Influence on energy kinetics and histology of different preservation solutions seen during cold ischemia in the liver. AB - BACKGROUND AND PURPOSE: Cold flush preservation prolongs tissue viability during ischemia. However, there is little understanding of the effects of various preservation fluids on events during this period. A study of cold ischemia in rat livers was undertaken to compare biochemical and histological changes over time, using three preservation solutions: University of Wisconsin (UW), histidine tryptophan-ketoglutarate (HTK), and Leeds solution (LS) under development at our institution. Leeds solution is a phosphate-based sucrose solution that like UW contains the impermeant lactobionate and the metabolite allopurinol (1,5-dihydro 4H-pyrazolo[3,4-d]pyrimidin-4-one) which acts as a competitive inhibitor of xanthine oxidase, stopping the breakdown of hypoxanthine to xanthine by oxidizing it to alloxanthine, inhibiting both the conversion of hypoxanthine to xanthine and the conversion of xanthine to uric acid. MATERIALS AND METHODS: At various time points, samples were analyzed for adenosine triphospate (ATP) and metabolites by high-performance liquid chromatography as well as for histological changes. RESULTS: In all livers, ATP, ADP, and AMP degraded over 4 hours. In UW and LS groups, degradation beyond hypoxanthine was halted, and it continued in the HTK group. This blockade led to a significant reduction in the accumulation of xanthine and uric acid. Histological analysis showed protected architecture and maintenance of reticulin scaffolds in the UW and LS groups, whereas tissue breakdown was seen from earlier time points in the HTK group. Additionally, throughout ischemia, signs of pathological injury were more pronounced with UW- than with LS-preserved tissue. CONCLUSIONS: These results implied that cold ischemia in the liver is characterized by dynamic biochemical changes coincident with pathological injury which are initiated from the time of organ perfusion and influenced by the choice of the perfusion fluid. Allopurinol in UW and LS appears to be critical. We hypothesized that it may also affect the degree of subsequent reperfusion injury. The data supported the assertion that LS offerred improved preservation over UW, adding to the impetus to shorten ischemic times in clinical transplantation. PMID- 20005347 TI - Vitamin E succinate enhances steatotic liver energy status and prevents oxidative damage following ischemia/reperfusion. AB - We have previously shown that treatment of steatotic livers with vitamin E succinate decreases liver injury and increases survival after ischemia/reperfusion (I/R). It is now understood that compromised energy status is associated with increased injury following liver ischemia in the setting of hepatic steatosis at least partially as a result of increased reactive oxygen species (ROS) and induction of mitochondrial uncoupling protein-2 (UCP2). Given the association between ROS, mitochondrial function, and UCP2, it was our goal to determine whether the protective effects of vitamin E succinate were associated with decreased ROS injury, down-regulation of UCP2, or improvement of ATP levels following I/R. To test this, leptin deficient (ob/ob) mice with steatotic livers that had received other 50 IU of vitamin E succinate supplement per day or control chow for 7 days were subjected to total hepatic ischemia (15 minutes) followed by reperfusion. We measured liver expressions of ATP, glutathione (GSH), and UCP2 as well as mitochondrial DNA damage. Vitamin E treatment decreased hepatic UCP2 expression and increased ATP and GSH levels prior to I/R. These levels were maintained at 1 hour after I/R. At 24 hours, while hepatic UCP2 expression, ATP, and GSH levels were similar to those of mice not receiving vitamin E, mitochondrial DNA damage was blocked. These results revealed that vitamin E succinate decreased hepatic UCP2 expression, reduced oxidative stress, and improved mitochondrial function in mice with steatotic livers before and after I/R, identifying mechanisms of protection in this setting. PMID- 20005348 TI - Clotrimazole protects the liver against normothermic ischemia-reperfusion injury in rats. AB - OBJECTIVE: To investigate the possible antiapoptotic prosurvival role of the pregnane X receptor (PXR) in hepatic ischemia-reperfusion injury in rats using clotrimazole (CTZ), a strong PXR transactivator. MATERIALS AND METHODS: Male Sprague-Dawley rats were divided into 3 groups of 6 each: sham-treated, control, and CTZ-treated animals. Control and CTZ-treated animals were subjected to 30 minutes of normothermic ischemia of the whole liver followed by 6 hours of reperfusion. The animals were then killed, and the liver was excised and blood samples collected. RESULTS: Clotrimazole induced a significant increase in expression of the CYP3A gene, indicating PXR transactivation, whereas expression of the antiapoptotic Bcl-xL gene was not increased. Serum concentrations of aspartate aminotransaminase and alanine aminotransaminase were lower in CTZ treated animals than in control animals (difference not significant). Levels of poly(adenosine diphosphate-ribose) polymerase, a caspase-3 substrate, remained significantly higher in the CTZ-treated group compared with controls (P < .05). Clotrimazole increased the expression of phospho-p 44/42 extracellular signal regulated kinase 1,2 (P < .05). The gene expression of the heat shock proteins 27, 70 and 90 was significantly lower in CTZ-treated animals than in controls (P < .05). CONCLUSION: Clotrimazole-mediated PXR transactivation protects the liver against ischemia-reperfusion apoptosis in rats. Phospho-p 44/42 extracellular signal-regulated kinase 1,2 is activated, whereas gene expression of heat shock proteins 27, 70, and 90 is downregulated by induction of PXR. PMID- 20005349 TI - Pioglitazone attenuates ischemia/reperfusion-induced liver injury in rats. AB - INTRODUCTION: Hepatic ischemia/reperfusion (I/R) injury leads to free radical generation and acute inflammatory responses that cause liver damage, an important problem for liver transplantation. Pioglitazone is known to protect I/R injury in various tissues; however, the mechanism of cytoprotection is not well understood. This study investigated the effects of pioglitazone administration in a warm hepatic I/R model on tumor necrosis factor (TNF)-alpha level, tissue injury, and antioxidant enzyme activity. MATERIALS AND METHODS: Eighty wistar strain rats were divided into 4 groups (n = 20): Group 1 sham hosts; Group 2 hepatic I/R; Group 3 hepatic I/R + pioglitazone (10 mg/kg); and Group 4 hepatic I/R + vehicle. Rat livers were subjected to 30 minutes of ischemia followed by 6 hours of reperfusion. After reperfusion rats were humanely killed to obtain liver tissue to study glutathione peroxidase (GPx), superoxide dysmutase (SOD), malondialdehyde (MDA) levels and for histopathologic assessment. TNF-alpha, aspartate aminotransferase (AST), and alanine aminotransferase (ALT) were measured in serum. RESULTS: Pioglitazone pretreatment significantly reduced liver enzyme content (ALT, 176.80 +/- 13.75 vs 235.28 +/- 31.92 and AST, 748.20 +/- 79.29 vs 944.85 +/- 101.87) and TNF-alpha level (9:8.60 +/- 8.67 vs 138.28 +/- 9.99) after I/R compared with the control group. MDA level (3.02 +/- 0.37 vs 4.36 +/- 0.38) and hepatocytic degeneration were reduced in the pioglitazone-treated group. GPx (2.40 +/- 0.25 vs 1.36 +/- 0.31) and SOD activity (2.22 +/- 0.30 vs 1.40 +/- 0.35) were significantly higher in the pioglitazone-treated group compared with the control group. CONCLUSION: The present study showed that pioglitazone administration improved hepatic I/R injury that was associated with enhanced antioxidant enzyme activities and suppression of TNF-alpha, ALT, and AST levels. Because peroxisome proliferator-activated receptor-gamma agonists are widely used to treat diabetic patients, it may be relatively easy to expand their clinical indication. However, further investigations will be required to delineate protective mechanisms by which pioglitazone attenuates hepatic tissue injury after I/R. PMID- 20005350 TI - Effects of the antioxidants lycium barbarum and ascorbic acid on reperfusion liver injury in rats. AB - OBJECTIVE: Ischemia/reperfusion (I/R) of the rat liver can induce liver injury through mechanisms involving oxidative and nitrosative stresses. In this study we examined the effects of antioxidants Lycium barbarum (LB) and ascorbic acid on I/R-induced liver injury in rats. METHODS: Liver ischemia was induced by clamping the common hepatic artery and portal vein of rats for 40 minutes. Thereafter, flow was restored with reperfusion for 90 minutes. Blood samples collected before ischemia and after reperfusion were analyzed for alanine transaminase (ALT), lactic dehydrogenase (LDH), hydroxyl radical, and nitric oxide (NO) levels. Pharmacologic interventions included administration of ascorbic acid (100 mg/kg, i.p., 1 hour before I/R) or LB, an extract of Gogi berries: 600 mg in 100 mL of drinking water for 2 weeks prior to experimentation. RESULTS: This protocol resulted in elevation of blood concentrations of NO, hydroxyl radical, ALT, and LDH (P < .001) in the I/R-induced liver injury group. Ascorbic acid significantly attenuated the reperfusion liver injury by attenuating hydroxyl radical (P < .01) and NO (P < .05) release. The LB aggravated I/R-induced liver injury by increasing hydroxyl radical release with no effect on NO release. DISCUSSION AND CONCLUSIONS: This I/R protocol resulted in oxidative and nitrosative stress and liver injury. Ascorbic acid showed significant protective effects on reperfusion liver injury by attenuating hydroxyl radical and NO release. In contrast, LB aggravated liver injury by increasing hydroxyl radical release. PMID- 20005351 TI - Brain death and its influence on the lungs of the donor: how is it prevented? AB - The need for lung grafts is currently greater than ever. Unfortunately, the availability of grafts is insufficient for this demand. So we are forced to request organs for transplantation in the "waste bin." One possible solution to this problem may be the use of grafts from brain-dead patients. Sadly brain death is followed by devastating hemodynamic, inflammatory, and neurohumoral reactions in the potential donor which not only inflict direct damage, but also induce activation of the immune system which can cause rejection or even graft failure. Therefore, various groups have examined measures to prevent this outcome. In this review, we attempt to reconstruct the events that follow brain death, suggesting an algorithm to prevent a brain-dead patient's lungs from further damage. Finally, we are proposing potential measures of graft's protection of further investigation. PMID- 20005352 TI - Essential role for nuclear factor kappaB in ischemic preconditioning for cold ischemia-reperfusion injury of intestinal transplantation. AB - OBJECTIVE: The present study was designed to examine whether ischemic preconditioning could play a protective role on cold ischemia and reperfusion injury associated with intestinal transplantation in rats. METHODS: The 48 male Sprague Dawley (SD) rats were randomly assigned to 2 groups. Ischemic preconditioning was performed in experimental but not control rats by preserving the self donor small bowel in Ringer lactate solution at 4 degrees C for 3 hours (n = 6), 6 hours (n = 6), 12 hours (n = 6), or 18 hours (n = 6). One hour reperfusion was performed for every rat after orthotopic transplantation of donor small bowel. Small bowel samples were obtained for histological examination and immunohistochemistry analysis of nuclear factor kappaB (NF-kappaB) expression. RESULT: The small intestinal villus was arranged more regularly in experimental compared with control rats. Ischemia preconditioning also decreased edema in the muscule layer and increased Chiu score in experimental rats. Immunohistochemistry analysis revealed that ischemic preconditioning down-regulated the expression of NF-kappaB in the epithelia of experimental rats. CONCLUSION: Ischemic preconditioning improved intestinal transplantation in rats from cold ischemia and reperfusion injury by down-regulating the expression of NF-kappaB. PMID- 20005353 TI - Effect of donor ethnicity on kidney survival in different recipient pairs: an analysis of the OPTN/UNOS database. AB - BACKGROUND: Previous multivariate analysis performed between April 1, 1994, and December 31, 2000 from the Organ Procurement Transplant Network/United Network for Organ Sharing (OPTN/UNOS) database has shown that kidneys from black donors were associated with lower graft survival. We compared graft and patient survival of different kidney donor-to-recipient ethnic combinations to see if this result still holds on a recent cohort of US kidney transplants. METHODS: We included 72,495 recipients of deceased and living donor kidney alone transplants from 2001 to 2005. A multivariate Cox regression method was used to analyze the effect of donor-recipient ethnicity on graft and patient survival within 5 years of transplant, and to adjust for the effect of other donor, recipient, and transplant characteristics. Results are presented as hazard ratios (HR) with the 95% confidence limit (CL) and P values. RESULTS: Adjusted HRs of donor-recipient patient survival were: white to white (1); and white to black (1.22; P = .001). Graft survival HRs were black to black (1.40; P <.001); black to white (1.35; P <.001); black to Hispanic (0.87; P = .18); and black to Asian (0.69; P =.05). SUMMARY: Black donor kidneys are associated with significantly lower graft survival when transplanted into whites or blacks and are only associated with lower patient survival when these kidneys are transplanted into white recipients. The graft and patient survival rates for Asian and Latino/Hispanic recipients, however, were not affected by donor ethnicity. This analysis underscores the need for research to better understand the reasons for these disparities and how to improve the posttransplant graft survival rates of black kidney recipients. PMID- 20005354 TI - Potential differences in kidney allograft outcomes between ethnicities when converting to sirolimus base immunosuppression. AB - OBJECTIVE: The aim of this study was to determine whether ethnicity impacts graft outcomes in kidney transplant patients converted to sirolimus (SRL) and maintained on either calcineurin inhibitors (CI) or mycophenolate mofetil (MMF) with steroids. METHODS: This study analyzed kidney transplants converted to SRL and transplanted between July 1991 and April 2007. Patients were divided into 4 groups: group 1: African-Americans converted to SRL + CI; group 2: non-African Americans converted to SRL + CI; group 3: African-Americans converted to SRL + MMF; group 4: non-African-Americans converted to SRL + MMF. RESULTS: A total of 242 patients was included. Demographics, baseline immunosuppression, and reason for SRL conversion were similar among groups. Patients converted to SRL + CI regimens had significantly higher rates of acute rejection before SRL conversion, but equal rates after conversion. Development of proteinuria was similar across groups. African-American patients converted to SRL + MMF tended to have poorer outcomes compared with African-American patients converted to SRL + CI. Non African-American patients converted to SRL + MMF tended to have better graft outcomes compared with non-African-American patients converted to SRL + CI. CONCLUSIONS: African-Americans converted to SRL may benefit from continued CI, whereas non-African-Americans converted to SRL seem to have better outcomes with MMF. Further prospective studies are warranted to confirm these findings. PMID- 20005355 TI - Renal function outcomes in kidney transplant recipients after conversion to everolimus-based immunosuppression regimen with CNI reduction or elimination. AB - BACKGROUND: Chronic allograft nephropathy (CAN) is a major cause of progressive renal failure in kidney transplant recipients. Its etiology is multifactorial and can be due to immunologic or nonimmunologic conditions including calcineurin inhibitor (CNI) toxicity. OBJECTIVE: To evaluate the effect of conversion from CNIs to everolimus in kidney transplant recipients with CAN. PATIENTS AND METHODS: In this 12-month pilot study in renal transplant recipients with biopsy proved CAN, therapy was changed to an everolimus-based immunosuppression regimen. Cyclosporine or tacrolimus dosage was reduced by 80% (group 1, n = 10) or discontinued (group 2, n = 10). Mycophenolate mofetil or azathioprine were withdrawn in group 1, whereas both agents were maintained in group 2. All patients received prednisone. RESULTS: Twenty renal allograft recipients switched to an everolimus-based regimen, and patients were followed up for a mean (SD) of 12 (0.1) months. Baseline and end-of-study data were as follows: serum creatinine concentration, 1.27 (0.35) mg/dL vs 1.24 (0.4) mg/dL in group 1, and 1.27 mg/dL (0.36) vs 1.25 (0.3) mg/dL in group 2 (difference not significant); and estimated glomerular filtration rate, 72.4 (19.86) mL/min vs 76.26 (22.69) mL/min in group 1 (not significant), and 66.2 (12.95) mL/min vs 66.2 (13.73) mL/min in group 2 (not significant). One patient in group 1 experienced an acute rejection episode (Banff grade Ib), and 2 patients in group 1 and 1 patient in group 2 demonstrated borderline changes, all associated with everolimus blood concentration less than 3 ng/mL. CONCLUSIONS: Reduction or withdrawal of CNI and introduction of everolimus may be useful to slow the rate of loss of renal function in patients with CAN. PMID- 20005356 TI - Distinct cytokine patterns in different states of kidney allograft function. AB - Cytokines are crucial inflammatory mediators involved in the development of immune response leading to allograft rejection. We investigated the cytokine patterns in patients sera from cases of acute rejection episodes (ARE), chronic rejection (CR), and long-term stable courses (STABLE). The project included 20 patients with ARE, 20 with CR, and 15 with at least a 5-year stable course. Serum samples collected at the time of rejection diagnosis were cytometrically tested for concentrations of interleukin (IL) 2, IL-4, IL-6, IL-10, interferon (IFN) gamma, and tumor necrosis factor alpha. No significant differences between investigated groups were observed before transplantation (P > .05). Significant differences were observed among the groups in serum levels of IFN-gamma, IL-4, IL 6, and IL-10. Our data suggested that distinct serum cytokine patterns were present among various states of kidney allograft function. ARE was characterized by a mixed cytokine pattern with elevated IL-10 and IFN-gamma compared with the STABLE patients. The cytokine pattern in CR patients, in turn, was characterized by elevated levels of IL-4, IL-6, and IL-10 and decreased levels of IFN- gamma compared with both STABLE and ARE subjects. Our results suggested that the T(H)2 response may contribute to the initiation and/or maintenance of CR, because IL-4, IL-6, and IL-10 serve as growth and differentiation factors for B cells to increase antibody production. We also observed up-regulated production of IFN gamma and down-regulation of T(H)2 cytokines among patients with stable long- term graft function. PMID- 20005357 TI - Late acute kidney transplant rejection: clinicopathological correlates and response to corticosteroid therapy. AB - Acute rejection is a major cause of kidney allograft dysfunction. It is important to distinguish between cellular and antibody-mediated rejection to guide the treatment strategy. The management of acute antibody-mediated rejection includes aggressive therapy with plasmapheresis and intravenous immunoglobulin. C4d staining of peritubular capillaries has emerged as a valuable tool in identifying antibody-mediated rejection. Late acute rejection has a worse prognosis than early acute rejection. The clinical and pathological features of late acute kidney allograft rejection are not fully understood. We studied the clinicopathological correlates of late acute rejection in our patient population. During an 8-year period, all patients who had late acute rejection (6 months posttransplant) were identified. Patients with severe chronic changes and transplant glomerulopathy were excluded. Patients were divided into C4d+ and C4d- groups [corrected]. Histopathological features and treatment response were evaluated. Nine patients met inclusion criteria (4 C4d+, 5 C4d-). Maintenance therapy consisted of mycophenolate mofetil, calcineurin inhibitors, and low-dose prednisone. All patients received intravenous methlyprednisolone or high-dose oral prednisone as antirejection therapy. Seventy-five percent of patients in the C4d+ group and 80% of patients in the C4d- group had a clinical response to antirejection therapy. The majority of C4d+ patients with late acute rejection who were treated with corticosteroids alone responded to treatment. The study raises the possibility that a subset of C4d+ patients with acute rejection who do not have severe chronic changes might respond to corticosteroid therapy alone. PMID- 20005358 TI - Clinicopathological study of expression of lymphatic vessels in renal allograft biopsy after treatment for acute rejection. AB - BACKGROUND: Lymph vessel expression is related to inflammatory cell infiltration, around renal tubules in acute rejection episodes (ARE) of transplanted kidneys. However, there is little information on the lymph vessels after treatment of an ARE, particularly in relation to renal function and histological findings. PATIENTS AND METHODS: We investigated 13 cases of ARE diagnosed by kidney transplant biopsy performed from 1997 to 2005 within 3 years of transplantation. Treatment of the ARE lead to an improved serum creatinine level in all cases. There was neither an ABO-incompatible nor an acute humoral rejection case. Lymphatic vessels in re-biopsies were examined using immunohistochemical staining with D2-40 antibody that detected lymphatic endothelium. Re-biopsy cases in which the baseline creatinine had increased by more than 20% despite treatment were considered the severe group; the others, as the stable group. The relation between lymphatic vessel density (LVD) and renal function was examined using Banff scores. RESULTS: LVD was significantly higher in the severe than the stable group. The expression of lymph vessels versus the Banff score showed a direct relation: greater Banff scores showed higher expressions of lymph vessels. CONCLUSIONS: The expression of lymph vessels in renal allograft specimens after treatment of an ARE was related to deterioration of renal function and inflammatory cell invasion. We plan a further examination of the relationship between the expression of lymph vessels and long-term prognosis. PMID- 20005359 TI - Enteric-coated mycophenolate sodium reduces gastrointestinal symptoms in renal transplant patients. AB - A main cause for gastrointestinal (GI) complications in graft recipients is the routinely administered inosine monophosphate dehydrogenase inhibitor mycophenolic acid (MPA). MPA is available in two formulations, the prodrug mycophenolate mofetil (MMF) and the enteric-coated sodium salt (EC-MPS). Clinical results point to a better GI tolerability of EC-MPS as compared to MMF. We performed an open surveillance study in 397 organ graft recipients to investigate the clinical tolerability of EC-MPS in renal graft recipients who were converted from MMF to EC-MPS (maintenance) or who received EC-MPS as a new component of their immunsuppressive regimen (de novo). Physicians recorded GI symptoms (nausea, emesis, anorexia, diarrhea, abdominal cramps) at the start of EC-MPS treatment (visit 1) and at the next two visits in the clinic (visits 2 and 3); general tolerability (very good/good/moderate/poor) was assessed at visit 2 and 3. Two hundred seventy-five patients were on maintenance treatment with MMF and were converted to equimolar doses of EC-MPS, and 122 patients received EC-MPS de novo. The mean time since transplantation was 4.2 +/- 4.4 years. Median time until visit 2 was 28 days and until visit 3, 65 days. In 63.0% of patients, tolerability was rated as very good at visit 2 and in 64.7% at visit 3. Most patients who had suffered from GI complications during preceding MMF treatment reported improvement or total disappearance of their symptoms after conversion to EC-MPS. In conclusion, EC-MPS is a useful means to reduce GI complications in MPA treated patients. PMID- 20005360 TI - Low-dose dopamine in kidney transplantation. AB - BACKGROUND: The use of low-dose dopamine (LDD; 0.5-2.5 mug/kg/min) in kidney transplant recipients seeks to increase urine output, prevent arterial vasospasm, and reduce the incidence of acute tubular necrosis. The aim of this study was to evaluate the effect of LDD in the early postoperative period (12 hours) among kidney transplant recipients. METHODS: We studied all kidney transplant recipients admitted to the Intensive Care Unit (ICU) in the early postoperative period from January 2004 to December 2008. A total of 105 patients were retrospectively assigned to two groups: group A (n = 57) was treated with LDD and group B (n = 48), not treated with LDD. All patients otherwise received the same therapy. Blood sample analysis and kidney function were recorded at 0, 6, and 12 hours after admission. For each patient, we collected the following data: donor and transplant kidney features, recipient demographics, intraoperative events, hemodynamic and kidney function parameters in the ICU, and outcomes. Patients were followed for 6 months after ICU discharge. RESULTS: Hourly diuresis and kidney function parameters showed no significant difference between the groups. Significant differences between group A and group B were observed in heart rate (92.63 +/- 14.18 vs 82.87 +/- 13.5, respectively), hours of ICU length of stay (29.0 +/- 17.42 vs 20.43 +/- 7.35, respectively), and 6-month mortality rate (8.8% vs 0%, respectively; P < .05). CONCLUSION: LDD prescription in kidney transplantation neither improved kidney function during the postoperative period nor short-term outcomes, but was associated with an increased heart rate, ICU length of stay and 6-month mortality. PMID- 20005361 TI - Characteristics of long-term immunosuppressive therapy in Chinese pediatric renal transplant patients: a single-center experience. AB - We performed this study to investigate the trend and characteristics of various immunosuppressive regimens as well as their efficacy and safety for long-term survival of Chinese pediatric renal allograft recipients. METHODS: Thirty-four patients who underwent kidney transplantation between January 1985 and July 2002 had >/=5 years follow up. We retrospectively reviewed the baseline characteristics, patient and kidney survival rates, renal function, immunosuppressive regimens, drug levels, and adverse effects of immunosuppressive medications. RESULTS: The 1-, 3-, and 5-year recipient versus graft survival rates were 100% and 97.1%; 91.2% and 88.2%; 85.3% and 82.4%, respectively. The proportions of patients treated with cyclosporine- or tacrolimus-based immunosuppressive regimen at these times were 48.5%/51.5%; 60.0%/40.0%; and 53.6/46.4%. There were no significant differences in the dosages and drug levels after 1 year (P > .05). The proportions of azathioprine versus mycophenolate mofetil adjunctive therapy were 21.3/78.8%; 23.3%/70%; and 32.1%/60.7%, respectively. Forty percent of the surviving recipients developed complications, including hypertension, hyperlipidemia, gingival hyperplasia, hirsutism, liver dysfunction, herpes zoster, diabetes mellitus or cataracts. CONCLUSIONS: Cyclosporine or tacrolimus, plus mycophenolate mofetil or azathioprine, and prednisone triple therapies showed promising long-term results with similar efficacy and safety in pediatric renal recipients. Periodic drug level monitoring is required to facilitate individualization of immunosuppressive regimens. Drug doses and levels differed markedly from non-Chinese patients because of the ethnic discrepancy. PMID- 20005362 TI - Pilot study: association of traditional and genetic risk factors and new-onset diabetes mellitus following kidney transplantation. AB - INTRODUCTION: New-onset diabetes mellitus, which occurs after kidney transplant and type 2 diabetes mellitus (T2DM), shares common risk factors and antecedents in impaired insulin secretion and action. Several genetic polymorphisms have been shown to be associated with T2DM. We hypothesized that transplant recipients who carry risk alleles for T2DM are "tipped over" to develop diabetes mellitus in the posttransplant milieu. METHODS: We investigated the association of genetic and traditional risk factors present before transplantation and the development of new-onset diabetes mellitus after kidney transplantation (NODAT). Markers in 8 known T2DM-linked genes were genotyped using either the iPLEX assay or allelic discrimination (AD)-PCR in the study cohort testing for association with NODAT. We used univariate and multivariate logistic regression models for the association of pretransplant nongenetic and genetic variables with the development of NODAT. RESULTS: The study cohort included 91 kidney transplant recipients with at least 1 year posttransplant follow-up, including 22 who developed NODAT. We observed that increased age, family history of T2DM, pretransplant obesity, and triglyceridemia were associated with NODAT development. In addition, we observed positive trends, although statistically not significant, for association between T2DM-associated genes and NODAT. CONCLUSIONS: These findings demonstrated an increased NODAT risk among patient with a positive family history for T2DM, which, in conjunction with the observed positive predictive trends of known T2DM-associated genetic polymorphisms with NODAT, was suggestive of a genetic predisposition to NODAT. PMID- 20005363 TI - Statin therapy ameliorates renal allograft function. AB - BACKGROUND: Statins have proven ability as antilipidemic agents and benefit cardiovascular survival in transplant recipients. The pleiotropic effects of statins on renal function in renal allograft recipients are still undetermined. METHODS: Statin therapy was initiated according to guidelines for cardiovascular protection. Serum creatinine concentration and estimated glomerular filtration rate (eGFR) were analyzed before and after introduction of statins. The 73 patients who were retrospectively studied included those who were dialysis dependent. Mean changes in eGFR and lipid profile were compared before and after commencing statins using chi(2) tests. RESULTS: Mean serum creatinine concentration 18 months before starting statin therapy was 160.13 mumol/L, and 24 months after starting statin therapy was 172.22 mumol/L. Mean eGFR was 53.40 mL/min 18 months before starting statin therapy, and decreased to 49.43 mL/min after starting statin therapy. This represented a decline in renal function of 0.22 mL/min/mo over 18 months. The eGFR at 12 months after beginning statin therapy was 52.67 mL/min, and at 24 months was 49.06 mL/min. The rate of decline of eGFR after starting statin therapy was significantly lower: 0.02 mL/min/mo over 24 months (P < .001). Total cholesterol and low-density lipoprotein cholesterol concentrations were significantly decreased after starting statin therapy (P < .001). Four of 73 patients developed graft failure within 24 months. CONCLUSION: Statin therapy in our setting was associated with a lower rate of decline in renal function in renal allograft recipients within 2 years of starting treatment. PMID- 20005364 TI - Effects of cyclosporine-tacrolimus switching in posttransplantation hyperlipidemia on high-density lipoprotein 2/3, lipoprotein a1/b, and other lipid parameters. AB - OBJECTIVE: In renal transplant recipients, cyclosporine treatment appears to cause more frequent hyperlipidemia than tacrolimus usage. In this study, hyperlipidemic renal transplant recipients who use cyclosporine were investigated for changes in high-density lipoprotein (HDL)-2/3, apolipoprotein (Apo) A1/B, other lipid and biochemical parameters, and body mass index after prospective cyclosporine to tacrolimus switching. MATERIALS AND METHODS: Fifteen patients, including 9 females of overall mean age of 33.2 +/- 10.7 years and posttransplantation time of 78.06 +/- 42.93 months with a mean body mass index of 23.77 +/- 3.34 kg/m(2), were included if they were nondiabetic, hyperlipidemic, and had undergone renal transplantation between 1992 and 2000, using cyclosporine and candidates for a switch to tacrolimus due to hyperlipidemia. Before switching to tacrolimus and at 12 months of tacrolimus use we studied fasting blood samples for creatinine, uric acid, glucose, triglyceride, Apo A1, Apo B, low-density lipoprotein (LDL), HDL2, HDL3, and total cholesterol. RESULTS: There were no significant differences in creatinine, uric acid, glucose levels, or body mass index before tacrolimus versus 12 months thereafter. It was observed that tacrolimus significantly decreased triglyceride, Apo A1, Apo B, LDL, HDL, and total cholesterol levels (P < .001; P = .006; P = .01; P < .001; P = .03; P .05). CONCLUSION: Switching from cyclosporine to tacrolimus was associated with a more favorable cardiovascular risk profile by improving hyperlipidemia. PMID- 20005365 TI - Improved 24-hour blood pressure control with sirolimus versus calcineurin inhibitor based immunosuppression in renal transplant recipients. AB - INTRODUCTION: Calcineurin inhibitors (CNI) have brought dramatic improvements in early renal allograft survival. However, CNI are associated with posttransplant hypertension (PTHTN), a risk factor for mortality from cardiovascular disease and graft failure. Sirolimus (SRL) is emerging as an alternative to CNI. SRL effects on blood pressure (BP) in humans are unclear. We compared the prevalence of PTHTN among patients receiving SRL as maintenance immunosuppression with a group receiving CNI by using 24-hour ambulatory BP (AMBP). AMBP has been shown to predict cardiovascular events and progression of kidney disease better than casual office BP measurements in chronic kidney disease (CKD) patients. METHODS: Renal transplant recipients with office hypertension (defined as BP > 130/80 or on antihypertensive medications), receiving stable immunosuppression and displaying consistent serum creatinine values for > or =6 months were eligible. We enrolled the first 40 patients to consent. Office BP was measured twice using a BP-Tru machine. AMBP was then analyzed for systolic BP (SBP), diastolic BP (DBP), and nocturnal blood pressure fall (NF; "dipping"). Patients were placed in the SRL group (n = 18) and the CNI group (n = 20) based on their maintenance immunosuppressive protocol. Two patients were excluded because of incomplete data. All patients received mycophenolate mofetil, and 14/38, maintenance steroids. We collected, demographics as well as type and date of renal allograft, medications, comorbidities, CKD stage, proteinuria, and plasma creatinine at the time of study enrollment. RESULTS: Patients in the SRL group displayed lower 24 hour SBP than the CNI group (128.0 +/- 10.8 vs 137.7 +/- 14; P = .029). Nightime MAP, nightime SBP, and nighttime DBP were all lower in the SRL group. NF did not reach significance between the SRL and CNI groups (44% vs 15%; P = .074). Patient demographics and number of antihypertensive medications did not differ. CONCLUSION: The lower 24-hour SBP seen in the SRL group by AMBP may lead to improved cardiovascular and renal outcomes over time. Long-term patient follow-up will be needed to clarify the effect of the lower 24-hour SBP. PMID- 20005366 TI - Foxp3 staining in BK virus allograft nephropathy and comparison with acute cellular rejection. AB - BACKGROUND: Foxp3(+)CD4(+)CD25(+) regulatory T cells are involved in maintaining immunologic self-tolerance. These cells have been investigated in acute cellular rejection (ACR) of renal allografts. In this retrospective pathological study, we evaluated Foxp3(+) immunostaining in BK virus nephropathy (BKVN). In some circumstances, BKVN may be difficult to distinguish histologically from ACR. METHODS: Sequential sections were made of 30 allograft core biopsies and stained for hematorylin and eosin (H&E), C4d, cytomegalovirus (all negative), SV40, CD3, CD20, and Foxp3. Twelve biopsies were from diagnosed BKVN cases, 12 were from diagnosed ACR cases, and six showed neither BKVN nor ACR (controls). The 100x field of maximum cellular inflammation was located and marked on the H&E stain. The same area on the CD3, CD20, and Foxp3 slides was marked. Staining lymphocytes were counted under 400x magnification. Degree of BKVN was assessed according to the Drachenberg scale; degree of ACR was assessed by the Banff criteria. RESULTS: The range of Foxp3(+) staining (cells/mm(2)) was much larger in BKVN (0-270) compared to ACR (0-35). The mean difference did not reach statistical significance owing to a large degree of overlap between the two groups. In BKVN, the Foxp3(+) infiltrate correlated with the degree of CD3(+) infiltrate (P = .012), and median Foxp3(+) infiltrate increased with Drachenberg grade of BKVN. CD3(+) cell levels were not significantly different in BKVN versus ACR. CONCLUSIONS: BKVN cases with high levels of Foxp3(+) graft infiltrates may be manifesting an immune response different from that of ACR. Positive Foxp3 correlation with Drachenberg grade suggests a down-regulatory response. PMID- 20005367 TI - Preventing graft thrombosis after renal transplantation: a multicenter survey of clinical practice. AB - BACKGROUND: Renal allograft vascular thrombosis is a complication that often results in graft loss. Since there are no guidelines on immediate postoperative thromboprophylaxis, we performed a telephone survey of clinical practice in all renal transplantation centers in France. METHODS: Each center considered 4 cases relating to renal transplant candidates on dialysis with an increasing risk of thrombosis: Case 1: patient with no identified risk factors; Case 2: patient with an earlier incidence of deep vein thrombosis; Case 3: patient with ischemic heart disease on antiplatelet therapy; Case 4: patient with atrial fibrillation taking a vitamin K antagonist (VKA) with lupus nephritis syndrome. RESULTS: The treatments proposed by the centers (%) were: Case 1: No anticoagulation therapy (57.1%), calcium heparin at prophylactic doses (P-dose) (40%), or unfractionated heparin (UFH); (P-dose; 2.9%). Case 2: No anticoagulation therapy (34.3%), calcium heparin (P-dose; 51.4%), or UFH (P-dose; 5.7%). Case 3: (A) Interruption of aspirin (65.7%), and either no anticoagulation therapy (21.7%) or substitution of aspirin by calcium heparin (P-dose; 56.6%) or by UFH (P-dose; 8.7%). (B) No interruption of aspirin (34.3%), and either no additional prophylaxis (58.3%) or calcium heparin (P-dose; 33.3%). Case 4: Interruption of VKA (100%), and UFH at a curative dose (68.6%), UFH (P-dose; 14.3%), or calcium heparin (P-dose; 11.4%). CONCLUSIONS: Practices varied widely in the absence of studies of sufficiently high power. There is a need for a preoperative classification of thrombotic and hemorrhagic risk among renal transplant candidates and for consensus guidelines. PMID- 20005368 TI - Renal cell carcinoma of native kidney after renal transplantation: clinical relevance of early detection. AB - BACKGROUND: Life expectancy after transplantation has improved, and cancer may soon be the leading cause of late death after transplantation. The guidelines of the American and European societies of nephrology and urology have not yet established the optimal frequency for screening for renal cell carcinoma (RCC) of native kidneys in patients who have undergone renal transplantation. OBJECTIVE: To evaluate the prevalence, prognosis, and risk factors of RCC in a series of patients followed up for 16 years in our transplantation unit. MATERIALS AND METHODS: Our study is a follow-up observational cohort study conducted in 694 consecutive renal transplant recipients admitted to our institution from July 1991 through July 2007. At our institution, ultrasound studies of the native kidneys were performed every 6 months after renal transplantation. RESULTS: In the patient cohort studied, 10 patients developed a renal tumor (1.6% incidence). Three patients died of causes other than recurrence of RCC. Seven patients are alive with no evidence of RCC recurrence or metastatic disease after a mean (range) follow-up of 41 (12-96) months. Acquired cystic kidney disease and dialysis duration were positively associated with development of RCC. CONCLUSIONS: The incidence of RCC in the literature varies between 0.3% and 4.8%. The variability depends on the timing of follow-up, with a higher incidence in prospective studies with strict follow-up. We advise ultrasound studies performed by specialized physicians every 6 months after transplantation. More detailed guidelines designed by the major international transplantation societies are necessary. PMID- 20005369 TI - Postimplantation syndrome in a kidney trasplant patient: fever is not always synonimous with infection. AB - Fever in an immunocompromised patient like the transplanted subject is usually due to an overt or occult infection. Clinicians must make important decisions to find the cause of fever, and also concerning the timing and adequacy of empiric antibiotic therapy. However, occasionally, fever is not due to an infectious cause. A correct identification of its causes may avoid a needlessly prolonged course of antibiotics. Herein we have reported a kidney transplant patient with a febrile syndrome, which appeared after endovascular aortoiliac stent-graft placement, that was finally attributed to a noninfectious cause like postimplantation syndrome. PMID- 20005370 TI - Nucleosome serum levels in acute hepatic failure and MARS treatment. AB - Serum nucleosomes have been suggested to be markers for cell death and apoptosis. Increased hepatocyte apoptosis can be demonstrated in acute liver failure (ALF) as well as acute-on-chronic liver failure (ACLF). We investigated the relevance of nucleosomes in the setting of acute hepatic failure. Further, we studied the effects of the molecular adsorbent recirculating system (MARS) on this marker of cell death. We measured serum nucleosome concentrations with ELISA in 12 patients with ACLF and 7 patients suffering from ALF, with 14 patients experiencing stable chronic hepatic failure (CHF) as controls. In a subset of 8 ACLF and ALF patients treated with MARS, nucleosomes were determined immediately before and after the first MARS session. Baseline nucleosome serum concentrations were significantly increased in ACLF and ALF patients as compared with CHF patients (P = .0161 and P = .0037, respectively). There was no significant difference between the ALF and ACLF groups. Moreover, serum nucleosome levels did not change significantly during MARS treatment in ALF and ACLF patients. Serum nucleosome levels therefore may be useful to discern acute from chronic hepatic failure or to monitor the course and the severity of the disease. Our results, however, warrant further larger clinical studies regarding the clearance of nucleosome in artificial liver assist devices and to assess their role in acute hepatic failure. PMID- 20005371 TI - Vascular immunohistochemical markers: contributions to hepatocellular nodule diagnosis in explanted livers. AB - INTRODUCTION: The process of hepatic carcinogenesis involves a progression including large regenerative nodules, to dysplastic nodules, and finally to hepatocellular carcinoma. Angiogenesis is fundamental to the development of malignant tumors. Changes in sinusoidal capillarization and isolated arteries occur early in hepatic carcinogenesis. However, sometimes differentiation of hepatocellular nodules can be difficult for the general pathologist. The aim of this study was to evaluate angiogenesis by immunohistochemistry using CD34 and HHF35 antibodies for differential diagnosis of large regenerative nodules versus dysplastic nodules versus hepatocellular carcinoma using explanted cirrhotic livers. METHODS: Seventy-nine nodules obtained from 29 explanted cirrhotic livers were classified according to the International Working Party as follows: 17 large regenerative, 23 low-grade dysplastic, 23 high-grade dysplastic, and 16 hepatocellular carcinomas. These nodules were submitted to immunohistochemistry with antibodies to CD34 and HHF35 to analyze sinusoidal capillarization and arterialization, respectively. RESULTS: Semiquantitative analysis revealed that CD34 expression was >30% in dysplastic nodules and hepatocellular carcinoma; the staining in 93.8% of cases was diffuse, almost involving the entire sinusoidal lining in hepatocellular carcinoma. The number of isolated arteries was high in hepatocellular carcinoma (average, 4.369), which positively correlated with the other nodules (P < .005). CONCLUSION: Quantification of sinusoidal capillarization and isolated arteries in hepatocellular nodules, as detected with CD34 and HHF35 antibodies, respectively provided an important tool to differentiate dysplastic nodules from hepatocellular carcinoma. PMID- 20005372 TI - Living donor liver transplantation for congenital absence of the portal vein. AB - The congenital absence of the portal vein (CAPV) is a rare venous malformation in which mesenteric venous blood drains directly into the systemic circulation. Liver transplantation (OLT) may be indicated for patients with symptomatic CAPV refractory to medical treatment, especially due to hyperammonemia, portosystemic encephalopathy, hepatopulmonary syndrome, or hepatic tumors. Because portal hypertension and collateral circulation do not occur with CAPV, significant splanchnic congestion may occur when the portocaval shunt is totally clamped during portal vein (PV) reconstruction in OLT. This phenomenon results in severe bowel edema and hemodynamic instability, which negatively impact the patient's condition and postoperative recovery. We have successfully reconstructed the PV in living donor liver transplantation (LDLT) using a venous interposition graft, which was anastomosed end-to-side to the portocaval shunt by a partial side clamp, using a patent round ligament of the liver, which was anastomosed end-to end to the graft PV with preservation of both the portal and caval blood flows. Owing to the differences in anatomy among patients, at LDLT for CAPV liver transplant surgeons should seek to preserve both portal and caval blood flows. PMID- 20005373 TI - Comparison of stroke volume variations derived from radial and femoral arterial pressure waveforms during liver transplantation. AB - BACKGROUND: Stroke volume variation (SVV) is being increasingly used to predict fluid responsiveness. Since radial arterial pressure (RAP) and femoral arterial pressure (FAP) frequently showing discrepancies during liver transplantation (LT), we sought to investigate the effect of differing arterial waveforms on SVV and cardiac output (CO) derived from the Vigileo device, by comparing SVV and CO values derived from RAP (SVV(RAP), CO(RAP)) and FAP (SVV(FAP), CO(FAP)) during LT. METHODS: The linear associations and agreements between SVV(RAP) and SVV(FAP) and between CO(RAP) and CO(FAP) were assessed during LT. Hemodynamic variables were measured at nine predefined time points in all 32 recipients, resulting in 288 data pairs. RESULTS: Correlations were observed between SVV(RAP) and SVV(FAP) (r = .961) and between CO(RAP) and CO(FAP) (r = .848) at all time points. These correlations between SVV(RAP) and SVV(FAP) (r = .923) and between CO(RAP) and CO(FAP) (r = .902) existed even during the period when mean RAP and FAP values differed (10 minutes after reperfusion). Bland-Altman analysis for SVV(RAP) versus SVV(FAP) and for CO(RAP) versus CO(FAP) showed weak biases (-0.2% and -0.5 L/min) and reasonable limits of agreement (-2.2 to 1.8% and -1.9 to 0.9 L/min). The percentage errors for SVV and CO values were 27.0% and 22.2%. CONCLUSIONS: There was no significant difference between SVV(RAP) and SVV(FAP) when measured using the Vigileo device during LT. This finding indicated that SVV obtained using the Vigileo device offered relatively consistent information regardless of the catheterization site. PMID- 20005374 TI - A significant expansion of CD8+ CD28- T-suppressor cells in adult-to-adult living donor liver transplant recipients. AB - BACKGROUND: The appearance of human regulatory CD8(+) CD28(-) T-suppressor (Ts) cells has been associated with a reduced need for maintenance immunosuppression in cadaveric heart- kidney transplant recipients and pediatric liver-intestine transplant recipients. However, few data are available in adult-to-adult living donor liver transplantation (A-A LDLT). MATERIALS AND METHODS: To study the population of CD8(+) CD28(-) Ts cells in A-A LDLT, we performed flow cytometry on whole blood specimens obtained from 20 transplant recipients, 18 end-stage liver disease patients, and 20 normal controls. Meanwhile, we measured the trough levels of immunosuppressants and monitored graft function in transplant recipients. We retrospectively reviewed the clinical data of the 20 recipients. RESULTS: A significant expansion of CD8(+) CD28(-) Ts cells was observed among recipients of A-A LDLT as compared with a disease control group (P = .000) or healthy individuals (P = .000). All recipients were free of acute cellular rejection episodes. During the follow-up period, no grafts were lost due to acute or chronic rejection. CONCLUSION: Expansion of CD8(+) CD28(-) Ts cells in A-A LDLT seemed to be associated with a decreased occurrence of acute or chronic rejection and sustained good graft function. Based on our low dosages of immunosuppressants for recipients of A-A LDLT, we suggest that this strategy may promote expansion of CD8(+) CD28(-) Ts cells, which can conversely maintain the low immunosuppressant dosages. PMID- 20005375 TI - Nutrition support with glutamine dipeptide in patients undergoing liver transplantation. AB - AIM: The effect of total parenteral nutrition (TPN) support supplemented with alanyl-glutamine (Ala-Gln) dipeptide was investigated in a randomized, controlled clinical trial. METHODS: Sixty-five patients with the diagnosis of end-stage liver disease or hepatic cellular carcinoma admitted for orthotopic liver transplantation were randomly divided into 3 groups: diet group (n = 21), TPN group (n = 22), and Gln group (n = 22). Patients in the TPN and Gln groups received isocaloric and isonitrogenous TPN for 7 days. Venous heparin blood samples were obtained for assay on days 2 and 9 after surgery; we performed routine pathologic tests. RESULTS: Compared with the results on day 9 in the TPN group, there was a significant increase in the prognostic nutrition index and in prealbumin among the Gln group. Aspartate aminotransferase improved significantly by Gln treatment compared with traditional TPN support (P < .05). The pathologic results also showed Gln supplementation to reduce hepatic cell injury. A significant decrease in postoperative hospital stay was observed in the Gln group. CONCLUSIONS: Posttransplant TPN support greatly improved protein metabolism and nutritional state of patients. TPN with Ala-Gln helped to improve synthetic function and to reduce the injury to a transplanted liver. PMID- 20005376 TI - Increase of modified retrograde to antegrade flow ratio on doppler ultrasounds of the hepatic vein indicating tricuspid regurgitation during follow-up of liver transplantation: correlation with echocardiographic results. AB - The purpose of this study was to explore the best parameter of hepatic vein (HV) Doppler ultrasounds (DUS) that correlated with echocardiographic findings of and particularly the optimal cutoff value for tricuspid regurgitation (TR) following liver transplantation (LT). Thirty-six patients underwent echocardiography and DUS after LT from January 2006 to July 2007. Echocardiographic records were searched for TR grade and peak velocity of TR flow. The HV DUS parameters included peak velocity of retrograde flow (R), peak velocity of antegrade flow (A), the difference between R and A (R-A), the ratio of R to A (R/A ratio), and a modified R/A ratio, namely, the product of the R/A ratio and the R/A duration ratio. Correlation tests and receiver-operator characteristic analyses explored their interrelations and to obtained cutoff values to diagnose moderate and severe TR. TR grade best correlated with the modified R/A ratio (rho = 0.585), followed by the R/A ratio (rho = 0.503) and R (rho = 0.455). The modified R/A ratio was the most accurate parameter for the diagnosis of moderate and severe TR (Az = 0.825 and 0.895, respectively); its cutoff value was > or =0.11 for moderate TR (sensitivity and specificity both 77.78%) and 0.13 for severe TR (sensitivity, 100%; specificity, 81.2%). The modified R/A ratio best correlated with echocardiographic results of TR, although the strength of correlation was only moderate. Additionally, the modified R/A ratio was an accurate DUS parameter to diagnose moderate and severe TR among patients following LT. PMID- 20005377 TI - Ligation of left renal vein as a salvage procedure for splenorenal shunt after living donor liver transplantation: a case report. AB - We describe a case of recovered portal flow by ligation of the left renal vein (LRV) as a salvage procedure for a spontaneous splenorenal shunt (SRS) occurring the next day after right liver living donor transplantation (LDLT). Doppler ultrasonography showed normal graft portal venous flow immediately after LDLT, but nearly total diversion of portal flow into the existing splenorenal shunt was observed on the next day. Portal flow normalized after ligation of the LRV by relaparotomy. The patient recovered fully without complication and was discharged on the 17th postoperative day. He remains well at 14 months after the operation, returning to his previous occupation. This case was neither associated with acute rejection nor with small-for-size graft, which may increase intrahepatic vascular resistance, causing portal flow steal through shunts. Even though patients with preoperative SRS show normal portal flow immediately after transplantation, close monitoring is necessary for a possible decrease or loss of portal flow. If portal flow becomes insufficient, ligation of LRV as a salvage procedure is an important option that can be considered even after transplantation. PMID- 20005378 TI - The benefits of interferon treatment in patients without sustained viral response after living donor liver transplantation for hepatitis C. AB - Although it has been recognized that interferon (IFN) treatment is crucial for recurrent hepatitis C after liver transplantation, its benefits have not been determined among patients without a sustained viral response (SVR). METHODS: Eighty patients who received IFN plus ribavirin treatment after living donor liver transplantation were grouped as follows: group I (n = 18) SVR; group II (n = 25) no-SVR but viral response [VR] positive; Group III (n = 13) no-VR but biochemical response [BR] positive; and group IV (n = 24) no-VR and no-BR. RESULTS: In groups II and III, not only the histological activity grade and fibrosis stage, but also the serum parameters including transaminases and type IV collagen were stable for 3 years after induction of IFN-based treatment. In group I, the activity grade and fibrosis stage significantly improved (P < .01). In group IV, the fibrosis stage significantly deteriorated (P < .01); the serum transaminases and type IV collagen were significantly higher than the other groups (P < .01). The mean duration of IFN treatment was significantly longer among group II (96 weeks) compared with the other cohorts (P < .05). The 5-year graft survival rate in groups II (91%) and III (100%) were comparable to those of group I (100%); group IV (62%) was significantly lower than the other groups (P < .05). CONCLUSION: IFN treatment was beneficial even among subjects with IFN dependent VR or BR, although they did not achieve SVR. PMID- 20005379 TI - Evaluation of Anti-HBs serum levels and pharmacokinetic profile after intravenous administration of Niuliva, a new hepatitis B immunoglobulin, following liver transplantation. AB - PURPOSE: We assessed whether trough anti-hepatitis B surface antigen (HBs) serum levels considered to be protective (>100 IU/L) were maintained in liver transplanted patients after 6 months of uninterrupted treatment with Niuliva, a new intravenous HBIg. METHODS: Twenty patients, aged 18-70 years, who had undergone liver transplantation due to HBV-related liver disease at least 1 year before inclusion were enrolled in a prospective, open-label, uncontrolled, multicenter clinical study. A fixed monthly dose of 5,000 IU of study medication was administered intravenously for 6 months. RESULTS: After the second infusion, all mean values of anti-HBs and 95% CIs were above the limit of 100 IU/L considered to be protective. The percentages of success ranged between 95% (95% CI 75.1%-99.9%) and 100% (95% CI 86.1%-100%). Mean values and 95% CI of in vivo recovery of anti-HBs at 15-30 minutes after each infusion showed overlaps between all intervals, which indicated that significant differences were not present in the recovery of post infusion anti-HBs in vivo. There were no recurrences of HBV infection during the study. There were no seroconversions in patients previously negative to hepatitis C virus or HIV. No serious adverse events related to the study medication were observed. CONCLUSIONS: Serum levels of anti-HBs after using Niuliva in patients who had undergone liver transplantation were protective in 95%-100% of cases after the study period. This new HBIg showed the expected pharmacokinetic profile and was well tolerated and safe. PMID- 20005380 TI - The new method of time-lag ligation for portosystemic shunt using coronary artery bypass graft occluder for adult living donor liver transplantation. AB - We performed a living donor liver transplantation (LDLT) for a 57-year-old man who had end-stage liver failure with portal hypertension and an inferior mesenteric vein-left testicular vein (IMV-LTV) shunt. At operation, we did not clamp the shunt but encircled it with a coronary artery bypass graft (CABG) occluder (Sumitomo Bakelite K.K., Japan), which was passed outside the body through the abdominal wall to time-lag ligation (TLL). On postoperative day (POD) 5, we observed decreased portal flow. We performed TLL of the shunt using the CABG occluder without re-laparotomy. The portal flow increased, while the portal vein pressure increased slightly. In LDLT, portosystemic shunt has been reported to be a cause of portal thrombus formation or graft liver atrophy due to decreased PV flow in the mid postoperative period. However, perioperative ligation of a portosystemic shunt may prevent regeneration of the grafted liver because of excessive portal hypertension. Therefore the technique of time-lag ligation of a portosystemic shunt using a CABG occluder may be a minimally invasive, useful method to achieve physiological liver graft regeneration. PMID- 20005381 TI - Mycophenolate mofetil versus enteric-coated mycophenolate sodium after simultaneous pancreas-kidney transplantation. AB - INTRODUCTION: Adverse gastrointestinal events are frequent after mycophenolate use. The objectives of the present study were to report the incidence of acute noninfectious diarrhea, to determine the risk factors, and to compare the severity of reactions between mycophenolate mofetil (MMF) and enteric-coated mycophenolate sodium (EC-MPS) after simultaneous pancreas kidney transplantation (SPKT). METHODS: We included 165 SPKT patients from December 2000 to May 2007. Uni- and multivariate analyses were performed, using acute noninfectious diarrhea as the dependent variable. P < .05 was considered significant. RESULTS: Mean age and duration of dialysis and of diabetes were 34.9 +/- 8.2 years, 27.3 +/- 18.3 months, and 21.9 +/- 16.2 years, respectively. Sixty-three percent used MMF, 36.4% used EC-MPS, and 0.6% used azathioprine. Multivariate analysis showed that the duration of diabetes (P = .049, confidence interval [CI] 1.0- 1.13) and MMF use (P = .013, 95% CI 0.2-0.82) were the main determinants of acute diarrhea after SPKT. MMF dose reduction (79.2% vs 62.3%, P = .024) and severity of diarrhea associated with orthostatic hypotension were more pronounced among MMF than EC-MPS patients (42.4% vs 15.1%, P = .001). There was no difference between MMF and EC-MPS after dose reduction in relation to the occurrence of acute kidney rejection (30.8% vs 26.7%, P = .53). CONCLUSIONS: Acute noninfectious diarrhea after SPKT was related to the duration of diabetes and to prescription of MMF. Preferential use of EC-MPS was associated with a lower necessity of dose reduction and less severe episodes of acute diarrhea compared with MMF, although dose reduction was equally associated with acute episodes of kidney rejection. PMID- 20005382 TI - One hundred pancreas transplants performed in a Brazilian institution. AB - After decades of controversy surrounding the therapeutic validity of pancreas transplantation, the procedure has become accepted as the preferred treatment for selected patients with type 1 diabetes mellitus. Between January 2001 and January 2008, 100 patients underwent pancreatic transplantation at our center: 88 simultaneous pancreas-kidney transplantation and 12 pancreas transplantations alone. Pancreas graft management of the exocrine drainage technique involved enteric drainage in 8 (all simultaneous pancreas-kidney) and the bladder in 92 cases. The recipient systemic venous system was used for the pancreas graft venous effluent in all cases. Our overall results have shown that the number of functioning pancreatic grafts was 64 of 100. Graft losses were: rejection (n = 8), venous thrombosis (n = 9), arterial thrombosis (n = 1), or surgical complications such as anastomotic leak (n = 3), perigraft infection (n = 10), pancreatitis of the graft (n = 5). Most cases of pancreatitis (80%) had preservation times exceeding 18 hours. Despite surgical and immunosuppressive complications, our impression was that pancreas transplantation was a highly effective therapy for diabetes mellitus. After 7 years of the program and 100 transplantations, we believe that there is a major role for transplantation in diabetes management. PMID- 20005383 TI - Availability of pancreatic allograft biopsies via a laparotomy. AB - OBJECTIVE: The aim of this study was to evaluate the availability of a pancreatic allograft biopsy via a laparotpmy. PATIENTS AND METHODS: From September 2004 to November 2007, 17 pancreas transplantations were performed: 15 simultaneous pancreas and kidney transplantations (SPK), 1 pancreas transplant alone (PTA), and one pancreas after kidney transplantation (PAK). Thirteen pancreatic allograft biopsies were obtained via an open laparotomy. This study evaluated the complications associated with this procedure, the rate of obtaining an adequate sample, and the relationship between biopsy-proven rejections and laboratory markers. In SPK cases we evaluated the synchronization between pancreas and kidney rejection. The pancreatic samples were diagnosed according to the Drachenberg classification. RESULTS: No complications resulted from the procedure. The rate of obtaining adequate samples was 84.6%. Pancreas rejection correlated with elevation of the laboratory markers in 71.4%. Simultaneous pancreas and kidney rejection occurred in 62.5%, only kidney in 25%, and only pancreas in 12.5%. CONCLUSION: A pancreas graft biopsy was absolutely imperative to improve the outcome in PTA, and even in SPK cases. A pancreatic allograft biopsy via a laparotomy was a safe, necessary and easy procedure to obtain an accurate diagnosis of rejection among pancreas transplantation patients. PMID- 20005384 TI - Validation of limited sampling strategy for estimation of mycophenolic acid exposure during the first year after heart transplantation. AB - BACKGROUND: Individualization of mycophenolate mofetil (MMF) dosing based on mycophenolic acid (MPA) therapeutic drug monitoring may minimize the risk of organ transplant rejection. The MPA area under the 12-hour concentration-time curve (MPA-AUC(0-12h)) is a more powerful predictor of rejection than are MPA trough levels. Measurement of MPA-AUC(0-12h,) however, is difficult and clinically impractical. The limited sampling strategy (LLS) has been proposed to overcome this problem. OBJECTIVE: To validate the predictive performance of MPA LSS algorithms previously published for heart transplant (HTx) recipients (initial group) when applied to a new independent group of 29 HTx recipients (validation group) during the first year after transplantation. PATIENTS AND METHODS: In a previous study, we established 2 algorithms using a LSS in HTx recipients: (1) 5.568 + 0.902 x C(1.25) + 2.022 x C(2) + 4.594 x C(6) and (2) 3.8 + 1.025 . C(1.25) + 1.819 x C(2) + 1.566 x C(4) + 3.479 x C(6). Agreement between abbreviated AUC and the full AUC(0-12h) was tested using the Bland-Altman method. The validation group was used to test and assess bias and precision. RESULTS: The 2 LSS algorithms used predicted the corresponding MPA-AUC(0-12h) with a mean bias of -4.85% and -3.6% and mean precision of 15.9% and 14%, respectively. CONCLUSIONS: The MPA-AUC(0-12h) obtained using the LSS may be useful to guide clinical management and dosing. This study prospectively validates 2 algorithms for calculation of MPA-AUC(0-12h) using an LSS calculated in HTx recipients. Bias and precision values suggest that our algorithms could be used for MPA therapeutic drug monitoring predictions in HTx recipients who share the same characteristics. PMID- 20005385 TI - Immunosuppression using the mammalian target of rapamycin (mTOR) inhibitor everolimus: pilot study shows significant cognitive and affective improvement. AB - Immunosuppression using calcineurin inhibitors (CNIs) is accompanied by neuropsychiatric side effects, which counteract longevity and quality of life benefits in 10% to 28% of patients. Following the availability of the mammalian target of rapamycin (mTOR) inhibitors, it became possible to replace CNI without increasing the risk of acute graft rejection. mTOR, a member of the phosphatidyl inositol 3' kinase family, is a downstream target of brain-derived neurotrophic factor, which has been implicated in the pathophysiology and treatment of several psychiatric disorders. Preclinical evidence has implicated the mTOR pathway in synaptic plasticity and fear memory consolidation and reconsolidation. METHODS: In the present study we prospectively evaluated the psychiatric outcomes of CNI free immunosuppression in adult maintenance heart transplant recipients (n = 9; age: 66.1 +/- 6.1) using the Wechsler Memory Scale-Revised (WMS-R), Symptom Checklist-90-Revised (SCL-90-R), Beck Depression Inventory (BDI), Trail Making Tests A and B, Digit Span (DS), and Hamilton Depression Scale (HAMD). RESULTS: Four weeks after switching to CNI-free immunosuppression using everolimus, BDI (Z = -1.14; P = .048), Trail Making tests A and B (Z = -2.52; P = .012), WMS-R (Z = 2.37; P = .018), and SCL-90-R (Z = -2.37; P = .018) were all significantly improved while DS (Z = -1.18; P = .236) and HAMD (Z = -0.595; P = .552) remained unchanged. CONCLUSION: This report describes favorable psychiatric outcome variables using everolimus in maintenance heart transplant recipients. CNI-free immunosuppression with everolimus might provide significant improvement in memory, concentration, and overall psychiatric symptoms among heart transplant recipients. PMID- 20005386 TI - Heart transplantation in a low-organ-donation environment: a single center experience. AB - OBJECTIVE: Heart transplantation is the "gold standard" for treating patients in end-stage heart failure who satisfy strict selection criteria. However, infrequent transplant performance, eg, less than nine per year, may be associated with suboptimal results. METHODS: We reviewed our 13-year clinical experience (1996-2008) with 73 orthotopic heart transplants performed under strict selection criteria and followed closely thereafter at the only accredited center in Greece, a country with an annual rate of only seven donors per million population. RESULTS: Low perioperative (5.47%) and long-term (7.5%) mortality rates were responsible for a 94% survival rate in the first year, 92% at five years, and 70% at ten years-similar to those reported worldwide-along with excellent functional recovery. CONCLUSION: Strict recipient and donor selection criteria, combined with a rigorous multidisciplinary follow-up, yield excellent results despite the existing shortage of available grafts. PMID- 20005387 TI - Improved, low-cost methods for pancreatic islet purification in rats. AB - OBJECTIVE: Density gradient separation of islets from exocrine tissue is usually performed using Ficoll. However, this reagent significantly increases the cost of isolation. The aim of the present study was to investigate the effects on islet preparations of purification methods using Lymphoprep and Iodixanol (OptiPrep) density gradients. METHODS: Pancreata were procured from 46 Wistar rats, loaded with collagenase V (Sigma), and mechanically dissociated using standard procedures. After the digestion phase, the islets purified by 3 methods-Ficoll, Lymphoprep, and Iodixanol (OptiPrep)-were assessed for yields, purity, morphology, and in vitro function. RESULT: We expressed the yields as islet equivalents (IEQ, diameter standardizing to 150 microm), showing no significant differences. Compared with the Ficoll group, the purity was significantly higher in the Lymphoprep (P = .005) and Iodixanol (OptiPrep) groups (P = .011). While the viability was >90% in all 3 groups, the viability in the Lymphoprep Group and OptiPrep groups was significantly higher than that of the Ficoll group (P < .001). In vitro islet function did not differ among the 3 experimental groups. CONCLUSION: Lymphoprep and Iodixanol were as effective as Ficoll in terms of islet yield and in vitro function. High-purity and high-viability islet cells were obtained using improved Lymphoprep-based or Iodixanol (OptiPrep)-based density gradient methods, potential low-cost substitutes for Ficoll. PMID- 20005388 TI - Improved islet survival and funtion with rat endothelial cells in vitro co culture. AB - OBJECTIVE: Pancreatic islet transplantation is an emerging therapy for type 1 diabetes. To preserve its function, transplanted islets must be revascularized because arterial and venous connections are disrupted during islet isolation. The current paradigm is that islet revascularization originates from the transplant recipient. This study was designed to test whether the function of isolated islets can be retained by co-culture with thoracic aorta endothelial cells in vitro. METHODS: Sprague-Dawley rats were used in this study. The endothelial cells (ECs) were isolated from the thoracic aorta. The viability of the isolated islets was assessed by acridine orange/propidium iodide (AO/PI) double staining. The islets were either placed in standard cultures (group A) or in co-cultures with ECs (group B). Islet viablity was assessed by an insulin release assay. RESULTS: The islets in group B exhibited normal morphology with >90% staining positive as detected by AO/PI with 7 days. Insulin release assays showed a significantly higher simulation index (SI) in group B compared with group A (P < .05) except on the first day. CONCLUSION: This study suggested that co-cultrue of freshly isolated rat islets with ECs improves postculture survival and islet function in vitro. PMID- 20005389 TI - Novel sulfated glucomannan-barium-alginate microcapsules in islet transplantation: significantly decreased the secretion of monocyte chemotactic protein 1 and improved the activity of islet in rats. AB - The sulfated glucomannan can be used to filter the heparin-binding properties of cytokines. In this study, novel sulfated glucomannan-barium-alginate (SGA) microcapsules were prepared to encapsulate islets with barium-alginate (ABa) and calcium alginate-poly-l-lysine (APA) microcapsules as controls. SD rat islets were purified as donor cells to Lewis rats that had been treated with streptozotocin. Intraperitoneal transplantation was performed with about 3000 islet equivalent (IEQ) rat. At week three after transplantation, the concentrations of monocyte chemotactic protein-1 (MCP-1), interleukin (IL)-1 beta, interferon (IFN)-gamma, and tumor necrosis factor (TNF)-alpha in intraperitoneal fluid were determined using ELISA. At week 8, the islet cell mass in the abdominal microcapsules was excised to test insulin release. The EB-FDA fluorescence staining method was used to observe the functional activity of the islet cells. Compared with ABa and APA microcapsules, SGA microcapsules showed significantly decreased MCP-1 secretion by beta-cells. Also, the concentrations of cytokines IL-1beta, IFN-gamma, and TNF-alpha were decreased significantly. The activity of the transplanted islets was significantly improved in SGA microcapsules, which shielded against cytokines better than ABa or APA microcapsules and may serve as novel method. PMID- 20005390 TI - Effect of low-level laser irradiation on in vitro function of pancreatic islets. AB - INTRODUCTION: Although islet isolation and transplantation techniques have improved extensively in recent years, the loss of healthy functional islets is one of the major obstacles in this enterprise. A biostimulatory effect of low level laser irradiation has been proven on proliferation of some kinds of cells. The aim of this study was to evaluate the effect of low-level laser irradiation on the function of isolated rat pancreatic islets after 24 hours of preculture. METHODS: Pancreatic islets isolated from male rats (250 to 300 g) were cultured for 24 hours in RPMI 1640 media. Groups of islets then received different energy densities (1, 3, 5 joules/cm(2) or silent) at 2 wavelengths (810 nm and 630 nm) using laser devices. Insulin concentrations in buffer media were measured as indices of islet function. RESULTS: Irradiation of incubated islets with 830 nm low-level laser significantly increased insulin secretion after a glucose challenge test (P < .05). There was a significant increase in insulin secretion after irradiation with joules/cm(2) 630 nm energy density (P < .001). CONCLUSION: These findings suggest that low-level laser irradiations improved islet cell function before transplantation. PMID- 20005391 TI - Increased T-bet to GATA-3 ratio during acute allograft rejection in the rat lung. AB - Acute allograft rejection (AR) remains a major problem in solid organ transplantation. The pivotal mechanism hinges on alloantigen recognition by recipient T helper (T(h)) cells that differentiate into T(h)1 and T(h)2. This study investigated the association of mRNA levels of the transcription factors T box expressed in T cells and GATA-binding protein 3 with the development of T(h)1/T(h)2-directed immune responses. We investigated the expression of T-bet and GATA-3 mRNA levels and the protein levels of their marker cytokines interleukin (IL)-2 and IL-4 in orthotopically transplanted rat lungs during AR. We observed a nonsignificant increase in T-bet expression following allografting at days 3 and 5 but there was a significant reduction in GATA-3 expression on day 5 compared with controls. The ratio of T-bet to GATA-3 expression showed a trend to increase at day 3 following allografting reaching significance at 5 days. These changes were associated with a significant increase in the expression of IL 2 over IL-4 on days 3 and 5. This study suggests that T(h)1 responses play a major role during AR in the rat lung, and that this differentiation can be monitored by measuring mRNA levels of T-bet and GATA-3. PMID- 20005392 TI - Prolonged cardiac allograft survival in mouse model after complement depletion with Yunnan cobra venom factor. AB - BACKGROUND: Activation of the complement system is the leading mechanism that causes antibody-mediated acute rejection and hyperacute rejection after xenotransplantation. The major cause of acute rejection in allogeneic transplantation is the T cell-mediated specific immune response. We studied the effects of complement on acute rejection after cardiac allotransplantation using complement depletion with cobra venom factor (CVF) in the mouse. MATERIALS AND METHODS: The Balb/c-C57 mouse model of heterotopic cardiac allograft was used. The mice were divided into 2 groups, a control group and a CVF-treated group. After intravenous injection of CVF, the experimental group was observed for allograft survival time. Twelve mice from the control and experimental groups were sacrificed on days 3, 5, and 7 after the operation. The pathologic grade of acute rejection, deposition of C3 in tissue, extent of infiltration by CD4+ and CD8+ T cells, and expression of MHC-II, B7-1, and B7-2 were compared between the 2 groups. RESULTS: In the CVF-treated group, mean (SD) survival of the cardiac allograft was 26.2 (1.7) days, and in the control group was 8.4 (0.4) days (P < .01). Pathologic examination and immunohistochemistry demonstrated that the grade of acute rejection, deposition of C3 in tissue, extent of infiltration of CD4+ and CD8+ T cells, and expression of MHC-II, B7-1, and B7-2 were significantly decreased in the CVF-treated group. CONCLUSION: Depletion of complement in the serum with CVF inhibits acute cardiac allograft rejection in the mouse. PMID- 20005393 TI - Antirejection effect of Herba Asari extract in rats after cardiac allograft implantation. AB - OBJECTIVE: We studied the antirejection effect of asarinin, the extract of Herba Asari, in rats that underwent cardiac allograft implantation. METHODS: Rats received either olive oil (as a placebo control) or cyclosporine, with various amounts of asarinin or a combination of asarinin and cyclosporine daily through oral administration. 102 recipient SD rats were divided randomly into 7 groups: 1) group A, treated with 8 ml/kg olive oil per day; 2) group B, treated with 2.5 mg/kg cyclosporin A (CsA) each day; 3) group C, treated with 5 mg/kg cyclosporin A (CsA) each day; 4) group D1, treated with 12.5 mg/kg asarinin each day; 5) group D2, treated with 25 mg/kg asarinin each day; 6) group D3, treated with 50 mg/kg asarinin each day; 7) group E, treated with 12.5 mg/kg asarinin and 2.5 mg/kg CsA each day. Allograft survival was observed in each group of rats. On the seventh day posttransplantation; we analyzed weights, pathological lesions, and the levels of interleukin (IL)-2, interferon (IFN)-gamma, IL-4, and IL-10 in peripheral blood. RESULTS: The survival time of donor hearts in the asarinin groups was significantly prolonged (P < .01) when compared with the control group (7.8 +/- 0.7 days). The weights of rats increased in group C, group D2, group D3, and group E at 7 days after operation. Pathological lesions were significantly less severe. The levels of IL-2 and IFN-gamma decreased remarkably (P < .01), while those of IL-4 and IL-10 were not affected (P > .05) in the asarinin groups (D2 and D3). CONCLUSIONS: Asarinin decreased peripheral blood concentrations of IL-2 and IFN-gamma with prolongation of allograft heart survival. PMID- 20005394 TI - Expression of NKG2D and its ligand in mouse heart allografts may have a role in acute rejection. AB - BACKGROUND: Ligands for the natural killer cell-activating receptor NKG2D, such as retinoic acid early inducible (Rae-1), minor histocompatibility antigen H60 (mouse), and major histocompatibility complex class I chain-related (human) may be expressed by tissues in response to stress. Because NKG2D-ligand engagement may induce natural killer cell activation and provide T-cell costimulation, we examined whether this interaction between innate and adaptive immunity occurred during heart transplant rejection. METHODS: Hearts from BALB/c mice were heterotopically transplanted into C57BL/6 mice without immunosupression. Grafts were harvested at 1, 3, and 5 days after transplantation. Rae-1, H60, and NKG2D mRNA were analyzed by RT-PCR, and the proteins were detected by immunohistochemistry. RESULTS: Compared with no expression in naive BALB/c mice hearts, Rae-1 mRNA levels in heart allografts were detected from days three to five postoperative, H60 on day five, and NKG2D on day three but prominently on day five postoperative. Immunohistochemical assay showed that compared with rare expression in syngeneic cardiac grafts, there were significant protein expressions of Rae-1 and NKG2D in heart allografts from days three to five postoperative and of H60 on day 5 postoperative. CONCLUSION: This study reported significant mRNA and protein expression of Rae-1, H60, and NKG2D during acute cardiac allograft rejection. The simultaneous and significant expression of NKG2D and its ligands indicated that interactions with innate immunity may promote acute rejection. The results also suggested that Rae-1 and H60 may be new targets to amelioate this immune response. PMID- 20005395 TI - Addition of adipose-derived stem cells in cord blood cultures stimulates their pluripotent differentiation. AB - INTRODUCTION: Adipose tissue is recognized as an important source of postnatal mesenchymal stem cells for generative medicine applications. Moreover, cord blood stem cells have been shown to contain pluripotent stem cells called unrestricted somatic stem cells (USSCs). However, this population is rare and cannot be generated from every cord blood sample. In this study, we have presented a new method of co-culture of adipose-derived stem cells (ADPCs) and cord blood stem cells that results in pluripotent differentiation. MATERIALS AND METHODS: ADPCs were obtained from a piece of adipose tissue after treatment with 0.075% collagenase, which was subsequently inactivated with DMEM/10% FBS. The cellular pellet of centrifugation was plated at 5-7 x 10(6) cells/mL in T25 culture flasks in a low-glycose DMEM with 30% FCS. Cord blood stem cells were obtained by centrifugation following double-processing in the presence of 2% HES 200/0.5 and plated at 5-7 x 10(6) cells/mL in the same medium. To investigate the crucial role of ADPCs in pluripotent cord blood differentiation, we added a ADPCS as (1 x 10(4) cells/mL) to the cord blood cultures and analyzed the contribution of ADPCs using a microscope as well as with flow cytometry. RESULTS: After only 3 days, adherent cells (USSC colonies) of fibroblastic morphology were detected in all co cultured samples, whereas this was observed later or not at all in the non-co cultured samples. The greater density of colonies in the co-coltured samples was another point. Hematopoietic CD45 cells were no longer detected after the first passage. Pluripotent stem cells were obtained from all co-cultured samples that contained stem cells positive for CD29, CD44, CD49e, CD90, CD105, CD51 Stro, and C-kit antibodies but negative for CD34, CD45, CD133, and glycophorin A. CONCLUSION: Addition of ADPCs was crucial to generate pluripotent-derived stem cells from cord blood samples. This double culture may be a useful tool for a universal allogeneic stem cell source for tissue repair or regeneration. PMID- 20005397 TI - Bone marrow-derived mesenchymal stem cells inhibit acute rejection of rat liver allografts in association with regulatory T-cell expansion. AB - Bone marrow-derived mesenchymal stem cells (MSCs) exhibit immunosuppressive functions in vitro and in vivo. We investigated the immunoregulatory effects of rat MSCs in a model of allogeneic liver transplantation. Brown Norway rats received livers from inbred Lewis rats, and at designated intervals, infusions of MSCs derived from recipient, donor, or third-party rats. Allograft rejection and recipient survival rates were recorded. In particular, changes in circulating regulatory T cells (Tregs) were measured. After administration of MSCs derived from each of the 3 strains, allograft recipients demonstrated markedly longer survival compared with control animals. Histologic analysis revealed significant inhibition of allograft rejection. The MSCs induced generation of CD4+CD25+Foxp3+ Tregs. We concluded that MSCs inhibit acute rejection of allografts after liver transplantation, and propose that the immunoregulatory effects of MSCs are associated with expansion of Tregs. PMID- 20005396 TI - Hair stem cells for bladder regeneration in rats: preliminary results. AB - BACKGROUND: A variety of tissue engineering techniques are currently under development or investigation for bladder augmentation, but so far no approach is clearly superior. The aim of this study was to compare the suitability for cystoplasty augmentation in rats of in vivo implanted acellular bladder matrices (BAM) previously seeded with hair follicle stem cells and that of matrices implanted without the cells. MATERIALS AND METHODS: The rat hair follicle stem cell line was positive for CD34, p63, and Ki-67. 1 x 10(6) cells from 34 to 40 passages seeded onto nine BAM scaffolds were cultured for one week. Nine other scaffolds were left unseeded. Scaffolds were grafted into a surgically created defect within the anterior bladder wall: nine rats with acellular matrices and nine with cell-seeded BAM. Rats observed for six months were killed in monthly intervals. We performed gross examination, X-ray cystography, and hematoxylin eosin, cytokine (CK)-7, CK-20, myoglobin, and desmin staining of the excised bladders. RESULTS: Minimal adhesions were observed and urinary leakage was noted in one case. Two animals died in the acellular group. Rats developed stone disease in bladders reconstructed with acellular BAM. Bladder capacity was similar, but the shape was regular and characteristically oval only in bladders grafted with cell-seeded BAM. Muscle layers in the apical parts of the reconstructed bladder walls were extremely thin in the cases of acellular grafts and thicker in bladders reconstructed with cell-seeded grafts. Muscle layer regeneration was better in the cell-seeded group. Urothelium regenerated in all animals. CONCLUSIONS: We have shown that hair follicle stem cells may be used for rat bladder wall regeneration. PMID- 20005398 TI - Multipotent progenitor cells derived from human umbilical cord blood can differentiate into hepatocyte-like cells in a liver injury rat model. AB - Umbilical cord blood (UCB), a rich source of hematopoietic stem cells, offers practical and ethical advantages. It has been reported that various adult stem cells transplanted into a damaged liver show characteristics of a hepatic lineage. In a previous study, we reported on novel UCB-derived adult stem cells, termed umbilical cord blood-derived multipotent progenitor cells (UCB-MPCs). We demonstrated that these cells were capable of differentiating into hepatocyte- like cells in vitro. To assess the hepatic differentiation capacity of UCB-MPCs, rat models of hepatic injury were generated using carbon tetra-chloride (CCl(4)) with transplantation of cells into the liver. The transplanted cells successfully incorporated into the liver of the recipient animal differentiated into functional hepatocyte-like cells that expressed hepatocyte-specific markers, such as CK-18 and albumin. Moreover, human albumin was detected in the serum of the recipient rat model. These data indicated that UCB-MPCs were capable of displaying similar characteristics to those of functional hepatocytes in a recipient liver. UCB-MPCs may prove to be a useful, transplantable alternative for hepatic progenitor cells in both experimental and therapeutic applications. PMID- 20005399 TI - Endothelial nitric oxide synthase (eNOS) gene polymorphism in early term chronic allograft nephropathy. AB - Chronic allograft nephropathy (CAN) is a complex phenomenon caused by underlying kidney disease with superimposed enviromental and genetic factors. CAN development begins with progressive renal microvascular injury. Endothelial cells play key roles in the regulation of vascular tone, permeability, and remodeling. A reduction in basal nitric oxide (NO) release as a result of genetic variation in endothelial NO synthase (eNOS) function may predispose to hypertension, thrombosis, vasospasm, and atherosclerosis, all contributing to the development of CAN. We analyzed the G894T mutation at exon 7 of the eNOS gene in relationship to CAN among 81 children with renal transplantations. The 20 patients who developed CAN underwent renal biopsies for histological confirmation. Proteinuria and hypertension were observed in CAN. We selected 173 healthy reference subjects. The G894T polymorphism of the eNOS gene was determined by PCR restriction fragment-length polymorphism analysis. The group included 33 male and 48 female subjects who received 32 living-related grafts and 49 from deceased donors (DD) donors. Donor age (y) was 32.7 +/- 13.7 and the HLA A,B,DR mismatch number of the cadaveric cases was 3.5 +/- 0.79. The distribution of the genotypes were ENOS GG/GT/TT 48%, 33%, 19%, respectively. G-alleles frequency was 64.8%; T allele frequency was 35.2%. ENOS G894T gene polymorphism did not seem to influence long-term renal allograft outcome. Recipient ENOS G894T gene polymorphism did not alter the risk of chronic allograft failure. Even if NO synthesis and bioactivity are influenced by this polymorphism, many vasoactive factors may have roles to suppress the advantageous effects of NO. PMID- 20005400 TI - Detection of citrate synthase autoantibodies in rats with chronic allograft nephropathy. AB - Citrate synthase (CS) is the one of the key enzymes in the citric acid cycle and an important mitochondrial autoantigen. The autoimmune responses against CS have not been studied in chronic allograft nephropathy (CAN). This study investigated the role of specific CS autoantibodies in rats bearing renal allografts affected with CAN. METHODS: Fisher344 rat renal grafts were orthotopically transplanted into Lewis rats following the procedure of Kamada with our modification. Lewis-to Lewis and Fisher344-to-Fisher344 kidney transplantations were also performed as autologous control groups (each n = 9). All the allograft recipients given cyclosporine (10 mg/kg(-1)d(-1) x 10 d) were divided into four groups (each n = 9): (1) vehicle: normal saline orally; (2) cyclosporine: 6 mg/kg(-1)d(-1); (3)FK506: 0.15 mg/kg(-1)d(-1); (4) mycophenolate mofetil (MMF): 20 mg/ kg(-1)d( 1). At 4, 8, and 12 weeks posttransplantation, the animals were sacrificed to harvest sera and renal allografts. The serum creatinine (SCr) was measured and pathological changes assessed according to Banff 97 criteria. IgM and IgG isotypes of CS antibodies were detected in all recipient sera by enzyme linked immunosorbent assays. RESULTS: Both IgM and IgG isotype CS autoantibodies were observed in the sera of all the recipients before and after transplantation, but the levels of IgM CS autoantibody were obviously higher than IgG isotype in all the blood samples. It was stable not only in autologous but also in allograft groups. In both autologous groups, the SCr and IgM and IgG isotype CS autoantibodies showed no obvious change before and after transplantation, and no typical CAN occurred. The values of IgG isotype of CS autoantibody (DeltaOD) at 4, 8 and 12 weeks were stable. At 4 weeks, the values of SCr, Banff score, and IgG isotype CS autoantibody (DeltaOD) were not significantly different (P > .05) among the allograft groups. At 8 and 12 weeks, with progression of CAN in vehicle, cyclosporine and FK506 groups' values of SCr, Banff score, and IgG (DeltaOD) also increased dramatically (P = .005) in all three groups when compared with the baseline and 4 week values, but the differences among the three groups were not significant (P > .05). At 8 and 12 weeks, the MMF group suffered mild-to-moderate CAN, but the values of SCr and Banff score were significantly lower than those in the other three groups. MMF significantly inhibited the formation of IgG (DeltaOD) when compared with the other three groups (P = .02). CONCLUSION: This study suggested that the IgG isotype of CS autoantibody contributes to CAN after kidney transplantation. The IgM isotype is physiological. MMF significantly inhibited the formation of IgG isotype CS autoantibody, which may be related to its effects to alleviate CAN. PMID- 20005401 TI - Gene profiling of cyclosporin-enhanced transitional cell carcinoma in rat model. AB - OBJECTIVE: We sought to study the effect of cyclosporine (CsA) on development of malignancy. MATERIALS AND METHODS: The observation was performed in a rat model, in which transitional cell carcinoma of urinary bladder was induced with N-butyl N-(-4-hydroxybutyl) nitrosamine. CsA was added in the food for the rats. At the end of 30 weeks, we examined the tumor burden in the urinary bladders, and compared gene expressions between the CsA-enhanced and non-CsA-enhanced tumor groups by gene profiling. RESULTS: CsA feeding increased tumor burden: 2.3 +/- 0.9 versus 1.1 +/- 0.5 g (P < .05). Gene profiling showed many variations involved in CsA enhanced malignant development. Twenty-three genes with known functions were upregulated, and 46 genes with known functions downregulated. In all, 111 genes were involved in the CsA-enhanced malignant development. The regulated genes in the present study constituted 23 pathways mostly involved in carcinogenesis. CONCLUSIONS: CsA plays an important role in tumor development through gene regulation, which may constitute pathways to malignant progression. PMID- 20005402 TI - Tranilast attenuates chronic cyclosporine nephrotoxicity in rats. AB - Our aim was to explore the effects of the anti-allergic and antifibrotic agent tranilast on chronic cyclosporine (CsA) nephrotoxicity in rats. METHODS: Eighteen Sprague-Dawley rats were randomized to be given the following daily treatments by gavage for 4 weeks: (1) controls, olive oil; (2) CsA group, CsA 25 mg/kg; (3) CsA plus tranilast group, CsA 25 mg/kg and tranilast 400 mg/kg. We examined the body weights and the effects of tranilast on histopathology, macrophage (Mvarphi) infiltration, and expression of osteopontin (OPN). RESULTS: The administration of tranilast improved body weight gain by CsA-treated rats (232 +/- 24 vs 203 +/- 6 g; P < .05). This treatment reduced the expression of OPN protein and infiltration of macrophages (9.14 +/- 2.7 vs 22.44 +/- 5.68 ED-1 positive cells per high power field, P < .05). Furthermore compared with the CsA alone group, it ameliorated tubulointerstitial fibrosis scores (1.18 +/- 0.08 vs 2.57 +/- 0.21, P < .05). CONCLUSION: Tranilast attenuated tubulointerstitial fibrosis through decreased expression of OPN protein and macrophage infiltration, showing renal protective effects in rats with chronic CsA nephrotoxicity. PMID- 20005403 TI - Hemodynamic monitoring in pigs undergoing orthotopic abdominal multivisceral transplantation. AB - Anesthesia for abdominal multivisceral transplantation (MVTx) is challenging. Surgical success depends on hemodynamic stability throughout organ removal from the donor and engraftment in recipients. The aim of the current study was to summarize our experience with anesthetic management including monitoring of hemodynamic changes in pigs during MVTx procedures. MATERIALS AND METHODS: Ten male pigs were randomly divided into two groups: donors and recipients for five MVTxs. A pulmonary artery catheter (PAC), transesophageal echocardiography (TEE), and pulse indicator continuous cardiac output (PiCCO) were used intraoperatively in recipients to monitor hemodynamic changes. RESULTS: Two recipients had cardiac arrest and died intraoperatively presumably due to hypovolemia. The data obtained by PAC and TEE from three successful cases showed huge changes in hemodynamics during the stages of organ removal and reperfusion after engraftment. Use of PiCOO in one of those three cases allowed successful fluid management, resulting in stable intraoperative hemodynamics. CONCLUSION: The anesthetic management for MVTx surgery was improved by PAC, TEE, and PiCOO monitoring. PMID- 20005404 TI - A normothermic perfusion bioreactor to preserve viability of rat groin flaps extracorporally. AB - INTRODUCTION: Various attempts have been made to prolong tissue survival ex vivo. To achieve an adequate ex vivo condition for flap perfusion at normothermic temperatures in a bioreactor model, a suitable perfusion solution is necessary. The main purpose of our setting at 37 degrees C was to produce conditions under which multilineage stem cells from adipose tissue could differentiate. This study was designed to evaluate the effect of permanent perfusion on fat flaps of the rat. MATERIALS AND METHODS: We elevated an epigastric adipofascial flap based on the inferior superficial epigastric vessels bilaterally in male Lewis rats and connected it to a bioreactor. The system was run by a cable pump and filled either with Hannover or Eurocollin's solution with or without permanent perfusion for 10 days. The lactate dehydrogenase (LDH) level in each solution was analyzed every 48 hours, assuming that injured cells emit this enzyme to the extracellular space and consequently to the perfusion solution. Histological samples were analyzed at the end of each trial. RESULTS: There was a continuous significantly greater LDH level (P < .001) in bioreactors perfused with Hannover than with Eurocollin's solution. The nonperfused bioreactors showed a similar finding with lower levels compared with their perfused equivalents. Histological examination revealed significantly better preserved (P < .001) fat tissue structures in Hannover solution-perfused specimens. CONCLUSION: Because LDH has a half life of 24 hours, ongoing production of this enzyme for 10 days is a marker for an injured tissue consisting of viable cells. Bioreactors run with Hannover solution showed significantly higher LDH levels. Histological analyses revealed intact cells preserved in Hannover solution. Thus, Hannover solution seemed to be superior to Eurocollin's solution to keep flaps viable under normothermic conditions. The presented model facilitated fat tissue conservation under normothermic conditions and represented a foundation for further studies on the differentiation of vascularized fat tissue. PMID- 20005405 TI - Investigation of porcine endogenous retrovirus in the conservation population of Ningxiang pig. AB - Porcine endogenous retrovirus (PERV) varies between pig breeds. Screening and analysis of PERV in putative pig breeds may provide basic parameters to evaluate the biological safety of xenotransplantation from pigs to humans. In this study, PERV was investigated among the conservation population of the Ningxiang pig. The result revealed that the genotype of PERV distribution was subtype A, 100%; subtype B, 100%; and subtype C, 100%. The env sequences of PERV-A and -B showed 11 clones detected by KpnI and MboI digestion, indicating that there existed multiple variants of PERV-A and -B in the Ningxiang pig. Reverse transcriptase polymerase chain reaction results showed that PERV had transcriptional activity in these individuals. In addition, PERV A/C recombinant was detected in most individuals of Ningxiang pig. Because PERV A/C recombinants increase the potential infectious risk, the breed may not be a proper donor for xenotransplantation. PMID- 20005406 TI - Effects of radiation on protein oxidation and lipid peroxidation in the brain tissue. AB - Radiation produces reactive oxygen species that modify macromolecules such as protein and lipid by oxidation and act in many pathological processes, causing serious damage to the brain. This study sought to evaluate the effects of radiation and the protective effect of N-acetylcysteine (NAC) on the brain tissue of guinea pigs based on the levels of lipid peroxidation and protein oxidation. Thirty-two guinea pigs were divided into groups of eight animals each: control group (group I); radiation group (group II); NAC group (group III), and NAC administered before radiation exposure group; (group IV). Guinea pigs in groups II and IV were exposed to Co(60) radiotherapy using the Source-Axis-Distance = 80 technique. The levels of protein carbonyl content and malondialdehyde (MDA), which is a marker for lipid peroxidation, were investigated by the Evans-Levine and Uchiyama-Mihara methods, respectively. The protein carbonyl and MDA levels of group II were significantly greater than those of group I (P < .01). The protein carbonyl and MDA levels of group IV were lower than those of group II. The difference between the MDA levels of group IV and group II was significant (P < .01); however, the difference in protein carbonyl levels between the two groups was not significant. The results indicated that radiation increased protein oxidation and lipid peroxidation in the brain, and NAC administration before radiation exposure may aid in the treatment by decreasing protein and lipid oxidation. PMID- 20005407 TI - Application of a self-made swivel intravenous transfusion device in constructing allogenic small bowel transplantation rejection model in rats. AB - BACKGROUND: Lacking a long-term continuous intravenous infusion device for experimental animals is the biggest obstacle to constructing an allogenic small bowel transplantation rejection model. OBJECTIVE: To establish a rat heterotopic small bowel transplantation model with the use of a swivel device for intravenous transfusion. METHODS: The randomized block design of controlled observations included 90 inbred male F344/NCrl BR rats as donors and 90 LEW/Crl rats as recipients. On the basis of body weight, the rats were divided into three groups each with 30 pairs of rats. Group 1, the control group, underwent small bowel transplantation. Group 2, the transfusion group, received small bowel transplantation and continuous infusion of parenteral nutrient solution. Group 3, the tacrolimus (FK506) treatment group, received small bowel transplantation, continuous infusion of parenteral nutrient solution, and intravenous injection of FK506. The general status and survival time of transplanted rats were observed for 5 weeks. RESULTS: The operative success rate was 100%. The survival rate was 83.3% (25/30) in the control group; and 96.7% (58/60) in groups 2 and 3 (P = .039). The mean survival time of the FK506 group was 22.1 +/- 13.4 days, which was significantly higher than that in the transfusion group (10.4 +/- 2.9 days, P = .023). CONCLUSION: The simple, practical swivel intravenous infusion device provided treatment for more than 30 days, a useful tool for heterotopic segmental small bowel transplantation. PMID- 20005409 TI - Bortezomib successfully reverses early recurrence of light-chain deposition disease in a renal allograft: a case report. AB - Light-Chain Deposition Disease (LCDD) frequently recurs after renal transplantation, displaying a pernicious course. Herein we have described a 39 year-old Caucasian man with a history of immunoglobulin G-kappa multiple myeloma who failed two chemotherapy regimens, but ultimately responded to the combination of thalidomide, bortezomib, and dexamethasone followed by high-dose melphalan and autologous stem cell transplantation 3 years prior to transplantation, during which time he showed no evidence of persistent or recurrent disease. At 3 days following spousal living related renal transplantation, he displayed a rapid deterioration of renal function requiring dialysis therapy. This episode failed to respond to empiric antirejection therapy including anti-thymocyte globulin, plasmapheresis, and anti-CD20 monoclonal antibody. Increasing evidence suggested recurrence of LCDD, including positive immunofluorescence staining of basement membranes and vessels for kappa light chains as well as free kappa light chains in his urine and serum. Following suspension of sirolimus, he was initiated on and responded to bortezomib (1.3 mg/m(2)) with discontinuation of dialysis within 3 weeks and progressively improving renal function. His maintenance therapy, in addition to six 2-week-long cycles of bortezomib separated by 1-week rest periods, includes cyclosporine (50 mg twice daily), prednisone (10 mg daily), and curcumin (9 g daily). In sum, bortezomib rescue therapy salvaged a spousal renal transplant afflicted with recurrent LCDD. PMID- 20005408 TI - Expression of mesenchymal, hematopoietic, and biliary cell markers in adult rat hepatocytes after partial hepatectomy. AB - BACKGROUND AND PURPOSE: It has been suggested that liver regeneration can occur either by differentiated adult hepatocytes which retain the capability for several rounds of replication or by hepatic progenitor cells, depending on the number of hepatocytes lost. We sought to study the differentiation potential of hepatocytes following partial hepatectomy (PH) in rats. METHODS: Using immunohistochemistry and confocal microscopy we studied the distribution of cytokeratin 7 (CK7), CK19, vimentin, desmin, CD34, and c-kit among adult rat liver hepatocytes after PH at various times just after hepatectomy and after 8, 16, 24, 36, 48, and 60 hour and 6 and 16 days. RESULTS: Vimentin, c-kit, and desmin positivity were observed in regenerating hepatocytes in the early stages. Desmin and vimentin staining were also demonstrated in stellate cells. Staining enhancement in stellate cells progressed from day 3 to day 6. No liver sections were stained positive for CD34, CK19, or CK7. CONCLUSION: After PH, mature hepatocytes revealed their potential to regain the markers that they do not express when they are quiescent. This result supported the plasticity and differentiation potential of adult hepatocytes during regeneration. PMID- 20005410 TI - Postexposure prophylaxis of H1N1 with oseltamivir in a newly transplanted kidney recipient receiving intense immunosuppressive therapy. AB - Solid organ transplant recipients undergoing immunosuppressive therapy are considered to be at high risk for serious infectious complications. Recently in the United States, a pandemic of H1N1 flu infection has been reported with serious complications. We describe H1N1 infection in a living kidney donor and the 42-year-old kidney transplant recipient exposed to this kidney donor and undergoing intense immunosuppressive therapy. Postexposure prophylaxis with oseltamivir was effective to prevent H1N1 influenza A virus in a donor and a recipient. PMID- 20005411 TI - Myocardial metastasis of cutaneous squamous cell carcinoma in a renal transplant recipient. AB - Myocardial metastasis from a cutaneous squamous cell carcinoma (SCC) is rare. Herein we have presented a case of metastasis from cutaneous SCC to the myocardium in a renal transplant recipient, which was confirmed by a cardiac fine needle biopsy. Postmortem examination revealed disseminated metastatic disease involving myocardium, lungs, thyroid, skin, and peritoneum secondary to cutaneous SCC likely related to immunosuppression. At 46 years of age, he received a renal transplant for chronic renal failure caused by chronic glomerulonephritis. He started to develop multiple nonmelanoma skin cancers 4 years later. At least 23 invasive SCCs and 14 basal cell carcinomas were excised. His immunosuppressive regimen consisted of cyclosporine (150 mg), azathioprine (75 mg), and prednisone (10 mg daily), which was not modified despite multiple nonmelanoma skin cancers. Our case report further illustrates the potentially aggressive and fatal nature of cutaneous SCCs that can develop in organ transplant recipients. It argues for modification of the immunosuppressive regimen in such patients. The management of renal transplant patients with nonmelanoma skin cancers remains difficult and complex. PMID- 20005412 TI - Hemorrhagic cystitis secondary to adenovirus or herpes simplex virus infection following renal transplantation: four case reports. AB - Viral infections are common complications following renal transplantation. However, there have been few reported cases of viral cystitis secondary to herpes simplex virus or adenovirus infection. Herein, we have reported four cases of hemorrhagic cystitis secondary to infections with herpes simplex virus and adenovirus following renal transplantation. The etiology was adenovirus in three cases and herpes simplex virus in the remaining case. In all four cases, the primary cause of the renal dysfunction was diabetic nephropathy. All four patients presented with a clinical profile characterized by dysuria, pollakiuria, macroscopic hematuria, and graft dysfunction. Three of the four patients developed these symptoms within the first 3 months after renal transplantation. In all four cases, there was an increase, albeit slight, in creatinine levels, which returned to normal or near-normal values upon resolution of the symptoms. Acute cellular rejection was observed in only one case. Although rare, hemorrhagic cystitis secondary to infection, which typically occurs early in the posttransplant period, causes pronounced symptoms. The infection appears to be self-limiting, resolving completely within 4 weeks. PMID- 20005413 TI - Successful kidney transplantation from a donation after cardiac death donor with an ileal conduit. AB - Kidney transplantation is the treatment of choice for those affected by end-stage renal disease. Consent for organ donation continues to be one of the greatest challenges to transplanting more patients waiting for a life-saving transplant. In an attempt to increase the donor pool for patients on kidney transplant waiting lists, transplant surgeons and physicians have expanded their acceptance criteria to include expanded criteria donors, donation after cardiac death donors, as well as those donors who present unique technical challenges to organ recovery. Here we report a successful kidney transplant from a kidney donor who died from cardiac causes and who previously underwent an ileal conduit for a neurogenic urinary bladder secondary to a spinal cord injury. PMID- 20005414 TI - Sarcomatoid renal cell carcinoma in a renal transplant recipient. AB - The incidence of transplanted kidneys derived from elderly donors is increasing because of the larger waiting lists and greater age of patients with end-stage renal failure. Compared with young donors, one of the problems is the heightened risk of neoplasm transmission. We report 2 cases of kidney recipients, both of whom developed a sarcomatoid renal cell carcinoma after receiving a kidney transplant from the same 68-year-old male donor, who did not show signs of a neoplasm on a previous abdominal ultrasound or a pretransplant biopsy. The first recipient was a 66-year-old woman who developed a kidney mass with several urologic obstructive complications, tumor dissemination, and death at 9 months after kidney transplantation. The second recipient was a 48-year-old asymptomatic man with normal renal function, who was studied after the results of the first recipient, revealing another renal tumor. Transplant nephrectomy was performed and a peritoneal implant was resected. The patient is alive without evidence of a neoplasm after 18 months. Herein we have discussed the mechanisms of neoplasm transmission in kidney transplantation and possible strategies for its prevention and treatment. PMID- 20005415 TI - False serologic evidence for acute primary toxoplasmosis during liver transplantation for fulminant hepatitis B: a case report. AB - Acute primary Toxoplasma gondii infection is usually considered to be a contraindication for solid organ transplantation. Recent reports of acute T. gondii infection have highlighted the need to include T. gondii serology in the pretransplant screening of solid-organ transplant recipients. However such serology might be misleading. We describe the case of a 25-year-old woman who received a liver transplantation for life-threatening liver failure due to hepatitis B virus infection. The presence of high IgM titers against T. gondii, as detected by membrane immunoassay, immunofluorescence, and mu-capture ELISA tests, together with the absence of IgG antibodies in the immediate pretransplant serology screening suggested acute primary T. gondii infection at the time of transplantation. We initiated a preemptive therapy with intravenous clindamycin and cotrimoxazole. However, negative PCR and IgA capture assays, together with the absence of a sustained IgG response finally excluded the initial diagnosis of primary toxoplasmosis, leading to discontinuation of antitoxoplasmosis therapy. This case illustrates the problem that, in the context of fulminant hepatitis B, serologic markers for acute primary toxoplasmosis can be falsely positive. Confirmation by PCR and IgA antibody determinations is required to confirm this diagnosis. PMID- 20005416 TI - Emergency liver transplantation after umbilical hernia repair: a case report. AB - We report a rare case of acute liver failure due to embolization of the liver after an umbilical hernia repair in a patient with Child B liver cirrhosis and status posttransjugular intrahepatic portosystemic shunt (TIPSS). This patient initially presented with a symptomatic umbilical hernia. His umbilical vein was open (Cruveilhier-Baumgarten syndrome). After hernia repair the patient developed thrombosis of the umbilical vein with consequent partial embolization to, and acute failure of, the liver. The patient underwent successful emergency liver transplantation. This disease needs close collaboration among surgeons, gastroenterologists, hepatologists, radiologists, nutritionists, and transplant teams to establish an effective treatment plan. PMID- 20005417 TI - Graft-versus-host disease presenting with pancytopenia after en bloc multiorgan transplantation: case report and literature review. AB - Graft-versus-host disease (GVHD) is a dreaded complication of bone marrow and solid organ transplantation. Commonly affected organs include skin, liver, and the gastrointestinal tract, with bone marrow and renal involvement occurring more rarely. GVHD is less commonly seen with solid organ transplants. Fewer than 100 cases of GVHD have been reported in the literature following liver transplantation. We report a case of a 53-year-old woman who required a multiorgan transplant after a complicated postoperative course following paraduodenal hernia repair. She developed isolated pancytopenia approximately 4 months after receiving an en bloc transplant involving the liver, kidney, small bowel, and pancreas. No evidence of skin, gastrointestinal, or hepatic involvement was discovered. HLA typing of the peripheral blood revealed that 28% of patient peripheral blood was composed of donor lymphocytes. Bone marrow biopsy showed a markedly hypocellular marrow with 23% donor lymphocytes and 80% of the T cell population from the donor as well. The patient began treatment for GVHD, including high-dose steroids, basiliximab, and rituximab. Unfortunately, she developed overwhelming sepsis and subsequently died. This case describes an instance of GVHD manifested by isolated pancytopenia after en bloc transplantation of multiple solid organs. GVHD is a rare, but serious complication of solid organ transplantation that can result in death. Although isolated bone marrow involvement is uncommon, it must be considered early to avoid a delay in diagnosis. This case also highlights an association of GVHD with multiorgan transplants, although this is incompletely characterized in the current literature. PMID- 20005418 TI - Geriatrics in the most aged country, Japan. AB - The aging of society is a common problem in many developed countries. To tackle the problems related to an aging society, the role of geriatricians, as well as government support, is becoming more and more important. There is need to recruit young physicians with the skills required to care for elderly patients, and to establish an education system which encourages more young physicians to undertake training in geriatrics. Additionally, there is a need for improvement of our insurance systems to support such care. Our approach to the aging society will pave the way for other countries in Asia and the west. PMID- 20005419 TI - Pain and health-care utilization among older men in a veterans care home. AB - Pain is a common health-care issue, and the prevalence increases with advancing age. Although it is often assumed that people with chronic pain are associated with a higher consumption of health care, evidence supporting this assertion is insufficient. Data from the Longitudinal Older VEterans (LOVE) study were stratified to explore the prevalence of pain and its relationship with health care utilization. In total, data from 574 residents (mean age: 80.9+/-5.4 years, all male) were obtained. Among them, 92.8% were physically independent and 20.2% of them had mild to moderate cognitive impairment. Overall, 153 (26.3%) subjects reported pain; 114 (74.5%) subjects with mild pain and the remaining 39 (25.5%) subjects with moderate pain. The most commonly reported pain was lower back pain (40.5%, 62/153), which was followed by joint pain (29.4%, 45/153). Subjects with pain were more likely to have higher scores on the Geriatric Depression Scale (2.4+/-2.4 vs. 1.8+/-2.2, p = 0.023) and care-complexity problems (4.7+/-2.0 vs. 3.9+/-1.9, p < 0.001), despite being similar in age (81.3+/-5.0 vs. 80.8+/-5.5, p = 0.271), cognitive status and physical independence. Compared with pain-free subjects, subjects with pain were more likely to be hospitalized in the 12-month study period (0.71+/-1.20 vs. 0.46+/-1.00, p = 0.010), but the utilization of emergency department treatment (1.74+/-1.23 vs. 1.88+/-1.63, p = 0.560) was not statistically significant. In conclusion, the prevalence of pain among residents in a Taiwanese veterans care home was 26.3%; subjects with pain having more depressive symptoms, higher clinical-care complexity, and more likely to be hospitalized during the 12-month follow-up. PMID- 20005420 TI - Geriatric education for the physicians of tomorrow. AB - The world's population is aging and there is need for more geriatricians. Current training programs, however, are not producing a sufficient number of geriatricians to meet that need, largely because students and residents lack interest in a career in geriatrics. A variety of reasons have been suggested to explain that lack of interest, and several changes in geriatrics training might increase the number of medical trainees who choose a career in geriatrics. These changes include recruiting medical students who are predisposed to geriatrics, loan forgiveness programs for those who enter careers in geriatrics, increased reimbursement for geriatric care, providing geriatric education to physicians in all specialties throughout their training, and refocusing geriatrics training so it includes the care of healthy vigorous older adults, rather than an exclusive focus on those with debility and chronic or fatal illnesses. PMID- 20005421 TI - The development of intermediate care services in England. AB - The origins of intermediate care as a health policy in England are reviewed. The randomized controlled trial (RCT) evidence for the various intermediate care service models is discussed from the perspectives of clinical, service and economic outcomes. This evidence base suggests that the hospital-at-home approach is currently the best RCT supported intermediate care service model. It is argued that intermediate care in England has yet to fulfil its expectations. Changing staff skills and attitudes has proved a challenge, and the integration of intermediate care with mainstream services has been especially difficult. New national guidance has been produced that is attempting to refocus intermediate care its intended target group of frail older people, and not to specifically exclude older people with mental health problems. PMID- 20005422 TI - First-ever ischemic stroke in Taiwanese elderly patients: predicting functional independence after a 6-month follow-up. AB - The incidence of ischemic stroke increases with age, and it has a great impact on patients' functional independence. The aim of this study was to analyze the clinical features, laboratory findings, and stroke subtypes in different age subgroups and identify the predictive factors for functional independence 6 months after stroke. A total of 533 patients with first-ever ischemic stroke were enrolled in this study. They were divided into two subgroups: more than 80 years old (n = 108) and less than 80 years old (n = 425). Patients aged 80 years or over had higher frequencies of heart disease and atrial fibrillation, and lower frequencies of dyslipidemia, alcohol drinking, and a family history of ischemic stroke. Significantly lower body mass index, serum albumin levels, and lipid profiles, including total cholesterol, low-density lipoprotein, and triglyceride levels, but higher severity of initial neurologic deficit, and higher rates of mortality and complications during hospitalization were noted in patients aged over 80 years. The multivariate logistic regression analysis showed that higher serum total cholesterol level, less severity of neurologic deficit at admission, and absence of a history of diabetes mellitus were predictive of functional independence 6 months after stroke. PMID- 20005423 TI - Towards age-friendly hospitals and health services. AB - In this paper, we reviewed rationale for an age-friendly hospital (or health service) and propose our framework in detail. In a rapidly aging society, development of policies and programs to optimize people's wellbeing and function as they age is an urgent issue. Most older persons extensively use health care. Unfortunately, current practices are unfriendly and may even add risks to the older person. Health-promotion interventions delivered in clinical settings and management of admissions have been shown to be associated with better outcomes. We developed Taiwan's Framework of Age-Friendly Hospitals to address the responsibility of health-care organizations and provide systematic guidance on hospital management policy, communication and services, physical environments, and care processes. This framework takes a life-course perspective and population approach and includes evaluation and quality improvement as an integral part of an age-friendly initiative. The diffusion and adaptation of age-friendly practices in hospitals can be monitored and its impact evaluated in the future. PMID- 20005424 TI - Emergency department (ED) utilization and outcome of oldest old men presenting with geriatric syndromes in a veterans care home in Taiwan. AB - The first presentation of elderly people in the emergency department (ED) is commonly nonspecific and atypical, often in the form of geriatric syndromes, i.e. falls, immobility, incontinence, or deteriorating mental function. The purpose of this study was to evaluate the management and outcomes of institutionalized elderly people who initially presented with geriatric syndrome (GS) in the ED. A retrospective chart review of Banciao Veterans Care Home residents who visited the ED of a tertiary medical center was done. Demographic data including age, sex, modes of arrival, category of triage, time of visit, main presenting symptoms, principal diagnosis, medical expenditures, and clinical outcomes were recorded. From January to December, 2006, 629 ED visits (mean age, 82.1+/-5.3 years, all male) were retrieved. The overall prevalence of GS was 23.8%. When GS subjects were admitted, they were more likely to be transferred to step-down community hospitals for post-acute care (OR = 2.63; 95% CI: 1.36-5.08, p = 0.004). GS was common in institutionalized elderly people calling for ED services, and GS subjects were more likely to be transferred to step-down community hospitals after hospitalization. Comprehensive geriatric assessments may be of value for institutionalized elderly patients visiting the ED. PMID- 20005425 TI - Association between metabolic syndrome (MS) and bone mineral loss: a cross sectional study in Puli Township in Taiwan. AB - Results of studies regarding the association between the MS and osteoporosis have been largely controversial. The objectives of this study were to measure bone mineral density (BMD) with and without the MS and evaluate the associations between bone mineral loss and the components of the MS. In all, 352 men (mean age 70.6+/-11.9 years) and 468 women (mean age 67.8+/-12.0 years) in Puli township, Taiwan, were enrolled. Height (cm), weight (kg), waist circumference (cm), and blood pressure (mmHg) were measured. Fasting plasma glucose (FPG) and blood lipids were determined. Calcaneal BMD was measured using the ultrasound pulse transmission method. The prevalence of MS was 33.0% in men and 44.2% in women. In subjects with the MS, lower diastolic blood pressure (DBP) in both sexes (p < 0.001), lower triglycerides (p = 0.016) and more central obesity in men (p = 0.011) predicted bone mineral loss. The MS was not associated with bone mineral loss in either of sexes (p = 0.550 in men; p = 0.628 in women). PMID- 20005426 TI - Plasminogen activator inhibitor type 1 (PAI-1) is a valuable biomarker for predicting the metabolic syndrome (MS) in institutionalized elderly residents in Taiwan. AB - Circulating levels of inflammatory and prothrombotic factors are elevated in the metabolic syndrome (MS) and linked with the occurrence of cardiovascular events. The aim of our study was to investigate the relationship between inflammatory and prothrombotic markers and the MS in elderly institutionalized residents. A total of 326 non-diabetic residents of Chuang-Hua Veterans Care Home (age: 79.9+/-4.1 years; 100% males) were enrolled. MS was diagnosed according to the AHA/NHLBI Scientific Statement criteria. Body fat percentage was measured by bioelectrical impedance analysis. Insulin resistance was calculated by homeostasis model assessment for insulin resistance (HOMA-IR). Inflammatory markers, including tumor necrosis factor-a (TNF-alpha), high sensitivity C-reactive protein (hsCRP), and plasminogen activator inhibitor-1 (PAI-1), were determined using ELISA. Elderly residents with the MS had higher systolic and diastolic blood pressures (both p < 0.001) and higher HOMA-IR (p < 0.001), hsCRP (p = 0.008), and PAI-1 levels (p < 0.001) than those without the MS. On multivariate logistic regression analysis, PAI-1 was an independent risk factor for the MS. Of the MS components, elderly residents with higher waist circumferences and higher levels of plasma fasting glucose, and triglyceride (TG), and lower levels of high density lipoprotein (HDL) had higher PAI-1 levels than those without the above components. PMID- 20005427 TI - Relationship between hyperuricemia (HUC) and metabolic syndrome (MS) in institutionalized elderly men. AB - This study investigated the relationship between hyperuricemia (HUC) and the components of the metabolic syndrome (MS) among elderly institutionalized men. In addition, this study explored the relationship between HUC and serum inflammatory markers. A total of 333 participants from Chang-Hua Veterans Care Home were enrolled. The MS was defined using a modified ATP III definition issued in 2004 by the Bureau of Health Promotion, Department of Health, ROC (Taiwan). The participants' mean age was 78.6+/-3.9 years, and their mean serum uric acid level was 6.9+/-1.7 mg/dl. The prevalence of HUC was 46.2% (n = 154). The prevalence of the MS was 38.4% (n = 128). HUC was correlated with components of the MS, including waist circumference (WC), triglyceride (TG), and high density lipoprotein cholesterol (HDL-C) but it was not related to blood pressure (BP) and fasting plasma glucose (FPG). Moreover, increased serum creatinine, albumin, prealbumin, and body fat were also associated with HUC. The plasma activator inhibitor-1 (PAI-1) levels were significantly elevated in the HUC group, but serum interleukin-6 (IL-6), tumor necrosis factor-alpha (TNF-alpha), serum intercellular cell adhesion molecule-1 (sICAM-1), serum levels of vascular cell adhesion molecule-1 (sVCAM-1), and P-selectin were not related to HUC. HUC in elderly men may represent poorer renal function, better nutritional status, and increased body fat. PMID- 20005428 TI - Clinical features of benign paroxysmal positional vertigo (BPPV) in Taiwan: differences between young and senior age groups. AB - BPPV is a common cause of vertigo. Several treatment procedures can facilitate recovery. In this study, we aimed to identify the demographic features, resolution and recurrence rates and impacts on daily activities in BPPV patient between young and senior age groups in Taiwan. This retrospective study recruited 218 patients of BPPV. Medical history, canal involvement, treatment required for complete resolution, symptom free period and recurrence rates were evaluated between the two age groups. Up to 80.7% of patients were successfully treated by a single treatment. For patients aged more than 65 years, the recurrence rate was 1.7 times higher than that in the younger age group (p = 0.07). The symptom-free period before recurrence was nearly 2.2 times longer in the senior age group (p = 0.03). Work-related activities were influenced more by BPPV in the younger age group (p = 0.03). We conclude that BPPV is prone to occur and recur in people of senior age. Clinicians should have the knowledge to diagnose different types of BPPV and treat it accordingly to prevent further complications. PMID- 20005429 TI - Development of services for older patients with falls and fractures in England: successes, failures, lessons and controversies. AB - Falls and fragility fractures are becoming a major epidemic of aging, with each year around one third of people aged 65 and over, and half of people over 80 years, experiencing at least one fall. This has major implications, both for older people themselves, and for health- and social-care services. There is evidence for benefits to be gained from interventions to manage fracture patients better, responding to and preventing falls, identifying and treating those with bone fragility, and to maximizing population health by preventing or delaying frailty. In addition, the most consistently proven way of ensuring that patients with fracture receive evidence-based secondary prevention in practice is through the use of specialist services linking hospital-based fracture services with follow-up on patient discharge. Here we describe the evolution of the approach taken by the English National Health Service (NHS) over the past decade to address this issue, along with the successes, failures and lessons potentially relevant to other health systems. Approaches used have included national guidelines, national audits and involvement of Department of Health Resources to drive up quality. The key themes are "responding to the first fall to prevent the second", "better recognition and treatment for osteoporosis", "responding to first fragility fracture to prevent the second" and "better interdisciplinary management of patients admitted with hip fracture", and the audit systems which now exist to describe changes in practice and the outcomes which follow. PMID- 20005430 TI - Developing geriatric services and research in Taiwan. Preface. PMID- 20005432 TI - [Glyoxal: a possible polyvalent substitute for formaldehyde in pathology?]. AB - The quest for formaldehyde substitutes is motivated by two fundamental developments: the OSHA regulation standard declaring it hazardous and advocating its substitution with less dangerous chemicals and the fact that formalin is a poor preserver of nucleic acids. Among the non-alcoholic formalin substitute, glyoxal has been hailed as the best alternative. In this work, we showed that glyoxal-containing fixatives are not plausible polyvalent substitution options. PMID- 20005433 TI - [Sclerosing mesenteritis: can it be a IgG4 dysimmune disease?]. AB - AIMS: Mesenteric panniculitis (MP), sclerotic mesenteritis (SM) and mesenteric lipodystrophy (ML) are the three histopathological forms of a rare inflammatory mesenteric disease. The pathogenesis is unknown until now. Hypothesis of a dysimmune disorder is advocated. Our purpose was to find histological basis to assert this hypothesis. MATERIAL AND METHODS: All patients with a diagnosis of inflammatory mesenteric disease made in Beaujon hospital were selected. A histological study and a systematic immunostaining with IgG4, CD3 and CD8 antibodies were performed for each patient with paraffin block available. Dysimmune features were defined by: a plasma cells component >30 %, an eosinophilic polynuclears component >10 %, a rate TCD8/CD3 >30 % and IgG4+ plasma cells >30/high power field (x400). Cases were classified in 3 forms on the basis of the predominant component: MP (lymphoid/plasma cell infiltrate), SM (fibrosis), ML (fat necrosis). RESULTS: Thirteen patients (7 men, 6 women) were selected, with a mean age of 57 years (21-80 years); a paraffin block was available for 10 patients. The clinical and histological presentations were heterogeneous: a mesenteric mass was the most frequent mode of revelation (6/13 patients) and all histological forms were present (3 MP, 5 SM and 5 ML). Nine patients had surgical biopsy, 2 had a trephine biopsy and 2 a resection. Standard histological analysis showed numerous eosinophils (> or =10 %) and plasma cells (>30 %) in respectively 3 (23 %) and 6 (46 %) of the 13 cases. The immunohistochemistry (performed in 10 patients) showed numerous TCD8+ in 80 % of the cases and numerous IgG4+ plasma cells in only 31 % of the cases (4 cases) (IgG4+ >30/x400). CONCLUSIONS: Our results demonstrate that inflammatory mesenteric diseases are heterogeneous: high components of plasma cells, eosinophils and TCD8+ lymphocytes were found respectively in 46, 23 and 80 % of cases; eosinophils were observed exclusively in sclerotic forms. Only 31 % of cases encompass numerous IgG4+ plasma cells, suggesting a pathogenesis different than hyper-IgG4 diseases. PMID- 20005434 TI - [Immuno-histochemical study contribution in thoracic endometriosis: about an analysis of eight cases]. AB - Thoracic endometriosis (TE) is rare with positive histological diagnosis sometimes difficult, particularly in atypical form. The aim of this study was to identify features which can increase performance of the histolological TE diagnosis and more particularly immuno-histochemical (IHC) contribution with hormonal receptors, smooth muscle actin, Ber-Ep4, CD10 and calretinin antibodies. To address this issue, we retrieved, retrospectively, a large series of 591 pneumothorax operated. Among them, 135 (23%) were females including eight (6%) cases related to TE. Those eight women were surgically treated with resection of pleura (n=6/8), lung (n=5/8) and diaphragmatic samples (n=6/8). Typical histological lesions of endometriosis were observed in six cases among eight. All diaphragmatic samples presented, macroscopically, holes responsible of thoraco abdominal communication (n=6/6). Endometrial glands and/or endometrial stroma cells were found in the diaphragm (n=5/6) and in the pleura (n=2/6) but were never encountered in the lung (n=0/5). IHC study can confirm the five diaphragmatic localizations and can identify a new localization with expression of hormonal receptors, CD10 and smooth muscle actin in an island of fusiform cells. In conclusion, our study shows 1) the high frequency of diaphragmatic endometriosis localization which holes existence also can explain the pathogenesis, 2) the value of diaphragmatic samples in positive histological diagnosis of TE, 3) IHC interest to confirm endometriosis, particularly in atypical form and to differentiate from mesothelial cells inclusion. PMID- 20005435 TI - [Congenital pulmonary alveolar proteinosis related to a surfactant protein B deficiency: report of two cases]. AB - Congenital pulmonary alveolar proteinosis is an uncommon affection, distinct from adult's alveolar proteinosis by its clinical, pathological, etiological and evolutive characteristics. We report two cases of congenital alveolar proteinosis related to a surfactant protein B deficiency. Clinical presentations were similar: the two children were full-term newborns and had swiftly developed respiratory distress. Chest radiography demonstrated bilateral alveolar syndrome. Echocardiography was normal. There was no sign of infection. The two children died respectively at three weeks and two months of life. Lung biopsy showed lesions of alveolar proteinosis in the two cases. Both children were homozygotes for the 121ins2 mutation of the SFTPB gene. Diagnosis of surfactant protein B deficiency must be suspected in congenital alveolar proteinosis. It can be confirmed by the absence of detection of the surfactant B protein by immunohistochemistry on fixed and paraffin-embedded lung tissue or by western blot on bronchoalveolar fluid and by the absence of mRNA by RT-PCR. We report the value of molecular diagnosis for genetic counseling and the possibility of early prenatal diagnosis by trophoblast biopsy. PMID- 20005436 TI - [A rare congenital gingival tumor]. AB - INTRODUCTION: Congenital epulis (CE) of the newborn also known as congenital granular cell tumor or Neumann tumor is an uncommun benign tumour occuring in the anterior alveolar ridge of the jaws. It may interfer with breathing and feeding. In our study, we discuss the clinicopathologic and evolutive caracteristics and the diagnosis problems of this entity. OBSERVATION: A newborn girl, two months old, presented at birth a nodular mass in the upper alveolar crete of the maxilla. The mass measured 1.5 cm and exhibited a smooth surface. Histologically, it was composed of diffuse sheets and clusters of granular cells with abundant coarsely granular cytoplasm. The evolution was good without recurrence after five months. DISCUSSION: CE is a rare tumor; its etiopathogenesis is still unclear. The diagnosis suspected clinically is confirmed by the histopathological study. The prognosis is exellent without recurrence. PMID- 20005437 TI - [Isolated testicular mass revealing a metastasis of prostate adenocarcinoma]. AB - A testicular mass on a patient over 50 years should be considered as a metastasis, a lymphoma or a spermatocytic seminoma. Testicular metastases are rare, representing about 1 % of all testicular tumors. Prostate is the most common primary site, with less than 100 reported cases in the literature. They are most often incidentally discovered tumors, after bilateral orchiectomy for hormonal management of prostate carcinoma or at autopsy. Metastases revealed by a testicular mass are very rare. We report the original case of an 80-year-old patient with a left testicular mass indicating a metastasis of a prostate carcinoma diagnosed six years earlier. No other metastatic localization was found. PMID- 20005438 TI - [Retiform haemangioendothelioma: a case report]. AB - Retiform haemangioendothelioma is a locally aggressive, very rarely metastasizing vascular lesion. Histologically, it is characterized by distinctive arborizing blood vessels resembling "rete testis" and lined by endothelial cells with characteristic hobnail morphology. We present an additional case, in the leg of a 64-year-old patient. We discuss the classification of hemangioendotheliomas. The term hemangioendothelioma should be restricted to vascular tumours of "intermediate malignancy" but has been used to designate tumours with variable histological features and clinical behaviour. Spindle cell hemangio(endothelio)ma is currently regarded as a benign reactive lesion. Kaposiform hemangioendothelioma is potentially lethal due to consumption coagulopathy but no metastasizing case has been reported. Epithelioid hemangioendothelioma is associated with a significant metastatic risk and has been included in the category of malignant vascular tumors. The vascular lesions fulfilling the strict definition of hemangioendothelioma include retiform hemangioendothelioma, papillary intralymphatic angioendothelioma "Dabska's tumor", composite hemangioendothelioma and perhaps the controversial polymorphic hemangioendothelioma. PMID- 20005439 TI - [Muir-Torre syndrome: rare association with duodenal carcinoma]. AB - Muir-Torre syndrome, a rare autosomal dominant inherited disease, is characterized by the synchronous or metachronous occurrence of at least one sebaceous gland neoplasia such as an adenoma or carcinoma, with or without keratoacanthoma, and at least one internal malignancy, mostly colorectal cancer. Visceral malignant neoplasms seem to be less aggressive than their sporadic counterparts. Muir-Torre syndrome has been recognised as a subset of Lynch's syndrome, with similar microsatellite instability and germline mutations in DNA mismatch repair (MMR) genes mainly in MSH2 and/or MLH1. We report the case of a 60-year-old man with a Muir-Torre syndrome, presenting an indolent poorly differentiated duodenal carcinoma. Immunohistochemical analysis revealed the loss of expression of MSH2 and MSH6 proteins in tumor cells. According to medical literature, only 16 cases of Muir-Torre syndrome with small bowel carcinoma have been reported to date. PMID- 20005440 TI - [Inflammatory fibroid polyp: a report of three cases including an oesophageal tumor]. AB - We report three different cases in location with an original occurrence in the oesophagus. The gross appearance showed polypoid pedonculated masses with a whitish cut surface, arising from the submucosa. On microscopic examination, the tumor consisted of spindle cell dispersed in a loose fibromyxoid stroma containing numerous blood vessels and inflammatory cells with abundant eosinophils. At immunohistochemistry, the cells were strongly positive for CD34. Inflammatory fibroid polyp is a rare benign lesion of the gastro-intestinal tract. The pathogenesis of this lesion remains throat and unclear. Diagnosis on biopsy is difficult with various histologic pattern and because the inflammatory fibroid polyp share common pathologic features with digestive spindle cells tumor. PMID- 20005441 TI - [The presence of mycobacteria in bronchoalveolar lavage fluid from an immunocompetent patient does not necessarily imply tuberculosis]. AB - Mycobacterium tuberculosis is the most frequently identified mycobacterium in the bronchoalveolar lavage fluid (BALF) of immunocompetent patients. Lung infections due to non-tuberculous mycobacteria (NTM) are rare in such patients and then often occur in the context of pre-existing chronic lung disease. We report the case of an immunocompetent 85-year-old woman without pre-existing lung disease in whom M. abscessus was recovered from BALF. Cytological examination of the BALF revealed an increased number of neutrophils and some acid-fast bacilli, all located within neutrophil cytoplasm. This case report contributes a cytological description of BALF in the context of M. abscessus infection, which is poorly detailed in the literature. PMID- 20005442 TI - [Tumor-to-tumor metastasis: report of three cases]. AB - Tumor-to-tumor metastasis is a very rare event. We report three cases of tumor metastasizing in another tumor: a clear cell renal cell carcinoma in a vesicular thyroid adenoma, a lung carcinoma in a meningioma and a neuroendocrine lung carcinoma in a clear cell renal cell carcinoma. According to the literature, clear cell renal cell carcinoma is the most common tumor recipient of metastasis while lung carcinoma is the most common donor tumor. Several physiopathological mechanisms can explain this phenomenon, but many of them are still unknown. PMID- 20005443 TI - [A tumor of the spleen]. PMID- 20005444 TI - [A particular tumour of the shoulder]. PMID- 20005445 TI - [A pineal tumour in an infant]. PMID- 20005446 TI - [Fatal cardiac failure following antibiotic treatment]. PMID- 20005447 TI - [An unusual mutinodular goiter]. PMID- 20005448 TI - [Duodenal biopsy for chronic diarrhea]. PMID- 20005449 TI - [Chronic lymphocytic leukemia may be revealed by prostatic symptoms]. PMID- 20005450 TI - Quality measures, standards, and accreditation for breast centers in the United States. PMID- 20005451 TI - Lin28-let7 modulates radiosensitivity of human cancer cells with activation of K Ras. AB - PURPOSE: To evaluate the potential of targeting Lin28-let7 microRNA regulatory network for overcoming the radioresistance of cancer cells having activated K-Ras signaling. METHODS AND MATERIALS: A549 lung carcinoma cells and ASPC1 pancreatic cancer cells possessing K-RAS mutation were transfected with pre-let7a microRNA or Lin28 siRNA, respectively. Clonogenic assay, quantitative reverse transcription polymerase chain reaction, and Western analysis were performed. The effects of Lin28 on SQ20B cells having wild-type K-RAS, and a normal fibroblast were also assessed. RESULTS: The overexpression of let-7a decreased expression of K-Ras and radiosensitized A549 cells. Inhibition of Lin28, a repressor of let-7, attenuated K-Ras expression and radiosensitized A549 and ASPC1 cells. Neither SQ20B cells expressing wild-type K-RAS nor HDF, the normal human fibroblasts, were radiosensitized by this approach. CONCLUSIONS: The Lin28-let7 regulatory network may be a potentially useful therapeutic target for overcoming the radioresistance of human cancers having activated K-Ras signaling. PMID- 20005452 TI - Microsatellite instability predicts clinical outcome in radiation-treated endometrioid endometrial cancer. AB - PURPOSE: To elucidate whether microsatellite instability (MSI) predicts clinical outcome in radiation-treated endometrioid endometrial cancer (EEC). METHODS AND MATERIALS: A consecutive series of 93 patients with EEC treated with extrafascial hysterectomy and postoperative radiotherapy was studied. The median clinical follow-up of patients was 138 months, with a maximum of 232 months. Five quasimonomorphic mononucleotide markers (BAT-25, BAT-26, NR21, NR24, and NR27) were used for MSI classification. RESULTS: Twenty-five patients (22%) were classified as MSI. Both in the whole series and in early stages (I and II), univariate analysis showed a significant association between MSI and poorer 10 year local disease-free survival, disease-free survival, and cancer-specific survival. In multivariate analysis, MSI was excluded from the final regression model in the whole series, but in early stages MSI provided additional significant predictive information independent of traditional prognostic and predictive factors (age, stage, grade, and vascular invasion) for disease-free survival (hazard ratio [HR] 3.25, 95% confidence interval [CI] 1.01-10.49; p = 0.048) and cancer-specific survival (HR 4.20, 95% CI 1.23-14.35; p = 0.022) and was marginally significant for local disease-free survival (HR 3.54, 95% CI 0.93 13.46; p = 0.064). CONCLUSIONS: These results suggest that MSI may predict radiotherapy response in early-stage EEC. PMID- 20005453 TI - Outcomes following iodine-125 monotherapy for localized prostate cancer: the results of leeds 10-year single-center brachytherapy experience. AB - PURPOSE: This study reports the 10-year experience of permanent brachytherapy monotherapy at a single UK center. METHODS AND MATERIALS: Between March 1995 and September 2004, 1,298 patients underwent trans-rectal ultrasound (TRUS) planned transperineal brachytherapy delivering 145 Gy using I-125. No patient received supplemental external beam; 44.2% received neoadjuvant hormones. In 688, CT postimplant dosimetry was available. Outcome data were analyzed in terms of overall survival (OS), disease specific survival (DSS), and PSA relapse-free survival (PSA-RFS). RESULTS: The mean age was 62.9 (range, 34-83) years. Median follow-up was 4.9 years (range, 2.03-11.7 years). OS and DSS were 85% and 95%, respectively, at 10 years. Twenty-one patients died from prostate cancer (1.6%) and 34 (2.5%) from unrelated causes. Seventy-four (5.7%) developed evidence of clinical failure. Overall PSA-RFS was 79.9% and 72.1% at 10 years (American Society for Therapeutic Radiology and Oncology [ASTRO] and Nadir+2 definitions, respectively). Higher presenting PSA or Gleason score and use of neoadjuvant hormones were associated with an increased risk of biochemical failure (p <0.01). Biochemical control was achieved in 86.4%, 76.7%, and 60.6% (ASTRO) and 72.3%, 73.5%, and 57.6% (Nadir+2) of patients in low-, intermediate-, and high-risk groups, respectively. Biochemical control was achieved in 88% of patients with D(90) > or =140 Gy and in 78% of patients with D(90) <140 Gy (p <0.01). CONCLUSIONS: I-125 brachytherapy alone achieved excellent rates of medium-term biochemical control in both low- and selected intermediate-risk localized prostate cancer patients. Postimplant dosimetry improved with experience and longer follow-up, confirming the relationship of D(90) with biochemical control. PMID- 20005454 TI - Distinct signaling pathways after higher or lower doses of radiation in three closely related human lymphoblast cell lines. AB - PURPOSE: The tumor suppressor p53 plays an essential role in cellular responses to DNA damage caused by ionizing radiation; therefore, this study aims to further explore the role that p53 plays at different doses of radiation. MATERIALS AND METHODS: The global cellular responses to higher-dose (10 Gy) and lower dose (iso survival dose, i.e., the respective D0 levels) radiation were analyzed using microarrays in three human lymphoblast cell lines with different p53 status: TK6 (wild-type p53), NH32 (p53-null), and WTK1 (mutant p53). Total RNAs were extracted from cells harvested at 0, 1, 3, 6, 9, and 24 h after higher and lower dose radiation exposures. Template-based clustering, hierarchical clustering, and principle component analysis were applied to examine the transcriptional profiles. RESULTS: Differential expression profiles between 10 Gy and iso survival radiation in cells with different p53 status were observed. Moreover, distinct gene expression patterns were exhibited among these three cells after 10 Gy radiation treatment, but similar transcriptional responses were observed in TK6 and NH32 cells treated with iso-survival radiation. CONCLUSIONS: After 10 Gy radiation exposure, the p53 signaling pathway played an important role in TK6, whereas the NFkB signaling pathway appeared to replace the role of p53 in WTK1. In contrast, after iso-survival radiation treatment, E2F4 seemed to play a dominant role independent of p53 status. This study dissected the impacts of p53, NFkB and E2F4 in response to higher or lower doses of gamma-irradiation. PMID- 20005455 TI - Role of intercellular adhesion molecule-1 in radiation-induced brain injury. AB - PURPOSE: To determine the role of intercellular adhesion molecule-1 (ICAM-1) in the pathogenesis of brain injury after irradiation (IR). METHODS AND MATERIALS: We assessed the expression of ICAM-1 in mouse brain after cranial IR and determined the histopathologic and behavioral changes in mice that were either wildtype (+/+) or knockout (-/-) of the ICAM-1 gene after IR. RESULTS: There was an early dose-dependent increase in ICAM-1 mRNA and protein expression after IR. Increased ICAM-1 immunoreactivity was observed in endothelia and glia of ICAM 1+/+ mice up to 8 months after IR. ICAM-1-/- mice showed no expression. ICAM-1+/+ and ICAM-1-/- mice showed similar vascular abnormalities at 2 months after 10-17 Gy, and there was evidence for demyelination and inhibition of hippocampal neurogenesis at 8 months after 10 Gy. After 10 Gy, irradiated ICAM-1+/+ and ICAM 1-/- mice showed similar behavioral changes at 2-6 months in open field, light dark chamber, and T-maze compared with age-matched genotype controls. CONCLUSION: There is early and late upregulation of ICAM-1 in the vasculature and glia of mouse brain after IR. ICAM-1, however, does not have a causative role in the histopathologic injury and behavioral dysfunction after moderate single doses of cranial IR. PMID- 20005456 TI - Irradiation with carbon ion beams induces apoptosis, autophagy, and cellular senescence in a human glioma-derived cell line. AB - PURPOSE: We examined biological responses of human glioma cells to irradiation with carbon ion beams (C-ions). METHODS AND MATERIALS: A human glioma-derived cell line, NP-2, was irradiated with C-ions. Apoptotic cell nuclei were stained with Hoechst 33342. Induction of autophagy was examined either by staining cells with monodansylcadaverine (MDC) or by Western blotting to detect conversion of microtuble-associated protein light chain 3 (MAP-LC3) (LC3-I) to the membrane bound form (LC3-II). Cellular senescence markers including induction of senescence-associated beta-galactosidase (SA-beta-gal) were examined. The mean telomere length of irradiated cells was determined by Southern blot hybridization. Expression of tumor suppressor p53 and cyclin/cyclin-dependent kinase inhibitor p21(WAF1/CIP1) in the irradiated cells was analyzed by Western blotting. RESULTS: When NP-2 cells were irradiated with C-ions at 6 Gy, the major population of the cells died of apoptosis and autophagy. The residual fraction of attached cells (<1% of initially irradiated cells) could not form a colony: however, they showed a morphological phenotype consistent with cellular senescence, that is, enlarged and flattened appearance. The senescent nature of these attached cells was further indicated by staining for SA-beta-gal. The mean telomere length was not changed after irradiation with C-ions. Phosphorylation of p53 at serine 15 as well as the expression of p21(WAF1/CIP1) was induced in NP-2 cells after irradiation. Furthermore, we found that irradiation with C-ions induced cellular senescence in a human glioma cell line lacking functional p53. CONCLUSIONS: Irradiation with C-ions induced apoptosis, autophagy, and cellular senescence in human glioma cells. PMID- 20005457 TI - A murine model for the study of molecular pathogenesis of radiation proctitis. AB - PURPOSE: To establish a novel mouse brachytherapy model with which to study the role of inflammation in the pathogenesis of radiation proctitis. METHODS AND MATERIALS: The distal rectums of BALB/c and C57BL/6 mice were irradiated with three to five fractions of 5.5 to 8 Gy. Tissues were harvested and evaluated for histopathology, using the radiation injury score (RIS). Cytokine mRNA expression was assessed using real-time PCR. RESULTS: Fifty percent of the mice treated with 22 Gy delivered in four fractions of 5.5 Gy died as a result of anorectal stenosis and distal bowel obstruction prior to the time of scheduled sacrifice, with a latency period of 4 to 10 weeks for the BALB/c and 3 to 4 weeks for the C57BL/6 mice. The RISs were 7, 12, and 8 at 2, 6, and 11 weeks, respectively, in the BALB/c mice and was 8.7 in the C57BL/6 mice on week 6. A 100- to 300-fold increase in interleukin-1beta (IL-1beta) (p = 0.04) and IL-6 mRNA (p = 0.07) and a 5- to 6-fold increase in transforming growth factor (TGF) and tumor necrosis factor-alpha mRNA expression levels (p < 0.001 and p = 0.01) were observed at 2 to 6 weeks after radiation. Cytokine mRNA tissue expression correlated positively with radiation dose (p < 0.0001). The RIS correlated well with IL-1beta and IL-6 mRNA levels in the BALB/c mice and with IL-1beta, IL-6, and TGF mRNA levels in C57BL/6 mice. Analysis of receiver operating characteristic curve showed that IL 1beta and IL-6 have the largest area under the curve and therefore are good markers of radiation proctitis (p < 0.001). CONCLUSIONS: Radiation-induced proctitis was associated with a dose-dependent, characteristic proinflammatory cytokine response pattern in a novel mouse model suitable for interventional studies. PMID- 20005458 TI - Imaging tumor variation in response to photodynamic therapy in pancreatic cancer xenograft models. AB - PURPOSE: A treatment monitoring study investigated the differential effects of orthotopic pancreatic cancer models in response to interstitial photodynamic therapy (PDT), and the validity of using magnetic resonance imaging as a surrogate measure of response was assessed. METHODS AND MATERIALS: Different orthotopic pancreatic cancer xenograft models (AsPC-1 and Panc-1) were used to represent the range of pathophysiology observed in human beings. Identical dose escalation studies (10, 20, and 40J/cm) using interstitial verteporfin PDT were performed, and magnetic resonance imaging with T2-weighted and T1-weighted contrast were used to monitor the total tumor volume and the vascular perfusion volume, respectively. RESULTS: There was a significant amount of necrosis in the slower-growing Panc-1 tumor using high light dose, although complete necrosis was not observed. Lower doses were required for the same level of tumor kill in the faster-growing AsPC-1 cell line. CONCLUSIONS: The tumor growth rate and vascular pattern of the tumor affect the optimal PDT treatment regimen, with faster growing tumors being relatively easier to treat. This highlights the fact that therapy in human beings shows a heterogeneous range of outcomes, and suggests a need for careful individualized treatment outcomes assessment in clinical work. PMID- 20005459 TI - The development and clinical use of a beam ON-LINE PET system mounted on a rotating gantry port in proton therapy. AB - PURPOSE: To verify the usefulness of our developed beam ON-LINE positron emission tomography (PET) system mounted on a rotating gantry port (BOLPs-RGp) for dose volume delivery-guided proton therapy (DGPT). METHODS AND MATERIALS: In the proton treatment room at our facility, a BOLPs-RGp was constructed so that a planar PET apparatus could be mounted with its field of view covering the iso center of the beam irradiation system. Activity measurements were performed in 48 patients with tumors of the head and neck, liver, lungs, prostate, and brain. The position and intensity of the activity were measured using the BOLPs-RGp during the 200 s immediately after the proton irradiation. RESULTS: The daily measured activity images acquired by the BOLPs-RGp showed the proton irradiation volume in each patient. Changes in the proton-irradiated volume were indicated by differences between a reference activity image (taken at the first treatment) and the daily activity-images. In the case of head-and-neck treatment, the activity distribution changed in the areas where partial tumor reduction was observed. In the case of liver treatment, it was observed that the washout effect in necrotic tumor cells was slower than in non-necrotic tumor cells. CONCLUSIONS: The BOLPs RGp was developed for the DGPT. The accuracy of proton treatment was evaluated by measuring changes of daily measured activity. Information about the positron emitting nuclei generated during proton irradiation can be used as a basis for ensuring the high accuracy of irradiation in proton treatment. PMID- 20005461 TI - Response to "Role of external beam radiotherapy in patients with advanced or recurrent nonanaplastic thyroid cancer: Memorial Sloan-Kettering Cancer Center experience" (Int J Radiat Oncol Biol Phys 2009;73:795-801). PMID- 20005463 TI - Clinical experience with image-guided radiotherapy in an accelerated partial breast intensity-modulated radiotherapy protocol: in regard to Leonard CE et al. (Int J Radiat Oncol Biol Phys Epub May 19, 2009). PMID- 20005464 TI - Relationship between serum protein and mortality in adults on long-term hemodialysis: exhaustive review and meta-analysis. AB - The aim of this exhaustive review and meta-analysis was to explore the relation among serum protein, inflammatory markers, and all-cause and cardiovascular mortalities in adult patients on maintenance hemodialysis. We searched the Medline, Science Citation Index, Academic Search Premier, Cochrane Library, and Embase electronic data bases. Data extraction and quality assessment were done independently by two reviewers and results were pooled using the random effects model. Cochran's Q was used to identify heterogeneity and a funnel plot was used for assessment of publication bias. A meta-analysis was performed on 38 studies (265 330 patients) reporting on serum proteins, inflammatory markers, and mortality. A significant inverse relation was found between serum albumin and all cause (hazard ratio [HR] 0.7038, 95% confidence interval [CI] 0.6367-0.7781) and cardiovascular (HR 0.8726, 95% CI 0.7909-0.9628) mortalities, with a significantly stronger relation with all-cause mortality (P=0.0014). Pooled results for C-reactive protein showed a weak but significant direct relation with all-cause mortality (HR 1.0322, 95% CI 1.0151-1.0496), but there was not a significant relation between C-reactive protein and cardiovascular mortality (HR 1.0172, 95% CI 0.9726-1.0639). A high degree of heterogeneity was identified among studies especially in the case of all-cause mortality. An asymmetrical funnel plot for serum albumin is suggestive of publication bias. From the meta analysis it is concluded that serum albumin showed a significant inverse relation with all-cause and cardiovascular mortalities but the relation between prealbumin and all-cause mortality was not significant. C-reactive protein showed a significant direct relation with all-cause mortality but not with cardiovascular mortality. The potential adverse effects of malnutrition and infections in relation to mortality highlight the need for continued treatment of infections and correction of malnutrition in patients on dialysis. PMID- 20005465 TI - Inhaled insulin-intrapulmonary, intranasal, and other routes of administration: mechanisms of action. AB - BACKGROUND: After discovery of insulin as a hypoglycemic agent in 1921 various routes of administration to control blood glucose were attempted. These included subcutaneous, oral, rectal, sublingual, buccal, transdermal, vaginal, intramuscular, intrapulmonary and intranasal delivery systems. While each delivery system controlled hyperglycemia the subcutaneous route was given priority until 2006 when the Federal Drug Administration (FDA) approved the first commercially available pulmonary inhaled insulin. METHODS: A review of major publications dealing with intrapulmonary administration of insulin was made to understand the physiological basis for its use, its efficacy in controlling hyperglycemia, its side effects and a comparison of its efficacy with other delivery methods. RESULTS: The large surface area of the lung, its good vascularization, capacity for solute exchange and ultra thin membranes of alveolar epithelia are unique features that facilitate pulmonary insulin delivery. Large lung surface area ( approximately 75 m(2)) and thin alveolar epithelium ( approximately 0.1-0.5 microm) permit rapid drug absorption. First pass metabolism avoids gastrointestinal tract metabolism. Lung drug delivery depends upon a complex of factors including size, shape, density, charge and pH of delivery entity, velocity of entry, quality of aerosol deposition, character of alveoli, binding characteristics of aerosol on the alveolar surface, quality of alveolar capillary bed and its subsequent vascular tree. Many studies were performed to optimize each of these factors using several delivery systems to enhance pulmonary absorption. Availability was about 80% of subcutaneous administration with peak activity within 40-60 min of administration. Intranasal insulin delivery faces a smaller surface area ( approximately 180 cm(2)) with quite different absorption characteristics in nasal epithelium and its associated vasculature. Absorption depends upon many factors including composition and character of nasal mucus. Absorption of intranasal insulin resulted in a faster absorption time course than with subcutaneous insulin. INTERPRETATION: After many studies the FDA approved Pfizer's product, Exubera, for intrapulmonary insulin delivery. While the system was effective its expense and putative side effects caused the drug company to withdraw the drug from the marketplace. Attempts by other pharmaceutical companies to use intrapulmonary insulin delivery are presently being made as well as some minor attempts to use intranasal delivery systems. PMID- 20005466 TI - Stunting, adiposity, and low-grade inflammation in African adolescents from a township high school. AB - OBJECTIVE: The objective of this study was to compare the inflammatory status of children with differences in nutritional status. METHODS: This was a cross sectional study of 184 African children aged 13-18 y from a low socioeconomic background that compared stunted with non-stunted and lean with over-fat (percentage of body fat above normal cutoff points) children. Fasting serum tumor necrosis factor-alpha, interleukin-6, C-reactive protein, and insulin were measured using high-sensitivity methods. Body composition was assessed using anthropometry and air-displacement plethysmography. T tests for parametric data and the Mann-Whitney test for non-parametric data were used to compare groups. Regression analyses and principal components analyses were done to assess relations between body composition and biochemical variables. RESULTS: Of all participants 18% were stunted. Serum tumor necrosis factor-alpha of stunted girls was higher than in non-stunted girls. More of the stunted boys were over-fat compared with their non-stunted counterparts. Regression analyses showed that insulin resistance, diastolic blood pressure, and C-reactive protein contributed significantly to interleukin-6 in boys. Serum C-reactive protein, waist circumference, and body mass index clustered together in factor analysis in boys. Serum interleukin-6, waist-hip ratio, and tumor necrosis factor-alpha clustered together in factor analysis in girls. CONCLUSION: An association between adiposity and stunting and between adiposity low-grade inflammation was found in this study. Interventions for stunted children focus mainly on correction of undernutrition by providing feeding schemes. Attention should, however, also be paid to changes in body composition over time to prevent excessive abdominal fat accumulation and risk for cardiovascular diseases later in life. PMID- 20005467 TI - Difructose anhydride III promotes iron absorption in the rat large intestine. AB - OBJECTIVES: We examined the intestinal segment responsible for and the mechanisms involved in the non-digestible disaccharide difructose anhydride III (DFAIII) mediated enhancement of iron absorption. METHODS: Net iron and calcium absorption (percentage) were observed in male Sprague-Dawley rats fed an AIN93G-based diet with or without DFAIII (30 g/kg of diet) for 2 wk after cecectomy or laparotomy (experiment 1). The absorption rates of calcium and iron from ligated jejunal loops (experiment 2) or ligated cecal sacs (experiment 3) in anesthetized rats fed a control or DFA diet were observed with or without the addition of DFAIII to the instilled mucosal fluid. Levels of the major iron transporter divalent metal transporter-1 (DMT-1) mRNA per 28S rRNA were evaluated by real-time polymerase chain reaction in the intestinal mucosa. RESULTS: Net absorption (percentage) of iron and calcium was higher in the DFA group than in the DFA-free group in laparotomized rats. The enhancement of iron absorption was almost and calcium absorption was partly abolished by cecectomy. The absorption rate of calcium, but not of iron, from the jejunal loops was increased by the addition of DFAIII (100 mmol/L). Iron absorption was clearly higher in the ligated cecal sacs of the DFAIII-fed group than in the DFA-free group with or without the mucosal addition of DFAIII. DMT-1 mRNA level per 28S rRNA was maintained in the cecal mucosa through large increases in cecal size due to DFAIII feeding. CONCLUSION: The DFAIII-induced increases in iron absorption are the result of increased cecal iron absorptive capacity through expansion of the cecal mucosa maintaining DMT-1 mRNA expression. PMID- 20005468 TI - Combined inhalational and oral supplementation of ascorbic acid may prevent influenza pandemic emergency: a hypothesis. AB - Occurrence of influenza pandemics is a worldwide phenomenon and a significant cause of mortality and morbidity throughout the globe. It is due to mutations in the influenza virus genetic material creating antigenic drift of pathogenic viral proteins resulting in emergence of new influenza virus strains. Therefore, the vaccines available for prevention of influenza offer no protection against influenza pandemics caused by new virus strains. Moreover, the existing drugs used to combat influenza may be ineffective to treat influenza pandemics due to the emergence of drug resistance in the pandemic virus strain. Therefore, a working strategy must be developed to combat influenza pandemics. In this review we have addressed this problem and reviewed the published studies on ascorbic acid in the common cold and influenza and laboratory studies on the effect of ascorbic acid on influenza virus. We have also correlated the clinical and laboratory studies and developed a hypothesis to prevent influenza pandemics. PMID- 20005470 TI - Comment on "Association of some specific nutrient deficiencies with periodontal disease in elderly people: a systematic literature review". PMID- 20005469 TI - Coronary artery bypass graft surgery depletes plasma thiamine levels. AB - OBJECTIVE: Thiamine is an essential component of cellular metabolism, and lack of this vitamin results in a potentially life-threatening biochemical lesion. The stress of surgery and critical disease depletes electrolytes, minerals, and essential biochemical substrates. We hypothesized that critical illness (represented by major surgery) would result in decreased thiamine levels over time. METHODS: We performed a prospective, observational study of serial thiamine levels of 15 patients who underwent non-emergent coronary artery bypass graft surgery. The primary endpoint was change in thiamine levels from before to immediately after surgery. Secondary endpoints included change in thiamine levels from presurgical to 6- and 24-h time points. RESULTS: Of the 15 study patients, 1 did not have a plasma thiamine measurement at time 0 because of laboratory error and could not be accounted for in paired comparisons over time. Plasma thiamine levels decreased significantly from before to after coronary artery bypass grafting (P=0.0004). In addition, there was a statistically significant decrease in thiamine levels from before surgery to 24h (P=0.003). CONCLUSION: Our data suggest that major surgery (as a surrogate for the stress of critical illness) depletes thiamine levels; further study is needed to determine whether routine replacement of thiamine in the critically ill is warranted. PMID- 20005473 TI - Biomarkers in peripheral arterial disease. AB - Patients who have peripheral arterial disease (PAD) have a high incidence of cardiac morbidity and mortality. There have been numerous biomarkers described to assess cardiovascular risk, but few are part of routine clinical practice. Currently, the key biomarkers that improve risk stratification in patients with PAD are cardiac troponins, C-reactive protein, and B-type natriuretic peptide. Recent advances in descriptive proteomics will offer future potential for biomarker discovery. However, it is essential that new markers are translated into tools for patient care. This review examines the potential biomarkers that improve cardiovascular risk stratification in PAD and avenues for future studies. PMID- 20005474 TI - Heart rate reduction by I(f)-channel inhibition and its potential role in heart failure with reduced and preserved ejection fraction. AB - Selective heart rate (HR) reduction by I(f)-channel inhibition is a recently developed pharmacological principle in cardiovascular therapy. Among these newly identified HR-lowering drugs, only ivabradine has now become approved for clinical use. I(f)-channel inhibition mainly reduces HR, thereby improving myocardial oxygen supply, energy balance, and cardiac function. Ivabradine was well tolerated and revealed a good safety profile in the investigated study populations. The guiding experimental and clinical results of I(f)-channel inhibition were compared to those of beta-blockade as a HR reducing principle as well as cornerstone of heart failure standard therapy. Beside its use in therapy of coronary artery disease, I(f)-channel inhibition potentially exhibits beneficial effects in systolic and diastolic heart failure as well. Therefore, hemodynamic effects of ivabradine and its limitations in heart failure together with the biological impact of HR reduction will be considered in this context. Because no clinical data with specific heart-rate-reducing agents are available in heart failure patients until now, the prospective significance of I(f)-channel inhibition can only be speculated on. However, the presented results and considerations are encouraging: ivabradine may play a therapeutic role in the future protecting left ventricular function and structure from early deterioration in heart failure with reduced and preserved ventricular ejection fraction. PMID- 20005476 TI - Galectins in atherosclerotic disease. AB - Although cells constantly adapt to external humoral cues, like cytokines and hormones, in practice, most cellular behavior is under locally guided control, via cell-cell interactions. The most prominent members of this group of molecules involved in this intercellular signaling are the galectins, a family of carbohydrate-binding proteins, which by binding specific receptors induce signaling. It has become clear that galectins are important in the pathogenesis of cardiovascular disease, in particular atherosclerosis, stroke, myocardial infarction, and heart failure. Modulation of galectin signaling holds great promise for the treatment of cardiovascular disease as evident from preclinical studies. PMID- 20005475 TI - Aldehyde dehydrogenase 2 in cardiac protection: a new therapeutic target? AB - Mitochondrial aldehyde dehydrogenase 2 (ALDH2) is emerging as a key enzyme involved in cytoprotection in the heart. ALDH2 mediates both the detoxification of reactive aldehydes such as acetaldehyde and 4-hydroxy-2-nonenal and the bioactivation of nitroglycerin to nitric oxide. In addition, chronic nitrate treatment results in ALDH2 inhibition and contributes to nitrate tolerance. Our laboratory recently identified ALDH2 to be a key mediator of endogenous cytoprotection. We reported that ALDH2 is phosphorylated and activated by the survival kinase protein kinase C epsilon and found a strong inverse correlation between ALDH2 activity and infarct size. We also identified a small molecule ALDH2 activator which reduces myocardial infarct size induced by ischemia/reperfusion in vivo. In this review, we discuss evidence that ALDH2 is a key mediator of endogenous survival signaling in the heart, suggest possible cardioprotective mechanisms mediated by ALDH2 and discuss potential clinical implications of these findings. PMID- 20005477 TI - The role of notch signaling in endothelial progenitor cell biology. AB - It is generally accepted that endothelial progenitor cells (EPCs) can promote postnatal neovascularization and be used for vascular regeneration, thus representing a promising new tool for the treatment of cardiovascular diseases. However, the exact molecular mechanisms and signaling pathways regulating the proliferation, differentiation, and migration of EPCs; their interaction with niche cells; and their regenerative capacity still remain elusive. The Notch signaling pathway shown to be important for the maintenance and differentiation of various stem and progenitor populations is also involved in EPC regulation. In this review, we will summarize the current knowledge about the pivotal role of Notch signaling in EPC biology and EPC-mediated vascular regeneration. PMID- 20005479 TI - Management of renal transplant complications and renal artery embolization for nononcology etiologies. Introduction. PMID- 20005478 TI - The role of HMGCR alternative splicing in statin efficacy. AB - Statins, or 3-hydroxy-3-methylglutaryl coenzyme A reductase (HMGCR) inhibitors, are widely prescribed to lower plasma cholesterol levels and reduce cardiovascular disease risk. Despite the well-documented efficacy of statins, there is large interindividual variation in response. Using a panel of immortalized lymphocyte cell lines incubated with simvastatin, we recently found that the magnitude of expression of an alternatively spliced HMGCR transcript lacking exon 13 was inversely correlated with in vivo reductions of total cholesterol, low-density lipoprotein cholesterol, apoB, and triglycerides after statin treatment of the individuals from whom the cells were derived. This review will discuss the potential significance of alternative splicing as a mechanism contributing to variation in statin efficacy as well as the use of immortalized lymphocyte cell lines for identifying pharmacogenetically relevant polymorphisms and molecular mechanisms. PMID- 20005480 TI - Transcatheter renal artery embolization: clinical applications and techniques. AB - Renal artery embolization is a minimally invasive procedure that is increasingly being used for treatment of a wide range of conditions. The main indications for renal artery embolization include (1) prenephrectomy and preradiofrequency ablation infarction of renal tumors, (2) management of renal angiomyolipomas, (3) palliations of unresectable renal malignancy, (4) renal hemorrhage (life threatening or chronic debilitating hematuria), (5) arteriovenous fistulas, (6) vascular malformations, (7) renal artery aneurysms and pseudoaneurysms, and other less common indications, such as severe hydronephrosis and hypertension. A variety of embolic materials are available, such as metal coils, sclerosants (glue, Onyx, absolute ethanol, lipiodol), and particulate embolic agents (polyvinyl alcohol particles and embospheres). Selection of the appropriate agent depends on the clinical application, technical and clinical endpoints, as well as the pathology lesion(s) targeted. Renal artery embolization can be performed alone or in combination with remodeling techniques, stent-grafting, and balloon- or stent-assisted coiling in more complicated cases. The procedure is generally regarded as safe and effective for diverse applications and is considered as an evolving area in the field of endoluminal therapy. PMID- 20005481 TI - Management of vascular and nonvascular complications after renal transplantation. AB - Renal transplantation is the treatment of choice for end-stage renal disease. Despite medical and surgical advances, vascular and nonvascular complications remain common post transplantation, occurring in 12%-20% of patients (Kobayashi K, Censullo ML, Rossman LL, et al: Radiographics 27:1109-1130, 2007; Orons PD, Zajko AB: Radiol Clin North Am 33:461-471, 1995). Complications of renal transplantation can range from minor complications, such as peri-graft fluid collections, to severe complications, such as renal vein thrombosis or transplant renal artery stenosis (TRAS). These complications may compromise graft function and cause significant morbidity. Most postoperative complications can be diagnosed by radiologic evaluation and often times can be treated by minimally invasive, interventional radiologic procedures. A thorough understanding of how the complications impair allograft function and survival is essential in allowing adequate treatment. Interventional radiology plays an invaluable role in the postoperative management of renal transplantation and related complications. The general indications for renal transplantation related procedures and the most commonly used and latest techniques are described in more detail. PMID- 20005482 TI - The HIV-associated nephrologist: Advice straight from the HAART. PMID- 20005483 TI - HIV through a nephrologist's lens. PMID- 20005484 TI - What does the generalist need to know about HIV infection? AB - Despite recent improvements in the efficacy, safety, tolerability, and convenience of antiretroviral therapy for patients, the management of HIV infection remains complex for clinicians. Multiple studies have shown better clinical outcomes and lower cost of care when HIV-infected patients are managed by experts. However, generalists are frequently involved in the care of patients with HIV infection, in many cases providing primary care in collaboration with an HIV expert. Generalists also play a critical role in the diagnosis and prevention of HIV infection. Generalists managing HIV-infected patients should be aware of the components of the initial patient evaluation. They should be familiar with the general principles of antiretroviral therapy and opportunistic infection prevention. They should be able to recognize antiretroviral toxicity and should be aware of common drug-drug interactions involving antiretroviral agents. PMID- 20005485 TI - HIV and CKD epidemiology. AB - Nephrologists can serve many important functions for HIV-infected patients, including identifying risks for developing kidney disease, detecting and diagnosing kidney disease, distinguishing antiretroviral-induced kidney injury from kidney disease in the setting of antiretroviral therapy, comanaging the clinical course and complications of CKD, and preparing patients for dialysis and/or transplantation. The epidemiology of kidney disease in HIV informs us for these functions by describing the natural history of disease, its frequent occurrence in high-risk communities, and its potential causes. Risk factors that drive CKD in HIV are black race, hypertension, diabetes, HIV viral replication with low CD4 cell counts, high viral load or acquired immune deficiency syndrome defining conditions, and antiretroviral agents with nephrotoxic potential. The prevalence of these risk factors in any population determines the magnitude of the problem, which can range from as low as 2% to as high as 30%. Recent research focuses on kidney health in HIV. Important links between HIV viral replication and glomerular filtration rate, even in patients with normal kidney function, are now being reported. A review of these data provides the foundation for a better understanding of kidney disease and, hopefully, better treatment for patients with HIV. PMID- 20005487 TI - The pathogenesis of HIV-associated nephropathy. AB - HIV-associated nephropathy (HIVAN) is one of the leading causes of ESRD in HIV-1 seropositive patients. Patients typically present with heavy proteinuria and chronic renal failure with pathologic findings of collapsing focal segmental glomerulosclerosis (FSGS). The disease is caused by direct infection of renal epithelial cells by HIV-1 in a genetically susceptible host. The genetic factors responsible for the susceptibility to HIVAN among blacks include a noncoding variant in the podocyte-expressed gene nonmuscle myosin, heavy chain 9 (MYH9) as well as other genes yet to be identified. Podocyte and tubular dysfunction results from the expression of viral genes, in particular nef and vpr, and the subsequent dysregulation of numerous host factors, including critical signaling pathways, inflammatory mediators, and others. The identification of these factors has the potential to provide novel therapeutic targets to prevent and treat this important disease. PMID- 20005486 TI - Screening for chronic kidney disease in HIV-infected patients. AB - With improved survival afforded by highly active antiretroviral therapy (HAART), CKD has emerged as one of the primary comorbid conditions affecting HIV-infected individuals. Although CKD in HIV-infected individuals is classically thought of as a consequence of advanced HIV infection such as in the case of HIV-associated nephropathy, several factors likely contribute to the development CKD in HIV infection. These factors include genetic predisposition, age-related decline in kidney function, HAART-related metabolic changes, exposure to multiple nephrotoxic medications, and concurrent conditions such as hepatitis C or illicit drug use. Similar to the general population, proteinuria and impaired kidney function are associated with faster progression to acquired immune deficiency syndrome and death. Given the prevalence and impact of kidney disease on the course of HIV infection and its management, current guidelines recommend screening all HIV-infected individuals for kidney disease. This review focuses on the current guidelines for kidney disease screening and discusses traditional as well as promising strategies for detecting CKD in this vulnerable population. PMID- 20005488 TI - Gene-gene and gene-environment interactions in HIV-associated nephropathy: A focus on the MYH9 nephropathy susceptibility gene. AB - HIV-associated nephropathy (HIVAN) is a leading cause of ESRD in African Americans. The HIV-1 virus infects podocytes, cells integral to formation of the glomerular filtration barrier, often leading to focal segmental glomerulosclerosis. HIVAN is typically a complication of late-stage HIV infection, associated with low CD4 cell counts and elevated serum HIV RNA levels. Highly active antiretroviral therapy is partially protective and has altered the natural history of HIV-associated kidney disease. Nonetheless, HIVAN remains an important public health concern among HIV-infected African Americans. Although polymorphisms in the MYH9 gene on chromosome 22 are strongly associated with HIVAN, as well as with idiopathic focal segmental glomerulosclerosis and global glomerulosclerosis (historically labeled "hypertensive nephrosclerosis"), the majority of HIV-infected patients who are genetically at risk from MYH9 do not appear to develop severe kidney disease. Therefore, we postulate that additional environmental exposures and/or inherited factors are necessary to initiate human HIVAN. Gene-environment interactions have also been proposed as necessary for the initiation of HIVAN in murine models. It is important that these novel risk factors be identified because prevention of environmental exposures and targeting of additional gene products may reduce the risk for HIVAN, even among those harboring 2 risk alleles in MYH9. PMID- 20005489 TI - Diagnosis and natural history of HIV-associated nephropathy. AB - HIV-associated nephropathy (HIVAN) is a largely distinctive phenotype induced by HIV-1 infection and is the most recognized and detrimental kidney disease in HIV infected patients. Host and viral characteristics have been implicated in the pathogenesis of HIVAN that may explain its exclusive predilection to patients of African descent. In untreated patients, the disorder is clinically manifested by an acute decline in kidney function, most often in conjunction with high-grade proteinuria and uncontrolled HIV-1 infection. Histologically, proliferating glomerular epithelial cells are the prominent feature of the disease. Data have evolved over the past decade suggesting that highly active antiretroviral therapy (HAART) can change the natural history of HIVAN, not only by preventing its development but also by halting its progression once developed. Consequently, with the widespread use of HAART, the prevalence of HIVAN is declining in Western countries. In contrast, the epidemiology of the disease is not well defined in the poorest areas in the world, which bear a disproportionate share of the HIV-1 epidemic's burden. Corticosteroids and inhibition of the renin-angiotensin axis are recommended as adjunctive agents in treating patients with established HIVAN and are potentially helpful in delaying the need for renal replacement therapy. However, the long-term value and potential risks of using corticosteroids in this population are unclear. PMID- 20005490 TI - The treatment of HIV-associated nephropathy. AB - Antiretroviral therapy (ART) preserves kidney function in patients with human immunodeficiency virus (HIV)-associated nephropathy (HIVAN). Emerging data also document substantial renal benefits of ART in the general HIV-infected population, which is associated in part with suppression of HIV-1 viral replication. The extent to which the response to ART differs in persons with HIVAN compared with those with other HIV-associated kidney disorders is unknown. Beneficial effects of corticosteroids and angiotensin-converting enzyme inhibitors on kidney function also are suggested by retrospective cohort studies and uncontrolled trials of patients with HIVAN. Underexposure to ART or inadequate ART dosing in HIV-infected patients with CKD may curtail the optimal benefits that may be derived from this therapy. PMID- 20005491 TI - Antiretroviral medications: adverse effects on the kidney. AB - The widespread introduction of highly active antiretroviral therapy (HAART) in the mid-1990s dramatically altered the course of human immunodeficiency virus (HIV) infection, with improvements in survival and reductions in the incidence of AIDS-defining illnesses. Although antiretroviral therapy has been shown to reduce the incidence of both AIDS-defining and non-AIDS conditions, long-term exposure to HAART may also be associated with significant toxicity. This article reviews the potential nephrotoxicity of specific antiretroviral agents and the impact of antiretroviral therapy on related metabolic disorders. The antiretroviral agents most strongly associated with direct nephrotoxicity include the nucleotide reverse transcriptase inhibitor, tenofovir, and the protease inhibitor indinavir, although other agents have been implicated less frequently. Tenofovir and related nucleotide analogs have primarily been associated with proximal tubular dysfunction and acute kidney injury, whereas indinavir is known to cause nephrolithiasis, obstructive nephropathy, and interstitial nephritis. Kidney damage related to antiretroviral therapy is typically reversible with early recognition and timely discontinuation of the offending agent, and nephrologists should be familiar with the potential toxicity of these agents to avoid delays in diagnosis. PMID- 20005492 TI - CKD in HIV-infected patients other than HIV-associated nephropathy. AB - A spectrum of kidney diseases in HIV-infected patients has been reported both before and after the introduction of highly active antiretroviral therapy (HAART). Kidney syndromes affecting HIV-infected patients include CKD as well as proteinuria, nephrotic syndrome, and acute nephritic syndrome. Thrombotic microangiopathy should be considered in patients with kidney disease and typical clinical characteristics. As the HIV-infected population ages, there is increased concern regarding the incidence of vascular and metabolic disease, leading to an increased burden of CKD. Although HIV-associated nephropathy is still the major cause of nephrotic syndrome in HIV-infected patients, immune complex glomerulonephritis (ICGN) still comprises a substantial proportion of the disease burden, especially in people of European origin. Genetic investigations into the underpinnings of the various histologic expressions of HIV-associated kidney disease hold great promise. The single most important diagnostic test to differentiate various forms of kidney disease in HIV-infected patients is a kidney biopsy. The results of treating kidney disease in HIV-infected patients remain unclear, and properly designed randomized controlled trials of the treatment of ICGN with HAART and other approaches are desperately needed. PMID- 20005493 TI - Kidney transplantation in patients with HIV infection. AB - HIV infection is a common cause of ESRD, particularly among blacks. Advances in antiretroviral therapy have greatly improved the survival of HIV patients, including those with renal disease. Despite concerns about the risk of immunosuppressive medications for HIV patients, emerging studies have now reported acceptable short-term outcomes for eligible HIV recipients undergoing renal transplantation, and an ongoing multicenter clinical trial reported 1-year patient and graft survival similar to that of HIV-uninfected kidney recipients. In these studies, the interactions between calcineurin inhibitors and HIV medications that are also metabolized by the cytochrome P450 system required substantial dosing modifications and careful monitoring of calcineurin inhibitor trough levels. These studies also revealed an elevated risk of acute rejection of the kidney allograft but few reports of opportunistic infections or viremia that could not be controlled. Long-term follow-up studies will be important to examine these outcomes and the development of malignancy to fully evaluate the risk and benefits of kidney transplantation among HIV-positive recipients. PMID- 20005494 TI - Management of HIV-infected patients with ESRD. AB - Patients infected with human immunodeficiency virus (HIV) often progress to ESRD. In the era of highly active antiretroviral therapy, the care of these patients has become increasingly complex as survival has improved. Patients infected with HIV who also have ESRD are at risk for critical interactions between medication regimens to treat both of these conditions. Within this population, hemo- and peritoneal dialysis as well as kidney transplantation are life sustaining but present a host of obstacles related to HIV monitoring and risk of transmission, access thrombosis, infection, and rejection. Knowledge of antiretroviral regimens, drug interactions, and HIV resistance as well as the management of ESRD in the presence of HIV infection will improve the care of these unique patients. PMID- 20005495 TI - A national survey of evolving management patterns for vascular injury. AB - BACKGROUND: The modern era has witnessed an increase in endovascular techniques used by physicians to treat vascular injury and age-related disease. As a consequence, the number of open vascular operations available for general surgical education has decreased dramatically. This changing paradigm threatens competence in vascular injury management achieved during surgical residency. The objective of this study is to sample perceptions on vascular injury treatment in the United States to highlight the need for planning for this important tenet of surgical education. METHODS: An electronic survey was extended to board-certified surgeons through 3 professional societies, the Peripheral Vascular Surgery Society (PVSS), the Eastern Association for the Surgery of Trauma (EAST), and the American College of Surgeons (ACS). RESULTS: A total of 520 respondents were self categorized as trauma (59%; n = 307), vascular (17%; n = 90), or general (19%; n = 99) surgeons. Respondents reported that general surgeons currently manage less than 10% of vascular injuries at their respective institutions. A 2.5-fold increase in endovascular treatment of vascular injury during the past decade was reported with interventional radiologists now involved in the management of up to 25% of injuries. Few general or trauma surgeons surveyed possessed a catheter based skill set, although 38% of trauma surgeons expressed great interest in endovascular training. Additionally, a cadre of vascular surgeons (67%) affirmed a commitment to teaching vascular injury management. CONCLUSIONS: The results of this study confirm a diminished role for non-fellowship-trained surgeons in managing vascular injury. Despite an increased acceptance of endovascular techniques to manage trauma, general and trauma surgeons do not possess the skill set. Collaboration between surgical communities will be especially important to maintain high standards in vascular injury management. PMID- 20005496 TI - A novel critical skills curriculum for surgical interns incorporating simulation training improves readiness for acute inpatient care. AB - INTRODUCTION: Surgical interns encounter complex, acute care situations often managed with limited supervision. Furthermore, medical school training does not adequately prepare students for special surgical considerations. Using simulation training, we implemented a course aimed at improving surgical intern readiness for responding to unique, life-threatening issues encountered in daily surgical care. METHODS: Twenty University of Minnesota surgical interns participated in the 3-week course. The first session consisted of interactive didactics and simulation covering hypoxia, shock, and metabolic disturbances; the second session addressed cardiopulmonary emergencies, including ventricular assist device and pacemaker use. Electronic simulation scenarios comprised the third session, allowing learners to demonstrate learned/practiced skills. The outcomes were assessed objectively (pretest and posttest) and subjectively (standardized feedback evaluations). RESULTS: Fifteen learners completed the pretest and posttest. The mean absolute score increase was 14% with average relative score improvement of 43%. Twenty learners completed feedback evaluations using a standard 5-point Likert scale. Respondents scored the first 2 sessions on topic importance (5 = very important), giving the first session 4.90 (+/- 0.31) and the second session 4.45 (+/- 0.89). Respondents ranked their confidence in executing practiced skills on actual patients (5 = very confident) as 4.24 (+/- 0.71). There was uniform support for the value of the electronic simulation scenarios as enhanced learning tools. CONCLUSIONS: We developed a course for surgical interns incorporating didactics and simulation. Learners demonstrated objective improvement in testing and reported that the course topics were highly important. After course completion, learners provided feedback indicating a high level of confidence in executing practiced skills, suggesting improved preparation for acute surgical care. PMID- 20005497 TI - Continuous, data-rich appraisal of surgical trainees' operative abilities: a novel approach for measuring performance and providing feedback. AB - BACKGROUND: We developed a convenient mechanism, Surgical Training and Assessment Tool (STAT), to accomplish detailed, continuous analysis of surgical trainees' operative abilities, and a simple method, Quality Based Surgical Training (QBST) for implementing it. METHODS: Using a web-accessed computer program, attending physicians and trainees independently assessed the trainee's operative performance after every operative (training) case. Global attributes of surgical knowledge, skill, and independence were assessed as well as the key technical maneuvers of each operation. A system of hierarchical, expandable menus specific to each of hundreds of different surgical procedures allowed the assessments to be made as detailed or as general as the users felt were necessary. In addition, freehand, unscripted commentary was recorded via an optional "remarks" box feature. Finally, an independently chosen, "overall" grade scaled F through A+ concluded each assessment. RESULTS: Over a 31 month period, 72 different users (52 trainees, 20 attending physicians) submitted 3849 performance assessments on 2424 cases, including 132 different case types and amassing 68,260 distinct data points. The mean number of data points per trainee was 1313; the median time spent per assessment was 60 seconds. Graphic displays allowed formative review of individual cases in real time, and summative review of long term trends. Appraisals of knowledge, skill, and independence were strongly correlated with and independently predictive of the overall competency grade (model r(2) = 0.68; test of predictive significance p < 0.001 for each rating). Trainee and attending physician scores were highly correlated (> 0.7) with one another. CONCLUSIONS: QBST/STAT achieves detailed, continuous analysis of surgical trainees' operative abilities, and facilitates timely, specific, and thorough feedback regarding their performance in theater. QBST/STAT promotes trainee self-reflection and generation of continuous, transparent, iterative training goals. PMID- 20005498 TI - Perirectal abscess infections related to MRSA: a prevalent and underrecognized pathogen. AB - BACKGROUND: Methicillin-resistant Staphylococcus aureus (MRSA) is not a commonly recognized pathogen isolated from perirectal abscesses. Complex perirectal abscesses of MRSA origin may present a significant challenge to the physician and result in treatment failure. The aim of our study was to determine the prevalence of MRSA in our patient population with perirectal abscesses and whether antibiotics coverage, if given, was adequate. METHODS: We conducted a retrospective study of all adult patients who presented with a perirectal abscess. The bacteriology of the cultures taken from more than 100 patients during an 8-year period was examined. The prevalence of MRSA was specifically analyzed, and the sensitivities of the organisms isolated were then compared with any antibiotics prescribed to determine the adequacy of coverage. RESULTS: In all, 124 patients were treated for perirectal abscess during the 8-year period. Fifty-nine percent of patients were taken to the operating room for incision and drainage, 52% of the patients had cultures taken of the abscess, and 35% of patients were given antibiotics coupled with incision and drainage. The antibiotics offered adequate coverage when compared with the sensitivities of the organisms 73% of the time. The prevalence of MRSA in our patient population was 19%, and when this organism was cultured, the patient only received adequate coverage 33% of the time. CONCLUSION: The presence of MRSA in perirectal abscesses is underrecognized. Recent data has shown that incision and drainage combined with antibiotics offers a superior outcome in soft tissue abscesses caused by this organism. If cultures are not routinely taken at the time of incision and drainage, the institutional incidents of MRSA will remain unknown. Physicians must recognize that MRSA is a potential organism present in perirectal abscesses when considering antimicrobial therapy for complex abscesses. PMID- 20005499 TI - Sentinel lymph node biopsy in breast cancer: a comprehensive literature review. AB - Sentinel lymph node biopsy has emerged as the new standard of care for nodal staging in early-stage breast disease. In the this review, the procedure of SLNB in breast cancer will be examined in greater detail with the aim of understanding techniques that may improve results and of identifying future research questions in this field. PMID- 20005500 TI - Suture end length as a function of knot integrity. AB - OBJECTIVE: To evaluate tension at the failure of 3 commonly used sutures when suture ends were cut to 3 lengths. STUDY DESIGN: Knots were tied using U.S. Pharmacopeia Size 0-0 polyglactin 910, silk, or polydioxanone sutures. The knots were tied randomly on a jig by the same surgeon. End lengths were then cut to random lengths of 0, 3, and 10 mm. We compared the individual knot strength when subjected to tensile forces via tensiometer with the point of knot failure, which was defined as untying and/or breaking of the knot. RESULTS: Three types of suture were divided into 3 groups based on end lengths for a total of 178 knots. A logistic regression analysis showed the odds of knots coming untied were highest for polyglactin 910 (odds ratio [OR] = 33.7; 95% confidence interval [CI] = 4.1-277.1). End length also had a significant effect on knots untying, with the 0-mm knots being more likely to come untied (OR, 21.2; 95% CI, 2.9-153.0). Post hoc tests for a 3 x 3 analysis of variance found that silk knots failed at significantly lower tension than polydioxanone (p < 0.001) and polyglactin 910 (p < 0.001) knots. CONCLUSIONS: The knots with an end length of 0 mm were significantly more likely to come untied than either 3- or 10-mm knots. Among all the materials, polyglactin 910 was the most prone to untying; however, it resulted in untying at a mean tension greater than the breaking point of silk. PMID- 20005501 TI - Surgical practice: evidence or anecdote. AB - OBJECTIVES: Our objective is to highlight a few surgical practices that are not based on evidence but are still taught in surgical education, and to assess our experience with these practices. DESIGN: We identified 3 practices (clamping of nasogastric tubes before removal, bowel preparation before elective colon resection, and elective sigmoid colectomy following 2 bouts of diverticulitis), identified the data supporting each practice, and administered a survey to faculty and residents at our institution. SETTING: Wright State University Department of Surgery, Boonshoft School of Medicine, Dayton, Ohio. PARTICIPANTS: Twenty-one faculty and 35 residents responded to the survey. RESULTS: No studies were found relating to clamping nasogastric tubes before removal. Seven faculty (33%) and 11 residents (31%) used this practice. Two faculty (10%) and 0 residents felt this was an evidence-based practice. Faculty were more likely to have reviewed the evidence (85% vs 40%, p < 0.001). Level 2 evidence has shown bowel preparation did not improve outcomes relating to anastomotic leak, wound infection, or septic complications in elective colon resection. Twenty faculty (95%) and 34 residents (97%) used this practice. Faculty were more likely to believe this to be evidence-based (85% vs 49%, p = 0.01). There has been no level 1 or 2 evidence showing that sigmoid colectomy following 2 bouts of diverticulitis improves morbidity or mortality. Fourteen faculty (70%) and 26 residents (76%) reported using this practice. Twelve faculty (60%) and 21 residents (60%) felt this was evidence-based. CONCLUSIONS: Frequent use of surgical practices without evidence support can create a misperception that such practices are evidence-based. Faculty are more likely to change a practice after obtaining continuing medical education, suggesting that residents may need validation by faculty practice of evidence-based procedures before incorporation into their clinical care. PMID- 20005502 TI - Tarp's baker's dozen: instructions to beginning program directors and some lessons learned. PMID- 20005503 TI - The effect of a focused instructional session on knowledge of surgical staplers in general surgery residents. AB - INTRODUCTION: Surgical stapling devices have been used for a variety of purposes in both laparoscopic and open surgery. Nevertheless, trainees rarely receive any focused instruction on their application and use. This study attempts to determine the baseline knowledge of surgical stapling devices possessed by surgical residents. Furthermore, we attempt to evaluate the effectiveness of a short didactic session in improving the trainee's knowledge of the use and function of surgical staplers. METHODS: A 20-question multiple-choice test was created to evaluate a general surgery resident's knowledge on the design and use of circular, linear, and laparoscopic surgical staplers. The test was administered before and after attending a 40-minute instructional session on surgical stapling devices. The tests were then scored by a data analyst. RESULTS: A total of 26 residents of 39 in the residency program (26/39, 67%) participated. The pretest mean was 10.62/20 (53%), whereas the posttest mean was 15.38/20 (77%). These results were significantly different on paired samples t-test analysis (t((25)) = -10.3; p < 0.05). The mean pretest scores were also significantly different between resident levels (R1-R2, 9.50; R3-R5, 11.31; t((24)) = -2.10; p < 0.05). Senior-level residents scored higher on posttest analysis, but this result was not significant (R1-R2, 14.70; R3-R5, 15.81; t((24)) = -1.63; p > 0.05). DISCUSSION: There is a deficiency of knowledge of surgical staplers in general surgery residents, more so in junior residents. Didactic instruction is effective in raising the level of knowledge of surgical staplers in all residents, up to a similar level. Surgical educators should consider implementing programs like these for staplers and other types of surgical equipment. PMID- 20005504 TI - Scholarly activity. PMID- 20005505 TI - Retained wireless video enteroscopy capsule: a case report and review of the literature. AB - Capsule enteroscopy, which is a wireless noninvasive approach to evaluation of the small intestine, consists of an 11 x 26-mm capsule containing a miniature video camera, batteries, illuminating light-emitting diodes, a transmitter, and an antenna. The components inside the capsule are encased by a slippery, nonbiodegradable, plastic housing, which weighs less than 4 g. Information obtained from the imager, approximately 2 frames per second, is transmitted by way of radiotelemetry to an array of sensors taped to the abdomen, which connect to a data recorder worn on a belt around the waist. The batteries are designed to last roughly 7-8 hours, providing enough energy to generate approximately 50,000 to 60,000 detailed images. The clinical review of the imagery is made available after completion of the study when images are downloaded from the data recorder to a computer with dedicated software. PMID- 20005506 TI - Two-surgeon couple. PMID- 20005507 TI - Maternal and neonatal morbidities associated with obstructive sleep apnea complicating pregnancy. AB - OBJECTIVE: The objective of the study was to estimate the maternal and neonatal morbidities associated with obstructive sleep apnea (OSA) in pregnancy. STUDY DESIGN: Women delivering between 2000-2008 with confirmed OSA in an academic center were included. Normal-weight and obese controls were randomly selected at a 2:1 ratio. Maternal and neonatal morbidities were compared between the groups. Multivariate analyses were performed to evaluate maternal morbidity and preterm birth (PTB). RESULTS: The analysis included 57 pregnancies complicated by OSA. Compared with normal-weight (n = 114) controls, OSA patients had more preeclampsia (PET) (19.3% vs 7.0%; P = .02) and PTB (29.8% vs 12.3%; P = .007). Controlling for comorbid conditions, OSA was associated with an increased risk of PTB (odds ratio [OR], 2.6; 95% confidence interval [CI], 1.02-6.6), mostly secondary to PET (63%). Cesarean delivery (OR, 8.1; 95% CI, 2.9-22.1) and OSA were associated with maternal morbidity (OR, 4.6; 95% CI, 1.5-13.7). CONCLUSION: Pregnancies complicated by OSA are at increased risk for preeclampsia, medical complications, and indicated PTB. PMID- 20005508 TI - Management of acute Stevens-Johnson syndrome and toxic epidermal necrolysis utilizing amniotic membrane and topical corticosteroids. AB - PURPOSE: To describe the results of a novel treatment approach to the acute ophthalmic management of Stevens-Johnson syndrome (SJS) and toxic epidermal necrolysis (TEN). DESIGN: Retrospective interventional case series. METHODS: setting: Institutional. study population: Sixteen eyes of 8 patients with acute, biopsy-proven SJS or TEN and significant ophthalmic involvement. interventional procedure(s): Application of amniotic membrane to the ocular surface, either in the operating room or at the bedside, and short-term use of intensive topical corticosteroid medication. main outcome measures: Visual acuity, slit-lamp appearance of the ocular surface, and patients' subjective impression of ocular comfort. RESULTS: Two patients expired during the hospitalization. Mean follow-up time for the surviving patients was 7.7 months. Four surviving patients in whom the entire ocular surface (ie, the cornea, bulbar and palpebral conjunctiva, and eyelid margins) was treated with amniotic membrane retained visual acuities of 20/40 or better and an intact ocular surface. In contrast, the initial 2 patients in the study who were treated with only a Prokera device or unsutured amniotic membrane sheets, leaving the palpebral conjunctiva and eyelid margins uncovered, developed more significant ocular surface abnormalities, and 1 developed a corneal perforation. CONCLUSIONS: Amniotic membrane coverage of the ocular surface in its entirety coupled with the use of intensive short-term topical corticosteroids during the acute phase of SJS and TEN is associated with the preservation of good visual acuity and an intact ocular surface. Partial amniotic membrane coverage of the ocular surface may not serve to minimize the cicatrizing ocular sequelae of SJS and TEN as effectively as complete coverage. PMID- 20005509 TI - Vitamin C attenuates the cytotoxic effects of Porphyromonas gingivalis on human gingival fibroblasts. AB - OBJECTIVE: Periodontitis is induced by an imbalance between bacterial virulence and host defense ability involving increased levels of oxidative stress. The aim of this study was to investigate the influence of vitamin C on the cytotoxic effects of Porphyromonas gingivalis on human gingival fibroblasts (HGF). METHODS: This in vitro study observed the interaction between HGF and P. gingivalis. HGF were cultured with medium containing vitamin C and exposed to P. gingivalis ATCC 33277 for a maximum of 180 min. The assessment of cell viability was followed by a 3-(4,5-dimethylthiazol-2-ly)-2,5-diphenyltetrazolium-bromide (MTT) assay. The apoptosis rate was detected by flow cytometry using Annexin-V-FITC and propidium iodide. Superoxide as an oxidative stress factor was measured photometrically by the reduction of ferricytochrome C. RESULTS: Vitamin C reduced the cytotoxic effects of P. gingivalis on HGF. Vitamin C-treated HGF showed significantly higher cell viability rates (89.0+/-5.7%) in comparison to untreated HGF (77.0+/ 5.0%; p<0.05). In vitamin C-treated HGF, lower apoptosis rates (40.0+/-2.2%) were observed after P. gingivalis exposure than in untreated HGF (66.1+/-1.6%; p<0.05). The exposure of HGF to P. gingivalis led to a significant increase of superoxide concentration, but this effect was not influenced by vitamin C. CONCLUSION: Vitamin C reduces the cytotoxic and apoptotic effects of P. gingivalis on HGF in vitro. These results suggest that the benefit of vitamin C should be further investigated clinically. PMID- 20005510 TI - Molecular characterization of Streptococcus mutans strains containing the cnm gene encoding a collagen-binding adhesin. AB - OBJECTIVE: Streptococcus mutans, known to be a major pathogen of dental caries, is also considered to cause infective endocarditis. Its 120-kDa Cnm protein binds to type I collagen, which may be a potential virulence factor. In this study, we characterized S. mutans clinical strains focusing on the cnm gene encoding Cnm. DESIGN: A total of 528 S. mutans strains isolated from Japanese, Finnish, and Thai subjects were investigated. Using molecular techniques, the distribution frequency of cnm-positive strains and location of the inserted cnm were analyzed. Furthermore, isogenic mutant strains were constructed by inactivation of the cnm gene, then their biological properties of collagen-binding and glucan-binding were evaluated. Southern hybridization of the genes encoding glucan-binding proteins was also performed. RESULTS: The distribution frequency of cnm-positive strains from Thai subjects was 12%, similar to that previously reported for Japanese and Finnish subjects. Furthermore, the location of insertion of cnm was the same in all cnm-positive clinical isolates. As for the cnm-inactivated mutant strains constructed from 28 clinical isolates, their collagen-binding activity was negligible. In addition, glucan-binding activity in the cnm-positive clinical isolates was significantly reduced and corresponded to a lack of gbpA encoding glucan-binding protein A. CONCLUSIONS: Our results indicate that strains with cnm genes, the most crucial factor for the collagen-binding property of S. mutans, are detectable at similar frequencies over several different geographic locations. In addition, the common properties of these strains are a high level of collagen-binding activity and tendency for a low level of glucan-binding activity. PMID- 20005511 TI - Matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry analysis of oligosaccharides and oligosaccharide alditols obtained by hydrolysis of agaroses and carrageenans, two important types of red seaweed polysaccharides. AB - MALDI-TOF mass spectrometry analyses of several oligosaccharides (aldoses) and oligosaccharide alditols derived from agaroses, kappa- and iota-carrageenans using different matrices (2,5-dihydroxybenzoic acid, nor-harmane, ferulic acid, and the ionic liquid matrices 2,5-dihydroxybenzoic acid-n-butylamine and ferulic acid-n-butylamine) were conducted. These carbohydrates were selected as model compounds to study the MALDI prompt and post-source decay (PSD) fragmentation processes of both families of oligosaccharides. Sulfated alditols showed in the negative-ion mode the molecular ion as [M-Na](-) together with the species yielded by their prompt fragmentation (mainly desulfation) while the sulfated oligosaccharides (aldoses) showed mainly glycosidic prompt fragmentation (glycosidic C-cleavages and desulfation). Non-sulfated aldoses and alditols, which could only be analyzed in positive-ion mode ([M+Na](+)), did not suffer any prompt fragmentation. The former yielded cross-ring fragmentation in the PSD mode. Best results were obtained by using 2,5-dihydroxybenzoic acid and/or nor harmane as matrices for all the compounds studied. PMID- 20005512 TI - Identifiability of fractional order systems using input output frequency contents. AB - In this paper, issues related to the identifiability of a fractional order system having its input and output frequency contents are discussed. The effects of the commensurate order alpha in the identifiability of the model structure and model parameters are analytically studied. It is shown that both identifiabilities (model structure and model parameters) are reduced remarkably for smaller values of alpha. This phenomenon is observed even though the input signals are rich enough and system belongs to the model set. Our understanding is that the problem arises since differences among different members of the model set fall beyond the practically recognizable precision range. The issue is more problematic when alpha is smaller and measurements are noisy. PMID- 20005513 TI - Health-related quality of life and life satisfaction in patients following surgically treated pelvic ring fractures. A prospective observational study with two years follow-up. AB - BACKGROUND: Pelvic ring fractures caused by high-energy trauma are severe injuries with well described radiological and clinical outcomes, whereas description from the patient's perspective is less well documented. The purpose of this study was to investigate patient-reported outcome following surgical treatment of pelvic fractures using quality of life instruments. METHODS: All 54 patients (28 male/26 female, ages 16-68) with pelvic fractures referred to our institution for surgical treatment 2003-2005 were prospectively included. The most common trauma was motor vehicle accident (44%). Additional injuries were seen in 74% and in 31% the ISS was >or=16. There were 31 B and 23 C type fractures. Patients were followed for two years using two validated questionnaires, SF-36 and LiSat-11, the latter an instrument consisting of 11 questions for evaluation of satisfaction with different aspects of life. RESULTS: 45 patients could be followed according to the study protocol for two years while 2 were untraceable and 1 died from unrelated causes. Of 6 nonresponders, 5 were unable due to psychiatric disorder. At two years pelvic fracture patients scored lower than the reference population in both physical and mental domains (SF-36). Highest mean score, 68, was in the domain Social Function (norm 89) while lowest mean score, 38, was in the domain Role Physical (norm 86). The mean score closest to the normative was for general health with 61 for patients and 78 for the normative group. In LiSat-11 pelvic fracture patients scored lower than the reference population in all areas. Satisfaction with life as a whole was 31% compared with 60% in the normative group. CONCLUSIONS: Two years after surgical treatment of pelvic ring fractures, patients reported substantially lower quality of life for both physical and mental domains, when compared with a reference population, even when radiological and clinical outcomes were considered favourable. PMID- 20005514 TI - Postchallenge plasma glucose excursions, carotid intima-media thickness, and risk factors for atherosclerosis in Chinese population with type 2 diabetes. AB - AIMS: Isolated hyperglycemia is associated with atherosclerosis in individuals with type 2 diabetes, but the relationship between postchallenge glucose excursion and atherosclerosis is less clear. This study examines the relationships between postchallenge glucose spikes (PGS), carotid intima-media thickness (IMT), and traditional risk factors for atherosclerosis in individuals with type 2 diabetes. METHODS: A total of 474 individuals with type 2 diabetes who were within the highest or lowest IMT distribution quartile were included. The Student's t-test, one-way analysis of variance (ANOVA), single variate and multivariate analyses were implemented to study the data. An additional healthy control group (n=896) was selected during routine health examination. They were Han nationality and unrelated to the diabetic patients. RESULTS: (1) Compared with subjects of healthy control group, the subjects with type 2 diabetes had significantly higher levels of body mass index (BMI), waist-to-hip ratio (WHR), systolic blood pressure, triglyceride, total cholesterol, low density lipoprotein cholesterol (LDL-C), fasting plasma glucose (FPG), 120 min postchallenge glucose (PG120), hemoglobin A1c (HbA1c) and IMT (P < or = 0.01) and relatively lower levels of high density lipoprotein cholesterol (HDL-C) (P < or = 0.05). (2) According to the IMT which was measured by B-mode ultrasonography, the patients of type 2 diabetes could be divided into two subgroups: one was the subgroup of IMT > or = P(75) and another was the subgroup of IMT < or = P(25). Compared with subjects of IMT > or = P(25) subgroup, subjects being in the IMT > or = P(75) subgroup exhibited significantly increased age, WHR, diabetes duration, systolic blood pressure, total cholesterol, triglyceride, LDL-C, and significantly decreased HDL-C levels. And among all the plasma glucose variables, except for FPG and PG30, all the other variables (include PG60, PG120, PG180, PGS, HbA1C, under area curve of glucose) showed a significant increase in the IMT>/=P(75) subgroup. (3) A multivariate logistic regression analysis was performed to establish which were independently related with carotid IMT, and the results showed the PGS was identified as the strongest determinant of IMT from all the atherosclerosis risk factors. (4) PGS is significantly correlated to a variety of atherosclerosis risk factors. CONCLUSIONS: This study identified several important associations between PGS and known risk factors for atherosclerosis and suggested that PGS is independently related to carotid IMT. Wide postchallenge glucose excursions may contribute to the development of atherosclerosis in individuals with type 2 diabetes, independent of other risk factors. PMID- 20005515 TI - The effect of PPAR-alpha agonism on apolipoprotein metabolism in humans. AB - Metabolic syndrome, diabetes and obesity are frequently associated with hypertriglyceridemia, hypercholesterolemia and low HDL levels, a phenotype known as atherogenic dyslipidemia. Atherogenic dyslipidemia and hypertriglyceridemia are frequently treated with fibric acid derivatives which activate the nuclear receptor PPAR-alpha leading to reduce plasma triglycerides and an increase in HDL cholesterol levels. The mechanism by which activation of PPAR-alpha with fibrates improves the plasma lipid profile in patients with atherogenic dyslipidemia and hypertriglyceridemia has been examined in several small studies measuring lipoprotein kinetics. The results of these studies indicate that the changes in lipoprotein metabolism observed in response to fibrate treatment vary according to lipoprotein phenotype. In general, fibrates act to reduce VLDL apoB-100 through enhanced fractional catabolism (clearance) of VLDL apoB-100 with additional effects on reducing VLDL apoB-100 production. LDL apoB-100 levels generally decrease in response to fibrates due to increased LDL fractional catabolism except in those patients with high to very high plasma triglyceride levels (>400mg/dL). Fibrates also increase HDL apoA-I and apoA-II levels by enhancing apoA-I and apoA-II production, although this is partially counteracted by increasing fractional catabolism of these apolipoproteins. The potent and specific PPAR-alpha agonist LY518674, reduced VLDL apoB-100 levels through enhanced fractional catabolism similar to what is seen with fibrates. In contrast to fibrates, LY518674 did not change HDL apoA-I levels in response to due to an increased turnover of apoA-I where an increased fractional catabolic rate entirely counteracted the increase in apoA-I production. The changes in apoB metabolism in response to PPAR-alpha activation with fibrates and specific PPAR alpha agonists would be expected to reduce the risk of cardiovascular disease. However, the benefit of the enhanced turnover of HDL apoA-I in response to PPAR alpha activation remains to be determined. PMID- 20005516 TI - Lipoprotein-associated phospholipase A(2), inflammatory biomarkers, and risk of cardiovascular disease in the Prospective Study of Pravastatin in the Elderly at Risk (PROSPER). AB - OBJECTIVE: Lipoprotein-associated phospholipase A(2) (Lp-PLA(2)) is an inflammatory biomarker that circulates mainly bound to LDL. We evaluated the association of Lp-PLA(2) with vascular events in the elderly where the importance of LDL is diminished as a risk factor for coronary disease. METHODS: Mass and activity of Lp-PLA(2) were related to risk over 3.2 years for vascular events (definite or suspected death from CHD, non-fatal MI, fatal or non-fatal stroke) in the 2804 men and 3000 women age 70-82 years in the Prospective Study of Pravastatin in the Elderly (PROSPER). RESULTS: Lp-PLA(2) showed a moderate, positive association with risk of a vascular event with hazard ratios of 1.25 (confidence interval (CI) 1.02-1.54) for mass and 1.39 (CI 1.14-1.70) for activity for top versus bottom quartile. Risk associations were attenuated when classical risk factors, lipids and inflammatory markers - C-reactive protein and white cell count - were included in the models. Lp-PLA(2) was unrelated to stroke risk. Inclusion of all three inflammatory markers in multivariate models negated the association of HDL cholesterol with risk (hazard ratio 0.98; CI 0.88-1.10) and increased prediction of coronary events; the C statistic rose from 63.2% to 64.4% (P<0.001). CONCLUSION: In elderly people Lp-PLA(2), alongside other inflammatory indices, is a potential biomarker for vascular events, particularly CHD. PMID- 20005517 TI - Using digital image correlation to determine bone surface strains during loading and after adaptation of the mouse tibia. AB - Previous models of cortical bone adaptation, in which loading is imposed on the bone, have estimated the strains in the tissue using strain gauges, analytical beam theory, or finite element analysis. We used digital image correlation (DIC), tracing a speckle pattern on the surface of the bone during loading, to determine surface strains in a murine tibia during compressive loading through the knee joint. We examined whether these surface strains in the mouse tibia are modified following two weeks of load-induced adaptation by comparison with contralateral controls. Results indicated non-uniform strain patterns with isolated areas of high strain (0.5%), particularly on the medial side. Strain measurements were reproducible (standard deviation of the error 0.03%), similar between specimens, and in agreement with strain gauge measurements (between 0.1 and 0.2% strain). After structural adaptation, strains were more uniform across the tibial surface, particularly on the medial side where peak strains were reduced from 0.5% to 0.3%. Because DIC determines local strains over the entire surface, it will provide a better understanding of how strain stimulus influences the bone response during adaptation. PMID- 20005518 TI - Functional properties of cartilaginous tissues engineered from infrapatellar fat pad-derived mesenchymal stem cells. AB - Articular cartilage has a poor intrinsic capacity for self-repair. The advent of autologous chondrocyte implantation has provided a feasible method to treat cartilage defects. However, the associated drawbacks with the isolation and expansion of chondrocytes from autologous tissue has prompted research into alternative cell sources such as mesenchymal stem cells (MSCs) which have been found to exist in the bone marrow as well as other joint tissues such as the infrapatellar fat pad (IFP), synovium and within the synovial fluid itself. In this work we assessed the chondrogenic potential of IFP-derived porcine cells over a 6 week period in agarose hydrogel culture in terms of mechanical properties, biochemical content and histology. It was found that IFP cells underwent robust chondrogenesis as assessed by glycosaminoglycan (1.47+/-0.22% w/w) and collagen (1.44+/-0.22% w/w) accumulation after 42 days of culture. The 1Hz dynamic modulus of the engineered tissue at this time point was 272.8 kPa (+/ 46.8). The removal of TGF-beta3 from culture after 21 days was shown to have a significant effect on both the mechanical properties and biochemical content of IFP constructs after 42 days, with minimal increases occurring from day 21 to day 42 without continued supplementation of TGF-beta3. These findings further strengthen the case that the IFP may be a promising cell source for putative cartilage repair strategies. PMID- 20005519 TI - An innovative application of a small-scale motion analysis technique to quantify human skin deformation in vivo. AB - This study highlights a new experimental method developed to measure full-field deformation of human skin in vivo. The technique uses a small-scale Qualisys (Sweden) 3D motion capture system and an array of reflective markers placed on the forearm of five healthy volunteers. A load of up to 1.5N was applied to induce skin deformation by pulling a fine wire attached to the centre of the marker configuration. Loading and marker displacements were recorded simultaneously. 3D marker trajectory data was generated for three different load directions. Tests were repeated to investigate accuracy and repeatability. Calibration results indicate the accuracy of the motion capture system with an average residual of 0.05 mm. The procedure was found to be repeatable and accurate for five repeated tests of measured displacements with a maximum variance of 5%. Experimental data are presented to demonstrate robustness and the ability to produce significant outputs. For all five subjects, at 1N load, the mean and standard deviations of skin axial and lateral displacements were found to be 11.7+/-1.6mm and 12.3+/-3.3mm, respectively. The axial displacements ratio (u(90)/u(0)) ranges from 0.63 to 1.45 with mean+/-standard deviation of 0.982+/ 0.34 and 0.982+/-0.32 for left and right arms, respectively. The experiments generated useful and accurate data that can be used to study the viscoelastic, hyperelastic or anisotropic behaviour of human skin. The measured displacements will be analysed further to determine the mechanical properties of skin using inverse Finite Element Analysis and Ogden model. PMID- 20005520 TI - Application of ultrasound-assisted extraction to the determination of contaminants in food and soil samples. AB - The application of ultrasound-assisted extraction (UAE) to the sample preparation of environmental and food samples has increased in the last years. This technique has been used in the development of methods for the analysis of numerous contaminants, including organic compounds (pesticides, pharmaceuticals, polycyclic aromatic hydrocarbons, polyhalogenated flame retardants, etc.) and heavy metals. The aim of this work is to review the application of this extraction procedure to the analysis of contaminants in food and soil and the comparison of its use with other well-established extraction procedures. The advantages and disadvantages of this technique together with the possibility of coupling UAE with other analytical techniques will be also discussed. PMID- 20005521 TI - Evolution of dispersive liquid-liquid microextraction method. AB - Dispersive liquid-liquid microextraction (DLLME) has become a very popular environmentally benign sample-preparation technique, because it is fast, inexpensive, easy to operate with a high enrichment factor and consumes low volume of organic solvent. DLLME is a modified solvent extraction method in which acceptor-to-donor phase ratio is greatly reduced compared with other methods. In this review, in order to encourage further development of DLLME, its combination with different analytical techniques such as gas chromatography (GC), high performance liquid chromatography (HPLC), inductively coupled plasma-optical emission spectrometry (ICP-OES) and electrothermal atomic absorption spectrometry (ET AAS) will be discussed. Also, its applications in conjunction with different extraction techniques such as solid-phase extraction (SPE), solidification of floating organic drop (SFO) and supercritical fluid extraction (SFE) are summarized. This review focuses on the extra steps in sample preparation for application of DLLME in different matrixes such as food, biological fluids and solid samples. Further, the recent developments in DLLME are presented. DLLME does have some limitations, which will also be discussed in detail. Finally, an outlook on the future of the technique will be given. PMID- 20005522 TI - Stability analysis of an encapsulated microbubble against gas diffusion. AB - Linear stability analysis is performed for a mathematical model of diffusion of gases from an encapsulated microbubble. It is an Epstein-Plesset model modified to account for encapsulation elasticity and finite gas permeability. Although bubbles, containing gases other than air, are considered, the final stable bubble, if any, contains only air, and stability is achieved only when the surrounding medium is saturated or oversaturated with air. In absence of encapsulation elasticity, only a neutral stability is achieved for zero surface tension, the other solution being unstable. For an elastic encapsulation, different equilibrium solutions are obtained depending on the saturation level and whether the surface tension is smaller or higher than the elasticity. For an elastic encapsulation, elasticity can stabilize the bubble. However, imposing a non-negativity condition on the effective surface tension (consisting of reference surface tension and the elastic stress) leads to an equilibrium radius which is only neutrally stable. If the encapsulation can support a net compressive stress, it achieves actual stability. The linear stability results are consistent with our recent numerical findings. Physical mechanisms for the stability or instability of various equilibriums are provided. PMID- 20005523 TI - Hydrothermal synthesis and luminescent properties of YVO4:Ln(3+) (Ln = Eu, Dy, and Sm) microspheres. AB - Rare-earth ions (Eu(3+), Dy(3+), Sm(3+)) doped YVO(4) microspheres with uniform morphologies were successfully prepared via a simple hydrothermal route using N,N dimethylformamide (DMF) as the solvent and polyvinylpyrrolidone (PVP) as protective agent. X-ray diffraction (XRD), scanning electron microscopy (SEM), transmission electron microscopy (TEM), photoluminescence (PL) spectra, and the kinetic decays were employed to examine the resulting phase formation, particle morphology and luminescent properties. The XRD results reveal that all the doped samples are of high crystallization which are assigned to the pure tetragonal phase of YVO(4). Additionally, the DMF/H(2)O volume ratio and the concentration of PVP both have obvious effects on the morphologies and sizes of the as synthesized products. The sample prepared at 180 degrees C for 24 h with the DMF/H(2)O volume ratio of 3/1 and 0.4 g/L PVP concentration exhibits uniformly spherical shape with the diameter of 1-2 microm. Upon excitation by ultraviolet radiation or low-voltage electron beams excitation, the YVO(4):Ln(3+) (Ln=Eu, Dy, and Sm) samples show strong light emissions with different colors from the doped Ln(3+) ions. These phosphors exhibit potential applications in the fields of fluorescent lamps and light emitting diodes (LEDs). PMID- 20005524 TI - Effects of audio-visual information on the intelligibility of alaryngeal speech. AB - BACKGROUND: There is minimal research on the contribution of visual information on speech intelligibility for individuals with a laryngectomy (IWL). AIMS: The purpose of this project was to determine the effects of mode of presentation (audio-only, audio-visual) on alaryngeal speech intelligibility. METHOD: Twenty three naive listeners were randomly presented 176 phonemically balanced sentences produced by three alaryngeal (tracheoesophageal, esophageal, electrolaryngeal) and one typical, laryngeal speaker in both audio-only and audio-visual modes of presentation. RESULTS: Overall, results showed a small (3%) but statistically significant higher speech intelligibility score for audio-visual stimuli compared to audio-only stimuli as well as a significant interaction effect between mode of speech and mode of presentation. Within mode of speech, electrolaryngeal speech was the only mode that benefited significantly from the inclusion of visual information (10% increase). Overall intelligibility showed similar patterns both within audio-only and audio-visual modes of presentation: typical laryngeal and tracheoesophageal speech modes were significantly more intelligible than both esophageal and electrolaryngeal. CONCLUSIONS: Results suggest that listeners may benefit more from visual information from speakers with poor baseline speech intelligibility. Results also show similar intelligibility between typical, laryngeal and tracheoesophageal modes of speech. Results should be interpreted with caution, however, as only one speaker from each mode of speech was included. Further research is required to determine the nature of the increase. LEARNING OUTCOMES: Readers will (1) increase their understanding of the potential impact of visual information in the perception of alaryngeal speech; (2) identify potential factors that may augment or hinder speech perception; and (3) discuss how individual modes of alaryngeal speech may be affected by audio-visual information. PMID- 20005525 TI - The Second Merial European Comparative Vaccinology Symposium. PMID- 20005526 TI - Ageing, immunosenescence and inflammageing in the dog and cat. AB - Improvements in veterinary healthcare over recent decades mean that we now have a significant population of geriatric small companion animals. The design of optimum nutritional and vaccination programmes for these aged animals must be underpinned by knowledge of the physiological changes that occur in later life. It is clear that older dogs and cats are affected by the process of immunosenescence and that similar changes occur in these species to those documented in elderly people. The most consistent findings of recent investigations indicate impairment of cell-mediated immune function with age. Senior dogs and cats are generally shown to have reduced blood CD4(+) T cells (with imbalance in Th1 versus Th2 functional activity), elevation in the CD8(+) subset and reduction in the CD4:CD8 ratio. The ability of blood lymphocytes to respond to stimulation by mitogens decreases, as does the cutaneous delayed type hypersensitivity response. By contrast, there is relative preservation of the ability to mount humoral immune responses. Serum and salivary immunoglobulin (Ig)A production increases and IgG concentration remains unaltered with age. Elderly animals generally have persisting vaccinal antibody titres at protective level and respond to booster vaccination with elevation in titre. Older dogs and cats are able to make primary humoral responses to novel antigens, but the magnitude of these may be reduced relative to titres achieved in younger animals. Fewer investigations have studied the phenomenon of 'inflammageing' (the effect of cumulative antigenic exposure and onset of late life inflammatory disease) in these species. Senior cats have increased production of pro-inflammatory cytokines by blood monocytes, but this effect has not been demonstrated with cells derived from older dogs. Numerous studies have investigated whether canine and feline immunosenescence might be slowed or reversed by dietary supplementation with antioxidants, but no significant research has addressed the need for geriatric vaccination protocols. PMID- 20005527 TI - What's in the name? Or how rocks and stones are different from bunnies and rabbits. AB - Labels have been shown to play an important role in inductive generalization; however, the mechanism by which labels contribute to generalization early in development remains unclear. We investigated two factors that may influence the inductive potential of labels: semantic similarity and co-occurrence probability. Results suggested that adults and 6-year-olds rely on semantic similarity of labels and that their generalizations are not affected by co-occurrence probability. Specifically, generalization patterns were qualitatively similar for co-occurring semantically similar labels (e.g., bunny-rabbit) and non-co occurring semantically similar labels (e.g., rock-stone) in 6-year-olds and adults. Unlike 6-year-olds and adults, 4-year-olds were likely to generalize co occurring labels but not non-co-occurring labels. Possible mechanisms by which co occurrence probability may influence label generalization in young children are discussed. PMID- 20005528 TI - Nonelective cardiac surgery in the elderly: is it justified? AB - OBJECTIVE: Elderly patients might be denied nonelective cardiac surgery because of the perception of poor outcomes and an unacceptable quality of life. In this study we evaluate long-term survival and quality of life in these patients. METHODS: From 1994 to 1999, 262 consecutive patients older than 80 years underwent urgent (n = 223) or emergent (n = 39) cardiac surgery. Of these patients, 160 (61%) underwent coronary artery bypass grafting, 64 (24%) underwent coronary artery bypass grafting plus valve surgery, 17 (7%) underwent valve surgery, and 21 (8%) underwent aortic surgery. Kaplan-Meier survival analysis and quality-of-life assessment were performed, and result were compared with age adjusted population data. Risk factors for mortality were determined by using Cox regression. The utility of Society of Thoracic Surgeons and EuroSCORE risk scoring were assessed by using area under receiver operating curves. RESULTS: Early mortality was 11% (n = 29) overall, 7% (n = 16) in urgent cases, and 33% (n = 13) in emergent cases. Five-year survival was 50% (n = 132) overall, 53% (n = 105) in urgent cases, and 36% (n = 18) in emergent cases. There was no difference in 10-year survival between patients undergoing urgent surgical intervention and age-adjusted population data. Among survivors, quality-of-life measures were equivalent to those of the general elderly population. Risk factors for early mortality were age, emergent procedure, aortic procedure, bypass time, and postoperative complication (renal failure, myocardial infarction, cerebrovascular accident, pneumonia, and reoperation for bleeding). Risk factors for late mortality were peripheral vascular disease, emergent procedure, bypass time, and new renal failure. The EuroSCORE and Society of Thoracic Surgeons risk scores were equivalent but only moderately predictive of mortality. CONCLUSIONS: Long term survival and quality of life after nonelective cardiac surgery can equal that of the general elderly population. Age alone should not disqualify a patient for urgent or emergent cardiac surgery. PMID- 20005529 TI - A follow-up of integrated positron emission tomography/computed tomography after curative resection of non-small-cell lung cancer in asymptomatic patients. AB - OBJECTIVE: A follow-up integrated positron emission tomography/computed tomography (PET/CT), as part of a more intensive surveillance program, has been performed at around 1 year after curative resection, regardless of the patients' symptoms or findings in other tests. This study was designed to evaluate the results of this follow-up integrated PET/CT in patients treated for non-small cell lung cancer without symptoms or abnormal findings. METHODS: Between January 2003 and December 2006, this study enrolled 86 patients with non-small cell lung cancer who had no clinical or radiologic evidence of recurrence after curative resection before integrated PET/CT and underwent a follow-up integrated PET/CT around 1 year at our institution. RESULTS: The time from operation to the follow up integrated PET/CT check was 13.4 +/- 4.4 months. Integrated PET/CT showed negative findings in 41 (47.7%) patients, equivocal findings in 16 (18.6%) patients, and positive findings in 29 (33.7%) patients. Twenty-seven (31.4%) patients had recurrent disease and 2 patients had extrathoracic double primary cancer. Six patients had extrathoracic recurrence without intrathoracic recurrence. CONCLUSIONS: A postoperative follow-up integrated PET/CT can be used for early detection of recurrence in asymptomatic patients who had had resection of non-small-cell lung cancer. Further studies are required to evaluate the cost effectiveness or survival benefit of follow-up integrated PET/CT. PMID- 20005530 TI - Midterm outcomes of myocardial revascularization in children. AB - OBJECTIVE: Pediatric coronary artery bypass grafting is uncommon. Small target vessels and appropriate conduit choice are the main technical challenges. METHODS: Fourteen patients undergoing coronary artery bypass grafting from January 1986 to December 2008 were retrospectively reviewed. RESULTS: Median age was 10 years (range, 3-15 years); median weight was 36 kg (range, 12-71 kg). Indications included symptoms or evidence of inducible ischemia and angiographically documented coronary stenosis. Diagnoses included Kawasaki disease (5/14), anomalous left coronary artery originating from the pulmonary artery (2/14), previous stent implant (1/14), and metabolic disease (3/14). The remaining 3 patients had coronary stenosis after other cardiac operations. Preoperatively 5 patients (45%) had no symptoms and 9 (64%) had positive stress test. Single-vessel disease was demonstrated in 2 (14%), double-vessel disease in 7 (50%), triple-vessel disease in 1 (7%), and left main coronary artery involvement in 4 (29%). With standard cardiopulmonary bypass, 18 (81%) in situ internal thoracic arteries and 4 (19%) long saphenous veins were grafted. There was 1 early reoperation for graft failure. All patients survived to hospital discharge. Follow-up angiography was performed in 5 patients (36%; median, 2 years; range, 1 day-10 years), and 1 (7%) required late balloon dilatation. Median follow-up was 3.3 years (1 month-10 years), and 12 patients had no symptoms. There was 1 late death of noncardiac cause. CONCLUSIONS: Pediatric coronary artery bypass grafting can be performed for a wide range of indications. Midterm results are excellent. Preoperative stress testing can detect silent myocardial ischemia. PMID- 20005531 TI - Exploring the marine biotic index (AMBI): variations on a theme by Angel Borja. AB - The results of manipulating the input data to AMBI are explored using various transformations of numerical species abundance (NAMBI), biomass (BAMBI) and production (PAMBI) from a variety of stations on the NE Atlantic shelf at which the pollution/disturbance status is known. There is a close agreement between the proportions of species in the five AMBI ecological groups and a phylum level meta analysis axis of increasing environmental impact. All AMBI measures provide a better monotonic relationship with the impact axis than do traditional species diversity measures, which show higher diversity at intermediate levels of disturbance. A marginally better relationship with the impact axis of the meta analysis is achieved by a moderate (square root) transformation of the data. 'Production' data (an appropriate combination of abundance and biomass information) provide only a marginal improvement on abundance data, but are ecologically and functionally much more relevant. Severe transformation of the data, culminating in presence/absence, degrades the relationship with the impact axis, but if only simple species lists are available then these may still be useable in making an environmental assessment. PMID- 20005532 TI - Identification of petroleum hydrocarbons using a reduced number of PAHs selected by Procrustes rotation. AB - Identifying petroleum-related products released into the environment is a complex and difficult task. To achieve this, polycyclic aromatic hydrocarbons (PAHs) are of outstanding importance nowadays. Despite traditional quantitative fingerprinting uses straightforward univariate statistical analyses to differentiate among oils and to assess their sources, a multivariate strategy based on Procrustes rotation (PR) was applied in this paper. The aim of PR is to select a reduced subset of PAHs still capable of performing a satisfactory identification of petroleum-related hydrocarbons. PR selected two subsets of three (C(2)-naphthalene, C(2)-dibenzothiophene and C(2)-phenanthrene) and five (C(1)-decahidronaphthalene, naphthalene, C(2)-phenanthrene, C(3)-phenanthrene and C(2)-fluoranthene) PAHs for each of the two datasets studied here. The classification abilities of each subset of PAHs were tested using principal components analysis, hierarchical cluster analysis and Kohonen neural networks and it was demonstrated that they unraveled the same patterns as the overall set of PAHs. PMID- 20005533 TI - The Gulf: a young sea in decline. AB - This review examines the substantial changes that have taken place in marine habitats and resources of the Gulf over the past decade. The habitats are especially interesting because of the naturally high levels of temperature and salinity stress they experience, which is important in a changing world climate. However, the extent of all natural habitats is changing and their condition deteriorating because of the rapid development of the region and, in some cases from severe, episodic warming episodes. Major impacts come from numerous industrial, infrastructure-based, and residential and tourism development activities, which together combine, synergistically in some cases, to cause the observed deterioration in most benthic habitats. Substantial sea bottom dredging for material and its deposition in shallow water to extend land or to form a basis for huge developments, directly removes large areas of shallow, productive habitat, though in some cases the most important effect is the accompanying sedimentation or changes to water flows and conditions. The large scale of the activities compared to the relatively shallow and small size of the water body is a particularly important issue. Important from the perspective of controlling damaging effects is the limited cross-border collaboration and even intra-country collaboration among government agencies and large projects. Along with the accumulative nature of impacts that occur, even where each project receives environmental assessment or attention, each is treated more or less alone, rarely in combination. However, their combination in such a small, biologically interacting sea exacerbates the overall deterioration. Very few similar areas exist which face such a high concentration of disturbance, and the prognosis for the Gulf continuing to provide abundant natural resources is poor. PMID- 20005534 TI - Prevalence of thyroid dysfunctions in systemic lupus erythematosus. AB - The association of systemic lupus erythematosus (SLE) and thyroid autoimmunity has been reported by several studies in a wide range of variability. However, from a review of the literature, discrepant results have been reported. The aim of the study was to evaluate the prevalence of clinical and subclinical thyroid disorders in patients with SLE vs sex- and age-matched controls. Thyroid hormones and the presence of antithyroid antibodies were tested and thyroid ultrasonography was performed in 213 patients with SLE vs 426 sex- and age matched controls, from the same geographic area, with a well-defined status of iodine intake. The odds ratio for subclinical hypothyroidism for female patients with SLE with respect to controls was 4.5 (95% confidence interval [CI], 2.5 8.4); for antithyroid peroxidase antibody (AbTPO) positivity, it was 2.6 (95% CI, 1.7-4.1); and for thyroid autoimmunity, it was 2.9 (95% CI, 2.0-4.4). The mean values of thyroid-stimulating hormone and AbTPO were higher in female SLE patients than in controls (P < .01). A significantly (P < .01) higher prevalence of clinical hypothyroidism and Graves disease was observed in female SLE patients than in controls. No significant difference between SLE patients and controls was detected with regard to free triiodothyronine and thyroxine. In our series, 3% of SLE patients had "nonthyroidal illness syndrome" vs 0 control. Thyroid function and AbTPOs should be tested and ultrasonography should be performed as part of the clinical profile in SLE patients. Subjects at high risk (women, positive AbTPOs, hypoechoic, and small thyroid) should have thyroid function follow-up and appropriate treatment in due course. PMID- 20005535 TI - The first and second phase of insulin secretion in naive Chinese type 2 diabetes mellitus. AB - Impaired insulin secretion (ISEC) has been recognized as one of the most important pathophysiologies of type 2 diabetes mellitus. There are 2 phases of ISEC: the first phase (first ISEC) and second phase (second ISEC). This study aimed to evaluate the 2 phases of ISEC in newly diagnosed type 2 diabetes mellitus patients. Fifty-two drug-naive type 2 diabetes mellitus patients were given 2 tests: a modified low-dose graded glucose infusion (M-LDGGI) and frequent sample intravenous glucose tolerance test. The M-LDGGI is a simplified version of the Polonsky method. Two stages of intravenous infusion of glucose with different rates were given, starting from 2 mg/(kg min) and then followed by 6 mg/(kg min). Each stage was maintained for 80 minutes. The results were interpreted as the slope of the changes of plasma insulin against the glucose levels. The slope of these curves was regarded as the second ISEC and used as the criterion for grouping-the responders and nonresponders. The responders are older and had higher body mass index and log (homeostasis model assessment of beta-cell function) (log HOMA-beta) but lower fasting plasma glucose and hemoglobin A(1c) (HbA(1c)) than the nonresponders. Significant correlations were only noted between the second ISEC and first ISEC (r = 0.278, P = .046) and between the second ISEC and log HOMA-beta (r = 0.533, P = .000). Correlation between different parameters and HbA(1c) was also evaluated. Only second ISEC and log HOMA-beta were correlated significantly with HbA(1c) (r = -0.388, P = .015 and r = -0.357, P = .026, respectively). In type 2 diabetes mellitus, subjects with higher second ISEC are older and have higher body mass index. At the same time, second ISEC is the most important factor for determining glucose levels in naive Chinese type 2 diabetes mellitus patients. The first and second ISECs were only modestly correlated, which indicated that the deterioration of these 2 phases was not synchronized. Finally, we also recommend using the M-LDGGI for quantifying second ISEC. This practical method could be done in many centers without difficulty. PMID- 20005536 TI - Serum uric acid and metabolic syndrome in Taiwanese adults. AB - A positive association between serum uric acid and metabolic syndrome has been reported, but little information is available about the association between serum uric acid and metabolic syndrome in Taiwanese adults. The purpose of this study was to investigate the association between serum uric acid levels and metabolic syndrome in Taiwanese adults. We performed a cross-sectional study of 2085 men and 1557 women. All of the participants underwent a health screening during the period from January 2005 to December 2005 at a health center of the Shin Kong Wu Ho-Su Memorial Hospital. Metabolic syndrome was defined according to the National Cholesterol Education Program Adult Treatment Panel III criteria. The results showed that hyperuricemia was significantly associated with increased risk for hypertriglyceridemia, low high-density lipoprotein cholesterol level, and high blood pressure in men and women. The risk of metabolic syndrome was significantly higher in the fourth quartile than in the first quartile of uric acid level in men (odds ratio [OR], 1.50; 95% confidence interval [CI], 1.06-2.14) and women (OR, 2.33; 95% CI, 1.39-3.93). In addition, uric acid level was inversely associated with hyperglycemia in men. The ORs of hyperglycemia for the second, third, and fourth quartile of uric acid were 0.69 (95% CI, 0.46-1.03), 0.55 (95% CI, 0.37-0.83), and 0.45 (95% CI, 0.29-0.69), respectively, compared with the lowest quartile of uric acid. The results demonstrate that there is a positive association between serum uric acid levels and metabolic syndrome and an inverse association between uric acid and fasting plasma glucose in Taiwanese adults. PMID- 20005538 TI - Is the interaction between HIF1A P582S and ACTN3 R577X determinant for power/sprint performance? AB - Hypoxia-inducible factor 1 (HIF-1) is a transcription factor that regulates gene expression in response to hypoxia and has been associated with athletic performance. The aims of this study were (1) to determine the frequency distribution of HIF1A Pro582Ser (rs11549465) polymorphism among 155 Israeli athletes (sprinters and endurance athletes) and 240 healthy controls and (2) to analyze the influence of the interaction between HIF1A Pro582Ser and ACTN3 R577X (rs1815739) genotypes on sprint performance. There were no differences across the HIF1A genotype and allele frequencies among endurance athletes, sprinters, and controls. Similarly, no differences were found between the subgroups of top-level and national-level endurance athletes, or between top-level and national-level sprinters. Conversely, interaction effects were found between HIF1A Pro582Ser and ACTN3 R577X polymorphisms and sprinters. The proportion of HIF1A Pro/Pro + ACTN3 R/R genotypes was significantly higher in sprinters than in endurance athletes and healthy controls (P = .002). In addition, the odds ratio for HIF1A Pro/Pro + ACTN3 R/R genotype carriers being a sprinter was 2.25 (95% confidence interval, 1.24-4.1); and that for HIF1A Pro/Pro + ACTN3 R/R genotype carriers being an endurance athlete was 0.5 (95% confidence interval, 0.2-1.24). We conclude that HIF1A Pro582Ser polymorphism by itself is not critical in determining sprint performance. However, sprinter performance is determined by the interaction between the wild-type HIF1A Pro/Pro genotype and ACTN3 RR genotype. PMID- 20005539 TI - Clinical observation paper: fatty liver and metabolic syndrome: is it a burden for the future generations? PMID- 20005540 TI - Increased leptin storage with altered leptin secretion from adipocytes of rats with sucrose-induced dyslipidemia and insulin resistance: effect of dietary fish oil. AB - This study examined the effect of long-term feeding a high-sucrose diet (SRD) on the modulation of rat adipocyte's leptin secretion and storage. For this purpose, we analyzed (a) basal and insulin-stimulated leptin release and the role of isoproterenol and palmitate on insulin-stimulated leptin secretion, (b) the correlation between leptin and glycerol released, (c) the relationship between leptin contents and adiposity, and (d) the effect of fish oil (FO) administration on the above parameters. Wistar rats were fed an SRD for 6 months. Whereas half the animals continued with SRD up to month 8, the other half was fed an SRD in which FO partially replaced corn oil from months 6 to 8. Total leptin release was reduced both basally and under insulin stimulation in SRD-fed rats. However, the ratio of leptin released after hormone stimulation to basal leptin levels was similar in the 3 dietary groups. Isoproterenol inhibited insulin-stimulated leptin release in the 3 groups, but the percentage was lower in the SRD. Palmitic acid mimicked the effect of isoproterenol. Leptin release from adipocyte of SRD fed rats negatively correlated with glycerol release. Leptin store increased in fat pads of SRD and positively correlated with adiposity. Fish oil reduced leptin content and fat pad hypertrophy, and normalized basal lipolysis, leptinemia, and glucose homeostasis. This suggests that enhanced lipolysis and altered insulin sensitivity could play a role in the decrease of leptin released in SRD-fed rats. This is consistent with the reversion of all the alterations after FO administration. PMID- 20005541 TI - CYP7A1 (-204 A>C; rs3808607 and -469 T>C; rs3824260) promoter polymorphisms and risk of gallbladder cancer in North Indian population. AB - Cholesterol 7-alpha hydroxylase (CYP7A1), which is a rate-limiting enzyme for cholesterol catabolism and bile acid synthesis, may affect cholesterol homeostasis and result in gallstone formation that is a major risk factor for gallbladder cancer (GBC) pathogenesis. Genetic variations in CYP7A1 may influence its expression and thus may affect the risk of gallstone disease and GBC. We aimed to study the association of 2 promoter polymorphisms of CYP7A1 (-204 A>C [rs3808607] and -469 T>C [rs3824260]) in gallstone and GBC susceptibility in North Indian population. The study included 185 GBC patients, 195 symptomatic gallstone patients, and 200 healthy controls. Genotyping for both polymorphisms was done by polymerase chain reaction-restriction fragment length polymorphism method. Although the CC genotype of CYP7A1 -204 A>C was not significantly associated with gallstone disease (P = .083, odds ratio [OR] = 1.69, 95% confidence interval [CI] = 0.9-3.0), it was conferring higher risk for GBC (P = .018, OR = 2.05, 95% CI = 1.1-3.7). However, CYP7A1 -469 T>C was not associated with gallstone disease and GBC risk in our population. After subgroup stratifications on the basis of sex and gallstone status, CC genotype and variant allele of CYP7A1 -204 A>C imparted higher risk for GBC in women (P = .003, OR = 3.30, 95% CI = 1.5-7.2) and patients without gallstones (P = .045, OR = 1.91, 95% CI = 1.2-3.6). Haplotype analysis of the 2 polymorphisms showed that C,T (P = .045, OR = 1.84, 95% CI = 1.0-3.3) and C,C (P = .0001, OR = 3.10, 95% CI = 1.6 6.0) haplotypes had elevated risk of GBC predisposition. CYP7A1 -469 T>C is not associated with gallstone disease or GBC risk. Although CYP7A1 -204 A>C might play a modest role in gallstone susceptibility, it is an independent risk factor for GBC in North Indian population. Underlying mechanism for GBC susceptibility by CYP7A1 (-204 A>C and -469 T>C) haplotype appears to be independent of gallstone pathway and is believed to involve genotoxicity resulting from subnormal bile acid production. PMID- 20005542 TI - Fasting cholesteryl ester transfer protein concentration is independently associated with the postprandial decrease in high-density lipoprotein cholesterol concentration after fat-rich meals: the Hoorn prandial study. AB - The aim of the study was to test whether fasting or postprandial cholesteryl ester transfer protein (CETP) concentrations are associated with postprandial changes in high-density lipoprotein cholesterol (HDL-c) concentrations after fat rich or carbohydrate-rich meals. Postmenopausal women (76 with normal glucose metabolism [NGM], 41 with type 2 diabetes mellitus [T2DM], and 38 T2DM women with statin therapy [T2DM-ST]) received 2 consecutive fat-rich or carbohydrate-rich meals on separate occasions. Linear regression analysis was performed to assess the associations of fasting CETP and postprandial changes of CETP with postprandial changes in HDL-c. Mean plasma HDL-c concentrations decreased significantly after the fat-rich meals: 0.18 +/- 0.09 mmol/L in NGM, 0.16 +/- 0.09 mmol/L in T2DM, and 0.14 +/- 0.08 mmol/L in T2DM-ST women. This effect was smaller after using carbohydrate-rich meals: 0.12 +/- 0.09 mmol/L in the NGM, 0.12 +/- 0.08 mmol/L in the T2DM, and 0.10 +/- 0.05 mmol/L in the T2DM-ST study group. Higher fasting but not postprandial CETP concentrations were associated with a larger postprandial decrease in HDL-c (beta -0.034; 95% confidence interval, -0.067 to -0.001) after the fat-rich meals. This association was independent of the postprandial increase in triglycerides and similar among the 3 study groups. A high fasting CETP concentration may contribute to the postprandial atherogenic lipoprotein profile in postmenopausal women by decreasing HDL-c after fat-rich meals. This effect is independent from the postprandial increase in triglycerides. PMID- 20005543 TI - Effect of nutrition on plasma C-type natriuretic peptide forms in adult sheep: evidence for enhanced C-type natriuretic peptide degradation during caloric restriction. AB - Previous studies in lambs and children show that the plasma concentration of amino terminal pro-C-type natriuretic peptide (NTproCNP), a stable product of proCNP, is strongly correlated with skeletal growth and markers of bone formation. Consistent with these findings, CNP expression is sensitive to nutritional status and is reduced by caloric restriction (CR) in both the fetus and the postnatal lamb. However, the effect of nutritional status on CNP in the adult, once linear growth is complete, is unknown. Hypothesizing that reduced CNP synthesis during CR is contingent on the presence of active growth plates, we studied the effect of CR ( 25% of maintenance) or loading (CL, 200% of maintenance) on CNP forms and alkaline phosphatase (ALP) in adult ewes and compared the findings to responses in a control group (C) fed a maintenance diet of 10.6 MJ of metabolizable energy. Live body weight was reduced (17%) in the CR group and increased (10%) in the CL group after 16 days of intervention. Plasma CNP concentration and ALP both fell in CR sheep and were significantly lower than C (P < .05 for both), returning toward basal levels 1 week after refeeding. In contrast, plasma NTproCNP did not differ (CR vs C). There were no significant changes in CNP forms and ALP in CL sheep compared with C. Fall in plasma CNP but not in NTproCNP in CR adult sheep suggests that CNP degradation (not synthesis) is altered, and contrasts with previous findings in growing lambs where CR reduces both CNP forms. PMID- 20005545 TI - Combination of two oxidant stressors suppresses the oxidative stress and enhances the heat shock protein 27 response in healthy humans. AB - We tested the hypothesis that the combination of 2 oxidant stressors (hyperoxia and fatiguing exercise) might reduce or suppress the oxidative stress. We concomitantly measured the plasma concentration of heat shock proteins (Hsp) that protect the cells against the deleterious effects of reactive oxygen species. Healthy humans breathed pure oxygen under normobaric condition for 50-minute periods during which they stayed at rest or executed maximal static handgrip sustained until exhaustion. They also repeated handgrip bouts in normoxic condition. We performed venous blood measurements of 2 markers of the oxidative stress (thiobarbituric acid reactive substances and reduced ascorbic acid) and Hsp27. Under normoxic condition, the handgrip elicited an oxidative stress and a modest increase in plasma Hsp27 level (+7.1 +/- 5.4 ng/mL). Under hyperoxic condition, (1) at rest, compared with the same time schedule in normoxic condition, we measured an oxidative stress (increased thiobarbituric acid reactive substances and decreased reduced ascorbic acid levels) and the plasma Hsp27 level increased (maximal variation, +12.5 +/- 6.0 ng/mL); and (2) after the handgrip, the oxidative stress rapidly disappeared. The combination of both hyperoxia and handgrip bout doubled the Hsp27 response (maximal variation, +24.8 +/- 9.2 ng/mL). Thus, the combination of 2 hits eliciting an oxidative stress seems to induce an adaptive Hsp27 response that might counterbalance an excessive production of reactive oxygen species. PMID- 20005544 TI - Effects of glucose and insulin on acyl ghrelin and desacyl ghrelin, leptin, and adiponectin in pregnant women with diabetes. AB - The aim of the study was to compare the regulation of ghrelin, leptin, and adiponectin by insulin and glucose during the second and third trimesters of pregnancy in women with diabetes. We studied 9 pregnant women with diabetes. All women were treated with insulin and omitted the morning dose on the day of the test. After collection of baseline fasting samples, we performed 3 successive glucose clamps: 2 euglycemic clamps (glucose, 5 mmol/L; insulin infusion at 20 and 40 mU m(-2) min(-1)) and 1 hyperglycemic clamp (glucose, 10 mmol/L; insulin infusion at 40 mU m(-2) min(-1)). We determined concentrations of acyl and desacyl ghrelin (using a double-antibody sandwich assay that recognizes the full length molecule), leptin, and adiponectin. Fasting desacyl ghrelin concentrations decreased, whereas insulin and leptin concentrations increased, between the second and third trimesters of pregnancy (P < or = .011). During the clamp studies, desacyl ghrelin concentrations decreased by 33% (second trimester, P = .004) and 27% (third trimester, P = .09) with increasing glucose and insulin concentrations, whereas acyl ghrelin, leptin, and adiponectin concentrations were unaffected. Glucose and insulin regulate desacyl ghrelin concentrations in pregnant women with diabetes. Impaired desacyl ghrelin regulation may affect energy metabolism in pregnant women with poorly controlled diabetes. PMID- 20005546 TI - Metabolic profiling of heat or anoxic stress in mouse C2C12 myotubes using multinuclear magnetic resonance spectroscopy. AB - In the present study, the metabolic effects of heat and anoxic stress in myotubes from the mouse cell line C2C12 were investigated by using a combination of (13)C, (1)H, and (31)P nuclear magnetic resonance (NMR) spectroscopy and enrichment with [(13)C]-glucose. Both the (13)C and the (1)H NMR spectra showed reduced levels of the amino acids alanine, glutamate, and aspartate after heat or anoxic stress. The decreases were smallest at 42 degrees C, larger at 45 degrees C, and most pronounced after anoxic conditions. In addition, in both the (1)H and the (31)P NMR spectra, decreases in the high-energy phosphate compounds adenosine triphosphate and phosphocreatine with increasing severity of stress were identified. At anoxic conditions, an increase in (13)C-labeled lactate and appearance of glycerol-3-phosphate were observed. Accumulation of lactate and glycerol-3-phosphate is in agreement with a shift to anaerobic metabolism due to inhibition of the aerobic pathway in the mitochondria. Conversely, lower levels of unlabeled ((12)C) lactate were apparent at increasing severity of stress, which indicate that lactate is released from the myotubes to the medium. In conclusion, the metabolites identified in the present study may be useful markers for identifying severity of stress in muscles. PMID- 20005547 TI - Proteomic approaches to study plant-pathogen interactions. AB - The analysis of plant proteomes has drastically expanded in the last few years. Mass spectrometry technology, stains, software and progress in bioinformatics have made identification of proteins relatively easy. The assignment of proteins to particular organelles and the development of better algorithms to predict sub cellular localization are examples of how proteomic studies are contributing to plant biology. Protein phosphorylation and degradation are also known to occur during plant defense signaling cascades. Despite the great potential to give contributions to the study of plant-pathogen interactions, only recently has the proteomic approach begun to be applied to this field. Biological variation and complexity in a situation involving two organisms in intimate contact are intrinsic challenges in this area, however, for proteomics studies yet, there is no substitute for in planta studies with pathogens, and ways to address these problems are discussed. Protein identification depends not only on mass spectrometry, but also on the existence of complete genome sequence databases for comparison. Although the number of completely sequenced genomes is constantly growing, only four plants have their genomes completely sequenced. Additionally, there are already a number of pathosystems where both partners in the interaction have genomes fully sequenced and where functional genomics tools are available. It is thus to be expected that great progress in understanding the biology of these pathosystems will be made over the next few years. Cheaper sequencing technologies should make protein identification in non-model species easier and the bottleneck in proteomic research should shift from unambiguous protein identification to determination of protein function. PMID- 20005548 TI - Sources of information for the use of periconceptual folic acid. PMID- 20005549 TI - [Cannabis-induced cerebral and myocardial infarction in a young woman]. AB - INTRODUCTION: Cannabis is the most consumed drug in the world particularly in young adults. Few reports have suggested a causal role of cannabis in the development of cerebral or cardiovascular events. We describe the first association of myocardial infarction and stroke after heavy cannabis consumption in a 45-year-old woman. OBSERVATION: Stroke occurred in relation with a right carotid and middle cerebral artery thrombosis after cannabis abuse. The patient was successfully treated with intravenous rt-PA. Two days after her admission, she presented a myocardial infarction due to a coronary thrombosis. Cerebral and coronary arteries were angiographically normal. Etiological tests were negative and a toxic cause in relation with cannabis consumption was concluded. CONCLUSION: Cannabis can be associated with vascular events by different mechanisms. Thrombosis may occur in cerebral and/or coronary arteries. We suggest that it might be useful to search for cannabis consumption systematically in young subjects victims of stroke and myocardial infarction. PMID- 20005550 TI - [Semantic knowledge in MCI and Alzheimer's disease: the French version of the New Words Interview]. AB - INTRODUCTION: The first decline in cognitive performance in Alzheimer's disease can appear when assessing semantic memory and can be detected long before the typical symptoms of Alzheimer's disease (AD), appearing with Mild Cognitive Impairment (MCI). PATIENTS AND METHOD: We propose the French version of the New Words Interview (fNWI) using 22 words to investigate semantic knowledge. The fNWI uses 11 words, which entered the French dictionary between 1996 and 1997, and 11 other words, which entered between 2006 and 2007. Words were paired according to orthographic and semantic criteria. Each word was associated with three sub tests: free evocation, discrimination of the best definition from three propositions, and recognition of the accurate word context (two sentences were proposed). Regarding evocation, we distinguished conceptual definition, life situation examples or examples by use. We tested 12 patients with AD, 12 patients with amnesic Mild Cognitive Impairment (aMCI) and 72 controls (12 were paired with patients for age and education level). RESULTS: MCI patients and AD patients exhibited lower performance than controls in the three sub-tests and for the words of both periods. From the early stage of MCI, the patients were more impaired in the fNWI than in the context recognition task, and they failed to provide conceptual definitions of new words. Therefore, MCI patients suffer from semantic impairments before obvious clinical signs of AD. CONCLUSION: In patients with AD, performance worsened on all subtests, and more strongly in the definition discrimination task, which suggests the impairment of stored semantic knowledge. They provided fewer conceptual definitions and failed to use the strategy observed in MCI patients, who compensated for conceptual difficulties by providing examples. PMID- 20005551 TI - Transverse oscillations for tissue motion estimation. AB - This paper gives an overview of the methods developed for tissue motion estimation using transverse oscillation images (TO). TO images are specific radiofrequency ultrasound images featuring oscillations in both spatial directions. The initial studies on TO were published in the late 1990s. This paper reviews the main ideas and applications behind this motion estimation approach. First the origin and motivation of TO is briefly reviewed. Then the beamforming methods that lead to TO images are given, detailing the receive-only approach and the transmit-and-receive approach using synthetic aperture data. The different medical applications where TO has been used are discussed (blood flow, elastography and echocardiography), showing how it can improve motion estimation. Finally, the future perspectives of TO are outlined. PMID- 20005552 TI - Clinical polyomavirus BK variants with agnogene deletion are non-functional but rescued by trans-complementation. AB - High-level replication of polyomavirus BK (BKV) in kidney transplant recipients is associated with the emergence of BKV variants with rearranged (rr) non-coding control region (NCCR) increasing viral early gene expression and cytopathology. Cloning and sequencing revealed the presence of a BKV quasispecies which included non-functional variants when assayed in a recombinant virus assay. Here we report that the rr-NCCR of BKV variants RH-3 and RH-12, both bearing a NCCR deletion including the 5' end of the agnoprotein coding sequence, mediated early and late viral reporter gene expression in kidney cells. However, in a recombinant virus they failed to produce infectious progeny despite large T-antigen and VP1 expression and the formation of nuclear virus-like particles. Infectious progeny was generated when the agnogene was reconstructed in cis or agnoprotein provided in trans from a co-existing BKV rr-NCCR variant. We conclude that complementation can rescue non-functional BKV variants in vitro and possibly in vivo. PMID- 20005554 TI - Bio-diatomite dynamic membrane reactor for micro-polluted surface water treatment. AB - This work investigated the feasibility of treating micro-polluted surface water for drinking water production with a bio-diatomite dynamic membrane reactor (BDDMR) at lab-scale in continuous-flow mode. Results indicate that the BDDMR was effective in removing COD(Mn), DOC, UV(254), NH(3)-N and trihalomethanes' formation potential (THMFP) at a hydraulic retention time (HRT) of 3.5h due to its high concentrations of mixed liquor suspended solids (MLSS) and mixed liquor volatile suspended solids (MLVSS). The removal of pollutants was mainly ascribed to microbial degradation in BDDMR because the dynamic membrane alone was much less effective in pollutant removal. Though the diatomite particles (5-20microm) were much smaller in size than the aperture of the stainless steel support mesh (74microm), microorganisms and their extracellular polymer substances could bind these particles tightly to form bio-diatomite particles which were completely retained by the support mesh. The analysis of molecular weight (MW) distribution by gel permeation chromatography (GPC) shows that the BDDMR could effectively remove the hydrophilic fraction of dissolved organic materials present in the raw water. PMID- 20005553 TI - Endocytic Rab proteins are required for hepatitis C virus replication complex formation. AB - During infection, hepatitis C virus (HCV) NS4B protein remodels host membranes to form HCV replication complexes (RC) which appear as foci under fluorescence microscopy (FM). To understand the role of Rab proteins in forming NS4B foci, cells expressing the HCV replicon were examined biochemically and via FM. First, we show that an isolated NS4B-bound subcellular fraction is competent for HCV RNA synthesis. Further, this fraction is differentially enriched in Rab1, 2, 5, 6 and 7. However, when examined via FM, NS4B foci appear to be selectively associated with Rab5 and Rab7 proteins. Additionally, dominant negative (DN) Rab6 expression impairs Rab5 recruitment into NS4B foci. Further, silencing of Rab5 or Rab7 resulted in a significant decrease in HCV genome replication. Finally, expression of DN Rab5 or Rab7 led to a reticular NS4B subcellular distribution, suggesting that endocytic proteins Rab5 and Rab7, but not Rab11, may facilitate NS4B foci formation. PMID- 20005555 TI - ESBL-producing E. coli in Austrian sewage sludge. AB - The aim of this study was to investigate the degree of contamination of sewage sludge with ESBL-producing Escherichia coli strains and the effectiveness of different sewage sludge treatment methods. Monthly sewage sludge samples were collected between January and September 2009 in 5 different sewage treatment plants and tested for the presence of ESBL E. coli. In addition, the number of colony forming units (CFU) of E. coli and coliform bacteria before and after the different sludge treatment methods (aerobic/anaerobic digestion, lime stabilization, and thermal treatment) was investigated. Of the 72 sewage sludge samples investigated, ESBL-positive E. coli were found in 44 (61.1%) sewage sludge samples. The classification of beta-lactamase groups was carried out in 15 strains resulting in the detection of 2 different groups (CTX-M and TEM) of bla genes. All 15 of them had a CTX-M gene and 4 of these strains furthermore carried a TEM gene. With regard to the CFU of E. coli and coliform bacteria, thermal treatment and lime stabilization following dehydration sufficiently reduced pathogen concentrations. The plants using merely stabilization and dehydration showed an increase of E. coli and coliform bacteria and thus also an increase in ESBL-producing E. coli. PMID- 20005556 TI - The effects of sub-lethal UV-C irradiation on growth and cell integrity of cyanobacteria and green algae. AB - The effects of UV-C irradiation on algal growth and cell integrity were investigated to develop a potential method for preventing cyanobacterial blooms. The toxic cyanobacterium Microcystis aeruginosa and three common freshwater green algae Chlorella ellipsoidea, Chlorella vulgaris, and Scenedesmus quadricanda were exposed to UV-C irradiation at 0-200mJcm(-2) and subsequently incubated for 9-15 d under normal culture conditions. Cell density and cell integrity were assessed using flow cytometry. The results suggested that UV-C irradiation at 20-200mJcm( 2) can suppress M. aeruginosa growth for 3-13 d in a dose-dependent manner. UV-C irradiation at 20 and 50mJcm(-2) is sub-lethal to M. aeruginosa cells as over 80% of the exposed cells remained intact. However, UV-C irradiation at 100 and 200mJcm(-2) induced severe cell disintegration in more than 70% of the irradiated cells. Neither significant suppression nor disintegration effects on green algae were observed for UV-C irradiation at 20-200mJcm(-2) in this study. Taken together, the sensitivity of M. aeruginosa to UV-C irradiation was significantly higher than that of the non-toxic C. ellipsoidea, C. vulgaris, and S. quadricauda, suggesting the potential application of sub-lethal UV-C irradiation for M. aeruginosa bloom control with a predictable low ecological risk. PMID- 20005557 TI - Eco-SpaCE: an object-oriented, spatially explicit model to assess the risk of multiple environmental stressors on terrestrial vertebrate populations. AB - Wildlife organisms are exposed to a combination of chemical, biological and physical stressors. Information about the relative impact of each stressor can support management decisions, e.g., by the allocation of resources to counteract those stressors that cause most harm. The present paper introduces Eco-SpaCE; a novel receptor-oriented cumulative exposure model for wildlife species that includes relevant ecological processes such as spatial habitat variation, food web relations, predation, and life history. A case study is presented in which the predicted mortality due to cadmium contamination is compared with the predicted mortality due to flooding, starvation, and predation for three small mammal species (Wood mouse, Common vole, and European mole) and a predator (Little owl) living in a lowland floodplain along the river Rhine in The Netherlands. Results indicated that cadmium is the principal stressor for European mole and Little owl populations. Wood mouse and Common vole population densities were mainly influenced by flooding and food availability. Their estimated population sizes were consistent with numbers reported in literature. Predictions for cadmium accumulation and flooding stress were in agreement with field data. The large uncertainty around cadmium toxicity for wildlife leads to the conclusion that more species-specific ecotoxicological data is required for more realistic risk assessments. The predictions for starvation were subject to the limited quantitative information on biomass obtainable as food for vertebrates. It is concluded that the modelling approach employed in Eco-SpaCE, combining ecology with ecotoxicology, provides a viable option to explore the relative contribution of contamination to the overall stress in an ecosystem. This can help environmental managers to prioritize management options, and to reduce local risks. PMID- 20005558 TI - The effect of the long term aspirin administration on the progress of atherosclerosis in apoE-/- LDLR-/- double knockout mouse. AB - We have investigated the effects of differential aspirin doses on atherogenesis. Aspirin was given to homozygous, apoE(-/-) and LDLR(-/-) double deficient mice for 12 weeks. The development of arteriosclerosis was determined morphologically by image analysis and endothelial cell function was assessed by measurement of peripheral nitric oxide (NO). METHODS: ApoE(-/-) LDLR(-/-)double knockout mice were bred and maintained with a high fat diet containing aspirin (4 and 40 mg/kg B.W. /day) for twelve weeks. The development of arteriosclerosis was monitored by estimating the total area of atherosclerotic lesions in the entire aorta. Acetylcholine-induced NO release was measured in vivo using electrochemical sensors. The expression of eNOS on the endothelial surface was determined by immuno-staining. Plasma prostaglandin F1alpha (PGF(1 alpha)), serum thromboxian B(2) (TXB(2)) and total cholesterol were measured using enzymatic assay. Bleeding time was measured by tail cut method. RESULTS: Arteriosclerosis in the 4 mg/kg/day aspirin group was decreased significantly compared with the placebo group, but not in the 40 mg/kg/day aspirin group. Acetylcholine-induced NO release was significantly depressed in the 40 mg/kg/day aspirin group. Immunochemical analysis with anti-eNOS antibody supported these findings. In the 4 mg/kg/day aspirin group, the severe suppression of PGI(2) production was not confirmed in spite of decreasing TXB(2) production, but not in the 40 mg/kg/day aspirin group. CONCLUSION: Our results suggest that endothelial dysfunction with low dose aspirin improved, reduced progression of atherosclerosis in apoE(-/-) and LDLR(-/-) double deficient mice and provides a pathophysiological basis for the beneficial effects of aspirin in atherosclerosis, and low doses appeared to be more efficient than high doses. PMID- 20005561 TI - Use of an adenosine triphosphate assay, and simultaneous staining with fluorescein diacetate and propidium iodide, to evaluate the effects of cryoprotectants on hard coral (Echinopora spp.) oocytes. AB - The objective was to examine the effects of cryoprotectants on oocytes of hard corals (Echinopora spp.) to obtain basic knowledge for cryopreservation procedures. Oocytes were exposed to various concentrations of cryoprotectants (0.25 to 5.0M) for 20 min at room temperature (25 degrees C). Two tests were used to assess ovarian follicle viability: fluorescein diacetate (FDA)+propidium iodide (PI) staining, and adenosine triphosphate (ATP) assay. Both FDA+PI staining and ATP assay indicated that cryoprotectant toxicity to oocytes increased in the order methanol, dimethyl sulfoxide (DMSO), propylene glycol (PG), and ethylene glycol (EG). The no observed effect concentrations for Echinopora spp. oocytes were 1.0, 0.5, 0.25, and 0.25 M for methanol, DMSO, PG, and EG, respectively, when assessed with FDA+PI. The ATP assay was more sensitive than FDA+PI staining (P<0.05). Oocyte viability after 1.0M methanol, DMSO, EG, or PG treatment for 20 min at room temperature assessed with FDA+PI tests and ATP assay were 88.9+/-3.1% and 72.2+/-4.4%, 66.2+/-5.0% and 23.2+/-4.9%, 58.9+/-5.4% and 1.1+/-0.7%, and 49.1+/-5.1% and 0.9+/-0.5%, respectively. We inferred that the ATP assay was a valuable measure of cellular injury after cryoprotectant incubation. The results of this study provided a basis for development of protocols to cryopreserve coral oocytes. PMID- 20005562 TI - Semen characteristics of genetically identical male cats cloned via somatic cell nucleus transfer. AB - We investigated the sperm characteristics of four cloned male cats (Felis catus) to assess their reproductive potential. Fresh and frozen-thawed sperm were assessed for motility, viability, and morphology, and their functional competence was evaluated by in vitro fertilization (IVF) of domestic cat oocytes. All fresh semen characteristics varied among cats and collection times. Sperm concentration (x 10(6)/mL) of Cat A (512+/-140, range 368 to 685) was significantly higher, whereas that of Cat C (335+/-92, range 274 to 469) was significantly lower than that of Cloned B (459+/-159, range 336 to 510) and control cats (680+/-452, range 360 to 479). After thawing, motility and progressive motility of sperm from Cat B were significantly lower than that of the other cloned and control cats. The curvilinear, straight line, and average path velocities of sperm from Cat B were significantly higher, whereas the straightness was lower, than that of the other cloned and control cats. Frozen sperm from Cats A, B, and C successfully fertilized oocytes (cleavage=74.4%, 71.4%, and 86.2%, respectively) and produced embryos that developed to the blastocyst stage after IVF/In vitro culture (IVC) (34.4%, 26.7%, and 48.0%) at frequencies similar to the cleavage rate (82.0%) and blastocyst rate (43.9%) obtained with sperm from the control male. In conclusion, seminal characteristics of cloned male cats did not differ markedly from those of our noncloned, control male cats. PMID- 20005563 TI - Strength of the bond as a predictor of marginal performance: an in vitro evaluation of contemporary adhesives. AB - OBJECTIVE: The aim of this study was to elucidate whether marginal performance (in terms of % of continuous margins) can, or not, be predicted from microTBS values. A relationship was attempted between microTBS and % of continuous margins (% CM) before and after loading of class V restorations made with five restorative systems from both etch & rinse and self-etch approach. METHODS: Resin dentin slabs and as well as class V cavities with cervical margins in dentin were prepared and restored using five adhesive systems (Scotchbond Multipurpose Plus (SMPP), James 2, Experimental Bond, Xeno III and iBond) with their corresponding restorative composites. Bond strength and percentages of continuous margins were calculated before and after thermal and mechanical loading. Statistical correlation was calculated between bond strength/marginal adaptation before loading and bond strength/marginal adaptation after loading of the same adhesives. RESULTS: Ranking of materials according to their performance was test method dependent. A low weak correlation was found between bond strength and percentages of marginal adaptation. SIGNIFICANCE: No correlation could be established between strength of the bond and percentages of continuous margins. A direct comparison of bond strength with marginal adaptation data may be misleading. PMID- 20005564 TI - Effect of element concentration on nickel release from dental alloys using a novel ion beam method. AB - OBJECTIVES: Nickel chromium is widely used as a restorative material in dentistry but its biocompatibility is of concern as there are reports of patients suffering adverse effects caused by exposure to nickel-based restorations. The aim of this work was to quantify the amount of nickel released into solution from commercially available nickel-based alloys with varying compositions and to identify the potential use of thin films in further understanding the role of chromium in reducing nickel release. METHODS: Six commercially available nickel based alloys were cast using the lost wax technique. Nickel chromium thin films were deposited onto silicon substrates by ion beam assisted physical vapor deposition. Both types of alloys were immersed into solution representative of saliva at pH 5 for 7 days. The amount of nickel released into solution was quantified by graphite-furnace atomic absorption spectrophotometry. RESULTS: The amount of nickel released from commercially available cast alloys did not correspond to the amount of nickel within the alloy. The total amount of chromium and molybdenum within the alloys proved to be of greater influence. Increasing the amount of chromium in the thin film alloys reduced the amount of nickel released but this was not linear. SIGNIFICANCE: Differences in the composition of commercial cast alloys highlighted the importance of other elements within the alloy on reducing the amount of nickel released. The use of thin film alloys can be useful in further understanding how the composition of an alloy can affect the amount of nickel released. PMID- 20005565 TI - Dentin bonding-variables related to the clinical situation and the substrate treatment. AB - The wetness of dentin surfaces, the presence of pulpal pressure, and the thickness of dentin are extremely important variables during bonding procedures, especially when testing bond strength of adhesive materials in vitro with the intention of simulating in vivo conditions. The ultimate goal of a bonded restoration is to attain an intimate adaptation of the restorative material with the dental substrate. This task is difficult to achieve as the bonding process is different for enamel and for dentin-dentin is more humid and more organic than enamel. While enamel is predominantly mineral, dentin contains a significant amount of water and organic material, mainly type I collagen. This humid and organic nature of dentin makes this hard tissue very challenging to bond to. Several other substrate-related variables may affect the clinical outcome of bonded restorations. Bonding to caries-affected dentin is hampered by its lower hardness and presence of mineral deposits in the tubules. Non-carious cervical areas contain hypermineralized dentin and denatured collagen, which is not the ideal combination for a bonding substrate. Physiological transparent root dentin forms without trauma or caries lesion as a natural part of aging. Similar to the transparent dentin observed underneath caries lesions, the tubule lumina become filled with mineral from passive chemical precipitation, making resin hybridization difficult. An increase in number of tubules with depth and, consequently, increase in dentin wetness, make bonding to deeper dentin more difficult than to superficial dentin. While the application of acidic agents open the pathway for the diffusion of monomers into the collagen network, it also facilitates the outward seepage of tubular fluid from the pulp to the dentin surface, deteriorating the bonding for some of the current adhesives. Some dentin desensitizers have shown some promise as they can block dentinal tubules to treat and prevent sensitivity and simultaneously blocking the tubular fluid from flowing to the surface. A new approach to stop the degradation of dentin-resin interfaces is the use of MMP inhibitors. Although still in an early phase of in vitro and clinical research, this method is promising. PMID- 20005566 TI - Sleep patterns and impulse control among Japanese junior high school students. AB - Adolescents with decreased impulse control exhibit behavioral problems. Lifestyles are related to impulse control. However, the relations of sleep patterns and impulse control among adolescents are unknown. Thus we examined how sleep patterns were associated with impulse control among Japanese junior high school students. Surveys were completed by a nationwide sample of 1934 students. A significant association between decreased impulse control and bedtimes after midnight was revealed. Specific lifestyle factors related to bedtimes after midnight were older age, greater numbers of hours spent watching television, lack of participation in an extracurricular activity, greater use of convenience stores, and increased attendance at cram schools. This study revealed that going to sleep after midnight was significantly related to decreased impulse control among adolescents. Data about specific lifestyle factors related to going to sleep after midnight should be useful in preventing those behaviors demonstrated by school children that derive from decreased impulse control. PMID- 20005567 TI - Is toxicological pathology characterised by a loss of system complexity? AB - Physiological function emerges from complex biomolecular interactions (e.g., protein-protein) and control mechanisms that enable animals to respond and adapt to changes in their environment. Cell injury and pathology induced by pollutants and other stressors appears to involve the gradual and progressive dysfunction of complex biomolecular interactions, resulting in loss of integrated physiological interactions and homeostasis leading to a reduced capacity to respond effectively to stress. In order to determine whether complexity can be used as an indicator of health, the hypothesis that pathology involves a loss of biological complexity has been tested using a generic physiological interaction network. System complexity was evaluated using Eulerian cycles and connectedness (connectance%) for estimating topological complexity and application of network theory (i.e., analysis of scale-free networks and network diameter). The complexity of the whole system increases when sub-systems, such as detoxication and anti-oxidant protective processes, augmented autophagy, protein degradation and induction of stress proteins, are up-regulated and start to interact significantly as part of a response to low-level stress (i.e., biphasic or hormetic response). However, with increasing severity of stress, cell injury and higher-level functional impairment lead to physiological dysfunction and breakdown of the whole interaction network with consequent loss of complexity. In summary for the model described here, network and graph theory appear to provide a mathematical formalism that can facilitate the system-level interpretation of health and dysfunction in living cells. PMID- 20005568 TI - Polycyclic aromatic hydrocarbons and petroleum biomarkers in Sao Sebastiao Channel, Brazil: assessment of petroleum contamination. AB - Polycyclic aromatic hydrocarbons (PAHs) and non-aromatic hydrocarbons (NAHs), including n-alkanes, isoprenoids and petroleum biomarkers (terpanes, hopanes, steranes and diasteranes), were quantified by gas chromatography with flame ionization and mass spectrometer detectors in sediment samples collected from the Sao Sebastiao Channel (SSC), Brazil, where the largest Brazilian maritime petroleum terminal is located. The concentrations of total PAHs, total n-alkanes and petroleum biomarkers ranged from below the detection limits to 370ngg(-1), 28microgg(-1), 2200ngg(-1) (dry weight), respectively. The analysis of PAH distribution suggested combustion sources of PAHs as the main input for these compounds with smaller amount from petroleum contamination. The distribution of petroleum biomarkers undoubtedly demonstrated petroleum as a source of anthropogenic contamination throughout the region. The assessment of petrogenic sources of contamination in marine sediment is more challenging if only PAH analysis were carried out, which demonstrates that more stable hydrocarbons such as petroleum biomarkers are useful for investigating potential presence of petroleum. PMID- 20005569 TI - Adult cell therapy for brain neuronal damages and the role of tissue engineering. AB - No long term effective treatments are currently available for brain neurological disorders such as stroke/cerebral ischemia, traumatic brain injury and neurodegenerative disorders. Cell therapy is a promising strategy, although alternatives to embryonic/foetal cells are required to overcome ethical, tissue availability and graft rejection concerns. Adult cells may be easily isolated from the patient body, therefore permitting autologous grafts to be performed. Here, we describe the use of adult neural stem cells, adrenal chromaffin cells and retinal pigment epithelium cells for brain therapy, with a special emphasis on mesenchymal stromal cells. However, major problems like cell survival, control of differentiation and engraftment remain and may be overcome using a tissue engineering strategy, which provides a 3D support to grafted cells improving their survival. New developments, such as the biomimetic approach which combines the use of scaffolds with extracellular matrix molecules, may improve the control of cell proliferation, survival, migration, differentiation and engraftment in vivo. Therefore, we later discuss scaffold properties required for brain cell therapy as well as new tissue engineering advances that may be implemented in combination with adult cells for brain therapy. Finally, we describe an approach developed in our laboratory to repair/protect lesioned tissues: the pharmacologically active microcarriers. PMID- 20005570 TI - Cullins in human intra-uterine growth restriction: expressional and epigenetic alterations. AB - Intra-uterine growth restriction (IUGR) is defined by a restriction of fetal growth during gestation. It is a prevalent significant public health problem that jeopardizes neonatal health but also that can have deleterious consequences later in adult life. Cullins constitute a family of seven proteins involved in cell scaffold and in selective proteolysis via the ubiquitin-proteasome system. Most Cullins are critical for early embryonic development and mutations in some Cullin genes have been identified in human syndromes including growth retardation. Our work hypothesis is that Cullins, particularly CUL4B and CUL7, are involved in placental diseases and especially in IUGR. Thus, expression of Cullins and their cofactors was analyzed in normal and pathological placentas. We show that they present a constant significant over-expression in IUGR placentas, whose extent is dependent on the position of the interrogated fragment along the cDNAs, suggesting the existence of different isoforms of the genes. Particularly, the CUL7 gene is up-regulated up to 10 times in IUGR and 15 times in preeclampsia associated with IUGR. The expression of cofactors of Cullins participating to functional complexes has also been evaluated and showed a similar significant increase in IUGR. Promoters of Cullin genes appeared to be under the control of the SP1 transcription factor. Finally, methylation levels of the CUL7 promoter in placental tissues are modulated according to the pathological conditions, with a significant hypomethylation in IUGR. These results concur to pinpoint the Cullin family as a new set of markers of IUGR. PMID- 20005571 TI - Feto-maternal interactions in pregnancies: placental microparticles activate peripheral blood monocytes. AB - Normal pregnancy is associated with a systemic maternal inflammatory reaction, including the activation of peripheral blood monocytes. This reaction is exaggerated in pre-eclampsia, a severe placenta-dependent disorder of pregnancy specific to humans. It has been suggested that placental syncytiotrophoblast membrane microparticles (STBM), which are released into the peripheral blood, may contribute to the maternal response. The aim of this study was to investigate the inflammatory properties of STBM generated by four different approaches on primary human monocytes in vitro. Cellular viability, phenotype and functional response were analysed. STBM isolated by mechanical dissection and STBM generated from villous explant cultures incubated in hypoxic conditions had only minor influences on the monocytic phenotype and failed to induce a proinflammatory response. By contrast, STBM washed from the maternal side of a placental cotyledon and STBM shed by explants cultured in air up-regulated cell surface expression of the adhesion molecule CD54 and induced the production of interleukin (IL)-8, IL-6 and IL-1beta. Cytokine production was time- and dose dependent. Our study, therefore, suggests that monocyte activation in normal pregnancy and pre-eclampsia may be induced by STBM released by the placenta. The higher amounts of STBM circulating in maternal blood in pre-eclampsia might lead to the excessive maternal inflammatory reaction. PMID- 20005572 TI - Short- and long-term changes in gene expression mediated by the activation of TLR9. AB - CpG DNA binds to Toll-like receptor 9 to stimulate a strong innate immune response. The magnitude, duration and scope of CpG-induced changes in gene expression are incompletely understood despite extensive studies of TLR9 mediated signal transduction pathways. In particular, the prolonged effects of CpG DNA on gene activation have not been investigated despite evidence that a single dose of CpG DNA alters immune reactivity for several weeks. This study used gene expression analysis to monitor changes in mRNA levels for 14 days, and identified the genes, pathways and functional groups triggered in vivo following CpG DNA administration. Two discrete peaks of gene activation (at 3h and 5 days) were observed after CpG injection. Both the behavior and function of genes activated during the second peak differed from those triggered shortly after CpG administration. Initial gene up-regulation corresponded to a period when TLR9 ligation stimulated genes functionally associated with the generation of innate and adaptive immune responses (e.g. the NF-kappaB and B-cell receptor pathways). The second peak reflected processes associated with cell division (e.g. cell cycle and DNA replication and repair). The complex bimodal pattern of gene expression elicited by CpG DNA administration provides novel insights into the long-term effects of TLR9 engagement on genes associated with immunity and cell proliferation. PMID- 20005573 TI - Role of the hepatocyte growth factor gene in refractive error. AB - OBJECTIVE: Refractive errors such as myopia and hypermetropia are among the leading causes of visual impairment worldwide. Several genetic loci have been associated with myopia but none to date have been reported for hypermetropia. We investigated the hepatocyte growth factor (HGF) as a candidate gene influencing these 2 refractive error states. DESIGN: Case-control study. PARTICIPANTS: A total of 551 individuals (193 males, 358 females; mean age, 55.41+/-12.65 years) including 117 individuals with high myopia +2.00 D) were included in the analysis from 3 different Australian population cohorts (The Genes in Myopia Study, the Blue Mountains Eye Study, and the Melbourne Visual impairment project). METHODS: Genotyping of 9 tag single nucleotide polymorphisms (SNPs) that encompassed the entire HGF gene and its associated sequences as well as 6 additional SNPs identified through DNA resequencing was undertaken. MAIN OUTCOME MEASURES: Genetic association with refraction. RESULTS: After correction for multiple testing, the SNPs rs12536657 (odds ratio [OR], 5.53; 95% confidence interval [CI], 1.14-26.76) and rs5745718 (OR, 2.24; 95% CI, 1.30-3.85) showed significant association with hypermetropia. Whereas the SNPs rs1743 (OR, 2.02; 95% CI, 1.19-3.43; P = .009), rs4732402 (OR, 2.03; 95% CI, 1.23-3.36; P = 0.005), rs12536657 (OR, 2.38; 95% CI, 1.40-4.05; P = 0.001), rs10272030 (OR, 2.22; 95% CI, 1.31-3.75; P = 0.003), and rs9642131 (OR, 2.44; 95% CI, 1.43-4.14; P = 0.001) showed significant association with low/moderate myopia. CONCLUSIONS: These findings present the HGF gene as the first gene significantly associated with hypermetropia as well as providing evidence of significant association with myopia in a second ethnic population. In addition, it provides insights into the important biological mechanisms that regulate human ocular development (emmetropization), which are currently poorly understood. PMID- 20005575 TI - Cytokine production by peripheral blood mononuclear cells of women with a history of preterm birth. AB - Preterm birth is associated with elevated production of pro-inflammatory cytokines such as TNFalpha at the maternal-fetal interface. Previous studies have suggested that women with a history of preterm birth produce aberrantly strong inflammatory responses to bacterial lipopolysaccharide (LPS). However many intrauterine infections in women are associated with pathogens including Ureaplasma urealyticum, Mycoplasma hominis and Streptococcus agalactiae (group B streptococcus) that contain pro-inflammatory factors other than LPS. We evaluated whether peripheral blood leukocytes from women with a history of preterm birth produce elevated amounts of TNFalpha upon stimulation with pathogens associated with preterm birth and if pre-treatment with aspirin, an anti-inflammatory medication, decreases the ex vivo production of this cytokine. Heat-killed bacteria elicited increased TNFalpha production from leukocytes in a dose dependent manner, but no differences in TNFalpha production between leukocytes from women with preterm birth and control women with term birth were detected. In women who consumed aspirin each day for one week, TNFalpha production was increased in leukocytes from control women stimulated with Escherichia coli and U. urealyticum, but was reduced or unchanged in leukocytes from women with preterm birth. Similar trends were observed for a subset of samples stimulated with U. urealyticum and assayed for IL-6, IL-10, IL-1beta and TNFalpha by bead array. We conclude that leukocytes from women with a history of preterm birth do not have elevated pro-inflammatory responses to pathogens, and that reproductive history is associated with different effects of aspirin on pro-inflammatory cytokine production. PMID- 20005574 TI - Binding of alpha-synuclein with Fe(III) and with Fe(II) and biological implications of the resultant complexes. AB - Parkinson's disease (PD) is hallmarked by the abnormal intracellular inclusions (Lewy bodies or LBs) in dopaminergic cells. Amyloidogenic protein alpha-synuclein (alpha-syn) and iron (including both Fe(III) and Fe(II)) are both found to be present in LBs. The interaction between iron and alpha-syn might have important biological relevance to PD etiology. Previously, a moderate binding affinity between alpha-syn and Fe(II) (5.8x10(3)M(-1)) has been measured, but studies on the binding between alpha-syn and Fe(III) have not been reported. In this work, electrospray mass spectrometry (ES-MS), cyclic voltammetry (CV), and fluorescence spectroscopy were used to study the binding between alpha-syn and Fe(II) and the redox property of the resultant alpha-syn-Fe(II) complex. The complex is of a 1:1 stoichiometry and can be readily oxidized electrochemically and chemically (by O(2)) to the putative alpha-syn-Fe(III) complex, with H(2)O(2) as a co-product. The reduction potential was estimated to be 0.025V vs. Ag/AgCl, which represents a shift by -0.550V vs. the standard reduction potential of the free Fe(III)/Fe(II) couple. Such a shift allows a binding constant between alpha-syn and Fe(III), 1.2x10(13)M(-1), to be deduced. Despite the relatively high binding affinity, alpha-syn-Fe(III) generated from the oxidation of alpha-syn-Fe(II) still dissociates due to the stronger tendency of Fe(III) to hydrolyze to Fe(OH)(3) and/or ferrihydrite gel. The roles of alpha-syn and its interaction with Fe(III) and/or Fe(II) are discussed in the context of oxidative stress, metal-catalyzed alpha-syn aggregation, and iron transfer processes. PMID- 20005576 TI - Differences in genetic background influence the induction of innate and acquired immune responses in chickens depending on the virulence of the infecting infectious bursal disease virus (IBDV) strain. AB - Previous studies and field observations have suggested that genetic background influences infectious bursal disease virus (IBDV) pathogenesis. However, the influence of the virulence of the infecting IBDV strain and the mechanisms underlying the differences in susceptibility are not known. In the present study IBDV pathogenesis was compared between specific-pathogen-free layer-type (LT) chickens, which are the most susceptible chicken for IBDV and have been used as the model for pathogenesis studies, and broiler-type (BT) chickens, which are known to be less susceptible to clinical infectious bursal disease (IBD). The innate and acquired immune responses were investigated after inoculation of an intermediate (i), virulent (v) or very virulent (vv) strain of IBDV. IBDV pathogenesis was comparable among genetic backgrounds after infection with iIBDV. After infection with vIBDV and vvIBDV, LT birds showed severe clinical disease and mortality, higher bursal lesion scores and IBDV-antigen load relative to BT birds. Circulating cytokine induction varied significantly in both timing and quantity between LT and BT birds and among virus strains (P<0.05). Evaluation of different immune cell populations by flow-cytometric analysis in the bursa of Fabricius provided circumstantial evidence of a stronger local T cell response in BT birds vs. LT birds after infection with the virulent strain. On the other hand, LT birds showed a more significant increase in circulating macrophage derived immune mediators such as total interferon (IFN) and serum nitrite than BT birds on days 2 and 3 post-vIBDV infection (P<0.05). Stronger stimulation of innate immune reactions especially after vIBDV infection in the early phase may lead to faster and more severe lesion development accompanied by clinical disease and death in LT chickens relative to BT chickens. Interestingly, no significant differences were seen between genetic backgrounds in induction of the IBDV specific humoral response: timing of IBDV-antibody induction and antibody levels were comparable between BT and LT birds. This study clearly demonstrates a significant influence of chickens' genetic background on disease outcome. The difference between backgrounds in IBDV susceptibility is further influenced by the virulence of the infecting virus strain. PMID- 20005577 TI - Multiple bovine FcgammaRIIb sub-isoforms generated by alternative splicing. AB - Receptors for the Fc portion of immunoglobulin molecules (FcR) provide an important and vital link between circulating antibody and cellular effector functions. These receptors have been well characterized in human and murine species, however few of these receptors have been investigated in livestock. FcgammaRII (CD32) is an FcR previously shown in mice and humans to exist in multiple isoforms, both activating (FcgammaRIIa, FcgammaRIIc) and inhibitory (FcgammaRIIb), on a wide variety of cells including B cells, T cells, dendritic cells, monocytes, macrophages and platelets. On B cells, FcgammaRIIb acts to suppress cell activation and immunoglobulin production by means of an intracellular immunoreceptor tyrosine-based inhibitory motif signaling domain. Two sub-isoforms of FcgammaRIIb, designated b1 and b2, distinguished by the inclusion of an additional cytoplasmic exon in the b1 form, have been demonstrated in humans and mice, whereas only one sequence corresponding to the human and mouse b2 isoform has been identified in cattle. In this study, the expression profile of FcgammaRIIb in bovine blood mononuclear cells was characterized by collecting blood samples from mature cattle of dairy and beef breeds, and determining their FcgammaRIIb mRNA expression profile by RT-PCR. Analysis revealed the presence of two uncharacterized bovine FcgammaRIIb transcripts in addition to the single previously published transcript. Analysis of the first unknown transcript revealed high homology with published human and murine FcgammaRIIb1 sequences. This transcript was present in all cell types examined, with little variation in primary sequence between individuals or among breeds. The second unknown sequence was found to be homologous to the murine FcgammaRIIb3 (IgG-binding protein or soluble FcgammaR in humans) sequence. This transcript appears to have a much more limited expression profile, which may indicate that expression varies with the cellular activation-state of the cell. These results indicate that cattle, like humans and mice, express multiple sub isoforms of FcgammaRIIb. These findings add further complexity to the regulation of IgG-mediated immunity and provide new insight into the role Fc receptors play in antigen acquisition and presentation in cattle. PMID- 20005578 TI - An epitope from Acanthamoeba castellanii that cross-react with proteolipid protein 139-151-reactive T cells induces autoimmune encephalomyelitis in SJL mice. AB - We report here that an epitope (aa, 83-95) derived from Acanthamoeba castellanii (ACA) induces clinical signs of experimental autoimmune encephalomyelitis (EAE) in SJL/J mice reminiscent of the disease induced with myelin proteolipid protein (PLP) 139-151. By using IA(s)/tetramers, we demonstrate that both ACA 83-95 and PLP 139-151 generate antigen-specific cross-reactive CD4 T cells and the T cells secrete identical patterns of cytokines and induce EAE with a similar severity. These results may provide insights into the pathogenesis of multiple sclerosis and ACA-induced granulomatous encephalitis. PMID- 20005579 TI - Surgical management of intraoral ranulas in children: an analysis of 17 pediatric cases. AB - OBJECTIVE: The preferred treatment of pediatric intraoral ranulas remains controversial. We present our experience with ranulas at the Gyeongsang National University Hospital and review the literature. METHODS: The study involved 17 children under 15 years of age who visited our hospital from 2001 to 2008 and were diagnosed with a ranula exceeding 2 cm in diameter. The age and gender of the patients, the surgical procedures, presurgical observation period, postsurgical follow-up period, and complications of each case were determined. The unruptured specimens were all subjected to detailed pathological analysis. RESULTS: The patients were on average 9.3 years and there were 10 girls and 7 boys. Spontaneous resolution was not detected in any of the cases during the presurgical observation period. Indeed, in two cases, the ranula had increased in size. The ranula and sublingual gland (SG) were resected in all cases. The average operation time was 1h. Recurrence and complications were not detected in any of the cases. Pathological analyses revealed that there was no communication of the ranula with the SG in any of the cases. CONCLUSIONS: Our experiences suggest that the presurgical observation period need not be longer than 3 months and that the resection of ranulas along with the ipsilateral SG is a safe and effective primary treatment for symptomatic pediatric intraoral ranulas that exceed 2cm in diameter. PMID- 20005580 TI - The prairie vole: an emerging model organism for understanding the social brain. AB - Unlike most mammalian species, the prairie vole is highly affiliative, forms enduring social bonds between mates and displays biparental behavior. Over two decades of research on this species has enhanced our understanding of the neurobiological basis not only of monogamy, social attachment and nurturing behaviors but also other aspects of social cognition. Because social cognitive deficits are hallmarks of many psychiatric disorders, discoveries made in prairie voles can direct novel treatment strategies for disorders such as autism spectrum disorder and schizophrenia. With the ongoing development of molecular, genetic and genomic tools for this species, prairie voles will likely maintain their current trajectory becoming an unprecedented model organism for basic and translational research focusing on the biology of the social brain. PMID- 20005581 TI - Catalytic and immunochemical detection of hepatic and extrahepatic microsomal cytochrome P450 1A1 (CYP1A1) in white-sided dolphin (Lagenorhynchus acutus). AB - We have characterized microsomal systems and measured the levels of microsomal cytochrome P450 1A1 (CYP1A1) and ethoxyresorufin-O-deethylase (EROD) activity in multiple internal organs of male and female white-sided dolphin (Lagenorhynchus acutus) from the northwest Atlantic Ocean. Internal organs were sampled within 24h of death, sometimes in a period of hours, collection times which are significantly less than usually seen for marine mammals. Tissue autolysis, as assessed by histological analysis of liver, was minimal to none in all individuals. Total P420 did not correlate with time from death to sampling, suggesting that it is a poor indicator of P450 degradation in cetacean tissues where perfusion is not practical. The total hepatic microsomal P450 content, cytochrome b5 content, and NADPH-cytochrome c (P450) reductase (CPR) activity averaged 0.29nmolmg(-1), 0.12nmolmg(-1), and 238nmolmg(-1)min(-1), respectively. Microsomal CPR activity in liver was higher than that in lung and kidney, and was higher than that reported in liver of most other cetacean species. Immunodetected CYP1A1 content was low in all organs, less than 3pmolesCYP1A equivalentsmg(-1). EROD activity ranged from 9 to 376pmolesmg(-1)min(-1) and was greater in liver than in other tissues. Hepatic microsomal EROD activity and CYP1A1 content did not correlate. However, hepatic EROD activity, but not CYP1A1 protein content, was well correlated with both total PCB and Sigmamono-ortho PCB concentrations in blubber. Length, as a proxy for age, did not correlate with hepatic EROD activity or CYP1A1 protein levels, and sex did not influence the relationship between EROD and contaminant concentrations. We cannot easily control for the extent of tissue degradation in cetacean studies nor do we have a complete history of these animals. Therefore, other factors such as degradation or hormonal state may have a role in the observed relationships. Yet, as in other mammals, hepatic tissues appear to be a major site of CYP1A1 expression and probably of biotransformation of CYP1A substrates in white-sided dolphin. The expression of an EROD catalyst in liver likely reflects induction by PCBs, but the P450 enzyme catalyzing hepatic EROD activity in these whales may not be CYP1A1. PMID- 20005582 TI - Assessing the effects of exposure timing on biomarker expression using 17beta estradiol. AB - Temporal and spatial variability in estrogenicity has been documented for many treated wastewater effluents with the consequences of this variability on the expression of biomarkers of endocrine disruption being largely unknown. Laboratory exposure studies usually utilize constant exposure concentrations which may produce biological effects that differ from those observed in organisms exposed in natural environments. In this study, we investigated the effects of differential timing of exposures with 17beta-estradiol (E2) on a range of fathead minnow biomarkers to simulate diverse environmentally relevant exposure profiles. Two 21-day, replicate experiments were performed exposing mature male fathead minnows to E2 at time-weighted mean concentrations (similar average exposure to the contaminant during the 21-day exposure period; 17ng E2/L experiment 1; 12ng E2/L experiment 2) comparable to E2 equivalency values (EEQ) reported for several anthropogenically altered environments. A comparable time-weighted mean concentration of E2 was applied to five treatments which varied in the daily application schema: E2 was either applied at a steady rate (ST), in a gradual decreasing concentration (HI), a gradual increasing concentration (LO), applied intermittently (IN), or at a randomly varying concentration (VA). We assessed a range of widely used physiological (vitellogenin mRNA induction and plasma concentrations), anatomical (body and organ indices, secondary sex characteristics, and histopathology), and behavioral (nest holding) biomarkers reported to change following exposure to endocrine active compounds (EACs). All treatments responded with a rise in plasma vitellogenin concentration when compared with the ethanol carrier control. Predicatively, vitellogenin mRNA induction, which tracked closely with plasma vitellogenin concentrations in most treatments was not elevated in the HI treatment, presumably due to the lack of E2 exposure immediately prior to analysis. The ability of treatment male fish to hold nest sites in direct competition with control males was sensitive to E2 exposure and did yield statistically significant differences between treatments and carrier control. Other biological endpoints assessed in this study (organosomatic indices, secondary sex characteristics) varied little between treatments and controls. This study indicates that a broad suite of endpoints is necessary to fully assess the biological consequences of fish exposure to estrogens and that for at least field studies, a combination of vitellogenin mRNA and plasma vitellogenin analysis are most promising in deciphering exposure histories of wild-caught and caged fishes. PMID- 20005583 TI - Receptors and aging: dedicated to the memory of Paul Ehrlich for the 100th anniversary of his Nobel Prize. AB - The initiation and evolution of the receptor concept and its application in pharmacology can be traced back to Paul Ehrlich's original experiments. Since several decades the receptor concept is in the foreground of cell biology and pharmacology. We present here a short reminder of Ehrlich's concepts on receptor action, its evolution and modifications as a result of increasing life expectancy of human societies. Results obtained by several teams on the age-dependent modifications of receptor function are reviewed with special emphasis on the age dependent loss of receptors and of uncoupling of receptors from their intracellular transmission pathway. As a special example we summarize our results on the elastin receptor and its age-dependent modifications. These modifications result in the loss of the physiologically helpful functions mediated by this receptor, such as vasodilation by coupling with the inducible nitric oxide synthase (iNOS)-inhibition of cellular cholesterol synthesis and modulation of free radical production by inhibition of the guanine nucleotide binding protein (Gi protein)-mediated transmission pathway. Only the harmful effects such as free radical release and up-regulation of elastase production remain in "old" cells. The age-dependent modifications of receptor function play an important role in the increasing frequency and severity of age-related diseases such as athero arteriosclerosis and emphysema as well as the loss of hormone- and drug actions. These processes and their inhibition or correction represent a new challenge for cellular pharmacology. PMID- 20005584 TI - Training in elder abuse: the experience of higher specialist trainees in Geriatric Medicine in the UK. AB - Little is known about elder abuse training in post-graduate medical education in the United Kingdom. A questionnaire survey was conducted among Specialist Registrars (SpRs) in Geriatric Medicine attending a single training day within five UK Post-graduate Deaneries to assess self-report experience of elder abuse training during their post-graduate medical education. The questionnaire included quantity, quality and self-report confidence in dealing with elder abuse issues. A total of 112 questionnaires were returned (response rate 100%, representing 78.9% of eligible trainees within these five deaneries). The majority of the participants rated low scores for both quantity and quality of training they received. This finding was consistent across all 5 years of training with no significant difference in rating scores in more experienced (years 3-5) SpRs. The majority (62.5%) of final year SpRs reported feeling inadequately prepared to deal with cases of elder abuse. The results of our survey suggest that the provision and quality of training regarding elder abuse is poor for trainees in Geriatric Medicine. The majority of SpRs approaching the completion of their training felt ill-prepared in managing such cases. Our study highlights the requirement of structured approach in elder abuse training for trainees in Geriatric Medicine. PMID- 20005585 TI - Prevalence of musculoskeletal pain in an elderly Korean population: results from the Korean Longitudinal Study on Health and Aging (KLoSHA). AB - The aim of this study is to estimate the prevalence of musculoskeletal (MSK) pain in the upper extremities (UE), low back, and lower extremities (LE) in an elderly population, and to identify related factors. In this cross-sectional cohort study, 1118 Korean elderly subjects were randomly selected from residents aged >=65 years living in an urban city of Korea. The study data included presence of MSK pain, educational levels, activity levels over a 24h, monthly income, body mass indices and presence of depression. Estimated age- and gender-standardized prevalences of UE, low back, and LE pain were 62.6%, 72.6%, and 45.7%, respectively. The prevalence of LE pain increased with age, whereas those of UE pain and back pain did not. By multivariate analysis, a female gender and a low income were found to be significantly associated with pain at all sites, and obesity was related with low back and LE pain. Furthermore, an uneducated state was found to be associated with LE pain, and major depressive disorder to be obviously related to UE and LE pain. MSK pain was identified to be a common problem in the elderly Korean population, and a female gender was consistently associated with MSK pain. PMID- 20005586 TI - Coexisting vasospastic angina and undiagnosed Brugada syndrome resulting in cardiac arrest. AB - The coexistence of Brugada Syndrome and resting vasospastic angina resulting in cardiac arrest is rare. We describe a 64 year-old man presenting with cardiac arrest and vasospastic angina with diagnostic criteria of symptomatic Brugada Syndrome. Recognition of the coexistence of these potentially fatal conditions has important therapeutic implications when using calcium channel antagonists and may shed light on the mechanisms of coronary spasm. A common pathogenesis, such as a common underlying channelopathy, may explain its coexistence with Brugada Syndrome. PMID- 20005587 TI - Susceptibility to paratuberculosis infection in cattle is associated with single nucleotide polymorphisms in Toll-like receptor 2 which modulate immune responses against Mycobacterium avium subspecies paratuberculosis. AB - Paratuberculosis is a chronic intestinal infection in ruminants, caused by Mycobacterium avium subspecies paratuberculosis (Map). To study the role of host genetics in disease susceptibility, the Toll-like receptor 2 (TLR2) gene, selected based on its potential role in immunity to mycobacterial infections, was analyzed for single nucleotide polymorphisms (SNP) and their potential association with disease. For SNP discovery and to study SNP association with disease, a case-control study including 24 cows from farms with paratuberculosis was conducted. Sequence analysis of the TLR2 genes from 12 paratuberculosis infected animals and 12 age-matched healthy herd mates revealed 21 different SNP. The TLR2-1903 T/C SNP was significantly associated with resistance to Map. This and four additional TLR2 SNP were studied in a subsequent observational field study with 553 cows from farms with paratuberculosis. The allelic distribution of the TLR2-1903 T/C SNP was confirmed to be significantly different between the infected and non-infected animals. For the TLR2-1903 T/C SNP the odds ratio was calculated, and similar to the dominance model in the association study, the CT and CC genotypes were compared to the TT genotype. Cows with the TLR2-1903 T/C mutation (i.e., the CT and CC genotypes) were at 1.7 (95% CI: 1.2, 2.8) times the odds of being Map-infected compared to cows with the TT genotype. In in vitro functional assays, monocyte-derived macrophages from animals with a TLR2-1903 TT genotype produced more IL12p40 and IL1beta when stimulated with Map compared to cells derived from TLR2-1903 CT and CC genotypes. Also, T cell proliferative responses to mycobacterial antigens were higher in animals with a TLR2-1903 TT genotype. In conclusion, we have found a significant association between SNP TLR2 1903 T/C in the bovine TLR2 gene and bovine paratuberculosis infection. This SNP and other genetic markers could be useful in marker-assisted breeding strategies as an additional tool in paratuberculosis control strategies. In addition, the functional studies suggest that genetic polymorphisms in bovine TLR2 which result in higher macrophage activity may contribute to enhanced T cell activation and a lower susceptibility to paratuberculosis in cattle. PMID- 20005588 TI - Physician incentives: cure versus prevention. AB - This paper distinguishes between two scenarios for the physician-patient encounter. In the cure scenario, the patient does not know whether a loss can be recovered. In the prevention scenario, the patient faces a threat but does not know whether this threat is real enough to justify preventive action. The patient wants to induce the physician both to give an accurate diagnosis and to put appropriate effort into cure or prevention. It is shown that in the cure scenario, a contingent fee solves both these incentive problems. In the prevention scenario, however, putting up with low effort makes it easier to get an accurate diagnosis, and the use of contingent fees should be limited. These results are interpreted as providing a rationale for observed exceptions to legal and ethical restrictions on the use of contingent fees. Indeed, such exceptions are often granted for cases that fit the cure scenario. PMID- 20005589 TI - Exploring microbial diversity for biotechnology: the way forward. AB - Environmental microbes are immensely diverse and have numerous metabolic activities and products that could have industrial applications. However, >99% of environmental microbes cannot be cultured under current laboratory conditions, leaving their potential largely untapped. Metagenomic approaches have been used successfully in recent years to obtain novel microbial products from uncultured microorganisms. The activity, efficiency and stability of these novel enzymes can be further improved by the application of nanotechnology. Here, I highlight the approaches that can be used to obtain efficient microbial products from the uncultivable majority. I propose that a multidisciplinary approach combining different technologies including metagenomics and nanotechnology is the way forward for tapping the real potential of microbial metabolism for applications in biotechnology. PMID- 20005590 TI - I-125 seed planning: an alternative method of urethra definition. AB - BACKGROUND AND PURPOSE: To investigate the use of aerated aqueous gel rather than a catheter to define the urethra during permanent I-125 seed implant planning. MATERIALS AND METHODS: Twenty patients were treated between September 2007 and March 2008, each having two sequential volume studies: one visualizing the urethra with a catheter and the other using aerated gel. Two individually optimised plans were produced for each patient: one from the gel and the other from the catheter image set, and the plans were analysed dosimetrically. The plans were also interchanged (putting gel plan onto catheter image set and vice versa), and dose homogeneity within a slice was investigated. Three patients had MRI post-implant to check urethral position and dosimetry. RESULTS: The urethra appears larger when defined with gel rather than when defined with a catheter, with volumes of 0.9+/-0.3 [range: 0.6-1.6]cc and 0.7+/-0.2 [range: 0.4-1.0]cc, respectively. Catheter plans appear dosimetrically slightly preferable to gel plans with V(100) prostate being 99.7+/-0.2 [range: 99.3-100.0]% and 99.5+/-0.5 [range: 98.12-99.9]% for catheter and gel, respectively (p=0.048). The urethra appears to receive a higher dose when defined with gel with V(150) being 0.5+/ 0.7 [range: 0-2.8]% and 0.2+/-0.2 [range: 0-2.6]% for gel- and catheter-defined urethras. Seed density, COIN and number of seeds were almost equivalent. Statistically, only V(100) prostate and D(90) prostate are significantly different. Putting the catheter plan on the gel image set shows a significant increase in urethral dose with V(150) urethra increasing significantly from 0.2+/ 0.2 [range: 0-0.6]% from the catheter plan to 15.3+/-11.9 [range: 0.8-47.0]% for the catheter plan on gel image set (p<0.001). D(90) for the inner core of gel and catheter plans was 193.9+/-6.7 [range: 180.3-202.3]Gy and 198.7+/-4.7 [range: 190.7-211.5]Gy (p=0.023). Gel plans are cooler centrally and less homogeneous, which could be counteracted by increasing seed activity and/or prescription dose, though the clinical significance of this should be investigated before implementation. CONCLUSIONS: This investigation has shown that the urethral position is distorted by the presence of a urinary catheter. The dosimetry may be different from that planned if this is not taken into account. PMID- 20005591 TI - Radiocontrast media affect radiation-induced DNA damage repair in vitro and in vivo by affecting Akt signalling. AB - PURPOSE: The study was performed to investigate cytogenetic effects of ionic and non-ionic radiocontrast media (RCM) meglumine, iohexol alone and in combination with irradiation in mouse bone marrow cells in vivo and in vitro. MATERIALS AND METHODS: Micronuclei assay was performed in bone marrow cells (BMC) of Balb/C mice intraperitoneally injected with RCM in the presence or absence of whole-body irradiation of 50 mGy. DNA repair (NHEJ) signalling and efficiency were analyzed by Western blot and gammaH2AX-foci assay in normal fibroblast HSF-7 and HUVEC cells. RESULTS: Both compounds reduced proliferation of BMC significantly. Concentrations of 0.5, 1 and 2 ml/kg meglumine or iohexol significantly enhanced the frequency of micronucleated polychromatic erythrocytes (MnPCEs) at all doses of meglumine (p<0.01) and 2 ml/kg of iohexol (p<0.05). Combined with irradiation meglumine at 0.5 and 1 ml/kg led to a higher frequency of MnPCEs than iohexol/IR (p<0.05). Meglumine induced DNA-double strand breaks (DNA-DSB) in non-irradiated HSF and strongly increased residual DNA-DSB within 10 min to 24h after irradiation with 200 or 400 mGy (p<0.001). Iohexol did not induce DNA-DSB but blocked repair of radiation-induced DNA-DSB significantly (p<0.05). Meglumine blocked IR-induced Akt phosphorylation, phosphorylation of DNA-PKcs (S2056, T2609) and ATM (S1981). Iohexol only blocked phosphorylation of Akt and DNA-PKcs at S2056. CONCLUSION: RCM result in clastogenic effects through interference intracellular signalling cascades involved in the regulation of non-homologous end-joining repair of DNA-DSB. PMID- 20005592 TI - Susceptibility to tetracycline and erythromycin of Lactobacillus paracasei strains isolated from traditional Italian fermented foods. AB - The aim of this study was to evaluate the susceptibility of 197 isolates of Lactobacillus paracasei, isolated from Italian fermented products coming from different geographical areas, to tetracycline and erythromycin, two antimicrobials widely used in clinical and animal therapy. Isolation media were supplemented with antibiotics according to the microbiological breakpoints (BPs) defined by European Food Safety Authority (EFSA). Isolates were identified at the species level and were typed by rep-PCR using the (GTG)(5) primer. A total of 121 genotypically different strains were detected and their phenotypic antimicrobial resistance to tetracycline and erythromycin was determined as the minimum inhibitory concentration (MIC) using the broth microdilution method. The presence of the genes ermB, ermC and tetL, tetM, tetS, tetW, in the phenotypically resistant isolates was investigated by PCR. Tetracycline induction of tetM expression on representative resistant strains, grown in medium either lacking or containing the antibiotic, was also analyzed by RT-PCR. Among the 121 tested strains, 77.7% were susceptible to tetracycline (MICor=1024 microg/ml) (Erm(R)). The tetM and ermB genes were the most frequently detected in the Tet(R) and/or Erm(R) strains. The tetM expression was induced by antibiotic addition to the growth medium. Our study confirmed that L. paracasei is quite sensitive to tetracycline and erythromycin, but the high level of resistance of Erm(R) strains suggested that acquired resistance took place. Further investigations are required to analyze whether the genes identified in L. paracasei isolates might be horizontally transferred to other species. Since "commensal" bacteria, which L. paracasei belongs to, may play an active role in the spreading of antibiotic resistance, a series of measures inspired from a principle of precaution should be taken before they are used as commercial starters or probiotic cultures in food products, complemented by a more prudent use of antibiotics in agriculture, veterinary, and human medicine. PMID- 20005593 TI - Prevalence of overweight and obesity in young adults aged 20-40 years in North India (Kashmir Valley). AB - We conducted a survey to know the prevalence of diabetes and obesity in young adult (20-40 years) men and non-pregnant-women in Kashmir Valley (India); prevalence of latter is presented here. The age-adjusted prevalence of overweight obesity was 16.3% and that of obesity 5.1%; the relationship of overweight obesity with glucose intolerance is discussed. PMID- 20005594 TI - Prevalence of non-diabetic renal disease in patients with type 2 diabetes. AB - It is important to differentiate proteinuria from non-diabetic renal diseases (NDRD) or diabetic nephropathy in diabetic patients. The purpose of our study was to evaluate the prevalence of NDRD. A retrospective analysis was performed on diabetic patients who had undergone renal biopsy during a 6-year period. Our study revealed a high prevalence of NDRD in the diabetic population. Sixty-nine patients were investigated, 52.2% were diagnosed as NDRD and 47.8% as DN. Focal segmental glomerulosclerosis was the most common lesion found in patients with NDRD. We found a relationship between DN and fasting blood glucose level, systolic blood pressure, diastolic blood pressure, LVMI, intima-media thickening (IMT), and the presence of carotid plaques. Patients with NDRD had a lower incidence of diabetic retinopathy (DR). The absence of DR to differentiate NDRD had a sensitivity of 72.7%, a specificity 91.7%, and an ROC=0.822. Fasting blood glucose level had a sensitivity and specificity of 93.9% and 75%, respectively. Similarly, the use of IMT had sensitivity and specificity of 90% and 75.8%, respectively. In this study, we determined that the absence of DR, a lower fasting blood glucose level, and IMT is useful in differentiating NDRD from DN in diabetic patients with overt proteinuria. PMID- 20005595 TI - Evidence of non-functional redundancy between two pea h-type thioredoxins by specificity and stability studies. AB - The largest group of plant thioredoxins (TRXs) consists of the so-called h-type; their great number raises questions about their specific or redundant roles in plant cells. Pisum sativum thioredoxin h1 (PsTRXh1) and Pisum sativum thioredoxin h2 (PsTRXh2) are both h-type TRXs from pea (Pisum sativum) previously identified and biochemically characterized. While both are involved in redox regulation and show a high-sequence identity (60%), they display different behavior during in vitro and in vivo assays. In this work, we show that these two proteins display different specificity in the capturing of protein targets in vitro, by the use of a new stringent method. PsTRXh2 interacted with classical antioxidant proteins, whereas PsTRXh1 showed a completely different pattern of targeted proteins, and was able to capture a transcription factor. We also showed that the two proteins display very different thermal and chemical stabilities. We suggest that the differences in thermal and chemical stability point to a distinct and characteristic pattern of protein specificity. PMID- 20005596 TI - Stress-induced co-expression of two alternative oxidase (VuAox1 and 2b) genes in Vigna unguiculata. AB - Cowpea (Vigna unguiculata) alternative oxidase is encoded by a small multigene family (Aox1, 2a and 2b) that is orthologous to the soybean Aox family. Like most of the identified Aox genes in plants, VuAox1 and VuAox2 consist of 4 exons interrupted by 3 introns. Alignment of the orthologous Aox genes revealed high identity of exons and intron variability, which is more prevalent in Aox1. In order to determine Aox gene expression in V. unguiculata, a steady-state analysis of transcripts involved in seed development (flowers, pods and dry seeds) and germination (soaked seeds) was performed and systemic co-expression of VuAox1 and VuAox2b was observed during germination. The analysis of Aox transcripts in leaves from seedlings under different stress conditions (cold, PEG, salicylate and H2O2 revealed stress-induced co-expression of both VuAox genes. Transcripts of VuAox2a and 2b were detected in all control seedlings, which was not the case for VuAox1 mRNA. Estimation of the primary transcript lengths of V. unguiculata and soybean Aox genes showed an intron length reduction for VuAox1 and 2b, suggesting that the two genes have converged in transcribed sequence length. Indeed, a bioinformatics analysis of VuAox1 and 2b promoters revealed a conserved region related to a cis-element that is responsive to oxidative stress. Taken together, the data provide evidence for co-expression of Aox1 and Aox2b in response to stress and also during the early phase of seed germination. The dual nature of VuAox2b expression (constitutive and induced) suggests that the constitutive Aox2b gene of V. unguiculata has acquired inducible regulatory elements. PMID- 20005597 TI - Impairment of experience-dependent cortical plasticity in aged mice. AB - This study addresses the relationship between aging and experience-dependent plasticity in the mouse somatosensory cortex. Plasticity in the cortical representation of vibrissae (whiskers) was investigated in young (3 months), mature (14 months) and old (2 years) mice using [14C]2-deoxyglucose (2-DG) autoradiography. Plastic changes were evoked using two experimental paradigms. The deprivation-based protocol included unilateral deprivation of all but one row of whiskers for a week. In the conditioning protocol the animals were subjected to classical conditioning, where tactile stimulation of one row of whiskers was paired with an aversive stimulus. Both procedures evoked functional plasticity in the young group, expressed as a widening of the functional cortical representation of the spared or conditioned row. Aging had a differential effect on these two forms of plasticity. Conditioning-related plasticity was more vulnerable to aging: the plastic change was not detectable in mature animals, even though they acquired the behavioral response. Deprivation-induced plasticity also declined with age, but some effects were persistent in the oldest animals. PMID- 20005598 TI - Behavioral changes in aging female C57BL/6 mice. AB - Using a range of tests we have studied alterations in behavior with advancing age in female C57BL/6 (of Jackson origin), the golden standard on which most genetically engineered mice are back-crossed. In parallel, growth and survival data were collected. In a protected environment the 90% and 75% cohort survival age was 20 and 25 months, respectively, and the 50% cohort survival was 32 months. In mice, body weight increases continuously until 15-20 months of age, while in advanced age whole body weight drops. The body mass loss in senescence is associated with emergence of other aged phenotype features such as kyphosis, balding and loss of fur-color. Our behavioral data show that aging modulates certain aspects of basic behavior in a continuous manner, like explorative and locomotor activities. Advanced age associates with an acceleration of behavioral impairments evident in most of the tests used, including motor skill acquisition and memory consolidation. However, certain domains of mouse behavior were well preserved also in advanced age such as thermal noxious threshold and working memory as assessed by an object recognition task. The decreased drive to explore is suggested to be a key factor underlying many aspects of reduced performance including cognitive capacity during aging. Behavioral aging affects genetically closely related individuals housed under strictly standardized conditions differentially (Collier, T.J., Coleman, P.D., 1991. Divergence of biological and chronological aging: evidence from rodent studies. Neurobiol. Aging, 12, 685-693; Ingram, D.K., 1988. Motor performance variability during aging in rodents. Assessment of reliability and validity of individual differences. Ann. N.Y. Acad. Sci., 515, 70-96). Consistent with this a subpopulation of the 28-month-old mice showed an explorative activity similar to young-adult mice and a significantly stronger preference for a novel object than aged mice with a less explorative behavior. Thus, subtle environmental factors and epigenetic modifications may be important modulators of aging. PMID- 20005599 TI - Brain ERP components predict which individuals progress to Alzheimer's disease and which do not. AB - Predicting which individuals will progress to Alzheimer's disease (AD) is important in both clinical and research settings. We used brain Event-Related Potentials (ERPs) obtained in a perceptual/cognitive paradigm with various processing demands to predict which individual Mild Cognitive Impairment (MCI) subjects will develop AD versus which will not. ERP components, including P3, memory "storage" component, and other earlier and later components, were identified and measured by Principal Components Analysis. When measured for particular task conditions, a weighted set of eight ERP component_conditions performed well in discriminant analysis at predicting later AD progression with good accuracy, sensitivity, and specificity. The predictions for most individuals (79%) had high posterior probabilities and were accurate (88%). This method, supported by a cross-validation where the prediction accuracy was 70-78%, features the posterior probability for each individual as a method of determining the likelihood of progression to AD. Empirically obtained prediction accuracies rose to 94% when the computed posterior probabilities for individuals were 0.90 or higher (which was found for 40% of our MCI sample). PMID- 20005601 TI - Acceleration of cortical thinning in familial Alzheimer's disease. AB - BACKGROUND: MRI in presymptomatic autosomal dominant Alzheimer's disease mutation carriers (MC) provides an opportunity to detect changes that pre-date symptoms or clinical diagnosis. We used automated cortical thickness (CTh) measurement to compare the grey matter of such a group with cognitively normal controls. METHODS: 9 presymptomatic mutation carriers (4 PSEN1, 5 APP) and 25 healthy, age and sex-matched controls underwent longitudinal volumetric MRI brain imaging. CTh measurement was performed across the whole brain using a validated, automated technique. Four regions of interest (ROI) (entorhinal cortex (ERC), parahippocampal gyrus (PHG), posterior cingulate cortex and precuneus) and two control regions (paracentral and pericalcarine) were selected on the basis of imaging data in existing Alzheimer's disease (AD) literature. Linear mixed models were used to describe normal ageing in controls and the extent to which mean CTh in cases differed from controls according to time since clinical diagnosis, adjusting for normal ageing. RESULTS: An accelerating decline in CTh was observed across all ROI in the MC group. No such decline was demonstrated in the control regions for the MC group. Relative to controls, and adjusting for normal ageing, there was evidence (p=0.05, one-sided test) of lower CTh in the posterior cingulate up to 1.8 years prior to diagnosis and in the precuneus up to 4.1 years prior to diagnosis in the MC group. DISCUSSION: Automated CTh analysis is a relatively practical, rapid and effective technique for assessing subtle structural change in AD. There is evidence that cortical thickness is reduced in mutation carriers a number of years prior to clinical diagnosis. PMID- 20005600 TI - MRI-based volumetric measurement of the substantia innominata in amnestic MCI and mild AD. AB - The substantia innominata (SI) contains the nucleus basalis of Meynert, which provides the major cholinergic innervation to the entire cortical mantel and the amygdala; degeneration of nucleus basalis neurons correlates with cognitive decline in Alzheimer's disease (AD). However, whether SI atrophy occurs in individuals with amnestic mild cognitive impairment (aMCI) has not been examined thoroughly in vivo. In the present study, we developed a new protocol to measure volumetric changes in the SI from magnetic resonance imaging (MRI) scans. Participants consisted of 27 elderly controls with no cognitive impairment (NCI); 33 individuals with aMCI; and 19 patients with mild AD. SI volumes were traced on three consecutive gapless 1mm thick coronal slices. Results showed that SI volume was significantly reduced in the mild AD group compared to both NCI and aMCI participants; however, the NCI and aMCI groups did not differ from each other. Furthermore, a decrease in SI volume was related to impaired performance on declarative memory tasks even when attention was controlled. PMID- 20005602 TI - [Acute on chronic liver failure]. AB - Acute-on-Chronic Liver Failure (ACLF) is a recently introduced term defined as severe acute deterioration of an established liver disease. This entity usually develops after an acute insult. The main clinical manifestations are hepatorenal syndrome, hepatic encephalopathy and organ failure, with a high risk of death in the short term. The true incidence of ACLF remains difficult to determine due to confusions surrounding the definition of this entity, but seems to be 40% at 5 years in patients with advanced cirrhosis, which translates into 4,000 cases in Europe within this time span. The treatment of choice is liver transplantation. However, due to the shortage of suitable organs and morbidity and mortality in these patients, other options must be used. PMID- 20005603 TI - [Factors related to bullying in adolescents in Barcelona (Spain)]. AB - OBJECTIVES: Violence and bullying in the school setting are a serious social and health problem that have created great alarm in the last few years. We aimed to describe bullying and factors related to this phenomenon in students in the city of Barcelona. METHODS: We performed a cross-sectional study of a representative sample of 2,727 students from 66 secondary schools in Barcelona. Bullying was defined as having been laughed at, hit or marginalized four or more times, or as having been the target of all three of these behaviors at least once, in the last 12 months. To analyze the associations among bullying and related factors including sociodemographic variables, attitudes and behaviors, we used bivariate and multivariate logistic regression. RESULTS: The prevalence of bullying was 18.2%, 10.9% and 4.3% in boys and 14.4%, 8.5% and 4.5% in girls in the 8th, 10th and 12th grades, respectively. Factors associated with an increased likelihood of being bullied were negative mood states and violent behavior, while being older, alcohol consumption, cannabis use and going to bars and discos were negatively associated with being bullied. CONCLUSIONS: This study confirms that bullying is a serious problem in our context and identifies certain factors, such as negative mood states and other risk behaviors. These factors should be further analyzed in longitudinal studies in order to design and implement appropriate prevention programs. PMID- 20005604 TI - [Health and its determinants in the immigrant population of the region of Madrid]. AB - OBJECTIVES: To describe health status and its determinants in immigrants living in the region of Madrid and to compare these factors with those in the native born population. MATERIAL AND METHODS: We performed a descriptive, cross sectional study based on data from the Madrid Regional Health Survey 2007 (n=12,190). Subjects were classified as native-born or immigrants born in medium to-low-income countries and with less or more than 5 years of residence in Spain. The results were adjusted for age. Proportions and means were compared through the ji square test and linear regression. RESULTS: Compared with native-born residents, immigrants were younger, carried out mainly precarious manual jobs that did not match their educational level and had a lower income and less space in their dwellings. Both immigrant men and women smoked and drank less, but women showed a higher prevalence of overweight (10% more) and physical inactivity. Adherence to smear test and blood pressure screening was up to a 16% lower. Attendance at primary care and emergency facilities was less frequent in men and women with less residence time than in those with longer residence in Spain. Self perceived health and mental health were worse in women with 5 or more years of residence. Chronic diseases were similar in both populations but were less prevalent in immigrants. CONCLUSIONS: Immigrants' health status is not as unfavorable as could be expected from the higher prevalence of harmful determinants of health. PMID- 20005605 TI - [Reproductive and perinatal health indicators in immigrant and Spanish-born women in Catalonia and Valencia (2005-2006)]. AB - OBJECTIVES: To determine the prevalence of teenage maternity, preterm birth and low birth weight in Spanish and immigrant mothers from Latin America, eastern Europe, Maghreb and sub-Saharan Africa resident in Catalonia and Valencia from 2005 and 2006. METHODS: Using data from congenital metabolic disorders registers in both regions, proportions and 95% confidence intervals were obtained for the following: 1) mothers aged less than 20 years; 2) preterm (<37 weeks) and very preterm (<32 weeks) births; and 3) low birth weight (<2500g) and very low birth weight (<1500g) neonates. The calculations were performed for mothers from each of the geographical areas of origin (Spain, Latin America, Eastern Europe, Maghreb and Sub-Sahara). These proportions were compared in Spanish-born and immigrant women and the significance of differences was assessed using chi squared tests. RESULTS: The prevalence of teenage mothers was between three and five times higher in immigrants than in Spanish women, the highest rate being found in women from eastern Europe. Preterm births, very preterm births and very low birth weight were more frequent in eastern European women than in Spanish women. The prevalence of prematurity and very low birth weight was higher in sub Saharan mothers than in Spanish women. CONCLUSIONS: The number of births in teenage mothers was higher in immigrant mothers from all origins than in Spanish women. The highest rates of low birth weight and preterm births were found in women from eastern Europe and sub-Saharan Africa. PMID- 20005607 TI - Surfing or still drowning? Student nurses' Internet skills. AB - A study into student nurses' ability to use the Internet was published in Nurse Education Today in 2004. This paper repeats the research with a cohort of students starting their pre-registration programme in a UK university in 2007. In 2004 students were reported as having poor Internet skills, and as not being frequent users of the Internet. In this study students were found to have significantly better ability to carry out basic tasks and significantly higher levels of Internet use. Their ability to apply these skills to more complex information literacy tasks however had not increased, with more than half of all students saying they found far too much irrelevant information when searching for specific information on the Internet. The earlier study found that skills and age were not related, which appears to still be the case. The need for these skills is increasing as education, lifelong learning, and patient information are all increasingly drawing on the developing Internet. Nurse education however is not integrating the skill and knowledge base essential to support this into pre registration programmes, and the evidence suggests that this will not happen without active management. PMID- 20005606 TI - Future challenges for nursing education--a European perspective. AB - In Europe, there have been reforms in nursing education during last years and many political papers have been published. The reforms have given need for harmonising nursing education. In spite of that, there are differences in nursing education system in Europe. In this paper, we describe some main policy papers in the field of nursing education and identify selected future challenges. These challenges have been named for developing cross-cultural collaboration, clinical learning environment, role of patients and teacher education. PMID- 20005608 TI - Case seminars open doors to deeper understanding - Nursing students' experiences of learning. AB - BACKGROUND: The Case Method is a teaching method in which cases from real life inspire students to actively seek knowledge that they discuss in structured seminars. Case seminars in health education have been evaluated, compared and discussed, but descriptions that can help us understand how students learn in the seminars have not previously been published. In a Swedish nursing programme, where case seminars have been used for several years, students were asked to write about their experiences of learning in the seminars. The aim of the present study was to describe this learning process from the students' point of view. METHOD: Written data were analysed using content analysis. FINDINGS: A theme concerning how the Case Method opens doors to deeper understanding was identified as a thread running through different codes and categories. Students described the importance of new perspectives and their wish to participate in discussions with other students. The students indicated that the structure, which involved pre-prepared cases and writing on the white board, positioned their own knowledge in a wider context and that the learning atmosphere enabled everyone to participate. CONCLUSIONS: The Case Method seems to involve students in a way that deepens their understanding and critical thinking. PMID- 20005609 TI - How to be a professor: What Taylor et al. didn't tell you! PMID- 20005610 TI - Legality, the Web and nurse educators. AB - Nurse lecturers are becoming more aware of the opportunities that web 2.0 offers and are slowly moving into the world of cyber-teaching. However, few consider the legal ramifications of this new teaching trend and how vulnerable they may be when posting information or teaching materials to the World Wide Web. The purpose of this paper is to explore the legal issues inherent in the use of virtual learning technologies. This paper poses a number of questions that novices in the world of virtual learning technologies may wish to consider. The paper makes some recommendation which may help to ensure that not only can nurse educators fully exploit the opportunities offered by the World Wide Web but they can do so in a legally responsible manner. PMID- 20005611 TI - Assessment of the radiological impact of oil refining industry. AB - The field of radiation protection and corresponding national and international regulations has evolved to ensure safety in the use of radioactive materials. Oil and gas production processing operations have been known to cause naturally occurring radioactive materials (NORMs) to accumulate at elevated concentrations as by-product waste streams. A comprehensive radiological study on the oil refining industry in Egypt was carried out to assess the radiological impact of this industry on the workers. Scales, sludge, water and crude oil samples were collected at each stage of the refining process. The activity concentration of (226)Ra, (232)Th and (40)K were determined using high-resolution gamma spectrometry. The average activity concentrations of the determined isotopes are lower than the IAEA exempt activity levels for NORM isotopes. Different exposure scenarios were studied. The average annual effective dose for workers due to direct exposure to gamma radiation and dust inhalation found to be 0.6 microSv and 3.2 mSv, respectively. Based on the ALARA principle, the results indicate that special care must be taken during cleaning operations in order to reduce the personnel's exposure due to maintenance as well as to avoid contamination of the environment. PMID- 20005612 TI - Escaping radioactivity from coal-fired power plants (CPPs) due to coal burning and the associated hazards: a review. AB - Coal, like most materials found in nature, contains trace quantities of the naturally occurring primordial radionuclides, i.e. of (40)K and of (238)U, (232)Th and their decay products. Therefore, the combustion of coal results in the released into the environment of some natural radioactivity (1.48 TBq y(-1)), the major part of which (99%) escapes as very fine particles, while the rest in fly ash. The activity concentrations of natural radionuclides measured in coals originated from coal mines in Greece varied from 117 to 435 Bq kg(-1) for (238)U, from 44 to 255 Bq kg(-1) for (226)Ra, from 59 to 205 Bq kg(-1) for (210)Pb, from 9 to 41 Bq kg(-1) for (228)Ra ((232)Th) and from 59 to 227 Bq kg(-1) for (40)K. Fly ash escapes from the stacks of coal-fired power plants in a percentage of 3 1% of the total fly ash, in the better case. The natural radionuclide concentrations measured in fly ash produced and retained or escaped from coal fired power plants in Greece varied from 263 to 950 Bq kg(-1) for (238)U, from 142 to 605 Bq kg(-1) for (226)Ra, from 133 to 428 Bq kg(-1) for (210)Pb, from 27 to 68 Bq kg(-1) for (228)Ra ((232)Th) and from 204 to 382 Bq kg(-1) for (40)K. About 5% of the total ash produced in the coal-fired power plants is used as substitute of cement in concrete for the construction of dwellings, and may affect indoor radiation doses from external irradiation and the inhalation of radon decay products (internal irradiation) is the most significant. The resulting normalized collective effective doses were 6 and 0.5man-Sv(GWa)(-1) for typical old and modern coal-fired power plants, respectively. PMID- 20005613 TI - Alterations in three-dimensional joint kinematics of anterior cruciate ligament deficient and -reconstructed knees during walking. AB - BACKGROUND: High risk of cartilage degeneration and premature osteoarthritis development has been clinically observed in anterior cruciate ligament (ACL) deficient knees. The risk has not been significantly reduced even after ACL reconstructive surgery. It was hypothesized that three-dimensional knee joint kinematics has been altered after ACL injury, and the biomechanical alteration has not been fully restored to a normal level after reconstructive surgery. METHODS: Spatiotemporal parameters and three-dimensional knee joint rotations and translations were measured in ACL-deficient, ACL-reconstructed, and ACL-intact knees during level walking. The variables were compared between the ACL-deficient and ACL-intact knees, as well as between the ACL-reconstructed and ACL-intact knees. FINDINGS: Altered spatiotemporal variables and key event timings in a gait cycle were observed in both ACL-deficient and ACL-reconstructed subjects. Significant reduction of extension was observed in the ACL-deficient knees during midstance and in the ACL-reconstructed knees during swing phase. Greater varus and internal tibial rotation were identified in the ACL-deficient knees. Although being small in magnitude, these secondary kinematic alterations were consistent throughout the whole gait cycle, and such trends were not eliminated in the ACL reconstructed knees. INTERPRETATION: Significant abnormalities of spatiotemporal performance and three-dimensional joint kinematics during walking were identified in the ACL-deficient knees. The ACL-reconstructed knees exhibited some improvement in joint kinematics, but not being fully restored to a normal level. Identification of biomechanical alterations during daily activities in ACL deficient and ACL-reconstructed knees could help better understand clinical outcomes and seek improvement in surgical technique and rehabilitation regimen for ACL injury treatment. PMID- 20005614 TI - Wavelet analysis based on time-frequency information discriminate chronic ankle instability. AB - BACKGROUND: Ankle sprains are one of the most common lower extremity injuries. Real time human motion tracking is an accurate, inexpensive and portable system to obtain kinematic and kinetic measurements. The purpose of this study was to discriminate between subjects with chronic ankle instability and subjects with stable ankles through inertial tracking technology and force plates. METHODS: Twelve subjects (mean (SD) 23.16 (5.32) years, 174.83 (8.78) cm, 73.58 (17.10) kg) with stable ankles and 13 (mean (SD) 24.69 (5.91) years, 173.31 (9.07) cm, 69.61 (15.32) kg) with chronic ankle instability performed the Star Excursion Balance Test. Time-frequency information based on wavelet decomposition was used for analysing all signals. FINDINGS: Dynamic balance impairment associated with chronic ankle instability was observed in the peak amplitude in the wavelet approximation as well as the absolute sum of the coefficients of the wavelet details of the acceleration, orientation and force signals. These results were found despite Star Excursion Balance Test performance during anterior, posteromedial and posterolateral excursions lead to similar specific reach distances in both limbs in either the chronic ankle instability or stable ankle groups. INTERPRETATION: These parameters could be of great interest in detecting dynamic balance impairment in individuals at risk of sprains that might otherwise go undetected by only reach distance assessment. PMID- 20005615 TI - Optimising chronic myeloid leukaemia therapy in the face of resistance to tyrosine kinase inhibitors--a synthesis of clinical and laboratory data. AB - The introduction of imatinib, a tyrosine kinase inhibitor (TKI) that targets the BCR-ABL protein, has revolutionised the treatment of chronic myeloid leukaemia (CML), producing high rates of response that have been durable in many patients. However, because of intrinsic or acquired mechanisms of imatinib resistance, in addition to the persistence of leukaemic stem cells that are resistant to imatinib-induced apoptosis, imatinib treatment does not appear to be curative. Cytogenetic and molecular monitoring enable the identification of patients showing signs of treatment failure and can be used to guide choices regarding subsequent therapeutic options, including imatinib dose escalation, treatment with a secondary TKI or, in selected cases, allogeneic stem cell transplant (allo SCT). Although these alternative therapies may overcome imatinib resistance, long term remission or cure from CML is likely to require development of novel interventions that effectively eliminate CML stem cells (Ph+HSC). PMID- 20005617 TI - Why do interventions work in some places and not others: a breastfeeding support group trial. AB - In a cluster randomised controlled trial of a policy to provide community breastfeeding support groups in Scotland, breastfeeding rates declined in 3 of 7 intervention localities. From a preliminary study, we expected breastfeeding outcomes to vary and we prospectively used qualitative and quantitative methods to ask why. Ethnographic in-depth interviews, focus groups, observations and survey data were analysed to build seven embedded case studies. A pyramidal model of how primary health service organisations implemented the policy was constructed prior to knowing trial outcomes to minimise bias. Informed by a realist approach, the model explained variation in (a) policy implementation (b) the breastfeeding outcomes, whereas the quantity of intervention delivered did not. In the three localities where breastfeeding rates declined, negative aspects of place including deprivation, unsuitable premises and geographical barriers to inter-professional communication; personnel resources including staff shortages, high workload and low morale; and organisational change predominated (the base model tiers). Managers focused on solving these problems rather than delivering the policy and evidence of progress to the higher model tiers was weak. In contrast, where breastfeeding rates increased the base tiers of the model were less problematic, there was more evidence of leadership, focus on the policy, multi-disciplinary partnership working and reflective action cycles (the higher model tiers). We advocate an ethnographic approach to the design and evaluation of complex intervention trials and illustrate how this can assist in developing an explanatory model. More attention should be given to the complex systems within which policies and interventions occur, to identify and understand the favourable conditions necessary for a successful intervention. PMID- 20005618 TI - Interpersonal perception in the context of doctor-patient relationships: a dyadic analysis of doctor-patient communication. AB - Doctor-patient communication is an interpersonal process and essential to relationship-centered care. However, in many studies, doctors and patients are studied as if living in separate worlds. This study assessed whether: 1) doctors' perception of their communication skills is congruent with their patients' perception; and 2) patients of a specific doctor agree with each other about their doctor's communication skills. A cross-sectional study was conducted in three provinces in Canada with 91 doctors and their 1749 patients. Doctors and patients independently completed questions on the doctor's communication skills (content and process) after a consultation. Multilevel modeling provided an estimate of the patient and doctor variance components at both the dyad-level and the doctor-level. We computed correlations between patients' and doctors' perceptions at both levels to assess how congruent they were. Consensus among patients of a specific doctor was assessed using intraclass correlation coefficient (ICC). The mean score of the rating of doctor's skills according to patients was 4.58, and according to doctors was 4.37. The dyad-level variance for the patient was .38 and for the doctor was .06. The doctor-level variance for the patient ratings was .01 and for the doctor ratings, .18. The correlation between both the patients' and the doctors' skills' ratings scores at the dyad-level was weak. At the doctor-level, the correlation was not statistically significant. The ICC for patients' ratings was .03 and for the doctors' ratings .76. Overall, this study suggests that doctors and their patients have a very different perspective of the doctors' communication skills occurring during routine clinical encounters. PMID- 20005616 TI - Mechanisms of attentional biases towards threat in anxiety disorders: An integrative review. AB - A wealth of research demonstrates attentional biases toward threat in the anxiety disorders. Several models have been advanced to explain these biases in anxiety, yet the mechanisms comprising and mediating the biases remain unclear. In the present article, we review evidence regarding the mechanisms of attentional biases through careful examination of the components of attentional bias, the mechanisms underlying these components, and the stage of information processing during which the biases occur. Facilitated attention, difficulty in disengagement, and attentional avoidance comprise the components of attentional bias. A threat detection mechanism likely underlies facilitated attention, a process that may be neurally centered around the amygdala. Attentional control ability likely underlies difficulty in disengagement, emotion regulation goals likely underlie attentional avoidance, and both of these processes may be neurally centered around prefrontal cortex functioning. The threat detection mechanism may be a mostly automatic process, attentional avoidance may be a mostly strategic process, and difficulty in disengagement may be a mixture of automatic and strategic processing. Recommendations for future research are discussed. PMID- 20005619 TI - Mineralization of the stylohyoid ligament complex in a Jordanian sample: a clinicoradiographic study. AB - PURPOSE: To record the prevalence of anatomic variations and the associated symptoms of mineralized stylohyoid complex (MSHC) in a Jordanian sample, and to investigate the possible factors responsible for mineralization. PATIENTS AND METHODS: This study was conducted at the Dental Teaching Clinics of Jordan University of Science and Technology in patients referred for panoramic radiographs. The MSHC, on either side of panoramic radiographs, was mapped out on tracing paper, the length directly measured, and the true length calculated. The true length of the MSHC was used to classify every side into short, long, or elongated, and the radiographic type of long and elongated MSHCs was determined as elongated, pseudoarticulated, or segmented. RESULTS: A total of 867 MSHCs were traced with an overall average length of the 23.1 mm. The most frequent class and type were long (53.3%) and elongated (41.6%), respectively. There was a significantly (P < .05) progressive increase in the length of the MSHC. The frequency of both of elongated and pseudoarticulated types increased and that of the segmented type decreased with aging. The mean length of the MSHC for patients with a history of tonsillectomy (26.29 mm) was significantly higher (P < .05) than its value in patients without a history of tonsillectomy (22.70). There was also a significantly (P < .05) progressive increase in the length of the MSHC as the age when tonsillectomy was performed increased. The mean length of the MSHC for cases with history of recurrent tonsillitis (6.99 mm) was significantly higher (P < .05) than for those without (2.42 mm). The mean length of the MSHC for patients with a history of head-and-neck or whole-body trauma was not statistically significantly different (P > .05) from the value for the nontraumatized patients. The mean length of the MSHC for patients with arthritis (27.70 mm) was significantly higher (P < .05) than that in patients without arthritis (22.53 mm). The highest frequency of MSHC palpated in the tonsillar fossa was significantly (P < .05) related to the elongated class (44; 64.8%) and type (100; 52%). The mean length of the MSHC was significantly higher in patients who reported having 4 symptoms, but only 1 symptom had a significant association with the type of MSHC. CONCLUSION: History of recurrent tonsillitis has a major effect on the true length of the MSHC, and trauma is not a significant factor for the development of longer MSHC. PMID- 20005620 TI - Lower extremity compartment syndrome associated with hypotensive general anesthesia for orthognathic surgery: a case report and review of the disease. PMID- 20005621 TI - Extraneural soft tissue perineurioma of the oral mucosa. PMID- 20005622 TI - A rare case of IgG4-related sclerosing disease of the maxillary sinus associated with bone destruction. PMID- 20005623 TI - "Early rupture of membranes" after the spontaneous onset of labor as a risk factor for cesarean delivery. AB - OBJECTIVE: The objective of this study was to examine if patients with "early rupture of membranes (ROM)" after spontaneous onset of labor are at increased risk of cesarean section. STUDY DESIGN: The rate of cesarean section was examined in 447 term singleton nulliparas who were admitted after the spontaneous onset of labor. The cases were divided into 2 groups: (1) "early ROM", defined as ROM at a cervical dilatation <4 cm (n=109); and (2) "late ROM", ROM at a cervical dilatation >or=4 cm (n=338). RESULTS: (1) "Early ROM" occurred in 24.4% of the cases and the overall cesarean section rate was 5.6%; (2) there were no significant differences in the clinical characteristics including prepregnancy BMI, proportion of complicated pregnancies, total duration of labor, proportion of regional anesthesia, gestational age at delivery, and birthweight between the two groups of cases. However gravidas with "early ROM" were of advanced maternal age and had less cervical dilation on admission, shorter duration of 1st stage of labor, and more frequent use of oxytocin augmentation; (3) patients with "early ROM" had a threefold higher rate (11.9% vs. 3.6%) of cesarean section and a fourfold higher rate (11.9% vs. 3.0%) of cesarean section due to failure of progress than did those with "late ROM" (p<0.005 for each); (3) 92% (23/25) of cesarean sections were performed due to failure to progress; and (4) there was no significant difference in the rate of histologic chorioamnionitis between the two groups of cases. CONCLUSION: "Early ROM" after the spontaneous onset of labor is a risk factor for cesarean section in term singleton nulliparas. PMID- 20005624 TI - First manifestation of citrullinemia type I as differential diagnosis to postpartum psychosis in the puerperal period. PMID- 20005625 TI - Roles of abiotic losses, microbes, plant roots, and root exudates on phytoremediation of PAHs in a barren soil. AB - Phytoremediation is an emerging technology for the remediation of polycyclic aromatic hydrocarbons (PAHs). In this study, pot experiments were conducted to evaluate the efficacy of phytoremediation of phenanthrene and pyrene in a typical low organic matter soil (3.75 g kg(-1)), and the contribution proportions of abiotic losses, microbes, plant roots, and root exudates were ascertained during the PAHs dissipation. The results indicated that contribution of abiotic losses from this soil was high both for phenanthrene (83.4%) and pyrene (57.2%). The contributions of root-exudates-enhanced biodegradation of phenanthrene (15.5%) and pyrene (21.3%) were higher than those of indigenous microbial degradation. The role of root exudates on dissipation of phenanthrene and pyrene was evident in this experiment. By the way, with the increasing of ring numbers in PAHs structures, the root-exudates-enhanced degradation became more and more important. BIOLOG-ECO plate analysis indicated that microbial community structure of the soil receiving root exudates had changed. The removal efficiency and substrate utilization rate in the treatment with plant roots were lower than the treatment only with root exudates, which suggested that possible competition between roots and microbes for nutrients had occurred in a low organic matter soil. PMID- 20005626 TI - The role of open and closed curing conditions on the leaching properties of fly ash-slag-based geopolymers. AB - This study deals with the synthesis of geopolymers from co-fired fly ash and blast furnace slags. Geopolymer bodies were simultaneously synthesized in open and closed curing conditions in order to elucidate the role of this parameter on their resultant properties. Open curing conditions produce solid bodies characterized by high porosity, low compressive strength and exacerbated leaching of certain oxyanionic metalloids. By contrast, protected curing promotes the binder development, giving rise to higher strength and less porous systems. This imposes physical restrictions to leaching which decreases and/or retards releases of oxyanionic metalloids in comparison to open curing conditions. Fly ash-slag based geopolymers may immobilize a number of trace pollutants such as Be, Bi, Cd, Co, Cr, Cu, Nb, Ni, Pb, REE, Sn, Th, U, Y and Zr, regardless of the curing conditions. Due to geopolymers displaying weak assimilation capacity for oxyanionic species, their successful regarding oxyanionic retention is strongly dependent on porosity and therefore on curing conditions applied. PMID- 20005627 TI - Surface water monitoring in the mercury mining district of Asturias (Spain). AB - Systematic monitoring of surface waters in the area of abandoned mine sites constitutes an essential step in the characterisation of pollution from historic mine sites. The analytical data collected throughout a hydrologic period can be used for hydrological modelling and also to select appropriate preventive and/or corrective measures in order to avoid pollution of watercourses. Caudal River drains the main abandoned Hg mine sites (located in Mieres and Pola de Lena districts) in Central Asturias (NW Spain). This paper describes a systematic monitoring of physical and chemical parameters in eighteen selected sampling points within the Caudal River catchment. At each sampling station, water flow, pH, specific conductance, dissolved oxygen, salinity, temperature, redox potential and turbidity were controlled "in situ" and major and trace elements were analysed in the laboratory. In the Hg-mineralised areas, As is present in the form of As-rich pyrite, realgar and occasionally arsenopyrite. Mine drainage and leachates from spoil heaps exhibit in some cases acidic conditions and high As contents, and they are incorporated to Caudal River tributaries. Multivariate statistical analysis aids to the interpretation of the spatial and temporary variations found in the sampled areas, as part of a methodology applicable to different environmental and geological studies. PMID- 20005628 TI - Enzymatic bioremediation of cashew nut shell liquid contamination. AB - Cashew nut shell liquid (CNSL), a by-product of the cashew kernel industry, is a caustic, viscous, dark liquid. The process is done manually, which leaves stains on the hands of the workers. The aim was to find the utility of enzymes, oxidoreductases and proteases for the bioremediation of CNSL, which contains phenolics, mainly cardanol (60-65%). The results show that peroxidase reduced the color of the CNSL solution by polymerization and precipitation, where as laccase, papain and fungal and bacterial protease degraded the phenolic constituents. The degradation was mainly at the double bonds of the C15 hydrocarbon chain of the cardanol. To improve the enzyme stability, laccase and papain was separately immobilized in alginate-starch beads. Immobilized laccase can degrade 28.6% CNSL within 2 h, where as papain takes longer duration, and at 73 h, the adsorbed phenols on the alginate (45.86%) also got degraded. MALDI-TOF MS revealed that, immobilized laccase-papain beads combination; 1:1 (w/w) degraded 60% of the cardanol and some phenolic compounds having molecular mass of 374, 390 and 407. These beads are active and stable in aqueous media, can be used to prepare a mild, nontoxic, ecofriendly, cost effective hand wash solution for the removal of phenolic stains. PMID- 20005629 TI - A comparative study of electrocoagulation and coagulation of aqueous suspensions of kaolinite powders. AB - Removal of kaolinite particles from their synthetically prepared suspensions was studied by electrocoagulation and coagulation to investigate the effect of such operating parameters as initial pH, coagulant dosage, applied voltage, current density, and time. Coagulation was more effective in a wider pH range (pH 5-8) than electrocoagulation which yielded optimum effectiveness in a relatively narrower pH range around 9, where, in both methods, these pH values corresponded to near-zero zeta potentials of kaolinite particles. The mechanism for both coagulation methods was aggregation through charge neutralization and/or enmeshment in aluminum hydroxide precipitates. The kinetics of electrocoagulation was very fast (<10 min) in approaching a residual turbidity, which could be modeled with a second-order rate equation. PMID- 20005630 TI - Characterization and mechanistic analysis of the visible light response of cerium and nitrogen co-doped TiO2 nano-photocatalyst synthesized using a one-step technique. AB - Cerium and nitrogen co-doped anatase TiO(2) nanoparticles were synthesized using a one-step technique via a modified sol-gel process and characterized by XRD, BET, DRS, Raman and XPS. The photocatalytic mechanism of the degradation of methylene blue (MB) under fluorescent light and visible light irradiation was studied. Co-doping cerium and nitrogen in the crystal lattice of TiO(2) narrowed the band gap from 2.40 eV (Ce-doped TiO(2)) to 2.21 eV (Ce/N co-doped TiO(2)). Ce(4+)/Ce(3+) pairs, oxynitride species and Ti-O-N and Ti-O-Ce bonds were determined by XPS. The recombination of photogenerated electron-hole pairs was inhibited due to the synergistic effect of doping with Ce(4+)/Ce(3+) ions and N atoms. The optimal doping ratio was 0.70% Ce and 0.70% N using MB photocatalytic degradation under fluorescent light and visible light irradiation (lambda>420 nm). The enhanced photocatalytic degradation under visible light irradiation was attributed to the increasing number of photogenerated OH radicals. The recombination of photogenerated e(-)-h(+) was attributed to be the key factor for the decrease in the photocatalytic degradation efficiency of MB. PMID- 20005632 TI - Parent attitudes towards medical student attendance and interaction in the paediatric burns outpatient clinic. AB - Medical student attendance in the Outpatient Department is not only essential for the teaching of paediatric burns, but is also important in the recruitment of promising individuals for the growth of our speciality. In 2008, 110 consecutive parents were asked to complete written surveys before the start of their clinic appointment. Only one parent per family could choose to complete the survey. The response rate was 100%. Parents from a diverse range of cultural, educational and socioeconomic were represented in this study. Eighteen parents (16.4%), 48 parents (46.3%) and 44 parents (40.0%) considered their child's burn to be severe, moderate and mild, respectively. One hundred and nine parents (99.1%) accepted the attendance of medical students. Forty-two parents (38.5%) preferred fewer than 3 students, 35 parents (32.1%) would be comfortable with 3-5 medical students and 32 parents (29.4%) could accept more than 6 medical students. One hundred and two parents (92.7%) would allow students to physically interact with their children and 108 parents (99.2%) would allow medical students to freely ask questions in burns clinic. Using logistic regression analysis, we found that high income earners would be comfortable with fewer medical students attending clinic than low income earners (p=0.007). Also, younger parents (p=0.002) and parents from families who made less than A$25,000 a year (p=0.009), believed that they could perceive 'a lot of benefit' from observing medical students being taught, whereas older parents and parents from higher income families responded more often with a perception of only 'some benefit'. This first study in a paediatric outpatient setting shows that parents are overwhelmingly prepared to have medical students involved in the care of their child. PMID- 20005633 TI - Also about forgotten staples. PMID- 20005634 TI - Estrogen regulates cytoskeletal flexibility, cellular metabolism and synaptic proteins: A proteomic study. AB - Estrogen (E2) influences brain function to induce gender differences in neuronal processes. In contrast to its well-described effects on signaling systems and gene transcription factors, our knowledge of E2-regulated protein networks is rather limited. Thus, we examined changes in protein expression patterns in the whole brains of ovariectomized mice after 24h estrogen exposure using two dimensional differential gel electrophoresis. Interpretation of our network-based hypothesis suggested that E2 regulates synaptic proteins and processes, increases cytoskeletal flexibility and alters glucose consumption in the brain. We verified the predicted reduced basal synaptic activity using in vivo microdialysis in conscious mice, showing that E2 decreases the extracellular concentrations of certain amino acids in two different brain areas (in the striatum and in the hypothalamus) and that this is independent from the E2 receptor densities. Our data reveal that E2 induces minor, but substantial changes to functionally different protein networks at the whole brain level, and as a cumulative effect, it adjusts the brain steady-state condition to a more flexible state. PMID- 20005635 TI - Are cancer stem cells concentrated in more alkaline hypoxic regions of tumors? AB - We wonder if the most viable hypoxic cancer stem cells concentrate in more alkaline regions of tumors, favoring their survival and evolution. Alternately, or in addition, do some cancer stem cells themselves maintain a more alkaline internal environment, achieving the same result. Based upon the response of cultured cells, including stem cells, to a certain degree of hypoxia and of most if not all proliferating cells to a somewhat more alkaline ambient and especially endogenous pH, their survival and proliferation should be favored. The broad outline of the argument, abstracted from a number of the available examples is developed: that the survival of cancer stem cells is favored by these conditions, contributing to their limited response to various therapies and their subsequent development of more malignant properties. PMID- 20005636 TI - On the segmental and tagmatic phenomena inferred from referred pain and autonomic concomitants. AB - Visceral injury induces referred pain, autonomic concomitants, skeletal muscle spasm, and hyperalgesia of segmental pattern on the specific area of somatic tissue. Therefore, since visceral referred pain does not mean only a feeling which is considered as misperception of brain but accompanies accessory phenomena, referred phenomenon can be more reasonable expression. Visceral referred phenomenon has segmental pattern and is shown on the segmental constituents related to viscus and further spreads to multi-segments through central sensitization if visceral injury become severe and chronic. Segmental pattern observed in our body is not a feature developing only in human. Segmentation of animals is a general concept in body organization from Annelida to Vertebrata. The evolutionary advent of segmentation made possible development of much greater complexity in structure and function. Segmentation increased efficiency of movements, and further made fine neural control of movements possible. It is definite that segmentation of body bestows obvious advantages to an animal. If abnormality in a segment develops, to lessen energy expenditure of an organism consisting of multi-segments, the organism sacrifices the abnormal segment for the whole segments via defunctionalization. Defunctionalization of abnormal segment is just segmental phenomenon. Functional unit of grouped segments called as tagma in Arthropoda or its equivalent in Vertebrata could also show this phenomenon if abnormality develops in tagma or its equivalent. Visceral referred pain, referred pain originating from musculoskeletal structure, complex regional pain syndrome I and II are representatives of segmental or tagmatic phenomena. PMID- 20005637 TI - Radiosurgery for craniopharyngioma. AB - PURPOSE: To analyze the outcomes of gamma knife stereotactic radiosurgery (SRS) for residual or recurrent craniopharyngiomas and evaluate the factors that optimized the tumor control rates. METHODS AND MATERIALS: A total of 46 patients with craniopharyngiomas underwent 51 SRS procedures at University of Pittsburgh between 1988 and 2007. The median tumor volume was 1.0 cm(3) (range, 0.07-8.0). The median prescription dose delivered to the tumor margin was 13.0 Gy (range, 9 20). The median maximal dose was 26.0 Gy (range, 20-50). The mean follow-up time was 62.2 months (range, 12-232). RESULTS: The overall survival rate after SRS was 97.1% at 5 years. The 3- and 5-year progression-free survival rates (solid tumor control) were both 91.6%. The overall local control rate (for both solid tumor and cyst control) was 91%, 81%, and 68% at 1, 3, and 5 years, respectively. No patients with normal pituitary function developed hypopopituitarism after SRS. Two patients developed homonymous hemianopsia owing to tumor progression after SRS. Among the factors examined, complete radiosurgical coverage was a significant favorable prognostic factor. CONCLUSION: SRS is a safe and effective minimally invasive option for the management of residual or recurrent craniopharyngiomas. Complete radiosurgical coverage of the tumor was associated with better tumor control. PMID- 20005638 TI - An effective preoperative three-dimensional radiotherapy target volume for extremity soft tissue sarcoma and the effect of margin width on local control. AB - PURPOSE: There is little information on the appropriate three-dimensional (3D) preoperative radiotherapy (XRT) volume for extremity soft-tissue sarcomas (STS). We retrospectively analyzed the pattern of local failure (LF) to help elucidate optimal field design. METHODS AND MATERIALS: We analyzed the 56 patients who underwent computed tomography-planned XRT for Stage I to III extremity STS between June 2000 and December 2006. Clinical target volume (CTV) included the T1 post-gadolinium-defined gross tumor volume with 1- to 1.5-cm radial and 3.5-cm longitudinal margins. Planning target volume expansion was 5 to 7 mm, and >or=95% of dose was delivered to the planning target volume. Preoperative XRT was 44 to 50.4 Gy (median, 50). Postoperative boost of 10 to 20 Gy was given to 12 patients (6 with positive and 6 with close margins). RESULTS: Follow-up ranged from 15 to 76 months (median, 41 months). The 5-year local control, freedom from distant metastasis, disease-free survival, and overall survival were 88.5%, 80.0%, 77.5% and 82.8%, respectively. Three patients (all with positive margin) experienced local failure (LF) as first relapse (2 isolated, 1 with distant failure), and 2 additional patients (all with margin<1 mm) had late LF after distant metastasis. The LFs were within the CTV in 3 patients and within and also extending beyond the CTV in 2 patients. CONCLUSIONS: These target volume definitions appear to be appropriate for most patients. No local recurrences were observed with surgical margins >or=1 mm, and it appears that these may be adequate for patients with extremity STS treated with preoperative radiotherapy. PMID- 20005639 TI - A comparison of dose-response models for the parotid gland in a large group of head-and-neck cancer patients. AB - PURPOSE: The dose-response relationship of the parotid gland has been described most frequently using the Lyman-Kutcher-Burman model. However, various other normal tissue complication probability (NTCP) models exist. We evaluated in a large group of patients the value of six NTCP models that describe the parotid gland dose response 1 year after radiotherapy. METHODS AND MATERIALS: A total of 347 patients with head-and-neck tumors were included in this prospective parotid gland dose-response study. The patients were treated with either conventional radiotherapy or intensity-modulated radiotherapy. Dose-volume histograms for the parotid glands were derived from three-dimensional dose calculations using computed tomography scans. Stimulated salivary flow rates were measured before and 1 year after radiotherapy. A threshold of 25% of the pretreatment flow rate was used to define a complication. The evaluated models included the Lyman Kutcher-Burman model, the mean dose model, the relative seriality model, the critical volume model, the parallel functional subunit model, and the dose threshold model. The goodness of fit (GOF) was determined by the deviance and a Monte Carlo hypothesis test. Ranking of the models was based on Akaike's information criterion (AIC). RESULTS: None of the models was rejected based on the evaluation of the GOF. The mean dose model was ranked as the best model based on the AIC. The TD(50) in these models was approximately 39 Gy. CONCLUSIONS: The mean dose model was preferred for describing the dose-response relationship of the parotid gland. PMID- 20005640 TI - [Can medical treatment be the first-line treatment for most ectopic pregnancies? A series of 202 patients]. AB - OBJECTIVES: To assess the efficacy and safety of a liberal policy of medical treatment for ectopic pregnancies (EUP). MATERIAL AND METHODS: Retrospective study of EUP that received active management between January 1999 and December 2003. Patients with rupture or near-rupture and those who refused medical treatment received surgical treatment. Other patients were treated by methotrexate. RESULTS: Two hundred and two EUP were managed; 26 % were treated surgically, 74 % medically. The success rate of medical treatment was 83 %. A ss hCG threshold of 2526 IU/l was selected. Under this level, the success rate was 90.7 % compared with 68 % when it was higher (p=0.001). Cardiac activity tripled the failure rate. Morbidity with medical treatment consisted of an augmentation in transaminases in 18.8 % and one case of severe dermatosis (0.7 %). Almost half of the patients treated medically required hospitalizations. CONCLUSION: The extension of medical treatment in our population to 74 % of all EUP yielded a high success rate, 83 %. PMID- 20005641 TI - [Stillbirth and tight umbilical cord knot: obviously guilty?]. AB - A healthy woman, gravida 1, para 1, spontaneously delivered a stillborn at 40 weeks of gestation, presenting with a tight umbilical cord knot. Fetopathological examination confirmed the main but not exclusive role of the knot in this intra uterine fetal death (IUFD). Knot of the umbilical cord is a rare occurrence, which is usually an incidental finding at delivery. However, it is associated with an increase in perinatal mortality. Only a complete fetopathological examination can state causality between knot and IUFD, exclude another fetal or placental cause of death and consequently reassure the parents for the prognosis of another pregnancy. PMID- 20005642 TI - Sex differences in the effects of allopregnanolone on yohimbine-induced reinstatement of cocaine seeking in rats. AB - Sex differences exist in several aspects of cocaine abuse, and recent research suggests that this may be due, in part, to differential sensitivity to stress. Women, compared to men, exhibit greater stress-induced cocaine craving and responses to both cocaine and stress fluctuate during phases of the hormonal cycle. The goal of the present study was to compare male and female rats on the maintenance and extinction of cocaine seeking and on an animal model of stress induced relapse by administering the pharmacological stressor yohimbine. An additional goal was to examine possible sex-specific treatment effects of the progesterone metabolite, allopregnanolone, on yohimbine-induced reinstatement. Male and female rats were trained to lever press for i.v. infusions of cocaine (0.4 mg/kg). Following a 14-day maintenance period, cocaine solutions were replaced with saline, and rats were allowed to extinguish lever pressing. Subsequently, rats were administered saline, yohimbine (2.5mg/kg), or allopregnanolone (15 mg/kg)+yohimbine (2.5mg/kg) priming injections on separate days using a within-subjects reinstatement procedure. The results indicated that females were more resistant to extinction than male rats and that both groups reinstated cocaine seeking following injections of yohimbine; however, female rats responded more than males to yohimbine-priming injections. Additionally, allopregnanolone blocked yohimbine's potentiating effect on responding in females but not males. These results suggest that females may be more sensitive than males to stress-induced reinstatement of cocaine-seeking behavior, and the progesterone metabolite, allopregnanolone, offers protection against this vulnerability. PMID- 20005643 TI - Expression from baculovirus and serological reactivity of the nucleocapsid protein of dolphin morbillivirus. AB - The nucleocapsid (N) protein of dolphin morbillivirus (DMV) was expressed from a baculovirus (Autographa californica nuclear polyhedrosis virus) vector and shown by SDS-PAGE and Western blot analysis to be about 57 kDa. Transmission electron microscopy revealed fully assembled nucleocapsid-like particles (NLPs) exhibiting the typical helical herringbone morphology. These NLPs were approximately 20-22 nm in diameter and varied in length from 50 to 100 nm. Purified DMV-N protein was used as antigen in an indirect ELISA (iELISA) and shown to react with rabbit and human antisera to measles virus (MV) and dog sera with antibodies to canine distemper virus (CDV). The iELISA was used for the demonstration of morbillivirus antibodies in the serum of cetaceans and manatees, showing potential as a serological tool for the mass screening of morbillivirus antibodies in marine mammals. PMID- 20005644 TI - Detection of hepatitis E virus in samples of animal origin collected in Hungary. AB - Hepatitis E virus (HEV) is an enterically transmitted human pathogen. HEV infections are mainly associated with acute, self-limited, icteric hepatitis with an average mortality rate of 1%. Animal reservoirs are considered to play an important role in the maintenance of the virus and in the spread of HEV to humans. HEV-induced seroconversion was described in several species, however clinical hepatitis in animals has not been observed to date. HEV strains from animals are genetically closely related to human HEV isolates, which supports the opinions on the zoonotic transmission of the virus. In this expansive study the occurrence of HEV was investigated in Hungarian wild and domesticated animal samples. HEV RNA was detected by reverse transcription-polymerase chain reaction in liver samples of wild boars, roe deer, and deer. The investigations of domestic swine samples detected HEV in 39% of the investigated Hungarian pig farms. Simultaneous investigation revealed no definite difference between liver and faeces samples of domestic pigs in the frequency of HEV positivity. The highest (36%) incidence of HEV infection was found among the 11-16-week-old pigs. Samples from domestic cattle and rodents collected in pig farms, forests and meadows were tested negative for HEV RNA. Phylogenetic analysis of partial sequences amplified within the ORF1 and ORF2 regions of selected strains revealed that the detected viruses belong to three subgroups of the third genogroup of HEV, and are closely related to human and swine HEV strains detected in different countries. The investigations revealed widespread distribution of HEV in Hungarian wild ungulate and domesticated swine populations, with considerable genetic diversity among the strains. PMID- 20005645 TI - Age susceptibility of red deer (Cervus elaphus) to paratuberculosis. AB - The aim of this study was to measure the relative susceptibility of three age classes of red deer (Cervus elaphus) to the various manifestations of infection and disease states due to Mycobacterium avium subsp. paratuberculosis (MAP) after experimental oral challenge. Three groups of seronegative female deer (30 three month-old weaners, 20 fifteen-month-old yearlings and 20 adults) received four oral doses of approximately 10(9) colony forming units (cfu) of a bovine strain of MAP. They were monitored for 50 weeks by weighing, blood sampling for immunological assays, skin testing and faecal culturing. Clinically affected animals were promptly euthanised and the remaining deer were killed at the end of the study. Necropsies were carried out and samples of intestine and associated lymph nodes were taken for culture and histopathology from all deer. Ten weaners developed clinical paratuberculosis and were euthanased 20-28 weeks post challenge (pc). No clinical cases occurred in the yearlings or adults. All 10 clinically affected weaners had severe gross and histopathological lesions typical of paratuberculosis (Johne's disease). At slaughter, gross lesions were seen in the jejunal lymph nodes of 8/17 weaners, 2/19 yearlings, and 0/20 adults. MAP was cultured from samples of the intestine and/or lymph nodes from all 10 clinical cases and from 16/17 weaners, 19/19 yearlings and 18/20 adult hinds at slaughter. Lesion Severity Scores of deer slaughtered 50 weeks pc averaged 4.9, 3.5 and 1.1 for the weaner, yearling and adult groups, respectively. At some time over the course of the trial, 24/28 weaners were antibody positive and immediately prior to slaughter, 13/17 weaners, 15/19 yearlings and 3/20 hinds were antibody positive. There is a strong age-related resistance against clinical disease and subclinical disease, but not to infection with MAP, after heavy oral challenge. PMID- 20005646 TI - Morbillivirus infection in free-ranging Atlantic bottlenose dolphins (Tursiops truncatus) from the Southeastern United States: seroepidemiologic and pathologic evidence of subclinical infection. AB - From 2003 to 2007, sera (n=234) from free-ranging Atlantic bottlenose dolphins (Tursiops truncatus) inhabiting two southeast Atlantic estuarine regions, the Indian River Lagoon (IRL), FL and Charleston, SC (CHS) were tested for antibodies to cetacean morbilliviruses as part of a multidisciplinary study of individual and population health. Positive morbillivirus titers were found on initial capture in 12 of 122 (9.8%) IRL dolphins in the absence of an epizootic. All CHS dolphins were seronegative. Positive fluctuating morbillivirus titers and seroconversion were found in IRL dolphins. Seropositivity was detected in dolphins 8-13 years of age as well as in dolphins that were alive during the 1987 1988 epizootic. During the study period, pathologic and immunohistochemical findings from stranded IRL dolphins (n=14) did not demonstrate typical morbillivirus-associated lesions or the presence of morbillivirus antigen. The findings suggest that morbillivirus infections are occurring in the absence of widespread mortality in IRL dolphins. PMID- 20005647 TI - Dydrogesterone in threatened miscarriage: a Malaysian experience. AB - INTRODUCTION: Threatened miscarriage is a common problem during pregnancy. METHODS: The aim of this prospective, open, randomised study was to determine whether dydrogesterone was more effective than conservative management alone in preventing miscarriage in women with vaginal bleeding up to week 16 of pregnancy. Women were excluded if they had a history of recurrent miscarriage. A total of 191 women were randomised to dydrogesterone (40 mg stat followed by 10mg twice daily) or conservative management (control group). The treatment was considered successful if the pregnancy continued beyond 20 weeks of gestation. RESULTS: The success rate in the dydrogesterone group was statistically significantly higher than that in the control group (87.5% vs. 71.6%; p<0.05). Miscarriage occurred in 12.5% of women in the dydrogesterone group compared with 28.4% in the control group (p<0.05). There were no differences between the groups with regard to the incidence of Caesarean section, placenta praevia, antepartum haemorrhage, preterm labour (weeks 28-36), pregnancy-induced hypertension or low birth weight (<2500 g) babies. There were no intrauterine deaths or congenital abnormalities in either group. CONCLUSION: Compared with conservative management, dydrogesterone had beneficial effects on maintaining pregnancy in women with threatened miscarriage. PMID- 20005648 TI - Hormone replacement therapy (HRT), breast cancer and tumor pathology. AB - Within an average observation period of 5-6 years, several clinical trials reported an increased risk of breast cancer due to hormone replacement therapy (HRT). However, it remains disputable, whether the increased rate of breast cancers detected within the given time frame is indeed due to newly induced tumors and thus constitutes HRT-initiated primary breast cancers. Onco pathologically speaking it appears more likely that HRT stimulates the growth of already existing small tumor nests which - due to their small size - would otherwise go undiagnosed. The major arguments are: In summary, HRT is hence more likely to be a tumor promoter than a de novo-inducer of breast cancers. PMID- 20005649 TI - The population genetic structure of North American Lucilia sericata (Diptera: Calliphoridae), and the utility of genetic assignment methods for reconstruction of postmortem corpse relocation. AB - Lucilia sericata (Diptera: Calliphoridae) is a common urban blowfly, with a worldwide distribution. It is among the most important forensic insects, and a major veterinary pest. A previous amplified fragment length polymorphism (AFLP) survey of Phormia regina, another blowfly, found that the North American population structure results from the fact that flies trapped together at a bait are predominantly comprised of related individuals. We report here a similar population genetic pattern for L. sericata in North America based on AFLP genotypes with 249 loci. A STRUCTURE analysis found no population structure on a geographic scale, and analysis of molecular variance found a moderate amount of variation attributed to samples (adults collected at the same bait at the same time, Phi(SC)=20%, P=0.001). A Mantel test found a negligible correlation between geographic and genetic distances (R(2)=0.0063, P=0.02). The mean relative relatedness coefficient for every sample was positive (mean R=0.2486+/-0.18). Gravid females in a sample, those likely to oviposit on the same corpse, showed a pattern of relatively high relatedness similar to the total survey. Therefore, this pattern of local relatedness is likely to occur with larvae in a corpse, and if so it might support a genetic test for inferring the postmortem relocation of a corpse. This is because a larva may fall from the body at the original scene as it is moved. Connecting such a "stray" larva to the larval population in a corpse would provide powerful evidence that the corpse had been at both locations. Assignment tests resulted in a 96% success rate of assigning L. sericata individuals to their samples of origin. PMID- 20005650 TI - Unidentified bodies and human remains: an Italian glimpse through a European problem. AB - The identification of cadavers (the main activity of forensic odontologists and anthropologists) is a crucial issue in forensic pathology, but the official entity of this problem is still poorly known in most countries, apart from a few American reports. In this article the authors present a descriptive study of unidentified decedents over a 14-year period (1995-2008) in Milan. The number of cadavers or human remains arriving at the morgue with no identity amounts to 454 3.1% of all autopsies at the Institute of Legal Medicine, with a mean of 32 unidentified subjects every year; 62% reached a positive identification in a period of time ranging from a few days to 10 years. 17% on an average remain unidentified. Most identification processes involved forensic odontology and anthropology. This study aims at revealing the problem and hopefully may provide some food for thought for forensic pathologists, anthropologists and odontologists so that they may focus on this issue and on possible solutions in their countries. PMID- 20005651 TI - Studies on 1-(2-phenethyl)-4-(N-propionylanilino)piperidine (fentanyl) and related compounds VII. Quantification of alpha-methylfentanyl metabolites excreted in rat urine. AB - The use of chemically modified controlled drugs, called designer drugs, is widespread internationally. In the 1980s, the dominant drugs of abuse were modifications of fentanyl formed by methylation of both the alpha-position of its phenethyl group (alpha-methylfentanyl) and the 3-position of its piperidine ring (3-methylfentanyl). Numerous analytical methods for fentanyl and its analogues, and many studies of its metabolism and major metabolites, have been reported. However, minor metabolites that reflected injection of the original compound were not included in these studies. Recently, structures of four novel and minor metabolites that reflect alpha-methylfentanyl have been reported. This study reports excretion amounts of these compounds for 96 h following peroral injection to rats of 3mg/day and urine collection every 24h. Major metabolites were the same as for fentanyl, with approximately 24% of alpha-methylfentanyl excreted as nor-fentanyl and 15% as omega, omega-1 hydroxypropiony nor-fentanyl up to 72 h post-injection. The novel metabolites were completely excreted within 48 h of injection and composed 2-3% of the total metabolite pool. The major metabolite nor-fentanyl was detected up to 72 h after injection. PMID- 20005652 TI - Hypoglossal hypoplasia and hyperplasia of the area postrema following perinatal hypoxic brain damage. AB - We report here an autopsy case of perinatal hypoxic brain damage showing hypoglossal hypoplasia and hyperplasia of the area postrema (AP) in the medulla oblongata. A 16-year-old girl who suffered from severe psychomotor developmental delay, epilepsy and tongue fasciculation, was shown by pathology to have a medullary change, as well as tongue atrophy and severe sclerotic changes in the cerebrum and cerebellum. Moderate to severe neuronal loss and gliosis were found in the brainstem. But neurons were preserved in the trigeminal nuclei, abducens nucleus and dorsal vagal nucleus. We performed a preliminary immunohistochemical analysis of sections of the medulla oblongata in our case, normal controls and disease controls with perinatal hypoxic ischemic encephalopathy (HIE). The normal and disease controls showed neither hypoglossal hypoplasia nor AP hyperplasia. The combination of hypoglossal hypoplasia and AP hyperplasia is unique and intriguing, and further analysis of the AP is required to understand developmental brain disorders. PMID- 20005653 TI - Correlation of 18F-fluorodeoxyglucose uptake on positron emission tomography with Ki-67 index and pathological invasive area in lung adenocarcinomas 30 mm or less in size. AB - BACKGROUND: (18)F-fluorodeoxyglucose positron emission tomography (FDG-PET) is commonly used to distinguish benign from malignant lesion. Recently, maximum standardized uptake value (SUVmax) on FDG-PET has found to have prognostic value. We examined the relationship between SUVmax and proliferative activities as indicated by maximum diameter of tumor opacity on mediastinal-window images (TOM), Ki-67 index, and diameter of the pathological invasive area in lung adenocarcinomas 5mm was determined as 2.15 by ROC analysis, with sensitivity of 88.3% and specificity of 84.6%. CONCLUSIONS: SUVmax correlated significantly with Ki-67 index and diameter of the pathological invasive area. The present results suggest the potential role of FDG-PET in predicting adenocarcinomas with invasive characteristics. PMID- 20005654 TI - Proposal to assign Aeromonas diversa sp. nov. as a novel species designation for Aeromonas group 501. AB - The Aeromonas group 501, also named Aeromonas sp. HG13, is taxonomically close to A. schubertii. Results obtained in previous studies, including DNA-DNA hybridization and DNA fingerprinting, suggest that Aeromonas group 501 could constitute a different Aeromonas species. In this work we have performed a polyphasic study with the two strains comprising the Aeromonas sp. HG13 in order to propose a formal species name. They could be differentiated from A. schubertii by the indole and lysine decarboxylase tests and the utilization of l-lactate. Phenotypically, both strains were also easily separated from the other Aeromonas species. Sequence analysis of the 16S rRNA gene showed high sequence similarities (>97%) between Aeromonas group 501 and all Aeromonas species. Nevertheless, sequence divergences of cpn60, dnaJ, gyrB and rpoD genes were higher than the intraspecific threshold values established for each gene (3.5%, 3.3%, 2.3% and 2.6%, respectively), while sequence divergences between strains CDC 2478-85(T) and CDC 2555-87 were low (0.6-1.1%). The DNA G+C content of the type strain was 62.2mol%. Phenotypic and genotypic evidence strongly suggests that the Aeromonas group 501 is a novel species of the genus Aeromonas, for which the name Aeromonas diversa sp. nov. is proposed. The type strain is CDC 2478-85(T) (=CECT 4254(T)=ATCC 43946(T)=LMG 17321(T)). PMID- 20005655 TI - Logical puzzles and scientific controversies: the nature of species, viruses and living organisms. AB - In the past, biologists believed that species were stable and permanent entities and they viewed them as natural kinds which, like the chemical elements, exist in nature independently of any human conceptualization. After Darwin, biologists came to accept that species were the products of evolution and natural selection and were not immutable natural kinds. Different definitions of the species category are discussed, in particular the concept of cluster class as a family resemblance concept. In order to resolve what has become known as the species problem, it is necessary to distinguish between species as concrete entities and species as abstract entities. A species can be an abstract concept but the concept also refers to specific objects in space and time, namely the organisms studied by biologists. The nature of viruses is discussed and the difference between a virus and a virus particle is emphasized. The category "virus species" is defined as a cluster class and the task of defining a virus species is distinguished from the task of identifying the members of a species using diagnostic criteria. Such an identification is feasible only when the species taxon has been established beforehand on the basis of several genotypic and phenotypic characters. It is argued that viruses are genetic parasites rather than living organisms and that they have no place in the tree of life. This interpretation is based on an analysis of the properties of living agents and living organisms. PMID- 20005657 TI - Determination of polar alkylating agents. PMID- 20005656 TI - Enhanced distribution and extended elimination of glycyrrhetinic acid in mice liver by mPEG-PLA modified (mPEGylated) liposome. AB - A rapid and simple method of high-performance liquid chromatography with UV detector for the quantification of glycyrrhetinic acid (GA) in mice plasma and tissues has been developed and validated. With the established assay method, the pharmacokinetic profiles and tissue distribution of GA in different formulations are compared in mice after intravenous administration of the drug (25mg/kg). The results showed that mPEG-PLA modified (mPEGylated) GA liposome (PL-GA) significantly prolonged the mean residence time (MRT) of GA in mice plasma and liver (MRT: 0.43+/-0.13 and 1.72+/-0.11h, respectively) than the normal GA liposome (L-GA) (MRT: 0.23+/-0.01 and 1.07+/-0.31h, respectively) and GA sodium injection (S-GA) (MRT: 0.13+/-0.01 and 0.95+/-0.08h, respectively). Moreover, PL GA specifically increased GA uptake in liver (AUC(0-infinity,)(liver) value of 1.6-fold and 1.3-fold higher than that for S-GA and L-GA, respectively) and reduced its distribution into other tissues after dosing. Due to these pharmacokinetic properties, it may be promising to develop PL-GA further as a new pharmaceutical preparation for GA on the treatment of various chronic hepatic diseases. PMID- 20005658 TI - Determination of duloxetine hydrochloride in the presence of process and degradation impurities by a validated stability-indicating RP-LC method. AB - A stability-indicating gradient reverse phase liquid chromatographic purity and assay method for duloxetine hydrochloride (DUH) was developed and validated. DUH was subjected to the stress conditions and it is sensitive towards oxidative, acid and hydrolytic degradation. Successful separation of DUH from its two process impurities and one degradation impurity formed under stress conditions was achieved on a Symmetry C18, 250x4.6mm, 5microm column using a gradient mixture of solvent A (0.01M potassium dihydrogen orthophosphate having 0.2% triethyl amine, pH adjusted to 2.5 with orthophosphoric acid) and solvent B (20:80 v/v mixture of acetonitrile and methanol). The flow rate is 1ml/min and the detection wavelength is 230nm. The mass balance was found to be in the range of 99.2-99.7% in all the stressed conditions. PMID- 20005659 TI - Thermal injury associated with propylene. PMID- 20005660 TI - Bilateral acute angle-closure glaucoma in a migraine patient receiving topiramate: a case report. AB - BACKGROUND: Patients with migraine headaches are frequently prescribed topiramate to treat their condition. CASE REPORT: We present a case of bilateral acute angle closure glaucoma occurring 2 days after topiramate therapy was increased for symptoms related to migraine. CONCLUSION: Acute angle-closure glaucoma secondary to topiramate is an uncommon but serious adverse reaction that may result in severe morbidity such as permanent visual loss if not recognized in a timely manner. Treatment differs from primary acute angle-closure glaucoma in that discontinuation of topiramate is necessary for the glaucoma to resolve. PMID- 20005661 TI - Is there any role for intravenous antivenom for snake venom ophthalmia? PMID- 20005662 TI - Evaluation of sensory loss to the traumatized arm. PMID- 20005664 TI - Traumatic shoulder pain. PMID- 20005663 TI - Geography and travel distance impact emergency department visits. AB - BACKGROUND: Little has been written about the geographic basis of emergency department (ED) visits. OBJECTIVE: The objective of this study is to describe the impact of geography on ED visits. METHODS: A retrospective analysis was conducted of ED visits during a 1-year period at a single institution using spatial interaction analysis that models the pattern of flow between a series of origins (census block groups) and a destination (ED). Patients were assigned to census block groups based upon their verified home address. The study hospital is the only Level I trauma, pediatric, and tertiary referral center in the area. There are 11 other hospitals with EDs within a 40-mile radius. Each patient visit within this radius, including repeat visits, was included. Patients with an invalid home address, a post office box address, or those who lived outside a 40 mile radius were excluded. ED visits per 100 population were calculated for each census block group. RESULTS: There were 98,584 (95%) visits by 63,524 patients that met study inclusion criteria. Visit rates decreased with increasing distance from the ED (p < 0.0001). Nineteen percent of patients lived within 2 miles, 48% within 4 miles, and 92% within 12 miles of the ED. The Connecticut border, 7 miles south of the ED (p < 0.0001), the Connecticut River, 1 mile west of the ED (p < 0.0001), and the presence of a competing ED within 1 mile (p < 0.0001) negatively impacted block group ED visit rates. Travel distance was related to the percentage of visits that were high acuity (p < 0.0001), daytime (p < 0.01), or resulted in admission (p < 0.0001). CONCLUSIONS: Geography and travel distance significantly impact ED visits. PMID- 20005665 TI - Pumpkin seed bezoar initially suspected as child abuse. PMID- 20005666 TI - Coup and contrecoup head injury resulting in anosmia. PMID- 20005667 TI - Compartment syndrome of the foot after calcaneal fracture. AB - BACKGROUND: Compartment syndrome of the foot as a result of a calcaneal fracture has received only occasional consideration in the recent Emergency Medicine literature, yet it remains a challenging diagnosis to make. The devastating consequences of untreated compartment syndrome of the foot include clawing of the lesser toes, stiffness, chronic pain, motor weakness, neurovascular dysfunction, and fixed deformities of the foot. In addition to decreased quality of life, this also leads to lost time at work and lost wages. Calcaneal fractures can lead to devastating long-term disability that is often permanent and life-altering for patients suffering from this injury. Approximately 10% of patients with these fractures develop compartment syndrome of the foot. The pathogenesis of calcaneal fractures is well recognized, and the surgical treatment techniques continue to evolve. OBJECTIVES: The objectives of this case report are to increase understanding of the pathophysiology of compartment syndrome and its short- and long-term consequences, to improve the ability to diagnose compartment syndrome, and to emphasize the need for emergent surgical treatment. CASE REPORT: A 37-year old man sustained an isolated comminuted, extra-articular calcaneus fracture that resulted in compartment syndrome of the foot. The diagnosis required measurement of several compartments in the foot. He subsequently received emergent operative decompression and experienced a positive outcome. CONCLUSION: Diagnosis of compartment syndrome of the foot is a clinical one, and diagnostic tools such as radiographic imaging and compartment pressure monitoring can help confirm the diagnosis. It is also important to understand the long-term sequelae of this injury and to involve a specialist early in the decision-making and treatment process. PMID- 20005668 TI - What is recovery? Revisiting the Betty Ford Institute Consensus Panel definition: The Betty Ford Consensus Panel and Consultants. PMID- 20005669 TI - Proteomic analysis of atherosclerotic plaque. AB - Proteins play a fundamental role in the formation and progression of plaque, but proteomic analysis of plaque as a whole is difficult, due to its heterogeneous cellular composition and an abundance of plasma proteins. Several approaches to this problem are reported in the literature; they include proteomic analysis of vascular tissues, analysis of proteins released by normal and pathological arterial walls, proteomic analysis of vascular cells and proteomic analysis of blood. In a previous study, we proposed a new strategy for studying of proteome of plaque, which permits to select the proteins exclusive to plaque by the constructing of a reference synthetic gel. In the present work, we matched the spots of the reference synthetic gel with the spots of a pool of carotid plaque, in order to select only spots exclusive to plaque from the 2-dimensional electrophoresis of the pool of plaque. We selected some spots between those exclusive and identified them by mass spectrometry. Some proteins identified are involved in transport, others take part in elimination of toxic radicals, others are metabolic enzymes or structural proteins. This study represents an example of application of the new approach which we have proposed: the reference gel of proteome of plaque permits to select, on every sample of interest, only the spots exclusive to plaque; once selected, spots can be identified by mass spectrometry and, being typical of plaque composition, could represent novel markers of lesions and vascular risk. PMID- 20005670 TI - Optimal node perturbation in linear perceptrons with uncertain eligibility trace. AB - Node perturbation learning has been receiving much attention as a method for achieving stochastic gradient descent. As it does not require direct gradient calculations, it can be applied to a reinforcement learning framework. However, in conventional node perturbation learning, the residual error due to perturbation is not eliminated even after convergence. Using infinitesimal perturbations suppresses the residual error, but such perturbations are less robust against uncertainty and noise in an eligibility trace, which is a memory of perturbation and input. We derive an optimal parameter schedule for node perturbation learning used with linear perceptrons with uncertainty in the eligibility trace. Our adaptive learning rule resolves the trade-off between robustness against the uncertainty and residual error reduction. The results obtained will be useful in designing learning rules and interpreting related biological knowledge. PMID- 20005671 TI - Evolutionary swarm neural network game engine for Capture Go. AB - Evaluation of the current board position is critical in computer game engines. In sufficiently complex games, such a task is too difficult for a traditional brute force search to accomplish, even when combined with expert knowledge bases. This motivates the investigation of alternatives. This paper investigates the combination of neural networks, particle swarm optimization (PSO), and evolutionary algorithms (EAs) to train a board evaluator from zero knowledge. By enhancing the survivors of an EA with PSO, the hybrid algorithm successfully trains the high-dimensional neural networks to provide an evaluation of the game board through self-play. Experimental results, on the benchmark game of Capture Go, demonstrate that the hybrid algorithm can be more powerful than its individual parts, with the system playing against EA and PSO trained game engines. Also, the winning results of tournaments against a Hill-Climbing trained game engine confirm that the improvement comes from the hybrid algorithm itself. The hybrid game engine is also demonstrated against a hand-coded defensive player and a web player. PMID- 20005672 TI - Parapharyngeal space tumors: surgical approaches in a series of 13 cases. AB - Tumors originating in the parapharyngeal space are rare; they comprise approximately 0.5% of head and neck tumors. Most (70-80%) are benign and the most frequent origins are salivary and neurogenic. The aim of this study is to present the surgical procedures used for the treatment of 13 patients with parapharyngeal space tumors; 11 of them were suffering from benign tumors (the most frequent being pleomorphic adenoma; 8 cases) and 2 from malignant lesions. The following surgical approaches were used: intraoral (2 cases), transcervical (4 cases) and transmandibular (7 cases) with different types of mandible osteotomies. The type of surgical approach was dictated by the type of the lesion (malignant or benign), the exact location, the size, the vascularity and the relation of the tumor to the neck neurovascular bundle. In all cases the selected surgical approach allowed the complete resection of the tumor, obtaining clear margins in cases of malignancy, without adding to the patient's preoperative morbidity. It was concluded that the surgical approach to the parapharyngeal space tumors must be adjusted to the tumor characteristics and be as wide is necessary to achieve its complete removal with safety. PMID- 20005673 TI - Maxillary infiltration anaesthesia by ropivacaine for upper third molar surgery. AB - The main purpose of this study was to assess the clinical efficacy and haemodynamic effects of ropivacaine for infiltration anaesthesia in patients undergoing surgical removal of upper third molars. The safety profile of ropivacaine was also studied by investigating the maximal venous plasma concentration of ropivacaine and the reactivity to ropivacaine of isolated human infraorbital arteries. Ropivacaine in concentrations of 0.5, 0.75 and 1% achieved dose-dependent parameters of maxillary infiltration aneasthesia, clinically relevant in concentrations 0.75 and 1%. Postoperative needs for analgesics were observed in 67-100% of patients. Haemodynamic parameters were stable during surgery with significant changes occuring 10 min after surgery. After maxillary infiltration of 2.0 ml 1% ropivacaine, the maximum venous plasma concentration (Cmax) was 82+/-15 microg/l. On isolated human infraorbital artery, ropivacaine (10(-4)M) induced endothelium-independent contraction. This study suggests that 0.75 and 1% ropivacaine offers adequate and safe intraoperative analgesia but not successful postoperative pain control for the surgical removal of upper third molars. PMID- 20005674 TI - Computer-assisted three-dimensional surgical planning: 3D virtual articulator: technical note. AB - This study presents a computer-assisted planning system for dysgnathia treatment. It describes the process of information gathering using a virtual articulator and how the splints are constructed for orthognathic surgery. The deviation of the virtually planned splints is shown in six cases on the basis of conventionally planned cases. In all cases the plaster models were prepared and scanned using a 3D laser scanner. Successive lateral and posterior-anterior cephalometric images were used for reconstruction before surgery. By identifying specific points on the X-rays and marking them on the virtual models, it was possible to enhance the 2D images to create a realistic 3D environment and to perform virtual repositioning of the jaw. A hexapod was used to transfer the virtual planning to the real splints. Preliminary results showed that conventional repositioning could be replicated using the virtual articulator. PMID- 20005675 TI - Neural correlates of reward processing in schizophrenia--relationship to apathy and depression. AB - The present study employs a new framework to categorise the heterogeneous findings on the relationship between impaired reward processing and negative and affective symptoms of schizophrenia. Based on previous behavioural and neuroimaging studies we postulate that "wanting" (i.e. anticipation) of a reward is specifically related to apathy, whereas "liking" (i.e. hedonic impact) is related to anhedonia and depression--symptoms commonly observed in schizophrenia. Fifteen patients with schizophrenia or schizoaffective disorder treated with atypical antipsychotic drugs and fifteen healthy controls performed a probabilistic monetary incentive delay task while undergoing functional magnetic resonance imaging. At the group level we found no significant differences between patients and controls in neural activation during anticipation or receipt of a reward. However, in patients with schizophrenia specific relationships between ventral-striatal activation and symptoms were observed. Ventral-striatal activation during reward anticipation was negatively correlated with apathy, while activation during receipt of reward was negatively correlated with severity of depressive symptoms. These results suggest that the link between negative symptoms and reward anticipation might specifically relate to apathy, i.e. a lack of motivation and drive. Impaired hedonic reward processing might contribute to the development of depressive symptoms in patients with schizophrenia, but it is not directly associated with self-rated anhedonia. These results indicate the necessity of more specifically differentiating negative and affective symptoms in schizophrenia in order to understand the role of the reward system in their pathogenesis. PMID- 20005676 TI - Food-dependent anaphylaxis with serum IgE immunoreactive to dairy products containing high-molecular-weight proteins. PMID- 20005677 TI - In vitro efficacy of a synthetic all-d antimicrobial peptide against clinically isolated drug-resistant strains. PMID- 20005678 TI - Spread of vancomycin-resistant Enterococcus faecium in two haematological centres in Russia. AB - This paper describes the clonal diversity of vancomycin-resistant Enterococcus faecium isolated from patients with haematological malignancies in Russia. Pulsed field gel electrophoresis (PFGE) typing of 129 vanA-positive E. faecium strains revealed 23 independent restriction profiles with two predominant clonal types. Multilocus sequence typing (MLST) of 16 strains selected from two predominant PFGE types showed that they belong to the epidemic clonal complex (CC) 17. Tn1546 like elements of isolates were compared with the prototype element from E. faecium BM4147 by polymerase chain reaction (PCR). Four different Tn1546 types were distinguished according to structural alternations. Polymorphism in the orf1 and vanSH genes was detected. However, a significant prevalence of the prototype Tn1546 was revealed. Tn1546-like elements with the same structures were observed in strains of different PFGE types. The virulence genes esp, gelE and hyl were detected by PCR in 118 isolates (91%), 87 isolates (67%) and 35 isolates (27%), respectively. In contrast, agg and cylA genes were not found. The detection frequency of esp was higher in epidemic strains than in sporadic ones (100% vs. 56%; P<0.05). This study describes a genetically variable population of vancomycin-resistant E. faecium in two Russian haematological centres. The spread of vancomycin resistance was mostly due to the distribution of the two subclones of E. faecium CC17, enriched with the virulence marker esp. At the same time, dissemination of an altered Tn1546 also occurred. PMID- 20005679 TI - Thioridazine protects the mouse from a virulent infection by Salmonella enterica serovar Typhimurium 74. AB - When administered to mice at doses of 100microg/mouse and 200microg/mouse, thioridazine (TDZ) significantly protected animals from the lethality produced by a virulent strain of Salmonella enterica serovar Typhimurium and reduced the number of bacteria retrieved from the spleen, liver and heart blood. The protection conferred by TDZ against a virulent Salmonella infection is hypothesised to be due to a reduction in the 55kDa virulence protein of the outer membrane of the organism, as this protein is almost totally absent when the organism is exposed to the phenothiazine. It is further hypothesised that the reduction in the 55kDa virulence factor renders the organism susceptible to the action of hydrolytic enzymes of the neutrophil phagolysosome, whereas in the absence of exposure to TDZ intracellular ingestion and localisation of the phagocytosed bacterium does not result in killing owing to rapid induction of the two-step PmrA/B regulon that results in the eventual synthesis and insertion of lipid A into the nascent lipopolysaccharide layer of the outer membrane. PMID- 20005680 TI - Evaluation of antifungal therapy in a neutropenic murine model of Neoscytalidium dimidiatum infection. AB - We evaluated the efficacy of amphotericin B (1.5mg/kg/day), voriconazole (60mg/kg/day) and posaconazole (60mg/kg/day) in a murine model of systemic infection caused by Neoscytalidium dimidiatum. All the treatments were able to prolong survival and to reduce the tissue burden in the spleen and kidneys of infected mice. Neither voriconazole nor posaconazole improved the results achieved with amphotericin B. PMID- 20005681 TI - Of mice and men: innate immunity in pneumococcal pneumonia. AB - Pneumococcal pneumonia is characterised by an intense inflammatory response induced mainly by cell wall components of the bacterium. Recognition of cell wall components by Toll-like receptors (TLRs) induces intracellular signalling pathways that culminate in the activation of pro-inflammatory genes through nuclear factor kappaB (NF-kappaB). Tumour necrosis factor-alpha (TNFalpha) is one of the earliest mediators produced and induces a second wave of pro- and anti inflammatory cytokines that orchestrate the immune response. The magnitude of this response in patients with pneumococcal pneumonia is a complex network and many factors must be considered in the analysis of the cytokine production pattern. First, bacterial growth and the inflammatory response are dynamic processes, produced initially as a local phenomenon with a late systemic extension. Second, host characteristics, such as different cytokine gene polymorphisms, can cause a distinct immune response. Finally, other microorganism determinants and even the immunomodulatory effect of antimicrobials may play a role in cytokine production. Recent data on innate immunity against Streptococcus pneumoniae gathered from the murine model of pneumonia, from studies of human genetic polymorphisms associated with increased susceptibility to pneumococcal infection, and from human clinical trials are discussed. Special emphasis has been placed on the description of the chronology of the complex network of innate immunity triggered by pneumococcal infection. PMID- 20005682 TI - Swine influenza (H1N1) pneumonia during the 'herald wave' of the pandemic: no increase in bacterial pneumonia without empirical antibiotics. PMID- 20005683 TI - Do treatment and illness beliefs influence adherence to medication in patients with bipolar affective disorder? A preliminary cross-sectional study. AB - Adherence to medication is essential for achieving good outcomes for patients with bipolar affective disorder. This study tested whether treatment and illness beliefs are important predictors of adherence to medication. Results indicate that beliefs are predictive, and may be a suitable target for modification in efforts to change behaviour. PMID- 20005684 TI - Duration and adherence of antidepressant treatment (2003 to 2007) based on prescription database. AB - BACKGROUND: Many patients discontinue antidepressant therapy long before the six month minimum duration recommended for the treatment of major depression and many other diagnoses. PURPOSE: To estimate the duration of antidepressant treatment and to analyse the following factors in relation to treatment adherence: age, sex, polypharmacy and type of drug. METHODS: Retrospective cohort followed up for five years (2003-2007) based on prescription database. SELECTION CRITERIA: Users who had received at least one antidepressant prescription in 2003 and who had not received antidepressants during the previous 12 months. VARIABLES STUDIED: Age, sex, drug, polypharmacy, period of treatment, packs dispensed. Adequate adherence was defined as dispensation of medication during at least 80% of the treatment period, and compliance was defined as good when, in addition, the treatment lasted more than four months. RESULTS: Of the 7525 patients selected, 56% abandoned medication during the first four months. Men were more likely to give up medication before time than women. Good compliance was recorded in 22% of patients and was twice as frequent in patients with high levels of polypharmacy than in those with low levels (31% vs. 15.3%). Patients receiving maprotiline, venlafaxine, mirtazapine, citalopram, clomipramine and fluoxetine presented the highest percentages of good compliance. CONCLUSIONS: Only one out of five patients complied with treatment for over four months. Treatment periods were shorter in men. In chronic processes, patients receiving polypharmacy presented the best compliance. PMID- 20005685 TI - Verbal fluency as a possible predictor for psychosis. AB - BACKGROUND: Neurocognitive abnormalities are prevalent in both first episode schizophrenia patients and in ultra high risk (UHR) patients. AIM: To compare verbal fluency performance at baseline in UHR in patients that did and did not make the transition to psychosis. METHOD: Baseline verbal fluency performance in UHR-patients (n=47) was compared to match first episode patients (n=69) and normal controls (n=42). RESULTS: Verbal fluency (semantic category) scores in UHR patients did not differ significantly from the score in first episode schizophrenia patients. Both the UHR group (p<0.003) and the patient group (p<0.0001) performed significantly worse than controls. Compared to the non transition group, the transition group performed worse on verbal fluency, semantic category (p<0.006) at baseline. CONCLUSIONS: Verbal fluency (semantic category) is disturbed in UHR-patients that make the transition to psychosis and could contribute to an improved prediction of transition to psychosis in UHR patients. PMID- 20005686 TI - Mixture classification model based on clinical markers for breast cancer prognosis. AB - OBJECTIVE: Accurate cancer prognosis prediction is critical to cancer treatment. There have been many prognosis models based on clinical markers, but few of them are satisfied in clinical applications. And with the development of microarray technologies, cancer researchers have discovered many genes as new markers from the gene expression data and have further developed powerful prognosis models based on these so-called genetic biomarkers. However, the application of such biomarkers still suffers from some problems. The first one is there are a great number of genes and a few samples in the gene expression data so that it is difficult to select a unified gene set to establish a stable classifier for prognosis. The second one is that, due to the experimental and technical reasons, there are existing noises and redundancies in gene expression data, which may lead to building a prognosis predictor with poor performance. The last but not the least one is the microarray experiments are so expensive currently that it is hard to obtain abundant samples. Therefore, it is practical to develop prognosis methods mainly based on conventional clinical markers in real cancer treatment applications. This paper aims to establish an accurate classification model for cancer prognosis, in order to make full use of the invaluable information in clinical data, especially which is usually ignored by most of the existing methods when they aim for high prediction accuracies. METHODS: First, this paper gives the formal description of general classification problem, and presents a novel mixture classification model to make full use of the invaluable information in clinical data, which is similar to the traditional ensemble classification models except for putting strict constraints on the construction of mapping functions to avoid voting process. Then, a two-layer instance of the proposed model, named as MRS (Mixture of Rough set and Support vector machine), is constructed by integrating rough set and support vector machine (SVM) classification methods, in which, the rough set classifier acts as the first layer to identify some singular samples in data, and the SVM classifier acts as the second layer to classify the remaining samples. Finally, MRS is used to make prognosis prediction on two open breast cancer datasets. One dataset, denoted as BRC-1 hereafter, is a high quality, publicly available dataset of 97 breast cancer tumors of node-negative patients. The other, denoted as BRC-2 hereafter, uses baseline human primary breast tumor data from LBL breast cancer cell collection containing 174 samples. RESULTS: We have done two experiments on BRC-1 and BRC-2, respectively. In the first experiment, the BRC-1 dataset is divided into train set with 78 patients (34 ones belonging to poor prognosis group and 44 ones belonging to good prognosis group) and test set with 19 patients (12 ones belonging to poor prognosis group and 7 ones belonging to good prognosis). After trained on the train set, the MRS can correctly classify all the 12 patients with poor prognosis, and 6 of 7 patients with good prognosis in the test set. The results are better than previous researches, even better than the 70-gene based biomarkers. And in the second experiment, we construct the classifiers using BRC 2 dataset, and compare MRS with other representative methods in Weka software by 5-fold cross-validation, and comparison results show that MRS has higher prediction accuracy than those methods. CONCLUSIONS: The proposed mixture classification model can easily integrate methods with different characteristics. It can overcome the shortcomings of traditional voting-based ensemble models and thus can make full use of the information in clinical data. The experimental results illustrate that our implemented MRS classifier can predict the breast cancer prognosis more accurately than previous prognostic methods. PMID- 20005688 TI - Electro-acupuncture promotes differentiation of mesenchymal stem cells, regeneration of nerve fibers and partial functional recovery after spinal cord injury. AB - In order to improve the structure and function of acute spinal cord injury, the present study investigated the effect of electro-acupuncture (EA) on the differentiation of mesenchymal stem cells (MSCs) and the regeneration of nerve fibers in transected spinal cord of rats. The differentiation of MSCs into neuron like cells and neuroglial cells and regeneraton of 5-hydroxytrptamine (HT) nerve fibers in the injured site of spinal cord were assessed after treatment with EA, MSCs transplantation, and EA plus MSCs transplantation. Compared with the control and MSCs groups, the content of endogenous neurotrophin-3 (NT-3) in the injured site and nearby tissues was increased in EA and EA+MSCs group. The differentiation of MSCs into neuronal-like cells and oligodendrocyte-like cells and number of 5-HT positive nerve fibers in the injured site were enhanced in the EA+MSCs group. Basso, Beattie, Bresnahan score of the paralyzed hindlimbs was highest in the EA+MSCs group. The present study demonstrates that electro acupuncture can promote the differentiation of MSCs and regeneration of nerve fibers in injured spinal cord through induction of endogenous NT-3, and that combination of EA and MSCs transplantation can improve partial function of paralyzed hindlimbs. PMID- 20005687 TI - Relative contribution of individual oxidized components in ox-LDL to inhibition on endothelium-dependent relaxation in rat aorta. AB - BACKGROUND AND AIM: Oxidized low-density lipoprotein (ox-LDL) causes atherosclerosis and endothelial dysfunction. No study up to the present date has examined the relative contribution of all the oxidized components in ox-LDL to inhibition on vascular function. Our aim was to investigate the effects of individual oxidized components at concentrations similar to those in ox-LDL on the impairment of endothelium-dependent relaxation in rat aorta. METHODS AND RESULTS: Rat thoracic aorta was pre-treated with lysophosphatidylcholine (LPC), cholesterol oxidized products (COPs), oxidized linoleic acid (ox-18:2) and oxidized linolenic acid (ox-18:3) at concentrations similar to those in human ox LDL. Ox-LDL as a whole caused 61% inhibition while LPC, COPs and ox-18:2 at concentrations similar to those in ox-LDL caused 12%, 24% and 19% inhibition, respectively, on endothelium-dependent relaxation, suggesting that COPs produced the most adverse effect followed by ox-18:2 and LPC in an additional way. Three COPs including 7-ketocholesterol, 7alpha-hydroxycholesterol and 7beta hydroxycholesterol showed inhibition on endothelium-dependent relaxation with E(max) being reduced to 79-87% compared with the control E(max) (95%). At Western blot analysis phosphorylation of eNOS at Ser1177 site and total eNOS were not altered by ox-LDL treatment, indicating that ox-LDL did not affect nitric oxide (NO) synthesis capacity. Ox-LDL might react directly with NO and lower NO bioavailability. CONCLUSION: The present study demonstrated the relative contribution of individual oxidized components in ox-LDL in the inhibition of endothelium-dependent relaxation in rat aorta. This inhibitory effect could be caused by the reduction of NO bioactivity. PMID- 20005689 TI - Relative gene expression in acid-adapted Escherichia coli O157:H7 during lactoperoxidase and lactic acid challenge in Tryptone Soy Broth. AB - Cross-protection of acid-adapted Escherichia coli O157:H7 against inimical stresses is mediated by the glucose-repressed sigma factor RpoS. However, many food systems in which E. coli O157:H7 occurs are complex and contain glucose. This study was aimed at investigating the contribution of acid and lactoperoxidase (LP)-inducible genes to cross-protection of E. coli O157:H7 against LP system and lactic acid (LA) in Tryptone Soy Broth (TSB). Acid-adapted and non-adapted E. coli O157:H7 were challenged to activated LP and LA at pH 4.0 and 5.0 in TSB for 6h at 25 degrees C followed by expression of acid and LP inducible genes. Acid-adapted E. coli showed cross-protection against activated LP and LA. All the acid-inducible genes tested were repressed at pH 4.0 with or without activated LP system. At pH 7.4, gadA, ompC and ompF were induced in acid adapted cells. Induction of corA occurred in non-adapted cells but was repressed in acid-adapted cells. Although acid-inducible genes were repressed at pH 4.0, high resistance of acid-adapted cells indicates that expression of acid-inducible genes occurred during acid adaptation and not the actual challenge. Repression of rpoS indicates that RpoS-independent systems contribute to cross-protection in acid-adapted E. coli O157:H7. PMID- 20005690 TI - Lethal weapons: DAP-kinase, autophagy and cell death: DAP-kinase regulates autophagy. AB - Recently, DAP-kinase was identified as one of the essential regulators of autophagy, activated by signals such as cytokines and ER stress. DAP-kinase is a tumor suppressor that mediates several cell death pathways, such as apoptosis and programmed necrosis. Likewise, functional studies suggest that DAP-kinase may direct autophagy specifically towards autophagic cell death. Several recent studies have mapped DAP-kinase function to distinct stages in autophagy signaling. These include the Beclin-1/phosphatidylinositol 3-kinase (PI(3)K) complex, which is necessary for autophagosome formation, and an interaction with the LC3 binding protein, MAP1B, which may regulate vesicle trafficking. This review will summarize the functional and mechanistic studies that have linked DAP kinase to the regulation of autophagy in general, and autophagic cell death, in particular. PMID- 20005691 TI - Illicit drugs policy through the lens of regulation. AB - The application of regulatory theory to the problem of illicit drugs has generally been thought about only in terms of 'command and control'. The international treaties governing global illicit drug control and the use of law enforcement to dissuade and punish offenders have been primary strategies. In this paper I explore the application of other aspects of regulatory theory to illicit drugs-primarily self-regulation and market regulation. There has been an overreliance on strategies from the top of the regulatory pyramid. Two other regulatory strategies--self-regulation and market regulation--can be applied to illicit drugs. Self-regulation, driven by the proactive support of consumer groups may reduce drug-related harms. Market strategies such as pill-testing can change consumer preferences and encourage alternate seller behaviour. Regulatory theory is also concerned with partnerships between the state and third parties: strategies in these areas include partnerships between police and pharmacies regarding sale of potential precursor chemicals. Regulatory theory and practice is a rich and well-developed field in the social sciences. I argue that governments should consider the full array of regulatory strategies. Using regulatory theory provides a rationale and justification to strategies that are currently at the whim of politics, such as funding for user groups. The greater application of regulatory approaches may produce more flexible and structured illicit drug policies. PMID- 20005692 TI - Does it matter how we refer to individuals with substance-related conditions? A randomized study of two commonly used terms. AB - OBJECTIVE: Stigma is a frequently cited barrier to help-seeking for many with substance-related conditions. Common ways of describing individuals with such problems may perpetuate or diminish stigmatizing attitudes yet little research exists to inform this debate. We sought to determine whether referring to an individual as "a substance abuser" vs. "having a substance use disorder" evokes different judgments about behavioral self-regulation, social threat, and treatment vs. punishment. METHOD: A randomized, between-subjects, cross-sectional design was utilized. Participants were asked to read a vignette containing one of the two terms and to rate their agreement with a number of related statements. Clinicians (N=516) attending two mental health conferences (63% female, 81% white, M age 51; 65% doctoral-level) completed the study (71% response rate). A Likert-scaled questionnaire with three subscales ["perpetrator-punishment" (alpha=.80); "social threat" (alpha=.86); "victim-treatment" (alpha=.64)] assessed the perceived causes of the problem, whether the character was a social threat, able to regulate substance use, and should receive therapeutic vs. punitive action. RESULTS: No differences were detected between groups on the social threat or victim-treatment subscales. However, a difference was detected on the perpetrator-punishment scale. Compared to those in the "substance use disorder" condition, those in the "substance abuser" condition agreed more with the notion that the character was personally culpable and that punitive measures should be taken. CONCLUSIONS: Even among highly trained mental health professionals, exposure to these two commonly used terms evokes systematically different judgments. The commonly used "substance abuser" term may perpetuate stigmatizing attitudes. PMID- 20005693 TI - Mechanical properties of incineration bottom ash: the influence of composite species. AB - The mechanical properties, including strength, deformational behavior, and wetting softening phenomena of municipal solid waste incinerator (MSWI) bottom ash are one of the major concerns for reuse applications. However, owing to the complex constituents of municipal solid waste, the properties of MSWI bottom ash are often highly variable. A series of artificial specimens with controlled chemical components were tested in this study. The test results show that the artificial bottom ash possesses the following mechanical characteristics: (1) for the strength, the frictional angles of the bottom ash under dry and saturated conditions vary from 34.8 degrees to 51.1 degrees and 26.0 degrees to 37.2 degrees, respectively; (2) for the deformation, the shear stiffness increases with the normal stress arises and degrades upon increased shearing; (3) significant wetting degradation of the strength and stiffness were observed. The multi-variable regression analysis was conducted to evaluate the associated influence of the chemical components on the strength. Among the evaluated components, Fe(2)O(3) and Al(2)O(3) are key factors; an increase in either results in higher strength at both dry and saturated conditions. The results were used to propose empirical relationships for phi(dry) and phi(sat), expressed in terms of Fe(2)O(3) and Al(2)O(3). Accordingly, a strength classification chart is proposed for engineering purposes. PMID- 20005694 TI - Environmental performance of construction waste: Comparing three scenarios from a case study in Catalonia, Spain. AB - The main objective of this paper is to evaluate environmental impacts of construction wastes in terms of the LIFE 98 ENV/E/351 project. Construction wastes are classified in accordance with the Life Program Environment Directive of the European Commission. Three different scenarios to current waste management from a case study in Catalonia (Spain) have been compared: landfilling, recycling and incineration, and these scenarios were evaluated by means of Life Cycle Assessment. The recommendations of the Catalan Waste Catalogue and the European Waste Catalogue have been taken into account. Also, the influence of transport has been evaluated. Results show that in terms of the Global Warming Potential, the most environmentally friendly treatment was recycling, followed by incineration and lastly landfilling. According to the influence of treatment plants location on the GWP indicator, we observe that incineration and recycling of construction wastes are better than landfilling, even for long distances from the building site to the plants. This is true for most wastes except for the stony types, than should be recycled close to the building site. In summary, data from construction waste of a Catalan case study was evaluated using the well established method of LCA to determine the environmental impacts. PMID- 20005695 TI - Micro-biofuel cell powered by glucose/O2 based on electro-deposition of enzyme, conducting polymer and redox mediators: preparation, characterization and performance in human serum. AB - In this study we report a new simple process to manufacture a biofuel cell consisting of a glucose oxidase (GOx) based anode and a laccase (LAc) based cathode. The process is based on the electro-deposition of the enzymes, conducting polymer and redox mediators from ultrapure water at a potential of 4V vs. AgCl/Ag. Contrary to the conventional electro-deposition from high ionic strength (buffer solution) at low applied potential (1V vs. AgCl/Ag) where only thin films could be deposited, leading to BFC with moderate power, the electro deposition from ultrapure water at 4V allows the growth of thick films leading to BFC with high power output. It was observed that the combination of polypyrrole (PPy), with ferrocenium hexafluorophosphate (FHFP) and pyrroloquinoline quinone (PQQ) to be appropriate for the electron transfer at the GOx bioanode, while the combination of polypyrrole with bis-(bipyridine)-(5-amino-phenanthroline) ruthenium bis (hexafluorophosphate)(RuPy) and 4,4-sulfonyldiphenol (SDP) to be effective for the electron transfer at the LAc biocathode. The working biofuel cell was studied at 37 degrees C in phosphate buffer solution at pH 7.4 containing 10 mM glucose and in human serum. Under these conditions, the maximum power density reached 3.1 microW mm(-2) at a cell voltage of 0.28 V in buffer solution and 1.6 microW mm(-2) at a cell voltage of 0.21 V in human serum. This study offers a new route to the development of enzymatic BFCs with high performance and provides information on enzymatic BFCs as in vivo power sources. PMID- 20005696 TI - Circuit formation and maintenance--perspectives from the mammalian olfactory bulb. AB - The rodent olfactory bulb (OB) is becoming a model system for studying how neuronal circuits develop and maintain. The OB has typical components of a sensory circuit such as ordered sensory inputs, diverse populations of interneurons, substantial neuromodulatory innervation, and projection neurons that transfer information to higher brain centers. Additionally, the OB is unique because its sensory afferents and a subset of its interneurons are continuously replaced throughout adulthood. Here, we review some recent findings on the development and maintenance of the mammalian OB circuitry. We review some of the known developmental strategies of the major OB components and discuss the ways in which the OB circuitry preserves stability in the face of ongoing changes. PMID- 20005697 TI - The development of risk groups in men with metastatic castration-resistant prostate cancer based on risk factors for PSA decline and survival. AB - AIMS OF THE STUDY: There are no known predictive factors of response in men receiving chemotherapy for metastatic castration-resistant prostate cancer (mCRPC). We investigated pre-treatment factors that predicted a 30% PSA decline (30% PSAD) within 3 months of starting chemotherapy, and assessed performance of a risk group classification in predicting PSA declines and overall survival (OS) in men with mCRPC. METHODS: In TAX327, 1006 men with mCRPC were randomized to receive docetaxel (D) in two schedules, or mitoxantrone (M), each with prednisone: 989 provided data on PSA decline within 3 months. Predictive factors for a 30% PSAD were identified using multivariable regression in D-treated men (n=656) and validated in M-treated men (n=333). RESULTS: Four independent risk factors predicted 30% PSAD: pain, visceral metastases, anaemia and bone scan progression. Risk groups (good: 0-1 factors, intermediate: 2 factors and poor: 3 4 factors) were developed with median OS of 25.7, 18.7 and 12.8 months (p<0.0001); 30% PSAD in 78%, 66% and 58% of men (p<0.001); and measurable disease response in 19%, 9% and 5% of men (p=0.018), respectively. In the validation cohort, similar predictive ability was noted for 30% PSAD, tumour response and OS. PCWG2 subtypes were also predictive but resulted in unequal grouping. C indices were 0.59 and 0.62 for 30% PSAD and OS in the validation dataset, respectively. CONCLUSIONS: Risk groups have been identified and validated that predict PSAD and OS in men with mCRPC and may facilitate evaluation of new systemic regimens warranting definitive testing in comparison with docetaxel and prednisone. Prospective validation of this classification system is needed. PMID- 20005698 TI - The formation of bio-oil from sludge by deoxy-liquefaction in supercritical ethanol. AB - A novel deoxy-liquefaction of sludge to liquid fuel (CH(2.07-1.40)O(0.30 0.06)N(0.09-0.05)S(0.02-0.0032)) was studied in supercritical ethanol. The reduction of oxygen atoms combined with hydrogen to produce H(2)O weaken the reduction of oxygen atoms in the form of CO and CO(2), and the latter process is the primary process for the reduction of oxygen during deoxy-liquefaction. The significant increment of oil and residue fractions yield was obtained in excess ethanol as extraction and polymerization process. The addition of Fe-catalyst could promote the catalytic activity of hydrogenation for free radicals, but inhibited the liquefaction conversion. Esters and acids were identified as main compounds as the decomposing, extraction and esterification process. The conversion of free radicals and intermediates to products with heterocyclics and phenol ring was promoted, and the formation of medium-boiling-point compounds without heterocyclics and phenol ring structure was inhibited at higher temperature and higher solvent filling ratio. PMID- 20005699 TI - Scots pine (Pinus sylvestris) bark composition and degradation by fungi: potential substrate for bioremediation. AB - The composition of Scots pine bark, its degradation, and the production of hydrolytic and ligninolytic enzymes were evaluated during 90 days of incubation with Phanerochaete velutina and Stropharia rugosoannulata. The aim was to evaluate if pine bark can be a suitable fungal substrate for bioremediation applications. The original pine bark contained 45% lignin, 25% cellulose, and 15% hemicellulose. Resin acids were the most predominant lipophilic extractives, followed by sitosterol and unsaturated fatty acids, such as linoleic and oleic acids. Both fungi degraded all main components of bark, specially cellulose (79% loss by P. velutina). During cultivation on pine bark, fungi also degraded sitosterol, produced malic acid, and oxidated unsaturated fatty acids. The most predominant enzymes produced by both fungi were cellulase and manganese peroxidase. The results indicate that Scots pine bark supports enzyme production and provides nutrients to fungi, thus pine bark may be suitable fungal substrate for bioremediation. PMID- 20005700 TI - Potential of four forage grasses in remediation of Cd and Zn contaminated soils. AB - A pot experiment was conducted in a greenhouse to evaluate the phytoremediation abilities of four forage grasses with respect to soil Cd and Zn pollution. High Cd pollution significantly increased the biomass of Pennisetum americanum (L.) LeekexPennisetum purpureum Schumach, showed no effect on Silphium perfoliatum Linn and significantly decreased biomass of Paspalum atratum cv. Reyan No. 11 and Stylosanthes guianensis cv. Reyan II. High Zn pollution significantly decreased biomass of all grasses. Shoot Cd extraction amounts were 624, 179, 21 and 15mug/plant for P. americanumxP. purpureum, P. atratum, S. guianensis and S. perfoliatum respectively at soil Cd concentration of 8mg/kg. The shoot Zn extraction amount for P. americanumxP. purpureum was 8189mug/plant while the other three grasses were severely intoxicated at the soil Zn concentration of 600mg/kg. P. americanumxP. purpureum and P. atratum could be useful for phytoextraction of either or both Cd and Zn pollution; S. perfoliatum could be regarded as a candidate species for phytostabilization of Cd contamination; while S. guianensis had no remediation capability. PMID- 20005701 TI - Status and prospect of lignocellulosic bioethanol production in China. AB - As a developing country with the largest population, China faces a serious challenge in satisfying its continuously increasing energy demand, especially for liquid fuel. Bioethanol production from lignocellulosic material is a potential and feasible method to solve the many problems in China, and it was supported by the Chinese government. Many research projects in China on lignocellulosics ethanol production have been carried out. After more than 30years of research, several pilot scale facilities have been constructed. This review focuses on the recent research activities and developments in lignocellulosic ethanol production during the past decade in China. As case study, a corncob biorefinery process is introduced. PMID- 20005702 TI - Composition and dynamics of sulfate-reducing bacteria during the waterflooding process in the oil field application. AB - The composition and dynamics of sulfate-reducing bacteria (SRB) during the waterflooding process of Daqing Oilfield were investigated in this study. PCR DGGE analysis indicated that the microbial communities were significantly different in each treatment unit, and the dominant members were mainly close to Clostridium sp., Thauera sp., Hydrogenophaga sp., Pseudomonas sp., Eubacterium sp. and Arcobacter sp. However, the members belonging to SRB were relatively few and mainly consisted of Desulfovibrio sp. and Desulfovibrio profundus. According to APS (adenosine-5'-phosphosulfate reductase) gene clone library and sequence analysis, it was found that SRB mainly belonged to Proteobacteria and Deltaproteobacteria. In the library, the genus Desulfovibrio of Deltaproteobacteria was about 65.2% while genus Desulfomonas, Desulfomicrobium, Desulfohalobium, Desulfonatronum, Desulfbulbus, Desulforhopalus, Desulfnema and Desulforhopalus were occupied nearly 34.8%. It was also proved that most of SRB in the oil field were enriched in the ground treatment unit and they are not in the liquid of oil well. PMID- 20005703 TI - Synthesis and evaluation of 3-amino-6-aryl-pyridazines as selective CB(2) agonists for the treatment of inflammatory pain. AB - A series of 3-amino-6-aryl-pyridazines have been identified as CB(2) agonists with high efficacy and selectivity against the CB(1) receptor. Details of the investigation of structure-activity relationships (SAR) are disclosed, which led to the identification of pyridazine analogue 35, a compound with high potency in an in vivo model of inflammatory pain. PMID- 20005704 TI - Novel thioamide derivatives as neutral CB1 receptor antagonists. AB - A novel class of cannabinoid-1 (CB1) receptor antagonists for the treatment of obesity is presented. The carboxamide linker in a set of 5,6-diaryl-pyrazine-2 amide derivatives was transformed into the corresponding thioamide, by using a one-pot synthesis. The structural series of thioamides not only showed retained CB1 potency (below 10nM), but also showed improved solubility. In addition, the neutral antagonist 2c significantly reduced body weight in cafeteria diet obese mice. PMID- 20005705 TI - Synthesis and screening of a CaaL peptide library versus FTase reveals a surprising number of substrates. AB - Proteins bearing a CaaL sequence are typically geranylgeranylated to enable their proper localization and function. We found that many of the dansyl-GCaaL peptides representing mammalian CaaL proteins can be farnesylated by FTase. This result may have important implications for prenylated protein biology. PMID- 20005706 TI - Novel substituted (Z)-5-((N-benzyl-1H-indol-3-yl)methylene)imidazolidine-2,4 diones and 5-((N-benzyl-1H-indol-3-yl)methylene)pyrimidine-2,4,6(1H,3H,5H) triones as potent radio-sensitizing agents. AB - A series of (Z)-5-((N-benzyl-1H-indol-3-yl)methylene)imidazolidine-2,4-dione (9a 9m) and 5-((N-benzyl-1H-indol-3-yl)methylene)pyrimidine-2,4,6(1H,3H,5H)-trione (10a-10i) derivatives that incorporate a variety of aromatic substituents in both the indole and N-benzyl moieties have been synthesized. These analogs were evaluated for their radiosensitization activity against the HT-29 cell line. Three analogs, 10a, 10b, and 10c were identified as the most potent radiosensitizing agents. PMID- 20005707 TI - Synthesis and biological evaluation of nitrogen-containing chalcones as possible anti-inflammatory and antioxidant agents. AB - A novel series of nitrogen-containing chalcones were synthesized by Mannich reaction and were screened for anti-inflammatory related activities such as inhibition of cyclooxygenase-2 (COX-2), trypsin and beta-glucuronidase. The antioxidant potential was demonstrated using 1,1-diphenyl-2-picryl hydrazine (DPPH) radical scavenging activity. The results of the above studies shows that the compounds synthesized were found to be effective inhibitors of above pro inflammatory enzymes, and were found to be possess moderate radical scavenging potential. Overall, the results of the studies reveal that the chalcones with N methyl piperazine methyl and piperidine methyl substitution (4c, 3b, 4d, 6b) seems to be important for inhibition of beta-glucuronidase. Whereas the chalcones with piperidine methyl substitution (8b, 7b, 7c, 6c, 4b, 3c, 3b) were observed as effective inhibitors of COX-2, while the same compounds were found to be less reactive against COX-1 as compared to COX-2. PMID- 20005708 TI - Synthesis and antiproliferating activity of iron chelators of hydroxyamino-1,3,5 triazine family. AB - We synthesized and evaluated new specific tridentate iron(III) chelators of 2,6 bis[hydroxyamino]-1,3,5-triazine (BHT) family for use in iron deprivation cancer therapy. Physical properties of BHT chelators are easily customizable allowing easy penetration through cellular membranes. Antiproliferative activity of new BHT chelators was studied on MDA-MB-231 and MiaPaCa cells and compared to a clinically available new oral iron chelator, deferasirox (DFX). The antiproliferative activity of new chelators was found to correlate with iron(III) chelation ability and some of analogs showed substantially higher antiproliferative activity than DFX. PMID- 20005709 TI - Carbonic anhydrase inhibitors. X-ray crystal studies of the carbonic anhydrase II trithiocarbonate adduct--an inhibitor mimicking the sulfonamide and urea binding to the enzyme. AB - Trithiocarbonate (CS32-) inhibits with low micromolar affinities several mammalian carbonic anhydrases, CAs, EC 4.2.1.1 [Innocenti et al., Bioorg. Med. Chem. Lett. 2009, 19, 1855]. Here we report the X-ray crystal structure of the hCA II-trithiocarbonate adduct. Trithiocarbonate is monodentately bound to the Zn(II) ion and makes several hydrogen bonds with Thr199 and two water molecules from the enzyme active site. Its binding is different from that of ureate, another small inhibitor isosteric with trithiocarbonate but somehow mimicks the binding of the SO(2)NH moiety present in the sulfonamide inhibitors and is similar to that of bicarbonate. Compounds incorporating this new zinc-binding group, CS2-, may thus lead to new classes of potent inhibitors. PMID- 20005710 TI - Non-aromatic A-ring replacement in the triaryl bis-sulfone CB2 receptor inhibitors. AB - The triaryl bis-sulfone 1 was modified by converting the aryl A-ring to a piperidine ring. The piperidine ring was further elaborated to a spirocyclopropyl piperidine moiety. The effect on CB2 binding potency, rat calcium channel affinity, and CYP 2C9 inhibition is described. PMID- 20005711 TI - 3-(3-Aryloxyaryl)imidazo[1,2-a]pyridine sulfones as liver X receptor agonists. AB - Replacement of a quinoline with an imidazo[1,2-a]pyridine in a series of liver X receptor (LXR) agonists incorporating a [3-(sulfonyl)aryloxyphenyl] side chain provided high affinity LXR ligands 7. In functional assays of LXR activity, good agonist potency and efficacy were found for several analogs. PMID- 20005712 TI - Spirodiketopiperazine-based CCR5 antagonists: Improvement of their pharmacokinetic profiles. AB - Spirodiketopiperazine-based CCR5 antagonists, showing improved pharmacokinetic profiles without reduction in antagonist activity, were designed and synthesized. We also demonstrate the anti-HIV activity of a representative compound 12, as measured in a p24 assay. PMID- 20005713 TI - Rigid spine syndrome revealing late-onset Pompe disease. AB - The authors describe a 50-year-old man who was evaluated for a rigid spine syndrome with onset at age 15, and subsequent walking difficulties. Cardiac and pulmonary functions were normal. Deltoid biopsy revealed the presence of small vacuoles and increased glycogen with Periodic Acid Schiff staining in a limited number of fibers. Acid alpha-glucosidase staining was decreased in leucocytes, and genetic analysis identified the presence of two mutations in that gene. This observation suggests that Pompe disease should be considered in the differential diagnosis of rigid spine syndrome, even in patients without respiratory involvement or with a muscle biopsy showing only mild histopathological changes. PMID- 20005714 TI - Must we always hunt for a sentinel node? PMID- 20005715 TI - Finding scaffold proteins in interactomes. PMID- 20005716 TI - The importance of internal strain as opposed to interface pressure in the prevention of pressure related deep tissue injury. AB - For pressure ulcer prevention an ambitious goal would be the establishment of a mechanical threshold for tissue damage. In the past, several researchers have sought to establish such a threshold often involving the loading time. However, they have not resulted in a unique reliable value that could be used in practice. This limitation is probably due to the focus on interface pressure. The objective of this paper is to clarify to an audience with no conventional background in mechanics, why interface pressure is not the appropriate parameter to define a damage threshold, whereas internal local deformations (strains) may prove more suitable. The paper reveals that it may be possible to identify a damage threshold for healthy skeletal muscle tissue based on local internal deformations. PMID- 20005717 TI - Materials for engineering vascularized adipose tissue. AB - Loss of adipose tissue can occur due to congenital and acquired lipoatrophies, trauma, tumor resection, and chronic disease. Clinically, it is difficult to regenerate or reconstruct adipose tissue. The extensive microvsacular network present in adipose, and the sensitivity of adipocytes to hypoxia, hinder the success of typical tissue transfer procedures. Materials that promote the formation of vascularized adipose tissue may offer alternatives to current clinical treatment options. A number of synthetic and natural biomaterials common in tissue engineering have been investigated as scaffolds for adipose regeneration. While these materials have shown some promise they do not account for the unique extracellular microenvironment of adipose. Adipose derived hydrogels more closely approximate the physical and chemical microenvironment of adipose tissue, promote preadipocyte differentiation and vessel assembly in vitro, and stimulate vascularized adipose formation in vivo. The combination of these materials with techniques that promote rapid and stable vascularization could lead to new techniques for engineering stable, vascularized adipose tissue for clinical application. In this review we discuss materials used for adipose tissue engineering and strategies for vascularization of these scaffolds. CLINICAL RELEVANCE: Materials that promote formation of vascularized adipose tissue have the potential to serve as alternatives or supplements to existing treatment options, for adipose defects or deficiencies resulting from chronic disease, lipoatrophies, trauma, and tumor resection. PMID- 20005718 TI - Calculation of joint moments following foot contact across two force plates. AB - This study aimed to quantify the effect of combining the measurements from force plates when a subject's foot comes in contact with more than one force plate. A 3 Dimensional Gait Analysis (3DGA) was performed on a subject walking barefoot. Ten gait trials (good both) were captured where both subject's feet hit a single force plate. Then 20 gait trials (two force plates) were captured where either the right or left subject's foot was in contact with two force plates at a time. Kinematics were computed with VICON Plugin Gait and kinetics with a BodyLanguage (VICON, Oxford, UK) model that allowed the combination of force plate measurements. The kinetics traces from both sets of data were compared using variance component analysis. Results suggest that effects of how the moments were calculated were at most a third of those arising from stride to stride variability. This suggests that development of automated systems for determining foot contact coupled with arrays of more and smaller force plates than are commonly used might be useful to ensure the capture of good quality kinetic data in a wide range of patients. PMID- 20005719 TI - Outcome following surgery for ophthalmic segment aneurysms. AB - Ophthalmic segment aneurysms account for about 5% of all intracranial aneurysms. Anatomical complexity of the paraclinoid region makes surgical management of aneurysms arising from the ophthalmic segment challenging. This study was carried out to assess the presenting features, complications and outcomes after surgical treatment of ophthalmic segment aneurysms. The authors retrospectively analysed the clinical records of patients with ophthalmic aneurysms treated at our Institute from January 2001 to September 2008, which constituted about 9% (78/850) of all intracranial aneurysms. Of the 78 ophthalmic segment aneurysms, six patients (8%) had giant aneurysms and 19 (24%) patients had multiple aneurysms. Fifty-six patients underwent microsurgery, with direct clipping in most. The mean age was 42 years (range 12-75 years) and the mean follow-up was 8 months (range, 2-93 months). A good outcome was achieved in 46 (83%) patients (Glasgow Outcome Scale [GOS] score 4-5) and 17% had a poor outcome (GOS score 1 3) at last follow-up. The overall complication rate was 21% (12/56), most of which were transient complications, with 3.5% (2/56) mortality. Direct microsurgical clipping remains our preferred treatment approach, whenever possible, for ophthalmic segment aneurysms. This surgery has an acceptable complication rate and leads to a good outcome in more than 80% of patients with ophthalmic aneurysms. Use of modern microsurgical instrumentation and endovascular adjuncts can further reduce the surgical morbidity associated with these vascular lesions. PMID- 20005720 TI - Brain stem cavernous malformations. AB - We retrospectively reviewed the clinical experience of 30 patients with brain stem cavernous malformations (BSCM) treated operatively and non-operatively at our hospital between 1983 and 2005 to elucidate the natural history of BSCM and the factors that affect surgical outcome. Inpatient charts, imaging studies, operative records, and follow-up results were evaluated. The average follow up was 48.5 months. Twenty-two patients (73.3%) received surgical extirpation and of these 86.4% improved or stabilized and 13.6% deteriorated with permanent or severe morbidity. There was no mortality. Size, preoperative status, and surgical timing were factors related to surgical outcome. In the non-operative group, 50% of the patients were the same or better, 25% deteriorated, and 25% died. With appropriate patient selection, resection of BSCM can be achieved with acceptable morbidity compared with the ominous natural history of these lesions. PMID- 20005721 TI - Periprocedural MRI perfusion imaging to assess and monitor the hemodynamic impact of intracranial angioplasty and stenting for symptomatic atherosclerotic stenosis. AB - We aimed to assess the clinical value of MRI perfusion imaging in the periprocedural management of intracranial atherosclerosis, analyzing if changes in mean transit time (MTT), cerebral blood volume (CBV) and cerebral blood flow (CBF) correlated with angiographic outcomes. Pre-procedural and post-procedural MRI perfusion was performed on six patients who underwent angioplasty and/or stenting for symptomatic intracranial atherosclerosis. MTT, CBV and CBF were analyzed and graded. In 83% of patients, perfusion imaging correlated with angiographic outcomes. Perfusion parameters improved to normal in two patients. Two showed marked improvement and one showed mild improvement. In one patient, the results of the post-procedural MRI perfusion prompted an angiogram, which confirmed stent occlusion. Semi-quantitative scores of MTT and CBF changed over time (p=0.05, p=0.03) whereas CBV did not change significantly (p>0.05). We conclude that MRI perfusion appears a promising technique for analyzing the impact of intracranial stenosis on cerebral hemodynamics before and after treatment. PMID- 20005722 TI - Nocardial brain abscess. AB - Nocardial infections, although rare, are challenging for clinicians to treat. The associated mortality rate remains high; such infections usually occur in immunocompromised patients who have predisposing factors such as malignancy, diabetes mellitus, malnutrition and uremia. However, there have been increasing reports of nocardial infections being observed in immunocompetent patients. Nocardial organisms are mostly isolated from plants and soil, and infection occurs most often as a result of inhalation or direct skin inoculation. Nocardial infections disseminate hematogenously from the primary location to distant end organs, including the brain, kidneys, joints and eyes. Sulfonamides are the drug of choice, based on empirical data. Given the high rate of relapse and the characteristic resistance pattern, treatment should be aggressive and continued for months, with antibiotic treatment being adjusted according to the drug sensitivity test. In our institution, there have been three documented patients with a nocardial brain abscess. All patients were treated with surgical evacuation followed by antibiotics. Here, we report on one patient and review the literature. PMID- 20005723 TI - How do the ends replicate? PMID- 20005724 TI - Characterization of non-lipid autotaxin inhibitors. AB - Autotaxin (ATX) is a member of the ecto-nucleotide pyrophosphatase/phosphodiesterase (NPP) family and is a lysophospholipase D that cleaves the choline headgroup from lysophosphatidylcholine to generate the bioactive lipid lysophosphatidic acid (LPA). Enhanced expression of ATX and specific receptors for LPA in numerous cancer cell types has created an interest in studying ATX as a potential chemotherapeutic target. Likewise, ATX has been linked to several additional human diseases including multiple sclerosis, diabetes, obesity, neuropathic pain, and Alzheimer's disease. ATX inhibitors reported to date consist of metal ion chelators, lipid-like product analogs, and non-lipid small molecules. In the current research, we examined the pharmacology of the best of our previously reported non-lipid small molecule inhibitors. Here, these six inhibitors were studied utilizing the synthetic fluorescent lysophospholipid substrate FS-3, the nucleotide substrate pNP-TMP and the endogenous substrate LPC (16:0). All six compounds inhibited FS-3 hydrolysis >or=50%, whereas only three inhibited the hydrolysis of pNP-TMP to this degree. None of the six compounds blocked LPC 16:0 hydrolysis within the desired 50% inhibition range. The most potent analog (5, H2L 7905958) displayed an IC(50) of 1.6microM (K(i)=1.9microM, competitive inhibition) with respect to ATX-mediated FS-3 hydrolysis and an IC(50) of 1.2microM (K(i)=K(i)(')=6.5microM, non competitive inhibition) against ATX-mediated pNP-TMP hydrolysis. All six inhibitors were specific for ATX as they were without affect on two additional lipid preferring NPP isoforms. PMID- 20005725 TI - 1-Indol-1-yl-propan-2-ones and related heterocyclic compounds as dual inhibitors of cytosolic phospholipase A(2)alpha and fatty acid amide hydrolase. AB - Cytosolic phospholipase A(2)alpha (cPLA(2)alpha) and fatty acid amide hydrolase (FAAH) are enzymes, which have emerged as attractive targets for the development of analgetic and anti-inflammatory drugs. We recently reported that 1-[3-(4 octylphenoxy)-2-oxopropyl]indole-5-carboxylic acid (10) and related compounds are inhibitors of cPLA(2)alpha. Since cPLA(2)alpha and FAAH possess several common structural features, we now screened this substance series together with some new derivatives for FAAH inhibition. Some of the assayed compounds proved to be selective cPLA(2)alpha inhibitors, while others showed high FAAH and moderate cPLA(2)alpha inhibitory potency. Furthermore, several derivatives were favorably active against both enzymes and, therefore, could represent agents, which have improved analgetic and anti-inflammatory qualities in comparison with selective cPLA(2)alpha and FAAH inhibitors. PMID- 20005726 TI - Development of carbohydrate-derived inhibitors of acid sphingomyelinase. AB - The acid sphingomyelinase is an emerging drug target, especially for inflammatory lung diseases. Presently, there are no directly-acting potent inhibitors available for cell-based studies. The potent inhibitor phosphatidylinositol-3,5 bisphosphate (PtdIns3,5P2) is not only unsuited for cell culture studies, but also does not provide hints for further structural improvements. In the SAR study described here, we replaced the inositolphosphate moiety by a carbohydrate derivative and the phosphatidic acid residue by an alkylsulfone ester. The resulting compound is more active than its parent compound and offers new means for further structural modification. PMID- 20005727 TI - Bisquaternary pyridinium oximes: Comparison of in vitro reactivation potency of compounds bearing aliphatic linkers and heteroaromatic linkers for paraoxon inhibited electric eel and recombinant human acetylcholinesterase. AB - Oxime reactivators are the drugs of choice for the post-treatment of OP (organophosphorus) intoxication and used widely for mechanistic and kinetic studies of OP-inhibited cholinesterases. The purpose of the present study was to evaluate new oxime compounds to reactivate acetylcholinesterase (AChE) inhibited by the OP paraoxon. Several new bisquaternary pyridinium oximes with heterocyclic linkers along with some known bisquaternary pyridinium oximes bearing aliphatic linkers were synthesized and evaluated for their in vitro reactivation potency against paraoxon-inhibited electric eel acetylcholinesterase (EeAChE) and recombinant human acetylcholinesterase (rHuAChE). Results herein indicate that most of the compounds are better reactivators of EeAChE than of rHuAChE. The reactivation potency of two different classes of compounds with varying linker chains was compared and observed that the structure of the connecting chain is an important factor for the activity of the reactivators. At a higher concentration (10(-3)M), compounds bearing aliphatic linker showed better reactivation than compounds with heterocyclic linkers. Interestingly, oximes with a heterocyclic linker inhibited AChE at higher concentration (10(-3)M), whereas their ability to reactivate was increased at lower concentrations (10(-4)M and 10(-5)M). Compounds bearing either a thiophene linker 26, 46 or a furan linker 31 showed 59%, 49% and 52% reactivation of EeAChE, respectively, at 10(-5)M. These compounds showed 14%, 6% and 15% reactivation of rHuAChE at 10(-4)M. Amongst newly synthesized analogs with heterocyclic linkers (26-35 and 45-46), compound 31, bearing furan linker chain, was found to be the most effective reactivator with a k(r) 0.042min(-1), which is better than obidoxime (3) for paraoxon-inhibited EeAChE. Compound 31 showed a k(r) 0.0041min(-1) that is near equal to pralidoxime (1) for paraoxon inhibited rHuAChE. PMID- 20005728 TI - A comparative study of two marine catfish (Siluriformes, Ariidae): Cytogenetic tools for determining cytotaxonomy and karyotype evolution. AB - The family Ariidae comprises approximately 130 catfish species on both warm temperate and tropical continental shelves around the world. The systematics of the group is problematic, with several misidentification problems. In order to better understand the evolutionary relationships in the family, the present study used a cytogenetic approach to characterize two populations of Genidens genidens and two populations of Aspistor luniscutis from the southern coast of Brazil using conventional techniques and fluorescent in situ hybridization with 18S rDNA probes. The two species had the same diploid number (2n = 56), high fundamental numbers and similar banding patterns, thereby corroborating the karyotypic homogeneity proposed for the group. Single nucleolus organizer regions (NORs) were found in the genus Genidens and multiple NORs were found in Aspistor, which are considered an important cytotaxonomic marker for this genus. Karyotypic evolution trends were hypothesized, providing a better understanding of the karyotype diversity and chromosome evolution processes. PMID- 20005729 TI - Visualising impregnated chitosan in Pinus radiata early wood cells using light and scanning electron microscopy. AB - Chitosan, a deacetylated product of an abundant naturally occurring biopolymer chitin, has been used in a range of applications, particularly in food and health areas, as an antimicrobial agent. In the work reported here Pinus radiata wood was impregnated with chitosan as an environmentally compatible organic biocide (Eikenes et al., 2005a,b) to protect wood against wood deteriorating microorganisms and to thus prolong the service life of wooden products. We developed sample preparation techniques targeted to visualise impregnated chitosan within wood tissues using light microscope and field-emission scanning electron microscope (FE-SEM). Sections were viewed with the light microscope without staining with a dye as well as after staining with the dye toluidine blue. Light microscopy was also undertaken on sections that had been stained with 1% aqueous osmium tetroxide (OsO(4)). For SEM observations, the sections were treated with OsO(4) and then examined with the FE-SEM, first in the secondary electron imaging mode (SEI) and then in the backscattered electron imaging (BEI) mode, imaging the same areas of a section in both SEI and BEI modes. The preparation techniques employed and the combined use of light and scanning electron microscopy provided valuable complementary information, revealing that chitosan had penetrated into the cavities (cell lumens, intercellular spaces) of all sizes present within wood tissues and had also impregnated early wood cell walls. The information obtained is discussed in relation to its importance in further development of chitosan formulations and refinement of impregnation technologies to optimise chitosan impregnation into and distribution within wood tissues as well as in assessing chitosan efficacy. PMID- 20005730 TI - Development of secondary standards for 223Ra. AB - Ra-223 is a bone-seeking alpha emitter currently being evaluated as a radiopharmaceutical. Concurrent with the primary standardization, NIST established that calibration factors currently used for radionuclide calibrators in the clinical setting give readings 5.7-8.7% higher than the NIST calibrated activity. This work describes the determination of calibration factors specific to dose vials and syringes. Using the calibration factors derived with standard ampoules to measure syringe activities can give readings up to 3.6% too high. PMID- 20005731 TI - EC comparison on the determination of 226Ra, 228Ra, 234U and 238U in water among European monitoring laboratories. AB - In anticipation of new European requirements for monitoring radioactivity concentration in drinking water, IRMM organized an interlaboratory comparison on the determination of low levels of activity concentrations (about 10-100 mBq L( 1)) of the naturally occurring radionuclides (226)Ra, (228)Ra, (234)U and (238)U in three commercially available mineral waters. Using two or three different methods with traceability to the International System of Reference (SIR), the reference values of the water samples were determined prior to the proficiency test within combined standard uncertainties of the order of 3%-10%. An overview of radiochemical separation and measurement methods used by the 45 participating laboratories are given. The results of the participants are evaluated versus the reference values. Several of the participants' results deviate by more than a factor of two from the reference values, in particular for the radium isotopes. Such erroneous analysis results may lead to a crucial omission of remedial actions on drinking water supplies or to economic loss by an unjustified action. PMID- 20005732 TI - Protein arginine methylation in estrogen signaling and estrogen-related cancers. AB - Estrogen signaling pathways regulate multiple cellular processes including proliferation and differentiation, and dysregulation of these pathways underlies several human pathologies. Post-translational modifications (PTMs) play an important role in estrogen signaling. This review focuses on recent findings pertinent to arginine methylation of non-histone proteins and their implications in estrogen signaling. We describe protein arginine methyltransferases and demethylases, the role of methylarginine proteins in estrogen action and crosstalk with other PTMs such as phosphorylation and lysine methylation. The relationships between various PTMs form a specific code that is likely to play an important role in hormone signaling. In addition, dysregulation of arginine methylation or of enzymes responsible for these modifications could be key events in estrogen-dependent cancers such as breast cancer. PMID- 20005733 TI - Novel neural pathways for metabolic effects of thyroid hormone. AB - The relation between thyrotoxicosis, the clinical syndrome resulting from exposure to excessive thyroid hormone concentrations, and the sympathetic nervous system remains enigmatic. Nevertheless, beta-adrenergic blockers are widely used to manage severe thyrotoxicosis. Recent experiments show that the effects of thyrotoxicosis on hepatic glucose production and insulin sensitivity can be modulated by selective hepatic sympathetic and parasympathetic denervation. Indeed, thyroid hormone stimulates hepatic glucose production via a sympathetic pathway, a novel central pathway for thyroid hormone action. Rodent studies suggest that similar neural routes exist for thyroid hormone analogues (e.g. thyronamines). Further elucidation of central effects of thyroid hormone on autonomic outflow to metabolic organs, including the thyroid and brown adipose tissue, will add to our understanding of hyperthyroidism. PMID- 20005734 TI - Intergenerational transmission of programmed effects: public health consequences. AB - Epidemiological studies have shown that the environment experienced in early life can 'programme' susceptibility to later disease. Furthermore, there is increasing evidence that these effects can be transmissible to subsequent generations through non-genomic mechanisms, with profound implications for human populations. Several mechanisms can underpin the intergenerational transmission of the programmed phenotype, including persistence of the abnormal environment across generations, maternal effects and the transmission of epigenetic information through the germline. In this review, we discuss the evidence for these mechanisms in human and animal studies and the potential importance of this field for human health. PMID- 20005735 TI - Inhibins: from reproductive hormones to tumor suppressors. AB - Inhibins are peptide hormones shown originally to be produced by the gonads to regulate the secretion of follicle stimulating hormone by pituitary gonadotropes. Although gonadotropes have been regarded as the canonical inhibin target cells, in recent years extrapituitary actions of inhibins have come into the spotlight. In particular, disruptions to the local actions of inhibins in peripheral tissues might underlie certain diseases, especially cancers of the reproductive tract. This review focuses on recent advances in the inhibin field, with a particular emphasis on the determinants of inhibin availability, mechanisms of inhibin action, and the physiological relevancy of local inhibin actions in the development and progression of reproductive cancers. PMID- 20005737 TI - A review of the herbal phosphodiesterase inhibitors; future perspective of new drugs. AB - Phosphodiesterase inhibitors (PDEIs) are a class of drugs that are widely used because of their various pharmacological properties including cardiotonic, vasodilator, smooth muscle relaxant, antidepressant, antithrombotic, bronchodilator, antiinflammatory and enhancer of cognitive function. In the recent years, interest in drugs of plant origin has been progressively increased. Some pharmacologically active substances that come from plants demonstrate PDEI activity. They mainly belong to alkaloids, flavonoids, and saponins. In this review, studies on herbal PDEI were reviewed and their possible therapeutic applications were discussed. Screening plants for PDE inhibitory activity may help to develop standardized phytotherapeutic products or find new sources for new lead structures with PDEI pharmacological activity. The studies discussed in this paper are mainly in vitro and for more reasonable and conclusive results, it is required to conduct in vivo and finally human and clinical tests. PMID- 20005736 TI - Trafficking of G-protein-coupled receptors to the plasma membrane: insights for pharmacoperone drugs. AB - G protein-coupled receptors (GPCRs) are among the most common potential targets for pharmacological design. Synthesized in the endoplasmic reticulum, they interact with endogenous chaperones that assist in folding (or can retain incorrectly folded proteins) and are transferred to the plasma membrane where they exert their physiological functions. We summarize trafficking of the gonadotropin-releasing hormone receptor (GnRHR) to the plasma membrane. The trafficking of GnRHR is among the best characterized due in part to its small size and the consequent ease of making mutant proteins. Human mutations that cause disease through the misrouting of GPCRs including GnRHR are also reviewed. Special emphasis is placed on therapeutic opportunities presented by pharmacological chaperone drugs, or pharmacoperones, that allow misrouted mutants to be routed correctly and restored to function. PMID- 20005738 TI - The expression of IL-8 and IL-8 receptors in pancreatic adenocarcinomas and pancreatic neuroendocrine tumours. AB - BACKGROUND: Inflammatory mediators influence tumour progression. IL-8 has been shown to have pro-angiogenic, mitogenic and motogenic effects and several studies have demonstrated the expression of IL-8 by various human pancreatic cancer cell lines. METHODS: The expression of IL-8 and IL-8 receptors was studied in 52 pancreatic adenocarcinomas and 52 pancreatic neuroendocrine tumours using immunohistochemistry. The expression of IL-8 and IL-8 receptors was also assessed in eight pancreatic adenocarcinomas and seven neuroendocrine tumours in comparison to normal pancreatic tissue using real time quantitative PCR (qRT PCR). RESULTS: Immunohistochemical analysis of the expression of IL-8, IL-8RA and IL-8RB in 52 pancreatic adenocarcinomas demonstrated expression in 25%, 75% and 79% of pancreatic adenocarcinomas, respectively. There was no statistically significant correlation between expression and tumour grade and stage for any of the three antigens. IL-8, IL-8RA and IL-8RB expression was detected in 21%, 63% and 92% of 52 pancreatic neuroendocrine tumours. There was no statistically significant correlation between expression and tumour grade for any of the three antigens. Using qRT-PCR, the expression of each of IL-8, IL-8RA and IL-8RB mRNA was increased in 75% of pancreatic adenocarcinomas. IL-8, IL-8RA and IL-8RB mRNA expression was also increased in 57%, 43% and 29% of pancreatic neuroendocrine tumours. Quantitatively, there was a significant increase in expression level of IL-8 in tumours of both types in comparison to normal pancreatic tissue (38.5 fold in adenocarcinomas and 43.9-fold in neuroendocrine tumours). There was also increased expression of IL-8RA in both tumour types, with higher levels in adenocarcinomas, 2.7-fold and neuroendocrine tumours, 1.7-fold. IL-8RB was slightly increased in adenocarcinomas in comparison to normal pancreas (1.4 fold), but the expression was decreased in neuroendocrine tumours compared with normal pancreas (0.9-fold). CONCLUSION: This is the first study to show that IL-8 and IL-8 receptors are upregulated in both pancreatic adenocarcinomas and neuroendocrine tumours, and indicate this signalling pathway may modulate tumour behaviour through autocrine and/or paracrine loops. PMID- 20005739 TI - Serum interleukin-6 and C-reactive protein are markedly elevated in acute decompensated heart failure patients with left ventricular systolic dysfunction. AB - Cytokines play important roles in heart failure (HF). We examined whether cytokine levels are different in acute decompensated heart failure (ADHF) patients between with left ventricular systolic dysfunction (LVSDF) and with preserved LV ejection function (PLVEF). We studied 81 HF patients who were admitted to our hospital with acute decompensation. They were divided into two groups: LVSDF (LVEF)<45% and PLVEF (LVEF45%). Serum interleukin-6 (IL-6), highly sensitive C-reactive protein (hsCRP), tumor necrosis factor alpha (TNF-alpha), and IL-18 and plasma brain natriuretic peptide (BNP) were measured on admission and at discharge. On admission, IL-6 and hsCRP were higher in LVSDF than in PLVEF. IL-6 and hsCRP decreased after treatment in LVSDF, but not in PLVEF, while plasma BNP levels decreased in both HF with treatment. There was no difference in TNF-alpha or in IL-18 level between LVSDF and PLVEF, and they did not change after treatment in either group. In conclusion, cytokine profiles were different in ADHF between those with LVSDF and PLVEF. Activation of IL-6-hsCRP pathway may play a specific role in ADHF with LVSDF. PMID- 20005740 TI - H/D exchange kinetics: experimental evidence for formation of different b fragment ion conformers/isomers during the gas-phase peptide sequencing. AB - Electrospray ionization (ESI) Fourier transform ion cyclotron resonance mass spectrometry (FT-ICR MS) combined with H/D exchange reactions was utilized to explore the existence of different b(5)(+) and b(4)(+) fragment ion conformers/isomers of hexapeptide WHWLQL in the gas phase. Distinct H/D exchange trends for protonated WHWLQL ([M + H](+)) and its b(5)(+) and b(4)(+) fragment ions (with ND(3)) were observed. Isolated (12)C(all) isotopomers of both b(5)(+) and b(4)(+) fragment ions yielded bimodal distributions of H/D exchanged product ions. The H/D exchange reaction kinetics also confirmed that b(5)(+) and b(4)(+) fragment ions exist as combination of slow-exchanging ("s") and fast-exchanging ("f") species. The calculated rate constant for the first labile hydrogen exchange of [M + H](+) (k([M + H](+)) = 3.80 +/- 0.7 x 10(-10) cm(3) mol(-1) s( 1)) was approximately 30 and approximately 5 times greater than those for the "s" and "f" species of b(5)(+), respectively. Data from H/D exchange of isolated "s" species at longer ND(3) reaction times confirmed the existence of different conformers or isomers for b(5)(+) fragment ions. The sustained off-resonance irradiation collision-activated dissociation (SORI-CAD) of WHWLQL combined with the H/D exchange reactions indicate that "s" and "f" species of b(5)(+) and b(4)(+) fragment ions can be produced in the ICR cell as well as the ESI source. The significance of these observations for detailed understanding of protein sequencing and ion fragmentation pathways is discussed. PMID- 20005741 TI - Pulmonary artery catheter use with left-sided active endocarditis: deal or no deal? PMID- 20005742 TI - Cardiac football or cause of severe tricuspid regurgitation? PMID- 20005743 TI - Congenital pleuropericardial fistula complicating awake cardiac surgery. PMID- 20005745 TI - Con: general anesthesiologists should not be trained and certified in basic transesophageal echocardiography. PMID- 20005744 TI - Different anesthetic techniques associated with different incidences of chronic post-thoracotomy pain: low-dose remifentanil plus presurgical epidural analgesia is preferable to high-dose remifentanil with postsurgical epidural analgesia. AB - OBJECTIVE: To investigate the relationships between 2 anesthetic techniques, or the extent of allodynia around the surgical wound, and the occurrence of chronic post-thoracotomy pain. DESIGN: Prospective, randomized study. SETTING: A single institution, university hospital. PARTICIPANTS: Thirty-eight patients who underwent elective thoracotomy under general anesthesia. INTERVENTIONS: High-dose remifentanil (average effect-site concentration 5.61 +/- 0.84 ng/mL) with epidural analgesia started and at the end of surgery or low-dose remifentanil (average effect site concentration 1.99 +/- 0.02 ng/mL) with epidural analgesia with 0.5% ropivacaine started at the beginning of anesthesia. MEASUREMENTS AND MAIN RESULTS: Pain intensity and the extent of allodynia around the wound were measured during the hospital stay. The presence and intensity of residual pain were assessed 1, 3, and 6 months after surgery and at the end of the study (6-13 months, average 9 months). A DN4 neuropathic pain diagnostic questionnaire was conducted at the same times. In the high-dose group, the area with allodynia was three times larger than the area in the low-dose group. The increased allodynia was associated with a higher incidence of chronic pain (RR: 2.7-4.2) 3 and 6 months after surgery and at the end of the study (median follow-up: 9.5 months). CONCLUSIONS: High-dose remifentanil (0.14-0.26 microg/kg/min) without epidural analgesia during surgery is associated with a large area of allodynia around the wound. These patients develop a much higher incidence of chronic pain than those receiving low-dose remifentanil with epidural analgesia during surgery. PMID- 20005746 TI - Morphological analysis of pancreatic cystic masses. AB - RATIONALE AND OBJECTIVES: The aim of this study was to analyze the morphology of pancreatic cystic masses detected on multi-detector row computed tomography (MDCT) to determine whether single-dimension measurements of these masses are accurate reflections of their volumes. MATERIALS AND METHODS: Twenty-five pancreatic cystic masses detected on MDCT in 25 patients were evaluated. Pancreatic cysts were segmented on MDCT using commercially available software. All measurements were obtained twice by two independent investigators, and the means of values for segmented cyst volume (Vs) (milliliters), maximum transaxial diameter (millimeters), and elongation value (defined as 1 - [width/length], where 1 = ellipsoid and 0 = spherical) were reported for each cystic lesion. The volume of each cyst was also calculated (Vc) using the maximum transaxial diameter, with the hypothesis that the cyst was spherical. Student's t test was used to analyze the differences between values of Vs and Vc. Bland-Altman plots and Lin's concordance correlation were used to assess agreement between different measurement techniques. A P value < .05 denoted statistical significance. Interobserver variability was also determined using the Bland-Altman method. RESULTS: There was a significant difference between Vs and Vc (P < .0001). The elongation values ranged from 0.28 to 0.83 (mean, 0.57 +/- 0.12; median, 0.56). Mean interobserver variability was 1.7% (95% confidence interval, -4.89% to 8.30%). CONCLUSIONS: The results suggest that pancreatic cystic masses are not spherical. Therefore, a cyst's single largest transaxial dimension is not an accurate surrogate of its actual volume. PMID- 20005747 TI - Interest in conflicts anyone? PMID- 20005748 TI - The presence of circulating tumor cells does not predict extravesical disease in bladder cancer patients prior to radical cystectomy. AB - OBJECTIVE: Due to imprecise clinical staging, the finding of extravesical and node-positive disease at the time of radical cystectomy (RC) for patients with clinically localized bladder cancer is not uncommon. Circulating tumor cells (CTCs) have been shown to be present in the peripheral blood of patients with metastatic urothelial carcinoma. The object of this study was to evaluate the ability of CTCs to predict extravesical disease in bladder cancer patients prior to RC. MATERIALS AND METHODS: Peripheral blood samples from 43 patients with bladder cancer were evaluated using the CellSearch (Veridex, LLC, Raritan, NJ) CTC assay prior to RC. The sensitivity, specificity, and positive predictive value (PPV) of CTC status in predicting extravesical disease was calculated. Receiver operating characteristic (ROC) curves were generated to quantify the ability of CTCs to predict extravesical and node-positive disease. RESULTS: CTCs were detected in 9 (21%) patients prior to RC. The sensitivity, specificity, and PPV of CTC status in predicting extravesical disease were 27%, 88% and 78%, respectively. The accuracy of CTC status in predicting extravesical (>=pT3 or node-positive) disease for the entire cohort was 0.576. In a model incorporating preoperative hydronephrosis, CTC status did not improve the predictive accuracy for extravesical disease (0.576 vs. 0.585, P = 0.915). CONCLUSION: CTCs were detected in low numbers in a small percentage (21%) of patients prior to undergoing RC at our institution. CTC status was not a robust predictor of extravesical or node-positive disease in this cohort. CTC status is not likely to be a clinically useful parameter for directing therapeutic decisions in patients with <=cT2 bladder cancer. PMID- 20005749 TI - Acute toxicity of image-guided hypofractionated radiotherapy for prostate cancer: nonrandomized comparison with conventional fractionation. AB - OBJECTIVES: To compare acute toxicity of prostate cancer image-guided hypofractionated radiotherapy (hypo-IGRT) with conventional fractionation without image-guidance (non-IGRT). To test the hypothesis that the potentially injurious effect of hypofractionation can be counterbalanced by the reduced irradiated normal tissue volume using IGRT approach. MATERIALS AND METHODS: One hundred seventy-nine cT1-T2N0M0 prostate cancer patients were treated within the prospective study with 70.2 Gy/26 fractions (equivalent to 84 Gy/42 fractions, alpha/beta 1.5 Gy) using IGRT (transabdominal ultrasound, ExacTrac X-Ray system, or cone-beam computer tomography). Their prospectively collected data were compared with data of 174 patients treated to 80 Gy/40 fractions with non-IGRT. The difference between hypo-IGRT and non-IGRT cohorts included fractionation (hypofractionation vs. conventional fractionation), margins (hypo-IGRT margins: 7 mm and 3 mm, for all but posterior margins; respectively; non-IGRT margins: 10 and 5 mm, for all but posterior margins, respectively), and use of image-guidance or not. Multivariate analysis was performed to define the tumor-, patient-, and treatment-related predictors for acute toxicity. RESULTS: All patients completed the prescribed radiotherapy course. Acute toxicity in the hypo-IGRT cohort included rectal (G1: 29.1%; G2: 11.2%; G3: 1.1%) and urinary events (G1: 33.5%; G2: 39.1%; G3: 5%). Acute toxicity in the non-IGRT patients included rectal (G1: 16.1%; G2: 6.3%) and urinary events (G1: 36.2%; G2: 20.7%; G3: 0.6%). In 1 hypo IGRT and 2 non-IGRT patients, radiotherapy was temporarily interrupted due to acute toxicity. The incidence of mild (G1-2) rectal and bladder complications was significantly higher for hypo-IGRT (P = 0.0014 and P < 0.0001, respectively). Multivariate analysis showed that hypo-IGRT (P = 0.001) and higher PSA (P = 0.046) are correlated with higher acute urinary toxicity. No independent factor was identified for acute rectal toxicity. No significant impact of IGRT system on acute toxicity was observed. CONCLUSIONS: The acute toxicity rates were low and similar in both study groups with some increase in mild acute urinary injury in the hypo-IGRT patients (most probably due to the under-reporting in the retrospectively analyzed non-IGRT cohort). The higher incidence of acute bowel reactions observed in hypo-IGRT group was not significant in the multivariate analysis. Further investigation is warranted in order to exclude the bias due to the nonrandomized character of the study. PMID- 20005751 TI - Peak wall stress does not necessarily predict the location of rupture in abdominal aortic aneurysms. AB - Using finite element analysis, we evaluated if the site of an aortic bleb, known to be prone to rupture, coincides with the location of peak wall stress (PWS) in a patient-specific abdominal aortic aneurysm (AAA) model. REPORT: PWS was not located at the bleb site, even when stress values were estimated for different bleb wall thicknesses (0.5-2.0 mm) while the rest of the AAA wall was considered constant (2 mm). DISCUSSION: The sites of PWS in AAAs should not always be considered as the sites most prone to rupture since other factors, such as wall strength, may play a role in rupture-risk prediction, depicting the need for further investigation of these parameters. PMID- 20005750 TI - Systematic preoperative coronary angiography and stenting improves postoperative results of carotid endarterectomy in patients with asymptomatic coronary artery disease: a randomised controlled trial. AB - OBJECTIVE: To evaluate the usefulness of systematic coronary angiography followed, if needed, by coronary artery angioplasty (percutaneous coronary intervention (PCI)) on the incidence of cardiac ischaemic events after carotid endarterectomy (CEA) in patients without evidence of coronary artery disease (CAD). MATERIALS AND METHODS: From January 2005 to December 2008, 426 patients, candidates for CEA, with no history of CAD and with normal cardiac ultrasound and electrocardiography (ECG), were randomised into two groups. In group A (n=216) all the patients had coronary angiography performed before CEA. In group B, all the patients had CEA without previous coronary angiography. In group A, 66 patients presenting significant coronary artery lesions at angiography received PCI before CEA. They subsequently underwent surgery under aspirin (100 mg day( 1)) and clopidogrel (75 mg day(-1)). CEA was performed within a median delay of 4 days after PCI (range: 1-8 days). Risk factors, indications for CEA and surgical techniques were comparable in both groups (p>0.05). The primary combined endpoint of the study was the incidence of postoperative myocardial ischaemic events combined with the incidence of complications of coronary angiography. Secondary endpoints were death and stroke rates after CEA and incidence of cervical haematoma. RESULTS: Postoperative mortality was 0% in group A and 0.9% in group B (p=0.24). One postoperative stroke (0.5%) occurred in group A, and two (0.9%) in group B (p=0.62). No postoperative myocardial event was observed in group A, whereas nine ischaemic events were observed in group B, including one fatal myocardial infarction (p=0.01). Binary logistic regression analysis demonstrated that preoperative coronary angiography was the only independent variable that predicted the occurrence of postoperative coronary ischaemia after CEA. The odds ratio for coronary angiography (group A) indicated that when holding all other variables constant, a patient having preoperative coronary angiography before carotid surgery was 4 times less likely to have a cardiac ischaemic event after carotid surgery. No complications related to coronary angiography were observed and no cervical haematomas occurred in patients undergoing surgery under aspirin and clopidogrel in this study. CONCLUSIONS: Systematic preoperative coronary angiography, possibly followed by PCI, significantly reduces the incidence of postoperative myocardial events after CEA in patients without clinical evidence of CAD. PMID- 20005752 TI - Correction of square jaw with low angles using mandibular "V-line" ostectomy combined with outer cortex ostectomy. AB - OBJECTIVE: The objective of this study was to evaluate the feasibility and effectiveness of correction for square jaw with low angle using mandibular "V line" ostectomy combined with outer cortex ostectomy. STUDY DESIGN: From July 2005 to November 2007, 31 patients who had square faces with low angles were treated with mandibular "V-line" ostectomy combined with mandibular outer cortex ostectomy. All the patients had standard frontal and lateral cephalometric radiographs, panoramic radiographs, and were photographed preoperatively and postoperatively to assess their face contour. The alteration of mandibular angle and width of lower face was observed for 6 to 24 months postoperatively. RESULTS: Postoperative appearance of all 31 cases showed that the width of lower face was reduced expectedly; the gonial angle and the mandibular plane angle were increased effectively. The final aesthetic outcomes were quite satisfactory in all cases for both the surgeons and the patients. CONCLUSION: A slender oval facial outline as the fashionable aesthetics could be achieved by using mandibular "V-line" ostectomy combined with mandibular outer cortex ostectomy. It might be a feasible and a reasonable surgical procedure for correction of square jaw with low angle. PMID- 20005753 TI - Under diagnosis of canine cognitive dysfunction: a cross-sectional survey of older companion dogs. AB - Canine cognitive dysfunction (CCD) is a neurobehavioural syndrome affecting aged dogs. Using a large cross-sectional epidemiological study of older dogs, this study aimed to estimate the prevalence of CCD amongst community based dogs (mean age 11.67years; range 8-19.75) and to determine the rate of veterinary diagnosis amongst affected dogs. An 84-item questionnaire was used to obtain information across six behavioural domains. Of the eligible survey responses obtained (n=957) a randomly selected one-half (n=497) was used for this study. Using a provisional diagnosis based on 27 significant behavioural items, the prevalence rate of CCD was estimated to be 14.2%. This was in contrast with only 1.9% diagnosed with CCD by a veterinarian. There was an exponential increase in prevalence of CCD with age (R2=0.9435), but prevalence did not differ by breed size or between longevity groups. The prevalence rate of CCD reported here is consistent with previous findings, and further supports the contention that the majority of these dogs do not receive a formal diagnosis. PMID- 20005755 TI - Application of z-COSY experiment and its variant for accurate chiral discrimination by (1)H NMR. AB - We report the application of z-COSY experiment and a band selected version of it by employing a selective 90 degrees pulse entitled BASE-z-COSY for precise chiral discrimination, quantification of enantiomeric excess and the analyses of the (1)H NMR spectra of chiral molecules aligned in the chiral liquid crystalline solvent poly-gamma-benzyl-l-glutamate (PBLG). We have demonstrated their applicability for obtaining very high resolution in the (1)H NMR spectra of small organic molecules. It is well known that the commonly employed z-COSY experiment disentangles the spectral complexity, provides pure phase spectra with high resolution, aids in the complete spectral analyses, in addition to yielding information on relative signs of the couplings. The BASE-z-COSY experiment possesses all these properties, permits the measure of enantiomeric excess, in addition to large saving of instrument time. PMID- 20005754 TI - Chronic impact of topiramate on acid-base balance and potassium in childhood. AB - Topiramate, which is commonly prescribed for seizure disorders and migraine prophylaxis, sometimes causes metabolic acidosis and hypokalemia. Since the effects of topiramate on acid-base balance and potassium levels have not been well explored in children, acid-base balance, anion gap and potassium were assessed in 24 patients (8 females and 16 males) aged between 4.6 and 19 years on topiramate for more than 12 months and in an age-matched control group. Plasma bicarbonate (21.7 versus 23.4 mmol/L; P<0.03), carbon dioxide pressure (39.7 versus 43.2mm Hg; P<0.05), and potassium (3.7 versus 4.0 mmol/L; P<0.03) were on the average lower and chloride (109 versus 107 mmol/L; P<0.03) higher in patients treated with topiramate than in controls. Blood pH, plasma sodium and the anion gap were similar in patients on topiramate and in controls. In patients on topiramate no significant correlation was observed between the dosage of this agent and plasma bicarbonate or potassium as well as between topiramate blood level and the mentioned electrolytes. In conclusion long-term topiramate treatment is associated with a mild, statistically significant tendency towards compensated normal anion gap metabolic acidosis and hypokalemia. PMID- 20005756 TI - Combined pharmacophore and structure-guided studies to identify diverse HSP90 inhibitors. AB - Heat Shock Protein 90 (HSP90), an ATP-dependent molecular chaperone, has emerged as a promising target in the treatment of cancer. Inhibition of HSP90 represents a new target of antitumor therapy, since it may influence many specific signaling pathways. Many HSP90 inhibitors bind to the ATP-binding pocket, inhibit chaperone function, resulting in cell death. Recent clinical trials for treatment of cancer have put HSP90's importance into focus and have highlighted the need for full scale research into HSP90 related pathways. Here we report five novel HSP90 inhibitors which were identified by using pharmacophore models and docking studies. We used highly discriminative pharmacophore model as a 3D query to search against database of approximately 1 M compounds and cluster analysis results yielded 455 compounds which were further subjected for docking. Glide docking studies suggested 122 compounds as in silico hits and these compounds were further selected for the cytotoxicity assay in the HSP90-over expressing SKBr3 cell line. Of the 122 compounds tested, 5 compounds inhibited cell growth with an IC(50) value less than 50 microM. PMID- 20005757 TI - A novel PCFT gene mutation (p.Cys66LeufsX99) causing hereditary folate malabsorption. AB - Hereditary folate malabsorption (HFM) is a rare autosomal recessive disorder which is characterized by impaired intestinal folate malabsorption and impaired folate transport into the central nervous system. Mutations in the intestinal folate transporter PCFT have been reported previously in only 10 individuals with this disorder. The purpose of the current study was to describe the clinical phenotype and determine the molecular basis for this disorder in a family with four affected individuals. A consanguineous family of Pakistani origin with autosomal recessive HFM was ascertained and clinically phenotyped. After genetic linkage studies all coding exons of the PCFT gene were screened for mutations by direct sequencing. The clinical phenotype of four affected patients is described. Direct sequencing of PCFT revealed a novel homozygous frameshift mutation (c.194dupG) at a mononucleotide repeat in exon 1 predicted to result in a truncated protein (p.Cys66LeufsX99). This report extends current knowledge on the phenotypic manifestations of HFM and the PCFT mutation spectrum. PMID- 20005758 TI - Effect of water source on intake and urine concentration in healthy cats. AB - Increasing water intake and decreasing urine concentration are recommended for cats with urolithiasis and with idiopathic cystitis. Fountains are advocated to encourage drinking; however, effects on drinking of fountains have not been reported in cats living in pet owners homes. Thirteen healthy cats were assigned to have 24-h water intake and urine osmolality and specific gravity measured when water was offered from a bowl or fountain. One cat developed excessive barbering, vomiting, and refusal to drink water offered from the fountain. For the remaining 12 cats, intake was slightly greater from the fountain. However, urine osmolality was not significantly different. In this study, a fountain failed to substantially increase water intake and dilute urine in cats. A similar study including a greater period of time and additional cats may clarify the results of this study. PMID- 20005759 TI - Tuberculous meningitis in HIV-infected patients in Brazil: clinical and laboratory characteristics and factors associated with mortality. AB - BACKGROUND: Tuberculous meningitis (TBM) is a growing problem in HIV-infected patients in developing countries, where there is scarce data about this co infection. Our objectives were to analyze the main features and outcomes of HIV infected patients with TBM. METHODS: This was a retrospective study of HIV infected Brazilian patients admitted consecutively for TBM. All patients had Mycobacterium tuberculosis isolated from the cerebrospinal fluid (CSF). Presenting clinical and laboratory features were studied. Multivariate analysis was used to identify variables associated with death during hospitalization and at 9 months after diagnosis. Survival was estimated using the Kaplan-Meier method. RESULTS: We included 108 cases (median age 36 years, 72% male). Only 15% had fever, headache, and meningeal signs simultaneously. Forty-eight percent had extrameningeal tuberculosis. The median CD4+ cell count was 65 cells/microl. Among 90 cases, 7% had primary resistance to isoniazid and 9% presented multidrug resistant strains. The overall mortality during hospitalization was 29% and at 9 months was 41%. Tachycardia and prior highly active antiretroviral therapy (HAART) were associated with 9-month mortality. The 9-month survival rate was 22% (95% confidence interval 12-43%). CONCLUSIONS: Clinical and laboratory manifestations were unspecific. Disseminated tuberculosis and severe immunosuppression were common. Mortality was high and the 9-month survival rate was low. Tachycardia and prior HAART were associated with death within 9 months of diagnosis. PMID- 20005760 TI - A newly identified Crimean-Congo hemorrhagic fever virus strain in Turkey. AB - Crimean-Congo hemorrhagic fever (CCHF) is a fatal viral disease that occurs in approximately 30 countries. It has the most extensive geographic range among the tick-borne viruses that affect human health. Recently, a 6-year-old boy presented with complaints of fever, fatigue, and loss of appetite. He revealed a history of tick bite in rural Istanbul three days prior to presentation. A hyperemia was detected at the site of the tick bite. Laboratory tests showed that alanine aminotransferase, aspartate aminotransferase, lactate dehydrogenase, and creatine phosphokinase levels were elevated and that the prothrombin time and activated partial thromboplastin time were prolonged. Anti-CCHF virus IgM ELISA and a reverse transcriptase-PCR assay for CCHF RNA were both positive. Phylogenetic studies revealed that the virus was a new AP92-like CCHF strain, which was named KMAG-Hu-07-01 (accession number EU057975). This patient could provide important information on the transmission dynamics of CCHF infection. PMID- 20005762 TI - The efficacy of catheters coated with minocycline and rifampin in the prevention of catheter-related bacteremia in cancer patients receiving high-dose interleukin 2. AB - High-dose interleukin-2 (HDIL-2) has proven to be an effective treatment for metastatic renal cell carcinoma and melanoma. Previous studies have shown an increase in catheter-related bacteremia (CRB) in patients on HDIL-2. The primary objective of this study was to evaluate the effectiveness of minocycline and rifampin-coated catheters (M/R-C) in reducing CRB in cancer patients on HDIL-2. This was a retrospective study where non-coated catheters (NC-C) and M/R-C were used for the administration of HDIL-2 before and after December 2004, respectively. Data collected included demographics, cancer type, catheter type, antibiotic prophylaxis, and infection rates. A total of 107 episodes of catheter use for HDIL-2 were evaluated in 78 patients (30 episodes in patients with M/R-C vs. 77 with NC-C). A total of nine episodes of CRB were identified, all in patients with NC-C (M/R-C 0% vs. NC-C 12%; p=0.06). The median time to bacteremia was 11 days (range 1-315 days). A log-rank test showed a trend that the M/R-C group had lower probability of getting CRB than the NC-C group (p=0.06). The use of M/R-C in patients on HDIL-2 therapy for advanced melanoma and renal cell carcinoma may have reduced the risk of CRB to nil. CRB still occurred despite antibiotic prophylaxis in patients with NC-C. PMID- 20005763 TI - Treatment of vancomycin-intermediate Staphylococcus aureus (VISA) endocarditis with linezolid. AB - We report a case of infective endocarditis due to vancomycin-intermediate Staphylococcus aureus (VISA). This was treated with a combination of intravenous linezolid and fusidic acid. Cure was achieved without surgical intervention. PMID- 20005764 TI - [Spinals and paraspinals tumors treated by CyberKnife: feasibility and efficacy]. AB - PURPOSE: Stereotactic radiotherapy using the CyberKnife has become a key treatment in the multidisciplinary management of secondary tumours, as well as primary benign or malignant tumours located within or adjacent to vertebral bodies and the spinal cord. The aim of this treatment is to improve local control and clinical response, including previously irradiated cases. PATIENTS AND METHODS: In this study, we present the first patients treated with CyberKnife between December 2006 and December 2007 for spinal or paraspinal tumours. The primary aim was to assess the feasibility and tolerance of stereotactic radiotherapy using the CyberKnife. Secondary aims were to establish the short term local control, to calculate the local progression-free survival and overall survival. Clinical examination and imaging procedures were performed every three months. Response was assessed according to RECIST criteria. RESULTS: During that period, 16 patients were treated with CyberKnife. Thirteen patients had been pre treated, three of whom had received spinal cord doses considered to be maximal. Three patients did not receive previous irradiation. The median age was 59 (36 74). The most frequent symptoms were pain (n = 8) and motor weakness (n = 4). The median dose was 30 Gy (16-50). The median number of fractions was 3 (1-5). No patient developed acute myelitis. Three patients developed acute reaction. Overall survival at 18 months was 72.4%, with a mean survival of 18.2 months (95% CI: 15.4-20.9). Local progression-free survival at 18 months was 58.4%, with a mean value of 16.9 months (95% CI: 13.6-20.2). CONCLUSION: The use of stereotactic radiotherapy with CyberKnife represents a major progress in the management of paraspinal tumours. The main advantages are better sparing of the spinal cord and the possibility of increasing the dose to the tumour target volume. PMID- 20005765 TI - [Feasibility and toxicity of a single fraction high-dose-rate brachytherapy followed by a course of EBRT for localized prostate cancer: a retrospective study. The Polyclinique Courlancy experience]. AB - PURPOSE: Evaluate the feasibility and toxicity of radiation dose escalation delivered with a single fraction high-dose-rate (HDR) brachytherapy boost followed by external beam radiotherapy for intermediate and high risk localized prostate cancer - a retrospective study. PATIENTS AND METHODS: Between December 2004 and December 2008, 61 patients with intermediate risk or high-risk localized prostate cancer received a single 10 Gy fraction of interstitial HDR brachytherapy followed by a 64 Gy course of external beam radiation therapy. Dose volume histograms, conformity index and side effects were systematically analyzed. RESULTS: HDR brachytherapy dosimetric criteria were respected. Early side effects (< or = 3 months after full treatment): 30 % reported grade 2 or grade 3 urinary toxicity and 26 % reported grade 2 or grade 3 bowel toxicity were reported. Late side effects (> 3 months): 12 % reported grade 2 or grade 3 urinary toxicity and 5 % reported grade 2 or grade 3 bowel toxicity were reported. No patients reported any grade 4 late toxicity events. Three months after treatment, 7 % grade 1, 25 % grade 2 and 39 % grade 3 erectile dysfunction were reported. CONCLUSION: Our monofractionation protocol is an easy technique to implement logistically. Acute and late toxicities are acceptable and comparable to those published by various teams mostly using multifractionation protocols. A longer follow-up is required to assess the effect of this dose escalation protocol on long-term biological control. PMID- 20005766 TI - Sensitivity and specificity of the finger tapping task for the detection of psychogenic movement disorders. AB - Psychogenic movement disorders (PMD) represent a diagnostically challenging group of patients in movement disorders. Finger tapping tests (FTT) have been used in neuropsychiatric evaluations to identify psychogenic conditions, but their use in movement disorders has been limited to the quantification of upper extremity disability in idiopathic Parkinson disease (IPD). We evaluated the ability of the FTT to objectively identify PMD by screening 195 individuals from a movement disorder clinic with IPD, dystonia, essential tremor, or PMD and compared them to 130 normal adults. All subjects performed six-30 s trials using alternate hands. We compared mean FTT score and the coefficient of variation between diagnostic groups. FTT scores in IPD were inversely correlated with Hoehn and Yahr stage (p < 0.001) and the United Parkinson Disease Rating Scale III (motor) subscale (p < 0.001). FTT scores were significantly lower in PMD (mean = 41.72) when compared to the other diagnostic groups after controlling for age. The coefficient of variation was not significantly different between diagnostic groups. ROC analysis identified a cutoff FTT ratio of 0.670 or less was 89.1% specific and 76.9% sensitive for the diagnosis of PMD. We conclude the FTT can provide supportive evidence for the diagnosis of PMD. PMID- 20005767 TI - The coordination of movement: optimal feedback control and beyond. AB - Optimal control theory and its more recent extension, optimal feedback control theory, provide valuable insights into the flexible and task-dependent control of movements. Here, we focus on the problem of coordination, defined as movements that involve multiple effectors (muscles, joints or limbs). Optimal control theory makes quantitative predictions concerning the distribution of work across multiple effectors. Optimal feedback control theory further predicts variation in feedback control with changes in task demands and the correlation structure between different effectors. We highlight two crucial areas of research, hierarchical control and the problem of movement initiation, that need to be developed for an optimal feedback control theory framework to characterise movement coordination more fully and to serve as a basis for studying the neural mechanisms involved in voluntary motor control. PMID- 20005768 TI - Overexpression of Kif2a promotes the progression and metastasis of squamous cell carcinoma of the oral tongue. AB - The aim of this study was to examine the Kif2a expression and its role in tumor progression, invasion and metastasis in squamous cell carcinoma of the oral tongue (SCCOT). The study included 44 cases of primary tumor and the corresponding adjacent tissues, 20 cases of primary tumor with lymph node metastasis. Immunohistochemistry was used to observe the Kif2a expression and its correlation with clinicopathologic factors in oral tongue cancer. The immunohistochemistry showed that Kif2a expression was stronger in oral tongue cancer tissues than in paired adjacent tissues (P<0.01), and the higher expression of Kif2a was also significantly associated with lymph node metastasis (P<0.01), tumor clinical stage (P<0.01). In addition, in vitro results from transwell chamber assay showed that Tca8113 cells transfected with Kif2a-siRNA had a decreased migratory ability (P<0.01) compared to nonsense-siRNA-transfected cells. Therefore we speculate the overexpression of Kif2a might be involved in the progression, invasion and metastasis of SCCOT and Kif2a should be as a predictor for prognosis. PMID- 20005769 TI - Predictors of locoregional recurrence in early stage oral cavity cancer with free surgical margins. AB - Locoregional recurrence in patients with early stage oral cavity squamous cell carcinoma (ESOSCC) after surgery remains a problem and can affect their survival. We sought to identify new high-risk factors in these patients, who need further adjuvant therapy. We retrospectively reviewed records for 148 patients who underwent surgery for ESOSCC between 2002 and 2006 with negative surgical margins. The primary endpoint was locoregional recurrence. Recurrence-free survival (RFS) and overall survival (OS) were calculated by the Kaplan-Meier method. Univariate and multivariate analyses were used to identify independent predictors of locoregional recurrence. All patients were grouped into the low- and high-risk groups according to the odds ratios (OR) of the predictors. Recurrence rates of the low- and high-risk groups were then predicted. Recurrence was observed in 17 of 148 (11.5%) patients at the end of this study. None of the patients received postoperative radiotherapy or chemotherapy. At 3 years, the RFS rate was 89.7% and the OS rate at 3 years was 84.1%. Univariate analysis of the RFS revealed three significant prognostic factors: lymphovascular permeation (LVP, p<0.001), perineural infiltration (PNI, p=0.08), and non-T4 muscular invasion (non-T4MI, p<0.005). Multivariate analysis demonstrated that LVP (p=0.007, OR=10.7) and non-T4 MI (p=0.001, OR=8.347) were independent predictors. The recurrence rate was 1.96% in patients without LVP or non-T4MI, and it increased to 26.47% in patients with non-T4MI, to 50% in patients with LVP, and to 50% in patients with both. According to the status of LVP and non-T4MI, patients were divided into two groups: low-risk (no factors present) and high risk (one or both factors present) groups. The 2-year RFS was lower in the high risk group (84.13%) than in the low-risk group (93.91%); the 3-year RFS was also lower in the high-risk group (70.49%) than in the low-risk group (91.99%) (p=0.008). Subgroup analysis revealed that elective neck dissections did not affect the outcome or change the pattern of failure. For patients with elective neck dissections, the RFS was lower in the high-risk group than in the low-risk group (p=0.03). In ESOSCC (pT1-2N0), LVP and non-T4MI significantly increased the recurrence rate. The presence of one or both factors (LVP and/or non-T4MI) should be considered as a high-risk condition for locoregional recurrence, and adjuvant therapy is needed in such cases. PMID- 20005770 TI - Enhanced frequency upconversion of Sm3+ ions by elliptical Au nanoparticles in dichroic Sm3+: Au-antimony glass nanocomposites. AB - Dichroic Sm(3+): Au-antimony glass nanocomposites are synthesized in a new reducing glass (dielectric) matrix (mol%) K(2)O-B(2)O(3)-Sb(2)O(3) (KBS) by a single-step melt-quench technique involving selective thermochemical reduction. X ray diffraction (XRD) and selected area electron diffraction (SAED) results indicate that Au(0) nanoparticles are grown along the (200) plane direction. The transmission electron microscopic (TEM) image reveals the elliptical Au(0) nanoparticles having major axis range 12-17 nm. Dichroic behavior is due to elliptical shape of Au(0) nanoparticles of aspect ratio approximately 1.2. Au(0) NPs of concentration of 0.03 wt% (4.1 x 10(18)atoms/cm(3)) drastically enhances the intensity ( approximately 7-folds) of electric dipole (4)G(5/2)-->(6)H(9/2) red transition (636 nm) of Sm(3+) ions and then attenuates with further increase in Au(0) concentration. The magnetic dipole (4)G(5/2)-->(6)H(5/2) green (566 nm) and (4)G(5/2)-->(6)H(7/2) orange (602 nm) transitions remain almost unaffected by presence of nano Au(0). Local field enhancement (LFE) induced by Au(0) SPR and energy transfer (ET) from fluorescent Au(0)-->Sm(3+) ions are found to be responsible for the enhancement while reverse ET from Sm(3+)-->Au(0) and optical re-absorption due to Au(0) SPR for attenuation. PMID- 20005771 TI - Challenges to inter-departmental coordination of patient transfers: a workflow perspective. AB - OBJECTIVE: The purpose of this study is to identify challenges to inter departmental coordination activities that affect patient transfer workflow and to provide socio-technical requirements for the design of technologies to better support patient transfer workflow. DESIGN: We conducted our study in two clinical and one non-clinical department at a major academic hospital. We utilized qualitative data collection techniques including observations of patient transfer practices of the different departments and interviews with departmental staff to collect data on the inter-departmental coordination activities and its effect on patient transfer workflow. RESULTS: We identified three inter-departmental challenges that affected the patient transfer workflow: ineffective inter departmental interactions, ineffective information handoffs, and ineffectiveness of current information technologies. DISCUSSION: To address these challenges, we discuss three socio-technical design requirements that designers need to pay attention to while developing inter-departmental healthcare information systems. To ensure effective inter-departmental coordination, the systems should incorporate features that can support the mediating role of integrators, the collaborative balancing of goals, and the collaborative prioritization of resources. PMID- 20005772 TI - Do we need to consider non-linear information flow in corticomuscular interaction? PMID- 20005773 TI - REM sleep characteristics of nightmare sufferers before and after REM sleep deprivation. AB - OBJECTIVES: To examine whether disrupted regulation of REM sleep propensity is implicated in nightmare (NM) pathophysiology. BACKGROUND: Heightened REM propensity induced by REM sleep deprivation is belied by increases in REM %, REM density and the dream-like quality of dream mentation during post-deprivation recovery sleep. Compromised regulation of REM sleep propensity may be a contributing factor in the pathophysiology of frequent NMs. METHODS: A preliminary study of 14 subjects with frequent NMs (> or = 1 NM/week; 27.6+/-9.9 years) and 11 healthy control subjects (<1 NM/month; 24.3+/-5.3 years) was undertaken. Subjects completed home sleep/dream logs and underwent three nights of polysomnographic recording with REM sleep deprivation on night 2. Group differences were assessed for a battery of REM sleep and dream measures on nights 1 and 3. RESULTS: Several measures, including #skipped early-night REM periods, REM latency, REM/NREM cycle length, early/late REM density, REM rebound, late night REM% and dream vividness, suggested that REM sleep propensity was abnormally low for the frequent NM group throughout the 3-day study. CONCLUSIONS: Findings raise the possibility that REM anomalies recorded from NM sufferers sleeping in the laboratory environment reflect a disruption of one or more endogenous regulators of REM sleep propensity. PMID- 20005774 TI - Relationships among dietary nutrients and subjective sleep, objective sleep, and napping in women. AB - OBJECTIVE: To describe which dietary nutrient variables are related to subjective and objective habitual sleep and subjective and objective napping. METHODS: Participants were 459 post-menopausal women enrolled in the Women's Health Initiative. Objective sleep was estimated using one week of actigraphy. Subjective sleep was prospectively estimated with a daily sleep diary. Dietary nutrients were calculated from food frequency questionnaires. RESULTS: The most significant correlations were with subjective napping, including (from strongest to weakest): total fat, calories, saturated fat, monounsaturated fat, trans fat, water, proline, serine, tyrosine, phenylalanine, valine, cholesterol, leucine, glutamic acid, ash, isoleucine, histidine, sodium, tryptophan, protein, threonine, cystine, methionine, phosphorous, polyunsaturated fat, animal protein, aspartic acid, arginine, lysine, alanine, caffeine, riboflavin, gamma-tocopherol, glycine, retinol, delta-tocopherol, Vitamin D, and selenium. Actigraphic nocturnal sleep duration was negatively associated with total fat, monounsaturated fat, trans fat, saturated fat, polyunsaturated fat, calories, gamma-tocopherol, cholesterol, and alpha-tocopherol-eq. CONCLUSIONS: Actigraphic total sleep time was negatively associated with intake of fats. Subjective napping, which may be a proxy for subjective sleepiness, was significantly related to fat intake as well as intake of meat. PMID- 20005775 TI - New approaches for functional genomic studies in staphylococci. AB - Functional transcriptomics studies have resulted in interesting insights into Staphylococcus aureus diversity and pathogenicity. Here we review the principles, advantages and disadvantages of recent technical developments in the field of transcriptomics and their potential impact on S. aureus research. PMID- 20005776 TI - An oldie but a goodie - cell wall biosynthesis as antibiotic target pathway. AB - Bacterial cell wall biosynthesis represents the target pathway for penicillin, the first antibiotic that was clinically applied on a large scale. Penicillin, by means of its beta-lactam ring, inhibits a number of enzymes which participate in inserting monomeric cell wall building blocks into the cell wall polymer and which have been termed penicillin-binding proteins (PBPs). Ever since the introduction of penicillin, hundreds of beta-lactam antibiotics have been developed and details of their molecular activities elaborated. Meanwhile, various additional classes of antibiotics have been described, which inhibit the same pathway, yet use target molecules others than the PBPs. Such classes include the glycopeptide antibiotics, lipopeptide and lipodepsipeptide antibiotics, the lantibiotics and various other natural product antibiotics with comparatively complex structures. They usually target the membrane-bound steps of the biosynthesis pathway and the highly conserved lipid-bound intermediates of the building block such as lipid II, which represents a particular "Achilles' heel" for antibiotic attack. With in-depth analysis of the activity of more recently identified inhibitors and with the availability of novel techniques for studying prokaryotic cell biology, new insights were obtained into the molecular organisation of the cell wall biosynthesis machinery and its interconnections with other vital cellular processes such as cell division. This, in turn, provides hints for new targets to be exploited and for the development of novel cell wall biosynthesis inhibitors. PMID- 20005777 TI - Emergence of methicillin-resistant Staphylococcus aureus (MRSA) in different animal species. AB - The emergence of methicillin-resistant Staphylococcus aureus (MRSA) in animals such as horses, pet animals and productive livestock has raised questions of a probable human origin and in more general of host specificity of S. aureus. Particular clonal lineages are obviously specific for humans (e.g. ST15, ST25, ST45) and other for ruminants (e.g. ST151). MRSA associated with veterinary nosocomial infections (e.g. ST8 and ST254 in horses, ST22 in small animals) very likely have their origin in health care facilities. MRSA ST398 which became first known from widespread colonization in industrially raised pigs seems to have a limited host specificity and is able to colonize and to cause infections in various hosts. Mechanisms of host adaptation and their genomic background are poorly understood so far. PMID- 20005778 TI - Left main coronary artery atresia diagnosed with 128-MDCT. PMID- 20005779 TI - Massive haemothorax due to subclavian artery rupture: emergency thoracotomy or primary stent-grafting? PMID- 20005781 TI - Cytokine genes are associated with tuberculin skin test response in a native Brazilian population. AB - Tuberculosis was a major cause of population decline among Brazilian indigenous peoples and remains a leading cause of morbidity and mortality among them. Despite high BCG coverage, results of Tuberculin Skin Test (TST) reactivity have shown high rates of anergy in Amazonian Indians. Given the high prevalence of anergy in these populations and the fact that genetic host factors play an important role in susceptibility to Mycobacterium tuberculosis (MTB), the aim of this study was to evaluate the association of nineteen polymorphisms in fifteen genes related to immune response and anergy in the Xavante, an indigenous group from Brazil. A total of 481 individuals were investigated. TST anergy was observed in 69% of them. Polymorphisms in four genes showed absence or very low variability: SP110, PTPN22, IL12RB1 and IL6. IFNG +874 A/T heterozygotes and IL4 590 C/C homozygotes were more frequent in those individuals who presented a positive TST (prevalence ratios of 1.9 and 2.0 respectively). The risk of anergy was 1.5 in IL10-1082 G/G homozygotes when compared to carriers for the A allele. In indigenous groups such as the Xavante exposure to a variety of infections, associated with specific genetic factors, may disturb the T-helper 1 and T-helper 2 balance leading to increased immunological susceptibility. PMID- 20005782 TI - Magnesium in obstetrics. AB - Magnesium is a critical physiological ion, and magnesium deficiency might contribute to the development of pre-eclampsia, to impaired neonatal development and to metabolic problems extending into adult life. Pharmacologically, magnesium is a calcium antagonist with substantial vasodilator properties but without myocardial depression. Cardiac output usually increases following magnesium administration, compensating for the vasodilatation and minimising hypotension. Neurologically, the inhibition of calcium channels and antagonism of the N-methyl d-aspartic acid (NMDA) receptor raises the possibility of neuronal protection, and magnesium administration to women with premature labour may decrease the incidence of cerebral palsy. It is the first-line anticonvulsant for the management of pre-eclampsia and eclampsia, and it should be administered to all patients with severe pre-eclampsia or eclampsia. Magnesium is a moderate tocolytic but the evidence for its effectiveness remains disputed. The side effects of magnesium therapy are generally mild but the major hazard of magnesium therapy is neuromuscular weakness. PMID- 20005783 TI - Metabolic acuity score: effect on major complications after bariatric surgery. AB - BACKGROUND: Co-morbid conditions in obese patients contribute to the incidence and severity of major complications after bariatric surgery and significantly increase the cost of the procedure. Previous publications have validated the patient factors that increase the risk of mortality; however, it is currently a rare event. The development of a metabolic acuity score (MAS) to augment the body mass index might allow for accurate preoperative assessment and optimal treatment of patients. The present study has proposed a MAS for decreasing major complications. METHODS: Prospectively collected outcomes of 2416 patients undergoing Roux-en-Y gastric bypass (n = 1821) or laparoscopic adjustable gastric banding (n = 595) in a community hospital were evaluated for the incidence of major complications, readmissions, and reoperations. Beginning in August of 2006, 1072 patients were divided into MAS groups of 1-4 according to age, body mass index, weight, history of deep vein thrombosis/pulmonary embolism, sleep apnea, diabetes, hypertension, immobility, heart disease, and psychological classification. The acuity groups were compared with each other and with 1344 patients who underwent treatment before the MAS was implemented. RESULTS: A significant decrease occurred in the readmission rates within 30 days after the MAS was put into practice (8.5% before MAS versus 1.7% after MAS, P <.001) for the Roux-en-Y gastric bypass patients. The postoperative infection rates were lower after implementing the MAS (3.5% before MAS, .7% after MAS, P <.001). After adjusting for random and fixed effects of covariates, the implementation of the MAS significantly reduced the incidence of postoperative internal hernias, infections, deep vein thrombosis, readmissions, and reoperations. CONCLUSION: Recognition of specific patient acuity characteristics through the implementation of MAS and aggressive preoperative and perioperative management led to lower major complication rates and decreased the incidence of readmissions and reoperations after bariatric surgery. PMID- 20005784 TI - Perceived barriers to bariatric surgery among morbidly obese patients. AB - BACKGROUND: Obesity has become a worldwide problem. Surgery has been shown to be a safe and effective therapy. We sought to identify those factors that patients regard as barriers to undergoing a bariatric surgical procedure. METHODS: Morbidly obese patients were asked to complete a 2-page questionnaire during routine outpatient appointments or hospitalization for other reasons. Patients were enrolled from February 2007 to April 2008. The differences between groups were assessed using univariate analysis. RESULTS: A total of 77 patients (41 women and 36 men) were enrolled. Their median age was 51 years, and 49% of the patients were white, followed by Hispanic (23%), and other ethnicities. Of the 77 patients, 9% were supermorbidly obese (body mass index >50 kg/m(2)), and 62% reported having used dieting to lose weight, with greater reports among the women (P = .01). White patients and those >55 years old were more likely to be using some type of weight loss program. Only 40% were physically active. African Americans reported greater rates of regular exercise (P <.01). Of the 77 patients surveyed, 8% had never heard of bariatric surgery. Finally, only 30% of our patient population considered themselves to be morbidly obese. CONCLUSION: The results from the present survey have demonstrated that a lack of insurance coverage is not the main reason for patients not consulting a center to be evaluated for bariatric surgery. Perceived barriers and lack of knowledge exist in both the minds of the general public and physicians. PMID- 20005786 TI - Cystic fibrosis transmembrane conductance regulator (CFTR) regulates the production of osteoprotegerin (OPG) and prostaglandin (PG) E2 in human bone. AB - Bone loss is an important clinical issue in patients with cystic fibrosis (CF). Whether the cystic fibrosis transmembrane conductance regulator (CFTR) plays a direct role in bone cell function is yet unknown. In this study, we provide evidence that inhibition of CFTR-Cl(-) channel function results in a significant decrease of osteoprotegerin (OPG) secretion accompanied with a concomitant increase of prostaglandin (PG) E(2) secretion of primary human osteoblast cultures (n=5). Our data therefore suggest that in bone cells of CF patients, the loss of CFTR activity may result in an increased inflammation-driven bone resorption (through both the reduced OPG and increased PGE(2) production), and thus might contribute to the early bone loss reported in young children with CF. PMID- 20005785 TI - Reduced heart rate variability correlates with insulin resistance but not with measures of obesity in population undergoing laparoscopic Roux-en-Y gastric bypass. AB - BACKGROUND: Obesity is associated with a pathologic predominance of sympathetic over parasympathetic tone. With respect to the heart, this autonomic dysfunction presents as a decreased heart rate variability (HRV), which has been associated with increased cardiovascular morbidity. Gastric bypass (GB) reduces cardiovascular mortality, and, thus, could beneficially affect the HRV. We sought to identify the factors predictive of HRV in a severely obese population of undergoing GB at a university hospital in the United States. METHODS: The data of all patients presenting for GB were included in a prospective database. The homeostatic model of assessment (HOMA) was used to calculate the insulin resistance and glucose disposition index. A 24-hour Holter monitor was used to assess the HRV. Measurements were repeated at 2 weeks and 6 months postoperatively. The correlations between variables were determined using linear mixed models. RESULTS: We studied 30 patients undergoing GB. All exhibited some degree of reduced HRV that improved postoperatively. The HOMA-insulin resistance inversely correlated with the HRV, and the HOMA-glucose disposition index directly correlated with the parameters of HRV in our longitudinal models. Weight, body mass index, excess body weight, gender, and age did not correlate with HRV. Improvements in HRV correlated with reductions in the average heart rate, underscoring a postoperative increase in relative vagal tone. CONCLUSION: HRV in the severely obese is better predicted by the degree of insulin resistance, than by the degree of obesity, age, or gender. GB led to an improvement in HRV, the magnitude of which correlated with the change in insulin resistance and glucose disposition index, but not with weight loss. PMID- 20005788 TI - Multidimensional profiling of components in complex mixtures of natural products for metabolic analysis, proof of concept: application to Quillaja saponins. AB - A method for separation and detection of major and minor components in complex mixtures has been developed, utilising two-dimensional high-performance liquid chromatography (2D-HPLC) combined with electrospray ionisation ion-trap multiple stage mass spectrometry (ESI-ITMS(n)). Chromatographic conditions were matched with mass spectrometric detection to maximise the number of components that could be separated. The described procedure has proven useful to discern several hundreds of saponin components when applied to Quillaja saponaria Molina bark extracts. The discrimination of each saponin component relies on the fact that three coordinates (x, y, z) for each component can be derived from the retention time of the two chromatographic steps (x, y) and the m/z-values from the multiple stage mass spectrometry (z(n), n=1, 2, ...). Thus an improved graphical representation was obtained by combining retention times from the two-stage separation with +MS(1) (z(1)) and the additional structural information from the second mass stage +MS(2) (z(2), z(3)) corresponding to the main fragment ions. By this approach three-dimensional plots can be made that reveal both the chromatographic and structural properties of a specific mixture which can be useful in fingerprinting of complex mixtures. PMID- 20005787 TI - Sensitive determination of carnosine in urine by high-performance liquid chromatography using 4-(5,6-dimethoxy-2-phthalimidinyl)-2-methoxyphenylsulfonyl chloride as a fluorescent labeling reagent. AB - A simple and highly sensitive high-performance liquid chromatography procedure was developed for the determination of carnosine in urine. Carnosine was derivatized with 4-(5,6-dimethoxy-2-phthalimidinyl)-2-methoxyphenylsulfonyl chloride at 70 degrees C for 15 min in borate buffer (20 mmol l(-1), pH 9.0) to produce fluorescent sulfonamides. After hydrolysis of the reaction mixture with formic acid at 100 degrees C for 15 min, the fluorescent derivative of carnosine was separated on a reversed-phase column with a linear gradient elution using solvents of (A) acetate buffer (0.1 mmol l(-1), pH 7.0) and (B) acetonitrile at a flow-rate of 1.0 ml/min and was detected at excitation and emission wavelengths of 318 and 400 nm, respectively. The detection limit of carnosine was 4 fmol at a signal-to-noise ratio of 3. The within-day and day-to-day relative standard deviations were 2.7-4.6% and 0.4-5.2%, respectively. The concentration of carnosine in normal human urine was found to be 4.6-125 nmol (mg creatinine)(-1) (mean+/-SD: 21.6+/-26.6 nmol (mg creatinine)(-1), n=20). PMID- 20005789 TI - Extended applications of distally based axial adipofascial flaps for hand and digits defects. AB - Reverse axial adipofascial flaps use subcutaneous tissue of the laterodigital and dorsal metacarpal or digital areas. They are sited on the arterial branches anastomosing the volar and dorsal arterial networks of the fingers. These flaps allow coverage of wide and distal defects. Metacarpal flaps were used for defects on the proximal phalanx, and digital flaps for the defects over the proximal interphalangeal joint (PIPj) and further distally of the thumb and fingers. A series of 18 cases is reviewed. Dissection of the flap is easy, fast and preserves the collateral nerve and artery of the finger. Skin defects were combined with bone, joint or tendon exposure. The flaps we used were reliable, and bone, joint and tendon reconstruction could be performed at the same time. Procedures were performed in an outpatient setting. Patients were allowed to mobilise the hand as early as possible. The results showed partial distal necrosis in one case and partial loss of the overlying skin graft in two cases, who resolved spontaneously. Donor-site morbidity was minimal. PMID- 20005790 TI - Ensuring safer mandibular angle resection surgery by using a calibrated mandibular angle hook. PMID- 20005791 TI - Stripping of a fistula for complete second branchial cleft. AB - Complete fistulas of the second branchial cleft have a complicated course between the internal and external carotid arteries, crossing the hypoglossal and glossopharyngeal nerve. Therefore, surgical excision implies a tedious dissection of the fistula between the neck vessels and nerves and the multiple or large incisions have major cosmetic consequences. In cases of complete fistulas, we report a more simple method by stripping. Using a guide wire fixed to the first 2cm of the fistula, the fistulous tract is stripped completely inside out to its pharyngeal opening by pulling the oral end of the guide wire. This technique uses a limited skin incision with an excellent cosmetic result and very low morbidity; and furthermore, it shortens the procedure and hospitalisation time. PMID- 20005792 TI - An urgent call to implement systematic monitoring of a comprehensive set of quality indicators for maternity services. AB - This commentary paper proposes that historically the lack of significant system wide reform of the sector may be in part related to the quality of evidence that has informed reviews of maternity services. It calls for the implementation of systematic monitoring of a comprehensive set of quality indicators and makes suggestions about how this goal might be achieved. PMID- 20005793 TI - Switching on epigenetic therapy. AB - Reversible epigenetic changes that alter gene expression are a characteristic of many cancers and other diseases. Biotech companies are taking note and are starting to develop new drugs to reverse such pathogenic "epimutations." PMID- 20005794 TI - iPS cells: mapping the policy issues. AB - Given the explosion of research on induced pluripotent stem (iPS) cells, it is timely to consider the various ethical, legal, and social issues engaged by this fast-moving field. Here, we review issues associated with the procurement, basic research, and clinical translation of iPS cells. PMID- 20005795 TI - The ends have arrived. AB - The 2009 Nobel Prize in Physiology or Medicine has been awarded to Elizabeth Blackburn, Carol Greider, and Jack Szostak for their contributions to our understanding of how the ends of eukaryotic chromosomes, telomeres, are replicated by a specialized reverse transcriptase, telomerase. I present a personal view of the telomere field, putting the contributions of these three Nobel laureates into historical context. PMID- 20005796 TI - The ribosome at atomic resolution. AB - The 2009 Nobel Prize in Chemistry has been awarded to Venki Ramakrishnan, Tom Steitz, and Ada Yonath for crystallographic studies of the ribosome. The atomic resolution structures of the ribosomal subunits provide an extraordinary context for understanding one of the most fundamental aspects of cellular function: protein synthesis. PMID- 20005797 TI - The dangers of transcription. AB - Transcription is obviously essential, but even a good thing can be dangerous at times. In this issue, Lin et al. (2009) provide evidence that binding of the transcription machinery may predispose genome regions to breakage and translocations that may lead to cancer. PMID- 20005798 TI - During transitions proteins make fleeting bonds. AB - How do proteins efficiently and precisely shift from one conformation to another? Gardino et al. (2009) show that transient hydrogen bonds are critical to the conformational transition of the nitrogen regulatory protein NtrC between its native state and its active state. PMID- 20005799 TI - Females battle to suppress their inner male. AB - Conventional wisdom holds that the ovary and testis are terminally differentiated organs in adult mammals. However, Uhlenhaut et al. (2009) now report that deletion of a single gene, Foxl2, is sufficient to induce transdifferentiation of ovary into testis in adult mice, suggesting that testicular development is actively repressed throughout the life of females. PMID- 20005800 TI - A tethering complex recruits SNAREs and grabs vesicles. AB - Protein tethers can bridge gaps between membranes. Ren et al. (2009) now provide evidence that the yeast Dsl1 complex tethers vesicles to the endoplasmic reticulum (ER) by binding ER SNARE proteins at its base and capturing vesicles using a loop region that extends 20 nm from the ER membrane. PMID- 20005801 TI - Wnt signaling and the polarity of the primary body axis. AB - How animals establish and pattern the primary body axis is one of the most fundamental problems in biology. Data from diverse deuterostomes (frog, fish, mouse, and amphioxus) and from planarians (protostomes) suggest that Wnt signaling through beta-catenin controls posterior identity during body plan formation in most bilaterally symmetric animals. Wnt signaling also influences primary axis polarity of pre-bilaterian animals, indicating that an axial patterning role for Wnt signaling predates the evolution of bilaterally symmetric animals. The use of posterior Wnt signaling and anterior Wnt inhibition might be a unifying principle of body plan development in most animals. PMID- 20005802 TI - The structural basis for mRNA recognition and cleavage by the ribosome-dependent endonuclease RelE. AB - Translational control is widely used to adjust gene expression levels. During the stringent response in bacteria, mRNA is degraded on the ribosome by the ribosome dependent endonuclease, RelE. The molecular basis for recognition of the ribosome and mRNA by RelE and the mechanism of cleavage are unknown. Here, we present crystal structures of E. coli RelE in isolation (2.5 A) and bound to programmed Thermus thermophilus 70S ribosomes before (3.3 A) and after (3.6 A) cleavage. RelE occupies the A site and causes cleavage of mRNA after the second nucleotide of the codon by reorienting and activating the mRNA for 2'-OH-induced hydrolysis. Stacking of A site codon bases with conserved residues in RelE and 16S rRNA explains the requirement for the ribosome in catalysis and the subtle sequence specificity of the reaction. These structures provide detailed insight into the translational regulation on the bacterial ribosome by mRNA cleavage. PMID- 20005803 TI - Conserved MicroRNA miR-8/miR-200 and its target USH/FOG2 control growth by regulating PI3K. AB - How body size is determined is a long-standing question in biology, yet its regulatory mechanisms remain largely unknown. Here, we find that a conserved microRNA miR-8 and its target, USH, regulate body size in Drosophila. miR-8 null flies are smaller in size and defective in insulin signaling in fat body that is the fly counterpart of liver and adipose tissue. Fat body-specific expression and clonal analyses reveal that miR-8 activates PI3K, thereby promoting fat cell growth cell-autonomously and enhancing organismal growth non-cell-autonomously. Comparative analyses identify USH and its human homolog, FOG2, as the targets of fly miR-8 and human miR-200, respectively. USH/FOG2 inhibits PI3K activity, suppressing cell growth in both flies and humans. FOG2 directly binds to p85alpha, the regulatory subunit of PI3K, and interferes with the formation of a PI3K complex. Our study identifies two novel regulators of insulin signaling, miR 8/miR-200 and USH/FOG2, and suggests their roles in adolescent growth, aging, and cancer. PMID- 20005804 TI - Transient non-native hydrogen bonds promote activation of a signaling protein. AB - Phosphorylation is a common mechanism for activating proteins within signaling pathways. Yet, the molecular transitions between the inactive and active conformational states are poorly understood. Here we quantitatively characterize the free-energy landscape of activation of a signaling protein, nitrogen regulatory protein C (NtrC), by connecting functional protein dynamics of phosphorylation-dependent activation to protein folding and show that only a rarely populated, pre-existing active conformation is energetically stabilized by phosphorylation. Using nuclear magnetic resonance (NMR) dynamics, we test an atomic scale pathway for the complex conformational transition, inferred from molecular dynamics simulations (Lei et al., 2009). The data show that the loss of native stabilizing contacts during activation is compensated by non-native transient atomic interactions during the transition. The results unravel atomistic details of native-state protein energy landscapes by expanding the knowledge about ground states to transition landscapes. PMID- 20005805 TI - A structure-based mechanism for vesicle capture by the multisubunit tethering complex Dsl1. AB - Vesicle trafficking requires membrane fusion, mediated by SNARE proteins, and upstream events that probably include "tethering," an initial long-range attachment between a vesicle and its target organelle. Among the factors proposed to mediate tethering are a set of multisubunit tethering complexes (MTCs). The Dsl1 complex, with only three subunits, is the simplest known MTC and is essential for the retrograde traffic of COPI-coated vesicles from the Golgi to the ER. To elucidate structural principles underlying MTC function, we have determined the structure of the Dsl1 complex, revealing a tower containing at its base the binding sites for two ER SNAREs and at its tip a flexible lasso for capturing vesicles. The Dsl1 complex binds to individual SNAREs via their N terminal regulatory domains and also to assembled SNARE complexes; moreover, it is capable of accelerating SNARE complex assembly. Our results suggest that even the simplest MTC may be capable of orchestrating vesicle capture, uncoating, and fusion. PMID- 20005806 TI - Somatic sex reprogramming of adult ovaries to testes by FOXL2 ablation. AB - In mammals, the transcription factor SRY, encoded by the Y chromosome, is normally responsible for triggering the indifferent gonads to develop as testes rather than ovaries. However, testis differentiation can occur in its absence. Here we demonstrate in the mouse that a single factor, the forkhead transcriptional regulator FOXL2, is required to prevent transdifferentiation of an adult ovary to a testis. Inducible deletion of Foxl2 in adult ovarian follicles leads to immediate upregulation of testis-specific genes including the critical SRY target gene Sox9. Concordantly, reprogramming of granulosa and theca cell lineages into Sertoli-like and Leydig-like cell lineages occurs with testosterone levels comparable to those of normal XY male littermates. Our results show that maintenance of the ovarian phenotype is an active process throughout life. They might also have important medical implications for the understanding and treatment of some disorders of sexual development in children and premature menopause in women. PMID- 20005807 TI - Platelet polyphosphates are proinflammatory and procoagulant mediators in vivo. AB - Platelets play a central role in thrombosis, hemostasis, and inflammation. We show that activated platelets release inorganic polyphosphate (polyP), a polymer of 60-100 phosphate residues that directly bound to and activated the plasma protease factor XII. PolyP-driven factor XII activation triggered release of the inflammatory mediator bradykinin by plasma kallikrein-mediated kininogen processing. PolyP increased vascular permeability and induced fluid extravasation in skin microvessels of mice. Mice deficient in factor XII or bradykinin receptors were resistant to polyP-induced leakage. PolyP initiated clotting of plasma via the contact pathway. Ablation of intrinsic coagulation pathway proteases factor XII and factor XI protected mice from polyP-triggered lethal pulmonary embolism. Targeting polyP with phosphatases interfered with procoagulant activity of activated platelets and blocked platelet-induced thrombosis in mice. Addition of polyP restored defective plasma clotting of Hermansky-Pudlak Syndrome patients, who lack platelet polyP. The data identify polyP as a new class of mediator having fundamental roles in platelet-driven proinflammatory and procoagulant disorders. PMID- 20005808 TI - Reduced IGF-1 signaling delays age-associated proteotoxicity in mice. AB - The insulin/insulin growth factor (IGF) signaling (IIS) pathway is a key regulator of aging of worms, flies, mice, and likely humans. Delayed aging by IIS reduction protects the nematode C. elegans from toxicity associated with the aggregation of the Alzheimer's disease-linked human peptide, Abeta. We reduced IGF signaling in Alzheimer's model mice and discovered that these animals are protected from Alzheimer's-like disease symptoms, including reduced behavioral impairment, neuroinflammation, and neuronal loss. This protection is correlated with the hyperaggregation of Abeta leading to tightly packed, ordered plaques, suggesting that one aspect of the protection conferred by reduced IGF signaling is the sequestration of soluble Abeta oligomers into dense aggregates of lower toxicity. These findings indicate that the IGF signaling-regulated mechanism that protects from Abeta toxicity is conserved from worms to mammals and point to the modulation of this signaling pathway as a promising strategy for the development of Alzheimer's disease therapy. PMID- 20005809 TI - Prediction of photoperiodic regulators from quantitative gene circuit models. AB - Photoperiod sensors allow physiological adaptation to the changing seasons. The prevalent hypothesis is that day length perception is mediated through coupling of an endogenous rhythm with an external light signal. Sufficient molecular data are available to test this quantitatively in plants, though not yet in mammals. In Arabidopsis, the clock-regulated genes CONSTANS (CO) and FLAVIN, KELCH, F-BOX (FKF1) and their light-sensitive proteins are thought to form an external coincidence sensor. Here, we model the integration of light and timing information by CO, its target gene FLOWERING LOCUS T (FT), and the circadian clock. Among other predictions, our models show that FKF1 activates FT. We demonstrate experimentally that this effect is independent of the known activation of CO by FKF1, thus we locate a major, novel controller of photoperiodism. External coincidence is part of a complex photoperiod sensor: modeling makes this complexity explicit and may thus contribute to crop improvement. PMID- 20005810 TI - Little effect of the tan locus on pigmentation in female hybrids between Drosophila santomea and D. melanogaster. AB - Previous work on Drosophila santomea suggested that its absence of abdominal pigmentation, compared to the other darkly pigmented species, is based on mutations in the cis-regulatory region of tan, inactivating the expression of that gene in the abdomen of D. santomea males and females. Our discovery that D. santomea males can produce viable hybrids when mated to D. melanogaster females enables us to use the armamentarium of genetic tools in the latter species to study the genetic basis of this interspecific difference in pigmentation. Hybridization tests using D. melanogaster deficiencies that include tan show no evidence that this locus is involved in the lighter pigmentation of D. santomea females; rather, the pigmentation difference appears to involve at least four other loci in the region. Earlier results implicating tan may have been based on a type of transgenic analysis that can give misleading results about the genes involved in an evolutionary change. PMID- 20005812 TI - Enhanced SnapShot: endosome-to-golgi retrieval. PMID- 20005811 TI - Evolution of the tan locus contributed to pigment loss in Drosophila santomea: a response to Matute et al. AB - We have shown previously that the loss of abdominal pigmentation in D. santomea relative to its sister species D. yakuba resulted, in part, from cis-regulatory mutations at the tan locus. Matute et al. claim, based solely upon extrapolation from genetic crosses of D. santomea and D. melanogaster, a much more divergent species, that at least four X chromosome regions but not tan are responsible for pigmentation differences. Here, we provide additional evidence from introgressions of D. yakuba genes into D. santomea that support a causative role for tan in the loss of pigmentation and present analyses that contradict Matute et al.'s claims. We discuss how the choice of parental species and other factors affect the ability to identify loci responsible for species divergence, and we affirm that all of our previously reported results and conclusions stand. PMID- 20005813 TI - Taking off the SOCS: cytokine signaling spurs regeneration. AB - Strategies to improve function after CNS injuries must contend with the failure of axons to regrow after transection in adult mammals. In this issue of Neuron, Smith et al. provide an important advance by demonstrating that SOCS3 acts as a key negative regulator of adult optic nerve regeneration. PMID- 20005814 TI - A new class of spinal interneurons: the origin and function of C boutons is solved. AB - C boutons, a major synaptic input to motor neurons, were first described years ago, but their origin and functional significance were unknown. In this issue of Neuron, a group of cholinergic interneurons located near the central canal is identified as the source of C boutons. The rhythmic activity in these neurons during locomotion increases motor neuronal excitability, thereby potentiating the strength of muscle contraction. PMID- 20005815 TI - Rab3 GTPase lands Bruchpilot. AB - Active zones are the sites of neurotransmitter release, but their assembly mechanisms are poorly understood. In this issue of Neuron, Graf et al. perform a genetic screen in Drosophila and uncover a novel role for the Rab3 GTPase in organizing the active zone at the neuromuscular junction. PMID- 20005816 TI - Fields of gain in the brain. AB - For more than two decades, neuroscientists have debated the role of "gain fields" in sensorimotor transformations. In this issue of Neuron, Chang et al. demonstrate a tight correlation between eye and hand position gain fields in the "parietal reach region," strongly suggesting that they play a functional role in computing the reach command. PMID- 20005817 TI - The model- and the data-gamma. AB - Brain regions that are structurally connected might become functionally connected by synchronization in the gamma-frequency band. In a recent issue of Nature, Colgin et al. show spatially and temporally fine-grained gamma-band synchronization between different parts of the rat hippocampal formation, suggesting a theta-modulated switching of gamma-mediated communication. PMID- 20005818 TI - The challenges natural images pose for visual adaptation. AB - Advances in our understanding of natural image statistics and of gain control within the retinal circuitry are leading to new insights into the classic problem of retinal light adaptation. Here we review what we know about how rapid adaptation occurs during active exploration of the visual scene. Adaptational mechanisms must balance the competing demands of adapting quickly, locally, and reliably, and this balance must be maintained as lighting conditions change. Multiple adaptational mechanisms in different locations within the retina act in concert to accomplish this task, with lighting conditions dictating which mechanisms dominate. PMID- 20005819 TI - SOCS3 deletion promotes optic nerve regeneration in vivo. AB - Axon regeneration failure accounts for permanent functional deficits following CNS injury in adult mammals. However, the underlying mechanisms remain elusive. In analyzing axon regeneration in different mutant mouse lines, we discovered that deletion of suppressor of cytokine signaling 3 (SOCS3) in adult retinal ganglion cells (RGCs) promotes robust regeneration of injured optic nerve axons. This regeneration-promoting effect is efficiently blocked in SOCS3-gp130 double knockout mice, suggesting that SOCS3 deletion promotes axon regeneration via a gp130-dependent pathway. Consistently, a transient upregulation of ciliary neurotrophic factor (CNTF) was observed within the retina following optic nerve injury. Intravitreal application of CNTF further enhances axon regeneration from SOCS3-deleted RGCs. Together, our results suggest that compromised responsiveness to injury-induced growth factors in mature neurons contributes significantly to regeneration failure. Thus, developing strategies to modulate negative signaling regulators may be an efficient strategy of promoting axon regeneration after CNS injury. PMID- 20005820 TI - Altering cortical connectivity: remediation-induced changes in the white matter of poor readers. AB - Neuroimaging studies using diffusion tensor imaging (DTI) have revealed regions of cerebral white matter with decreased microstructural organization (lowerfractional anisotropy or FA) among poor readers. We examined whether 100 hr of intensive remedial instruction affected the white matter of 8- to 10-year-old poor readers. Prior to instruction, poor readers had significantly lower FA than good readers in a region of the left anterior centrum semiovale. The instruction resulted in a change in white matter (significantly increased FA), and in the very same region. The FA increase was correlated with a decrease in radial diffusivity (but not with a change in axial diffusivity), suggesting that myelination had increased. Furthermore, the FA increase was correlated with improvement in phonological decoding ability, clarifying the cognitive locus of the effect. The results demonstrate the capability of a behavioral intervention to bring about a positive change in cortico-cortical white matter tracts. PMID- 20005821 TI - Overexpression of low-density lipoprotein receptor in the brain markedly inhibits amyloid deposition and increases extracellular A beta clearance. AB - Apolipoprotein E (APOE) is the strongest genetic risk factor for Alzheimer's disease (AD). Previous studies suggest that the effect of apoE on amyloid-beta (A beta) accumulation plays a major role in AD pathogenesis. Therefore, understanding proteins that control apoE metabolism may provide new targets for regulating A beta levels. LDLR, a member of the LDL receptor family, binds to apoE, yet its potential role in AD pathogenesis remains unclear. We hypothesized that LDLR overexpression in the brain would decrease apoE levels, enhance A beta clearance, and decrease A beta deposition. To test our hypothesis, we created several transgenic mice that overexpress LDLR in the brain and found that apoE levels in these mice decreased by 50%-90%. Furthermore, LDLR overexpression dramatically reduced A beta aggregation and enhanced A beta clearance from the brain extracellular space. Plaque-associated neuroinflammatory responses were attenuated in LDLR transgenic mice. These findings suggest that increasing LDLR levels may represent a novel AD treatment strategy. PMID- 20005822 TI - A cluster of cholinergic premotor interneurons modulates mouse locomotor activity. AB - Mammalian motor programs are controlled by networks of spinal interneurons that set the rhythm and intensity of motor neuron firing. Motor neurons have long been known to receive prominent "C bouton" cholinergic inputs from spinal interneurons, but the source and function of these synaptic inputs have remained obscure. We show here that the transcription factor Pitx2 marks a small cluster of spinal cholinergic interneurons, V0(C) neurons, that represents the sole source of C bouton inputs to motor neurons. The activity of these cholinergic interneurons is tightly phase locked with motor neuron bursting during fictive locomotor activity, suggesting a role in the modulation of motor neuron firing frequency. Genetic inactivation of the output of these neurons impairs a locomotor task-dependent increase in motor neuron firing and muscle activation. Thus, V0(C) interneurons represent a defined class of spinal cholinergic interneurons with an intrinsic neuromodulatory role in the control of locomotor behavior. PMID- 20005823 TI - Rab3 dynamically controls protein composition at active zones. AB - Synaptic transmission requires the localization of presynaptic release machinery to active zones. Mechanisms regulating the abundance of such synaptic proteins at individual release sites are likely determinants of site-specific synaptic efficacy. We now identify a role for the small GTPase Rab3 in regulating the distribution of presynaptic components to active zones. At Drosophila rab3 mutant NMJs, the presynaptic protein Bruchpilot, calcium channels, and electron-dense T bars are concentrated at a fraction of available active zones, leaving the majority of sites devoid of these key presynaptic release components. Late addition of Rab3 to mutant NMJs rapidly reverses this phenotype by recruiting Brp to sites previously lacking the protein, demonstrating that Rab3 can dynamically control the composition of the presynaptic release machinery. While previous studies of Rab3 have focused on its role in the synaptic vesicle cycle, these findings demonstrate an additional and unexpected function for Rab3 in the localization of presynaptic proteins to active zones. PMID- 20005825 TI - Frequency transitions in odor-evoked neural oscillations. AB - In many species, sensory stimuli elicit the oscillatory synchronization of groups of neurons. What determines the properties of these oscillations? In the olfactory system of the moth, we found that odors elicited oscillatory synchronization through a neural mechanism like that described in locust and Drosophila. During responses to long odor pulses, oscillations suddenly slowed as net olfactory receptor neuron (ORN) output decreased; thus, stimulus intensity appeared to determine oscillation frequency. However, changing the concentration of the odor had little effect upon oscillatory frequency. Our recordings in vivo and computational models based on these results suggested that the main effect of increasing odor concentration was to recruit additional, less well-tuned ORNs whose firing rates were tightly constrained by adaptation and saturation. Thus, in the periphery, concentration is encoded mainly by the size of the responsive ORN population, and oscillation frequency is set by the adaptation and saturation of this response. PMID- 20005824 TI - Control of cognition and adaptive behavior by the GLP/G9a epigenetic suppressor complex. AB - The genetic basis of cognition and behavioral adaptation to the environment remains poorly understood. Here we demonstrate that the histone methyltransferase complex GLP/G9a controls cognition and adaptive responses in a region-specific fashion in the adult brain. Using conditional mutagenesis in mice, we show that postnatal, neuron-specific deficiency of GLP/G9a leads to derepression of numerous nonneuronal and neuron progenitor genes in adult neurons. This transcriptional alteration is associated with complex behavioral abnormalities, including defects in learning, motivation, and environmental adaptation. The behavioral changes triggered by GLP/G9a deficiency are similar to key symptoms of the human 9q34 mental retardation syndrome that is associated with structural alterations of the GLP/EHMT1 gene. The likely causal role of GLP/G9a in mental retardation in mice and humans suggests a key role for the GLP/G9a-controlled histone H3K9 dimethylation in regulation of brain function through maintenance of the transcriptional homeostasis in adult neurons. PMID- 20005826 TI - Functional masking of deprived eye responses by callosal input during ocular dominance plasticity. AB - Monocular deprivation (MD) is a well-known paradigm of experience-dependent plasticity in which cortical neurons exhibit a shift of ocular dominance (OD) toward the open eye. The mechanisms underlying this form of plasticity are incompletely understood. Here we demonstrate the involvement of callosal connections in the synaptic modifications occurring during MD. Rats at the peak of the critical period were deprived for 7 days, resulting in the expected OD shift toward the open eye. Acute microinjection of the activity blocker muscimol into the visual cortex contralateral to the recording site restored binocularity of cortical cells. Continuous silencing of callosal input throughout the period of MD also resulted in substantial attenuation of the OD shift. Blockade of interhemispheric communication selectively enhanced deprived eye responses with no effect on open eye-driven activity. We conclude that callosal inputs play a key role in functional weakening of less active connections during OD plasticity. PMID- 20005827 TI - Rapid axonal sprouting and pruning accompany functional reorganization in primary visual cortex. AB - The functional architecture of adult cerebral cortex retains a capacity for experience-dependent change. This is seen following focal binocular lesions, which induce rapid changes in receptive field size and position. To follow the dynamics of the circuitry underlying these changes, we imaged the intrinsic long range horizontal connections within the lesion projection zone (LPZ) in adult macaque primary visual cortex. To image the same axons over time, we combined viral vector-mediated EGFP transfer and two-photon microscopy. The lesion triggered, within the first week, an approximately 2-fold outgrowth of axons toward the center of the LPZ. Over the subsequent month, axonal density declined due to a parallel process of pruning and sprouting but maintained a net increase relative to prelesion levels. The rate of turnover of axonal boutons also increased. The axonal restructuring recapitulates the pattern of exuberance and pruning seen in early development and correlates well with the functional changes following retinal lesions. PMID- 20005828 TI - Flexibility of sensory representations in prefrontal cortex depends on cell type. AB - Discrimination tasks require processing, interpreting, and linking sensory information to the appropriate motor response. We report that neurons in prefrontal cortex (PFC) represent visual motion with precision comparable to cortical neurons at early stages of motion processing, and readily adapt this representation to behavioral context. We found that direction selectivity, recorded while the monkeys discriminated directions, decreased when they judged motion speed and ignored its direction. This decrease was more pronounced in neurons classified as narrow-spiking (NS) putative interneurons than in broad spiking (BS) putative pyramidal neurons. However, during passive fixation, when the link between motion and its behavioral relevance was removed, both cell types showed a severe selectivity loss. Our results show that flexible sensory representation during active discrimination tasks is achieved in the PFC by a specialized neuronal network of both NS neurons readily adjusting their selectivity to behavioral context, and BS neurons capable of maintaining relatively stable sensory representation. PMID- 20005830 TI - The neural circuitry of a broken promise. AB - Promises are one of the oldest human-specific psychological mechanisms fostering cooperation and trust. Here, we study the neural underpinnings of promise keeping and promise breaking. Subjects first make a promise decision (promise stage), then they anticipate whether the promise affects the interaction partner's decision (anticipation stage) and are subsequently free to keep or break the promise (decision stage). Findings revealed that the breaking of the promise is associated with increased activation in the DLPFC, ACC, and amygdala, suggesting that the dishonest act involves an emotional conflict due to the suppression of the honest response. Moreover, the breach of the promise can be predicted by a perfidious brain activity pattern (anterior insula, ACC, inferior frontal gyrus) during the promise and anticipation stage, indicating that brain measurements may reveal malevolent intentions before dishonest or deceitful acts are actually committed. PMID- 20005831 TI - Reverse transcribing the code for chromosome stability. AB - The linearity of eukaryotic chromosomes presents challenges to cells, as the presence of DNA "ends" poses problems for the DNA replication machinery and the cell's damage response systems. This year's Nobel Prize in Physiology or Medicine recognized groundbreaking studies establishing the telomere field as a crucial area of biomedical research. PMID- 20005829 TI - Using a compound gain field to compute a reach plan. AB - A gain field, the scaling of a tuned neuronal response by a postural signal, may help support neuronal computation. Here, we characterize eye and hand position gain fields in the parietal reach region (PRR). Eye and hand gain fields in individual PRR neurons are similar in magnitude but opposite in sign to one another. This systematic arrangement produces a compound gain field that is proportional to the distance between gaze location and initial hand position. As a result, the visual response to a target for an upcoming reach is scaled by the initial gaze-to-hand distance. Such a scaling is similar to what would be predicted in a neural network that mediates between eye- and hand-centered representations of target location. This systematic arrangement supports a role of PRR in visually guided reaching and provides strong evidence that gain fields are used for neural computations. PMID- 20005832 TI - Resolving the elegant architecture of the ribosome. AB - This year's Nobel Prize in Chemistry rewards Ada Yonath, Tom Steitz, and Venki Ramakrishnan for their groundbreaking structural studies on the ribosome. PMID- 20005833 TI - Signaling motifs and Weber's law. AB - New experimental and theoretical studies reported by Uri Alon, Marc Kirschner, and colleagues in this issue of Molecular Cell suggest that Weber's law of sensory perception may apply to a number of cell signaling processes. PMID- 20005834 TI - Hydroxyurea triggers cellular responses that actively cause bacterial cell death. AB - In this issue of Molecular Cell, Davies et al. (2009) work out a sequence of active cellular events that lead to the death of Escherichia coli in the presence of the drug hydroxyurea. PMID- 20005835 TI - Evolving protein stability through genetic selection. AB - In this issue of Molecular Cell, Foit et al. (2009) probe cellular protein folding using a split beta-lactamase approach for evolving protein stability in the absence of any requirement for function. PMID- 20005836 TI - A FIRE-y PAGE in the computational analysis of cancer profiles. AB - In this issue of Molecular Cell, Goodarzi et al. (2009) employ novel computational approaches to demonstrate the power of global systems biology analyses in elucidating cancer biology. PMID- 20005837 TI - Checkpoint Mec-tivation comes in many flavors. AB - In this issue of Molecular Cell, Navadgi-Patil and Burgers show that the budding yeast Mec1 kinase is activated by DNA damage through two distinct Ddc1-mediated mechanisms during G1 and G2 phases of the cell cycle. PMID- 20005838 TI - A tangled web of ubiquitin chains: breaking news in TNF-R1 signaling. AB - A flurry of recent revelations is challenging the current dogma on how ubiquitin dependent processes culminate in the activation of NF-kappaB by TNF. Here, we integrate these findings into a model for TNF-R1 signaling-and underscore the importance of individual components, including linear ubiquitin chains-which allows for the remarkable versatility of the ubiquitin system. PMID- 20005840 TI - SOCS1 links cytokine signaling to p53 and senescence. AB - SOCS1 is lost in many human tumors, but its tumor suppression activities are not well understood. We report that SOCS1 is required for transcriptional activity, DNA binding, and serine 15 phosphorylation of p53 in the context of STAT5 signaling. In agreement, inactivation of SOCS1 disabled p53-dependent senescence in response to oncogenic STAT5A and radiation-induced apoptosis in T cells. In addition, SOCS1 was sufficient to induce p53-dependent senescence in fibroblasts. The mechanism of activation of p53 by SOCS1 involved a direct interaction between the SH2 domain of SOCS1 and the N-terminal transactivation domain of p53, while the C-terminal domain of SOCS1 containing the SOCS Box mediated interaction with the DNA damage-regulated kinases ATM/ATR. Also, SOCS1 colocalized with ATM at DNA damage foci induced by oncogenic STAT5A. Collectively, these results add another component to the p53 and DNA damage networks and reveal a mechanism by which SOCS1 functions as a tumor suppressor. PMID- 20005839 TI - The unstructured C-terminal tail of the 9-1-1 clamp subunit Ddc1 activates Mec1/ATR via two distinct mechanisms. AB - DNA damage checkpoint pathways operate to prevent cell-cycle progression in response to DNA damage and replication stress. In S. cerevisiae, Mec1-Ddc2 (human ATR-ATRIP) is the principal checkpoint protein kinase. Biochemical studies have identified two factors, the 9-1-1 checkpoint clamp and the Dpb11/TopBP1 replication protein, as potential activators of Mec1/ATR. Here, we show that G1 phase checkpoint activation of Mec1 is achieved by the Ddc1 subunit of 9-1-1, while Dpb11 is dispensable. However, in G2, 9-1-1 activates Mec1 by two distinct mechanisms. One mechanism involves direct activation of Mec1 by Ddc1, while the second proceeds by Dpb11 recruitment mediated through Ddc1 T602 phosphorylation. Two aromatic residues, W352 and W544, localized to two widely separated, conserved motifs of Ddc1, are essential for Mec1 activation in vitro and checkpoint function in G1. Remarkably, small peptides that fuse the two tryptophan-containing motifs together are proficient in activating Mec1. PMID- 20005842 TI - Usa1 functions as a scaffold of the HRD-ubiquitin ligase. AB - Protein quality control in the endoplasmic reticulum is of central importance for cellular homeostasis in eukaryotes. Crucial for this process is the HRD-ubiquitin ligase (HMG-CoA reductase degradation), which singles out terminally misfolded proteins and routes them for degradation to cytoplasmic 26S-proteasomes. Certain functions of this enzyme complex are allocated to defined subunits. However, it remains unclear how these components act in a concerted manner. Here, we show that Usa1 functions as a major scaffold protein of the HRD-ligase. For the turnover of soluble substrates, Der1 binding to the C terminus of Usa1 is required. The N terminus of Usa1 associates with Hrd1 and thus bridges Der1 to Hrd1. Strikingly, the Usa1 N terminus also induces oligomerization of the HRD complex, which is an exclusive prerequisite for the degradation of membrane proteins. Our data demonstrate that scaffold proteins are required to adapt ubiquitin ligase activities toward different classes of substrates. PMID- 20005841 TI - Rrp17p is a eukaryotic exonuclease required for 5' end processing of Pre-60S ribosomal RNA. AB - Ribosomal processing requires a series of endo- and exonucleolytic steps for the production of mature ribosomes, of which most have been described. To ensure ribosome synthesis, 3' end formation of rRNA uses multiple nucleases acting in parallel; however, a similar parallel mechanism had not been described for 5' end maturation. Here, we identify Rrp17p as a previously unidentified 5'-3' exonuclease essential for ribosome biogenesis, functioning with Rat1p in a parallel processing pathway analogous to that of 3' end formation. Rrp17p is required for efficient exonuclease digestion of the mature 5' ends of 5.8S(S) and 25S rRNAs, contains a catalytic domain close to its N terminus, and is highly conserved among higher eukaryotes, being a member of a family of exonucleases. We show that Rrp17p binds late pre-60S ribosomes, accompanying them from the nucleolus to the nuclear periphery, and provide evidence for physical and functional links between late 60S subunit processing and export. PMID- 20005843 TI - Ubiquitinated proteins activate the proteasome by binding to Usp14/Ubp6, which causes 20S gate opening. AB - In eukaryotic cells, ubiquitination of proteins leads to their degradation by the 26S proteasome. We tested if the ubiquitin (Ub) chain also regulates the proteasome's capacity for proteolysis. After incubation with polyubiquitinated proteins, 26S proteasomes hydrolyzed peptides and proteins 2- to 7-fold faster. Ub conjugates enhanced peptide hydrolysis by stimulating gate opening in the 20S proteasome. This stimulation was seen when this gate was closed or transiently open, but not maximally open. Gate opening requires conjugate association with Usp14/Ubp6 and also occurs if Ub aldehyde occupies this isopeptidase's active site. No stimulation was observed with 26S from Ubp6Delta mutants, but this effect was restored upon addition of Usp14/Ubp6 (even an inactive Ubp6). The stimulation of gate opening by Ub conjugates through Usp14/Ubp6 requires nucleotide binding to the gate-regulatory ATPases. This activation enhances the selectivity of the 26S proteasome for ubiquitinated proteins and links their deubiquitination to their degradation. PMID- 20005844 TI - Ubiquitin-like sequence in ASK1 plays critical roles in the recognition and stabilization by USP9X and oxidative stress-induced cell death. AB - Ubiquitination is an important posttranslational modification that regulates various cellular processes, including signal transduction. However, physiological roles of ubiquitination in the regulation of MAPK pathways are poorly understood. Here, we identified the deubiquitinating enzyme USP9X as a binding partner of ASK1 that mediates oxidative stress-induced cell death through activation of the JNK and p38 MAPK pathways. In the recognition of ubiquitin by deubiquitinating enzymes, the importance of a tandem glycine-glycine sequence in the ubiquitin C terminus has been suggested. Interestingly, ASK1 contains six amino acids identical to the ubiquitin C terminus (LRLRGG), and the GG sequence of ASK1 was required for the USP9X-ASK1 interaction. We also found that USP9X interacted with oxidative stress-activated ASK1 and prevented it from undergoing ubiquitin dependent degradation. In USP9X-deficient cells, oxidative stress-induced JNK activation and subsequent cell death were reduced. These results demonstrate that USP9X-dependent stabilization of activated ASK1 plays a crucial role in oxidative stress-induced cell death. PMID- 20005845 TI - The pseudoactive site of ILK is essential for its binding to alpha-Parvin and localization to focal adhesions. AB - Integrin-linked kinase (ILK) plays a pivotal role in connecting transmembrane receptor integrin to the actin cytoskeleton and thereby regulating diverse cell adhesion-dependent processes. The kinase domain (KD) of ILK is indispensable for its function, but the underlying molecular basis remains enigmatic. Here we present the crystal structure of the ILK KD bound to its cytoskeletal regulator, the C-terminal calponin homology domain of alpha-parvin. While maintaining a canonical kinase fold, the ILK KD displays a striking pseudoactive site conformation. We show that rather than performing the kinase function, this conformation specifically recognizes alpha-parvin for promoting effective assembly of ILK into focal adhesions. The alpha-parvin-bound ILK KD can simultaneously engage integrin beta cytoplasmic tails. These results thus define ILK as a distinct pseudokinase that mechanically couples integrin and alpha parvin for mediating cell adhesion. They also highlight functional diversity of the kinase fold and its "active" site in mediating many biological processes. PMID- 20005846 TI - Recruitment of the linear ubiquitin chain assembly complex stabilizes the TNF-R1 signaling complex and is required for TNF-mediated gene induction. AB - TNF is a key inflammatory cytokine. Using a modified tandem affinity purification approach, we identified HOIL-1 and HOIP as functional components of the native TNF-R1 signaling complex (TNF-RSC). Together, they were shown to form a linear ubiquitin chain assembly complex (LUBAC) and to ubiquitylate NEMO. We show that LUBAC binds to ubiquitin chains of different linkage types and that its recruitment to the TNF-RSC is impaired in TRADD-, TRAF2-, and cIAP1/2- but not in RIP1- or NEMO-deficient MEFs. Furthermore, the E3 ligase activity of cIAPs, but not TRAF2, is required for HOIL-1 recruitment to the TNF-RSC. LUBAC enhances NEMO interaction with the TNF-RSC, stabilizes this protein complex, and is required for efficient TNF-induced activation of NF-kappaB and JNK, resulting in apoptosis inhibition. Finally, we demonstrate that sustained stability of the TNF-RSC requires LUBAC's enzymatic activity, thereby adding a third form of ubiquitin linkage to the triggering of TNF signaling by the TNF-RSC. PMID- 20005847 TI - Hydroxyurea induces hydroxyl radical-mediated cell death in Escherichia coli. AB - Hydroxyurea (HU) specifically inhibits class I ribonucleotide reductase (RNR), depleting dNTP pools and leading to replication fork arrest. Although HU inhibition of RNR is well recognized, the mechanism by which it leads to cell death remains unknown. To investigate the mechanism of HU-induced cell death, we used a systems-level approach to determine the genomic and physiological responses of E. coli to HU treatment. Our results suggest a model by which HU treatment rapidly induces a set of protective responses to manage genomic instability. Continued HU stress activates iron uptake and toxins MazF and RelE, whose activity causes the synthesis of incompletely translated proteins and stimulation of envelope stress responses. These effects alter the properties of one of the cell's terminal cytochrome oxidases, causing an increase in superoxide production. The increased superoxide production, together with the increased iron uptake, fuels the formation of hydroxyl radicals that contribute to HU-induced cell death. PMID- 20005848 TI - Optimizing protein stability in vivo. AB - Identifying mutations that stabilize proteins is challenging because most substitutions are destabilizing. In addition to being of immense practical utility, the ability to evolve protein stability in vivo may indicate how evolution has formed today's protein sequences. Here we describe a genetic selection that directly links the in vivo stability of proteins to antibiotic resistance. It allows the identification of stabilizing mutations within proteins. The large majority of mutants selected for improved antibiotic resistance are stabilized both thermodynamically and kinetically, indicating that similar principles govern stability in vivo and in vitro. The approach requires no prior structural or functional knowledge and allows selection for stability without a need to maintain function. Mutations that enhance thermodynamic stability of the protein Im7 map overwhelmingly to surface residues involved in binding to colicin E7, showing how the evolutionary pressures that drive Im7-E7 complex formation have compromised the stability of the isolated Im7 protein. PMID- 20005849 TI - Evidence that fold-change, and not absolute level, of beta-catenin dictates Wnt signaling. AB - In response to Wnt stimulation, beta-catenin accumulates and activates target genes. Using modeling and experimental analysis, we found that the level of beta catenin is sensitive to perturbations in the pathway, such that cellular variation would be expected to alter the signaling outcome. One unusual parameter was robust: the fold-change in beta-catenin level (post-Wnt/pre-Wnt). In Xenopus, dorsal-anterior development and target gene expression are robust to perturbations that alter the final level but leave the fold-change intact. These suggest, first, that despite cellular noise, the cell responds reliably to Wnt stimulation by maintaining a robust fold-change in beta-catenin. Second, the transcriptional machinery downstream of the Wnt pathway does not simply read the beta-catenin level after Wnt stimulation but computes fold-changes in beta catenin. Analogous to Weber's Law in sensory physiology, some gene transcription networks must respond to fold-changes in signals, rather than absolute levels, which may buffer stochastic, genetic, and environmental variation. PMID- 20005850 TI - Dynamics and variability of ERK2 response to EGF in individual living cells. AB - Signal-transduction cascades are usually studied on cell averages, masking variability between individual cells. To address this, we studied in individual cells the dynamic response of ERK2, a well-characterized MAPK signaling protein, which enters the nucleus upon stimulation. Using fluorescent tagging at the endogenous chromosomal locus, we found that cells show wide basal variation in ERK2 nuclear levels. Upon EGF stimulation, cells show (1) a fold-change response, where peak nuclear accumulation of ERK2 is proportional to basal level in each cell; and (2) exact adaptation in nuclear levels of ERK2, returning to original basal level of each cell. The timing of ERK2 dynamics is more precise between cells than its amplitude. We further found that in some cells ERK2 exhibits a second pulse of nuclear entry, smaller than the first. The present study suggests that this signaling system compensates for natural biological noise: despite large variation in nuclear basal levels, ERK2's fold dynamics is similar between cells. PMID- 20005851 TI - The incoherent feedforward loop can provide fold-change detection in gene regulation. AB - Many sensory systems (e.g., vision and hearing) show a response that is proportional to the fold-change in the stimulus relative to the background, a feature related to Weber's Law. Recent experiments suggest such a fold-change detection feature in signaling systems in cells: a response that depends on the fold-change in the input signal, and not on its absolute level. It is therefore of interest to find molecular mechanisms of gene regulation that can provide such fold-change detection. Here, we demonstrate theoretically that fold-change detection can be generated by one of the most common network motifs in transcription networks, the incoherent feedforward loop (I1-FFL), in which an activator regulates both a gene and a repressor of the gene. The fold-change detection feature of the I1-FFL applies to the entire shape of the response, including its amplitude and duration, and is valid for a wide range of biochemical parameters. PMID- 20005852 TI - Revealing global regulatory perturbations across human cancers. AB - The discovery of pathways and regulatory networks whose perturbation contributes to neoplastic transformation remains a fundamental challenge for cancer biology. We show that such pathway perturbations, and the cis-regulatory elements through which they operate, can be efficiently extracted from global gene expression profiles. Our approach utilizes information-theoretic analysis of expression levels, pathways, and genomic sequences. Analysis across a diverse set of human cancers reveals the majority of previously known cancer pathways. Through de novo motif discovery we associate these pathways with transcription-factor binding sites and miRNA targets, including those of E2F, NF-Y, p53, and let-7. Follow-up experiments confirmed that these predictions correspond to functional in vivo regulatory interactions. Strikingly, the majority of the perturbations, associated with putative cis-regulatory elements, fall outside of known cancer pathways. Our study provides a systems-level dissection of regulatory perturbations in cancer-an essential component of a rational strategy for therapeutic intervention and drug-target discovery. PMID- 20005853 TI - Enhancement of IL-10 bioactivity using an IL-10 peptide-based vaccine exacerbates Leishmania major infection and improves airway inflammation in mice. AB - IL-10 is a regulatory cytokine that plays important roles in promoting disease progression in cutaneous leishmaniasis and suppressing allergic responses in asthma. We sought to develop an IL-10 peptide-based vaccine for the control of IL 10-related diseases. To break self-tolerance, hepatitis B core antigen (HBcAg) was used as a carrier. The vaccine was prepared by inserting a peptide derived from mouse IL-10 into the carrier using gene recombination methods. This vaccine presented as virus-like particles, bound to polyclonal anti-IL-10 antibodies, and induced high titers of IL-10-specific IgG. The in vivo effects of the vaccine were investigated in a murine model of cutaneous leishmaniasis. Unexpectedly, vaccinated mice developed larger cutaneous lesions, harbored significantly more parasites, and cells from lymph nodes produced higher amounts of parasite specific IL-4, IL-10 and IFN-gamma in cultures. Further in vitro studies showed that serum IL-10-specific IgG from vaccinated mice significantly enhanced IL-10 bioactivity dose-dependently. This enhancing effect was confirmed in OVA-induced asthmatic mice. Vaccinated mice exhibited a significant decrease in airway eosinophils, lung inflammation, goblet hyperplasia, and levels of serum OVA specific IgE, compared to control mice. We concluded that the IL-10 vaccine enhances the bioactivity of IL-10 in vitro and in vivo, providing a potential therapeutic approach in diseases associated with insufficient IL-10 production. PMID- 20005854 TI - Evaluation of the immunogenicity of a recombinant glycoprotein-based Chandipura vaccine in combination with commercially available DPT vaccine. AB - Chandipura virus (CHPV) belongs to family Rhabdovoridae and has emerged as an encephalitis causing pathogen with high mortality among pediatric population from three Indian states. The recombinant glycoprotein (rGp) was shown to be an excellent vaccine candidate as evaluated in a murine model. As the disease is predominantly rural, to ensure maximum coverage for Chandipura vaccine, an attempt was made to evaluate combination of rGp and a commercially available DPT vaccine (CHP-DPT). When CHP-DPT was used for immunization of mice, 90% seroconversion against rGp with high antibody titers (1:1200 by ELISA and 1:320 by neutralization test) was observed and did not differ from mice immunized with rGp alone (P>0.05). Similarly seroconversions and antibody titers against DPT were comparable in mice immunized with DPT alone or in combination with rGp. Seroconversions and antibody titers ranged from 90 to 100% and 1:1200 to 1:2400 respectively. Intracerebral challenge with homologous CHPV strain resulted in 90% survival in rGp alone and CHP-DPT groups. Lymphocyte proliferative responses were also comparable. Thus, neither components of the candidate combination vaccine inhibited immune response to the other component. Substantial decrease of CHPV RNA and absence of histopathological changes in the brains of surviving immunized mice after challenge than the unimmunized controls further confirm efficacy of the vaccine even after intracerebral challenge. In conclusion, a combination vaccine seems feasible for use in a restricted area where the disease is endemic and should be subjected to additional studies required for future use in humans. PMID- 20005855 TI - Systematic review of interventions to increase influenza vaccination rates of those 60 years and older. AB - A systematic literature review identified 44 RCTs testing interventions to increase influenza vaccination rates among seniors >or=60. Case-control and cohort studies were excluded after review because of problems identifying secular trends and unknown confounders. Because of heterogeneity and unique interventions tested by a single or a few RCTs few studies could be pooled in meta-analysis. Using the CDC classification of interventions: (1) Increasing community demand: there is evidence of low quality that reminders increase influenza vaccination rates; (2) Increasing access: there is evidence of moderate quality that home visits to those >or=60 promoting influenza vaccination increase rates, and (3) Provider- and system-based interventions: there is evidence of moderate quality that facilitators working to improve preventive interventions in practices increase rates. PMID- 20005856 TI - Immunizing school-age children and adolescents: experience from low- and middle income countries. AB - OBJECTIVE: Given the increased attention on the need for booster immunizations of older children and adolescents, as well as new primary vaccine series that specifically target school-age children and adolescents, we reviewed the current state of vaccine delivery to school-age children and adolescents in low- and middle-income countries. METHODS: We searched the published literature and unpublished sources for articles, meeting presentations, technical reports and program documents related to immunization policies and programs for school-age children and/or adolescents between 6 and 19 years of age in low- and middle income countries. FINDINGS: We found several examples of ongoing school-age children and adolescent immunization in low- and middle-income countries. Reasons to vaccinate this age group include vaccines specifically targeted for this age group, waning immunity from prior vaccination, "catch-up" vaccination, acceleration of disease control or elimination efforts, and age distribution shift in the incidence of vaccine-preventable diseases. Multiple delivery strategies are currently in use: routine immunization, supplementary immunization activities, and Child Health Days and similar activities. Vaccines can be delivered in fixed sites, or through outreach. Most immunization programs that target adolescents and school-aged children are providing boosters of infant vaccines at school entry age, with scant experience in delivery of primary vaccination series in adolescents. Few of these programs have been formally evaluated and dissemination of lessons learned is limited. CONCLUSIONS: This baseline description may facilitate immunization program planning in countries considering vaccinating this age group. Additionally, this summary may inform plans for operational research and program evaluation designed to expand vaccine delivery to school-age children and adolescents in low- and middle-income countries. PMID- 20005857 TI - Affects of N-terminal variation in the SeM protein of Streptococcus equi on antibody and fibrinogen binding. AB - The clonal Streptococcus equi causes equine strangles, a highly contagious suppurative lymphadenopathy and rhinopharyngitis. An important virulence factor and vaccine component, the antiphagocytic fibrinogen binding SeM of S. equi is a surface anchored fibrillar protein. Two recent studies of N. American, Japanese and European isolates have revealed a high frequency of N-terminal amino acid variation in SeM of S. equi CF32 that suggests this region of the protein is subject to immunologic selection pressure. The aims of the present study were firstly to map regions of SeM reactive with convalescent equine IgG and IgA and stimulatory for lymph node cells and secondly to determine effects of N-terminal variation on the functionality of SeM. Variation did not significantly affect fibrinogen binding or susceptibility of S. equi to an opsonic equine serum. Linear epitopes reactive with convalescent IgG and mucosal IgA were concentrated toward the conserved center of SeM. However, IgA but not IgG from every horse reacted with at least one peptide that contained variable sequence. Lymph node cells (CD4+) from horses immunized with SeM were strongly responsive to a peptide (alphaalpha36-138) encoding the entire variable region. SeM (CF32) specific mouse Mab 04D11 which reacted strongly with this larger peptide but not with shorter peptides within that sequence reacted strongly with whole cells of S. equi CF32 but only weakly with cells of any of 14 isolates of S. equi expressing different variants of SeM. These results in combination suggest that N-terminal variation alters a conformational epitope of significance in mucosal IgA and systemic T cell responses but does not affect antibody mediated phagocytosis and killing. PMID- 20005858 TI - Cellular and humoral responses induced by Leishmania histone H2B and its divergent and conserved parts in cutaneous and visceral leishmaniasis patients, respectively. AB - Leishmania histone H2B has been reported to be a promising candidate for both vaccination and serodiagnosis. We evaluated the cellular immune responses induced by H2B and its divergent amino-terminal (H2B-N) and conserved carboxy-terminal (H2B-C) regions in individuals with a history of Localized Cutaneous Leishmaniasis (LCL) due to Leishmania (L.) major. H2B induced significantly high PBMC proliferation and IFNgamma levels in LCL individuals whereas significantly lower proliferation and IFNgamma levels were observed with the divergent part of the protein. All proteins induced IL10 in LCL and healthy individuals. We also evaluated the humoral responses induced by these proteins in patients with Mediterranean Visceral Leishmaniasis (MVL) due to L. infantum. H2B and H2B-N were highly recognized by MVL sera. Our results show that the entire H2B protein is more efficient than its amino- and carboxy-terminal regions in inducing a dominant Th1 profile in cured LCL subjects and suggest that this protein may constitute a potential vaccine against leishmaniasis. Furthermore, H2B and H2B-N are interesting antigens for serodiagnosis of MVL. PMID- 20005859 TI - Screening and identification of dominant functional fragments of human epididymal protease inhibitor. AB - To lay a foundation for the further development of a novel contraceptive vaccine based on epididymal protease inhibitor (Eppin) protein, possible B-cell epitopes were predicted using bioinformatics. Overlapping KLH-coupled peptides corresponding to the 89-133 amino acid domain of Eppin protein were synthesized by the Fmoc method. The male mice were immunized with various peptides resulting in a gradual elevation of specific serum IgG antibodies until they peaked at the seventh week. Treatment with antiserum in immunized mice caused a significant decrease in the sperm-egg binding rate. Eventually, antifertility assays in vivo showed that, the pregnancy rate and the number of births per labor in experimental mice were significantly decreased. The decrease in the F5 and the F4 groups were more outstanding therefore these peptides might be the dominant functional fragments of Eppin protein and provide an experimental basis for the development of effective contraceptive vaccine based on Eppin protein. PMID- 20005860 TI - Awareness of and attitude towards human papillomavirus infection and vaccination for cervical cancer prevention among adult males and females in Korea: a nationwide interview survey. AB - We assessed adult male and female awareness and acceptance of human papillomavirus (HPV) infection and vaccination for cervical cancer prevention, as well as factors associated with willingness to be administered the HPV vaccine. A nationwide population-based interview survey was conducted in Korea. One thousand male and female adults were included with random sampling. Despite the fact that awareness of HPV infection (13.3%) and the preventive effect of HPV vaccination (8.6%) were low, willingness to vaccinate against HPV (55.0%) was relatively high, especially with regard to participants' daughters (77.0%). Those who were informed about HPV infection (adjusted odds ratio, aOR=2.5 and 95% confidence interval, CI=1.1-5.3 in males; aOR=2.5 and 95% CI=1.5-4.2 in females) or vaccination (aOR=2.9 and 95% CI=1.1-7.6 in males; aOR=2.9 and 95% CI=1.6-5.4 in females), or who perceived a susceptibility to HPV infection (aOR=3.6 and 95% CI=1.9-6.8 in males; aOR=2.8 and 95% CI=1.6-5.0 in females) were more accepting of vaccination than those who did not. Appropriate knowledge transfer, guidance from a health professional, and reductions in the cost of the vaccine are key issues in promoting awareness of HPV infection and vaccination for cervical cancer prevention. PMID- 20005861 TI - Characterization of binding properties of monoglyceride lipase inhibitors by a versatile fluorescence-based technique. AB - Monoglyceride lipase (MGL) is a serine hydrolase that terminates the signaling of the primary endocannabinoid, 2-arachidonoyl glycerol (2-AG). Versatile high throughput screening methods allowing the testing of MGL inhibitors are rare, thereby limiting the development and analysis of novel inhibitors. Here we describe an improved fluorescence-based technique that is capable of determining time- and dose-dependent inhibition of MGL with one or multiple binding sites and, at the same time, is capable of revealing the reversibility of inhibitor binding in a simple kinetic assay format. Known reference compounds as well as novel inhibitors, such as JZL184 and CAY10499, were evaluated for their MGL binding properties and potency. PMID- 20005862 TI - Selection of appropriate reference genes for gene expression studies by quantitative real-time polymerase chain reaction in cucumber. AB - Quantitative real-time polymerase chain reaction (QRT-PCR) has become one of the most widely used methods for gene expression analysis. However, the expression profile of a target gene may be misinterpreted due to unstable expression of the reference genes under different experimental conditions. Thus, a systematic evaluation of these reference genes is necessary before experiments are performed. In this study, 10 putative reference genes were chosen for identifying expression stability using geNorm, NormFinder, and BestKeeper statistical algorithms in 12 different cucumber sample pools, including those from different plant tissues and from plants treated with hormones and abiotic stresses. EF1alpha and UBI-ep exhibited the most stable expression across all of the tested cucumber samples. In different tissues, in addition to expression of EF1alpha and UBI-ep, the expression of TUA was also stable and was considered as an appropriate reference gene. Evaluation of samples treated with different hormones revealed that TUA and UBI-ep were the most stably expressed genes. However, for abiotic stress treatments, only EF1alpha showed a relatively stable expression level. In conclusion, TUA, UBI-ep, and EF1alpha will be particularly helpful for reliable QRT-PCR data normalization in these types of samples. This study also provides guidelines for selecting different reference genes under different conditions. PMID- 20005863 TI - Protein detection by Western blot via coiled-coil interactions. AB - We propose an approach for the detection of proteins by Western blot that takes advantage of the high-affinity interaction occurring between two de novo designed peptides, the E and K coils. As a model system, K coil-tagged epidermal growth factor (EGF) was revealed with secreted alkaline phosphatase (SeAP) tagged with E coil (SeAP-Ecoil) as well as with biotinylated E coil. In that respect, we first produced purified SeAP-Ecoil and verified its ability to interact with K coil peptides by surface plasmon resonance biosensing. We demonstrated that protein detection with Ecoil-biotin was more specific than with SeAP-Ecoil. We then showed that our approach is as sensitive as conventional detection strategies relying on nickel-nitrilotriacetic acid-horseradish peroxidase (Ni-NTA-HRP), anti His-HRP, or anti-EGF. Altogether, our results indicate that the E/K coiled-coil system is a good alternative for protein detection by Western blot. PMID- 20005864 TI - Investigation of the chloride effect on hemoglobin by adsorptive transfer voltammetry. AB - A strategy of ex situ electrochemical method has been proposed for investigating the chloride effect on hemoglobin (Hb). Unlike the common electrochemical method that measures the chloride effect on Hb in bulk solution (in situ), the effects of chloride anion on Hb were investigated ex situ by adsorptive transfer voltammetry (AdTV) in this work. Gold electrode modified by self-assembled monolayer of 3-mercaptopropanoic acid (AuE/MPA) was prepared and then incubated in a series of Hb solutions containing different concentrations of chloride anion for adsorbing Hb-Cl (AuE/MPA/Hb-Cl). The resulting electrode was then measured in phosphate buffer solution by cyclic voltammetry. The corresponding voltammograms showed obvious promotion of the direct electron transfer of Hb with remarkable increase of peak currents, decrease of peak-to-peak separations, and negative shift of the formal potentials. As complementation, the adsorption behavior of Hb Cl on AuE/MPA, the structural information of Hb-Cl, and the electrocatalytic ability of AuE/MPA/Hb-Cl toward hydrogen peroxide were investigated by surface plasmon resonance, circular dichroism spectrum, ultraviolet-visible spectrum and amperometry, respectively. The results indicate that the chloride effect resulted in more electroactive sites of Hb on the surface of electrode. Meanwhile, the specific and nonspecific interactions between Hb and chloride anion can be discriminated from the electrochemical parameters obtained by AdTV. PMID- 20005865 TI - Heme, as a chaperone, binds to amyloid fibrils and forms peroxidase in vitro: possible evidence on critical role of non-specific peroxidase activity in neurodegenerative disease onset/progression using the alpha-crystallin-based experimental system. AB - We report that heme not only displays high binding affinity to the aggregates of crystallin, but also it is effectively able to interfere with this type of aggregation. In the present study, the influence of heme concentration on the crystallin fibrillogenesis was also investigated and experimental evidence of heme's prevention of crystallin aggregation was provided with the help of spectroscopic measurements. Significantly, using alpha-crystallin-based experimental system, we proposed that elevated levels of peroxidase activity may have a determinant role in amyloid pathogenesis. The substantial peroxidase activity of "crystallin aggregate-heme" may partially explain the acceleration of oxidative damage in several amyloid-affected neurodegenerative diseases. The present study also suggests that lipid peroxidation accompanying amyloidogenesis may be considered as a major cause in the pathogenesis of amyloid disorders. Since the consequence of heme-amyloid interaction has yet to be identified, additional data on it may help us to manage amyloid aggregation processes. PMID- 20005867 TI - Increased levels and defective glycosylation of MRPs in ovarian carcinoma cells resistant to oxaliplatin. AB - Pt compounds still represent the mainstay of the treatment of ovarian carcinoma. The aim of the present study was to investigate the molecular bases of resistance to Pt drugs using an oxaliplatin-resistant ovarian carcinoma cell model IGROV 1/OHP. These cells exhibited high levels of resistance to oxaliplatin, cross resistance to cisplatin and topotecan and displayed a marked accumulation defect of Pt drugs. This feature was associated with increased expression and altered N linked glycosylation of ATP binding cassette transporters MRP1 and MRP4. Pre treatment with tunicamycin, which inhibits the biosynthesis of N-linked oligosaccharides, decreased the accumulation of Pt in sensitive cells exposed to oxaliplatin or cisplatin and increased the electrophoretic mobility of MRP1 and MRP4, reproducing the association between decreased glycosylation of MRP1 and MRP4 and decreased Pt accumulation observed in the resistant IGROV-1/OHP cells. The observed N-glycosylation defect of oxaliplatin-resistant cells was linked to reduced levels of N-acetylglucosamine-1-phosphotransferase (GNPTG) and mannosyl (alpha-1,6-)-glycoprotein beta-1,6-N-acetyl-glucosaminyltransferase (MGAT5). This feature, observed in IGROV-1/OHP cells, was associated with decreased retention of Pt drugs. In addition, the overexpression of fully glycosylated MRP1 or MRP4 in tumor cell line of ovarian origin was associated with resistance to oxaliplatin and cisplatin. Our findings, showing that development of resistance to oxaliplatin results in up-regulation of MRPs, support that patients with oxaliplatin-refractory ovarian carcinomas may benefit from non-Pt-based regimens which do not contain MRP1 and MRP4 substrates. PMID- 20005866 TI - Assembly of the Sos1-Grb2-Gab1 ternary signaling complex is under allosteric control. AB - Allostery has evolved as a form of local communication between interacting protein partners allowing them to quickly sense changes in their immediate vicinity in response to external cues. Herein, using isothermal titration calorimetry (ITC) in conjunction with circular dichroism (CD) and macromolecular modeling (MM), we show that the binding of Grb2 adaptor--a key signaling molecule involved in the activation of Ras GTPase--to its downstream partners Sos1 guanine nucleotide exchange factor and Gab1 docker is under tight allosteric regulation. Specifically, our findings reveal that the binding of one molecule of Sos1 to the nSH3 domain allosterically induces a conformational change within Grb2 such that the loading of a second molecule of Sos1 onto the cSH3 domain is blocked and, in so doing, allows Gab1 access to the cSH3 domain in an exclusively non-competitive manner to generate the Sos1-Grb2-Gab1 ternary signaling complex. PMID- 20005868 TI - Effects of glyoxal or methylglyoxal on the metabolism of amino acids, lactate, glucose and acetate in the cerebral cortex of young and adult rats. AB - The in vitro effects of glyoxal and methylglyoxal on the metabolism of glycine, alanine, leucine, glutamate, glutamine, glucose, lactate and acetate were evaluated in cortico-cerebral slices from young (10-day-old) or adult (3-month old) rats. In a first set of experiments with cortico-cerebral slices from young animals, the compounds glyoxal or methylglyoxal at 400 microM, increased the oxidation of alanine, leucine and glycine to CO(2) and decreased the protein synthesis from these amino acids. Lipid synthesis from alanine, leucine and glycine was not changed in the cortico-cerebral slices from young rats after glyoxals exposure. Moreover, glutamine oxidation to CO(2) decreased by glyoxals exposure, but glutamate oxidation was not affected. In a second set of experiments with brain slices from adult animals, glycine metabolism (oxidation to CO(2), conversion to lipids or incorporation into proteins) was not changed by glyoxals exposure. In addition, the oxidation rates of glucose, lactate, acetate, glutamine and glutamate to CO(2) were also not modified. Taken together, these results indicate that glyoxal disrupts the energetic metabolism of the rat cerebral cortex in vitro. However, only young animals were susceptible to such events, suggesting that the immature cerebral cortex is less capable of dealing with glyoxal than the mature one. PMID- 20005869 TI - Effects of age and treadmill exercise in chronic diabetic stages on neuroblast differentiation in a rat model of type 2 diabetes. AB - In the present study, we investigated the effects of type 2 diabetes and treadmill exercise in chronic diabetic stages on neuroblast differentiation using doublecortin (DCX) in the subgranular zone of the dentate gyrus (SZDG) in Zucker diabetic fatty (ZDF) rats. Four-, 12-, 20- and 30-week-old Zucker lean control (ZLC) and ZDF rats were used to elucidate age-dependent changes of DCX immunoreactive neuroblasts. DCX-immunoreactive neuroblasts were significantly decreased with age in the SZDG. This reduction was prominent in the age-matched ZDF rats compared to that in the ZLC rats. To investigate the effects of treadmill exercise, ZLC and ZDF rats at 23 weeks of age were put on the treadmill with or without running for 1 h/day/5 consecutive days at 12-16 m/min for 7 weeks. Treadmill exercise significantly increased the tertiary dendrites of DCX immunoreactive neuroblasts in both ZLC and ZDF rats. In addition, exercise significantly increased the number of DCX-immunoreactive neuroblasts in the ZLC rats, but not in the ZDF rats. These results suggest that diabetes significantly decreases neuroblast differentiation and treadmill exercise in chronic diabetic animals has limitation to increase neuroblast differentiation although it increases neural plasticity. PMID- 20005870 TI - C. elegans BED domain transcription factor BED-3 controls lineage-specific cell proliferation during organogenesis. AB - The control of cell division is critical to organogenesis, but how this control is achieved is not fully understood. We found that mutations in bed-3, encoding a BED Zn-finger domain transcription factor, confer a phenotype where a specific set of cell divisions during vulval organogenesis is lost. Unlike general cell cycle regulators in Caenorhabditis elegans, the function of bed-3 is restricted to specific lineages. Transcriptional reporters suggest that bed-3 is expressed in a limited number of cell types including vulval cells whose divisions are affected in bed-3 mutants. A bed-3 mutation also affects the expression pattern of the cdh-3 cadherin gene in the vulva. The phenotype of bed-3 mutants is similar to the phenotype caused by mutations in cog-1 (Nkx6), a component of a gene regulatory network controlling cell type specific gene expression in the vulval lineage. These results suggest that bed-3 is a key component linking the gene regulatory network controlling cell-type specification to control of cell division during vulval organogenesis. PMID- 20005872 TI - Mesoderm and ectoderm lineages in the crustacean Parhyale hawaiensis display intra-germ layer compensation. AB - In Parhyale hawaiensis, the first three divisions are holoblastic and asymmetric, resulting in an embryo comprised of eight cells-four macromeres and four micromeres. Lineage studies performed at this stage demonstrate that the progeny of each cell contribute to specific portions of different germ layers. However, it is not known if this lineage pattern means a given blastomere is committed to its specific fate, indicative of mosaic development, or if regulation can occur between blastomere progeny so that the loss of a blastomere could be compensated for during development. Furthermore, if compensation occurs, what would be the source of such replacement? To investigate these possibilities, we performed ablation experiments at the eight-cell stage. We find that loss of blastomeres results in compensation. To determine the compensation pattern, we combined ablation and cell lineage tracing to reveal that progeny of mesoderm and ectoderm producing blastomeres display intra-germ layer compensation. Furthermore, by ablating lineages later in development, we identify a key interval between gastrulation and germband elongation after which compensation no longer occurs. Our results suggest that Parhyale possesses a mechanism to assess the status of mesoderm and ectoderm formation and alter development to replace the missing portions of these lineages. PMID- 20005871 TI - UNC-83 coordinates kinesin-1 and dynein activities at the nuclear envelope during nuclear migration. AB - Nuclei migrate during many events, including fertilization, establishment of polarity, differentiation, and cell division. The Caenorhabditis elegans KASH protein UNC-83 localizes to the outer nuclear membrane where it recruits kinesin 1 to provide the major motor activity required for nuclear migration in embryonic hyp7 cells. Here we show that UNC-83 also recruits two dynein-regulating complexes to the cytoplasmic face of the nucleus that play a regulatory role. One consists of the NudE homolog NUD-2 and the NudF/Lis1/Pac1 homolog LIS-1, and the other includes dynein light chain DLC-1, the BicaudalD homolog BICD-1, and the Egalitarian homologue EGAL-1. Genetic disruption of any member of these two complexes caused nuclear migration defects that were enhanced in some double mutant animals, suggesting that BICD-1 and EGAL-1 function in parallel to NUD-2. Dynein heavy chain mutant animals also had a nuclear migration defect, suggesting these complexes function through dynein. Deletion analysis indicated that independent domains of UNC-83 interact with kinesin and dynein. These data suggest a model where UNC-83 acts as the cargo-specific adaptor between the outer nuclear membrane and the microtubule motors kinesin-1 and dynein. Kinesin-1 functions as the major force generator during nuclear migration, while dynein is involved in regulation of bidirectional transport of the nucleus. PMID- 20005873 TI - Bone morphogenetic proteins mediate cellular response and, together with Noggin, regulate astrocyte differentiation after spinal cord injury. AB - Bone morphogenetic proteins (BMPs) play a critical role in regulating cell fate determination during central nervous system (CNS) development. In light of recent findings that BMP-2/4/7 expressions are upregulated after spinal cord injury, we hypothesized that the BMP signaling pathway is important in regulating cellular composition in the injured spinal cord. We found that BMP expressions were upregulated in neural stem cells (NSCs), neurons, oligodendrocytes and microglia/macrophages. Increased expression levels of pSmad1/5/8 (downstream molecules of BMP) were detected in neurons, NSCs, astrocytes, oligodendrocytes and oligodendroglial progenitor cells (OPCs). Active astrocytes which form the astroglial scar were probably derived from NSCs, OPCs and resident astrocytes. Since quiescent NSCs in the normal adult spinal cord will proliferate and differentiate actively into neural cells after traumatic injury, we proposed that BMPs can regulate cellular components by controlling NSC differentiation. Neurosphere culture from adult mouse spinal cord showed that BMP-4 promoted astrocyte differentiation from NSCs while suppressing production of neurons and oligodendrocytes. Conversely, inhibition of BMP-4 by Noggin notably decreased the ratio of astrocyte to neuron numbers. However, intrathecal administration of Noggin in the injured spinal cord failed to attenuate glial fibrillar acidic protein (GFAP) expression even though it effectively reduced pSmad expression. Noggin treatment did not block phosphorylation of Stat3 and the induction of GFAP in the injured spinal cord, suggesting that in addition to the BMP/Smad pathway, the JAK/STAT pathway may also be involved in the regulation of GFAP expression after spinal cord injury. PMID- 20005874 TI - Application of small interfering RNAs modified by unlocked nucleic acid (UNA) to inhibit the heart-pathogenic coxsackievirus B3. AB - This study describes the first application of unlocked nucleic acid (UNA) modified small interfering RNAs (siRNAs) directed against a medically relevant target, the coxsackievirus B3. We systematically analyzed the impact of different siRNA modification patterns and observed good compatibility of the introduction of UNA with the maintenance of high antiviral activity. Additionally, the polarity of an siRNA was successfully reversed by modulating the relative stability of the termini with locked nucleic acid (LNA) and UNA as shown in a reporter assay. The potency of the reversed siRNA against the full-length target was, however, too low to inhibit the infectious virus. Altogether, combined modification of siRNAs with LNA und UNA provides a promising approach to alter and improve properties of an siRNA. PMID- 20005875 TI - Dipeptide Tyr-Leu (YL) exhibits anxiolytic-like activity after oral administration via activating serotonin 5-HT1A, dopamine D1 and GABAA receptors in mice. AB - We found that Tyr-Leu (YL) dose-dependently exhibits potent anxiolytic-like activity (0.1-1mg/kg, i.p.) comparable to diazepam in the elevated plus-maze test in mice. YL was orally active (0.3-3mg/kg). A retro-sequence peptide or a mixture of Tyr and Leu was inactive. The anxiolytic-like activity of YL was inhibited by antagonists for serotonin 5-HT(1A), dopamine D(1) and GABA(A) receptors; however, YL had no affinity for them. We also determined the order of their activation is 5-HT(1A), D(1) and GABA(A) receptors using selective agonists and antagonists. Taken together, YL may exhibit anxiolytic-like activity via activation of 5 HT(1A), D(1) and GABA(A) receptors. PMID- 20005876 TI - Phylogenetically distinct Wolbachia gene and pseudogene sequences obtained from the African onchocerciasis vector Simulium squamosum. AB - Wolbachia are intracellular bacteria mostly found in a diverse range of arthropods and filarial nematodes. They have been classified into seven distinct 'supergroups' and other lineages on the basis of molecular phylogenetics. The arthropod-infecting Wolbachia are usually regarded as reproductive parasites because they manipulate their host species' sexing system to enhance their own spread, and this has led to their investigation as potential agents of genetic control in medical entomology. We report 12 partial Wolbachia gene sequences from: aspC, aspS, dnaA, fbpA, ftsZ, GroEL, hcpA, IDA, rpoB, rpe, TopI and wsp as well as a single ftsZ pseudogene sequence, which have all been PCR-amplified from Simulium squamosum (Diptera: Simuliidae). To our knowledge this is the first such report from Simuliidae. Uninterrupted open-reading frame sequences were obtained from all 12 genes, covering approximately 6.2kb of unique DNA sequence. Phylogenetic analyses with the different coding genes gave consistent results suggesting that the Wolbachia sequences obtained here do not derive from any of the known Wolbachia supergroups or lineages. Consistent with a unique genetic status for the S. squamosumWolbachia, the hypervariable regions of the Wolbachia specific wsp gene were distinct from all previous records in both sequence and length. As well as potential implications for newly emerging Wolbachia-based disease control methods, the results may be relevant to some problems experienced in the laboratory colonisation of Simulium damnosum sensu lato and why it is such a diverse species complex. PMID- 20005877 TI - Identification of functional FKB protein in Echinococcus granulosus: its involvement in the protoscolicidal action of rapamycin derivates and in calcium homeostasis. AB - FK506 (tacrolimus) and polyketide macrolides such as rapamycin and its derivates bind to FK506-binding proteins (FKBPs). These proteins display a peptidyl-prolyl rotamase function that is believed to catalyze protein folding and they are well validated anti-proliferative drug targets in certain pathogenic microorganisms, and their functions have been characterized in parasitic protozoa. However, much less is known in helminths and trials with rapalogs on cestoda have not yet been reported. Due to a growing need for new treatment options for human cystic echinococcosis, the in vitro efficacy of rapalogs in Echinococcus granulosus was investigated. We determined the effect of ramapycin, FK506 and everolimus against this cestode, demonstrating their protoscolicidal ability. Also, we observed synergic scolicidal actions during combined therapy with rapalogs plus cyclosporine A, proposing dual administration of drugs to improve pharmacological effects in vivo. We have identified an E. granulosus (Eg)-fkb1 gene that encodes Eg-FKBP, an archetypal protein of the FKBP family, which includes all residues implicated in the binding of pharmacological ligands, in the enzymatic activity and in interactions with possible target proteins. Levels of Eg-fkb1 mRNA are over-expressed by acid but not rapalog treatment. We also described the presence of receptor-operated calcium channels in the larval stage, suggesting that exogenous ligands may dissociate the interaction of Eg-FKBP from these intracellular channels, enhancing the activity of the Ca(2+) release and interfering with their normal regulatory functions. As rapamycin sensitivity is the major criterion used to detect targets of rapamycin kinase, we identified and analyzed in silico critical residues of putative homologs in the Echinococcus genome. These preliminary results will allow us to continue subsequent studies that could reveal the precise intracellular functions of Eg-FKBP, providing greater knowledge for further identification of downstream target proteins, a promising target for chemotherapy of cystic echinococcosis. PMID- 20005878 TI - Brain-independent development in the moth Sesamia nonagrioides. AB - The caterpillars of Sesamia nonagrioides developing under long-day (LD) photoperiod pupate in the 5th or 6th instar whereas under short day (SD) conditions they enter diapause and undergo several extra larval molts. The diapause is terminated within 1-3 instars upon transfer of SD larvae to the LD conditions. Brain removal from the 6th instar larvae promotes pupation followed by imaginal development; however, one third of the SD larvae and 12% of the LD larvae debrained at the start of the instar first undergo 1-2 larval molts. The incidence of larval molts is enhanced by the brain implants. Exclusively pupal molts occur in the LD larvae debrained late in the 6th instar. Decapitation elicits pupation in both LD and SD larvae, except for some of the 4th and 5th and rarely 6th instar that are induced to a fast larval molt. The pupation of decapitated larvae is reverted to a larval molt by application of a juvenile hormone (JH) agonist. No molts occur in abdomens isolated from the head and thorax prior to the wandering stage. Abdomens isolated later undergo a larval (SD insects) or a pupal (LD insects) molt. Taken together the data reveal that in S. nonagrioides (1) several larval molts followed by a pupal and imaginal molt can occur without brain; (2) an unknown head factor outside the brain is needed for the pupal-adult molt; (3) brain exerts both stimulatory and inhibitory effect on the corpora allata (CA); (4) larval molts induced in CA absence suggest considerable JH persistence. PMID- 20005879 TI - Loss of creatine in heart failure. A loss-win situation? PMID- 20005880 TI - On the role of diffusible binding partners in modulating the transport and concentration of proteins in tissues. AB - Here a reactive-diffusion transport model is used to demonstrate two previously undescribed functional roles for diffusible binding partners in the transport of molecules into tissues. The uptake of the insulin-like growth (IGF) and its binding partner the IGF binding protein (IGFBP3) into cartilage is used a specific tissue example to demonstrate a general principal. First, we show that reversible binding between free protein (IGF) and its diffusible binding partner (free IGFBPs) increases the rate of protein uptake into the tissue. Second, selective degradation of the binding partner can increase the transient and steady state free protein in tissues, well above the concentration at the source boundary, with the maximum free concentration occurring distant from the source boundary, deep within the tissue. This finding is very much at odds with expectations based on a traditional diffusion analysis. In cartilage, using realistic parameters, these new mechanisms raise the free IGF concentration by an order of magnitude deep within the tissue. As the increase in free protein is 'tunable' by cells, our analyses are postulated to demonstrate a general regulatory principle that may operate in any tissues throughout the body. PMID- 20005881 TI - Analysis of a stochastic SIR epidemic on a random network incorporating household structure. AB - This paper is concerned with a stochastic SIR (susceptible-->infective-->removed) model for the spread of an epidemic amongst a population of individuals, with a random network of social contacts, that is also partitioned into households. The behaviour of the model as the population size tends to infinity in an appropriate fashion is investigated. A threshold parameter which determines whether or not an epidemic with few initial infectives can become established and lead to a major outbreak is obtained, as are the probability that a major outbreak occurs and the expected proportion of the population that are ultimately infected by such an outbreak, together with methods for calculating these quantities. Monte Carlo simulations demonstrate that these asymptotic quantities accurately reflect the behaviour of finite populations, even for only moderately sized finite populations. The model is compared and contrasted with related models previously studied in the literature. The effects of the amount of clustering present in the overall population structure and the infectious period distribution on the outcomes of the model are also explored. PMID- 20005882 TI - Posterior parietal cortex mediates encoding and maintenance processes in change blindness. AB - It is commonly accepted that right posterior parietal cortex (PPC) plays an important role in updating spatial representations, directing visuospatial attention, and planning actions. However, recent studies suggest that right PPC may also be involved in processes that are more closely associated with our visual awareness as its activation level positively correlates with successful conscious change detection (Beck, D.M., Rees, G., Frith, C.D., & Lavie, N. (2001). Neural correlates of change detection and change blindness. Nature Neuroscience, 4, 645-650.). Furthermore, disruption of its activity increases the occurrences of change blindness, thus suggesting a causal role for right PPC in change detection (Beck, D.M., Muggleton, N., Walsh, V., & Lavie, N. (2006). Right parietal cortex plays a critical role in change blindness. Cerebral Cortex, 16, 712-717.). In the context of a 1-shot change detection paradigm, we applied transcranial magnetic stimulation (TMS) during different time intervals to elucidate the temporally precise involvement of PPC in change detection. While subjects attempted to detect changes between two image sets separated by a brief time interval, TMS was applied either during the presentation of picture 1 when subjects were encoding and maintaining information into visual short-term memory, or picture 2 when subjects were retrieving information relating to picture 1 and comparing it to picture 2. Our results show that change blindness occurred more often when TMS was applied during the viewing of picture 1, which implies that right PPC plays a crucial role in the processes of encoding and maintaining information in visual short-term memory. In addition, since our stimuli did not involve changes in spatial locations, our findings also support previous studies suggesting that PPC may be involved in the processes of encoding non-spatial visual information (Todd, J.J. & Marois, R. (2004). Capacity limit of visual short-term memory in human posterior parietal cortex. Nature, 428, 751-754.). PMID- 20005884 TI - Positive interactions, discontinuous transitions and species coexistence in plant communities. AB - The population and community level consequences of positive interactions between plants remain poorly explored. In this study we incorporate positive resource mediated interactions in classic resource competition theory and investigate the main consequences for plant population dynamics and species coexistence. We focus on plant communities for which water infiltration rates exhibit positive dependency on plant biomass and where plant responses can be improved by shading, particularly under water limiting conditions. We show that the effects of these two resource-mediated positive interactions are similar and additive. We predict that positive interactions shift the transition points between different species compositions along environmental gradients and that realized niche widths will expand or shrink. Furthermore, continuous transitions between different community compositions can become discontinuous and bistability or tristability can occur. Moreover, increased infiltration rates may give rise to a new potential coexistence mechanism that we call controlled facilitation. PMID- 20005883 TI - Septal co-infusions of glucose with the benzodiazepine agonist chlordiazepoxide impair memory, but co-infusions of glucose with the opiate morphine do not. AB - We have found repeatedly that medial septal (MS) infusions of glucose impair memory when co-infused with the gamma-amino butyric acid (GABA) agonist muscimol. The present experiments sought to determine whether the memory-impairing effects of this concentration of glucose would generalize to another GABA(A) receptor agonist and to an agonist from another neurotransmitter system that is known to impair memory. Specifically, we determined whether the dose of glucose that produces memory deficits when combined with muscimol in the MS would also impair memory when co-infused with the GABA(A) receptor modulator chlordiazepoxide (CDP) or the opiate morphine. Male Sprague-Dawley rats were given MS co-infusions and then 15 min later tested for spontaneous alternation or given shock avoidance training (retention tested 48 h later). The results showed that MS infusions of the higher dose of glucose with morphine did not produce memory deficits, whereas, the performance of rats given MS co-infusions of CDP with glucose was impaired. These findings suggest that the memory-impairing effects of brain glucose administration may involve an interaction with the GABA(A) receptor. PMID- 20005885 TI - The continuing evolution of torsades de pointes liability testing methods: is there an end in sight? AB - Drug-induced torsades de pointes (TdP) is a syndrome that includes a potentially lethal cardiac arrhythmia. It has been identified as a possible adverse drug reaction (ADR) for drugs which affect the repolarization processes of the heart. In order to predict the potential for TdP liability, regulatory guidelines have been developed which require that new drugs be safety screened. Unfortunately, however, despite this requirement there are no validated preclinical models with TdP incidence as a hard endpoint. Therefore, surrogate biomarkers are used. The most common and eliciting the most discussion/controversy among cardiovascular scientists is the duration of the QT interval of the ECG. Since no single model is available to wholly assess drug-induced TdP liability, safety pharmacologists employ a battery of complementary preclinical models in order to develop an integrated risk assessment (IRA). Ideally, the IRA should be comprised of the results from the effects of the new chemical entity (NCE) on the human ether-a-go go related (hERG) gene assay (actually a screen for block of the hERG gene product, the inward rectifying K current, IKr) and ECG effects in the conscious canine. However, since neither model is ideal the findings are generally supplemented by conduct of several additional experimental in vitro assays, namely the rabbit left ventricular wedge preparation, Langendorff isolated rabbit heart or isolated canine Purkinje fibre; nevertheless, as with many preclinical models, there is only limited validation and a resultant lack of general acceptance. Institution of regulatory guidance documents such as ICH S7A and S7B in conjunction with heightened awareness of the electrophysiological mechanisms that may be responsible for the development of TdP has led to a sharp fall in proarrhythmic compounds securing licensing, but at what costs? Supplementary experimental assays have furthered our understanding of drug-induced torsadogenesis, and it is now recognized that TdP is a multicausal event. This means that a perceived "positive" torsadogenic risk using one of the aforementioned models does not necessarily guarantee proarrhythmia. There has been an overall fall in the total number of NCEs pursued through development due to strict regulatory guidelines. Here we suggest that regulatory barriers can be alleviated by improving the integrated risk approach. But this requires better validation and deployment of existing preclinical models together with the invention of more precise and accurate models. PMID- 20005887 TI - Perchlorate transport and inhibition of the sodium iodide symporter measured with the yellow fluorescent protein variant YFP-H148Q/I152L. AB - Perchlorate is an environmental contaminant that impairs thyroid function by interacting with the sodium iodide symporter (NIS), the transporter responsible for iodide uptake in the thyroid gland. Perchlorate is well known as a competitive inhibitor of iodide transport by NIS, and recent evidence demonstrates that NIS can also transport perchlorate. In this study, we evaluated the yellow fluorescent protein (YFP) variant YFP-H148Q/I152L, as a genetically encodable biosensor of intracellular perchlorate concentration monitored by real time fluorescence microscopy. Fluorescence of recombinant YFP-H148Q/I152L was suppressed by perchlorate and iodide with similar affinities of 1.2 mM and 1.6 mM, respectively. Perchlorate suppressed YFP-H148Q/I152L fluorescence in FRTL-5 thyroid cells and NIS-expressing COS-7 cells, but had no effect on COS-7 cells lacking NIS. Fluorescence changes in FRTL-5 cells were Na+-dependent, consistent with the Na+-dependence of NIS activity. Perchlorate uptake in FRTL-5 cells resulted in 10-fold lower intracellular concentrations than iodide uptake, and was characterized by a higher affinity (K(m) 4.6 microM for perchlorate and 34.8 muM for iodide) and lower maximal velocity (V(max) 6.8 microM/s for perchlorate and 39.5 microM/s for iodide). Perchlorate also prevented iodide-induced changes in YFP-H148Q/I152L fluorescence in FRTL-5 cells, with half-maximal inhibition occurring at 1.1-1.6 muM. In conclusion, YFP-H148Q/I152L detects perchlorate accumulation by thyroid and other NIS-expressing cells, and reveals differences in the kinetics of perchlorate versus iodide transport by NIS. PMID- 20005888 TI - Safety assessment of personal care products/cosmetics and their ingredients. AB - We attempt to review the safety assessment of personal care products (PCP) and ingredients that are representative and pose complex safety issues. PCP are generally applied to human skin and mainly produce local exposure, although skin penetration or use in the oral cavity, on the face, lips, eyes and mucosa may also produce human systemic exposure. In the EU, US and Japan, the safety of PCP is regulated under cosmetic and/or drug regulations. Oxidative hair dyes contain arylamines, the most chemically reactive ingredients of PCP. Although arylamines have an allergic potential, taking into account the high number of consumers exposed, the incidence and prevalence of hair dye allergy appears to be low and stable. A recent (2001) epidemiology study suggested an association of oxidative hair dye use and increased bladder cancer risk in consumers, although this was not confirmed by subsequent or previous epidemiologic investigations. The results of genetic toxicity, carcinogenicity and reproductive toxicity studies suggest that modern hair dyes and their ingredients pose no genotoxic, carcinogenic or reproductive risk. Recent reports suggest that arylamines contained in oxidative hair dyes are N-acetylated in human or mammalian skin resulting in systemic exposure to traces of detoxified, i.e. non-genotoxic, metabolites, whereas human hepatocytes were unable to transform hair dye arylamines to potentially carcinogenic metabolites. An expert panel of the International Agency on Research of Cancer (IARC) concluded that there is no evidence for a causal association of hair dye exposure with an elevated cancer risk in consumers. Ultraviolet filters have important benefits by protecting the consumer against adverse effects of UV radiation; these substances undergo a stringent safety evaluation under current international regulations prior to their marketing. Concerns were also raised about the safety of solid nanoparticles in PCP, mainly TiO(2) and ZnO in sunscreens. However, current evidence suggests that these particles are non toxic, do not penetrate into or through normal or compromised human skin and, therefore, pose no risk to human health. The increasing use of natural plant ingredients in personal care products raised new safety issues that require novel approaches to their safety evaluation similar to those of plant-derived food ingredients. For example, the Threshold of Toxicological Concern (TTC) is a promising tool to assess the safety of substances present at trace levels as well as minor ingredients of plant-derived substances. The potential human systemic exposure to PCP ingredients is increasingly estimated on the basis of in vitro skin penetration data. However, new evidence suggests that the in vitro test may overestimate human systemic exposure to PCP ingredients due to the absence of metabolism in cadaver skin or misclassification of skin residues that, in vivo, remain in the stratum corneum or hair follicle openings, i.e. outside the living skin. Overall, today's safety assessment of PCP and their ingredients is not only based on science, but also on their respective regulatory status as well as other issues, such as the ethics of animal testing. Nevertheless, the record shows that today's PCP are safe and offer multiple benefits to quality of life and health of the consumer. In the interest of all stakeholders, consumers, regulatory bodies and producers, there is an urgent need for an international harmonization on the status and safety requirements of these products and their ingredients. PMID- 20005886 TI - PCB153-elicited hepatic responses in the immature, ovariectomized C57BL/6 mice: comparative toxicogenomic effects of dioxin and non-dioxin-like ligands. AB - Polychlorinated biphenyls (PCBs) are ubiquitous contaminants found as complex mixtures of coplanar and non-coplanar congeners. The hepatic temporal and dose dependent effects of the most abundant non-dioxin-like congener, 2,2',4,4',5,5' hexachlorobiphenyl (PCB153), were examined in immature, ovariectomized C57BL/6 mice, and compared to 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD), the prototypical aryl hydrocarbon receptor (AhR) ligand. Animals were gavaged once with 300 mg/kg PCB153 or sesame oil vehicle and sacrificed 4, 12, 24, 72 or 168 h post dose. In the dose-response study, mice were gavaged with 1, 3, 10, 30, 100 or 300 mg/kg PCB153 or sesame oil for 24 h. Significant increases in relative liver weights were induced with 300 mg/kg PCB153 between 24 and 168 h, accompanied by slight vacuolization and hepatocellular hypertrophy. The hepatic differential expression of 186 and 177 genes was detected using Agilent 4 x 44 K microarrays in the time course (|fold change|> or =1.5, P1(t)> or =0.999) and dose-response (|fold change|> or =1.5, P1(t)> or =0.985) studies, respectively. Comparative analysis with TCDD suggests that the differential gene expression elicited by PCB153 was not mediated by the AhR. Furthermore, constitutive androstane and pregnane X receptor (CAR/PXR) regulated genes including Cyp2b10, Cyp3a11, Ces2, Insig2 and Abcc3 were dose-dependently induced by PCB153. Collectively, these results suggest that the hepatocellular effects elicited by PCB153 are qualitatively and quantitatively different from TCDD and suggestive of CAR/PXR regulation. PMID- 20005889 TI - Orientation uncertainty reduces perceived obliquity. AB - The influence of prejudice on perception should be greatest when certainty about stimulus identity is least. We exploited this relationship to reveal visual biases for the cardinal orientations: vertical and horizontal. Specifically, when we increased the variance of orientations in an array of grating patches, estimates of the mean became less oblique. This result is consistent with a stable prior, or prejudice, for those orientations most prevalent in natural scenes. PMID- 20005891 TI - Sphingolipid metabolism in trans-golgi/endosomal membranes and the regulation of intracellular homeostatic processes in eukaryotic cells. PMID- 20005890 TI - Anti-SPARC oligopeptide inhibits laser-induced CNV in mice. AB - It is known that SPARC gates VEGF-A signal transduction towards KDR, the primary angiogenic VEGF receptor. We sought to determine whether inhibition of SPARC activity using anti-SPARC peptide could inhibit laser-induced CNV by promoting binding of VEGF-A to FLT-1. We created anti-SPARC l-peptide and retro-inverso anti-SPARC d-peptide. Anti-SPARC peptides or PBS were injected intravitreally 1day before or after laser induction. Intravitreal injection of anti-SPARC l peptide 1day before laser induction promotes FLT-1 phosphorylation and inhibited laser-induced CNV and anti-SPARC d-peptide had no effect. Injection 1day after laser injury did not affect size of laser-induced CNV. Inhibition of SPARC activity could be complementary to existing anti-CNV therapy. PMID- 20005892 TI - Structural basis for H3K4 trimethylation by yeast Set1/COMPASS. PMID- 20005893 TI - Regulation of cocaine self-administration in humans: lack of evidence for loading and maintenance phases. AB - BACKGROUND: In rodents, cocaine self-administration under a fixed-ratio schedule and with timeout intervals limited to the duration of the infusions is characterized by an initial burst of drug intake (loading) followed by more stable infusion rates (maintenance). We sought to examine whether similar phases might characterize self-regulated cocaine use in humans. METHODS: 31 Non treatment seeking, cocaine dependent subjects participated in three (8, 16, and 32 mg/70 kg/infusion), self-regulated, 2-h cocaine self-administration sessions under a fixed-ratio 1, 5-min timeout schedule. Data were assessed for visual (e.g., by graphs of cumulative numbers of infusions) and statistical evidence of change in phase (by step-function analyses of individual infusion rates). RESULTS: Graphs of cumulative infusions over time suggested a single, linear rate of self-administration over 2h at each cocaine dose. Statistical analyses of infusion data by generalized estimating equation (GEE) models also failed to support a loading/maintenance pattern (suggesting, if anything, the possibility of increasing infusion rates over time). CONCLUSIONS: Our findings fail to support the existence of distinct loading and maintenance phases of self regulated cocaine administration in humans at behaviorally relevant doses. Several factors may account for these observations including differences between humans and rodents in self-regulated drug intake. PMID- 20005894 TI - Glucocorticoid sensitivity of cognitive and inflammatory processes in depression and posttraumatic stress disorder. AB - Both hyper- and hypo-activity of the hypothalamus-pituitary-adrenal (HPA) axis activity are a consistently reported hallmark feature of stress-related disorders, such as major depression and posttraumatic stress disorder (PTSD), respectively. In this manuscript, however, we are summarizing evidence pointing to altered glucocorticoid (GC) sensitivity in relevant target tissues for HPA axis hormones. Specifically, we provide a summary of GC effects on cognitive functions, as an emerging marker for central nervous system GC sensitivity, and of GC effects on peripheral inflammatory responses. With regard to depression and PTSD, evidence thereby points to decreased GC sensitivity of the cognitive and inflammatory systems in depression, and increased GC sensitivity of both systems in PTSD. Taken together, these data support the hypothesis that both psychiatric disorders are characterized by inefficient GC signaling, although through dysregulations at different levels. Potential underlying pathways and implications are discussed. PMID- 20005895 TI - Analysis of novel phospho-ITAM specific antibodies in a S2 reconstitution system for TCR-CD3 signalling. AB - The T cell antigen receptor (TCR-CD3) complex contains 12 different cytoplasmic tyrosines, each of which is part of an immunoreceptor tyrosine-based activation motif and thus occurs in similar sequence context. Since phosphorylation of individual tyrosines can be correlated with the quality of the T cell response, monitoring their phosphorylation is important. We thus generated novel antibodies against phospho-tyrosines of the TCR-CD3 complex and tested the specificity in a synthetic biology approach. We utilized the Drosophila S2 reconstitution system testing several kinases and stimulation conditions that lead to optimal phosphorylation of the TCR-CD3 subunit zeta. Expressing TCR-CD3 subunits and tyrosine mutants thereof we tested the specificity of the novel antibodies in Western blot and immunopurification experiments. In particular, we generated and characterized the monoclonal antibody EM-26 that specifically recognizes phosphorylation of the membrane proximal tyrosine of zeta (phospho-zetaY1) and antisera raised against the first and the second phospho-tyrosine of CD3epsilon (phospho-epsilonY1 and phospho-epsilonY2). PMID- 20005896 TI - A closer look into the GL7 antigen: its spatio-temporally selective differential expression and localization in lymphoid cells and organs in human. AB - The GL7 epitope was originally described as part of a late lymphocyte activation antigen expressed in mouse and widely used since then as a marker of germinal center. Here we report on its differential expression by rat and human immune cells and lymphoid organs. Expression pattern of the GL7 epitope in rats is similar to that described earlier in mice, namely that GL7 antigen appears only on lymphocytes after 48h activation. In humans lymphocytes, but not the differentiated cells of myeloid origin, express this epitope. The GL7 epitope is up-regulated upon in vitro activation of primary T cells, while a slightly decreased expression is found on B lymphocytes. Fluorescent immunohistochemistry shows discrete location of GL7(hi) cells in human tonsil. GL7 antibody intensely stains CD19(+), IgD(+), IgM(low) B lymphocytes found at the margin of B cell follicles. The GL7 epitope is constitutively and highly raft-associated in human lymphoid cells. Strong neuraminidase- and partial papain-sensitivity of the GL7 epitope on human lymphocytes indicates a sialic acid-containing epitope linked either to one (or more) membrane protein(s) or to lipids. The lymphocyte restricted GL7 epitope expression and the activation-dependent bi-directional change in the amount of the epitope suggest a functional role for GL7 epitope linked to carbohydrate-based immunoregulation. PMID- 20005897 TI - Transgenic mice expressing human FcgammaRIIa have enhanced sensitivity to induced autoimmune arthritis as well as elevated Th17 cells. AB - The major human Fc receptor, huFcgammaRIIa, is implicated in the development of autoimmune arthritis in humans but until recently has not been studied in mouse models. We evaluated potential roles of FcgammaRIIa by using transgenic mice expressing the receptor. We examined two models of induced autoimmune arthritis pristane-induced arthritis (PIA) and collagen-induced arthritis (CIA) as well as the anti-collagen-II antibody-induced arthritis (CAIA) model. In the induced arthritis models PIA and CIA, the transgenic mice developed a more severe arthritis than the other arthritis-prone SJL or DBA1 mice. Interestingly, anti collagen-II antibodies were elevated in PIA in the susceptible mice. In the CIA model, the highly susceptible transgenic mouse had IgG subclass levels equivalent to the unaffected and disease resistant C57BL/6 mouse strain implying that the FcgammaRIIa lowers the threshold of IgG dependent leukocyte activation. This is consistent with the greatly enhanced sensitivity of the FcgammaRIIa transgenic mice to CAIA which clearly indicates a role for the receptor at least at the inflammatory effector cell level. Other roles for huFcgammaRIIa or other gene products in the development of autoimmunity cannot be ruled out however, especially as the mice exhibited elevated Th1 or Th17 CD4 T cells in the draining lymph nodes. PMID- 20005898 TI - Detection of phosphorylated T and B cell antigen receptor species by Phos-tag SDS and Blue Native-PAGE. AB - Detection of phospho-proteins and differently phosphorylated forms of the same protein are important in understanding cell behaviour. One novel method is Phos tag SDS-PAGE. A dinuclear Mn(2+) complex that binds to phosphate groups (the Phos tag) is covalently attached to the polyacrylamide gel matrix. Thus, phosphorylated proteins are retarded in their migration and can be distinguished from their non-phosphorylated counterparts. We applied Phos-tag SDS-PAGE to the analysis of the zeta, CD3epsilon and CD3delta subunits of the T cell antigen receptor (TCR-CD3). Pervanadate stimulation generated six different phospho-zeta and each two different CD3epsilon and CD3delta forms. This corresponds to the phosphorylatable tyrosines on their cytoplasmic tails. The phosphorylation pattern was compatible with random phosphorylation events. Further, we showed that the Phos-tag technology can be applied to Blue Native (BN)-PAGE. This extends the applicability to the analysis of native protein complexes. Upon pervanadate stimulation the TCR-CD3 complex was predominantly detected as two distinct phospho-complexes. In contrast, the B cell antigen receptor (BCR) appeared as one phospho-form. Thus, Phos-tag BN-PAGE is useful for the analysis of different phosphorylation states of multiprotein complexes. PMID- 20005899 TI - Signals and Signal Processing in the Immune System. Introduction. PMID- 20005900 TI - The role of dendritic cells in mycobacterium-induced granulomas. AB - The presence of dendritic cells (DCs) in mycobacterium-containing granulomas, as well as in other granuloma-inducing diseases, is beginning to be appreciated. This review will summarize what is known about DCs with regards to the granuloma and discuss the potential roles DCs may be playing during mycobacterial infection. Potential functions may include mycobacterial dissemination from lesions or sampling of granuloma-containing mycobacterial antigens and migration to the draining lymph nodes to maintain continuous T cell priming. Additionally, the review will discuss the potential outcomes of DC-T cell cross-talk within the granuloma and whether it results in boosting the effector functions of newly arrived T cells or anergizing systemic T cells locally. Understanding the DCs complex and changing role during this critical stage may help explain how latency is achieved and maintained. Such knowledge might also lead to improved vaccination strategies. PMID- 20005901 TI - Transglutaminase 2 is expressed and active on the surface of human monocyte derived dendritic cells and macrophages. AB - The multifunctional enzyme, transglutaminase 2 (TG2), can be found intracellularly, in the extracellular matrix and on the cell surface. Cell surface TG2 (csTG2) could not be detected by TG2-specific antibodies or autoantibodies on immunocompetent cells. A supposedly csTG2-specific antibody, 6B9, was recently shown to actually react with CD44. Though the importance of TG2 mediated deamidation of gluten in the pathogenesis of celiac disease has been well recognized, it is not known in which intestinal cells or cell compartment the deamidation occurs. Duodenal dendritic cells (DCs) can be directly involved in gluten-reactive T-cell activation. Here we use blood monocyte-derived dendritic cells (iDC) and macrophages (MPhi) as a model for intestinal antigen presenting cells (APCs) and show that they contain large amounts of TG2. We found that TG100, a commercial TG2-specific monoclonal antibody can recognize TG2 on the surface of these cells, that is monocyte-derived APCs express surface associated TG2. TG2 expression was found on the surface of individual tunica propria cells in frozen small bowel tissue sections from both normal and celiac subjects. We also demonstrate that the pool of TG2 on the surface of iDCs can be catalytically active, hence it might directly be involved in the deamidation of gliadin peptides. Bacterial lipopolysaccharide (LPS) increased the level of TG2 on the surface of maturing DCs, supporting the hypothesis that an unspecific inflammatory process in the gut may expose more transglutaminase activity. PMID- 20005902 TI - Chemokine-chemokine receptor axes in melanoma brain metastasis. AB - Brain metastasis confers an extremely unfavorable prognosis upon melanoma patients. The mechanisms underlying the homing of metastatic melanoma to the brain and survival of metastatic melanoma cells in the brain are unknown. Tumor cells, including melanoma, use chemokine receptor-ligand axes to home to specific organ sites. To identify chemokine receptors that might be involved in brain targeted melanoma metastasis, we first established a chemokine receptor profile of cultured melanoma cells (3 cell lines of cutaneous melanoma and 5 cell lines of melanoma brain metastasis). The expression of the membrane-bound chemokine CX3CL1 by these lines was also determined. We show that out of 19 receptors tested, cultured melanoma cells express CCR3, CCR4, CXCR3, CXCR7, CX3CR1 and membrane CX3CL1. Utilizing cells from newly created variants of human melanoma xenografts, we found that the expression of CCR4 was significantly higher in one brain metastatic variant compared to its expression in the corresponding local variant. Local and metastatic variants stimulated with the CCR4 ligand, CCL22, showed a differential AKT phosphorylation pattern. These findings may suggest the involvement of CCR4 in the process of brain metastasis in human melanoma, and that CCR4 may be a novel molecular biomarker for the identification of melanoma cells likely to metastasize to the brain. PMID- 20005903 TI - Rapid method for SV40 titration. AB - SV40 titer is determined traditionally by the conventional plaque assay. Plaques appear after several rounds of infection and the assay takes around two weeks, which may delay research. A simpler assay was developed, based on detection of T antigen in the infected cells by flow cytometry. Cells grown in 6-well plates are infected with serial dilutions of the viral stock, harvested 48h post-infection, stained and analyzed for T-antigen using a flow cytometer. The viral titer is calculated based on the percentage of T-antigen positive cells. The procedure is accomplished in 2 days. Unexpectedly we found that titers on different permissive African Green Monkey kidney cell lines were consistently different, suggesting variable susceptibility to SV40 infection. The method described, optimized for SV40 titration, may be adapted readily to other viruses. PMID- 20005904 TI - One-step real-time RT-PCR for pandemic influenza A virus (H1N1) 2009 matrix gene detection in swine samples. AB - In the spring of 2009, a novel (H1N1) influenza A virus began to spread among humans worldwide. Although the 2009 H1N1 is related genetically to swine influenza viruses, human infection has not been connected to pig exposure. Because the virus is now circulating widely in the human population, swine herds are at increased risk of becoming infected. In order to investigate potential outbreaks of the 2009 pandemic virus in pigs, a quantitative real-time reverse transcriptase-polymerase chain reaction (qRT-PCR) for the detection of the (H1N1) 2009 RNA in clinical specimens was developed. To evaluate the applicability of the test as a diagnostic tool in the screening of field specimens from swine, 64 field isolates of North American swine, 5 equine and 48 avian influenza viruses collected during diagnostic investigations were analyzed retrospectively as well as samples collected during an experimental in vivo infection with two novel H1N1 isolates, A/California/04/2009 (H1N1)v virus and A/Mexico/4108/2009 (H1N1)v. The sensitivity of the qRT-PCR was shown to be higher with respect to standard techniques such as virus isolation and the reproducibility was satisfactory. The present unique and highly sensitive assay is able to detect as little as 1 x 10(1) copies of RNA per microl of template and it represents a rapid and useful approach for the screening and quantitation of (H1N1) 2009 RNA in porcine specimens. PMID- 20005905 TI - RNA immunization can protect mice against foot-and-mouth disease virus. AB - In previous work we have reported the immunization of swine using in vitro transcribed foot-and-mouth disease virus (FMDV) RNA. With the aim of testing whether RNA-induced immunization can mediate protection against viral infection, a group of Swiss adult mice was inoculated with FMDV infectious transcripts. In most inoculated animals viral RNA was detected in serum at 48-72h postinoculation. A group of the RNA-inoculated mice (11 out of 19) developed significant titers of neutralizing antibodies against FMDV. Among those animals that were successfully challenged with infectious virus (15 out of 19), three out of the eight animals immunized upon RNA inoculation were protected, as infectious virus could not be isolated from sera but specific anti-FMDV antibodies could be readily detected. These results suggest the potential of the inoculation of genetically engineered FMDV RNA for virulence and protection assays in the murine model and allow to explore the suitability of RNA-based FMDV vaccination in natural host animals. PMID- 20005906 TI - Endothelin-1 plasma concentrations in patients with retinitis pigmentosa. AB - PURPOSE: The aim of this study was to investigate the behaviour of plasma endothelin-1 (ET-1) levels in patients affected by retinitis pigmentosa (RP) and syndromic RP. METHODS: Blood samples were obtained from a group of 40 consecutive patients with RP matched with 35 healthy subjects (HS) as control. We carried out a complete ophthalmological examination. The study group included 26 patients with RP and 14 patients with syndromic RP. Plasma ET-1 levels were determined in duplicate with a specific radioimmunoassay method. RESULTS: In the HS plasma ET-1 levels were 7.48+/-2.58 pg/mL. The mean of plasma ET-1 concentrations in all patients with RP ( 16.2+/-5.6 pg/mL) was significantly (P<0.01) higher than that of HS. Moreover, in the syndromic RP patients, plasma ET-1 levels (18.9+/-6.8 pg/mL) were higher than those of HS and RP patients (P<0.01). CONCLUSION: The increase of plasma ET-1 levels in RP patients suggests that ET-1 may play a role in the pathophysiology of the diseases involving retinal pigment epithelial cells and the retinal vascular system such as RP. PMID- 20005907 TI - Cellular distribution of chromogranin A in excitatory, inhibitory, aminergic and peptidergic neurons of the rodent central nervous system. AB - Immunoreactivity for both processed and unprocessed forms of chromogranin A (CGA) was examined, using an antibody recognizing the WE14 epitope, among terminal fields and cell bodies of anatomically defined GABAergic, glutamatergic, cholinergic, catecholaminergic, and peptidergic cell groups in the rodent central nervous system. CGA is ubiquitous within neuronal cell bodies, with no obvious anatomical or chemically-coded subdivision of the nervous system in which CGA is not expressed in most neurons. CGA expression is essentially absent from catecholaminergic terminal fields in the CNS, suggesting a relative paucity of large dense-core vesicles in CNS compared to peripheral catecholaminergic neurons. Extensive synaptic co-localization with classical transmitter markers is not observed even in areas such as amygdala, where CGA fibers are numerous, suggesting preferential segregation of CGA to peptidergic terminals in CNS. Localization of CGA in dendrites in some areas of CNS may indicate its involvement in regulation of dendritic release mechanisms. Finally, the ubiquitous presence of CGA in neuronal cell somata, especially pronounced in GABAergic neurons, suggests a second non-secretory vesicle-associated function for CGA in CNS. We propose that CGA may function in the CNS as a prohormone and granulogenic factor in some terminal fields, but also possesses as-yet unknown unique cellular functions within neuronal somata and dendrites. PMID- 20005908 TI - PTH inactivates the AKT survival pathway in the colonic cell line Caco-2. AB - In previous works, we found that PTH promotes the apoptosis of human Caco-2 intestinal cells, through the mitochondrial pathway. This study was conducted to investigate the modulation of different players implicated in the AKT survival pathway in PTH-induced intestinal cell apoptosis. We demonstrate, for the first time, that PTH modulates AKT phosphorylation in response to apoptosis via the serine/threonine phosphatase PP2A. PTH treatment induces an association of AKT with the catalytic subunit of PP2A and increases its phosphatase activity. PTH also promotes the translocation of PP2Ac from the cytosol to the mitochondria. Furthermore, our results suggest that PP2A plays a role in hormone-dependent Caco 2 cells viability and in the cleavage of caspase-3 and its substrate PARP. The cAMP pathway also contributes to PTH-mediated AKT dephosphorylation while PKC and p38 MAPK do not participate in this event. Finally, we show that PTH induces the dissociation between 14-3-3 and AKT, but the significance of this response remains unknown. In correlation with PTH-induced Bad dephosphorylation, the hormone also decreases the basal association of 14-3-3 and Bad. Overall, our data suggest that in Caco-2 cells, PP2A and the cAMP pathway act in concert to inactivate the AKT survival pathway in PTH-induced intestinal cell apoptosis. PMID- 20005909 TI - Model based optimization of feeding regimens in aquaculture: application to the improvement of Octopus vulgaris viability in captivity. AB - The culture of common octopus (Octopus vulgaris), one important candidate to the aquaculture diversification, faces significant difficulties, mainly related with an inadequate first development stages diet. A mathematical model integrating disperse information on the nutrient composition throughout the species ontogenic development as well as on the effects of broodstock feeding and diet composition data of O. vulgaris, allowed us to predict the time evolution of paralarvae nutritional composition in terms of protein and lipid fractions and to design an optimal diet composition with the objective to ensure the maximal survival. The optimization routine showed that a diet based on the spider crab (Maja squinado) zoea composition is the most suitable for reaching the best survival rates. Results are verified by comparison with available experimental data. The obtained results and the prospective developments are a good example of how the systemic, quantitative model based approach can be used to analyse and contribute to the understanding of complex biological systems. PMID- 20005910 TI - Hepatitis B virus overexpresses suppressor of cytokine signaling-3 (SOCS3) thereby contributing to severity of inflammation in the liver. AB - The mechanism by which hepatitis B virus (HBV) infection causes severe inflammatory liver diseases is multifactorial and related to interactions with cell signaling pathways and the ensuing inflammatory response. Activation of JAK/STAT/SOCS signaling is essential for the induction of cellular antiviral responses, contributes to apoptosis and is negatively regulated by SOCS proteins. Recent reports have shown that SOCS3 activation interferes with viral protein expression and treatment response and thereby plays a major role in hepatitis virus infections. We analyzed the expression of SOCS3 in liver specimens from HBV infected patients using immunohistochemistry (IHC) and determined the effect of HBV on STAT/SOCS signaling in functional cell culture experiments (HuH-7) using HBV-expressing adenoviral constructs (AdHBV). Increased expression of SOCS3 protein was identified in liver specimens from patients with chronic HBV infection and this correlated with the severity of liver inflammation. In accordance with the IHC-findings, in vitro analyses demonstrated that HBV infection of HuH7 cells was associated with increased expression of SOCS3 protein. In spite of the over expression of its negative regulator SOCS3 we observed a constitutive activation of STAT3. SOCS1 levels were not increased while pSTAT1 was suppressed in HBV-infected HuH7 cells. Our results demonstrate that STAT/SOCS-signaling is dysregulated in HBV-infected hepatocytes both in vivo and in vitro and this correlated with the severity of liver inflammatory changes. This interference of STAT/SOCS signaling by HBV may result in an ineffective immune response against HBV and potentially contributes to viral pathogenesis, malignant transformation and may represent an important mechanism of viral persistence. PMID- 20005911 TI - HPMA copolymers for masking and retargeting of therapeutic viruses. AB - Hydrophilic polymers are widely used already for steric stabilisation of bioactive proteins, changing their pharmacokinetics and modifying their interactions with the biological environment. Polymers may also be conjugated to biological surfaces, such as viruses, bacteria and mammalian cells, also to endow steric protection and changed properties. Reactive polymers based on N-[2 hydroxypropyl]methacrylamide have shown particular promise for surface coating of viruses, particularly adenovirus, and here we describe the important observations and innovations arising from this combination of chemical and genetic engineering. Adenovirus is a versatile agent that already finds important experimental applications as a recombinant vaccine, and also for cancer therapy, although its activity in both settings is often limited by a potent antibody neutralising response in humans that is generally not seen in experimental animals. Coating with HPMA copolymers provides protection against neutralisation by antibodies and complement, and covalent linkage of novel ligands to the surface of the polymer can endow new infectious tropisms, mediated through different receptors, that can expand the potential applications of this versatile technology for a range of settings. PMID- 20005912 TI - Salt preference of nursing mothers is associated with earlier cessation of exclusive breastfeeding. AB - Successful breastfeeding is predicated on its initial success. Salt appetite during lactation may be relevant to breastfeeding success because sodium is essential for development of foetus and neonate. Here we examined whether maternal salt preference might facilitate breastfeeding. Nursing mothers (n=327) were categorized as high, medium or low salt preferring, and the relationship to persistence of exclusive breastfeeding during the first 25 days postnatal was evaluated. Contrary to expectation, we find that mothers with low salt preference persisted in breastfeeding beyond day 7 postnatal in comparison to mothers with high salt preference, and mothers with high salt preference had the shortest exclusive breastfeeding duration up to postnatal day 25. Awareness of this among health workers and nursing mothers could contribute to successful breastfeeding. PMID- 20005913 TI - The withdrawal from TDF therapy could induce transient RLS. PMID- 20005914 TI - Estimation of dietary intake of 5-hydroxymethylfurfural and related substances from coffee to Spanish population. AB - 5-Hydroxymethylfurfural (HMF) is naturally formed during food processing or cooking activities, giving its ubiquity in the Western diet. HMF could be metabolised to 5-sulfooxymethylfurfural making HMF potentially harmful in an extent unknown at present. Coffee is the main exposure source. Occurrence of HMF, 5-hydroxymethyl-2-furoic acid (HMFA) and 2-furoic acid (FA) were measured in commercial ground coffee and soluble coffee marketed in Spain. Levels of 110, 625, 1734, 2480 mg HMF/kg were obtained for natural, blend, torrefacto and soluble coffee, respectively, giving four classes significantly different. Soluble coffee showed the largest variability in HMF. Levels of HMFA and FA did not change significantly being about 600 mg/kg. Dietary exposure to HMF coffee to consumption in the total Spanish population was estimated to be 8.57 mg/day by using a deterministic approach. However, median level was recalculated to 5.26 mg HMF/day when specific contribution of each type of ground and soluble coffee in the consumption habits was considered. Resultant value is above of the threshold of concern (1600 microg HMF/day, mTAMDI). A level of 8.57 mg HMF/day in persons with high consumption habits (95th percentile) was calculated for risk assessment. PMID- 20005915 TI - The influence of age on tooth supported fixed prosthetic restoration longevity. A systematic review. AB - OBJECTIVE: The purpose of this study was to investigate the possible influence of age on the longevity of tooth supported fixed prosthetic restorations, using a systematic review process. DATA SOURCES: To identify relevant papers an electronic search was made using various databases (MEDLINE via Pubmed, EMBASE, The Cochrane Register of RCTs, the database of abstracts of Reviews of Effects DARE), augmented by hand searching of key prosthodontic journals (International Journal of Prosthodontics, Journal of Prosthetic Dentistry and Journal of Prosthodontics) and reference cross-check. STUDY SELECTION: Assessment and selection of studies identified were conducted in a two phase procedure, by two independent reviewers utilizing specific inclusion and exclusion criteria. The minimum mean follow-up time was set at 5 years. RESULTS: The initial database search yielded 513 relevant titles. After the subsequent filtering process, 22 articles were selected for full-text analysis, finally resulting in 11 studies that met the inclusion criteria. All studies were classified as category C according to the strength of evidence. Meta-analysis was not possible due to the non-uniformity of the data available. The final studies were presented with conflicting results. The majority of the final studies did not report a statistically significant effect of age on fixed prostheses survival, whilst only one study reported poorer prognosis for elderly patients, and two studies reported poorer prognosis for middle-aged patients. CONCLUSIONS: The results of this systematic review showed that increased age of patients should not be considered as a risk factor for the survival of fixed prostheses. Although the majority of studies did not show any effect of age on the survival of fixed prostheses, there was some evidence that middle-aged patients may present with higher failure rates. PMID- 20005916 TI - Usage of signaling in neurodegeneration and regeneration of peripheral nerves by leprosy bacteria. AB - Multiple signaling pathways play key regulatory roles during the development of peripheral nervous system (PNS) and also in neuroregeneration process following nerve degeneration. Schwann cells, the glial cells of the PNS, by interacting with neuronal (axonal) ligands, mainly neuregulins via receptor tyrosine kinase (RTK) complex, ErbB2/ErbB3, initiate intracellular signaling pathways to drive proliferation and differentiation of Schwann cells, both during development and the process of regeneration and re-myelination after nerve injury. One of the major signaling kinases, extracellular signal-regulated kinase-1/2 (ERK1/2), that is also a downstream signaling pathway of neuregulin-ErbB2/ErbB3 activation, has been identified as a key regulator of Schwann cell proliferation, differentiation, demyelination and nerve regeneration. Recent studies have provided evidence that the bacterium that causes human leprosy, Mycobacterium leprae that has a unique capacity to invade Schwann cells of the adult PNS, utilizes the neuregulin-ErbB2/ErbB3 associated signaling network to the bacterial advantage. M. leprae directly bind to ErbB2 on myelinated Schwann cells and activate the RTK by a novel route that bypasses the classical neuregulin/growth factor-induced ErbB2-ErbB3 heterodimerization, and subsequently induce downstream the canonical Erk1/2 signaling, leading to myelin breakdown and subsequent axonal damage. This initial injury provides a survival advantage for M. leprae as it induces de-differentiation and generates myelin-free cells, which are highly susceptible to M. leprae invasion and promote bacterial survival. Once invaded M. leprae activate Erk1/2 via a non-canonical pathway and subsequently increase the cell proliferation and maintain the infected cells in de-differentiated state, thereby preventing remyelination. Therefore, by subverting major RTKs and signaling pathways in adult Schwann cells M. leprae appear to propagate the bacterial niche and maintain survival within the PNS. These studies may also provide new insights into our understanding of signaling mechanisms involve in both neurodegeneration and neuroregeneration. PMID- 20005918 TI - Toward a system-level understanding of microRNA pathway via mathematical modeling. AB - The microRNA (miRNA) pathway plays multiple roles in regulating mechanisms controlling both physiological and pathological processes such as the cell proliferation and cancers. But little is known about the dynamic properties, key rate-limiting steps as well as the stochastic noise in this pathway. Presently, a system-theoretic approach was presented to analyze and quantitative modeling of a generic miRNA pathway, which can be implemented deterministically and stochastically. Our results show that the inferred dynamic properties obtained from the mathematical models of the miRNA pathway are well consistent with previous experimental observations. By sensitivity analysis, the key steps in this pathway were found to be the miRNA gene transcription, RISC decay and mRNA formation. In addition, the results of quantified noise strength along the pathway demonstrate that the pathway can reduce the ingress noise and reveal the noise robustness property. Our findings also present testable hypothesis for experimental biologists to further investigate miRNA's increasing functional roles in regulating various cellular processes. PMID- 20005917 TI - Computational modeling of epithelial-mesenchymal transformations. AB - An epithelial-mesenchymal transformation (EMT) involves alterations in cell-cell and cell-matrix adhesion, the detachment of epithelial cells from their neighbors, the degradation of the basal lamina and acquisition of mesenchymal phenotype. Here we present Monte Carlo simulations for a specific EMT in early heart development: the formation of cardiac cushions. Cell rearrangements are described in accordance with Steinberg's differential adhesion hypothesis, which states that cells possess a type-dependent adhesion apparatus and are sufficiently motile to give rise to the tissue conformation with the largest number of strong bonds. We also implement epithelial and mesenchymal cell proliferation, cell type change and extracellular matrix production by mesenchymal cells. Our results show that an EMT is promoted more efficiently by an increase in cell-substrate adhesion than by a decrease in cell-cell adhesion. In addition to cushion tissue formation, the model also accounts for the phenomena of matrix invasion and mesenchymal condensation. We conclude that in order to maintain epithelial integrity during EMT the number of epithelial cells must increase at a controlled rate. Our model predictions are in qualitative agreement with available experimental data. PMID- 20005920 TI - Affect-related personality traits and contextual interference processing during perception of facial affect. AB - We investigated the influence of different personality dimensions on neural emotional conflict processing. A cluster analysis separated two different subsamples of subjects. One sample scored high on neuroticism, anxiety, depressivity and low on extraversion and was found to be highly sensitive to emotional conflicts during processing of negative and positive expressions. This sample revealed increased conflict induced activation in the amygdala during processing of negative expressions which was associated with higher levels of anxiety and depressivity. This amygdala activity was positively coupled with activity in dACC during processing of neutral expressions and negatively coupled in the sample with opposite personality scores. Depressivity was also associated with activation in subgenual ACC. Increased activation in subgenual ACC was also found during processing of positive expressions. This activation as well as activation in rostral ACC was higher in lower extraversion scores. In summary, individuals with higher neuroticism, anxiety, depressivity scores and low extraversion scores are sensitive to emotional conflicts during the processing of emotional expressions. Furthermore, emotional conflict resolution might challenge emotional regulative mechanisms in these subjects which became evident by increased activations in different subregions of the ACC. PMID- 20005919 TI - Gene networks and the neuroendocrine regulation of puberty. AB - A sustained increase in pulsatile release of gonadotrophin releasing hormone (GnRH) from the hypothalamus is an essential, final event that defines the initiation of mammalian puberty. This increase depends on coordinated changes in transsynaptic and glial-neuronal communication, consisting of activating neuronal and glial excitatory inputs to the GnRH neuronal network and the loss of transsynaptic inhibitory tone. It is now clear that the prevalent excitatory systems stimulating GnRH secretion involve a neuronal component consisting of excitatory amino acids (glutamate) and at least one peptide (kisspeptin), and a glial component that uses growth factors and small molecules for cell-cell signaling. GABAergic and opiatergic neurons provide transsynaptic inhibitory control to the system, but GABA neurons also exert direct excitatory effects on GnRH neurons. The molecular mechanisms that provide encompassing coordination to this cellular network are not known, but they appear to involve a host of functionally related genes hierarchically arranged. We envision that, as observed in other gene networks, the highest level of control in this network is provided by transcriptional regulators that, by directing expression of key subordinate genes, impose an integrative level of coordination to the neuronal and glial subsets involved in initiating the pubertal process. The use of high-throughput and gene manipulation approaches coupled to systems biology strategies should provide not only the experimental bases supporting this concept, but also unveil the existence of crucial components of network control not yet identified. PMID- 20005921 TI - CALHM1 P86L polymorphism does not alter amyloid-beta or tau in cerebrospinal fluid. AB - Recently, the P86L alteration in CALHM1 (calcium homeostasis modulator-1) was reported to be associated with Alzheimer's disease (AD). Moreover, the risk allele increased amyloid-beta (A beta) levels in conditioned media from cultured cells. Therefore, we hypothesized that CALHM1 P86L may modulate A beta or tau levels in cerebrospinal fluid (CSF). Nearly 200 individuals with AD or other cognitive disorders were included for CSF analysis and CALHM1 genotyping. No significant differences in CSF levels of A beta 42, tau or phospho-tau were found across the various CALHM1 genotypes. In conclusion, we found no evidence that CALHM1 P86L is associated with altered CSF levels of the investigated AD biomarkers. PMID- 20005922 TI - Caloric restriction provided after global ischemia does not reduce hippocampal cornu ammonis injury or improve functional recovery. AB - Since caloric restriction (CR) can modify multiple pathways central to the ischemic cascade and enhance neuroplasticity mechanisms, we hypothesized that CR should exert protective effects following brain ischemia. Previous studies have suggested benefit when CR was administered prior to ischemia. We investigated whether prolonged CR beginning after global ischemia would result in lasting protection as assessed by performance in the open field, as a measure of functional outcome, and hippocampal CA1 neuronal counts. Adult, male Mongolian gerbils were subjected to 5 min bilateral carotid artery occlusion (ISCH) or sham surgery (SHAM) with tympanic temperature maintained at 36.5+/-0.2 degrees C during the intra-ischemic period. After screening out gerbils with incomplete ischemia, each of the two surgical groups were randomly assigned to control diet (CON) or 30% CR for the duration of the study (60 d). Gerbils were tested in the open field on d3, 7, 10, 30 and 60. ISCH-CON animals showed a significantly higher level of activity in the open field (impaired habituation) compared to SHAM-CON gerbils on all test days (P<0.001). Open field activity was significantly lower in the ISCH-CR group than in ISCH-CON gerbils only on d7 (P=0.024). Open field activity of the SHAM-CR gerbils showed a trend to increase relative to that of SHAM-CON gerbils during the last 30 d of the study (P=0.055 on d60), raising the question of suitability of the open field test for long-term studies of CR and ischemia. Brain sections obtained at d60 were stained with hematoxylin and eosin. Hippocampal CA1 neuron counts were significantly reduced by ischemia (P<0.001), and there was no sparing effect of CR. Our findings suggest that prolonged 30% CR administered beginning after global ischemia cannot diminish brain injury or enhance long-term recovery. PMID- 20005924 TI - Modulation of AMPA excitatory postsynaptic currents in the spinal cord dorsal horn neurons by insulin. AB - Glutamate AMPA receptors are critical for sensory transmission at the spinal cord dorsal horn (DH). Plasma membrane AMPA receptor endocytosis that can be induced by insulin may underlie long term modulation of synaptic transmission. Insulin receptors (IRs) are known to be expressed on spinal cord DH neurons, but their possible role in sensory transmission has not been studied. In this work the effect of insulin application on fast excitatory postsynaptic currents (EPSCs) mediated by AMPA receptors evoked in DH neurons was evaluated. Acute spinal cord slices from 6 to 10 day old mice were used to record EPSCs evoked in visually identified superficial DH neurons by dorsal root primary afferent stimulation. AMPA EPSCs could be evoked in all of the tested neurons. In 75% of the neurons the size of the AMPA EPSCs was reduced to 62.1% and to 68.9% of the control values when 0.5 or 10 microM insulin was applied. There was no significant change in the size of the AMPA EPSCs in the remaining 25% of DH neurons. The membrane permeable protein tyrosine kinase inhibitor, lavendustin A (10 microM), prevented the insulin induced AMPA EPSC depression. Our results suggest a possible role of the insulin pathway in modulation of sensory and nociceptive synaptic transmission in the spinal cord. PMID- 20005923 TI - Projections from auditory cortex to midbrain cholinergic neurons that project to the inferior colliculus. AB - We have shown that auditory cortex projects to cholinergic cells in the pedunculopontine tegmental nucleus (PPT) and laterodorsal tegmental nucleus (LDT). PPT and LDT are the sources of cholinergic projections to the inferior colliculus, but it is not known if the cortical inputs contact the cholinergic cells that project to the inferior colliculus. We injected FluoroRuby into auditory cortex in pigmented guinea pigs to label cortical projections to PPT and LDT. In the same animals, we injected Fast Blue into the left or right inferior colliculus to label PPT and LDT cells that project to the inferior colliculus. We processed the brain to identify cholinergic cells with an antibody to choline acetyltransferase, which was visualized with a green fluorescent marker distinguishable from both FluoroRuby and Fast Blue. We then examined the PPT and LDT to determine whether boutons of FluoroRuby-labeled cortical axons were in close contact with cells that were double-labeled with the retrograde tracer and the immunolabel. Apparent contacts were observed ipsilateral and, less often, contralateral to the injected cortex. On both sides, the contacts were more numerous in PPT than in LDT. The results indicate that auditory cortex projects directly to brainstem cholinergic cells that innervate the ipsilateral or contralateral inferior colliculus. This suggests that cortical projections could elicit cholinergic effects on both sides of the auditory midbrain. PMID- 20005925 TI - Potential use of melatonergic drugs in analgesia: mechanisms of action. AB - Melatonin is a remarkable molecule with diverse physiological functions. Some of its effects are mediated by receptors while other, like cytoprotection, seem to depend on direct and indirect scavenging of free radicals not involving receptors. Among melatonin's many effects, its antinociceptive actions have attracted attention. When given orally, intraperitoneally, locally, intrathecally or through intracerebroventricular routes, melatonin exerts antinociceptive and antiallodynic actions in a variety of animal models. These effects have been demonstrated in animal models of acute pain like the tail-flick test, formalin test or endotoxin-induced hyperalgesia as well as in models of neuropathic pain like nerve ligation. Glutamate, gamma-aminobutyric acid, and particularly, opioid neurotransmission have been demonstrated to be involved in melatonin's analgesia. Results using melatonin receptor antagonists support the participation of melatonin receptors in melatonin's analgesia. However, discrepancies between the affinity of the receptors and the very high doses of melatonin needed to cause effects in vivo raise doubts about the uniqueness of that physiopathological interpretation. Indeed, melatonin could play a role in pain through several alternative mechanisms including free radicals scavenging or nitric oxide synthase inhibition. The use of melatonin analogs like the MT(1)/MT(2) agonist ramelteon, which lacks free radical scavenging activity, could be useful to unravel the mechanism of action of melatonin in analgesia. Melatonin has a promising role as an analgesic drug that could be used for alleviating pain associated with cancer, headache or surgical procedures. PMID- 20005926 TI - hGH-V gene expression and promoter activity under glucose and 5-azacytidine (5azaC) effects. AB - The metabolic conditions affecting placental development depend on nutritional state, genetic constitution and other external factors. The secretion of human placental growth hormone (hGH-V) had shown to be dependent of glucose, but the regulatory effects of this metabolite on hGH-V promoter activity and gene expression in presence of 5-azacytidine had not been studied. In this work we compared the hGH-V promoter activity and the endogenous mRNA expression in human placental choriocarcinoma cell line JAR in the presence of glucose and demethylating genome conditions. High glucose concentration in culture medium diminished hGH-V mRNA endogenous levels in JAR cells whereas the expression of hGH-V from transfected PACs was slightly higher; but in the presence of 5azaC a higher hGH-V gene expression from both the endogenous and the transfected ones was obtained. A drastic diminution of promoter analysis was shown when cells had no glucose (J0 cells) or in presence of 5azaC; the placental transcription factors that showed modified binding capacity were HES-2, PPAR-gamma, H4TF-1 and OCT-1. Our results suggest that in vitro suppressive glucose effect dictates a metabolic context to hGH-V gene expression and promoter regulation whereas a genomic methylation-dependent background is necessary to maintain placental transcription factors able to bind and regulate proximal promoter region of hGH-V in placental cells. PMID- 20005927 TI - Influence of amorphous content on compaction behaviour of anhydrous alpha lactose. AB - Modified lactoses are widely used as filler-binders in direct compression of tablets. Until today, little about the compaction behaviour of anhydrous alpha lactose is known. In this study, a new method to prepare anhydrous alpha-lactose from alpha-lactose monohydrate by desiccation with heated ethanol was evaluated and the influence of amorphous content in the lactose powder prior to modification on powder properties, compaction behaviour and storage stability was determined. The modification process led to anhydrous alpha-lactose with decreased bulk and tapped density, increased flow rate and significantly higher specific surface area. Due to the higher specific surface area, the compaction behaviour of the anhydrous alpha-lactose was found to be significantly better than the compaction behaviour of powder blends consisting of alpha-lactose monohydrate and amorphous lactose. An influence of the amorphous content prior to modification could be observed only at higher compaction forces. In general, tablets of modified powders needed longer time to disintegrate directly after compression. However, the storage stability of modified tablets was found to be better compared to the amorphous-crystalline tablets which were influenced by storage conditions, initial crushing strength as well as amorphous content due to the re-crystallization of amorphous lactose during storage. PMID- 20005928 TI - In vitro and in vivo evaluation of praziquantel loaded implants based on PEG/PCL blends. AB - In the present study, a series of praziquantel (PZQ) loaded implants based on PEG/PCL blends are fabricated by a combination of twin-screw mixing and hot-melt extrusion. In vitro drug release from these implants and the performance of the implants after implantation in rats are evaluated. XRD and DSC analysis results exhibit each component in the implants is mainly in its crystalline state. Dissolution test shows that the higher PEG content there is in the implants, the faster the drug will be released. Interestingly, PEG release from all implants is far faster than PZQ release, and complete PEG release occurs in 72 h. SEM result displays that after the in vitro drug release test, the cross-sections of implants with low PEG contents (0-5%) primarily consist of discrete pores; while those of implants with high PEG contents (10-30%) consist of interconnected pores or channels. The fitting results of drug release data with kinetic models reveal that PZQ release is governed by diffusion. After implantation, drug release becomes more moderate compared with in vitro drug release, and it tends to follow zero-order in the later stage. These results suggest that changing the composition of the PEG/PCL blends is an effective tool to adjust in vitro/in vivo drug release from the implants. PMID- 20005929 TI - Thin-coating as an alternative approach to improve flow properties of ibuprofen powder. AB - In the present study, thin-coating as a potential method for improving flow properties of cohesive ibuprofen powder was introduced. Briefly, the technique was based on the successive deposition of ultrasound-assisted fine polymer mist onto the surface of the powdered active pharmaceutical ingredient (API), producing individual particles with a hydrophilic thin-coat. A 0.15% m/V aqueous solution of hydroxypropyl methylcellulose (HPMC) was used. Particle size and surface analysis revealed a decrease in the cohesiveness of ibuprofen powder and an increase in the homogeneity of particle surfaces as a result of polymer treatment. Superficial changes caused a substantial improvement on the flowing characteristics of coated substance over uncoated. The enhancement in flow rate proceeded as the uniformity of the HPMC layer increased. In conclusion, the proposed technique is a simple and effective method that can be used as a continuous process to modify API particle surface properties, which in turn improve the handling of poorly flowable powder. PMID- 20005930 TI - Mixed micelle formation with phosphatidylcholines: the influence of surfactants with different molecule structures. AB - The number of mixed micellar (MM) drug products being introduced into the commercial pharmaceutical market is very limited although there is need for alternative dosage forms for poorly soluble active drug substances. While known systems are composed of phosphatidylcholine and bile salts, it was the aim of this study to investigate if alternative surfactants are able to form isotropically clear solutions over a broad range of concentrations and at higher ratios of phosphatidylcholine (PC). It was a particular challenge of this work to find a MM system with a unimodal particle size distribution since it is known that surfactants often form vesicles with phospholipids instead of MM. The theoretical approach behind this work was the transfer of the packing parameter concept, which describes the molecular association of one amphiphilic species, to the organisation behaviour of two different amphiphilic species (water-insoluble phospholipid+surfactant leading to MM). Therefore the influence of the surfactant molecular geometry on the ability to form MM with phospholipids was investigated. A homologous series of two different surfactant classes, namely polyglycerol esters and sucrose esters, with a large hydrophilic head region leading to a smaller packing parameter were analysed regarding their ability to form clear MM solutions with PC. For comparison, surfactants with no strictly defined partition between a polar head and a non-polar tail (e.g. Poloxamer 188) were tested. Decaglycerol laurate and especially sucrose laurate (SL) were superior compared to all other tested surfactants with respect to their ability to form clear solutions with hydrogenated PC (hPC) at a higher ratio and over a broad range of concentrations while unsaturated PC showed an inferior performance to form MM. The favourite MM system composed of SL with 0.5 weight fractions of hPC formed about 20 nm sized MM in a concentration range of 1.0-80 mg/mL and showing a unimodal particle size distribution with a PDI value <0.1. The results of the study have shown that the transferred packing parameter concept is applicable to the tested surfactants to describe their ability forming mixed micelles with PC. PMID- 20005931 TI - Recombinant human granulocyte colony stimulating factor pre-screening and screening of stabilizing carbohydrates and polyols. AB - Protein stabilization by solvent additives is frequently used concept in formulation development, although new technologies implemented over the past decade can improve protein biophysical as well as clinical properties by protein structural design (e.g. PEGylation, acylation, hesylation). The scope of this work was to evaluate the effect of chosen carbohydrate or polyol stabilizer in the formulation; firstly on linear peptide sequences on instable model of rHuG CSF cleaved macromolecule by novel method named protein and peptide stabilizer pre-screening PPSP (formulated tryptic digest mixture stability evaluation in 54 h) and on overall stability of rHuG-CSF macromolecule by quantifying all relevant degradation parameters. Comprehensive protein stabilizing screening study included conformational analysis of formulated rHuG-CSF protein to obtain information on its secondary structure conformational stability. Protein aggregation induced by modulating conditions in solution (e.g. thermal stress and agitation) was monitored over discrete time periods. Oxidation and deamidation, as well as truncation or hydrolysis were accurately quantified. Together with pre screening data, obtained by fast and resourceful amino acid sequence degradation analysis by LC-MS, statistical data evaluation of stabilizing contribution of substances selected from group of carbohydrates and polyols was performed. According to the statistical interpretation of obtained results the stabilizers were ranked in the following order: turanose, D-trehalose, lactitol, acetate buffer (non-stabilized sample), xylitol, cellobiitol, sorbitol, D-lyxose, leucrose, sorbitol without polysorbate, cellobiose. PMID- 20005932 TI - The freeze-thawed and freeze-dried stability of cytarabine-encapsulated multivesicular liposomes. AB - To investigate the stability of cytarabine-encapsulated multivesicular liposomes (MVLs) following freeze-thawing/freeze-drying, three types of phospholipids (EPC, DPPC, and DOPC) were separately employed to prepare MVLs using a double emulsification method. The cytarabine retention (CR), phase transition behavior, aggregation/rupture of vesicles and particle size were monitored using HPLC, differential scanning calorimetry (DSC), digital biological microscopy and a laser diffraction particle size analyzer. The effect of trehalose, the lipid bilayer composition and triglyceride on the drug retention was also investigated. The DPPC-MVLs and EPC-MVLs achieved the best protective effect during freeze thawing and freeze-drying, respectively, while DOPC-MVLs produced the lowest drug retention during both procedures. Digital biological microscopy showed that most of the MVLs were divided into small irregular and regular vesicles after freeze thawing and freeze-drying, which was in agreement with the reduction in particle size. The vesicle fragmentations may result from the splitting of triglyceride from the lipid membrane or rupture of the lipid membrane. The rehydrated EPC-MVLs still displayed a controlled-release profile in vitro, and the results presented in this work should help in stabilizing hydrophilic drug-encapsulated liposomes with a large particle size. PMID- 20005933 TI - Synthesis and characterization of functionalized poly(gamma-benzyl-L-glutamate) derivates and corresponding nanoparticles preparation and characterization. AB - For being fully efficient a targeted delivery system should associate simultaneously multiple functionalities. In this context, the association of several polymeric materials to form composite multifunctional particles can be foreseen. The present work describes the synthesis of different derivates of poly(gamma-benzyl-L-glutamate) and their use for the preparation of nanoparticles exhibiting different properties, including surface hydrophilization by PEG, fluorescence imaging by FITC and target recognition through easy attachment of desired ligands by using the avidin-biotin interaction, after the nanoparticles preparation. Four PBLG derivates were successfully obtained by ring-opening polymerization (ROP) of NCA, using various initiators corresponding to the molecules to be introduced into the copolymers. Further, nanoparticles smaller than 100 nm could be prepared using a nanoprecipitation technique and the presence of the active moieties introduced within the particles as well as their functionality has been checked. Very interestingly, it has been shown that biotin molecules could be efficiently introduced at the surface of the nanoparticles, which (for 75% of the theoretical amount) could be engaged in a complexation with avidin. It is suggested that this strategy offers the possibility to easily decorate these nanoparticles with various recognition ligands for specific targeting applications by using the well known biotin-avidin sandwich technique. PMID- 20005934 TI - Ascorbyl dipalmitate/PEG-lipid nanoparticles as a novel carrier for hydrophobic drugs. AB - L-ascorbyl 2,6-dipalmitate (ASC-DP), a fatty ester derivative of ascorbic acid, is poorly soluble in water and does not spontaneously form micelles or liposomal structures in water. In this study, we attempted to prepare an ASC-DP/surfactant nano-sized complex as a carrier for hydrophobic drugs. Samples were prepared by hydrating a solvent-evaporated film of ASC-DP/surfactant at a molar ratio of 1:1. Among the surfactants tested, distearoylphosphatidylethanolamine-polyethylene glycol 2000 (DSPE-PEG) was found to form stable nanoparticles with ASC-DP (average particle size: ca. 67 nm). Several hydrophobic drugs were incorporated in the ASC-DP/DSPE-PEG nanoparticles. Stability, toxicity, and blood residence of the drug-containing ASC-DP/DSPE-PEG nanoparticles were evaluated using amphotericin B (AmB) as the model drug. By intravenously administering mice with the formulations, we determined the minimum lethal dose of Fungizone, a formulation of AmB solubilized with sodium deoxycholate, was 3.0 mg/kg, while that of AmB/ASC-DP/DSPE-PEG nanoparticles was 10.0 mg/kg. When 2.0 mg/kg, Fungizone was administered, the mice showed higher renal and hepatic toxicities. Intravenously administered AmB/ASC-DP/DSPE-PEG nanoparticles demonstrated higher concentration in plasma than Fungizone. Thus, the ASC-DP/DSPE-PEG nanoparticle system appears to be a promising delivery system for hydrophobic drugs. PMID- 20005935 TI - The suitability of tris(hydroxylmethyl) aminomethane (THAM) as a buffering system for hydroxypropyl methylcellulose (HPMC) hydrophilic matrices containing a weak acid drug. AB - There are few studies of alkalising pH-modifiers in HPMC hydrophilic matrices. These agents may be incorporated to provide microenvironmental buffering and facilitate pH-independent release of weak acid drugs. This study compared tris(hydroxylmethyl) aminomethane (THAM, TRIS, tromethamine, trometamol) with sodium citrate as internal buffering agents for HPMC (4000 cps) 2208 and 2910 matrices containing felbinac, a weak acid drug which exhibits pH-dependent solubility. Drug release at pH 1.2 and 7.5 was accelerated by both buffers, but THAM-buffered matrices provided extended, diffusion-based release kinetics, without loss of matrix integrity at high buffer concentrations. Release kinetics appeared to be independent of media pH. THAM did not depress the sol-gel transition temperature or suppress HPMC particle swelling, and had minimal effects on gel layer formation. Sodium citrate promoted greater thickness of the early gel layer than THAM. Measurements of internal gel layer pH showed that both buffers produced a rapid alkalisation of the gel layer which was progressively lost. As result of its higher pK(a) and molar ratio on a percent weight basis, THAM provided a higher internal pH and a greater longevity of pH modification. It is concluded that THAM offers a useful buffering option for weak acid drugs in HPMC-based systems. PMID- 20005936 TI - PEG-PLA-PEG block copolymeric nanoparticles for oral immunization against hepatitis B. AB - PLA/PLGA nanoparticles are well known as efficient vaccine delivery systems, but they have got limitation in oral vaccine delivery because of their sensitivity to harsh gastric environment. The aim of present study was to improve the stability of PLA nanoparticles in such environment by copolymerizing PLA with PEG. Nanoparticles were formulated using different block copolymers AB, ABA and BAB (where 'A' is PLA and 'B' is PEG) encapsulating hepatitis B surface antigen (HBsAg) to evaluate their efficacy as oral vaccine delivery system. The results of in vitro studies engrave the efficiency of copolymeric nanoparticles to retain encapsulated antigen and average particle size even after 2 h incubation in simulated gastric fluid and simulated intestinal fluid. Fluorescence microscopic studies indicated efficient uptake of copolymeric nanoparticles by gut mucosa of immunized mice model as compared to control. Finally copolymeric and PLA nanoparticles, encapsulating HBsAg, were evaluated for their adjuvancity in generating immune response after oral administration. PLA nanoparticles could not generate an effective immune response due to stability issues. On the other hand, oral administration of copolymeric nanoparticles exhibited effective levels of humoral immunity along with the mucosal (sIgA) and cellular immune response (T(H)1). The results of in vitro and in vivo studies demonstrate that BAB nanoparticles depict enhanced mucosal uptake leading to effective immune response as compared to other copolymeric nanoparticles. Present study indicates the efficacy of BAB nanoparticles as a promising carrier for oral immunization. PMID- 20005937 TI - Process optimization, characterization and evaluation in vivo of oxymatrine phospholipid complex. AB - The objective of this study was to prepare oxymatrine-phospholipid complex (OMT PLC) to enhance oral bioavailability of oxymatrine. A central composite design approach was used for process optimization. The physicochemical properties of the complex obtained by optimal parameters were investigated by means of differential scanning calorimetry (DSC), X-ray diffraction (XRD) and N-octanol/water partition coefficient. Compared with those of the physical mixture or oxymatrine, the hepatocytes permeability of oxymatrine-phospholipid complexes was studied. The concentrations of oxymatrine after oral administration of OMT-PLC at different time in rats were determined by HPCE. Multiple linear regression analysis for process optimization revealed that the acceptable OMT-PLC was obtained wherein the optimal values of X(1), X(2) and X(3) were 3, 60 degrees C and 3 h, respectively. The oxymatrine and phospholipids in the OMT-PLC were combined by non-covalent bond, not forming a new compound. The better hepatocytes permeability was obtained by the OMT-PLC. Pharmacokinetic parameters of the complex in rats were T(max) 2.17 h, C(max) 0.437 microg ml(-1), AUC(0-infinity) 9.43 microg h ml(-1), respectively. The bioavailability of oxymatrine in rats was increased remarkably after oral administration of OMT-PLC (p<0.05), compared with those of oxymatrine or the physical mixture. This was mainly due to an improvement of the solubility of OMT-PLC. PMID- 20005938 TI - Masking of rippled-spectrum-pattern resolution in diotic and dichotic presentations. AB - Using rippled noise probes, spectrum-pattern resolution was measured with and without a narrow-band noise masker. Diotic presentation of both the probe and masker (S(0)N(0) mode) resulted in decreased spectrum resolution as compared to the control (no masker) conditions. The effects of the low- and on-frequency maskers differed quantitatively, however in both cases the ability to discriminate the probe spectrum pattern was suppressed completely when the masker/probe level ratio exceeded 10dB (on-frequency masker) or 10-25dB, depending on the probe level (low-frequency masker). The effect of the high frequency masker was negligible. Slight but noticeable releasing of the spectrum pattern resolution was found when the probe was presented to both ears in-phase and the masker counter-phase (S(0)N(pi) mode). In conditions of the probe delivered to one ear and the masker to the other ear (S(L)N(R) mode), the effect on the spectrum-pattern resolution was slight or negligible within a wide range of the noise/probe ratio. PMID- 20005939 TI - The floating mass transducer at the round window: direct transmission or bone conduction? AB - The round window placement of a floating mass transducer (FMT) is a new approach for coupling an implantable hearing system to the cochlea. We evaluated the vibration transfer to the cochlear fluids of an FMT placed at the round window (rwFMT) with special attention to the role of bone conduction. A posterior tympanotomy was performed on eleven ears of seven human whole head specimens. Several rwFMT setups were examined using laser Doppler vibrometry measurements at the stapes and the promontory. In three ears, the vibrations of a bone anchored hearing aid (BAHA) and an FMT fixed to the promontory (pFMT) were compared to explore the role of bone conduction. Vibration transmission to the measuring point at the stapes was best when the rwFMT was perpendicularly placed in the round window and underlayed with connective tissue. Fixation of the rwFMT to the round window exhibited significantly lower vibration transmission. Although measurable, bone conduction from the pFMT was much lower than that of the BAHA. Our results suggest that the rwFMT does not act as a small bone anchored hearing aid, but instead, acts as a direct vibratory stimulator of the round window membrane. PMID- 20005940 TI - Activation of human neutrophils by titanium dioxide (TiO2) nanoparticles. AB - This paper describes the in vitro effects of titanium dioxide (TiO(2)) nanoparticles (NPs) upon human neutrophils. Kinetic experiments revealed no cell necrosis after 24h of treatment with TiO(2) (0-100 microg/ml). In contrast, TiO(2)-induced change in cellular morphology in a concentration-dependent manner in neutrophils over time, indicating its potential to activate these cells. To further support this, we demonstrated that TiO(2) markedly and rapidly induced tyrosine phosphorylation events, including phosphorylation of two key enzymes, p38 mitogen-activated protein kinase (MAPK) and extracellular signal-regulated kinases-1/2 (Erk-1/2). We also determined the effects of TiO(2) on two neutrophil functions requiring a longer exposure period between NPs and cells: apoptosis and cytokine production. Interestingly, at concentrations >or=20 microg/ml, TiO(2) inhibited neutrophil apoptosis in a concentration-dependent manner after 24h of treatment. Supernatants from TiO(2)-induced neutrophils were harvested after 24h and tested for the presence of 36 different analytes (cytokines, chemokines) using an antibody array assay. TiO(2) treatment increased production of 13 (36%) analytes, including IL-8, which exhibited the greatest increase ( approximately 16 x control cell levels). The increased production of IL-8 was confirmed by ELISA. We conclude that TiO(2) exerts important neutrophil agonistic properties in vitro. PMID- 20005941 TI - Cell-density-dependent methylmercury susceptibility of cultured human brain microvascular pericytes. AB - The knowledge of vascular toxicity is important for understanding the neurotoxicity of methylmercury. In the present study, we investigated the cell density-dependent susceptibility of human brain microvascular pericytes to methylmercury-induced toxicity by using a cell-culture system. The susceptibility of sparse pericyte cultures to methylmercury was greater than that of the dense cultures. In addition, the sparse cultures were more susceptible to methylmercury than to inorganic mercury and cadmium. The intracellular accumulation of methylmercury in the sparse cells was significantly higher than that in the dense cells. Methylmercury is transported through the L-type large neutral amino acid transporter (LAT 1) in the form of a complex with cysteine. The mRNA- and protein level expressions of LAT 1 in the sparse cells were markedly higher than those in the dense cells; in addition, the LAT 1 expression was increased by methylmercury. However, there was no reduction in the levels of glutathione and metallothionein, which are involved in the defense mechanisms against methylmercury, in the sparse cells. The present data revealed that pericytes are markedly susceptible to methylmercury-induced cytotoxicity at low cell densities. The susceptibility of the sparse pericytes is postulated to be due to the not only constitutively higher but also methylmercury-induced expression of LAT 1, which increased the intracellular accumulation of methylmercury. PMID- 20005942 TI - Oleanolic acid and ursolic acid induce apoptosis in four human liver cancer cell lines. AB - Apoptotic effects of oleanolic acid (OA) and ursolic acid (UA) on human liver cancer HepG2, Hep3B, Huh7 and HA22T cell lines were examined. OA or UA at 2, 4, 8 micromol/L were used and their effects on cell viability, DNA fragmentation, mitochondrial membrane potential (MMP), activity of Na(+)-K(+)-ATPase, caspase-3 and caspase-8, cell adhesion, level of intercellular adhesion molecule (ICAM)-1 and vascular endothelial growth factor (VEGF) in these cell lines were determined. OA or UA treatments concentration-dependently decreased cell viability and increased DNA fragmentation in HepG2 and Hep3B cell lines (P<0.05). However, these two compounds reduced viability and increased DNA fragmentation in Huh7 cell only at 4 and 8 micromol/L (P<0.05). OA or UA treatments concentration dependently lowered MMP in HepG2, Hep3B and HA22T cell lines (P<0.05). These two compounds also concentration-dependently diminished Na(+)-K(+)-ATPase activity and VEGF level in four test cell lines (P<0.05). Besides Huh7 cell, OA or UA treatments concentration-dependently elevated caspase-3 and caspase-8 activities in other three cell lines (P<0.05). Besides HA22T cell, these two compounds concentration-dependently inhibited cell adhesion and decreased ICAM-1 level in other three cell lines (P<0.05). These findings support that OA and UA are potent anti-cancer agents to cause apoptosis in these liver cancer cell lines. PMID- 20005943 TI - EBV transformation and cell culturing destabilizes DNA methylation in human lymphoblastoid cell lines. AB - Recent research suggests that epigenetic alterations involving DNA methylation can be causative for neurodevelopmental, growth and metabolic disorders. Although lymphoblastoid cell lines have been an invaluable resource for the study of both genetic and epigenetic disorders, the impact of EBV transformation, cell culturing and freezing on epigenetic patterns is unknown. We compared genome-wide DNA methylation patterns of four white blood cell samples, four low-passage lymphoblastoid cell lines pre and post freezing and four high-passage lymphobastoid cell lines, using two microarray platforms: Illumina HumanMethylation27 platform containing 27,578 CpG sites and Agilent Human CpG island Array containing 27,800 CpG islands. Comparison of genome-wide methylation profiles between white blood cells and lymphoblastoid cell lines demonstrated methylation alterations in lymphoblastoid cell lines occurring at random genomic locations. These changes were more profound in high-passage cells. Freezing at low-passages did not have a significant effect on DNA methylation. Methylation changes were observed in several imprinted differentially methylated regions, including DIRAS3, NNAT, H19, MEG3, NDN and MKRN3, but not in known imprinting centers. Our results suggest that lymphoblastoid cell lines should be used with caution for the identification of disease-associated DNA methylation changes or for discovery of new imprinted genes, as the methylation patterns seen in these cell lines may not always be representative of DNA methylation present in the original B-lymphocytes of the patient. PMID- 20005944 TI - Inhibition of SREBP-1c-mediated hepatic steatosis and oxidative stress by sauchinone, an AMPK-activating lignan in Saururus chinensis. AB - Sauchinone, as an AMP-activated kinase (AMPK)-activating lignan in Saururus chinensis, has been shown to prevent iron-induced oxidative stress and liver injury. Sterol regulatory element binding protein-1c (SREBP-1c) plays a key role in hepatic steatosis, which promotes oxidative stress in obese subjects. Previously, we identified the role of AMPK in liver X receptor-alpha (LXRalpha) mediated SREBP-1c-dependent lipogenesis. Because sauchinone as an antioxidant has the ability to activate AMPK, this study investigated its effects on SREBP-1c dependent lipogenesis in hepatocytes and in high-fat diet (HFD)-induced hepatic steatosis and oxidative injury. Sauchinone prevented the ability of an LXRalpha agonist (T0901317) to activate SREBP-1c, repressing transcription of the fatty acid synthase, acetyl-CoA carboxylase, stearoyl-CoA desaturase-1, ATP-binding cassette transporter A1, and LXRalpha genes. Consistent with this, an HFD in mice caused fat accumulation in the liver with SREBP-1c induction, which was attenuated by sauchinone treatment. Also, sauchinone had the ability to inhibit oxidative stress as shown by decreases in thiobarbituric acid-reactive substance formation, nitrotyrosinylation, and 4-hydroxynonenal production. Moreover, it prevented not only the liver injury, but also the AMPK inhibition elicited by HFD feeding. These results demonstrate that sauchinone has the capability to inhibit LXRalpha-mediated SREBP-1c induction and SREBP-1c-dependent hepatic steatosis, thereby protecting hepatocytes from oxidative stress induced by fat accumulation. PMID- 20005945 TI - Modulation of redox status and calcium handling by extremely low frequency electromagnetic fields in C2C12 muscle cells: A real-time, single-cell approach. AB - The biological effects of electric and magnetic fields, which are ubiquitous in modern society, remain poorly understood. Here, we applied a single-cell approach to study the effects of short-term exposure to extremely low frequency electromagnetic fields (ELF-EMFs) on muscle cell differentiation and function using C2C12 cells as an in vitro model of the skeletal muscle phenotype. Our focus was on markers of oxidative stress and calcium (Ca(2+)) handling, two interrelated cellular processes previously shown to be affected by such radiation in other cell models. Collectively, our data reveal that ELF-EMFs (1) induced reactive oxygen species production in myoblasts and myotubes with a concomitant decrease in mitochondrial membrane potential; (2) activated the cellular detoxification system, increasing catalase and glutathione peroxidase activities; and (3) altered intracellular Ca(2+)homeostasis, increasing the spontaneous activity of myotubes and enhancing cellular reactivity to a depolarizing agent (KCl) or an agonist (caffeine) of intracellular store Ca(2+)channels. In conclusion, our data support a possible link between exposure to ELF-EMFs and modification of the cellular redox state, which could, in turn, increase the level of intracellular Ca(2+)and thus modulate the metabolic activity of C2C12 cells. PMID- 20005946 TI - Extracellular superoxide dismutase polymorphism in mice: Allele-specific effects on phenotype. AB - Extracellular superoxide dismutase (ecSOD) protects the extracellular matrix from oxidative stress. We previously reported a new allele for ecSOD, expressed in 129P3/J mice (129), which differs from the wild type (wt), expressed in C57BL/6J and other strains, by two amino acid substitutions and a 10-bp deletion in the 3' UTR of the mRNA (A. Pierce et al., 2003, Arterioscler. Thromb. Vasc. Biol.23:1820 1825). The newly discovered allele is associated with a phenotype of significantly increased circulating and heparin-releasable enzyme activities and levels. To examine the properties of the two forms of ecSOD in an identical environment we generated, by extensive backcrossing of ecSOD heterozygous progeny to C57BL/6J females, a congenic C57 strain with the 129 (or wt) allele of ecSOD. These mice are homozygous for nearly 5000 SNPs across all chromosomes, as determined by the Affymetrix Parallele Mouse 5K SNP panel. This study describes the generation of the congenic mice (genetically >99.8% identical) and their ecSOD phenotype. The congenic mouse plasma ecSOD activity before and after heparin administration recapitulates the differences reported in the founder mice. Tissue enzyme distribution is similar in both congenic groups, although the 129 allele is associated with higher levels of enzyme expression despite lower levels of enzyme mRNA. In these characteristics the phenotype is allele driven, with little impact from the rest of the genome. The congenic mice carrying the 129 allele have mRNA levels that are in between those in the founder 129P3/J and C57BL/6J strains. We conclude that the ecSOD phenotype in most aspects of enzyme expression is allele driven, with the exception of tissue mRNA levels, for which a significant contribution by the surrounding (host) genome is observed. These results also suggest potential allele-specific differences in the regulation of ecSOD synthesis and intracellular processing/secretion of ecSOD, independent of the genotype context. Most importantly, the congenic mice offer an excellent model to examine the regulatory mechanisms of ecSOD expression and the role of ecSOD in various diseases involving oxidative stress. PMID- 20005947 TI - Surface dental enamel lead levels and antisocial behavior in Brazilian adolescents. AB - Lead poisoning has been reportedly linked to a high risk of learning disabilities, aggression and criminal offenses. To study the association between lead exposure and antisocial/delinquent behavior, a cross-sectional study was conducted with 173 Brazilian youths aged 14-18 and their parents (n=93), living in impoverished neighborhoods of Bauru-SP, with high criminality indices. Self Reported Delinquency (SRD) and Child Behavior Checklist (CBCL) questionnaires were used to evaluate delinquent/antisocial behavior. Body lead burdens were evaluated in surface dental enamel acid microbiopsies. The dental enamel lead levels (DELL) were quantified by graphite furnace atomic absorption spectrometry (GFAAS) and phosphorus content was measured using inductively coupled plasma optical emission spectrometry (ICP-OES). Logistic regression was used to identify associations between DELL and each scale defined by CBCL and SRD scores. Odd ratios adjusted for familial and social covariates, considering a group of youths exposed to high lead levels (>or=75 percentile), indicated that high DELL is associated with increased risk of exceeding the clinical score for somatic complaints, social problems, rule-breaking behavior and externalizing problems (CI 95%). High DELL was not found to be associated with elevated SRD scores. In conclusion, our data support the hypothesis that high-level lead exposure can trigger antisocial behavior, which calls for public policies to prevent lead poisoning. PMID- 20005948 TI - A two-layer diffusive model for describing the variability of transdermal drug permeation. AB - There is mounting evidence that the permeability coefficients (k(p)) that describe any given transdermal drug permeation process generally follow some form of positively skewed, non-symmetrical distribution rather than a simple normal distribution. Yet a suitable theoretical treatment of this area has not been undertaken to date. In this paper, we describe a two-layer model that can explain five drugs'k(p) variabilities as measured in two previously published papers. The model shows why rapidly permeating drugs would tend to exhibit more symmetrical k(p) distributions while progressively more slowly permeating drugs would tend to exhibit progressively more positively skewed k(p) distributions. Future research should take this effect into account when comparing the flux variabilities of hydrophilic and lipophilic drugs. PMID- 20005949 TI - Gene identification and proteomic analysis of the esterases of the cotton bollworm, Helicoverpa armigera. AB - Some of the resistance of Helicoverpa armigera to conventional insecticides such as organophosphates and synthetic pyrethroids appears to be due to metabolic detoxification by carboxylesterases. To investigate the H. armigera carboxyl/cholinesterases, we created a data set of 39 putative paralogous H. armigera carboxyl/cholinesterase sequences from cDNA libraries and other sources. Phylogenetic analysis revealed a close relationship between these sequences and 70 carboxyl/cholinesterases from the recently sequenced genome of the silkworm, Bombyx mori, including several conserved clades of non-catalytic proteins. A juvenile hormone esterase candidate from H. armigera was identified, and B. mori orthologues were proposed for 31% of the sequences examined, however low similarity was found between lepidopteran sequences and esterases previously associated with insecticide resistance from other insect orders. A proteomic analysis of larval esterases then enabled us to match seven of the H. armigera carboxyl/cholinesterase sequences to specific esterase isozymes. All identified sequences were predicted to encode catalytically active carboxylesterases, including six proteins with N-terminal signal peptides and N-glycans, with two also containing C-terminal signals for glycosylphosphatidylinositol anchor attachment. Five of these sequences were matched to zones of activity on native PAGE at relative mobility values previously associated with insecticide resistance in this species. PMID- 20005950 TI - Native subunit composition of two insect nicotinic receptor subtypes with differing affinities for the insecticide imidacloprid. AB - Neonicotinoid insecticides, such as imidacloprid, are selective agonists of insect nicotinic acetylcholine receptors (nAChRs) and are used extensively to control a variety of insect pest species. The brown planthopper (Nilaparvata lugens), an insect pest of rice crops throughout Asia, is an important target species for control with neonicotinoid insecticides such as imidacloprid. Studies with nAChRs purified from N. lugens have identified two [(3)H]imidacloprid binding sites with different affinities (K(d) = 3.5 +/- 0.6 pM and 1.5 +/- 0.2 nM). Co-immunoprecipitation studies with native preparations of N. lugens nAChRs, using subunit-selective antisera, have demonstrated the co-assembly of Nlalpha1, Nlalpha2 and Nlbeta1 subunits into one receptor complex and of Nlalpha3, Nlalpha8 and Nlbeta1 into another. Immunodepletion of Nlalpha1 or Nlalpha2 subunits resulted in the selective loss of the lower affinity imidacloprid binding site, whereas immunodepletion of Nlalpha3 or Nlalpha8 caused the selective loss of the high-affinity site. Immunodepletion of Nlbeta1 resulted in a complete absence of specific imidacloprid binding. In contrast, immunodepletion with antibodies selective for other N. lugens nAChR subunits (Nlalpha4, Nlalpha6, Nlalpha7 and Nlbeta2) had no significant effect on imidacloprid binding. Taken together, these data suggest that nAChRs containing Nlalpha1, Nlalpha2 and Nlbeta1 constitute the lower affinity binding site, whereas nAChRs containing Nlalpha3, Nlalpha8 and Nlbeta1 constitute the higher affinity binding site for imidacloprid in N. lugens. PMID- 20005951 TI - Inflammation in neuropsychiatric disease. PMID- 20005953 TI - Kindling as a model of temporal lobe epilepsy induces bilateral changes in spontaneous striatal activity. AB - Basal ganglia are engaged in seizure propagation, control of seizures, and in epilepsy-induced neuroplasticity. Here, we tested the hypothesis that previously observed histological and neurochemical changes in the striatum of amygdala kindled rats as a model of temporal lobe epilepsy are reflected in alterations of spontaneous striatal firing rates and patterns. Because experimental histological and clinical imaging studies indicated a bilateral involvement of the striatum in epilepsy-induced neuroplasticity, in vivo single-unit recordings were done bilaterally 1 day after a kindled seizure in rats kindled via the right amygdala. Compared to control animals, we observed (1) an increased irregularity of firing of neurons classified as striatal projection neurons and located in the anterior striatum ipsilateral to the kindling side and (2) an increased spontaneous activity of neurons classified as striatal projection neurons and located in the anterior striatum contralateral to the kindling side. These hyperactive neurons were located within the dorsolateral (sensorimotor) subregion of the striatum. The present study represents the first evidence of kindling-induced bilateral changes in electrophysiological properties of striatal neurons and demonstrates that the striatum is strongly affected by the functional reorganization of neurocircuits associated with kindling. The changes are probably caused by a combination of several factors including disturbed bilateral limbic and neocortical input as well as disturbed intrastriatal GABAergic function. The changes reflect a pathophysiological state predisposing the brain to epileptic discharge propagation or else (contralateral striatum) could represent a compensatory network of inhibitory circuits activated to prevent the propagation of seizure activity. The findings are relevant for a better understanding of kindling-induced network changes and might provide new targets for therapeutic manipulations in epilepsies. PMID- 20005955 TI - Dexamethasone protects human fibroblasts from apoptosis via an S1P3-receptor subtype dependent activation of PKB/Akt and Bcl XL. AB - Topical used glucocorticoids (GC) represent an important class of steroid hormones for the treatment of a broad range of acute or chronic inflammatory diseases. Most interestingly, GC exert a pronounced anti-apoptotic effect in primary human fibroblasts whereas in variety of hematopoietic cells a pro apoptotic effect is visible. Recently, it has been discovered that in human fibroblasts the GC dexamethasone (Dex) exerts its protection from programmed cell death via the formation of the lipid mediator sphingosine 1-phosphate (S1P) followed by an activation of the S1P(3)-receptor subtype. In the present study, the molecular mechanism of Dex to protect human fibroblasts from apoptosis was elucidated. Thereupon, Dex not only mediates its anti-apoptotic effect via activation of phosphoinositide 3-kinase (PI3K)/Akt signalling but also includes an involvement of the Bcl-2 family protein Bcl(XL). Most interestingly, the use of S1P(3)-knockout fibroblasts revealed that the S1P(3)-receptor subtype is crucial for activation of PKB/Akt as well as Bcl(XL) by Dex. PMID- 20005954 TI - Induced secretion of beta-hexosaminidase by human brain endothelial cells: a novel approach in Sandhoff disease? AB - Sandhoff disease is an autosomal recessive lysosomal disorder due to mutations in the beta-hexosaminidase beta-chain gene, resulting in beta-hexosaminidases A (alphabeta) and B (betabeta) deficiency and GM2 ganglioside accumulation in the brain. In this study, our aim was to demonstrate that transduction of cerebral endothelial cells cultured in two-chamber culture inserts with a lentiviral vector encoding the hexosaminidases alpha and beta chains could induce a vectorial secretion of hexosaminidases. Therefore, the human cerebral endothelial cell line hCMEC/D3 was infected with the bicistronic vector from the apical compartment, and beta-hexosaminidase activity was measured in transduced cells and in deficient fibroblasts co-cultured in the basal (i.e. brain) compartment. Induced beta-hexosaminidase secretion by transduced hCMEC/D3 cells was sufficient to allow for a 70-90% restoration of beta-hexosaminidase activity in deficient fibroblasts. On the basis of these in vitro data, we propose that brain endothelium be considered as a novel therapeutic target in Sandhoff disease. PMID- 20005956 TI - Involvement of angiotensin II in intestinal cholesterol absorption. AB - Niemann-Pick C1-like 1 (NPC1L1) protein is identified as a key molecule of cholesterol absorption into the intestine. Although there is a controversy about the association between sitosterol levels and cardiovascular disease (CVD), cholesterol absorption may contribute to the increased risk for CVD because increased levels of sitosterol, a marker of cholesterol absorption, are associated with future cardiovascular events in high-risk patients. However, which anthropometric and metabolic variables could regulate serum levels of sitosterol in humans and whether serum sitosterol levels might reflect transport function of NPC1L1 are largely unknown. In this study, we first investigated the independent determinants of serum sitosterol levels in apparently healthy patients not taking lipid-lowering agents. We next examined the effects of angiotensin II on NPC1L1 gene and protein expression in differentiated Caco-2 cells. Seventy apparently health patients not taking lipid-lowering agents (28 men and 42 women, mean age 73.7+/-10.1 years old) underwent a complete history and physical examination, determination of blood chemistries, including serum levels of sitosterol. Univariate regression analysis showed that serum levels of sitosterol were associated with low-density-lipoprotein (LDL)-cholesterol (r=0.284, p=0.021) and use of the renin-angiotensin system (RAS) inhibitors (r= 0.289, p=0.018). By the use of multiple stepwise regression analyses, use of RAS inhibitors (p=0.025) was remained significant independently. Further, angiotensin II was found to up-regulate NPC1L1 mRNA and protein levels in Caco-2 cells, which were completely blocked by an angiotensin II type 1 receptor blocker or an anti oxidant, N-acetylcysteine. The present study suggests the possible involvement of RAS in NPC1L1 expression in vitro and cholesterol absorption in humans. PMID- 20005957 TI - Modest proteasomal inhibition by aberrant ubiquitin exacerbates aggregate formation in a Huntington disease mouse model. AB - UBB(+1), a mutant form of ubiquitin, is both a substrate and an inhibitor of the proteasome which accumulates in the neuropathological hallmarks of Huntington disease (HD). In vitro, expression of UBB(+1) and mutant huntingtin synergistically increase aggregate formation and polyglutamine induced cell death. We generated a UBB(+1) transgenic mouse line expressing UBB(+1) within the neurons of the striatum. In these mice lentiviral driven expression of expanded huntingtin constructs in the striatum results in a significant increase in neuronal inclusion formation. Although UBB(+1) transgenic mice show neither a decreased lifespan nor apparent neuronal loss, they appear to be more vulnerable to toxic insults like expanded polyglutamine proteins due to a modest proteasome inhibition. These findings underscore the relevance of an efficient ubiquitin proteasome system in HD. PMID- 20005958 TI - Helical image reconstruction of the outward-open human erythrocyte band 3 membrane domain in tubular crystals. AB - The C-terminal membrane domain of erythrocyte band 3 functions as an anion exchanger. Here, we report the three-dimensional (3D) structure of the membrane domain in an inhibitor-stabilized, outward-open conformation at 18A resolution. Unstained, frozen-hydrated tubular crystals containing the membrane domain of band 3 purified from human red blood cells (hB3MD) were examined using cryo electron microscopy and iterative helical real-space reconstruction (IHRSR). The 3D image reconstruction of the tubular crystals showed the molecular packing of hB3MD dimers with dimensions of 60 x 110 A in the membrane plane and a thickness of 70A across the membrane. Immunoelectron microscopy and carboxyl-terminal digestion demonstrated that the intracellular surface of hB3MD was exposed on the outer surface of the tubular crystal. A 3D density map revealed that hB3MD consists of at least two subdomains and that the outward-open form is characterized by a large hollow area on the extracellular surface and continuous density on the intracellular surface. PMID- 20005959 TI - Structural and functional characterization of H+, K+-ATPase with bound fluorinated phosphate analogs. AB - Gastric H(+),K(+)-ATPase is responsible for gastric acid secretion. In order to characterize the phosphorylation events on H(+),K(+)-ATPase, the properties of fluorinated phosphate analogs [XFs, e.g. aluminum fluoride (AlF), beryllium fluoride (BeF) and magnesium fluoride (MgF)], and the structural differences induced by XFs were investigated. The addition of divalent cations to the XF inhibited H(+),K(+)-ATPase restores the activity of the AlF- or MgF-inhibited, but not of the BeF-inhibited enzyme, although limited trypsin digestion reveals that they assume the same E(2)P-like state. To clarify the conformational differences induced by XFs, the structure of BeF-bound H(+),K(+)-ATPase was analyzed at 8A resolution. The structure is almost identical to the previously reported AlF-bound E(2)P structure, unlike the distinctive X-ray structure of BeF bound SERCA, in which the luminal gate was observed to be widely opened. Since the analyzed structure of the H(+),K(+)-ATPase revealed that both AlF and BeF bound to the P domain were not exposed to the solvent, the dissociation of XFs induced by divalent cations could be interpreted in terms of stability against thermal fluctuations. Furthermore, the conformational differences found between the cytoplasmic domains of H(+),K(+)-ATPase and SERCA provide a framework to understand the characteristic mechanism, by which divalent cations reactivate the XF-inhibited H(+),K(+)-ATPase. PMID- 20005952 TI - Convergent evidence for abnormal striatal synaptic plasticity in dystonia. AB - Dystonia is a functionally disabling movement disorder characterized by abnormal movements and postures. Although substantial recent progress has been made in identifying genetic factors, the pathophysiology of the disease remains a mystery. A provocative suggestion gaining broader acceptance is that some aspect of neural plasticity may be abnormal. There is also evidence that, at least in some forms of dystonia, sensorimotor "use" may be a contributing factor. Most empirical evidence of abnormal plasticity in dystonia comes from measures of sensorimotor cortical organization and physiology. However, the basal ganglia also play a critical role in sensorimotor function. Furthermore, the basal ganglia are prominently implicated in traditional models of dystonia, are the primary targets of stereotactic neurosurgical interventions, and provide a neural substrate for sensorimotor learning influenced by neuromodulators. Our working hypothesis is that abnormal plasticity in the basal ganglia is a critical link between the etiology and pathophysiology of dystonia. In this review we set up the background for this hypothesis by integrating a large body of disparate indirect evidence that dystonia may involve abnormalities in synaptic plasticity in the striatum. After reviewing evidence implicating the striatum in dystonia, we focus on the influence of two neuromodulatory systems: dopamine and acetylcholine. For both of these neuromodulators, we first describe the evidence for abnormalities in dystonia and then the means by which it may influence striatal synaptic plasticity. Collectively, the evidence suggests that many different forms of dystonia may involve abnormal plasticity in the striatum. An improved understanding of these altered plastic processes would help inform our understanding of the pathophysiology of dystonia, and, given the role of the striatum in sensorimotor learning, provide a principled basis for designing therapies aimed at the dynamic processes linking etiology to pathophysiology of the disease. PMID- 20005960 TI - A different conformation for linker L12 in IF molecules in the molecular and filamentous forms: an hypothesis. AB - The rod domain of IF molecules has been characterized as four alpha-helical coiled-coil segments (1A, 1B, 2A and 2B), three linkers (L1, L12 and L2) and a stutter at the centre of segment 2B. Two of these breaks in coiled-coil continuity (L2 and stutter) have been modelled on the basis of structural data obtained from related proteins. Subsequently, X-ray crystallographic studies on fragments of IF molecules have shown that both models were correct. The third of the breaks - L1 - was predicted to have a flexible structure, consistent with observations that the head domain can fold back over segments 1A and 1B and also unwind into separate strands. Here the structure of the fourth discontinuity (L12) has been modelled. For most IF chain types two conformations are proposed for an eight-residue motif that displays a quasi two-residue repeat based on the presence of apolar residues. In IF it is proposed that the motif will adopt an alpha-helical conformation but that in the molecule the conformation will be beta like. Thus, assembly will result in or result from a conformational change in L12 thereby attributing L12 a more dynamic and important role in assembly than expected. PMID- 20005962 TI - Modulating cortical connectivity in stroke patients by rTMS assessed with fMRI and dynamic causal modeling. AB - Data derived from transcranial magnetic stimulation (TMS) studies suggest that transcallosal inhibition mechanisms between the primary motor cortex of both hemispheres may contribute to the reduced motor performance of stroke patients. We here investigated the potential of modulating pathological interactions between cortical motor areas by means of repetitive TMS using functional magnetic resonance imaging (fMRI) and dynamic causal modeling (DCM). Eleven subacute stroke patients were scanned 1-3 months after symptom onset while performing whole hand fist closure movements. After a baseline scan, patients were stimulated with inhibitory 1-Hz rTMS applied over two different locations: (i) vertex (control stimulation) and (ii) primary motor cortex (M1) of the unaffected (contralesional) hemisphere. Changes in the endogenous and task-dependent effective connectivity were assessed by DCM of a bilateral network comprising M1, lateral premotor cortex, and the supplementary motor area (SMA). The results showed that rTMS applied over contralesional M1 significantly improved the motor performance of the paretic hand. The connectivity analysis revealed that the behavioral improvements were significantly correlated with a reduction of the negative influences originating from contralesional M1 during paretic hand movements. Concurrently, endogenous coupling between ipsilesional SMA and M1 was significantly enhanced only after rTMS applied over contralesional M1. Therefore, rTMS applied over contralesional M1 may be used to transiently remodel the disturbed functional network architecture of the motor system. The connectivity analyses suggest that both a reduction of pathological transcallosal influences (originating from contralesional M1) and a restitution of ipsilesional effective connectivity between SMA and M1 underlie improved motor performance. PMID- 20005961 TI - Reduced hippocampal CA2, CA3, and dentate gyrus activity in asymptomatic people at genetic risk for Alzheimer's disease. AB - Previous functional magnetic resonance imaging (MRI) studies in healthy subjects with the apolipoprotein Eepsilon4 (APOE-4) genetic risk for Alzheimer's disease have shown increased activation during memory task performance in broadly distributed cortical regions. These findings have been hypothesized to reflect compensatory recruitment of intact brain regions that presumably result from subtle neural dysfunction reflecting incipient disease. In this study, we used high-resolution functional MRI in APOE-4 carriers and non-carriers to measure activity in hippocampal subregions (CA fields 1, 2, 3; dentate gyrus [DG], and subiculum) and adjacent medial temporal lobe (parahippocampal and entorhinal) subregions. We found reduced left CA2, CA3, and dentate gyrus (CA23DG) activity in cognitively intact APOE-4 carriers. These results suggest that reduced neural activity in hippocampal subregions may underlie the compensatory increase in extrahippocampal activity in people with a genetic risk for Alzheimer's disease prior to the onset of cognitive deficits. PMID- 20005963 TI - Bias in estimation of hippocampal atrophy using deformation-based morphometry arises from asymmetric global normalization: an illustration in ADNI 3 T MRI data. AB - Measurement of brain change due to neurodegenerative disease and treatment is one of the fundamental tasks of neuroimaging. Deformation-based morphometry (DBM) has been long recognized as an effective and sensitive tool for estimating the change in the volume of brain regions over time. This paper demonstrates that a straightforward application of DBM to estimate the change in the volume of the hippocampus can result in substantial bias, i.e., an overestimation of the rate of change in hippocampal volume. In ADNI data, this bias is manifested as a non zero intercept of the regression line fitted to the 6 and 12 month rates of hippocampal atrophy. The bias is further confirmed by applying DBM to repeat scans of subjects acquired on the same day. This bias appears to be the result of asymmetry in the interpolation of baseline and followup images during longitudinal image registration. Correcting this asymmetry leads to bias-free atrophy estimation. PMID- 20005964 TI - Early elevation of C-reactive protein correlates with severe infection and nonrelapse mortality in children undergoing allogeneic stem cell transplantation. AB - C-reactive protein (CRP) is an acute phase reactant that is a reliable marker of systemic inflammation and has been associated with increased morbidity and mortality following hematopoietic stem cell transplantation (HSCT) in adults. In this study, we evaluated whether early elevations of CRP were associated with various complications and nonrelapse mortality following HSCT in pediatric patients. Seventy pediatric patients had CRP levels drawn at regular time points during the first week following their transplants. Patients were followed for 100 days following transplant, and transplant-related complications were documented. Patients who subsequently developed severe infections had higher median CRP values than those without severe infections (median 8.03 mg/dL versus 1.64 mg/dL, P = .0008) as did those who suffered nonrelapse mortality compared with those who did not (12.6 mg/dL versus 2.44 mg/dL, P = .02). These findings suggest that elevated CRP values may be useful as a marker of individual pediatric patients with a higher risk for treatment-related morbidity and mortality. PMID- 20005965 TI - Gain of 1q21 is an unfavorable genetic prognostic factor for multiple myeloma patients treated with high-dose chemotherapy. AB - The prognostic significance of 1q21 gain, del(13)(q14), del(17)(p13), t(4;14)(p16.3;q32), and t(11;14)(q13;q32) detected by interphase fluorescein in situ hybridization (FISH) was studied in a cohort of 91 patients with newly diagnosed multiple myeloma (MM). 1q21 gain was detected in 37 of 91 patients (40.7%). In comparison with patients lacking 1q21 gain, patients with 1q21 gain had significantly shorter progression-free survival (PFS) (14.9 versus 27.4 months; P = .044) and worse 4-year overall survival (OS) (40.1% versus 76.2% of patients; P = <.001). PFS or OS were not influenced by the presence or absence of the other studied chromosomal abnormalities. Although the occurrence of 1q21 gain correlated with deletion of 13q14, the presence of 1q21 gain can be considered an independent prognostic factor, as no impact of del(13)(q14) as an isolated chromosomal abnormality on either PFS or OS has been observed. In comparison with patients lacking 1q21 gain, patients with 1q21 gain were significantly more likely to discontinue the preplanned treatment protocol because of disease progression or death. We conclude that 1q21 gain defines a prognostically unfavorable group of MM patients. PMID- 20005966 TI - Allogeneic stem cell transplantation for myelofibrosis with leukemic transformation. AB - Leukemic transformation (LT) from myelofibrosis has a very poor prognosis with the current treatment strategies. We hypothesized that allogeneic stem cell transplantation (ASCT) can improve outcomes for patients with LT, and reviewed 55 consecutive patients that were treated for myelofibrosis with ASCT at our institution. Fourteen patients (25%) were identified to have LT. Thirteen of these patients received induction chemotherapy and 6 achieved remission at the time of transplant. Conditioning regimen was melphalan (Mel)-based in 9 patients. All patients engrafted and achieved remission after transplant, whereas 4 subsequently relapsed. After a median follow-up of 31 months, 6 patients (49%) survived long term. Although limited by a small number of patients, this study suggests that patients with myelofibrosis and LT may achieve long-term remission after induction chemotherapy and ASCT. PMID- 20005967 TI - Response to tyrosine kinase inhibitor therapy in patients with chronic myelogenous leukemia relapsing in chronic and advanced phase following allogeneic hematopoietic stem cell transplantation. AB - Tyrosine kinase inhibitors (TKI) have been used to treat relapse of chronic myelogenous leukemia (CML) after allogeneic stem cell transplant (HSCT), with responses seen predominantly in chronic phase (CP) patients. This study aimed to analyze the response to TKI therapy and overall survival for patients relapsing predominantly in advanced phase. We retrospectively reviewed 22 patients treated with imatinib (n=20) and/or dasatinib (n=6) for relapsed CML after HSCT; 8 patients were in CP, and 14 patients had advanced disease. Seven patients also received donor lymphocyte infusions. Hematologic, cytogenetic, and molecular responses were analyzed. Nineteen patients (86%) achieved complete hematologic response (CHR), 17 patients (77%) achieved complete cytogenetic response (CCR), and 14 patients (64%) achieved complete molecular response (CMR). In advanced phase patients, 11 (79%) achieved CHR, 10 (71%) CCR, and 8 (57%) achieved CMR. Grade 3 or 4 cytopenias occurred in 10 cases. With median follow-up of 31.5 months from relapse, 14 (64%) patients remain alive, 13 in CMR. In multivariate analysis, the achievement of CMR was significantly correlated with OS with an odds ratio of 20.5 (95% confidence interval 2.3-182) P=.007. TKI therapy is capable of inducing durable molecular responses for CML relapsing after HSCT, both in chronic and advanced phases. The achievement of CMR appears to be crucial in providing long-term disease control for these patients. PMID- 20005968 TI - An assessment of the effect of human herpesvirus-6 replication on active cytomegalovirus infection after allogeneic stem cell transplantation. AB - Human herpesvirus-6 (HHV-6) may enhance cytomegalovirus (CMV) replication in allogeneic stem cell transplant (allo-SCT) recipients either through direct or indirect mechanisms. Definitive evidence supporting this hypothesis are lacking. We investigated the effect of HHV-6 replication on active CMV infection in 68 allo-SCT recipients. Analysis of plasma HHV-6 and CMV DNAemia was performed by real-time PCR. Enumeration of pp65 and IE-1 CMV-specific IFNgamma CD8(+) and CD4(+)T cells was performed by intracellular cytokine staining. HHV-6 DNAemia occurred in 39.8% of patients, and was significantly associated with subsequent CMV DNAemia in univariate (P=.01), but not in multivariate analysis (P=.65). The peak of HHV-6 DNAemia was not predictive of the development of CMV DNAemia. Timing and kinetics of active CMV infection were comparable in patients either with or without a preceding episode of HHV-6 DNAemia. The occurrence of HHV-6 DNAemia had no impact on CMV-specific T cell immunity reconstitution early after transplant. The receipt of a graft from an HLA-mismatched donor was independently associated with HHV-6 (P=.009) and CMV reactivation (P=.04). The data favor the hypothesis that a state of severe immunosuppression leads to HHV-6 and CMV coactivation, but argue against a role of HHV-6 in predisposing to the development of CMV DNAemia or influencing the course of active CMV infection. PMID- 20005969 TI - Calnexin phosphorylation: linking cytoplasmic signalling to endoplasmic reticulum lumenal functions. AB - Calnexin is an abundant integral membrane phosphoprotein of the endoplasmic reticulum (ER) of eukaryotic cells. The role of the luminal domain as an N glycoprotein specific lectin has been well-established. Cytosolic C-terminal domain phosphorylation of calnexin has recently been elucidated in glycoprotein folding and quality control. Signalling of the presence of unfolded proteins from the lumen of the ER is mediated by the three ER membrane sensor proteins Ire1, ATF6 and PERK. The observation that the C-terminus of calnexin is differentially phosphorylated when glycoproteins are misfolded initiated our search for functional roles of calnexin phosphorylation. Recent studies have defined a role for phosphorylation at a proline-directed kinase site (Ser563) in ER protein quality control, while phosphorylation at a casein kinase 2 site (Ser534, Ser544) may be linked to transport functions. There are also four other abundant integral membrane phosphoproteins in the ER, and these may be components of other signalling pathways that link and coordinate other ER functions with the rest of the cell. PMID- 20005970 TI - Dietary nitrite ameliorates renal injury in L-NAME-induced hypertensive rats. AB - Nitric oxide (NO) has numerous important functions in the kidney, and long-term blockage of nitric oxide synthases in rats by L-NAME results in severe hypertension and progressive kidney damage. On the other hand, NO production seems to be low in patients with chronic kidney disease (CKD), and NO deficiency may play a role in CKD progression. In this review, we summarized the mechanisms of amelioration of renal injury induced by L-NAME treated rats by treatment of nitrite. First, we demonstrate whether orally-administrated nitrite-derived NO can shift to the circulation. When 3mg/kg body weight Na(15)NO(2) was orally administered to rats, an apparent EPR signal derived from Hb(15)NO (A(z)=23.4 gauss) appeared in the blood, indicating that orally ingested nitrite can be a source of NO in vivo. Next, in order to clarify the capacity of nitrite to prevent renal disease, we administered low-dose nitrite (LDN: 0.1mg of sodium nitrite in 1L of drinking water), medium-dose nitrite (MDN: 1mg sodium nitrite/L, which corresponds to the amount of nitrite ingested by vegetarians), or high-dose nitrite (HDN: 10mg sodium nitrite/L) to rats simultaneously with L-NAME (1 g l NAME/L) for 8 weeks, then examined the blood NO level as a hemoglobin-NO adduct (iron-nitrosyl-hemoglobin) using electron paramagnetic resonance spectroscopy, urinary protein excretion, and renal histological changes at the end of the experiment. It was found that oral administration of MDN and HDN but not LDN increased the blood iron-nitrosyl-hemoglobin concentration to the normal level, ameliorated the L-NAME-induced proteinuria, and reduced renal histological damage. The findings demonstrate that chronic administration of a mid-level dietary dose of nitrite restores the circulating iron-nitrosyl-hemoglobin levels reduced by L-NAME and that maintenance of the circulating iron-nitrosyl hemoglobin level in a controlled range protects against L-NAME-induced renal injury. Taking these findings together, we propose that dietary supplementation of nitrite is a potentially useful nonpharmacological strategy for maintaining circulating NO level in order to prevent or slow the progression of renal disease. It had been believed that nitrite could result in intragastric formation of nitrosamines, which had been linked to esophageal and other gastrointestinal cancers. However, there is no positive association between the intake of nitrate or nitrite and gastric and pancreatic cancer by recent researches. Furthermore, nitrate-derived NO formation pathway is a possible mechanism for the hypotensive effect of vegetable- and fruit-rich diets, which may explain, at least in part, the mechanism of the Dietary Approach to Stop Hypertension (DASH) diet-induced hypotensive and organ-protective effects. Further research is needed to investigate the interaction between nitrite-nitrate intakes and human health. PMID- 20005971 TI - The early history of the Sox genes. AB - The Sox gene family was discovered in 1990 through the cloning of Sry, the mammalian Y-linked testis-determining gene. This was long before genome sequencing was routine, but it was an exciting time when rapid progress was being made in gene discovery, through positional and functional cloning techniques, and via homology within and between species. It had become clear that many genes were members of often quite substantial families, related by the possession of similar domains within their coding sequences and corresponding proteins. Members of the SOX protein family were subsequently shown to have many diverse roles during development in metazoan species, but are all related by a similar HMG box type of DNA binding domain, which gives them functional properties distinct from other transcription factors. This is a brief account of how the first Sox genes were found. PMID- 20005972 TI - SRY: A transcriptional activator of mammalian testis determination. AB - Sry (sex-determining region Y) is the sex-determining gene on the mammalian Y chromosome, which encodes a transcription factor containing a DNA-binding domain characteristic of some high mobility group proteins (HMG box). It is the founder member of the Sox (Sry-related HMG box) gene family and is therefore classified in the Sox A group. In mice, the transient expression of Sry between 10.5 and 12.5 dpc triggers the differentiation of Sertoli cells from the supporting cell precursor lineage, which would otherwise give rise to granulosa cells in ovaries. However, little was known about the target genes of SRY and molecular mechanisms how SRY leads to testis development. Recent work has provided evidence that SRY binds directly to a testis-specific enhancer of Sox9 (TES) and activates Sox9 expression in co-operation with steroidogenic factor 1 (SF1). Furthermore, this SRY action is limited to a certain time period during embryogenesis. PMID- 20005974 TI - Electronegative LDL induction of apoptosis in macrophages: involvement of Nrf2. AB - The aim of this study was to determine the apoptotic pathways and mechanisms involved in electronegative LDL [LDL(-)]-induced apoptosis in RAW 264.7 macrophages and the role of Nrf2 in this process. Incubation of RAW 264.7 macrophages with LDL(-) for 24 h resulted in dose-dependent cell death. Activated caspases were shown to be involved in the apoptosis induced by LDL(-); incubation with the broad caspase inhibitor z-VAD prevented apoptosis in LDL(-)-treated cells. CD95 (Fas), CD95 ligand (FasL), CD36 and the tumor necrosis factor (TNF) ligand Tnfsf10 were overexpressed in LDL(-)-treated cells. However, Bax, Bcl-2 and Mcl-1 protein levels remained unchanged after LDL(-) treatment. LDL(-) promoted hyperpolarization of the mitochondrial membrane, elevated reactive oxygen species (ROS) production and translocation of Nrf2 to the nucleus, a process absent in cells treated with native LDL. Elicited peritoneal macrophages from Nrf2-deficient mice exhibited an elevated apoptotic response after challenge with LDL(-), together with an increase in the production of ROS in the absence of alterations in CD36 expression. These results provide evidence that CD36 expression induced by LDL(-) is Nrf2-dependent. Also, it was demonstrated that Nrf2 acts as a compensatory mechanism of LDL(-)-induced apoptosis in macrophages. PMID- 20005973 TI - Proteomics identifies multipotent and low oncogenic risk stem cells of the spleen. AB - The adult spleen harbors a population of naturally occurring multipotent stem cells of non-lymphoid lineage (CD45-). In animal models, these splenic stem cells can directly or indirectly contribute to regeneration of bone, inner ear, cranial nerves, islets, hearts and salivary glands. Here we characterize the CD45- stem cell proteome to determine its potential broader multipotency versus its protection from malignant transformation. Using state-of-the-art proteomics and in vivo testing, we performed functional analyses of unique proteins of CD45- (non-lymphoid) splenic stem cells, as compared with CD45+ (lymphoid) cells. CD45- stem cell-specific proteins were identical to those in iPS, including OCT3/4, SOX2, KLF4, c-MYC and NANOG. They also expressed Hox11, Gli3, Wnt2, and Adam12, the benchmark transcription factors of embryonic stem cells. These transcription factors were functional because their mRNA was upregulated in the spleen in association with ongoing damage to the pancreas and salivary glands, organs to which they normally contribute stem cells. We also show low likelihood of malignant transformation. Our proteomic and functional analyses reveals that naturally occurring CD45- stem cells of the spleen are the first-ever candidates for naturally occurring population of embryonic and iPS cells with low oncogenic risk. Given their presence in normal humans and mice, splenic stem cells are poised for translational research. PMID- 20005975 TI - Effect of opioid compounds on feeding and activity of the cockroach, Periplaneta americana. AB - Opioid peptides have been implicated in regulation of feeding in invertebrates. Studies have suggested that receptors for opioids are present in cockroaches and that these receptors play roles in affecting both behaviour and feeding. We examined the effect of micro, delta, and kappa opioid receptor agonists and antagonists on feeding, mass changes and activity in the cockroach, Periplaneta americana. The kappa antagonist, nor-binaltorphimine, significantly increased food intake, while naltrexone (general antagonist) and naloxonazine (micro antagonist) both reduced feeding. A large mass loss was observed in cockroaches treated with nor-binaltorphimine, despite the increased food intake. Males did not lose as much mass during the 3h as females, although drug treatment did have some effect on the loss. Time of activity (%) was not influenced by any drug. Water loss experiments suggested that nor-binaltorphimine increased water loss, accounting for the mass loss despite the increased feeding. We suggest that two populations of opioid receptors are present as previously reported, with one affecting feeding and the other involved with evaporative water loss. PMID- 20005977 TI - Increased risk for non-melanoma skin cancer in patients with inflammatory bowel disease. AB - BACKGROUND & AIMS: Patients with inflammatory bowel disease (IBD) might be at increased risk for certain malignancies. We evaluated the risk of non-melanoma skin cancer (NMSC) in patients with IBD and determined how immunosuppressive and biologic medications affect this risk. METHODS: We performed retrospective cohort and nested case-control studies by using administrative data from PharMetrics Patient Centric Database. In the cohort study, 26,403 patients with Crohn's disease (CD) and 26,974 patients with ulcerative colitis (UC) were each matched to 3 non-IBD controls. NMSC risk was evaluated by incidence rate ratio (IRR). In the nested case-control study, 387 CD patients and 355 UC patients with NMSC were each matched to 4 IBD patients without NMSC by using incidence density sampling. Conditional logistic regression was used to determine the association between specific IBD medication use and NMSC. RESULTS: In the cohort study, the incidence of NMSC was higher among patients with IBD compared with controls (IRR, 1.64; 95% confidence interval [CI], 1.51-1.78). In the nested-case control study, recent thiopurine use (< or =90 days) was associated with NMSC (adjusted odds ratio [OR], 3.56; 95% CI, 2.81-4.50), as was recent biologic use among patients with CD (adjusted OR, 2.07; 95% CI, 1.28-3.33). Persistent thiopurine use (>365 days) was associated with NMSC (adjusted OR, 4.27; 95% CI, 3.08-5.92), as was persistent biologic use among patients with CD (adjusted OR, 2.18; 95% CI, 1.07-4.46). CONCLUSIONS: Patients with IBD, especially those who receive thiopurines, are at risk for NMSC. Appropriate counseling and monitoring of such patients with IBD are recommended. PMID- 20005976 TI - Signaling mechanisms in thyroid hormone-induced cardiac hypertrophy. AB - Cardiac hypertrophy is a significant independent risk factor for increased mortality, comprising of maladaptive changes in cellular, molecular and metabolic processes that ultimately lead to heart failure. However, cardiac hypertrophy represents a continuum from physiological to compensatory to pathological hypertrophy, so that treatment modalities aimed to shift hypertrophy towards the physiological phenotype would represent an attractive therapeutic strategy. Many of the physiological changes caused by thyroid hormone (TH) treatment may provide direct benefit to the failing heart. Recent experimental studies have shown that TH rapidly activates pro-survival PKB/Akt-mTOR signaling pathways, thus providing cytoprotection and increasing synthesis of normal contractile proteins and metabolic enzymes. TH induces a normal physiological phenotype by binding to nuclear TH receptors that regulate expression of specific genes which promote cell survival and enhance contractile function. Physiological cardiac growth occurs with a coordinated angiogenic response that normalizes myocardial perfusion during hypertrophy, and recent studies support a significant role for TH and its endothelial cell surface integrin receptors and nuclear receptors in neovascularization during TH-induced hypertrophy. The present review examines these molecular mechanisms and intracellular signaling pathways activated in thyroid hormone-induced cardiac hypertrophy that support its therapeutic potential in the treatment of heart disease. PMID- 20005978 TI - Stool polymerase chain reaction for Helicobacter pylori detection and clarithromycin susceptibility testing in children. AB - BACKGROUND & AIMS: This study was undertaken in a pediatric gastroenterology clinic to retrospectively evaluate a real-time polymerase chain reaction (PCR) for the detection and clarithromycin susceptibility testing of Helicobacter pylori using stool specimens. METHODS: All consecutive children who underwent a gastroscopy between March 2006 and February 2009 and also having been examined by stool PCR were enrolled. Rapid urease test, histology, and culture were the reference methods for the detection of H pylori and E-test for susceptibility testing, respectively. RESULTS: A total of 143 children (mean age, 10.8 y; range, 2.8-17.9; males:females, 1:1.5) were evaluable. Sensitivity, specificity, and test accuracy for the detection of H pylori were 83.8%, 98.4%, and 90.2%, respectively. Sensitivity, specificity, and accuracy for the detection of clarithromycin resistance were 89.2%, 100%, and 94.0%, respectively. CONCLUSIONS: Stool PCR was a reliable and useful noninvasive tool for detection and clarithromycin susceptibility testing of H pylori in a pediatric population with a high prevalence of clarithromycin-resistant strains. PMID- 20005979 TI - Disconnected pancreatic duct with pancreas necrosis, treated with transgastric debridement and pancreatic duct stent. PMID- 20005980 TI - Acute pancreatitis part II: approach to follow-up. PMID- 20005981 TI - Medical economies 2009. PMID- 20005983 TI - Electronic image of the month. Adenosquamous carcinoma of the pancreas. PMID- 20005982 TI - Adalimumab level in breast milk of a nursing mother. PMID- 20005984 TI - Personalized dose reduction for computed tomography scanning: size matters, so does prior radiation exposures. PMID- 20005985 TI - Artesunate protects sepsis model mice challenged with Staphylococcus aureus by decreasing TNF-alpha release via inhibition TLR2 and Nod2 mRNA expressions and transcription factor NF-kappaB activation. AB - Gram-positive bacteria have become the most common organisms responsible for the development of sepsis. Staphylococcus aureus (S. aureus) is the major gram positive pathogen in both community-acquired and nosocomial infections. The Mortality associated with nosocomial infections caused by S. aureus may vary but are generally high. In the present study, we found that artesunate (AS) could protect mice against a lethal challenge with heat-killed S. aureus in a dose dependent manner, and AS in combination with ampicillin sodium-sulbactam sodium (AMPS) could further increase survival of mice challenged with live S. aureus than AMPS alone. This protection was associated with reductions of serum at TNF alpha level. In in vitro experiments, AS-pretreatment strongly inhibited TNF alpha release from murine peritoneal macrophage induced by heat-killed S. aureus or peptidoglycan in a dose-dependent manner. AS reduced the Toll like receptor 2 (TLR2) and nucleotide-binding oligomerization domain containing 2 (Nod2) mRNA expressions up-regulated by heat-killed S. aureus and inhibited NF-kappaB activation induced by heat-killed S. aureus. In conclusion, our results demonstrated that AS-mediated protection on septic mice challenged with S. aureus was associated with its reduction on TNF-alpha release via inhibition of TLR2 and Nod2 mRNA expressions and transcription factor NF-kappaB activation. PMID- 20005987 TI - Uncoupling protein-2 and cancer. AB - Cancer cells respond to unfavorable microenvironments such as nutrient limitation, hypoxia, oxidative stress, and host defense by comprehensive metabolic reprogramming. Mitochondria are linked to this complex adaptive response and emerging evidence indicates that uncoupling protein-2 (UCP2), a mitochondrial inner membrane anion carrier, may contribute to this process. Effects of UCP2 on mitochondrial bioenergetics, redox homeostasis, and oxidant production in cancer cells may modulate molecular pathways of macromolecular biosynthesis, antioxidant defense, apoptosis, cell growth and proliferation, enhancing robustness and promoting chemoresistance. Elucidation of these interactions may identify novel anti-cancer strategies. PMID- 20005986 TI - Oligodendroglial differentiation induces mitochondrial genes and inhibition of mitochondrial function represses oligodendroglial differentiation. AB - Demyelination occurs in multiple inherited mitochondrial diseases. We studied which genes were induced as a consequence of differentiation in rodent and human oligodendroglia. Cholesterol, myelin and mitochondrial genes were significantly increased with oligodendroglial differentiation. Mitochondrial DNA content per cell and acetyl CoA-related transcripts increased significantly; thus, the large buildup of cholesterol necessary for myelination appears to require mitochondrial production of acetyl-CoA. Oligodendroglia were treated with low doses of the mitochondrial inhibitor rotenone to test the dependence of differentiation on mitochondrial function. Undifferentiated cells were resistant to rotenone, whereas differentiating cells were much more sensitive. Very low doses of rotenone that did not affect viability or ATP synthesis still inhibited differentiation, as measured by reduced levels of the myelin transcripts 2',3' Cyclic Nucleotide-3'-Phosphodiesterase and Myelin Basic Protein. Thus, mitochondrial transcripts and mtDNA are amplified during oligodendroglial differentiation, and differentiating oligodendroglia are especially sensitive to mitochondrial inhibition, suggesting mechanisms for demyelination observed in mitochondrial disease. PMID- 20005988 TI - Gases in the mitochondria. AB - Gasomodulators - nitric oxide, carbon monoxide and hydrogen sulphide - are important physiological mediators that have been implicated in disorders such as neurodegeneration and sepsis. Some of their biological functions involve the mitochondria. In particular, their inhibition of cytochrome c oxidase has received much attention as this can cause energy depletion and cytotoxicity. However, reports that cellular energy production and cell survival are maintained even in the presence of gasomodulators are not uncommon. In both cases, modulation of mitochondrial targets by the gasomodulators appears to be an important event. We provide an overview of the effects of the gasomodulators on the mitochondria. PMID- 20005989 TI - Effects of alpha-mangostin on mitochondrial energetic metabolism. AB - Although alpha-mangostin prevents from toxicity associated to oxidative stress, it also promotes apoptotic cell death in cancer cells. Such effects have been associated with mitochondrial membrane depolarization and cytochrome c release. Therefore, the aim of this work was to analyze the potentially harmful effect of this natural compound on relevant parameters of mitochondrial function from normal tissue. Our results showed that alpha-mangostin protected mitochondria from peroxidative damage, but at high concentration, it acted as an uncoupler, reduced dramatically ADP-stimulated respiration and inhibited the activity of respiratory complex IV, making mitochondria prone to permeability transition, which is a mitochondrial player on cell fate. PMID- 20005991 TI - ECG and VT/VF symposium. PMID- 20005992 TI - QT dynamicity, microvolt T-wave alternans, and heart rate variability during 24 hour ambulatory electrocardiogram monitoring in the healthy newborn of first to fourth day of life. AB - BACKGROUND: Twenty-four hour ambulatory electrocardiogram (AECG) monitoring is an established technique for integrated assessment of heart rhythm; however, comprehensive description of serial changes in cardiac electrophysiology over the first days of life in humans is lacking. The aim of this study was to determine the patterns of circadian heart rhythm based on AECG evaluation in newborns. METHODS: Twenty healthy newborns (14 boys and 6 girls) were serially examined with AECG at days 1, 2, and 4 after birth. Heart rate (HR), arrhythmias, QT dynamicity, microvolt T-wave alternans, and various indices of HR variability (HRV) including deceleration/acceleration capacity analysis were analyzed. RESULTS: There were no sex differences in HR. Supraventricular premature beats were noted in 35%, ventricular-in 15 % of newborns. Slope QT/RR was 0.35 (0.3 0.5); intercept QT/RR was 124 (93-148), QT/RR correlation coefficient (r) was 0.63 (0.53-0.85). Peak value of T-wave alternans was 32 +/- 8 (12-55) muV. Low level of HRV was typical for all parameters of time-domain analysis compared with normal limits for older children. The overall mean values of deceleration/acceleration capacity were 3.38 +/- 0.57 (2.16-4.13) and -3.58 +/- 0.67 (-2.13 to -4.38) milliseconds, respectively. CONCLUSION: The healthy newborns exhibit peculiarities of 24-hour cardiac rhythm with isolated premature beats, pauses of sinus rhythm less 1000 milliseconds, steep slope of QT/RR by analysis of QT dynamicity. There are low HRV, and symmetrical AC/DC capacity was typically for autonomic regulation of HR, probably due to high sympathetic activity at this age. PMID- 20005993 TI - Two automatic QT algorithms compared with manual measurement in identification of long QT syndrome. AB - BACKGROUND: Long QT syndrome (LQTS) is an inherited disorder that increases the risk of syncope and malignant ventricular arrhythmias, which may result in sudden death. METHODS: We compared manual measurement by 4 observers (QT(manual)) and 3 computerized measurements for QT interval accuracy in the diagnosis of LQTS: 1. QT measured from the vector magnitude calculated from the 3 averaged orthogonal leads X, Y, and Z (QTVCG) and classified using the same predefined QTc cut-points for classification of QT prolongation as in manual measurements; 2. QT measured by a 12-lead electrocardiogram (ECG) program (QTECG) and subsequently classified using the same cut-points as in (1) above; 3. The same QT value as in (2) above, automatically classified by a 12-lead ECG program with thresholds for QT prolongation adjusted for age and sex (QTinterpret). The population consisted of 94 genetically confirmed carriers of KCNQ1 (LQT1) and KCNH2 (LQT2) mutations and a combined control group of 28 genetically confirmed noncarriers and 66 unrelated healthy volunteers. RESULTS: QT(VCG) provided the best combination of sensitivity (89%) and specificity (90%) in diagnosing LQTS, with 0.948 as the area under the receiver operating characteristic curve. The evaluation of QT measurement by the 4 observers revealed a high interreader variability, and only 1 of 4 observers showed acceptable level of agreement in LQTS mutation carrier identification (kappa coefficient >0.75). CONCLUSION: Automatic QT measurement by the Mida1000/CoroNet system (Ortivus AB, Danderyd, Sweden) is an accurate, efficient, and easily applied method for initial screening for LQTS. PMID- 20005994 TI - Concomitant left anterior descending coronary artery and right coronary artery occlusion with typical anterior ST depression and inferior ST elevation. AB - We report the case of a 41-year-old man with acute myocardial infarction showing first ST elevation in V1-V6-DI-aVL leads followed by a typical V2-V4 ST depression (concomitant occlusion of proximal diagonal branch with an incomplete left anterior descending occlusion) and DII-DIII-aVF ST elevation. At coronary angiography, a proximal left anterior descending coronary stenosis with right coronary artery thrombosis was found. PMID- 20005995 TI - Liver transplant for relapsed undifferentiated embryonal sarcoma in a young child. AB - Undifferentiated embryonal sarcoma of the liver is a rare hepatic malignancy of childhood with a historically poor prognosis. Recent improvements in outcomes have been reported in small numbers of cases with the use of combination therapy involving aggressive surgical resection and chemotherapy. Complete surgical resection is frequently difficult to achieve when the location of the tumor is along the margins of the major hepatic vessels (portal vein, hepatic vein, and hepatic artery). Here we report a case of undifferentiated embryonal sarcoma of the liver that recurred along surgical hepatic vein margins in a 9-year-old boy who subsequently underwent orthotopic liver transplantation from a cadaveric donor. The patient has been in continuous clinical remission for the last 5 years. PMID- 20005996 TI - Ectopic nephrogenic rests in children: the clinicosurgical implications. AB - INTRODUCTION/BACKGROUND: Ectopic nephrogenic rests (ENR) are rare. The incidental discovery of these lesions in children has particular clinicosurgical implications, especially given the association between ENR and the development of extrarenal Wilms' tumors (ERWT). METHODOLOGY: We reviewed the hospital records of patients with ERWT and ENR treated at our hospital over a 10-year period to identify those patients with histopathologically confirmed ENR and/or ERWT. RESULTS: Ninety-five children with Wilms' tumor (WT) were identified, but only 1 case of ENR and ERWT. This patient was a 14-month-old boy who was incidentally found to have a mass in the left inguinal canal during orchiopexy. After histology, a provisional diagnosis of ENR was made. Six months later, the child went on to develop an ERWT at the same site. Periodic postsurgical follow-up has been uneventful. DISCUSSION AND CONCLUSIONS: This was the only case of ENR and ERWT in child in a 10-year review of patients with WT at our hospital. Our experience stresses the importance of including ENR in any working differential diagnosis of unexpected masses in the inguinal canal in children and underscores why careful long-term follow-up is mandatory. The reasons for the malignant transformation of ENR into primary ERWT are unknown, but our experience lends support for the theory that ENR are precursor lesions to the development of WT even in ectopic sites. The case also provides an example of the kind of technical difficulties presented by paratesticular masses during laparoscopy. PMID- 20005997 TI - Colocolic intussusception in a preterm infant with intestinal malrotation. AB - The association of intussusception and intestinal malrotation is referred to as Waugh syndrome. A prospective study among 49 children with intussusception found a 40% incidence of malrotation. We describe the rare occurrence of colocolic intussusception in a neonate with malrotated intestine. We believe this is the first reported case in the neonatal period where the pathogenic lead point was an intestinal lymphangioma. PMID- 20005998 TI - Shoulder dislocation in a very-low-birth-weight infant: case report and review of the literature. AB - Shoulder dislocation in a preterm infant is very rare. We report a case of a preterm infant, with gestational age of 25 2/7 weeks and birth weight of 910 g, with anterior shoulder dislocation after breech delivery. The infant was successfully treated with closed reduction. In general, long-term outcome of shoulder dislocation in these infants is good. However, prompt recognition and treatment of the dislocated shoulder are essential to maintain normal shoulder girdle function and to prevent functional disability. PMID- 20005999 TI - A unique case of a congenital diaphragmatic hernia in a boy with albinism. AB - Congenital diaphragmatic hernia and oculocutaneous albinism are both rare birth defects that can be diagnosed in the newborn period. However, they have not been previously reported to have occurred together. This report describes a unique case of a male Asian baby with oculocutaneous albinism and a right-sided congenital diaphragmatic hernia. PMID- 20006000 TI - Use of gauze-based negative pressure wound therapy in a pediatric burn patient. AB - Negative pressure wound therapy (NPWT) is described as it is used in the treatment of an infant burn victim. This case highlights the ability and techniques used to maintain an airtight dressing seal in the perirectal region. Use of this dressing type post-skin grafting allowed for 100% graft adhesion and no bacterial contamination despite close proximity to the rectum. Favorable experience and outcome with this patient are strong indicators that NPWT should be considered as a viable treatment in pediatric populations and that situations where body contour or fluids may make NPWT difficult to administer should not be a deterrent to therapy. PMID- 20006001 TI - Intradiaphragmatic extralobar sequestration-a rare pulmonary anomaly. AB - Extralobar pulmonary sequestrations are most commonly found within the thoracic cavity, but have been described within the abdomen. We present the case of a 16 month-old boy with an intradiaphragmatic pulmonary sequestration and demonstrate a computed tomographic scan finding that might help identify this extremely rare abnormality preoperatively. PMID- 20006002 TI - Long-term survival and renal transplantation in a monozygotic twin with cloacal dysgenesis sequence. AB - Cloacal dysgenesis sequence (CDS) is a severe hindgut malformation occurring in 1:50,000 to 250,000 live births (Qureshi et al. Prenatal diagnosis of cloacal dysgenesis sequence: differential diagnosis from other forms of fetal obstructive uropathy. Fetal Diagn Ther 1998;13:69-74; Bargaje et al. Cloacal dysgenesis sequence. Ann Diagn Pathol 2008;12:62-66). It is characterized by a smooth perineum with no urethral, vaginal, or anal openings, and lack of labioscrotal development. Typically, the bladder, vagina, and colon each end blindly, although persistent cloaca without perineal orifice can be seen. With no egress for urine, infants have renal insufficiency and pulmonary hypoplasia, usually making CDS lethal (Sahinoglu Z et al. The prenatal diagnosis of cloacal dysgenesis in six cases: can the termination of pregnancy always be the first choice? Prenat Diagn 2004;24:10-16). Reported survivors have had a persistent urachus or have been spared the effects of oligohydramnios by the presence of a twin (Liang X. Cloacal dysgenesis sequence: observations in four patients, including three fetuses of second trimester gestation. Pediatr Dev Pathol 1998;1:281-288). We report a case of long-term survival, currently to 25 months of age, and renal transplantation in a monochorionic, diamniotic twin girl with CDS. PMID- 20006003 TI - Renal cell carcinoma originating in a renal cyst in a 12-year-old girl. AB - We report a 12-year-old girl with renal cell carcinoma originating in a cyst of the left kidney. Ultrasonography revealed 2 small hyperechoic masses in the luminal side of a cyst. Although hypervascularity was not detected in the cyst by computed tomography, the possibility of malignancy could not be ruled out because of the presence of 2 solid masses. Therefore, partial left nephrectomy was performed. On histopathologic examination, the 2 solid masses within the cyst were found to be renal cell carcinoma. This patient remains disease-free at 4 years after partial nephrectomy. PMID- 20006004 TI - Mechanical compression with secondary ischemia as a possible cause of atresias associated with omphalocele. AB - We report 2 cases of omphalocele associated with intestinal atresia. In the context of this unusual sequence, we discuss the etiopathogenesis of intestinal atresias in association with tight abdominal wall defects. PMID- 20006005 TI - Risks and benefits of ending of mass screening for neuroblastoma at 6 months of age in Japan. AB - PURPOSE: The mass screening (MS) for neuroblastoma (NB) at 6 months of age in Japan was discontinued in 2004. This study assessed the risks and benefits of MS based on an analysis of NB detected before or after discontinuation of MS in Japan. METHODS: The clinical features and Brodeur's genetic type based on MYCN, DNA ploidy, and other genetic aberrations were assessed in 113 NB patients (20 cases after and 93 cases [55 MS cases] before the discontinuation of MS) older than 6 months treated at one institution since 1985. RESULTS: The 20 patients with NBs detected after MS was discontinued ranged in age from 7 to 67 months, 12 patients were stage 4, and 11 patients would have been detected at 6 months of age if they had undergone MS. The Brodeur's genetic type of these 20 patients showed that 30% (6/20) were type 1 (low risk), 55% (11/20) were type 2A (intermediate risk), and 15% (3/20) were type 2B (high risk). Of 93 patients with NB detected before MS was discontinued, 60% (56/93) were type 1, 18% (17/93) were type 2A, and 22% (20/93) were type 2B. Among the type 2A patients, 82% (9/11) of the patients detected after MS was discontinued showed stage 4, whereas only 50% (9/18) of those diagnosed before MS was discontinued were stage 4. The genetic analysis using single nucleotide polymorphism (SNP) array for type 2A showed that the pattern of genetic aberration was equivalent in those detected either before or after MS was discontinued. CONCLUSIONS: There was a decrease of type 1 and an increase of type 2A NB in patients after MS was discontinued in Japan. These results suggest that most of the type 1 detected by MS has regressed, and most of the type 2A detected by MS has appeared sporadically as advanced NB in patients older than 1 year. PMID- 20006006 TI - Telomere biology in neuroblastoma: telomere binding proteins and alternative strengthening of telomeres. AB - PURPOSE: Neuroblastoma (NBL) shows remarkable biologic heterogeneity, resulting in favorable or unfavorable prognoses. Previously, we reported that most unfavorable NBLs express high telomerase activity to maintain telomere length. Recently, telomere binding proteins (TBPs) and alternative lengthening of telomeres (ALTs) have been identified as key factors of telomere maintenance. METHODS: To evaluate the correlation between telomerase activity, telomere length, and the expression levels of TBPs in NBL, we analyzed and quantified these factors in 121 untreated NBLs. RESULTS: Shortened and elongated telomeres were detected in 21 (17.3%) and 11 cases (9.0%), respectively, and there was a significant correlation between telomere length and the length of the 3' overhang. The tumors with shortened or elongated telomeres showed significant lower expression of TBPs, except for RAP1. Although telomerase activity did not correlate with telomere length, 16 of 22 cases with high telomerase activity and 5 of 9 cases (ALT tumors) that showed long telomeres without high telomerase activity resulted in death. High-dose chemotherapy did not have much effect on these deceased ALT cases, but their survival periods were more than 2 years and relatively long compared with the deceased cases with nonelongated telomeres, suggesting that chemoresistance in ALT tumors may be related to slow growth rates. CONCLUSIONS: High telomerase activity is a poor prognostic factor in NBL. In the cases without high telomerase activity, those with elongated telomere also showed poor outcomes because of chemoresistance. Therefore, ALT and TBPs may be biomarkers for chemosensitivity in NBL. Thus, a better understanding of telomere biology may help define the characteristics of individual NBLs. PMID- 20006007 TI - Childhood pancreatic tumors: a single institution experience. AB - PURPOSE: The rarity and histopathologic diversity of primary pancreatic neoplasms in children have made it difficult to predict prognosis and to develop optimal management protocols. METHODS: A 90-year (1918-2007), single institution, retrospective review of all patients with neoplastic pancreatic masses was performed. RESULTS: Eighteen patients were identified with 7 distinct histopathologic subtypes. The most common were gastroenteropancreatic neuroendocrine, solid pseudopapillary, and acinar tumors. There were 6 benign and 12 malignant tumors. Six patients had disease outside the pancreas at their initial operation. There were 7 deaths (41%), 2 related to the initial operation, 3 from disease progression, 1 from a small bowel obstruction, and 1 from necrotizing pancreatitis. Five deaths were in patients with extrapancreatic disease found at initial operation. The median duration of follow-up for the 10 survivors was 41 months. CONCLUSION: In adults, pancreatic ductal adenocarcinoma is by far the most common histopathologic subtype, with other subtypes more common in children. Stage is an important prognostic factor. Long-term disease free survival in childhood pancreatic malignancies is achievable with complete surgical resection, prognosis, and adjuvant treatment, depending on the histopathologic type. PMID- 20006008 TI - Triage of pediatric injuries after the 2008 Wen-Chuan earthquake in China. AB - PURPOSE: The study aimed to review the effect of modifying triage strategies on the consultation and operation waiting times during the Wen-Chuan earthquake in China in 2008. METHOD: The triage during the post-earthquake period was categorized into 3 phases. The consultation and operation waiting times were analyzed. RESULTS: Of the 119 admitted children, there were 58 boys and 61 girls. Most of the victims were school-aged. In phase 1 (24 hours after the quake), the triage waiting time was 78 minutes. The waiting time for pediatric subspecialty consultation was 3.5 hours. There was an additional 7.5-hour delay before operation. In phase 2 (24-72 hours after the quake), senior pediatric surgeons carried out the triage and consultation. The consultation waiting time was reduced to 31 minutes. Four rotating teams operated 24 hours a day. The waiting time for operation was reduced to 4.5 hours. In phase 3 (4-19 days after the earthquake), gas gangrene screening was implemented. The triage waiting times for closed and open injuries were 47 and 64 minutes, respectively. Operation waiting times of 4.4 and 4.8 hours were recorded for closed and open injuries, respectively. Compared to that of phase 1, the waiting times for both consultation and operation of phases 2 and 3 were significantly shortened (P < .05). Most of the (89%) of the injuries were orthopedic traumas with lower limb fracture being the most common injury. Intraabdominal and thoracic injuries were relatively uncommon. CONCLUSIONS: (1) Triage by pediatric surgeons in the reception area greatly reduced the delay of treatment and (2) the predominance of orthopedic injuries resulting from the earthquake indicates the focus of medical resource allocation in natural disasters of this type in the future. PMID- 20006009 TI - Esophageal atresia/tracheoesophageal fistula in very low-birth-weight neonates: improved outcomes with staged repair. AB - INTRODUCTION: The surgical management of esophageal atresia with distal tracheoesophageal fistula (EA/TEF) involves early division of the TEF and primary esophageal anastomosis. However, in premature infants, the morbidity associated with primary repair remains high, and the optimal surgical approach has not been well defined. METHODS: Medical records of very low-birth-weight infants (<1500 g) with EA/TEF from June 1987 to 2008 were retrospectively reviewed. Patients were separated into 2 groups: (1) primary repair and (2) ligation and division of TEF followed by delayed repair of EA. Demographics, anastomotic, and postoperative complications were compared. RESULTS: Twenty-five premature infants with EA/TEF were identified. Sixteen patients (64%) underwent primary repair, and 9 (36%) were repaired in a staged manner. The leak rate confirmed by esophagram was significantly higher after primary repair (50%) compared to staged repair (0%) (P = .034). Strictures occurred significantly more often in the primary repair (81%) vs the staged repair (33%) group (P = .036). Postoperative pneumonia and sepsis were significantly higher in patients treated with primary repair (P = .028). CONCLUSION: Staged repair of EA/TEF in very low-birth-weight premature infants results in a significantly lower rate of anastomotic complications and overall morbidity and should be considered the preferred surgical approach in this group of patients. PMID- 20006010 TI - Influence of thoracoscopic esophageal atresia repair on esophageal motor function and gastroesophageal reflux. AB - BACKGROUND/PURPOSE: Thoracoscopic repair has recently been attempted in newborns with esophageal atresia (EA), but it remains unclear whether thoracoscopic dissection reduces pathological gastroesophageal reflux. We investigated the influence of a thoracoscopic approach on esophageal motor function in patients with EA. METHODS: Clinical and gastrointestinal data of 10 patients with EA with open repair (group A) and 7 with thoracoscopic repair (group B) were analyzed retrospectively. Videomanometry was conducted to investigate esophageal motor patterns. Esophageal acid exposure was evaluated with 24-hour esophageal pH monitoring. Data are expressed as medians and ranges. RESULTS: Contractions in the distal esophagus were conspicuously absent in 1 and 3 patients in groups A and B, respectively (P = .26). There were no significant differences in esophageal acid exposure (5.5% [0.7%-24.6%] vs 3.7% [0.3%-56.8%]; P = .71) or mean esophageal acid reflux time (0.5 minutes [0.1-1.4 minutes] vs 0.5 minutes [0.1-1.3 minutes]; P = .87) between the 2 groups. Fundoplication was conducted in 2 patients in each group (P = .60), all of whom had conspicuously absent distal esophageal contractions. Those contractions were preserved in the remaining patients with the exception of 1 group B patient. CONCLUSION: There are unlikely to be benefits from thoracoscopic repair of EA in terms of postoperative esophageal motor function. PMID- 20006011 TI - The measurement and designation of the pectus bar by computed tomography. AB - PURPOSE: The length of the bar used for the Nuss procedure is typically determined by measuring the distance between the 2 midaxillary lines and subtracting 2.5 cm. However, this may not be accurate for all patients. Measurements of the chest using computed tomography (CT) were developed for better determination of bar length. METHODS: Seventy-five patients underwent the Nuss procedure between 2005 and 2008. The length and curve of the pectus bar were determined using both the traditional method (TM) and CT. RESULTS: Twelve patients (16%) had length differences (LD) with the methods. The LD and non-LD patients were 18.8 +/- 2.4 and 11.3 +/- 0.6 years old, respectively (P = .005). The proportion of females in the groups was 58.3% (7/12) and 28.6% (18/63), respectively (P = .046). The TM gave a longer bar estimation in all but one patient with a high sternal angulation. Slight bar protrusion was noted in 3 of the 12 patients with LD using TM and 0 of the 9 patients using CT. CONCLUSIONS: Computed tomography measurement is a precise means for determining length, especially in older patients, females with developed breasts, or patients with high sternal angulation. It also allows for better curvature design, preventing multiple intraoperative adjustments. PMID- 20006012 TI - Analysis of changes to the anterior chest wall after the Nuss procedure-an objective measurement of pectus excavatum. AB - INTRODUCTION: We report a noninvasive method for recording the deviation of the anterior chest wall (ACW) in patients with pectus excavatum before and after the insertion and removal of a Nuss bar. PATIENTS AND METHOD: All patients undergoing the Nuss procedure at our institutions between June 2008 and February 2009 were enrolled in this study. A thermal plastic strip was used to cast the ACW at the skin level along the area corresponding to the tract of the inserted bar. The height of the ACW was defined as the perpendicular length from the xiphoid process to the transverse line drawn between the 2 midaxillary points. RESULTS: Bar insertion caused a significant increase in ACW height and width. Bar removal reduced the corrective effects in terms of a significant decrease in ACW height; at the same time, an increase in ACW width was noted. CONCLUSION: Feature extraction and analyses of permanent contour casts of the ACW help to understand immediate changes in the ACW after the Nuss procedure and may assist in the design of the bar at insertion. The Nuss bar in situ helped to maintain cosmetic benefits to the ACW. PMID- 20006013 TI - Connexin43 plays an important role in lung development. AB - OBJECTIVES: Connexin43 (Cx43) is one of the proteins associated with gap junction. Connexin43 knockout mice die after birth owing to hypoplastic lungs. The purpose of this study was to analyze the hypoplastic lung of Cx43 knockout mice to clarify the role of the Cx43 during lung development. METHODS: Adult hetero Cx43 mice were mated. Newborn mice were divided into the following groups: wild, hetero, and knockout. Total RNA was extracted from the right lung, and the left lung was fixed for immunohistochemical staining. The mRNA expression of surfactant protein C, aquaporin-5, and alpha-smooth muscle actin were analyzed by reverse transcriptase polymerase chain reaction. H&E and immunohistochemical staining for those markers were performed. RESULTS: The mRNA expression of aquaporin-5, surfactant protein C, and alpha-smooth muscle actin was significantly lower in knockout mice than that in the wild and hetero mice. H&E staining in the knockout mice showed narrow airspaces and thicker interalveolar septae. Immunohistochemical staining in all markers showed the formation of alveoli to be delayed in the knockout mice. CONCLUSION: Based on these findings, Cx43 is closely related to alveolar and vascular formation during lung development. PMID- 20006014 TI - Up-regulation of Wnt5a gene expression in the nitrofen-induced hypoplastic lung. AB - PURPOSE: The pathogenesis of pulmonary hypoplasia in nitrofen-induced congenital diaphragmatic hernia (CDH) still remains unclear. Wnt signaling pathways play a critical role in lung development. Whereas canonical Wnt signaling regulates branching morphogenesis during early lung development, the noncanonical Wnt5a controls late lung morphogenesis, including patterning of distal airway and vascular tubulogenesis (alveolarization). Overexpression of Wnt5a in transgenic mice and in the chick has been reported to result in severe pulmonary hypoplasia. We designed this study to test the hypothesis that the pulmonary Wnt5a gene expression is up-regulated in late stages of lung morphogenesis in CDH. METHODS: Pregnant rats were exposed to either olive oil or nitrofen on day 9 of gestation (D9). Fetal lungs were harvested on D15, D18, and D21 and divided into 3 groups: control; nitrofen without CDH, CDH(-); and nitrofen with CDH, CDH(+) (n = 8 at each time-point, respectively). Wnt5a pulmonary gene expression was analyzed by real-time reverse transcription polymerase chain reaction. Immunohistochemistry was performed to evaluate Wnt5a protein expression at each time-point. RESULTS: Pulmonary relative mRNA expression levels of Wnt5a were significantly increased in CDH(-) and CDH(+) at D18 (1.61 +/- 0.92 and 1.81 +/- 1.20, respectively) and D21 (2.40 +/- 0.74* and 2.65 +/- 0.35*, respectively) compared to controls at D18 and D21 (0.90 +/- 0.17* and 1.69 +/- 0.53**, respectively) (*P < .05, **P < .001 vs control ). Strong Wnt5a immunoreactivity was seen in the distal epithelium at D18 and D21 in nitrofen-induced hypoplastic lung compared to controls. CONCLUSION: Up-regulation of pulmonary Wnt5a gene expression in the late lung morphogenesis may interfere with patterning of alveolarization, causing pulmonary hypoplasia in the nitrofen-induced CDH. PMID- 20006015 TI - Management of choledochal cyst: 30 years of experience and results in a single center. AB - BACKGROUND: Choledochal cyst is usually diagnosed in childhood. Early treatment can prevent further complication. We report on our series of patients over the past 30 years. METHODS: A retrospective study was performed on all pediatric patients who presented with choledochal cyst from January 1978 to December 2008. The main outcome measures recorded were the clinical presentation, management, and long-term outcome of the patients. RESULTS: Eighty-three patients presented to us during the caption period with a mean age at diagnosis of 45 months (0 month to 16 years). Diagnoses were made antenatally in 15 patients. The most common symptoms were abdominal pain (n = 39) and jaundice (n = 35). Seventy-five patients had surgery, in which 72 patients had resection of the cyst and Roux-en Y hepaticojejunostomy. Ten were performed by laparoscopic means. We categorized the cysts based on the Todani classification. There was no mortality. No malignant change was documented. For those 4 who had Caroli disease, 2 underwent liver transplantation and 2 had hepatectomy. Overall early complication rate was 5.3% (4/75). CONCLUSIONS: Complete excision of cyst with Roux-en-Y hepaticojejunostomy is the treatment of choice, and the late result is good. Laparoscopic surgery is feasible. Long-term follow-up is necessary. There is no evidence to suggest that some type IV cysts are the result of disease progression from type I cysts. PMID- 20006016 TI - Excellent long-term outcome of hepaticojejunostomy for biliary atresia with a hilar cyst. AB - BACKGROUND: Biliary atresia (BA) with hilar cyst is an uncommon variant, which constitutes less than 10% of all types of BA, and the operative procedure for this type of BA remains controversial. METHODS: We have had 200 cases with BA from 1963 to 2008 in our institute and our branch hospitals, in which 12 cases (6%) were BA with a hilar cyst. The clinical records of all cases were evaluated retrospectively. RESULTS: Twelve BA patients with a hilar cyst included 2 boys and 10 girls. The diagnosis of BA was confirmed by intraoperative cholangiography (cloudy or treelike pattern). In all 12 cases, a hepaticojejunostomy was performed at a median age of 71.6 days (range, 24-136 days). The follow-up periods were 1.2 to 23.2 years. The current mean total bilirubin level was 0.8 mg/dL (range, 0.2-3.5 mg/dL), and the mean direct bilirubin level was 0.2 mg/dL (range, 0.0 to 0.8 mg/dL). Methylprednisolone or oral prednisolone was administered in 8 cases after operation, and 10 of 12 cases achieved a jaundice free state. The postoperative complications were cholangitis (n = 10), gastroesophageal varices (n = 7), splenomegaly (n = 3), ileus (n = 1), and pulmonary hypertension (n = 1). The overall survival rate with a native liver was 10 (83.3%) of 12 cases, of which 9 cases were jaundice-free and only 1 case showed recurrent jaundice. The other 2 cases underwent living-donor liver transplantation at age of 2 and 20 years, respectively, and they are currently doing well. CONCLUSIONS: Most cases of BA with a hilar cyst achieved excellent clinical outcome after a hepaticojejunostomy. PMID- 20006017 TI - Pediatric liver transplantation in Hong Kong-a domain with scarce deceased donors. AB - AIM: The study aimed to assess the outcome of live-donor liver transplantation for pediatric patients in a region with limited access to deceased donors. PATIENTS AND METHODS: From September 1993 to September 2008, 78 pediatric patients aged between 73 days and 17 years (mean, 40 months) received 83 liver transplants. Sixty-two were living-related liver transplantations (LRLTs), and 21 were deceased-donor liver transplantations (DDLTs). The mean follow-up period was 6.5 years. The prospectively collected data of these patients were analyzed retrospectively. RESULTS: The 1-, 2-, and 5-year survival rates of patients and grafts were 91%, 90%, 88% and 87%, 86%, 83%, respectively. The survival rates of LRLT patients and DDLT patients were 89%, 89%, 87%, and 90%, 86%, 86%, respectively (P = .58). The survival rates of patients aged 12 months or younger and patients older than 12 months were 95%, 92%, 90% and 90%, 90%, 87%, respectively (P = .65). One live donor developed temporary peroneal palsy, and another developed lung collapse (3%, 2/62). All live donors resumed their normal activities with no difficulty. CONCLUSION: With meticulous surgical techniques and postoperative care, it is justifiable to accept donated livers from voluntary live donors for transplantation to save pediatric patients in a place with scarce deceased donors. PMID- 20006018 TI - Direct vasodilative effect of FK506 on porcine mesenteric artery in small bowel transplantation. AB - BACKGROUND: Tacrolimus (FK506) is widely used as an immunosuppressive drug in small bowel transplantation. However, its precise effects on the vascular tone of the transplanted organ have not been studied. This study aimed to clarify the effects of FK506 on the porcine mesenteric artery. METHODS: The effects of FK506 on the changes in cytosolic Ca(2+) concentration ([Ca(2+)]i) and force using fura 2 fluorometry were investigated in mesenteric arterial strips of the porcine small intestine. The effects of FK506 on the activity of voltage-dependent Ca(2+) channels and receptor-operated Ca(2+) channels using high K(+) (118 mmol/L K(+)) depolarization and thromboxane A(2) analog (U46619) stimulation were also examined. RESULTS: FK506 inhibited the force development induced by 118 mmol/L K(+) depolarization and 1 micromol/L U46619 stimulation in a concentration dependent manner. The extent of inhibition of this contraction was greater than that of the K(+)-induced contraction, and its inhibitory potency was about 10 fold. FK506 (10 micromol/L) inhibited the increases in [Ca(2+)]i (24.9% +/- 7.4%) and the force development (52.0% +/- 5.6%) induced by 1 micromol/L U46619, respectively. CONCLUSIONS: FK506 induces arterial relaxation by decreasing [Ca(2+)]i. Pretreatment of a graft with FK506 may reduce the risk of vasospasm, ischemia-reperfusion injury, and thrombosis in small bowel transplantation. PMID- 20006019 TI - Inguinal hernias can be accurately diagnosed using the parent's digital photographs when the physical examination is nondiagnostic. AB - PURPOSE: Inguinal hernias in infants and children may not be evident during visits to the pediatric surgeon's office. Preoperative photographic confirmation of the hernia may be helpful and accurate. This study retrospectively assesses the preoperative examination, photographic evidence, operative findings, and complications of these patients. METHODS: For a 30-month period, children with a history of a possible inguinal bulge were assessed by clinical examination. If no bulge was detected, the parents were given the option of sending a photograph to the surgeon. Patients underwent surgery based solely upon the history and definitive image. RESULTS: For 30 months, 25 children were evaluated for inguinal pathology but had no hernia on clinical examination. Based solely upon the history and the digital image, 23 of these patients underwent surgery. All of the patients who were diagnosed with a hernia by history and imaging alone had an operatively confirmed hernia. The remaining 2 patients have not developed a hernia with continued observation. CONCLUSION: Photographic images are a reliable way to document the presence and laterality of inguinal hernias. Reliance upon digital transmission of images avoids the need for repeat office visits, saving considerable physician and parental time and expense. PMID- 20006020 TI - The antiandrogen flutamide perturbs inguinoscrotal testicular descent in the rat and suggests a link with mammary development. AB - AIM: Inadequate androgen activity is a likely cause of cryptorchidism in humans, affecting inguinoscrotal testicular descent. Flutamide, a nonsteroidal antiandrogen, produces cryptorchidism in rats. We aimed to determine the anatomical and histologic effects of flutamide. METHODS: Time-mated Sprague Dawley female rats were injected subcutaneously with flutamide (75 mg/kg in sunflower oil) on days 16 to 19 of pregnancy. Embryonic (E) and postnatal (P) male offspring were collected (E16, E19, P0, P2, P4, P8) in control and flutamide treated groups (n = 5-10). Samples were fixed in 4% paraformaldehyde. Five micrometer-thick sections were prepared for hematoxylin and eosin, trichrome and immunohistochemical stains (Desmin, TuJ1, Ki67). This identified muscle and neural cells and areas of cell proliferation. RESULTS: Postnatally, the gubernaculum in flutamide-treated rats had more mesenchyme and muscle than controls. Gubernacular eversion failed, and mammary tissue persisted around the gubernaculum in flutamide-treated rats. Flutamide had no effect on embryonic gubernacular anatomy and histology. CONCLUSIONS: Prenatal androgens altered postnatal gubernacular anatomy and histology in the postnatal period. Our findings indicate that the failure of gubernacular differentiation and migration may be because of the ongoing presence of mammary tissue in the region of the external inguinal ring. PMID- 20006021 TI - Mullerian-inhibiting substance deficiency in transgenic mice interferes with postnatal germ cell development: clues for understanding infertility in cryptorchidism. AB - AIM: Mullerian-inhibiting substance (MIS) may have a role in postnatal germ cell development, although this remains unproven. Elucidating the regulatory factors is crucial in finding new treatments for preventing infertility in cryptorchidism. We studied germ cell development in neonatal mice with MIS gene or receptor mutation to determine if germ cell development was affected. METHODS: Neonatal (5 MIS mutants, x1 MIS receptor mutant and 5 wild-type) and 10-day-old mice (x 7 MIS mutants, x1 MIS receptor mutant, 5 wild-type) were killed and prepared for hematoxylin-eosin and Masson trichrome histology of the testis. Testis diameter and tubule diameter were measured by Image-J, and germ cells were counted in 50 tubules/testis. RESULTS: Total testis and tubular diameters were greater in wild-type vs MIS mutants at days 0 and 10 (P < .01). Gonocytes were decreased in MIS mutants vs wild-type on day 0 (P = .019), and on day 10, the number of type A spermatogonia was slightly decreased (P = .05) and type B spermatogonia were significantly decreased (P < .01). Similar results were seen in the MIS receptor knockout. CONCLUSION: These results suggest that MIS has a previously unrecognized role in perinatal germ cell development that needs further investigation. Mullerian-inhibiting substance may be a possible future treatment for stimulating germ cell development in cryptorchidism. PMID- 20006022 TI - The expression of Cajal cells at the obstruction site of congenital pelviureteric junction obstruction and quantitative image analysis. AB - OBJECTIVE: The study aimed to analyze the expression and significance of interstitial cells of Cajal (ICC) at the obstruction site of congenital pelviureteric junction obstruction (PUJO). METHODS: Specimens of the strictured segment of the PUJ were obtained from 24 patients who were diagnosed to have PUJO (without aberrant vessels and adhesive band compression of ureteral junction) intraoperatively. In the control group, PUJ specimens were taken from 21 patients who had Wilms' tumor. Pelviureteric junction tissues were confirmed to be free of tumor invasion by histology. Immunohistochemistry with c-kit antibody was performed to detect the expression of ICC in specimens of the 2 groups. Quantitative analysis was made using image analysis technique and statistical analysis was carried out. RESULTS: Immunoreactivity to ICC was predominantly detected in the muscle layers of PUJ. The mean area of ICC expression in the PUJO group was 14.86 +/- 1.37 x 10(4)microm(2) , which was lower than that in the control group (16.80 +/- 1.68) x 10(4)microm(2) (P < .01). The mean density of ICC expression in the PUJO was 0.207 +/- 0.020, which was also lower than that in the control group (0.262 +/- 0.026) (P < .05). CONCLUSIONS: A reduction of the number of ICC may play an important role in the etiology and pathogenesis of PUJO. PMID- 20006023 TI - Neonatal small left colon syndrome in the offspring of diabetic mothers-an analysis of 105 children. AB - BACKGROUND AND AIM: Neonatal small left colon syndrome (NSLCS) is considered a rare cause of neonatal intestinal obstruction, with few documented cases in the English literature. Maternal diabetes has been reported in 40% to 50% of the published cases of NSLCS. Currently, the incidence of maternal diabetes is increasing, but there has been no study to ascertain the incidence and significance of NSLCS in this population. This review aims to assess the current significance of NSLCS and its association with maternal diabetes. MATERIALS AND METHOD: This was a retrospective review of 105 offspring of diabetic mothers who were admitted during the period 2004 to 2008 to our neonatal unit with special emphasis on associated NSLCS. RESULTS: There were 6 cases of intestinal obstruction in this group of 105 children. Five children, including a pair of twins, had classic features of NSLCS as demonstrated in the contrast enema with an abrupt transition at the splenic flexure and a narrow left colon. Conservative treatment was successful in all, negating the need for further invasive investigation. The sixth child had rectosigmoid Hirschprung disease. During the study period, there were no other cases of NSLCS in the nondiabetic population. CONCLUSION: Neonatal small left colon syndrome is the most common cause of intestinal obstruction in offspring of diabetic mothers. Neonatal small left colon syndrome can be confidently diagnosed in this population based on the classic clinical and radiologic findings. The incidence of NSLCS can be expected to increase as the incidence of maternal diabetes increases. PMID- 20006024 TI - Clinical features of Hirschsprung's disease associated with Down syndrome: a 30 year retrospective nationwide survey in Japan. AB - PURPOSE: To identify the clinical features in diagnosis and treatment of Hirschsprung's disease (HD) associated with Down syndrome (DS), the authors retrospectively analyzed data for patients with DS from the past 3 nationwide surveys in Japan. This survey was already approved by the Japanese Society of Pediatric Surgeons. METHODS: Patient data were collected in 3 phases-phase I (1978-1982), n = 47; phase II (1988-1992), n = 79; and phase III (1998-2002), n = 90. In total, data on 216 patients (5.6%) of 3852 were collected and analyzed. RESULTS: The incidence of DS in patients with HD was 2.9%, 7.1%, and 8.2% in phases I, II, and III, respectively, with a corresponding male/female ratio of 5:1, 2.4:1, and 5:1. The ratio of the extent of aganglionosis was nearly consistent across all phases. In phases I, II, and III, the incidence of total colonic aganglionosis was 2.1%, 0%, and 2.2%; and that of cardiovascular anomalies, 36.1%, 45.6%, and 55.6%; and that of preoperative enterocolitis, 31.0%, 26.6%, and 24.4%. The 2 most common surgical procedures were the Soave procedure, including transanal endorectal pull-through, and Duhamel procedure including Z-shaped anastomosis. The mortality rate decreased over time, from 26.1% in phase I to 11.4% in phase II and 7.8% in phase III. Almost all mortality cases were associated with cardiovascular anomalies: 54.5%, 62.5%, and 85.7% in phases I, II, and III, respectively. CONCLUSIONS: The incidence of HD with DS has increased over time. The number of male patients and cardiac anomalies has also increased in the last 10 years. Total colonic aganglionosis was rare. A marked decrease in the overall mortality rate was observed. PMID- 20006025 TI - An 18-year experience in total colonic aganglionosis: from staged operations to primary laparoscopic endorectal pull-through. AB - BACKGROUND: Traditionally, total colonic aganglionosis (TCA) was managed with enterostomy +/- pull-through. Since 1998, primary laparoscopic endorectal pull through (PLEP) was offered to patients with TCA in our unit. A retrospective study was conducted and early results reviewed. METHOD: Patient data were collected retrospectively. Before 1998, enterostomy followed by open Duhamel pull through was performed. From 1998, patients were assessed to receive staged operation or PLEP. RESULTS: From June 1990 to May 2007, 10 patients (6 males) were treated. All presented in the neonatal period. Seven patients, with transition zone within proximal 15 cm of terminal ileum, had pull-through performed. Three had staged pull-through, whereas 4 had PLEP. Two had extensive small bowel involvement with high output stoma pending reconstruction. One had total intestinal aganglionosis and died. The 3 patients with staged pull-through had normal bowel habit without soiling (mean follow-up, 13 years). For PLEP, all procedures were accomplished laparoscopically without intraoperative complication (mean operative time, 6 hours 50 minutes). The mean bowel motion is 5x per day, but most were too young to evaluate continence (mean follow-up, 3.5 years). CONCLUSION: Primary laparoscopic endorectal pull-through is feasible in TCA with limited small bowel involvement and has the advantage of a single-stage operation, avoiding a stoma and its related complications. PMID- 20006026 TI - A comparison of the effectiveness of the Soave and Martin procedures for the treatment of total colonic aganglionosis. AB - PURPOSE: Total colonic aganglionosis (TCA) is a relatively uncommon and severe condition managed by pediatric surgeons. Several procedures exist for the treatment of TCA. However, there is no current consensus on a superior operative procedure. The objective of this article is to evaluate the comparative effectiveness of the Soave procedure and the Martin procedure in the treatment of TCA with respect to preoperative data and postoperative outcome. METHODS: In the period from January 2001 to June 2008, we recruited 29 patients with TCA who were treated with the Martin procedure (14) or the Soave procedure (15). Relevant data were collected from the medical charts kept in the hospital library. The follow up study used a detailed questionnaire that was answered by patients either by telephone or directly in the clinic. In the clinic, data, including age, body weight before operation, intraoperative blood transfusion, anastomotic leakage, postoperative enterocolitis, durations of any postoperative fever, days in hospital, and a range of functional outcomes, were particularly recorded to enable a comparison of the effectiveness of the Soave procedure and the Martin procedure. RESULTS: A male predominance has been described for patients with TCA, with a male-to-female ratio of approximately 6.25:1. In the Soave group, time between ileostomy and the definitive procedure, body weight before the definitive procedure, and the operative age were less than those in the Martin group (P < .05). In patients who underwent the Martin procedure, the average duration of postoperative fever and days in hospital were significantly more than those in the Soave group (P < .05). In the Soave group, the incidence of intraoperative blood transfusion (46.7%), abdominal wound infection (6.67%), and anastomotic leakage (0) was significantly less (P < .05) than those in the Martin group (92.8%, 28.5%, and 14.3%, respectively). In the follow-up study, 7 patients in the Martin group had enterocolitis, and 4 of them had severe enterocolitis leading to multiple hospitalizations. In contrast, in the Soave group, only 2 patients had enterocolitis, and 1 was ameliorated after the therapy of anus dilation in the clinic. During the 4 weeks after the operation, all patients had an increased frequency of defecation, with soiling in the perianal area. At 3 months after the operation, there were significantly more patients with normal defecation in the Martin group compared to the Soave group (P < .05). Six months later, 11 (78.6%) patients in the Martin group and 8 (61.5%) patients in the Soave group had normal defecation and the difference was still significant (P < .05). CONCLUSION: Patients with TCA managed with the Soave procedure had fewer operative complications compared with those of patients who underwent the Martin procedure. However, the patients managed with the Soave procedure took longer to establish normal defecation. Although each procedure has advantages and disadvantages, the Soave procedure is promising for the treatment of TCA, especially with regard to postoperative complications. A long-term follow-up study is needed. PMID- 20006027 TI - Modified Soave procedure for the treatment of vascular malformations involving anorectum and sigmoid colon. AB - PURPOSE: The objective of this study is to introduce a modified Soave procedure for the treatment of vascular malformations involving the anorectum and sigmoid colon (VMARS) in children. METHODS: Cases of 12 children with VMARS between 2000 and 2008 were reviewed and analyzed. The confirmed diagnosis was established by barium enema, colonoscopy, computed tomography, and magnetic resonance imaging. All the patients underwent a modified Soave procedure. In 11 patients, Sarasola Klose hemorrhoidectomy was used for the distal part of endorectal dissection. RESULTS: The mean length of resected bowel was 22.5 cm, ranging from 17 to 28 cm. Histologically, the surgical specimens showed that the lesions were venous malformation. Postoperative recovery was uneventful. The patients had good continence with no rectal bleeding, but intermittent fecal soiling was noted in one case. CONCLUSIONS: VMARS forms a unique subset of patients with vascular malformations who have rectal bleeding. The combination of Soave procedure and Sarasola-Klose hemorrhoidectomy is a safe and effective procedure for VMARS. PMID- 20006028 TI - Neuronal immaturity in normoganglionic colon from cases of Hirschsprung disease, anorectal malformation, and idiopathic constipation. AB - AIM: Immaturity of neurons in normoganglionic colon in Hirschsprung disease (HD), anorectal malformation (AM), idiopathic constipation (IC), and normal controls (C) was assessed using polysialyated neural cell adhesion molecule. METHODS: Polysialyated neural cell adhesion molecule immunoreactivity in 3 sections of normoganglionic colon from HD (n = 48), AM (n = 25), IC (n = 36), and C (n = 18) were scored semiquantitatively according to age; 1 day to 11 months (G1), 1 to 4 years (G2), and 5 years and older (G3). RESULTS: Neurons in all specimens appeared mature irrespective of age on hematoxylin-eosin stain. Polysialyated neural cell adhesion molecule was positive (immaturity) in all specimens during G1 (1.34 in HD, 1.60 in AM, 0.89 in IC, and 1.59 in C) and decreased significantly with age in C (0.34* for G2, 0.25* for G3; *P < .01), decreased after 4 years old in IC (0.93 for G2, 0.10(#) for G3; (#)P < .05), decreased gradually in AM (1.10 for G2, 0.75( section sign) for G3; ( section sign)P < .05), but remained strongly positive in HD (1.34 for G1, 1.26 for G2, and 1.21 for G3; P = not significant), which after 4 years was significantly higher than C (P < .05). CONCLUSION: Postoperative colonic dysmotility may be because of persistence of immature neurons in HD and impaired maturation of neurons in AM and IC. PMID- 20006029 TI - Clinical value of pelvic 3-dimensional magnetic resonance image reconstruction in anorectal malformations. AB - OBJECTIVE: The study aimed to build a 3-dimensional (3D) reconstruction of pelvic magnetic resonance images and evaluate the clinical value in anorectal malformations (ARMs). METHODS: Magnetic resonance imaging (MRI) examinations were performed on a 1.5-T magnet. Sagittal, coronal, and transverse turbo spin-echo T1 weighted and fast spin-echo T2-weighted images of the pelvic region were obtained in 22 children. A 3D reconstruction was made on a computer and assisted by the 3D Doctor software (Trial Version, Able Software Corp). The level and type of ARM and the developmental state of the striated muscle complex (SMC) were analyzed with 3D reconstruction image. RESULTS: The 3D images of the pelvic were confirmed in 22 cases. Three-dimensional reconstructed images perfectly displayed the anatomical relationships of the SMC and the rectal atresia in these spaces. The 3D configuration of the SMC was different in each of the high- and low-type cases. The high-type malformation of SMCs differed particularly from the descriptions. CONCLUSIONS: Pelvic magnetic resonance 3D reconstructed images were able to show the dimensional anatomical relations of pelvis, bladder, urethra, rectum, and SMC. Both a 3D image and positional information with MRI offers the surgeon a simulated operative profile of the SMC superior to MRI slices alone, which will help in providing morphological data for image diagnosis and operation of the ARM. PMID- 20006030 TI - Unexpectedly deformed anal sphincter in low-type anorectal malformation. AB - PURPOSE: The sphincter muscles in anorectal anomalies, which do not appear in the surgical field, cannot be described adequately. Details of the anal sphincter in low-type anorectal malformations were investigated using 3-dimensional (3D) image analysis. PATIENTS AND METHODS: Thirty patients (10 males and 20 females) with low-type anorectal malformation were investigated with multidetector-row helical computed tomography. An image of the anorectal part was obtained with a slice thickness of 1 mm and a reconstruction pitch of 0.5 mm. A 3D reconstruction on a personal computer was made with a volume rendering method assisted by our own software (NewVES). RESULTS: Very thin vertical fibers (VFs) were observed behind the fistula in almost half of the low-type cases. They did not wrap the distal end of the fistula. The 3D images of these sphincters were different in each case. DISCUSSION: The fistula was dislocated forward from the deformed hypoplastic sphincter. We suggest that surgical mobilization of the anorectum into the center of the hypoplastic sphincter would be difficult using the cut back or Potts methods. CONCLUSION: The choice of surgical method should take into consideration the displaced and deformed hypoplastic anal sphincter. PMID- 20006031 TI - Comparison of clinical outcomes and anorectal manometry in patients with congenital anorectal malformations treated with posterior sagittal anorectoplasty and laparoscopically assisted anorectal pull through. AB - PURPOSE: The objective of this study is to analyze the clinical outcomes and anorectal manometry (AM) in infants with congenital high anorectal malformations treated with posterior sagittal anorectoplasty (PSARP) and laparoscopically assisted anorectal pull through (LAARP). MATERIALS AND METHODS: From August 2005 to December 2008, 23 patients with congenital high anorectal malformations were randomly distributed into PSARP and LAARP groups. All of them underwent LAARP (11 cases) or PSARP (12 cases) at 2 or 3 months old. Clinical outcomes and results of anorectal manometry were compared between patients at the age of 17.4 +/- 4.9 and 19.3 +/- 6.2 months (P = .4270), respectively. RESULTS: Kelly's clinical score for patients in LAARP and PSARP groups was 3.91 +/- 1.14 and 3.83 +/- 1.40 (P = .8827), respectively. Anal canal resting pressure and high-pressure zone length were 29.4 +/- 7.2 vs 23.4 +/- 6.5 mm Hg (P = .0479) and 14.9 +/- 3.0 vs 13.9 +/- 3.1 mm (P = .4414), respectively. Rectal anal inhibitory reflex was observed in 81.8% (9/11) and 83.3% (10/12) patients (P = 1.0000), respectively. The mean length of stay during the second hospitalization was 10.6 +/- 0.9 and 14.3 +/- 1.4 days (P < .0001), respectively. CONCLUSIONS: Although no significant difference can be noted in clinical scoring between both groups, the results of anorectal manometry indicate that LAARP can significantly improve anal canal resting pressure and reduce the length of stay. PMID- 20006032 TI - The effects of biofeedback training of pelvic floor muscles on fecal incontinence. AB - OBJECTIVE: The purpose of this study is to discuss the effect of pelvic floor muscle training on fecal incontinence. METHODS: A retrospective study was performed on patients who received pelvic floor muscle training from March 2002 to April 2007. There were 55 patients with fecal incontinence (male, 32 cases; female, 23 cases; mean age, 9.4 years old from 6 to 14), including 39 cases of anorectal malformation and 16 cases of Hirschsprung's disease. Pelvic floor muscle training was performed using biofeedback for 2 weeks in hospital, 2 times each day, and 30 minutes each time. The patients were then instructed to carry out self-training at home without the biofeedback device daily and received training evaluations in the hospital outpatient department monthly. All patients completed the training regimen and were followed up for 1 year. Anal manometry and clinical score were evaluated before and after training. RESULTS: Anal continence of 30 patients had satisfactory improvement, but not for the other 25 cases after training. The mean anal squeeze pressures of the group that had good results and the group that had poor results were 98.4 +/- 7.3 and 47.4 +/- 13.6 mm Hg, respectively, before training. There were 31 patients whose anal squeeze pressures were above 80 mm Hg, and 26 of these had satisfactory anal continence improvement, including all patients with Hirschsprung's disease. On the contrary, only 4 of 24 cases whose anal squeeze pressure was below 80 mm Hg acquired satisfactory anal continence improvement. CONCLUSIONS: Pelvic floor muscle training could achieve good results in some patients with fecal incontinence. Baseline measurements during anorectal manometry appear to provide good prediction of prognosis and effective management. PMID- 20006033 TI - Daily transabdominal electrical stimulation at home increased defecation in children with slow-transit constipation: a pilot study. AB - PURPOSE: Transcutaneous electrical stimulation (TES) (3 sessions/wk) over the abdomen stimulated bowel functions in a randomized controlled trial. This pilot study assessed whether daily TES at home with a safe, portable machine would be possible and more efficacious than trial results. METHODS: Eleven patients (6 male/5 female; mean age, 14 years; range, 12-18 years) with slow-transit constipation who relapsed or responded poorly in the trial were recruited (11 +/- 5 months later). An EPM-IF-4160 (Fuji Dynamics, Hong Kong) portable machine (sine waveform, 4 kHz carrier frequency, 80-160 Hz beat frequency, intensity <33 mA) delivering interferential current (2 electrodes over epigastrium + 2 over kidneys) was applied 1 hour daily at home. Continence diaries were kept for 1 month before and 2 months during treatment. RESULTS: All children completed more than 1 month of treatment after baseline recording. Defecation increased in 9 of 11 children, and soiling decreased in 4 of 11 children. There was a significant increase in total episodes of defecation per week (mean +/- SD, 2.5 +/- 2.1 vs 6.7 +/- 4.4; P = .008) and a nonsignificant decrease in soiling (3.8 +/- 1.6 vs 1.1 +/- 0.5 episodes/wk, P = .1). Daily stimulation does not affect abdominal pain. No adverse events occurred. CONCLUSIONS: Daily TES at home is safe and significantly improved bowel function in children who did not respond to 3 times per week of TES. Home TES may be a novel treatment of intractable slow transit constipation, avoiding hospital visits. PMID- 20006034 TI - 42nd Annual meeting of the Pacific Association of Pediatric Surgeons. PMID- 20006036 TI - Umbilical cyst containing ectopic gastric mucosa originating from an omphalomesenteric duct remnant. AB - An umbilical cyst originating from an omphalomesenteric duct remnant is extremely rare, and to the best of our knowledge, it has scarcely been reported in medical literature. We present ultrasonographic manifestations and computed tomographic findings of an umbilical cyst originating from an omphalomesenteric duct remnant in a 6-year-old girl. In cases of umbilical cyst, radiographic evaluation alone may not differentiate an omphalomesenteric duct remnant with heterotopic gastric mucosa from a urachal remnant, and surgical intervention is required. PMID- 20006035 TI - A 2-year-old boy with a stage III yolk sac tumor occurring in an intra-abdominal retained testis. AB - A major complication of retained testes is an occurrence of malignancy later in life. We, herein, report the case of a 2-year-old boy who presented with a huge yolk sac tumor with retroperitoneal lymph nodes metastasis that originated in a left intra-abdominal undescended testis. Computed tomography and magnetic resonance imaging showed a huge round tumor connecting to the left retroperitoneal lymph nodes with metastasis extending from the left pelvic region to the left renal hilum. The serum alpha-fetoprotein level was 36,528 ng/mL. The right abdominal tumor appeared to be a giant testis that had strangulated at the neck of the cord. The tumor had ruptured at the side of the left pelvic lymph node metastasis, and a yolk sac tumor was diagnosed from a histologic analysis of the resected specimens. Postoperative PEB chemotherapy was effective, and a complete surgical resection of the tumor was performed 3 months after the initial laparotomy. The pathologic findings showed fibrous tissue without any tumor cells. The patient has been doing well for 18 months after the radical operation. This case might be a coincidental association of a yolk sac tumor occurring in an undescended testis, which thus caused a delay in making an accurate diagnosis. PMID- 20006037 TI - Transoral closure of a perforation of the hypopharynx from blunt trauma. AB - Hypopharyngeal perforation secondary to blunt trauma is a rare injury. It can be managed operatively or expectantly without clear criteria for either approach. Here, we present a case of a 17-year-old adolescent boy who had a hypopharyngeal tear from direct blunt trauma to the anterior neck received during a high school football game. Physical examination demonstrated cervical crepitus, and neck radiograph revealed air in the retropharyngeal space. Rigid endoscopy diagnosed a 3-cm linear tear in the posterior hypopharynx. The tear was repaired transorally using laparoscopic instruments and visualized by a rigid endoscope, followed by anterior neck dissection and drain placement. Postoperatively, he was kept nil per os and received intravenous antibiotics. He was discharged home and returned to the football field the same fall. PMID- 20006038 TI - Individualized management of upper rectal atresia. AB - BACKGROUND/PURPOSE: Congenital colonic atresia (CA) or stenosis is an infrequent cause of low intestinal obstruction in the neonate. Atresias can occur at any level, and the management of CA is determined by the atretic site and by the presence or absence of associated anomalies. We report our experience dealing with upper rectal atresia during a 5-year period. METHODS: Between January 2004 and December 2008, 3 female newborns with upper rectal atresia with or without associated anomalies were treated. Modes of clinical presentation, methods of diagnosis, associated anomalies, alternative management techniques, and clinical outcome were retrospectively analyzed. RESULTS: All 3 patients had progressive abdominal distension, bilious vomiting, and failure to pass meconium. Contrast enema showed an atresia at the upper rectum in 2 patients. At laparotomy, case 1 was found to have type III atresia of the upper rectum. Resection of the dilated portion of the proximal colon with end sigmoid colostomy was accomplished in the neonatal period followed by a transanal mucosectomy with takedown of the colostomy and a pull-through procedure at age 3 months. Case 3 had multiple jejunoileal atresias and an upper rectal atresia. The initial management was multiple resections of atretic bowel and anastomoses and an end sigmoid colostomy. The secondary procedure was a takedown of the colostomy and transanal mucosectomy with a pull-through procedure. Case 2 had type I upper rectal atresia in association with imperforate anus complicated by colon perforation during performance of a distal colostogram leading to a complicated and protracted clinical course. All the patients are currently well with voluntary bowel movements, and one has occasional soiling with follow-up of 9 months to 3 years. CONCLUSIONS: Colon atresia, especially at the level of the upper rectum, is uncommon. Whether to proceed with an ostomy or to individualize the operative procedure according to the location of the atresia is still controversial. Transanal mucosectomy was a useful technique at the time of the definitive pull through for the treatment of upper rectal atresia. In cases of upper CA associated with imperforate anus, delay in diagnosis and potential complications may result if the diagnosis of upper rectal atresia is missed. PMID- 20006039 TI - Multiple-stage correction of caudal duplication syndrome: a case report. AB - Caudal duplication syndrome is a very rare congenital deformity. A 13-year-old boy was born with duplicated colon-rectum and anus, diphallus, hydronephrosis of left kidney with megaureter, double bladders and urethras, and vertebral abnormalities. Multiple-stage correction was performed to remove the duplicated colon and the mucosa of the duplicated rectum. A new colon was reconstructed. The left kidney and megaureter were excised. The septum in the bladders was removed to convert the double bladders into a single bladder. The double phalluses were fused into a single penis. After these staged procedures, the boy is now living a normal life. PMID- 20006040 TI - Autologous repair of isolated complete sternal cleft in an adolescent. AB - Isolated sternal clefts are rare congenital anomalies. Complete clefts are rarer still with only 23 cases being reported. We present an adolescent girl with a complete sternal cleft. The cleft was treated with autologous anterior perichondrial flaps and pectoralis advancement flaps. PMID- 20006041 TI - Left paraduodenal hernia causing small bowel obstruction in an adolescent patient. AB - Internal hernias are an uncommon cause of bowel obstruction, accounting for less than 1% of cases. Paraduodenal hernias, the most common type of internal hernias, are believed to be congenital in origin. They can be asymptomatic, cause chronic abdominal pain, or present with acute intestinal obstruction with strangulation and ischemia. We describe a case of left paraduodenal hernia found in a patient who presented with acute intestinal obstruction. PMID- 20006042 TI - Extrathoracic esophageal elongation (Kimura's technique): a feasible option for the treatment of patients with complex esophageal atresia. AB - AIM: The aim of this study was to evaluate the outcome of all patients who underwent an extrathoracic esophageal elongation (EEE) (Kimura's technique) and determine its role, among other surgical options, for the treatment of patients with complex esophageal atresia (EA) who have a previously created esophagostomy. METHODS: Between March 1997 and September 2008, we performed 20 EEEs. Twelve patients were males and 8 were females. The diagnoses were type C EA (n = 12), type A EA (n = 5), type B EA (n = 2), and type D EA (n = 1). Mean age at the initiation of the EEE was 10 months (range, 3-25 months). RESULTS: At the time of this report, 15 of the 20 patients have finished the treatment, 4 patients are still in the process of elongation, and one patient (premature, with a birth weight of 685 g) died before the final esophageal reconstruction. Of the 15 patients who finished the treatment, 12 (80%) completed it satisfactorily and 3 (20%) had to be prematurely interrupted. (In 2 patients, despite multiple attempts, the upper pouch could not be adequately elongated, and in one patient, an early perforation of the upper pouch precluded further elongations.) Of the 12 patients who completed the treatment satisfactorily, 10 (83%) are asymptomatic and exclusively on oral alimentation, whereas 2 (17%) have a pseudodiverticulum and esophageal dysmotility (requiring supplemental alimentation through a gastrostomy). Five of the 12 patients have gastroesophageal reflux (2 required a Nissen fundoplication and 3 are being treated medically). CONCLUSIONS: We believe that the EEE is a useful surgical option for a selected group of patients with complex long-gap EA who required a primary esophagostomy and also for patients with any type of EA who developed severe complications after a primary repair and required a secondary esophagostomy. With this technique, we avoided an esophageal replacement in 80% of cases, and given that the EEE does not invalidate a later esophageal replacement, we believe that the EEE is a feasible initial option for these patients. PMID- 20006043 TI - Modification of the Nuss procedure for pectus excavatum to prevent cardiac perforation. AB - PURPOSE: In a few patients, cardiac perforation and aortic injury have occurred during the Nuss procedure for pectus excavatum. The article details a modification of this procedure that enables the prevention of fatal complications. METHODS: Our subjects were 22 males and 13 females with pectus excavatum who were aged 8.2 +/- 3.7 years. Their Haller's computed tomography index was 5.2 +/- 1.5. An introducer is inserted into the pleura between the sternum and thymus instead of the thoracic depression under right thoracoscopic guidance. After the introducer reaches the internal cranial position of the left nipple, the thoracoscope is shifted to the left thoracic cavity. The introducer is subsequently guided to the left highest intercostal space under left thoracoscopic guidance. RESULTS: A single bar was inserted in 34 patients; 2 bars were required in 1 patient. The operating time was 95 +/- 27 minutes and blood loss was 11 +/- 6 g. Cardiac perforation did not occur in any patient. CONCLUSIONS: Our modified technique has certain advantages: (1) the introducer does not rub against the pericardium and heart; (2) the tip of the introducer can be observed at all times with a thoracoscope; (3) the anterior mediastinum between the left and right thoracic cavities is very narrow at the cranial level; (4) the introducer can be accurately directed to the left highest intercostal space; and (5) hemostasis and no injury of the bilateral thoracic organs can be confirmed. PMID- 20006044 TI - Heterotopic pancreas-an unusual cause of melena in a pediatric patient. PMID- 20006045 TI - A rare anatomical variation of type B esophageal atresia. PMID- 20006046 TI - Zero-total event trials and incomplete pyloromyotomy. PMID- 20006048 TI - Does cellular heterogeneity influence neuroblastoma cell line proliferation and invasiveness with granulocyte colony-stimulating factor? PMID- 20006050 TI - References and other cited material. PMID- 20006052 TI - A review of chromatographic methods for determination of synthetic food dyes. AB - The purpose of this work was to present a chromatographic methods to analyse synthetic food dyes. The following techniques has been described: thin-layer liquid chromatography (TLC), high performance thin-layer chromatography (HPTLC), traditional column chromatography, high performance liquid chromatography (HPLC), include: ion-pair chromatography (HPLC IP), reversed phase chromatography (RP HPLC) and high performance ion chromatography. PMID- 20006053 TI - Application of chemometric methods to environmental analysis of organic pollutants: A review. AB - Organic pollutants include a very wide variety of chemical compounds with different structures, properties, functions and origins, which may produce diverse damages to the ecosystem and the human beings. This review presents the recent progress on the use of chemometrics to evaluate the occurrence of these substances in the environment. The main topics addressed are: (a) the problems related to the interpretation of the analytical measurements used in the determination of organic pollutants (quantitative analytical determinations section), (b) the profiling of the related environmental pollution sources through their compositional, geographical and temporal distribution patterns (environmental exploratory studies section) and (c) the prediction of the toxicological activity of these substances through models based on the use of structural or physical/chemical descriptors (toxicity studies section). Each section includes selected works related to pesticides, polycyclic aromatic hydrocarbons and other organic pollutants. PMID- 20006051 TI - Treatment of nonarteritic anterior ischemic optic neuropathy. AB - Nonarteritic anterior ischemic optic neuropathy (NAION) is the most common clinical presentation of acute ischemic damage to the optic nerve. Most treatments proposed for NAION are empirical and include a wide range of agents presumed to act on thrombosis, on the blood vessels, or on the disk edema itself. Others are presumed to have a neuroprotective effect. Although there have been multiple therapies attempted, most have not been adequately studied, and animal models of NAION have only recently emerged. The Ischemic Optic Neuropathy Decompression Trial, the only class I large multicenter prospective treatment trial for nonarteritic anterior ischemic optic neuropathy, found no benefit from surgical intervention. One recent large, nonrandomized controlled study suggested that oral steroids might be helpful for acute NAION. Others recently proposed interventions are intravitreal injections of steroids or anti-vascular endothelial growth factor (anti-VEGF) agents. There are no class I studies showing benefit from either medical or surgical treatments. Most of the literature on the treatment of NAION consists of retrospective or prospective case series and anecdotal case reports. Similarly, therapies aimed at secondary prevention of fellow eye involvement in NAION remain of unproven benefit. PMID- 20006054 TI - The electrochemical determination of phenolic derivates using multiple pulsed amperometry with graphite based electrodes. AB - The electrochemical determinations of 4-chlorophenol (4-CP) and 4-nitrophenol (4 NP) by chronoamperometry (CA) and multiple pulsed amperometry (MPA) using expanded graphite-epoxy composite (EG-Epoxy) and rotating spectral graphite disc (SG) electrodes are reported. The electrochemical behaviours of both electrodes in the presence of organics informed about oxidation peak potential and the electrode fouling with organics concentration increasing. Setting up the oxidation peak potential as detection potential, only SG gave good electroanalytical performance using CA. However, by MPA applying both electrodes exhibited the capability to assess electrochemically and quantitatively the pollutants from aqueous solutions. UV spectrometric method detecting 4-CP and 4 NP at lambda=280nm and lambda=398nm wavelength, respectively was used for validation and parallel determinations. PMID- 20006055 TI - Modelling of acid-base equilibria in binary-solvent systems: A comparative study. AB - A new approach to modelling of some binary (hydro-organic, HL/H(2)O+B) acid-base systems with organic co-solvent B fully miscible with water, is suggested and applied for the determination of acidity parameters pK for some weak acids HL. The models are designed to get the pK=pK(x) relationships (x-mole fraction of B in H(2)O+B) from results of pH titrations made in such systems and for the determination of pK(B)=pK(1) for HL in pure B. The Redlich-Kister equation, together with its asymmetric extensions, and the Legendre functions with orthogonal polynomials, appeared to be suitable for such purposes. PMID- 20006056 TI - Developing automated analytical methods for scientific environments using LabVIEW. AB - The development of new analytical techniques often requires the building of specially designed devices, each requiring its own dedicated control software. Especially in the research and development phase, LabVIEW has proven to be one highly useful tool for developing this software. Yet, it is still common practice to develop individual solutions for different instruments. In contrast to this, we present here a single LabVIEW-based program that can be directly applied to various analytical tasks without having to change the program code. Driven by a set of simple script commands, it can control a whole range of instruments, from valves and pumps to full-scale spectrometers. Fluid sample (pre-)treatment and separation procedures can thus be flexibly coupled to a wide range of analytical detection methods. Here, the capabilities of the program have been demonstrated by using it for the control of both a sequential injection analysis - capillary electrophoresis (SIA-CE) system with UV detection, and an analytical setup for studying the inhibition of enzymatic reactions using a SIA system with FTIR detection. PMID- 20006057 TI - An osmotic micro-pump integrated on a microfluidic chip for perfusion cell culture. AB - A novel microfluidic chip integrating an osmosis-based micro-pump was developed and used for perfusion cell culture. The micro-pump includes two sealed chambers, i.e., the inner osmotic reagent chamber and the outer water chamber, sandwiching a semi-permeable membrane. The water in the outer chamber was forced to flow through the membrane into the inner chamber via osmosis, facilitating continuous flow of fluidic zone in the channel. An average flow rate of 0.33microLmin(-1) was obtained within 50h along with a precision of 4.3% RSD (n=51) by using a 100mgmL(-1) polyvinylpyrrolidone (PVP) solution as the osmotic driving reagent and a flow passage area of 0.98cm(2) of the semi-permeable membrane. The power free micro-pump has been demonstrated to be pulse-free offering stable flow rates during long-term operation. The present microfluidic chip has been successfully applied for the perfusion culture of human colorectal carcinoma cell by continuously refreshing the culture medium with the osmotic micro-pump. In addition, in situ cell immunostaining was also performed on the microchip by driving all the reagent zones with the integrated micro-pump. PMID- 20006058 TI - Poly(malachite green) at nafion doped multi-walled carbon nanotube composite film for simple aliphatic alcohols sensor. AB - Conductive composite film which contains nafion (NF) doped multi-walled carbon nanotubes (MWCNTs) along with the incorporation of poly(malachite green) (PMG) has been synthesized on glassy carbon electrode (GCE), gold and indium tin oxide (ITO) electrodes by potentiostatic methods. The presence of MWCNTs in the composite film (MWCNTs-NF-PMG) enhances surface coverage concentration (Gamma) of PMG to approximately 396%, and increases the electron transfer rate constant (k(s)) to approximately 305%. Similarly, electrochemical quartz crystal microbalance study reveals the enhancement in the deposition of PMG at MWCNTs-NF film. The surface morphology of the composite film deposited on ITO electrode has been studied using scanning electron microscopy (SEM) and scanning tunneling microscopy (STM). These two techniques reveal that the PMG incorporated on MWCNTs NF film. The MWCNTs-NF-PMG composite film also exhibits promising enhanced electrocatalytic activity towards the simple aliphatic alcohols such as methanol, ethanol and propanol. The electroanalytical responses of analytes at NF-PMG and MWCNTs-NF-PMG films were measured using both cyclic voltammetry (CV) and differential pulse voltammetry (DPV). From electroanalytical studies, well defined voltammetric peaks have been obtained at MWCNTs-NF-PMG composite film for methanol, ethanol and propanol at Epa=609, 614 and 602mV respectively. The sensitivity of MWCNTs-NF-PMG composite film towards methanol, ethanol and propanol in CV technique are 0.59, 0.36 and 0.92microAmM(-1)cm(-2) respectively, which are higher than NF-PMG film. Further, the sensitivity values obtained using DPV are higher than the values obtained using CV technique. PMID- 20006059 TI - Comparison between ordinary least squares regression and weighted least squares regression in the calibration of metals present in human milk determined by ICP OES. AB - In this study we compared the use of ordinary least squares and weighted least squares in the calibration of the method for analyzing essential and toxic metals present in human milk by ICP-OES, in order to avoid systematic errors in the measurements used. Human milk samples were provided by maternity clinic Odete Valadares and digested by means of a high-performance microwave (MW) oven. Evaluation of plasma short and long-term stability was made using a solution of digested milk (1:50) with 2.0mgL(-1) Mg in HNO(3) 2% (v/v). The detection power resulted to be at or below the mugL(-1) level, whilst the precision expressed as relative standard deviation R.S.D. was almost always equal to or better than 3.3%. Certified reference material Infant Formula (NIST SRM 1846) was used to assess the accuracy of the proposed method, which proved to be accurate and precise. Recovery rates were in the range of 83-117%. Aqueous calibration was carried out for each element under study. PMID- 20006060 TI - A liquid chromatography-electrospray ionization-tandem mass spectrometry study of ethanolamines in high salinity industrial wastewaters. AB - The detection and quantitation of four ethanolamines, tris(2-hydroxyethyl)amine (triethanolamine, TEA), N,N-bis(2-hydroxyethyl)methylamine (methyldiethanolamine, MDEA), N-(2-aminoethyl)ethanolamine (AEA), and N,N-diethylethanolamine (DEA), were achieved in wastewaters from two aerobic activated sludge bioreactors located in an industrial wastewater treatment plant. The streams had salt concentrations of approximately 3% and 7% by weight in Reactor 1 and Reactor 2, respectively. The use of liquid chromatography-electrospray ionization-tandem mass spectrometry avoided the need for some sample preparation steps such as extraction, concentration, and derivatization. Ion suppression in the electrospray, attributable to the presence of sodium clusters, was attenuated by a 10-fold dilution of the wastewaters with acetonitrile. A matrix-matched calibration model averted other potential interferences. For the compounds analyzed in selected reaction monitoring mode (TEA, MDEA, and DEA), the calibration curves presented linearity in a range of 10-1000microg/L with corresponding detection limits ranging from 2 to 11microg/L, depending upon the specific analyte and aqueous matrix. AEA was calibrated in selected ion monitoring mode (100-1000microg/L), with corresponding detection limits in the two wastewaters of 74.6 and 85.3microg/L, respectively. Overall good precision (<10%) and accuracy (97-110%) were achieved for both matrices, which fell within laboratory reproducibility. Finally, the amines were introduced into six mixed liquor samples from both reactors and quantified following the reported protocol. Again, recoveries were close to 100% with a relative standard deviation of less than 10% in all cases. PMID- 20006061 TI - Supercritical fluid extraction of polybrominated diphenyl ethers (PBDEs) from house dust with supercritical 1,1,1,2-tetrafluoroethane (R134a). AB - The extraction of polybrominated diphenyl ethers (PBDEs) from SRM 2585 (Organic Contaminants in House Dust) was investigated using supercritical fluid R134a as an extraction solvent. Three methods of dust extraction were studied: (1) extraction of dry dust, (2) extraction of dry dust dispersed on Ottawa sand and (3) extraction of dust wet with dichloromethane. For each of the three sample preparation methods, extracts at three temperatures (110, 150, and 200 degrees C) above the critical temperature of R134a were performed. Eight PBDE congeners (BDE 28, -47, -99, 100, -153, -154, -183, and -209) in the SFE extracts were analyzed by liquid chromatography negative-ion atmospheric pressure photoionization tandem mass spectrometry (LC/NI-APPI/MS/MS). The optimum extraction of PBDEs from house dust using supercritical R134a is obtained when the dust is pre-wet with dichloromethane prior to extraction to swell the dust. For all sample preparation methods, higher temperatures afforded higher percent recoveries of the eight PBDE congeners. Only a combination of high-temperature (200 degrees C) and pre-wetting the dust with dichloromethane produced high recovery of the environmentally important, fully brominated PBDE congener, BDE-209. PMID- 20006062 TI - Assessment uncertainty associated to the analysis of organic compounds linked to particulate matter of atmospheric aerosols. AB - Current work has evaluated uncertainty associated to quantification of several organic compounds present in particulate matter of atmospheric aerosols, setting out the stages of analytical procedure that contribute most to the global uncertainty. Several sources of uncertainty have been identified, which were clustered into five main contributions: sampling, extraction, clean-up, derivatization and analysis. A discussion of the main contributions to the overall uncertainty is reported, allowing authors to locate the largest ones and plan future improvements. Combined uncertainties ranged between 10-18% (alkanes), 12-16% (PAHs), 10-18% (alcohols) and 9-21% (acids). The analytical procedure was validated by analysing a standard reference material (SRM1649a, urban dust). Also, the proposed method was applied to the analysis of four samples of particulate matter. PMID- 20006063 TI - Preparation of a new solid state fluoride ion selective electrode and application. AB - A new solid state fluoride ion selective electrode composed of 70% Ag(2)S, 10% Cu(2)S and 20% CaF(2) has been developed. An analytically useful potential change occurred, from 1x10(-6) to 1x10(-1)M fluoride ion. The slope of the linear portion (1x10(-1)-1x10(-5)M) was about 26+/-2mV/10-fold concentration changes in fluoride. It was found that pH change between 1 and 8 had no effect on the potential of the electrode. There was no interference of most common cations such as K(+,) Na(+), Ca(2+) and Mg(2+) and anions such as Cl(-), NO(3)(-), SO(4)(2-) and PO(4)(3-). The lifetime of the electrode was more than 2 years, when used at least 4-5 times a day, and the response time was about 60s. The measurements were made at constant ionic strength (0.1M NaNO(3)) and at room temperature. This electrode has been used for the determination of fluoride ion in Ankara city tap water and in bottled spring water using standard addition method. The validation of the electrode has been made with a commercial fluoride ion selective electrode (Orion) and high consistency was obtained. PMID- 20006064 TI - Determination of cadmium in alcohol fuel using Moringa oleifera seeds as a biosorbent in an on-line system coupled to FAAS. AB - In this study a new method for determination of cadmium in alcohol fuel using Moringa oleifera seeds as a biosorbent in an on-line preconcentration system coupled to flame atomic absorption spectrometry (FAAS) was developed. Flow and chemical variables of the proposed system were optimized through multivariate designs. The limit of detection for cadmium was 5.50microg L(-1) and the precision was below 2.3% (35.0microg L(-1), n=9). The analytical curve was linear from 5 to 150microg L(-1), with a correlation coefficient of 0.9993. The developed method was successfully applied to spiked alcohol fuel, and accuracy was assessed through recovery tests, with recovery ranging from 97.50 to 100%. PMID- 20006065 TI - A green and sensitive method to determine phenols in water and wastewater samples using an aqueous two-phase system. AB - A greener and more sensitive spectrophotometric procedure has been developed for the determination of phenol and o-cresol that exploits an aqueous two-phase system (ATPS) using a liquid-liquid extraction technique. An ATPS is formed mostly by water and does not require organic solvent. Other ATPS components used in this study were the polymer, polyethylene oxide, and some salts (i.e., Li(2)SO(4), Na(2)SO(4) or K(2)HPO(4)+KOH). The method is based on the reaction between phenol, sodium nitroprusside (NPS) and hydroxylamine hydrochloride (HL) in an alkaline medium (pH 12.0), producing the complex anion [Fe(2)(CN)(10)](10-) that spontaneously concentrates in the top phase of the system. The linear range was 1.50-500microgkg(-1) (R>or=0.9997; n=8) with coefficients of variation equal to 0.38% for phenol and 0.30% for o-cresol (n=5). The method yielded limits of detection (LODs) of 1.27 and 1.88microgkg(-1) and limits of quantification (LOQs) of 4.22 and 6.28microgkg(-1) for phenol and o-cresol, respectively. Recoveries between 95.7% and 107% were obtained for the determination of phenol in natural water and wastewater samples. In addition, excellent agreement was observed between this new ATPS method and the standard 4-aminoantipyrine (4-AAP) method. PMID- 20006066 TI - A novel molecularly imprinted polymer thin film as biosensor for uric acid. AB - A novel amine-imide type conducting polymer, denoted as poly(PD-BCD), was molecularly imprinted on an indium-tin oxide (ITO) glass, with uric acid (UA) as the template and without any functional monomer. Intending to improve the imprinting efficiency, the polymer content was varied from 0.3 to 0.9wt% during the preparation of the molecularly imprinted polymer (MIP), thereby varying the thickness of the polymer film; the content of UA as the template was maintained to be the same for all the films. The sensitivities of the thus prepared MIP electrodes were calculated to be more than 3-fold, compared to those of the corresponding non-MIP (NMIP) electrodes, which were obtained through the same method, however, without adding UA during their preparation. A polymer content of 0.6wt% rendered the best performing MIP electrode, as judged by the imprinting efficiency and sensitivity of the electrode for UA. A linear relationship between steady-state currents and UA concentrations from 0 to 1.125mM was obtained for both types of the sensors. The sensitivities of the MIP and the NMIP electrodes made with 0.6wt% of polymer were calculated to be 24.72 and 6.63microAmM(-1)cm( 2), respectively. The limit of detection (LOD) for this MIP was found to be 0.3microM at a signal to noise ratio (S/N) of 3. This MIP electrode was used as a biosensor for the detection of UA in the presence of ascorbic acid (AA) in a sample containing these species in the same concentrations as those in a human serum. The selectivity of MIP electrode is higher than that of NMIP electrode, and the values are 28.76 and 8.85, respectively. The results are substantiated by using cyclic voltammetry (CV), linear sweep voltammetry, amperometry, and scanning electron microscopy. PMID- 20006067 TI - Determination of non-steroidal anti-inflammatory drugs in wastewaters by magnetic matrix solid phase dispersion-HPLC. AB - A series of supports functionalized with different alkyl chains and covered with magnetite were synthesized, characterized and applied in the sample pre concentration of four non-steroidal anti-inflammatory drugs (acetaminophen, naproxen, diclofenac and ibuprofen) contained in wastewater samples. The general methodology involved magnetic solid phase dispersion followed by the analysis by high-performance liquid chromatography with ultraviolet detection. The magnetic supports were initially dispersed in the samples with the aid of Triton X-100, then supports were magnetically isolated and the analytes were eluted with methanol. Finally the extract was injected into the HPLC system. The highest recovery percentage (>90%) was obtained with the support containing octyl chains (C8) at pH 3. The lowest limits of detection achieved ranged within 1-2microgL( 1) with repeatability (expressed as RSD) below 5% in all cases. The method was applied in the analysis of wastewater samples. PMID- 20006068 TI - Methylmercury and inorganic mercury determination in blood by using liquid chromatography with inductively coupled plasma mass spectrometry and a fast sample preparation procedure. AB - Despite the necessity to differentiate chemical species of mercury in clinical specimens, there are a limited number of methods for this purpose. Then, this paper describes a simple method for the determination of methylmercury and inorganic mercury in blood by using liquid chromatography with inductively coupled mass spectrometry (LC-ICP-MS) and a fast sample preparation procedure. Prior to analysis, blood (250microL) is accurately weighed into 15-mL conical tubes. Then, an extractant solution containing mercaptoethanol, l-cysteine and HCl was added to the samples following sonication for 15min. Quantitative mercury extraction was achieved with the proposed procedure. Separation of mercury species was accomplished in less than 5min on a C18 reverse-phase column with a mobile phase containing 0.05% (v/v) mercaptoethanol, 0.4% (m/v) l-cysteine, 0.06molL(-1) ammonium acetate and 5% (v/v) methanol. The method detection limits were found to be 0.25microgL(-1) and 0.1microgL(-1) for inorganic mercury and methylmercury, respectively. Method accuracy is traceable to Standard Reference Material (SRM) 966 Toxic Metals in Bovine Blood from the National Institute of Standards and Technology (NIST). The proposed method was also applied to the speciation of mercury in blood samples collected from fish-eating communities and from rats exposed to thimerosal. With the proposed method there is a considerable reduction of the time of sample preparation prior to speciation of Hg by LC-ICP MS. Finally, after the application of the proposed method, we demonstrated an interesting in vivo ethylmercury conversion to inorganic mercury. PMID- 20006069 TI - Multi-wall carbon nanotubes (MWCNTs)-doped polypyrrole DNA biosensor for label free detection of genetically modified organisms by QCM and EIS. AB - In this paper, we describe DNA electrochemical detection for genetically modified organism (GMO) based on multi-wall carbon nanotubes (MWCNTs)-doped polypyrrole (PPy). DNA hybridization is studied by quartz crystal microbalance (QCM) and electrochemical impedance spectroscopy (EIS). An increase in DNA complementary target concentration results in a decrease in the faradic charge transfer resistance (R(ct)) and signifying "signal-on" behavior of MWCNTs-PPy-DNA system. QCM and EIS data indicated that the electroanalytical MWCNTs-PPy films were highly sensitive (as low as 4pM of target can be detected with QCM technique). In principle, this system can be suitable not only for DNA but also for protein biosensor construction. PMID- 20006070 TI - Solid-phase extraction of organic compounds in atmospheric aerosol particles collected with the particle-into-liquid sampler and analysis by liquid chromatography-mass spectrometry. AB - Atmospheric aerosol particles, collected with the particle-into-liquid sampler at SMEARII station in Finland in mid-August 2007, were analysed for biogenic acids. The sample pretreatment method, comprising solid-phase extraction with anion exchange and hydrophilic-lipophilic balance materials, was optimized. Extraction efficiencies of solid-phase extraction from 10 and 20ml samples were about 100%, with average relative standard deviation of 8.9%, in concentration range from 12.5 to 50ng/ml of the acid. Extraction of aldehydes was less successful, with efficiencies from 69 to 163% and average 10% deviation. Pretreated samples were analysed by reversed phase high performance liquid chromatography with ion trap mass spectrometric detection. Limits of detection achieved for organic acids with the analytical procedure developed ranged from 9 to 27microg/l of extracted sample, while limits of quantitation were from 31 to 90microg/l. Oxidation with ozone was used for the preparation of the acid of beta-caryophyllene (beta caryophyllinic acid), which was also studied in aerosol samples. MS(2) experiments were used to confirm the identification of trans-pinic, trans-pinonic and beta-caryophyllinic acids. Azelaic, hexadecanoic, cis-pinonic, and cis- and trans-pinic acids were quantitated in the samples with use of authentic standards, while the concentrations of trans-pinonic and beta-caryophyllinic acids were determined with cis-pinonic acid as surrogate. Also, the contribution of beta-caryophyllene oxidation products to aerosol organic carbon was evaluated. Aldehydes could not be analysed in real samples due to the insufficient extraction. The particle-into-liquid sampler proved to be suitable for the collection of aerosol particles for the elucidation of daily and diurnal variation of selected species. The optimized sample pretreatment, together with the analysis method, offer a promising approach for the study of aerosol chemical composition, where artifact formation is minimal and time resolution is good. PMID- 20006071 TI - Task-specific ionic liquid trioctylmethylammonium salicylate as extraction solvent for transition metal ions. AB - A quaternary ammonium-based room temperature ionic liquid trioctylmethylammonium salicylate (TOMAS) has been studied as an extractant of transition metal ions (Fe(3+), Cu(2+), Ni(2+), Mn(2+)) in aqueous solutions. The effect of pH value on the recovery of metal ions has been investigated. The mechanism of extraction into the ionic liquid has been proposed. The possibility of stripping voltammetric determination of transition metals in aqueous solutions using TOMAS modified electrodes has been demonstrated. PMID- 20006072 TI - Extraction of biogenic amines using sorbent materials containing immobilized crown ethers. AB - Three sorbent materials (A18C6-MS, DA18C6-MS and AB18C6-MS) based on the crown ether ligands, 1-aza-18-crown-6, 1,4,10,13-tetraoxa-7,16-diazacyclo octadecane and 4'-aminobenzo-18-crown-6, respectively, were prepared by the chemical immobilization of the ligand onto mesoporous silica support. The sorbents were characterized by FT-IR, scanning electron microscopy-energy dispersive X-ray microanalysis, elemental analysis and nitrogen adsorption-desorption test. The applicability of the sorbents for the extraction of biogenic amines by the batch sorption method was extensively studied and evaluated as a function of pH, biogenic amines concentration, contact time and reusability. Under the optimized conditions, all the sorbents exhibited highest selectivity toward spermidine (SPD) compared to other biogenic amines (tryptamine, putrescine, histamine and tyramine). Among the sorbents, AB18C6-MS offer the highest capacity and best selectivity towards SPD in the presence of other biogenic amines. The AB18C6-MS sorbent can be repeatedly used three times as there was no significant degradation in the extraction of the biogenic amines (%E>85). The optimized procedure was successfully applied for the separation of SPD in food samples prior to the reversed-phase high performance liquid chromatography separation. PMID- 20006073 TI - Simultaneous spectrophotometric determination of trace amounts of uranium, thorium, and zirconium using the partial least squares method after their preconcentration by alpha-benzoin oxime modified Amberlite XAD-2000 resin. AB - A new solid phase extraction method for separation and preconcentration of trace amounts of uranium, thorium, and zirconium in water samples is proposed. The procedure is based on the adsorption of U(VI), Th(IV) and Zr(IV) ions on a column of Amberlite XAD-2000 resin loaded with alpha-benzoin oxime prior to their simultaneous spectrophotometric determination with Arsenazo III using orthogonal signal correction partial least squares method. The enrichment factor for preconcentration of uranium, thorium, and zirconium was found to be 100. The detection limits for U(VI), Th(IV) and Zr(IV) were 0.50, 0.54, and 0.48microgL( 1), respectively. The precision of the method, evaluated as the relative standard deviation obtained by analyzing a series of 10 replicates, was below 4% for all elements. The practical applicability of the developed sorbent was examined using synthetic seawater, natural waters and ceramic samples. PMID- 20006074 TI - Langmuir-Blodgett film of p-tert-butylthiacalix[4]arene modified glassy carbon electrode as voltammetric sensor for the determination of Hg(II). AB - The pi-A isotherms and UV-vis spectra of the transferred films suggested that the monolayer of p-tert-butylthiacalix[4]arene can coordinate with Hg(2+) at the air water surface. From these observations, a glassy carbon electrode coated with Langmuir-Blodgett film of p-tert-butylthiacalix[4] arene as a new voltammetric sensor is designed for the determination of trace amounts of Hg(2+). Compared with bare glassy carbon electrode and modified glassy carbon electrode using direct coating method, the Langmuir-Blodgett film-modified electrode can greatly improve the measuring sensitivity of Hg(2+). Under the selected conditions, the Langmuir-Blodgett film-modified electrode in 0.1molL(-1) H(2)SO(4)+0.01molL(-1) KCl solution shows a linear voltammetric response for Hg(2+) in the range of 5.0x10(-10) to 1.5x10(-7)molL(-1), with a detection limit of 2.0x10(-10)molL(-1). The proposed method was also applied to determine Hg(2+) in water samples (tap, lake and river water). In addition, the fabricated electrode exhibited a distinct advantage of simple preparation, non-toxicity, good reproducibility and good stability. PMID- 20006075 TI - Silica gel modified with 1-(2-aminoethyl)-3-phenylurea for selective solid-phase extraction and preconcentration of Sc(III) from environmental samples. AB - A new method that utilizes 1-(2-aminoethyl)-3-phenylurea-modified silica gel as a solid-phase extractant has been developed for preconcentration of trace Sc(III) prior to the measurement by inductively coupled plasma atomic emission spectrometry (ICP-AES). Experimental conditions for effective adsorption of trace level of Sc(III) were optimized using batch and column procedures in detail. The optimum pH value for the separation of Sc(III) on the new sorbent was 4 and complete elution of Sc(III) from the sorbent surface was carried out using 1.0mL of 0.1molL(-1) HCl. Common coexisting ions did not interfere with the separation and determination of the analyte. The maximum static adsorption capacity of the sorbent at optimum conditions was found to be 32.5mgg(-1) while the time of 95% adsorption was less than 2min. The detection limit of present method was found to be 0.091microgg(-1), and the relative standard deviation (RSD) was lower than 3.0% (n=8). The method was successfully applied for the preconcentration of trace Sc(III) in the environmental samples with satisfactory results. PMID- 20006076 TI - New MSPQC-PLS method for the early clinic identification of commonly encountered Candida species. AB - The early clinic identification of commonly encountered Candida species became more important with the increasing human candidiasis. In this paper, a new MSPQC PLS (multi-channel series piezoelectric quartz crystal biosensor combined with partial least square) method was proposed for early identification of the most frequent Candida species encountered in human pathology. This method was based on these fact that (1) MSPQC method is a real-time monitoring method based on the sensitive frequency response to the change of electric parameters of the culture media caused by the growth of microorganisms; (2) various Candida species produce significantly different types of frequency curves in 1 or 2 days' culture period; (3) this difference can be identified by the partial least square technique. Using the proposed method, three species (Candida albicans, Candida glabrata, and Candida tropicalis) from a collection of 53 clinical strains of Candida, isolated from hospitalized patients, were identified with a classification rate of 98.1%. New proposed MSPQC-PLS method is simple, rapid and convenient to perform. It can identify clinical Candida species directly without passing through pure culture process. This will save identification time greatly. It could be popularized in clinical microbiology laboratories. PMID- 20006077 TI - Magnetic bead-based approach to monitoring of cigarette smoke-induced DNA oxidation damage and screening of natural protective compounds. AB - Cigarette smoking can damage DNA and induce spontaneous mutagenesis or carcinogenesis. Here, we describe a novel strategy for in situ monitoring of cigarette smoke-induced DNA oxidation damage and offer a method for screening natural compounds that protect DNA against tobacco smoke. The present protocol takes advantage of a fast and simple magnetic separation/mixing method and a highly sensitive chemiluminescence (CL) ELISA. The DNA immobilized on the magnetic beads was oxidized by the smoke in the absence or presence of natural compounds, and then oxidative DNA was conveniently held by magnetic force, whereas the complex tobacco smoke matrix and any remaining compounds were completely eliminated by extensive washing, and possible interferences were thus removed and oxidative damage was then sensitively monitored by CL ELISA. A library of 32 natural products was then screened and three were found to protect DNA from oxidative damage and thus may be promising compounds for the development of new drugs. Moreover, the protection effect of these three natural compounds against DNA oxidation damage was successfully classified by directly spiking them in the reference cigarettes. In addition, the potential to screen a mixture in a complex sample matrix, such as crude extracts, was also demonstrated, and hence the proposed technique can screen compounds within a complex matrix and enhance the screening throughput. PMID- 20006078 TI - Development of an on-line isotope dilution laser ablation inductively coupled plasma mass spectrometry (LA-ICP-MS) method for determination of boron in silicon wafers. AB - A method has been developed based on an on-line isotope dilution technique couple with laser ablation/inductively coupled plasma mass spectrometry (LA-ICP-MS), for the determination of boron in p-type silicon wafers. The laser-ablated sample aerosol was mixed on-line with an enriched boron aerosol supplied continuously using a conventional nebulization system. Upon mixing the two aerosol streams, the isotope ratio of boron changed rapidly and was then recorded by the ICP-MS system for subsequent quantification based on the isotope dilution principle. As an on-line solid analysis method, this system accurately quantifies boron concentrations in silicon wafers without the need for an internal or external solid reference standard material. Using this on-line isotope dilution technique, the limit of detection for boron in silicon wafers is 2.8x10(15)atomscm(-3). The analytical results obtained using this on-line methodology agree well with those obtained using wet chemical digestion methods for the analysis of p-type silicon wafers containing boron concentrations ranging from 1.0x10(16) to 9.6x10(18)atomscm(-3). PMID- 20006079 TI - Study the damage of DNA molecules induced by three kinds of aqueous nanoparticles. AB - In this paper, the interaction of DNA molecules with aqueous CdTe quantum dots (CdTe QDs), CdTe/SiO(2) composite nanoparticles (CdTe/SiO(2) NPs), and Mn-doped ZnSe quantum dots (Mn:ZnSe d-dots) was studied with ethidium bromide as a probe. The purpose of this work was to study the damage of DNA molecules induced by these three kinds of water-soluble nanoparticles. It was found that ionic strength, pH value and UV irradiation influenced the PL emission properties of CdTe QDs, CdTe/SiO(2) NPs and Mn:ZnSe d-dots, and also influenced the interaction of DNA molecules with them. Among the three kinds of nanoparticles, DNA molecules were most easily damaged by CdTe QDs whether in the dark or under UV irradiation. The CdTe/SiO(2) NPs led to much less DNA damage when compared with CdTe QDs, as a silica overcoating layer could isolate the QDs from the external environment. Mn:ZnSe d-dots as a new class of non-cadmium doped QDs demonstrated almost no damage for DNA molecules, which have great potentials as fluorescent labels in the applications of biomedical assays, imaging of cells and tissues, even in vivo investigations. PMID- 20006080 TI - Determination of trace iodide in iodised table salt on silver sulfate-modified carbon paste electrode by differential pulse voltammetry with electrochemical solid phase nano-extraction. AB - Electrochemical solid phase nano-extraction, a novel sample preparation technique, was used for the determination of trace iodide in iodised table salt based on the silver sulfate nanoparticle-modified carbon paste electrode. Electrochemical solid phase nano-extraction was realized in the exchange between the sulfate anion in nanoparticles and an iodide anion from aqueous solution. The released silver cation serves as the electrochemical probe for the determination of iodide. The extraction follows a Freundlich adsorption isotherm, and can be used in the detection of iodide in the concentration range 5.0x10(-12)-4.0x10( 9)M. The amount of iodide in iodised table salt was determined as 0.875+/ 0.002microg/g, which is about 2.5% of the addition amount of iodate with a relative deviation of 5.92% and a standard addition recovery of 90-110%. The large amounts of chloride and iodate did not interfere with the detection. PMID- 20006081 TI - UV-induced carbonyl generation with formic acid for sensitive determination of nickel by atomic fluorescence spectrometry. AB - UV-induced carbonyl generation with formic acid is used for gaseous sample introduction into an atomic fluorescence spectrometer for the determination of ultra-trace nickel. Compared with conventional carbonyl generation, no toxic gas CO is involved in this work, and volatile Ni(CO)(4) is generated with a single reagent formic acid under the irradiation of UV light (253.7nm, 15W). The reaction conditions, including reaction medium, UV irradiation time and reaction temperature, are optimized for the best signal. Under the optimized conditions, a limit of detection of 10ngL(-1) for nickel is obtained without any analyte-pre concentration, which is comparable to that using in situ trapping technique. Interferences from common transition metal ions, noble metal ions and mineral acids are also investigated. The proposed method is applied to the analysis of three certified reference materials and two organic acid samples for trace nickel, with analytical results in good agreement with certified values or those obtained by electrothermal atomic absorption spectrometry. This is a simple, fairly green and highly sensitive method for ultra-trace nickel determination. PMID- 20006082 TI - Discrimination of Ephedra plants with diffuse reflectance FT-NIRS and multivariate analysis. AB - The secondary metabolites of different Ephedra plants are various. Therefore, the discrimination of different Ephedra plants is significant. An objective, easy-to use, rapid and pollution-free approach is proposed for discriminating Ephedra plants of different species, habitats and picking times on the basis of diffuse reflectance Fourier transform near infrared spectroscopy (FT-NIRS) measurements and multivariate analysis. The Fourier transform near infrared diffuse reflectance spectra (NIRDRS) were acquired from 37 pulverized samples of Ephedra plants put in glass vials in the near infrared (NIR) region between 10000 and 4000cm(-1), averaging 64 scans per spectrum at a resolution of 4cm(-1). After spectra processing and data pre-processing, spectral data were analyzed respectively with three multivariate analysis techniques: discriminant analysis (DA), self-organizing map (SOM) and back-propagation artificial neural network (BP-ANN). The proposed method could distinguish not only the Ephedra plants of three species and two habitats but also the plants picked at different times of day without special sample treatment and the use of chemical reagents. The performance indexes of the DA model were 84.2-91.9% and the prediction accuracies of both the SOM and the BP-ANN models reached 93.3-100.0%. PMID- 20006083 TI - A highly sensitive HPLC method with automated on-line sample pre-treatment and fluorescence detection for determination of reboxetine in human plasma. AB - A fully automated, rapid and highly sensitive HPLC method with automated sample pre-treatment by column-switching system and fluorescence detection has been developed for the trace quantitative determination of the new antidepressant reboxetine (RBX) in human plasma. A simple pre-column derivatization procedure with 7-flouro-4-nitrobenzo-2-oxa-1,3-diazole (NBD-F) reagent was employed. Paroxetine (PXT) was used as an internal standard. Plasma samples containing both RBX and PXT, after filtration, were derivatized by heating with NBD-F in borate buffer of pH 8 at 70 degrees C for 30min. The derivatized plasma samples were injected into the HPLC system where an on-line sample clean up was achieved on the pre-treatment column (Co-sense Shim-pack MAYI-ODS) with a washing mobile phase (acetonitrile:2% acetic acid; 40:60, v/v) at a flow rate of 5mLmin(-1) for 1min. After an automated on-line column switching to the analytical Hypersil phenyl 120A column (250mmx4.6mm, 5microm), the separation of the derivatized RBX and PXT was performed using a mobile phase consisting of sodium acetate buffer (pH 3.5):tetrahydrofuran:acetonitrile (55:35:10, v/v/v) at a flow rate of 2.0mLmin(-1). The eluted derivatives were monitored by a fluorescence detector set at an excitation wavelength of 470nm and an emission wavelength of 530nm. Under the optimum chromatographic conditions, a linear relationship with good correlation coefficient (r=0.9995, n=5) was found between the peak area ratio of RBX to PXT and RBX concentration in the range of 2-500ngmL(-1), with limits of detection and quantification of 0.5 and 1.7ngmL(-1), respectively. The intra- and inter-day precisions were satisfactory; the relative standard deviations were 2.25 and 3.01% for the intra- and inter-day precisions, respectively. The accuracy of the method proved as the mean recovery values were 100.11+/-2.24% and 100.99+/-2.98% for the intra- and inter-day assay runs, respectively. The proposed method involved simple and minimum sample preparation procedure and short run-time (<12min) and therefore it can be applied to the routine therapeutic monitoring and pharmacokinetic studies of RBX. PMID- 20006084 TI - A modified acid digestion procedure for extraction of tungsten from soil. AB - Interest in tungsten occurrence and geochemistry is increasing due to increased use of tungsten compounds and its unknown biochemical effects. Tungsten has a complex geochemistry, existing in most environmental matrices as the soluble and mobile tungstate anion, as well as poly- and heteropolytungstates. Because the geochemistry of tungsten is substantially different than most trace metals, including the formation of insoluble species under acidic conditions, it is not extracted from soil matrices using standard acid digestion procedures. Therefore, the current work describes a modification to a commonly used acid digestion procedure to facilitate quantification of tungsten in soil matrices. Traditional soil digestion procedures, using nitric and hydrochloric acids with hydrogen peroxide yield <1 up to 50% recovery on soil matrix spike samples, whereas the modified method reported here, which includes the addition of phosphoric acid, yields spike recoveries in the 76-98% range. Comparison of the standard and modified digestion procedures on National Institute of Standards and Technology Standard Reference Materials yielded significantly improved tungsten recoveries for the phosphoric acid modified method. The modified method also produces comparable results for other acid extractable metals as the standard methods, and therefore can be used simultaneously for tungsten and other metals of interest. PMID- 20006085 TI - Fluorescence aptasensor based on competitive-binding for human neutrophil elastase detection. AB - To our knowledge, we report the first fluorescence aptasensor for detecting human neutrophil elastase (HNE) in homogeneous solution. The biosensor contains a short DNA scrambled sequence strand (SS) complementary to part of the aptamer sequence or the loop of molecular beacon (MB). The aptamer-HNE recognition event involves competition between the molecular beacon and loose HNE aptamer for the binding the short DNA strand. The new biosensor can detect as little as 0.34nM of HNE, and the response is linear in the tested concentration range of 0.34-68nM with the detection limit of 47pM. PMID- 20006086 TI - The influence of indoxyl sulfate and ammonium on the autofluorescence of human urine. AB - Despite biological variability the spectral characteristics of undiluted human urine show relatively low autofluorescence at short UV (250-300nm) excitation. However with dilution the fluorescence intensity remarkably increases. This paper examines the mechanisms behind this effect, by using excitation-emission matrices. Corrections for the inner filter effect were made for improved understanding of the spectral patterns. We focused on three major fluorophores (tryptophan, indoxyl sulfate and 5-hydroxyindole-3-acetate) that are excited at these wavelengths, and whose content in urine is strongly linked with various health conditions. Their fluorescence was studied both individually and in combinations. We also examined the effect of ammonium on the fluorescence of these major fluorophores individually and in combinations. Through these studies we have identified the leading effects that reduce the UV fluorescence, namely higher concentration of indoxyl sulfate producing the inner filter effect and concentration quenching and quenching of fluorophores by ammonium. This result will assist in broader utilisation of UV fluorescence in medical diagnostics. PMID- 20006088 TI - Determination of sodium, potassium, calcium, magnesium, zinc, and iron in emulsified egg samples by flame atomic absorption spectrometry. AB - In this study, oil-in-water formulations were optimized to determine sodium, potassium, calcium, magnesium, zinc, and iron in emulsified egg samples by flame atomic absorption spectrometry (FAAS). This method is simpler and requires fewer reagents when compared with other sample pre-treatment procedures and allows the calibration to be carried out using aqueous standards. Different oily phases such as corn oil, decyl oleate and octyl stearate were tested, as well as Tween 80, Triton X-100 and Triton 114 were analyzed as surfactants. The optimum type and proportion of formulations were determined and their use depended on the element studied. The emulsion preparation was performed by a conventional method that involves mixing both phases at 60 degrees C by magnetic stirring and phase inversion to change the water-to-oil ratio by increasing the volume of the surfactant-water external phase and correspondingly decreasing the volume of internal phase. The accuracy of the method was further confirmed by determining the metals in a whole egg powder CRM and recoveries ranged from 97.5% for Mg to 102.2% for Na, with relative standard deviations lower than 2.3%. The precision of the procedures was determined through repeatability (intra-day precision) and intermediate precision (inter-day). The repeatability presented RSD values lower than 4.2%. The intermediate precision was evaluated using the RSD and F-test. The RSD values to intermediate precision was lower than 5.3% and the computed F values were lower than tabulated F-values, indicating no significant difference between the results obtained on different days. The proposed method including, sample emulsification for subsequent metal determination for FAAS, has proved to be sensitive, reproducible, simple and economical. PMID- 20006087 TI - Electrocatalytic oxidation of phytohormone salicylic acid at copper nanoparticles modified gold electrode and its detection in oilseed rape infected with fungal pathogen Sclerotinia sclerotiorum. AB - Salicylic acid (SA) is a biological substance that acts as a phytohormone and plays an important role in signal transduction in plants. It is important to accurately and sensitively detect SA levels. A gold electrode modified with copper nanoparticles was used to assay the electrocatalytic oxidation of salicylic acid. It was found that the electrochemical behavior of salicylic acid was greatly improved at copper nanoparticles, indicating that anodic oxidation could be catalyzed at copper nanoparticles. And the pH had remarkable effect on the electrochemical process, a very well-defined oxidation peak appeared at pH 13.3 (0.2M NaOH). The kinetics parameters of this process were calculated and the heterogeneous electron transfer rate constant (k) was determined to be 1.34x10( 3)cms(-1), and (1-alpha)n(alpha) was 1.22. The gold electrode modified with copper nanoparticles could detect SA at a higher sensitivity than common electrodes. The electrode was used to detect the SA levels in oilseed rape infected with the fungal pathogen Sclerotinia sclerotiorum. The results showed that the SA concentration reached a maximum during the 10th-25th hours after infection. This result was very similar to that determined by HPLC, indicating that the gold electrodes modified with copper nanoparticles could be used as salicylic acid sensors. PMID- 20006089 TI - A highly selective nanogold-aptamer catalytic resonance scattering spectral assay for trace Hg(2+) using HAuCl(4)-ascorbic acid as indicator reaction. AB - Single strand DNA (ssDNA) was used to modify nanogold to obtain a nanogold aptamer resonance scattering (RS) probe (NGssDNA) for Hg(2+), based on the formation of stable thymine-Hg(2+)-thymine (T-Hg(2+)-T) mismatches and aggregation of the released nanogold particles. After removing the aggregated particles by filtrate membrane, the excess NGssDNA in the filtration solution exhibit catalytic effect on the gold particle reaction between HAuCl(4) and ascorbic acid (AA) that appear as RS peak at 596nm. When Hg(2+) concentration increased, the RS intensity at 596nm decreased. The decreased intensity is linear to Hg(2+) concentration in the range of 0.00008-0.888ng/mL Hg(2+), with detection limit of 0.000034ng/mL. The nanogold-aptamer catalytic RS assay was applied to determination of Hg(2+) in water with satisfactory results. PMID- 20006090 TI - Application of ionic liquids based microwave-assisted extraction of three alkaloids N-nornuciferine, O-nornuciferine, and nuciferine from lotus leaf. AB - The application of ionic liquids based microwave-assisted extraction (ILMAE) was successfully developed for extracting three alkaloids N-nornuciferine, O nornuciferine, and nuciferine from lotus leaf. Seven kinds of 1-alkyl-3 methylimidazolium with different cations and anions were investigated in this work and 1.0M 1-hexyl-3-methylimidazolium bromide ([C(6)MIM]Br) solution was selected as solvent. In addition, the microwave parameters including irradiation power, extraction time and solid-liquid ratio were optimized. Compared with the regular MAE and conventional heat-reflux extraction (HRE), the proposed approach exhibited higher efficiency (0.9-43.7% enhanced) and shorter extraction time (from 2h to 2min), which indicated ILMAE was an efficient, rapid and simple sample preparation technique. Moreover, the proposed method was validated by the linearity, reproducibility, and recovery experiments. Good linearity was observed with the regression coefficients (r(2)) between 0.9998 and 0.9999. The recoveries of all methods were in the range of 94.6% and 105.5% with RSD lower than 6.6%, which indicated that the proposed method was credible. PMID- 20006091 TI - Immobilization of enzyme on detonation nanodiamond for highly efficient proteolysis. AB - Immobilization of enzyme on detonation nanodiamond (dND, 3-10nm) and its application for efficient proteolysis have been demonstrated. By evaluation of the Michaelis constant (K(m)) and maximum velocity (V(max)) of immobilized enzyme, its activity was not impaired significantly by immobilization. And enzyme immobilized on dNDs exhibited much better thermal and chemical stabilities than its free counterpart and maintained high activity even after 10 times reuse. The efficient proteolysis by trypsin immobilized on dNDs (dND-trypsin) is demonstrated with the digestion of myoglobin (or other model protein) in a short time (5min). Large numbers of identified peptides obtained by dNDs-trypsin enable a higher degree of sequence coverage and more positive identification of proteins than those obtained by in-solution digestion and the commercial immobilized trypsin beads, respectively. Moreover, immobilization of peptide-N-glycosidase F (PNGase F) on dNDs was realized and resulted in faster sequential glycosidase digestion of glycopeptides in less than 10min. PMID- 20006092 TI - Preparation of a new Cd(II)-imprinted polymer and its application to determination of cadmium(II) via flow-injection-flame atomic absorption spectrometry. AB - A new cadmium(II)-imprinted polymer based on cadmium(II) 2,2'-{ethane-1,2 diylbis[nitrilo(E)methylylidene]} diphenolate-4-vinylpyridine complex was obtained via suspension polymerization. The beads were used as a minicolumn packing for flow-injection-flame atomic absorption spectrometry (FI-FAAS) determination of cadmium(II) in water samples. Sorption effectiveness was optimal within pH range of 6.6-7.7. Nitric acid, 0.5% (v/v) was used as eluent. Fast cadmium(II) sorption by the proposed material enabled to apply sample flow rates up to 10mLmin(-1) without loss in sorption effectiveness. Enrichment factor (EF), concentration efficiency (CE) and limit of detection (LOD, 3sigma) found for 120 s sorption time were 117, 39.1min(-1) and 0.11microgL(-1), respectively. Sorbent stability was proved for at least 100 preconcentration cycles (RSD=2.9%). When compared to non-imprinted polymer the new Cd(II)-imprinted polymer exhibited improved selectivity towards cadmium(II) against other heavy metal ions, especially Cu(II) and Pb(II), as well as light metal ions. Accuracy of the method was tested for ground water and waste water certified reference materials and fortified water. The method was applied to Cd(II) determination in natural water samples. PMID- 20006093 TI - Comparison of tetraethylborate and tetraphenylborate for selenite determination in human urine by gas chromatography mass spectrometry, after headspace solid phase microextraction. AB - Two different derivatizing reagents were tested for the development of a fast and sensitive method for the determination of selenites (Se(IV)) in human urine. The reagents were sodium tetraethylborate (NaBEt(4)) and tetraphenylborate (NaBPh(4)), respectively, and the procedure is based on in situ derivatization of selenites in aqueous medium. Selenite ions are converted to diethylselenide (DESe) or diphenylselenide (DPhSe) and subsequently collected from the headspace by solid phase microextraction using a silica fiber coated with polydimethylsiloxane (HS-SPME). Finally, they are quantitated by GC/MS in SIM mode. Ethylation over phenylation was proved preferable for the headspace extraction because of the higher volatility of the diethyl-derivative of selenites. The optimization of the HS-SPME conditions was performed both in aqueous and urinary solutions. Under the optimum conditions for HS-SPME, the gas chromatographic conditions were also optimized. Between the two alkylation reagents tetraethylborate was proved more efficient and the quantitation was satisfactory. Aqueous certified reference materials were analyzed to evaluate the accuracy of the method. The precision of the method was 4.2% and the calculated detection limit was 0.05microgL(-1) for human urine. PMID- 20006094 TI - Design of a new electrogenerated polyquinone film substituted with glutathione. Towards direct electrochemical biosensors. AB - We developed a method to graft a tripeptide (glutathione) onto 5-hydroxy-1,4 naphthoquinone, an electropolymerizable molecule. The resulting thin conducting polymer presents a well-defined and stable electroactivity in neutral buffered solution, due to the embedded quinone group, and is able to covalently graft amino-modified DNA probe strands. It is shown that the bioelectrode presents positive current change following DNA hybridization. This makes a "signal-on" direct electrochemical DNA sensor. The results were obtained with low target concentration (50nM) and the selectivity is excellent as a single-mismatch sequence can be discriminated from the full-complementary target. PMID- 20006095 TI - Single interface flow system with potentiometric detection for the determination of nitrate in water and vegetables. AB - In this work a single interface flow system (SIFA) with potentiometric detection was for the first time implemented and applied to the determination of nitrate in waters and plant extracts. The analytical potential of the SIFA system was exploited not only to transport the sample towards detection but also to carry out, in a reproducible and automated way, the tasks associated with sample pre treatment, namely ionic strength, pH adjustment and interfering species suppression. The advantageous aspects of combining a SIFA system with potentiometry with enhanced simplicity, ease of implementation and automation were further discussed and emphasised. The obtained results showed relative deviations lower than 5%, for both types of samples, with sampling rates of about 40h(-1). In addition, an innovative and straightforward process for constructing plastic membrane ion selective electrodes with a tubular configuration able to be coupled to flow-based analytical systems is also proposed. The developed approach, consisting of assembling the electrode inside a flow tubing connector is very simple to implement, robust, particularly adequate to be combined with flow methodologies and maintains all dynamic and analytical characteristics exhibited by previous assembling processes. PMID- 20006096 TI - Interfacing on-line solid phase extraction with monolithic column multisyringe chromatography and chemiluminescence detection: An effective tool for fast, sensitive and selective determination of thiazide diuretics. AB - A new, multisyringe flow injection set-up has been developed for the completely automated determination of trace thiazide compounds with diuretic action in different types of samples. The proposed instrumental set-up exploits for the first time, a low pressure on-line solid phase extraction-liquid chromatography chemiluminescence detection method. This novel combination of sample treatments in flow systems expands the current applicability of low pressure liquid chromatography due to the isolation/preconcentration of the target compounds, besides high selectivity and sensitivity. For the determination of three thiazide compounds named hydroflumethiazide, furosemide and bendroflumethiazide, the proposed set-up provided with the preconcentration of only 1mL of sample, limits of detection of 3, 60 and 40microgL(-1), respectively. Furthermore wide linear dynamic ranges of 6-4000, 140-20,000 and 90-40,000microgL(-1), respectively, were obtained. Besides of this, a high injection throughput of 12h(-1) was also achieved. As in sports, thiazide diuretics are prohibited substances, the proposed method has been applied to their determination in urine samples. Furthermore the potential of the proposed method as a fast-screening approach for emerging contaminants in waters has been also tested by applying it to well water and leachates from a solid waste landfill. PMID- 20006097 TI - A multisyringe flow injection Winkler-based spectrophotometric analyzer for in line monitoring of dissolved oxygen in seawater. AB - An integrated analyzer based on the multisyringe flow injection analysis approach is proposed for the automated determination of dissolved oxygen in seawater. The entire Winkler method including precipitation of manganese(II) hydroxide, fixation of dissolved oxygen, dissolution of the oxidized manganese hydroxide precipitate, and generation of iodine and tri-iodide ion are in-line effected within the flow network. Spectrophotometric quantification of iodine and tri iodide at the isosbestic wavelength of 466nm renders enhanced method reliability. The calibration function is linear up to 19mgL(-1) dissolved oxygen and an injection frequency of 17 per hour is achieved. The multisyringe system features a highly satisfying signal stability with repeatabilities of 2.2% RSD that make it suitable for continuous determination of dissolved oxygen in seawater. Compared to the manual starch-end-point titrimetric Winkler method and early reported automated systems, concentrations and consumption of reagents and sample are reduced up to hundredfold. The versatility of the multisyringe assembly was exploited in the implementation of an ancillary automatic batch-wise Winkler titrator using a single syringe of the module for accurate titration of the released iodine/tri-iodide with thiosulfate. PMID- 20006098 TI - Voltammetric studies on the potent carcinogen, 7,12-dimethylbenz[a]anthracene: Adsorptive stripping voltammetric determination in bulk aqueous forms and human urine samples and detection of DNA interaction on pencil graphite electrode. AB - 7,12-Dimethylbenz[a]anthracene (DMBA), is a widely studied polycyclic aromatic hydrocarbon that has long been recognized as a very potent carcinogen. Initially, the electrochemical oxidation of DMBA at the glassy carbon and pencil graphite electrodes in non-aqueous media (dimethylsulphoxide with lithium perchlorate) was studied by cyclic voltammetry. DMBA was irreversibly oxidized in two steps at high positive potentials, resulting in the ill-resolved formation of a couple with a reduction and re-oxidation wave at much lower potentials. Special attention was given to the use of adsorptive stripping voltammetry together with a medium exchange procedure on disposable pencil graphite electrode in aqueous solutions over the pH range of 3.0-9.0. The response was characterized with respect to pH of the supporting electrolyte, pre-concentration time and accumulation potential. Using square-wave stripping mode, the compound yielded a well-defined voltammetric response in acetate buffer, pH 4.8 at +1.15V (vs. Ag/AgCl) (a pre-concentration step being carried out at a fixed potential of +0.60V for 360s). The process could be used to determine DMBA concentrations in the range 2-10nM, with an extremely low detection limit of 0.194nM (49.7ngL(-1)). The applicability to assay of spiked human urine samples was also illustrated. Finally, the interaction of DMBA with fish sperm double-stranded DNA based on decreasing of the oxidation signal of adenine base was studied electrochemically by using differential pulse voltammetry with a pencil graphite electrode at the surface and also in solution. The favorable signal-to-noise characteristics of biosensor resulted in low detection limit (ca. 46nM) following a 300-s interaction. These results displayed that the electrochemical DNA-based biosensor could be used for the sensitive, rapid, simple and cost effective detection of DMBA-DNA interaction. PMID- 20006099 TI - Batch and hydrodynamic monitoring of vitamin C using novel periodate selective sensors based on a newly synthesized Ni(II)-Schiff bases complexes as a neutral receptors. AB - A highly selective membrane electrodes based on a two newly synthesized nickel (II) Schiff bases, [NiL(1)] and [NiL(2)] where L(1) and L(2) are N,N/bis(salicylaldehyde)4,5-dimethyl-1,2-phenylenediamine (H(2)L(1)) and N,N/bis(salicylaldehyde)4,5-dichloro-1,2-phenylenediamine (H(2)L(2)) were used as a neutral carrier ionophores for static and hydrodynamic potentiometric mode of operations for the determination of periodate. Under static mode of operation, the sensors displayed a near-Nernstian slope of -66.1+/-0.8 and -59.9+/-1.1mV decade(-1) of activity and detection limits to 5.2x10(-6) and 7.3x10(-6)molL(-1) for the sensors based on [NiL(1)] and [NiL(2)], respectively. Under hydrodynamic mode of operation (FIA), the slope of the calibration plot, limit of detection, and working linear range were -71.1mV decade(-1) of activity, 7.3x10(-6) and 1.0x10(-5) to 1.0x10(-3)molL(-1), respectively. The response time of the sensors in whole concentration ranges was very short (<10s). The response of the sensors was independent on the pH range of 3-8. A tubular version was further developed and coupled to a flow injection system for ascorbic acid (AA) determination in beverages and pharmaceutical preparations. This approach was achieved by selecting a 50-cm reactor and an overall flow of 3mLmin(-1), and injecting volume 100microL of AA standards in a 1.0x10(-4)molL(-1) IO(4)(-) solution. Under these conditions, a linearity range of 2-13microgmL(-1), with a slope of 4.97mV (mg/L)( 1) (r(2)=0.9995), detection limit 0.9mgL(-1) and a reproducibility of +/-1.1mV (n=5) was recorded. This simple and inexpensive flow injection analysis manifold, with a good potentiometric detector, enabled the analysis of approximately 50 samples h(-1) without requiring pretreatment procedures. An average recovery of 98.8% and a mean standard deviation of 1.3% were obtained. PMID- 20006100 TI - A sensitive and selective method for determination of gold(III) based on electrothermal atomic absorption spectrometry in combination with dispersive liquid-liquid microextraction using dicyclohexylamine. AB - A combined method with dispersive liquid-liquid microextraction (DLLME) and electrothermal atomic absorption spectrometry (ETAAS) has been developed for determining gold(III). Dicyclohexylamine, a new extractant for gold(III), showed excellent performance in DLLME. Acetone was indispensable to the quantitative extraction of gold(III), contributing to decrease in hydration, decrease in the difference in the dielectric constants between the supernatant phase and the sedimented phase, and dissolution of a part of chloroform as an extraction solvent to the supernatant phase as well as improvement of dipersibility. In DLLME using a mixture of 1.0mL of acetone and 100microL of chloroform containing 50mmolL(-1) of dicyclohexylamine, gold(III) could be extracted selectively and effectively from 8mL of a sample solution in the presence of iron(III), cobalt(II), nickel(II), copper(II), palladium(II), and platinum(IV) at pH 1. The extracted gold(III) was determinable by ETAAS; the detection limit was 0.002microgL(-1) (three times the standard deviation of the blank values, n=8) as a gold(III) concentration in 8mL of sample solution. The proposed method was applicable to the determination of gold in platinum metal and its alloy as well as effluent without any interference by the matrices. PMID- 20006101 TI - The porous CuO electrode fabricated by hydrogen bubble evolution and its application to highly sensitive non-enzymatic glucose detection. AB - The porous Cu film was deposited on a Pt/Ti/Si substrate by electrochemical deposition accompanied by hydrogen evolution at very high current densities. CuO films with similar morphologies were obtained by subsequent annealing of the porous copper films. The morphology, composition and structure of the porous Cu and porous CuO were investigated by FE-SEM, EDS and XRD methods. The complete transformation of Cu to CuO after annealing was indicated. The sensing performances of the porous CuO film were evaluated in alkaline solution with the porous CuO film showing a wide linearity range from 1microM to 2.5mM with sensitivity of 2.9mAcm(-2)mM(-1), and detection limit of 0.14microM. The sensor showed good selectivity to conventional intermediates such as AA and UA and long term stability. PMID- 20006102 TI - Fabrication and evaluation of chiral monolithic column modified by beta cyclodextrin derivatives. AB - Based on combination of chiral recognition ability of beta-cyclodextrin (beta-CD) derivatives and flexibility of monolithic material, a series of chiral stationary phases (CSPs) were prepared by the immobilization of beta-CD and three of its derivatives to the epoxy-activated poly(glycidyl methacrylate-co-ethylene dimethacrylate) monolith under mild condition. Immobilization condition for the connection reaction by different functional groups and bonding ways was studied to obtain good enantiomer selectivity. Prepared CSPs were evaluated by separating racemic mixtures of eight amino acids and two chiral drugs with capillary electrochromatography (CEC). PMID- 20006103 TI - Electrochemical immunoassay of hepatitis B surface antigen by the amplification of gold nanoparticles based on the nanoporous gold electrode. AB - We describe herein the combination of electrochemical immunoassay using nanoporous gold (NPG) electrode with horseradish peroxidase (HRP) labeled secondary antibody-gold nanoparticles (AuNPs) bioconjugates for highly sensitive detection of protein in serum. The electroactive product of o-phenylenediamine (OPD) oxidized with H(2)O(2) catalyzed by HRP was reduced in the Britton-Robinson (BR) buffer and the peak current of which was used to determine the concentration of antigen (Ag) in the analyte. The active surface area of NPG electrode was larger than that of a bare flat one. The presence of AuNPs enhanced the immobilized amount of HRP labeled antibody (Ab), which improved the sensitivity of the immunoassay when used as the secondary antibodies. As a result of these two combined effects, the sensitivity of the immunoassay for the determination of target protein was increased significantly. Using hepatitis B surface antigen (HBsAg) as a model, we demonstrate a dose response in the range of 0.01-1.0ng/mL with a detection limit of 2.3pg/mL. Analytical results of several human serum samples obtained using the developing technique are in satisfactory agreement with those given by enzyme-linked immune-absorbent assays (ELISA). In addition, the technique was about 100 times more sensitive in the detection of HBsAg than ELISA. All these demonstrated the feasibility of the present immunoassay method for clinical diagnosis. PMID- 20006104 TI - Dual-sensitive probe 1-imidazole-2-(5-benzoacridine)-ethanone for the determination of amines in environmental water using HPLC with fluorescence detection and online atmospheric chemical ionization-mass spectrometry identification. AB - Dual-sensitive probe of 1-imidazole-2-(5-benzoacridine)-ethanone (IBAE) for the determination of free amines with fluorescence detection and online highly sensitive atmospheric chemical ionization-mass spectrometry identification (APCI MS) has been developed. 2-(Benzoacridine)-5-acetic acid (BAAA) reacts with coupling agent N,N'-carbonyldiimidazole (CDI) to form a highly activated amide intermediate 1-imidazole-2-(5-benzoacridine)-ethanone (IBAE), which is dual sensitive probe. The amide intermediate (IBAE) reacts preferably with amines in dimethylformamide (DMF) solvent to give the high yields of derivatives. IBAE amine derivatives are not only sensitive to fluorescence but also to MS ionizable efficiency. The percent ionization delta values change from 0 to 57.32% in aqueous acetonitrile and from 0 to 62.14% in aqueous methanol. The relative standard deviations of the peak areas with fluorescence detection for each amine are <1.24% (40ng/ml, n=6). The fluorescence detection limits (at a signal-to noise ratio of 3) are in the range of 0.15-0.50ng/ml. The online APCI-MS detection limits are in the range of 2.07-8.51ng/ml (at a signal-to-noise ratio of 3). Therefore, the facile IBAE intermediate derivatization allowed the development of a highly sensitive and specific method for the quantitative analysis of trace levels of amines in environmental water. PMID- 20006105 TI - Cuprous oxide nanospheres as probes for light scattering imaging analysis of live cells and for conformation identification of proteins. AB - A facile solution-phase synthesis route of highly uniform Cu(2)O nanospheres (Cu(2)O NPs) with the size of 57.7+/-4.7nm was developed, and then the nanoparticles were applied to live cell imaging under a common dark-field microscope. Starting from copper(II) salts, the synthesis of Cu(2)O NPs was made in the presence of cetyltrimethylammonium bromide (CTAB) by reducing the copper(II) with sodium borohydride (NaBH(4)) in aqueous medium and by aging process in the air. Monitoring of morphology evolution process of Cu(2)O NPs with scanning electron microscopy (SEM) and measuring of the UV-visible spectra showed that the synthesis of Cu(2)O NPs follows the reduction-oxidation coupled process of Cu(2+) into Cu(0) species at first and then the resulted Cu(0) species into Cu(2)O NPs in the air. Light scattering (LS) features have been measured with a common spectrofluorometer and a common dark-field microscope, and it was found that the as-prepared Cu(2)O NPs display strong blue scattering light and can be applied for cell imaging. If incubated with human bone marrow neuroblastoma, transferrin-conjugated Cu(2)O NPs can get into the cells and show strong pure blue light in cytoplasm. Further investigations showed that the Cu(2)O NPs could be applied for probes for conformation of proteins. PMID- 20006106 TI - Investigation of the effects of 24 bio-matrices on the LC-MS/MS analysis of morinidazole. AB - This study compares and evaluates the effect of various matrices on liquid chromatography (LC) coupled with tandem mass spectrometry (MS/MS) analysis. Permanent post-column infusion (PCI) was used to quantify matrix effects. In this way, the suppressed or enhanced signal of the target material resulting from different co-eluting matrix components could be assessed. Twenty-four biological samples from in vivo and in vitro experiments were selected for this study. In addition, 7 sample components were further analyzed after sample preparation by protein precipitation. Multiple regression analysis was used to investigate the collinear relationship between matrix effects and co-eluted components at different time intervals. We found that salt was the dominant factor which impacted changes in signal detection. In order to eliminate it, we used ammonium formate as a modifier of the mobile phase which resulted in charge-state redistribution profiles so that a homogeneous matrix formed. By employing pulse gradient chromatography in the presence of 5mM ammonium formate, favorable improvements of enhanced signal intensity and reduced matrix effects were obtained. These experiments also indicated the feasibility of using analogue IS during bio-analysis which contributed to an overall faster assay that would be suitable for drug discovery and development purposes. PMID- 20006107 TI - Development and validation of a quantitative method for the selective determination of tin species in tin octoate by differential pulse polarography. AB - Tin octoate is used as a catalyst in the synthesis of polydimethylsiloxane (PDMS), a room temperature vulcanizing (RTV) silicone rubber. This rubber is largely used in the medical field due to its great biocompatibility. In this framework, a high-speed and costless analytical method for the determination of stannic ions, Sn(IV), in the presence of stannous ions, Sn(II), has been developed. The separation of these two ions was carried out using differential pulse polarography (DPP). For this purpose, the tin species contents in the catalyst is quantitatively extracted under inert condition to avoid any changes in the ratio Sn(IV)/Sn(II). Polarography showed well-shaped oxidation and reduction peaks respectively at -650 and -860mV for stannous ions. The peak of the stannic ion was well separated and appeared at -1210mV. Many parameters such as extraction process, extraction time, pH, chelating agents and polarographic conditions were optimized. We have also demonstrated that no oxidation of the stannous ions occurred during the sample preparation. The dosing range considered in this study extends between 10 and 40mug/mL, corresponding to 6.8% and 27.2% of the degradation product (Sn(IV)) in the catalyst, regarding to the sampling. Finally this method was successfully validated using the total error concept. PMID- 20006109 TI - Two analyte calibrations from the transient response of a single potentiometric sensor employed with the SIA technique. AB - Simultaneous quantification of Cd(2+) and Pb(2+) in solution has been correctly targeted using the kinetic information from a single non-specific potentiometric sensor. Dual quantification was accomplished from the complex information in the transient response of an electrode used in a Sequential Injection Analysis (SIA) system and recorded after step injection of sample. Data was firstly preprocessed with the Discrete Wavelet Transform (DWT) to extract significant features and then fed into an Artificial Neural Network (ANN) for building the calibration model. DWT stage was optimized regarding the wavelet function and decomposition level, while the ANN stage was optimized on its structure. To simultaneously corroborate the effectiveness of the approach, two different potentiometric sensors were used as study case, one using a glass selective to Cd(2+) and another a PVC membrane selective to Pb(2+). PMID- 20006108 TI - Retention of arsenate using genetically modified coryneform bacteria and determination of arsenic in solid samples by ICP-MS. AB - A novel method for the retention of arsenate [As(V)] combining time-controlled solid-phase extraction with living bacterial biomass is presented. As(V) retention was carried out by exposing the extractant, consisting of a living double-mutant of Corynebacterium glutamicum strain ArsC1-C2, to the sample for a retention time of 1-7min, before the arsenic distribution equilibrium between the sample solution and the extractant was established. The amount of As(V) retained in the biomass was measured by inductively coupled plasma-mass spectrometry (ICP MS) after the sample had been treated with nitric acid. A theoretical model of the retention process was developed to describe the experimental retention-time profiles obtained with the bacterial cells. This relationship provided a feasible quantification of the retention process before steady-state was reached, providing that the agitation conditions and the retention time had been controlled. An analytical procedure for the retention/quantification of As(V) was then developed; the detection limit was 0.1 ng As(V)mL(-1) and the relative standard deviation 2.4-3.0%. The maximum effective retention capacity for As(V) was about 12.5mgAs(g biomass)(-1). The developed procedure was applied to the determination of total arsenic in coal fly ash, using a sample that had undergone oxidative pre-treatment. PMID- 20006110 TI - Simultaneous kinetic determination of 3-hydroxybutyrate and 3-hydroxyvalerate in biopolymer degradation processes. AB - A new kinetic method is proposed for the simultaneous determination of 3 hydroxybutyrate (3HB) and 3-hydroxyvalerate (3HV) based on the different rate of the 3-hydroxybutyrate dehydrogenase-catalysed reactions of these compounds with coenzyme NAD(+). A flow injection system with two reactors of immobilised 3 hydroxybutyrate dehydrogenase and dual detection is used. The concentrations of NADH produced after two different reaction times are measured by fluorometry or spectrophotometry and multivariate linear calibration is applied for quantification. Concentrations of 3HB and 3HV between 1x10(-6) and 1x10(-4)M can be determined at an average sampling frequency of 20h(-1). In contrast to usual methods, the proposed here makes possible the discrimination of 3HB and 3HV without previous separation so that usual extraction with chlorinated solvents and/or chromatographic separation is not required. The method is of interest in a wide variety of fields concerning PHAs, as it can provide information on the degradation rate and mechanism, composition and structure of these polymers. Its applicability has been proved through the determination of 3HB and 3HV in the digests of some chemically degraded commercial PHAs. PMID- 20006111 TI - Arsenic species in certified reference material MURST-ISS-A2 (Antarctic krill). AB - Arsenic compounds were quantified in the certified reference material MURST-ISS A2 (Antarctic krill) using HPLC/ICPMS. The data should prove useful for assessing the accuracy of arsenic speciation procedures. PMID- 20006112 TI - Analysis of commercial beverage products by size exclusion chromatography coupled with UV-vis absorbance detection and dynamic surface tension detection. AB - Multidimensional analysis of instant coffee and barley beverage samples using size exclusion chromatography (SEC) combined with a dynamic surface tension detector (DSTD) and a UV-vis absorbance detector (UV) is reported. A unique finding of this study was the action of the tetrabutylammonium (TBA) cation as a modifying agent (with bromide as the counter anion) that substantially increased the surface pressure signal and sensitivity of many of the proteins in the chromatographically separated samples. The tetrabutylammonium bromide (TBAB) enhancement of the surface pressure signal was further investigated by studying the response of 12 commercial standard proteins (alpha-lactalbumin, beta lactoglobulin, human serum albumin (HSA), albumin from chicken egg white (OVA), bovine serum albumin (BSA), hemoglobin, alpha-chymotrypsinogen A, cytochrome C, myoglobin, RNase A, carbonic anhydrase, and lysozyme) in buffer performed using flow injection analysis (FIA) coupled with the DSTD with and without various concentrations of TBAB. The FIA-DSTD data show that 1mM TBAB enhances sensitivity of HSA detection, by lowering the limit of detection (LOD) from 2mg/mL to 0.1mg/mL. Similarly, the LOD for BSA was reduced from 1mg/mL to 0.2mg/mL. These FIA-DSTD experiments allowed the detection conditions to be optimized for further SEC-UV/DSTD experiments. Thus, the SEC-UV/DSTD system has been optimized and successfully applied to the selective analysis of surface-active protein fractions in a commercial instant coffee sample and in a soluble barley sample. The complementary selectivity of using the DSTD relative to an absorbance detector is also demonstrated. PMID- 20006114 TI - Cu nanoparticles incorporated polypyrrole modified GCE for sensitive simultaneous determination of dopamine and uric acid. AB - Cu nanoparticles have been electrochemically incorporated polypyrrole film that was used for modification of the glassy carbon electrode surface. The performance of the electrode has been characterized by cyclic voltammetry and atomic force microscopy. The electrode has shown high electrocatalytic activity towards the oxidation of dopamine (DA) and uric acid (UA) simultaneously in a phosphate buffer solution (pH 7.00). The electrocatalytic oxidation currents of UA and DA were found linearly related to concentration over the range 1x10(-9) to 1x10(-5)M for UA and 1x10(-9) to 1x10(-7)M for DA using DPVs method. The detection limits were determined as 8x10(-10)M (s/n=3) for UA and 8.5x10(-10)M (s/n=3) for DA at a signal-to-noise ratio of 3. PMID- 20006113 TI - Sorption of As(V) from waters using chitosan and chitosan-immobilized sodium silicate prior to atomic spectrometric determination. AB - A natural biosorbent containing amine functional groups, chitosan, and a novel sorbent, chitosan-immobilized sodium silicate, were prepared and utilized for the selective sorption of As(V) from waters prior to its determination by atomic spectrometric techniques, namely, hydride generation atomic absorption spectrometry (HGAAS) and inductively coupled plasma mass spectrometry (ICP-MS). Chitosan was synthesized from chitin and sodium silicate was used as the immobilization matrix due to its straightforward synthesis. Through sequential sorption studies, it was shown that chitosan-immobilized sodium silicate has exhibited a better chemical stability than the chitosan itself which demonstrates the advantage of immobilization method. Both chitosan and chitosan-immobilized sodium silicate were shown to selectively adsorb As(V), arsenate, from waters at pH 3.0 at which neither chitin nor sodium silicate displayed any sorption towards As(V). The sorption of arsenate by chitosan is supposed to have electrostatic nature since pH of 3.0 is both the point at which the amino groups in chitosan are protonated and also the predominant form of As(V) is H(2)AsO(4)(-). A pre oxidation step is required if both As(III) and As(V) are to be determined. Desorption from the sorbents was realized with 1.0% (w/v) l-cysteine prepared in a pH 3.0 solution adjusted with HCl. Among the possible interfering species tested, only Te(IV) and Sb(III) were shown to cause a decrease in the sorption capacity especially at high interferant concentrations. High concentrations of Sb(III) also resulted in gas phase interference during hydride generation. The validity of the method was checked both via spike recovery experiments and also through the analysis of a standard reference material. Spike recovery tests were carried out with four different types of water; namely, ultra-pure, bottled drinking, tap, and sea water; and percent recovery values were found to be 114 (+/-4), 112 (+/-2), 43 (+/-4), and 0 (+/-1), respectively. It was concluded that the proposed methodology can be applied efficiently to low-to-medium ionic strength solutions, such as most drinking waters. The accuracy of the method was additionally investigated through the analysis of a standard reference material and a good correlation was found between the determined (26.6+/-2.4microgL(-1)) and the certified (26.67microgL(-1)) value. PMID- 20006115 TI - The reference materials syndrome. AB - This paper examines specific cases in the literature where analysts using spectroscopic instrumentation report elemental concentrations that agree with information values reported in reference material certificates that are subsequently found to be incorrect. PMID- 20006116 TI - Enantiomer analysis of chiral carboxylic acids by AIE molecules bearing optically pure aminol groups. AB - Pure enantiomers of carboxylic acids are a class of important biomolecules, chiral drugs, chiral reagents, etc. Analysis of the enantiomers usually needs expensive instrument or complex chiral receptors. However, to develop simple and reliable methods for the enantiomer analysis of acids is difficult. In this paper, chiral recognition of 2,3-dibenzoyltartaric acid and mandelic acid was first carried out by aggregation-induced emission molecules bearing optically pure aminol group, which was easily synthesized. The chiral recognition is not only seen by naked eyes but also measured by fluorophotometer. The difference of fluorescence intensity between the two enantiomers of the acids aroused by the aggregation-induced emission molecules was up to 598. The chiral recognition could be applied to quantitative analysis of enantiomer content of chiral acids. More chiral AIE amines need to be developed for enantiomer analysis of more carboxylic acids. PMID- 20006117 TI - How to reduce prescribing errors. PMID- 20006118 TI - Universal health coverage: access to what? PMID- 20006119 TI - An opportunity to improve health in the European Union. PMID- 20006121 TI - Climate policy: lessons from tobacco control. PMID- 20006120 TI - The right oral antithrombotics in acute coronary syndromes. PMID- 20006122 TI - Stephen Luby: promoting the importance of water for health. PMID- 20006123 TI - Too much benchmarking. PMID- 20006125 TI - Chronic obstructive pulmonary disease in non-smokers. PMID- 20006126 TI - Chronic obstructive pulmonary disease in non-smokers. PMID- 20006127 TI - Chronic obstructive pulmonary disease in non-smokers. PMID- 20006129 TI - Removing user fees? Engage the health workforce. PMID- 20006130 TI - Risk of bleeding in patients with acute myocardial infarction treated with different combinations of aspirin, clopidogrel, and vitamin K antagonists in Denmark: a retrospective analysis of nationwide registry data. AB - BACKGROUND: Combinations of aspirin, clopidogrel, and vitamin K antagonists are widely used in patients after myocardial infarction. However, data for the safety of combinations are sparse. We examined the risk of hospital admission for bleeding associated with different antithrombotic regimens. METHODS: By use of nationwide registers from Denmark, we identified 40 812 patients aged 30 years or older who had been admitted to hospital with first-time myocardial infarction between 2000 and 2005. Claimed prescriptions starting at hospital discharge were used to determine the regimen prescribed according to the following groups: monotherapy with aspirin, clopidogrel, or vitamin K antagonist; dual therapy with aspirin plus clopidogrel, aspirin plus vitamin K antagonist, or clopidogrel plus vitamin K antagonist; or triple therapy including all three drugs. Risk of hospital admission for bleeding, recurrent myocardial infarction, and death were assessed by Cox proportional hazards models with the drug exposure groups as time varying covariates. FINDINGS: During a mean follow-up of 476.5 days (SD 142.0), 1891 (4.6%) patients were admitted to hospital with bleeding. The yearly incidence of bleeding was 2.6% for the aspirin group, 4.6% for clopidogrel, 4.3% for vitamin K antagonist, 3.7% for aspirin plus clopidogrel, 5.1% for aspirin plus vitamin K antagonist, 12.3% for clopidogrel plus vitamin K antagonist, and 12.0% for triple therapy. With aspirin as reference, adjusted hazard ratios for bleeding were 1.33 (95% CI 1.11-1.59) for clopidogrel, 1.23 (0.94-1.61) for vitamin K antagonist, 1.47 (1.28-1.69) for aspirin plus clopidogrel, 1.84 (1.51 2.23) for aspirin plus vitamin K antagonist, 3.52 (2.42-5.11) for clopidogrel plus vitamin K antagonist, and 4.05 (3.08-5.33) for triple therapy. Numbers needed to harm were 81.2 for aspirin plus clopidogrel, 45.4 for aspirin plus vitamin K antagonist, 15.2 for clopidogrel plus vitamin K antagonist, and 12.5 for triple therapy. 702 (37.9%) of 1852 patients with non-fatal bleeding had recurrent myocardial infarction or died during the study period compared with 7178 (18.4%) of 38 960 patients without non-fatal bleeding (HR 3.00, 2.75-3.27, p<0.0001). INTERPRETATION: In patients with myocardial infarction, risk of hospital admission for bleeding increased with the number of antithrombotic drugs used. Treatment with triple therapy or dual therapy with clopidogrel plus vitamin K antagonist should be prescribed only after thorough individual risk assessment. FUNDING: Danish Heart Foundation and the Danish Medical Research Council. PMID- 20006131 TI - Fulminant colitis and toxic myocarditis: a unifying cause? PMID- 20006132 TI - Novel vaccines: entrepreneurial contributions and scientific challenges. PMID- 20006133 TI - A challenge to vaccinology: living organisms trap information. AB - Life couples reproduction of the cell machinery with replication of the genetic program. Both processes are linked to the expression of some information. Over time, reproduction can enhance the information of the machine. We show that accumulation of valuable information results from degradative processes required to make room for novel entities. Degradation systems act as Maxwell's demons, using energy not to make room per se, but to prevent degradation of what has some functional features. This myopic process will accumulate information, whatever its source, in a ratchet-like manner. The consequence is that genes acquired by horizontal transfer as well as viruses will tend to perpetuate in niches where they are functional, creating recurrent conditions for emergence of diseases. PMID- 20006134 TI - Type I interferons as mediators of immune adjuvants for T- and B cell-dependent acquired immunity. AB - Originally identified as antiviral substances produced by infected cells, type I interferons (IFN-I) are now known to have a wide range of additional activities within both the innate and adaptive immune response. Here we review properties of IFN-I contributing to their 'natural immune adjuvant' character, and their important role for the function of complete Freund's adjuvant (CFA) and the TLR9 dependent immune adjuvant IC31. We show data to demonstrate that treatment with IFN-I boosts the ability of vaccine/adjuvant combinations to induce peptide specific CTL in both young and old mice. We view these findings in the perspective of previous clinical applications of IFN-I for vaccination. PMID- 20006135 TI - Definition of epitopes and antigens recognized by vaccinia specific immune responses: their conservation in variola virus sequences, and use as a model system to study complex pathogens. AB - In the last few years, a wealth of information has become available relating to the targets of vaccinia virus (VACV)-specific CD4(+) T cell, CD8(+) T cell and antibody responses. Due to the large size of its genome, encoding more than 200 different proteins, VACV represents a useful model system to study immunity to complex pathogens. Our data demonstrate that both cellular and humoral responses target a large number of antigens and epitopes. This broad spectrum of targets is detected in both mice and humans. CD4(+) T cell responses target late and structural antigens, while CD8(+) T cells preferentially recognize early antigens. While both CD4(+) and CD8(+) T cell responses target different types of antigens, the antigens recognized by T(H) cells are highly correlated with those recognized by antibody responses. We further show that protein abundance and antibody recognition can be used to predict antigens recognized by CD4(+) T cell responses, while early expression at the mRNA level predicts antigens targeted by CD8(+) T cells. Finally, we find that the vast majority of VACV epitopes are conserved in variola virus (VARV), thus suggesting that the epitopes defined herein also have relevance for the efficacy of VACV as a smallpox vaccine. PMID- 20006136 TI - The epidemiology of pneumococcal infections--the Swedish experience. AB - Pneumococcal infections are major contributors to morbidity and mortality world wide and pose a major public health problem. Despite being a devastating pathogen pneumococci are common colonizers of the upper respiratory tract of healthy children. There is a need for more knowledge on the molecular epidemiology, and pathogenesis of pneumococcal infections to be able to find better strategies for prevention and treatment of these common infections. Here we discuss trends in the vaccine era of the epidemiology of pneumococcal carriage, invasive disease and antibiotic resistance development as well as present national epidemiology data from Sweden of invasive pneumococcal infections during 1987-2006. PMID- 20006137 TI - The need for new vaccines. AB - Advances in biotechnology and immunology are yielding exciting progress in the development of new biologics and vaccines. Yet in both the developed and developing world, we see a backlog of new vaccines that are licensed but not yet used, an "innovation pile-up", which may prevent individuals and societies from benefiting from protection against preventable infectious diseases. What is the "need for new vaccines"? Reviewing the vaccines environment and the place of vaccination in public health, we present our business model that we use to sustainably deliver the benefits of vaccination and review potential solutions to accelerating the introduction and adoption of under-utilised and future vaccines. PMID- 20006138 TI - Anti-infective antibodies--reviving an old paradigm. AB - Antibody therapy for infections has a long history and the development of monoclonal antibody technologies, and of increasingly ingenious techniques for making these products, has brought about a major revival in interest. In this paper the field is reviewed with particular emphasis on two topics: the role that antibodies may have in combating pandemic flu; and the prospects for giving, as a food, antibodies derived from the ovalbumin of transgenic chicken eggs, as a prophylactic against diarrhoeal disease. PMID- 20006140 TI - Recent advances in the study of human antibody responses to influenza virus using optimized human hybridoma approaches. AB - Influenza viruses exhibit a fascinating level of antigenic heterogeneity that facilitates re-infection in the human population. The human antibody repertoire also manifests endless capability for variation in the genes that specify the portion of antibody molecules interacting with epitopes. A recent explosion of techniques for isolating human monoclonal antibodies to viruses has led to isolation of new antibodies that allow glimpses into the molecular basis for recognition and escape that underlies the constant antigenic drift in influenza surface proteins. These studies also reveal evidence for lifelong persistence of immunity to some influenza viruses. PMID- 20006139 TI - Monoclonal antibody-based therapies for microbial diseases. AB - The monoclonal antibody (mAb) revolution that currently provides many new options for the treatment of neoplastic and inflammatory diseases has largely bypassed the field of infectious diseases. Only one mAb is licensed for use against an infectious disease, although there are many in various stages of development. This situation is peculiar given that serum therapy was one of the first effective treatments for microbial diseases and that specific antibodies have numerous antimicrobial properties. The underdevelopment and underutilization of mAb therapies for microbial diseases has various complex explanations that include the current availability of antimicrobial drugs, small markets, high costs and microbial antigenic variation. However, there are signs that the climate for mAb therapeutics in infectious diseases is changing given increasing antibiotic drug resistance, the emergence of new pathogenic microbes for which no therapy is available, and development of mAb cocktail formulations. Currently, the major hurdle for the widespread introduction of mAb therapies for microbial diseases is economic, given the high costs of immunoglobulin preparations and relatively small markets. Despite these obstacles there are numerous opportunities for mAb development against microbial diseases and the development of radioimmunotherapy provides new options for enhancing the magic bullet. Hence, there is cautious optimism that the years ahead will see more mAbs in clinical use against microbial diseases. PMID- 20006141 TI - Prostate cancer immunotherapy yields superior long-term survival in TRAMP mice when administered at an early stage of carcinogenesis prior to the establishment of tumor-associated immunosuppression at later stages. AB - Prostate cancer immunotherapy clinical trials have been performed, but often in immunocompromised patients with limited clinical success. The study aim was to determine whether the stage of prostate cancer development at which immunization occurs affects vaccine efficacy, and if so which tumor-associated immunosuppressive mechanisms may be involved at later stages. Therapeutic vaccination of TRAMP mice with only precancerous PIN lesions confered superior protection to immunization after development of invasive carcinoma. The presence of Treg, upregulation of tumor indoleamine-2,3-dioxygenase and TGFbeta and an immunosuppressive intratumoral cytokine milieu were identified in more advanced prostate cancer. These results indicate that prostate cancer immunotherapy trials will be more successful if conducted in patients with less advanced disease. PMID- 20006142 TI - Safety and immunogenicity of an influenza vaccine A/H5N1 (A/Vietnam/1194/2004) when coadministered with a heat-labile enterotoxin (LT) adjuvant patch. AB - BACKGROUND: The use of adjuvants to enhance the immune response to novel pandemic influenza vaccine candidates may overcome the poor immune responses seen in immunologically naive populations. The confluence of a highly pathogenic H5N1 influenza virus and the widespread absence of pre-existing immunity has driven the search for effective strategies for immunization in the face of a lethal pandemic. The potent adjuvant, heat labile enterotoxin from E. coli (LT), placed over the immunization site in a patch, is a novel adjuvant strategy for immune enhancement, and was evaluated using an H5N1 injectable vaccine. METHODS: In this observer-blind, placebo-controlled clinical study, 500 healthy adults 18-49 years of age were randomized to receive two intramuscular doses of A/Vietnam/1194/2004 A/H5N1 vaccine (5microg, 15microg or 45microg) or placebo (saline) 21 days apart. For each of the influenza vaccine doses, a 50microg LT adjuvant patch was applied over the injection site at either the second or both immunizations and the HI responses (titers) were compared to H5N1 vaccine alone. The study's primary endpoint was safety, and secondary immunogenicity endpoints were evaluated using European (CHMP) licensure criteria. RESULTS: The vaccine was safe and well tolerated, and subjects generally lacked pre-existing H5N1 immunity. The single dose injection 45microg HA/LT patch regimen met all CHMP licensure criteria, including a 73% seroprotection rate compared to 49% seroprotection without a patch. Significant adjuvant effects were seen at all HA doses on Day 21. By contrast, only modest adjuvant effects were observed with the boosting regimen in subjects first primed with H5N1 alone and given the adjuvant patch only on the second immunization. The two-injection/two-patch 45microg HA regimen achieved significantly higher titers and GMFR compared to injection alone (GMFR 33.1 vs. 16.9, HI 226 vs. 94, p<0.05) and a 94% seroprotection rate. CONCLUSIONS: The LT adjuvant patch placed over the injection site was safe, significantly enhanced the immune response to an H5N1 candidate vaccine, and achieved a 73% seroprotection rate after a single dose. The LT adjuvant patch has more modest benefits in recently primed populations similar to other candidate vaccine adjuvants, but a two-dose patch plus injection regimen resulted in robust HI responses. PMID- 20006143 TI - Development of the RTS,S/AS malaria candidate vaccine. AB - A vaccine against malaria which complements existing control tools is an urgent medical need. RTS,S/AS, a pre-erythrocytic candidate vaccine, which targets the circumsporozoite protein, is the most advanced in clinical development. The safety, immunogenicity and efficacy of this candidate vaccine have been investigated in a series of trials in children and infants in endemic African countries. The vaccine shows promise for providing important public health benefits and a multicenter Phase III trial has started in Africa, aiming to further characterize the efficacy of the candidate vaccine and generate the regulatory data required for the licensing approval of the vaccine. PMID- 20006144 TI - Development of a rotavirus vaccine: clinical safety, immunogenicity, and efficacy of the pentavalent rotavirus vaccine, RotaTeq. AB - Initial approaches for rotavirus vaccines were based on the classical "Jennerian" approach and utilized simian and bovine rotavirus strains, which provided cross protection against human rotavirus strains but did not cause illness in infants and young children because of their species-specific tropism. The demonstrated efficacy of these vaccines was not consistent across studies. Thus, human-animal reassortants containing an animal rotavirus backbone with human rotavirus surface G and/or P proteins were developed, which demonstrated more consistent efficacy than that observed with the non-reassortant rotavirus strains. The pentavalent rotavirus vaccine, RotaTeq, contains 5 human-bovine reassortant rotaviruses consisting of a bovine (WC3) backbone with human rotavirus surface proteins representative of the most common G (G1, G2, G3, G4) or P (P1A[8]) types worldwide. The present review focuses on the development of the pentavalent rotavirus vaccine RotaTeq. Results of a large-scale Phase III clinical study showed that three doses of RotaTeq were immunogenic, efficacious, and well tolerated with no increased clinical risk of intussusception. RotaTeq was efficacious against rotavirus gastroenteritis of any severity (74%) and severe disease (98-100%), using a validated clinical scoring system. Reductions in rotavirus-associated hospitalizations and emergency department (ED) visits, for up to 2 years post-vaccination, were 95% in Europe, 97% in the United States, and 90% in the Latin American/Caribbean regions. RotaTeq was recently shown to be up to 100% effective in routine use in the US in reducing hospitalizations and ED visits and 96% effective in reducing physician visits. Additional studies in 8 different locations in the US have shown 85-95% reduction in rotavirus-associated hospitalizations and/or ED visits in the first 2-2.5 years of routine use. PMID- 20006145 TI - A risk-benefit analysis of vaccination. AB - Impressive success has been achieved worldwide with population based immunisation programmes against several serious infectious diseases. This success is being threatened: the incidence of previously frequent, potentially devastating diseases decreases as a consequence of successful immunisation programmes and attention of the public shifts towards true and alleged "side effects" of vaccines. This can lead to the dilemma of waning public confidence in the necessity, tolerability and safety of vaccinations. Therefore, continuous evaluation of risks and benefits of vaccines is required and results of these need to be communicated broadly to strengthen the confidence in immunisation programmes. PMID- 20006146 TI - What works? What does it cost? Comparative studies will drive decisions in health care reform. PMID- 20006147 TI - Characterization of craniomaxillofacial battle injuries sustained by United States service members in the current conflicts of Iraq and Afghanistan. AB - PURPOSE: To characterize and describe the craniomaxillofacial (CMF) battlefield injuries sustained by US Service Members in Operation Iraqi Freedom and Operation Enduring Freedom. PATIENTS AND METHODS: The Joint Theater Trauma Registry was queried from October 19, 2001, to December 11, 2007, for CMF battlefield injuries. The CMF injuries were identified using the "International Classification of Diseases, Ninth Revision, Clinical Modification" codes and the data compiled for battlefield injury service members. Nonbattlefield injuries, killed in action, and return to duty cases were excluded. RESULTS: CMF battlefield injuries were found in 2,014 of the 7,770 battlefield-injured US service members. In the 2,014 injured service members were 4,783 CMF injuries (2.4 injuries per soldier). The incidence of CMF battlefield injuries by branch of service was Army, 72%; Marines, 24%; Navy, 2%; and Air Force, 1%. The incidence of penetrating soft-tissue injuries and fractures was 58% and 27%, respectively. Of the fractures, 76% were open. The location of the facial fractures was the mandible in 36%, maxilla/zygoma in 19%, nasal in 14%, and orbit in 11%. The remaining 20% were not otherwise specified. The primary mechanism of injury involved explosive devices (84%). CONCLUSIONS: Of the injured US service members, 26% had injuries to the CMF region in the Operation Iraqi Freedom/Operation Enduring Freedom conflicts during a 6-year period. Multiple penetrating soft-tissue injuries and fractures caused by explosive devices were frequently seen. Increased survivability because of body armor, advanced battlefield medicine, and the increased use of explosive devices is probably related to the elevated incidence of CMF battlefield injuries. The current use of "International Classification of Diseases, Ninth Revision, Clinical Modification" codes with the Joint Theater Trauma Registry failed to characterize the severity of facial wounds. PMID- 20006148 TI - Endoscope-assisted rigid fixation for intraoral vertical subsigmoid osteotomy: a preliminary clinical study. AB - PURPOSE: To illustrate the clinical technique of endoscope-assisted rigid fixation in intraoral vertical subsigmoid osteotomy and to report on early postoperative morbidities. MATERIALS AND METHODS: Six patients presenting with Class III skeletal profile were recruited. The osteotomy was performed through an intraoral route. Rigid fixation was achieved with a 3-mm stab incision located inferior to the ear pinna, allowing access to the transbuccal trocar. A rigid endoscope was introduced intraorally to improve visibility during fixation. Each patient's preoperative and 3-month postoperative radiographs and clinical morbidities (neurosensory status and temporomandibular joint function) were assessed. RESULTS: Most patients (83.3%) fully recovered inferior alveolar nerve function, and 66.6% recovered temporomandibular joint function. The scar from the stab incision was effectively camouflaged by the ear pinna and was not noticeable to the patients. CONCLUSION: This preliminary study confirms that the application of endoscope-assisted rigid fixation in intraoral vertical subsigmoid osteotomy is clinically feasible. All patients presented with minimal clinical morbidities and good stability at the early postoperative period. PMID- 20006149 TI - Rocuronium and vecuronium do not affect mandibular bone marrow and masseter muscular blood flow in rabbits. AB - PURPOSE: The goal of this study was to investigate the effect of rocuronium and vecuronium continuous infusion on oral tissue blood flow in rabbits. MATERIALS AND METHODS: We used 8 male Japan White rabbits. The infusion rates of rocuronium were 7, 14, and 28 microg kg(-1) min(-1) for 20 minutes, in this order. After rocuronium was discontinued and body movement confirmed, continuous infusion of vecuronium was started. The infusion rates of vecuronium were 1.6, 3.2, and 6.4 microg kg(-1) min(-1) for 20 minutes, in this order. Observed variables were systolic blood pressure, diastolic blood pressure, mean arterial pressure, heart rate, common carotid artery blood flow, tongue mucosal blood flow, oxygen partial pressure of the mandibular bone marrow, and oxygen partial pressure of the masseter muscle. RESULTS: Heart rate in both groups tended to decrease depending on the infusion rate. Common carotid artery blood flow in the rocuronium group was increased depending on the infusion rate. Tongue mucosal blood flow in the vecuronium group was decreased depending on the infusion rate. There were no differences in diastolic blood pressure, mean arterial pressure, oxygen partial pressure of the mandibular bone marrow, and oxygen partial pressure of the masseter muscle between the 2 groups. Systolic blood pressure in both groups showed no major change. CONCLUSION: Rocuronium and vecuronium did not change mandibular bone marrow and masseter muscular blood flows. Vecuronium decreased tongue mucosal blood flow depending on the infusion rate. PMID- 20006150 TI - 18F-FDG PET in squamous cell carcinoma of the oral cavity and oropharynx: a study on inter- and intraobserver agreement. AB - PURPOSE: Good observer agreement is mandatory for an effective imaging technique. However, little is known about the observer agreement of fluorine-18 fluorodeoxyglucose ((18)F-FDG) positron emission tomography (PET) in head and neck squamous cell carcinoma. The aim of the present study was to evaluate the inter- and intraobserver agreement of interpretations of (18)F-FDG PET in head and neck SCC and to assess the influence of observer experience, tumor localizing, and tumor size on the agreement. PATIENTS AND METHODS: (18)F-FDG PET scans of 80 patients with oral and oropharyngeal SCC were reassessed twice by 2 experienced nuclear medicine physicians and 2 residents in nuclear medicine. The absolute agreement and Cohen's kappa were calculated by comparing the results of the 4 observers for the primary tumor, cervical metastases, and distant metastases/second primary tumor. To analyze the sensitivity and specificity, the results were compared with the findings from the histologic specimens or the follow-up data. RESULTS: The interobserver agreement of the nuclear medicine physicians revealed an absolute agreement and kappa of 0.91 and 0.58 for detecting the primary tumor, 0.94 and 0.83 for detecting cervical metastases, and 0.85 and 0.53 for detecting distant metastases/second primary tumors, respectively. The intraobserver agreement was greater overall than the interobserver agreement. Compared with the nuclear medicine physicians, the residents scored lower in interobserver agreement. The interobserver agreement decreased when localizing the malignancy more precisely. The agreement and sensitivity increased with tumor size. However, for small metastases, a high observer agreement was found owing to the nondetection of these malignancies. CONCLUSIONS: Good inter- and intraobserver agreement in SCC in the oral cavity or oropharynx with (18)F-FDG PET was found. Observer experience had limited influence on observer agreement. However, the agreement level decreased when a more precise anatomic tumor localization was required. PMID- 20006151 TI - Navigation-guided reduction and orbital floor reconstruction in the treatment of zygomatic-orbital-maxillary complex fractures. AB - PURPOSE: To evaluate the effectiveness of image-guided navigation on open reduction and orbital floor reconstruction as treatment for zygomatic-orbital maxillary complex fractures. PATIENTS AND METHODS: Six patients with zygomatic orbital-maxillary complex fractures were enrolled in the present study. With preoperative planning and 3-dimensional simulation, the normal anatomic structures of the deformed area were recreated by superimposing and comparing the unaffected side with the affected side. The position of dislocated bone for reduction was defined, and surgical simulation was performed. All patients underwent open reduction and orbital floor reconstruction under the guidance of the navigation system. RESULTS: A fairly accurate match between the intraoperative anatomy and the computed tomography images was achieved through registration, with a systematic error of 1-mm difference. With guidance of the navigation system, open reduction of zygomatic-orbital-maxillary complex fractures and orbital floor reconstruction were performed in all cases. The reduction was checked by postoperative computed tomography scans, with a good match with preoperative planning noted. The maximal deviation between the reduction and preoperative planning was less than 2 mm. The symptoms associated with the orbital floor defects were eliminated, and the postoperative facial appearance of the patients was clearly improved. CONCLUSION: Navigation-guided open reduction of zygomatic-orbital-maxillary complex fractures with orbital floor reconstruction can be regarded as a valuable treatment option for this potentially complicated procedure. PMID- 20006152 TI - Influence of the upper joint surface and synovial lining in the outcome of chronic closed lock of the temporomandibular joint treated with arthroscopy. AB - PURPOSE: Temporomandibular joint (TMJ) arthroscopy has been reported to be an effective and reliable technique for the treatment of chronic closed lock (CCL) of the TMJ. The purpose of the present study was to evaluate whether the status of the joint surface and the synovial lining directly visualized with arthroscopy could determine postoperative results in patients with CCL of the TMJ. MATERIALS AND METHODS: In all, 257 of 500 patients (344 joints) fulfilled the inclusion criteria for CCL of the TMJ. Of these patients, 172 with unilateral TMJ involvement were finally selected for the study. Synovitis and chondromalacia were chosen as the main features for evaluation of the joint surface and synovial lining. Two groups of patients were established: 1) patients with scarce affectation (synovitis grades I-II and chondromalacia grades I-II); and 2) patients with severe affectation (synovitis grades III-IV and/or chondromalacia grades III-IV). Pain and maximal interincisal opening were chosen as dependent variables. All patients were assessed at 1, 3, 6, 12, and 24 months postoperatively. The paired-samples Student's t test was used to compare mean values for pain (using a visual analog scale) and maximal interincisal opening (MIO) both pre- and postoperatively. The Student's t test for unpaired data was applied for the statistical analysis. A P value less than .05 was considered statistically significant. RESULTS: Synovitis grades I-II were arthroscopically observed in 87 (50.58%) patients, whereas synovitis grades III-IV were present in 72 (41.86%) patients. Chondromalacia grades I-II were arthroscopically observed in 66 (38.37%) patients, whereas chondromalacia grades III-IV were present in 54 (31.39%) patients. A statistically significant decrease in pain (P < .001) with a parallel increase in mouth opening (P < .001) after arthroscopy was observed for patients with synovitis I-II, synovitis III-IV, chondromalacia I-II, and chondromalacia III-IV during the whole follow-up period. A significant difference (P = .01) in relation to VAS score was observed between patients with synovitis I II and patients with synovitis III-IV at month 6 postoperatively. However, this difference did not persist during the rest of the follow-up period, as was the case in relation to mouth opening. No significant differences were observed in relation to decrease of pain and increase of MIO between patients with chondromalacia I-II and patients with chondromalacia III-IV at any time during the follow-up period. Although mean values for pain were lower in patients with synovitis I-II plus chondromalacia I-II in comparison to patients with synovitis III-IV plus chondromalacia III-IV for the whole follow-up period, no statistical significant differences were observed. In relation to the increase in mouth opening, slightly higher values were observed for patients with synovitis I-II plus chondromalacia I-II, although no statistical differences were observed with regard to patients presenting with synovitis III-IV plus chondromalacia III-IV. CONCLUSION: A significant decrease in pain with a parallel increase in MIO was achieved from month 1 postoperatively in patients with any grade of synovitis and/or chondromalacia. No statistical difference in pain or function was observed between patients with scarce involvement of the joint surface and the synovial lining and patients with severe involvement after arthroscopy. PMID- 20006153 TI - Influence of oral hygiene and smoking on pain and swelling after surgical extraction of impacted mandibular third molars. AB - PURPOSE: To assess the pain and swelling during the first week after surgical extraction of impacted mandibular third molars and the relationship with oral hygiene and smoking before surgery and during the postoperative period. MATERIALS AND METHODS: A prospective study was performed on patients undergoing surgical extractions of impacted mandibular third molars. Pain was recorded on a visual analog scale from 1 to 10 and swelling on a 4-point descriptive scale at 2, 6, and 12 hours after surgery and daily during the first postoperative week. Oral hygiene and smoking before surgery and during the postoperative period were recorded. Statistical analysis was performed of all the variables. A P value less than .05 was considered statistically significant. RESULTS: A total of 50 patients (27 men and 23 women) with a mean age of 26.3 years (range 18 to 39) underwent surgical extraction of an impacted third molar. The maximum pain occurred during the first day and the maximum swelling at 24 hours after surgery. The patients with a lower brushing frequency before surgery reported greater pain. Likewise, the patients who smoked more after surgery experienced greater pain at 24 hours postoperatively. The hygiene after surgery and smoking before the surgical intervention had no statistically significant relationship to the occurrence of pain (P > .05). Swelling had no relationship with the variables studied (P > .05). CONCLUSIONS: Surgical extraction of an impacted third molar caused moderate pain and swelling during the first 24 hours after surgery. A lower brushing frequency before surgery and during the first postoperative week as well as smoking after surgery were related to greater pain scores. PMID- 20006154 TI - Unilateral condylar hyperactivity: a histopathologic analysis of 47 patients. AB - PURPOSE: To perform a histopathologic analysis of condyles that were resected because of unilateral condylar hyperactivity and compare the results of the bone scan with the histopathologic findings. PATIENTS AND METHODS: A total of 47 resected condyles were histopathologically examined. In 29 cases, a single photon emission computed tomography (SPECT) bone scan was available. For all condylar specimens, a standardized histologic scoring system was used to assess the number of cartilage islands and the thickness of the cartilage layer. The SPECT scans were analyzed by calculating the difference in bone activity between the hyperactive and contralateral condyles. RESULTS: The number of cartilage islands was highly variable, ranging from almost absent in 37% of the patients to abundant in 35%. Furthermore, the relative thickness of the cartilage layer exhibited considerable variation, from less than one quarter of the total thickness of the condylar articular layer in 22% of the patients to one half of the total thickness in 35%. We found no significant relationship between the number of cartilage islands and bone activity using SPECT (P = .11) or between the relative thickness of the cartilage layer and bone activity using SPECT (P = .82). CONCLUSIONS: Unilateral condylar bone growth can occur without large numbers of cartilage islands and without abundant cartilage formation. The bone activity measured by bone scintigraphy was not related to the histologic results. The histopathologic findings of resected condyles in unilateral condylar hyperactivity cannot, therefore, be used as a reference standard. Nevertheless, histopathologic examination should always be performed to rule out other diseases. PMID- 20006155 TI - Blood replacement practices for complex orthognathic surgery: a single surgeon's experience. AB - PURPOSE: The purpose of this study is to review the blood replacement practices in a consecutive series of a single surgeon's experience whose patients all underwent, at a minimum, simultaneous Le Fort I maxillary osteotomy, bilateral sagittal split osteotomies of the mandible, septoplasty, and inferior turbinate reduction procedures. PATIENTS AND METHODS: A consecutive series of a single surgeon's patients who met inclusion criteria of (n = 34) during a 5-month time frame were included. Records included office charts, hospital records, and data stored at the Red Cross (hospital) blood bank. RESULTS: A total of 76% (26/34) of the study patients chose to auto donate [corrected].. Only 2 of the study patients underwent blood transfusion (6%). One of the transfused patients received 1 unit of auto-donated blood, whereas the other transfused patient received a unit of homologous packed red blood cells. Based on the total units of blood predonated, 97% (28/29) of stored units were discarded. CONCLUSION: Only a small percentage (6%) of individuals undergoing complex orthognathic and intranasal surgery received blood replacement. We believe that close collaboration between the surgical and anesthesia teams and the recovery of patients in a safely monitored environment will continue to reduce the need for transfusion in the orthognathic patient. PMID- 20006156 TI - Stability behavior of human tibias after bone removal--comparative examination in 15 cadaver tibia pairs. AB - PURPOSE: To obtain scientific information on the loss of stability of tibias after removal of bone grafts, we performed a comparative study of 15 freshly preserved adult cadavers to determine the axial breaking loads of the operated and nonoperated tibial heads. MATERIALS AND METHODS: From all cadavers, 1 tibia was randomly selected from which the maximum possible amount of cancellous bone was harvested. The respective contralateral side remained untouched. After maceration, the proximal tibias of each cadaver were removed bilaterally and adjusted to precisely equal lengths. Using a Zwick universal testing machine, the tibial heads were loaded by an axial force until fracture. As the final breaking load, the force value was recorded when the first distinct decrease in the feed force curve was observed. To compare the mean breaking loads of the operated and nonoperated control tibias, a t test for related samples at P = .05 was used. RESULTS: The mean breaking load for the donor tibias was 3,767 N and was significantly lower than that of the control side with an average of 5,126 N. This finding was independent of age and gender. CONCLUSIONS: Bone removal from the proximal tibia leads to a significant reduction of the axial load capacity. Therefore, we recommend partial loading of up to one half of the body weight during the first postoperative week. For an additional 5 weeks, patients should bear their full body weight on the affected leg only when walking normally and on flat ground. PMID- 20006157 TI - Sialoendoscopically assisted open sialolithectomy for removal of large submandibular hilar calculi. AB - PURPOSE: The management of large hilar calculi is a technically challenging issue during sialoendoscopic surgery. The aim of the present study was to evaluate the clinical efficacy of sialoendoscopically assisted open sialolithectomy for the removal of large submandibular hilar calculi to avoid sialoadenectomy. PATIENTS AND METHODS: The present study was undertaken among patients with sialolithiasis scheduled for sialoendoscopic surgery from August 2005 to October 2008. When we failed to remove large submandibular hilar stones intraductally, we performed sialoendoscopically assisted open sialolithectomy. The clinical characteristics, pre- and intraoperative data, and outcomes were documented in a prospective fashion. RESULTS: Of 78 consecutive patients with submandibular sialolithiasis, 18 were treated with sialoendoscopically assisted open sialolithectomy immediately after failure of intraductal removal of calculi by sialoendoscopy. For 17 patients, large hilar sialoliths were successfully removed using this surgical technique. The surgery failed in 1 patient with multiple sialoliths, and the procedure was converted to open sialoadenectomy. Temporary numbness of the tongue for 1 week postoperatively was documented in 3 patients. The patients were followed up for a median period of 18 months without any symptoms or signs of recurrence. CONCLUSION: Our results suggest that sialoendoscopically assisted open sialolithectomy is an effective and safe surgical technique to remove large submandibular hilar calculi. PMID- 20006158 TI - Diagnostic accuracy of panoramic radiography in determining relationship between inferior alveolar nerve and mandibular third molar. AB - PURPOSE: The aim of this review was to determine the diagnostic accuracy of panoramic radiographic markers in the detection of the relationship between the mandibular canal and third molar roots. MATERIALS AND METHODS: A literature search of electronic databases, Cochrane Oral Health Group's Trials Register, National Research Register, conference proceedings, and abstracts was performed to identify studies that had investigated the diagnostic accuracy of the 3 panoramic radiographic markers (ie, darkening of the root, interruption of the radiopaque borders, and diversion of the mandibular canal). RevMan, version 5.0, and Meta-DiSc software programs were used for the pooled analyses and the construction of a summary receiver operating characteristic curve. RESULTS: A total of 5 studies were included, involving 894 observations. The overall pooled sensitivity and specificity for darkening of the root was calculated as 51.2% (95% confidence interval [CI] 42% to 60%) and 89% (95% CI 87% to 90%), respectively. The interruption of radiopaque borders showed a pooled sensitivity of 53.5% (95% CI 78.1% to 81.8%) and a pooled specificity of 80% (95% CI 78.1% to 81.8%). The diversion of the canal criterion had a pooled sensitivity of 29.4% (95% CI 21.8% to 38.1%) and a pooled specificity of 94.7% (95% CI 93.6% to 95.7%). The area under the receiver operating characteristic curve was 70% to 77%. CONCLUSIONS: The results of this meta-analysis suggest a reasonable diagnostic accuracy for panoramic radiography in the preoperative evaluation of the relationship between third molars and the canal. Additional studies are needed to examine a more accurate, accessible, and cost-effective initial radiographic technique before third molar surgery. PMID- 20006159 TI - Chronic obstructive parotitis due to strictures of Stenson's duct--our treatment experience with sialoendoscopy. AB - PURPOSE: The present retrospective analysis was performed to assess the effectiveness of sialoendoscopic treatment of chronic obstructive parotitis. MATERIALS AND METHODS: The study group included 87 parotid glands with stricture as the only cause of gland obstruction. After sialographic and sialoendoscopic evaluation, sialoendoscopic 4-step surgical treatments were performed. The treatment protocol was designed. RESULTS: Sialoendoscopy proved to be a successful procedure in 94.7% of the affected parotid glands we treated. CONCLUSION: The minimally invasive technique of sialoendoscopy for treatment of strictures of the salivary glands could be a welcome innovation, helping to avoid radical surgical treatment of salivary gland disease. PMID- 20006160 TI - Time used for orthodontic surgical treatment of dentofacial deformities in white patients. AB - PURPOSE: The aim of the present study was to assess the duration of each treatment phase that a patient with dentofacial disorders and eligible for orthodontic surgery undergoes and to identify those factors that determine the time. We have also included a description of the demographic and skeletal characteristics of the patients treated at our center, the surgery type, and movements performed. PATIENTS AND METHODS: A sample of 148 patients undergoing orthognathic surgery from 2004 to 2007 at Princesa University Hospital, Madrid, was studied. The demographic data of each patient, their facial pattern, skeletal disorders, and surgical movements performed were recorded, as was the duration of orthodontic treatment, both pre- and postoperatively, length of surgery, and length of hospital stay. RESULTS: The length of preoperative orthodontic treatment was 24 months and that of postoperative treatment was 12 months. The average operative time was 4 hours for those interventions in which both dental arches were involved, and the average length of hospital stay was 3 days. No relevant differences were found for the length of orthodontic treatment according to gender, age, or type of skeletal disorder. Differences were observed in the length of hospital stay between those patients who underwent segmentation of the maxilla and those who did not. CONCLUSIONS: The length of the different phases into which an orthodontic surgical treatment is divided can differ within some given limits. However, it does not depend on either the patients' demographic skeletal characteristics or the surgical movements performed. PMID- 20006161 TI - Maxillofacial air-containing cavities, blast implosion injuries, and management. AB - PURPOSE: Distinctive mechanisms of primary blast effects have produced a transitional era of facial trauma. Implosion mechanism damage is one of these. Implosion mechanism damage results in injury limited to the gas-containing structures of the auditory canal, paranasal sinuses, gastrointestinal tract, and lungs. Worldwide, the victims of explosive detonations have increased and advanced dramatically. The outcome is greater mortality and morbidity and new types of injuries, especially in the maxillofacial region. Thus, the knowledge of, and experience with, their management should be shared globally by colleagues through publications. MATERIALS AND METHODS: The implosion and mini re-explosion of compressed air sinuses leads to skeletal crush injury to the nasal-orbital ethmoidal, maxillary sinuses, and nasal bones. A variety of surgical approaches were used successfully under conditions of war. The assessment of the associated injuries to the lung and/or brain is the initial priority to any life-threatening blast injury. This article describes the biophysical results of blast injuries to the middle third facial skeleton and associated injuries and details the management and protection of crushed air containing paranasal spaces. RESULTS: Easy, simple, and fast treatment and management were used successfully on the pulverized, fragmented skeletal architecture of the facial middle part without increasing morbidity and with the avoidance of unnecessary surgical trauma. CONCLUSIONS: Injuries in one of the most difficult esthetic, physiologic, and anatomic regions of the body is best treated with an understanding of the biophysical effects of the implosion mechanism on air-containing spaces in the maxillofacial region. The introduction of new methods for the management of severe destruction of hard and soft tissue will decrease the incidence of complications and the operative time. PMID- 20006162 TI - Quality-of-life survey comparing patients before and after discectomy of the temporomandibular joint. AB - PURPOSE: The aim of this study was to assess the quality-of-life (QoL) outcomes of patients who had undergone temporomandibular joint (TMJ) discectomy with dermis-fat grafting compared with a cohort of closely matched patients who had not had surgery. MATERIALS AND METHODS: A cross-sectional study of 61 patients was undertaken. All patients completed a TMJ surgery-specific QoL questionnaire. They were divided into 2 groups according to whether they had undergone TMJ discectomy (postsurgical group, n = 32) or not (presurgical group, n = 29). The 2 groups were closely matched for age, gender, clinical presentation, and radiologic diagnoses of Wilkes stage IV TMJ internal derangement. RESULTS: Post TMJ surgery patients showed statistically significant decreases in pain levels (P < .05), diet and chewing (P < .01), mood (P < .01), anxiety (P < .01), and general health (P < .05) compared with the presurgical patients. However, there were no statistically significant differences between the pre- and post-TMJ surgery groups in terms of speech, level of activity, recreation, and general well-being. CONCLUSION: The results of this study suggest that TMJ discectomy with dermis-fat grafting appears to have a positive QoL effect in terms of reducing pain levels and improving diet and chewing, mood, anxiety, and general health in patients with Wilkes stage IV TMJ internal derangement. PMID- 20006163 TI - Tooth extraction in patients taking intravenous bisphosphonates: a preventive protocol and case series. AB - PURPOSE: To test the efficacy of a protocol in preventing the development of bisphosphonate-related osteonecrosis of the jaw (BRONJ) after tooth extraction. PATIENTS AND METHODS: In this prospective case series, consecutive subjects treated with intravenous bisphosphonates who needed tooth extraction underwent a protocol aimed at reducing the risk of BRONJ, based on local and systemic infection control by means of mechanical and chemical reduction of the local bacterial load plus antibiotic prophylaxis. RESULTS: We performed 38 extractions in 23 patients treated with intravenous bisphosphonates, mainly zoledronate, for a mean of 17.5 months (range, 3-36 months). Five patients already had signs of BRONJ caused by tooth extractions performed elsewhere. The mean follow-up was 229.5 days (range, 14-965 days), and no case of BRONJ was recorded. CONCLUSIONS: Despite the methodologic limitations of the study design, the proposed preventive protocol appears to reduce the risk of BRONJ after tooth extraction in a group of subjects treated with intravenous bisphosphonates. PMID- 20006164 TI - Tooth replantation after use of Euro-Collins solution or bovine milk as storage medium: a histomorphometric analysis in dogs. AB - PURPOSE: Euro-Collins solution was developed for the preservation of organs for transplantation, whose characteristics have raised interest for its use as a storage medium for avulsed teeth before replantation. This study evaluated histologically and morphometrically the healing process of dog teeth replanted after storage in Euro-Collins solution or bovine milk. MATERIALS AND METHODS: Eighty roots of 4 young adult mongrel dogs were randomly assigned to 4 groups (n = 20) and the root canals were instrumented and obturated with gutta-percha and a calcium hydroxide-based sealer. After 2 weeks, the teeth were extracted and subjected to the following protocols: GI (negative control), replantation immediately after extraction; GII (positive control), bench-drying for 2 hours before replantation; GIII and GIV, immersion in 10 mL of whole bovine milk and Euro-Collins solution at 4 degrees C, respectively, for 8 hours before replantation. The animals were sacrificed 90 days postoperatively. The pieces containing the replanted teeth were subjected to routine processing for histologic and histometric analyses under light microscopy and polarized light microscopy. RESULTS: Root resorption was observed in all groups. GII exhibited the greatest loss of dental structure (P < .01), and inflammatory resorption was predominant in this group. Storage in milk showed poorer results than immediate replantation and storage in Euro-Collins solution (P < .01). The teeth stored in Euro-Collins solution presented similar extension of root resorption and periodontal ligament reorganization to those of immediately replanted teeth. CONCLUSIONS: The findings of this study suggest that the Euro-Collins solution is an adequate storage medium for keeping avulsed teeth for up to 8 hours before replantation. PMID- 20006165 TI - Efficacy and safety of oral propranolol premedication to reduce reflex tachycardia during hypotensive anesthesia with sodium nitroprusside in orthognathic surgery: a double-blind randomized clinical trial. AB - PURPOSE: The present study sought to determine whether premedication with oral propranolol 10 mg before hypotensive anesthesia with sodium nitroprusside could reduce reflex tachycardia, the amount of sodium nitroprusside used, and blood loss during hypotensive anesthesia for orthognathic surgery. PATIENTS AND METHODS: A total of 60 patients undergoing bimaxillary surgery were studied in a prospective, randomized, and double-blind study of oral propranolol 10 mg or placebo as premedication before hypotensive anesthesia with sodium nitroprusside. Hemodynamic variables, the amount of sodium nitroprusside used, and blood loss were statistically analyzed. RESULTS: The heart rate and amount of sodium nitroprusside used were highly significantly less (P < .01) in the propranolol group, but no significant difference was found in blood loss between the 2 groups. No clinically significant complications were observed in either group. CONCLUSION: Premedication with oral propranolol 10 mg before hypotensive anesthesia with sodium nitroprusside is safe and effective to reduce reflex tachycardia and the amount of sodium nitroprusside used. PMID- 20006166 TI - Tetracycline bone fluorescence: a valuable marker for osteonecrosis characterization and therapy. AB - PURPOSE: Differential diagnosis of osteoradionecrosis and bisphosphonate-related osteonecrosis of the jaw is primarily based on medical history, rather than pathogenesis or histopathology. This report aims to redress this shortcoming by demonstrating the advantages of tetracycline bone fluorescence as an aid to characterize osteonecrosis entities according to differential histopathologies. Furthermore, this technique facilitates the means to determine extent of necrosis and to optimize surgical therapy. PATIENTS AND METHODS: Two patients with extended osteonecrosis of the lower jaw (osteoradionecrosis or bisphosphonate related osteonecrosis of the jaw) were treated with partial mandibulectomy. After preoperative administration of doxycycline for 10 days, bone fluorescence was monitored intraoperatively to determine the resection boundaries. RESULTS: Fluorescence analysis correlated well with the specific histopathologic features of the 2 osteonecrosis entities. Bone fluorescence was predominantly observed in the cortical bone and cancellous bone regions in osteoradionecrosis and bisphosphonate-related osteonecrosis of the jaw, respectively. Margins of the osteonecrosis (and the resection) could be determined under fluorescence guidance; however, bone bleeding did not correlate with bone fluorescence in both osteonecrosis entities. CONCLUSIONS: Given that viable but not necrotic bone displays tetracycline fluorescence, a notion that reflects the histopathology, more precise characterization of the 2 osteonecrosis types is enabled. Furthermore, even in extended cases of osteonecrosis requiring partial mandibulectomy, bone fluorescence helps to pinpoint the margins of resection and thus signifies an improvement of surgical therapy of extended osteonecrosis. PMID- 20006168 TI - Intranasal desmopressin versus blood transfusion in cirrhotic patients with coagulopathy undergoing dental extraction: a randomized controlled trial. AB - PURPOSE: Cirrhotic patients waiting for liver transplantation who need dental extractions are given fresh frozen plasma and/or platelets to correct coagulopathy. This is costly and may be associated with transfusion reactions and fluid overload. We evaluated the efficacy of intranasal desmopressin as an alternative to transfusion to correct the coagulopathy of cirrhotic patients undergoing dental extraction. PATIENTS AND METHODS: Cirrhotic patients with platelet counts of 30,000 to 50,000/microL and/or international normalized ratio (INR) 2.0 to 3.0 were enrolled in a prospective, controlled, randomized clinical trial. Blood transfusion (fresh frozen plasma 10 mL/kg and/or 1 unit of single donor platelets, respectively) or intranasal desmopressin (300 microg) were given before dental extraction. A standard oral and maxillofacial surgical treatment protocol was performed by the same surgeon. Patients were followed for postextraction bleeding and side-effects over the next 24 to 48 hours. RESULTS: No significant differences were noted between the 2 groups in gender, age, INR, platelet count, creatinine, total bilirubin, ALT, albumin, MELD score, or number of teeth removed (median 3 vs 4). The number of teeth removed ranged between 1 and 31 in the desmopressin group and 1 and 22 in the transfusion group. No patients in desmopressin group required rescue blood transfusion after extraction. One patient in the transfusion group had bleeding after the procedure and required an additional transfusion. Another patient experienced an allergic reaction at the end of transfusion, which was effectively treated with diphenhydramine. Treatment associated average costs were lower for desmopressin ($700/patient) compared with transfusion ($1,173/patient). CONCLUSIONS: Intranasal desmopressin was as effective as blood transfusion in achieving hemostasis in cirrhotic patients with moderate coagulopathy undergoing dental extraction. Intranasal desmopressin was much more convenient, less expensive, and well tolerated. PMID- 20006167 TI - Maxillofacial and axial/appendicular giant cell lesions: unique tumors or variants of the same disease?--A comparison of phenotypic, clinical, and radiographic characteristics. AB - PURPOSE: The relationship between giant cell lesions (GCLs) of the maxillofacial (MF) skeleton and those of the axial/appendicular (AA) skeleton has been long debated. The present study compared the clinical and radiographic characteristics of subjects with MF and AA GCLs. MATERIALS AND METHODS: This was a retrospective cohort study of patients treated for GCLs at Massachusetts General Hospital from 1993 to 2008. The predictor variables included tumor location (MF or AA) and clinical behavior (aggressive or nonaggressive). The outcome variables included demographic, clinical, and radiographic parameters, treatments, and outcomes. Descriptive and bivariate statistics were computed, and P 96 hours. As a secondary outcome, we recorded IV complications, including central line placement. We used descriptive statistics, univariate survival analysis with Kaplan Meier, and log-rank tests for data analysis. RESULTS: Seventy-five patients were enrolled. The average age was 52 years. Fifty-three percent were male, 21% obese, and 13% had a history of injection drug use. The overall IV survival rate was 56% (95% confidence interval, 44%-67%) with a median survival of 26 hours (interquartile range [IQR], 8-61). Forty-seven percent of IVs failed within 24 hours, most commonly due to infiltration. Although 47 (63%) operators reported that a central line would have been required if peripheral access was unobtainable, only 5 (7%; 95% confidence interval, 2%-15%) patients underwent central venous catheterization. Only 1 central line was placed as a result of ultrasonography-guided IV failure. We observed no infectious or thrombotic complications. CONCLUSION: Despite a high premature failure rate, ultrasonography-guided peripheral IVs appear to be an effective alternative to central line placement in ED patients with difficult access. PMID- 20006194 TI - Oral N-acetylcysteine has a deleterious effect in acute iron intoxication in rats. AB - Acute iron intoxication is associated with depletion of reduced glutathione in hepatocytes and changes in the glutathione system enzymes. We hypothesized that treatment with N-acetylcysteine (NAC), a glutathione reducing agent and an antioxidant, would reduce mortality in acute iron intoxication. We used a rat model to test this hypothesis. Male rats were assigned to 4 groups. Group 1 received 400 mg/kg elemental iron by oral gavage, group 2 received the same dose of iron followed by NAC, group 3 received NAC only, whereas group 4 received distilled water. Iron and liver transaminases in the blood, and glutathione system enzymes in the liver and erythrocytes were measured. Mortality in group 2 was significantly higher after 2, 6, and 24 hours compared with group 1 (P < .001). No deaths were observed in groups 3 and 4. Serum iron levels were significantly higher in group 2 rats compared to group 1 rats (P < .001). Hepatic and erythrocyte glutathione system enzymes were significantly lower among rats in group 2 compared to rats in group 1. The administration of NAC probably increased the absorption of iron through the gastrointestinal tract, causing higher serum iron levels with significant hepatic damage. These results indicate that in a rat model of acute iron intoxication, orally administered NAC may increase mortality. PMID- 20006195 TI - Diagnostic utility of the genital Gram stain in ED patients. AB - OBJECTIVE: The study aimed to determine the diagnostic usefulness of the genital Gram stain in an emergency department (ED) population. METHODS: A linked-query of an urban, tertiary-care, university- affiliated hospital laboratory database was conducted for all completed Chlamydia trachomatis and Neisseria gonorrhoeae DNA probes, Trichomonas vaginalis wet preps, and genital Gram stains performed on ED patient visits between January and December 2004. Positive criteria for a Gram stain included greater than 10 white blood cells per high-power field, gram negative intracellular/extracellular diplococci (suggesting N gonorrhoeae), clue cells (suggesting T vaginalis), or direct visualization of T vaginalis organisms. DNA probes were used as the gold standard definition for N gonorrhoeae and C trachomatis infection. RESULTS: Of 1511 initially eligible ED visits, 941 were analyzed (genital Gram stain and DNA probe results both present), with a prevalence of either C trachomatis or N gonorrhoeae of 11.4%. A positive genital Gram stain was 75.7% sensitive and 43.3% specific in diagnosing either C trachomatis and/or N gonorrhoeae infection, and 80.4% sensitive and 32.2% specific when the positive cutoff was lowered to more than 5 white blood cells/high-power field. No Gram stains were positive for T vaginalis (with 47 positive wet mounts), and clue cells were noted on 117 Gram stains (11.6%). CONCLUSION: Gram stains in isolation lack sufficient diagnostic ability to detect either C trachomatis or N gonorrhoeae infection in the ED. PMID- 20006196 TI - Patients with coronary disease fail observation status at higher rates than patients without coronary disease. AB - BACKGROUND: Few studies have evaluated emergency department (ED) observation unit chest pain protocols for optimal patient characteristics and admission rates. At our 35 000-visits/y ED, we implemented a chest pain protocol for our observation unit that allowed emergency physicians to admit patients with known coronary artery disease (CAD). METHODS: We performed a retrospective chart review of all observation unit patients admitted under the chest pain protocol from April 1, 2006, to May 31, 2007. We compared the outcomes of patients who had a history of CAD with those who did not. RESULTS: Five hundred thirty-one patients were admitted to the observation unit under the chest pain protocol for the 14-month study period. Of these patients, 125 (23.5%) had a history of CAD. Patients with a history of CAD had a higher inpatient admission rate ( 24% vs 8.6%; P < .001), higher rate of a positive stress test or positive coronary computed tomographic scan (32.3% vs 6.9%; P < .001), a higher rate of cardiac catheterization (12% vs 5.9%; P = .02), and a higher rate of stent placement or coronary artery bypass graft (CABG) (7.2% vs 2.2%; P = .007). In multivariate analysis, patient history of CAD was an independent predictor of hospital admission (P = .005) and stent placement or CABG (P = .030). CONCLUSION: Patients with known CAD who were admitted to the ED observation unit failed observation status (ie, required hospitalization) and had higher rates of positive testing than those without CAD. PMID- 20006197 TI - ED antibiotic use for acute respiratory illnesses since pneumonia performance measure inception. AB - OBJECTIVE: The study aimed to determine if emergency department (ED)-administered antibiotics for patients discharged home with nonpneumonia acute respiratory tract infections (ARIs) have increased since national pneumonia performance measure implementation, including antibiotic administration within 4 hours of arrival. METHODS: DESIGN: Time series analysis. SETTING: Six university and 7 Veterans Administration EDs participating in the Improving Antibiotic Use for Acute Care Treatment (IMPAACT) trial (randomized educational intervention to reduce antibiotics for bronchitis). PARTICIPANTS: Randomly selected adult (age >18 years) ED visits for acute cough, diagnosed with nonpneumonia ARIs, discharged home during winters (November-February) of 2003 to 2007. MAIN OUTCOME: Time trend in ED-administered antibiotics, adjusted for patient demographics, comorbidities, vital signs, ED length of stay, IMPAACT intervention status, geographic region, Veterans Administration/university setting, and site and provider level clustering. RESULTS: Six thousand four hundred seventy-six met study criteria. Three hundred ninety-four (6.1%) received ED-administered antibiotics. Emergency department-administered antibiotics did not increase across the study period among all IMPAACT sites (odds ratio [OR], 0.88; 95% confidence interval [CI], 0.76-1.01) after adjusting for age, congestive heart failure history, temperature higher than 100 degrees F, heart rate more than 100, blood cultures obtained, diagnoses, and ED length of stay. The ED-administered antibiotic rate decreased at IMPAACT intervention (OR, 0.80; 95% CI, 0.69-0.93) but not nonintervention sites (OR, 1.04; 95% CI, 0.91-1.19). Adjusted proportions receiving ED-administered antibiotics were 6.1% (95% CI, 2.7%-13.2%) for 2003 to 2004; 4.8% (95% CI, 2.2%-10.0%) for 2004 to 2005; 4.6% (95% CI, 2.7%-7.8%) for 2005 to 2006; and 4.2% (95% CI, 2.2%-8.0%) for 2006 to 2007. CONCLUSIONS: Emergency department-administered antibiotics did not increase for patients with acute cough discharged home with nonpneumonia ARIs since pneumonia antibiotic timing performance measure implementation in these academic EDs. PMID- 20006198 TI - How stylet use can effect confirmation of endotracheal tube position using ultrasound. AB - INTRODUCTION: None of the techniques used for confirmation of endotracheal tube (ET) placement are proven reliable 100% of the time. The purpose of our study is to determine whether ultrasound can accurately detect the passage of ET through the trachea and esophagus and to see whether this visualization is augmented with the use of a metal stylet. METHODS: A total of 7 physicians made assessments of ET positions using an ultrasound during their passage through the trachea or esophagus. A total of 40 esophageal and 40 tracheal intubations were performed randomly in a blinded fashion on a fresh, unfrozen human cadaver. Half were performed with a metal stylet and the other half without a stylet. RESULTS: During transtracheal assessment regardless of stylet use, correct identification of ET position was achieved in 275 of 280 esophageal intubations and 268 of 280 tracheal intubations. The overall sensitivity was 95.7%, and specificity was 98.2%. The presence and the absence of stylet was identified in 109 of 280 and in 155 of 280 attempts, respectively. Correct identification of stylet presence yielded a sensitivity of 38.9% and a specificity of 55.4%. Ultrasound can be used by emergency physicians to accurately detect the passage of ET through the trachea and esophagus; however, stylet use did not augment ET visualization. PMID- 20006199 TI - Can hypertonic saline influence platelet P selectin expression and platelet leukocyte aggregation? AB - OBJECTIVES: Part of platelet function involves aggregation and activation. Activation leads to platelet P selectin expression and platelet-leukocyte aggregation. Hypertonic saline inhibits platelet aggregation, although the effects of hypertonic saline on platelet activation are not known. We evaluated the effects of hypertonic saline on platelet activation as measured by platelet P selectin expression and platelet-leukocyte aggregation. METHODS: Blood samples from healthy volunteers (n = 6) were treated in vitro with various solutions including 23.5%, 7.5%, 3%, and 0.9% saline; Ringer's solution; 5% dextrose in water; and 10% hydroxyethyl starch. Blood was diluted with each type of solution to 2.5%, 5%, 10%, 20%, and 30% (vol/vol) dilution. All blood samples were activated with adenosine diphosphate (20 micromol/L), stained with fluorochrome conjugated antibodies, and analyzed by flow cytometry to measure platelet P selectin expression and platelet-leukocyte aggregation. RESULTS: The 23.5% saline solution reduced P selectin expression at 20% and 30% dilutions and platelet leukocyte aggregation at 10%, 20%, and 30% dilutions. The 7.5% solution saline had no effect on P selectin expression and significantly inhibited platelet leukocyte aggregation only at 30% dilution. Other solutions had no effect on platelet P selectin expression or platelet-leukocyte aggregation. CONCLUSIONS: Our data suggest that hypertonic saline does not affect platelet P selectin expression or platelet-leukocyte aggregation at therapeutic plasma concentrations but that an inhibitory effect occurs at supratherapeutic doses. Dilutions of other solutions caused the least disturbance of platelet activation. PMID- 20006200 TI - Validating the ABCD(2) Score for predicting stroke risk after transient ischemic attack in the ED. AB - OBJECTIVES: The aim of the study was to validate the use of the ABCD(2) score for the prediction of stroke after transient ischemic attack (TIA) in patients presenting to the emergency department (ED). The ABCD(2) scoring is based on 5 factors as follows: age of at least 60 years; blood pressure of at least 140/90 mm Hg; clinical features such as unilateral weakness and speech impairment alone; duration of at least 60 minutes or 10 to 59 minutes; and diabetes. METHODS: The authors conducted a retrospective observational study of all patients presented to the ED for TIA, as diagnosed by the attending emergency physicians, for a 2 year period. Sensitivity, specificity, and negative predictive value (NPV) were calculated for risk of stroke at 2, 7, 30, and 90 days after presentation. RESULTS: From January 1, 2005, to December 31, 2006, there were 470 patients diagnosed with TIA at the ED. Mean age was 61.0 years (SD, 13.2), with 63.3% males. Age of at least 60 years, unilateral weakness, and duration of at least 60 minutes were found to be significant predictors of stroke at 2 days. An admission rule based on an ABCD(2) score of at least 4 showed sensitivity of 86.4% and NPV of 91.7% for stroke at 7 days. Admission based on a score of at least 3 showed sensitivity of 96.6% and NPV of 96.1%. Admission rate was 69.1% and. 83.6%, respectively. CONCLUSION: The ABCD(2) rule showed good sensitivity and NPV for stroke at 7 days. However, NPV was not 100%, and there would still be patients being discharged from the ED and returning with a stroke if this cutoff was implemented in our setting. PMID- 20006201 TI - Echocardiographic evaluation of TASER X26 probe deployment into the chests of human volunteers. AB - Several animal studies have shown that the TASER X26 (TASER International, Scottsdale, Ariz) conducted electrical weapon can electrically capture the myocardium when discharged on the thorax. These results have not been reproduced in human echocardiographic studies. A primary limitation of those human studies is that the TASER device was connected by taping the wires into conductive gel on the skin surface of the thorax. This study overcomes those limitations. In this study, a training instructor discharged a TASER X26 into the chests of 10 subjects from a distance of 7 ft so that a 5-second discharge could be administered through the probes as in field exposures. Limited echocardiography was performed before, during, and after discharge. In agreement with 2 prior studies by these authors, the TASER X26 did not electrically capture the human myocardium when used with probe deployment. These data are contrary to animal studies in which capture occurred. PMID- 20006202 TI - Demographics of patient visits during high daily census in a pediatric ED. AB - PURPOSE: The aim of this study is to describe patient demographics in a pediatric emergency department (PED) during low, average, and high daily census days. METHODS: Daily PED census, between January 1, 2006, and December 31, 2006, was categorized into very low, low, average, high, and very high quintiles. Variables of interest included acuity, age, health care coverage, and disposition. chi(2) analysis assessed the significance of differences in proportions of patient populations across the census quintiles. RESULTS: An increasing proportion of younger children (<2 years of age) received care as daily volumes increased (P < .0001). Proportions of Medicaid and self-pay patient increased, whereas that of commercially insured patients decreased as daily census increased (P < .0001). The distributions of patient acuity level (63.1% nonurgent) and admission rate (12.8%) did not differ significantly cross census quintiles. CONCLUSIONS: Younger children with self-pay and government-assisted health care coverage make up a greater proportion of children seen in a PED during high census days. PMID- 20006203 TI - S-100beta and neuron-specific enolase levels in carbon monoxide-related brain injury. AB - INTRODUCTION: Carbon monoxide (CO) toxicity may cause persistent injuries in tissues sensitive to hypoxia. Neuropsychiatric sequelae may be observed in about 67% of cases after severe CO exposure. AIM: The aims of this study were to demonstrate the usefulness of S-100beta and neuron-specific enolase (NSE) in CO intoxications, show the degree of neurological response, and determine the indications for hyperbaric oxygen treatment (HBOT) as biochemical markers. RESULTS: The S-100beta and NSE levels of the sera of 30 patients were studied upon admittance and at the third and sixth hours. S-100beta levels were found to be high in all 3 analyses. There was no significant change in NSE levels. When the S-100beta levels were compared with Glasgow Coma Scale levels, a strong negative correlation was found for all hours (r = -0.7, -0.8; P = .00). The correlation between S-100beta and carboxyhemoglobin levels at the initial hour was found to be statistically significant (r = 0.4; P = .01). The S-100beta levels in patients receiving HBOT showed a considerable decrease compared with those in patients not receiving the treatment. The same decrease was valid for NSE, although it was insignificant. CONCLUSION: S-100beta may be useful in evaluating intoxications as an early biochemical marker in CO intoxications, as well as in the differential diagnosis due to other causes, and in determining HBOT indications. PMID- 20006204 TI - Alcohol, bicycling, and head and brain injury: a study of impaired cyclists' riding patterns R1. AB - OBJECTIVE: The aim of the study was to examine the interactions between alcohol, bicycle helmet use, experience level, riding environment, head and brain injury, insurance status, and hospital charges in a medium-sized city without an adult helmet law. METHODS: A study of adult bicycle accident victims presenting to a regional trauma center over a 1-year period was undertaken. Data were collected at the bedside regarding helmet use, alcohol use, experience level, location and type of accident and prevailing vehicle speed (for road accidents), and presence and degree of head or brain injury. RESULTS: Two hundred patients 18 years or older were enrolled from December 2006 through November 2007. Alcohol use showed a strong correlation with head injury (odds ratio, 3.23; 95% confidence interval, 1.57-6.63; P = .001). Impaired riders were less experienced, less likely to have medical insurance, rarely wore helmets, were more likely to ride at night and in slower speed zones such as city streets, and their hospital charges were double (all P values <.05). CONCLUSIONS: Alcohol use leads to a host of unsafe bicycling practices, increased head and brain injuries, and costs to the cyclist and community. The interrelated characteristics of the riding patterns of the cyclists who use alcohol might help target interventions. PMID- 20006205 TI - Computed tomographic screening for thoracic and lumbar fractures: is spine reformatting necessary? AB - INTRODUCTION: Patients who sustain traumatic vertebral fractures often have multiple other associated injuries. Because of the mechanisms of injury, many of these patients routinely undergo chest computed tomographic (CCT) and/or abdominal/pelvic computed tomographic (APCT) scans to diagnose intrathoracic or intra-abdominal injuries. These scans are routinely reformatted to provide more detailed imaging of the spine. Although the patient does not incur more radiation, the charges associated with this are significant. This study compared the sensitivity of these CT modalities in detecting thoracolumbar spine fractures. METHODS: A retrospective chart review identified blunt trauma victims, admitted through the emergency department, with a discharge diagnosis of thoracic or lumbar spine fracture that received (1) a chest and T-spine CT, (2) an abdominal/pelvic and lumbar spine CT, or both. Final radiologic readings of these patients' CT scans were obtained, and the sensitivities of the different imaging methods were compared. Discharge diagnosis of spine fracture was considered the gold standard. RESULTS: One hundred seventy-six APCT scans with reformatting and 175 CCT scans with reformatting were available for comparison. There were 9 of 176 false-negative APCT scans vs 3/176 false-negative lumbar spine CT scans. There were 14/175 false-negative CCT scans vs 2/175 false-negative thoracic spine CT scans. The differences in sensitivity were significant (P < .001) for both comparisons. CONCLUSIONS: Reformatting of CCT and APCT scans gives improved sensitivity in the detection of thoracic and lumbar spine fractures in trauma patients. Future study looking at clinically significant fractures or those that change clinical management decisions may find that the reformatted images are not routinely needed as a screening tool. PMID- 20006207 TI - Ultrasound detection of guidewire position during central venous catheterization. AB - INTRODUCTION: Ultrasound guidance decreases complications of central venous catheter (CVC) placement, but risks of arterial puncture and inadvertent arterial catheter placement exist. Ultrasound-assisted detection of guidewire position in the internal jugular vein could predict correct catheter position before dilation and catheter placement. METHODS: Ultrasound examinations were performed in an attempt to identify the guidewire before dilation and catheter insertion in 20 adult patients requiring CVC placement. Central venous pressures were measured after completion of the procedure. RESULTS: Guidewires were visible within the lumen of the internal jugular vein in all subjects. Central venous pressures confirmed venous placement of catheters. Ultrasound visualization of the guidewire predicted venous CVC placement with 100% sensitivity (95% confidence interval 80-100%) and 100% specificity (95% confidence interval 80%-100%). CONCLUSIONS: Ultrasound reliably detects the guidewire during CVC placement and visualization of the wire before dilation and catheter insertion may provide an additional measure of safety during ultrasound-guided CVC placement. PMID- 20006206 TI - Ultrasound-guided femoral nerve blocks in elderly patients with hip fractures. AB - OBJECTIVES: The primary objective of this study was to determine the feasibility of ultrasound-guided femoral nerve blocks in elderly patients with hip fractures in the emergency department (ED). The secondary objective was to examine the effectiveness of this technique as an adjunct for pain control in the ED. METHODS: This prospective observational study enrolled a convenience sample of 13 patients with hip fractures. Ultrasound-guided femoral nerve block was performed on all participants. To determine feasibility, time to perform the procedure, number of attempts, and complications were measured. To determine effectiveness of pain control, numerical rating scores were assessed at baseline and at 15 minutes, 30 minutes, and hourly after the procedure for 4 hours. Summary statistics were calculated for feasibility measures. Wilcoxon matched-pairs signed-rank tests and Friedman analysis of variance test were used to compare differences in pain scores. RESULTS: The median age of the participants was 82 years (range, 67-94 years); 9 were female. The median time to perform the procedure was 8 minutes (range, 7-11 minutes). All procedures required only one attempt; there were no complications. After the procedure, there were 44% and 67% relative decreases in pain scores at 15 minutes (P < or = .002) and at 30 minutes (P < or = .001), respectively. Pain scores were unchanged from 30 minutes to 4 hours after the procedure (P < or = .77). CONCLUSIONS: Ultrasound-guided femoral nerve blocks are feasible to perform in the ED. Significant and sustained decreases in pain scores were achieved with this technique. PMID- 20006208 TI - Bajiaolian poisoning-a poisoning with high misdiagnostic rate. AB - BACKGROUND: One of the oldest Chinese herbal medicine, bajiaolian is widely used in traditional therapy. In Taiwan, bajiaolian is the fifth highest cause of poisoning among herbal medicines. The diagnosis is difficult because physicians are unfamiliar with this medicine's multiple presentations in different stages of intoxication. PROCEDURES: The records of 4 major poison centers in Taiwan were searched for all bajiaolian intoxication from July 1985 (the opening of first poison center) to March 2003. Two emergency physicians with toxicologic training reviewed the admission charts and visited case patients for follow-up. FINDINGS: Seventeen patients were identified, of which 15 (88.2%) had been misdiagnosed initially. In the beginning of their medical care, 14 cases were diagnosed as acute gastroenteritis. CONCLUSION: Bajiaolian intoxication is probably misdiagnosed because of early gastrointestinal symptoms followed by neurologic symptoms. A detailed patient history should be taken, and symptoms should be reviewed systemically to improve diagnostic accuracy. PMID- 20006209 TI - Use of midlevel providers in US EDs, 1993 to 2005: implications for the workforce. AB - OBJECTIVE: The aim of the study was to evaluate use of physician assistants (PAs) and nurse practitioners (NPs) in US emergency departments (EDs). METHODS: We analyzed visits from the 1993 to 2005 National Hospital Ambulatory Medical Care Survey, seen by midlevel provider (MLP), and compared characteristics of MLP visits to those seen by physicians only. RESULTS: From 1993 to 2005, 5.2% (95% CI, 4.6%-5.8%) of US ED visits were seen by PAs and 1.7% (95% CI, 1.5%-2.0%) by NPs. During the study period, PA visits rose from 2.9% to 9.1%, whereas NP visits rose from 1.1% to 3.8% (both P(trend) < .001). Compared to physician only visits, those seen only by MLPs arrived by ambulance less frequently (6.0% vs 15%), had lower urgent acuity (37% vs 59%), and were admitted less often (3.0% vs 13%). CONCLUSIONS: Midlevel provider use has increased in US EDs. Their involvement in some urgent visits and those requiring admission suggests that the role of MLPs extends beyond minor presentations. PMID- 20006210 TI - Termination of drug-induced torsades de pointes with overdrive pacing. AB - Drug-induced prolongation of the QT interval is frequently encountered after medication overdose. Such toxicity can result in degeneration to torsades de pointes (TdP) and require overdrive pacing. We present 3 cases in which intentional medication overdose resulted in QTc prolongation with subsequent degeneration to TdP. Despite appropriate care, including magnesium therapy, each case required overdrive pacing for resolution of TdP. Although rarely encountered, patients with drug-induced TdP can be successfully managed with overdrive pacing. PMID- 20006211 TI - Pyometra: a potentially lethal differential diagnosis in older women. PMID- 20006212 TI - McKittrick-Wheelock syndrome: is it really rare? PMID- 20006213 TI - If you see the contusion, there is no pneumothorax. PMID- 20006215 TI - Issues on percutaneous feeding tube replacement. PMID- 20006216 TI - Intraperitoneal versus retroperitoneal air signs in abdominal radiographs. PMID- 20006217 TI - First report of hypoglycemia secondary to dandelion (Taraxacum officinale) ingestion. PMID- 20006218 TI - Celiac artery dissection presenting with abdominal and chest pain. PMID- 20006219 TI - Stercoral perforation of colon: a rare but important mimicker of acute appendicitis. PMID- 20006220 TI - Negative pressure pulmonary hemorrhage induced by a candy. PMID- 20006221 TI - Successful treatment of traumatic coronary artery dissection with angiographic stenting. PMID- 20006222 TI - Severe reaction to inadvertent intravenous administration of a large dose of norepinephrine. PMID- 20006223 TI - Adult Still disease: worsening inflammatory changes in a 26-year-old woman. PMID- 20006224 TI - Ischemic stroke in trauma patients: investigating the source of embolus. PMID- 20006225 TI - Hypothermia with extracorporeal membrane oxygenation for sudden cardiac death and submersion. PMID- 20006226 TI - Primary torsion of omentum: a rare cause of acute abdomen. PMID- 20006227 TI - Major ST-segment elevation hiding acute severe pancreatitis. PMID- 20006228 TI - Management of foreign body in esophagus with rigid bronchoscopy. PMID- 20006229 TI - Intravenous thrombolysis in a patient with known cavernous malformation: a first case report. PMID- 20006230 TI - Rapid diagnosis of jejunojejunal intussusception by an emergency physician performed bedside ultrasound. PMID- 20006231 TI - Actinomycosis osteomylelitis of the mandible. PMID- 20006232 TI - Acute gastric volvulus: a rare but real surgical emergency. PMID- 20006233 TI - Medial wall fracture- induced pneumo-orbita mimicking inferior rectus entrapment. AB - The authors report 2 patients who experienced medial wall blowout fractures. Both patients presented with significant restriction of upgaze, mild proptosis, and crepitus of the upper lid. Computed tomography revealed significant pneumo-orbita filling the superior orbit with inferior displacement of the muscle cone and subcutaneous emphysema. No floor fractures were seen in either patient, but in both cases, the medial wall was breached and was almost certainly the source of the intraorbital air. Patients were managed conservatively, and the vertical gaze deficiencies resolved after 3 to 5 days. Large amounts of intraorbital and extraorbital air in the absence of a floor fracture can imitate inferior rectus entrapment and could potentially lead to unnecessary surgical intervention. PMID- 20006234 TI - Axillary artery transection after recurrent anterior shoulder dislocation. AB - Axillary artery transection after recurrent anterior shoulder dislocation is extremely rare. We present 2 such patients. The first, a 62-year-old man, presented with acute ischemia and a large hematoma in the axilla and chest wall. The second, a 63-year-old man, had a pseudoaneurysm and palpable peripheral pulses. Both underwent urgent computed tomography, which confirmed the clinical diagnosis, and the patients were taken to the operating room. In the first patient, intraoperative angiogram through both the brachial and the femoral route showed complete disruption of the axillary artery rendering an endovascular approach not possible. Proximal balloon occlusion was then undertaken through the femoral artery, controlling the bleeding and allowing easier dissection of the ruptured segment. Revascularization was performed with an interposition polytetrafluoroethylene (PTFE) bypass restoring normal blood supply to the upper extremity. The second patient had a Viabhan (W.L. Gore, Flagstaff, Ariz) stent graft implanted through the brachial artery with an excellent clinical and angiographic result. As expected, both patients had significant neurologic morbidity due to associated brachial plexus palsy. Ruptured axillary artery after shoulder dislocation is very uncommon. Endovascular repair and hybrid procedures combining open and endovascular techniques can offer reliable solutions to these challenging problems. PMID- 20006235 TI - Assessing response to changing plasma/red cell ratios in a bleeding trauma patient. AB - Recent military experience suggests that transfusing fresh frozen plasma and packed red cells in a 1:1 ratio may improve survival in exsanguinating trauma patients. We report the case of a single patient who required massive transfusion after suffering a single gunshot wound. Initially, the patient received FFP:PRBC in 1:2 ratio, but this did not correct laboratory parameters except for INR and clotting factor VII level, which were likely normalized by treatment with recombinant activated factor VII. After receiving FFP:PRBC in a 4:5 ratio, he continued to bleed and his coagulation profile showed no appreciable improvement. In the final phase, he received FFP:PRBC in a 7:5 ratio and his laboratory parameters of coagulopathy normalized, except for factor V level which was improved. He also clinically stopped bleeding. PMID- 20006236 TI - Soccer player whiplash maculopathy. PMID- 20006237 TI - Dimerized plasmin fragment D: a reliable biomarker for diagnosing aortic dissection? PMID- 20006238 TI - Leukocoria and irregular pupil. PMID- 20006239 TI - Wrist trauma. Preface. PMID- 20006240 TI - Wrist anatomy and surgical approaches. AB - Appreciation and knowledge of anatomy as it relates to surgical approaches is critical for planning treatment of traumatic wrist injuries. This article discusses the pertinent anatomy and some of the more commonly used approaches to wrist trauma. PMID- 20006241 TI - Physical examination of the wrist. AB - Physical examination of the wrist requires knowledge of wrist anatomy and pathology to make a diagnosis or narrow the differential diagnosis. Symptoms are provoked by palpation and signs are produced by manipulation. Negative findings elsewhere in the wrist are important. Final diagnosis may require diagnostic imaging. By having all three methods of assessment agree one is assured of correct diagnosis. The physical examination of the wrist is not unlike that of other joints, in that a systematic approach includes observation, range of motion, palpation, and special tests. PMID- 20006242 TI - Distal radius fractures--classification of treatment and indications for surgery. AB - Distal radius fractures are common injuries. Multiple classification systems have highlighted the evolution of the understanding of distal radius fractures. Understanding the classifications of distal radius fractures is important in identifying the important aspects that affect their outcome. Surgical indications of distal radius fractures can be divided into the following categories: patient factors, fracture reduction, fracture stability, and the presence of associated injuries. PMID- 20006243 TI - Distal radius fractures: nonoperative and percutaneous pinning treatment options. AB - Nonoperative treatment of distal radial fractures by reduction and immobilization remains the most common treatment, based on the incidence of appropriate fracture types, as seen in many epidemiological studies in the literature. In this article, the indications, technique, predictors of failure, outcomes, and complications are reviewed. A variety of treatment options have been proposed for distal radial fractures that are predicted, or subsequently identified, to be too unstable for nonoperative management. Percutaneous pinning is an effective option for select fractures. The authors also review the indications, described techniques, complications and outcomes associated with this treatment option. PMID- 20006244 TI - External fixation of distal radius fractures. AB - Fractures of the distal radius are the most common fractures that occur in patients between ages 15 and 75 years. Many methods for treating displaced distal radius fractures are available. All forms of treatment involve obtaining fracture reduction, which may then be maintained by casting, functional bracing, external fixation, percutaneous pinning, internal fixation, or a combination of these methods. This article discusses the indications and technique of fracture treatment with external fixation and, when required, adjuvant percutaneous pins. PMID- 20006245 TI - Plating for distal radius fractures. AB - No area of fracture management has had such a recent explosion of new treatment modalities as distal radius plating. This explosion has largely been implant- and industry-driven, with little evidence-based research guiding the way. A perceived difficulty with commonly used modalities by the orthopedic community has been enough to drive an entire new set of options for distal radius fixation. A drift from dorsal to volar plating has occurred that has been unexamined by randomized research. Segment specific fixation has been a new mindset that has resulted in a novel plate line and has caused other manufacturers to redesign their product lines. Other novel approaches for proposed problems include locking plates, nail plate combinations, and others. This article outlines some of these options with a literature opinion and a clarification from the authors. A treatment plan for common fractures of the distal radius is also outlined. PMID- 20006246 TI - Management of post-traumatic malunion of fractures of the distal radius. AB - Distal radius malunions are a common cause of patient morbidity. This review of the literature surrounding distal radius malunion covers the demographics, pathologic anatomy, and indications for surgery, surgical techniques, and salvage options. Particular emphasis is placed on subject areas that have not been reviewed as extensively in previous articles, including: intra-articular malunion, computer-assisted techniques, bone graft alternatives, and volar fixed angle plate osteosynthesis. PMID- 20006247 TI - Complications of distal radius fractures. AB - Fractures occur at the distal end of the radius more frequently than at any other location. The reported complication rates of distal radius fractures in the literature vary from 6% to 80%. Complications may occur from the fracture or its treatment. This article reviews complications caused by distal radius fractures and their treatment. Complications are divided chronologically into immediate, early (less than 6 weeks), and late (greater than 6 weeks). PMID- 20006248 TI - Acute scaphoid fractures. AB - Scaphoid fractures are a common problem encountered in clinical practice. This manuscript provides an algorithm for the diagnosis, evaluation, and treatment of acute scaphoid fractures. PMID- 20006249 TI - Management of scaphoid nonunions. AB - Scaphoid nonunions can exist with or without avascular necrosis of the proximal pole, and waist fractures may have an associated humpback deformity. CT best shows the deformity and bone loss, whereas MRI will show avascular necrosis. Operative treatment should be directed at correcting the deformity with open reduction and internal fixation and bone grafting. Vascularized bone grafts should be used in cases of avascular necrosis. PMID- 20006250 TI - Carpal bone fractures. AB - Carpal bone fractures make up a significant proportion of injuries to the wrist. The complex bone shape and articulations make diagnosis more difficult and missed injuries more common. This article reviews carpal bone fractures excluding the scaphoid. PMID- 20006251 TI - The diagnosis and treatment of scapholunate instability. AB - Scapholunate instability is the most common form of carpal instability. Pain produced by this condition is caused by the wrist's inability to sustain physiologic loads because of an injury to the linkage between the scaphoid and lunate. The term scapholunate instability may describe a wide spectrum of clinical conditions ranging from mild wrist dysfunction and partial ligamentous tear to debilitating pain with associated rupture of the scapholunate interosseus ligament complex. This article reviews the pathophysiology of scapholunate instability and its identification and treatment. PMID- 20006252 TI - Perilunate injuries. AB - Perilunate injuries are complex injuries of the bony and ligamentous structures of the wrist. They require operative management with careful restoration of carpal alignment and open reduction and internal fixation of associated fractures. Even with optimal treatment, mild to moderate dysfunction affects most patients. PMID- 20006253 TI - Traumatic injuries of the distal radioulnar joint. AB - Traumatic injuries of the distal radioulnar joint (DRUJ) may give rise to complex wrist pathologies. Substantial ongoing disability can arise should these injuries go unrecognized resulting in sub-optimal treatment and lack of appropriate rehabilitation. Injuries of the DRUJ may occur in isolation but more commonly are found with a fracture of the radius. These challenging DRUJ injuries may be simple or complex (irreducible or severe instability), acute or chronic. An adequate knowledge of the stabilizers of the DRUJ is essential in understanding treatment options. Traumatic instability of the DRUJ is reviewed and the anatomy and stabilizing factors are discussed. An algorithm to guide selection of treatment options in complex cases is presented. PMID- 20006254 TI - Low-dose spinal hyperbaric bupivacaine for adult anorectal surgery: a double blinded, randomized, controlled study. AB - BACKGROUND AND OBJECTIVE: To produce selective spinal anesthesia for adult anorectal surgery. STUDY DESIGN: Double-blinded, randomized, controlled trial. SETTING: Operating room and postoperative recovery area. PATIENTS: 152 adult, consecutive ASA physical status I, II, and III patients. INTERVENTIONS: After patients underwent dural puncture in the sitting position at L3-L4 or L4-L5, 0.5% hyperbaric bupivacaine was injected over two minutes: Group S7.5 received 1.5 mL, Group S5 received 1.0 mL, and Group S4 0.8 mL. After sitting for 10 minutes, patients were positioned for surgery. MEASUREMENTS: Rate of success, level and duration of sensory and motor block, time to voiding and ambulation, complications, and quality of anesthesia according to the patient and medical staff, were recorded. RESULTS: Spinal block had a 98% rate of success. Mean level of sensory block was 10.4 +/- 1.7, 7.4 +/- 2.2, and 7.0 +/- 1.8 dermatomes in Groups S7.5, S5, and S4 (P < 0.01 S7.5 vs S5, and S7.5 vs S4). Mean duration of sensory block was 310.5 +/- 42.6, 255.9 +/- 43.7, and 228.8 +/- 34.8 min in Groups S7.5, S5, and S4 (P < 0.01 S7.5 vs S5, S7.5 vs S4, and S5 vs S4). Motor block was Bromage score 2-3 in 70.5% of Group S7.5 patients versus Bromage score 0-1 in 97.3% of Group S5 and 92.1% of Group S4 patients (P < 0.05). CONCLUSION: A dose of 4 mg of hyperbaric bupivacaine produces a similar level of sensory and motor block as a 5 mg dose but with shorter duration and faster recovery. PMID- 20006255 TI - Spread of subarachnoid sensory block with hyperbaric bupivacaine in second trimester of pregnancy. AB - STUDY OBJECTIVE: To compare the spread of subarachnoid sensory block with hyperbaric bupivacaine in second trimester pregnant and non-pregnant women. DESIGN: Prospective study. SETTING: University teaching hospital. PATIENTS: 44 ASA physical status I and II women patients, 22 of whom were in their second trimester of pregnancy undergoing cervical cerclage, and 22 non-pregnant women scheduled for perianal surgery. INTERVENTIONS: The extent of sensory block and hemodynamic changes were assessed. MEASUREMENTS: Number of dermatomes blocked was determined by testing for pinprick; systolic blood pressure (SBP), diastolic blood pressure (DBP), and heart rate (HR) were measured at 3, 5, 10, 15, 30 and 60 minutes. MAIN RESULTS: Maximal sensory block was higher in the second trimester of the pregnant group by three dermatomes than the non-pregnant group. There were no statistically significant differences in SBP, DBP, or HR changes between the groups. CONCLUSION: Pregnant women in the second trimester exhibit enhanced spread of spinal analgesia with hyperbaric bupivacaine more so than non pregnant women. PMID- 20006256 TI - Intrathecal baclofen for postoperative analgesia after total knee arthroplasty. AB - STUDY OBJECTIVE: To determine whether intrathecal baclofen is an effective adjunctive agent to decrease acute and chronic postoperative pain after total knee arthroplasty. DESIGN: Prospective, randomized, double-blind controlled trial. SETTING: Operating room and inpatient units of a university hospital. PATIENTS: 60 adult, ASA physical status I, II, and III patients presenting for total knee arthroplasty. INTERVENTIONS: Anesthesia was provided by spinal injection of 15 mg of 0.75% hyperbaric bupivacaine combined with either 100 mcg baclofen or saline. Sedation was provided with intravenous midazolam and propofol. MEASUREMENTS: Data were collected on adverse effects, opioid usage, and verbal pain scale (VPS) from 0 to 10. The study period was divided into six discrete time intervals that included the 1(st) 72-hour postoperative period and a three-month post-discharge follow-up telephone call. MAIN RESULTS: The baclofen group used less morphine in the PACU than the control group (5 mg vs. 9.3 mg; P = 0.04). VPS were lower in the baclofen group than the treatment group, but significant differences could be demonstrated only in the time periods 48-72 hours and three months postoperatively. At three months, fewer patients in the baclofen group reported pain than the control group (8/27 vs. 19/29; P = 0.009). Regression analysis showed that the baclofen group was 4.5 times less likely to report pain at three months (95% CI: 1.5-16.6). CONCLUSIONS: IT baclofen used as an adjuvant to spinal anesthesia for total knee arthroplasty allows for less postoperative opioid usage and less chronic pain at three months. PMID- 20006257 TI - Comparison of patient-controlled analgesia with and without dexmedetomidine following spine surgery in children. AB - STUDY OBJECTIVE: To evaluate the effect of dexmedetomidine as an adjunct to patient-controlled analgesia (PCA) with morphine. DESIGN: Retrospective comparison. SETTING: University-affiliated children's hospital. MEASUREMENTS: The medical charts of 131 children with idiopathic scoliosis (IS) and NMS who had major spine surgery were reviewed. Out of 131, postoperatively 94 children received PCA with morphine alone (PCA group) and the remaining 37 children received PCA morphine and dexmedetomidine infusion at 0.4 mcg/kg/hour for 24 hours (PCA + Dex group). Preoperative, intraoperative, and postoperative morphine use data were collected. MAIN RESULTS: Intraoperative use of morphine was similar in children with IS and NMS. However, patients with IS used more morphine than patients with NMS on the first, second and third postoperative days in both groups. In children with IS, use of morphine on the second postoperative day was significantly higher in the PCA + Dex group (73 mg [50.5, 110.5]) than the PCA alone group (54 mg [36, 69], P = 0.03). The overall frequency of all perioperative complications was more in the PCA alone group (40% vs. 32%) than the PCA + Dex group. CONCLUSION: Postoperative 24-hour dexmedetomidine infusion as an adjunct to PCA with opioids might have a morphine-sparing effect as evidenced by the increase in morphine use on postoperative day 2 after the dexmedetomidine infusion was stopped. PMID- 20006258 TI - Aprotinin use during cardiac surgery: recent alterations and effects on blood product utilization. AB - OBJECTIVE: To investigate a single institution's changing use of aprotinin and subsequent effects on intraoperative blood product utilization (red blood cells/fresh frozen plasma) and postoperative clinical bleeding requiring reoperation. DESIGN: Retrospective study. SETTING: Single university institution (University of Chicago). MEASUREMENTS: Data from 499 adult patients undergoing cardiac surgery requiring cardiopulmonary bypass (CPB) over a two-year period (February 2005 - January 2007) were reviewed. The first 12 months (Feb 2005 - Jan 2006, Group 2005-2006) of data were compared with that from the second 12-month period (Feb 2006 - Jan 2007, Group 2006-2007). Information regarding patient demographics, surgical procedures, aprotinin use (none, half-dose, full-dose), and blood product use during CPB was retrospectively retrieved and analyzed. MAIN RESULTS: When Group 2006-2007 data was compared with that from Group 2005-2006, full-dose aprotinin use had significantly decreased (58% to 17%, P < 0.001), non use of aprotinin significantly increased (18% to 47%, P < 0.001), while fresh frozen plasma (FFP) utilization during CPB significantly increased (24% to 36%, P = 0.004). Red blood cell (RBC) transfusion rates remained stable (67% - 69%) yet rates of RBC and FFP transfusion during CPB significantly increased (23% to 34%, P = 0.003). There was also a trend toward increased unplanned reoperations for excessive clinical bleeding (0 pts in Group 2005-2006, three pts in Group 2006 2007). CONCLUSIONS: As the institution's use of high-dose aprotinin has significantly decreased, the number of patients requiring FFP and FFP/RBC combinations during CPB has significantly increased. Furthermore, a trend toward increasing incidence of unplanned reoperations for excessive clinical bleeding was noted. PMID- 20006259 TI - Should anesthesia residents with a history of substance abuse be allowed to continue training in clinical anesthesia? The results of a survey of anesthesia residency program directors. AB - STUDY OBJECTIVE: To determine the experience, attitudes, and opinions of program directors regarding the reintroduction of residents in recovery from substance abuse into the clinical practice of anesthesiology. DESIGN: Survey instrument. SETTING: Anesthesia residency training programs in the United States. MEASUREMENTS: After obtaining institutional review board approval, a list of current academic anesthesia residency programs in the United States was compiled. A survey was mailed to 131 program directors along with a self-addressed stamped return envelope to ensure anonymity. Returned surveys were reviewed and data compiled by hand, with categorical variables described as frequency and percentages. MAIN RESULTS: A total of 91 (69%) surveys were returned, representing experience with 11,293 residents over the ten-year period from July of 1997 through June of 2007. Fifty-six (62%) program directors reported experience with at least one resident requiring treatment for substance abuse. For residents allowed to continue with anesthesia residency training after treatment, the relapse rate was 29%. For those residents, death was the initial presentation of relapse in 10% of the reported cases. 43% of the program directors surveyed believe residents in recovery from addiction should be allowed to attempt re-entry while 30% believe that residents in recovery from addiction should not. CONCLUSIONS: The practice of allowing residents who have undergone treatment for substance abuse to return to their training program in clinical anesthesia remains highly controversial. They are often lost to follow-up, making it difficult, if not impossible to determine if re-training in a different medical specialty decreases their risk for relapse. A comprehensive assessment of the outcomes associated with alternatives to re-entry into clinical anesthesia training programs is needed. PMID- 20006260 TI - Intraoperative drug-eluting stent thrombosis in a patient undergoing robotic prostatectomy. AB - Insertion of drug-eluting stents is one of the strategies for treating patients with coronary artery disease. These patients can be a perioperative challenge in management as they need to be maintained on antiplatelet therapy to prevent stent thrombosis, which puts them at an increased risk for surgical bleeding. Recently revised guidelines on elective surgery following insertion of a drug-eluting stent recommend dual antiplatelet therapy for a period of twelve months. The management of a patient who presented for surgery more than two years after the insertion of a drug-eluting stent, and who developed in-stent thrombosis intraoperatively, is presented. PMID- 20006261 TI - Anesthetic management of a parturient with fetal sacrococcygeal teratoma and mirror syndrome complicated by elevated hCG and subsequent hyperthyroidism. AB - Mirror syndrome is a condition in which the mother "mirrors" her hydropic fetus and/or hydropic placenta. Physical and laboratory findings of mirror syndrome include generalized edema, hypertension, and proteinuria similar to preeclampsia. However, unlike preeclampsia, mirror syndrome is associated with hemodilutional anemia and fluid overload, which may progress to pulmonary edema. The anesthetic management of a parturient with fetal sacrococcygeal teratoma, hydrops fetalis, and mirror syndrome complicated by markedly elevated maternal serum human chorionic gonadotropin and subsequent clinical hyperthyroidism, is presented. PMID- 20006262 TI - Spinal epidural hematoma following epidural catheter removal in a paraplegic patient. AB - Spinal epidural hematoma is a rare and devastating complication of epidural catheter removal in an anticoagulated patient. The diagnosis could be quite challenging, especially in patients with preexisting neurological deficits. A 35 year-old patient with remote spinal cord injury and T4 level paraplegia developed a spinal epidural hematoma on the 7th postoperative day. The hematoma developed after epidural catheter removal with concurrent administration of unfractionated heparin. PMID- 20006263 TI - Anesthesia and airway management for removing pulmonary self-expanding metallic stents. AB - The use of bronchoscopically placed self-expanding metallic stents (SEMS) and silastic stents in patients suffering from tracheobronchial stenosis or similar problems has proven to be an important clinical option. When complications occur, it may be necessary to remove the device. Removal of a SEMS is usually performed during general anesthesia with muscle relaxation and positive pressure ventilation, often using total intravenous anesthesia. Airway management depends on stent type and location. Intubating patients' tracheas with a tracheal stent requires special caution, as it risks damaging tissue and dislodging the stent distally. Potential complications with removal include tracheal disruption, retained stent pieces, mucosal tears, re-obstruction requiring new stent placement, the need for postoperative ventilation, pneumothorax, damage to the pulmonary artery, and death. PMID- 20006264 TI - Ongoing paradoxical particulate embolism during megaprosthesis placement. AB - Pulmonary embolism (PE) by solid particles is an infrequent complication encountered during orthopedic procedures. A patent foramen ovale (PFO) allows embolic material into the left-sided circulation, potentially affecting several organs. A case of a 33-year-old woman undergoing total proximal femur resection with megaprosthesis reconstruction is presented. Towards the end of surgery, cement was injected to stabilize the prosthesis. Echodense particulate matter was first seen in the right atrium, subsequently in the right ventricle, and also in significant quantities in the left atrium and ventricle. This was accompanied by significant changes in partial pressure of arterial oxygen. Despite the aforementioned disturbances, hemodynamic stability was observed throughout the event. Postoperative follow-up showed persistent hypoxemia but no other abnormalities. This intriguing case of embolism via particulate material in a patient with PFO during hip replacement surgery allows review of the physiology and pathophysiology of PE phenomena. PMID- 20006265 TI - Cephalad migration of a thoracic epidural catheter in a 15-month old. PMID- 20006266 TI - Simulation saves resources. PMID- 20006267 TI - The use of ultrasound-guided hematoma blocks in wrist fractures. PMID- 20006268 TI - Closed-loop titration of propofol and remifentanil guided by Bispectral Index in a patient with extreme gigantism. PMID- 20006269 TI - Like methylene blue, indigo carmine might counteract vasoplegia: hemodynamic evidence obtained with pulse contour analysis. PMID- 20006270 TI - The societal cost of tuberculosis: Tarrant County, Texas, 2002. AB - PURPOSE: Cost analyses of tuberculosis (TB) in the United States have not included elements that may be prevented if TB were prevented, such as losses associated with TB-related disability, personal and other costs to society. Unmeasured TB costs lead to underestimates of the benefit of prevention and create conditions that could result in a resurgence of TB. We gathered data from Tarrant County, Texas, for 2002, to estimate the societal cost due to TB. METHODS: We estimated societal costs due to the presence or suspicion of TB using known variable and fixed costs incurred to all parties. These include costs for infrastructure; diagnostics and surveillance; inpatient and outpatient treatment of active, suspected, and latent TB infection (LTBI); epidemiologic activities; personal costs borne by patients and by others for lost time, disability, and death; and the cost of secondary transmission. A discount rate of 3% was used. RESULTS: During 2002, 108 TB cases were confirmed in Tarrant County, costing an estimated $40,574,953. The average societal cost per TB illness was $ 376,255. Secondary transmission created 47% and pulmonary impairment after TB created 35.4% of the total societal cost per illness. CONCLUSIONS: Prior estimates have concluded that treatment costs constitute most (86%) TB-related expenditures. From a societal perspective treatment and other direct costs account for little (3.3%) of the full burden. These data predict that preventing infection through earlier TB diagnosis and treatment of LTBI and expanding treatment of LTBI may be the most feasible strategies to reduce the cost of TB. PMID- 20006271 TI - Prepregnancy depressive symptoms and preterm birth in the Black Women's Health Study. AB - PURPOSE: To examine the association between prepregnancy depressive symptoms and preterm birth. METHODS: The present study is a prospective investigation of prepregnancy depressive symptoms-measured by the Center for Epidemiologic Studies Depression Scale (CES-D)-and risk of preterm birth reported in the Black Women's Health Study. With data on 2,627 singleton births (175 spontaneous and 163 medically indicated preterm births and 2,289 term births), we used generalized estimating equation models to estimate odds ratios (ORs) and 95% confidence intervals (CIs) adjusted for potential confounders. RESULTS: Relative to mothers with CES-D scores less than 16, the multivariable ORs of spontaneous preterm birth for mothers with CES-D scores of 16-22, 23-32, and > or =33 were 1.17 (95% CI = 0.78-1.80), 1.20 (95% CI = 0.69-2.10), and 2.00 (95% CI = 0.94-4.25), respectively (p for trend = 0.09). There was little evidence of an association between prepregnancy depressive symptoms and medically indicated preterm birth. CONCLUSIONS: Our data provide some evidence of an increased risk of spontaneous preterm birth among women with high prepregnancy depressive symptoms. PMID- 20006272 TI - Maternal pesticide exposure and neural tube defects in Mexican Americans. AB - PURPOSE: The relation between maternal pesticide exposures and neural tube defects (NTDs) in offspring was evaluated in 184 Mexican American case-women and 225 comparison women. METHODS: In-person interviews solicited information about environmental and occupational exposures to pesticides during the periconceptional period. RESULTS: With adjustment for maternal education, smoking, and folate intake, women who reported using pesticides in their homes or yards were two times more likely (95% confidence interval [CI], 1.2-3.1) to have NTD-affected pregnancies than women without these reported exposures. Case-women were also more likely to report living within 0.25 mile of cultivated fields than control-women (odds ratio [OR] 3.6; 95% CI, 1.7-7.6). As sources of pesticide exposure opportunities increased, risk of NTDs also increased. The adjusted ORs and 95% CIs for one, two, and three or more exposure sources were 1.2 (0.69-1.9), 2.3 (1.3-4.1) and 2.8 (1.2-6.3) respectively, and this positive trend was stronger for risk of anencephaly than for spina bifida. CONCLUSIONS: Self reported pesticide exposures were associated with NTD risk in this study population, especially use of pesticides within the home and a periconceptional residence within 0.25 mile of cultivated fields. PMID- 20006273 TI - U.S. Maternally linked birth records may be biased for Hispanics and other population groups. AB - PURPOSE: We sought to advance understanding of linkage error in U.S. maternally linked datasets and how the error might affect results of studies based on the linked data. METHODS: North Carolina birth and fetal death records for 1988-1997 were maternally linked (n=1,030,029). The maternal set probability, defined as the probability that all records assigned to the same maternal set do in fact represent events to the same woman, was used to assess differential maternal linkage error across race/ethnic groups. RESULTS: Maternal set probabilities were lower for records specifying Asian or Hispanic race/ethnicity, suggesting greater maternal linkage error for these sets. The lower probabilities for Hispanics were concentrated in women of Mexican origin who were not born in the United States. CONCLUSIONS: Differential linkage error may be a source of bias in studies that use U.S. maternally linked datasets to make comparisons between Hispanics and other groups or among Hispanic subgroups. Methods to quantify and adjust for this potential bias are needed. PMID- 20006274 TI - Urban-rural residence and the occurrence of cleft lip and cleft palate in Texas, 1999-2003. AB - PURPOSE: The etiology of orofacial clefts is complex and relatively unknown. Variation in cleft lip with or without palate (CLP) and cleft palate alone (CP) was examined in Texas across urban-rural residence (1999 to 2003). METHODS: Cases came from the Texas Birth Defects Registry (1,949 CLP and 1,054 CP) and denominator data came from vital records (254 counties; 1,827,317 live births). Variation in maternal residence was measured using four classification schemes: Rural Urban Continuum Codes, Urban Influence Codes, percentage of county in cropland, and Rural Urban Commuting Areas. Poisson regression was used to calculate rate ratios, adjusted for infant sex, plurality, gestational age, maternal parity, age, race/ethnicity, and education. RESULTS: Compared to the most urban referent category, living in more rural areas was associated with an increased adjusted risk of CLP. For example, the Rural-Urban Continuum Codes demonstrated elevated risks for CLP in "thinly populated areas" compared to "metropolitan-urban areas" (adjusted prevalence ratio = 1.9; 95% confidence intervals (CI) 1.2-2.8); CP was not similarly associated. Percentage of county cropland was not consistently associated with any outcome. CONCLUSION: The association patterns between non-urban residence and risk of CLP, except for percentage of cropland, suggests a constellation of exposures that may differ across urban-rural residence. PMID- 20006275 TI - Rising social inequalities in US childhood obesity, 2003-2007. AB - PURPOSE: This study examines changes between 2003 and 2007 in obesity and overweight prevalence among U.S. children and adolescents 10 to 17 years of age from detailed racial/ethnic and socioeconomic groups. METHODS: The 2003 (N=46,707) and 2007 (N=44,101) National Survey of Children's Health were used to calculate overweight and obesity prevalence (body mass index [BMI] > or = 85th and > or = 95th percentiles, respectively). Logistic regression was used to model odds of obesity. RESULTS: In 2007, 16.4% of U.S. children were obese and 31.6% were overweight. From 2003 to 2007, obesity prevalence increased by 10% for all U.S. children but increased by 23%-33% for children in low-education, low-income, and higher unemployment households. Obesity prevalence increased markedly among Hispanic children and children from single-mother households. In 2007, Hispanic, non-Hispanic Black, [corrected] and American Indian children had 3.0-3.8 times higher odds of obesity and overweight than Asian children; children from low income and low-education households had 3.4-4.3 times higher odds of obesity than children from higher socioeconomic households. The magnitude of racial/ethnic and socioeconomic disparities in obesity and overweight prevalence increased between 2003 and 2007, with substantial social inequalities persisting even after controlling for behavioral factors. CONCLUSIONS: Social inequalities in obesity and overweight prevalence increased because of more rapid increases in prevalence among children in lower socioeconomic groups. PMID- 20006276 TI - Distribution and cardiovascular risk correlates of plasma soluble intercellular adhesion molecule-1 levels in asymptomatic young adults from a biracial community: the Bogalusa Heart Study. AB - PURPOSE: That circulating soluble form of intercellular adhesion molecule-1 (sICAM-1) is associated with an increased risk for coronary artery disease is well recognized. However, information is scant regarding the distribution and cardiovascular (CV) risk correlates of sICAM-1 in asymptomatic young adults. METHODS: Plasma sICAM-1 was measured in 1,184 black and white persons in the Bogalusa Heart Study cohort (70% white, 43% male), aged 24 to 44 years. CV risk was assessed in terms of CV risk factors, status of parental CV disease, and composite carotid intima-media thickness (IMT). RESULTS: sICAM-1 levels displayed race difference (whites > blacks, p<0.0001), but no sex difference. In multivariate analysis including age, race, sex, smoking status, waist circumference, mean arterial pressure, low- and high-density lipoprotein (LDL and HDL) cholesterols, triglycerides, insulin resistance index, C-reactive protein (CRP), and adiponectin, the significant predictors of sICAM-1, in order of entry, were race (white > black), smoking, CRP, and waist circumference. Furthermore, there was a smoking by waist circumference interaction in that smoking attenuated the magnitude of correlation between waist circumference and sICAM-1. Levels of sICAM-1 adjusted for age, race, sex, and smoking increased with number of metabolic syndrome components (p for trend<0.01); positive family history of CV disease (p<0.05); and increased in composite carotid IMT specific for age, race, and sex (p for trend<0.05). CONCLUSION: These findings underscore the potential value of plasma sICAM-1 as an additional biomarker for CV risk among asymptomatic young adults. PMID- 20006277 TI - Assessing the quality of reporting of observational studies in cancer. AB - PURPOSE: The vast majority of epidemiological studies in cancer are observational. However, inadequate reporting of the published observational studies (OS) may restrict the generalizability and credibility of study results. The aim of this study was to evaluate the reporting quality of OS concerning cancer. METHODS: PubMed was searched for systematic reviews (SRs) of OS involving cancer published from January 2008 through February 2009. The citations provided in the SRs were evaluated for their reporting quality according to the STROBE (Strengthening the Reporting of Observational Studies in Epidemiology) statement, a checklist of items that are considered essential for good reporting of OS. The evaluation was focused on 25 methodological items/sub-items. The effect of journals' ranking in quality of reporting was also evaluated. RESULTS: The search identified 244 eligible OS included in seven SRs. Nine items/sub-items were reported by more than 90% of studies and 16 items/sub-items were reported by more than 70%. Some essential methodological aspects of OS (such as matching, absolute risks, missing data and flow diagram) were underreported. Significant differences were found among the seven SRs in the majority of the items. High and lower ranked journals were different only in reporting of results estimates and precision. CONCLUSIONS: The quality of reporting in OS in cancer was considered satisfactory, although certain items were underreported. Further improvement of reporting may enhance the validity of observational research. PMID- 20006278 TI - Is green tea drinking associated with a later onset of breast cancer? AB - BACKGROUND: Studies have found that tea polyphenols inhibit aromatase. Because of the substantial difference in levels of estrogens between premenopausal and postmenopausal women, the relationship between tea consumption and breast cancer risk may depend on menopausal status. METHODS: We examined this hypothesis in the Shanghai Women's Health Study, a population-based cohort study of 74,942 Chinese women. RESULTS: We found a time-dependent interaction between green tea consumption and age of breast cancer onset (p for interaction, 0.03). In comparison with non-tea drinkers, women who started tea-drinking at 25 years of age or younger had a hazard ratio (HR) of 0.69 (95% confidence interval [CI]: 0.41-1.17) to develop premenopausal breast cancer. On the other hand, compared with non-tea drinkers, women who started tea drinking at 25 years of age or younger had an increased risk of postmenopausal breast cancer with an HR of 1.61 (95% CI: 1.18-2.20). Additional analyses suggest regularly drinking green tea may delay the onset of breast cancer. CONCLUSIONS: Further studies are needed to confirm our findings. PMID- 20006279 TI - Serum calcium and breast cancer risk in a prospective cohort study. AB - Calcium has anti-proliferative and pro-differentiation effects on mammary cells in vitro and can inhibit the development of mammary tumors in mice. While there is some epidemiologic evidence for an inverse relation between dietary calcium intake and breast cancer risk, only one previous study has examined serum calcium levels in relation to breast cancer risk. We investigated this relation in a prospective cohort study of 2,762 women, aged 43-86, who were enrolled in the Beaver Dam Eye Study in 1988 and followed for up to 19 years. We found no evidence for an association between breast cancer risk and either total (hazard ratio, HR = 0.98; 95% CI: 0.60-1.60; 4(th) vs. 1(st) quartile) or ionized (HR = 0.85; 95% CI: 0.53, 1.38; 4(th) vs. 1(st) quartile) serum calcium levels. Additionally, we found no evidence for an association among subgroups defined by menopausal status and body mass index. PMID- 20006280 TI - Re: Number of 24-hour diet recalls needed to estimate energy intake. PMID- 20006282 TI - Has osteopathy a role to play in treatment of flu? PMID- 20006284 TI - Skin distraction at select landmarks on the spine midline in the upright and fully flexed postures. AB - BACKGROUND: This study was aimed at quantifying superoinferior and mediolateral skin distraction over the spine's midline for the purpose of designing a unique surface marker for use in a study originally proposed by Wojtys and Ashton Miller. It was also aimed at testing the null hypotheses H(01): There is no difference in the amount of skin distraction between hamstring normal and hamstring tight subjects and H(02): There are no age or gender differences of skin distraction. METHODS: Nine male and twelve female volunteers served as the convenience subjects for this IRB-approved study. Eight subjects were classified as hamstring "tight" (short) using the Finger-to-Floor Reach Test. Skin distraction was measured at five spine midline landmarks palpated on the subjects' bared backs: T1, T10, L3, S1, and the posterior-superior iliac spine (PSIS). A pattern of four dots was placed at each landmark using a rectangular template and non-toxic, water-soluble ink. Measurements were taken between superoinferior and mediolateral pairs of template points with subjects in both upright ("initial") and fully flexed ("final") postures. Between-measurement differences were then calculated, expressed as percent strain, and pooled for mean percent strain values. Repeated measures produced a maximum strain error of about 1.7%. RESULTS: With the exception of the skin over the T10 landmark, distraction in the superoinferior direction was greater than that in the mediolateral direction. There were no significant differences in skin distraction between age or gender groups. However, hamstring short males had significantly smaller superoinferior skin distraction at L3 than their hamstring normal counterparts [35% (+/-5.2) vs. 46% (+/-4.6), p=0.049), while hamstring short females had a smaller mean mediolateral distraction at the same level that approached significance [2.5% (+/-2.5) vs. 7.6% (+/-5.4), p=0.080). At this landmark, there was a moderately strong, inverse correlation (r=-0.720) between mediolateral percent strain and reach distance in hamstring tight subjects. CONCLUSION: In general, superoinferior percent strain increased and mediolateral percent strain decreased from thoracic to sacral regions, likely reflecting the relative increase in spine segment motion from thoracic to lumbar region. The larger mean mediolateral distraction at the T10 level was probably the result of traction on the skin at that level by the dependence of appendicular structures in forward flexion. Finally, the negative value at the T1 landmark was probably the result of the cervical spine extension that occurred during flexion as the subjects lifted their heads to look up. PMID- 20006283 TI - Fascia: A missing link in our understanding of the pathology of fibromyalgia. AB - Significant evidence exists for central sensitization in fibromyalgia, however the cause of this process in fibromyalgia-and how it relates to other known abnormalities in fibromyalgia-remains unclear. Central sensitization occurs when persistent nociceptive input leads to increased excitability in the dorsal horn neurons of the spinal cord. In this hyperexcited state, spinal cord neurons produce an enhanced responsiveness to noxious stimulation, and even to formerly innocuous stimulation. No definite evidence of muscle pathology in fibromyalgia has been found. However, there is some evidence for dysfunction of the intramuscular connective tissue, or fascia, in fibromyalgia. This paper proposes that inflammation of the fascia is the source of peripheral nociceptive input that leads to central sensitization in fibromyalgia. The fascial dysfunction is proposed to be due to inadequate growth hormone production and HPA axis dysfunction in fibromyalgia. Fascia is richly innervated, and the major cell of the fascia, the fibroblast, has been shown to secrete pro-inflammatory cytokines, particularly IL-6, in response to strain. Recent biopsy studies using immuno histochemical staining techniques have found increased levels of collagen and inflammatory mediators in the connective tissue surrounding the muscle cells in fibromyalgia patients. The inflammation of the fascia is similar to that described in conditions such as plantar fasciitis and lateral epicondylitis, and may be better described as a dysfunctional healing response. This may explain why NSAIDs and oral steroids have not been found effective in fibromyalgia. Inflammation and dysfunction of the fascia may lead to central sensitization in fibromyalgia. If this hypothesis is confirmed, it could significantly expand treatment options to include manual therapies directed at the fascia such as Rolfing and myofascial release, and direct further research on the peripheral pathology in fibromyalgia to the fascia. PMID- 20006285 TI - Relationship between hand-grip isometric strength and isokinetic moment data of the shoulder stabilisers. AB - The purpose of this study was to examine the relationship between hand-grip (HG) isometric strength and isokinetic moment of the shoulder musculature in 18 healthy male volunteers. HG isometric strength at 0 degrees , 90 degrees and 180 degrees of shoulder flexion and isokinetic peak and average concentric moments of the shoulder rotators and abductors and the elbow flexors at 60 degrees s(-1) were measured on both the dominant and non-dominant sides. Pearson correlation coefficients revealed statistically significant positive relationships between HG isometric strength and isokinetic moments of the shoulder external rotators (r=0.40-0.54), the shoulder abductors (r=0.42-0.71) and the elbow flexors (r=0.45 0.66) regardless of hand dominance. The positive relationships between HG isometric strength and isokinetic strength of the shoulder stabilisers was probably attributed to mechanisms providing stability to the elbow and shoulder joints either by force transmission via myotendinous and myofascial pathways or by "overflow" of muscular activity via neural circuits. The results of the present findings suggested that HG isometric strength can be used to monitor isokinetic strength of certain muscle groups contributing to the stability of the shoulder joint; however, HG strength may account only for approximately 16-50% of the variability in isokinetic strength of these muscle groups. PMID- 20006286 TI - Changes in the structure of collagen distribution in the skin caused by a manual technique. AB - OBJECTIVE: When treating patients with functional disorders using a special manual technique, tissue changes can be felt by the therapist and the patient. This study was conducted to objectively document these changes. METHOD: In the author's practice for body therapy, 30 patients were measured with high-frequency ultrasound (22MHz) immediately before and after their first treatment in the area where they experienced pain or other discomfort and/or movement restriction. RESULTS: Highly significant differences can be seen in the structure of the collagen matrix in the dermis before and after treatment. These changes reflect the differences in tension, softness and regularity, which can be palpated before and after treatment and are thought to be caused by changes in the mechanical forces of fibroblasts and increased microcirculation. PMID- 20006287 TI - Glove use and the HIV positive massage therapy client. AB - Massage therapy is often used to treat stress, and other symptoms of HIV/AIDS. Massage therapy standards of practice require the use of gloves only when contact with blood and bodily fluids is expected. Health care professionals often mistrust universal precautions and use gloves when their use is not indicated, especially when dealing with HIV positive clients. This case report explored the effects of un-indicated glove use on stress levels, satisfaction with treatment, perception of the therapist, and perceived stigma during a massage therapy treatment. In this case, gloved treatments were only 80% as effective at reducing stress as ungloved treatments. No difference was found in sense of stigma, perception of the therapist, or overall satisfaction in ungloved compared to gloved treatments. Suggestions for future considerations and additional research are made. PMID- 20006288 TI - Range of normality versus range of motion: A functional measure for the prevention and management of low back injury. AB - The Range of Motion (ROM) is a popular measurement used in the determination of disability for low back pain subjects in spite of serious objections to its clinical usefulness. It is proposed to consider a different index called the Range of Normality (RON) which is defined to be the portion of the ROM that an injured subject is able to do quasi-normally. This permits a direct assessment of the return to work parameters and the restrictions that ought to be placed on activities. It also allows follow up since the RON is expected to fill up the ROM as the injured subject recovers from his injury. PMID- 20006289 TI - Functional fitness improvements after a worksite-based yoga initiative. AB - This study explored the benefits of yoga on functional fitness, flexibility, and perceived stress. A quasi-experimental design was used to measure benefits of yoga in sample of firefighters from a major metropolitan fire department. Yoga classes were conducted on-shift, in the fire stations over the period of 6 weeks. The classes included pranayama (breathing), asana (postures), and savasana (relaxation); 108 firefighters enrolled in the study, most were physically active but had no prior experience with yoga. Baseline and post-yoga assessments were completed by 77 participants. Paired t-tests revealed significant improvements in the Functional Movement Screen, a seven item test that measures functional fitness. Improvements were also noted in trunk flexibility and perceived stress. Participants also reported favorable perceptions of yoga: feeling more focused and less musculoskeletal pain. These findings - along with the retention of the majority of the participants - indicate that participants benefited from yoga. PMID- 20006290 TI - Effects of Anma therapy (traditional Japanese massage) on body and mind. AB - INTRODUCTION: Anma therapy is a traditional style of Japanese massage, one of touch and manual therapies, and one of the most popular CAM therapies in Japan. It was brought from China in the 6th century and, while based on the theory of Chinese medicine, it developed in Japan according to Japanese preference and has recently come to include theories of Western medicine. The purpose of this study was to clarify the physical and psychological effects of Anma therapy. PARTICIPANTS AND METHODS: Fifteen healthy female volunteers in their fifth decade, with chronic muscle stiffness in the neck and shoulder, received two interventions: 40-min Anma therapy and 40-min rest intervention. The design was cross-over design. Participants were randomly divided into two groups. Group A was started on Anma therapy from the first day followed by the rest intervention after a 3-day interval. The order of the Anma therapy and the rest intervention reversed for Group B. Visual Analogue Scale (VAS) score for muscle stiffness in the neck and shoulder, state anxiety score, and salivary cortisol concentration levels and secretory immunoglobulin A (s-IgA) were measured pre- and post interventions. RESULTS: Anma therapy significantly reduced VAS scores and state anxiety scores. S-IgA concentration levels increased significantly across both groups. CONCLUSION: Anma therapy reduced muscle stiffness in the neck and shoulder and anxiety levels in this pilot study of 50-year-old females. PMID- 20006291 TI - Performing arts medicine--instrumentalist musicians, Part II--examination. AB - Part I of this article's series included background information on performing arts medicine with a special focus on musicians. It covered in detail what questions need to be included in the history, when healthcare providers first examine musicians. In part II of the series, the emphasis is on the physical examination, including posture, range of motion and hypermobility, ergonomics, and instrument-specific examination procedures. The final article in the series will describe three case histories of musicians with hand pain. PMID- 20006292 TI - Qi Gong's relationship to educational kinesiology: A qualitative approach. AB - This paper qualitatively reviews two complementary therapies; Qi Gong and educational kinesiology (EK). It is being suggested that Qi Gong and EK may be united through a qualitative convergence and a shared underlying concept. The authors hypothesize that a coherent rationale can be formed through this conceptual synthesis and propose that to some extent Qi Gong movements and EK can be considered to work in unison with each other. The logical synthesis of these two therapies is being presented to identify Qi Gong movements with concepts of brain gymnastics and also to explain how this new construct can be developed and implemented into practice. When verified, this hypothesis will allow individuals to better understand Chinese health exercises from the modern science perspective such as neuroanatomy, neurophysiology and psychoneuroimmunology. PMID- 20006293 TI - Core stability is a subset of motor control. PMID- 20006294 TI - The myth of core stability. AB - The principle of core stability has gained wide acceptance in training for the prevention of injury and as a treatment modality for rehabilitation of various musculoskeletal conditions in particular of the lower back. There has been surprisingly little criticism of this approach up to date. This article re examines the original findings and the principles of core stability/spinal stabilisation approaches and how well they fare within the wider knowledge of motor control, prevention of injury and rehabilitation of neuromuscular and musculoskeletal systems following injury. PMID- 20006295 TI - The missing link in protecting against back pain. PMID- 20006296 TI - A summary of the Texas Tuberculosis Research Symposium 2009: emerging directions in TB research and clinical affairs. PMID- 20006297 TI - Pulmonary surfactant and tuberculosis. AB - Mycobacterium tuberculosis comes in contact with pulmonary surfactant, alveolar macrophages and type II epithelial cells. Alveolar type II epithelial cells secrete pulmonary surfactant, a complex mixture of phospholipids and proteins lining the alveolar surface, while alveolar macrophages are involved in surfactant catabolism. Surfactant proteins SP-A and SP-D modulate phagocytosis of M. tuberculosis by alveolar macrophages. We have reported that mice with decreased surfactant catabolism resulting from GM-CSF deficiency are highly susceptible to acute aerosol infection with 100 cfu of M. tuberculosis. Here, we evaluated the lungs of WT, GM-CSF-deficient, and GM-CSF-corrected mice surviving six months after sub-acute aerosol infection of 5-10 cfu M. tuberculosis. We show that GM-CSF-deficient mice develop intra-bronchial and intra-alveolar tuberculosis lesions with numerous mycobacteria, inflammatory cells, and extracellular proteinaceous material containing surfactant protein B (SP-B). In contrast, WT and GM-CSF-corrected mice develop typical epithelioid granulomas containing lymphocytes, SP-B positive cells, and M. tuberculosis bacilli inside macrophages. Our findings support the concept that whole pulmonary surfactant is an important component of host mycobacterial infection in the distal lung. PMID- 20006298 TI - Application of optical imaging to study of extrapulmonary spread by tuberculosis. AB - The incidence of extrapulmonary tuberculosis is increasing, possibly due to the high frequency of co-infection with HIV. Extrapulmonary infections complicate diagnosis, have higher mortality rates and are more difficult to treat. Insight into the mechanisms involved in extrapulmonary spread of tuberculosis is critical to improving management. We set out to better understand extrapulmonary spread kinetics in mice and guinea pigs as well as the effects of infectious dose. We found that extrapulmonary spread occurs at a discrete time point when infected by low-dose aerosol, but at high-dose aerosol it occurs within the first 24h. The ability to follow tuberculosis in real-time during infection would allow us to better address the mechanisms involved. We found that mycobacteria can be optically imaged after pulmonary infection in the mouse lung, suggesting that this technology could be applied to study of extrapulmonary spread of tuberculosis. PMID- 20006299 TI - TB research at UT-Houston--a review of cord factor: new approaches to drugs, vaccines and the pathogenesis of tuberculosis. AB - Tuberculosis remains a major threat as drug resistance continues to increase. Pulmonary tuberculosis in adults is responsible for 80% of clinical cases and nearly 100% of transmission of infection. Unfortunately, since we have no animal models of adult type pulmonary tuberculosis, the most important type of disease remains largely out of reach of modern science and many fundamental questions remain unanswered. This paper reviews research dating back to the 1950's providing compelling evidence that cord factor (trehalose 6,6 dimycolate [TDM]) is essential for understanding tuberculosis. However, the original papers by Bloch and Noll were too far ahead of their time to have immediate impact. We can now recognize that the physical and biologic properties of cord factor are unprecedented in science, especially its ability to switch between two sets of biologic activities with changes in conformation. While TDM remains on organisms, it protects them from killing within macrophages, reduces antibiotic effectiveness and inhibits the stimulation of protective immune responses. If it comes off organisms and associates with lipid, TDM becomes a driver of tissue damage and necrosis. Studies emanating from cord factor research have produced (1) a rationale for improving vaccines, (2) an approach to new drugs that overcome natural resistance to antibiotics, (3) models of caseating granulomas that reproduce multiple manifestations of human tuberculosis. (4) evidence that TDM is a key T cell antigen in destructive lesions of tuberculosis, and (5) a new understanding of the pathology and pathogenesis of postprimary tuberculosis that can guide more informative studies of long standing mysteries of tuberculosis. PMID- 20006300 TI - Methionine sulfoxide reductase B (MsrB) of Mycobacterium smegmatis plays a limited role in resisting oxidative stress. AB - Pathogenic mycobacteria including Mycobacterium tuberculosis resists phagocyte generated reactive oxygen intermediates (ROI) and this constitutes an important virulence mechanism. We have previously reported, using Mycobacterium smegmatis as a model to identify the bacterial components that resist intracellular ROI, that an antioxidant methionine sulfoxide reductase A (MsrA) plays a critical role in this process. In this study, we report the role of methionine sulfoxide reductase B (MsrB) in resistance to ROI by constructing a msrB mutant (MSDeltamsrB) and MsrA/B double mutant (MSDeltamsrA/B) strains of M. smegmatis and testing their survival in unactivated and interferon gamma activated mouse macrophages. WhilemsrB mutant exhibited significantly lower intracellular survival than its wild type counterpart, the survival rate seemed to be much higher than msrA mutant (MSDeltamsrA) strain. Further, the msrB mutant showed no sensitivity to oxidants in vitro. The msrA/B double mutant (MSDeltamsrA/B), on the other hand, exhibited a phenotype similar to that of msrA mutant in terms of both intracellular survival and sensitivity to oxidants. We conclude, therefore, that MsrB of M. smegmatis plays only a limited role in resisting intracellular and in vitro ROI. PMID- 20006301 TI - Tuberculosis in Texas 2008. PMID- 20006302 TI - Efficacy of Artemisia afra phytotherapy in experimental tuberculosis. AB - Artemisia afra [Jacq] (Asteraceae) phytotherapy is widely used for its medicinal properties in traditional practices. In this study we investigated whether extracts of A. afra are capable of controlling mycobacterial replication. For Mycobacterium aurum cultured in the presence of aqueous-, methanol- and dichloromethane (DCM) extracts of A. afra we found that bacterial replication was inhibited by the dichloromethane extract only. Activity of the DCM extract was confirmed in dose-dependent studies against both M. aurum and M. tuberculosis with an IC(50) =270 microg/ml and IC(50) = 290microg/ml, respectively. Fractionation of the DCM extract and evaluation of its efficacy in vitro found that most of the antimycobacterial activity was associated with isolate fraction C8 that contained several sesquiterpene lactones, the most prominent of which are Artemin and Arsubin. Evaluation of the bactericidal efficacy in vitro showed that isolate fraction C8 reduced replication of M. aurum and M. tuberculosis in a dose dependent manner with IC(50) =1.9 microg/ml and IC(50) = 2.0 microg/ml, respectively, and an MIC = 10 microg/ml. Further, isolate fraction C8 and the DCM extract was administered to M. tuberculosis-infected mice at a tolerated dose of 1000 microg/kg for up to 26 weeks and mycobacterial burdens compared to untreated , INH/RIF treated- and aqueous-extract-treated animals to assess its bactericidal activity in vivo. Bacterial replication remained unaffected during treatment with either isolate fraction C8 or the DCM extract resulting in pulmonary and splenic bacilli burdens comparable to that of untreated mice. In contrast, INH/RIF treatment cleared M. tuberculosis infection after only 8 weeks to undetectable levels. Interestingly, treatment of M. tuberculosis-infected mice with aqueous extract of A. afra regulated pulmonary inflammation during early infection notwithstanding its inability to inhibit mycobacterial growth. This study clearly demonstrates that A. afra contains in vitro anti-mycobacterial activity, modulates pulmonary inflammation in early mycobacterial infection, and that the mouse experimental tuberculosis model may serve as a useful assay for evaluating the utility of phytotherapy. PMID- 20006303 TI - Latent tuberculosis infections in hard-to-reach drug using population-detection, prevention and control. AB - Interferon-gamma release assays (IGRAs) need be evaluated for effectiveness as screening tests for tuberculosis (TB) infection in drug users. These tests have demonstrated improved sensitivity and specificity, but have not been studied in drug users. These one step blood tests are intended to replace the tuberculin skin test (TST), which is difficult to use and requires 48 hour follow-up, so they are expected to be particularly suitable for risk groups, like drug users, in whom follow-up is problematic. Drug users have traditionally been identified as being at increased risk for acquiring TB disease. The results of our pilot study using the TST and simpler and more sensitive interferon-gamma release assays showed that about 45% of current drug users in Houston tested have at least one test positive for latent tuberculosis infection (LTBI). These preliminary data suggest that there is an important reservoir of LTBI in drug using populations, and the risk of progression to active TB disease with other infections is great. However, LTBI in drug using populations has not been studied in depth and deserves further investigation. We need to evaluate the validity of IGRAs for detection of latent TB infection, the factors associated with LTBI, the incidence and risk for developing active TB disease in drug users and the effectiveness of early treatment of LTBI. We believe that using better tuberculosis screening tools will allow us to more accurately measure the prevalence of latent TB infection and incidence of active TB disease in drug using populations and develop more effective TB prevention and treatment interventions in the community. PMID- 20006304 TI - Anti-tuberculosis immunity induced in mice by vaccination with Mycobacterium smegmatis over-expressing Antigen 85B is due to the increased influx of IFNgamma positive CD4 T cells into the lungs. AB - BCG vaccine is unsafe for use in patients with AIDS. Mycobacterium smegmatis (Msm), an avirulent species unlike virulent Mycobacterium tuberculosis (H37Rv, Mtb) has been used as a carrier vaccine with ambiguous results due to the elicitation of poor immune responses to antigens in mice. In this study, we over expressed the immunodominant antigen 85B in M. smegmatis (Msm-OEAg85B) and compared the immunogenicity of Msm-OEAg85B with that of wild-type Msm. Mice which were vaccinated with either Msm or Msm-OEAg85B and challenged 2 weeks later with Mtb. Vaccine-induced protection and lung T cell responses were evaluated post vaccination and post challenge. Unlike wild-type Msm that elicited minimal T cell responses in mice, MsmOE-Ag85B induced enhanced CD4+IFNgamma+ T cell responses that leveled off over 2 weeks. After virulent challenge at 2 weeks, Mtb grew progressively in the lungs of naive mice and mice vaccinated with wild-type Msm, but showed reduced growth (<0.6 log(10)) and therefore protection in Msm-OEAg85B vaccinated mice. Lungs of Msm-OEAg85B-vaccinated mice showed increased numbers of CD4+IFNgamma+ T cells suggesting that the reduced bacterial growth was likely due to the enhanced T cell response in lungs. Since wild-type Msm was unable to protect but Msm-OEAg85B was, we suggest that Msm can be genetically manipulated to over-express selected Mtb antigens, thereby paving the way for safer vaccines that can be used in immunodeficient patients. PMID- 20006305 TI - Lactoferrin enhances efficacy of the BCG vaccine: comparison between two inbred mice strains (C57BL/6 and BALB/c). AB - The current vaccine for tuberculosis (TB), an attenuated strain of Mycobacterium bovis Bacillus Calmette Guerin (BCG), is effective to prevent childhood onset of the disease, but its efficacy is reduced in adults. One strategy to improve the existing vaccine is to develop more effective adjuvants. Lactoferrin, an iron binding glycoprotein possessing immune modulatory activities, is a promising adjuvant candidate. The studies presented here examine the effect of lactoferrin to enhance efficacy of the BCG vaccine using a vaccination/challenge protocol (8 weeks boost and challenge at 12 weeks post-boost) that focuses on reduction in development of pathological changes to lung tissue. C57BL/6 and BALB/c mice vaccinated with BCG/lactoferrin exhibited protection upon Mycobacterium tuberculosis (MTB) challenge, showing reduced pulmonary disease pathology and decreased organ bacterial load. In addition, BCG/lactoferrin-treated macrophages isolated from BALB/c mice, which express a relative reduced T(H)1 phenotypic response to MTB antigens compared to the C57BL/6 mouse, were able to activate a higher percentage of IFN-gamma-producing CD4+ splenocytes. Overall, lactoferrin stands as an adjuvant capable of enhancing efficacy of the BCG vaccine through induction of T(H)1 immune responses, even in hosts typically demonstrative of reduced T(H)1 responsiveness to BCG antigens. PMID- 20006306 TI - Progress in understanding the human immune responses to Mycobacterium tuberculosis. AB - Development of an effective vaccine against tuberculosis hinges on an improved understanding of the human immune response to Mycobacterium tuberculosis. Work in this area at the University of Texas Health Science Center at Tyler has led to advances in four areas: (1) natural killer cells contribute to innate immunity by lysing M. tuberculosis-infected mononuclear phagocytes, and to adaptive immunity by enhancing the CD8+ T-cell effector function and inhibiting expansion of T regulatory cells; (2) Interferon-gamma plays a central role in resistance to many intracellular pathogens, including M. tuberculosis, and we have identified three transcription factors that bind to the Interferon-gamma proximal promoter and increase Interferon-gamma transcription in live T-cells that are activated by M. tuberculosis antigens; (3) A DNA vaccine that encodes the M. tuberculosis 10fts;kDa culture filtrate protein and the lysosomal integral membrane protein-2 was produced to direct vaccine antigens to the MHC class II processing and presentation pathway. When this vaccine was coated with polyethylenimine and administered to mice, it yielded a remarkably potent pulmonary immune response that reduced the bacillary burden by 90% after M. tuberculosis challenge; (4) The early secreted antigenic target of 6fts;kDa (ESAT-6) is a putative vaccine antigen. We found that high concentrations of this antigen markedly inhibit Interferon-gamma production by T-cells and are working to understand the molecular mechanisms underlying this effect. Developing methods to enhance NK cell functions that favor protective immunity, increase interferon-gamma transcription, elicit protective pulmonary immune responses and prevent ESAT-6 from inhibiting T-cell function will contribute significantly to development of antituberculosis vaccines. PMID- 20006307 TI - Targeting the role of N-terminal methionine processing enzymes in Mycobacterium tuberculosis. AB - The discovery of anti-tuberculosis agents that target new pathways is crucial for effective short-term TB therapy that will limit the development of resistance. The clinical significance of multidrug-resistant and extensively drug-resistant strains of Mycobacterium tuberculosis, latent TB and Human Immunodeficiency Virus co-infection in tuberculosis patients have made the development of new antimycobacterials more imperative. A better understanding of the major pathways that are involved in the pathogenesis, survival, and dormancy of Mtb will aid in the identification of new drug targets. Here, we review the N-terminal methionine excision (NME) pathway as a potential drug target during host infection with M. tuberculosis. The removal of the N-terminal methionine is a requirement for some proteins prior to post-translational modifications and processing. Therefore, an understanding of the physiological relevance of the two families of enzymes at the center of NME - peptide deformylases and methionine aminopeptidases - has the prospect of adding novel targets and antimycobacterials to the pipeline. PMID- 20006308 TI - Mycobacterium tuberculosis ftsZ expression and minimal promoter activity. AB - Optimal levels of ftsZ gene product are shown to be required for initiation of the cell division process in Mycobacterium tuberculosis. Here, we report that the ftsZ gene expression is sharply down-regulated during starvation and hypoxia, conditions that are believed to result in growth arrest, but is restored upon dilution of cultures into fresh oxygen-rich media. Primer extension analysis identified four transcriptional start sites, designated as P1, P2, P3 and P4 at nucleotide positions -43, -101, -263, and -787, respectively, in the immediate upstream flanking region of the ftsZ initiation codon. Promoter deletion and homologous recombination experiments revealed that ftsZ expression from the 101 bp region is sufficient for M. tuberculosis viability. All promoter strains had reduced FtsZ levels compared to wild-type, although the loss of P4 severely compromised FtsZ levels during both the active and stationary phases. We propose that ftsZ expression from all promoters is required for optimal intracellular FtsZ levels and that the activities of P4 and possibly other promoters are down regulated during growth-arrest conditions. PMID- 20006309 TI - Overproduction and localization of Mycobacterium tuberculosis ParA and ParB proteins. AB - The ParA and ParB family proteins are required for accurate partitioning of replicated chromosomes. The Mycobacterium tuberculosis genome contains parB, parA and two parA homologs, Rv1708 and Rv3213c. It is unknown if parA and its homologs are functionally related. To understand the roles of ParA and ParB proteins in M. tuberculosis cell cycle, we have evaluated the consequences of their overproduction and visualized their localization patterns in M. smegmatis. We show that cells overproducing ParA, Rv1708 and Rv3213c and ParB are filamentous and multinucleoidal indicating defects in cell-cycle progression. Visualization of green-fluorescent protein fusions of ParA and its homologues showed similar localization patterns with foci at poles, quarter-cell, midcell positions and spiral-like structures indicating that they are functionally related. On the other hand, the ParB- GFP fusion protein localized only to the cell poles. The cyan- and yellow-fluorescent fusion proteins of ParA and ParB, respectively, colocalized at the cell poles indicating that these proteins interact and possibly associate with the chromosomal origin of replication. Collectively our results suggest that the M. tuberculosis Par proteins play important roles in cell-cycle progression. PMID- 20006310 TI - Mycobacterium tuberculosis ftsH expression in response to stress and viability. AB - FtsH is an essential membrane-bound protease that degrades integral membrane proteins as well as cytoplasmic proteins. We show that Mycobacterium tuberculosis (Mtb) ftsH expression levels are upregulated upon exposure to agents that produce reactive oxygen and nitrogen intermediates (ROI and RNI) and growth in macrophages. In partial support of this result is our observation that the Mtb merodiploid overexpressing ftsH shows increased resistance to ROI. ftsH transcript levels are downregulated during stationary phase and starvation. ftsH overexpression strain shows delayed growth and reduced viability in vitro and ex vivo. Finally, we show that the intracellular levels of FtsZ, an essential cell division protein, are reduced in ftsH-overexpressing strain. Together, our results suggest that Mtb FtsH is a stress-response protein that promotes the pathogen's ability to deal with ROI stress and is possibly involved in the regulation of FtsZ levels. PMID- 20006311 TI - Mycobacterium tuberculosis ESX-1 system-secreted protein ESAT-6 but not CFP10 inhibits human T-cell immune responses. AB - The secreted proteins of M. tuberculosis, early secreted antigenic target 6 kDa (ESAT-6) and culture filtrate protein 10 kDa (CFP10), have been identified as antigenic proteins with potent T-cell stimulatory effects, and therefore have been the focus of tuberculosis vaccine studies. However, recent work showed that secretion of these proteins by the specialized ESAT-6 secretion system (ESX)-1 of M. tuberculosis is associated with virulence and pathogenesis. The studies demonstrated that ESAT-6 inhibits antigen-presenting cell function by reducing IL 12 production by macrophages through interrupting TLR2 signaling pathways and inducing macrophage apoptosis. However, the effect of ESAT-6 on T cells remains unexplored. To address this question, we studied the effect of recombinant ESAT-6 and CFP10 on human primary T-cell IFN-gamma secretion and proliferation. ESAT-6, but not CFP10, inhibited IFN-gamma production by T cells stimulated with M. tuberculosis or with anti-CD3 plus anti-CD28, in a dose-dependent manner. ESAT-6 also inhibited T-cell production of IL-17 and TNF-a, but not IL-2. Presence of CFP10 as part of the ESAT-6/CFP10 heterodimer did not affect ESAT-6 inhibition of T-cell IFN-gamma production. ESAT-6 inhibited the proliferation of CD3+ cells in response to TCR stimulation. ESAT-6 decreased T-cell IFN-gamma secretion by mechanisms independent of cytotoxicity or apoptosis. ESAT-6 reduced IFN-gamma mRNA levels by inhibiting the expression of the transcription factors, ATF-2, c Jun and CREB, which upregulate IFN-gamma gene expression in T cells through binding to the IFN-gamma proximal promoter. ESAT-6, but not CFP10, bound to T cells and inhibited expression of early activation markers without reducing phosphorylation of ZAP70, a proximal TCR signaling molecule. We conclude that ESAT-6 directly inhibits human T-cell responses by affecting TCR signaling pathways downstream of ZAP70. PMID- 20006312 TI - Innate and adaptive immune responses to human Mycobacterium tuberculosis infection. AB - Tuberculosis is a leading cause of death from infectious diseases world-wide, and multidrug-resistant (MDR) tuberculosis continues to spread in many parts of the world. MDR tuberculosis is a potential bioterrorist threat, as therapy is prolonged with potentially toxic agents, and the cure rate is much lower than that for treatment of drug-susceptible tuberculosis. Development of methods to enhance innate and adaptive defenses against M. tuberculosis are an attractive means to provide protection against both MDR and drug-susceptible tuberculosis. Before such strategies can be developed, an improved understanding must be gained of the immune response to M. tuberculosis. Our laboratory is mainly focused on understanding the mechanisms by which natural killer (NK) cells lyse M. tuberculosis-infected cells, determining the molecular mechanisms involved in the induction of regulatory T cells (Tregs), and characterizing the mechanisms by which NK cells affect expansion of Tregs in M. tuberculosis infection. As several studies demonstrated defective immune responses in tuberculosis patients, our studies will pinpoint the nature of this defective immune response and permit development of methods to reverse this defect. In the long run, these findings will permit development of novel methods to stimulate immunity against tuberculosis, a strategy that will contribute to development of an effective vaccine to prevent tuberculosis and novel immunotherapy to treat the disease. PMID- 20006313 TI - Pediatric rehabilitation medicine subspecialty training. PMID- 20006314 TI - The effect of running shoes on lower extremity joint torques. AB - OBJECTIVE: To determine the effect of modern-day running shoes on lower extremity joint torques during running. DESIGN: Two-condition experimental comparison. SETTING: A 3-dimensional motion analysis laboratory. PARTICIPANTS: A total of 68 healthy young adult runners (37 women) who typically run in running shoes. METHODS: All subjects ran barefoot and in the same type of stability running footwear at a controlled running speed. Three-dimensional motion capture data were collected in synchrony with ground reaction force data from an instrumented treadmill for each of the 2 conditions. MAIN OUTCOME MEASUREMENTS: Peak 3 dimensional external joint torques at the hip, knee, and ankle as calculated through a full inverse dynamic model. RESULTS: Increased joint torques at the hip, knee, and ankle were observed with running shoes compared with running barefoot. Disproportionately large increases were observed in the hip internal rotation torque and in the knee flexion and knee varus torques. An average 54% increase in the hip internal rotation torque, a 36% increase in knee flexion torque, and a 38% increase in knee varus torque were measured when running in running shoes compared with barefoot. CONCLUSIONS: The findings at the knee suggest relatively greater pressures at anatomical sites that are typically more prone to knee osteoarthritis, the medial and patellofemoral compartments. It is important to note the limitations of these findings and of current 3-dimensional gait analysis in general, that only resultant joint torques were assessed. It is unknown to what extent actual joint contact forces could be affected by compliance that a shoe might provide, a potentially valuable design characteristic that may offset the observed increases in joint torques. PMID- 20006315 TI - Adhesive capsulitis: a new management protocol to improve passive range of motion. AB - OBJECTIVE: To examine the short-term efficacy of a nonoperative shoulder protocol for the treatment of adhesive capsulitis. DESIGN: A retrospective chart review was used to collect data for a 3-year period. SETTING: Academic tertiary medical center. PATIENTS: 28 consecutive patients diagnosed as having adhesive capsulitis were identified and managed with a new protocol. METHODS: The protocol consisted of the administration of a suprascapular nerve block, the subsequent injection of an intra-articular steroid, and then the injection of an anesthetic agent with brisement normal saline volume dilation. The final step was manipulation of the shoulder. RESULTS: A paired t test was used to examine the difference in the preprocedure and postprocedure passive range of motion (flexion and abduction). The average shoulder abduction before the procedure was 89.5 degrees ; this improved by an average of 51.7 degrees (P<.0001). The average shoulder flexion improved from 117.3 degrees by an average of 37.7 degrees (P<.0001). There was no significant difference in either abduction or flexion based on age, gender, or chronicity of symptoms. White patients experienced significantly more improvement in abduction than did nonwhite patients. CONCLUSIONS: This study suggests that this adhesive capsulitis management protocol is effective and produces a significant improvement in the passive range of motion immediately after the procedure. PMID- 20006317 TI - The problem of multiple testing. PMID- 20006316 TI - Rehabilitation team disagreement: guidelines for resolution. AB - Rehabilitation care is provided by interdisciplinary teams including physicians, nurses, therapists, social workers, psychologists, and others. When there is disagreement or miscommunication among team members, progress toward patient goals can be thwarted. This paper will discuss differing models by which teams function, and what happens when a nonphysician member of the team disagrees with the plan of care. An analysis of the ethical issues embedded in two cases will be presented, and suggestions for resolution of team disagreement will be recommended. PMID- 20006318 TI - PM&R fellowship training. PMID- 20006319 TI - Use of postexercise ultrasonography to identify a symptomatic extensor digitorum longus muscle hernia associated with running. PMID- 20006320 TI - Posttraumatic osteolysis of the cervical spine mimicking a spondylodiskitis. PMID- 20006323 TI - Arguable assumptions, debatable conclusions. PMID- 20006324 TI - Regional gray matter volume in monozygotic twins concordant and discordant for schizophrenia. AB - BACKGROUND: Alterations in gray matter volume (GMV) are a robust feature of schizophrenia. However, it is not clear to what extent these abnormalities are correlates of the genetic liability to the disorder, as opposed to environmental factors and the disorder itself. We investigated the influence of genetic and environmental risk on GMV in monozygotic (MZ) twin pairs concordant and discordant for schizophrenia. METHODS: Total and regional GMVs were measured from magnetic resonance images of 80 twins: 14 MZ pairs concordant for schizophrenia, 9 pairs discordant for schizophrenia, and 17 healthy MZ twin pairs. RESULTS: Total GMV was smaller in twins with schizophrenia (t = -3.17, p = .003) and nonpsychotic cotwins from discordant pairs (t = -2.66, p = .011) than in healthy control twins. Twin pairs concordant for schizophrenia displayed reduced regional GMV in the inferior frontal, medial frontal, and anterior cingulate gyri; the caudate; lingual gyrus; and cerebellum relative to healthy twins (p < .05, corrected). Within discordant pairs, twins with schizophrenia had less GMV than their nonpsychotic cotwins in the insula; superior/medial frontal, pre/postcentral, cingulate, and superior temporal gyri; and the paracentral lobule. There were no significant differences in regional GMV between nonpsychotic cotwins and healthy control subjects. CONCLUSIONS: The presence of schizophrenia was specifically related to reduced GMV in frontal, insular, cingulate, medial parietal, and temporal cortex, over and above effects of genetic risk for the disorder. These changes could be related to the pathophysiology of the disorder itself or to unique environmental factors acting etiologically or because of the illness. PMID- 20006325 TI - The serotonin transporter promoter polymorphism is associated with cortisol response to psychosocial stress. AB - BACKGROUND: Across multiple mental health-related measures, a polymorphism (5 HTTLPR) within the promoter of the serotonin transporter gene has been associated with differential psychological sensitivity to stressful experiences. Yet, the specific mechanisms by which this polymorphism contributes to risk for psychological dysfunction is unclear. Therefore, we investigated cortisol reactivity to psychosocial stress as a potential intermediate phenotype that might predispose to such risk. METHODS: A psychologically healthy sample of 182 young adults were genotyped for the 5-HTTLPR. Each participant delivered a speech and performed mental arithmetic in one of three audience conditions: a critical evaluative audience, a supportive evaluative audience, or no audience. Salivary cortisol was sampled at baseline and at 20, 40, and 75 min after stressor onset. RESULTS: The two evaluative audience conditions elicited similar, significant increases in cortisol that were significantly greater than in the no audience control. Together, the evaluative audience conditions revealed a significant relationship between cortisol reactivity and the 5-HTTLPR, with the short/short genotype showing the greatest reactivity. Internal analyses revealed that the 5 HTTLPR was significantly associated with cortisol reactivity in the negative audience condition only, suggesting that short/short individuals might be especially vulnerable to social threat. CONCLUSIONS: The short/short genotype of the 5-HTTLPR is associated with greater cortisol reactivity to social threat. When short/short individuals experience stressful life events, they might be at greater risk for the adverse psychological and physical health consequences associated with heightened cortisol exposure. PMID- 20006327 TI - Children with Tourette's syndrome may suffer immunoglobulin A dysgammaglobulinemia: preliminary report. AB - BACKGROUND: Postinfectious autoimmunity has been implicated in Tourette's syndrome and obsessive-compulsive disorder (TS/OCD), whereas increased frequency of upper respiratory tract infections (URTI) in TS/OCD patients suggests immune deficiency. We hypothesized that antineuronal antibodies may be elevated in patients (reflecting autoimmune processes), and levels of total immunoglobulins (Igs) may be decreased (reflecting immune deficiency). METHODS: We analyzed plasma of TS/OCD patients (n = 24) and healthy age- and sex-matched control subjects (n = 22) by enzyme-linked immunosorbent assay (ELISA) for the levels of total and specific IgG, IgM, and IgA against antigens previously identified in multiple sclerosis (myelin basic protein and myelin-associated glycoprotein) and Sydenham's chorea (ganglioside-GM1, lysoganglioside, and tubulin). RESULTS: Total IgA was decreased in TS/OCD patients (median 115 mg/100 mL) compared with control subjects (141 mg/100 mL; p = .02). Specific IgA against all antigens, except tubulin were also decreased in the patients (MPB 0 vs. 13 [ELISA units [EU]; myelin-associated glycoprotein 29 vs. 44 EU, p = .04; ganglioside GM1 21 vs. 35 EU, p = .01; lysoganglioside 44 vs. 56 EU, p = .03; tubulin 44 vs. 44 EU, p = .8). The levels of total IgA and anti-myelin basic protein (MBP) IgA were significantly lower in the subgroup of pediatric autoimmune neuropsychiatric disorder associated with Streptococcus (PANDAS) cases (n = 10) than in non-PANDAS cases (n = 9; total IgA 98 mg/100 mL vs. 133 mg/mL, p = .03; anti-MBP IgA 1 vs. 6 EU, p = .03) or healthy control subjects (total IgA 141 mg/100 mL, p = .02; anti MBP IgA 13 EU, p = .005). CONCLUSIONS: At least some TS/OCD patients may suffer IgA dysgammaglobulinemia, possibly rendering the children more prone to URTI. PMID- 20006328 TI - An industrial application of a performance assessment and retuning technique for PI controllers. AB - In this paper we show how a simple methodology for the set-point following performance assessment and automatic tuning of a PI controller can be employed effectively in a real industrial application. In particular, a flow control loop in a pharmaceutical plant is considered. Practical issues related to the implementation in a Distributed Control System are discussed. Results show that the technique is capable of significantly improving the performance of the controller. PMID- 20006329 TI - Dynamic output feedback sliding mode control for uncertain mechanical systems without velocity measurements. AB - For MIMO mechanical systems using position measurements only, this paper presents a dynamic output feedback sliding mode control algorithm in which an additional dynamics is introduced into the design of the sliding surface. Although the system has the mismatched uncertainty and external disturbance, once the system is in the sliding mode, the proposed method can guarantee robust stabilization and sustain the nature of performing disturbance attenuation through utilizing H(infinity) control analytical technique. A controller is then designed to drive the system to the sliding surface in a finite time and stay on it thereafter. Finally, a numerical example is explained for demonstrating the applicability of the proposed scheme. PMID- 20006330 TI - Utility of the Oswestry Disability Index for studies of back pain related disability in nurses: evaluation of psychometric and measurement properties. AB - BACKGROUND: Disability due to back pain in nurses results in reduced productivity, work absenteeism and attrition from the nursing workforce internationally. Consistent use of outcome measures is needed in intervention studies to enable meta-analyses that determine efficacy of back pain preventive programs. OBJECTIVE: This study investigated the psychometric and measurement properties of the Oswestry Disability Index (ODI) in nursing students to determine its suitability for assessing back pain related disability in intervention studies. METHODS: Bachelor of Nursing students were recruited. Test retest reliability and the ability of the ODI to discriminate between individuals with serious and non-serious back pain were investigated. The measurement error of the ODI was examined with the minimal detectable change at the 90% confidence level (MDC(90)). RESULTS: Student nurses (n=214) had a low mean ODI score of 8.8+/-7.4%. Participants with serious back pain recorded higher scores than the rest of the cohort (p<0.05). Test-retest reliability examined in 33 individuals was ICC=0.88 (95%CI 0.77-0.94). The MDC(90)=6%, and 36% of nursing students scored below the MDC(90) indicating the tool had limited ability to detect longitudinal change in disability in this population. CONCLUSION: Data from this and previous studies demonstrate that the measurement properties of the ODI are inappropriate for studying back pain related disability in nurses. The ODI is not recommended for back pain intervention studies in the nursing population and an alternative tool that is sensitive to lower levels of disability must be determined. PMID- 20006331 TI - Thrombopoietin contributes to enhanced platelet activation in cigarette smokers. AB - OBJECTIVES: Thrombopoietin (TPO) is a humoral growth factor that primes platelet activation in response to several agonists. We recently showed that TPO enhances platelet activation in unstable angina and sepsis. Aim of this study was to investigate the role of TPO in platelet function abnormalities described in cigarette smokers. METHODS: In a case-control study we enrolled 20 healthy cigarette smokers and 20 nonsmokers, and measured TPO and C-reactive protein (CRP), as well as platelet-leukocyte binding and P-selectin expression. In vitro we evaluated the priming activity of smoker or control plasma on platelet activation, and the role of TPO in this effect. We then studied the effects of acute smoking and smoking cessation on TPO levels and platelet activation indices. RESULTS: Chronic cigarette smokers had higher circulating TPO levels than nonsmoking controls, as well as increased platelet-leukocyte binding, P selectin expression, and CRP levels. Serum cotinine concentrations correlated with TPO concentrations, platelet-monocyte aggregates and P-selectin expression. In addition, TPO levels significantly correlated with ex vivo platelet-monocyte aggregation and P-selectin expression. In vitro, the plasma from cigarette smokers, but not from nonsmoking controls, primed platelet-monocyte binding, which was reduced when an inhibitor of TPO was used. We also found that acute smoking slightly increased TPO levels, but did not affect platelet-leukocyte binding, whereas smoking cessation induced a significant decrease in both circulating TPO and platelet-leukocyte aggregation. CONCLUSION: Elevated TPO contributes to enhance platelet activation and platelet-monocyte cross-talk in cigarette smokers. PMID- 20006332 TI - Influence of menopause and cholesteryl ester transfer protein (CETP) TaqIB polymorphism on lipid profile and HDL subpopulations distribution in women with and without type 2 diabetes. AB - The common TaqIB variant in the gene coding for cholesteryl ester transfer protein (CETP), a key enzyme in reverse cholesterol transport, has been associated with higher HDL-C levels and with a more atheroprotective HDL subpopulations profile. Similar information are lacking in women with diabetes. The effect of menopause and the CETP polymorphism on lipid, lipoprotein profile, as well as on apoA-I-containing HDL subclasses distribution, as determined by two dimensional gel electrophoresis, was examined in a group of women with and without type 2 diabetes. Diabetic women showed a less atheroprotective lipid and HDL subpopulations profile, with lower levels of the large alpha-1 (P=0.006), alpha-2 (P=0.005), and prealpha-1 (P=0.02), and higher concentration of the small alpha-3 HDL particles (P=0.02) as compared to controls, independently from menopause, HDL-C and triglycerides concentrations. CETP TaqIB genotype distribution was in the Hardy-Weinberg equilibrium and comparable in the two groups, but the effect of CETP polymorphism was limited to diabetic women. In this group, the CETP variant showed significant interactions with HOMA(IR) (P<0.001), BMI (P<0.001), and triglycerides (P<0.00001), with significantly higher HDL-C levels and a more protective HDL subpopulations profile in B2 allele carriers with lower HOMA(IR), BMI or triglycerides levels. At multivariate analysis, CETP polymorphism, apoA-I, triglycerides and BMI were independent determinants of HDL-C concentration in diabetic women; apo-A-I, triglycerides, age and creatinine in controls. Type 2 diabetes is associated with a more atherogenic lipid profile; the CETP TaqIB variant may partly prevent these modifications in diabetic women with a milder degree of insulin resistance and its related disorders. PMID- 20006333 TI - The E32K variant of PCSK9 exacerbates the phenotype of familial hypercholesterolaemia by increasing PCSK9 function and concentration in the circulation. AB - OBJECTIVE: Proprotein convertase subtilisin/kexin type 9 (PCSK9) regulates cholesterol trafficking by mediating degradation of cell-surface LDL receptors (LDLR). Gain-of-function PCSK9 mutations are known to increase plasma LDL-C levels. We attempted to find gain-of-function PCSK9 mutations in Japanese subjects and determine the frequency and impacts of these mutations, especially on circulating PCSK9 and LDL-C levels. METHODS: PCR-SSCP followed by direct sequence analysis was performed for all 12 exons and intronic junctions of the PCSK9 in 55 subjects with clinically diagnosed familial hypercholesterolaemia (clinical-FH), who were confirmed to have no LDLR mutations. Among the mutations detected, PCSK9 E32K was likely to be a gain-of-function mutation, and screening was performed by PCR-RFLP in clinical-FH and general Japanese controls. The levels of PCSK9 in plasma from subjects and in media of HepG2 cells transfected with PCSK9 constructs were measured by ELISA. RESULTS: We detected 7 PCSK9 variants, including E32K. The frequency of PCSK9 E32K in clinical-FH (6.42%) was significantly higher than that in controls (1.71%). Three cases representing homozygous FH phenotypes were double heterozygous for PCSK9 E32K and LDLR C183S, C292X or K790X. Two cases were true homozygous for PCSK9 E32K; to our knowledge, these are the first true homozygotes for gain-of-function PCSK9 mutations reported to date. The PCSK9 E32K mutant had over 30% increased levels of PCSK9 in plasma from the subjects and in media of transiently transfected HepG2 cells as compared with those in controls. Furthermore, LDL-C levels in the PCSK9 E32K true homozygotes and heterozygotes were 2.10- and 1.47-fold higher than those in controls with comparable circulating PCSK9 levels, respectively, suggesting enhanced function of PCSK9 E32K. CONCLUSIONS: We found 2 true homozygotes for PCSK9 E32K and 3 double heterozygotes for PCSK9 E32K and LDLR mutations associated with autosomal dominant hypercholesterolaemia. This study provided evidence that PCSK9 E32K significantly affects LDL-C levels via increased mass and function of PCSK9, and could exacerbate the clinical phenotypes of patients carrying LDLR mutations. PMID- 20006335 TI - Aortic arch calcification detectable on chest X-ray is a strong independent predictor of cardiovascular events beyond traditional risk factors. AB - OBJECTIVES: Arterial calcification makes the management of hemodynamics more difficult. Some reports have previously shown that simple assessment of aortic calcification using plain radiography is associated with cardiovascular (CV) events; however, these studies simply assessed whether aortic calcification was present or absent only, without considering its extent. Here, we evaluated validity of grading aortic arch calcification (AAC) to predict new CV events. METHODS AND RESULTS: We retrospectively reviewed chest X-rays in 239 asymptomatic out-patients who underwent measurement of endothelial function at the 1994-2000 without past history of CV events. The extent of AAC was divided into four grades (0-3). Among these subjects, the follow-up of CV events in 209 patients was completed. At baseline, AAC grade was positively related to age, pulse pressure, diabetes and renal dysfunction. Impairment of endothelial function, as determined by flow-mediated dilation (FMD), was also correlated to increasing AAC grade. Fifty-seven CV events in total occurred during a mean follow-up period of 69+/-45 months. With multivariate adjustment, Kaplan-Meier analysis showed that the incidence was significantly higher in patients with higher AAC grade (grades 2 and 3) than in those with grade 0 or 1 (p<0.01, log-rank test). Two kinds of multivariate Cox-proportional hazards analyses showed the predictive values of AAC grade were significant (hazard ratio, 2.49; p=0.01, 2.56; p<0.01, respectively), and the predictive power was superior to that of renal dysfunction or FMD. In addition, the prediction was valuable even in patients without CKD. CONCLUSIONS: AAC detectable on chest X-ray is a strong independent predictor of CV events beyond traditional risk factors including endothelial dysfunction. Risk stratification by assessment of AAC may provide important information for management of atherosclerotic disease. PMID- 20006334 TI - High lipoprotein (a) levels are associated with an increased risk of retinal vein occlusion. AB - INTRODUCTION: Retinal vein occlusion (RVO) is one of the most common retinal vascular disorders affecting ocular vessels. Few studies, with conflicting results and conducted in limited study populations, have hypothesised the role of high levels of lipoprotein (a) [Lp(a)] in the occurrence of RVO. The aim of this study was to investigate, in a large group of RVO patients, the role of such an emerging thrombophilic parameter on the pathogenesis of RVO. MATERIALS AND METHODS: We compared 262 patients [median age: 66 years (15-88); 122 M, 140 F] with 262 age- and sex-comparable healthy subjects. RESULTS: Circulating concentrations of Lp(a) were found to be significantly different in patients when compared to healthy subjects [189 (60-1898)mg/L vs. 119.5 (6-1216)mg/L; p<0.0001, respectively]. No significant differences were observed relating to the different types of occlusion (central or branch occlusion). In order to investigate the possible association between high Lp(a) levels and the disease we performed a logistic regression analysis. In the univariate analysis, Lp(a) levels>300mg/L were found to be associated with an increased risk of RVO (OR: 2.39, 95%CI 1.39 3.59; p<0.0001). Following this, three models of multivariate analysis were performed, firstly by adjusting for age, gender, and traditional cardiovascular risk factors, secondly for triglycerides and thirdly for homocysteine levels. In all the models, Lp(a) levels>300mg/L confirmed their role as a risk factor for RVO [first model, OR: 2.15 (95%CI 1.39-3.32), p=0.0001; second model, OR: 3.11 (95%CI 1.77-5.62), p<0.00001; third model, OR: 3.48 (95%CI 1.88-6.43), p<0.00001]. CONCLUSIONS: This study reports that, in a large population of RVO patients, high Lp(a) concentrations are significantly related to RVO, independent from other traditional and emerging risk factors, suggesting that they may play a role in its pathogenesis. PMID- 20006336 TI - Using relative velocity vectors to reveal axial rotation about the medial and lateral compartment of the knee. AB - A new technique is presented that utilizes relative velocity vectors between articulating surfaces to characterize internal/external rotation of the tibio femoral joint during dynamic loading. Precise tibio-femoral motion was determined by tracking the movement of implanted tantalum beads in high-speed biplane X rays. Three-dimensional, subject-specific CT reconstructions of the femur and tibia, consisting of triangular mesh elements, were positioned in each analyzed frame. The minimum distance between subchondral bone surfaces was recorded for each mesh element comprising each bone surface, and the relative velocity between these opposing closest surface elements was determined in each frame. Internal/external rotation was visualized by superimposing tangential relative velocity vectors onto bone surfaces at each instant. Rotation about medial and lateral compartments was quantified by calculating the angle between these tangential relative vectors within each compartment. Results acquired from 68 test sessions involving 23 dogs indicated a consistent pattern of sequential rotation about the lateral condyle (approximately 60 ms after paw strike) followed by rotation about the medial condyle (approximately 100 ms after paw strike). These results imply that axial knee rotation follows a repeatable pattern within and among subjects. This pattern involves rotation about both the lateral and medial compartments. The technique described can be easily applied to study human knee internal/external rotation during a variety of activities. This information may be useful to define normal and pathologic conditions, to confirm post-surgical restoration of knee mechanics, and to design more realistic prosthetic devices. Furthermore, analysis of joint arthrokinematics, such as those described, may identify changes in joint mechanics associated with joint degeneration. PMID- 20006337 TI - Optimal handgrip height of four-wheeled walker on various road conditions to reduce muscular load for elderly users with steady walking. AB - A four-wheeled walker is a valuable tool for assisting elderly persons with walking. The handgrip height is one of the most important factor determining the usefulness of the walker. However, the optimal handgrip height for elderly users has not been considered from a biomechanical viewpoint. In this study, the handgrip height was optimized by a two-dimensional mechanical model to reduce muscular loads in the lower body as well as in the upper body with various road conditions during steady walking. A critical height of the handgrip existed at 48% of the body height for the user regardless of gender and body dimension. A lower handgrip relieved muscular load for stooping users with a lower standing height. The stooping user pushed the handgrip strongly in the perpendicular direction by leaning the upper body on the walker. However, upright users with a higher standing height should use a four-wheeled walker with a higher handgrip for maintaining his or her upright posture. For downhill movement, the optimal handgrip height depended on the slope angle and the friction coefficient between the road and the wheels of the walker. On a low-friction downhill such as asphalt with a steeper slope angle, the user was required to maintain an erect trunk with a higher handgrip and to press on the handgrip strongly in the perpendicular direction. Movement on a low-friction road was easier for users on a flat road and an uphill road, but it compelled distinct effort from users when moving downhill. PMID- 20006338 TI - Biomechanical model of human cornea based on stromal microstructure. AB - The optical characteristics of the human cornea depends on the mechanical balance between the intra-ocular pressure and intrinsic tissue stiffness. A wide range of ophthalmic surgical procedures alter corneal biomechanics to induce local or global curvature changes for the correction of visual acuity. Due to the large number of surgical interventions performed every day, a deeper understanding of corneal biomechanics is needed to improve the safety of these procedures and medical devices. The aim of this study is to propose a biomechanical model of the human cornea, based on stromal microstructure. The constitutive mechanical law includes collagen fiber distribution based on X-ray scattering analysis, collagen cross-linking, and fiber uncrimping. Our results showed that the proposed model reproduced inflation and extensiometry experimental data [Elsheikh et al., Curr. Eye Res., 2007; Elsheikh et al., Exp. Eye Res., 2008] successfully. The mechanical properties obtained for different age groups demonstrated an increase in collagen cross-linking for older specimens. In future work such a model could be used to simulate non-symmetric interventions, and provide better surgical planning. PMID- 20006339 TI - Cells in 3D matrices under interstitial flow: effects of extracellular matrix alignment on cell shear stress and drag forces. AB - Interstitial flow is an important regulator of various cell behaviors both in vitro and in vivo, yet the forces that fluid flow imposes on cells embedded in a 3D extracellular matrix (ECM), and the effects of matrix architecture on those forces, are not well understood. Here, we demonstrate how fiber alignment can affect the shear and pressure forces on the cell and ECM. Using computational fluid dynamics simulations, we show that while the solutions of the Brinkman equation accurately estimate the average fluid shear stress and the drag forces on a cell within a 3D fibrous medium, the distribution of shear stress on the cellular surface as well as the peak shear stresses remain intimately related to the pericellular fiber architecture and cannot be estimated using bulk-averaged properties. We demonstrate that perpendicular fiber alignment of the ECM yields lower shear stress and pressure forces on the cells and higher stresses on the ECM, leading to decreased permeability, while parallel fiber alignment leads to higher stresses on cells and increased permeability, as compared to a cubic lattice arrangement. The Spielman-Goren permeability relationships for fibrous media agreed well with CFD simulations of flow with explicitly considered fibers. These results suggest that the experimentally observed active remodeling of ECM fibers by fibroblasts under interstitial flow to a perpendicular alignment could serve to decrease the shear and drag forces on the cell. PMID- 20006341 TI - Identification and enantioselective gas chromatographic mass-spectrometric separation of O-desmethylnaproxen, the main metabolite of the drug naproxen, as a new environmental contaminant. AB - O-desmethylnaproxen (2-(6-hydroxynaphthalen-2-yl)propanoic acid) was identified in 10 different water samples from Germany and Pakistan. In the Pakistan samples it was found in all samples, surface water and effluents, exhibiting estimated concentrations between 0.04 and 1.36 microg/L. In Germany it was only encountered in the STP-effluent with an average concentration of 0.23 microg/L. Furthermore, enantioselective GC analyses revealed differences in the enantiomeric ratios found in Germany and Pakistan. To the best of our knowledge this is the first report on the identification of O-desmethylnaproxen, the main metabolite of the drug naproxen, in environmental samples. PMID- 20006342 TI - Automated determination of aliphatic primary amines in wastewater by simultaneous derivatization and headspace solid-phase microextraction followed by gas chromatography-tandem mass spectrometry. AB - This paper presents a fully automated method for determining ten primary amines in wastewater at ng/L levels. The method is based on simultaneous derivatization with pentafluorobenzaldehyde (PFBAY) and headspace solid-phase microextraction (HS-SPME) followed by gas chromatography coupled to ion trap tandem mass spectrometry (GC-IT-MS-MS). The influence of main factors on the efficiency of derivatization and of HS-SPME is described in detail and optimized by a central composite design. For all species, the highest enrichment factors were achieved using a 85 microm polyacrylate (PA) fiber exposed in the headspace of stirred water samples (750 rpm) at pH 12, containing 360 g/L of NaCl, at 40 degrees C for 15 min. Under optimized conditions, the proposed method achieved detection limits ranging from 10 to 100 ng/L (except for cyclohexylamine). The optimized method was then used to determine the presence of primary amines in various types of wastewater samples, such as influent and effluent wastewater from municipal and industrial wastewater treatment plants (WWTPs) and a potable water treatment plant. Although the analysis of these samples revealed the presence of up to 1500 microg/L of certain primary amines in influent industrial wastewater, the concentration of these compounds in the effluent and in municipal and potable water was substantially lower, at low microg/L levels. The new derivatization-HS SPME-GC-IT-MS-MS method is suitable for the fast, reliable and inexpensive determination of primary amines in wastewater in an automated procedure. PMID- 20006340 TI - In vitro stable isotope labeling for discovery of novel metabolites by liquid chromatography-mass spectrometry: Confirmation of gamma-tocopherol metabolism in human A549 cell. AB - A general approach for discovering novel catabolic metabolites from a parent biocompound was developed and validated on the metabolism of gamma-tocopherol in human A549 cell. The method is based on LC-MS analysis of in vitro stable isotope labeled metabolites and assumes that a parent compound and its metabolites share a common functional group that can be derivatized by well-documented reagents. In this method, two equal aliquots of extracted metabolites are separately derivatized with isotope-coded (heavy) and non-isotope-coded (light) form of derivatizing reagent, mixed at 1:1 ratio and analyzed using LC-MS. The metabolites with common functional group are then easily recognized by determination of a chromatographically co-eluted pair of isotopomers (MS doublet peaks) with similar peak intensities and mass difference corresponding to the mass difference between heavy and light form of derivatization reagent. The feasibility of this approach was demonstrated and validated by the identification of products of gamma-tocopherol catabolism in human A549 cell culture media using N-methyl-nicotinic acid N-hydroxysuccinimide ester (C1-NANHS) and N-methyl-d3 nicotinic acid N-hydroxysuccinimide ester (C1-d3-NANHS) derivatizing reagent. Overall four gamma-tocopherol metabolites were identified including 9'-COOH, 11' COOH, 13'-COOH and 13'-OH. In addition, the developed LC-MS method can also be used for the fast and sensitive quantitative analysis of gamma-tocopherol and other forms of vitamin E related compounds. PMID- 20006343 TI - Adsorption and surface complex modeling of silicates on maghemite in aqueous suspensions. AB - Adsorption of soluble silicate species (Si(OH)(4)) on maghemite (gamma-Fe(2)O(3)) has been studied by high precision potentiometric titrations, spectroscopic analyses, and zeta potential determinations. Titrations were performed at 25.0 degrees C within the range 2.8FeOH(2)(+); logbeta(11)(s)(int)=5.39+/-0.01FeOH<-->FeO(-)+H(+); logbeta(-11)(s)(int)=-7.51+/ 0.01; C=2.40Fm(-2). Three different concentration ratios of the determined concentration of active surface sites and added total silicate concentration [FeOH]:[Si(OH)(4)] were examined (1:1,2:1, and 3:1). A model comprising three surface complexes, FeOSi(OH)(3),logbeta(011)(s)(int)=3.61+/-0.02; FeOSiO(OH)(2)( ),logbeta(111)(s)(int)=-3.00+/-0.01; and FeOSiO(2)(OH)(2-),logbeta(-211)(s)(int)= 11.35+/-0.02, was found to best describe the experimental observations. Attempts to model the adsorption of silicates on maghemite as bidentate or polynuclear silicate complexes were not successful. The maximum silicate adsorption for the 1:1 ratio, approximately 80%, was obtained at pH 9-9.5. The IEP of maghemite in the presence of silicates (1:1 ratio) was determined from zeta-potential measurements, giving pH(IEP) approximately 6.6. PMID- 20006345 TI - The operating room dashboard. PMID- 20006344 TI - ADHD familial loading and abnormal EEG alpha asymmetry in children with ADHD. AB - OBJECTIVE: Abnormal brain laterality (ABL) is indicated in ADHD. ADHD and brain laterality are heritable. Genetic factors contributing to lateralization of brain function may contribute to ADHD. If so, increased ADHD family loading should be associated with greater ABL. Previous studies have shown increased rightward alpha asymmetry in ADHD. We tested whether this was more pronounced in ADHD children with increased ADHD family loading. METHODS: We compared EEG alpha asymmetry at rest and during the Conner's Continuous Performance Test (CPT) in ADHD children with and without ADHD affected parents, and replicated our findings in a second larger sample. The replication study additionally stratified the parent-affected sample by parental persistent versus non-persistent ADHD status, increased spatial resolution of EEG measures, and assessed low versus high-alpha. RESULTS: Study-1: the parent-affected group showed increased rightward asymmetry across frontal and central regions and reduced rightward parietal asymmetry during an eyes closed (EC) condition, as well as increasing rightward parietal asymmetry with advancing age during the CPT. Study-2 replicated these findings and further delineated influences of low versus high-alpha, recording site, and effects of parental persistent versus non-persistent ADHD status. CONCLUSION: Increased ADHD familial loading was associated with increased rightward frontal asymmetry. In contrast, increased rightward parietal asymmetry was associated with reduced ADHD family loading. Frontal results are consistent with an ADHD endophenotype. Parietal results suggest an ADHD adaptive trait prevalent with less ADHD family loading. Age effects indicate a unique developmental course among ADHD children whose parents have non-persistent ADHD. PMID- 20006346 TI - Postoperative hypoxemia: common, undetected, and unsuspected after bariatric surgery. AB - BACKGROUND: Patients undergoing gastric bypass are at greater than ordinary risk for postoperative respiratory insufficiency, presumably related to obstructive sleep apnea (OSA) and patient-controlled analgesia (PCA). This study was proposed to quantify the magnitude of the problem. METHODS: Fifteen patients undergoing gastric bypass had oxygen saturation (SpO(2)) recorded continuously, but not displayed, for 24h postoperatively; eight also had arterial blood analysis every 4h. All received narcotic PCA. SpO(2)<90% lasting more than 10 s was reviewed. Results are mean+/-SEM. RESULTS: Mean age was 44+/-4 y, and mean BMI was 48+/ 2kg/m(2); 77% had OSA. Every patient had more than one episode with SpO(2)<90% for longer than 30s undetected by routine monitoring; most had multiple episodes. Nadir SpO(2) averaged 75% +/- 8%. Mean longest duration of desaturation below 90% averaged 21+/-15min. Mean PaCO(2) was 37+/-3mm Hg; maximum PaCO(2) was 47mm Hg. CONCLUSIONS: Severe and prolonged episodes of hypoxemia were a consistent finding, despite aggressive preoperative diagnosis and treatment of OSA, including use of CPAP postoperatively. Although some postoperative hypoventilation was expected, the degree and frequency of desaturation were surprising. No patient exhibited arterial PaCO(2) evidence of hypoventilation. No patient experienced cardiopulmonary arrest/instability, in spite of severe, repeated episodes of hypoxemia. In no instance was a significant hypoxemic episode suspected or detected. Continuous pulse oximetry monitoring, with an audible alarm set for a saturation less than 90% for 10 s, would have alerted providers to 100% of significant hypoxemic episodes. Our recommendation is routinely monitoring (with alarm capability enabled) every bariatric surgical patient, to prevent such occurrence. PMID- 20006347 TI - The potential effect of proteasome inhibitor PS-341 on severe acute pancreatitis detected by positron emission tomography in ICR mice. AB - INTRODUCTION: Severe acute pancreatitis is a life threatening disease with a high rate of mortality, and its treatments are still controversial. The purpose of this study is to investigate the potential effects of proteasome inhibitor PS-341 on severe acute pancreatitis induced by cerulein and lipopolysaccharide in mice. MATERIALS AND METHODS: Severe acute pancreatitis was induced by seven intraperitoneal injections of 50 ug/kg cerulein at hourly intervals and one injection of 10mg/kg lipopolysaccharide in mice. Thirty min before the administration of lipopolysaccharide, mice were treated either with PS-341 or vehicle. The severity of acute pancreatitis was then evaluated by serum and pancreatic biochemical assays as well as histologic examination. Positron emission tomography (PET) was used for the first time to determine the therapeutic effects of interventions in situ. RESULTS: PS-341 significantly inhibited NF-kappaB activation, while the pancreatic cell apoptosis was significantly enhanced, resulting in the improved parameters such as serum amylase, C-reactive protein, lactate dehydrogenase, interleukin-1beta, interleukin-6, and pancreatic myeloperoxidase activity. Accordingly, pancreatic damage, including inflammatory cell infiltration, hemorrhage, and necrosis, was markedly reduced. (18)F-fluorodeoxyglucose-positron emission tomography demonstrated that PS-341 significantly reduced the uptake of (18)F fluorodeoxyglucose within the pancreas. CONCLUSIONS: These observations demonstrate that PS-341 was able to significantly reduce the severity of acute pancreatitis induced by cerulein and lipopolysaccharide in mice. The potential effect is associated with the inhibition of NF-kappaB activation and increased pancreatic cell apoptosis within the pancreas. (18)F-fluorodeoxyglucose-positron emission tomography could be a sensitive and promising means in evaluating the therapeutic effect and adjusting medical interventions for pancreatitis. PMID- 20006348 TI - Bosentan affects 15-F2t-isoprostane adverse effects on postischemic rat hearts. AB - BACKGROUND: 15-F(2t)-isoprostane (IsoP), a marker of reactive oxygen species induced oxidative stress, is increased after myocardial ischemia and reperfusion. It exerts deleterious effects on postischemic myocardium accompanied with increased release of endothelin-1 (ET-1), a potent vasoconstrictor. We hypothesized that IsoP exacerbates myocardial ischemia-reperfusion injury by stimulating ET-1 production, and that ET-1 blockade can attenuate or prevent these deleterious effects of IsoP. METHODS: Adult rat hearts were perfused by the Langendorff technique with Krebs-Henseleit solution (KH) at a constant flow rate of 10 mL/min. Global myocardial ischemia was induced by stopping KH perfusion for 40 min followed by 60 min of reperfusion. Hearts were randomized to one of the five groups (n = 8 each): untreated control, treated with IsoP (100 nM), or the ET-1 receptor A/B antagonist bosentan (1 MUM) alone or in combination 10 min prior to, during 40 min global ischemia and 15 min of reperfusion, or treated with IsoP as above plus delayed administration of bosentan after 15 min of reperfusion. RESULTS: Coronary effluent ET-1 concentrations in the IsoP group were higher than those in the control group during ischemia and reperfusion (P < 0.05), which was associated with increased release of cardiac-specific creatine kinase, reduced cardiac contractility during reperfusion, and increased myocardial infarct size (all P < 0.05 versus control). Bosentan administration during early reperfusion exacerbated the IsoP deleterious effects, while delayed administration attenuated it. CONCLUSION: 15-F(2t)-isoprostane-induced ET-1 production during later reperfusion is detrimental to functional recovery of damaged myocardium, while ET-1 increase during early reperfusion seems to improve it. PMID- 20006349 TI - Traumatic brain injury and aeromedical evacuation: when is the brain fit to fly? AB - BACKGROUND: To review the inflammatory sequelae of traumatic brain injury (TBI) and altitude exposure and discuss the potential impact of aeromedical evacuation (AE) on this process. METHODS: Literature review and expert opinion regarding the inflammatory effects of TBI and AE. RESULTS: Traumatic brain injury has been called the signature injury of the current military conflict. As a result of the increasing incidence of blast injury, TBI is responsible for significant mortality and enduring morbidity in injured soldiers. Common secondary insults resulting from post-traumatic cerebral inflammation are recognized to adversely impact outcome. AE utilizing Critical Care Air Transport Teams has become a standard of care practice following battlefield injury, to quickly and safely transport critically injured soldiers to more sophisticated echelons of care. Exposure to the hypobaric conditions of the AE process may impose an additional physiologic risk on the TBI patient as well as a "second hit" inflammatory stimulus. CONCLUSIONS: We review the known inflammatory effects of TBI and altitude exposure and propose that optimizing the post-traumatic inflammatory profile may assist in determining an ideal time to fly for head-injured soldiers. PMID- 20006350 TI - Rectus sheath pressure as a novel diagnostic adjunct for abdominal compartment syndrome. PMID- 20006351 TI - Limitations of the SEER database for demonstrating causal relationships between treatments and outcomes in pediatric intestinal tumors. PMID- 20006353 TI - Facial emotion recognition and cerebral white matter lesions in myotonic dystrophy type 1. AB - In order to investigate the cognitive and neurological bases of social cognitive impairment in myotonic dystrophy type 1 (DM1), we examined the facial expression recognition abilities and the cerebral lesions in a group of DM 1 (5 men, 4 women). We measured sensitivity to facial emotions and compared the findings with magnetic resonance image (MRI) findings evaluated using a semi-quantitative method. The DM1 patients showed lower sensitivity to disgusted and angry faces as compared to the healthy controls. The assessment of brain lesions revealed that more severe lesions occurred in the frontal, temporal, and insular white matters. Sensitivity to the emotion of disgust was negatively correlated with temporal lesions, and sensitivity to anger negatively correlated with frontal, temporal, and insular lesions. The results of this study indicate an association between lesions in the frontal, temporal, and insular subcortices and decreased emotional sensitivity to disgust and anger in DM1 patients. These areas are thought to play an important role in emotional processing in the normal brain. Our results suggest that social cognitive impairment in DM1 patients is attributable to impaired emotional processing linked to white matter lesions. PMID- 20006352 TI - The effects of exogenous melatonin on peripheral nerve regeneration and collagen formation in rats. AB - BACKGROUND: Peripheral nerve damage that requires surgical repair does not result in complete recovery because of collagen scar formation, ischemia, free oxygen radical damage, and other factors. To date, the best treatment method has not yet been determined. In this study, we designed an experimental peripheral nerve injury model, and researched the possible effects of melatonin hormone, based on evidence of its strong antioxidant and cell-protective effects via mimicking the effects of calcium channel blockers. MATERIALS AND METHODS: We randomized 24 healthy female albino rats into three groups: the pinealectomy group, melatonin group, and control group. In the pinealectomy group, craniotomy, pinealectomy, sciatic nerve transection, and coaptation were performed, and 0.9% NaCl was injected intraperitoneally. In the melatonin group, craniotomy (without pinealectomy), sciatic nerve dissection, and coaptation were performed, and melatonin was injected intraperitoneally, instead of NaCl. In the control group, craniotomy (without pinealectomy), sciatic nerve dissection and coaptation, and intraperitoneal NaCl injection were performed. In each group, nerve recovery was evaluated histologically, functionally, and electrophysiologically. Functional and electrophysiologic evaluations were conducted before surgery and at 4 and 12 wk. RESULTS: At 4 wk, no significant difference was observed between the groups. However, at 12 wk, significant electrophysiologic and functional improvement was observed only in the melatonin group. CONCLUSIONS: Melatonin seems to have a beneficial effect on nerve recovery. However, this effect is not effective at physiologic doses. Future comparative studies with melatonin versus other nerve regenerating agents are necessary to determine the clinical utility of melatonin hormone. PMID- 20006355 TI - Predictors of survival in patients with persistent nodal metastases after preoperative chemotherapy for esophageal cancer. AB - OBJECTIVE: In patients with esophageal cancer, a complete pathologic response after preoperative therapy is universally regarded as a favorable prognostic factor. However, less is known about factors predictive of outcome in patients with persistent nodal disease. The purpose of this study is to determine which variables affect survival in this patient population. METHODS: We reviewed a prospectively maintained esophageal cancer database. Patients with positive lymph nodes after preoperative therapy and surgery were selected. Predictors of survival were examined univariately using the log-rank test. Factors identified at P < .20 by univariate analysis were selected for inclusion in a multivariate model. RESULTS: Ninety-six patients with 1 or more positive nodes received preoperative therapy. Pathologic T classification was 0 to 2 in 25 (26%) patients and 3 to 4 in 71 (74%) patients. In 29 (30%) patients, nonregional nodal disease was present (M1). Final pathologic stages were IIB in 18 (19%), III in 49 (51%), and IV in 29 (30%). Postoperatively, 44 (46%) patients received additional chemotherapy. On univariate analysis, pathologic stage, pathologic T classification, and number of positive nodes significantly affected overall survival. On multivariate analysis, clinical stage (hazard ratio [HR], 2.25; P = .05), pathologic T classification (HR, 3.06; P = .006), and number of positive nodes (HR 1.03 per node, P = .09) were significant predictors of overall survival. CONCLUSION: Long-term survival can be achieved in patients with esophageal cancer who have persistent nodal disease after neoadjuvant therapy and surgical resection. Clinical stage, pathologic T classification, and number of positive nodes best predict survival. Nonregional nodal disease does not adversely affect outcome. Postoperative chemotherapy conferred no additional survival benefit in this patient population. PMID- 20006356 TI - The Graft Imaging to Improve Patency (GRIIP) clinical trial results. AB - OBJECTIVE: This trial aimed to determine whether intraoperative graft assessment with criteria for graft revision would decrease the proportion of patients with 1 or more graft occlusions or stenoses or major adverse cardiac events 1 year after coronary artery bypass grafting. METHODS: A single-center, randomized, single blinded, controlled clinical trial was designed. Patients were randomized to either of 2 groups: intraoperative graft patency assessment using indocyanine green fluorescent angiography and transit-time flowmetry, with graft revision according to a priori criteria (imaging group), or standard intraoperative management (control group). Patients underwent follow-up angiography at 1 year. RESULTS: Between September 2005 and August 2008, 156 patients undergoing isolated coronary bypass grafting were enrolled (imaging, n = 78; control, n = 78). Demographic and angiographic characteristics were similar between groups. Operative, crossclamp, and cardiopulmonary bypass times were all nonsignificantly longer in the imaging arm. The number of grafts per patients was similar (imaging, 3.0 +/- 0.7; control, 3.0 +/- 0.7). The frequency of major adverse cardiac events (death, myocardial infarction, repeat revascularization) was not different between groups at 1 year postoperatively (imaging, 7.7%; control, 7.7%). One-year angiography was performed in 107 patients (imaging, 55 patients/160 grafts; control, 52 patients/152 grafts). The proportion of patients with 1 graft occlusion or more was comparable in the imaging (30.9%) and control (28.9%) groups (relative risk [95% confidence interval], 1.1 [0.6-1.9]; P = .82), as were other graft patency end points. The incidence of saphenous vein graft occlusion was high in both groups. CONCLUSIONS: Routine intraoperative graft assessment is safe but does not lead to a marked reduction in graft occlusion 1 year after bypass grafting. The incidence of saphenous vein graft failure remains high despite contemporary practice and routine intraoperative graft surveillance. PMID- 20006357 TI - Outcomes of reoperative aortic valve replacement after previous sternotomy. AB - OBJECTIVE: Increasingly, patients with previous sternotomy require aortic valve replacement. We compared outcomes of reoperative aortic valve replacement after previous sternotomy and primary aortic valve replacement by surgical era. Effect of initial cardiac operation on reoperative aortic valve replacement was also investigated. METHODS: Between January 1996 and December 2007, a total of 1603 patients undergoing elective aortic valve replacement were entered prospectively into our clinical database. Patients were divided into eras A (1996-1999), B (2000-2003), and C (2004-2007). A total of 191 patients (12%) had previous sternotomy for coronary artery bypass grafting (n = 88), coronary artery bypass grafting with aortic valve replacement (n = 16), aortic valve replacement with or without other aortic procedure (n = 70), and other cardiac procedures (n = 17). Mean ages were 66.5 +/- 13.1 years in reoperative group and 65.5 +/- 14.9 years in primary group. RESULTS: Mortality in reoperative group decreased significantly with time (A 15.4% vs B 15.1% vs C 2.0%, P = .004) and was equivalent to primary group in era C (3.5% vs 2.0%, P = .65). Major complications also significantly decreased with time in reoperative group (A 25.6% vs B 17.0% vs C 6.1%, P = .006). Importantly, patients had more comorbidities with time and increased preoperative risk in era C. There were no differences in outcome by initial cardiac operation in reoperative group. CONCLUSIONS: Reoperative aortic valve replacement now carries similar morbidity and mortality to primary replacement. Risk of reoperation is not affected by primary operation. PMID- 20006360 TI - Analysis of foraminifera assemblages and sediment geochemical properties to characterise the environment near Araca and Saco da Capela domestic sewage submarine outfalls of Sao Sebastiao Channel, Sao Paulo State, Brazil. AB - Superficial bottom samples were collected near diffusers of domestic sewage submarine outfalls at Araca and Saco da Capela, Sao Sebastiao Channel, Brazil. The goal of this study was to investigate the distribution and composition of live benthic foraminifera assemblages and integrate the results obtained with geochemical analyses to assess human-induced changes. According to the results obtained no environmental stress was observed near the Saco da Capela submarine outfall diffusers. The foraminifera assemblage is characterised by species typical of highly hydrodynamic environments, with well-oxygenated bottom waters and low nutrient contents. In contrast, near Araca submarine outfall, organic enrichment was denoted by high phosphorus, sulphur and, to a lesser extent, total organic carbon content. Harmful influences on foraminifera could be identified by low richness and specific diversity, as well as the predominance of detritivore feeder species, which are associated with higher organic matter flux and low oxygen in the interstitial pore water. PMID- 20006361 TI - HELCOM Baltic Sea Action Plan--a regional programme of measures for the marine environment based on the Ecosystem Approach. AB - The Helsinki Commission (HELCOM) Baltic Sea Action Plan, adopted by the coastal countries of the Baltic Sea and the European Community in November 2007, is a regional intergovernmental programme of measures for the protection and management of the marine environment explicitly based on the Ecosystem Approach. The Action Plan is structured around a set of Ecological Objectives used to define indicators and targets, including effect-based nutrient input ceilings, and to monitor implementation. The Action Plan strongly links Baltic marine environmental concerns to important socio-economic fields such as agriculture and fisheries and promotes cross-sectoral tools including marine spatial planning. Due to complementarities with the European Union (EU) Marine Strategy Framework Directive, the Action Plan is in essence a pilot for this process without neglecting the important role of the Russian Federation - the only Baltic coastal country not a member of the EU. PMID- 20006362 TI - Enhanced levels of soluble CD40 ligand and C-reactive protein in a total of 312 patients with metabolic syndrome. AB - The metabolic syndrome (MS) is associated with a systemic inflammatory response that plays an important pathogenetic role in atherothrombotic disease. Increasing evidence indicates that CD40-CD40 ligand interactions constitute an important mediator for vascular inflammation. The purpose of this study was to assess whether high-sensitivity C-reactive protein (hs-CRP) and soluble CD40 ligand (sCD40L) levels were increased in patients with MS. During the study period from January 2004 to August 2004, 312 patients with MS and 98 control subjects were included. Anthropometric measurements, blood pressure assessment, electrocardiography, and blood measurements including fasting blood glucose, postprandial blood glucose, total cholesterol, low-density lipoprotein cholesterol, high-density lipoprotein cholesterol, triglyceride, glycated hemoglobin, white blood cell (WBC), platelets, hs-CRP, and sCD40L were performed. Patients with MS were divided into 3 groups based upon their glucose tolerance (group 1, normal glucose tolerance; group 2, prediabetic group; and group 3, diabetes mellitus). Patients with MS showed a significant increase of WBC, hs CRP, and sCD40L levels compared with control subjects. The levels of both hs-CRP and sCD40L were positively correlated with body mass index (BMI). High sensitivity CRP levels were also positively correlated with waist circumferences, fasting blood glucose, postprandial blood glucose, and glycated hemoglobin, and negatively correlated with high-density lipoprotein cholesterol. In patients with MS, both hs-CRP and sCD40L levels were positively correlated with WBC count. We found a positive correlation between sCD40L and platelets. Among the subgroups of patients with MS, the mean levels of WBC, hs-CRP, and sCD40L did not show any significant differences. In conclusion, elevated levels of WBC, hs-CRP, and sCD40L in MS patients provide further insight into the relationship between MS and inflammation. In our study, positive correlations between BMI and both hs-CRP and sCD40L levels suggest that BMI is an important determinant of a chronic inflammatory state in patients with MS. Moreover, this study reports significantly increased levels of WBC, hs-CRP, and sCD40L not only in diabetic subjects with MS but also in prediabetic subjects and nondiabetic subjects with MS compared with control subjects. Our data suggest that MS patients have proinflammatory state independent of their glucose tolerance status. In our study, the positive correlation between the levels of sCD40L and platelets in patients with MS supports previous reports indicating that sCD40L are derived predominantly from platelets. PMID- 20006363 TI - Baseline forced expiratory volume in the first second as an independent predictor of development of the metabolic syndrome. AB - A growing body of evidence strongly supports associations between reduced lung function and insulin resistance, type 2 diabetes mellitus, and cardiovascular disease. The present study was undertaken to explore the possibility that reduced lung function is an independent predictor of development of the metabolic syndrome (MetS) and to investigate potential links between reduced lung function and the MetS. A prospective cohort study of reduced lung function as a predictor of subsequent MetS was conducted using 2-year follow-up data for 450 middle-aged adults lacking the MetS at baseline. Data were obtained from the Taipei MJ Health Screening Centers in Taiwan. The MetS was defined according to the modified Adult Treatment Panel III criteria. Over 2 years of follow-up, 26 of the 450 subjects (5.78%) without the MetS at baseline subsequently developed the syndrome. In multiple logistic regression analysis with adjustments for age, sex, body mass index, cigarette smoking, alcohol consumption, and physical activities, reduced forced expiratory volume in the first second (FEV(1)) at baseline remained a predictor of subsequent MetS (relative risk of 4.644, P = .036 for the third [<2.31 L] vs first [>2.88 L] tertile). In Pearson and partial correlation analyses, white blood cell counts and C-reactive protein concentrations were both found to be significantly and negatively correlated with FEV(1). Lower FEV(1) is concluded to serve as an independent predictor of the MetS. Low-grade systemic inflammation is the possible link between reduced lung function and the MetS. PMID- 20006364 TI - Palmitate induces insulin resistance without significant intracellular triglyceride accumulation in HepG2 cells. AB - Previous studies showed that increased release of free fatty acids from adipocytes leads to insulin resistance and triglyceride (TG) accumulation in the liver, which may progress into hepatic steatohepatitis. We and other investigators have previously reported that palmitate induces endoplasmic reticulum stress-mediated toxicity in several tissues. This work investigated whether palmitate could induce insulin resistance and steatosis in HepG2 cells. We treated cells with either saturated fatty acid (palmitate) or unsaturated fatty acid (oleate), and observed that palmitate significantly activated c-jun N terminal kinase and inactivated protein kinase B. Both 4-phenylbutyric acid and glycerol significantly activated protein kinase B, confirming the involvement of endoplasmic reticulum stress in palmitate-mediated insulin resistance. Oleate, but not palmitate, significantly induced intracellular TG deposition and activated sterol regulatory element binding protein-1. Instead, diacylglycerol level and protein kinase C epsilon activity were significantly increased by palmitate, suggesting the possible role of diacylglycerol in palmitate-mediated lipotoxicity. Therefore, the present study clearly showed that palmitate impairs insulin resistance, but does not induce significant TG accumulation in HepG2 cells. PMID- 20006365 TI - Biological markers, lifestyle factors, and metabolic syndrome. PMID- 20006366 TI - Cardiovascular protective flavonolignans and flavonoids from Calamus quiquesetinervius. AB - Tricin-type flavonolignans, (2S)-dihydrotricin 4'-O-(erythro-beta guaiacylglyceryl) ether, (2S)-dihydrotricin 4'-O-(threo-beta-guaiacylglyceryl) ether, (2S)-dihydrotricin 4'-O-(threo-beta-4-hydroxyphenylglyceryl) ether, tricin 4'-O-(erythro-beta-4-hydroxyphenylglyceryl) ether, tricin 4'-O-(threo-beta-4 hydroxylphenylglyceryl) ether, and (2S)-dihydrotricin 4'-O-(beta-6''-methoxy-4'' oxo-chroman-3''-yloxy) ether namely calquiquelignan A-F, respectively, were isolated and characterized from the EtOAc extract of Calamus quiquesetinervius. Additionally, six known phenolic compounds, including dihydrotricin, tricin, salcolin A, p-hydroxybenzoic acid, (2S, 3S)-trans-dihydrokapempferol and (2S) naringenin, were also obtained and identified from the extract. Structures of the flavonolignans were assigned based on spectroscopic analyses that included 1D and 2D NMR spectroscopic techniques, such as HMQC, HMBC, and NOESY. Bioassay results showed that calquiquelignan A, dihydrotricin and (2S)-naringenin exhibited significant vasodilatory potencies, as indicated by 60.3%, 80.3% and 60.9% relaxations, respectively, at 100 microM. Salcolin A showed potent platelet aggregation inhibition, compared with aspirin. Most of the tricin-type derivatives (calquiquelignan A-B, dihydrotricin and tricin) also exhibited more potent hydroxyl radical ((.)OH) scavenging activity than trolox as characterized by the ultraweak chemiluminescence assay. PMID- 20006368 TI - Mycoreovirus 1 S4-coded protein is dispensable for viral replication but necessary for efficient vertical transmission and normal symptom induction. AB - Rearrangements of two segments, S6 and S10, of Mycoreovirus 1 (MyRV1), a member of the family Reoviridae, were previously shown to be induced at a high rate by the multifunctional protein p29 encoded by a distinct ssRNA virus, the prototype hypovirus CHV1-EP713 (Sun and Suzuki, RNA 14, 2557-2571, 2008). Here we report the occurrence of rearrangements of MyRV1 S4, albeit at a very low frequency, in the absence of CHV1 p29, resulting in internal 80-90% deletions of the open reading frame (ORF) in S4. Comparative analyses of fungal strains infected by wild-type MyRV1 and its variants carrying rearrangements of S4, S4 plus S10 and S10 indicated that S4-encoded VP4, like VP10, is non-essential for virus replication but required for efficient vertical transmission and symptom expression caused by MyRV1. This is the first example of a reovirus variant that carries deletions of over 75% of the ORFs in two genome segments and is still replication-competent. PMID- 20006367 TI - Binding of RBP-Jkappa (CSL) protein to the promoter of the Kaposi's sarcoma associated herpesvirus ORF47 (gL) gene is a critical but not sufficient determinant of transactivation by ORF50 protein. AB - ORF50 protein activates the KSHV lytic cycle. The promoter of an early lytic cycle gene ORF47, encoding envelope protein gL, is activated by an interaction between ORF50 protein and RBP-Jkappa. In ORF47p only one of two sequences fitting the consensus RBP-Jkappa recognition site strongly binds RBP-Jkappa and confers a response to ORF50 protein. Flanking sequences 5' to the RBP-Jkappa binding site are required to confer a maximal response to ORF50 protein. Not all mutant ORF50 response elements in the ORF47p that are bound by RBP-Jkappa are sufficient to confer maximal ORF50 responsiveness. Four sequences containing an RBP-Jkappa site and flanking sequences characteristic of the ORF50 response element in ORF47p were identified in human DNA. All bound RBP-Jkappa, but only one responded robustly to ORF50 protein. We propose models for the possible function of ancillary sequences flanking the RBP-Jkappa-binding element which are crucial for mediating ORF50 transactivation. PMID- 20006369 TI - Site-specific attachment of palmitate or stearate to cytoplasmic versus transmembrane cysteines is a common feature of viral spike proteins. AB - Many glycoproteins of enveloped viruses are known to be "palmitoylated" at cysteines located either in the transmembrane region or in the cytoplasmic tail. Although it was recognized early on that "palmitoylation" is not specific for this carbon chain, the exact fatty acid composition of S-acylated proteins has been difficult to determine. Advancements in mass-spectrometry (MS) now allow one to quantify the fatty acids linked to single acylation sites. We report that G of Vesicular Stomatitis virus contains palmitate at a cytoplasmic cysteine, whereas F of Newcastle Disease virus and E1 of Semliki Forest virus (SFV) are stoichiometrically acylated with stearate at a transmembrane cysteine. E2 of SFV contains three molecules of palmitate and one molecule of stearate, the latter probably attached to a transmembrane cysteine. Thus, site-specific attachment of palmitate or stearate, previously described only for HA of influenza virus, is a common feature of viral spike proteins. PMID- 20006370 TI - Systematic identification of microRNA and messenger RNA profiles in hepatitis C virus-infected human hepatoma cells. AB - In order to investigate the global and dynamic host microRNAs (miRNAs)/messenger RNAs (mRNAs) expression alteration during in vitro acute HCV infection, a comprehensive microarray analysis was performed using human hepatoma cells. Totally, 108 human miRNAs and 1247 mRNAs were identified whose expression levels changed for more than 2.0-fold in response to HCV infection. Upon HCV infection, signature from the unique miRNA expression pattern reflected the involvement of miRNA-regulated host cellular physiology and antiviral mechanism, whereas a preponderance of differentially regulated genes associated with metabolism, cell growth, apoptosis and cytokine/chemokine pathways. Furthermore, a reverse regulatory association of differentially expressed miRNAs and their predicted targets was constructed. Finally, the differentially expressed miRNAs such as miR 24, miR-149, miR-638 and miR-1181 were identified to be involved in HCV entry, replication and propagation. These results suggest that combined miRNA and mRNA profiling may have superior potential as a diagnostic and mechanistic feature in HCV infection. PMID- 20006371 TI - Temporal trends of triclosan contamination in dated sediment cores from four urbanized estuaries: evidence of preservation and accumulation. AB - Triclosan is an antimicrobial agent added to a wide array of consumer goods and personal care products. Through its use, it is introduced into municipal sewer systems where it is only partially removed during wastewater treatment. In this study, triclosan was measured in dated sediment cores from four urbanized estuaries in order to reconstruct temporal and spatial trends of accumulation. Measurable concentrations of triclosan first appeared in each of the sediment cores near 1964, which corresponds with the US patent issuance date of triclosan. The presence of triclosan at each of the study sites at or near the patent date indicates that long-term preservation is occurring in estuarine sediments. Temporal trends of triclosan at each location are unique, reflecting between site variability. Concentrations at one site climbed to as high as 400ngg(-1), due in part, to local commercial production of triclosan. At two locations, levels of triclosan rise towards the surface of each core, suggesting increasing usage in recent years. One location adjacent to a major combined sewer overflow had high sediment concentrations of triclosan, confirming their potential as a source of triclosan to estuaries. PMID- 20006372 TI - The influence of colloids on the geochemical behavior of metals in polluted water using as an example Yongdingxin River, Tianjin, China. AB - The role of colloids in estuarine and marine systems has been studied extensively in recent years, whereas less is known about the polluted freshwater system. Yongdingxin River is one of the major recipients of industrial effluents in Tianjin. This article evaluates the role of colloids in controlling geochemical behavior of Cu, Zn, Fe, Mn, Hg and Cr at the confluences between Yongdingxin River and its major tributaries Beijing Drainage River, Jinzhong River and Beitang Drainage River. Based on the distribution of metal partitioning among particulate (>0.22mum), colloidal (1kDa to 0.22mum) and truly dissolved (<1kDa) fractions, the metals can be assigned to the following groups: Group 1 - organic colloidal pool-borne elements Cu and Cr; Group 2 - inorganic colloidal pool-borne metals Fe and Mn; Group 3 - Zn and Hg characterized by varying complexation patterns. The distribution of metal partitioning among particulate, colloidal and truly dissolved fractions was influenced by anthropogenic input. In addition, the theoretical concentrations of elements in case of conservative mixing between the waters of Yongdingxin River and the waters of its tributaries were compared with the measured values to evaluate the geochemical role of colloids. The result showed that all of the metals presented a non-conservative mixing behavior. Addition of colloids resulted in the removal of metals from the water column to bed sediment during river water mixing, which was furthermore confirmed by the similar partition coefficient of metal concentration between colloid and sediment. PMID- 20006373 TI - Improving spatial concentration estimates for nitrogen oxides using a hybrid meteorological dispersion/land use regression model in Los Angeles, CA and Seattle, WA. AB - Predictions from a simple line source dispersion model, Caline3, were included as a covariate in a land use regression (LUR) model for NO(X)/NO(2) in Los Angeles, CA and Seattle, WA. The Caline3 model prediction assumed a unit emission factor for all roadway segments (1.0g/vehicle-mile). The NO(X) and/or NO(2) measurements for LA and Seattle were obtained from a comprehensive measurement campaign that is part of the Multi-Ethnic Study of Atherosclerosis Air Pollution Study (MESA Air). The measurement campaigns in both cities were approximately 2weeks in duration employing approximately 145 measurement sites in Greater LA and 26 sites in Seattle. The best "standard" LUR model (obtained without the inclusion of the Caline3 predictions) in LA had R(2) values of 0.53 for NO(X) and 0.74 for NO(2). The leave-one-out cross-validated R(2) values for NO(X) and NO(2) were 0.45 and 0.71, respectively. The equivalent "standard" NO(2) model for Seattle had an R(2) of 0.72 and a leave-one-out cross-validated R(2) of 0.63. When the Caline3 variable was included in the LA hybrid model, the R(2) values were 0.71 and 0.79 for NO(X) and NO(2), respectively. The corresponding cross-validated R(2) values were 0.66 and 0.77, for NOX and NO2, respectively. In Seattle, the inclusion of the Caline3 variable resulted in a NO(2) model with an R(2) of 0.81 and a corresponding cross-validated R(2) of 0.67. In LA, hybrid model performance was not affected by excluding roadways with annual average daily traffic volumes (AADT)<100,000. When the Caline3 predictions for heavy-duty trucks and lighter duty vehicles were modelled as separate terms, the estimated fleet average NO(X) emission factors were 8.9 (SE=0.7) and 0.16 (SE=0.12) grams NO(X)/vehicle mile for heavy-duty and lighter-duty vehicles, respectively. These values are consistent with fleet average emission factors computed for LA with EMFAC 2007. PMID- 20006374 TI - Toxicity of three phenolic compounds and their mixtures on the gram-positive bacteria Bacillus subtilis in the aquatic environment. AB - Although phenolic compounds are intensively studied for their toxic effects on the environment, the toxicity of catechol, resorcinol and hydroquinone mixtures are still not well understood because most previous bioassays are conducted solely using single compound based on acute tests. In this work, the adverse effect of individual phenolic compounds (catechol, resorcinol and hydroquinone) and the interactive effect of the binary and tertiary mixtures on Bacillus subtilis (B. subtilis) using microcalorimetric method were examined. The toxicity of individual phenolic compounds follows the order catechol>resorcinol>hydroquinone with their respective half inhibitory concentration as 437, 728 and 934 microg mL(-)(1). The power-time curve of B. subtilis growth obtained by microcalorimetry is in complete agreement with the change in turbidity of B. subtilis against time, demonstrating that microcalorimetric method agrees well with the routine microbiological method. The toxicity data obtained from phenolic compound mixtures show that catechol and hydroquinone mixture possess synergistic effect while the other mixtures display additive joint actions. Furthermore, the concentration addition (CA) and independent action (IA) models were employed to predict the toxicities of the phenolic compounds. The experimental results of microcalorimetry show no significant difference on the toxicity of the phenolic compound mixtures from that predicted by CA. However, IA prediction underestimated the mixture effects in all the experiments. PMID- 20006375 TI - Trends of legacy and new persistent organic pollutants in the circumpolar arctic: overview, conclusions, and recommendations. AB - This article provides an overview of key findings in the reviews in this special issue on the assessment of persistent organic pollutants (POPs) under the Arctic Monitoring and Assessment Program (AMAP), identifies knowledge gaps, and presents conclusions and recommendations for future work. The articles in this special issue summarize the peer reviewed literature and selected technical reports on trends of concentrations and possible biological effects of POPs in the Arctic published up to early 2009. PMID- 20006376 TI - p53 alteration independently predicts poor outcomes in patients with endometrial cancer: a clinicopathologic study of 131 cases and literature review. AB - OBJECTIVE: The aim of this study was to evaluate the prognostic impact of p53 alteration in human uterine endometrial adenocarcinoma. METHODS: One hundred and thirty-one patients with primary endometrial adenocarcinoma were included in the study. The p53 mutation and/or protein expression were evaluated by polymerase chain reaction-single-strand conformational polymorphism and by immunohistochemical analysis, respectively. Clinical and pathological parameters were obtained from medical records. Survival data were estimated using Kaplan Meier estimates and compared with the log-rank test where indicated. Multivariate analysis was performed using the Cox regression method. RESULTS: Thirty nine cases (29.8%) containing p53 alterations had a lower disease specific-survival rate and disease-free survival rate than those without p53 alterations. Statistically significant correlations were seen between p53 alteration and non endometrioid histology type, high grade tumors, and the absence of progesterone receptor. Multivariate analyses showed that both p53 alteration and FIGO stage at diagnosis were adverse prognostic factors. The group of women with p53 alteration had an 11.0-fold increased risk of disease specific death (95% confidence interval: 1.008-120.765) compared to women whose tumors lacked p53 alteration. CONCLUSION: p53 alteration defines a subset of endometrial adenocarcinoma with highly aggressive behavior and predicts lower survival in patients with endometrial adenocarcinoma. PMID- 20006377 TI - Culling intervals and culling risks in four stages of the reproductive life of first service and reserviced female pigs in commercial herds. AB - The objectives of this study were to measure culling intervals and culling risks in the four stages of the reproductive life of female pigs and to compare culling intervals between the number of services and between herd groups, based on herd productivity. We also compared survival patterns of females pigs between these herd groups. Our data set included lifetime records of 52,792 females born between 2001 and 2004 in 101 commercial herds. Two herd groups were selected on the basis of the upper 25th percentile of pigs weaned per mated female per 5 yr between 2002 and 2006, namely the high-performing herds, and ordinary herds. Culled females were also allocated into four groups based on the stages of their reproductive life when culled: unmated gilts, mated gilts, unmated sows, and mated sows. Culling intervals in unmated gilts and mated gilts were defined as the number of days from birth to culling and from first mating to culling, respectively. Culling intervals in unmated sows and mated sows were the number of days from weaning to culling. The number of services was categorized into two groups: first service and reservice groups. Multilevel linear mixed-effects models and survival analysis were performed. Culling intervals (+/-SEM) in unmated gilts, mated gilts, unmated sows, and mated sows were 302.9+/-1.16, 98.4+/-0.92, 14.3+/-0.12, and 89.6+/-0.42 d, respectively. Culling risks in the four groups were 5.6%, 7.1%, 58.0%, and 29.3%, respectively. In unmated gilts, mated gilts, and mated sows, the culling intervals in the high-performing herds were 43.0, 18.9, and 16.0 d shorter than those in ordinary herds, respectively (P<0.05), but no difference was found between the herd groups for the culling interval of unmated sows. For mated sows in the reservice group, culling intervals of high-performing herds were >or=13.7 d shorter than those of the ordinary herds (P<0.05), but for mated sows in the first service group, there was no difference in the culling interval between the herd groups. The culling hazard from 8 wk postweaning for mated sows in high-performing herds increased more rapidly than that in ordinary herds. In conclusion, to reduce culling intervals and improve herd productivity, we recommend implementing a strict culling policy for mated gilts and mated sows, especially reserviced females. PMID- 20006378 TI - Sentence integration processes: an ERP study of Chinese sentence comprehension with relative clauses. AB - In an event-related potentials (ERPs) study, we examined the comprehension of different types of Chinese (Mandarin) relative clauses (object vs. subject extracted) to test the universality and language specificity of sentence comprehension processes. Because Chinese lacks morphosyntactic cues to sentence constituent relations, it allows a test of the possibility that semantic contextual processes dominate the extraction of clausal relations, in contrast to the structure-dependent processing in English and many other languages. ERP results at the RC embedded verbs showed a P600 effect for the subject-extraction type, reflecting a processing of phrasal reconfiguration, and an N400 effect for the object-extraction type, reflecting a processing of meaning reinterpretation. A central-frontal sustained negativity was produced by the RC head noun of object extraction, suggesting a combined effect of meaning derivation and referents establishment. LORETA (Low Resolution Electrical Tomography) source localization showed activation of posterior dominance (e.g., BA 22/39/19/41/42) supporting the integration of structure mapping (P600) and meaning derivation (N400) in a developing sentential representation, consistent with the memory unification and control model (Hagoort, 2005). More left-lateralized anterior regions of a frontal-temporal network (e.g., BA 47/38) became active later in the sentence (a sustained central-frontal negativity), when the thematic-role specification for multiple referents may have required additional cognitive and memory resources. Our findings suggest that Chinese sentence reading recruits a neural network that is sensitive to the sequential/hierarchical organization of linguistic inputs in a manner that resembles to the structure-dependent cognitive processes in other languages, reflecting a universal property of language processing. The ERP data shows that early lexical processes are important in the integration process, but also challenges the view that Chinese text reading depends primarily on semantic contextual processing in the derivation of meaning representation. PMID- 20006379 TI - Relationship between bond-strength tests and clinical outcomes. AB - One often alleges that laboratory bond-strength testing cannot predict clinical effectiveness of adhesives. Major argument to sustain this claim is the wide variation in bond-strength values recorded for one specific adhesive among different research institutes worldwide. The main reason for these inconsistent bond-strength measurements is supposedly the current lack of a standard bond strength testing protocol. This paper (and presentation) aimed to report on an extensive literature review with regard to the different laboratory bond-strength test methods and their data provided, along with a second extensive literature review on clinical effectiveness data of adhesives in terms of retention rates of adhesive Class-V restorations. Combining both systematic reviews, we have subsequently searched for a potential relationship between bond-strength data and clinical outcomes. PMID- 20006380 TI - Selective cell affinity of biomimetic micro-nano-hybrid structured TiO2 overcomes the biological dilemma of osteoblasts. AB - OBJECTIVE: There is a great demand for dental implant surfaces to accelerate the process of peri-implant bone generation to reduce its healing time and enable early loading. To this end, an inverse correlation between the proliferation and functional maturation (differentiation) in osteoblasts presents a challenge for the rapid generation of greater amounts of bone. For instance, osteoblasts exhibit faster differentiation but slower proliferation on micro-roughened titanium surfaces. Using a unique micro-nano-hierarchical topography of TiO(2) that mimics biomineralized matrices, this study demonstrates that this challenge can be overcome without the use of biological agents. METHODS: Titanium disks of grade 2 commercially pure titanium were prepared by machining (smooth surface). To create a microtexture with peaks and valleys (micropit surface), titanium disks were acid-etched. To create 200-nm TiO(2) nanonodules within the micropits (nanonodule-in-micropit surface), TiO(2) was sputter-deposited onto the acid etched surface. Rat bone marrow-derived osteoblasts and NIH3T3 fibroblasts were cultured on machined smooth, micropit, and nanonodule-in-micropit surfaces. RESULTS: Despite the substantially increased surface roughness, the addition of 200-nm nanonodules to micropits increased osteoblast proliferation while enhancing their functional differentiation. In contrast, this nanonodule-in micropit surface decreased proliferation and function in fibroblasts. SIGNIFICANCE: The data suggest the establishment of cell-selectively functionalized nano-in-micro smart titanium surfaces that involve a regulatory effect on osteoblast proliferation, abrogating the inhibitory mechanism on the micropitted surface, while enhancing their functional differentiation. Biomimetic and controllable nature of this nanonodules-in-micropits surface may offer a novel micro-to-nanoscale hierarchical platform to biologically optimize nanofeatures of biomaterials. Particularly, this micro-nano-hybrid surface may be an effective approach to improve current dental implant surfaces for accelerated bone integration. PMID- 20006381 TI - The promotion of endothelial progenitor cells recruitment by nerve growth factors in tissue-engineered blood vessels. AB - Endothelial progenitor cells (EPCs) mobilization and homing are critical to the development of an anti-thrombosis and anti-stenosis tissue-engineered blood vessel. The growth and activation of blood vessels are supported by nerves. We investigated whether nerve growth factors (NGF) can promote EPCs mobilization and endothelialization of tissue-engineered blood vessels. In vitro, NGF promoted EPCs to form more colonies, stimulated human EPCs to differentiate into endothelial cells, and significantly enhanced EPCs migration. Flow cytometric analysis revealed that NGF treatment increased the number of EPCs in the peripheral circulation of C57BL/6 mice. Furthermore, the treatment of human EPCs with NGF facilitated their homing into wire-injured carotid arteries after injection into mice. Decellularized rat blood vessel matrix was incubated with EDC cross-linked collagen and bound to NGF protein using the bifunctional coupling agent N-succinmidyl3-(2-pyridyldit-hio) propionate (SPDP). The NGF-bound tissue-engineered blood vessel was implanted into rat carotid artery for 1 week and 1 month. NGF-bound blood vessels possessed significantly higher levels of endothelialization and patency than controls did. These results demonstrated that NGF can markedly increase EPCs mobilization and homing to vascular grafts. Neurotrophic factors such as NGF have a therapeutic potential for the construction of tissue-engineered blood vessels in vivo. PMID- 20006382 TI - Synthesizing and binding dual-mode poly (lactic-co-glycolic acid) (PLGA) nanobubbles for cancer targeting and imaging. AB - Accurate assessment of tumor boundaries and recognition of occult disease are important oncologic principles in cancer surgeries. However, existing imaging modalities are not optimized for intraoperative cancer imaging applications. We developed a nanobubble (NB) contrast agent for cancer targeting and dual-mode imaging using optical and ultrasound (US) modalities. The contrast agent was fabricated by encapsulating the Texas Red dye in poly (lactic-co-glycolic acid) (PLGA) NBs and conjugating NBs with cancer-targeting ligands. Both one-step and three-step cancer-targeting strategies were tested on the LS174T human colon cancer cell line. For the one-step process, NBs were conjugated with the humanized HuCC49 Delta C(H)2 antibody to target the over-expressed TAG-72 antigen. For the three-step process, cancer cells were targeted by successive application of the biotinylated HuCC49 Delta C(H)2 antibody, streptavidin, and the biotinylated NBs. Both one-step and three-step processes successfully targeted the cancer cells with high binding affinity. NB-assisted dual-mode imaging was demonstrated on a gelatin phantom that embedded multiple tumor simulators at different NB concentrations. Simultaneous fluorescence and US images were acquired for these tumor simulators and linear correlations were observed between the fluorescence/US intensities and the NB concentrations. Our research demonstrated the technical feasibility of using the dual-mode NB contrast agent for cancer targeting and simultaneous fluorescence/US imaging. PMID- 20006383 TI - Structural analysis of three peptides related to the transmambranic helix VI of AT1 receptor. AB - INTRODUCTION: Angiotensin II (AII) is the main active product of the renin angiotensin system. Better known effects of AII are via AT1 receptor (AT1R). Expression of AT1R mutants (L265D and L262D) in CHO cells increased cAMP formation when compared to CHO cells expressing the wild type (WT) AT1R. Morphological transformation of CHO cells transfected with mutants correlated with their increased cAMP formation. DNA synthesis was inhibited in these cells too, indicating that cAMP promotes inhibitory effects on transfected CHO cells growth and causes their morphological change from a tumorigenic phenotype to a non-tumorigenic one. OBJECTIVES: To assess the importance of leucine 262 and 265 in determining AT1R structure by means of a comparative structural analysis of two mutant peptides and of a wild-type fragment. METHODOLOGY: Three peptides had their conformation compared by circular dichroism (CD): L262D(259-272), L265D(259 272) (mutants) and WT(260-277). RESULTS: Secondary structures were: beta-turn for WT and L262D and random coil for L265D. CONCLUSIONS: Strong correlation was found in the results of biochemical, cellular and structural approaches used to compare WT AT1R to mutant types. Random coil structure of the L265D mutant may be a key point to explain those changes observed in biochemical (binding and signal transduction) and proliferation assays (Correa et al., 2005). beta-Turn formation is an important step during early protein folding and this secondary simple structure is present in L262D and WT, but not in L265D. Therefore, leucine 265 seems to play a crucial role in determining an entirely functional AT1R. PMID- 20006384 TI - Improved 3D-QSAR analyzes for the predictive toxicology of polybrominated diphenyl ethers with CoMFA/CoMSIA and DFT. AB - With the popular methods of CoMFA and CoMSIA, three-dimensional quantitative structure-activity relationships (QSARs) were newly developed for the toxicity of polybrominated diphenyl ethers (PBDEs). The choice of optimized geometries by density functional theory (DFT) as molecular template and the RMSD-based molecular alignment strategy might mostly contribute to the QSAR improvement, which was highlighted specifically by the increased q2 of 0.870 for CoMFA, 0.887 for CoMSIA, respectively. QSARs analyzes indicated that the steric effects from ortho- and meta-substitution and the correlated hydrophobicities have the greatest impact on the binding affinities of aryl hydrocarbon receptor (AhR) to PBDEs. Though the effects of electrostatics were comparatively inferior in the AhR binding, the aromatic interaction and possible charge transfer proved to be indispensable for toxicity mediation. Consistent with that proposed previously for other structurally similar compounds, such as dioxins and polychlorinated biphenyls, the predictive toxicology was helpful to understand the congener specificity of toxicity of PBDEs. PMID- 20006385 TI - Construction of single-chain Fv with two possible CDR3H conformations but similar inter-molecular forces that neutralize bovine herpesvirus 1. AB - Bovine herpesvirus 1 (BoHV-1) causes respiratory and genital diseases in cattle for which available vaccines do not confer adequate protection. Since passive immunization with antibodies permits disease prevention, single-chain fragment variable (scFv), originating from a monoclonal bovine IgG1 antibody against BoHV 1, were constructed and expressed in Pichia pastoris in V(lambda)-V(H) orientation via a flexible seven-amino acid linker. Similar to the intact IgG, the purified recombinant scFv neutralized BoHV-1 in vitro and recognized viral antigens in BoHV-1 infected MDBK cells by immunofluorescence. Homology modeling of the Fv predicts two distinct conformations for CDR3H. Firstly, a long protruding CDR3H conformation where no disulfide linkage occurred between two "non-canonical" Cys residues resulted in a large binding cavity between V(lambda) and V(H). Secondly, a smaller potential antigen-binding cavity is predicted with a disulfide linkage between the two Cys residues of CDR3H creating a six-membered loop in the ascending polypeptide, which fitted into the space between V(lambda) and V(H). Despite such potential configurational diversity of the antigen-binding site, the electrostatic surface potentials that would interact with the BoHV-1 epitope are largely similar for both the topographies where salt-bridge type electrostatic interactions likely occur at the edges of the binding site. Given that IgG1 antibody against BoHV-1 is clonally selected, it is likely that disulfide-stabilized broader and flatter surface topography is specifically generated to accommodate the predicted carbohydrate neutralizing B-epitope on BoHV-1. The specificity and neutralizing capacity for BoHV-1 of the scFv should make this bovine antibody fragment a useful diagnostic and potential therapeutic candidate for an important viral pathogen in cattle. PMID- 20006386 TI - The CES-D in Chinese American women: construct validity, diagnostic validity for major depression, and cultural response bias. AB - Previous studies of the Center for Epidemiologic Studies Depression Scale (CES-D) in Chinese Americans describe internal reliability and factor structure. We report CES-D construct validity and diagnostic validity for major depression in a probability sample of 168 community-dwelling Chinese American women. Internal consistency was satisfactory (Cronbach's alpha=0.86). Good construct validity was indicated by significantly higher mean CES-D scores for respondents who reported lower social support, worse self-perceived general health, or stressful life events, including intimate partner violence. Cultural response bias was found, with positively-stated CES-D items (e.g. "I was happy") producing higher depression scores in immigrants and subjects who preferred to speak Chinese. Diagnostic validity for major depression was assessed using the Composite International Diagnostic Interview. A CES-D cut-off score of 16 had sensitivity of 100% (95% CI: 44% to 100%), specificity of 76% (95% CI: 69% to 82%), PPV of 7% (95% CI: 3% to 19%) and NPV of 100% (95% CI: 97% to 100%). Our findings suggest that the CES-D is useful for screening out non-depressed subjects in a first stage assessment. However, it should be followed by a diagnostic tool in Chinese American women with scores above the cut-off in order to identify those with clinical depression. PMID- 20006387 TI - HLA-DRB1 allele heterogeneity influences multiple sclerosis severity as well as risk in Western Australia. AB - Susceptibility to multiple sclerosis (MS) has been consistently associated with the Human Leukocyte Antigen (HLA)-DRB11501 genotype, however effects on disease severity and clinical outcome have varied in different populations. We present the results of a high-resolution HLA-DRB1 genotyping and genotype-phenotype correlation study in a large West Australian MS cohort. Our findings indicate that in this population, which is of largely Anglo-Celtic and Northern European origin, HLA-DRB11501 is not only a strong determinant of disease risk but may also be associated with disease severity as measured by the Multiple Sclerosis Severity Score (MSSS), with the MSSS increasing by an estimated 0.51 per DRB11501 allele. We also found evidence that the HLA-DRB11201 allele is associated with less severe disease. PMID- 20006388 TI - Anti-ganglioside complex IgM antibodies in multifocal motor neuropathy and chronic immune-mediated neuropathies. AB - Anti-ganglioside complexes (GSCs) IgG antibodies have been reported in patients with Guillain-Barre (GBS) or Fisher syndrome but little is known on their presence in multifocal motor neuropathy (MMN) or other chronic immune-mediated neuropathies. We examined 24 patients with MMN, 34 with chronic inflammatory demyelinating polyradiculoneuropathy (CIDP), 23 with neuropathy associated with IgM monoclonal gammopathy (PN+IgM), 13 with GBS, 34 with motor neuron disease (MND), 24 with other neuropathies and 20 normal subjects. Patients' sera were tested by ELISA for IgM reactivity to GM1, GM2, GD1a, GD1b and GT1b and with GSCs made by any combination of two of these gangliosides. In all GM1 positive patients with MMN (11), PN+IgM (1), CIDP (1) and POEMS (1), binding to GM1 was abolished or consistently reduced when tested in GSCs also containing GD1a or other gangliosides. This only occurred in one of the three GM1 positive MND patients. In a patient with PN-IgM and anti-GM2 and GD1a IgM, both reactivities were reduced when tested in GSCs also containing GM1. New reactivities were found in a patient with CIDP and anti-GD1b IgM who presented an additional reactivity to GT1b/GM1 and GT1b/GM2 GSCs, and in one with PN-IgM who had reactivity to GM2/GD1b but not to individual gangliosides. Testing for IgM antibodies to GSCs rarely permitted to identify new reactivities in chronic immune neuropathies. IgM binding to gangliosides was however often modified in GSCs suggesting that these reactivities may be affected by contiguous gangliosides possibly influencing their pathogenicity. PMID- 20006389 TI - Bilateral versus unilateral cochlear implantation in young children. AB - OBJECTIVES: To compare the preverbal communication skills of two groups of young implanted children: those with unilateral implantation and those with bilateral implantation. MATERIAL AND METHODS: The study assessed 69 children: 42 unilaterally and 27 bilaterally implanted with age at implantation less than 3 years. The preverbal skills of these children were measured before and 1 year after implantation, using Tait Video Analysis that has been found able to predict later speech outcomes in young implanted children. RESULTS: Before implantation there was no significant difference between the unilateral group and the bilateral group. There was still no difference at 12 months following implantation where vocal autonomy is concerned, but a strongly significant difference between the groups for vocal turn-taking and non-looking vocal turns, the bilateral group outperforming the unilateral group. Regarding gestural turn taking and gestural autonomy, there was a strongly significant difference between the two groups at the 12 month interval, and also a difference before implantation for gestural autonomy, the unilateral group having the higher scores. Multiple regression of non-looking vocal turns revealed that 1 year following implantation, bilateral implantation contributed to 51% of the variance (p<0.0001), after controlling for the influence of age at implantation and length of deafness which did not reach statistical significance. CONCLUSIONS: Profoundly deaf bilaterally implanted children are significantly more likely to use vocalisation to communicate, and to use audition when interacting vocally with an adult, compared with unilaterally implanted children. These results are independent of age at implantation and length of deafness. PMID- 20006390 TI - The effects of 17beta-estradiol on various reproductive parameters in the hermaphrodite fish Kryptolebias marmoratus. AB - The effect of a single injection of 17beta-estradiol (E2) was evaluated in the hermaphrodite fish Kryptolebias marmoratus. The fish [average body weight (BW), 0.15+/-0.01 g] were injected with either two concentrations of E2 (1 and 100 microg/g BW) once intraperitoneally. They were sampled at intervals of 7, 15, and 30 days after a single E2 injection. Gonadosomatic index (GSI), hepatosomatic index (HSI), the frequency of gonadal development, number of ovulated eggs, and plasma steroids levels were measured. The transcript abundances of vitellogenin (VTG) and estrogen receptors (ERalpha and beta) mRNA in the liver were also analyzed using quantitative real time polymerase chain reaction (real time PCR). GSI and the frequency of mature oocytes in the 100-microg E2-exposed group decreased compared to that of the control group during the experiment, and the number of ovulated eggs in the 100-microg E2-exposed group was lower when compared to the other groups. However, plasma E2 and 11-ketotestosterone (11-KT) levels were not significantly different between the experimental groups. On the other hand, plasma testosterone level and VTG mRNA abundance in the 100-microg E2 exposed group were significantly lower than the control group after 30 days. These results indicate that E2 stimulation at high concentration interferes with reproductive phenomena through delayed response. In addition, HSI in the 100 microg E2-exposed group and ERalpha mRNA abundance in the 1-microg E2-exposed group were significantly higher than the control group at 7 days after E2 injection, although there was no significant difference in HSI and ERalpha mRNA between all groups at 30 days. These results indicate temporal responses in reproductive parameters following high-dose E2 exposure in the hermaphrodite fish K. marmoratus. PMID- 20006391 TI - Medical healthcare utilization as related to long-term care at home or in special accommodation. AB - This study aimed to investigate medical healthcare utilization 3-5 years following the decision about long-term care at home vs. in special accommodation in older people. A total of 1079 people who were granted long-term care the years 2001, 2002 or 2003 were studied regarding the number of hospital stays and the number of contacts with physicians in outpatient care in the 3-5 subsequent years. Those living at home and those in special accommodation were compared regarding medical healthcare utilization during the 3-5 subsequent years. Data were collected through the study Good Aging in Skane (GAS) and through the registers, Patient Administrative Support in Skane (PASiS) and PrivaStat. Utilization of medical healthcare decreased slightly in the years following the decision about long-term care. Despite younger age and less dependency in activities of daily living (ADL), those living at home utilized hospital and outpatient care to a greater extent than those in special accommodation; these differences remained over time. Thus, it seems as long-term care needs to become more effective in the prevention of medical healthcare utilization among those cared for at home. More, older people who are granted long-term care at home may otherwise imply increased utilization of medical healthcare. PMID- 20006392 TI - Prognostic impact of peritonealisation in rectal cancer treated with preoperative chemoradiotherapy: extraperitoneal versus intraperitoneal rectal cancer. AB - BACKGROUND AND PURPOSE: The oncologic outcomes of extraperitoneal (EP) rectal cancer are known to differ from those of intraperitoneal (IP) rectal cancer; however, these differences have not been studied in rectal patients treated by preoperative chemoradiotherapy (CRT). The aim of this study is to evaluate the prognostic impact of peritonealisation in rectal patients treated by preoperative CRT. MATERIALS AND METHODS: This study analyzed the data of 362 patients who received preoperative CRT and underwent curative surgery for locally advanced rectal cancer at 3-9 cm above the anal verge. Patients were categorised into EP and IP groups based on whether peritonealisation was present, according to pathology reports. The oncologic outcomes between the two groups were compared. RESULTS: Peritonealisation was absent in 330 patients and present in 32 patients. In univariate analysis, disease-free survival was significantly worse in the EP group than in the IP group (73.0% versus 93.5%, p=0.035). Multivariate analysis revealed the following independent risk factors for recurrence: the absence of peritonealisation (p=0.023), ypT stage (p=0.015) and ypN stage (p<.0001). CONCLUSIONS: Peritonealisation of rectal cancer may be a prognostic factor of disease-free survival in patients with rectal cancer treated by preoperative CRT and surgery. PMID- 20006393 TI - Plasminogen activator inhibitor-1 (PAI-1) activity and retinal vascular calibre in type 2 diabetes. AB - OBJECTIVE: To describe relationships of retinal vascular calibre with plasminogen activator inhibitor-1 (PAI-1) and other cardiovascular risk factors in people with type 2 diabetes. METHODS: We recruited 112 community-based persons aged 44 83years with type 2 diabetes, photo-documented retinal status using a digital fundus camera, and measured traditional and novel vascular risk factors. Retinal arteriolar and venular calibre and the arterio-venous ratio (AVR) were determined from fundus photographs using a validated computer-assisted method. RESULTS: In adjusted linear regression models, PAI-1 activity was strongly associated with all measures of retinal vascular calibre: positively with arterioles (p=0.005) and AVR (p=0.001), and inversely with venules (p=0.001). In addition, wider arterioles were independently associated with waist-hip ratio (p<0.0001), HDL-C (p=0.015), and lower systolic blood pressure (p=0.042), whereas narrower venules were associated with older age and a higher albumin excretion rate. Neither arteriolar nor venular calibre was associated with plasma total homocysteine or C reactive protein concentration. CONCLUSION: Retinal vascular calibre is independently associated with PAI-1 activity in type 2 diabetes. This finding supports a role for PAI-1 activity in the microvasculature of persons with type 2 diabetes and may explain the link between retinal vascular calibre and cardiovascular disease. PMID- 20006394 TI - Efficacy of entecavir in treatment-naive patients with hepatitis B virus-related decompensated cirrhosis. AB - BACKGROUND & AIMS: The effect of entecavir (ETV) therapy on viral suppression and hepatic function in hepatitis B virus (HBV) patients with decompensated cirrhosis has not been established. We evaluated ETV as first-line monotherapy in these patients. METHODS: We consecutively enrolled 70 HBV-infected patients with decompensated cirrhosis primarily treated with 0.5mg/day ETV, and evaluated the clinical outcomes by intention-to-treat analyses. We also compared the virological responses of 55 patients treated for 12 months (decompensated group) with those of 144 chronic hepatitis or compensated cirrhosis patients (compensated group). RESULTS: The cumulative transplantation-free survival was 87.1% at 1year. ETV treatment for 12 months resulted in improved Child-Turcotte Pugh (CTP) and model for end-stage liver disease (MELD) scores. Sixty-six percent (36/55) of patients achieved CTP class A and 49% (27/55) showed improvement in the CTP score of 2 points after 12 months of ETV. The 1-year cumulative rates of HBV DNA negativity and HBeAg loss were 92.3% and 54.0%, respectively, by intention-to-treat analysis. The rates of HBV DNA negativity, HBeAg seroconversion/loss and ALT normalization at month 12 were similar for the decompensated and compensated groups. Cox regression analysis showed that pretreatment HBeAg seropositivity was a negative predictor of HBV DNA clearance during ETV therapy (hazard ratio, 0.514; 95% confidence interval 0.367-0.719; p<0.001). CONCLUSIONS: One-year initial ETV therapy was similarly effective in both compensated and decompensated liver disease HBV patients. In addition, it improved underlying liver function in decompensated patients. PMID- 20006395 TI - Focus. PMID- 20006396 TI - Effects of antibiotics on expression and function of Toll-like receptors 2 and 4 on mononuclear cells in patients with advanced cirrhosis. AB - BACKGROUND & AIMS: Toll-like receptors (TLRs) are critical to innate immune responses. TLR4 recognises Gram-negative bacteria, whilst TLR2 recognises Gram positive. We examined TLR expression and function in cirrhosis, and whether this is affected by antibiotic therapy. METHODS: Sixty-four subjects were included (23 controls and 41 Child-Pugh C cirrhotic patients). Thirty patients were taking norfloxacin or trimethoprim-sulfamethoxazole as prophylaxis against bacterial peritonitis and 11 were not. In a second study, 8 patients were examined before and after commencement of antibiotics. Monocyte expression of TLR2 and 4 was determined by flow cytometry. Monocytes from the patients with paired samples were stimulated using TLR ligands and TNF-alpha production measured. RESULTS: Patients not taking antibiotics had significantly decreased TLR4 expression compared with controls (0.74 vs. 1.0, p=0.009) and patients receiving antibiotics (0.74 vs. 0.98, p=0.02). There were no differences with regard to TLR2. In the patients with paired samples, TLR4 expression increased (0.74-1.49, p=0.002) following antibiotic use, whilst again, there was no change in TLR2 expression (0.99 vs. 0.92, p=0.20). TLR4-dependent TNF-alpha production increased following antibiotic use (1077 vs. 3620pg/mL, p<0.05), whilst TLR2-dependent production was unchanged. CONCLUSIONS: TLR4 expression is decreased in patients with Child-Pugh C cirrhosis, but is restored by antibiotics targeting enteric Gram-negative bacteria. TLR4-dependent cytokine production also increases significantly following antibiotic therapy. This suggests that the high incidence of Gram negative infection in cirrhotic patients is in part due to down-regulation of the TLR4-dependant immune response and that the efficacy of antibiotic prophylaxis is contributed to by modulation of innate immunity. PMID- 20006397 TI - Prospective comparison of two algorithms combining non-invasive methods for staging liver fibrosis in chronic hepatitis C. AB - BACKGROUND & AIMS: Non-invasive assessment of liver fibrosis is a challenging area. Several methods have been proposed in patients with chronic hepatitis C (CHC) but their performance may be improved when they are combined as suggested by recently proposed algorithms using either transient elastography (TE) and Fibrotest (FT) (Castera) or AST-to-Platelet Ratio Index (APRI) and FT (SAFE biopsy). The aim of this prospective study was to compare the performance of these two algorithms for diagnosing significant fibrosis and cirrhosis in 302 CHC patients. METHODS: All patients underwent TE, FT and APRI the same day as liver biopsy, taken as reference standard. RESULTS: Significant fibrosis (Metavir F>or=2) was present in 76% of patients and cirrhosis (F4) in 25%. TE failure was observed in eight cases (2.6%). For significant fibrosis, Castera algorithm saved 23% more liver biopsies (71.9% vs. 48.3%, respectively; p<0.0001) than SAFE biopsy but its accuracy was significantly lower (87.7% vs. 97.0%, respectively; p<0.0001). Regarding cirrhosis, accuracy of Castera algorithm was significantly higher than that of SAFE biopsy (95.7% vs. 88.7%, respectively; p<0.0001). The number of saved liver biopsies did not differ between the two algorithms (78.8% vs. 74.8%; p=NS). CONCLUSIONS: Both algorithms are effective for non-invasive staging of liver fibrosis in chronic hepatitis C. Although the number of liver biopsies avoided does not differ between algorithms for diagnosing cirrhosis, it is significantly higher with Castera algorithm than SAFE biopsy for significant fibrosis. PMID- 20006398 TI - Distinct antiviral signaling pathways in primary human hepatocytes and their differential disruption by HCV NS3 protease. AB - BACKGROUND & AIMS: Molecular sensors recognize viral nucleic acids and initiate events that subsequently enable cells to control and clear infection. Hepatitis C Virus (HCV) can interfere with the innate host response and the NS3/4A protease was reported to specifically block antiviral signaling pathways, a finding that had yet to be studied in human primary hepatocytes. METHODS: Freshly isolated human primary hepatocytes, transduced with a lentiviral vector expressing HCV NS3/4A were stimulated with extracellular and intracellular double-stranded RNA (dsRNA) and the innate immune antiviral genes were quantified by quantitative PCR and microarrays analysis. RESULTS: We demonstrate that sensing receptors of human hepatocytes in primary cultures are stimulated following recognition of either mode of dsRNA delivery, inducing transcriptional up-regulation (over 100-fold) of multiple immune genes, either selectively or independently of recognition pathways. We also report that the intracellular dsRNA-activated innate response is severely compromised upon ectopic expression of the HCV NS3/4A protease gene in normal human primary hepatocytes, and completely restored by treatment with the NS3/4A protease specific inhibitor BILN2061. CONCLUSIONS: The present study indicates that NS3/4A has a wider protease-dependent effect on the intracellular Pathogen Recognition Receptor (PRR)-mediated immune response than on its extracellular counterpart, which underlies the major role of cytosolic dsRNA receptors in HCV recognition by primary human hepatocytes. PMID- 20006399 TI - Liver cancer: from molecular pathogenesis to new therapies: summary of the EASL single topic conference. PMID- 20006400 TI - Entecavir in decompensated HBV cirrhosis: the future is looking brighter. PMID- 20006401 TI - Will nano-fibers permit to turn liver cell transplantation into a curative tool against liver failure? PMID- 20006402 TI - EpCAM, a new marker for cancer stem cells in hepatocellular carcinoma. AB - BACKGROUND & AIMS: Cancer progression/metastases and embryonic development share many properties including cellular plasticity, dynamic cell motility, and integral interaction with the microenvironment. We hypothesized that the heterogeneous nature of hepatocellular carcinoma (HCC), in part, may be owing to the presence of hepatic cancer cells with stem/progenitor features. METHODS: Gene expression profiling and immunohistochemistry analyses were used to analyze 235 tumor specimens derived from 2 recently identified HCC subtypes (EpCAM(+) alpha fetoprotein [AFP(+)] HCC and EpCAM(-) AFP(-) HCC). These subtypes differed in their expression of AFP, a molecule produced in the developing embryo, and EpCAM, a cell surface hepatic stem cell marker. Fluorescence-activated cell sorting was used to isolate EpCAM(+) HCC cells, which were tested for hepatic stem/progenitor cell properties. RESULTS: Gene expression and pathway analyses revealed that the EpCAM(+) AFP(+) HCC subtype had features of hepatic stem/progenitor cells. Indeed, the fluorescence-activated cell sorting-isolated EpCAM(+) HCC cells displayed hepatic cancer stem cell-like traits including the abilities to self renew and differentiate. Moreover, these cells were capable of initiating highly invasive HCC in nonobese diabetic, severe combined immunodeficient mice. Activation of Wnt/beta-catenin signaling enriched the EpCAM(+) cell population, whereas RNA interference-based blockage of EpCAM, a Wnt/beta-catenin signaling target, attenuated the activities of these cells. CONCLUSIONS: Taken together, our results suggest that HCC growth and invasiveness is dictated by a subset of EpCAM(+) cells, opening a new avenue for HCC cancer cell eradication by targeting Wnt/beta-catenin signaling components such as EpCAM. PMID- 20006403 TI - Insulin resistance predicts rapid virological response in non-diabetic, non cirrhotic genotype 1 HCV patients treated with peginterferon alpha-2b plus ribavirin: some issues. PMID- 20006404 TI - Insulin resistance and HCV virologic response to peg-interferons (Peg-IFN) with ribavirin (RBV) in HIV/HCV co-infected patients. PMID- 20006405 TI - The future of coral reefs: a microbial perspective. AB - Microbial communities respond and quickly adapt to disturbance and have central roles in ecosystem function. Yet, the many roles of coral-associated microbial communities are not currently accounted for in predicting future responses of reef ecosystems. Here, we propose that a clearer understanding of coral associated microbial diversity and its interaction with both host and environment will identify important linkages occurring between the microbial communities and macroecological change. Characterizing these links is fundamental to understanding coral reef resilience and will improve our capacity to predict ecological change. PMID- 20006406 TI - Vitellogenin is an acute phase protein with bacterial-binding and inhibiting activities. AB - Previous studies have shown that vitellogenin (Vg) is an immune-relevant molecule, but its potential immunological role in vivo remains obscure. We demonstrated here that injection of lipopolysaccharide (LPS) and lipoteichoic acid (LTA) into male Danio rerio rapidly induced a significant up-regulation of Vg at both transcriptional and translational levels, and that serum Vg produced was able to bind to both Escherichia coli and Staphylococcus aureus and to inhibit their growth in a dose-dependent manner. All these data suggest that serum Vg in zebrafish D. rerio is an acute phase protein with bacterial-binding and inhibiting activities. It also bolsters the notion that factors normally involved in control of female reproduction are linked with immunity in organisms that rely on Vg for oocyte development. PMID- 20006408 TI - Outbreak of vancomycin-resistant enterococci in a tertiary hospital: the lack of effect of measures directed mainly by surveillance cultures and differences in response between Enterococcus faecium and Enterococcus faecalis. AB - To describe the effect of active surveillance to control vancomycin-resistant enterococci (VRE) after an outbreak, 549 surveillance rectal cultures were performed in 308 patients (35% positive). An educational intervention to prevent transmission was implemented. Infection and colonization by VR- Enterococcus faecalis decreased, but Enterococcus faecium persisted despite control measures. Infections by VR-E faecalis fell to zero in 2008. We observed difficulties in controlling colonization with measures directed mainly by surveillance cultures and differences between responses of E faecium and E faecalis. PMID- 20006407 TI - Expression of proliferative biomarkers in anal intraepithelial neoplasia of HIV positive men. AB - BACKGROUND: Anal intraepithelial neoplasia (AIN), a human papillomavirus (HPV) associated precursor lesion of anal carcinoma, is highly prevalent among HIV infected individuals, especially in men having sex with men (MSM). Early diagnosis and treatment of AIN might prevent development of anal cancer. OBJECTIVES: We aimed to evaluate the expression of 8 promising proliferative biomarkers in anal dysplasia and to compare the efficacy of these markers in diagnosing high-grade AIN. METHODS: Immunohistochemical analysis of minichromosome maintenance proteins (MCM3, MCM4, MCM6, and MCM7), p21, Ki-67, p16, and proliferating cell nuclear antigen (PCNA) was performed in a total of 49 specimens of normal anal mucosa and high- and low-grade anal dysplasia. HPV typing for 36 high- and low-risk HPVs was performed, and high-risk HPV-DNA loads were determined by real-time polymerase chain reaction (PCR) for HPV-types 16, 18, 31, and 33. RESULTS: A total of 392 immunohistochemical slides were analyzed in this study. In the progression from normal epithelium to high-grade dysplasia, we found significant differences in the expression of all biomarkers. A cutoff of 25% or 50% lesional immunopositivity for the 4 MCMs, Ki-67, and p16 resulted in 100% sensitivity and 100% specificity to diagnose high-grade AIN. Sensitivity and specificity of PCNA and p21 for a high-grade AIN diagnosis were lower. HPV-DNA was detectable in 100% of high-grade AIN and 87.5% of low-grade AIN lesions. All MCMs, p16, Ki-67, and PCNA, but not p21 correlated with cumulative lesional high grade HPV-DNA loads. LIMITATIONS: The relatively small number of samples is a limitation, especially for adequate subgroup analyses. CONCLUSIONS: MCMs, Ki67, and p16 are reliable immunohistochemical adjuncts for diagnosing high-grade AIN. PMID- 20006409 TI - Effectiveness of a catheter-associated bloodstream infection bundle in a Thai tertiary care center: a 3-year study. AB - BACKGROUND: We sought to determine the long-term impact of "bundled" infection control interventions on the rates of catheter-associated bloodstream infection (CA-BSI) in a middle-income country. SETTING: A 500-bed tertiary care center in Thailand. METHODS: A 3-year, hospital-wide, prospective quasi-experimental study was conducted for 1 year before the intervention (period 1), 1 year after implementation of the CA-BSI bundle (period 2), and at a 1-year follow-up after the intervention with intensified hand hygiene promotion (period 3). RESULTS: In period 1, 88 episodes of CA-BSI (14 cases per 1000 catheter-days) were recorded. During period 2, the CA-BSI rate decreased by 54.1 % (6.4 cases per 1000 catheter days; P <.001). Compared with period 1 (8% adherence), hand hygiene adherence was improved in period 2 (24%; P <.001) and period 3 (54%; P <.001). The CA-BSI rate was further decreased by 78% (1.4 cases per 1000 catheter-days; P <.001) during period 3. Notably, no CA-BSIs were seen in 6 of the 12 months (50%) of period 3. Compared with period 1, the mean number of catheter-days was significantly reduced in period 2 (4.9 +/- 1.5 days; P <.001) and period 3 (4.1 +/- 1.1 days; P <.001). CONCLUSION: Bundled infection control practices are feasible and effective in sustaining reduced incidence of CA-BSI in patients with central venous catheters in a resource-limited setting. PMID- 20006410 TI - Effect of surfactants, temperature, and sonication on the virucidal activity of polyhexamethylene biguanide against the bacteriophage MS2. AB - BACKGROUND: Virucidal compounds are essential in preventing the transmission of viral infection in the health care environment. Understanding their mechanisms of action is necessary to improve their efficacy. Inactivation of viruses is less documented than that of bacteria notably because different types of virus have diverse response to microbicides, making difficult to establish an inactivation pattern. METHODS: The effect of viral aggregates on the virucidal activity of polyhexamethylene biguanide-based microbicide VANTOCIL TG (Arch Chemicals, Manchester, UK) against the bacteriophage MS2 was investigated by using in combination a standard suspension efficacy test under different conditions and dynamic light scattering measuring the presence and size of aggregates. RESULTS: Temperature had a key role in increasing significantly the virucidal activity of VANTOCIL TG, reducing virus concentration by 4-log(10) within 10 minutes at 40 degrees C. The high temperature was linked to a reduction of viral aggregates despite the exposure to the biguanide. In addition, the viral inactivation kinetic became significantly more linear at 30 degrees C and 40 degrees C. Such results were also observed with sonication during treatment, where a first-order kinetic was observed. However, the addition of surfactants, even though there was evidence of a decrease in viral clumps, did not enhance the virucidal activity of polyhexamethylene biguanide. CONCLUSION: The presence of viral aggregates was an important factor in the virucidal efficacy of the biguanide as demonstrated by the correlation among high temperature, decrease in aggregates, and increase in activity, although it is possible that high temperatures might also cause conformational changes of the viral capsid, increasing the sensitivity of virions to the microbicide. PMID- 20006411 TI - Age-related effects on hippocampal precursor cell subpopulations and neurogenesis. AB - Hippocampal neurogenesis continuously declines in the aging brain but only little is known about age-related alterations in the subgranular zone (SGZ) of the dentate gyrus which accommodates different subpopulations of precursor cells. Here, we examined the age-related effects on total number and proliferation rate of distinct precursor cell populations in the dentate gyrus of 3 and 16 months old transgenic pNestin-GFP mice. Following a single injection of bromodeoxyuridine (BrdU) we observed a significant reduction of all proliferating precursor subtypes in aged mice compared to young controls. Stereological analysis further revealed that this decreased proliferation was not only caused by a general reduction in total number of precursor subtypes but also by a subtype-specific alteration of the proliferation rate. Whereas radial glia-like and early neuronal precursor cells demonstrate decreased proliferation rates, no difference was found for doublecortin-positive precursors. Additional long-term experiments further revealed that these age-related alterations in the proliferative zone were accompanied by a strongly decreased neurogenesis while hippocampal function was not impaired. PMID- 20006412 TI - [Transfusion in clinical practice in geriatric wards]. AB - PURPOSE: In the absence of specific recommendations on blood transfusion in elderly subjects, we carried out a survey to assess transfusion practices in geriatric medicine. METHODS: A descriptive, national, cross-sectional survey was conducted in 14 French geriatric departments (12 teaching hospitals and two general hospitals). In each department, five patients receiving transfusions were randomly selected in order to analyze their characteristics, the indications of blood transfusion, the criteria for and the methods of transfusion compared with Afssaps recommendations on transfusion thresholds. RESULTS: Data were analyzed for 70 patients (mean age 86+/-7 years, sex ratio female to male 1.8, with an average of five+/-two pathologies and six+/-three treatments). The indicators of poor tolerance included confusion (23 %), somnolence (22 %), acute heart failure (17 %) or coronary heart disease (16 %), and differed from the Afssaps criteria in the majority of cases. The transfusion threshold that were considered in the absence of poor tolerance (45 % of transfusions) differed from that recommended by Afssaps in 26 % of cases. The main adverse event in transfusion recipients was heart failure. CONCLUSION: When criteria for poor anaemia tolerance or transfusion thresholds are considered, transfusion practices in geriatric subjects have specific features. Further studies are needed to validate the appropriateness of the practices described in this survey. PMID- 20006413 TI - Outcomes of hemodialysis patients in a long-term care hospital setting: a single center study. AB - BACKGROUND: Long-term care hospitals (LTCHs) provide intermediary care after an acute-care hospitalization and usually furnish care to patients with complex medical problems. Outcomes of hemodialysis patients admitted to LTCHs, which includes patients with either end-stage renal disease (ESRD) or acute kidney injury (AKI) requiring dialysis therapy, are not known. STUDY DESIGN: Observational study. SETTING & PARTICIPANTS: All consecutive hemodialysis patients admitted to an LTCH. PREDICTORS: Demographic characteristics, comorbid and laboratory variables, ESRD, and AKI status during LTCH stay. OUTCOMES: Disposition from LTCHs was classified as discharge to home, nursing home, death in LTCH or hospice care, and re-admission to the hospital. In patients with AKI, renal recovery was defined as discontinuation of dialysis therapy before meeting disposition outcomes. RESULTS: 96 of 206 (46.6%) patients had ESRD, whereas 110 of 206 (53.3%) developed AKI requiring dialysis therapy during the acute-care hospitalization. 63 of 206 (31%) were discharged to home, 11 of 206 (5.4%) died or transferred to hospice, 81 of 206 (40%) went to a nursing home, and 49 of 206 (24%) were re-admitted to a hospital; mortality after re-admission was 32%. Older age (OR, 0.96; 95% CI, 0.93-0.98), diabetes mellitus (OR, 0.45; 95% CI, 0.23 0.94), number of re-admissions to the hospital (OR, 0.38; 95% CI, 0.18-0.78), aminoglycoside use (OR, 0.16; 95% CI, 0.04-0.64), and duration of hospitalization before LTCH admission (OR, 0.96; 95% CI, 0.94-0.99) were associated with lower odds of discharge to home. Of 110 patients with AKI requiring dialysis therapy, 30% (33 patients) discontinued dialysis therapy, whereas 70% were deemed to have ESRD on discharge. LIMITATIONS: Retrospective observational study. CONCLUSIONS: Most dialysis patients at LTCHs are either re-admitted to acute-care hospitals or require nursing home placement. Only 30% of patients with AKI recover sufficiently to discontinue dialysis therapy, whereas 70% are deemed to have ESRD. PMID- 20006415 TI - Perceived social position and health: Is there a reciprocal relationship? AB - Recent work exploring the relationship between socioeconomic status and health has employed a psychosocial concept called perceived social position as a predictor of health. Perceived social position is likely the "cognitive averaging" (Singh-Manoux, Marmot, & Adler, 2005) of socioeconomic characteristics over time and, like other socioeconomic factors, is subject to interplay with health over the life course. Based on the hypothesis that health can also affect perceived social position, in this paper we used structural equation modeling to examine whether perceived social position and three different health outcomes were reciprocally related in the Wisconsin Longitudinal Study, a longitudinal cohort study of older adults in the United States. The relationship between perceived social position and health differed across health outcomes-self reported health, the Health Utilities Index, and depressive symptoms-as well as across operationalization of perceived social position-compared to the population of the United States, compared to one's community, and a latent variable of which the two items are indicators. We found that perceived social position affected self-reported health when operationalized as latent and US perceived social position, yet there was a reciprocal relationship between self-reported health and community perceived social position. There was a reciprocal relationship between perceived social position and the Health Utilities Index, and depressive symptoms affected perceived social position for all operationalization of perceived social position. The findings suggest that the causal relationship hypothesized in prior studies--that perceived social position affects health- does not necessarily hold in empirical models of reciprocal relationships. Future research should interrogate the relationship between perceived social position and health rather than assume the direction of causality in their relationship. PMID- 20006416 TI - Adolescent brain development: current themes and future directions. Introduction to the special issue. PMID- 20006414 TI - A systematic review of therapeutic alliance, group cohesion, empathy, and goal consensus/collaboration in psychotherapeutic interventions in cancer: Uncommon factors? AB - The effects of four empirically supported therapeutic relationship factors (therapeutic alliance, empathy, goal consensus/collaboration, and group cohesion) on the outcome of psychotherapeutic interventions conducted with individuals living with cancer were systematically reviewed. PubMed, PsycINFO, and CINAHL were searched from their inception through November 13, 2008. Studies of psychotherapeutic interventions targeted to individuals living with cancer, which also empirically assessed the association between any of these therapeutic relationship factors and psychotherapy outcome were included in the review (8 of 742 papers initially reviewed). Information on study methodology and results were abstracted independently by the authors using a standardized form. Results indicated that therapist-rated rapport and group cohesion were significantly related to positive psychotherapeutic outcomes. No studies examined empathy. The literature on collaboration was mixed, but showed some support for increased collaboration being related to positive therapeutic outcomes. Overall the current literature on the role of therapeutic relationship factors in the context of individuals living with cancer is scant, and much more research is needed to determine the overall contribution of these four relationship elements to the outcomes of psychotherapeutic interventions for individuals living with cancer. Results of such studies could have important clinical and research implications. PMID- 20006417 TI - Outcome after resuscitation using controlled rapid extracorporeal cooling to a brain temperature of 30 degrees C, 24 degrees C and 18 degrees C during cardiac arrest in pigs. AB - AIM OF THE STUDY: To identify the optimal level of hypothermia during cardiac arrest, just prior to resuscitation with an extracorporeal cooling system and without fluid overload, for neurological outcome at day 9 in pigs. METHODS: In a prospective randomised laboratory investigation, 24 female Large White pigs (31 38 kg) underwent ventricular-fibrillation cardiac arrest for 15 min, followed by 1 min, 3 min or 5 min (n=8 per group) of 4 degrees C cooling with an extracorporeal cooling system via an aortic balloon catheter and resuscitation with cardiopulmonary bypass. Sixty minutes following induction of cardiac arrest, defibrillation attempts were started. Mild hypothermia (34.5 degrees C) and intensive care were continued for 20 h and final outcome was evaluated after 9 days. RESULTS: Brain temperature decreased from 38.5 degrees C to 30.4+/-1.6 degrees C within 221+/-81 s in the 1-min group; to 24.2+/-4.6 degrees C within 375+/-127 s in the 3-min group; and to 18.8+/-4.0 degrees C within 450+/-121 s in the 5-min group. Restoration of spontaneous circulation was achieved in seven (1 min group), six (3-min group) and six (5-min group) animals (p=0.78), whereas survival to 9 days was only achieved in six, three and three animals in each group (p=0.22), respectively. CONCLUSIONS: An extracorporeal cooling system rapidly induced brain hypothermia following prolonged normovolaemic cardiac arrest in pigs. Difference in outcome was not statistically significant amongst the three groups with various levels of hypothermia (30 degrees C, 24 degrees C and 18 degrees C) during cardiac arrest prior to resuscitation; however, the animals with the least temperature reduction showed a trend to better survival at 9 days. Further studies are necessary to investigate optimised methods for induction, as well as level, of cerebral hypothermia. PMID- 20006418 TI - Out-of-hospital airway management by paramedics and emergency physicians using laryngeal tubes. AB - CONCEPT: Endotracheal intubation (ETI) is considered to be the gold standard of prehospital airway management. However, ETI requires substantial technical skills and ongoing experience. Because failed prehospital ETI is common and associated with a higher mortality, reliable airway devices are needed to be used by rescuers less experienced in ETI. We prospectively evaluated the feasibility of laryngeal tubes used by paramedics and emergency physicians for out-of-hospital airway management. MATERIAL AND METHODS: During a 24-month period, all cases of prehospital use of the laryngeal tube disposable (LT-D) and laryngeal tube suction disposable (LTS-D) within five operational areas of emergency medical services were recorded by a standardised questionnaire. We determined indications for laryngeal tube use, placement success, number of placement attempts, placement time and personal level of experience. RESULTS: Of 157 prehospital intubation attempts with the LT-D/LTS-D, 152 (96.8%) were successfully performed by paramedics (n=70) or emergency physicians (n=87). The device was used as initial airway (n=87) or rescue device after failed ETI (n=70). The placement time was < or =45s (n=120), 46-90s (n=20) and >90s (n=7). In five cases the time needed was not specified. The number of placement attempts was one (n=123), two (n=25), three (n=2) and more than three (n=2). The majority of users (61.1%) were relative novices with no more than five previous laryngeal tube placements. CONCLUSION: The LT-D/LTS-D represents a reliable tool for prehospital airway management in the hands of both paramedics and emergency physicians. It can be used as an initial tool to secure the airway until ETI is prepared, as a definitive airway by rescuers less experienced with ETI or as a rescue device when ETI has failed. PMID- 20006419 TI - Opportunities for Emergency Medical Services care of sepsis. AB - OBJECTIVE: Emergency Medical Services (EMS) systems play key roles in the rapid identification and treatment of critical illness such as trauma, myocardial infarction and stroke. EMS often provides care for sepsis, a life-threatening sequelae of infection. In this study of Emergency Department patients admitted to the hospital with an infection, we characterized the patients receiving initial care by EMS. METHODS: We prospectively studied patients with suspected infection presenting to a 50,000 visit urban, academic ED from September 16, 2005-September 30, 2006. We included patients who had abnormal ED vital signs or required hospital admission. We identified patients that received EMS care. Between EMS and non-EMS patients, we compared patient age, sex, nursing home residency, vital signs, comorbidities, source of infection, organ dysfunction, sepsis severity and mortality. We analyzed the data using univariate odds ratios, the Wilcoxon rank sum test and multivariate logistic regression. RESULTS: Of 4613 ED patients presenting with serious infections, 1576 (34.2%) received initial EMS care. The mortality rate among those transported by EMS was 126/1576 (8.0%) compared to 67/3037 (2.2%) in those who were not. Adjusted mortality was higher for EMS (OR 1.8, 95% CI: 1.3-2.6). Of patients who qualified for protocolized sepsis care in the ED, 99/162 (61.1%) were transported via EMS. EMS patients were more likely to present with severe sepsis (OR 3.9; 3.4-4.5) or septic shock (OR 3.6; 2.6-5.0). EMS patients had higher sepsis acuity (mortality in ED sepsis score 6 vs. 3, p<0.001). CONCLUSIONS: EMS provides initial care for over one-third of ED infection patients, including the majority of patients with severe sepsis, septic shock, and those who ultimately die. EMS systems may offer important opportunities for advancing sepsis diagnosis and care. PMID- 20006420 TI - Total hip replacement at thirty years. Case report of crippling complications of a transient osteoporosis of the hip during pregnancy. PMID- 20006421 TI - Neuromuscular blockade after therapy with magnesium sulfate and amlodipine. PMID- 20006423 TI - Re: Markus J. Bader, Ronald Sroka, Christian Gratzke, et al. Laser therapy for upper urinary tract transitional cell carcinoma: indications and management. Eur Urol 2009;56:65-71. PMID- 20006425 TI - Mechanisms of breast cancer bone metastasis. AB - Bone, as well as liver and lung, is one of the most preferential metastatic target sites for cancers including breast, prostate, and lung cancers and the consequences are always devastating. Like other metastasis, breast cancer bone metastasis consists of several steps from the escape of primary site to the colonization in target site. This review focuses on several key steps including: 1. Invasion and escape from primary tumor site. 2. Target migration toward bone. 3. Specific adhesion and arrest in bone. 4. Establishment of metastasis in bone. The factors involved in this process will provide good targets for therapy. PMID- 20006424 TI - Unilateral deep brain stimulation of the nucleus accumbens in patients with treatment-resistant obsessive-compulsive disorder: Outcomes after one year. AB - OBJECTIVE: To investigate the effects of unilateral deep brain stimulation (DBS) in the right nucleus accumbens in patients with obsessive-compulsive disorder (OCD). Predominantly bilateral stimulation of the anterior limb of the internal capsule was utilized. METHODS: The study was designed as a double-blind sham controlled crossover study. Patients received 3 months of deep brain stimulation followed by 3 months of sham stimulation, or vice versa. Subsequently, stimulation was continued unblinded for all patients. The primary outcome measure was the severity level of OCD, measured using the Yale-Brown Obsessive Compulsive Scale (Y-BOCS). Secondary outcome measures were depressive symptoms, anxiety, psychological symptom severity, global functioning, quality of life, and cognitive function. RESULTS: The mean Y-BOCS scores decreased significantly from 32.2 (+/-4.0) at baseline to 25.4 (+/-6.7) after 12 months (p=0.012). Five out of ten patients showed a decrease of more than 25%, indicating at least a partial response. One patient showed a decrease in Y-BOCS severity greater than 35%. Similarly, depression, global functioning and quality of life improved within one year. In contrast, anxiety, global symptom severity and cognitive function showed no significant changes. In general, DBS was well-tolerated. CONCLUSIONS: DBS of the unilateral right nucleus accumbens showed encouraging results in patients with treatment-resistant OCD. Five out of ten patients reached at least a partial response after the first year. PMID- 20006426 TI - Simultaneous removal of 2,4-dichlorophenol and Cd from soils by electrokinetic remediation combined with activated bamboo charcoal. AB - An in situ electrokinetic remediation technique was designed by combining the uniform electrokinetic technology with a new-type of bamboo charcoal as adsorbent. A bench-scale experiment was conducted to investigate the application of this technique for simultaneous removal of 2,4-dichlorophenol (2,4-DCP) and Cd from a sandy loam at different periodic polarity-reversals. The contaminated soil was artificially spiked with 100 mg/kg 2,4-DCP and 500 mg/kg Cd. Two modes of polarity-reversal intervals of 12 and 24 h were included. After 10.5 d of operation, about 75.97% of Cd and 54.92% of 2,4-DCP were removed from soil at intervals of 24 h, whilst only 40.13% of Cd and 24.98% of 2,4-DCP were removed at intervals of 12 h. Soil water contents under two operation modes both significantly decreased, but evenly distributed spatially. Soil pH values under two operation modes were all maintained in the range from 7.2 to 7.4, close to the initial value. The electricity consumption per day was 12.24 and 11.61 kWh/m(3)/d, respectively at polarity-reversal intervals of 12 and 24 h. In conclusion, at polarity-reversal interval of 24 h, electroremediation combined with activated bamboo charcoal was effective in simultaneous removal of 2,4-DCP and Cd from soil. Our results indicate a promising potential in in situ electroremediation of soils co-contaminated with organics and heavy metals. PMID- 20006427 TI - Hazardous gas releases in urban areas: assessment of consequences through CFD modelling. AB - Release of hazardous materials in urban areas is a major concern in industrial risk assessment. The presence of high population density in such areas multiplies the magnitude of the consequences. In urban areas, many buildings with complex geometries are involved leading to 3D flow fields that strongly influence gas dispersion. Representing such complex geometries simply but realistically in detailed simulation models can be cumbersome and often limit their utility. In this work, a methodology for the construction of 3D urban models and their importation into CFD models was developed through the access to spatial geodatabases, leading to a relatively fast and simple domain design technique. Moreover, since the magnitude of consequences depends on the absorbed dose which in turn depends on both concentration and exposure time, a simple methodology for dose evaluation was developed and implemented in a CFD code that enables the estimation of regions with a given death probability. The approach was developed and applied to a case study with different atmospheric stratification conditions. The results were then compared with those obtained using integral models. It was found that integral models can both overestimate and underestimate the magnitude of consequences related to hazardous material releases in urban areas. PMID- 20006428 TI - Cr(VI) removal from aqueous solution by dried activated sludge biomass. AB - Batch experiments were conducted to remove Cr(VI) from aqueous solution using activated sludge biomass. The effects of acid pretreatment of the biomass, initial pH, biomass and Cr(VI) concentrations on Cr(VI) removal efficiency were investigated. Proton consumption during the removal process and the reducing capacity of sludge biomass were studied. The results show that acid pretreatment could significantly improve Cr(VI) removal efficiency and increase Cr(VI) reducing capacity by 20.4%. Cr(VI) removal was remarkably pH-dependent; lower pH (pH=1, 2) facilitated Cr(VI) reduction while higher pH (pH=3, 4) favored sorption of the converted Cr(III). Lower Cr(VI) concentration as well as higher biomass concentration could accelerate Cr(VI) removal. Cr(VI) reduction was not the only reason for proton consumption in the removal process. Pseudo-second-order adsorption kinetic model could successfully simulate Cr(VI) removal except under higher pH conditions (pH=3, 4). PMID- 20006429 TI - Electricity generation from indole and microbial community analysis in the microbial fuel cell. AB - Indole is a typical refractory and inhibitory compound present in coking wastewater. The aim of this study was to investigate possible electricity generation with indole degradation in the microbial fuel cell (MFC). Experiments were conducted in two types of the MFC: a continuous-fed MFC (C-MFC) and a batch fed MFC (B-MFC). In the C-MFC, the maximum power densities reached 45.4, 51.2, and 2.1 W/m(3), respectively, from using 1000 mg/L glucose, a mixture of 1000 mg/L glucose and 250 mg/L indole, and 250 mg/L indole as the fuel. When using 250 mg/L indole as the fuel, the removal efficiency of indole was up to 88% within 3 h. Increasing indole concentrations from 250 to 1500 mg/L resulted in decrease of the maximum power densities from 2.1 to 0.8 W/m(3), and average degradation rates from 41.7 to 8.9 mg/(Lh). Compared with the C-MFC, the B-MFC increased the maximum power densities from 2.1 to 3.3 W/m(3) and the coulombic efficiencies from 0.7% to 81.5%. Microbial community analyses showed that the addition of indole obviously changes the microbial community of the anode electrode, including the changes of relative abundance and emergence of new species. The results should be useful for treatment of wastewater containing indole. PMID- 20006430 TI - Research on the characteristics of red mud granular adsorbents (RMGA) for phosphate removal. AB - Red mud (RM), a waste tailing from alumina industry, was employed with bentonite and starch as the main raw materials for producing granular adsorbents in this study. The important parameters, which greatly affect the characteristics of red mud granular adsorbents (RMGA), such as the mass ratio of three raw materials, preheating time, preheating temperature, sintering time and sintering temperature (ST), were investigated. Adsorption capacities for various RMGA were described in terms of removal of phosphate from aqueous solution according to the adsorption experiments, in which certain operation parameters (like stirring speed, reaction time, adsorbent dosage, initial pH and initial phosphate concentration) at three different aquatic temperatures (AT) were applied. The results showed that the optimum ST, under which the largest adsorption capacity was achieved for RMGA with certain RM ratio, varied with different AT in adsorption process. Based on the further characterizations of RMGA, it was speculated that the performance above was caused by the competitive interactions of chemical adsorption and physical adsorption on RMGA, since the increase in ST could lead to qualitative diversification on the surface and quantitative decrease of effective components for adsorption simultaneously. PMID- 20006432 TI - A stochastic optimization model under modeling uncertainty and parameter certainty for groundwater remediation design--part I. Model development. AB - Solving groundwater remediation optimization problems based on proxy simulators can usually yield optimal solutions differing from the "true" ones of the problem. This study presents a new stochastic optimization model under modeling uncertainty and parameter certainty (SOMUM) and the associated solution method for simultaneously addressing modeling uncertainty associated with simulator residuals and optimizing groundwater remediation processes. This is a new attempt different from the previous modeling efforts. The previous ones focused on addressing uncertainty in physical parameters (i.e. soil porosity) while this one aims to deal with uncertainty in mathematical simulator (arising from model residuals). Compared to the existing modeling approaches (i.e. only parameter uncertainty is considered), the model has the advantages of providing mean variance analysis for contaminant concentrations, mitigating the effects of modeling uncertainties on optimal remediation strategies, offering confidence level of optimal remediation strategies to system designers, and reducing computational cost in optimization processes. PMID- 20006431 TI - Development of chitosan supported zirconium(IV) tungstophosphate composite for fluoride removal. AB - A new biocomposite was prepared by incorporating inorganic ion exchanger namely zirconium(IV) tungstophosphate (ZrWP) into the chitosan biopolymeric matrix. The sorption behaviour of fluoride from aqueous solutions by this ZrWP/chitosan (ZrWPCs) composite has been investigated by batch technique. The fluoride sorption was studied as a function of contact time, pH, initial fluoride concentration, competing co-ions and temperature. The defluoridation capacity (DC) of the adsorbent was found to be 2025 mg F(-) kg(-1). The composite was characterized using FTIR and SEM with EDAX analysis. The equilibrium sorption data were fitted to Freundlich and Langmuir isotherms. The kinetics of sorption was found to follow pseudo-second-order and intraparticle diffusion models. The values of thermodynamic parameters indicate the nature of sorption is spontaneous and endothermic. The biocomposite was successfully used for the removal of fluoride from the field water taken in a nearby fluoride endemic village. PMID- 20006433 TI - A stochastic optimization model under modeling uncertainty and parameter certainty for groundwater remediation design: part II. Model application. AB - A new stochastic optimization model under modeling uncertainty (SOMUM) and parameter certainty is applied to a practical site located in western Canada. Various groundwater remediation strategies under different significance levels are obtained from the SOMUM model. The impact of modeling uncertainty (proxy simulator residuals) on optimal remediation strategies is compared to that of parameter uncertainty (arising from physical properties). The results show that the increased remediation cost for mitigating modeling-uncertainty impact would be higher than those from models where the coefficient of variance of input parameters approximates to 40%. This provides new evidence that the modeling uncertainty in proxy-simulator residuals can hardly be ignored; there is thus a need of investigating and mitigating the impact of such uncertainties on groundwater remediation design. This work would be helpful for lowering the risk of system failure due to potential environmental-standard violation when determining optimal groundwater remediation strategies. PMID- 20006434 TI - Decontamination of soils containing PAHs by electroremediation: a review. AB - During the last years, the anthropogenic sources have contributed to organic compound penetration into the environment. One large group of persistent and toxic contaminants is the hydrophobic organic contaminants. Among them, polycyclic aromatic hydrocarbons (PAHs) have been recognized as a representative group of these pollutants with low solubility. In this paper, it is showed the electroremediation of soil contaminated with PAHs as an alternative, to organic compound removal. This technique, mainly used for heavy metal extraction, applies the electric current to promote the movement of contaminants. Nowadays the application of this technique alone or combined with other techniques as for example Fenton or bioremediation is taking fine results to PAHs removal. Although the PAHs soil decontamination by means of the electric field is in an initial stage, many researchers have demonstrated the treatment effectiveness. This paper describes the foremost principles to carry out the electroremediation of soils contaminated with PAHs, just like the different alternatives to improve the electroremediation of PAHs and also the new methodologies of PAHs removal by using hybrid technologies. PMID- 20006435 TI - Application of vegetable oils in the treatment of polycyclic aromatic hydrocarbons-contaminated soils. AB - A brief review is conducted on the application of vegetable oils in the treatment of PAH-contaminated soils. Three main scopes of treatment strategies are discussed in this work including soil washing by oil, integrated oil-biological treatment and integrated oil-non-biological treatment. For each of these, the arguments supporting vegetable oil application, the applied treatment techniques and their efficiencies, associated factors, as well as the feasibility of the techniques are detailed. Additionally, oil regeneration, the environmental impacts of oil residues in soil and comparison with other commonly employed techniques are also discussed. PMID- 20006436 TI - Determination of ultra-trace aluminum in human albumin by cloud point extraction and graphite furnace atomic absorption spectrometry. AB - A cloud point extraction (CPE) method for the preconcentration of ultra-trace aluminum in human albumin prior to its determination by graphite furnace atomic absorption spectrometry (GFAAS) had been developed in this paper. The CPE method was based on the complex of Al(III) with 1-(2-pyridylazo)-2-naphthol (PAN) and Triton X-114 was used as non-ionic surfactant. The main factors affecting cloud point extraction efficiency, such as pH of solution, concentration and kind of complexing agent, concentration of non-ionic surfactant, equilibration temperature and time, were investigated in detail. An enrichment factor of 34.8 was obtained for the preconcentration of Al(III) with 10 mL solution. Under the optimal conditions, the detection limit of Al(III) was 0.06 ng mL(-1). The relative standard deviation (n=7) of sample was 3.6%, values of recovery of aluminum were changed from 92.3% to 94.7% for three samples. This method is simple, accurate, sensitive and can be applied to the determination of ultra trace aluminum in human albumin. PMID- 20006437 TI - Development of an on-line temperature-assisted ionic liquid dispersive microextraction system for sensitive determination of vanadium in environmental and biological samples. AB - An original flow injection (FI) system was developed for on-line microextraction of Vanadium (V) based on room temperature ionic liquid (RTIL). Vanadium was complexed with 2-(5-bromo-2-pyridylazo)-5-diethylaminophenol (5-Br-PADAP) at pH 4.0. A 40 microL-volume of 1-butyl-3-methylimidazolium hexafluorophosphate ([C(4)mim][PF(6)]) RTIL was mixed with 5 mL of sample solution containing the V-5 Br-PADAP complex. Then, a fully on-line temperature-assisted dispersion procedure was developed, followed by, analyte microextraction; and final on-line separation of the RTIL phase with a florisil-containing microcolumn. Vanadium was removed from the microcolumn with a 10% (v/v) nitric acid (in acetone) solution, and finally measured by electrothermal atomic absorption spectrometry (ETAAS). The detection limit achieved after preconcentration of 5 mL of sample solution, was 4.8 ng L(-1). The relative standard deviation (RSD) for 10 replicate determinations at 5 microg L(-1) of vanadium level was 4.1%, calculated from the obtained peak heights. The calibration graph was linear, with a correlation coefficient of 0.9982 at levels from the detection limits up to 15 microg L(-1). The method was successfully applied for the determination of vanadium in environmental and biological samples. PMID- 20006438 TI - Physical and biological treatments of polyethylene-rice starch plastic films. AB - This study aimed to produce an industrial applicable thermo-stable alpha-amylase from marine Bacillus amyloliquefaciens which isolated and selected according to its significant enzyme production. The effect of different pH values and temperatures on the bacterial growth and the enzyme production was estimated using an experimental statistical design; maximum amylase production and bacterial growth was obtained at pH 7.0 and 50 degrees C. Some biodegradable polyethylene rice starch plastic films (PERS-P) were manufactured using 0, 2.5, 5, 7.5 and 10% starch concentrations. The biodegradability (reduction in the plastic elongation%) was tested using the exposure to UV radiation at lambda(300 400 nm) (intensity of about 1000 W/m(2)) and the produced B. amyloliquefaciens thermo-stable alpha-amylase. A significant reduction in the elongation% of these biodegradable plastics was observed in both cases especially on testing the 10% PERS-P; they showed a reduction of 26% and 20%, respectively, compared to the untreated plastic films (180+/-5). PMID- 20006439 TI - Effects of uniformities of deposition of respirable particles on filters on determining their quartz contents by using the direct on-filter X-ray diffraction (DOF XRD) method. AB - In this study, field samplings were conducted in three workplaces of a foundry plant, including the molding, demolding, and bead blasting, respectively. Three respirable aerosol samplers (including a 25-mm aluminum cyclone, nylon cyclone, and IOSH cyclone) were used side-by-side to collect samples from each selected workplace. For each collected sample, the uniformity of the deposition of respirable dusts on the filter was measured and its free silica content was determined by both the DOF XRD method and NIOSH 7500 XRD method (i.e., the reference method). A same trend in measured uniformities can be found in all selected workplaces: 25-mm aluminum cyclone>nylon cyclone>IOSH cyclone. Even for samples collected by the sampler with the highest uniformity (i.e., 25-mm aluminum cyclone), the use of the DOF XRD method would lead to the measured free silica concentrations 1.15-2.89 times in magnitude higher than that of the reference method. A new filter holder should be developed with the minimum uniformity comparable to that of NIOSH 7500 XRD method (=0.78) in the future. The use of conversion factors for correcting quartz concentrations obtained from the DOF XRD method based on the measured uniformities could be suitable for the foundry industry at this stage. PMID- 20006440 TI - Scanning moire fringe imaging by scanning transmission electron microscopy. AB - A type of artificial contrast found in annular dark-field imaging is generated by spatial interference between the scanning grating of the electron beam and the specimen atomic lattice. The contrast is analogous to moire fringes observed in conventional transmission electron microscopy. We propose using this scanning interference for retrieving information about the atomic lattice structure at medium magnifications. Compared with the STEM atomic imaging at high magnifications, this approach might have several advantages including easy observation of lattice discontinuities and reduction of image degradation from carbon contamination and beam damage. Application of the technique to reveal the Burgers vector of misfit dislocations at the interface of epitaxial films is demonstrated and its potential for studying strain fields is discussed. PMID- 20006441 TI - A framework for automatic segmentation in three dimensions of microstructural tomography data. AB - Routine use of quantitative three dimensional analysis of material microstructure by in particular, focused ion beam (FIB) serial sectioning is generally restricted by the time consuming task of manually delineating structures within each image slice or the quality of manual and automatic segmentation schemes. We present here a framework for performing automatic segmentation of complex microstructures using a level set method. The technique is based on numerical approximations to partial differential equations to evolve a 3D surface to capture the phase boundaries. Vector fields derived from the experimentally acquired data are used as the driving forces. The framework performs the segmentation in 3D rather than on a slice by slice basis. It naturally supplies sub-voxel precision of segmented surfaces and allows constraints on the surface curvature to enforce a smooth surface in the segmentation. Two applications of the framework are illustrated using solid oxide cell materials as examples. PMID- 20006442 TI - The life cycle and biological characteristics of Dermacentor silvarum Olenev (Acari: Ixodidae) under field conditions. AB - In this study, we investigated the life cycle and biological characteristics of all developmental stages of Dermacentor silvarum under field conditions in Hebei province in North China. Larvae, nymphs and adults were fed on rabbits, and maintained under natural lighting and climate conditions. Ticks were released into a confined plot located in natural tick habitat, and we observed the host questing behavior and seasonal occurrence. Due to a behavioral diapause in females and the synchronization of oviposition and larval hatching, D. silvarum only active from late February to early September under field conditions. The duration of the life cycle of D. silvarum was highly variable due to different preovipositional and egg hatching periods for ticks engorging in March, April or May. The developmental periods of larvae (18 days) and nymphs (29 days) under field conditions were similar to those under laboratory conditions. Moreover, there was a highly significant correlation between engorged body weight of females and egg masses laid (r=0.76, p<0.001). The female reproductive efficiency index (REI) in March, April and May was 5.7, 7.1 and 6.2, respectively. Finally, we also observed different ovipositional patterns between females that engorged in the field and those that engorged under laboratory conditions. PMID- 20006443 TI - Microsporidia in exotic birds: intermittent spore excretion of Encephalitozoon spp. in naturally infected budgerigars (Melopsittacus undulatus). AB - Nine naturally infected asymptomatic budgerigars (Melopsittacus undulatus) were screened daily for microsporidia spore excretion during a 30-day period and the localization of infection was evaluated using microscopy and molecular methods. While the microscopic examination revealed 2.4% positivity out of all fecal samples, using PCR the positivity was 10x higher (24.6%). All nine budgerigars excreted microsporidial spores intermittently in irregular intervals with 1-11 day long interruptions. Most of the birds were infected simultaneously with Encephalitozoon cuniculi and Encephalitozoon hellem. While histological and TEM examination failed to confirm the presence of microsporidial spores in tissues, the PCR detected microsporidial DNA mostly in the small intestine, liver and lungs of four selected budgerigars dissected. Despite the chronic infection proved using molecular methods, no clinical signs of disease were observed during monitoring and no pathological findings were found during dissection. PMID- 20006444 TI - Prevention of Ichthyophthirius multifiliis infestation in goldfish (Carassius auratus) by potassium ferrate(VI) treatment. AB - Ichthyophthirius multifiliis is an important freshwater teleost pathogen that often leads to significant economic losses to the aquaculture industry. The purpose of this study was to assess the acute toxicity of potassium ferrate(VI) to I. multifiliis theront and the concentration needed to prevent I. multifiliis infestation in goldfish, Carassius auratus. Five hundred theronts were exposed to concentrations of potassium ferrate(VI) in each well of a 96-well microtiter plate and observed for 4h to determine the acute toxicity. Results showed that the exposure of I. multifiliis theronts to potassium ferrate(VI) at concentrations of 4.80mg/L or more resulted in 100% mortality by 4h; the LC(50) value was estimated to be 1.71mg/L. Aqueous static renewal 96-h bioassays were carried out to determine the acute toxicity of potassium ferrate(VI) to goldfish. The LC(50) value for potassium ferrate(VI) in goldfish was 42.51mg/L. Goldfish were exposed to 4000 theronts/fish in aerated tap water (a dose previously shown to result in consistent infestation) and treated with a single dose of potassium ferrate(VI) after 30min contact with theronts. Infection level and prevalence were recorded everyday after exposure. The results revealed that potassium ferrate(VI) at the 4.80mg/L or more concentrations can significantly reduce not only the number of trophonts on the fin of goldfish on day 3 (P<0.05), but also the prevalence of ichthyophthiriasis (P<0.05). Potassium ferrate(VI) at a concentration of 4.80mg/L was considered to be the lowest effective dose to prevent infestation of I. multifiliis in goldfish. PMID- 20006445 TI - Assessing the severity of inhalation injuries in adults. AB - Inhalation injury is a major cause of morbidity and mortality in thermally injured patients but is difficult to quantify. A number of parameters can be useful evaluating inhalation injury including bronchoscopy, PaO(2)/FiO(2) (P/F) ratio, and carboxyhaemaglobin (COHb) levels. Identification of markers that permit early detection of the degree of damage caused by inhalation injury is important to predict patient survival. In this retrospective study we evaluated the potential of using the P/F ratio, bronchoscopic findings and the COHb level to assess the severity of the inhalation injury in adults. A retrospective review of all patients admitted with inhalation injuries to the intensive care unit (ICU) from January 1995 to December 2002 was conducted and of the 110 patients 105 Casenotes were obtained and investigated. The total burn surface area (TBSA), age and bronchoscopic findings showed a positive correlation with mortality. There was no significant correlation between COHb levels and mortality, but there was a significant correlation between P/F ratio and mortality in adults. Logistic regression indicated that the combination of TBSA, age and P/F ratio was highly predictive of mortality following major burns. This study supports the predictive nature of the P/F ratio in relation to survival in adults. Larger prospective studies using bronchoscopic findings and P/F ratio would give us a better indication as to the value of a scoring system in adults that are admitted with inhalation injuries. PMID- 20006446 TI - The involvement of alpha-melanocyte-stimulating hormone in the hyperpigmentation of human skin autografts. AB - Hyperpigmentation occurs frequently in human skin autografts, resulting in an unsatisfactory appearance. OBJECTIVE: This study aims to elucidate the role of alpha-melanocyte-stimulating hormone (alpha-MSH) in the hyperpigmentation process of skin autografts by analysing the expression of alpha-MSH and the amount of melanin in human autografted skin and normal skin. METHODS: Immunohistochemistry and reverse transcription-polymerase chain reaction methods were carried out to detect the expression and distribution of alpha-MSH and pro-opiomelanocortin (POMC) in skin autografts, including full-thickness skin autografts, split thickness skin autografts and normal skin. The Fontana-Masson stain was used to detect the amount of melanin in all the skin specimens. RESULTS: The level of alpha-MSH and POMC expression in autografted skin was much higher than that of control normal skin, and thinner split-thickness skin autografts expressed higher levels of POMC mRNA and alpha-MSH protein. The amount of melanin in skin autografts was significantly higher compared with normal skin. The expression of alpha-MSH correlated well with the amount of melanin in skin autograft epidermis. CONCLUSIONS: These results indicate that the function of melanogenesis, transfer of melanin in melanocytes, was dramatically enhanced in skin autografts by alpha MSH. The over-expression of alpha-MSH may play an important role in the hyperpigmentation process of skin autografts and provide a novel mechanism of this process. PMID- 20006447 TI - Socioeconomic deprivation and burns. AB - Burns are a complex phenomenon with multiple interactive factors. Age, ethnicity and socioeconomic deprivation have been identified in the literature as important determinants of this form of injury. OBJECTIVE: The primary objective of this study was to undertake a descriptive analysis of the socioeconomic factors associated with burn injuries in New Zealand. This analysis will help to identify those individuals at greatest risk of burn and assist with the design of targeted interventions that are more likely to be effective. METHOD: A retrospective review of the medical records of patients admitted acutely to hospital in New Zealand with a burn between 1996 and 2006. We reviewed the records with regards to patient demographics and socioeconomic position. RESULTS: A total of 14,708 admissions were reviewed. Stratification by age indicated that admissions were highest in the 0-4-year age group. The rate of admission was highest in the Maori ethnic group. The rate of admissions increased in proportion with increasing deprivation. CONCLUSION: This study confirms marked ethnic and socioeconomic disparities in burn injuries requiring hospital admission. This is unacceptable and preventable. We propose active targeting of burn prevention strategies at high-risk groups as a cost-effective way of reducing disparities. PMID- 20006449 TI - Pandemic influenza A (H1N1): mandatory vitamin D supplementation? PMID- 20006448 TI - Levels of maternal serum corticotropin-releasing hormone (CRH) at midpregnancy in relation to maternal characteristics. AB - BACKGROUND: Corticotropin-releasing hormone (CRH) in maternal blood originates primarily from gestational tissues and elevated levels in midpregnancy have been linked to adverse pregnancy outcomes. Investigators have hypothesized that high levels of maternal stress might lead to elevated CRH levels in pregnancy. Yet a few studies have measured maternal CRH levels among subgroups of women who experience disproportionate socioeconomic disadvantage, such as African-American and Hispanic women, and found that these groups have lower CRH levels in pregnancy. Our goal was to identify maternal characteristics related to CRH levels in midpregnancy and examine which if any of these factors help to explain race differences in CRH levels. METHODS: The Pregnancy Outcomes and Community Health (POUCH) Study prospectively enrolled women at 15-27 weeks' gestation from 52 clinics in five Michigan communities (1998-2004). Data from the POUCH Study were used to examine maternal demographics, anthropometrics, health behaviors, and psychosocial factors (independent variables) in relation to midpregnancy blood CRH levels modeled as logCRHpg/ml (dependent variable). Analyses were conducted within a sub-cohort from the POUCH Study (671 non-Hispanic Whites, 545 African-Americans) and repeated in the sub-cohort subset with uncomplicated pregnancies (n=746). Blood levels of CRH and independent variables were ascertained at the time of enrollment. All regression models included week of enrollment as a covariate. In addition, final multivariate regression models alternately incorporated different psychosocial measures along with maternal demographics and weight. Psychosocial variables included measures of current depressive symptoms, perceived stress, coping style, hostility, mastery, anomie, and a chronic stressor (history of abuse as a child and adult). RESULTS: In sub cohort models, the adjusted mean log CRH level was significantly lower in African Americans vs. non-Hispanic Whites; the difference was -0.48pg/ml (P<0.01). This difference was reduced by 21% (-0.38pg/ml, P<0.01) after inclusion of other relevant covariates. Adjusted mean log CRH levels were also lower among women with <12 years vs. >or=12 years of education (minimal difference=-0.19pg/ml, P<0.05), and among women with high levels of depressive symptoms who did not use antidepressants vs. women with lower levels of depressive symptoms and no antidepressant use (minimal difference=-0.13pg/ml, P<0.01). Log CRH levels were inversely associated with maternal weight (-0.03pg/ml per 10 pound increase, P<.05) but unrelated to smoking and all other psychosocial measures. Results were similar in the subset of women with uncomplicated pregnancies, except that lower CRH levels were also linked to higher perceived stress. CONCLUSION: African American women have lower blood CRH levels at midpregnancy and the race difference in CRH levels is reduced modestly after adjustment for other maternal characteristics. CRH levels were not elevated among women with high levels of perceived stress or more chronic stressors. The inverse association between CRH levels and maternal weight is likely due to a hemodilution effect. Relations among maternal CRH levels and maternal race, educational level, and depressive symptoms are difficult to explain and invite further investigation. Our results highlight a group of covariates that merit consideration in studies that address CRH in the context of pregnancy and/or post-partum complications. PMID- 20006450 TI - Metastasis: inherent vs. acquired phenotype. AB - The problem how tumor cells get the metastatic ability is still a hot debate. Based on the premise that the default state of normal cells is quiescent rather than mobile, the classic progression and early metastasis model suggested that tumor metastasis should be an acquired trait contributed by the late or early gene mutations during carcinogenesis. Here, an inherent metastasis model is proposed that the metastatic ability of the tumor cells is one of the constitutive features of the normal cells of tumor origin. The idea is based on two facts. One is that tumor arises from stem or progenitor cells and in turn are driven by tumor stem cells. The other is that emerging evidence showing that a small population of stem or progenitor cells has the inherent migration capacity in normal development and adulthood. This inherent metastatic model has some implications. First, metastatic dissemination should occur continually throughout the course of primary tumor development and generate a diverse spectrum of disseminated cells. Second, most of the disseminated tumor cells should have the stem cell like features. Third, migration of stem cells and cancer cells should invoke similar molecular processes involving metastasis. Fourth, genomic alterations that primarily promote the production of tumor cells with stem cell traits, i.e., tumor stem cell, exacerbate tumor progression and metastasis. Finally, overt metastatic production is primarily determined by whether the disseminated tumor cells can survive and grow into overt metastatic foci at the ectopic sites, instead of that whether the tumor cells can leave the primary sites and travel to other sites. All these predictions have gotten increasing supporting evidences. Yet, to confirm whether the new paradigm is true or false, it needs carefully examine whether normal stem or progenitor cells of various tissues have the potential to traveling and arriving at the ectopic sites. Furthermore, exploiting the mechanisms for regulating normal stem or progenitor cells migration may provide more critical results for our deeply understanding the secrets of tumor metastasis and offer new methods for preventing and treating tumor metastasis. PMID- 20006452 TI - Occurrence of Mycoplasma suis in wild boars (Sus scrofa L.). AB - Porcine infectious anemia is a well-known disease that occurs worldwide and is caused by the uncultivable hemotrophic bacterium Mycoplasma suis. In this study the occurrence of M. suis in wild boars was investigated by employing a quantitative real-time LightCycler PCR. M. suis infections were detected in 36 out of 359 wild boars (10.03%). Sequencing of the 16S rRNA gene and subsequent phylogenetic analysis revealed the existence of two genetically distinct M. suis subtypes in the wild boar population: one subtype was >99.0% identical to known American and European M. suis isolates, and the second subtype showed the highest homology to known Chinese isolates. In summary, this is the first detection of M. suis in wild boars. The role of M. suis as pathogen in wild boars has yet to be established, but the present findings revealed a possible wildlife reservoir for these bacteria. PMID- 20006451 TI - Nicotine withdrawal in U.S. smokers with current mood, anxiety, alcohol use, and substance use disorders. AB - BACKGROUND: The current study examined tobacco withdrawal symptoms and withdrawal related discomfort and relapse in smokers with and without current mood disorders, anxiety disorders, alcohol use disorders (AUD), and substance use disorders (SUD). METHODS: The subsample of current daily smokers (n=8213) from the National Epidemiologic Survey on Alcohol and Related Conditions (NESARC, Wave 1, 2001-2002, full sample n=43,093) were included in these analyses. Cross sectional data compared smokers with and without current psychiatric disorders on withdrawal symptoms using logistic regression models. The effects of having a co morbid psychiatric disorder and AUD/SUD compared to a psychiatric disorder alone on nicotine withdrawal were also examined. RESULTS: Participants with a current mood disorder, anxiety disorder, AUD, or SUD were more likely to report withdrawal symptoms and reported more withdrawal symptoms than those without current disorders. Having a current mood disorder, anxiety disorder, or SUD was also associated with increased likelihood of withdrawal-related discomfort and relapse. There were no significant interactions between psychiatric disorders and AUDs/SUDs on withdrawal symptoms or behavior. CONCLUSIONS: Participants with a current Axis I disorder were more likely to experience tobacco withdrawal symptoms and withdrawal-related discomfort and relapse. Having a co-morbid psychiatric disorder and AUD/SUD did not synergistically increase the experience of withdrawal-related symptoms or relapse. It is important to identify Axis I disorders in smokers and provide these smokers with more intensive and/or longer treatments to help them cope with withdrawal symptoms and prevent relapse. PMID- 20006453 TI - Buprenorphine and major metabolites in blood specimens collected for drug analysis in law enforcement purposes. AB - A liquid chromatographic/electrospray ionization tandem mass spectrometric method for the quantification of buprenorphine (BUP), norbuprenorphine (NBUP), buprenorphine-3-beta-D-glucuronide (BUPG) and norbuprenorphine-3-beta-D glucuronide (NBUPG) in serum samples was developed and validated. Pre-treatment of BUP and NBUP was by liquid-liquid extraction, while glucuronides were favourably isolated by solid phase extraction. Separation in 2 separate runs (2 x 5 min) was achieved using isocratic elution. The method was applied to 20 authentic serum specimens collected for law enforcement purposes where BUP intake had been indicated. The parent drug was not detectable in half of the specimens at a lower limit of detection of 0.2 ng/mL, whereas NBUP could be determined from any sample but one. NBUPG is the major metabolite present, which could be identified along with BUPG in all samples under investigation. In authentic specimens it could be advisable to monitor BUP metabolites along with the parent drug. PMID- 20006454 TI - MR pelvimetry measurements, analysis of inter- and intra-observer variation. AB - OBJECTIVE: To set reference standards and then to evaluate the measurement variation in magnetic resonance (MR) pelvimetry between observations and observers. METHODS: The study was carried out retrospectively using 100 MR pelvimetry examinations performed in North Carelian Central Hospital between September 2006 and January 2008. Pelvimetric parameters of pelvic inlet and outlet were measured four times to determine the standard reference for each measurement and then intra- and inter-observer variations were compared. RESULTS: The accuracy of MR pelvimetry, defined as a deviation of <5mm from the reference standard, was better in pelvic inlet measurements than in the corresponding outlet measurements (95-99% vs. 86-89%). Intra-observer variation was acceptable in all of the measurements with intraclass correlation coefficient (ICC) in the range 0.956-0.981 in all parameters. Inter-observer variation was higher than intra-observer variation. The largest variation of measurements was in pelvic outlet parameters between observers with the ICC in the range of 0.710-0.813. CONCLUSION: MR pelvimetry measurement should be conducted in a centralized location to decrease observer-related variation. Clinicians should be aware that millimeter differences are not reliable in MR pelvimetry and therefore the use of millimeter accurate limits are not recommended in obstetric decision making. PMID- 20006456 TI - Small and large bowel volvulus: Clues to early recognition and complications. AB - Small and large bowel volvulus are uncommon causes of bowel obstruction with nonspecific clinical manifestations which may delay the diagnosis and increase morbidity. Therefore, radiologists play an important role in promptly establishing the diagnosis, recognizing underlying congenital or acquired risk factors and detecting potentially life-threatening complications. Multidetector CT performed with intravenous contrast is currently the preferred modality for the evaluation of volvulus, which is best appreciated when imaging is perpendicular to the axis of bowel rotation, hence the benefit of multiplanar reformations. In this pictorial essay we review the pathophysiology of the different types of intestinal volvulus, discuss diagnostic criteria for prompt diagnosis of volvulus and emphasize early recognition of the complications. PMID- 20006455 TI - Efficacy and safety of lanthanoids as X-ray contrast agents. AB - OBJECTIVE: It has been suggested that elements from the lanthanoid (Ln) series may be well suited for use as absorbing elements in X-ray contrast agents (CA). Because gadolinium, an element of the lanthanoid series, has been identified as being possibly associated with nephrogenic systemic fibrosis (NSF), a rare but potentially severe disease, we sought to determine if other lanthanoids might possess a similar potential. MATERIALS AND METHODS: By computed tomography (CT), we compared the X-ray attenuation of all lanthanoids to that of iodine in vitro. In addition, we injected Han-Wistar rats on five consecutive days with 2.5 mmol Ln/kg bodyweight intravenously to test several Ln-DTPA-BMA complexes (praseodymium, europium, gadolinium, and holmium). Saline solution and a Ca-DTPA BMA group served as controls. Ln concentrations in the skin and organs were determined by inductively coupled plasma mass spectrometry (ICP-MS). This method measures the total Ln content and cannot differentiate between chelated and unchelated Ln. In addition, serum cytokine levels were measured by Luminex technology. The complex stability of the Ln-DTPA-BMA complexes was also assessed in vitro. RESULTS: Lanthanoids showed up to 50% higher X-ray attenuation than iodine in CT. The highest X-ray attenuation was observed with holmium and erbium. Differences in the in vitro complex stability of Pr-, Eu-, Gd-, and Ho-DTPA-BMA complexes were observed. The complex stability differences were also reflected by differences in the concentrations in tissue of the lanthanoids in vivo. Injections of Ln complexes caused NSF-like skin lesions in rats and a rapid upregulation of pro-fibrotic and inflammatory serum cytokines. The Ca-DTPA-BMA complex did not to induce pro-fibrotic cytokines or skin lesions. Pr-DTPA-BMA appeared to be toxic; all Pr-DTPA-BMA treated animals died within the first four days of the experiment and were therefore excluded from further analyses. CONCLUSION: Lanthanoids are very well suited for higher X-ray tube voltages, particularly CT examinations. However, Ln-specific induction of NSF-like skin lesions and rapid elevation of pro-fibrotic serum cytokines levels were observed in rats following multiple administrations of high doses of Ln-DTPA-BMA complexes. The results of this animal study suggest that the stability of lanthanoid complexes may be an important consideration in evaluating the potential for in vivo safety. Furthermore the results suggest a potential of the entire class of lanthanoids to have the potential to trigger NSF-like skin lesions in rats rather than only some of the specific elements of this series. PMID- 20006457 TI - Medical nutrition therapy for overweight youth in their medical home: the KIDPOWER experience. AB - OBJECTIVE: To determine the feasibility and impact of implementing a standardized medical nutrition therapy (MNT) protocol to treat overweight children in their medical home. Primary outcomes evaluated were changes in eating behaviors, television habits, and weight status. METHODS: A MNT protocol with handouts (known as KIDPOWER) was delivered by a registered dietitian (RD) to overweight children (2-20 years) in 9 primary care practices in a rural community. A medical record review yielded height, weight and selected patient/parent reported eating and TV behaviors from each MNT visit. Mean values were compared by paired samples t-tests. Changes in behaviors and BMI z score were compared using repeated measures ANOVA. RESULTS: For patients with at least three MNT visits (n=109) significant improvements in eating and TV viewing habits were seen by the third visit. Weight status, as determined by a significant decline in mean BMI z score, was improved. CONCLUSION: The KIDPOWER protocol delivered by a RD in the pediatric medical home aided overweight youth in making behavior changes that positively impacted their weight status. PRACTICE IMPLICATIONS: Reimbursement to RDs for treating childhood obesity is improving and KIDPOWER provides a model and tools for community practice. PMID- 20006458 TI - The realisation of patient-centred care during a 3-year proactive telephone counselling self-care intervention for diabetes. AB - OBJECTIVE: To explore the way that patient-centred care is realised within a tele carer behavioural change intervention. METHOD: In-depth, semi-structured interviews undertaken at years 1 and 3 with a purposively selected sample from the intervention group within a 3-year randomised controlled trial (RCT) of a telephone-based education and support for persons with type 2 diabetes, and interviews with the non-medically trained tele-carers and supervising diabetes specialist nurse. RESULTS: A four-phased flow of the patient-centred interactions was identified, which evolved over the process of the intervention. Initially, attention centred on building a picture for and of the patient and assessing their knowledge base. Later, focus moved towards understanding diabetes from the patient's perspective and advice-giving became more individualised. Throughout, the interaction dynamics varied for patients. CONCLUSION: This study provides insight into the development of patient-centred behaviours over time and the influence of patients on tele-carer communication styles. PRACTICE IMPLICATION: When adopting a patient-centred approach, tele-carers need to be flexible and recognise that patients vary in their knowledge, skills and psychological adaption to diabetes. Continuity of care and consistent contact is pivotal to patients being able to move through the various phases of their illness trajectory and make the transition towards improved self-care management. PMID- 20006459 TI - Patients' experiences with navigation for cancer care. AB - OBJECTIVE: We examined how navigation, defined as the assessment and alleviation of barriers to adequate health care, influences patients' perspectives on the quality of their cancer care. METHODS: We conducted post-study patient interviews from a randomized controlled trial (usual care vs. patient navigation services) from cancer diagnosis through treatment completion. Patients were recruited from 11 primary care, hospital and community oncology practices in New York. We interviewed patients about their expectations and experience of patient navigation or, for non-navigated patients, other sources of assistance. RESULTS: Thirty-five patients newly diagnosed with breast or colorectal cancer. Valued aspects of navigation included emotional support, assistance with information needs and problem-solving, and logistical coordination of cancer care. Unmet cancer care needs expressed by patients randomized to usual care consisted of lack of assistance or support with childcare, household responsibilities, coordination of care, and emotional support. CONCLUSION: Cancer patients value navigation. Instrumental benefits were the most important expectations for navigation from navigated and non-navigated patients. Navigated patients received emotional support and assistance with information needs, problem-solving, and logistical aspects of cancer care coordination. PRACTICE IMPLICATIONS: Navigation services may help improve cancer care outcomes important to patients by addressing fragmented, confusing, uncoordinated, or inefficient care. PMID- 20006460 TI - Themes affecting health-care consumers' choice of a hospital for elective surgery when receiving web-based comparative consumer information. AB - OBJECTIVE: To get insights into the decision-making strategy of health-care consumers when confronted with comparative consumer information. METHODS: Qualitative descriptive study among 18 consumers who had a hip or knee replacement no longer than five years ago. To study their decision-making strategies a paper draft for a website was used providing comparative consumer information. Data were collected by cognitive interviews and focus-group meetings and subjected to thematic analysis. RESULTS: Consumers were able to understand the presented information, but had problems to use it as a decision aid. They primarily relied on previous experiences. Four themes were revealed: decision making, perceived benefits, unmet information needs, and trustworthiness. Consumers used different decision strategies and showed unpredictable behavior when choosing a hospital. CONCLUSION: Individual decision strategies, unsatisfied information needs, limited tenability and too coarse aggregation levels of quality scores are barriers for a proper use of comparative consumer information. Personal experience remains a valuable information source for hospital selection. We suggest that a website presenting comparative consumer information should be flexible in various ways and should include functionality to share personal experience. PMID- 20006461 TI - Obese young people's accounts of intervention impact. AB - OBJECTIVE: To uncover in-depth qualitative accounts of intervention impact from obese young people during a period of lifestyle change after attending a residential weight-loss camp. METHODS: An innovative, progressively focused process of (1) single end-of-stay interviews, (2) interviews repeated in the home environment at 3 months and (3) 9 months. Comprehensive data were collected from five information-rich cases who contributed at each stage. Verbatim transcripts were analysed to identify issues associated with camp impact. RESULTS: Reporting positive in-camp experiences, there were also anxieties about returning home and successfully translating knowledge and behaviors into longer term strategies. Inductive analysis suggested cognitive ambiguity in relation to positive camp impact. This developed and intensified over the 9 months of post-camp experience, despite the help of supportive others. CONCLUSION: Young people described a positive post-camp impact that continued into the home environment. Unexpectedly, though it was also an ambiguous experience. Cognitive ambiguity created behavioral conflict which undermined motivation for sustained healthy living. PRACTICE IMPLICATIONS: These unique accounts enable service providers to better understand post-camp experiences and use them to work with young people towards more positive outcomes. Even after intensive and successful intervention young people may still require ongoing support for continued lifestyle change. PMID- 20006462 TI - Thyroid blood flow as a useful predictor of relapse of Graves' disease after normal delivery in patients with Graves' disease. AB - OBJECTIVE: Measurement of the peak systolic velocity (PSV) in the inferior thyroid artery (ITA) before withdrawal of an anti-thyroid drug (ATD) is useful for predicting relapse of Graves' disease (GD). We further investigated whether the ITA-PSV can be used for prediction of GD relapse after delivery in euthyroid women with GD who stopped ATD administration during mid- to late pregnancy. PATIENTS AND METHODS: ITA-PSV was monitored monthly for three months after delivery in 42 women with GD aged from 24 to 45 years old (mean+/-SE: 34.7+/-0.92 years old) who met the above criteria. To confirm the stability of the measurement, ITA-PSV was also measured monthly in 32 age-matched non-pregnant normal women and for three months after delivery in 10 age-matched women. RESULTS: ITA-PSV and thyroid volume were higher in women with GD immediately after delivery compared to normal women, but the levels of TSH receptor antibody (TRAb) and thyroid-stimulating antibody (TSAb) did not differ significantly between the two groups. Of the 42 patients, 23 had relapse of GD and the smoker/non-smoker ratio and thyroid volume in these patients immediately after delivery were significantly higher than those in the 19 patients who did not undergo relapse (10/23 vs. 0/19, p<0.0001; 24280.3+/-2280.9 vs. 19670.0+/ 2103.7mm(3), p=0.046), while ITA-PSV, TRAb and TSAb did not differ between the two groups of patients. The ITA-PSV ratio was calculated by dividing each value in the follow-up period by that obtained immediately after delivery. A significant increase in the mean ITA-PSV ratio occurred at least one month before the time of relapse (1.00+/-0.00 at -3 months before relapse vs. 1.46+/-0.12 at 1 month, p=0.010; 1.00+/-0.00 at -3 months vs. 1.77+/-0.13 at the time of relapse, p=0.0048). In contrast, there were no significant changes in this ratio during the follow-up period in non-relapse patients. CONCLUSION: Monthly measurement of ITA-PSV after delivery in remitted euthyroid women with GD may assist in early prediction of GD relapse. PMID- 20006463 TI - Preparation and pharmaceutical evaluation of new sustained-release capsule including starch-sponge matrix (SSM). AB - The focus of current study was to demonstrate a new sustained-release capsule including starch-sponge matrix (SSM) and to investigate how the pharmaceutical properties of SSM affect the drug release or its pharmacokinetic properties. Three representative drugs (uranine [UN], indomethacin [IMC] and nifedipine [NFP]) with different physicochemical properties (LogP(ow): 0.10, 1.18 and 3.23, respectively) were selected as model drugs. Model drug was dispersioned in pastelike cornstarch (starch glue) after heating 2.0-3.0% cornstarch suspension with electromagnetic wave at 2450 MHz (700 W) for l min. Then the drug mixture was encapsulated into a gratin capsule by a syringe, and the SSM including drug was prepared by means of a freeze-dried method. Essentially, drug-free SSM has a porous and netlike structure, and the distribution aspect of model drugs in the SSM depends on physicochemical properties between cornstarch glue and drugs. UN with much lower lipophilicity exists in continues phase of SSM, and IMC or NFP with a moderate or a higher lipophilicity exist in continues phase or porous space of the SSM. In the in vitro dissolution study, the release rate of drug from the SSM was mainly dependent on the lipophilicities of drugs, showing a rank order of the release rate of UN>IMC>NFP. In addition, the in vitro release rate for each drug was well regulated by changing the initial concentration of cornstarch suspension. In vivo absorption studies after intraduodenal administration of SSM capsule including model drug revealed that the sustained release effects also could be regulated by the initial concentration of starch suspension. Moreover, the sustained-release effect of SSM capsule was enhanced with an increase in the lipophilicity of drug, and local-residential and mucoadhesive properties of SSM in the intestine provided stable supply of drugs from the SSM. The SSM capsule we developed here shows promising results as an oral drug delivery system for sustained-release regulation or target specificity. PMID- 20006464 TI - The influence of fasudil on the epithelial-mesenchymal transdifferentiation of renal tubular epithelial cells from diabetic rats. AB - To investigate the influence of fasudil on the epithelial-mesenchymal transdifferentiation of renal tubular epithelial cells from diabetic rats and explore the mechanisms of this effect. Wistar rats were randomly divided into the following three groups: control, diabetes and fasudil-treatment. All rats were sacrificed after three months of feeding with or without fasudil treatment. Pathological changes to the glomeruli and renal interstitium were studied using Periodic acid-Schiff's staining and Masson staining, respectively. Expression of ROCK1, alpha-SMA, E-cadherin and the distribution of beta-catenin in rat renal cortex were revealed by immunohistochemistry. Changes in the MYPT1 phosphorylation profile and alpha-SMA, E-cadherin and membrane beta-catenin expression were revealed by western blot. Changes in the levels of ROCK1, E cadherin and total beta-catenin mRNA expression were analyzed by real-time PCR. Fasudil treatment notably attenuates renal interstitial fibrosis in diabetic rats. Compared to the control rats, diabetic rats showed elevated phosphorylation of MYPT1, increased expression of ROCK1 and alpha-SMA, decreased expression of E cadherin and membrane beta-catenin, and increased expression of ROCK1 and total beta-catenin mRNA, decreased expression of E-cadherin mRNA. Fasudil treatment of diabetic rats resulted in attenuated MYPT1 phosphorylation, decreased ROCK1 and alpha-SMA expression, increased E-cadherin and membrane beta-catenin expression, and reduced ROCK1 and total beta-catenin mRNA expression, increased expression of E-cadherin mRNA. In conclusion, fasudil may reduce the epithelial-mesenchymal transdifferentiation and renal interstitial fibrosis in diabetic rats through a mechanism by which ROCK activity is inhibited, which further facilitates the recovery of the cell-cell adhesions among renal tubular epithelial cells and adhesion complex formation. PMID- 20006466 TI - [The universities evaluation]. PMID- 20006465 TI - Histological substrate of human atrial fibrillation. AB - Histologic and ultrastructural examination of atrial tissue regarding the main entities responsible of human atrial fibrillation, is reported. The pathologic changes deriving from various disorders, like degenerative, inflammatory, ischemic diseases as well as from cardiac aging and hormonal imbalance are analysed. Structural changes associated with lone atrial fibrillation and investigated by atrial biopsy are also described, as being able to provide useful information on the disease's etiology, prognosis and treatment. PMID- 20006467 TI - Novel antimicrobial lipopeptides with long in vivo half-lives. AB - The pharmacokinetic (PK) properties of novel lipopeptides (semi-synthetic amphomycin analogues) with potent activity against Gram-positive organisms were evaluated in mice and rats following single intravenous (i.v.) and oral administration. Following oral administration at 50mg/kg, plasma concentrations of amphomycin analogues were <0.3-0.9microg/mL, suggesting that oral availability was low. Following i.v. administration (5-10mg/kg), the majority of lipopeptides demonstrated a long half-life (5.2-8.0h in mice and 4.6-7.1h in rats), low clearance (0.005-0.016mL/min in mice and 0.050-0.084mL/min in rats) and a volume of distribution indicative of extracellular penetration (0.118-0.339L/kg in mice and 0.121-0.133L/kg in rats). The area under the plasma concentration-time curve extrapolated to infinity (AUC(0-infinity)) for a 10mg/kg i.v. dose was determined to be 601.7-791.7microgh/mL in mice and 511.1-850.2microgh/mL in rats. The long half-life and low clearance observed with these novel lipopeptides indicate that drug serum concentrations will remain above the target minimal inhibitory concentration (MIC) levels for significant periods of time. When combined with the potent efficacy of these agents against Gram-positive organisms, the results of the present study support further development of these lipopeptide analogues towards clinical evaluation. PMID- 20006468 TI - Antibacterial and lipopolysaccharide (LPS)-neutralising activity of human cationic antimicrobial peptides against periodontopathogens. AB - In this study, we investigated the antibacterial activity of eight antimicrobial peptides (AMPs), comprising four human beta-defensins (HBDs), three human neutrophil defensins (HNPs) and the cathelicidin LL-37, against two representative periodontopathogens, Prevotella intermedia and Tannerella forsythia. The neutralising effect of these AMPs on expression of interleukin (IL)-1beta, IL-8 and intercellular adhesion molecule 1 (ICAM-1) induced by lipopolysaccharide (LPS) from P. intermedia and T. forsythia was also tested in THP-1 cells and human gingival fibroblasts. Prevotella intermedia was susceptible to HBD-3 and LL-37 but was resistant to HBD-1, HBD-2, HBD-4, HNP-1, HNP-2 and HNP 3 at concentrations up to 10microM. However, all of the AMPs except HNP-2 at 5microM significantly inhibited the expression of IL-1beta, IL-8 and ICAM-1 induced by P. intermedia LPS. Tannerella forsythia showed marked susceptibility to the AMPs tested in the following order: LL-37, HBD-3, HBD-2, HBD-1, HNP-1 and HBD-4. All of the AMPs except HNP-3 had significant neutralising effects on T. forsythia LPS activity. The AMPs showing LPS-neutralising activity inhibited LPS binding to the cells. These results suggest that AMPs may be considered as preventive and therapeutic agents against mixed bacterial infections such as periodontitis by eliminating the pathogens themselves as well as reducing the activity of LPS. PMID- 20006469 TI - Bactericidal activity of daptomycin versus vancomycin in the presence of human albumin against vancomycin-susceptible but tolerant methicillin-resistant Staphylococcus aureus (MRSA) with daptomycin minimum inhibitory concentrations of 1-2microg/mL. AB - This study explored the influence of vancomycin tolerance and protein binding on the bactericidal activity of vancomycin versus daptomycin (protein binding 36.9% vs. 91.7%, respectively) against four vancomycin-tolerant methicillin-resistant Staphylococcus aureus (MRSA) [minimum inhibitory concentration/minimum bactericidal concentration (MIC/MBC)=0.5/16, 1/32, 2/32 and 1/32microg/mL for vancomycin and 1/1, 1/2, 2/2 and 2/4microg/mL for daptomycin]. Killing curves were performed with vancomycin/daptomycin concentrations equal to serum peak concentrations (C(max)) (65.70/98.60microg/mL) and trough concentrations (C(min)) (7.90/9.13microg/mL) in the presence and absence of a physiological human albumin concentration (4g/dL), controlled with curves with the theoretical free drug fraction of vancomycin/daptomycin C(max) (41.45/8.18microg/mL) and C(min) (4.98/0.76microg/mL). Vancomycin C(max) and C(min) concentrations, regardless of the media, showed a bacteriostatic profile not reaching a reduction of 99% or 99.9% of the initial inocula during the 24-h experimental time period. Daptomycin antibacterial profiles significantly differed when testing C(max) and C(min). C(max) was rapidly bactericidal (< or =4h) with >5 log(10) reduction in the initial inocula for all strains, regardless of the presence or not of albumin or the use of concentrations similar to free C(max). C(min) exhibited similar final colony counts at 0h and 24h in curves with albumin, but with >3 log colony forming units (CFU)/mL reduction at < or =4h for strains with an MIC of 1microg/mL and ca. 2 logCFU/mL reduction at < or =6h for strains with an MIC of 2microg/mL. This activity was significantly higher than the activity of the free C(min) fraction. The results of this study reinforce the idea that pharmacodynamics using concentrations calculated using reported protein binding are unreliable. Daptomycin exhibited rapid antibacterial activity against vancomycin-tolerant MRSA isolates even against those with high daptomycin MICs in the presence of physiological albumin concentrations. PMID- 20006470 TI - In vitro drug susceptibility of 2275 clinical non-tuberculous Mycobacterium isolates of 49 species in The Netherlands. AB - In this study, 2275 clinical isolates of 49 species of non-tuberculous mycobacteria isolated in The Netherlands were subjected to standardised drug susceptibility testing using the Middlebrook 7H10 agar dilution method. Clarithromycin and rifabutin were most active, with 87% and 83% of all isolates, respectively, being susceptible. Susceptibility to ciprofloxacin (44%) and amikacin (32%) was limited and was mostly restricted to Mycobacterium kansasii, Mycobacterium xenopi, Mycobacterium fortuitum and phylogenetically related species. Susceptibility to isoniazid (0.5%), rifampicin (37%), ethambutol (35%) and streptomycin (33%) was rare; susceptibility to cycloserine, clofazimine and prothionamide was generally restricted to slow growers, although prothionamide also had activity against M. fortuitum and related species. Significant discrepancies between in vitro and in vivo activity exist. To improve the utility of drug susceptibility testing, the selection of drugs should be changed to more drugs with proven clinical efficacy correlating with in vitro susceptibility. PMID- 20006471 TI - Colistin therapy for microbiologically documented multidrug-resistant Gram negative bacterial infections: a retrospective cohort study of 258 patients. AB - It is unclear whether the effectiveness of polymyxins depends on the site of infection, the responsible pathogen, dosage, and monotherapy vs. combination therapy. We investigated colistin therapy in a large, retrospective, single centre, cohort study. Primary analysis outcomes were infection outcome, survival and nephrotoxicity. Over a 7-year period (October 2000 to October 2007), 258 patients received intravenous (i.v.) colistin for at least 72h for microbiologically documented multidrug-resistant Gram-negative bacterial infections, comprising 170 (65.9%) Acinetobacter baumannii, 68 (26.4%) Pseudomonas aeruginosa, 18 (7.0%) Klebsiella pneumoniae, 1 (0.4%) Stenotrophomonas maltophilia and 1 (0.4%) Enterobacter cloacae. Cure of infection occurred in 79.1% of patients, nephrotoxicity in 10% and hospital survival in 65.1%. In the multivariate analysis, independent predictors of survival were colistin average daily dose [adjusted odds ratio (aOR)=1.22, 95% confidence interval (CI) 1.05-1.42] and cure of infection (aOR=9, 95% CI 3.6-23.1), whilst the proportion of creatinine change (aOR=0.21, 95% CI 0.1-0.45), Acute Physiology and Chronic Health Evaluation (APACHE) II score (aOR=0.89, 95% CI 0.84-0.95) and haematological disease (aOR=0.23, 95% CI 0.08-0.66) were associated with mortality. Effectiveness of colistin was not dependent on the type of pathogen. No independent predictors for nephrotoxicity were observed. The findings of the largest cohort study to date on i.v. colistin show that colistin is a valuable antibiotic with acceptable nephrotoxicity and considerable effectiveness that depends on the daily dosage and infection site. PMID- 20006472 TI - High-level ciprofloxacin resistance among hospital-adapted Enterococcus faecium (CC17). AB - Hospital-adapted Enterococcus faecium differ from their colonising variants in humans and animals by additional genomic content. Molecular typing based on multilocus sequence typing (MLST) allows allocation of isolates to specific clonal complexes (CCs), such as CC17 for hospital-adapted strains. Acquired ampicillin resistance is a specific feature of these hospital isolates, especially in Europe. A few recent reports have described acquired high-level ciprofloxacin resistance as a supposed feature of hospital-adapted E. faecium strains. In the present retrospective analysis, ciprofloxacin minimum inhibitory concentrations (MICs) of 609 clinical isolates from German hospital patients (1997-2007) were determined and a breakpoint for high-level resistance was deduced (>16mg/L). Acquired high-level ciprofloxacin resistance was distributed among isolates of 26 different MLST types (all CC17), indicating a wide prevalence of this acquired resistance trait among the hospital-adapted E. faecium population. High-level ciprofloxacin resistance was linked to gyrA and parC mutations in 98 investigated isolates. Eleven different allele types or combinations thereof were identified. Their allocation to specific MLST and pulsed-field gel electrophoresis (PFGE) types revealed differences in the emergence and spread of corresponding mutations and strains. PMID- 20006474 TI - EEG source analysis of chronic fatigue syndrome. AB - Sixty-one dextral, unmedicated women with chronic fatigue syndrome (CFS) diagnosed according to the Fukuda criteria (1994) and referred for investigation by rheumatologists and internists were studied with quantitative EEG (43 channels) at rest with eyes open and during verbal and spatial cognitive activation. The EEGs from the patients were compared with recordings from 80 dextral healthy female controls. Only those subjects who could provide 20 1-s artefact-free segments of EEG were admitted into the study. The analysis consisted of the identification of the spatial patterns in the EEGs that maximally differentiated the two groups and the estimation of the cortical source distributions underlying these patterns. Spatial patterns were analyzed in the alpha (8-13Hz) and beta (14-20Hz) bands and the source distributions were estimated using the Borgiotti-Kaplan BEAMFORMER algorithm. The results indicate that the spatial patterns identified were effective in separating the two groups, providing a minimum correct retrospective classification rate of 72% in both frequency bands while the subjects were at rest to a maximum of 83% in the alpha band during the verbal cognitive condition. Underlying cortical source distributions showed significant differences between the two groups in both frequency bands and in all cognitive conditions. Lateralized cortical differences were evident between the two groups in the both frequency bands during both the verbal and spatial cognitive conditions. During these active cognitive conditions, the CFS group showed significantly greater source-current activity than the controls in the left frontal-temporal-parietal regions of the cortex. PMID- 20006473 TI - Old class but new dimethoxy analogue of benzimidazole: a bacterial topoisomerase I inhibitor. AB - New antimicrobials are needed to combat drug resistance and have often been equated with the identification and exploitation of novel targets. This study focused on the synthesis of new benzimidazole analogues with improved DNA minor groove-binding affinity and having lower cytotoxicity to mammalian cells as well as selective targeting of bacterial DNA over host DNA. 5-(4-Methylpiperazin-1-yl) 2-[2'-(3,4-dimethoxyphenyl)-5'-benzimidazolyl]benzimidazole (DMA) cleared bacterial infections from mammalian cell culture without apparent cytotoxicity to mammalian cells. Moreover, DMA inhibited microbial topoisomerase over mammalian topoisomerase, with a 50% inhibitory concentration (IC50) value for human topoisomerase I of >54microM compared with an IC50 of <10microM for Escherichia coli topoisomerase I in vitro. PMID- 20006475 TI - VEGF-mediated signal transduction in lymphatic endothelial cells. AB - The VEGF family of angiogenic ligands consists of VEGFA, VEGFB, VEGFC, VEGFD and placenta growth factor, PlGF. These growth factors bind in an overlapping pattern to three receptor tyrosine kinases, denoted VEGFR1, VEGFR2 and VEGFR3. Originally, VEGFA (the prototype VEGF) was described as a master regulator of vascular endothelial cell biology in vitro and in vivo, transducing its effect through VEGFR2. VEGFA, VEGFB and PlGF bind to VEGFR1, which is a negative regulator of endothelial cell function at least during embryogenesis. VEGFC and VEGFD were identified as lymphatic endothelial factors, acting via VEGFR3. With time, the very clear distinction between the roles of the VEGF ligands in angiogenesis/lymphangiogenesis has given way for a more complex pattern. It seems that the biology of the different VEGFR2 and VEGFR3 ligands overlaps quite extensively and that both receptor types contribute to angiogenesis as well as lymphangiogenesis. This paradigm shift in our understanding is due to the access to more sophisticated reagents and techniques revealing dynamic and plastic expression of ligands and receptors in different physiological and pathological conditions. Moreover, knowledge on the important role of VEGF coreceptors, the neuropilins, in regulating the responsiveness to VEGF has changed our perception on the mechanism of VEGF signal transduction. This review will primarily focus on the properties of VEGR3, its signal transduction and the resulting biology. PMID- 20006476 TI - The fine structure of the developing oocyst of Pterospora floridiensis (Apicomplexa, Urosporidae). AB - Developing oocysts of the gregarine Pterospora floridiensis Landers 2001 were examined by transmission electron microscopy. Each oocyst had an outer capsule and an inner capsule that contained 8 sporozoites. In early stages of development the inner capsular wall was separated from the developing sporozoites and residual mass, and was not appressed to the sporozoites. Early stage sporozoites were connected to a residual mass and were filled with endoplasmic reticulum, golgi and numerous developing secretory vesicles. In late stages of oocyst and sporozoite development, the inner capsular wall was closely appressed to the sporozoite surface. The inner capsular wall was approximately 60-100 nm thick and the outer capsular wall was approximately 160-320 nm thick. There were no extensions on the outer wall for which the genus was named. Late stage sporozoites had no residual mass connection, were more electron dense, and contained three distinct types of dense secretory structures: 1) small oval/spherical dense vesicles, 2) large (350-400 nm) vesicles near the anterior end, and 3) elongated dense tubular bodies that converged at the apex. Few ultrastructural reports exist of developing gregarine oocysts and sporozoites, and as more studies are completed these morphological characteristics may be important in interpreting molecular phylogenetic analyses. PMID- 20006477 TI - Semantic SenseLab: Implementing the vision of the Semantic Web in neuroscience. AB - OBJECTIVE: Integrative neuroscience research needs a scalable informatics framework that enables semantic integration of diverse types of neuroscience data. This paper describes the use of the Web Ontology Language (OWL) and other Semantic Web technologies for the representation and integration of molecular level data provided by several of SenseLab suite of neuroscience databases. METHODS: Based on the original database structure, we semi-automatically translated the databases into OWL ontologies with manual addition of semantic enrichment. The SenseLab ontologies are extensively linked to other biomedical Semantic Web resources, including the Subcellular Anatomy Ontology, Brain Architecture Management System, the Gene Ontology, BIRNLex and UniProt. The SenseLab ontologies have also been mapped to the Basic Formal Ontology and Relation Ontology, which helps ease interoperability with many other existing and future biomedical ontologies for the Semantic Web. In addition, approaches to representing contradictory research statements are described. The SenseLab ontologies are designed for use on the Semantic Web that enables their integration into a growing collection of biomedical information resources. CONCLUSION: We demonstrate that our approach can yield significant potential benefits and that the Semantic Web is rapidly becoming mature enough to realize its anticipated promises. The ontologies are available online at http://neuroweb.med.yale.edu/senselab/. PMID- 20006479 TI - Enhancement of aerobic degradation of benzoate and 2-chlorobenzoate by adapted activated sludge. AB - Activated sludge from the Stickney Water Reclamation Plant of the Metropolitan Water Reclamation District of Greater Chicago was adapted in the laboratory to either benzoate or 2-chlorobenzoate as the sole carbon source in sequencing batch reactors with a 48-h feed-aerate-settle-draw cycle and a mean cell residence time (MCRT) of 10 days. Benzoate degradation increased by more than 80-fold after 7 MCRTs compared to unadapted activated sludge. A greater than 15-fold increase in 2-chlorobenzoate metabolism occurred after adaptation for about 5-7 MCRTs. For each substrate the maximum rate measured for adapted cultures was near or above the highest previously reported in the literature. For both adapted and unadapted sludges, benzoate metabolism was considerably faster than that of 2 chlorobenzoate, and for both substrates the rate of metabolism increased incrementally with time of adaptation. As expected, addition of the benzoate adapted sludge to unadapted sludge enhanced the latter's ability to degrade benzoate. PMID- 20006478 TI - Minimal influence of water and nutrient content on the bacterial community composition of a maritime Antarctic soil. AB - Bacterial community composition was determined by culture-independent PCR-based methods in two soils differing markedly in their water, C, N and P contents sampled from Mars Oasis on Alexander Island, western Antarctic Peninsula. 16S rRNA sequences of the phyla Actinobacteria, Cyanobacteria, alpha-Proteobacteria and Acidobacteria were commonly (> 8% frequency) obtained from soil. Those of beta-, gamma- and delta-Proteobacteria, Chloroflexi, Bacteroidetes, Verrucomicrobia, Planctomycetes, Gemmatimonadetes and Firmicutes were less frequent. Comparisons of slopes of collector's curves and the Shannon-Weiner diversity index indicated no difference in overall bacterial diversity between the two soils, although sequences of delta-Proteobacteria and the cyanobacterial genus Leptolyngbya were more commonly derived from the soil with the higher water and nutrient content. The data suggest that different levels of soil water, C, N and P have only a minor effect on the bacterial community composition of maritime Antarctic soils. PMID- 20006480 TI - Identification, cloning, expression, and purification of Francisella lpp3: an immunogenic lipoprotein. AB - The severe and fatal human disease, tularemia, results from infection with the Gram-negative pathogen Francisella tularensis. Identification of surface outer membrane proteins, specifically lipoproteins, has been of interest for vaccine development and understanding the initiation of disease. We sought to identify Francisella live vaccine strain lipoproteins that could be a component of a subunit vaccine and have adjuvant properties as TLR2 agonists. We have identified a membrane lipoprotein of Francisella LVS isolated by sarkosyl extraction and gel filtration chromatography that is recognized by sera from LVS-vaccinated individuals and tularemia patients, indicating its potential diagnostic value. Sequencing of the protein by mass spectrometry indicated that it encodes the FTL_0645 open reading frame of F. holarctica LVS, which is 100% identical/homologous to FTT1416c of F. tularensis Schu S4. The predicted 137 amino acid lipoprotein encoded by FTL_0645 ORF, was expressed in Escherichia coli, purified, and demonstrated to be a lipoprotein. This recombinant lipoprotein, named Flpp3, was able to activate TLR2 and induce an immunogenic response in mice, suggesting that the E. coli-expressed Flpp3 is palmitoylated and closely resembles the native protein in structure and immunogenicity. Taken together, these data suggest that Flpp3 could be a candidate for inclusion in a F. tularensis vaccine. PMID- 20006482 TI - Reducing irreducible complexity: divergence of quaternary structure and function in macromolecular assemblies. AB - The bacterial flagellar system is an intricate assembly (containing approximately 40 different proteins) that is involved in both protein secretion and bacterial motility. It has also become the icon of the neo-creationist movement in the United States, with the argument that it shows 'irreducible complexity' and could not have been the product of evolution. Recent studies provide new insights into the evolution of the flagellar system and lead to the suggestion that the divergence of quaternary structure in protein assemblies may be an underappreciated mechanism for rapid evolutionary divergence. Work on the enzyme FucU, involved in fucose metabolism, may suggest similar conclusions. PMID- 20006481 TI - TOR-dependent control of autophagy: biting the hand that feeds. AB - Induction of autophagy in response to starvation is a highly conserved ability of eukaryotic cells, indicating a crucial and ancient role of this process in adapting to nutrient conditions. The target of rapamycin (TOR) pathway is major conduit for such signals, and in most cell types TOR activity is necessary and sufficient to suppress autophagy under favorable growth conditions. Recent studies have begun to reveal how TOR activity is regulated in response to nutritional cues, and are shedding new light on the mechanisms by which TOR controls the autophagic machinery. In addition, a variety of signals, stressors and pharmacological agents that induce autophagy independent of nutrient conditions have been identified. In some cases these signals appear to have been spliced into the core TOR pathway, whereas others are able to bypass the control mechanisms regulated by TOR. Increasing evidence is pointing to an important role for both positive and negative feedback loops in controlling this pathway, leading to an emerging view that TOR signaling not only regulates autophagy but is also highly sensitive to cellular rates of autophagy and other TOR-dependent processes. PMID- 20006483 TI - Islam and harm reduction. AB - Although drugs are haram and therefore prohibited in Islam, illicit drug use is widespread in many Islamic countries throughout the world. In the last several years increased prevalence of this problem has been observed in many of these countries which has in turn led to increasing injecting drug use driven HIV/AIDS epidemic across the Islamic world. Whilst some countries have recently responded to the threat through the implementation of harm reduction programmes, many others have been slow to respond. In Islam, The Quran and the Prophetic traditions or the Sunnah are the central sources of references for the laws and principles that guide the Muslims' way of life and by which policies and guidelines for responses including that of contemporary social and health problems can be derived. The preservation and protection of the dignity of man, and steering mankind away from harm and destruction are central to the teachings of Islam. When viewed through the Islamic principles of the preservation and protection of the faith, life, intellect, progeny and wealth, harm reduction programmes are permissible and in fact provide a practical solution to a problem that could result in far greater damage to the society at large if left unaddressed. PMID- 20006484 TI - One-pot construction of mediatorless bi-enzymatic glucose biosensor based on organic-inorganic hybrid. AB - A new methodology for the fabrication of bienzymatic amperometric glucose biosensor based on the use of an organic-inorganic hybrid is presented. The fabrication involves a self-assembly directed one-pot electrochemical process. Bi enzymes, horseradish peroxidase (HRP) and glucose oxidase (GOx) are immobilized into the porous and electroactive silica-polyaniline hybrid composite through electrochemical polymerization of N[3-(trimethoxysilyl)propyl]aniline in the presence of enzymes. The modified electrode is designated as PTMSPA/HRP-GOx. The direct electron transfer of HRP is achieved at the modified electrode. Also, the electrode exhibits excellent bio-electro-catalytic activity for the reduction of hydrogen peroxide. The response current at PTMSPA/HRP-GOx modified electrode revealed a good linear relationship with concentration of glucose range between 1 and 20mM with a response time of 7s. Thus, the modified electrode shows the combined advantages of polyaniline and silica networks through synergistic influence from the individual components. The PTMSPA assembly has shown the potential for a third generation amperometric biosensor. PMID- 20006485 TI - A biofuel cell with a single-walled carbon nanohorn-based bioanode operating at physiological condition. AB - Single-walled carbon nanohorns (SWNHs), a new type of carbon nanomaterials, possess excellent catalytic properties, high-purity, and low toxicities, which make them suitable for bioelectrochemical application. Here a biofuel cell anode has been developed by using SWNHs as the support for redox mediator and biocatalyst for the first time. Cyclic voltammetric results show SWNHs promotes the electropolymerization of methylene blue (MB) and the resulted nanocomposite (poly MB-SWNHs) exhibits prominent catalytic ability to the oxidation of nicotinamide adenine dinucleotide. Glucose dehydrogenase was then immobilized on the poly MB-SWNHs modified electrode for the oxidation of glucose. Employing Pt nanoparticles supported on functionalized TiO(2) colloidal spheres with nanoporous surface as cathode catalyst, the as-assembled glucose/O(2) biofuel cell operate at the physiological condition with good performance. PMID- 20006486 TI - Targeting the PI3K signaling pathway in cancer. AB - The phosphoinositide 3-kinase (PI3K) pathway is activated in a variety of different human cancers, and inhibitors of this pathway are under active development as anti-cancer therapeutics. In this review, we discuss the data supporting the use of PI3K pathway inhibitors in genetically and clinically defined cancers. This review focuses on their efficacy as single agents and in combination with other targeted therapies, specifically those targeting the MEK ERK signaling pathway. PMID- 20006487 TI - Multimodal approach using oncolytic adenovirus, cetuximab, chemotherapy and radiotherapy in HNSCC low passage tumour cell cultures. AB - Head and neck squamous cell carcinoma (HNSCC) is a common and often devastating disease without curative treatment when advanced or recurrent. The aim of this study was to assess whether capsid-modified oncolytic adenoviruses have therapeutic efficacy in HNSCC low passage tumour cell cultures and if it could be further improved by combination with cetuximab, radiotherapy and chemotherapy. We investigated which adenoviral capsid modifications allow best gene transfer and cell killing of HNSCC substrates. Gene transfer was assessed using replication deficient adenoviruses expressing luciferase. Cell killing was studied in vitro and in vivo using oncolytic adenoviruses, which kill tumour cell by viral replication. The most effective capsid-modified oncolytic adenoviruses were combined with HNSCC standard therapies and their efficacy was assessed in vitro as well as in vivo. Cell killing was assessed in vitro by MTS assay and in vivo by HNSCC subcutaneous tumour growth follow-up in nude mice. Cetuximab treatment was found to enrich CD133+ and CD44+ tumour-initiating type cells in tumours grown in mice. Capsid-modified viruses showed increased transduction and oncolysis of HNSCC substrates in comparison to Ad5-based agents. Polylysine (pK7) modified oncolytic virus resulted in significant tumour growth reduction in vivo. Combination of chemotherapy (cisplatin and 5-fluorouracil), radiotherapy and cetuximab with oncolytic adenovirus therapy resulted in further increases in cell killing effect in vitro and complete eradication of tumours in vivo. Our pre clinical data suggest that it is feasible and efficacious to combine oncolytic adenoviruses with HNSCC standard therapies into a multimodality treatment regimen for clinical testing in HNSCC patients. PMID- 20006488 TI - Pain assessment in the elderly. AB - Admissions to hospital for patients aged over 65 years are three times higher than for younger patients for all medical and surgical wards. Older people are often excluded from trials on pain assessment and treatment because of cognitive or sensory impairments. Professionals tend to underestimate pain needs, under prescribe and under-medicate in general and in older people in particular. Where studies have included older people, the benefit of treatment is similar regardless of age. The first step in managing acute pain is through its assessment. Although pain is a subjective experience, pain rating scales are valid and reliable when used appropriately. Older people demonstrate some differences in reporting pain that may be attributable to a range of factors including biology, culture, religion, ethnicity, cognitive impairment, organisation or social context. Attitudinal barriers are also relevant because these include a persistent belief that older people experience less pain than other age groups. Not surprisingly, older people themselves might believe that pain is something to be endured, strong analgesics lead to addiction, complaining about pain is a sign of personal weakness and pain is an inevitable part of aging. Undertreatment of pain can lead to the development of chronic pain syndromes that can prove difficult to treat and adversely affect long-term quality of life. Effective treatment is paramount because of the increased morbidity and mortality associated with undertreated pain. PMID- 20006489 TI - Assessment and treatment of elderly patients with cancer. AB - In the US, the total projected cancer incidence will increase by 45% yearly from 2010 to 2030. The majority of these neoplasms will affect the elderly. The National Institute on Aging has characterised the aging of our society as a "silver tsunami". Thus, the management of cancer in the older aged person represents a priority for health care delivery in the immediate future. Aging, age-related changes, frailty, prolongation of active life expectancy, prevention of Adverse Drugs Reactions (ADRs), Comprehensive Geriatric Assessment (CGA), and individualised treatment in the future will become the most widely used concepts in the management of cancer patients. PMID- 20006490 TI - On energy consumption for size-reduction and yields from subsequent enzymatic saccharification of pretreated lodgepole pine. AB - This study investigated the effects of chemical pretreatment and disk-milling conditions on energy consumption for size-reduction and the efficiency of enzymatic cellulose saccharification of a softwood. Lodgepole pine wood chips produced from thinnings of a 100-year-old unmanaged forest were pretreated by hot water, dilute-acid, and two SPORL processes (Sulfite Pretreatment to Overcome Recalcitrance of Lignocellulose) at acid charge on oven dry (od) wood of 0% and 2.21%. The pretreated wood chips were then milled using a laboratory disk mill under various solids-loadings and disk-plate gaps to produce substrates for enzymatic hydrolysis. We found that post-chemical-pretreatment size-reduction of forest biomass can decrease size-reduction energy consumption by 20-80% depending on the pretreatment applied under 20% solids-loading and a disk-plate gap of 0.76 mm in milling. SPORL with a sodium bisulfite charge of 8% and sulfuric acid charge of 2.21% on wood was the most effective in decreasing size-reduction energy consumption. Solids-loading had the most significant effect on disk milling energy. When solids-loading was reduced from 30% to 3%, disk-milling energy could be decreased by more than a factor of 10 for wood chips pretreated by both SPORL and dilute-acid at an acid charge of 2.21%. The enzymatic hydrolysis glucose yields (EHGY) from the substrates produced by all pretreatments were independent of the solids-loading in milling, indicating that these energy savings in size-reduction can be realized without affecting EHGY. When wood chips were pretreated by SPORL with 2.21% acid charge, size-reduction energy consumption was decreased to less than 50 Wh/kg od wood at a practical solids-loading of approximately 10-20%, equivalent to that used in size-reduction of agriculture biomass, with excellent EHGY of about 370 g per kg od wood. Similar effects on size-reduction energy savings and excellent EHGY were also achieved when large disk-plate gaps (up to 1.52 mm studied) were applied in disk milling of wood chips pretreated by SPORL with acid. PMID- 20006491 TI - Bio-conversion rate and optimum harvest intervals for Moina australiensis using digested piggery effluent and Chlorella vulgaris as a food source. AB - The bio-conversion rate of Moina australiensis fed with Chlorella vulgaris grown on digested piggery effluent at three different feeding rates was determined and a 2, 3 and 4-day harvest interval strategy was investigated. This study indicates that C. vulgaris is a suitable food source for M. australiensis. A significant difference (P < or = 0.001) in the feeding rate against mean total populations was found among treatments. The increase in the amount of algae fed accelerated the production rate, and the population density peaked faster in the high C. vulgaris fed treatment. The BCR calculated from this experiment indicates that for every 1000 mg of C. vulgaris fed there was an increase of 437.9 mg of M. australiensis biomass produced. A significant difference (P < or = 0.001) in biomass production among the different harvest interval treatments was observed. The 2-day harvest interval treatment produced 7.78 g of M. australiensis followed by 6.89 g in the 3 day and 5.01 g in the 4-day harvest interval treatment. This study provides strong evidence that M. australiensis can utilise the bacterial blooms and bio-films associated with digested piggery effluent as a food source. PMID- 20006492 TI - Treatment of waste gas containing low concentration of dimethyl sulphide (DMS) in a bench-scale biofilter. AB - Biological treatment of dimethyl sulphide (DMS) was investigated in a bench-scale biofilter, packed with compost along with wood chips, and enriched with DMS degrading microorganism Bacillus sphaericus. The biofilter could remove 62-74% of the inlet DMS, at an optimum loading of 0.484 g/m(3)/h with optimum empty bed contact time (EBCT) of 384 s and an average moisture range of 65-70%. The biodegradative products of DMS were sulphide, thiosulphate and sulphate. Evaluation of microbiological status of the biofilter indicated the presence of other bacterial cultures viz. Paenibacillus polymyxa, and Bacillus megaterium, besides B. sphaericus. PMID- 20006493 TI - Pretreatment and enzymatic hydrolysis of recovered fibre for ethanol production. AB - Recovered fibre from pulp mills represents a potentially significant feedstock for conversion to ethanol. Enzymatic hydrolysis of untreated recovered fibre (86.5 Kappa, 13% lignin) resulted in a hexose yield of approximately 23%, which highlighted the need for an effective pretreatment. Recovered fibre was pretreated as a substrate for enzymatic hydrolysis using oxygen delignification. An experimental design was used to optimize temperature (90-150 degrees C), caustic loading (2-10%), and reaction time (20-60 min). The post-delignification Kappa values ranged from 76.7 (11.5% lignin) under the mildest pretreatment conditions, to 20 (3% lignin) under the most severe pretreatment conditions. The effect of caustic load appears to have an increased effect at higher temperatures, with the Kappa numbers ranging from 76.7 (90 degrees C, 2% caustic, 20 min) to 56.0 (150 degrees C, 2% caustic, 20 min) and from 64.7 (90 degrees C, 10% caustic, 20 min) to 38.0 (150 degrees C, 10% caustic, 60 min). These changes in Kappa number reflect changes in the lignin fraction of 3.1% and 4%, respectively. Increasing the caustic load from 2% to 10% decreased the oxygen delignification yield from 93.5% to 87.9% at 90 degrees C and 20 min reaction time, and 80.3% to 74.7% at 150 degrees C. The effect of time on oxygen delignification yield was found to be most significant in the first twenty minutes, which correlates with the drop in Kappa number that was observed. The pretreated fibre was subjected to enzymatic hydrolysis using commercial enzymes Celluclast (80FPU/mL, 20.1CBU/mL) and Novozym (640.5 CBU/mL). A series of enzyme loadings ranging from 19 to 77 FPU/g were utilized on solids loading ranging from 20 to 100g (dry fibre)/L. Based on the pretreatment and hydrolysis results an empirical model was developed that can predict hydrolysis sugar concentrations based on the Kappa number, enzyme loading, and initial recovered fibre concentration. PMID- 20006494 TI - Bioethanol from lignocellulosics: Status and perspectives in Canada. AB - Canada has invested significantly in the development of a domestic bioethanol industry, and it is expected that bioethanol from lignocellulosics will become more desirable to the industry as it expands. Development of the Canadian industry to date is described in this paper, as are examples of domestic research programs focused on both bioconversion and thermochemical conversion to generate biofuels from lignocellulosic biomass. The availability of lignocellulosic residues from agricultural and forestry operations, and the potential biofuel production associated with these residues, is described. The policy tools used to develop the domestic bioethanol industry are explored. A residue-based process could greatly extend the potential of the bioethanol industry in Canada. It is estimated that bioethanol production from residual lignocellulosic feedstocks could provide up to 50% of Canada's 2006 transportation fuel demand, given ideal conversion and full access to these feedstocks. Utilizing lignocellulosic biomass will extend the geographic range of the bioethanol industry, and increase the stability and security of this sector by reducing the impact of localized disruptions in supply. Use of disturbance crops could add 9% to this figure, but not in a sustainable fashion. If pursued aggressively, energy crops ultimately could contribute bioethanol at a volume double that of Canada's gasoline consumption in 2006. This would move Canada towards greater transportation fuel independence and a larger role in the export of bioethanol to the global market. PMID- 20006495 TI - 1-(3-Aryloxyaryl)benzimidazole sulfones are liver X receptor agonists. AB - A series of 1-(3-aryloxyaryl)benzimidazoles incorporating a sulfone substituent (6) was prepared. High affinity LXR ligands were identified (LXRbeta binding IC(50) values <10nM), some with excellent agonist potency and efficacy in a functional assay of LXR activity measuring ABCA1 mRNA increases in human macrophage THP1 cells. The compounds were typically stable in liver microsome preparations and had good oral exposure in mice. PMID- 20006497 TI - 2,3,5-Trisubstituted pyridines as selective AKT inhibitors-Part I: Substitution at 2-position of the core pyridine for ROCK1 selectivity. AB - 2,3,5-Trisubstituted pyridines have been designed as potent AKT inhibitors that are selective against ROCK1 based on the comparison between AKT and ROCK1 structures. Substitution at the 2-position of the core pyridine is the key element to provide selectivity against ROCK1. An X-ray co-crystal structure of 9p in PKA supports the proposed rationale of ROCK1 selectivity. PMID- 20006496 TI - Microwave assisted synthesis and in vitro cytotoxicities of substituted (Z)-2 amino-5-(1-benzyl-1H-indol-3-yl)methylene-1-methyl-1H-imidazol-4(5H)-ones against human tumor cell lines. AB - The synthesis of several novel substituted (Z)-2-amino-5-(1-benzyl-1H-indol-3 yl)methylene-1-methyl-1H-imidazol-4(5H)-ones structurally related to aplysinopsin have been carried out under microwave irradiation and conventional heating methods. The analogs 3a, 3b, 3d-3g,3k and 3l were evaluated for their in vitro cytotoxic activity against an NCI 60 human tumor cell line panel. Compound 3f exhibited good growth inhibitory properties against all but four of the human cancer cell lines examined, and afforded LC(50) values <10microM for 30% of the cell lines in the panel. Compound 3e was an effective inhibitor of leukemia, CNS, melanoma, and breast cancer cell growth, but generally less effective as a cytotoxic agent. Thus, the aplysinopsin analog 3f was regarded as a useful lead compound for further structural optimization. PMID- 20006498 TI - Novel spiropiperidine-based stearoyl-CoA desaturase-1 inhibitors: Identification of 1'-{6-[5-(pyridin-3-ylmethyl)-1,3,4-oxadiazol-2-yl]pyridazin-3-yl}-5 (trifluoromethyl)-3,4-dihydrospiro[chromene-2,4'-piperidine]. AB - Cyclization of the benzoylpiperidine in lead compound 2 generated a series of novel and highly potent spiropiperidine-based stearoyl-CoA desaturase (SCD)-1 inhibitors. Among them, 1'-{6-[5-(pyridin-3-ylmethyl)-1,3,4-oxadiazol-2 yl]pyridazin-3-yl}-5-(trifluoromethyl)-3,4-dihydrospiro[chromene-2,4'-piperidine] (19) demonstrated the most powerful inhibitory activity against SCD-1, not only in vitro but also in vivo (C57BL/6J mice). With regard to the pharmacological evaluation, 19 showed powerful reduction of the desaturation index in the plasma of C57BL/6J mice on a non-fat diet after a 7-day oral administration (q.d.) without causing notable abnormalities in the eyes or skin up to the highest dose (3mg/kg) in our preliminary analysis. PMID- 20006499 TI - Structure and property based design of factor Xa inhibitors: pyrrolidin-2-ones with monoaryl P4 motifs. AB - Structure and property based drug design was exploited in the synthesis of sulfonamidopyrrolidin-2-one-based factor Xa inhibitors, incorporating neutral and basic monoaryl P4 groups, ultimately producing potent inhibitors with effective levels of anticoagulant activity and extended oral pharmacokinetic profiles. However, time dependant inhibition of Cytochrome P450 3A4 was a particular issue with this series. PMID- 20006500 TI - Tetrasubstituted pyridines as potent and selective AKT inhibitors: Reduced CYP450 and hERG inhibition of aminopyridines. AB - The synthesis and evaluation of tetrasubstituted aminopyridines, bearing novel azaindazole hinge binders, as potent AKT inhibitors are described. Compound 14c was identified as a potent AKT inhibitor that demonstrated reduced CYP450 inhibition and an improved developability profile compared to those of previously described trisubstituted pyridines. It also displayed dose-dependent inhibition of both phosphorylation of GSK3beta and tumor growth in a BT474 tumor xenograft model in mice. PMID- 20006501 TI - A profile of the in vitro anti-tumor activity of imidazolium-based ionic liquids. AB - The anti-cancer activity and cytotoxicity of imidazolium-based ionic liquids has been determined for the first time via NCI's in vitro 60 human tumor cell lines. The preliminary SAR showed that the chain length of alkyl substitution at N-3 position of imidazole ring plays crucial role towards anti-tumor activity and cytotoxicity of these ionic liquids. The ionic liquids with alkyl substitution of C-12 chain length were found to be effective against all 60 tumor cell lines and show very low cytotoxicity in most of the cases. Further increase in chain length resulted in enhanced growth inhibition of tumor cell lines as well as high cytotoxicity. Interestingly, active compounds 1-dodecyl-3-methylimidazolium chloride (8), 1-dodecyl-3-methylimidazolium tetrafluoroborate (9), 1-hexadecyl-3 methylimidazoium chloride (10), 1-octadecyl-3-methylimidazolium chloride (11), 1 octadecyl-3-methylimidazolium hexafluorophosphate (12), 1-octadecyl-3 methylimidazolium bis(triflic)imide (13) and 1-octadecyl-3-methylimidazolium tris(pentafluoroethyl)trifluorophosphate (14) were highly active against leukemia cell lines, especially compounds 13 and 14 where the cytotoxicity was also very low as given by LC(50) >100microM in all six leukemia cell lines. PMID- 20006502 TI - Ca2+ sensitizers: An emerging class of agents for counterbalancing weakness in skeletal muscle diseases? AB - Ca(2+) ions are key regulators of skeletal muscle contraction. By binding to contractile proteins, they initiate a cascade of molecular events leading to cross-bridge formation and ultimately, muscle shortening and force production. The ability of contractile proteins to respond to Ca(2+) attachment, also known as Ca(2+) sensitivity, is often compromised in acquired and congenital skeletal muscle disorders. It constitutes, undoubtedly, a major physiological cause of weakness for patients. In this review, we discuss recent studies giving strong molecular and cellular evidence that pharmacological modulators of some of the contractile proteins, also termed Ca(2+) sensitizers, are efficient agents to improve Ca(2+) sensitivity and function in diseased skeletal muscle cells. In fact, they compensate for the impaired contractile proteins response to Ca(2+) binding. Currently, such Ca(2+) sensitizing compounds are successfully used for reducing problems in cardiac disorders. Therefore, in the future, under certain conditions, these agents may represent an emerging class of agents to enhance the quality of life of patients suffering from skeletal muscle weakness. PMID- 20006503 TI - Control of mitochondrial transport and localization in neurons. AB - Mitochondria play an essential role in ATP generation, calcium buffering and apoptotic signalling. In neurons, the transport of mitochondria to specific locations where they are needed has emerged as an important process for correct nerve cell function. Recent studies have shed light on the mechanisms that control mitochondrial transport and localization in neurons. We describe the machinery that is important for constitutive transport of mitochondria throughout the cell, and highlight recent advances in our understanding of how signalling pathways can converge on this machinery and allow for rapid activity-dependent control of mitochondrial trafficking and localization. Regulation of mitochondrial trafficking might work in concert with mitochondrial tethering systems to give precise control of mitochondrial delivery and localization to regions of high energy and calcium buffering requirements within neurons. PMID- 20006504 TI - Deep tissue injury rat model for pressure ulcer research on spinal cord injury. AB - Many rat/mouse pressure ulcer (PU) models have been developed to test different hypotheses to gain deeper understanding of various causative risk factors, the progress of PUs, and assessing effectiveness of potential treatment modalities. The recently emphasized deep tissue injury (DTI) mechanism for PU formation has received increased attention and several studies reported findings on newly developed DTI animal models. However, concerns exist for the clinical relevance and validity of these models, especially when the majority of the reported rat PU/DTI models were not built upon SCI animals and many of the DTI research did not simulate well the clinical observation. In this study, we propose a rat PU and DTI model which is more clinically relevant by including chronic SCI condition into the rat PU model and to simulate the role of bony prominence in DTI formation by using an implant on the bone-tissue interface. Histological data and imaging findings confirmed that the condition of chronic SCI had significant effect on pressure induced tissue injury in a rat PU model and the including a simulated bony prominence in rat DTI model resulted in significantly greater injury in deep muscle tissue. Further integration of the SCI condition and the simulated bony prominence would result a rat PU/DTI model which can simulate even more accurately the clinical phenomenon and yield more clinically relevant findings. PMID- 20006505 TI - Differences in self-selected and fastest-comfortable walking in post-stroke hemiparetic persons. AB - Post-stroke hemiparetic walking is typically asymmetric. Assessment of symmetry is often performed at either self-selected or fastest-comfortable walking speeds to gain insight into coordination deficits and compensatory mechanisms. However, how walking speed influences the level of asymmetry is unclear. This study analyzed relative changes in paretic and non-paretic leg symmetry to assess whether one speed is more effective at highlighting asymmetries in hemiparetic walking and whether there is a systematic effect of speed on asymmetry. Forty-six subjects with chronic hemiparesis walked at their self-selected and fastest comfortable speeds on an instrumented split-belt treadmill. Relative proportions (paretic leg value/(paretic+non-paretic leg value)) were computed at each speed for step length (PSR), propulsion (PP), and joint moment impulses at the ankle and hip. Thirty-six subjects did not change their step length symmetry with speed, while three subjects changed their step length values toward increased asymmetry and seven changed toward increased symmetry. Propulsion symmetry did not change uniformly with speed for the group, with 15 subjects changing their propulsion values toward increased asymmetry while increasing speed from their self-selected to fastest-comfortable and 11 decreasing the asymmetry. Both step length and propulsion symmetry were correlated with ankle impulse proportion at self-selected and fastest-comfortable speed (cf., hip impulse proportion), but ratios (self-selected value/fastest-comfortable value) of the proportion measures (PSR and PP) showed that neither step length nor propulsion symmetry correlated with the ankle impulse proportions. Thus, the individual kinetic mechanisms used to increase speed could not be predicted from PSR or PP. PMID- 20006506 TI - The perforating branches of the P1 segment of the posterior cerebral artery. AB - The perforating branches of the P1 segment of the posterior cerebral artery are vulnerable to injury. Because of their close proximity to the basilar artery, the vulnerability occurs especially during surgical interventions for vascular pathologies such as basilar apex aneurysms. Therefore, extensive knowledge of the microsurgical anatomy of this area is mandatory to prevent poor post-operative outcomes. We microscopically examined 28 P1 segments obtained from 14 adult fresh cadaver brains (6 silicone injected, 8 freshly examined). The P1 segments ranged between 2.8mm and 12.2mm (mean 6.8mm) in length with a mean outer diameter of 1.85 mm (range 0.8-4.5mm). All 94 thalamoperforating branches identified in 27 P1 segments (mean 3.35 branches per segment) arose from the postero-superior aspect of P1 and were the most proximally originating branch in nearly all specimens (96.4%). In addition in 28 P1s, 12 short circumflex arteries (42.8%; mean 0.42 branches per segment), 16 long circumflex arteries (57.1%; mean 0.57 branches per segment) and 10 medial posterior choroidal arteries (35.7%; mean 0.35 branches per segment) were identified and all originated from the posterior or postero inferior surface of the P1 segment. When the P1 segment had more than one type of branch, it was the short circumflex arteries that were always more proximal in origin than the others. The medial posterior choroidal arteries were always more distal in origin. All three branches were not observed together in any of the P1 segments. The findings in this, and future, anatomical studies may help to reduce the post-surgical morbidity and mortality rates after surgery for posterior circulation aneurysms. PMID- 20006507 TI - Five years' experience at a single centre of craniocerebral injury from winnowing fan blades. AB - India, an agriculture-based country, relies heavily on indigenous farm machinery. In our study we present 31 patients with winnowing fan blade head injury, operated on at our centre between 2004 and 2008. The mechanism and nature of the injuries, operative methods, outcome and methods of prevention are discussed with special reference to the occurrence of this type of injury in the pediatric population. PMID- 20006508 TI - Morphometry of the thoracolumbar vertebrae in sickle cell disease. AB - Patients with sickle cell disease (SCD) who have deformities and vertebral fractures due to osteoporosis may require surgery. Spinal surgeons must become familiar with the vertebral morphometry of patients with SCD and to that aim we have examined the morphometry of the thoracolumbar spine in these patients. A cohort of 100 patients with SCD was examined using plain thoraco lumbar anteroposterior/lateral radiographs and dual energy X-ray absorptiometry (DEXA). Vertebral morphometry (vertebral body diameters, pedicle, spinal canal and deformity) was assessed for different age groups. Results were compared to published studies of healthy subjects. The vertebral dimensions for the 16-20 year and the 21+-year-old groups were significantly smaller for females than males at most spinal levels, while measurements in the 6-10 years and 11-15 years age groups were similar across both sexes at most levels. No significant statistical difference was found between the diameters of the right and left pedicles. With the exception of the sagittal diameter, most of the dimensions of the vertebral bodies measured in SCD patients were less than those of healthy individuals; multiple deformities were also observed. Low bone density was noted in 32 patients. Our data highlight the differences in vertebral bone mineral density, anatomy and deformities in patients with SCD compared to healthy individuals. When considering surgical intervention for patients with SCD, it is important that pre-operative radiography and planning is undertaken, and that the surgeon is familiar with the geometry of the pedicles of the thoracolumbar vertebrae necessary for the safe insertion of pedicle screws. Osteoporosis must be considered when planning surgical interventions in these patients. PMID- 20006509 TI - Continuous intraoperative electromyographic and transcranial motor evoked potential recordings in spinal stenosis surgery. AB - Spinal stenosis refers to narrowing of the spinal canal with encroachment of the neural structures by adjacent bone and soft tissue. Surgical treatment usually offers greater pain relief and functional recovery than non-surgical treatment. Nevertheless, neurological complications from decompressive laminectomy have been reported to range between 1% and 33%. Therefore, the purpose of this prospective study was to evaluate the efficacy of transcranial motor evoked potentials (TcMEP) and continuous electromyography (EMG) to prevent irreversible pyramidal tract damage during decompressive laminectomy. We prospectively evaluated 25 patients (11 males and 14 females) who underwent decompressive laminectomy for lumbar spinal stenosis. TcMEP and EMG were monitored intraoperatively. Postoperatively all patients had regular follow-up examinations. Electrophysiological monitoring was not performed in two patients because of the use of incompatible anaesthetic regimens. In 17/25 patients there was an increase in TcMEP amplitudes of more than 50%, whereas in six patients the amplitudes only slightly increased or remained unchanged. The 17 patients with the increased TcMEP amplitudes had the greatest improvement 3 and 12 months postoperatively, based on neurological examination and the visual analog scale pain ratings (p<0.001). Intraoperative monitoring may allow rapid identification of potential damage of the neural structures and avoidance through corrective action. TcMEP and continuous EMG monitoring is an effective method for monitoring neural function cord during surgical decompression of the lumbar spine and may additionally give prognostic information for the assessment of patient outcome. PMID- 20006510 TI - Harvey Cushing and some Australian connections: part 1 - early life and work. AB - Before World War 1 both the United States of America and Australia had rather rudimentary medical systems. Enterprising practitioners in both countries tended to look to continental Europe (especially Germany) and the United Kingdom as places of medical renown. The outbreak of World War 1 changed this but also enabled doctors from both the United States and Australia to work in military hospitals in Europe. The interactions that occurred were important for their professional development and led to some close associations between American and antipodean doctors. Examples of these were the experiences of Harvey Cushing, a burgeoning American neurosurgeon, which he recounted in his diary. His commentary merits close inspection and analysis. An account of some of Cushing's Australian connections post World War 1 is given in Part 2 (in press). PMID- 20006511 TI - Suppressed neutrophil respiratory burst in patients with haemorrhagic stroke. AB - The pathophysiology underlying the high incidence of post-stroke infectious complications has not been fully understood. We measured the respiratory burst of neutrophils as an index of their bactericidal function in patients with haemorrhagic stroke, and we also measured the plasma concentrations of noradrenalin, cortisol and neutrophil function-related amino acids. Blood samples were obtained from patients who underwent emergency craniotomies for haemorrhagic stroke (n=11) or CT-guided stereotaxic aspiration of intracerebral haematomas (n=6). Neutrophils were isolated, and their chemiluminescence response to N formyl-methionyl-leucyl-phenylalanine was measured. Healthy volunteers served as controls (n=15). In patients with stroke, the chemiluminescence response of the isolated neutrophils was significantly lower than in the controls, and it was significantly inversely correlated with the plasma noradrenalin concentration. By Day 30, this value approached control levels. Other parameters measured were not significantly correlated with the chemiluminescence response. Stroke-induced suppression of the neutrophil respiratory burst may be responsible for frequent post-stroke infectious complications. PMID- 20006512 TI - Superficial siderosis of the central nervous system manifested with seizures. AB - Superficial siderosis of the central nervous system (CNS) is caused by repeated hemorrhage into the subarachnoid space with resultant hemosiderin deposition in the subpial layers of the brain and spinal cord. Progressive ataxia and hearing impairment are the most common manifestations of the disorder. We describe a male patient admitted to our hospital for recurrent seizures over the past 3 months. He was diagnosed eventually as having superficial siderosis of the CNS through brain MRI; however, he presented with only mild hearing impairment without any cerebellar deficits. Seizures can be the major presentation of the superficial siderosis. PMID- 20006513 TI - Glycolysis: a bioenergetic or a survival pathway? AB - Following inhibition of mitochondrial respiration neurons die rapidly, whereas astrocytes utilize glycolytically-generated ATP to increase their mitochondrial membrane potential, thus becoming more resistant to pro-apoptotic stimuli. Neurons are unable to increase glycolysis due to the lack of activity of the glycolysis-promoting enzyme 6-phosphofructo-2-kinase/fructose 2,6-bisphosphatase, isoform 3 (PFKFB3). In neurons, PFKFB3 is degraded constantly via the E3 ubiquitin ligase anaphase-promoting complex/cyclosome (APC/C)- CDH1. Glucose metabolism in neurons is directed mainly to the pentose phosphate pathway, leading to regeneration of reduced glutathione. In addition to their relevance to brain physiology and pathophysiology, these observations suggest that APC/C-CDH1 might link activation of glycolysis and cell proliferation as it is also involved in the regulation of cell cycle proteins. PMID- 20006515 TI - Novel and selective DNA methyltransferase inhibitors: Docking-based virtual screening and experimental evaluation. AB - The DNA methyltransferase (DNMT) enzyme family consists of four members with diverse functions and represents one of the most promising targets for the development of novel anticancer drugs. However, the standard drugs for DNMT inhibition are non-selective cytosine analogues with considerable cytotoxic side effects that have been developed several decades ago. In this work, we conducted a virtual screening of more than 65,000 lead-like compounds selected from the National Cancer Institute collection using a multistep docking approach with a previously validated homology model of the catalytic domain of human DNMT1. Experimental evaluation of top-ranked molecules led to the discovery of novel small molecule DNMT1 inhibitors. Virtual screening hits were further evaluated for DNMT3B inhibition revealing several compounds with selectivity towards DNMT1. These are the first small molecules reported with biochemical selectivity towards an individual DNMT enzyme capable of binding in the same pocket as the native substrate cytosine, and are promising candidates for further rational optimization and development as anticancer drugs. The availability of enzyme selective inhibitors will also be of great significance for understanding the role of individual DNMT enzymes in epigenetic regulation. PMID- 20006514 TI - The on-off switches of the mitochondrial uncoupling proteins. AB - Mitochondrial uncoupling proteins disengage substrate oxidation from ADP phosphorylation by dissipating the proton electrochemical gradient that is required for ATP synthesis. In doing this, the archetypal uncoupling protein, UCP1, mediates adaptive thermogenesis. By contrast, its paralogues UCP2 and UCP3 are not thought to mediate whole body thermogenesis in mammals. Instead, they have been implicated in a variety of physiological and pathological processes, including protection from oxidative stress, negative regulation of glucose sensing systems and the adaptation of fatty acid oxidation capacity to starving. Although much work has been devoted to how these proteins are activated, little is known of the mechanisms that reverse this activation. PMID- 20006516 TI - Synthesis and anti-viral activity of azolo-adamantanes against influenza A virus. AB - Chemotherapy and chemoprophylaxis of influenza is one of the most important directions of health protection activity. Due to the high rate of drug-resistant strains of influenza virus, there is a need for the search and further development of new potent antivirals against influenza with a broad spectrum of activity. In the present study, a set of di-, tri- and tetrazole derivatives of adamantane was efficiently prepared and their anti-influenza activities evaluated against rimantadine-resistant strain A/Puerto Rico/8/34. In general, derivatives of tetrazole possessed the highest virus-inhibiting activity. We demonstrated that several compounds of this set exhibited much higher activity than the currently used antiviral rimantadine, a compound of related structure. Moreover, we showed that these azolo-adamantanes were significantly less toxic. This study demonstrates that influenza viruses can be inhibited by adamantyl-azoles and thus have potential for developing antiviral agents with an alternate mechanism of action. PMID- 20006517 TI - Synthesis of alpha-tocohexaenol (alpha-T6) a fluorescent, oxidatively sensitive polyene analogue of alpha-tocopherol. AB - A polyunsaturated analogue of alpha-tocopherol was synthesized that is both fluorescent and sensitive to peroxidative chemistry that occurs in phospholipid membranes. alpha-Tocohexaenol 1, [(S)-2,5,7,8-tetramethyl-2-((1E/Z,3E,5E,7E,9E) 4,8,12-trimethyltrideca-1,3,5,7,9,11-hexaenyl)chroman-6-ol, alpha-T6] was prepared by condensing a known triene fragment triphenyl-(2,6-dimethyl-octa-2,4,6 trienoic acid methyl ester)-phosphonium bromide with a protected chromanol aldehyde, (2S)-6-{[tert-butyl(dimethyl)silyl]oxy}-2,5,7,8-tetra-methyl-3,4 dihydro-2H-chromene-2-carbaldehyde. The full side chain was then completed with isopentyl(tri-n-butyl)phosphonium bromide to give 1. The geometry of the C1'-C2' alkene appears to be Z (cis) although the coupling constants of the olefinic protons are intermediate between values normally assigned to E and Z-isomers. In ethanol, alpha-T6 has a maximum absorption at 368nm with an absorption coefficient of 45,000M(-1) cm(-1), and displays a maximum fluorescence emission at 523nm. The susceptibility of alpha-T6 to peroxidative chemistry was dependent on the concentration of azo-initiators of lipid oxidation in acetonitrile solution as well as in phospholipid vesicles. A loss of fluorescence at 520nm was observed when alpha-T6 (vesicles or alpha-T6-lipid mixtures) was exposed to peroxidative conditions, and this loss mirrored the production of conjugated dienes and trienes during the peroxidation of bulk phospholipids. Addition of natural alpha-tocopherol during the AMVN induced oxidation of 4microM alpha-T6 and 0.5mg/ml soybean PC induced a characteristic lag phase, after which the fluorescence of alpha-T6 began to lessen. Thus, alpha-T6 may be a useful reporter not only of tocopherol location in cells, but also of the extent of peroxidative events. PMID- 20006518 TI - A diazirine-based photoaffinity etoposide probe for labeling topoisomerase II. AB - Etoposide is a widely used anticancer drug that targets topoisomerase II, an essential nuclear enzyme. However, despite the fact that it has been in use and studied for more than 30years the specific site on the enzyme to which it binds is unknown. In order to identify the etoposide binding site(s) on topoisomerase II, a diazirine-based photoaffinity etoposide analog probe has been synthesized and its photoreactivity and biological activities have been characterized. Upon UV irradiation, the diazirine probe rapidly produced a highly reactive carbene species that formed covalent adducts containing stable carbon-based bonds indicating that it should also be able to form stable covalent adducts with amino acid residues on topoisomerase II. The human leukemia K562 cell growth and topoisomerase II inhibitory properties of the diazirine probe suggest that it targets topoisomerase II in a manner similar to etoposide. The diazirine probe was also shown to act as a topoisomerase II poison through its ability to cause topoisomerase IIalpha-mediated double-strand cleavage of DNA. Additionally, the diazirine probe significantly increased protein-DNA covalent complex formation upon photoirradiation of diazirine probe-treated K562 cells, as compared to etoposide-treated cells. This result suggests that the photoactivated probe forms a covalent adduct with topoisomerase IIalpha. In conclusion, the present characterization of the chemical, biochemical, and biological properties of the newly synthesized diazirine-based photoaffinity etoposide analog indicates that use of a proteomics mass spectrometry approach will be a tractable strategy for future identification of the etoposide binding site(s) on topoisomerase II through covalent labeling of amino acid residues. PMID- 20006519 TI - A boronic acid chalcone analog of combretastatin A-4 as a potent anti proliferation agent. AB - Chalcones represent a class of natural products that inhibits tubulin assembly. In this study we designed and synthesized boronic acid analogs of chalcones in an effort to compare biological activities with combretastatin A-4, a potent inhibitor of tubulin polymerization. Systematic evaluation of the positional effects of the carbonyl moiety towards inhibition of tubulin polymerization, cancer cell proliferation and angiogenesis revealed that placement of the carbonyl adjacent to the trimethoxybenzene A-ring resulted in more active compounds than when the carbonyl group was placed adjacent to the C-ring. Our study identified a boronic acid chalcone with inhibition towards 16 human cancer cell lines in the 10-200nM range, and another three cell lines with GI(50)-values below 10nM. Furthermore, this drug has significant anti-angiogenesis effects demonstrated by HUVEC tube formation and aortic ring assay. PMID- 20006520 TI - First results in the development of an on-line digital counting platform dedicated to primary measurements. AB - This paper describes the first step in the development of a digital platform dedicated to primary radionuclide measurements. In order to validate the implementation of an on-line processing based on extendable dead-times, a FPGA based digital system has been programmed for liquid scintillation counting. After a description of the hardware, TDCR (Triple to Double Coincidence Ratio) measurements are presented. Comparisons of activity measurements with the classical system operating at LNHB are used to validate the new system; coincidence counting differences observed when increasing the resolving time in the digital platform are discussed. PMID- 20006521 TI - National comparison of 131I measurement among nuclear medicine clinics of eight countries. AB - A generally applicable protocol for organizing comparisons among nuclear medicine clinics created within the IAEA project CRP E2.10.05 was tested in Brazil, Cuba, Czech Republic, India, Iran, Republic of Korea, Romania and Turkey in 2007. Comparisons of measurement of (131)I were organized by local pilot laboratories with different backgrounds and levels of experience in this field. The results and experiences gained were compared and analyzed. A majority of results in each national comparison were within 10% of the reference value. PMID- 20006522 TI - Effects of two neuromuscular fatigue protocols on landing performance. AB - The purpose of the study was to investigate the effects of two fatigue protocols on landing performance. A repeated measures design was used to examine the effects of fatigue and fatigue protocol on neuromuscular and biomechanical performance variables. Ten volunteers performed non-fatigued and fatigued landings on two days using different fatigue protocols. Repeated maximum isometric squats were used to induce fatigue on day one. Sub-maximum cycling was used to induce fatigue on day two. Isometric squat maximum voluntary contraction (MVC) was measured before and after fatigued landings on each day. During the landings, ground reaction force (GRF), knee kinematics, and electromyographic (EMG) data were recorded. Isometric MVC, GRF peaks, loading rates, impulse, knee flexion at contact, range of motion, max angular velocity, and EMG root mean square (RMS) values were compared pre- and post-fatiguing exercise and between fatigue protocols using repeated ANOVA. Fatigue decreased MVC strength (p0.05), GRF second peak, and initial impulse (p0.01), but increased quadriceps medium latency stretch reflex EMG activity (p0.012). Knee flexion at contact was 5.2 degrees greater (p0.05) during fatigued landings following the squat exercise compared to cycling. Several variables exhibited non-significant but large effect sizes when comparing the effects of fatigue and fatigue protocol. In conclusion, fatigue alters landing performance and different fatigue protocols result in different performance changes. PMID- 20006523 TI - Unexpected air in the left ventricle after aortic cannulation in two patients with severe aortic insufficiency: possible mechanisms and clinical implications. PMID- 20006524 TI - Measuring the spatial distribution of the metaattentional spotlight. AB - Studies in cognitive psychology have shown that the deployment of visual attention operates under spatial limitations, rendering its assignment to multiple locations difficult or costly. This study explored whether this conventional understanding applies to human metaattention as well. I measured the spatial distribution of metaattention during viewing of natural scenes and found that participants believed they could attend to multiple locations simultaneously. Study 2 tested whether this tendency could be modified by information about the tendency to overestimation. After participants were informed of this tendency toward overestimation with both verbal instruction and demonstrations of attentional blindness and blindness to these phenomena, the selectivity of metaattention increased. Study 3 demonstrated that participants overestimated their attentional abilities by comparing the metaattentional drawings and the actual behavioral performances of the same participants. These results were consistent with recent findings of metaattentional overestimation in change detection and suggested human insensitivity in monitoring attentional limitations. PMID- 20006525 TI - A comparison of death anxiety and quality of life of patients with advanced cancer or AIDS and their family caregivers. AB - As an existential crisis, death anxiety may create suffering and impact quality of life. Based on a sample of 101 patients (63 patients with AIDS and 38 with advanced cancer) and 79 family caregivers (43 AIDS patients' caregivers and 36 cancer patients' caregivers), this study examined the death anxiety of patients with advanced cancer and patients with AIDS and that of their family caregivers and the relationship of death anxiety and quality of life. The results indicated that AIDS patients expressed greater death anxiety than cancer patients, but death anxiety was not different among family caregivers. Both AIDS and cancer patients experienced greater death anxiety than their caregivers. Greater death anxiety was associated with lower quality of life, particularly in the psychological domain for AIDS patients. There were significant correlations between the death anxiety subscales and the quality of life subscales for family caregivers, especially for AIDS caregivers. Interventions that lessen death anxiety may enhance quality of life as death approaches, particularly for AIDS patients and their family caregivers. PMID- 20006526 TI - Treatment of coracoid process fractures associated with acromioclavicular dislocation using clavicular hook plate and coracoid screws. PMID- 20006527 TI - Epilepsy in Ireland: towards the primary-tertiary care continuum. AB - BACKGROUND: Epilepsy is a chronic neurological disease affecting people of every age, gender, race and socio-economic background. The diagnosis and optimal management relies on contribution from a number of healthcare disciplines in a variety of healthcare settings. OBJECTIVE: To explore the interface between primary care and specialist epilepsy services in Ireland. METHODS: Using appreciative inquiry, focus groups were held with healthcare professionals (n=33) from both primary and tertiary epilepsy specialist services in Ireland. RESULTS: There are significant challenges to delivering a consistent high standard of epilepsy care in Ireland. The barriers that were identified are: the stigma of epilepsy, unequal access to care services, insufficient human resources, unclear communication between primary-tertiary services and lack of knowledge. Improving the management of people with epilepsy requires reconfiguration of the primary tertiary interface and establishing clearly defined roles and formalised clinical pathways. Such initiatives require resources in the form of further education and training and increased usage of information communication technology (ICT). CONCLUSION: Epilepsy services across the primary-tertiary interface can be significantly enhanced through the implementation of a shared model of care underpinned by an electronic patient record (EPR) system and information communication technology (ICT). Better chronic disease management has the potential to halt the progression of epilepsy with ensuing benefits for patients and the healthcare system. PMID- 20006528 TI - Randomised clinical trial of foam sclerotherapy for patients with a venous leg ulcer. AB - OBJECTIVES: To assess whether routine use of foam sclerotherapy, in addition to four-layer compression bandaging, could speed up the healing of venous ulcers. DESIGN: Randomised controlled trial involving patients recruited from a nurse-led leg ulcer clinic. A total of 315 new patients were assessed, and eleven patients were identified from follow-up clinics. METHODS: Inclusion criteria were: patients with an active venous leg ulcer, in the presence of superficial truncal venous incompetence and without total deep venous incompetence on duplex imaging. Patients were randomised to four-layer compression bandages alone (control) or with additional foam sclerotherapy to incompetent superficial truncal veins. The primary endpoint was ulcer healing 24 weeks after randomisation. RESULTS: It was only possible to recruit 40 patients who were suitable for analysis: 22 control, 18 additional foam sclerotherapy. There was no complication from the foam treatment and at six months the target vein was occluded in 9 of 11 evaluable patients that had foam. One patient died before 24 weeks from an unrelated cause. At 24 weeks, 17 of 20 (85% - 1 died) in the control group and 12 of 13 (92%) patients with additional foam sclerotherapy had ulcer healing (P=0.72, log rank testing). CONCLUSION: This trial failed to recruit sufficient patients for formal comparison, but foam sclerotherapy was feasible as an adjunct to compression therapy for venous ulceration. TRIAL REGISTRATION: Eudra CT 2005-001551-38. PMID- 20006529 TI - A paramagnetic implant containing lithium naphthalocyanine microcrystals for high resolution biological oximetry. AB - Lithium naphthalocyanine (LiNc) is a microcrystalline EPR oximetry probe with high sensitivity to oxygen [R.P. Pandian, M. Dolgos, C. Marginean, P.M. Woodward, P.C. Hammel, P.T. Manoharan, P. Kuppusamy, Molecular packing and magnetic properties of lithium naphthalocyanine crystal: hollow channels enabling permeability and paramagnetic sensitivity to molecular oxygen J. Mater. Chem. 19 (2009) 4138-4147]. However, direct implantation of the crystals in the tissue for in vivo oxygen measurements may be hindered by concerns associated with their direct contact with the tissue/cells and loss of EPR signal due to particle migration in the tissue. In order to address these concerns, we have developed encapsulations (chips) of LiNc microcrystals in polydimethyl siloxane (PDMS), an oxygen-permeable, bioinert polymer. Oximetry evaluation of the fabricated chips revealed that the oxygen sensitivity of the crystals was unaffected by encapsulation in PDMS. Chips were stable against sterilization procedures or treatment with common biological oxidoreductants. In vivo oxygen measurements established the ability of the chips to provide reliable and repeated measurements of tissue oxygenation. This study establishes PDMS-encapsulated LiNc as a potential probe for long-term and repeated measurements of tissue oxygenation. PMID- 20006530 TI - Trends from 1995 to 2006 in the prevalence of self-reported cardiovascular risk factors among elderly Spanish diabetics. AB - AIM: In this study, we examined the trends from 1995 to 2006 in cardiovascular risk factors (CVRF) in Spaniards aged 65 years or older with diabetes. METHODS: We looked at the individual data from adults aged > or =65 years from the Spanish National Health Surveys of 1995 (n=1117), 1997 (n=1111), 2001 (n=4328), 2003 (n=6134) and 2006 (n=7835). Those classified as having diabetes had answered the two following questions in the affirmative: Has your doctor told you that you currently have diabetes? Have you taken any medication to treat diabetes in the last two weeks? The CVRF of interest included high blood pressure (HBP), high cholesterol levels, obesity (BMI > or =30 kg/m(2)), smoking and sedentarity, which were estimated and compared for prevalence by survey year, age group and gender. Progression over time was analyzed using logistic-regression models. RESULTS: During the study period, the prevalences of all of the CVRF of interest were significantly higher among the elderly with diabetes compared with those without diabetes, except for current smoking, which was less frequent. The percentages of diabetic patients with HBP and obesity increased from 49.6 and 17.1%, respectively, in 1995 to 64 and 30.6%, respectively, in 2006 (adjusted ORs: 1.95 for HBP; 2.22 for obesity). CONCLUSION: Overall, the self-reported prevalence of CVRF among elderly patients with diabetes did not improve during 1995-2006 but, instead, showed significant increases in self-reported obesity and HBP. This lack of improvement calls for further investigations, and the dedicated attention of both healthcare providers and the diabetic patients themselves. PMID- 20006531 TI - [Comparison of three dosimetric techniques for lung tumor irradiation]. AB - PURPOSE: Comparison of three dosimetric techniques of lung tumor delineation to integrate tumor motion during breathing. PATIENTS AND METHODS: Nineteen patients with T1-3N0M0 malignant lung tumor were treated with definitive chemoradiotherapy (14 cases) or pre-surgery chemoradiation. Doses were, respectively, 66 and 46Gy. CT-scan for delineation was performed during three phases of breathing: free breathing and deep breath-hold inspiration and expiration. GTV (gross tumor volume) was delineated on the three sequences. The classic technique included GTV from the free-breathing sequence plus a CTV (clinical target volume) margin of 5 to 8mm plus a PTV (planning target volume) margin of 7 to 10mm (including ITV [internal target volume] margin and set-up margin). The gating-like technique included GTV from the deep breath-hold inspiration sequence plus a CTV margin of 5 to 8mm plus a PTV margin of 2mm. The three-volume technique, included GTV as a result of the fusion of GTVs from the three sequences plus a CTV margin of 5 to 8mm plus a PTV margin of 2mm. Dosimetry was calculated for the three PTVs, if possible, with the same fields number and position. Dose constraints and rules were imposed to accept dosimetries: firstly spinal cord maximal dose less than 45Gy, followed by V95 % for PTV greater than or equal to 95 %, and V20 GY(Gy) for lung less than or equal to 30 %, V30 GY(Gy) for lung less than or equal to 20 %. RESULTS: GTVs were not statistically different between the three methods of delineation. PTVs were significantly lower with the gating-like technique. V95% of the PTV were not different between the three techniques. With the classic-, the gating-like- and the 3-volume techniques, dosimetry was considered as acceptable, respectively in 15, 18 and 15 cases. Comparisons of constraint values showed that the gating-like method gave the best results. In the case of pre operative management, the gating-like method allowed the best results even for the V95% values. However, in the absence of gating device or without the possibility to use it, the 3-volume method allowed to take into account more precisely the organ motion than the classical technique. CONCLUSION: The 3-volume method can be done. It is a good method to take into account the organ motions. However, the gating-like method gives the best results leading to propose its use even for pre-operative patients with upper tumors. PMID- 20006532 TI - An efficient ultrasound-promoted method for the one-pot synthesis of 7,10,11,12 tetrahydrobenzo[c]acridin-8(9H)-one derivatives. AB - A new, efficient and general method for preparation of 7,10,11,12 tetrahydrobenzo[c]acridin-8(9H)-one derivatives using ultrasound irradiation is reported. Under ultrasound, the reaction time is short, the yields are high and the reaction conditions are mild. PMID- 20006533 TI - Matricellular proteins and inflammatory cells: a task force to promote or defeat cancer? AB - In the last years it became clear that the tumor microenvironment plays a major role in neoplastic growth. Proteins secreted either by the malignant cells or by the tumor-associated stromal cells act as extracellular signal transductors, orchestrating tumor progression. Sentinel cells of the innate immune system patrol the different organs and have proven either to promote tumor growth or induce tumor suppression. In recent years, members of the matricellular family of extracellular proteins were shown to be involved in different aspects of the inflammatory response during tumor development, although in contradictory ways. In this review we discuss the evidence available up to date that relates matricellular proteins with the regulation of the inflammatory response and tumor progression. PMID- 20006534 TI - Mild versus severe stress and BVOCs: thresholds, priming and consequences. AB - Plant-generated volatile organic compounds (BVOCs) play key roles in large-scale atmospheric processes and serve the plants as important defense and signal molecules. The main emphasis in quantitative BVOC studies has been on constitutive emissions of isoprene and specific monoterpene species that are present in only certain emitting plant species. However, environmental and biotic stresses can induce emissions of an array of organic compounds in any plant species, whereas the magnitude of emissions induced by given stress depends on stress tolerance, timing, duration and severity (mild versus strong) of the stress. The main view put forward in this review is that quantitative understanding of stress effects is the key for constructing realistic models of both constitutive and induced BVOC emissions. PMID- 20006535 TI - The 'ins' and 'outs' of flavonoid transport. AB - The sites of plant flavonoid biosynthesis, storage and final function often differ at the subcellular, cell, and even tissue and organ levels. Efficient transport systems for flavonoids across endomembranes and the plasma membrane are therefore required. However, a clear picture of the dynamic trafficking of flavonoids is only now beginning to emerge and appears to have many players. Here, we review current hypotheses for flavonoid transport, discuss whether these are mutually exclusive, highlight the importance of flavonoid efflux from vacuoles to the cytosol and consider future efforts to catch flavonoids 'in the act' of moving within and between cells. An improved understanding of transport mechanisms will facilitate the successful metabolic engineering of flavonoids for plant protection and human health. PMID- 20006536 TI - Perceptual learning, motor learning, and automaticity. PMID- 20006538 TI - Acquisition of nutrients by Chlamydiae: unique challenges of living in an intracellular compartment. AB - The Chlamydiae are obligate intracellular pathogens that replicate within a membrane-bound vacuole, termed the 'inclusion'. From this compartment, bacteria acquire essential nutrients by selectively redirecting transport vesicles and hijacking intracellular organelles. Rerouting is achieved by several mechanisms including proteolysis-mediated fragmentation of the Golgi apparatus, recruitment of Rab GTPases and SNAREs, and translocation of cytoplasmic organelles into the inclusion lumen. Given Chlamydiae's extended coevolution with eukaryotic cells, it is likely that co-option of multiple cellular pathways is a strategy to provide redundancy in the acquisition of essential nutrients from the host and has contributed to the success of these highly adapted pathogens. PMID- 20006539 TI - Merkel cell polyomavirus is not detected in mesotheliomas. AB - BACKGROUND: Merkel cell polyomavirus (MCPyV) is the first polyoma virus consistently linked to the etiology of a human cancer. Serological studies indicate that the virus is commonly acquired in childhood, with seroprevalence reaching 50% or higher among young adults. The modes of MCPyV transmission are still unclear, but it has been identified in respiratory tract samples. Given its respiratory tropism, we examined whether MCPyV could be detected in mesothelioma tissue, a malignancy induced in animal models by another polyomavirus, SV40. OBJECTIVE: To determine if MCPyV DNA can be detected in mesothelioma. STUDY DESIGN: DNA was extracted from 45 fresh-frozen mesothelioma samples. PCR was used to detect and quantify the abundance of MCPyV DNA, and a human control gene, in duplicates of the tissues. DNA from a sequence verified MCC tumor was used as a positive control. RESULTS: The human control gene was detected at high levels in all but three mesothelioma tissues. MCPyV DNA was detected in only one mesothelioma, and the level of viral DNA was very low. CONCLUSIONS: These results are inconsistent with the hypothesis that MCPyV is etiologically linked to mesothelioma. PMID- 20006540 TI - The prognosis of highly active antiretroviral therapy (HAART) treated HIV infected patients in Serbia, related to the time of treatment initiation. AB - BACKGROUND: With the introduction of highly active antiretroviral treatment (HAART) an impressive improvement in patient survival and quality of life has bee observed. However, the optimal timing of initial HAART is still under consideration. OBJECTIVE: To investigate the prognosis of HAART treated patients in Serbia, related to the timing of HAART initiation. STUDY DESIGN: A series of 563 patients on HAART was retrospectively analyzed to investigate treatment response and survival. RESULTS: After a mean of 6 years (range 1-14) of treatment with PI-based and/or NNRTI-based regimens, a favorable response was achieved in 72.4%, treatment failure occurred in 7.9%, while 19.7% had a dissociative immunological/virological response. If treatment was initiated during primary HIV infection it took a shorter time to achieve a favorable response than in patients who began HAART in chronic HIV infection (2.7+/-2.2 years vs. 6.9+/-2.7 years, P<0.01). A higher proportion of patients with primary HIV infection then those treated in the chronic phase achieved a favorable response to HAART (88.4% vs. 71.9%, P=0.045). Patients who initiated HAART when their CD4 cell counts were below 200 cells/microL needed longer treatment for favorable response (8 years vs. 6 years, log rank P<0.01). Forty-seven (8.3%) patients died. The overall estimated survival was 13 years. Patients older then 40 and IVDU were more likely to die during HAART (OR 2.6, 95% CI 1.1-5.9, P=0.016, and OR 2.0, 95% CI 1.0-3.7, P=0.02, respectively). However, reaching and maintaining undetectable viremia was an independent predictor of longer survival (OR 11.3, 95% CI 4.6-27.7, P<0.01). CONCLUSION: Reaching and maintaining undetectable viremia during HAART predicted longer survival, even if sub-clinical immunodeficiency remained. PMID- 20006541 TI - Crimean-Congo hemorrhagic fever in Iran and neighboring countries. AB - Crimean-Congo hemorrhagic fever (CCHF) is a zoonotic viral disease that is asymptomatic in infected livestock, but a serious threat to humans. Human infections begin with nonspecific febrile symptoms, but progress to a serious hemorrhagic syndrome with a case fatality rate of 2-50%. Although the causative virus is often transmitted by ticks, livestock-to-human and human-to-human transmissions also occur. The disease is one of the most widely distributed viral hemorrhagic fevers occurring in Africa, the Middle East, Asia, and some parts of Europe. In this study, we have focused on the CCHF situation in Iran and neighboring countries and provide evidence of over 5000 confirmed cases of CCHF in a single period/season. PMID- 20006542 TI - Human cytomegalovirus in utero transmission: follow-up of 524 maternal seroconversions. PMID- 20006543 TI - Differences in TMS-evoked responses between schizophrenia patients and healthy controls can be observed without a dedicated EEG system. AB - OBJECTIVE: The combination of transcranial magnetic stimulation (TMS) and electroencephalography (EEG) has been hampered by the large artifact that the TMS generates in the EEG. Using TMS with EEG necessitates a sophisticated artifact resistant EEG system that can acquire reliable signals in the crucial several tens of milliseconds immediately following the TMS pulse. Here, we demonstrate the use of a novel artifact removal algorithm together with a 24-bit EEG system to achieve similar recordings as those obtained with the dedicated TMS-compatible EEG system. METHODS: This setup was used to compare TMS-evoked responses between a group of healthy controls and a group of patients with schizophrenia, a condition in which effective neural connectivity is thought to be compromised. RESULTS: We observe differences in TMS-evoked responses between the two groups, similar to those recently reported in a study that used a dedicated TMS compatible EEG system. CONCLUSIONS: The standard 24-bit EEG system combined with an artifact removal algorithm produces results similar to the dedicated TMS compatible system. SIGNIFICANCE: This paves the way for more researchers and clinicians to use TMS-evoked responses for research and diagnosis of a wide spectrum of disorders. PMID- 20006544 TI - Changes in subthalamic activity during movement observation in Parkinson's disease: is the mirror system mirrored in the basal ganglia? AB - OBJECTIVE: The observation of a voluntary movement executed by another person is associated with an alpha and beta EEG desynchronization over the motor cortex, thought to reflect activity from the human "mirror neuron" system. The aim of our work was to study the changes in local field potentials (LFP) recorded from the subthalamic nucleus (STN) and their relationship with cortical activity, during movement observation. METHODS: Bilateral EEG and STN LFP recordings were acquired in 18 patients with Parkinson's disease, through surgically implanted electrodes for deep brain stimulation. Oscillatory changes during movement execution and movement observation were compared with two different control conditions (simple stimulus and rotating stimulus observation), in "off" and "on" motor states. Time frequency transforms and event-related coherence were used for the analysis. RESULTS: Movement observation was accompanied by bilateral beta reduction in subthalamic power and cortico-STN coherence, which was smaller than the decrease observed during movement execution, but significant when compared with the two control conditions. CONCLUSIONS: Movement observation is accompanied by changes in the beta oscillatory activity of the STN, similar to those observed in the EEG. SIGNIFICANCE: These changes suggest that the basal ganglia might be engaged by the activity of the human mirror system. PMID- 20006545 TI - Independent component approach to the analysis of EEG recordings at early stages of depressive disorders. AB - OBJECTIVE: A modern approach for blind source separation of electrical activity represented by Independent Components Analysis (ICA) was used for QEEG analysis in depression. METHODS: The spectral characteristics of the resting EEG in 111 adults in the early stages of depression and 526 non-depressed subjects were compared between groups of patients and healthy controls using a combination of ICA and sLORETA methods. RESULTS: Comparison of the power of independent components in depressed patients and healthy controls have revealed significant differences between groups for three frequency bands: theta (4-7.5Hz), alpha (7.5 14Hz), and beta (14-20Hz) both in Eyes closed and Eyes open conditions. An increase in slow (theta and alpha) activity in depressed patients at parietal and occipital sites may reflect a decreased cortical activation in these brain regions, and a diffuse enhancement of beta power may correlate with anxiety symptoms playing an important role on the onset of depressive disorder. CONCLUSIONS: ICA approach used in the present study allowed us to localize the EEG spectra differences between the two groups. SIGNIFICANCE: A relatively rare approach which uses the ICA spectra for comparison of the quantitative parameters of EEG in different groups of patients/subjects allows to improve an accuracy of measurement. PMID- 20006546 TI - Breaks during 5Hz rTMS are essential for facilitatory after effects. AB - OBJECTIVE: Stimulation frequency has been considered the most important factor in conventional repetitive transcranial magnetic stimulation (rTMS) for determining the direction of after effects on corticospinal excitability. Here, we examined the functional relevance of breaks during high-frequency subthreshold rTMS for the induction of facilitatory after effects. METHODS: The after effects on corticospinal excitability of a standard 5Hz rTMS protocol in a block design were compared to a continuous rTMS protocol using the same number of pulses. In addition the effect of current direction both for rTMS and single pulse TMS was included in the study design. RESULTS: While 5Hz rTMS in a standard block design induces facilitatory after effects on corticospinal excitability, the continuous protocol does not induce facilitation but rather inhibition. In our study only rTMS using an initially posterior-anterior current direction in the brain leads to significant neuroplastic effects at all. CONCLUSIONS: Breaks during conventional high-frequency rTMS are a crucial factor determining the direction of induced neuroplastic changes. SIGNIFICANCE: These results contribute to the understanding of rTMS-induced neuroplasticity and are important for the design of rTMS protocols both for experimental and clinical studies. PMID- 20006547 TI - Occlusion of a large pulmonary arteriovenous malformation in a paediatric patient using multiple vascular plugs. AB - Pulmonary arteriovenous malformations (PAVMs) are caused by an abnormal vascular communication between a pulmonary artery and vein. They are usually congenital in nature and are associated with considerable morbidity and mortality if left untreated. Transcatheter techniques are the favoured option for PAVM occlusion in children, and large lesions can have multiple feeder vessels that may not be appreciated until selective angiography is performed. We describe the successful occlusion of a large PAVM in a paediatric patient using multiple Amplatzer vascular plugs. PMID- 20006548 TI - Dogs, arthropod-transmitted pathogens and zoonotic diseases. PMID- 20006549 TI - From clonal to sexual: a step in T. congolense evolution? AB - Although clearly demonstrated in Trypanosoma brucei, genetic exchange remains controversial in other trypanosome species. Recently, Morrison and co-workers applied a population-genetics analysis, and established the existence of mating in Trypanosoma congolense. Starting from this original discovery, we focus here on the important question of how mating is induced during the trypanosome life cycle and discuss the use of statistics to evidence this type of non-obligatory biological process. PMID- 20006550 TI - Only the good die young: a novel paradigm for mosquito control. AB - Andrew Read and colleagues have proposed that insecticides acting late in the vector lifetime are less susceptible to evolutionary pressures, thereby avoiding insecticide resistance. Such late-life acting insecticides would kill the vector before the pathogen's extrinsic cycle is complete, but allow the vector to remain reproductively active. Some examples of late-life acting insecticides are discussed. By targeting older vectors, the dangerous cohorts--those capable of transmitting the fully developed pathogen--are removed. PMID- 20006551 TI - Untangling the role of eosinophils in trichinellosis: a never-ending story. PMID- 20006552 TI - Male-biased sex ratio: why and what consequences for the genus Schistosoma? AB - Schistosomes are the cause of the most significant helminth disease of humans. Their unusual sexual biology is intriguing. Instead of being hermaphroditic, as is the rule in other trematode species, they are gonochoric. Furthermore, their mating system is considered to be monogamous, a characteristic shared by only 1% of living species, and their sex ratio is male-biased. In this paper we propose an explanation of the origin of the male-biased sex ratio in schistosomes and highlight the ecological and evolutionary consequences of this bias. We argue that schistosome gonochorism, monogamy and the biased sex ratio can be integrated into a single evolutionary scheme. PMID- 20006553 TI - Increased Th17 cells are accompanied by FoxP3(+) Treg cell accumulation and correlated with psoriasis disease severity. AB - Both T-helper 17 cells (Th17) and CD4(+)CD25(+) regulatory T cells (Treg) play important roles in the pathogenesis of psoriasis. However, the relationship between Th17 and Treg cells and their dynamic variations in psoriasis remain unclear. In this study, we found that both Th17 and FoxP3(+) Treg cells were increased in psoriasis patients both in the peripheral circulation and skin tissue lesions and were positively correlated with disease severity. The ratio of Th17 to Treg cells in skin tissue lesions was inversely correlated with PASI scores, while it was positively correlated with PASI scores in the circulation. IL-17 secretion by CD4(+) T cells was not regulated by Treg cells, even though Treg cells exhibited significant inhibition on CD4(+) T cells proliferation and IFN-gamma production. These findings provide new information regarding the association between Th17 and Treg cells, which will further our understanding of the pathogenesis of psoriasis. PMID- 20006554 TI - Lymphocyte characteristics in children with common variable immunodeficiency. AB - The diagnosis of common variable immunodeficiency (CVID) is reserved for patients who suffer from undefined B cell dysfunction. Division of the CVID population into subgroups enables research for underlying disease causes. We studied clinical features and lymphocyte characteristics in 38 children with CVID and compared them to 30 children with less severe antibody deficiencies (e.g. specific antibody deficiency combined with IgG subclass deficiency) and with 65 pediatric controls. Most pediatric immune phenotypes were comparable to adult CVID phenotypes, including a selective increase in newly formed B cells and a decrease in memory B cells and CD4(+) T cells. Eighteen percent of pediatric patients had a mutation in the TNFRSF13B gene, which requires further investigation. Finally, pediatric patients with decreased class-switched memory B cells had significantly more complications. A pediatric classification for CVID may enable prediction and early diagnosis of disease related complications and provide a framework for further etiologic research. PMID- 20006555 TI - Obstetric anaesthesia in low-resource settings. AB - Close co-operation between obstetricians and obstetric anaesthesia providers is crucial for the safety and comfort of parturients, particularly in low-resource environments. Maternal and foetal mortality is unacceptably high, and the practice of obstetric anaesthesia has an important influence on outcome. Well conducted national audits have identified the contributing factors to anaesthesia related deaths. Spinal anaesthesia for caesarean section is the method of choice in the absence of contraindications, but is associated with significant morbidity and mortality. Minimum requirements for safe practice are adequate skills, anaesthesia monitors, disposables and drugs and relevant management protocols for each level of care. The importance of current outreach initiatives is emphasised, and educational resources and the available financial sources discussed. The difficulties of efficient procurement of equipment and drugs are outlined. Guiding principles for the practice of analgesia for labour, anaesthesia for caesarean section and the management of obstetric emergencies, where the anaesthetist also has a central role, are suggested. PMID- 20006556 TI - Diurnal rhythms of spontaneous recurrent seizures and behavioral alterations of Wistar and spontaneously hypertensive rats in the kainate model of epilepsy. AB - Attention deficit hyperactivity disorder (ADHD) can coexist with epilepsy. Spontaneously hypertensive rats (SHRs) are considered to model ADHD with overactivity, impulsiveness, deficient sustained attention, and alterations in circadian autonomic profiles. The present study explored spontaneous recurrent seizures (SRSs) and behavioral diurnal activity rhythms in normotensive Wistar rats and SHRs in the kainate model of epilepsy. Rats were video monitored (24 h/3 months) to detect SRSs. SHRs manifested a lower seizure frequency during the light phase in the 8th and 10th weeks and a lower frequency of SRSs during the night phase accompanied by attenuated responses in hyperexcitability tests. Both epileptic strains were hyperactive, with lower anxiety levels, and their diurnal rhythms were abolished. Epileptic Wistar rats and SHRs exhibited less exploration during the dark phase. This study suggests that SHRs may be useful in modeling some aspects (particularly hypertension-related diurnal rhythm disturbance) of behavior associated with epilepsy. PMID- 20006558 TI - High-dose bone morphogenetic protein-induced ectopic abdomen bone growth. AB - BACKGROUND CONTEXT: Infuse [bone morphogenetic protein (BMP)] is increasingly used in spinal fusion surgery. PURPOSE: The authors report a rare complication of BMP use. STUDY DESIGN: This is a case report. PATIENT SAMPLE: A 55-year-old male underwent a thoracic T8 to the pelvis fusion for degenerative lumbar disc disease and pseudarthrosis at another institution. The procedure involved an anterior and posterior approach with the use of multiple units of BMP. METHODS: The patient presented to our institution with complaints of weight loss, pain, tenderness, and increasing solid growth in the left lower quadrant several months after his surgery. A computed tomography revealed ectopic bone growth in the retroperitoneal area and pelvis contiguous to the anterior lumbar exposure. RESULTS: The anterior wound was re-explored, and a large sheet of ectopic bone was removed from the retroperitoneal space. CONCLUSIONS: We report a rare case of extraspinal ectopic bone growth because of the use of multiple packages of BMP. PMID- 20006557 TI - Computed tomography-evaluated features of spinal degeneration: prevalence, intercorrelation, and association with self-reported low back pain. AB - BACKGROUND CONTEXT: Although the role of radiographic abnormalities in the etiology of nonspecific low back pain (LBP) is unclear, the frequent identification of these features on radiologic studies continues to influence medical decision making. PURPOSE: The primary purposes of the study were to evaluate the prevalence of lumbar spine degeneration features, evaluated on computed tomography (CT), in a community-based sample and to evaluate the association between lumbar spine degeneration features. The secondary purpose was to evaluate the association between spinal degeneration features and LBP. STUDY DESIGN: This is a cross-sectional community-based study that was an ancillary project to the Framingham Heart Study. SAMPLE: A subset of 187 participants were chosen from the 3,529 participants enrolled in the Framingham Heart Study who underwent multidetector CT scan to assess aortic calcification. OUTCOME MEASURES: Self-report measures: LBP in the preceding 12 months was evaluated using a Nordic self-report questionnaire. Physiologic measures: Dichotomous variables indicating the presence of intervertebral disc narrowing, facet joint osteoarthritis (OA), spondylolysis, spondylolisthesis, and spinal stenosis and the density (in Hounsfield units) of multifidus and erector spinae muscles were evaluated on CT. METHODS: We calculated the prevalence of spinal degeneration features and mean density of multifidus and erector spinae muscles in groups of individuals with and without LBP. Using the chi(2) test for dichotomous and t test for continuous variables, we estimated the differences in spinal degeneration parameters between the aforementioned groups. To evaluate the association of spinal degeneration features with age, the prevalence of degeneration features was calculated in four age groups (less than 40, 40-50, 50-60, and 60+ years). We used multiple logistic regression models to examine the association between spinal degeneration features (before and after adjustment for age, sex, and body mass index [BMI]) and LBP, and between all degeneration features and LBP. RESULTS: In total, 104 men and 83 women, with a mean age (+/-standard deviation) of 52.6+/-10.8 years, participated in the study. There was a high prevalence of intervertebral disc narrowing (63.9%), facet joint OA (64.5%), and spondylolysis (11.5%) in the studied sample. When all spinal degeneration features as well as age, sex, and BMI were factored in stepwise fashion into a multiple logistic regression model, only spinal stenosis showed statistically significant association with LBP, odds ratio (OR) (95% confidence interval [CI]): 3.45 [1.12-10.68]. Significant association was found between facet joint OA and low density of multifidus (OR [95% CI]: 3.68 [1.36-9.97]) and erector spinae (OR [95% CI]: 2.80 [1.10-7.16]) muscles. CONCLUSIONS: Degenerative features of the lumbar spine were extremely prevalent in this community-based sample. The only degenerative feature associated with self-reported LBP was spinal stenosis. Other degenerative features appear to be unassociated with LBP. PMID- 20006559 TI - Effect of circumferential constraint on nucleus pulposus tissue in vitro. AB - BACKGROUND CONTEXT: Degeneration of the intervertebral disc (IVD) involves structural changes in the annulus fibrosus (AF), which could alter the mechanical forces imposed on the nucleus pulposus (NP) tissue. This could contribute to degenerative changes that occur in the NP. PURPOSE: The purpose of the study was to determine whether circumferential constraint affects anabolic and catabolic gene expression, biochemical composition, and mechanical properties of NP tissue. STUDY DESIGN: Nucleus pulposus cells were isolated from bovine caudal IVD and allowed to form tissue for a period of two weeks. The effect of no, intermediate, or high circumferential constraint on biochemical composition (cellularity and proteoglycan and collagen synthesis), gene expression, and compressive mechanical properties was evaluated. RESULTS: Increasing the rigidity of circumferential constraint surrounding in vitro formed NP tissue resulted in decreased gene expression of aggrecan and type II collagen and increased expression of MMP-1 and ADAMTS-5. This was associated with decreased accumulation of extracellular matrix and a deterioration of the compressive mechanical properties of the tissue. CONCLUSIONS: As increased circumferential constraint can have a significant negative effect on the composition and quality of NP tissue and this raises the possibility that the AF may contribute to the degenerative or age-related alterations that occur in the NP. Further study in a functional spinal unit is required to validate this. PMID- 20006560 TI - Lag screw fixation of remote bilateral pedicle fractures of the fourth and fifth lumbar vertebrae after a single gunshot wound: a case report and technical pearl. AB - BACKGROUND CONTEXT: Of the injuries involving the lumbar spine, pedicle fractures are among the least common; those involving bilateral pedicles are rare. PURPOSE: The aims of the study were to provide the first documentation of bilateral pedicle fractures at two consecutive levels after a gunshot, to review the mechanism of injury, and to evaluate a nonfusion treatment option for pedicle fractures. STUDY DESIGN: This is a technical note and case report. OUTCOME MEASURE: The outcome measures were lumbar range of motion, return of motor and sensory functions, and return to normal activities. METHODS: A 20-year-old male sustained bilateral pedicle fractures at L4 and L5 with a massive dural tear, progressive neurologic deficits, and urinary incontinence. He underwent repair of the dural tear and lag screw fixation of the pedicle fractures without fusion. RESULTS: The patient had full range of motion of his lumbar spine, full strength in his lower extremities, and bladder control. CONCLUSIONS: This is the first report of bilateral multilevel lumbar pedicle fractures after a single penetrating gunshot wound. The case documents this injury pattern after a gunshot, reviews the mechanism of injury, and presents the successful application of a nonfusion treatment option. PMID- 20006561 TI - Functional interactions between nicotinic and P2X receptors in celiac ganglia neurons. AB - Here we characterized the cross-inhibitory interactions between nicotinic and P2X receptors of celiac neurons from the guinea pig by recording whole-cell currents induced by 1mM ACh (I(ACh)), 1mM ATP (I(ATP)) and by the simultaneous application of both agonists (I(ACh)(+ATP)). I(ACh) and I(ATP) were inhibited by hexamethonium (nicotinic channel blocker) and PPADS (P2X receptor antagonist), respectively. The amplitude of I(ACh)(+ATP) was equal to the current induced by the most effective agonist, indicating a current occlusion. Various observations indicate that I(ACh)(+ATP) is carried out through both nicotinic (nACh) and P2X channels: i) I(ACh)(+ATP) desensitisation kinetics were in between that of I(ACh) and I(ATP); ii) application of ATP+ACh, decreased I(ACh) and I(ATP), whereas no cross-desensitisation was observed between nACh and P2X receptors; iii) ATP did not affect I(ACh) in the presence of PPADS or after P2X receptor desensitisation; and iv) ACh did not affect I(ATP) when nACh channels were blocked with hexamethonium or after nACh receptor desensitisation. Current occlusion is not mediated by activation of metabotropic receptors as it is: i) voltage dependent (was not observed at + 5 mV); ii) present at low temperature (10 degrees C) and after inhibition of protein kinase activity (with staurosporine); and iii) absent at 30 microM ATP and 30 microM ACh (concentrations that should activate metabotropic receptors). In conclusion, current occlusion described here is similar to the previously reported myenteric neurons. This occlusion is likely the result of allosteric interactions between these receptors. PMID- 20006563 TI - Osteoclast function, bone turnover and inflammatory cytokines during infective exacerbations of cystic fibrosis. AB - BACKGROUND: Raised levels of pro-inflammatory, pro-resorptive cytokines during pulmonary infection may contribute to osteoporosis in cystic fibrosis (CF). We assessed osteoclast number and activity during infective exacerbations and examined their relationship to serum inflammatory cytokines and bone turnover markers. METHODS: Serum samples from 24 adults with CF were obtained before, during and after treatment of infection. Osteoclastic cells were generated from peripheral blood mononuclear cells and their number and activity assessed. Serum osteocalcin, type 1 collagen cross-linked N-telopeptide (NTx), interleukin-6 (IL 6), tumour necrosis factor alpha (TNFalpha), receptor activator of NFkB ligand (RANKL) and osteoprotegerin (OPG) were measured. RESULTS: Osteoclast number and activity were increased at the start of exacerbation and decreased with antibiotic therapy. Significant correlations were demonstrated between osteoclast formation and serum TNFalpha, OPG, osteocalcin and NTx and between osteoclast activity and serum IL-6 and NTx. CONCLUSIONS: The systemic response to infection is associated with increased bone resorptive activity in patients with CF. PMID- 20006562 TI - DNA polymerase beta-dependent long patch base excision repair in living cells. AB - We examined a role for DNA polymerase beta (Pol beta) in mammalian long patch base excision repair (LP BER). Although a role for Pol beta is well known in single-nucleotide BER, information on this enzyme in the context of LP BER has been limited. To examine the question of Pol beta involvement in LP BER, we made use of nucleotide excision repair-deficient human XPA cells expressing UVDE (XPA UVDE), which introduces a nick directly 5' to the cyclobutane pyrimidine dimer or 6-4 photoproduct, leaving ends with 3'-OH and 5'-phosphorylated UV lesion. We observed recruitment of GFP-fused Pol beta to focal sites of nuclear UV irradiation, consistent with a role of Pol beta in repair of UV-induced photoproducts adjacent to a strand break. This was the first evidence of Pol beta recruitment in LP BER in vivo. In cell extract, a 5'-blocked oligodeoxynucleotide substrate containing a nicked 5'-cyclobutane pyrimidine dimer was repaired by Pol beta-dependent LP BER. We also demonstrated Pol beta involvement in LP BER by making use of mouse cells that are double null for XPA and Pol beta. These results were extended by experiments with oligodeoxynucleotide substrates and purified human Pol beta. PMID- 20006564 TI - Metabolites of puerarin identified by liquid chromatography tandem mass spectrometry: similar metabolic profiles in liver and intestine of rats. AB - Puerarin is a major active ingredient of Pueraria radix. Puerarin may exert its medicinal functions in part via its metabolites. In this study, we identified these metabolites to better understand and elucidate puerarin's metabolic pathway. Puerarin was intravenously administered to rats and then metabolites in plasma samples were identified by rapid resolution liquid chromatography electrospray ionization-collision induced dissociation tandem mass spectrometry (RRLC-ESI-CID-MS/MS). Chromatography was conducted on a Zorbax SB C18 column (2.1x100 mm, 1.8 microm) at 30 degrees C, with a gradient mobile phase consisting of 0.05% formic acid and acetonitrile, a flow rate of 0.2 mL min(-1), and a total run time of 14 min. MS/MS acquisition parameters were as follows: positive ionization mode, dry gas: nitrogen, 10 L min(-1), dry temperature: 350 degrees C, nebulizer: 40 psi, capillary: -3500 V, scan range: 250-800. The autoMS, manual, or multiple reaction monitoring mode was selected as required. Two glucuronidated metabolites of puerarin (M1 and M2) were detected. M1 and M2 are presumed to be puerarin-7-O-glucuronide and puerarin-4'-O-glucuronide, respectively, and M2 likely is suspected to be the major metabolite because it represented the predominate peak. Kinetic studies of metabolites demonstrated that M1 and M2 were detected in rat plasma at 5 min after intravenous administration of puerarin, the levels of M1 and M2 then reached their peaks at 10-15 and 15-30 min, respectively. The metabolic profiles were similar in rat liver and intestine investigated by in situ liver and intestine perfusion, indicating that no metabolic regioselectivity of puerarin occurs in the two organs. PMID- 20006565 TI - HPLC determination of novel dithiolethione containing drugs and its application for in vivo studies in rats. AB - A panel of new drugs obtained by grafting a sulfurated moiety, i.e. 5-(4 hydroxyphenyl)-3H-1,2-dithiole-3-thione (ADTOH) onto existing drugs have been synthesized and their in vivo action is under preclinical evaluation. In the present paper we describe rapid HPLC methods to detect ADTOH derivatives of valproic acid (ACS2), sildenafil (ACS6), aspirin (ACS14) and diclofenac (ACS15) in plasma. These methods allow the simultaneous detection of the potential drugs and of ADTOH moiety. In the case of ACS14 the de-acetylated metabolite (ACS21) can also be concomitantly measured. The chromatographic separation was performed on a C18 column, applying a mobile phase consisting of a mixture of trifluoroacetic acid and acetonitrile. ADTOH, ACS6, ACS14, ACS21 were separated isocratically whereas ACS2 and ACS15 were separated applying gradient elution. The methods are precise and accurate, with a low quantification limit of 200 nM for ACS2, ACS15 and ACS21 or 100 nM for ADTOH, ACS6 and ACS14. The mean absolute recovery for all tested molecules was always found to be close to 100%. The methods are shown to be selective and linear in the range 0.2-50 microM and thus appear suitable for pharmacokinetic studies with ADTOH containing compounds, as indicated by exemplificative experiments performed with intravenous administration of the drugs to rats. PMID- 20006566 TI - [Isolated bacteraemia caused by Kingella kingae]. PMID- 20006567 TI - Metalloporphyrins in the management of neonatal hyperbilirubinemia. AB - Neonatal jaundice in the first week of life is a common problem in newborns. It is due to an imbalance of bilirubin production and its elimination, which can lead to significantly elevated levels of circulating bilirubin or hyperbilirubinemia. Use of phototherapy and/or exchange transfusion are the current modes for treating neonatal hyperbilirubinemia and preventing any neurologic damage. These strategies, however, only remove bilirubin that has already been formed. Preventing the production of excess bilirubin may be a more logical approach. Synthetic heme analogs, metalloporphyrins, are competitive inhibitors of heme oxygenase, the rate-limiting enzyme in bilirubin production, and their use has been proposed as an attractive alternative strategy for preventing or treating severe hyperbilirubinemia. PMID- 20006568 TI - Comparative study of the effects of aluminum adjuvants and Freund's incomplete adjuvant on the immune response to an Edwardsiella tarda major antigen. AB - Edwardsiella tarda is a severe aquaculture pathogen that can infect many different fish species cultured worldwide. Et49 is a major E. tarda antigen with weak immunoprotective potential. In this study, using Et49 as an example vaccine, the adjuvanticity of Freund's incomplete adjuvant (FIA), aluminum hydroxide, and aluminum phosphate adjuvant were evaluated in a Japanese flounder model. The results showed that the presence of FIA, aluminum hydroxide, and aluminum phosphate adjuvant increased the relative percent of survival of Et49-vaccinated fish by 47%, 19%, and 35%, respectively. Fish vaccinated with FIA-adjuvanted Et49 exhibited longer persistence of vaccine at the injection site and more severe intra-abdominal lesions than fish vaccinated with aluminum-adjuvanted Et49. Both aluminum adjuvants and, to a lesser degree, FIA augmented the production of specific serum antibodies, which reached the highest levels at 6 and 7 weeks post vaccination. Passive immunization of Japanese flounder with sera from fish vaccinated with aluminum- and FIA-adjuvanted Et49 induced no protection against lethal E. tarda challenge. Examination of the transcription profile of immune related genes showed that vaccination with aluminum-adjuvanted Et49 significantly enhanced the expression of the genes that are associated mainly with humoral immunity, whereas vaccination with FIA-adjuvanted Et49 induced the expression of a much broader spectrum of genes that are likely to be involved in humoral and innate cellular immunity. These results provide new insights to the action mechanisms of FIA and aluminum adjuvants in Japanese flounder and may be useful for the selection of adjuvant for vaccine formulations intended for Japanese flounder. PMID- 20006569 TI - Polio eradication in India: have we reached the dead end? PMID- 20006570 TI - Socioeconomic factors play a more important role in childhood vaccination coverage than parental perceptions: a cross-sectional study in Greece. AB - To identify predictive factors of complete and age-appropriate vaccination status in Greece, we conducted a cross-sectional study, using stratified cluster sampling, among children attending the first year of the Greek Grammar school (about 6 years of age) and their parents/guardians. Almost 88% (N=3878) of pupils in the selected clusters (school classrooms) provided their vaccination booklet and their parents/guardians completed a questionnaire regarding beliefs and attitudes towards immunization. Belonging to a minority group, having other siblings and perceiving long distance to immunization site as a barrier were independent predictors of both incomplete and delayed vaccination status in the final logistic regression model. Maternal age >or=30 years and the perception that natural disease is preferable to vaccination were associated with complete vaccination, whereas paternal education of high school or higher was the other independent determinant of age-appropriate immunization. Socioeconomic factors rather than parental beliefs and attitudes towards immunization explained underimmunization. Further interventions are warranted to enhance vaccine coverage in high-risk groups identified in this study. PMID- 20006572 TI - Inactivation of nitric oxide by cytochrome c oxidase under steady-state oxygen conditions. AB - We have developed a respiration chamber that allows intact cells to be studied under controlled oxygen (O(2)) conditions. The system measures the concentrations of O(2) and nitric oxide (NO) in the cell suspension, while the redox state of cytochrome c oxidase is continuously monitored optically. Using human embryonic kidney cells transfected with a tetracycline-inducible NO synthase we show that the inactivation of NO by cytochrome c oxidase is dependent on both O(2) concentration and electron turnover of the enzyme. At a high O(2) concentration (70 microM), and while the enzyme is in turnover, NO generated by the NO synthase upon addition of a given concentration of l-arginine is partially inactivated by cytochrome c oxidase and does not affect the redox state of the enzyme or consumption of O(2). At low O(2) (15 microM), when the cytochrome c oxidase is more reduced, inactivation of NO is decreased. In addition, the NO that is not inactivated inhibits the cytochrome c oxidase, further reducing the enzyme and lowering O(2) consumption. At both high and low O(2) concentrations the inactivation of NO is decreased when sodium azide is used to inhibit cytochrome c oxidase and decrease electron turnover. PMID- 20006571 TI - Human embryonic kidney (HEK293) cells express endogenous voltage-gated sodium currents and Na v 1.7 sodium channels. AB - Human embryonic kidney (HEK293) cells are widely used for the heterologous expression of voltage- and ligand-gated ion channels. Patch clamp analysis of HEK293 cells in the whole-cell configuration identified voltage-gated, rapidly inactivating inward currents. Peak current amplitudes ranged from less than 100 pA to more than 800 pA, with the majority (84 of 130 cells) in the 100-400 pA range. Transient inward currents were separated into three components on the basis of sensitivity to cadmium and tetrodotoxin (TTX). Application of cadmium (300 microM) reduced current amplitude to 65% of control, consistent with the existence of current carried by a cadmium-sensitive nonspecific cation channel previously identified in HEK293 cells. Application of TTX (500 nM) reduced current amplitude by 47%, consistent with the existence of current carried by a TTX-sensitive voltage-gated sodium channel. Joint application of cadmium and TTX was additive, reducing current amplitude to 28% of control. The residual cadmium- and TTX-resistant currents represent a third pharmacologically distinct component of the rapidly inactivating inward current that was not characterized further. The pyrethroid insecticide tefluthrin (10 microM) prolonged the inactivation of transient currents and induced slowly decaying tail currents, effects that are characteristic of sodium channel modification by pyrethroids. The use of sodium channel isoform-specific primers in polymerase chain reaction amplifications on HEK293 cell first-strand cDNA detected the consistent expression of the human Na(v)1.7 sodium channel isoform in cells that expressed the TTX-sensitive component of current. These results provide evidence for an endogenous TTX sensitive sodium current in HEK293 cells that is associated primarily with the expression of the Na(v)1.7 sodium channel isoform. PMID- 20006573 TI - Tumor Necrosis Factor alpha (TNFalpha) regulates CD40 expression through SMAR1 phosphorylation. AB - CD40 plays an important role in mediating inflammatory response and is mainly induced by JAK/STAT phosphorylation cascade. TNFalpha is the key cytokine that activates CD40 during inflammation and tumorigenesis. We have earlier shown that SMAR1 can repress the transcription of Cyclin D1 promoter by forming a HDAC1 dependent repressor complex. In this study, we show that SMAR1 regulates the transcription of NF-kappaB target gene CD40. SMAR1 recruits HDAC1 and forms a repressor complex on CD40 promoter and keeps its basal transcription in check. Further, we show that TNFalpha stimulation induces SMAR1 phosphorylation at Ser 347 and promotes its cytoplasmic translocation, thus releasing its negative effect. Concomitantly, TNFalpha induced phosphorylation of STAT1 at Tyr-701 by JAK1 facilitates its nuclear translocation and activation of CD40 through p300 recruitment and core Histone-3 acetylation. Thus, TNFalpha mediated regulation of CD40 expression occurs by dual phosphorylation of SMAR1 and STAT1. PMID- 20006574 TI - The liver-enriched transcription factor CREBH is nutritionally regulated and activated by fatty acids and PPARalpha. AB - To elucidate the physiological role of CREBH, the hepatic mRNA and protein levels of CREBH were estimated in various feeding states of wild and obesity mice. In the fast state, the expression of CREBH mRNA and nuclear protein were high and profoundly suppressed by refeeding in the wild-type mice. In ob/ob mice, the refeeding suppression was impaired. The diet studies suggested that CREBH expression was activated by fatty acids. CREBH mRNA levels in the mouse primary hepatocytes were elevated by addition of the palmitate, oleate and eicosapenonate. It was also induced by PPARalpha agonist and repressed by PPARalpha antagonist. Luciferase reporter gene assays indicated that the CREBH promoter activity was induced by fatty acids and co-expression of PPARalpha. Deletion studies identified the PPRE for PPARalpha activation. Electrophoretic mobility shift assay and chromatin immunoprecipitation (ChIP) assay confirmed that PPARalpha directly binds to the PPRE. Activation of CREBH at fasting through fatty acids and PPARalpha suggest that CREBH is involved in nutritional regulation. PMID- 20006575 TI - Bone marrow-derived cells are differentially involved in pathological and physiological retinal angiogenesis in mice. AB - PURPOSE: Bone marrow-derived cells have been shown to play roles in angiogenesis. Although these cells have been shown to promote angiogenesis, it is not yet clear whether these cells affect all types of angiogenesis. This study investigated the involvement of bone marrow-derived cells in pathological and physiological angiogenesis in the murine retina. MATERIALS AND METHODS: The oxygen-induced retinopathy (OIR) model was used as a retinal angiogenesis model in newborn mice. To block the influence of bone marrow-derived cells, the mice were irradiated with a 4-Gy dose of radiation from a (137)Cs source. Irradiation was performed in four different conditions with radio dense 2-cm thick lead disks; (1) H group, the head were covered with these discs to protect the eyes from radiation; (2) A group, all of the body was covered with these discs; (3) N group, mice were completely unshielded; (4) C group, mice were put in the irradiator but were not irradiated. On P17, the retinal areas showing pathological and physiological retinal angiogenesis were measured and compared to the retinas of nonirradiated mice. RESULTS: Although irradiation induced leukocyte depletion, it did not affect the number of other cell types or body weight. Retinal nonperfusion areas were significantly larger in irradiated mice than in control mice (P<0.05), indicating that physiological angiogenesis was impaired. However, the formation of tuft-like angiogenesis processes was more prominent in the irradiated mice (P<0.05), indicating that pathological angiogenesis was intact. CONCLUSIONS: Bone marrow-derived cells seem to be differentially involved in the formation of physiological and pathological retinal vessels. Pathological angiogenesis in the murine retina does not require functional bone marrow-derived cells, but these cells are important for the formation of physiological vessels. Our results add a new insight into the pathology of retinal angiogenesis and bolster the hypothesis that bone marrow cells are involved in the pathology or severity of retinal angiogenic diseases. PMID- 20006576 TI - Anterior cruciate ligament-derived cells have high chondrogenic potential. AB - Anterior cruciate ligament (ACL)-derived cells have a character different from medial collateral ligament (MCL)-derived cells. However, the critical difference between ACL and MCL is still unclear in their healing potential and cellular response. The objective of this study was to investigate the mesenchymal differentiation property of each ligament-derived cell. Both ligament-derived cells differentiated into adipogenic, osteogenic, and chondrogenic lineages. In chondrogenesis, ACL-derived cells had the higher chondrogenic property than MCL derived cells. The chondrogenic marker genes, Sox9 and alpha1(II) collagen (Col2a1), were induced faster in ACL-derived pellets than in MCL-derived pellets. Sox9 expression preceded the increase of Col2a1 in both pellet-cultured cells. However, the expression level of Sox9 and a ligament/tendon transcription factor Scleraxis did not parallel the increase of Col2a1 expression along with chondrogenic induction. The present study demonstrates that the balance between Sox9 and Scleraxis have an important role in the chondrogenic differentiation of ligament-derived cells. PMID- 20006577 TI - The t-SNAREs syntaxin4 and SNAP23 but not v-SNARE VAMP2 are indispensable to tether GLUT4 vesicles at the plasma membrane in adipocyte. AB - SNARE proteins (VAMP2, syntaxin4, and SNAP23) have been thought to play a key role in GLUT4 trafficking by mediating the tethering, docking and subsequent fusion of GLUT4-containing vesicles with the plasma membrane. The precise functions of these proteins have remained elusive, however. We have now shown that depletion of the vesicle SNARE (v-SNARE) VAMP2 by RNA interference in 3T3-L1 adipocytes inhibited the fusion of GLUT4 vesicles with the plasma membrane but did not affect tethering of the vesicles to the membrane. In contrast, depletion of the target SNAREs (t-SNAREs) syntaxin4 or SNAP23 resulted in impairment of GLUT4 vesicle tethering to the plasma membrane. Our results indicate that the t SNAREs syntaxin4 and SNAP23 are indispensable for the tethering of GLUT4 vesicles to the plasma membrane, whereas the v-SNARE VAMP2 is not required for this step but is essential for the subsequent fusion event. PMID- 20006578 TI - Both N- and C-lobes of calmodulin are required for Ca2+-dependent regulations of CaV1.2 Ca2+ channels. AB - We investigated the concentration- and Ca(2+)-dependent effects of CaM mutants, CaM(12) and CaM(34), in which Ca(2+)-binding to its N- and C-lobes was eliminated, respectively, on the Ca(V)1.2 Ca(2+) channel by inside-out patch clamp in guinea-pig cardiomyocytes. Both CaM(12) and CaM(34) (0.7-10muM) applied with 3mM ATP produced channel activity after "rundown". Concentration-response curves were bell-shaped, similar to that for wild-type CaM. However, there was no obvious leftward shift of the curves by increasing [Ca(2+)], suggesting that both functional lobes of CaM were necessary for the Ca(2+)-dependent shift. However, channel activity induced by the CaM mutants showed Ca(2+)-dependent decrease, implying a Ca(2+) sensor existing besides CaM. These results suggest that both N- and C-lobes of CaM are required for the Ca(2+)-dependent regulations of Ca(V)1.2 Ca(2+) channels. PMID- 20006579 TI - Significance of HLA class I antibody-induced antioxidant gene expression for endothelial cell protection against complement attack. AB - It has been observed that a graft organ continues to survive and function normally even in the presence of anti-graft antibodies. However, the mechanisms behind acquirement of this condition remain unknown. Here we report that the anti HLA ligation on endothelial cells induces PI3K/AKT activation followed by antioxidant gene induction through Nrf2-mediated antioxidant-responsive element (ARE) activation. Activation of PI3K/AKT in endothelial cells by a low concentration of anti-HLA ligation enhances protection from complement attack. A real-time quantitative PCR and flow-cytometry experiment showed that ferritin H and HO-1 mRNAs were induced in a PI3K/AKT-dependent manner, while CD55 and CD59 expression were not enhanced by anti-HLA ligation. Anti-HLA ligation on endothelial cells activates ferritin H ARE and induces Nrf2 binding on its enhancer element. Finally, overexpression of Nrf2 in endothelial cells attenuates complement-mediated cytotoxicity. These experiments suggest that induction of PI3K/AKT-dependent cytoprotective genes by Nrf2 is an important mechanism to prevent complement attack. Thus, a protocol to activate this pathway would be a potential strategy for avoidance of graft rejection in transplantation. PMID- 20006580 TI - N-linked glycosylation determines cell surface expression of two-pore-domain K+ channel TRESK. AB - Within the first external loop of mouse and human TRESK subunits one or two N glycosylation consensus sites were identified, respectively. Using site directed mutagenesis and Western immunoblotting a single residue of both orthologues was found to be glycosylated upon heterologous expression. Two-electrode voltage clamp recordings from Xenopus oocytes revealed that current amplitudes of N glycosylation mutants were reduced by 80% as compared to wildtype TRESK. To investigate membrane targeting, GFP-tagged TRESK subunits were expressed in Xenopus oocytes and fluorescence intensity at the cell surface was measured by confocal microscopy. Signals of the N-glycosylation mutants were reduced by >50%, indicating that their lower current amplitudes substantially result from inadequate surface expression of the channel. PMID- 20006581 TI - Fatty acids of membrane phospholipids in Drosophila melanogaster lines showing rapid and slow recovery from chill coma. AB - We investigated the fatty acid compositions of phospholipids in Drosophila melanogaster lines showing rapid (CR), intermediate (CTL), or slow (CS) recovery from chill coma, which were established by artificial selection or by free recombination without selection. Compared to CTL, CS showed a low composition of dienoic acids and a small number of double bonds in the fatty acids. The ratio of unsaturated fatty acids and saturated fatty acids (UFAs/SFAs) was significantly lower in CS than in CTL. CR had higher monoenoic acid composition and lower dienoic acid composition than CTL. In addition, the amount of SFAs was lower and therefore the UFAs/SFAs ratio considerably higher in CR than in CTL. These changes in phospholipid fatty acids probably contributed to losing and maintaining the homeoviscosity of the cellular membranes in CS and CR, respectively, at low temperature and therefore produced their distinct phenotypes in recovery from chill coma. PMID- 20006582 TI - Phosphorylated hamartin-Hsp70 complex regulates apoptosis via mitochondrial localization. AB - The products of the tuberous sclerosis complex (TSC) genes, hamartin and tuberin, form a heterodimer. Recently we reported that hamartin directly interacted with Hsp70. However, the physiological implications of this interaction have not yet been clearly defined. Here we show that hamartin localized to the outer membrane of the mitochondria in an Hsp70-dependent manner. Moreover, phosphorylation of the T417 residue of hamartin was required for its localization to the mitochondria as well as its interaction with Hsp70. A non-phosphorylatable hamartin mutant at residue T417 was unable to localize to the mitochondria and suppress apoptosis, whereas non-phosphorylatable hamartin mutants T357A and T390A localized to the mitochondria and suppressed apoptosis. Importantly, non phosphorylatable mutants (T357A, T390A and T417A) promoted apoptosis after treatment with Hsp 70-inhibitor KNK437. We conclude that hamartin inhibited apoptosis by localizing to the mitochondria and that its phosphorylation and binding to Hsp70 was required for facilitation of this process. PMID- 20006583 TI - PI3K inhibition causes the accumulation of ubiquitinated presenilin 1 without affecting the proteasome activity. AB - gamma-Secretase is an enzymatic complex, composed of presenilin 1 (PS1), nicastrin, pen-2, and aph-1, and is responsible for the intramembranous cleavage of various type-I membrane proteins. The level of each component is tightly regulated in a cell via proteasomal degradation. On the other hand, it has previously been reported that PS1/gamma-secretase is involved in the activation of phosphatidylinositol-3 kinase/Akt (PI3K/Akt) pathway. PI3K is inhibited in Alzheimer's disease (AD) brain, whereas the effects of PI3K inhibition on the metabolism of PS1/gamma-secretase have not been elucidated. Here, we demonstrate that the treatment of neurons with PI3K inhibitors leads to increased levels of PS1/gamma-secretase components through an inhibitory effect on their degradation. Moreover, PI3K inhibition accelerated ubiquitination of PS1. We further show the evidence that the PS1 ubiquitination after PI3K inhibition is represented by the multiple mono-ubiquitination, instead of poly-ubiquitination. Accordingly, treatment of cells with PI3K inhibitor led to a differential intracellular redistribution of PS1 from the one observed after the proteasomal inhibition. These results suggest that PI3K inhibition may trigger the multiple mono ubiquitination of PS1, which precludes the degradation of PS1/gamma-secretase through the proteasomal pathway. Since PS1/gamma-secretase is deeply involved in the production of Abeta protein, a deeper knowledge into its metabolism could contribute to a better elucidation of AD pathogenesis. PMID- 20006584 TI - Succinimidyl oleate, established inhibitor of CD36/FAT translocase inhibits complex III of mitochondrial respiratory chain. AB - The functional role of CD36 protein detected in mitochondrial fractions in long chain fatty acid (LCFA) oxidation is unclear due to conflicting results obtained in Cd36 knockout mice and experiments using sulfo-N-succinimidyl oleate (SSO) for inhibition of CD36 mediated LCFA transport. We investigated effect of SSO on mitochondrial respiration and found that SSO substantially inhibits not only LCFA oxidation, but also oxidation of flavoprotein- and NADH-dependent substrates and generation of mitochondrial membrane potential. Experiments in rat liver, heart and kidney mitochondria demonstrated a direct effect on mitochondrial respiratory chain with the most pronounced inhibition of the complex III (IC(50) 4microM SSO). The results presented here show that SSO is a potent and irreversible inhibitor of mitochondrial respiratory chain. PMID- 20006585 TI - Identification of acrolein-conjugated protein in plasma of patients with brain infarction. AB - It is known that the level of protein-conjugated acrolein in plasma is a good marker of chronic renal failure and brain infarction. Thus, studies were carried out to determine which kinds of plasma proteins are conjugated with acrolein. It was found that acrolein was mainly conjugated with albumin. Tandem mass spectrometry analysis demonstrated that Lys-557 and Lys-560, located at the surface of domain III of albumin, were the major sites conjugated with acrolein. This is the first report to identify the amino acid residues in a protein modified by acrolein in vivo. It was found that conjugation of acrolein with albumin contributed to a decrease in the toxicity of acrolein. PMID- 20006586 TI - NEMO-binding domain peptide promotes osteoblast differentiation impaired by tumor necrosis factor alpha. AB - Osteogenesis associated with persistent inflammation or infection exists in a broad range of conditions including rheumatoid arthritis and traumatic bone fracture. The poor outcomes of these conditions will benefit from more effective treatments. Here we investigated the molecular mechanisms and tested NEMO-binding domain peptide as a new approach of circumventing TNF-alpha inhibition of osteoblast differentiation. Our results showed: TNF-alpha markedly decreased BMP 2-induced alkaline phosphatase activity in the multipotent myoblast C2C12 cells in a dose dependent manner; stepwise experiments demonstrated that BMP-2-induced Smad1 activity was abrogated by addition of exogenous TNF-alpha or overexpression of NF-kappaB, and it was significantly elevated by overexpression of IkappaBalpha, an inhibitor of NF-kappaB; Western blotting showed that TNF-alpha markedly decreased the amount of phospho-Smad1 in BMP-2-activated C2C12 cells, but it did not alter Smad1 mRNA abundance as measured by real-time PCR; addition of a functional cell-permeable NEMO-binding domain (NBD) peptide antagonized NF kappaB activity and ameliorated TNF-alpha inhibition of osteoblast differentiation. Taken together, our study reveals for the first time that NF kappaB activation inhibits osteoblast differentiation by attenuating Smad1 activity and application of NBD peptide ameliorates this inhibitory effect. This could lead to new therapeutic drugs that circumvent the inflammatory inhibition of osteogenesis for treatment of traumatic open fractures with infection, rheumatoid arthritis and other bone loss disorders. PMID- 20006587 TI - SUMOylation regulates the nuclear mobility of CREB binding protein and its association with nuclear bodies in live cells. AB - The lysine acetyltransferase CREB binding protein (CBP) is required for chromatin modification and transcription at many gene promoters. In fixed cells, a large proportion of CBP colocalises to PML or nuclear bodies. Using live cell imaging, we show here that YFP-tagged CBP expressed in HEK293 cells undergoes gradual accumulation in nuclear bodies, some of which are mobile and migrate towards the nuclear envelope. Deletion of a short lysine-rich domain that contains the major SUMO acceptor sites of CBP abrogated its ability to be SUMO modified, and prevented its association with endogenous SUMO-1/PML speckles in vivo. This SUMO defective CBP showed enhanced ability to co-activate AML1-mediated transcription. Deletion mapping revealed that the SUMO-modified region was not sufficient for targeting CBP to PML bodies, as C-terminally truncated mutants containing this domain showed a strong reduction in accumulation at PML bodies. Fluorescence recovery after photo-bleaching (FRAP) experiments revealed that YFP-CBPDelta998 1087 had a retarded recovery time in the nucleus, as compared to YFP-CBP. These results indicate that SUMOylation regulates CBP function by influencing its shuttling between nuclear bodies and chromatin microenvironments. PMID- 20006588 TI - Characterisation of the interaction between syndecan-2, neurofibromin and CASK: dependence of interaction on syndecan dimerization. AB - Neurofibromin and calcium/calmodulin-dependent serine protein kinase (CASK) are membrane-associated signalling and scaffolding proteins which are mutated in human genetic neurological disorders. Syndecan-2 is a highly glycosylated transmembrane protein whose intracellular C-terminus has previously been shown to interact with the post-synaptic density 95/discs large/zonula occludens-1 (PDZ) domain of CASK and with two separate regions of neurofibromin. These three proteins collaborate to orchestrate the induction of filopodia and dendritic spines. We have used systematic mutagenesis of the intracellular region of syndecan-2 and a quantitative yeast two-hybrid (Y2H) assay to study the determinants of their interactions. We show that syndecan's interactions with both CASK and neurofibromin are dependent on syndecan homodimerization and that neurofibromin largely interacts with the membrane-proximal part of the dimeric syndecan intracellular domain, leaving the membrane-distal C-terminus free to interact with CASK. We conducted a phylogenetic study of syndecan sequences, finding correspondence between conserved residues and mutations affecting both dimerization and interactions; we also find that fish have a very different syndecan repertoire from tetrapods. Further Y2H screens reveal that syndecan-2 interacts with a third distinct region of neurofibromin, and that the multiple neurofibromin regions bind competitively, rather than co-operatively, to syndecan. We combine these results to propose a model for the ternary syndecan neurofibromin-CASK complex. PMID- 20006589 TI - Endoplasmic reticulum calcium release potentiates the ER stress and cell death caused by an oxidative stress in MCF-7 cells. AB - Increase in cytosolic calcium concentration ([Ca2+](c)), release of endoplasmic reticulum (ER) calcium ([Ca2+](er)) and ER stress have been proposed to be involved in oxidative toxicity. Nevertheless, their relative involvements in the processes leading to cell death are not well defined. In this study, we investigated whether oxidative stress generated during ascorbate-driven menadione redox cycling (Asc/Men) could trigger these three events, and, if so, whether they contributed to Asc/Men cytoxicity in MCF-7 cells. Using microspectrofluorimetry, we demonstrated that Asc/Men-generated oxidative stress was associated with a slow and moderate increase in [Ca2+](c), largely preceding permeation of propidium iodide, and thus cell death. Asc/Men treatment was shown to partially deplete ER calcium stores after 90 min (decrease by 45% compared to control). This event was associated with ER stress activation, as shown by analysis of eIF2 phosphorylation and expression of the molecular chaperone GRP94. Thapsigargin (TG) was then used to study the effect of complete [Ca2+](er) emptying during the oxidative stress generated by Asc/Men. Surprisingly, the combination of TG and Asc/Men increased ER stress to a level considerably higher than that observed for either treatment alone, suggesting that [Ca2+](er) release alone is not sufficient to explain ER stress activation during oxidative stress. Finally, TG-mediated [Ca2+](er) release largely potentiated ER stress, DNA fragmentation and cell death caused by Asc/Men, supporting a role of ER stress in the process of Asc/Men cytotoxicity. Taken together, our results highlight the involvement of ER stress and [Ca2+](er) decrease in the process of oxidative stress-induced cell death in MCF-7 cells. PMID- 20006590 TI - Beta tubulin affects the aryl hydrocarbon receptor function via an Arnt-mediated mechanism. AB - We have been studying the requirement for the aryl hydrocarbon receptor nuclear translocator (Arnt)-dependent DNA complex formation, which precedes the activation of gene transcription. Using DEAE chromatography, we have obtained a Sf9 insect fraction F5 that is highly enriched with beta-tubulin. F5 inhibits the formation of the AhR gel shift complex and this inhibition is sensitive to protease, suggesting that proteins that are present in this F5 fraction are responsible for the inhibition. Additional experiments have revealed that this inhibition is less pronounced in the presence of anti-beta-tubulin IgG and beta tubulin enriched fraction from pig brain also inhibits the AhR gel shift formation. Sf9 beta-tubulin interacts with Arnt and suppresses the binding of the AhR/Arnt heterodimer to its corresponding enhancer. Human beta4-tubulin, which shares high sequence identity with Sf9 beta-tubulin, suppresses the AhR-dependent luciferase expression by reducing the nuclear Arnt content and retaining Arnt in the cytoplasm. Fluorescence studies using the GFP fusion of human beta4-tubulin have revealed that beta4-tubulin prevents the localization of Arnt in Sf9 cells. Here we have provided evidence suggesting that beta-tubulin may regulate the physiological content of Arnt. PMID- 20006591 TI - The isothiocyanate produced from glucomoringin inhibits NF-kB and reduces myeloma growth in nude mice in vivo. AB - Glucosinolates (GLs), natural compounds extracted from Brassicaceae and precursors of isothiocyanates (ITCs), have been studied in the last decades mostly due to their chemopreventive activity and, more recently, for their potential use as novel chemotherapeutics. The aim of the present study was to investigate the in vitro and in vivo activity of glucomoringin (GMG), an uncommon member of the GLs family, and to compare it with glucoraphanin (GRA), one of the most studied GL. We have evaluated the potency of both compounds in inducing cell death, cell cycle perturbations, apoptosis, NF-kB inhibition and GST-pi activity in human carcinoma cells with different GST-pi contents as well as in human multiple myeloma and leukaemia cell lines. GMG-derived ITC (GMG-ITC) showed to be more effective compared to GRA-derived ITC (Sulforaphane), especially in inhibiting NF-kB activity and inducing apoptosis through a caspase-dependent pathway; these effects were more pronounced in myeloma cells, in which we could also observe a long lasting growth inhibitory effect, probably due to NF-kB inhibition, which is considered essential for myeloma cell survival. Both GLs were able to induce cell death in the muM range in all tested cell lines but caused cell cycle perturbations only in myeloma cells; they were also able to modulate the GST/GSH pathway by causing a 3-fold increase in GST-pi activity in MCF7 cells. In vivo study showed that pure GMG-ITC was only slightly active in a carcinoma mice model, whereas it had significant antitumoral activity in a myeloma model, causing little toxicity. PMID- 20006592 TI - Light up and see: enhancement of the visual mismatch negativity (vMMN) by nicotine. AB - Both smoking and nicotine can facilitate cognitive efficiency in humans, however the exact mechanism underlying this improvement in cognitive performance is unclear. Nicotine-related improvements in visual task performance may stem from facilitation of the identification and encoding of rare deviant stimuli at early sensory levels. Visual processes at these early levels are thought to be indexed by the visual mismatch negativity (vMMN), an event-related potential (ERP) measure of pre-conscious deviant detection. In order to contribute to our understanding of the neural mechanisms underlying nicotinic modulated cognition, the current study investigated the acute effects of nicotine on vMMN in a non smoking sample. Twenty-seven volunteers (7 males, 20 females) were treated with nicotine gum (6 mg) in a double-blind randomized, placebo-controlled repeated measures design. ERPs (vMMN; visual N100 and P200) and motor indices of performance were extracted from an intermodal task, requiring participants to attend selectively to auditory targets presented within concurrent, non overlapping oddball sequences of visual standard and deviant stimuli. Behavioural performance was unaffected by nicotine, however nicotine was found to enhance vMMN and P200 amplitude. The findings are discussed in relation to attentional and neurobiological theories of nicotine dependence and of cognition in general. PMID- 20006593 TI - Prevalence of human anti-mouse antibodies (HAMAs) in routine examinations. AB - BACKGROUND: Circulating heterophilic antibodies interfere with immunological assays in laboratory examinations; however, their rate of incidence is currently questionable. We developed an enzyme-linked immunosorbent assay (ELISA) to detect human anti-mouse antibodies (HAMAs) in routine examinations. METHODS: The study samples were comprised of serum samples obtained from 290 inpatients and outpatients at our hospital. Mouse immunoglobulin G1 (mIgG1), mIgG2a, and mIgG2b were used as the antigens and horseradish peroxidase (HRP)-conjugated anti-human IgG and IgM were used to identify the HAMA isotype. RESULTS: HAMAs were detected in 11.7% (34/290) of the samples. We observed 18 and 20 samples positive for IgG- and IgM-type HAMAs, respectively. Four samples contained both IgG- and IgM-type HAMAs. HAMAs against mIgG1, mIgG2a, and mIgG2b were found in 21, 14, and 13 samples, respectively. Existence of HAMAs was confirmed by western blotting using mIgG's as the antigens and HAMAs as the primary antibodies. Heterophilic blocking reagent (HBR) was also used to block the heterophilic interactions. Unexpectedly, a low HBR concentration rather enhanced the interactions instead of blocking them. CONCLUSIONS: A considerable number of HAMA-positive samples, reacting with the heavy chain of mIg, were found in routine examinations. A sufficient amount of HBR should be used for blocking the heterophilic interactions. PMID- 20006594 TI - FcRn binding properties of an abnormal truncated analbuminemic albumin variant. AB - BACKGROUND: : The major histocompatibility class I-related neonatal Fc receptor, FcRn, salvages both IgG and albumin from degradation and thus contributes to maintain high serum levels of these proteins. Analbuminemia is a rare autosomal recessive disorder characterized by clinically observed allelic albumin variants that are absent or found in very low concentrations in the blood circulation. Such variants may have altered FcRn binding properties that affect their half life, biodistribution and thereby transport ability. METHODS: : We established an easy cloning, expression and purification strategy to obtain recombinant GST tagged human serum albumin (HSA) variants for evaluation of pH dependent FcRn binding properties using an enzyme-linked immunosorbent assay (ELISA) and a real time surface plasmon resonance (SPR) biosensor system. RESULTS: : The strategy yielded purified GST-tagged albumin variants. A recombinant truncated HSA variant similar to a clinically observed splice mutant denoted Bartin, here abrogated HSA(Bartin), showed no detectable pH dependent FcRn binding compared to a fully functional albumin wild type variant, HSA(Wt), and a truncated HSA variant consisting of only the carboxy terminal domain III (HSA(DIII)). CONCLUSIONS: : The approach described can be used to rapidly screen clinically observed truncated or otherwise mutant or modified HSA variants regarding their pH dependent FcRn binding properties. Here, we demonstrate that a recombinant truncated HSA variant, HSA(Bartin), does not interact with FcRn, which gives a molecular explanation for the low serum levels. In addition, DIII of HSA alone was shown to retain its FcRn binding property. PMID- 20006595 TI - Activation and expression of urokinase-type plasminogen activator are modulated by freezing/thawing process through activation of redox signal pathway in primary porcine endometrial cells. AB - Plasminogen activators (PAs) play a pivotal role in a variety of uterine physiologies, such as endometrial function, trophoblast invasion, and implantation process, but its alteration in expression or activity during cryopreservation of primary uterine cells has received little attention. In this study, we investigated whether PA expression and activity were modulated in first passage primary porcine uterus endometrial epithelium cells (PUEECs) treated with or without a freezing-thawing procedure. Western blotting and zymographic analysis showed that uPA expression and activity increased significantly in frozen-thawed PUEECs in a passage-dependent manner as compared to freshly prepared control cells. Moreover, intracellular reactive oxygen species (ROS) were increased by freezing-thawing and longer culturing, and were more prominent in frozen-thawed PUEECs than in control cells. However, the increase in both uPA expression and activity was greatly reduced or alleviated by treatment with either ROS scavenger N-acetylcysteine or extracellular signal-regulated kinase (ERK) inhibitor PD98059. These results suggest that ROS/ERK-mediated uPA activation may be an important factor in cryo-damage of primary uterine cells. PMID- 20006597 TI - Antagonists and substrates differentially regulate serotonin transporter cell surface expression in serotonergic neurons. AB - The serotonin transporter (SERT) terminates serotonergic neurotransmission by the rapid removal of serotonin (5-hydroxytryptamine, 5-HT) from the extracellular space back into serotonergic neurons. SERT therefore controls the concentration of extracellular 5-HT, and thus one mechanism to regulate the efficacy of serotonergic neurotransmission is via modulation of the density of SERT molecules on the cell membrane. We have studied the effects of prolonged exposure to various selective serotonin re-uptake inhibitors (SSRIs), as well as cocaine and the transport substrates 5-HT and 3,4-methylenedioxy-methamphetamine (MDMA), on SERT cell surface expression in cultured serotonergic neurons. This was achieved via quantification of the amount of cell surface-expressed SERT molecules using antibody detection combined with confocal laser scanning microscopy. Our results show that exposure to the SSRIs citalopram, fluoxetine, sertraline and paroxetine all induced SERT internalization, but with different efficacies. The substrates 5 HT and MDMA also induced SERT internalization, while cocaine elevated SERT cell surface expression. PMID- 20006598 TI - Attenuation of experimental autoimmune encephalomyelitis in C57 BL/6 mice by osthole, a natural coumarin. AB - Osthole, a natural coumarin, is known to have a variety of pharmacological and biochemical uses and is considered to have potential therapeutic applications. Here we examined the effects of osthole on the central nervous system demyelination in experimental autoimmune encephalomyelitis (EAE), a model of multiple sclerosis and its mechanism(s). C57 BL/6 mice immunized with myelin oligodendrocyte glycoprotein 35-55 amino acid peptide were treated with osthole at day 7 post immunization (7 p.i., subclinical periods, early osthole treatment) and day 13 p.i. (clinical periods, late osthole treatment) respectively and both therapies continued throughout the study. The content of nerve growth factor (NGF) and interferon gamma (IFN-gamma) in the sera and brain of mice in vivo as well as the splenocytes culture supernatants in vitro were detected. The results showed that osthole retarded the disease process when the therapy was initiated at subclinical periods, attenuated the clinical severity of EAE mice when the therapy was initiated at both subclinical and clinical periods, ameliorated inflammation and demyelination and improved the outcomes of magnetic resonance imaging. In addition, osthole blocked the reduction of NGF and suppressed IFN gamma increase in EAE mice. These results suggested that osthole might be a new pharmacological approach to treat multiple sclerosis. PMID- 20006596 TI - Pharmacological profile of the NOP agonist and cough suppressing agent SCH 486757 (8-[Bis(2-Chlorophenyl)Methyl]-3-(2-Pyrimidinyl)-8-Azabicyclo[3.2.1]Octan-3-Ol) in preclinical models. AB - We describe the pharmacological and pharmacokinetic profiles of SCH 486757, a nociceptin/orphanin FQ peptide (NOP) receptor agonist that has recently entered human clinical trials for cough. SCH 486757 selectively binds human NOP receptor (K(i)=4.6+/-0.61nM) over classical opioid receptors. In a guinea pig capsaicin cough model, SCH 486757 (0.01-1mg/kg) suppressed cough at 2, 4, and 6h post oral administration with a maximum efficacy occurring at 4h equivalent to codeine, hydrocodone, dextromethorphan and baclofen. The antitussive effects of SCH 486757 (3.0mg/kg, p.o.) was blocked by the NOP receptor antagonist J113397 (12mg/kg, i.p.) but not by naltrexone (10mg/kg, p.o.). SCH 486757 does not produce tolerance to its antitussive activity after a 5-day BID dosing regimen. After acute and chronic dosing paradigms, SCH 486757 (1mg/kg) inhibited capsaicin evoked coughing by 46+/-9% and 40+/-11%, respectively. In a feline mechanically evoked cough model, SCH 486757 produces a maximum inhibition of cough and expiratory abdominal electromyogram amplitude of 59 and 61%, respectively. SCH 486757 did not significantly affect inspiratory electromyogram amplitude. We examined the abuse potential of SCH 486757 (10mg/kg, p.o.) in a rat conditioned place preference procedure which is sensitive to classical drugs of abuse, such as amphetamine and morphine. SCH 486757 was without effect in this model. Finally, SCH 486757 displays a good oral pharmacokinetic profile in the guinea pig, rat and dog. We conclude that SCH 486757 has a favorable antitussive profile in preclinical animal models. PMID- 20006599 TI - 4-Hydroxytamoxifen inhibits K(+) currents in mouse ventricular myocytes. AB - Tamoxifen is a widely used chemotherapeutic agent, which has been associated with prolongation of the QT interval. Other studies have reported that acute exposure to tamoxifen can reduce cardiac K(+) currents. However, in vivo tamoxifen is largely metabolized and most of its activity is attributable to its major metabolite, 4-hydroxytamoxifen (4OH-tamoxifen). Accordingly, in the present study, we performed voltage-clamp experiments to directly investigate the effects of 4OH-tamoxifen on the repolarizing K(+) currents in adult mouse ventricular myocytes in order to determine whether the effects of tamoxifen on repolarization could be ascribed to 4OH-tamoxifen. K(+) currents were recorded before and after acute exposure to 4OH-tamoxifen (0.5, 1 and 10microM). 4OH-tamoxifen reduced the density of the Ca(2+)-independent transient outward (I(to)), the ultrarapid delayed rectifier (I(Kur)) and the inward rectifier (I(K1)) K(+) currents (by up to 43%, 41% and 26%, respectively) but had no significant effect on the steady state outward K(+) current (I(ss)). Voltage dependence of steady-state inactivation and reactivation time of I(to) and I(Kur) were not affected by 4OH tamoxifen. Experiments using the pure estrogen receptor antagonist, ICI 182,780 and the inhibitor of gene transcription, actinomycin D, were undertaken to assess the involvement of estrogen receptor. Administered alone these compounds did not affect the density of K(+) currents. Moreover, pretreatment of the cells with ICI 182,780 or actinomycin D did not prevent the inhibitory response to 4OH tamoxifen. Overall, 4OH-tamoxifen reduced K(+) currents in mouse ventricle and this effect is unrelated to gene transcription and does not involve interaction of the drug with estrogen receptor. PMID- 20006600 TI - Differential effects of systemic and intraseptal administration of the acetylcholinesterase inhibitor tacrine on the recovery of spatial behavior in an animal model of diencephalic amnesia. AB - Several lines of evidence suggest that acetylcholinesterase inhibitors (AChE) have their cognitive enhancing effects by stimulating cholinergic receptors within the medial septum. However, intraseptal administration of cholinergic enhancing drugs produce mixed results that appear to depend on both the integrity of the medial septum as well as task demands. Three experiments were conducted to determine the relationship between increased cholinergic activity within the medial septum and hippocampus and behavioral recovery in a model of diencephalic amnesia produced by pyrithiamine-induced thiamine deficiency (PTD). In Experiment 1, systemic tacrine (0.0, 0.75, 1.5mg/kg) was administered to PTD and pair-fed (PF) rats prior to a spontaneous alternation task. Without tacrine, PF rats alternated at a higher rate than PTD rats. Both doses of tacrine increased alternation in PTD rats to within the range of PF rats. In Experiment 2, three doses of intraseptal tacrine (2.5, 5.0, 12.5microg) were administered to PTD and PF rats and changes in hippocampal acetylcholine efflux were assessed. Both the 5.0 and 12.5microg doses significantly increased hippocampal acetylcholine levels, but the change was greater in the PTD rats. In Experiment 3, despite the fact that both intraseptal doses of tacrine (5.0, 12.5microg) increased hippocampal acetylcholine levels, only 5.0microg significantly improved alternation scores in PTD rats. Thus, when there is basal forebrain cholinergic cell loss in conjunction with diencephalic pathology, the therapeutic range of AChE-I in the medial septum and the effective doses do not directly map onto changes in acetylcholine efflux in the hippocampus. PMID- 20006601 TI - ADAMTS-1 contributes to the antifibrotic effect of captopril by accelerating the degradation of type I collagen in chronic viral myocarditis. AB - Angiotensin-converting enzyme (ACE) inhibitors have been demonstrated to have antifibrotic activity in myocardial fibrosis. A distintegrin and metalloprotease with thrombospondin type 1 motifs (ADAMTS-1) is a newly discovered metalloproteinase. It was reported ADAMTS-1 had novel gelatin (type I collagen) degrading activities. We examined the role of ADAMTS-1 in the antifibrotic activity of the ACE inhibitor Captopril in a chronic viral myocarditis (CVMC) model. Balb/c mice were assigned to five groups: normal control group1 (group 1), normal control group2 (group 2), CVMC model group (group 3), CVMC control group (group 4) and Captopril therapy group (group 5). Group 3, 4 and 5 received Coxsackievirus B(3) to induce CVMC and group 5 was treated with Captopril (100mg/kg)for 28days. Heart sections were stained with picrosirius red and collagen volume fraction calculated. ADAMTS-1 expression was determined by Western blot. Type I collagen and carboxyterminal telopeptide of type I collagen (ICTP) were measured by RT-PCR. Group 4 mice had significantly increased collagen volume fraction compared to groups 2 and 5 (P<0.001, P<0.001, respectively) and higher type I collagen mRNA expression than groups 2 and 5 (P<0.001, P<0.001, respectively). Group 5 ADAMTS-1 and ICTP expression was significantly higher than in groups 2 and 4 (P<0.001, P<0.001, respectively). ADAMTS-1 levels in group 5 negatively correlated with collagen volume fraction (r=-0.68, P<0.01) and type I collagen (r=-0.67, P<0.01) but positively correlated with ICTP (r=0.72, P<0.01). We conclude that ADAMTS-1 contributes to the antifibrotic effect of Captopril by accelerating the degradation of type I collagen in CVMC. PMID- 20006602 TI - Anti-inflammatory and immunosuppressive effects of the enaminone E121. AB - Asthma is a chronic inflammatory disease of the airways. The treatment of asthma is far from optimal and hence the need for novel therapeutic agents exists. The purpose of this study was to assess the anti-asthma effects of an enaminone, E121, and also its effects on human peripheral blood mononuclear cell proliferation and cytokine release. The effects of E121 were assessed in an ovalbumin-induced model of airway inflammation and airway hyperresponsiveness. In addition, the effects of E121 on phytohemagglutinin (PHA), anti-CD3 monoclonal antibody and lipopolysaccharide (LPS)-induced human peripheral blood mononuclear cell proliferation and cytokine release, respectively, were assessed. Treatment of mice with E121 significantly decreased the ovalbumin-induced increase in airway total cell influx and eosinophil infiltration and this was associated with an inhibition of ovalbumin-induced airway hyperresponsiveness. Moreover, E121 reduced PHA and anti-CD3-induced human peripheral blood mononuclear cell proliferation in vitro. E121 also inhibited PHA, anti-CD3 monoclonal antibody and LPS-induced cytokine release from human peripheral blood mononuclear cell cultures. These findings indicate that E121 exhibits anti-inflammatory and immunosuppressive activities. PMID- 20006603 TI - Osteoblasts extracellular matrix induces vessel like structures through glycosylated collagen I. AB - Extracellular matrix (ECM) plays a fundamental role in angiogenesis affecting endothelial cells proliferation, migration and differentiation. Vessels-like network formation in vitro is a reliable test to study the inductive effects of ECM on angiogenesis. Here we utilized matrix deposed by osteoblasts as substrate where the molecular and structural complexity of the endogenous ECM is preserved, to test if it induces vessel-like network formation by endothelial cells in vitro. ECM is more similar to the physiological substrate in vivo than other substrates previously utilized for these studies in vitro. Osteogenic ECM, prepared in vitro from mature osteoblasts at the phase of maximal deposition and glycosylation of collagen I, induces EAhy926, HUVEC, and HDMEC endothelial cells to form vessels-like structures and promotes the activation of metalloproteinase 2 (MMP-2); the functionality of the p-38/MAPK signaling pathway is required. Osteogenic ECM also induces a transient increase of CXCL12 and a decrease of the receptor CXCR4. The induction of vessel-like networks is dependent from proper glycosylation of collagens and does not occur on osteogenic ECMs if deglycosylated by -galactosidase or on less glycosylated ECMs derived from preosteoblasts and normal fibroblasts, while is sustained on ECM from osteogenesis imperfecta fibroblasts only when their mutation is associated with over-glycosylation of collagen type I. These data support that post-translational glycosylation has a role in the induction in endothelial cells in vitro of molecules conductive to self-organization in vessels-like structures. PMID- 20006604 TI - Non-cell-autonomous stimulation of stem cell proliferation following ablation of Tcf3. AB - A combination of cell intrinsic factors and extracellular signals determine whether mouse embryonic stem cells (ESC) divide, self-renew, and differentiate. Here, we report a new interaction between cell intrinsic aspects of the canonical Wnt/Tcf/beta-catenin signaling pathway and extracellular Lif/Jak/Stat3 stimulation that combines to promote self-renewal and proliferation of ESC. Mutant ESC lacking the Tcf3 transcriptional repressor continue to self-renew in the absence of exogenous Lif and through pharmacological inhibition of Lif/Jak/Stat3 signaling; however, proliferation rates of TCF3-/- ESC were significantly decreased by inhibiting Jak/Stat3 activity. Cell mixing experiments showed that stimulation of Stat3 phosphorylation in TCF3-/- ESC was mediated through secretion of paracrine acting factors, but did not involve elevated Lif or LifR transcription. The new interaction between Wnt and Lif/Jak/Stat3 signaling pathways has potential for new insights into the growth of tumors caused by aberrant activity of Wnt/Tcf/beta-catenin signaling. PMID- 20006605 TI - The human IFN-inducible p53 target gene TRIM22 colocalizes with the centrosome independently of cell cycle phase. AB - TRIM22 (Staf50), a member of the TRIM protein family, is an interferon (IFN) inducible protein as well as a p53 target gene. The function of TRIM22 is largely unknown, but TRIM22 is suggested to play a role in viral defense by restriction of viral replication. In addition, TRIM22 may function as a ubiquitin E3 ligase. In contrast to previous reports showing solely cytoplasmic localization of exogenous TRIM22, we report here that endogenous TRIM22 is localized to both nucleus and cytosol in primary human mononuclear cells, as well as in the human osteosarcoma cell line U2OS. Moreover, we demonstrate a colocalization of TRIM22 with the centrosomes in primary cells as well as in U2OS cells, and show that this colocalization is independent of cell cycle phase. Additionally, our data suggest the colocalization with centrosomes to be independent on the microtubule network. Given that some viral protein assembly takes place in the close vicinity of the centrosome, our data suggest that important functions of TRIM22 such as regulation of viral replication and protein degradation may take place in the centrosome. However, further studies are warranted to certify this notion. PMID- 20006606 TI - VEGF elicits epithelial-mesenchymal transition (EMT) in prostate intraepithelial neoplasia (PIN)-like cells via an autocrine loop. AB - Vascular endothelial growth factor (VEGF) is overexpressed during the transition from prostate intraepithelial neoplasia (PIN) to invasive carcinoma. We have mimicked such a process in vitro using the PIN-like C3(1)/Tag-derived Pr-111 cell line, which expresses low levels of VEGF and exhibits very low tumorigenicity in vivo. Elevated expression of VEGF164 in Pr-111 cells led to a significant increase in tumorigenicity, invasiveness, proliferation rates and angiogenesis. Moreover, VEGF164 induced strong changes in cell morphology and cell transcriptome through an autocrine mechanism, with changes in TGF-beta1- and cytoskeleton-related pathways, among others. Further analysis of VEGF overexpressing Pr-111 cells or following exogenous addition of recombinant VEGF shows acquisition of epithelial-mesenchymal transition (EMT) features, with an increased expression of mesenchymal markers, such as N-cadherin, Snail1, Snail2 (Slug) and vimentin, and a decrease in E-cadherin. Administration of VEGF led to changes in TGF-beta1 signaling, including reduction of Smad7 (TGF-beta inhibitory Smad), increase in TGF-betaR-II, and translocation of phospho-Smad3 to the nucleus. Our results suggest that increased expression of VEGF in malignant cells during the transition from PIN to invasive carcinoma leads to EMT through an autocrine loop, which would promote tumor cell invasion and motility. Therapeutic blockade of VEGF/TGF-beta1 in PIN lesions might impair not only tumor angiogenesis, but also the early dissemination of malignant cells outside the epithelial layer. PMID- 20006607 TI - The omega-3 fatty acid, DHA, decreases neuronal cell death in association with altered zinc transport. AB - Docosahexaenoic acid (DHA) is the major polyunsaturated fatty acid in neuronal cell membranes. We hypothesize that DHA induces a decrease in neuronal cell death through reduced ZnT3 expression and zinc uptake. Exposure of M17 cells to DHA deficient medium increased the levels of active caspase-3, relative to levels in DHA-replete cells, confirming the adverse effects of DHA deficiency in promoting neuronal cell death. In DHA-treated M17 cells, zinc uptake was 65% less and ZnT3 mRNA and protein levels were reduced in comparison with DHA-depleted cells. We propose that the neuroprotective function of DHA is exerted through a reduction in cellular zinc levels that in turn inhibits apoptosis. PMID- 20006608 TI - Sphingolipids and gangliosides of the nervous system in membrane function and dysfunction. AB - Simple sphingolipids such as ceramide and sphingomyelin (SM) as well as more complex glycosphingolipids play very important roles in cell function under physiological conditions and during disease development and progression. Sphingolipids are particularly abundant in the nervous system. Due to their amphiphilic nature they localize to cellular membranes and many of their roles in health and disease result from membrane reorganization and from lipid interaction with proteins within cellular membranes. In this review we discuss some of the functions of sphingolipids in processes that entail cellular membranes and their role in neurodegenerative diseases, with an emphasis on SM, ceramide and gangliosides. PMID- 20006609 TI - Sharp-1 modulates the cellular response to DNA damage. AB - DNA damage checkpoints are essential for maintenance of genome integrity. We report here that inducible overexpression of the transcription factor Sharp-1 results in an S and G2/M cell cycle arrest, concomitant with the upregulation of Brca1 and GADD45alpha expression. In addition, we show that endogenous Sharp-1 mRNA is increased by DNA-damaging agents. Consistently, Sharp-1 overexpressing cells exhibit reduced apoptosis in response to chemotherapeutic drugs along with lower p53 expression and activity. Our studies identify a novel function for Sharp-1 in cell cycle arrest and DNA damage-induced apoptosis. Inappropriate Sharp-1 expression may therefore be associated with tumorigenesis. PMID- 20006610 TI - Disease-associated variants of microsomal retinol dehydrogenase 12 (RDH12) are degraded at mutant-specific rates. AB - Mutations in retinol dehydrogenase 12 (RDH12) cause severe retinal degeneration. However, some of the disease-associated RDH12 mutants retain significant catalytic activity, indicating the existence of additional pathophysiological mechanisms. This study demonstrates that the catalytically active T49M and I51N mutants undergo accelerated degradation, which results in their reduced cellular levels. Inhibition of proteasome leads to significant accumulation of ubiquitylated T49M and I51N. Furthermore, the degree of ubiquitylation strongly correlates with the half-lives of the proteins. These results suggest that the accelerated degradation of RDH12 mutants by the ubiquitin-proteasome system contributes to the pathophysiology and phenotypic variability associated with mutations in the RDH12 gene. PMID- 20006611 TI - Glyceraldehyde-3-phosphate dehydrogenase tetramer dissociation and amyloid fibril formation induced by negatively charged membranes. AB - Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) is a multifunctional enzyme related with Huntington's, Parkinson's and Alzheimer's diseases. The ability of negatively charged membranes to induce a rapid formation of GAPDH amyloid fibrils has been demonstrated, but the mechanisms by which GAPDH reaches the fibrillar state remains unclear. In this report, we describe the structural changes undergone by GAPDH at physiological pH and temperature conditions right from its interaction with acidic membranes until the amyloid fibril is formed. According to our results, the GAPDH-membrane binding induces a beta-structuring process along with a loss of quaternary structure in the enzyme. In this way, experimental evidences on the initial steps of GAPDH amyloid fibrils formation pathway are provided. PMID- 20006612 TI - Resistance to direct antiviral agents in patients with hepatitis C virus infection. AB - Chronic hepatitis C virus (HCV) infection is one of the major causes of cirrhosis, hepatocellular carcinoma, and liver failure that leads to transplantation. The current standard treatment, a combination of pegylated interferon alfa and ribavirin, eradicates the virus in only about 50% of patients. Directly acting antiviral (DAA) agents, which inhibit HCV replication, are in phase 1, 2, and 3 trials; these include reagents that target the nonstructural (NS)3 protease, the NS5A protein, the RNA-dependent RNA-polymerase NS5B, as well as compounds that directly inhibit HCV replication through interaction with host cell proteins. Because of the high genetic heterogeneity of HCV and its rapid replication, monotherapy with DAA agents poses a high risk for selection of resistant variants. We review the parameters that determine resistance, genotypic and phenotypic resistance profiles of DAA agents, and strategies to avoid the selection of resistant variants. PMID- 20006613 TI - Selenium status and the risk of esophageal and gastric cancer subtypes: the Netherlands cohort study. AB - BACKGROUND & AIMS: Selenium may protect against the development of esophageal squamous cell carcinoma (ESCC), esophageal adenocarcinoma (EAC), and gastric cardia adenocarcinoma (GCA). Only in very few studies have the associations with ESCC and GCA been investigated, and no epidemiologic studies exist on EAC. METHODS: We studied the association between selenium and risk of ESCC, EAC, and GCA within the prospective Netherlands Cohort Study, conducted among 120,852 men and women aged 55-69 years at baseline. In September 1986, the cohort members completed a questionnaire on risk factors for cancer and provided toenail clippings for determination of baseline selenium status. After 16.3 years of follow-up, 64 ESCC, 112 EAC, and 114 GCA cases and 2072 subcohort members were available for case-cohort analysis. Incidence rate ratios (RR) were calculated using Cox proportional hazards models. RESULTS: In multivariable analyses of selenium status, we found an inverse association with ESCC (RR(per standard unit increment), 0.80; 95% confidence interval [CI]: 0.67-0.96) and a borderline significant inverse association with GCA (RR, 0.91; 95% CI: 0.80-1.02). No overall association was observed for EAC (RR, 1.05; 95% CI: 0.95-1.15), but, for women and never smokers, significant inverse associations were found (RR(per standard unit increment), 0.72; 95% CI: 0.61-0.84 and RR(per standard unit increment), 0.74; 95% CI: 0.64-0.86, respectively). CONCLUSIONS: This prospective study supports an inverse association between toenail selenium and risk of ESCC and GCA and suggests an inverse association with risk of EAC in subgroups (women, never smokers, and low antioxidant consumers). These associations need confirmation. PMID- 20006614 TI - Do social disadvantage and early family adversity affect the diurnal cortisol rhythm in infants? The Generation R Study. AB - Dysregulation of diurnal cortisol secretion patterns may explain the link between adversities early in life and later mental health problems. However, few studies have investigated the influence of social disadvantage and family adversity on the hypothalamic-pituitary-adrenal (HPA) axis early in life. In 366 infants aged 12-20 months from the Generation R Study, a population-based cohort from fetal life onwards, parents collected saliva samples from their infant at 5 moments over the course of 1 day. The area under the curve (AUC), the cortisol awakening response (CAR) and the diurnal cortisol slope were calculated as different composite measures of the diurnal cortisol rhythm. Information about social disadvantage and early adversity was collected using prenatal and postnatal questionnaires. We found that older infants showed lower AUC levels; moreover, infants with a positive CAR were significantly older. Both the AUC and the CAR were related to indicators of social disadvantage and early adversity. Infants of low income families, in comparison to high income families, showed higher AUC levels and a positive CAR. Infants of mothers who smoked during pregnancy were also significantly more likely to show a positive CAR. Furthermore, infants of mothers experiencing parenting stress showed higher AUC levels. The results of our study show that effects of social disadvantage and early adversity on the diurnal cortisol rhythm are already observable in infants. This may reflect the influence of early negative life events on early maturation of the HPA axis. PMID- 20006615 TI - Small interfering RNA (siRNA) delivery into murine bone marrow-derived macrophages by electroporation. AB - Selective gene silencing by RNA interference (RNAi) is a valuable tool for the targeted manipulation of the development and/or function of cells. Using a fluorescein-labeled non-silencing siRNA duplex, we established a protocol for the electroporation of primary mouse macrophages which routinely yielded >95% transfected cells. Electroporation of siRNAs directed against MAPK1 and CD86 led to an efficient knock-down of cellular protein in bone marrow-derived mouse macrophages (BM-Mphi). Importantly, the electroporation procedure did not impair the viability of BM-Mphi, their ability to ingest or degrade E. coli or their capacity to express iNOS mRNA, to produce NO or to upregulate TNF and IL-6 mRNA in response to inflammatory stimuli such as LPS. Therefore, we propose that electroporation of silencing siRNAs into murine BM-Mphi is a highly efficient method to manipulate gene expression of BM-Mphi that does not cause toxicity or a non-specific alteration of macrophage biology. PMID- 20006616 TI - Formulation of immunoassay calibrators in pasteurized albumin can significantly enhance their durability. AB - Calibrator matrix can have significant effects on the commutability of assay standards and on the maintenance of their integrity. We have observed marked instability in progastrin-releasing peptide (proGRP) assay standards traceable to the bovine serum albumin (BSA) used in matrix formulation. Attempts were made to improve calibrator stability using different albumin pretreatments. Observed analyte recoveries in calibrators prepared with untreated BSA were consistently less than 45% after 1 week of storage at 4 degrees C. Pre-treating the BSA by chromatography on immobilized heparin or benzamidine failed to improve calibrator durability with day 7 recoveries of less than 55%. In marked contrast, calibrators formulated with albumin pasteurized at pH 3.0 displayed remarkable stability. Recoveries of >97% were observed after 4 weeks of storage at either 4 degrees C or room temperature. Even calibrators incubated for 4 weeks at 37 degrees C gave recoveries between 91-106%. This improvement was not seen with BSA pasteurized at neutral pH. Albumin pretreatment is straightforward, easily scalable and dramatically improves calibrator stability. Matrix formulated with acid-pasteurized BSA may prove more generally useful when assays are plagued by poor calibrator durability. PMID- 20006617 TI - Venom of the egg-larval parasitoid Chelonus inanitus is a complex mixture and has multiple biological effects. AB - The egg-larval parasitoid Chelonus inanitus injects bracoviruses (BVs) and venom along with the egg into the host egg; both components are essential for successful parasitoid development. All stages of eggs of its natural host, Spodoptera littoralis, can be successfully parasitized, i.e. from mainly a yolk sphere to a fully developed embryo. Here, we show that the venom contains at least 25 proteins with masses from 14kDa to over 300kDa ranging from acidic to basic. The majority is glycosylated and their persistence in the host is short when old eggs are parasitized and much longer when young eggs are parasitized. Physiological experiments indicated three different functions. (1) Venom synergized the effect of BVs in disrupting host development when injected into third instar larvae. (2) Venom had a transient paralytic effect when injected into sixth instar larvae. (3) In vitro experiments with haemocytes of fourth instar larvae suggested that venom alters cell membrane permeability. We propose that venom promotes entry of BVs into host cells and facilitates placement of the egg in the embryo's haemocoel when old eggs are parasitized. The multifunctionality of the venom might thus be essential in enabling parasitization of all stages of host eggs. PMID- 20006618 TI - Molecular determinants required for selective interactions between the thyroid hormone receptor homodimer and the nuclear receptor corepressor N-CoR. AB - The unliganded nuclear receptor (NR) generally recruits the NR corepressor (N CoR) and the silencing mediator of retinoid and thyroid hormone receptor via its direct binding to the extended helical motif within dual NR-interaction domains (IDs) of corepressors. Interestingly, N-CoR has a third ID (ID3) upstream of two IDs (ID1 and ID2) and its core motif (IDVII), rather than an extended helical motif, is known to be involved directly in the exclusive interaction of ID3 with the thyroid hormone receptor (TR). Here, we investigated the molecular determinants of the TR interaction with ID3 to understand the molecular basis of the N-CoR preference shown by the TR homodimer. Using a one- plus two-hybrid system, we identified the specific residues of N-CoR-ID2 and N-CoR-ID3 that are required for stable association of N-CoR with the TR homodimer. By swapping experiments and mutagenesis studies, we found that the C-terminally flanked residues of the core motif of ID3 contribute to the TR preference for N-CoR-ID3, suggesting that an extended three-turn helix might form within the ID3 via a C terminal extension (IDVIITRQI) and participate directly in the TR-specific interaction. Structural modeling of the ID3 motif on TR-LBD is consistent with this conclusion. Notably, we identified a novel interaction between N-CoR-ID3 and orphan NR RevErb that is mediated by the residues crucial also in TR binding. These observations raise the intriguing possibility that NR homodimers such as TR and RevErb display preferential binding to the N-CoR corepressor via their specific interactions with ID3, which is normally absent from the silencing mediator of retinoid and thyroid hormone receptor. PMID- 20006619 TI - Characterisation of a GroEL single-ring mutant that supports growth of Escherichia coli and has GroES-dependent ATPase activity. AB - Binding and folding of substrate proteins by the molecular chaperone GroEL alternates between its two seven-membered rings in an ATP-regulated manner. The association of ATP and GroES to a polypeptide-bound ring of GroEL encapsulates the folding proteins in the central cavity of that ring (cis ring) and allows it to fold in a protected environment where the risk of aggregation is reduced. ATP hydrolysis in the cis ring changes the potentials within the system such that ATP binding to the opposite (trans) ring triggers the release of all ligands from the cis ring of GroEL through a complex network of allosteric communication between the rings. Inter-ring allosteric communication thus appears indispensable for the function of GroEL, and an engineered single-ring version (SR1) cannot substitute for GroEL in vivo. We describe here the isolation and characterisation of an active single-ring form of the GroEL protein (SR-A92T), which has an exceptionally low ATPase activity that is strongly stimulated by the addition of GroES. Dissection of the kinetic pathway of the ATP-induced structural changes in this active single ring can be explained by the fact that the mutation effectively blocks progression through the full allosteric pathway of the GroEL reaction cycle, thus trapping an early allosteric intermediate. Addition of GroES is able to overcome this block by binding this intermediate and pulling the allosteric pathway to completion via mass action, explaining how bacterial cells expressing this protein as their only chaperonin are viable. PMID- 20006620 TI - Crystal structure analysis of free and substrate-bound 6-hydroxy-L-nicotine oxidase from Arthrobacter nicotinovorans. AB - The pathway for oxidative degradation of nicotine in Arthrobacter nicotinovorans includes two genetically and structurally unrelated flavoenzymes, 6-hydroxy-L nicotine oxidase (6HLNO) and 6-hydroxy-D-nicotine oxidase, which act with absolute stereospecificity on the L- and D-forms, respectively, of 6-hydroxy nicotine. We solved the crystal structure of 6HLNO at 1.95 A resolution by combined isomorphous/multiple-wavelength anomalous dispersion phasing. The overall structure of each subunit of the 6HLNO homodimer and the folds of the individual domains are closely similar as in eukaryotic monoamine oxidases. Unexpectedly, a diacylglycerophospholipid molecule was found to be non-covalently bound to each protomer of 6HLNO. The fatty acid chains occupy hydrophobic channels that penetrate deep into the interior of the substrate-binding domain of each subunit. The solvent-exposed glycerophosphate moiety is located at the subunit-subunit interface. We further solved the crystal structure of a complex of dithionite-reduced 6HLNO with the natural substrate 6-hydroxy-L-nicotine at 2.05 A resolution. The location of the substrate in a tight cavity suggests that the binding geometry of this unproductive complex may be closely similar as under oxidizing conditions. The observed orientation of the bound substrate relative to the isoalloxazine ring of the flavin adenine dinucleotide cofactor is suitable for hydride-transfer dehydrogenation at the carbon atom that forms the chiral center of the substrate molecule. A comparison of the substrate-binding modes of 6HLNO and 6-hydroxy-D-nicotine oxidase, based on models of complexes with the D substrate, suggests an explanation for the stereospecificity of both enzymes. The two enzymes are proposed to orient the enantiomeric substrates in mirror symmetry with respect to the plane of the flavin. PMID- 20006621 TI - Discrimination between closely related cellular metabolites by the SAM-I riboswitch. AB - The SAM-I riboswitch is a cis-acting element of genetic control found in bacterial mRNAs that specifically binds S-adenosylmethionine (SAM). We previously determined the 2.9-A X-ray crystal structure of the effector-binding domain of this RNA element, revealing details of RNA-ligand recognition. To improve this structure, variations were made to the RNA sequence to alter lattice contacts, resulting in a 0.5-A improvement in crystallographic resolution and allowing for a more accurate refinement of the crystallographic model. The basis for SAM specificity was addressed by a structural analysis of the RNA complexed to S adenosylhomocysteine (SAH) and sinefungin and by measuring the affinity of SAM and SAH for a series of mutants using isothermal titration calorimetry. These data illustrate the importance of two universally conserved base pairs in the RNA that form electrostatic interactions with the positively charged sulfonium group of SAM, thereby providing a basis for discrimination between SAM and SAH. PMID- 20006622 TI - The prototypical H+/galactose symporter GalP assembles into functional trimers. AB - Glucose is a primary source of energy for human cells. Glucose transporters form specialized membrane channels for the transport of sugars into and out of cells. Galactose permease (GalP) is the closest bacterial homolog of human facilitated glucose transporters. Here, we report the functional reconstitution and 2D crystallization of GalP. Single particle electron microscopy analysis of purified GalP shows that the protein assembles as an oligomer with three distinct densities. Reconstitution assays yield 2D GalP crystals that exhibit a hexagonal array having p3 symmetry. The projection structure of GalP at 18 A resolution shows that the protein is trimeric. Each monomer in the trimer forms its own channel, but an additional cavity (10 approximately 15 A in diameter) is apparent at the 3-fold axis of the oligomer. We show that the crystalline GalP is able to selectively bind substrate, suggesting that the trimeric form is biologically active. PMID- 20006624 TI - Glucose transport from mother to fetus--a theoretical study. AB - The factors that affect and govern the glucose transfer from maternal blood to the fetus are not completely deciphered. We present a steady state, one dimensional mathematical simulation which integrates the main mechanisms that have been shown to exist: metabolic consumption of the placenta, simple and facilitated diffusion via the two membranes of the microvillous and simple diffusion within the placenta. The model uses all available physiologic data we could collect. Numerical results indicate that the most crucial factor in determining the fetal glucose concentration is the facilitated diffusion process at the basal membrane or, more specifically: the permeability of the basal membrane and the density of the transporter GLUT1 on its faces. The gradient between the maternal and the fetal glucose concentration is important as is the metabolic consumption of the placenta. The diffusion within the placenta and the conditions that prevail at the apical microvillous plasma membrane are much less significant. Intrasyncytial concentration of glucose is close to that of maternal blood. The adjustment of the fetal glucose concentration to abrupt changes of its surrounding is estimated to be quite rapid hence for all practical purposes this steady state model can serve as a reasonable approximation. Parameters that await experimental determination are identified. PMID- 20006623 TI - A 2D mechanistic model of breast ductal carcinoma in situ (DCIS) morphology and progression. AB - Ductal carcinoma in situ (DCIS) of the breast is a non-invasive tumor in which cells proliferate abnormally, but remain confined within a duct. Although four distinguishable DCIS morphologies are recognized, the mechanisms that generate these different morphological classes remain unclear, and consequently the prognostic strength of DCIS classification is not strong. To improve the understanding of the relation between morphology and time course, we have developed a 2D in silico particle model of the growth of DCIS within a single breast duct. This model considers mechanical effects such as cellular adhesion and intra-ductal pressure, and biological features including proliferation, apoptosis, necrosis, and cell polarity. Using this model, we find that different regions of parameter space generate distinct morphological subtypes of DCIS, so elucidating the relation between morphology and time course. Furthermore, we find that tumors with similar architectures may in fact be produced through different mechanisms, and we propose future work to further disentangle the mechanisms involved in DCIS progression. PMID- 20006625 TI - Bortezomib suppresses focal adhesion kinase expression via interrupting nuclear factor-kappa B. AB - AIMS: Bortezomib is a potent proteasome inhibitor currently used to treat various malignancies with promising results. To explore the role of bortezomib in reducing cancer cell migration and inducing apoptosis, we evaluated the effects of bortezomib on the expression of focal adhesion kinase (FAK). MAIN METHODS: Various types of cancer cells including lung cancer A549, H1299; a breast cancer MCF7; a hepatocellular carcinoma Huh7, and a tongue squamous cell carcinoma SCC 25 were treated with different concentrations of bortezomib or MG-132 as indicated for 24h. Protein and mRNA levels were determined by Western blotting and real-time PCR. Apoptosis was analyzed by caspase 3 cleavage and activity. FAK promoter and NFkappaB binding activities were measured by luciferase-reporter method. NFkappaB subunit p65 binding capacity was determined by electrophoretic mobility shift assay (EMSA) and chromatin immunoprecipitation (ChIP) analysis. KEY FINDINGS: Both bortezomib and another proteasome inhibitor, MG-132, significantly reduced FAK expression, suppressed cancer cell migration and increased cell apoptosis. Results of real-time PCR and promoter activity assay revealed that bortezomib decreased FAK expression through transcriptional inactivation. Results of FAK promoter activity and ChIP assays in A549 and H1299 cells indicated that bortezomib suppressed FAK activity through a p53-independent pathway. Furthermore, reduction of NFkappaB binding capacity demonstrated by EMSA and ChIP assay suggested that NFkappaB plays an important role in bortezomib suppressing FAK expression. SIGNIFICANCE: These results suggested that FAK is downregulated by bortezomib through a proteasome-dependent NFkappaB inhibitory mechanism. Thus, FAK could be a potential molecular target of bortezomib for therapeutic strategy. PMID- 20006626 TI - Overexpressed miR-494 down-regulates PTEN gene expression in cells transformed by anti-benzo(a)pyrene-trans-7,8-dihydrodiol-9,10-epoxide. AB - AIMS: We investigated the functionality of miR-494 in anti-benzo(a)pyrene-trans 7,8-dihydrodiol-9,10-epoxide (anti-BPDE)-transformed human bronchial epithelial cell 16HBE to reveal its potential target coding-gene. MAIN METHODS: The expression of mature miR-494 in cells was detected by miRNA-specific quantitative real-time polymerase chain reaction (QRT-PCR). QRT-PCR and Western blot were used to identify the expression of phosphatase and tensin homolog (PTEN) mRNA and protein. Following activation or inhibition of mature miRNA expression with precursors or antisense inhibitors, PTEN expression, luciferase activities, cell apoptosis, cell growth in soft agar and cell motility were analyzed. KEY FINDINGS: The expression of miR-494 increased while PTEN protein appeared to be lower in malignant transformed 16HBE cells. Enforced miR-494 level decreased PTEN protein expression compared to a negative precursor control group. Inhibition of miR-494 expression increased PTEN protein expression compared to negative inhibitor control group. Decreased expression of miR-494 increased caspase-3/7 activities in transformed 16HBE cells, and increased expression of miR-494 decreased this activity. Inhibition of miR-494 also decreased the malignancy of transformed cells. SIGNIFICANCE: MiR-494 regulates the expression of PTEN post transcriptionally and functions as a micro-oncogene in carcinogenesis induced by anti-BPDE. MiR-494 may be a useful target for gene therapy. PMID- 20006627 TI - Minocycline inhibits 5-lipoxygenase expression and accelerates functional recovery in chronic phase of focal cerebral ischemia in rats. AB - AIMS: We previously reported that minocycline attenuates acute brain injury and inflammation after focal cerebral ischemia, and this is partly mediated by inhibition of 5-lipoxygenase (5-LOX) expression. Here, we determined the protective effect of minocycline on chronic ischemic brain injury and its relation with the inhibition of 5-LOX expression after focal cerebral ischemia. MAIN METHODS: Focal cerebral ischemia was induced by 90 min of middle cerebral artery occlusion followed by reperfusion for 36 days. Minocycline (45 mg/kg) was administered intraperitoneally 2h and 12h after ischemia and then every 12h for 5 days. Sensorimotor function was evaluated 1-28 days after ischemia and cognitive function was determined 30-35 days after ischemia. Thereafter, infarct volume, neuron density, astrogliosis, and 5-LOX expression in the brain were determined. KEY FINDINGS: Minocycline accelerated the recovery of sensorimotor and cognitive functions, attenuated the loss of neuron density, and inhibited astrogliosis in the boundary zone around the ischemic core, but did not affect infarct volume. Minocycline significantly inhibited the increased 5-LOX expression in the proliferated astrocytes in the boundary zone, and in the macrophages/microglia in the ischemic core. SIGNIFICANCE: Minocycline accelerates functional recovery in the chronic phase of focal cerebral ischemia, which may be partly associated with the reduction of 5-LOX expression. PMID- 20006628 TI - Progressive associative phonagnosia: a neuropsychological analysis. AB - There are few detailed studies of impaired voice recognition, or phonagnosia. Here we describe two patients with progressive phonagnosia in the context of frontotemporal lobar degeneration. Patient QR presented with behavioural decline and increasing difficulty recognising familiar voices, while patient KL presented with progressive prosopagnosia. In a series of neuropsychological experiments we assessed the ability of QR and KL to recognise and judge the familiarity of voices, faces and proper names, to recognise vocal emotions, to perceive and discriminate voices, and to recognise environmental sounds and musical instruments. The patients were assessed in relation to a group of healthy age matched control subjects. QR exhibited severe impairments of voice identification and familiarity judgments with relatively preserved recognition of difficulty matched faces and environmental sounds; recognition of musical instruments was impaired, though better than recognition of voices. In contrast, patient KL exhibited severe impairments of both voice and face recognition, with relatively preserved recognition of musical instruments and environmental sounds. Both patients demonstrated preserved ability to analyse perceptual properties of voices and to recognise vocal emotions. The voice processing deficit in both patients could be characterised as associative phonagnosia: in the case of QR, this was relatively selective for voices, while in the case of KL, there was evidence for a multimodal impairment of person knowledge. The findings have implications for current cognitive models of voice recognition. PMID- 20006629 TI - Medication impairs probabilistic classification learning in Parkinson's disease. AB - In Parkinson's disease (PD), it is possible that tonic increase of dopamine associated with levodopa medication overshadows phasic release of dopamine, which is essential for learning. Thus while the motor symptoms of PD are improved with levodopa medication, learning would be disrupted. To test this hypothesis, we investigated the effect of levodopa medication on learning on the weather prediction task (WPT), which involves probabilistic classification learning. 11 PD patients and 13 matched controls completed 200 trials of the WPT, with the patients either on or off their usual levodopa medication. Consistent with prior studies, when PD patients were assessed on medication, overall WPT performance was significantly worse than controls. However, when these patients were studied following withdrawal from medication, overall performance was equivalent to controls, and significantly better than when on medication. The significant deterioration of learning on the WPT in PD patients when on compared to off medication supports the proposal that tonic increase of dopamine with dopaminergic medication masks phasic changes in dopamine release essential for learning. These results highlight the need for careful 'titration' of dopaminergic medication to produce the desired improvement of the motor symptoms without the associated detrimental effects on cognition and learning. PMID- 20006630 TI - Learning and forgetting new names and objects in MCI and AD. AB - We studied how subjects with mild cognitive impairment (MCI), early Alzheimer's disease (AD) and age-matched controls learned and maintained the names of unfamiliar objects that were trained with or without semantic support (object definitions). Naming performance, phonological cueing, incidental learning of the definitions and recognition of the objects were tested during follow-up. We found that word learning was significantly impaired in MCI and AD patients, whereas forgetting patterns were similar across groups. Semantic support showed a beneficial effect on object name retrieval in the MCI group 8 weeks after training, suggesting that the MCI patients' preserved semantic memory can compensate for impaired episodic memory. The MCI group performed equally well as the controls in the tasks measuring incidental learning and recognition memory, whereas the AD group showed impairment in this respect. Both the MCI and the AD group benefited less from phonological cueing than the controls. Our findings indicate that word learning is compromised in both MCI and AD, whereas long-term retention of newly learned words is not affected to the same extent. Incidental learning and recognition memory seem to be well preserved in MCI. PMID- 20006632 TI - Human thermoregulatory behavior during rest and exercise - a prospective review. AB - Despite the important role of temperature regulation in human behavior, it is frequently overlooked as a thermoregulatory response during both rest and exercise. During rest, the initiation of thermoregulatory behavior is preceded by changes in thermal comfort/sensation, with the temperature of the skin playing a vital signaling role. This behavior maintains heat balance and prevents the activation of autonomic thermoregulatory responses. Recently, self-paced exercise in the heat has been used as a thermo-behavioral model and accordingly, reductions in exercise work-rate in the heat appear sufficient to maintain regulation. Similar to rest, this behavior is mediated by elevations in skin temperature, however the perception of effort appears to be the perceptual trigger. PMID- 20006631 TI - Is the hippocampus necessary for visual and verbal binding in working memory? AB - A series of experiments test the recent claim that the hippocampus is necessary for the binding of features in working memory. Some potential limitations of studies underlying this claim are discussed, and an attempt is made to further test the hypothesis by studying a case of developmental amnesia whose extensively investigated pathology appears to be principally limited to the hippocampus, and who shows the expected deficit in episodic long-term memory. One series of experiments studied the short-term visual binding of color and shape under conditions ranging from simple presentation of colored objects through the more demanding task of combining the features when separated in space, or in time. A second set of experiments studied the capacity to use sentence structure to bind words into chunks in short-term verbal memory. Hippocampal pathology did not lead to a decrement on any of these tasks, suggesting that the hippocampus is not essential for short-term binding in working memory. PMID- 20006633 TI - Responsiveness to methamphetamine in adulthood is altered by prenatal exposure in rats. AB - Methamphetamine (MA) is a drug causing potent psychomotor activation. The aim of the present study was: (1) to assess the effect of prenatal and acute MA administration on behavior in adult male rats and (2) to find out if the prenatal exposure to MA increases sensitivity to acute MA application in adulthood. Behavior of adult male rats prenatally exposed to MA (5mg/kg) or saline was tested in Open field (OF) and Elevated plus maze (EPM). Subcutaneously administered MA (1mg/kg) or saline were used as challenge in adulthood, 30 min prior to testing. Our results showed that prenatal MA did not have an effect on baseline behavior in either of the tests. By contrast, acute MA increased overall psychomotor activity by increasing locomotion and exploratory behavior and decreasing comforting behavior. Moreover, adult rats prenatally exposed to MA exhibited increased sniffing and decreased rearing after acute MA dose in adulthood relative to prenatally saline-exposed rats. In addition, while acute MA application decreased anxiety in rats prenatally exposed to MA, rats prenatally exposed to saline were less sensitive to the anxiolytic effects of MA. Our results indicate that changes caused by prenatal exposure to psychostimulants may become apparent as different reactivity to drugs of abuse when an individual encounters them later in life. In addition, we found that the anxiolytic effect of acute MA (1mg/kg) probably depends also on the reactivity to stress and the activity of hypothalamo-pituitary-adrenal axis. PMID- 20006634 TI - Human health safety evaluation of cosmetics in the EU: a legally imposed challenge to science. AB - As stated in the European legislation, cosmetic products present on the European market must be safe for the consumer. Safety evaluation of the products is carried out by a qualified safety assessor who needs to consider potential exposure scenarios next to the physicochemical and toxicological profiles of all composing ingredients. Whereas, until recently, the tools to determine the toxicological profile of cosmetic ingredients mainly consisted of animal experiments, they have now been narrowed down substantially by the legally imposed animal testing ban on cosmetic ingredients, taken up in the Cosmetic Products Directive (76/768/EEC). This Directive, however, is not a stand-alone piece of European legislation, since as well directly as indirectly it is influenced by a complex web of related legislations. Vertical legislations deal with different categories of chemicals, including dangerous substances, biocides, plant protection products, food additives, medicinal products, and of course also cosmetics. Horizontal legislative texts, on the contrary, cover more general fields such as protection of experimental animals, consumer product safety, misleading of consumers, specific provisions for aerosols, and others. Experience has learnt that having a general overview of these related legislations is necessary to understand their impact on the cosmetic world in general terms and on cosmetic safety evaluation in particular. This goes for a variety of concerned parties, including national and European regulators/agencies, contract laboratories, raw material suppliers, cosmetic companies, research and educational centers. They all deal with a number of aspects important for the quality and toxicity of cosmetics and their ingredients. This review summarises the most relevant points of the legislative texts of different types of product categories and emphasises their impact on the safety evaluation of cosmetics. PMID- 20006635 TI - Arsenite suppression of involucrin transcription through AP1 promoter sites in cultured human keratinocytes. AB - While preserving keratinocyte proliferative ability, arsenite suppresses cellular differentiation markers by preventing utilization of AP1 transcriptional response elements. In present experiments, arsenite had a dramatic effect in electrophoretic mobility supershift analysis of proteins binding to an involucrin promoter AP1 response element. Without arsenite treatment, binding of JunB and Fra1 was readily detected in nuclear extracts from preconfluent cultures and was not detected a week after confluence, while c-Fos was detected only after confluence. By contrast, band shift of nuclear extracts from arsenite treated cultures showed only JunB and Fra1 binding in postconfluent as well as preconfluent cultures. Immunoblotting of cell extracts showed that arsenite treatment prevented the loss of Fra1 and the increase in c-Fos proteins that occurred after confluence in untreated cultures. Chromatin immunoprecipitation assays demonstrated substantial reduction of c-Fos and acetylated histone H3 at the proximal and distal AP1 response elements in the involucrin promoter and of coactivator p300 at the proximal element. Alteration of AP1 transcription factors was also examined in response to treatment with four metal containing compounds (chromate, vanadate, hemin, divalent cadmium) that also suppress involucrin transcription. These agents all influenced transcription at AP1 elements in a transcriptional reporter assay, but exhibited less effect than arsenite on binding activity assessed by mobility shift and chromatin immunoprecipitation and displayed variable effects on AP1 protein levels. These findings help trace a mechanism by which transcriptional effects of arsenite become manifest and help rationalize the unique action of arsenite, compared to the other agents, to preserve proliferative ability. PMID- 20006636 TI - Inorganic mercury causes pancreatic beta-cell death via the oxidative stress induced apoptotic and necrotic pathways. AB - Mercury is a well-known highly toxic metal. In this study, we characterize and investigate the cytotoxicity and its possible mechanisms of inorganic mercury in pancreatic beta-cells. Mercury chloride (HgCl2) dose-dependently decreased the function of insulin secretion and cell viability in pancreatic beta-cell-derived HIT-T15 cells and isolated mouse pancreatic islets. HgCl2 significantly increased ROS formation in HIT-T15 cells. Antioxidant N-acetylcysteine effectively reversed HgCl2-induced insulin secretion dysfunction in HIT-T15 cells and isolated mouse pancreatic islets. Moreover, HgCl2 increased sub-G1 hypodiploids and annexin-V binding in HIT-T15 cells, indicating that HgCl2 possessed ability in apoptosis induction. HgCl2 also displayed several features of mitochondria-dependent apoptotic signals including disruption of the mitochondrial membrane potential, increase of mitochondrial cytochrome c release and activations of poly (ADP ribose) polymerase (PARP) and caspase 3. Exposure of HIT-T15 cells to HgCl2 could significantly increase both apoptotic and necrotic cell populations by acridine orange/ethidium bromide dual staining. Meanwhile, HgCl2 could also trigger the depletion of intracellular ATP levels and increase the LDH release from HIT-T15 cells. These HgCl2-induced cell death-related signals could be significantly reversed by N-acetylcysteine. The intracellular mercury levels were markedly elevated in HgCl2-treated HIT-T15 cells. Taken together, these results suggest that HgCl2-induced oxidative stress causes pancreatic beta-cell dysfunction and cytotoxicity involved the co-existence of apoptotic and necrotic cell death. PMID- 20006639 TI - Focused issue on: novel approaches to protein crystallisation and crystal quality enhancement. PMID- 20006640 TI - Lifestyle behaviors and receipt of preventive health care services among hypertensive Americans aged 45 years or older in 2007. AB - OBJECTIVE.: The aim of the study was to explore opportunities for more effective interventions on lifestyle modifications among hypertensives. METHODS.: Using 2007 data from the Behavioral Risk Factor Surveillance System from the United States, we assessed the prevalence of various lifestyle behaviors and receipt of preventive health care services among US adults aged 45 years or older based on hypertension and treatment status (n=218,228). RESULTS.: Compared with nonhypertensives, hypertensives were significantly less likely to engage in recommended levels of physical activity and to be more obese regardless of treatment status. Compared to nonhypertensives, hypertensives on medical treatment were less likely to consume five or more servings of fruit and vegetables per day, be current smokers, and binge drinkers. Hypertensives not on medical treatment were more likely to be current smokers, binge and heavy drinkers. Hypertensives on medical treatment were more likely to have routine health checkup, have blood cholesterol checkup within previous 5 years and have a flu shot within the previous year than hypertensives not on treatment. CONCLUSION.: Different patterns of lifestyle behaviors and use of preventive health care services were found based on treatment status among hypertensives. Intervention programs should be tailored accordingly to achieve optimal outcome on lifestyle modification. PMID- 20006641 TI - Exercise adherence and intervention effects of two school-based resistance training programs for adolescents. AB - OBJECTIVE: The aim of this study was to evaluate the efficacy and feasibility of two school-based RT programs to improve muscular fitness and body composition in adolescents. METHODS: The study was conducted in Australia from July 2008 to June 2009. Participants [n=108, mean age (SD)=15.0 (0.7) years] were randomized to free weights (n=37) or elastic tubing (n=41) RT groups and a control group was recruited (n=30). Participants in the RT groups completed 2 sets of 10-12 repetitions on 10 exercises for 8 weeks. Waist circumference, body composition (bioelectrical impedance analysis) and muscular strength (bench press and leg press) were assessed at baseline and posttest. RESULTS: Boys (p<0.001) and girls (p<0.01) in both RT groups improved their body composition over the study period. Boys in both RT groups significantly improved both upper and lower body strength and their improvements were significantly greater than changes observed in the control group. Girls in the free weights groups achieved larger improvements in lower body strength compared to the control group (p<0.05). CONCLUSIONS: Free weights and elastic tubing RT are feasible and effective strategies for improving aspects of health-related fitness in adolescents. PMID- 20006638 TI - Roles of inositol phosphates and inositol pyrophosphates in development, cell signaling and nuclear processes. PMID- 20006642 TI - Telephone intervention promoting weight-related health behaviors. AB - OBJECTIVE: Recent national surveys have documented that the majority of adults in the United States do not meet the recommended levels of healthy lifestyle-related behaviors. The Nutrition and Physical Activity (NuPA) study was designed to promote fruit and vegetable consumption, physical activity, and weight management for a working population. METHOD: Data were collected nationwide, USA, from 2005 to 2007 and analyzed in 2008. A total of 2470 employed participants were randomized into the self-help (SH: n=1191) or self-help plus telephone counseling (SH+C: n=1279) group. The SH+C group received nine structured telephone counseling sessions in addition to the print materials. RESULTS: A series of hierarchical regression analyses for each of the health behavior outcomes in the present-at-follow-up subsample (n=1098-1148) revealed that the SH+C was effective in increasing fruit and vegetable consumption. Among the overweight and obese participants, weight loss was significant in both the SH and SH+C groups. CONCLUSION: Using a theory-based behavioral change counseling technique and targeting multiple health behaviors among employed individuals, our findings demonstrate that the addition of telephone counseling to mailed self-help materials is effective in promoting healthy diet and weight management. PMID- 20006643 TI - Economic considerations of HPV vaccination. PMID- 20006645 TI - Control mechanisms in working memory: a possible function of EEG theta oscillations. AB - Neural correlates of control mechanisms in human working memory are discussed at two levels in this review: (i) at 'item level', where in multi-item working memory information needs to be organized into sequential memory representations, and (ii) at a 'process level', indicating the integration and control of a variety of cognitive functions involved in working memory, independent of item representations per se. It will be discussed that at both levels electroencephalographic theta activity is responsible for control of working memory functions. On item level, exact phase coding, e.g., approached by coupling between theta and gamma oscillations or phase resetting of theta frequency, is suggested to integrate information into working memory representations. At process level interregional theta synchronization is discussed to integrate brain structures necessary for working memory. When discussing the specificity of theta activity for control of working memory processes it will be suggested that theta oscillations might play an important general integrative role in organization of brain activity. And as working memory often involves a variety of cognitive processes which need to be coordinated there is particular need for an integrative brain mechanism like theta activity as suggested in this review. PMID- 20006644 TI - Colorectal cancer screening barriers and facilitators in older persons. AB - BACKGROUND: This systematic review identifies factors that are most consistently mentioned as either barriers to or facilitators of colorectal cancer (CRC) screening in older persons. METHODS: A systematic literature search (1995-2008) was conducted to identify studies that reported barriers to or facilitators of CRC screening uptake, compliance or adherence specifically for older persons (> or = 65 years). Information on study characteristics and barriers and facilitators related to subjects; healthcare providers; policies; and screening tests were then abstracted and analyzed. RESULTS: Eighty-three studies met the eligibility criteria. Low level of education, African American race, Hispanic ethnicity, and female gender were the most frequently reported barriers, whereas being married or living with a partner was the most frequently reported facilitator. The most cited barrier related to healthcare providers was lack of screening recommendation by a physician; having a usual source of care was a commonly reported facilitator. Lack of health insurance, and dual coverage with Medicare and Medicaid were the most frequently reported barriers, whereas Medicare's coverage of screening colonoscopy was consistently reported as a facilitator. CONCLUSIONS: Barriers to, and facilitators of, CRC screening among older persons are reported. Particular attention should be paid to modifiable factors that could become the focus of interventions aimed at increasing CRC screening participation in older persons. PMID- 20006646 TI - Implication of 5-HT2 receptor subtypes in the mechanism of action of the GABAergic compound etifoxine in the four-plate test in Swiss mice. AB - Etifoxine is an anxiolytic compound structurally unrelated to benzodiazepine and neurosteroids but potentiating GABA(A) receptor function by a dual mode of action including a direct positive allosteric modulation through a site distinct from that of benzodiazepines. Etifoxine has been shown to possess some anxiolytic-like effects in rodents. METHODS: Using the four-plate test (FPT) model of anxiety in mice the potential anxiolytic-like effect of etifoxine was first to re-evaluate. In a second part, in order to better understand the mechanism of action of etifoxine, interaction studies with 5-HT(2) ligands were performed in the FPT as mixed serotonergic and GABAergic mechanisms are highly implicated in the anxiolytic-like effect observed in the FPT. RESULTS: A dose response effect was observed for etifoxine from the dose of 40-100 mg/kg. Doses above to 60 mg/kg induced a sedative effect as was determined in the actimeter test. The 5-HT(2A) receptor antagonist SR 46349B blocked the anti-punishment activity of etifoxine (40 and 50 mg/kg), while the 5-HT(2B/2C) receptor antagonist, SB 206553 and the 5 HT(2C) receptor antagonist, RS 10-2221 did not alter its effects. In a same way, only the 5-HT(2A) agonist DOI induced anti-punishment effect when co-administered with subthreshold doses of etifoxine. CONCLUSION: The present results demonstrated that etifoxine effect was modulated by 5-HT(2A) ligands co administration. The large literature concerning GABA and 5-HT suggests that they could be co-released and could act as co-transmitters in some regions of the CNS and cross-communication between the two neurotransmitters might be an important modulator process of neuronal activity. PMID- 20006647 TI - Disrupted startle modulation in animal models for affective disorders. AB - Affective startle modulation is used to study emotional reactivity in humans, and blunted affective startle modulation has been reported in depressed patients. To determine whether blunted affective startle modulation is also a common feature in animal models for affective disorders, light-enhanced startle was studied in three models: inescapable foot shock (IFS), repeated restraint stress (RRS) and olfactory bulbectomy (OBX). In addition, prepulse inhibition was studied in these models. Light-enhanced startle was blunted following IFS and OBX and RRS decreased overall startle responding. Prepulse inhibition, however, was unaffected. These findings indicate that induction models for affective disorders may be associated with long term effects on affective startle modulation. The lack of changes in sensory motor gating suggests that these changes can be ascribed to alterations in emotional reactivity. In conclusion, our results indicate that the blunted affective startle modulation seen in animal models for affective disorders may be used to examine the mechanisms underlying altered emotional reactivity. PMID- 20006648 TI - The modified Geller-Seifter test in rats was insensitive to GABAB receptor positive modulation or blockade, or 5-HT1A receptor activation. AB - RATIONALE: Both the GABA(B) receptor positive modulator GS39783 and the GABA(B) receptor antagonist CGP46381 exhibit anxiolytic-like properties in animal models. In the present studies, the effects of GS39783 and CGP46381 in the modified Geller-Seifter task were assessed. First, the predictive validity of the task was confirmed by assessing the effects of multiple anxiolytic and non-anxiolytic compounds on punished and unpunished responding. METHODS: Rats were trained in the modified Geller-Seifter task. After successful acquisition of the task, chlordiazepoxide, diazepam, MPEP, haloperidol, GS39783, 8-OH-DPAT, alprazolam and CGP46381 were tested consecutively. For each test compound, doses were administered in a randomized, counter-balanced, within-subjects design. Drug tests were performed only when rats exhibited baseline performance (the punished and time-out response rates were less than 10% of the unpunished response rate). RESULTS: Chlordiazepoxide, diazepam, alprazolam and MPEP released punished responding with variable effects on unpunished responding. Haloperidol had a small but significant effect on punished responding at an intermediate dose, and decreased unpunished responding at the highest dose tested. In contrast, administration of the GABA(B) receptor positive modulator GS398783 or the GABA(B) receptor antagonist CGP46381 at doses up to 30 mg/kg had no effects on either punished or unpunished responding. The 5-HT(1A) agonist 8-OH-DPAT did not release punished responding, but significantly decreased unpunished responding at the highest dose tested. SUMMARY: The modified Geller-Seifter task generally exhibits good predictive validity for anxiolytic-like compounds. Neither GABA(B) receptor positive allosteric modulation nor blockade exhibited anxiolytic-like properties in the modified Geller-Seifter task. The 5-HT(1A) partial agonist buspirone was similarly ineffective. PMID- 20006649 TI - Influence of sound specificity and familiarity on Japanese macaques' (Macaca fuscata) auditory laterality. AB - Despite attempts to generalise the left hemisphere-speech association of humans to animal communication, the debate remains open. More studies on primates are needed to explore the potential effects of sound specificity and familiarity. Familiar and non-familiar nonhuman primate contact calls, bird calls and non biological sounds were broadcast to Japanese macaques. Macaques turned their heads preferentially towards the left (right hemisphere) when hearing conspecific or familiar primates supporting hemispheric specialisation. Our results support the role of experience in brain organisation and the importance of social factors to understand laterality evolution. PMID- 20006651 TI - Modeling withdrawal syndrome in zebrafish. AB - The zebrafish (Danio rerio) is rapidly becoming a popular model species in behavioral neuroscience research. Zebrafish behavior is robustly affected by environmental and pharmacological manipulations, and can be examined using exploration-based paradigms, paralleled by analysis of endocrine (cortisol) stress responses. Discontinuation of various psychotropic drugs evokes withdrawal in both humans and rodents, characterized by increased anxiety. Sensitivity of zebrafish to drugs of abuse has been recently reported in the literature. Here we examine the effects of ethanol, diazepam, morphine and caffeine withdrawal on zebrafish behavior. Overall, discontinuation of ethanol, diazepam and morphine produced anxiogenic-like behavioral or endocrine responses, demonstrating the utility of zebrafish in translational research of withdrawal syndrome. PMID- 20006652 TI - var gene transcription dynamics in Plasmodium falciparum patient isolates. AB - A major feature of Plasmodium falciparum parasitized red blood cells (pRBC) is their capacity to sequester in the microcirculation. The binding is mediated by PfEMP1 (P. falciparum erythrocyte membrane protein 1), a variable protein encoded by the var gene family. P. falciparum avoids the host antibody response generated against previously used variants by switching the expression of PfEMP1, which may affect the disease outcome. We have here studied var gene transcription over time within the life cycle of the parasite by semi-quantitative PCR and sequencing by employing three sets of degenerate primers to the 5-prime end of the var genes (corresponding to the DBL1alpha-domain). To accurately determine transcript levels, subsequent in-depth analysis was made by amplifying the 10 most frequently expressed var sequences identified in each developmental stage by quantitative PCR (Q-PCR). The maximum peak in var gene transcription seems to vary in time among parasites. In five out of seven parasites, var gene transcription was found to be higher or equal at 22-26h post-invasion compared to 4-10h post-invasion. Our data indicate that the intra-isolate var gene transcription dominance order may change between different developmental stages. The transcription of var genes in field isolates is more complex than in laboratory strains and often changes after in vitro adaption of the parasite. By using semi-quantitative PCR employing degenerate primers combined with quantitative-PCR using specific primers it is possible to monitor var gene transcription in detail during the life cycle of the parasite. The work presented here suggests that trophozoite pRBC is likely to be the optimal source of RNA for predicting the translated var gene species. PMID- 20006650 TI - Behavioral analysis of Ste20 kinase SPAK knockout mice. AB - SPAK/STK39 is a mammalian protein kinase involved in the regulation of inorganic ion transport mechanisms known to modulate GABAergic neurotransmission in the both central and the peripheral nervous systems. We have previously shown that disruption of the gene encoding SPAK by homologous recombination in mouse embryonic stem cells results in viable mice that lack expression of the kinase. With the exception of reduced fertility, these mice do not exhibit an overt adverse phenotype. In the present study, we examine the neurological phenotype of these mice by subjecting them to an array of behavioral tests. We show that SPAK knockout mice displayed a higher nociceptive threshold than their wild-type counterparts on the hot plate and tail flick assays. SPAK knockout mice also exhibited a strong locomotor phenotype evidenced by significant deficits on the rotarod and decreased activity in open-field tests. In contrast, balance and proprioception was not affected. Finally, they demonstrated an increased anxiety like phenotype, spending significantly longer periods of time in the dark area of the light/dark box and increased thigmotaxis in the open-field chamber. These results suggest that the kinase plays an important role in CNS function, consistent with SPAK regulating ion transport mechanisms directly involved in inhibitory neurotransmission. PMID- 20006653 TI - Chromogranin A: a new proposal for trafficking, processing and induction of granule biogenesis. AB - Chromogranin A (CgA), a member of the granin family serves several important cell biological roles in (neuro)endocrine cells which are summarized in this review. CgA is a "prohormone" that is synthesized at the rough endoplasmic reticulum and transported into the cisternae of this organelle via its signal peptide. It is then trafficked to the Golgi complex and then to the trans-Golgi network (TGN) where CgA aggregates at low pH in the presence of calcium. The CgA aggregates provide the physical driving force to induce budding of the TGN membrane resulting in dense core granule (DCG) formation. Within the granule, a small amount of the CgA is processed to bioactive peptides, including a predicted C terminal peptide, serpinin. Upon stimulation, DCGs undergo exocytosis and CgA and its derived peptides are released. Serpinin, acting extracellularly is able to signal the increase in transcription of a serine protease inhibitor, protease nexin-1 (PN-1) that protects DCG proteins against degradation in the Golgi complex, which then enhances DCG biogenesis to replenish those that were released. Thus CgA and its derived peptide, serpinin, plays a significant role in granule formation and regulation of granule biogenesis, respectively, in (neuro) endocrine cells. PMID- 20006654 TI - Secretoneurin as a hormone regulator in the pituitary. AB - Secretoneurin (SN) is a 33-34 amino acid peptide derived from the most conserved sequence of the secretogranin (SgII) precursor. SgII is a granin protein found in the secretory granules of neuroendocrine tissues. There are two paralogs of teleost SgII that we name here SgIIa and SgIIb. Processing of these proteins would yield SNa and SNb in fish. Secretoneurin immunoreactivity is found within all the major pituitary cell types in mammals. In goldfish, it appears to be mainly expressed in the prolactin cells of the rostral pars distalis. We have investigated the paracrine role of goldfish SN (SNa) to stimulate luteinizing hormone from gonadotrophs in the neighboring proximal pars distalis. Another source of SN is the hypophysiotropic neurons that may deliver SN to target cells by direct pituitary innervation. Little else is known about the neuroendocrine role of SN. We also discuss the evolution, distribution and production of SN in the pituitary. PMID- 20006655 TI - Molecular chaperones, essential partners of steroid hormone receptors for activity and mobility. AB - Steroid hormone receptors (SHRs) are notorious intracellular travellers, transiting among different cellular compartments as they mature, are subjected to regulation and exert their biological functions. Understanding the processes governing the intracellular traffic of SHRs is important, since their unbalanced or erroneous localization could lead to the development of diseases. In this review, we not only explore the functions of the heat-shock protein 90 (Hsp90) molecular chaperone machine for the intracellular transport of SHRs, but also for the regulation of their nuclear mobility, for their recycling and for the regulation of their transcriptional output. PMID- 20006656 TI - A rapid multiplex assay for nucleic acid-based diagnostics. AB - We have developed a rapid (under 4 hours), multiplex, nucleic acid assay, adapted to a microsphere array detection platform. We call this assay multiplex oligonucleotide ligation-PCR (MOL-PCR). Unlike other ligation-based assays that require multiple steps, our protocol consists of a single tube reaction, followed by hybridization to a Luminex microsphere array for detection. We demonstrate the ability of this assay to simultaneously detect diverse nucleic acid signatures (e.g., unique sequences, single nucleotide polymorphisms) in a single multiplex reaction. Detection probes consist of modular components that enable target detection, probe amplification, and subsequent capture onto microsphere arrays. To demonstrate the utility of our assay, we applied it to the detection of three biothreat agents, B. anthracis, Y. pestis, and F. tularensis. Combined with the ease and robustness of this assay, the results presented here show a strong potential of our assay for use in diagnostics and surveillance. PMID- 20006657 TI - Hyperactivity in novel environment with increased dopamine and impaired novelty preference in apoptosis signal-regulating kinase 1 (ASK1)-deficient mice. AB - Apoptosis signal-regulating kinase 1 (ASK1) is a mitogen-activated protein (MAP) kinase kinase kinase family member, which induces apoptosis in various cells through JNK and p38 MAP kinase cascades. In addition to apoptosis signaling, a number of recent in vitro studies have suggested that ASK1 may play roles in neural function. However, the behavioral significance of ASK1 has remained unclear. Here, we subjected ASK1 (-/-) mice to a battery of behavioral tests and found that they displayed temporary hyperactivity in an open-field test. Activities in the familiar field were normal, indicating that the hyperactivity observed was specific to the novel environment. ASK1 (-/-) mice also exhibited impairment of novelty preference 24h after training and superior performance on the rotarod test. Brain tissue contents of dopamine and 4-dihydroxyphenylacetic acid (DOPAC) were elevated in ASK1 (-/-) mice. Our findings thus demonstrate novel behavioral functions of ASK1, including regulation of locomotor activity, novelty preference, and motor coordination with dopaminergic transmission. PMID- 20006658 TI - Development of the somatosensory cortex, the cerebellum, and the main olfactory system in Semaphorin 3F knockout mice. AB - Semaphorin 3F (Sema3F) is a secreted type of the semaphorin family of axon guidance molecules. Sema3F and its receptor Neuropilin-2 (Npn-2) mRNAs were distributed in a mutually exclusive manner throughout mouse brain development. In order to examine physiological roles of Sema3F, we generated Sema3F knockout mice (KO) by gene targeting in embryonic stem (ES) cells. We found that the loss of Sema3F expression did not significantly affect the mRNA expression of Npn-2 or the other putative Npn-2 ligands, namely, Sema3B, Sema3C, or Sema3G. The barrel structure of the somatosensory cortex and the cerebellar neuroanatomy were not significantly altered in Sema3F KO. Finally, optical imaging of intrinsic signals of the dorsal olfactory bulb showed no significant differences in odor map between wild-type mice and Sema3F KO. These data suggest that Sema3F plays a relatively restricted, if any, role in its receptor expression and postnatal development of these brain structures. PMID- 20006659 TI - An optical and microPET assessment of thermally-sensitive liposome biodistribution in the Met-1 tumor model: Importance of formulation. AB - The design of delivery vehicles that are stable in circulation but can be activated by exogenous energy sources is challenging. Our goals are to validate new imaging methods for the assessment of particle stability, to engineer stable and activatable particles and to assess accumulation of a hydrophilic model drug in an orthotopic tumor. Here, liposomes were injected into the tail vein of FVB mice containing bilateral Met-1 tumors and imaged in vivo using microPET and optical imaging techniques. Cryo-electron microscopy was applied to assess particle shape prior to injection, ex vivo fluorescence images of dissected tissues were acquired, excised tissue was further processed with a cell-digest preparation and assayed for fluorescence. We find that for a stable particle, in vivo tumor images of a hydrophilic model drug were highly correlated with PET images of the particle shell and ex vivo fluorescence images of processed tissue, R(2)=0.95 and R(2)=0.99 respectively. We demonstrate that the accumulation of a hydrophilic model drug is increased by up to 177 fold by liposomal encapsulation, as compared to accumulation of the drug at 24 hours. PMID- 20006660 TI - Regulated non-viral gene delivery from coaxial electrospun fiber mesh scaffolds. AB - In an effort to add to the versatility of three-dimensional scaffolds for tissue engineering applications, recent experimental designs are incorporating biological molecules such as plasmids and proteins within the scaffold structure. Such scaffolds act as reservoirs for the biological molecules of interest while regulating their release over various durations of time. Here, we describe the use of coaxial electrospinning as a means for the fabrication of fiber mesh scaffolds and the encapsulation and subsequent release of a non-viral gene delivery vector over a period of up to 60 days. Various fiber mesh scaffolds containing plasmid DNA (pDNA) within the core and the non-viral gene delivery vector poly(ethylenimine)-hyaluronic acid (PEI-HA) within the sheath of coaxial fibers were fabricated based on a fractional factorial design that investigated the effects of four processing parameters at two levels. Poly(epsilon caprolactone) sheath polymer concentration, poly(ethylene glycol) core polymer molecular weight and concentration, and the concentration of pDNA were investigated for their effects on average fiber diameter, release kinetics of PEI HA, and transfection efficiency. It was determined that increasing the values of each of the investigated parameters caused an increase in the average diameter of the fibers. The release kinetics of PEI-HA from the fibers were affected by the loading concentration of pDNA (with PEI-HA concentration adjusted accordingly to maintain a constant nitrogen to phosphorous (N:P) ratio within the complexes). Two-dimensional cell culture experiments with model fibroblast-like cells demonstrated that complexes of pDNA with PEI-HA released from fiber mesh scaffolds could successfully transfect cells and induce expression of enhanced green fluorescent protein (EGFP). Peak EGFP expression varied with the investigated processing parameters, and the average transfection observed was a function of poly(ethylene glycol) (core) molecular weight and concentration. Furthermore, fibroblast-like cells seeded directly onto coaxial fiber mesh scaffolds containing PEI-HA and pDNA showed EGFP expression over 60 days, which was significantly greater than the EGFP expression observed with scaffolds containing pDNA alone. Hence, variable transfection activity can be achieved over extended periods of time upon release of pDNA and non-viral gene delivery vectors from electrospun coaxial fiber mesh scaffolds, with release and subsequent transfection controlled by tunable coaxial fiber mesh fabrication parameters. PMID- 20006661 TI - Olfactory priming reinstates extinguished chocolate-induced conditioned place preference. AB - A major problem in the dietary treatment of disorders associated with excessive eating, such as obesity, is the high rate of relapse into maladaptive eating habits after withdrawal from consumption of palatable, energy-dense food. As olfaction has a major role in appetite and eating behavior, in this study we used a reinstatement model based on conditioned place preference to investigate the ability of olfactory priming to reinstate extinguished chocolate-induced conditioned place preference in sated mice. We found that olfactory priming, which was ineffective in inducing conditioned place preference in the control group, reactivated place preference following the extinction procedure in the experimental group. These results extend previous reports of the reinstatement of food seeking induced by pellet priming and, for the first time, show the possibility of using olfactory priming in an animal model of relapse. In light of the major role of olfactory inputs in appetite and of cues in relapse, the present results indicate that smell is an important factor to consider in the treatment of eating disorders. PMID- 20006662 TI - Are dietary restraint scales valid measures of dietary restriction? Additional objective behavioral and biological data suggest not. AB - Prospective studies find that individuals with elevated dietary restraint scores are at increased risk for bulimic symptom onset, yet experiments find that assignment to energy-deficit diet interventions reduce bulimic symptoms. One explanation for the conflicting findings is that the dietary restraint scales used in the former studies do not actually identify individuals who are restraining their caloric intake. Thus, we tested whether dietary restraint scales showed inverse relations to objectively measured caloric intake in three studies. Four dietary restraint scales did not correlate with doubly labeled water estimates of caloric intake over a 2-week period (M, r=.01). One scale showed a significant inverse correlation with objectively measured caloric intake during a regular meal ordered from an ecologically valid menu (M, r=-.30), but a significant positive relation that was qualified by a significant quadratic effect, to objectively measured caloric intake during multiple eating episodes in the lab (M, r=.32). In balance, results suggest that dietary restraint scales are not valid measures of dietary restriction, replicating findings from prior studies that examined objective measures of caloric intake. PMID- 20006664 TI - Expression of human beta-defensin-2 in intratumoral vascular endothelium and in endothelial cells induced by transforming growth factor beta. AB - Human beta-defensin-2 (hBD-2) is a small cationic peptide originally identified from psoriatic skin lesions as an antimicrobial agent of the innate immune system. The expression of hBD-2 is believed to be induced exclusively in epithelial cells by microbial components and certain proinflammatory cytokines, such as interleukin-1 beta (IL-1 beta). In this study, we report, for the first time, that hBD-2 is expressed in vascular endothelial cells associated with oral squamous cell carcinoma (OSCC) and Kaposi's sarcoma lesions, but not in that of normal stroma. Expression of hBD-2 in vascular endothelial cells was further substantiated by in vitro experiments using cultured human umbilical vein endothelial cells (HUVECs). Transforming growth factor beta1 (TGF beta 1) and IL 1 beta, two well-known tumorigenic inflammatory mediators, induce hBD-2 transcript and peptide expression in HUVECs. However, TGF beta 1 does not stimulate hBD-2 expression in oral epithelial cells. In addition, proinflammatory cytokines and microbial reagents do not induce the expression of hBD-1 and hBD-3 in HUVECs. Since hBD-2 has been shown to modulate migration, proliferation, and tube formation of HUVECs in vitro and participate in immune cell trafficking, its expression in vascular endothelial cells located within malignant lesions may play a role in tumor angiogenesis and cancer metastasis. PMID- 20006663 TI - An analysis of licking microstructure in three strains of mice. AB - Mouse models of feeding provide a useful tool for elucidating the molecular pathways of energy regulation. The majority of studies in mice have been limited to intake analyses conducted over extended periods of time, which fail to distinguish between a variety of factors that influence nutrient intake. Using licking microstructure analyses we examined both the size and number of licking bursts for water, polycose, sucrose and lecithin in three strains of mice (C57BL/6J, 129Sv/ImJ and C57129F1 hybrids), using pause criteria (250-500, >500 and >1000 ms) that have previously been described in the rat. Burst size and number varied both as a function of tastant concentration and mouse strain; however, these differences were most evident with the >1000 ms pause criterion. Consistent with previous reports, during water consumption C57 mice showed longer mean interlick intervals, a larger number of bursts but reduced burst size relative to the two other strains. F1 mice showed larger burst sizes for polycose, while C57 mice displayed a greater number of bursts for both polycose and sucrose. Both 129 and F1 mice were insensitive to sucrose concentration, whereas C57 mice showed attenuated lecithin intake influenced by a reduction in the size of bursts for this tastant. These results suggest that these strains of mice display differences in the pattern of licking that are most evident with the use of larger pause criteria. These differences in licking behavior might reflect influences of genetic background on pre- and post-ingestive factors controlling intake, the reinforcing properties of each tastant, or native differences in licking style. PMID- 20006665 TI - Biological activities and molecular interactions of the C-terminal residue of thrombospondin-4, an epitome of acidic amphipathic peptides. AB - C21, the C-terminal residue of thrombospondin-4 (TSP-4), was identified as a peptide growth factor during an investigation concerning erythropoietin dependent, erythroid stimulating factors of endothelial origin. It is active in cultures of several human hematopoietic stem cells, skin fibroblasts and kidney epithelial cells and stimulates red cell formation in anemic mice. A method of affinity chromatography in the presence of high concentrations of Triton X-100, previously developed for identifying proteins associated with the TSP-1 receptor CD47, was utilized for the detection of C21 binding molecules and their detergent resistant, associated partners. These experiments helped to delineate two different mechanisms of C21 action, which are compatible with its cell proliferating activity. As a cell matrix peptide, C21 binds to the osteopontin receptor CD44 and could act as an osteopontin antagonist, preventing the inhibition of primitive hematopoietic stem cell proliferation. TSP-1, another matrix protein, binds to C21 and could indirectly act as an antagonist, by shunting C21-CD44 interactions. The second mechanism is a direct effect of C21 on cell proliferation. The extremely rapid internalization and nuclear localization of the peptide could be explained by CD44-mediated internalization, followed by a microtubule-mediated transport towards the nucleus, or, eventually, direct membrane insertion. These alternative hypotheses are supported by previously observed membrane insertion of similar synthetic and viral acidic amphipathic peptides, the presence of microtubule-associated protein 1B (MAP1B) and dynactin in the triton-soluble complexes associated with C21 and the presence in such complexes of dual compartment proteins for nuclei and plasma membranes, such as MAP1B, AHNAK and CD44. PMID- 20006666 TI - Protein synthesis dependent effects of kinins on astrocyte prostaglandin synthesis. AB - It has been shown that kinins and their receptors are over expressed in the brain under pathophysiological conditions such as inflammation. However, little is known about the possible role of kinins, and especially bradykinin in brain inflammation. Although kinins are thought to have immediate effects, peptides may also exert longer and protein synthesis dependent actions. To evaluate this possibility, we assessed the regulation of prostaglandin E(2) synthesis after 15h bradykinin or Lys-des-Arg(9)-bradykinin (B(1) receptor agonist) treatment in rat neonatal astrocytes. Bradykinin, dose dependently stimulated basal and lipopolysaccharide-induced prostaglandin E(2) production, whereas exposure of astrocytes to the B(1) receptor agonist decreased both basal and lipopolysaccharide-induced prostaglandin E(2) release in a dose-dependent manner. These kinin effects on PGE(2) synthesis were completely abrogated by actinomycin D and cycloheximide, suggesting de novo synthesis of proteins. Bradykinin also increased cyclooxygenase-2 protein levels about 2-fold, while the B(1) receptor agonist decreased cyclooxygenase-2 protein expression. There was no change in cyclooxygenase-1 protein levels after treatment with either of the kinins. Our data suggest a delayed feedback regulatory mechanism of kinins on astrocyte inflammation, whereby astrocyte prostaglandin synthesis is initially enhanced by bradykinin (B(2)) and eventually blocked by kinin breakdown product, acting on B(1) receptors. At least part of this presumed feedback loop could be mediated by de novo protein synthesis of cyclooxygenase-2. PMID- 20006667 TI - Liver transplant recipients weaned off immunosuppression lack circulating donor specific antibodies. AB - Human leukocyte antigen (HLA)-specific antibodies (Abs) were examined in 73 clinically stable liver transplant recipients divided into group A (n = 19; clinically tolerant), group B (n = 34; undergoing weaning, on minimal immunosuppression), and group C (n = 20; had failed drug withdrawal or weaning never attempted). Of 19 patients in group A, six (32%) had anti-HLA Abs; none were donor-specific. In contrast, 23 of 34 patients (67%) in group B and nine of 20 patients (45%) in group C exhibited anti-HLA Abs (p = 0.02). Furthermore, 15 of 19 patients in groups B and C (9/12, p = 0.01 and 6/7, p = 0.01, respectively) exhibited donor-specific anti-HLA Abs. The prevalence of donor-specific HLA Abs was significantly higher in nontolerant patients. Five years after initial evaluation, >90% (18/19) group A patients remained off immunosuppression. One of seven of these patients available for retesting exhibited donor-specific Abs. In group B, two-fourths of 34 patients (12%) weaned successfully were HLA-Ab negative; four patients who experienced rejection while undergoing weaning exhibited anti-HLA Ab initially and at 5 years. Thus, most of the liver recipients off immunosuppression lacked donor-specific alloAbs. The occurrence of these alloAbs should now be examined prospectively in a drug weaning trial. PMID- 20006668 TI - Medical surveillance, exposure registries, and epidemiologic research for workers exposed to nanomaterials. AB - While there is a growing body of information about hazards of nanomaterials, little is known about the risks to workers exposed to them. However, workers are the first people in society that are being exposed to the growing inventory of "nano-enabled" products in commerce. The number of workers involved in the investigation, manufacture, production, and disposal of these types of products is growing. Although toxicologic research is still the highest priority, it is time to actively anticipate the health needs of workers. To date, precautionary risk management approaches have been widely advocated. Now there is a need to initiate an evolving process to identify the issues in medical surveillance, utilization of exposure registries, and the conduct of epidemiologic research. Each of these are related complex endeavors that build on the toxicologic evidence and extent of exposure. There is a need to assess the scientific basis and research needs for determining early functional changes, organ system and disease responses for use in targeted medical surveillance. There is also need for development of criteria for extrapolating toxicological data in biological systems to predict the risk of adverse outcomes in humans. In the meantime, exposure registries may be pivotal in helping societies act in the face of uncertainty in a precautionary manner, but legal, ethical, and logistical issues need resolution. Epidemiologic research will build on these efforts and may ultimately contribute critical definitive rationale for medical screening, risk assessment and management. PMID- 20006669 TI - The efficacy and safety of articaine versus lignocaine in dental treatments: a meta-analysis. AB - OBJECTIVES: Although articaine has been recommended for providing an improved local anaesthetic effect in patients presenting for dental treatments, a relevant meta-analysis has been lacking. Despite articaine's popularity, there is contradictory evidence to support the claims. The aim of this systematic review was to compare the efficacy and safety of articaine with lignocaine in maxillary and mandibular infiltrations and block anaesthesia in patients presenting for routine dental treatments. DATA SOURCES: The following databases were searched: Cochrane Central, Medline, Embase, and ProQuest Health and Medical Complete. In addition, the metaRegister of the controlled trials database was searched to identify dissertations and ongoing or unpublished trials, and the Australian division of Septodont (the manufacturer of articaine and lignocaine) was contacted. The bibliographies of identified articles were also searched. STUDY SELECTION: Inclusion was limited to: (1) randomized controlled trials in patients requiring non-complex routine dental treatments; (2) interventions comparing 4% articaine (1:100,000 epinephrine) with 2% lignocaine (1:100,000 epinephrine) for maxillary and mandibular infiltrations and block anaesthesia; and (3) with principal outcome measures of anaesthetic success, post-injection adverse events or post-injection pain. Trial quality was evaluated by assessing randomization, allocation concealment, blinding, intention to treat analyses and how losses to follow up were addressed. Treatment effects were combined by meta-analysis using the random effects method. RESULTS: Articaine is more likely than lignocaine to achieve an anaesthetic success in the posterior first molar area with a relative risk for success at 1.31 (95% CI 1.12-1.54, P=0.0009). There is no difference in post-injection adverse events between articaine and lignocaine with a relative risk of 1.05 (95% CI 0.66-1.65, P=0.85). However, articaine injection results in a higher pain score as measured by Visual Analogue Scale, than lignocaine at the injection site after anaesthetic reversal with a weighted mean difference of 6.49 (95% CI 0.02-12.96, P=0.05) decreasing to 1.10 (95% CI 0.18-2.02, P=0.02) on the third day after injection. CONCLUSION: The results of this systematic review provide support for the argument that articaine is more effective than lignocaine in providing anaesthetic success in the first molar region for routine dental procedures. In addition, both drugs appear to have similar adverse effect profiles. The clinical impact of articaine's higher post-injection pain scores than lignocaine is negligible. Hence, articaine is a superior anaesthetic to lignocaine for use in routine dental procedures. Use in children under 4 years of age is not recommended, since no data exists to support such usage. PMID- 20006670 TI - Conformational stability and energetics of Plasmodium falciparum glutaredoxin. AB - Glutaredoxins (Grxs), redox-active proteins with a typical -CPYC motif at their active sites, are involved in redox-regulatory processes and antioxidant defenses. The human malarial parasite Plasmodium falciparum possess a classical glutaredoxin (PfGrx) as well as a number of Grx-like proteins. In the present study, we investigated the unfolding energetics and conformational stability of PfGrx, using isothermal guanidine hydrochloride-induced and pH-dependent thermal denaturation. Reversible unfolding can be modeled using a two-state transition between the native and unfolded states. The structural topology of the protein was stable over a wide pH range from 3.0 to 11.0. Although the protein was thermally stable, it exhibited a small free energy of 1.56 kcal mol(-1) at 25 degrees C. The thermostability of PfGrx reached its maximum at pH 8.0, with a T(m) of 76.2 degrees C and a DeltaH(m) of 119 kcal mol(-1). To elucidate the factors underlying the thermostability, a protein stability curve was generated. Maximum stability occurred at around 47 degrees C, where the DeltaG_H2O(D) value was 4.30 kcal mol(-1). The high structural stability over a broad pH range, together with the capacity to endure very high temperatures, supports the notion that Grx can withstand a wide variety of conditions, allowing it to play a key role in cellular redox homeostasis. To the best of our knowledge, this work represents the first attempt to understand the energetic characteristics of a glutaredoxin in relation to accompanying structural changes. PMID- 20006672 TI - Endocrine aspects of obesity. PMID- 20006671 TI - Fast intra-axonal transport: Beginning, development and post-genome advances. AB - The review describes the initial experiments suggesting a fast intra-axonal transport of transmitter related substances, in addition to the "classic" slow flow. Early experiments were mainly conducted in the peripheral adrenergic system, focusing on transport of amine storage granules, the extent of the vast sympathetic adrenergic system and the importance of axonal transport of amine granules for the adrenergic system. Further, it describes important advances obtained from studies of other neuron systems regarding local axonal protein synthesis, motor proteins and new insights regarding relation between faults in the transport machinery and some neuropathological conditions. PMID- 20006673 TI - Helpers of the cellular gatekeeper-miRNAs dance in P53 network. AB - Recent findings position microRNA (miRNAs) as novel key players in regulating expression of protein-coding genes by interfering with the stability and/or translation of specific target mRNAs and in controlling cell proliferation and differentiation by actively functioning in the p53 tumor suppression network. Accumulating evidence reveal the intimate relationship between p53 and miRNAs. P53 regulates the expression of miRNAs at multiple levels. miRNAs influence the post-transcriptional regulation and activities of p53 by invoking a myriad of mechanisms. These findings are unveiling the unknown mechanisms of the p53 mediated biological effects. PMID- 20006674 TI - Differential expression of exon 5 splice variants of sodium channel alpha subunit mRNAs in the developing mouse brain. AB - Sodium channel alpha subunit genes expressed in the human brain, SCN1A, SCN2A, SCN3A and SCN8A, are subject to alternative splicing of coding exons 5N and 5A. In this study we examined expression of alpha subunit mRNA and exon 5 splicing in the developing mouse brain. Expression levels of Scn1a, Scn2a and Scn8a mRNAs increase postnatally, whereas Scn3a mRNA expression levels decrease. Scn1a mRNA contains only exon 5A, due to the absence of exon 5N in the mouse Scn1a gene. At birth, Scn2a is the only sodium channel alpha subunit mRNA that contains higher or equal amounts of the 5N isoform compared to the 5A isoform in most brain regions. In contrast, the predominant isoform of Scn3a and Scn8a mRNAs in the newborn mouse brain is 5A. 5N/5A ratios for each of the three mRNAs vary across brain regions, with cortex >or= hippocampus>thalamus>cerebellum. In all brain regions and for all three alpha subunits, 5N/5A ratios gradually decrease with age, levelling at a value between 0.1 and 0.2. These findings suggest potential involvement of common factors in the alternative splicing of exon 5 for all three transcripts, and that expression of these factors varies between brain regions and changes during development. Differences in the strength of exon 5N and/or exon 5A splice sites in Scn2a pre-mRNA as compared to Scn1a and Scn8a may underlie the observed differences in 5N/5A ratios in the three alpha subunit mRNAs. PMID- 20006675 TI - Valproic acid reduces spatial working memory and cell proliferation in the hippocampus. AB - Valproic acid (VPA) is widely used clinically, as an anticonvulsant and mood stabilizer but is, however, also known to block cell proliferation through its ability to inhibit histone deacetylase enzymes. There have been a number of reports of cognitive impairments in patients taking VPA. In this investigation we examined the relationship between cognition and changes in cell proliferation within the hippocampus, a brain region where continued formation of new neurons is associated with learning and memory. Treatment of rats by i.p. injection of VPA, reduced cell proliferation in the sub granular zone of the dentate gyrus within the hippocampus. This was linked to a significant impairment in their ability to perform a hippocampus-dependent spatial memory test (novel object location). In addition, drug treatment caused a significant reduction in brain derived neurotrophic factor (BDNF) and Notch 1 but not doublecortin levels within the hippocampus. These results support the idea that VPA may cause cognitive impairment and provide a possible mechanism for this by reducing neurogenesis within the hippocampus. PMID- 20006676 TI - Protein kinase C deficiency-induced alcohol insensitivity and underlying cellular targets in Drosophila. AB - Multiple subtypes of protein kinase C (PKC) isozymes are implicated in various neurological disorders including alcohol insensitivity, a trait strongly associated with alcoholism in humans, but molecular and cellular mechanisms underlying the PKC activities remain poorly understood. Here we show that functional knockdown of conventional, novel or atypical PKC in the fly nervous system each resulted in alcohol insensitivity. Neuroanatomical mapping of conventional Ca(2+)-sensitive PKC53E activity uncovers a previously uncharacterized role of Drosophila serotonin neurons in alcohol sensitivity. The deficiency of PKC53E but not novel Ca(2+)-independent PKC98E appears to reduce synaptic serotonin levels, since acute inhibition of serotonin reuptake by citalopram and Prozac reversed alcohol insensitivity in flies expressing PKC53E double-stranded RNA in serotonin neurons. Together, findings from this and our previous studies indicate that PKC53E and PKC98E differentially regulate fly alcohol sensitivity through independent modulation of conserved serotonin and neuropeptide Y-like systems. PMID- 20006677 TI - Endogenous nociceptin/orphanin FQ (N/OFQ) contributes to haloperidol-induced changes of nigral amino acid transmission and parkinsonism: a combined microdialysis and behavioral study in naive and nociceptin/orphanin FQ receptor knockout mice. AB - The contribution of endogenous nociceptin/orphanin FQ (N/OFQ) to neuroleptic induced parkinsonism has been evaluated in haloperidol-treated mice. Pharmacological blockade of N/OFQ receptors (NOP) via systemic administration of 1-[(3R,4R)-1-cyclooctylmethyl-3-hydroxymethyl-4-piperidyl]-3-ethyl-1,3-dihydro-2H benzimidazol-2-one (J-113397, 0.01-10 mg/kg i.p.) or central injection of [Nphe(1),Arg(14),Lys(15)]N/OFQ-NH(2) (UFP-101, 10 nmol i.c.v.) attenuated (0.8 mg/kg) haloperidol-induced motor deficits as evaluated by a battery of behavioral tests providing complementary information on motor parameters: the bar, drag and rotarod tests. A combined neurochemical and behavioral approach was then used to investigate whether the substantia nigra reticulata could be involved in antiakinetic actions of J-113397. Microdialysis combined to the bar test revealed that haloperidol (0.3 and 0.8 mg/kg i.p.) caused a dose-dependent and prolonged elevation of immobility time (i.e. akinesia) which was associated with an increase in nigral glutamate and a reduction in GABA release. Conversely, J 113397 (1 mg/kg) alone reduced glutamate and elevated nigral GABA release, and when challenged against haloperidol, counteracted its behavioral and neurochemical effects. Microdialysis coupled to behavioral testing also demonstrated that NOP receptor knockout mice were resistant to haloperidol (0.3 mg/kg) compared to wild-type mice, lack of response being associated with a reversal of glutamate release facilitation into inhibition and no change in nigral GABA release. This study provides pharmacological and genetic evidence that endogenous N/OFQ contributes to haloperidol-induced akinesia and changes of amino acid transmission in mice. Moreover, it confirms the view that NOP receptor antagonists are capable of reversing akinesia across species and genotypes and may prove effective in relieving neuroleptic-induced parkinsonism. PMID- 20006678 TI - Conditioning lesions enhance growth state only in sensory neurons lacking calcitonin gene-related peptide and isolectin B4-binding. AB - A conditioning lesion improves regeneration of central and peripheral axons of dorsal root ganglion (DRG) neurons after a subsequent injury by enhancing intrinsic growth capacity. This enhanced growth state is also observed in cultured DRG neurons, which support a more sparsely and rapidly elongating mode of growth after a prior conditioning lesion in vivo. Here we examined differences in the capacity or requirements of specific types of sensory neurons for regenerative growth, which has important consequences for development of strategies to improve recovery after injury. We showed that after partial or complete injury of the sciatic nerve in mice, an elongating mode of growth in vitro was activated only in DRG neurons that did not express calcitonin gene related peptide (CGRP) or bind Bandeiraea simplicifolia I-isolectin B4 (IB4). We also directly examined the response of conditioned sensory neurons to nerve growth factor (NGF), which does not enhance growth in injured peripheral nerves in vivo. We showed that after partial injury, NGF stimulated a highly branched and linearly restricted rather than elongating mode of growth. After complete injury, the function of NGF was impaired, which immunohistochemical studies of DRG indicated was at least partly due to downregulation of the NGF receptor, tropomyosin-related kinase A (TrkA). These results suggest that, regardless of the type of conditioning lesion, each type of DRG neuron has a distinct intrinsic capacity or requirement for the activation of rapidly elongating growth, which does not appear to be influenced by NGF. PMID- 20006679 TI - The NG2 proteoglycan promotes oligodendrocyte progenitor proliferation and developmental myelination. AB - The NG2 proteoglycan has been shown to promote proliferation and motility in a variety of cell types. The presence of NG2 on oligodendrocyte progenitor cells (OPCs) suggests that the proteoglycan may be a factor in expansion of the OPC pool to fill the entire CNS prior to OPC differentiation to form myelinating oligodendrocytes. Comparisons of postnatal cerebellar myelination in wild type and NG2 null mice reveal reduced numbers of OPCs in developing white matter of the NG2 null mouse. Quantification of BrdU incorporation shows that reduced proliferation is a key reason for this OPC shortage, with the peak of OPC proliferation delayed by 4-5 days in the absence of NG2. As a result of the subnormal pool of OPCs, there is also a delay in production of mature oligodendrocytes and myelinating processes in the NG2 null cerebellum. NG2 may promote OPC proliferation via enhancement of growth factor signaling or mediation of OPC interaction with unmyelinated axons. PMID- 20006680 TI - Functional recovery of stepping in rats after a complete neonatal spinal cord transection is not due to regrowth across the lesion site. AB - Rats receiving a complete spinal cord transection (ST) at a neonatal stage spontaneously can recover significant stepping ability, whereas minimal recovery is attained in rats transected as adults. In addition, neonatally spinal cord transected rats trained to step more readily improve their locomotor ability. We hypothesized that recovery of stepping in rats receiving a complete spinal cord transection at postnatal day 5 (P5) is attributable to changes in the lumbosacral neural circuitry and not to regeneration of axons across the lesion. As expected, stepping performance measured by several kinematics parameters was significantly better in ST (at P5) trained (treadmill stepping for 8 weeks) than age-matched non-trained spinal rats. Anterograde tracing with biotinylated dextran amine showed an absence of labeling of corticospinal or rubrospinal tract axons below the transection. Retrograde tracing with Fast Blue from the spinal cord below the transection showed no labeled neurons in the somatosensory motor cortex of the hindlimb area, red nucleus, spinal vestibular nucleus, and medullary reticular nucleus. Retrograde labeling transsynaptically via injection of pseudorabies virus (Bartha) into the soleus and tibialis anterior muscles showed no labeling in the same brain nuclei. Furthermore, re-transection of the spinal cord at or rostral to the original transection did not affect stepping ability. Combined, these results clearly indicate that there was no regeneration across the lesion after a complete spinal cord transection in neonatal rats and suggest that this is an important model to understand the higher level of locomotor recovery in rats attributable to lumbosacral mechanisms after receiving a complete ST at a neonatal compared to an adult stage. PMID- 20006681 TI - Anorexia in rats caused by a valine-deficient diet is not ameliorated by systemic ghrelin treatment. AB - Rodents exhibit aversive behavior toward a diet that lacks at least one of the essential amino acids. We sought to determine whether the particular form of anorexia caused by such diets could be ameliorated by the administration of orexigenic peptides while simultaneously analyzing the neural mechanisms underlying anorexia. Rats were fed a valine-deficient diet, which induced severe anorexia (reducing food consumption by 80%). The severe anorexia was associated with a significant decrease in the cerebrospinal fluid valine concentration and hyper-ghrelinemia. Between 6 and 12 days after initiation of the valine-deficient diet, we injected rats twice daily with valine and/or an orexigenic peptide (ghrelin, neuropeptide Y, or agouti-related protein) either i.p. or i.c.v.. We then measured dietary intake. An i.c.v. valine injection allowed earlier food intake compared with an i.p valine injection and increased the density of c-Fos positive ependymal cells lining the third ventricle. Whereas an i.c.v. injection of ghrelin or neuropeptide Y increased consumption of the valine-deficient diet, i.p injection of ghrelin or i.c.v. injection of agouti-related protein did not. Following i.c.v. administration of either valine or ghrelin, we did not observe complete recovery of consumption of the valine-deficient diet. This may be due to the ineffectiveness of peripheral ghrelin and central agouti-related protein and/or to conditioned aversion to the valine-deficient diet. Since ghrelin is known to be involved in food anticipatory activities, whether the hyper ghrelinemia observed in valine-deficient rats play role in foraging behavior other than food intake is the future study to be investigated. PMID- 20006682 TI - Anterior intraparietal cortex codes complexity of observed hand movements. AB - Human and monkey studies clearly show that the anterior intraparietal area (AIP) is crucial for hand-related visuomotor transformations. Human AIP activates also during observation of hand actions, involving it in the mirror system. It is not known, however, whether its activation can also reflect a difference in the complexity of the observed action. In the present study we used functional magnetic resonance imaging (fMRI) to explore the activation of human area AIP during the observation of complex object-manipulation tasks (e.g. inserting a key in a lock and turning it) as compared to simple tasks (whole hand grasping of an object) executed with the left and the right hand in a first person perspective. The results show that, in general, both complex and simple tasks produced an activation of the fronto-parietal mirror system and that the activity of AIP in each hemisphere was higher during observation of the contralateral hand (hand identity effect). A Region-Of-Interest (ROI) analysis of the parietal activations responding to hand identity showed that each AIP was more active during the observation of complex with respect to simple tasks. In the right AIP this effect was stronger during observation of the contralateral hand, in the left AIP was strong during observation of both hands. This complexity-related property was not observed in the other activated areas. These findings support the concept that the observation of motor acts retrieves the internal representation of those same acts in the observer's motor system (direct-matching hypothesis based on the mirror neuron mechanism). PMID- 20006683 TI - When people matter more than money: an ERPs study. AB - In the present study, we showed that, in a social gambling task, individuals are influenced more by the type of social interaction than by the pattern of gains and losses. More precisely, the neural responses, as well as the level of pleasantness/unpleasantness following gains and losses, are modulated by social interaction factors. Here we present an Event-Related Potentials (ERPs) study in which three groups of participants were compared. Subjects were engaged in gambling tasks differing with regard to social factors: in a first condition, there was no social context; in a second condition, participants compared their outcomes with those of another individual; in a third condition, participants competed for a limited amount of money with another contender. In all conditions, all participants were revealed the outcome of an unselected alternative (non obtained outcome) prior to the payoff associated with their selected option (obtained outcome). In addition, affective ratings were measured after the outcomes were presented. In the group without social context, ERPs results replicated previous findings. Interestingly, the P200 was modulated by varying social contexts, suggesting that attentive resources allocated to payoffs in comparison and competitive situations are decreased presumably in favor of social cues. Furthermore, Feedback Related Negativity (FRN) was predictive of the subjective feeling of pleasantness/unpleasantness following monetary outcomes. The present data provide information about neural and cognitive processing underlying economic decision-making when other individuals are involved. PMID- 20006684 TI - A bioactive isoprenylated xanthone and other constituents of Garcinia edulis. AB - Phytochemical investigation on the root bark of Garcinia edulis (Clusiaceae) gave a new isoprenylated xanthone, 1,4,6-trihydroxy-3-methoxy-2-(3-methyl-2-butenyl)-5 (1,1-dimethyl-prop-2-enyl)xanthone (1), a known xanthone, forbexanthone (2) together with three known pentacyclic triterpenoids, friedelin, lupeol and lupeol acetate. The structure of the new compound was fully characterised by NMR spectroscopic analysis. Compound 1 showed significant anti-HIV-1 protease activity with IC(50) value of 11.3 microg/mL. Furthermore, compound 1 showed potent cytotoxic activity with LC(50) value of 2.36 microg/mL against brine shrimp larva in vitro. PMID- 20006685 TI - Sesquiterpene lactones from Sonchus arvensis L. and their antibacterial activity against Streptococcus mutans ATCC 25175. AB - Two new sesquiterpene lactones, 1beta-sulfate-5alpha, 6betaH-eudesma-3-en-12, 6alpha-olide (1) and 1beta-(p-hydroxyphenyl acetyl)-15-O-beta-D-glucopyranosyl 5alpha, 6betaH-eudesma-3-en-12, 6alpha-olide (2) were isolated from Sonchus arvensis L. (Asteraceae), together with eight known compounds. Their structures were elucidated through spectroscopic and chemical methods. They were evaluated for antibacterial activity. Among them, compounds 1 and 7 exhibited antibacterial activity against oral pathogen Streptococcus mutans ATCC 25175 with MIC values of 15.6 and 62.5 microg/ml, respectively. PMID- 20006687 TI - The virtues of ignorance. AB - Although ignorance and uncertainty are usually unwelcome feelings, they have unintuitive advantages for both human and non-human animals, which we review here. We begin with the perils of too much information: expertise and knowledge can come with illusions (and delusions) of knowing. We then describe how withholding information can counteract these perils: providing people with less information enables them to judge more precisely what they know and do not know, which in turn enhances long-term memory. Data are presented from a new experiment that illustrates how knowing what we do not know can result in helpful choices and enhanced learning. We conclude by showing that ignorance can be a virtue, as long as it is recognized and rectified. PMID- 20006686 TI - Domain is a moving target for relational learning. AB - The domain for relational learning was manipulated by varying the training set size for pigeons that had learned the same/different (S/D) concept. Six pigeons that had learned a S/D task with pairs of pictures with a set size of 1024 picture items had their training set size reduced to 8 items. Training on the reduced 8-item set was followed by transfer testing that was repeated four times. Transfer performance following reduction of the training set to 8 items was 9.2% less than it had been when the pigeons were trained with the 1024-item set, but 25.8% above chance. This partial abstract-concept learning remained constant over the four tests with novel stimuli. The results show that a broad domain established by a large expanding training set can once again become restricted by further training with a small training set. PMID- 20006688 TI - Benzene-derived N2-(4-hydroxyphenyl)-deoxyguanosine adduct: UvrABC incision and its conformation in DNA. AB - Benzene, a ubiquitous human carcinogen, forms DNA adducts through its metabolites such as p-benzoquinone (p-BQ) and hydroquinone (HQ). N(2)-(4-Hydroxyphenyl)-2' deoxyguanosine (N(2)-4-HOPh-dG) is the principal adduct identified in vivo by (32)P-postlabeling in cells or animals treated with p-BQ or HQ. To study its effect on repair specificity and replication fidelity, we recently synthesized defined oligonucleotides containing a site-specific adduct using phosphoramidite chemistry. We here report the repair of this adduct by Escherichia coli UvrABC complex, which performs the initial damage recognition and incision steps in the nucleotide excision repair (NER) pathway. We first showed that the p-BQ-treated plasmid was efficiently cleaved by the complex, indicating the formation of DNA lesions that are substrates for NER. Using a 40-mer substrate, we found that UvrABC incises the DNA strand containing N(2)-4-HOPh-dG in a dose- and time dependent manner. The specificity of such repair was also compared with that of DNA glycosylases and damage-specific endonucleases of E. coli, both of which were found to have no detectable activity toward N(2)-4-HOPh-dG. To understand why this adduct is specifically recognized and processed by UvrABC, molecular modeling studies were performed. Analysis of molecular dynamics trajectories showed that stable G:C-like hydrogen bonding patterns of all three Watson-Crick hydrogen bonds are present within the N(2)-4-HOPh-G:C base pair, with the hydroxyphenyl ring at an almost planar position. In addition, N(2)-4-HOPh-dG has a tendency to form more stable stacking interactions than a normal G in B-type DNA. These conformational properties may be critical in differential recognition of this adduct by specific repair enzymes. PMID- 20006689 TI - Enhanced glutathione biosynthetic capacity promotes resistance to As3+-induced apoptosis. AB - Trivalent arsenite (As(3+)) is a known human carcinogen capable of inducing both cellular transformation and apoptotic cell death by mechanisms involving the production of reactive oxygen species. The tripeptide antioxidant glutathione (GSH) constitutes a vital cellular defense mechanism against oxidative stress. While intracellular levels of GSH are an important determinant of cellular susceptibility to undergo apoptotic cell death, it is not known whether cellular GSH biosynthetic capacity per se regulates As(3+)-induced apoptosis. The rate limiting enzyme in GSH biosynthesis is glutamate cysteine ligase (GCL), a heterodimeric holoenzyme composed of a catalytic (GCLC) and a modifier (GCLM) subunit. To determine whether increased GSH biosynthetic capacity enhanced cellular resistance to As(3+)-induced apoptotic cell death, we utilized a mouse liver hepatoma (Hepa-1c1c7) cell line stably overexpressing both GCLC and GCLM. Overexpression of the GCL subunits increased GCL holoenzyme formation and activity and inhibited As(3+)-induced apoptosis. This cytoprotective effect was associated with a decrease in As(3+)-induced caspase activation, cleavage of caspase substrates and translocation of cytochrome c to the cytoplasm. In aggregate, these findings demonstrate that enhanced GSH biosynthetic capacity promotes resistance to As(3+)-induced apoptosis by preventing mitochondrial dysfunction and cytochrome c release and highlight the role of the GSH antioxidant defense system in dictating hepatocyte sensitivity to As(3+)-induced apoptotic cell death. PMID- 20006690 TI - Prostaglandin E1 encapsulated into lipid nanoparticles improves its anti inflammatory effect with low side-effect. AB - Prostaglandin E(1) (PGE1) shows various pharmacological activities including anti inflammation. However, the rapid metabolization and inactivation of the intravenously administered PGE1 during the first passage through the lungs result in significant non-compliance in clinical trials which greatly limits its application. The aim of this work was to prepare the lipid nanoparticles loading PGE1 to improve its anti-inflammatory effect with low side-effect. The experimental results showed that PGE1 loaded lipid nanoparticles (PLNs) could be successfully prepared by high pressure homogenization with particle size 68.1+/ 4.7 nm, zeta potential -3.32+/-0.37 mV and entrapment efficiency 92.1+/-1.3%. PLNs exhibited a sustained release with low burst drug release. PLNs could improve the inhibition effects of PGE1 on lipopolysaccharides (LPS)-induced TNF alpha expression on macrophage RAW264.7 cells, and improve the inhibition of lymphocyte to endothelial cell adhesion and ICAM-1 adhesion molecule expression on HUVEC and MDA-MB-468 cell membrane. No allergenicity, vascular and muscle irritation were induced in animals by PLNs even at double of the highest drug concentration of clinical infusion. As a result, PLNs could be a more potential delivery system for PGE1 in the treatment of inflammation-related diseases. PMID- 20006691 TI - Determination of surface heterogeneity of D-mannitol by sessile drop contact angle and finite concentration inverse gas chromatography. AB - The sensitivity of two techniques in tracking changes in surface energetics was investigated for a crystalline excipient, D-mannitol. Macroscopic crystals of D mannitol were grown from saturated water solution by slow cooling, and sessile drop contact angle was employed to measure the anisotropic surface energy. The facet-specific surface energy was consistent with localised hydroxyl group concentrations determined by X-ray photoelectron spectroscopy (XPS), and was also in excellent agreement with the surface energy distribution of the powder form of mannitol measured via a new methodology using inverse gas chromatography (IGC) at finite concentrations. The gamma(SV)(d) was found to vary between 39.5 mJ/m(2) and 44.1 mJ/m(2) for contact angle and between 40 mJ/m(2) and 49 mJ/m(2) for IGC measurements. We report here, a high level of surface heterogeneity on the native mannitol crystal surfaces. When the surfaces of both D-mannitol samples (powder and large single crystals) were modified by dichlorodimethylsilane to induce surface hydrophobicity, both IGC and contact angle revealed a homogeneous surface due to functionalisation of mannitol crystal surface with methyl groups resulting in gamma(SV)(d) of approximately 34 mJ/m(2). It was shown that both IGC and contact angle techniques are able to detect surface chemical variations and detailed surface energetic distribution. PMID- 20006692 TI - Possible reduction of effective thickness of intestinal unstirred water layer by particle drifting effect. AB - According to the present theory of oral absorption, in the case of solubility limited absorption, the absorbed amount would not increase despite an increase in dose or a decrease in particle size. However, many experimental observations suggested that the absorbed amount was often increased (though sub proportionally) as the dose strength increased. In addition, the particle size reduction was often effective to increase the absorbed amount even in the case of solubility limited absorption. Since an increase of the dose strength and a decrease of the particle size cause no or little change in solubility and the mean intestinal transit time, effective intestinal membrane permeability (P(eff)) should have changed. The previous theory postulated that drug particles do not exist in the unstirred water layer (UWL) which is adjacent to the intestinal membrane. However, many reports suggested that nano- to micro-scale drug particles existed in the UWL. In this case, the effective thickness of the UWL (h(eff)) could be smaller than the nominal thickness, resulting in an increase of P(eff). In the present study, h(eff) was simply calculated assuming that the reduction of h(eff) is in proportion to the surface area of drug particles in the UWL. When the particle drifting effect was taken into account, the discrepancy between the theoretical calculation and experimental observations was reduced. It was suggested that when the dose (mg)/particle diameter (microm) ratio exceeds 20, the particle drifting effect would become significant. PMID- 20006693 TI - Analysis of phase diagram and microstructural transitions in an ethyl oleate/water/Tween 80/Span 20 microemulsion system using high-resolution ultrasonic spectroscopy. AB - High-resolution ultrasonic spectroscopy was applied to analyse a pseudo-ternary phase diagram for a mixture consisting of water/ethyl oleate/Tween 80 and Span 20 at 25 degrees C. The measured changes in the ultrasonic velocity and attenuation with concentration of water in oil/surfactants mixtures showed several, well defined stages and transitions between them, which allowed construction of an 'ultrasonic' phase diagram. Quantitative analysis of the ultrasonic parameters enabled characterisation of various phases (microemulsion, liquid crystals and pseudo-bicontinuous) as well as evaluation of the state of the water and particle size in microemulsion phase. PMID- 20006694 TI - Two-tone auditory suppression in younger and older normal-hearing adults and its relationship to speech perception in noise. AB - One approach for establishing how age affects psychoacoustic abilities is to compare the performance of young and older adults with normal auditory sensitivity. The present study used this approach to determine if age affects two tone suppression - a reduction in masked thresholds (henceforth, unmasking) following the introduction of a second (suppressing) tone to a masker-plus-signal stimulus complex. A secondary goal of the study was to assess whether individual differences in suppression would predict identification scores for words presented in a forward masking noise. Unmasking was measured by comparing forward masked thresholds for a 2000 Hz signal with a tonal (2000 Hz) masker alone or the tonal masker plus a 2300 Hz tonal suppressor. Speech perception in noise was assessed by obtaining forward-masked speech reception thresholds (SRTs) for isolated words presented with speech-shaped noise. Young, but not older, normal hearing adults exhibited significant amounts of unmasking. Nineteen of the 20 young adults tested exhibited unmasking, whereas less that half of the 25 older participants exhibited any unmasking. The correlation between suppression as indexed by unmasking and SRTs in young adults was approximately -0.6, suggesting that more suppression was associated with lower SRTs. The findings suggest that auditory suppression is one of the few psychoacoustic abilities that demonstrate significant changes with age even for older adults with minimal hearing loss. PMID- 20006695 TI - Expression of prestin and Gata-3,-2,-1 mRNA in the rat organ of Corti during the postnatal period and in culture. AB - Based on observations that mutations of GATA-3 are responsible for the HDR syndrome (hypoparathyroidism, deafness, renal defects) and that GATA transcription factors have an important role to play in inner ear development, we hypothesized that these transcription factors may be involved in regulatory changes of prestin transcription. To prove this, we examined in parallel the expression of mRNA of prestin and Gata-3,-2 and Gata-1 in the organ of Corti during early postnatal development of rats and in organotypic cultures. Remarkable relations are observed between prestin and Gata-3,-2 expression in organ of Corti preparations in vivo and in vitro: (i) Gata-3,-2 expression display similar apical-basal gradients as prestin mRNA levels. (ii) The prestin expression increases between postnatal day two and postnatal day eight by a factor of about four in the apical and middle segments and by a factor of two in the basal part. Highly significant Pearson correlation coefficients were observed between Gata-3,-2 mRNA and prestin levels when the data were evaluated by regression analyses. (iii) Parallel changes of prestin mRNA and Gata-3,-2 mRNA levels were observed in response to thyroid hormone and to gemfibrozil application. These observations suggest a regulatory role played by the Gata-3,-2 transcription factors in prestin expression. PMID- 20006696 TI - Wound healing potential of methanolic extract of Trichosanthes dioica Roxb (fruits) in rats. AB - AIM OF THE STUDY: The present study provides a scientific evaluation for the wound healing potential of methanolic (MeOH) extract of TDR fruits. MATERIALS AND METHODS: Excision and incision wounds were inflicted upon three groups of six rats each. Group I was assigned as control (ointment base), Group II was treated with standard silver sulfadiazine (0.01%) cream. Group III was treated with 5% MeOH extract ointment. The parameters observed were percentage of wound contraction, epithelialization period, hydroxyproline content, tensile strength including histopathological studies. RESULTS: It was noted that the effect produced by the extract ointment showed significant (P<0.01) healing in both the wound models when compared with control group. All parameters such as wound contraction, epithelialization period, hydroxyproline content, tensile strength and histopathological studies showed significant changes when compared to control. CONCLUSION: The result shows that TDR extract ointment demonstrates wound healing potential in both excision and incision models. PMID- 20006697 TI - The importance of botellas and other plant mixtures in Dominican traditional medicine. AB - ETHNOPHARMACOLOGICAL RELEVANCE: Plant mixtures are understudied in ethnobotanical research. AIM OF THE STUDY: To investigate the importance of plant mixtures (remedies consisting of at least two plants) in Dominican traditional medicine. MATERIALS AND METHODS: A Spanish language questionnaire was administered to 174 Dominicans living in New York City (NYC) and 145 Dominicans living in the Dominican Republic (DR), including lay persons (who self-medicate with plants) and specialists (traditional healers). Plants were identified through specimens purchased in NYC botanica shops and Latino grocery shops, and from voucher collections. RESULTS: The percentage of mixtures as compared to single plants in plant use reports varied between 32 and 41%, depending on the geographic location (NYC or DR) and participant status (lay person or specialist). Respiratory conditions, reproductive health and genitourinary conditions were the main categories for which Dominicans use plant mixtures. Lay persons reported significantly more mixtures prepared as teas, mainly used in NYC to treat respiratory conditions. Specialists mentioned significantly more botellas (bottled herbal mixtures), used most frequently in the DR to treat reproductive health and genitourinary conditions. Cluster analysis demonstrated that different plant species are used to treat respiratory conditions as compared to reproductive health and genitourinary conditions. Interview participants believed that combining plants in mixtures increases their potency and versatility as medicines. CONCLUSIONS: The present study demonstrates the importance and complexity of plant mixtures in Dominican traditional medicine and the variation in its practices influenced by migration from the DR to NYC, shedding new light on the foundations of a particular ethnomedical system. PMID- 20006698 TI - Acrylamide catalytically inhibits topoisomerase II in V79 cells. AB - The vinyl monomer acrylamide is characterized by the presence of an alpha,beta unsaturated carbonyl group that makes it reactive towards thiol, hydroxyl or amino groups and towards the nucleophilic centers in DNA. The ability of acrylamide to chemically modify protein thiols has prompted us to consider topoisomerase II as one possible target of acrylamide, since agents targeting protein sulfhydryl groups act as either catalytic inhibitors or poisons of topoisomerase II. Nuclear extracts from V79 Chinese hamster cells incubated with acrylamide reduced topoisomerase II activity as inferred by an inability to convert kinetoplast DNA to the decatenated form. Nuclear extracts incubated with acrylamide pre-incubated with DTT converted kinetoplast DNA to the decatenated form, suggesting that acrylamide influences topoisomerase II activity through reaction with sulfhydryl groups on the enzyme. Furthermore, acrylamide did not induce the pBR322 DNA cleavage, as assessed by cleavage assay; thus, it cannot be regarded as a poison of topoisomerase II. As a catalytic inhibitor, acrylamide antagonizes the effect of etoposide, a topoisomerase II poison, as determined by clonogenic assay in V79 cells. This antagonism is confirmed by band depletion assay, from which it can be inferred that acrylamide reduces the level of catalytically active cellular topoisomerase II available for the action of etoposide. PMID- 20006699 TI - Transition from compensated hypertrophy to systolic heart failure in the spontaneously hypertensive rat: Structure, function, and transcript analysis. AB - Gene expression, determined by micro-array analysis, and left ventricular (LV) remodeling associated with the transition to systolic heart failure (HF) were examined in the spontaneously hypertensive rat (SHR). By combining transcript and gene set enrichment analysis (GSEA) of the LV with assessment of function and structure in age-matched SHR with and without HF, we aimed to better understand the molecular events underlying the onset of hypertensive HF. Failing hearts demonstrated depressed LV ejection fraction, systolic blood pressure, and LV papillary muscle force while LV end-diastolic and systolic volume and ventricular mass increased. 1431 transcripts were differentially expressed between failing and non-failing animals. GSEA identified multiple enriched gene sets, including those involving inflammation, oxidative stress, cell degradation and cell death, as well as TGF-beta and insulin signaling pathways. Our findings support the concept that these pathways and mechanisms may contribute to deterioration of cardiac function and remodeling associated with hypertensive HF. PMID- 20006700 TI - Spinally applied ketamine or morphine attenuate peripheral inflammation and hyperalgesia in acute and chronic phases of experimental arthritis. AB - Inflammation causes sensitization of peripheral and central nociceptive neurons. Pharmacological modulation of the latter has successfully been used for clinical pain relief. In particular, inhibitors of the NMDA glutamate receptor such as ketamine and agonists at the mu-opioid receptor such as morphine are broadly used. Besides driving the propagation of pain signals, spinal mechanisms are also discussed to modulate inflammation in the periphery. Here, we tested the hypothesis that intrathecally applied ketamine or morphine not only reduce pain related behavior, but also attenuate induction and maintenance of the inflammatory response in a model of chronic antigen-induced arthritis (AIA). Ketamine, morphine or vehicle was applied to the spinal cords of anesthesized animals with AIA. Swelling and histopathological changes were assessed after 6h (acute phase). Intrathecal catheters were implanted in another set of animals with AIA and substances were applied continuously. During the observation period of 21 days, inflammation and pain-related behavior were assessed. Ketamine and morphine significantly reduced arthritis severity as indicated by reduced joint swelling, but even more intriguingly by reduced infiltration with inflammatory cells and joint destruction in the acute and the chronic phase of arthritis. Morphine showed strong antinociceptive effects in the acute phase only, while the newly established effective dose for ketamine in a continuous application design reduced hyperalgesia in the acute and the chronic stage. In conclusion, both compounds exhibit anti-inflammatory effects during induction and maintenance of arthritis when applied intrathecally. These data thus propose a role of spinal NMDA- and opioid-receptors in the neuronal control of immune-mediated inflammation. PMID- 20006701 TI - Integrating (Q)SAR models, expert systems and read-across approaches for the prediction of developmental toxicity. AB - It has been estimated that reproductive and developmental toxicity tests will account for a significant proportion of the testing costs associated with REACH compliance. Consequently, the use of alternative methods to predict developmental toxicity is an attractive prospect. The present study evaluates a number of computational models and tools which can be used to aid assessment of developmental toxicity potential. The performance and limitations of traditional (quantitative) structure-activity relationship ((Q)SARs) modelling, structural alert-based expert system prediction and chemical profiling approaches are discussed. In addition, the use of category formation and read-across is also addressed. This study demonstrates the limited success of current modelling methods when used in isolation. However, the study also indicates that when used in combination, in a weight-of-evidence approach, better use may be made of the limited toxicity data available and predictivity improved. Recommendations are provided as to how this area could be further developed in the future. PMID- 20006702 TI - Stabilization of mitochondrial function by tetramethylpyrazine protects against kainate-induced oxidative lesions in the rat hippocampus. AB - Mitochondria are critical regulators of cell death, a key feature of neurodegeneration. Reactive oxygen species (ROS) are crucial to Ca(2+)-mediated effects of glutamate receptor activation leading to neuronal degeneration. Tetramethylpyrazine (TMP) is a principal ingredient of Ligusticum wallichi Franchat (a Chinese herb), used for treatment of cardiovascular and cerebrovascular ischemic diseases. However, its protection against oxidative brain injury associated with excessive activation of glutamate receptors is unknown. In this study, we demonstrate TMP neuroprotection against kainate induced excitotoxicity in vitro and in vivo. We found that TMP could partly alleviate kainate-induced status epilepticus in rats and prevented and rescued neuronal loss in the hippocampal CA3 but not the CA1 region. The partial prevention and rescue of neuronal loss by TMP were attributable to the preservation of the structural and functional integrity of mitochondria, evidenced by maintaining the mitochondrial membrane potential, ATP production, and complex I and III activities. Stabilization of mitochondrial function was linked to the observation that TMP could function as a reductant/antioxidant to quench ROS, block lipid peroxidation, and protect enzymatic antioxidants such as glutathione peroxidase and glutathione reductase. These results suggest that TMP may protect against oxidative brain injury by stabilization of mitochondrial function through quenching of ROS. PMID- 20006703 TI - Differential effects of chronic ethanol exposure on cytochrome P450 2E1 and the hypothalamic-pituitary-adrenal axis in the maternal-fetal unit of the guinea pig. AB - BACKGROUND: Ethanol neurobehavioural teratogenicity is a leading cause of developmental mental deficiency, in which the hippocampus is a target site of injury. The multi-faceted mechanism of ethanol teratogenicity is not completely understood. This study tested the hypothesis that chronic ethanol exposure (CEE), via chronic maternal ethanol administration, increases cytochrome P450 2E1 (CYP2E1) expression and alters hypothalamic-pituitary-adrenal (HPA) axis activity in the maternal-fetal unit during the third-trimester-equivalent of gestation. METHODS: Pregnant Dunkin-Hartley-strain guinea pigs received daily oral administration of ethanol (4 g ethanol/kg maternal body weight) or isocaloric sucrose/pair-feeding (control) throughout gestation (term, about gestational day (GD) 68). On GD 45, 55 and 65, pregnant animals were euthanized 2h after the last daily dose. Maternal and fetal body weights and fetal hippocampal brain weight were determined. Maternal and fetal samples were collected for the determination of liver CYP2E1 enzymatic activity and plasma free cortisol and ACTH concentrations. RESULTS: CEE, with maternal blood ethanol concentration of 108 124 mg/dl at 2h after the last dose, decreased fetal hippocampal weight only at GD 65 and had no effect on fetal body weight compared with control. CYP2E1 activity increased with gestational age in the fetal liver microsomal and mitochondrial fractions. CEE increased CYP2E1 activity in the microsomal and mitochondrial fractions of maternal liver at the three gestational ages and in both hepatic subcellular fractions of the GD 65 fetus compared with control. There was a gestational-age-dependent increase in maternal and fetal plasma free cortisol concentrations, but no effect of CEE compared with control. Maternal and fetal plasma ACTH concentrations were unaffected by CEE compared with control, and were virtually unchanged during the third-trimester-equivalent that was studied. CONCLUSION: These data demonstrate that, in the pregnant guinea pig, this CEE regimen increases liver CYP2E1 activity, without affecting HPA axis function, in the maternal-fetal unit during near-term gestation. The CEE-induced increase in liver CYP2E1 activity and potential oxidative stress in the maternal fetal unit may play a role in the pathogenesis of ethanol teratogenicity. PMID- 20006704 TI - Lead and cognitive function in VDR genotypes in the third National Health and Nutrition Examination Survey. AB - The relationship between the blood lead concentration and cognitive function in children and adults with different VDR genotypes who participated in the third National Health and Nutrition Examination Survey was investigated. The relationship between blood lead and serum homocysteine concentrations was also investigated. In children 12 to 16 years old, performance on the digit span and arithmetic tests as a function of the blood lead concentration varied by VDR rs2239185 and VDR rs731236 genotypes. Decreases in performance occurred in some genotypes, but not in others. In adults 20 to 59 years old, performance on the symbol-digit substitution test as a function of the blood lead concentration varied by VDR rs2239185-rs731236 haplotype. In the 12 to 16 year old children and adults 60 or more years old, the relationship between the serum homocysteine and blood lead concentrations varied by VDR genotype. The mean blood lead concentrations of the children and adults did not vary by VDR genotype. PMID- 20006705 TI - DeltaEF1 promotes breast cancer cell proliferation through down-regulating p21 expression. AB - Although the zinc finger-homeodomain transcription factor deltaEF1 is implied as a regulatory factor at the crossroad between proliferation and differentiation in carcinogenesis, its potential effect in the regulation of cell cycle progression has not been well elucidated. In our present study, we provide novel finding that, in breast cancer, the ectopic expression of deltaEF1 in MDA-MB-231 cells significantly promoted cell proliferation by increasing the cell number in S phase of the cell cycle. In contrast, deltaEF1 knockdown by RNA interference exhibited an opposite effect, highlighting a potent role of deltaEF1 to promote G1-S transition of breast cancer cells. Moreover, we demonstrated that deltaEF1 down-regulated p21 and concurrently up-regulated the expressions of CDK2 and CDK4 during this process. Further, deltaEF1 inhibited p21 transcription by recruiting to the E(2) box element on the p21 promoter. Depletion of endogenous deltaEF1 in MDA-MB-231 cells was sufficient to allow an inherent release of p21 expression, thus resulting in the cell cycle arrest. In addition, the stimulatory effect of deltaEF1 on cell proliferation through p21 regulation was supported by an inverse correlation of deltaEF1 and p21 expressions observed in both breast cancer cell lines and clinical tumor specimens. Taken together, these observations suggest a dual effect of deltaEF1 in promoting breast cancer cell proliferation, by differentially regulating the cell cycle regulatory proteins. PMID- 20006706 TI - The proximal STAT6 and NF-kappaB sites are responsible for IL-13- and TNF-alpha induced RhoA transcriptions in human bronchial smooth muscle cells. AB - RhoA protein is involved in the Ca(2+) sensitization of bronchial smooth muscle (BSM) contraction, and an upregulation of RhoA in BSMs has been suggested in allergic bronchial asthma. However, the mechanism of upregulation of RhoA remains poorly understood. In the present study, the transcriptional regulation of human RhoA gene was investigated in cultured human BSM cells stimulated with IL-13 and TNF-alpha, both of which have an ability to upregulate RhoA protein. Luciferase based assay showed that the RhoA promoter activity was augmented by both IL-13 and TNF-alpha. The deletion studies revealed a significant level of promoter activity between the 112 bp upstream and the transcription start site, which contains the STAT6 (78-70 bp upstream) and NF-kappaB (84-74 bp upstream) binding regions. The promoter activity was also decreased significantly by the mutations of these regions. Thus, the current study for the first time characterized the transcriptional regulation of the human RhoA gene. The findings also suggest that STAT6 and NF-kappaB are important for the upregulation of RhoA in human BSM induced by IL-13 and TNF-alpha, both of which are major cytokines in the pathogenesis of allergic bronchial asthma. PMID- 20006707 TI - Non-viral nanosystems for systemic siRNA delivery. AB - To use siRNA (small interfering ribonucleic acids) for systemic administration, a delivery system is often necessary to overcome barriers between administration and the target sites. These delivery systems require different properties to be efficient. On the one hand, they have to protect siRNA from degradation and/or inactivation and, on the other hand, they have themselves to be stable in blood and possess stealth properties to avoid elimination and degradation. Active and/or passive targeting should help the delivery system to reach the desired cell type or tissue, to be internalised, and to deliver siRNA to the cytoplasm so that siRNA can act by RNA interference and inhibit protein synthesis. This review presents an overview of different non-viral delivery systems, which have been evaluated in vivo or entered in clinical trials, with a focus on their physicochemical properties in order to help the development of new and efficient siRNA delivery systems, as the therapeutic solutions of tomorrow. PMID- 20006708 TI - Formate-nitrite transporters: optimisation of expression, purification and analysis of prokaryotic and eukaryotic representatives. AB - The formate-nitrite transporter family is composed of integral membrane proteins that possess six to eight alpha-helical transmembrane domains. Genes encoding these proteins are observed widely in prokaryotic genomes as well as certain groups of lower eukaryotes. Thus far, no structural information is available for this transporter family. Towards this aim, and to provide protein for biophysical studies, overexpression of a prokaryotic (TpNirC, from the hyperthermophilic archaebacterium Thermofilum pendens) and an eukaryotic (AnNitA, from the fungus Aspergillus nidulans) representative was achieved in Escherichia coli and Pichia pastoris hosts, respectively. The proteins were purified to >95% homogeneity yielding quantities sufficient for crystallisation trials and were shown by Circular Dichroism (CD) spectroscopy to have a highly alpha-helical content as expected from in silico predictions. Preliminary investigations by size exclusion chromatography of the oligomeric state of the purified AnNitA protein suggested that it most likely exists as a tetramer. PMID- 20006709 TI - Expression of soluble and functional human neonatal Fc receptor in Pichia pastoris. AB - The neonatal Fc receptor (FcRn) is a non-covalently associated heterodimeric protein composed of a transmembrane anchored heavy chain (alphaFcRn) and a soluble light chain beta2-microglobulin (beta2m). In addition to its role in the transfer of maternal immunoglobulin Gs (IgGs) to the fetus, FcRn plays a key role in prolonging the serum half-life of IgGs in vivo. Herein, we report a strategy for functional expression of soluble human FcRn (shFcRn) in Pichia pastoris using a two-promoter vector system, where alphaFcRn and beta2m are co-expressed under their respective promoters in a single vector. The purified shFcRn from the culture supernatants correctly assembled to form the heterodimer with the typical secondary structures. At acidic pHs between 5.0 and 6.4, shFcRn exhibited substantial binding to the four subclasses of human IgGs at acidic pHs between 5.0 and 6.4, but at pHs between 6.8 and 8.0, its binding was negligible binding. No cross-reactivity with mouse IgG was exhibited even at acidic pH. This was consistent with the pH-dependent binding profiles of the shFcRn prepared from the mammalian cell expression. Furthermore, the shFcRn exhibited about 10-fold higher binding affinity with the tumor necrosis factor-alpha antagonists of monoclonal antibodies Infliximab and Adalimumab than that of Etanercept, providing a clue to their different serum half-lives in vivo. Our results suggest that the functionally expressed shFcRn from Pichia can be used for the biochemical and biological studies and as a screening probe for Fc engineering of human IgGs. PMID- 20006710 TI - Analysis of the pyramidal tract in tumor patients using diffusion tensor imaging. AB - In this work, we propose to use fiber tracking in order to analyze and quantify the state of the pyramidal tracts in patients affected by tumors. We introduce a framework that includes an automatic method to obtain the fibers involved in the pyramidal tract of any subject, in order to compare robustly fiber bundles affected by tumors with healthy fiber tracts from control subjects and also to quantify the relative state of degeneration between the fiber tracts in the two hemispheres of the same patient. The comparative analyses proposed in our methodology are based on a new set of measures on the pyramidal tract, which take into account intrinsic properties of the fibers involved in the bundle as well as the similarity with the pyramidal tract of a standard healthy subject, modeled as the average of a set of controls. In order to perform better comparison studies and to take into account more information of the whole bundle, a mapping technique is used to represent the fiber tracts in 2D. Here, we show a set of experiments using 5 tumor patients and 10 control subjects, including pre- and post-operative studies in patients that have been treated with partial or total tumor resection. The results obtained indicate the usefulness of the method showing good overall performance. A reproducibility study using several acquisitions of the same patient is also presented to validate the techniques employed. PMID- 20006711 TI - Hierarchical control of false discovery rate for phase locking measures of EEG synchrony. AB - Computing phase-locking values (PLVs) between EEG signals is becoming a popular measure for quantifying functional connectivity, because it affords a more detailed picture of the synchrony relationships between channels at different times and frequencies. However, the accompanying increase in data dimensionality incurs a serious multiple testing problem for determining PLV significance. Standard methods for controlling Type I error, which treat all hypotheses as belonging to a single family, can fail to detect any significant discoveries. Instead, we propose a novel application of a hierarchical FDR method, which subsumes multiple families, for detecting significant PLV effects. For simulations and experimental data, we show that the proposed hierarchical FDR method is most powerful. This method revealed significant synchrony effects in the expected regions at an acceptable error rate of 5%, where other methods, including standard FDR correction failed to reveal any significant effects. PMID- 20006712 TI - Avoiding non-independence in fMRI data analysis: leave one subject out. AB - Concerns regarding certain fMRI data analysis practices have recently evoked lively debate. The principal concern regards the issue of non-independence, in which an initial statistical test is followed by further non-independent statistical tests. In this report, we propose a simple, practical solution to reduce bias in secondary tests due to non-independence using a leave-one-subject out (LOSO) approach. We provide examples of this method, show how it reduces effect size inflation, and suggest that it can serve as a functional localizer when within-subject methods are impractical. PMID- 20006713 TI - Impairment and compensation coexist in amnestic MCI default mode network. AB - Mild cognitive impairment (MCI) is the transitional, heterogeneous continuum from healthy elderly to Alzheimer's disease (AD). Previous studies have shown that brain functional activity in the default mode network (DMN) is impaired in AD patients. However, altering DMN activity patterns in MCI patients remains largely unclear. The present study utilized resting-state functional magnetic resonance imaging (fMRI) and an independent component analysis (ICA) approach to investigate DMN activity in 14 amnestic MCI (aMCI) patients and 14 healthy elderly. Compared to the aMCI patients, the healthy elderly exhibited increased functional activity in the DMN regions, including the bilateral precuneus/posterior cingulate cortex, right inferior parietal lobule, and left fusiform gyrus, as well as a trend towards increased right medial temporal lobe activity. The aMCI patients exhibited increased activity in the left prefrontal cortex, inferior parietal lobule, and middle temporal gyrus compared to the healthy elderly. Increased frontal-parietal activity may indicate compensatory processes in the aMCI patients. These findings suggest that abnormal DMN activity could be useful as an imaging-based biomarker for the diagnosis and monitoring of aMCI patients. PMID- 20006714 TI - Functional connectivity: shrinkage estimation and randomization test. AB - We develop new statistical methods for estimating functional connectivity between components of a multivariate time series and for testing differences in functional connectivity across experimental conditions. Here, we characterize functional connectivity by partial coherence, which identifies the frequency band (or bands) that drives the direct linear association between any pair of components of a multivariate time series after removing the linear effects of the other components. Partial coherence can be efficiently estimated using the inverse of the spectral density matrix. However, when the number of components is large and the components of the multivariate time series are highly correlated, the spectral density matrix estimate may be numerically unstable and consequently gives partial coherence estimates that are highly variable. To address the problem of numerical instability, we propose a shrinkage-based estimator which is a weighted average of a smoothed periodogram estimator and a scaled identity matrix with frequency-specific weight computed objectively so that the resulting shrinkage estimator minimizes the mean-squared error criterion. Compared to typical smoothing-based estimators, the shrinkage estimator is more computationally stable and gives a lower mean squared error. In addition, we develop a randomization method for testing differences in functional connectivity networks between experimental conditions. Finally, we report results from numerical experiments and analyze an EEG data set recorded during a visually guided hand movement task. PMID- 20006715 TI - Cortical thickness or grey matter volume? The importance of selecting the phenotype for imaging genetics studies. AB - Choosing the appropriate neuroimaging phenotype is critical to successfully identify genes that influence brain structure or function. While neuroimaging methods provide numerous potential phenotypes, their role for imaging genetics studies is unclear. Here we examine the relationship between brain volume, grey matter volume, cortical thickness and surface area, from a genetic standpoint. Four hundred and eighty-six individuals from randomly ascertained extended pedigrees with high-quality T1-weighted neuroanatomic MRI images participated in the study. Surface-based and voxel-based representations of brain structure were derived, using automated methods, and these measurements were analysed using a variance-components method to identify the heritability of these traits and their genetic correlations. All neuroanatomic traits were significantly influenced by genetic factors. Cortical thickness and surface area measurements were found to be genetically and phenotypically independent. While both thickness and area influenced volume measurements of cortical grey matter, volume was more closely related to surface area than cortical thickness. This trend was observed for both the volume-based and surface-based techniques. The results suggest that surface area and cortical thickness measurements should be considered separately and preferred over gray matter volumes for imaging genetic studies. PMID- 20006717 TI - Altered activation in association with reward-related trial-and-error learning in patients with schizophrenia. AB - In patients with schizophrenia, the ability to learn from reinforcement is known to be impaired. The present fMRI study aimed at investigating the neural correlates of reinforcement-related trial-and-error learning in 19 schizophrenia patients and 20 healthy volunteers. A modified gambling paradigm was applied where each cue indicated a subsequent number which had to be guessed. In order to vary predictability, the cue-number associations were based on different probabilities (50%, 81%, 100%) which the participants were not informed about. Patients' ability to learn contingencies on the basis of feedback and reward was significantly impaired. While in healthy volunteers increasing predictability was associated with decreasing activation in a fronto-parietal network, this decrease was not detectable in patients. Analysis of expectancy-related reinforcement processing yielded a hypoactivation in putamen, dorsal cingulate and superior frontal cortex in patients relative to controls. Present results indicate that both reinforcement-associated processing and reinforcement learning might be impaired in the context of the disorder. They moreover suggest that the activation deficits which patients exhibit in association with the processing of reinforcement might constitute the basis for the learning deficits and their accompanying activation alterations. PMID- 20006716 TI - Time-frequency dynamics of resting-state brain connectivity measured with fMRI. AB - Most studies of resting-state functional connectivity using fMRI employ methods that assume temporal stationarity, such as correlation and data-driven decompositions computed across the duration of the scan. However, evidence from both task-based fMRI studies and animal electrophysiology suggests that functional connectivity may exhibit dynamic changes within time scales of seconds to minutes. In the present study, we investigated the dynamic behavior of resting state connectivity across the course of a single scan, performing a time frequency coherence analysis based on the wavelet transform. We focused on the connectivity of the posterior cingulate cortex (PCC), a primary node of the default-mode network, examining its relationship with both the "anticorrelated" ("task-positive") network as well as other nodes of the default-mode network. It was observed that coherence and phase between the PCC and the anticorrelated network was variable in time and frequency, and statistical testing based on Monte Carlo simulations revealed the presence of significant scale-dependent temporal variability. In addition, a sliding-window correlation procedure identified other regions across the brain that exhibited variable connectivity with the PCC across the scan, which included areas previously implicated in attention and salience processing. Although it is unclear whether the observed coherence and phase variability can be attributed to residual noise or modulation of cognitive state, the present results illustrate that resting-state functional connectivity is not static, and it may therefore prove valuable to consider measures of variability, in addition to average quantities, when characterizing resting-state networks. PMID- 20006718 TI - Functional source separation improves the quality of single trial visual evoked potentials recorded during concurrent EEG-fMRI. AB - EEG quality is a crucial issue when acquiring combined EEG-fMRI data, particularly when the focus is on using single trial (ST) variability to integrate the data sets. The most common method for improving EEG data quality following removal of gross MRI artefacts is independent component analysis (ICA), a completely blind source separation technique. In the current study, a different approach is proposed based on the functional source separation (FSS) algorithm. FSS is an extension of ICA that incorporates prior knowledge about the signal of interest into the data decomposition. Since in general the part of the EEG signal that will contain the most relevant information is known beforehand (i.e. evoked potential peaks, spectral bands), FSS separates the signal of interest by exploiting this prior knowledge without renouncing the advantages of using only information contained in the original signal waveforms. A reversing checkerboard stimulus was used to generate visual evoked potentials (VEPs) in healthy control subjects. Gradient and ballistocardiogram artefacts were removed with template subtraction techniques to form the raw data, which were then subjected to ICA denoising and FSS. The resulting EEG data sets were compared using several metrics derived from average and ST data and correlated with fMRI data. In all cases, ICA was an improvement on the raw data, but the most obvious improvement was provided by FSS, which consistently outperformed ICA. The results show the benefit of FSS for the recovery of good quality single trial evoked potentials during concurrent EEG-fMRI recordings. PMID- 20006719 TI - Acute moderate exercise elicits increased dorsolateral prefrontal activation and improves cognitive performance with Stroop test. AB - A growing number of human studies have reported the beneficial influences of acute as well as chronic exercise on cognitive functions. However, neuroimaging investigations into the neural substrates of the effects of acute exercise have yet to be performed. Using multichannel functional near-infrared spectroscopy (fNIRS), we sought cortical activation related to changes in the Stroop interference test, elicited by an acute bout of moderate exercise, in healthy volunteers (N=20). The compactness and portability of fNIRS allowed on-site cortical examination in a laboratory with a cycle ergometer, enabling strict control of the exercise intensity of each subject by assessing their peak oxygen intake (VO2peak). We defined moderate exercise intensity as 50% of a subject's peak oxygen uptake (50%VO2peak). An acute bout of moderate exercise caused significant improvement of cognitive performance reflecting Stroop interference as measured by reaction time. Consistent with previous functional neuroimaging studies, we detected brain activation due to Stroop interference (incongruent minus neutral) in the lateral prefrontal cortices in both hemispheres. This Stroop-interference-related activation was significantly enhanced in the left dorsolateral prefrontal cortex due to the acute bout of moderate exercise. The enhanced activation significantly coincided with the improved cognitive performance. This suggests that the left dorsolateral prefrontal cortex is likely the neural substrate for the improved Stroop performance elicited by an acute bout of moderate exercise. fNIRS, which allows physiological monitoring and functional neuroimaging to be combined, proved to be an effective tool for examining the cognitive effects of exercise. PMID- 20006720 TI - Dynamic subcortical blood flow during male sexual activity with ecological validity: a perfusion fMRI study. AB - This study used arterial spin labeling (ASL) fMRI to measure brain perfusion in a group of healthy men under conditions that closely resembled customary sexual behavior. Serial perfusion measures for 30 min during two self-limited periods of partnered penis stimulation, and during post-stimulatory periods, revealed novel sexual activity-related cerebral blood flow (rCBF) changes, mainly in subcortical parts of the brain. Ventral pallidum rCBF was highest during the onset of penile erection, and lowest after the termination of penis stimulation. The perceived level of sexual arousal showed the strongest positive association with rCBF in the right basal forebrain. In addition, our results demonstrate that distinct subregions of the hypothalamus and cingulate cortex subserve opposite functions during human male sexual behavior. The lateral hypothalamus and anterior part of the middle cingulate cortex showed increased rCBF correlated with penile erection. By contrast, the anteroventral hypothalamus and subgenual anterior cingulate cortex exhibited rCBF changes correlated with penile detumescence after penile stimulation. Continuous rapid and high-resolution brain perfusion imaging during normal sexual activity has provided novel insights into the central mechanisms that control male sexual arousal. PMID- 20006721 TI - Propagation of epileptic spikes reconstructed from spatiotemporal magnetoencephalographic and electroencephalographic source analysis. AB - The purpose of this study is to assess the accuracy of spatiotemporal source analysis of magnetoencephalography (MEG) and scalp electroencephalography (EEG) for representing the propagation of frontotemporal spikes in patients with partial epilepsy. This study focuses on frontotemporal spikes, which are typically characterized by a preceding anterior temporal peak followed by an ipsilateral inferior frontal peak. Ten patients with frontotemporal spikes on MEG/EEG were studied. We analyzed the propagation of temporal to frontal epileptic spikes on both MEG and EEG independently by using a cortically constrained minimum norm estimate (MNE). Spatiotemporal source distribution of each spike was obtained on the cortical surface derived from the patient's MRI. All patients underwent an extraoperative intracranial EEG (IEEG) recording covering temporal and frontal lobes after presurgical evaluation. We extracted source waveforms of MEG and EEG from the source distribution of interictal spikes at the sites corresponding to the location of intracranial electrodes. The time differences of the ipsilateral temporal and frontal peaks as obtained by MEG, EEG and IEEG were statistically compared in each patient. In all patients, MEG and IEEG showed similar time differences between temporal and frontal peaks. The time differences of EEG spikes were significantly smaller than those of IEEG in nine of ten patients. Spatiotemporal analysis of MEG spikes models the time course of frontotemporal spikes as observed on IEEG more adequately than EEG in our patients. Spatiotemporal source analysis may be useful for planning epilepsy surgery, by predicting the pattern of IEEG spikes. PMID- 20006722 TI - Conservation phylogenetics of helodermatid lizards using multiple molecular markers and a supertree approach. AB - We analyzed both mitochondrial (mt-) and nuclear (n) DNAs in a conservation phylogenetic framework to examine deep and shallow histories of the Beaded Lizard (Heloderma horridum) and Gila Monster (H. suspectum) throughout their geographic ranges in North and Central America. Both mtDNA and intron markers clearly partitioned each species. One intron and mtDNA further subdivided H. horridum into its four recognized subspecies (H. n. alvarezi, charlesbogerti,exasperatum, and horridum). However, the two subspecies of H. suspectum (H. s. suspectum and H. s. cinctum) were undefined. A supertree approach sustained these relationships. Overall, the Helodermatidae is reaffirmed as an ancient and conserved group. Its most recent common ancestor (MRCA) was Lower Eocene [35.4 million years ago (mya)], with a approximately 25 my period of stasis before the MRCA of H. horridum diversified in Lower Miocene. Another approximately 5 my passed before H. h. exasperatum and H. h. horridum diverged, followed by approximately 1.5 my before H. h. alvarezi and H. h. charlesbogerti separated. Heloderma suspectum reflects an even longer period of stasis (approximately 30 my) before diversifying from its MRCA. Both H. suspectum (Mexico) and H. h. alvarezi also revealed evidence of historic range expansion following a recent bottleneck. Our conservation phylogenetic approach emphasizes the origin and diversification of this group, yields information on the manner by which past environmental variance may have impacted its populations and, in turn, allows us to disentangle historic from contemporary impacts that might threaten its long term persistence. The value of helodermatid conservation resides in natural services and medicinal products, particularly venom constituents, and these are only now being realized. PMID- 20006723 TI - Mitochondrial DNA evidence for deep genetic divergences in allopatric populations of the rocky intertidal isopod Ligia occidentalis from the eastern Pacific. AB - Nucleotide sequences from the cytochrome c oxidase subunit I (COI) gene were used to test for genetic differentiation in the rocky intertidal isopod crustacean, Ligia occidentalis (Ligiidae), from the eastern Pacific. Phylogenetic analyses showed that individuals of L. occidentalis from southern California, USA to Manzanillo, Colima, Mexico partitioned into 15 highly-divergent clades. Mean Kimura 2-parameter (K2P) genetic distances among clades ranged from 13.2% to 26.7%. These values are similar to interspecific genetic distances found in a wide variety of crustaceans, including Ligia spp., suggesting that the taxon L. occidentalis represents a complex of cryptic species. PMID- 20006724 TI - Phylogenetic analysis of European Scutovertex mites (Acari, Oribatida, Scutoverticidae) reveals paraphyly and cryptic diversity: A molecular genetic and morphological approach. AB - The soil and moss dwelling oribatid mite family Scutoverticidae is considered to represent an assemblage of distantly related but morphologically similar genera. We used nucleotide sequences of one mitochondrial (COI) and two nuclear (28S rDNA, ef-1alpha) genes, and 79 morphological characters to elucidate the phylogenetic relationships among 11 nominal plus two undescribed European mite species of the family Scutoverticidae with a particular focus on the genus Scutovertex. Both molecular genetic and morphological data revealed a paraphyletic genus Scutovertex, with S. pictus probably representing a distinct genus, and Provertex kuehnelti was confirmed as member of the family Scutoverticidae. Molecular genetic data confirmed several recently described Scutovertex species and thus the high species diversity within this genus in Europe and suggest that S. sculptus represents a complex of several cryptic species exhibiting marked genetic, but hardly any morphological divergence. PMID- 20006725 TI - Methodological considerations for improving Western blot analysis. AB - The need for a technique that could allow the determination of antigen specificity of antisera led to the development of a method that allowed the production of a replica of proteins, which had been separated electrophoretically on polyacrylamide gels, on to a nitrocellulose membrane. This method was coined Western blotting and is very useful to study the presence, relative abundance, relative molecular mass, post-translational modification, and interaction of specific proteins. As a result it is utilized routinely in many fields of scientific research such as chemistry, biology and biomedical sciences. This review serves to touch on some of the methodological conditions that should be considered to improve Western blot analysis, particularly as a guide for graduate students but also scientists who wish to continue adapting this now fundamental research tool. PMID- 20006726 TI - A unifying mechanism accounts for sensing of membrane curvature by BAR domains, amphipathic helices and membrane-anchored proteins. AB - The discovery of proteins that recognize membrane curvature created a paradigm shift by suggesting that membrane shape may act as a cue for protein localization that is independent of lipid or protein composition. Here we review recent data on membrane curvature sensing by three structurally unrelated motifs: BAR domains, amphipathic helices and membrane-anchored proteins. We discuss the conclusion that the curvature of the BAR dimer is not responsible for sensing and that the sensing properties of all three motifs can be rationalized by the physicochemical properties of the curved membrane itself. We thus anticipate that membrane curvature will promote the redistribution of proteins that are anchored in membranes through any type of hydrophobic moiety, a thesis that broadens tremendously the implications of membrane curvature for protein sorting, trafficking and signaling in cell biology. PMID- 20006727 TI - Mitochondrial dynamics in model organisms: what yeasts, worms and flies have taught us about fusion and fission of mitochondria. AB - Mitochondrial fusion and fission are important for a great variety of cellular functions, including energy metabolism, development, aging and cell death. Many of the core components mediating mitochondrial dynamics in human cells have been first identified and mechanistically analyzed in model organisms, such as Saccharomyces cerevisiae, Caenorhabditis elegans and Drosophila melanogaster. In particular, the functions of FZO/mitofusin and Mgm1/EAT-3/OPA1 in fusion and Dnm1/DRP1 in fission have been remarkably well conserved in yeasts, worms, flies and mammals. On the other hand, mechanisms to coordinate and regulate the activity of these molecular machines appear to be more diverse in different organisms. Here, I will discuss how S. cerevisiae, C. elegans and Drosophila have contributed to our current understanding of the cellular machineries mediating the dynamic behaviour of mitochondria. PMID- 20006729 TI - Lymphoid enhancer factor-1 mediates loading of Pax3 to a promoter harbouring lymphoid enhancer factor-1 binding sites resulting in enhancement of transcription. AB - The transcription factor, Pax3, alters transcription by binding directly to promoter regions harbouring sequences recognized by either its paired domain or its homeodomain. We demonstrated previously that the promoter regions of many of the genes whose expression was altered during a Pax3-induced mesenchymal-to epithelial transition harboured sequences recognized by lymphoid enhancer factor 1 (Lef1). Given the apparent lack of DNA-binding consensus sequences for Pax3 in these promoters, it was hypothesized that Pax3 might alter transcriptional activity of promoters harbouring Lef1-binding sites independent of Pax3 binding to DNA. We describe here a novel mode of Pax3-dependent regulation of transcription that is mediated through DNA-independent binding to Lef1. Specifically, we demonstrate that Pax3 binds to Lef1, determined in binding assays and co-immunoprecipitation of endogenous Pax3 and Lef1. Binding assays employing deletion mutants of Pax3 and Lef1 determined that association was mediated through the homeodomain of Pax3 and the first half of the Lef1 DNA binding domain. The significance of this association was demonstrated in transcriptional assays using a luciferase reporter gene downstream of a model promoter harbouring Lef1 DNA-binding consensus sites. Pax3 augmented Lef1 dependent transactivation from this promoter. This increase in transcriptional activity occurred in the absence and presence of added beta-catenin. Chromatin immunoprecipitation assays demonstrated further that Pax3-association to complexes bound to DNA harbouring Lef1 consensus sequences was dependent on Lef1. These data reveal a novel mode of transcriptional regulation by Pax3. This mode of transcriptional regulation suggests further that Pax3 activity may directly effect the expression of factors regulated by signal transduction pathways dependent on Lef1. PMID- 20006728 TI - Development and use of the lens epithelial explant system to study lens differentiation and cataractogenesis. AB - Over the last two decades much progress has been made in identifying and characterizing many of the molecules involved in understanding normal lens biology and its pathology. Much of this has been made possible through the establishment and use of the lens epithelial explant system. This simplistic tissue culture model, comprised of a sheet of lens epithelium on its native substratum, has been used effectively to study many cellular processes, including lens epithelial cell proliferation, fiber cell differentiation, cell apoptosis as well as epithelial-to-mesenchymal transformation of cells. In doing so, a number of key growth factors and cytokines, including members of the FGF, Wnt and TGFbeta family have been shown to play essential roles in many of these cellular events. This has led to further studies exploring the signaling pathways downstream of these molecules in the lens, paving the way for the development of a number of in situ models (primarily transgenic mouse lines) to further explore in more detail the nature of these molecular and cellular interactions. To reciprocate, the lens epithelial explant system is increasingly being used to further characterize the nature of many complex phenotypes and pathologies observed in these in situ models, allowing us to selectively isolate and examine the direct impact of an individual molecule on a specific cellular response in lens cells. There is no question that the lens epithelial explant system has served as a powerful tool to further our understanding of lens biology and pathology, and there is no doubt that it will continue to serve in such a capacity, as new developments are realized and putative treatments for aberrant lens cell behavior are to be trialed. PMID- 20006730 TI - Neuropathy target esterase: an essential enzyme for neural development and axonal maintenance. AB - Neuropathy target esterase (NTE) is an endoplasmic reticulum-anchored protein conserved across species. The N-terminal regulatory region of NTE contains three cyclic nucleotide binding domains while the C-terminal catalytic domain has a patatin domain. The NTE gene is expressed in mouse early at embryonic day 7 and its expression is maintained throughout embryonic development. NTE protein is mainly distributed in the nervous system with a pattern that is more restricted to large neurons in older animals. NTE regulates phospholipid metabolism and is known to be a phospholipase B. Knockout of NTE is embryonic lethal in mice, indicating that NTE is essential for embryonic survival. Neuronal specific NTE knockouts survive to adulthood, but show vacuolation and neuronal loss characteristic of neurodegenerative diseases. Recently, mutations in human NTE have been shown to cause a hereditary spastic paraplegia called NTE-related motor neuron disorder, suggesting a critical role for NTE in the nervous system. PMID- 20006731 TI - Phenolic fraction of tobacco smoke condensate potentiates benzo[a]pyerene diol epoxide-induced cell transformation: role of protein kinase C. AB - In this study we separated weakly acidic phenolic components from other neutral, acidic and basic components of tobacco smoke condensate (TSC) and observed that phenolic fraction of TSC significantly increased the number of colonies of promotion-sensitive JB6 Cl41 cells that showed anchorage-independent growth on soft agar in response to BPDE (an ultimate carcinogen produced by metabolic activation of the PAH benzo[a]pyrene). Anchorage-independent cell growth is indicative of cell transformation resulting in acquisition of tumorigenic potential. In order to understand the underlying mechanism by which TSC phenolic fraction potentiates BPDE-induced tumorigenicity, we examined its effect on the activation of two transcription factors AP-1 and NF-kappaB which are known to be influenced by established tumor promoter TPA. BPDE treatment caused induction of both AP-1 and NF-kappaB activity as determined by luciferase reporter assay and only NF-kappaB induction in response to BPDE was significantly attenuated by TSC phenolic fraction whereas AP-1 induction remains unaltered. Attenuation of NF kappaB activation by TSC phenolic fraction was associated with significant decrease of intracellular PKC substrate phosphorylation in BPDE treated cells. Non-specific PKC inhibitors staurosporine and bisindolylmaleimide II as well as inhibitors specific to conventional PKCs (Go6976) and PKC-delta (rottlerin) attenuated NF-kappaB activation in BPDE treated cells to a varying degree indicating a possible link between PKC down-regulation and the attenuation of NF kappaB activity by TSC phenolic fraction. Treatment of cells with PKC inhibitors also potentiated anchorage-independent growth of BPDE treated cells on soft agar. Our data suggest a possible role of PKC down-regulation in potentiation of BPDE induced tumorogenicity by TSC phenolic fraction. PMID- 20006732 TI - Aneugenic effects of benzoxazinones in cultured human cells. AB - Benzoxazinones (BAs) are natural products that are present in Gramineae and represent part of the plant defence system against pests. In recent years, sprouts of maize, wheat and rye have been used for the production of dietary supplements. We have investigated the potential genotoxic activities of a diverse range of synthetic derivatives of the most abundant natural BA, namely DIBOA (2,4 dihydroxy-1,4-benzoxazin-3-one), proposed for use as a potential herbicide. We have tested 18 synthetic BAs for potential effects in cultured HeLa cells. We found significantly higher micronucleus (MN) induction over the background level, with the solvent DMSO used as an internal control. Concentration-dependent effects were found between 1nM and 20nM for all the synthetic compounds studied. Immunostaining with an anticentromere antibody showed that >80% of MN induced gave a centromere-positive signal. Similarly, fluorescence in situ hybridization (FISH) analysis with alphoid centromere probes showed a positive hybridization signal, indicating that all compounds analyzed are aneugenic. Chemical modification of the N in the heterocyclic aromatic amine served us to suggest a relationship between the structure and the aneugenic effects of the compounds analyzed. Our findings indicate that benzoxazinoids could be potential genotoxins for human cells. PMID- 20006733 TI - The development of RAD51C, Cystatin A, p53 and Nrf2 luciferase-reporter assays in metabolically competent HepG2 cells for the assessment of mechanism-based genotoxicity and of oxidative stress in the early research phase of drug development. AB - Four different mechanism-based high-throughput luciferase-reporter assays were developed in human HepG2 cells, which contain phase I and II metabolic activity and a functionally active p53 protein. The promoter regions of RAD51C and Cystatin A, as well as the responsive element of the p53 protein, were selected for the generation of the genotoxicity reporter assays. Moreover, a luciferase based reporter assay was generated that measures the activation of the Nrf2 oxidative stress pathway. Validation with respect to the ECVAM compound list [D. Kirkland, P. Kasper, L. Muller, R. Corvi, G. Speit, Recommended lists of genotoxic and non-genotoxic chemicals for assessment of the performance of new or improved genotoxicity tests: a follow-up to an ECVAM workshop, Mutat. Res. 653 (2008) 99-108] resulted in an overall sensitivity of the HepG2 genotoxicity reporter assays for genotoxicity of 85% (17/20). The specificity and predictivity were high with 81% (34/42) and 82% (51/62), respectively. Various compounds had a positive score although metabolic activation was needed. The HepG2 reporter data were also compared with the available data on bacterial mutagenicity (Ames test), in vitro clastogenicity and in vivo clastogenicity for an additional set of 192 compounds. The predictivity for mutagenicity results was 74% (sensitivity, 61%, 30/49; specificity, 80%, 77/96) and for in vitro clastogenicity 59% (sensitivity, 45%, 35/78; specificity 83%, 38/46). The correlation between results from the HepG2 genotoxicity reporter assays and in vivo clastogenicity was much higher with 77% (sensitivity, 74%, 28/38; specificity 81%, 26/32). Results from the Nrf2 reporter assay showed that a large number of genotoxic compounds activated the Nrf2 oxidative stress pathway. In conclusion, four high-throughput mechanism based reporter assays in the HepG2 cell line were developed, which can be applied for screening in the early research phase of drug development. The use of these assays in combination with the previously validated Vitotox and RadarScreen assays will certainly reduce the attrition rate due to genotoxicity in the developmental phase of drug development. PMID- 20006734 TI - Ascorbic acid 2-glucocide reduces micronucleus induction in distant splenic T lymphocytes following head irradiation. AB - PURPOSE: Evidence from in vivo studies suggests there are enhanced radiation effects in abscopal regions after local head gamma ray irradiation. Splenocyte apoptosis and T lymphocyte micronuclei were induced at higher rates than what would be estimated given the dose at a shielded, distant position. In addition, we evaluated the radio-protective effects of ascorbic acid, acting as a radical scavenger on enhanced radiation effects in the shielded spleen following local head irradiation. METHODS AND MATERIALS: The heads of C3H mice were exposed to gamma-rays (10-20Gy), while the other parts of the body were shielded with a 5cm thick lead block. The effective dose for the spleen was calculated at 1.0-2.0Gy. Splenocytes were isolated 24h after cranial irradiation and their apoptosis was measured with an Elisa kit (Roche). The induction of T lymphocyte micronuclei was studied using the cytokinesis-block micronucleus assay. The ascorbic acid glucoside, 2-O-alpha-d-glucopyranosyl-l-ascorbic acid (AA-2G), was orally administered to mice 1h before whole body irradiation. The radio protective effects of AA-2G were estimated by comparing the induction of splenocyte damage (by apoptosis) and micronucleus induction. RESULTS: The splenocyte damage, as measured by the above two methods, was more excessive than what would be expected given exposure to 1.0-2.0Gy of radiation. Our results suggest that the effects were enhanced in a distant, non-irradiated organ after localized irradiation. Plasma ascorbic acid concentrations were increased 8-10x over control. Treatment with ascorbic acid slightly protected mouse splenocytes from the induction of apoptosis by the enhanced effects of radiation in the abscopal region. However, ascorbic acid significantly inhibited micronucleus induction in splenic T lymphocytes following local head irradiation. CONCLUSIONS: Our results suggest that ascorbic acid effectively scavenged radiation-induced radicals and protected against the enhanced effects of radiation in an abscopal region after local head gamma ray irradiation. PMID- 20006735 TI - GADD45a-GFP GreenScreen HC assay results for the ECVAM recommended lists of genotoxic and non-genotoxic chemicals for assessment of new genotoxicity tests. AB - A recent ECVAM workshop considered how to reduce falsely predictive positive results when undertaking in vitro genotoxicity testing, and thus to avoid unnecessary follow-up with tests involving animals. As it was anticipated that modified versions of existing assays as well as new assays might contribute to a solution, an expert panel was asked to identify a list of chemicals that could be used in the evaluation of such assays. Three categories of test chemicals were chosen comprising a total of 62 compounds. This paper provides test results for these chemicals using the GreenScreen HC assay. All tests were carried out in triplicate, by multiple operators, with and without S9, using invariant protocols. Group 1 chemicals should be detected as positive in in vitro mammalian cell genotoxicity tests: 18/20 (90%) were reproducibly positive in GreenScreen HC. Group 2 chemicals should give negative results in in vitro genotoxicity tests: 22/23 (96%) were reproducibly negative in GreenScreen HC. Overall concordance for Groups 1 and 2 is 93%. Group 3 chemicals should give negative results in in vitro mammalian cell genotoxicity tests, but have been reported to induce chromosomal aberrations or Tk mutations in mouse lymphoma cells, often at high concentrations or at high levels of cytotoxicity: 13/17 (76%) were reproducibly negative in GreenScreen HC. Of the four positive compounds in Group 3, p-nitrophenol was only positive at the top dose (10mM), 2,4-DCP is an in vivo genotoxin, and two chemicals are antioxidant compounds that may be acting as pro oxidants in the hyperoxic conditions of cell culture. Overall, these predictive figures are similar to those from other studies with the GreenScreen HC assay and confirm its high specificity, which in turn minimizes the generation of falsely predictive positive results. PMID- 20006736 TI - Dealing with DNA damage: relationships between checkpoint and repair pathways. AB - Cell cycle checkpoint activation and DNA repair pathways govern genomic stability after genotoxic stress. Genotoxic insult results in activation of an interwoven network of DNA damage checkpoints and DNA repair pathways. Post-translational modifications on a number of proteins involved in both checkpoint activation and DNA repair play an important role in this cellular response. Genotoxic stress can induce a wide variety of DNA lesions. Among these DNA alterations are double stranded breaks and single-stranded DNA gaps. Repair of these DNA alterations requires damage recognition and resection. Here we discuss how DNA repair and DNA damage checkpoints cooperate and deal with DNA damage. Processing of DNA lesions by structure-specific nucleases results in DNA-protein intermediates, which form the basis for checkpoint activation and DNA repair. Post-translational modifications like phosphorylation and ubiquitination modulate the DNA damage response in a spatial and temporal manner. Cell cycle-dependent regulation additionally plays a key role in the regulation of both DNA repair and checkpoint activation. We highlight recent advances in in vivo imaging that greatly expand our knowledge on the relationships between DNA damage checkpoints and DNA repair. PMID- 20006737 TI - Pigment epithelium-derived factor inhibits vascular endothelial growth factor-and interleukin-1beta-induced vascular permeability and angiogenesis in retinal endothelial cells. AB - Increased vascular permeability associated with retinal vascular leakage is known to occur in patients with diabetes, and contributes to endothelial barrier dysfunction. The purpose of this study was to examine the effect of pigment epithelium-derived factor (PEDF) on signaling cascade in porcine retinal endothelial cells (PREC) related to permeability and angiogenesis induced by vascular endothelial growth factor (VEGF)-and interleukin-1beta (IL-1beta). PREC were exposed to VEGF, IL-1beta and PEDF at different concentrations, and in vitro permeability was assessed by solute flux assay using 70-kDa RITC-dextran. Angiogenic assays such as proliferation, migration and tube formation were determined by MTT, wound-scratch method and on-gel assay system respectively. To explore the signaling pathways behind VEGF-and IL-1beta-induced PREC permeability, an inhibitor assay was carried out using PP2, a Src kinase inhibitor. Further, Src activity was assessed by transient transfection assay using constitutively active (CA) and dominant negative (DN) Src mutants. We report that VEGF-and IL-1beta-stimulates permeability, in a dose and time dependent manner and PEDF inhibits the VEGF-and IL-1beta-induced PREC permeability. In addition, PEDF inhibits the VEGF-and IL-1beta-induced endothelial cell proliferation, migration and tube formation. In addition, overexpression of DN Src blocked both VEGF-and IL-1beta-stimulation of permeability, proliferation and migration, while overexpression of CA Src overpowers the inhibitory action of PEDF on permeability, proliferation and migration. These results demonstrate that PEDF may inhibit the VEGF-and IL-1beta induced permeability and angiogenesis via Src-dependent pathway. PMID- 20006738 TI - Somatic mutations of mitochondrial genome in hepatocellular carcinoma. AB - Somatic mutations have been identified in mitochondrial DNA (mtDNA) of various human primary cancers. However, their roles in the pathophysiology of cancers are still unclear. In our previous study, high frequency of somatic mutations was found in the D-loop region of mtDNA of hepatocellular carcinomas (HCCs). In the present study, we examined 44 HCCs and corresponding non-cancerous liver tissues, and identified 13 somatic mutations in the coding region of mtDNAs from 11 HCC samples (11/44, 25%). Among the 13 mtDNA mutations, six mutations (T6787C, G7976A, A9263G, G9267A, A9545G and A11708G) were homoplasmic while seven mutations (956delC, T1659C, G3842A, G5650A, 11032delA, 12418insA and a 66bp deletion) were heteroplasmic. Moreover, the G3842A transition created a premature stop codon and the 66bp deletion could omit 22 amino acid residues in the NADH dehydrogenase (ND) subunit 1 (ND1) gene. The 11032delA and 12418insA could result in frame-shift mutation in the ND4 and ND5 genes, respectively. The T1659C transition in tRNA(Val) gene and G5650A in tRNA(Ala) gene were reported to be clinically associated with some mitochondrial disorders. In addition, the T6787C (cytochrome c oxidase subunit I, COI), G7976A (COII), G9267A (COIII) and A11708G (ND4) mutations could result in amino acid substitutions in the highly conserved regions of the affected mitochondrial genes. These mtDNA mutations (10/13, 76.9%) have the potential to cause mitochondrial dysfunction in HCCs. Taken these results together, we suggest that there may be a higher frequency of mtDNA mutations in HCC than in normal liver tissues from the same individuals. PMID- 20006739 TI - Contribution of the FAD and quinone binding sites to the production of reactive oxygen species from Ascaris suum mitochondrial complex II. AB - Reactive oxygen species (ROS) production from mitochondrial complex II (succinate quinone reductase, SQR) has become a focus of research recently since it is implicated in carcinogenesis. To date, the FAD site is proposed as the ROS producing site in complex II, based on studies done on Escherichia coli, whereas the quinone binding site is proposed as the site of ROS production based on studies in Saccharomyces cerevisiae. Using the submitochondrial particles from the adult worms and L(3) larvae of the parasitic nematode Ascaris suum, we found that ROS are produced from more than one site in the mitochondrial complex II. Moreover, the succinate-dependent ROS production from the complex II of the A. suum adult worm was significantly higher than that from the complex II of the L(3) larvae. Considering the conservation of amino acids crucial for the SQR activity and the high levels of ROS production from the mitochondrial complex II of the A. suum adult worm together with the absence of complexes III and IV activities in its respiratory chain, it is a good model to examine the reactive oxygen species production from the mitochondrial complex II. PMID- 20006740 TI - Mitochondrial DNA haplogroup 'R' is associated with Noonan syndrome of south India. AB - Mutations in PTPN11 gene was responsible for approximately 50% of the Noonan syndrome (NS), however, we did not find any mutation in PTPN11 in any of seven NS patients analysed. Whereas, the complete mtDNA sequencing revealed 146 mutations, of which five, including one heteroplasmic (A11144R; Thr-->Ala) non-synonymous mutation, were novel and exclusively observed in NS patients. Interestingly all the seven probands and their maternal relatives were clustered under a major haplogroup R and its novel sub-haplogroups (R7b1b, R30a1, R30c, T2b7, U9a1) exclusive in NS, therefore we strongly suggest that these haplogroups may influence NS in South Indian populations. PMID- 20006741 TI - Thermal aggregation of glycated bovine serum albumin. AB - Aggregation and glycation processes in proteins have a particular interest in medicine fields and in food technology. Serum albumins are model proteins which are able to self-assembly in aggregates and also sensitive to a non-enzymatic glycation in cases of diabetes. In this work, we firstly reported a study on the glycation and oxidation effects on the structure of bovine serum albumin (BSA). The experimental approach is based on the study of conformational changes of BSA at secondary and tertiary structures by FTIR absorption and fluorescence spectroscopy, respectively. Secondly, we analysed the thermal aggregation process on BSA glycated with different glucose concentrations. Additional information on the aggregation kinetics are obtained by light scattering measurements. The results show that glycation process affects the native structure of BSA. Then, the partial unfolding of the tertiary structure which accompanies the aggregation process is similar both in native and glycated BSA. In particular, the formation of aggregates is progressively inhibited with growing concentration of glucose incubated with BSA. These results bring new insights on how aggregation process is affected by modification of BSA induced by glycation. PMID- 20006742 TI - Blast injuries of large tyres: case series. AB - BACKGROUND: Severe blast injuries of large tyres are similar to those resulting from landmine explosions with neither thermal nor chemical effects. Little has been written about the destructive nature of these blasts. AIMS: To evaluate our clinical management of patients involved in large tyre blasts. PATIENTS AND METHODS: All patients who had tyre blast injuries and were admitted to Al-Ain or Tawam Hospitals between March 2003 and September 2009 were retrospectively studied. Clinical presentation, mechanism of injury, management, and outcome were reviewed. RESULTS: Seven male patients were studied. They had a median (range) age of 38 (20-53) years. Four patients (57%) were inflating the tyre when it suddenly exploded. On arrival to the hospital, two patients were unconscious with GCS of 3/15. Six patients (86%) had head/face trauma. Three patients had multiple injuries to different body parts (43%). The median (range) injury severity score was 14 (10-33). Four patients (57%) were operated on. Five patients were admitted to the ICU with a median (range) ICU stay of 2 (1-2) days. The median (range) total hospital stay was 3 (1-14) days. Two patients died (overall mortality 29%). CONCLUSIONS: The high energy produced by large tyre blasts may cause severe injuries leading to high morbidity and mortality. Preventive occupational methods should be adopted and implemented at the work place. PMID- 20006743 TI - Functional analysis of a type 1 parathyroid hormone receptor intracellular tail mutant [KRK(484-6)AAA]: effects on second messenger generation and cellular targeting. AB - The parathyroid hormone receptor type 1 (PTHR1) is activated by parathyroid hormone (PTH) and PTH-related protein (PTHrP) and primarily signals via intracellular pathways involving adenylyl cyclase and phospholipase C. The intracellular tail domain of the PTHR1 contributes to G protein subunit coupling that is important for second messenger signalling. In addition, the intracellular domain has a potential nuclear localization sequence (NLS) that, if functional, could point to an intracrine role for the receptor. In the present study, we have utilized 2 sets of constructs that employ either a [KRK(484-486)AAA](3Ala) mutation in the putative NLS or the non-mutant counterpart and included (a) the full-length rat PTHR1 with FLAG and c-myc epitope tags at the N-terminus and C terminus, respectively (designated as PTHR1(3Ala)-TAG and PTHR1-TAG); and (b) only the putative NLS-containing intracellular domain (471-488), with green fluorescent protein (GFP) fused to the C-terminus (designated as GFP-(3Ala)471 488 or GFP-471-488). Porcine kidney LLC-PK1 cells stably expressing the PTHR1(3Ala)-TAG exhibited reduced signalling via both cAMP and cytosolic calcium transients in spite of greater cell surface expression relative to cells expressing PTHR1-TAG. We also examined the ability of the intracellular tail to influence the cellular localization of a heterologous protein. LLC-PK1 cells transiently transfected with GFP-471-488, exhibited increased fluorescence within the nucleus, relative to cells transfected with GFP alone that was not observed when cells were transiently transfected with the mutated construct, GFP-(3Ala)471 488. However, LLC-PK1 cells transiently transfected with either the full-length PTHR1-TAG or the PTHR1(3Ala)-TAG constructs did not exhibit nuclear localization of these receptors. Moreover, mouse osteoblast-like cells (MC3T3-E1) transiently expressing PTHR1-TAG also failed to demonstrate nuclear localization, although both full-length PTHR1 constructs exhibited plasma membrane immunofluorescence in both cell lines. Thus, the 484-486 sequence is critical for the full signalling responsiveness of the intact PTHR1, but the putative nuclear localization signal may not function as such within the intact receptor. PMID- 20006744 TI - Focal neuromodulation for the treatment of depression. PMID- 20006750 TI - Community-associated MRSA finally gets to central New York. PMID- 20006745 TI - Pioneering first steps and cautious conclusions. PMID- 20006755 TI - A perspective on 360-degree evaluations. PMID- 20006756 TI - Chorioamnionitis, surfactant, and lung disease in very low birth weight infants. PMID- 20006757 TI - Rectal perceptual hypersensitivity: a biomarker for pediatric irritable bowel syndrome. PMID- 20006758 TI - Decisions in the gray zone: evidence-based or culture-based? PMID- 20006764 TI - An ethical and policy analysis of elective transplantation for metabolic conditions diagnosed by newborn screening. PMID- 20006765 TI - Low body adiposity and high leptinemia in breast-fed infants born small-for gestational-age. AB - Breast-fed infants born small-for-gestational-age (SGA) (n = 28) were found to develop lower adiposity across early infancy, when compared with appropriate-for gestational age infants (n = 46). The SGA infants were also found to have 2-fold higher (P < .001) levels of circulating leptin at 4 months. This paradoxical hyperleptinemia in early infancy may contribute to a program that produces a process from fetal underweight to adult overweight. PMID- 20006766 TI - Massive pulmonary embolism in children. AB - We present 3 children with massive pulmonary embolism and review 17 recent pediatric reports. Malignancies were a frequent cause (40%), and sudden death was common (60%). Compared with adults, diagnosis was more likely to be made at autopsy (P < .0001), more children were treated with embolectomy/thrombectomy (P = .0006), and mortality was greater (P = .03). PMID- 20006767 TI - Incidence of invasive community-onset Staphylococcus aureus infections in children in Central New York. AB - We determined the incidence of invasive community-onset Staphylococcus aureus infections, clinical characteristics, and antibiotic susceptibilities in 128 hospitalized children in central New York. The prevalence of invasive S aureus infections in our institution remained <1% between 1996 and 2006, although the proportion of methicillin-resistant S aureus infections significantly increased. PMID- 20006770 TI - Autoimmune focal encephalitis shows marked hypermetabolism on positron emission tomography. AB - A 22-month-old toddler presented with involuntary movements, hemiparesis, and behavioral changes. Magnetic resonance imaging showed no abnormality, but positron emission tomography (PET) showed focal hypermetabolism. By immunohistochemical technique with the patient's sera in control brain sections, autoantibodies recognizing the same areas as found by PET scanning were identified and disappeared after intravenous immunoglobulin therapy. PET scanning may be useful in the diagnosis of autoimmune encephalitis. PMID- 20006771 TI - Pulmonary hypoplasia in a newborn with Goldenhar syndrome. PMID- 20006769 TI - Brown recluse spider (Loxosceles reclusa) envenomation leading to acute hemolytic anemia in six adolescents. AB - Loxosceles reclusa (brown recluse spider) bites often cause local envenomation reactions; however, acute hemolysis from systemic loxoscelism is rare. To highlight this important diagnostic consideration for unexplained hemolysis in areas endemic for brown recluse spiders, we report on 6 adolescents with acute hemolytic anemia from presumed L reclusa bites. PMID- 20006772 TI - Perimenarchal menorrhagia: evaluation and management. PMID- 20006773 TI - Amoxicillin treatment increases rate of late recurrence of acute otitis media in young children. PMID- 20006774 TI - Ibuprofen elevates total bilirubin in preterm infants. PMID- 20006775 TI - Xylitol syrup can reduce dental caries progression in young children. PMID- 20006776 TI - Fever and hypoxia predict radiographic pneumonia among children with wheezing. PMID- 20006777 TI - Ambulatory treatment of infants with presumed febrile urinary tract infection may be feasible. PMID- 20006778 TI - Amoxicillin/potassium clavulanate is effective treatment for acute bacterial sinusitis in children. PMID- 20006780 TI - Violence in media research. PMID- 20006779 TI - Successful prevention of severe infection in Japanese siblings with interleukin-1 receptor-associated kinase 4 deficiency. PMID- 20006782 TI - Orotracheal tube length in ELBW neonates. PMID- 20006784 TI - Cilia. Preface. PMID- 20006785 TI - Aging and the kidney: introduction. PMID- 20006786 TI - The aging pandemic: demographic changes in the general and end-stage renal disease populations. AB - The aging of the US population, and Western society populations in general, has been of concern to demographers and health planners for a number of years. The implications of this demographic shift include both increased economic and health care burdens for the United States. The growth of the end-stage renal disease population in the 1980s and 1990s suggested massive increases in this very expensive and frail population. However, recent incidence rates suggest that future growth may not be as great as thought just a few years ago. PMID- 20006787 TI - Chronic inflammation potentiates kidney aging. AB - Chronic inflammation, characterized by increased serum levels of tumor necrosis factor-alpha, interleukin-6, C-reactive protein, and plasminogen activator inhibitor-1, and the presence of inflammatory-related diseases, are seen commonly in aging. Both the dysregulation of immune cells and phenotypic changes in parenchymal cells may contribute to chronic inflammation in aging. Moreover, senescent cells are an important source of inflammatory factors. Oxidative stress, via activation of p38 and c-Jun N-terminal kinase and induction of cell senescence, is likely to play a critical role in inflammation. Endoplasmic reticulum stress also may be present in aging and be involved in inflammation. Advanced glycation end products also are important contributors to inflammation in aging. Because the kidney is a major site for the excretion, and perhaps the degradation, of advanced glycation end products and small inflammatory molecules, reduced renal function in aging may promote oxidative stress and inflammation. Chronic inflammation in turn may potentiate the initiation and progression of lesions in the aging kidney. PMID- 20006788 TI - Sexual dimorphism, the aging kidney, and involvement of nitric oxide deficiency. AB - Females develop less age-dependent loss of renal function, in part because of cardiorenal protective effects of estrogens. The low androgen level in women also may be protective, although the animal and clinical data are controversial. Both estrogen and androgens act through multiple mechanisms, sometimes beneficial, sometimes damaging, which makes it difficult to predict the impact of hormone replacement therapy in an aging population. Nitric oxide (NO) deficiency contributes to age-dependent cardiovascular risk and kidney damage in animal models. The increased oxidative stress of aging impacts at multiple sites in the NO biosynthetic pathway to decrease NO production/action. Endothelial dysfunction develops with aging and is delayed in women, in association with a delayed increase in asymmetric dimethylarginine. Animal data suggest that the aging kidney develops NO deficiency because of changes in the neuronal NO synthase. Relative preservation of NO production in females contributes to the better cardiovascular and renal responses to aging. PMID- 20006789 TI - Urinary concentration and dilution in the aging kidney. AB - Aged people and rats have a reduced ability to maximally concentrate their urine. Many of the key transport proteins that contribute to urine concentrating ability are reduced in the medulla of aged rats. The reductions in the abundances of water, sodium, and urea transport proteins, and their reduced response to water restriction, contributes to the reduced ability of aged rats to concentrate their urine and conserve water. If similar mechanisms occur in human kidneys, it would provide a molecular explanation for the reduced concentrating ability in aging and may provide opportunities for novel therapeutic approaches to improve urine concentrating ability. PMID- 20006790 TI - Podocytes and glomerular function with aging. AB - Kidney function declines with age in association with the development of age associated glomerulosclerosis. The well-established structural and functional changes with age are reviewed briefly. The modification of aging pathology by calorie restriction is discussed. The role of the podocyte as a critical cell in the aging process is considered, using animal models and human biopsy material. Newer data on changes in gene expression and possible changes in biology in the glomerulus are discussed. There is speculation on the implications of this change in biology for human disease and progression. PMID- 20006792 TI - Hypertension and its management in the elderly. AB - Many trials focused on cardiovascular outcomes demonstrate that reduction in blood pressure to levels below 140/90 mm Hg reduce cardiovascular events including stroke and myocardial infarction. There are very few such trials, however, in the elderly cohort, especially among those aged 70 and older. In the few outcome trials that have been completed in this older age group, systolic blood pressures in the range between 140 and 149 mm Hg demonstrate a clear reduction in cardiovascular events. Moreover, among the subgroup that has a vasculature that allows a systolic pressure to go below 140 mm Hg without cognitive side effects, ie, somnolence, memory loss, etc, does even better than those whose blood pressures are in the 140 to 149 mm Hg range. Thus, titration of systolic blood pressure goals in the elderly should strive for a goal of less than 140 mm Hg, and if not achievable without side effects, compromise to below 150 mm Hg. PMID- 20006791 TI - Identifying advanced glycation end products as a major source of oxidants in aging: implications for the management and/or prevention of reduced renal function in elderly persons. AB - Aging is characterized by increasing inflammation and oxidant stress (OS). Reduced renal function was present in more than 20% of normal-aged individuals sampled in the National Health and Nutrition Examination Survey (NHANES) cross sectional study of the US population. Longitudinal studies in the United States and Italy showed that renal function does not decline in some individuals, suggesting that a search for causes of the loss of renal function in some persons might be indicated and interventions to reduce this outcome should be sought. Because advanced glycation end products (AGEs) induce both inflammation and OS, accumulate with age, and primarily are excreted by the kidney, one outcome of reduced renal function in aging could be decreased AGE disposal. The build-up of AGEs with reduced renal function could contribute to inflammation, increased oxidant stress, and accumulation of AGEs in aging. In fact, results from a longitudinal study of normal aging adults in Italy showed that the most significant correlation with mortality was the level of renal function. A clear link between inflammation, OS, AGEs, and chronic disease was shown in studies of mice that showed that reduction of AGE levels by drugs or decreased intake of AGEs reduces chronic kidney disease (CKD) and cardiovascular disease of aging. The data support a role for AGEs in the development of renal lesions in aging mice and reveal that AGEs in the diet are very important contributors to renal and cardiovascular lesions. AGEs signal through two receptors, one of which is anti-inflammatory (AGER1) and the other is proinflammatory (RAGE). Overexpression of AGER1 protects against OS and acute vascular injury. The reduction of AGEs in the diet is as efficient in preventing aging-related cardiovascular and renal lesions in mice as that seen with calorie restriction. Studies in normal adults of all ages and those with CKD suggest that the findings in mice may be directly applicable to both aging and CKD. Namely, the dietary content of AGEs determines the serum levels of AGEs and inflammatory mediators and urine AGE levels in both normal subjects and CKD patients. Importantly, reduction of AGEs controls these changes in both normal subjects and CKD patients, and the phenotypic changes in AGER1 are reduced in CKD patients by decreasing the amount of AGEs consumed with the diet. These data suggest that the changes in renal function in normal aging may be subject to control and this subject deserves renewed attention. PMID- 20006795 TI - Renal osteodystrophy and aging. AB - The bone disease seen in our aging dialysis population is a complex mixture of osteoporosis and renal osteodystrophy. Attention must be paid to both of these issues. Hip fractures are increased with aging and this increase is further aggravated by renal failure. Preventive management with Vitamin D and bisphosphonates is reviewed. PMID- 20006796 TI - Management of elderly patients with end-stage kidney disease. AB - The number of elderly patients requiring renal replacement therapy will continue to increase in the future. Previously, these patients were believed to have a poor prognosis on renal replacement therapy and therefore they were treated with conservative measures. Although there are no controlled randomized studies, an increasing amount of literature indicates that elderly persons with end-stage kidney disease are a heterogeneous population and that, in many of them, dialysis provides a good quality of life and survival. Both hemodialysis and peritoneal dialysis have their own advantages and disadvantages and therefore it is important to individualize treatment to achieve the best result for each patient, taking into account the patient's wishes, local circumstances, availability of dialysis centers, and proper social support. Home treatments are the most suitable for frail elderly patients. In the latter population, peritoneal dialysis may be the method of choice because it can be performed by visiting nurses, thus allowing the patient to receive treatment at home without increasing the overall cost. PMID- 20006793 TI - Disorders of lipid metabolism and chronic kidney disease in the elderly. AB - The growing population of elderly with chronic kidney disease (CKD) is at greater risk for cardiovascular disease given an independent risk of CKD, as well as from added dyslipidemia of aging and renal dysfunction. Changes in lipid metabolism with more isodense and high-dense, triglyceride-rich particles, low high-density lipoprotein cholesterol, and increased triglyceride levels occur with CKD and aging, which are noted to have significant atherogenic potential. In addition, lipid abnormalities may lead to the progression of CKD. Cardiovascular mortality in the end-stage renal disease population is more than 10 times higher than the general population. Treatment of dyslipidemia in the general population suggests important benefits both in reducing cardiovascular risk and in the prevention of cardiovascular disease. Secondary analyses of elderly subgroups of various large prospective studies with statins suggest treatment benefit with statin use in the elderly. Similarly limited data from secondary analyses of CKD subgroups of larger prospective trials using statins also suggest a possible benefit in cardiovascular outcomes and the progression of kidney disease. However, randomized trials have yet to confirm similar benefits and targets of treatment for dyslipidemia in the elderly with CKD and end-stage renal disease. Treatment in the elderly with CKD should be individualized and outweigh risks of side effects and drug-drug interactions. There is a need for further specific investigation of dyslipidemia of CKD in the aging population in relation to renal disease progression and cardiovascular outcome. PMID- 20006794 TI - Kidney transplantation in the elderly. AB - There is an increase in the older incident end-stage renal disease population that is associated with an increasing prevalence of end-stage renal disease in the United States. This trend is paralleled by an increasing rate of kidney transplantation in the elderly. Although patient survival is lower in older versus younger kidney recipients, the elderly benefit from a reduction in mortality rate and improved quality of life with transplantation compared with dialysis. Immunologic, physiologic, and psychosocial factors influence transplant outcomes and should be recognized in the care of the elderly transplant patient. In this review, we discuss transplantation in the elderly patient, particularly the topics of access to transplantation, patient and graft survival, the impact of donor quality on transplant outcomes, immunology and immunosuppression of aging, and ethical considerations in the development of an equitable organ allocation scheme. PMID- 20006797 TI - Longitudinal measurements of renal function. AB - Cross-sectional studies have long suggested that renal function declines with age among adults. However, there are limited longitudinal studies with data on within individual changes in renal function over time. The Baltimore Longitudinal Study of Aging provided some of the first data on within-person changes in renal function over time. In a subsample of this cohort without renal disease, followed up to 23 years, creatinine clearance declined by a mean of -0.75 mL/min/1.73 m(2) per year. Interestingly, 35% of individuals in this study did not experience a decline in renal function. Similar findings from longitudinal studies of patients with established chronic kidney disease have been reported. Among 1,094 participants with up to 10 years of follow-up evaluation in the African-American Study of Kidney Disease, the mean annual rates of decline in estimated glomerular filtration rate were -2.11 mL/min/1.73 m(2) and -1.50 mL/min/1.73 m(2) during the trial and observational phases, respectively. Of participants with at least 7 years of follow-up evaluation, 33.5% experienced a mean annual decline in estimated glomerular filtration rate of less than -1 mL/min/1.73 m(2). More studies are needed to better define longitudinal changes in renal function. Such data will provide important information for guiding treatment choices for patients, research endeavors, and health policy. PMID- 20006798 TI - Acute kidney injury in the elderly: problems and prospects. AB - Acute kidney injury (AKI) usually is defined as a decline in glomerular filtration rate over hours to days that can occur either in a person with previously normal kidney function, or in the setting of pre-existing chronic kidney disease. The incidence of AKI has increased steadily in recent years, and this increase is associated strongly with advancing age in the population because epidemiologic data indicate that AKI is much more common in the elderly. In the aging population there is heightened susceptibility to drug toxicity, partially owing to altered drug pharmacokinetics and pharmacodynamics. Furthermore, the elderly consume twice as many medications overall, including nephrotoxic agents, compared with younger patients. The vasodilatory increase in the renal plasma flow and glomerular filtration rate that constitutes renal functional reserve is reduced markedly in healthy elderly individuals, compromising renal adaptation after acute ischemia, and heightening susceptibility to AKI. Age-related alterations in renal tubular function also may heighten susceptibility to AKI because renal sodium conservation in response to dietary sodium restriction is attenuated in the elderly. A recently published study showed that elderly subjects with AKI, particularly those with chronic kidney disease, are more likely to develop end-stage renal disease. The increasing prevalence and extended clinical consequences of AKI in the elderly are of great concern from a clinical and public health perspective. PMID- 20006799 TI - Surgical revascularization. Introduction. PMID- 20006800 TI - Impact of comorbidities on decision-making in chronic critical limb ischemia. AB - Given both the high-risk nature of the critical limb ischemia (CLI) patient population, as well as the variety of available treatment options, the ability to use preprocedure variables to predict a given outcome has become increasingly important. The three main advantages associated with accurate risk stratification include: (1) improved clinical decision-making based on realistic patient and physician expectations, (2) ability to benchmark individual practitioners and institutions by comparing their outcomes with expected outcomes specific to the actual patients being treated, and (3) creation of objective performance goals for the evaluation of new treatment modalities in specific patient populations. Patients with CLI have multiple comorbidities that can impact greatly on the outcomes of attempted revascularization. The PREVENT III CLI Risk Score was developed as a tool to estimate amputation-free survival at 1 year in patients undergoing vein bypass surgery for limb salvage, and has now been validated using data from >3,000 cases. The score incorporates five baseline clinical variables dialysis, tissue loss, age, hematocrit, and coronary artery disease-and assigns patients into three distinct risk groups. Patients in the high-risk subgroup (8.2% of total number of patients evaluated) experienced <50% amputation-free survival at 1 year. This tool may be valuable in assisting both the surgeon and patient faced with treatment decisions in CLI. PMID- 20006801 TI - Risk factors for lower-extremity vein graft failure. AB - Lower-extremity vein graft failure causes significant morbidity, increases health care costs, and negatively impacts patient quality of life. Identification of risk factors is essential for patient selection, risk factor modification, and identifying individuals who would benefit from more stringent surveillance protocols. Risk factors can be considered as either patient-related or technical. Here we discuss the patient-related risk factors for vein graft failure. Nontechnical factors related to the indication for operation include operation after a previously failed graft, or redo bypass, critical limb ischemia, and infection. Risk factors for vein graft failure are distinct from the risk factors for cardiovascular events. Young age and African American and Hispanic race are risk factors for lower-extremity vein graft failure. Hypercoaguable and inflammatory states also increase risk for vein graft failure. Therapy with statins is indicated in patients with peripheral atherosclerosis and may have beneficial effects on vein graft function, although further studies are needed in this area. PMID- 20006802 TI - Technical factors in lower-extremity vein bypass surgery: how can we improve outcomes? AB - Lower-extremity vein bypass surgery has been a standard operation in the armamentarium of vascular surgeons for more than 4 decades. Yet there remains surprising heterogeneity in the utilization, techniques, and outcomes associated with this procedure in current practice. Despite improvements in surgical technique and careful postoperative surveillance, vein graft failure remains a significant clinical problem affecting up to 50% of patients within 5 years. Experience, clinical judgment, creativity, and technical precision are required to optimize long-term results. Many factors, including patient-specific comorbidities, and variable biologic responses in the venous conduit, influence the ultimate outcome. Technical factors, however, play a dominant role in determining clinical success. Multiple single-center reports, and more recent multicenter trials, have clearly demonstrated that conduit selection and quality is the most critical element. An adequate caliber (>/=3.5 mm), good quality great saphenous vein is the optimal graft for infrainguinal bypass. Contralateral great saphenous vein, arm vein, and lesser saphenous vein are the next most desirable conduits. Graft configuration (reversed, nonreversed, or in situ) appears to have little influence on outcomes per se. Shorter grafts have improved patency. Inflow can be improved by surgical or endovascular means if necessary, and distal-origin grafts perform as well as those originating from the common femoral artery. The selected outflow vessel should supply unimpeded runoff to the foot, conserve conduit length, and allow for adequate soft-tissue coverage of the graft and a simplified surgical exposure. This review summarizes the data linking technical factors with graft patency, highlighting the importance of surgical judgment and operative planning in the current practice of infrainguinal bypass surgery. PMID- 20006803 TI - Secondary bypass after infrainguinal bypass graft failure. AB - Patients who suffer failure of a previous infrainguinal bypass graft often present with recurrent ischemia requiring secondary revascularization for limb salvage. These patients pose major challenges for the vascular surgeon. Management strategies vary with the time interval from bypass, the functional status of the patient, the degree of ischemia and the availability of autogenous vein. This article reviews the treatment options for these difficult patients and the results which can be achieved. PMID- 20006804 TI - Hybrid procedures and distal origin grafts. AB - Endovascular techniques for treatment of infrainguinal arterial occlusive disease continue to dominate care for most symptomatic vascular patients. Diabetic patients with gangrene represent a subset of patients who can benefit from a combination of open and percutaneous techniques because of the multilevel nature of the disease. Most commonly, these involve surgical treatment of the infrageniculate disease with a distal origin bypass coupled with endovascular treatment of the ipsilateral superficial femoral artery. These approaches allow care to be delivered in a single setting, avoiding issues of staged procedures. Although published results from hybrid treatment are limited, the obvious patient benefit ensures that these techniques will continue to be important for vascular patient care. PMID- 20006805 TI - What is the optimum perioperative drug therapy following lower-extremity vein bypass surgery? AB - While endoluminal procedures are now commonly done for symptomatic peripheral arterial disease, vein bypass remains the gold standard for revascularization. Lower extremity vein bypass procedure success is dependent on patient factors, surgical judgment and technique, including use of medications. Cardioprotective medications have proven efficacy to decrease morbidity and mortality, but their use to improve graft patency is less well known. We review the up to date use of medications with known vascular effects that may promote graft success, as well as decrease cardiovascular events in this high risk patient group. PMID- 20006806 TI - Detection of imminent vein graft occlusion: what is the optimal surveillance program? AB - The prediction of infrainguinal vein bypass failure remains an inexact judgment. Patient demographics, technical factors, and vascular laboratory graft surveillance testing are helpful in identifying a high-risk graft cohort. The optimal surveillance program to detect the bypass at risk for imminent occlusion continues to be developed, but required elements are known and include clinical assessment for new or changes in limb ischemia symptoms, measurement of ankle and/or toe systolic pressure, and duplex ultrasound imaging of the bypass graft. Duplex ultrasound assessment of bypass hemodynamics may be the most accurate method to detect imminent vein graft occlusion. The finding of low graft flow during intraoperative assessment or at a scheduled surveillance study predicts failure; and if associated with an occlusive lesion, a graft revision can prolong patency. The most common abnormality producing graft failure is conduit stenosis caused by myointimal hyperplasia; and the majority can be repaired by an endovascular intervention. Frequency of testing to detect the failing bypass should be individualized to the patient, the type of arterial bypass, and prior duplex ultrasound scan findings. The focus of surveillance is on identification of the low-flow arterial bypass and timely repair of detected critical stenosis defined by duplex velocity spectra criteria of a peak systolic velocity 300 cm/s and peak systolic velocity ratio across the stenosis >3.5-correlating with >70% diameter-reducing stenosis. When conducted appropriately, a graft surveillance program should result in an unexpected graft failure rate of <3% per year. PMID- 20006807 TI - Revision of vein bypass grafts: factors affecting durability of interventions. AB - Once the failing vein graft is identified and characterized, the clinician must choose the appropriate intervention to maintain graft patency. Limited by the single-institution, retrospective studies that are pervasive in this area, definitive data to guide these decisions are limited. In general, open surgical revisions appear to offer a modest benefit in primary patency, but likely at the cost of increased periprocedural morbidity. Although endovascular revisions are more prone to failure, these recurrent lesions are often amenable to reintervention so that the secondary patency rates for both endovascular and open interventions may be similar. Given this, endovascular intervention as an initial treatment modality seems a reasonable approach for favorable lesions. Factors associated with poor outcome for endovascular revision include longer lesions (stenosis >2 cm in length), multiple stenoses, lesions occurring within 3 months of graft placement, or interventions for graft thrombosis, where endovascular failures are high and open surgery as an initial approach is warranted. The optimum method for percutaneous intervention remains a shifting landscape. No techniques as of yet appear clearly superior to standard balloon angioplasty, but initial investigations would suggest that cutting balloons offer a modest improvement and are worthy of consideration. PMID- 20006808 TI - Bypass versus angioplasty in severe ischaemia of the leg (BASIL) trial: what are its implications? AB - Lack of Level I evidence from randomized controlled trials (RCT) means that the relative merits of surgical and endovascular revascularization strategies for severe limb ischemia (SLI) due to infrainguinal disease remain unclear. The Bypass versus Angioplasty in Severe Ischaemia of the Leg (BASIL) trial remains the only multicenter RCT to have compared the clinical and cost-effectiveness of bypass surgery (BSX)-first and balloon angioplasty (BAP)-first revascularization strategies for infrainguinal SLI. An intention to treat analysis shows that out to 2 years both strategies were associated with similar amputation-free (AFS) and overall survival (OS) rates, as well as improvements in health-related quality of life. In the short-term, BSX was significantly more morbid and expensive. However, for those patients who survived for 2 years after randomization, initial randomization to a BSX-first strategy was associated with a significant increase in subsequent OS of about 7 months and a nonsignificant increase in subsequent AFS of about 6 months. Vein BSX performed significantly better than prosthetic BSX in terms of AFS but not OS. For most patients BAP also appears preferable to prosthetic BSX. Patients who underwent BSX after a failed BAP-first strategy did not fare as well as those who received BSX as their first procedure. Patients who are expected to live less than 2 years should usually be offered BAP first, especially when the alternative is prosthetic BSX. Those expected to survive beyond this time horizon (approximately 75% of the BASIL cohort) should usually be offered BSX first, especially where vein is available. Further RCTs to confirm or refute these findings and recommendations are required. PMID- 20006809 TI - Does lower-extremity bypass improve quality of life? Is it cost effective? AB - Limb salvage, operative mortality and short and long term graft patency have long been important variables for assessment of infrainguinal arterial reconstructions. More recently it is now appreciated that patient and society oriented measures of outcomes in determining the effectiveness of any surgical procedure are also important. In particular, the impact of an intervention on a patient's overall quality of life (QOL) and whether it is cost effective in achieving improvement of QOL are questions of increasing importance from the patient's perspective and from the perspective of those who must make difficult health policy decisions. This chapter reviews the cost effectiveness and impact of infrainguinal bypass on patient perceived quality of life in patients with intermittent claudication and patients with critical limb ischemia. The data are not robust but suggest favorable impact of infrainguinal bypass in terms of both QOL and cost effectiveness in both patients with intermittent claudication and critical limb ischemia. PMID- 20006810 TI - World of shoulder and elbow surgery. PMID- 20006811 TI - Transscleral diode laser retinal photocoagulation for the treatment of threshold retinopathy of prematurity. AB - PURPOSE: To evaluate the outcomes of transscleral diode laser photocoagulation for treatment of threshold retinopathy of prematurity (ROP). METHODS: Retrospective interventional case series of infants with ROP treated at our center with transscleral diode laser retinal photocoagulation. All patients were followed for 2 months, and regression of ROP, unfavorable outcomes, and incidence of adverse effects were assessed. RESULTS: A total of 103 eyes of 52 infants with ROP at the threshold stage were treated with transscleral diode laser photocoagulation under topical anesthesia, without conjunctival incision. Supplemental transpupillary diode laser photocoagulation was used for posterior zone 2 and zone 1 in 5 eyes. In 99 eyes (96.1%) neovascularization regressed completely; in 96 eyes (93.2%) the outcome was favorable. In the other 3 eyes, macular dragging formed after regression of neovascularization. Four eyes progressed to more advanced stages of ROP. Repeated laser therapy was performed with the same technique in 10 eyes (9.6%). There was no statistically significant association of unfavorable outcomes with age, birth weight, gestational age, clocks of retinal involvement, and rush disease. No adverse effects of diode laser treatment were observed except for mild injection and edema of conjunctiva in all patients, small conjunctival lacerations in 10 eyes, and small self limited vitreous hemorrhage in 2 eyes. CONCLUSIONS: Transscleral laser treatment may be technically easier for the treatment of retinal periphery, especially in poorly dilating pupils, and obviates the occasional anterior segment complications of the transpupillary approach. This technique can be performed under topical anesthesia. PMID- 20006812 TI - Risk analysis and an alternative protocol for reduction of screening for retinopathy of prematurity. AB - PURPOSE: To determine whether a multivariate risk model can select infants with low-risk eyes for an alternative protocol that reduces retinopathy of prematurity (ROP) screening without loss of effectiveness. METHODS: This was a retrospective, nonrandomized, comparative study. We assigned 712 eyes of 357 premature infants of 401-1,250 g birth weights as high or low risk for prethreshold or threshold ROP using a risk model with variables of birth weight, gestational age, multiple birth, race, and gender. Using simulations, infants with high-risk eyes (p > or = 0.15) were screened conventionally, while those with low-risk eyes were screened with the 35q3 protocol (initial examination at 35 weeks postmenstrual age followed by screening every 3 weeks, with increased or decreased intervals based on ROP severity). The resultant reduction in ROP screening and the delay to detection of severe ROP were calculated. RESULTS: The 35q3 protocol reduced the mean number of eye examinations per infant in the overall population by 13.4% (p = 0.0051). No eyes had a delay to the detection of threshold or type 1 zone 1 ROP. Of a total of 57 type 1 ROP eyes, 2 eyes with type 1 zone 2 ROP were delayed to detection by 1 week. As our study was done prior to the Early Treatment of ROP study, the 2 eyes were observed and regressed without progression to threshold ROP. CONCLUSIONS: A risk-based alternative screening protocol increased the efficiency of ROP screening for infants of 401-1,250 g birth weight without an apparent loss of effectiveness. This investigational approach requires further validation by multicenter studies. PMID- 20006813 TI - Management and outcome of unilateral retinoblastoma. AB - BACKGROUND: Two-thirds of retinoblastoma cases are unilateral, with most presenting with an advanced stage of disease. Primary enucleation is usually the preferred treatment. Conservative therapy may save less involved eyes. METHODS: We retrospectively reviewed the presentation, age at diagnosis, classification, genetic status, treatment, and long-term outcome of 130 patients with unilateral retinoblastoma (1988-2008). RESULTS: The mean age at presentation was 26 months. Based on retinoblastoma gene (RB1) status in tumors, germ-line status was defined in 92% of patients; 13% had a germ-line mutation. The primary treatment of 106 patients was enucleation. Severe disease at presentation (International Intraocular Retinoblastoma Classification [IIRC] group E) was significantly (p < 0.001) associated with adverse histopathological risk factors. Of the 16 patients who underwent eye-conserving therapy, treatment was successful in 9 (IIRC group A, 1; B, 5; C, 3). Two patients with a pertinent family history were diagnosed early and were treated solely with focal therapy. Three patients retained vision of 6/18 or better in the treated eye (median follow-up, 33 months; range, 2-120 months). Seven patients (IIRC group: B, 2; C, 4; D, 1) eventually underwent enucleation. One patient died of metastases following delayed parental consent for enucleation and refusal of prophylactic chemotherapy for high-risk histopathologic features. CONCLUSIONS: Chemotherapy/focal therapy can save selected eyes, but primary enucleation is preferred for advanced unilateral retinoblastoma. "Conservative" treatment is an option when there is good potential for useful vision without prolonged, costly therapy with potential side effects. Simple enucleation reduces the risk of masking high-risk pathology and promotes early return to normal life. PMID- 20006814 TI - Refractive outcomes with secondary intraocular lens implantation in children. AB - PURPOSE: To determine the accuracy of postoperative predicted refractive outcomes in surgically aphakic pediatric patients undergoing secondary intraocular lens (IOL) implantation. Comparisons were also made with other variables historically considered important in cataract surgery. METHODS: Retrospective review of 50 eyes from 35 consecutive pediatric patients (< or =18 years of age) undergoing secondary IOL implantation within the ciliary sulcus or posterior capsular bag. All cases were performed by 1 of 2 surgeons and all refractions were performed manually using the retinoscope by an experienced pediatric ophthalmologist. RESULTS: The mean patient age at the time of the secondary implantation was 6.5 years of age (range, 0.58-15.04). The mean patient age at the time of the primary cataract extraction was 0.78 years (range, 0.08-5.77). For all patients, mean absolute value of prediction error was 1.64 D (SD 1.58 D). There were no significant associations between mean absolute value of prediction error and any of the variables measured including axial length, corneal mean curvature, bag or sulcus implantation, formula used, or age at primary and secondary surgery (p > 0.05). CONCLUSIONS: The mean absolute value of prediction error observed in this study is consistent with previous pediatric primary and secondary IOL data in that it shows a considerable difference from that expected in adult populations. Our findings support the argument that methods currently employed to calculate IOL power may fail to accurately account for all the variations in the eyes of pediatric patients. PMID- 20006815 TI - Theoretical strategy for choosing piggyback intraocular lens powers in young children. AB - INTRODUCTION: The normal growth of a young child's pseudophakic eye can result in a large myopic shift. Temporary polypseudophakia using piggyback intraocular lenses (IOLs) has been proposed as a means to reduce the amount of myopic shift by removing the anterior IOL when the eye becomes sufficiently myopic. Since the rate of refractive growth can be used to predict the refractive curve over time in pseudophakic children, we used this knowledge to develop a theoretical strategy for choosing IOL power combinations for temporary polypseudophakia. METHODS: We used a novel Pediatric Piggyback IOL Calculator to develop a strategy for choosing the powers of the anterior and posterior IOLs. We graphed the predicted results for several combinations of piggyback IOL powers and chose the combination of IOL powers that appeared to give the best results, based on the known rate of refractive growth (5.4 D) and its standard deviation (2.4 D). We aimed for a combination to minimize the hyperopic or myopic refractive error during the first 6 years of life to facilitate amblyopia management and minimize the refractive error at age 20 years. RESULTS: We found optimal results when the initial postoperative goal refraction with polypseudophakia was moderate hyperopia and the anterior IOL had approximately 20% of the total required IOL power. CONCLUSIONS: This theoretical strategy can be used to determine piggyback IOL powers to use in children. PMID- 20006816 TI - Latanoprost in pediatric glaucoma--pediatric exposure over a decade. AB - BACKGROUND: Although numerous studies of latanoprost in adult glaucoma have shown it to be an effective hypotensive agent with a low incidence of side effects, these issues have not been well studied in pediatric glaucomas. The purpose of the current study is to evaluate the safety and intraocular pressure (IOP) lowering effect of latanoprost in various pediatric glaucomas over a long period. SUBJECTS AND METHODS: This retrospective study included all children treated with latanoprost at our institution from 1996 to 2007. Demographic, glaucoma-related, and side-effect information was recorded for each subject. Duration of latanoprost exposure was calculated in child-months (1 child exposed for 1 month). If interpretable IOP data were available, the presence or absence of a treatment response (IOP reduction > or =15% from baseline) was determined for each subject. RESULTS: A total of 115 subjects with latanoprost exposure were identified, with a collective exposure of 2,325 child-months. Exposure for > or =1 year occurred in 52 subjects. Side effects were mild and infrequently reported. Of the 115 subjects, 63 had interpretable IOP data, and 22 (35%) were treatment responders. Predictors of a response included a diagnosis of juvenile open-angle glaucoma, monotherapy, and older age. CONCLUSIONS: This large study of latanoprost-treated children confirms the excellent safety profile of the drug in the treatment of pediatric glaucoma. The study also confirms latanoprost's IOP lowering ability in older children with juvenile open-angle glaucoma and in some children with aphakic glaucoma. Prospective studies are needed to better define the optimal role of latanoprost in the treatment of pediatric glaucoma, especially congenital glaucoma. PMID- 20006817 TI - Spectral domain optical coherence tomography for detection of foveal morphology in patients with nystagmus. AB - PURPOSE: To evaluate the feasibility of spectral domain optical coherence tomography (SD-OCT) macular scanning as a means of studying the afferent visual system in nystagmus patients. METHODS: Nystagmus patients who underwent SD-OCT, clinical evaluation, and eye movement recordings were recruited for this prospective, single-center, noncomparative study. Three SD-OCT macular three dimensional cube scans per eye (200 x 200 x 1024 samplings in a 6 x 6 mm region) were obtained for qualitative retinal morphology analysis. RESULTS: Nineteen patients (6-68 years; average, 19 years) were analyzed. Of these, 17 patients had infantile nystagmus syndrome, and 2 had fusion maldevelopment nystagmus; 17 patients (89%) had associated sensory system abnormalities, including 9 (47%) with albinism. Macular images were successfully obtained in all but 1 patient (95%). Of the 8 successfully imaged oculocutaneous patients, 7 patients demonstrated "fovea plana," and all demonstrated abnormal morphology. CONCLUSION: SD-OCT reliably provides detailed structural imaging of the fovea in nystagmus patients. PMID- 20006818 TI - Relationship of infantile periocular hemangioma depth to growth and regression pattern. AB - PURPOSE: Most infantile periocular hemangiomas undergo rapid growth in the first year of life, followed by gradual resolution over years. Treatment is indicated if vision is compromised and is usually continued through the growth phase. The objective of this study was to determine which clinical characteristics might aid in the prediction of growth and/or regression patterns of periocular hemangiomas. METHODS: Retrospective review of medical records and photographs of children with periocular hemangiomas presenting to a UK pediatric eye unit over a 7-year period. Age at presentation, growth pattern, size, location, amblyopia, and refractive status were documented. RESULTS: Forty-two infants with periocular hemangiomas were evaluated between 2000 and 2007, with a mean follow-up of 24 months (range, 6 months to 5 years). One-third (n=14, 33%) of the hemangiomas were superficial (strawberry nevi); one-third were subcutaneous (n=13, 31%), and the remainder were mixed (n=8, 19%) and orbital (n=7, 17%). There was a marked difference between the growth patterns of superficial (strawberry nevi) and deeper hemangiomas (orbital and subcutaneous), with a more prolonged period of growth noted in the deeper hemangiomas. CONCLUSIONS: Periocular hemangiomas with a deep component tend to have a later onset and prolonged period of growth compared to strawberry nevi. Clinically evident depth of the hemangioma appears to be a valuable predictor of rapidity of resolution. This finding may be useful in assessing prognosis and planning treatment of infantile periocular hemangiomas. PMID- 20006819 TI - Frequency of the superior rectus muscle overaction/contracture syndrome in unilateral fourth nerve palsy. AB - INTRODUCTION: Superior oblique palsy is accompanied in most cases by overaction of the muscle's ipsilateral antagonist, the inferior oblique muscle. Overaction and contracture of the ipsilateral superior rectus muscle in patients with unilateral fourth (trochlear) nerve palsy is seldom discussed in the literature. The purpose of this study is to evaluate the frequency of superior rectus muscle overaction/contracture syndrome in patients with unilateral trochlear nerve palsy. SUBJECTS AND METHODS: The records of 198 patients with unilateral trochlear nerve palsy examined by the authors between July 1987 and July 2008 were reviewed retrospectively. All patients underwent complete eye examination with measurement of the deviation in the 9 positions of gaze and with the head tilted to both sides in all cooperative patients. Selection criteria for superior rectus muscle overaction/contracture syndrome in these patients were as follows: vertical deviation of 15(Delta) or larger in primary position, equal or larger hypertropia with the ipsilateral forced tilt test than with the eyes looking straight ahead, more than 5(Delta) hypertropia of the affected eye in horizontal gaze to the same side, hypertropia in all upgazes, and overaction of the contralateral superior oblique muscle. RESULTS: Of 198 patients, 33 (16.6%) met the selection criteria for superior rectus muscle overaction/contracture syndrome. CONCLUSIONS: Superior rectus muscle overaction/contracture syndrome frequently occurs in unilateral superior oblique palsy. PMID- 20006820 TI - Effects of vertical offsets of the horizontal rectus muscles in V-pattern exotropia without oblique dysfunction. AB - PURPOSE: To investigate the effects of vertical offsets of the horizontal rectus muscle in V-pattern exotropia without oblique dysfunction. METHODS: A computerized patient database was searched for patients treated with full tendon offsets for V-pattern strabismus from 2002 to 2007. Patients with clinically significant inferior oblique overaction were excluded. Surgery included upward transposition of lateral rectus muscles and/or downward transposition of medial rectus muscles. Medical records were retrospectively analyzed for alignment in the primary position, 30 degrees upgaze, and 30 degrees downgaze before and 3 months after surgery. The main outcome measure was the change in V pattern after surgery. RESULTS: The search identified 11 patients who met inclusion criteria. Of these, 6 underwent bilateral lateral rectus recessions, 1 had 3-muscle surgery, and 4 had monocular recession-resection surgery. The preoperative V pattern (horizontal) deviation between upgaze and downgaze was 22.2(Delta)+/ 5.0(Delta). Postoperative deviation at 3 months was 3.5(Delta)+/-3.9(Delta), a decrease of 18.9(Delta)+/-7.5(Delta). Postoperatively, no patient had an A pattern deviation or subjective torsional disturbance. The amount of V-pattern correction was closely correlated with the size of the preoperative V pattern (r=0.84). CONCLUSIONS: In patients with V-pattern exotropia without inferior oblique dysfunction, vertical offsets of the horizontal rectus muscles effectively corrected the pattern deviation. The amount of correction was closely correlated with the size of the preoperative V pattern. PMID- 20006822 TI - Duration of conjunctival redness following adult strabismus surgery. AB - PURPOSE: To determine the duration of postoperative conjunctival injection following strabismus surgery and to assess how this is affected by previous extraocular muscle surgery. This would improve preoperative counseling of strabismus patients. METHODS: Subjective evaluation of conjunctival redness based on patient questionnaire response. RESULTS: Fifty-three patients returned completed questionnaires. A total of 93 muscles were operated on. Of these, 46 had not undergone previous operations; 47 had. Previously unoperated eyes remained red for a median duration of 9.5 weeks; reoperated eyes remained red for a median of 11 weeks. Of the 93 muscles, 50 were sutured with adjustable sutures. These remained red for a median duration of 11 weeks. Muscles tied with nonadjustable sutures remained red for a median of 10 weeks. CONCLUSIONS: In both previously unoperated and reoperated eyes, conjunctival redness resolved in approximately 10 weeks; adjustable sutures did not alter the duration of redness significantly. PMID- 20006821 TI - Precaruncular approach for medial orbital wall periosteal anchoring of the globe in oculomotor nerve palsy. AB - PURPOSE: To evaluate a precaruncular approach for fixation of the globe to the medial orbital wall periosteum for management of complete third (oculomotor) nerve palsy. METHODS: Consecutive patients with severe unilateral oculomotor nerve palsy present for at least 2 years were prospectively treated and evaluated. In all patients, a 12-16 mm recession of the lateral rectus muscle was performed along with the precaruncular anchoring procedure. Follow-up evaluations were performed at 1 week, and at 1, 2, and 3 months after surgery, with ongoing follow-up at 3 month intervals. RESULTS: Fourteen eyes of 14 patients with complete oculomotor nerve palsy were included in the series. The median horizontal preoperative deviation of -90(Delta) +/- 4.8(Delta) reduced to 10(Delta) +/- 8.3(Delta). The vertical deviation reduced from 24(Delta) +/- 7.4(Delta) to 12.8(Delta) +/- 6.0(Delta). Mean follow-up was 8.9 +/- 5.5 months (range, 6-21 months). A slight exotropic drift was observed over 4 to 6 weeks following surgery in all cases. Satisfactory alignment was observed in 13 of the 14 cases (92.85%) over the duration of the follow-up period. CONCLUSIONS: Anchoring the globe to the medial orbital wall using a precaruncular approach is a viable option in the management of complete external oculomotor nerve palsy. PMID- 20006823 TI - Leber congenital amaurosis: clinical correlations with genotypes, gene therapy trials update, and future directions. AB - Leber congenital amaurosis comprises a group of early onset childhood retinal dystrophies, characterized by vision loss, nystagmus, and severe retinal dysfunction. To date, 15 causative genes have been identified that account for the heterogeneous presentation and clinical course. Knowledge of key aspects of phenotype and clinical course can contribute to the determination of a precise genetic etiology using genetic testing. Gene-based therapies are emerging, and knowledge of a patient's genotype is essential. A review of clinical presentation and disease course, their correlation to specific genotypes, and underlying physiological mechanisms, coupled with the latest results of human gene therapy trials, will assist the clinician in patient diagnosis and counseling. PMID- 20006824 TI - Photographic assessment of changes in torsional strabismus. AB - The horizontal and vertical components of strabismus are measured routinely and relatively easily in the clinical setting using prism-and-cover and/or corneal light reflex tests. The third dimension of ocular alignment, ocular torsion, is more difficult to assess. Objective torsional deviation (cyclotropia) is evaluated qualitatively with fundus examination. For quantitative assessment, however, fundus photography is needed, which may not always be available during a strabismus examination and typically requires pupil dilation. We present a simple, inexpensive photographic technique to assess changes in iris torsion and evaluate its accuracy by comparison with fundus photography. Using a consumer grade digital camera, basic photographic editing software, and a data worksheet, this technique assesses changes in objective torsion with accuracy and retest variability of both approximately 1 degrees. PMID- 20006825 TI - Transient internal ophthalmoplegia after inferior oblique myectomy. AB - Internal ophthalmoplegia causing pupillary dilatation and loss of accommodation following damage to the ciliary ganglion is a rare complication of strabismus surgery. Here we report a case of parasympathetic neuropraxia resulting in transient internal ophthalmoplegia after inferior oblique myectomy in a 12-year old girl. Short-term symptomatic relief was achieved with 1% pilocarpine. Normal visual function returned over several months. PMID- 20006826 TI - A case of spina bifida with ocular manifestations. AB - Neural tube defects are common congenital abnormalities associated with a variety of complications. Spina bifida, among the most common, results from failure of the neural tube and the vertebral bones to close during embryogenesis. Myelomeningocele is the most common severe form of spina bifida. It may be associated with various anomalies in different tissues. Here we report a case of spina bifida with associated ocular anomalies and review the literature of previous spina bifida cases with ocular manifestations. PMID- 20006827 TI - Bartonella henselae neuroretinitis in a 15-year-old girl with chronic myelogenous leukemia. AB - A 15-year-old girl being treated with imatinib for chronic myelogenous leukemia (CML) presented with acute vision loss in her right eye accompanied by swelling of the right side of her neck. On examination, she was found to have disk and macular edema of the right eye. Fine-needle aspiration and excisional biopsies of an enlarged submandibular lymph node were negative for malignancy. Although initial serologies were equivocal for Bartonella henselae, repeat serologies performed 1 week later upon the appearance of a macular star were positive. This is the first reported case of B. henselae in a patient with CML. PMID- 20006829 TI - Pupillary light reflexes in premature infants prior to 30 weeks postmenstrual age. AB - Data regarding the pupillary responses in very premature neonates is scarce; what data exist, moreover, is not recent. The purpose of this pilot study is to collect data on direct and consensual pupillary light responses before 30 weeks postmenstrual age. Six neonates were studied. Mean pupillary size at rest was 3.6 +/- 0.4 mm. No direct or consensual responses to light were present in any of 12 eyes. Accurate information about pupillary reflexes in very premature neonates provides relevant information about the development of the visual and neurologic systems. Available information about the development of the Edinger-Westphal nucleus, sphincter pupillary muscle, optic nerve myelinization, and autonomic nervous system is briefly reviewed. PMID- 20006828 TI - Orbital teratoma: case report and management review. AB - We present a case of benign mature orbital teratoma in a neonatal boy that involved the chiasmal area and cavernous sinus and displaced the nasopharynx. Visual potential was deemed to be good. Because of the chiasmal involvement, complete surgical resection, which would have compromised his visual function, was considered inadvisable. A debulking procedure, mainly of the cystic parts of the tumor, was performed. The patient remains stable at 3 years of age, with no further growth of the tumor and good vision in the affected eye. PMID- 20006830 TI - Ocular albinism with absent foveal pits but without nystagmus, photophobia, or severely reduced vision. AB - A 9-year-old Caucasian girl of northern European ancestry presented with findings suggestive of ocular albinism, although she maintains good visual acuity and lacks nystagmus and photophobia. DNA analysis revealed that the patient is a compound heterozygote for mutations in the tyrosinase gene, which is typically associated with overt, generalized oculocutaneous albinism and severe ocular symptoms. Her particular genotype confers no apparent cutaneous disease and only mild ocular features. PMID- 20006831 TI - A case of intraocular yolk sac tumor in a child and its pathogenesis. AB - While yolk sac tumor is one of the most common malignant germ cell tumors occurring in young children, it is rarely found in extragonadal sites. We report a case of intraocular yolk sac tumor in a 4-year-old boy. The diagnosis was confirmed by histologic examination and by the rapid normalization of serum alpha fetoprotein level following enucleation. We propose that yolk sac cells can potentially migrate into the eye at 22 days of embryonic life during neural tube formation, when the head and tail of the neuropore open contemporaneously and communicate with the amniotic cavity. PMID- 20006833 TI - Surgical technique for removing congenital fibrovascular pupillary membrane, with clinicopathological correlation. AB - Congenital fibrovascular pupillary membrane, rare in newborns, poses challenges for the ophthalmic surgeon. Predicting its clinical course is difficult, as is removing the membrane surgically without inducing a cataract. We describe a 22 day-old girl who presented with a pupillary membrane causing progressive decrease in the size of the dilated pupil, necessitating removal at 4 months of age. Histopathology demonstrated tissue of embryonic muscle origin, suggesting that the membrane was more likely a remnant of fetal iris than of fetal vasculature. The smooth muscle or fibroblastic elements in the tissue explains progressive contracture of the membrane. PMID- 20006832 TI - Ranibizumab for coloboma-related choroidal neovascular membrane in a child. AB - Optic nerve and retinochoroidal coloboma are caused by incomplete closure of the embryonic fissure during fetal development.(1) Affected patients carry a risk for retinal detachment and less so for choroidal neovascular membranes (CNVM) secondary to the altered anatomy. Because of the rarity of this condition and the even more unusual occurrence of such complications, there are only a few case reports on the treatment of coloboma-related CNVM. Herein we report the results of ranibizumab and laser photocoagulation for coloboma-related CNVM in a child. PMID- 20006834 TI - Recurrent corneal metallic foreign bodies in children with autism spectrum disorders. AB - Corneal metallic foreign body is a common condition in working-age adults due to occupational exposure, such as grinding, welding, and hammering. Ocular trauma is a leading cause of vision loss in children; however, corneal metallic foreign bodies are rare in the pediatric population. Here we describe 2 children suffering from autism spectrum disorders presenting with recurrent corneal metallic foreign bodies. Meticulous history revealed that both children extensively used a therapeutic home swing with metallic suspensions, suggesting the potential mechanism for the recurrent corneal metallic foreign bodies. Furthermore, the use of protective eyewear during swinging prevented further recurrences. Awareness of this potential danger is particularly important in children with disabilities, who may have difficulty communicating their concerns and cooperating during examination and treatment. PMID- 20006835 TI - Regulatory natural killer cells: new players in the IL-10 anti-inflammatory response. AB - Natural Killer (NK) cells boost immune defense via their cytolytic activity and their capacity to produce interferon-gamma. Now Perona-Wright et al. (2009), in this issue of Cell Host & Microbe, show that NK cells can also dampen immune responses to diverse pathogens via secretion of the anti-inflammatory cytokine interleukin-10. Remarkably, this immunosuppressive activity only occurs during systemic infections. PMID- 20006836 TI - Hepatitis C and evasion of the interferon system: a PKR paradigm. AB - Hepatitis C virus (HCV) is resistant to the antiviral cytokine type I interferon, representing a major clinical problem. Garaigorta and Chisari (2009) reveal that HCV uses the activation of the ds-RNA-dependent protein kinase R, which phosphorylates and inhibits the translation initiation factor eIF-2 alpha, to block translation of interferon-stimulated genes. PMID- 20006837 TI - Poxviruses taking a ride on actin: new users of known hardware. AB - Vaccinia virus uses A36 to recruit the actin polymerization effectors Nck and N WASP to drive actin tail formation. Now, Dodding and Way identify functional orthologs of A36 in other vertebrate poxviruses that harbor varying numbers of Nck-binding sites and can substitute for A36 despite no sequence homology. PMID- 20006838 TI - Malaria parasites do the stick-and-slip shuffle. AB - Munter et al. (2009) demonstrate that malaria parasite movement is highly complex, involving cycles of fast sprints and slow glides over their substrate. This "stick-and-slip shuffle" is controlled by the parasite's secreted adhesive proteins and their interaction with the dynamic actin cytoskeleton. PMID- 20006839 TI - Systemic but not local infections elicit immunosuppressive IL-10 production by natural killer cells. AB - Surviving infection represents a balance between the proinflammatory responses needed to eliminate the pathogen, and anti-inflammatory signals limiting damage to the host. IL-10 is a potent immunosuppressive cytokine whose impact is determined by the timing and localization of release. We show that NK cells rapidly express IL-10 during acute infection with diverse rapidly disseminating pathogens. The proinflammatory cytokine IL-12 was necessary and sufficient for NK cell induction of IL-10. NK cells from mice with systemic parasitic infection inhibited dendritic cell release of IL-12 in an IL-10-dependent manner, and NK cell depletion resulted in elevated serum IL-12. These data suggest an innate, negative feedback loop in which IL-12 limits its own production by eliciting IL 10 from NK cells. In contrast to disseminating pathogens, locally restricted infections did not elicit NK cell IL-10. Thus systemic infections uniquely engage NK cells in an IL-10-mediated immunoregulatory circuit that functions to alleviate inflammation. PMID- 20006840 TI - Hepatitis C virus blocks interferon effector function by inducing protein kinase R phosphorylation. AB - Hepatitis C virus (HCV) is a single-stranded RNA virus encoding a single polyprotein whose translation is driven by an internal ribosome entry site (IRES). HCV infection strongly induces antiviral interferon-stimulated gene (ISG) expression in the liver, yet it persists, suggesting that HCV can block ISG effector function. We now show that HCV infection triggers phosphorylation and activation of the RNA-dependent protein kinase PKR, which inhibits eukaryotic translation initiation factor eIF2 alpha and attenuates ISG protein expression despite normal ISG mRNA induction. ISG protein induction is restored and the antiviral effects of interferon are enhanced when PKR expression is suppressed in interferon-treated infected cells. Whereas host protein translation, including antiviral ISGs, is suppressed by activated PKR, HCV IRES-dependent translation is not. These results suggest that the ability of HCV to activate PKR may, paradoxically, be advantageous for the virus during an IFN response by preferentially suppressing the translation of ISGs. PMID- 20006841 TI - Adenovirus transport via direct interaction of cytoplasmic dynein with the viral capsid hexon subunit. AB - Early in infection, adenovirus travels to the nucleus as a naked capsid using the microtubule motor cytoplasmic dynein. How the dynein complex is recruited to viral cargo remains unclear. We find that cytoplasmic dynein and its associated proteins dynactin and NudE/NudEL, but not LIS1 or ZW10, colocalized with incoming, postendosomal adenovirus particles. However, in contrast to physiological cargos, dynein binding to adenovirus was independent of these dynein-associated proteins. Dynein itself directly interacted through its intermediate and light intermediate chains with the adenovirus capsid subunit hexon in a pH-dependent manner. Expression of hexon or injection of anti-hexon antibody inhibited virus transport but not physiological dynein function. These results identify hexon as a direct receptor for cytoplasmic dynein and demonstrate that hexon recruits dynein for transport to the nucleus by a mechanism distinct from that for physiological dynein cargo. PMID- 20006842 TI - Nck- and N-WASP-dependent actin-based motility is conserved in divergent vertebrate poxviruses. AB - Vaccinia virus enhances its cell-to-cell spread by stimulating actin polymerization via Src- and Abl-mediated phosphorylation of the highly conserved orthopoxvirus protein A36. The Yatapoxvirus, Yaba-like disease virus (YLDV), also induces actin polymerization, although it lacks an obvious A36 ortholog. We found that the YLDV protein YL126 can functionally replace A36 to promote Nck- and N WASP-dependent actin polymerization. At least five phosphorylated tyrosines in YL126, rather than a single residue as in A36, are able to recruit Nck to promote actin polymerization. As is the case for A36, YL126-mediated actin tail formation is enhanced by the recruitment of Grb2 via a single phosphorylated tyrosine in YL126. Furthermore, highly divergent YL126 orthologs in Yaba monkey tumor, lumpy skin disease, Shope fibroma, myxoma, and swine and squirrel poxviruses also stimulate Nck- and N-WASP-dependent actin polymerization, suggesting that actin based motility represents a common mechanism to enhance the cell-to-cell spread of vertebrate poxviruses. PMID- 20006843 TI - Plasmodium sporozoite motility is modulated by the turnover of discrete adhesion sites. AB - Sporozoites are the highly motile stages of the malaria parasite injected into the host's skin during a mosquito bite. In order to navigate inside of the host, sporozoites rely on actin-dependent gliding motility. Although the major components of the gliding machinery are known, the spatiotemporal dynamics of the proteins and the underlying mechanism powering forward locomotion remain unclear. Here, we show that sporozoite motility is characterized by a continuous sequence of stick-and-slip phases. Reflection interference contrast and traction force microscopy identified the repeated turnover of discrete adhesion sites as the underlying mechanism of this substrate-dependent type of motility. Transient forces correlated with the formation and rupture of distinct substrate contact sites and were dependent on actin dynamics. Further, we show that the essential sporozoite surface protein TRAP is critical for the regulated formation and rupture of adhesion sites but is dispensable for retrograde capping. PMID- 20006844 TI - Cellular DNA ligase I is recruited to cytoplasmic vaccinia virus factories and masks the role of the vaccinia ligase in viral DNA replication. AB - Vaccinia virus (VACV) encodes DNA polymerase and additional proteins that enable cytoplasmic replication. We confirmed the ability of VACV DNA ligase mutants to replicate and tested the hypothesis that cellular ligases compensate for loss of viral gene expression. RNA silencing of human DNA ligase I expression and a small molecule inhibitor of human DNA ligase I [corrected] severely reduced replication of viral DNA in cells infected with VACV ligase-deficient mutants, indicating that the cellular enzyme plays a complementary role. Replication of ligase deficient VACV was greatly reduced and delayed in resting primary cells, correlating with initial low levels of ligase I and subsequent viral induction and localization of ligase I in virus factories. These studies indicate that DNA ligation is essential for poxvirus replication and explain the ability of ligase deletion mutants to replicate in dividing cells but exhibit decreased pathogenicity in mice. Encoding its own ligase might allow VACV to "jump-start" DNA synthesis. PMID- 20006845 TI - Regulation of KSHV lytic switch protein expression by a virus-encoded microRNA: an evolutionary adaptation that fine-tunes lytic reactivation. AB - Herpesviruses encode numerous microRNAs (miRNAs), most of whose functions are unknown. The Kaposi's sarcoma-associated herpesvirus (KSHV) encodes 17 known miRNAs as part of its latency program, suggesting that these RNAs might function to regulate the latent state. Here we show that one of these KSHV miRNAs, miRK9( *), targets a sequence in the 3' untranslated region (UTR) of the mRNA encoding the major lytic switch protein (RTA), which controls viral reactivation from latency. Ectopic expression of miRK9( *) impairs RTA synthesis, while its specific antagonism in latently infected cells enhances spontaneous lytic reactivation frequency by 2- to 3-fold. Mutation of the recognition sequence in the RTA 3'UTR abolishes RTA downregulation by miRK9( *). We propose that miRNA targeting of RTA, while not the primary regulator of the lytic switch, functions like a safety mechanism on the trigger of lytic reactivation, preventing stochastic variations in basal RTA transcription from activating inappropriate entry into the lytic cycle. PMID- 20006846 TI - Prevalence and characteristics of female sexual dysfunction in a sample of women from Upper Egypt. AB - OBJECTIVE: To study the prevalence and associated risk factors for female sexual dysfunction (FSD) in Upper Egypt. METHODS: Married women attending the outpatient clinic of Sohag University Hospital between February 2008 and March 2009 were recruited. Through direct interviews, each woman completed a questionnaire that included questions on sexual dysfunction. RESULTS: From 648 recruited patients, 47 declined to participate in the study. Of the 601 remaining participants, 462 women (76.9%) reported 1 or more sexual problems. Low sexual desire was the most common sexual problem (66.4%). Patients with FSD were significantly more likely to be older than 40 years, have sexual intercourse fewer than 3 times a week, have been married for 10 years or more, have 5 children or more, be circumcised, have a husband aged 40 years or more, and be postmenopausal. Age of the women maintained a statistically significant positive relationship with FSD in the regression model (odds ratio 1.39; 95% CI, 1.26-1.53). CONCLUSIONS: There was a high prevalence of FSD in this sample of women from Upper Egypt. The prevalence of sexual problems increased with increasing age of the women. PMID- 20006847 TI - Hierarchically imprinted polymer substrates for enhanced attachment of Escherichia coli. AB - Escherichia coli (E. coli) detection is important for ensuring human health and public security. One critical step in most detection methods is to have the E. coli cells attach to the substrate or transducer of a biosensor before they can be detected and/or identified. In this context, a chemical or physical enhancement effect arising from the substrate will help to achieve a high sensitivity of bacterial detection. This work makes use of hierarchically imprinted surface structures to demonstrate such effect using quartz crystal microbalance (QCM). Specifically, hierarchical structures are imprinted on polystyrene coated resonance crystals of QCM; such crystals, after incubation in an E. coli suspension of reduced concentration (1x10(4) colony forming units/mL), exhibit improved resonance frequency shifts, which are 1-2 orders of magnitude higher than those without the hierarchical structures. The enhancement effect is attributed to the enlarged surface area of the substrate and the way it immobilizes the bacteria. As revealed by scanning electron microscopy, the hierarchical substrates immobilize the E. coli cells by both trapping them in the micro-trenches and having them adhere to the nano-protrusions, while the single level imprinted structures accommodate the cells mainly in the trenches or over the protrusions, instead of both. PMID- 20006848 TI - Control of the morphology and size of magnetite particles with peptides mimicking the Mms6 protein from magnetotactic bacteria. AB - Mms6 is a dominant protein that tightly associates with the surface of bacterial magnetites in Magnetospirillum magneticum AMB-1. The protein has previously been shown to mediate the formation of uniform magnetite crystals of cubo-octahedral morphology consisting of (1 1 1) and (1 0 0) crystal faces with a narrow size distribution during chemical magnetite synthesis. In order to understand the role of this protein in chemical magnetite synthesis, magnetite formation was investigated using synthetic peptides mimicking the Mms6 protein. Particles that were synthesized in the presence of short peptides harbouring the C-terminal acidic region of Mms6 exhibited a spherical morphology with circularities of 0.70 0.90 similar to those of bacterial magnetites and particles formed in the presence of the Mms6 protein. In contrast, a rectangular morphology with circularities of 0.60-0.85 were obtained when other peptides were used for synthesis. The results indicated that the C-terminal region of the Mms6 protein has significant control over the morphology of magnetite crystals in the chemical synthetic method. This method can, therefore, be useful as an alternative method of controlling the size and morphology of magnetite crystals under ambient conditions. PMID- 20006849 TI - Further support for the role of dysfunctional attitudes in models of real-world functioning in schizophrenia. AB - According to A.T. Beck and colleagues' cognitive formulation of poor functioning in schizophrenia, maladaptive cognitive appraisals play a key role in the expression and persistence of negative symptoms and associated real-world functioning deficits. They provided initial support for this model by showing that dysfunctional attitudes are elevated in schizophrenia and account for significant variance in negative symptoms and subjective quality of life. The current study used structural equation modeling to further evaluate the contribution of dysfunctional attitudes to outcome in schizophrenia. One hundred eleven outpatients and 67 healthy controls completed a Dysfunctional Attitudes Scale, and patients completed a competence measure of functional capacity, clinical ratings of negative symptoms, and interview-based ratings of real-world functioning. Patients reported higher defeatist performance beliefs than controls and these were significantly related to lower functional capacity, higher negative symptoms, and worse community functioning. Consistent with Beck and colleagues' formulation, modeling analyses indicated a significant indirect pathway from functional capacity-->dysfunctional attitudes-->negative symptoms- >real-world functioning. These findings support the value of dysfunctional attitudes for understanding the determinants of outcome in schizophrenia and suggest that therapeutic interventions targeting these attitudes may facilitate functional recovery. PMID- 20006850 TI - Siblings with the adult-onset slowly progressive type of pantothenate kinase associated neurodegeneration and a novel mutation, Ile346Ser, in PANK2: clinical features and (99m)Tc-ECD brain perfusion SPECT findings. AB - Pantothenate kinase-associated neurodegeneration (PKAN), formerly known as Hallervorden-Spatz syndrome (HSS), is an autosomal recessive neurodegenerative disorder characterized by iron accumulation in the brain. Mutations in the pantothenate kinase 2 (PANK2) gene are known to be responsible for PKAN. Several studies have revealed correlations between clinical phenotypes and particular PANK2 mutations. The adult-onset slowly progressive type of PKAN with PANK2 mutations is very rare. In this report, we describe siblings with the adult-onset slowly progressive type of PKAN with a novel mutation, Ile346Ser, in PANK2. The siblings had the same mutation in PANK2 and had common clinical signs such as misalignment of teeth, a high arched palate, hollow feet, a slight cognitive decline, and an apparent executive dysfunction, although they showed different patterns of movement disorders. Thus, even if PKAN patients have identical mutations, it is likely that they will present with different types of movement disorders. Brain perfusion single photon emission computed tomography in both patients showed decreased regional cerebral blood flow in the bilateral frontoparietal lobes, the globus pallidus, the striatum, and around the ventriculus quartus. Cardiac uptake of [(123)I] meta-iodobenzylguanidine was normal in both patients. Analysis of genotype-phenotype correlations and the elucidation of mutational effects on pantothenate kinase 2 function, expression, and structure are important for understanding the mechanisms of PKAN. PMID- 20006851 TI - Prognostic factors of papillary renal cell carcinoma: results from a multi institutional series after pathological review. AB - PURPOSE: We examined papillary renal cell carcinoma prognostic variables and validated the 2002 UICC TNM staging system in a multicenter analysis. MATERIALS AND METHODS: From 10 urological institutions in Germany followup data were collected on a total of 675 patients with papillary renal cell carcinoma. Central pathological review was done to validate external histopathological diagnoses. The Kaplan-Meier method was used to derive cumulative cancer specific and overall survival, and the log rank test was used to compare the curves of 2 or more groups. For multivariate analysis of prognostic factors Cox regression analysis was done. All proportional hazard assumptions were systemically verified using the Grambsch-Therneau test. RESULTS: Cancer specific survival was significantly related to TNM stage and histological grading on univariate and multivariate analyses. Five-year cancer specific survival in pT1b cases was significantly shorter than in pT1a cases (90.0% vs 98.3%, p = 0.017). No significant difference was found between pT1b and pT2 tumors. Patients with pT3 or greater disease were at high risk for metastasis (50.6%) while metastatic disease associated with pT2 or less tumors occurred in 7.8% (p <0.0001). After metastatic disease was present the prognosis was poor with 7.2% 5-year cancer specific survival. Age was associated with poor prognosis in the subgroup with pT3 or greater tumors on univariate analysis (p = 0.026) but not on multivariate analysis. CONCLUSIONS: In its current form the 2002 UICC TNM staging system is not applicable to papillary renal cell carcinoma. Clinical and radiological followup should be offered at frequent intervals to patients with venous thrombus and/or locally advanced disease. The role of age remains unclear but should not be underestimated in risk stratification after surgery. PMID- 20006852 TI - Effect of reclassification on the incidence of benign and malignant renal tumors. AB - PURPOSE: The incidence of benign renal tumors has increased in recent years. This trend is commonly attributed to the increased use of cross-sectional imaging and minimally invasive surgical approaches. An alternative hypothesis is that recent changes in histological classification are responsible for the increasing incidence. To further investigate the impact of histological reclassification we reexamined all excised renal masses using the 2004 WHO criteria and compared this histological classification to the prior criteria. MATERIALS AND METHODS: We identified 1,101 consecutive partial and radical nephrectomy cases managed at our institution from 1989 to 2003. All histopathological sections were rereviewed by a single pathologist and reclassified according to 2004 WHO criteria. The percentages of benign lesions per year according to the prior histological and current WHO 2004 histological criteria were compared. RESULTS: Of the 1,101 renal masses 132 (12.0%) and 165 (15.0%) were classified as benign using prior and current WHO criteria, respectively. On average the WHO criteria diagnosed more benign tumors per year than the prior criteria (p = 0.004). Linear regression demonstrated a similar, persistent increase in benign diagnoses per year of 0.69% (WHO) and 1.22% (prior) during the 14-year period (p = 0.33). All masses reclassified as benign were oncocytoma (33). CONCLUSIONS: Implementation of the 2004 WHO criteria is contributing to the increase in diagnosis of benign renal lesions, specifically oncocytoma. Changes in histological classification do not account for the entire increase. Other factors, which remain to be delineated, are also contributing to the increase in the diagnosis of benign renal lesions. PMID- 20006853 TI - Editorial comment. PMID- 20006854 TI - Impact of hormonal therapy on intermediate risk prostate cancer treated with combination brachytherapy and external beam irradiation. AB - PURPOSE: We assessed the impact of androgen suppressive therapy on biochemical failure in patients with intermediate risk prostate cancer treated with brachytherapy and external beam irradiation. MATERIALS AND METHODS: From 1994 to 2006, 432 patients with intermediate risk prostate cancer as defined by the National Comprehensive Cancer Network were treated with low dose rate brachytherapy and external beam irradiation with or without 9 months of androgen suppressive therapy. Gleason score was 7 in 76% of cases and prostate specific antigen was 1.4 to 20 ng/ml (median 7.6). Of the patients 350 received androgen suppressive therapy and 82 did not. The biologically effective dose was 142 to 280 Gy2 (median 206). Followup was 23 to 155 months (median 56). RESULTS: The overall 8-year biochemical failure-free rate using the Phoenix definition in patients with vs without androgen suppressive therapy was 92% vs 92% (p = 0.4). The therapy had no significant impact on the biochemical failure-free rate in patients with Gleason score 7 (92% vs 90.5%, p = 0.55), prostate specific antigen 10 to 20 ng/ml (92% vs 100%, p = 0.32), T2b-T2c disease (89.5% vs 97%, p = 0.27) and more than 1 intermediate risk feature (90% vs 100%, p = 0.2). CONCLUSIONS: We addressed the relative importance of radiation dose vs hormonal therapy for intermediate risk prostate cancer. With high biologically effective dose combination treatment androgen suppressive therapy did not have a significant impact on the 8-year biochemical failure-free rate. We question its routine use in this setting. PMID- 20006855 TI - Editorial comment. PMID- 20006856 TI - How do commonly performed lymphadenectomy templates influence bladder cancer nodal stage? AB - PURPOSE: Determining pathological nodal stage in patients with bladder cancer is important for prognosis. We determined how the extent of lymphadenectomy and the lymph node count influence accurate nodal staging. MATERIALS AND METHODS: The study included 120 patients who underwent at least extended lymphadenectomy at radical cystectomy. Different anatomical templates for lymphadenectomy were evaluated for nodal staging accuracy. The cumulative percent was plotted to determine a lymph node count that confidently identified node positive cases. RESULTS: The mean +/- SD total lymph node count in the study population was 36.9 +/- 14.8 at extended lymphadenectomy. Of the patients 36 (30%) had lymph node metastasis, including 14 (39%) with metastasis involving the common iliac and/or presacral lymph nodes. Limited, standard and extended lymphadenectomy accurately identified 75%, 88.9% and 100% of node positive cases, respectively. Removing 23 and 27 lymph nodes provided 80% and 90% confidence, respectively, that a case was accurately staged as pN0. No patient had lymph node metastasis above the aortic bifurcation without nodal metastasis below the aortic bifurcation and none had a change in pN stage by extending lymphadenectomy above the aortic bifurcation. CONCLUSIONS: To accurately identify node positive and negative cases, and correctly assign pN stage in node positive cases it is necessary to perform extended lymphadenectomy. Identifying at least 23 to 27 lymph nodes on final pathological evaluation provides a high level of confidence that a case is correctly staged as node positive or negative. PMID- 20006857 TI - New tools to study bladder dysfunction. PMID- 20006858 TI - Expression of HYAL1 and survivin RNA as diagnostic molecular markers for bladder cancer. AB - PURPOSE: Urinary tumor markers that help in the early detection of bladder cancer promise a significant improvement in sensitivity, specificity and convenience over conventional, invasive diagnostic tests. We assessed the diagnostic efficacy of hyaluronidase (HYAL1) and survivin for early bladder cancer detection. MATERIALS AND METHODS: The study included 166 patients diagnosed with bladder carcinoma, 112 with benign bladder lesions and 100 healthy volunteers who served as controls. All underwent serological assessment of schistosomiasis antibody, urine cytology, and hyaluronidase (HYAL1) and survivin RNA estimation by qualitative and semiquantitative reverse transcriptase-polymerase chain reaction in urothelial cells from voided urine. RESULTS: Positivity rates of HYAL1 RNA and survivin RNA on qualitative reverse transcriptase-polymerase chain reaction were significantly different among the 3 groups. Mean rank using semiquantitative method was increased in the malignant vs the other groups. The best cutoff for HYAL1 and survivin RNA was 0.25 each. Using these cutoffs HYAL1 and survivin RNA sensitivity was 91% and 75%, respectively, with absolute specificity. HYAL1 RNA detected all patients with stages 0 and I bladder cancer (p <0.037). Urine cytology sensitivity improved when combined with hyaluronidase or survivin RNA on semiquantitative reverse transcriptase-polymerase chain reaction. CONCLUSIONS: The detection of urinary HYAL1 and survivin RNA is a promising noninvasive test for bladder cancer early detection. HYAL1 RNA was more sensitive and specific than urine cytology. Semiquantitative reverse transcriptase-polymerase chain reaction is favored for its high sensitivity and specificity. PMID- 20006859 TI - Magnetic resonance imaging guided prostate biopsy in men with repeat negative biopsies and increased prostate specific antigen. AB - PURPOSE: Undetected cancer in repeat transrectal ultrasound guided prostate biopsies in patients with increased prostate specific antigen greater than 4 ng/ml is a considerable concern. We investigated the tumor detection rate of tumor suspicious regions on multimodal 3 Tesla magnetic resonance imaging and subsequent magnetic resonance imaging guided biopsy in 68 men with repeat negative transrectal ultrasound guided prostate biopsies. We compared results to those in a matched transrectal ultrasound guided prostate biopsy population. Also, we determined the clinical significance of detected tumors. MATERIALS AND METHODS: A total of 71 consecutive patients with prostate specific antigen greater than 4 ng/ml and 2 or greater negative transrectal ultrasound guided prostate biopsy sessions underwent multimodal 3 Tesla magnetic resonance imaging. In 68 patients this was followed by magnetic resonance imaging guided biopsy directed toward tumor suspicious regions. A matched multisession transrectal ultrasound guided prostate biopsy population from our institutional database was used for comparison. The clinical significance of detected tumors was established using accepted criteria, including prostate specific antigen, Gleason grade, stage and tumor volume. RESULTS: The tumor detection rate of multimodal 3 Tesla magnetic resonance imaging guided biopsy was 59% (40 of 68 cases) using a median of 4 cores. The tumor detection rate was significantly higher than that of transrectal ultrasound guided prostate biopsy in all patient subgroups (p <0.01) except in those with prostate specific antigen greater than 20 ng/ml, prostate volume greater than 65 cc and prostate specific antigen density greater than 0.5 ng/ml/cc, in which similar rates were achieved. Of the 40 patients with identified tumors 37 (93%) were considered highly likely to harbor clinically significant disease. CONCLUSIONS: Multimodal magnetic resonance imaging is an effective technique to localize prostate cancer. Magnetic resonance imaging guided biopsy of tumor suspicious regions is an accurate method to detect clinically significant prostate cancer in men with repeat negative biopsies and increased prostate specific antigen. PMID- 20006860 TI - Body mass index as a predictor of urological disease and outcomes-overly simplistic? PMID- 20006861 TI - Absolute preoperative C-reactive protein predicts metastasis and mortality in the first year following potentially curative nephrectomy for clear cell renal cell carcinoma. AB - PURPOSE: C-reactive protein is an inflammatory biomarker associated with tumor burden and metastasis in renal cell carcinoma. Recent studies suggest that preoperative C-reactive protein predicts metastasis and mortality after nephrectomy for localized renal cell carcinoma. However, these studies dichotomized C-reactive protein (typically 10 mg/l or greater vs less than 10 mg/l). Considering the continuous range of C-reactive protein (less than 1 mg/l to greater than 100 mg/l) we assessed the ability of absolute preoperative C reactive protein to predict metastases and mortality as a continuous variable. MATERIALS AND METHODS: Patients with clinically localized (T1-T3N0M0) clear cell renal cell carcinoma were followed for 1 year postoperatively. Metastases were identified radiologically and mortality was determined by death certificate. Univariate and multivariate binary logistic regression analyses examined 1-year relapse-free survival and overall relative survival across patient and disease characteristics. RESULTS: Of the 130 patients in this study metastases developed in 24.6% and 10.8% of the patients died. Mean (SD) preoperative C-reactive protein for patients in whom metastases did and did not develop was 89.17 (74.17) and 9.16 (30.62) mg/l, respectively. Mean preoperative C-reactive protein for patients who did and did not die was 102.61 (77.32) and 19.52 (46.10) mg/l, respectively. On multivariate analysis SSIGN score (p <0.001) and preoperative C reactive protein (B 0.027, SE 0.003, p <0.001) were significant predictors of relapse-free survival, and preoperative platelets (p = 0.009) and preoperative C reactive protein (B 0.011, SE 0.008, p <0.001) were significant predictors of overall relative survival. CONCLUSIONS: Absolute preoperative C-reactive protein is a robust predictor of metastasis and mortality after nephrectomy for localized renal cell carcinoma. Clinicians should consider absolute preoperative C-reactive protein to identify high risk patients for closer surveillance or additional therapy. In addition, predictive algorithms and models of metastasis should consider incorporating C-reactive protein as a continuous variable to maximize predictive ability. PMID- 20006863 TI - Editorial comment. PMID- 20006864 TI - Transvaginal mesh for cystocele repair. PMID- 20006865 TI - Evaluation of and treatment for monosymptomatic enuresis: a standardization document from the International Children's Continence Society. AB - PURPOSE: We provide updated, clinically useful recommendations for treating children with monosymptomatic nocturnal enuresis. MATERIALS AND METHODS: Evidence was gathered from the literature and experience was gathered from the authors with priority given to evidence when present. The draft document was circulated among all members of the International Children's Continence Society as well as other relevant expert associations before completion. RESULTS: Available evidence suggests that children with monosymptomatic nocturnal enuresis could primarily be treated by a primary care physician or an adequately educated nurse. The mainstays of primary evaluation are a proper history and a voiding chart. The mainstays of primary therapy are bladder advice, the enuresis alarm and/or desmopressin. Therapy resistant cases should be handled by a specialist doctor. Among the recommended second line therapies are anticholinergics and in select cases imipramine. CONCLUSIONS: Enuresis in a child older than 5 years is not a trivial condition, and needs proper evaluation and treatment. This requires time but usually does not demand costly or invasive procedures. PMID- 20006866 TI - 7-year oncological outcomes after laparoscopic and open partial nephrectomy. AB - PURPOSE: Open partial nephrectomy has proven long-term oncological efficacy. Long term outcomes of laparoscopic partial nephrectomy are pending. We present the long-term outcomes of patients undergoing laparoscopic or open partial nephrectomy for a single cT1 renal cortical tumor 7 cm or less. MATERIALS AND METHODS: Of 2,246 patients undergoing partial nephrectomy for a single cT1 tumor (1999 to 2008), minimum 7-year followup was available in 77 and 310, and minimum 1-year followup was available in 672 and 944 after laparoscopic and open partial nephrectomy, respectively. Survival and recurrence data obtained from medical records, radiographic reports and patient contact were analyzed retrospectively. RESULTS: Median followup after laparoscopic and open partial nephrectomy was 4.0 and 5.7 years, respectively. Oncological outcomes were excellent in both groups. On multivariable analysis predictors of all cause mortality included advancing age (p <0.0001), comorbidity (p <0.0001) and preoperative renal dysfunction (p = 0.0001) but not tumor size (p = 0.6) or operative approach (laparoscopic vs open partial nephrectomy, p = 0.06). Cancer recurred infrequently and only rarely caused mortality after laparoscopic or open partial nephrectomy. At 7 years metastasis-free survival was 97.5% and 97.3% (p = 0.47) after laparoscopic and open partial nephrectomy, respectively. After accounting for baseline differences between the cohorts using propensity score matching 7-year metastasis-free survival was similar after laparoscopic and open partial nephrectomy. CONCLUSIONS: Laparoscopic and open partial nephrectomy appear to provide similar long-term overall and cancer specific survival in patients undergoing partial nephrectomy for clinical stage T1 (7 cm or less) renal cortical tumors. Oncological outcomes at 7 years after laparoscopic and open partial nephrectomy are excellent with the majority (97%) of patients experiencing metastasis-free survival. PMID- 20006869 TI - Pediatric Urinary Incontinence--shouldn't we speak the same language? PMID- 20006870 TI - Editorial comment. PMID- 20006871 TI - Editorial comment. PMID- 20006872 TI - The prognostic value of erythrocyte polyamine in the post-nephrectomy stratification of renal cell carcinoma specific mortality. AB - PURPOSE: The polyamines spermine and spermidine are ubiquitous polycationic structures which are essential for cell proliferation and differentiation. Circulating polyamines, spermine and spermidine, represent valuable prognostic markers in prostate cancer, acute leukemia and supratentorial malignant glioma. We tested whether spermine and spermidine could improve the prognostic ability of several established predictors of cancer specific mortality after partial or radical nephrectomy for renal cell carcinoma. MATERIALS AND METHODS: Testing was performed on 399 patients with stages T(1-4), N(0-2), M(0-1) renal cell carcinoma who were treated with radical or partial nephrectomy at a single institution between 1990 and 2007. Univariable and multivariable Cox regression models tested the prognostic ability of spermine and spermidine levels in cancer specific mortality predictions. Covariates consisted of TNM stage, Fuhrman grade, tumor size and symptom classification. Harrell's concordance index (c-index) quantified accuracy and 200 bootstrap resamples were used to correct for overfit bias. RESULTS: The 5-year cancer specific mortality-free survival of patients with spermine levels 3 or less, 3.1 to 8, 8.1 to 13 and greater than 13 nmol/8x10(9) erythrocytes was 88.8%, 75.8%, 40.2% and 21.8%, respectively. Similarly the 5 year cancer specific mortality-free survival of patients with spermidine levels 12 or less, 12.1 to 15, 15.1 to 21 and greater than 21 nmol/8x10(9) erythrocytes was 79.0%, 56.6%, 53.2% and 27.4%, respectively. On multivariable analyses addressing cancer specific mortality after surgery spermine (p = 0.007) and spermidine (p = 0.04) achieved independent predictor status. Consideration of spermine and spermidine also improved the accuracy of established cancer specific mortality predictors by 2.2% (p <0.001). CONCLUSIONS: Spermine and spermidine may significantly improve the prognostic value of established cancer specific mortality predictors after partial or radical nephrectomy for all stages of renal cell carcinoma. Independent external validation of our findings is required. PMID- 20006873 TI - Editorial comment. PMID- 20006874 TI - Editorial comment. PMID- 20006875 TI - Unintended consequences of laparoscopic surgery on partial nephrectomy for kidney cancer. AB - PURPOSE: Recent evidence suggests that partial nephrectomy may be associated with improved survival compared to radical nephrectomy for renal cell carcinoma but partial nephrectomy may be underused. We examined whether the introduction of laparoscopic radical nephrectomy contributed to low partial nephrectomy use with time. MATERIALS AND METHODS: We identified all patients treated surgically for renal cell carcinoma in Ontario, Canada between 1995 and 2004 using the Ontario Cancer Registry, a population based tumor registry. A multinomial logistic regression model was used to relate the relative numbers of patients with open and laparoscopic radical nephrectomy, and partial nephrectomy to patient age, gender and surgery year. The partial nephrectomy time trend was investigated by fitting a segmented regression model. RESULTS: Of 7,830 surgically treated patients 7,042 (89.9%) vs 788 (10.1%) underwent radical vs partial nephrectomy. Segmented regression showed a clear change in partial nephrectomy use with time (p = 0.001), such that the odds of partial nephrectomy increased by 18% per year before January 2003 (OR 1.18, 95% CI 1.14-1.23) and subsequently decreased by 12% per year (OR 0.88, 95% CI 0.75-1.02). In the multinomial regression model age and surgery year but not gender were independently associated with partial nephrectomy. CONCLUSIONS: Partial nephrectomy use for renal cell carcinoma remains low, particularly in elderly patients. The introduction of laparoscopic radical nephrectomy coincided with decreased uptake and use of partial nephrectomy for renal cell carcinoma. Although it was hypothesized previously, to our knowledge this is the first study to suggest that the introduction of laparoscopy in renal surgery has negatively impacted partial nephrectomy use. PMID- 20006876 TI - Impact of community wide epidemics of antimicrobial resistance on management of urinary tract infections. PMID- 20006877 TI - Robotic cystectomy: its time has come. PMID- 20006878 TI - An update of the Gleason grading system. AB - PURPOSE: An update is provided of the Gleason grading system, which has evolved significantly since its initial description. MATERIALS AND METHODS: A search was performed using the MEDLINE(R) database and referenced lists of relevant studies to obtain articles concerning changes to the Gleason grading system. RESULTS: Since the introduction of the Gleason grading system more than 40 years ago many aspects of prostate cancer have changed, including prostate specific antigen testing, transrectal ultrasound guided prostate needle biopsy with greater sampling, immunohistochemistry for basal cells that changed the classification of prostate cancer and new prostate cancer variants. The system was updated at a 2005 consensus conference of international experts in urological pathology, under the auspices of the International Society of Urological Pathology. Gleason score 2-4 should rarely if ever be diagnosed on needle biopsy, certain patterns (ie poorly formed glands) originally considered Gleason pattern 3 are now considered Gleason pattern 4 and all cribriform cancer should be graded pattern 4. The grading of variants and subtypes of acinar adenocarcinoma of the prostate, including cancer with vacuoles, foamy gland carcinoma, ductal adenocarcinoma, pseudohyperplastic carcinoma and small cell carcinoma have also been modified. Other recent issues include reporting secondary patterns of lower and higher grades when present to a limited extent, and commenting on tertiary grade patterns which differ depending on whether the specimen is from needle biopsy or radical prostatectomy. Whereas there is little debate on the definition of tertiary pattern on needle biopsy, this issue is controversial in radical prostatectomy specimens. Although tertiary Gleason patterns are typically added to pathology reports, they are routinely omitted in practice since there is no simple way to incorporate them in predictive nomograms/tables, research studies and patient counseling. Thus, a modified radical prostatectomy Gleason scoring system was recently proposed to incorporate tertiary Gleason patterns in an intuitive fashion. For needle biopsy with different cores showing different grades, the current recommendation is to report the grades of each core separately, whereby the highest grade tumor is selected as the grade of the entire case to determine treatment, regardless of the percent involvement. After the 2005 consensus conference several studies confirmed the superiority of the modified Gleason system as well as its impact on urological practice. CONCLUSIONS: It is remarkable that nearly 40 years after its inception the Gleason grading system remains one of the most powerful prognostic factors for prostate cancer. This system has remained timely because of gradual adaptations by urological pathologists to accommodate the changing practice of medicine. PMID- 20006879 TI - Prognostic value of body mass index in Korean patients with renal cell carcinoma. AB - PURPOSE: Whether body mass index is a prognostic factor in patients with renal cell carcinoma continues to be debated. We investigated the association between body mass index, and clinical/pathological features and prognosis in a large cohort of Korean patients with renal cell carcinoma. MATERIALS AND METHODS: The medical records of 1,017 patients with renal cell carcinoma who underwent curative surgery between 1988 and 2006 were reviewed. Mean followup was 76.9 months. We analyzed the association of body mass index at surgery with tumor pathological features, and its associations with cancer specific survival and overall survival were evaluated using the Kaplan-Meier method and Cox regression models. Additional survival analysis was performed in a subgroup of 897 patients with T1-4N0M0 disease. RESULTS: Of the 1,017 patients 363 (35.7%), 526 (51.7%) and 128 (12.6%) had a body mass index of less than 23 (normal), 23 to 27.5 (overweight) and 27.5 or greater (obese) kg/m(2), respectively. Overweight and obese patients had less aggressive tumors, such as less lymph node and/or distant metastases (p = 0.001), low pathological T stage (p = 0.047) and low Fuhrman grade (p = 0.033) vs normal weight patients. In terms of cancer specific survival and overall survival multivariate analysis showed that overweight (p = 0.040 and p = 0.047, respectively) and obese (p = 0.024 and p = 0.010, respectively) patients had good survival rates compared to those with a body mass index in the normal range in the cohort (T1-4NallMall) groups. In addition, overweight (p = 0.022 and p = 0.029, respectively) and obese (p = 0.009 and p = 0.002, respectively) status was significantly associated with cancer specific and overall survival in the T1-4N0M0 groups. CONCLUSIONS: Our findings suggest that overweight and obese Korean patients with renal cell carcinoma have more favorable pathological features and a better prognosis than those with a normal body mass index. PMID- 20006881 TI - Editorial comment. PMID- 20006882 TI - Cost analysis of robotic versus open radical cystectomy for bladder cancer. AB - PURPOSE: Recently robotic approaches to cystectomy have been reported, and while clinical and oncological efficacy continues to be evaluated, potential financial costs have not been clearly evaluated. In this study we present a financial analysis using current cost structures and clinical outcomes for robotic and open cystectomy for bladder cancer. MATERIALS AND METHODS: The financial costs of robotic and open radical cystectomy were categorized into operating room and hospital components, and further divided into fixed and variable costs for each. Fixed operating room costs for open cases involved base cost as well as disposable equipment costs while robotic fixed costs included the amortized machine cost as well as equipment and maintenance. Variable operating room costs were directly related to length of surgery. Variable hospital costs were directly related to transfusion requirement and length of stay. The means of the prior 20 cases of robotic and open cystectomy were used to perform a comparative cost analysis. RESULTS: Mean fixed operating room costs for robotic cases were $1,634 higher than for open cases. Operating room variable costs were also higher by a difference of $570, directly related to increased operating room time. Hospital costs were nearly identical for the fixed component while variable costs were $564 higher for the open approach secondary to higher transfusion costs and longer mean length of stay. Based on these findings robotic cystectomy is associated with an overall higher financial cost of $1,640 (robotic $16,248 vs open $14,608). Cost calculators were constructed based on these fixed and variable costs for each surgical approach to demonstrate the expected total costs based on varying operating room time and length of stay. CONCLUSIONS: Robotic assisted laparoscopic radical cystectomy is associated with a higher financial cost (+$1,640) than the open approach in the perioperative setting. However, this analysis is limited by its single institution design and a multicenter followup study is required to provide a more comprehensive analysis. PMID- 20006883 TI - Accuracy of PCA3 measurement in predicting short-term biopsy progression in an active surveillance program. AB - PURPOSE: PCA3 is a prostate specific noncoding mRNA that is significantly over expressed in prostate cancer tissue. Urinary PCA3 levels have been associated with prostate cancer grade and extent, suggesting a possible role in monitoring patients on active surveillance. We assessed the relationship between PCA3 and prostate biopsy results in men in a surveillance program. MATERIALS AND METHODS: Urine specimens were obtained from 294 men with prostate cancer enrolled in the Johns Hopkins surveillance program. The followup protocol included semiannual free and total prostate specific antigen measurements, digital rectal examination and annual surveillance prostate biopsy. Cox proportional hazards regression was used to evaluate the association between PCA3 results and progression on surveillance biopsy (defined as Gleason pattern 4 or 5, more than 2 positive biopsy cores or more than 50% involvement of any core with cancer). RESULTS: Patients with progression on biopsy (12.9%) had a mean PCA3 score similar to that of those without progression (60.0 vs 50.8, p = 0.131). ROC analysis suggested that PCA3 alone could not be used to identify men with progression on biopsy (AUC 0.589, 95% CI 0.496-0.683, p = 0.076). After adjustment for age and date of diagnosis PCA3 was not significantly associated with progression on biopsy (p = 0.15). CONCLUSIONS: In men with low risk prostate cancer who were carefully selected for surveillance the PCA3 score was not significantly associated with short-term biopsy progression. Further analysis is necessary to assess the usefulness of PCA3 in combination with other biomarkers or in selected subsets of patients undergoing surveillance. PMID- 20006884 TI - Robotic radical cystectomy for bladder cancer: surgical and pathological outcomes in 100 consecutive cases. AB - PURPOSE: Radical cystectomy remains the most effective treatment for patients with localized, invasive bladder cancer and recurrent noninvasive disease. Recently some surgeons have begun to describe outcomes associated with less invasive surgical approaches to this disease such as laparoscopic or robotic assisted techniques. We report our maturing experience with 100 consecutive cases of robotic assisted laparoscopic radical cystectomy with regard to perioperative results, pathological outcomes and surgical complications. MATERIALS AND METHODS: A total of 100 consecutive patients (73 male and 27 female) underwent robotic radical cystectomy and urinary diversion at our institution from January 2006 to January 2009 for clinically localized bladder cancer. Outcome measures evaluated included operative variables, hospital recovery, pathological outcomes and complication rate. RESULTS: Mean age of this cohort was 65.5 years (range 33 to 86). Of the patients 61 underwent ileal conduit diversion, 38 received a neobladder and 1 had no urinary diversion (renal failure). Mean operating room time for all patients was 4.6 hours (median 4.3) and mean surgical blood loss was 271 ml (median 250). On surgical pathology 40% of the cases were pT1 or less disease, 27% were pT2, 13% were pT3/T4 disease and 20% were node positive. Mean number of lymph nodes removed was 19 (range 8 to 40). In no case was there a positive surgical margin. Mean days to flatus were 2.1, bowel movement 2.8 and discharge home 4.9. There were 41 postoperative complications in 36 patients with 8% having a major complication (Clavien grade 3 or higher) and 11% being readmitted within 30 days of surgery. At a mean followup of 21 months 15 patients had disease recurrence and 6 died of disease. CONCLUSIONS: We report a relatively large and maturing experience with robotic radical cystectomy for the treatment of bladder cancer providing acceptable surgical and pathological outcomes. These results support continued efforts to refine the surgical management of high risk bladder cancer. PMID- 20006885 TI - Editorial comment. PMID- 20006886 TI - Editorial comment. PMID- 20006887 TI - Performance of prostate cancer prevention trial risk calculator in a contemporary cohort screened for prostate cancer and diagnosed by extended prostate biopsy. AB - PURPOSE: Statistical models such as the Prostate Cancer Prevention Trial risk calculator have been developed to estimate the cancer risk in an individual and help determine indications for biopsy. We assessed risk calculator performance in a large contemporary cohort of patients sampled by extended biopsy schemes. MATERIALS AND METHODS: The validation cohort comprised 3,482 men who underwent a total of 4,515 prostate biopsies. Calculator performance was evaluated by ROC AUC and calibration plots. A multivariate regression model was fitted to address important predictor variables in the validation data set. Prediction error was calculated as the response variable in another multivariate regression model. RESULTS: Using an average of 13 cores per biopsy prostate cancer was detected in 1,862 patients. The calculator showed an AUC of 0.57 to predict all cancers and 0.60 for high grade cancer. Multivariate analysis of the predictive ability of various clinical factors revealed that race and the number of biopsy cores did not predict overall or high grade cancer at biopsy. Prior negative biopsy, patient age and free prostate specific antigen were significantly associated with prediction error for overall and high grade cancer. Race and family history had a significant association with prediction error only for high grade disease. CONCLUSIONS: To our knowledge our external validation of the Prostate Cancer Prevention Trial risk calculator was done in the largest cohort of men screened for prostate cancer to date. Results suggest that the current calculator remains predictive but does not maintain initial accuracy in contemporary patients sampled by more extensive biopsy schemes. Data suggest that the predictive ability of the calculator in current clinical practice may be improved by modeling contemporary data and/or incorporating additional prognostic variables. PMID- 20006888 TI - Pathological findings and prostate specific antigen outcomes after radical prostatectomy in men eligible for active surveillance--does the risk of misclassification vary according to biopsy criteria? AB - PURPOSE: We compared the pathological findings and prostate specific antigen outcome after radical prostatectomy in men eligible for active surveillance according to 3 biopsy inclusion criteria. MATERIALS AND METHODS: The study population included 177 men eligible for active surveillance who fulfilled clinicobiological criteria and biopsy criteria as group 1-less than 3 positive cores and less than 3 mm total tumor length, group 2-less than 3 positive cores with cancer involvement of less than 50% in any core and group 3-less than 33% of positive cores. Prostate specific antigen density cutoffs were also studied in these groups. Pathological findings on radical prostatectomy specimens and biochemical recurrence-free survival were studied. Median followup after radical prostatectomy was 34 months. RESULTS: A majority of Gleason score 6 disease was observed in group 1 (51.7%) whereas a majority of Gleason score 7 or greater disease was reported in groups 2 (53.6%) and 3 (55.4%). Extracapsular extension was noted in 17.5% of radical prostatectomy specimens in group 3 vs 11.2% in group 1 (p = 0.175). The risk of overall unfavorable disease (defined as pT3-4 stage and/or Gleason score 8 or greater) was significantly higher in men with cancer involvement of 3 mm or greater on initial biopsy (27.3% vs 13.5%, respectively, p = 0.023). The 3-year biochemical recurrence-free survival rate was 94.0% and was not affected by the 3 active surveillance definitions. CONCLUSIONS: Even with the use of a 21-core biopsy protocol the rate of unfavorable disease in radical prostatectomy specimens remains increased in men eligible for active surveillance. Patients must be informed of this risk of misclassification which ranges from 20% to 28% in men who fulfill the less stringent biopsy criteria. PMID- 20006889 TI - Editorial comment. PMID- 20006891 TI - Use of haloperidol and azaperone for stress control in roe deer (Capreolus capreolus) captured by means of drive-nets. AB - The physical capture of wild ungulates is performed for different purposes when anaesthesia in field conditions is not possible or advisable. The use of tranquilizers may contribute to improved welfare of captured animals. We studied the effect of haloperidol and azaperone on the stress response of roe deer (Capreolus capreolus) through the study of physiological, haematological and serum biochemical parameters. Thirty one roe deer were drive-net captured and randomly injected with haloperidol (0.30+/-0.04 mg/kg IM; n=13), azaperone (0.43+/-0.07 mg/kg IM; n=11) or saline (0.5 mL IM; n=7), and restrained for 3h. The interindividual variability of heart rate was lower in the treated deer, suggesting a calming effect, and erythrocyte and biochemical parameters indicated vasodilation, splenic sequestration, hemodilution, improvement of renal perfusion and a protective effect on muscle. These results support the suitability of using either azaperone or haloperidol in capture operations of roe deer, in order to reduce stress and prevent its adverse effects. PMID- 20006892 TI - HBsAg-vectored vaccines simultaneously deliver CTL responses to protective epitopes from multiple viral pathogens. AB - We have previously demonstrated that the potent immunogenicity of hepatitis B surface antigen (HBsAg) may be exploited to deliver foreign antigens for cytotoxic T-lymphocyte (CTL) induction. Here we demonstrate that a single low dose immunization with rHBsAg DNA is sufficient to prime for CTL responses against encoded foreign epitope and that the responses may be recalled many months after immunization. We show that simultaneous disease-protective CTL responses restricted through a diversity of MHC class I haplotypes are elicited by recombinant (r) HBsAg DNA containing multiple viral epitopes appended as a C' terminal polyepitope or encoded individually within the HBsAg polypeptide. CTL responses delivered by rHBsAg DNA were elicited in the presence of HBsAg-directed antibody. These studies vindicate the use of HBsAg as a powerful vector to deliver CTL responses to foreign antigen and have implications for a multidisease vaccine applicable to an MHC-polymorphic population. PMID- 20006893 TI - Photodestruction of BDE-99 in micellar solutions of nonionic surfactants of Brij 35 and Brij 58. AB - Currently, soil contamination by polybrominated diphenyl ethers (PBDEs) at e waste recycling sites in China is of significant environmental concern and requires the identification of appropriate remediation technologies. In this paper, photodestruction of a model PBDE congener (BDE-99) in selected UV/surfactant systems was investigated, using a RPR-200 Rayonet photochemical reactor equipped with two low-pressure mercury lamps. BDE-99 photodegradation in the micellar solutions of nonionic surfactants of Brij 35 and Brij 58 all followed pseudo first-order kinetics and the photodegradation quantum yields were 1.8-2.4 times as high as that in water alone. The photodegradation was comparably enhanced by Brij 35 and Brij 58, and the quenching effect of dissolved oxygen (DO) was eliminated in their presence. Mono- to tetra-BDEs were identified as one group of BDE-99 photoproducts, indicating that one of BDE-99 photodegradation pathways was through a sequential loss of Br. The pattern of PBDE photoproducts showed the photodebromination of BDE-99 readily occurred on the more heavily substituted phenyl ring. Mono- to tetra-polybrominated dibenzofurans (PBDFs) were identified as another group of photoproducts, indicating BDE-99 can also photodegrade through a dibenzofuran-type ring closure process via an intramolecular elimination of HBr. In addition, the formation of these more toxic photoproducts (PBDFs) was not related to the presence of DO. Finally, the later disappearance of mono-BDEs and mono-BDFs suggested the UV exposure time would be an important technical parameter if the technology of UV photodestruction of PBDEs in surfactant micellar solutions is applied to PBDE-contaminated soil remediation. PMID- 20006894 TI - Negative effects of fluoranthene on the ecophysiology of tomato plants (Lycopersicon esculentum Mill) Fluoranthene mists negatively affected tomato plants. AB - Cherry tomato plants (Lycopersicon esculentum Mill) were sprayed with fluoranthene and mixture of fluoranthene and mannitol solutions for 30d. The exposure was carried out in growth chambers in field conditions, and the air was filtered through charcoal filters to remove atmospheric contaminants. Plants were sprayed with 10microM fluoranthene as mist until they reached the fruiting stage, and the eco-physiological parameters were measured to determine the effects of the treatments. We measured CO(2) uptake and water vapour exchange, chlorophyll fluorescence, leaf pigment contents, visual symptoms and biomass allocation. Fluoranthene which was deposited as mist onto leaves negatively affected both growth and the quality of tomato plants, while other treatments did not. The photosynthetic rate measured at saturated irradiance was approximately 37% lower in fluoranthene-treated plants compared with the control group. Other variables, such as stomata conductance, the photochemical efficiency of PSII in the dark, Chl a, Chl b, and the total chlorophyll contents of the tomato leaves were significantly reduced in the fluoranthene-treated plants. Tomato plants treated with fluoranthene showed severe visible injury symptoms on the foliage during the exposure period. Mannitol (a reactive oxygen scavenger) mitigated effects of fluoranthene; thus, reactive oxygen species generated through fluoranthene may be responsible for the damaged tomato plants. It is possible for fluoranthene to decrease the aesthetic and hence the economic value of this valuable crop plant. PMID- 20006895 TI - Changes in brain monoamine levels in neonatal rats exposed to bisphenol A at low doses. AB - To examine whether exposure to bisphenol A (BPA) at low levels affect brain function, monoamine concentrations in hippocampus, striatum and brain stem, were investigated in neonatal male rats injected intracranially with BPA at 0 10microgkg(-1). Significant increases of serotonin (5-HT) in hippocampus, 5-HIAA and 5-HIAA/5-HT in brain stem, dopamine (DA) and DOPAC in striatum were observed at 28d after the injection on postnatal day 2. At 7d after the injection, increases in 5-HT and norepinephrine (NE) and decreases in DOPAC and 5-HIAA were observed in hippocampus. To investigate the degradation of BPA in brain, we also measured BPA concentrations of whole neonatal rat brain. Free BPA disappeared from brain tissues within 5h, even when the highest dose (1000microgkg(-1)) was injected. The present results suggest that BPA exposure at lower doses than environmentally relevant levels may have a great impact on monoamine levels in neonatal brain over 28d after its disappearance. PMID- 20006896 TI - Distribution of phthalate esters in alluvial sediment: a case study at JiangHan Plain, Central China. AB - This study aimed to investigate the sources and distribution of phthalate esters (PAEs) in alluvial sediment of humid climate regions where water cycle is very active in order to reveal their behavior of transport from topsoil and/or surface water to deep sediment and groundwater. Topsoil and deeper sediment samples were collected from nine and seventeen sites in July 2007, and January 2008, respectively, from the eastern part of JiangHan Plain, Central China. On each site, samples were collected at every 20-40cm depth within the sediment profiles. Contents of 16 PAEs were detected for each sample. summation operator(16)PAEs contents in the topsoil ranged from 252.6 to 2515.7ngg(-1), with an average value of 926.8ngg(-1). Di(2-ethylhexyl) phthalate (DEHP), di-isobutyl phthalate (DiBP), di-n-butyl phthalate (DnBP), and diethyl phthalate (DEP) were the dominant PAE species. The horizontal distribution of PAEs was related to time (season), cultivation type, distance and exact position from surface water. DEP existed only on the upper layer of soil due to its rapid degradation. However, DEHP, DnBP, and DiBP could be transported downward into deep sediment even though large amount of them were lost due to biodegradation and adsorption. On the other hand, DEHP, DnBP, and DiBP could be transported into deep sediment along with the horizontal flow of shallow groundwater from surface water, such as Yangtze River, Hanjiang River, and Honghu Lake. PMID- 20006897 TI - Comment on Urban et al. "Assessment of human health risks posed by consumption of fish from the Lower Passaic River (LPR), New Jersey" (2009, doi:10.1016/jscitotenv.2009.03.004). AB - Urban et al. (2009) presented a human health risk assessment for the Lower Passaic River that is not consistent with the United States Environmental Protection Agency (USEPA) risk assessment guidance for Superfund cited in the article, because it is based on a fish ingestion rate that underestimates angler exposure to contaminated fish, does not evaluate exposure to contaminated crab, and underestimates the cancer risks and non-cancer health hazards associated with these exposure pathways. USEPA recommends that people follow the health advisories for the Lower Passaic River, available online at http://www.state.nj.us/dep/dsr/2009FishAdvisoryBrochure.pdf. PMID- 20006898 TI - Heavy metal stabilization in contaminated road-derived sediments. AB - There is increasing interest in the stabilization of heavy metals in road-derived sediments (RDS), to enable environmentally responsible reuse applications and circumvent the need for costly landfill disposal. To reduce the mobility of heavy metals (i.e. Cu, Pb and Zn) the effectiveness of amendments using phosphate, compost and fly ash addition were investigated using batch leaching experiments. In general, phosphate amendments of RDS were found to be ineffective at stabilizing heavy metals, despite being used successfully in soils. Phosphate amendment resulted in enhanced concentrations of dissolved organic carbon (DOC), which increased the solubilisation of heavy metals via complexation. Amendment with humified organic matter (compost) successfully stabilized Cu and Pb in high DOC leaching RDS with an optimum loading of 15-20% (w/w). Compost, however, was ineffective at stabilizing Zn. Increasing the pH by amending RDS/compost blends with 2.5-15% (w/w) coal fly ash resulted in the stabilization of Zn, Cu and Pb. However, above a pH of approximately 7.5 and 8 enhanced leaching of organic matter resulted in an increase in leached Cu and Pb, respectively. Accordingly, the optimum level of fly ash amendment for the RDS/compost blends was estimated to be ca. 10%. Boosted regression trees analysis (BRT) of the data revealed that DOC accounted for 56% and 65% of the Cu and Pb leaching, respectively, whereas pH only accounted for ca. 18% of Cu and Pb leaching. RDS sample characteristics (i.e. metal concentrations, size fractionation and organic matter content) were more important at reconciling the leaching concentrations of copper Cu (27%) than Pb (16%). The most important parameter explaining Zn leaching was pH. Overall, the choice of a suitable stabilization agent/s depends on the composition of RDS with respect to the amount of organic matter present, and the sorption chemistry of the heavy metal of interest. PMID- 20006899 TI - Lupus Anticoagulant-Hypoprothrombinemia syndrome (HLAS): report of one case in a familial infectious context. PMID- 20006900 TI - Ovarian cancer tumor infiltrating T-regulatory (T(reg)) cells are associated with a metastatic phenotype. AB - OBJECTIVE: The objective of this study was to examine the clinicopathologic correlates of T-regulatory (T(reg)) cell infiltration in serous ovarian cancers and to define gene signatures associated with high T(reg)s. METHODS: Tumor infiltrating T(reg) and cytotoxic T-cells (CTLs) were quantitated in 232 primary serous ovarian cancers by immunostaining for FOXP3 and CD8. Expression microarray analysis was performed in a subset of 48 advanced cancers with the highest and lowest numbers of infiltrating T(reg)s and a genomic signature was developed using binary regression. ANOVA analysis was performed to assess the most differentially expressed genes and these genes were further assessed using Ingenuity Pathway Analysis (IPA) software. RESULTS: High T(reg) infiltration in ovarian cancers was associated with high grade (p<0.0001), advanced stage (p=0.004) and suboptimal debulking (p<0.04), but not with survival. In contrast, high tumor infiltrating CD8 CTL infiltration was associated with favorable survival (median survival 48.7 vs. 34.6 months, p=0.01). A microarray-based genomic signature for high tumor-infiltrating T(reg) cells had a 77% predictive accuracy using leave-one-out cross-validation. ANOVA of microarray data revealed the antigen presentation pathway as the most differentially expressed canonical pathway (p<0.00001) between cancers with high and low T(reg) cells. CONCLUSIONS: These data suggest that there may be an association between increased T(reg) cell infiltration in ovarian cancers and advanced stage. Increased T(reg) infiltration is characterized by a genomic signature enriched with several immunologic pathway genes. Therapeutic strategies that reduce tumor infiltrating T(reg) cells are under investigation and may prove useful in ovarian cancers with high numbers of these cells. PMID- 20006901 TI - Shotgun redox proteomics in sub-proteomes trapped on functionalised beads: Identification of proteins targeted by oxidative stress. AB - If reactive oxygen species (ROS) levels exceed antioxidant defences, oxidative stress occurs; a common response to environmental pollutants. Proteins absorb ~70% of ROS, altering amino acid side-chains. Cys (-SH) oxidises to sulphenic ( SOH), sulphinic (-SO(2)H), cysteic (-SO(3)H) acids and disulphide bridges (-S-S ). Two-dimensional electrophoresis (2DE) under-selects certain protein categories (e.g. extreme pI, small proteins) so activated thiol sepharose (ATS) was used to select sub-proteomes of thiol-containing proteins in menadione-exposed Escherichia coli. ATS bound thiol-containing proteins (but not oxidised thiols) via mixed disulphides. Tryptic digestion of bead-bound proteins was followed by LC-tandem MS. Many proteins were identified in controls with significantly fewer in menadione-treated cells (e.g. chaperonins, transcription/translation-related and ribosomal proteins; aminoacyl tRNA synthetases and metabolic enzymes. Non denaturing ATS capture (followed by reduction) demonstrated lower specific activities of key enzymes which is attributed to thiol oxidation. This method may be generally useful in ecotoxicology for identification of oxidative stress targets. PMID- 20006902 TI - Man induced change in community control in the north-western Black Sea: The top down bottom-up balance. AB - The present study shows how marine commercial fish food webs dramatically changed in the north-western Black Sea on both pelagic and benthic environments. Fisheries landings, diversity and equitability strongly decreased between 1965 1970 and 2001-2005. Fishes adapted their feeding behaviour to the increasingly low species diversity of the Black Sea communities. Their food web became poor and simplified following the loss of many top predator species and their trophic links. Linkage density, connectivity and Lyapunov stability proxy strongly decreased. The north-western Black Sea system switched from a complex top-down and bottom-up functioning pattern to a dominantly bottom-up functioning pattern. This study contributes to a better understanding of these transformations within the Danube-Black Sea system in the last decades. An attempt is made to relate these changes with river inputs, fisheries and coastal pollution. PMID- 20006903 TI - Seasonal variations and alterations of sex steroid levels during the reproductive cycle of male roach (Rutilus rutilus). AB - Seasonal variations of plasma steroid concentrations i.e. progesterone (P), 11 ketotestosterone (11-KT) and 17-beta-estradiol (E2) were determined immunoenzymatically during a whole reproductive cycle in male roach (Rutilus rutilus) caught in a reference site. Plasma 11-KT concentrations were significantly correlated with gonad growth, expressed as the gonado-somatic index (R2 =0.60; p<0.05) and highest concentrations (757 +/- 99 pg ml-1 ) coincided with the final testis maturation in March. E2 and P concentrations peaked once during the reproductive cycle. E2 synthesis was significantly induced during the spawning period while P concentration peaked at the very start of the gametogenesis (June) thus suggesting specific roles of these steroids in the reproductive cycle. Comparison of reference levels were then made with plasma steroid concentrations from male roach sampled in polluted areas in the North of France. A significant decrease of E2 (50-60%) and 11-KT (76-84%) was measured, indicating that endocrine disrupting compounds may have interfered with the normal sex steroid synthesis. Contrary to the E2 and 11-KT sex steroids, plasma P concentration was not significantly affected in fish inhabiting impacted areas. PMID- 20006905 TI - Hypopyon in patients with uveitis. AB - PURPOSE: To evaluate the risk of and risk factors for hypopyon among patients with uveitis and to evaluate the risk of visual changes and complications after hypopyon. DESIGN: Retrospective cohort study. PARTICIPANTS: Patients with uveitis at 4 academic ocular inflammation subspecialty practices. METHODS: Data were ascertained by standardized chart review. MAIN OUTCOME MEASURES: Prevalence and incidence of hypopyon, risk factors for hypopyon, and incidence of visual acuity changes and ocular complications after hypopyon. RESULTS: Among 4911 patients with uveitis, 41 (8.3/1000) cases of hypopyon were identified at the time of cohort entry. Of these, 2885 initially free of hypopyon were followed over 9451 person-years, during which 81 patients (2.8%) developed hypopyon (8.57/1000 person-years). Risk factors for incident hypopyon included Behcet's disease (adjusted relative risk [RR]=5.30; 95% confidence interval [CI], 2.76-10.2), spondyloarthropathy (adjusted RR=2.86; 95% CI, 1.48-5.52), and human leukocyte antigen (HLA)-B27 positivity (adjusted RR=2.04; 95% CI, 1.17-3.56). Patients with both a spondyloarthropathy and HLA-B27 had a higher risk than either factor alone (crude RR=4.39; 95% CI, 2.26-8.51). Diagnosis of intermediate uveitis (+/- anterior uveitis) was associated with a lower risk of hypopyon (with respect to anterior uveitis only, adjusted RR=0.35; 95% CI, 0.15-0.85). Hypopyon incidence tended to be lower among patients with sarcoidosis (crude RR=0.22; 95% CI, 0.06 0.90; adjusted RR=-0.28; 95% CI, 0.07-1.15). Post-hypopyon eyes and eyes not developing hypopyon had a similar incidence of band keratopathy, posterior synechiae, ocular hypertension, hypotony, macular edema, epiretinal membrane, cataract surgery, or glaucoma surgery. Post-hypopyon eyes were more likely than eyes not developing hypopyon to gain 3 lines of vision (crude RR=1.54; 95% CI, 1.05-2.24) and were less likely to develop 20/200 or worse visual acuity (crude RR=0.41; 95% CI, 0.17-0.99); otherwise, visual outcomes were similar in these groups. CONCLUSIONS: Hypopyon is an uncommon occurrence in patients with uveitis. Risk factors included Behcet's disease, HLA-B27 positivity, and spondyloarthropathy. Intermediate uveitis cases (+/- anterior uveitis) had a lower risk of hypopyon. On average, post-hypopyon eyes were no more likely than other eyes with uveitis to develop structural ocular complications or lose visual acuity. PMID- 20006904 TI - Genome-wide census and expression profiling of chicken neuropeptide and prohormone convertase genes. AB - Neuropeptides regulate cell-cell signaling and influence many biological processes in vertebrates, including development, growth, and reproduction. The complex processing of neuropeptides from prohormone proteins by prohormone convertases, combined with the evolutionary distance between the chicken and mammalian species that have experienced extensive neuropeptide research, has led to the empirical confirmation of only 18 chicken prohormone proteins. To expand our knowledge of the neuropeptide and prohormone convertase gene complement, we performed an exhaustive survey of the chicken genomic, EST, and proteomic databases using a list of 95 neuropeptide and 7 prohormone convertase genes known in other species. Analysis of the EST resources and 22 microarray studies offered a comprehensive portrait of gene expression across multiple conditions. Five neuropeptide genes (apelin, cocaine-and amphetamine-regulated transcript protein, insulin-like 5, neuropeptide S, and neuropeptide B) previously unknown in chicken were identified and 62 genes were confirmed. Although most neuropeptide gene families known in human are present in chicken, there are several gene not present in the chicken. Conversely, several chicken neuropeptide genes are absent from mammalian species, including C-RF amide, c-type natriuretic peptide 1 precursor, and renal natriuretic peptide. The prohormone convertases, with one exception, were found in the chicken genome. Bioinformatic models used to predict prohormone cleavages confirm that the processing of prohormone proteins into neuropeptides is similar between species. Neuropeptide genes are most frequently expressed in the brain and head, followed by the ovary and small intestine. Microarray analyses revealed that the expression of adrenomedullin, chromogranin A, augurin, neuromedin-U, platelet-derived growth factor A and D, proenkephalin, relaxin-3, prepronociceptin, and insulin-like growth factor I was most susceptible (P-value<0.005) to changes in developmental stage, gender, and genetic line among other conditions studied. Our complete survey and characterization facilitates understanding of neuropeptides genes in the chicken, an animal of importance to biomedical and agricultural research. PMID- 20006906 TI - Does the presence of an epiretinal membrane alter the cleavage plane during internal limiting membrane peeling? AB - PURPOSE: To determine whether the presence of a clinically and/or microscopically detectable epiretinal membrane (ERM) alters the cleavage plane during internal limiting membrane (ILM) peeling. DESIGN: Retrospective, observational, immunohistochemical study of ILM specimens using archival formalin-fixed, paraffin-embedded tissue. PARTICIPANTS: Fifty-one patients who had had ILM excision. METHODS: Fifty-one ILM specimens peeled during vitrectomy for various etiologies were examined by light microscopy. The removal of ILM was assisted using Trypan blue (n = 30), indocyanine green (n = 7), or brilliant blue G (n = 14). Monoclonal antibodies to glial fibrillary acidic protein and to neurofilament protein were used to detect glial or neuronal cells respectively on the vitreous or retinal surfaces of the ILM. Specimens were divided into 2 groups: ILM peeled for full-thickness macular hole (MH; n = 31) and ILM peeled after removal of clinically detectable ERM (n = 20). MAIN OUTCOME MEASURES: Primary outcome measure was the localization of immunohistochemical markers to neuronal or glial cells on the vitreous or retinal surfaces of ILM. The secondary outcome measure was the correlation of the results of the primary measure with the dyes used to facilitate ILM peeling. RESULTS: Glial and/or neuronal cells were detected on the retinal surface of the ILM in 10 of 31 (32%) of the MH ILM specimens and in 13 of 20 (65%) of the ILM peeled after ERM excision; the difference was significant (P = 0.02). There was no association between the presence of neuronal and glial cells with the type of dye used (P = 0.2). Of the 23 ILM specimens with cells attached to the retinal surface, 21 (91%) were associated with clinical and/or histologic evidence of ERM and 2 (9%) were not. The correlation between the presence of cells on the vitreous and the retinal surfaces of ILM was high (P<0.0001). CONCLUSIONS: The findings suggest that ERM may be associated with sub-ILM changes that alter the plane of separation during ILM peeling. This study does not confirm any influence of dyes on the cleavage plane during surgery. PMID- 20006907 TI - Diagnostic ability of Heidelberg Retina Tomography in detecting glaucoma in a population setting: the Singapore Malay Eye Study. AB - OBJECTIVE: To evaluate the performance of the Heidelberg Retina Tomograph II (HRT II, Heidelberg Engineering, Heidelberg, Germany) in diagnosing glaucoma in a population setting. DESIGN: Population-based cross-sectional study. PARTICIPANTS: Of 3280 Malay persons aged 40 to 80 years who participated in the survey, 112 subjects (124 eyes) with glaucoma and a subset of 196 bilaterally normal subjects (392 eyes) were included for the evaluation of diagnostic ability of HRT II. METHODS: All glaucomatous and normal eyes underwent standardized ophthalmic assessment (including automated perimetry) and HRT II. Glaucoma was defined according to International Society for Geographical and Epidemiological Ophthalmology criteria. Area under the curve (AUC) receiver operating characteristic (ROC), sensitivity, and specificity were used to evaluate the diagnostic performance of HRT II algorithms. Marginal logistic regression models were used to evaluate the influence of optic disc size on the results of HRT II algorithms. MAIN OUTCOME MEASURES: The HRT II algorithms: linear discriminant functions (LDFs) by Mikelberg et al (LDF1), Burk et al (LDF2), Bathija et al (LDF3), and Moorfields regression analysis (MRA). The MRA was subdivided into MRA1 with "borderline" outcomes as positive and MRA2 with "borderline" outcomes as negative. RESULTS: Subjects with cataract, visual impairment, astigmatism, and greater negative spherical equivalent, and of older age were more likely to yield lower quality images. For analyses by eye, AUCs were 0.789, 0.704, 0.755, 0.754, and 0.762 for MRA1, MRA2, LDF1, LDF2, and LDF3, respectively. At 85% specificity, sensitivities were 62.1%, 65.3%, and 66.9% for LDF1, LDF2, and LDF3, respectively. At 95% specificity, these figures decreased to 31.5%, 42.7%, and 45.2%, respectively. The sensitivity and specificity were 71.0% and 86.7% for MRA1 and 43.6% and 97.2% for MRA2, respectively. Similar estimates were found for analyses by person. Larger optic disc size was associated with increased sensitivity and false-positive rate for MRA1, LDF1, and LDF2. LDF1 and LDF3 were least affected by optic disc area, but the sensitivities were moderate and the false-positive rates were high across different optic disc areas. CONCLUSIONS: The current HRT II algorithms are of limited value for population-based glaucoma screening in the Malay population and do not account adequately for optic disc size. PMID- 20006908 TI - Randomized evaluation of spectacles plus alternate-day occlusion to treat amblyopia. AB - PURPOSE: To compare spectacles plus patching >or=8 hours daily 6 days a week with spectacles plus patching >or=8 hours on alternate days to treat amblyopia in children 4 to 5 years of age. DESIGN: Prospective, randomized clinical trial. PARTICIPANTS: Forty children (median age, 4.3 years) with untreated amblyopia and a median best-corrected visual acuity (BCVA) in the amblyopic eye of 0.9 (range, 0.3-1.5) logarithm of the minimum angle of resolution. METHODS: Refractive correction was provided, and the children were randomized to patching >or=8 hours daily 6 days a week or patching >or=8 hours on alternate days. The BCVA, binocular function, and refractive errors were measured repeatedly during the study. MAIN OUTCOME MEASURE: Median change in BCVA of the amblyopic eye after 1 year. RESULTS: The median change in BCVA of the amblyopic eye did not differ significantly between the 2 groups (0.6 log units for daily occlusion; 0.8 log unit for alternate-day occlusion). The final median BCVA in the amblyopic eyes was 0.1 logarithm of the minimum angle of resolution in both groups. Binocular function improved in both groups with no significant differences between the groups at 1 year. The median spherical equivalent refractive error did not change significantly during the study period in the amblyopic eyes in either group; however, a significant increase was found in the fellow eyes in both groups (daily occlusion, P<0.05; alternate-day occlusion, P<0.001). CONCLUSIONS: The magnitude of change in the BCVA 1 year after spectacles plus prescribed alternate day patching was not significantly different than that after spectacles plus prescribed daily patching to treat amblyopia in children 4 to 5 years old. The effect of patching was not separate from that of optical correction with a period of refractive adaptation. Thus, the improvement in visual acuity is a combined effect of spectacle wear and occlusion therapy. PMID- 20006909 TI - Infectious keratitis in 204 586 LASIK procedures. AB - PURPOSE: To investigate the incidence, culture results, risk factors, and visual outcomes of infectious keratitis after LASIK, and examine treatment strategies. DESIGN: Retrospective study. PARTICIPANTS: We included 107 613 patients who underwent LASIK at Clinica Baviera (Instituto Oftalmologico Europeo, Spain) from September 2002 to May 2008. METHODS: The medical records of post-LASIK patients (204 586 eyes) were reviewed to identify cases of infectious keratitis. Incidence, risk factors, clinical course, days to diagnosis, medical and surgical treatment, and final visual outcomes were recorded. MAIN OUTCOME MEASURES: Incidence of post-LASIK infectious keratitis, culture results, response to treatment, and visual outcome. RESULTS: Post-LASIK infectious keratitis was diagnosed in 72 eyes from 63 patients. Onset of infection was early (within 7 days after surgery) in 62.5% of cases. Cultures were positive in 21 of 54 cases in which samples were taken. The most frequently isolated microorganism was Staphylococcus epidermidis (9 cases). Immediate flap lifting and irrigation with antibiotics was performed in 54 eyes; late flap lifting was subsequently required in 10 out of 18 cases initially treated with topical antibiotics alone. One case required flap amputation owing to flap necrosis. Final best spectacle-corrected visual acuity (BSCVA) was >or=20/20 in 38 cases (52.7%) and >or=20/40 in 67 cases (93.05%); final BSCVA was <20/40 in 5 cases (6.94%). CONCLUSIONS: The incidence of post-LASIK infectious keratitis was 0.035% per procedure. Infectious keratitis after LASIK is a potentially vision-threatening complication. The appearance of infections in asymptomatic patients highlights the need for a proper schedule of follow-up appointments. Prompt and aggressive management of this LASIK complication with early flap lifting, scraping, culture, and irrigation with antibiotics is strongly recommended. Proper management can result in preserving useful vision. FINANCIAL DISCLOSURE(S): The authors have no proprietary or commercial interest in any of the materials discussed in this article. PMID- 20006910 TI - Additive effects of combination treatment with anti-inflammatory and neuroprotective agents in experimental autoimmune encephalomyelitis. AB - We studied the effects of combination treatment with an anti-inflammatory agent, interferon (IFN)-beta, and a putative neuroprotective agent, an estrogen receptor (ER)-beta ligand, during EAE. Combination treatment significantly attenuated EAE disease severity, preserved axonal densities in spinal cord, and reduced CNS inflammation. Combining ERbeta treatment with IFNbeta reduced IL-17, while it abrogated IFNbeta-mediated increases in Th1 and Th2 cytokines from splenocytes. Additionally, combination treatment reduced VLA-4 expression on CD4+ T cells, while it abrogated IFNbeta-mediated decreases in MMP-9. Our data demonstrate that combination treatments can result in complex effects that could not have been predicted based on monotherapy data alone. PMID- 20006911 TI - Survival of Campylobacter spp. in poultry meat preparations subjected to freezing, refrigeration, minor salt concentration, and heat treatment. AB - The survival of Campylobacter spp. under defined conditions of freezing (-22 degrees C) was studied in naturally contaminated chicken skin and minced chicken meat. A decline of approximately one log(10) cfu/g was observed after 1 day of freezing. No further significant reduction was achieved by prolonged storage in the freezer, although a tendency for further gradual reduction of the numbers of Campylobacter spp. present was noted. Campylobacter spp. could still be detected qualitatively (per 0.1g) after 84 days. In a second part of this study, the survival of Campylobacter spp. in a typical minced meat preparation (minced meat supplemented with 1.5% salt (NaCl)) stored at refrigeration (4 degrees C) or frozen (-22 degrees C) was studied. No significant reduction of the pathogen was observed if the minced chicken meat was kept at 4 degrees C for 14 days, opposite to approximately one log(10) cfu/g reduction after 1 day when the minced meat preparation was stored in the freezer (-22 degrees C) for 14 days. The latter reduction is imputed to the effect of freezing as mentioned above and not due to the supplementation of NaCl to minced meat or the combination of NaCl and freezing, because similar reductions of Campylobacter spp. were noticed when minced meat (without addition of NaCl) was frozen. Finally, in a third part of the study, the survival of Campylobacter spp. subjected to a heat treatment, conform to consumer-based pan-frying, in inoculated (4.5+/-0.2 cfu/g) as well as naturally contaminated chicken burgers (2.1+/-0.1 cfu/g) was studied. The Campylobacter spp. numbers declined after 2 min (internal temperature reached circa 38 degrees C), where after 4 min (internal temperature reached circa 57.5 degrees C) they dropped below detectable levels (<10 cfu/g). PMID- 20006912 TI - Promoting Excellence in Nursing Education (PENE): Pross evaluation model. AB - The purpose of this article is to examine the Promoting Excellence in Nursing Education (PENE) Pross evaluation model. A conceptual evaluation model, such as the one described here, may be useful to nurse academicians in the ongoing evaluation of educational programs, especially those with goals of excellence. Frameworks for evaluating nursing programs are necessary because they offer a way to systematically assess the educational effectiveness of complex nursing programs. This article describes the conceptual framework and its tenets of excellence. PMID- 20006913 TI - Evaluation of formal methods in hip joint center assessment: an in vitro analysis. AB - BACKGROUND: The hip joint center is a fundamental landmark in the identification of lower limb mechanical axis; errors in its location lead to substantial inaccuracies both in joint reconstruction and in gait analysis. Actually in Computer Aided Surgery functional non-invasive procedures have been tested in identifying this landmark, but an anatomical validation is scarcely discussed. METHODS: A navigation system was used to acquire data on eight cadaveric hips. Pivoting functional maneuver and hip joint anatomy were analyzed. Two functional methods - both with and without using the pelvic tracker - were evaluated: specifically a sphere fit method and a transformation techniques. The positions of the estimated centers with respect to the anatomical center of the femoral head, the influence of this deviation on the kinematic assessment and on the identification of femoral mechanical axis were analyzed. FINDINGS: We found that the implemented transformation technique was the most reliable estimation of hip joint center, introducing a - Mean (SD) - difference of 1.6 (2.7) mm from the anatomical center with the pelvic tracker, whereas sphere fit method without it demonstrated the lowest accuracy with 25.2 (18.9) mm of deviation. Otherwise both the methods reported similar accuracy (<3mm of deviation). INTERPRETATION: The functional estimations resulted in the best case to be in an average of less than 2mm from the anatomical center, which corresponds to angular deviations of the femoral mechanical axis smaller than 1.7 (1.3) degrees and negligible errors in kinematic assessment of angular displacements. PMID- 20006914 TI - Repetitive transcranial magnetic stimulation (rTMS): a possible novel therapeutic approach to dementia with Lewy bodies. AB - Dementia with Lewy bodies (DLB) is characterized clinically by widespread cognitive loss, visual hallucinations, depression, anxiety and extrapyramidal signs (EPS). DLB is sensitive to typical neuroleptics. Repetitive transcranial magnetic stimulation (rTMS) has been studied as a potential treatment in many neurological disorders, which has been proved to have positive effect on a variety of cognitive functions, hallucinations of schizophrenia, major depression, anxiety, the Parkinson's disease. This report proposes that rTMS may represent an alternative strategy for the treatment of dementia with Lewy bodies. PMID- 20006915 TI - Max Delbruck, Lafcadio Hearn and The Tale of the Heike. PMID- 20006916 TI - Nitrous oxide (laughing gas) inhalation as an alternative to electroconvulsive therapy. AB - Electroconvulsive therapy (ECT) is used widely in the treatment of psychiatric conditions; however, its use is not without controversy with some recommending a moratorium on its clinical use. Complications and side effects of ECT include memory loss, injury, problems originating from sympathetic stimulation such as arrhythmias and myocardial ischemia and the risk of general anesthesia. Nitrous oxide (laughing gas) could potentially substitute for ECT as it shares some similar effects, has potential beneficial properties for these psychiatric patients and is relatively safe and easy to administer. Nitrous oxide induces laughter which has been described as nature's epileptoid catharsis which one might surmise would be beneficial for depression. It also produces a central sympathetic stimulation similar to ECT and causes release of endogenous opioid peptides, which are potential candidates for the development of antidepressant drugs. Nitrous oxide is also associated with seizure like activity itself. Administration of nitrous oxide as a substitute for ECT is eminently feasible and could be given in a series of treatments similar to ECT therapy. PMID- 20006917 TI - Current treatment of cat bites to the hand and wrist. PMID- 20006918 TI - Predicting embolic potential during carotid angioplasty and stenting: analysis of captured particulate debris, ultrasound characteristics, and prior carotid endarterectomy. AB - INTRODUCTION: Extracranial carotid stenoses exhibit significant variance in embolic potential, with restenotic lesions having a particularly low propensity for embolization. This study sought to identify characteristics associated with increased generation of embolic debris during carotid angioplasty and stenting (CAS). METHODS: Captured particulate was available for analysis in 56 consecutive patients. Demographics were mean age, 74 years (range, 60-94 years); mean stenosis, 88% (range, 70%-99%); symptomatic, 27%; prior carotid endarterectomy (CEA), 27%; prior radiotherapy, 7%. Plaque echogenicity, heterogenicity, ulceration, and irregularity were assessed with B-mode duplex ultrasound analysis. Gray scale median (GSM) was calculated from normalized B-mode VHS video recordings. Calcification and degree of stenosis were determined angiographically. Captured particulate debris was evaluated for total number; number >200 microm, >500 microm, >1000 microm; mean and median size. Hematoxylin and eosin, trichrome, and von Kossa stains were used for histologic analysis of captured material. RESULTS: Restenotic carotid stenoses after prior CEA generated minimal embolic debris compared with primary stenoses. Four of 15 patients (27%) with restenotic lesions demonstrated embolic particles; all debris was <500 microm. All 41 patients with primary stenoses had some embolic debris; particulate size was >200 microm in 91%, >500 microm in 72%, and >1000 microm in 43%. In primary lesions, the number and size of captured particulate correlated with GSM and with the combined ultrasound findings of echogenicity, heterogenicity, and luminal irregularity/ulceration (P < .02, 95% confidence interval, 4.5-27.6). None of these ultrasound factors correlated independently with embolic particulate (P = NS). Patients aged >70 years exhibited more total particles (8.1 vs 2.3, P = .008) and increased mean particle size (370 vs 157 mum, P = .02). No significant correlation was observed between the number and size of captured embolic particulate and any other variable (stenosis percentage, prior radiotherapy, preprocedural symptoms, periprocedural symptoms, and calcification). Histologically, the embolic debris consisted of extensive amorphous, acellular proteinaceous material. Calcium debris in the embolic particulate was associated with heavily and moderately calcified lesions. CONCLUSIONS: Considerable variation exists in the number and size of embolic particles generated during CAS. Embolic potential is positively correlated with lesion GSM and the combination of lesion echogenicity, heterogenicity, and irregularity. Restenosis after prior CEA is associated with minimal embolic particulate generation, suggesting that embolic protection may not be necessary for CAS of restenotic lesions. PMID- 20006920 TI - Unexpected major role for venous stenting in deep reflux disease. AB - BACKGROUND: Treatment of chronic venous insufficiency (CVI) has largely focused on reflux. Minimally-invasive techniques to address superficial and perforator reflux have evolved, but correction of deep reflux continues to be challenging. The advent of intravascular ultrasound (IVUS) scan and minimally invasive venous stent technology have renewed interest in the obstructive component in CVI pathophysiology. The aim of this study is to assess stent-related and clinical outcomes following treatment by iliac venous stenting alone in limbs with a combination of iliac vein obstruction and deep venous reflux. METHODS: A total of 528 limbs in 504 patients, ranging in age from 15 to 87, underwent IVUS-guided iliac vein stent placement to correct obstruction over an 11-year period. The etiology of obstruction was nonthrombotic in 196 (37%), post-thrombotic in 285 (54%) limbs, and combined in 47 (9%). Clinical severity class of CEAP was C3 in 44%, C(4,5) in 27%, and C6 in 25% of stented limbs. Deep venous reflux was present in all limbs, associated with superficial and/or perforator reflux in 69%. Reflux was severe in 309/528 (59%) limbs (reflux multisegment score > or = 3) and 224/528 (42%) limbs had axial reflux. Venography and other functional tests had poor diagnostic sensitivity to detect obstruction, which was ultimately diagnosed by IVUS. The IVUS-guided iliac vein stenting was the only procedure performed and the associated reflux was left uncorrected. RESULTS: There was no mortality; morbidity was minor. Cumulative secondary stent patency was 88% at 5 years; no stent occlusions occurred in nonthrombotic limbs. Cumulative rates of limbs with healed active ulcers, freedom of ulcer recurrence in legs with healed ulcers (C5), and freedom from leg dermatitis at 5 years were 54%, 88%, and 81%, respectively. Cumulative rate of substantial improvement of pain and swelling at 5 years was 78% and 55%, respectively. Quality of life improved significantly. Reflux parameters did not deteriorate after stenting. CONCLUSION: Iliac venous stenting alone is sufficient to control symptoms in the majority of patients with combined outflow obstruction and deep reflux. Partial correction of the pathophysiology in limbs with multisystem or multilevel disease can provide substantial symptom relief. Percutaneous stent technology in concert with other minimally-invasive techniques to address superficial and/or perforator reflux offers such partial correction in limbs with advanced CVI and complex venous pathology. Open correction of obstruction or reflux is now required only infrequently as a "last resort". PMID- 20006921 TI - A phase Ib/IIa trial to evaluate the CCK2 receptor antagonist Z-360 in combination with gemcitabine in patients with advanced pancreatic cancer. AB - AIM: To evaluate the combination of the gastrin antagonist Z-360 and gemcitabine for advanced pancreatic cancer. METHODS: Previously untreated patients with PC were randomly allocated to Z-360 120 mg, 240 mg or placebo. Z-360/placebo was given on day -3 and gemcitabine 1000 mg/m(2) commenced on day 1 followed by Z-360 on day 2. Thereafter Z-360/placebo was given twice daily concurrently with standard dose of gemcitabine. Pharmacokinetics for both drugs was measured alone and in combination. Toxicity, response and quality of life were also recorded. RESULTS: Thirty-three patients with a median age of 62 years were randomised of which six had locally advanced disease and 26 had metastatic disease. Analysis of the area under the plasma concentration versus time curve (AUC), the maximum observed concentration (Cmax(obs)) and the time of the maximum observed concentration (Tmax(obs)) for Z-360, gemcitabine and 2,2-difluorodeoxyuridine (dFdU), could not exclude an effect on the systemic exposure to Z-360, gemcitabine and dFdU when co-administration of Z-360 and gemcitabine was compared with single agent administration. The most commonly reported adverse events were nausea, abdominal pain, vomiting and fatigue. At the end of the study, 62.5%, 25% and 60% had stable disease in the 120 mg, 240 mg and placebo group, respectively. A higher proportion of patients in Z-360 groups reported improvement in pain. CONCLUSIONS: Z-360 is safe and well tolerated when combined with gemcitabine. A Phase III trial is needed to determine whether the combination of Z-360 and gemcitabine is superior to gemcitabine alone in advanced PC. PMID- 20006922 TI - VEGF signalling inhibition-induced proteinuria: Mechanisms, significance and management. AB - Proteinuria is a dose-related side-effect occurring after inhibition of vascular endothelial growth factor (VEGF) signalling and may reflect severe glomerular damage. The inhibition of the VEGF signalling axis induces downexpression or suppression of nephrin, an important protein for the maintenance of the glomerular slit diaphragm, sometimes leading to nephritic syndrome and/or glomerular thrombotic microangiopathy, the main-associated kidney disease. A MEDLINE search was carried out using the following criteria: (1) all MEDLINE listings as of 01-01-2000 with abstracts; (2) English language; and (3) Humans. The following phrases were used to query the database: (proteinuria) AND (anti VEGF OR VEGF inhibition OR bevacizumab OR sunitinib OR sorafenib OR VEGF Trap OR axitinib OR pazopanib OR AZ 2171). The references of each article identified were carefully reviewed for additional reference. The incidence of mild and asymptomatic proteinuria ranges from 21% up to 63%, but heavy proteinuria has been reported in up to 6.5% of renal cell carcinoma patients. Although discontinuation of anti-VEGF agent induced significant reduction, persistence of proteinuria is common. Although angiotensinconverting-enzyme inhibitors and/or angiotensin receptor blockers seem to be preferred, no specific recommendation for an antiproteinuric agent can be made in this context because there are no controlled studies addressing the subject. Periodic monitoring of urinary protein should be carried out in anti-VEGF-treated patients and patients showing proteinuria need special referral to nephrologists. PMID- 20006923 TI - Adsorption behavior of heavy metals onto chemically modified sugarcane bagasse. AB - A new process for the xanthation of sugarcane (Saccharum officinarum) bagasse was investigated for the separation of cadmium, lead, nickel, zinc and copper from their aqueous solutions. Adsorption capacity of the charred xanthated sugarcane bagasse (CXSB) was found to be significantly more than the several biosorbents reported in the literatures. The modified material was characterized by FTIR and elemental analysis. The kinetics of sorption of the tested metals was fast, reaching equilibrium within 20-40 min. The maximum adsorption capacities evaluated in terms of mol/kg dry gel were 1.95 for Cd(II), 1.58 for Pb(II), 2.52 for Ni(II), 2.40 for Zn(II) and 2.91 for Cu(II), respectively. The high adsorption capacity and the kinetics results indicated that CXSB can be used as the selective adsorbent for the removal of these respective metal ions from wastewater. PMID- 20006924 TI - Pretreatment of poultry litter improves Bacillus thuringiensis-based biopesticides production. AB - Pretreated poultry litter was used in batch cultures for the production of Bacillus thuringiensis (Bt)-based biopesticide of lepidoptera- and diptera specific Cry1 and Cry2, diptera-specific Cry4Ba and Cry11Aa and coleoptera specific Cry3Aa toxins by Bt subsp. kurstaki 81, subsp. israelensis HD500 and subsp. tenebrionis 3203, respectively. Bt kurstaki 81 showed improved growth and produced more toxin in this medium as compared to other subspecies. Base and acid hydrolysis were tested as the methods of substrate pretreatment. The use of poultry litter pretreated with 2N HCl yielded 94% more bioinsecticidal protein than 2N NaOH-pretreated poultry litter when Bt kurstaki 81 was cultured. With appropriate pretreatment, poultry litter demonstrated potential as a valuable raw material for a low-cost complex medium to produce Bt-based biopesticides. PMID- 20006925 TI - Use of biogas fluid-soaked water hyacinth for cultivating Pleurotus geesteranus. AB - Experiments were carried out to test the viability of growing Pleurotus geesteranus on media containing varying amounts of crushed water hyacinth slices, which were soaked in pig farm biogas fluid and dried. The water hyacinth material was utilized to substitute sawdust in the media for mushroom cultivation. Mushroom fruiting bodies harvested were evaluated for yield, amino acid and heavy metal contents. Among the eight treatment groups, the greatest yield and highest amino acid content in the mushrooms were obtained when the proportions of water hyacinth and sawdust in the medium were equal. The concentrations of heavy metals, Hg, Pb and Cd, in most of the present mushroom samples did not exceed the maximum allowed levels according to the limits set forth by the food hygienic and safety regulations for edible mushrooms in China. The proposed waste utilization of water hyacinth could conceivably benefit the environment in various aspects including conservation of forest by reducing the demand on natural woods for mushroom production. PMID- 20006926 TI - Review: Continuous hydrolysis and fermentation for cellulosic ethanol production. AB - Ethanol made biologically from a variety of cellulosic biomass sources such as agricultural and forestry residues, grasses, and fast growing wood is widely recognized as a unique sustainable liquid transportation fuel with powerful economic, environmental, and strategic attributes, but production costs must be competitive for these benefits to be realized. Continuous hydrolysis and fermentation processes offer important potential advantages in reducing costs, but little has been done on continuous processing of cellulosic biomass to ethanol. As shown in this review, some continuous fermentations are now employed for commercial ethanol production from cane sugar and corn to take advantage of higher volumetric productivity, reduced labor costs, and reduced vessel down time for cleaning and filling. On the other hand, these systems are more susceptible to microbial contamination and require more sophisticated operations. Despite the latter challenges, continuous processes could be even more important to reducing the costs of overcoming the recalcitrance of cellulosic biomass, the primary obstacle to low cost fuels, through improving the effectiveness of utilizing expensive enzymes. In addition, continuous processing could be very beneficial in adapting fermentative organisms to the wide range of inhibitors generated during biomass pretreatment or its acid catalyzed hydrolysis. If sugar generation rates can be increased, the high cell densities in a continuous system could enable higher productivities and yields than in batch fermentations. PMID- 20006927 TI - A comparison of co-combustion characteristics of coal with wood and hydrothermally treated municipal solid waste. AB - In this work, thermogravimetric analysis was used to investigate the co combustion characteristics of wood and municipal solid waste (MSW) with Indian coal. Combustion characteristics like volatile release, ignition were studied. Wood presented an enhanced reaction rate reflecting its high volatile and low ash contents, while MSW enhanced ignition behavior of Indian coal. The results indicate that blending of both, wood and MSW improves devolatization properties of coal. Significant interaction was detected between wood and Indian coal, and reactivity of coal has improved upon blending with wood. On the other hand, MSW shows a good interaction with Indian coal leading to significant reduction in ignition temperature of coal and this effect was more pronounced with higher blending ratio of MSW. Hence MSW blending could more positively support the combustion of low quality Indian coal as compared to wood, due to its property of enhancement of ignition characteristics. PMID- 20006928 TI - Substrate specificity of family 5, 6, 7, 9, 12, and 45 endoglucanases. AB - Endoglucanases are important enzymes for biomass conversion and other industrial processes. Determining the specificity of endoglucanases from various glycoside hydrolase families is of interest for bioinformatic functional prediction and substrate-tailored enzyme development. To do so, we characterized approximately 30 endoglucanases from six glycoside hydrolase families. For p-nitrophenyl cellobioside and lactoside, only family 7 enzymes showed significant activity. For xyloglucan, both family 7 and 12 enzymes showed significant activity. For xylan and arabinoxylan, only family 7 enzymes showed significant activity. For mannan and galactomannan, both family 5 and 9 enzymes showed significant activity. The difference in specificity was preliminarily attributed mainly to the structural difference of the enzymes' active sites. For family 7 endoglucanases, difference in thermal stability might affect their performance in hydrolyzing various (hemi)cellulose substrates. Phylogenetic analysis on the subfamily distribution of family 5 endoglucanases (in relation with mannanases) suggested that their mannanase side-activity might be the remnant of an ancestral multi-function enzyme. Similar analysis was also made with the xyloglucanase or arabionxylans side-activity of family 12 and 7 endoglucanases. The apparent dependence of the specificity on family (primary/tertiary structure) might assist us in better understanding the structure-function relationship of the enzymes, and developing more versatile biocatalysts for the utilization of biomass. PMID- 20006929 TI - Discovery of novel taspine derivatives as antiangiogenic agents. AB - VEGFR-2 plays a critical role in vasculogenesis and inhibitors of VEGFR-2 could be used in the treatment of cancer. Taspine was one of the active ingredients screened by using an endothelial cell membrane chromatography and showed inhibition against VEGFR-2. In our research, we explored how the lactone ring and biphenyl scaffold in taspine influence its potent in vitro anticancer and antiangiogenesis activities. Accordingly, we report the design, synthesis, and preliminary evaluation of four novel taspine derivatives as VEGFR-2 inhibitors. The preliminary biological test showed that one of the compounds showed much better inhibitory activities against CACO-2 (IC(50)=52.5nM) and ECV304 (IC(50)=2.67nM) than taspine. This result enlarges the interest in ring-opened taspine derivative skeleton in the search of new antiangiogenesis agents. PMID- 20006930 TI - Femoral fixation pattern in cemented Oxford unicompartmental knee arthroplasty- an experimental cadaver study. AB - Femoral component loosening is a rare but serious complication in cemented Oxford unicompartmental knee arthroplasty (OUKA). In a cadaver study, OUKA was performed in 24 knees to evaluate the femoral fixation pattern. Due to the geometry of bone and implant, three different zones were identified. Complete cement mantles and good interdigitation were found in the spherical part of the implant (zone 1) and around the peg (zone 3), which implies that these are most important for implant fixation. The posterior plane facet (zone 2) is the weak point of the interface due to incompleteness of the cement mantle and a lack of interdigitation. This study suggests that the cancellous bone in zone 2 and the drill holes in sclerotic bone areas should be filled with cement and pressurised prior to component seating. PMID- 20006931 TI - Synthesis, characterization and antiglaucoma activity of a novel proton transfer compound and a mixed-ligand Zn(II) complex. AB - A novel proton transfer compound, pyridin-2-ylmethanaminium 2,4-dichloro-5 sulfamoylbenzoate (1), and a mixed-ligand Zn(II) complex, bis(2,4-dichloro-5 sulfamoylbenzoate)(2-aminomethylpyridine)aquazinc(II) monohydrate (2), have been synthesized from the same free ligands, which are 2,4-dichloro-5-sulfamoylbenzoic acid (Hsba) and 2-aminomethylpyridine (amp). They have been characterized by elemental, spectral ((1)H NMR, IR and UV-vis.) and thermal analyses. Additionally, magnetic measurement and single crystal X-ray diffraction technique were applied to compound 2. In the complex, Zn(II) ion exhibits a distorted octahedral configuration coordinated by O1 and O1(i) atoms of two mono dentante sba anions and N1, N2, N2(i) atoms of bidentante amp anion and a water molecule (O1w). The free ligands Hsba and amp, and the products 1 and 2, and acetazolamide (AAZ) as the control compound, were also evaluated for their in vitro inhibitor effects on human Carbonic Anhydrase isoenzymes (hCA I and hCA II) purified from erythrocyte cell by affinity chromatography for their hydratase and esterase activities. The IC(50) values of products 1 and 2 for hydratase activity are 0.26 and 0.13microM for hCA I and 0.30 and 0.15microM for hCA II, respectively. The IC(50) values of the same inhibitors for esterase activity are 0.32 and 0.045microM for hCA I and 0.29 and 0.23microM for hCA II, respectively. In relation to esterase activities, the inhibition equilibrium constants (K(i)) were also determined and found 0.25 and 0.058microM on hCA I and 0.22 and 0.24microM on hCA II for 1 and 2, respectively. The comparison of the inhibition studies of newly synthesized compounds 1 and 2 to parent compounds Hsba and amp and to AAZ indicated that 1 and 2 have effective inhibitory activity on hCA I and II, and might be used potential inhibitors. PMID- 20006932 TI - Assisted sonodynamic damage of bovine serum albumin by metronidazole under ultrasonic irradiation combined with photosensitive antitumor drug-Amsacrine. AB - By research, it was found that the Amsacrine (AMSA) can interact with bovine serum albumin (BSA). In this work, the AMSA was adopted as a sonosensitizer and the Metronidazole (MET) was used as a sensitizer to further damage BSA molecules under ultrasonic irradiation. It could be concluded that the damage degree of BSA molecules in the presence of AMSA and MET was more serious than in the presence of pure AMSA. That is, MET could aggravate the damage to BSA molecules under ultrasonic irradiation combined with AMSA. Meanwhile, the damage degree of BSA molecules was also influenced by some factors, such as ultrasonic irradiation time, MET concentration and solution acidity. In addition, the damage site of BSA molecules was estimated by synchronous fluorescence spectra. It was found that the tyrosine (Tyr) and tryptophan (Typ) residues were damaged almost averagely. Perhaps, these research results are of great significance for driving sonodynamic method to treat tumor in clinic application. PMID- 20006933 TI - Electronic storage of echocardiographic images: from clips to bits. PMID- 20006934 TI - Innovations in aortic disease: the ascending aorta and aortic arch. AB - Significant innovations have defined the approach to the proximal thoracic aorta. Aortic proteolysis predisposes to dissection and aneurysm. Losartan may prevent aortic root dilation in Marfan syndrome. The Loeys-Dietz syndrome mandates early aortic intervention. Because genetic aortopathies have a multicenter registry, further aortic molecular advances are likely. Acute intramural hematoma (IMH) may be due to aortic dissection with unrecognized microintimal tears. Type-A IMH is often a surgical emergency, whereas type-B IMH often requires medical management. Because preoperative ischemia predicts mortality in type-A dissection, it is logical to classify this disease by ischemic presentation. Because advanced age worsens the outcome in type-A dissection, transcatheter interventions should be urgently developed for this high-risk subgroup. Aortic arch repairs shorter than 45 minutes in duration are safely performed under deep hypothermic circulatory arrest with/without perfusion adjuncts. Bilateral antegrade cerebral perfusion (ACP) offers the best neuroprotection for complex repairs longer than 45 minutes. Axillary artery cannulation improves outcomes in proximal thoracic aortic procedures. Contralateral hemispheric ischemia is possible with unilateral ACP because cross-cerebral perfusion may be inadequate. Arch repair with ACP and moderate HCA is safe and effective and represents a research opportunity for pharmacologic ischemic preconditioning. Antegrade thoracic aortic stenting for DeBakey 1 dissection thromboses the distal false lumen to improve long-term aortic outcomes. Endovascular arch repair is feasible and may soon be done off pump. These described innovations have collectively ushered in a paradigm shift in diseases affecting the ascending aorta and aortic arch. PMID- 20006935 TI - Prognostic impact of Chagas' disease in patients awaiting heart transplantation. AB - BACKGROUND: The role of Chagas' etiology of chronic heart failure in predicting patient outcomes while awaiting heart transplantation is unknown. Accordingly, in this study we compare outcomes in Chagas' disease with non-Chagas'-disease related advanced heart failure among patients on the waiting list for heart transplantation. METHODS: We reviewed the clinical outcomes of 103 consecutive patients with chronic heart failure listed for heart transplantation from August 2000 to January 2008 at a single institution. Forty-six (44%) patients were diagnosed with Chagas' disease on the basis of positive serology. A Cox proportional hazards model was used to establish independent predictors of mortality, whereas competing risk analysis was used to estimate time-related prevalence of death and heart transplantation in Chagas' disease and non-Chagas' disease patients. RESULTS: In the multivariate model, inotropic support (p < 0.0005; hazard ratio = 5.96; 95% confidence interval [CI] 2.41 to 14.71) and Chagas' disease etiology of heart failure (p = 0.02; hazard ratio = 2.27; 95% CI 1.14 to 4.52) were retained as independent predictors of mortality. Prevalence of death at 100 days after listing was 30% in Chagas' disease and 16% in non-Chagas' disease patients (p = 0.02), despite no difference in the competing rates of transplantation (30% in Chagas' and 37% in non-Chagas' patients, p = 0.5). CONCLUSIONS: Chagas' disease etiology serves as an independent predictor of mortality in patients listed for heart transplantation, with a worse outcome when compared with non-Chagas' disease patients. PMID- 20006936 TI - Role of psychological factors in the clinical course of heart transplant patients. PMID- 20006937 TI - Salmon poisoning disease in dogs: a satisfying diagnosis. PMID- 20006938 TI - Ultrasound-assisted microwave preparation of Ag-doped CdS nanoparticles. AB - Ag-doped CdS nanoparticles were synthesized by an ultrasound-assisted microwave synthesis method. The X-ray diffraction patterns reveal a structural evolution from cubic to hexagonal with increasing molar ratios of Ag(+)/Cd(2+) from 0% to 5%. It shows that the Ag-doped hexagonal CdS nanoparticles are polycrystal. The X ray photoelectron spectroscopy of the CdS nanoparticles doping with 5% Ag(+) shows that the doped Ag in CdS is metallic. Simultaneously, the characteristic Raman peaks of the CdS nanoparticles enhance with increasing Ag(+) concentrations. The photocatalytic activity of different Ag-doped samples show a reasonable change due to different ratios of Ag which doped into CdS. PMID- 20006939 TI - Kinetic study on the effect of ultrasound on lipase-catalyzed hydrolysis of soy oil: Study of the interfacial area and the initial rates. AB - Comparative studies of lipase-catalyzed hydrolysis of soy oil takes place at the interface between the oil and the aqueous solution in solvent-free system were carried out in shaking bath and in ultrasonic bath. The interfacial area between the oil phase and the aqueous phase influences the rate of hydrolysis. Compared to shaking bath, ultrasonic shaking was found to be a more effective procedure to disperse the oil in water. Larger interfacial area and smaller drop size could be obtained in ultrasonic bath. The initial rate of hydrolysis was increased with the increasing of interfacial area. In ultrasonic bath, the highest initial rate of reaction was obtained with the oil volume fraction of 0.7, which was in accord to the highest interfacial area obtained with the oil volume fraction of 0.7, when the ultrasonic power was 1.64W/cm(2). The higher initial rate of hydrolysis was 161mol/m(3)min. The ultrasonic bath was a useful way to disperse soy oil in water to obtain a larger interfacial area, which caused the higher initial rate of soy oil hydrolysis in the solvent-free system. PMID- 20006940 TI - Effect of in vitroexpanded CD4(+)CD25(+)Foxp3(+) regulatory T cell therapy combined with lymphodepletion in murine skin allotransplantation. AB - A promising approach for preventing allograft rejection involves shifting the balance between cytopathic and regulatory T cells to dominance of the latter cell type. Nonspecific lymphodepletion was conducted by administration of depleting anti-CD4 and anti-CD8 antibodies to reduce effector T cells and adoptive transfer of ex vivo-expanded host Treg cells by stimulation with donor dendritic cells to augment the Treg cell compartment. Evaluation of an MHC-mismatched skin allograft model revealed that combined therapy with these two protocols consistently induced modest prolongation of allograft survival, although all skin grafts were eventually rejected. The administration of IL-2/anti-IL-2 complexes significantly improved the efficacy of combination therapy via promoting the expansion of adoptively transferred Treg cells as well as endogenous recipient Treg cells. We conclude that Treg cell therapy combined with lymphodepletion is of practical benefit for the control of allograft rejection, but requires supplementary measures to promote immune tolerance. PMID- 20006941 TI - The TMEM16 protein family: a new class of chloride channels? AB - Cl(-) channels play important roles in many physiological processes, including transepithelial ion absorption and secretion, smooth and skeletal muscle contraction, neuronal excitability, sensory perception, and cell volume regulation. The molecular identity of many types of Cl(-) channels is still unknown. Recently, three research groups have arrived independently at the identification of TMEM16A (also known as anoctamin-1) as a membrane protein strongly related to the activity of Ca(2+)-activated Cl(-) channels (CaCCs). Site specific mutagenesis of TMEM16A alters the properties of the channels, thus suggesting that TMEM16A forms, at least in part, the CaCC. TMEM16A is a member of a family that includes nine other membrane proteins. All TMEM16 proteins have a similar structure, with eight putative transmembrane domains and cytosolic amino- and carboxy-termini. TMEM16B expression also evokes the appearance of CaCCs, but with biophysical characteristics (voltage dependence, unitary conductance) different from those associated to TMEM16A. The roles of the other TMEM16 proteins are still unknown. The study of TMEM16 proteins may lead to identification of novel molecular mechanisms underlying ion transport and channel gating by voltage and Ca(2+). PMID- 20006942 TI - A biomechanical triphasic approach to the transport of nondilute solutions in articular cartilage. AB - Biomechanical models for biological tissues such as articular cartilage generally contain an ideal, dilute solution assumption. In this article, a biomechanical triphasic model of cartilage is described that includes nondilute treatment of concentrated solutions such as those applied in vitrification of biological tissues. The chemical potential equations of the triphasic model are modified and the transport equations are adjusted for the volume fraction and frictional coefficients of the solutes that are not negligible in such solutions. Four transport parameters, i.e., water permeability, solute permeability, diffusion coefficient of solute in solvent within the cartilage, and the cartilage stiffness modulus, are defined as four degrees of freedom for the model. Water and solute transport in cartilage were simulated using the model and predictions of average concentration increase and cartilage weight were fit to experimental data to obtain the values of the four transport parameters. As far as we know, this is the first study to formulate the solvent and solute transport equations of nondilute solutions in the cartilage matrix. It is shown that the values obtained for the transport parameters are within the ranges reported in the available literature, which confirms the proposed model approach. PMID- 20006943 TI - Calcium involvement in regulation of neuronal bursting in disinhibited neuronal networks: insights from calcium studies in a spherical cell model. AB - Cytosolic calcium is involved in the regulation of many intracellular processes. Intracellular calcium may therefore potentially affect the behavior of both single neurons and synaptically connected neuronal assemblies. In computer model studies, we investigated calcium dynamics in spherical neurons during periods of recurrent neuronal bursting that were simulated in a disinhibited neuronal network. The model takes into account calcium influx via voltage-gated calcium channels, extrusion through the cell membrane, and binding to two different buffers representing fixed and mobile endogenous calcium buffers. Throughout the duration of the simulated recurrent neuronal bursting, the concentration of free fixed buffers shows a hyperbolic decrease in time at a rate that is not uniform inside a neuron. Recurrent calcium influxes associated with bursting lead to the formation of gradients in the concentration of the fixed buffer in the radial direction, and are accompanied by the redistribution of mobile buffers acting to compensate for these gradients. Simulated intracellular calcium transients have a slow component characterized by a gradual increase in the calcium baseline level that reaches a plateau 120-200 s after the onset of recurrent bursting. Using this model, we demonstrate what we believe is a novel mechanism of regulation of network excitability that occurs in conditions of prolonged and recurrent neuronal bursting in disinhibited networks. This mechanism is expressed via interaction of calcium clearance systems inside neurons with calcium-dependent potassium regulation of neuronal excitability in membranes. This is a network phenomenon because it arises largely by synaptic interactions. Therefore, it can serve as a network safety mechanism to prevent excessive and uncontrolled neuronal firing resulting from the lack of inhibition or after acute suppression of the inhibitory drive. PMID- 20006944 TI - Combining laser microsurgery and finite element modeling to assess cell-level epithelial mechanics. AB - Laser microsurgery and finite element modeling are used to determine the cell level mechanics of the amnioserosa-a morphogenetically crucial epithelium on the dorsal surface of fruit fly embryos (Drosophila melanogaster). In the experiments, a tightly focused laser ablates a subcellular hole (1 microm in diameter) that passes clean through the epithelium. The surrounding cells recoil from the wound site with a large range of initial recoil velocities. These depend on the embryo's developmental stage and the subcellular wound site. The initial recoil (up to 0.1 s) is well reproduced by a base finite element model, which assumes a uniform effective viscosity inside the cells, a constant tension along each cell-cell boundary, and a large, potentially anisotropic, far-field stress- one that far exceeds the stress equivalent of the cell-edge tensions. After 0.1 s, the experimental recoils slow dramatically. This observation can be reproduced by adding viscoelastic rods along cell edges or as a fine prestressed mesh parallel to the apical and basal membranes of the cell. The mesh also reproduces a number of double-wounding experiments in which successive holes are drilled in a single cell. PMID- 20006945 TI - A novel method to quantify contribution of channels and transporters to membrane potential dynamics. AB - The action potential, once triggered in ventricular or atrial myocytes, automatically proceeds on its time course or is generated spontaneously in sinoatrial node pacemaker cells. It is induced by complex interactions among such cellular components as ion channels, transporters, intracellular ion concentrations, and signaling molecules. We have developed what is, to our knowledge, a new method using a mathematical model to quantify the contribution of each cellular component to the automatic time courses of the action potential. In this method, an equilibrium value, which the membrane potential is approaching at a given moment, is calculated along the time course of the membrane potential. The calculation itself is based on the time-varying conductance and the reversal potentials of individual ion channels and electrogenic ion transporters. Since the equilibrium potential moves in advance of the membrane potential change, we refer to it as the lead potential, V(L). The contribution of an individual current was successfully quantified by comparing dV(L)/dt before and after fixing the time-dependent change of a component of interest, such as the variations in the open probability of a channel or the turnover rate of an ion transporter. In addition to the action potential, the lead-potential analysis should also be applicable in all types of membrane excitation in many different kinds of cells. PMID- 20006946 TI - Phosphorylation facilitates the integrin binding of filamin under force. AB - Filamins are actin binding proteins that contribute to cytoskeletal integrity and biochemical scaffolds during mechanochemical signal transductions. Structurally, human filamins are dimers composed of an actin-binding domain with 24 immunoglobulin (Ig)-like repeats. In this study, we focus on the recently solved high-resolution crystal structure of Ig-like repeats 19-21 of filamin-A (IgFLNa R19-R21). IgFLNa-R19-21 is of marked importance because it contains the binding site for integrins and facilitates the dynamic ability of filamin-A to communicate with the extracellular environment. However, the structure of filamin A shows an interesting domain arrangement where the integrin binding site on IgFLNa-R21 is hindered sterically by IgFLNa-R20. Thus, a number of hypotheses on the regulation of filamin-A exist. Using molecular dynamics simulations we evaluated the effects of two primary regulators of filamin-A, force and phosphorylation. We find that a tensile force of 40 pN is sufficient to initiate the partial removal of the autoinhibition on the integrin binding site of IgFLNa R21. Force coupled to phosphorylation at Ser(2152), however, affords complete dissociation of autoinhibition with a decreased force requirement. Phosphorylation seems to decrease the threshold for removing the IgFLNa-R20 beta strand inhibitor within 300 ps with 40 pN tensile force. Furthermore, the molecular dynamic trajectories illustrate phosphorylation of Ser(2152) without force is insufficient to remove autoinhibition. We believe the results of this study implicate filamin-A as a tunable mechanosensor, where its sensitivity can be modulated by the degree of phosphorylation. PMID- 20006947 TI - Anchorage of vinculin to lipid membranes influences cell mechanical properties. AB - The focal adhesion protein vinculin (1066 residues) can be separated into a 95 kDa head and a 30-kDa tail domain. Vinculin's lipid binding sites localized on the tail, helix 3 (residues 944-978) and the unstructured C-terminal arm (residues 1052-1066, the so-called lipid anchor), influence focal adhesion turnover and are important for cell migration and adhesion. Using magnetic tweezers, we characterized the cell mechanical behavior in mouse embryonic fibroblast (MEF)-vin(-/-) cells transfected with EGFP-linked-vinculin deficient of the lipid anchor (vinDeltaC, residues 1-1051). MEF-vinDeltaC cells incubated with fibronectin-coated paramagnetic beads were less stiff, and more beads detached during these experiments compared to MEF-rescue cells. Cells expressing vinDeltaC formed fewer focal contacts as determined by confocal microscopy. Two dimensional traction measurements showed that MEF-vinDeltaC cells generate less force compared to rescue cells. Attenuated traction forces were also found in cells that expressed vinculin with point mutations (R1060 and K1061 to Q) of the lipid anchor that impaired lipid binding. However, traction generation was not diminished in cells that expressed vinculin with impaired lipid binding caused by point mutations on helix 3. Mutating the src-phosphorylation site (Y1065 to F) resulted in reduced traction generation. These observations show that both the lipid binding and the src-phosphorylation of vinculin's C-terminus are important for cell mechanical behavior. PMID- 20006948 TI - Flip-flop-induced relaxation of bending energy: implications for membrane remodeling. AB - Cellular and organellar membranes are dynamic materials that underlie many aspects of cell biology. Biological membranes have long been thought of as elastic materials with respect to bending deformations. A wealth of theory and experimentation on pure phospholipid membranes provides abundant support for this idea. However, biological membranes are not composed solely of phospholipids- they also incorporate a variety of amphiphilic molecules that undergo rapid transbilayer flip-flop. Here we describe several experimental systems that demonstrate deformation-induced molecular flip-flop. First we use a fluorescence assay to track osmotically controlled membrane deformation in single component fatty acid vesicles, and show that the relaxation of the induced bending stress is mediated by fatty acid flip-flop. We then look at two-component phospholipid/cholesterol composite vesicles. We use NMR to show that the steady state rate of interleaflet diffusion of cholesterol is fast relative to biological membrane remodeling. We then use a Forster resonance energy transfer assay to detect the transbilayer movement of cholesterol upon deformation. We suggest that our results can be interpreted by modifying the area difference elasticity model to account for the time-dependent relaxation of bending energy. Our findings suggest that rapid interleaflet diffusion of cholesterol may play a role in membrane remodeling in vivo. We suggest that the molecular characteristics of sterols make them evolutionarily preferred mediators of stress relaxation, and that the universal presence of sterols in the membranes of eukaryotes, even at low concentrations, reflects the importance of membrane remodeling in eukaryotic cells. PMID- 20006949 TI - Extensibility of the extended tail domain of processive and nonprocessive myosin V molecules. AB - Myosin V is a single-molecule motor that moves organelles along actin. When myosin V pulls loads inside the cell in a highly viscous environment, the force on the motor is unlikely to be constant. We propose that the tether between the single-molecule motor and the cargo (i.e., the extended tail domain of the molecule) must be able to absorb the sudden mechanical motions of the motor and allow smooth relaxation of the motion of the cargo to a new position. To test this hypothesis, we compared the elastic properties of the extended tail domains of processive (mouse myosin Va) and nonprocessive (Drosophila myosin V) molecular motors. The extended tail domain of these myosins consists of mechanically strong coiled-coil regions interspersed with flexible loops. In this work we explored the mechanical properties of coiled-coil regions using atomic force microscopy. We found that the processive and nonprocessive coiled-coil fragments display different unfolding patterns. The unfolding of coiled-coil structures occurs much later during the atomic force microscopy stretch cycle for processive myosin Va than for nonprocessive Drosophila myosin V, suggesting that this elastic tether between the cargo and motor may play an important role in sustaining the processive motions of this single-molecule motor. PMID- 20006950 TI - Quick shear-flow alignment of biological filaments for X-ray fiber diffraction facilitated by methylcellulose. AB - X-ray fiber diffraction is one of the most useful methods for examining the structural details of live biological filaments under physiological conditions. To investigate biologically active or labile materials, it is crucial to finish fiber alignment within seconds before diffraction analysis. However, the conventional methods, e.g., magnetic field alignment and low-speed centrifugations, are time-consuming and not very useful for such purposes. Here, we introduce a new alignment method using a rheometer with two parallel disks, which was applied to observe fiber diffractions of axonemes, tobacco mosaic tobamovirus, and microtubules. We found that fibers were aligned within 5 s by giving high shear flow (1000-5000 s(-1)) to the medium and that methylcellulose contained in the medium (approximately 1%) was essential to the accomplishment of uniform orientation with a small angular deviation (<5 degrees). The new alignment method enabled us to execute structure analyses of axonemes by small angle x-ray diffraction. Since this method was also useful for the quick alignment of purified microtubules, as well as tobacco mosaic tobamovirus, we expect that we can apply it to the structural analysis of many other biological filaments. PMID- 20006952 TI - Nucleosome assembly depends on the torsion in the DNA molecule: a magnetic tweezers study. AB - We have used magnetic tweezers to study nucleosome assembly on topologically constrained DNA molecules. Assembly was achieved using chicken erythrocyte core histones and histone chaperone protein Nap1 under constant low force. We have observed only partial assembly when the DNA was topologically constrained and much more complete assembly on unconstrained (nicked) DNA tethers. To verify our hypothesis that the lack of full nucleosome assembly on topologically constrained tethers was due to compensatory accumulation of positive supercoiling in the rest of the template, we carried out experiments in which we mechanically relieved the positive supercoiling by rotating the external magnetic field at certain time points of the assembly process. Indeed, such rotation did lead to the same nucleosome saturation level as in the case of nicked tethers. We conclude that levels of positive supercoiling in the range of 0.025-0.051 (most probably in the form of twist) stall the nucleosome assembly process. PMID- 20006951 TI - Binding of the bacteriophage P22 N-peptide to the boxB RNA motif studied by molecular dynamics simulations. AB - Protein-RNA interactions are important for many cellular processes. The Nut utilization site (N)-protein of bacteriophages contains an N-terminal arginine rich motif that undergoes a folding transition upon binding to the boxB RNA hairpin loop target structure. Molecular dynamics simulations were used to investigate the dynamics of the P22 N-peptide-boxB complex and to elucidate the energetic contributions to binding. In addition, the free-energy changes of RNA and peptide conformational adaptation to the bound forms, as well as the role of strongly bound water molecules at the peptide-RNA interface, were studied. The influence of peptide amino acid substitutions and the salt dependence of interaction were investigated and showed good agreement with available experimental results. Several tightly bound water molecules were found at the RNA binding interface in both the presence and absence of N-peptide. Explicit consideration of the waters resulted in shifts of calculated contributions during the energetic analysis, but overall similar binding energy contributions were found. Of interest, it was found that the electrostatic field of the RNA has a favorable influence on the coil-to-alpha-helix transition of the N-peptide already outside of the peptide-binding site. This result may have important implications for understanding peptide-RNA complex formation, which often involves coupled folding and association processes. It indicates that electrostatic interactions near RNA molecules can lead to a shift in the equilibrium toward the bound form of an interacting partner before it enters the binding pocket. PMID- 20006953 TI - Force-driven separation of short double-stranded DNA. AB - Short double-stranded DNA is used in a variety of nanotechnological applications, and for many of them, it is important to know for which forces and which force loading rates the DNA duplex remains stable. In this work, we develop a theoretical model that describes the force-dependent dissociation rate for DNA duplexes tens of basepairs long under tension along their axes ("shear geometry"). Explicitly, we set up a three-state equilibrium model and apply the canonical transition state theory to calculate the kinetic rates for strand unpairing and the rupture-force distribution as a function of the separation velocity of the end-to-end distance. Theory is in excellent agreement with actual single-molecule force spectroscopy results and even allows for the prediction of the rupture-force distribution for a given DNA duplex sequence and separation velocity. We further show that for describing double-stranded DNA separation kinetics, our model is a significant refinement of the conventionally used Bell Evans model. PMID- 20006954 TI - NMR and MD investigations of human galectin-1/oligosaccharide complexes. AB - The specific recognition of carbohydrates by lectins plays a major role in many cellular processes. Galectin-1 belongs to a family of 15 structurally related beta-galactoside binding proteins that are able to control a variety of cellular events, including cell cycle regulation, adhesion, proliferation, and apoptosis. The three-dimensional structure of galectin-1 has been solved by x-ray crystallography in the free form and in complex with various carbohydrate ligands. In this work, we used a combination of two-dimensional NMR titration experiments and molecular-dynamics simulations with explicit solvent to study the mode of interaction between human galectin-1 and five galactose-containing ligands. Isothermal titration calorimetry measurements were performed to determine their affinities for galectin-1. The contribution of the different hexopyranose units in the protein-carbohydrate interaction was given particular consideration. Although the galactose moiety of each oligosaccharide is necessary for binding, it is not sufficient by itself. The nature of both the reducing sugar in the disaccharide and the interglycosidic linkage play essential roles in the binding to human galectin-1. PMID- 20006955 TI - The structural basis for the prevention of nonsteroidal antiinflammatory drug induced gastrointestinal tract damage by the C-lobe of bovine colostrum lactoferrin. AB - Nonsteroidal antiinflammatory drugs (NSAIDs), due to their good efficacy in the treatment of pain, inflammation, and fever, are among the most prescribed class of medicines in the world. The main drawback of NSAIDs is that they induce gastric complications such as peptic ulceration and injury to the intestine. Four NSAIDs, indomethacin, diclofenac, aspirin, and ibuprofen were selected to induce gastropathy in mouse models. It was found that the addition of C-terminal half of bovine lactoferrin (C-lobe) reversed the NSAID-induced injuries to the extent of 47-70% whereas the coadministration of C-lobe prevented it significantly. The C lobe was prepared proteolytically using serine proteases. The binding studies of C-lobe with NSAIDs showed that these compounds bind to C-lobe with affinities ranging from 2.6 to 4.8 x 10(-4) M. The complexes of C-lobe were prepared with the above four NSAIDs. All four complexes were crystallized and their detailed three-dimensional structures were determined using x-ray crystallographic method. The structures showed that all the four NSAID molecules bound to C-lobe at the newly identified ligand binding site in C-lobe that is formed involving two alpha helices, alpha10 and alpha11. The ligand binding site is separated from the well known iron binding site by the longest and the most stable beta-strand, betaj, in the structure. Similar results were also obtained with the full length lactoferrin molecule. This novel, to our knowledge, binding site in C-lobe of lactoferrin shows a good complementarity for the acidic and lipophilic compounds such as NSAIDs. We believe this indicates that C-lobe of lactoferrin can be exploited for the prevention of NSAID-induced gastropathy. PMID- 20006956 TI - Molecular modeling of the misfolded insulin subunit and amyloid fibril. AB - Insulin, a small hormone protein comprising 51 residues in two disulfide-linked polypeptide chains, adopts a predominantly alpha-helical conformation in its native state. It readily undergoes protein misfolding and aggregates into amyloid fibrils under a variety of conditions. Insulin is a unique model system in which to study protein fibrillization, since its three disulfide bridges are retained in the fibrillar state and thus limit the conformational space available to the polypeptide chains during misfolding and fibrillization. Taking into account this unique conformational restriction, we modeled possible monomeric subunits of the insulin amyloid fibrils using beta-solenoid folds, namely, the beta-helix and beta-roll. Both models agreed with currently available biophysical data. We performed molecular dynamics simulations, which allowed some limited insights into the relative structural stability, suggesting that the beta-roll subunit model may be more stable than the beta-helix subunit model. We also constructed beta-solenoid-based insulin fibril models and conducted fiber diffraction simulation to identify plausible fibril architectures of insulin amyloid. A comparison of simulated fiber diffraction patterns of the fibril models to the experimental insulin x-ray fiber diffraction data suggests that the model fibers composed of six twisted beta-roll protofilaments provide the most reasonable fit to available experimental diffraction patterns and previous biophysical studies. PMID- 20006957 TI - Learning rates and states from biophysical time series: a Bayesian approach to model selection and single-molecule FRET data. AB - Time series data provided by single-molecule Forster resonance energy transfer (smFRET) experiments offer the opportunity to infer not only model parameters describing molecular complexes, e.g., rate constants, but also information about the model itself, e.g., the number of conformational states. Resolving whether such states exist or how many of them exist requires a careful approach to the problem of model selection, here meaning discrimination among models with differing numbers of states. The most straightforward approach to model selection generalizes the common idea of maximum likelihood--selecting the most likely parameter values--to maximum evidence: selecting the most likely model. In either case, such an inference presents a tremendous computational challenge, which we here address by exploiting an approximation technique termed variational Bayesian expectation maximization. We demonstrate how this technique can be applied to temporal data such as smFRET time series; show superior statistical consistency relative to the maximum likelihood approach; compare its performance on smFRET data generated from experiments on the ribosome; and illustrate how model selection in such probabilistic or generative modeling can facilitate analysis of closely related temporal data currently prevalent in biophysics. Source code used in this analysis, including a graphical user interface, is available open source via http://vbFRET.sourceforge.net. PMID- 20006958 TI - New insight into erythrocyte through in vivo surface-enhanced Raman spectroscopy. AB - The article presents a noninvasive approach to the study of erythrocyte properties by means of a comparative analysis of signals obtained by surface enhanced Raman spectroscopy (SERS) and resonance Raman spectroscopy (RS). We report step-by-step the procedure for preparing experimental samples containing erythrocytes in their normal physiological environment in a mixture of colloid solution with silver nanoparticles and the procedure for the optimization of SERS conditions to achieve high signal enhancement without affecting the properties of living erythrocytes. By means of three independent techniques, we demonstrate that under the proposed conditions a colloid solution of silver nanoparticles does not affect the properties of erythrocytes. For the first time to our knowledge, we describe how to use the SERS-RS approach to study two populations of hemoglobin molecules inside an intact living erythrocyte: submembrane and cytosolic hemoglobin (Hb(sm) and Hb(c)). We show that the conformation of Hb(sm) differs from the conformation of Hb(c). This finding has an important application, as the comparative study of Hb(sm) and Hb(c) could be successfully used in biomedical research and diagnostic tests. PMID- 20006959 TI - A femtosecond visible/visible and visible/mid-infrared transient absorption study of the light harvesting complex II. AB - Light harvesting complex II (LHCII) is the most abundant protein in the thylakoid membrane of higher plants and green algae. LHCII acts to collect solar radiation, transferring this energy mainly toward photosystem II, with a smaller amount going to photosystem I; it is then converted into a chemical, storable form. We performed time-resolved femtosecond visible pump/mid-infrared probe and visible pump/visible probe absorption difference spectroscopy on purified LHCII to gain insight into the energy transfer in this complex occurring in the femto picosecond time regime. We find that information derived from mid-infrared spectra, together with structural and modeling information, provides a unique visualization of the flow of energy via the bottleneck pigment chlorophyll a604. PMID- 20006960 TI - Subdiffraction-limit two-photon fluorescence microscopy for GFP-tagged cell imaging. AB - We report applications of two-photon excitation fluorescence (2PEF) microscopy with subdiffraction-limit resolution for green-fluorescent-protein-tagged cell imaging. The microscope integrates 2PEF microscopy and stimulated emission depletion microscopy in one microscope that has the benefits of both techniques: intrinsic three-dimensional resolution, confined photobleaching, and subdiffraction-limit resolution. The subdiffraction-limit resolution was demonstrated by resolving green-fluorescent-protein-tagged caveolar vesicles located within a distance shorter than the diffraction limit of a regular 2PEF microscope, which is approximately 250 nm even with the best optics. The full width at half-maximum of the effective point-spread function for the 2PEF microscope was estimated to be approximately 54 nm. PMID- 20006961 TI - Renoprotective properties of pirfenidone in subtotally nephrectomized rats. AB - Renal fibrosis is the final common pathway of chronic kidney disease, and its progression predicts the degree of renal dysfunction. We investigated the renoprotective properties of pirfenidone in a remnant kidney model of chronic renal failure to determine its pharmacological potency compared to enalapril. Five-sixths nephrectomized rats were fed diet containing pirfenidone (approximately 700mg/kg/day) for 8weeks. Pirfenidone steadily inhibited the progression of proteinuria, but not to a significant degree. Pirfenidone prevented the elevation of plasma creatinine and blood urea nitrogen. At the end of the experiment, pirfenidone had reduced systolic blood pressure by means of its renoprotective effect. In a histological study, pirfenidone improved interstitial fibrosis in the renal cortex. These effects were supported by the suppression of the expression of TGF-beta and fibronectin in the mRNA of the kidney. In contrast, pirfenidone had little effect on the expression of alpha smooth muscle actin, which is one of the proteins responsible for epithelial mesenchymal transition. This property was confirmed by the TGF-beta-induced transdifferentiation observed in cultured normal rat kidney tubular epithelial NRK52E cells. These results suggest that pirfenidone improves the progression of chronic renal failure via its antifibrotic action, although pirfenidone has less effective TGF-beta-induced epithelial to mesenchymal transdifferentiation. PMID- 20006962 TI - Selective MyD88-dependent pathway inhibition by the cyanobacterial natural product malyngamide F acetate. AB - In response to evolutionary selective pressure, prokaryotes have developed a rich array of secondary metabolites, some of which may be inhibitory to the innate immune system and the inflammatory response in vertebrates. We utilized the RAW264.7 macrophage cell line stimulated with LPS in a nitric oxide (NO) assay to screen for compounds with immunomodulatory activities from a library of marine natural products, and found that the malyngamide structure class, found commonly in the marine cyanobacterium Lyngbya majuscula, has potent activity. Several of the malyngamides were found to possess IC50 values of 5.4-18microM. Malyngamide F acetate exhibited strong concentration-dependent anti-inflammatory activity in the NO assay with an IC50 of 7.1microM and with no cytotoxicity at the concentrations tested. Subsequent real-time PCR of selected genes revealed a unique cytokine profile after LPS stimulation (TLR4) with decreased expression of iNOS, IL-1beta, IL-6, and IL-10, but increased TNF-alpha expression. Additional experiments utilizing CpG and Poly I:C stimulation to selectively activate the MyD88-dependent and -independent pathways via TLR9 and TLR3 substantiated the finding that malyngamide F acetate selectively inhibits the MyD88-dependent pathway. To our knowledge, this is the first report of a natural product inhibiting the MyD88-dependent pathway. PMID- 20006963 TI - Effects and mechanisms of aloperine on 2, 4-dinitrofluorobenzene-induced allergic contact dermatitis in BALB/c mice. AB - Allergic contact dermatitis is a prototypic T-cell-mediated cutaneous inflammatory response. Multiple cell types, inflammatory mediators and cytokines are involved in the regulation of immunologic and inflammatory processes in allergic contact dermatitis. Aloperine is an isolated alkaloid found in the plant of Sophora alopecuroides L. It has been clinically proved effective in China for a long time for skin inflammatory diseases such as allergic contact dermatitis. However, the mechanism of aloperine on allergic contact dermatitis is largely unknown. Therefore, the aim of this study was to investigate the effect of aloperine on 2, 4-dinitrofluorobenzene (DNFB)-induced allergic contact dermatitis in BALB/c mice and the possible underlying mechanisms. The results showed that topical application of DNFB on the ear provoked typical allergic contact dermatitis with ear swelling and ear erythema in BALB/c mice. Treatments with 1% aloperine suppressed DNFB-induced increase in ear thickness and ear erythema. Moreover, 1% aloperine treatment significantly decreased the up-regulated mRNA and protein levels of tumor necrosis factor-alpha (TNF-alpha), interleukin-1beta (IL-1beta) and interleukin-6 (IL-6) induced by DNFB in ear biopsy homogenates. Our findings suggest that aloperine greatly improves the DNFB-induced allergic contact dermatitis in mice. The therapeutic mechanism might be related to the reduction of TNF-alpha, IL-1beta and IL-6 production induced by DNFB. PMID- 20006964 TI - Do odors from different cats induce equivalent unconditioned and conditioned responses in rats? AB - Since cats are predators of rats and mice, interest in the consequences of exposure to cat-associated odors has increased in the last decade, particularly regarding the development of putative animal models of post-traumatic stress disorder (PTSD). Although in the literature there are some comments on the variability of the effects depending on the individual cat used, there are no reports on this subject. In the present study, we demonstrated, using male Sprague-Dawley rats and cloths impregnated with fur/skin odors from three different cats (one ovariectomized female and two intact males), that the unconditioned endocrine (release of corticosterone) and behavioral (inhibition of activity and avoidance of the cloth area) responses to the presence of the cat odors were statistically significant and similar among the three cats. The conditioned behavioral response to a clean cloth, studied 7 days after the initial exposure to odors, was also evident with the three cats, with minor differences among them. In contrast, only the fur odor from one of the male cats clearly increased anxiety-like behavior in the elevated plus-maze 6 days later. These data indicate that: (i) evaluation of unconditioned and conditioned responses to cat odors does not appear to strongly predict long-lasting increases in anxiety-like behavior; and, therefore, both types of responses are partially dissociated; and (ii) differences among cats mainly affected the induction of long-lasting changes in anxiety-like behavior. The ultimate reasons for these differences are not known, but their characterization is critical for a proper understanding of putative PTSD models. PMID- 20006965 TI - Medial amygdala lesions differentially influence stress responsivity and sensorimotor gating in rats. AB - BACKGROUND: The amygdala is involved in the coordination of stress but is also an important gatekeeper involved in the regulation of vigilance. The amygdala is structurally complex, consisting of several nuclei with specific functions in the affective response to environmental stimuli. There are indications that the medial amygdaloid nucleus may be a pivotal player in acute responses to emotional environmental stimuli. METHODS: The present study therefore aimed to study the effects of bilateral electrolytic lesions of the medial amygdala on unconditioned anxiety-related behavior as well as a sensorimotor gating parameter (prepulse inhibition, PPI) in rats. Anxiety-related behavior was assessed with the use of stress-induced hyperthermia (SIH), light-enhanced startle (LES) and open field behavior. RESULTS: Bilateral electrolytic lesions of the medial amygdala decreased the SIH response and anxiety-related open field behavior. In contrast, lesioned animals displayed augmented LES and disrupted PPI. No changes in basal locomotor activity, body temperature and acoustic startle were found between lesioned and sham animals. CONCLUSIONS: The present study suggests that the medial amygdala is an important player in response to acute environmental stimuli. Decreased unconditioned psychological stress responses were found, whereas LES was enhanced and sensorimotor processing was disrupted. However, considering the existing data on basolateral amygdala involvement in PPI and bed nucleus of the stria terminalis involvement in LES, local infusion studies into the MeA should be performed to further substantiate these findings. PMID- 20006966 TI - Region-specific alteration in brain glutamate: possible relationship to risk taking behavior. AB - Risk-taking behaviors involve increased motor activity and reduced anxiety in humans. Total sleep deprivation (SD) in animals produces a similar change in motor and fear behaviors. Investigators studied region-specific brain levels of glutamate in rats after TSD, an animal model of risk-taking behavior. We investigated the effects of sleep deprivation on these behaviors and associated levels of brain glutamate. Compared to the controls, the sleep-deprived rats spent a significantly greater percentage of time in the open arms of the elevated plus maze (EPM), demonstrating reduced fear-like and increased risk-taking behaviors. Additionally, sleep deprivation was associated with a significant increase in glutamate levels in the hippocampus and thalamus. An inverse relationship between glutamate in the medial prefrontal cortex and risk taking in the EPM and a positive association between the ratio of glutamate in the hippocampus to medial prefrontal cortex and risk taking was revealed. The role of sleep deprivation-induced changes in brain glutamate and its relationship to anxiety, fear, and posttraumatic stress disorder (PTSD) is discussed. PMID- 20006967 TI - Opioid receptor antagonism in the nucleus accumbens fails to block the expression of sugar-conditioned flavor preferences in rats. AB - In our prior studies, systemic administration of the opioid receptor antagonist naltrexone (NTX) did not block flavor preference conditioning by the sweet taste or post-oral actions of sugar despite reducing intake. Because opioid signaling in the nucleus accumbens (NAc) is implicated in food reward, this study determined if NTX administered into the NAc would block the expression of sugar conditioned preferences. In Experiment 1, food-restricted rats with bilateral NAc shell or core cannulae were trained to drink a fructose (8%)+saccharin (0.2%) solution mixed with one flavor (CS+) and a less-preferred 0.2% saccharin solution mixed with another flavor (CS-) during one-bottle sessions. Two-bottle tests with the two flavors mixed in saccharin solutions occurred 10 min following total bilateral NAc shell or core doses of 0, 1, 25 and 50 microg of NTX. The rats preferred the CS+ over CS- following vehicle (80%) and all NTX doses in the shell and core. The CS+ preference was reduced to 64% and 72% by 50 microg NTX in the shell and core, although only the core effect was significant. In Experiment 2, food-restricted rats were trained to drink one flavored saccharin solution (CS+) paired with an intragastic (IG) glucose (8%) infusion and a second flavored saccharin solution (CS-) paired with an IG water infusion. In subsequent two bottle tests, the rats displayed significant preferences for the CS+ (81-91%) that were unaltered by any NTX dose in the shell or core. CS+ intake, however, was reduced by NTX in the shell, but not the core. These data indicate that accumbal opioid antagonism slightly attenuated, but did not block the expression of sugar-conditioned flavor preferences. Therefore, while opioid drugs can have potent effects on sugar intake they appear less effective in altering sugar conditioned flavor preferences. PMID- 20006968 TI - Individual differences in activity predict locomotor activity and conditioned place preference to amphetamine in both adolescent and adult rats. AB - Individual and developmental differences in novelty seeking have been implicated in differential sensitivity to psychostimulants in rodents, but findings are mixed. The extent to which age differences in activity in a novel arena depended on test duration was examined by comparing adolescent and adult rats after 5 and after 60 min of testing (session 1). Rats were tested again after amphetamine or saline administration 24h later (session 2) to examine whether activity in a novel arena predicts sensitivity to locomotor-activating effects of amphetamine. Data from two experiments were used to examine consistency of the findings. Only activity in 60 min sessions produced a consistent age difference (adolescentpH 5.0) evoked [Ca(2+)](i) increases, but not in DRG neurons from transient receptor potential V1 (TRPV1) ( /-) mice. Regardless of isolectin GS-IB4 (IB4)-staining, histamine potentiated [Ca(2+)](i) responses to acid (>or=pH 6.0) that were mediated by TRPV1 activation. Histamine increased membrane depolarization induced by acid and evoked spike discharges. RT-PCR indicated the expression of all four histamine receptors (H1R, H2R, H3R, H4R) in mouse DRG. The potentiating effect of histamine was mimicked by an H1R agonist, but not H2R-H4R agonists and was inhibited only by an H1R antagonist. Histamine failed to potentiate the [Ca(2+)](i) response to acid in the presence of inhibitors for phospholipase C (PLC) and protein kinase C (PKC). A lipoxygenase inhibitor and protein kinase A inhibitor did not affect the potentiating effects of histamine. Carrageenan and complete Freund's adjuvant produced inflammatory hyperalgesia, but these inflammatory conditions did not change the potentiating effects of histamine in DRG neurons. The present results suggest that histamine sensitizes acid-induced responses through TRPV1 activation via H1R coupled with PLC/PKC pathways, the action of which may be involved in the generation of inflammatory pain. PMID- 20006974 TI - 5-HT1A receptors mediate detrimental effects of cocaine on long-term potentiation and expression of polysialylated neural cell adhesion molecule protein in rat dentate gyrus. AB - The present study investigated the involvement of 5-HT(1A) receptors in the inhibitory effect of single administration of cocaine (COC, 15 mg/kg i.p.) on the induction of long-term potentiation (LTP) in slices of rat dentate gyrus (DG), prepared 30 min and 2 days after COC administration. These effects of COC were blocked by an antagonist of 5-HT(1A) receptors, WAY 100635 (0.4 mg/kg i.p.), which had been administered 20 min before COC. The detrimental effect of COC on LTP in slices prepared 30 min after COC administration could be prevented by blocking glucocorticoid receptors (GRs) using mifepristone (RU 38486, 10 mg/kg s.c. given 1 h before COC), similar as in slices obtained 2 days after COC as reported previously [Mackowiak et al. (2008) Eur J Neurosci 27:2928-2937]. After a single administration of an agonist of 5-HT(1A) receptors, 8-OH-DPAT, (0.5 mg/kg i.p.), the level of LTP in slices prepared 2 days later was significantly decreased resembling the effect of COC. This effect of 8-OH-DPAT was antagonized by WAY 100635 (0.4 mg/kg i.p.), administered 20 min before 8-OH-DPAT and by RU 38486, given 1 h before 8-OH-DPAT. COC-induced inhibition of LTP could be blocked by the inhibitor of mitogen-activated protein kinase kinase 1/2 (MEK1/2), SL 327 (50 mg/kg i.p.), administered 1 h before COC, but not by the inhibitor of phosphatidylinositol 3-kinase (PI3-kinase), LY 294002 (80 mg/kg i.p.). COC induced reduction in the number of polysialylated neural cell adhesion molecule (PSA-NCAM)-positive neurons in rat dentate gyrus could also be prevented by WAY 100635, given 20 min before COC. These data indicate that the indirect 5-HT(1A) receptor activation by a single COC administration and subsequent stimulation of extracellular signal-regulated kinases (ERK 1/2) signaling pathway result in a decrease of the potential for long-term increase in synaptic efficacy in rat DG lasting at least two but less than 7 days, most likely via activation of the hypothalamic-pituitary-adrenal (HPA) axis. PMID- 20006975 TI - Seizures are associated with brain injury severity in a neonatal model of hypoxia ischemia. AB - Hypoxia-ischemia is a significant cause of brain damage in the human newborn and can result in long-term neurodevelopmental disability. The loss of oxygen and glucose supply to the developing brain leads to excitotoxic neuronal cell damage and death; such over-excitation of nerve cells can also manifest as seizures. The newborn brain is highly susceptible to seizures although it is unclear what role they have in hypoxic-ischemic (H/I) injury. The aim of this study was to determine an association between seizures and severity of brain injury in a piglet model of perinatal H/I and, whether injury severity was related to type of seizure, i.e. sub-clinical (electrographic seizures only) or clinical (electrographic seizures+physical signs). Hypoxia (4% O(2)) was induced in anaesthetised newborn piglets for 30 min with a final 10 min period of hypotension; animals were recovered and survived to 72 h. Animals were monitored daily for seizures both visually and with electroencephalogram (EEG) recordings. Brain injury was assessed with magnetic resonance imaging (MRI), (1)H-MR spectroscopy ((1)H-MRS), EEG and by histology (haematoxylin and eosin). EEG seizures were observed in 75% of all H/I animals, 46% displayed clinical seizures and 29% sub-clinical seizures. Seizure animals showed significantly lower background amplitude EEG across all post-insult days. Presence of seizures was associated with lower cortical apparent diffusion coefficient (ADC) scores and changes in (1)H-MRS metabolite ratios at both 24 and 72 h post-insult. On post mortem examination animals with seizures showed the greatest degree of neuropathological injury compared to animals without seizures. Furthermore, clinical seizure animals had significantly greater histological injury compared with sub-clinical seizure animals; this difference was not apparent on MRI or (1)H-MRS measures. In conclusion we report that both sub-clinical and clinical seizures are associated with increased severity of H/I injury in a term model of neonatal H/I. PMID- 20006976 TI - Antiviral flavonoids from Mosla scabra. AB - An ethanolic extract and ethyl acetate fraction (EFMS) prepared from Mosla scabra showed potential inhibition against influenza viruses. The compounds isolated from EFMS were elucidated to be ursolic acid (1), 5-hydroxy-6,7-dimethoxyflavone (2), 5-hydroxy-7,8-dimethoxyflavone (3), apigenin (4) and acacetin (5) based on their spectral analysis. Compounds 2, 3, 4 and 5 showed significant anti influenza viruses activities. Compared with ribavirin, EFMS appeared to have similar efficacy against influenza viruses, was considered to be less toxic and may warrant further evaluation as a possible agent for the treatment of influenza. PMID- 20006977 TI - Sesquiterpenoids and phenolics from roots of Taraxacum udum. AB - From roots of Taraxacum udum, two new and four known sesquiterpene lactones were isolated, together with five known phenolic compounds. The new compounds were characterized as 11beta, 13-dihydrotaraxinic acid and taraxinic acid 6-O-acetyl beta-glucopyranosyl ester by spectroscopic methods, especially 1D and 2D NMR, and by comparison with structurally related compounds. The plant material was shown to be a good source of taraxinic acid derivatives. PMID- 20006978 TI - An overview of foodborne pathogen detection: in the perspective of biosensors. AB - Food safety is a global health goal and the foodborne diseases take a major crisis on health. Therefore, detection of microbial pathogens in food is the solution to the prevention and recognition of problems related to health and safety. For this reason, a comprehensive literature survey has been carried out aiming to give an overview in the field of foodborne pathogen detection. Conventional and standard bacterial detection methods such as culture and colony counting methods, immunology-based methods and polymerase chain reaction based methods, may take up to several hours or even a few days to yield an answer. Obviously this is inadequate, and recently many researchers are focusing towards the progress of rapid methods. Although new technologies like biosensors show potential approaches, further research and development is essential before biosensors become a real and reliable choice. New bio-molecular techniques for food pathogen detection are being developed to improve the biosensor characteristics such as sensitivity and selectivity, also which is rapid, reliable, effective and suitable for in situ analysis. This paper not only offers an overview in the area of microbial pathogen detection but it also describes the conventional methods, analytical techniques and recent developments in food pathogen detection, identification and quantification, with an emphasis on biosensors. PMID- 20006979 TI - Elucidating the transcriptome of Fasciola hepatica - a key to fundamental and biotechnological discoveries for a neglected parasite. AB - Liver flukes of animals are parasitic flatworms (Platyhelminthes: Digenea) of major socioeconomic importance in many countries. Key representatives, such as Fasciola hepatica and F. gigantica, cause "liver fluke disease" (= fascioliasis), which is of major animal health significance worldwide. In particular, F. hepatica is a leading cause of production losses to the livestock (mainly sheep and cattle) and meat industries due to clinical disease, reduced weight gain and milk production, and deaths. This parasite is also a major food-borne pathogen of humans throughout parts of the Middle East, Asia and South America. Currently, there is a significant focus on the development of new approaches for the prevention and control of fascioliasis in livestock. Recent technological advances in genomics and bioinformatics provide unique opportunities for the identification and prevalidation of drug targets and vaccines through a better understanding of the biology of F. hepatica and related species as well as their relationship with their hosts at the molecular level. Surprisingly, despite the widespread socioeconomic impact of fascioliasis, genomic datasets for F. hepatica are scant, limiting the molecular biological research of this parasite. The present article explores specifically the transcriptome of the adult stage of F. hepatica using an integrated genomic-bioinformatic platform. The analysis of the current data reveals numerous molecules of biological relevance, some of which are inferred to be involved in key biological processes or pathways that could serve as targets for new trematocidal drugs or vaccines. Improved insights into the transcriptome of F. hepatica should pave the way for future, comparative analysis of the transcriptomes of other developmental stages of this and related parasites, such as F. gigantica, cancer-causing flatworms (Clonorchis sinensis and Opisthorchis viverrini) and blood flukes (Schistosoma mansoni and S. japonicum). Prediction of the essentiality of genes and their products, molecular network connectivity of trematode genes as well as experimental exploration of function should also add value to the genomic discovery efforts in the future, focused on biotechnological outcomes. PMID- 20006980 TI - Okadaic acid induces apoptosis in Down syndrome fibroblasts. AB - Down's syndrome (DS) is characterized by several pathological aspects leading to an increased susceptibility to cardiovascular diseases, infections, leukemia, endocrine alterations. DS patients display some of the physiopathological characteristics of aging, observed also in Alzheimer disease (AD), such as abnormalities in lipids metabolism, diabetes, high cholesterol fraction, senile plaques and neurofibrillary tangles. For this reason DS is considered a precocious and accelerated model of senescence, in which increased apoptosis is the main cornerstone. In order to better understand the apoptotic process in pathological cellular aspects of DS, the aim of this study was to investigate the apoptotic response of DS fibroblasts to OA, a toxin that induces malformations and inhibits growth in different cell lines. We focused specifically on the mitochondrial response by investigating changes in mitochondrial membrane potential (evaluate by flow cytometry and fluorescence microscopy using JC-1 probe) and alterations of mitochondrial outer membrane (evaluated by flow cytometry using annexin V/propidium iodide). Results indicates that DS Fibroblasts have a baseline of apoptosis higher than normal fibroblasts and are more susceptible to the pro-apoptotic effect of OA. Understanding the mechanism of apoptosis in DS fibroblasts could provide new insight in the pathogenic mechanism of this pathology and suggest potential therapeutical targets to the clinical treatment at complex diseases associated to this pathology. PMID- 20006981 TI - Regulation of VDR expression in rat and human intestine and liver--consequences for CYP3A expression. AB - The vitamin D receptor (VDR) regulates the expression of drug metabolizing enzymes and transporters in intestine and liver, but the regulation of VDR expression in intestine and liver is incompletely understood. We studied the regulation of VDR mRNA expression by ligands for VDR, farnesoid X receptor (FXR), glucocorticoid receptor (GR) and protein kinase C alpha (PKCalpha) in rat and human ileum and liver using precision-cut slices. 1,25(OH)(2)D(3) induced VDR expression in rat ileum and liver, and human ileum but not in liver. Chenodeoxycholic acid (CDCA), but not lithocholic acid (LCA) and GW4064 induced VDR mRNA expression in rat ileum and liver. The PKCalpha activator, phorbol-12 myristate-13-acetate (PMA) induced the expression of VDR in the rat liver, and the induction of VDR by 1,25(OH)(2)D(3) and CDCA was inhibited by the PKCalpha inhibitor, bisindolyl maleimide I (Bis I). These results show that the expression of VDR is likely to be regulated by PKC but not by FXR or VDR activation at least in the rat liver. The VDR mediated induction of its target genes CYP3A1 and CYP3A2 by 1,25(OH)(2)D(3) or LCA in the rat ileum was strongly reduced in the presence of CDCA despite the higher VDR expression. Thus, CDCA might potentiate the toxicity of LCA by inhibiting its metabolism. PMID- 20006982 TI - Dispersion characteristics of various metal oxide secondary nanoparticles in culture medium for in vitro toxicology assessment. AB - The aim of this study is to characterize the dispersion characteristics of various metal oxide nanoparticles and secondary nanoparticle formation in culture medium. Many studies have already investigated the in vitro toxicities of various metal oxide nanoparticles; however, there have been few discussions about the particle transport mode to cells during a period of toxicity assessment. The particle transport mode would strongly affect the amount of uptake by cells; therefore, estimation of the transport mode for various metal oxide particles is important. Fourteen different metal oxide nanoparticle dispersions in a culture medium were examined. The sizes of the secondary nanoparticles were observed to be larger than 100 nm by dynamic light scattering (DLS). According to Stokes law and the Stokes-Einstein assumption, pure metal oxide particles with such sizes should gravitationally settle faster than diffusion processes; however, the secondary metal oxide particles examined in this study exhibited unexpectedly slower gravitational settling rates. The slow gravitational settling kinetics of particles was estimated to be caused by the inclusion of protein into the secondary nanoparticles, which resulted in lower densities than the pure metal oxide particles. The ratios of metal oxide to protein in secondary particles could be affected by the protein adsorption ability of the corresponding metal oxide primary particles. To the best of our knowledge, it was clarified for the first time that stably dispersed secondary metal oxide nanoparticles with slow gravitational settling kinetics are induced by secondary nanoparticles consisting of small amounts of metal oxide particles and large amounts of protein, which results in lower particle densities than the pure metal oxide particles. The estimation of particle dynamics in culture medium using this method would be significant to recognize the inherent toxicity of nanoparticles. PMID- 20006983 TI - Dishevelled: The hub of Wnt signaling. AB - Wnt signaling controls a variety of developmental and homeostatic events. As a key component of Wnt signaling, Dishevelled (Dvl/Dsh) protein relays Wnt signals from receptors to downstream effectors. In the canonical Wnt pathway that depends on the nuclear translocation of beta-catenin, Dvl is recruited by the receptor Frizzled and prevents the constitutive destruction of cytosolic beta-catenin. In the non-canonical Wnt pathways such as Wnt-Frizzled/PCP (planar cell polarity) signaling, Dvl signals via the Daam1-RhoA axis and the Rac1 axis. In addition, Dvl plays important roles in Wnt-GSK3beta-microtubule signaling, Wnt-calcium signaling, Wnt-RYK signaling, Wnt-atypical PKC signaling, etc. Dvl also functions to mediate receptor endocytosis. To fulfill its multifaceted functions, it is not surprising that Dvl associates with various kinds of proteins. Its activity is also modulated dynamically by phosphorylation, ubiquitination and degradation. In this review, we summarize the current understanding of Dvl functions in Wnt signal transduction and its biological functions in mouse development, and also discuss the molecular mechanisms of its actions. PMID- 20006984 TI - Characterization of coproporphyrinogen III oxidase in Plasmodium falciparum cytosol. AB - A unique hybrid pathway has been proposed for de novo heme biosynthesis in Plasmodium falciparum involving three different compartments of the parasite, namely mitochondrion, apicoplast and cytosol. While parasite mitochondrion and apicoplast have been shown to harbor key enzymes of the pathway, there has been no experimental evidence for the involvement of parasite cytosol in heme biosynthesis. In this study, a recombinant P. falciparum coproporphyrinogen III oxidase (rPfCPO) was produced in E. coli and confirmed to be active under aerobic conditions. rPfCPO behaved as a monomer of 61kDa molecular mass in gel filtration analysis. Immunofluorescence studies using antibodies to rPfCPO suggested that the enzyme was present in the parasite cytosol. These results were confirmed by detection of enzyme activity only in the parasite soluble fraction. Western blot analysis with anti-rPfCPO antibodies also revealed a 58kDa protein only in this fraction and not in the membrane fraction. The cytosolic presence of PfCPO provides evidence for a hybrid heme-biosynthetic pathway in the malarial parasite. PMID- 20006985 TI - Expanding school psychology's horizons for understanding and conducting research: commentary on the special series on statistical analysis. PMID- 20006986 TI - An introduction to modern missing data analyses. AB - A great deal of recent methodological research has focused on two modern missing data analysis methods: maximum likelihood and multiple imputation. These approaches are advantageous to traditional techniques (e.g. deletion and mean imputation techniques) because they require less stringent assumptions and mitigate the pitfalls of traditional techniques. This article explains the theoretical underpinnings of missing data analyses, gives an overview of traditional missing data techniques, and provides accessible descriptions of maximum likelihood and multiple imputation. In particular, this article focuses on maximum likelihood estimation and presents two analysis examples from the Longitudinal Study of American Youth data. One of these examples includes a description of the use of auxiliary variables. Finally, the paper illustrates ways that researchers can use intentional, or planned, missing data to enhance their research designs. PMID- 20006987 TI - Curriculum-based measurement of math problem solving: a methodology and rationale for establishing equivalence of scores. AB - The purpose of this article is to discuss curriculum-based measurement (CBM) as it is currently utilized in research and practice and to propose a new approach for developing measures to monitor the academic progress of students longitudinally. To accomplish this, we first describe CBM and provide several exemplars of CBM in reading and mathematics. Then, we present the research context for developing a set of seven curriculum-based measures for monitoring student progress in math problem solving. The rationale for and advantages of using statistical equating methodology are discussed. Details of the methodology as it was applied to the development of these math problem solving measures are provided. PMID- 20006988 TI - Evaluating mediation and moderation effects in school psychology: a presentation of methods and review of current practice. AB - Third variable effects elucidate the relation between two other variables, and can describe why they are related or under what conditions they are related. This article demonstrates methods to analyze two third-variable effects: moderation and mediation. The utility of examining moderation and mediation effects in school psychology is described and current use of the analyses in applied school psychology research is reviewed and evaluated. Proper statistical methods to test the effects are presented, and different effect size measures for the models are provided. Extensions of the basic moderator and mediator models are also described. PMID- 20006989 TI - A practical guide to multilevel modeling. AB - Collecting data from students within classrooms or schools, and collecting data from students on multiple occasions over time, are two common sampling methods used in educational research that often require multilevel modeling (MLM) data analysis techniques to avoid Type-1 errors. The purpose of this article is to clarify the seven major steps involved in a multilevel analysis: (1) clarifying the research question, (2) choosing the appropriate parameter estimator, (3) assessing the need for MLM, (4) building the level-1 model, (5) building the level-2 model, (6) multilevel effect size reporting, and (7) likelihood ratio model testing. The seven steps are illustrated with both a cross-sectional and a longitudinal MLM example from the National Educational Longitudinal Study (NELS) dataset. The goal of this article is to assist applied researchers in conducting and interpreting multilevel analyses and to offer recommendations to guide the reporting of MLM analysis results. PMID- 20006990 TI - Chiral silica-based monoliths in chromatography and capillary electrochromatography. AB - Chiral-modified silica-based monoliths have become well-established stationary phases for both high performance liquid chromatography (HPLC) and capillary electrochromatography (CEC). The silica-based monoliths were fabricated either in situ in the capillaries for nano-HPLC and CEC or in a mould for "conventional" HPLC. The present review summarizes the chiral modification of silica monoliths and the recent development in the field of enantioselective separations by nano HPLC and CEC. PMID- 20006991 TI - Peptide-based spherulitic films--formation and properties. AB - Peptide nanotube-based spherulitic films are a recently discovered phenomenon, which was demonstrated in the case of the self-assembled diphenylalanine peptide nanotubes. Here we show that the film-formation method can be implemented with other peptides. We also demonstrate that a critical physical parameter, an elevated level of environmental hydration, is required for film growth. A possible formation mechanism is suggested. The optical, morphological and mechanical properties of these films are characterized and are found to be substantially different from those of non-spherulitic deposits. PMID- 20006992 TI - Correlation of a set of gene variants, life events and personality features on adult ADHD severity. AB - Increasing evidence suggests that symptoms of attention deficit hyperactivity disorder (ADHD) could persist into adult life in a substantial proportion of cases. The aim of the present study was to investigate the impact of (1) adverse events, (2) personality traits and (3) genetic variants chosen on the basis of previous findings and (4) their possible interactions on adult ADHD severity. One hundred and ten individuals diagnosed with adult ADHD were evaluated for occurrence of adverse events in childhood and adulthood, and personality traits by the Temperament and Character Inventory (TCI). Common polymorphisms within a set of nine important candidate genes (SLC6A3, DBH, DRD4, DRD5, HTR2A, CHRNA7, BDNF, PRKG1 and TAAR9) were genotyped for each subject. Life events, personality traits and genetic variations were analyzed in relationship to severity of current symptoms, according to the Brown Attention Deficit Disorder Scale (BADDS). Genetic variations were not significantly associated with severity of ADHD symptoms. Life stressors displayed only a minor effect as compared to personality traits. Indeed, symptoms' severity was significantly correlated with the temperamental trait of Harm avoidance and the character trait of Self directedness. The results of the present work are in line with previous evidence of a significant correlation between some personality traits and adult ADHD. However, several limitations such as the small sample size and the exclusion of patients with other severe comorbid psychiatric disorders could have influenced the significance of present findings. PMID- 20006994 TI - Identification of two auto-cleavage products of nonstructural protein 1 (nsp1) in porcine reproductive and respiratory syndrome virus infected cells: nsp1 function as interferon antagonist. AB - The porcine reproductive and respiratory syndrome virus nsp1 is predicted to be auto-cleaved from the replicase polyprotein into nsp1alpha and nsp1beta subunits. In infected cells, we detected the actual existence of nsp1alpha and nsp1beta. Cleavage sites between nsp1alpha/nsp1beta and nsp1beta/nsp2 were identified by protein microsequencing analysis. Time course study showed that nsp1alpha and nsp1beta mainly localize into the cell nucleus after 10 h post infection. Further analysis revealed that both proteins dramatically inhibited IFN-beta expression. The nsp1beta was observed to significantly inhibit expression from an interferon stimulated response element promoter after Sendai virus infection or interferon treatment. It was further determined to inhibit nuclear translocation of STAT1 in the JAK-STAT signaling pathway. These results demonstrated that nsp1beta has ability to inhibit both interferon synthesis and signaling, while nsp1alpha alone strongly inhibits interferon synthesis. These findings provide important insights into mechanisms of nsp1 in PRRSV pathogenesis and its impact in vaccine development. PMID- 20006993 TI - A tale of tails: Sialidase is key to success in a model of phage therapy against K1-capsulated Escherichia coli. AB - Prior studies treating mice infected with Escherichia coli O18:K1:H7 observed that phages requiring the K1 capsule for infection (K1-dep) were superior to capsule-independent (K1-ind) phages. We show that three K1-ind phages all have low fitness when grown on cells in serum whereas fitnesses of four K1-dep phages were high. The difference is serum-specific, as fitnesses in broth overlapped. Sialidase activity was associated with all K1-dep virions tested but no K1-ind virions, a phenotype supported by sequence analyses. Adding endosialidase to cells infected with K1-ind phage increased fitness in serum by enhancing productive infection after adsorption. We propose that virion sialidase activity is the primary determinant of high fitness on cells grown in serum, and thus in a mammalian host. Although the benefit of sialidase is specific to K1-capsulated bacteria, this study may provide a scientific rationale for selecting phages for therapeutic use in many systemic infections. PMID- 20006995 TI - Inverse relationships between selenium and mercury in tissues of young walleye (Stizosedion vitreum) from Canadian boreal lakes. AB - The concentrations of total mercury (Hg), methylmercury (MeHg) and total selenium (Se) were determined in muscle, liver and brain tissues of young-of-the-year walleye (Stizosedion vitreum) specimens collected from 8 boreal lakes that are located within 107km around the Sudbury smelters in Ontario, Canada. Dry weight basis concentrations of Hg were highest in muscle and lowest in brain (p<0.05), those of MeHg were higher in muscle than in liver and brain but there was no significant difference between liver and brain (p<0.05). The highest Se concentrations were found in liver and the lowest in brain (p<0.05). Considering the biomass of the studied tissues, muscle was the part of the body where most of Hg, MeHg and Se were accumulated. In fish muscle, the percentage of MeHg over Hg was the highest and this percentage was the lowest in liver. The concentrations of Hg, MeHg and Se in the studied tissues were closely related to the concentrations of total dissolved Se in lake waters which vary with the distance of the lakes from the smelters. Thresholds of Se concentrations in tissues were revealed (6.2, 12.0 and 3.5mgkg(-1) dry wt., for muscle, liver and brain, respectively), above which a significant reduction of MeHg concentrations was observed in all studied tissues compared to lower Se levels in the same tissues. Based on the collected information and data analysis, possible mechanisms for the biological processes behind the observed inverse relationships between Se and Hg in fish tissues are discussed. PMID- 20006996 TI - Recombinant coagulation factor VIIa--from molecular to clinical aspects of a versatile haemostatic agent. AB - Recombinant factor VIIa (rFVIIa, NovoSeven) is currently the only bypassing agent produced by recombinant technology for the treatment of haemophiliacs whose disease is complicated by inhibitory antibodies. In addition, recombinant production of FVIIa has made it widely available for a variety of purposes and accelerated the growth of our knowledge about FVIIa by generating an abundance of clinical and biochemical data. This fascinating molecule has turned out to be a safe haemostatic agent with great potential in the clinic and has inspired the generation of improved variants currently in (pre-)clinical testing. The present review describes the structural and functional aspects of FVIIa, followed by sections dealing with the manufacturing, therapeutic mechanism of action, clinical development and experience with rFVIIa. PMID- 20006997 TI - OSU-03012 sensitizes TIB-196 myeloma cells to imatinib mesylate via AMP-activated protein kinase and STAT3 pathways. AB - Although c-Kit is expressed on the surface of myeloma cells in one-third of myeloma patients, the efficacy of imatinib mesylate for patients with myeloma is still controversial. To investigate the combinatorial effect of OSU-03012 and imatinib mesylate, we treated a c-Kit-expressing myeloma cell line, TIB-196, with DMSO, OSU-03012 alone, imatinib mesylate alone and OSU-03012 plus imatinib mesylate. OSU-03012 sensitized TIB-196 cells to imatinib mesylate cytotoxicity. p STAT3 (Tyr705), as well as down-stream cyclin D1 and Mcl-1, was down regulated. Additionally, there was markedly increased p-AMPK (Thr172) and down-regulation of p-p70S6K (Thr386) in the combination group. Combined treatments targeting c-Kit, AMPK and STAT3 may be a potential strategy for treating patients with myeloma. PMID- 20006998 TI - DNA methylation does not regulate JUNB expression in CML: comment on "Downregulation of JUNB mRNA expression in advanced phase chronic myelogenous leukemia" by Hoshino et al. [Leuk. Res. 33 (2009) 1361-1366]. PMID- 20006999 TI - How does a valine residue that modulates heme-NO binding kinetics in inducible NO synthase regulate enzyme catalysis? AB - Nitric oxide (NO) release from nitric oxide synthases (NOSs) depends on the dissociation of a ferric heme-NO product complex (Fe(III)NO) that forms immediately after NO is made in the heme pocket. The NOS-like enzyme of Bacillus subtilis (bsNOS) has 10-20 fold slower Fe(III)NO dissociation rate (k(d)) and NO association rate (k(on)) compared to mammalian NOS counterparts. We previously showed that an Ile for Val substitution at the opening of the heme pocket in bsNOS contributes to these differences. The complementary mutation in mouse inducible NOS oxygenase domain (Val346Ile) decreased the NO k(on) and k(d) by 8 and 3-fold, respectively, compared to wild-type iNOSoxy, and also slowed the reductive processing of the heme-O(2) catalytic intermediate. To investigate how these changes affect steady-state catalytic behaviors, we generated and characterized the V346I mutant of full-length inducible NOS (iNOS). The mutant exhibited a 4-5 fold lower NO synthesis activity, an apparent uncoupled NADPH consumption, and formation of a heme-NO complex during catalysis that was no longer sensitive to solution NO scavenging. We found that these altered catalytic behaviors were not due to changes in the heme reduction rate or in the stability of the enzyme heme-O(2) intermediate, but instead were due to the slower NO k(on) and k(d) and a slower oxidation rate of the enzyme ferrous heme-NO complex. Computer simulations that utilized the measured kinetic values confirmed this interpretation, and revealed that the V346I iNOS has an enhanced NADPH-dependent NO dioxygenase activity that converts almost 1 NO to nitrate for every NO that the enzyme releases into solution. Together, our results highlight the importance of heme pocket geometry in tuning the NO release versus NO dioxygenase activities of iNOS. PMID- 20007000 TI - Non-covalent interactions in blue copper protein probed by Met16 mutation and electronic and resonance Raman spectroscopy of Achromobacter cycloclastes pseudoazurin. AB - We have used low-temperature (77K) resonance Raman (RR) spectroscopy as a probe of the electronic and molecular structure to investigate weak pi-pi interactions between the metal ion-coordinated His imidazoles and aromatic side chains in the second coordination sphere of blue copper proteins. For this purpose, the RR spectra of Met16 mutants of Achromobacter cycloclastes pseudoazurin (AcPAz) with aromatic (Met16Tyr, Met16Trp, and Met16Phe) and aliphatic (Met16Ala, Met16Val, Met16Leu, and Met16Ile) amino acid side chains have been obtained and analyzed over the 100-500cm(-1) spectral region. Subtle strengthening of the Cu(II)-S(Cys) interaction on replacing Met16 with Tyr, Trp, and Phe is indicated by the upshifted (0.3-0.8cm(-1)) RR bands involving nu(Cu-S)(Cys) stretching modes. In contrast, the RR spectra of Met16 mutants with aliphatic amino acids revealed larger (0.2-1.8cm(-1)) shifts of the nu(Cu-S)(Cys) stretching modes to a lower frequency region, which indicate a weakening of the Cu(II)-S(Cys) bond. Comparisons of the predominantly nu(Cu-S)(Cys) stretching RR peaks of the Met16X=Tyr, Trp, and Phe variants, with the molar absorptivity ratio epsilon(1)/epsilon(2) of sigma( approximately 455nm)/pi( approximately 595nm) (Cys)S-->Cu(II) charge-transfer bands in the optical spectrum and the axial/rhombic EPR signals, revealed a slightly more trigonal disposition of ligands about the copper(II) ion. In contrast, the RR spectra of Met16Z=Ala, Val, Leu, and Ile variants with aliphatic amino acid side chains show a more tetrahedral perturbation of the copper active site, as judged by the lower frequencies of the nu(Cu-S)(Cys) stretching modes, much larger values of the epsilon(1)/epsilon(2) ratio, and the increased rhombicity of the EPR spectra. PMID- 20007002 TI - Unsuspected pulmonary emboli in lung cancer patients: the impact on survival and the significance of anticoagulation therapy. AB - BACKGROUND: Many pulmonary emboli (PE) are detected unsuspectedly in lung cancer patients. The purpose of our study was to retrospectively evaluate the role of anticoagulation therapy for unsuspected PE in lung cancer patients. We also aimed to evaluate risk factors associated with the development of PE as well as the prognostic power of PE in lung cancer patients. PATIENTS AND METHODS: The Samsung Medical Information System was used to evaluate predictors and prognosis of PE in lung cancer patients. We found patients with PE using the Radiation Interpretation Registry and reviewed their medical records. RESULTS: Among 8014 lung cancer patients, PE developed in 180 patients (cumulative incidence rates=2.2%). Metastasis and prior history of chemotherapy were significant predictors of the development of PE. Pulmonary embolism detected within 3 months after diagnosis of lung cancer was a significant poor prognostic factor (hazard ratio [HR], 1.5; 95% CI, 1.1-2.0) in the complete lung cancer cohort. One hundred thirteen (63%) out of total 180 PE patients were incidentally found to have PE. Among the 113 patients with unsuspected PE, 62 patients (55%) did not receive anticoagulation therapy, and died sooner than those who received anticoagulation therapy for unsuspected PE (HR, 4.1; 95% CI, 2.3-7.6). CONCLUSION: Anticoagulation therapy for unsuspected PE is associated with increased overall survival in lung cancer patients. PMID- 20007001 TI - Reaction of the zinc sensor FluoZin-3 with Zn(7)-metallothionein: Inquiry into the existence of a proposed weak binding site. AB - It has been reported that Zn(7)-metallothionein (MT), contains one weak binding site for Zn(2+). To test this conclusion, rabbit liver MT isolated at pH 7 was reacted with chelating agents of modest affinity for Zn(2+). Contrary to the previous study, no evidence was found for Zn(2+) stoichiometrically bound to the protein with an apparent stability constant of about 10(8). Indeed, stability constant measurements based upon competition between Zn(7)-MT and ligands of known stability with Zn(2+) showed that all of the protein bound Zn(2+) displayed the same stability constant at pH 7.4 and 25 degrees C of (1.7+/-0.6)x10(11). Brief reaction of Zn(7)-MT with strong acid converted it into MT(*) and upon reneutralization into Zn(7)-MT(*), which demonstrated reactivity of about 1 Zn(2+)/mol MT with competing ligands. Acid titration of Zn(7)-MT to pH 2 or below rapidly resulted in the formation of Zn(7)-MT(*) that displayed biphasic titration with base, revealing the rebinding of lower affinity Zn(2+) between pH 5 and 7. Since MT is commonly acidified during preparation, care must be taken to document which form of the protein is present in subsequent experiments at pH 7. PMID- 20007003 TI - Pain in patients with lung cancer: pathophysiology and treatment. AB - This review analyses the characteristics of the principal pain syndromes associated with lung cancer, their physiopathology and causes, and provides updated information on available treatments. Pain associated with lung cancer is characterized by multiple expressions, due to either the progression of disease and/or induced by oncological treatment. The analgesic treatment is principally based on the use of opioids. Other than the oral route, which is the preferred one, alternative modalities to administer opioids may be helpful in different clinical circumstances. According to the opioid response, other routes and other opioids, may improve the balance between analgesia and adverse effects providing the best individual response to a specific opioid drug. More complex strategies, such as interventional procedures, are seldom necessary and require an appropriate selection of patients. PMID- 20007004 TI - Stability of osteosynthesis with bicortical screws placed in a triangular shape in mandibular sagittal split 5 mm advancement osteotomy: biomechanical tests. AB - Sagittal split ramus osteotomy (SSRO) is common in orthognathic surgery, and osteosynthesis with bicortical titanium screws placed in a triangular shape is used for stabilisation. We biomechanically tested the stability of this configuration in 5 mm advancement SSRO with condylar replacement in the initial position using six fresh frozen human mandibles raised subperiosteally for surgical simulation. Osteotomy was done after marking the initial positions of the condyles and the symphysis on graph paper. The condyle was set in its initial position and the symphysis advanced exactly 5mm; this position was maintained during osteosynthesis using 2.7 mm bicortical screws placed in a triangular shape. The mandible was then placed horizontally on the testing machine and a continuous static force was applied perpendicularly at a displacement speed of 5 mm/min. A stainless steel plate was used to transmit the forces independently of the teeth. A mean force of 129.6 Newtons (N) was applied at the elastic/plastic limit on an effort/displacement curve, which is well beyond the maximal mean (SD) chewing forces of 21 N (14) measured after bilateral sagittal split ramus osteotomy (BSSRO). A linear equation relating force to displacement was worked out from this experiment. Osteosynthesis using three bicortical screws mounted triangularly after advancement SSRO of 5 mm is experimentally stable, and our results could be used experimentally to compare two osteosyntheses using the equation. PMID- 20007005 TI - Cardiac arrest in the catheterisation laboratory: a 5-year experience of using mechanical chest compressions to facilitate PCI during prolonged resuscitation efforts. AB - PURPOSE: Lengthy resuscitations in the catheterisation laboratory carry extremely high rates of mortality because it is essentially impossible to perform effective chest compressions during percutaneous coronary intervention (PCI). The purpose of this study was to evaluate the use of a mechanical chest compression device, LUCAS, in the catheterisation laboratory, in patients who suffered circulatory arrest requiring prolonged resuscitation. MATERIALS AND METHODS: The study population was comprised of patients who arrived alive to the catheterisation laboratory and then required mechanical chest compression at some time during the angiogram, PCI or pericardiocentesis between 2004 and 2008 at the Lund University Hospital. This is a retrospective registry analysis. RESULTS: During the study period, a total of 3058 patients were treated with PCI for ST-elevation myocardial infarction (STEMI) of whom 118 were in cardiogenic shock and 81 required defibrillations. LUCAS was used in 43 patients (33 STEMI, 7 non-ST elevation myocardial infarction (NSTEMI), 2 elective PCIs and 1 patient with tamponade). Five patients had tamponade due to myocardial rupture prior to PCI that was revealed at the start of the PCI, and all five died. Of the remaining 38 patients, 1 patient underwent a successful pericardiocentesis and 36 were treated with PCI. Eleven of these patients were discharged alive in good neurological condition. CONCLUSION: The use of mechanical chest compressions in the catheterisation laboratory allows for continued PCI or pericardiocentesis despite ongoing cardiac or circulatory arrest with artificially sustained circulation. It is unlikely that few, if any, of the patients would have survived without the use of mechanical chest compressions in the catheterisation laboratory. PMID- 20007006 TI - Equilibrium and kinetic studies of adsorption of Cd(II) from aqueous solution using modified corn stalk. AB - This paper describes the adsorption of cadmium ions from aqueous solution using acrylonitrile (AN)-modified corn stalk (AMCS). AMCS was characterized by elemental analysis, scanning electron microscopy, surface area and porosity analyzer, Fourier transform infrared and solid-state CP/MAS (13)C NMR spectra, and then used to evaluate the adsorption capacity in different pH values, adsorption isotherm, kinetics and thermodynamics in batch experiments. The results showed that AMCS is found to be an effective adsorbent because of its pore size and functional groups (-CN). The pH of 7.0 was an optimal pH for removal of Cd(II) ion and the Langmuir model provides a better fit to the equilibrium data than the Freundlich model, showing a maximum uptake of 12.73 mg g(-1), compared to raw corn stalk (RCS) (3.39 mg g(-1)). Analysis indicated that pseudo-second-order kinetics controlled the adsorption rate. The activation energy (E(a)) was 9.43 kJ mol(-1). Thermodynamic parameters such as DeltaG, DeltaH and DeltaS were also evaluated to predict the nature of adsorption process. PMID- 20007007 TI - Facing-target sputtering deposition of ZnO films with Pt ultra-thin layers for gas-phase photocatalytic application. AB - In this paper, various zinc oxide (ZnO) films are deposited by a versatile and effective dc-reactive facing-target sputtering method. The ratios of Ar to O(2) in the mixture gas are varied from 8:2 to 6:4 at a fixed sputtering pressure of 1.0 Pa. X-ray diffraction, spectrophotometer and scanning electron microscope are used to study the crystal structure, optical property and surface morphology of the as-deposited films. The Pt ultra-thin layer, approximately 2 nm thick, is deposited on the surface of ZnO film by dc diode sputtering with a mesh mask controlling the coated area. The photocatalytic activity of ZnO films and Pt-ZnO films is evaluated by decomposition of methanol under UV-vis light irradiation. The variation of photocatalytic activity depends on the ratios of Ar to O(2), which is mainly attributed to the different grain size and carrier mobility. Though the pure ZnO film normally shows a low gas-phase photocatalytic activity, its activity is significantly enhanced by depositing Pt ultra-thin layer. PMID- 20007008 TI - Preparation of ZnO/Cu2O compound photocatalyst and application in treating organic dyes. AB - ZnO/Cu(2)O compound photocatalysts were prepared by "soak-deoxidize-air oxidation" with different concentrations of Cu(2+) (0.125, 0.25, 0.5, 1, 1.5 and 2 mol/L). The prepared ZnO/Cu(2)O samples were characterized by X-ray diffraction (XRD), scanning electron microscopy (SEM), energy-dispersive spectrometer (EDS), UV-vis diffuse reflectance spectrometer, and photochemical reaction instrument. The results show that ZnO was hexagonal wurtzite structure and the crystallinity had no change with the increase of Cu(2+) concentration. Cu(2)O belonged to cubic structure and the crystallinity increased with the increase of Cu(2+) concentration. ZnO were rods and bulks which had diameter of about 300-400 nm, some small round Cu(2)O particles which had a diameter of about 50 nm adhered to these rods and bulks. In the compounds the mole ratio of Cu(2)O to ZnO was 0.017, 0.025, 0.076, 0.137, 0.138, and 0.136, respectively. An absorbance in the visible light region between 400 and 610 nm was seen and the reflection rate became less with the mole ratio of Cu(2)O to ZnO increasing. The photocatalytic activities of ZnO/Cu(2)O compound were evaluated using a basic organic dye, methyl orange (MO). It was found that, compared with pure ZnO, the photocatalytic properties of ZnO/Cu(2)O compound were improved greatly and some compounds were better than pure Cu(2)O. PMID- 20007009 TI - A procedure for the assessment of the toxicity of intermediates and products formed during the accidental thermal decomposition of a chemical species. AB - The knowledge of the substances which form when a molecule undergoes chemical reactions under unusual conditions is required by European legislation to evaluate the risks associated with an industrial chemical process. A thermal decomposition is often the result of a loss of control of the process which leads to the formation of many substances in some cases not easily predictable. The evaluation of the change of an overall toxicity passing from the parent compound to the mixture of its thermal decomposition products has been already proposed as a practical approach to this problem when preliminary indications about the temperature range in which the molecule decomposes are available. A new procedure is proposed in this work for the obtainment of the mixtures of thermal decomposition products also when there is no previous information about the thermal behaviour of investigated molecules. A scanning calorimetric run that is aimed to identify the onset temperature of the decomposition process is coupled to an isoperibolic one in order to obtain and collect the products. An algal strain is adopted for toxicological assessments of chemical compounds and mixtures. An extension of toxicological investigations to human cells is also attempted. PMID- 20007010 TI - Acetazolamide and midazolam act synergistically to inhibit neuropathic pain. AB - Treatment of neuropathic pain is a major clinical challenge that has been met with minimal success. After peripheral nerve injury, a decrease in the expression of the K-Cl cotransporter KCC2, a major neuronal Cl(-) extruder, leads to pathologic alterations in GABA(A) and glycine receptor function in the spinal cord. The down-regulation of KCC2 is expected to cause a reduction in Cl(-) extrusion capacity in dorsal horn neurons, which, together with the depolarizing efflux of HCO(3)(-) anions via GABA(A) channels, would result in a decrease in the efficacy of GABA(A)-mediated inhibition. Carbonic anhydrases (CA) facilitate intracellular HCO(3)(-) generation and hence, we hypothesized that inhibition of CAs would enhance the efficacy of GABAergic inhibition in the context of neuropathic pain. Despite the decrease in KCC2 expression, spinal administration of benzodiazepines has been shown to be anti-allodynic in neuropathic conditions. Thus, we also hypothesized that spinal inhibition of CAs might enhance the anti allodynic effects of spinally administered benzodiazepines. Here, we show that inhibition of spinal CA activity with acetazolamide (ACT) reduces neuropathic allodynia. Moreover, we demonstrate that spinal co-administration of ACT and midazolam (MZL) act synergistically to reduce neuropathic allodynia after peripheral nerve injury. These findings indicate that the combined use of CA inhibitors and benzodiazepines may be effective in the clinical management of neuropathic pain in humans. PMID- 20007011 TI - Dydrogesterone support in threatened miscarriage. AB - INTRODUCTION: The aim of this study was to determine whether dydrogesterone helps to preserve pregnancy in women with threatened miscarriage. METHODS: 146 women who presented with mild or moderate vaginal bleeding during the first trimester of pregnancy were randomised to receive oral dydrogesterone (10mg b.i.d.) (n=86) or no treatment (n=60). Dydrogesterone was continued until 1 week after the bleeding had stopped. All women received standard supportive care. RESULTS: The incidence of miscarriage was significantly lower in the dydrogesterone group than in the untreated group (17.5% vs. 25%; p<0.05). There were no statistically significant differences between the groups with respect to pregnancy complications or congenital abnormalities. CONCLUSION: Dydrogesterone appears to have beneficial effects in women with threatened miscarriage. PMID- 20007012 TI - Low-dose multidetector-row CT-angiography of abdominal aortic aneurysm after endovascular repair. AB - PURPOSE: To investigate the possibility of reducing radiation dose exposure while maintaining image quality using multidetector computed tomography angiography (MDCTA) with high-concentration contrast media in patients undergoing follow-up after endovascular aortic repair (EVAR) to treat abdominal aortic aneurysm. MATERIALS AND METHODS: In this prospective, single center, intra-individual study, patients underwent two consecutive MDCTA scans 6 months apart, one with a standard acquisition protocol (130 mAs/120 kV) and 120 mL of iomeprol 300, and one using a low dose protocol (100 mAs/80 kV) and 90 mL of iomeprol 400. Images acquired during the arterial phase of contrast enhancement were evaluated both qualitatively and quantitatively for image noise and intraluminal contrast enhancement. RESULTS: Thirty adult patients were prospectively enrolled. Statistically significantly higher attenuation values were measured in the low dose acquisition protocol compared to the standard protocol, from the suprarenal abdominal aorta to the common femoral artery (p<0.0001; all vascular segments). Qualitatively, image quality was judged significantly (p=0.0002) better with the standard protocol than with the low-dose protocol. However, no significant differences were found between the two protocols in terms of contrast-to-noise ratio (CNR) (13.63+/-6.97 vs. 11.48+/-8.13; p=0.1058). An overall dose reduction of up to 74% was observed for the low-dose protocol compared with the standard protocol. CONCLUSION: In repeat follow-up examinations of patients undergoing EVAR for abdominal aortic aneurysm, a low-dose radiation exposure acquisition protocol provides substantially reduced radiation exposure while maintaining a constant CNR and good image quality. PMID- 20007013 TI - LC-NMR identification of a novel taurine-related metabolite observed in (1)H NMR based metabonomics of genetically hypertensive rats. AB - This paper describes the LC-NMR spectroscopic identification of a novel urinary endogenous metabolite responsible for the signals, which were found as major contributors to the separation between genetically hypertensive rats (SHRSP) and normotensive control rats (WKY) in previous NMR-based metabonomic studies. Urine samples from 26-week-old normotensive rats were analyzed by an LC-NMR system equipped with a reversed-phase column having high retention ability for polar compounds. (1)H NMR spectra were continuously obtained in the on-flow mode, and the retention times of the unassigned signals in question were determined. Various two-dimensional spectra were subsequently measured for the fraction containing the unassigned signals under the stop-flow mode, which enables for a long accumulation resulting in the enhancement of signal-to-noise ratios. The candidate compound obtained from these LC-NMR data was synthesized, and the NMR and mass spectra were compared with those of the LC-NMR fraction. The unknown metabolite was identified as succinyltaurine from these experiments together with standard addition experiments. This novel metabolite, which is characteristic of the normotensive rats, is very interesting because it is structurally related to hypotensive taurine, and not substantially detected in the genetically hypertensive rats, which excreted more taurine than the normotensive rats. The biological and pathophysiological significances of succinyltaurine remain to be investigated. PMID- 20007014 TI - Electrochemical determination of NADH and ethanol based on ionic liquid functionalized graphene. AB - It is firstly reported that low-potential NADH detection and biosensing for ethanol are achieved at an ionic liquid-functionalized graphene (IL-graphene) modified electrode. A substantial decrease (440 mV) in the overvoltage of the NADH oxidation was observed using IL-graphene/chitosan coating, with oxidation starting at ca. 0 V (vs. Ag|AgCl). And the NADH amperometric response at such a modified electrode is more stable (95.4% and 90% of the initial activity remaining after 10 min and 30 min at 1 mM NADH solution) than that at bare electrode (68% and 46%). Furthermore, the IL-graphene/chitosan-modified electrode exhibited a good linearity from 0.25 to 2 mM and high sensitivity of 37.43 microA mM(-1)cm(-2). The ability of IL-graphene to promote the electron transfer between NADH and the electrode exhibited a novel and promising biocompatible platform for development of dehydrogenase-based amperometric biosensors. With alcohol dehydrogenase (ADH) as a model, the ADH/IL-graphene/chitosan-modified electrode was constructed through a simple casting method. The resulting biosensor showed rapid and highly sensitive amperometric response to ethanol with a low detection limit (5 microM). Moreover, the proposed biosensor has been used to determine ethanol in real samples and the results were in good agreement with those certified by the supplier. PMID- 20007015 TI - Clinical pharmacokinetics of the new oral camptothecin gimatecan: the inter patient variability is related to alpha1-acid glycoprotein plasma levels. AB - AIM OF THE STUDY: To determine the pharmacokinetics of gimatecan, a camptothecin with a lipophilic substitution in position 7, given orally to patients participating in the phase I study. METHODS: Pharmacokinetics was evaluated in 78 patients after oral daily dose for 5 days a week for 1, 2 or 3 weeks by HPLC with a fluorescence detector. RESULTS: Gimatecan was mainly present in plasma as lactone (>85%), the active form as DNA-topoisomerase I poison. The AUC(0-24) on the first day of treatment normalised per daily dose (mg/m(2)), ranged from 194 to 2909 ng h/mL/mg/m(2). The half-life was 77.1+/-29.6h, consequently C(max) and AUC rose 3-6-fold after multiple dosing. Multivariate analysis indicated the daily dose (p<0.0001) and the alpha(1)-acid glycoprotein (AGP) plasma levels (p<0.0001) as main predictors of gimatecan AUC(0-24). In the overall analysis, daily dose and AGP plasma levels explained 85% of the deviance. The hydroxy metabolite ST1698 was present in plasma at low levels with AUC values of 5-15% of gimatecan. In mice, orally treated with gimatecan, plasma and tissue levels were 2-fold higher after treatment with a pro-inflammatory agent causing AGP induction. CONCLUSIONS: Gimatecan is orally absorbed and its variable plasma levels seem to be related to AGP plasma concentrations. Data obtained in mice, together with the fact that AGP levels largely exceeded gimatecan plasma concentrations, suggest that the increased gimatecan levels in patients with high AGP levels are not related to the binding of the drug to AGP with consequent reduced tissue drug distribution, but possibly to other mechanism associated with inflammation being AGP simply a marker of the inflammation process. PMID- 20007016 TI - Lumber quality of Eucalyptus grandis as a function of diametrical position and log steaming. AB - The objective of this study was to evaluate the effect of log steaming and of the diametrical position of boards on the timber quality of Eucalyptus grandis. Logs with diameters between 20 and 25 cm, between 25 and 30 cm and between 30 and 35 cm were studied. Half of logs were kept in its original condition, and the other half was steamed at 90 degrees C for 20 h. Later, the logs were cut into flat saw boards, and defects due to growth stress relief were measured. The results show that: (1) boards from control logs show different magnitudes of cracking according to the diameter of the log and the diametrical position of the board; (2) boards from logs with diameters between 30 and 35 cm and those from next to the pith develop larger cracks; and (3) boards from steamed logs show a reduction in the magnitude of cracking and a homogenous distribution of this defect relative to diametrical position within the log. PMID- 20007017 TI - Synthesis and biological evaluation of substituted 2-phenyl-2H-indazole-7 carboxamides as potent poly(ADP-ribose) polymerase (PARP) inhibitors. AB - A potent series of substituted 2-phenyl-2H-indazole-7-carboxamides were synthesized and evaluated as inhibitors of poly (ADP-ribose) polymerase (PARP). This extensive SAR exploration culminated with the identification of substituted 5-fluoro-2-phenyl-2H-indazole-7-carboxamide analog 48 which displayed excellent PARP enzyme inhibition with IC(50)=4nM, inhibited proliferation of cancer cell lines deficient in BRCA-1 with CC(50)=42nM and showed encouraging pharmacokinetic properties in rats compared to the lead 6. PMID- 20007018 TI - Knowledge applied to new domains: the unconscious succeeds where the conscious fails. AB - A common view holds that consciousness is needed for knowledge acquired in one domain to be applied in a novel domain. We present evidence for the opposite; where the transfer of knowledge is achieved only in the absence of conscious awareness. Knowledge of artificial grammars was examined where training and testing occurred in different vocabularies or modalities. In all conditions grammaticality judgments attributed to random selection showed above-chance accuracy (60%), while those attributed to conscious decisions did not. Participants also rated each string's familiarity and performed a perceptual task assessing fluency. Familiarity was predicted by repetition structure and was thus related to grammaticality. Fluency, though increasing familiarity, was unrelated to grammaticality. While familiarity predicted all judgments only those attributed to random selection showed a significant additional contribution of grammaticality, deriving primarily from chunk novelty. In knowledge transfer, as in visual perception (Marcel, 1993), the unconscious may outperform the conscious. PMID- 20007020 TI - Wavelet optimization for content-based image retrieval in medical databases. AB - We propose in this article a content-based image retrieval (CBIR) method for diagnosis aid in medical fields. In the proposed system, images are indexed in a generic fashion, without extracting domain-specific features: a signature is built for each image from its wavelet transform. These image signatures characterize the distribution of wavelet coefficients in each subband of the decomposition. A distance measure is then defined to compare two image signatures and thus retrieve the most similar images in a database when a query image is submitted by a physician. To retrieve relevant images from a medical database, the signatures and the distance measure must be related to the medical interpretation of images. As a consequence, we introduce several degrees of freedom in the system so that it can be tuned to any pathology and image modality. In particular, we propose to adapt the wavelet basis, within the lifting scheme framework, and to use a custom decomposition scheme. Weights are also introduced between subbands. All these parameters are tuned by an optimization procedure, using the medical grading of each image in the database to define a performance measure. The system is assessed on two medical image databases: one for diabetic retinopathy follow up and one for screening mammography, as well as a general purpose database. Results are promising: a mean precision of 56.50%, 70.91% and 96.10% is achieved for these three databases, when five images are returned by the system. PMID- 20007019 TI - Variable Field Proton-Electron Double-Resonance Imaging: Application to pH mapping of aqueous samples. AB - A new concept of Variable Field Proton-Electron Double-Resonance Imaging (VF PEDRI) is proposed. This allows for functional mapping using specifically designed paramagnetic probes (e.g. oxygen or pH mapping) with MRI high quality spatial resolution and short acquisition time. Studies performed at 200 G field MRI with phantoms show that a pH map of the sample can be extracted using only two PEDRI images acquired in 140 s at pre-selected EPR excitation fields providing pH resolution of 0.1 pH units and a spatial resolution of 1.25mm. Note that while concept of functional VF PEDRI was demonstrated using the pH probe, it can be applied for studies of other biologically relevant parameters of the medium such as redox state, concentrations of oxygen or glutathione using specifically designed EPR probes. PMID- 20007021 TI - Photophysical properties of the 8-phenyl analogue of PM567: a theoretical rationalization. AB - The absorption and emission spectroscopic investigations of the C-8-phenyl substituted analogue of the pyrromethene dye PM567 in various polar, non-polar as well as protic and aprotic solvents are reported. The solvatochromic shifts of the spectral bands were studied in a multitude of polar, non-polar and protic, aprotic solvents followed by a multilinear regression in which several solvent parameters were simultaneously analysed. Comparison of the experimental results with those obtained by gas phase ab initio computation with CIS, TD-HF and TD-DFT theories using 6-31G* basis set reveal an overestimation of the experimentally measured excitation parameters by all these theoretical models. However, the trends in the experimental results agree with those calculated theoretically. PMID- 20007022 TI - Competition between solvent quenching and indole quenching of 9-fluorenone: a spectroscopic and computational study. AB - The interaction between 9-fluorenone, various indoles and solvents has been studied using steady-state fluorescence spectroscopy and quantum chemical calculations. It was determined that polar protic solvents such as methanol and ethanol significantly quenched the fluorescence of 9-fluorenone but various indoles reversed the solvent quenching. The effect of various solvents on the 9 fluorenone carbonyl vibration was investigated using infrared spectroscopy. Ab initio calculations using Gaussian03 were also carried out in order to determine the minimum energy conformations of these systems along with binding energies. PMID- 20007023 TI - Enteroviruses in Spain over the decade 1998-2007: virological and epidemiological studies. AB - BACKGROUND: Human enteroviruses (HEV) are the commonest cause of viral meningitis as well as other pathologies, therefore HEV characterization is important both in patient management and epidemiological investigation. OBJECTIVES: A 10-year study of patients with enteroviral infection was carried out in Spain to determine the underlying etiology. STUDY DESIGN: HEV were fully typed by microneutralisation tests and/or molecular methods. RESULTS: A collection of 86404 clinical samples were studied in several Spanish laboratories. These were collected from patients with different syndromes, mainly aseptic meningitis (AM), fever, respiratory diseases and acute flaccid paralysis. Of these, 6867 HEV were obtained. At the National Poliovirus Laboratory 2814 were serotypically characterised. Among non polio enteroviruses, the eight main serotypes were Echovirus 30 (25%), Echovirus 6 (12.4%), Echovirus 13 (8.3%), Echovirus 11 (7.4%) and Echovirus 9 (4.7%), followed by Coxsackievirus B5 (4.2%) and Echovirus 7 and Coxsackievirus A9 (3.7%) each. In AM cases, Echovirus 30 was identified in 39% of them, followed by Echovirus 6 (14%). However, Echovirus 6 was mainly associated with respiratory disease (17%), followed by Echovirus 11 (10%). On the other hand, Echovirus 30, Echovirus 11 and Echovirus 6 contributed equally with 12% of each serotype in the cases of fever. CONCLUSIONS: The present report complements previous data (Trallero et al.(13)), with the results of HEV incidence in Spain from 1998 to 2007. The surveillance described in this study provided valuable information as to which serotypes are in circulation, the emergence of new HEV and association with clinical manifestations. PMID- 20007024 TI - Swine infuenza (H1N1) pneumonia: bacterial airway colonization common but fatalities due to bacterial pneumonia remain relatively rare. PMID- 20007025 TI - Exponential stability on stochastic neural networks with discrete interval and distributed delays. AB - This brief addresses the stability analysis problem for stochastic neural networks (SNNs) with discrete interval and distributed time-varying delays. The interval time-varying delay is assumed to satisfy 0 < d(1) 0.05); in the common bile duct cyst group, the EK positive rate was significantly lower than the trypsin-1 positive rate (P<0.05). CONCLUSIONS: Western blotting can accurately reflect duodenal-biliary and pancreaticobiliary refluxes. EK has greater sensitivity than RN for duodenal-biliary reflux. The majority of biliary amylase and lipase comes from the pancreas in all biliary diseases; pancreaticobiliary reflux is the predominant source in the common bile duct cyst group and duodenal-biliary reflux is responsible for the ductal pigment stone group. PMID- 20007079 TI - Actions of genistein on contractile response of smooth muscle isolated from guinea pig gallbladder. AB - BACKGROUND: Defective contractile motility of the gallbladder is an important factor for gallstone formation. Estrogen might increase the risk of gallstones and cholecystitis, and estradiol inhibits the contractile activity of isolated strips of guinea pig gallbladder. The potential risks associated with hormone replacement therapy (HRT) include symptomatic gallstones. Phytoestrogen have been used to treat menopause syndromes by replacing traditional estrogen. This experiment aimed to determine the effects of the phytoestrogen genistein on the contractile response of smooth muscle strips isolated from guinea pig gallbladder and its possible mechanism of action. METHODS: Guinea pigs were sacrificed to remove the whole gallbladder. Two or three smooth muscle strips were cut longitudinally. Each strip was suspended in a tissue chamber containing Krebs solution. After 2 hours of equilibration, contractile response indexes were recorded. Different concentrations of genistein were added to the chamber and the contractile responses were measured. Each antagonist was added 2 minutes before genistein to study possible mechanisms. The effect of genistein on calcium dependent contraction curves and biphasic contraction in calcium-free Krebs solution were measured. RESULTS: Genistein decreased the resting tension dose dependently, and reduced the mean contractile amplitude and frequency in gallbladder strips. Ranitidine partly inhibited the effect of genistein, but methylene blue, Nomega-nitro-L-arginine, and propranolol hydrochloride did not influence this action. Genistein had no significant effects on calcium-dependent contraction. Genistein reduced the first contraction induced by acetylcholine chloride, but did not affect the second contraction caused by CaCl2. CONCLUSIONS: Genistein relaxed smooth muscle isolated from the gallbladder of guinea pigs and this might contribute to the formation of gallstones. The inhibitory action might be related to H2 receptors and the release of intracellular Ca2+ from sarcoplasmic reticulum. Replacing traditional estrogen with phytoestrogen to treat menopause syndromes may increase the risk of gallstone formation. PMID- 20007080 TI - CA19-9 antigen levels can distinguish between benign and malignant pancreaticobiliary disease. AB - BACKGROUND: CA19-9 is a carbohydrate tumor-associated antigen which is frequently upregulated in pancreatobiliary neoplasia. However, it may also be elevated in patients with jaundice in the absence of a tumor due to biliary obstruction, and in other non-hepato-pancreatico-biliary conditions. This study aimed to evaluate whether CA19-9 levels could accurately differentiate between benign and malignant pancreatobiliary disease. METHODS: All patients referred to a single surgeon for investigation of pancreaticobiliary disease in 2003 in whom a firm diagnosis had been established were included. For malignant disease, a histological diagnosis was required but for benign disease a firm radiological diagnosis was deemed adequate. The patients were divided into 4 categories: pancreatic adenocarcinoma (PCa); cholangiocarcinoma (CCa); chronic pancreatitis (CP) and biliary calculous disease (Calc). Bilirubin and alkaline phosphatase levels corresponding to the point of assessment of CA19-9 were also noted. RESULTS: Final diagnoses were made of pancreatic adenocarcinoma (PCa, n=73), cholangiocarcinoma (CCa, n=19), ampullary carcinoma (Amp, n=7), neuroendocrine carcinoma (Neu, n=4), duodenal carcinoma (Duo, n=3), chronic pancreatitis (CP, n=115), and biliary calculous disease (Calc, n=27). Median CA19-9 levels (U/ml) were: PCa, 653; CCa, 408; Duo, 403; Calc, 27; CP, 19; Neu, 10.5; Amp, 8 (reference range: 0-37). The CA19-9 levels were significantly greater for malignant than for benign disease, could differentiate PCa from CCa/Duo, and were significantly higher in unresectable than in resectable PCa. The sensitivity, specificity, positive predictive value (PPV) and negative predictive value (NPV) for CA19-9 were 84.9%, 69.7%, 67.7% and 86.1%, respectively. A ROC analysis provided an area under the curve for CA19-9 of 0.871 (0.820-0.922), giving an optimal CA19-9 of 70.5 U/ml for differentiating benign from malignant pathology. Using this cut-off, the sensitivity was 82.1%, while specificity, PPV and NPV improved to 85.9%, 81.3% and 86.5%, respectively. When standard radiology was included (US/CT/MRCP) in the decision process, the results improved to 97.2%, 88.7%, 86.6%, and 97.7%. For benign disease, the CA19 9 correlated directly with the serum bilirubin, but for malignant disease, CA19-9 levels were elevated independent of the bilirubin level. CONCLUSIONS: CA19-9 is useful in the differentiation of pancreatobiliary disease and when using an optimized cut-off and combining with routine radiology, the diagnostic yield is improved significantly, thus stressing the importance of a multi-disciplinary approach to pancreatobiliary disease. PMID- 20007081 TI - Pathogenic role of myeloperoxidase in acute pancreatitis. AB - BACKGROUND: Myeloperoxidase (MPO) has been implicated in promoting tissue damage in various inflammatory diseases. However, MPO blood levels in relation to the severity of acute pancreatitis (AP) and its time-course have not been studied. The present study aimed to determine the role of MPO in AP. METHODS: We studied 86 patients with AP (48 patients with mild and 38 with severe pancreatitis) and 18 controls (volunteers). The relations of serum MPO levels to cytokine level, severity, and time-course of pancreatitis were studied. The serum level of MPO and cytokines were measured by MPO-EIA and cytokines ELISA, respectively. RESULTS: The highest level of MPO was noted at the first day in patients with severe AP. A decrease of MPO blood level occurred during the first three days in all patients with necrotizing pancreatitis. The development of pancreatitis associated lung injury and purulent complications was accompanied by increased MPO levels. Administration of pentoxifylline significantly reduced the MPO blood level, which was clearly correlated with the levels of proinflammatory cytokines in the two groups of patients. CONCLUSIONS: The results of the present study showed the MPO blood level is dependent on the severity of AP and on cytokine blood levels. Pentoxifylline in the complex management of severe AP may improve the results of treatment. PMID- 20007082 TI - Cholangiopancreatography troubleshooting: the usefulness of endoscopic retrieval of migrated biliary and pancreatic stents. AB - BACKGROUND: Stent migration in the hepatopancreatic duct might arise as one of the rare complications associated with biliary or pancreatic stenting. Although there are some procedures to retrieve the migrated stent, including surgical, percutaneous, and endoscopic approaches, endoscopy should be attempted first because it is least invasive. This study set out to evaluate the usefulness of endoscopic retrieval of migrated biliary and pancreatic stents. METHODS: Plastic stents that migrated in the bile duct (35 patients) or pancreatic duct (2) were retrieved with endoscopic retrograde cholangiopancreatography. Devices used were snare forceps, a basket catheter, grasping forceps, biopsy forceps, a balloon catheter, and the Soehendra stent retriever. RESULTS: Endoscopic retrieval of migrated stents was performed successfully in 36 (97.0%) of the 37 patients. The devices utilized for successful treatment were basket catheter (13 patients), grasping forceps (10), snare forceps (8), balloon catheter (3), biopsy forceps (1), and the Soehendra stent retriever (1). The unsuccessfully treated patient with chronic pancreatitis underwent surgery since the guide wire did not move forward due to bile duct stenosis, and there was also duodenal stenosis. One patient developed mild pancreatitis after withdrawal of the stent; the pancreatitis was relieved with conservative treatment. CONCLUSIONS: Endoscopic retrieval of migrated biliary and pancreatic stents appears to be useful because of its safety and low invasiveness. However, various forceps should be prepared for the retrieval of a migrated stent. PMID- 20007083 TI - Ruptured hydatid cyst in a patient with shock. AB - Echinococcosis is a severe helminthic zoonosis largely caused by Echinococcus granulosus and frequently encountered in endemic areas. The liver and lung are the most frequently involved organs. Cyst rupture into the peritoneal cavity represents a rare but serious complication. Herein, we describe an acute occurrence of anaphylactic shock due to a spontaneous rupture of hydatid cyst in a 21-year-old Turkish patient unwittingly infected by Echinococcosis. Resection surgery of the perforated cyst in combination with cleaning of the abdominal cavity was performed. The patient rapidly improved and no relapse occurred during a follow-up of 8 months. Anaphylaxis is a serious complication of hydatid cyst rupture and needs to be promptly diagnosed. The main objective of our report is to underscore this life-threatening complication that should be considered when anaphylactic shock of unknown origin occurs, even in non-endemic regions. Moreover, we emphasize the need for a radical surgical approach to avoid widespread dissemination. PMID- 20007085 TI - Choledochal cyst with large stone cast and portal hypertension. AB - BACKGROUND: Choledochal cysts in adults are more commonly associated with complications such as cystolithiasis, recurrent cholangitis, portal hypertension and malignancy, than in the pediatric age group. METHOD: We report a case of adult choledochal cyst with long-term complication of large stone cast and portal hypertension due to secondary biliary cirrhosis. RESULTS: A 50-year-old patient presented with obstructive jaundice and hepatosplenomegaly. On investigation, she was diagnosed as having a choledochal cyst with large stone cast and portal hypertension. Single stage resection of the choledochal cyst with Roux-en-Y hepaticojejunostomy was done by meticulous dissection and ligation of collaterals. CONCLUSION: Single stage resection of a choledochal cyst is possible in spite of associated portal hypertension, if the portal vein is patent. PMID- 20007084 TI - Malignant carcinoid tumor of the cystic duct: a rare cause of bile duct obstruction. AB - BACKGROUND: Carcinoid tumors of the extrahepatic biliary tree are extremely rare malignancies, accounting for 0.2%-2% of all gastrointestinal carcinoid tumors, while carcinoids of the cystic duct are an uncommon entity and an extremely unusual cause of bile duct obstruction. METHODS: After an extensive literature review, we retrospectively analysed 61 cases of carcinoid tumor of the biliary tree as well as one additional case of a 60-year-old female with symptoms and laboratory/imaging examination findings compatible with those of a malignant biliary tract obstruction. At laparotomy, resection of the gallbladder and common bile duct was performed. Histological study revealed a well-differentiated neuro endocrine carcinoma of the cystic duct. The patient remained disease-free at 16 months. RESULTS: Our presentation is the seventh case reported in the world literature. Compared to cholangiocarcinoma, analysis of the reviewed group indicates an increased incidence of extrahepatic carcinoid tumors in younger persons along with a slight female predominance. Statistically, the most common anatomic location is the common bile duct, followed by the perihilar region and the cystic duct. Jaundice is the most common finding. Curative surgery was realized in the majority of cases and long-term disease-free survival was achieved when surgery was curative. CONCLUSIONS: Carcinoid tumors obstructing the biliary tree are extremely difficult to diagnose preoperatively, and nearly impossible to differentiate from non-neuroendocrine tumors. As surgery offers the only potential cure for both biliary carcinoids and cholangiocarcinoma, we recommend aggressive surgical therapy as the treatment of choice in every case of potentially resectable biliary tumor. PMID- 20007086 TI - Extrahepatic intraductal ectopic hepatocellular carcinoma: bile duct filling defect. AB - BACKGROUND: Obstructive jaundice caused by an intraductal hepatocellular carcinoma is a rare initial symptom. We report a rare case of an extrahepatic icteric type hepatocellular carcinoma. METHODS: A 75-year-old patient was admitted to our hospital because of obstructive jaundice 3 months after resection of multilocular hepatocellular carcinoma. A postoperative bile leakage was treated by placement of a decompressing stent in the common bile duct. Endoscopic retrograde choledochoscopy showed extended blood clots filling the bile duct system and computed tomography revealed a local swelling in the common extrahepatic bile duct. The level of alpha-fetoprotein (AFP) was only slightly elevated but that of CA19-9 was dramatically increased. Cholangiography showed an intraductal filling defect typical of a cholangiocellular carcinoma. RESULTS: Bile duct brushing cytology showed no cholangiocellular carcinoma but hepatocellular carcinoma cells in the extrahepatic bile duct. An extrahepatic bile duct resection was performed. Histological examination confirmed the diagnosis of extrahepatic intraductal growth of hepatocellular carcinoma. CONCLUSION: Ectopic hepatocellular carcinoma is a rare but important differentially diagnosed of extrahepatic bile duct filling defect. PMID- 20007087 TI - Autoimmune pancreatitis: report of two cases and literature review. AB - BACKGROUND: Autoimmune pancreatitis (AIP) is a unique chronic inflammation of the pancreas in which autoimmune mechanisms are involved in the pathogenesis. It is characterized by clinical, histopathological, radiographic, serologic and therapeutic features. Since it was first described in 1995, increasing numbers of AIP patients have been diagnosed. METHODS: In this study, the data from 2 patients with AIP who had elevation of serum immunoglobulin G, positive autoantibody, swelling of the pancreas and narrowing of the main pancreatic duct on imaging were analyzed retrospectively. RESULTS: With the initial diagnosis of AIP, both patients underwent regular prednisone therapy, which was initiated at a dose of 40 mg per day with a tapering schedule of 5 mg every 2 weeks. The patients responded very well to treatment with prednisone. CONCLUSIONS: AIP is a relatively new disease entity. Although it is diagnosed by distinct characteristics of the clinical, radiologic, histopathologic, and serologic findings, many questions require further clarification, including its relationship to other autoimmune diseases and misdiagnosis as pancreaticobiliary malignancies. PMID- 20007088 TI - Subacute fulminant hepatic failure with intermittent fever. AB - BACKGROUND: Viral hepatitis B accounts for over 80% of acute hepatic failures in China and the patients die mainly of its complications. A patient with hepatic failure and fever is not uncommon, whereas repeated fever is rare. METHODS: A 32 year-old female was diagnosed with subacute hepatic failure and hepatitis B viral infection because of hyperbilirubinemia, coagulopathy, hepatic encephalopathy, serum anti-HBs-positive without hepatitis B vaccination, and typical intrahepatic pathological features of chronic hepatitis B. Plasma exchange was administered twice and she awoke with hyperbilirubinemia and discontinuous fever. RESULTS: Urethritis was confirmed and medication-induced fever and/or spontaneous bacterial peritonitis (Gram-negative bacillus infection) was suspected. The patient was treated with antibiotics, steroids and a Chinese herbal medicine, matrine, for three months and she recovered. CONCLUSION: The survival rate of patients with hepatic failure might be improved with comprehensive supporting measures and appropriate, timely management of complications. PMID- 20007105 TI - Chronic myeloid leukemia: where do we go now? PMID- 20007106 TI - The chronic myeloid leukemia stem cell. AB - Chronic myeloid leukemia (CML) is a clonal stem cell disorder that is characterized by the acquired chromosomal translocation BCR-ABL. This gives rise to a constitutively active tyrosine kinase deregulation of the normal mechanisms of cell cycle control. In the normal hematopoietic system, hematopoietic stem cells (HSC) self-renew to form identical daughter cells but also differentiate to mature blood cells. Leukemic stem cells (LSC) share these properties of self renewal and also differentiate to mature leukemic cells. LSC have been isolated from patients with CML: these cells give rise to leukemia following transplantation into NOD-SCID mice models. Further characterization of CML stem cells has demonstrated that a small percentage of these cells are quiescent despite culture with growth factors. The CML stem cell arises from a normal HSC that has acquired the Philadelphia chromosome. In advanced phase, more mature cells such as granulocyte/monocyte progenitors might also acquire the ability to self-renew and function as LSC. This might be one of the mechanisms underlying the progression to blast crisis. Quiescent stem cells are resistant to treatment with imatinib in vitro and are thought also to show resistance in vivo. The properties of the stem cells that lead to this drug resistance are still being characterized. However, this drug insensitivity leads to disease persistence that may lead to disease relapse even despite an initial response to imatinib. Newer molecular therapies are in development that act to specifically target and eradicate the stem cell pool. PMID- 20007107 TI - Standard management of patients with chronic myeloid leukemia. AB - The successful introduction of the tyrosine kinase inhibitors (TKIs) has revolutionized the treatment of patients with chronic myeloid leukemia (CML). Imatinib therapy induces high rates of complete cytogenetic and major molecular responses, and improves survival in CML. Following imatinib treatment, more than 90% of patients obtain complete hematologic response, and over 80% achieve a complete cytogenetic response. With 7 years of follow-up, the results are still very favorable, resulting in a major change in the natural history of the disease. Resistance to imatinib represents a clinical challenge. Although some clinical and biologic features have been found to be associated with a lower probability of response to imatinib, at present no precise markers allowing for the prediction of outcome for individual patients exist. The most common mechanisms of resistance to imatinib include BCR-ABL kinase domain mutations, amplification, and overexpression of the BCR-ABL oncogene, and clonal evolution with activation of additional transformation pathways. These mechanisms are eventually caused by the genomic instability, which characterizes the Philadelphia chromosome-positive clone. Several approaches to overcome resistance have been proposed. The understanding of at least some of the mechanisms of resistance to imatinib has led to a rapid development of new therapeutic agents that might overcome this resistance. Novel targeted agents designed to overcome imatinib resistance include second-generation TKIs such as dasatinib, nilotinib, bosutinib, bafetinib, and others. Other approaches are exploring combination therapy, with agents affecting different oncogenic pathways, and immune modulation. Herein, we review some of these targeted therapies, particularly those for which clinical data are already available. PMID- 20007108 TI - Molecular monitoring of patients with chronic myeloid leukemia: clinical examples from a non-trial setting. AB - The molecular monitoring of chronic myeloid leukemia allows the clinician a minimally invasive method to judge response to tyrosine kinase therapy and to predict outcome and relapse. Because there are several treatment options for patients with suboptimal response, the ability to proactively predict and respond to relapse makes the "personalization" of treatment a realizable goal. There are practical issues with molecular monitoring, however, including availability of assays, standardization of tests, and the learning curve as doctors and patients learn to follow BCR-ABL levels with interest and reason. This review will examine the use of molecular monitoring in the non-trial setting, concentrating on pitfalls that can occur in the real-world delivery of complex medical care. PMID- 20007109 TI - The next generation of therapies for chronic myeloid leukemia. AB - Therapy with the tyrosine kinase inhibitor (TKI) represents the current standard first-line therapy for the management of patients with chronic myeloid leukemia (CML). Although most patients respond satisfactorily to imatinib, a subset of patients develops resistance mainly because of the acquisition of mutations within the kinase domain of BCR-ABL1 that impair the ability of TKIs to block the activity of the enzyme. Moreover, BCR-ABL1 transcripts can be detected in most patients by molecular techniques, underscoring the limitations of imatinib to eradicate minimal residual disease. Although the resistance conferred by most BCR ABL1 mutations can be overcome with the use of second-generation TKIs such as nilotinib, dasastinib, bosutinib, or bafetinib, the T315I mutation, which represents a common resistance pathway in CML, remains unassailable to TKI therapy. We herein discuss current research efforts in 2 areas of vital importance in CML research, the management of patients with imatinib-resistant mutations, with particular emphasis on those carrying T315I, and the eradication of residual disease. PMID- 20007111 TI - AEOH and the literature of environmental and occupational health. PMID- 20007112 TI - Creating environmental and occupational health: a journal and the field it shaped, 1919-2009. PMID- 20007113 TI - The historical development of academic journals in occupational medicine, 1901 2009. AB - Academic journals in a specialist field provide an interesting historical record of its development and progression over time. This article describes the evolution of some major international journals of occupational medicine, including some historical background on their editorial board. As North America, the United Kingdom, and Northern Europe are known to have the highest contribution to scientific production, it was considered appropriate to investigate the main occupational medicine periodicals in these regions. Given the remarkable improvements in Japanese occupational health following the Second World War, it was also considered worthwhile to investigate the two English language journals of occupational medicine from this country. PMID- 20007114 TI - Historical development of the Archives of Environmental & Occupational Health and its predecessor journals, 1919-2009. AB - The Archives of Environmental & Occupational Health (AEOH) has a long and distinguished history that has so far involved a total of 7 journal titles since 1919. This article provides a detailed historical review of the AEOH, from its seminal precursors of the early 20th century, into the distinguished periodical of today. As editorial leadership is known to influence journal quality, considerable discussion focuses on the careers and professional achievements of the Editors-in-Chief, as well as key members of the editorial board who served the AEOH and its predecessor journals over the past 90 years. PMID- 20007115 TI - Highly cited articles in environmental and occupational health, 1919-1960. AB - Although numerous lists of "citation classics" have been compiled across a variety of scientific fields, few have included articles from environmental and occupational health (EOH). This investigation sought to identify and analyze the most highly cited articles ever published in the Journal of Industrial Hygiene (1919-1935), the Journal of Industrial Hygiene and Toxicology (1936-1949), the Archives of Industrial Hygiene and Occupational Medicine (1950), the American Medical Association (A.M.A.) Archives of Industrial Hygiene and Occupational Medicine (1950-1954), and the A.M.A. Archives of Industrial Health (1955-1960). Regularly cited topics included metal fume fever and various studies of beryllium, whereas the most highly cited article of all was a 1957 paper describing the control of heat casualties at military training centers. Interestingly, the most highly cited articles were not the oldest, and nor were they written as literature reviews. Overall, this study suggests that although some citation patterns in EOH reflect those of other disciplines, the trend is not uniform and EOH itself appears to have some distinctive bibliometric characteristics. PMID- 20007116 TI - A 30-year citation analysis of bibliometric trends at the Archives Of Environmental Health, 1975-2004. AB - This article describes a 30-year citation analysis of the Archives of Environmental Health (AEH), from the earliest available data in 1975, to 2004, when it became the Archives of Environmental & Occupational Health (AEOH). Longitudinal trends were examined regarding the number of items published, the number of citations received, the immediacy index, and the journal's impact factor. A list of the 5 most highly cited articles was also established, including citation frequency and citation lag times. Overall, this study demonstrates that citation analysis can provide an interesting look at the development of a journal over time. The examination of what articles, themes, and topics were being published, cited, or ignored also offers a unique insight into the direction of not only a particular journal, but also the discipline within which it exists. PMID- 20007117 TI - Training in occupational and environmental medicine: the US should look to international models. PMID- 20007118 TI - The direct impact of climate change on regional labor productivity. AB - Global climate change will increase outdoor and indoor heat loads, and may impair health and productivity for millions of working people. This study applies physiological evidence about effects of heat, climate guidelines for safe work environments, climate modeling, and global distributions of working populations to estimate the impact of 2 climate scenarios on future labor productivity. In most regions, climate change will decrease labor productivity, under the simple assumption of no specific adaptation. By the 2080s, the greatest absolute losses of population-based labor work capacity (in the range 11% to 27%) are seen under the A2 scenario in Southeast Asia, Andean and Central America, and the Caribbean. Increased occupational heat exposure due to climate change may significantly impact on labor productivity and costs unless preventive measures are implemented. Workers may need to work longer hours, or more workers may be required, to achieve the same output and there will be economic costs of lost production and/or occupational health interventions against heat exposures. PMID- 20007119 TI - Uranium in drinking water: renal effects of long-term ingestion by an aboriginal community. AB - The authors conducted a study of an aboriginal community to determine if kidney func-tion had been affected by the chronic ingestion of uranium in drinking water from the community's drilled wells. Uranium concentrations in drinking water varied from < 1 to 845 ppb. This nonin-vasive study relied on the measurement of a combination of urinary indicators of kidney function and markers for cell toxicity. In all, 54 individuals (12-73 years old) participated in the study. Correlation of uranium excreted in urine with bio-indicators at p 0.05) after adjustment for co-morbidity. CONCLUSIONS: frailty is not uncommon even among community dwelling young-old men and women in the UK. There are social inequalities in frailty which appear to be mediated by co-morbidity. PMID- 20007128 TI - Hyponatraemia induced by low-dose intravenous pulse cyclophosphamide. AB - BACKGROUND: Cyclophosphamide is an alkylating agent and was traditionally known to potentiate the renal action of vasopressin. Although low-dose intravenous pulse cyclophosphamide therapy is being used extensively in the treatment of malignant and rheumatological diseases, there have been only a few case reports of cyclophosphamide-induced hyponatraemia. METHODS: Clinical data were retrospectively analysed from 84 patients (42 lupus nephritis; 42 non-Hodgkin's lymphoma; a total of 112 treatment episodes) admitted for intravenous pulse cyclophosphamide (500-750 mg/m(2)) therapy. In all patients, half-isotonic saline was used for prophylactic hydration. Cyclophosphamide-induced hyponatraemia was defined as serum sodium concentration <135 mEq/L at 24 hours after the therapy in patients whose basal serum sodium concentrations were normal. RESULTS: After the low-dose intravenous pulse cyclophosphamide, serum sodium concentration significantly decreased from 139.9 +/- 3.5 to 137.9 +/- 5.1 mEq/L (P < 0.001). Cyclophosphamide-induced hyponatraemia occurred in 15 treatment episodes (13.4%) from 12 patients (14.3%). Patients with hyponatraemia were significantly older than those without hyponatraemia (57.3 +/- 14.7 vs. 40.0 +/- 17.0 years, P < 0.01). Hyponatraemia was associated with male sex on univariate analysis (P < 0.05), but not on multivariate analysis. No factors were found to independently predict the occurrence of cyclophosphamide-induced hyponatraemia when multivariate analysis was performed including parameters age, sex, underlying disease, presence or absence of comorbidities associated with hyponatraemia, presence or absence of concurrent medications associated with hyponatraemia and dose of cyclophosphamide. CONCLUSIONS: Hyponatraemia occurring after low-dose intravenous pulse cyclophosphamide is not rare, especially when hypotonic solutions are adopted for hydration protocol. Thus, the use of hypotonic fluids should be avoided when using cyclophosphamide. Instead, isotonic solutions should be used if a forced diuresis is required. PMID- 20007129 TI - Using RIFLE criteria to evaluate acute kidney injury in brain-deceased kidney donors. AB - BACKGROUND: The limited supply of deceased donors for renal transplantation led to considering alternative strategies for making more organs available. One of these strategies is the use of donors with renal dysfunction, as this is usually a reversible condition. RIFLE (risk, injury, failure, loss and end-stage renal failure) criteria were developed to standardize the definition and severity of acute kidney injury (AKI) but have not been previously used in brain-deceased donors. We applied the RIFLE classification to evaluate renal function changes in our donor pool, in an attempt to know its influence in transplant outcome. METHODS: Data were collected from the renal transplant patient and the intensive care unit brain-dead donors prospectively maintained databases of our hospital. Risk was defined when creatinine increased x1.5, injury when it increased x2 and failure when last creatinine increased x3 with respect to admission-day creatinine. RESULTS: From 176 donors, 10.8% suffered AKI and 7.9% were included in 'risk', 2.3% in 'injury' and 0.6% in 'failure' categories. There were no significant differences between AKI and non-AKI groups in donor and intensive care management variables, except in last-day creatinine. First-day urine volumes were lower (P = 0.043) and delayed graft function rates were higher (P = 0.013) in the AKI group than in the non-AKI group recipients. Graft survival and other outcome variables were not different between AKI and non-AKI recipients. CONCLUSIONS: The RIFLE classification system offers us an opportunity to standardize and quantify renal injury in donors. Although >10% of brain-deceased donors can suffer AKI, these grafts can perform adequately. Hence, the development of AKI in donors cannot be an isolated criterion to discard kidney donation. PMID- 20007130 TI - Comparison of alternative methods for scaling dialysis dose. AB - BACKGROUND: Kt/Vurea was established as an index of haemodialysis (HD) adequacy. The use of Vurea as a normalizing factor has been questioned, and alternative parameters such as body weight(0.67) (W(0.67)), body surface area (BSA), resting energy expenditure (REE), high metabolic rate organ (HMRO) mass, liver size (LV) and more recently, bioelectrical resistance (R), an independent and directly measurable biological parameter, were proposed as alternative methods for scaling dialysis dose. METHODS: The present study aimed to prospectively evaluate the predictive power of some demographic, anthropometric, bioelectrical (BIA) and biochemical parameters, of seven scaling parameters, namely Vurea, as derived from the Watson et al. formulae, W(0.67), BSA, REE, HMRO, LV and R and of eight HD adequacy indices [single-pool variable-volume Kt/Vurea, computed using the Daugirdas equation, its rescaled equivalents (Kt/W(0.67), Kt/BSA, Kt/REE, Kt/HMRO, Kt/LV and Kt/R) and Kt] on long-term survival of a cohort of 328 incident white HD patients. All individuals underwent periodical (every 3 months) biochemical evaluations and single-frequency BIA measurements, injecting 800 microA at 50 kHz alternating sinusoidal current with a standard tetrapolar technique. RESULTS: A first Cox regression analysis, testing the predictive power of some demographic, anthropometric, BIA and biochemical parameters, and of the eight HD adequacy indices on long-term survival of the patients, showed that only higher serum creatinine (Scr) levels (P < 0.0001) and lower Kt/R values (P < 0.04) were significant outcome predictors. As Kt was shown not to be an outcome predictor, a second Cox regression analysis, testing the predictive power of the same demographic, anthropometric, BIA and biochemical parameters, and of the seven scaling parameters on long-term survival of the patients, was built. It showed that only higher Scr levels (P < 0.0001) and higher R values (P < 0.04) were significant outcome predictors. Kaplan-Meier survival analyses of the patients stratified into two groups, respectively, according to the first quartile of R values (0.0-467.8 Ohm), the fourth quartile of Kt/R values (98-106 ml/Ohm) and the first quartile of Scr levels (0.0-11.6 mg/dl) showed a significantly higher long-term survival in the groups of patients having R values above the first quartile (P < 0.04), Kt/R values below the fourth quartile (P < 0.03) and Scr levels above the first quartile (P < 0.0001). CONCLUSIONS: Kt/R, R and Scr were independent significant predictors of long-term-survival in incident HD patients: R is related to the fluid status, whereas Scr, which reflects the lean body mass, seems to suggest that body composition is more important than body weight and/or body mass index. Further work is required to develop these concepts and to translate them into rigorous outcome-based adequacy targets suitable for clinical usage. PMID- 20007131 TI - Thermal acclimation of leaf dark respiration of beech seedlings experiencing summer drought in high and low light environments. AB - Little is known about how environmental factors shape the short- and long-term responses of leaf respiration to temperature under field conditions despite the importance of respiration for plant and stand carbon balances. Impacts of water availability and canopy cover on leaf dark respiration (R) and temperature sensitivity were assessed in beech (Fagus sylvatica L.) seedlings in a sub Mediterranean population. We studied seedlings established within canopy gaps (39% global site factor; GSF) that were subject to either no watering (unwatered plants; UW) or regular watering (2-10% higher volumetric topsoil water content as summer progressed; W plants) and seedlings established beneath the adjacent understorey (12% GSF). Leaf R rose exponentially with diurnal increases in temperature; the same temperature sensitivity (Q(10): 2.2) was found for understorey and gap plants, irrespective of watering treatment. Respiration estimated at 25 degrees C (R(25)) was lower in the understorey than the gaps and was significantly lower in the unwatered than in the watered gap plants by the end of summer (0.65 versus 0.80 micromol m(-2) s(-1)). R(25) declined with increasing summer temperature in all plants; however, respiration estimated at the prevailing ambient temperature did not change through the summer. There were parallel declines in R(25) and concentrations of starch and soluble sugars with increasing summer temperature for gap plants. We conclude that seasonal shifts in temperature-response curves of beech leaf R occur in both low- and high-light environments; since leaf R decreased with increasing plant water deficit, such shifts are likely to be greater whenever plants experience summer drought compared to scenarios where plants experience high rainfall in summer. PMID- 20007132 TI - Low soil temperature inhibits the effect of high nutrient supply on photosynthetic response to elevated carbon dioxide concentration in white birch seedlings. AB - To investigate the interactive effects of soil temperature (T(soil)) and nutrient availability on the response of photosynthesis to elevated atmospheric carbon dioxide concentration ([CO(2)]), white birch (Betula papyrifera Marsh.) seedlings were exposed to ambient (360 micromol mol(-1)) or elevated (720 micromol mol(-1)) [CO(2)], three T(soil) (5, 15 and 25 degrees C initially, increased to 7, 17 and 27 degrees C, respectively, 1 month later) and three nutrient regimes (4/1.8/3.3, 80/35/66 and 160/70/132 mg l(-1) N/P/K) for 3 months in environment-controlled greenhouses. Elevated [CO(2)] increased net photosynthetic rate (A(n)), instantaneous water-use efficiency (IWUE), internal to ambient carbon dioxide concentration ratio (C(i)/C(a)), triose phosphate utilization (TPU) and photosynthetic linear electron transport to carboxylation (J(c)), and it decreased actual photochemical efficiency of photosystem II (DeltaF/F(m)'), the fraction of total linear electron transport partitioned to oxygenation (J(o)/J(T)) and leaf N concentration. The low T(soil) suppressed A(n), transpiration rate (E), TPU, DeltaF/F(m)' and J(c), but it increased J(o)/J(T). The low nutrient treatment reduced A(n), IWUE, maximum carboxylation rate of Rubisco, light-saturated electron transport rate, TPU, DeltaF/F(m)', J(c) and leaf N concentration, but increased C(i)/C(a). There were two-factor interactions for C(i)/C(a), TPU and leaf N concentration, and a significant effect of CO(2) x T(soil) x nutrient regime on A(n), IWUE and J(c). The stimulations of A(n) and IWUE by elevated [CO(2)] were limited to seedlings grown under the intermediate and high nutrient regimes at the intermediate and high T(soil). For J(c), the [CO(2)] effect was significant only at intermediate T(soil) + high nutrient availability. No significant [CO(2)] effects were observed under the low T(soil) at any nutrient level. Our results support this study's hypothesis that low T(soil) would reduce the positive effect of high nutrient supply on the response of A(n) to elevated [CO(2)]. PMID- 20007133 TI - Expenditure on obstetric care and the protective effect of insurance on the poor: lessons from two Indonesian districts. AB - OBJECTIVE: The Indonesian Government recently introduced a health insurance scheme to improve access to care for the poor. We investigated the payments made by households for different types of obstetric care, the economic consequences of payments and the effects of the new insurance on that expenditure. METHODS: Expenditures on obstetric care for women were collected from three main hospitals in the Serang and Pandeglang districts of Banten Province for all 'near-miss' cases (372), a sample of normal deliveries (146) and deliveries with Caesarean section (98) over a 6-month period. Women were also interviewed after they were discharged to collect information on economic status, household expenditure and source of payment for care. FINDINGS: Average expenditure by the mothers for near miss cases was found to be Rp 2.6m (US$279) and Rp 1.9m (US$205) in Serang and Pandeglang Hospitals, respectively. Caesarean section was found to be the most expensive intervention. Insurance for the poor covered 51% of women at Serang Hospital and 73% of women at Pandeglang. Around 68% of households in the poorest quintiles would have made catastrophic payments. Insurance for the poor appears to have some positive association with the hospitals' expenditure for treatment of different types of maternal care. CONCLUSION: Insurance for the poor appeared to be relatively effective in protecting households from catastrophic payments. However, it is not sufficient only to cover the very poor; the non-poor can also suffer catastrophic payments and they are only protected because hospital rules over who qualifies have been relaxed. Although the association between insurance and expenditure for obstetric care was important, it was not clear that this represents over-provision of services but rather that it reflected previously inadequate treatment given to those unable to pay. PMID- 20007134 TI - HAMP promoter mutation nc.-153C>T in non p.C282Y homozygous patients with iron overload. PMID- 20007135 TI - Prophylaxis of invasive aspergillosis with voriconazole or caspofungin during building work in patients with acute leukemia. AB - BACKGROUND: Invasive aspergillosis is a common life-threatening infection in patients with acute leukemia. The presence of building work near to hospital wards in which these patients are cared for is an important risk factor for the development of invasive aspergillosis. This study assessed the impact of voriconazole or caspofungin prophylaxis in patients undergoing induction chemotherapy for acute leukemia in a hematology unit exposed to building work. DESIGN AND METHODS: This retrospective cohort study was carried out between June 2003 and January 2006 during which building work exposed patients to a persistently increased risk of invasive aspergillosis. This study compared the cumulative incidence of invasive aspergillosis in patients who did or did not receive primary antifungal prophylaxis. The diagnosis of invasive aspergillosis was based on the European Organization for Research and Treatment of Cancer/Mycosis Study Group criteria. RESULTS: Two-hundred and fifty-seven patients (213 with acute myeloid leukemia, 44 with acute lymphocytic leukemia) were included. The mean age of the patients was 54 years and the mean duration of their neutropenia was 21 days. Eighty-eight received antifungal prophylaxis, most with voriconazole (n=74). The characteristics of the patients who did or did not receive prophylaxis were similar except that pulmonary antecedents (chronic bronchopulmonary disorders or active tobacco use) were more frequent in the prophylaxis group. Invasive aspergillosis was diagnosed in 21 patients (12%) in the non-prophylaxis group and four (4.5%) in the prophylaxis group (P=0.04). Pulmonary antecedents, neutropenia at diagnosis and acute myeloid leukemia with high-risk cytogenetics were positively correlated with invasive aspergillosis, whereas primary prophylaxis was negatively correlated. Survival was similar in both groups. No case of zygomycosis was observed. The 3-month mortality rate was 28% in patients with invasive aspergillosis. CONCLUSIONS: This study suggests that antifungal prophylaxis with voriconazole could be useful in acute leukemia patients undergoing first remission-induction chemotherapy in settings in which there is a high-risk of invasive aspergillosis. PMID- 20007136 TI - Homozygous deletion of HFE is the common cause of hemochromatosis in Sardinia. PMID- 20007137 TI - Tyrosine kinase inhibitor therapy can cure chronic myeloid leukemia without hitting leukemic stem cells. AB - BACKGROUND: Tyrosine kinase inhibitors, such as imatinib, are not considered curative for chronic myeloid leukemia--regardless of the significant reduction of disease burden during treatment--since they do not affect the leukemic stem cells. However, the stochastic nature of hematopoiesis and recent clinical observations suggest that this view must be revisited. DESIGN AND METHODS: We studied the natural history of a large cohort of virtual patients with chronic myeloid leukemia under tyrosine kinase inhibitor therapy using a computational model of hematopoiesis and chronic myeloid leukemia that takes into account stochastic dynamics within the hematopoietic stem and early progenitor cell pool. RESULTS: We found that in the overwhelming majority of patients the leukemic stem cell population undergoes extinction before disease diagnosis. Hence leukemic progenitors, susceptible to tyrosine kinase inhibitor attack, are the natural target for chronic myeloid leukemia treatment. Response dynamics predicted by the model closely match data from clinical trials. We further predicted that early diagnosis together with administration of tyrosine kinase inhibitor opens the path to eradication of chronic myeloid leukemia, leading to the wash out of the aberrant progenitor cells, ameliorating the patient's condition while lowering the risk of blast transformation and drug resistance. CONCLUSIONS: Tyrosine kinase inhibitor therapy can cure chronic myeloid leukemia, although it may have to be prolonged. The depth of response increases with time in the vast majority of patients. These results illustrate the importance of stochastic effects on the dynamics of acquired hematopoietic stem cell disorders and have direct relevance for other hematopoietic stem cell-derived diseases. PMID- 20007138 TI - Complications and treatment of patients with beta-thalassemia in France: results of the National Registry. AB - BACKGROUND: beta-thalassemia is a rare disease in France, encountered mainly in patients originating from Italy and North Africa. In the setting of the recent French plan for rare diseases, a National Registry for thalassemia has been developed since 2005. Epidemiological and clinical data have been collected on living patients with beta-thalassemia major or intermedia, including those who underwent hematopoietic stem cell transplantation. DESIGN AND METHODS: A standardized questionnaire was sent to clinicians throughout the national professional networks involved in the management of thalassemic patients and data were updated every 18 months. A cross-sectional study was performed in February 2009. RESULTS: Data on 378 patients (267 with thalassemia major) with a median age of 20 were recorded. Hematopoietic stem cell transplantation was performed in 52 patients. Stature, rates of parenthood, splenectomy, and cholecystectomy were no different between non-transplanted thalassemia major and thalassemia intermedia patients, after adjustment for age. Among the 215 non-transplanted thalassemia major patients, the median serum ferritin level was 1240 ng/mL and the rates of iron-related complications were 10%, 6%, 10% and 48% for cardiac failure, diabetes, hypothyroidism, and hypogonadism, respectively. From 2005 to 2008, a dramatic switch in chelation treatment, from deferoxamine to deferasirox, was observed. CONCLUSIONS: The rates of complications of iron overload in French thalassemia major patients appeared similar to those reported in other developed countries in which this condition is not endemic. There were no significant differences in height and parenthood rates between patients with the major and the intermedia forms of the disease, underlining the progress in clinical care. Future developments will focus on mortality and morbidity under oral chelation treatment. PMID- 20007139 TI - Autocrine IGF-1/IGF-1R signaling is responsible for constitutive PI3K/Akt activation in acute myeloid leukemia: therapeutic value of neutralizing anti-IGF 1R antibody. AB - BACKGROUND: Alterations in the PI3K/Akt pathway are found in a wide range of cancers and the development of PI3K inhibitors represents a promising approach to cancer therapy. Constitutive PI3K activation, reflecting an intrinsic oncogenic deregulation of primary blast cells, is detected in 50% of patients with acute myeloid leukemia. However, the mechanisms leading to this activation are currently unknown. As we previously reported IGF-1 autocriny in acute myeloid leukemia cells, we investigated whether IGF-1 signaling was involved in the constitutive activation of PI3K. DESIGN AND METHODS: We analyzed the IGF-1/IGF-1R signaling pathway and PI3K activity in 40 acute myeloid leukemia bone marrow samples. Specific inhibition of IGF-1/IGF-1R signaling was investigated using neutralizing anti-IGF-1R, anti-IGF-1 antibodies or IGF-1 short interfering RNA. The anti-leukemic activity of the neutralizing anti-IGF-1R was tested by analyzing its effects on leukemic progenitor clonogenicity, blast cell proliferation and survival. RESULTS: In all samples tested, we found that functional IGF-1R was constantly expressed in leukemic cells. In the acute myeloid leukemia samples with PI3K activation, we found that the IGF-1R was constitutively phosphorylated, although no IGF-1R activating mutation was detected. Specific inhibition of IGF-1R signaling with neutralizing anti-IGF-1R strongly inhibited the constitutive phosphorylation of both IGF-1R and Akt in 70% of the PI3K activated samples. Moreover, both incubation with anti-IGF-1 antibody and IGF-1 short interfering RNA inhibited Akt phosphorylation in leukemic cells. Finally, neutralizing anti-IGF-1R treatment decreased the clonogenicity of leukemic progenitors and the proliferation of PI3K activated acute myeloid leukemia cells. CONCLUSIONS: Our current data indicate a critical role for IGF-1 autocriny in constitutive PI3K/Akt activation in primary acute myeloid leukemia cells and provide a strong rationale for targeting IGF-1R as a potential new therapy for this disease. PMID- 20007140 TI - Interleukin-3 promotes hemangioblast development in mouse aorta-gonad-mesonephros region. AB - BACKGROUND: The hemangioblast is a bi-potential precursor cell with the capacity to differentiate into hematopoietic and vascular cells. In mouse E7.0-7.5 embryos, the hemangioblast can be identified by a clonal blast colony-forming cell (BL-CFC) assay or single cell OP9 co-culture. However, the ontogeny of the hemangioblast in mid-gestation embryos is poorly defined. DESIGN AND METHODS: The BL-CFC assay and the OP9 system were combined to illustrate the hemangioblast with lymphomyeloid and vascular potential in the mouse aorta-gonad-mesonephros region. The colony-forming assay, reverse transcriptase polymerase chain reaction analysis, immunostaining and flow cytometry were used to identify the hematopoietic potential, and Matrigel- or OP9-based methods were employed to evaluate endothelial progenitor activity. RESULTS: Functionally, the aorta-gonad mesonephros-derived BL-CFC produced erythroid/myeloid progenitors, CD19(+) B lymphocytes, and CD3(+)TCRbeta(+) T lymphocytes. Meanwhile, the BL-CFC-derived adherent cells generated CD31(+) tube-like structures on OP9 stromal cells, validating the endothelial progenitor potential. The aorta-gonad-mesonephros derived hemangioblast was greatly enriched in CD31(+), endomucin(+) and CD105(+) subpopulations, which collectively pinpoints the endothelial layer as the main location. Interestingly, the BL-CFC was not detected in yolk sac, placenta, fetal liver or embryonic circulation. Screening of candidate cytokines revealed that interleukin-3 was remarkable in expanding the BL-CFC in a dose-dependent manner through the JAK2/STAT5 and MAPK/ERK pathways. Neutralizing interleukin-3 in the aorta-gonad-mesonephros region resulted in reduced numbers of BL-CFC, indicating the physiological requirement for this cytokine. Both hematopoietic and endothelial differentiation potential were significantly increased in interleukin 3-treated BL-CFC, suggesting a persistent positive influence. Intriguingly, interleukin-3 markedly amplified primitive erythroid and macrophage precursors in E7.5 embryos. Quantitative polymerase chain reaction analysis demonstrated declined Flk-1 and elevated Scl and von Willebrand factor transcription upon interleukin-3 stimulation, indicating accelerated hemangiopoiesis. CONCLUSIONS: The hemangioblast with lymphomyeloid potential is one of the precursors of definitive hematopoiesis in the mouse aorta-gonad-mesonephros region. Interleukin 3 has a regulatory role with regards to both the number and capacity of the dual potential hemangioblast. PMID- 20007141 TI - Erythrocytosis associated with a novel missense mutation in the HIF2A gene. AB - The ERYTHROPOIETIN (EPO) gene is regulated by the transcription factor Hypoxia Inducible Factor-alpha (HIF-alpha). In this pathway, Prolyl Hydroxylase Domain protein 2 (PHD2) hydroxylates two prolyl residues in HIF-alpha, which in turn promotes HIF-alpha degradation by the von Hippel Lindau (VHL) protein. Evidence that HIF-2alpha is the important isoform for EPO regulation in humans comes from the recent observation that mutations in the HIF2A gene are associated with cases of erythrocytosis. We report here a new erythrocytosis-associated mutation, p.Asp539Glu, in the HIF2A gene. Similar to all reported cases, the affected residue is in close vicinity and C-terminal to the primary hydroxylation site in HIF-2alpha, Pro531. This mutation, however, is notable in producing a rather subtle amino acid substitution. Nonetheless, we find that this mutation compromises binding of HIF-2alpha to both PHD2 and VHL, and we propose that this mutation is the cause of erythrocytosis in this individual. PMID- 20007142 TI - Different risk of deep vein thrombosis and pulmonary embolism in carriers with factor V Leiden compared with non-carriers, but not in other thrombophilic defects. Results from a large retrospective family cohort study. AB - The term factor V Leiden (FVL) paradox is used to describe the different risk of deep vein thrombosis and pulmonary embolism that has been found in carriers of FVL. In a thrombophilic family-cohort, we estimated differences in absolute risks of deep vein thrombosis and pulmonary embolism for various thrombophilic defects. Of 2,054 relatives, 1,131 were female, 41 had pulmonary embolism and 126 deep vein thrombosis. Annual incidence for deep vein thrombosis in non-carriers of FVL was 0.19% (95%CI, 0.16-0.23), and 0.41% (95%CI, 0.28-0.58) in carriers; relative risk (RR) 2.1 (95%CI, 1.4-3.2). For pulmonary embolism these incidences were similar in carriers and non-carriers 0.07%, respectively; RR 1.0 (95% CI, 0.4 2.5). When co-inheritance of other thrombophilic defects was excluded the RR for deep vein thrombosis in FVL carriers was 7.0 (95%CI, 2.3-21.7) compared to non carriers and 2.8 (95%CI, 0.5-14.4) for pulmonary embolism. For other thrombophilic defects no such effect was observed. Thus the FVL paradox was confirmed in our study. However, a similar paradox in carriers of other thrombophilic defects was not observed. PMID- 20007143 TI - A modified EBMT risk score and the hematopoietic cell transplantation-specific comorbidity index for pre-transplant risk assessment in adult acute lymphoblastic leukemia. AB - BACKGROUND: Disease stage is the most important prognostic parameter in allogeneic hematopoietic cell transplantation (HCT) for acute lymphoblastic leukemia, but other factors such as donor/host histocompatibility and gender combination, recipient age, performance status and comorbidities need to be considered. Several scoring systems are available to predict outcome in HCT recipients; however, their prognostic relevance in acute lymphoblastic leukemia is not well defined. DESIGN AND METHODS: In the present study we evaluated a modified EBMT risk score (mEBMT) and the HCT-specific comorbidity index (HCT-CI) in 151 adult acute lymphoblastic leukemia patients who received allogeneic HCT from 1995 until 2007 at our center. RESULTS: Disease status was first complete remission (CR1) (47%), CR>1 (21%) or no CR (32%). Overall survival (OS) at one, two and five years was 62%, 51% and 40% and non-relapse mortality (NRM) was 21%, 24% and 32%. Median mEBMT was 3 (0-6). Higher mEBMT was associated with inferior OS (hazard ratio per score unit (HR): 1.50, P<0.001), higher NRM (HR: 1.36, P=0.042) and higher relapse mortality (HR: 1.68, P<0.001). Disease stage was the predominant prognostic factor in this score. Comorbidities were present in 71% of patients with mild hepatic disease (29%), moderate pulmonary disease (28%) and infections (23%) being the most common. Median HCT-CI was 1 (0-9). In univariate analysis a trend for inferior OS (HR: 1.08, P=0.20) and higher NRM (HR: 1.14, P=0.11) with increasing HCT-CI was observed but the level of significance was not reached. In additional analyses we found that reduced Karnofsky Performance Status (KPS) was associated with inferior OS (HR: 1.34, P=0.023) and higher relapse mortality (HR: 1.71, P=0.001) when analyzed univariately. However, KPS was associated with disease stage and significance was lost in multivariate analysis. CONCLUSIONS: The mEBMT was prognostic in our patient cohort with predominant influence of disease stage, whereas a trend but no significant prognostic value was observed for the HCT-CI. PMID- 20007145 TI - The -582A>G variant of the HAMP promoter is not associated with high serum ferritin levels in normal subjects. PMID- 20007144 TI - Hematopoietic stem cell transplantation for paroxysmal nocturnal hemoglobinuria: long-term results of a retrospective study on behalf of the Gruppo Italiano Trapianto Midollo Osseo (GITMO). AB - BACKGROUND: Paroxysmal nocturnal hemoglobinuria is an acquired clonal disorder of the hemopoietic stem cells for which the only curative treatment is allogeneic hematopoietic stem cell transplantation. DESIGN AND METHODS: The aim of this retrospective study was to assess the long-term clinical and hematologic results in 26 paroxysmal nocturnal hemoglobinuria patients who received hematopoietic stem cell transplantation in Italy between 1988 and 2006. The patients were aged 22 to 60 years (median 32 years). Twenty-three donors were HLA-identical (22 siblings and one unrelated) and 3 were HLA-mismatched (2 related and one unrelated). RESULTS: Fifteen patients received a myeloablative conditioning consisting of busulfan and cyclophosphamide (in all cases from identical donor) and 11 were given a reduced intensity conditioning (8 from identical donor and 3 from mismatched donor). The cumulative incidence of graft failure was 8% (4% primary and 4% secondary graft failure). Transplant-related mortality for all patients was 42% (26% and 63% for patients transplanted following myeloablative or reduced intensity conditioning, respectively). As of October 31, 2009, 15 patients (11 in the myeloablative conditioning group and 4 in the reduced intensity conditioning group) are alive with complete hematologic recovery and no evidence of paroxysmal nocturnal hemoglobinuria following a median follow-up of 131 months (range 30-240). The 10-year Kaplan-Meier probability of disease-free survival was 57% for all patients: 65% for 23 patients transplanted from identical donor and 73% for 15 patients transplanted with myeloablative conditioning. No thromboembolic event nor recurrence of the disease were reported following transplant. CONCLUSIONS: The findings of this study confirm that most patients with paroxysmal nocturnal hemoglobinuria may be definitively cured with hematopoietic stem cell transplantation. PMID- 20007146 TI - Chimeric tRNAs as tools to induce proteome damage and identify components of stress responses. AB - Misfolded proteins are caused by genomic mutations, aberrant splicing events, translation errors or environmental factors. The accumulation of misfolded proteins is a phenomenon connected to several human disorders, and is managed by stress responses specific to the cellular compartments being affected. In wild type cells these mechanisms of stress response can be experimentally induced by expressing recombinant misfolded proteins or by incubating cells with large concentrations of amino acid analogues. Here, we report a novel approach for the induction of stress responses to protein aggregation. Our method is based on engineered transfer RNAs that can be expressed in cells or tissues, where they actively integrate in the translation machinery causing general proteome substitutions. This strategy allows for the introduction of mutations of increasing severity randomly in the proteome, without exposing cells to unnatural compounds. Here, we show that this approach can be used for the differential activation of the stress response in the Endoplasmic Reticulum (ER). As an example of the applications of this method, we have applied it to the identification of human microRNAs activated or repressed during unfolded protein stress. PMID- 20007148 TI - ViralORFeome: an integrated database to generate a versatile collection of viral ORFs. AB - Large collections of protein-encoding open reading frames (ORFs) established in a versatile recombination-based cloning system have been instrumental to study protein functions in high-throughput assays. Such 'ORFeome' resources have been developed for several organisms but in virology, plasmid collections covering a significant fraction of the virosphere are still needed. In this perspective, we present ViralORFeome 1.0 (http://www.viralorfeome.com), an open-access database and management system that provides an integrated set of bioinformatic tools to clone viral ORFs in the Gateway(R) system. ViralORFeome provides a convenient interface to navigate through virus genome sequences, to design ORF-specific cloning primers, to validate the sequence of generated constructs and to browse established collections of virus ORFs. Most importantly, ViralORFeome has been designed to manage all possible variants or mutants of a given ORF so that the cloning procedure can be applied to any emerging virus strain. A subset of plasmid constructs generated with ViralORFeome platform has been tested with success for heterologous protein expression in different expression systems at proteome scale. ViralORFeome should provide our community with a framework to establish a large collection of virus ORF clones, an instrumental resource to determine functions, activities and binding partners of viral proteins. PMID- 20007147 TI - Reduced nuclear export of HuR mRNA by HuR is linked to the loss of HuR in replicative senescence. AB - The RNA-binding protein, HuR, associates with the HuR mRNA, but the consequences of this interaction are unknown. Here, we use human diploid fibroblasts (HDFs) and cervical carcinoma cells to study this regulatory paradigm. Ectopic overexpression of HuR potently enhanced the translation and cytoplasmic levels of endogenous HuR, but did not affect HuR mRNA levels. Inhibition of CRM1 function by Lemptomycin B or by knockdown of CRM1 greatly diminished the cytoplasmic levels of endogenous HuR mRNA and hence blocked the induction of endogenous HuR by exogenous HuR. Further studies showed that HuR interacted with the 3' untranslated region (UTR) of HuR and that overexpression of HuR increased the cytoplasmic levels of a chimeric luciferase-HuR 3'-UTR reporter transcript, as well as luciferase activity; conversely, HuR knockdown reduced both parameters. Moreover, the loss of HuR in senescent, late-passage HDFs was accompanied by a reduced cytoplasmic presence of endogenous HuR mRNA, ectopic Luc-HuR-3'UTR reporter transcript, and luciferase activity relative to what was observed in young, early-passage cells. Our results reveal a positive feedback mechanism for the regulation of HuR, which may play an important role in the regulation of HuR during replicative senescence. PMID- 20007149 TI - Toxin-antitoxin based transgene expression in mammalian cells. AB - Long-term, recombinant gene expression in mammalian cells depends on the nature of the transgene integration site and its inherent properties to modulate transcription (epigenetic effects). Here we describe a method by which high transgene expression is achieved and stabilized in extensively proliferating cultures. The method is based on strict co-expression of the transgene with an antitoxin in cells that express the respective toxin. Since the strength of antitoxin expression correlates with an advantage for cell growth, the cells with strong antitoxin expression are enriched over time in cultures of heterogeneous cells. This principle was applied to CHO cell lines that conditionally express the toxin kid and that are transduced to co-express the antitoxin kis together with different transgenes of interest. Cultivation of pools of transfectants that express the toxin steadily increase their transgene expression within several weeks to reach a maximum that is up to 120-fold over the initial status. In contrast, average transgene expression drops in the absence of toxin expression. Together, we show that cells conditionally expressing kid can be employed to create overexpressing cells by a simple coupling of kis to the transgene of interest, without further manipulation and in absence of selectable drugs. PMID- 20007151 TI - The Protein Naming Utility: a rules database for protein nomenclature. AB - Generation of syntactically correct and unambiguous names for proteins is a challenging, yet vital task for functional annotation processes. Proteins are often named based on homology to known proteins, many of which have problematic names. To address the need to generate high-quality protein names, and capture our significant experience correcting protein names manually, we have developed the Protein Naming Utility (PNU, http://www.jcvi.org/pn-utility). The PNU is a web-based database for storing and applying naming rules to identify and correct syntactically incorrect protein names, or to replace synonyms with their preferred name. The PNU allows users to generate and manage collections of naming rules, optionally building upon the growing body of rules generated at the J. Craig Venter Institute (JCVI). Since communities often enforce disparate conventions for naming proteins, the PNU supports grouping rules into user managed collections. Users can check their protein names against a selected PNU rule collection, generating both statistics and corrected names. The PNU can also be used to correct GenBank table files prior to submission to GenBank. Currently, the database features 3080 manual rules that have been entered by JCVI Bioinformatics Analysts as well as 7458 automatically imported names. PMID- 20007152 TI - An intermolecular RNA triplex provides insight into structural determinants for the pseudoknot stimulator of -1 ribosomal frameshifting. AB - An efficient -1 programmed ribosomal frameshifting (PRF) signal requires an RNA slippery sequence and a downstream RNA stimulator, and the hairpin-type pseudoknot is the most common stimulator. However, a pseudoknot is not sufficient to promote -1 PRF. hTPK-DU177, a pseudoknot derived from human telomerase RNA, shares structural similarities with several -1 PRF pseudoknots and is used to dissect the roles of distinct structural features in the stimulator of -1 PRF. Structure-based mutagenesis on hTPK-DU177 reveals that the -1 PRF efficiency of this stimulator can be modulated by sequential removal of base-triple interactions surrounding the helical junction. Further analysis of the junction flanking base triples indicates that specific stem-loop interactions and their relative positions to the helical junction play crucial roles for the -1 PRF activity of this pseudoknot. Intriguingly, a bimolecular pseudoknot approach based on hTPK-DU177 reveals that continuing triplex structure spanning the helical junction, lacking one of the loop-closure features embedded in pseudoknot topology, can stimulate -1 PRF. Therefore, the triplex structure is an essential determinant for the DU177 pseudoknot to stimulate -1 PRF. Furthermore, it suggests that -1 PRF, induced by an in-trans RNA via specific base-triple interactions with messenger RNAs, can be a plausible regulatory function for non coding RNAs. PMID- 20007150 TI - 5-methylcytosine in RNA: detection, enzymatic formation and biological functions. AB - The nucleobase modification 5-methylcytosine (m(5)C) is widespread both in DNA and different cellular RNAs. The functions and enzymatic mechanisms of DNA m(5)C methylation were extensively studied during the last decades. However, the location, the mechanism of formation and the cellular function(s) of the same modified nucleobase in RNA still remain to be elucidated. The recent development of a bisulfite sequencing approach for efficient m(5)C localization in various RNA molecules puts ribo-m(5)C in a highly privileged position as one of the few RNA modifications whose detection is amenable to PCR-based amplification and sequencing methods. Additional progress in the field also includes the characterization of several specific RNA methyltransferase enzymes in various organisms, and the discovery of a new and unexpected link between DNA and RNA m(5)C-methylation. Numerous putative RNA:m(5)C-MTases have now been identified and are awaiting characterization, including the identification of their RNA substrates and their related cellular functions. In order to bring these recent exciting developments into perspective, this review provides an ordered overview of the detection methods for RNA methylation, of the biochemistry, enzymology and molecular biology of the corresponding modification enzymes, and discusses perspectives for the emerging biological functions of these enzymes. PMID- 20007153 TI - Dependence of aptamer activity on opposed terminal extensions: improvement of light-regulation efficiency. AB - Aptamers that can be regulated with light allow precise control of protein activity in space and time and hence of biological function in general. In a previous study, we showed that the activity of the thrombin-binding aptamer HD1 can be turned off by irradiation using a light activatable 'caged' intramolecular antisense-domain. However, the activity of the presented aptamer in its ON state was only mediocre. Here we studied the nature of this loss in activity in detail and found that switching from 5'- to 3'-extensions affords aptamers that are even more potent than the unmodified HD1. In particular we arrived at derivatives that are now more active than the aptamer NU172 that is currently in phase 2 clinical trials as an anticoagulant. As a result, we present light-regulatable aptamers with a superior activity in their ON state and an almost digital ON/OFF behavior upon irradiation. PMID- 20007154 TI - In vitro selection of oligonucleotides that bind double-stranded DNA in the presence of triplex-stabilizing agents. AB - A SELEX approach has been developed in order to select oligonucleotides that bind double-stranded DNA in the presence of a triplex-stabilizing agent, and was applied to a target sequence containing an oligopurine-oligopyrimidine stretch. After only seven rounds of selection, the process led to the identification of oligonucleotides that were able to form triple helices within the antiparallel motif. Inspection of the selected sequences revealed that, contrary to GC base pair which were always recognized by guanines, recognition of AT base pair could be achieved by either adenine or thymine, depending on the sequence context. While thymines are strongly preferred for several positions, some others can accommodate the presence of adenines. These results contribute to set the rules for designing oligonucleotides that form stable triple helices in the presence of triplex-stabilizing agents at physiological pH. They set the basis for further experiments regarding extension of potential target sequences for triple-helix formation or recognition of ligand-DNA complexes. PMID- 20007158 TI - The interface between metabolic and stress signalling. AB - BACKGROUND: It is becoming increasingly clear that stress and metabolic signalling networks interact and that this interaction is important in plant responses to herbivory, pathogen attack, drought, cold, heat and osmotic stresses including salinity. At the interface between these two major signalling systems are the hormone abscisic acid (ABA) and signalling factors including protein kinases and transcription factors. SCOPE: This briefing reviews links between ABA, stress and sugar signalling, focusing on the roles of sucrose non-fermenting 1-related protein kinases (SnRKs), SnRK1-activating protein kinases (SnAKs), calcium-dependent protein kinases (CDPKs) and ABA response element binding proteins (AREBPs, which are transcription factors). Links between stress and nitrogen / amino acid signalling are also described, including the roles of a protein kinase called general control non-derepressible (GCN)-2 in regulating protein synthesis through phosphorylation of the alpha-subunit of translation initiation factor-2 (eIF2alpha) in response not only to decreases in amino acid levels but also to a range of stresses. Evidence of a link between sugar and amino acid signalling is explored, with nitrate reductase being a target for regulation by both SnRK1 and GCN2 through different mechanisms; possible links between SnRK1 and GCN2 via a pathway including the protein kinase target of rapamycin (TOR)-1 are described. The significance of these interactions to the concept of signalling networks as opposed to simple cascades and pathways, and the importance of the subject in the context of the predicted increase in severity and range of stresses that plants will have to withstand as a result of global climate change are discussed. PMID- 20007159 TI - Has the frequency of bleeding changed over time for patients presenting with an acute coronary syndrome? The global registry of acute coronary events. AB - AIMS: To determine whether changes in practice, over time, are associated with altered rates of major bleeding in acute coronary syndromes (ACS). METHODS AND RESULTS: Patients from the Global Registry of Acute Coronary Events were enrolled between 2000 and 2007. The main outcome measures were frequency of major bleeding, including haemorrhagic stroke, over time, after adjustment for patient characteristics, and impact of major bleeding on death and myocardial infarction. Of the 50 947 patients, 2.3% sustained a major bleed; almost half of these presented with ST-elevation ACS (44%, 513). Despite changes in antithrombotic therapy (increasing use of low molecular weight heparin, P < 0.0001), thienopyridines (P < 0.0001), and percutaneous coronary interventions (P < 0.0001), frequency of major bleeding for all ACS patients decreased (2.6 to 1.8%; P < 0.0001). Most decline was seen in ST-elevation ACS (2.9 to 2.1%, P = 0.02). The overall decline remained after adjustment for patient characteristics and treatments (P = 0.002, hazard ratio 0.94 per year, 95% confidence interval 0.91 0.98). Hospital characteristics were an independent predictor of bleeding (P < 0.0001). Patients who experienced major bleeding were at increased risk of death within 30 days from admission, even after adjustment for baseline variables. CONCLUSION: Despite increasing use of more intensive therapies, there was a decline in the rate of major bleeding associated with changes in clinical practice. However, individual hospital characteristics remain an important determinant of the frequency of major bleeding. PMID- 20007160 TI - Low prevalence of atrial fibrillation in asymptomatic adults in Geneva, Switzerland. AB - AIMS: To determine the prevalence of atrial fibrillation (AF) in a population based sample of adults. METHODS AND RESULTS: Between January 2005 and December 2007 individuals aged > or =50 years, residents of the city of Geneva, who had participated in a previous random survey were invited for follow-up examination. AF was assessed on a single resting 6-lead ECG. Reported prevalences were standardized for the age distribution of Canton Geneva. Overall participation was 72.8%. Twenty-nine cases of AF (22 men) were diagnosed among 3285 subjects (1696 men). The crude prevalence of AF (95% CI) was 0.88% (0.86, 0.90) overall, but higher in men [1.30% (1.26, 1.34)] than in women [0.44% (0.41, 0.47)]. The age standardized AF prevalence was slightly higher [overall: 0.94% (0.91, 0.97), men: 1.23% (1.19, 1.27), women: 0.54% (0.47, 0.61)]. AF prevalence increased with age in both sexes. A 'history of suspected arterial embolism' (brain or legs) was higher in the AF cases (10.3 vs. 3.3%; P = 0.03). CONCLUSION: This population based survey of a general Swiss population indicates that the prevalence of AF remains below 1%. These results are less alarming than those from previous studies based on patients seeking medical care. PMID- 20007161 TI - Non-invasive diagnosis of endometriosis based on a combined analysis of six plasma biomarkers. AB - BACKGROUND: Lack of a non-invasive diagnostic test contributes to the long delay between onset of symptoms and diagnosis of endometriosis. The aim of this study was to evaluate the combined performance of six potential plasma biomarkers in the diagnosis of endometriosis. METHODS: This case-control study was conducted in 294 infertile women, consisting of 93 women with a normal pelvis and 201 women with endometriosis. We measured plasma concentrations of interleukin (IL)-6, IL 8, tumour necrosis factor-alpha, high-sensitivity C-reactive protein (hsCRP), and cancer antigens CA-125 and CA-19-9. Analyses were done using the Kruskal-Wallis test, Mann-Whitney test, receiver operator characteristic, stepwise logistic regression and least squares support vector machines (LSSVM). RESULTS: Plasma levels of IL-6, IL-8 and CA-125 were increased in all women with endometriosis and in those with minimal-mild endometriosis, compared with controls. In women with moderate-severe endometriosis, plasma levels of IL-6, IL-8 and CA-125, but also of hsCRP, were significantly higher than in controls. Using stepwise logistic regression, moderate-severe endometriosis was diagnosed with a sensitivity of 100% (specificity 84%) and minimal-mild endometriosis was detected with a sensitivity of 87% (specificity 71%) during the secretory phase. Using LSSVM analysis, minimal-mild endometriosis was diagnosed with a sensitivity of 94% (specificity 61%) during the secretory phase and with a sensitivity of 92% (specificity 63%) during the menstrual phase. CONCLUSIONS: Advanced statistical analysis of a panel of six selected plasma biomarkers on samples obtained during the secretory phase or during menstruation allows the diagnosis of both minimal mild and moderate-severe endometriosis with high sensitivity and clinically acceptable specificity. PMID- 20007162 TI - Improved viability of populations with diverse life-history portfolios. AB - A principle shared by both economists and ecologists is that a diversified portfolio spreads risk, but this idea has little empirical support in the field of population biology. We found that population growth rates (recruits per spawner) and life-history diversity as measured by variation in freshwater and ocean residency were negatively correlated across short time periods (one to two generations), but positively correlated at longer time periods, in nine Bristol Bay sockeye salmon populations. Further, the relationship between variation in growth rate and life-history diversity was consistently negative. These findings strongly suggest that life-history diversity can both increase production and buffer population fluctuations, particularly over long time periods. Our findings provide new insights into the importance of biocomplexity beyond spatio-temporal aspects of populations, and suggest that maintaining diverse life-history portfolios of populations may be crucial for their resilience to unfavourable conditions like habitat loss and climate change. PMID- 20007163 TI - Toxicity of non-steroidal anti-inflammatory drugs to Gyps vultures: a new threat from ketoprofen. AB - Three Gyps vulture species are on the brink of extinction in South Asia owing to the veterinary non-steroidal anti-inflammatory drug (NSAID) diclofenac. Carcasses of domesticated ungulates are the main food source for Asia's vultures and birds die from kidney failure after consuming diclofenac-contaminated tissues. Here, we report on the safety testing of the NSAID ketoprofen, which was not reported to cause mortality in clinical treatment of scavenging birds and is rapidly eliminated from livestock tissues. Safety testing was undertaken using captive non-releasable Cape griffon vultures (Gyps coprotheres) and wild-caught African white-backed vultures (G. africanus), both previously identified as susceptible to diclofenac and suitable surrogates. Ketoprofen doses ranged from 0.5 to 5 mg kg(-1) vulture body weight, based upon recommended veterinary guidelines and maximum levels of exposure for wild vultures (estimated as 1.54 mg kg(-1)). Doses were administered by oral gavage or through feeding tissues from cattle dosed with ketoprofen at 6 mg kg(-1) cattle body weight, before slaughter. Mortalities occurred at dose levels of 1.5 and 5 mg kg(-1) vulture body weight (within the range recommended for clinical treatment) with the same clinical signs as observed for diclofenac. Surveys of livestock carcasses in India indicate that toxic levels of residual ketoprofen are already present in vulture food supplies. Consequently, we strongly recommend that ketoprofen is not used for veterinary treatment of livestock in Asia and in other regions of the world where vultures access livestock carcasses. The only alternative to diclofenac that should be promoted as safe for vultures is the NSAID meloxicam. PMID- 20007164 TI - Temporal learning of predation risk by embryonic amphibians. AB - For prey species that rely on learning to recognize their predators, natural selection should favour individuals able to learn as early as possible. The earliest point at which individuals can gather information about the identity of their potential predators is during the embryonic stage. Indeed, recent experiments have demonstrated that amphibians can learn to recognize predators prior to hatching. Here, we conditioned woodfrog embryos to recognize predatory salamander cues either in the morning or in the evening, and subsequently exposed the two-week-old tadpoles to salamander cues either in the morning or in the evening, and recorded the intensity of their antipredator behaviour. The data indicate that amphibians learn to recognize potential predators while still in the egg, and also learn the temporal component of this information, which they use later in life, to adjust the intensity of their antipredator responses throughout the day. PMID- 20007165 TI - Impending conservation crisis for Southeast Asian amphibians. AB - With an understudied amphibian fauna, the highest deforestation rate on the planet and high harvesting pressures, Southeast Asian amphibians are facing a conservation crisis. Owing to the overriding threat of habitat loss, the most critical conservation action required is the identification and strict protection of habitat assessed as having high amphibian species diversity and/or representing distinctive regional amphibian faunas. Long-term population monitoring, enhanced survey efforts, collection of basic biological and ecological information, continued taxonomic research and evaluation of the impact of commercial trade for food, medicine and pets are also needed. Strong involvement of regional stakeholders, students and professionals is essential to accomplish these actions. PMID- 20007167 TI - Shrewd alliances: mixed foraging associations between treeshrews, greater racket tailed drongos and sparrowhawks on Great Nicobar Island, India. AB - Mixed-species foraging associations may form to enhance feeding success or to avoid predators. We report the costs and consequences of an unusual foraging association between an endemic foliage gleaning tupaid (Nicobar treeshrew Tupaia nicobarica) and two species of birds; one an insectivorous commensal (greater racket-tailed drongo Dicrurus paradiseus) and the other a diurnal raptor and potential predator (Accipiter sp.). In an alliance driven, and perhaps engineered, by drongos, these species formed cohesive groups with predictable relationships. Treeshrew breeding pairs were found more frequently than solitary individuals with sparrowhawks and were more likely to tolerate sparrowhawks in the presence of drongos. Treeshrews maintained greater distances from sparrowhawks than drongos, and permitted the raptors to come closer when drongos were present. Treeshrew foraging rates declined in the presence of drongos; however, the latter may provide them predator avoidance benefits. The choice of the raptor to join the association is intriguing; particular environmental resource states may drive the evolution of such behavioural strategies. Although foraging benefits seem to be the primary driver of this association, predator avoidance also influences interactions, suggesting that strategies driving the formation of flocks may be complex and context dependent with varying benefits for different actors. PMID- 20007166 TI - Plausibility of inferred ancestral phenotypes and the evaluation of alternative models of limb evolution in scincid lizards. AB - Phylogenetic approaches to inferring ancestral character states are becoming increasingly sophisticated; however, the potential remains for available methods to yield strongly supported but inaccurate ancestral state estimates. The consistency of ancestral states inferred for two or more characters affords a useful criterion for evaluating ancestral trait reconstructions. Ancestral state estimates for multiple characters that entail plausible phenotypes when considered together may reasonably be assumed to be reliable. However, the accuracy of inferred ancestral states for one or more characters may be questionable where combined reconstructions imply implausible phenotypes for a proportion of internal nodes. This criterion for assessing reconstructed ancestral states is applied here in evaluating inferences of ancestral limb morphology in the scincid lizard clade Lerista. Ancestral numbers of digits for the manus and pes inferred assuming the models that best fit the data entail ancestral digit configurations for many nodes that differ fundamentally from configurations observed among known species. However, when an alternative model is assumed for the pes, inferred ancestral digit configurations are invariably represented among observed phenotypes. This indicates that a suboptimal model for the pes (and not the model providing the best fit to the data) yields accurate ancestral state estimates. PMID- 20007168 TI - The effects of familiarity and social hierarchy on group membership decisions in a social fish. AB - Members of animal groups face a trade-off between the benefits of remaining with a familiar group and the potential benefits of dispersing into a new group. Here, we examined the group membership decisions of Neolamprologus pulcher, a group living cichlid. We found that subordinate helpers showed a preference for joining familiar groups, but when choosing between two unfamiliar groups, helpers did not preferentially join groups that maximized their social rank. Rather, helpers preferred groups containing larger, more dominant individuals, despite receiving significantly more aggression within these groups, possibly owing to increased protection from predation in such groups. These results suggest a complex decision process in N. pulcher when choosing among groups, dependent not only on familiarity but also on the social and life-history consequences of joining new groups. PMID- 20007169 TI - Does encephalization correlate with life history or metabolic rate in Carnivora? AB - A recent analysis of brain size evolution reconstructed the plesiomorphic brain body size allometry for the mammalian order Carnivora, providing an important reference frame for comparative analyses of encephalization (brain volume scaled to body mass). I performed phylogenetically corrected regressions to remove the effects of body mass, calculating correlations between residual values of encephalization with basal metabolic rate (BMR) and six life-history variables (gestation time, neonatal mass, weaning time, weaning mass, litter size, litters per year). No significant correlations were recovered between encephalization and any life-history variable or BMR, arguing against hypotheses relating encephalization to maternal energetic investment. However, after correcting for clade-specific adaptations, I recovered significant correlations for several variables, and further analysis revealed a conserved carnivoran reproductive strategy, linking degree of encephalization to the well-documented mammalian life history trade-off between neonatal mass and litter size. This strategy of fewer, larger offspring correlating with increased encephalization remains intact even after independent changes in encephalization allometries in the evolutionary history of this clade. PMID- 20007170 TI - Continuity of diabetes care is associated with avoidable hospitalizations: evidence from Taiwan's National Health Insurance scheme. AB - OBJECTIVE: Taiwan's health-care system allows patients to utilize specialty services without referrals by primary care providers. This discontinuity of care may lead to increases in future hospitalizations. This study aims to determine whether the discontinuity of care is associated with the risk of hospitalization. DESIGN: A secondary data analysis based on a claim data of a nationally representative random sample of diabetic patients in Taiwan. A usual provider continuity (UPC) index was developed-a ratio of the visits to the physician that subjects most usually see relevant to diabetes care to the total physician visits relevant to diabetes care-to investigate its association with the risk of hospitalization. SETTING: Taiwan's National Health Insurance scheme from 1997 through 2002. PARTICIPANTS: Totally 6476 diabetic patients. INTERVENTION(s) None. MAIN OUTCOME MEASURE(s) Diabetes-related short-term and long-term ambulatory care sensitive condition (ACSC) admissions. RESULTS: Patients with ACSC admissions had significantly lower UPC scores compared with those without ACSC admissions. Using a Cox regression model that controlling for age, sex, severity of diabetes and the number of total visits, patients with low to medium continuity of care (UPC <0.75) were found to be significantly associated with increased risk of hospitalization as compared with patients with high continuity of care, especially for long-term ACSC admissions (relative risk: 1.336 [1.019-1.751]). CONCLUSIONS: Higher continuity of care with usual providers for diabetic care is significantly associated with lower risk of future hospitalization for long-term diabetic complication admissions. To avoid future hospitalization, health policy stakeholders are encouraged to improve the continuity of care through strengthening the provider-patient relationships. PMID- 20007171 TI - The fluid dynamics of canine olfaction: unique nasal airflow patterns as an explanation of macrosmia. AB - The canine nasal cavity contains hundreds of millions of sensory neurons, located in the olfactory epithelium that lines convoluted nasal turbinates recessed in the rear of the nose. Traditional explanations for canine olfactory acuity, which include large sensory organ size and receptor gene repertoire, overlook the fluid dynamics of odorant transport during sniffing. But odorant transport to the sensory part of the nose is the first critical step in olfaction. Here we report new experimental data on canine sniffing and demonstrate allometric scaling of sniff frequency, inspiratory airflow rate and tidal volume with body mass. Next, a computational fluid dynamics simulation of airflow in an anatomically accurate three-dimensional model of the canine nasal cavity, reconstructed from high resolution magnetic resonance imaging scans, reveals that, during sniffing, spatially separate odour samples are acquired by each nostril that may be used for bilateral stimulus intensity comparison and odour source localization. Inside the nose, the computation shows that a unique nasal airflow pattern develops during sniffing, which is optimized for odorant transport to the olfactory part of the nose. These results contrast sharply with nasal airflow in the human. We propose that mammalian olfactory function and acuity may largely depend on odorant transport by nasal airflow patterns resulting from either the presence of a highly developed olfactory recess (in macrosmats such as the canine) or the lack of one (in microsmats including humans). PMID- 20007172 TI - Effects of disorder and motion in a radical pair magnetoreceptor. AB - A critical requirement in the proposed chemical model of the avian magnetic compass is that the molecules that play host to the magnetically sensitive radical pair intermediates must be immobilized and rotationally ordered within receptor cells. Rotational disorder would cause the anisotropic responses of differently oriented radical pairs within the same cell to interfere destructively, while rapid molecular rotation would tend to average the crucial anisotropic magnetic interactions and induce electron spin relaxation, reducing the sensitivity to the direction of the geomagnetic field. So far, experimental studies have been able to shed little light on the required degree of ordering and immobilization. To address this question, computer simulations have been performed on a collection of radical pairs undergoing restricted rigid-body rotation, coherent anisotropic spin evolution, electron spin relaxation and spin selective recombination reactions. It is shown that the ordering and motional constraints necessary for efficient magnetoreception can be simultaneously satisfied if the radical pairs are uniaxially ordered with a moderate order parameter and if their motional correlation time is longer than about a quarter of their lifetime. PMID- 20007173 TI - Computational limits to binary genes. AB - We analyse the trade-off between the speed with which a gene can propagate information, the noise of its output and its metabolic cost. Our main finding is that for any given level of metabolic cost there is an optimal trade-off between noise and processing speed. Any system with a non-vanishing leak expression rate is suboptimal, i.e. it will exhibit higher noise and/or slower speed than leak free systems with the same metabolic cost. We also show that there is an optimal Hill coefficient h which minimizes noise and metabolic cost at fixed speeds, and an optimal threshold K which minimizes noise. PMID- 20007174 TI - Adaptive winter survival strategies: defended energy levels in juvenile Atlantic salmon along a latitudinal gradient. AB - Current knowledge suggests that patterns of energy storage and depletion in animals are governed by behavioural trade-offs between risks associated with feeding and future energy demands. However, the length of adverse periods varies over geographical or climatic gradients. To explore the potential for genotypic sources of variation in behavioural trade-offs, we compared the winter energy depletion patterns among 13 wild populations of juvenile Atlantic salmon (Salmo salar L.) along a latitudinal gradient (58-70 degrees N) and performed common environment experiments of energy-state-dependent feeding. In the wild, winter lipid-depletion rates were lower for northern than for southern populations. The variation in spring lipid levels among the population was lower than autumn variation, with storage lipid levels clustered close to critical limits for survival. In semi-natural stream channels with natural food supply, hatchery reared fish originating from northern populations showed a positive scaling of feeding activity with decreasing energy levels, whereas southern populations did not. In conclusion, juvenile Atlantic salmon from northern populations defend their energy levels more strongly than fish from southern populations. Adaptive variation in feeding activity appears important for this difference. Thus, the present study shows a link between geographical patterns in storage energy trajectories and adaptive differences in state-dependent feeding motivation. PMID- 20007175 TI - Visual gaze control during peering flight manoeuvres in honeybees. AB - As animals travel through the environment, powerful reflexes help stabilize their gaze by actively maintaining head and eyes in a level orientation. Gaze stabilization reduces motion blur and prevents image rotations. It also assists in depth perception based on translational optic flow. Here we describe side-to side flight manoeuvres in honeybees and investigate how the bees' gaze is stabilized against rotations during these movements. We used high-speed video equipment to record flight paths and head movements in honeybees visiting a feeder. We show that during their approach, bees generate lateral movements with a median amplitude of about 20 mm. These movements occur with a frequency of up to 7 Hz and are generated by periodic roll movements of the thorax with amplitudes of up to + or - 60 degrees . During such thorax roll oscillations, the head is held close to horizontal, thereby minimizing rotational optic flow. By having bees fly through an oscillating, patterned drum, we show that head stabilization is based mainly on visual motion cues. Bees exposed to a continuously rotating drum, however, hold their head fixed at an oblique angle. This result shows that although gaze stabilization is driven by visual motion cues, it is limited by other mechanisms, such as the dorsal light response or gravity reception. PMID- 20007176 TI - Early origins of inflammation: microbial exposures in infancy predict lower levels of C-reactive protein in adulthood. AB - Ecological factors are important determinants of the development and function of anti-pathogen defences. Inflammation is a central part of innate immunity, but the developmental factors that shape the regulation of inflammation are not known. We test the hypothesis that microbial exposures in infancy are associated with high sensitivity C-reactive protein (CRP) in adulthood using prospective data from a birth cohort in the Philippines (n = 1461). Lower birth weight was associated with increased CRP, consistent with a role for inflammation in the widely documented inverse relationship between birth weight and adult cardiovascular diseases. In addition, higher levels of microbial exposure in infancy were associated with lower CRP. These associations were independent of socioeconomic status, measures of current body fat and other health behaviours. We conclude that measures of microbial exposure and nutrition during the pre natal and early post-natal periods are important predictors of CRP concentration in young adulthood. We speculate that the development of anti-inflammatory regulatory networks in response to early microbial exposure represents plasticity in the development of anti-pathogen defences, and that this process may help explain the low CRP concentrations in this population. PMID- 20007178 TI - Sex allocation in yellow-legged gulls (Larus michahellis) depends on nutritional constraints on production of large last eggs. AB - Male and female offspring can differ in their susceptibility to pre-natal (e.g. egg quality) and post-natal (e.g. sib-sib competition) conditions, and parents can therefore increase their individual fitness by adjusting these maternal effects according to offspring sex. In birds, egg mass and laying/hatching order are the main determinants of offspring viability, but these effects can act differently on each sex. In a previous study, relatively large last-laid (c-)eggs of yellow-legged gulls (Larus michahellis) were more likely to carry a female embryo. This suggests compensatory allocation of maternal resources to daughters from c-eggs, which suffer reduced viability. In the present study, we supplemented yellow-legged gulls with food during the laying period to experimentally test whether their nutritional conditions were responsible for the observed covariation between c-egg sex and mass. As predicted, food supplementation enhanced female c-eggs' mass more than that of male c-eggs. Thus, this experiment indicates that mothers strategically allocated their resources to c-eggs, possibly in order to compensate for the larger susceptibility of daughters to hatching (and laying) order. The results also suggested that mothers decided on resource allocation depending on the sex of already ovulated c-eggs, rather than ovulating ova of either sex depending on food availability. PMID- 20007179 TI - Poverty trap formed by the ecology of infectious diseases. AB - While most of the world has enjoyed exponential economic growth, more than one sixth of the world is today roughly as poor as their ancestors were many generations ago. Widely accepted general explanations for the persistence of such poverty have been elusive and are needed by the international development community. Building on a well-established model of human infectious diseases, we show how formally integrating simple economic and disease ecology models can naturally give rise to poverty traps, where initial economic and epidemiological conditions determine the long-term trajectory of the health and economic development of a society. This poverty trap may therefore be broken by improving health conditions of the population. More generally, we demonstrate that simple human ecological models can help explain broad patterns of modern economic organization. PMID- 20007177 TI - Grapheme-colour synaesthesia improves detection of embedded shapes, but without pre-attentive 'pop-out' of synaesthetic colour. AB - For people with synaesthesia letters and numbers may evoke experiences of colour. It has been previously demonstrated that these synaesthetes may be better at detecting a triangle made of 2s among a background of 5s if they perceive 5 and 2 as having different synaesthetic colours. However, other studies using this task (or tasks based on the same principle) have failed to replicate the effect or have suggested alternative explanations of the effect. In this study, we repeat the original study on a larger group of synaesthetes (n = 36) and include, for the first time, an assessment of their self-reported colour experiences. We show that synaesthetes do have a general advantage over controls on this task. However, many synaesthetes report no colour experiences at all during the task. Synaesthetes who do report colour typically experience around one third of the graphemes in the display as coloured. This is more consistent with theories of synaesthesia in which spatial attention needs to be deployed to graphemes for conscious colour experiences to emerge than the interpretation based on 'pop out'. PMID- 20007181 TI - Atractiellomycetes belonging to the 'rust' lineage (Pucciniomycotina) form mycorrhizae with terrestrial and epiphytic neotropical orchids. AB - Distinctive groups of fungi are involved in the diverse mycorrhizal associations of land plants. All previously known mycorrhiza-forming Basidiomycota associated with trees, ericads, liverworts or orchids are hosted in Agaricomycetes, Agaricomycotina. Here we demonstrate for the first time that Atractiellomycetes, members of the 'rust' lineage (Pucciniomycotina), are mycobionts of orchids. The mycobionts of 103 terrestrial and epiphytic orchid individuals, sampled in the tropical mountain rainforest of Southern Ecuador, were identified by sequencing the whole ITS1-5.8S-ITS2 region and part of 28S rDNA. Mycorrhizae of 13 orchid individuals were investigated by transmission electron microscopy. Simple septal pores and symplechosomes in the hyphal coils of mycorrhizae from four orchid individuals indicated members of Atractiellomycetes. Molecular phylogeny of sequences from mycobionts of 32 orchid individuals out of 103 samples confirmed Atractiellomycetes and the placement in Pucciniomycotina, previously known to comprise only parasitic and saprophytic fungi. Thus, our finding reveals these fungi, frequently associated to neotropical orchids, as the most basal living basidiomycetes involved in mycorrhizal associations of land plants. PMID- 20007180 TI - Retinal development and function in a 'blind' mole. AB - Animals adapted to dark ecotopes may experience selective pressure for retinal reduction. No previous studies have explicitly addressed the molecular basis of retinal development in any fossorial mammal. We studied retinal development and function in the Iberian mole Talpa occidentalis, which was presumed to be blind because of its permanently closed eyes. Prenatal retina development was relatively normal, with specification of all cell types and evidence of dorsoventral regionalization. Severe developmental defects occurred after birth, subsequent to lens abnormalities. 'Blind' Iberian moles had rods, cones and rod nuclear ultrastructure typical of diurnal mammals. DiI staining revealed only contralateral projections through the optic chiasm. Y-maze experiments demonstrated that moles retain a photoavoidance response. Over-representation of melanopsin-positive retinal ganglion cells that mediate photoperiodicity was observed. Hence, molecular pathways of eye development in Iberian moles retain the adaptive function of rod/cone primary vision and photoperiodicity, with no evidence that moles are likely to completely lose their eyes on an evolutionary time scale. PMID- 20007182 TI - A novel sex determination system in a close relative of the house mouse. AB - Therian mammals have an extremely conserved XX/XY sex determination system. A limited number of mammal species have, however, evolved to escape convention and present aberrant sex chromosome complements. In this study, we identified a new case of atypical sex determination in the African pygmy mouse Mus minutoides, a close evolutionary relative of the house mouse. The pygmy mouse is characterized by a very high proportion of XY females (74%, n = 27) from geographically widespread Southern and Eastern African populations. Sequencing of the high mobility group domain of the mammalian sex determining gene Sry, and karyological analyses using fluorescence in situ hybridization and G-banding data, suggest that the sex reversal is most probably not owing to a mutation of Sry, but rather to a chromosomal rearrangement on the X chromosome. In effect, two morphologically different X chromosomes were identified, one of which, designated X*, is invariably associated with sex-reversed females. The asterisk designates the still unknown mutation converting X*Y individuals into females. Although relatively still unexplored, such an atypical sex chromosome system offers a unique opportunity to unravel new genetic interactions involved in the initiation of sex determination in mammals. PMID- 20007183 TI - Biomechanics of the unique pterosaur pteroid. AB - Pterosaurs, flying reptiles from the Mesozoic, had wing membranes that were supported by their arm bones and a super-elongate fourth finger. Associated with the wing, pterosaurs also possessed a unique wrist bone--the pteroid--that functioned to support the forward part of the membrane in front of the leading edge, the propatagium. Pteroid shape varies across pterosaurs and reconstructions of its orientation vary (projecting anteriorly to the wing leading edge or medially, lying alongside it) and imply differences in the way that pterosaurs controlled their wings. Here we show, using biomechanical analysis and considerations of aerodynamic efficiency of a representative ornithocheirid pterosaur, that an anteriorly orientated pteroid is highly unlikely. Unless these pterosaurs only flew steadily and had very low body masses, their pteroids would have been likely to break if orientated anteriorly; the degree of movement required for a forward orientation would have introduced extreme membrane strains and required impractical tensioning in the propatagium membrane. This result can be generalized for other pterodactyloid pterosaurs because the resultant geometry of an anteriorly orientated pteroid would have reduced the aerodynamic performance of all wings and required the same impractical properties in the propatagium membrane. We demonstrate quantitatively that the more traditional reconstruction of a medially orientated pteroid was much more stable both structurally and aerodynamically, reflecting likely life position. PMID- 20007184 TI - The role of extinction in large-scale diversity-stability relationships. AB - More-diverse communities are thought to be ecologically stable because a greater number of ecological interactions among members allows for the increases in robustness and resilience. Diversity-stability relationships have mostly been studied on short ecological time scales but one study has identified such patterns over million-year time scales in reef communities. Here we propose and test a hypothesis for the mechanism of large-scale diversity-stability relationships in reefs. The extinction of community members destabilizes the community as a whole, unless there is sufficient diversity to buffer the community from the stochastic loss of members, thereby preventing collapse. If genera have high extinction rates, any variation in diversity among communities will result in a diversity-stability relationship. Conversely, in the absence of other mechanisms, the stability of low extinction communities is expected to be independent of diversity. We compare the extinction rates of six reef-building metazoan taxa to patterns of reef community stability and reef volume. We find that extinction of reef-builders occurs independent of reef volume, and that the strength of the diversity-stability relationship varies positively with extinction rate. PMID- 20007185 TI - Lost in translation or deliberate falsification? Genetic analyses reveal erroneous museum data for historic penguin specimens. AB - Historic museum specimens are increasingly used to answer a wide variety of questions in scientific research. Nevertheless, the scientific value of these specimens depends on the authenticity of the data associated with them. Here we use individual-based genetic analyses to demonstrate erroneous locality information for archive specimens from the late nineteenth century. Specifically, using 10 microsatellite markers, we analysed 350 contemporary and 43 historic yellow-eyed penguin (Megadyptes antipodes) specimens from New Zealand's South Island and sub-Antarctic regions. Factorial correspondence analysis and an assignment test strongly suggest that eight of the historic specimens purportedly of sub-Antarctic origin were in fact collected from the South Island. Interestingly, all eight specimens were obtained by the same collector, and all are currently held in the same museum collection. Further inspection of the specimen labels and evaluation of sub-Antarctic voyages did not reveal whether the erroneous data are caused by incorrect labelling or whether deliberate falsification was at play. This study highlights a promising extension to the well-known applications of assignment tests in molecular ecology, which can complement methods that are currently being applied for error detection in specimen data. Our results also serve as a warning to all who use archive specimens to invest time in the verification of collection information. PMID- 20007186 TI - Novel neogala-series glycosphingolipids with terminal mannose and glucose residues from Hirsutella rhossiliensis, an aureobasidin A-resistant ascomycete fungus. AB - Hirsutella rhossiliensis, a nematophagous fungus belonging to the Ascomycota, is resistant to aureobasidin A (AbA). In this fungus, the biosynthetic pathway leading to mannosylinositolphosphoceramides, which is inhibited by AbA, was not detected. Instead, this fungus contains neutral complex glycosphingolipids (GSLs) and monoglycosylceramides. Except for monoglycosylceramides, neutral GSLs share a neogala-series core structure, Galbeta1-6Galbeta1-Cer. Among the GSLs of H. rhossiliensis, three novel GSLs with terminal Man and Glc residues on the sugar chain were elucidated. We analyzed GSL structure using compositional sugar, fatty acid, and sphingoid analyses, methylation analysis, matrix-assisted laser desorption ionization time-of-flight/mass spectrometry (MALDI-TOF MS), and (1)H nuclear magnetic resonance spectroscopy (NMR). The following structures were determined: Manalpha1-3Galbeta1-6Galbeta1-6Galbeta1-Cer; Glcalpha1-2Galbeta1 6Galbeta1-6Galbeta1-Cer; and Manalpha1-3Galbeta1-6(Glcalpha1-4)Galbeta1-6Galbeta1 Cer. In the ceramides, the fatty acids were predominantly saturated h24:0-acids and the sphingoids were predominately t18:0- or t18:1-sphingoids. In contrast, the ceramides of Glcbeta1-Cer contained d18:2- and d19:2-sphingoids. These findings indicate the presence of a novel biosynthetic pathway of neogala-series GSLs in fungi. PMID- 20007187 TI - Extracorporeal cardiac mechanical stimulation: precordial thump and precordial percussion. AB - INTRODUCTION: External cardiac mechanical stimulation is one of the fastest resuscitative manoeuvres possible in the emergency setting. Precordial thump (PT), initially reported for treatment of atrio-ventricular block, has been subsequently described to cardiovert also ventricular tachycardia (VT) and fibrillation (VF). PT efficacy, mechanics and mechanisms remain poorly characterized. SOURCES OF DATA: Appropriate MESH and free terms were searched on PubMed, Embase and the Cochrane Library. Cross-referencing from articles and reviews, and forward search using SCOPUS and Google scholar have also been performed. Pre-set inclusion and exclusion criteria were applied to retrieved references on PT, which were then reviewed, summarized and interpreted. AREAS OF AGREEMENT: PT is not effective in treating VF, and of limited use for VT, although it has a very good safety profile (97% no changed/improved rhythm). If delivered, PT should be applied as early as possible after cardiac arrest, and cardio-pulmonary resuscitation (CPR) should begin with no delay if not effective. AREAS OF CONTROVERSY: A relatively large fraction of reported positive outcomes (both for PT and the less forceful but serially applied precordial percussion) in witnessed asystole should be considered when critically reviewing present CPR recommendations. In addition, mechanisms, energy requirements and timing are analysed and discussed. GROWING POINTS AND AREAS TIMELY FOR DEVELOPING RESEARCH: The 2005 ALS guidelines recommend PT delivery only by healthcare professionals trained in the technique. The use of training aids should therefore be explored, regardless of whether they are based on stand-alone devices or integrated within resuscitation mannequins. PMID- 20007188 TI - Socio-economic inequalities in childhood mortality in low- and middle-income countries: a review of the international evidence. AB - INTRODUCTION: In low- and middle-income countries (LMICs), the probability of dying in childhood is strongly related to the socio-economic position of the parents or household in which the child is born. This article reviews the evidence on the magnitude of socio-economic inequalities in childhood mortality within LMICs, discusses possible causes and highlights entry points for intervention. Sources of data Evidence on socio-economic inequalities in childhood mortality in LMICs is mostly based on data from household surveys and demographic surveillance sites. AREAS OF AGREEMENT: Childhood mortality is systematically and considerably higher among lower socio-economic groups within countries. Also most proximate mortality determinants, including malnutrition, exposure to infections, maternal characteristics and health care use show worse levels among more deprived groups. The magnitude of inequality varies between countries and over time, suggesting its amenability to intervention. Reducing inequalities in childhood mortality would substantially contribute to improving population health and reaching the Millennium Development Goals (MDGs). AREAS OF CONTROVERSY: The contribution of specific determinants, including national policies, to childhood mortality inequalities remains uncertain. What works to reduce these inequalities, in particular whether policies should be universal or targeted to the poor, is much debated. AREAS TIMELY FOR DEVELOPING RESEARCH: The increasing political attention for addressing health inequalities needs to be accompanied by more evidence on the contribution of specific determinants, and on ways to ensure that interventions reach lower socio-economic groups. PMID- 20007189 TI - Management of combined ACL-MCL tears: a systematic review. AB - INTRODUCTION: The optimal management for combined anterior cruciate ligament medial collateral ligament (ACL-MCL) injuries is controversial. SOURCES OF DATA: We performed a literature search using Medline, Cochrane and Google Scholar using the keywords: 'ACL' and 'MCL' in combination with 'surgery treatment', 'conservative treatment', 'surgery management', 'conservative management', 'surgical treatment' and 'surgical management'. We identified 23 published studies. AREAS OF AGREEMENT: Conservative and surgical management for combined ACL-MCL injuries resulted in different functional outcomes. The Coleman Methodology Score showed great heterogeneity in terms of study design, patient characteristics, management methods and outcome assessment and generally low methodological quality. AREAS OF CONTROVERSY: Given the heterogeneity in terms of treatment and results, we did not find a univocal trend over the years regarding MCL management (conservative or surgical). The use of several scoring systems did not allow us to compare outcomes in the different studies. GROWING POINTS: There is a need for a common validated scale for clinical measurements for ACL-MCL injuries, so as to allow easier and more reliable comparison of outcomes in different studies. To improve diagnostic certainty of combined ACL-MCL injuries, all patients should have imaging assessment (MR and stress-radiography) in addition to clinical examination. AREAS TIMELY FOR DEVELOPING RESEARCH: There is a need to perform appropriately powered randomized clinical trials of conservative and surgical treatment of combined ACL-MCL injuries, using standard diagnostic assessment, common and validated scoring system comparing reported outcomes and duration of follow-up more than 2 years. PMID- 20007190 TI - Successful management by provocative angiography and endovascular stent of Ureteroarterial fistula in a patient with a long-term indwelling ureteral stent. AB - We present the clinical course of a ureteroiliac arterial fistula in a patient who had been managed by ureteral stenting for 8 years for severe ureteral stricture after abdominoperineal resection with pelvic irradiation for advanced rectal cancer. A multidisciplinary team approach including provocative angiography and an endovascular stent saved the life of the patient. Ureteroarterial fistula is a rare complication of a long-term indwelling ureteral stent that is potentially fatal unless a prompt diagnosis and adequate therapy are provided. Heightened awareness and a high index of suspicion for this condition are required to make an early diagnosis. PMID- 20007191 TI - Quantitative investigation on the metabolism of 1,3-butadiene and of its oxidized metabolites in once-through perfused livers of mice and rats. AB - The industrial chemical 1,3-butadiene (BD) is a potent carcinogen in mice and a weak one in rats. This difference is generally related to species-specific burdens by the metabolites 1,2-epoxy-3-butene (EB), 1,2:3,4-diepoxybutane (DEB), and 3,4-epoxy-1,2-butanediol (EBD), which are all formed in the liver. Only limited data exist on BD metabolism in the rodent liver. Therefore, metabolism of BD, its epoxides, and the intermediate 3-butene-1,2-diol (B-diol) was studied in once-through perfused livers of male B6C3F1 mice and Sprague-Dawley rats. In BD perfusions, predominantly EB and B-diol were found (both species). DEB and EBD were additionally detected in mouse livers. Metabolism of BD showed saturation kinetics (both species). In EB perfusions, B-diol, EBD, and DEB were formed with B-diol being the major metabolite. Net formation of DEB was larger in mouse than in rat livers. In both species, hepatic clearance (Cl(H)) of EB was slightly smaller than the perfusion flow. In DEB perfusions, EBD was formed as a major metabolite. Cl(H) of DEB was 61% (mouse) and 73% (rat) of the perfusion flow. In the B-diol-perfused rat liver, EBD was formed as a minor metabolite. Cl(H) of B diol was 53% (mouse) and 34% (rat) of the perfusion flow. In EBD-perfused rat livers, Cl(H) of EBD represented only 22% of the perfusion flow. There is evidence for qualitative species differences with regard to the enzymes involved in BD metabolism. The first quantitative findings in whole livers showing intrahepatic first-pass metabolism of BD and EB metabolites will improve the risk estimation of BD. PMID- 20007196 TI - Applying ecological perspectives to adolescent sexual health in the United States: rhetoric or reality? AB - This study sought to determine the perspective taken toward understanding adolescent sexual risk behaviors and related biological outcomes (i.e. pregnancy, sexually transmitted diseases) since 1990. We content analyzed 324 abstracts representing observational research published between January 1990 and December 2007 for inclusion of ecological (environmental) factors, level of analysis, sample composition and type of behavioral and biological outcomes. A majority (95%) of studies included individual characteristics; half were void of any environmental factors. Of those including environmental factors, 27% included familial, 23% community, 13% relational and 3% societal factors. Most (80%) were positioned at the individual level of analysis. Samples were diverse (43%) and of mixed gender (71%). Biomarkers of sexually transmitted diseases (7.5%) or pregnancy outcomes (2%) were rare. Ecological inclusion was not related to year of publication. Despite the rhetoric highlighting, the importance of an ecological perspective in understanding adolescent sexual risk behavior, much published research, excludes environmental influences. PMID- 20007197 TI - Silymarin secretion and its elicitation by methyl jasmonate in cell cultures of Silybum marianum is mediated by phospholipase D-phosphatidic acid. AB - The flavonolignan silymarin is released to the extracellular medium of Silybum marianum cultures and its production can be stimulated by the elicitor methyljasmonate (MeJA). The sequence of the signalling processes leading to this response is unknown at present. It is reported in this work that MeJA increased the activity of the enzyme phospholipase D (PLD). Treatment with mastoparan (Mst), a PLD activity stimulator, also enhanced PLD and caused a substantial increase in silymarin production. The application of the product of PLD activity, phosphatidic acid (PA) promoted silymarin accumulation. Altering PLD activity by introducing in cultures n-butanol (nBuOH), which inhibits PA production by PLD, prevented silymarin elicitation by MeJA or Mst and also impeded its release in non-elicited cultures. Treatment with iso-, sec- or tert- butanol had no effect on silymarin production. The exogenous addition of PA reversed the inhibitory action of nBuOH, both in control and MeJA-treated cultures. These results suggest that the enzyme PLD and its product PA mediate silymarin secretion to the medium of S. marianum cultures. PMID- 20007198 TI - Cellulose microfibril orientation of Picea abies and its variability at the micron-level determined by Raman imaging. AB - The functional characteristics of plant cell walls depend on the composition of the cell wall polymers, as well as on their highly ordered architecture at scales from a few nanometres to several microns. Raman spectra of wood acquired with linear polarized laser light include information about polymer composition as well as the alignment of cellulose microfibrils with respect to the fibre axis (microfibril angle). By changing the laser polarization direction in 3 degrees steps, the dependency between cellulose and laser orientation direction was investigated. Orientation-dependent changes of band height ratios and spectra were described by quadratic linear regression and partial least square regressions, respectively. Using the models and regressions with high coefficients of determination (R(2) > 0.99) microfibril orientation was predicted in the S1 and S2 layers distinguished by the Raman imaging approach in cross sections of spruce normal, opposite, and compression wood. The determined microfibril angle (MFA) in the different S2 layers ranged from 0 degrees to 49.9 degrees and was in coincidence with X-ray diffraction determination. With the prerequisite of geometric sample and laser alignment, exact MFA prediction can complete the picture of the chemical cell wall design gained by the Raman imaging approach at the micron level in all plant tissues. PMID- 20007200 TI - The carnivorous bladderwort (Utricularia, Lentibulariaceae): a system inflates. AB - Carnivorous plants inhabit nutrient-poor environments, where prominent targets of prey capture are organic nitrogen and phosphorus. Some carnivorous plants also acquire carbon from their victims. A new report focusing on Utricularia, the bladderwort, demonstrates that carbon assimilated from photosynthesis is paradoxically secreted into the trapping environment, where it may help to support a mutualistic bacterial community. This bacterial community may also secrete allelochemicals that attract microcrustaceans which bear a strong overt resemblance to bladderwort traps. Furthermore, Utricularia and its sister genus Genlisea share anomalous molecular evolutionary features, such as highly increased rates of nucleotide substitution and dynamic evolution of genome size, from approximately 60-1500 megabases depending on the species or even population. A mechanistic hypothesis, based on the mutagenic action of reactive oxygen species (ROS) is proposed to underlie these phenomena, involving error-prone repair at the level of DNA bases and double-strand breaks. It is argued that these plants are prime candidates for further research on the complexities of plant physiology associated with carnivory, metagenomic surveys of trap microbial communities, novel plant nitrogen/nutrient utilization pathways, the ecology of prey attraction, whole-plant and trap comparative development, and, finally, evolution of the minimal angiosperm genome. PMID- 20007201 TI - Longitudinal data analysis for generalized linear models under participant-driven informative follow-up: an application in maternal health epidemiology. AB - It is common in longitudinal studies for scheduled visits to be accompanied by as needed visits due to medical events occurring between scheduled visits. If the timing of these as-needed visits is related to factors that are associated with the outcome but are not among the regression model covariates, naively including these as-needed visits in the model yields biased estimates. In this paper, the authors illustrate and discuss the key issues pertaining to inverse intensity rate ratio (IIRR)-weighted generalized estimating equations (GEE) methods in the context of a study of Kenyan mothers infected with human immunodeficiency virus type 1 (1999-2005). The authors estimated prevalences and prevalence ratios for morbid conditions affecting the women during a 1-year postpartum follow-up period. Of the 484 women under study, 62% had at least 1 as-needed visit. Use of a standard GEE model including both scheduled and unscheduled visits predicted a pneumonia prevalence of 2.9% (95% confidence interval: 2.3%, 3.5%), while use of the IIRR-weighted GEE predicted a prevalence of 1.5% (95% confidence interval: 1.2%, 1.8%). The estimate obtained using the IIRR-weighted GEE approach was compatible with estimates derived using scheduled visits only. These results highlight the importance of properly accounting for informative follow-up in these studies. PMID- 20007202 TI - Copy-years viremia as a measure of cumulative human immunodeficiency virus viral burden. AB - Plasma human immunodeficiency virus type 1 (HIV-1) viral load is a valuable tool for HIV research and clinical care but is often used in a noncumulative manner. The authors developed copy-years viremia as a measure of cumulative plasma HIV-1 viral load exposure among 297 HIV seroconverters from the Multicenter AIDS Cohort Study (1984-1996). Men were followed from seroconversion to incident acquired immunodeficiency syndrome (AIDS), death, or the beginning of the combination antiretroviral therapy era (January 1, 1996); the median duration of follow-up was 4.6 years (interquartile range (IQR), 2.7-6.5). The median viral load and level of copy-years viremia over 2,281 semiannual follow-up assessments were 29,628 copies/mL (IQR, 8,547-80,210) and 63,659 copies x years/mL (IQR, 15,935 180,341). A total of 127 men developed AIDS or died, and 170 survived AIDS-free and were censored on January 1, 1996, or lost to follow-up. Rank correlations between copy-years viremia and other measures of viral load were 0.56-0.87. Each log(10) increase in copy-years viremia was associated with a 1.70-fold increased hazard (95% confidence interval: 0.94, 3.07) of AIDS or death, independently of infection duration, age, race, CD4 cell count, set-point, peak viral load, or most recent viral load. Copy-years viremia, a novel measure of cumulative viral burden, may provide prognostic information beyond traditional single measures of viremia. PMID- 20007203 TI - Traumatic fracture of nitinol thermoreactive sternal clips. AB - Median sternotomy can be associated with significant morbidity, including non union, dehiscence and mediastinitis. The use of flexible thermoreactive sternal clips has been introduced recently as an alternative method of sternal closure and is advocated in patients at increased risk of sternal breakdown. It is associated with a decreased incidence of sternal complications as well as allowing faster sternal closure and easy removal on resternotomy. This report describes the case of a fractured thermoreactive clip following trauma, resulting in sternal dehiscence necessitating sternal rewiring. PMID- 20007204 TI - Granulation tissue formation at the bronchial stump is reduced after stapler closure in comparison to suture closure in dogs. AB - The aim of this study was to compare the morphology of the bronchial stump after lobectomy between mechanical stapler closure and manual suture closure. The effect of fibrin glue application on each method of closure was also observed. Right upper lobectomy was performed in beagles (n=31) using staplers (ST group) or sutures (SU group). In a separate experiment, fibrin glue was sprayed onto the stump after each respective method of closure. After one week, the stump region was examined macroscopically, and also by histology. chi(2)-Test and Mann-Whitney test were used for comparative analysis. The incidence of adhesion formation between the surrounding tissues was significantly reduced in the ST group in comparison to the SU group (22 vs. 80%, P=0.04). The thickness of granulation tissue over the stump was significantly reduced in the ST group in comparison to the SU group (0.8+/-0.2 vs. 2.5+/-0.3 mm, P<0.0001). Vessel density in the granulation tissue was also significantly reduced in the ST group in comparison to the SU group (6+/-2 vs. 16+/-2, P=0.003). Fibrin glue application after stapler closure significantly increased the incidence of adhesion formation, granulation tissue thickness, and vessel density in the granulation tissue over the stump. PMID- 20007205 TI - Infected aneurysm of the aortic arch with purulent pericarditis caused by Streptococcus pneumoniae. AB - A 76-year-old woman had a chest pain and high fever, and was admitted to the intensive care unit diagnosed as acute pericarditis. Enhanced CT-scan showed a 47 mm aneurysm in the aortic arch which seemed to be impending rupture and the part of the aorta looked like a pseudoaneurysm. Emergent total aortic arch replacement with a rifampicin-bonded Dacron graft was performed. Pericardial effusion was purulent and the aorta was infected with pus discharge in the aortic wall. There were some ulcerations on the surface of the luminal wall of the aorta. One of them was penetrating into the pericardial space causing a pseudoaneurysm. Both pericardial effusion and excised aortic wall were sent to culture study and resulted in positive for Streptococcus pneumoniae. The infection of the aorta, with erosion into the pericardial space, seemed to be the cause of purulent pericarditis. Antibiotic therapy was commenced immediately after surgery and continued for four weeks. Though she had neurological deficit after surgery, her infection was well controlled and there was no recurrence of infection 11 months after surgery. PMID- 20007206 TI - Painless amputation: history of a discovery that wasn't made. PMID- 20007214 TI - Health ministers from around the world make H1N1 top priority. PMID- 20007215 TI - Only 5% of the world's population is protected from second hand smoke by law, says WHO. PMID- 20007217 TI - Increased olfactory bulb volume due to treatment of chronic rhinosinusitis: neuroinflammation and adult neurogenesis. PMID- 20007218 TI - Autophagy induction reduces mutant ataxin-3 levels and toxicity in a mouse model of spinocerebellar ataxia type 3. AB - Spinocerebellar ataxia type 3 is a neurodegenerative disorder caused by the expansion of the polyglutamine repeat region within the ataxin-3 protein. The mutant protein forms intracellular aggregates in the brain. However, the cellular mechanisms causing toxicity are still poorly understood and there are currently no effective treatments. In this study we show that administration of a rapamycin ester (cell cycle inhibitor-779, temsirolimus) improves motor performance in a transgenic mouse model of spinocerebellar ataxia type 3. Temsirolimus inhibits mammalian target of rapamycin and hence upregulates protein degradation by autophagy. Temsirolimus reduces the number of aggregates seen in the brains of transgenic mice and decreases levels of cytosolic soluble mutant ataxin-3, while endogenous wild-type protein levels remain unaffected. Temsirolimus is designed for long-term use in patients and therefore represents a possible therapeutic strategy for the treatment of spinocerebellar ataxia type 3. Using this disease model and treatment paradigm, we employed a microarray approach to investigate transcriptional changes that might be important in the pathogenesis of spinocerebellar ataxia type 3. This identified ubiquitin specific peptidase-15, which showed expression changes at both the messenger ribonucleic acid and protein level. Ubiquitin specific peptidase-15 levels were also changed in mice expressing another mutant polyglutamine protein, huntingtin. In total we identified 16 transcripts that were decreased in transgenic ataxin-3 mice that were normalized following temsirolimus treatment. In this mouse model with relatively mild disease progression, the number of transcripts changed was low and the magnitude of these changes was small. However, the importance of these transcriptional alterations in the pathogenesis of spinocerebellar ataxia type 3 remains unclear. PMID- 20007220 TI - An 11 year old boy with chest pain and fever. PMID- 20007225 TI - Whitehall Study. Display of multiple factors. PMID- 20007226 TI - Tennis elbow. Injecting steroids is not good. PMID- 20007227 TI - Tennis elbow. Injections for all? PMID- 20007228 TI - Tennis elbow. Diagnose and treat the disease. PMID- 20007229 TI - Tennis elbow. There is no proved treatment. PMID- 20007230 TI - Quality in a poorer NHS. Gross and net are not the same. PMID- 20007231 TI - Dengue. Vector control is important. PMID- 20007232 TI - PSA and prostate cancer. More research is not needed. PMID- 20007234 TI - Antipsychotics and dementia. Mental Capacity Act 2005 should be invoked [corrected]. PMID- 20007235 TI - Trusts show some progress on tuberculosis but must coordinate their action, report says. PMID- 20007236 TI - Antipsychotics and dementia. Let's not waste more resources. PMID- 20007241 TI - Adult attachment insecurity and hippocampal cell density. AB - Recent findings associate attachment insecurity (assessed as levels of attachment anxiety and avoidance) with poor emotion regulation. In turn, emotion regulation has been shown to be associated with hippocampus (HC) functioning and structure. Clinical disorders such as depression and PTSD, which have been previously associated with attachment insecurity, are also known to be linked with reduced hippocampal cell density. This suggests that attachment insecurity may also be associated with reduced hippocampal cell density. We examined this hypothesis using T1 images of 22 healthy young adults. In line with our hypothesis, attachment avoidance was associated with bilateral HC reduction, whereas attachment anxiety was significantly related to reduced cell concentration in the left HC. The findings are compatible with a neurotoxical model of stress-induced cell reduction in the HC, providing further information on emotion regulation abilities among insecurely attached individuals. PMID- 20007242 TI - Self-involvement modulates the effective connectivity of the autobiographical memory network. AB - Neuroimaging studies have revealed a consistent overlap between brain regions involved in self-processing and those implicated in autobiographical memory. However, no study has directly tested how the degree of self-involvement with an event being remembered alters the neural circuitry engaged during memory retrieval. The present study compared hockey players' memories for game elements in which they were highly involved (e.g. scoring a goal) versus less involved (e.g. watching a goal from the bench). Specifically, we examined how the effective connectivity of a network of brain regions known to be involved in autobiographical memory retrieval varied based upon the players' level of self involvement with the remembered event. During remembering of high self involvement events, connections between the left hippocampus and medial prefrontal cortex were 'in synchrony' with connections between the medial prefrontal cortex and the right amygdala-hippocampal complex. By contrast, the hippocampal-prefrontal connection was 'out-of-sync' with the prefrontal-amygdala connection during retrieval of low self-involvement memories. This result is discussed in terms of two memory systems (one that is hippocampal-based and one that is amygdala-hippocampal-based) that may be involved to varying degrees depending upon the characteristics of a remembered event. PMID- 20007243 TI - Altered eosinophil profile in mice with ST6Gal-1 deficiency: an additional role for ST6Gal-1 generated by the P1 promoter in regulating allergic inflammation. AB - Cumulative evidence indicates that the sialyltransferase ST6Gal-1 and the sialyl glycans, which it constructs, are functionally pleiotropic. Expression of the ST6Gal-1 gene is mediated by six distinct promoter/regulatory regions, and we hypothesized that these promoters may be used differentially to produce ST6Gal-1 for different biologic purposes. To examine this hypothesis, we compared a mouse with a complete deficiency in ST6Gal-1 (Siat1 null) with another mouse that we have created previously with a disruption only in the P1 promoter (Siat1DeltaP1). We noted previously greater neutrophilic inflammation associated with ST6Gal-1 deficiency. Here, we report that ST6Gal-1-deficient mice also have significantly elevated eosinophilic responses. Upon i.p. thioglycollate elicitation, eosinophils accounted for over 20% of the total peritoneal inflammatory cell pool in ST6Gal-1-deficient animals, which was threefold greater than in corresponding wild-type animals. A principal feature of allergic respiratory inflammation is pulmonary eosinophilia, we evaluated the role of ST6Gal-1 in allergic lung inflammation. Using OVA and ABPA experimental models of allergic airways, we showed that ST6Gal-1 deficiency led to greater airway inflammation characterized by excessive airway eosinophilia. The severity of airway inflammation was similar between Siat1DeltaP1 and Siat1 null mice, indicating a role for P1-generated ST6Gal-1 in regulating eosinophilic inflammation. Colony-forming assays suggested greater IL-5-dependent eosinophil progenitor numbers in the marrow of ST6Gal-1 deficient animals. Moreover, allergen provocation of wild-type mice led to a significant reduction in P1-mediated ST6Gal-1 mRNA and accompanied decline in circulatory ST6Gal-1 levels. Taken together, the data implicate ST6Gal-1 as a participant in regulating not only Th1 but also Th2 responses, and ST6Gal-1 deficiency can lead to the development of more severe allergic inflammation with excessive eosinophil production. PMID- 20007244 TI - beta2-integrins in demyelinating disease: not adhering to the paradigm. AB - The beta(2)-integrins are a subfamily of integrins expressed on leukocytes that play an essential role in leukocyte trafficking, activation, and many other functions. Studies in EAE, the animal model for multiple sclerosis, show differential requirements for beta(2)-integrins in this disease model, ranging from critical in the case of LFA-1 (CD11a/CD18) to unimportant in the case of CD11d/CD18. Importantly, expression of beta(2)-integrins on T cell subsets provides some clues as to the function(s) these adhesion molecules play in disease development. For example, transferred EAE studies have shown that Mac-1 (CD11b/CD18) expression on alphabeta T cells is critical for disease development, and the absence of LFA-1 on Tregs in recipient mice results in exacerbated disease. In this review, we summarize recent findings regarding the role of beta(2)-integrins in demyelinating disease and new information about the role of beta(2)-integrins with respect to alterations in Treg numbers and function. In addition, we discuss the potential for targeting beta(2)-integrins in human demyelinating disease in light of the recent animal model studies. PMID- 20007245 TI - Dendritic cells transmit HIV-1 through human small intestinal mucosa. AB - To dissect the early events in the transmission of HIV-1 from mother to child, we investigated whether DCs participate in HIV-1 entry into human small intestinal mucosa. We isolated human MNLs from jejunal lamina propria and identified a subpopulation of CD11c(+)HLA-DR(+) MNLs that expressed DC-SIGN, CD83, CD86, CD206, and CCR7, indicating a DC phenotype. Jejunal DCs also expressed the HIV-1 receptor CD4 and coreceptors CCR5 and CXCR4 and in suspension rapidly took up cell-free HIV-1. HIV-1 inoculated onto the apical surface of explanted jejunum was transported by lamina propria DCs through the mucosa and transmitted in trans to blood and intestinal lymphocytes. These findings indicate that in addition to intestinal epithelial cells, which we showed previously transcytose infectious HIV-1 to indicator cells, intestinal DCs play an important role in transporting HIV-1 through the intestinal mucosa and the subsequent transmission to T cells. PMID- 20007247 TI - The murine IL-8 homologues KC, MIP-2, and LIX are found in endothelial cytoplasmic granules but not in Weibel-Palade bodies. AB - Rapid translocation of P-selectin from WPB to the surface of endothelial cells is crucial for early neutrophil recruitment to acute inflammatory lesions. Likewise, the chemokine CXCL8/IL-8 is sorted to WPB in human endothelial cells, but little is known about its functional importance in lack of a suitable animal model. Here, we explored the distribution of the functional IL-8 homologues CXCL1/KC, CXCL2/MIP-2, and CXCL5-6/LIX in resting and inflamed murine vessels by confocal microscopy and paired immunostaining with markers of WPB, discovering that these chemokines did not localize to WPB but displayed a granular pattern in a subset of vessels in healthy skin compatible with sorting to the type 2 endothelial compartment for regulated secretion. Moreover, all chemokines colocalized with VWF and P-selectin in platelets, suggesting that their storage in platelet alpha granules might represent an alternative source of rapidly available, neutrophil recruiting chemokines. In conclusion, WPB appear not to be involved in regulated secretion of chemokines in the mouse, and instead, the possible existence of type 2 granules and the role of platelets in rapid leukocyte adhesion deserve further attention. PMID- 20007246 TI - Proteome bioprofiles distinguish between M1 priming and activation states in human macrophages. AB - Macrophage activation is a dynamic process that results in diverse functional outcomes ranging from immunoregulation to inflammation. The proinflammatory, or M1, response is a complex, bimodal progression composed of a "prime," classically through IFN-gamma, and "trigger," such as LPS. To characterize the physiological response of M1 activation, a systems biology approach was applied to determine the intracellular proteome bioprofiles of IFN-gamma-and LPS-treated primary human macrophages. Our goal was to develop intracellular proteomic fingerprints to serve as novel correlates of macrophage priming and/or activation to augment the existing approaches of analyzing secreted cytokines and cell-surface protein expression. The majority of the proteome, approximately 78%, remained stable during activation, representing the core proteome. In contrast, three distinct patterns defined response proteomes: IFN-gamma-specific, LPS-specific, or IFN gamma- and LPS-shared or M1-specific. Although steady-state expression levels of proteins involved in energy metabolism and immune response were increased during priming and triggering, changes in protein and fatty acid metabolism, signaling, and transport pathways were most apparent. Unique proteomic fingerprints distinguish among IFN-gamma-specific, LPS-specific, or M1-specific activation states and provide a clear molecular, archeological profile to infer recent history of cells, as well as correlates for chronic macrophage activation in health and disease. PMID- 20007248 TI - A GMCSF-neuroantigen fusion protein is a potent tolerogen in experimental autoimmune encephalomyelitis (EAE) that is associated with efficient targeting of neuroantigen to APC. AB - Cytokine-NAg fusion proteins represent an emerging platform for specific targeting of self-antigen to particular APC subsets as a means to achieve antigen specific immunological tolerance. This study focused on cytokine-NAg fusion proteins that targeted NAg to myeloid APC. Fusion proteins contained GM-CSF or the soluble extracellular domain of M-CSF as the N-terminal domain and the encephalitogenic 69-87 peptide of MBP as the C-terminal domain. GMCSF-NAg and MCSF-NAg fusion proteins were approximately 1000-fold and 32-fold more potent than NAg in stimulating antigenic proliferation of MBP-specific T cells, respectively. The potentiated antigenic responses required cytokine-NAg covalent linkage and receptor-mediated uptake. That is, the respective cytokines did not potentiate antigenic responses when cytokine and NAg were added as separate molecules, and the potentiated responses were inhibited specifically by the respective free cytokine. Cytokine-dependent targeting of NAg was specific for particular subsets of APC. GMCSF-NAg and MCSF-NAg targeted NAg to DC and macrophages; conversely, IL4-NAg and IL2-NAg fusion proteins, respectively, induced an 1000-fold enhancement in NAg reactivity in the presence of B cell and T cell APC. GMCSF-NAg significantly attenuated severity of EAE when treatment was completed before encephalitogenic challenge or alternatively, when treatment was initiated after onset of EAE. MCSF-NAg also had significant tolerogenic activity, but GMCSF-NAg was substantially more efficacious as a tolerogen. Covalent GMCSF NAg linkage was required for prevention and treatment of EAE. In conclusion, GMCSF-NAg was highly effective for targeting NAg to myeloid APC and was a potent, antigen-specific tolerogen in EAE. PMID- 20007249 TI - Reduced T cell expansion by a superantigen as a result of impaired B cell development in mice deficient for the p85alpha regulatory subunit of PI3K. AB - PI3K plays crucial roles in the immune system. Mice deficient for p85alpha, a major regulatory subunit of class IA PI3K, show various defects and alterations in B cells, mast cells, macrophages, and DCs, and peripheral T cells are reportedly normal, at least in vitro. In normal mice, long-term exposure to a SAg, SEA, in vivo induced a high level of the protracted expansion of SEA reactive Vbeta3(+)CD4(+) T cells, whereas the same treatment induced T cell expansion in p85alpha-deficient mice but to a much lesser extent than in normal mice. However, mixed bone marrow chimera mice, which have normal and p85alpha deficient T and B cells, demonstrated equal responses of both T cells following stimulation with a SEA pump. In reciprocal cotransfer experiments of T and B cells from normal and p85alpha-deficient mice into Rag2-deficient mice, followed by SEA stimulation, p85alpha-deficient T cells revealed much higher proliferative capacity in the presence of normal B cells than did normal T cells with p85alpha deficient B cells. Histologically, a marked B cell reduction was observed in the follicles and MZ of the spleen, and DCs accumulated in the MZ. In addition, p85alpha-deficient B cells had a low level of MHC class II expression. Collectively, these data suggested that the PI3K p85alpha subunit alters the SAg presentation capacity of B cells and indirectly modulates the magnitude of the T cell response, which may affect the protection against SEA-containing bacteria. PMID- 20007250 TI - HVEM/LIGHT/BTLA/CD160 cosignaling pathways as targets for immune regulation. AB - Immunosuppression is currently the treatment of choice to attenuate the chronic deterioration of tissue function as a result of the effector mechanisms of the immunological response in transplant rejection and autoimmune diseases. However, global immunosuppression greatly increases the risk of acquiring life-threatening infections and is associated with organ toxicity when used long-term. Thus, alternative approaches that inhibit only the unwanted immune responses and preserve general immunity are highly desirable. The receptor/ligand pairs involved in the cross-talk between DC and T cells have been the focus of intense and exciting research during the last decade. The HVEM/LIGHT/BTLA/CD160 costimulatory/coinhibitory pathway has emerged as a potential target for the development of immune therapeutic interventions. Herein, we will summarize and discuss how blockade of the costimulatory HVEM/LIGHT interaction or agonist signaling through the inhibitory BTLA and CD160 receptors could contribute to the control of deleterious immune responses. PMID- 20007251 TI - ChiBE: interactive visualization and manipulation of BioPAX pathway models. AB - SUMMARY: Representing models of cellular processes or pathways in a graphically rich form facilitates interpretation of biological observations and generation of new hypotheses. Solving biological problems using large pathway datasets requires software that can combine data mapping, querying and visualization as well as providing access to diverse data resources on the Internet. ChiBE is an open source software application that features user-friendly multi-view display, navigation and manipulation of pathway models in BioPAX format. Pathway views are rendered in a feature-rich format, and may be laid out and edited with state-of the-art visualization methods, including compound or nested structures for visualizing cellular compartments and molecular complexes. Users can easily query and visualize pathways through an integrated Pathway Commons query tool and analyze molecular profiles in pathway context. AVAILABILITY: http://www.bilkent.edu.tr/%7Ebcbi/chibe.html. SUPPLEMENTARY INFORMATION: Supplementary data are available at Bioinformatics online. PMID- 20007252 TI - A novel method for accurate one-dimensional protein structure prediction based on fragment matching. AB - MOTIVATION: The precise prediction of one-dimensional (1D) protein structure as represented by the protein secondary structure and 1D string of discrete state of dihedral angles (i.e. Shape Strings) is a prerequisite for the successful prediction of three-dimensional (3D) structure as well as protein-protein interaction. We have developed a novel 1D structure prediction method, called Frag1D, based on a straightforward fragment matching algorithm and demonstrated its success in the prediction of three sets of 1D structural alphabets, i.e. the classical three-state secondary structure, three- and eight-state Shape Strings. RESULTS: By exploiting the vast protein sequence and protein structure data available, we have brought secondary-structure prediction closer to the expected theoretical limit. When tested by a leave-one-out cross validation on a non redundant set of PDB cutting at 30% sequence identity containing 5860 protein chains, the overall per-residue accuracy for secondary-structure prediction, i.e. Q3 is 82.9%. The overall per-residue accuracy for three- and eight-state Shape Strings are 85.1 and 71.5%, respectively. We have also benchmarked our program with the latest version of PSIPRED for secondary structure prediction and our program predicted 0.3% better in Q3 when tested on 2241 chains with the same training set. For Shape Strings, we compared our method with a recently published method with the same dataset and definition as used by that method. Our program predicted at 2.2% better in accuracy for three-state Shape Strings. By quantitatively investigating the effect of data base size on 1D structure prediction we show that the accuracy increases by approximately 1% with every doubling of the database size. PMID- 20007253 TI - PCIT: an R package for weighted gene co-expression networks based on partial correlation and information theory approaches. AB - SUMMARY: We make the PCIT algorithm, used for detecting meaningful gene-gene associations in co-expression networks, available as an R package. Automatic detection of a suitable parallel environment is used such that scripts are portable between parallel and non-parallel environments with no modification of the script. AVAILABILITY AND IMPLEMENTATION: Source code and binaries freely available (under GPL-3) for download via CRAN at http://cran.r project.org/package=PCIT, implemented in R and supported on Linux and MS Windows. PMID- 20007254 TI - ConceptGen: a gene set enrichment and gene set relation mapping tool. AB - MOTIVATION: The elucidation of biological concepts enriched with differentially expressed genes has become an integral part of the analysis and interpretation of genomic data. Of additional importance is the ability to explore networks of relationships among previously defined biological concepts from diverse information sources, and to explore results visually from multiple perspectives. Accomplishing these tasks requires a unified framework for agglomeration of data from various genomic resources, novel visualizations, and user functionality. RESULTS: We have developed ConceptGen, a web-based gene set enrichment and gene set relation mapping tool that is streamlined and simple to use. ConceptGen offers over 20,000 concepts comprising 14 different types of biological knowledge, including data not currently available in any other gene set enrichment or gene set relation mapping tool. We demonstrate the functionalities of ConceptGen using gene expression data modeling TGF-beta-induced epithelial mesenchymal transition and metabolomics data comparing metastatic versus localized prostate cancers. PMID- 20007255 TI - On the beta-binomial model for analysis of spectral count data in label-free tandem mass spectrometry-based proteomics. AB - MOTIVATION: Spectral count data generated from label-free tandem mass spectrometry-based proteomic experiments can be used to quantify protein's abundances reliably. Comparing spectral count data from different sample groups such as control and disease is an essential step in statistical analysis for the determination of altered protein level and biomarker discovery. The Fisher's exact test, the G-test, the t-test and the local-pooled-error technique (LPE) are commonly used for differential analysis of spectral count data. However, our initial experiments in two cancer studies show that the current methods are unable to declare at 95% confidence level a number of protein markers that have been judged to be differential on the basis of the biology of the disease and the spectral count numbers. A shortcoming of these tests is that they do not take into account within- and between-sample variations together. Hence, our aim is to improve upon existing techniques by incorporating both the within- and between sample variations. RESULT: We propose to use the beta-binomial distribution to test the significance of differential protein abundances expressed in spectral counts in label-free mass spectrometry-based proteomics. The beta-binomial test naturally normalizes for total sample count. Experimental results show that the beta-binomial test performs favorably in comparison with other methods on several datasets in terms of both true detection rate and false positive rate. In addition, it can be applied for experiments with one or more replicates, and for multiple condition comparisons. Finally, we have implemented a software package for parameter estimation of two beta-binomial models and the associated statistical tests. AVAILABILITY AND IMPLEMENTATION: A software package implemented in R is freely available for download at http://www.oncoproteomics.nl/. SUPPLEMENTARY INFORMATION: Supplementary data are available at Bioinformatics online. PMID- 20007256 TI - Dynamically weighted clustering with noise set. AB - MOTIVATION: Various clustering methods have been applied to microarray gene expression data for identifying genes with similar expression profiles. As the biological annotation data accumulated, more and more genes have been organized into functional categories. Functionally related genes may be regulated by common cellular signals, thus likely to be co-expressed. Consequently, utilizing the rapidly increasing functional annotation resources such as Gene Ontology (GO) to improve the performance of clustering methods is of great interest. On the opposite side of clustering, there are genes that have distinct expression profiles and do not co-express with other genes. Identification of these scattered genes could enhance the performance of clustering methods. RESULTS: We developed a new clustering algorithm, Dynamically Weighted Clustering with Noise set (DWCN), which makes use of gene annotation information and allows for a set of scattered genes, the noise set, to be left out of the main clusters. We tested the DWCN method and contrasted its results with those obtained using several common clustering techniques on a simulated dataset as well as on two public datasets: the Stanford yeast cell-cycle gene expression data, and a gene expression dataset for a group of genetically different yeast segregants. CONCLUSION: Our method produces clusters with more consistent functional annotations and more coherent expression patterns than existing clustering techniques. SUPPLEMENTARY INFORMATION: Supplementary data are available at Bioinformatics online. PMID- 20007257 TI - Clinical and epidemiological characteristics of children with Kawasaki disease in Turkey. AB - BACKGROUND: Kawasaki disease (KD) is the leading cause of acquired heart disease in childhood in the developed countries. The objective of this study is to describe the clinical and epidemiological characteristics of children with KD in Turkey. METHODS: The medical records of 24 patients treated for KD between January 1994 and June 2009 at Ankara University Medical School, Turkey were reviewed. RESULTS: The male-to-female ratio was 1.4 : 1. The median age at diagnosis was 2 years (range: 6.5 months to 11 years). Conjunctivitis and changes in the lips and oral cavity were seen in 21/24 (87.5%), cervical lymphadenopathy 17/24 (70.8%), polymorphous rash 16/24 (66.7%) and peripheral changes in 12/24 (50%). Coronary artery abnormality (CAA) was observed in 8/24 (33.3%) cases. CAA was seen in both the complete and incomplete groups with similar frequency (31.3% vs. 37.5%, respectively). CONCLUSIONS: KD must be kept in mind in the differential diagnosis of infants with prolonged fever. PMID- 20007258 TI - Herbal Medicine Containing Licorice May Be Contraindicated for a Patient with an HSD11B2 Mutation. AB - Licorice ingestion, as well as mutations in the HSD11B2 gene, inhibits 11beta hydroxysteroid dehydrogenase type 2 (11betaHSD2) enzyme activity, causing the syndrome of apparent mineral corticoid excess (AME). However, the combined effect of licorice ingestion and an HSD11B2 mutation has never been reported, until now. In this study, we demonstrated that licorice ingestion can produce overt hypertension in an individual without medical history of hypertension who is heterozygous for wild-type and mutant HSD11B2 genes. Our patient was a 51-year old female with serious hypertension who had been taking herbal medicine containing licorice for more than one year. She was clinically diagnosed as having licorice intoxication, because she did not present with hypertension after ceasing the herbal medicine. Molecular analysis showed that she carried a missense mutation, c.40C>T, in HSD11B2. In conclusion, licorice ingestion is an environmental risk factor for hypertension or AME state in patients with a mutation in HSD11B2. Carrying a mutation in HSD11B2 is, conversely, a genetic risk factor for licorice-induced hypertension or AME state. Herbal medicine containing licorice may, therefore, be contraindicated in patients with an HSD11B2 mutation. PMID- 20007259 TI - Brazilian green propolis inhibits inflammatory angiogenesis in a murine sponge model. AB - Angiogenesis and inflammation are persistent features of several pathological conditions. Propolis, a sticky material that honeybees collect from living plants, has been reported to have multiple biological effects including anti inflammatory and anti-neoplasic activities. Here, we investigated the effects of water extract of green propolis (WEP) on angiogenesis, inflammatory cell accumulation and endogenous production of cytokines in sponge implants of mice over a 14-day period. Blood vessel formation as assessed by hemoglobin content and by morphometric analysis of the implants was reduced by WEP (500 mg kg(-1) orally) compared to the untreated group. The levels of vascular endothelial growth factor (VEGF) increased progressively in the treated group but decreased after Day 10 in the control group. Accumulation of neutrophils and macrophages was determined by measuring myeloperoxidase (MPO) and N-acetyl-beta-(D) glucosaminidase (NAG) activities, respectively. Neutrophil accumulation was unaffected by propolis, but NAG activity was reduced by the treatment at Day 14. The levels TGF-beta1 intra-implant increased progressively in both groups but were higher (40%) at Day 14 in the control implants. The pro-inflammatory levels of TNF-alpha peaked at Day 7 in the control implants, and at Day 14 in the propolis-treated group. Our results indicate that the anti-inflammatory/anti angiogenic effects of propolis are associated with cytokine modulation. PMID- 20007260 TI - Ellagic Acid, the Active Compound of Phyllanthus urinaria, Exerts In Vivo Anti Angiogenic Effect and Inhibits MMP-2 Activity. AB - This study aimed to assess the potential anti-angiogenic mechanism of Phyllanthus urinaria (P. urinaria) and characterize the major compound in P. urinaria that exerts anti-angiogenic effect. The water extract of P. urinaria and Ellagic Acid were used to evaluate the anti-angiogenic effect in chorioallantoic membrane (CAM) in chicken embryo and human vascular endothelial cells (HUVECs). The matrix metalloproteinase-2 (MMP-2) activity was determined by gelatin zymography. The mRNA expressions of MMP-2, MMP-14 and tissue inhibitor of metalloproteinase-2 (TIMP-2) were analyzed by reverse transcription polymerase chain reaction (RT PCR). Level of MMP-2 proteins in conditioned medium or cytosol was determined by western blot analysis. We confirmed that P. urinaria's in vivo anti-angiogenic effect was associated with a reduction in MMP-2 activity. Ellagic acid, one of the major polyphenolic components as identified in P. urinaria by high performance liquid chromatography mass spectrometry (HPLC/MS), exhibited the same anti-angiogenic effect in vivo. Both P. urinaria and Ellagic Acid inhibited MMP-2 activity in HUVECs with unchanged mRNA level. The mRNA expression levels of MMP 14 and TIMP-2 were not altered either. Results from comparing the change of MMP-2 protein levels in conditioned medium and cytosol of HUVECs after the P. urinaria or Ellagic Acid treatment revealed an inhibitory effect on the secretion of MMP-2 protein. This study concluded that Ellagic Acid is the active compound in P. urinaria to exhibit anti-angiogenic activity and to inhibit the secretion of MMP 2 protein from HUVECs. PMID- 20007261 TI - Goal-directed acupuncture in sports-placebo or doping? AB - The modern pentathlon (MP), sports discipline including fencing, swimming, steeplechase and a cross-country run, requires a rapid change of central nervous and peripheral neuromuscular activity from one sport to another in order to achieve the best possible results. We describe the case where a top MP athlete was supported by a program of acupoint stimulation, which was directed to relieve the symptoms, preventing him from effective performance. Although the fact of acupoint stimulation was associated with improvement of his results, other factors like training effect, placebo and nonspecific physiological effects and their mechanisms in sports are discussed in a literature review. The popularity of complementary and alternative medicine methods among the athletes raises the question of their potential misuse as a doping in competitive sports. PMID- 20007262 TI - Water extract of ashwagandha leaves limits proliferation and migration, and induces differentiation in glioma cells. AB - Root extracts of Withania somnifera (Ashwagandha) are commonly used as a remedy for a variety of ailments and a general tonic for overall health and longevity in the Indian traditional medicine system, Ayurveda. We undertook a study to investigate the anti-proliferative and differentiation-inducing activities in the water extract of Ashwagandha leaves (ASH-WEX) by examining in glioma cells. Preliminary detection for phytochemicals was performed by thin-layer chromatography. Cytotoxicity was determined using trypan blue and MTT assays. Expression level of an hsp70 family protein (mortalin), glial cell differentiation marker [glial fibrillary acidic protein (GFAP)] and neural cell adhesion molecule (NCAM) were analyzed by immunocytochemistry and immunoblotting. Anti-migratory assay was also done using wound-scratch assay. Expression levels of mortalin, GFAP and NCAM showed changes, subsequent to the treatment with ASH WEX. The data support the existence of anti-proliferative, differentiation inducing and anti-migratory/anti-metastasis activities in ASH-WEX that could be used as potentially safe and complimentary therapy for glioma. PMID- 20007263 TI - Rotavirus infection activates dendritic cells from Peyer's patches in adult mice. AB - This study used an in vivo mouse model to analyze the response of dendritic cells (DCs) in Peyer's patches (PPs) within the first 48 h of infection with the wild type murine rotavirus EDIM (EDIM(wt)). After the infection, the absolute number of DCs was increased by 2-fold in the PPs without a modification of their relative percentage of the total cell number. Also, the DCs from PPs of infected mice showed a time-dependent migration to the subepithelial dome (SED) and an increase of the surface activation markers CD40, CD80, and CD86. This response was more evident at 48 h postinfection (p.i.) and depended on viral replication, since DCs from PPs of mice inoculated with UV-treated virus did not show this phenotype. As a result of the activation, the DCs showed an increase in the expression of mRNA for the proinflammatory cytokines interleukin-12/23p40 (IL 12/23p40), tumor necrosis factor alpha (TNF-alpha), and beta interferon (IFN beta), as well as for the regulatory cytokine IL-10. These results suggest that, a short time after rotavirus infection, the DCs from PPs play a critical role in controlling the infection and, at the same time, avoiding an excessive inflammatory immune response. PMID- 20007264 TI - The NS segment of an H5N1 highly pathogenic avian influenza virus (HPAIV) is sufficient to alter replication efficiency, cell tropism, and host range of an H7N1 HPAIV. AB - A reassortant avian influenza virus (designated FPV NS GD), carrying the NS segment of the highly pathogenic avian influenza virus (HPAIV) strain A/Goose/Guangdong/1/96 (GD; H5N1) in the genetic background of the HPAIV strain A/FPV/Rostock/34 (FPV; H7N1), was rescued by reverse genetics. Remarkably, in contrast to the recombinant wild-type FPV (rFPV), the reassortant virus was able to replicate more efficiently in different human cell lines and primary mouse epithelia cells without prior adaptation. Moreover, FPV NS GD caused disease and death in experimentally infected mice and was detected in mouse lungs; in contrast, rFPV was not able to replicate in mice effectively. These results indicated an altered host range and increased virulence. Furthermore FPV NS GD showed pronounced pathogenicity in chicken embryos. In an attempt to define the molecular basis for the apparent differences, we determined that NS1 proteins of the H5N1 and H7N1 strains bound the antiviral kinase PKR and the F2F3 domain of cleavage and polyadenylation specificity factor 30 (CPSF30) with comparable efficiencies in vitro. However, FPV NS GD infection resulted in (i) increased expression of NS1, (ii) faster and stronger PKR inhibition, and (iii) stronger beta interferon promoter inhibition than rFPV. Taken together, the results shed further light on the importance of the NS segment of an H5N1 strain for viral replication, molecular pathogenicity, and host range of HPAIVs and the possible consequences of a reassortment between naturally occurring H7 and H5 type HPAIVs. PMID- 20007265 TI - Fine mapping of pre-S sequence requirements for hepatitis B virus large envelope protein-mediated receptor interaction. AB - Previous studies showed that the N-terminal 75 amino acids of the pre-S1 domain of the hepatitis B virus (HBV) L protein are essential for HBV and hepatitis delta virus (HDV) infectivity. Consistently, synthetic lipopeptides encompassing this sequence or only parts of it efficiently block HBV and HDV infection, presumably through specific interference with a cellular receptor. Crucial for both virus infectivity and the inhibitory activity of the peptides are N-terminal myristoylation and a highly conserved motif within the N-terminal 48 amino acids. To refine the sequence requirements, we synthesized a series of HBV pre-S1 peptides containing deletions, point mutations, d-amino acid exchanges, or genotype-specific sequence permutations. Using the HepaRG cell line and a genotype D-derived virus, we determined the specific inhibitory activities of the peptides and found that (i) lipopeptides with an artificial consensus sequence inhibit HBV genotype D infection more potently than the corresponding genotype D peptides; (ii) point mutations, d-amino acid exchanges, or deletions introduced into the highly conserved part of the pre-S1 domain result in an almost complete loss of activity; and (iii) the flanking sequences comprising amino acids 2 to 8, 16 to 20, and, to a less pronounced extent, 34 to 48 gradually increase the inhibitory activity, while amino acids 21 to 33 behave indifferently. Taken together, our data suggest that HBV pre-S1-mediated receptor interference and, thus, HBV receptor recognition form a highly specific process. It requires an N terminal acyl moiety and a highly conserved sequence that is present in primate but not rodent or avian hepadnaviruses, indicating different entry pathways for the different family members. PMID- 20007266 TI - The prostate cancer-associated human retrovirus XMRV lacks direct transforming activity but can induce low rates of transformation in cultured cells. AB - The human retrovirus XMRV (xenotropic murine leukemia virus-related virus) is associated with prostate cancer, but a causal relationship has not been established. Here, we have used cultured fibroblast and epithelial cell lines to test the hypothesis that XMRV might have direct transforming activity but found only rare transformation events, suggestive of indirect transformation, even when the target cells expressed the human Xpr1 cell entry receptor for XMRV. Characterization of cells from three transformed foci showed that all were infected with and produced XMRV, and one produced a highly active transforming virus, presumably generated by recombination between XMRV and host cell nucleic acids. Given the sequence similarity of XMRV to mink cell focus-forming (MCF) viruses and the enhanced leukemogenic activity of the latter, we tested XMRV for related MCF-like cytopathic activities in cultured mink cells but found none. These results indicate that XMRV has no direct transforming activity but can activate endogenous oncogenes, resulting in cell transformation. As part of these experiments, we show that XMRV can infect and be produced at a high titer from human HT-1080 fibrosarcoma cells that express TRIM5alpha (Ref1), showing that XMRV is resistant to TRIM5alpha restriction. In addition, XMRV poorly infects NIH 3T3 cells expressing human Xpr1 but relatively efficiently infects BALB 3T3 cells expressing human Xpr1, showing that XMRV is a B-tropic virus and that its infectivity is regulated by the Fv1 mouse locus. PMID- 20007267 TI - Vaccination with an adenoviral vector that encodes and displays a retroviral antigen induces improved neutralizing antibody and CD4+ T-cell responses and confers enhanced protection. AB - We present a new type of adenoviral vector that both encodes and displays a vaccine antigen on the capsid, thus combining in itself gene-based and protein vaccination; this vector resulted in an improved vaccination outcome in the Friend virus (FV) model. For presentation of the envelope protein gp70 of Friend murine leukemia virus on the adenoviral capsid, gp70 was fused to the adenovirus capsid protein IX. When compared to vaccination with conventional FV Env- and Gag encoding adenoviral vectors, vaccination with the adenoviral vector that encodes and displays pIX-gp70 combined with an FV Gag-encoding vector resulted in significantly improved protection against systemic FV challenge infection, with highly controlled viral loads in plasma and spleen. This improved protection correlated with improved neutralizing antibody titers and stronger CD4(+) T-cell responses. Using a vector that displays gp70 without encoding it, we found that while the antigen display on the capsid alone was sufficient to induce high levels of binding antibodies, in vivo expression was necessary for the induction of neutralizing antibodies. This new type of adenovirus-based vaccine could be a valuable tool for vaccination. PMID- 20007268 TI - Importance of hydrogen bond contacts between the N protein and RNA genome of vesicular stomatitis virus in encapsidation and RNA synthesis. AB - Vesicular stomatitis virus (VSV) genomic RNA encapsidated by the nucleocapsid (N) protein is the template for transcription and replication by the viral polymerase. We analyzed the 2.9-A structure of the VSV N protein bound to RNA (T. J. Green, X. Zhang, G. W. Wertz, and M. Luo, Science 313:357-360, 2006) and identified amino acid residues with the potential to interact with RNA via hydrogen bonds. The contributions of these interactions to N protein function were investigated by individually substituting the residues with alanine and assaying the effect of these mutations on N protein expression, on the ability of the N protein to interact with the phosphoprotein (P), and on its ability to encapsidate RNA and generate templates that can support transcription and RNA replication. These studies identified individual amino acids critical for N protein function. Nine nucleotides are associated with each N monomer and contorted into two quasi-helices within the N protein RNA binding cavity. We found that N protein residues that formed hydrogen bond contacts with the nucleotides in quasi-helix 2 were critical to the encapsidation of RNA and the production of templates that can support RNA synthesis. Individual hydrogen bond interactions between the N protein and the nucleotides of quasi-helix 1 were not essential for ribonucleoprotein (RNP) template function. Residue R143 forms a hydrogen bond with nucleotide 9, the nucleotide that extends between N monomers. R143A mutant N protein failed to encapsidate RNA and to support RNA synthesis and suppressed wild-type N protein function. These studies show a direct correlation between viral RNA synthesis and N protein residues structurally positioned to interact with RNA. PMID- 20007269 TI - Effect of dimerizing domains and basic residues on in vitro and in vivo assembly of Mason-Pfizer monkey virus and human immunodeficiency virus. AB - Assembly of immature retroviral particles is a complex process involving interactions of several specific domains of the Gag polyprotein localized mainly within capsid protein (CA), spacer peptide (SP), and nucleocapsid protein (NC). In the present work we focus on the contribution of NC to the oligomerization of CA leading to assembly of Mason-Pfizer monkey virus (M-PMV) and HIV-1. Analyzing in vitro assembly of substitution and deletion mutants of DeltaProCANC, we identified a "spacer-like" sequence (NC(15)) at the M-PMV NC N terminus. This NC(15) domain is indispensable for the assembly and cannot be replaced with oligomerization domains of GCN4 or CREB proteins. Although the M-PMV NC(15) occupies a position analogous to that of the HIV-1 spacer peptide, it could not be replaced by the latter one. To induce the assembly, both M-PMV NC(15) and HIV 1 SP1 must be followed by a short peptide that is rich in basic residues. This region either can be specific, i.e., derived from the downstream NC sequence, or can be a nonspecific positively charged peptide. However, it cannot be replaced by heterologous interaction domains either from GCN4 or from CREB. In summary, we report here a novel M-PMV spacer-like domain that is functionally similar to other retroviral spacer peptides and contributes to the assembly of immature virus-like particles. PMID- 20007270 TI - Heterotypic humoral and cellular immune responses following Norwalk virus infection. AB - Norovirus immunity is poorly understood as the limited data available on protection after infection are often contradictory. In contrast to the more prominent GII noroviruses, GI norovirus infections are less frequent in outbreaks. The GI noroviruses display very complex patterns of heterotypic immune responses following infection, and many individuals are highly susceptible to reinfection. To study the immune responses and mechanisms of GI.1 persistence, we built structural models and recombinant virus-like particles (VLPs) of five GI strains: GI.1-1968, GI.1-2001, GI.2-1999, GI.3-1999, and GI.4-2000. Structural models of four GI genotype capsid P domain dimers suggested that intragenotype structural variation is limited, that the GI binding pocket is mostly preserved between genotypes, and that a conserved, surface-exposed epitope may allow for highly cross-reactive immune responses. GI VLPs bound to histo-blood group antigens (HBGAs) including fucose, Lewis, and A antigens. Volunteers infected with GI.1-1968 (n = 10) had significant increases between prechallenge and convalescent reactive IgG for all five GI VLPs measured by enzyme immunoassay. Potential cross-neutralization of GI VLPs was demonstrated by convalescent-phase serum cross-blockade of GI VLP-HBGA interaction. Although group responses were significant for all GI VLPs, each individual volunteer demonstrated a unique VLP blockade pattern. Further, peripheral blood mononuclear cells (PBMCs) were stimulated with each of the VLPs, and secretion of gamma interferon (IFN-gamma) was measured. As seen with blockade responses, IFN-gamma secretion responses differed by individual. Sixty percent responded to at least one GI VLP, with only two volunteers responding to GI.1 VLP. Importantly, four of five individuals with sufficient PBMCs for cross-reactivity studies responded more robustly to other GI VLPs. These data suggest that preexposure history and deceptive imprinting may complicate PBMC and B-cell immune responses in some GI.1-1968-challenged individuals and highlight a potential complication in the design of efficacious norovirus vaccines. PMID- 20007271 TI - Structure, receptor binding, and antigenicity of influenza virus hemagglutinins from the 1957 H2N2 pandemic. AB - The hemagglutinin (HA) envelope protein of influenza viruses mediates essential viral functions, including receptor binding and membrane fusion, and is the major viral antigen for antibody neutralization. The 1957 H2N2 subtype (Asian flu) was one of the three great influenza pandemics of the last century and caused 1 million deaths globally from 1957 to 1968. Three crystal structures of 1957 H2 HAs have been determined at 1.60 to 1.75 A resolutions to investigate the structural basis for their antigenicity and evolution from avian to human binding specificity that contributed to its introduction into the human population. These structures, which represent the highest resolutions yet recorded for a complete ectodomain of a glycosylated viral surface antigen, along with the results of glycan microarray binding analysis, suggest that a hydrophobicity switch at residue 226 and elongation of receptor-binding sites were both critical for avian H2 HA to acquire human receptor specificity. H2 influenza viruses continue to circulate in birds and pigs and, therefore, remain a substantial threat for transmission to humans. The H2 HA structure also reveals a highly conserved epitope that could be harnessed in the design of a broader and more universal influenza A virus vaccine. PMID- 20007272 TI - Z proteins of New World arenaviruses bind RIG-I and interfere with type I interferon induction. AB - The retinoic acid-inducible gene I product (RIG-I) is a cellular sensor of RNA virus infection that regulates the cellular beta interferon (IFN-beta) response. The nucleoproteins (NP) of arenaviruses are reported to antagonize the IFN response by inhibiting interferon regulatory factor 3 (IRF-3). Here, we demonstrate that the Z proteins of four New World (NW) arenaviruses, Guanarito virus (GTOV), Junin virus (JUNV), Machupo virus (MAVC), and Sabia virus (SABV), bind to RIG-I, resulting in downregulation of the IFN-beta response. We show that expression of the four NW arenavirus Z proteins inhibits IFN-beta mRNA induction in A549 cells in response to RNA bearing 5' phosphates (5'pppRNA). NW arenavirus Z proteins interact with RIG-I in coimmunoprecipitation studies and colocalize with RIG-I. Furthermore, expression of Z proteins interferes with the interaction between RIG-I and MAVS. Z expression also impedes the nuclear factor kappa light chain enhancer of activated B cells (NF-kappaB) and IRF-3 activation. Our results indicate that NW arenavirus Z proteins, but not Z protein of the Old World (OW) arenavirus lymphocytic choriomeningitis virus (LCMV) or Lassa virus, bind to RIG I and inhibit downstream activation of the RIG-I signaling pathway, preventing the transcriptional induction of IFN-beta. PMID- 20007273 TI - An N-terminal region of Lassa virus L protein plays a critical role in transcription but not replication of the virus genome. AB - The central domain of the 200-kDa Lassa virus L protein is a putative RNA dependent RNA polymerase. N- and C-terminal domains may harbor enzymatic functions important for viral mRNA synthesis, including capping enzymes or cap snatching endoribonucleases. In the present study, we have employed a large-scale mutagenesis approach to map functionally relevant residues in these regions. The main targets were acidic (Asp and Glu) and basic residues (Lys and Arg) known to form catalytic and binding sites of capping enzymes and endoribonucleases. A total of 149 different mutants were generated and tested in the Lassa virus replicon system. Nearly 25% of evolutionarily highly conserved acidic and basic side chains were dispensable for function of L protein in the replicon context. The vast majority of the remaining mutants had defects in both transcription and replication. Seven residues (Asp-89, Glu-102, Asp-119, Lys-122, Asp-129, Glu-180, and Arg-185) were selectively important for mRNA synthesis. The phenotype was particularly pronounced for Asp-89, Glu-102, and Asp-129, which were indispensable for transcription but could be replaced by a variety of amino acid residues without affecting genome replication. Bioinformatics disclosed the remote similarity of this region to type IIs endonucleases. The mutagenesis was complemented by experiments with the RNA polymerase II inhibitor alpha-amanitin, demonstrating dependence of viral transcription from the cellular mRNA pool. In conclusion, this paper describes an N-terminal region in L protein being important for mRNA, but not genome synthesis. Bioinformatics and cell biological experiments lend support to the hypothesis that this region could be part of a cap-snatching enzyme. PMID- 20007274 TI - During lytic infections, herpes simplex virus type 1 DNA is in complexes with the properties of unstable nucleosomes. AB - The genomes of herpes simplex virus type 1 (HSV-1) are regularly chromatinized during latency such that their digestion with micrococcal nuclease (MCN) releases nucleosome-sized DNA fragments. In lytically infected cells, in contrast, MCN releases HSV-1 DNA in primarily heterogeneously sized fragments. Consistently, only a small percentage of this HSV-1 DNA coimmunoprecipitates with histones. Most current models propose that histones associate with HSV-1 DNA during lytic infections at low occupancy. However, histone modification or occupation is also proposed to regulate HSV-1 transcription. It remains unclear how the histones associated with a small percentage of HSV-1 DNA may regulate transcription globally. Moreover, the physical properties of the complexes containing histones and HSV-1 DNA are unknown. We evaluated the HSV-1 DNA-containing complexes at 5 h after (lytic) infection by biochemical fractionations. Nuclear HSV-1 DNA did not fractionate as protein-free HSV-1 DNA but as DNA in cellular nucleosomes. Moreover, MCN released HSV-1 DNA in complexes that fractionate as cellular mono- and dinucleosomes by centrifugation followed by sucrose gradients and size exclusion chromatography. The HSV-1 DNA in such complexes was protected to heterogeneous sizes and was more accessible to MCN than DNA in most cellular chromatin. Using a modified MCN digestion to trap unstable digestion intermediates, HSV-1 DNA was quantitatively recovered in discrete mono- to polynucleosome sizes in complexes fractionating as cellular mono- to polynucleosomes. The HSV-1 DNAs in complexes fractionating as mono- to dinucleosomes were stabilized by cross-linking. Therefore, most HSV-1 DNA forms particularly unstable nucleosome-like complexes at 5 h of lytic infection. PMID- 20007275 TI - Crystal structure of an Aquabirnavirus particle: insights into antigenic diversity and virulence determinism. AB - Infectious pancreatic necrosis virus (IPNV), a pathogen of salmon and trout, imposes a severe toll on the aquaculture and sea farming industries. IPNV belongs to the Aquabirnavirus genus in the Birnaviridae family of bisegmented double stranded RNA viruses. The virions are nonenveloped with a T=13l icosahedral capsid made by the coat protein VP2, the three-dimensional (3D) organization of which is known in detail for the family prototype, the infectious bursal disease virus (IBDV) of poultry. A salient feature of the birnavirus architecture is the presence of 260 trimeric spikes formed by VP2, projecting radially from the capsid. The spikes carry the principal antigenic sites as well as virulence and cell adaptation determinants. We report here the 3.4-A resolution crystal structure of a subviral particle (SVP) of IPNV, containing 20 VP2 trimers organized with icosahedral symmetry. We show that, as expected, the SVPs have a very similar organization to the IBDV counterparts, with VP2 exhibiting the same overall 3D fold. However, the spikes are significantly different, displaying a more compact organization with tighter packing about the molecular 3-fold axis. Amino acids controlling virulence and cell culture adaptation cluster differently at the top of the spike, i.e., in a central bowl in IBDV and at the periphery in IPNV. In contrast, the spike base features an exposed groove, conserved across birnavirus genera, which contains an integrin-binding motif. Thus, in addition to revealing the viral antigenic determinants, the structure suggests that birnaviruses interact with different receptors for attachment and for cell internalization during entry. PMID- 20007276 TI - Multiple diverse circoviruses infect farm animals and are commonly found in human and chimpanzee feces. AB - Circoviruses are known to infect birds and pigs and can cause a wide range of severe symptoms with significant economic impact. Using viral metagenomics, we identified circovirus-like DNA sequences and characterized 15 circular viral DNA genomes in stool samples from humans in Pakistan, Nigeria, Tunisia, and the United States and from wild chimpanzees. Distinct genomic features and phylogenetic analysis indicate that some viral genomes were part of a previously unrecognized genus in the Circoviridae family we tentatively named "Cyclovirus" whose genetic diversity is comparable to that of all the known species in the Circovirus genus. Circoviridae detection in the stools of U.S. adults was limited to porcine circoviruses which were also found in most U.S. pork products. To determine whether the divergent cycloviruses found in non-U.S. human stools were of dietary origin, we genetically compared them to the cycloviruses in muscle tissue samples of commonly eaten farm animals in Pakistan and Nigeria. Limited genetic overlap between cycloviruses in human stool samples and local cow, goat, sheep, camel, and chicken meat samples indicated that the majority of the 25 Cyclovirus species identified might be human viruses. We show that the genetic diversity of small circular DNA viral genomes in various mammals, including humans, is significantly larger than previously recognized, and frequent exposure through meat consumption and contact with animal or human feces provides ample opportunities for cyclovirus transmission. Determining the role of cycloviruses, found in 7 to 17% of non-U.S. human stools and 3 to 55% of non-U.S. meat samples tested, in both human and animal diseases is now facilitated by knowledge of their genomes. PMID- 20007277 TI - Genetic analysis of the carboxy-terminal region of the hepatitis C virus core protein. AB - Hepatitis C virus (HCV) is a liver-tropic pathogen with severe health consequences for infected individuals. Chronic HCV infection can progress to cirrhosis and hepatocellular carcinoma and is a leading indicator for liver transplantation. The HCV core protein is an essential component of the infectious virus particle, but many aspects of its role remain undefined. The C-terminal region of the core protein acts as a signal sequence for the E1 glycoprotein and undergoes dual processing events during infectious virus assembly. The exact C terminus of the mature, virion-associated core protein is not known. Here, we performed genetic analyses to map the essential determinants of the HCV core C terminal region, as well as to define the minimal length of the protein that can function for infectious virus production in trans. PMID- 20007279 TI - Influenza C virus NS1 protein upregulates the splicing of viral mRNAs. AB - Pre-mRNAs of the influenza A virus M and NS genes are poorly spliced in virus infected cells. By contrast, in influenza C virus-infected cells, the predominant transcript from the M gene is spliced mRNA. The present study was performed to investigate the mechanism by which influenza C virus M gene-specific mRNA (M mRNA) is readily spliced. The ratio of M1 encoded by a spliced M mRNA to CM2 encoded by an unspliced M mRNA in influenza C virus-infected cells was about 10 times larger than that in M gene-transfected cells, suggesting that a viral protein(s) other than M gene translational products facilitates viral mRNA splicing. RNase protection assays showed that the splicing of M mRNA in infected cells was much higher than that in M gene-transfected cells. The unspliced and spliced mRNAs of the influenza C virus NS gene encode two nonstructural (NS) proteins, NS1(C/NS1) and NS2(C/NS2), respectively. The introduction of premature translational termination into the NS gene, which blocked the synthesis of the C/NS1 and C/NS2 proteins, drastically reduced the splicing of NS mRNA, raising the possibility that C/NS1 or C/NS2 enhances viral mRNA splicing. The splicing of influenza C virus M mRNA was increased by coexpression of C/NS1, whereas it was reduced by coexpression of the influenza A virus NS1 protein (A/NS1). The splicing of influenza A virus M mRNA was also increased by coexpression of C/NS1, though it was inhibited by that of A/NS1. These results suggest that influenza C virus NS1, but not A/NS1, can upregulate viral mRNA splicing. PMID- 20007278 TI - Dynamics of coronavirus replication-transcription complexes. AB - Coronaviruses induce in infected cells the formation of double-membrane vesicles (DMVs) in which the replication-transcription complexes (RTCs) are anchored. To study the dynamics of these coronavirus replicative structures, we generated recombinant murine hepatitis coronaviruses that express tagged versions of the nonstructural protein nsp2. We demonstrated by using immunofluorescence assays and electron microscopy that this protein is recruited to the DMV-anchored RTCs, for which its C terminus is essential. Live-cell imaging of infected cells demonstrated that small nsp2-positive structures move through the cytoplasm in a microtubule-dependent manner. In contrast, large fluorescent structures are rather immobile. Microtubule-mediated transport of DMVs, however, is not required for efficient replication. Biochemical analyses indicated that the nsp2 protein is associated with the cytoplasmic side of the DMVs. Yet, no recovery of fluorescence was observed when (part of) the nsp2-positive foci were bleached. This result was confirmed by the observation that preexisting RTCs did not exchange fluorescence after fusion of cells expressing either a green or a red fluorescent nsp2. Apparently, nsp2, once recruited to the RTCs, is not exchanged with nsp2 present in the cytoplasm or at other DMVs. Our data show a remarkable resemblance to results obtained recently by others with hepatitis C virus. The observations point to intriguing and as yet unrecognized similarities between the RTC dynamics of different plus-strand RNA viruses. PMID- 20007280 TI - Insertion mutations in herpes simplex virus 1 glycoprotein H reduce cell surface expression, slow the rate of cell fusion, or abrogate functions in cell fusion and viral entry. AB - Of the four required herpes simplex virus (HSV) entry glycoproteins, the precise role of gH-gL in fusion remains the most elusive. The heterodimer gH-gL has been proposed to mediate hemifusion after the interaction of another required glycoprotein, gD, with a receptor. To identify functional domains of HSV-1 gH, we generated 22 randomized linker-insertion mutants. Analyses of 22 gH mutants revealed that gH is relatively tolerant of insertion mutations, as 15 of 22 mutants permitted normal processing and transport of gH-gL to the cell surface. gH mutants that were not expressed well at the cell surface did not function in fusion or viral entry. The screening of gH mutants for function revealed the following: (i) for wild-type gH and some gH mutants, fusion with nectin-1 expressing target cells occurred more rapidly than with herpesvirus entry mediator (HVEM)-expressing target cells; (ii) some gH mutants reduced the rate of cell fusion without abrogating fusion completely, indicating that gH may play a role in governing the kinetics of fusion and may be responsible for a rate limiting first stage in HSV-1 fusion; and (iii) only one gH mutant, located within the short cytoplasmic tail, completely abrogated function, indicating that the gH cytoplasmic tail is crucial for cell fusion and viral infectivity. PMID- 20007281 TI - A rotavirus spike protein conformational intermediate binds lipid bilayers. AB - During rotavirus entry, a virion penetrates a host cell membrane, sheds its outer capsid proteins, and releases a transcriptionally active subviral particle into the cytoplasm. VP5, the rotavirus protein believed to interact with the membrane bilayer, is a tryptic cleavage product of the outer capsid spike protein, VP4. When a rotavirus particle uncoats, VP5 folds back, in a rearrangement that resembles the fusogenic conformational changes in enveloped-virus fusion proteins. We present direct experimental evidence that this rearrangement leads to membrane binding. VP5 does not associate with liposomes when mounted as part of the trypsin-primed spikes on intact virions, nor does it do so after it has folded back into a stably trimeric, low-energy state. But it does bind liposomes when they are added to virions before uncoating, and VP5 rearrangement is then triggered by addition of EDTA. The presence of liposomes during the rearrangement enhances the otherwise inefficient VP5 conformational change. A VP5 fragment, VP5CT, produced from monomeric recombinant VP4 by successive treatments with chymotrypsin and trypsin, also binds liposomes only when the proteolysis proceeds in their presence. A monoclonal antibody that neutralizes infectivity by blocking a postattachment entry event also blocks VP5 liposome association. We propose that VP5 binds lipid bilayers in an intermediate conformational state, analogous to the extended intermediate conformation of enveloped-virus fusion proteins. PMID- 20007283 TI - A single tyrosine in the severe acute respiratory syndrome coronavirus membrane protein cytoplasmic tail is important for efficient interaction with spike protein. AB - Severe acute respiratory syndrome coronavirus (SARS-CoV) encodes 3 major envelope proteins: spike (S), membrane (M), and envelope (E). Previous work identified a dibasic endoplasmic reticulum retrieval signal in the cytoplasmic tail of SARS CoV S that promotes efficient interaction with SARS-CoV M. The dibasic signal was shown to be important for concentrating S near the virus assembly site rather than for direct interaction with M. Here, we investigated the sequence requirements of the SARS-CoV M protein that are necessary for interaction with SARS-CoV S. The SARS-CoV M tail was shown to be necessary for S localization in the Golgi region when the proteins were exogenously coexpressed in cells. This was specific, since SARS-CoV M did not retain an unrelated glycoprotein in the Golgi. Importantly, we found that an essential tyrosine residue in the SARS-CoV M cytoplasmic tail, Y(195), was important for S-M interaction. When Y(195) was mutated to alanine, M(Y195A) no longer retained S intracellularly at the Golgi. Unlike wild-type M, M(Y195A) did not reduce the amount of SARS-CoV S carbohydrate processing or surface levels when the two proteins were coexpressed. Mutating Y(195) also disrupted SARS-CoV S-M interaction in vitro. These results suggest that Y(195) is necessary for efficient SARS-CoV S-M interaction and, thus, has a significant involvement in assembly of infectious virus. PMID- 20007282 TI - Nucleolin associates with the human cytomegalovirus DNA polymerase accessory subunit UL44 and is necessary for efficient viral replication. AB - In the eukaryotic cell, DNA replication entails the interaction of multiple proteins with the DNA polymerase processivity factor PCNA. As the structure of the presumptive human cytomegalovirus (HCMV) DNA polymerase processivity factor UL44 is highly homologous to that of PCNA, we hypothesized that UL44 also interacts with numerous proteins. To investigate this possibility, recombinant HCMV expressing FLAG-tagged UL44 was generated and used to immunoprecipitate UL44 and associated proteins from infected cell lysates. Unexpectedly, nucleolin, a major protein component of the nucleolus, was identified among these proteins by mass spectrometry and Western blotting. The association of nucleolin and UL44 in infected cell lysate was confirmed by reciprocal coimmunoprecipitation in the presence and absence of nuclease. Western blotting and immunofluorescence assays demonstrated that the level of nucleolin increases during infection and that nucleolin becomes distributed throughout the nucleus. Furthermore, the colocalization of nucleolin and UL44 in infected cell nuclei was observed by immunofluorescence assays. Assays of HCMV-infected cells treated with small interfering RNA (siRNA) targeting nucleolin mRNA indicated that nucleolin was required for efficient virus production, viral DNA synthesis, and the expression of a late viral protein, with a correlation between the efficacy of knockdown and the effect on virus replication. In contrast, the level of neither global protein synthesis nor the replication of an unrelated virus (reovirus) was reduced in siRNA-treated cells. Taken together, our results indicate an association of nucleolin and UL44 in HCMV-infected cells and a role for nucleolin in viral DNA synthesis. PMID- 20007284 TI - The murine coronavirus nucleocapsid gene is a determinant of virulence. AB - The murine coronavirus, mouse hepatitis virus (MHV) strain A59, causes acute encephalitis and chronic demyelinating disease as well as hepatitis in mice. The JHM strain (also called MHV-4 or JHM.SD) causes fatal encephalitis and only minimal hepatitis. Previous analysis of chimeric recombinant MHVs in which the spike gene, encoding the protein that mediates viral entry and cell-to-cell fusion, was exchanged between JHM and A59 showed that the spike plays a major role in determining organ tropism and neurovirulence but that other genes also play important roles in pathogenic outcome. Here, we have investigated the role of the nucleocapsid protein in MHV-induced disease. The multifunctional nucleocapsid protein is complexed with the genomic RNA, interacts with the viral membrane protein during virion assembly, and plays an import role in enhancing the efficiency of transcription. A pair of chimeric recombinant viruses in which the nucleocapsid gene was exchanged between JHM and A59 was selected and compared to wild-type parental strains in terms of virulence. Importantly, expression of the JHM nucleocapsid in the context of the A59 genome conferred increased mortality and spread of viral antigen in the mouse central nervous system compared to the parental A59 strain, while having little effect on the induction of hepatitis. While the JHM nucleocapsid did not appear to enhance neuron-to neuron spread in primary neuronal cultures, the increased neurovirulence it conferred may be due in part to the induction of a less robust T-cell response than that induced by strain A59. PMID- 20007285 TI - Rosiglitazone alleviates the persistent fibrotic phenotype of lesional skin scleroderma fibroblasts. AB - OBJECTIVE: The transcription factor peroxisome proliferator-activated receptor (PPAR)-gamma plays an important role in controlling cell differentiation. The aim of the present study was to examine whether PPAR-gamma expression was reduced in skin scleroderma fibroblasts and whether PPAR-gamma agonists could suppress the persistent fibrotic phenotype of skin scleroderma fibroblasts. METHODS: Dermal fibroblasts were isolated from site-, age- and sex-matched healthy individuals and lesional areas of individuals with dcSSc. Western blot and collagen gel contraction analyses were used to detect protein expression in the presence or absence of the PPAR-gamma agonist rosiglitazone. RESULTS: PPAR-gamma expression was reduced in dcSSc fibroblasts. The PPAR-gamma agonist rosiglitazone alleviated the persistent fibrotic phenotype of dcSSc fibroblasts. CONCLUSION: Rosiglitazone may alleviate the extent of fibrosis in dcSSc. PMID- 20007287 TI - Positron emission tomography/computed tomography: a clinical tool for evaluation of enthesitis in patients with spondyloarthritides. AB - OBJECTIVE: To evaluate the accuracy of PET/CT using [(18)F]fluorodeoxyglucose (FDG) in determining the presence of enthesitis in patients with SpAs. METHODS: Results of PET/CT scans of eight patients with SpA and seven patients with RA were retrospectively examined, with specific focus on five joints and three entheses. Volume fixation values are expressed as standardized uptake values (SUVs). Data from 20 patients with non-rheumatic (NR) diseases and 20 healthy subjects were also examined if non-specific, false positive findings were possible. We evaluated the clinical utility of PET/CT examinations in SpA, compared with MRI and Ga scintigraphy. RESULTS: Images of PET/CT scans of the shoulder, hip and knee joints revealed that FDG accumulated at the entheses in SpA and in the synovium in RA patients. The maximum SUVs [mean (s.d.)] were statistically higher in SpA patients compared with RA patients at the entheses of lumbar spinous process [4.83 (1.15) vs 1.42 (0.34); P < 0.05, respectively], pubic symphysis [3.93 (0.87) vs 1.35 (0.31); P < 0.05, respectively] and ischial tuberosity [4.76 (1.5) vs 1.35 (0.42); P < 0.05, respectively]. The positive frequencies of lumbar spinous processes and ischial tuberosity evaluated by PET/CT scan in the SpA group were significantly higher than that evaluated by MRI. CONCLUSION: MRI is now widely used to detect bone marrow oedema and enthesitis in patients with SpA. PET/CT scans offer an alternative method to identify enthesitis, and will likely contribute to the early diagnosis of SpA. PMID- 20007286 TI - Age-related T-cell cytokine profile parallels corneal disease severity in Sjogren's syndrome-like keratoconjunctivitis sicca in CD25KO mice. AB - OBJECTIVES: IL-2ralpha (CD25)(-/-) mice develop autoimmunity and lymphoproliferative disorders, including SS-like disease. The objective of this study was to evaluate the severity of corneal epithelial disease and T-cell cytokine profile in the ocular surface tissues of CD25KO mice. METHODS: CD25KO mice were evaluated at 8, 12 and 16 weeks of age. Corneal epithelial smoothness and corneal permeability were measured. Phenotype of infiltrating lymphocytes was evaluated by immunohistochemistry. Th-1, -2 and -17 associated factors were measured by real-time PCR in cornea and conjunctiva and by Luminex immunobead assay in tears. RESULTS: Compared with 8-week-old wild-type (WT) mice, CD25KO mice of the same age had significantly greater corneal irregularity and a significant increase in the number of CD4(+) and CD8(+) T cells infiltrating the conjunctiva. CD25KO mice had significantly higher levels of IL-6, TGF-beta1, IL 23R, IL-17A, IL-17F, IL-21, CCL20, IL-10, GATA-3 and IFN-gamma mRNA transcripts in their cornea and conjunctiva than WT mice at 8 weeks. IL-17A and IL-17F mRNA transcripts peaked at 12 weeks, whereas IFN-gamma spiked at 16 weeks in CD25KO mice. Increased expression of IL-17A and IL-17F at 12 weeks in CD25KO mice was accompanied by a worsening of corneal surface parameters and an increase of CD4(+) T cell infiltrating the cornea. CONCLUSIONS: Disruption of IL-2 signalling in CD25KO mice results in age-dependent SS-like autoimmune lacrimal keratoconjunctivitis. A mix of Th-1 and Th-17 cytokines was detected. The peak severity of corneal epithelial disease corresponded to the peak of IL-17 expression. PMID- 20007288 TI - Heart rate turbulence assessment in systemic sclerosis: the role for the detection of cardiac autonomic nervous system dysfunction. AB - OBJECTIVE: To assess the heart rate turbulence (HRT) in patients with SSc as a method of assessment for cardiac autonomic nervous function. METHODS: We prospectively studied 68 consecutive patients with SSc before inclusion in the study. After a detailed clinical evaluation, including echocardiography, 45 subjects [aged 54.6 (14.7) years; 40 women] underwent 24-h Holter monitoring for HRT and time- and frequency-domain heart rate variability (HRV) assessment. Results were compared with those in 30 age- and sex-matched healthy controls. RESULTS: As compared with controls, HRT was impaired in SSc patients: the median turbulence onset (TO) was higher (P = 0.0001) and the median turbulence slope (TS) was lower (P = 0.0003). Abnormal HRT (TO > or =0.0% and/or TS < or =2.5 ms/RR) was found in 42% of SSc patients. Moreover, SSc duration correlated negatively with values of TS (r = -0.3; P = 0.045). HRT did not differ between diffuse and limited SSc groups. All estimated time- and the majority of frequency domain values of HRV parameters in SSc were significantly lower than in controls. Significant correlations were also demonstrated between HRT and HRV parameters. CONCLUSIONS: HRT, like HRV assessment, indicates a frequent impairment of the cardiac autonomic nervous system in SSc patients, irrespective of the SSc type. PMID- 20007289 TI - Stomatal density is controlled by a mesophyll-derived signaling molecule. AB - Stomata are composed of a pair of guard cells and a pore between them, and their density and positions are regulated by developmental and environmental signals. In a screen in which we overexpressed many genes coding for putative secretory proteins one by one in Arabidopsis, we identified a gene named STOMAGEN, which increases stomatal density when overexpressed. The STOMAGEN gene encodes a small peptide with a putative secretory signal sequence at its N-terminus and is expressed preferentially in mesophyll cells. This peptide belongs to the EPIDERMAL PATTERNING FACTOR (EPF) family of the cysteine-rich peptides superfamily. The mature form was a 45-amino-acid peptide (stomagen) with three intramolecular disulfide bonds. Stomagen treatment at very low concentrations, as low as 10 nM, increased the stomatal density of wild-type Arabidopsis plants. We propose that stomagen is a mesophyll-to-epidermis signaling molecule that positively regulates stomatal density. We also suggest that stomagen increases stomatal density by competing with negative regulators EPF1 and EPF2 for the receptor-like protein TOO MANY MOUTHS. PMID- 20007290 TI - Arabidopsis chloroplastic ascorbate peroxidase isoenzymes play a dual role in photoprotection and gene regulation under photooxidative stress. AB - Though two types of chloroplastic ascorbate peroxidase (APX) located in the thylakoid membrane (tAPX) and stroma (sAPX) have been thought to be key regulators of intracellular levels of H(2)O(2), their physiological significance in the response to photooxidative stress is still under discussion. Here we characterized single mutants lacking either tAPX (KO-tAPX) or sAPX (KO-sAPX). Under exposure to high light or treatment with methylviologen under light, H(2)O(2) and oxidized proteins accumulated to higher levels in both mutant plants than in the wild-type plants. On the other hand, the absence of sAPX and tAPX drastically suppressed the expression of H(2)O(2)-responsive genes under photooxidative stress. Interestingly, the most marked effect of photooxidative stress on the accumulation of H(2)O(2) and oxidized protein and gene expression was observed in the KO-tAPX plants rather than the KO-sAPX plants. The present findings suggest that both chloroplastic APXs, but particularly tAPX, are important for photoprotection and gene regulation under photooxidative stress in Arabidopsis leaves. PMID- 20007292 TI - Plain packaging: a time for action. PMID- 20007291 TI - Inactivation of duplicated nod factor receptor 5 (NFR5) genes in recessive loss of-function non-nodulation mutants of allotetraploid soybean (Glycine max L. Merr.). AB - Chemically induced non-nodulating nod139 and nn5 mutants of soybean (Glycine max) show no visible symptoms in response to rhizobial inoculation. Both exhibit recessive Mendelian inheritance suggesting loss of function. By allele determination and genetic complementation in nod139 and nn5, two highly related lipo-oligochitin LysM-type receptor kinase genes in Glycine max were cloned; they are presumed to be the critical nodulation-inducing (Nod) factor receptor similar to those of Lotus japonicus, pea and Medicago truncatula. These duplicated receptor genes were called GmNFR5alpha and GmNFR5beta. Nonsense mutations in GmNFR5alpha and GmNFR5beta were genetically complemented by both wild-type GmNFR5alpha and GmNFR5beta in transgenic roots, indicating that both genes are functional. Both genes lack introns. In cultivar Williams82 GmNFR5alpha is located in chromosome 11 and in tandem with GmLYK7 (a related LysM receptor kinase gene), while GmNFR5beta is in tandem with GmLYK4 in homologous chromosome 1, suggesting ancient synteny and regional segmental duplication. Both genes are wild type in G. soja CPI100070 and Harosoy63; however, a non-functional NFR5beta allele (NFR5beta*) was discovered in parental lines Bragg and Williams, which harbored an identical 1,407 bp retroelement-type insertion. This retroelement (GmRE-1) and related sequences are located in several soybean genome positions. Paradoxically, putatively unrelated soybean cultivars shared the same insertion, suggesting a smaller than anticipated genetic base in this crop. GmNFR5alpha but not GmNFR5beta* was expressed in inoculated and uninoculated tap and lateral root portions at about 10-25% of GmATS1 (ATP synthase subunit 1), but not in trifoliate leaves and shoot tips. PMID- 20007293 TI - Expressions of cytochrome P450, UDP-glucuronosyltranferase, and transporter genes in monolayer carcinoma cells change in subcutaneous tumors grown as xenografts in immunodeficient nude mice. AB - Human tumors grown as xenografts in immunodeficient nude mice are widely used to investigate the pharmacological activities of anticancer drugs. Drug-metabolizing enzymes and transporters are expressed in tumor cell lines and changes in drug metabolism and pharmacokinetics (DMPK)-related gene expression after inoculation of the tumor cell may affect the pharmacological activity of the drug under consideration. The aims of the current study were to characterize DMPK-related gene expression profiles and responses to typical cytochrome P450 inducers in monolayer carcinoma cells grown in tissue culture versus those inoculated into a xenograft model. We used the human hepatocellular carcinoma cell line PLC/PRF/5 for this study and comprehensively assessed changes in DMPK-related gene expression by reverse transcription-polymerase chain reaction quantitation. CYP3A4 and UDP-glucuronosyltransferase 1A protein amounts were also analyzed by immunoprecipitation followed by immunoblotting. We found that the expression of many DMPK-related genes was elevated in the inoculated tumor compared with the monolayer carcinoma cells, indicating changes in their gene regulation pathways, presumably due to modulation of the nuclear receptor family of transcription factors. In addition, monolayer carcinoma versus inoculated tumor cells showed different responses to rifampicin, but similar responses to dexamethasone or 3 methylcholanthrene. These results suggest that inoculation of tumor cells results in the activation of drug metabolism and transport function, leading to changes in the responses to pregnane X receptor ligands and consequent discrepancies in the pharmacological activities between in vitro monolayer carcinoma cells and in vivo xenograft models. PMID- 20007294 TI - Current cytochrome P450 phenotyping methods applied to metabolic drug-drug interaction prediction in dogs. AB - Recombinant cytochrome P450 (P450) phenotyping, different approaches for estimating fraction metabolized (f(m)), and multiple measures of in vivo inhibitor exposure were tested for their ability to predict drug interaction magnitude in dogs. In previous reports, midazolam-ketoconazole interaction studies in dogs have been attributed to inhibition of CYP3A pathways. However, in vitro phenotyping studies demonstrated higher apparent intrinsic clearances (CL(int,app)) of midazolam with canine CYP2B11 and CYP2C21. Application of activity correction factors and isoform hepatic abundance to liver microsome CL(int,app) values further implicated CYP2B11 (f(m) >or= 0.89) as the dog enzyme responsible for midazolam- and temazepam-ketoconazole interactions in vivo. Mean area under the curve (AUC) in the presence of the inhibitor/AUC ratios from intravenous and oral midazolam interaction studies were predicted well with unbound K(i) and estimates of unbound hepatic inlet inhibitor concentrations and intestinal metabolism using the AUC-competitive inhibitor relationship. No interactions were observed in vivo with bufuralol, although significant interactions with bufuralol were predicted with fluoxetine via CYP2D and CYP2C pathways (>2.45-fold) but not with clomipramine (<2-fold). The minor caffeine fluvoxamine interaction (1.78-fold) was slightly higher than predicted values based on determination of a moderate f(m) value for CYP1A1, although CYP1A2 may also be involved in caffeine metabolism. The findings suggest promise for in vitro approaches to drug interaction assessment in dogs, but they also highlight the need to identify improved substrate and inhibitor probes for canine P450s. PMID- 20007295 TI - Glucuronidation of dihydrotestosterone and trans-androsterone by recombinant UDP glucuronosyltransferase (UGT) 1A4: evidence for multiple UGT1A4 aglycone binding sites. AB - UDP-glucuronosyltransferase (UGT) 1A4-catalyzed glucuronidation is an important drug elimination pathway. Although atypical kinetic profiles (nonhyperbolic, non Michaelis-Menten) of UGT1A4-catalyzed glucuronidation have been reported occasionally, systematic kinetic studies to explore the existence of multiple aglycone binding sites in UGT1A4 have not been conducted. To this end, two positional isomers, dihydrotestosterone (DHT) and trans-androsterone (t-AND), were used as probe substrates, and their glucuronidation kinetics with HEK293 expressed UGT1A4 were evaluated both alone and in the presence of a UGT1A4 substrate [tamoxifen (TAM) or lamotrigine (LTG)]. Coincubation with TAM, a high affinity UGT1A4 substrate, resulted in a concentration-dependent activation/inhibition effect on DHT and t-AND glucuronidation, whereas LTG, a low affinity UGT1A4 substrate, noncompetitively inhibited both processes. The glucuronidation kinetics of TAM were then evaluated both alone and in the presence of different concentrations of DHT or t-AND. TAM displayed substrate inhibition kinetics, suggesting that TAM may have two binding sites in UGT1A4. However, the substrate inhibition kinetic profile of TAM became more hyperbolic as the DHT or t-AND concentration was increased. Various two-site kinetic models adequately explained the interactions between TAM and DHT or TAM and t-AND. In addition, the effect of TAM on LTG glucuronidation was evaluated. In contrast to the mixed effect of TAM on DHT and t-AND glucuronidation, TAM inhibited LTG glucuronidation. Our results suggest that multiple aglycone binding sites exist within UGT1A4, which may result in atypical kinetics (both homotropic and heterotropic) in a substrate-dependent fashion. PMID- 20007296 TI - Down-regulation of liver drug-metabolizing enzymes in a murine model of chronic renal failure. AB - Drug metabolism could be altered in patients with chronic renal failure (CRF). In rats, this phenomenon is related to a decrease in liver cytochrome P450 (P450) and phase II enzymes, particularly N-acetyltransferase 2 (NAT2). This study attempted to determine the effects of CRF on liver P450 isoforms and NAT2 expressions by using a CRF mouse model. Two groups of mice were studied: CRF induced by 3/4 nephrectomy and control. Liver protein expression and mRNA levels of the major P450 isoforms involved in drug metabolism (CYP1A2, 2C29, 2D, 2E1, and 3A11) and NAT2 were measured by Western blot and real-time polymerase chain reaction (PCR), respectively. CYP3A activity was also assessed by the N demethylation of erythromycin. Results showed a significant reduction in the protein expression of CYP1A2 (56%), 2C29 (31%), and 3A11 (37%) in CRF mice compared with control animals. Real-time PCR revealed a similar reduction in mRNA levels of CYP1A2, 2C29, and 3A11 (59, 56, and 37%, respectively), in CRF mice. There was no significant modification in protein expression and mRNA of CYP2D and 2E1. Compared with control animals, CRF mice displayed a 25% reduction in N demethylation of erythromycin. For NAT2, protein expression decreased by 33% and mRNA levels decreased by 23%. In conclusion, this study demonstrates that protein expression of liver CYP1A2, CYP2C29, and CYP3A11 is down-regulated in CRF mice, secondary to reduced gene expression. Phase II enzymes are similarly affected by CRF. Our results will allow the use of knockout mice to determine the mechanism underlying CRF-induced down-regulation of liver drug-metabolizing enzymes. PMID- 20007297 TI - UDP-glucuronosyltransferase 1A10: activity against the tobacco-specific nitrosamine, 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanol, and a potential role for a novel UGT1A10 promoter deletion polymorphism in cancer susceptibility. AB - The extrahepatic UDP-glucuronosyltransferase 1A10 (UGT1A10) is a phase II metabolizing enzyme that is active against a number of potent carcinogens. In the present study, UGT1A10 was examined for activity against 4-(methylnitrosamino)-1 (3-pyridyl)-1-butanol (NNAL), the major procarcinogenic metabolite of the potent tobacco-specific nitrosamine 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone, and the promoter region of UGT1A10 was examined for variants that could lead to altered UGT1A10 expression. UGT1A10-overexpressing cell homogenates exhibited high O-glucuronidation activity against NNAL (K(M) = 5.95 mM). A 2000-base pair (bp) product corresponding to the UGT1A10 proximal promoter region was polymerase chain reaction (PCR)-amplified using genomic DNA from 97 white subjects, and 42 of these were sequenced. In addition to a previously reported C/G single nucleotide polymorphism at -1271 bp (rs2741032), a novel 1664-bp deletion located between nucleotides -190 to -1856 relative to the UGT1A10 translation start site was identified. Using real-time multiplex PCR, this deletion exhibited a prevalence of 0.022 in whites (n = 156) and 0.056 in blacks (n = 133). To determine whether either polymorphism altered gene expression, in vitro assays were performed using luciferase constructs containing up to 2000 bp of the proximal UGT1A10 promoter. Constructs containing the 1664-bp deletion exhibited a significant (p = 0.009) 3-fold increase in luciferase activity compared with constructs containing the wild-type UGT1A10 promoter. No effect on luciferase activity was observed for the UGT1A10(-1271G) promoter variant. These data are consistent with previous studies that indicate the presence of a transcriptional repressor element within the newly identified deletion and that this deletion polymorphism may contribute to altered UGT1A10 expression and altered carcinogen detoxification between individuals. PMID- 20007299 TI - Dynamics and heterogeneity in the process of human frailty and aging: evidence from the U.S. older adult population. AB - OBJECTIVES: This study investigated the dynamics and heterogeneity of the frailty index (FI) conceived as a systemic indicator of biological aging in the community dwelling older adult population in the United States. METHODS: We used panel data on multiple birth cohorts from the Health and Retirement Survey 1993-2006 and growth curve models to estimate age trajectories of the FI and their differences by sex, race, and socioeconomic status (SES) within cohorts. RESULTS: The FI for cohorts born before 1942 exhibit quadratic increases with age and accelerated increases in the accumulation of health deficits. More recent cohorts exhibit higher average levels of and rates of increment in the FI than their predecessors do at the same ages. Females, non-Whites, and individuals with low education and income exhibit greater degrees of physiological deregulation than their male, White, and high-SES counterparts at any age. Patterns of sex, race, and SES differentials in rates of aging vary across cohorts. DISCUSSION: Adjusting for social behavioral factors, the analysis provides evidence for physiological differences in the aging process among recent cohorts of older adults, points to the need for biological explanations of female excess in general system damage, and reveals the insufficiency of any single mechanism for depicting the racial and SES differences in the process of physiological deterioration. PMID- 20007300 TI - It is not getting old: the rejuvenation of social gerontology. PMID- 20007298 TI - Transcription coactivator PBP/MED1-deficient hepatocytes are not susceptible to diethylnitrosamine-induced hepatocarcinogenesis in the mouse. AB - Nuclear receptor coactivator [peroxisome proliferator-activated receptor-binding protein (PBP)/mediator subunit 1 (MED1)] is a critical component of the mediator transcription complex. Disruption of this gene in the mouse results in embryonic lethality. Using the PBP/MED1 liver conditional null (PBP/MED1(DeltaLiv)) mice, we reported that PBP/MED1 is essential for liver regeneration and the peroxisome proliferator-activated receptor alpha ligand Wy-14,643-induced receptor-mediated hepatocarcinogenesis. We now examined the role of PBP/MED1 in genotoxic chemical carcinogen diethylnitrosamine (DEN)-induced and phenobarbital-promoted hepatocarcinogenesis. The carcinogenic process was initiated by a single intraperitoneal injection of DEN at 14 days of age and initiated cells were promoted with phenobarbital (PB) (0.05%) in drinking water. PBP/MED1(DeltaLiv) mice, killed at 1, 4 and 12 weeks, revealed a striking proliferative response of few residual PBP/MED1-positive hepatocytes that escaped Cre-mediated deletion of PBP/MED1 gene. No proliferative expansion of PBP/MED1 null hepatocytes was noted in the PBP/MED1(DeltaLiv) mouse livers. Multiple hepatocellular carcinomas (HCCs) developed in the DEN-initiated PBP/MED1(fl/fl) and PBP/MED1(DeltaLiv) mice, 1 year after the PB promotion. Of interest is that all HCC developing in PBP/MED1(DeltaLiv) mice were PBP/MED1 positive. None of the tumors was PBP/MED1 negative implying that hepatocytes deficient in PBP/MED1 are not susceptible to neoplastic conversion. HCC that developed in PBP/MED1(DeltaLiv) mouse livers were transplantable in athymic nude mice and these maintained PBP/MED1(fl/fl) genotype. PBP/MED1(fl/fl) HCC cell line derived from these tumors expressed PBP/MED1 and deletion of PBP/MED1(fl/fl) allele by adeno-Cre injection into tumors caused necrosis of tumor cells. These results indicate that PBP/MED1 is essential for the development of HCC in the mouse. PMID- 20007301 TI - Religion and mental health among older adults: do the effects of religious involvement vary by gender? AB - OBJECTIVES: Few studies explore how the relationship between religious involvement and mental health varies by gender among the aging population. This article outlines a series of arguments concerning the effects of gender in moderating the effect of religious involvement on mental health and examines them empirically. METHODS: Using two waves (2001 and 2004) of the Religion, Aging, and Health Survey, this study estimates the differential effect of gender in the religion-mental health connection using multivariate analyses for a nationally representative sample of U.S. adults aged 66-95 years. RESULTS: Results suggest that (a) men obtain more mental health benefits from religious involvement than women, (b) women with higher levels of organizational religious involvement have similar levels of mental health as those with moderate and lower levels of organizational religious involvement, (c) men with very high levels of organizational religious involvement tend to have much higher levels of mental health than all other men. DISCUSSION: The relationship between organizational religious involvement and mental health is found to be mostly a nonlinear one such that those with the highest levels of religiosity receive all the benefits. The findings suggest a number of promising research directions on the religion mental health connection among older Americans. PMID- 20007302 TI - Folate intake, post-folic acid grain fortification, and pancreatic cancer risk in the Prostate, Lung, Colorectal, and Ovarian Cancer Screening Trial. AB - BACKGROUND: Folate plays a critical role in DNA methylation, synthesis, and repair. Several epidemiologic studies suggest that higher folate intake is associated with decreased pancreatic cancer risk. OBJECTIVE: We investigated the association between dietary folate intake and pancreatic cancer in the Prostate, Lung, Colorectal, and Ovarian Cancer Screening Trial (PLCO) cohort. DESIGN: Dietary data were collected with the use of a self-administered food-frequency questionnaire (1998-2005). Among the 51,988 male and 57,187 female participants, aged 55-74 y at enrollment, with complete dietary and multivitamin information, 162 men and 104 women developed pancreatic cancer during follow-up (January 1998 to December 2006; median: 6.5 y). We used Cox proportional hazards regression with age as the time metric to calculate hazard ratios (HRs) and 95% CIs. RESULTS: The highest compared with the lowest quartile of food folate was associated with a significantly decreased pancreatic cancer risk among women (> or = 253.3 compared with < or = 179.1 microg/d; HR = 0.47; 95% CI: 0.23, 0.94; P for trend: 0.09) but not among men (> or = 229.6 compared with < or = 158.0 microg/d; HR = 1.20; 95% CI: 0.70, 2.04; P for trend: 0.67; P for interaction by sex: 0.03). There was also a significant inverse trend in risk of pancreatic cancer across increasing quartiles of total folate in women (P for trend: 0.04) but not in men (P for trend: 0.65). Folic acid supplements were not associated with pancreatic cancer. CONCLUSION: These findings support an association between higher food and total folate intakes and decreased risk of pancreatic cancer in women but not in men. PMID- 20007303 TI - Breast, colorectal, and prostate cancer risk in the European Prospective Investigation into Cancer and Nutrition-Norfolk in relation to phytoestrogen intake derived from an improved database. AB - BACKGROUND: The characterization of phytoestrogen intake and cancer risk has been hindered by the absence of accurate dietary phytoestrogen values. OBJECTIVE: We examined the risk of breast, colorectal, and prostate cancers relative to phytoestrogen intake on the basis of a comprehensive database. DESIGN: Demographic and anthropometric characteristics, a medical history, and 7-d records of diet were collected prospectively from participants (aged 40-79 y) in the European Prospective Investigation into Cancer and Nutrition-Norfolk (EPIC Norfolk). Five hundred nine food items were analyzed by liquid chromatography mass spectrometry/mass spectrometry, and (13)C(3)-labeled internal standards were analyzed for isoflavones (genistein, daidzein, glycitein, biochanin A, and formononetin), lignans (secoisolariciresinol and matairesinol), and enterolignans from gut microbial metabolism in animal food sources (equol and enterolactone). From the direct analysis, values for 10,708 foods were calculated. Odds ratios (ORs) for breast (244 cases, 941 controls), colorectal (221 cases, 886 controls), and prostate (204 cases, 812 controls) cancers were calculated relative to phytoestrogen intake. RESULTS: Phytoestrogen intake was not associated with breast cancer among women or colorectal cancer among men. Among women, colorectal cancer risk was inversely associated with enterolactone (OR: 0.33; 95% CI: 0.14, 0.74) and total enterolignans (OR: 0.32; 95% CI: 0.13, 0.79), with a positive trend detected for secoisolariciresinol (OR: 1.60; 95% CI: 0.96, 2.69). A positive trend between enterolignan intake and prostate cancer risk (OR: 1.27; 95% CI: 0.97, 1.66) was attenuated after adjustment for dairy intake (OR: 1.19; 95% CI: 0.77, 1.82). CONCLUSION: Dietary phytoestrogens may contribute to the risk of colorectal cancer among women and prostate cancer among men. PMID- 20007304 TI - Adherence to a Mediterranean diet and risk of gastric adenocarcinoma within the European Prospective Investigation into Cancer and Nutrition (EPIC) cohort study. AB - BACKGROUND: The Mediterranean dietary pattern is believed to protect against cancer, although evidence from cohort studies that have examined particular cancer sites is limited. OBJECTIVE: We aimed to explore the association between adherence to a relative Mediterranean diet (rMED) and incident gastric adenocarcinoma (GC) within the European Prospective Investigation into Cancer and Nutrition study. DESIGN: The study included 485,044 subjects (144,577 men) aged 35-70 y from 10 European countries. At recruitment, dietary and lifestyle information was collected. An 18-unit rMED score, incorporating 9 key components of the Mediterranean diet, was used to estimate rMED adherence. The association between rMED and GC with respect to anatomic location (cardia and noncardia) and histologic types (diffuse and intestinal) was investigated. A calibration study in a subsample was used to control for dietary measurement error. RESULTS: After a mean follow-up of 8.9 y, 449 validated incident GC cases were identified and used in the analysis. After stratification by center and age and adjustment for recognized cancer risk factors, high compared with low rMED adherence was associated with a significant reduction in GC risk (hazard ratio: 0.67; 95% CI: 0.47, 0.94). A 1-unit increase in the rMED score was associated with a decreased risk of GC of 5% (95% CI: 0.91, 0.99). There was no evidence of heterogeneity between different anatomic locations or histologic types. The calibrated results showed similar trends (overall hazard ratio for GC: 0.93; 95% CI: 0.89, 0.99). CONCLUSION: Greater adherence to an rMED is associated with a significant reduction in the risk of incident GC. PMID- 20007305 TI - Offering choice and its effect on Dutch children's liking and consumption of vegetables: a randomized controlled trial. AB - BACKGROUND: Children's vegetable consumption is below recommended amounts. According to self-determination theory, stimulating children's feelings of autonomy by offering a choice of vegetables may be a valuable strategy to increase their vegetable liking and consumption. The effect of choice-offering on children's vegetable liking and consumption has, to our knowledge, not yet been studied. OBJECTIVE: The objective was to investigate whether having a choice between 2 vegetables enhances children's vegetable liking and consumption. DESIGN: Three hundred three children (age: 4-6 y) were randomly assigned to receive 1 of 3 dinner conditions in a restaurant setting. Two similarly liked vegetables were presented, after which the child had no choice, a premeal choice, or an at-meal choice. Subsequently, the dinner was consumed with one parent present. Comparisons between the 3 conditions regarding children's meal experience, vegetable liking, and vegetable consumption were made by using analysis of variance and Kruskal-Wallis tests. RESULTS: Children's vegetable consumption did not differ (P = 0.54) between the conditions as follows: 56 + or 45 g in the no-choice condition, 51 + or - 46 g in the premeal-choice condition, and 49 + or - 47 g in the at-meal-choice condition. In the no-choice condition, high-reactant children (who are more sensitive to psychological, persuasive pressure) consumed fewer vegetables (45 + or - 42 g) than did low-reactant children (73 + or - 43 g; P = 0.04). Vegetable liking was similar in all 3 conditions (P = 0.43). Children appreciated being able to choose in the premeal choice condition. CONCLUSIONS: A premeal choice between 2 vegetables was appreciated by the children but did not increase their vegetable liking and consumption. The no-choice condition decreased vegetable consumption in high reactant children. Future research should investigate the effects of choice offering in the long term and in more familiar eating settings. This trial was registered at controlled-trials.com as ISRCTN03035138. PMID- 20007306 TI - Ghrelin and peptide YY in postpartum lactating and nonlactating women. AB - BACKGROUND: Epidemiologic studies suggest that childbearing is an important contributor to the development of obesity in many women and that breastfeeding may be protective. Ghrelin and peptide YY (PYY) are gut hormones involved in appetite regulation and energy homeostasis and are biological neuroendocrine signals that potentially affect body weight and adiposity. OBJECTIVE: This study evaluated whether fasting or postprandial ghrelin or PYY is different between lactating and nonlactating postpartum women matched for age, body weight, and adiposity. DESIGN: Ten postpartum lactating women (mean + or - SD: 28.1 + or - 4.9 y of age, 69.2 + or - 11.3 kg, 35.4 + or - 6.6% body fat) and 8 nonlactating women (28.8 + or - 7.6 y of age, 75.6 + or - 13.7 kg, 37.5 + or - 6.5% body fat) at 4-5 wk postpartum underwent measurements of body weight, body composition, and ghrelin and PYY responses to a standardized meal (350 kcal). Seven never-pregnant women served as control subjects (29.7 + or - 4.1 y of age, 60.4 + or - 4.8 kg, 25.5 + or - 2.0% body fat). RESULTS: Ghrelin concentrations decreased, whereas PYY concentrations increased significantly (P < 0.05) in response to the meal, but fasting or meal-induced changes were not significantly different between lactating and nonlactating women. The fasting ghrelin concentration correlated with body mass index (r = -0.53, P < 0.05) and was significantly lower in postpartum than in control women (894.9 + or - 247.7 compared with 1316.9 + or - 241.0 pg/mL), even after adjustment for body mass index. CONCLUSIONS: Our data do not support the notion that ghrelin, PYY, or both are plausible neuroendocrine signals that influence body weight regulation during lactation. They suggest, however, that ghrelin may change with increased adiposity in the postpartum state and may potentially play a role in body weight regulation after child birth. PMID- 20007307 TI - Racial differences in potassium homeostasis in response to differences in dietary sodium in girls. AB - BACKGROUND: Racial differences in the renal disposition of potassium may be related to mechanisms for the greater susceptibility to hypertension in blacks than in whites. OBJECTIVE: Our objective was to study the racial differences in the renin-angiotensin-aldosterone system and in potassium balance in black and white girls consuming a controlled diet that was low in potassium with 2 amounts of sodium intake (low compared with high). DESIGN: The studies reported here were performed in 40 black and 28 white girls, aged 11-15 y, under highly controlled metabolic conditions. The studies comprised 2 sessions of 20-d metabolic balance sessions, at 2 amounts of dietary sodium intake (58 and 170 mmol . L(-1) . d( 1)), in a crossover design and with a constant dietary potassium intake of 50 mmol . L(-1) . d(-1). Repeated-measures analysis of variance was used to test for racial differences in potassium output and retention by sodium intakes. RESULTS: Thirty black and 20 white girls completed the study. Urinary potassium excretion was lower in blacks than in whites, regardless of sodium intake (P < 0.05), with no differences in fecal or sweat potassium excretion. Cumulative potassium retention was significantly higher in blacks while consuming the low sodium diet. Plasma aldosterone concentrations after upright posture were significantly lower in blacks than in whites but were similar when supine, as were urinary aldosterone excretion rates. On week 3, blood pressure, body weight, urinary volume, creatinine, and serum sodium and potassium were similar. CONCLUSION: The well-known racial difference in urinary potassium excretion appears to be at least in part due to greater renal retention of potassium in black girls. PMID- 20007308 TI - Influence of common variants near INSIG2, in FTO, and near MC4R genes on overweight and the metabolic profile in adolescence: the TRAILS (TRacking Adolescents' Individual Lives Survey) Study. AB - BACKGROUND: Overweight is a complex trait in which both environmental and genetic factors play a role. OBJECTIVE: We aimed to evaluate the influence of common genetic variants identified by genome-wide association studies on overweight and the metabolic profile in adolescence. DESIGN: In a population-based cohort of 663 girls and 612 boys aged 16 y, weight, height, skinfold thicknesses, percentage body fat, waist circumference, blood pressure, glucose, insulin, lipid profile, and DNA were obtained. We defined overweight according to international criteria. We performed multiple linear and logistic regression analyses to assess the influence of candidate single nucleotide polymorphisms near the INSIG2, in the FTO, and near the MC4R genes and repeated-measures analyses of available body mass index (BMI) and skinfold thickness data across 3 visits at ages 11, 13.5, and 16 y. RESULTS: A total of 15.1% of participants were overweight or obese at age 16 y. No associations with INSIG2 were found. Common variation in the FTO gene was associated with sex-specific z scores of BMI (B: 0.11; 95% CI: 0.03, 0.19), sum of skinfold thicknesses (B: 0.12; 95% CI: 0.04, 0.20), percentage body fat (B: 0.11; 95% CI: 0.03, 0.19), waist circumference (B: 0.11; 95% CI: 0.03, 0.19), fasting glucose (B: 0.10; 95% CI: 0.00, 0.20), and overweight (odds ratio: 1.34; 95% CI: 1.06, 1.69) at age 16 y. Repeated-measures analyses confirmed the associations for BMI and sum of skinfold thicknesses, and physical activity did not modify these associations. Common variation near the MC4R gene was associated with BMI in cross-sectional (B: 0.11; 95% CI: 0.02, 0.20) and repeated-measures (B: 0.12; 95% CI: 0.03, 0.20) analyses. CONCLUSIONS: Common variation in the FTO gene is associated with overall and abdominal adiposity. Variation near the MC4R gene is associated with BMI. These findings in adolescents strengthen and extend the results from previous research. PMID- 20007309 TI - Chemokines in the vascular inflammatory response of atherosclerosis. AB - Atherosclerosis is considered to be a chronic inflammatory disease of the vessel wall that encompasses the accumulation of lipids, and it is critically shaped by the recruitment of leucocytes during all phases of the disease. In addition, the progression of atherosclerosis is determined by a disturbed equilibrium of immune responses. Chemokines and their receptors are instrumental in orchestrating the influx of leucocytes to the vascular wall, but also seem to regulate immune functions. Recent work has shed light on the apparent redundancy and the robustness of the chemokine system and has also provided evidence for its specialized role in the regulation of specific functions and trafficking of leucocyte subpopulations. This review will give a comprehensive summary to highlight those chemokines addressed in different models of atherosclerosis and vascular injury to date. In addition, we will discuss recent developments scrutinizing heterophilic interactions of chemokines that have advanced our understanding of how chemokines control vascular inflammatory responses. PMID- 20007310 TI - Early exercise training after myocardial infarction prevents contractile but not electrical remodelling or hypertrophy. AB - AIMS: Exercise started early after myocardial infarction (MI) improves in vivo cardiac function and myofilament responsiveness to Ca(2+). We investigated whether this represents partial or complete reversal of cellular remodelling. METHODS AND RESULTS: Mice with MI following left coronary ligation were given free access to a running wheel (MI(EXE), N = 22) or housed sedentary (MI(SED), N = 18) for 8 weeks and compared with sedentary sham-operated animals (SHAM, N = 11). Myocytes were enzymatically isolated from the non-infarcted left ventricle. Myocytes in MI were significantly longer and even more so with exercise (165 +/- 3 microm in MI(EXE) vs. 148 +/- 3 microm in MI(SED) and 136 +/- 2 microm in SHAM; P < 0.05, mean +/- SEM); cell width was not different. Contraction was measured during electrical field stimulation at 1, 2, and 4 Hz. Unloaded cell shortening was significantly reduced in MI(SED) (at 1 Hz, L/L(0)=4.4 +/- 0.3% vs. 6.7 +/- 0.4% in SHAM; P < 0.05, also at 2 and 4 Hz). Exercise restored cell shortening to SHAM values (MI(EXE), L/L(0)=6.4 +/- 0.5%). Membrane currents and [Ca(2+)](i) were measured via whole-cell patch clamping, with Fluo-3 as Ca(2+) indicator, all at 30 degrees C. Ca(2+) transient amplitude, I(CaL) and sarcoplasmic reticulum Ca(2+) content were not different between the three groups. Diastolic Ca(2+) levels at 4 Hz were significantly elevated in MI(SED) only, with a trend to increased spontaneous Ca(2+) release events (sparks). Action potential duration was increased and transient outward K(+) currents significantly reduced after MI; this was unaffected by exercise. CONCLUSIONS: Early voluntary exercise training after MI restores cell contraction to normal values predominantly because of changes in the myofilament Ca(2+) response and has a beneficial effect on diastolic Ca(2+) handling. However, the beneficial effect is not a complete reversal of remodelling as hypertrophy and loss of repolarizing K(+) currents are not affected. PMID- 20007311 TI - Desensitization of vascular endothelin receptors by G protein-coupled receptor kinase 2. PMID- 20007317 TI - Structural basis for the interaction between yeast Spt-Ada-Gcn5 acetyltransferase (SAGA) complex components Sgf11 and Sus1. AB - Sus1 is a central component of the yeast gene gating machinery, the process by which actively transcribing genes such as GAL1 become associated with nuclear pore complexes. Sus1 is a component of both the SAGA transcriptional co-activator complex and the TREX-2 complex that binds to nuclear pore complexes. TREX-2 contains two Sus1 chains that have an articulated helical hairpin fold, enabling them to wrap around an extended alpha-helix in Sac3, following a helical hydrophobic stripe. In SAGA, Sus1 binds to Sgf11 and has been proposed to provide a link between SAGA and TREX-2. We present here the crystal structure of the complex between Sus1 and the N-terminal region of Sgf11 that forms an extended alpha-helix around which Sus1 wraps in a manner that shares some similarities with the Sus1-Sac3 interface in TREX-2. However, the Sus1-binding site on Sgf11 is somewhat shorter than on Sac3 and is based on a narrower hydrophobic stripe. Engineered mutants that disrupt the Sgf11-Sus1 interaction in vitro confirm the importance of the hydrophobic helical stripe in molecular recognition. Helix alpha1 of the Sus1-articulated hairpin does not bind directly to Sgf11 and adopts a wide range of conformations within and between crystal forms, consistent with the presence of a flexible hinge and also with results from previous extensive mutagenesis studies (Klockner, C., Schneider, M., Lutz, S., Jani, D., Kressler, D., Stewart, M., Hurt, E., and Kohler, A. (2009) J. Biol. Chem. 284, 12049 12056). A single Sus1 molecule cannot bind Sgf11 and Sac3 simultaneously and this, combined with the structure of the Sus1-Sgf11 complex, indicates that Sus1 forms separate subcomplexes within SAGA and TREX-2. PMID- 20007318 TI - Zili inhibits transforming growth factor-beta signaling by interacting with Smad4. AB - Piwi proteins are required for germ cell proliferation, differentiation, and germ line stem cell maintenance. In normal tissues, human and mouse Piwil2 are primarily expressed in testis but widely expressed in tumors. However, the underlying mechanism remains largely unknown. In vertebrates, transforming growth factor (TGF)-beta signaling plays an important role in patterning embryo and control of cell growth and differentiation. A previous study has shown a role for Zili, a Piwil2 gene in zebrafish, in germ cells in zebrafish. Here we report that zili functions in patterning the early embryo and inhibits TGF-beta signaling. Whole mount expression analysis shows that zili expresses not only in PGCs but also in axis. Ectopic expression of zili causes fusion of the eyes and reduction of mesodermal marker genes expression, suggesting that zili functions to inhibit Nodal signaling and mesoderm formation. Genetic interaction shows that zili inhibits Nodal and bone morphogenetic protein signaling. The results of protein interaction assays identify that Zili binds to Smad4 via its N-terminal domain and prevents the formation of Smad2/3/4 and Smad1/5/9/4 complexes to antagonize TGF-beta signaling. This work shows that zili plays a role in early embryogenesis beyond germ line as a novel negative regulator of TGF-beta signaling, extending the function of Piwi proteins in vertebrates. PMID- 20007319 TI - A large intrinsically disordered region in SKIP and its disorder-order transition induced by PPIL1 binding revealed by NMR. AB - Intrinsically disordered proteins or protein regions play an important role in fundamental biological processes. During spliceosome activation, a large structural rearrangement occurs. The Prp19 complex and related factors are involved in the catalytic activation of the spliceosome. Recent mass spectrometric analyses have shown that Ski interaction protein (SKIP) and peptidylprolyl isomerase-like protein 1 (PPIL1) are Prp19-related factors that constitute the spliceosome B, B*, and C complexes. Here, we report that a highly flexible region of SKIP (SKIPN, residues 59-129) is intrinsically disordered. Upon binding to PPIL1, SKIPN undergoes a disorder-order transition. A highly conserved fragment of SKIP (residues 59-79) called the PPIL1-binding fragment (PBF) was sufficient to bind PPIL1. The structure of PBF.PPIL1 complex, solved by NMR, shows that PBF exhibits an ordered structure and interacts with PPIL1 through electrostatic and hydrophobic interactions. Three subfragments in the PBF (residues 59-67, 68-73, and 74-79) show hook-like backbone structure, and interactions between these subfragments are necessary for PBF.PPIL1 complex formation. PPIL1 is a cyclophilin family protein. It is recruited by SKIP into the spliceosome by a region other than the peptidylprolyl isomerase active site. This enables the active site of PPIL1 to remain open in the complex and still function as a peptidylprolyl cis/trans-isomerase or molecular chaperon to facilitate the folding of other proteins in the spliceosomes. The large disordered region in SKIP provides an interaction platform. Its disorder-order transition, induced by PPIL1 binding, may adapt the requirement for a large structural rearrangement occurred in the activation of spliceosome. PMID- 20007320 TI - Post-translational modification of ribosomal proteins: structural and functional characterization of RimO from Thermotoga maritima, a radical S-adenosylmethionine methylthiotransferase. AB - Post-translational modifications of ribosomal proteins are important for the accuracy of the decoding machinery. A recent in vivo study has shown that the rimO gene is involved in generation of the 3-methylthio derivative of residue Asp 89 in ribosomal protein S12 (Anton, B. P., Saleh, L., Benner, J. S., Raleigh, E. A., Kasif, S., and Roberts, R. J. (2008) Proc. Natl. Acad. Sci. U. S. A. 105, 1826-1831). This reaction is formally identical to that catalyzed by MiaB on the C2 of adenosine 37 near the anticodon of several tRNAs. We present spectroscopic evidence that Thermotoga maritima RimO, like MiaB, contains two [4Fe-4S] centers, one presumably bound to three invariant cysteines in the central radical S adenosylmethionine (AdoMet) domain and the other to three invariant cysteines in the N-terminal UPF0004 domain. We demonstrate that holo-RimO can specifically methylthiolate the aspartate residue of a 20-mer peptide derived from S12, yielding a mixture of mono- and bismethylthio derivatives. Finally, we present the 2.0 A crystal structure of the central radical AdoMet and the C-terminal TRAM (tRNA methyltransferase 2 and MiaB) domains in apo-RimO. Although the core of the open triose-phosphate isomerase (TIM) barrel of the radical AdoMet domain was conserved, RimO showed differences in domain organization compared with other radical AdoMet enzymes. The unusually acidic TRAM domain, likely to bind the basic S12 protein, is located at the distal edge of the radical AdoMet domain. The basic S12 protein substrate is likely to bind RimO through interactions with both the TRAM domain and the concave surface of the incomplete TIM barrel. These biophysical results provide a foundation for understanding the mechanism of methylthioation by radical AdoMet enzymes in the MiaB/RimO family. PMID- 20007321 TI - Negative autoregulation of GTF2IRD1 in Williams-Beuren syndrome via a novel DNA binding mechanism. AB - The GTF2IRD1 gene is of principal interest to the study of Williams-Beuren syndrome (WBS). This neurodevelopmental disorder results from the hemizygous deletion of a region of chromosome 7q11.23 containing 28 genes including GTF2IRD1. WBS is thought to be caused by haploinsufficiency of certain dosage sensitive genes within the deleted region, and the feature of supravalvular aortic stenosis (SVAS) has been attributed to reduced elastin caused by deletion of ELN. Human genetic mapping data have implicated two related genes GTF2IRD1 and GTF2I in the cause of some the key features of WBS, including craniofacial dysmorphology, hypersociability, and visuospatial deficits. Mice with mutations of the Gtf2ird1 allele show evidence of craniofacial abnormalities and behavioral changes. Here we show the existence of a negative autoregulatory mechanism that controls the level of GTF2IRD1 transcription via direct binding of the GTF2IRD1 protein to a highly conserved region of the GTF2IRD1 promoter containing an array of three binding sites. The affinity for this protein-DNA interaction is critically dependent upon multiple interactions between separate domains of the protein and at least two of the DNA binding sites. This autoregulatory mechanism leads to dosage compensation of GTF2IRD1 transcription in WBS patients. The GTF2IRD1 promoter represents the first established in vivo gene target of the GTF2IRD1 protein, and we use it to model its DNA interaction capabilities. PMID- 20007322 TI - Comparative structural biology of eubacterial and archaeal oligosaccharyltransferases. AB - Oligosaccharyltransferase (OST) catalyzes the transfer of an oligosaccharide from a lipid donor to an asparagine residue in nascent polypeptide chains. In the bacterium Campylobacter jejuni, a single-subunit membrane protein, PglB, catalyzes N-glycosylation. We report the 2.8 A resolution crystal structure of the C-terminal globular domain of PglB and its comparison with the previously determined structure from the archaeon Pyrococcus AglB. The two distantly related oligosaccharyltransferases share unexpected structural similarity beyond that expected from the sequence comparison. The common architecture of the putative catalytic sites revealed a new catalytic motif in PglB. Site-directed mutagenesis analyses confirmed the contribution of this motif to the catalytic function. Bacterial PglB and archaeal AglB constitute a protein family of the catalytic subunit of OST along with STT3 from eukaryotes. A structure-aided multiple sequence alignment of the STT3/PglB/AglB protein family revealed three types of OST catalytic centers. This novel classification will provide a useful framework for understanding the enzymatic properties of the OST enzymes from Eukarya, Archaea, and Bacteria. PMID- 20007323 TI - Ligand-bound structures provide atomic snapshots for the catalytic mechanism of D amino acid deacylase. AB - D-tyrosyl-tRNA(Tyr) deacylase (DTD) is an editing enzyme that removes D-amino acids from mischarged tRNAs. We describe an in-depth analysis of the malaria parasite Plasmodium falciparum DTD here. Our data provide structural insights into DTD complexes with adenosine and D-amino acids. Bound adenosine is proximal to the DTD catalysis site, and it represents the authentic terminal adenosine of charged tRNA. DTD-bound D-amino acids cluster at three different subsites within the overall active site pocket. These subsites, called transition, active, and exit subsites allow docking, re-orientation, chiral selection, catalysis, and exit of the free D-amino acid from DTD. Our studies reveal variable modes of D amino acid recognition by DTDs, suggesting an inherent plasticity that can accommodate all D-amino acids. An in-depth analysis of native, ADP-bound, and D amino acid-complexed DTD structures provide the first atomic snapshots of ligand recognition and subsequent catalysis by this enzyme family. We have mapped sites for the deacylation reaction and mark possible routes for entry and egress of all substrates and products. We have also performed structure-based inhibitor discovery and tested lead compounds against the malaria parasite P. falciparum using growth inhibition assays. Our studies provide a comprehensive structural basis for the catalytic mechanism of DTD enzymes and have implications for inhibition of this enzyme in P. falciparum as a route to inhibiting the parasite. PMID- 20007324 TI - Stem-like cancer cells are inducible by increasing genomic instability in cancer cells. AB - The existence of cancer stem cells (CSCs) or stem-like cancer cells (SLCCs) is regarded as the cause of tumor formation and recurrence. However, the origin of such cells remains controversial with two competing hypotheses: CSCs are either transformed from tissue adult stem cells or dedifferentiated from transformed progenitor cells. Compelling evidence has determined the chromosomal aneuploidy to be one of the hallmarks of cancer cells, indicating genome instability plays an important role in tumorigenesis, for which CSCs are believed to be the initiator. To gain direct evidence that genomic instability is involved in the induction of SLCCs, we utilized multiple approaches to enhance genomic instability and monitored the percentage of SLCC in cultured cancer cells. Using side population (SP) cells as a marker for SLCC in human nasopharyngeal carcinoma (NPC) and CD133 for human neuroblastoma cells, we found that DNA damage inducers, UV and mitomycin C were capable of increasing SP cells in NPC CNE-2 and neuroblastoma SKN-SH cells. Likewise, either overexpression of a key regulator of cell cycle, Mad2, or knock down of Aurora B, an important kinase in mitosis, or Cdh1, a key E3 ligase in cell cycle, resulted in a significant increase of SP cells in CNE-2. More interestingly, enrichment of SP cells was observed in recurrent tumor tissues as compared with the primary tumor in the same NPC patients. Our study thus suggested that, beside transformation of tissue stem cells leading to CSC generation, genomic instability could be another potential mechanism resulting in SLCC formation, especially at tumor recurrence stage. PMID- 20007325 TI - Crystal structure of plant ferritin reveals a novel metal binding site that functions as a transit site for metal transfer in ferritin. AB - Ferritins are important iron storage and detoxification proteins that are widely distributed in living kingdoms. Because plant ferritin possesses both a ferroxidase site and a ferrihydrite nucleation site, it is a suitable model for studying the mechanism of iron storage in ferritin. This article presents for the first time the crystal structure of a plant ferritin from soybean at 1.8-A resolution. The soybean ferritin 4 (SFER4) had a high structural similarity to vertebrate ferritin, except for the N-terminal extension region, the C-terminal short helix E, and the end of the BC-loop. Similar to the crystal structures of other ferritins, metal binding sites were observed in the iron entry channel, ferroxidase center, and nucleation site of SFER4. In addition to these conventional sites, a novel metal binding site was discovered intermediate between the iron entry channel and the ferroxidase site. This site was coordinated by the acidic side chain of Glu(173) and carbonyl oxygen of Thr(168), which correspond, respectively, to Glu(140) and Thr(135) of human H chain ferritin according to their sequences. A comparison of the ferroxidase activities of the native and the E173A mutant of SFER4 clearly showed a delay in the iron oxidation rate of the mutant. This indicated that the glutamate residue functions as a transit site of iron from the 3-fold entry channel to the ferroxidase site, which may be universal among ferritins. PMID- 20007327 TI - Bayesian network analysis of targeting interactions in chromatin. AB - In eukaryotes, many chromatin proteins together regulate gene expression. Chromatin proteins often direct the genomic binding pattern of other chromatin proteins, for example, by recruitment or competition mechanisms. The network of such targeting interactions in chromatin is complex and still poorly understood. Based on genome-wide binding maps, we constructed a Bayesian network model of the targeting interactions among a broad set of 43 chromatin components in Drosophila cells. This model predicts many novel functional relationships. For example, we found that the homologous proteins HP1 and HP1C each target the heterochromatin protein HP3 to distinct sets of genes in a competitive manner. We also discovered a central role for the remodeling factor Brahma in the targeting of several DNA binding factors, including GAGA factor, JRA, and SU(VAR)3-7. Our network model provides a global view of the targeting interplay among dozens of chromatin components. PMID- 20007328 TI - Bacterial Hen1 is a 3' terminal RNA ribose 2'-O-methyltransferase component of a bacterial RNA repair cassette. AB - Hen1 is an RNA ribose 2'-O-methyltransferase that modifies the 3' terminal nucleoside of eukaryal small regulatory RNAs. Here, we report that Hen1 homologs are present in bacterial proteomes from eight different phyla. Bacterial Hen1 is encoded by the proximal ORF of a two-gene operon that also encodes polynucleotide kinase-phosphatase (Pnkp), an RNA repair enzyme. Purified recombinant Clostridium thermocellum Hen1 is a homodimer of a 465-amino acid polypeptide. CthHen1 catalyzes methyl transfer from AdoMet to the 3' terminal nucleoside of an RNA oligonucleotide, but is unreactive with a synonymous DNA oligonucleotide or an RNA with a single 3'-terminal deoxyribose sugar. CthHen1 is optimally active at alkaline pH and dependent on manganese. Activity is inhibited by AdoHcy and abolished by mutations D291A and D316A in the putative AdoMet-binding pocket. The C-terminal fragment, Hen1-(259-465), comprises an autonomous monomeric methyltransferase domain. PMID- 20007329 TI - Profiling non-lysyl tRNAs in HIV-1. AB - During its assembly, human HIV-1 selectively packages the tRNA(Lys) isoacceptors, including tRNA(Lys3), the primer for the reverse transcriptase. However, other low molecular weight RNA species are also seen in the virus. We profiled the tRNAs packaged into HIV-1 using microarray analysis and validated our results by two-dimensional gel electrophoresis and RT-PCR. In addition to tRNA(Lys) isoacceptors, tRNA(Asn) and the rare isoacceptor of tRNA(Ile) are also selectively packaged. In Gag viral-like particles missing the GagPol protein, overall tRNA incorporation is reduced by >80%. This reduction is significantly greater than can be accounted for by the reduction in tRNA(Lys) isoacceptors, tRNA(Asn) and tRNA(Ile), suggesting that incorporation of other tRNAs may also require the GagPol protein. These results demonstrate selective incorporation of non-lysyl tRNAs into HIV-1 and highlight the application of microarrays as a novel method to study tRNA incorporation into viruses. PMID- 20007326 TI - The secret life of NAD+: an old metabolite controlling new metabolic signaling pathways. AB - A century after the identification of a coenzymatic activity for NAD(+), NAD(+) metabolism has come into the spotlight again due to the potential therapeutic relevance of a set of enzymes whose activity is tightly regulated by the balance between the oxidized and reduced forms of this metabolite. In fact, the actions of NAD(+) have been extended from being an oxidoreductase cofactor for single enzymatic activities to acting as substrate for a wide range of proteins. These include NAD(+)-dependent protein deacetylases, poly(ADP-ribose) polymerases, and transcription factors that affect a large array of cellular functions. Through these effects, NAD(+) provides a direct link between the cellular redox status and the control of signaling and transcriptional events. Of particular interest within the metabolic/endocrine arena are the recent results, which indicate that the regulation of these NAD(+)-dependent pathways may have a major contribution to oxidative metabolism and life span extension. In this review, we will provide an integrated view on: 1) the pathways that control NAD(+) production and cycling, as well as its cellular compartmentalization; 2) the signaling and transcriptional pathways controlled by NAD(+); and 3) novel data that show how modulation of NAD(+)-producing and -consuming pathways have a major physiological impact and hold promise for the prevention and treatment of metabolic disease. PMID- 20007330 TI - An unexpected ending: noncanonical 3' end processing mechanisms. AB - Proper 3' end processing of a nascent transcript is critical for the functionality of the mature RNA. Although it has long been thought that virtually all long RNA polymerase II transcripts terminate in a poly(A) tail that is generated by endonucleolytic cleavage followed by polyadenylation, noncanonical 3' end processing mechanisms have recently been identified at several gene loci. Unexpectedly, enzymes with well-characterized roles in other RNA processing events, such as tRNA biogenesis and pre-mRNA splicing, cleave these nascent transcripts to generate their mature 3' ends despite the presence of nearby polyadenylation signals. In fact, the presence of multiple potential 3' end cleavage sites is the norm at many human genes, and recent work suggests that the choice among sites is regulated during development and in response to cellular cues. It is, therefore, becoming increasing clear that the selection of a proper 3' end cleavage site represents an important step in the regulation of gene expression and that the mature 3' ends of RNA polymerase II transcripts can be generated via multiple mechanisms. PMID- 20007332 TI - Neisseria meningitidis with reduced susceptibility to quinolones in Singapore. PMID- 20007331 TI - Integrase variability and susceptibility to HIV integrase inhibitors: impact of subtypes, antiretroviral experience and duration of HIV infection. AB - BACKGROUND: Little is known about the extent and predictors of polymorphisms potentially influencing the susceptibility to HIV integrase inhibitors (INIs). METHODS: Genetic sequences of HIV integrase were obtained from INI-naive patients at two European clinics. The 39 amino acid changes at 29 integrase positions so far associated with INI resistance were examined according to HIV clade, prior antiretroviral exposure and duration of HIV infection. RESULTS: Integrase sequences were obtained from 418 patients, 294 (70.3%) infected with clade B and 124 (29.7%) infected with non-B variants (predominantly CRF02, A, C and D). Overall, 40% of patients were antiretroviral experienced and 32.8% were recent seroconverters. The most prevalent INI resistance-associated mutations were V72I (63.9%), V201I (54.8%), T206S (25.4%), I203M (9.8%) and K156N (7.4%). Major INI resistance mutations at positions 66, 92, 143, 148 and 155 were not detected. The mean number of polymorphic sites was greater in non-B than in B variants (2.17 versus 1.59; P < 0.001), and in antiretroviral-experienced than in drug-naive patients (1.89 versus 1.68; P = 0.034), whereas no significant differences were seen comparing recent seroconverters and chronically infected persons. CONCLUSIONS: Major INI resistance-associated mutations are very rare, if indeed ever present, in INI-naive patients. However, polymorphisms at positions which may influence the genetic barrier and/or drive the selection of specific INI resistance pathways are common, especially in HIV non-B subtypes. PMID- 20007333 TI - Resistance profile of the new nucleoside reverse transcriptase inhibitor apricitabine. AB - Apricitabine is a novel deoxycytidine nucleoside reverse transcriptase inhibitor (NRTI) currently in clinical development for the treatment of HIV infection. Apricitabine shows antiviral activity in vitro against HIV-1 strains and clinical isolates with mutations in the reverse transcriptase that confer resistance to other NRTIs, including M184V, thymidine analogue mutations (TAMs), nucleoside associated mutations such as L74V and certain mutations at codon 69. Apricitabine has shown activity in treatment-experienced HIV-1-infected patients with NRTI resistance (with M184V and up to five TAMs) as well as in treatment-naive patients. Resistance to apricitabine is slow to develop in vitro and there has been little evidence of development of resistance to apricitabine in clinical use thus far, including patients receiving apricitabine for up to 48 weeks. The resistance profile of apricitabine suggests there is a low potential for cross resistance with the currently available NRTIs and, thus, apricitabine may provide a treatment option for treatment-experienced HIV-1-infected patients with resistance to other NRTIs. In particular, the activity of apricitabine in the presence of the M184V mutation, which confers high-level resistance to lamivudine and emtricitabine, lends it to being used as a replacement for deoxycytidine analogues in patients who have failed treatment with lamivudine or emtricitabine. PMID- 20007334 TI - Decreased serum TSH levels are not associated with mortality in the adult northeast German population. AB - OBJECTIVE: Results of cohort studies on the association between decreased serum TSH levels and mortality are conflicting. Some studies demonstrated an increased mortality risk in subjects with decreased serum TSH levels, others did not. Even meta-analyses revealed contradictory results. We undertook the present study to investigate the association between decreased serum TSH levels and mortality in the large population-based Study of Health in Pomerania (SHIP). DESIGN AND METHODS: Data from 3651 individuals from SHIP without known thyroid disorders or thyroid treatment were analyzed. Serum TSH, free triiodothyronine, and free thyroxine levels were determined by immunochemiluminescent procedures. Decreased TSH was defined as serum TSH levels below 0.25 mIU/l. Cox regression was used to associate decreased TSH levels with mortality. RESULTS: The median duration of follow-up was 8.5 years (30 126 person years). During follow-up, 299 individuals (6.9%) died corresponding to a death rate of 9.92 deaths per 1000 person years. Survival time was shorter in subjects with decreased serum TSH levels compared to euthyroid individuals. After adjustment for age and sex, however, there was no association between decreased serum TSH levels and all-cause mortality (hazard ratio: 0.95; 95% confidence interval: 0.67; 1.36). Likewise, decreased serum TSH levels were neither associated with cardiovascular nor with cancer mortality. CONCLUSIONS: There is no independent association of decreased serum TSH levels with all-cause, cardiovascular, and cancer mortality in the adult northeast German population. Although our study has some strengths, we cannot finally conclude on therapeutical implications in individuals with subclinical thyroid diseases. PMID- 20007335 TI - The polyunsaturated fatty acid composition of hepatic and plasma lipids differ by both sex and dietary fat intake in rats. AB - In rats and humans, females have higher liver and/or plasma docosahexaenoic acid (DHA) content than males. We hypothesized that the effect of variation in total fat or essential fatty acid intakes on liver and plasma fatty acid composition would differ between sexes. Rats were fed a low-fat soybean oil (LFS), high-fat soybean oil (HFS), or high-fat linseed oil (HFL) diet for 20 d. There were significant sex differences in LFS rats in proportions of (n-3) and (n-6) fatty acids in plasma and liver contingent on lipid class. Significant diet x sex interactions were observed for eicosapentaenoic acid (EPA), DHA, and arachidonic acid (AA) status. HFL females had a higher proportion of EPA in plasma and liver phosphatidylcholine (PC), DHA in liver triacylglycerol (TAG), and AA in plasma PC than HFS and LFS females. These findings show that the effect of varying dietary fat intake on (n-3) and (n-6) long-chain PUFA (LCPUFA) status is modified by sex. Liver phospholipid and TAG fatty acid product:substrate ratios suggested greater Delta6 desaturase (Delta6D) activity in females than in males. The HFL diet induced higher Delta6D mRNA expression compared with the LFS or HFS diets and HFL females had 10% higher expression of Delta6D mRNA than HFL males. Together, these findings show that sex is an important determinant of the effect of variations in fat and fatty acid intake on LCPUFA status, which may have implications for recommendations for fat and fatty acid intake in humans. PMID- 20007336 TI - Weight gain in the first two years of life is an important predictor of schooling outcomes in pooled analyses from five birth cohorts from low- and middle-income countries. AB - Schooling predicts better reproductive outcomes, better long-term health, and increased lifetime earnings. We used data from 5 cohorts (Brazil, Guatemala, India, the Philippines, and South Africa) to explore the relative importance of birthweight and postnatal weight gain for schooling in pooled analyses (n = 7945) that used appropriate statistical methods [conditional weight (CW) gain measures that are uncorrelated with prior weights] and controlled for confounding. One SD increase in birthweight, approximately 0.5 kg, was associated with 0.21 y more schooling and 8% decreased risk of grade failure. One SD increase in CW gain between 0 and 2 y, approximately 0.7 kg, was associated with higher estimates, 0.43 y more schooling, and 12% decreased risk of failure. One SD increase of CW gain between 2 and 4 y, approximately 0.9 kg, was associated with only 0.07 y more schooling but not with failure. Also, in children born in the lowest tertile of birthweight, 1 SD increase of CW between 0 and 2 y was associated with 0.52 y more schooling compared with 0.30 y in those in the upper tertile. Relationships with age at school entry were inconsistent. In conclusion, weight gain during the first 2 y of life had the strongest associations with schooling followed by birthweight; weight gain between 2 and 4 y had little relationship to schooling. Catch-up growth in smaller babies benefited schooling. Nutrition interventions aimed at women and children under 2 y are among the key strategies for achieving the millennium development goal of universal primary education by 2015. PMID- 20007337 TI - Consumption of a flaxseed-rich food is not more effective than a placebo in alleviating the climacteric symptoms of postmenopausal women. AB - Our objective in this work was to test the effects of daily intake of bread produced with partially defatted ground flaxseed on the climacteric symptoms and endometrial thickness of postmenopausal women. A double-blind, placebo controlled, randomized clinical trial was performed with 38 women who had been postmenopausal for 1-10 y and consumed 2 slices of bread containing 25 g of flaxseed (46 mg lignans) or wheat bran (<1 mg lignans; control) every day for 12 consecutive weeks. The outcome variables were the daily number of hot flashes, the Kupperman Menopausal Index (KMI), and endometrial thickness. The plasma lipid profile (total cholesterol and HDL, LDL, and VLDL cholesterol fractions and triglycerides) and the hormones estradiol, follicle-stimulating hormone, thyroid stimulating hormone, and free thyroxine also were measured. Food intake was evaluated by means of 2 24-h recalls, before and after the treatment. Twenty patients in the study group and 18 in the control group completed the study. The general characteristics did not differ between the 2 groups at the start of the study. Both had significant, but similar, reductions in hot flashes and KMI after 3 mo of treatment. Moreover, endometrial thickness was not affected in either group. Our findings clearly show that although flaxseed is safe, its consumption at this level (46 mg lignans/d) is no more effective than placebo for reducing hot flashes and KMI. PMID- 20007338 TI - Nondairy creamer, but not milk, delays the appearance of coffee phenolic acid equivalents in human plasma. AB - Chlorogenic acids (CGA) are antioxidants found in coffee. They are becoming of interest for their health-promoting effects, but bioavailability in humans is not well understood. We hypothesized that adding whole milk or sugar and nondairy creamer to instant coffee might modulate the bioavailability of coffee phenolics. Nine healthy participants were asked to randomly drink, in a crossover design, instant coffee (Coffee); instant coffee and 10% whole milk (Milk); or instant coffee, sugar, and nondairy creamer already premixed (Sugar/NDC). All 3 treatments provided the same amount of total CGA (332 mg). Blood was collected for 12 h after ingestion and plasma samples treated using a liquid-liquid extraction method that included a full enzymatic cleavage to hydrolyze all CGA and conjugates into phenolic acid equivalents. Hence, we focused our liquid chromatography-Electrospray ionization-tandem MS detection and quantification on caffeic acid (CA), ferulic acid (FA), and isoferulic acid (iFA) equivalents. Compared with a regular black instant coffee, the addition of milk did not significantly alter the area under the curve (AUC), maximum plasma concentration (C(max)), or the time needed to reach C(max) (T(max)). The C(max) of CA and iFA were significantly lower and the T(max) of FA and iFA significantly longer for the Sugar/NDC group than for the Coffee group. However, the AUC did not significantly differ. As a conclusion, adding whole milk did not alter the overall bioavailability of coffee phenolic acids, whereas sugar and nondairy creamer affected the T(max) and C(max) but not the appearance of coffee phenolics in plasma. PMID- 20007339 TI - A new simulation model estimates micronutrient levels to include in fortified blended foods used in food aid programs. AB - Current micronutrient levels in Public Law 480 fortified blended foods (FBF) may not be appropriate for all food aid beneficiaries, particularly infants and/or young children and pregnant and/or lactating women. A simulation model was developed to determine the micronutrient fortification levels to include in FBF for food aid programs with the goal of reducing the risk of inadequate micronutrient intakes without exceeding the tolerable upper intake level (UL) for any recipient group. For each micronutrient, the age and gender group with the highest daily Recommended Nutrient Intake (RNI) relative to energy requirement was identified and the effect of providing different percentages of that RNI (66, 75, and 100%) was simulated. In this modeling exercise, we also examined consumption of the FBF at 25 (the usual level), 50, and 100% of daily energy requirement. Results indicated that 2 FBF products are needed: a complementary food for age 6-36 mo and a supplementary food for the older groups. Both of the FBF could be fortified to supply at least 75% of the RNI to all groups, without exceeding the UL for most nutrients, if consumed at 25% of the energy requirement. Even if consumed at 50% of energy requirements, mean intakes of most micronutrients would not exceed the UL, although at 100% of the energy requirement, several micronutrients were undesirably high. We conclude that fortifying an FBF to provide 75% of the RNI would be appropriate for most micronutrients, but this level of fortification would not be appropriate for long term consumption of the FBF at 100% of the energy requirements. PMID- 20007340 TI - Fluid dynamics of cytotoxic safety cabinets. AB - This study investigated the specific fluid dynamics characteristics of cytotoxic safety cabinets (CSC), particularly those used in cancer drug reconstitution operations. Measurements taken on site were used to derive characteristic data for these cabinets. An in-depth laboratory investigation of airflows inside another CSC was also conducted. Anemometric values recorded on these two installations enabled the experimental validation of computational fluid dynamics methods applied to CSC. The digital flow simulations conducted provide a better understanding of the detailed flow structure inside a CSC and made it possible to study the influence of different operating parameters on the air velocity distribution inside the cabinet front opening: recycled air temperature, product protection airflow rate, suction openings spatial distribution, air compensation mode and draughts, operator arm penetration, and operator presence in front of the cabinet. PMID- 20007341 TI - Respirable dust measurements in the brick manufacturing industry. PMID- 20007342 TI - Automatic sample production by depositing solutions on filters for the organization of proficiency tests. AB - This article describes a device intended to produce replicas on filters by liquid deposition of anion or metal solutions. Schematically, the filters are housed in cassettes labelled automatically by means of a code. An automatic arm takes each cassette, reads the code, and deposits the amount of element required. Weighing before and after deposition allows the amount deposited to be accurately checked and determined. This automated system allows the production of replicas with high deposition regularity, replica dispersion for the most part being <1%. The samples produced can be used during proficiency tests where the assigned value is determined either by the participants or by the organizer. PMID- 20007343 TI - A possible catalytic role for NH4+ in Na+ reabsorption across the thick ascending limb. PMID- 20007344 TI - Corticosteroid-induced kidney dysmorphogenesis is associated with deregulated expression of known cystogenic molecules, as well as Indian hedgehog. AB - An intact genome is essential for kidney growth and differentiation, but less is known about whether, and how, an altered fetal milieu modifies these processes. Maternal low-protein diets perturb growth of the metanephros, the precursor of the mature kidney. Fetal corticosteroid overexposure may, in part, mediate this, because such diets downregulate placental 11beta-hydroxysteroid dehydrogenase-2, which degrades maternal corticosteroids. We report that glucocorticoid and mineralocorticoid receptors are expressed in mouse metanephric epithelia. Metanephroi maintained in organ culture with hydrocortisone (1.4 or 14 microM) underwent a dose-dependant deceleration of overall growth accompanied by cyst formation. Dexamethasone, a glucocorticoid, reproduced these outcomes, but aldosterone, a mineralocorticoid, did not. Hydrocortisone upregulated transcripts levels of cadherin-11 and downregulated prospero-related homeobox-1, hence mimicking reported effects of maternal low-protein diet. Hydrocortisone also upregulated transcripts encoding Na(+)-K(+)-ATPase subunits and ligands for the epidermal growth factor receptor, all previously implicated in renal cyst growth. The most upregulated transcript, however, was indian hedgehog, and the encoded protein was immunodetected in metanephric cysts. Furthermore, in the presence of hydrocortisone, cystogenesis, but not whole organ growth, was significantly reduced by cyclopamine, a drug downregulating hedgehog signaling. Finally, both glucocorticoid receptor and indian hedgehog proteins were detected by immunohistochemistry in cystic tubules within human dysplastic kidneys, consistent with the hypothesis that these molecules modify the severity of this congenital malformation. Collectively, our observations raise the possibility that enhanced hedgehog signaling is an important stimulus for renal cyst formation. Furthermore, pharmacological inhibition of this pathway should be explored as a potential therapy for renal cystic diseases, starting with relevant animal models. PMID- 20007345 TI - Short-term stimulation of the thiazide-sensitive Na+-Cl- cotransporter by vasopressin involves phosphorylation and membrane translocation. AB - Vasopressin influences salt and water transport in renal epithelia. This is coordinated by the combined action of V2 receptor-mediated effects along distinct nephron segments. Modulation of NaCl reabsorption by vasopressin has been established in the loop of Henle, but its role in the distal convoluted tubule (DCT), an effective site for fine regulation of urinary electrolyte composition and the target for thiazide diuretics, is largely unknown. The Na+-Cl- cotransporter (NCC) of DCT is activated by luminal trafficking and phosphorylation at conserved NH2-terminal residues. Here, we demonstrate the effects of short-term vasopressin administration (30 min) on NCC activation in Brattleboro rats with central diabetes insipidus (DI) using the V2 receptor agonist desmopressin (dDAVP). The fraction of NCC abundance in the luminal plasma membrane was significantly increased upon dDAVP as shown by confocal microscopy, immunogold cytochemistry, and Western blot, suggesting increased apical trafficking of the transporter. Changes were paralleled by augmented phosphorylation of NCC as detected by antibodies against phospho-threonine and phospho-serine residues (2.5-fold increase at Thr53 and 1.4-fold increase at Ser71). dDAVP-induced phosphorylation of NCC, studied in tubular suspensions in the absence of systemic effects, was enhanced as well (1.7-fold increase at Ser71), which points to the direct mode of action of vasopressin in DCT. Changes were more pronounced in early (DCT1) than in late DCT as distinguished by the distribution of 11beta-hydroxysteroid dehydrogenase 2 in DCT2. These results suggest that the vasopressin-V(2) receptor-NCC signaling cascade is a novel effector system to adjust transepithelial NaCl reabsorption in DCT. PMID- 20007347 TI - Contribution of impaired Nrf2-Keap1 pathway to oxidative stress and inflammation in chronic renal failure. AB - Oxidative stress and inflammation are constant features and major mediators of progression of chronic kidney disease (CKD). Nuclear factor erythroid-2-related factor-2 (Nrf2) confers protection against tissue injury by orchestrating antioxidant and detoxification responses to oxidative and electrophilic stress. While sources of oxidative stress and inflammation in the remnant kidney have been extensively characterized, the effect of CKD on Nrf2 activation and expression of its downstream gene products is unknown and was investigated. Subgroups of male Sprague-Dawley rats were subjected to 5/6 nephrectomy or sham operation and observed for 6 or 12 wk. Kidneys were then harvested, and Nrf2 activity and its downstream target gene products (antioxidant and phase II enzymes) were assessed. In addition, key factors involved in promoting inflammation and oxidative stress were studied. In confirmation of earlier studies, rats with chronic renal failure exhibited increased lipid peroxidation, glutathione depletion, NF-kappaB activation, mononuclear cell infiltration, and upregulation of monocyte chemoattractant protein-1, NAD(P)H oxidase, cyclooxygenase-2, and 12-lipoxygenase in the remnant kidney pointing to oxidative stress and inflammation. Despite severe oxidative stress and inflammation, remnant kidney tissue Nrf2 activity (nuclear translocation) was mildly reduced at 6 wk and markedly reduced at 12 wk, whereas the Nrf2 repressor Keap1 was upregulated and the products of Nrf2 target genes [catalase, superoxide dismutase, glutathione peroxidase, heme oxygenase-1, NAD(P)H quinone oxidoreductase, and glutamate-cysteine ligase] were reduced or unchanged at 6 wk and significantly diminished at 12 wk. Thus oxidative stress and inflammation in the remnant kidney are compounded by conspicuous impairment of Nrf2 activation and consequent downregulation of the antioxidant enzymes. PMID- 20007346 TI - De novo expression of podocyte proteins in parietal epithelial cells during experimental glomerular disease. AB - Studies have shown that certain cells of the glomerular tuft begin to express proteins considered unique to other cell types upon injury. Little is known about the response of parietal epithelial cells (PEC) to injury. To determine whether PECs change their phenotype upon injury to also express proteins traditionally considered podocyte specific, the following four models of glomerular disease were studied: the transforming growth factor (TGF)-beta1 transgenic mouse model of global glomerulosclerosis, the adriamycin model of focal segmental glomerulosclerosis (FSGS), the anti-glomerular basement membrane (GBM) model of crescentic glomerulonephritis, and the passive Heymann nephritis model of membranous nephropathy. Double immunostaining was performed with antibodies to podocyte-specific proteins (synaptopodin and Wilms' tumor 1) and antibodies to PEC specific proteins (paired box gene 8 and claudin-1). No double staining was detected in normal mice. In contrast, the results showed a statistical increase in the number of cells attached to Bowman basement membrane that were double positive for both podocyte/PEC proteins in TGF-beta1 transgenic, anti-GBM, and membranous animals. Double-positive cells for both podocyte and PEC proteins were also statistically increased in the glomerular tuft in TGF-beta1 transgenic, anti GBM, and FSGS mice. These results are consistent with glomerular cells coexpressing podocyte and PEC proteins in experimental glomerular disease, but not under normal circumstances. PMID- 20007348 TI - Loss of clusterin expression worsens renal ischemia-reperfusion injury. AB - Prevention of ischemia-reperfusion injury (IRI) is a challenge in clinical care of the patients with kidney transplants or acute kidney injury, and understanding of the intrinsic mechanisms of resistance to injury in the kidney will lead to a novel therapy. Clusterin, a secreted glycoprotein, is an antiapoptotic protein in cancer cells. Our study is to investigate the role of clusterin in renal IRI. Renal IRI in mice was induced by clamping renal vein and artery for 45 or 50 min at 32 degrees C. Apoptosis of renal tubular epithelial cells (TECs) was determined by FACS analysis. Clusterin expression was examined by Western blot or immunohistochemistry. Here, we showed that clusterin protein was induced in TECs following IRI, and more tubules expressed clusterin in the kidneys following ischemia at higher temperatures. In human proximal TEC HKC-8 cultures, clusterin was upregulated by removal of serum and growth factors in medium and was downregulated by TNF-alpha-IFN-gamma mixture. The levels of clusterin were positively correlated with cell survival in these conditions. Knockdown or knockout of clusterin expression enhanced the sensitivity of TECs to apoptosis. In experimental models of renal IRI, deficiency in clusterin expression worsened the injury, as indicated by a significant increase in renal tissue damage with higher levels of serum creatinine and blood urea nitrogen and by a poorer recovery from the injury in clusterin-deficient mice compared with wild-type mice. Our data indicate that the reduction of inducible expression of clusterin results in an increase in TEC apoptosis in the cultures and renders mice susceptibility to IRI, implying a protective role of clusterin in kidney injury. PMID- 20007349 TI - Potential involvement of P2Y2 receptor in diuresis of postobstructive uropathy in rats. AB - AVP resistance of the medullary collecting duct (mCD) in postobstructive uropathy (POU) has been attributed to increased production of PGE2. P2Y2 receptor activation causes production of PGE2 by the mCD. We hypothesize that increased P2Y2 receptor expression and/or activity may contribute to the diuresis of POU. Sprague-Dawley rats were subjected to bilateral ureteral obstruction for 24 h followed by release (BUO/R, n = 17) or sham operation (SHM/O, n = 15) and euthanized after 1 wk or 12 days. BUO/R rats developed significant polydipsia, polyuria, urinary concentration defect, and increased urinary PGE2 and decreased aquaporin-2 protein abundance in the inner medulla compared with SHM/O rats. After BUO/R, the relative mRNA expression of P2Y2 and P2Y6 receptors was increased by 2.7- and 4.9-fold, respectively, without significant changes in mRNA expression of P2Y1 or P2Y4 receptor. This was associated with a significant 3.5 fold higher protein abundance of the P2Y2 receptor in BUO/R than SHM/O rats. When freshly isolated mCD fractions were challenged with different types of nucleotides (ATPgammaS, ADP, UTP, or UDP), BUO/R and SHM/O rats responded to only ATPgammaS and UTP and released PGE2, consistent with involvement of the P2Y2, but not P2Y6, receptor. ATPgammaS- or UTP-stimulated increases in PGE2 were much higher in BUO/R (3.20- and 2.28-fold, respectively, vs. vehicle controls) than SHM/O (1.68- and 1.30-fold, respectively, vs. vehicle controls) rats. In addition, there were significant 2.4- and 2.1-fold increases in relative mRNA expression of prostanoid EP1 and EP3 receptors, respectively, in the inner medulla of BUO/R vs. SHM/O rats. Taken together, these data suggest that increased production of PGE2 by the mCD in POU may be due to increased expression and activity of the P2Y2 receptor. Increased mRNA expression of EP1 and EP3 receptors in POU may also help accentuate PGE2-induced signaling in the mCD. PMID- 20007350 TI - Comparative effect of direct renin inhibition and AT1R blockade on glomerular filtration barrier injury in the transgenic Ren2 rat. AB - Renin-angiotensin system (RAS) activation contributes to kidney injury through oxidative stress. Renin is the rate-limiting step in angiotensin (ANG II) generation. Recent work suggests renin inhibition improves proteinuria comparable to ANG type 1 receptor (AT1R) blockade (ARB). Thereby, we investigated the relative impact of treatment with a renin inhibitor vs. an ARB on renal oxidative stress and associated glomerular structural and functional changes in the transgenic Ren2 rat, which manifests hypertension, albuminuria, and increased tissue RAS activity. Young Ren2 and age-matched Sprague-Dawley (SD) controls (age 6-9 wk) were treated with a renin inhibitor (aliskiren), an ARB (irbesartan), or vehicle for 21 days. Ren2 rats exhibited increases in systolic pressure (SBP), albuminuria, and renal 3-nitrotyrosine content as well as ultrastructural podocyte foot-process effacement and diminution of the podocyte-specific protein nephrin. Structural and functional alterations were accompanied by increased renal cortical ANG II, AT1R, as well as NADPH oxidase subunit (Nox2) expression compared with SD controls. Abnormalities were attenuated to a similar extent with both aliskiren and irbesartan treatment. Despite the fact the dose of irbesartan used caused a greater reduction in SBP than aliskerin treatment (P < 0.05), the effects on proteinuria, nephrin, and oxidative stress were similar between the two treatments. Our results highlight both the importance of pressor-related reductions on podocyte integrity and albuminuria as well as RAS-mediated oxidant stress largely comparable between ARB and renin inhibition treatment. PMID- 20007351 TI - 8-pCPT-cGMP stimulates alphabetagamma-ENaC activity in oocytes as an external ligand requiring specific nucleotide moieties. AB - Epithelial sodium channels (ENaC) are regulated by protein kinase A, in addition to a broad spectrum of other protein kinases. It is not clear whether cGMP/PKG signaling might regulate ENaC activity. We examined the responses of alphabetagamma-ENaC channels expressed in Xenopus oocytes to 8-(4 chlorophenylthio)-cGMP (8-pCPT-cGMP), a cell-permeable cGMP analog. This compound stimulated human alphabetagamma-ENaC activity in a dose-dependent fashion, but cell-impermeable cGMP had no effect. Similar stimulatory effects of cGMP were observed in oocytes expressing either mouse or rat alphabetagamma-ENaC channels. The identical ion selectivity and amiloride sensitivity of the 8-pCPT-cGMP activated currents to those of alphabetagamma-ENaC channels suggest that the cGMP activated currents are associated with expressed ENaC. The PKGI activator Sp isomer of beta-phenyl-1,N(2)-etheno-8-bromo-cGMP did not elicit a rise in ENaC current and that the 8-pCPT-cGMP-induced activation of ENaC channels was blocked by incubating oocytes with a PKG inhibitor, but not with other cGMP-sensitive kinase inactivators for PKA, MEK, MAP, and PKC. Surprisingly, both site-directed mutation of putative consensus PKG phosphorylation sites and truncation of entire cytosolic NH(2)- and COOH-terminal tails did not alter the response to 8-pCPT cGMP. The ENaC activity was activated to the same extent by 8-pCPT-cGMP in cells in which PKGII expression was knocked down using small interfering RNA. Analog to 8-CPT-cAMP, 8-pCPT-cGMP was capable of activating ENaC in the identical manner in cell-free outside-out patches. We conclude that the rapid upregulation of human alphabetagamma-ENaC activity in oocytes by external 8-pCPT-cGMP and 4 chlorothiolphenol-cAMP depends on the para-chlorophenylthiol and the hydroxy groups, and 8-pCPT-cGMP may serve as a novel ENaC ligand in addition to activating PKG signal. PMID- 20007352 TI - Renal vascular dysfunction precedes the development of renal damage in the hypertensive Fawn-Hooded rat. AB - It is unknown whether generalized vascular dysfunction precedes the development of kidney disease. Therefore, we studied myogenic constriction and endothelium mediated dilatory responses in two inbred Fawn-Hooded (FH) rat strains, one of which spontaneously develops hypertension, proteinuria, and glomerulosclerosis (FHH), whereas the other (FHL) does not. Small renal, mesenteric resistance arteries and thoracic aorta isolated from FH rats before (7 wk old) and after the development of mild proteinuria (12 wks old) were mounted in perfused and isometric set-ups, respectively. Myogenic response, endothelium-dependent relaxation, and the contribution of endothelium-mediated dilatory compounds were studied using their respective inhibitors. Myogenic reactivity was assessed constructing pressure-diameter curves in the presence and absence of calcium. At the age of 7 wk, renal arteries isolated from kidneys of FHH rats developed significantly lower myogenic tone compared with FHL, most likely because of excessive cyclo-oxygenase 1-mediated production of constrictive prostaglandins. Consequently, young FHH demonstrated reduced maximal myogenic tone (22 +/- 4.8 vs. 10.8 +/- 2.0%, P = 0.03) and the peak myogenic index (-6.9 +/- 4.8 vs. 0.6 +/ 0.8%/mmHg, P = 0.07 for FHL vs. FHH, respectively). Active myogenic curves obtained in mesenteric arteries isolated from 7-wk-old rats did not differ between either strain, demonstrating a similar level of systemic myogenic tone in FHL and FHH rats. Therefore, before any renal end-organ damage is present, myogenic response seems selectively impaired in renal vasculature of FHH rats. Aortic reactivity did not differ between FHL and FHH at the time points studied. The present study shows that vascular dysfunction in both small renal and systemic arteries precedes renal end-organ damage in a spontaneous model of hypertension-associated renal damage. These early vascular changes might be potentially involved in the increased susceptibility of FHH rats to renal injury. PMID- 20007353 TI - Alterations in receptor-binding properties of swine influenza viruses of the H1 subtype after isolation in embryonated chicken eggs. AB - Alterations of the receptor-binding properties of swine influenza A viruses (SIVs) during their isolation in embryonated chicken eggs have not been well studied. In this study, the receptor-binding properties of classical H1 SIVs isolated solely in eggs or Madin-Darby canine kidney (MDCK) cells were examined. Sequencing analysis revealed substitutions of D190V/N or D225G in the haemagglutinin (HA) proteins in egg isolates, whereas MDCK isolates retained HA genes identical to those of the original viruses present in the clinical samples. Egg isolates with substitution of either D190V/N or D225G had increased haemagglutinating activity for mouse and sheep erythrocytes, but reduced activity for rabbit erythrocytes. Additionally, egg isolates with D225G had increased haemagglutination activity for chicken erythrocytes. A direct binding assay using a sialyl glycopolymer that possessed either a 5-N-acetylneuraminic acid (Neu5Ac) alpha2,6galactose (Gal) or a Neu5Acalpha2,3Gal linkage revealed that the egg isolates used in this study showed higher binding activity to the Neu5Acalpha2,3Gal receptor than MDCK isolates. Increased binding activity of the egg isolates to the Neu5Acalpha2,3Gal receptor was also confirmed by haemagglutination assay with resialylated chicken erythrocytes by Galbeta1,3/4GlcNAcalpha2,3-sialyltransferase. These observations were reinforced by flow-cytometric and N-glycan analyses of the erythrocytes. The alpha2,3-linked sialic acids were expressed predominantly on the surface of mouse and sheep erythrocytes. Chicken erythrocytes expressed Neu5Acalpha2,3Gal more abundantly than Neu5Acalpha2,6Gal, and rabbit erythrocytes expressed both 5-N glycolylneuraminic acid (Neu5Gc) alpha2,6Gal and Neu5Acalpha2,6Gal. Our results demonstrate clearly that classical H1 SIVs undergo alterations in receptor binding activity associated with an amino acid substitution in the HA protein during isolation and propagation in embryonated chicken eggs. PMID- 20007354 TI - Human papillomavirus 5 and 8 E6 downregulate interleukin-8 secretion in primary human keratinocytes. AB - Human papillomaviruses (HPVs) of the genus Betapapillomavirus appear to be involved in the early stages of skin cancer development, since both the prevalence and viral load are higher in precancerous actinic keratoses than in skin cancers. Interleukin-8 (IL-8) is an inflammatory cytokine that serves to alert the surrounding tissue after UV-induced damage. We examined the effects of the E2, E6 and E7 proteins of HPV8 and the E6 proteins of various HPV genotypes on IL-8 secretion from primary keratinocytes. HPV5 and HPV8 E6 showed the highest downregulation of basal IL-8 secretion. HPV8 E6 also negatively modulated IL-8 mRNA expression and protein secretion upon UVB irradiation. The downregulation of IL-8 in actinic keratoses may weaken the response to UV-induced damage and thus favour the accumulation of UVB-induced mutations. PMID- 20007355 TI - Antibody to parvovirus B19 nonstructural protein is associated with chronic arthralgia in patients with chronic fatigue syndrome/myalgic encephalomyelitis. AB - Chronic fatigue syndrome/myalgic encephalomyelitis (CFS/ME) is a neuro-immune disease of uncertain pathogenesis. Human parvovirus B19 infection has been shown to occur just prior to development of the onset of CFS/ME in several cases, although B19 seroprevalence studies do not show any significant differences between CFS/ME and controls. In this study, we analysed parvovirus B19 markers in CFS/ME patients (n=200), diagnosed according to Fukuda CDC criteria, and normal blood donors (n=200). Serum from each subject was tested for anti-B19 VP2 IgM and IgG (by Biotrin ELISA), anti-B19 NS1 IgM and IgG (by immunofluorescence), and B19 DNA (by real-time PCR). CFS/ME patients and normal blood donors had a similar B19 seroprevalence (75 % versus 78 %, respectively). Eighty-three CFS patients (41.5 %) as compared with fourteen (7 %) normal blood donors tested positive for anti B19 NS1 IgG (chi(2)=64.8; P<0.0001; odds ratio=9.42, CI 5.11-17.38). Of these 83 patients, 61 complained of chronic joint pain, while 22 did not. Parvovirus B19 DNA was detected in serum of 11 CFS patients and none of the controls by Taqman real-time PCR (chi(2)=9.35, P<0.002). Positivity for anti-B19 NS1 IgG was associated with higher expression levels of the human CFS-associated genes NHLH1 and GABPA. As NS1 antibodies are thought to indicate chronic or severe courses of B19 infection, these findings suggest that although the seroprevalence of B19 in CFS patients is similar to controls, the immune control of the virus in these patients may not be efficient. PMID- 20007356 TI - An interfering activity against lymphocytic choriomeningitis virus replication associated with enhanced mutagenesis. AB - Previous studies have documented that, in the presence of the mutagenic base analogue 5-fluorouracil (FU), lymphocytic choriomeningitis virus (LCMV) that persisted in BHK-21 cells decreased its infectivity to a larger extent than intracellular viral RNA levels, prior to virus extinction. This observation, together with in silico simulations, led to the proposal of the lethal defection model of virus extinction. This model suggests the participation of defective interfering genomes in the loss of infectivity by increased mutagenesis. Since LCMV naturally produces defective-interfering particles, it was important to show that a capacity to interfere is produced in association with FU treatment. Here, we document that BHK-21 cells persistently infected with LCMV grown in the presence of FU, but not in its absence, generated an interfering activity that suppressed LCMV infectivity. Interference was specific for LCMV and was sensitive to UV irradiation and its activity was dose- and time-dependent. The interfering preparations produced positive LCMV immunofluorescence and viral particles seen by electron microscopy when used to infect cells, despite some preparations being devoid of detectable infectivity. Interference did not involve significant increases of mutant spectrum complexity, as predicted by the lethal defection model. The results provide support for a specific interference associated with LCMV when the virus replicates in the presence of FU. The excess of interference relative to that observed in the absence of FU is necessary for LCMV extinction. PMID- 20007357 TI - PB2 and PA genes control the expression of the temperature-sensitive phenotype of cold-adapted B/USSR/60/69 influenza master donor virus. AB - The cold-adapted (ca) and temperature-sensitive (ts) influenza master donor virus (MDV) B/USSR/60/69 was derived from its wild-type parental virus after successive passages in eggs at 32 degrees C and 25 degrees C. This strain is currently in use for preparing reassortant influenza B vaccine viruses which are used in the Russian trivalent live attenuated influenza vaccine. Vaccine viruses are obtained by classical reassortment of MDV and a currently circulating wild-type virus. The phenotypic properties cold adaptation and temperature sensitivity are inherited from the six genes encoding the internal proteins of the MDV. However, the role of the individual gene segments in temperature sensitivity and thus attenuation is not known. In this study, 35 reassortant viruses of B/USSR/60/69 MDV with current wild-type non-ts influenza B viruses were generated in eggs or MDCK cells and studied in order to identify the genes responsible for their ts phenotype. For each virus the exact genome composition was determined as well as its ts phenotype. The results demonstrated that the polymerase PB2 and PA gene segments of B/USSR/60/69 MDV independently controlled expression of the ts phenotype of B/USSR/60/69 MDV-based reassortant viruses. The other genes coding for internal proteins played no role in this respect. This suggests that mutations in the polymerase genes PB2 and PA play an essential role in attenuation of B/USSR/60/69 MDV-based reassortant influenza B vaccine viruses. PMID- 20007358 TI - Classical swine fever virus infection protects aortic endothelial cells from pIpC mediated apoptosis. AB - Classical swine fever virus (CSFV) causes severe disease in pigs associated with leukopenia, haemorrhage and fever. We show that CSFV infection protects endothelial cells from apoptosis induced by the dsRNA mimic, pIpC, but not from other apoptotic stimuli, FasL or staurosporine. CSFV infection inhibits pIpC induced caspase activation, mitochondrial membrane potential loss and cytochrome c release as well as the pro-apoptotic effects of truncated Bid (tBid) overexpression. The CSFV proteins N(pro) and E(rns) both contribute to CSFV inhibition of apoptosis. We conclude that CSFV infection can inhibit apoptotic signalling at multiple levels, including at the caspase-8 and the mitochondrial checkpoints. By supporting viral replication, endothelial cells may promote CSFV pathogenesis. PMID- 20007359 TI - Glial activation involvement in neuronal death by Japanese encephalitis virus infection. AB - Japanese encephalitis is characterized by profound neuronal destruction/dysfunction and concomitant microgliosis/astrogliosis. Although substantial activation of glia is observed in Japanese encephalitis virus (JEV) induced Japanese encephalitis, the inflammatory responses and consequences of astrocytes and microglial activation after JEV infection are not fully understood. In this study, infection of cultured neurons/glia with JEV caused neuronal death and glial activation, as evidenced by morphological transformation, increased cell proliferation and elevated tumour necrosis factor (TNF)-alpha, interleukin (IL)-1beta, IL-6 and RANTES (regulated upon activation, normal T-cell expressed and secreted) production. Replication-competent JEV caused all glial responses and neurotoxicity. However, replication-incompetent JEV lost these abilities, except for the ability to change microglial morphology. The bystander damage caused by activated glia also contributed to JEV-associated neurotoxicity. Microglia underwent morphological changes, increased cell proliferation and elevated TNF-alpha, IL-1beta, IL-6 and RANTES expression in response to JEV infection. In contrast, IL-6 and RANTES expression, but no apparent morphological changes, proliferation or TNF-alpha/IL-1beta expression, was demonstrated in JEV-infected astrocytes. Supernatants of JEV-infected microglia, but not JEV-infected astrocytes, induced glial activation and triggered neuronal death. Antibody neutralization studies revealed that TNF-alpha and IL-1beta, but not RANTES or IL-6, released by activated microglia appeared to play roles in JEV-associated neurotoxicity. In conclusion, following JEV infection, neuronal death was accompanied by concomitant microgliosis and astrogliosis, and neurotoxic mediators released by JEV-activated microglia, rather than by JEV-activated astrocytes, had the ability to amplify the microglial response and cause neuronal death. PMID- 20007360 TI - A soluble form of P74 can act as a per os infectivity factor to the Autographa californica multiple nucleopolyhedrovirus. AB - The baculovirus occlusion-derived virion (ODV) is required to spread virus infection among insect hosts via the per os route. The Autographa californica multicapsid nucleopolyhedrovirus P74 protein is an ODV envelope protein that is essential for ODVs to be infectious. P74 is anchored in the ODV envelope by a C terminal transmembrane anchor domain and is N-terminally exposed on the ODV surface. In the present study, a series of N-terminal and C-terminal truncation mutants of P74 were evaluated for their ability to rescue per os infectivity of the P74-null virus, AcLP4. It was discovered that a P74 truncation mutant lacking the C-terminal transmembrane anchor domain of P74 was able to rescue per os infection. This result shows that a soluble form of P74 retains per os infectivity factor function and suggests that P74 may be complexed with other proteins in the ODV envelope. PMID- 20007361 TI - Immune responses in cattle inoculated with Mycobacterium bovis, Mycobacterium tuberculosis, or Mycobacterium kansasii. AB - Cattle were inoculated with Mycobacterium bovis, Mycobacterium tuberculosis, or Mycobacterium kansasii to compare the antigen-specific immune responses to various patterns of mycobacterial disease. Disease expression ranged from colonization with associated pathology (M. bovis infection) and colonization without pathology (M. tuberculosis infection) to no colonization or pathology (M. kansasii infection). Delayed-type hypersensitivity and gamma interferon responses were elicited by each mycobacterial inoculation; however, the responses by the M. bovis- and M. tuberculosis-inoculated animals exceeded those of the M. kansasii inoculated animals. Specific antibody responses were detected in all M. tuberculosis- and M. bovis-inoculated cattle 3 weeks after inoculation. From 6 to 16 weeks after M. tuberculosis inoculation, the antibody responses waned, whereas the responses persisted with M. bovis infection. With M. kansasii inoculation, initial early antibody responses waned by 10 weeks after inoculation and then increased 2 weeks after the injection of purified protein derivative for the skin test at 18 weeks after challenge. These findings indicate that antibody responses are associated with the antigen burden rather than the pathology, cellular immune responses to tuberculin correlate with infection but not necessarily with the pathology or bacterial burden, and exposure to mycobacterial antigens may elicit an antibody response in a presensitized animal. PMID- 20007362 TI - Immunogenicity of a psoralen-inactivated dengue virus type 1 vaccine candidate in mice. AB - We evaluated a novel psoralen-inactivated dengue virus type 1 (DENV-1) vaccine candidate in Mus musculus mice. Mice received intradermal alum or 5 to 10 ng of psoralen-inactivated virus. Anti-DENV-1 neutralizing antibody was detectable in 10/11 mice receiving a 10-ng dose at 90 days. Psoralen-inactivated DENV-1 is immunogenic in mice. PMID- 20007363 TI - A live oral recombinant Salmonella enterica serovar typhimurium vaccine expressing Clostridium perfringens antigens confers protection against necrotic enteritis in broiler chickens. AB - Necrotic enteritis (NE) in broiler chickens is caused by Clostridium perfringens, and there is currently no effective vaccine for NE. We previously showed that in broiler chickens protection against NE can be achieved through intramuscular immunization with alpha toxin (AT) and hypothetical protein (HP), and we subsequently identified B-cell epitopes in HP. In the present study, we identified B-cell epitopes in AT recognized by chickens immune to NE. The gene fragments encoding immunodominant epitopes of AT as well as those of HP were codon optimized for Salmonella and cloned into pYA3493, and the resultant plasmid constructs were introduced into an attenuated Salmonella enterica serovar Typhimurium chi9352 vaccine vehicle. The expression of these Clostridium perfringens proteins, alpha toxoid (ATd) and truncated HP (HPt), was confirmed by immunoblotting. The protection of broiler chickens against experimentally induced NE was assessed at both the moderate and the severe levels of challenge. Birds immunized orally with Salmonella expressing ATd were significantly protected against moderate NE, and there was a nonsignificant trend for protection against severe challenge, whereas HPt-immunized birds were significantly protected against both severities of challenge. Immunized birds developed serum IgY and mucosal IgA and IgY antibody responses against Clostridium and Salmonella antigens. In conclusion, this study identified, for the first time, the B-cell epitopes in AT from an NE isolate recognized by chickens and showed the partial protective ability of codon-optimized ATd and HPt against NE in broiler chickens when they were delivered orally by using a Salmonella vaccine vehicle. PMID- 20007364 TI - Association of reduced tumor necrosis factor alpha, gamma interferon, and interleukin-1beta (IL-1beta) but increased IL-10 expression with improved chest radiography in patients with pulmonary tuberculosis. AB - Mycobacterium tuberculosis infection is a major world health issue. The early identification of patients at risk for a poor response to anti-M. tuberculosis therapy would help elucidate the key players in the anti-M. tuberculosis response. The objective of the present study was to correlate the modulation of cytokine expression (interleukin-1 [IL-1], IL-6, IL-8, IL-10, IL-12, gamma interferon [IFN-gamma], interferon-inducible protein [IP-10], and monocyte chemotactic protein 1 [MCP-1]) with the clinical response to 2 months of intensive therapy. From January to December 2007, 40 M. tuberculosis-infected patients and 40 healthy patients were recruited. After exclusion for diabetes, 32 patients and 36 controls were analyzed. The clinical responses of the M. tuberculosis-infected patients on the basis of the findings of chest radiography were compared to their plasma cytokine levels measured before and after 2 months of intensive anti-M. tuberculosis therapy and 6 months of therapy with human cytokine antibody arrays. Chest radiographs of 20 of 32 M. tuberculosis-infected patients showed improvement after 2 months of intensive therapy (early responders), while the M. tuberculosis infections in 12 of 32 of the patients resolved after a further 4 months (late responders). The levels of expression of TNF-alpha, MCP-1, IFN-gamma, and IL-1beta were decreased; and the level of IL-10 increased in early responders. After adjustment for age, gender, and the result of sputum culture for M. tuberculosis, significant differences in the levels of MCP-1 and IP-10 expression were observed between the early and the late responders after 2 months of intensive anti-M. tuberculosis therapy. Due to the interpatient variability in IP-10 levels, intrapatient monitoring of IP-10 levels may provide more insight into the M. tuberculosis responder status than comparison between patients. Plasma MCP-1 levels were normalized in patients who had resolved their M. tuberculosis infections. Further studies to evaluate the association of the modulation in MCP-1 levels with early and late responses are warranted. PMID- 20007365 TI - Diagnosis of Chikungunya fever in an Indian population by an indirect enzyme linked immunosorbent assay protocol based on an antigen detection assay: a prospective cohort study. AB - A Chikungunya virus (CHIKV) outbreak continues in India. Monitoring of the clinical features of CHIKV infection is an important component of assessing the disease process. Diagnosis is usually made by an immunoglobulin M (IgM)/IgG enzyme-linked immunosorbent assay (ELISA). However, these assays have extremely low sensitivities for the detection of infection in the majority of CHIKV patients during the acute stage of infection (during the 1 to 4 days after infection). In our laboratory, a sensitive ELISA protocol for antigen detection has been developed for the detection of CHIKV infection in the acute stage, and in the present study we assessed the usefulness of this ELISA-based system for the detection of CHIKV infection. We performed a prospective, double-blinded study of 205 Indian patients with suspected CHIKV infection in the Nagpur District. All patients underwent a full clinical assessment, and their serum samples were analyzed for the presence of antigens and of IgM and IgG by an ELISA protocol. In patients with CHIKV infection, the sensitivity of antigen detection was 85%, which was significantly higher (P < 0.001) than that of IgM (17%) or IgG (45%) detection. The sensitivity of IgM (20%) or IgG (25%) detection was significantly lower than that of the antigen assay (95%) for patients with acute infections (i.e., from day 1 to day 5 after infection). Antigen detection not only gives a positive confirmatory result in the early phase of the disease, but it is also useful in the prodromal and subclinical stage and may be useful for field applications for the rapid detection of CHIKV infection. PMID- 20007366 TI - Evaluation of three multiplex flow immunoassays compared to an enzyme immunoassay for the detection and differentiation of IgG class antibodies to herpes simplex virus types 1 and 2. AB - The diagnosis of herpes simplex virus (HSV) infections is routinely made based on clinical findings and supported by laboratory testing using PCR or viral culture. However, in instances of subclinical or unrecognized HSV infection, serologic testing for IgG class antibodies to type-specific HSV glycoprotein G (gG) may be useful. This study evaluated and compared the performances of three multiplex flow immunoassays (AtheNA Multi-Lyte [Zeus Scientific], BioPlex 2200 [Bio-Rad Laboratories], and Plexus HerpeSelect [Focus Diagnostics]) for the simultaneous detection of gG type-specific IgG antibodies to HSV types 1 and 2 (HSV-1 and HSV 2). Serum specimens (n = 505) submitted for routine gG type-specific HSV IgG testing by enzyme immunoassay (EIA) (HerpeSelect; Focus Diagnostics) were also tested by the three multiplex flow immunoassays. Specimens showing discordant results were tested by HSV type-specific Western blotting (WB). For HSV-1 IgG, the AtheNA, BioPlex, and Plexus assays demonstrated agreements of 94.9% (479/505 specimens), 97.8% (494/505 specimens), and 97.4% (492/505 specimens), respectively, with the results of EIA. For HSV-2 IgG, the AtheNA, BioPlex, and Plexus assays showed agreements of 87.9% (444/505 specimens), 97.2% (491/505 specimens), and 96.8% (489/505 specimens), respectively, with EIA results. Timing studies showed that the AtheNA, BioPlex, and Plexus assays could provide complete analysis of 90 serum specimens in 3.1, 1.5, and 2.9 h, respectively, versus 3.1 h by EIA. These findings suggest that the gG type-specific HSV IgG multiplex immunoassays may be beneficial to high-volume clinical laboratories experiencing significant increases in the number of specimens submitted for HSV serologic testing. The evaluated systems provide comparable results to those of EIA, while reducing hands-on time and eliminating the necessity to aliquot specimens prior to testing. PMID- 20007367 TI - Plant extinctions and introductions lead to phylogenetic and taxonomic homogenization of the European flora. AB - Human activities have altered the composition of biotas through two fundamental processes: native extinctions and alien introductions. Both processes affect the taxonomic (i.e., species identity) and phylogenetic (i.e., species evolutionary history) structure of species assemblages. However, it is not known what the relative magnitude of these effects is at large spatial scales. Here we analyze the large-scale effects of plant extinctions and introductions on taxonomic and phylogenetic diversity of floras across Europe, using data from 23 regions. Considering both native losses and alien additions in concert reveals that plant invasions since AD 1500 exceeded extinctions, resulting in (i) increased taxonomic diversity (i.e., species richness) but decreased phylogenetic diversity within European regions, and (ii) increased taxonomic and phylogenetic similarity among European regions. Those extinct species were phylogenetically and taxonomically unique and typical of individual regions, and extinctions usually were not continent-wide and therefore led to differentiation. By contrast, because introduced alien species tended to be closely related to native species, the floristic differentiation due to species extinction was lessened by taxonomic and phylogenetic homogenization effects. This was especially due to species that are alien to a region but native to other parts of Europe. As a result, floras of many European regions have partly lost and will continue to lose their uniqueness. The results suggest that biodiversity needs to be assessed in terms of both species taxonomic and phylogenetic identity, but the latter is rarely used as a metric of the biodiversity dynamics. PMID- 20007368 TI - A genome-wide screen in Saccharomyces cerevisiae reveals a critical role for the mitochondria in the toxicity of a trichothecene mycotoxin. AB - Trichothecene mycotoxins synthesized by Fusarium species are potent inhibitors of eukaryotic translation. They are encountered in both the environment and in food, posing a threat to human and animal health. They have diverse roles in the cell that are not limited to the inhibition of protein synthesis. To understand the trichothecene mechanism of action, we screened the yeast knockout library to identify genes whose deletion confers resistance to trichothecin (Tcin). The largest group of resistant strains affected mitochondrial function, suggesting a role for fully active mitochondria in trichothecene toxicity. Tcin inhibited mitochondrial translation in the wild-type strain to a greater extent than in the most resistant strains, implicating mitochondrial translation as a previously unrecognized site of action. The Tcin-resistant strains were cross-resistant to anisomycin and chloramphenicol, suggesting that Tcin targets the peptidyltransferase center of mitochondrial ribosomes. Tcin-induced cell death was partially rescued by mutants that regulate mitochondrial fusion and maintenance of the tubular morphology of mitochondria. Treatment of yeast cells with Tcin led to the fragmentation of the tubular mitochondrial network, supporting a role for Tcin in disruption of mitochondrial membrane morphology. These results provide genome-wide insight into the mode of action of trichothecene mycotoxins and uncover a critical role for mitochondrial translation and membrane maintenance in their toxicity. PMID- 20007369 TI - Giant Marseillevirus highlights the role of amoebae as a melting pot in emergence of chimeric microorganisms. AB - Giant viruses such as Mimivirus isolated from amoeba found in aquatic habitats show biological sophistication comparable to that of simple cellular life forms and seem to evolve by similar mechanisms, including extensive gene duplication and horizontal gene transfer (HGT), possibly in part through a viral parasite, the virophage. We report here the isolation of "Marseille" virus, a previously uncharacterized giant virus of amoeba. The virions of Marseillevirus encompass a 368-kb genome, a minimum of 49 proteins, and some messenger RNAs. Phylogenetic analysis of core genes indicates that Marseillevirus is the prototype of a family of nucleocytoplasmic large DNA viruses (NCLDV) of eukaryotes. The genome repertoire of the virus is composed of typical NCLDV core genes and genes apparently obtained from eukaryotic hosts and their parasites or symbionts, both bacterial and viral. We propose that amoebae are "melting pots" of microbial evolution where diverse forms emerge, including giant viruses with complex gene repertoires of various origins. PMID- 20007370 TI - FM dyes enter via a store-operated calcium channel and modify calcium signaling of cultured astrocytes. AB - The amphiphilic fluorescent styryl pyridinium dyes FM1-43 and FM4-64 are used to probe activity-dependent synaptic vesicle cycling in neurons. Cultured astrocytes can internalize FM1-43 and FM4-64 inside vesicles but their uptake is insensitive to the elevation of cytosolic calcium (Ca(2+)) concentration and the underlying mechanism remains unclear. Here we used total internal reflection fluorescence microscopy and pharmacological tools to study the mechanisms of FM4-64 uptake into cultured astrocytes from mouse neocortex. Our data show that: (i) endocytosis is not a major route for FM4-64 uptake into astrocytes; (ii) FM4-64 enters astrocytes through an aqueous pore and strongly affects Ca(2+) homeostasis; (iii) partitioning of FM4-64 into the outer leaflet of the plasma membrane results in a facilitation of store-operated Ca(2+) entry (SOCE) channel gating; (iv) FM4-64 permeates and competes with Ca(2+) for entry through a SOCE channel; (v) intracellular FM4-64 mobilizes Ca(2+) from the endoplasmic reticulum stores, conveying a positive feedback to activate SOCE and to sustain dye uptake into astrocytes. Our study demonstrates that FM dyes are not markers of cycling vesicles in astrocytes and calls for a careful interpretation of FM fluorescence. PMID- 20007371 TI - Chromogranin B P413L variant as risk factor and modifier of disease onset for amyotrophic lateral sclerosis. AB - Recently, chromogranins were reported to interact specifically with mutant forms of superoxide dismutase that are linked to amyotrophic lateral sclerosis (ALS). This interaction led us to analyze the frequencies of sequence variants of the CHGB gene in ALS patients and matched controls from three different countries. Of particular interest was the finding of the P413L CHGB variant present in 10% of ALS patients (n = 705) as compared to 4.5% in controls (n = 751), conferring a 2.2-fold greater relative risk to develop the disease (P < 0.0001). This effect was mainly contributed by the samples of French origin that yielded a frequency of the P413L variation at 17% in ALS (n = 289) and 5% in controls (n = 448), conferring a 3.3-fold greater risk to develop ALS. Furthermore, the P413L CHGB variant is associated with an earlier age of onset by almost a decade in both sporadic ALS and familial ALS cases. Genetic variation influencing age of onset in ALS had not previously been reported. Expression of fusion CHGB-EGFP constructs in SHSY-5Y cells revealed that the P413L variation can cause defective sorting of CHGB into secretory granules. The finding that CHGB may act as a susceptibility gene and modifier of onset in ALS is consistent with the emerging view that dysfunction of the secretory pathway may contribute to increased vulnerability of motor neurons. PMID- 20007372 TI - Loss of MeCP2 in aminergic neurons causes cell-autonomous defects in neurotransmitter synthesis and specific behavioral abnormalities. AB - Rett syndrome (RTT) is characterized by specific motor, cognitive, and behavioral deficits. Because several of these abnormalities occur in other disease states associated with alterations in aminergic neurotransmitters, we investigated the contribution of such alterations to RTT pathogenesis. We found that both individuals with RTT and Mecp2-null mice have lower-than-normal levels of aminergic metabolites and content. Deleting Mecp2 from either TH-positive dopaminergic and noradrenergic neurons or PET1-positive serotonergic neurons in mice decreased corresponding neurotransmitter concentration and specific phenotypes, likely through MeCP2 regulation of rate-limiting enzymes involved in aminergic neurotransmitter production. These data support a cell-autonomous, MeCP2-dependent mechanism for the regulation of aminergic neurotransmitter synthesis contributing to unique behavioral phenotypes. PMID- 20007373 TI - The impact of long-distance horizontal gene transfer on prokaryotic genome size. AB - Horizontal gene transfer (HGT) is one of the most dominant forces molding prokaryotic gene repertoires. These repertoires can be as small as approximately 200 genes in intracellular organisms or as large as approximately 9,000 genes in large, free-living bacteria. In this article we ask what is the impact of HGT from phylogenetically distant sources, relative to the size of the gene repertoire. Using different approaches for HGT detection and focusing on both cumulative and recent evolutionary histories, we find a surprising pattern of nonlinear enrichment of long-distance transfers in large genomes. Moreover, we find a strong positive correlation between the sizes of the donor and recipient genomes. Our results also show that distant horizontal transfers are biased toward those functional groups that are enriched in large genomes, showing that the trends in functional gene content and the impact of distant transfers are interdependent. These results highlight the intimate relationship between environmental and genomic complexity in microbes and suggest that an ecological, as opposed to phylogenetic, signal in gene content gains relative importance in large-genomed bacteria. PMID- 20007374 TI - A nitric oxide/cysteine interaction mediates the activation of soluble guanylate cyclase. AB - Nitric oxide (NO) regulates a number of essential physiological processes by activating soluble guanylate cyclase (sGC) to produce the second messenger cGMP. The mechanism of NO sensing was previously thought to result exclusively from NO binding to the sGC heme; however, recent studies indicate that heme-bound NO only partially activates sGC and additional NO is involved in the mechanism of maximal NO activation. Furthermore, thiol oxidation of sGC cysteines results in the loss of enzyme activity. Herein the role of cysteines in NO-stimulated sGC activity investigated. We find that the thiol modifying reagent methyl methanethiosulfonate specifically inhibits NO activation of sGC by blocking a non heme site, which defines a role for sGC cysteine(s) in mediating NO binding. The nature of the NO/cysteine interaction was probed by examining the effects of redox active reagents on NO-stimulated activity. These results show that NO binding to, and dissociation from, the critical cysteine(s) does not involve a change in the thiol redox state. Evidence is provided for non-heme NO in the physiological activation of sGC in context of a primary cell culture of human umbilical vein endothelial cells. These findings have relevance to diseases involving the NO/cGMP signaling pathway. PMID- 20007375 TI - Temporal self-organization of the cyclin/Cdk network driving the mammalian cell cycle. AB - We propose an integrated computational model for the network of cyclin-dependent kinases (Cdks) that controls the dynamics of the mammalian cell cycle. The model contains four Cdk modules regulated by reversible phosphorylation, Cdk inhibitors, and protein synthesis or degradation. Growth factors (GFs) trigger the transition from a quiescent, stable steady state to self-sustained oscillations in the Cdk network. These oscillations correspond to the repetitive, transient activation of cyclin D/Cdk4-6 in G(1), cyclin E/Cdk2 at the G(1)/S transition, cyclin A/Cdk2 in S and at the S/G(2) transition, and cyclin B/Cdk1 at the G(2)/M transition. The model accounts for the following major properties of the mammalian cell cycle: (i) repetitive cell cycling in the presence of suprathreshold amounts of GF; (ii) control of cell-cycle progression by the balance between antagonistic effects of the tumor suppressor retinoblastoma protein (pRB) and the transcription factor E2F; and (iii) existence of a restriction point in G(1), beyond which completion of the cell cycle becomes independent of GF. The model also accounts for endoreplication. Incorporating the DNA replication checkpoint mediated by kinases ATR and Chk1 slows down the dynamics of the cell cycle without altering its oscillatory nature and leads to better separation of the S and M phases. The model for the mammalian cell cycle shows how the regulatory structure of the Cdk network results in its temporal self-organization, leading to the repetitive, sequential activation of the four Cdk modules that brings about the orderly progression along cell-cycle phases. PMID- 20007376 TI - Using a functional enzyme model to understand the chemistry behind hydrogen sulfide induced hibernation. AB - The toxic gas H(2)S is produced by enzymes in the body. At moderate concentrations, H(2)S elicits physiological effects similar to hibernation. Herein, we describe experiments that imply that the phenomenon probably results from reversible inhibition of the enzyme cytochrome c oxidase (CcO), which reduces oxygen during respiration. A functional model of the oxygen-reducing site in CcO was used to explore the effects of H(2)S during respiration. Spectroscopic analyses showed that the model binds two molecules of H2S. The electro-catalytic reduction of oxygen is reversibly inhibited by H(2)S concentrations similar to those that induce hibernation. This phenomenon derives from a weak, reversible binding of H(2)S to the Fe(II) porphyrin, which mimics heme a(3) in CcO's active site. No inhibition of CcO is detected at lower H(2)S concentrations. Nevertheless, at lower concentrations, H(2)S could have other biological effects on CcO. For example, H(2)S rapidly reduces Fe(III) and Cu(II) in both the oxidized form of this functional model and in CcO itself. H(2)S also reduces CcO's biological reductant, cytochrome c, which normally derives its reducing equivalents from food metabolism. Consequently, it is speculated that H(2)S might also serve as a source of electrons during periods of hibernation when food supplies are low. PMID- 20007377 TI - Campbell's monkeys concatenate vocalizations into context-specific call sequences. AB - Primate vocal behavior is often considered irrelevant in modeling human language evolution, mainly because of the caller's limited vocal control and apparent lack of intentional signaling. Here, we present the results of a long-term study on Campbell's monkeys, which has revealed an unrivaled degree of vocal complexity. Adult males produced six different loud call types, which they combined into various sequences in highly context-specific ways. We found stereotyped sequences that were strongly associated with cohesion and travel, falling trees, neighboring groups, nonpredatory animals, unspecific predatory threat, and specific predator classes. Within the responses to predators, we found that crowned eagles triggered four and leopards three different sequences, depending on how the caller learned about their presence. Callers followed a number of principles when concatenating sequences, such as nonrandom transition probabilities of call types, addition of specific calls into an existing sequence to form a different one, or recombination of two sequences to form a third one. We conclude that these primates have overcome some of the constraints of limited vocal control by combinatorial organization. As the different sequences were so tightly linked to specific external events, the Campbell's monkey call system may be the most complex example of 'proto-syntax' in animal communication known to date. PMID- 20007378 TI - Structural analysis of the catalytically inactive kinase domain of the human EGF receptor 3. AB - The kinase domain of human epidermal growth factor receptor (HER) 3/ErbB3, a member of the EGF receptor (EGFR) family, lacks several residues that are critical for catalysis. Because catalytic activity in EGFR family members is switched on by an allosteric interaction between kinase domains in an asymmetric kinase domain dimer, HER3 might be specialized to serve as an activator of other EGFR family members. We have determined the crystal structure of the HER3 kinase domain and show that it appears to be locked into an inactive conformation that resembles that of EGFR and HER4. Although the crystal structure shows that the HER3 kinase domain binds ATP, we confirm that it is catalytically inactive but can serve as an activator of the EGFR kinase domain. The HER3 kinase domain forms a dimer in the crystal, mediated by hydrophobic contacts between the N-terminal lobes of the kinase domains. This N-lobe dimer closely resembles a dimer formed by inactive HER4 kinase domains in crystal structures determined previously, and molecular dynamics simulations suggest that the HER3 and HER4 N-lobe dimers are stable. The kinase domains of HER3 and HER4 form similar chains in their respective crystal lattices, in which N-lobe dimers are linked together by reciprocal exchange of C-terminal tails. The conservation of this tiling pattern in HER3 and HER4, which is the closest evolutionary homolog of HER3, might represent a general mechanism by which this branch of the HER receptors restricts ligand-independent formation of active heterodimers with other members of the EGFR family. PMID- 20007379 TI - Revising the recent evolutionary history of equids using ancient DNA. AB - The rich fossil record of the family Equidae (Mammalia: Perissodactyla) over the past 55 MY has made it an icon for the patterns and processes of macroevolution. Despite this, many aspects of equid phylogenetic relationships and taxonomy remain unresolved. Recent genetic analyses of extinct equids have revealed unexpected evolutionary patterns and a need for major revisions at the generic, subgeneric, and species levels. To investigate this issue we examine 35 ancient equid specimens from four geographic regions (South America, Europe, Southwest Asia, and South Africa), of which 22 delivered 87-688 bp of reproducible aDNA mitochondrial sequence. Phylogenetic analyses support a major revision of the recent evolutionary history of equids and reveal two new species, a South American hippidion and a descendant of a basal lineage potentially related to Middle Pleistocene equids. Sequences from specimens assigned to the giant extinct Cape zebra, Equus capensis, formed a separate clade within the modern plain zebra species, a phenotypicically plastic group that also included the extinct quagga. In addition, we revise the currently recognized extinction times for two hemione related equid groups. However, it is apparent that the current dataset cannot solve all of the taxonomic and phylogenetic questions relevant to the evolution of Equus. In light of these findings, we propose a rapid DNA barcoding approach to evaluate the taxonomic status of the many Late Pleistocene fossil Equidae species that have been described from purely morphological analyses. PMID- 20007380 TI - Correlation of genotype and in vitro susceptibilities of Cryptococcus gattii strains from the Pacific Northwest of the United States. AB - Cryptococcus gattii emerged in North America in 1999 as a human and veterinary pathogen on Vancouver Island, British Columbia. The emergent subtype, VGIIa, and the closely related subtype VGIIb can now be found in the United States in Washington, Oregon, and California. We performed multilocus sequence typing and antifungal susceptibility testing on 43 isolates of C. gattii from human patients in Oregon, Washington, California, and Idaho. In contrast to Vancouver Island, VGIIa was the most frequent but not the predominant subtype in the northwest United States. Antifungal susceptibility testing showed statistically significant differences in MICs between the subtypes. This is the first study to apply antifungal susceptibility testing to C. gattii isolates from the Pacific Northwest and the first to make direct comparisons between subtypes. PMID- 20007381 TI - K2 serotype Klebsiella pneumoniae causing a liver abscess associated with infective endocarditis. AB - Klebsiella pneumoniae primary liver abscess (KPLA) is an emerging disease that is associated with distant septic complications. We report the first case of KPLA associated with infective endocarditis. The K. pneumoniae strain was a hypermucoid K2 serotype carrying the rmpA virulence-associated gene. PMID- 20007382 TI - Characterization of Streptococcus milleri group isolates from expectorated sputum of adult patients with cystic fibrosis. AB - With the recent insights into the Streptococcus milleri group (SMG) as pulmonary pathogens in patients with cystic fibrosis (CF), we sought to characterize 128 isolates from the sputum of adults with CF, along with 45 isolates from patients with invasive diseases for comparison. The tests performed included Lancefield grouping; tests for hemolysis; tests for the production of hyaluronidase, chondroitin sulfatase, DNase, proteases, and hydrogen peroxide; and PCR for the detection of the intermedilysin gene (ily). We also generated biochemical profiles with the Rapid ID Strep 32 API system and tested cell-free supernatants for the presence of the signal molecule autoinducer-2 (AI-2) using a Vibrio harveyi bioassay with a subset of CF strains. The S. intermedius isolates from both strain collections were similar, while the S. constellatus and S. anginosus isolates yielded several biotypes that differed in prevalence between the two strain collections. Beta-hemolytic, Lancefield group C S. constellatus comprised 74.4% of the S. constellatus isolates from patients with CF but only 13.3% of the corresponding isolates from patients with invasive infections. This was the only S. constellatus biotype associated with pulmonary exacerbations. Hyaluronidase positive S. anginosus was detected only among the isolates from patients with CF. Strain-to-strain variability in AI-2 expression was evident, with the mean values being the highest for S. anginosus, followed by S. constellatus and then S. intermedius. Cluster analysis and 16S rRNA sequencing revealed that the species of SMG could be accurately determined with a minimum of three phenotypic tests: tests for the Lancefield group, hyaluronidase production, and chondroitin sulfatase production. Furthermore, isolates from patients with invasive infections clustered with isolates from the sputum of patients with CF, suggesting that the respiratory tract isolates were equally pathogenic. PMID- 20007383 TI - Detecting VIM-1 production in Proteus mirabilis by an imipenem-dipicolinic acid double disk synergy test. PMID- 20007384 TI - Serotyping pneumococcal meningitis cases in the African meningitis belt by use of multiplex PCR with cerebrospinal fluid. AB - We reformulated a multiplex PCR algorithm for serotyping of pneumococcal meningitis directly on cerebrospinal fluid (CSF). Compared to established methods on isolates, CSF-based PCR had at least 80% sensitivity and 100% specificity. In regional meningitis surveillance, CSF-based PCR increased the serotype information yield from 40% of cases (isolate testing) to 90%. PMID- 20007385 TI - PCR using blood for diagnosis of invasive pneumococcal disease: systematic review and meta-analysis. AB - The use of molecular-based methods for the diagnosis of bacterial infections in blood is appealing, but they have not yet passed the threshold for clinical practice. A systematic review of prospective and case-control studies assessing the diagnostic utility of PCR directly with blood samples for the diagnosis of invasive pneumococcal disease (IPD) was performed. A broad search was conducted to identify published and unpublished studies. Two reviewers independently extracted the data. Summary estimates for sensitivity and specificity with 95% confidence intervals (CIs) were calculated by using the hierarchical summary receiver operating characteristic method. The effects of sample processing, PCR type, the gene-specific primer, study design, the participants' age, and the source of infection on the diagnostic odds ratios were assessed through meta regression. Twenty-nine studies published between 1993 and 2009 were included. By using pneumococcal bacteremia for case definition and healthy people or patients with bacteremia caused by other bacteria as controls (22 studies), the summary estimates for sensitivity and specificity were 57.1% (95% CI, 45.7 to 67.8%) and 98.6% (95% CI, 96.4 to 99.5%), respectively. When the controls were patients suspected of having IPD without pneumococcal bacteremia (26 studies), the respective values were 66.4% (95% CI, 55.9 to 75.6%) and 87.8% (95% CI, 79.5 to 93.1%). With lower degrees of proof for IPD (any culture or serology result and the clinical impression), the sensitivity of PCR decreased and the specificity increased. All analyses were highly heterogeneous. The use of nested PCR and being a child were associated with low specificity, while the use of a cohort study design was associated with a low sensitivity. The lack of an appropriate reference standard might have caused underestimation of the performance of the PCR. Currently available methods for PCR with blood samples for the diagnosis of IPD lack the sensitivity and specificity necessary for clinical practice. PMID- 20007386 TI - Carriage of community-associated methicillin-resistant Staphylococcus aureus in a cohort of infants in southern Israel: risk factors and molecular features. AB - There are few data about the epidemiology of community-associated methicillin resistant Staphylococcus aureus (CA-MRSA) among children in Israel. This study was intended to identify risk factors for CA-MRSA colonization in healthy infants, to characterize the molecular features of colonizing organisms, and to determine whether they are responsible for health care-associated (HA) infections. Nasal cultures and demographic details were collected from a cohort of healthy infants at 5 visits between the ages of 2 and 12 months. Clinical characteristics of pediatric MRSA bloodstream infections (2001 to 2006) and wound cultures collected over 6 months were also studied. Clonal structure was evaluated by multilocus sequence typing. Isolates were studied for the staphylococcal cassette chromosome mec (SCCmec) type and for the presence of Panton-Valentine leukocidin (PVL) genes. MRSA was cultured at least once from 45 of 659 infants (346 Jewish and 313 Bedouin infants). Forty of 45 (89%) isolates were from Bedouin infants. Twenty-nine of 45 (64.4%) belonged to a new clonal complex, designated CC913, that carries SCCmec IV but not the PVL genes. CC913 was also isolated from 9/14 blood cultures and 7/8 wounds. All CC913 infections occurred in Bedouin children, and all but two were HA. In conclusion, Bedouin origin was the main risk factor for carriage of CA-MRSA. CC913 was dominant both in healthy carriers and as a cause of pediatric HA-MRSA bloodstream infections. PMID- 20007387 TI - Optimal threshold for a positive hybrid capture 2 test for detection of human papillomavirus: data from the ARTISTIC trial. AB - We present data on the use of the Hybrid Capture 2 (HC2) test for the detection of high-risk human papillomavirus (HR HPV) with different thresholds for positivity within a primary screening setting and as a method of triage for low grade cytology. In the ARTISTIC population-based trial, 18,386 women were screened by cytology and for HPV. Cervical intraepithelial neoplasia lesions of grade two and higher (CIN2+ lesions) were identified for 453 women within 30 months of an abnormal baseline sample. When a relative light unit/cutoff (RLU/Co) ratio of > or = 1 was used as the threshold for considering an HC2 result positive, 15.6% of results were positive, and the proportion of CIN2+ lesions in this group was 14.7%. The relative sensitivity for CIN2+ lesion detection was 93.4%. When an RLU/Co ratio of > or = 2 was used as the threshold, there was a 2.5% reduction in positivity, with an increase in the proportion of CIN2+ lesions detected. The relative sensitivity decreased slightly, to 90.3%. Among women with low-grade cytology, HPV prevalences were 43.7% and 40.3% at RLU/Co ratios of > or = 1 and > or = 2, respectively. The proportions of CIN2+ lesions detected were 17.3% and 18.0%, with relative sensitivities of 87.7% at an RLU/Co ratio of > or = 1 and 84.2% at an RLU/Co ratio of > or = 2. At an RLU/Co ratio of > or = 1, 68.3% of HC2-positive results were confirmed by the Roche line blot assay, compared to 77.2% of those at an RLU/Co ratio of > or = 2. Fewer HC2-positive results were confirmed for 35- to 64-year-olds (50.3% at an RLU/Co ratio of > or = 1 and 63.2% at an RLU/Co ratio of > 2) than for 20- to 34-year-olds (78.7% at an RLU/Co ratio of > or = 1 and 83.7% at an RLU/Co ratio of > 2). If the HC2 test is used for routine screening as an initial test or as a method of triage for low grade cytology, we would suggest increasing the threshold for positivity from the RLU/Co ratio of > or = 1, recommended by the manufacturer, to an RLU/Co ratio of > or = 2, since this study has shown that a beneficial balance between relative sensitivity and the proportion of CIN2+ lesions detected is achieved at this threshold. PMID- 20007388 TI - Clinical value of Treponema pallidum real-time PCR for diagnosis of syphilis. AB - The diagnosis of syphilis can be complicated when it is based on diverse clinical manifestations, dark-field microscopy, and serology. In the present study, therefore, we examined the additional clinical value of a Treponema pallidum real time TaqMan PCR for the detection of primary and secondary syphilis. The additional value of the T. pallidum real-time PCR for the diagnosis of primary syphilis was evaluated by the use of three different algorithms: (i) a head-to head comparison of the dark-field microscopy result and the T. pallidum real-time PCR result, (ii) comparison of the clinical diagnosis made in a sexually transmitted infection clinic (STI) (including by dark-field microscopy) and the T. pallidum real-time PCR result, and (iii) comparison of the clinical diagnosis made in a general practitioner's office (without dark-field microscopy) and the T. pallidum real-time PCR result. A fourth algorithm was used to determine the performance of the T. pallidum real-time PCR regarding the detection of secondary syphilis. From December 2006 to April 2008, 716 patients with suspected cases of primary syphilis and 133 patients with suspected cases of secondary syphilis were included in the study. A kappa value of 0.601 was found for the agreement between dark-field microscopy and the T. pallidum real-time PCR. Good agreement was found between the T. pallidum real-time PCR and both the diagnosis of the general practitioner (kappa = 0.745) and the diagnosis of the STI clinic (kappa = 0.769). The sensitivity with respect to the STI clinic diagnosis was 72.8%, the specificity was 95.5%, the positive predictive value was 89.2%, and the negative predictive value was 95.0%. The T. pallidum real-time PCR is a fast, efficient, and reliable test for the diagnosis of primary syphilis in an STI outpatient clinic and a general practitioner setting, but it has no added diagnostic value for the diagnosis of secondary syphilis. PMID- 20007389 TI - Caspofungin Etest endpoint for Aspergillus isolates shows poor agreement with the reference minimum effective concentration. AB - The Clinical and Laboratory Standards Institute (CLSI) M38-A2 reference broth microdilution (BMD) method for the antifungal susceptibility testing of filamentous fungi now includes guidelines for testing echinocandin activity using the minimum effective concentration (MEC) as the endpoint measurement. In this study, we compared the caspofungin Etest MIC on RPMI agar and Mueller-Hinton agar (supplemented with glucose and methylene blue [MGM]) to the BMD MEC for 345 clinical Aspergillus isolates, including A. flavus, A. fumigatus, A. nidulans, A. niger, and A. terreus. The essential agreement (+/-1 log(2) dilution) of the Etest on MGM and RPMI agar with the reference BMD MEC was 18 and 26%, respectively. The geometric mean values for BMD MEC and MGM Etest were 0.137 and 0.024 microg/ml, respectively, and the geometric mean values for BMD and RPMI agar were 0.128 and 0.031 microg/ml, respectively. Comparatively, 91% of paired MGM and RPMI Etest results were within 2 log(2) dilutions of each other and consistently produced clearly defined endpoints. In conclusion, the caspofungin Etest MIC, like the BMD MEC, is a reproducible endpoint but is markedly lower than the reference BMD. In anticipation of susceptibility breakpoint assignments, optimization studies will be required to improve the concordance of these two assays so that the potential for underreporting echinocandin resistance in Aspergillus is mitigated. PMID- 20007390 TI - Wound botulism complicating internal fixation of a complex radial fracture. AB - Botulism developed in a patient following surgical repair of an open radial fracture. Symptoms resolved after treatment with antitoxin and antibiotics, and hardware excision was deferred. Subsequent osteomyelitis necessitated hardware exchange, and wound cultures grew Clostridium argentinense. This case highlights the management of botulism associated with orthopedic hardware. PMID- 20007391 TI - Morphologic and phylogenetic characterization of Conidiobolus lamprauges recovered from infected sheep. AB - Conidiobolus lamprauges, a soil and plant entomophthoralean fungus, has been reported only in a horse and, more recently, in sheep with rhinopharyngeal entomophthoramycosis. Thus, little information is available to enable proper identification of this pathogen and its differentiation from other saprotrophic and pathogenic Conidiobolus species. Using classical mycological tools and molecular methodologies, we report for the first time the taxonomic and phylogenetic description of three C. lamprauges isolates recovered from sheep with rhinopharyngeal entomophthoramycosis. The distinctive clinical and pathological features of C. lamprauges are compared with those of other Conidiobolus spp. affecting sheep, as well as with those of the stramenopilan ovine agent Pythium insidiosum. The comparative morphological attributes of Conidiobolus spp. are also diagramed; along with the sequence data generated, they should assist laboratories in the identification of these uncommon species. PMID- 20007392 TI - Outbreak of skin infections in college football team members due to an unusual strain of community-acquired methicillin-susceptible Staphylococcus aureus. AB - We report a skin and soft-tissue infection outbreak among football team members due to a USA300 methicillin-susceptible Staphylococcus aureus (MRSA) strain with genes coding for Panton-Valentine leukocidin and the arginine catabolic mobile element. We postulate that the strain is a community-associated USA300 MRSA strain that lost methicillin resistance but retained important virulence factors. PMID- 20007394 TI - Comparison of the Abbott RealTime CT new formulation assay with two other commercial assays for detection of wild-type and new variant strains of Chlamydia trachomatis. AB - In an analytical-method comparison study of clinical samples, the Abbott RealTime CT new formulation assay (m2000 real-time PCR), consisting of a duplex PCR targeting different parts of the cryptic plasmid in Chlamydia trachomatis, was compared both with version 2 of the Roche Cobas TaqMan CT assay, comprising a duplex PCR for a target in the cryptic plasmid and the omp1 gene, and with the Gen-Probe Aptima Combo 2 assay (AC2) targeting the C. trachomatis 23S rRNA molecule. First-catch urine samples from Sweden were tested in Malmo, Sweden, for C. trachomatis with the m2000 real-time PCR assay and with an in-house PCR for the new variant C. trachomatis strain with a deletion in the cryptic plasmid. Aliquots of the urine samples were sent to Aarhus, Denmark, where they were further examined with the TaqMan CT and AC2 assays. A positive prevalence of 9.1% (148/1,632 urine samples examined) was detected according to the combined reference standard. The sensitivities and specificities of the three assays were as follows: for the Abbott m2000 assay, 95.3% (141/148) and 99.9% (1,483/1,485), respectively; for the Roche TaqMan assay, 82.4% (122/148) and 100.0% (1,485/1,485); and for the Gen-Probe AC2 assay, 99.3% (147/148) and 99.9% (1,484/1,485). The plasmid mutant strain was detected in 24% (36/148) of the C. trachomatis-positive samples. There is a difference in sensitivity between the new formulations of the Abbott and the Roche assays, but both assays detected the wild-type and new variant C. trachomatis strains equally well. PMID- 20007393 TI - Integrated detection of extended-spectrum-beta-lactam resistance by DNA microarray-based genotyping of TEM, SHV, and CTX-M genes. AB - Extended-spectrum beta-lactamases (ESBL) of the TEM, SHV, or CTX-M type confer resistance to beta-lactam antibiotics in gram-negative bacteria. The activity of these enzymes against beta-lactam antibiotics and their resistance against inhibitors can be influenced by genetic variation at the single-nucleotide level. Here, we describe the development and validation of an oligonucleotide microarray for the rapid identification of ESBLs in gram-negative bacteria by simultaneously genotyping bla(TEM), bla(SHV), and bla(CTX-M). The array consists of 618 probes that cover mutations responsible for 156 amino acid substitutions. As this comprises unprecedented genotyping coverage, the ESBL array has a high potential for epidemiological studies and infection control. With an assay time of 5 h, the ESBL microarray also could be an attractive option for the development of rapid antimicrobial resistance tests in the future. The validity of the DNA microarray was demonstrated with 60 blinded clinical isolates, which were collected during clinical routines. Fifty-eight of them were characterized phenotypically as ESBL producers. The chip was characterized with regard to its resolution, phenotype genotype correlation, and ability to resolve mixed genotypes. ESBL phenotypes could be correctly ascribed to ESBL variants of bla(CTX-M) (76%), bla(SHV) (22%), or both (2%), whereas no ESBL variant of bla(TEM) was found. The most prevalent ESBLs identified were CTX-M-15 (57%) and SHV-12 (18%). PMID- 20007395 TI - Direct serogrouping of Streptococcus pneumoniae strains in clinical samples by use of a latex agglutination test. AB - Pneumotest-Latex (Statens Seruminstitut) was evaluated for direct serogrouping of Streptococcus pneumoniae strains in clinical samples from patients with invasive disease. The technique was accurate to its level of discrimination for 62 of 67 clinical samples (92.5%). Pneumotest-Latex would be a useful alternative for direct serogrouping of pneumococci in clinical samples. PMID- 20007396 TI - Unique finding of a 2009 H1N1 influenza virus-positive clinical sample suggests matrix gene sequence variation. PMID- 20007397 TI - Two-step scheme for rapid identification and differentiation of Legionella pneumophila and non-Legionella pneumophila species. AB - A rapid two-step scheme based on PCR amplification and enzymatic digestion analysis of a 226-bp fragment of the 16S rRNA gene was developed to identify the Legionella genus by PCR amplification and to differentiate the Legionella pneumophila and non-Legionella pneumophila species by enzymatic digestion analysis. Among 42 ATCC strains (16 strains of L. pneumophila and 26 strains of non-L. pneumophila) and 200 Legionella isolates from environmental water samples, including pools, rivers, lakes, and cooling towers in Guangdong province, 99.59% of L. pneumophila and non-L. pneumophila strains were correctly identified and differentiated by this scheme. The procedure of this two-step identification and differentiation scheme is simple and takes only about 4 h. These results suggest that this two-step scheme provides a simple and convenient method for the rapid identification and differentiation of L. pneumophila and non-L. pneumophila species. PMID- 20007398 TI - Selection and counterselection of the rtI233V adefovir resistance mutation during antiviral therapy. AB - Recently, we reported on three patients with chronic hepatitis B virus (HBV) infection for whom adefovir (ADF) therapy virologically failed, most likely due to a preexisting rtI233V HBV polymerase mutation. Here, we describe two further patients with chronic HBV infection who were found to develop the rtI233V mutation after initiation of ADF therapy. These patients represent the first cases known so far in which the rtI233V ADF resistance mutation evolved under persistent HBV replication during HBV therapy with ADF. Interestingly, one of the previously described patients, who was initially successfully switched from ADF to tenofovir (TDF) and became virologically suppressed subsequently, experienced a moderate but remarkable rebound of HBV viremia after switching from TDF to entecavir, due to the emergence of renal toxicity. Thus, we provide evidence for the selection and counterselection of the rtI233V ADF resistance mutation during antiviral therapy. PMID- 20007399 TI - Role of rapid immunochromatographic antigen testing in diagnosis of influenza A virus 2009 H1N1 infection. AB - Rapid antigen testing using immunochromatographic devices has become a diagnostic mainstay for detection of influenza virus and respiratory syncytial virus, the two major viruses infecting the respiratory tract. Recent studies have indicated that poor performance in the detection of the novel influenza A virus 2009 H1N1 should preclude their use. A survey of influenza diagnostic methods available on ClinMicroNet and Division C, the two ASM list servers, revealed that, despite this reported poor performance, a majority of the laboratories surveyed intend to continue to offer this testing during the current influenza season. Our two experts have been asked to consider the following question: what is the role of rapid immunochromatographic antigen testing in the laboratory diagnosis of influenza A virus infection during the current 2009 H1N1 pandemic? PMID- 20007400 TI - Repetitive extragenic palindromic PCR for study of Streptococcus mutans diversity and transmission in human populations. AB - Pulsed-field gel electrophoresis (PFGE) is considered the "gold standard" for molecular epidemiological study. Repetitive extragenic palindromic PCR (rep-PCR) is less time-consuming and more suitable for analyzing large numbers of bacterial strains in human populations. PFGE and rep-PCR provide comparable genotyping results for investigating Streptococcus mutans diversity and transmission. PMID- 20007401 TI - Comparative activity of pradofloxacin, enrofloxacin, and azithromycin against Bartonella henselae isolates collected from cats and a human. AB - Using Bartonella henselae isolates from cats and a human, the activity of pradofloxacin was compared with those of enrofloxacin and azithromycin. By Etest and disc diffusion assay, pradofloxacin showed greater antimicrobial activity than did other antibiotics. We conclude that pradofloxacin may prove useful for the treatment of B. henselae infections. PMID- 20007402 TI - First autochthonous case of Rhinocladiella mackenziei cerebral abscess outside the Middle East. AB - Cerebral phaeohyphomycosis due to Rhinocladiella mackenziei is a severe infection in the middle east, resulting in nearly 100% mortality despite the application of combined surgical and antifungal therapy and occurring occasionally in otherwise healthy patients. We report the first case of brain infection in a middle-aged male in India, where R. mackenziei is not endemic. PMID- 20007403 TI - Evaluation of risk factors for coinfection or cocolonization with vancomycin resistant enterococcus and methicillin-resistant Staphylococcus aureus. AB - We retrospectively evaluated 410 patients with coinfection or cocolonization due to vancomycin-resistant (VR) enterococcus (VRE) and methicillin-resistant Staphylococcus aureus (MRSA). The prevalence rate was 19.8%. Risk factors included isolation of VR Enterococcus faecalis and use of linezolid or clindamycin. Inc18-like vanA plasmids were found in 7% of VR E. faecalis isolates and none of the VR E. faecium isolates. PMID- 20007404 TI - Extracellular signal-regulated kinase 1/2 and c-Jun NH2-terminal kinase, through nuclear factor-kappaB and activator protein-1, contribute to caerulein-induced expression of substance P and neurokinin-1 receptors in pancreatic acinar cells. AB - The neuropeptide substance P (SP) has emerged to be an important proinflammatory mediator in acute pancreatitis (AP). The presence of substance P and its receptor, neurokinin-1 receptor (NK1R) has been shown in the pancreas and the pancreatic acinar cells. In this study, we investigated the unexplored mechanisms that mediate SP and NK1R expression using an in vitro AP model. Pancreatic acinar cells were obtained from pancreas of male Swiss mice. Isolated cells were treated with caerulein to mimic secretagogue pancreatitis. A concentration-dependent study that subjected the cells to 60 min of stimulation by caerulein showed that SP and the transcript from its gene preprotachykinin-A (PPT-A), and NK1R were up regulated at a supraphysiological concentration of 10(-7) M. A concentration dependent study on intracellular kinases, extracellular signal-regulated kinase (ERK1/2), and c-Jun N-terminal kinase (JNK) and also transcription factors nuclear factor-kappaB (NF-kappaB) and activator protein-1 (AP-1) showed that they were activated when the caerulein concentration was 10(-7) M. Inhibition of JNK reversed the up-regulation of PPT-A, SP, and NK1R. However, inhibition of ERK1/2 reversed the up-regulation of NK1R but not of PPT-A and SP. Furthermore, we found that specific ERK1/2 and JNK inhibitors reduce NF-kappaB and AP-1 activity. Taken together, our results suggest that supraphysiological concentrations of caerulein up-regulate the expression of SP and NK1R in pancreatic acinar cells, and the signaling molecules that are involved in this up-regulation include ERK1/2, JNK, NF-kappaB, and AP-1. PMID- 20007405 TI - Naturally occurring variants of human CBR3 alter anthracycline in vitro metabolism. AB - Doxorubicin (DOX) and daunorubicin (DAUN) are anthracycline anticancer agents; however, considerable interpatient variability exists in their pharmacokinetics. This interpatient variability is attributed in part to altered metabolism by nonsynonymous single-nucleotide polymorphisms (ns-SNPs) in genes encoding the carbonyl reductases. This study examines the effect of seven naturally occurring ns-SNPs in the CBR3 gene on in vitro metabolism of anthracyclines to doxorubicinol and daunorubicinol. Kinetic assays measure metabolite levels by high-performance liquid chromatography separation with fluorescence detection by use of purified, histidine-tagged, human CBR3 wild type and variant enzymes. The V224M, C4Y, and V93I variants resulted in significantly reduced maximal reaction velocity (V(max)) for both anthracyclines compared with the wild-type enzyme, whereas the M235L variant had significantly reduced V(max) for DOX only. Significant increases in substrate affinity were found for the V244M variant with DAUN, as well as the C4Y and V93I variants with DOX. The catalytic efficiency values for the V244M, C4Y, and V93I variants were significantly lower than the wild type for DAUN and DOX. Furthermore, DOX was observed to be a better substrate than DAUN for the wild-type enzyme and its variants. HapMap analysis indicated that a haplotype carrying the C4Y and V244M mutations may occur in some individuals in the 11 ethnic populations studied in the HapMap project. Our preparation of the double mutant indicated a significant reduction in activity compared with the wild-type enzyme and single-mutant preparations. These findings suggest that commonly occurring ns-SNPs in human CBR3 significantly alter the in vitro metabolism of DOX and DAUN. PMID- 20007407 TI - Interpreting sexually transmissible infection prevention trials by adjusting for the magnitude of exposure. AB - BACKGROUND: Exposures to sexually transmitted infections are discrete identifiable events. Interventions to prevent sexually transmitted infections have a certain probability of effectiveness in reducing risk in any given event. PURPOSE: Randomized control trials for sexually transmitted infections interventions are designed to estimate the effectiveness in preventing acquisition of infection. Typically, randomized control trials are run over a specific period of time and incidence in the control arm is compared with incidence in an intervention arm. However, it is possible that the effectiveness of an intervention over the duration of a clinical trial may be different to the actual effectiveness of the intervention in every single exposure event or the overall effectiveness over different periods than the duration of the trial. METHODS: In this study a simple mathematical framework is used, similar to methods in conception research, to describe the expected effectiveness that would be observed in a clinical trial of an intervention per-exposure and for clinically relevant shorter and longer durations than the trial, where each subject has multiple risk exposures. RESULTS: It is theoretically demonstrated that the actual effectiveness of the intervention per risk event is not equal to the overall preventative effectiveness of the intervention in preventing transmission over many exposures. Examples are given for sexually transmitted infections with diverse transmission probabilities (HIV and HPV) and for interventions with different levels of effectiveness (condoms and circumcision). The observed effectiveness of an intervention is likely to be maintained over many exposures for infections with low transmission risk (like HIV) but the observed effectiveness decreases substantially with number of exposures for moderate or high risk infections (like HPV). An equation is provided for interpreting randomized control trials' estimates of effectiveness with respect to various degrees of risk exposure. LIMITATIONS: The difficulty in adjusting the interpretation of randomized control trials results in this manner is that collection of accurate data on the number of discrete exposure events is not always possible and that there is substantial heterogeneity in degree of risk exposure between participants in trials. CONCLUSIONS: The implications of this analysis are that common interpretations of clinical trial interventions are insufficient for understanding the true efficacy of an intervention in some circumstances. Estimates of effectiveness in trials may misrepresent effectiveness per exposure event and effectiveness over a lifetime of risk. Care should be taken when designing protocols for analysis of trial results when the expected incidence is high. No change to the current practice of designing randomized control trials is suggested but analysis of trial data could be extended to calculate other statistics of effectiveness. A type of extrapolation and interpolation method for estimating levels of effectiveness is proposed. PMID- 20007406 TI - Pharmacogenomic approach reveals a role for the x(c)- cystine/glutamate antiporter in growth and celastrol resistance of glioma cell lines. AB - The x(c)(-) cystine/glutamate antiporter has been implicated in GSH-based chemoresistance because it mediates cellular uptake of cystine/cysteine for sustenance of intracellular GSH levels. Celastrol, isolated from a Chinese medicinal herb, is a novel heat shock protein 90 (Hsp90) inhibitor with potent anticancer activity against glioma in vitro and in vivo. In search of correlations between growth-inhibitory potency of celastrol in NCI-60 cell lines and microarray expression profiles of most known transporters, we found that expression of SLC7A11, the gene encoding the light chain subunit of x(c)(-), showed a strong negative correlation with celastrol activity. This novel gene drug correlation was validated. In celastrol-resistant glioma cells that highly expressed SLC7A11, sensitivity to celastrol was consistently increased via treatment with x(c)(-) inhibitors, including glutamate, (S)-4 carboxyphenylglycine, sulfasalazine, and SLC7A11 small interfering RNA. The GSH synthesis inhibitor, buthionine sulfoximine, also increased celastrol sensitivity, whereas the GSH booster, N-acetylcysteine, suppressed its cytotoxicity. Furthermore, the glioma cell lines were dependent on x(c)(-) mediated cystine uptake for viability, because cystine omission from the culture medium resulted in cell death and treatment with sulfasalazine depleted GSH levels and inhibited their growth. Combined treatment of glioma cells with sulfasalazine and celastrol led to chemosensitization, as suggested by increased celastrol-induced cell cycle arrest, apoptosis, and down-regulation of the Hsp90 client protein, epidermal growth factor receptor. These results indicate that the x(c)(-) transporter provides a useful target for glioma therapy. x(c)(-) inhibitors such as sulfasalazine, a Food and Drug Administration-approved drug, may be effective both as an anticancer drug and as an agent for sensitizing gliomas to celastrol. PMID- 20007408 TI - Redesigning a large-scale clinical trial in response to negative external trial results: the CAMUS study of phytotherapy for benign prostatic hyperplasia. AB - BACKGROUND: Benign prostatic hyperplasia (BPH), a common condition among older men, confers its morbidity through potentially bothersome lower urinary tract symptoms. Treatments for BPH include drugs such as alpha-adrenergic receptor blockers and 5-alpha reductase inhibitors, minimally invasive therapies that use heat to damage or destroy prostate tissue, and surgery including transurethral resection of the prostate. Complementary and alternative medicines are gaining popularity in the US. Two phytotherapies commonly used for BPH are extracts of the fruit of Serenoa repens, the Saw palmetto dwarf palm that grows in the Southeastern US, and extracts of the bark of Pygeum africanum, the African plum tree. PURPOSE: The objective of the Complementary and Alternative Medicines for Urological Symptoms (CAMUS) clinical trial is to determine if phytotherapy is superior to placebo in the treatment of BPH. METHODS: CAMUS was originally designed as a 3300-participant, four-arm trial of S. repens, P. africanum, an alpha-adrenergic blocking drug, and placebo with time to clinical progression of BPH, a measure of long-term efficacy, as the primary endpoint. Before enrollment started, a randomized, double-blind, placebo-controlled, single institution clinical trial showed that S. repens at the usual dose did not demonstrate any benefit over placebo with respect to symptom relief at 1 year. Consequently, the focus of CAMUS shifted from evaluating long-term efficacy to determining if any short-term (6-18 months) symptom relief could be achieved with increasing doses of S. repens, the phytotherapy most commonly used in the US for BPH. RESULTS: Results are anticipated in 2011. CONCLUSIONS: Trial design occurs in an environment of continually evolving information. In this case, emerging results from another trial suggested that a study of long-term efficacy was premature, and that an effective dose and preparation of S. repens had to be established before proceeding to a long-term clinical trial. PMID- 20007409 TI - Comparative contribution of the aryl hydrocarbon receptor gene to perinatal stage development and dioxin-induced toxicity between the urogenital complex and testis in the mouse. AB - TCDD (2,3,7,8-tetrachlorodebenzo-p-dioxin) requires the presence of the aryl hydrocarbon receptor (Ahr) gene for its toxic effects, such as reproductive disorders in male offspring of maternally exposed rats and mice. To study the involvement of the Ahr gene in producing the toxic phenotype with respect to testicular development, we administered a relatively high dose of TCDD to mice with three different maternally derived Ahr genotypic traits, and then compared several Ahr-dependent alterations among male reproductive systems on Postnatal Day 14. Reduction in anogenital distance and expression of prostatic epithelial genes in the urogenital complex (UGC) were detected in Ahr(+/+) and Ahr(+/-) mice exposed to TCDD, whereas no difference was observed in Ahr(-/-) mice. In situ hybridization revealed the absence of probasin mRNA expression in the prostate epithelium, despite the obvious development of prostatic lobes in TCDD-exposed mice. In contrast to obvious prostatic dysfunction and induction of cytochrome P450 (CYP) family genes in the UGC by TCDD, no alterations in testicular functions were observed in germ cell/Sertoli cell/interstitial cell marker gene expression or CYP family induction. No histopathological changes were observed among the three genotypes and between control and TCDD-exposed mice. Therefore, mouse external genitalia and prostatic development are much more sensitive to TCDD treatment than testis. Further, the Ahr gene, analyzed in this study, does not significantly contribute to testicular function during perinatal and immature stages, and the developing mouse testis appears to be quite resistant to TCDD exposure. PMID- 20007410 TI - Development and characterization of a three-dimensional organotypic human vaginal epithelial cell model. AB - We have developed an in vitro human vaginal epithelial cell (EC) model using the innovative rotating wall vessel (RWV) bioreactor technology that recapitulates in vivo structural and functional properties, including a stratified squamous epithelium with microvilli, tight junctions, microfolds, and mucus. This three dimensional (3-D) vaginal model provides a platform for high-throughput toxicity testing of candidate microbicides targeted to combat sexually transmitted infections, effectively complementing and extending existing testing systems such as surgical explants or animal models. Vaginal ECs were grown on porous, collagen coated microcarrier beads in a rotating, low fluid-shear environment; use of RWV bioreactor technology generated 3-D vaginal EC aggregates. Immunofluorescence and scanning and transmission electron microscopy confirmed differentiation and polarization of the 3-D EC aggregates among multiple cell layers and identified ultrastructural features important for nutrient absorption, cell-cell interactions, and pathogen defense. After treatment with a variety of toll-like receptor (TLR) agonists, cytokine production was quantified by cytometric bead array, confirming that TLRs 2, 3, 5, and 6 were expressed and functional. The 3-D vaginal aggregates were more resistant to nonoxynol-9 (N-9), a contraceptive and previous microbicide candidate, when compared to two-dimensional monolayers of the same cell line. A dose-dependent production of tumor necrosis factor-related apoptosis-inducing ligand and interleukin-1 receptor antagonist, biomarkers of cervicovaginal inflammation, correlated to microbicide toxicity in the 3-D model following N-9 treatment. These results indicate that this 3-D vaginal model could be used as a complementary tool for screening microbicide compounds for safety and efficacy, thus improving success in clinical trials. PMID- 20007411 TI - Increased expression of enzymes for sphingosine 1-phosphate turnover and signaling in human decidua during late pregnancy. AB - An appropriate balance between uterine quiescence and activation during pregnancy is essential for a successful outcome. Sphingosine 1-phosphate (S1P), a bioactive lipid, increases cell survival, proliferation, and angiogenesis, all important to maintain the pregnancy. Indeed progesterone increases sphingosine kinase 1 (SPHK1) mRNA, which produces S1P. In contrast, induction of prostaglandin endoperoxide synthase 2 by S1P and stimulation of SPHK1 by estradiol and cytokines suggests a role for S1P in the termination of pregnancy. Human decidua is important for regulating the maintenance and termination of pregnancy with production of progesterone receptors, cytokines, and prostaglandins. We hypothesized that S1P is produced by and acts on the decidua to stimulate production of mediators that induce labor. Our objective was to investigate the metabolism of S1P and its receptors in human decidua during pregnancy. We found that SPHK1 protein and activity positively correlated with increasing gestational age in human decidua parietalis. This was accompanied at term by increased expression of the S1P lyase, which irreversibly degrades S1P. This implies increased S1P turnover in the decidua at term. Although the mRNA level of phosphatidic acid phosphatase type 2A and 2B (PPAP2A,B), which dephosphorylate extracellular S1P, were increased at term, PPAP2 activity did not change. Sphingosine 1-phosphate receptor 3 protein expression also increased at term, indicating increased signaling by S1P in the decidua. There were no differences in any parameter tested in decidua from women in labor compared to those who were not. This work provides the first evidence of increased S1P synthesis, degradation, and signaling in human decidua during gestation. PMID- 20007412 TI - Why anguish? PMID- 20007413 TI - Neuropsychological consequences of alcohol and drug abuse on different components of executive functions. AB - Several studies have shown alterations in different components of executive functioning in users of different drugs, including cannabis, cocaine and heroin. However, it is difficult to establish a specific association between the use of each of these drugs and executive alterations, since most drug abusers are polysubstance abusers, and alcohol is a ubiquitous confounding factor. Moreover, in order to study the association between consumption of different drugs and executive functioning, the patterns of quantity and duration of drugs used must be considered, given the association between these parameters and the executive functioning alteration degree. Based on the multicomponent approach to executive functions, the aims of the present study were: (i) to analyse the differential contribution of alcohol versus cocaine, heroin and cannabis use on executive functions performance; and (ii) to analyse the contribution made by the severity of the different drugs used (quantity and duration patterns) on these functions in a sample of polysubstance abusers that requested treatment for cannabis-, cocaine- or heroin-related problems. We administered measures of fluency, working memory, analogical reasoning, interference, cognitive flexibility, decision making and self-regulation to two groups: 60 substance-dependent individuals (SDIs) and 30 healthy control individuals (HCIs). SDIs had significantly poorer performance than HCIs across all of the executive domains assessed. Results from hierarchical regression models showed the existence of common correlates of the use of alcohol, cannabis and cocaine on verbal fluency and decision-making; common correlates of quantity of cannabis and cocaine use on verbal working memory and analogical reasoning; common correlates of duration of cocaine and heroin use on shifting; and specific effects of duration of cocaine use on inhibition measures. These findings indicate that alcohol abuse is negatively associated with fluency and decision-making deficits, whereas the different drugs motivating treatment have both generalized and specific deleterious effects on different executive components. PMID- 20007414 TI - Madelung's deformity: a review. AB - Madelung's deformity is a rare condition of the wrist characterized by a shortened distal radius with volar-ulnar curvature and a dorsally prominent distal ulna. It occurs predominantly in adolescent females who present with pain, decreased wrist mobility, and deformity. Although its aetiology remains unclear, its treatment is becoming more refined. Several different surgical techniques have recently been described in the literature. This review addresses Madelung's deformity and suggests an algorithm for management based on current literature and the authors' own clinical experience. PMID- 20007415 TI - Warm or refrigerated local anaesthetic for open carpal tunnel release: a single blind randomized controlled study. AB - The aim of this randomized controlled study was to determine whether administration of lignocaine with adrenaline is less painful when injected at room temperature compared to refrigerated temperature. A cohort of 50 patients undergoing unilateral carpal tunnel decompression was randomized to room temperature or refrigerated local anaesthetic. Pain scores were assessed using a 10 mm visual analogue scale. Mean pain scores were 4.0 (SD +/- 1.5) for room temperature and 6.5 (SD +/- 1.7) for refrigerated local anaesthetic (P < 0.001). This study demonstrates that patients experience greater pain levels with administration of local anaesthetic at refrigerated temperatures prior to open carpal tunnel release. PMID- 20007416 TI - The extensor tendon splitting approach to the proximal interphalangeal joint: do we need to reinsert the central slip? AB - Current approaches to the proximal interphalangeal (PIP) joint have potential complications and limitations. We present a dorsal approach that involves splitting the extensor tendon in the midline, detaching the insertion of the central slip and repairing the extensor tendon without reinserting the tendon into the base of the middle phalanx. A retrospective review of 16 digits that had the approach for a PIP joint arthroplasty with a mean follow up of 23 months found a postoperative PIP active ROM of 61 degrees (range 25-90 degrees). Fourteen digits had no extensor lag, while two digits had an extensor lag of 20 degrees and 25 degrees. This modified approach is fast and simple and does not cause an extensor lag. PMID- 20007418 TI - Co-existing carpal tunnel syndrome in complex regional pain syndrome after hand trauma. AB - This study highlights the benefits of carpal tunnel release (CTR) in four patients presenting with complex regional pain syndrome (CRPS) after hand surgery who also had carpal tunnel syndrome (CTS) diagnosed clinically and by nerve conduction studies. Three of the patients underwent pre- and postoperative volumetric, circumference, grip strength and range of motion measurements. The Disabilities of the Arm, Shoulder and Hand (DASH) functional outcome measure and pain scores were also used. There was almost complete resolution of CRPS symptoms in all four patients, with notable reductions in oedema and improvements in grip strength and range of motion. There were also improvements in DASH outcome scores and pain scores after CTR. PMID- 20007419 TI - Anomalous presentation of flexor carpi radialis brevis: a report of six cases. AB - An anomalous presentation of flexor carpi radialis brevis (FCRB) is reported in six patients. These findings occurred in a consecutive series of 172 distal radius fracture fixations using a volar approach between 2002 and 2007. This is the first report of this anomalous muscle from a clinical series. PMID- 20007420 TI - The association between intraoperative correction of Dupuytren's disease and residual postoperative contracture. AB - The purpose of this study was to determine whether preoperative contracture and the amount of intraoperative correction can be used to predict the postoperative outcome of fasciectomy for Dupuytren's disease. A prospective study of 52 patients undergoing primary fasciectomy during an 18 month period was undertaken. The contracture of each joint was measured preoperatively, after fasciectomy during the operation and 6 months after surgery. Forty-two metacarpophalangeal (MCPJ) and 58 proximal interphalangeal (PIPJ) joints were treated surgically. Full intraoperative correction was achieved in 41 MCPJs. Thirty-seven had full correction at follow-up. Full intraoperative correction was obtained in 35 PIPJs and 13 had complete correction at follow-up. The extent of the preoperative deformity was a significant predictor of complete intraoperative correction. The extent of both preoperative deformity and intraoperative correction were significant predictors of loss of surgical correction after operation. PMID- 20007421 TI - The distribution of the types of thumb polydactyly in a Middle Eastern population: a study of 228 hands. AB - Several series have reported the distribution of the types of thumb polydactyly in the Caucasian and Far Eastern populations. No data are available for the Middle East. A total of 196 Saudi patients (228 hands) with thumb polydactyly were reviewed. The most common type was Wassel type IV (33.8%) and the least common was Wassel type I (0.4%). A total of 26 hands (11.4%) did not fit into the classic Wassel types including 18 cases of rudimentary duplications, two cases of thumb triplication, and five cases of thumb duplication with symphalangism. Concurrent ipsilateral little finger triplication (one case) and contralateral thumb aplasia/hypoplasia (two cases) were seen and the developmental biology of these cases are discussed. After reviewing our results and other series in the literature, we concluded that Wassel types IV and I remain to be the most and least common types respectively in almost all races. However, the genetic pools of various races greatly affect the distribution of other non-classic duplications (unclassifiable by Wassel classification) such as rudimentary duplication in the Far East, triplication in the Southwestern region of the Netherlands, and symphalangism in Asia. PMID- 20007422 TI - Use of human amniotic membrane wrap in reducing perineural adhesions in a rabbit model of ulnar nerve neurorrhaphy. AB - The object of this experimental study was to assess the effect of wrapping human amniotic membrane around a repaired ulnar nerve in a rabbit model of perineural adhesion. Ulnar nerves from 10 white New Zealand rabbits were exposed bilaterally, dissected and repaired. Human amniotic membrane was then wrapped around the repair site in one limb with no such wrap in the neurorrhaphy of the contralateral limb. Three months later, the same nerves were re-explored and removed using microsurgical external neurolysis. Perineural adhesion around the ulnar nerve was evaluated by blinded surgical dissection and scored using a visual 4-point qualitative scale. Extent and grade of fibrosis around repair sites were measured microscopically (x 200) after Masson trichrome staining using measure of the depth of fibrosis and the grading criteria of adhesion. Quantitative morphometric analysis was also performed under light microscopy (x 200) with the aid of a digital counter and virtual slide imaging software (ScanScope T2, Vista, CA, USA). Human amniotic membrane wrapped nerves showed significantly less perineural adhesion and fibrosis than controls (P < 0.05). No nerve healing problems were encountered. This study suggests that human amniotic membrane application can reduce fibrosis and adhesion around neurorrhaphy sites in this animal model. PMID- 20007423 TI - Ulnar variance and scaphoid fracture. AB - Between 1997 and 2006, radiographs of 66 scaphoid fractures were retrospectively reviewed to evaluate ulnar variance. Twenty-one (31.8%) patients had an 'ulna neutral' wrist, six (9.1%) had an 'ulna plus' and 39 (59.1%) had an 'ulna minus' wrist. The mean ulnar variance was -1.3 (SD 1.8) mm (range -5.5, 2.5). We observed a significant difference in the distribution of ulnar variance (P < 0.00001) and in the proportion of cases with ulna minus (OR = 5.0; 95% CI: 2.7, 9.3) compared to previous publications. PMID- 20007424 TI - A minimally invasive anterior approach to reduction and screw fixation of coronoid fractures. AB - We investigated a minimally invasive anterior approach to reduce and fix coronoid fractures with a screw in eight consecutive patients and evaluated seven patients at a minimum of 1 year (mean, 78 weeks; range, 61-89 weeks). It was only possible to repair seven of eight coronoid fractures. The average time to radiographic healing was 12 weeks and all healed without the screw loosening, migration or breakage. Endoscopy permitted an excellent view of coronoid fractures, allowing anatomic fixation, and as it avoided extensive soft tissue dissection, this method preserved soft tissue attachments of small coronoid fragments. This method was safe and minimally invasive. PMID- 20007425 TI - Diffusion tensor imaging abnormalities in depressed multiple sclerosis patients. AB - Depression is common in patients with multiple sclerosis, but to date no studies have explored diffusion tensor imaging indices associated with mood change. This study aimed to determine cerebral correlates of depression in multiple sclerosis patients using diffusion tensor imaging. Sixty-two subjects with multiple sclerosis were assessed for depression with the Beck Depression Inventory (BDI II). All subjects underwent magnetic resonance imaging. Whole brain and regional volumes were calculated for lesions (hyper/hypointense) and normal-appearing white and grey matter. Fractional anisotropy and mean diffusivity were calculated for each brain region. Magnetic resonance imaging comparisons were undertaken between depressed (Beck Depression Inventory > or = 19) and non-depressed subjects. Depressed subjects (n = 30) had a higher hypointense lesion volume in the right medial inferior frontal region, a smaller normal-appearing white matter volume in the left superior frontal region, and lower fractional anisotropy and higher mean diffusivity in the left anterior temporal normal-appearing white matter and normal-appearing grey matter regions, respectively. Depressed subjects also had higher mean diffusivity in right inferior frontal hyperintense lesions. Magnetic resonance imaging variables contributed to 43% of the depression variance. We conclude that the presence of more marked diffusion tensor imaging abnormalities in the normal-appearing white matter and normal-appearing grey matter of depressed subjects highlights the importance of more subtle measures of structural brain change in the pathogenesis of depression. PMID- 20007426 TI - The cannabinoid receptor 1 gene (CNR1) and multiple sclerosis: an association study in two case-control groups from Spain. AB - Different studies point to the implication of the endocannabinoid system in multiple sclerosis (MS) and animal models of MS. The purpose of this study was to evaluate a possible association of MS with polymorphic markers at the CNR1 gene, encoding the cannabinoid 1 (CB(1)) receptor. We have performed a genetic analysis of an AAT repeat microsatellite localized in the downstream region of the CNR1 gene, in two case-control groups of MS patients and healthy controls (HC) from Spain (Madrid and Bilbao). MS patients with primary progressive MS (PPMS) had more commonly long ((AAT) > or = (13)) alleles and genotypes with a significant difference for genotype 7/8 in Madrid (p = 0.043) and in the sum of both groups (p = 0.016); short alleles were less frequently found in PPMS with a significant difference for allele 5 in the analysis of both groups together (p = 0.039). In patients with relapsing MS, no consistent differences in allele and genotype distribution were found. Disease severity and progression was unrelated to AAT repeat variations. In conclusion, long (AAT) > or = (13) CNR1 genotypes could behave as risk factors for PPMS. PMID- 20007427 TI - Immunoglobulin-like transcript 3, an inhibitor of T cell activation, is reduced on blood monocytes during multiple sclerosis relapses and is induced by interferon beta-1b. AB - Immunoglobulin-like transcripts (ILTs) are immunoregulatory proteins that either activate or inhibit immune responses. ILT3 is inhibitory and is expressed preferentially by antigen-presenting cells. When its extracellular domain binds to an unidentified ligand of activated T cells, the T cell is silenced. Our objective was to study the expression of ILT3 on circulating monocytes in RRMS. Freshly isolated peripheral blood mononuclear cells were analyzed by multicolored flow cytometry. The proportion of ILT3(+)CD14(+) monocytes in blood, and ILT3 levels expressed by them, is lower in untreated multiple sclerosis in relapse than in: (1) untreated multiple sclerosis in remission (p < 0.009); (2) stable interferon beta-treated relapsing-remitting multiple sclerosis (p < 0.001) and; (3) healthy controls (p < 0.009). Glatiramer acetate-stimulated CD4( +) T cells, co-cultured with freshly isolated monocytes, proliferate significantly better (p = 0.0017 for multiple sclerosis; p = 0.0015 for controls) when T cell interaction with monocyte-expressed ILT3 is blocked by anti-ILT3 antibody. Interferon beta is beneficial in multiple sclerosis; why so remains unclear. Interferon beta-1b markedly increases ILT3 expression in vitro by monocytes from multiple sclerosis patients and controls. These findings identify a putative novel mechanism for the therapeutic benefit bestowed by Interferon beta and a new target for therapeutic intervention in relapsing-remitting multiple sclerosis. PMID- 20007428 TI - Renewal of the T-cell compartment in multiple sclerosis patients treated with glatiramer acetate. AB - The immunomodulating activity of glatiramer acetate on T-cells of multiple sclerosis patients has only been partially clarified. The objective of this work was to investigate whether glatiramer acetate modifies thymic release of newly produced T-cells and the peripheral composition of the T-cell repertoire. T-cell receptor excision circles, (thymic) naive (CD4(+)CD45RA(+)CCR7(+)CD31(+)) T helper cells, and central (CD4(+)CD45RA(-)CCR7(+)) and effector (CD4(+)CD45RA( )CCR7(-)) memory T-cells were evaluated in 89 untreated patients, 84 patients treated for at least 1 year, and 31 patients beginning treatment at the time of inclusion in the study and then followed-up for 12 months; controls were 81 healthy donors. The T-cell repertoire was analysed in selected samples. The percentage of (thymic)naive T helper cells was diminished in untreated patients, but rose to control values in treated subjects; a decrease in central memory T cells was also observed in treated patients. Follow-up patients could be divided into two subgroups, one showing unmodified (thymic)naive T helper cells and T cell diversity, the other in which the increased release of new T-cells was accompanied by modifications of the T-cell repertoire. Glatiramer acetate modifies the peripheral T-cell pool by activating a thymopoietic pathway of T cell release that leads to a different setting of T-cell diversity and, likely, to a dilution of autoreactive T-cells. PMID- 20007429 TI - Multiparametric MR investigation of the motor pyramidal system in patients with 'truly benign' multiple sclerosis. AB - One possible explanation for the mismatch between tissue damage and preservation of neurological functions in patients with benign multiple sclerosis (BMS) is that the pathophysiology differs from that occurring in other multiple sclerosis (MS) phenotypes. The objective of this study was to identify pathologically specific patterns of tissue integrity/damage characteristics of patients with BMS, and markers of potential prognostic value. The pyramidal system was investigated in 10 BMS patients and 20 controls using voxel-based morphometry to assess grey matter (GM) atrophy, and diffusion tractography and quantitative magnetization transfer to quantify the microstructural damage in the corticospinal tracts (CSTs). Widespread reductions in GM volume were found in patients compared with controls, including the primary motor cortex. A significant decrease was observed in the mean macromolecular pool ratio (F) of both CSTs, with no fractional anisotropy (FA) change. GM volume of the primary motor areas was associated with clinical scores but not with the CST parameters. The mismatch between F and FA suggests the presence of extensive demyelination in the CSTs of patients with BMS, in the absence of axonal damage. The lack of correlation with GM volume indicates a complex interaction between disruptive and reparative mechanisms in BMS. PMID- 20007430 TI - Social consequences of multiple sclerosis (1): early pension and temporary unemployment--a historical prospective cohort study. AB - Multiple sclerosis affects young and middle-aged people and often leads to physical and cognitive handicaps. There is a need for detailed knowledge of the social consequences of the disease. We aim here to describe the course of the working life and career of multiple sclerosis patients at the time of onset and thereafter, in terms of probability of early pension and income development. All 2538 patients with multiple sclerosis in Denmark with disease onset between 1980 and 1989, identified through the Danish MS-Registry, were included in this study. Twenty matched control persons per patient were randomly drawn from the civil registration system. Information on economic status was retrieved from Statistics Denmark. A survival analysis technique was used with onset as the starting point. We found that the probability of remaining without early pension was at 5 years 70% for patients and 97% for controls, and at 20 years 22% for patients and 86% for controls. Due to lower rates for early pension, gross income with time was lower in patients than controls. We conclude that multiple sclerosis seriously affects the economic life of multiple sclerosis patients, even within a few years of onset. PMID- 20007431 TI - Natalizumab treatment in multiple sclerosis: marked decline of chemokines and cytokines in cerebrospinal fluid. AB - Natalizumab exerts impressive therapeutic effects in patients with multiple sclerosis (MS). The proposed main mode of action is reducing transmigration of leukocytes into the CNS, but other immunological effects may also be operative. Cytokines and chemokines are involved in the regulation of inflammatory responses and may reflect the disease process in MS. The objective of this study was to evaluate the effects of natalizumab treatment on cytokine and chemokine profiles systemically and intrathecally in multiple sclerosis. We used luminex to analyse a panel of cytokines (IL-1beta, IL-2, IL-4, IL-5, IL-6, IL-8, IL-10, TNF-alpha, IFN-gamma, GM-CSF) and chemokines (CXCL9, CXCL10, CXCL11, CCL17, CCL22) in blood and cerebrospinal fluid (CSF) from 31 patients with relapsing MS before and after one year of natalizumab treatment. There was a marked decline in CSF levels of cytokines and chemokines, thus including pro-inflammatory cytokines (IL-1beta, IL 6 and IL-8) as well as chemokines associated with both Th1 (CXCL9, CXCL10, CXCL11) and Th2 (CCL22). Circulating plasma levels of some cytokines (GM-CSF, TNF alpha, IL-6 and IL-10) also decreased after one year of treatment. This is the first study to show that natalizumab treatment is associated with a global decline in cytokine and chemokine levels at a protein level. This finding was most pronounced in CSF, in line with the reduced transmigration of cells into CNS, whereas reduction in plasma levels indicates other possible mechanisms of natalizumab treatment. PMID- 20007432 TI - Polymorphisms in vitamin D metabolism related genes and risk of multiple sclerosis. AB - The extent to which potential genetic determinants of vitamin D levels may be related to multiple sclerosis (MS) risk has not been thoroughly explored. The objective of this study was to determine whether polymorphisms in VDR, CYP27B1, CYP24A1, CYP2R1 and DBP are associated with the risk of MS and whether these variants may modify associations between environmental or dietary vitamin D on MS risk. A nested case-control study was conducted in two, large cohorts of US nurses, including 214 MS cases and 428 age-matched controls. Conditional logistic regression models were used to calculate relative risks (RR) and 95% confidence intervals (CIs) and to assess the significance of gene-environment interactions. No associations were observed for any of the single-nucleotide polymorphisms (SNPs) in VDR, CYP27B1, CYP24A1, CYP2R1 or DBP (p > 0.05 for all). The authors did observe an interaction (p = 0.04) between dietary intake of vitamin D and the vitamin D receptor FokI polymorphism on MS risk. The protective effect of increasing vitamin D was evident only in individuals with the 'ff ' genotype (RR = 0.2, 95% CI: 0.06, 0.78; p = 0.02 for 400 IU/day increase). It was concluded that this does not support a role for the selected SNPs involved in vitamin D metabolism in the etiology of MS. The finding of a marginally significant gene environment interaction requires replication in larger datasets, but suggests future genetic studies may benefit from considering relevant environmental context. PMID- 20007433 TI - 'Gimme five': future challenges in multiple sclerosis. ECTRIMS Lecture 2009. AB - This article is based on the ECTRIMS lecture given at the 25( th) ECTRIMS meeting which was held in Dusseldorf, Germany, from 9 to 12 September 2009. Five challenges have been identified: (1) safeguarding the principles of medical ethics; (2) optimizing the risk/benefit ratio; (3) bridging the gap between multiple sclerosis and experimental autoimmune encephalitis; (4) promoting neuroprotection and repair; and (5) tailoring multiple sclerosis therapy to the individual patient. Each of these challenges will be discussed and placed in the context of current research into the pathogenesis and treatment of multiple sclerosis. PMID- 20007440 TI - Oscillatory growth in lily pollen tubes does not require aerobic energy metabolism. AB - Oscillatory tip growth in pollen tubes depends on prodigious amounts of energy. We have tested the hypothesis that oscillations in the electron transport chain lead to growth oscillations in lily (Lilium formosanum). Using three respiratory inhibitors, oligomycin, antimycin A, and cyanide, we find that pollen tube growth is much less sensitive to respiratory inhibition than respiration is. All three block respiration at concentrations severalfold lower than necessary to inhibit growth. Mitochondrial NAD(P)H and potentiometric JC-1 fluorescence, employed as markers for electron transport chain activity, rise rapidly in response to oligomycin, as expected. Pollen tube growth stops for several minutes before resuming. Subsequent growth has a lower mean rate, but continues to oscillate, albeit with a longer period. NAD(P)H fluorescence no longer exhibits coherent oscillations, and mitochondria no longer congregate directly behind the apex: they distribute evenly throughout the cell. Postinhibition growth relies on aerobic fermentation for energy production as revealed by an increase in ethanol in the media. These data suggest that oscillatory growth depends not on a single oscillatory pacemaker but rather is an emergent property arising from a number of stable limit cycles. PMID- 20007441 TI - Systems biology of organ initiation at the shoot apex. PMID- 20007442 TI - Phosphoenolpyruvate carboxylase from C4 leaves is selectively targeted for inhibition by anionic phospholipids. PMID- 20007443 TI - Knockdown of the symbiotic sucrose synthase MtSucS1 affects arbuscule maturation and maintenance in mycorrhizal roots of Medicago truncatula. AB - The relevance of the symbiosis-induced Medicago truncatula sucrose synthase gene MtSucS1 for an efficient arbuscular mycorrhiza (AM) was studied using two independent antisense lines that displayed up to 10-fold reduced SucS1 levels in roots. Mycorrhizal MtSucS1-reduced lines exhibited an overall stunted aboveground growth under inorganic phosphorus limitation. Apart from a reduced plant height, shoot weight, and leaf development, a delayed flowering, resulting in a lower seed yield, was observed. In addition, the root-to-shoot and root weight ratios increased significantly. Gene expression studies demonstrated a major reversion of AM-associated transcription, exhibiting a significant repression of well-known plant AM marker and mycosymbiont genes, together indicating a diminished AM fungus colonization of MtSucS1-antisense lines. Concomitantly, gas chromatography mass spectrometry-based metabolite profiling revealed that mycorrhizal MtSucS1 reduced lines were affected in important nodes of the carbon, nitrogen, and phosphorus metabolism, accentuating a physiological significance of MtSucS1 for AM. In fact, antisensing MtSucS1 provoked an impaired fungal colonization within the less abundant infected regions, evident from strongly reduced frequencies of internal hyphae, vesicles, and arbuscules. Moreover, arbuscules were early senescing, accompanied with a reduced development of mature arbuscules. This defective mycorrhiza status correlated with reduced phosphorus and nitrogen levels and was proportional to the extent of MtSucS1 knockdown. Together, our results point to an important role for MtSucS1 in the establishment and maintenance of arbuscules in the AM symbiosis. PMID- 20007444 TI - A Raf-like MAPKKK gene DSM1 mediates drought resistance through reactive oxygen species scavenging in rice. AB - Mitogen-activated protein kinase (MAPK) cascades have been identified in various signaling pathways involved in plant development and stress responses. We identified a drought-hypersensitive mutant (drought-hypersensitive mutant1 [dsm1]) of a putative MAPK kinase kinase (MAPKKK) gene in rice (Oryza sativa). Two allelic dsm1 mutants were more sensitive than wild-type plants to drought stress at both seedling and panicle development stages. The dsm1 mutants lost water more rapidly than wild-type plants under drought stress, which was in agreement with the increased drought-sensitivity phenotype of the mutant plants. DSM1-RNA interference lines were also hypersensitive to drought stress. The predicted DSM1 protein belongs to a B3 subgroup of plant Raf-like MAPKKKs and was localized in the nucleus. By real-time PCR analysis, the DSM1 gene was induced by salt, drought, and abscisic acid, but not by cold. Microarray analysis revealed that two peroxidase (POX) genes, POX22.3 and POX8.1, were sharply down-regulated compared to wild type, suggesting that DSM1 may be involved in reactive oxygen species (ROS) signaling. Peroxidase activity, electrolyte leakage, chlorophyll content, and 3,3'-diaminobenzidine staining revealed that the dsm1 mutant was more sensitive to oxidative stress due to an increase in ROS damage caused by the reduced POX activity. Overexpression of DSM1 in rice increased the tolerance to dehydration stress at the seedling stage. Together, these results suggest that DSM1 might be a novel MAPKKK functioning as an early signaling component in regulating responses to drought stress by regulating scavenging of ROS in rice. PMID- 20007445 TI - Chitinase-like protein CTL1 plays a role in altering root system architecture in response to multiple environmental conditions. AB - Plant root architecture is highly responsive to changes in nutrient availability. However, the molecular mechanisms governing the adaptability of root systems to changing environmental conditions is poorly understood. A screen for abnormal root architecture responses to high nitrate in the growth medium was carried out for a population of ethyl methanesulfonate-mutagenized Arabidopsis (Arabidopsis thaliana). The growth and root architecture of the arm (for anion altered root morphology) mutant described here was similar to wild-type plants when grown on low to moderate nitrate concentrations, but on high nitrate, arm exhibited reduced primary root elongation, radial swelling, increased numbers of lateral roots, and increased root hair density when compared to the wild-type control. High concentrations of chloride and sucrose induced the same phenotype. In contrast, hypocotyl elongation in the dark was decreased independently of nitrate availability. Positional cloning identified a point mutation in the AtCTL1 gene that encodes a chitinase-related protein, although molecular and biochemical analysis showed that this protein does not possess chitinase enzymatic activity. CTL1 appears to play two roles in plant growth and development based on the constitutive effect of the arm mutation on primary root growth and its conditional impact on root architecture. We hypothesize that CTL1 plays a role in determining cell wall rigidity and that the activity is differentially regulated by pathways that are triggered by environmental conditions. Moreover, we show that mutants of some subunits of the cellulose synthase complex phenocopy the conditional effect on root architecture under nonpermissive conditions, suggesting they are also differentially regulated in response to a changing environment. PMID- 20007446 TI - Jasmonate-dependent and -independent pathways mediate specific effects of solar ultraviolet B radiation on leaf phenolics and antiherbivore defense. AB - Ultraviolet B (UV-B) radiation, a very small fraction of the daylight spectrum, elicits changes in plant secondary metabolism that have large effects on plant insect interactions. The signal transduction pathways that mediate these specific effects of solar UV-B are not known. We examined the role of jasmonate signaling by measuring responses to UV-B in wild-type and transgenic jasmonate-deficient Nicotiana attenuata plants in which a lipoxygenase gene (NaLOX3) was silenced (as lox). In wild-type plants, UV-B failed to elicit the accumulation of jasmonic acid (JA) or the bioactive JA-isoleucine conjugate but amplified the response of jasmonate-inducible genes, such as trypsin proteinase inhibitor (TPI), to wounding and methyl jasmonate, and increased the accumulation of several phenylpropanoid derivatives. Some of these phenolic responses (accumulation of caffeoyl-polyamine conjugates) were completely lacking in as-lox plants, whereas others (accumulation of rutin and chlorogenic acid) were similar in both genotypes. In open field conditions, as-lox plants received more insect damage than wild-type plants, as expected, but the dramatic increase in resistance to herbivory elicited by UV-B exposure, which was highly significant in wild-type plants, did not occur in as-lox plants. We conclude that solar UV-B (1) uses jasmonate-dependent and -independent pathways in the elicitation of phenolic compounds, and (2) increases sensitivity to jasmonates, leading to enhanced expression of wound-response genes (TPI). The lack of UV-B-induced antiherbivore protection in as-lox plants suggests that jasmonate signaling plays a central role in the mechanisms by which solar UV-B increases resistance to insect herbivores in the field. PMID- 20007447 TI - The role of casein kinase II in flowering time regulation has diversified during evolution. AB - Casein kinase II (CK2) is a protein kinase with an evolutionarily conserved function as a circadian clock component in several organisms, including the long day plant Arabidopsis (Arabidopsis thaliana). The circadian clock component CIRCADIAN CLOCK ASSOCIATED1 (CCA1) is a CK2 target in Arabidopsis, where it influences photoperiodic flowering. In rice (Oryza sativa), a short-day plant, Heading date6 (Hd6) encodes a CK2alpha subunit that delays flowering time under long-day conditions. Here, we demonstrate that control of flowering time in rice by the Hd6 CK2alpha subunit requires a functional Hd1 gene (an Arabidopsis CONSTANS ortholog) and is independent of the circadian clock mechanism. Our findings from overexpressing the dominant-negative CK2 allele in rice support the independence of CK2 function from the circadian clock. This lack of control of the circadian clock by Hd6 CK2alpha might be due to the presence of glutamate in OsLHY (a CCA1 ortholog in rice) instead of the serine at the corresponding CK2 target site in CCA1. However, this glutamate is critical for the control of the OsPRR1 gene (a rice ortholog of the Arabidopsis TOC1/PRR1 gene) by OsLHY for regulation of the circadian clock. We also demonstrated that the other conserved CK2 target sites in OsLHY conferred robust rhythmic expression of OsLHY-LUC under diurnal conditions. These findings imply that the role of CK2 in flowering-time regulation in higher plants has diversified during evolution. PMID- 20007448 TI - Enhanced abscisic acid-mediated responses in nia1nia2noa1-2 triple mutant impaired in NIA/NR- and AtNOA1-dependent nitric oxide biosynthesis in Arabidopsis. AB - Nitric oxide (NO) regulates a wide range of plant processes from development to environmental adaptation. Despite its reported regulatory functions, it remains unclear how NO is synthesized in plants. We have generated a triple nia1nia2noa1 2 mutant that is impaired in nitrate reductase (NIA/NR)- and Nitric Oxide Associated1 (AtNOA1)-mediated NO biosynthetic pathways. NO content in roots of nia1nia2 and noa1-2 plants was lower than in wild-type plants and below the detection limit in nia1nia2noa1-2 plants. NIA/NR- and AtNOA1-mediated biosynthesis of NO were thus active and responsible for most of the NO production in Arabidopsis (Arabidopsis thaliana). The nia1nia2noa1-2 plants displayed reduced size, fertility, and seed germination potential but increased dormancy and resistance to water deficit. The increasing deficiency in NO of nia1nia2, noa1-2, and nia1nia2noa1-2 plants correlated with increased seed dormancy, hypersensitivity to abscisic acid (ABA) in seed germination and establishment, as well as dehydration resistance. In nia1nia2noa1-2 plants, enhanced drought tolerance was due to a very efficient stomata closure and inhibition of opening by ABA, thus uncoupling NO from ABA-triggered responses in NO-deficient guard cells. The NO-deficient mutants in NIA/NR- and AtNOA1-mediated pathways in combination with the triple mutant will be useful tools to functionally characterize the role of NO and the contribution of both biosynthetic pathways in regulating plant development and defense. PMID- 20007449 TI - VirtualPlant: a software platform to support systems biology research. AB - Data generation is no longer the limiting factor in advancing biological research. In addition, data integration, analysis, and interpretation have become key bottlenecks and challenges that biologists conducting genomic research face daily. To enable biologists to derive testable hypotheses from the increasing amount of genomic data, we have developed the VirtualPlant software platform. VirtualPlant enables scientists to visualize, integrate, and analyze genomic data from a systems biology perspective. VirtualPlant integrates genome-wide data concerning the known and predicted relationships among genes, proteins, and molecules, as well as genome-scale experimental measurements. VirtualPlant also provides visualization techniques that render multivariate information in visual formats that facilitate the extraction of biological concepts. Importantly, VirtualPlant helps biologists who are not trained in computer science to mine lists of genes, microarray experiments, and gene networks to address questions in plant biology, such as: What are the molecular mechanisms by which internal or external perturbations affect processes controlling growth and development? We illustrate the use of VirtualPlant with three case studies, ranging from querying a gene of interest to the identification of gene networks and regulatory hubs that control seed development. Whereas the VirtualPlant software was developed to mine Arabidopsis (Arabidopsis thaliana) genomic data, its data structures, algorithms, and visualization tools are designed in a species-independent way. VirtualPlant is freely available at www.virtualplant.org. PMID- 20007450 TI - WallGen, software to construct layered cellulose-hemicellulose networks and predict their small deformation mechanics. AB - We understand few details about how the arrangement and interactions of cell wall polymers produce the mechanical properties of primary cell walls. Consequently, we cannot quantitatively assess if proposed wall structures are mechanically reasonable or assess the effectiveness of proposed mechanisms to change mechanical properties. As a step to remedying this, we developed WallGen, a Fortran program (available on request) building virtual cellulose-hemicellulose networks by stochastic self-assembly whose mechanical properties can be predicted by finite element analysis. The thousands of mechanical elements in the virtual wall are intended to have one-to-one spatial and mechanical correspondence with their real wall counterparts of cellulose microfibrils and hemicellulose chains. User-defined inputs set the properties of the two polymer types (elastic moduli, dimensions of microfibrils and hemicellulose chains, hemicellulose molecular weight) and their population properties (microfibril alignment and volume fraction, polymer weight percentages in the network). This allows exploration of the mechanical consequences of variations in nanostructure that might occur in vivo and provides estimates of how uncertainties regarding certain inputs will affect WallGen's mechanical predictions. We summarize WallGen's operation and the choice of values for user-defined inputs and show that predicted values for the elastic moduli of multinet walls subject to small displacements overlap measured values. "Design of experiment" methods provide systematic exploration of how changed input values affect mechanical properties and suggest that changing microfibril orientation and/or the number of hemicellulose cross-bridges could change wall mechanical anisotropy. PMID- 20007451 TI - SEUSS and SEUSS-LIKE transcriptional adaptors regulate floral and embryonic development in Arabidopsis. AB - Multimeric protein complexes are required during development to regulate transcription and orchestrate cellular proliferation and differentiation. The Arabidopsis (Arabidopsis thaliana) SEUSS (SEU) gene encodes a transcriptional adaptor that shares sequence similarity with metazoan Lim domain-binding transcriptional adaptors. In Arabidopsis, SEU forms a physical complex with the LEUNIG transcriptional coregulator. This complex regulates a number of diverse developmental events, including proper specification of floral organ identity and number and the development of female reproductive tissues derived from the carpel margin meristem. In addition to SEU, there are three Arabidopsis SEUSS-LIKE (SLK) genes that encode putative transcriptional adaptors. To determine the functions of the SLK genes and to investigate the degree of functional redundancy between SEU and SLK genes, we characterized available slk mutant lines in Arabidopsis. Here, we show that mutations in any single SLK gene failed to condition an obvious morphological abnormality. However, by generating higher order mutant plants, we uncovered a degree of redundancy between the SLK genes and between SLK genes and SEU. We report a novel role for SEU and the SLK genes during embryonic development and show that the concomitant loss of both SEU and SLK2 activities conditions severe embryonic and seedling defects characterized by a loss of the shoot apical meristem. Furthermore, we demonstrate that SLK gene function is required for proper development of vital female reproductive tissues derived from the carpel margin. We propose a model that posits that SEU and SLK genes support organ development from meristematic regions through two different pathways: one that facilitates auxin response and thus organ initiation and a second that sustains meristematic potential through the maintenance of SHOOTMERISTEM-LESS and PHABULOSA expression. PMID- 20007452 TI - Endothelial cells isolated from caveolin-2 knockout mice display higher proliferation rate and cell cycle progression relative to their wild-type counterparts. AB - The goal of this study was to determine whether caveolin-2 (Cav-2) is capable of controlling endothelial cell (EC) proliferation in vitro. To realize this goal, we have directly compared proliferation rates and cell cycle-associated signaling proteins between lung ECs isolated from wild-type (WT) and Cav-2 knockout (KO) mice. Using three independent proliferation assays, we have determined that Cav-2 KO ECs proliferate by ca. 2-fold faster than their WT counterparts. Cell cycle analysis by flow cytometry of propidium iodide-stained cells showed a relatively higher percentage of Cav-2 KO ECs in S and G(2)/M and lower percentage in G(o)/G(1) phases of cell cycle relative to their WT counterparts. Furthermore, an over 2-fold increase in the percentage of S phase-associated Cav-2 KO relative to WT ECs was independently determined with bromodeoxyuridine incorporation assay. Mechanistically, the increase in proliferation/cell cycle progression of Cav-2 KO ECs correlated well with elevated expression levels of predominantly S phase- and G(2)/M phase-associated cyclin A and B1, respectively. Further mechanistic analysis of molecular events controlling cell cycle progression revealed increased level of hyperphosphorylated (inactive) form of G(1) to S phase transition inhibitor, the retinoblastoma protein in hyperproliferating Cav-2 KO ECs. Conversely, the expression level of the two cyclin-dependent kinase inhibitors p16(INK4) and p27(Kip1) was reduced in Cav-2 KO ECs. Finally, increased phosphorylation (activation) of proproliferative extracellular signal regulated kinase 1/2 was observed in hyperproliferating Cav-2 KO ECs. Overall, our data suggest that Cav-2 negatively regulates lung EC proliferation and cell cycle progression. PMID- 20007453 TI - Tumor necrosis factor induces matrix metalloproteinases in cardiomyocytes and cardiofibroblasts differentially via superoxide production in a PI3Kgamma dependent manner. AB - Tumor necrosis factor (TNF) is an inflammatory cytokine that is upregulated in a number of cardiomyopathies. Adverse cardiac remodeling and dilation result from degradation of the extracellular matrix by matrix metalloproteinases (MMPs). We investigated whether TNF can directly trigger expression and activation of MMPs in cardiac cells. We compared MMP expression profile and activities between primary cultures of mouse neonatal cardiomyocytes and cardiofibroblasts and in cellular and extracellular compartments. In response to recombinant TNF (rTNF, 20 ng/ml), cardiomyocytes exhibited faster and more pronounced superoxide production compared with cardiofibroblasts, concomitant with increased expression of several MMPs. MMP9 levels increased more rapidly and about twofold more in cardiomyocytes than in cardiofibroblasts. TNF did not induce MMP2 expression. Expression of collagenases (MMP8, MMP12, MMP13, and MMP14) increased significantly, while total collagenase activity increased to a greater degree in conditioned medium of cardiomyocytes than in cardiofibroblasts. rTNF-mediated MMP expression and activation were dependent on superoxide production and were blocked by apocynin, an NADPH oxidase inhibitor. We identified phosphatidylinositol 3-kinase (PI3K)gamma as a key factor in TNF-mediated events since TNF-induced superoxide production, MMP expression, and activity were significantly suppressed in cardiomyocytes and cardiofibroblasts deficient in PI3Kgamma. We further demonstrated that the TNF-superoxide-MMP axis of events is in fact activated in heart disease in vivo. Wild-type and TNF(-/-) mice subjected to cardiac pressure overload revealed that TNF deficiency resulted in reduced superoxide levels, collagenase activities, PI3K activity, and fibrosis leading to attenuated cardiac dilation and dysfunction. Our study demonstrates that TNF triggers expression and activation of MMPs faster and stronger in cardiomyocytes than in cardiofibroblasts in a superoxide-dependent manner and via activation of PI3Kgamma, thereby contributing to adverse myocardial remodeling in disease. PMID- 20007454 TI - High concentrations of HGF inhibit skeletal muscle satellite cell proliferation in vitro by inducing expression of myostatin: a possible mechanism for reestablishing satellite cell quiescence in vivo. AB - Skeletal muscle regeneration and work-induced hypertrophy rely on molecular events responsible for activation and quiescence of resident myogenic stem cells, satellite cells. Recent studies demonstrated that hepatocyte growth factor (HGF) triggers activation and entry into the cell cycle in response to mechanical perturbation, and that subsequent expression of myostatin may signal a return to cell quiescence. However, mechanisms responsible for coordinating expression of myostatin after an appropriate time lag following activation and proliferation are not clear. Here we address the possible role of HGF in quiescence through its concentration-dependent negative-feedback mechanism following satellite cell activation and proliferation. When activated/proliferating satellite cell cultures were treated for 24 h beginning 48-h postplating with 10-500 ng/ml HGF, the percentage of bromodeoxyuridine-incorporating cells decreased down to a baseline level comparable to 24-h control cultures in a HGF dose-dependent manner. The high level HGF treatment did not impair the cell viability and differentiation levels, and cells could be reactivated by lowering HGF concentrations to 2.5 ng/ml, a concentration that has been shown to optimally stimulate activation of satellite cells in culture. Coaddition of antimyostatin neutralizing antibody could prevent deactivation and abolish upregulation of cyclin-dependent kinase (Cdk) inhibitor p21. Myostatin mRNA expression was upregulated with high concentrations of HGF, as demonstrated by RT-PCR, and enhanced myostatin protein expression and secretion were revealed by Western blots of the cell lysates and conditioned media. These results indicate that HGF could induce satellite cell quiescence by stimulating myostatin expression. The HGF concentration required (over 10-50 ng/ml), however, is much higher than that for activation, which is initiated by rapid release of HGF from its extracellular association. Considering that HGF is produced by satellite cells and spleen and liver cells in response to muscle damage, local concentrations of HGF bathing satellite cells may reach a threshold sufficient to induce myostatin expression. This time lag may delay action of the quiescence signaling program in proliferating satellite cells during initial phases of muscle regeneration followed by induction of quiescence in a subset of cells during later phases. PMID- 20007456 TI - From blood typing to a transport metabolon at a crossroad. Focus on "Ammonium dependent sodium uptake in mitochondrion-rich cells of medaka (Oryzias latipes) larvae". PMID- 20007455 TI - Tyrosine phosphorylation by Src within the cavity of the adenine nucleotide translocase 1 regulates ADP/ATP exchange in mitochondria. AB - Phosphorylation of adenine nucleotide translocator 1 (ANT1) at residue Y194, which is part of the aromatic ladder located within the lumen of the carrier, critically regulates mitochondrial metabolism. Recent data support the concept that members of the Src family of nonreceptor tyrosine kinases are constitutively present in mitochondria and key to regulation of mitochondrial function. Herein, we demonstrate that site mutations of ANT1 (Y190-->F190, Y194-->F194) mimicking dephosphorylation of the aromatic ladder resulted in loss of oxidative growth and ADP/ATP exchange activity in respiration-incompetent yeast expressing mutant chimeric yN-hANT1. ANT1 is phosphorylated at Y194 by the Src family kinase members Src and Lck, and increased phosphorylation is tightly linked to reduced cell injury in preconditioned protected vs. unprotected cardiac mitochondria. Molecular dynamics simulations find the overall structure of the phosphorylated ANT1 stable, but with an increased steric flexibility in the region of the aromatic ladder, matrix loop m2, and four helix-linking regions. Combined with an analysis of the putative cytosolic salt bridge network, we reason that the effect of phosphorylation on transport is likely due to an accelerated transition between the main two conformational states (c<-->m) of the carrier during the transport cycle. Since "aromatic signatures" are typical for other mitochondrial carrier proteins with important biological functions, our results may be more general and applicable to these carriers. PMID- 20007457 TI - Iron supply determines apical/basolateral membrane distribution of intestinal iron transporters DMT1 and ferroportin 1. AB - Intestinal iron absorption comprises the coordinated activity of the influx transporter divalent metal transporter 1 (DMT1) and the efflux transporter ferroportin (FPN). In this work, we studied the movement of DMT1 and FPN between cellular compartments as a function of iron supply. In rat duodenum, iron gavage resulted in the relocation of DMT1 to basal domains and the internalization of basolateral FPN. Considerable FPN was also found in apical domains. In Caco-2 cells, the apical-to-basal movement of cyan fluorescent protein-tagged DMT1 was complete 90 min after the addition of iron. Steady-state membrane localization studies in Caco-2 cells revealed that iron status determined the apical/basolateral membrane distribution of DMT1 and FPN. In agreement with the membrane distribution of the transporters, (55)Fe flux experiments revealed inward and outward iron fluxes at both membrane domains. Antisense oligonucleotides targeted to DMT1 or FPN inhibited basolateral iron uptake and apical iron efflux, respectively, indicating the participation of DMT1 and FPN in these fluxes. The fluxes were regulated by the iron supply; increased iron reduced apical uptake and basal efflux and increased basal uptake and apical efflux. These findings suggest a novel mechanism of regulation of intestinal iron absorption based on inward and outward fluxes at both membrane domains, and repositioning of DMT1 and FPN between membrane and intracellular compartments as a function of iron supply. This mechanism should be complementary to those based in the transcriptional or translational regulation of iron transport proteins. PMID- 20007458 TI - The role of the bed nucleus of the stria terminalis in learning to fear. PMID- 20007459 TI - Neuronal degeneration in a viral model of multiple sclerosis. PMID- 20007461 TI - AF4 is a critical regulator of the IGF-1 signaling pathway during Purkinje cell development. AB - Deregulation of the insulin-like growth factor 1 (IGF-1) signaling pathway is a recurrent finding in mouse models and human patients with cerebellar ataxia and thus represents a common pathological cascade in neuronal cell death that may be targeted for therapy. We have previously identified a point mutation in AF4, a transcription cofactor of RNA polymerase II elongation and chromatin remodeling, that causes progressive and highly specific Purkinje cell (PC) death in the ataxic mouse mutant robotic, leading to the accumulation of AF4 in PCs. Here we confirm that the spatiotemporal pattern of PC degeneration in the robotic cerebellum correlates with the specific profile of AF4 upregulation. To identify the underlying molecular pathways, we performed microarray gene expression analysis of PCs obtained by laser capture microdissection (LCM) at the onset of degeneration. Igf-1 was significantly downregulated in robotic PCs compared with wild-type controls before and throughout the degenerative process. Consistently, we observed a decrease in the activation of downstream signaling molecules including type 1 IGF receptor (IGF-1R) and the extracellular signal-regulated kinase (ERK) 1 and ERK2. Chromatin immunoprecipitation confirmed that Igf-1 is a direct and the first validated target of the AF4 transcriptional regulatory complex, and treatment of presymptomatic robotic mice with IGF-1 indeed markedly delayed the progression of PC death. This study demonstrates that small changes in the levels of a single transcriptional cofactor can deleteriously affect normal cerebellum function and opens new avenues of research for the manipulation of the IGF-1 pathway in the treatment of cerebellar ataxia in humans. PMID- 20007460 TI - The lysosomal sialic acid transporter sialin is required for normal CNS myelination. AB - Salla disease and infantile sialic acid storage disease are autosomal recessive lysosomal storage disorders caused by mutations in the gene encoding sialin, a membrane protein that transports free sialic acid out of the lysosome after it is cleaved from sialoglycoconjugates undergoing degradation. Accumulation of sialic acid in lysosomes defines these disorders, and the clinical phenotype is characterized by neurodevelopmental defects, including severe CNS hypomyelination. In this study, we used a sialin-deficient mouse to address how loss of sialin leads to the defect in myelination. Behavioral analysis of the sialin(-/-) mouse demonstrates poor coordination, seizures, and premature death. Analysis by histology, electron microscopy, and Western blotting reveals a decrease in myelination of the CNS but normal neuronal cytoarchitecture and normal myelination of the PNS. To investigate potential mechanisms underlying CNS hypomyelination, we studied myelination and oligodendrocyte development in optic nerves. We found reduced numbers of myelinated axons in optic nerves from sialin( /-) mice, but the myelin that was present appeared grossly normal. Migration and density of oligodendrocyte precursor cells were normal; however, a marked decrease in the number of postmitotic oligodendrocytes and an associated increase in the number of apoptotic cells during the later stages of myelinogenesis were observed. These findings suggest that a defect in maturation of cells in the oligodendrocyte lineage leads to increased apoptosis and underlies the myelination defect associated with sialin loss. PMID- 20007462 TI - Phospholipase C beta 4 in the medial septum controls cholinergic theta oscillations and anxiety behaviors. AB - Anxiety is among the most prevalent and costly diseases of the CNS, but its underlying mechanisms are not fully understood. Although attenuated theta rhythms have been observed in human subjects with increased anxiety, no study has been done on the possible physiological link between these two manifestations. We found that the mutant mouse for phospholipase C beta 4 (PLC-beta 4(-/-)) showed attenuated theta rhythm and increased anxiety, presenting the first animal model for the human condition. PLC-beta 4 is abundantly expressed in the medial septum, a region implicated in anxiety behavior. RNA interference-mediated PLC-beta 4 knockdown in the medial septum produced a phenotype similar to that of PLC-beta 4(-/-) mice. Furthermore, increasing cholinergic signaling by administering an acetylcholinesterase inhibitor cured the anomalies in both cholinergic theta rhythm and anxiety behavior observed in PLC-beta 4(-/-) mice. These findings suggest that (1) PLC-beta 4 in the medial septum is involved in controlling cholinergic theta oscillation and (2) cholinergic theta rhythm plays a critical role in suppressing anxiety. We propose that defining the cholinergic theta rhythm profile may provide guidance in subtyping anxiety disorders in humans for more effective diagnosis and treatments. PMID- 20007463 TI - Associatively learned representations of taste outcomes activate taste-encoding neural ensembles in gustatory cortex. AB - Through learning processes, cues associated with emotionally salient reinforcing outcomes can come to act as substitutes for the reinforcer itself. According to one account of this phenomenon, the predictive cue associatively elicits a representation of the expected outcome by reactivating cells responsible for encoding features of the primary reinforcer. We tested this hypothesis by examining the role of neural ensembles in gustatory cortex (GC) during receipt of gustatory stimuli (sucrose and water) and cues associated with those stimuli using the immediate early genes (IEGs) Arc and Homer1a. Because these plasticity related IEGs are expressed in the neuronal nucleus 5 and 30 min, respectively, after salient events, we examined how individual neurons encoded these stimuli in two separate behavioral epochs. In experiment 1, we showed that tasting identical sucrose solutions, but not tasteless water, in the two epochs increased both IEG activity and the degree of overlap between neural ensembles in GC. In experiment 2, odor cues associated with sucrose, but not water, evoked potentiation of IEG activity in GC similar to sucrose itself. Surprisingly, lesions of the basolateral amygdala had minimal effects on associative encoding in GC. Finally, these associatively driven representations of sucrose appeared to be outcome specific, as neural ensembles that were activated by the sucrose-associated cue were also activated by sucrose itself. This degree of overlap between associative and primary taste activity at the ensemble level suggests that GC neurons encode important information about anticipated outcomes. Such representations may provide outcome-specific information for guiding goal-directed behavior. PMID- 20007464 TI - Cytotoxic CD8+ T cell-neuron interactions: perforin-dependent electrical silencing precedes but is not causally linked to neuronal cell death. AB - Cytotoxic CD8(+) T cells are considered important effector cells contributing to neuronal damage in inflammatory and degenerative CNS disorders. Using time-lapse video microscopy and two-photon imaging in combination with whole-cell patch clamp recordings, we here show that major histocompatibility class I (MHC I) restricted neuronal antigen presentation and T cell receptor specificity determine CD8(+) T-cell locomotion and neuronal damage in culture and hippocampal brain slices. Two separate functional consequences result from a direct cell-cell contact between antigen-presenting neurons and antigen-specific CD8(+) T cells. (1) An immediate impairment of electrical signaling in single neurons and neuronal networks occurs as a result of massive shunting of the membrane capacitance after insertion of channel-forming perforin (and probably activation of other transmembrane conductances), which is paralleled by an increase of intracellular Ca(2+) levels (within <10 min). (2) Antigen-dependent neuronal apoptosis may occur independently of perforin and members of the granzyme B cluster (within approximately 1 h), suggesting that extracellular effects can substitute for intracellular delivery of granzymes by perforin. Thus, electrical silencing is an immediate consequence of MHC I-restricted interaction of CD8(+) T cells with neurons. This mechanism is clearly perforin-dependent and precedes, but is not causally linked, to neuronal cell death. PMID- 20007465 TI - Olfactory impairment predicts brain atrophy in Parkinson's disease. AB - Olfactory dysfunction is a frequent nonmotor symptom in idiopathic Parkinson's disease (PD) and may be considered as an early clinical feature of the disease preceding motor symptoms by years. According to recent neuropathological staging concepts, impaired olfaction is assumed to indicate an early pathological process and might be associated with structural changes in the brain. A morphometric analysis of magnetic resonance images [voxel-based morphometry (VBM)] was used to investigate gray matter atrophy related to psychophysically measured scores of olfactory function in early PD patients (n = 15, median Hoehn and Yahr stage 1.5), moderately advanced PD patients (n = 12, median Hoehn and Yahr stage 2.5), and age-matched healthy controls (n = 17). In PD patients, but not in controls, cortical atrophy in olfactory-related brain regions correlated specifically with olfactory dysfunction. Positive correlations between olfactory performance and gray matter volume were observed in the right piriform cortex in early PD patients and in the right amygdala in moderately advanced patients. The results provided first evidence that olfactory dysfunction in PD is related to atrophy in olfactory-eloquent regions of the limbic and paralimbic cortex. In addition, olfactory-correlated atrophy in these brain regions is consistent with the assumption that olfactory impairment as an early symptom of PD is likely to be associated with extranigral pathology. PMID- 20007466 TI - Cav1.3 channel voltage dependence, not Ca2+ selectivity, drives pacemaker activity and amplifies bursts in nigral dopamine neurons. AB - Ca(v)1.3 (alpha 1D) L-type Ca(2+) channels have been implicated in substantia nigra (SN) dopamine (DA) neuron pacemaking and vulnerability to Parkinson's disease. These effects may arise from the depolarizing current and cytoplasmic Ca(2+) elevation produced by Ca(v)1.3 channels at subthreshold membrane potentials. However, the assumption that the Ca(2+) selectivity of Ca(v)1.3 channels is essential has not been tested. In this study the properties of SN DA neuron L-type Ca(2+) channels responsible for driving pacemaker activity in juvenile rat brain slices were probed by replacing native channels blocked with the dihydropyridine nimodipine with virtual channels generated by dynamic clamp. Surprisingly, virtual L-type channels that mimic native and recombinant Ca(v)1.3 channels supported pacemaker activity even though dynamic clamp currents are not carried by Ca(2+). This effect is specific because pacemaker activity could not be restored by tonic current injection, virtual nonselective leak channels or virtual NMDA receptors, which share with L-type channels a negative slope conductance region in their current-voltage (I-V) curve. Altering virtual channels showed that the production of pacemaker activity depended on the characteristic voltage dependence of DA neuron L-type channels, while activation kinetics and reversal potential were not critical parameters. Virtual L-type channels also supported slow oscillatory potentials and enhanced firing rate during evoked bursts. Thus, Ca(v)1.3 channel voltage dependence, rather than Ca(2+) selectivity, drives pacemaker activity and amplifies bursts in SN DA neurons. PMID- 20007468 TI - cAMP-dependent axon guidance is distinctly regulated by Epac and protein kinase A. AB - cAMP is a key mediator of a number of molecules that induce growth cone chemotaxis, including netrin-1 and myelin-associated glycoprotein (MAG). Endogenous neuronal cAMP levels decline during development, and concomitantly axonal growth cones switch their response to cAMP-dependent guidance cues from attraction to repulsion. The mechanisms by which cAMP regulates these polarized growth cone responses are unknown. We report that embryonic growth cone attraction to gradients of cAMP, netrin-1, or MAG is mediated by Epac. Conversely, the repulsion conferred by MAG or netrin-1 on adult growth cones is mediated by protein kinase A (PKA). Furthermore, fluorescence resonance energy transfer reveals that netrin-1 distinctly activates Epac in embryonic growth cones but PKA in postnatal neurons. Our results suggest that cAMP mediates growth cone attraction or repulsion by distinctly activating Epac or PKA, respectively. Moreover, we propose that the developmental switch in growth cone response to gradients of cAMP-dependent guidance cues from attraction to repulsion is the result of a switch from Epac- to PKA-mediated signaling pathways. PMID- 20007467 TI - Millisecond timescale disinhibition mediates fast information transmission through an avian basal ganglia loop. AB - Avian song learning shares striking similarities with human speech acquisition and requires a basal ganglia (BG)-thalamo-cortical circuit. Information processing and transmission speed in the BG is thought to be limited by synaptic architecture of two serial inhibitory connections. Propagation speed may be critical in the avian BG circuit given the temporally precise control of musculature during vocalization. We used electrical stimulation of the cortical inputs to the BG to study, with fine time resolution, the functional connectivity within this network. We found that neurons in thalamic and cortical nuclei that are not directly connected with the stimulated area can respond to the stimulation with extremely short latencies. Through pharmacological manipulations, we trace this property back to the BG and show that the cortical stimulation triggers fast disinhibition of the thalamic neurons. Surprisingly, feedforward inhibition mediated by striatal inhibitory neurons onto BG output neurons sometimes precedes the monosynaptic excitatory drive from cortical afferents. The fast feedforward inhibition lengthens a single interspike interval in BG output neurons by just a few milliseconds. This short delay is sufficient to drive a strong, brief increase in firing probability in the target thalamic neurons, evoking short-latency responses. By blocking glutamate receptors in vivo, we show that thalamic responses do not appear to rely on excitatory drive, and we show in a theoretical model that they could be mediated by postinhibitory rebound properties. Such fast signaling through disinhibition and rebound may be a crucial specialization for learning of rapid and temporally precise motor acts such as vocal communication. PMID- 20007469 TI - Dramatic reduction of PrP C level and glycosylation in peripheral nerves following PrP knock-out from Schwann cells does not prevent transmissible spongiform encephalopathy neuroinvasion. AB - Expression of the prion protein (PrP(C)) is a requirement for host susceptibility to the transmissible spongiform encephalopathies (TSEs) and thought to be necessary for the replication and transport of the infectious agent. The mechanism of TSE neuroinvasion is not fully understood, although the routing of infection has been mapped through the peripheral nervous system (PNS) and Schwann cells have been implicated as a potential conduit for transport of the TSE infectious agent. To address whether Schwann cells are a requirement for spread of the TSE agent from the site of infection to the CNS, PrP(C) expression was selectively removed from Schwann cells in vivo. This dramatically reduced total PrP(C) within peripheral nerves by 90%, resulting in the selective loss of glycosylated PrP(C) species. Despite this, 139A and ME7 mouse-passaged scrapie agent strains were efficiently replicated and transported to the CNS following oral and intraperitoneal exposure. Thus, the myelinating glial cells within the PNS do not appear to play a significant role in TSE neuroinvasion. PMID- 20007470 TI - Four projection streams from primate V1 to the cytochrome oxidase stripes of V2. AB - In the primate visual system, areas V1 and V2 distribute information they receive from the retina to all higher cortical areas, sorting this information into dorsal and ventral streams. Therefore, knowledge of the organization of projections between V1 and V2 is crucial to understand how the cortex processes visual information. In primates, parallel output pathways from V1 project to distinct V2 stripes. The traditional tripartite division of V1-to-V2 projections was recently replaced by a bipartite scheme, in which thin stripes receive V1 inputs from blob columns, and thick and pale stripes receive common input from interblob columns. Here, we demonstrate that thick and pale stripes, instead, receive spatially segregated V1 inputs and that the interblob is partitioned into two compartments: the middle of the interblob projecting to pale stripes and the blob/interblob border region projecting to thick stripes. Double-labeling experiments further demonstrate that V1 cells project to either thick or pale stripes, but rarely to both. We also find laminar specialization of V1 outputs, with layer 4B contributing projections mainly to thick stripes, and no projections to one set of pale stripes. These laminar differences suggest different contribution of magno, parvo, and konio inputs to each V1 output pathway. These results provide a new foundation for parallel processing models of the visual system by demonstrating four V1-to-V2 pathways: blob columns-to-thin stripes, blob/interblob border columns-to-thick stripes, interblob columns-to pale(lateral) stripes, layer 2/3-4A interblobs-to-pale(medial) stripes. PMID- 20007471 TI - Brain-derived neurotrophic factor reduces amyloidogenic processing through control of SORLA gene expression. AB - Sorting protein-related receptor with A-type repeats (SORLA) is a major risk factor in cellular processes leading to Alzheimer's disease (AD). It acts as sorting receptor for the amyloid precursor protein (APP) that regulates intracellular trafficking and processing into amyloidogenic-beta peptides (A beta). Overexpression of SORLA in neurons reduces while inactivation of gene expression (as in knock-out mouse models) accelerates amyloidogenic processing and senile plaque formation. The current study aimed at identifying molecular pathways that control SORLA gene transcription in vivo and that may contribute to low levels of receptor expression in the brain of patients with AD. Using screening approaches in primary neurons, we identified brain-derived neurotrophic factor (BDNF) as a major inducer of Sorla that activates receptor gene transcription through the ERK (extracellular regulated kinase) pathway. In line with a physiological role as regulator of Sorla, expression of the receptor is significantly impaired in mouse models with genetic (Bdnf(-/-)) or disease related loss of BDNF activity in the brain (Huntington's disease). Intriguingly, exogenous application of BDNF reduced A beta production in primary neurons and in the brain of wild-type mice in vivo, but not in animals genetically deficient for Sorla. These findings demonstrate that the beneficial effects ascribed to BDNF in APP metabolism act through induction of Sorla that encodes a negative regulator of neuronal APP processing. PMID- 20007472 TI - Functional excitatory microcircuits in neonatal cortex connect thalamus and layer 4. AB - The functional connectivity of the cerebral cortex is shaped by experience during development, especially during a critical period early in life. In the prenatal and neonatal cortex, transient neuronal circuits are formed by a population of subplate neurons (SPNs). However, SPNs are absent in the adult cortex. While SPNs are crucial for normal development of the cerebral cortex and of thalamocortical synapses, little is known about how they are integrated in the developing thalamocortical circuit. We therefore investigated SPNs in vitro in thalamocortical slices of A1 and medial geniculate nucleus (MGN) in mouse from postnatal day 1 (P1) to P13. We found that SPNs can fire action potentials at P1 and that their intrinsic membrane properties are mature after P5. We find that SPNs receive functional excitatory inputs from the MGN as early as P2. The MGN projections to SPNs strengthen between P2 and P13 and are capable of inducing action potentials in SPNs. Selective activation of SPNs by photostimulation produced EPSCs in layer 4 neurons, demonstrating a functional excitatory connection. Thus, SPNs are tightly integrated into the developing thalamocortical circuit and would be a reliable relay of early spontaneous and sound-evoked activity. The role of SPNs in development likely results from their strong excitatory projection to layer 4, which might function to regulate activity dependent processes that enable mechanisms required for the functional maturation and plasticity of the developing cortex and thereby contribute to the development of normal cortical organization. PMID- 20007473 TI - No Nogo66- and NgR-mediated inhibition of regenerating axons in the zebrafish optic nerve. AB - In contrast to mammals, lesioned axons in the zebrafish (ZF) optic nerve regenerate and restore vision. This correlates with the absence of the NogoA specific N-terminal domains from the ZF nogo/rtn-4 (reticulon-4) gene that inhibits regeneration in mammals. However, mammalian nogo/rtn-4 carries a second inhibitory C-terminal domain, Nogo-66, being 70% identical with ZF-Nogo66. The present study examines, (1) whether ZF-Nogo66 is inhibitory and effecting similar signaling pathways upon Nogo66-binding to the Nogo66 receptor NgR and its coreceptors, and (2) whether Rat-Nogo66 on fish, and ZF-Nogo66 on mouse neurons, cause inhibition via NgR. Our results from "outgrowth, collapse and contact assays" suggest, surprisingly, that ZF-Nogo66 is growth-permissive for ZF and mouse neurons, quite in contrast to its Rat-Nogo66 homolog which inhibits growth. The opposite effects of ZF- and Rat-Nogo66 are, in both fish and mouse, transmitted by GPI (glycosylphosphatidylinositol)-anchored receptors, including NgR. The high degree of sequence homology in the predicted binding site is consistent with the ability of ZF- and mammalian-Nogo66 to bind to NgRs of both species. Yet, Rat-Nogo66 elicits phosphorylation of the downstream effector cofilin whereas ZF-Nogo66 has no influence on cofilin phosphorylation--probably because of significantly different Rat- versus ZF-Nogo66 sequences outside of the receptor-binding region effecting, by speculation, recruitment of a different set of coreceptors or microdomain association of NgR. Thus, not only was the NogoA specific domain lost in fish, but Nogo66, the second inhibitory domain in mammals, and its signaling upon binding to NgR, was modified so that ZF-Nogo/RTN 4 does not impair axon regeneration. PMID- 20007474 TI - Atypical functional properties of GluK3-containing kainate receptors. AB - The properties of synaptic receptors determine their mode of action at presynaptic and postsynaptic loci. Here, we investigated the atypical biophysical properties of GluK3-containing kainate receptors, which contribute to presynaptic facilitation at hippocampal mossy fiber synapses. We show, using fast glutamate applications on outside-out patches and kinetic modeling, that the low sensitivity of GluK3 receptors for glutamate is attributable to fast desensitization of partially bound receptors. Consequently, these receptors can only be activated by fast transients of high glutamate concentration. In addition, GluK3 receptors are very sensitive to voltage-dependent block by intracellular spermine that precludes activation of substantial currents at potentials positive to -50 mV. Two specific residues within the channel pore define this high-affinity site. Finally, GluK3 are calcium permeable in the same way as unedited GluK2 receptors. These receptors present unique properties among AMPA/kainate receptors that could reflect a specialized presynaptic function. PMID- 20007475 TI - Brain-derived neurotrophic factor signaling in the HVC is required for testosterone-induced song of female canaries. AB - Testosterone-induced singing in songbirds is thought to involve testosterone dependent morphological changes that include angiogenesis and neuronal recruitment into the HVC, a central part of the song control circuit. Previous work showed that testosterone induces the production of vascular endothelial growth factor (VEGF) and its receptor (VEGFR2 tyrosine kinase), which in turn leads to an upregulation of brain-derived neurotrophic factor (BDNF) production in HVC endothelial cells. Here we report for the first time that systemic inhibition of the VEGFR2 tyrosine kinase is sufficient to block testosterone induced song in adult female canaries, despite sustained androgen exposure and the persistence of the effects of testosterone on HVC morphology. Expression of exogenous BDNF in HVC, induced locally by in situ transfection, reversed the VEGFR2 inhibition-mediated blockade of song development, thereby restoring the behavioral phenotype associated with androgen-induced song. The VEGFR2-inhibited, BDNF-treated females developed elaborate male-like song that included large syllable repertoires and high syllable repetition rates, features known to attract females. Importantly, although functionally competent new neurons were recruited to HVC after testosterone treatment, the time course of neuronal addition appeared to follow BDNF-induced song development. These findings indicate that testosterone-associated VEGFR2 activity is required for androgen induced song in adult songbirds and that the behavioral effects of VEGFR2 inhibition can be rescued by BDNF within the adult HVC. PMID- 20007476 TI - Distinct dose-dependent cortical neuronal migration and neurite extension defects in Lis1 and Ndel1 mutant mice. AB - Haploinsufficiency of LIS1 results in lissencephaly, a human neuronal migration disorder. LIS1 is a microtubule- (MT) and centrosome- [microtubule organizing center (MTOC)] associated protein that regulates nucleokinesis via the regulation of dynein motor function and localization. NDEL1 (NudE isoform, NudE like) interacts with LIS1/dynein complex, and is phosphorylated by CDK5/P35. Previous reports using siRNA-mediated knock-down demonstrated similar critical roles for LIS1 and NDEL1 during neuronal migration, but neuronal migration has not been studied in genetic mutants for Lis1 and Ndel1 where protein levels are uniform in all cells. Brains from mice with complete loss of Lis1 and Ndel1 displayed severe cortical layering and hippocampal defects, but Lis1 mutants had more severe defects. Neuronal migration speed was reduced and neurite lengths were elongated in proportion to the reduction of LIS1 and NDEL1 protein levels in embryonic day 14.5 mutant cortical slices compared to wild type, using two-photon confocal time lapse videomicroscopy. Additionally, mice with 35% of wild-type NDEL1 levels displayed diverse branched migration modes with multiple leading processes, suggesting defects in adhesion and/or polarity. Complete loss of Lis1 or Ndel1 resulted in the total inhibition of nuclear movement in cortical slice assays, and in neurosphere assays, the percentage of migrating neurons with correctly polarized MTOC location was significantly reduced while nuclear-centrosomal distance was extended. Neurite lengths were increased after complete loss Ndel1 but reduced after complete loss of Lis1. Thus, Lis1 and Ndel1 are essential for normal cortical neuronal migration, neurite outgrowth, and function of the MTOC in a dose-dependent manner. PMID- 20007477 TI - Cellular mechanisms underlying burst firing in substantia nigra dopamine neurons. AB - Burst firing of substantia nigra dopamine (SN DA) neurons is believed to represent an important teaching signal that instructs synaptic plasticity and associative learning. However, the mechanisms through which synaptic excitation overcomes the limiting effects of somatic Ca(2+)-dependent K(+) current to generate burst firing are controversial. Modeling studies suggest that synaptic excitation sufficiently amplifies oscillatory dendritic Ca(2+) and Na(+) channel currents to lead to the initiation of high-frequency firing in SN DA neuron dendrites. To test this model, visually guided compartment-specific patch-clamp recording and ion channel manipulation were applied to rodent SN DA neurons in vitro. As suggested previously, the axon of SN DA neurons was typically found to originate from a large-diameter dendrite that was proximal to the soma. However, in contrast to the predictions of the model, (1) somatic current injection generated firing that was similar in frequency and form to burst firing in vivo, (2) the efficacy of glutamatergic excitation was inversely related to the distance of excitation from the axon, (3) pharmacological blockade or genetic deletion of Ca(2+) channels did not prevent high-frequency firing, (4) action potential bursts were invariably detected first at sites that were proximal to the axon, and (5) pharmacological blockade of Na(+) channels in the vicinity of the axon/soma but not dendritic excitation impaired burst firing. Together, these data suggest that SN DA neurons integrate their synaptic input in a more conventional manner than was hypothesized previously. PMID- 20007478 TI - NFIL3 and cAMP response element-binding protein form a transcriptional feedforward loop that controls neuronal regeneration-associated gene expression. AB - Successful regeneration of damaged neurons depends on the coordinated expression of neuron-intrinsic genes. At present however, there is no comprehensive view of the transcriptional regulatory mechanisms underlying neuronal regeneration. We used high-content cellular screening to investigate the functional contribution of 62 transcription factors to regenerative neuron outgrowth. Ten transcription factors are identified that either increase or decrease neurite outgrowth. One of these, NFIL3, is specifically upregulated during successful regeneration in vivo. Paradoxically however, knockdown of NFIL3 and overexpression of dominant-negative NFIL3 both increase neurite outgrowth. Our data show that NFIL3, together with CREB, forms an incoherent feedforward transcriptional regulatory loop in which NFIL3 acts as a negative regulator of CREB-induced regeneration-associated genes. PMID- 20007479 TI - Activation of glycogen synthase kinase-3 beta is required for hyperdopamine and D2 receptor-mediated inhibition of synaptic NMDA receptor function in the rat prefrontal cortex. AB - The interactions between dopamine and glutamate systems play an essential role in normal brain functions and neuropsychiatric disorders. The mechanism of NMDA receptor regulation through high concentrations of dopamine, however, remains unclear. Here, we show the signaling pathways involved in hyperdopaminergic regulation of NMDA receptor functions in the prefrontal cortex by incubating cortical slices with high concentration of dopamine or administering dopamine reuptake inhibitor 1-(2-[bis-(4-fluorophenyl)methoxy]ethyl)- 4-(3 phenylpropyl)piperazine (GBR12909) in vivo. We found that, under both conditions, the synaptic NMDA receptor-mediated currents were significantly attenuated by excessive dopamine stimulation through activation of D(2) receptors. Furthermore, high dose of dopamine failed to affect NMDA receptor-mediated currents after blockade of NR2B subunits but triggered a dynamin-dependent endocytosis of NMDA receptors. The high-dose dopamine/D(2) receptor-mediated suppression of NMDA receptors was involved in the increase of glycogen synthase kinase-3beta (GSK 3beta) activity, which in turn phosphorylates beta-catenin and disrupts beta catenin-NR2B interaction, but was dependent on neither Gq11 nor PLC (phospholipase C). Moreover, the hyperdopamine induced by GBR12909 significantly decreased the expression of both surface and intracellular NR2B proteins, as well as NR2B mRNA levels, suggesting an inhibition of protein synthesis. These effects were, however, completely reversed by administration of either GSK-3beta inhibitor or D(2) receptor antagonist. These results therefore suggest that GSK 3beta is required for the hyperdopamine/D(2) receptor-mediated inhibition of NMDA receptors in the prefrontal neurons and these actions may underlie D(2) receptor mediated psychostimulant effects and hyperdopamine-dependent behaviors in the brain. PMID- 20007480 TI - Temporal envelope of time-compressed speech represented in the human auditory cortex. AB - Speech comprehension relies on temporal cues contained in the speech envelope, and the auditory cortex has been implicated as playing a critical role in encoding this temporal information. We investigated auditory cortical responses to speech stimuli in subjects undergoing invasive electrophysiological monitoring for pharmacologically refractory epilepsy. Recordings were made from multicontact electrodes implanted in Heschl's gyrus (HG). Speech sentences, time compressed from 0.75 to 0.20 of natural speaking rate, elicited average evoked potentials (AEPs) and increases in event-related band power (ERBP) of cortical high frequency (70-250 Hz) activity. Cortex of posteromedial HG, the presumed core of human auditory cortex, represented the envelope of speech stimuli in the AEP and ERBP. Envelope following in ERBP, but not in AEP, was evident in both language dominant and -nondominant hemispheres for relatively high degrees of compression where speech was not comprehensible. Compared to posteromedial HG, responses from anterolateral HG-an auditory belt field-exhibited longer latencies, lower amplitudes, and little or no time locking to the speech envelope. The ability of the core auditory cortex to follow the temporal speech envelope over a wide range of speaking rates leads us to conclude that such capacity in itself is not a limiting factor for speech comprehension. PMID- 20007481 TI - Altered sleep homeostasis after restraint stress in 5-HTT knock-out male mice: a role for hypocretins. AB - Restraint stress produces changes in the sleep pattern that are mainly characterized by a delayed increase in rapid eye movement sleep (REMS) amounts. Because the serotonin (5-HT) and the hypocretin (hcrt) systems that regulate REMS are interconnected, we used mutant mice deficient in the 5-HT transporter (5-HTT( /-)) to examine the role of 5-HT and hcrt neurotransmissions in the sleep response to stress. In contrast to wild-type mice, restraint stress did not induce a delayed increase in REMS amounts in 5-HTT(-/-) mice, indicating impaired sleep homeostasis in mutants. However, pharmacological blockade of the hcrt type 1 receptor (hcrt-R1) before restraint stress restored the REMS increase in 5-HTT( /-) mice. In line with this finding, 5-HTT(-/-) mutants displayed after restraint stress higher long-lasting activation of hypothalamic preprohcrt neurons than wild-type mice and elevated levels of the hcrt-1 peptide and the hcrt-R1 mRNA in the anterior raphe area. Thus, hypocretinergic neurotransmission was enhanced by stress in 5-HTT(-/-) mice. Furthermore, in 5-HTT(-/-) but not wild-type mice, hypothalamic levels of the 5-HT metabolite 5-hydroxyindole acetic acid significantly increased after restraint stress, indicating a marked enhancement of serotonergic neurotransmission in mutants. Altogether, our data show that increased serotonergic -and in turn hypocretinergic- neurotransmissions exert an inhibitory influence on stress-induced delayed REMS. We propose that the direct interactions between hcrt neurons in the hypothalamus and 5-HT neurons in the anterior raphe nuclei account, at least in part, for the adaptive sleep wakefulness regulations triggered by acute stress. PMID- 20007482 TI - Neuromodulation at single presynaptic boutons of cerebellar parallel fibers is determined by bouton size and basal action potential-evoked Ca transient amplitude. AB - Most presynaptic terminals in the brain contain G-protein-coupled receptors that function to reduce action potential-evoked neurotransmitter release. These neuromodulatory receptors, including those for glutamate, GABA, endocannabinoids, and adenosine, exert a substantial portion of their effect by reducing evoked presynaptic Ca(2+) transients. Many axons form synapses with multiple postsynaptic neurons, but it is unclear whether presynaptic attenuation in these synapses is homogeneous, as suggested by population-level Ca(2+) imaging. We loaded Ca(2+)-sensitive dyes into cerebellar parallel fiber axons and imaged action potential-evoked Ca(2+) transients in individual presynaptic boutons with application of three different neuromodulators and found that adjacent boutons on the same axon showed striking heterogeneity in their strength of attenuation. Moreover, attenuation was predicted by bouton size or basal Ca(2+) response: smaller boutons were more sensitive to adenosine A1 agonist but less sensitive to CB1 agonist, while boutons with high basal action potential-evoked Ca(2+) transient amplitude were more sensitive to mGluR4 agonist. These results suggest that boutons within brief segment of a single parallel fiber axon can have different sensitivities toward neuromodulators and may have different capacities for both short-term and long-term plasticities. PMID- 20007483 TI - Neuronal avalanches imply maximum dynamic range in cortical networks at criticality. AB - Spontaneous neuronal activity is a ubiquitous feature of cortex. Its spatiotemporal organization reflects past input and modulates future network output. Here we study whether a particular type of spontaneous activity is generated by a network that is optimized for input processing. Neuronal avalanches are a type of spontaneous activity observed in superficial cortical layers in vitro and in vivo with statistical properties expected from a network operating at "criticality." Theory predicts that criticality and, therefore, neuronal avalanches are optimal for input processing, but until now, this has not been tested in experiments. Here, we use cortex slice cultures grown on planar microelectrode arrays to demonstrate that cortical networks that generate neuronal avalanches benefit from a maximized dynamic range, i.e., the ability to respond to the greatest range of stimuli. By changing the ratio of excitation and inhibition in the cultures, we derive a network tuning curve for stimulus processing as a function of distance from criticality in agreement with predictions from our simulations. Our findings suggest that in the cortex, (1) balanced excitation and inhibition establishes criticality, which maximizes the range of inputs that can be processed, and (2) spontaneous activity and input processing are unified in the context of critical phenomena. PMID- 20007484 TI - Dynamic reweighting of visual and vestibular cues during self-motion perception. AB - The perception of self-motion direction, or heading, relies on integration of multiple sensory cues, especially from the visual and vestibular systems. However, the reliability of sensory information can vary rapidly and unpredictably, and it remains unclear how the brain integrates multiple sensory signals given this dynamic uncertainty. Human psychophysical studies have shown that observers combine cues by weighting them in proportion to their reliability, consistent with statistically optimal integration schemes derived from Bayesian probability theory. Remarkably, because cue reliability is varied randomly across trials, the perceptual weight assigned to each cue must change from trial to trial. Dynamic cue reweighting has not been examined for combinations of visual and vestibular cues, nor has the Bayesian cue integration approach been applied to laboratory animals, an important step toward understanding the neural basis of cue integration. To address these issues, we tested human and monkey subjects in a heading discrimination task involving visual (optic flow) and vestibular (translational motion) cues. The cues were placed in conflict on a subset of trials, and their relative reliability was varied to assess the weights that subjects gave to each cue in their heading judgments. We found that monkeys can rapidly reweight visual and vestibular cues according to their reliability, the first such demonstration in a nonhuman species. However, some monkeys and humans tended to over-weight vestibular cues, inconsistent with simple predictions of a Bayesian model. Nonetheless, our findings establish a robust model system for studying the neural mechanisms of dynamic cue reweighting in multisensory perception. PMID- 20007486 TI - Erlotinib resistance in mouse models of epidermal growth factor receptor-induced lung adenocarcinoma. AB - Seventy-five percent of lung adenocarcinomas with epidermal growth factor receptor (EGFR) mutations respond to treatment with the tyrosine kinase inhibitors (TKIs) gefitinib and erlotinib; however, drug-resistant tumors eventually emerge. In 60% of cases, resistant tumors carry a secondary mutation in EGFR (T790M), amplification of MET, or both. Here, we describe the establishment of erlotinib resistance in lung tumors, which were induced by mutant EGFR, in transgenic mice after multiple cycles of drug treatment; we detect the T790M mutation in five out of 24 tumors or Met amplification in one out of 11 tumors in these mice. This preclinical mouse model, therefore, recapitulates the molecular changes responsible for resistance to TKIs in human tumors and holds promise for the discovery of additional mechanisms of drug resistance in lung cancer. PMID- 20007485 TI - Lack of host SPARC enhances vascular function and tumor spread in an orthotopic murine model of pancreatic carcinoma. AB - Utilizing subcutaneous tumor models, we previously validated SPARC (secreted protein acidic and rich in cysteine) as a key component of the stromal response, where it regulated tumor size, angiogenesis and extracellular matrix deposition. In the present study, we demonstrate that pancreatic tumors grown orthotopically in Sparc-null (Sparc(-/-)) mice are more metastatic than tumors grown in wild type (Sparc(+/+)) littermates. Tumors grown in Sparc(-/-) mice display reduced deposition of fibrillar collagens I and III, basement membrane collagen IV and the collagen-associated proteoglycan decorin. In addition, microvessel density and pericyte recruitment are reduced in tumors grown in the absence of host SPARC. However, tumors from Sparc(-/-) mice display increased permeability and perfusion, and a subsequent decrease in hypoxia. Finally, we found that tumors grown in the absence of host SPARC exhibit an increase in alternatively activated macrophages. These results suggest that increased tumor burden in the absence of host SPARC is a consequence of reduced collagen deposition, a disrupted vascular basement membrane, enhanced vascular function and an immune-tolerant, pro metastatic microenvironment. PMID- 20007488 TI - Developing a tool for systematic inclusion of non-academic factors in dental school admissions: towards building diversity in the dental workforce. AB - This retrospective study aimed to determine the ability of a self-designed evaluation instrument to identify applicants who should be invited for an interview and to assess applications including non-academic factors in a consistent and systematic manner. The instrument was constructed using information culled from the American Dental Education Association Associated American Dental Schools Application Service (ADEA AADSAS) applications including academic (DAT, GPA) and non-academic qualifications (work experience, volunteer and extracurricular activities, rural and ethnic background, essays). Weights were assigned to each item assessed. Using applications received during the 2006 and 2007 admissions cycles, the instrument identified who should have been invited for interview. Descriptive statistics and specificity and sensitivity tests were conducted. The instrument allowed for a systematic quantitative assessment of non-academic factors that was part of the overall evaluation of applicants. Results show differences in non-academic scores of minority compared to non-minority applicants, males compared to females, and those who were offered interviews compared to those not interviewed. It is possible that some applicants who were not offered interviews could have gained a chance to be looked at more closely on a second round of review if non-academic factors were considered. PMID- 20007489 TI - The impact of new dental schools on the dental workforce through 2022. AB - Following a wave of dental school closures from 1986 to 2001 and a perceived shortage of dentists, three new dental schools were established between 1997 and 2003, and eight more are in various stages of planning and development to open over the next decade. Conditions are moving rapidly, and several institutions have stated intentions to open new dental schools since this analysis. This article presents a supply-side analysis of the impact of the new schools on the effective dentist to population ratio, taking into account changes in graduation rates, retirement rate, population growth, productivity, and gender ratio of the profession. Demand-side factors including utilization, per patient expenditures, and case mix are addressed, as well as the implications of these changes on access to care and the future of the profession. Given approximately ten new schools, by 2022, an additional 8,233 graduates will have joined the workforce, or approximately three dentists per 100,000 people. Effective dentist to population ratios vary greatly depending on all of the factors addressed. Changes in productivity influence the effective ratio most significantly. Most probable scenarios for the dental workforce suggest a stable dentist to population ratio at minimum, with an increase likely given recent productivity changes. The increase in dentists will not noticeably improve access to care for low-income and rural populations absent additional public funding to support demand for these populations and concurrent measures to effect even distribution of dentists throughout the country. PMID- 20007487 TI - Rap1 and its effector KRIT1/CCM1 regulate beta-catenin signaling. AB - KRIT1, also called CCM1, is a member of a multiprotein complex that contains the products of the CCM2 and PDCD10 (also known as CCM3) loci. Heterozygous loss of any of the genes that encode these proteins leads to cerebral cavernous malformations (CCM), which are vascular lesions that are found in around 0.5% of humans. KRIT1 mediates the stabilization of beta-catenin-containing endothelial cell-cell junctions downstream of the Rap1 GTPase. Here, we report that Rap1 and KRIT1 are negative regulators of canonical beta-catenin signaling in mice and that hemizygous Krit1 deficiency exacerbates beta-catenin-driven pathologies. Depletion of endothelial KRIT1 caused beta-catenin to dissociate from vascular endothelial (VE)-cadherin and to accumulate in the nucleus with consequent increases in beta-catenin-dependent transcription. Activation of Rap1 inhibited beta-catenin-dependent transcription in confluent endothelial cells; this effect required the presence of intact cell-cell junctions and KRIT1. These effects of KRIT1 were not limited to endothelial cells; the KRIT1 protein was expressed widely and its depletion increased beta-catenin signaling in epithelial cells. Moreover, a reduction in KRIT1 expression also increased beta-catenin signaling in vivo. Hemizygous deficiency of Krit1 resulted in a ~1.5-fold increase in intestinal polyps in the Apc(Min/+) mouse, which was associated with increased beta-catenin-driven transcription. Thus, KRIT1 regulates beta-catenin signaling, and Krit1(+/-) mice are more susceptible to beta-catenin-driven intestinal adenomas. PMID- 20007490 TI - Dental students' attitudes towards digital textbooks. AB - Dental students at the Louisiana State University School of Dentistry (LSUSD) have participated in the VitalSource Bookshelf digital textbook program for the last four years. The Bookshelf, a collection of downloadable textbooks, includes the Contemporary Dental Library (CDL), a basic package of sixty books, plus additional titles selected by course directors. A survey was sent to students in the four LSUSD classes to assess satisfaction with the program. Eighty-two (34.2 percent) students completed the survey. In addition, exit interviews of graduating seniors contained a question on the Bookshelf. In both the survey and the exit interview, LSUSD students overwhelmingly expressed dissatisfaction with reading extensive amounts of text online and indicated a preference for printed books. Some students also resented paying for books in the CDL that were not required by instructors and felt that, considering how infrequently they used the Bookshelf, the cost was too high. PMID- 20007491 TI - Students' perceptions about pediatric dental behavior guidance in an undergraduate four-year dental curriculum. AB - Students' acceptability scores of pediatric dental behavior guidance techniques and clinical situations were measured with visual analog scales in a four-year dental curriculum. At the end of the curriculum, the highest scores were for positive reinforcement (94.7+/-4.7), use of nitrous oxide (93.1+/-7.5), stimulating the child's imagination (90.4+/-11.1), tell-show-do (90.4+/-10.0), distraction (89.7+/-11.6), use of euphemisms (88.3+/-14.4), voice control (86.2+/ 12.4), and promising a toy (83.6+/-17.7). The lowest scores were for showing a needle (23.1+/-20.9), treatment without local anesthetic (25.4+/-24.2), parent talking with the child during treatment (35.3+/-22.4), hand over mouth (37.1+/ 25.4), dentist/assistant being quiet (38.5+/-25.3), and not allowing the child to speak (38.6+/-26.8). Comparison of scores after only didactic education versus after clinical plus didactic education indicated a significant increase in acceptability for general anesthesia and significant decreases for situations involving the parent in the clinic. Ranking of scores before and after the dental curriculum showed the largest change in ranking for general anesthesia (+10), telling that the treatment may involve pain (-9), parent's presence (-8) or talking with the child during treatment (-10), and use of nitrous oxide (+7). Dental education has the potential to shape students' perceptions about pediatric dental behavior guidance. PMID- 20007492 TI - Do dental hygiene students fit the learning profile of the millennial student? AB - Differences in learning and the cultural context of our students' life experiences are important variables that faculty members need to understand in order to be effective in the classroom. Faculty members are finding that millennial students' approaches to learning are often vastly different from their own and as a result feel frustrated in their ability to help these students with their learning needs. Cultivating awareness of how today's dental hygiene student learns as well as the millennial learner profile can help faculty members address this educational challenge. The purpose of this study was to identify the learning styles of three groups of dental hygiene students and determine if they fit the learning profile of the millennial student as measured by the Learning Type Measure. Given this new generation of learners, it was hypothesized that dental hygiene students' learning style preferences would fit the learning profile of the millennial student. The Learning Type Measure was administered to 101 dental hygiene students at the University of Minnesota, University of Arizona, and Virginia Commonwealth University. The results from the study revealed that dental hygiene students do exhibit learning style preferences consistent with the millennial learner profile. PMID- 20007493 TI - Educational experiences and preparedness in dental anesthesia: five-year outcome assessment and conclusions. AB - A mail survey of 2003-07 dental school graduates was undertaken by the Department of Anesthesiology at the University of Pittsburgh School of Dental Medicine to assess the strengths and weaknesses of the predoctoral curriculum in anesthesia and to determine the preparedness of practicing dentists to provide anesthesia services for their dental patients. Subsets of the survey responses were created to specifically evaluate the effectiveness of an advanced selective program in sedation offered to approximately twenty third- and fourth-year predoctoral students. This fourteen-month Anesthesia Selective Program provides advanced didactic instruction and clinical experiences needed to establish competence in minimal to moderate sedation. Overall, graduates reported being best prepared in assessment of medical histories, physiology, and pharmacology, while being least prepared in oral sedation, intravenous sedation, and general anesthesia. For graduates currently in general practice, those who had participated in the Anesthesia Selective Program reported being better prepared in most subjects relating to anesthesia and patient care. Participants in the selective were also more likely to treat special needs patients in their private practices. Respondents' written comments indicated a desire for a greater number of clinical experiences involving sedation procedures within the predoctoral curriculum. This outcome assessment indicated that a greater emphasis should be placed on instruction and training experiences for enteral sedation within the predoctoral dental curriculum. Advanced training and increased clinical experiences in anesthesia may also be an effective means to better prepare graduates to assess medical histories, to manage medical emergencies, and to be willing to treat medically complex patients as well as patients with special health care needs. PMID- 20007494 TI - Dental practice network of U.S. dental schools. AB - As dental schools incorporate training in evidence-based dentistry (EBD) into their curricula, students must learn how to critically evaluate systematic reviews and meta-analyses. It is important that dental education in the United States support the American Dental Association's position statement on EBD, which defines "best evidence" as data obtained from all study designs. Given that much evidence is missing when EBD is derived from Cochrane Systematic Reviews' randomized clinical trials, we propose the creation of a dental practice network of U.S. dental schools. We developed an electronic clinical dentistry research database for EBD using Epi-Info (available at www.cdc.gov/epiinfo/downloads.htm). As a free, public use software, Epi-Info provides the foundation for the development of clinical research databases that can increase the research capacity through multisite studies designed to generate outcomes data on the effectiveness of dental treatment. The creation of a dental practice network of dental schools with their large number of patients would expand the research capacity for EBD practice and advance the EBD science regarding the effectiveness of dental treatment. The next step is to link clinical dental researchers/educators at multiple dental schools through a collaborative clinical research network, so that the findings can be applied to the EBD component of problem-based learning curricula of dental education. PMID- 20007495 TI - Dental school patients' understanding of informed consent. AB - The purpose of this qualitative exploratory study is to describe and analyze adult patients' understanding of informed consent, using the University of Pennsylvania School of Dental Medicine's consent form. Randomly sampled, anonymous patients at the school's clinic were asked to read the form and highlight any statements they found to be confusing, while also rating the form as "easy," "needing minor changes," or "needing major changes." This survey tested the hypothesis that the reading level of the form exceeds that of the average patient and thus, as education level increased, so would comprehensibility while the number of confusing statements would decrease. The results were analyzed for trends in gender and education level, and the statements proving most problematic to the largest number of patients were identified and analyzed. Contrary to the hypothesis, the most highly educated patients highlighted more statements, inspiring a new hypothesis that this occurred because of highly educated patients' ability to distinguish between the literal meaning of the words and their implications. Revisions were made to the identified aspects of the form, and new surveys were completed, with the result that the modifications were successful and the new hypothesis was supported. PMID- 20007496 TI - Factors involved in the choice of dentistry as an occupation by pernambuco dental students in Brazil. AB - The aim of this study was to determine the factors that lead to the choice of dentistry as an occupation among dental students in Pernambuco, Brazil. A total of 1,024 students were surveyed. All participants were enrolled in dentistry courses in the state of Pernambuco in 2008. Pearson's chi-square test and Fisher's exact test were used for the statistical analysis of the data, with the level of significance set at 5.0 percent. The results demonstrated that the choice of occupation was made primarily for economic reasons (73.5 percent), followed by vocational reasons (68.3 percent), professional reasons (67.8 percent), and personal reasons (56.6 percent). PMID- 20007497 TI - Medical problems among dental patients at the school of dentistry, the university of the West Indies. AB - This study ascertained demographic information and prevalence rates of medical problems among 571 new and consecutive dental patients attending the emergency clinic of the School of Dentistry, The University of the West Indies. Patients were interviewed by specially trained and calibrated dental students and interns. Various medical problems of patients were recorded in individual open-ended case sheets, maintaining their privacy and confidentiality. This information was then transcribed into a specially designed and pretested form. The data were entered into a Microsoft Excel spreadsheet and analyzed using the SPSS statistical package to obtain the prevalence rates of medical conditions, which were then cross-tabulated with gender, age, ethnicity, and other variables. Significance of differences, if any, was evaluated by chi-square test. In all, 303 medical conditions were encountered in 239 individuals, giving a prevalence rate of 42 percent. Hypertension (12.6 percent), diabetes (6.1 percent), asthma (5.8 percent), arthritis (4.7 percent), and various allergies (8.3 percent) constituted an important segment of the problems. Gender, ethnic, and age differences were also evident for some diseases. In this study-which was the first of this kind in the West Indies-vital information on medical problems among dental patients was obtained by interviews conducted by dental students and interns, forming an important part of their dental education. PMID- 20007500 TI - Developmental alteration of endocannabinoid retrograde signaling in the hippocampus. AB - Endocannabinoids are lipid derivatives that mediate paracrine and juxtacrine signaling between cells. In the hippocampal CA1 region, a retrograde endocannabinoid signal suppresses GABA release by acting on presynaptic cannabinoid receptor-1 (CB1) and can be functionally manifested as depolarization induced suppression of inhibition (DSI). In the present study, whole cell patch clamp recordings in hippocampal slices were made to examine DSI in rats from P7 P21. Robust DSI develops in rat hippocampus at postnatal ages greater than two weeks, but only modest DSI is observed in P7-9 rat. DSI in neonatal rats can be enhanced by activation of group I metabotropic glutamate receptors (mGluRs) or muscarinic acetylcholine receptors in those neonatal rats. The DSI is also enhanced by sustained low-frequency (1 Hz) stimulation (5 min). This stimulus enhanced DSI was prevented in the presence of 6-methyl-2-(phenylethynyl)-pyridine (10 microM), a group I mGluR antagonist. WIN55212-2, a synthetic CB1 agonist, produced a similar level of inhibition of GABAergic synaptic transmission at different postnatal time points. Therefore postsynaptic mechanisms appear to be mainly responsible for developmental changes in DSI, although presynaptic mechanisms cannot be ruled out entirely. We have also obtained evidence that tonic endocannabinoid release suppresses GABAergic transmission in the mature but not the neonatal hippocampus. The differential DSI magnitude at different stages of maturation could alter synaptic plasticity and learning and memory during hippocampal development. PMID- 20007502 TI - Theta oscillations in primate prefrontal and anterior cingulate cortices in forewarned reaction time tasks. AB - Previously, we introduced a monkey model for human frontal midline theta oscillations as a possible neural correlate of attention. It was based on homologous theta oscillations found in the monkey's prefrontal and anterior cingulate cortices (areas 9 and 32) in a self-initiated hand-movement task. However, it has not been confirmed whether theta activity in the monkey model consistently appears in other situations demanding attention. Here, we examined the detailed properties of theta oscillations in four variations of forewarned reaction time tasks with warning (S1) and imperative (S2) stimuli. We characterized the theta oscillations generated exclusively in areas 9 and 32, as follows: 1) in the S1-S2 interval where movement preparation and reward expectation were presumably involved, the theta power was higher than in the pre S1 period; 2) in the no-go trials of go/no-go tasks instructed by S1, the theta power in the S1-S2 interval was lower than in the pre-S1 period in an asymmetrical reward condition, whereas it was moderately higher in a symmetrical condition; 3) the theta power after reward delivery was higher than in the unrewarded trials; 4) the theta power in the pre-S1 period was higher than in the resting condition; and 5) when the monkey had to guess the S1-S2 duration internally without seeing S2, the theta power in the pre-S1 period was higher than in the original S1-S2 experiment. These findings suggest that attentional loads associated with different causes can induce the same theta activity, thereby supporting the consistency of attention-dependent theta oscillations in our model. PMID- 20007501 TI - Merging of healthy motor modules predicts reduced locomotor performance and muscle coordination complexity post-stroke. AB - Evidence suggests that the nervous system controls motor tasks using a low dimensional modular organization of muscle activation. However, it is not clear if such an organization applies to coordination of human walking, nor how nervous system injury may alter the organization of motor modules and their biomechanical outputs. We first tested the hypothesis that muscle activation patterns during walking are produced through the variable activation of a small set of motor modules. In 20 healthy control subjects, EMG signals from eight leg muscles were measured across a range of walking speeds. Four motor modules identified through nonnegative matrix factorization were sufficient to account for variability of muscle activation from step to step and across speeds. Next, consistent with the clinical notion of abnormal limb flexion-extension synergies post-stroke, we tested the hypothesis that subjects with post-stroke hemiparesis would have altered motor modules, leading to impaired walking performance. In post-stroke subjects (n = 55), a less complex coordination pattern was shown. Fewer modules were needed to account for muscle activation during walking at preferred speed compared with controls. Fewer modules resulted from merging of the modules observed in healthy controls, suggesting reduced independence of neural control signals. The number of modules was correlated to preferred walking speed, speed modulation, step length asymmetry, and propulsive asymmetry. Our results suggest a common modular organization of muscle coordination underlying walking in both healthy and post-stroke subjects. Identification of motor modules may lead to new insight into impaired locomotor coordination and the underlying neural systems. PMID- 20007503 TI - Representation of Horizontal head-on-body position in the primate superior colliculus. AB - Movement-related activity within the superior colliculus (SC) represents the desired displacement of an impending gaze shift. This representation must ultimately be transformed into position-based reference frames appropriate for coordinated eye-head gaze shifts. Parietal areas that project to the SC are modulated by the initial position of both the eye-re-head and head-re-body and SC activity is modulated by eye-re-head position. These considerations led us to investigate whether SC activity is modulated by the head-re-body position. We recorded activity from movement-related SC neurons while head-restrained monkeys performed a delayed-saccade task. Across blocks of trials, the horizontal position of the body was rotated under a space-fixed head to three to five different positions spanning +/-25 degrees . We observed a significant influence of body-under-head position on SC activity in 50/60 neurons. This influence was expressed predominantly as a linear gain field, scaling task-related SC activity without changing the location of the response field (linear gain fields explained >/=20% of the variance in neural activity in approximately 50% of our sample). Smaller nonlinear modulations were also observed in roughly 30% of our sample. SC activity was equally likely to increase or decrease as the body was rotated to the side of neuronal recording and we found no systematic relationship between the directionality or magnitude of the linear gain field with recording location in the SC. We conclude that a signal conveying head-re-body position is present in the SC. Although the functional significance remains open, our findings are consistent with the SC contributing to a displacement-to-position transformation for oculomotor control. PMID- 20007504 TI - Comprehensive follow-up of the first genome-wide association study of multiple sclerosis identifies KIF21B and TMEM39A as susceptibility loci. AB - Genome-wide association studies (GWASs) have proven highly effective, identifying hundreds of associations across numerous complex diseases. These studies typically test hundreds of thousands of variations and identify hundreds of potential associations. However, to date, follow-up attempts have generally only concentrated on just the few most significant initial associations, leaving the majority of true associations in any GWAS study without replication. Here, we present a substantially more comprehensive follow-up of the first genome-wide association screen performed in multiple sclerosis (MS), a complex genetic disease with central nervous system inflammation. We genotyped approximately 30 000 single-nucleotide polymorphisms (SNPs) that demonstrated mild-to-moderate levels of significance (P < or = 0.10) in the initial GWAS in an independent set of 1343 MS cases and 1379 controls. We further replicated several of the most significant findings in another independent data set of 2164 MS cases and 2016 controls. We find considerable evidence for a number of novel susceptibility loci including KIF21B [rs12122721, combined P = 6.56 x 10(-10), odds ratio (OR) = 1.22] and TMEM39A (rs1132200, P = 3.09 x 10(-8), OR = 1.24), both of which meet genome-wide significance. Both of these loci were overlooked in the initial replication, despite being among the top 3000 ( approximately 1%) SNP hits in the original screen. PMID- 20007505 TI - Analysis of the IGF2/H19 imprinting control region uncovers new genetic defects, including mutations of OCT-binding sequences, in patients with 11p15 fetal growth disorders. AB - The imprinted expression of the IGF2 and H19 genes is controlled by the imprinting control region 1 (ICR1) located at chromosome 11p15.5. This methylation-sensitive chromatin insulator works by binding the zinc-finger protein CTCF in a parent-specific manner. DNA methylation defects involving the ICR1 H19/IGF2 domain result in two growth disorders with opposite phenotypes: an overgrowth disorder, the Beckwith-Wiedemann syndrome (maternal ICR1 gain of methylation in 10% of BWS cases) and a growth retardation disorder, the Silver Russell syndrome (paternal ICR1 loss of methylation in 60% of SRS cases). Although a few deletions removing part of ICR1 have been described in some familial BWS cases, little information is available regarding the mechanism of ICR1 DNA methylation defects. We investigated the CTCF gene and the ICR1 domain in 21 BWS patients with ICR1 gain of methylation and 16 SRS patients with ICR1 loss of methylation. We identified four constitutional ICR1 genetic defects in BWS patients, including a familial case. Three of those defects are newly identified imprinting defects consisting of small deletions and a single mutation, which do not involve one of the CTCF binding sites. Moreover, two of those defects affect OCT-binding sequences which are suggested to maintain the unmethylated state of the maternal allele. A single-nucleotide variation was identified in a SRS patient. Our data extends the spectrum of constitutive genetic ICR1 abnormalities and suggests that extensive and accurate analysis of ICR1 is required for appropriate genetic counseling in BWS patients with ICR1 gain of methylation. PMID- 20007506 TI - Quantitative methylation analysis of developmentally important genes in human pregnancy losses after ART and spontaneous conception. AB - To study possible effects of assisted reproductive technologies (ART) on epigenetic reprogramming, we have analyzed the DNA methylation levels of differentially methylated regions (DMRs) of seven imprinted genes (H19, MEG3, LIT1, MEST, NESP55, PEG3 and SNRPN) as well as the promoter regions of the pluripotency gene NANOG and the tumor suppressor gene APC in chorionic villus samples (CVS) of 42 spontaneous miscarriages and stillbirths after ART and 29 abortions/stillbirths after spontaneous conception. We did not find an increased rate of faulty methylation patterns after ART, but significant and trend differences (ROC curve analysis, Wilcoxon test) in the methylation levels of LIT1 (P = 0.006) and H19 (P = 0.085) between ART and non-ART samples. With the possible exception of NANOG, we did not observe a gestational age effect on the methylation levels of the studied genes. The frequency of extreme methylation values in PEG3 and APC was markedly higher than in the other studied genes, indicating an increased susceptibility of some genes to epigenetic alterations. Most methylation abnormalities in CVS represented either hypermethylated DMRs of paternally and maternally imprinted genes or hypomethylated promoters of non imprinted genes. The observed methylation abnormalities (mosaicism) are consistent with methylation reprogramming defects during early embryogenesis. PMID- 20007508 TI - A generalized model of gene dosage and dominant negative effects in macromolecular complexes. AB - Most genes and their corresponding products are supposed to be involved in genetic or biochemical interactions. A typical example is provided by macromolecular complexes, which may contain tens of proteins in defined stoichiometric proportions. Stoichiometric imbalances in such complexes can be a source of abnormal phenotypes. Comparable effects can also arise from negative dominance/transdominance, even though the underlying mechanisms are different. Here I propose a general yet simple biochemical model accounting for the effects of dosage changes and weak dominant/transdominant negative mutations in macromolecular complexes. The molecular alterations studied are predicted to lead to synergistic effects that can drive total multimer concentration and/or activity in a multiple heterozygote below a critical threshold required to ensure a normal phenotype, thus providing an explanation for the phenomenon of unlinked noncomplementation or nonallelic noncomplementation. The model also helps in understanding the basis of heterosis and the long-term consequences of gene dosage alterations and weak dominant/transdominant negative effects. Indeed, it can explain the observed extensive retention of paralogs in polyploids. Finally, because the effects of weak single-gene alterations can escape selection, they may accumulate in the population. This situation has important evolutionary consequences and may eventually lead to reproductive isolation and speciation. Veitia, R. A. A generalized model of gene dosage and dominant negative effects in macromolecular complexes. PMID- 20007507 TI - A novel human heparanase splice variant, T5, endowed with protumorigenic characteristics. AB - Heparanase is a mammalian endo-beta-d-glucuronidase that can cleave heparan sulfate side chains, an activity strongly implicated in tumor cell dissemination. The current study aimed to identify and characterize heparanase splice variants. LEADS, Compugen's alternative splicing modeling platform (Compugen, Tel Aviv, Israel), was used to search for splice variants in silico; tumor-derived cell lines (i.e., CAG myeloma) and tumor biopsies were utilized to validate T5 expression in vivo; signaling (i.e., Src phosphorylation) was evaluated following T5 gene silencing or overexpression and correlated with cell proliferation, colony formation, and tumor xenograft development. A novel spliced form of human heparanase, termed T5, was identified. In this splice variant, 144 bp of intron 5 are joined with exon 4, which results in a truncated, enzymatically inactive protein. T5 overexpression resulted in increased cell proliferation and larger colonies in soft agar, mediated by Src activation. Furthermore, T5 overexpression markedly enhanced tumor xenograft development. T5 expression is up-regulated in 75% of human renal cell carcinoma biopsies examined, which suggests that this splice variant is clinically relevant. Controls included cells overexpressing wild-type heparanase or an empty plasmid and normal-looking tissue adjacent the carcinoma lesion. T5 is a novel functional splice variant of human heparanase endowed with protumorigenic characteristics.-Barash, U., Cohen-Kaplan, V., Arvatz, G., Gingis-Velitski, S., Levy-Adam, F., Nativ, O., Shemesh, R., Ayalon Sofer, M., Ilan, N., Vlodavsky, I. A novel human heparanase splice variant, T5, endowed with protumorigenic characteristics. PMID- 20007510 TI - The crystal structures of two cuticle-degrading proteases from nematophagous fungi and their contribution to infection against nematodes. AB - Cuticle-degrading proteases are involved in the breakdown of cuticle/eggshells of nematodes or insects, a hard physical barrier against fungal infections. Understanding the 3-dimensional structures of these proteins can provide crucial information for improving the effectiveness of these fungi in biocontrol applications, e.g., by targeted protein engineering. However, the structures of these proteases remain unknown. Here, we report the structures of two cuticle degrading proteases from two species of nematophagous fungi. The two structures were solved with X-ray crystallography to resolutions of 1.65 A (Ver112) and 2.1 A (PL646), respectively. Crystal structures of PL646 and Ver112 were found to be very similar to each other, and similar to that of proteinase K from another fungus Tritirachium album. Differences between the structures were found among residues of the substrate binding sites (S1 and S4). Experimental studies showed that the enzymes differed in hydrolytic activity to synthetic peptide substrates. Our analyses of the hydrophobic/hydrophilic and electrostatic features of these two proteins suggest that their surfaces likely play important roles during fungal infection against nematodes. The two crystal structures provide a solid basis for investigating the relationship between structure and function of cuticle-degrading proteases. PMID- 20007509 TI - Acid ceramidase improves the quality of oocytes and embryos and the outcome of in vitro fertilization. AB - A major challenge of assisted reproduction technologies (ARTs) is to mimic the natural environment required to sustain oocyte and embryo survival. Herein, we show that the ceramide-metabolizing enzyme, acid ceramidase (AC), is expressed in human cumulus cells and follicular fluid, essential components of this environment, and that the levels of this enzyme are positively correlated with the quality of human embryos formed in vitro. These observations led us to develop a new approach for oocyte and embryo culture that markedly improved the outcome of in vitro fertilization (IVF). The addition of recombinant AC (rAC) to human and mouse oocyte culture medium maintained their healthy morphology in vitro. Following fertilization, the number of mouse embryos formed in the presence of rAC also was improved (from approximately 40 to 88%), leading to approximately 5-fold more healthy births. To confirm these observations, immature bovine oocytes were matured in vitro and subjected to IVF in the presence of rAC. Significantly more high-grade blastocysts were formed, and the number of morphologically intact, hatched embryos was increased from approximately 24 to 70%. Overall, these data identify AC as an important component of the in vivo oocyte and embryo environment, and provide a novel technology for enhancing the outcome of assisted fertilization. Eliyahu, E., Shtraizent, N., Martinuzzi, K., Barritt, J., He, X., Wei, H., Chaubal, S., Copperman, A. B., Schuchman, E. H. Acid ceramidase improves the quality of oocytes and embryos and the outcome of in vitro fertilization. PMID- 20007512 TI - Oxidative stress in carotid body contributes to enhanced chemoreflex in heart failure: focus on "Elevated mitochondrial superoxide contributes to enhanced chemoreflex in heart failure rabbits". PMID- 20007511 TI - 14-3-3zeta escorts CCTalpha for calcium-activated nuclear import in lung epithelia. AB - Integrity of animal biomembranes is critical to preserve normal cellular functions and viability. Phosphatidylcholine, an indispensible membrane component, requires the enzyme CCTalpha for its biosynthesis. Nuclear expression of CCTalpha is needed for expansion of the nuclear membrane network, but mechanisms for CCTalpha nuclear import are unknown. Herein, we show that in epithelia, extracellular Ca(2+) triggers CCTalpha cytoplasmic-nuclear translocation. CCTalpha nuclear import was associated with binding to 14-3-3zeta, a key regulator of protein trafficking. 14-3-3zeta was both sufficient and required for CCTalpha nuclear import. Helix G within the 14-3-3zeta binding groove interacts with a putative molecular signature within the CCTalpha carboxyl terminal phosphoserine motif (residues 328-343). 14-3-3zeta was critically involved in preserving phosphatidylcholine synthesis and cell viability in a model of Pseudomonas aeruginosa infection where Ca(2+) concentrations increase within epithelia. Thus, 14-3-3zeta controls CCTalpha nuclear import in response to calcium signals, thereby regulating mammalian phospholipid synthesis. Agassandian, M., Chen, B. B., Schuster, C. C., Houtman, J. C. D., Mallampalli, R. K. 14-3-3zeta escorts CCTalpha for calcium-activated nuclear import in lung epithelia. PMID- 20007513 TI - GTPase-Rac enhances depolarization-induced superoxide production by the macula densa during tubuloglomerular feedback. AB - Superoxide (O(2)(-) ) enhances tubuloglomerular feedback (TGF) by scavenging nitric oxide at the macula densa (MD). The primary source of O(2)(-) in the MD during TGF is NADPH oxidase, which is activated by membrane depolarization. While Rac, a small GTP-binding protein, has been shown to enhance NADPH oxidase activity, its role in O(2)(-) generation by the MD is unknown. We hypothesized that depolarization of the MD leads to translocation of Rac to the apical membrane, and its activation, in turn, augments O(2)(-) generation during TGF. We tested this by measuring membrane potential and increased O(2)(-) levels during TGF responses in isolated, perfused tubules containing the intact MD plaque. Switching tubular NaCl from 10 to 80 mM, which induces TGF, depolarized membrane potential by 28.4 + or - 4.5% from control (P < 0.05) and O(2)(-) levels from 124 + or - 19 to 361 + or - 27 U/min. This NaCl-induced depolarization and O(2)(-) generation were blocked by a Cl(-) channel blocker, 5-nitro-2(3 phenylpropylamino) benzoic acid (NPPB; 10(-6) M). Inhibition of Rac blunted NaCl induced O(2)(-) generation by 47%. When the NaCl content of the MD perfusate was increased from 10 to 80 mM, immunointensity of Rac on the apical side increased from 32 + or - 3.1 to 46 + or - 2.5% of the total immunofluorescence in the MD, indicating that high NaCl induces the translocation of Rac to the apical membrane. This NaCl-induced Rac translocation was blocked by a Cl(-) channel blocker, NPPB, indicating that depolarization of the MD induced Rac translocation. In conclusion, we found that depolarization of the MD during TGF leads to translocation of Rac to the apical membrane, which enhances O(2)(-) generation by the MD. PMID- 20007514 TI - A central role for the periphery in the rapid action of cocaine on brain neurons: focus on "Rapid EEG desynchronization and EMG activation induced by intravenous cocaine in freely moving rats: a peripheral, nondopamine neural triggering". PMID- 20007515 TI - Nitric oxide synthase inhibition during treadmill exercise reveals fiber-type specific vascular control in the rat hindlimb. AB - The control of vascular tone during exercise is highly complex and integrated. Specifically, in regards to the contribution of nitric oxide (NO), the observed magnitude and muscle fiber-type dependency of the NO contribution to exercise hyperemia may differ depending on the timing of NO synthase (NOS) inhibition with respect to the exercise bout (i.e., administration prior to vs. during exercise). We tested the hypothesis that, in the presence of prior cyclooxygenase inhibition (indomethacin, 5 mg/kg(-1)), NOS inhibition (N(G)-nitro-L-arginine methyl ester, L-NAME; 10 mg/kg) administered during submaximal treadmill exercise would blunt blood flow and vascular conductance (VC) in the hindlimb muscle(s) of the rat with the greatest reductions in blood flow and VC occurring in the predominantly oxidative muscles. Adult female Wistar rats (n = 10, age: 3-4 mo) ran on a motor driven treadmill (20 m/min, 10% grade). Total and regional hindlimb muscle blood flow and VC were determined via radiolabeled microspheres before (control) and after L-NAME administration during exercise. L-NAME reduced (P < 0.05) total hindlimb muscle blood flow (control: 123 + or - 10, L-NAME: 103 + or - 7 ml x min(-1) x 100g(-1)) and VC (control: 0.95 + or - 0.09, L-NAME: 0.63 + or - 0.05 ml x min(-1) x 100g(-1) x mmHg(-1)). There was a significant correlation (r = 0.51, P < 0.05) between the absolute reductions in VC after L-NAME and the percent sum of type I and IIa fibers in the individual muscles and muscle parts; however, there was no correlation (P = 0.62) when expressed as blood flow. Surprisingly, the highly oxidative muscles demonstrated a marked ability to maintain oxygen delivery, which differs substantially from previous reports of L NAME infusion prior to exercise in these muscles. The demonstration that NO is an important regulator of blood flow and VC in the rat hindlimb during treadmill exercise, but that the fiber-type dependency of NO is altered markedly when NOS inhibition is performed during, vs. prior to, exercise, lends important insights into the integrated nature of vascular control during exercise. PMID- 20007516 TI - Antisickling property of fetal hemoglobin enhances nitric oxide bioavailability and ameliorates organ oxidative stress in transgenic-knockout sickle mice. AB - In sickle cell disease (SCD), the events originating from hemoglobin S polymerization and intravascular sickling lead to reperfusion injury, hemolysis, decreased nitric oxide (NO) bioavailability, and oxidative stress. Oxidative stress is implicated as a contributing factor to multiple organ damage in SCD. We hypothesize that inhibition of sickling by genetic manipulation to enhance antisickling fetal hemoglobin (HbF) expression will have an ameliorating effect on oxidative stress by decreasing intravascular sickling and hemolysis and enhancing NO bioavailability. We tested this hypothesis in BERK (Berkeley) mice expressing exclusively human alpha- and beta(S)-globins and varying levels of HbF, i.e., BERK (<1% HbF), BERKgammaM (20% HbF) and BERKgammaH (40% HbF). Intravascular sickling showed a distinct decrease with increased expression of HbF, which was accompanied by decreased hemolysis and increased NO metabolites (NO(x)) levels. Consistent with decreased intravascular sickling and increased NO bioavailability, BERKgammaM and BERKgammaH mice showed markedly decreased lipid peroxidation accompanied by increased activity/levels of antioxidants [superoxide dismutase (SOD), catalase, glutathione peroxidase (GPx), and reduced glutathione (GSH)] in the muscle, kidney, and liver compared with BERK mice (P < 0.05 0.0001). NO(x) levels showed a strong inverse correlation with hemolytic rate and oxidative stress. Decreased oxidative stress in the presence of elevated HbF levels led to an anti-inflammatory effect as evidenced by decreased peripheral leukocyte counts. These results show that the protective effect of HbF is mediated primarily by decreasing intravascular sickling resulting in decreased oxidative stress and increased NO bioavailability. PMID- 20007517 TI - Effects of insulin-like growth factor-I, insulin, and leucine on protein turnover and ubiquitin ligase expression in rainbow trout primary myocytes. AB - The effects of insulin-like growth factor-I (IGF-I), insulin, and leucine on protein turnover and pathways that regulate proteolytic gene expression and protein polyubiquitination were investigated in primary cultures of 4-day-old rainbow trout myocytes. Supplementing media with 100 nM IGF-I increased protein synthesis by 13% (P < 0.05) and decreased protein degradation by 14% (P < 0.05). Treatment with 1 microM insulin increased protein synthesis by 13% (P < 0.05) and decreased protein degradation by 17% (P < 0.05). Supplementing media containing 0.6 mM leucine with an additional 2.5 mM leucine did not increase protein synthesis rates but reduced rates of protein degradation by 8% (P < 0.05). IGF-I (1 nM-100 nM) and insulin (1 nM-1 microM) independently reduced the abundance of ubiquitin ligase mRNA in a dose-dependent manner, with maximal reductions of approximately 70% for muscle atrophy F-box (Fbx) 32, 40% for Fbx25, and 25% for muscle RING finger-1 (MuRF1, P < 0.05). IGF-I and insulin stimulated phosphorylation of FOXO1 and FOXO4 (P < 0.05), which was inhibited by the phosphatidylinositol 3-kinase (PI 3-kinase) inhibitor wortmannin, and decreased the abundance of polyubiquitinated proteins by 10-20% (P < 0.05). Supplementing media with leucine reduced Fbx32 expression by 25% (P < 0.05) but did not affect Fbx25 nor MuRF1 transcript abundance. Serum deprivation decreased rates of protein synthesis by 60% (P < 0.05), increased protein degradation by 40% (P < 0.05), and increased expression of all ubiquitin ligases. These data suggest that, similar to mammals, the inhibitory effects of IGF-I and insulin on proteolysis occur via P I3-kinase/protein kinase B signaling and are partially responsible for the ability of these compounds to promote protein accretion. PMID- 20007518 TI - Eccentric contraction induces inflammatory responses in rat skeletal muscle: role of tumor necrosis factor-alpha. AB - Eccentric contraction (EC) is known to elicit inflammation and damage in skeletal muscle. Proinflammatory cytokine TNF-alpha plays an important role in this pathogenesis, but the time course of its response to EC and the regulatory mechanisms involved are not clear. The purpose of the study is twofold: 1) to investigate the gene expression of TNF-alpha in rat muscle during and after an acute bout of downhill running and the associated oxidoreductive (redox) changes; and 2) to examine whether EC activates muscle ubiquitin-proteolytic pathway resulting in necrosis and oxidative damage. Female Sprague-Dawley rats (age 3 mo) were randomly divided into five groups (n = 6) that ran on treadmill at 25 m/min at -10% grade for 1 h (group 1) or 2 h (group 2) and were killed immediately; ran for 2 h and killed at 6 h after exercise (group 3), ran for 2 h and killed at 24 h after exercise (group 4); and killed at rest as controls (group 5). TNF-alpha mRNA and protein content showed progressive increases in the deep portion of vastus lateralis (DVL) and gastrocnemius muscles during and after EC. These changes were accompanied by a progressive decrease of mitochondrial aconitase activity and NF-kappaB activation. After 2 h of exercise, elevated levels of serum TNF-alpha, endotoxin, creatine kinase, and lipid peroxidation marker were evident and persisted through 24 h postexercise. At 24 h, there were marked increases in H(2)O(2) concentration, myleoperoxidase activity, and endotoxin level, along with nuclear accumulation of p65, in both muscles. mRNA level of ubiquitin-conjugating enzymes (E(2))-14k was progressively upregulated during exercise and recovery, whereas the expression of the Toll-like receptor 4 (TLR4) in DVL was downregulated in both muscles. We conclude that prolonged EC induces TNF-alpha expression possibly due to NF-kappaB activation stimulated by increased reactive oxygen species generation and endotoxin release. These inflammatory and prooxidative responses may underlie the processes of muscle proteolysis and oxidative damage. PMID- 20007521 TI - Impairment of maximal aerobic power with moderate hypoxia in endurance athletes: do skeletal muscle mitochondria play a role? AB - This study investigates the role of central vs. peripheral factors in the limitation of maximal oxygen uptake (Vo(2max)) with moderate hypoxia [inspired fraction (Fi(O(2))) =14.5%]. Fifteen endurance-trained athletes performed maximal cycle incremental tests to assess Vo(2max), maximal cardiac output (Q(max)), and maximal arteriovenous oxygen (a-vO(2)) difference in normoxia and hypoxia. Muscle biopsies of vastus lateralis were taken 1 wk before the cycling tests to evaluate maximal muscle oxidative capacity (V(max)) and sensitivity of mitochondrial respiration to ADP (K(m)) on permeabilized muscle fibers in situ. Those athletes exhibiting the largest reduction of Vo(2max) in moderate hypoxia (Severe Loss group: -18 +/- 2%) suffered from significant reductions in Q(max) (-4 +/- 1%) and maximal a-vO(2) difference (-14 +/- 2%). Athletes who well tolerated hypoxia, as attested by a significantly smaller drop of Vo(2max) with hypoxia (Moderate Loss group: -7 +/- 1%), also display a blunted Q(max) (-9 +/- 2%) but, conversely, were able to maintain maximal a-vO(2) difference (+1 +/- 2%). Though V(max) was similar in the two experimental groups, the smallest reduction of Vo(2max) with moderate hypoxia was observed in those athletes presenting the lowest apparent K(m) for ADP in the presence of creatine (K(m+Cr)). In already-trained athletes with high muscular oxidative capacities, the qualitative, rather than quantitative, aspects of the mitochondrial function may constitute a limiting factor to aerobic ATP turnover when exercising at low Fi(O(2)), presumably through the functional coupling between the mitochondrial creatine kinase and ATP production. This study suggests a potential role for peripheral factors, including the alteration of cellular homeostasis in active muscles, in determining the tolerance to hypoxia in maximally exercising endurance-trained athletes. PMID- 20007519 TI - A peripheral neuroimmune link: glutamate agonists upregulate NMDA NR1 receptor mRNA and protein, vimentin, TNF-alpha, and RANTES in cultured human synoviocytes. AB - Human primary and clonal synovial cells were incubated with glutamate receptor agonists to assess their modulating influence on glutamate receptors N-methyl-d aspartate (NMDA) NR1 and NR2 and inflammatory cytokines to determine potential for paracrine or autocrine (neurocrine) upregulation of glutamate receptors, as has been shown for bone and chondrocytes. Clonal SW982 synoviocytes constitutively express vimentin, smooth muscle actin (SMA), and NMDA NR1 and NR2. Coincubation (6 h) with glutamate agonists NMDA (5 microM), and the NMDA NR1 glycine site activator (+/-)1-aminocyclopentane-cis-1,3-dicarboxylic acid (5 muM), significantly increases cellular mRNA and protein levels of glutamate receptors, as well as increasing vimentin, SMA, tumor necrosis factor-alpha, and RANTES (regulated on activation, normal T-cell expressed and secreted), assessed qualitatively and quantitatively with nucleotide amplification, image analysis of immunocytochemical staining, fluorescein-activated cell sorting, Western blotting, and immunoassays. Human primary synovial cells harvested from patients with arthritic conditions also constitutively expressed NMDA NR1 with increases after agonist treatment. Glutamate receptor agonist-induced increases were blocked by the noncompetitive glutamate antagonist MK-801 (8 microg/ml) and NR1 blocking antibody. Coincubation with glutamate agonists and phorbol 12-myristate 13-acetate, a protein kinase C activator, significantly enhanced mean levels of TNF-alpha and RANTES in SW982 cell supernatants compared with incubation with either agent alone. Increases were diminished with protein kinase inhibitor and NR1 blocking antibody. The functional activation of glutamate receptors on human synoviocytes establishes a neurogenic cell signaling link between neurotransmitter glutamate released from nerve terminals and target cells in the joint capsule. The influence of glutamate on subsequent release of cellular proinflammatory mediators in non-neural tissue for activation of downstream immune events supports a peripheral neuroimmune link in arthritis. PMID- 20007520 TI - Mitochondrial content and distribution changes specific to mouse diaphragm after chronic normobaric hypoxia. AB - Chronic hypoxia reduces aerobic capacity (mitochondrial content) in limb skeletal muscles, and one of the causes seems to be decreased physical activity. Diaphragm and other respiratory muscles, however, may have a different pattern of adaptation as hypoxia increases the work of breathing. Thus, we hypothesized that chronic hypoxia would not reduce mitochondrial content in mouse diaphragm. Adult male C57BL/6J mice were kept in normoxia (Fi(O(2)) = 21%, control) or normobaric hypoxia (Fi(O(2)) = 10%, hypoxia) for 1, 2, and 4 wk. Mice were then killed, and the diaphragm and gastrocnemius muscles collected for analysis. In the diaphragm, cytochrome c oxidase histochemistry showed less intense staining in the hypoxia group. The total content of subunits from the electron transport chain, pyruvate dehydrogenase kinase 1 (PDK1), and voltage-dependent anion channel 1 (VDAC1) was evaluated by Western blot. These proteins decreased by 25-30% after 4 wk of hypoxia (P < 0.05 vs. control for all comparisons), matching a comparable decrease in diaphragmatic mitochondrial volume density (control 33.6 +/- 5.5% vs. hypoxia 26.8 +/- 6.7%, P = 0.013). Mitochondrial volume density or protein content did not change in gastrocnemius after hypoxia. Hypoxia decreased the content of peroxisome proliferator-activated receptor gamma (PPARgamma) and PPARgamma cofactor 1-alpha (PGC-1alpha) in diaphragm but not in gastrocnemius. PGC-1alpha mRNA levels in diaphragm were also reduced with hypoxia. BCL2/adenovirus E1B interacting protein 3 (BNIP-3) mRNA levels were upregulated after 1 and 2 wk of hypoxia in diaphragm and gastrocnemius, respectively; BNIP-3 protein content increased only in the diaphragm after 4 wk of hypoxia. Contrary to our hypothesis, these results show that chronic hypoxia decreases mitochondrial content in mouse diaphragm, despite the increase in workload. A combination of reduced mitochondrial biogenesis and increased mitophagy seems to be responsible for the decrease in mitochondrial content in the mouse diaphragm after hypoxia. PMID- 20007522 TI - The antidiuretic neurohormone RhoprCAPA-2 downregulates fluid transport across the anterior midgut in the blood-feeding insect Rhodnius prolixus. AB - Osmotic balance in insects is regulated by the excretory system, consisting of Malpighian tubules and the gut under the control of diuretic and antidiuretic factors. Terrestrial insects must conserve water, and antidiuresis is the norm, only interrupted by brief diuretic periods. Surprisingly, little is known about antidiuresis in insects. Two antidiuretic strategies have been described. The first antidiuretic mechanism involves the reabsorption of fluid from the primary urine in the hindgut. More recently, a second antidiuretic strategy was reported, consisting of inhibition of primary urine formation by the Malpighian tubules. Recently, we isolated, characterized, and cloned the gene encoding for the antidiuretic neurohormone (the neuropeptide RhoprCAPA-2) acting on the Malpighian tubules of Rhodnius prolixus. Here we describe a third, novel mechanism central to the antidiuretic strategy of R. prolixus, the inhibition of ion and fluid transport across the anterior midgut by RhoprCAPA-2. Our results show that RhoprCAPA-2 (1 micromol/l) reduces serotonin-stimulated fluid transport from 83 +/- 11 to 12 +/- 12 nl/min and equivalent short-circuit current from 20 +/- 4 to 5 +/- 0.7 microA/cm(2) in diuretic hormone-stimulated anterior midgut. RhoprCAPA 2 appears to function independently of intracellular cGMP or Ca(2+) in the midgut. Thus, the antidiuretic neurohormone RhoprCAPA-2 has multiple target tissues, and we hypothesize that RhoprCAPA-2 functions to coordinate the transport activity of the anterior midgut and Malpighian tubules so that the rate of fluid transport into the haemolymph by the anterior midgut matches the transport rate of Malpighian tubules to maintain the volume and ion composition of haemolymph. PMID- 20007523 TI - Early intervention in Alzheimer disease: The importance of APOE4 plus family history. PMID- 20007524 TI - Association of parental dementia with cognitive and brain MRI measures in middle aged adults. AB - OBJECTIVES: Studies of autosomal dominant Alzheimer disease (AD) have shown structural and cognitive changes in mutation carriers decades prior to clinical disease. Whether such changes are detectable in offspring of persons with sporadic dementia remains unknown. We related prospectively verified parental dementia to brain MRI and cognitive testing in the offspring, within a 2 generational community-based cohort. METHODS: A total of 717 Framingham offspring (mean age: 59 +/- 8 years) were studied. In multivariate analyses, we compared offspring with and without verified parental dementia (and AD) for 1) performance on tests of memory, abstract reasoning, and cognitive flexibility, and 2) volumetric brain MRI measures of total cerebral brain volume (TCBV), hippocampal volume (HV), and white matter hyperintensity volume (WMHV), assessed cross sectionally and longitudinally. RESULTS: When testing the association of parental dementia and AD with baseline cognitive performance, we observed an interaction of parental dementia and AD with APOE epsilon4 status (p < 0.002). In APOE epsilon4 carriers only (n = 165), parental dementia was associated with poorer scores on tests of verbal memory (beta = -1.81 +/- 0.53, p < 0.001) and visuospatial memory (beta = -1.73 +/- 0.47, p < 0.001). These associations were stronger for parental AD (beta = -1.97 +/- 0.52, p < 0.001, beta = -1.95 +/- 0.48, p < 0.001), equivalent to 14-16 years of brain aging. Among APOE epsilon4 carriers, offspring of participants with dementia were also more likely to show an annual decline in TCBV in the top quartile (odds ratio = 4.67 [1.26-17.30], p = 0.02). Regardless of APOE epsilon4 status, participants with parental dementia were more likely to be in the highest quartile of decline in executive function test scores (odds ratio = 1.61 [1.02-2.53], p = 0.04). CONCLUSIONS: Among middle aged carriers of the APOE epsilon4 allele, parental dementia and Alzheimer disease were associated with poorer verbal and visuospatial memory and a higher rate of global brain atrophy. PMID- 20007525 TI - Dermatologic infections in cancer patients treated with epidermal growth factor receptor inhibitor therapy. AB - BACKGROUND: Patients treated with epidermal growth factor receptor inhibitors (EGFRIs) frequently experience dermatologic toxic effects. Whereas the impact of these effects on quality of life and EGFRI dosing has been described, their impact on physical health has not been ascertained. We examined the prevalence of infections that complicate dermatologic toxic effects of EGFRIs. METHODS: We used retrospective chart review methods to analyze 221 patients who were treated in the Skin and Eye Reactions to Inhibitors of EGFR and Kinases clinic, a referral clinic for dermatologic toxic effects of cancer therapies. We reviewed results of bacterial cultures, histopathologic assessment of biopsy samples, and immunohistochemical staining of skin specimens for viral pathogens that were recorded in the patients' medical records. Associations between patient demographic and treatment characteristics and the development of infections were examined using the Fisher exact test. All statistical tests were two-sided. RESULTS: Eighty-four (38%) of the 221 patients showed evidence of infection at sites of dermatologic toxic effect. Fifty (22.6%) of the 221 patients had cultures positive for Staphylococcus aureus, and 12 (5.4%) of the 221 patients cultured positive for methicillin-resistant S aureus. Less frequent infections included herpes simplex (3.2%), herpes zoster (1.8%), and dermatophytes (10.4%). The seborrheic region was the most prevalent site of infection, and patients with leukopenia had higher risk for infection than patients who did not have leukopenia (P = .005). Demographic factors and associated treatments were not associated with the occurrence of a dermatologic infection (P > or = .05). CONCLUSIONS: Patients with dermatologic toxic effects following treatment with EGFRIs have a high prevalence of cutaneous infections. Most notably, bacterial infections developed at sites previously affected by dermatologic toxic effects, with leukopenic patients being at greater risk. PMID- 20007526 TI - Re: False-positive results in cancer epidemiology: a plea for epistemological modesty. PMID- 20007527 TI - Crucial role of phospholipase Cepsilon in induction of local skin inflammatory reactions in the elicitation stage of allergic contact hypersensitivity. AB - Phospholipase Cepsilon (PLCepsilon) is an effector of Ras/Rap small GTPases. We previously demonstrated that PLCepsilon plays a crucial role in development of phorbor ester-induced skin inflammation, which is intimately involved in the promotion of skin carcinogenesis. In this study, we have examined its role in local skin inflammatory reactions during development of contact hypersensitivity toward a hapten 2,4-dinitrofluorobenzene (DNFB). PLCepsilon(+/+) and PLCepsilon( /-) mice were sensitized with DNFB, followed by a DNFB challenge on the ears. PLCepsilon(-/-) mice exhibited substantially attenuated inflammatory reactions compared with PLCepsilon(+/+) mice as shown by suppression of ear swelling, neutrophil infiltration, and proinflammatory cytokine production. In contrast, the extent and kinetics of CD4+ T cell infiltration showed no difference depending on the PLCepsilon background. Adoptive transfer of CD4+ T cells from the sensitized mice to naive mice between PLCepsilon(+/+) and PLCepsilon(-/-) backgrounds indicated that PLCepsilon exerts its function in cells other than CD4+ T cells, presumably fibroblasts or keratinocytes of the skin, to augment inflammatory reactions during the elicitation stage of contact hypersensitivity. Moreover, dermal fibroblasts and epidermal keratinocytes cultured from the skin expressed proinflammatory cytokines in a PLCepsilon-dependent manner on stimulation with T cell-derived cytokines such as IL-17, IFN-gamma, TNF-alpha, and IL-4. These results indicate that PLCepsilon plays a crucial role in induction of proinflammatory cytokine expression in fibroblasts and keratinocytes at the challenged sites, where infiltrated CD4+ T cells produce their intrinsic cytokines, thereby augmenting the local inflammatory reactions. PMID- 20007528 TI - Pronounced phenotype in activated regulatory T cells during a chronic helminth infection. AB - Although several markers have been associated with the characterization of regulatory T cells (Tregs) and their function, no studies have investigated the dynamics of their phenotype during infection. Since the necessity of Tregs to control immunopathology has been demonstrated, we used the chronic helminth infection model Schistosoma mansoni to address the impact on the Treg gene repertoire. Before gene expression profiling, we first studied the localization and Ag-specific suppressive nature of classically defined Tregs during infection. The presence of Foxp3+ cells was predominantly found in the periphery of granulomas and isolated CD4+CD25(hi)Foxp3+ Tregs from infected mice and blocked IFN-gamma and IL-10 cytokine secretion from infected CD4+CD25- effector T cells. Furthermore, the gene expression patterns of Tregs and effector T cells showed that 474 genes were significantly regulated during schistosomiasis. After k-means clustering, we identified genes exclusively regulated in all four populations, including Foxp3, CD103, GITR, OX40, and CTLA-4--classic Treg markers. During infection, however, several nonclassical genes were upregulated solely within the Treg population, such as Slpi, Gzmb, Mt1, Fabp5, Nfil3, Socs2, Gpr177, and Klrg1. Using RT-PCR, we confirmed aspects of the microarray data and also showed that the expression profile of Tregs from S. mansoni-infected mice is simultaneously unique and comparable with Tregs derived from other infections. PMID- 20007529 TI - Differential responses to Smith D autoantigen by mice with HLA-DR and HLA-DQ transgenes: dominant responses by HLA-DR3 transgenic mice with diversification of autoantibodies to small nuclear ribonucleoprotein, double-stranded DNA, and nuclear antigens. AB - Anti-Smith (Sm) D autoantibodies are specific for systemic lupus erythematosus. In this investigation, the influence of HLA-D genes on immune responses to SmD was investigated. Mice with HLA-DR3, HLA-DR4, HLA-DQ0601, HLA-DQ0604, or HLA-DQ8 transgenes were immunized with recombinant SmD1, and their Ab responses were analyzed. Analysis by ELISA showed that all strains responded well to SmD. However, when synthetic SmD peptides were used as substrate, DR3 mice had the highest Ab response followed by DQ8, DQ0604, DQ0601, and DR4. A similar trend was observed in Western blot analysis using WEHI 7.1 cell lysate as the substrate, with the exception that DR4 mice did not generate detectable amounts of Abs. Only sera from DR3 and DQ0604 mice immunoprecipitated A-ribonucleoprotein (RNP), SmB, and SmD. Intermolecular epitope spreading to A-RNP and SmB was evident in DR3 and DQ0604 mice, as sera depleted of anti-SmD Abs were reactive with these proteins. DR3 mice also generated an immune response to C-RNP. Anti-nuclear Abs were detected in the majority of the DR3 mice, whereas moderate reactivities were seen in DQ0604 and DQ8 mice. Interestingly, only DR3 mice mounted an anti-dsDNA Ab response. Approximately half of the anti-dsDNA Abs were cross-reactive with SmD. Ab responses correlated with the strength of the T cell responses. Thus, HLA-DR3 appears to be the dominant HLA-D gene that determines the magnitude and quality of the anti-SmD immune response. In addition, our findings provide insights into the origin of the anti-dsDNA Abs often detected in patients with systemic lupus erythematosus. PMID- 20007531 TI - Elevated mitochondrial reactive oxygen species generation affects the immune response via hypoxia-inducible factor-1alpha in long-lived Mclk1+/- mouse mutants. AB - Mitochondrial reactive oxygen species (ROS) are believed to stabilize hypoxia inducible factor (HIF)-1alpha, a transcriptional regulator of the immune response. Mclk1 encodes a mitochondrial protein that is necessary for ubiquinone biosynthesis. Heterozygote Mclk1(+/-) mutant mice are long-lived despite increased mitochondrial ROS and decreased energy metabolism. In this study, Mclk1(+/-) mutant mice in the C57BL/6J background displayed increased basal and induced expression of HIF-1alpha in liver and macrophages in association with elevated expression of inflammatory cytokines, in particular TNF-alpha. Mutant macrophages showed increased classical and decreased alternative activation, and mutant mice were hypersensitive to LPS. Consistent with these observations in vivo, knock-down of Mclk1 in murine RAW264.7 macrophage-like cells induced increased mitochondrial ROS as well as elevated expression of HIF-1alpha and secretion of TNF-alpha. We used an antioxidant peptide targeted to mitochondria to show that altered ROS metabolism is necessary for the enhanced expression of HIF-1alpha, which, in turn, is necessary for increased TNF-alpha secretion. These findings provide in vivo evidence for the action of mitochondrial ROS on HIF 1alpha activity and demonstrate that changes in mitochondrial function within physiologically tolerable limits modulate the immune response. Our results further suggest that altered immune function through a limited increase in HIF 1alpha expression can positively impact animal longevity. PMID- 20007530 TI - Mammalian target of rapamycin inhibition and alloantigen-specific regulatory T cells synergize to promote long-term graft survival in immunocompetent recipients. AB - Minimization of immunosuppression and donor-specific tolerance to MHC-mismatched organ grafts are important clinical goals. The therapeutic potential of regulatory T cells (Tregs) has been demonstrated, but conditions for optimizing their in vivo function posttransplant in nonlymphocyte-depleted hosts remain undefined. In this study, we address mechanisms through which inhibition of the mammalian target of rapamycin (Rapa) synergizes with alloantigen-specific Treg (AAsTreg) to permit long-term, donor-specific heart graft survival in immunocompetent hosts. Crucially, immature allogeneic dendritic cells allowed AAsTreg selection in vitro, with minimal expansion of unwanted (Th17) cells. The rendered Treg potently inhibited T cell proliferation in an Ag-specific manner. However, these AAsTreg remained unable to control T cells stimulated by allogeneic mature dendritic cells, a phenomenon dependent on the release of proinflammatory cytokines. In vivo, Rapa administration reduced danger-associated IL-6 production, T cell proliferation, and graft infiltration. Based on these observations, AAsTreg were administered posttransplant (day 7) in combination with a short course of Rapa and rendered >80% long-term (>150 d) graft survival, a result superior to that achieved with polyclonal Treg. Moreover, graft protection was alloantigen-specific. Significantly, long-term graft survival was associated with alloreactive T cell anergy. These findings delineate combination of transient mammalian target of Rapa inhibition with appropriate AAsTreg selection as an effective approach to promote long-term organ graft survival. PMID- 20007532 TI - Surfactant protein B propeptide contains a saposin-like protein domain with antimicrobial activity at low pH. AB - Surfactant protein B (SP-B) proprotein contains three saposin-like protein (SAPLIP) domains: a SAPLIP domain corresponding to the mature SP-B peptide is essential for lung function and postnatal survival; the function of SAPLIP domains in the N-terminal (SP-BN) and C-terminal regions of the proprotein is not known. In the current study, SP-BN was detected in the supernatant of mouse bronchoalveolar lavage fluid (BALF) and in nonciliated bronchiolar cells, alveolar type II epithelial cells, and alveolar macrophages. rSP-BN indirectly promoted the uptake of bacteria by macrophage cell lines and directly killed bacteria at acidic pH, consistent with a lysosomal, antimicrobial function. Native SP-BN isolated from BALF also killed bacteria but only at acidic pH; the bactericidal activity of BALF at acidic pH was completely blocked by SP-BN Ab. Transgenic mice overexpressing SP-BN and mature SP-B peptide had significantly decreased bacterial burden and increased survival following intranasal inoculation with bacteria. These findings support the hypothesis that SP-BN contributes to innate host defense of the lung by supplementing the nonoxidant antimicrobial defenses of alveolar macrophages. PMID- 20007533 TI - Peptide-MHC class II complex stability governs CD4 T cell clonal selection. AB - The clonal composition of the T cell response can affect its ability to mediate infection control or to induce autoimmunity, but the mechanisms regulating the responding TCR repertoire remain poorly defined. In this study, we immunized mice with wild-type or mutated peptides displaying varying binding half-lives with MHC class II molecules to measure the impact of peptide-MHC class II stability on the clonal composition of the CD4 T cell response. We found that, although all peptides elicited similar T cell response size on immunization, the clonotypic diversity of the CD4 T cell response correlated directly with the half-life of the immunizing peptide. Peptides with short half-lives focused CD4 T cell response toward high-affinity clonotypes expressing restricted public TCR, whereas peptides with longer half-lives broadened CD4 T cell response by recruiting lower-affinity clonotypes expressing more diverse TCR. Peptides with longer half-lives did not cause the elimination of high-affinity clonotypes, and at a low dose, they also skewed CD4 T cell response toward higher-affinity clonotypes. Taken collectively, our results suggest the half-life of peptide-MHC class II complexes is the primary parameter that dictates the clonotypic diversity of the responding CD4 T cell compartment. PMID- 20007535 TI - RasGRP1 regulates antigen-induced developmental programming by naive CD8 T cells. AB - Ag encounter by naive CD8 T cells initiates a developmental program consisting of cellular proliferation, changes in gene expression, and the formation of effector and memory T cells. The strength and duration of TCR signaling are known to be important parameters regulating the differentiation of naive CD8 T cells, although the molecular signals arbitrating these processes remain poorly defined. The Ras-guanyl nucleotide exchange factor RasGRP1 has been shown to transduce TCR mediated signals critically required for the maturation of developing thymocytes. To elucidate the role of RasGRP1 in CD8 T cell differentiation, in vitro and in vivo experiments were performed with 2C TCR transgenic CD8 T cells lacking RasGRP1. In this study, we report that RasGRP1 regulates the threshold of T cell activation and Ag-induced expansion, at least in part, through the regulation of IL-2 production. Moreover, RasGRP1(-/-) 2C CD8 T cells exhibit an anergic phenotype in response to cognate Ag stimulation that is partially reversible upon the addition of exogenous IL-2. By contrast, the capacity of IL-2/IL-2R interactions to mediate Ras activation and CD8 T cell expansion and differentiation appears to be largely RasGRP1-independent. Collectively, our results demonstrate that RasGRP1 plays a selective role in T cell signaling, controlling the initiation and duration of CD8 T cell immune responses. PMID- 20007534 TI - IFN regulatory factor 5 is required for disease development in the FcgammaRIIB-/ Yaa and FcgammaRIIB-/- mouse models of systemic lupus erythematosus. AB - Polymorphisms in the transcription factor IFN regulatory factor 5 (IRF5) are strongly associated in human genetic studies with an increased risk of developing the autoimmune disease systemic lupus erythematosus. However, the biological role of IRF5 in lupus pathogenesis has not previously been tested in an animal model. In this study, we show that IRF5 is absolutely required for disease development in the FcgammaRIIB(-/-)Yaa and FcgammaRIIB(-/-) lupus models. In contrast to IRF5 sufficient FcgammaRIIB(-/-)Yaa mice, IRF5-deficient FcgammaRIIB(-/-)Yaa mice do not develop lupus manifestations and have a phenotype comparable to wild-type mice. Strikingly, full expression of IRF5 is required for the development of autoimmunity, as IRF5 heterozygotes had dramatically reduced disease. One effect of IRF5 is to induce the production of the type I IFN, IFN-alpha, a cytokine implicated in lupus pathogenesis. To address the mechanism by which IRF5 promotes disease, we evaluated FcgammaRIIB(-/-)Yaa mice lacking the type I IFN receptor subunit 1. Unlike the IRF5-deficient and IRF5-heterozygous FcgammaRIIB(-/-)Yaa mice, type I IFN receptor subunit 1-deficient FcgammaRIIB(-/-)Yaa mice maintained a substantial level of residual disease. Furthermore, in FcgammaRIIB(-/-) mice lacking Yaa, IRF5-deficiency also markedly reduced disease manifestations, indicating that the beneficial effects of IRF5 deficiency in FcgammaRIIB(-/-)Yaa mice are not due only to inhibition of the enhanced TLR7 signaling associated with the Yaa mutation. Overall, we demonstrate that IRF5 plays an essential role in lupus pathogenesis in murine models and that this is mediated through pathways beyond that of type I IFN production. PMID- 20007536 TI - Ectopic activation of Mycobacterium tuberculosis-specific CD4+ T cells in lungs of CCR7-/- mice. AB - Initiation of an adaptive cellular immune response depends on intimate interactions with APCs and naive T lymphocytes. We previously reported that activation of naive Mycobacterium tuberculosis-specific CD4+ T cells depends on dendritic cell (DC) transport of live bacteria from the lungs to the mediastinal lymph node (MDLN). Because the migratory paths of DCs are largely governed by the chemokine receptor CCR7, which is expressed on DCs upon maturation by proinflammatory stimuli, we examined the quantitative contribution of CCR7 dependent DC migration in the context of tuberculosis. We found that early trafficking of DCs from the lungs to the MDLN depended on CCR7-mediated signaling, but alternative mechanism(s) are used later in infection. Impaired migration of DCs in CCR7(-/-) mice resulted in delayed dissemination of bacteria to MDLN and spleen and in delayed kinetics of activation of adoptively transferred Ag85B-specific CD4+ T cells. Furthermore, in contrast to control mice, we found that naive Ag85B-specific CD4+ T cells are activated to proliferate in the lungs of CCR7(-/-) mice and, when infected with higher doses of bacteria, resistance to M. tuberculosis infection in CCR7(-/-) mice is compromised compared with wild-type mice. PMID- 20007537 TI - Mycobacterium bovis bacillus Calmette-Guerin killed by extended freeze-drying targets plasmacytoid dendritic cells to regulate lung inflammation. AB - We have previously shown that bacillus Calmette-Guerin (BCG) inactivated by extended freeze-drying (EFD) reduces airway hyperresponsiveness, whereas live and heat-killed BCG fail to do so. However, the cells involved in the protective effect and the signaling and transcriptional networks that could reprogram T cell commitment after EFD BCG treatment remained to be elucidated. We investigated whether EFD BCG targets plasmacytoid dendritic cells (pDCs) potentially involved in the polarization of regulatory T cells (Tregs) and the transcriptional factors that regulate allergic inflammation. OVA-sensitized mice were s.c. injected with EFD, live, or heat-killed BCG. We analyzed after the injection of the various BCG preparations: 1) pDCs recruited in the draining lymph nodes (day 4); 2) transcription factors involved in inflammation and T cell commitment in spleen and lungs after OVA challenge (day 28). Airway hyperresponsiveness and transcription factors were determined after in vivo depletion of pDCs or Tregs in EFD BCG-treated and OVA-challenged mice. EFD BCG reduced inflammation via the recruitment of pDCs polarizing the differentiation of naive CD4+ T lymphocytes into Tregs. In vivo, pDC or Treg depletion at the time of EFD BCG treatment abrogated the protection against inflammation. EFD BCG treatment upregulated Forkhead-winged helix transcription factor (Treg signature) and downregulated GATA-3 and RORgammat (Th2 and Th17 signatures) more efficiently than live and heat-killed BCG. Moreover, only EFD BCG enhanced peroxisome proliferator activated receptor gamma expression and blocked NF-kappaB activation, cyclooxygenase expression, and p38 MAPK phosphorylation. EFD BCG reduced allergic inflammation by recruiting pDCs that promoted Tregs; EFD BCG acted as a peroxisome proliferator-activated receptor gamma agonist and thus could be used in asthma and other inflammatory diseases. PMID- 20007538 TI - An NZW-derived interval on chromosome 7 moderates sialadenitis, but not insulitis in congenic nonobese diabetic mice. AB - Autoimmune lymphocytic infiltration of the salivary glands, termed sialadenitis, is a pathologic feature of Sjogren's syndrome (SjS) that is also prominent in nonobese diabetic (NOD) mice. Genetic factors regulate sialadenitis, and a previous (NOD x NZW)F2 study detected linkage to murine chromosome (Chr) 7. The locus, subsequently annotated as Ssial3, maps to the distal end of Chr7 and overlaps a region associated with type 1 diabetes susceptibility in NOD mice. To examine whether Ssial3 could contribute to both diseases, or was specific for SjS, we generated a congenic mouse strain that harbored an NZW-derived Chr7 interval on the NOD genetic background. This congenic strain exhibited reduced sialadenitis compared with NOD mice and confirmed Ssial3. This reduction, however, did not ameliorate saliva abnormalities associated with SjS-like disease in NOD mice, nor were congenic mice protected against insulitis (lymphocytic infiltration of the pancreatic islets) or diabetes onset. Thus, the Ssial3 locus appears to have a tissue-specific effect for which the NZW allele is unable to prevent other autoimmune traits in the NOD mouse. Anomalous increases for antinuclear Ab production and frequency of marginal-zone B cells were also identified in congenic mice, indicating that the NZW-derived Chr7 interval has a complex effect on the NOD immune system. PMID- 20007540 TI - Antigen specificity determines the pro- or antitumoral nature of CD8+ T cells. AB - Although CD8+ T cells are usually considered antitumoral, several recent studies report that the cells can also promote tumor progression. Using the melanoma cell line B16 as a murine model of pulmonary metastasis, we examined whether the pro- versus antitumoral effects of CD8+ T cells relate to their Ag specificity. Results of the study indicate that although CD8+ T cells specific for tumor Ags promote tumor rejection, CD8+ T cells specific for unrelated Ags promote tumor progression. We found the effect to be partly attributable to CD8+ T cells dampening effective antitumor NK cell responses. Notably, activation of CD8+ T cell responses by an unrelated stimulus, in this case infection with influenza virus, increased the number of pulmonary tumor nodules. These data provide a rationale for previously unexplained data identifying contrasting roles for CD8+ T cells in tumor progression. PMID- 20007539 TI - The anti-inflammatory and proresolving mediator resolvin E1 protects mice from bacterial pneumonia and acute lung injury. AB - Whereas pneumonia is the most common cause of death and disability worldwide, most cases of pneumonia spontaneously resolve. Mechanisms that promote pneumonia resolution remain to be determined. Resolvin E1 (RvE1) is an endogenous mediator that displays proresolving actions in sterile inflammation. In this study, we developed a new model of aspiration pneumonia to evaluate the effect of RvE1 on acute lung injury caused by acid aspiration and subsequent bacterial challenge. Mice received hydrochloric acid into the left lung followed by the enteric pathogen Escherichia coli. I.v. administration of RvE1 (approximately 0.005 mg/kg) prior to acid injury selectively decreased lung neutrophil accumulation by 55% and enhanced clearance of E. coli. RvE1 significantly decreased lung tissue levels of several proinflammatory chemokines and cytokines, including IL-1beta, IL-6, HMGB-1, MIP-1alpha, MIP-1beta, keratinocyte-derived chemokine, and MCP-1, in a manner independent of the anti-inflammatory mediators IL-10 and lipoxin A4. In addition, animals treated with RvE1 had a marked improvement in survival. These findings in experimental aspiration pneumonia have uncovered protective roles for RvE1 in pathogen-mediated inflammation that are both anti-inflammatory for neutrophils and protective for host defense, suggesting that RvE1 represents the first candidate for a novel therapeutic strategy for acute lung injury and pneumonia that harnesses natural resolution mechanisms. PMID- 20007541 TI - Transient Epstein-Barr virus reactivation in CD3 monoclonal antibody-treated patients. AB - Here we report a unique situation in which an early and synchronized Epstein-Barr virus (EBV) reactivation was induced by a 6-day course of treatment with a humanized CD3-specific monoclonal antibody in patients with recent onset of type 1 diabetes. The virologic and immunologic analysis demonstrated that this reactivation was transient, self-limited, and isolated, associated with the rapid advent of an EBV-specific T-cell response. The anti-CD3 antibody administration induced short-lasting immunosuppression and minor yet clear-cut signs of T-cell activation that preceded viral reactivation. Early posttransplant monitoring of renal and islet allograft recipients showed that no comparable phenomenon was observed after the administration of full-dose immunosuppressive therapy. This EBV reactivation remains of no apparent clinical concern over the long term and should not preclude further development of therapeutic anti-CD3 antibodies. This phenomenon may also direct new research avenues to understand the still ill defined nature of stimuli triggering EBV reactivation in vivo. PMID- 20007542 TI - Persons with Quebec platelet disorder have a tandem duplication of PLAU, the urokinase plasminogen activator gene. AB - Quebec platelet disorder (QPD) is an autosomal dominant bleeding disorder linked to a region on chromosome 10 that includes PLAU, the urokinase plasminogen activator gene. QPD increases urokinase plasminogen activator mRNA levels, particularly during megakaryocyte differentiation, without altering expression of flanking genes. Because PLAU sequence changes were excluded as the cause of this bleeding disorder, we investigated whether the QPD mutation involved PLAU copy number variation. All 38 subjects with QPD had a direct tandem duplication of a 78-kb genomic segment that includes PLAU. This mutation was specific to QPD as it was not present in any unaffected family members (n = 114), unrelated French Canadians (n = 221), or other persons tested (n = 90). This new information on the genetic mutation will facilitate diagnostic testing for QPD and studies of its pathogenesis and prevalence. QPD is the first bleeding disorder to be associated with a gene duplication event and a PLAU mutation. PMID- 20007543 TI - Interconnecting molecular pathways in the pathogenesis and drug sensitivity of T cell acute lymphoblastic leukemia. AB - To identify dysregulated pathways in distinct phases of NOTCH1-mediated T-cell leukemogenesis, as well as small-molecule inhibitors that could synergize with or substitute for gamma-secretase inhibitors (GSIs) in T-cell acute lymphoblastic leukemia (T-ALL) therapy, we compared gene expression profiles in a Notch1 induced mouse model of T-ALL with those in human T-ALL. The overall patterns of NOTCH1-mediated gene expression in human and mouse T-ALLs were remarkably similar, as defined early in transformation in the mouse by the regulation of MYC and its target genes and activation of nuclear factor-kappaB and PI3K/AKT pathways. Later events in murine Notch1-mediated leukemogenesis included down regulation of genes encoding tumor suppressors and negative cell cycle regulators. Gene set enrichment analysis and connectivity map algorithm predicted that small-molecule inhibitors, including heat-shock protein 90, histone deacetylase, PI3K/AKT, and proteasome inhibitors, could reverse the gene expression changes induced by NOTCH1. When tested in vitro, histone deacetylase, PI3K and proteasome inhibitors synergized with GSI in suppressing T-ALL cell growth in GSI-sensitive cells. Interestingly, alvespimycin, a potent inhibitor of the heat-shock protein 90 molecular chaperone, markedly inhibited the growth of both GSI-sensitive and -resistant T-ALL cells, suggesting that its loss disrupts signal transduction pathways crucial for the growth and survival of T-ALL cells. PMID- 20007545 TI - The P2X7-nonmuscle myosin membrane complex regulates phagocytosis of nonopsonized particles and bacteria by a pathway attenuated by extracellular ATP. AB - Phagocytosis of nonopsonized bacteria is central to innate immunity, but its regulation is less defined. We show that overexpression of the P2X(7) receptor greatly augments the phagocytosis of nonopsonized beads and heat-killed bacteria by transfected HEK-293 cells, whereas blocking P2X(7) expression by siRNA significantly reduces the phagocytic ability of human monocytic cells. An intact P2X(7)-nonmuscle myosin complex is required for phagocytosis of nonopsonized beads because activation of P2X(7) receptors by adenosine triphosphate (ATP), which dissociates myosin IIA from the P2X(7) complex, inhibits this phagocytic pathway. Fresh human monocytes rapidly phagocytosed live and heat-killed Staphylococcus aureus and Escherichia coli in the absence of serum, but the uptake was reduced by prior incubation with ATP, or P2X(7) monoclonal antibody, or recombinant P2X(7) extracellular domain. Injection of beads or bacteria into the peritoneal cavity of mice resulted in their brisk phagocytosis by macrophages, but injection of ATP before particles markedly decreased this uptake. These data demonstrate a novel pathway of phagocytosis of nonopsonized particles and bacteria, which operate in vivo and require an intact P2X(7) nonmuscle myosin IIA membrane complex. The inhibitory effect of ATP on particle uptake by the macrophage is regulated by the P2X(7) receptor and defines this phagocytic pathway. PMID- 20007544 TI - Cbfb/Runx1 repression-independent blockage of differentiation and accumulation of Csf2rb-expressing cells by Cbfb-MYH11. AB - It is known that CBFB-MYH11, the fusion gene generated by inversion of chromosome 16 in human acute myeloid leukemia, is causative for oncogenic transformation. However, the mechanism by which CBFB-MYH11 initiates leukemogenesis is not clear. Previously published reports showed that CBFB-MYH11 dominantly inhibits RUNX1 and CBFB, and such inhibition has been suggested as the mechanism for leukemogenesis. Here we show that Cbfb-MYH11 caused Cbfb/Runx1 repression-independent defects in both primitive and definitive hematopoiesis. During primitive hematopoiesis, Cbfb MYH11 delayed differentiation characterized by sustained expression of Gata2, Il1rl1, and Csf2rb, a phenotype not found in Cbfb and Runx1 knockout mice. Expression of Cbfb-MYH11 in the bone marrow induced the accumulation of abnormal progenitor-like cells expressing Csf2rb in preleukemic mice. The expression of all 3 genes was detected in most human and murine CBFB-MYH11(+) leukemia samples. Interestingly, Cbfb-MYH11(+) preleukemic progenitors and leukemia-initiating cells did not express Csf2rb, although the majority of leukemia cells in our Cbfb MYH11 knockin mice were Csf2rb(+). Therefore Csf2rb can be used as a negative selection marker to enrich preleukemic progenitor cells and leukemia-initiating cells from Cbfb-MYH11 mice. These results suggest that Cbfb/Runx1 repression independent activities contribute to leukemogenesis by Cbfb-MYH11. PMID- 20007546 TI - Retroviral insertional mutagenesis identifies Zeb2 activation as a novel leukemogenic collaborating event in CALM-AF10 transgenic mice. AB - The t(10;11) translocation results in a CALM-AF10 fusion gene in a subset of leukemia patients. Expression of a CALM-AF10 transgene results in leukemia, with prolonged latency and incomplete penetrance, suggesting that additional events are necessary for leukemic transformation. CALM-AF10 mice infected with the MOL4070LTR retrovirus developed acute leukemia, and ligation-mediated polymerase chain reaction was used to identify retroviral insertions at 19 common insertion sites, including Zeb2, Nf1, Mn1, Evi1, Ift57, Mpl, Plag1, Kras, Erg, Vav1, and Gata1. A total of 26% (11 of 42) of the mice had retroviral integrations near Zeb2, a transcriptional corepressor leading to overexpression of the Zeb2 transcript. A total of 91% (10 of 11) of mice with Zeb2 insertions developed B lineage acute lymphoblastic leukemia, suggesting that Zeb2 activation promotes the transformation of CALM-AF10 hematopoietic precursors toward B-lineage leukemias. More than half of the mice with Zeb2 integrations also had Nf1 integrations, suggesting cooperativity among CALM-AF10, Zeb2, and Ras pathway mutations. We searched for Nras, Kras, and Ptpn11 point mutations in the CALM AF10 leukemic mice. Three mutations were identified, all of which occurred in mice with Zeb2 integrations, consistent with the hypothesis that Zeb2 and Ras pathway activation promotes B-lineage leukemic transformation in concert with CALM-AF10. PMID- 20007548 TI - Trisomy of Erg is required for myeloproliferation in a mouse model of Down syndrome. AB - Down syndrome is characterized by multiple phenotypic manifestations associated with trisomy of chromosome 21. The transient myeloproliferative disorder and acute megakaryocytic leukemia associated with Down syndrome are uniquely associated with mutations in the transcription factor GATA1; however, the identity of trisomic genes on chromosome 21 that predispose to these hematologic disorders remains unknown. Using a loss-of-function allele, we show that specific reduction to functional disomy of the Erg gene corrects the pathologic and hematologic features of myeloproliferation in the Ts(17(16))65Dn mouse model of Down syndrome, including megakaryocytosis and progenitor cell expansion. Our data provide genetic evidence establishing the need for Erg trisomy for myeloproliferation in Ts(17(16))65Dn mice and imply that increased ERG gene dosage may be a key consequence of trisomy 21 that can predispose to malignant hematologic disorders in Down syndrome. PMID- 20007547 TI - EF-hand domains of MCFD2 mediate interactions with both LMAN1 and coagulation factor V or VIII. AB - Combined deficiency of factor V and factor VIII (F5F8D) is a bleeding disorder caused by mutations in either LMAN1 or MCFD2. LMAN1 (ERGIC-53) and MCFD2 form a Ca(2+)-dependent cargo receptor that cycles between the endoplasmic reticulum (ER) and the ER-Golgi intermediate compartment for efficient transport of FV/FVIII from the ER to the Golgi. Here we show that the C-terminal EF-hand domains are both necessary and sufficient for MCFD2 to interact with LMAN1. MCFD2 with a deletion of the entire N-terminal non-EF hand region still retains the LMAN1-binding function. Deletions that disrupt core structure of the EF-hand domains abolish LMAN1 binding. Circular dichroism spectroscopy studies on missense mutations localized to different structural elements of the EF-hand domains suggest that Ca(2+)-induced folding is important for LMAN1 interaction. The EF-hand domains also mediate the interaction with FV and FVIII. However, mutations in MCFD2 that disrupt the tertiary structure and abolish LMAN1 binding still retain the FV/FVIII binding activities, suggesting that this interaction is independent of Ca(2+)-induced folding of the protein. Our results suggest that the EF-hand domains of MCFD2 contain separate binding sites for LMAN1 and FV/FVIII that are essential for cargo receptor formation and cargo loading in the ER. PMID- 20007549 TI - A community cluster of oseltamivir-resistant cases of 2009 H1N1 influenza. PMID- 20007551 TI - Act II--curve bending. PMID- 20007550 TI - Payment reform--the need to harmonize approaches in Medicare and the private sector. PMID- 20007552 TI - Week 1 of the Senate debate--amending the reform bill. PMID- 20007553 TI - Ensuring the fiscal sustainability of health care reform. PMID- 20007554 TI - The need for science in the practice of public health. PMID- 20007556 TI - Attitudes toward domestic violence: a cultural perspective. AB - This study examines the effect of acculturation on the attitudes held by Ethiopian Jews in Israel toward domestic violence (DV). The study findings revealed the following: Ethiopians who immigrated to Israel (n = 31) held more lenient attitudes toward DV than Israeli born Jews (n = 62), which supported the hypothesis that culture influences attitudes toward DV; in addition, Ethiopians born in Israel (n = 29) held attitudes closer to those of Israeli-born Jews who were not from Ethiopian origin, thus supporting the hypothesis that integration into the host country results in changes in DV attitudes. These are important findings due to the extremely high number of DV episodes among immigrant populations in general and Ethiopian Jews living in Israel in particular. This study may provide optimism in that it is probable that the younger generation will prove to be less violent than the first-generation immigrants. Perhaps one conclusion that can be drawn is the importance of expediting the integration process of the second-generation Ethiopian Jews in Israel. PMID- 20007555 TI - Clinical features of the initial cases of 2009 pandemic influenza A (H1N1) virus infection in China. AB - BACKGROUND: The first case of 2009 pandemic influenza A (H1N1) virus infection in China was documented on May 10. Subsequently, persons with suspected cases of infection and contacts of those with suspected infection were tested. Persons in whom infection was confirmed were hospitalized and quarantined, and some of them were closely observed for the purpose of investigating the nature and duration of the disease. METHODS: During May and June 2009, we observed 426 persons infected with the 2009 pandemic influenza A (H1N1) virus who were quarantined in 61 hospitals in 20 provinces. Real-time reverse-transcriptase-polymerase-chain reaction (RT-PCR) testing was used to confirm infection, the clinical features of the disease were closely monitored, and 254 patients were treated with oseltamivir within 48 hours after the onset of disease. RESULTS: The mean age of the 426 patients was 23.4 years, and 53.8% were male. The diagnosis was made at ports of entry (in 32.9% of the patients), during quarantine (20.2%), and in the hospital (46.9%). The median incubation period of the virus was 2 days (range, 1 to 7). The most common symptoms were fever (in 67.4% of the patients) and cough (69.5%). The incidence of diarrhea was 2.8%, and the incidence of nausea and vomiting was 1.9%. Lymphopenia, which was common in both adults (68.1%) and children (92.3%), typically occurred on day 2 (range, 1 to 3) and resolved by day 7 (range, 6 to 9). Hypokalemia was observed in 25.4% of the patients. Duration of fever was typically 3 days (range, 1 to 11). The median length of time during which patients had positive real-time RT-PCR test results was 6 days (range, 1 to 17). Independent risk factors for prolonged real-time RT-PCR positivity included an age of less than 14 years, male sex, and a delay from the onset of symptoms to treatment with oseltamivir of more than 48 hours. CONCLUSIONS: Surveillance of the 2009 H1N1 virus in China shows that the majority of those infected have a mild illness. The typical period during which the virus can be detected with the use of real-time RT-PCR is 6 days (whether or not fever is present). The duration of infection may be shortened if oseltamivir is administered. PMID- 20007557 TI - Barriers to disclosing and reporting violence among women in Pakistan: findings from a national household survey and focus group discussions. AB - Worldwide, many women who experience domestic violence keep their experience secret. Few report to official bodies. In a national survey of abuse against women in Pakistan, we examined factors related to disclosure: women who had experienced physical violence telling someone about it. In focus groups, we explored why women do not report domestic violence. Nearly one third of the 23,430 women interviewed had experienced physical violence. Only 35% of them had told anyone about it, almost always someone within their own family. Several personal and family factors were associated with disclosure. Having discussed the issue and feeling empowered to discuss violence were consistent associations. Of the 7,895 women who had suffered physical violence, only 14 had reported the matter to the police. Female focus groups said women who report violence risk their reputation and bring dishonor to the family; women fear reporting violence because it may exacerbate the problem and may lead to separation or divorce and loss of their children. Focus groups of men and women were skeptical about community leaders, councilors, and religious leaders supporting reporting of violence. They suggested setting up local groups where abused women could seek help and advice. There are strong disincentives to reporting violence in Pakistan, which are well known to women. Until better systems for reporting and dealing with reported cases are in place, domestic violence will continue to be a hidden scourge here and elsewhere. PMID- 20007558 TI - Images in clinical medicine. Ileocecal intussusception. PMID- 20007559 TI - The breadth of hopes. PMID- 20007560 TI - Allogeneic hematopoietic stem-cell transplantation for sickle cell disease. AB - BACKGROUND: Myeloablative allogeneic hematopoietic stem-cell transplantation is curative in children with sickle cell disease, but in adults the procedure is unduly toxic. Graft rejection and graft-versus-host disease (GVHD) are additional barriers to its success. We performed nonmyeloablative stem-cell transplantation in adults with sickle cell disease. METHODS: Ten adults (age range, 16 to 45 years) with severe sickle cell disease underwent nonmyeloablative transplantation with CD34+ peripheral-blood stem cells, mobilized by granulocyte colony stimulating factor (G-CSF), which were obtained from HLA-matched siblings. The patients received 300 cGy of total-body irradiation plus alemtuzumab before transplantation, and sirolimus was administered afterward. RESULTS: All 10 patients were alive at a median follow-up of 30 months after transplantation (range, 15 to 54). Nine patients had long-term, stable donor lymphohematopoietic engraftment at levels that sufficed to reverse the sickle cell disease phenotype. Mean (+/-SE) donor-recipient chimerism for T cells (CD3+) and myeloid cells (CD14+15+) was 53.3+/-8.6% and 83.3+/-10.3%, respectively, in the nine patients whose grafts were successful. Hemoglobin values before transplantation and at the last follow-up assessment were 9.0+/-0.3 and 12.6+/-0.5 g per deciliter, respectively. Serious adverse events included the narcotic-withdrawal syndrome and sirolimus-associated pneumonitis and arthralgia. Neither acute nor chronic GVHD developed in any patient. CONCLUSIONS: A protocol for nonmyeloablative allogeneic hematopoietic stem-cell transplantation that includes total-body irradiation and treatment with alemtuzumab and sirolimus can achieve stable, mixed donor-recipient chimerism and reverse the sickle cell phenotype. (ClinicalTrials.gov number, NCT00061568.) PMID- 20007562 TI - Images in clinical medicine. Blurred vision after cardiac catheterization. PMID- 20007561 TI - Telomere diseases. PMID- 20007563 TI - Case Records of the Massachusetts General Hospital. Case 38-2009 - a 16-year-old boy with paroxysmal headaches and visual changes. PMID- 20007564 TI - Hematopoietic stem-cell transplantation for adults with sickle cell disease. PMID- 20007565 TI - Ticagrelor versus clopidogrel in acute coronary syndromes. PMID- 20007566 TI - Intestinal transplantation. PMID- 20007567 TI - The Prometheus Payment model. PMID- 20007568 TI - Long-term follow-up of survival in Hodgkin's lymphoma. PMID- 20007569 TI - Comment on "Gene disruption study reveals a nonredundant role for TRIM21/Ro52 in NF-kappa B-dependent cytokine expression in fibroblasts". PMID- 20007570 TI - Inflammasome-associated nucleotide-binding domain, leucine-rich repeat proteins and inflammatory diseases. AB - The nucleotide-binding domain, leucine-rich repeat (NLR) proteins are a recently discovered family of intracellular pathogen and danger signal sensors. NLRs have emerged as important contributors to innate immunity in animals. The physiological impact of these genes is increasingly evident, underscored by the genetic association of variant family members with an array of inflammatory diseases. The association of mutations in NLR genes with autoinflammatory diseases indicates an important function of these genes in inflammation in vivo. This review summarizes the role of the inflammasome NLR proteins in innate immunity and inflammatory diseases and explores the possible utility of some of these NLRs as pharmacological targets. PMID- 20007571 TI - Asynchronous RAG-1 expression during B lymphopoiesis. AB - Changes in cell surface markers and patterns of gene expression are commonly used to construct sequences of events in hematopoiesis. However, the order may not be as rigid as once thought and it is unclear which changes represent the best milestones of differentiation. We developed a fate-mapping model where cells with a history of RAG-1 expression are permanently marked by red fluorescence. This approach is valuable for appreciating lymphoid-lineage relationships without need for irradiation and transplantation. Hematopoietic stem cells (HSC) as well as myeloid and dendritic cell progenitors were unlabeled. Also as expected, most previously identified RAG-1(+) early lymphoid progenitors in bone marrow and all lymphoid-affiliated cells were marked. Of particular interest, there was heterogeneity among canonical common lymphoid progenitors (CLP) in bone marrow. Labeled CLP expressed slightly higher levels of IL-7Ralpha, displayed somewhat less c-Kit, and generated CD19(+) lymphocytes faster than the unlabeled CLP. Furthermore, CLP with a history of RAG-1 expression were much less likely to generate dendritic and NK cells. The RAG-1-marked CLP were lineage stable even when exposed to LPS, while unlabeled CLP were redirected to become dendritic cells in response to this TLR4 ligand. These findings indicate that essential events in B lymphopoiesis are not tightly synchronized. Some progenitors with increased probability of becoming lymphocytes express RAG-1 while still part of the lineage marker-negative Sca-1(+)c-Kit(high) (LSK) fraction. Other progenitors first activate this locus after c-Kit levels have diminished and cell surface IL 7 receptors are detectable. PMID- 20007572 TI - IL-13R alpha 2 membrane and soluble isoforms differ in humans and mice. AB - Although mice have nanogram per milliliter serum levels of soluble (s) IL 13Ralpha2, humans lack sIL-13Ralpha2 in serum. Our data provide a mechanism for this biological divergence. In mice, discrete transcripts encoding soluble and membrane forms of IL-13Ralpha2 are generated by alternative splicing. We used small interfering RNA to specifically deplete the transcript encoding membrane (mem) IL-13Ralpha2 (full-length) or sIL-13Ralpha2 (DeltaEx10) in murine cells. Depletion of the full-length transcript decreased memIL-13Ralpha2 but had no effect on the level of sIL-13Ralpha2 in cell supernatants at baseline or following cytokine stimulation. Depletion of the DeltaEx10 transcript decreased sIL-13Ralpha2 in supernatants at baseline and following stimulation. In contrast to mice, we were unable to find a transcript encoding sIL-13Ralpha2 in humans and siRNA-mediated depletion of full-length IL-13Ralpha2 decreased both sIL-13Ralpha2 and memIL-13Ralpha2 in human cells. Inhibition of matrix metalloproteinases (MMP)/MMP-8 abolished production of sIL-13Ralpha2 from human cells. Thus, sIL 13Ralpha2 is derived exclusively from the memIL-13Ralpha2 transcript in humans through MMPs/MMP-8 cleavage of memIL-13Ralpha2, supporting a limited role for sIL 13Ralpha2 in humans and highlighting the potential importance of memIL-13Ralpha2 in human immunity. These observations require consideration when results of murine IL-13 studies are applied to humans. PMID- 20007574 TI - Dual control of antitumor CD8 T cells through the programmed death-1/programmed death-ligand 1 pathway and immunosuppressive CD4 T cells: regulation and counterregulation. AB - Tumors have evolved multiple mechanisms to evade immune destruction. One of these is expression of T cell inhibitory ligands such as programmed death-ligand 1 (PD L1; B7-H1). In this study, we show that PD-L1 is highly expressed on mesothelioma tumor cells and within the tumor stroma. However, PD-L1 blockade only marginally affected tumor growth and was associated with the emergence of activated programmed death-1(+) ICOS(+) CD4 T cells in tumor-draining lymph nodes, whereas few activated CD8 T cells were present. Full activation of antitumor CD8 T cells, characterized as programmed death-1(+) ICOS(+) Ki-67(+) and displaying CTL activity, was only observed when CD4 T cells were depleted, suggesting that a population of suppressive CD4 T cells exists. ICOS(+) foxp3(+) regulatory T cells were found to be regulated through PD-L1, identifying one potentially suppressive CD4 T cell population. Thus, PD-L1 blockade activates antitumor CD8 T cell most potently in the absence of CD4 T cells. These findings have implications for the development of PD-L1-based therapies. PMID- 20007573 TI - Neuronal I kappa B kinase beta protects mice from autoimmune encephalomyelitis by mediating neuroprotective and immunosuppressive effects in the central nervous system. AB - Some aspects of CNS-directed autoimmunity in multiple sclerosis are modeled in mice by immunization with myelin Ags where tissue damage is driven by myelin reactive Th1 and Th17 effector lymphocytes. Whether the CNS plays an active role in controlling such autoimmune diseases is unknown. We used mice in which IkappaB kinase beta was deleted from Ca(2+)/calmodulin-dependent kinase IIalpha expressing neurons (nIKKbetaKO) to investigate the contribution of neuronal NF kappaB to the development of myelin oligodendrocyte glycoprotein 35-55-induced experimental autoimmune encephalomyelitis. We show that nIKKbetaKO mice developed a severe, nonresolving disease with increased axon loss compared with controls and this was associated with significantly reduced CNS production of neuroprotective factors (vascular endothelial growth factor, CSF1-R, and FLIP) and increased production of proinflammatory cytokines (IL-6, TNF, IL-12, IL-17, and CD30L) and chemokines. The isolation of CNS-infiltrating monocytes revealed greater numbers of CD4(+) T cells, reduced numbers of NK1.1(+) cells, and a selective accumulation of Th1 cells in nIKKbetaKO CNS from early in the disease. Our results show that neurons play an important role in determining the quality and outcome of CNS immune responses, specifically that neuronal IkappaB kinase beta is required for neuroprotection, suppression of inflammation, limitation of Th1 lymphocyte accumulation, and enhancement of NK cell recruitment in experimental autoimmune encephalomyelitis-affected CNS and stress the importance of neuroprotective strategies for the treatment of multiple sclerosis. PMID- 20007575 TI - Involvement of the NLRP3 inflammasome in innate and humoral adaptive immune responses to fungal beta-glucan. AB - Fungal beta-glucan, such as curdlan, triggers antifungal innate immune responses as well as shaping adaptive immune responses. In this study, we identified a key pathway that couples curdlan to immune responses. Curdlan promoted the production of the proinflammatory cytokine IL-1beta by dendritic cells and macrophages through the NLRP3 inflammasome. Stimulation with Candida albicans and Saccharomyces cerevisiae also triggered the NLRP3 inflammasome-mediated IL-1beta production. In vivo, NLRP3 was required for efficient Ag-specific Ab production when curdlan was used as an adjuvant, whereas it was dispensable for the induction of Th1 and Th17 cell differentiation. Furthermore, stimulation of purified B cells with curdlan-induced CD69 up-regulation and IgM production while stimulation with other NLRP3 inflammasome activators, such as silica and aluminum salt, did not. Notably, this induction required NLRP3 but was independent of Toll like receptor and IL-1 receptor family signaling, suggesting the presence of NLRP3-dependent and IL-1 receptor family independent mechanisms in B cells responsible for Ab responses. Collectively, these findings reveal a critical role for the NLRP3 inflammasome in the regulation of antifungal innate immune responses as well as B cell activation. PMID- 20007577 TI - Helicobacter pylori induces MAPK phosphorylation and AP-1 activation via a NOD1 dependent mechanism. AB - Helicobacter pylori rapidly activates MAPKs and transcription factors, NF-kappaB and AP-1, in gastric epithelial cells following host attachment. Activation of these signal transducers is largely dependent on the cag pathogenicity island (cagPAI)-encoded Type IV Secretion System. H. pylori was shown to translocate peptidoglycan through the Type IV Secretion System, which is recognized by the pathogen recognition molecule, NOD1, thus resulting in NF-kappaB activation. The mechanisms of H. pylori-induced MAPK and AP-1 activation, however, are less well defined and therefore, we assessed the contribution of NOD1 to their activation. For this, we used gastric epithelial cell lines, stably expressing siRNA to either NOD1 or a control gene. In siNOD1-expressing cells stimulated with cagPAI(+) H. pylori, we observed significant reductions in p38 and ERK phosphorylation (p < 0.05), whereas the levels of Jnk phosphorylation remained unchanged. Consistent with a previous report, however, we were able to demonstrate NOD1-dependent Jnk phosphorylation by the invasive pathogen Shigella flexneri, highlighting pathogen-specific host responses to infection. We also show that NOD1 was essential for H. pylori induction of not only NF-kappaB, but also AP-1 activation, implying that NOD1 induces robust proinflammatory responses, in an attempt to rapidly control infection. Pharmacological inhibition of p38 and ERK activity significantly reduced IL-8 production in response to H. pylori, further emphasizing the importance of MAPKs in innate immune responses to the pathogen. Thus, for the first time we have shown the important role for NOD1 in MAPK and AP-1 activation in response to cagPAI(+) H. pylori. PMID- 20007576 TI - Inflation and long-term maintenance of CD8 T cells responding to a latent herpesvirus depend upon establishment of latency and presence of viral antigens. AB - Following the priming and contraction phases of the T cell response, latent persistent herpesviruses lead to an accumulation of large pools of virus-specific CD8 T cells, also known as memory inflation (MI). The mechanism of this inflation is incompletely understood, largely because the molecular reactivation of these viruses in vivo and its impact upon T cell biology have not been resolved in mice, and because the relevant observations in humans remain, by necessity, correlative. Understanding these processes is essential from the standpoint of the proposed critical role for latent herpesviruses in aging of the immune system. We studied the causes of memory CD8 T cell accumulation following systemic HSV-1 administration as a model of widespread latent viral infection in humans. A direct role of viral latency and Ag-specific restimulation in driving the accumulation and maintenance of inflated CD8 T cells and a strongly suggested role of viral reactivation in that process were shown by the following: 1) lack of MI in the absence of established latency; 2) prevention or delay of MI with drugs that curtail viral replication; and 3) abrogation of MI by the transfer of inflated T cells into a virus-free environment. These results strongly suggest that periodic, subclinical reactivations of a latent persistent virus cause dysregulation of memory CD8 T cell homeostasis, similar to the one in humans. Moreover, results with antiviral drugs suggest that this approach could be considered as a treatment modality for maintaining T cell diversity and/or function in old age. PMID- 20007579 TI - Cell surface externalization of annexin A1 as a failsafe mechanism preventing inflammatory responses during secondary necrosis. AB - The engulfment of apoptotic cells is of crucial importance for tissue homeostasis in multicellular organisms. A failure of this process results in secondary necrosis triggering proinflammatory cytokine production and autoimmune disease. In the present study, we investigated the role of annexin A1, an intracellular protein that has been implicated in the efficient removal of apoptotic cells. Consistent with its function as bridging protein in the phagocyte synapse, opsonization of apoptotic cells with purified annexin A1 strongly enhanced their phagocytic uptake. A detailed analysis, however, surprisingly revealed that annexin A1 was hardly exposed to the cell surface of primary apoptotic cells, but was strongly externalized only on secondary necrotic cells. Interestingly, while the exposure of annexin A1 failed to promote the uptake of these late secondary necrotic cells, it efficiently prevented induction of cytokine production in macrophages during engulfment of secondary necrotic cells. Our results therefore suggest that annexin A1 exposure during secondary necrosis provides an important failsafe mechanism counteracting inflammatory responses, even when the timely clearance of apoptotic cells has failed. PMID- 20007578 TI - Novel role for aldose reductase in mediating acute inflammatory responses in the lung. AB - Exaggerated inflammatory responses and the resultant increases in alveolar capillary permeability underlie the pathogenesis of acute lung injury during sepsis. This study examined the functions of aldose reductase (AR) in mediating acute lung inflammation. Transgenic mice expressing human AR (ARTg) were used to study the functions of AR since mice have low intrinsic AR activity. In a mild cecal ligation and puncture model, ARTg mice demonstrated an enhanced AR activity and a greater inflammatory response as evaluated by circulating cytokine levels, neutrophil accumulation in the lungs, and activation of Rho kinase in lung endothelial cells (ECs). Compared with WT lung cells, ARTg lung cells produced more IL-6 and showed augmented JNK activation in response to LPS stimulation ex vivo. In human neutrophils, AR activity was required for fMLP-included CD11b activation and up-regulation, respiratory burst, and shape changes. In human pulmonary microvascular ECs, AR activity was required for TNF-alpha-induced activation of the Rho kinase/MKK4/JNK pathway and IL-6 production, but not p38 activation or ICAM-1 expression. Importantly, AR activity in both human neutrophils and ECs was required for neutrophil adhesion to TNF-alpha-stimulated ECs. These data demonstrate a novel role for AR in regulating the signaling pathways leading to neutrophil-EC adhesion during acute lung inflammation. PMID- 20007580 TI - Decreased levels of secretory leucoprotease inhibitor in the Pseudomonas-infected cystic fibrosis lung are due to neutrophil elastase degradation. AB - Secretory leucoprotease inhibitor (SLPI) is a neutrophil serine protease inhibitor constitutively expressed at many mucosal surfaces, including that of the lung. Originally identified as a serine protease inhibitor, it is now evident that SLPI also has antimicrobial and anti-inflammatory functions, and therefore plays an important role in host defense. Previous work has shown that some host defense proteins such as SLPI and elafin are susceptible to proteolytic degradation. Consequently, we investigated the status of SLPI in the cystic fibrosis (CF) lung. A major factor that contributes to the high mortality rate among CF patients is Pseudomonas aeruginosa infection. In this study, we report that P. aeruginosa-positive CF bronchoalveolar lavage fluid, which contains lower SLPI levels and higher neutrophil elastase (NE) activity compared with P. aeruginosa-negative samples, was particularly effective at cleaving recombinant human SLPI. Additionally, we found that only NE inhibitors were able to prevent SLPI cleavage, thereby implicating NE in this process. NE in excess was found to cleave recombinant SLPI at two novel sites in the NH(2)-terminal region and abrogate its ability to bind LPS and NF-kappaB consensus binding sites but not its ability to inhibit activity of the serine protease cathepsin G. In conclusion, this study provides evidence that SLPI is cleaved and inactivated by NE present in P. aeruginosa-positive CF lung secretions and that P. aeruginosa infection contributes to inactivation of the host defense screen in the CF lung. PMID- 20007581 TI - DcR3 protects islet beta cells from apoptosis through modulating Adcyap1 and Bank1 expression. AB - The islet primary nonfunction (PNF) is a serious problem in islet transplantation. In this study, we investigated whether DcR3-secreting transgenic (Tg) islets could reduce PNF. We generated Tg mice expressing human DcR3. The transgenically expressed DcR3 protected islets from IFN-gamma plus IL-1beta- or TNF-alpha plus IL-1beta-induced dysfunction and apoptosis in vitro. The Tg islets presented significantly reduced PNF after transplantation. Mechanistically, in addition to the known FasL apoptotic pathway, components of two other apoptosis pathways, that is, HVEM/LTbetaR for the LIGHT pathway and DR3 for the TL1A pathway, were found to be expressed in islets. Recombinant LIGHT- and TL1A induced islet apoptosis in the absence of the FasL/Fas pathway, as well as DcR3, could block such induction. These results for the first time demonstrated that LIGHT and TL1A were capable of inducing islet apoptosis in addition to FasL, while DcR3 protected the islets by blocking all three apoptosis pathways. By DNA microarray analysis, we discovered that Adcyap was up-regulated >700-fold and Bank1 was down-regulated 50-fold in the cytokine-assaulted Tg islets, compared with WT islets. Forced overexpression of Adcyap1 by plasmid transfection or knockdown of Bank1 expression by small interfering RNA in insulinoma NIT-1 cells protected them from cytokine-triggered apoptosis, indicating that indeed DcR3 protects beta cells via the action of these two downstream molecules. This study has revealed novel mechanisms by which DcR3 protects islet survival, and it has identified new therapeutic targets of diabetes. PMID- 20007582 TI - Inhibition of IL-17A attenuates atherosclerotic lesion development in apoE deficient mice. AB - The importance of an (auto)immune response in atherogenesis is becoming increasingly well understood. IL-17A-expressing T cells modulate immune cell trafficking, initiating inflammation and cytokine production in (auto)immune diseases. In human carotid artery plaques, we previously showed the presence of IL-17A-producing T cells and IL-23; however, IL-17A effects on atherogenesis have not been studied. Aortic root sections from 8-wk-old apolipoprotein E-deficient mice fed a standard chow diet were examined after 12 wk for lesion area, plaque composition, cellular infiltration, cytokine expression, and apoptosis. The treatment group (n = 15) received anti-IL-17A Ab and the controls (n = 10) received irrelevant Abs. Inhibition of IL-17A markedly reduced atherosclerotic lesion area (p < 0.001), maximal stenosis (p < 0.001), and vulnerability of the lesion. IL-17A mAb-treated mice showed reduced cellular infiltration, down regulation of activation markers on endothelium and immune cells (e.g., VCAM-1), and reduced cytokine/chemokine secretion (e.g., IL6, TNFalpha, CCL5). To investigate possible mechanisms, different atherogenic cell types (e.g., macrophages, dendritic cells, HUVECs, vascular smooth muscle cells) were stimulated with IL-17A in addition to TNF-alpha, IFN-gamma, or LPS to induce cellular activation or apoptosis in vitro. Stimulation with IL-17A induced proinflammatory changes in several atherogenic cell types and apoptotic cell death in murine cells. Functional blockade of IL-17A reduces atherosclerotic lesion development and decreases plaque vulnerability, cellular infiltration, and tissue activation in apolipoprotein E-deficient mice. The present data support a pathogenic role of IL-17A in the development of atherosclerosis by way of its widespread proinflammatory and proapoptotic effects on atherogenic cells. PMID- 20007583 TI - Expression of soluble vascular endothelial growth factor receptor-1 in human monocyte-derived mature dendritic cells contributes to their antiangiogenic property. AB - The soluble form of vascular endothelial growth factor receptor-1 (sVEGFR-1) is produced from endothelial cells by alternative splicing of VEGFR-1 mRNA, and can inhibit angiogenesis by blocking the biological effects of VEGF. In this study, we show the expression of a large amount of sVEGFR-1 in human monocyte-derived mature dendritic cells (mDCs). As compared with monocytes and immature DCs, mDCs generated by TNF-alpha or soluble CD40L with IFN-gamma, but not LPS or other stimuli, preferentially produce sVEGFR-1. We also detected the mRNA of sVEGFR-1 generated by alternative splicing of VEGFR-1 mRNA in mDCs induced by TNF-alpha. The production of sVEGFR-1 showed a distinct contrast to those of VEGF in each DC matured with various stimuli. The supernatant of DCs matured with TNF-alpha or soluble CD40L with IFN-gamma showed inhibition of the tube formation of HUVECs, which was neutralized by anti-VEGFR-1 Ab, indicating that sVEGFR-1 secreted from mDCs was biologically active. Interestingly, the supernatant of mDCs generated with LPS increased HUVEC capillary-like formation in vitro. The ratio of sVEGFR-1 to VEGF clearly reflected the net angiogenic property of mDCs. Administration of mDCs induced by TNF-alpha into the s.c. tumor of PC-14 cells implanted in SCID mice demonstrated the inhibition of tumor growth via reduction of the number of CD31-positive vessels, indicating their in vivo antiangiogenic potential. These results suggest that sVEGFR-1 produced by mDCs contribute to their antiangiogenic property, and the ratio of sVEGFR-1 to VEGF might be a useful tool for evaluating their ability to regulate angiogenesis mediated by VEGF. PMID- 20007584 TI - IL-1R1/MyD88 signaling is critical for elastase-induced lung inflammation and emphysema. AB - Lung emphysema and fibrosis are severe complications of chronic obstructive pulmonary disease, and uncontrolled protease activation may be involved in the pathogenesis. Using experimental elastase-induced acute inflammation, we demonstrate here that inflammation and development of emphysema is IL-1R1 and Toll/IL-1R signal transduction adaptor MyD88 dependent; however, TLR recognition is dispensable in this model. Elastase induces IL-1beta, TNF-alpha, keratinocyte derived chemokine, and IL-6 secretion and neutrophil recruitment in the lung, which is drastically reduced in the absence of IL-1R1 or MyD88. Further, tissue destruction with emphysema and fibrosis is attenuated in the lungs of IL-1R1- and MyD88-deficient mice. Specific blockade of IL-1 by IL-1R antagonist diminishes acute inflammation and emphysema. Finally, IL-1beta production and inflammation are reduced in mice deficient for the NALP3 inflammasome component apoptosis associated speck-like protein containing a caspase recruitment domain (ASC), and we identified uric acid, which is produced upon elastase-induced lung injury, as an activator of the NALP3/ASC inflammasome. In conclusion, elastase-mediated lung pathology depends on inflammasome activation with IL-1beta production. IL-1beta therefore represents a critical mediator and a possible therapeutic target of lung inflammation leading to emphysema. PMID- 20007586 TI - Autoantibodies against C1q in systemic lupus erythematosus are antigen-driven. AB - Autoantibodies against complement C1q (anti-C1q Abs) were shown to strongly correlate with the occurrence of severe nephritis in patients with systemic lupus erythematosus (SLE), suggesting a potential pathogenic role by interfering with the complement cascade. To analyze the humoral immune response against C1q at the molecular level, we screened a bone marrow-derived IgGkappa/IgGlambda Fab phage display library from a SLE patient with high anti-C1q Ab titer against purified human C1q. Six Fabs that exhibited strong binding to C1q in ELISA were isolated. The anti-C1q Fabs recognized neoepitopes that were only exposed on bound C1q and not present on soluble C1q mapping to different regions of the collagen-like region of C1q. Analysis of the genes encoding the variable H and L chains of the IgG-derived anti-C1q Fab revealed that all the variable H and L chain regions were highly mutated, with nucleotide and amino acid homologies to the closest germline in the range of 71-97% (average 85 +/- 4) and 72-92% (average 88 +/- 6), respectively. In addition, the variable region of the Fabs exhibited high replacement to silent ratios. The six anti-C1q Fabs were shown to be of high affinity, with a K(d) ranging from of 8.4 x 10(-8) M to 1.4 x 10(-7) M, comparable to an antiviral immune response. Our data underlines the notion that the development of anti-C1q Abs in SLE is the consequence of an Ag-driven, affinity-matured immune response. Those anti-C1q Fabs are unique tools to address how complement C1q is implicated in the pathogenesis of SLE. PMID- 20007585 TI - Dynamics of the interaction of human IgG subtype immune complexes with cells expressing R and H allelic forms of a low-affinity Fc gamma receptor CD32A. AB - CD32A, the major phagocytic FcgammaR in humans, exhibits a polymorphism in the ligand binding domain. Individuals homozygous for the R allelic form of CD32A (CD32A(R) allele) are more susceptible to bacterial infections and autoimmune diseases as compared with H allelic CD32A (CD32A(H)) homozygous and CD32A(R/H) heterozygous individuals. To understand the mechanisms behind this differential susceptibility, we have investigated the dynamics of the interaction of these allelic forms of CD32A when they are simultaneously exposed to immune complexes (IC). Binding studies using Ig fusion proteins of CD32A alleles showed that the R allele has significantly lower binding not only to human IgG2, but also to IgG1 and IgG3 subtypes. Competition assays using purified molecules demonstrated that CD32A(H)-Ig outcompetes CD32A(R)-Ig for IC binding when both alleles simultaneously compete for the same ligand. CD32A(H)-Ig blocked the IC binding mediated by both the allelic forms of cell surface CD32A, whereas CD32A(R)-Ig blocked only CD32A(R) and was unable to cross-block IC binding mediated by CD32A(H). Two-dimensional affinity measurements also demonstrated that CD32A(R) has significantly lower affinity toward all three subtypes as compared with CD32A(H). Our data suggest that the lower binding of CD32A(R) not only to IgG2 but also to IgG1 and IgG3 might be responsible for the lack of clearance of IC leading to increased susceptibility to bacterial infections and autoimmune diseases. Our data further suggests that in humans, inflammatory cells from CD32A(R/H) heterozygous individuals may predominantly use the H allele to mediate Ab-coated target cell binding during phagocytosis and Ab-dependent cellular cytotoxicity, resulting in a phenotype similar to CD32A(H) homozygous individuals. PMID- 20007587 TI - In vivo induction of type 1-like regulatory T cells using genetically modified B cells confers long-term IL-10-dependent antigen-specific unresponsiveness. AB - Regulatory T cells (Tregs) hold much promise for the therapy of allergy and autoimmunity, but their use is hampered by lack of Ag specificity (natural Tregs) and difficulty to expand in vitro or in vivo (adaptive Tregs). We designed a method for in vivo induction of Ag-specific Tregs, in BALB/c H-2d, that share characteristics with type 1 Tregs (Tr1). A retroviral vector was constructed encoding a major T cell epitope of a common allergen, Der p 2, fused to an endosomal targeting sequence (gp75) for efficient MHC class II presentation. B cells transduced with such construct were adoptively transferred to BALB/c mice before or after peptide immunization. Long-lasting Ag-specific immune tolerance was achieved in both cases. Genetically modified B cells constitutively expressed the transgene for at least 3 mo. B cells from IL-10(-/-) mice were unable to induce tolerance. Upon transfer, B cells induced Foxp3(-)CD4(+) T cells showing phenotypic and functional characteristics comparable to Tr1-cells, including production of IL-10 but not of TGF-beta, and high expression of CTLA-4. Adoptive transfer of such T cells conferred unresponsiveness to allergen immunization and prevented the development of Der p 2-induced asthma. Functional Tr1-like cells can therefore be induced in vivo using retrovirally transduced B cells. PMID- 20007588 TI - Differential role of the Fas/Fas ligand apoptotic pathway in inflammation and lung fibrosis associated with reovirus 1/L-induced bronchiolitis obliterans organizing pneumonia and acute respiratory distress syndrome. AB - Bronchiolitis obliterans organizing pneumonia (BOOP) and acute respiratory distress syndrome (ARDS) are two clinically and histologically distinct syndromes sharing the presence of an inflammatory and fibrotic component. Apoptosis via the Fas/Fas ligand (FasL) pathway plays an important role in the development of acute lung injury and fibrosis characteristic of these and other pulmonary inflammatory and fibrotic syndromes. We evaluated the role of apoptosis via the Fas/FasL pathway in the development of pulmonary inflammation and fibrosis in reovirus 1/L induced BOOP and ARDS. CBA/J mice were intranasally inoculated with saline, 1 x 10(6) (BOOP), or 1 x 10(7) (ARDS) PFU reovirus 1/L, and evaluated at various days postinoculation for in situ apoptosis by TUNEL analysis and Fas/FasL expression. Our results demonstrate the presence of apoptotic cells and up-regulation of Fas/FasL expression in alveolar epithelium and in infiltrating cells during the inflammatory and fibrotic stages of both reovirus 1/L-induced ARDS and BOOP. Treatment of mice with the caspase 8 inhibitor, zIETD-fmk, inhibited apoptosis, inflammation, and fibrotic lesion development in reovirus 1/L-induced BOOP and ARDS. However, CBA/KlJms-Fas(lpr-cg)/J mice, which carry a point mutation in the Fas cytoplasmic region that abolishes the ability of Fas to transduce an apoptotic signal, do not develop pulmonary inflammation and fibrotic lesions associated with reovirus 1/L-induced BOOP, but still develop inflammation and fibrotic lesions associated with reovirus 1/L-induced ARDS. These results suggest a differential role for the Fas/FasL apoptotic pathway in the development of inflammation and fibrotic lesions associated with BOOP and ARDS. PMID- 20007589 TI - ERK-dependent T cell receptor threshold calibration in rheumatoid arthritis. AB - Immune responses to citrullinated neoantigens and clinical efficacy of costimulation blockade indicate a general defect in maintaining T cell tolerance in rheumatoid arthritis (RA). To examine whether TCR threshold calibration contributes to disease pathogenesis, signaling in RA T cells was quantified. RA patients had a selective increase in ERK phosphorylation compared with demographically matched controls due to a mechanism distal of Ras activation. Increased ERK responses included naive and memory CD4 and CD8 T cells and did not correlate with disease activity. The augmented ERK activity delayed SHP-1 recruitment to the TCR synapse and sustained TCR-induced Zap70 and NF-kappaB signaling, facilitating responses to suboptimal stimulation. Increased responsiveness of the ERK pathway was also a characteristic finding in the SKG mouse model of RA where it preceded clinical symptoms. Treatment with subtherapeutic doses of a MEK-1/2 inhibitor delayed arthritis onset and reduced severity, suggesting that increased ERK phosphorylation predisposes for autoimmunity and can be targeted to prevent disease. PMID- 20007590 TI - A dual action of rheumatoid arthritis synovial fibroblast IL-15 expression on the equilibrium between CD4+CD25+ regulatory T cells and CD4+CD25- responder T cells. AB - We previously described that fibroblast-like cells from the synovium of rheumatoid arthritis patients (RASFib) constitutively express intracellular and surface IL-15, which induces activation of cocultured T cells. Our objective was to study the effect of RASFib IL-15 expression on the function of human CD4(+)CD25(+) regulatory T cells (Treg). RASFib, through their constitutive IL-15 expression, were able to induce the proliferation of human Tregs stimulated through their TCR, and at the same time potentiated their suppressive action on the cytokine secretion of CD4(+)CD25(-) responder T cells (Tresp). In parallel, constitutive RASFib IL-15 expression mediated an up-regulated response of Tresp. Subsequently, total CD4(+) T cells, containing natural proportions of Treg and Tresp, secreted an increased amount of pathogenic cytokines when cocultured with RASFib despite the presence of proliferating Treg with superior regulatory potency. In summary, RASFib IL-15 exerts a dual action on the equilibrium between Treg and Tresp by potentiating the suppressive effect of Treg while augmenting the proinflammatory action of Tresp; the result is a shift of the Treg/Tresp balance toward a proinflammatory state. This alteration of the Treg/Tresp equilibrium is not observed in the presence of osteoarthritis synovial fibroblasts or dermal fibroblasts, which do not constitutively express surface IL 15. Additionally, Treg with superior suppressive potency were present in the peripheral blood and the synovial fluid of RA patients, but this enhanced immunoregulatory activity was not able to overcome the increased secretion of pathogenic cytokines by RA-Tresp, indicating that rheumatoid arthritis patients demonstrate an altered Treg/Tresp equilibrium in vivo. PMID- 20007591 TI - Allele *1 of HS1.2 enhancer associates with selective IgA deficiency and IgM concentration. AB - Selective IgA deficiency (IGAD) is the most common primary immunodeficiency, yet its pathogenesis is elusive. The IG (heavy) H chain human 3' Regulatory Region harbors three enhancers and has an important role in Ig synthesis. HS1.2 is the only polymorphic enhancer of the 3' RRs. We therefore evaluated HS1.2 allelic frequencies in 88 IGAD patients and 101 controls. Our data show that IGAD patients have a highly significant increase of homozygousity of the allele *1 (39% in the IGAD patients and 15% in controls), with an increase of 2.6-fold. Allele *4 has a similar trend of allele *2, both showing a significant decrease of frequency in IGAD. No relationship was observed between allele *1 frequencies and serum levels of IgG. However, allele *1 was associated in IGAD patients with relatively low IgM levels (within the 30th lowest percentile of patients). The HS1.2 polymorphism influences Ig seric production, but not IgG switch, in fact 30th lowest or highest percentile of IgG in patients did not associate to different frequencies of HS1.2 alleles. The control on normal healthy subjects did not correlate high or low levels of IgM or IgG with HS1.2 allelic frequence variation. Overall our candidate gene approach confirms that the study of polymorphisms in human diseases is a valid tool to investigate the function of these Regulatory Regions that confers multiple immune features. PMID- 20007592 TI - The kinase inhibitors sunitinib and sorafenib differentially affect NK cell antitumor reactivity in vitro. AB - Sunitinib and Sorafenib are protein kinase inhibitors (PKI) approved for treatment of patients with advanced renal cell cancer (RCC). However, long-term remissions of advanced RCC have only been observed after IL-2 treatment, which underlines the importance of antitumor immune responses in RCC patients. Because PKI, besides affecting tumor cells, also may inhibit signaling in immune effector cells, we determined how Sunitinib and Sorafenib influence antitumor immunity. We found that cytotoxicity and cytokine production of resting and IL-2-activated PBMC are inhibited by pharmacological concentrations of Sorafenib but not Sunitinib. Analysis of granule-mobilization within PBMC revealed that this was due to impaired reactivity of NK cells, which substantially contribute to antitumor immunity by directly killing target cells and shaping adaptive immune responses by secreting cytokines like IFN-gamma. Analyses with resting and IL-2 activated NK cells revealed that both PKI concentration dependently inhibit cytotoxicity and IFN-gamma production of NK cells in response to tumor targets. This was due to impaired PI3K and ERK phosphorylation which directly controls NK cell reactivity. However, while Sorafenib inhibited NK cell effector functions and signaling at levels achieved upon recommended dosing, pharmacological concentrations of Sunitinib had no effect, and this was observed upon stimulation of NK cell reactivity by tumor target cells and upon IL-2 treatment. In light of the important role of NK cells in antitumor immunity, and because multiple approaches presently aim to combine PKI treatment with immunotherapeutic strategies, our data demonstrate that choice and dosing of the most suitable PKI in cancer treatment requires careful consideration. PMID- 20007595 TI - Cell to cell transfer of the chromatin-packaged human beta-globin gene cluster. AB - Cell type-specific gene expression is regulated by chromatin structure and the transcription factors provided by the cells. In the present study, we introduced genes packaged into chromatin into target cells using a human artificial chromosome (HAC) and analyzed regulation of gene expression. The human beta globin gene cluster was built into an HAC (globin-HAC) and introduced into mouse embryonic stem (ES) cells using microcell-mediated chromosome transfer (MMCT); the adult-type human beta-globin gene was expressed in bone marrow and spleen cells of the transgenic mice. In vitro differentiation of ES cells into mouse erythrocytes indicated that the natural sequential expression of epsilon, gamma and beta-globin genes was reproduced on the globin-HAC. Combination of MMCT and a novel chromosome transfection technique allowed transfer of globin-HAC from HT1080 cells into the human leukemia cell line K562, and from K562 cells back into HT1080 cells. Expression of the gamma-globin gene, repressed in HT1080 cells, was activated in K562 cells without any processes of differentiation into adult erythroid cells, and was completely repressed again in HT1080 cells when transferred back from K562 cells. Thus, transfer of target genes packaged into chromatin using a HAC was useful for functional analyses of gene regulation. PMID- 20007597 TI - A cassette of N-terminal amino acids of histone H2B are required for efficient cell survival, DNA repair and Swi/Snf binding in UV irradiated yeast. AB - The highly charged histone N-terminal domains are engaged in inter- and intra nucleosomal interactions, and contain a host of sites used for posttranslational modification. We have studied the effect of deleting residues 30-37 from the N terminal domain of histone H2B in yeast cells, on nucleotide excision repair (NER) following UV irradiation, as these cells are quite sensitive to UV. We find that H2B Delta30-37 cells exhibit reduced NER efficiency at three specific chromatin loci: the transcriptionally active, RPB2 locus; the transcriptionally silenced, nucleosome-loaded HML locus; and the transcriptionally repressed, non silenced, GAL10 locus. Nuclease digestion studies indicate that H2B Delta30-37 chromatin has increased nucleosome accessibility and/or nucleosome mobility. In addition, H2B Delta30-37 mutants acquire more DNA damage, compared to wt cells, following the same dose of UV radiation. Reducing the level of damage in H2B Delta30-37 cells to match that of wt cells restores the NER rate to wt levels in the RPB2 and GAL10 loci, but NER efficiency remains low in the silenced HML locus. Interestingly, recruitment of Snf5 to the HML locus is reduced in H2B Delta30-37 cells and more transient following UV irradiation. This may reflect a lower binding affinity of the SWI/SNF complex to H2B Delta30-37 nucleosomes. PMID- 20007596 TI - The structure of the KlcA and ArdB proteins reveals a novel fold and antirestriction activity against Type I DNA restriction systems in vivo but not in vitro. AB - Plasmids, conjugative transposons and phage frequently encode anti-restriction proteins to enhance their chances of entering a new bacterial host that is highly likely to contain a Type I DNA restriction and modification (RM) system. The RM system usually destroys the invading DNA. Some of the anti-restriction proteins are DNA mimics and bind to the RM enzyme to prevent it binding to DNA. In this article, we characterize ArdB anti-restriction proteins and their close homologues, the KlcA proteins from a range of mobile genetic elements; including an ArdB encoded on a pathogenicity island from uropathogenic Escherichia coli and a KlcA from an IncP-1b plasmid, pBP136 isolated from Bordetella pertussis. We show that all the ArdB and KlcA act as anti-restriction proteins and inhibit the four main families of Type I RM systems in vivo, but fail to block the restriction endonuclease activity of the archetypal Type I RM enzyme, EcoKI, in vitro indicating that the action of ArdB is indirect and very different from that of the DNA mimics. We also present the structure determined by NMR spectroscopy of the pBP136 KlcA protein. The structure shows a novel protein fold and it is clearly not a DNA structural mimic. PMID- 20007598 TI - RNA helicase A modulates translation of HIV-1 and infectivity of progeny virions. AB - Retroviruses rely on host RNA-binding proteins to modulate various steps in their replication. Previously several animal retroviruses were determined to mediate Dhx9/RNA helicase A (RHA) interaction with a 5' terminal post-transcriptional control element (PCE) for efficient translation. Herein PCE reporter assays determined HTLV-1 and HIV-1 RU5 confer orientation-dependent PCE activity. The effect of Dhx9/RHA down-regulation and rescue with siRNA-resistant RHA on expression of HIV-1(NL4-3) provirus determined that RHA is necessary for efficient HIV-1 RNA translation and requires ATPase-dependent helicase function. Quantitative analysis determined HIV-1 RNA steady-state and cytoplasmic accumulation were not reduced; rather the translational activity of viral RNA was reduced. Western blotting determined that RHA-deficient virions assemble with Lys tRNA synthetase, exhibit processed reverse transcriptase and contain similar level of viral RNA, but they are poorly infectious on primary lymphocytes and HeLa cells. The results demonstrate RHA is an important host factor within the virus-producer cell and within the viral particle. The identification of RHA dependent PCE activity in cellular junD RNA and in six of seven genera of Retroviridae suggests conservation of this translational control mechanism among vertebrates, and convergent evolution of Retroviridae to utilize this host mechanism. PMID- 20007599 TI - Frame-disrupting mutations elicit pre-mRNA accumulation independently of frame disruption. AB - The T-cell receptor (TCR) and immunoglobulin (Ig) genes are unique among vertebrate genes in that they undergo programmed rearrangement, a process that allows them to generate an enormous array of receptors with different antigen specificities. While crucial for immune function, this rearrangement mechanism is highly error prone, often generating frameshift or nonsense mutations that render the rearranged TCR and Ig genes defective. Such frame-disrupting mutations have been reported to increase the level of TCRbeta and Igmicro pre-mRNA, suggesting the hypothesis that RNA processing is blocked when frame disruption is sensed. Using a chimeric gene that contains TCRbeta sequences conferring this upregulatory response, we provide evidence that pre-mRNA upregulation is neither frame- nor translation-dependent; instead, several lines of evidence suggested that it is the result of disrupted cis elements necessary for efficient RNA splicing. In particular, we identify the rearranging VDJ(beta) exon as being uniquely densely packed with exonic-splicing enhancers (ESEs), rendering this exon hypersensitive to mutational disruption. As the chimeric gene that we developed for these studies generates unusually stable nuclear pre-mRNAs that accumulate when challenged with ESE mutations, we suggest it can be used as a sensitive in vivo system to identify and characterize ESEs. PMID- 20007600 TI - The N-terminus of Prp1 (Prp6/U5-102 K) is essential for spliceosome activation in vivo. AB - The spliceosomal protein Prp1 (Prp6/U5-102 K) is necessary for the integrity of pre-catalytic spliceosomal complexes. We have identified a novel regulatory function for Prp1. Expression of mutations in the N-terminus of Prp1 leads to the accumulation of pre-catalytic spliceosomal complexes containing the five snRNAs U1, U2, U5 and U4/U6 and pre-mRNAs. The mutations in the N-terminus, which prevent splicing to occur, include in vitro and in vivo identified phosphorylation sites of Prp4 kinase. These sites are highly conserved in the human ortholog U5-102 K. The results presented here demonstrate that structural integrity of the N-terminus is required to mediate a splicing event, but is not necessary for the assembly of spliceosomes. PMID- 20007601 TI - Heterodimeric DNA methyltransferases as a platform for creating designer zinc finger methyltransferases for targeted DNA methylation in cells. AB - The ability to target methylation to specific genomic sites would further the study of DNA methylation's biological role and potentially offer a tool for silencing gene expression and for treating diseases involving abnormal hypomethylation. The end-to-end fusion of DNA methyltransferases to zinc fingers has been shown to bias methylation to desired regions. However, the strategy is inherently limited because the methyltransferase domain remains active regardless of whether the zinc finger domain is bound at its cognate site and can methylate non-target sites. We demonstrate an alternative strategy in which fragments of a DNA methyltransferase, compromised in their ability to methylate DNA, are fused to two zinc fingers designed to bind 9 bp sites flanking a methylation target site. Using the naturally heterodimeric DNA methyltransferase M.EcoHK31I, which methylates the inner cytosine of 5'-YGGCCR-3', we demonstrate that this strategy can yield a methyltransferase capable of significant levels of methylation at the target site with undetectable levels of methylation at non-target sites in Escherichia coli. However, some non-target methylation could be detected at higher expression levels of the zinc finger methyltransferase indicating that further improvements will be necessary to attain the desired exclusive target specificity. PMID- 20007602 TI - Gene silencing by cell-penetrating, sequence-selective and nucleic-acid hydrolyzing antibodies. AB - Targeting particular mRNAs for degradation is a fascinating approach to achieve gene silencing. Here we describe a new gene silencing tool exploiting a cell penetrating, nucleic-acid hydrolyzing, single-domain antibody of the light-chain variable domain, 3D8 VL. We generated a synthetic library of 3D8 VL on the yeast surface by randomizing residues located in one of two beta-sheets. Using 18-bp single-stranded nucleic acids as target substrates, including the human Her2/neu targeting sequence, we selected 3D8 VL variants that had approximately 100-1000 fold higher affinity and approximately 2-5-fold greater selective hydrolyzing activity for target substrates than for off targets. 3D8 VL variants efficiently penetrated into living cells to be accumulated in the cytosol and selectively decreased the amount of target sequence-carrying mRNAs as well as the proteins encoded by these mRNAs with minimal effects on off-target genes. In particular, one 3D8 VL variant targeting the Her2 sequence showed more efficient downregulation of Her2 expression than a small-interfering RNA targeting the same Her2 sequence, resulting in apoptotic cell death of Her2-overexpressing breast cancer cells. Our results demonstrate that cell-penetrating 3D8 VL variants with sequence-selective, nucleic-acid-hydrolyzing activity can selectively degrade target mRNAs in the cytosol, providing a new gene silencing tool mediated by antibody. PMID- 20007603 TI - The rate of mutation of a single gene. AB - The rate of mutation refers to the probability that a unit length of DNA (generally a base pair) mutates with time. Fluctuation analysis or mutant accumulation assays applied to phenotypic changes measure mutation rates of cells. However, only a few phenotypic changes indicative of mutations are known thus limiting the analysis to those rare genes. Direct sequencing overcomes the limitations imposed by phenotypic analysis but is limited by the extensive number of clones or cells that have to be analyzed in fluctuation or mutant accumulation assays. We propose a strategy to determine the rate of mutation of a gene by limited direct sequencing of a few single cells of a defined lineage. To accomplish this, we determined the average number of mutations per position in each DNA length sequenced from the proportion of the non-mutated positions, according to the Poisson process and/or the Taylor series. Measuring the rate of mutation by direct sequencing of genes does not require ascertaining a phenotype and can be applied to any area of the genome in a cell. The approach avoids fluctuation errors. PMID- 20007604 TI - Rad26p, a transcription-coupled repair factor, is recruited to the site of DNA lesion in an elongating RNA polymerase II-dependent manner in vivo. AB - Rad26p, a yeast homologue of human Cockayne syndrome B with an ATPase activity, plays a pivotal role in stimulating DNA repair at the coding sequences of active genes. On the other hand, DNA repair at inactive genes or silent areas of the genome is not regulated by Rad26p. However, how Rad26p recognizes DNA lesions at the actively transcribing genes to facilitate DNA repair is not clearly understood in vivo. Here, we show that Rad26p associates with the coding sequences of genes in a transcription-dependent manner, but independently of DNA lesions induced by 4-nitroquinoline-1-oxide in Saccharomyces cerevisiae. Further, histone H3 lysine 36 methylation that occurs at the active coding sequence stimulates the recruitment of Rad26p. Intriguingly, we find that Rad26p is recruited to the site of DNA lesion in an elongating RNA polymerase II-dependent manner. However, Rad26p does not recognize DNA lesions in the absence of active transcription. Together, these results provide an important insight as to how Rad26p is delivered to the damage sites at the active, but not inactive, genes to stimulate repair in vivo, shedding much light on the early steps of transcription coupled repair in living eukaryotic cells. PMID- 20007605 TI - Involvement of Vts1, a structure-specific RNA-binding protein, in Okazaki fragment processing in yeast. AB - The non-essential VTS1 gene of Saccharomyces cerevisiae is highly conserved in eukaryotes and encodes a sequence- and structure-specific RNA-binding protein. The Vts1 protein has been implicated in post-transcriptional regulation of a specific set of mRNAs that contains its-binding site at their 3'-untranslated region. In this study, we identified VTS1 as a multi-copy suppressor of dna2 K1080E, a lethal mutant allele of DNA2 that lacks DNA helicase activity. The suppression was allele-specific, since overexpression of Vts1 did not suppress the temperature-dependent growth defects of dna2Delta405N devoid of the N terminal 405-amino-acid residues. Purified recombinant Vts1 stimulated the endonuclease activity of wild-type Dna2, but not the endonuclease activity of Dna2Delta405N, indicating that the activation requires the N-terminal domain of Dna2. Stimulation of Dna2 endonuclease activity by Vts1 appeared to be the direct cause of suppression, since the multi-copy expression of Dna2-K1080E suppressed the lethality observed with its single-copy expression. We found that vts1Delta dna2Delta405N and vts1Deltadna2-7 double mutant cells displayed synergistic growth defects, in support of a functional interaction between two genes. Our results provide both in vivo and in vitro evidence that Vts1 is involved in lagging strand synthesis by modulating the Dna2 endonuclease activity that plays an essential role in Okazaki fragment processing. PMID- 20007606 TI - Insights into the structure, function and evolution of the radical-SAM 23S rRNA methyltransferase Cfr that confers antibiotic resistance in bacteria. AB - The Cfr methyltransferase confers combined resistance to five classes of antibiotics that bind to the peptidyl tranferase center of bacterial ribosomes by catalyzing methylation of the C-8 position of 23S rRNA nucleotide A2503. The same nucleotide is targeted by the housekeeping methyltransferase RlmN that methylates the C-2 position. Database searches with the Cfr sequence have revealed a large group of closely related sequences from all domains of life that contain the conserved CX(3)CX(2)C motif characteristic of radical S-adenosyl-l-methionine (SAM) enzymes. Phylogenetic analysis of the Cfr/RlmN family suggests that the RlmN subfamily is likely the ancestral form, whereas the Cfr subfamily arose via duplication and horizontal gene transfer. A structural model of Cfr has been calculated and used as a guide for alanine mutagenesis studies that corroborate the model-based predictions of a 4Fe-4S cluster, a SAM molecule coordinated to the iron-sulfur cluster (SAM1) and a SAM molecule that is the putative methyl group donor (SAM2). All mutations at predicted functional sites affect Cfr activity significantly as assayed by antibiotic susceptibility testing and primer extension analysis. The investigation has identified essential amino acids and Cfr variants with altered reaction mechanisms and represents a first step towards understanding the structural basis of Cfr activity. PMID- 20007607 TI - Membrane association of mitochondrial DNA facilitates base excision repair in mammalian mitochondria. AB - Mitochondrial DNA encodes a set of 13 polypeptides and is subjected to constant oxidative stress due to ROS production within the organelle. It has been shown that DNA repair in the mitochondrion proceeds through both short- and long-patch base excision repair (BER). In the present article, we have used the natural competence of mammalian mitochondria to import DNA and study the sub mitochondrial localization of the repair system in organello. Results demonstrate that sequences corresponding to the mtDNA non-coding region interact with the inner membrane in a rapid and saturable fashion. We show that uracil containing import substrates are taken into the mitochondrion and are used as templates for damage driven DNA synthesis. After further sub-fractionation, we show that the length of the repair synthesis patch differs in the soluble and the particulate fraction. Bona fide long patch BER synthesis occurs on the DNA associated with the particulate fraction, whereas a nick driven DNA synthesis occurs when the uracil containing DNA accesses the soluble fraction. Our results suggest that coordinate interactions of the different partners needed for BER is only found at sites where the DNA is associated with the membrane. PMID- 20007608 TI - Crystal structure and substrate specificity of plant adenylate isopentenyltransferase from Humulus lupulus: distinctive binding affinity for purine and pyrimidine nucleotides. AB - Cytokinins are important plant hormones, and their biosynthesis most begins with the transfer of isopentenyl group from dimethylallyl diphosphate (DMAPP) to the N6-amino group of adenine by either adenylate isopentenyltransferase (AIPT) or tRNA-IPT. Plant AIPTs use ATP/ADP as an isopentenyl acceptor and bacterial AIPTs prefer AMP, whereas tRNA-IPTs act on specific sites of tRNA. Here, we present the crystal structure of an AIPT-ATP complex from Humulus lupulus (HlAIPT), which is similar to the previous structures of Agrobacterium AIPT and yeast tRNA-IPT. The enzyme is structurally homologous to the NTP-binding kinase family of proteins but forms a solvent-accessible channel that binds to the donor substrate DMAPP, which is directed toward the acceptor substrate ATP/ADP. When measured with isothermal titration calorimetry, some nucleotides displayed different binding affinities to HlAIPT with an order of ATP > dATP approximately ADP > GTP > CTP > UTP. Two basic residues Lys275 and Lys220 in HlAIPT interact with the beta and gamma-phosphate of ATP. By contrast, the interactions are absent in Agrobacterium AIPT because they are replaced by the acidic residues Asp221 and Asp171. Despite its structural similarity to the yeast tRNA-IPT, HlAIPT has evolved with a different binding strategy for adenylate. PMID- 20007610 TI - Myocarditis in a juvenile patient with influenza A virus infection. PMID- 20007611 TI - Rare angiographic and echocardiographic findings of an aortic arch interruption in a patient with differential cyanosis. PMID- 20007612 TI - Spontaneous closure of a coronary fistula due to cardiac allograft vasculopathy. PMID- 20007613 TI - The effect of nutritionally induced hyperlipidaemia on in vitro bovine embryo quality. AB - BACKGROUND: Obesity is associated with female reproductive abnormalities. Hyperlipidaemia might alter the embryonic micro-environment and potentially result in reduced fertility. We aimed to induce hyperlipidaemia nutritionally and investigate the consequences of hyperlipidaemic culture conditions on bovine in vitro embryo development, embryo quality and gene expression patterns. METHODS: Bovine zygotes (n = 1545) were cultured in synthetic oviductal fluid medium supplemented with serum from heifers (n = 3), each fed three successive dietary treatments: (i) control serum, following a hay-based diet, (ii) hyperlipidaemic serum, following a carbohydrate and protected palm-oil-rich diet (FatCh) or (iii) hyperlipidaemic serum, following a protected palm-oil-rich diet (Fat). Blastocysts were evaluated for development, cell count, picnotic and mitotic indexes and cryotolerance. Selected mRNA transcripts were measured by quantitative RT-PCR. RESULTS: FatCh and Fat diets approximately doubled the total cholesterol concentrations, compared with controls (167.1 +/- 11.9, 150.0 +/- 12.8 versus 83.4 +/- 13.7 mg/dl, respectively, P < 0.05), and fatty acid concentrations (8146.60 +/- 214.61, 6935.56 +/- 1081.04 versus 3944.0 +/- 425.07 micromol/l, respectively, P < 0.05). Supplementation of culture media with FatCh and Fat serum significantly reduced blastocyst rates, compared with controls (27.8, 23.4% versus 36.2%, respectively, P < 0.01), total cell number (103.3 +/- 30.1, 95.6 +/- 28.2 versus 146.9 +/- 34.2, respectively, P < 0.01), mitotic index (1.3 +/- 1.1, 1.7 +/- 2.4 versus 3.6 +/- 2.2%, respectively, P < 0.01) and hatching rates after vitrification (20.4, 13.8 versus 35.7%, respectively, P = 0.03). Embryos in FatCh and Fat groups exhibited significantly higher mRNA levels for genes related to apoptosis and metabolism, compared with controls. CONCLUSIONS: This combined in vivo and in vitro model indicates that the exposure of preimplantation embryos to hyperlipidaemic conditions may result in reduced embryo quality and developmental potential, possibly resulting in poorer fertility. PMID- 20007614 TI - Analysis of matrix metalloproteinase-7 expression in eutopic and ectopic endometrium samples from patients with different forms of endometriosis. AB - BACKGROUND: The objective of the present study was to expand our understanding of the role of matrix metalloproteinase-7 (MMP-7) in the pathophysiology of endometriosis. METHODS: Expression levels of MMP-7 mRNA and protein in the eutopic endometrium and ectopic endometrium of patients with different forms of endometriosis were measured with immunohistochemistry and real-time RT-PCR. Endometrial tissues from patients with uterine myomas and those with macroscopically normal pelvic cavities were included as comparison groups. The real-time RT-PCR utilized endometrial cells isolated by laser capture microdissection. MMP-7 immunostained cells were quantified using a computerized image analysis system. RESULTS: MMP-7 expression levels were significantly higher in the endometrial epithelial cells from patients with deep infiltrating endometriosis compared with those isolated from the endometria of patients with only superficial peritoneal endometriosis, uterine myomas or normal endometrium, in the proliferative, late secretory and menstrual phases. MMP-7 protein expression was detected in the ectopic endometrial epithelial cells of 13 samples of deep infiltrating endometriosis (24.5%), 11 samples of ovarian endometriosis (28.6%), 23 samples of black peritoneal lesions (76.7%) and 24 samples of red peritoneal lesions (100%). MMP-7 protein expression in epithelial cells was significantly higher in red peritoneal lesions compared with that of deep infiltrating endometriosis, ovarian endometriosis and black peritoneal lesions, in all phases of the menstrual cycle. CONCLUSION: These findings suggest that MMP 7 expression levels vary significantly among the different forms of endometriosis. PMID- 20007615 TI - A plea for data on the sexes of human offspring and the time of formation of the zygotes within the fruitful cycle. PMID- 20007616 TI - A fatal case of ovarian hyperstimulation syndrome with perforated duodenal ulcer. PMID- 20007618 TI - Central venous saturation: a prognostic tool in cardiac surgery patients. AB - BACKGROUND: Central venous oxygen saturation (ScvO(2)) is a valuable prognostic marker in sepsis. However, its value in cardiac surgery has not been assessed yet. This study aimed at evaluating ScvO(2) as a tool for predicting short-term organ dysfunction (OD) after cardiac surgery. METHODS: A prospective cohort including cardiac surgery patients submitted to a goal-oriented therapy to maintain ScvO(2) above 70% was studied. Postoperative blood samples collected at 30 minutes (T1), 6 hours (T2), and 24 hours (T3) for ScvO(2) measurement were selected to further analysis. Two groups were formed according to the absence (G0) or presence (G1) of OD defined as a Sequential Organ Failure Assessment (SOFA) score >or=5 on the third postoperative day. A logistic regression analysis was performed to identify the variables independently associated with OD on the third postoperative day. RESULTS: From the 246 patients included, 54 (22%) developed OD and were defined as G1. The mortality rates in G0 and G1 were 1.6% and 31.5%, respectively (P < .001). In the comparative analysis between G0 and G1, the ScvO(2) values were remarkably lower in G1 at T1 (66.2 +/- 9.2 vs 62.3 +/ 11.6; P = .009), T2 (69.6 +/- 5.9 vs 63.5 +/- 9.4; P 14 days in a hospital in the last month of life. However, they were significantly less likely than patients of other physician specialists to visit the emergency room (ER) more than once and to use intensive care unit (ICU) care, cardiopulmonary resuscitation (CPR), intubation, and mechanical ventilation in the last month of life. CONCLUSION: The quality of EOL cancer care in Taiwan varied significantly by physician specialty. Cancer decedents cared for by medical oncologists were more likely to receive chemotherapy and prolonged hospitalization but less likely to have multiple ER visits, ICU care, or undergo CPR, intubation, or mechanical ventilation in the last month of life than patients of other physician specialists. PMID- 20007641 TI - Surface area assessment of the murine intestinal tract as a prerequisite for oral dose translation from mouse to man. AB - In many pharmacological and toxicological studies knowledge about the intestinal absorption, which is dependent upon the surface area of absorptive epithelia, is indispensible. Although mice are often used in such preclinical studies, very few quantitative data about their intestinal surface area are available. Especially for locally acting candidate drugs in development, this information is crucial for dose translation towards humans. Therefore, the surface area of the intestinal tract of CD-1 IGS mice was assessed in the present study. The intestinal tracts of 12 mice were collected after euthanasia. From six animals, histological sections from the duodenum, jejunum, ileum, caecum and colon-rectum were made according to common stereological principles. Using these sections, the volumes and surface areas of each intestinal segment were estimated applying stereological counting procedures. In the other six animals, the density and surface area of the microvilli present in each intestinal segment were determined by means of scanning and transmission electron microscopy to assess the increase of the intestinal surface area attributable to the presence of microvilli. The mean total volume and surface area of the intestinal tract were 1.34 cm(3) and 1.41 m(2), respectively. The relative intestinal surface area (intestinal surface area divided by the body surface area) was 119. The relative intestinal surface area of mice is very similar to that of humans. The results of this study are important for the appropriate dose translation of candidate therapeutic compounds in drug development from mouse to humans. PMID- 20007643 TI - The case for chemotherapy only for localized Hodgkin lymphoma. AB - Although radiation therapy (RT) alone, and more recently in combination with chemotherapy, has been the cornerstone of curative treatment for early-stage Hodgkin lymphoma for >40 years, the late toxicities of this treatment are increasingly being recognized as causes of significant late morbidity and mortality. In recent years, the focus of treatment of early-stage Hodgkin lymphoma has been reductions in the doses and fields of RT and attempts to reduce the amount and the toxicity of chemotherapy. Another approach has been to treat with chemotherapy alone and to eliminate RT entirely unless it is absolutely necessary. This review summarizes the accumulating information on the late side effects of treatment of Hodgkin lymphoma, the results of trials with chemotherapy only, attempts to reduce the toxicity of chemotherapy, and the new role of positron emission tomography in treatment planning. PMID- 20007644 TI - Putting insulin glargine and malignancies into perspective. PMID- 20007645 TI - Mysterious metformin. PMID- 20007646 TI - When "flawed" translates into "flood": the unproven association between cancer incidence and glargine insulin therapy. PMID- 20007647 TI - Superoxide radical induces sclerotial differentiation in filamentous phytopathogenic fungi: a superoxide dismutase mimetics study. AB - This study shows that the superoxide radical (O(2) *( -)), a direct indicator of oxidative stress, is involved in the differentiation of the phytopathogenic filamentous fungi Rhizoctonia solani, Sclerotinia sclerotiorum, Sclerotium rolfsii and Sclerotinia minor, shown by using superoxide dismutase (SOD) mimetics to decrease their sclerotial differentiation. The production rate of O(2) *(-) and SOD levels in these fungi, as expected, were significantly lowered by the SOD mimetics, with concomitant decrease of the indirect indicator of oxidative stress, lipid peroxidation. PMID- 20007648 TI - Racemase activity effected by two dehydrogenases in sulfolactate degradation by Chromohalobacter salexigens: purification of (S)-sulfolactate dehydrogenase. AB - Chromohalobacter salexigens DSM 3043, whose genome has been sequenced, is known to degrade (R,S)-sulfolactate as a sole carbon and energy source for growth. Utilization of the compound(s) was shown to be quantitative, and an eight-gene cluster (Csal_1764-Csal_1771) was hypothesized to encode the enzymes in the degradative pathway. It comprised a transcriptional regulator (SuyR), a Tripartite Tricarboxylate Transporter-family uptake system for sulfolactate (SlcHFG), two sulfolactate dehydrogenases of opposite sulfonate stereochemistry, namely novel SlcC and ComC [(R)-sulfolactate dehydrogenase] [EC 1.1.1.272] and desulfonative sulfolactate sulfo-lyase (SuyAB) [EC 4.4.1.24]. Inducible reduction of 3-sulfopyruvate, inducible SuyAB activity and induction of an unknown protein were detected. Separation of the soluble proteins from induced cells on an anion exchange column yielded four relevant fractions. Two different fractions reduced sulfopyruvate with NAD(P)H, a third yielded SuyAB activity, and the fourth contained the unknown protein. The latter was identified by peptide-mass fingerprinting as SlcH, the candidate periplasmic binding protein of the transport system. Separated SuyB was also identified by peptide-mass fingerprinting. ComC was partially purified and identified by peptide-mass fingerprinting. The (R)-sulfolactate that ComC produced from sulfopyruvate was a substrate for SuyAB, which showed that SuyAB is (R)-sulfolactate sulfo-lyase. SlcC was purified to homogeneity. This enzyme also formed sulfolactate from sulfopyruvate, but the latter enantiomer was not a substrate for SuyAB. SlcC was obviously ( S)-sulfolactate dehydrogenase. PMID- 20007649 TI - Direct interaction of iron-regulated surface determinant IsdB of Staphylococcus aureus with the GPIIb/IIIa receptor on platelets. AB - The interaction of bacteria with platelets is implicated in the pathogenesis of endovascular infections, including infective endocarditis, of which Staphylococcus aureus is the leading cause. Several S. aureus surface proteins mediate aggregation of platelets by fibrinogen- or fibronectin-dependent processes, which also requires specific antibodies. In this study S. aureus was grown in iron-limited medium to mimic in vivo conditions in which iron is unavailable to pathogens. Under such conditions, a S. aureus mutant lacking the known platelet-activating surface proteins adhered directly to platelets in the absence of plasma proteins and triggered aggregation. Platelet adhesion and aggregation was prevented by inhibiting expression of iron-regulated surface determinant (Isd) proteins. Mutants defective in IsdB, but not IsdA or IsdH, were unable to adhere to or aggregate platelets. Antibodies to the platelet integrin GPIIb/IIIa inhibited platelet adhesion by IsdB-expressing strains, as did antagonists of GPIIb/IIIa. Surface plasmon resonance demonstrated that recombinant IsdB interacts directly with GPIIb/IIIa. PMID- 20007650 TI - Treponema denticola biofilm-induced expression of a bacteriophage, toxin antitoxin systems and transposases. AB - Treponema denticola is an oral spirochaete that has been strongly associated with chronic periodontitis. The bacterium exists as part of a dense biofilm (subgingival dental plaque) accreted to the tooth. To determine T. denticola gene products important for persistence as a biofilm we developed a continuous-culture biofilm model and conducted a genome-wide transcriptomic analysis of biofilm and planktonic cells. A total of 126 genes were differentially expressed with a fold change of 1.5 or greater. This analysis identified the upregulation of putative prophage genes in the T. denticola 35405 genome. Intact bacteriophage particles were isolated from T. denticola and circular phage DNA was detected by PCR analysis. This represents the first, to our knowledge, functional bacteriophage isolated from T. denticola, which we have designated varphitd1. In biofilm cells there was also an upregulation of genes encoding several virulence factors, toxin antitoxin systems and a family of putative transposases. Together, these data indicate that there is a higher potential for genetic mobility in T. denticola when growing as a biofilm and that these systems are important for the biofilm persistence and therefore virulence of this bacterium. PMID- 20007651 TI - Regulation of dsr genes encoding proteins responsible for the oxidation of stored sulfur in Allochromatium vinosum. AB - Sulfur globules are formed as obligatory intermediates during the oxidation of reduced sulfur compounds in many environmentally important photo- and chemolithoautotrophic bacteria. It is well established that the so-called Dsr proteins are essential for the oxidation of zero-valent sulfur accumulated in the globules; however, hardly anything is known about the regulation of dsr gene expression. Here, we present a closer look at the regulation of the dsr genes in the phototrophic sulfur bacterium Allochromatium vinosum. The dsr genes are expressed in a reduced sulfur compound-dependent manner and neither sulfite, the product of the reverse-acting dissimilatory sulfite reductase DsrAB, nor the alternative electron donor malate inhibit the gene expression. Moreover, we show the oxidation of sulfur to sulfite to be the rate-limiting step in the oxidation of sulfur to sulfate as sulfate production starts concomitantly with the upregulation of the expression of the dsr genes. Real-time RT-PCR experiments suggest that the genes dsrC and dsrS are additionally expressed from secondary internal promoters, pointing to a special function of the encoded proteins. Earlier structural analyses indicated the presence of a helix-turn-helix (HTH) like motif in DsrC. We therefore assessed the DNA-binding capability of the protein and provide evidence for a possible regulatory function of DsrC. PMID- 20007652 TI - Blogging as 'therapy'? Exploring personal technologies for smoking cessation. AB - This article presents some early, design-oriented research findings from a study that introduced mobile blogging technologies to four people who wished to make a health-related life change--giving up smoking. We wanted to establish the nature of the relationship between blogging and quitting smoking (if any), inspired by some earlier work in the domain showing that social technologies may help with the quit process. We present an account of three participants, documenting details of how blogging technologies fitted into their (changing) lives and examples of digital content they produced. We describe, using examples from participant blogs, instances of self-expression, replacement and self-awareness. We suggest, despite all participants failing in their quit attempts, that there are possible provisional, therapeutic characteristics to such social technologies. Finally, we suggest this therapeutic process can be understood better through a concept of personal translucence. PMID- 20007653 TI - Prospective tracking of a pediatric emergency department e-kiosk to deliver asthma education. AB - The study objective was to describe the prospective use of an interactive Patient Education and Motivation Tool (PEMT) placed within a pediatric emergency department (ED). A touch screen computer was utilized to deliver asthma education to children and their parents/guardians during their acute asthma visit between November 2006 and April 2007. Ninety-nine participants were enrolled in this prospective non-randomized pre-post study. PEMT comprised three key components: screening, learning and evaluation. The tool tracked the date the system was used, user characteristics, asthma knowledge, amount of time spent on each screen, and navigational patterns of individuals using the program. The results showed that baseline asthma knowledge had positive association with age and negative association with time spent in the learning module. There was negative association between age and time spent in the learning module. Thus PEMT was effective in improving the asthma knowledge of young patients and those having lower baseline knowledge. PMID- 20007654 TI - Development, implementation and evaluation of an electronic medical record prompt for bone density testing. AB - The purpose of this study was to assess whether an electronic prompt promoting BMD testing affected the proportion of patients who received BMD and/or bone health medication. Rheumatology providers of patients 40+, on prednisone, with no record of BMD testing in the past 2 years, were targeted with the message: 'This patient is at risk for osteoporosis due to prednisone use. We have no record of a recent Dexa scan.' We also surveyed providers on the prompt's value. The use of prednisone remained stable; BMD testing was quite low in all periods but increased slightly (non-statistically) over time, as did bone health medication use. Providers found the prompt not tailored enough to be clinically meaningful. Electronic prompts seem beneficial in theory; however, putting them into practice has challenges. While the EMR has great potential to improve care, more needs to be done to ensure optimal use. PMID- 20007655 TI - Data extraction from a semi-structured electronic medical record system for outpatients: a model to facilitate the access and use of data for quality control and research. AB - The use of clinical data from electronic medical records (EMRs) for clinical research and for evaluation of quality of care requires an extraction process. Many efforts have failed because the extracted data seemed to be unstructured, incomplete and ridden by errors. We have developed and tested a concept of extracting semi-structured EMRs (Journal III, Profdoc) data from 776 diabetes patients in a general practice clinic over a 5 year period. We used standard database management techniques commonly applied in clinical research in the pharmaceutical industry to clean up the data and make the data available for statistical analysis. The key problem was difficulties locating the data, as no standard way to enter the data in the EMR system was reinforced. Furthermore, no built-in edit checks to facilitate data entry were available. Laboratory, drug information and diagnostic data could be used directly while other data such as vital signs required much work to locate and become useful. PMID- 20007656 TI - A woman with acute myelopathy in pregnancy: case outcome. PMID- 20007657 TI - Commentary: Managing clinicians' assessment. PMID- 20007658 TI - Commentary: Patient's view. PMID- 20007659 TI - Commentary: Complex medical conditions in pregnancy need appropriate multidisciplinary input. PMID- 20007660 TI - Commentary: Learning out of your depth. PMID- 20007661 TI - Commentary: Neurological perspective. PMID- 20007663 TI - Medicine in the News. Health stories of the decade. PMID- 20007664 TI - Sit-to-stand movement as a performance-based measure for patients with total knee arthroplasty. AB - BACKGROUND: Functional recovery of patients after a total knee arthroplasty (TKA) usually is measured with questionnaires. However, these self-report measures assess the patient's perspective on his or her ability to perform a task. Performance-based tests are needed to assess the patient's actual ability to perform a task. OBJECTIVE: The main purpose of this study was to quantify improvement in performance of the sit-to-stand movement of patients with a TKA. DESIGN AND METHODS: In this prospective study of 16 patients with end-stage knee osteoarthritis followed by a TKA, the maximal knee angular extension velocity and amount of unloading (shifting weight) of the affected leg during the sit-to-stand movement and the visual analog scale score for pain were assessed preoperatively and 6 months and 1 year postoperatively. These data were compared with data for a control group of individuals who were healthy (n=27). RESULTS: Before surgery, the participants in the TKA group unloaded their affected leg, but within 6 months after implantation, the affected leg was almost fully loaded again and comparable to the loading symmetry ratio of the control group. Furthermore, knee extension velocity also had increased, but remained lower than that of the control group. The changes in knee extension velocity took place during the first 6 months, after which a plateau was visible. Limitations A potential limitation of the study design was that the patients were not perfectly matched with the control subjects. CONCLUSIONS: Implantation of a total knee prosthesis partly improved performance of the sit-to-stand movement. Participants in the TKA group could fully load their operated leg, but they could not generate enough knee angular velocity during rising compared with the control group. PMID- 20007665 TI - Mortality from pandemic A/H1N1 2009 influenza in England: public health surveillance study. AB - OBJECTIVE: To establish mortality from pandemic A/H1N1 2009 influenza up to 8 November 2009. DESIGN: Investigation of all reported deaths related to pandemic A/H1N1 in England. SETTING: Mandatory reporting systems established in acute hospitals and primary care. PARTICIPANTS: Physicians responsible for the patient. MAIN OUTCOME MEASURES: Numbers of deaths from influenza combined with mid-range estimates of numbers of cases of influenza to calculate age specific case fatality rates. Underlying conditions, time course of illness, and antiviral treatment. RESULTS: With the official mid-range estimate for incidence of pandemic A/H1N1, the overall estimated case fatality rate was 26 (range 11-66) per 100 000. It was lowest for children aged 5-14 (11 (range 3-36) per 100 000) and highest for those aged >or=65 (980 (range 300-3200) per 100 000). In the 138 people in whom the confirmed cause of death was pandemic A/H1N1, the median age was 39 (interquartile range 17-57). Two thirds of patients who died (92, 67%) would now be eligible for the first phase of vaccination in England. Fifty (36%) had no, or only mild, pre-existing illness. Most patients (108, 78%) had been prescribed antiviral drugs, but of these, 82 (76%) did not receive them within the first 48 hours of illness. CONCLUSIONS: Viewed statistically, mortality in this pandemic compares favourably with 20th century influenza pandemics. A lower population impact than previous pandemics, however, is not a justification for public health inaction. Our data support the priority vaccination of high risk groups. We observed delayed antiviral use in most fatal cases, which suggests an opportunity to reduce deaths by making timely antiviral treatment available, although the lack of a control group limits the ability to extrapolate from this observation. Given that a substantial minority of deaths occur in previously healthy people, there is a case for extending the vaccination programme and for continuing to make early antiviral treatment widely available. PMID- 20007666 TI - Effect of 20 mph traffic speed zones on road injuries in London, 1986-2006: controlled interrupted time series analysis. AB - OBJECTIVE: To quantify the effect of the introduction of 20 mph (32 km an hour) traffic speed zones on road collisions, injuries, and fatalities in London. DESIGN: Observational study based on analysis of geographically coded police data on road casualties, 1986-2006. Analyses were made of longitudinal changes in counts of road injuries within each of 119 029 road segments with at least one casualty with conditional fixed effects Poisson models. Estimates of the effect of introducing 20 mph zones on casualties within those zones and in adjacent areas were adjusted for the underlying downward trend in traffic casualties. SETTING: London. MAIN OUTCOME MEASURES: All casualties from road collisions; those killed and seriously injured (KSI). RESULTS: The introduction of 20 mph zones was associated with a 41.9% (95% confidence interval 36.0% to 47.8%) reduction in road casualties, after adjustment for underlying time trends. The percentage reduction was greatest in younger children and greater for the category of killed or seriously injured casualties than for minor injuries. There was no evidence of casualty migration to areas adjacent to 20 mph zones, where casualties also fell slightly by an average of 8.0% (4.4% to 11.5%). Conclusions 20 mph zones are effective measures for reducing road injuries and deaths. PMID- 20007667 TI - Traffic speed zones and road injuries. PMID- 20007668 TI - Neglected tropical diseases. AB - INTRODUCTION: The neglected tropical diseases (NTDs) are infectious diseases that principally impact the world's poorest people. They have been neglected for decades, initially as part of a general disregard for the developing world, and more recently due to the intensity of focus on HIV/AIDS, tuberculosis and malaria. SOURCES OF DATA: Primary research and review articles were selected for inclusion using searches of PubMed and our existing collections. RESULTS: There have been recent notable successes in NTD control. Dracunculiasis is approaching eradication. Leprosy and onchocerciasis are in decline. There are ambitious plans to eliminate trachoma and lymphatic filariasis. Investment in NTD control has high rates of economic return. CONCLUSION: Although there are proven strategies to control several NTDs, these diseases continue to cause a massive burden of morbidity. There is urgent need for more basic and operational research, drug and vaccine development, and greater prioritization by governments and international agencies. PMID- 20007669 TI - Glucuronidation of psilocin and 4-hydroxyindole by the human UDP glucuronosyltransferases. AB - We have examined the glucuronidation of psilocin, a hallucinogenic indole alkaloid, by the 19 recombinant human UDP-glucuronosyltransferases (UGTs) of subfamilies 1A, 2A, and 2B. The glucuronidation of 4-hydroxyindole, a related indole that lacks the N,N-dimethylaminoethyl side chain, was studied as well. UGT1A10 exhibited the highest psilocin glucuronidation activity, whereas the activities of UGTs 1A9, 1A8, 1A7, and 1A6 were significantly lower. On the other hand, UGT1A6 was by far the most active enzyme mediating 4-hydroxyindole glucuronidation, whereas the activities of UGTs 1A7-1A10 toward 4-hydroxyindole resembled their respective psilocin glucuronidation rates. Psilocin glucuronidation by UGT1A10 followed Michaelis-Menten kinetics in which psilocin is a low-affinity high-turnover substrate (K(m) = 3.8 mM; V(max) = 2.5 nmol/min/mg). The kinetics of psilocin glucuronidation by UGT1A9 was more complex and may be best described by biphasic kinetics with both intermediate (K(m1) = 1.0 mM) and very low affinity components. The glucuronidation of 4-hydroxyindole by UGT1A6 exhibited higher affinity (K(m) = 178 microM) and strong substrate inhibition. Experiments with human liver and intestinal microsomes (HLM and HIM, respectively) revealed similar psilocin glucuronidation activity in both samples, but a much higher 4-hydroxyindole glucuronidation rate was found in HLM versus HIM. The expression levels of UGTs 1A6-1A10 in different tissues were studied by quantitative real-time-PCR, and the results, together with the activity assays findings, suggest that whereas psilocin may be subjected to extensive glucuronidation by UGT1A10 in the small intestine, UGT1A9 is likely the main contributor to its glucuronidation once it has been absorbed into the circulation. PMID- 20007670 TI - Identification of cytochrome P450 isoforms involved in the metabolism of paroxetine and estimation of their importance for human paroxetine metabolism using a population-based simulator. AB - We identify here for the first time the low-affinity cytochrome P450 (P450) isoforms that metabolize paroxetine, using cDNA-expressed human P450s measuring substrate depletion and paroxetine-catechol (product) formation by liquid chromatography-tandem mass spectrometry. CYP1A2, CYP2C19, CYP2D6, CYP3A4, and CYP3A5 were identified as paroxetine-catechol-forming P450 isoforms, and CYP2C19 and CYP2D6 were identified as metabolizing P450 isoforms by substrate depletion. Michaelis-Menten constants K(m) and V(max) were determined by product formation and substrate depletion. Using selective inhibitory studies and a relative activity factor approach for pooled and single-donor human liver microsomes, we confirmed involvement of the identified P450 isoforms for paroxetine-catechol formation at 1 and 20 muM paroxetine. In addition, we used the population-based simulator Simcyp to estimate the importance of the identified paroxetine metabolizing P450 isoforms for human metabolism, taking mechanism-based inhibition into account. The amount of active hepatic CYP2D6 and CYP3A4 (not inactivated by mechanism-based inhibition) was also estimated by Simcyp. For extensive and poor metabolizers of CYP2D6, Simcyp-estimated pharmacokinetic profiles were in good agreement with those reported in published in vivo studies. Considering the kinetic parameters, inhibition results, relative activity factor calculations, and Simcyp simulations, CYP2D6 (high affinity) and CYP3A4 (low affinity) are most likely to be the major contributors to paroxetine metabolism in humans. For some individuals CYP1A2 could be of importance for paroxetine metabolism, whereas the importance of CYP2C19 and CYP3A5 is probably limited. PMID- 20007672 TI - Distributed structures underlie gating differences between the kin channel KAT1 and the Kout channel SKOR. AB - The family of voltage-gated (Shaker-like) potassium channels in plants includes both inward-rectifying (K(in)) channels that allow plant cells to accumulate K(+) and outward-rectifying (K(out)) channels that mediate K(+) efflux. Despite their close structural similarities, K(in) and K(out) channels differ in their gating sensitivity towards voltage and the extracellular K(+) concentration. We have carried out a systematic program of domain swapping between the K(out) channel SKOR and the K(in) channel KAT1 to examine the impacts on gating of the pore regions, the S4, S5, and the S6 helices. We found that, in particular, the N terminal part of the S5 played a critical role in KAT1 and SKOR gating. Our findings were supported by molecular dynamics of KAT1 and SKOR homology models. In silico analysis revealed that during channel opening and closing, displacement of certain residues, especially in the S5 and S6 segments, is more pronounced in KAT1 than in SKOR. From our analysis of the S4-S6 region, we conclude that gating (and K(+)-sensing in SKOR) depend on a number of structural elements that are dispersed over this approximately 145-residue sequence and that these place additional constraints on configurational rearrangement of the channels during gating. PMID- 20007673 TI - Impact of improved classification on the association of human papillomavirus with cervical precancer. AB - Misclassification of exposure and surrogate endpoints of disease can obscure causal relations. Using data from the Atypical Squamous Cells of Undetermined Significance/Low-Grade Squamous Intraepithelial Lesion Triage Study (ALTS, 1997 2001), the authors explored the impact of exposure (human papillomavirus (HPV) detection) and endpoint (histologic cervical precancer) classification on their mutual association. Women referred into this study with an atypical squamous cells of undetermined significance Papanicolaou test with satisfactory results for all 4 HPV tests were included in this analysis (n = 3,215; 92.2%). HPV testing results were related to different definitions of cervical precancer, based on paired, worst 2-year histologic diagnoses, by calculating clinical sensitivity, specificity, and odds ratios. The authors found that HPV test sensitivity increased and specificity decreased with increasing certainty of cervical precancer, with HPV testing having the highest sensitivity (92%-98%) and lowest specificity (46%-54%) for consensus cervical intraepithelial neoplasia grade 3 (CIN 3). The overall accuracy of each HPV test, as measured by odds ratios, was greatest for consensus CIN-3 diagnoses, from 2- to 4-fold greater than for a less stringent precancer definition of any diagnosis of CIN 2 or more severe. In summary, there was convergence of greater certainty of carcinogenic HPV with greater certainty of a precancerous diagnosis, such that all 4 HPV tests almost always tested positive in women most likely to have cervical precancer. Finding increasingly strong associations when both test and diagnostic misclassification are reduced is a useful sign of "true association" in molecular epidemiology. PMID- 20007674 TI - Estimates of the transmissibility of the 1968 (Hong Kong) influenza pandemic: evidence of increased transmissibility between successive waves. AB - The transmissibility of the strain of influenza virus which caused the 1968 influenza pandemic is poorly understood. Increases in outbreak size between the first and second waves suggest that it may even have increased between successive waves. The authors estimated basic and effective reproduction numbers for both waves of the 1968 influenza pandemic. Epidemic curves and overall attack rates for the 1968 pandemic, based on clinical and serologic data, were retrieved from published literature. The basic and effective reproduction numbers were estimated from 46 and 17 data sets for the first and second waves, respectively, based on the growth rate and/or final size of the epidemic. Estimates of the basic reproduction number (R(0)) were in the range of 1.06-2.06 for the first wave and, assuming cross-protection, 1.21-3.58 in the second. Within each wave, there was little geographic variation in transmissibility. In the 10 settings for which data were available for both waves, R(0) was estimated to be higher during the second wave than during the first. This might partly explain the larger outbreaks in the second wave as compared with the first. This potential for change in viral behavior may have consequences for future pandemic mitigation strategies. PMID- 20007675 TI - Invited commentary: Human papillomavirus infection and risk of cervical precancer -using the right methods to answer the right questions. AB - Epidemiologists are well aware of the negative consequences of measurement error in exposure and outcome variables to their ability to detect putative causal associations. However, empirical proof that remedying the misclassification problem improves estimates of epidemiologic effect is seldom examined in detail. Of all areas in cancer epidemiology, perhaps the best example of the consequences of misclassification and of the steps taken to circumvent them was the pursuit, beginning in the mid-1980s, of the human papillomavirus (HPV) infection-cervical cancer association. The stakes were high: Had the wrong conclusions been reached epidemiologists would have been led astray in the search for competing hypotheses for the sexually transmissible agent causing cervical cancer or in ascribing to HPV infection a mere ancillary role among many lifestyle, hormonal, and environmental factors. The article by Castle et al. in this issue of the Journal (Am J Epidemiol. 2010;171(2):155-163) provides a detailed account of the joint influences of improved HPV and cervical precancer measurements in gradually unveiling the strong magnitude of the underlying association between viral exposure and cervical lesion risk. In this commentary, the authors extend the findings of Castle et al. by providing additional empirical evidence in support of their arguments. PMID- 20007676 TI - Second chances in mineral metabolism. PMID- 20007678 TI - Novel B cell therapeutic targets in transplantation and immune-mediated glomerular diseases. AB - B cells and antibodies play an important role in the alloresponse to renal grafts as well as in immune-mediated glomerular diseases. In transplantation, greater recognition and improved diagnosis of antibody-mediated rejection have been a catalyst to the introduction of newer drugs and regimens that target B cells, plasma cells, and donor-specific antibodies to improve the outcome associated with antibody-mediated rejection. In immune-mediated renal disease, novel and more selective B cell therapies are gradually modifying the traditional therapeutic approach that consists of steroids and other immunosuppressants. A new era of selective and more effective immunosuppression agents that target the humoral response is finally emerging in transplantation and renal diseases. PMID- 20007677 TI - Urinary netrin-1 is an early predictive biomarker of acute kidney injury after cardiac surgery. AB - BACKGROUND AND OBJECTIVES: Netrin-1, a laminin-related axon guidance molecule, is highly induced and excreted in the urine after acute kidney injury (AKI) in animals. Here, we determined the utility of urinary netrin-1 levels to predict AKI in humans undergoing cardiopulmonary bypass (CPB). DESIGN, SETTING, PARTICIPANTS, & MEASUREMENTS: Serial urine samples were analyzed by enzyme-linked immunosorbent assay for netrin-1 in 26 patients who developed AKI (defined as a 50% or greater increase in serum creatinine after CPB) and 34 controls (patients who did not develop AKI after CPB). RESULTS: Using serum creatinine, AKI was detected on average only 48 hours after CPB. In contrast, urine netrin-1 increased at 2 hours after CPB, peaked at 6 hours (2462 +/- 370 pg/mg creatinine), and remained elevated up to 48 hours after CPB. The predictive power of netrin-1 as demonstrated by area under the receiver-operating characteristics curve for diagnosis of AKI at 2, 6, and 12 hours after CPB was 0.74, 0.86, and 0.89, respectively. The 6-hour urine netrin-1 measurement strongly correlated with duration and severity of AKI, as well as length of hospital stay (all P < 0.05). Adjusting for CPB time, the 6-hour netrin-1 remained a powerful independent predictor of AKI, with an odds ratio of 1.20 (95% confidence interval: 1.08 to 1.41; P = 0.006). CONCLUSION: Our results suggest that netrin-1 is an early, predictive biomarker of AKI after CPB and may allow for the reliable early diagnosis and prognosis of AKI after CPB, before the rise in serum creatinine. PMID- 20007679 TI - Relationship among length of facility ownership, clinical performance, and mortality. AB - BACKGROUND AND OBJECTIVES: The association between level of performance in achieving guideline-recommended clinical indicators and relative reduction in patient mortality is inconsistent among large dialysis organizations (LDOs). Because growth rates among providers differ, we reasoned that clinical performance and mortality rates in dialysis facilities may be related to length of facility ownership. DESIGN, SETTING, PARTICIPANTS, & MEASUREMENTS: We examined achievement of clinical performance indicators among prevalent long-term hemodialysis patients who were enrolled in cohorts of DaVita facilities between December 2005 and December 2007. We compared results in 606 facilities owned before December 1, 2004 (existing), with those seen in 504 facilities that were acquired in October 2005 (newly acquired). RESULTS: At baseline, existing compared with newly acquired DaVita facilities showed higher levels of clinical performance and lower patient mortality. These differences persisted up to 2 years for selected outcomes, including dialysis adequacy and anemia management. Substantial improvement was seen in both cohorts for mineral bone disease outcomes; however, 2 years after acquisition, between-cohort differences in relative risk for death were no longer discernible. CONCLUSIONS: These findings confirm that intervention to improve quality outcomes in dialysis facilities produces direct benefits that are tangible to patients. Our results also provide new evidence that length of ownership may be a significant factor in determining facility performance within a large dialysis organization. PMID- 20007680 TI - Enzyme replacement therapy and Fabry nephropathy. AB - Involvement of the kidneys in Fabry disease ("nephropathy") occurs in male and female individuals. The majority of patients with progressive nephropathy will have significant proteinuria and develop progressive loss of kidney function, leading to ESRD. All too often, treating physicians may ignore "normal" serum creatinine levels or "minimal" proteinuria and fail to assess properly the severity of kidney involvement and institute appropriate management. Fabry nephropathy is treatable, even in patients with fairly advanced disease. Although the cornerstone of therapy remains enzyme replacement therapy with agalsidase, this treatment alone does not reduce urine protein excretion. Treatment with angiotensin receptor blockers or angiotensin-converting enzyme inhibitors must be added to enzyme replacement therapy to reduce urine protein excretion with the hope that this will stabilize kidney function. Kidney function, with at least estimated GFR based on serum creatinine and measurements of urinary protein, should be measured at every clinic visit, and the rate of change of the estimated GFR should be followed over time. Antiproteinuric therapy can be dosed to a prespecified urine protein target rather than a specific BP goal, with the proviso that successful therapy will usually lower the BP below the goal of 130/80 mmHg that is used for other forms of kidney disease. The overall goal for treating Fabry nephropathy is to reduce the rate of loss of GFR to -1 ml/min per 1.73 m(2)/yr, which is that seen in the normal adult population. A systematic approach is presented for reaching this goal in the individual patient. PMID- 20007681 TI - Prevalence of atrial fibrillation and its predictors in nondialysis patients with chronic kidney disease. AB - BACKGROUND AND OBJECTIVES: Chronic kidney disease (CKD) increases systemic inflammation, which is implicated in development and maintenance of atrial fibrillation (AF); therefore, we hypothesized that the prevalence of AF would be increased among nondialysis patients with CKD. This study also reports independent predictors of the presence of AF in this population. DESIGN, SETTING, PARTICIPANTS, & MEASUREMENTS: A retrospective, cross-sectional analysis of 1010 consecutive nondialysis patients with CKD from two community-based hospitals was conducted. Estimated GFRs (eGFRs) were calculated using the Modification of Diet in Renal Disease (MDRD) equation. Multivariate logistic regression was used to determine independent predictors. RESULTS: Of 1010 nondialysis patients with CKD, 214 (21.2%) had AF. Patients with AF were older than patients without AF (76 +/- 11 versus 63 +/- 15 yr). The prevalence of AF among white patients (42.7%) was higher than among black patients (12.7%) or other races (5.7%). In multivariate analyses, age, white race, increasing left atrial diameter, lower systolic BP, and congestive heart failure were identified as independent predictors of the presence of AF. Although serum high-sensitivity C-reactive protein levels were elevated in our population (5.2 +/- 7.4 mg/L), levels did not correlate with the presence of AF or with eGFR. Finally, eGFR did not correlate with the presence of AF in our population. CONCLUSIONS: The prevalence of AF was increased in our population, and independent predictors were age, white race, increasing left atrial diameter, lower systolic BP, and congestive heart failure. PMID- 20007683 TI - Microbial degradation of dimethylsulphide and related C1-sulphur compounds: organisms and pathways controlling fluxes of sulphur in the biosphere. AB - Dimethylsulphide (DMS) plays a major role in the global sulphur cycle. It has important implications for atmospheric chemistry, climate regulation, and sulphur transport from the marine to the atmospheric and terrestrial environments. In addition, DMS acts as an info-chemical for a wide range of organisms ranging from micro-organisms to mammals. Micro-organisms that cycle DMS are widely distributed in a range of environments, for instance, oxic and anoxic marine, freshwater and terrestrial habitats. Despite the importance of DMS that has been unearthed by many studies since the early 1970s, the understanding of the biochemistry, genetics, and ecology of DMS-degrading micro-organisms is still limited. This review examines current knowledge on the microbial cycling of DMS and points out areas for future research that should shed more light on the role of organisms degrading DMS and related compounds in the biosphere. PMID- 20007682 TI - The chloroplast genome sequence of mungbean (Vigna radiata) determined by high throughput pyrosequencing: structural organization and phylogenetic relationships. AB - Mungbean is an economically important crop which is grown principally for its protein-rich dry seeds. However, genomic research of mungbean has lagged behind other species in the Fabaceae family. Here, we reported the complete chloroplast (cp) genome sequence of mungbean obtained by the 454 pyrosequencing technology. The mungbean cp genome is 151 271 bp in length which includes a pair of inverted repeats (IRs) of 26 474 bp separated by a small single-copy region of 17 427 bp and a large single-copy region of 80 896 bp. The genome contains 108 unique genes and 19 of these genes are duplicated in the IR. Of these, 75 are predicted protein-coding genes, 4 ribosomal RNA genes and 29 tRNA genes. Relative to other plant cp genomes, we observed two distinct rearrangements: a 50-kb inversion between accD/rps16 and rbcL/trnK-UUU, and a 78-kb rearrangement between trnH/rpl14 and rps19/rps8. We detected sequence length polymorphism in the cp homopolymeric regions at the intra- and inter-specific levels in the Vigna species. Phylogenetic analysis demonstrated a close relationship between Vigna and Phaseolus in the phaseolinae subtribe and provided a strong support for a monophyletic group of the eurosid I. PMID- 20007684 TI - Identification of flavone phytoalexins and a pathogen-inducible flavone synthase II gene (SbFNSII) in sorghum. AB - Following inoculation with the anthracnose pathogen Colletotrichum sublineolum, seedlings of the sorghum resistant cultivar SC748-5 showed more rapid and elevated accumulation of luteolin than the susceptible cultivar BTx623. On the other hand, apigenin was the major flavone detected in infected BTx623 seedlings. Luteolin was demonstrated to show stronger inhibition of spore germination of C. sublineolum than apigenin. Because of their pathogen-inducible and antifungal nature, both flavone aglycones are considered sorghum phytoalexins. The key enzyme responsible for flavone biosynthesis has not been characterized in monocots. A sorghum pathogen-inducible gene encoding a cytochrome P450 protein (CYP93G3) in the uncharacterized CYP93G subfamily was identified. Transgenic expression of the P450 gene in Arabidopsis demonstrated that the encoded protein is a functional flavone synthase (FNS) II in planta. The sorghum gene was then termed SbFNSII. It is a single-copy gene located on chromosome 2 and the first FNSII gene characterized in a monocot. Metabolite analysis by liquid chromatography-tandem mass spectrometry (LC-MS/MS) in precursor ion scan mode revealed the accumulation of 2-hydroxynaringenin and 2-hydroxyeriodictyol hexosides in the transgenic Arabidopsis plants. Hence, SbFNSII appears to share a similar catalytic mechanism with the licorice and Medicago truncatula FNSIIs (CYP93B subfamily) by converting flavanones to flavone through the formation of 2 hydroxyflavanones. PMID- 20007685 TI - A mutation in GDP-mannose pyrophosphorylase causes conditional hypersensitivity to ammonium, resulting in Arabidopsis root growth inhibition, altered ammonium metabolism, and hormone homeostasis. AB - Ascorbic acid (AA) is an antioxidant fulfilling a multitude of cellular functions. Given its pivotal role in maintaining the rate of cell growth and division in the quiescent centre of the root, it was hypothesized that the AA deficient Arabidopsis thaliana mutants vtc1-1, vtc2-1, vtc3-1, and vtc4-1 have altered root growth. To test this hypothesis, root development was studied in the wild type and vtc mutants grown on Murashige and Skoog medium. It was discovered, however, that only the vtc1-1 mutant has strongly retarded root growth, while the other vtc mutants exhibit a wild-type root phenotype. It is demonstrated that the short-root phenotype in vtc1-1 is independent of AA deficiency and oxidative stress. Instead, vtc1-1 is conditionally hypersensitive to ammonium (NH(4)(+)). To provide new insights into the mechanism of NH(4)(+) sensitivity in vtc1-1, root development, NH(4)(+) content, glutamine synthetase (GS) activity, glutamate dehydrogenase activity, and glutamine content were assessed in wild-type and vtc1 1 mutant plants grown in the presence and absence of high NH(4)(+) and the GS inhibitor MSO. Since VTC1 encodes a GDP-mannose pyrophosphorylase, an enzyme generating GDP-mannose for AA biosynthesis and protein N-glycosylation, it was also tested whether protein N-glycosylation is affected in vtc1-1. Furthermore, since root development requires the action of a variety of hormones, it was investigated whether hormone homeostasis is linked to NH(4)(+) sensitivity in vtc1-1. Our data suggest that NH(4)(+) hypersensitivity in vtc1-1 is caused by disturbed N-glycosylation and that it is associated with auxin and ethylene homeostasis and/or nitric oxide signalling. PMID- 20007686 TI - L-Glutamine inhibits beta-aminobutyric acid-induced stress resistance and priming in Arabidopsis. AB - The non-protein amino acid beta-aminobutyric acid (BABA) enhances Arabidopsis resistance to microbial pathogens and abiotic stresses through potentiation of the Arabidopsis defence responses. In this study, it is shown that BABA induces the stress-induced morphogenic response (SIMR). SIMR is observed in plants exposed to sub-lethal stress conditions. Anthocyanin, a known modulator of stress signalling, was also found to accumulate in BABA-treated Arabidopsis. These data and a previous microarray study indicate that BABA induces a stress response in Arabidopsis. High concentrations of amino acids, except for L-glutamine, cause a general amino acid stress inhibition. General amino acid inhibition is prevented by the addition of L-glutamine. L-Glutamine was found to inhibit the BABA mediated SIMR and anthocyanin accumulation, suggesting that the non-protein amino acid BABA causes a general amino acid stress inhibition in Arabidopsis. L Glutamine also blocked BABA-induced resistance to heat stress and to the virulent bacterial pathogen Pseudomonas syringae pv. tomato DC3000. During bacterial infection, priming of the salicylic acid-dependent defence marker PR1 was abolished by L-glutamine treatment. These results indicate that L-glutamine removal of the BABA-mediated stress response is concomitant with L-glutamine inhibition of BABA priming and BABA-induced resistance. PMID- 20007687 TI - Identification and functional analysis of PCNA1 and PCNA-like1 genes of Phaseolus coccineus. AB - Proliferating cell nuclear antigen (PCNA) is an essential factor in DNA replication and in many other processes in eukaryotic cells. Genetic analysis of Phaseolus coccineus showed the presence of at least two PCNA-like genes in the runner bean genome. Two PCNA genes have previously been found in a few plant species including Arabidopsis, tobacco, and maize. In these species, genes were nearly identical. Two cDNAs of P. coccineus PCNA (PcPCNA1 and PcPCNA-like1) have been identified that differ distinctly from each other. Interestingly, both the genetic organization of PcPCNA1 and PcPCNA-like1 genes and their expression patterns were similar, but these were the only similarities between these genes and their products. The identity between PcPCNA1 and PcPCNA-like1 at the amino acid level was only 54%, with PcPCNA-like1 lacking motifs that are crucial for the activity typical of PCNA. Consequently, these two proteins showed different properties. PcPCNA1 behaved like a typical PCNA protein: it formed a homotrimer and stimulated the activity of human DNA polymerase delta. In addition, PcPCNA1 interacted with a p21 peptide and was recognized by an anti-human PCNA monoclonal antibody PC10. By contrast, PcPCNA-like1 was detected as a monomer and was unable to stimulate the DNA polymerase delta activity. PcPCNA-like1 also could not interact with p21 and was not recognized by the PC10 antibody. Our results suggest that PcPCNA-like1 either is unable to function alone and therefore might be a component of the heterotrimeric PCNA ring or may have other, yet unknown functions. Alternatively, the PcPCNA-like1 gene may represent a pseudogene. PMID- 20007688 TI - High glucose promotes intracellular lipid accumulation in vascular smooth muscle cells by impairing cholesterol influx and efflux balance. AB - AIMS: High glucose promotes macrophage-derived foam cell formation involved in increased influx or reduced efflux of lipids. The aim of this study is to investigate the influence of hyperglycaemia on foam cell transformation of vascular smooth muscle cells (VSMCs) and possible mechanisms contributing to these effects. METHODS AND RESULTS: The results showed that high glucose increased the expression of CD36, a regulator of lipid influx, and suppressed the expression and activity of the adenosine triphosphate-binding cassette (ABC) transporter ABCG1, a regulator of cholesterol efflux to high-density lipoprotein, in a dose- and time-dependent manner. However, cholesterol efflux to lipid-free apoAI was not impaired. VSMCs exposed to high glucose readily developed into lipid-loaded cells, as demonstrated by Oil Red O staining and cholesterol content analysis. In addition, high glucose-induced down-regulation of ABCG1 was reversed by nuclear factor-kappaB (NF-kappaB) inhibitors BAY 11-7085 and tosyl phenylalanine chloromethyl ketone and by the antioxidant N-acetyl-L-cysteine (NAC). This reversal was accompanied by reduced cellular lipid content. Also, NAC and NF-kappaB inhibitors can effectively block the high glucose-induced activity of NF-kappaB binding to DNA and/or peroxide production. CONCLUSION: These results suggested that hyperglycaemia-induced foam cell formation in VSMCs was related to the imbalanced lipid flux by increasing CD36-mediated modified low-density lipoprotein uptake and reducing ABCG1-regulated cellular cholesterol efflux. Moreover, this effect was associated with increased oxidative stress and activated NF-kappaB pathway signalling. PMID- 20007689 TI - Role of HIF-1alpha in proton-mediated CXCR4 down-regulation in endothelial cells. AB - AIMS: Acidification is associated with a variety of pathological and physiological conditions. In the present study, we aimed at investigating whether acidic pH may regulate endothelial cell (EC) functions via the chemokine receptor CXCR4, a key modulator of EC biological activities. METHODS AND RESULTS: Exposure of ECs to acidic pH reversibly inhibited mRNA and protein CXCR4 expression, CXCL12/stromal cell-derived factor (SDF)-1-driven EC chemotaxis in vitro, and CXCR4 expression and activation in vivo in a mouse model. Further, CXCR4 signalling impaired acidosis-induced rescue from apoptosis in ECs. The inhibition of CXCR4 expression occurred transcriptionally and was hypoxia-inducible factor (HIF)-1alpha-dependent as demonstrated by both HIF-1alpha and HIF-1alpha dominant negative overexpression, by HIF-1alpha silencing, and by targeted mutation of the -29 to -25 hypoxia response element (HRE) in the -357/-59 CXCR4 promoter fragment. Moreover, chromatin immunoprecipitation (ChIP) analysis showed endogenous HIF-1alpha binding to the CXCR4 promoter that was enhanced by acidification. CONCLUSION: The results of the present study identify CXCR4 as a key player in the EC response to acidic pH and show, for the first time, that HRE may function not only as an effector of hypoxia, but also as an acidosis response element, and raise the possibility that this may constitute a more general mechanism of transcriptional regulation at acidic pH. PMID- 20007690 TI - MicroRNA-15b modulates cellular ATP levels and degenerates mitochondria via Arl2 in neonatal rat cardiac myocytes. AB - MicroRNAs (miRNAs or miRs) are small, non-coding RNAs that modulate mRNA stability and post-transcriptional translation. A growing body of evidence indicates that specific miRNAs can affect the cellular function of cardiomyocytes. In the present study, miRNAs that are highly expressed in the heart were overexpressed in neonatal rat ventricular myocytes, and cellular ATP levels were assessed. As a result, miR-15b, -16, -195, and -424, which have the same seed sequence, the most critical determinant of miRNA targeting, decreased cellular ATP levels. These results suggest that these miRNAs could specifically down-regulate the same target genes and consequently decrease cellular ATP levels. Through a bioinformatics approach, ADP-ribosylation factor-like 2 (Arl2) was identified as a potential target of miR-15b. It has already been shown that Arl2 localizes to adenine nucleotide transporter 1, the exchanger of ADP/ATP in mitochondria. Overexpression of miR-15b, -16, -195, and -424 suppressed the activity of a luciferase reporter construct fused with the 3'-untranslated region of Arl2. In addition, miR-15b overexpression decreased Arl2 mRNA and protein expression levels. The effects of Arl2 siRNA on cellular ATP levels were the same as those of miR-15b, and the expression of Arl2 could restore ATP levels reduced by miR-15b. A loss-of-function study of miR-15b resulted in increased Arl2 protein and cellular ATP levels. Electron microscopic analysis revealed that mitochondria became degenerated in cardiomyocytes that had been transduced with miR-15b and Arl2 siRNA. The present results suggest that miR-15b may decrease mitochondrial integrity by targeting Arl2 in the heart. PMID- 20007691 TI - Derepression of HMGA2 via removal of ZBRK1/BRCA1/CtIP complex enhances mammary tumorigenesis. AB - The high mobility group AT-hook 2 (HMGA2), a DNA architectural protein, is highly regulated during development and plays an important role in tumorigenesis. Indeed, HMGA2 was overexpressed in many different kinds of tumors. However, the mechanisms regulating HMGA2 expression remain elusive. Using microarray analysis, we found that HMGA2, along with a dozen of other genes, was co-repressed by ZBRK1, BRCA1, and CtIP. BRCA1 exerts its transcriptional repression activity through interaction with the transcriptional repressor ZBRK1 in the central domain, and with CtIP in the C-terminal BRCT domain. Here, we show that ZBRK1, BRCA1, and CtIP form a repression complex that coordinately regulates HMGA2 expression via a ZBRK1 recognition site in the HMGA2 promoter. Depletion of any of the proteins in this complex via adenoviral RNA interference in MCF10A mammary epithelial cells activates HMGA2 expression, resulting in increased colony formation in soft agar. Similarly, depletion of ZBRK1, or ectopic overexpression of HMGA2, in MCF10A cells induces abnormal acinar size with increased cell number and inhibits normal acinar formation. Consistently, many BRCA1-deficient mouse breast tumors express higher levels of HMGA2 than BRCA1-proficient tumors. These results suggest that activation of HMGA2 gene expression through derepression of the ZBRK1/BRCA1/CtIP complex is a significant step in accelerating breast tumorigenesis. PMID- 20007692 TI - Degradation of some polyubiquitinated proteins requires an intrinsic proteasomal binding element in the substrates. AB - Lysine 48-linked polyubiquitin chains usually target proteins for 26 S proteasomal degradation; however, this modification is not a warrant for destruction. Here, we found that efficient degradation of a physiological substrate UbcH10 requires not only an exogenous polyubiquitin chain modification but also its unstructured N-terminal region. Interestingly, the unstructured N terminal region of UbcH10 directly binds the 19 S regulatory complex of the 26 S proteasome, and it mediates the initiation of substrate translocation. To promote ubiquitin-dependent degradation of the folded domains of UbcH10, its N-terminal region can be displaced by exogenous proteasomal binding elements. Moreover, the unstructured N-terminal region can initiate substrate translocation even when UbcH10 is artificially cyclized without a free terminus. Polyubiquitinated circular UbcH10 is completely degraded by the 26 S proteasome. Accordingly, we propose that degradation of some polyubiquitinated proteins requires two binding interactions: a polyubiquitin chain and an intrinsic proteasomal binding element in the substrates (likely an unstructured region); moreover, the intrinsic proteasomal binding element initiates substrate translocation regardless of its location in the substrates. PMID- 20007693 TI - Systematic structure-function analysis of androgen receptor Leu701 mutants explains the properties of the prostate cancer mutant L701H. AB - One mechanism of prostate tumors for escape from androgen ablation therapies is mutation of the androgen receptor (AR). We investigated the unique properties of the AR L701H mutant, which is strongly stimulated by cortisol, by a systematic structure-function analysis. Most amino acid substitutions at position 701 did not affect AR activation by 5alpha-dihydrotestosterone. Further analysis of the AR Leu(701) variants showed that AR L701M and AR L701Q, like AR L701H, had changed ligand responsiveness. AR L701M was strongly activated by progesterone but not by cortisol, whereas the opposite was observed for AR L701Q and AR L701H. Next, we analyzed a panel of structurally related steroids to study which of the OH groups at positions 11beta, 17alpha, and 21, which discriminate cortisol from progesterone, underlie the differential responses to both hormones. The results showed that the 17alpha-OH group was essential for activation of AR L701H and AR L701Q, whereas its absence was important for activation of AR L701M. Modeling indicated a conserved H-bonding network involving the steroidal 17alpha-OH group, His(701) or Gln(701), and the backbone of Ser(778). This network is absent in Leu(701) and in other mutants. A hydrophobic leucine or methionine at position 701 is unfavorable for the 17alpha-OH group. Our results indicate that the specific amino acid residue at position 701, its interaction with the backbone of Ser(778), and the steroidal 17alpha-hydroxyl group of the ligand are all important for the distinct transcriptional responses to progesterone and cortisol of AR mutants, including the prostate cancer mutant L701H. PMID- 20007694 TI - The insulin-like growth factor-1 binding protein acid-labile subunit alters mesenchymal stromal cell fate. AB - Age-related osteoporosis is accompanied by an increase in marrow adiposity and a reduction in serum insulin-like growth factor-1 (IGF-1) and the binding proteins that stabilize IGF-1. To determine the relationship between these proteins and bone marrow adiposity, we evaluated the adipogenic potential of marrow-derived mesenchymal stromal cells (MSCs) from mice with decreased serum IGF-1 due to knockdown of IGF-1 production by the liver or knock-out of the binding proteins. We employed 10-16-week-old, liver-specific IGF-1-deficient, IGFBP-3 knock-out (BP3KO) and acid-labile subunit knock-out (ALSKO) mice. We found that expression of the late adipocyte differentiation marker peroxisome proliferator-activated receptor gamma was increased in marrow isolated from ALSKO mice. When induced with adipogenic media, MSC cultures from ALSKO mice revealed a significantly greater number of differentiated adipocytes compared with controls. MSCs from ALSKO mice also exhibited decreased alkaline-phosphatase positive colony size in cultures that were stimulated with osteoblast differentiation media. These osteoblast-like cells from ALSKO mice failed to induce osteoclastogenesis of control cells in co-culture assays, indicating that impairment of IGF-1 complex formation with ALS in bone marrow alters cell fate, leading to increased adipogenesis. PMID- 20007695 TI - Apoptotic death of hematopoietic tumor cells through potentiated and sustained adhesion to fibronectin via VLA-4. AB - It has been postulated that inactivated beta1-integrins are involved in the disordered growth of hematopoietic tumor cells. We recently found that TNIIIA2, a peptide derived from tenascin-C, strongly activates beta1-integrins through binding with syndecan-4. We show here that Ramos Burkitt's lymphoma cells can survive and grow in suspension but undergo apoptosis when kept adhering to fibronectin by stimulation with TNIIIA2. Other integrin activators, Mg(2+) and TS2/16 (an integrin-activating antibody), were also capable of inducing apoptosis. The inactivation of ERK1/2 and Akt and the subsequent activation of Bad were involved in the apoptosis. The results using other hematopoietic tumor cell lines expressing different levels of fibronectin receptors (VLA-4 and VLA-5) showed that potentiated and sustained adhesion to fibronectin via VLA-4 causally induces apoptosis also in various types of hematopoietic tumor cells in addition to Ramos cells. Because TNIIIA2 requires syndecan-4 as a membrane receptor for activation of beta1-integrins, it induced apoptosis preferentially in hematopoietic tumor cells, which expressed both VLA-4 and syndecan-4 as membrane receptors mediating the effects of fibronectin and TNIIIA2, respectively. Therefore, normal peripheral blood cells, such as neutrophils, monocytes, and lymphocytes, which poorly expressed syndecan-4, were almost insusceptible to TNIIIA2-induced apoptosis. The TNIIIA2-related matricryptic site of TN-C could contribute, once exposed, to preventing prolonged survival of hematopoietic malignant progenitors through potentiated and sustained activation of VLA-4. PMID- 20007696 TI - Catalytic role of the metal ion in the metallo-beta-lactamase GOB. AB - Metallo-beta-lactamases (MbetaLs) stand as one of the main mechanisms of bacterial resistance toward carbapenems. The rational design of an inhibitor for MbetaLs has been limited by an incomplete knowledge of their catalytic mechanism and by the structural diversity of their active sites. Here we show that the MbetaL GOB from Elizabethkingia meningoseptica is active as a monometallic enzyme by using different divalent transition metal ions as surrogates of the native Zn(II) ion. Of the metal derivatives in which Zn(II) is replaced, Co(II) and Cd(II) give rise to the most active enzymes and are shown to occupy the same binding site as the native ion. However, Zn(II) is the only metal ion capable of stabilizing an anionic intermediate that accumulates during nitrocefin hydrolysis, in which the C-N bond has already been cleaved. This finding demonstrates that the catalytic role of the metal ion in GOB is to stabilize the formation of this intermediate prior to nitrogen protonation. This role may be general to all MbetaLs, whereas nucleophile activation by a Zn(II) ion is not a conserved mechanistic feature. PMID- 20007697 TI - Opposing effects of two tissue transglutaminase protein isoforms in neuroblastoma cell differentiation. AB - We have demonstrated previously that the Myc oncoprotein blocks cancer cell differentiation by forming a novel transcriptional repressor complex with histone deacetylase and inhibiting gene transcription of tissue transglutaminase (TG2). Moreover, induction of TG2 gene transcription and transamidase activity is essential for the differentiating effects of retinoids in cancer cells. Here, we show that two structurally distinct TG2 protein isoforms, the full-length (TG2-L) and the short form (TG2-S), exert opposing effects on cell differentiation. Repression of TG2-L with small interfering RNA, which did not affect TG2-S expression, induced dramatic neuritic differentiation in neuroblastoma cells. In contrast, overexpression of TG2-S or a GTP-binding-deficient mutant of TG2-L (R580A), both of which lack the GTP-binding Arg-580 residue, induced neuroblastoma cell differentiation, which was blocked by an inhibitor of transamidase activity. Whereas N-Myc repressed and retinoid activated both TG2 isoforms, repression of TG2-L, but not simultaneous repression of TG2-L and TG2 S, enhanced neuroblastoma cell differentiation due to N-Myc small interfering RNA or retinoid. Moreover, suppression of vasoactive intestinal peptide (VIP) expression alone induced neuroblastoma cell differentiation, and VIP was up regulated by TG2-L, but not TG2-S. Taken together, our data indicate that TG2-L and TG2-S exert opposite effects on cell differentiation due to differences in GTP binding and modulation of VIP gene transcription. Our findings highlight the potential importance of repressing the GTP binding activity of TG2-L or activating the transamidase activity of TG2-L or TG2-S for the treatment of neuroblastoma, and possibly also other Myc-induced malignancies, and for enhancing retinoid anticancer effects. PMID- 20007698 TI - A network of hydrogen bonds on the surface of TLR2 controls ligand positioning and cell signaling. AB - TLR2 is a pattern recognition receptor that functions in association with TLR1 or TLR6 to mediate innate immune responses to a variety of conserved microbial products. In the present study, the ectodomain of TLR2 was extensively mutated, and the mutants were assessed for their ability to bind and to mediate cellular responses to triacylated lipopeptide Pam(3)CSK(4). This analysis provides evidence that the recently published crystal structure of the TLR2-TLR1 Pam(3)CSK(4) complex represents a functional signal-inducing complex. Furthermore, we report that extended H-bond networks on the surface of TLR2 are critical for signaling in response to Pam(3)CSK(4) and to other di- and tri acylated TLR2-TLR6 and TLR2-TLR1 ligands. Based on this finding, we suggest a dynamic model for TLR2-mediated recognition of these ligands in which TLR2 fluctuates between a conformation that is more suitable for binding of the fatty acyl moieties of the ligands and a conformation that favors, via a specific orientation of the ligand head group, formation of a signal-inducing ternary complex. PMID- 20007699 TI - BCR-ABL gene expression is required for its mutations in a novel KCL-22 cell culture model for acquired resistance of chronic myelogenous leukemia. AB - Acquired resistance through genetic mutations is a common phenomenon in several cancer therapies using molecularly targeted drugs, best exemplified by the BCR ABL inhibitor imatinib in treating chronic myelogenous leukemia (CML). Overcoming acquired resistance is a daunting therapeutic challenge, and little is known about how these mutations evolve. To facilitate understanding the resistance mechanisms, we developed a novel culture model for CML acquired resistance in which the CML cell line KCL-22, following initial response to imatinib, develops resistant T315I BCR-ABL mutation. We demonstrate that the emergence of BCR-ABL mutations do not require pre-existing BCR-ABL mutations derived from the original patient as the subclones of KCL-22 cells can form various BCR-ABL mutations upon imatinib treatment. BCR-ABL mutation rates vary from cell clone to clone and passages, in contrast to the relatively stable mutation rate of the hypoxanthine guanine phosphoribosyltransferase gene. Strikingly, development of BCR-ABL mutations depends on its gene expression because BCR-ABL knockdown completely blocks KCL-22 cell relapse on imatinib and acquisition of mutations. We further show that the endogenous BCR-ABL locus has significantly higher mutagenesis potential than the transduced randomly integrated BCR-ABL cDNA. Our study suggests important roles of BCR-ABL gene expression and its native chromosomal locus for acquisition of BCR-ABL mutations and provides a new tool for further studying resistance mechanisms. PMID- 20007700 TI - Cdc42p is activated during vacuole membrane fusion in a sterol-dependent subreaction of priming. AB - Cdc42p is a Rho GTPase that initiates signaling cascades at spatially defined intracellular sites for many cellular functions. We have previously shown that Cdc42p is localized to the yeast vacuole where it initiates actin polymerization during membrane fusion. Here we examine the activation cycle of Cdc42p during vacuole membrane fusion. Expression of either GTP- or GDP-locked Cdc42p mutants caused several morphological defects including enlarged cells and fragmented vacuoles. Stimulation of multiple rounds of fusion enhanced vacuole fragmentation, suggesting that cycles of Cdc42p activation, involving rounds of GTP binding and hydrolysis, are required to propagate Cdc42p signaling. We developed an assay to directly examine Cdc42p activation based on affinity to a probe derived from the p21-activated kinase effector, Ste20p. Cdc42p was rapidly activated during vacuole membrane fusion, which kinetically coincided with priming subreaction. During priming, Sec18p ATPase activity dissociates SNARE complexes and releases Sec17p, however, priming inhibitors such as Sec17p and Sec18p ligands did not block Cdc42p activation. Therefore, Cdc42p activation seems to be a parallel subreaction of priming, distinct from Sec18p activity. Specific mutants in the ergosterol synthesis pathway block both Sec17p release and Cdc42p activation. Taken together, our results define a novel sterol dependent subreaction of vacuole priming that activates cycles of Cdc42p activity to facilitate membrane fusion. PMID- 20007701 TI - Directed evolution reveals hidden properties of VMAT, a neurotransmitter transporter. AB - The vesicular neurotransmitter transporter VMAT2 is responsible for the transport of monoamines into synaptic and storage vesicles. VMAT2 is the target of many psychoactive drugs and is essential for proper neurotransmission and survival. Here we describe a new expression system in Saccharomyces cerevisiae that takes advantage of the polyspecificity of VMAT2. Expression of rVMAT2 confers resistance to acriflavine and to the parkinsonian toxin 1-methyl-4 phenylpyridinium (MPP(+)) by their removal into the yeast vacuole. This expression system allowed identification of a new substrate, acriflavine, and isolation of mutants with modified affinity to tetrabenazine (TBZ), a non competitive inhibitor of VMAT2 that is used in the treatment of various movement disorders including Tourette syndrome and Huntington chorea. Whereas one type of mutant obtained displayed decreased affinity to TBZ, a second type showed only a slight decrease in the affinity to TBZ, displayed a higher K(m) to the neurotransmitter serotonin, but conferred increased resistance to acriflavine and MPP(+). A protein where both types of mutations were combined (with only three amino acid replacements) lost most of the properties of the neurotransmitter transporter (TBZ-insensitive, no transport of neurotransmitter) but displayed enhanced resistance to the above toxicants. The work described here shows that in the case of rVMAT2, loss of traits acquired in evolution of function (such as serotonin transport and TBZ binding) bring about an improvement in older functions such as resistance to toxic compounds. A process that has taken millions of years of evolution can be reversed by three mutations. PMID- 20007703 TI - Niemann-Pick C1 functions independently of Niemann-Pick C2 in the initial stage of retrograde transport of membrane-impermeable lysosomal cargo. AB - The rare neurodegenerative disease Niemann-Pick Type C (NPC) results from mutations in either NPC1 or NPC2, which are membrane-bound and soluble lysosomal proteins, respectively. Previous studies have shown that mutations in either protein result in biochemically indistinguishable phenotypes, most notably the hyper-accumulation of cholesterol and other cargo in lysosomes. We comparatively evaluated the kinetics of [(3)H]dextran release from lysosomes of wild type, NPC1, NPC2, and NPC1/NPC2 pseudo-double mutant cells and found significant differences between all cell types examined. Specifically, NPC1 or NPC2 mutant fibroblasts treated with NPC1 or NPC2 siRNA (to create NPC1/NPC2 pseudo-double mutants) secreted dextran less efficiently than did either NPC1 or NPC2 single mutant cell lines, suggesting that the two proteins may work independently of one another in the egress of membrane-impermeable lysosomal cargo. To investigate the basis for these differences, we examined the role of NPC1 and NPC2 in the retrograde fusion of lysosomes with late endosomes to create so-called hybrid organelles, which is believed to be the initial step in the egress of cargo from lysosomes. We show here that cells with mutated NPC1 have significantly reduced rates of late endosome/lysosome fusion relative to wild type cells, whereas cells with mutations in NPC2 have rates that are similar to those observed in wild type cells. Instead of being involved in hybrid organelle formation, we show that NPC2 is required for efficient membrane fission events from nascent hybrid organelles, which is thought to be required for the reformation of lysosomes and the release of lysosomal cargo-containing membrane vesicles. Collectively, these results suggest that NPC1 and NPC2 can function independently of one another in the egress of certain membrane-impermeable lysosomal cargo. PMID- 20007702 TI - Antimitochondrial autoantibodies in pemphigus vulgaris: a missing link in disease pathophysiology. AB - A loss of epidermal cohesion in pemphigus vulgaris (PV) results from autoantibody action on keratinocytes (KCs) activating the signaling kinases and executioner caspases that damage KCs, causing their shrinkage, detachment from neighboring cells, and rounding up (apoptolysis). In this study, we found that PV antibody binding leads to activation of epidermal growth factor receptor kinase, Src, p38 MAPK, and JNK in KCs with time pattern variations from patient to patient. Both extrinsic and intrinsic apoptotic pathways were also activated. Although Fas ligand neutralizing antibody could inhibit the former pathway, the mechanism of activation of the latter remained unknown. PV antibodies increased cytochrome c release, suggesting damage to mitochondria. The immunoblotting experiments revealed penetration of PVIgG into the subcellular mitochondrial fraction. The antimitochondrial antibodies from different PV patients recognized distinct combinations of antigens with apparent molecular sizes of 25, 30, 35, 57, 60, and 100 kDa. Antimitochondrial antibodies were pathogenic because their absorption abolished the ability of PVIgG to cause keratinocyte detachment both in vitro and in vivo. The downstream signaling of antimitochondrial antibodies involved JNK and late p38 MAPK activation, whereas the signaling of anti-desmoglein 3 (Dsg3) antibody involved JNK and biphasic p38 MAPK activation. Using KCs grown from Dsg3(-/-) mice, we determined that Dsg3 did not serve as a surrogate antigen allowing antimitochondrial antibodies to enter KCs. The PVIgG-induced activation of epidermal growth factor receptor and Src was affected neither in Dsg3(-/-) KCs nor due to absorption of antimitochondrial antibodies. These results demonstrated that apoptolysis in PV is a complex process initiated by at least three classes of autoantibodies directed against desmosomal, mitochondrial, and other keratinocyte self-antigens. These autoantibodies synergize with the proapoptotic serum and tissue factors to trigger both extrinsic and intrinsic pathways of cell death and break the epidermal cohesion, leading to blisters. Further elucidation of the primary signaling events downstream of PV autoantigens will be crucial for the development of a more successful therapy for PV patients. PMID- 20007704 TI - A residue in loop 9 of the beta2-subunit stabilizes the closed state of the GABAA receptor. AB - In gamma-aminobutyric acid type A (GABA(A)) receptors, the structural elements that couple ligand binding to channel opening remain poorly defined. Here, site directed mutagenesis was used to determine if Loop 9 on the non-GABA binding site interface of the beta2-subunit may be involved in GABA(A) receptor activation. Specifically, residues Gly(170)-Gln(185) of the beta2-subunit were mutated to alanine, co-expressed with wild-type alpha1- and gamma2S-subunits in human embryonic kidney (HEK) 293 cells and assayed for their activation by GABA, the intravenous anesthetic propofol and the endogenous neurosteroid pregnanolone using whole cell macroscopic recordings. Three mutants, G170A, V175A, and G177A, produced 2.5-, 6.7-, and 5.6-fold increases in GABA EC(50) whereas one mutant, Q185A, produced a 5.2-fold decrease in GABA EC(50). None of the mutations affected the ability of propofol or pregnanolone to potentiate a submaximal GABA response, but the Q185A mutant exhibited 8.3- and 3.5-fold increases in the percent direct activation by propofol and pregnanolone, respectively. Mutant Q185A receptors also had an increased leak current that was sensitive to picrotoxin, indicating an increased gating efficiency. Further Q185E, Q185L, and Q185W substitutions revealed a strong correlation between the hydropathy of the amino acid at this position and the GABA EC(50). Taken together, these results indicate that beta2 Loop 9 is involved in receptor activation by GABA, propofol, and pregnanolone and that beta2(Q185) participates in hydrophilic interactions that are important for stabilizing the closed state of the GABA(A) receptor. PMID- 20007705 TI - Evidences for a leaky scanning mechanism for the synthesis of the shorter M23 protein isoform of aquaporin-4: implication in orthogonal array formation and neuromyelitis optica antibody interaction. AB - Aquaporin-4 (AQP4) exists as two major isoforms that differ in the length of the N terminus, the shorter AQP4-M23 and the longer AQP4-M1. Both isoforms form tetramers, which can further aggregate in the plasma membrane to form typical orthogonal arrays of particles (OAPs) whose dimension depends on the ratio of the M1 and M23. In this study, we tested the hypothesis that the M23 isoform can be produced directly by the M1 mRNA. In cells transiently transfected with AQP4-M1 coding sequence we observed besides AQP4-M1 the additional presence of the AQP4 M23 isoform associated with the formation of typical OAPs observable by two dimensional blue native/SDS-PAGE and total internal reflection microscopy. The mutation of the second in-frame methionine M23 in AQP4-M1 (AQP4-M1(M23I)) prevented the expression of the M23 isoform and the formation of OAPs. We propose "leaky scanning" as a translational mechanism for the expression of AQP4-M23 protein isoform and that the formation of OAPs may occur even in the absence of AQP4-M23 mRNA. This mechanism can have important pathophysiological implications for the cell regulation of the M1/M23 ratio and thus OAP size. In this study we also provide evidence that AQP4-M1 is mobile in the plasma membrane, that it is inserted and not excluded into immobile OAPs, and that it is an important determinant of OAP structure and size. PMID- 20007706 TI - FGF2-activated ERK mitogen-activated protein kinase enhances Runx2 acetylation and stabilization. AB - Runx2 is a key transcription factor regulating osteoblast differentiation and skeletal morphogenesis, and FGF2 is one of the most important regulators of skeletal development. The importance of the ERK mitogen-activated protein (MAP) kinase pathway in cranial suture development was demonstrated by the findings that the inhibition of FGF/FGF receptor (FGFR) signaling by a MEK blocker prevents the premature suture closure caused by an Fgfr2 mutation in mice. We previously demonstrated that ERK activation does not affect Runx2 gene expression but that it stimulates Runx2 transcriptional activity. However, the molecular mechanism underlying Runx2 activation by FGF/FGFR or ERK was still unclear. In this study, we found that FGF2 treatment increased the protein level of exogenously overexpressed Runx2 and that this increase is reversed by ERK inhibitors. In contrast, overexpression of constitutively active MEK strongly increased the Runx2 protein level, which paralleled an increase in Runx2 acetylation. As Runx2 protein phosphorylation mediated by ERK directly correlates with Runx2 protein stabilization, acetylation, and ubiquitination, we undertook to identify the ERK-dependent phosphorylation sites in Runx2. Analysis of two C terminal Runx2 deletion constructs showed that the middle third of the protein is responsible for ERK-induced stabilization and activation. An in silico analysis of highly conserved ERK targets indicated that there are three relevant serine residues in this domain. Site-directed mutagenesis implicated Ser-301 in for ERK mediated Runx2 stabilization and acetylation. In conclusion, the FGF2-induced ERK MAP kinase strongly increased the Runx2 protein level through an increase in acetylation and a decrease in ubiquitination, and these processes require the phosphorylation of Runx2 Ser-301 residue. PMID- 20007707 TI - The protein-protein interface evolution acts in a similar way to antibody affinity maturation. AB - Understanding the evolutionary mechanism that acts at the interfaces of protein protein complexes is a fundamental issue with high interest for delineating the macromolecular complexes and networks responsible for regulation and complexity in biological systems. To investigate whether the evolution of protein-protein interface acts in a similar way as antibody affinity maturation, we incorporated evolutionary information derived from antibody affinity maturation with common simulation techniques to evaluate prediction success rates of the computational method in affinity improvement in four different systems: antibody-receptor, antibody-peptide, receptor-membrane ligand, and receptor-soluble ligand. It was interesting to find that the same evolutionary information could improve the prediction success rates in all the four protein-protein complexes with an exceptional high accuracy (>57%). One of the most striking findings in our present study is that not only in the antibody-combining site but in other protein-protein interfaces almost all of the affinity-enhancing mutations are located at the germline hotspot sequences (RGYW or WA), indicating that DNA hot spot mechanisms may be widely used in the evolution of protein-protein interfaces. Our data suggest that the evolution of distinct protein-protein interfaces may use the same basic strategy under selection pressure to maintain interactions. Additionally, our data indicate that classical simulation techniques incorporating the evolutionary information derived from in vivo antibody affinity maturation can be utilized as a powerful tool to improve the binding affinity of protein-protein complex with a high accuracy. PMID- 20007708 TI - Adipocyte-mononuclear cell interaction, Toll-like receptor 4 activation, and high glucose synergistically up-regulate osteopontin expression via an interleukin 6 mediated mechanism. AB - Although it has been reported that osteopontin, a matrix glycoprotein and proinflammatory cytokine, mediates obesity-induced adipose tissue macrophage infiltration and insulin resistance, it remains unclear how osteopontin is up regulated in adipose tissue in obese humans and animals. In this study, we incubated U937 mononuclear cells with adipocytes in a transwell system and studied how cell interaction regulated osteopontin expression. Results showed that coculture of U937 cells with adipocytes led to a marked increase in osteopontin production when compared with that released by independent cultures of U937 cells. Moreover, lipopolysaccharide or palmitic acid-induced TLR4 activation and high glucose further augmented the coculture-stimulated osteopontin secretion. Similar observations were made in the coculture of human primary monocytes and adipocytes. Real time PCR studies showed that coculture of U937 cells and adipocytes increased osteopontin mRNA in U937 cells, but not adipocytes, suggesting that adipocyte-derived soluble factor may stimulate osteopontin expression by U937 cells. In our studies to explore the underlying mechanism, we found that the neutralizing antibodies against interleukin (IL)-6 or IL-6 small interfering RNA transfection in adipocytes effectively inhibited coculture-stimulated osteopontin expression, suggesting that IL-6 released by adipocytes plays an essential role in the coculture-stimulated osteopontin expression by U937 cells. In conclusion, this study has demonstrated that cell interaction, TLR4 activation, and high glucose up-regulate osteopontin expression, and adipocyte-derived IL-6 played a major role in the up-regulation. PMID- 20007709 TI - SOCS-6 negatively regulates T cell activation through targeting p56lck to proteasomal degradation. AB - The T cell-specific tyrosine kinase, p56(lck), plays crucial roles in T cell receptor (TCR)-mediated T cell activation. Here, we report that SOCS-6 (suppressor of cytokine signaling-6) is a negative regulator of p56(lck). SOCS-6 was identified as a protein binding to the kinase domain of p56(lck) through yeast two-hybrid screening. SOCS-6 bound specifically to p56(lck) (F505), which mimics the active form of p56(lck), but not to wild type p56(lck). In Jurkat T cells, SOCS-6 binding to p56(lck) was detected 1-2 h after TCR stimulation. Confocal microscopy showed that upon APC-T cell conjugation, SOCS-6 was recruited to the immunological synapse and colocalized with the active form of p56(lck). SOCS-6 promoted p56(lck) ubiquitination and its subsequent targeting to the proteasome. Moreover, SOCS-6 overexpression led to repression of TCR-dependent interleukin-2 promoter activity. These results establish that SOCS-6 acts as a negative regulator of T cell activation by promoting ubiquitin-dependent proteolysis. PMID- 20007710 TI - The mitochondrial pathway of anesthetic isoflurane-induced apoptosis. AB - The common inhalation anesthetic isoflurane has been shown to induce apoptosis, which then leads to accumulation of beta-amyloid protein, the hallmark feature of Alzheimer disease neuropathogenesis. The underlying molecular mechanism of the isoflurane-induced apoptosis is largely unknown. We, therefore, set out to assess whether isoflurane can induce apoptosis by regulating Bcl-2 family proteins, enhancing reactive oxygen species (ROS) accumulation, and activating the mitochondrial pathway of apoptosis. We performed these studies in cultured cells, primary neurons, and mice. Here we show for the first time that treatment with 2% isoflurane for 6 h can increase pro-apoptotic factor Bax levels, decrease anti apoptotic factor Bcl-2 levels, increase ROS accumulation, facilitate cytochrome c release from the mitochondria to the cytosol, induce activation of caspase-9 and caspase-3, and finally cause apoptosis as compared with the control condition. We have further found that isoflurane can increase the mRNA levels of Bax and reduce the mRNA levels of Bcl-2. The isoflurane-induced ROS accumulation can be attenuated by the intracellular calcium chelator BAPTA. Finally, the anesthetic desflurane does not induce activation of mitochondrial pathway of apoptosis. These results suggest that isoflurane may induce apoptosis through Bcl-2 family proteins- and ROS-associated mitochondrial pathway of apoptosis. These findings, which have identified at least partially the molecular mechanism by which isoflurane induces apoptosis, will promote more studies aimed at studying the potential neurotoxic effects of anesthetics. PMID- 20007711 TI - The intracellular loop of Orai1 plays a central role in fast inactivation of Ca2+ release-activated Ca2+ channels. AB - Store-operated Ca(2+) entry (SOCE) due to activation of Ca(2+) release-activated Ca(2+) (CRAC) channels leads to sustained elevation of cytoplasmic Ca(2+) and activation of lymphocytes. CRAC channels consisting of four pore-forming Orai1 subunits are activated by STIM1, an endoplasmic reticulum Ca(2+) sensor that senses intracellular store depletion and migrates to plasma membrane proximal regions to mediate SOCE. One of the fundamental properties of CRAC channels is their Ca(2+)-dependent fast inactivation. To identify the domains of Orai1 involved in fast inactivation, we have mutated residues in the Orai1 intracellular loop linking transmembrane segment II to III. Mutation of four residues, V(151)SNV(154), at the center of the loop (MutA) abrogated fast inactivation, leading to increased SOCE as well as higher CRAC currents. Point mutation analysis identified five key amino acids, N(153)VHNL(157), that increased SOCE in Orai1 null murine embryonic fibroblasts. Expression or direct application of a peptide comprising the entire intracellular loop or the sequence N(153)VHNL(157) blocked CRAC currents from both wild type (WT) and MutA Orai1. A peptide incorporating the MutA mutations had no blocking effect. Concatenated Orai1 constructs with four MutA monomers exhibited high CRAC currents lacking fast inactivation. Reintroduction of a single WT monomer (MutA-MutA-MutA-WT) was sufficient to fully restore fast inactivation, suggesting that only a single intracellular loop can block the channel. These data suggest that the intracellular loop of Orai1 acts as an inactivation particle, which is stabilized in the ion permeation pathway by the N(153)VHNL(157) residues. These results along with recent reports support a model in which the N terminus and the selectivity filter of Orai1 as well as STIM1 act in concert to regulate the movement of the intracellular loop and evoke fast inactivation. PMID- 20007712 TI - Differential regulation of phospholipase C-beta2 activity and membrane interaction by Galphaq, Gbeta1gamma2, and Rac2. AB - We combined fluorescence recovery after photobleaching (FRAP) beam-size analysis with biochemical assays to investigate the mechanisms of membrane recruitment and activation of phospholipase C-beta(2) (PLCbeta(2)) by G protein alpha(q) and betagamma dimers. We show that activation by alpha(q) and betagamma differ from activation by Rac2 and from each other. Stimulation by alpha(q) enhanced the plasma membrane association of PLCbeta(2), but not of PLCbeta(2)Delta, which lacks the alpha(q)-interacting region. Although alpha(q) resembled Rac2 in increasing the contribution of exchange to the FRAP of PLCbeta(2) and in enhancing its membrane association, the latter effect was weaker than with Rac2. Moreover, the membrane recruitment of PLCbeta(2) by alpha(q) occurred by enhancing PLCbeta(2) association with fast-diffusing (lipid-like) membrane components, whereas stimulation by Rac2 led to interactions with slow diffusing membrane sites. On the other hand, activation by betagamma shifted the FRAP of PLCbeta(2) and PLCbeta(2)Delta to pure lateral diffusion 3- to 5-fold faster than lipids, suggesting surfing-like diffusion along the membrane. We propose that these different modes of PLCbeta(2) membrane recruitment may accommodate contrasting functional needs to hydrolyze phosphatidylinositol 4,5-bisphosphate (PtdInsP(2)) in localized versus dispersed populations. PLCbeta(2) activation by Rac2, which leads to slow lateral diffusion and much faster exchange, recruits PLCbeta(2) to act locally on PtdInsP(2) at specific domains. Activation by alpha(q) leads to lipid-like diffusion of PLCbeta(2) accompanied by exchange, enabling the sampling of larger, yet limited, areas prior to dissociation. Finally, activation by betagamma recruits PLCbeta(2) to the membrane by transient interactions, leading to fast "surfing" diffusion along the membrane, sampling large regions for dispersed PtdInsP(2) populations. PMID- 20007713 TI - A structural element within the HUWE1 HECT domain modulates self-ubiquitination and substrate ubiquitination activities. AB - E3 ubiquitin ligases catalyze the final step of ubiquitin conjugation and regulate numerous cellular processes. The HECT class of E3 ubiquitin (Ub) ligases directly transfers Ub from bound E2 enzyme to a myriad of substrates. The catalytic domain of HECT Ub ligases has a bilobal architecture that separates the E2 binding region and catalytic site. An important question regarding HECT domain function is the control of ligase activity and specificity. Here we present a functional analysis of the HECT domain of the E3 ligase HUWE1 based on crystal structures and show that a single N-terminal helix significantly stabilizes the HECT domain. We observe that this element modulates HECT domain activity, as measured by self-ubiquitination induced in the absence of this helix, as distinct from its effects on Ub conjugation of substrate Mcl-1. Such subtle changes to the protein may be at the heart of the vast spectrum of substrate specificities displayed by HECT domain E3 ligases. PMID- 20007714 TI - ATPase domain and interdomain linker play a key role in aggregation of mitochondrial Hsp70 chaperone Ssc1. AB - The co-chaperone Hep1 is required to prevent the aggregation of mitochondrial Hsp70 proteins. We have analyzed the interaction of Hep1 with mitochondrial Hsp70 (Ssc1) and the determinants in Ssc1 that make it prone to aggregation. The ATPase and peptide binding domain (PBD) of Hsp70 proteins are connected by a linker segment that mediates interdomain communication between the domains. We show here that the minimal Hep1 binding entity of Ssc1 consists of the ATPase domain and the interdomain linker. In the absence of Hep1, the ATPase domain with the interdomain linker had the tendency to aggregate, in contrast to the ATPase domain with the mutated linker segment or without linker, and in contrast to the PBD. The closest homolog of Ssc1, bacterial DnaK, and a Ssc1 chimera, in which a segment of the ATPase domain of Ssc1 was replaced by the corresponding segment from DnaK, did not aggregate in Delta hep1 mitochondria. The propensity to aggregate appears to be a specific property of the mitochondrial Hsp70 proteins. The ATPase domain in combination with the interdomain linker is crucial for aggregation of Ssc1. In conclusion, our results suggest that interdomain communication makes Ssc1 prone to aggregation. Hep1 counteracts aggregation by binding to this aggregation-prone conformer. PMID- 20007716 TI - Structural basis for translation factor recruitment to the eukaryotic/archaeal ribosomes. AB - The archaeal ribosomal stalk complex has been shown to have an apparently conserved functional structure with eukaryotic pentameric stalk complex; it provides access to eukaryotic elongation factors at levels comparable to that of the eukaryotic stalk. The crystal structure of the archaeal heptameric (P0(P1)(2)(P1)(2)(P1)(2)) stalk complex shows that the rRNA anchor protein P0 consists of an N-terminal rRNA-anchoring domain followed by three separated spine helices on which three P1 dimers bind. Based on the structure, we have generated P0 mutants depleted of any binding site(s) for P1 dimer(s). Factor-dependent GTPase assay of such mutants suggested that the first P1 dimer has higher activity than the others. Furthermore, we constructed a model of the archaeal 50 S with stalk complex by superposing the rRNA-anchoring domain of P0 on the archaeal 50 S. This model indicates that the C termini of P1 dimers where translation factors bind are all localized to the region between the stalk base of the 50 S and P0 spine helices. Together with the mutational experiments we infer that the functional significance of multiple copies of P1 is in creating a factor pool within a limited space near the stalk base of the ribosome. PMID- 20007715 TI - Amino acid residues critical for endoplasmic reticulum export and trafficking of platelet-activating factor receptor. AB - Several residues are conserved in the transmembrane domains (TMs) of G-protein coupled receptors. Here we demonstrate that a conserved proline, Pro(247), in TM6 of platelet-activating factor receptor (PAFR) is required for endoplasmic reticulum (ER) export and trafficking after agonist-induced internalization. Alanine-substituted mutants of the conserved residues of PAFRs, including P247A, were retained in the ER. Because a PAFR antagonist, Y-24180, acted as a pharmacological chaperone to rescue ER retention, this retention is due to misfolding of PAFR. Methylcarbamyl (mc)-PAF, a PAFR agonist, did not increase the cell surface expression of P247A, even though another ER-retained mutant, D63A, was effectively trafficked. Signaling and accumulation of the receptors in the early endosomes were observed in the mc-PAF-treated P247A-expressing cells, suggesting that P247A was trafficked to the cell surface by mc-PAF, and thereafter disappeared from the surface due to aberrant trafficking, e.g. enhanced internalization, deficiency in recycling, and/or accelerated degradation. The aberrant trafficking was confirmed with a sortase-A-mediated method for labeling cell surface proteins. These results demonstrate that the conserved proline in TM6 is crucial for intracellular trafficking of PAFR. PMID- 20007717 TI - Molecular characterization of the interaction of staphylococcal microbial surface components recognizing adhesive matrix molecules (MSCRAMM) ClfA and Fbl with fibrinogen. AB - The ligand-binding domain of Fbl (the fibrinogen binding protein from Staphylococcus lugdunensis) shares 60% sequence identity with ClfA (clumping factor A) of Staphylococcus aureus. Recombinant Fbl corresponding to the minimum fibrinogen-binding region (subdomains N2N3) was compared with ClfA for binding to fibrinogen. Fbl and ClfA had very similar affinities for fibrinogen by surface plasmon resonance. The binding site for Fbl in fibrinogen was localized to the extreme C terminus of the fibrinogen gamma-chain at the same site recognized by ClfA. Isothermal titration calorimetry showed that Fbl and ClfA had very similar affinities for a peptide mimicking the C-terminal segment of the fibrinogen gamma chain. The peptide also inhibited binding of Fbl and ClfA to fibrinogen. A series of substituted gamma-chain variant peptides behaved very similarly when used to inhibit ClfA and Fbl binding to immobilized fibrinogen. Both ClfA and Fbl bound to bovine fibrinogen with a lower affinity compared with human fibrinogen and did not bind detectably to ovine fibrinogen. The structure of the N2N3 subdomains of Fbl in complex with the fibrinogen gamma-chain peptide was modeled based on the crystal structure of the N2N3 subdomains of the ClfA-gamma-chain peptide complex. Residues in the putative binding trench likely to be involved in fibrinogen binding were identified. Fbl variant proteins with alanine substitutions in key residues had reduced affinities for fibrinogen. Thus Fbl and ClfA bind the same site in fibrinogen by similar mechanisms. PMID- 20007718 TI - Conditional Niemann-Pick C mice demonstrate cell autonomous Purkinje cell neurodegeneration. AB - Pathways regulating neuronal vulnerability are poorly understood, yet are central to identifying therapeutic targets for degenerative neurological diseases. Here, we characterize mechanisms underlying neurodegeneration in Niemann-Pick type C (NPC) disease, a lysosomal storage disorder characterized by impaired cholesterol trafficking. To date, the relative contributions of neuronal and glial defects to neuron loss are poorly defined. Using gene targeting, we generate Npc1 conditional null mutant mice. Deletion of Npc1 in mature cerebellar Purkinje cells leads to an age-dependent impairment in motor tasks, including rotarod and balance beam performance. Surprisingly, these mice did not show the early death or weight loss that are characteristic of global Npc1 null mice, suggesting that Purkinje cell degeneration does not underlie these phenotypes. Histological examination revealed the progressive loss of Purkinje cells in an anterior-to posterior gradient. This cell autonomous neurodegeneration occurs in a spatiotemporal pattern similar to that of global knockout mice. A subpopulation of Purkinje cells in the posterior cerebellum exhibits marked resistance to cell death despite Npc1 deletion. To explore this selective response, we investigated the electrophysiological properties of vulnerable and susceptible Purkinje cell subpopulations. Unexpectedly, Purkinje cells in both subpopulations displayed no electrophysiological abnormalities prior to degeneration. Our data establish that Npc1 deficiency leads to cell autonomous, selective neurodegeneration and suggest that the ataxic symptoms of NPC disease arise from Purkinje cell death rather than cellular dysfunction. PMID- 20007719 TI - Tuberculous endocarditis: valvular and right atrial involvement. AB - Valvular endocarditis due to Mycobacterium tuberculosis is a rare clinical entity. It is usually manifest in the context of disseminated tuberculosis in immunocompromised patients. This report describes a unique case of a 30-year-old immunocompetent man with an incidental finding of tuberculous valvular endocarditis. The patient had a large mass on the anterior mitral leaflet and severe mitral regurgitation. He underwent mitral valve replacement and Mycobacterium tuberculosis was cultured from the valve vegetation and the right atrial masses. Post-operative recovery has been uneventful without relapse for 24 months. PMID- 20007720 TI - Revisiting imaging features and the embryologic basis of third and fourth branchial anomalies. AB - BACKGROUND AND PURPOSE: There is wide discrepancy between common clinical and radiologic presentations of branchial sinuses arising from the pyriform fossa and the theoretic course of third and fourth branchial arch anomalies. The purpose of this study was to revisit the clinical presentations and imaging features of such anomalies in children. MATERIALS AND METHODS: A retrospective review of institutional and diagnostic imaging data bases from 1998 to 2008 for reported cases of third and fourth branchial cleft anomalies was conducted. Clinical presentation, pharyngoscopy results, and imaging features in all the patients were evaluated. Surgical and histopathology correlation in patients who underwent excision of the tract was also obtained. RESULTS: Twenty reported cases described as third or fourth branchial apparatus anomalies were identified. There were 12 females and 8 males with a mean age of 84.6 months. The most common presentation was an inflammatory neck mass (18/20, 90%) almost always involving the thyroid gland. Most lesions were on the left side (16/20, 80%). Pharyngoscopy showed a sinus opening at the piriform fossa in 18/20 (90%) cases. None of the cases followed the classic theoretic pathway of third and fourth arch remnants. Histopathology showed tracts lined with pseudostratified squamous epithelium or ciliated columnar epithelium often associated with inflammatory changes in 17 surgically resected cases. CONCLUSIONS: Branchial sinuses arising from the pyriform fossa often present with an inflammatory neck mass involving the thyroid lobe, most often on the left side. Imaging and surgical findings suggest that they arise from the embryonal thymopharyngeal duct of the third branchial pouch, because they do not follow the hypothetic course of third or fourth arch fistulas. PMID- 20007721 TI - CT features of lobular capillary hemangioma of the nasal cavity. AB - BACKGROUND AND PURPOSE: Lobular capillary hemangioma is a benign capillary proliferation of unknown etiology. To our knowledge, no comprehensive review of imaging findings of LCHNC has been presented. Thus, we investigated characteristic CT features of LCHNC. MATERIALS AND METHODS: This retrospective study included 6 patients (2 men and 4 women; age range, 30-65 years; mean age, 49.2 years) with histologically proved LCHNC. We evaluated the size, site of origin, attenuation on NECT, degree and pattern of enhancement, and bony changes. RESULTS: The LCHNC lesion was 13.0-45.0 mm (average, 25.0 mm) in diameter. These lesions arose from the inferior turbinate in 5 (83.3%) patients and the anterior nasal septum in 1 (16.7%). Compared with the masticator muscles, the LCHNC lesion was hypoattenuating in 2 (33.3%) and isoattenuating on NECT in 4 (66.7%) patients. In 5 (83.3%) patients, the LCHNC lesion consisted of 2 distinct areas on CECT: a lobular intensely enhancing mass and an iso- or hypoattenuating cap of variable thickness around the intensely enhancing mass. Bony changes included erosion in 3 (50.0%) and displacement in 2 (33.3%) patients. CONCLUSIONS: CT features of LCHNC consist of an intensely enhancing mass and an iso- or hypoattenuating cap on CECT. The inferior turbinate seems to be a common site of origin, and bony changes are not uncommon features of LCHNC. CT is useful not only in identifying the site of origin and assessing the extent but also in suggesting the nature of LCHNC. PMID- 20007722 TI - Transient hemiglossal denervation during acute internal capsule infarct in the setting of dysarthria-clumsy hand syndrome. AB - A case of MR imaging-documented transient unilateral tongue denervation presenting during acute internal capsule infarction is described. Understanding the corticolingual pathway innervation of the hypoglossal nucleus is essential for explaining these findings. Awareness of the findings in this case will facilitate appropriate diagnosis, provide neuroanatomic explanation, and prevent misdiagnosis. PMID- 20007723 TI - Utility of the k-means clustering algorithm in differentiating apparent diffusion coefficient values of benign and malignant neck pathologies. AB - BACKGROUND AND PURPOSE: Does the K-means algorithm do a better job of differentiating benign and malignant neck pathologies compared to only mean ADC? The objective of our study was to analyze the differences between ADC partitions to evaluate whether the K-means technique can be of additional benefit to whole lesion mean ADC alone in distinguishing benign and malignant neck pathologies. MATERIAL AND METHODS: MR imaging studies of 10 benign and 10 malignant proved neck pathologies were postprocessed on a PC by using in-house software developed in Matlab. Two neuroradiologists manually contoured the lesions, with the ADC values within each lesion clustered into 2 (low, ADC-ADC(L); high, ADC-ADC(H)) and 3 partitions (ADC(L); intermediate, ADC-ADC(I); ADC(H)) by using the K-means clustering algorithm. An unpaired 2-tailed Student t test was performed for all metrics to determine statistical differences in the means of the benign and malignant pathologies. RESULTS: A statistically significant difference between the mean ADC(L) clusters in benign and malignant pathologies was seen in the 3 cluster models of both readers (P = .03 and .022, respectively) and the 2-cluster model of reader 2 (P = .04), with the other metrics (ADC(H), ADC(I); whole-lesion mean ADC) not revealing any significant differences. ROC curves demonstrated the quantitative differences in mean ADC(H) and ADC(L) in both the 2- and 3-cluster models to be predictive of malignancy (2 clusters: P = .008, area under curve = 0.850; 3 clusters: P = .01, area under curve = 0.825). CONCLUSIONS: The K-means clustering algorithm that generates partitions of large datasets may provide a better characterization of neck pathologies and may be of additional benefit in distinguishing benign and malignant neck pathologies compared with whole-lesion mean ADC alone. PMID- 20007724 TI - High-b-value diffusion MR imaging and basal nuclei apparent diffusion coefficient measurements in variant and sporadic Creutzfeldt-Jakob disease. AB - BACKGROUND AND PURPOSE: DWI using a standard b-value of 1000 s/mm(2) has emerged as the most sensitive sequence for the diagnosis of CJD. The purpose of this study was to investigate whether DWI at a high b-value (b = 3000 s/mm(2)) and ADC measurements in the basal nuclei improve the diagnosis of vCJD and sCJD compared with visual assessment of DWI at a standard b-value (b = 1000 s/mm(2)). MATERIALS AND METHODS: Eight patients with vCJD, 9 patients with sCJD, and 5 healthy volunteers underwent DWI at b = 1000 s/mm(2), and 5 vCJD patients, 4 sCJD patients, and 1 growth hormone-related CJD patient underwent DWI at b = 3000 s/mm(2). Two consultant neuroradiologists performed a visual comparison of the b = 1000 and b = 3000 images. Mean MR SI and ADC values were determined for C, P, and DM thalamus ROIs bilaterally at each b-value. SI ratios for each ROI relative to white matter were calculated. RESULTS: In 9 out of 10 patients, the higher b value images were more sensitive to SI change, particularly in cortex and thalamus, with higher SI ratios at b = 3000 in the DM thalamus. For sCJD at b = 1000, we found significantly lower ADC values in the C and P compared with controls (mean C ADC = 587.3 +/- 84.7 mm(2)/s in sCJD patients versus 722.7 +/- 16.6 mm(2)/s in controls; P = .007), and at b = 3000, the differences were more pronounced. In comparison, in vCJD at b = 1000, ADC values were elevated in the Pu (mean Pu ADC = 837.6 +/- 33.0 mm/s(2) in vCJD patients versus 748.0 +/- 17.3 mm/s(2) in controls; P < .001) but failed to reach significance at b = 3000. CONCLUSIONS: Our results demonstrate that b = 3000 DWI, being more sensitive to slowly diffusing tissue water, is more sensitive to pathology in sCJD than is conventional DWI. High-b-value DWI increases confidence in the radiologic diagnosis of human prion disease. PMID- 20007725 TI - Characterizing brain tumor research: the role of the National Institutes of Health. AB - The contribution of radiology to brain tumor research is unknown. We sought to determine how the proportion of neuro-oncologic publications generated by radiology departments has changed and if there is an association with NIH funding levels. Therefore we searched The National Library of Medicine's PubMed database for all articles published on brain neoplasms from 1996 to 2007. Country and department of origin and NIH grant support were noted for each article. Approximately 10% of brain tumor publications originated from radiology departments, ranking third among medical specialties. NIH funding for this research grew from less than 20% in 1996 to more than 50% in 2007. Overall NIH funding levels rose approximately 2.5 fold during this time. The U.S. was the dominant producer of brain tumor publications, and the gap between the U.S. and the rest of the world grew over the study period. Thus a substantial proportion of brain tumor publications originate from radiology departments, and the percentage of this research that is funded by the NIH has grown significantly during a period of increasing NIH budgets. PMID- 20007726 TI - Insights into adult postlesional language cortical plasticity provided by cerebral blood oxygen level-dependent functional MR imaging. AB - BOLD fMRI has provided new insights into postlesional brain language plasticity by providing a noninvasive in vivo approach to evaluate longitudinal changes in brain cortical activation during performance of language tasks. Specifically, BOLD fMRI has provided the opportunity to investigate not only changes in eloquent language cortex resulting from different types of brain pathology such as brain tumors, stroke, and epilepsy but also changes in eloquent language cortex occurring as a result of actual surgical resection of diseased but, nevertheless, partially functional tissue. In addition to reviewing the literature relating to stroke and epilepsy-related language plasticity as well as the more intriguing phenomenon of postsurgical plasticity in the setting of brain tumors, 2 unusual cases illustrating this latter manifestation of language plasticity are briefly described in this review article. PMID- 20007727 TI - A novel association between RASA1 mutations and spinal arteriovenous anomalies. AB - BACKGROUND AND PURPOSE: CM-AVM is a recently recognized autosomal dominant disorder associated with mutations in RASA1. Arteriovenous lesions have been reported in the brain, limbs, and the face in 18.5% of patients. We report a novel association between RASA1 mutations and spinal arteriovenous anomalies. MATERIALS AND METHODS: In a collaborative study, 5 index patients (2 females, 3 males) with spinal AVMs or AVFs and cutaneous multifocal capillary lesions were investigated for the RASA1 gene mutation. RESULTS: All 5 patients were found to have RASA1 mutation (2 de novo, 3 familial), and all had multifocal capillary malformations at birth. Neurologic deficits developed at ages ranging from infancy to early adulthood. All spinal anomalies (2 AVMs at the conus, 1 AVM at the lumbosacral junction, and 1 cervical and 1 cervicothoracic AVF) were complex, extensive, and fast-flow lesions. All patients required treatment based on the clinical and/or radiologic appearance of the lesions. CONCLUSIONS: To our knowledge, an association of RASA1 mutation and spinal AVM/AVF has not been described. MR imaging screening of patients with characteristic CMs and neurologic symptoms presenting at a young age may be useful in detecting the presence of fast-flow intracranial or intraspinal arteriovenous anomalies before potentially significant neurologic insult has occurred. PMID- 20007729 TI - Bioinformatical approaches to characterize intrinsically disordered/unstructured proteins. AB - Intrinsically disordered/unstructured proteins exist without a stable three dimensional (3D) structure as highly flexible conformational ensembles. The available genome sequences revealed that these proteins are surprisingly common and their frequency reaches high proportions in eukaryotes. Due to their vital role in various biological processes including signaling and regulation and their involvement in various diseases, disordered proteins and protein segments are the focus of many biochemical, molecular biological, pathological and pharmaceutical studies. These proteins are difficult to study experimentally because of the lack of unique structure in the isolated form. Their amino acid sequence, however, is available, and can be used for their identification and characterization by bioinformatic tools, analogously to globular proteins. In this review, we first present a small survey of current methods to identify disordered proteins or protein segments, focusing on those that are publicly available as web servers. In more detail we also discuss approaches that predict disordered regions and specific regions involved in protein binding by modeling the physical background of protein disorder. In our review we argue that the heterogeneity of disordered segments needs to be taken into account for a better understanding of protein disorder. PMID- 20007728 TI - Advances in translational bioinformatics: computational approaches for the hunting of disease genes. AB - Over a 100 years ago, William Bateson provided, through his observations of the transmission of alkaptonuria in first cousin offspring, evidence of the application of Mendelian genetics to certain human traits and diseases. His work was corroborated by Archibald Garrod (Archibald AE. The incidence of alkaptonuria: a study in chemical individuality. Lancert 1902;ii:1616-20) and William Farabee (Farabee WC. Inheritance of digital malformations in man. In: Papers of the Peabody Museum of American Archaeology and Ethnology. Cambridge, Mass: Harvard University, 1905; 65-78), who recorded the familial tendencies of inheritance of malformations of human hands and feet. These were the pioneers of the hunt for disease genes that would continue through the century and result in the discovery of hundreds of genes that can be associated with different diseases. Despite many ground-breaking discoveries during the last century, we are far from having a complete understanding of the intricate network of molecular processes involved in diseases, and we are still searching for the cures for most complex diseases. In the last few years, new genome sequencing and other high-throughput experimental techniques have generated vast amounts of molecular and clinical data that contain crucial information with the potential of leading to the next major biomedical discoveries. The need to mine, visualize and integrate these data has motivated the development of several informatics approaches that can broadly be grouped in the research area of 'translational bioinformatics'. This review highlights the latest advances in the field of translational bioinformatics, focusing on the advances of computational techniques to search for and classify disease genes. PMID- 20007730 TI - Reduction in erythromycin resistance in invasive pneumococci from young children in England and Wales. PMID- 20007732 TI - Sensory testing of distal sural and posterior tibial nerves provides early prediction of surgical anesthesia after single-injection infragluteal-parabiceps sciatic nerve block. AB - BACKGROUND: Surgical anesthesia for reconstructive ankle surgery requires sensory and motor block of all the terminal nerve distributions of the sciatic nerve. In this prospective observational study, we investigated the value of sensory and motor testing of the foot, after local anesthetic injection, for predicting complete sciatic nerve blockade and the duration of testing required for identifying incomplete anesthesia. METHODS: Sciatic nerve blocks (n = 180) using the infragluteal-parabiceps approach were performed in patients undergoing reconstructive ankle surgery. Levobupivacaine 0.625% with epinephrine 1:300,000 (0.4 mL/kg) was injected after obtaining an elicited motor response at <0.4 mA of plantar flexion or inversion. Pinprick sensory assessments were performed at intervals by an observer unaware of the elicited motor response in the distal cutaneous distributions of the superficial peroneal nerve, deep peroneal nerve, posterior tibial nerve, and sural nerve. Motor block was assessed using foot (plantar flexion and dorsiflexion) movement and toe movement. A complete block was defined as sensory and motor loss in all distributions of the sciatic nerve within 25 minutes of local anesthetic injection. The optimal sensitivity and specificity of various cutoff times of sensory and motor testing were determined by receiver operating characteristic analysis. The area under the curves was compared for equivalence using nonparametric methods. The cutoff times were determined as the point of intersection of the lines of sensitivity and specificity. RESULTS: The elicited evoked motor response before sciatic nerve block was plantar flexion in 87 patients and inversion in 93. Eighty-eight of 93 patients (94.6%) who had an elicited motor response of inversion and 49 of 87 (55.7%) who had an elicited motor response of plantar flexion achieved complete sciatic nerve block at 25 minutes. Area under the curves were not different among testing paradigms. Receiver operating characteristic analysis identified optimal testing times of 4 minutes for the sural and 6 minutes for the posterior tibial nerve with an elicited motor response of inversion and 6 minutes with an elicited motor response of plantar flexion. No subject with an incomplete block achieved sural anesthesia by 10 minutes. CONCLUSION: Sural anesthesia assessed at the lateral heel and the lateral aspect of the foot and the fifth toe identified within 4 to 6 minutes demonstrated a similar posttest predictive value as anesthesia in the distributions of the posterior tibial and peroneal nerves or motor movement of the foot at later intervals. In addition, failure to achieve sural anesthesia within 10 minutes was predictive of block failure. PMID- 20007731 TI - Accuracy of methemoglobin detection by pulse CO-oximetry during hypoxia. AB - BACKGROUND: Methemoglobin in the blood cannot be detected by conventional pulse oximetry, although it can bias the oximeter's estimate (Spo2) of the true arterial functional oxygen saturation (Sao2). A recently introduced "Pulse CO Oximeter" (Masimo Rainbow SET(R) Radical-7 Pulse CO-Oximeter, Masimo Corp., Irvine, CA) is intended to additionally monitor noninvasively the fractional carboxyhemoglobin and methemoglobin content in blood. The purpose of our study was to determine whether hypoxia affects the new device's estimated methemoglobin reading accuracy, and whether the presence of methemoglobin impairs the ability of the Radical-7 and a conventional pulse oximeter (Nonin 9700, Nonin Medical Inc., Plymouth, MN) to detect decreases in Sao2. METHODS: Eight and 6 healthy adults were included in 2 study groups, respectively, each fitted with multiple sensors and a radial arterial catheter for blood sampling. In the first group, IV administration of approximately 300 mg sodium nitrite increased subjects' methemoglobin level to a 7% to 8% target and hypoxia was induced to different levels of Sao2 (70%-100%) by varying fractional inspired oxygen. In the second group, 15% methemoglobin at room air and 80% Sao2 were targeted. Pulse CO oximeter readings were compared with arterial blood values measured using a Radiometer multiwavelength hemoximeter. Pulse CO-oximeter methemoglobin reading performance was analyzed by observing the incidence of meaningful reading errors at the various hypoxia levels. This was used to determine the impact on predictive values for detecting methemoglobinemia. Spo2 reading bias, precision, and root mean square error were evaluated during conditions of elevated methemoglobin. RESULTS: Observations spanned 66.2% to 99% Sao2 and 0.6% to 14.4% methemoglobin over the 2 groups (170 blood draws). Masimo methemoglobin reading bias and precision over the full Sao2 span was 7.7% +/- 13.0%. Best accuracy was found in the 95% to 100% Sao2 range (1.9% +/- 2.5%), progressing to its worst in the 70% to 80% range (24.8% +/- 15.6%). Occurrence of methemoglobin readings in error >5% increased over each 5-point decrease in Sao2 (P < 0.05). Masimo Spo2 readings were biased -6.3% +/- 3.0% in the 95% to 100% Sao2 range with 4% to 8.3% methemoglobin. Both the Radical-7 and Nonin 9700 pulse oximeters accurately detected decreases in Sao(2) <90% with 4% to 15% methemoglobin, despite displaying low Spo2 readings when Sao2 was >95%. CONCLUSIONS: The Radical-7's methemoglobin readings become progressively more inaccurate as Sao2 decreases <95%, at times overestimating true values by 10% to 40%. Elevated methemoglobin causes the Spo2 readings to underestimate Sao2 similar to conventional 2 wavelength pulse oximeters at high saturation. Spo2 readings from both types of instruments continue to trend downward during the development of hypoxemia (Sao2 <90%) with methemoglobin levels up to 15%. PMID- 20007733 TI - Activation of spinal alpha-2 adrenoceptors, but not mu-opioid receptors, reduces the intrathecal N-methyl-D-aspartate-induced increase in spinal NR1 subunit phosphorylation and nociceptive behaviors in the rat. AB - BACKGROUND: A previous study from our laboratories showed that a significant reduction in spinal N-methyl-D-aspartate (NMDA) receptor NR1 subunit phosphorylation (pNR1) is associated with the antiallodynic effect produced by intrathecal (IT) injection of the alpha-2 adrenoceptor agonist, clonidine, in neuropathic rats. In this study, we determined whether the spontaneous pain and increased pNR1 expression induced by NMDA injection are reduced by IT injection of either clonidine or the mu-opioid receptor agonist, [D-Ala2, NMe-Phe4, Gly ol5]-enkephalin (DAMGO). METHODS: We examined the effect of clonidine (20 microg/rat) or DAMGO (1 microg/rat) injection on IT NMDA-induced spontaneous nociceptive behavior and pNR1 expression in the spinal dorsal horn. We also determined whether the effect of clonidine is mediated by alpha-2A or alpha-2C adrenoceptors. Finally, rat spinal cords were immunohistochemically processed for double staining of pNR1 and alpha-2A or alpha-2C adrenoceptors or mu-opioid receptors. RESULTS: The NMDA-induced increase in both pNR1 expression and nociceptive behavior was significantly reduced by IT clonidine but not DAMGO. This analgesic effect of clonidine was blocked by administration of either an alpha-2A (BRL44408, 30 microg/rat) or an alpha-2C (JP-1302, 50 microg/rat) adrenoceptor antagonist. In addition, immunocytochemistry revealed that spinal pNR1 immunoreactive cells co-contain alpha-2A and alpha-2C adrenoceptors. CONCLUSIONS: These results demonstrate that the IT NMDA-induced increase in pNR1 expression and nociceptive behavior is significantly reduced by activation of alpha-2 adrenoceptors, but not mu-opioid receptors, in the spinal cord dorsal horn. Furthermore, these findings suggest that the modulation of spinal NR1 phosphorylation is linked to the effect of IT clonidine on postsynaptic neuronal activity. PMID- 20007734 TI - The effects of nabilone on sleep in fibromyalgia: results of a randomized controlled trial. AB - BACKGROUND: Sleep disorders affect many patients with chronic pain conditions. Cannabis has been reported by several patient populations to help sleep. We evaluated the safety and efficacy of nabilone, a synthetic cannabinoid, on sleep disturbance in fibromyalgia (FM), a disease characterized by widespread chronic pain and insomnia. METHODS: We conducted a randomized, double-blind, active control, equivalency crossover trial to compare nabilone (0.5-1.0 mg before bedtime) to amitriptyline (10-20 mg before bedtime) in patients with FM with chronic insomnia. Subjects received each drug for 2 wk with a 2-wk washout period. The primary outcome was sleep quality, measured by the Insomnia Severity Index and the Leeds Sleep Evaluation Questionnaire. Secondary outcomes included pain, mood, quality of life, and adverse events (AEs). RESULTS: Thirty-one subjects were enrolled and 29 completed the trial (26 women, mean age 49.5 yr). Although sleep was improved by both amitriptyline and nabilone, nabilone was superior to amitriptyline (Insomnia Severity Index difference = 3.2; 95% confidence interval = 1.2-5.3). Nabilone was marginally better on the restfulness (Leeds Sleep Evaluation Questionnaire difference = 0.5 [0.0-1.0]) but not on wakefulness (difference = 0.3 [-0.2 to 0.8]). No effects on pain, mood, or quality of life were observed. AEs were mostly mild to moderate and were more frequent with nabilone. The most common AEs for nabilone were dizziness, nausea, and dry mouth. CONCLUSIONS: Nabilone is effective in improving sleep in patients with FM and is well tolerated. Low-dose nabilone given once daily at bedtime may be considered as an alternative to amitriptyline. Longer trials are needed to determine the duration of effect and to characterize long-term safety. PMID- 20007735 TI - Multidisciplinary approach to the challenge of hemostasis. AB - A multidisciplinary panel consisting of experts chosen by the 2 chairs of the group representing experts in anesthesiology, blood banking, hematology, critical care medicine, and various surgical disciplines (trauma, cardiac, pediatric, neurologic, obstetrics, and vascular) convened in January 2008 to discuss hemostasis and management of the bleeding patient across different clinical settings, with a focus on perioperative considerations. Although there are many ways to define hemostasis, one clinical definition would be control of bleeding without the occurrence of pathologic thrombotic events (i.e., when balance among procoagulant, anticoagulant, fibrinolytic, and antifibrinolytic activities is achieved). There are common hemostatic challenges that include lack of scientific evidence and standardized guidelines for the use of therapeutic drugs, need for reliable and rapid laboratory tools for measuring hemostasis, and individual variability. Clinically meaningful and accurate real-time laboratory data reflecting a patient's hemostatic status are needed to guide treatment decisions. Current available routine laboratory tests of hemostasis (e.g., platelet count, prothrombin time/international normalized ratio, and activated partial thromboplastin time) do not reflect the complexity of in vivo hemostasis and can mislead the clinician. Although point-of-care coagulation monitoring tests including measures of thromboelastography/elastometry provide insight into overall hemostatic status, they are time-consuming to perform, complex to interpret, and require trained personnel. There is a particular need to develop laboratory tests that can measure the effects of anticoagulant and antiplatelet agents for individual patients, predict bleeding complications, and guide therapy when and if treatment with blood products or pharmacologic drugs is required. Formation of an organization comprised of specialists who treat bleeding patients will foster multidisciplinary collaborations and promote discussions of the current state of hemostasis treatment and future priorities for hemostasis research. Controlled trials with clinically meaningful end points and suitable study populations, as well as observational studies, investigator-initiated studies, and large registry and database studies are essential to answer questions in hemostasis. Because of the complexities of maintaining hemostatic balance, advances in hemostasis research and continuing communication across specialties are required to improve patient care and outcomes. PMID- 20007736 TI - A comparison of propofol and dexmedetomidine for intravenous sedation: a randomized, crossover study of the effects on the central and autonomic nervous systems. AB - We compared, in volunteers, the effect of propofol (PROP) and dexmedetomidine (DEX) sedation on autonomic nervous activities and subjective feelings during psychological stresses. In a crossover design, 25 subjects received PROP and DEX titrated to a bispectral index value of 75 to 85. Heart rate, heart rate variability, and salivary alpha-amylase (objective indices) and a faces anxiety scale (subjective index) were assessed. Subjects were asked their preference between 2 sedatives. Objective indices showed similar changes in both groups. The faces anxiety scale decreased only in the PROP group and subjects preferred PROP. Propofol more effectively suppressed anxious feelings compared with DEX during sedation. PMID- 20007737 TI - A rare cause of recurrent hematuria in children: Nutcracker syndrome. AB - Nutcracker syndrome (NCS), the left renal vein hypertension due to compression between aorta and superior mesenteric artery, may present with hematuria, flank pain and proteinuria. We report a 6-year-old girl with episodic macroscopic hematuria who was diagnosed as NCS with Doppler ultrasound and 3D computerized tomography angiography. She was managed conservatively with spontaneous resolution of macroscopic hematuria. With this case we would like to emphasize that in children with hematuria and orthostatic proteinuria without an apparent cause, a great index of suspicion and appropriate imaging is necessary for the diagnosis of NCS. PMID- 20007738 TI - Dehydroepiandrosterone-induced proliferation of prostatic epithelial cell is mediated by NFKB via PI3K/AKT signaling pathway. AB - Dehydroepiandrosterone (DHEA) is an endogenous steroid that is metabolized to androgens and/or estrogens in the human prostate. DHEA levels decline with age, and use of DHEA supplements to retard the aging process is of unproved effectiveness and safety. In this study, rat ventral prostatic epithelial cells were used to determine whether DHEA-modulated proliferation and prostate-specific antigen (PSA listed as KLKB1 in the MGI Database) production were mediated via the androgen receptor (AR) and its potential mechanism. We demonstrated that proliferation of prostatic epithelial cells and increase of PSA expression induced by DHEA were neutralized by Casodex or Ar siRNA, two specific AR blockers. DHEA stimulated Nfkb DNA binding activity, with this effect being blunted by Casodex or Ar siRNA. Moreover, the inhibition of the phosphatidylinositol 3-kinase (PI3K)/AKT nullified the effects of DHEA on NFKB activation. These findings suggested that DHEA stimulated normal prostatic epithelial cell proliferation, and AR is involved in DHEA-induced PSA expression in normal prostatic epithelial cells. This stimulation effect induced by DHEA is mediated by the activation of NFKB via PI3K/AKT pathway. PMID- 20007739 TI - MARTA: a suite of Java-based tools for assigning taxonomic status to DNA sequences. AB - MOTIVATION: We have created a suite of Java-based software to better provide taxonomic assignments to DNA sequences. We anticipate that the program will be useful for protistologists, virologists, mycologists and other microbial ecologists. The program relies on NCBI utilities including the BLAST software and Taxonomy database and is easily manipulated at the command-line to specify a BLAST candidate's query-coverage or percent identity requirements; other options include the ability to set minimal consensus requirements (%) for each of the eight major taxonomic ranks (Domain, Kingdom, Phylum, ...) and whether to consider lower scoring candidates when the top-hit lacks taxonomic classification. PMID- 20007740 TI - Tmod: toolbox of motif discovery. AB - SUMMARY: Motif discovery is an important topic in computational transcriptional regulation studies. In the past decade, many researchers have contributed to the field and many de novo motif-finding tools have been developed, each may have a different strength. However, most of these tools do not have a user-friendly interface and their results are not easily comparable. We present a software called Toolbox of Motif Discovery (Tmod) for Windows operating systems. The current version of Tmod integrates 12 widely used motif discovery programs: MDscan, BioProspector, AlignACE, Gibbs Motif Sampler, MEME, CONSENSUS, MotifRegressor, GLAM, MotifSampler, SeSiMCMC, Weeder and YMF. Tmod provides a unified interface to ease the use of these programs and help users to understand the tuning parameters. It allows plug-in motif-finding programs to run either separately or in a batch mode with predetermined parameters, and provides a summary comprising of outputs from multiple programs. Tmod is developed in C++ with the support of Microsoft Foundation Classes and Cygwin. Tmod can also be easily expanded to include future algorithms. AVAILABILITY: Tmod is available for download at http://www.fas.harvard.edu/~junliu/Tmod/. PMID- 20007741 TI - QDD: a user-friendly program to select microsatellite markers and design primers from large sequencing projects. AB - SUMMARY: QDD is an open access program providing a user-friendly tool for microsatellite detection and primer design from large sets of DNA sequences. The program is designed to deal with all steps of treatment of raw sequences obtained from pyrosequencing of enriched DNA libraries, but it is also applicable to data obtained through other sequencing methods, using FASTA files as input. The following tasks are completed by QDD: tag sorting, adapter/vector removal, elimination of redundant sequences, detection of possible genomic multicopies (duplicated loci or transposable elements), stringent selection of target microsatellites and customizable primer design. It can treat up to one million sequences of a few hundred base pairs in the tag-sorting step, and up to 50,000 sequences in a single input file for the steps involving estimation of sequence similarity. AVAILABILITY: QDD is freely available under the GPL licence for Windows and Linux from the following web site: http://www.univ provence.fr/gsite/Local/egee/dir/meglecz/QDD.html. SUPPLEMENTARY INFORMATION: Supplementary data are available at Bioinformatics online. PMID- 20007742 TI - WebPARE: web-computing for inferring genetic or transcriptional interactions. AB - SUMMARY: Inferring genetic or transcriptional interactions, when done successfully, may provide insights into biological processes or biochemical pathways of interest. Unfortunately, most computational algorithms require a certain level of programming expertise. To provide a simple web interface for users to infer interactions from time course gene expression data, we present WebPARE, which is based on the pattern recognition algorithm (PARE). For expression data, in which each type of interaction (e.g. activator target) and the corresponding paired gene expression pattern are significantly associated, PARE uses a non-linear score to classify gene pairs of interest into a few subclasses of various time lags. In each subclass, PARE learns the parameters in the decision score using known interactions from biological experiments or published literature. Subsequently, the trained algorithm predicts interactions of a similar nature. Previously, PARE was shown to infer two sets of interactions in yeast successfully. Moreover, several predicted genetic interactions coincided with existing pathways; this indicates the potential of PARE in predicting partial pathway components. Given a list of gene pairs or genes of interest and expression data, WebPARE invokes PARE and outputs predicted interactions and their networks in directed graphs. PMID- 20007743 TI - Multiple organelle-targeting signals in the N-terminal portion of peroxisomal membrane protein PMP70. AB - Most membrane proteins are recognized by a signal recognition particle and are cotranslationally targeted to the endoplasmic reticulum (ER) membrane, whereas almost all peroxisomal membrane proteins are posttranslationally targeted to the destination. Here we examined organelle-targeting properties of the N-terminal portions of the peroxisomal isoform of the ABC transporter PMP70 (ABCD3) using enhanced green fluorescent protein (EGFP) fusion. When the N-terminal 80 amino acid residue (N80)-segment preceding transmembrane segment (TM) 1 was deleted and the TM1-TM2 region was fused to EGFP, the TM1 segment induced ER-targeting and integration in COS cells. When the N80-segment was fused to EGFP, the fusion protein was targeted to the outer mitochondrial membrane. When both the N80 segment and the following TM1-TM2 region were present, the fusion located exclusively to the peroxisome. The full-length PMP70 molecule was clearly located in the ER in the absence of the N80-segment, even when multiple peroxisome targeting signals were retained. We concluded that the TM1 segment possesses a sufficient ER-targeting function and that the N80-segment is critical for suppressing the ER-targeting function to allow the TM1-TM2 region to localize to the peroxisome. Cooperation of the organelle-targeting signals enables PMP70 to correctly target to peroxisomal membranes. PMID- 20007744 TI - Tobacco advertising through French TV in 2005: frequent illicit broadcasting; its impact on teenagers and young adults. AB - BACKGROUND: Sports sponsorship is one of the tobacco industry's main strategies to recruit new smokers among teenagers and young adults. METHODS: Monitoring Motor sports illicit broadcasting based on six channels in 2005; Dakar Rally (DR) and China Grand Prix impact evaluated with a one on one questionnaire administered on 12-24-year-old males and females (n = 805). RESULTS: 75,000 TV tobacco sponsoring appearances (90 h) were observed, total value: euro200.10(6); Mild Seven, Marlboro, West, Lucky Strike, Gauloises Blondes accounted for 92% appearances and 95% of euro values, with illegal broadcasting value worth euro19.10(6). A high interest in DR (71%) and Formula One (F1) (66%) was observed among males (versus females; P < 0.001), increasing with age (P < 0.05). Levels of spontaneous and assisted recall of cigarette brands were high among individuals interested in DR and F1, with better recall of brand names (versus non interested; P < 0.01); all individuals made correct associations between tobacco brands and car colors (P < 0.01). A non-significant higher percentage of daily smokers was observed among respondents interested in DR (34 versus 21%; P < 0.01) and F1 (30 versus 24%; P = 0.10). CONCLUSION: This high level of indirect advertising for tobacco through motor sport sponsoring confirms the urgent need for a worldwide absolute ban on tobacco advertising in motor sports. PMID- 20007745 TI - Preserving lessons learned in disease outbreaks and other emergency responses. AB - BACKGROUND: Public health departments often miss the opportunity to both learn from outbreaks and disasters and share any 'lessons learned' with other public health partners. These missed opportunities inhibit the public health system's ability to improve, change and adapt in an organized way. METHODS: In 2003, Western Pennsylvania experienced the largest documented hepatitis A outbreak in US history. The authors documented the key facets of both the outbreak and the response, and in the process developed improved methods for capturing the historical record in an efficient and comprehensive fashion. This process incorporates key aspects of oral history, along with typical public health parameters such as epidemic curves and environmental risk factors, and allows for creation of unique tools for documentation and sharing with diverse audiences. CONCLUSIONS: Learning from experience is an essential part of reducing mistakes, improving public health response, and the methods described herein show one way that 'lessons learned' can become a valuable teaching/training tool for students and practitioners. PMID- 20007746 TI - A comparison of fatigue correlates in rheumatoid arthritis and osteoarthritis: disparity in associations with disability, anxiety and sleep disturbance. AB - OBJECTIVES: To investigate correlates of fatigue among individuals with RA and OA, including mood, sleep, disease activity and radiographic damage. METHODS: Fatigue was assessed using the Multidimensional Assessment of Fatigue-Global Fatigue Index (MAF-GFI) in 103 patients with RA and 103 with OA. Sleep disturbance and pain were assessed using a visual analogue scale anxiety and depression using the Hospital Anxiety and Depression scale and disability using the HAQ. In the RA cohort, the disease activity score-28 joint count (DAS-28) and the Van der Heijde modified Sharp score were calculated, and in the OA cohort, the Kellgren-Lawrence score and the WOMAC score calculated. RESULTS: The MAF-GFI scores were higher in the OA cohort (P = 0.02). This was not significant after controlling for disability (P = 0.59). OA participants reported greater pain, disability, depression and sleeplessness than those with RA (all P < 0.01). The strongest correlates of fatigue in the RA cohort were depression (P < 0.001) and anxiety (P < 0.001). There was no significant association with pain (P = 0.43), DAS-28 (P = 0.07), HAQ (P = 0.10) or Sharp score (P = 0.78). In OA, the correlates of fatigue were older age (P = 0.02), sleep disturbance (P = 0.03), depression (P = 0.04), disability (P = 0.04) and lower CRP (P = 0.001). CONCLUSIONS: Fatigue is common and severe in both RA and OA. In RA, fatigue had no significant association with pain, disease activity, disability or erosions, but was associated with depression and anxiety. The disparity in correlates indicates that generalizing the experience of fatigue between OA and RA is not appropriate. Fatigue is an important domain in the assessment of disease impact. PMID- 20007747 TI - History of maltreatment and mental health problems in foster children: a review of the literature. AB - OBJECTIVE: Foster children often experience compromising situations such as neglect, physical abuse, or sexual abuse before out-of-home placement. This article aims to give a literature review related to the development and mental health of foster children with special consideration of trauma history. METHODS: A computer-based literature search was conducted in the databases Medline, PsycINFO, PSYNDEXplus, and SCOPUS. We determined a time frame from 1998 to 2009. RESULTS: The literature search resulted in 32 articles reporting empirical data about development and mental health in foster children. Very high rates of exposure to maltreatment, developmental delays and mental disorders were found. A broad spectrum of externalizing as well as internalizing symptoms and a high prevalence of comorbid mental disorders were found. CONCLUSIONS: Foster children exhibit a broad pattern of developmental problems and psychopathology. The etiology of these disorders is discussed in the context of multiple risk factors, especially that of persistent maltreatment. PMID- 20007749 TI - The Birth and Early Development of JASN. PMID- 20007750 TI - Protection from cancer in kidney transplant patients by gammadelta T cells: role of CMV infection? PMID- 20007751 TI - Cholecalciferol supplementation alters calcitriol-responsive monocyte proteins and decreases inflammatory cytokines in ESRD. AB - In vitro, monocyte 1alpha-hydroxylase converts 25-hydroxyvitamin D [25(OH)D] to 1,25-dihydroxyvitamin D to regulate local innate immune responses, but whether 25(OH)D repletion affects vitamin D-responsive monocyte pathways in vivo is unknown. Here, we identified seven patients who had 25(OH)D insufficiency and were undergoing long-term hemodialysis and assessed changes after cholecalciferol and paricalcitol therapies in both vitamin D-responsive proteins in circulating monocytes and serum levels of inflammatory cytokines. Cholecalciferol therapy increased serum 25(OH)D levels four-fold, monocyte vitamin D receptor expression three-fold, and 24-hydroxylase expression; therapy decreased monocyte 1alpha hydroxylase levels. The CD16(+) "inflammatory" monocyte subset responded to 25(OH)D repletion the most, demonstrating the greatest increase in vitamin D receptor expression after cholecalciferol. Cholecalciferol therapy reduced circulating levels of inflammatory cytokines, including IL-8, IL-6, and TNF. These data suggest that nutritional vitamin D therapy has a biologic effect on circulating monocytes and associated inflammatory markers in patients with ESRD. PMID- 20007752 TI - Carotid endarterectomy benefits patients with CKD and symptomatic high-grade stenosis. AB - Endarterectomy is generally recommended for symptomatic high-grade (70 to 99%) stenosis of the internal carotid artery, but whether this procedure is beneficial among patients with chronic kidney disease (CKD) is unknown. In this re-analysis of data from the North American Symptomatic Carotid Endarterectomy Trial, we included patients with symptomatic stenosis and either stage 3 CKD (n = 524) or preserved kidney function (n = 966; estimated GFR > or = 60). For medically treated patients with high-grade stenosis, risk for ipsilateral stroke at 2 yr was significantly higher in patients with CKD than in those with preserved renal function (31.6 versus 19.3%; P = 0.042); carotid endarterectomy significantly reduced this risk by 82 and 51%, respectively. To prevent one ipsilateral stroke, the number needed to treat by endarterectomy was four for patients with CKD and 10 for patients with preserved renal function. Compared with patients with preserved renal function, those with CKD had similar rates of perioperative stroke and death but higher rates of cardiac events. In conclusion, patients with stage 3 CKD and symptomatic high-grade carotid stenosis gain a large benefit in stroke risk reduction after endarterectomy. PMID- 20007753 TI - A contradictory role of A1 adenosine receptor in carbon tetrachloride- and bile duct ligation-induced liver fibrosis in mice. AB - Mice lacking A(1) adenosine receptors (A(1)AR) were thought to be protected from developing fatty liver; however, the contribution of A(1)AR to hepatic fibrosis has not been explored. Here we found that the expression of A(1)AR was decreased in fibrotic liver induced by chronic carbon tetrachloride (CCl(4)) but increased in that induced by bile duct ligation (BDL). Therefore, we examined whether A(1)AR contributes to hepatic fibrosis in CCl(4) and BDL animal models using A(1)AR knockout mice. Compared with wild-type (WT) mice, hepatic fibrosis resulting from chronic CCl(4) exposure was attenuated in A(1)AR(-/-) mice with markedly decreased collagen deposition and reduced hepatic stellate cell activation, whereas bile duct-ligated A(1)AR(-/-) mice displayed a significant increase in hepatic fibrosis. Hepatocyte damage was reduced in A(1)AR(-/-) mice after a single injection of CCl(4), with down-regulation of CYP2E1 and UCP2 gene expression in livers, which resulted in impaired liver sensitivity to CCl(4). However, BDL caused severe bile infarcts in livers of A(1)AR(-/-) mice, with significantly elevated levels of bile acid compared with those in WT mice. CCl(4) and BDL resulted in different expression patterns of genes involved in fibrogenesis in A(1)AR(-/-) mice. These results indicate that A(1)AR participates in the pathogenesis of hepatic fibrosis with a complex mechanism, and the effect of targeting adenosine and its receptors in the prevention of hepatic fibrosis should be cautiously evaluated. PMID- 20007754 TI - Sazetidine-A, a selective alpha4beta2 nicotinic receptor desensitizing agent and partial agonist, reduces nicotine self-administration in rats. AB - Adequate treatment of tobacco addiction remains problematic. Part of the problem with treatment is a poor understanding of the pharmacologic aspects of nicotine contributing to addiction. In addition to activating nicotinic acetylcholine receptors, nicotine also desensitizes them. It is currently not known how much of each of nicotine's actions contribute to its particular behavioral effects. Sazetidine-A (saz-A) is a novel nicotinic receptor-desensitizing agent and partial agonist with high selectivity for alpha4beta2 receptors. The current experiments were conducted to determine whether saz-A would reduce nicotine self administration in rats and to characterize its ancillary effects. Adult male Sprague-Dawley rats were allowed to self-administer nicotine. After initial food pellet training followed by 10 sessions of nicotine self-administration training, the rats were administered saz-A (0.1-3 mg/kg s.c.) or the saline vehicle in a repeated-measures counterbalanced design. Saz-A at the 3 mg/kg dose significantly decreased nicotine self-administration relative to performance of the same rats after saline injections. In a second study, long-term administration of this dose of sazetidine-A over the course of 10 sessions significantly reduced nicotine self-administration with no apparent diminution of effect. Saz-A in this dose range had only modest effects on locomotor activity, without any overall decrease in activity over a 1-h-long session. Saz-A significantly reduced food self administration, but this effect was smaller than its effect on nicotine self administration. Saz-A, which is a selective alpha4beta2-desensitizing agent and partial agonist, effectively reduces nicotine self-administration. This type of treatment holds promise for a new therapy to aid smoking cessation. PMID- 20007755 TI - Experience of 70-cm-long femoral tunnelled twin Tesio catheters for chronic haemodialysis. AB - BACKGROUND: Tunnelled femoral catheters with their tip in the lower inferior vena cava (IVC) are proposed only in few cases, but they often provide less than optimal blood flows and frequently have complications. The aim of this prospective observational study is to evaluate the use of 70-cm-long tunnelled cuffed femoral twin Tesio catheters with their tip in the upper IVC for haemodialysis. METHODS: Between May 2007 and May 2009, 25 tunnelled femoral catheters (fCVC) have been placed in 25 patients (77.7 +/- 10.8 years) with exhausted thoracic venous accesses or old patients with several comorbidities. Two 10 Fr carbothane 70-cm-long Tesio catheters with a Dacron cuff at 45 cm from the tip were placed in the femoral vein of each patient and then tunnelled; tips were in the upper third of the IVC. fCVCs were removed for either malfunction (Qb < 200 ml/min) or infection that did not resolve with antibiotics. RESULTS: Technical success of placement was 100%. The 6- and 12-month assisted primary patency rate were respectively 67 +/- 13% and 54 +/- 17%. The mean session Kt/V was 1.45 +/- 0.19, and the blood flow was 270 +/- 17 ml/min. Six fCVCs have been removed: three for infection, one for accidental damaging and two for the making of a different vascular access. The main complications were 2 catheter tip thrombi, 3 tunnel infections and 11 fCVC-related bacteraemia (1.77 episodes per 1000 CVC-days). CONCLUSION: The placement of twin fCVCs with their tip in the high IVC can provide an adequate dialysis and can be considered for patients with no remaining thoracic accesses. PMID- 20007756 TI - Effect of oral cromolyn sodium on CKD-associated pruritus and serum tryptase level: a double-blind placebo-controlled study. AB - BACKGROUND: Generalized pruritus is a significant complication in end-stage renal disease patients. The mechanism is unknown and most treatments are ineffective. This study is the first clinical trial designed to evaluate the effect of cromolyn sodium (CS) on renal itch. METHODS: Sixty-two haemodialysis (HD) patients with pruritus were enrolled into the study and were randomly assigned to receive CS or placebo (135 mg three times daily) for 8 weeks. Patients were asked to record the severity of their pruritus on each dialysis session on a visual analogue scale (VAS) during the 8 weeks of treatment and 4 weeks following discontinuation of treatment. Serum tryptase levels were determined at baseline, after 8 weeks of treatment and 4 weeks after discontinuation of treatment. RESULTS: Data were analysed in 21 patients in the CS group and 19 patients in the placebo group that completed the study. A significant difference was seen in the severity of pruritus between the two groups during the period of study. Level of pruritus decreased from 8.48 +/- 2.2 to 0.9 +/- 1.8 after 8 weeks of treatment with CS. Geometric mean of serum tryptase at baseline and 8 weeks after treatment were 21.3 and 19.5 ng/ml for the CS group and 18.03 and 18.2 ng/ml for the placebo group, respectively. Although the geometric mean of tryptase had decreased in the CS group, this decrease was not statistically significant (P = 0.214). CONCLUSION: CS can significantly reduce the severity of pruritus in HD patients, but this effect is not due to a decrease in serum tryptase level. PMID- 20007757 TI - Urothelial carcinoma transmission via kidney transplantation. AB - Transmission of urothelial carcinoma via solid organ transplant has never been reported in the literature to our knowledge. We report a case of transmission of this tumour to a kidney recipient. The donor was a 37-year-old woman, victim of a subarachnoid haemorrhage. The recipient was a 21-year-old girl, with a history of chronic kidney disease secondary to neurogenic bladder. This fatality has been rarely described in literature, but never with this histological type of cancer. Nowadays, with the expanded criteria for donation, older people are accepted as donor because of the shortage of organs. However, this may increase the likelihood of the number of cancer transmission. PMID- 20007758 TI - Mapping of a new locus for congenital anomalies of the kidney and urinary tract on chromosome 8q24. AB - BACKGROUND: Congenital anomalies of the kidney and urinary tract (CAKUT) account for the majority of end-stage renal disease in children (50%). Previous studies have mapped autosomal dominant loci for CAKUT. We here report a genome-wide search for linkage in a large pedigree of Somalian descent containing eight affected individuals with a non-syndromic form of CAKUT. METHODS: Clinical data and blood samples were obtained from a Somalian family with eight individuals with CAKUT including high-grade vesicoureteral reflux and unilateral renal agenesis. Total genome search for linkage was performed using a 50K SNP Affymetric DNA microarray. As neither parent is affected, the results of the SNP array were analysed under recessive models of inheritance, with and without the assumption of consanguinity. RESULTS: Using the non-consanguineous recessive model, a new gene locus (CAKUT1) for CAKUT was mapped to chromosome 8q24 with a significant maximum parametric Logarithm of the ODDs (LOD) score (LOD(max)) of 4.2. Recombinations were observed in two patients defining a critical genetic interval of 2.5 Mb physical distance flanked by markers SNP_A-1740062 and SNP_A 1653225. CONCLUSION: We have thus identified a new non-syndromic recessive gene locus for CAKUT (CAKUT1) on chromosome 8q24. The identification of the disease causing gene will provide further insights into the pathogenesis of urinary tract malformations and mechanisms of renal development. PMID- 20007759 TI - Absence of gadolinium deposits in the peritoneal membrane of patients with encapsulating peritoneal sclerosis. AB - Background. Encapsulating peritoneal sclerosis (EPS) is a severe complication of long-term peritoneal dialysis (PD) characterized by the development of an extensive fibrosis of the visceral peritoneum that may eventually lead to intestinal constriction. Its cause remains elusive. Nephrogenic systemic fibrosis (NSF), a disabling disease that can follow gadolinium-based contrast injection during magnetic resonance imaging, is characterized by systemic fibrosis of the skin, joints, liver, heart and vessels. Affected tissues are infiltrated by CD34+ and CD68+ fibroblasts. In the present study, we tested the hypothesis that EPS could have been triggered by a previous gadolinium injection. Methods. We performed histopathological analysis of the peritoneal membrane of two EPS and two control patients all exposed to long-term PD, including immunostaining with CD34 and CD68. The presence of gadolinium and other metals was also assessed by conventional and energy-filtered transmission electron microscopy. RESULTS: Numerous CD34+ and CD68+ cells were found in both the EPS and control patients within the vascular endothelium and in macrophages, respectively, but not in interstitial fibrocytes, as it could be expected in NSF. No trace of gadolinium deposits could be found in the four peritoneal samples; dispersed tiny iron inclusions were evidenced in the connective tissue of both EPS patients. CONCLUSIONS: These findings argue against the implication of gadolinium in the development of EPS in long-term PD patients. PMID- 20007760 TI - Prevalence of trimethoprim resistance genes in Escherichia coli isolates of human and animal origin in Lithuania. AB - A total of 456 non-repetitive Escherichia coli isolates from human clinical specimens (urinary, n=134; cervix, vagina and prostate, n=52; blood, pus and wounds, n=45), healthy animals (cattle, n=45; poultry, n=20) and diseased animals (cattle, n=53; swine, n=64; poultry, n=43) obtained in Lithuania during the period 2005-2008 were studied for trimethoprim (TMP) resistance and the prevalence of dfr genes. A TMP resistance rate in the range of 18-26 % respective to the origin was found in clinical isolates, 23-40 % in isolates from diseased animals and 9-20 % in isolates from healthy animals. Of 112 TMP-resistant isolates, 103 carried at least one of the six dfrA genes (dfrA1, dfrA5, dfrA8, dfrA12, dfrA14 and dfrA17) as determined by multiplex PCR and RFLP. The dfrA1 and dfrA17 genes were found most frequently in clinical isolates (17 and 19 isolates, respectively), whilst dfrA1 and dfrA14 genes dominated in isolates of animal origin (25 and 13 isolates, respectively). The dfrA5, dfrA12 and dfrA8 genes were detected at lower frequencies. The association with class 1/class 2 integrons was confirmed for 73-100 % of dfr genes found in most groups of isolates, except for the isolates from diseased swine. In this group, the majority of dfr-positive isolates (67 %, 8/12) carried dfrA8 (6/12) or dfrA14 genes (2/12) that were not associated with integrons. Non-integron location was also confirmed for the remaining dfrA8 genes (six clinical isolates and one isolate from diseased cattle) and for dfrA14 genes (two isolates from diseased cattle and swine each). All cassette-independent dfrA14 genes were found to be located within the strA gene. This study on the prevalence and distribution of TMP resistance genes among E. coli isolates of human and animal origin in Lithuania demonstrates that dfr genes are carried most frequently as gene cassettes within class 1 and/or class 2 integrons. However, TMP resistance in some of the isolates was found to be mediated by non-integron-associated dfrA8 and dfrA14 genes, indicating the existence of alternative sources for the spread of resistance. PMID- 20007761 TI - Classical RS1 and environmental RS1 elements in Vibrio cholerae O1 El Tor strains harbouring a tandem repeat of CTX prophage: revisiting Mozambique in 2005. AB - Currently, Vibrio cholerae O1 serogroup biotype El Tor strains producing classical type cholera toxin (altered strains or El Tor variants) are prevalent in Asia and in Mozambique. Mozambican strains collected in 2004 contained a tandem repeat of CTX prophage on the small chromosome and each CTX prophage harboured the classical rstR and classical ctxB. We found that the majority of the strains collected in 2005 in Mozambique contained extra elements on the large chromosome in addition to the tandem repeat of CTX prophage on the small chromosome. New type RS1 elements RS1(cla) and RS1(env), and a CTX(env) with rstR(env) and the classical ctxB were identified on the large chromosome of the Mozambican isolates collected in 2005. PMID- 20007762 TI - Diversity of staphylococcal cassette chromosome mec structures in coagulase negative staphylococci and relationship to drug resistance. AB - The objective of this study was to determine the distribution of staphylococcal cassette chromosome mec (SCCmec) elements in meticillin-resistant coagulase negative staphylococci (MR-CoNS) isolated from a tertiary-care hospital in Mexico and to examine the relationship to drug resistance. Fifty selected MR-CoNS isolates collected from catheters (n=15), blood (n=15), bone (n=9), bronchial lavage (n=2) and urine (n=2) and one isolate each from an abscess, cerebrospinal fluid, eye, pleural effusion, synovial fluid, tracheal aspirate and wound secretion were examined. Susceptibility testing was performed by the broth microdilution method. SCCmec types were determined by multiplex PCR and PFGE was carried out as described previously for Staphylococcus aureus. Among the MR-CoNS strains studied, the most frequently isolated species were Staphylococcus epidermidis (n=26) and Staphylococcus haemolyticus (n=13). Staphylococcus cohnii (n=5), Staphylococcus hominis (n=3), Staphylococcus sciuri (n=1), Staphylococcus pasteuri (n=1) and the recently described species Staphylococcus pettenkoferi (n=1) were also identified. The most frequent MR-CoNS genotype identified was SCCmec type IVa in S. epidermidis isolates, which also showed a high diversity in their PFGE patterns. A clone was found that amplified both SCCmec III and V elements in five isolates examined. The single MR S. pettenkoferi isolate harboured SCCmec type IVd and the single MR S. pasteuri isolate harboured SCCmec type I. The carriage of SCCmec type III was associated with resistance or intermediate resistance to meropenem (P <0.05). These results confirm the high prevalence of S. epidermidis SCCmec IVa and the high genetic diversity among MR CoNS strains. As far as is known, this is the first report describing the newly identified S. pettenkoferi possessing SCCmec IVd and S. pasteuri harbouring SCCmec type I. MR-CoNS harbouring SCCmec type III were found to be more resistant to meropenem. PMID- 20007763 TI - Isolation of a bacteriophage specific for CS7-expressing strains of enterotoxigenic Escherichia coli. AB - Enterotoxigenic Escherichia coli (ETEC) is the most common bacterial cause of childhood diarrhoea in Bangladesh. Among the virulence factors of ETEC, toxins and colonization factors (CFs) play a major role in pathogenesis. Unlike Vibrio cholerae, the relationship between ETEC and ETEC-specific phages is poorly understood and the possible role of ETEC phages in the evolution of ETEC strains in the environment is yet to be established. This study was designed specifically to isolate phages that are specific for ETEC virulence factors. Among the 49 phages isolated from 12 different surface water samples, 13 were tested against 211 ETEC strains collected from clinical and environmental sources. One phage, designated IMM-001, showed a significant specificity towards CS7 CF as it attacked all the CS7-expressing ETEC. Electron microscopic analyses showed that the isolated phage possessed an isomeric hexagonal head and a long filamentous tail. An antibody blocking method and phage neutralization assay confirmed that CS7 pilus is required for the phage infection process, indicating the role of CS7 fimbrial protein as a potential receptor for IMM-001. In summary, this study showed the presence of a lytic phage in environmental water that is specific for the CS7 CF of ETEC. PMID- 20007764 TI - Typing of the pilus-protein-encoding FCT region and biofilm formation as novel parameters in epidemiological investigations of Streptococcus pyogenes isolates from various infection sites. AB - Streptococcus pyogenes is an important human pathogen for which an association between infection site and selected epidemiological or functional markers has previously been suggested. However, the studies involved often used strains with an insufficiently defined clinical background and laboratory history. Thus, the major goal of the present study was to investigate these relationships in 183 prospectively collected, well-defined, low-passage isolates from a North-East German centre for tertiary care. For each isolate the clinical background (91 respiratory, 71 skin and 21 invasive isolates) and antibiotic-resistance pattern was recorded. All isolates were classified according to their emm type, antibiotic-resistance and PFGE pattern ( SmaI restriction analysis of genomic DNA). As novel discriminatory methods we performed a PCR-based typing of the pilus-protein-encoding FCT region (FCT) and biofilm-formation phenotyping in various culture media. Forty-one isolates were found to be resistant to at least one of the tested antibiotics. emm typing revealed emm28, emm12 , emm1, emm4, emm89 and emm2 as the most frequent types in our collection. The novel FCT typing showed isolates encoding FCT types 4 and 2 to be the most common. Overall 113 strains with unique combinations of emm and FCT types, antibiotic-resistance and PFGE patterns were identified. The majority of all isolates revealed an association of biofilm-formation capacity with growth media. Comparing all results for potential associations, no correlation could be established between the anatomical site of isolation and the emm or the FCT type. There was no relationship between biofilm formation and emm type, antibiotic-resistance or PFGE patterns. However, a novel association between biofilm formation and FCT type became obvious among strains from our collection. PMID- 20007765 TI - A comparative analysis of molecular markers for the detection and identification of Borrelia spirochaetes in Ixodes ricinus. AB - Borrelia burgdorferi sensu lato, carried by Ixodes ticks, is one of the most significant human pathogens, causing Lyme disease. As there is no standardized PCR method for detection and identification of spirochaete DNA, we carried out a comparative analysis using a set of complementary primers for three regions in the genomic DNA of these bacteria (genes fla and rrs and the non-coding rrs-rrlA region). DNA extracted from 579 Ixodes ricinus ticks was subjected to nested PCR. DNA of the examined spirochaetes was detected in 43 (7.4 %) lysates when the fla gene was used as a molecular marker, in 7 (1.2 %) lysates when using primers complementary to the rrs gene, and in 12 (2.1 %) lysates using primers complementary to the non-coding rrs-rrlA sequence. RFLP analysis based on the fla gene helped identify species from the B. burgdorferi sensu lato complex (B. burgdorferi sensu stricto, Borrelia afzelii, Borrelia garinii, Borrelia valaisiana), detect co-infections, and also identify Borrelia miyamotoi. Therefore, the fla gene is the most sensitive and specific molecular marker for the detection and identification of Borrelia spirochaetes in I. ricinus. PMID- 20007766 TI - A backward progression of attentional effects in the ventral stream. AB - The visual processing of behaviorally relevant stimuli is enhanced through top down attentional feedback. One possibility is that feedback targets early visual areas first and the attentional enhancement builds up at progressively later stages of the visual hierarchy. An alternative possibility is that the feedback targets the higher-order areas first and the attentional effects are communicated "backward" to early visual areas. Here, we compared the magnitude and latency of attentional enhancement of firing rates in V1, V2, and V4 in the same animals performing the same task. We found a reverse order of attentional effects, such that attentional enhancement was larger and earlier in V4 and smaller and later in V1, with intermediate results in V2. These results suggest that attentional mechanisms operate via feedback from higher-order areas to lower-order ones. PMID- 20007767 TI - Inflammation and adipose tissue macrophages in lipodystrophic mice. AB - Lipodystrophy and obesity are opposites in terms of a deficiency versus excess of adipose tissue mass, yet these conditions are accompanied by similar metabolic consequences, including insulin resistance, dyslipidemia, hepatic steatosis, and increased risk for diabetes and atherosclerosis. Hepatic and myocellular steatosis likely contribute to metabolic dysregulation in both states. Inflammation and macrophage infiltration into adipose tissue also appear to participate in the pathogenesis of obesity-induced insulin resistance, but their contributions to lipodystrophy-induced insulin resistance have not been evaluated. We used aP2-nSREBP-1c transgenic (Tg) mice, an established model of lipodystrophy, to ask this question. Circulating cytokine elevations suggested systemic inflammation but even more dramatic was the number of infiltrating macrophages in all white and brown adipose tissue depots of the Tg mice; in contrast, there was no evidence of inflammatory infiltrates or responses in any other tissue including liver. Despite there being overt evidence of adipose tissue inflammation, antiinflammatory strategies including salicylate treatment and genetic suppression of myeloid NF-kappaB signaling that correct insulin resistance in obesity were ineffective in the lipodystrophic mice. We further showed that adipose tissue macrophages (ATMs) in lipodystrophy and obesity are very different in terms of activation state, gene expression patterns, and response to lipopolysaccharide. Although ATMs are even more abundant in lipodystrophy than in obesity, they have distinct phenotypes and likely roles in tissue remodeling, but do not appear to be involved in the pathogenesis of insulin resistance. PMID- 20007769 TI - Network analyses structure genetic diversity in independent genetic worlds. AB - DNA flows between chromosomes and mobile elements, following rules that are poorly understood. This limited knowledge is partly explained by the limits of current approaches to study the structure and evolution of genetic diversity. Network analyses of 119,381 homologous DNA families, sampled from 111 cellular genomes and from 165,529 phage, plasmid, and environmental virome sequences, offer challenging insights. Our results support a disconnected yet highly structured network of genetic diversity, revealing the existence of multiple "genetic worlds." These divides define multiple isolated groups of DNA vehicles drawing on distinct gene pools. Mathematical studies of the centralities of these worlds' subnetworks demonstrate that plasmids, not viruses, were key vectors of genetic exchange between bacterial chromosomes, both recently and in the past. Furthermore, network methodology introduces new ways of quantifying current sampling of genetic diversity. PMID- 20007770 TI - The role of seawater endocytosis in the biomineralization process in calcareous foraminifera. AB - Foraminifera are unicellular organisms that inhabit the oceans in various ecosystems. The majority of the foraminifera precipitate calcitic shells and are among the major CaCO(3) producers in the oceans. They comprise an important component of the global carbon cycle and also provide valuable paleoceanographic information based on the relative abundance of stable isotopes and trace elements (proxies) in their shells. Understanding the biomineralization processes in foraminifera is important for predicting their calcification response to ocean acidification and for reliable interpretation of the paleoceanographic proxies. Most models of biomineralization invoke the involvement of membrane ion transporters (channels and pumps) in the delivery of Ca(2+) and other ions to the calcification site. Here we show, in contrast, that in the benthic foraminiferan Amphistegina lobifera, (a shallow water species), transport of seawater via fluid phase endocytosis may account for most of the ions supplied to the calcification site. During their intracellular passage the seawater vacuoles undergo alkalization that elevates the CO(3)(2-) concentration and further enhances their calcifying potential. This mechanism of biomineralization may explain why many calcareous foraminifera can be good recorders of paleoceanographic conditions. It may also explain the sensitivity to ocean acidification that was observed in several planktonic and benthic species. PMID- 20007771 TI - miR319a targeting of TCP4 is critical for petal growth and development in Arabidopsis. AB - In a genetic screen in a drnl-2 background, we isolated a loss-of-function allele in miR319a (miR319a(129)). Previously, miR319a has been postulated to play a role in leaf development based on the dramatic curled-leaf phenotype of plants that ectopically express miR319a (jaw-D). miR319a(129) mutants exhibit defects in petal and stamen development; petals are narrow and short, and stamens exhibit defects in anther development. The miR319a(129) loss-of-function allele contains a single-base change in the middle of the encoded miRNA, which reduces the ability of miR319a to recognize targets. Analysis of the expression patterns of the three members of the miR319 gene family (miR319a, miR319b, and miR319c) indicates that these genes have largely non-overlapping expression patterns suggesting that these genes have distinct developmental functions. miR319a functions by regulating the TCP transcription factors TCP2, TCP3, TCP4, TCP10, and TCP24; the level of RNA expression of these TCP genes is down-regulated in jaw-D and elevated in miR319a(129). Several lines of evidence demonstrate that TCP4 is a key target of miR319a. First, the tcp4(soj6) mutant, which contains a mutation in the TCP4 miRNA-binding site complementary to the miR319a(129) mutation, suppresses the flower phenotype of miR319a(129). Second, expression of wild-type TCP4 in petals and stamens (i.e., AP3:TCP4) has no effect on flower development; by contrast, a miRNA-resistant version of TCP4, when expressed in petals and stamens (i.e., pAP3:mTCP4) causes these organs not to develop. Surprisingly, when AP3:TCP4 is present in a miR319a(129) background, petal and stamen development is severely disrupted, suggesting that proper regulation by miR319a of TCP4 is critical in these floral organs. PMID- 20007768 TI - Large-scale patterns in biodiversity of microbial eukaryotes from the abyssal sea floor. AB - Eukaryotic microbial life at abyssal depths remains "uncharted territory" in eukaryotic microbiology. No phylogenetic surveys have focused on the largest benthic environment on this planet, the abyssal plains. Moreover, knowledge of the spatial patterns of deep-sea community structure is scanty, and what little is known originates primarily from morphology-based studies of foraminiferans. Here we report on the great phylogenetic diversity of microbial eukaryotic communities of all 3 abyssal plains of the southeastern Atlantic Ocean--the Angola, Cape, and Guinea Abyssal Plains--from depths of 5,000 m. A high percentage of retrieved clones had no close representatives in genetic databases. Many clones were affiliated with parasitic species. Furthermore, differences between the communities of the Cape Abyssal Plain and the other 2 abyssal plains point to environmental gradients apparently shaping community structure at the landscape level. On a regional scale, local species diversity showed much less variation. Our study provides insight into the community composition of microbial eukaryotes on larger scales from the wide abyssal sea floor realm and marks a direction for more detailed future studies aimed at improving our understanding of deep-sea microbes at the community and ecosystem levels, as well as the ecological principles at play. PMID- 20007772 TI - Functional enhancement and protection of dopaminergic terminals by RAB3B overexpression. AB - In Parkinson's disease (PD), dopaminergic (DA) neurons in the substantia nigra (SN, A9) are particularly vulnerable, compared to adjacent DA neurons within the ventral tegmental area (VTA, A10). Here, we show that in rat and human, one RAB3 isoform, RAB3B, has higher expression levels in A10 compared to A9 neurons. RAB3 is a monomeric GTPase protein that is highly enriched in synaptic vesicles and is involved in synaptic vesicle trafficking and synaptic transmission, disturbances of which have been implicated in several neurodegenerative diseases, including PD. These findings prompted us to further investigate the biology and neuroprotective capacity of RAB3B both in vitro and in vivo. RAB3B overexpression in human dopaminergic BE (2)-M17 cells increased neurotransmitter content, [(3)H] dopamine uptake, and levels of presynaptic proteins. AAV-mediated RAB3B overexpression in A9 DA neurons of the rat SN increased striatal dopamine content, number and size of synaptic vesicles, and levels of the presynaptic proteins, confirming in vitro findings. Measurement of extracellular DOPAC, a dopamine metabolite, following l-DOPA injection supported a role for RAB3B in enhancing the dopamine storage capacity of synaptic terminals. RAB3B overexpression in BE (2)-M17 cells was protective against toxins that simulate aspects of PD in vitro, including an oxidative stressor 6-hydroxydopamine (6 OHDA) and a proteasome inhibitor MG-132. Furthermore, RAB3B overexpression in rat SN both protected A9 DA neurons and resulted in behavioral improvement in a 6 OHDA retrograde lesion model of PD. These results suggest that RAB3B improves dopamine handling and storage capacity at presynaptic terminals, and confers protection to vulnerable DA neurons. PMID- 20007773 TI - Differences in human and chimpanzee gene expression patterns define an evolving network of transcription factors in brain. AB - Humans differ from other primates by marked differences in cognitive abilities and a significantly larger brain. These differences correlate with metabolic changes, as evidenced by the relative up-regulation of energy-related genes and metabolites in human brain. While the mechanisms underlying these evolutionary changes have not been elucidated, altered activities of key transcription factors (TFs) could play a pivotal role. To assess this possibility, we analyzed microarray data from five tissues from humans and chimpanzees. We identified 90 TF genes with significantly different expression levels in human and chimpanzee brain among which the rapidly evolving KRAB-zinc finger genes are markedly over represented. The differentially expressed TFs cluster within a robust regulatory network consisting of two distinct but interlinked modules, one strongly associated with energy metabolism functions, and the other with transcription, vesicular transport, and ubiquitination. Our results suggest that concerted changes in a relatively small number of interacting TFs may coordinate major gene expression differences in human and chimpanzee brain. PMID- 20007774 TI - The DM domain protein DMRT1 is a dose-sensitive regulator of fetal germ cell proliferation and pluripotency. AB - Dmrt1 (doublesex and mab-3 related transcription factor 1) is a conserved transcriptional regulator of male differentiation required for testicular development in vertebrates. Here, we show that in mice of the 129Sv strain, loss of Dmrt1 causes a high incidence of teratomas, whereas these tumors do not form in Dmrt1 mutant C57BL/6J mice. Conditional gene targeting indicates that Dmrt1 is required in fetal germ cells but not in Sertoli cells to prevent teratoma formation. Mutant 129Sv germ cells undergo apparently normal differentiation up to embryonic day 13.5 (E13.5), but some cells fail to arrest mitosis and ectopically express pluripotency markers. Expression analysis and chromatin immunoprecipitation identified DMRT1 target genes, whose missexpression may underlie teratoma formation. DMRT1 indirectly activates the GDNF coreceptor Ret, and it directly represses the pluripotency regulator Sox2. Analysis of human germ cell tumors reveals similar gene expression changes correlated to DMRT1 levels. Dmrt1 behaves genetically as a dose-sensitive tumor suppressor gene in 129Sv mice, and natural variation in Dmrt1 activity can confer teratoma susceptibility. This work reveals a genetic link between testicular dysgenesis, pluripotency regulation, and teratoma susceptibility that is highly sensitive to genetic background and to gene dosage. PMID- 20007775 TI - Alterations of the Notch pathway in lung cancer. AB - Notch signaling regulates cell specification and homeostasis of stem cell compartments, and it is counteracted by the cell fate determinant Numb. Both Numb and Notch have been implicated in human tumors. Here, we show that Notch signaling is altered in approximately one third of non-small-cell lung carcinomas (NSCLCs), which are the leading cause of cancer-related deaths: in approximately 30% of NSCLCs, loss of Numb expression leads to increased Notch activity, while in a smaller fraction of cases (around 10%), gain-of-function mutations of the NOTCH-1 gene are present. Activation of Notch correlates with poor clinical outcomes in NSCLC patients without TP53 mutations. Finally, primary epithelial cell cultures, derived from NSCLC harboring constitutive activation of the Notch pathway, are selectively killed by inhibitors of Notch (gamma-secretase inhibitors), showing that the proliferative advantage of these tumors is dependent upon Notch signaling. Our results show that the deregulation of the Notch pathway is a relatively frequent event in NSCLCs and suggest that it might represent a possible target for molecular therapies in these tumors. PMID- 20007776 TI - High resilience in the Yamal-Nenets social-ecological system, West Siberian Arctic, Russia. AB - Tundra ecosystems are vulnerable to hydrocarbon development, in part because small-scale, low-intensity disturbances can affect vegetation, permafrost soils, and wildlife out of proportion to their spatial extent. Scaling up to include human residents, tightly integrated arctic social-ecological systems (SESs) are believed similarly susceptible to industrial impacts and climate change. In contrast to northern Alaska and Canada, most terrestrial and aquatic components of West Siberian oil and gas fields are seasonally exploited by migratory herders, hunters, fishers, and domesticated reindeer (Rangifer tarandus L.). Despite anthropogenic fragmentation and transformation of a large proportion of the environment, recent socioeconomic upheaval, and pronounced climate warming, we find the Yamal-Nenets SES highly resilient according to a few key measures. We detail the remarkable extent to which the system has successfully reorganized in response to recent shocks and evaluate the limits of the system's capacity to respond. Our analytical approach combines quantitative methods with participant observation to understand the overall effects of rapid land use and climate change at the level of the entire Yamal system, detect thresholds crossed using surrogates, and identify potential traps. Institutional constraints and drivers were as important as the documented ecological changes. Particularly crucial to success is the unfettered movement of people and animals in space and time, which allows them to alternately avoid or exploit a wide range of natural and anthropogenic habitats. However, expansion of infrastructure, concomitant terrestrial and freshwater ecosystem degradation, climate change, and a massive influx of workers underway present a looming threat to future resilience. PMID- 20007777 TI - Mitaplatin, a potent fusion of cisplatin and the orphan drug dichloroacetate. AB - The unique glycolytic metabolism of most solid tumors, known as the Warburg effect, is associated with resistance to apoptosis that enables cancer cells to survive. Dichloroacetate (DCA) is an anticancer agent that can reverse the Warburg effect by inhibiting a key enzyme in cancer cells, pyruvate dehydrogenase kinase (PDK), that is required for the process. DCA is currently not approved for cancer treatment in the USA. Here, we present the synthesis, characterization, and anticancer properties of c,t,c-[Pt(NH(3))(2)(O(2)CHCl(2))(2)Cl(2)], mitaplatin, in which two DCA units are appended to the axial positions of a six coordinate Pt(IV) center. The negative intracellular redox potential reduces the platinum to release cisplatin, a Pt(II) compound, and two equivalents of DCA. By a unique mechanism, mitaplatin thereby attacks both nuclear DNA with cisplatin and mitochondria with DCA selectively in cancer cells. The cytotoxicity of mitaplatin in a variety of cancer cell lines equals or exceeds that of all known Pt(IV) compounds and is comparable to that of cisplatin. Mitaplatin alters the mitochondrial membrane potential gradient (Deltapsi(m)) of cancer cells, promoting apoptosis by releasing cytochrome c and translocating apoptosis inducing factor from mitochondria to the nucleus. Cisplatin formed upon cellular reduction of mitaplatin enters the nucleus and targets DNA to form 1,2 intrastrand d(GpG) cross-links characteristic of its own potency as an anticancer drug. These properties of mitaplatin are manifest in its ability to selectively kill cancer cells cocultured with normal fibroblasts and to partially overcome cisplatin resistance. PMID- 20007778 TI - Cross-modal cueing of attention alters appearance and early cortical processing of visual stimuli. AB - The question of whether attention makes sensory impressions appear more intense has been a matter of debate for over a century. Recent psychophysical studies have reported that attention increases apparent contrast of visual stimuli, but the issue continues to be debated. We obtained converging neurophysiological evidence from human observers as they judged the relative contrast of visual stimuli presented to the left and right visual fields following a lateralized auditory cue. Cross-modal cueing of attention boosted the apparent contrast of the visual target in association with an enlarged neural response in the contralateral visual cortex that began within 100 ms after target onset. The magnitude of the enhanced neural response was positively correlated with perceptual reports of the cued target being higher in contrast. The results suggest that attention increases the perceived contrast of visual stimuli by boosting early sensory processing in the visual cortex. PMID- 20007779 TI - Uncoupling of sustained MAMP receptor signaling from early outputs in an Arabidopsis endoplasmic reticulum glucosidase II allele. AB - Recognition of microbe-associated molecular patterns (MAMPs), conserved structures typical of a microbial class, triggers immune responses in eukaryotes. This is accompanied by a diverse set of physiological responses that are thought to enhance defense activity in plants. However, the extent and mechanisms by which MAMP-induced events contribute to host immunity are poorly understood. Here we reveal Arabidopsis priority in sweet life4 (psl4) and psl5 mutants that are insensitive to the bacterial elongation factor (EF)-Tu epitope elf18 but responsive to flagellin epitope flg22. PSL4 and PSL5, respectively, identify beta and alpha-subunits of endoplasmic reticulum-resident glucosidase II, which is essential for stable accumulation and quality control of the elf18 receptor EFR but not the flg22 receptor FLS2. We notice that EFR signaling is partially and differentially impaired without a significant decrease of the receptor steady state levels in 2 weakly dysfunctional gIIalpha alleles, designated psl5-1 and rsw3. Remarkably, rsw3 plants exhibit marked supersusceptibility against a virulent bacterial phytopathogen despite nearly intact coactivation of MAPKs, reactive oxygen species, ethylene biosynthesis, and callose deposition in response to elf18, demonstrating that these signaling outputs alone are insufficient to mount effective immunity. However, rsw3 plants fail to maintain high transcript levels of defense-promoting WRKY, PR1, and PR2 genes at late time points (4 to 24 h) after elf18 elicitation. This points to an unexpected separation between initial and sustained activation of EFR-mediated signaling in the absence of proper glucosidase II-mediated endoplasmic reticulum quality control. Our findings strongly suggest the importance of sustained MAMP receptor signaling as a key step in the establishment of robust immunity. PMID- 20007780 TI - Distinguishing influence-based contagion from homophily-driven diffusion in dynamic networks. AB - Node characteristics and behaviors are often correlated with the structure of social networks over time. While evidence of this type of assortative mixing and temporal clustering of behaviors among linked nodes is used to support claims of peer influence and social contagion in networks, homophily may also explain such evidence. Here we develop a dynamic matched sample estimation framework to distinguish influence and homophily effects in dynamic networks, and we apply this framework to a global instant messaging network of 27.4 million users, using data on the day-by-day adoption of a mobile service application and users' longitudinal behavioral, demographic, and geographic data. We find that previous methods overestimate peer influence in product adoption decisions in this network by 300-700%, and that homophily explains >50% of the perceived behavioral contagion. These findings and methods are essential to both our understanding of the mechanisms that drive contagions in networks and our knowledge of how to propagate or combat them in domains as diverse as epidemiology, marketing, development economics, and public health. PMID- 20007783 TI - Recollection versus strength as the primary determinant of hippocampal engagement at retrieval. AB - We examined whether hippocampal activity in recognition relates to the strength of the memory or to recollective experience, a subject of considerable current debate. Participants studied word pairs and then made two successive recognition decisions on each item: first on the uncued target and then on the target presented with the studied cue word. We compared recollection and familiarity patterns of activation in fMRI for these decisions. Critically, our analyses attempted in two ways to equate perceived memory strength while varying the associative information available. First, activity for targets judged familiar before cueing was contrasted with activity for the same items in the second decision as a function of whether the targets converted to recollection or remained familiar when the context cues were provided. We found increased hippocampal activity following cueing only with recollective conversion. Second, we investigated whether hippocampal activity was modulated by the rated familiarity strength of cued items or whether it increased uniquely in recollection. Hippocampal activation was not modulated parametrically by familiarity strength and recollected items were associated with greater activity relative to highly familiar items. Together, our results support the notion that it is recollection of context, rather than memory strength, that underlies hippocampal engagement at retrieval. PMID- 20007781 TI - Specific apoptosis induction by the dual PI3K/mTor inhibitor NVP-BEZ235 in HER2 amplified and PIK3CA mutant breast cancer cells. AB - NVP-BEZ235 is a dual PI3K/mTOR inhibitor currently in phase I clinical trials. We profiled this compound against a panel of breast tumor cell lines to identify the patient populations that would benefit from such treatment. In this setting, NVP BEZ235 selectively induced cell death in cell lines presenting either HER2 amplification and/or PIK3CA mutation, but not in cell lines with PTEN loss of function or KRAS mutations, for which resistance could be attributed, in part to ERK pathway activity. An in depth analysis of death markers revealed that the cell death observed upon NVP-BEZ235 treatment could be recapitulated with other PI3K inhibitors and that this event is linked to active PARP cleavage indicative of an apoptotic process. Moreover, the effect seemed to be partly independent of the caspase-9 executioner and mitochondrial activated caspases, suggesting an alternate route for apoptosis induction by PI3K inhibitors. Overall, this study will provide guidance for patient stratification for forthcoming breast cancer phase II trials for NVP-BEZ235. PMID- 20007782 TI - A designer ligand specific for Kv1.3 channels from a scorpion neurotoxin-based library. AB - Venomous animals immobilize prey using protein toxins that act on ion channels and other targets of biological importance. Broad use of toxins for biomedical research, diagnosis, and therapy has been limited by inadequate target discrimination, for example, among ion channel subtypes. Here, a synthetic toxin is produced by a new strategy to be specific for human Kv1.3 channels, critical regulators of immune T cells. A phage display library of 11,200 de novo proteins is designed using the alpha-KTx scaffold of 31 scorpion toxin sequences known or predicted to bind to potassium channels. Mokatoxin-1 (moka1) is isolated by affinity selection on purified target. Moka1 blocks Kv1.3 at nanomolar levels that do not inhibit Kv1.1, Kv1.2, or KCa1.1. As a result, moka1 suppresses CD3/28 induced cytokine secretion by T cells without cross-reactive gastrointestinal hyperactivity. The 3D structure of moka1 rationalizes its specificity and validates the engineering approach, revealing a unique interaction surface supported on an alpha-KTx scaffold. This scaffold-based/target-biased strategy overcomes many obstacles to production of selective toxins. PMID- 20007784 TI - Error-prone translesion replication of damaged DNA suppresses skin carcinogenesis by controlling inflammatory hyperplasia. AB - The induction of skin cancer involves both mutagenic and proliferative responses of the epidermis to ultraviolet (UV) light. It is believed that tumor initiation requires the mutagenic replication of damaged DNA by translesion synthesis (TLS) pathways. The mechanistic basis for the induction of proliferation, providing tumor promotion, is poorly understood. Here, we have investigated the role of TLS in the initiation and promotion of skin carcinogenesis, using a sensitive nucleotide excision repair-deficient mouse model that carries a hypomorphic allele of the error-prone TLS gene Rev1. Despite a defect in UV-induced mutagenesis, skin carcinogenesis was accelerated in these mice. This paradoxical phenotype was caused by the induction of inflammatory hyperplasia of the mutant skin that provides strong tumor promotion. The induction of hyperplasia was associated with mild and transient replicational stress of the UV-damaged genome, triggering DNA damage signaling and senescence. The concomitant expression of Interleukin-6 (IL-6) is in agreement with an executive role for IL-6 and possibly other cytokines in the autocrine induction of senescence and the paracrine induction of inflammatory hyperplasia. In conclusion, error-prone TLS suppresses tumor-promoting activities of UV light, thereby controlling skin carcinogenesis. PMID- 20007785 TI - Two-component protein-engineered physical hydrogels for cell encapsulation. AB - Current protocols to encapsulate cells within physical hydrogels require substantial changes in environmental conditions (pH, temperature, or ionic strength) to initiate gelation. These conditions can be detrimental to cells and are often difficult to reproduce, therefore complicating their use in clinical settings. We report the development of a two-component, molecular-recognition gelation strategy that enables cell encapsulation without environmental triggers. Instead, the two components, which contain multiple repeats of WW and proline rich peptide domains, undergo a sol-gel phase transition upon simple mixing and hetero-assembly of the peptide domains. We term these materials mixing-induced, two-component hydrogels. Our results demonstrate use of the WW and proline-rich domains in protein-engineered materials and expand the library of peptides successfully designed into engineered proteins. Because both of these association domains are normally found intracellularly, their molecular recognition is not disrupted by the presence of additional biomolecules in the extracellular milieu, thereby enabling reproducible encapsulation of multiple cell types, including PC 12 neuronal-like cells, human umbilical vein endothelial cells, and murine adult neural stem cells. Precise variations in the molecular-level design of the two components including (i) the frequency of repeated association domains per chain and (ii) the association energy between domains enable tailoring of the hydrogel viscoelasticity to achieve plateau shear moduli ranging from approximately 9 to 50 Pa. Because of the transient physical crosslinks that form between association domains, these hydrogels are shear-thinning, injectable, and self-healing. Neural stem cells encapsulated in the hydrogels form stable three-dimensional cultures that continue to self-renew, differentiate, and sprout extended neurites. PMID- 20007786 TI - The atomic structure of baculovirus polyhedra reveals the independent emergence of infectious crystals in DNA and RNA viruses. AB - Baculoviruses are ubiquitous insect viruses well known for their use as bioinsecticides, gene therapy vectors, and protein expression systems. Overexpression of recombinant proteins in insect cell culture utilizes the strong promoter of the polyhedrin gene. In infected larvae, the polyhedrin protein forms robust intracellular crystals called polyhedra, which protect encased virions for prolonged periods in the environment. Polyhedra are produced by two unrelated families of insect viruses, baculoviruses and cypoviruses. The atomic structure of cypovirus polyhedra revealed an intricate packing of trimers, which are interconnected by a projecting N-terminal helical arm of the polyhedrin molecule. Baculovirus and cypovirus polyhedra share nearly identical lattices, and the N terminal region of the otherwise unrelated baculovirus polyhedrin protein sequence is also predicted to be alpha-helical. These results suggest homology between the proteins and a common structural basis for viral polyhedra. Here, we present the 2.2-A structure of baculovirus polyhedra determined by x-ray crystallography from microcrystals produced in vivo. We show that the underlying molecular organization is, in fact, very different. Although both polyhedra have nearly identical unit cell dimensions and share I23 symmetry, the polyhedrin molecules are structurally unrelated and pack differently in the crystals. In particular, disulfide bonds and domain-swapped N-terminal domains stabilize the building blocks of baculovirus polyhedra and interlocking C-terminal arms join unit cells together. We show that the N-terminal projecting helical arms have different structural roles in baculovirus and cypovirus polyhedra and conclude that there is no structural evidence for a common evolutionary origin for both classes of polyhedra. PMID- 20007787 TI - Sequential formation of ion pairs during activation of a sodium channel voltage sensor. AB - Electrical signaling in biology depends upon a unique electromechanical transduction process mediated by the S4 segments of voltage-gated ion channels. These transmembrane segments are driven outward by the force of the electric field on positively charged amino acid residues termed "gating charges," which are positioned at three-residue intervals in the S4 transmembrane segment, and this movement is coupled to opening of the pore. Here, we use the disulfide locking method to demonstrate sequential ion pair formation between the fourth gating charge in the S4 segment (R4) and two acidic residues in the S2 segment during activation. R4 interacts first with E70 at the intracellular end of the S2 segment and then with D60 near the extracellular end. Analysis with the Rosetta Membrane method reveals the 3-D structures of the gating pore as these ion pairs are formed sequentially to catalyze the S4 transmembrane movement required for voltage-dependent activation. Our results directly demonstrate sequential ion pair formation that is an essential feature of the sliding helix model of voltage sensor function but is not compatible with the other widely discussed gating models. PMID- 20007788 TI - Magnesium-aspartate-based crystallization switch inspired from shell molt of crustacean. AB - Many animals such as crustacean periodically undergo cyclic molt of the exoskeleton. During this process, amorphous calcium mineral phases are biologically stabilized by magnesium and are reserved for the subsequent rapid formation of new shell tissue. However, it is a mystery how living organisms can regulate the transition of the precursor phases precisely. We reveal that the shell mineralization from the magnesium stabilized precursors is associated with the presence of Asp-rich proteins. It is suggested that a cooperative effect of magnesium and Asp-rich compound can result into a crystallization switch in biomineralization. Our in vitro experiments confirm that magnesium increases the lifetime of amorphous calcium carbonate and calcium phosphate in solution so that the crystallization can be temporarily switched off. Although Asp monomer alone inhibits the crystallization of pure amorphous calcium minerals, it actually reduces the stability of the magnesium-stabilized precursors to switch on the transformation from the amorphous to crystallized phases. These modification effects on crystallization kinetics can be understood by an Asp-enhanced magnesium desolvation model. The interesting magnesium-Asp-based switch is a biologically inspired lesson from nature, which can be developed into an advanced strategy to control material fabrications. PMID- 20007789 TI - Absence of geochemical evidence for an impact event at the Bolling Allerod/Younger Dryas transition. AB - High concentrations of iridium have been reported in terrestrial sediments dated at 12.9 ka and are interpreted to support an extraterrestrial impact event as the cause of the observed extinction in the Rancholabrean fauna, changes in the Paleoindian cultures, and the onset of the Younger Dryas cooling [Firestone RB, et al. (2007) Proc Natl Acad Sci USA 104:16016-16021]. Here, we report platinum group element (PGE: Os, Ir, Ru, Rh, Pt, Pd), gold (Au) concentrations, and (187)Os/(188)Os ratios in time-equivalent terrestrial, lacustrine, and marine sections to seek robust evidence of an extraterrestrial contribution. First, our results do not reproduce the previously reported elevated Ir concentrations. Second, (187)Os/(188)Os isotopic ratios in the sediment layers investigated are similar to average crustal values, indicating the absence of a significant meteoritic Os contribution to these sediments. Third, no PGE anomalies distinct from crustal signatures are present in the marine record in either the Gulf of California (DSDP 480, Guaymas Basin) or the Cariaco Basin (ODP 1002C). Our data show no evidence of an extraterrestrial (ET)-PGE enrichment anomaly in any of the investigated depositional settings investigated across North America and in one section in Belgium. The lack of a clear ET-PGE signature in this sample suite is inconsistent with the impact of a large chondritic projectile at the Bolling Allerod/Younger Dryas transition. PMID- 20007791 TI - Awareness and anaesthesia: think dose, think data. PMID- 20007792 TI - Reduced clearance of rocuronium and sugammadex in patients with severe to end stage renal failure: a pharmacokinetic study. AB - BACKGROUND: Sugammadex is a selective relaxant binding agent designed to encapsulate the neuromuscular blocking agent, rocuronium. The sugammadex rocuronium complex is eliminated by the kidneys. This trial investigated the pharmacokinetics (PKs) of sugammadex and rocuronium in patients with renal failure and healthy controls. METHODS: Fifteen ASA class II-III renal patients [creatinine clearance (CL(CR)) <30 ml min(-1)] and 15 ASA I-II controls (CL(CR) > or =80 ml min(-1)) were included. After induction of anaesthesia, a single i.v. dose of rocuronium 0.6 mg kg(-1) was given, followed by a single i.v. dose of sugammadex 2.0 mg kg(-1) at reappearance of the second twitch of the train-of four response. Plasma concentrations of rocuronium and sugammadex were estimated and PK variables determined using non-compartmental analyses. Percentages of sugammadex and rocuronium excreted in the urine were measured. RESULTS: PK data were obtained from 26 patients. Mean total plasma clearance (CL) of sugammadex was 5.5 ml min(-1) in renal patients and 95.2 ml min(-1) in controls (P<0.05). Rocuronium CL was 41.8 ml min(-1) in renal patients and 167 ml min(-1) in controls (P<0.05). The median amount of sugammadex and rocuronium excreted in the urine over 72 h in renal patients was 29% and 4%, respectively, and 73% and 42% over 24 h in controls. CONCLUSIONS: Large differences in the PKs of sugammadex and rocuronium between patients with renal failure and healthy controls were observed. The effect of renal impairment on the PK variables of rocuronium was less than with sugammadex. Urinary excretion of both drugs was reduced in renal patients. PMID- 20007793 TI - Mallampati class changes during pregnancy, labour, and after delivery: can these be predicted? AB - BACKGROUND: An increase in Mallampati class is associated with difficult laryngoscopy in obstetrics. The goal of our study was to determine the changes in Mallampati class before, during, and after labour, and to identify predictive factors of the changes. METHODS: Mallampati class was evaluated at four time intervals in 87 pregnant patients: during the 8th month of pregnancy (T(1)), placement of epidural catheter (T(2)), 20 min after delivery (T(3)), and 48 h after delivery (T(4)). Factors such as gestational weight gain, duration of first and second stages of labour, and i.v. fluids administered during labour were evaluated for their predictive value. Mallampati classes 3 and 4 were compared for each time interval. Logistic regression was used to test the association between each factor and Mallampati class evolution. RESULTS: Mallampati class did not change for 37% of patients. The proportion of patients falling into Mallampati classes 3 and 4 at the various times of assessment were: T(1), 10.3%; T(2), 36.8%; T(3), 51.7%; and T(4), 20.7%. The differences in percentages were all significant (P<0.01). None of the evaluated factors was predictive. CONCLUSIONS: The incidence of Mallampati classes 3 and 4 increases during labour compared with the pre-labour period, and these changes are not fully reversed by 48 h after delivery. This work confirms the absolute necessity of examining the airway before anaesthetic management in obstetric patients. PMID- 20007794 TI - Combined acetaminophen and ibuprofen for pain relief after oral surgery in adults: a randomized controlled trial. AB - BACKGROUND: Acetaminophen is often used with a non-steriodal anti-inflammatory drug for acute pain. Hitherto, these drugs have had to be given separately, typically at different time intervals. Maxigesic tablets combine acetaminophen and ibuprofen in clinically appropriate doses to simplify administration and dosage regimen. We compared this combination with each of the constituent drugs for the relief of pain after extraction of third molar teeth. METHODS: Adults (more than 16 yr) having one or more wisdom teeth removed under general or local anaesthesia were instructed to take two tablets before operation, then two tablets every 6 h for up to 48 h of: (i) a combination of acetaminophen 500 mg and ibuprofen 150 mg per tablet (Maxigesic); (ii) acetaminophen 500 mg per tablet alone; or (iii) ibuprofen 150 mg per tablet alone. The primary outcome measure was the area under the curve (AUC) of the 100 mm visual analogue scale pain measurements taken for up to 48 h after surgery, divided by time, at rest and on activity. Pharmacokinetic data were collected in a subset of patients. RESULTS: The mean (sem) time-corrected AUC on rest and activity, respectively, were: combination group 22.3 (3.2) and 28.4 (3.4); acetaminophen group 33.0 (3.1) and 40.4 (3.3); and ibuprofen group 34.8 (3.2) and 40.2 (3.4); P<0.01 for each of the four comparisons of combination vs constituent drug. There was no pharmacokinetic interaction between acetaminophen and ibuprofen administered together. CONCLUSIONS: Maxigesic tablets provide superior pain relief after oral surgery to acetaminophen or ibuprofen alone. PMID- 20007795 TI - Relationship of inspiratory and expiratory times to upper airway resistance during pulsatile needle cricothyrotomy ventilation with generic delivery circuit. AB - BACKGROUND: Narrow-bore cricothyrotomy retains a clinical role, due to the availability of its component equipment in acute clinical environments, ease of assembly, and operator preference. However, due to infrequent use, there is a need to model this for research and teaching. We present mathematical and laboratory models. METHODS: Using electrical analogy, we mathematically modelled a generic cannula cricothyrotomy circuit, relating inspiratory and expiratory times to the upper airway resistance (R(u)). We constructed a laboratory model to support our mathematical model. The simulated lung is a smooth-bore tube on a tilting table. The upper airway is simulated by 20 G cannulae. Inspiratory and expiratory times for the water meniscus to travel a preset distance (corresponding to tidal volume) were measured and plotted against the number of cannula. RESULTS: From the mathematical model, inspiratory time increases hyperbolically with decreasing R(u), such that there is a minimum R(u) beyond which most of the fresh gas flow leaks out without inflating the chest. Conversely, as R(u) increases, inspiratory time decreases to a plateau. Expiratory time is limited by respiratory factors at low R(u) and by the resistance of the transtracheal expiratory pathway at high R(u), producing a sigmoid-shaped expiratory curve. The experimental results seem consistent with these predictions, although direct theory-experiment mapping is problematic because of the difficulty in assigning a single value to the dynamically changing upper airway resistance. CONCLUSIONS: We can exploit the contrasting changes in inspiratory and expiratory times with the upper airway resistance to optimize conditions for emergent cannula cricothyrotomy ventilation. PMID- 20007796 TI - Application of a ventilator care bundle and outcome. PMID- 20007797 TI - Simulation in anaesthesia training. PMID- 20007798 TI - Cardiopulmonary arrest in pregnancy. PMID- 20007799 TI - Continuous flow positive airway pressure generator in critically ill patients. PMID- 20007800 TI - Anaesthetic management of a patient for femoral correction using a new multi axial fixation system. PMID- 20007801 TI - McGrath videolaryngoscope for awake tracheal intubation in a patient with severe ankylosing spondylitis. PMID- 20007802 TI - Lung isolation with a new Y-shaped endobronchial blocking device, the EZ-Blocker. PMID- 20007803 TI - FLT3-mutant allelic burden and clinical status are predictive of response to FLT3 inhibitors in AML. AB - We examined 6 different FMS-like tyrosine kinase-3 (FLT3) inhibitors (lestaurtinib, midostaurin, AC220, KW-2449, sorafenib, and sunitinib) for potency against mutant and wild-type FLT3, as well as for cytotoxic effect against a series of primary blast samples obtained from patients with acute myeloid leukemia (AML) harboring internal tandem duplication (FLT3/ITD) mutations. We found that inhibition of FLT3 autophosphorylation in a FLT3/ITD specimen does not always induce cell death, suggesting that some FLT3/ITD AML may not be addicted to FLT3 signaling. Relapsed samples and samples with a high mutant allelic burden were more likely to be responsive to cytotoxicity from FLT3 inhibition compared with the samples obtained at diagnosis or those with a low mutant allelic burden. These FLT3 inhibitors varied to a considerable degree in their selectivity for FLT3, and this selectivity influenced the cytotoxic effect. These results have important implications for the potential therapeutic use of FLT3 inhibitors in that patients with newly diagnosed FLT3-mutant AML might be less likely to respond clinically to highly selective FLT3 inhibition. PMID- 20007804 TI - Ras-induced reactive oxygen species promote growth factor-independent proliferation in human CD34+ hematopoietic progenitor cells. AB - Excessive production of reactive oxygen species (ROS) is a feature of human malignancy and is often triggered by activation of oncogenes such as activated Ras. ROS act as second messengers and can influence a variety of cellular process including growth factor responses and cell survival. We have examined the contribution of ROS production to the effects of N-Ras(G12D) and H-Ras(G12V) on normal human CD34(+) progenitor cells. Activated Ras strongly up-regulated the production of both superoxide and hydrogen peroxide through the stimulation of NADPH oxidase (NOX) activity, without affecting the expression of endogenous antioxidants or the production of mitochondrially derived ROS. Activated Ras also promoted both the survival and the growth factor-independent proliferation of CD34(+) cells. Using oxidase inhibitors and antioxidants, we found that excessive ROS production by these cells did not contribute to their enhanced survival; rather, ROS promoted their growth factor-independent proliferation. Although Ras induced ROS production specifically activated the p38(MAPK) oxidative stress response, this failed to induce expression of the cell-cycle inhibitor, p16(INK4A); instead, ROS promoted the expression of D cyclins. These data are the first to show that excessive ROS production in the context of oncogene activation can promote proliferative responses in normal human hematopoietic progenitor cells. PMID- 20007805 TI - A CD8 T cell-intrinsic role for the calcineurin-NFAT pathway for tolerance induction in vivo. AB - Previous studies have indicated that blockade of signaling through the T-cell receptor (TCR)/calcineurin/nuclear factor of activated T cells (NFAT) pathway impairs transplantation tolerance induced with anti-CD154 antibody. By using an allogeneic bone marrow transplantation model, we examined the role of the TCR/calcineurin/NFAT pathway for tolerance induction with anti-CD154. Calcineurin blockade by cyclosporine A led to a failure of CD8 but not CD4 tolerance, and experiments in NFAT1(-/-) mice replicated this effect. Studies in thymectomized mice demonstrated that blockade of the calcineurin/NFAT pathway after bone marrow transplantation led to a failure of peripheral CD8 tolerance. Moreover, CD8 adoptive transfer studies demonstrated that NFAT1 is cell-intrinsically required for peripheral CD8 tolerance. NFAT1 deficiency did not impair CD8 T-cell up regulation of PD1, which is required for CD8 tolerance in this model. NFAT1 has previously been shown to have a role in CD4 cells for anergy induction and for programming CD4 cells to become regulatory cells. By generating mice lacking NFAT1 in CD4 but not CD8 cells, we demonstrate that NFAT1 is neither required for CD4 tolerance induction nor for their regulatory function on CD8 T cells. Thus, our study reveals a CD8 T cell-intrinsic NFAT1 requirement for CD8 tolerance in vivo. PMID- 20007806 TI - Emergence of BCR-ABL-specific cytotoxic T cells in the bone marrow of patients with Ph+ acute lymphoblastic leukemia during long-term imatinib mesylate treatment. AB - Imatinib mesylate has been demonstrated to allow the emergence of T cells directed against chronic myeloid leukemia cells. A total of 10 Philadelphia chromosome-positive acute lymphoblastic leukemia patients receiving high-dose imatinib mesylate maintenance underwent long-term immunological monitoring (range, 2-65 months) of (p190)BCR-ABL-specific T cells in the bone marrow and peripheral blood. (p190)BCR-ABL-specific T lymphocytes were detected in all patients, more frequently in bone marrow than in peripheral blood samples (67% vs 25%, P < .01) and resulted significantly associated with lower minimal residual disease values (P < .001), whereas absent at leukemia relapse. Specific T cells were mainly effector memory CD8(+) and CD4(+) T cells, producing interferon gamma, tumor necrosis factor-alpha, and interleukin-2 (median percentage of positive cells: 3.34, 3.04, and 3.58, respectively). Cytotoxic subsets able to lyse BCR-ABL-positive leukemia blasts also were detectable. Whether these autologous (p190)BCR-ABL-specific T cells may be detectable under other tyrosine kinase inhibitors, expanded ex vivo, and exploited for immunotherapy remains to be addressed. PMID- 20007807 TI - Free HTLV-1 induces TLR7-dependent innate immune response and TRAIL relocalization in killer plasmacytoid dendritic cells. AB - A recent report demonstrated that free human T-cell leukemia virus 1 (HTLV-1) could infect plasmacytoid dendritic cells (pDCs). The major role of pDCs is to secrete massive levels of interferon-alpha (IFN-alpha) upon virus exposure; however, the induction of IFN-alpha by HTLV-1 remains unknown. We demonstrate here that cell-free HTLV-1 generated a pDC innate immune response by producing massive levels of IFN-alpha that were inhibited by anti-HTLV-1 antibodies. HTLV-1 induced costimulatory molecules and rapid expression of the apoptotic ligand tumor necrosis factor-related apoptosis-inducing ligand (TRAIL). Furthermore, HTLV-1 stimulated pDC-induced apoptosis of CD4(+) T cells expressing DR5, transforming pDCs into IFN-producing killer pDCs. We also observed that an endosomal acidification inhibitor and a Toll-like receptor-7 (TLR7)-specific blocker drastically inhibited pDC response to HTLV-1. Three-dimensional microscopy analysis revealed that unstimulated pDCs were "dormant" IFN-producing killer pDCs with high levels of intracellular TRAIL that could be rapidly mobilized to the surface in response to TLR7 activation. Inhibition of viral degradation in endosomes by chloroquine maintained viral integrity, allowing virus detection by 3-dimensional microscopy. We demonstrate that pDCs respond to cell-free HTLV-1 by producing high levels of IFN-alpha and by mobilizing TRAIL on cell surface after TLR7 triggering. This is the first demonstration of an innate immune response induced by free HTLV-1. PMID- 20007808 TI - A murine model of severe immune thrombocytopenia is induced by antibody- and CD8+ T cell-mediated responses that are differentially sensitive to therapy. AB - Immune thrombocytopenia (ITP) is a bleeding disorder characterized by antibody opsonized platelets being prematurely destroyed in the spleen, although some patients with ITP may have a cell-mediated form of thrombocytopenia. Although several animal models of ITP have been developed, few mimic primary chronic ITP nor have any shown cell-mediated platelet destruction. To create this type of model, splenocytes from CD61 knockout mice immunized against CD61(+) platelets were transferred into severe combined immunodeficient (SCID) (CD61(+)) mouse recipients, and their platelet counts and phenotypes were observed. As few as 5 x 10(4) splenocytes induced a significant thrombocytopenia and bleeding mortality (80%) in recipients within 3 weeks after transfer. Depletion of lymphocyte subsets before transfer showed that the splenocyte's ability to induce thrombocytopenia and bleeding completely depended on CD4(+) T helper cells and that both CD19(+) B cell (antibody)- and CD8(+) T cell (cell)-mediated effector mechanisms were responsible. Treatment of the SCID mouse recipients with intravenous gamma-globulins raised platelet counts and completely prevented bleeding mortality induced by antibody-mediated effector mechanisms but did not affect cell-mediated disease. This novel model not only shows both antibody- and cell-mediated ITP and bleeding but also suggests that these 2 effector mechanisms have a differential response to therapy. PMID- 20007809 TI - Intravenous PEG-asparaginase during remission induction in children and adolescents with newly diagnosed acute lymphoblastic leukemia. AB - Over the past several decades, L-asparaginase, an important component of therapy for acute lymphoblastic leukemia (ALL), has typically been administered intramuscularly rather than intravenously in North America because of concerns regarding anaphylaxis. We evaluated the feasibility of giving polyethylene glycosylated (PEG)-asparaginase, the polyethylene glycol conjugate of Escherichia coli L-asparaginase, by intravenous infusion in children with ALL. Between 2005 and 2007, 197 patients (age, 1-17 years) were enrolled on Dana-Farber Cancer Institute ALL Consortium Protocol 05-01 and received a single dose of intravenous PEG-asparaginase (2500 IU/m(2)) over 1 hour during remission induction. Serum asparaginase activity more than 0.1 IU/mL was detected in 95%, 88%, and 7% of patients at 11, 18, and 25 days after dosing, respectively. Toxicities included allergy (1.5%), venous thrombosis (2%), and pancreatitis (4.6%). We conclude that intravenous administration of PEG-asparaginase is tolerable in children with ALL, and potentially therapeutic enzyme activity is maintained for at least 2 weeks after a single dose in most patients. This trial was registered at www.clinicaltrials.gov as #NCT00400946. PMID- 20007810 TI - Crystal structure and collagen-binding site of immune inhibitory receptor LAIR-1: unexpected implications for collagen binding by platelet receptor GPVI. AB - Leukocyte-associated immunoglobulin-like receptor-1 (LAIR-1), one of the most widely spread immune receptors, attenuates immune cell activation when bound to specific sites in collagen. The collagen-binding domain of LAIR-1 is homologous to that of glycoprotein VI (GPVI), a collagen receptor crucial for platelet activation. Because LAIR-1 and GPVI also display overlapping collagen-binding specificities, a common structural basis for collagen recognition would appear likely. Therefore, it is crucial to gain insight into the molecular interaction of both receptors with their ligand to prevent unwanted cross-reactions during therapeutic intervention. We determined the crystal structure of LAIR-1 and mapped its collagen-binding site by nuclear magnetic resonance (NMR) titrations and mutagenesis. Our data identify R59, E61, and W109 as key residues for collagen interaction. These residues are strictly conserved in LAIR-1 and GPVI alike; however, they are located outside the previously proposed GPVI collagen binding site. Our data provide evidence for an unanticipated mechanism of collagen recognition common to LAIR-1 and GPVI. This fundamental insight will contribute to the exploration of specific means of intervention in collagen induced signaling in immunity and hemostasis. PMID- 20007811 TI - TGFBIp: more than meets the eye? PMID- 20007812 TI - CD44: target for antiangiogenesis therapy. PMID- 20007813 TI - Fetal calf serum contains heat-stable nucleases that degrade neutrophil extracellular traps. PMID- 20007814 TI - Health-related quality of life, smoking and carotid atherosclerosis in white British women with systemic lupus erythematosus. AB - We tested the hypothesis that carotid atherosclerosis in systemic lupus erythematosus (SLE) is associated with poor health-related quality of life (HRQOL), which is independent of any association with traditional risk factors (TRFs), lifestyle and socioeconomic factors. Women with SLE completed the RAND Medical Outcome Study 36-Item Short-Form Health Survey version 1 (MOS SF-36). B mode Doppler examination of the carotid arteries determined the presence of atherosclerotic plaque. The association between carotid plaque and HRQOL domains was analysed using logistic regression models with sequential adjustments for age, TRFs, education level and employment status. We studied 181 women, 47 (26%) of whom had carotid plaque. Carotid plaque was significantly associated with lower levels of physical functioning (p = 0.047), vitality (p = 0.04), role emotional (p = 0.04) and mental health subscales (p = 0.01) and lower mental component summary score (MCS) (p = 0.03). These associations were no longer significant after adjustment for age and TRFs, especially smoking. Smokers had lower physical functioning, vitality and mental health and more bodily pain. The association between carotid plaque and HRQOL was not independent of TRFs and smoking was a key mediator of the associations found. Poor HRQOL in smokers will need addressing as part of any smoking cessation strategies in SLE patients. PMID- 20007815 TI - Bone marrow infection caused by Mycobacterium avium complex in a patient with systemic lupus erythematosus. AB - Systemic lupus erythematosus (SLE) is a disease with wide range of signs and symptoms. SLE patients have increased infective diathesis, and infections are a very important cause of death in these patients. Infections can sometimes mimic the signs and symptoms of SLE. Thus, it is important to recognize that infection can induce a lupus flare-up or can be difficult to distinguish from a lupus flare up. We describe a 36-year-old female patient with SLE, who presented with skin lesions and pancytopenia, and clinical manifestations similar to a flare-up of SLE. Bone marrow examination revealed infection with Mycobacterium avium complex (MAC). The patient had no history or clinical evidence of pulmonary involvement. This patient is the first case of invasive bone marrow MAC infection in SLE. With this unique case, we would like to emphasize that SLE patients can also be infected by non-tuberculous mycobacteria, and that bone marrow examination for tuberculosis as well as for non-tuberculosis mycobacteria should be considered in SLE patients with refractory pancytopenia. PMID- 20007816 TI - Lytic bone lesions in lupus-associated myelofibrosis. AB - Myelofibrosis is associated with numerous causes other than hematologic malignant neoplasms, including autoimmune diseases. We describe a 44-year-old woman who suffered bone lytic lesions with extramedullary haematopoiesis in the setting of myelofibrosis associated with systemic lupus erythematosus. PMID- 20007817 TI - Safe time limits of aortic cross-clamping and cardiopulmonary bypass in adult cardiac surgery. AB - OBJECTIVES: We evaluated the impact of aortic cross-clamping time (XCT) and cardiopulmonary bypass time (CPBT) on the immediate and late outcome after adult cardiac surgery and attempted to identify their safe time limits. METHODS: This study includes 3280 patients who underwent adult cardiac surgery of various complexities. Myocardial protection was achieved with tepid continuous antegrade/retrograde blood cardioplegia. RESULTS: Receiver operating characteristics (ROC) curve analysis showed that XCT (area under the curve, AUC: 0.66), CPBT (AUC: 0.73) and CPBT with unclamped aorta (AUC: 0.77) were significantly associated with 30-day postoperative mortality. XCT of increasing 30-minute intervals (Odds Ratio (OR) 1.21, 95%C.I. 1.01-1.52) and CPBT of increasing 30-minute intervals (OR 1.47, 95%C.I. 1.27-1.71) were independent predictors of 30-day mortality. The best cutoff value for XCT was 150 min (30-day death: 1.8% vs. 12.2%, adjusted OR 3.07, 95%C.I. 1.48-6.39, accuracy 91.5%) and for CPBT 240 min (30-day death: 1.9% vs. 31.5%, adjusted OR 8.78, 95%C.I. 4.64 16.61, accuracy 96.0%). These parameters were significantly associated also with postoperative morbidity, particularly with postoperative stroke. CONCLUSIONS: XCT and CPBT are predictors of immediate postoperative morbidity and mortality. In our experience, cardiac procedures with CPBT<240 min and XCT<150 min were associated with a rather low risk of immediate postoperative adverse events independently of the complexity of surgery patient's operative risk. PMID- 20007818 TI - Transitions between care settings at the end of life in the Netherlands: results from a nationwide study. AB - Multiple transitions between care settings in the last phase of life could jeopardize continuity of care and overall end-of-life patient care. Using a mortality follow-back study, we examined the nature and prevalence of transitions between Dutch care settings in the last 3 months of life, and identified potential characteristics associated with them. During the 2-year study period, 690 registered patients died 'totally expectedly and non-suddenly'. These made 709 transitions in the last 3 months, which involved a hospital two times out of three, and covered 43 distinct care trajectories. The most frequent trajectory was home-to-hospital (48%). Forty-six percent experienced one or more transitions in their last month of life. Male gender, multi-morbidities, and absence of GP awareness of a patient's wish for place of death were associated with having a transition in the last 30 days of life; age of < or = 85 years, having an infection and the absence of a palliative-centred treatment goal were associated with terminal hospitalization for > or = 7 days. Although the majority of the 'totally expected and non-sudden' deaths occurred at home, transitions to hospitals were relatively frequent. To minimize abrupt or frequent transitions just before death, timely recognition of the palliative phase of dying is important. PMID- 20007819 TI - Measuring quality of life in pediatric palliative care: challenges and potential solutions. AB - Annually, about 500,000 children are coping with life-limiting illnesses in the USA. Integrated pediatric palliative care program could benefit some of these children by improving their health-related quality of life (HRQOL). To measure the effect of pediatric palliative care programs on HRQOL, a valid and reliable tool must be identified. This study aimed to validate the psychometric properties of a generic HRQOL instrument, the Pediatric Quality of Life 4.0, for children with life-limiting illnesses. Analyses were conducted using telephone survey data collected from 266 parents whose Medicaid-enrolled children had life-limiting illnesses. Results of the analyses suggest the Pediatric Quality of Life 4.0 does not have valid psychometric properties for measuring HRQOL within this population. Our study documents several challenges in using the generic instrument to measure HRQOL in pediatric palliative care setting. We point out future directions to refine or develop HRQOL instruments for this population of vulnerable children. PMID- 20007820 TI - Inviting the absent members: examining how caregivers' participation affects hospice team communication. AB - This paper is a secondary observation of a larger pilot study. The Assessing Caregivers for Team Intervention via Video Encounters intervention project enabled caregiver participation in hospice interdisciplinary team meetings. This paper used the team observation scale to assess the impact of caregiver involvement on team structure, process, and outcome. Comparisons were made between traditional team meetings and the team meetings that involved caregivers by using videotapes of both. Analysis found that team meetings with participating caregivers had better team outcomes, with more patient-centered goals, increased discussion of biopsychosocial problems, and the development of interdisciplinary care plans occurring more often than in the traditional hospice team meetings. Findings from this study show benefits for inclusion of caregivers in hospice team meetings. PMID- 20007822 TI - Paul Goldhaber: life and scientific achievements. PMID- 20007821 TI - Electrogenic tuning of the axon initial segment. AB - Action potentials (APs) provide the primary means of rapid information transfer in the nervous system. Where exactly these signals are initiated in neurons has been a basic question in neurobiology and the subject of extensive study. Converging lines of evidence indicate that APs are initiated in a discrete and highly specialized portion of the axon-the axon initial segment (AIS). The authors review key aspects of the organization and function of the AIS and focus on recent work that has provided important insights into its electrical signaling properties. In addition to its main role in AP initiation, the new findings suggest that the AIS is also a site of complex AP modulation by specific types of ion channels localized to this axonal domain. PMID- 20007823 TI - High molecular-weight gelatinase species of human Bruch's membrane: compositional analyses and age-related changes. AB - PURPOSE: The structural and functional demise of aging Bruch's membrane is associated with a reduction in the activity of the matrix metalloproteinase (MMP) degradation system. The gelatinase component of the MMP system consists of MMP2 and MMP9 and two high molecular-weight (HMW1, HMW2) species that are yet to be characterized and whose roles in the aging process are yet to be elucidated. The purpose of this study was to determine the age-related changes in levels of expression and subunit characterization of the HMW gelatinase species of Bruch's membrane. METHODS: Gelatin zymography followed by densitometric scanning was used to quantify the level of the HMW species present. Gel-filtration chromatography allowed the fractionation of the gelatinases according to their molecular weight, and subsequent degradation of the HMW species with a mino-phenyl acetate activation, reduction, and alkylation produced subunit fragments for analysis. RESULTS: Most of the HMW1 and HMW2 pool (80% and 87%, respectively) were tightly bound to the matrix. Aging was associated with significant increases in the levels of HMW1 and HMW2 (P < 0.005 and P < 0.05 respectively). On gel filtration, a single large macromolecular complex (LMMC) was observed containing HMW1, HMW2, MMP9, and some MMP2. Activation-mediated fragmentation of HMW1 and HMW2 showed them to be composed of heteropolymers of MMP2 and MMP9. CONCLUSIONS: The age related increase of HMW1 and HMW2, together with the formation of LMMC, resulted in the sequestration of MMP2 and MMP9, thereby reducing the free pool for activation. This is likely to contribute to reduced matrix degradation and turnover of Bruch's membrane in both normal aging and age-related macular degeneration. PMID- 20007824 TI - Small, hard macular drusen and peripheral drusen: associations with AMD genotypes in the Inter99 Eye Study. AB - PURPOSE: To study associations of small, hard macular drusen and peripheral drusen with genotypes associated with age-related macular degeneration (AMD). METHODS: Digital grayscale fundus photographs recorded in red-free illumination were graded for the presence of drusen in 1107 subjects aged 30 to 66 years. Participants were genotyped for AMD-related polymorphisms in complement factor H (CFH), in LOC387715, and in complement factor B (CFB). RESULTS: The prevalence of 20 or more small, hard macular drusen per eye was 14%, with no association to the investigated polymorphisms. Peripheral drusen were associated with CFHY402H (odds ratio [OR], 4.3; 95% confidence interval [95% CI], 1.4-13, for CC versus TT genotypes) as was macular drusen >63 microm (OR, 1.9; 95% CI, 1.1-3.1, for CC versus TT genotypes). Macular drusen >63 microm were associated with the presence of 20 or more small, hard macular drusen (OR, 1.7; 95% CI, 1.1-2.6) and with peripheral drusen (OR, 2.5; 95% CI,1.2-5.4) CONCLUSIONS: In this study, the presence of 20 or more small, hard macular drusen per eye was not associated with known AMD-related polymorphisms, whereas the study confirmed an association of peripheral drusen with CFHY402H. (ClinicalTrials.gov number, NCT00289237). PMID- 20007825 TI - Inhibition of retinoblastoma in vitro and in vivo with conditionally replicating oncolytic adenovirus H101. AB - PURPOSE: To determine the therapeutic effect of oncolytic adenovirus H101 on retinoblastoma in vitro and in vivo. METHODS: The expression of coxsackievirus adenovirus receptor (CAR) in human retinoblastoma cell line HXO-RB(44) was determined by RT-PCR, Western blot, immunofluorescence, and immunocytochemistry staining. Appropriate multiplicity of infection was determined using flow cytometry in retinoblastoma cells with green fluorescent protein-expressing adenovirus (AdGFP). The viability of HXO-RB(44) cells treated with H101 or AdGFP was measured using a cell counting kit-8-based procedure. Viral proliferation in vitro was measured by end point dilution titration and real-time PCR. Cell cycle and apoptotic activity of HXO-RB(44) were analyzed by flow cytometry. NOD-SCID mice bearing retinoblastoma xenografts were treated with intratumoral injection of H101, AdGFP, or PBS. Tumor volume and survival time were recorded. Immunohistochemistry for adenoviral fiber protein and Western blot for adenoviral Hexon protein of retinoblastoma xenografts were performed to evaluate H101 virus replication in vivo. RESULTS: HXO-RB(44) cells expressed CAR and were sensitive to adenoviral infection. HXO-RB(44) cells treated with H101 had reduced cell viability compared with AdGFP-treated cells (P < 0.01). Abundant replication of H101 in HXO-RB(44) cells resulted in G(2)/M-phase arrest and finally tumor cell lysis, but the apoptosis pathway was not activated. Tumor-bearing mice treated with H101 had reduced tumor burdens and prolonged survival times compared with PBS and AdGFP controls (both P < 0.01). Immunohistochemical and Western blot examination revealed widespread replication of H101 within the tumor. CONCLUSIONS: These results suggest that H101 effectively inhibits the growth of retinoblastoma cells in vitro and in mice and may serve as a novel therapy for retinoblastoma. PMID- 20007826 TI - Paracrine modulation of CXCR4 by IGF-1 and VEGF: implications for choroidal neovascularization. AB - PURPOSE: Modulators of angiogenesis typically work in an orchestrated manner. The authors examined the interaction between insulinlike growth factor (IGF)-1, vascular endothelial growth factor (VEGF), and stromal derived factor (SDF)-1 in vivo and in vitro using angiogenesis models. METHODS: The angiogenic effect of SDF-1, alone or in combination with IGF-1 and VEGF, was assessed in human lung microvascular endothelial cells using capillary tube formation and thymidine incorporation. Immunohistochemical analysis for CD31, SDF-1, and CXCR4 was performed on mouse eyes 2 weeks after the initiation of laser rupture of Bruch's membrane, a choroidal neovascularization (CNV) model. CXCR4 antagonist and CXCR4 blocking antibody were tested on inhibition of CNV lesion size in this model. Real-time PCR was used to determine mRNA levels for SDF-1, VEGF, IGF-1, and their cognate receptors in the retinal pigment epithelium/choroid complex of mice that underwent this CNV model. RESULTS: IGF-1 and VEGF demonstrated an additive effect on SDF-1-induced in vitro angiogenesis. CXCR4 immunoreactivity was present in both normal and laser-injured mice at the laser burn site and at the ganglion cell layer, the anterior portion of the inner nuclear layer, photoreceptors, and choroidal stroma. SDF-1 was observed in identical locations but was not seen in photoreceptors. mRNA levels for SDF-1, VEGF, and IGF-1 and their receptors were increased after laser injury. CXCR4-neutralizing antibody reduced neovascularization when injected subretinally but not intraperitoneally or intravitreally. CONCLUSIONS: The potent proangiogenic factors IGF-1 and VEGF both stimulate SDF-1-induced angiogenesis. Local inhibition of CXCR4 is required for an antiangiogenic effect in CNV lesions. PMID- 20007827 TI - Mechanisms of immune suppression for CD8+ T cells by human corneal endothelial cells via membrane-bound TGFbeta. AB - PURPOSE: To determine whether CD8(+) T cells in vitro are suppressed or converted into regulators (Treg cells) by human corneal endothelial (HCE) cells. METHODS: HCE cell lines were established from primary HCE cells, and allogeneic CD8(+) T cells were isolated from peripheral blood mononuclear cells of healthy donors. T cell activation was assessed for proliferation by [(3)H]-thymidine incorporation. Expression of TGFbeta on HCE cells was evaluated by flow cytometry, RT-PCR, and immunohistochemistry. Anti-TGFbeta-neutralizing antibodies or TGFbeta siRNA was used to abolish the HCE-mediated inhibitory function. CD8(+) T cells exposed to HCE cells were used as Treg cells, to induce Treg cells in vitro by exposure to HCE cells. Expression of CD25 or Foxp3 on Treg cells was evaluated by flow cytometry. RESULTS: Cultured HCE cells produced the membrane-bound active TGFbeta isoform 2. HCE surface TGFbeta2 was necessary, to inhibit CD8(+) T-cell activation via direct cell contact. In addition, although HCE cells were found to constitutively express costimulatory molecules, such as programmed death-ligand 1 (PD-L1) and PD-L2, and secreted thrombosponsin-1, only membrane-bound TGFbeta2 was actually delivered to the CD8(+) T cells. HCE cells were also found to convert some CD8(+) T-cell populations into Treg cells via their membrane-bound TGFbeta. These HCE-induced Treg cells produced soluble forms of TGFbeta1, but not of TGFbeta2, and they also acquired a regulatory phenotype that expressed CD25(high) and Foxp3. CONCLUSIONS: Ocular resident tissue-exposed T cells can be suppressed or induced to become regulators within the human peripheral microenvironment. PMID- 20007828 TI - Suppression of experimental autoimmune uveoretinitis by inducing differentiation of regulatory T cells via activation of aryl hydrocarbon receptor. AB - Purpose. Aryl hydrocarbon receptor (AHR) has been identified as a regulator of CD25(+)CD4(+) regulatory T-cell (T(reg)) and Th17 cell differentiation in mice, and activation of AHR by its ligand 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) induces functional T(reg) cells. In this study, the authors examined whether the AHR-mediated effect of TCDD suppresses mouse experimental autoimmune uveitis (EAU) by inducing T(reg) cell differentiation. Methods. C57BL/6 mice were injected with TCDD 1 day before immunization with human interphotoreceptor retinoid-binding protein peptide 1-20 (hIRBP-p), and the severity of EAU was assessed clinically and histopathologically. Immunologic responses of draining lymph node cells and splenocytes to hIRBP-p and anti-CD3 monoclonal antibody (mAb) were assessed by T-cell proliferation and cytokine production. In addition, differentiation of Foxp3(+) T cells and their immunosuppressive roles in TCDD injected mice were evaluated. Results. TCDD injection increased Foxp3(+) T cells in the lymph nodes and in the spleen. Development of EAU was completely suppressed by TCDD injection, and suppression was abolished by treatment with anti-CD25 mAb before TCDD injection. Both lymphocytes and splenocytes obtained from TCDD-injected mice immunized with hIRBP-p failed to produce IFN-gamma and IL 17 on stimulation with hIRBP-p, and the failure of IL-17 production was observed even when stimulated with anti-CD3 mAb. However, this protocol did not interfere with IL-10 production and T-cell proliferation response when assessed on stimulation with anti-CD3 mAb. Conclusions. Activation of AHR by TCDD markedly suppressed autoimmune uveoretinitis through mechanisms that expand CD25(+)Foxp3(+) T(reg) cells and interfere with the activation of Th1 and Th17 cells. PMID- 20007829 TI - Straylight before and after LASEK in myopia: changes in retinal straylight. AB - PURPOSE: To quantify the changes in retinal straylight that occur after laser assisted subepithelial keratectomy (LASEK). METHODS: This prospective study included 86 eyes of 49 patients who were scheduled for LASEK surgery. Patients were divided into groups based on their preoperative contact lens wear habits: rigid lenses (RCL), soft lenses (SCL), spectacles after a period of contact lenses (SaC), and spectacles only (Specs). Retinal straylight was tested before surgery and 6 months after surgery with the compensation comparison method. Straylight was also compared to a normal reference database. The difference with the average straylight increase with age, called base- and age-corrected (BAC) straylight, was also studied. RESULTS: Before surgery, BAC straylight was found to be strongly elevated, with a value of 0.15 +/- 0.14 log units. After LASEK, this decreased to 0.00 +/- 0.14 log units. The reduction was significant (paired t-test, P << 0.01) and correlated with preoperative BAC straylight levels (r(2) = 0.332; P << 0.01). There was no correlation between the straylight change and the spherical equivalent of the laser refractive correction (r(2) = 0.042; P = 0.059). Preoperative wear of soft contact lenses increased the BAC straylight by approximately 0.06 log units, with respect to the spectacles groups (P < 0.05, unpaired t-test), but after surgery, this difference was no longer found (P > 0.05). CONCLUSIONS: Higher than normal preoperative BAC straylight was found to normalize after LASEK refractive surgery. Wearing soft contact lenses causes an additional increase in preoperative BAC straylight that is eliminated after LASEK. PMID- 20007830 TI - Cone versus rod disease in a mutant Rpgr mouse caused by different genetic backgrounds. AB - PURPOSE: To establish mouse models for RPGR-associated diseases by generating and characterizing an Rpgr mutation (in-frame deletion of exon 4) in two different genetic backgrounds (BL/6 and BALB/c). METHODS: Gene targeting in embryonic stem (ES) cells was performed to introduce a in-frame deletion of exon 4 in the Rpgr gene (Rpgr(DeltaEx4)). Subsequently, the mutation was introduced in two different inbred mouse strains by successive breeding. Mutant and wild-type mice of both strains were characterized by electroretinography (ERG) and histology at five time points (1, 3, 6, 9, and 12 months). RPGR transcript amounts were assessed by quantitative RT-PCR. A variety of photoreceptor proteins, including RPGR-ORF15, RPGRIP, PDE6delta/PrBPdelta, rhodopsin, and cone opsin, were localized on retinal sections by immunohistochemistry. RESULTS: Mislocalization of rhodopsin and cone opsin was an early pathologic event in mutant mice of both lines. In contrast, RPGR-ORF15 as well as RPGRIP1 and PDE6delta/PrBPdelta showed similar localizations in mutant and wild-type animals. Functional and histologic studies revealed a mild rod-dominated phenotype in mutant male mice on the BL/6 background, whereas a cone-dominated phenotype was observed for the same mutation in the BALB/c background. CONCLUSIONS: Both Rpgr mutant mouse lines developed retinal disease with a striking effect of the genetic background. Cone-specific modifiers might influence the retinal phenotype in the BALB/c strain. The two lines provide models to study RPGR function in rods and cones, respectively. PMID- 20007831 TI - Comparison of retinal thickness in normal eyes using Stratus and Spectralis optical coherence tomography. AB - PURPOSE: Spectral-domain optical coherence tomography (SD-OCT) is an advancement over time-domain OCT (TD-OCT) in the imaging of retinal disorders. Retinal thickness measured by SD-OCT differs from that measured by TD-OCT because the delineation of the outer boundary of the retina differs in the two instruments. The present study aims to evaluate this difference by comparing macular thickness, as obtained by Stratus and Spectralis OCT, in subjects without any known retinal disease. METHODS: Thirty-six subjects with no history of retinal disease and with normal vision and normal intraocular pressure were enrolled in the study. Both Stratus and Spectralis OCT scanning were performed by the same operator on all subjects in one eye. Central point thickness (CPT) and retinal thickness in nine ETDRS subfields, including central subfield (CSF), were measured. Student's t-test was used to determine statistical significance. RESULTS: Mean CPT, as measured by the Stratus and Spectralis OCT, was 166.9 +/- 20.9 microm and 225.1 +/- 17.1 microm (P < 0.0001), and mean CSF was 202.3 +/- 19.6 microm and 271.4 +/- 19.6 microm (P < 0.0001), respectively. Although the mean difference in CSF thickness was 69.1 microm, it ranged from 61.9 to 74 microm in the other eight ETDRS subfields. CONCLUSIONS: An increased measurement in retinal thickness of approximately 65 to 70 microm, as measured by Spectralis OCT compared with Stratus OCT, is consistent with the extent of axial retinal thickness measured by the two instruments. This increased measurement corresponds to the inclusion of the outer segment-RPE-Bruch's membrane complex by Spectralis OCT, which is relevant to studies using the newer SD-OCT for assessment of retinal thickness. PMID- 20007832 TI - Post-illumination pupil response in subjects without ocular disease. AB - PURPOSE: A sustained pupilloconstriction is often observed after the cessation of a bright visual stimulus. This post-illumination pupil response (PIPR) is produced by the intrinsically photosensitive retinal ganglion cells (ipRGCs). The present study was designed to examine the characteristics of the PIPR in a normal population without ocular disease. METHODS: Thirty-seven subjects (mean age, 48.6 years) were tested by presenting a 60 degrees, 10-second light stimulus (13 log quanta/cm(2)/s retinal irradiance) and recording pupillary responses for 50 seconds after light cessation. The light stimuli (470 [blue] and 623 [red] nm) were presented by an optical system to one eye after dilation, while the consensual pupil response of the fellow, undilated eye was recorded by infrared pupillometry. RESULTS: A positive PIPR was seen in all subjects tested. The population average of the PIPR for 470-nm light was 1.5 mm (SEM 0.10, P < 0.05) and the net PIPR (blue PIPR minus red PIPR) was 1.4 mm (SEM 0.09, P < 0.0001). The net PIPR correlated positively with baseline pupil diameter (P < 0.05), but not significantly with age, race, or sex (P > 0.05) in the test population. CONCLUSIONS: All normal subjects displayed a significant PIPR for a 10-second, 470-nm light stimulus, but not a 623-nm stimulus, which is consistent with the proposed melanopsin-mediated response. In most normal individuals, the amplitude of the PIPR was substantial. This test has the potential to be used as a tool in evaluating subjects with inner retinal dysfunction or melanopsin-related disorders. PMID- 20007833 TI - Insulin-like growth factor-1 and cardiotrophin 1 increase strength and mass of extraocular muscle in juvenile chicken. AB - PURPOSE: Insulin-like growth factor 1 (IGF1) and cardiotrophin 1 (CT1) are known to increase the strength of extraocular muscles in adult and embryonic animals, but no information is available for the early postnatal period, when strabismus treatment in humans is most urgent. Here the authors sought to determine whether these trophic factors strengthen juvenile maturing extraocular muscles and gain insight into mechanisms of force increase. METHODS: After two injections of IGF1, CT1, or both with different dosages in posthatch chickens, the authors quantified five parameters of the superior oblique extraocular muscle at 2 weeks of age: contractile force, muscle mass, total myofiber area, myofiber diameter, and number of proliferating satellite cells labeled by bromodeoxyuridine. RESULTS: Treatment with IGF1, CT1, and combination of IGF1 and CT1 significantly increased contractile force by 14% to 22%. CT1 and combination treatment significantly increased muscle mass by 10% to 24%. IGF1/CT1 combination treatment did not have additive effects on strengthening muscles, compared with single-drug treatments. Myofiber area increased significantly with IGF1 and CT1 treatment in proximal, but not distal, parts of the muscle and this was due to increased fiber numbers or length (IGF1) or increased diameters of global layer myofibers (CT1). Trophic factors increased the number of proliferating (bromodeoxyuridine-labeled) satellite cells in proximal and middle segments of muscles. CONCLUSIONS: Exogenous IGF1 and CT1 strengthen extraocular muscles during maturation. They predominantly remodel the proximal segment of juvenile extraocular muscles. This information about muscle plasticity may aid the design of pharmacologic treatment of strabismus in children during the "critical period" of oculomotor maturation. PMID- 20007834 TI - Proliferative response of microglia and macrophages in the adult mouse eye after optic nerve lesion. AB - PURPOSE: The purpose of this in vivo study was to evaluate the proliferative response of immunologic cells during the acute phase after optic nerve (ON) lesion in the neural retina and the ciliary body (CB) in the adult mouse. METHODS: The number of cells obtained 5 to 10 days after ON crush was compared with that counted after intraorbital ON transection. In addition, after ON crush, the time course of in situ proliferating Ki67(+) microglia and macrophages was analyzed from 6 hours up to 10 days. RESULTS: The number of BrdU(+)F4/80(+) retinal microglia and ciliary macrophages increased over time, reaching the peak number 10 days after ON lesion. In the retina, both ON lesion types resulted in a similar number of BrdU(+)F4/80(+) microglia. Approximately 85% of all BrdU(+) cells were identified as F4/80(+) microglia. However, this cell population represented only 30% of all F4/80(+) microglia. The peak of microglial in situ proliferation was found 2 days after ON crush. In the CB, both ON lesion types induced a significant increase in the number of BrdU(+)F4/80(+) macrophages. Of interest, the number of cells after ON transection further increased over time, whereas those after ON crush did not. CONCLUSIONS: ON lesion significantly increased proliferation of F4/80(+) immunologic cells in both the retina and CB. Although no significant differences in cellular response were observed in the retina between both lesion types, ON transection had a more pronounced effect on ciliary macrophages than did ON crush. Therefore, both regions seem not to act in concert during the acute phase after ON lesion. PMID- 20007835 TI - Structural effects of fibulin 5 missense mutations associated with age-related macular degeneration and cutis laxa. AB - PURPOSE: AMD has a complex etiology with environmental and genetic risk factors. Ten fibulin 5 sequence variants have been associated with AMD and two other fibulin 5 mutations cause autosomal-recessive cutis laxa. Fibulin 5 is a 52-kDa calcium-binding epidermal growth factor (cbEGF)-rich extracellular matrix protein that is essential for the formation of elastic tissues. Biophysical techniques were used to detect structural changes in the fibulin 5 mutants and to determine whether changes are predictive of pathogenicity. METHODS: Native PAGE, nonreduced SDS-PAGE, size-exclusion column multiangle laser light scattering, sedimentation velocity, and circular dichroism (CD) were used to investigate the mobility, hydrodynamic radii, folding, and oligomeric states of the fibulin 5 mutants in the absence and presence of Ca(2+). RESULTS: CD showed that all mutants are folded, although perturbations to secondary structure contents were detected. Both cutis laxa mutants increased dimerization. Most other mutants slightly increased self-association in the absence of Ca(2+) but this was also demonstrated by G202R, a polymorphism detected in a control individual. The AMD associated mutant G412E showed lower-than-expected mobility during native-PAGE, the largest hydrodynamic radius for the monomer form and the highest levels of aggregation in both the absence and presence of Ca(2+). CONCLUSIONS: The results identified structural differences for the disease-causing cutis laxa mutants and for one AMD variant (G412E), suggesting that this may also be pathogenic. Although the other AMD-associated mutants showed no gross structural differences, they cannot be excluded as pathogenic by differences outside the scope of this study-for example, disruption of heterointeractions. PMID- 20007836 TI - Alterations in the intraocular cytokine milieu after intravitreal bevacizumab. AB - PURPOSE: Several complications after intravitreal bevacizumab (IVB) treatment have been described including tears of the retinal pigment epithelium and tractional retinal detachment. The etiology of these complications remains unclear. The purpose of this study was to characterize changes in the intraocular levels of inflammatory cytokines after IVB as a possible explanation for these complications. METHODS: Twenty-nine patients with proliferative diabetic retinopathy (PDR) undergoing pars plana vitrectomy (PPV) for vitreous hemorrhage (VH) with IVB pretreatment were prospectively enrolled. Aqueous humor samples were taken at the time of IVB pretreatment and approximately 1 week later at the time of PPV. Multiplex cytokine arrays were used to assay 20 different cytokines. Multivariate general linear regression was performed to determine differences in cytokine levels between the two study visits. Proportional hazards regression was performed to determine the relationship between cytokine levels at PPV and postoperative outcomes. RESULTS: After treatment with IVB, vascular endothelial growth factor (VEGF) concentrations in the aqueous humor decreased (P = 0.0003), whereas the concentrations of IL-8 and transforming growth factor (TGF)-beta(2) increased after IVB (P < 0.03). The level of IL-8 at the time of PPV was associated with the occurrence of recurrent VH after surgery (hazard ratio, 1.32; P = 0.02). CONCLUSIONS: Alterations in the intraocular inflammatory cytokine milieu occur after IVB injection, possibly as a compensatory mechanism in response to VEGF inhibition. The increased concentrations of inflammatory cytokines after IVB may be clinically significant and may be responsible for some of the complications after IVB. PMID- 20007837 TI - Epifluorescence intravital microscopy of murine corneal dendritic cells. AB - Purpose. Dendritic cells (DCs) are antigen-presenting cells vital for initiating immune responses. In this study the authors examined the in vivo migratory capability of resident corneal DCs to various stimuli. Methods. The authors used mice expressing enhanced yellow fluorescent protein (eYFP) under control of the CD11c promoter to visualize corneal DCs. To assess the distribution and mobility of DCs, normal corneas were imaged in vivo and ex vivo with fluorescence microscopy. Intravital microscopy was used to examine the responses of resident central and peripheral corneal DCs to silver nitrate injury, lipopolysaccharide, microspheres, and tumor necrosis factor (TNF-alpha). In some experiments, TNF alpha injection was used to first induce centripetal migration of DCs to the central cornea, which was subsequently reinjected with microspheres. Results. In normal corneas, DCs were sparsely distributed centrally and were denser in the periphery, with epithelial-level DCs extending into the epithelium. Videomicroscopy showed that though cell processes were in continuous movement, cells generally did not migrate. Within the first 6 hours after stimulation, neither central nor peripheral corneal DCs exhibited significant lateral migration, but central corneal DCs assumed extreme morphologic changes. An increased number of DCs in the TNF-alpha-stimulated central cornea were responsive to subsequent microsphere injection by adopting a migratory behavior, but not with increased speed. Conclusions. In vivo imaging reveals minimal lateral migration of corneal DCs after various stimuli. In contrast, DCs within the central cornea after initial TNF-alpha injection are more likely to respond to a secondary insult with lateral migration. PMID- 20007838 TI - The crucial role of ATGL for energy supply of muscles. PMID- 20007839 TI - The role of CD4+CD25+ regulatory T cells in macrophage-derived foam-cell formation. AB - Cluster of differentiation (CD)4+CD25+ regulatory T cells (Tregs) exert a suppressive activity on atherosclerosis, but the underlying mechanism remains unclear. Here, we investigated whether and how Tregs affect macrophages foam-cell formation. Tregs were isolated by magnetic cell sorting-column and analyzed by flow cytometry. Macrophages were cultured with or without Tregs in the presence of oxidized LDL (oxLDL) for 48 h to transform foam cells. After co-culture with Tregs, macrophages showed a decrease in lipid accumulation, which was accompanied by a significantly downregulated expression of CD36 and SRA but no obvious difference in ABCA1 expression. Tregs can inhibit the proinflammatory properties of macrophages and steer macrophage differentiation toward an anti-inflammatory cytokine producing phenotype. Mechanistic studies reveal that both cell-to-cell contact and soluble factors are required for Treg-mediated suppression on macrophage foam-cell formation. Cytokines, interleukin-10 (IL-10), and transforming growth factor-beta (TGF-beta) are the key factors for these suppressive functions. PMID- 20007840 TI - Usefulness of peptide nucleic acid (PNA)-clamp smart amplification process version 2 (SmartAmp2) for clinical diagnosis of KRAS codon 12 mutations in lung adenocarcinoma: comparison of PNA-clamp SmartAmp2 and PCR-related methods. AB - KRAS is an oncogene that can be activated by mutations. Patients with non-small cell lung cancer who have KRAS mutations do not respond to tyrosine kinase inhibitors; therefore, accurate detection of KRAS mutations is important for deciding therapeutic strategies. Although sequencing-related techniques have been frequently used, they are usually too complex, have low sensitivity, and are time consuming for routine screening in clinical situations. We evaluated peptide nucleic acid (PNA)-clamp smart amplification process version 2 (SmartAmp2) as a detection method for KRAS codon 12 mutations in patient specimens compared with traditional sequencing and polymerase chain reaction-related methods. Among 172 lung adenocarcinoma samples, direct sequencing, enzyme-enriched sequencing, and PNA-enriched sequencing showed that 16 (9.3%), 26 (15.7%), and 28 (16.3%) tumors, respectively, contained KRAS mutations in codon 12. Using PNA-clamp SmartAmp2, we could identify 31 (18.0%) tumors that had KRAS mutations in codon 12 within 60 minutes, three of which were undetected by polymerase chain reaction-related methods. On the other hand, we examined 30 nonmalignant peripheral lung tissue specimens and found no mutations in any of the samples using PNA-clamp SmartAmp2. In this study, we confirmed that PNA-clamp SmartAmp2 has high sensitivity and accuracy and is suitable for the clinical diagnosis of KRAS codon 12 mutations. PMID- 20007841 TI - KRAS genotyping of paraffin-embedded colorectal cancer tissue in routine diagnostics: comparison of methods and impact of histology. AB - KRAS mutation testing before anti-epidermal growth factor receptor therapy of metastatic colorectal cancer has become mandatory in Europe. However, considerable uncertainty exists as to which methods for detection can be applied in a reproducible and economically sound manner in the routine diagnostic setting. To answer this question, we examined 263 consecutive routine paraffin slide specimens. Genomic DNA was extracted from microdissected tumor tissue. The DNA was analyzed prospectively by Sanger sequencing and array analysis as well as retrospectively by melting curve analysis and pyrosequencing; the results were correlated to tissue characteristics. The methods were then compared regarding the reported results, costs, and working times. Approximately 40% of specimens contained KRAS mutations, and the different methods reported concordant results (kappa values >0.9). Specimens harboring fewer than 10% tumor cells showed lower mutation rates regardless of the method used, and histoanatomical variables had no influence on the frequency of the mutations. Costs per assay were higher for array analysis and melting curve analysis when compared with the direct sequencing methods. However, for sequencing methods equipment costs were much higher. In conclusion, Sanger sequencing, array analysis, melting curve analysis, and pyrosequencing were equally effective for routine diagnostic KRAS mutation analysis; however, interpretation of mutation results in conjunction with histomorphologic tissue review and on slide tumor tissue dissection is required for accurate diagnosis. PMID- 20007842 TI - Molecular diagnostics for invasive fungal infections: a call for refinement and implementation. PMID- 20007843 TI - Combined iPLEX and TaqMan assays to screen for 45 common mutations in Lynch syndrome and FAP patients. AB - Mutations of genes associated with the mismatch repair mechanism and mutations of the APC gene are the most frequent causes of hereditary colorectal cancer. An iPLEX test combined with TaqMan genotyping assays was therefore developed to identify common recurrent mutations of those genes in the Polish population. We analyzed 349 DNA samples from 95 positive controls previously identified by sequencing and 254 unexamined individuals. The iPLEX test included two plexes, which comprised seven mutations of the APC gene and 29 mutations of three of the mismatch repair genes. TaqMan assays were designed for nine mutations not covered by the iPLEX assays: one mutation in the APC gene and eight mutations in the mismatch repair genes. Results were then verified independently by sequencing. Our combination method allowed detection of all recurrent mutations occurring in group of patients, followed by full analysis by DNA sequencing. With the exception of one false positive in the iPLEX test in the positive control group that could be assigned to contamination from neighboring wells rather than a detection error, given sufficient DNA concentration and quality, the designed iPLEX/TaqMan test had an accuracy of 100% for the designed assays. These results suggest that the combined iPLEX/TaqMan test is an outstanding tool for identification of recurrent mutations among hereditary colorectal cancer patients. PMID- 20007844 TI - Modeling complex workflow in molecular diagnostics: design specifications of laboratory software for support of personalized medicine. AB - One of the hurdles to achieving personalized medicine has been implementing the laboratory processes for performing and reporting complex molecular tests. The rapidly changing test rosters and complex analysis platforms in molecular diagnostics have meant that many clinical laboratories still use labor-intensive manual processing and testing without the level of automation seen in high-volume chemistry and hematology testing. We provide here a discussion of design requirements and the results of implementation of a suite of lab management tools that incorporate the many elements required for use of molecular diagnostics in personalized medicine, particularly in cancer. These applications provide the functionality required for sample accessioning and tracking, material generation, and testing that are particular to the evolving needs of individualized molecular diagnostics. On implementation, the applications described here resulted in improvements in the turn-around time for reporting of more complex molecular test sets, and significant changes in the workflow. Therefore, careful mapping of workflow can permit design of software applications that simplify even the complex demands of specialized molecular testing. By incorporating design features for order review, software tools can permit a more personalized approach to sample handling and test selection without compromising efficiency. PMID- 20007845 TI - KRAS mutation: comparison of testing methods and tissue sampling techniques in colon cancer. AB - Treatment of colon carcinoma with the anti-epidermal growth factor receptor antibody Cetuximab is reported to be ineffective in KRAS-mutant tumors. Mutation testing techniques have therefore become an urgent concern. We have compared three methods for detecting KRAS mutations in 59 cases of colon carcinoma: 1) high resolution melting, 2) the amplification refractory mutation system using a bifunctional self-probing primer (ARMS/Scorpion, ARMS/S), and 3) direct sequencing. We also evaluated the effects of the methods of sectioning and coring of paraffin blocks to obtain tumor DNA on assay sensitivity and specificity. The most sensitive and specific combination of block sampling and mutational analysis was ARMS/S performed on DNA derived from 1-mm paraffin cores. This combination of tissue sampling and testing method detected KRAS mutations in 46% of colon tumors. Four samples were positive by ARMS/S, but initially negative by direct sequencing. Cloned DNA samples were retested by direct sequencing, and in all four cases KRAS mutations were identified in the DNA. In six cases, high resolution melting abnormalities could not be confirmed as specific mutations either by ARMS/S or direct sequencing. We conclude that coring of the paraffin blocks and testing by ARMS/S is a sensitive, specific, and efficient method for KRAS testing. PMID- 20007846 TI - Identification of a gene for renal-hepatic-pancreatic dysplasia by microarray based homozygosity mapping. AB - We have investigated a family where two siblings had a developmental disorder associated with polycystic dysplastic kidney disease that was incompatible with postnatal survival. Additional features observed were ductal plate malformation in the liver, dysplasia of the pancreas, and (in one individual) complete situs inversus and polymicrogyria of the cingulate gyri. The autopsy findings were compatible with renal-hepatic-pancreatic dysplasia, a condition with unknown genetic cause at the time of autopsy but with similarities to the Meckel Gruber/Joubert group of recessive ciliopathies. Consanguinity between the parents made it likely that the mutated gene (with known or potential function in cilia) was located within a rather large region of homozygosity in the affected individuals (identical by descent). Using genetic markers (50K single nucleotide polymorphism microarrays), we found a single large homozygous region of 21.16 Mb containing approximately 200 genes on the long arm of chromosome 3. This region contained two known ciliopathy genes: NPHP3 (adolescent nephronophthisis) and IQCB1 (NPHP5), which is associated with Senior-Loken syndrome. In NPHP3, homozygosity for a deletion of the conserved splice acceptor dinucleotide (AG) preceding exon 20 was found. Our finding confirms the recent report that NPHP3 null mutations cause renal-hepatic-pancreatic dysplasia. Also, our case illustrates that genes for rare and genetically heterogeneous recessive conditions may be identified by homozygosity mapping using single nucleotide polymorphism arrays in the routine clinical setting. PMID- 20007848 TI - High-throughput identification and quantification of Candida species using high resolution derivative melt analysis of panfungal amplicons. AB - Fungal infections pose unique challenges to molecular diagnostics; fungal molecular diagnostics consequently lags behind bacterial and viral counterparts. Nevertheless, fungal infections are often life-threatening, and early detection and identification of species is crucial to successful intervention. A high throughput PCR-based method is needed that is independent of culture, is sensitive to the level of one fungal cell per milliliter of blood or other tissue types, and is capable of detecting species and resistance mutations. We introduce the use of high resolution melt analysis, in combination with more sensitive, inclusive, and appropriately positioned panfungal primers, to address these needs. PCR-based amplification of the variable internal transcribed regions of the rDNA genes generates an amplicon whose sequence melts with a shape that is characteristic and therefore diagnostic of the species. Simple analysis of the differences between test and reference melt curves generates a single number that calls the species. Early indications suggest that high resolution melt analysis can distinguish all eight major species of Candida of clinical significance without interference from excess human DNA. Candida species, including mixed and novel species, can be identified directly in vaginal samples. This tool can potentially detect, count, and identify fungi in hundreds of samples per day without further manipulation, costs, or delays, offering a major step forward in fungal molecular diagnostics. PMID- 20007849 TI - 5-HT(1A), SST(1), and SST(2) receptors mediate inhibitory postsynaptic potentials in the submucous plexus of the guinea pig ileum. AB - Vasoactive intestinal peptide (VIP) immunoreactive neurons are important secretomotor neurons in the submucous plexus. They are the only submucosal neurons to receive inhibitory inputs and exhibit both noradrenergic and nonadrenergic inhibitory synaptic potentials (IPSPs). The former are mediated by alpha(2)-adrenoceptors, but the receptors mediating the latter have not been identified. We used standard intracellular recording, RT-PCR, and confocal microscopy to test whether 5-HT(1A), SST(1), and/or SST(2) receptors mediate nonadrenergic IPSPs in VIP submucosal neurons in guinea pig ileum in vitro. The specific 5-HT(1A) receptor antagonist WAY 100135 (1 microM) reduced the amplitude of IPSPs, an effect that persisted in the presence of the alpha(2)-adrenoceptor antagonist idazoxan (2 microM), suggesting that 5-HT might mediate a component of the IPSPs. Confocal microscopy revealed that there were many 5-HT-immunoreactive varicosities in close contact with VIP neurons. The specific SSTR(2) antagonist CYN 154806 (100 nM) and a specific SSTR(1) antagonist SRA 880 (3 microM) each reduced the amplitude of nonadrenergic IPSPs and hyperpolarizations evoked by somatostatin. In contrast with the other antagonists, CYN 154806 also reduced the durations of nonadrenergic IPSPs. Effects of WAY 100135 and CYN 154806 were additive. RT-PCR revealed gene transcripts for 5-HT(1A), SST(1), and SST(2) receptors in stripped submucous plexus preparations consistent with the pharmacological data. Although the involvement of other neurotransmitters or receptors cannot be excluded, we conclude that 5-HT(1A), SST(1), and SST(2) receptors mediate nonadrenergic IPSPs in the noncholinergic (VIP) secretomotor neurons. This study thus provides the tools to identify functions of enteric neural pathways that inhibit secretomotor reflexes. PMID- 20007850 TI - BMP2 promotes differentiation of nitrergic and catecholaminergic enteric neurons through a Smad1-dependent pathway. AB - The bone morphogenetic protein (BMP) family is a class of transforming growth factor (TGF-beta) superfamily molecules that have been implicated in neuronal differentiation. We studied the effects of BMP2 and glial cell line-derived neurotrophic factor (GDNF) on inducing differentiation of enteric neurons and the signal transduction pathways involved. Studies were performed using a novel murine fetal enteric neuronal cell line (IM-FEN) and primary enteric neurons. Enteric neurons were cultured in the presence of vehicle, GDNF (100 ng/ml), BMP2 (10 ng/ml), or both (GDNF + BMP2), and differentiation was assessed by neurite length, markers of neuronal differentiation (neurofilament medium polypeptide and beta-III-tubulin), and neurotransmitter expression [neuropeptide Y (NPY), neuronal nitric oxide synthase (nNOS), tyrosine hydroxylase (TH), choline acetyltransferase (ChAT) and Substance P]. BMP2 increased the differentiation of enteric neurons compared with vehicle and GDNF-treated neurons (P < 0.001). BMP2 increased the expression of the mature neuronal markers (P < 0.05). BMP2 promoted differentiation of NPY-, nNOS-, and TH-expressing neurons (P < 0.001) but had no effect on the expression of cholinergic neurons (ChAT, Substance P). Neurons cultured in the presence of BMP2 have higher numbers of TH-expressing neurons after exposure to 1-methyl 4-phenylpyridinium (MPP(+)) compared with those cultured with MPP(+) alone (P < 0.01). The Smad signal transduction pathway has been implicated in TGF-beta signaling. BMP2 induced phosphorylation of Smad1, and the effects of BMP2 on differentiation of enteric neurons were significantly reduced in the presence of Smad1 siRNA, implicating the role of Smad1 in BMP2 induced differentiation. The effects of BMP2 on catecholaminergic neurons may have therapeutic implications in gastrointestinal motility disturbances. PMID- 20007852 TI - Force depression in single myofibrils. AB - Force depression after active shortening has been observed in different muscle preparations. It has been assumed that force depression is caused by the development of sarcomere length nonuniformities after shortening. However, this hypothesis has never been investigated in a preparation where individual sarcomere lengths could be directly measured. Here, we investigated force depression in single myofibrils (n = 11) and tracked simultaneously the changes in individual sarcomere lengths (n = 60) before, during, and after shortening and after a purely isometric contraction performed at the final length. Shortening produced force depression in all myofibrils (mean +/- SE; 30.9 +/- 3.9%). During shortening, all sarcomeres shortened, but not by the same amount. Sarcomere lengths were nonuniform, with the same mean SD before (0.11 +/- 0.06 microm) and after shortening (0.11 +/- 0.06 microm) and after a purely isometric contraction at the final length (0.10 +/- 0.05 microm). Furthermore, greater shortening magnitudes were found for sarcomeres that were long in the initial isometric configuration. Nonuniformities of half-sarcomere lengths were also the same before (SD = 0.13 microm) and after (SD = 0.14 microm) shortening. We conclude from these results that the development of sarcomere (or half-sarcomere) length nonuniformities does not play a major role in force depression. Rather, force depression seems an intrinsic property of individual (half-) sarcomeres and muscle contraction. PMID- 20007851 TI - Involvement of adiponectin-SIRT1-AMPK signaling in the protective action of rosiglitazone against alcoholic fatty liver in mice. AB - The development of alcoholic fatty liver is associated with reduced adipocyte derived adiponectin levels, decreased hepatic adiponectin receptors, and deranged hepatic adiponectin signaling in animals. Peroxisomal proliferator-activated receptor-gamma (PPAR-gamma) plays a key role in the regulation of adiponectin in adipose tissue. The aim of the present study was to test the ability of rosiglitazone, a known PPAR-gamma agonist, to reverse the inhibitory effects of ethanol on adiponectin expression and its hepatic signaling, and to attenuate alcoholic liver steatosis in mice. Mice were fed modified Lieber-DeCarli ethanol containing liquid diets for 4 wk or pair-fed control diets. Four groups of mice were given a dose of either 3 or 10 mg.kg body wt(-1).day(-1) of rosiglitazone with or without ethanol in their diets for the last 2 wk of the feeding study. Coadministration of rosiglitazone and ethanol increased the expression and circulating levels of adiponectin and enhanced the expression of hepatic adiponectin receptors (AdipoRs) in mice. These increases correlated closely with the activation of a hepatic sirtuin 1 (SIRT1)-AMP-activated kinase (AMPK) signaling system. In concordance with stimulated SIRT1-AMPK signaling, rosiglitazone administration enhanced expression of fatty acid oxidation enzymes, normalized lipin 1 expression, and blocked elevated expression of genes encoding lipogenic enzymes which, in turn, led to increased fatty acid oxidation, reduced lipogenesis, and alleviation of steatosis in the livers of ethanol-fed mice. Enhanced hepatic adiponectin-SIRT1-AMPK signaling contributes, at least in part, to the protective action of rosiglitazone against alcoholic fatty liver in mice. PMID- 20007853 TI - Quantitative analysis of exercise-induced enhancement of early- and late-systolic retrograde coronary blood flow. AB - Coronary blood flow (CBF) is reduced and transiently reversed during systole via cardiac contraction. Cardiac contractility, coronary tone, and arterial pressure each influence systolic CBF (CBF(SYS)), particularly by modulating the retrograde component of CBF(SYS). The effect of concurrent changes in these factors on CBF(SYS) during dynamic exercise has not been examined. Using chronically instrumented swine, we hypothesized that dynamic exercise enhances retrograde CBF(SYS). Phasic CBF was examined at rest and during treadmill exercise [2-5 miles/h (mph)]. Absolute values of mean CBF over the cardiac cycle (CBF(CYCLE)) as well as mean CBF in diastole (CBF(DIAS)) and mean CBF(SYS) were increased by exercise, while relative CBF(DIAS) and CBF(SYS) expressed as percentage of mean CBF(CYCLE) were principally unchanged. Early retrograde CBF(SYS) was present at rest and increased in magnitude (-33 +/- 4 ml/min) and as a percent of CBF(CYCLE) (-0.6 +/- 0.1%) at 5 mph. This reversal was transient, comprising 3.7 +/- 0.3% of cardiac cycle duration at 5 mph. Our results also reveal that moderately intense exercise (>3 mph) induced a second CBF reversal in late systole before aortic valve closure. At 5 mph, late retrograde CBF(SYS) amounted to -53 +/- 11 ml/min ( 3.1 +/- 0.7% of CBF(CYCLE)) while occupying 11.1 +/- 0.3% of cardiac cycle duration. Wave-intensity analysis revealed that the second flow reversal coincided with an enhanced aortic forward-going decompression wave (vs. rest). Therefore, our data demonstrate a predictable increase in early-systolic CBF reversal during exercise and additionally that exercise induces a late-systolic CBF reversal related to the hemodynamic effects of left ventricular relaxation that is not predictable using current models of phasic CBF. PMID- 20007854 TI - Resting and exercise-induced IL-6 levels in children with Type 1 diabetes reflect hyperglycemic profiles during the previous 3 days. AB - Poor glycemic control in Type 1 diabetes (T1DM) causes long-term cardiovascular complications, at least in part via chronic, low-grade inflammation associated with recurrent hyperglycemia. While physical activity can reduce both inflammation and cardiovascular risks, the underlying molecular mechanisms remain unclear. This is particularly important for T1DM children, for whom the prevention of long-term cardiovascular complications must include optimization of exercise-related anti-inflammatory strategies. We therefore studied the effect of prior hyperglycemia on resting and exercise-induced inflammatory status (plasma IL-6) in T1DM children. Glycemia was continuously recorded with a continuous glucose monitoring system (CGMS) system for 63 h preceding a 30-min intermittent cycling exercise protocol at approximately 80% peak rate of oxygen uptake (VO2max). Euglycemia (4.4-6.1 mM) was maintained for 90 min before, during, and 30 min after exercise. IL-6 plasma concentration (pg/ml) was measured at baseline, at end exercise, and 30 min postexercise. Subjects were then divided into quartiles based on average glycemia during the CGMS recording. IL-6 levels (pg/ml) were lowest in the quartile with lowest average 3-day glycemia and increased proportionally to greater hyperglycemic exposure; this was observed at baseline (0.86 +/- 0.10, 1.06 +/- 0.16, 1.14 +/- 0.14, 1.20 +/- 0.16), absolute IL-6 change (Delta) at end exercise (0.20 +/- 0.16, 0.32 +/- 0.10, 0.48 +/- 0.09, 0.62 +/- 0.13), and Delta at 30 min postexercise (0.49 +/- 0.13, 0.71 +/- 0.16, 0.89 +/- 0.14, 1.38 +/- 0.33). Therefore, poorly controlled glycemic profile, even in the 63 h preceding an exercise challenge, can alter inflammatory adaptation in T1DM children. Our data underscore the necessity to fully understand all molecular aspects of physical activity to provide the scientific rationale for exercise regimens that will be able to maximize health benefits for T1DM children. PMID- 20007855 TI - Local ascorbate administration inhibits the adrenergic vasoconstrictor response to local cooling in the human skin. AB - Local cooling (LC) of nonglabrous skin causes vasoconstriction via the adrenergic and removal of nitric oxide (NO) systems. Since cooling increases reactive oxygen species in smooth muscle cells and induces increased sensitivity of alpha adrenergic receptors, antioxidant supplementation may attenuate the vasoconstrictor response to skin LC via adrenergic and/or NO systems. To test this hypothesis, we examined the effects of acute L-ascorbate (Asc, 10 mM) supplementation in human skin on the vasoconstrictor responses to LC in skin with and without NO synthase (NOS) inhibition or adrenergic receptor blockade. In a three-part study, forearm sites were instrumented with microdialysis fibers, local coolers, and laser-Doppler flow (LDF) probes in healthy volunteers. Sites were cooled from 34 to 24 degrees C at -1 degrees C/min and maintained at 24 degrees C for 20 min (parts 1 and 2) or 30 min (part 3). During the last 10 min of LC in parts 1 and 2, whole body cooling was performed to increase sympathetic vasoconstrictor activity. Cutaneous vascular conductance (CVC) was calculated as the ratio of LDF to blood pressure and expressed relative to the baseline value before cooling. Treatments in each part were as follows: part 1) untreated, Asc; part 2) N(G)-nitro-L-arginine methyl ester (L-NAME) to inhibit NOS, combined L NAME + Asc; part 3) yohimbine (YOH) + propranolol (PRO) to antagonize alpha- and beta-adrenergic receptors and combined YOH + PRO + Asc. CVC reduction during LC was smaller (P < 0.001) at Asc sites (-31 +/- 4%) than at untreated sites (-56 +/ 5%). LC-induced reduction in CVC was smaller (P < 0.05) at L-NAME + Asc sites ( 23 +/- 8%) than at L-NAME sites (-43 +/- 7%). LC-induced reduction in CVC did not differ between at PRO + YOH sites (-56 +/- 3%) and at PRO + YOH + Asc sites (-50 +/- 3%). These findings suggest that antioxidant supplementation inhibits the vasoconstrictor response to direct cooling through an adrenoceptor-dependent mechanism in human skin. PMID- 20007857 TI - Muscle blood flow responses to dynamic exercise in young obese humans. AB - Exercise is a common nonpharmacological way to combat obesity; however, no studies have systematically tested whether obese humans exhibit reduced skeletal muscle blood flow during dynamic exercise. We hypothesized that exercise-induced blood flow to skeletal muscle would be lower in young healthy obese subjects (body mass index of >30 kg/m(2)) compared with lean subjects (body mass index of <25 kg/m(2)). We measured blood flow (Doppler Ultrasound of the brachial and femoral arteries), blood pressure (auscultation, Finapress), and heart rate (ECG) during rest and two forms of single-limb, steady-state dynamic exercise: forearm exercise (20 contractions/min at 4, 8, and 12 kg) and leg exercise (40 kicks/min at 7 and 14 W). Forearm exercise increased forearm blood flow (FBF) similarly in both groups (P > 0.05; obese subjects n = 9, lean subjects n = 9). When FBF was normalized for perfusion pressure, forearm vascular conductance was not different between groups at increasing workloads (P > 0.05). Leg exercise increased leg blood flow (LBF) similarly in both groups (P > 0.05; obese subjects n = 10, lean subjects n = 12). When LBF was normalized for perfusion pressure, leg vascular conductance was not different between groups at increasing workloads (P > 0.05). These results were confirmed when relative blood flow was expressed at average relative workloads. In conclusion, our results show that obese subjects exhibited preserved FBF and LBF during dynamic exercise. PMID- 20007856 TI - Sensitivity of 24-h EMG duration and intensity in the human vastus lateralis muscle to threshold changes. AB - Few studies have quantified lower limb muscle activity over 24 h using electromyographic signals (EMG). None have described the changes in EMG duration and intensity when data are analyzed with different thresholds. Continuous bilateral EMG recordings were made from vastus lateralis (VL) in 10 subjects (20 48 yr) for 24 h. Before and after this recording, voluntary quadriceps forces and VL EMG at 25%, 50%, 75%, and 100% of the maximal voluntary contraction (MVC), percentage voluntary activation (twitch interpolation), and compound action potentials (M-waves) were recorded. Offline, the 24-h EMG integrals (IEMG, 10-ms time constant) were normalized to the MVC IEMG. Total EMG duration and mean IEMG ranged from 1-3 h and 3.2-12.1% MVC, respectively, when the data were analyzed using the baseline (+3 SD) as threshold. When analysis was done with progressively higher thresholds, from baseline up to 4% MVC, the total EMG duration declined curvilinearly. In some cases the decline in duration was 50-60% for a 1% MVC threshold increment. The mean 24-h IEMG increased by 1.5-2% MVC for each 1% MVC threshold increment. Hence, a small change in the analysis threshold may result in large changes in 24-h EMG duration but moderate changes in mean IEMG. Our findings suggest that VL was active for a short amount of time and at low intensities over 24 h. PMID- 20007858 TI - Protein-based multiplex assays: mock presubmissions to the US Food and Drug Administration. AB - As a part of ongoing efforts of the NCI-FDA Interagency Oncology Task Force subcommittee on molecular diagnostics, members of the Clinical Proteomic Technology Assessment for Cancer program of the National Cancer Institute have submitted 2 protein-based multiplex assay descriptions to the Office of In Vitro Diagnostic Device Evaluation and Safety, US Food and Drug Administration. The objective was to evaluate the analytical measurement criteria and studies needed to validate protein-based multiplex assays. Each submission described a different protein-based platform: a multiplex immunoaffinity mass spectrometry platform for protein quantification, and an immunological array platform quantifying glycoprotein isoforms. Submissions provided a mutually beneficial way for members of the proteomics and regulatory communities to identify the analytical issues that the field should address when developing protein-based multiplex clinical assays. PMID- 20007859 TI - Analytical validation of protein-based multiplex assays: a workshop report by the NCI-FDA interagency oncology task force on molecular diagnostics. AB - Clinical proteomics has the potential to enable the early detection of cancer through the development of multiplex assays that can inform clinical decisions. However, there has been some uncertainty among translational researchers and developers as to the specific analytical measurement criteria needed to validate protein-based multiplex assays. To begin to address the causes of this uncertainty, a day-long workshop titled "Interagency Oncology Task Force Molecular Diagnostics Workshop" was held in which members of the proteomics and regulatory communities discussed many of the analytical evaluation issues that the field should address in development of protein-based multiplex assays for clinical use. This meeting report explores the issues raised at the workshop and details the recommendations that came out of the day's discussions, such as a workshop summary discussing the analytical evaluation issues that specific proteomic technologies should address when seeking US Food and Drug Administration approval. PMID- 20007860 TI - Quantification of serum 1-84 parathyroid hormone in patients with hyperparathyroidism by immunocapture in situ digestion liquid chromatography tandem mass spectrometry. AB - BACKGROUND: Immunoassays specific for 1-84 parathyroid hormone (PTH) reportedly reflect the bioactivity of PTH; however, PTH immunoassays can be susceptible to interference by cross-reacting PTH fragments. In addition, these assays currently lack standardization. A methodology using immunocapture purification with liquid chromatography-tandem mass spectrometry (LC-MS/MS) detection, along with a stable isotope-labeled internal standard, may help address these issues. METHODS: We isolated 1-84 PTH from 1 mL serum by immunocapture on a 6.5-mm polystyrene bead. The immobilized PTH was digested in situ and analyzed by LC-MS/MS. For quantification, we used the selected reaction monitoring response from the N terminal tryptic peptide 1-13 PTH ((1)SVSEIQLMHNLGK(13)). RESULTS: The linear range of the assay was 39.1-4560 ng/L, and the limit of detection and limit of quantification were 14.5 ng/L and 39.1 ng/L, respectively. The intraassay CVs ranged from 6% to 11%, and the interassay CVs ranged from 7% to 17%. Interference by PTH fragments 1-44 PTH, 7-84 PTH, 43-68 PTH, 52-84 PTH, 64-84 PTH, and PTH related protein (PTHrP) was 550 men (15 to 49 years) with first ischemic stroke in the Baltimore-Washington area in 2004 to 2007. Frozen plasma samples were assayed for alpha-Gal A activity, and DNA from patients with consistently low plasma alpha-Gal A activities were sequenced. RESULTS: The study sample consisted of 558 men (42% African-American; median age 44 years). Stroke was cryptogenic in 154 men (40% African-American). In 10 patients with low plasma alpha-Gal A activities, DNA sequencing identified alterations in the alpha-Gal A gene in 2 patients. The polymorphism, D313Y, which results in low plasma enzyme activity, but near normal levels of cellular activity was seen in one European-American male. The Fabry disease-causing A143T mutation was seen in an African-American male with cryptogenic stroke (0.18% of all strokes: upper 95% CI=0.53%; 0.65% of cryptogenic strokes: upper 95% CI=1.92%). CONCLUSIONS: In this biracial population, unrecognized Fabry disease is a rare but treatable cause of initial ischemic stroke in young men. PMID- 20007920 TI - Sympathetic control of the cerebral vasculature in humans. AB - BACKGROUND AND PURPOSE: The role of the sympathetic nervous system in cerebral autoregulation remains poorly characterized. We examined cerebral blood flow responses to augmented arterial pressure oscillations with and without sympathetic blockade and compared them with responses in the forearm circulation. METHODS: An oscillatory lower body negative pressure of 40 mm Hg was used at 6 frequencies from 0.03 to 0.08 Hz in 11 healthy subjects with and without alpha adrenergic blockade by phentolamine. RESULTS: Sympathetic blockade resulted in unchanged mean pressure and cerebral flow. The transfer function relationship to arterial pressure at frequencies >0.05 Hz was significantly increased in both the cerebral and brachial circulations, but the coherence of the relation remained weak at the lowest frequencies in the cerebral circulation. CONCLUSIONS: Our data demonstrate a strong, frequency-dependent role for sympathetic regulation of blood flow in both cerebral and brachial circulations. However, marked differences in the response to blockade suggest the control of the cerebral circulation at longer time scales is characterized by important nonlinearities and relies on regulatory mechanisms other than the sympathetic system. PMID- 20007922 TI - Can the genetics of type 1 and type 2 diabetes shed light on the genetics of latent autoimmune diabetes in adults? AB - The pathophysiology of latent autoimmune diabetes in adults (LADA) is considered less understood than its much better characterized counterparts of type 1 and type 2 diabetes (T1D and T2D), where its clinical presentation exhibits some features of each of these two main diseases, earning it a reputation as being "type 1.5 diabetes". The etiology of LADA remains unknown, but a genetic component has been implicated from recent reports of T1D and T2D genes playing a role in its pathogenesis. One way to shed much needed light on the classification of LADA is to determine the discrete genetic factors conferring risk to the pathogenesis of this specific phenotype and to determine to what extent LADA shares genetic similarities with T1D and T2D. For instance, no conclusive support for a role of the T1D-associated INS gene has been reported in T2D; conversely, but similarly, no evidence has been found for the role of the T2D-associated genes IDE/HHEX, SLC30A8, CDKAL1, CDKN2A/B, IGF2BP2, FTO, and TCF7L2 in T1D. However, and somewhat at odds with current thinking, TCF7L2, the most strongly associated gene with T2D to date, is strongly associated with LADA, a disorder considered by the World Health Organization to be a slowly progressing form of T1D. In this review, we address recent advances in the genetics of T1D and T2D and how such discoveries have in turn shed some light on the genetics of LADA as being potentially at the "genetic intersection" of these two major diseases. PMID- 20007921 TI - Cardiotoxicity of anticancer drugs: the need for cardio-oncology and cardio oncological prevention. AB - Due to the aging of the populations of developed countries and a common occurrence of risk factors, it is increasingly probable that a patient may have both cancer and cardiovascular disease. In addition, cytotoxic agents and targeted therapies used to treat cancer, including classic chemotherapeutic agents, monoclonal antibodies that target tyrosine kinase receptors, small molecule tyrosine kinase inhibitors, and even antiangiogenic drugs and chemoprevention agents such as cyclooxygenase-2 inhibitors, all affect the cardiovascular system. One of the reasons is that many agents reach targets in the microenvironment and do not affect only the tumor. Combination therapy often amplifies cardiotoxicity, and radiotherapy can also cause heart problems, particularly when combined with chemotherapy. In the past, cardiotoxic risk was less evident, but it is increasingly an issue, particularly with combination therapy and adjuvant therapy. Today's oncologists must be fully aware of cardiovascular risks to avoid or prevent adverse cardiovascular effects, and cardiologists must now be ready to assist oncologists by performing evaluations relevant to the choice of therapy. There is a need for cooperation between these two areas and for the development of a novel discipline, which could be termed cardio-oncology or onco-cardiology. Here, we summarize the potential cardiovascular toxicities for a range of cancer chemotherapeutic and chemopreventive agents and emphasize the importance of evaluating cardiovascular risk when patients enter into trials and the need to develop guidelines that include collateral effects on the cardiovascular system. We also discuss mechanistic pathways and describe several potential protective agents that could be administered to patients with occult or overt risk for cardiovascular complications. PMID- 20007923 TI - Safety and efficacy of an inhaled epidermal growth factor receptor inhibitor (BIBW 2948 BS) in chronic obstructive pulmonary disease. AB - RATIONALE: Epidermal growth factor receptor (EGFR) activation is implicated in mucin hypersecretion in chronic obstructive pulmonary disease (COPD). OBJECTIVES: To investigate the safety and efficacy of an inhaled EGFR antagonist (BIBW 2948) in COPD. METHODS: Multicenter, double-blind, placebo-controlled trial of 4 weeks of treatment with two doses of BIBW 2948 (15 and 30 mg twice a day) on safety and mucin-related outcomes in 48 patients with COPD. The effect of BIBW 2948 on EGFR activation in airway epithelial cells was assessed using an ex vivo assay. Efficacy measures included the volume of mucin in the airway epithelium (Vs mu,bala) in bronchial biopsies and the expression of mucin genes in bronchial brushings. MEASUREMENTS AND MAIN RESULTS: Inhaled BIBW 2948 induced a dose related inhibition of EGFR internalization (reflecting decreased EGFR activation) in epithelial cells from treated subjects. However, BIBW 2948 was associated with a dose-related increase in adverse events, including reversible liver enzyme elevation (n = 2), and reduction in FEV(1). The changes in mucin stores and mucin gene expression were not significantly different in the pooled BIBW 2948 group versus placebo (volume of mucin per surface area of basal lamina = 0.22 +/- 7.11 vs. 0.47 +/- 8.06 microm(3)/microm(2); P = 0.93). However, in the 30 mg twice a day group, the reduction in epithelial mucin stores was greatest in subjects with the greatest degree of EGFR inhibition (Pearson r = 0.98; 95% confidence interval, 0.71-0.99). CONCLUSIONS: Four-week treatment with BIBW 2948 did not significantly decrease epithelial mucin stores and was poorly tolerated in patients with COPD. Ex vivo analyses suggest that higher doses may be more effective at both EGFR inhibition and decreases in mucin stores but that adverse events should be expected. Clinical trial registered with www.clinicaltrials.gov (NCT00423137). PMID- 20007924 TI - The 15q24/25 susceptibility variant for lung cancer and chronic obstructive pulmonary disease is associated with emphysema. AB - RATIONALE: Genome-wide association studies have identified genetic variants in the nicotinic acetylcholine receptor (nAChR) on chromosome 15q24/25 as a risk for nicotine dependence, lung cancer, and chronic obstructive pulmonary disease (COPD). Assessment of bronchial obstruction by spirometry, typically used for diagnosing COPD, fails, however, to detect emphysema. OBJECTIVES: To determine the association of the 15q24/25 locus with emphysema. METHODS: The rs1051730 variant on 15q24/25 was genotyped in two independent white cohorts of 661 and 456 heavy smokers. Participants underwent pulmonary function tests and computed tomography (CT) of the chest, and took questionnaires assessing smoking behavior and health status. MEASUREMENTS AND MAIN RESULTS: The rs1051730 A-allele correlated with reduced FEV(1) and with increased susceptibility for bronchial obstruction with a pooled odds ratio (OR) of 1.33 (95% confidence interval [CI] = 1.11-1.61; P = 0.0026). In both studies a correlation between the rs1051730 A allele and lung diffusing capacity (Dl(CO)) and diffusing capacity per unit alveolar volume (Kco) was observed. Consistently, the rs1051730 A-allele conferred increased risk for emphysema as assessed by CT (P = 0.0097 and P = 0.019), with a pooled OR of 1.39 (CI = 1.15-1.68; P = 0.00051). Visual emphysema scores and scores based on densities quantified on CT were more pronounced in A allele carriers, indicating that rs1051730 correlates with the severity of emphysema. CONCLUSIONS: The 15q24/25 locus in nAChR is associated with the presence and severity of emphysema. This association was independent of pack years smoking, suggesting that nAChR is causally involved in alveolar destruction as a potentially shared pathogenic mechanism in lung cancer and COPD. PMID- 20007925 TI - Fluorescence in situ hybridization testing algorithm improves lung cancer detection in bronchial brushing specimens. AB - RATIONALE: Bronchoscopically collected cytology specimens are commonly used to obtain a diagnosis of cancer in patients with pulmonary lesions. However, the sensitivity of cytology is suboptimal, especially for peripheral lesions less than 2 cm in diameter. OBJECTIVES: We assessed the performance of a testing algorithm using cytology and fluorescence in situ hybridization (FISH) as part of clinical practice. METHODS: Bronchial brushing specimens (n = 343) were obtained from patients undergoing bronchoscopy for indeterminate pulmonary lesions. Routine cytology was performed and specimens without a positive diagnosis (n = 294) were analyzed by FISH, using residual brushing material. Pathology-confirmed lung cancer or clinical/radiographic evidence of disease was considered diagnostic of malignancy. MEASUREMENTS AND MAIN RESULTS: Routine cytology had a sensitivity and specificity of 41% (23 of 56) and 100% (45 of 45) for central lesions and 20% (26 of 133) and 100% (109 of 109) for peripheral nodules, respectively. FISH detected an additional 32% of lung cancers (18 central and 43 peripheral) not detectable by cytology alone, while producing false positive diagnoses in 22% (10 of 45) and 6% (6 of 109) benign central and peripheral lesions, respectively. In peripheral nodules, FISH detected (relative to routine cytology) an additional 44% (15 of 34) and 28% (25 of 91) of lung cancers less than 2 cm and 2 cm or more in size, respectively. A positive FISH result had a likelihood ratio of 1.45 and 5.87 for central and peripheral lesions and 3.44 and 15.38 for peripheral nodules less than 2 cm and 2 cm or more in size, respectively. CONCLUSIONS: FISH testing significantly increases the detection of lung cancer over routine cytology alone. It is especially useful for peripheral nodules. PMID- 20007926 TI - Increased fraction of exhaled nitric oxide predicts new-onset wheeze in a general population. AB - RATIONALE: Fraction of exhaled nitric oxide (Fe(NO)) is regarded as a marker of airway inflammation. It is unknown whether increased Fe(NO) in respiratorily healthy subjects increases the risk of developing wheeze. OBJECTIVES: To examine if increased levels of Fe(NO) predicts later onset of wheeze. METHODS: We followed up 2,200 men and women from a general population-based study. At baseline, the subjects were investigated with questionnaires, blood samples, pulmonary function tests, and Fe(NO). At follow-up 4 years later, all subjects were mailed a respiratory questionnaire. The association between incident wheeze and baseline levels of Fe(NO) over the 90th percentile were evaluated calculating hazard ratios using Cox regression models adjusted for smoking habits, age, height, sex, and atopy. MEASUREMENTS AND MAIN RESULTS: The follow-up questionnaire was completed by 1,896 subjects (86.2%). All subjects reporting wheeze, asthma, or asthma symptoms at baseline were excluded resulting in a study population of 1,506 subjects. Of these, 49 subjects reported new-onset wheeze. The median concentration of Fe(NO) at baseline was significantly higher among those with new-onset wheeze (18.8 ppb vs. 15.8 ppb, P = 0.03). In a Cox regression model including all subjects, Fe(NO) over the 90th percentile predicted onset of wheeze (hazard ratio 2.7; 95% confidence interval, 1.4-5.4). In stratified models, this was most apparent among never-smokers and in atopic subjects, for whom the odds ratios were higher. CONCLUSIONS: Our results indicate that increased Fe(NO) is associated with an increased risk of developing wheeze. The results also support the hypothesis that increased level of Fe(NO) among subjects without respiratory symptoms is a sign of subclinical airways inflammation. PMID- 20007927 TI - Imatinib treatment for idiopathic pulmonary fibrosis: Randomized placebo controlled trial results. AB - RATIONALE: Idiopathic pulmonary fibrosis (IPF) is a progressive lung disease with no known efficacious therapy. Imatinib is a tyrosine kinase inhibitor with potential efficacy to treat fibrotic lung disease. OBJECTIVES: To investigate the safety and clinical effects of imatinib in patients with IPF. METHODS: We studied 119 patients in an investigator-initiated, multicenter, multinational, double blind clinical trial to receive imatinib or placebo for 96 weeks. MEASUREMENTS AND MAIN RESULTS: Over 96 weeks of follow-up, imatinib did not differ significantly from placebo (log rank P = 0.89) for the primary endpoint defined as time to disease progression (10% decline in percent predicted FVC from baseline) or time to death. There was no effect of imatinib therapy on change in FVC at 48, 72, or 96 weeks (P > or = 0.39 at all time points) or change in diffusing capacity of carbon monoxide at 48, 72, or 96 weeks (P > or = 0.26 at all time points). Change in resting Pa(O(2)) favored imatinib therapy at 48 weeks (P = 0.005) but not at 96 weeks (P = 0.074). During the 96-week trial there were 8 deaths in the imatinib group and 10 deaths in the placebo group (log rank test P = 0.64). Thirty-five (29%) patients discontinued the study without reaching the primary endpoint (imatinib, 32%; placebo, 27%; P = 0.51). Serious adverse events (SAEs) were not more common in the imatinib group (imatinib, 18 SAEs in 17 patients; placebo, 19 SAEs in 18 patients). CONCLUSIONS: In a randomized, placebo controlled trial of patients with mild to moderate IPF followed for 96 weeks, imatinib did not affect survival or lung function. Clinical trial registered with www.clinicaltrials.gov (NCT00131274). PMID- 20007928 TI - Inducible nitric oxide synthase/CD95L-dependent suppression of pulmonary and bone marrow eosinophilia by diethylcarbamazine. AB - RATIONALE: The mechanism of action of diethylcarbamazine (DEC), an antifilarial drug effective against tropical pulmonary eosinophilia, remains controversial. DEC effects on microfilariae depend on inducible NO synthase (iNOS). In eosinophilic pulmonary inflammation, its therapeutic mechanism has not been established. We previously described the rapid up-regulation of bone marrow eosinophilopoiesis in ovalbumin (OVA)-sensitized mice by airway allergen challenge, and further evidenced the down-regulation of eosinophilopoiesis by iNOS- and CD95L-dependent mechanisms. OBJECTIVES: We investigated whether: (1) DEC can prevent the effects of airway challenge of sensitized mice on lungs and bone marrow, and (2) its effectiveness depends on iNOS/CD95L. METHODS: OVA sensitized BALB/c mice were intranasally challenged for 3 consecutive days, with DEC administered over a 12-, 3-, or 2-day period, ending at the day of the last challenge. We evaluated: (1) airway resistance, cytokine (IFN-gamma, IL-4, IL-5, and eotaxin) production, and pulmonary eosinophil accumulation; and (2) bone marrow eosinophil numbers in vivo and eosinophil differentiation ex vivo. MEASUREMENTS AND MAIN RESULTS: DEC effectively prevented the effects of subsequent challenges on: (1) airway resistance, Th1/Th2 cytokine production, and pulmonary eosinophil accumulation; and (2) eosinophilopoiesis in vivo and ex vivo. Recovery from unprotected challenges included full responses to DEC during renewed challenges. DEC directly suppressed IL-5-dependent eosinophilopoiesis in naive bone marrow. DEC was ineffective in CD95L-deficient gld mice and in mice lacking iNOS activity because of gene targeting or pharmacological blockade. CONCLUSIONS: DEC has a strong impact on pulmonary eosinophilic inflammation in allergic mice, as well as on the underlying hemopoietic response, suppressing the eosinophil lineage by an iNOS/CD95L-dependent mechanism. PMID- 20007929 TI - Nontuberculous pulmonary mycobacteriosis in Denmark: incidence and prognostic factors. AB - RATIONALE: Few population-based data are available regarding nontuberculous mycobacteria (NTM) pulmonary disease epidemiology and prognosis. OBJECTIVES: To examine NTM pulmonary colonization incidence, disease incidence, and prognostic factors. METHODS: All adults in Denmark with at least one NTM-positive pulmonary specimen during 1997 to 2008 were identified using national medical databases and were categorized as having possible or definite NTM disease or colonization. MEASUREMENTS AND MAIN RESULTS: We calculated annual age-standardized NTM incidence rates and adjusted hazard ratios (HR) of death associated with patient age, sex, comorbidity, NTM species, and NTM disease status. Of 1,282 adults with 2,666 NTM-positive pulmonary specimens, 335 (26%) had definite NTM disease, 238 (19%) possible disease, and 709 (55%) colonization only. NTM incidence rates decreased until 2002, followed by an increase from 2003 to 2008 (mean annual rate per 100,000 person-years: NTM colonization, 1.36; NTM disease, 1.08). Five-year mortality after definite NTM disease was 40.1%. After controlling for potential confounders, 5-year mortality for definite NTM disease was slightly higher than for NTM colonization (adjusted hazard ratio [HR], 1.15; 95% confidence interval [CI], 0.90-1.51). Mycobacterium xenopi was associated with worse prognosis (adjusted HR, 1.51; 95% CI, 0.99-2.33) than the reference Mycobacterium avium complex. High comorbidity level (HR, 2.97), age greater than or equal to 65 years (HR, 9.17), and male sex (female sex HR, 0.73) were predictors of death. CONCLUSIONS: NTM disease incidence has remained unchanged in Denmark over the past 12 years. Patients with NTM colonization and disease have similarly poor prognosis. Negative prognostic factors include high levels of comorbidity, advanced age, male sex, and M. xenopi. PMID- 20007930 TI - A functional haplotype in the 3'untranslated region of TNFRSF1B is associated with tuberculosis in two African populations. AB - RATIONALE: Susceptibility to tuberculosis is not only determined by Mycobacterium tuberculosis infection, but also by the genetic component of the host. The pleiotropic cytokine tumor necrosis factor-alpha is essential to control tuberculosis infection, and various tumor necrosis factor family members and their respective receptors may contribute to tuberculosis risk. OBJECTIVES: To investigate four functionally relevant polymorphisms in the tumor necrosis factor receptor 2-encoding gene, tumor necrosis factor receptor superfamily member 1B, for association with tuberculosis susceptibility. METHODS: Genotyping of four polymorphisms was performed in independent populations from South Africa (429 cases and 482 control subjects) and Ghana (640 cases and 1,158 control subjects), and the association of the variants with tuberculosis was tested using two case control association studies. MEASUREMENTS AND MAIN RESULTS: Single-point and haplotype analysis in South Africans revealed an association in the 3'untranslated region of the investigated gene. The T allele of rs3397 alone and/or the 3' untranslated region haplotype GTT may confer protection against tuberculosis insofar as both allele and haplotype frequencies were significantly lower in case subjects than in controls. The GTT genotype had previously been shown to increase the decay of tumor necrosis factor receptor 2 messenger ribonucleic acid, and messenger ribonucleic acid destabilization may represent a key molecular mechanism for disease susceptibility. Interestingly, the association signal appeared to be restricted to women. The genetic finding was validated in female participants from Ghana. The combined P value in the haplotype analysis was P = 0.00011. CONCLUSIONS: Our finding emphasizes the importance of tumor necrosis factor/tumor necrosis factor receptor-mediated immune responses in the pathogenesis of tuberculosis. PMID- 20007931 TI - TLR4 is necessary for hyaluronan-mediated airway hyperresponsiveness after ozone inhalation. AB - RATIONALE: Ozone is a common environmental air pollutant that contributes to hospitalizations for respiratory illness. The mechanisms, which regulate ozone induced airway hyperresponsiveness, remain poorly understood. We have previously reported that toll-like receptor 4 (TLR4)-deficient animals are protected against ozone-induced airway hyperresponsiveness (AHR) and that hyaluronan (HA) mediates ozone-induced AHR. However, the relation between TLR4 and hyaluronan in the airway response to ozone remains unexplored. OBJECTIVES: We hypothesized that HA acts as an endogenous TLR4 ligand for the development of AHR after ozone-induced environmental airway injury. METHODS: TLR4-deficient and wild-type C57BL/6 mice were exposed to either inhaled ozone or intratracheal HA and the inflammatory and AHR response was measured. MEASUREMENTS AND MAIN RESULTS: TLR4-deficient mice have similar levels of cellular inflammation, lung injury, and soluble HA levels as those of C57BL/6 mice after inhaled ozone exposure. However, TLR4-deficient mice are partially protected from AHR after ozone exposure as well as after direct intratracheal instillation of endotoxin-free low molecular weight HA. Similar patterns of TLR4-dependent cytokines were observed in the bronchial alveolar lavage fluid after exposure to either ozone or HA. Exposure to ozone increased immunohistological staining of TLR4 on lung macrophages. Furthermore, in vitro HA exposure of bone marrow-derived macrophages induced NF-kappaB and production of a similar pattern of proinflammatory cytokines in a manner dependent on TLR4. CONCLUSIONS: Our observations support the observation that extracellular matrix HA contributes to ozone-induced airways disease. Furthermore, our results support that TLR4 contributes to the biological response to HA by mediating both the production of proinflammatory cytokines and the development of ozone-induced AHR. PMID- 20007932 TI - Long-term effect of continuous positive airway pressure in hypertensive patients with sleep apnea. AB - RATIONALE: Continuous positive airway pressure (CPAP) is the current treatment for patients with symptomatic obstructive sleep apnea (OSA). Its use for all subjects with sleep-disordered breathing, regardless of daytime symptoms, is unclear. OBJECTIVES: This multicenter controlled trial assesses the effects of 1 year of CPAP treatment on blood pressure (BP) in nonsymptomatic, hypertensive patients with OSA. METHODS: We evaluated 359 patients with OSA. Inclusion criteria consisted of an apnea-hypopnea index (AHI) greater than 19 hour(-1), an Epworth Sleepiness Scale score less than 11, and one of the following: under antihypertensive treatment or systolic blood pressure greater than 140 or diastolic blood pressure greater than 90 mm Hg. Patients were randomized to CPAP (n = 178) or to conservative treatment (n = 181). BP was evaluated at baseline and at 3, 6, and 12 months of follow-up. MEASUREMENTS AND MAIN RESULTS: Mean (SD) values were as follows: age, 56 +/- 10 years; body mass index (BMI), 32 +/- 5 kg x m(-2); AHI, 45 +/- 20 hour(-1); and Epworth Sleepiness Scale score, 7 +/- 3. After adjusting for follow-up time, baseline blood pressure values, AHI, time with arterial oxygen saturation less than 90%, and BMI, together with the change in BMI at follow-up, CPAP treatment decreased systolic blood pressure by 1.89 mm Hg (95% confidence interval: -3.90, 0.11 mm Hg; P = 0.0654), and diastolic blood pressure by 2.19 mm Hg (95% confidence interval: -3.46, -0.93 mm Hg; P = 0.0008). The most significant reduction in BP was in patients who used CPAP for more than 5.6 hours per night. CPAP compliance was related to AHI and the decrease in Epworth Sleepiness Scale score. CONCLUSIONS: In nonsleepy hypertensive patients with OSA, CPAP treatment for 1 year is associated with a small decrease in BP. This effect is evident only in patients who use CPAP for more than 5.6 hours per night. Clinical trial registered with www.clinicaltrials.gov (NCT00127348). PMID- 20007933 TI - Palifermin induces alveolar maintenance programs in emphysematous mice. AB - RATIONALE: Emphysema is characterized by destruction of alveoli with ensuing airspace enlargement and loss of alveoli. Induction of alveolar regeneration is still a major challenge in emphysema therapy. OBJECTIVES: To investigate whether therapeutic application of palifermin (DeltaN23-KGF) is able to induce a regenerative response in distal lung parenchyma after induction of pulmonary emphysema. METHODS: Mice were therapeutically treated at three occasions by oropharyngeal aspiration of 10 mg DeltaN23-KGF per kg body weight after induction of emphysema by porcine pancreatic elastase. MEASUREMENTS AND MAIN RESULTS: Airflow limitation associated with emphysema was largely reversed as assessed by noninvasive head-out body plethysmography. Porcine pancreatic elastase-induced airspace enlargement and loss of alveoli were partially reversed as assessed by design-based stereology. DeltaN23-KGF induced proliferation of epithelium, endothelium, and fibroblasts being associated with enhanced differentiation as well as increased expression of vascular endothelial growth factor, vascular endothelial growth factor receptors, transforming growth factor (TGF)-beta1, TGF beta2, (phospho-) Smad2, plasminogen activator inhibitor-1, and elastin as assessed by quantitative reverse transcriptase-polymerase chain reaction, Western blotting, and immunohistochemistry. DeltaN23-KGF induced the expression of TGF beta1 in and release of active TGF-beta1 from primary mouse alveolar epithelial type 2 (AE2) cells, murine AE2-like cells LA-4, and cocultures of LA-4 and murine lung fibroblasts (MLF), but not in MLF cultured alone. Recombinant TGF-beta1 but not DeltaN23-KGF induced elastin gene expression in MLF. Blockade of TGF signaling by neutralizing antibody abolished these effects of DeltaN23-KGF in LA 4/MLF cocultures. CONCLUSIONS: Our data demonstrate that therapeutic application of DeltaN23-KGF has the potential to induce alveolar maintenance programs in emphysematous lungs and suggest that the regenerative effect on interstitial tissue is linked to AE2 cell-derived TGF-beta1. PMID- 20007934 TI - A central role of RLIP76 in regulation of glycemic control. AB - OBJECTIVE: Pathology associated with oxidative stress frequently results in insulin resistance. Glutathione (GSH) and GSH-linked metabolism is a primary defense against oxidative stress. Electrophilic lipid alkenals, such as 4-hydroxy t-2-nonenal (4HNE), generated during oxidative stress are metabolized primarily to glutathione electrophile (GS-E) conjugates. Recent studies show that RLIP76 is the primary GS-E conjugate transporter in cells, and a regulator of oxidative stress response. Because RLIP76(-/-) mice are hypoglycemic, we studied the role of RLIP76 in insulin resistance. RESEARCH DESIGN AND METHODS: Blood glucose, insulin, lipid measurements, and hyperinsulinemic-euglycemic and hyperglycemic clamp experiments were performed in RLIP76(+/+) and RLIP76(-/-) C57B mice, using Institutional Animal Care and Use Committee-approved protocols. Time-resolved three-dimensional confocal fluorescence microscopy was used to study insulin endocytosis. RESULTS: The plasma insulin/glucose ratio was ordered RLIP76(-/-) < RLIP76(+/-) < RLIP76(+/+); administration of purified RLIP76 in proteoliposomes to RLIP76(+/+) animals further increased this ratio. RLIP76 was induced by oxidative or hyperglycemic stress; the concomitant increase in insulin endocytosis was completely abrogated by inhibiting the transport activity of RLIP76. Hydrocortisone could transiently correct hypoglycemia in RLIP76(-/-) animals, despite inhibited activity of key glucocorticoid-regulated hepatic gluconeogenic enzymes, phosphoenolpyruvate carboxykinase, glucose-6-phosphatase, and fructose 1,6-bisphosphatase, in RLIP76(-/-). CONCLUSIONS: The GS-E conjugate transport activity of RLIP76 mediates insulin resistance by enhancing the rate of clathrin-dependent endocytosis of insulin. Because RLIP76 is induced by oxidative stress, it could play a role in insulin resistance seen in pathological conditions characterized by increased oxidative stress. PMID- 20007935 TI - A possible inflammatory role of twist1 in human white adipocytes. AB - OBJECTIVE: Twist1 is a transcription factor that is highly expressed in murine brown and white adipose tissue (WAT) and negatively regulates fatty acid oxidation in mice. The role of twist1 in WAT is not known and was therefore examined. RESEARCH DESIGN AND METHODS: The expression of twist1 was determined by quantitative real-time PCR in different tissues and in different cell types within adipose tissue. The effect of twist1 small interfering RNA on fatty acid oxidation, lipolysis, adipokine secretion, and mRNA expression was determined in human adipocytes. The interaction between twist1 and specific promoters in human adipocytes was investigated by chromatin immunoprecipitation (ChIP) and reporter assays. RESULTS: Twist1 was highly expressed in human WAT compared with a set of other tissues and found predominantly in adipocytes. Twist1 levels increased during in vitro differentiation of human preadipocytes. Gene silencing of twist1 in human white adipocytes had no effect on lipolysis or glucose transport. Unexpectedly, and in contrast with results in mice, twist1 RNA interference reduced fatty acid oxidation. Furthermore, the expression and secretion of the inflammatory factors tumor necrosis factor-alpha, interleukin-6, and monocyte chemoattractant protein-1 were downregulated by twist1 silencing. ChIP and reporter assays confirmed twist1 interaction with the promoters of these genes. CONCLUSIONS: Twist1 may play a role in inflammation of human WAT because it can regulate the expression and secretion of inflammatory adipokines via direct transcriptional effects in white adipocytes. Furthermore, twist1 may, in contrast to findings in mice, be a positive regulator of fatty acid oxidation in human white adipocytes. PMID- 20007936 TI - Insulin gene mutations resulting in early-onset diabetes: marked differences in clinical presentation, metabolic status, and pathogenic effect through endoplasmic reticulum retention. AB - OBJECTIVE: Heterozygous mutations in the human preproinsulin (INS) gene are a cause of nonsyndromic neonatal or early-infancy diabetes. Here, we sought to identify INS mutations associated with maturity-onset diabetes of the young (MODY) or nonautoimmune diabetes in mid-adult life, and to explore the molecular mechanisms involved. RESEARCH DESIGN AND METHODS: The INS gene was sequenced in 16 French probands with unexplained MODY, 95 patients with nonautoimmune early onset diabetes (diagnosed at <35 years) and 292 normoglycemic control subjects of French origin. Three identified insulin mutants were generated by site-directed mutagenesis of cDNA encoding a preproinsulin-green fluorescent protein (GFP) (C peptide) chimera. Intracellular targeting was assessed in clonal beta-cells by immunocytochemistry and proinsulin secretion, by radioimmunoassay. Spliced XBP1 and C/EBP homologous protein were quantitated by real-time PCR. RESULTS: A novel coding mutation, L30M, potentially affecting insulin multimerization, was identified in five diabetic individuals (diabetes onset 17-36 years) in a single family. L30M preproinsulin-GFP fluorescence largely associated with the endoplasmic reticulum (ER) in MIN6 beta-cells, and ER exit was inhibited by approximately 50%. Two additional mutants, R55C (at the B/C junction) and R6H (in the signal peptide), were normally targeted to secretory granules, but nonetheless caused substantial ER stress. CONCLUSIONS: We describe three INS mutations cosegregating with early-onset diabetes whose clinical presentation is compatible with MODY. These led to the production of (pre)proinsulin molecules with markedly different trafficking properties and effects on ER stress, demonstrating a range of molecular defects in the beta-cell. PMID- 20007937 TI - First-trimester follistatin-like-3 levels in pregnancies complicated by subsequent gestational diabetes mellitus. AB - OBJECTIVE To determine whether maternal levels of follistatin-like-3 (FSTL3), an inhibitor of activin and myostatin involved in glucose homeostasis, are altered in the first trimester of pregnancies complicated by subsequent gestational diabetes mellitus (GDM). RESEARCH DESIGN AND METHODS This was a nested case control study of subjects enrolled in a prospective cohort of pregnant women with and without GDM (> or =2 abnormal values on a 100-g glucose tolerance test at approximately 28 weeks of gestation). We measured FSTL3 levels in serum collected during the first trimester of pregnancy. Logistic regression analyses were used to determine the risk of GDM. RESULTS Women who developed GDM (n = 37) had lower first-trimester serum levels of FSTL3 compared with women who did not (n = 127) (median 10,789 [interquartile range 7,013-18,939] vs. 30,670 [18,370-55,484] pg/ml, P < 0.001). When subjects were divided into tertiles based on FSTL3 levels, women with the lowest levels demonstrated a marked increase in risk for developing GDM in univariate (odds ratio 11.2 [95% CI 3.6-35.3]) and multivariate (14.0 [4.1-47.9]) analyses. There was a significant negative correlation between first-trimester FSTL3 levels and approximately 28-week nonfasting glucose levels (r = -0.30, P < 0.001). CONCLUSIONS First-trimester FSTL3 levels are associated with glucose intolerance and GDM later in pregnancy. PMID- 20007938 TI - Association of the TNF-{alpha}-C-857T polymorphism with resistance to the cholesterol-lowering effect of HMG-CoA reductase inhibitors in type 2 diabetic subjects. AB - OBJECTIVE An association of the C-857T polymorphism of the tumor necrosis factor alpha (TNF-alpha) gene promoter region with LDL cholesterol levels has been reported. This study was designed to evaluate the relationship between the TNF alpha-C-857T polymorphism and LDL cholesterol levels according to statin treatment in subjects with type 2 diabetes. RESEARCH DESIGN AND METHODS DNA was obtained from 322 Japanese subjects (160 male and 162 female) with type 2 diabetes, and TNF-alpha-C-857T polymorphisms were determined by direct sequencing. Serum LDL cholesterol was measured by a direct method. RESULTS Although serum LDL cholesterol levels were significantly higher in the T carriers (C/T + T/T) than in the non-T carriers (C/C) (3.14 +/- 0.86 vs. 2.89 +/- 0.75 mmol/l, P < 0.05), there was no difference in LDL cholesterol levels between the non-T carriers and the T carriers in statin-untreated subjects (2.87 +/- 0.73 vs. 2.89 +/- 0.76 mmol/l, NS), whereas in statin-treated subjects, LDL cholesterol levels were significantly higher in the T carriers than in the non-T carriers (3.43 +/- 0.89 vs. 2.90 +/- 0.78 mmol/l, P = 0.0007). There were no differences in HDL cholesterol and triglyceride levels between the non-T carriers and the T carriers in both statin-treated and -untreated subjects. The percent decrease in LDL cholesterol levels after administration of statins was significantly smaller in the T carriers compared with the non-T carriers (27.6 vs. 36.4%, P = 0.031). CONCLUSIONS The mutant allele of the C-857T promoter polymorphism of the TNF alpha gene may predispose to resistance to the LDL cholesterol-lowering effect of statins and could be one of the markers used to predict the efficacy of statins. PMID- 20007939 TI - Dietary patterns and risk for diabetes: the multiethnic cohort. AB - OBJECTIVE The high diabetes incidence among Japanese Americans and Native Hawaiians cannot be explained by BMI. Therefore, we examined the influence of three dietary patterns of "fat and meat," "vegetables," and "fruit and milk" on diabetes risk in the Hawaii component of the Multiethnic Cohort with 29,759 Caucasians, 35,244 Japanese Americans, and 10,509 Native Hawaiians. RESEARCH DESIGN AND METHODS Subjects aged 45-75 years completed a baseline food frequency questionnaire. After 14 years of follow-up, 8,587 subjects with incident diabetes were identified through self-reports or health plan linkages. Risk was assessed using Cox regression stratified by age and adjusted for ethnicity, BMI, physical activity, education, total energy, smoking, alcohol intake, marital status, and hypertension. RESULTS Fat and meat was significantly associated with diabetes risk in men (hazard ratio 1.40 [95% CI 1.23-1.60], P(trend) < 0.0001) and women (1.22 [1.06-1.40], P(trend) = 0.004) when extreme quintiles were compared. Except in Hawaiian women, the magnitude of the risk was similar across ethnic groups although not always significant. After stratification by BMI, fat and meat remained a predictor of disease primarily among overweight men and among overweight Japanese women. Vegetables lowered diabetes risk in men (0.86 [0.77 0.95], P(trend) = 0.004) but not in women, whereas fruit and milk seemed to be more beneficial in women (0.85 [0.76-0.96], P(trend) = 0.005) than in men (0.92 [0.83-1.02], P(trend) = 0.04). CONCLUSIONS Foods high in meat and fat appear to confer a higher diabetes risk in all ethnic groups, whereas the effects of other dietary patterns vary by sex and ethnicity. PMID- 20007940 TI - Development and validation of a questionnaire to assess carbohydrate and insulin dosing knowledge in youth with type 1 diabetes. AB - OBJECTIVE The American Diabetes Association advocates insulin regimens for youth with type 1 diabetes that involve adjusting insulin dose based on carbohydrate intake and blood glucose level. Implementing these regimens requires knowledge about carbohydrate content of foods and subsequent calculations of insulin dose, skills that may be difficult to gauge in practice. Therefore, we sought to develop and validate a questionnaire, the PedCarbQuiz (PCQ), to assess carbohydrate and insulin-dosing knowledge in youth with type 1 diabetes. RESEARCH DESIGN AND METHODS After development by an expert panel, the PCQ was administered to 75 youth with type 1 diabetes or their parents. Reliability was assessed by Cronbach alpha and split-half testing. To assess validity, scores were correlated with A1C, expert assessments, parent educational level, and complexity of insulin regimen. RESULTS PCQ mean score was 87 +/- 9.7% (range 42-98%). Cronbach alpha was 0.88, and correlation of split halves was 0.59 (P < 0.0001). Higher PCQ scores correlated significantly with lower A1C (r = -0.29, P = 0.01) and expert assessments (r = 0.56, P < 0.001). Scores were significantly higher in parents with college degrees than in those without (P = 0.01) and in participants with more complex insulin regimens (P = 0.003). CONCLUSIONS The PCQ is a novel, easily administered instrument to assess knowledge about carbohydrates and insulin dosing calculations. Initial analyses support the reliability and validity of the PCQ. PMID- 20007941 TI - Performance of an A1C and fasting capillary blood glucose test for screening newly diagnosed diabetes and pre-diabetes defined by an oral glucose tolerance test in Qingdao, China. AB - OBJECTIVE The study's goal was to evaluate the performance of A1C and fasting capillary blood glucose (FCG) tests as mass screening tools for diabetes and pre diabetes, as determined by the standard oral glucose tolerance test (OGTT). RESEARCH DESIGN AND METHODS Data from 2,332 individuals aged 35-74 years who participated in a population-based cross-sectional diabetes survey in Qingdao, China, were analyzed. A 2-h 75-g OGTT was used to diagnose diabetes. The performance of A1C and FCG was evaluated against the results of the OGTTs by using receiver operating characteristic curve (ROC) analysis. RESULTS The prevalence of newly diagnosed diabetes and pre-diabetes (impaired fasting glucose and/or impaired glucose tolerance) was 11.9 and 29.5%, respectively. For subjects with newly diagnosed diabetes, the area under the ROC curve was 0.67 for A1C and 0.77 for FCG (P < 0.01) in men and 0.67 and 0.75 (P < 0.01) in women, whereas for pre-diabetes, these values were 0.47 and 0.64 (P < 0.001) in men and 0.51 and 0.65 (P < 0.001) in women. At the optimal A1C cutoff point of > or =5.6% for newly diagnosed diabetes, sensitivities (specificities) were 64.4% (61.6%) for men and 62.3% (63.3%) for women. CONCLUSIONS As a screening tool for newly diagnosed diabetes and pre-diabetes, the FCG measurement performed better than A1C in this general Chinese population. PMID- 20007942 TI - Racial and ethnic differences in an estimated measure of insulin resistance among individuals with type 1 diabetes. AB - OBJECTIVE Insulin resistance is greater in racial/ethnic minorities than in non Hispanic whites (NHWs) for those with and without type 2 diabetes. Because previous research on insulin resistance in type 1 diabetes was limited to NHWs, racial/ethnic variation in an estimated measure of insulin resistance in type 1 diabetes was determined. RESEARCH DESIGN AND METHODS The sample included 79 individuals with type 1 diabetes diagnosed at age <18 years (32.9% NHWs, 46.8% non-Hispanic black [NHB], 7.6% other/mixed, and 12.7% Hispanic) and their families. Estimated glucose disposal rate (eGDR) (milligrams per kilogram per minute; a lower eGDR indicates greater insulin resistance) was calculated using A1C, waist circumference, and hypertension status. RESULTS Mean current age was 13.5 years (range 3.2-32.5) and diabetes duration was 5.7 years (0.1-19.9). eGDR was inversely associated with age. Compared with that in NHWs, age-adjusted eGDR was significantly lower among nonwhites (NHB, other/mixed, and Hispanic: Delta = 1.83, P = 0.0006). Age-adjusted eGDR was negatively associated with body fat, triglycerides, urinary albumin/creatinine, acanthosis nigricans, parental obesity, and parental insulin resistance and positively related to HDL and sex hormone-binding globulin. In multivariable analysis, lower eGDR was significantly associated with older age, nonwhite race/ethnicity, acanthosis, and lower HDL. CONCLUSIONS Minorities with type 1 diabetes are significantly more insulin resistant, as measured by eGDR, than NHWs. Exploring potential mechanisms, including disparities in care and/or physiological variation, may contribute to preventing racial/ethnic differences in insulin resistance-associated outcomes. PMID- 20007943 TI - Is there a link between components of health-related functioning and incident impaired glucose metabolism and type 2 diabetes? The Australian Diabetes Obesity and Lifestyle (AusDiab) study. AB - OBJECTIVE: To determine the longitudinal association of components of health related functioning (HRF) with incident impaired glucose metabolism and type 2 diabetes. RESEARCH DESIGN AND METHODS: The Australian Diabetes Obesity and Lifestyle (AusDiab) study is a national, longitudinal study of adults aged > or =25 years from 42 randomly selected areas of Australia. Diabetes status was defined using the World Health Organization criteria, and HRF was assessed using the SF-36 questionnaire in 1999-2000 and 2004-2005. RESULTS: Incident impaired fasting glucose (IFG), impaired glucose tolerance (IGT), and newly diagnosed type 2 diabetes were associated with increased bodily pain at baseline compared with those with normal glucose tolerance (NGT) (IFG P = 0.005, IGT P < 0.004, and newly diagnosed type 2 diabetes P = 0.005), after adjustment. In addition, those with incident IGT and newly diagnosed type 2 diabetes had significantly reduced physical functioning, general health, mental health, and vitality at baseline compared with those with NGT. After we controlled for factors associated with incident diabetes, those in the lowest quartile of the physical component summary scale at baseline had at least a 50% higher risk of progression to impaired glucose metabolism and diabetes 5 years later. CONCLUSIONS: These findings show that incident IFG, IGT, and newly diagnosed type 2 diabetes are associated with reduced HRF independent of cardiovascular disease and that this is evident before the onset of these conditions. If future health promotion campaigns are to effectively target those at high risk of developing diabetes, an understanding of the process of declining health before onset of the disease is essential. PMID- 20007944 TI - Hypoglycemia in type 1 diabetic pregnancy: role of preconception insulin aspart treatment in a randomized study. AB - OBJECTIVE A recent randomized trial compared prandial insulin aspart (IAsp) with human insulin in type 1 diabetic pregnancy. The aim of this exploratory analysis was to investigate the incidence of severe hypoglycemia during pregnancy and compare women enrolled preconception with women enrolled during early pregnancy. RESEARCH DESIGN AND METHODS IAsp administered immediately before each meal was compared with human insulin administered 30 min before each meal in 99 subjects (44 to IAsp and 55 to human insulin) randomly assigned preconception and in 223 subjects (113 for IAsp and 110 for human insulin) randomly assigned in early pregnancy (<10 weeks). NPH insulin was the basal insulin. Severe hypoglycemia (requiring third-party assistance) was recorded prospectively preconception (where possible), during pregnancy, and postpartum. Relative risk (RR) of severe hypoglycemia was evaluated with a gamma frailty model. RESULTS Of the patients, 23% experienced severe hypoglycemia during pregnancy with the peak incidence in early pregnancy. In the first half of pregnancy, the RR of severe hypoglycemia in women randomly assigned in early pregnancy/preconception was 1.70 (95% CI 0.91 3.18, P = 0.097); the RR in the second half of pregnancy was 1.35 (0.38-4.77, P = 0.640). In women randomly assigned preconception, severe hypoglycemia rates occurring before and during the first and second halves of pregnancy and postpartum for IAsp versus human insulin were 0.9 versus 2.4, 0.9 versus 2.4, 0.3 versus 1.2, and 0.2 versus 2.2 episodes per patient per year, respectively (NS). CONCLUSIONS These data suggest that initiation of insulin analog treatment preconception rather than during early pregnancy may result in a lower risk of severe hypoglycemia in women with type 1 diabetes. PMID- 20007945 TI - Minimal contribution of fasting hyperglycemia to the incidence of type 2 diabetes in subjects with normal 2-h plasma glucose. AB - OBJECTIVE To assess the relative contribution of increased fasting and postload plasma glucose concentrations to the incidence of type 2 diabetes in subjects with a normal 2-h plasma glucose concentration. RESEARCH DESIGN AND METHODS A total of 3,450 subjects with 2-h plasma glucose concentration <140 mg/dl at baseline were followed up in the San Antonio Heart Study (SAHS) and the Botnia Study for 7-8 years. The incidence of type 2 diabetes at follow-up was related to the fasting, 1-h, and 2-h plasma glucose concentrations. RESULTS In subjects with 2-h plasma glucose <140 mg/dl, the incidence of type 2 diabetes increased with increasing fasting plasma glucose (FPG) and 1-h and 2-h plasma glucose concentrations. In a multivariate logistic analysis, after adjustment for all diabetes risk factors, the FPG concentration was a strong predictor of type 2 diabetes in both the SAHS and the Botnia Study (P < 0.0001). However, when the 1 h plasma glucose, but not 2-h plasma glucose, concentration was added to the model, FPG concentration was no longer a significant predictor of type 2 diabetes in both studies (NS). When subjects were matched for the level of 1-h plasma glucose concentration, the incidence of type 2 diabetes markedly increased with the increase in 1-h plasma glucose, but the increase in FPG was not associated with a significant increase in the incidence of type 2 diabetes. CONCLUSIONS An increase in postload glycemia in the normal range is associated with an increase in the incidence of type 2 diabetes. After controlling for 1-h plasma glucose concentration, the increase in FPG concentration is not associated with an increase in the incidence of type 2 diabetes. PMID- 20007946 TI - Relationship styles and mortality in patients with diabetes. AB - OBJECTIVE Prior research has shown that less social support is associated with increased mortality in individuals with chronic illnesses. We set out to determine whether lower propensity to seek support as indicated by relationship style, based on attachment theory, is associated with mortality in patients with diabetes. RESEARCH DESIGN AND METHODS A total of 3,535 nondepressed adult patients with type 1 and type 2 diabetes enrolled in a health maintenance organization in Washington State were surveyed at baseline and followed for 5 years. Relationship style was assessed at baseline. Patients with a greater propensity to seek support were classified as having an interactive relationship style and those less inclined to seek support as having an independent relationship style. We collected Washington State mortality data and used Cox proportional hazards models to estimate relative risk (RR) of death for relationship style groups. RESULTS The rate of death in the independent and interactive relationship style groups was 39 and 29 per 1,000 individuals, respectively. Unadjusted RR of death was 1.33 (95% CI 1.12-1.58), indicating an increased risk of death among individuals with an independent relationship style. After adjustment for demographic and clinical covariates, those with an independent relationship style still had a greater risk of death compared with those with an interactive relationship style (hazard ratio 1.20 [95% CI 1.01 1.43]). CONCLUSIONS In a large sample of adult patients with diabetes, a lower propensity to reach out to others is associated with higher mortality over 5 years. Further research is needed to examine possible mechanisms for this relationship and to develop appropriate interventions. PMID- 20007947 TI - Pediatric endocrinologists' management of children with type 2 diabetes. AB - OBJECTIVE To understand physician behaviors and attitudes in managing children with type 2 diabetes. RESEARCH DESIGN AND METHODS A survey was mailed to a nationwide sample of pediatric endocrinologists (PEs). RESULTS A total of 40% of PEs surveyed responded (211 of 527). Concordance with current monitoring guidelines varied widely, ranging from 36% (foot care) to 93% (blood pressure monitoring). Given clinical vignettes addressing hyperlipidemia, hypertension, and microalbuminuria, only 34% of PEs were fully concordant with current treatment guidelines. Reported barriers included concerns about patient adherence, insufficient scientific evidence about treatment, and lack of familiarity with current recommendations. Providers aged < or =45 years or in clinical practice <10 years reported significantly more aggressive management behaviors and had higher concordance with guidelines. CONCLUSIONS Screening and management of pediatric type 2 diabetes varied widely among PEs, suggesting opportunities for quality improvement. More aggressive management of type 2 diabetes among younger providers may be related to recent training when type 2 diabetes was more common. PMID- 20007948 TI - Real-time continuous glucose monitoring in critically ill patients: a prospective randomized trial. AB - OBJECTIVE To evaluate the impact of real-time continuous glucose monitoring (CGM) on glycemic control and risk of hypoglycemia in critically ill patients. RESEARCH DESIGN AND METHODS A total 124 patients receiving mechanical ventilation were randomly assigned to the real-time CGM group (n = 63; glucose values given every 5 min) or to the control group (n = 61; selective arterial glucose measurements according to an algorithm; simultaneously blinded CGM) for 72 h. Insulin infusion rates were guided according to the same algorithm in both groups. The primary end point was percentage of time at a glucose level <110 mg/dl. Secondary end points were mean glucose levels and rate of severe hypoglycemia (<40 mg/dl). RESULTS Percentage of time at a glucose level <110 mg/dl (59.0 +/- 20 vs. 55.0 +/- 18% in the control group, P = 0.245) and the mean glucose level (106 +/- 18 vs. 111 +/- 10 mg/dl in the control group, P = 0.076) could not be improved using real-time CGM. The rate of severe hypoglycemia was lower in the real-time CGM group (1.6 vs. 11.5% in the control group, P = 0.031). CGM reduced the absolute risk of severe hypoglycemia by 9.9% (95% CI 1.2-18.6) with a number needed to treat of 10.1 (95% CI 5.4-83.3). CONCLUSIONS In critically ill patients, real-time CGM reduces hypoglycemic events but does not improve glycemic control compared with intensive insulin therapy guided by an algorithm. PMID- 20007949 TI - Identification of targeted analyte clusters for studies of schizophrenia. AB - The search for biomarkers to diagnose psychiatric disorders such as schizophrenia has been underway for decades. Many molecular profiling studies in this field have focused on identifying individual marker signals that show significant differences in expression between patients and the normal population. However, signals for multiple analyte combinations that exhibit patterned behaviors have been less exploited. Here, we present a novel approach for identifying biomarkers of schizophrenia using expression of serum analytes from first onset, drug-naive patients and normal controls. The strength of patterned signals was amplified by analyzing data in reproducing kernel spaces. This resulted in the identification of small sets of analytes referred to as targeted clusters that have discriminative power specifically for schizophrenia in both human and rat models. These clusters were associated with specific molecular signaling pathways and less strongly related to other neuropsychiatric disorders such as major depressive disorder and bipolar disorder. These results shed new light concerning how complex neuropsychiatric diseases behave at the pathway level and demonstrate the power of this approach in identification of disease-specific biomarkers and potential novel therapeutic strategies. PMID- 20007950 TI - Molecular targets for diabetes mellitus-associated erectile dysfunction. AB - Protein expression profiles in rat corporal smooth muscle tissue were compared between animal models of streptozotocin-induced diabetes mellitus (STZ-DM) and age-matched controls (AMCs) at 1 week and 2 months after induction of hyperglycemia with STZ treatment. At each time point, protein samples from four STZ-DM and four AMC rat corpora tissues were prepared independently and analyzed together across multiple quantitative two-dimensional gels using a pooled internal standard sample to quantify expression changes with statistical confidence. A total of 170 spots were differential expressed among the four experimental groups. A subsequent mass spectrometry analysis of the 170 spots identified a total of 57 unique proteins. Network analysis of these proteins using MetaCore suggested altered activity of transcriptional factors that are of too low abundance to be detected by the two-dimensional gel method. The proteins that were down-regulated with diabetes include isoforms of collagen that are precursors to fibril-forming collagen type 1; Hsp47, which assists and mediates the proper folding of procollagen; and several proteins whose abundance is controlled by sex hormones (e.g. CRP1 and A2U). On the other hand, proteins seen or predicted to be up-regulated include proteins involved in cell apoptosis (e.g. p53, 14-3-3-gamma, Serpinf1, Cct4, Cct5, and Sepina3n), proteins that neutralize the biological activity of nerve growth factor (e.g. anti-NGF 30), and proteins involved in lipid metabolism (e.g. apoA-I and apoA-IV). Subsequent Western blot validation analysis of p53, 14-3-3-gamma, and Hsp47 confirmed increased p53 and 14-3-3-gamma and decreased Hsp47 levels in separate samples. According to the results from the Western blot analysis, Hsp47 protein showed a approximately 3 fold decrease at 1 week and was virtually undetectable at 2 months in diabetic versus control. Taken together, our results identify novel candidate proteins playing a role in erectile dysfunction in diabetes resulting from STZ treatment. PMID- 20007951 TI - Structural analysis of Rtt106p reveals a DNA binding role required for heterochromatin silencing. AB - Rtt106p is a Saccharomyces cerevisiae histone chaperone with roles in heterochromatin silencing and nucleosome assembly. The molecular mechanism by which Rtt106p engages in chromatin dynamics remains unclear. Here, we report the 2.5 A crystal structure of the core domain of Rtt106p, which adopts an unusual "double pleckstrin homology" domain architecture that represents a novel structural mode for histone chaperones. A histone H3-H4-binding region and a novel double-stranded DNA-binding region have been identified. Mutagenesis studies reveal that the histone and DNA binding activities of Rtt106p are involved in Sir protein-mediated heterochromatin formation. Our results uncover the structural basis of the diverse functions of Rtt106p and provide new insights into its cellular roles. PMID- 20007952 TI - Tailoring the length of antiviral treatment for hepatitis C. PMID- 20007953 TI - Transfusion-refractory anaemia in liver cirrhosis. PMID- 20007954 TI - B cells, IgE and mechanisms of type I hypersensitivity in eosinophilic oesophagitis. PMID- 20007955 TI - Predicting the response to infliximab from trough serum levels. PMID- 20007956 TI - Screening for colorectal cancer: flexible sigmoidoscopy and faecal occult blood immunochemical testing. Which test is superior? PMID- 20007957 TI - Optimum use of diuretics in managing ascites in patients with cirrhosis. PMID- 20007958 TI - A case of idiopathic portal vein thrombosis? PMID- 20007959 TI - Dendritic cells in chronic viral hepatitis B and C: victims or guardian angels? PMID- 20007960 TI - TRP channels: new targets for visceral pain. PMID- 20007961 TI - Comment on: prospective randomised multicentre trial comparing the clinical efficacy, safety and patient acceptability of circular stapled anopexy with closed diathermy haemorrhoidectomy. PMID- 20007962 TI - Should we refer diminutive polyps to post-CTC polypectomy? PMID- 20007963 TI - Comparison of staining methods in collagenous colitis. PMID- 20007965 TI - Quality assurance measures in rectal cancer: caveat utilitor. PMID- 20007966 TI - Arabidopsis auxin response factor6 and 8 regulate jasmonic acid biosynthesis and floral organ development via repression of class 1 KNOX genes. AB - Two mutations in Arabidopsis thaliana, auxin response factor6 (arf6) and arf8, concomitantly delayed the elongation of floral organs and subsequently delayed the opening of flower buds. This phenotype is shared with the jasmonic acid (JA) deficient mutant dad1, and, indeed, the JA level of arf6 arf8 flower buds was decreased. Among JA biosynthetic genes, the expression level of DAD1 (DEFECTIVE IN ANTHER DEHISCENCE1) was markedly decreased in the double mutant, suggesting that ARF6 and ARF8 are required for activation of DAD1 expression. The double mutant arf6 arf8 also showed other developmental defects in flowers, such as aberrant vascular patterning and lack of epidermal cell differentiation in petals. We found that class 1 KNOX genes were expressed ectopically in the developing floral organs of arf6 arf8, and mutations in any of the class 1 KNOX genes (knat2, knat6, bp and hemizygous stm) partially suppressed the defects in the double mutant. Furthermore, ectopic expression of the STM gene caused a phenotype similar to that of arf6 arf8, including the down-regulation of DAD1 expression. These results suggested that most defects in arf6 arf8 are attributable to abnormal expression of class 1 KNOX genes. The expression of AS1 and AS2 was not affected in arf6 arf8 flowers, and as1 and arf6 arf8 additively increased the expression of class 1 KNOX genes. We concluded that ARF6 and ARF8, in parallel with AS1 and AS2, repress the class 1 KNOX genes in developing floral organs to allow progression of the development of these organs. PMID- 20007967 TI - SIZ1 controls cell growth and plant development in Arabidopsis through salicylic acid. AB - The post-translational conjugation of small ubiquitin-related modifiers (SUMOs) to other proteins is involved in regulation of many processes in eukaryotic development; although its role in plant development is beginning to be dissected. Previously, we demonstrated that the siz1 mutant, which is impaired in SUMO E3 ligase, showed a dwarf-like shoot phenotype with accumulation of salicylic acid (SA), and the expression of nahG, a bacterial salicylate hydroxylase that catabolizes SA, in siz1 reduced the SA level and suppressed dwarfism. Herein, we provide evidence that the SIZ1 gene controls cell division and elongation through regulation of the SA level. Mature siz1-2 and siz1-3 plants exhibited a dwarf like shoot phenotype that is attributable to decreased leaf cell volume and number relative to the wild type. Cell division and expansion defects caused by siz1 were also suppressed by the expression of nahG. Expression of XTH8 and XTH31, encoding xyloglucan endotransglycosylase/hydrolase, which are thought to facilitate leaf cell expansion, was down-regulated in siz1 leaves. However, reduced XTH8 and XTH31 expression in siz1 plants was restored in nahG siz1-2 plants. These results indicate that SIZ1 regulates cell growth and plant development with regulation of SA accumulation. Also, XTH8 and XTH31 genes may be responsible for reduced leaf cell expansion. PMID- 20007968 TI - CHM-1, a new vascular targeting agent, induces apoptosis of human umbilical vein endothelial cells via p53-mediated death receptor 5 up-regulation. AB - CHM-1 (2'-fluoro-6,7-methylenedioxy-2-phenyl-4-quinolone) has been identified as a potent antitumor agent in human hepatocellular carcinoma; however, its role in tumor angiogenesis is unclear. This study investigated the effects of CHM-1 and the mechanisms by which it exerts its antiangiogenic and vascular disrupting properties. Using a xenograft model antitumor assay, we found that CHM-1 significantly inhibits tumor growth and microvessel formation. Flow cytometry, immunofluorescence microscopy, and cell death enzyme-linked immunosorbent assay kit revealed that CHM-1 inhibits growth of human umbilical vein endothelial cells (HUVEC) by induction of apoptotic cell death in a concentration-dependent manner. CHM-1 also suppresses HUVEC migration and capillary-like tube formation. We were able to correlate CHM-1-induced apoptosis in HUVEC with the cleavage of procaspase-3, -7, and -8, as well as with the cleavage of poly(ADP-ribose) polymerase by Western blotting assay. Such sensitization was achieved through up regulation of death receptor 5 (DR5) but not DR4 or Fas. CHM-1 was also capable of increasing the expression level of p53, and most importantly, the induction of DR5 by CHM-1 was abolished by p53 small interfering RNA. Taken together, the results of this study indicate that CHM-1 exhibits vascular targeting activity associated with the induction of DR5-mediated endothelial cell apoptosis through p53 up-regulation, which suggests its potential as an antivascular and antitumor therapeutic agent. PMID- 20007969 TI - Protein 4.2 binds to the carboxyl-terminal EF-hands of erythroid alpha-spectrin in a calcium- and calmodulin-dependent manner. AB - Spectrin and protein 4.1 cross-link F-actin protofilaments into a network called the membrane skeleton. Actin and 4.1 bind to one end of beta-spectrin. The adjacent end of alpha-spectrin, called the EF-domain, is calmodulin-like, with calcium-dependent and calcium-independent EF-hands. It has no known function. However, the sph(1J)/sph(1J) mouse has very fragile red cells and lacks the last 13 amino acids in the EF-domain, suggesting the domain is critical for skeletal integrity. Using pulldown binding assays, we find the alpha-spectrin EF-domain either alone or incorporated into a mini-spectrin binds native and recombinant protein 4.2 at a previously identified region of 4.2 (G(3) peptide). Native 4.2 binds with an affinity comparable with other membrane skeletal interactions (K(d) = 0.30 microM). EF-domains bearing the sph(1J) mutation are inactive. Binding of protein 4.2 to band 3 (K(d) = 0.45 microM) does not interfere with the spectrin 4.2 interaction. Spectrin-4.2 binding is amplified by micromolar concentrations of Ca(2+) (but not Mg(2+)) by three to five times. Calmodulin also binds to the EF-domain (K(d) = 17 microM), and Ca(2+)-calmodulin blocks Ca(2+)-dependent binding of protein 4.2 but not Ca(2+)-independent binding. The data suggest that protein 4.2 is located near protein 4.1 at the spectrin-actin junctions. Because proteins 4.1 and 4.2 also bind to band 3, the erythrocyte anion channel, we suggest that one or both of these proteins cause a portion of band 3 to localize near the spectrin-actin junctions and provide another point of attachment between the membrane skeleton and the lipid bilayer. PMID- 20007970 TI - Nuclear factor-kappaB (NF-kappaB) is a novel positive transcriptional regulator of the oncogenic Wip1 phosphatase. AB - The nuclear factor-kappaB (NF-kappaB) family of transcription factors plays a key role in inflammation and augments the initiation, promotion, and progression of cancer. NF-kappaB activation generally leads to transcriptional enhancement of genes important in cell survival and cell growth, which is exploited in cancer cells. In this study, we identify an additional oncogene, PPM1D, which encodes for Wip1, as a transcriptional target of NF-kappaB in breast cancer cells. Inhibition of NF-kappaB or activation of NF-kappaB resulted in decreased or increased Wip1 expression, respectively, at both the mRNA and protein levels. PPM1D promoter activity was positively regulated by NF-kappaB, and this regulation was dependent on the presence of the conserved kappaB site in the PPM1D promoter region. Chromatin immunoprecipitation analysis showed basal binding of the p65 NF-kappaB subunit to the PPM1D promoter region encompassing the kappaB site, which is enhanced after NF-kappaB activation by tumor necrosis factor-alpha. Finally, we show that Wip1 expression is induced in lipopolysaccharide-stimulated mouse splenic B-cells and is required for maximum proliferation. Taken together, these data suggest an additional mechanism by which NF-kappaB may promote tumorigenesis, support the selective use of NF-kappaB inhibitors as chemotherapeutic agents for the treatment of human cancers, and further define a function for Wip1 in inflammation. PMID- 20007971 TI - Glycogen synthase kinase 3beta interaction protein functions as an A-kinase anchoring protein. AB - A-kinase anchoring proteins (AKAPs) include a family of scaffolding proteins that target protein kinase A (PKA) and other signaling proteins to cellular compartments and thereby confine the activities of the associated proteins to distinct regions within cells. AKAPs bind PKA directly. The interaction is mediated by the dimerization and docking domain of regulatory subunits of PKA and the PKA-binding domain of AKAPs. Analysis of the interactions between the dimerization and docking domain and various PKA-binding domains yielded a generalized motif allowing the identification of AKAPs. Our bioinformatics and peptide array screening approaches based on this signature motif identified GSKIP (glycogen synthase kinase 3beta interaction protein) as an AKAP. GSKIP directly interacts with PKA and GSK3beta (glycogen synthase kinase 3beta). It is widely expressed and facilitates phosphorylation and thus inactivation of GSK3beta by PKA. GSKIP contains the evolutionarily conserved domain of unknown function 727. We show here that this domain of GSKIP and its vertebrate orthologues binds both PKA and GSK3beta and thereby provides a mechanism for the integration of PKA and GSK3beta signaling pathways. PMID- 20007972 TI - Spectroscopic and functional characterizations of cyanobacterium Synechocystis PCC 6803 mutants on and near the heme axial ligand of cytochrome b559 in photosystem II. AB - The functional role of cytochrome (cyt) b(559) in photosystem II (PSII) was investigated in H22K alpha and Y18S alpha cyt b(559) mutants of the cyanobacterium Synechocystis sp. PCC6803. H22K alpha and Y18S alpha cyt b(559) mutant carries one amino acid substitution on and near one of heme axial ligands of cyt b(559) in PSII, respectively. Both mutants grew photoautotrophically, assembled stable PSII, and exhibited the normal period-four oscillation in oxygen yield. However, both mutants showed several distinct chlorophyll a fluorescence properties and were more susceptible to photoinhibition than wild type. EPR results indicated the displacement of one of the two axial ligands to the heme of cyt b(559) in H22K alpha mutant reaction centers, at least in isolated reaction centers. The maximum absorption of cyt b(559) in Y18S alpha mutant PSII core complexes was shifted to 561 nm. Y18S alpha and H22K alpha mutant PSII core complexes contained predominately the low potential form of cyt b(559). The findings lend support to the concept that the redox properties of cyt b(559) are strongly influenced by the hydrophobicity and ligation environment of the heme. When the cyt b(559) mutations placed in a D1-D170A genetic background that prevents assembly of the manganese cluster, accumulation of PSII is almost completely abolished. Overall, our data support a functional role of cyt b(559) in protection of PSII under photoinhibition conditions in vivo. PMID- 20007973 TI - Pattern recognition receptors require N-glycosylation to mediate plant immunity. AB - N-Glycans attached to the ectodomains of plasma membrane pattern recognition receptors constitute likely initial contact sites between plant cells and invading pathogens. To assess the role of N-glycans in receptor-mediated immune responses, we investigated the functionality of Arabidopsis receptor kinases EFR and FLS2, sensing bacterial translation elongation factor Tu (elf18) and flagellin (flg22), respectively, in N-glycosylation mutants. As revealed by binding and responses to elf18 or flg22, both receptors tolerated immature N glycans induced by mutations in various Golgi modification steps. EFR was specifically impaired by loss-of-function mutations in STT3A, a subunit of the endoplasmic reticulum resident oligosaccharyltransferase complex. FLS2 tolerated mild underglycosylation occurring in stt3a but was sensitive to severe underglycosylation induced by tunicamycin treatment. EFR accumulation was significantly reduced when synthesized without N-glycans but to lesser extent when underglycosylated in stt3a or mutated in single amino acid positions. Interestingly, EFR(N143Q) lacking a single conserved N-glycosylation site from the EFR ectodomain accumulated to reduced levels and lost the ability to bind its ligand and to mediate elf18-elicited oxidative burst. However, EFR-YFP protein localization and peptide:N-glycosidase F digestion assays support that both EFR produced in stt3a and EFR(N143Q) in wild type cells correctly targeted to the plasma membrane via the Golgi apparatus. These results indicate that a single N glycan plays a critical role for receptor abundance and ligand recognition during plant-pathogen interactions at the cell surface. PMID- 20007974 TI - Extracellular high mobility group box-1 (HMGB1) inhibits enterocyte migration via activation of Toll-like receptor-4 and increased cell-matrix adhesiveness. AB - Toll-like receptor-4 (TLR4) is the receptor for bacterial lipopolysaccharide, yet it may also respond to a variety of endogenous molecules. Necrotizing enterocolitis (NEC) is the leading cause of death from gastrointestinal disease in newborn infants and is characterized by intestinal mucosal destruction and impaired enterocyte migration due to increased TLR4 signaling on enterocytes. The endogenous ligands for TLR4 that lead to impaired enterocyte migration remain unknown. High mobility group box-1 (HMGB1) is a DNA-binding protein that is released from injured cells during inflammation. We thus hypothesize that extracellular HMGB1 inhibits enterocyte migration via activation of TLR4 and sought to define the pathways involved. We now demonstrate that murine and human NEC are associated with increased intestinal HMGB1 expression, that serum HMGB1 is increased in murine NEC, and that HMGB1 inhibits enterocyte migration in vitro and in vivo in a TLR4-dependent manner. This finding was unique to enterocytes as HMGB1 enhanced migration of inflammatory cells in vitro and in vivo. In seeking to understand the mechanisms involved, TLR4-dependent HMGB1 signaling increased RhoA activation in enterocytes, increased phosphorylation of focal adhesion kinase, and increased phosphorylation of cofilin, resulting in increased stress fibers and focal adhesions. Using single cell force traction microscopy, the net effect of HMGB1 signaling was a TLR4-dependent increase in cell force adhesion, accounting for the impaired enterocyte migration. These findings demonstrate a novel pathway by which TLR4 activation by HMGB1 delays mucosal repair and suggest a novel potential therapeutic target in the amelioration of intestinal inflammatory diseases like NEC. PMID- 20007975 TI - Protein kinase C zeta mediates cigarette smoke/aldehyde- and lipopolysaccharide induced lung inflammation and histone modifications. AB - Atypical protein kinase C (PKC) zeta is an important regulator of inflammation through activation of the nuclear factor-kappaB (NF-kappaB) pathway. Chromatin remodeling on pro-inflammatory genes plays a pivotal role in cigarette smoke (CS) and lipopolysaccharide (LPS)-induced abnormal lung inflammation. However, the signaling mechanism whereby chromatin remodeling occurs in CS- and LPS-induced lung inflammation is not known. We hypothesized that PKCzeta is an important regulator of chromatin remodeling, and down-regulation of PKCzeta ameliorates lung inflammation by CS and LPS exposures. We determined the role and molecular mechanism of PKCzeta in abnormal lung inflammatory response to CS and LPS exposures in PKCzeta-deficient (PKCzeta(-/-)) and wild-type mice. Lung inflammatory response was decreased in PKCzeta(-/-) mice compared with WT mice exposed to CS and LPS. Moreover, inhibition of PKCzeta by a specific pharmacological PKCzeta inhibitor attenuated CS extract-, reactive aldehydes (present in CS)-, and LPS-mediated pro-inflammatory mediator release from macrophages. The mechanism underlying these findings is associated with decreased RelA/p65 phosphorylation (Ser(311)) and translocation of the RelA/p65 subunit of NF-kappaB into the nucleus. Furthermore, CS/reactive aldehydes and LPS exposures led to activation and translocation of PKCzeta into the nucleus where it forms a complex with CREB-binding protein (CBP) and acetylated RelA/p65 causing histone phosphorylation and acetylation on promoters of pro-inflammatory genes. Taken together, these data suggest that PKCzeta plays an important role in CS/aldehyde- and LPS-induced lung inflammation through acetylation of RelA/p65 and histone modifications via CBP. These data provide new insights into the molecular mechanisms underlying the pathogenesis of chronic inflammatory lung diseases. PMID- 20007977 TI - Membrane anchor R9AP potentiates GTPase-accelerating protein activity of RGS11 x Gbeta5 complex and accelerates inactivation of the mGluR6-G(o) signaling. AB - The R7 subfamily of RGS proteins critically regulates neuronal G protein signaling pathways that are essential for vision, nociception, motor coordination, and reward processing. A member of the R7 RGS family, RGS11, is a GTPase-accelerating protein specifically expressed in retinal ON-bipolar cells where it forms complexes with the atypical G protein beta subunit, Gbeta(5), and transmembrane protein R9AP. Association with R9AP has been shown to be critical for the proteolytic stability of the complex in the retina. In this study we report that R9AP can in addition stimulate the GTPase-accelerating protein activity of the RGS11 x Gbeta(5) complex at Galpha(o). Single turnover GTPase assays reveal that R9AP co-localizes RGS11 x Gbeta(5) and Galpha(o) on the membrane and allosterically potentiates the GTPase-accelerating function of RGS11 x Gbeta(5). Reconstitution of mGluR6-Galpha(o) signaling in Xenopus oocytes indicates that RGS11 x Gbeta(5)-mediated GTPase acceleration in this system requires co-expression of R9AP. The results provide new insight into the regulation of mGluR6-Galpha(o) signaling by the RGS11 x Gbeta(5) x R9AP complex and establish R9AP as a general GTPase-accelerating protein activity regulator of R7 RGS complexes. PMID- 20007976 TI - Liver X receptor agonists augment human islet function through activation of anaplerotic pathways and glycerolipid/free fatty acid cycling. AB - Recent studies in rodent models suggest that liver X receptors (LXRs) may play an important role in the maintenance of glucose homeostasis and islet function. To date, however, no studies have comprehensively examined the role of LXRs in human islet biology. Human islets were isolated from non-diabetic donors and incubated in the presence or absence of two synthetic LXR agonists, TO-901317 and GW3965, under conditions of low and high glucose. LXR agonist treatment enhanced both basal and stimulated insulin secretion, which corresponded to an increase in the expression of genes involved in anaplerosis and reverse cholesterol transport. Furthermore, enzyme activity of pyruvate carboxylase, a key regulator of pyruvate cycling and anaplerotic flux, was also increased. Whereas LXR agonist treatment up-regulated known downstream targets involved in lipogenesis, we observed no increase in the accumulation of intra-islet triglyceride at the dose of agonist used in our study. Moreover, LXR activation increased expression of the genes encoding hormone-sensitive lipase and adipose triglyceride lipase, two enzymes involved in lipolysis and glycerolipid/free fatty acid cycling. Chronically, insulin gene expression was increased after treatment with TO-901317, and this was accompanied by increased Pdx-1 nuclear protein levels and enhanced Pdx-1 binding to the insulin promoter. In conclusion, our data suggest that LXR agonists have a direct effect on the islet to augment insulin secretion and expression, actions that should be considered either as therapeutic or unintended side effects, as these agents are developed for clinical use. PMID- 20007978 TI - Cytotype diversity and genome size variation in eastern Asian polyploid Cardamine (Brassicaceae) species. AB - BACKGROUND AND AIMS: Intraspecific ploidy-level variation is an important aspect of a species' genetic make-up, which may lend insight into its evolutionary history and future potential. The present study explores this phenomenon in a group of eastern Asian Cardamine species. METHODS: Plant material was sampled from 59 localities in Japan and Korea, which were used in karyological (chromosome counting) and flow cytometric analyses. The absolute nuclear DNA content (in pg) was measured using propidium iodide and the relative nuclear DNA content (in arbitrary units) was measured using 4,6-diamidino-2-phenylindole fluorochrome. KEY RESULTS: Substantial cytotype diversity was found, with strikingly different distribution patterns between the species. Two cytotypes were found in C. torrentis sensu lato (4x and 8x, in C. valida and C. torrentis sensu stricto, respectively), which displays a north-south geographical pattern in Japan. Hypotheses regarding their origin and colonization history in the Japanese archipelago are discussed. In Korean C. amaraeiformis, only tetraploids were found, and these populations may in fact belong to C. valida. C. yezoensis was found to harbour as many as six cytotypes in Japan, ranging from hexa- to dodecaploids. Ploidy levels do not show any obvious geographical pattern; populations with mixed ploidy levels, containing two to four cytotypes, are frequently observed throughout the range. C. schinziana, an endemic of Hokkaido, has hexa- and octoploid populations. Previous chromosome records are also revised, showing that they are largely based on misidentified material or misinterpreted names. CONCLUSIONS: Sampling of multiple populations and utilization of the efficient flow cytometric approach allowed the detection of large-scale variation in ploidy levels and genome size variation attributable to aneuploidy. These data will be essential in further phylogenetic and evolutionary studies. PMID- 20007979 TI - The PLATO trial: do you believe in magic? AB - The PLATO trial revealed a remarkable advantage of ticagrelor over clopidogrel in ACS patients. Unless the regulatory authorities discover serious flaws with the study, which is unlikely, the drug may substantially change the present landscape of oral antiplatelet therapy, especially in high-risk patients. Despite a somewhat unfavourable safety profile, ticagrelor has a lot of room to compensate for these well-defined side effects based on a documented absolute mortality reduction, solid prevention of MI, and convincing pattern of benefit growing over time. PMID- 20007980 TI - Identification and functional characterization of the NanH extracellular sialidase from Corynebacterium diphtheriae. AB - Corynebacterium diphtheriae, a pathogenic Gram-positive bacterium, contains sialic acids on its cell surface, but no genes related to sialic acid decoration or metabolism have been reported in C. diphtheriae. In the present study, we have identified a putative sialidase gene, nanH, from C. diphtheriae KCTC3075 and characterized its product for enzyme activity. Interestingly, the recombinant NanH protein was secreted as a catalytically active sialidase into the periplasmic space in Escherichia coli, while the short region at its C-terminus was truncated by proteolysis. We reconstructed a truncated NanH protein (His(6) NanH(DeltaN)) devoid of its signal sequence as a mature enzyme fused with the 6xHis tag at the N-terminal region. The purified His(6)-NanH(DeltaN) can cleave alpha-2,3- and alpha-2,6-linked sialic acid from sialic acid-containing substrates. In addition, even though the efficiency was low, the recombinant His(6)-NanH(DeltaN) was able to catalyse the transfer of sialic acid using several sialoconjugates as donor, suggesting that the reversible nature of C. diphtheriae NanH can be used for the synthesis of sialyl oligosaccharides via transglycosylation reaction. PMID- 20007981 TI - Single-dose daptomycin pharmacokinetics in chronic haemodialysis patients. AB - BACKGROUND: Daptomycin has concentration-dependent antibacterial activity against Gram-positive bacteria. Its use is increasing in haemodialysis units. The manufacturer recommends a 4-6-mg/kg dose administered every 48 hrs for patients receiving haemodialysis. However, there are no published data about daptomycin pharmacokinetics and clearance during haemodialysis. The recommended dosing regimen would conflict with asymmetric thrice-weekly haemodialysis, which yields two ~44-hr and one ~68-hr interdialytic periods. This is the first study to evaluate daptomycin pharmacokinetics in haemodialysis patients, assess the extent of daptomycin dialytic removal and model serum concentrations at 44 and 68 hrs. METHODS: Six otherwise healthy subjects on chronic haemodialysis (55.3 +/- 16.1 years old, three females, 66.2 +/- 14.2 kg) received a single 6-mg/kg dose of daptomycin post-haemodialysis infused over 30 minutes. Serial blood samples were collected for ~44 hrs (pre-next haemodialysis) and throughout the subsequent haemodialysis session with a high permeability haemodialyser. Individual pharmacokinetic parameters determined by compartmental analysis were used to model trough serum concentrations at 44 and 68 hrs with 6-, 8- and 10-mg/kg post haemodialysis doses. RESULTS: The haemodialysis session in this trial yielded mean urea and daptomycin reduction ratios of 79.6 +/- 5.8% and 57.6 +/- 9.2%, respectively. Daptomycin half-life was 19.4 +/- 6.5 and 3.8 +/- 1.1 hrs 'off' and 'on haemodialysis', respectively, with minimal rebound 1 hr post-haemodialysis. All modelled trough concentrations at 44 and 68 hrs at all doses exceed typical minimum inhibitory concentration (MIC(90)) values for Staphylococcus aureus and Enterococcus faecalis. CONCLUSIONS: Daptomycin serum concentrations declined by ~50% after a 4-hr haemodialysis session with a high permeability haemodialyser. A 6-mg/kg i.v. post-haemodialysis thrice-weekly dose should result in sufficient pre-haemodialysis daptomycin serum concentrations even after a 68-hr interdialytic period. PMID- 20007982 TI - Cytokine patterns and survival in haemodialysis patients. AB - BACKGROUND: Increased pro-inflammatory cytokine levels are associated with decreased survival. We performed factor analyses to determine if pro-inflammatory and anti-inflammatory cytokines in haemodialysis (HD) patients load onto one or two discrete factors and assessed if patients with a specific pattern of high pro inflammatory cytokines have decreased survival compared to patients with a high anti-inflammatory cytokine pattern. METHODS: We evaluated 231 HD patients and analyzed them based on the three most common cytokine distribution patterns seen: a high pro-inflammatory group, a high anti-inflammatory group and all others. Survival and Cox regression analyses were performed. RESULTS: Factor analyses of individual cytokines showed that they loaded onto a single factor. Sixty-five patients had a pro-inflammatory pattern of high IL-1, IL-6 and TNF-alpha levels and low anti-inflammatory parameters, including IL-2, IL-4, IL-5, IL-12, CH50 and T-cell number. The next most frequent cytokine pattern was found in 20 patients with high levels of anti-inflammatory parameters. The patients with high pro inflammatory cytokines had decreased survival compared to patients without a characteristic cytokine pattern. CONCLUSIONS: Further research is needed to better define the underlying causes of increased inflammation among end-stage renal disease patients and to apply anti-inflammatory therapies that may mitigate adverse effects on patient outcomes. PMID- 20007983 TI - Introduction: 2009 BJA/PGA Supplement: a selection of eight reviews. PMID- 20007984 TI - Point-of-care coagulation testing and transfusion algorithms. AB - Patients with cardiovascular disease have an array of haemostasis disorders that predispose to the development of thrombotic and embolic disease states. These patients are often maintained on anti-thrombotic medication to prevent adverse cardiovascular events. Patients undergoing cardiac surgery also have haemostatic disorders that include their intrinsic disease state, adjunctive medication, and the coagulation disturbances induced by cardiopulmonary bypass. The following review introduces the monitors that are available for monitoring perioperative coagulation, with an emphasis on cardiovascular surgery. Heparin monitors, platelet function monitors for use in transfusion algorithms, and monitoring anti platelet drugs will be discussed. PMID- 20007986 TI - Obesity, obstructive sleep apnoea, and diabetes mellitus: anaesthetic implications. AB - Obesity is an epidemic in much of the Western World. The extent of this problem, combined with the increasing preference for ambulatory surgical procedures, has produced a difficult situation for many anaesthesiologists. Even the simplest anaesthetic procedures can become very complicated and potentially difficult in this population. Although there are numerous complications associated with obesity, perhaps obstructive sleep apnoea (OSA) and diabetes mellitus are among the more significant. Patients with OSA are often not ideal candidates for certain day-case procedures, but many outpatient procedures can be performed on patients with OSA as long as attention is paid to anaesthetic technique. Diabetic patients are prone to numerous complications in the perioperative period, including cardiac problems, but with careful management, they are able to undergo day-case surgical procedures safely. PMID- 20007987 TI - Near-infrared spectroscopy as an index of brain and tissue oxygenation. AB - Continuous real-time monitoring of the adequacy of cerebral perfusion can provide important therapeutic information in a variety of clinical settings. The current clinical availability of several non-invasive near-infrared spectroscopy (NIRS) based cerebral oximetry devices represents a potentially important development for the detection of cerebral ischaemia. In addition, a number of preliminary studies have reported on the application of cerebral oximetry sensors to other tissue beds including splanchnic, renal, and spinal cord. This review provides a synopsis of the mode of operation, current limitations and confounders, clinical applications, and potential future uses of such NIRS devices. PMID- 20007988 TI - Perioperative visual loss: what do we know, what can we do? AB - Perioperative visual loss (POVL), a rare, but devastating complication, can follow non-ocular surgery. Highest rates of visual loss are with cardiac and spine surgery. The main causes of visual loss after non-ocular surgery are retinal vascular occlusion and ischaemic optic neuropathy. This review updates readers on the incidence, suspected risk factors, diagnosis, and treatment of POVL due to these conditions. PMID- 20007990 TI - Maternal haemorrhage. AB - Maternal haemorrhage is the leading cause of preventable maternal death worldwide and encompasses antepartum, intrapartum, and postpartum bleeding. This review highlights factors that predispose to severe bleeding, its management, and the most recent treatment and guidelines. Advances in obstetric care have provided physicians with the diagnostic tools to detect, anticipate, and prevent severe life-threatening maternal haemorrhage in most patients who have had prenatal care. In an optimal setting, patients at high risk for haemorrhage are referred to tertiary care centres where multidisciplinary teams are prepared to care for and deal with known potential complications. However, even with the best prenatal care, unexpected haemorrhage occurs. The first step in management is stabilization of haemodynamic status, which involves securing large bore i.v. access, invasive monitoring, and aggressive fluid management and transfusion therapy. Care for the patient with maternal bleeding should follow an algorithm that goes through a rapid and successive sequence of medical and surgical approaches to stem bleeding and decrease morbidity and mortality. With the addition of potent uterotonic agents and the advent of minimally invasive interventional radiological techniques such as angiographic embolization and arterial ligation, definitive yet conservative management is now possible in an attempt to avoid hysterectomy in patients with severe peripartum bleeding. If these interventions are inadequate to control the bleeding, the decision to proceed to hysterectomy must be made expeditiously. Recombinant factor VIIa is a relatively new treatment that could prove useful for severe coagulopathy and intractable bleeding. PMID- 20007989 TI - Postoperative delirium and cognitive dysfunction. AB - Postoperative delirium and cognitive dysfunction (POCD) are topics of special importance in the geriatric surgical population. They are separate entities, whose relationship has yet to be fully elucidated. Although not limited to geriatric patients, the incidence and impact of both are more profound in geriatric patients. Delirium has been shown to be associated with longer and more costly hospital course and higher likelihood of death within 6 months or postoperative institutionalization. POCD has been associated with increased mortality, risk of leaving the labour market prematurely, and dependency on social transfer payments. Here, we review their definitions and aetiology, and discuss treatment and prevention in elderly patients undergoing major non-cardiac surgery. Good basic care demands identification of at-risk patients, awareness of common perioperative aggravating factors, simple prevention interventions, recognition of the disease states, and basic treatments for patients with severe hyperactive manifestations. PMID- 20007991 TI - Perioperative considerations for the patient with asthma and bronchospasm. AB - The incidence of asthma is increasing worldwide, but morbidity and mortality are decreasing because of improvements in medical care. Although the incidence of severe perioperative bronchospasm is relatively low in asthmatics undergoing anaesthesia, when it does occur it may be life-threatening. The keys to an uncomplicated perioperative course are assiduous attention to detail in preoperative assessment, and maintenance of the anti-inflammatory and bronchodilatory regimens through the perioperative period. Potential trigger agents should be identified and avoided. Many routinely used anaesthetic agents have an ameliorative effect on airway constriction. Nonetheless, acute bronchospasm can still occur, especially at induction and emergence, and should be promptly and methodically managed. PMID- 20007992 TI - Lung separation and the difficult airway. AB - Selective collapse of a lung and one-lung ventilation (OLV) is now performed for most thoracic surgical procedures. Modern double-lumen endobronchial tubes and bronchial blockers have made lung separation safe and relatively easy to achieve. However, OLV in the patient with a 'difficult airway' can present a challenge to the anaesthesiologist. This review considers the different techniques used to achieve lung separation and their application to the patient with a difficult airway. PMID- 20007993 TI - C-reactive protein and retinal microvascular caliber in a multiethnic asian population. AB - Retinal microvascular caliber is a risk marker for cardiovascular disease. The authors examined the relation between high-sensitivity C-reactive protein (hsCRP) and retinal microvascular caliber in a multiethnic Asian population (n = 3,583) of Chinese, Malays, and Indians aged 24-95 years residing in Singapore (2003 2007). Retinal arteriolar and venular diameters were measured and summarized as central retinal arteriolar equivalent (CRAE) and central retinal venular equivalent (CRVE), respectively. Persons with higher levels of hsCRP had wider CRVE (P(trend) < 0.001). In subgroup analysis stratified for different cardiovascular disease risk factors, the association between hsCRP and CRVE was seen in persons without diabetes (P(trend) < 0.001) but was absent in persons with diabetes (P(trend) = 0.200; P(interaction) = 0.004). No significant interaction between race/ethnicity and hsCRP in relation to retinal vascular caliber was observed. These data suggest that retinal venular caliber is associated with higher levels of hsCRP in Asians, which is consistent with studies in white Caucasian populations, further supporting the concept that retinal venular caliber may be a marker for low-grade systemic inflammation. PMID- 20007994 TI - The association between obesity and low back pain: a meta-analysis. AB - This meta-analysis assessed the association between overweight/obesity and low back pain. The authors systematically searched the Medline (National Library of Medicine, Bethesda, Maryland) and Embase (Elsevier, Amsterdam, the Netherlands) databases until May 2009. Ninety-five studies were reviewed and 33 included in the meta-analyses. In cross-sectional studies, obesity was associated with increased prevalence of low back pain in the past 12 months (pooled odds ratio (OR) = 1.33, 95% confidence interval (CI): 1.14, 1.54), seeking care for low back pain (OR = 1.56, 95% CI: 1.46, 1.67), and chronic low back pain (OR = 1.43, 95% CI: 1.28, 1.60). Compared with non-overweight people, overweight people had a higher prevalence of low back pain but a lower prevalence of low back pain compared with obese people. In cohort studies, only obesity was associated with increased incidence of low back pain for > or =1 day in the past 12 months (OR = 1.53, 95% CI: 1.22, 1.92). Results remained consistent after adjusting for publication bias and limiting the analyses to studies that controlled for potential confounders. Findings indicate that overweight and obesity increase the risk of low back pain. Overweight and obesity have the strongest association with seeking care for low back pain and chronic low back pain. PMID- 20007995 TI - Manufacturing and placing a bespoke support for the Marfan aortic root: description of the method and technical results and status at one year for the first ten patients. AB - Fatal aortic dissection commonly occurs in Marfan syndrome. Prevention currently relies on elective replacement of the aortic root. We are evaluating the placement of a manufactured, bespoke external support derived from a computer aided design in a prospective study. In the first ten patients, measurements were made consistently of the ascending aorta at the level of closure of the aortic valve cusps from magnetic resonance imaging (MRI) studies taken preoperatively and at fixed intervals thereafter. Before and after images were presented for measurement amongst duplicate images of 37 unoperated Marfan patients to permit assessment of intra-observer measurement reproducibility. All images were presented in random sequence to a radiologist unaware of the research question. The largest difference between the preoperative measurement and that made at least one year after surgery was determined. All patients had surgery as planned without complications and were alive at one year. In eight of the ten patients, the largest observed change was a marked reduction in aortic root diameter. The primary objective of this surgery was achieved in each case, reinforcing the ascending aorta whilst leaving the native aortic valve intact and conserving the blood/endothelium interface. PMID- 20007996 TI - Pegfilgrastim versus filgrastim after high-dose chemotherapy and autologous peripheral blood stem cell support. AB - BACKGROUND: American Society of Clinical Oncology guidelines recommend the use of growth factor after high-dose chemotherapy (HDC) and peripheral blood stem cell (PBSC) support. This randomized trial aims to demonstrate the noninferiority of pegfilgrastim (PEG) compared with filgrastim (FIL) after HDC. PATIENTS AND METHODS: Eighty patients were assigned to FIL at a daily dose of 5 mug/kg or a single fixed dose of PEG (6 mg) 1 day after PBSC. The primary end point was the duration of neutropenia both in terms of absolute neutrophil count (ANC) <0.5 x 10(9)/l and of days to reach an ANC >0.5 x 10(9)/l. RESULTS: The mean duration of neutropenia was 6 and 6.2 days and the mean time to reach an ANC >0.5 x 10(9)/l was 11.5 and 10.8 in the FIL and PEG group, respectively. No differences were observed in the mean time to reach an ANC >1.0 x 10(9)/l (12.2 versus 12.0 days) in the incidence of fever (62% versus 56%) and of documented infections (31% versus 25%). The mean duration of antibiotic therapy was 5.7 and 4.0 days in FIL and PEG group, respectively. CONCLUSION: PEG is not inferior to FIL in hematological reconstitution and represents an effective alternative after HDC and PBSC. PMID- 20007997 TI - Nonpegylated liposomal doxorubicin (MyocetTM) combination (R-COMP) chemotherapy in elderly patients with diffuse large B-cell lymphoma (DLBCL): results from the phase II EUR018 trial. AB - BACKGROUND: To evaluate the activity and safety of nonpegylated liposomal doxorubicin (Myocet) when substituted for doxorubicin in the R-CHOP regimen (R COMP). PATIENTS AND METHODS: Seventy-five elderly patients with diffuse large B cell lymphoma (DLBCL) were studied. Only patients with left ventricular ejection fraction (LVEF) > or =50% were allowed. R-COMP regimen was administered every 3 weeks for three cycles, followed by additional five cycles in case of complete response (CR) or partial response. RESULTS: From November 2002 to April 2005, 75 patients were registered, of which 72 were evaluated. Median age was 72 years (range 61-83); 56% of patients had high or high-intermediate International Prognostic Index score. Median LVEF at baseline was 61%. Thirty-eight patients had history of abnormal cardiovascular conditions. The overall response rate was 71%, with a CR rate of 57%. After a median follow-up of 33 months, the 3-year overall survival, failure-free survival, and progression-free survival rates were 72%, 39%, and 69%, respectively. Neutropenia (54%) was the most frequent grade 3 4 adverse event (AE); 21% of patients experienced cardiac AEs, graded as 3-4 in 4% of the cases. CONCLUSION: R-COMP is an effective regimen for the treatment of DLBCL in elderly patients, with an acceptable tolerability profile. PMID- 20007998 TI - Prdm16 is required for normal palatogenesis in mice. AB - Transcriptional cofactors are essential to the regulation of transforming growth factor beta (TGFbeta) superfamily signaling and play critical and widespread roles during embryonic development, including craniofacial development. We describe the cleft secondary palate 1 (csp1) N-ethyl-N-nitrosourea-induced mouse model of non-syndromic cleft palate (NSCP) that is caused by an intronic Prdm16 splicing mutation. Prdm16 encodes a transcriptional cofactor that regulates TGFbeta signaling, and its expression pattern is consistent with a role in palate and craniofacial development. The cleft palate (CP) appears to be the result of micrognathia and failed palate shelf elevation due to physical obstruction by the tongue, resembling human Pierre Robin sequence (PRS)-like cleft secondary palate. PRDM16 should be considered a candidate for mutation in human clefting disorders, especially NSCP and PRS-like CP. PMID- 20008005 TI - Improving outcomes after surgery. PMID- 20008003 TI - Partial agonist activity of the progesterone receptor antagonist RU486 mediated by an amino-terminal domain coactivator and phosphorylation of serine400. AB - Jun dimerization protein-2 (JDP-2) is a progesterone receptor (PR) coregulatory protein that acts by inducing structure and transcriptional activity in the disordered amino-terminal domain (NTD) of PR. JDP-2 can also potentiate the partial agonist activity of the PR antagonist RU486 by mechanisms that have not been defined. Functional mutagenesis experiments revealed that a subregion of the NTD (amino acids 323-427) was required for the partial agonist activity of RU486 induced by PR interaction with JDP-2. However, this subregion was not required for JDP-2 enhancement of the activity of progestin agonists. Mutation of phosphorylation sites within this region of the NTD showed that phosphorylation of serine 400 was required for the partial agonist activity of RU486 stimulated by JDP-2, but was not required for activity of hormone agonist, either in the presence or absence of JDP-2. Cyclin-dependent kinase 2 (Cdk2)/cyclin A is a novel PR coregulator that binds the NTD and acts by phosphorylating steroid receptor coactivator-1 and modulating steroid receptor coactivator-1 interaction with PR. Cdk2/cyclin A also potentiated the partial agonist activity of RU486; however, phosphorylation of serine 400 was not required, indicating that JDP-2 and Cdk2/cyclin A act by distinct mechanisms. We conclude that PR bound to RU486 and associated with JDP-2 adopts an active conformation in a subregion of the NTD requiring phosphorylation of serine 400 that is distinct from that promoted by progestin agonists. These data underscore the structural flexibility of the NTD of PR, and the ability of steroid ligands together with interacting proteins to affect the conformation and activity of the NTD. PMID- 20008006 TI - World hunger: a reasonable proposal. PMID- 20008008 TI - Record number of people cured of tuberculosis in 2008. PMID- 20008007 TI - The association of early IQ and education with mortality: 65 year longitudinal study in Malmo, Sweden. AB - OBJECTIVES: To establish whether differences in early IQ explain why people with longer education live longer, or whether differences in father's or own educational attainment explain why people with higher early IQ live longer. DESIGN: Population based longitudinal study. Mortality risks were estimated with Cox proportional hazards regressions. SETTING: Malmo, Sweden. PARTICIPANTS: 1530 children who took IQ tests at age 10 and were followed up until age 75. RESULTS: Own educational attainment was negatively associated with all cause mortality in both sexes, even when early IQ and father's education were adjusted for (hazard ratio (HR) for each additional year in school 0.91 (95% CI 0.85 to 0.97) for men and HR 0.88 (95 % CI 0.78 to 0.98) for women). Higher early IQ was linked with a reduced mortality risk in men, even when own educational attainment and father's education were adjusted for (HR for one standard deviation increase in IQ 0.85 (95 % CI 0.75 to 0.96)). In contrast, there was no crude effect of early IQ for women, and women with above average IQ had an increased mortality risk when own educational attainment was adjusted for, but only after the age of 60 (HR 1.60 (95 % CI 1.06 to 2.42)). Adding measures of social career over and above educational attainment to the model (for example, occupational status at age 36 and number of children) only marginally affected the hazard ratio for women with above average IQ (<5%). CONCLUSIONS: Mortality differences by own educational attainment were not explained by early IQ. Childhood IQ was independently linked, albeit differently, to male adult mortality and to female adult mortality even when father's education and own educational attainment was adjusted for, thus social background and own social career seem unlikely to be responsible for mortality differences by childhood IQ. The clear difference in the effect of IQ between men and women suggests that the link between IQ and mortality involves the social and physical environment rather than simply being a marker of a healthy body to begin with. Cognitive skills should, therefore, be addressed in our efforts to create childhood environments that promote health. PMID- 20008012 TI - EU calls for uniform action against smoking in public places across all states. PMID- 20008014 TI - Reducing people's vulnerability is key to combating hunger caused by global warming. PMID- 20008016 TI - GPs make deals with local NHS to vaccinate children against swine flu. PMID- 20008018 TI - Patient and parent motivation for orthodontic treatment--a questionnaire study. AB - SUMMARY: The aims of this investigation were to examine patients' and parents/guardians' motivation for seeking orthodontic treatment and to determine the influence of age and gender in this process. The sample comprised 674 subjects (365 girls and 309 boys) aged 7-18 years and 674 of their parents/guardians, as well as 86 adult patients aged 19-42 years (57 females and 29 males). Similar questionnaires, designed to assess motivation for orthodontic treatment, were completed by all subjects. Statistical analysis was undertaken using Pearson's chi-square test. In the patient groups, a desire to improve aesthetics was the main motivational factor for undergoing treatment. With increasing age, 13 per cent more girls (P = 0.039) were more aware of their malocclusion. The influence of their surroundings on the uptake of treatment decreased with increasing age. Less than 5 per cent of the examined subjects started treatment because other children made fun of them, and only 3 per cent of older patients were motivated by future improvements in health. Between 63 and 67 per cent of parents reported pressurizing their children to seek orthodontic treatment in order to avoid possible future claims of neglect. No statistically significant dependence on gender or age of the children was found. Improvement in dental aesthetics was the principle motivational factor for the children (29-48 per cent), their parents/guardians (54 per cent), and adult patients (55 per cent) seeking orthodontic treatment. PMID- 20008019 TI - Secretion and dipeptidyl peptidase-4-mediated metabolism of incretin hormones after a mixed meal or glucose ingestion in obese compared to lean, nondiabetic men. AB - CONTEXT: Glucose-dependent insulinotropic polypeptide (GIP) and glucagon-like peptide-1 (GLP-1) are cleaved by dipeptidyl peptidase-4 (DPP-4); plasma activity of DPP-4 may be increased in obesity. The impact of this increase on incretin hormone secretion and metabolism is not known. OBJECTIVE: The aim of the study was to assess incretin hormone secretion and degradation in lean and obese nondiabetic subjects. DESIGN, SETTINGS, AND PARTICIPANTS: We studied the ingestion of a mixed meal (560 kcal) or oral glucose (2 g/kg) in healthy lean (n = 12; body mass index, 20-25 kg/m(2)) or obese (n = 13; body mass index, 30-35 kg/m(2)) males at a University Clinical Research Unit. MAIN OUTCOME MEASURES: We measured the area under the curve of plasma intact (i) and total (t) GIP and GLP 1 after meal ingestion and oral glucose. RESULTS: Plasma DPP-4 activity was higher in the obese subjects (38.5 +/- 3.0 vs. 26.7 +/- 1.6 mmol/min . microl; P = 0.002). Although GIP secretion (AUC(tGIP)) was not reduced in obese subjects after meal ingestion or oral glucose, AUC(iGIP) was lower in obese subjects (8.5 +/- 0.6 vs. 12.7 +/- 0.9 nmol/liter x 300 min; P < 0.001) after meal ingestion. GLP-1 secretion (AUC(tGLP-1)) was reduced in obese subjects after both meal ingestion (7.3 +/- 0.9 vs. 10.0 +/- 0.6 nmol/liter x 300 min; P = 0.022) and oral glucose (6.6 +/- 0.8 vs. 9.6 +/- 1.1 nmol/liter x 180 min; P = 0.035). iGLP-1 was reduced in parallel to tGLP-1. CONCLUSIONS: 1) Release and degradation of the two incretin hormones show dissociated changes in obesity: GLP-1 but not GIP secretion is lower after meal ingestion and oral glucose, whereas GIP but not GLP 1 metabolism is increased after meal ingestion. 2) Increased plasma DPP-4 activity in obesity is not associated with a generalized augmented incretin hormone metabolism. PMID- 20008020 TI - Intragenic GNAS deletion involving exon A/B in pseudohypoparathyroidism type 1A resulting in an apparent loss of exon A/B methylation: potential for misdiagnosis of pseudohypoparathyroidism type 1B. AB - CONTEXT: Several endocrine diseases that share resistance to PTH are grouped under the term pseudohypoparathyroidism (PHP). Patients with PHP type Ia show additional hormone resistance, defective erythrocyte G(s)alpha activity, and dysmorphic features termed Albright's hereditary osteodystrophy (AHO). Patients with PHP-Ib show less diverse hormone resistance and normal G(s)alpha activity; AHO features are typically absent in PHP-Ib. Mutations affecting G(s)alpha coding exons of GNAS and epigenetic alterations in the same gene are associated with PHP Ia and -Ib, respectively. The epigenetic GNAS changes in familial PHP-Ib are caused by microdeletions near or within GNAS but without involving G(s)alpha coding exons. OBJECTIVE: We sought to identify the molecular defect in a patient who was diagnosed with PHP-Ia based on clinical presentation (hormone resistance and AHO) but displayed the molecular features typically associated with PHP-Ib (loss of methylation at exon A/B) without previously described genetic mutations. METHODS: Microsatellite typing, comparative genome hybridization, and allelic dosage were performed for proband and her parents. RESULTS: Comparative genome hybridization revealed a deletion of 30,431 bp extending from the intronic region between exons XL and A/B to intron 5. The same mutation was also demonstrated, by PCR, in the patient's mother, but polymorphism and allele dosage analyses indicated that she had this mutation in a mosaic manner. CONCLUSION: We discovered a novel multiexonic GNAS deletion transmitted to our patient from her mother who is mosaic for this mutation. The deletion led to different phenotypic manifestations in the two generation and appeared, in the patient, as loss of GNAS imprinting. PMID- 20008021 TI - Secretory type II phospholipase A2 is produced and secreted by epicardial adipose tissue and overexpressed in patients with coronary artery disease. AB - CONTEXT: Epicardial adipose tissue (EAT) is a visceral adipose tissue in close contact with coronary vessels, the excess of which is associated with coronary artery disease (CAD). OBJECTIVE: Our goal was to identify candidate molecule(s) characterizing EAT that could intervene in the pathogenesis of CAD. DESIGN: An approach combining microarrays and bioinformatic sequence analysis tools for predicting secreted proteins (TargetP) was applied to paired biopsies of sc adipose tissue (SAT) and EAT, obtained from patients with or without CAD (NCAD). RESULTS were validated in three independent groups of subjects by quantitative RT PCR, Western blot, immunohistochemistry, and explant secretion. RESULTS: Secretory type II phospholipase A2 (sPLA2-IIA) ranked as the highest gene coding for potentially secreted proteins with the highest overexpression in EAT in both CAD and NCAD. Quantitative RT-PCR confirmed its increased expression in EAT (P < 0.01) as well as EAT from CAD as compared with NCAD (49.3 +/- 13 vs. 17.4 +/- 9.7 P < 0.01). sPLA2-IIA protein levels were higher in EAT than SAT (P < 0.001). EAT explants also showed significantly higher sPLA2-IIA secretion levels than SAT ones (4.37 +/- 2.7 vs. 0.67 +/- 0.28 ng/ml to 1 per gram tissue per 24 h, P < 0.03). sPLA2-IIA labeling was seen in the stroma vascular fraction between adipocytes and in connective capsules in EAT, with no immunostaining of the adipocytes. SAT was weakly labeled following the same process. CONCLUSION: We have shown for the first time an increased expression of sPLA2-IIA in EAT in patients with CAD. sPLA2-IIA is a phospholipase, which has been shown to be an independent risk factor for CAD. These findings suggest that EAT has a potentially pathophysiological role in CAD. PMID- 20008022 TI - Fat mass exerts a greater effect on cortical bone mass in girls than boys. AB - CONTEXT: It is unclear whether fat mass (FM) and lean mass (LM) differ in the way they influence cortical bone development in boys and girls. OBJECTIVE: The aim of the study was to investigate the contributions of total body FM and LM to parameters related to cortical bone mass and geometry. DESIGN/SETTING: We conducted a longitudinal birth cohort study, the Avon Longitudinal Study of Parents and Children. PARTICIPANTS: A total of 4005 boys and girls (mean age, 15.5 yr) participated in the study. OUTCOME MEASURES: We measured cortical bone mass, cortical bone mineral content (BMC(C)), cortical bone mineral density, periosteal circumference (PC), and endosteal circumference by tibial peripheral quantitative computed tomography. RESULTS: LM had a similar positive association with BMC(C) in boys and girls [regression coefficients with 95% confidence interval (CI); P for gender interactions: boys/girls, 0.952 (0.908, 0.997); P = 0.85]. However, the mechanisms by which LM influenced bone mass differed according to gender because LM was positively associated with PC more strongly in girls [boys, 0.579 (0.522, 0.635); girls, 0.799 (0.722, 0.875); P < 0.0001], but was only associated with cortical bone mineral density in boys [boys, 0.443 (0.382, 0.505); girls, 0.014 (-0.070, 0.097); P < 0.0001]. There was a stronger positive association between FM and BMC(C) in girls [boys, 0.227 (0.185, 0.269); girls, 0.355 (0.319, 0.392); P < 0.0001]. This reflected both a greater positive association of FM with PC in girls [boys, 0.213 (0.174, 0.253); girls, 0.312 (0.278, 0.347); P = 0.0002], and a stronger negative association with endosteal circumference(PC) [boys, -0.059 (-0.096, 0.021); girls, -0.181 (-0.215, -0.146); P < 0.0001]. CONCLUSIONS: Whereas LM stimulates the accrual of cortical bone mass to a similar extent in boys and girls, FM is a stronger stimulus for accrual of cortical bone mass in girls, reflecting a greater tendency in females for FM to stimulate periosteal growth and suppress endosteal expansion. PMID- 20008024 TI - Predicting the response to growth hormone treatment in short children with chronic kidney disease. AB - CONTEXT: Short stature in children with chronic kidney disease (CKD) is due to various underlying congenital or acquired renal disorders resulting in variable impairment of renal function and variable response to GH treatment. OBJECTIVE: It was the aim to develop a mathematical model that allows the prediction of the individual growth response and to identify nonresponders. DESIGN: Data from 208 prepubertal children on conservative or dialysis treatment in a large pharmaco epidemiological survey, the KIGS (Pfizer International Growth Database), were used for the model and data from 67 similar CKD patients registered at the Dutch Growth Research Foundation for validation. RESULTS: Annualized height velocity (centimeters per year) during the first year of GH treatment was best predicted by age at start, weight sd score, underlying renal disorder (hereditary kidney disorder), glomerular filtration rate (at baseline), and GH dosage. Using these parameters, the final model explained 37% of the overall variability of growth response. Standard error of the estimates was 1.6 cm. Age was the most important predictor of growth response (20.3% of variability) followed by weight sd score at start, and 27.2% of the variability of the second-year response could be predicted by the first-year response and glomerular filtration rate. Nonresponders of the validation group could be correctly identified. CONCLUSION: Based on simple clinical variables, a robust prediction model was developed that provides realistic expectations of individual growth response to GH in short children with CKD. The model will help in identifying nonresponders and to tailor treatment strategies. PMID- 20008023 TI - Induction of thyroid gene expression and radioiodine uptake in thyroid cancer cells by targeting major signaling pathways. AB - CONTEXT: Radioiodine ablation is commonly used to treat thyroid cancer, but a major challenge is often the loss of radioiodine avidity of the cancer caused by aberrant silencing of iodide-handling genes. OBJECTIVES: This study was conducted to test the therapeutic potential of targeting the aberrantly activated MAPK and PI3K/Akt/mTOR pathways and histone deacetylase to restore radioiodine avidity in thyroid cancer cells. EXPERIMENTAL DESIGN: We tested the effects of specific inhibitors targeting these pathways/molecules that had established clinical applicability, including the MAPK kinase inhibitor RDEA119, mTOR inhibitor temsirolimus, Akt inhibitor perifosine, and histone deacetylase inhibitor SAHA, individually or in combinations, on the expression of iodide-handling genes and radioiodide uptake in a large panel of thyroid cancer cell lines. RESULTS: The expression of a large number of iodide-handling genes could be restored, particularly the sodium/iodide symporter, TSH receptor, and thyroperoxidase, by treating cells with these inhibitors. The effect was particularly robust and synergistic when combinations of inhibitors containing SAHA were used. Robust expression of sodium/iodide symporter in the cell membrane, which plays the most important role in iodide uptake in thyroid cells, was confirmed by immunofluorescent microscopy. Radioiodide uptake by cells was correspondingly induced under these conditions. Thyroid gene expression and radioiodide uptake could both be further enhanced by TSH. CONCLUSIONS: Targeting major signaling pathways could restore thyroid gene expression and radioiodide uptake in thyroid cancer cells. Further studies are warranted to test this therapeutic potential in restoring radioiodine avidity of thyroid cancer cells for effective ablation treatment. PMID- 20008025 TI - Tobacco-use psychosocial risk profiles of girls and boys in urban India: implications for gender-specific tobacco intervention development. AB - INTRODUCTION: This study examined the distribution of psychosocial risk factors and prevalence of tobacco use among youth in urban India by gender. METHODS: Data were obtained from a cross-sectional baseline survey of a group-randomized tobacco intervention trial involving 6th and 8th graders from 32 schools in Delhi and Chennai (N = 11,642). Mixed-effects regression models were used to examine differences in the prevalence of tobacco use by gender, to determine how the relationship between current tobacco use and related psychosocial risk factors varied by gender, to compare the distribution of risk factors by gender, and to determine if any of these relationships varied by grade level or school type. RESULTS: 14.7% of girls and 21.1% of boys reported ever-use of tobacco. The psychosocial risk profile for tobacco use was remarkably similar for boys and girls, though some differences were apparent. For example, exposure to advertising and beliefs about social effects of use were significant risk factors for girls but not for boys. Across the board, girls showed lower risk for all psychosocial risk factors, except for perceived prevalence of chewing and smoking, for which girls had higher risk compared with boys. DISCUSSION: While the psychosocial risk profile for boys suggests a more vulnerable population for tobacco use, the closing gap in tobacco use between boys and girls indicates a need to examine possible differences in psychosocial risk factors. This study reports that there are subtle, but important, differences in risk factors between genders, having implications for gender-specific intervention development. PMID- 20008026 TI - Awareness of age-related change: examination of a (mostly) unexplored concept. AB - This theoretical article discusses the emerging concept of awareness of age related change (AARC). We propose that a focus on AARC extends the research traditions on subjective age experiences and age identity and that examination of this concept can serve a stimulating role in social gerontology. After defining and contrasting AARC against similar concepts, several reasons for the relevance of this mostly unexplored construct are provided. The sample domains of health and physical functioning, cognitive functioning, and interpersonal relations are used to illustrate the relevance of AARC. Based on this review, we then provide a heuristic framework that describes antecedents, processes, and outcomes related to AARC. Overall, we argue that research on AARC should become an integral part of social gerontological research. PMID- 20008027 TI - The influence of family structure on the contact between older parents and their adult biological children and stepchildren in the Netherlands. AB - OBJECTIVES: This article examines the effect that family structure has on the contact between older adults and their (step)children. A comparison is made among 3 family structures: biological families, complex stepfamilies, and simple stepfamilies. METHODS: The sample consists of respondents aged 55 years or older from the "Living Arrangements and Social Networks of Older Adults in the Netherlands" survey of 1992. The contact between biological relationships and steprelationships is measured by means of 2 items: contact frequency and whether contact is perceived as regular and important. RESULTS: Parents have less contact with their biological children in stepfamilies compared with parents with their children in biological families. The contact with biological children is perceived as more often regular and important in biological families and complex stepfamilies compared with simple stepfamilies. No difference was found in the contact between stepparents and stepchildren in simple and complex stepfamilies. However, the contact with stepchildren is perceived as more often regular and important in simple stepfamilies in comparison to complex stepfamilies. DISCUSSION: It is not so much the difference between biological children and stepchildren that counts when studying the contact between (step)parents and (step)children, as what the structure of the aging (step)family is. PMID- 20008029 TI - A qualitative study of lifestyle counselling in general practice in Ireland. AB - BACKGROUND: There is limited information in Ireland on the attitudes of GPs and practice nurses to lifestyle counselling and the strategies or approaches they use. Furthermore, there is no national framework or resources to support the systematic and uniform provision of lifestyle counselling. OBJECTIVES: To explore the views of Irish primary health care practitioners about behavioural risk factor management in particular to the provision of lifestyle counselling. To identify barriers to behavioural risk factor management and to inform the development of a risk factor management toolkit for general practice. METHODS: The research design is a qualitative study consisting of six focus groups with primary health care practitioners in urban and rural locations in the Republic of Ireland. Two focus groups were conducted with GPs, two with practice nurses, one with a mixed group of GPs and practice nurses and one with a Primary Care Team. In total, 56 participants, aged 30-64 years, attended the focus groups. Descriptive analysis was performed. RESULTS: GPs and practice nurses experienced considerable barriers to lifestyle counselling. These include insufficient time, patient resistance, lack of funding for prevention and lack of training. Participants were aware of the value of patient-centred lifestyle counselling; however, the provision of simple lifestyle information and advice was the predominant strategy used. CONCLUSIONS: GPs and practice nurses regularly conduct lifestyle counselling despite considerable barriers. It is essential that they are supported to carry out lifestyle counselling as part of a systematic 'whole practice approach' to prevention in general practice. PMID- 20008028 TI - Does cognitive training improve internal locus of control among older adults? AB - OBJECTIVES: We evaluated the effect of cognitive training among 1,534 participants in the Advanced Cognitive Training for Independent and Vital Elderly (ACTIVE) randomized controlled trial (RCT) on 5-year improvements in 3 cognitive specific measures of locus of control-internal, chance, and powerful others. METHODS: ACTIVE was a multisite RCT (age > or = 65), with 4 groups (memory, reasoning, speed of processing, and no-contact control). Complete 5-year follow up data were available for 1,534 (55%) of the 2,802 participants. A propensity score model was used to adjust for potential attrition bias. Clinically important improvements (and decrements) in the cognitive-specific locus of control scale scores were defined as greater than or equal to 0.5 SD (medium) and greater than or equal to 1.0 SD (large). Multinomial logistic regression was used to simultaneously contrast those who improved and those who declined with those whose locus of control scale score was unchanged. RESULTS: Statistically significant effects reflecting medium-sized (> or = 0.5 SD) improvements in internal locus of control between baseline and the 5-year follow-up were found for the reasoning and speed of processing intervention groups who were 76% (p < .01) and 68% (p < .05) more likely, respectively, to improve than the no-contact control group. No improvement effects were found on the chance or powerful others locus of control measures or for the memory intervention group. CONCLUSION: Cognitive training that targets reasoning and speed of processing can improve the cognitive-specific sense of personal control over one's life in older adults. PMID- 20008030 TI - Altered modulation of intracortical excitability during movement preparation in Gilles de la Tourette syndrome. AB - Gilles de la Tourette syndrome is a neuropsychiatric disorder in which cortical disinhibition has been proposed as a pathophysiological mechanism involved in the generation of tics. Tics are typically reduced during task performance and concentration. How this task-dependent reduction of motor symptoms is represented in the brain is not yet understood. The aim of the current research was to study motorcortical excitability at rest and during the preparation of a simple motor task. Transcranial magnetic stimulation was used to examine corticospinal excitability, short-interval intracortical inhibition and intracortical facilitation in a group of 11 patients with Gilles de la Tourette syndrome and age-matched healthy controls. Parameters of cortical excitability were evaluated at rest and at six points in time during the preparation of a simple finger movement. Patients with Gilles de la Tourette syndrome displayed significantly reduced short-interval intracortical inhibition at rest, while no differences were apparent for unconditioned motor evoked potential or intracortical facilitation. During the premovement phase, significant differences between groups were seen for single pulse motor evoked potential amplitudes and short interval intracortical inhibition. Short-interval intracortical inhibition was reduced in the early phase of movement preparation (similar to rest) followed by a transition towards more inhibition. Subsequently modulation of short-interval intracortical inhibition was comparable to controls, while corticospinal recruitment was reduced in later phases of movement preparation. The present data support the hypothesis of motorcortical disinhibition in Gilles de la Tourette syndrome at rest. During performance of a motor task, patients start from an abnormally disinhibited level of short-interval intracortical inhibition early during movement preparation with subsequent modulation of inhibitory activity similar to healthy controls. We hypothesize that while at rest, abnormal subcortical inputs from aberrant striato-thalamic afferents target the motor cortex, during motor performance, motor cortical excitability most likely underlies top-down control from higher motor areas and prefrontal cortex, which override these abnormal subcortical inputs to guarantee adequate behavioural performance. PMID- 20008031 TI - Enhanced survival of vascular smooth muscle cells accounts for heightened elastin deposition in arteries of neonatal spontaneously hypertensive rats. AB - Abnormal stiffening and narrowing of arteries are characteristic features of spontaneously hypertensive rats (SHR). In this strain, we have previously demonstrated an increased elastin content and abnormal organization of lamellae in conduit and resistance arteries from neonatal rats that preceded the impending inward remodelling, increased vascular stiffness and development of hypertension. The aim of this study was to assess the mechanism responsible for such excessive and aberrant elastin deposition in SHR vessels during perinatal development. We compared elastin, collagen and fibronectin production (inmunocytochemistry and quantitative assay of metabolically labelled insoluble elastin), DNA content as well as cell proliferation (proliferative cellular nuclear antigen, bromodeoxyuridine incorporation) and death rates (propidium iodide exclusion test, terminal transferase nick and labeling (TUNEL) assay) in cultures of vascular smooth muscle cells (VSMC) derived from neonatal SHR and Wistar-Kyoto (WKY) control rats. Cultures of VSMC derived from neonatal SHR exhibited hypertrophy, produced more elastin, collagen and fibronectin and contained more DNA than equally plated WKY counterparts. Further analysis revealed that the higher net DNA content in SHR-derived cultures was due to increased diploidy, but not to a heightened cell multiplication. The SHR-derived VSMC also exhibited lower rates of cell death and apoptosis, which were associated with increased levels of the anti-apoptotic protein, survivin. We therefore conclude that the peculiar heightened survival of matrix-producing VSMC in neonatal SHR is responsible for accumulation of hard-wearing elastin and other extracellular matrix elements in the growing arteries, thereby contributing to the subsequent development of systemic hypertension. PMID- 20008032 TI - The 'neuroadrenergic hypothesis' in hypertension: current evidence. AB - Data collected in experimental animals and in humans support the hypothesis that sympathetic neural mechanisms are involved in the development and progression of hypertension. Direct approaches to assess human adrenergic cardiovascular drive have shown that sympathetic activation occurs in hypertensive patients, the magnitude of which is proportional to the degree of elevation of the blood pressure. Evidence has also been obtained that sympathetic activation participates in the development of hypertension-related target organ damage, such as left ventricular diastolic dysfunction, left ventricular hypertrophy and arterial remodelling and hypertrophy. Despite the large amount of information collected on the main features of the hypertension-related neurogenic abnormality, the causes of the sympathetic activation remain undefined, although alterations in the reflex modulation of adrenergic drive and/or participation of metabolic factors are likely candidates. This paper will provide background information on the behaviour of the sympathetic nervous system in experimental hypertension, followed by a review of the main features, mechanisms and effects of the sympathetic overdrive in human hypertension. Finally, the new frontiers of research in the area of therapeutic intervention aimed at reducing the adrenergic overdrive will be highlighted. PMID- 20008037 TI - Identification of human UGT2B7 as the major isoform involved in the O glucuronidation of chloramphenicol. AB - Chloramphenicol (CP), a broad spectrum antibiotic, is eliminated in humans by glucuronidation. The primary UGT enzymes responsible for CP O-glucuronidation remain unidentified. We have previously identified the 3-O-CP (major) and 1-O-CP (minor) glucuronides by beta-glucuronidase hydrolysis, liquid chromatography tandem mass spectrometry, and 1D/2D H NMR. Reaction phenotyping for the glucuronidation of CP with 12 expressed human liver UGT isoforms has identified UGT2B7 as having the highest activity for 3-O- and 1-O-CP glucuronidation with minor contributions from UGT1A6 and UGT1A9. The kinetics of CP 3-O glucuronidation by pooled human liver microsomes (HLMs) exhibited biphasic Michaelis-Menten kinetics with the apparent high-affinity K(m1) and low-affinity K(m2) values of 46.0 and 1027 microM, whereas expressed UGT2B7 exhibited Michaelis-Menten kinetics with the apparent K(m) value of 109.1 microM. The formation of 1-O-CP glucuronide by pooled HLM and expressed UGT2B7 exhibited substrate inhibition kinetics with apparent K(m) values of 408.2 and 115.0 microM, respectively. Azidothymidine (AZT) and hyodeoxycholic acid (substrates of UGT2B7) inhibited 3-O- and 1-O-CP glucuronidation in pooled HLMs. In 10 donor HLM preparations, both CP 3-O- and CP 1-O-glucuronidation showed a significant correlation with AZT glucuronidation (UGT2B7) (r(s) = 0.85 and r(s) = 0.83, respectively) at 30 microM CP, whereas no significant correlation was observed between CP 3-O-glucuronidation and serotonin glucuronidation (UGT1A6) or propofol glucuronidation (UGT1A9) at this CP concentration. These results suggest that UGT2B7 is the primary human hepatic UDP-glucuronosyltransferase isoform catalyzing 3-O- and 1-O-CP glucuronidation with minor contributions from UGT1A6 and UGT1A9. PMID- 20008033 TI - Myricetin inhibits UVB-induced angiogenesis by regulating PI-3 kinase in vivo. AB - Myricetin is one of the principal phytochemicals in onions, berries and red wine. Previous studies showed that myricetin exhibits potent anticancer and chemopreventive effects. The present study examined the effect of myricetin on ultraviolet (UV) B-induced angiogenesis in an SKH-1 hairless mouse skin tumorigenesis model. Topical treatment with myricetin inhibited repetitive UVB induced neovascularization in SKH-1 hairless mouse skin. The induction of vascular endothelial growth factor, matrix metalloproteinase (MMP)-9 and MMP-13 expression by chronic UVB irradiation was significantly suppressed by myricetin treatment. Immunohistochemical and western blot analyses revealed that myricetin inhibited UVB-induced hypoxia inducible factor-1alpha expression in mouse skin. Western blot analysis and kinase assay data revealed that myricetin suppressed UVB-induced phosphatidylinositol-3 (PI-3) kinase activity and subsequently attenuated the UVB-induced phosphorylation of Akt/p70(S6K) in mouse skin lysates. A pull-down assay revealed the direct binding of PI-3 kinase and myricetin in mouse skin lysates. Our results indicate that myricetin suppresses UVB-induced angiogenesis by regulating PI-3 kinase activity in vivo in mouse skin. PMID- 20008038 TI - Identification of a novel N-carbamoyl glucuronide: in vitro, in vivo, and mechanistic studies. AB - 1-[4-Aminomethyl-4-(3-chlorophenyl)-cyclohexyl]-tetrahydro-pyrimidin- 2-one, 1, was developed as an inhibitor of dipeptidyl peptidase-4 enzyme. Biotransformation studies with 1 revealed the presence of an N-carbamoyl glucuronide metabolite (M1) in rat bile and urine. N-Carbamoyl glucuronides are rarely observed, and little is understood regarding the mechanism of N-carbamoyl glucuronidation. The objectives of the current investigation were to elucidate the structure of the novel N-carbamoyl glucuronide, to investigate the mechanism of N-carbamoyl glucuronide formation in vitro using stable labeled CO(2), UDP glucuronosyltransferase (UGT) reaction phenotyping, and to assess whether M1 was formed to the same extent in vitro across species-mouse, rat, hamster, dog, monkey, and human. Structure elucidation was performed on a mass spectrometer with accurate mass measurement and MS(n) capabilities. (13)C-labeled carbon dioxide was used for identification of the mechanism of N-carbamoyl glucuronidation. Mechanistic studies with (13)C-labeled CO(2) in rat liver microsomes revealed that CO(2) from the bicarbonate buffer (in equilibrium with exogenous CO(2)) may be responsible for the formation of M1. M1 was formed in vitro in liver microsomes from multiple species, mainly rat and hamster, followed by similar formation in dog, monkey, mouse, and human. M1 could be detected in UGT1A1, UGT1A3, and UGT2B7 Supersomes in a CO(2)-rich environment. In conclusion, our study demonstrates that formation of M1 was observed in microsomal incubations across various species and strongly suggests incorporation of CO(2) from the bicarbonate buffer, in equilibrium with exogenous CO(2), into the carbamoyl moiety of the formed N-carbamoyl glucuronide. PMID- 20008039 TI - Glucuronide production by whole-cell biotransformation using genetically engineered fission yeast Schizosaccharomyces pombe. AB - Drug metabolites generated by UDP glycosyltransferases (UGTs) are needed for drug development and toxicity studies, especially in the context of safety testing of metabolites during drug development. Because chemical metabolite synthesis can be arduous, various biological approaches have been developed; however, no whole cell biotransformation with recombinant microbes that express human UGTs was yet achieved. In this study we expressed human UDP glucose-6-dehydrogenase together with several human or rat UGT isoforms in the fission yeast Schizosaccharomyces pombe and generated strains that catalyze the whole-cell glucuronidation of standard substrates. Moreover, we established two methods to obtain stable isotope-labeled glucuronide metabolites: the first uses a labeled aglycon, whereas the second uses (13)C(6)-glucose as a metabolic precursor of isotope labeled UDP-glucuronic acid and yields a 6-fold labeled glucuronide. The system described here should lead to a significant facilitation in the production of both labeled and unlabeled drug glucuronides for industry and academia. PMID- 20008040 TI - How do people make continence care happen? An analysis of organizational culture in two nursing homes. AB - PURPOSE: Although nursing homes (NHs) are criticized for offering poor quality continence care, little is known about the organizational processes that underlie this care. This study investigated the influence of organizational culture on continence care practices in two NHs. DESIGN AND METHODS: This ethnographic study explored continence care from the perspectives of NH stakeholders, including residents and interdisciplinary team members. Data were collected through participant observation, interviews, and archival records. RESULTS: Human relations dimensions of organizational culture influenced continence care by affecting institutional missions, admissions and hiring practices, employee tenure, treatment strategies, interdisciplinary teamwork, and group decision making. Closed system approaches, parochial identity, and an employee focus stabilized staff turnover, fostered evidence-based practice, and supported hierarchical toileting programs in one facility. Within a more dynamic environment, open system approaches, professional identity, and job focus allowed flexible care practices during periods of staff turnover. Neither organizational culture fully supported interdisciplinary team efforts to maximize the bladder and bowel health of residents. IMPLICATIONS: Organizational culture varies in NHs, shaping the continence care practices of interdisciplinary teams and leading to the selective use of treatments across facilities. Human relations dimensions of organizational culture, including open or closed systems, professional or parochial identity, and employee or job focus are critical to the success of quality improvement initiatives. Evidence-based interventions should be tailored to organizational culture to promote adoption and sustainability of resident care programs. PMID- 20008041 TI - Use of mutant mouse lines to investigate origin of gonadotropin-releasing hormone 1 neurons: lineage independent of the adenohypophysis. AB - Mutant mouse lines have been used to study the development of specific neuronal populations and brain structures as well as behaviors. In this report, single- and double-mutant mice were used to examine the lineage of GnRH-1 cells. GnRH is essential for vertebrate reproduction, with either GnRH-1 or GnRH-3 controlling release of gonadotropins from the anterior pituitary, depending on the species. It is clear that the neuroendocrine GnRH cells migrate from extracentral nervous system locations into the forebrain. However, the embryonic origin of GnRH-1 and GnRH-3 cells is controversial and has been suggested to be nasal placode, adenohypophyseal (anterior pituitary) placode, or neural crest, again dependent on the species examined. We found that mutant mice with either missing or disrupted anterior pituitaries (Gli2(-/-), Gli1(-/-)Gli2(-/-), and Lhx3(-/-)) exhibit a normal GnRH-1 neuronal population and that these cells are still found associated with the developing vomeronasal organ. These results indicate that in mice, GnRH-1 cells develop independent of the adenohypophyseal placode and are associated early with the formation of the nasal placode. PMID- 20008042 TI - Oxytocin increases glucose uptake in neonatal rat cardiomyocytes. AB - We have recently shown that an entire oxytocin (OT) system, a peptide and its cognate receptors, is synthesized in the heart. In fetal and newborn hearts, OT exists in its extended three-amino acid form, OT-Gly-Lys-Arg (OT-GKR). OT translocates glucose transporter type 4 to the plasma membrane in human endothelial cells. Therefore, we hypothesized that the cardiac OT/OT-GKR system may be involved in the regulation of myocardial glucose uptake in physiological conditions and during metabolic stress such as hypoxia. Primary cultures of neonatal rat cardiomyocytes (CM) and cardiac progenitor cells expressing ATP binding cassette efflux transporter G2 transporter (stem cell marker) were studied. OT (10 nm) increased basal glucose uptake in CM to 4.0 +/- 0.2 fmol/mg protein, with OT-GKR (10 nm) elevating it to 5.3 +/- 0.4 fmol/mg protein (P < 0.001) in comparison with 2.2 fmol/mg in control cells. OT had a moderate synergistic effect with 0.1 mm 2,4-dinitrophenol, augmenting basal glucose uptake to 5.5 +/- 0.5 fmol/mg. OT-GKR (10 nm) was even more potent in combination with 2,4-dinitrophenol, increasing glucose uptake to 9.0 +/- 1.0 fmol/mg. Wortmannin (0.1 microm), an inhibitor of phosphatidylinositol-3-kinase, significantly suppressed the effect of OT and insulin (10 nm) (P < 0.001), indicating common pathways. Our data suggest that OT and OT-GKR influence glucose uptake in neonatal rat CM and may thus play a role in the maintenance of cardiac function and cell survival during metabolic stress. PMID- 20008043 TI - Risk factors for multidrug-resistant bacteria in patients with post-operative peritonitis requiring intensive care. AB - OBJECTIVES: This prospective non-interventional study investigated the risk factors for multidrug-resistant bacteria (MDRB) in patients with post-operative peritonitis (POP), to provide guidance for empirical antimicrobial therapy. METHODS: All consecutive patients, >15 years old, admitted to a surgical intensive care unit (ICU) between September 2006 and January 2009 for a first episode of POP were included. Antibiotic susceptibilities of microorganisms recovered from blood cultures and peritoneal fluid were determined by disc diffusion. Amoxicillin/clavulanic acid, ticarcillin/clavulanic acid, piperacillin/tazobactam, cefotaxime, ceftazidime, cefepime, imipenem, gentamicin, amikacin and ciprofloxacin were tested against Gram-negative bacteria, and oxacillin, amoxicillin, vancomycin, gentamicin and erythromycin were tested against aerobic Gram-positive bacteria. Results were reported as susceptible or resistant. RESULTS: MDRB were isolated from 20/115 (17%) patients. In univariate analysis, use of antimicrobial therapy during the 3 months prior to hospitalization and a long duration between hospital admission or first operation and relaparotomy were significantly associated with MDRB recovery. In multivariate analysis, only antimicrobial treatment in the 3 months preceding hospitalization and duration between first operation and relaparotomy were independent risk factors for MDRB [odds ratio (OR) = 5.80, 95% confidence interval (95% CI) = 1.99-16.91 and OR = 1.10, 95% CI = 1.02-1.19, respectively]. No MDRB were found when the delay between the first operation and relaparotomy was <5 days. POP severity, non-surgical and surgical complications, hospital and ICU length of stay, and mortality were similar in patients with and without MDRB. CONCLUSIONS: Our results suggest that broad-spectrum antibiotics should be used in ICU patients with POP who have received antimicrobial therapy in the 3 months prior to hospitalization, or with >5 days between the first operation and relaparotomy. PMID- 20008044 TI - Phenotypic and genotypic Helicobacter pylori clarithromycin resistance and therapeutic outcome: benefits and limits. AB - INTRODUCTION: Primary clarithromycin resistance is increasing worldwide, and it has been regarded as the main factor reducing the efficacy of Helicobacter pylori therapy. However, the clinical consequence of either phenotypic or genotypic resistance still remains unclear. This study aimed to evaluate: (i) the concordance between phenotypic (culture) and genotypic (real-time PCR) tests in assessing primary clarithromycin resistance; and (ii) the role of both in therapeutic outcome. METHODS: A post hoc subgroup study was selected from a double-blind, placebo-controlled trial, enrolling 146 patients with dyspepsia or peptic ulcers never previously treated. Real-time PCR and Etest on bacterial culture for assessing clarithromycin resistance were performed. [(13)C]urea breath test (UBT), histology and rapid urease tests at entry and UBT after 4-8 weeks were used to assess infection and eradication. All patients received a 10 day therapy. RESULTS: Prevalence of clarithromycin phenotypic resistance was significantly lower as compared with genotypic resistance (18.4% versus 37.6%, P < 0.001). A concordance between the two methods was present in 71.2% of cases. A significant difference in the eradication rate was seen between clarithromycin susceptible and -resistant strains, when assessed with either Etest (92.4% versus 55.5%, P < 0.001) or a PCR-based method (94.5% versus 70.9%; P < 0.001). Of note, the eradication rate showed the lowest value (30.7%) when phenotypic bacterial resistance was genetically linked to the A2143G point mutation. CONCLUSIONS: This study showed that: (i) there is a relevant discordance between the two methods; and (ii) phenotypic clarithromycin resistance markedly reduces H. pylori eradication when it is linked to a specific point mutation. PMID- 20008045 TI - Detection of a new extended-spectrum oxacillinase in Pseudomonas aeruginosa. PMID- 20008046 TI - Novel role of Acinetobacter baumannii RND efflux transporters in mediating decreased susceptibility to biocides. AB - OBJECTIVES: Biocides and dyes are commonly employed in hospital and laboratory settings. We investigated the biocide susceptibilities of a rapidly emerging pathogen, Acinetobacter baumannii, and the underlying molecular mechanisms, with a primary focus on resistance-nodulation-cell division (RND) efflux systems. METHODS: Biocide susceptibilities, efflux and in vitro inactivation profiles were monitored in the presence/absence of efflux pump inhibitors. The RND transporters encoded by adeB and adeJ were detected by PCR; null mutants were constructed in the native host. Expression of adeB and adeJ in clinical isolates was assayed by semi-quantitative RT-PCR. RESULTS: Susceptibility testing and phenotypic assays demonstrated the role of active efflux in mediating decreased susceptibility to biocides. Inactivation of either the adeB or adeJ transporter gene led to increased susceptibility to biocides. RT-PCR analysis exhibited increased adeB and adeJ expression in clinical isolates. CONCLUSIONS: This is the first study demonstrating the role of efflux pumps in mediating decreased susceptibility to disinfectants and other chemical substrates in A. baumannii. PMID- 20008047 TI - Forty-one recent cases of invasive zygomycosis from a global clinical registry. AB - BACKGROUND: Invasive zygomycosis accounts for a significant proportion of all invasive fungal diseases (IFD), but clinical data on the clinical course and treatment response are limited. PATIENTS AND METHODS: Fungiscope-A Global Rare Fungal Infection Registry is an international university-based case registry that collects data of patients with rare IFD, using a web-based electronic case form at www.fungiscope.net. RESULTS: Forty-one patients with invasive zygomycosis from central Europe and Asia were registered. The most common underlying conditions were malignancies (n = 26; 63.4%), diabetes mellitus (n = 7; 17.1%) and solid organ transplantation (n = 4; 9.8%). Diagnosis was made by culture in 28 patients (68.3%) and by histology in 26 patients (63.4%). The main sites of infection were the lungs (n = 24; 58.5%), soft tissues (n = 8; 19.5%), rhino-sinu-orbital region (n = 8; 19.5%) and brain (n = 6; 14.6%). Disseminated infection of more than one non-contiguous site was seen in six patients (14.6%). Mycocladus corymbifer was the most frequently identified species (n = 10, 24.4%). A favourable response was observed in 23 patients (56.1%). Overall survival was 51.2% (n = 21). At diagnosis, four patients (9.8%) were on continuous antifungal prophylaxis with itraconazole (n = 1; 2.4%) or posaconazole (n = 3; 7.3%). Initial targeted treatment with activity against zygomycetes was administered to 34 patients (82.9%). Liposomal amphotericin B was associated with improved response (P = 0.012) and survival rates (P = 0.004). CONCLUSIONS: Pathogen distribution and, consequently, drug susceptibility seem to vary across different geographic regions. Furthermore, protection from invasive zygomycosis for patients on posaconazole prophylaxis is not absolute. Our findings indicate that the use of liposomal amphotericin B as first-line treatment for patients diagnosed with zygomycoses merits further investigation, preferably in the form of a clinical trial. PMID- 20008048 TI - Resistance development profiling of piperacillin in combination with the novel {beta}-lactamase inhibitor BLI-489. AB - OBJECTIVES: To evaluate development of resistance to the piperacillin/BLI-489 combination. METHODS: BLI-489 was used at a constant concentration of 4 mg/L. Spontaneous mutation frequency was measured on piperacillin/BLI-489-containing agar plates. Five beta-lactamase-producing strains were exposed to a serial dilution of piperacillin/BLI-489, and the highest concentration allowing growth was used to inoculate subsequent serial passage for 10 days. Mutation stability was monitored in drug-free medium for 10 days. RESULTS: Escherichia coli (OXA-3, OXA-7, ACT-1, SHV-1 or none), Salmonella enterica serovar Typhimurium (CTX-M-5), Klebsiella pneumoniae (SHV-1 and SHV-5) and Enterobacter cloacae (AmpC) had a spontaneous mutation frequency of < or =1.0 x 10(-9). Two AmpC-producing Pseudomonas aeruginosa strains had a mutation frequency of 6.52 x 10(-6) and 1.0 x 10(-7); a beta-lactamase-negative P. aeruginosa strain had a mutation frequency of 2.68 x 10(-8). The mutant prevention concentration (MPC) was < or =32 mg/L. During serial passages, the MIC increased 64- and 128-fold for S. enterica serovar Typhimurium (CTX-M-5) and E. cloacae (AmpC), respectively, to > or =512 mg/L. The MIC reverted to < or =64 mg/L after serial passages in drug-free medium. The MICs increased only 4-fold for K. pneumoniae (SHV-1 and SHV-5), E. coli (OXA-3) and E. coli (SHV-1). CONCLUSIONS: Piperacillin/BLI-489 demonstrated a low probability of spontaneous resistance development in vitro for all of the strains tested with the exception of P. aeruginosa. The MPC value for all strains was < or =32 mg/L. Resistance developed during serial passage for two of the five strains tested; however, this resistance phenotype was unstable as MIC values reverted to < or =64 mg/L after propagation in drug-free medium. PMID- 20008049 TI - Variations in colistin susceptibility among different species of the genus Acinetobacter. PMID- 20008050 TI - Climate change and spring-fruiting fungi. AB - Most macrofungi produce ephemeral fruit bodies during autumn but some have adapted to spring fruiting. In this study, temporal changes in the time of spring fruiting in Norway and the UK during 1960-2007 have been investigated by statistical analyses of about 6000 herbarium and field records, covering 34 species. Nearly 30 per cent of the temporal variation in fruiting could be ascribed to spatial and species-specific effects. Correcting for these effects, linear trends towards progressively earlier fruiting were detected during the entire period in both Norway and the UK, with a change in average fruiting day of 18 days over the study period. Early fruiting was correlated with high winter temperatures in both countries, indicating that the observed phenological changes are likely due to earlier onset of spring. There were also significant correlations between climatic conditions in one year and timing of fruiting the following year, indicating that below-ground mycelia are influenced by climatic conditions over a longer time period before fruiting. Fruiting dates were, however, not strictly related to changes in vernal accumulated thermal time. Our results indicate that global warming has lead to progressively earlier fruiting of spring fungi in northwest Europe during the last half century. PMID- 20008051 TI - Home-based speech treatment for Parkinson's disease delivered remotely: a case report. AB - We investigated the validity and feasibility of online delivery of the Lee Silverman Voice Treatment (LSVT) for the treatment of the speech disorder of a patient with idiopathic Parkinson's disease. The treatment was delivered in 16 sessions to the participant's home, 90 km from the speech language pathologist. A PC-based videoconferencing system was used, operating at 128 kbit/s over the public telecommunications network. The patient achieved substantial improvements in vocal sound pressure levels during sustained vowel phonation (6.13 dB), reading (12.28 dB) and conversational monologue (11.32 dB). There were improvements in the duration of sustained vowel phonation (4 s). Improvements were also perceived in the degree of breathiness and roughness in the voice, and in overall speech intelligibility in conversation. The patient was very satisfied with the audio and video quality of the conferencing, and with the online treatment overall. He reported a preference for online sessions for the future management of his condition, rather than face-to-face treatment. Remote LSVT delivery was found to be feasible and effective. PMID- 20008052 TI - Hearing health-care delivery in sub-Saharan Africa--a role for tele-audiology. AB - Hearing loss is the most prevalent chronic disability and a major contributor to the global burden of disease. Its effects are far-reaching and can lead to severely restricted developmental outcomes for children and limited vocational prospects for adults. The benefits of intervention are dramatic and can significantly improve developmental outcomes, especially in infants identified early. Hearing health-care services in developing regions such as sub-Saharan Africa are however severely limited, leaving affected individuals without access to secondary and tertiary intervention. Tele-audiology has potential for improved access to specialist hearing health services including identification, diagnosis and intervention. There has been limited experience so far with tele-audiology in industrialized countries. In the sub-Saharan region, the continued growth in Internet connectivity and general technological advances make tele-audiology an important approach to consider in the management of hearing loss. Pilot studies are warranted to investigate the potential obstacles to the widespread implementation of telehealth in the delivery of hearing health care in poorly resourced communities, in line with World Health Organization initiatives. PMID- 20008053 TI - Cost-effectiveness of a community-based obesity control programme. AB - We evaluated two 12-week long community-based obesity control programmes in Korea. One was a visiting-type programme (V-type) (n = 515) administered by a public health centre and the other was a remote-type programme (R-type) (n = 410) utilizing an Internet website and mobile phones with a short message service. The total cost for the intention-to-treat subjects was US$116,993 in the V-type programme and $24,555 in the R-type programme. In the per-protocol subjects, 66% of V-type participants (n = 117) achieved the target bodyweight reduction (5%) and 13% of R-type participants (n = 15). In the per-protocol subjects, the cost per person was $227 (V-type) and $60 (R-type). The cost per person achieving the target weight reduction was $975 (V-type) and $1637 (R-type). The average amount that participants were willing to pay was $71 (V-type) and $21 (R-type). The cost effectiveness of the visiting-type community-based, short-duration obesity control programme was higher than the remote-type programme. PMID- 20008054 TI - Home telemonitoring for congestive heart failure: a systematic review and meta analysis. AB - We conducted a systematic review of the literature about home telemonitoring compared with usual care. An electronic literature search was conducted to identify studies of home telemonitoring use in congestive heart failure (CHF) patients. Twenty-one original studies on home telemonitoring for patients with CHF were included (3082 patients). A random effects model was used to compute treatment efficacy to measure the average effect of the intervention across all studies where the quantitative pooling of results was appropriate. Home telemonitoring reduced mortality (risk ratio = 0.64; 95% CI: 0.48-0.85) compared with usual care. Several studies suggested that home telemonitoring also helped to lower the number of hospitalizations and the use of other health services. Patient quality of life and satisfaction with home telemonitoring were similar or better than with usual care. More studies of higher methodological quality are required to give more precise information about the potential clinical effectiveness of home telehealth interventions. PMID- 20008055 TI - Telephone-supported care coordination in an Australian veterans population: a randomized controlled trial. AB - An evaluation was undertaken on the effectiveness and efficiency of care coordination in delivering health services to Australian veterans with chronic or complex medical conditions requiring multidisciplinary care and who had moderate to high care needs. The veterans participated in a randomized controlled trial (RCT) supported by the Department of Veterans' Affairs. For evaluation of the RCT, information on cost of care and quality of life (QOL) was collected before the commencement of coordinated care and at follow-up after 12 months. Of 525 veterans who were recruited, 481 were surveyed at baseline (243 in the intervention group and 238 controls). At follow-up, 421 were surveyed (213 intervention and 208 controls). There were no significant differences between the coordinated care and control groups of veterans in costs of care or in QOL measurements using the SF-12 Health Survey and the EuroQol Group EQ-5D. These findings are consistent with those reported in earlier studies which suggest that benefits from care coordination programmes may take some time to emerge. PMID- 20008056 TI - Labour induction with 25 micrograms versus 50 micrograms intravaginal misoprostol in full term pregnancies. AB - We undertook this study in order to compare the efficacy of 25 microg versus 50 microg of intravaginal misoprostol for cervical ripening and labour induction at term. The study population consisted of 120 women with term singleton pregnancies in vertex presentation booked for caesarean section. They had a Bishop scoring of <6 and a reactive fetal heart rate tracing. They were randomized into two groups, A and B, to receive 25 microg and 50 microg of vaginal misoprostol, respectively, 4 hourly with a maximum of five doses until the patient had three contractions in 10 minutes. There was no significant difference in the induction delivery interval between the two groups (12.52+/- 7.05 h in the 25 microg group versus 11.72+/- 6.74 h in the 50 microg group; P = 0.58). Of the women in the 25 microg group, 83.3% delivered vaginally as did 71.67% of those in the 50 microg group, but the difference was not statistically significant (P = 0.128). There were significantly more women requiring oxytocin augmentation in the 25 microg group than in the 50 microg group (P = 0.03). However, there were no significant differences in the rates of caesarean and operative vaginal delivery, meconium stained liquor, fetal distress or in the incidences of hyperstimulation between the two groups. Neonatal outcomes were similar. The intravaginal administrations of 25 microg, as well as 50 microg of misprostol, are equally efficacious in inducing labour. The 25 microg group more often required oxytocin as an adjunct. PMID- 20008057 TI - Evaluation of visual inspection with acetic acid and Lugol's iodine as cervical cancer screening tools in a low-resource setting. AB - In view of the failure of cytology screening programmes for cervical cancer in developing countries, the World Health Organization suggested unaided visual inspection of the cervix after an application of acetic acid (VIA) and Lugol's iodine (VILI) as alternative screening methods. Our study evaluates the effectiveness of VIA and VILI compared to Pap smear as screening methods for carcinoma of the cervix in a low-resource setting. Three hundred and twenty-eight women were subjected to a Pap smear test, VIA, VILI and colposcopy. The results were as follows: Pap smear test (20.83%, specificity 98.38%), VIA (55.5%, 71.39%) and VILI (86.84%, 48.93%). Although VIA and VILI are less specific in comparison to the Pap smear test, they are more sensitive in detecting pre-invasive lesions. Hence VIA and VILI can be used as cervical cancer screening tools in low-resource settings. PMID- 20008058 TI - The role of unorthodox and traditional birth care in maternal mortality. AB - Many pregnant women see unorthodox medical providers in labour before presentation to the modern medical facilities after obstetric complications have arisen. This study evaluates the contribution of unorthodox medical facilities to the delays subsisting maternal mortality in a rural, poor and illiterate community. Data was collected prospectively on all referrals from outside the St. Vincent's hospital, over a three-year period. Seven hundred and fifty women were referred to the hospital and there were a total of thirty maternal deaths out of the 1268 live births, giving a maternal mortality ratio of 2366/100,000. Most of the referrals were patient-driven and verbal and came from traditional birth attendants (TBAs). The majority of the patients (86.7%) came in poor clinical conditions and some were moribund. The TBAs contributed most to maternal deaths. Prolongation of labour for more than 24 hours correlated positively with maternal mortality. Ruptured uterus complicating obstructed labour (34.8%) and haemorrhage (30.4%) were the leading causes of death in this series. The mortal delay suffered by pregnant women in accessing unorthodox medical attention deserves further attention in issues of maternal mortality in the underserved rural communities of Nigeria. PMID- 20008059 TI - Peripartum hysterectomy: a ten-year experience at a tertiary care hospital in a developing country. AB - Acute bleeding after delivery can be a life-threatening complication. Emergency hysterectomy is usually undertaken as a last resort. This study was conducted in order to estimate the incidence, indications, risk factors and complications associated with peripartum hysterectomy performed at a tertiary care hospital. We retrospectively analysed 39 of 45 cases of emergency peripartum hysterectomy performed at the Aga Khan University Hospital from 1997-2006. Peripartum hysterectomy was defined as one performed for a haemorrhage after delivery which is unresponsive to other treatments. The most frequent indications for peripartum hysterectomy were morbidly adherent placenta (46%) and uterine atony (23%). The duration of surgery was shorter (P = 0.045) but the complications were higher (P = 0.029) in total compared with subtotal hysterectomies. Our results suggest that caesarean deliveries are associated with an increased risk for peripartum hysterectomy, which is of concern given the increasing rate of caesarean deliveries. Subtotal hysterectomy is a reasonable alternative in emergency obstetric hysterectomy. PMID- 20008060 TI - Barriers to follow-up of abnormal Papanicolaou smears among women in Leon, Nicaragua. AB - Cervical cancer screening programmes only succeed when the coverage, screening, treatment and follow-up of any abnormal results are of high quality. This study investigates the screening coverage and the quality of the follow-up of cervical cancer screening of women in Leon, Nicaragua, as well as the factors affecting coverage and barriers to follow-up. Two hundred and thirty-four sexually active women attending a public health centre (54), a public health post (63), or the gynaecology department of the public HEODRA hospital (117), were invited to answer an orally administered questionnaire. The screening coverage of cervical cancer was calculated and the follow-up after finding an abnormal screening result was assessed. The coverage was 60.6% (95% confidence interval 54.4-66.8%). One hundred and sixty-eight women (87%) were informed about the results. Eighteen (10, 7%) of the women who were informed about the results, were advised to have follow-up and 12 (66, 7%) of them did. Barriers to follow-up were financial constraints, pregnancy, postponement and absence of instruments/electricity. This study shows that the screening coverage and cytology services are adequate, but that there is insufficient follow-up and/or treatment after an abnormal screening result. In order to achieve a lower incidence and mortality of cervical cancer, follow-up and treatment after an abnormal screening result need to be improved. A visual inspection with acetic acid may be an attractive alternative. PMID- 20008061 TI - Malarial retinopathy in northern Nigerian children. AB - This is a prospective study involving two centres. Children younger than 16 years old who had severe malaria underwent funduscopy. In one centre, direct ophthalmoscopy was performed by both an ophthalmologist and a non-ophthalmologist physician. At the other, two ophthalmologists performed a funduscopy on each patient using different methods (one using direct and the other indirect ophthalmoscopy). The objective was to determine the frequency of retinopathy and evaluate the effectiveness of direct ophthalmoscopy in detecting retinopathy in constantly dilated eyes. Of 62 children seen at both centres, 37% were found to have malarial retinopathy. Macula whitening was the most common finding. There was substantial agreement between the ophthalmologists using either the direct or indirect ophthalmoscopy techniques (Kappa value 0.604). However, at the centre using only direct ophthalmoscopy, there was poor agreement between the ophthalmologist and the non-ophthalmologist (kappa value -0.244). Direct ophthalmoscopy seemed to be effective in detecting malarial retinopathy, but only when performed by experienced personnel. PMID- 20008062 TI - A silybin-phospholipid complex prevents mitochondrial dysfunction in a rodent model of nonalcoholic steatohepatitis. AB - Mitochondrial dysfunction and oxidative stress are determinant events in the pathogenesis of nonalcoholic steatohepatitis. Silybin has shown antioxidant, anti inflammatory, and antifibrotic effects in chronic liver disease. We aimed to study the effect of the silybin-phospholipid complex (SILIPHOS) on liver redox balance and mitochondrial function in a dietary model of nonalcoholic steatohepatitis. To accomplish this, glutathione oxidation, mitochondrial oxygen uptake, proton leak, ATP homeostasis, and H(2)O(2) production rate were evaluated in isolated liver mitochondria from rats fed a methionine- and choline-deficient (MCD) diet and the MCD diet plus SILIPHOS for 7 and 14 weeks. Oxidative proteins, hydroxynonenal (HNE)- and malondialdehyde (MDA)-protein adducts, and mitochondrial membrane lipid composition were also measured. Treatment with SILIPHOS limited glutathione depletion and mitochondrial H(2)O(2) production. Moreover, SILIPHOS preserved mitochondrial bioenergetics and prevented mitochondrial proton leak and ATP reduction. Finally, SILIPHOS limited the formation of HNE- and MDA-protein adducts. In conclusion, SILIPHOS is effective in preventing severe oxidative stress and preserving hepatic mitochondrial bioenergetics in nonalcoholic steatohepatitis induced by the MCD diet. The modifications of mitochondrial membrane fatty acid composition induced by the MCD diet are partially prevented by SILIPHOS, conferring anti-inflammatory and antifibrotic effects. The increased vulnerability of lipid membranes to oxidative damage is limited by SILIPHOS through preserved mitochondrial function. PMID- 20008063 TI - Pulmonary toxicity and metabolic activation of dauricine in CD-1 mice. AB - Dauricine is the major bioactive component isolated from the roots of Menispermum dauricum D.C. and has shown promising pharmacological activities with a great potential for clinic use. However, the adverse effects and toxicity of the alkaloid are unfortunately ignored. The objective of the current study was to evaluate the toxicity of dauricine in vitro and in vivo. Mice (CD-1) were treated intraperitoneally with dauricine at various doses, and sera and lung lavage fluids were collected after 24 h of treatment. No changes in serum aspartate aminotransferase, alanine aminotransferase, and blood urea nitrogen were noticed, whereas a dose-dependent increase in lactate dehydrogenase activity was observed in lung lavage fluids. Ethidium-based staining studies showed that remarkable cells lost membrane integrity in the lungs of the animals treated with dauricine at 150 mg/kg. Histopathological evaluation of lungs of mice showed that dauricine at the same dose caused significant alveolar edema and hemorrhage. Exposure to dauricine at 40 muM for 24 h resulted in up to 60% cell death in human lung cell lines BEAS-2B, WI-38, and A549. Ketoconazole showed protective effect on the pulmonary injury in mice given dauricine. A quinone methide metabolite of dauricine was identified in mouse lung microsomal incubations, and the presence of ketoconazole in the microsomal incubations suppressed the formation of the quinone methide metabolite. In conclusion, dauricine produced pulmonary injury in CD-1 mice. The pulmonary toxicity appears to depend on the metabolism of dauricine mediated by CYP3A. The electrophilic quinone methide metabolite probably plays an important role in the pulmonary toxicity induced by dauricine. PMID- 20008064 TI - Complete genome sequence of the wild-type commensal Escherichia coli strain SE15, belonging to phylogenetic group B2. AB - Escherichia coli SE15 (O150:H5) is a human commensal bacterium recently isolated from feces of a healthy adult and classified into E. coli phylogenetic group B2, which includes the majority of extraintestinal pathogenic E. coli. Here, we report the finished and annotated genome sequence of this organism. PMID- 20008065 TI - ExsD inhibits expression of the Pseudomonas aeruginosa type III secretion system by disrupting ExsA self-association and DNA binding activity. AB - Pseudomonas aeruginosa utilizes a type III secretion system (T3SS) to damage eukaryotic host cells and evade phagocytosis. Transcription of the T3SS regulon is controlled by ExsA, a member of the AraC/XylS family of transcriptional regulators. ExsA-dependent transcription is coupled to type III secretory activity through a cascade of three interacting proteins (ExsC, ExsD, and ExsE). Genetic data suggest that ExsD functions as an antiactivator by preventing ExsA dependent transcription, ExsC functions as an anti-antiactivator by binding to and inhibiting ExsD, and ExsE binds to and inhibits ExsC. T3SS gene expression is activated in response to low-calcium growth conditions or contact with host cells, both of which trigger secretion of ExsE. In the present study we reconstitute the T3SS regulatory cascade in vitro using purified components and find that the ExsD.ExsA complex lacks DNA binding activity. As predicted by the genetic data, ExsC addition dissociates the ExsD.ExsA complex through formation of an ExsD.ExsC complex, thereby releasing ExsA to bind T3SS promoters and activate transcription. Addition of ExsE to the purified system results in formation of the ExsE.ExsC complex and prevents ExsC from dissociating the ExsD.ExsA complex. Although purified ExsA is monomeric in solution, bacterial two hybrid analyses demonstrate that ExsA can self-associate and that ExsD inhibits self-association of ExsA. Based on these data we propose a model in which ExsD regulates ExsA-dependent transcription by inhibiting the DNA-binding and self association properties of ExsA. PMID- 20008066 TI - Identification of conserved amino acid residues of the Salmonella sigmaS chaperone Crl involved in Crl-sigmaS interactions. AB - Proteins that bind sigma factors typically attenuate the function of the sigma factor by restricting its access to the RNA polymerase (RNAP) core enzyme. An exception to this general rule is the Crl protein that binds the stationary-phase sigma factor sigma(S) (RpoS) and enhances its affinity for the RNAP core enzyme, thereby increasing expression of sigma(S)-dependent genes. Analyses of sequenced bacterial genomes revealed that crl is less widespread and less conserved at the sequence level than rpoS. Seventeen residues are conserved in all members of the Crl family. Site-directed mutagenesis of the crl gene from Salmonella enterica serovar Typhimurium and complementation of a Deltacrl mutant of Salmonella indicated that substitution of the conserved residues Y22, F53, W56, and W82 decreased Crl activity. This conclusion was further confirmed by promoter binding and abortive transcription assays. We also used a bacterial two-hybrid system (BACTH) to show that the four substitutions in Crl abolish Crl-sigma(S) interaction and that residues 1 to 71 in sigma(S) are dispensable for Crl binding. In Escherichia coli, it has been reported that Crl also interacts with the ferric uptake regulator Fur and that Fur represses crl transcription. However, the Salmonella Crl and Fur proteins did not interact in the BACTH system. In addition, a fur mutation did not have any significant effect on the expression level of Crl in Salmonella. These results suggest that the relationship between Crl and Fur is different in Salmonella and E. coli. PMID- 20008067 TI - Impact of silver(I) on the metabolism of Shewanella oneidensis. AB - Anaerobic cultures of Shewanella oneidensis MR-1 reduced toxic Ag(I), forming nanoparticles of elemental Ag(0), as confirmed by X-ray diffraction analyses. The addition of 1 to 50 microM Ag(I) had a limited impact on growth, while 100 microM Ag(I) reduced both the doubling time and cell yields. At this higher Ag(I) concentration transmission electron microscopy showed the accumulation of elemental silver particles within the cell, while at lower concentrations the metal was exclusively reduced and precipitated outside the cell wall. Whole organism metabolite fingerprinting, using the method of Fourier transform infrared spectroscopy analysis of cells grown in a range of silver concentrations, confirmed that there were significant physiological changes at 100 microM silver. Principal component-discriminant function analysis scores and loading plots highlighted changes in certain functional groups, notably, lipids, amides I and II, and nucleic acids, as being discriminatory. Molecular analyses confirmed a dramatic drop in cellular yields of both the phospholipid fatty acids and their precursor molecules at high concentrations of silver, suggesting that the structural integrity of the cellular membrane was compromised at high silver concentrations, which was a result of intracellular accumulation of the toxic metal. PMID- 20008068 TI - Extracytoplasmic PAS-like domains are common in signal transduction proteins. AB - We present the crystal structure of the extracytoplasmic domain of the Bacillus subtilis PhoR sensor histidine kinase, part of a two-component system involved in adaptation to low environmental phosphate concentrations. In addition to the PhoR structure, we predict that the majority of the extracytoplasmic domains of B. subtilis sensor kinases will adopt a fold similar to the ubiquitous PAS domain. PMID- 20008069 TI - Virulence factors encoded by Legionella longbeachae identified on the basis of the genome sequence analysis of clinical isolate D-4968. AB - Legionella longbeachae causes most cases of legionellosis in Australia and may be underreported worldwide due to the lack of L. longbeachae-specific diagnostic tests. L. longbeachae displays distinctive differences in intracellular trafficking, caspase 1 activation, and infection in mouse models compared to Legionella pneumophila, yet these two species have indistinguishable clinical presentations in humans. Unlike other legionellae, which inhabit freshwater systems, L. longbeachae is found predominantly in moist soil. In this study, we sequenced and annotated the genome of an L. longbeachae clinical isolate from Oregon, isolate D-4968, and compared it to the previously published genomes of L. pneumophila. The results revealed that the D-4968 genome is larger than the L. pneumophila genome and has a gene order that is different from that of the L. pneumophila genome. Genes encoding structural components of type II, type IV Lvh, and type IV Icm/Dot secretion systems are conserved. In contrast, only 42/140 homologs of genes encoding L. pneumophila Icm/Dot substrates have been found in the D-4968 genome. L. longbeachae encodes numerous proteins with eukaryotic motifs and eukaryote-like proteins unique to this species, including 16 ankyrin repeat-containing proteins and a novel U-box protein. We predict that these proteins are secreted by the L. longbeachae Icm/Dot secretion system. In contrast to the L. pneumophila genome, the L. longbeachae D-4968 genome does not contain flagellar biosynthesis genes, yet it contains a chemotaxis operon. The lack of a flagellum explains the failure of L. longbeachae to activate caspase 1 and trigger pyroptosis in murine macrophages. These unique features of L. longbeachae may reflect adaptation of this species to life in soil. PMID- 20008070 TI - The cyclic nucleotide monophosphate domain of Xanthomonas campestris global regulator Clp defines a new class of cyclic di-GMP effectors. AB - The widely conserved second messenger cyclic diguanosine monophosphate (c-di-GMP) plays a key role in quorum-sensing (QS)-dependent production of virulence factors in Xanthomonas campestris pv. campestris. The detection of QS diffusible signal factor (DSF) by the sensor RpfC leads to the activation of response regulator RpfG, which activates virulence gene expression by degrading c-di-GMP. Here, we show that a global regulator in the X. campestris pv. campestris QS regulatory pathway, Clp, is a c-di-GMP effector. c-di-GMP specifically binds to Clp with high affinity and induces allosteric conformational changes that abolish the interaction between Clp and its target gene promoter. Clp is similar to the cyclic AMP (cAMP) binding proteins Crp and Vfr and contains a conserved cyclic nucleotide monophosphate (cNMP) binding domain. Using site-directed mutagenesis, we found that the cNMP binding domain of Clp contains a glutamic acid residue (E99) that is essential for c-di-GMP binding. Substituting the residue with serine (E99S) resulted in decreased sensitivity to changes in the intracellular c di-GMP level and attenuated bacterial virulence. These data establish the direct role of Clp in the response to fluctuating c-di-GMP levels and depict a novel mechanism by which QS links the second messenger with the X. campestris pv. campestris virulence regulon. PMID- 20008071 TI - Protein localization in Escherichia coli cells: comparison of the cytoplasmic membrane proteins ProP, LacY, ProW, AqpZ, MscS, and MscL. AB - Fluorescence microscopy has revealed that the phospholipid cardiolipin (CL) and FlAsH-labeled transporters ProP and LacY are concentrated at the poles of Escherichia coli cells. The proportion of CL among E. coli phospholipids can be varied in vivo as it is decreased by cls mutations and it increases with the osmolality of the growth medium. In this report we compare the localization of CL, ProP, and LacY with that of other cytoplasmic membrane proteins. The proportion of cells in which FlAsH-labeled membrane proteins were concentrated at the cell poles was determined as a function of protein expression level and CL content. Each tagged protein was expressed from a pBAD24-derived plasmid; tagged ProP was also expressed from the chromosome. The osmosensory transporter ProP and the mechanosensitive channel MscS concentrated at the poles at frequencies correlated with the cellular CL content. The lactose transporter LacY was found at the poles at a high and CL-independent frequency. ProW (a component of the osmoregulatory transporter ProU), AqpZ (an aquaporin), and MscL (a mechanosensitive channel) were concentrated at the poles in a minority of cells, and this polar localization was CL independent. The frequency of polar localization was independent of induction (at arabinose concentrations up to 1 mM) for proteins encoded by pBAD24-derived plasmids. Complementation studies showed that ProW, AqpZ, MscS, and MscL remained functional after introduction of the FlAsH tag (CCPGCC). These data suggest that CL-dependent polar localization in E. coli cells is not a general characteristic of transporters, channels, or osmoregulatory proteins. Polar localization can be frequent and CL independent (as observed for LacY), frequent and CL dependent (as observed for ProP and MscS), or infrequent (as observed for AqpZ, ProW, and MscL). PMID- 20008072 TI - Genetic analysis of the regulation of type IV pilus function by the Chp chemosensory system of Pseudomonas aeruginosa. AB - The virulence of the opportunistic pathogen Pseudomonas aeruginosa involves the coordinate expression of many virulence factors, including type IV pili, which are required for colonization of host tissues and for twitching motility. Type IV pilus function is controlled in part by the Chp chemosensory system, which includes a histidine kinase, ChpA, and two CheY-like response regulators, PilG and PilH. How the Chp components interface with the type IV pilus motor proteins PilB, PilT, and PilU is unknown. We present genetic evidence confirming the role of ChpA, PilG, and PilB in the regulation of pilus extension and the role of PilH and PilT in regulating pilus retraction. Using informative double and triple mutants, we show that (i) ChpA, PilG, and PilB function upstream of PilH, PilT, and PilU; (ii) that PilH enhances PilT function; and (iii) that PilT and PilB retain some activity in the absence of signaling input from components of the Chp system. By site-directed mutagenesis, we demonstrate that the histidine kinase domain of ChpA and the phosphoacceptor sites of both PilG and PilH are required for type IV pilus function, suggesting that they form a phosphorelay system important in the regulation of pilus extension and retraction. Finally, we present evidence suggesting that pilA transcription is regulated by intracellular PilA levels. We show that PilA is a negative regulator of pilA transcription in P. aeruginosa and that the Chp system functionally regulates pilA transcription by controlling PilA import and export. PMID- 20008073 TI - PrhG, a transcriptional regulator responding to growth conditions, is involved in the control of the type III secretion system regulon in Ralstonia solanacearum. AB - The ability of Ralstonia solanacearum to cause disease in plants depends on its type III secretion system (T3SS). The expression of the T3SS and its effector substrates is coordinately controlled by a regulatory cascade, at the bottom of which is HrpB. Transcription of the hrpB gene is activated by a plant-responsive regulator named HrpG, which is a master regulator of a wide array of pathogenicity functions in R. solanacearum. We have identified in the genome of strain GMI1000 a close paralog of hrpG (83% overall similarity at the protein level) that we have named prhG. Despite this high similarity, the expression pattern of prhG is remarkably different from that of hrpG: prhG expression is activated after growth of bacteria in minimal medium but not in the presence of host cells, while hrpG expression is specifically induced in response to plant cell signals. We provide genetic evidence that prhG is a transcriptional regulator that, like hrpG, controls the expression of hrpB and the hrpB-regulated genes under minimal medium conditions. However, the regulatory functions of prhG and hrpG are distinct: prhG has no influence on hrpB expression when the bacteria are in the presence of plant cells, and transcriptomic profiling analysis of a prhG mutant revealed that the PrhG and HrpG regulons have only one pathogenicity target in common, hrpB. Functional complementation experiments indicated that PrhG and HrpG are individually sufficient to activate hrpB expression in minimal medium. Rather surprisingly, a prhG disruption mutant had little impact on pathogenicity, which may indicate that prhG has a minor role in the activation of T3SS genes when R. solanacearum grows parasitically inside the plant. The cross talk between pathogenicity regulatory proteins and environmental signals described here denotes that an intricate network is at the basis of the bacterial disease program. PMID- 20008074 TI - Temporal and spatial regulation of the four transcription start sites of hetR from Anabaena sp. strain PCC 7120. AB - The filamentous cyanobacterium Anabaena sp. strain PCC 7120 forms nitrogen-fixing heterocysts in a periodic pattern in response to combined-nitrogen limitation in the environment. The master regulator of heterocyst differentiation, HetR, is necessary for both pattern formation and commitment of approximately every 10th cell of a filament to differentiation into a heterocyst. In this study, the individual contributions of four transcriptional start points (tsps) in regulation of transcription of hetR were assessed, and the effects of the regulatory genes patS, hetN, and patA on transcription from the tsps were determined. The tsp located at nucleotide -271 relative to the translational start site (-271 tsp) was the most tightly regulated tsp, and fluorescence from a -271 tsp-green fluorescent protein (GFP) reporter fusion was observed initially in groups of two cells and later in single cells arranged in a spatial pattern that mimicked the pattern of heterocysts that emerged. Conversely, the fluorescence from the -184 and -728/-696 tsp-GFP reporter fusions was uniform throughout filaments. Transcription from the -271 tsp was severely downregulated in a strain in which the patA gene, which encodes a positive regulator of differentiation, was deleted, and it was not detectable in strains overexpressing the genes encoding the negative regulators of differentiation, patS and hetN. In strains lacking the -271 tsp of hetR, pattern formation, the timing of commitment to differentiation, and the number of cells that differentiated into heterocysts were affected. Taken together, these results demonstrate the role of regulation of the -271 tsp of hetR in the genetic network that governs heterocyst pattern formation and differentiation. PMID- 20008075 TI - Comparative genomics and transduction potential of Enterococcus faecalis temperate bacteriophages. AB - To determine the relative importance of temperate bacteriophage in the horizontal gene transfer of fitness and virulence determinants of Enterococcus faecalis, a panel of 47 bacteremia isolates were treated with the inducing agents mitomycin C, norfloxacin, and UV radiation. Thirty-four phages were purified from culture supernatants and discriminated using pulsed-field gel electrophoresis (PFGE) and restriction mapping. From these analyses the genomes of eight representative phages were pyrosequenced, revealing four distinct groups of phages. Three groups of phages, PhiFL1 to 3, were found to be sequence related, with PhiFL1A to C and PhiFL2A and B sharing the greatest identity (87 to 88%), while PhiFL3A and B share 37 to 41% identity with PhiFL1 and 2. PhiFL4A shares 3 to 12% identity with the phages PhiFL1 to 3. The PhiFL3A and B phages possess a high DNA sequence identity with the morphogenesis and lysis modules of Lactococcus lactis subsp. cremoris prophages. Homologs of the Streptococcus mitis platelet binding phage tail proteins, PblA and PblB, are encoded on each sequenced E. faecalis phage. Few other phage genes encoding potential virulence functions were identified, and there was little evidence of carriage of lysogenic conversion genes distal to endolysin, as has been observed with genomes of many temperate phages from the opportunist pathogens Staphylococcus aureus and Streptococcus pyogenes. E. faecalis JH2-2 lysogens were generated using the eight phages, and these were examined for their relative fitness in Galleria mellonella. Several lysogens exhibited different effects upon survival of G. mellonella compared to their isogenic parent. The eight phages were tested for their ability to package host DNA, and three were shown to be very effective for generalized transduction of naive host cells of the laboratory strains OG1RF and JH2-2. PMID- 20008076 TI - Adiponectin and cardiac geometry and function in elderly: results from two community-based cohort studies. AB - OBJECTIVE: Several smaller studies have indicated that adiponectin might be associated with left ventricular (LV) mass and function, but community-based studies with adequate sample size and adjustment for potential confounders are lacking. Our objective was to investigate such associations in two large community-based studies of elderly. DESIGN: Cross-sectional. METHODS: We evaluated cross-sectional relations between serum adiponectin and echocardiographic measures of cardiac geometry and function (LV mass index, LV relative wall thickness, LV end-diastolic diameter, left atrial diameter, ejection fraction, LV isovolumic relaxation time, and E/A ratio) in 954 70-year old participants (50% women) of the Prospective Investigation of the Vasculature in Uppsala Seniors (PIVUS), and in 427 71-year-old men from the Uppsala Longitudinal Study of Adult Men (ULSAM). RESULTS: In models adjusted for age, sex, body mass index, systolic blood pressure, antihypertensive treatment, antidiabetic treatment, lipid-lowering medication, fasting blood glucose, total cholesterol, high-density lipoprotein cholesterol, creatinine, and smoking, adiponectin was inversely associated with ejection fraction in men (beta, -1.62; 95% confidence interval (CI), -2.50, -0.75 in PIVUS; beta, -1.35; 95% CI, -2.41, 0.29 in ULSAM), but not in women. After additional adjustment for N-terminal pro brain natriuretic peptide (NT-proBNP), the association between adiponectin and ejection fraction was attenuated (beta, -0.98; 95% CI, -1.86, -0.10 in PIVUS; beta, -0.75; 95% CI, -1.84, 0.35 in ULSAM). CONCLUSIONS: Serum adiponectin concentrations were associated with ejection fraction in men, and these associations were partially attenuated by NT-proBNP. Our results imply that adiponectin may be associated with systolic function through pathways that involve natriuretic peptides. PMID- 20008077 TI - Polysaccharide of radix pseudostellariae improves chronic fatigue syndrome induced by poly I:C in mice. AB - Radix Pseudostellariae is used as a tonic drug in traditional Chinese medicine with immunomodulating and anti-fatigue activities, and the polysaccharide is considered as the main active component. The purpose of this study is to examine the effect of the polysaccharide isolated from Radix Pseudostellariae (PRP) on mouse chronic fatigue syndrome (CFS) induced by intraperitoneal injection of polyriboinosinic:polyribocytidylic acid (poly I:C), a double-stranded synthetic RNA. It has shown that the fatigue symptom of mice lasted at least 1 week as evaluated by forced swimming time. PRP (100, 200, 400 mg kg(-1)), orally administered 3 days before poly I:C injection, showed dose-dependent anti-fatigue effects. In addition, poly I:C led to evident alternations in neuroendocrine and immune systems of mice, such as reduced spontaneous activity and learning ability, declined serum level of corticosterone, increased weight indexes and T lymphocyte numbers in thymuses and spleens, and increased CD4(+)/CD8(+) ratio but decreased proliferation ability of T lymphocytes in spleens. PRP alleviated the abnormalities caused by poly I:C, and restored the function of hosts to normal conditions. The findings suggest that PRP is beneficial to CFS, and the underlying mechanisms of action involve neuroendocrine and immune systems. PMID- 20008078 TI - REC-2006-A Fractionated Extract of Podophyllum hexandrum Protects Cellular DNA from Radiation-Induced Damage by Reducing the Initial Damage and Enhancing Its Repair In Vivo. AB - Podophyllum hexandrum, a perennial herb commonly known as the Himalayan May Apple, is well known in Indian and Chinese traditional systems of medicine. P. hexandrum has been widely used for the treatment of venereal warts, skin infections, bacterial and viral infections, and different cancers of the brain, lung and bladder. This study aimed at elucidating the effect of REC-2006, a bioactive fractionated extract from the rhizome of P. hexandrum, on the kinetics of induction and repair of radiation-induced DNA damage in murine thymocytes in vivo. We evaluated its effect on non-specific radiation-induced DNA damage by the alkaline halo assay in terms of relative nuclear spreading factor (RNSF) and gene specific radiation-induced DNA damage via semi-quantitative polymerase chain reaction. Whole body exposure of animals with gamma rays (10 Gy) caused a significant amount of DNA damage in thymocytes (RNSF values 17.7 +/- 0.47, 12.96 +/- 1.64 and 3.3 +/- 0.014) and a reduction in the amplification of beta-globin gene to 0, 28 and 43% at 0, 15 and 60 min, respectively. Administrating REC-2006 at a radioprotective concentration (15 mg kg(-1) body weight) 1 h before irradiation resulted in time-dependent reduction of DNA damage evident as a decrease in RNSF values 6.156 +/- 0.576, 1.647 +/- 0.534 and 0.496 +/- 0.012, and an increase in beta-globin gene amplification 36, 95 and 99%, at 0, 15 and 60 min, respectively. REC-2006 scavenged radiation-induced hydroxyl radicals in a dose-dependent manner stabilized DPPH free radicals and also inhibited superoxide anions. Various polyphenols and flavonoides present in REC-2006 might contribute to scavenging of radiation-induced free radicals, thereby preventing DNA damage and stimulating its repair. PMID- 20008079 TI - Iron regulation through the back door: iron-dependent metabolite levels contribute to transcriptional adaptation to iron deprivation in Saccharomyces cerevisiae. AB - Budding yeast (Saccharomyces cerevisiae) responds to iron deprivation both by Aft1-Aft2-dependent transcriptional activation of genes involved in cellular iron uptake and by Cth1-Cth2-specific degradation of certain mRNAs coding for iron dependent biosynthetic components. Here, we provide evidence for a novel principle of iron-responsive gene expression. This regulatory mechanism is based on the modulation of transcription through the iron-dependent variation of levels of regulatory metabolites. As an example, the LEU1 gene of branched-chain amino acid biosynthesis is downregulated under iron-limiting conditions through depletion of the metabolic intermediate alpha-isopropylmalate, which functions as a key transcriptional coactivator of the Leu3 transcription factor. Synthesis of alpha-isopropylmalate involves the iron-sulfur protein Ilv3, which is inactivated under iron deficiency. As another example, decreased mRNA levels of the cytochrome c-encoding CYC1 gene under iron-limiting conditions involve heme dependent transcriptional regulation via the Hap1 transcription factor. Synthesis of the iron-containing heme is directly correlated with iron availability. Thus, the iron-responsive expression of genes that are downregulated under iron limiting conditions is conferred by two independent regulatory mechanisms: transcriptional regulation through iron-responsive metabolites and posttranscriptional mRNA degradation. Only the combination of the two processes provides a quantitative description of the response to iron deprivation in yeast. PMID- 20008080 TI - Defects in DNA lesion bypass lead to spontaneous chromosomal rearrangements and increased cell death. AB - Rev3 polymerase and Mph1 DNA helicase participate in error-prone and error-free pathways, respectively, for the bypassing of template lesions during DNA replication. Here we have investigated the role of these pathways and their genetic interaction with recombination factors, other nonreplicative DNA helicases, and DNA damage checkpoint components in the maintenance of genome stability, viability, and sensitivity to the DNA-damaging agent methyl methanesulfonate (MMS). We find that cells lacking Rev3 and Mph1 exhibit a synergistic, Srs2-dependent increase in the rate of accumulating spontaneous, gross chromosomal rearrangements, suggesting that the suppression of point mutations by deletion of REV3 may lead to chromosomal rearrangements. While mph1Delta is epistatic to homologous recombination (HR) genes, both Rad51 and Rad52, but not Rad59, are required for normal growth of the rev3Delta mutant and are essential for survival of rev3Delta cells during exposure to MMS, indicating that Mph1 acts in a Rad51-dependent, Rad59-independent subpathway of HR-mediated lesion bypass. Deletion of MPH1 helicase leads to synergistic DNA damage sensitivity increases in cells with chl1Delta or rrm3Delta helicase mutations, whereas mph1Delta is hypostatic to sgs1Delta. Previously reported slow growth of mph1Delta srs2Delta cells is accompanied by G(2)/M arrest and fully suppressed by disruption of the Mec3-dependent DNA damage checkpoint. We propose a model for replication fork rescue mediated by translesion DNA synthesis and homologous recombination that integrates the role of Mph1 in unwinding D loops and its genetic interaction with Rev3 and Srs2-regulated pathways in the suppression of spontaneous genome rearrangements and in mutation avoidance. PMID- 20008081 TI - GPR11, a putative seven-transmembrane G protein-coupled receptor, controls zoospore development and virulence of Phytophthora sojae. AB - G protein-coupled receptors (GPCRs) represent a large receptor family involved in a broad spectrum of cell signaling. To understand signaling mechanisms mediated by GPCRs in Phytophthora sojae, we identified and characterized the PsGPR11 gene, which encodes a putative seven-transmembrane GPCR. An expression analysis revealed that PsGPR11 was differentially expressed during asexual development. The highest expression level occurred in zoospores and was upregulated during early infection. PsGPR11-deficienct transformants were obtained by gene silencing strategies. Silenced transformants exhibited no differences in hyphal growth or morphology, sporangium production or size, or mating behavior. However, the release of zoospores from sporangia was severely impaired in the silenced transformants, and about 50% of the sporangia did not completely release their zoospores. Zoospore encystment and germination were also impaired, and zoospores of the transformants lost their pathogenicity to soybean. In addition, no interaction was observed between PsGPR11 and PsGPA1 with a conventional yeast two hybrid assay, and the transcriptional levels of some genes which were identified as being negatively regulated by PsGPA1 were not clearly altered in PsGPR11 silenced mutants. These results suggest that PsGPR11-mediated signaling controls P. sojae zoospore development and virulence through the pathways independent of G protein. PMID- 20008082 TI - WD repeat domain of Dictyostelium myosin heavy chain kinase C functions in both substrate targeting and cellular localization. AB - Myosin II disassembly in Dictyostelium discoideum is regulated by three structurally related myosin heavy chain kinases (myosin II heavy chain kinase A [MHCK-A], -B, and -C). We show that the WD repeat domain of MHCK-C is unique in that it mediates both substrate targeting and subcellular localization, revealing a target for regulation that is distinct from those of the other MHCKs. PMID- 20008083 TI - Impaired autoregulation of cerebral blood flow during rewarming from hypothermic cardiopulmonary bypass and its potential association with stroke. AB - BACKGROUND: Patient rewarming after hypothermic cardiopulmonary bypass (CPB) has been linked to brain injury after cardiac surgery. In this study, we evaluated whether cooling and then rewarming of body temperature during CPB in adult patients is associated with alterations in cerebral blood flow (CBF)-blood pressure autoregulation. METHODS: One hundred twenty-seven adult patients undergoing CPB during cardiac surgery had transcranial Doppler monitoring of the right and left middle cerebral artery blood flow velocity. Eleven patients undergoing CPB who had arterial inflow maintained at >35 degrees C served as controls. The mean velocity index (Mx) was calculated as a moving, linear correlation coefficient between slow waves of middle cerebral artery blood flow velocity and mean arterial blood pressure. Intact CBF-blood pressure autoregulation is associated with an Mx that approaches 0. Impaired autoregulation results in an increasing Mx approaching 1.0. Comparisons of time averaged Mx values were made between the following periods: before CPB (baseline), during the cooling and rewarming phases of CPB, and after CPB. The number of patients in each phase of CPB with an Mx >4.0, indicative of impaired CBF autoregulation, was determined. RESULTS: During cooling, Mx (left, 0.29 +/- 0.18; right, 0.28 +/- 0.18 [mean +/- SD]) was greater than that at baseline (left, 0.17 +/- 0.21; right, 0.17 +/- 0.20; P or=0.4 during the cooling phase of CPB and 68 (53%) had an average Mx >or=0.4 during rewarming. Nine of the 11 warm controls had an average Mx >or=0.4 during the entire CPB period. There were 7 strokes and 1 TIA after surgery. All strokes were in patients with Mx >or= 0.4 during rewarming (P = 0.015). The unadjusted odds ratio for any neurologic event (stroke or transient ischemic attack) for patients with Mx >or= 0.4 during rewarming was 6.57 (95% confidence interval, 0.79 to 55.0, P < 0.08). CONCLUSIONS: Hypothermic CPB is associated with abnormal CBF blood pressure autoregulation that is worsened with rewarming. We found a high rate of strokes in patients with evidence of impaired CBF autoregulation. Whether a pressure-passive CBF state during rewarming is associated with risk for ischemic brain injury requires further investigation. PMID- 20008085 TI - Ectopic bone formation from mandibular osteoblasts cultured in a novel human serum-derived albumin scaffold. AB - The aim of this study was to evaluate the ectopic bone formation using a novel serum-derived albumin scaffold and cultured human mandibular osteoblasts in nude mice. Osteoblasts were cultured with 10% human serum and plated in a novel spongy noncalcified protein scaffold prepared with plasmatic albumin crossed with a glutaraldehyde type agent. Hematoxylin-eosin staining revealed a bone-like extracellular matrix and in vitro mineralization was confirmed by von Kossa staining. Histological and immunohistochemical evaluation showed progression of mineralization in vivo. These results suggest the clinical feasibility of alveolar cells and albumin scaffold as a good alternative for bone regeneration. PMID- 20008084 TI - Demographic, maltreatment, and neurobiological correlates of PTSD symptoms in children and adolescents. AB - OBJECTIVE: To examine the relationships of demographic, maltreatment, neurostructural and neuropsychological measures with total posttraumatic stress disorder (PTSD) symptoms. METHODS: Participants included 216 children with maltreatment histories (N = 49), maltreatment and PTSD (N = 49), or no maltreatment (N = 118). Participants received diagnostic interviews, brain imaging, and neuropsychological evaluations. RESULTS: We examined a hierarchical regression model comprised of independent variables including demographics, trauma and maltreatment-related variables, and hippocampal volumes and neuropsychological measures to model PTSD symptoms. Important independent contributors to this model were SES, and General Maltreatment and Sexual Abuse Factors. Although hippocampal volumes were not significant, Visual Memory was a significant contributor to this model. CONCLUSIONS: Similar to adult PTSD, pediatric PTSD symptoms are associated with lower Visual Memory performance. It is an important correlate of PTSD beyond established predictors of PTSD symptoms. These results support models of developmental traumatology and suggest that treatments which enhance visual memory may decrease symptoms of PTSD. PMID- 20008086 TI - Oral keratinocyte responses to nickel-based dental casting alloys in vitro. AB - Adverse reactions of oral mucosa to nickel-based dental casting alloys are probably due to corrosion metal ion release. We exposed H400 oral keratinocytes to two Ni-based dental alloys (Matchmate and Dsign10) as well as NiCl( 2) (1-40 microg/mL Ni(2+)). Alloy derived Ni(2+) media concentrations were determined. Direct culture on both alloys resulted in inhibited growth with a greater effect observed for Dsign10 (higher ion release). Indirect exposure of cells to conditioned media from Dsign10 negatively affected cell numbers (approximately 64% of control by 6 days) and morphology while Matchmate-derived media did not. Exposure to increasing NiCl(2) negatively affected cell growth and morphology, and the Granulocyte-macrophage colony-stimulating factor (GM-CSF) transcript was significantly up-regulated in cells following direct and indirect exposure to Dsign10. NiCl(2) exposure up-regulated all cytokine transcripts at 1 day. At day 6, IL-1beta and IL-8 transcripts were suppressed while GM-CSF and IL-11 increased with Ni(2+) dose. Accumulation of Ni(2+) ions from alloys in oral tissues may affect keratinocyte viability and chronic inflammation. PMID- 20008087 TI - Antimicrobial properties of light-activated polyurethane containing indocyanine green. AB - The aim of this study was to produce novel antimicrobial polymers containing the light-activated antimicrobial agent indocyanine green (ICG). The novel materials were prepared by swelling polyurethane in acetone containing water and ICG, followed by solvent evaporation. The uptake of ICG was dependent upon the ratio of acetone to water. Only at a ratio of 99 parts acetone to 1 part water was there any substantial colouration of the samples. When exposed to laser light from the near infrared spectrum (808 nm), polyurethane-containing ICG exhibited antimicrobial activity against Gram-positive bacteria; a 2 log10 reduction was achieved against methicillin-resistant Staphylococcus aureus (MRSA) and Staphylococcus epidermidis after 15 min exposure, corresponding to an energy dose of 31.83 J delivered at an energy density of 31.83 J/cm2. Under the same conditions, Gram-negative bacteria (Escherichia coli and Pseudomonas aeruginosa) appeared to be less susceptible, the viable count being reduced by 0.5 log10. Some of the physical properties of the resulting material were also investigated and it was found that the elasticity (Young's modulus) was reduced by approximately 60%. Furthermore, when the ICG-containing polymer was stretched, the breaking point occurred when the elongation was 6.7 times the initial value, while ICG-free polyurethane samples did not break following a 7-fold elongation. The contact angles for water droplets revealed that the ICG-containing polymer was more hydrophobic than untreated polyurethane. The results of this study show that ICG can be embedded in polyurethane to produce materials which when irradiated with near-infrared light can exert a bactericidal effect particularly against MRSA and S. epidermidis. Such materials may be useful for preparing intravenous catheters, which are often colonized by such organisms. PMID- 20008088 TI - The response of macrophages to a Cu-Al-Ni shape memory alloy. AB - Cu-Al-Ni shape memory alloys (SMAs) have been investigated as materials for medical devices, but little is known about their biocompatibility. The aim of this work was to study the response of rat peritoneal macrophages (PMO) to a Cu Al-Ni SMA in vitro, by measuring the functional activity of mitochondria, necrosis, apoptosis, and production of proinflammatory cytokines. Rapidly solidified (RS) thin ribbons were used for the tests. The control alloy was a permanent mold casting of the same composition, but without the shape memory effect. Our results showed that the control alloy was severely cytotoxic, whereas RS ribbons induced neither necrosis nor apoptosis of PMO. These findings correlated with the data that RS ribbons are significantly more resistant to corrosion compared to the control alloy, as judged by the lesser release of Cu and Ni in the conditioning medium. However, the ribbons generated intracellular reactive oxygen species and upregulated the production of IL-6 by PMO. These effects were almost completely abolished by conditioning the RS ribbons for 5 weeks. In conclusion, RS significantly improves the corrosion stability and biocompatibility of Cu-Al-Ni SMA. The biocompatibility of this functional material could be additionally enhanced by conditioning the ribbons in cell culture medium. PMID- 20008089 TI - The axillary artery as an alternative site of cannulation for redo port access assisted minimally invasive mitral valve surgery: early report of 2 cases. AB - The minimally invasive Heartport (HP)-assisted technique has become first choice option for mitral valve surgery in many centres.The pool of patients potentially treated using HP techniques, however, is still limited by the presence of peripheral vessel disease, expecially in the elderly population. Alternative approaches to using the HP technique safely in such a subset of patients, therefore, should be evaluated. Here, we present our preliminary experience using the axillary artery as an alternative site of cannulation for HP-assisted redo mitral valve surgery in patients with concomitant peripheral vessel disease. PMID- 20008090 TI - The PTPN22 R620W polymorphism is not associated with systemic rheumatic diseases in South Africans. PMID- 20008091 TI - Human leukocyte antigen-G in ankylosing spondylitis and the response after tumour necrosis factor-alpha blocker therapy. AB - OBJECTIVE: To investigate the role of HLA-G in AS. METHODS: Serum levels of soluble HLA-G (sHLA-G) were measured in 80 AS patients and 30 healthy controls. The expression of HLA-G on the peripheral blood mononuclear cell (PBMC) surface was investigated in the same 80 AS patients and 40 healthy controls by flow cytometry. The response of HLA-G after 3 months of TNF-alpha blocker therapy (adalimumab) was evaluated in 14 AS patients. We evaluated Bath Ankylosing Spondylitis Disease Activity Index (BASDAI), Bath Ankylosing Spondylitis Functional Index (BASFI), Bath Ankylosing Spondylitis Patient Global Score (BAS G), physical mobility, ESR and CRP levels. RESULTS: Serum levels of sHLA-G were significantly lower in 80 AS patients than 30 healthy controls [mean (s.d.) 22.47 (26.8) vs 34.78 (32.01) U/ml, P = 0.028], and correlated significantly with modified Schober index (r = 0.326; P = 0.009), chest expansion (r = 0.319; P = 0.011), lateral lumbar flexion (r = 0.377; P = 0.002), cervical rotation (r = 0.396; P = 0.004), whereas inversely correlated with fingertip-to-floor distance (r = -0.282; P = 0.026) and tragus-to-wall distance (r = -0.270; P = 0.031). The expression of HLA-G on PBMCs was significantly higher in 80 AS patients than 40 healthy controls [mean (s.d.) 18.5 (6.10)% vs 15.41 (4.84)%; P = 0.012], and correlated significantly with ESR (r = 0.421; P < 0.001) and CRP (r = 0.419; P < 0.001). The expression of HLA-G on PMBCs decreased significantly after 3 months of adalimumab therapy [third month vs baseline, 13.46 (5.38)% vs 19.87 (7.31)%; P = 0.016]. CONCLUSIONS: Lower serum levels of sHLA-G contribute to susceptibility to AS, and predispose to poor spinal mobility. The expression of HLA-G on PMBCs is up-regulated in AS, correlates with acute phase reactants and decreases after TNF-alpha blocker therapy, suggesting an index of disease activity. PMID- 20008092 TI - Treatment of a patient with remitting seronegative, symmetrical synovitis with pitting oedema with a humanized anti-interleukin-6 receptor antibody, tocilizumab. PMID- 20008093 TI - First-line DMARD choice in early rheumatoid arthritis--do prognostic factors play a role? AB - OBJECTIVE: To examine if prognostic factors predict the choice of first DMARD for patients with RA. METHODS: Details of 616 patients with early RA were collected from 16 centres in the UK Early Rheumatoid Arthritis Network (ERAN). Logistic regression was used to identify whether HAQ score, swollen joint count (SJC), nodules, RF, ESR, CRP and erosions on radiographs were associated with the choice of first DMARD treatment. RESULTS: Of 616 patients, 547 (88%) were started on a DMARD, 253 (46%) on MTX, 230 (42%) on SSZ, 47 (9%) on other DMARD monotherapies and 17 (3%) on combination DMARD therapy (CoT). SSZ was started less frequently in patients with positive RF (P = 0.018; OR 0.59; 95% CI 0.38, 0.91) and high SJC (P = 0.02; OR 0.95; 95% CI 0.91, 0.99). MTX was favoured in patients with high SJC (P = 0.002; OR 1.07; 95% CI 1.02, 1.11). Non-prescription of DMARDs was associated with old age (P = 0.02; OR 0.98; 95% CI 0.96, 0.99) and low HAQ score (P = 0.009; OR 0.80; 95% CI 0.68, 0.95). None of the variables predicted CoT. All other variables and the hospital where the patient was treated were not independently associated with the choice of DMARD. CONCLUSIONS: When choosing DMARD monotherapy in early RA, rheumatologists in ERAN seem to preferentially prescribe MTX for patients with a poor prognosis and SSZ for patients with good prognosis. No DMARDs were used in older patients or in those with a low HAQ. PMID- 20008096 TI - Dynamic landscapes of four histone modifications during deetiolation in Arabidopsis. AB - Although landscapes of several histone marks are now available for Arabidopsis thaliana and Oryza sativa, such profiles remain static and do not provide information about dynamic changes of plant epigenomes in response to developmental or environmental cues. Here, we analyzed the effects of light on four histone modifications (acetylation and trimethylation of lysines 9 and 27 on histone H3: H3K9ac, H3K9me3, H3K27ac, and H3K27me3, respectively). Our genome wide profiling of H3K9ac and H3K27ac revealed that these modifications are nontransposable element gene-specific. By contrast, we found that H3K9me3 and H3K27me3 target nontransposable element genes, but also intergenic regions and transposable elements. Specific light conditions affected the number of modified regions as well as the overall correlation strength between the presence of specific modifications and transcription. Furthermore, we observed that acetylation marks not only ELONGATED HYPOCOTYL5 and HY5-HOMOLOG upon deetiolation, but also their downstream targets. We found that the activation of photosynthetic genes correlates with dynamic acetylation changes in response to light, while H3K27ac and H3K27me3 potentially contribute to light regulation of the gibberellin metabolism. Thus, this work provides a dynamic portrait of the variations in histone modifications in response to the plant's changing light environment and strengthens the concept that histone modifications represent an additional layer of control for light-regulated genes involved in photomorphogenesis. PMID- 20008097 TI - Systemic therapy for neuroendocrine tumours of gastroenteropancreatic origin. AB - Systemic therapy is one of a number of treatment options routinely used in the management of advanced, unresectable neuroendocrine tumours (NETs). In contrast to many of the other NET treatment modalities, there is at least some evidence base to justify its use. Even so, well-designed clinical trials are limited, since conducting clinical research in this complex group of rare cancers is challenging. The remit of this review article is to summarise the oncology literature and explain the role of systemic therapy in treating NETs of gastroenteropancreatic origin, identifying benefits and limitations. The molecular biology of NETs is now being unravelled, which affords new opportunities for development of mechanism-driven therapies. The rationale for some of the newer systemic targeted therapies that are showing promise in the clinic is discussed. PMID- 20008098 TI - Leptin-R and its association with PI3K/AKT signaling pathway in papillary thyroid carcinoma. AB - The putative role of leptin and its receptor (Ob-R) in the pathogenesis of various primary human malignancies has been reported; however, their role in papillary thyroid cancer (PTC) has not yet been evaluated. We investigated the role of Ob-R in a large tissue microarray cohort of PTC followed by in vitro studies using a panel of PTC cell lines. Ob-R overexpression was seen in 80% PTCs and was significantly associated with poor disease-free survival (P=0.0235). PTCs that overexpressed Ob-R showed a aggressive phenotype characterized by older age, extrathyroid extension, larger tumor size, nodal metastasis, advanced stage, tall cell variant histological subtype, and a poor disease-free survival (P=0.0005, P=0.0006, P=0.0398, P=0.0004, P=0.0111, P=0.0003, and P=0.0235 respectively). However, Ob-R expression was not an independent prognostic marker to predict disease-free survival in multivariate analysis. PTCs with overexpression of Ob-R showed a significant direct association with overexpression of XIAP (P<0.0001) and Bcl-XL (P<0.0001). In vitro analysis showed that leptin stimulated cell proliferation and inhibited apoptosis via activation of phosphatidylinisitol 3' kinase (PI3K)/protein kinase B (AKT) signaling pathway. Inhibition of PI3K activity by its inhibitor LY294002 abrogated leptin-mediated PI3K/AKT signaling. Gene silencing of Ob-R in PTC cells resulted in downregulation of phospho-AKT, Bcl-XL, and XIAP expression suggesting that leptin-mediated pathogenesis of PTC occurs via involvement of these downstream targets. Altogether, these data show that leptin plays an important role in PTC pathogenesis through PI3K/AKT pathway via Ob-R and is a potential prognostic marker associated with an aggressive phenotype and poor disease-free survival. PMID- 20008099 TI - Quantitation of cellular deoxynucleoside triphosphates. AB - Eukaryotic cells contain a delicate balance of minute amounts of the four deoxyribonucleoside triphosphates (dNTPs), sufficient only for a few minutes of DNA replication. Both a deficiency and a surplus of a single dNTP may result in increased mutation rates, faulty DNA repair or mitochondrial DNA depletion. dNTPs are usually quantified by an enzymatic assay in which incorporation of radioactive dATP (or radioactive dTTP in the assay for dATP) into specific synthetic oligonucleotides by a DNA polymerase is proportional to the concentration of the unknown dNTP. We find that the commonly used Klenow DNA polymerase may substitute the corresponding ribonucleotide for the unknown dNTP leading in some instances to a large overestimation of dNTPs. We now describe assay conditions for each dNTP that avoid ribonucleotide incorporation. For the dTTP and dATP assays it suffices to minimize the concentrations of the Klenow enzyme and of labeled dATP (or dTTP); for dCTP and dGTP we had to replace the Klenow enzyme with either the Taq DNA polymerase or Thermo Sequenase. We suggest that in some earlier reports ribonucleotide incorporation may have caused too high values for dGTP and dCTP. PMID- 20008100 TI - SeqBuster, a bioinformatic tool for the processing and analysis of small RNAs datasets, reveals ubiquitous miRNA modifications in human embryonic cells. AB - High-throughput sequencing technologies enable direct approaches to catalog and analyze snapshots of the total small RNA content of living cells. Characterization of high-throughput sequencing data requires bioinformatic tools offering a wide perspective of the small RNA transcriptome. Here we present SeqBuster, a highly versatile and reliable web-based toolkit to process and analyze large-scale small RNA datasets. The high flexibility of this tool is illustrated by the multiple choices offered in the pre-analysis for mapping purposes and in the different analysis modules for data manipulation. To overcome the storage capacity limitations of the web-based tool, SeqBuster offers a stand alone version that permits the annotation against any custom database. SeqBuster integrates multiple analyses modules in a unique platform and constitutes the first bioinformatic tool offering a deep characterization of miRNA variants (isomiRs). The application of SeqBuster to small-RNA datasets of human embryonic stem cells revealed that most miRNAs present different types of isomiRs, some of them being associated to stem cell differentiation. The exhaustive description of the isomiRs provided by SeqBuster could help to identify miRNA-variants that are relevant in physiological and pathological processes. SeqBuster is available at http://estivill_lab.crg.es/seqbuster. PMID- 20008101 TI - Cancer-selective antiproliferative activity is a general property of some G-rich oligodeoxynucleotides. AB - Oligodeoxynucleotide libraries containing randomly incorporated bases are used to generate DNA aptamers by systematic evolution of ligands by exponential enrichment (SELEX). We predicted that combinatorial libraries with alternative base compositions might have innate properties different from the standard library containing equimolar A + C + G + T bases. In particular, we hypothesized that G-rich libraries would contain a higher proportion of quadruplex-forming sequences, which may impart desirable qualities, such as increased nuclease resistance and enhanced cellular uptake. Here, we report on 11 synthetic oligodeoxynucleotide libraries of various base combinations and lengths, with regard to their circular dichroism, stability in serum-containing medium, cellular uptake, protein binding and antiproliferative activity. Unexpectedly, we found that some G-rich libraries (composed of G + T or G + C nucleotides) strongly inhibited cancer cell growth while sparing non-malignant cells. These libraries had spectral features consistent with G-quadruplex formation, were significantly more stable in serum than inactive libraries and showed enhanced cellular uptake. Active libraries generally had strong protein binding, while the pattern of protein binding suggested that G/T and G/C libraries have distinct mechanisms of action. In conclusion, cancer-selective antiproliferative activity may be a general feature of certain G-rich oligodeoxynucleotides and is associated with quadruplex formation, nuclease resistance, efficient cellular uptake and protein binding. PMID- 20008102 TI - PCRPi: Presaging Critical Residues in Protein interfaces, a new computational tool to chart hot spots in protein interfaces. AB - Protein-protein interactions (PPIs) are ubiquitous in Biology, and thus offer an enormous potential for the discovery of novel therapeutics. Although protein interfaces are large and lack defining physiochemical traits, is well established that only a small portion of interface residues, the so-called hot spot residues, contribute the most to the binding energy of the protein complex. Moreover, recent successes in development of novel drugs aimed at disrupting PPIs rely on targeting such residues. Experimental methods for describing critical residues are lengthy and costly; therefore, there is a need for computational tools that can complement experimental efforts. Here, we describe a new computational approach to predict hot spot residues in protein interfaces. The method, called Presaging Critical Residues in Protein interfaces (PCRPi), depends on the integration of diverse metrics into a unique probabilistic measure by using Bayesian Networks. We have benchmarked our method using a large set of experimentally verified hot spot residues and on a blind prediction on the protein complex formed by HRAS protein and a single domain antibody. Under both scenarios, PCRPi delivered consistent and accurate predictions. Finally, PCRPi is able to handle cases where some of the input data is either missing or not reliable (e.g. evolutionary information). PMID- 20008103 TI - PCNA and XPF cooperate to distort DNA substrates. AB - XPF is a structure-specific endonuclease that preferentially cleaves 3' DNA flaps during a variety of repair processes. The crystal structure of a crenarchaeal XPF protein bound to a DNA duplex yielded insights into how XPF might recognise branched DNA structures, and recent kinetic data have demonstrated that the sliding clamp PCNA acts as an essential cofactor, possibly by allowing XPF to distort the DNA structure into a proper conformation for efficient cleavage to occur. Here, we investigate the solution structure of the 3'-flap substrate bound to XPF in the presence and absence of PCNA using intramolecular Forster resonance energy transfer (FRET). We demonstrate that recognition of the flap substrate by XPF involves major conformational changes of the DNA, including a 90 degrees kink of the DNA duplex and organization of the single-stranded flap. In the presence of PCNA, there is a further substantial reorganization of the flap substrate bound to XPF, providing a structural basis for the observation that PCNA has an essential catalytic role in this system. The wider implications of these observations for the plethora of PCNA-dependent enzymes are discussed. PMID- 20008104 TI - H2A.Bbd: an X-chromosome-encoded histone involved in mammalian spermiogenesis. AB - Despite the identification of H2A.Bbd as a new vertebrate-specific replacement histone variant several years ago, and despite the many in vitro structural characterizations using reconstituted chromatin complexes consisting of this variant, the existence of H2A.Bbd in the cell and its location has remained elusive. Here, we report that the native form of this variant is present in highly advanced spermiogenic fractions of mammalian testis at the time when histones are highly acetylated and being replaced by protamines. It is also present in the nucleosomal chromatin fraction of mature human sperm. The ectopically expressed non-tagged version of the protein is associated with micrococcal nuclease-refractory insoluble fractions of chromatin and in mouse (20T1/2) cell line, H2A.Bbd is enriched at the periphery of chromocenters. The exceedingly rapid evolution of this unique X-chromosome-linked histone variant is shared with other reproductive proteins including those associated with chromatin in the mature sperm (protamines) of many vertebrates. This common rate of evolution provides further support for the functional and structural involvement of this protein in male gametogenesis in mammals. PMID- 20008105 TI - Legionella dresdenensis sp. nov., isolated from river water. AB - Legionella-like isolates, strains W03-356(T), W03-357 and W03-359, from three independent water samples from the river Elbe, Germany, were analysed by using a polyphasic approach. Morphological and biochemical characterization revealed that they were Gram-negative, aerobic, non-spore-forming bacilli with a cut glass colony appearance that grew only on L-cysteine-supplemented buffered charcoal yeast extract agar. Phylogenetic analysis based on sequence comparisons of the 16S rRNA, macrophage infectivity potentiator (mip), gyrase subunit A (gyrA), ribosomal polymerase B (rpoB) and RNase P (rnpB) genes confirmed that the three isolates were distinct from recognized species of the genus Legionella. Phenotypic characterization of strain W03-356(T) based on fatty acid profiles confirmed that it was closely related to Legionella rubrilucens ATCC 35304(T) and Legionella pneumophila ATCC 33152(T), but distinct from other species of the genus Legionella. Serotyping of the isolates showed that they were distinct from all recognized species of the genus Legionella. Strains W03-356(T), W03-357 and W03-359 are thus considered to represent a novel species of the genus Legionella, for which the name Legionella dresdenensis sp. nov. is proposed. The type strain is W03-356(T) (=DSM 19488(T)=NCTC 13409(T)). PMID- 20008106 TI - Molecular systematics of marine gregarines (Apicomplexa) from North-eastern Pacific polychaetes and nemerteans, with descriptions of three novel species: Lecudina phyllochaetopteri sp. nov., Difficilina tubulani sp. nov. and Difficilina paranemertis sp. nov. AB - Most eugregarine apicomplexans infecting the intestines of marine invertebrates have been described within the family Lecudinidae and the type genus Lecudina. The diversity of these parasites is vast and poorly understood and only a tiny number of species has been characterized at the molecular phylogenetic level. DNA sequences coupled with high-resolution micrographs of trophozoites provide an efficient and precise approach for delimiting gregarine lineages from one another and also facilitate our overall understanding of gregarine biodiversity. In this study, phylogenetic analyses of small subunit (SSU) rDNA sequences from five (uncultivated) gregarines isolated from polychaetes and nemerteans in the North eastern Pacific Ocean are presented. Lecudina phyllochaetopteri sp. nov. was isolated from the intestines of the parchment tubeworm Phyllochaetopterus prolifica (Polychaeta). Lecudina longissima and Lecudina polymorpha were both isolated from the intestines of Lumbrineris japonica (Polychaeta). Difficilina tubulani sp. nov. was isolated from the nemertean Tubulanus polymorpha and Difficilina paranemertis sp. nov. was isolated from the nemertean Paranemertes peregrina. This is the first report of molecular sequence data from gregarines that infect nemerteans. The two novel species of the genus Difficilina described in this study formed a strongly supported clade in the phylogenetic analyses. This Difficilina clade formed the sister group to a robust subclade of lecudinids consisting of Lecudina longissima, Lecudina phyllochaetopteri sp. nov. (which lacked epicytic folds), Lecudina tuzetae, species of the genus Lankesteria and several sequences derived from previous environmental DNA surveys of marine biodiversity. PMID- 20008107 TI - Methanospirillum lacunae sp. nov., a methane-producing archaeon isolated from a puddly soil, and emended descriptions of the genus Methanospirillum and Methanospirillum hungatei. AB - A mesophilic, hydrogenotrophic methanogen, designated strain Ki8-1(T), was isolated from soil. Cells were strictly anaerobic, Gram-stain-negative, non sporulating, motile by means of a single flagellum or tufted flagella, and curved or wavy rod-shaped (11-25 MUm long). The temperature and pH for optimum growth were 30 degrees C and 7.5. The strain grew best in basal medium without the addition of NaCl. Methane was produced from H(2) and formate. Acetate or yeast extract was required for growth. The G+C content of the genomic DNA of strain Ki8 1(T) was 45.3 mol%. Phylogenetic analysis based on 16S rRNA gene sequences revealed that strain Ki8-1(T) was a member of the genus Methanospirillum and showed 95.1% sequence similarity to Methanospirillum hungatei NBRC 100397(T). On the basis of its phenotypic characteristics and phylogenetic position, strain Ki8 1(T) is considered to represent a novel species of the genus Methanospirillum, for which the name Methanospirillum lacunae sp. nov. is proposed. The type strain is Ki8-1(T) (NBRC 104920(T) =JCM 16384(T) =DSM 22751(T)). Emended descriptions of the genus Methanospirillum and of Methanospirillum hungatei are also provided. PMID- 20008108 TI - Bacteroides faecis sp. nov., isolated from human faeces. AB - Two anaerobic, Gram-negative, non-motile and non-spore-forming bacterial strains, designated MAJ27(T) and MAJ26, were isolated from human faeces. Both isolates grew optimally at 37 degrees C, were oxidase- and catalase-negative, were sensitive to bile and produced acid from fermentation of several substrates, including glucose. A study based on 16S rRNA gene sequences showed that both isolates were closely related to type strains of species of the genus Bacteroides. Comparisons of the isolates with Bacteroides thetaiotaomicron VPI 5482(T) and Bacteroides finegoldii JCM 13345(T) showed high levels of 16S rRNA gene sequence similarity (98.6-98.7 and 96.9-97.0 %, respectively), but low levels of DNA-DNA relatedness (<=22 %). The DNA G+C content (42.7+/-1 mol%) and the major fatty acid (anteiso-C(15 : 0), 39.3-42.5 %) supported the assignment of the isolates to the genus Bacteroides. Based on phenotypic, chemotaxonomic, genotypic and phylogenetic studies, we propose that strains MAJ27(T) and MAJ26 be classified as representing a novel species, Bacteroides faecis sp. nov. The type strain is MAJ27(T) (=KCTC 5823(T)=JCM 16478(T)). PMID- 20008109 TI - Dyadobacter soli sp. nov., a starch-degrading bacterium isolated from farm soil. AB - A Gram-negative, non-motile, aerobic bacterial strain, designated MJ20(T), was isolated from farm soil near Daejeon (South Korea) and was characterized taxonomically by using a polyphasic approach. Comparative 16S rRNA gene sequence analysis showed that strain MJ20(T) belongs to the family Cytophagaceae, class Sphingobacteria, and was related most closely to Dyadobacter fermentans DSM 18053(T) (98.9 % sequence similarity), Dyadobacter beijingensis JCM 14200(T) (98.0 %) and Dyadobacter ginsengisoli KCTC 12589(T) (96.4 %). The G+C content of the genomic DNA of strain MJ20(T) was 48.5 mol%. The detection of MK-7 as the predominant menaquinone and a fatty acid profile with summed feature 4 (C(16 : 1)omega7c and/or iso-C(15 : 0) 2-OH), iso-C(15 : 0), C(16 : 0) and C(16 : 1)omega5c as major components supported the affiliation of strain MJ20(T) to the genus Dyadobacter. The new isolate exhibited relatively low levels of DNA-DNA relatedness with respect to D. fermentans DSM 18053(T) (mean+/-SD of three determinations, 47+/-7 %) and D. beijingensis JCM 14200(T) (38+/-8 %). On the basis of its phenotypic and genotypic properties together with phylogenetic distinctiveness, strain MJ20(T) (=KCTC 22481(T) =JCM 16232(T)) should be classified in the genus Dyadobacter as the type strain of a novel species, for which the name Dyadobacter soli sp. nov. is proposed. PMID- 20008110 TI - Siphonobacter aquaeclarae gen. nov., sp. nov., a novel member of the family 'Flexibacteraceae', phylum Bacteroidetes. AB - A Gram-negative bacterium, designated P2(T), was isolated from the biofilm developed on the inner surface of an ultrapure cooling water system in a Hungarian power plant and was characterized by a polyphasic approach. Phylogenetic analysis based on 16S rRNA gene sequences revealed that strain P2(T) was affiliated with the family 'Flexibacteraceae' in the phylum Bacteroidetes. Its closest relative was Flectobacillus lacus CL-GP79(T) (88.7 % 16S rRNA gene sequence similarity) followed by Arcicella rosea TW5(T) (86.5 %), Arcicella aquatica NO-502(T) (86.4 %), Flectobacillus roseus GFA-11(T) (86.3 %) and Flectobacillus major DSM 103(T) (85.4 %). Cells of strain P2(T) were facultatively anaerobic, non-motile rods. The major fatty acids were C(16 : 1)omega5c (42.5 %), iso-C(15 : 0) 2-OH (17.2 %), iso-C(17 : 0) 3-OH (16.1 %) and iso-C(15 : 0) (8.5 %). The major menaquinone was MK-7 and the predominant polar lipid was phosphatidylethanolamine. The DNA G+C content was 54.5 mol%. Thus, the phenotypic and genotypic analyses clearly showed that strain P2(T) is considerably different from members of other genera in the family 'Flexibacteraceae'. Based on these results, it is concluded that strain P2(T) represents a novel species in a new genus, for which the name Siphonobacter aquaeclarae gen. nov., sp. nov. is proposed, with type strain P2(T) (=DSM 21668(T) =NCAIM B 02328(T)). PMID- 20008111 TI - Mesorhizobium robiniae sp. nov., isolated from root nodules of Robinia pseudoacacia. AB - Previously, five rhizobial strains isolated from root nodules of Robinia pseudoacacia were assigned to the same genospecies on the basis of identical 16S rRNA gene sequences and phylogenetic analyses of the nodA, nodC and nifH genes, in which the five isolates formed a well-supported group that excluded other sequences found in public databases. In this study, the 16S rRNA gene sequence similarities between the isolates and Mesorhizobium mediterraneum UPM-Ca36(T) and Mesorhizobium temperatum SDW018(T) were 99.5 and 99.6 %, respectively. The five isolates were also different from defined Mesorhizobium species using ERIC fingerprint profiles and they formed a novel Mesorhizobium lineage in phylogenetic analyses of recA and atpD gene sequences. DNA-DNA relatedness values between the representative strain, CCNWYC 115(T), and type strains of defined Mesorhizobium species were found to be lower than 47.5 %. These results indicated that the isolates represented a novel genomic species. Therefore, a novel species, Mesorhizobium robiniae sp. nov., is proposed, with type strain CCNWYC 115(T) (=ACCC 14543(T) =HAMBI 3082(T)). Strain CCNWYC 115(T) can form effective nodules only on its original host. PMID- 20008112 TI - Corynebacterium canis sp. nov., isolated from a wound infection caused by a dog bite. AB - A non-lipophilic, coryneform bacterium isolated from a patient's wound caused by a dog bite was characterized by phenotypic, chemotaxonomic and molecular genetic methods. Chemotaxonomic features suggested assignment of the unknown bacterium to the genus Corynebacterium. The isolate exhibited the following unusual features, which made it possible to phenotypically differentiate it from all other medically relevant corynebacteria: the Gram stain showed some very filamentous rods (>15 MUm in length); some cells exhibited branching; colonies were domed and adherent to agar; the micro-organism was positive for pyrazinamidase, beta glucosidase, alpha-glucosidase and trypsin but negative for beta-galactosidase. 16S rRNA gene sequencing and partial rpoB gene sequencing showed that the closest phylogenetic relative, Corynebacterium freiburgense, exhibited more than 1.9 % and 17.9 % divergence with the unknown bacterium, respectively. Based on both phenotypic and molecular genetic data, it is proposed that the isolate should be classified as a novel species, Corynebacterium canis sp. nov., with the type strain 1170(T) (=CCUG 58627(T) =DSM 45402(T)). PMID- 20008113 TI - Williamsia faeni sp. nov., an actinomycete isolated from a hay meadow. AB - The taxonomic status of an actinomycete isolated from soil collected from a hay meadow was determined using a polyphasic approach. The strain, designated N1350(T), had morphological and chemotaxonomic properties consistent with its classification in the genus Williamsia and formed a distinct phyletic line within the clade comprising the type strains of species of the genus Williamsia in the 16S rRNA gene tree. Strain N1350(T) shared highest 16S rRNA gene sequence similarities with Williamsia marianensis MT8(T) (98.1 %) and Williamsia muralis MA140-96(T) (98.3 %). However, strain N1350(T) was readily distinguished from the type strains of Williamsia species using a combination of phenotypic properties. On the basis of these data, strain N1350(T) is considered to represent a novel species of the genus Williamsia. The name proposed for this taxon is Williamsia faeni sp. nov., with the type strain N1350(T) (=DSM 45372(T) =NCIMB 14575(T) =NRRL B-24794(T)). PMID- 20008114 TI - Increase in passive stiffness at reduced airway smooth muscle length: potential impact on airway responsiveness. AB - The amplitude of strain in airway smooth muscle (ASM) produced by oscillatory perturbations such as tidal breathing or deep inspiration (DI) influences the force loss in the muscle and is therefore a key determinant of the bronchoprotective and bronchodilatory effects of these breathing maneuvers. The stiffness of unstimulated ASM (passive stiffness) directly influences the amplitude of strain. The nature of the passive stiffness is, however, not clear. In this study, we measured the passive stiffness of ovine ASM at different muscle lengths (relative to in situ length, which was used as a reference length, L(ref)) and states of adaptation to gain insights into the origin of this muscle property. The results showed that the passive stiffness was relatively independent of muscle length, possessing a constant plateau value over a length range from 0.62 to 1.25 L(ref). Following a halving of ASM length, passive stiffness decreased substantially (by 71%) but redeveloped over time ( approximately 30 min) at the shorter length to reach 65% of the stiffness value at L(ref), provided that the muscle was stimulated to contract at least once over a approximately 30-min period. The redevelopment and maintenance of passive stiffness were dependent on the presence of Ca(2+) but unaffected by latrunculin B, an inhibitor of actin filament polymerization. The maintenance of passive stiffness was also not affected by blocking myosin cross-bridge cycling using a myosin light chain kinase inhibitor or by blocking the Rho-Rho kinase (RhoK) pathway using a RhoK inhibitor. Our results suggest that the passive stiffness of ASM is labile and capable of redevelopment following length reduction. Redevelopment and maintenance of passive stiffness following muscle shortening could contribute to airway hyperresponsiveness by attenuating the airway wall strain induced by tidal breathing and DI. PMID- 20008115 TI - Modulation of epithelial sodium channel activity by lipopolysaccharide in alveolar type II cells: involvement of purinergic signaling. AB - Pseudomonas aeruginosa is a gram-negative bacterium that causes chronic infection in cystic fibrosis patients. We reported recently that P. aeruginosa modulates epithelial Na(+) channel (ENaC) expression in experimental chronic pneumonia models. For this reason, we tested whether LPS from P. aeruginosa alters ENaC expression and activity in alveolar epithelial cells. We found that LPS induces a approximately 60% decrease of ENaC apical current without significant changes in intracellular ENaC or surface protein expression. Because a growing body of evidence reports a key role for extracellular nucleotides in regulation of ion channels, we evaluated the possibility that modulation of ENaC activity by LPS involves extracellular ATP signaling. We found that alveolar epithelial cells release ATP upon LPS stimulation and that pretreatment with suramin, a P2Y(2) purinergic receptor antagonist, inhibited the effect of LPS on ENaC. Furthermore, ET-18-OCH3, a PLC inhibitor, and Go-6976, a PKC inhibitor, were able to partially prevent ENaC inhibition by LPS, suggesting that the actions of LPS on ENaC current were mediated, in part, by the PKC and PLC pathways. Together, these findings demonstrate an important role of extracellular ATP signaling in the response of epithelial cells to LPS. PMID- 20008116 TI - Bone marrow-derived angiogenic cells restore lung alveolar and vascular structure after neonatal hyperoxia in infant mice. AB - Neonatal hyperoxia impairs vascular and alveolar growth in mice and decreases endothelial progenitor cells. To determine the role of bone marrow-derived cells in restoration of neonatal lung structure after injury, we studied a novel bone marrow myeloid progenitor cell population from Tie2-green fluorescent protein (GFP) transgenic mice (bone marrow-derived angiogenic cells; BMDAC). We hypothesized that treatment with BMDAC would restore normal lung structure in infant mice during recovery from neonatal hyperoxia. Neonatal mice (1-day-old) were exposed to 80% oxygen for 10 days. BMDACs (1 x 10(5)), embryonic endothelial progenitor cells, mouse embryonic fibroblasts (control), or saline were then injected into the pulmonary circulation. At 21 days of age, saline-treated mice had enlarged alveoli, reduced septation, and a reduction in vascular density. In contrast, mice treated with BMDAC had complete restoration of lung structure that was indistinguishable from room air controls. BMDAC comprised 12% of distal lung cells localized to pulmonary vessels or alveolar type II (AT2) cells and persist (8.8%) for 8 wk postinjection. Coculture of AT2 cells or lung endothelial cells (luEC) with BMDAC augmented AT2 and luEC cell growth in vitro. We conclude that treatment with BMDAC after neonatal hyperoxia restores lung structure in this model of bronchopulmonary dysplasia. PMID- 20008118 TI - alpha1,2-Fucosylated and beta-N-acetylgalactosaminylated prostate-specific antigen as an efficient marker of prostatic cancer. AB - A prostate-specific antigen (PSA) is widely used as a diagnostic marker for prostate cancer (PC) because of its high specificity. However, elevated serum PSA does not occur only in PC but also in benign prostatic hyperplasia (BPH). Since the structural changes of N-glycans during carcinogenesis are common phenomena, we investigated whether PC-specific N-glycans are linked to PSA. We first analyzed the carbohydrate structures of PSA derived from seminal fluid, serum of BPH and PC patients, and PC cell line, namely, LNCaP using eight lectin immobilized columns and then with enzyme-linked immunosorbent assay (ELISA). The fraction of serum PSA from PC patients bound to both Fucalpha1-2Gal and betaGalNAc binding Trichosanthes japonica agglutinin-II (TJA-II) column, while that from BPH patients did not exhibit this binding ability, thereby implying that there is elevated expression of alpha1,2-fucosylation and beta-N acetylgalactosaminylation of PSA during carcinogenesis. We then performed a real time polymerase chain reaction (PCR) and confirmed that these structural changes were responsible for the elevated expression of fucosyltransferase I (FUT1) and beta-N-acetylgalactosaminyltransferase 4(B4GALNT4). Second, we measured TJA-II bound PSA contents and the binding ratios of TJA-II column chromatography in serum PSA samples from 40 patients of both PC and BPH. The results indicated that both TJA-II-bound PSA content and TJA-II binding ratios (%) could be used to discriminate between PC and BPH with more than 95% probability, and TJA-II-bound PSA can be regarded as a potential marker of PC. PMID- 20008117 TI - Modulation of endocytic trafficking and apical stability of CFTR in primary human airway epithelial cultures. AB - CFTR is a highly regulated apical chloride channel of epithelial cells that is mutated in cystic fibrosis (CF). In this study, we characterized the apical stability and intracellular trafficking of wild-type and mutant CFTR in its native environment, i.e., highly differentiated primary human airway epithelial (HAE) cultures. We labeled the apical pool of CFTR and subsequently visualized the protein in intracellular compartments. CFTR moved from the apical surface to endosomes and then efficiently recycled back to the surface. CFTR endocytosis occurred more slowly in polarized than in nonpolarized HAE cells or in a polarized epithelial cell line. The most common mutation in CF, DeltaF508 CFTR, was rescued from endoplasmic reticulum retention by low-temperature incubation but transited from the apical membrane to endocytic compartments more rapidly and recycled less efficiently than wild-type CFTR. Incubation with small-molecule correctors resulted in DeltaF508 CFTR at the apical membrane but did not restore apical stability. To stabilize the mutant protein at the apical membrane, we found that the dynamin inhibitor Dynasore and the cholesterol-extracting agent cyclodextrin dramatically reduced internalization of DeltaF508, whereas the proteasomal inhibitor MG-132 completely blocked endocytosis of DeltaF508. On examination of intrinsic properties of CFTR that may affect its apical stability, we found that N-linked oligosaccharides were not necessary for transport to the apical membrane but were required for efficient apical recycling and, therefore, influenced the turnover of surface CFTR. Thus apical stability of CFTR in its native environment is affected by properties of the protein and modulation of endocytic trafficking. PMID- 20008119 TI - Use of dansyl-cholestanol as a probe of cholesterol behavior in membranes of living cells. AB - While plasma membrane cholesterol-rich microdomains play a role in cholesterol trafficking, little is known about the appearance and dynamics of cholesterol through these domains in living cells. The fluorescent cholesterol analog 6 dansyl-cholestanol (DChol), its biochemical fractionation, and confocal imaging of L-cell fibroblasts contributed the following new insights: i) fluorescence properties of DChol were sensitive to microenvironment polarity and mobility; (ii) DChol taken up by L-cell fibroblasts was distributed similarly as cholesterol and preferentially into cholesterol-rich vs. -poor microdomains resolved by affinity chromatography of purified plasma membranes; iii) DChol reported similar polarity (dielectric constant near 18) but higher mobility near phospholipid polar head group region for cholesterol in purified cholesterol-rich versus -poor microdomains; and iv) real-time confocal imaging, quantitative colocalization analysis, and fluorescence resonance energy transfer with cholesterol-rich and -poor microdomain markers confirmed that DChol preferentially localized in plasma membrane cholesterol-rich microdomains of living cells. Thus, DChol sensed a unique, relatively more mobile microenvironment for cholesterol in plasma membrane cholesterol-rich microdomains, consistent with the known, more rapid exchange dynamics of cholesterol from cholesterol-rich than -poor microdomains. PMID- 20008120 TI - Lipids and lipid metabolism in the eye. PMID- 20008122 TI - Gradient, contact-free volume transfers minimize compound loss in dose-response experiments. AB - More accurate dose-response curves can be constructed by eliminating aqueous serial dilution of compounds. Traditional serial dilutions that use aqueous diluents can result in errors in dose-response values of up to 4 orders of magnitude for a significant percentage of a compound library. When DMSO is used as the diluent, the errors are reduced but not eliminated. The authors use acoustic drop ejection (ADE) to transfer different volumes of model library compounds, directly creating a concentration gradient series in the receiver assay plate. Sample losses and contamination associated with compound handling are therefore avoided or minimized, particularly in the case of less water soluble compounds. ADE is particularly well suited for assay miniaturization, but gradient volume dispensing is not limited to miniaturized applications. PMID- 20008123 TI - Maximizing RNA yield from archival renal tumors and optimizing gene expression analysis. AB - Formalin-fixed, paraffin-embedded tissues are widely available for gene expression analysis using TaqMan PCR. Five methods, including 4 commercial kits, for recovering RNA from paraffin-embedded renal tumor tissue were compared. The MasterPure kit from Epicentre produced the highest RNA yield. However, the difference in RNA yield between the kit from Epicenter and Invitrogen's TRIzol method was not significant. Using the top 3 RNA isolation methods, the manufacturers' protocols were modified to include an overnight Proteinase K digestion. Overnight protein digestion resulted in a significant increase in RNA yield. To optimize the reverse transcription reaction, conventional reverse transcription with random oligonucleotide primers was compared to reverse transcription using primers specific for genes of interest. Reverse transcription using gene-specific primers significantly increased the quantity of cDNA detectable by TaqMan PCR. Therefore, expression profiling of formalin-fixed, paraffin-embedded tissue using TaqMan qPCR can be optimized by using the MasterPure RNA isolation kit modified to include an overnight Proteinase K digestion and gene-specific primers during the reverse transcription. PMID- 20008121 TI - Bile acids: analysis in biological fluids and tissues. AB - The formation of bile acids/bile alcohols is of major importance for the maintenance of cholesterol homeostasis. Besides their functions in lipid absorption, bile acids/bile alcohols are regulatory molecules for a number of metabolic processes. Their effects are structure-dependent, and numerous metabolic conversions result in a complex mixture of biologically active and inactive forms. Advanced methods are required to characterize and quantify individual bile acids in these mixtures. A combination of such analyses with analyses of the proteome will be required for a better understanding of mechanisms of action and nature of endogenous ligands. Mass spectrometry is the basic detection technique for effluents from chromatographic columns. Capillary liquid chromatography-mass spectrometry with electrospray ionization provides the highest sensitivity in metabolome analysis. Classical gas chromatography-mass spectrometry is less sensitive but offers extensive structure-dependent fragmentation increasing the specificity in analyses of isobaric isomers of unconjugated bile acids. Depending on the nature of the bile acid/bile alcohol mixture and the range of concentration of individuals, different sample preparation sequences, from simple extractions to group separations and derivatizations, are applicable. We review the methods currently available for the analysis of bile acids in biological fluids and tissues, with emphasis on the combination of liquid and gas phase chromatography with mass spectrometry. PMID- 20008124 TI - High-content screening: a decade of evolution. AB - In the past decade, high-content screening has become a highly developed approach to obtaining richly descriptive quantitative phenotypic data using automated microscopy. From early use in drug screening, the technique has evolved to embrace a diverse range of applications in both academic and industrial sectors and is now widely recognized as providing an efficient and effective approach to large-scale programs investigating cell biology in situ and in context. PMID- 20008125 TI - Screening for inhibitors of low-affinity epigenetic peptide-protein interactions: an AlphaScreen-based assay for antagonists of methyl-lysine binding proteins. AB - The histone code comprises many posttranslational modifications that occur mainly in histone tail peptides. The identity and location of these marks are read by a variety of histone-binding proteins that are emerging as important regulators of cellular differentiation and development and are increasingly being implicated in numerous disease states. The authors describe the development of the first high throughput screening assay for the discovery of inhibitors of methyl-lysine binding proteins that will be used to initiate a full-scale discovery effort for this broad target class. They focus on the development of an AlphaScreen-based assay for malignant brain tumor (MBT) domain-containing proteins, which bind to the lower methylation states of lysine residues present in histone tail peptides. This assay takes advantage of the avidity of the AlphaScreen beads to clear the hurdle to assay development presented by the low micromolar binding constants of the histone binding proteins for their cognate peptides. The assay is applicable to other families of methyl-lysine binding proteins, and it has the potential to be used in screening efforts toward the discovery of novel small molecules with utility as research tools for cellular reprogramming and ultimately drug discovery. PMID- 20008126 TI - Identification of a small GTPase inhibitor using a high-throughput flow cytometry bead-based multiplex assay. AB - Small GTPases are key regulators of cellular activity and represent novel targets for the treatment of human diseases using small-molecule inhibitors. The authors describe a multiplex, flow cytometry bead-based assay for the identification and characterization of inhibitors or activators of small GTPases. Six different glutathione-S-transferase (GST)-tagged small GTPases were bound to glutathione beads, each labeled with a different red fluorescence intensity. Subsequently, beads bearing different GTPase were mixed and dispensed into 384-well plates with test compounds, and fluorescent-guanosine triphosphate (GTP) binding was used as the readout. This novel multiplex assay allowed the authors to screen a library of almost 200,000 compounds and identify more than 1200 positive compounds, which were further verified by dose-response analyses, using 6- to 8-plex assays. After the elimination of false-positive and false-negative compounds, several small molecule families with opposing effects on GTP binding activity were identified. The authors detail the characterization of MLS000532223, a general inhibitor that prevents GTP binding to several GTPases in a dose-dependent manner and is active in biochemical and cell-based secondary assays. Live-cell imaging and confocal microscopy studies revealed the inhibitor-induced actin reorganization and cell morphology changes, characteristic of Rho GTPases inhibition. Thus, high throughput screening via flow cytometry provides a strategy for identifying novel compounds that are active against small GTPases. PMID- 20008128 TI - The origin of renal fibroblasts and progression of kidney disease. PMID- 20008127 TI - Fate tracing reveals the pericyte and not epithelial origin of myofibroblasts in kidney fibrosis. AB - Understanding the origin of myofibroblasts in kidney is of great interest because these cells are responsible for scar formation in fibrotic kidney disease. Recent studies suggest epithelial cells are an important source of myofibroblasts through a process described as the epithelial-to-mesenchymal transition; however, confirmatory studies in vivo are lacking. To quantitatively assess the contribution of renal epithelial cells to myofibroblasts, we used Cre/Lox techniques to genetically label and fate map renal epithelia in models of kidney fibrosis. Genetically labeled primary proximal epithelial cells cultured in vitro from these mice readily induce markers of myofibroblasts after transforming growth factor beta(1) treatment. However, using either red fluorescent protein or beta-galactosidase as fate markers, we found no evidence that epithelial cells migrate outside of the tubular basement membrane and differentiate into interstitial myofibroblasts in vivo. Thus, although renal epithelial cells can acquire mesenchymal markers in vitro, they do not directly contribute to interstitial myofibroblast cells in vivo. Lineage analysis shows that during nephrogenesis, FoxD1-positive((+)) mesenchymal cells give rise to adult CD73(+), platelet derived growth factor receptor beta(+), smooth muscle actin-negative interstitial pericytes, and these FoxD1-derivative interstitial cells expand and differentiate into smooth muscle actin(+) myofibroblasts during fibrosis, accounting for a large majority of myofibroblasts. These data indicate that therapeutic strategies directly targeting pericyte differentiation in vivo may productively impact fibrotic kidney disease. PMID- 20008129 TI - Antigen-specific induction of osteopontin contributes to the chronification of allergic contact dermatitis. AB - Allergic contact dermatitis is a T cell-mediated immune response, which in its relapsing chronic form is of high socioeconomic impact. The phosphoglycoprotein osteopontin (OPN) has chemotactic and Th1 cytokine functions and in various models is essential for robust T cell-mediated immunity. Here we demonstrate that OPN is abundantly expressed by both effector T cells and keratinocytes in allergic contact dermatitis lesions. T cells from nickel-allergic donors secrete high levels of OPN following antigen-specific stimulation. OPN may substitute for missing IFN-gamma secretion in T effector cells because low IFN-gamma-producing T cell clones secrete high levels of OPN, and OPN down-modulates their interleukin 4 expression. Furthermore, interferon-gamma from T effector cells augments OPN in allergic contact dermatitis by inducing OPN in keratinocytes, which in turn polarizes dendritic cells and attracts inflammatory cells. In the murine contact hypersensitivity (CHS) model for allergic contact dermatitis, OPN is strongly induced in antigen-specific proliferating T cells, and OPN null mice display a reduced chronic CHS inflammatory response due to a decreased influx of effector T cells. Importantly, because of its function for chronic allergic contact dermatitis, OPN may well be a therapeutic target, because anti-OPN antibody treatment in part suppresses established chronic CHS. PMID- 20008130 TI - Downstream targets of homeobox gene HLX show altered expression in human idiopathic fetal growth restriction. AB - Fetal growth restriction (FGR), a clinically significant pregnancy disorder, is poorly understood at the molecular level. This study investigates idiopathic FGR associated with placental insufficiency. Previously, we showed that the homeobox gene HLX is expressed in placental trophoblast cells and that HLX expression is significantly decreased in human idiopathic FGR. Here, we used the novel approach of identifying downstream targets of HLX in cell culture to detect potentially important genes involved in idiopathic FGR. Downstream targets were revealed by decreasing HLX expression in cultured trophoblast cells with HLX-specific small interfering RNAs to model human idiopathic FGR and comparing these levels with controls using a real-time PCR-based gene profiling system. Changes in candidate HLX target mRNA levels were verified in an independent trophoblast cell line, and candidate target gene expression was assessed in human idiopathic FGR-affected placentae (n = 25) compared with gestation-matched controls (n = 25). The downstream targets RB1 and MYC, cell cycle regulatory genes, showed significantly increased mRNA levels in FGR-affected tissues compared with gestation-matched controls, whereas CCNB1, ELK1, JUN, and CDKN1 showed significantly decreased mRNA levels (n = 25, P < 0.001, t-test). The changes for RB1 and CDKN1C were verified by Western blot analysis in FGR-affected placentae compared with gestation matched controls (n = 6). We conclude that cell cycle regulatory genes RB1, MYC, CCNB1, ELK1, JUN, and CDKN1C, which control important trophoblast cell functions, are targets of HLX. PMID- 20008131 TI - Platelets control leukocyte recruitment in a murine model of cutaneous arthus reaction. AB - Platelets have been shown to be important in inflammation, but their role in the cutaneous Arthus reaction remains unclear. To assess the role of platelets in this pathogenetic process, the cutaneous Arthus reaction was examined in wild type mice and mice lacking E-selectin, P-selectin, or P-selectin glycoprotein ligand-1 (PSGL-1) with or without platelet depletion by busulfan, a bone marrow precursor cell-specific toxin. Edema and hemorrhage induced by immune complex challenge significantly decreased in busulfan-treated wild-type mice compared with untreated mice. Busulfan treatment did not affect edema and hemorrhage in P selectin- or PSGL-1-deficient mice, suggesting that the effect by busulfan is dependent on P-selectin and PSGL-1 expression. The inhibited edema and hemorrhage paralleled reduced infiltration of neutrophils and mast cells and reduced levels of circulating platelets. Increased cutaneous production of interleukin-6, tumor necrosis factor-alpha, and platelet-derived chemokines during Arthus reaction was inhibited in busulfan-treated wild-type mice relative to untreated mice, which paralleled the reduction in cutaneous inflammation. Flow cytometric analysis showed that immune complex challenge generated blood platelet-leukocyte aggregates that decreased by busulfan treatment. In thrombocytopenic mice, the cutaneous inflammation after immune complex challenge was restored by platelet infusion. These results suggest that platelets induce leukocyte recruitment into skin by forming platelet-leukocyte aggregates and secreting chemokines at inflamed sites, mainly through the interaction of P-selectin on platelets with PSGL-1 on leukocytes. PMID- 20008132 TI - Initiation of acquired immunity in the lungs of mice lacking lymph nodes after infection with aerosolized Mycobacterium tuberculosis. AB - Recent evidence points to lung draining lymph nodes as the site that initiates the immune response in mice infected with aerosolized Mycobacterium tuberculosis. Here we expanded these studies and showed that infection of mice that lack lymph nodes with aerosolized M. tuberculosis results in a massive mononuclear cell infiltrate in the lungs within 14 days postinfection. This infiltration clearly resembles an expansion of the bronchus-associated lymphoid tissue. As expected, no bronchus-associated lymphoid tissue was observed in M. tuberculosis-infected wild-type control mice. Importantly, acquired specific immune response to M. tuberculosis antigens could be detected in lung lymphocytes harvested from mice lacking lymph nodes as early as 14 days postinfection. In addition, the bacterial burden in these mice was indistinguishable from that observed in wild-type C57BL/6 control mice. These results indicate that in the absence of lymph nodes, priming of the immune response occurs in the lung tissues after infection of mice with aerosolized M. tuberculosis and clearly illustrate the enormous plasticity of the immune system to develop resistance to foreign pathogens. PMID- 20008133 TI - Endometrial cancer side-population cells show prominent migration and have a potential to differentiate into the mesenchymal cell lineage. AB - Cancer stem-like cell subpopulations, referred to as "side-population" (SP) cells, have been identified in several tumors based on their ability to efflux the fluorescent dye Hoechst 33342. Although SP cells have been identified in the normal human endometrium and endometrial cancer, little is known about their characteristics. In this study, we isolated and characterized the SP cells in human endometrial cancer cells and in rat endometrial cells expressing oncogenic human K-Ras protein. These SP cells showed i) reduction in the expression levels of differentiation markers; ii) long-term proliferative capacity of the cell cultures; iii) self-renewal capacity in vitro; iv) enhancement of migration, lamellipodia, and uropodia formation; and v) enhanced tumorigenicity. In nude mice, SP cells formed large, invasive tumors, which were composed of both tumor cells and stromal-like cells with enriched extracellular matrix. The expression levels of vimentin, alpha-smooth muscle actin, and collagen III were enhanced in SP tumors compared with the levels in non-SP tumors. In addition, analysis of microdissected samples and fluorescence in situ hybridization of Hec1-SP-tumors showed that the stromal-like cells with enriched extracellular matrix contained human DNA, confirming that the stromal-like cells were derived from the inoculated cells. Moreober, in a Matrigel assay, SP cells differentiated into alpha-smooth muscle actin-expressing cells. These findings demonstrate that SP cells have cancer stem-like cell features, including the potential to differentiate into the mesenchymal cell lineage. PMID- 20008134 TI - Tissue factor-deficiency and protease activated receptor-1-deficiency reduce inflammation elicited by diet-induced steatohepatitis in mice. AB - Altered hepatic lipid homeostasis, hepatocellular injury, and inflammation are features of nonalcoholic steatohepatitis, which contributes significantly to liver-related morbidity and mortality in the Western population. A collection of inflammatory mediators have been implicated in the pathogenesis of steatohepatitis in mouse models. However, the pathways essential for coordination and amplification of hepatic inflammation and injury caused by steatosis are not completely understood. We tested the hypothesis that tissue factor (TF)-dependent thrombin generation and the thrombin receptor protease activated receptor-1 (PAR 1) contribute to liver inflammation induced by steatosis in mice. Wild-type C57Bl/6J mice fed a diet deficient in methionine and choline for 2 weeks manifested steatohepatitis characterized by increased serum alanine aminotransferase activity, macrovesicular hepatic steatosis, hepatic inflammatory gene expression, and lobular inflammation. Steatohepatitis progression was associated with thrombin generation and hepatic fibrin deposition. Coagulation cascade activation was significantly reduced in low TF mice, which express 1% of normal TF levels. Hepatic triglyceride accumulation was not affected in low TF mice or PAR-1-deficient mice. In contrast, biomarkers of hepatocellular injury, inflammatory gene induction, and hepatic accumulation of macrophages and neutrophils were greatly reduced by TF-deficiency and PAR-1-deficiency. The results suggest that TF-dependent thrombin generation and activation of PAR-1 amplify hepatic inflammation and injury during the pathogenesis of steatohepatitis. PMID- 20008135 TI - Cellular plasticity of inflammatory myeloid cells in the peritoneal foreign body response. AB - Implantation of sterile foreign objects in the peritoneal cavity of an animal initiates an inflammatory response and results in encapsulation of the objects by bone marrow-derived cells. Over time, a multilayered tissue capsule develops with abundant myofibroblasts embedded in extracellular matrix. The present study used the transgenic MacGreen mouse to characterize the time-dependent accumulation of monocyte subsets and neutrophilic granulocytes in the inflammatory infiltrate and within the tissue capsule by their differential expression of the csf1r-EGFP transgene, F4/80, and Ly6C. As the tissue capsule developed, enhanced green fluorescent protein-positive cells changed from rounded to spindle-shaped morphology and began to co-express the myofibroblast marker alpha-smooth muscle actin. Expression increased with time: at day 14, 11.13 +/- 0.67% of tissue capsule cells co-expressed these markers, compared with 50.77 +/- 12.85% of cells at day 28. The importance of monocyte/macrophages in tissue capsule development was confirmed by clodronate-encapsulated liposome removal, which resulted in almost complete abrogation of capsule development. These results confirm the importance of monocyte/macrophages in the tissue response to sterile foreign objects implanted in the peritoneal cavity. In addition, the in vivo plasticity of peritoneal macrophages and their ability to transdifferentiate from a myeloid to mesenchymal phenotype is demonstrated. PMID- 20008136 TI - WNT5A expression in ameloblastoma and its roles in regulating enamel epithelium tumorigenic behaviors. AB - Odontogenic tumors originate from the remains of migrating enamel epithelium after the completion of normal tooth genesis. These enamel epithelium remnants exhibit the ability to recapitulate the events that occur during tooth formation. Several lines of evidence suggest that aberrance in the signaling pathways similar to the ones that are used during tooth development, including the WNT pathway, might be the cause of odontogenic tumorigenesis and maintenance. In this study we demonstrated that WNT5A expression was intense in both the epithelial component of ameloblastomas, the most common epithelial odontogenic tumor, and in this tumor's likely precursor cell, the enamel epithelium located at the cervical loop of normal developing human tooth buds. Additionally, when WNT5A was overexpressed in enamel epithelium cells (LS-8), the clones expressing high levels of WNT5A (S) exhibited characteristics of tumorigenic cells, including growth factor independence, loss of anchorage dependence, loss of contact inhibition, and tumor formation in immunocompromised mice. Moreover, overexpression of WNT5A drastically increased LS-8 cell migration and actin reorganization when compared with controls. Suppression of endogenous WNT5A in LS 8 cells (AS) greatly impaired their migration and AS cells failed to form significant actin reorganization and membrane protrusion was rarely seen. Taken together, our data indicate that WNT5A signaling is important in modulating tumorigenic behaviors of enamel epithelium cells in ameloblastomas. PMID- 20008137 TI - The absence of interleukin-6 enhanced arsenite-induced renal injury by promoting autophagy of tubular epithelial cells with aberrant extracellular signal regulated kinase activation. AB - Sodium arsenite (NaAs)-induced autophagic cell death (ACD) of a mouse renal tubular epithelial cell line (mProx24), which expresses enhanced levels of interleukin-6 (IL-6), was reduced by the suppression of autophagy by 3 methyladenine or Atg7 knockdown. The inhibition of the IL-6/signal transducer and activator of transcription 3 (STAT3) signal pathway by anti-IL-6 antibody or a Jak2 inhibitor (AG490) exaggerated ACD of mProx24 cells after NaAs challenge, attenuating STAT3 activation and reciprocally enhancing extracellular signal regulated kinase (ERK) phosphorylation. In contrast, an ERK inhibitor, PD98059, reduced NaAs-induced ACD in mProx24 cells. Subcutaneous injection of NaAs (12.5 mg/kg) into BALB/c (wild-type) mice enhanced intrarenal expression of IL-6, mainly produced by tubular cells, and caused severe renal injury characterized by hemorrhages, acute tubular necrosis, cast formation, and brush border disappearance, with increases in serum urea nitrogen (blood urea nitrogen) and creatinine levels. In addition, IL-6-deficient (IL-6(-/-)) mice exhibited exaggerated histopathological changes with higher blood urea nitrogen and creatinine levels. Moreover, in IL-6(-/-) mice treated with NaAs, ACD in renal tubular cells was significantly augmented, along with diminished STAT3 activation and reciprocal enhancement of ERK signaling, compared with wild-type mice. Finally, the administration of exogenous IL-6 into wild-type mice significantly reduced NaAs-induced ACD along with diminished ERK activation and eventually alleviated acute renal dysfunction. Thus, IL-6/STAT3 signal pathway could inhibit ERK activation, a crucial step for ACD, eventually attenuating NaAs-induced renal dysfunction. PMID- 20008138 TI - Epithelial NF-kappaB enhances transmucosal fluid movement by altering tight junction protein composition after T cell activation. AB - In inflammatory bowel disease (IBD), aberrant activation of innate and adaptive immune responses enhances mucosal permeability through mechanisms not completely understood. To examine the role of epithelial nuclear factor (NF-kappaB) in IBD induced enhanced permeability, epithelial-specific IkappaBalpha mutant (NF-kappaB super repressor) transgenic (TG) mice were generated. NF-kB activation was inhibited in TG mice, relative to wild-type mice, following T cell-mediated immune cell activation using an anti-CD3 monoclonal antibody. Furthermore, epithelial NF-kappaB super repressor protein inhibited diarrhea and blocked changes in transepithelial resistance and transmucosal flux of alexa350 (0.35 kDa) and dextran3000 (3 kDa). In vivo perfusion loop studies in TG mice revealed reversed net water secretion and reduced lumenal flux of different molecular probes (bovine serum albumin, alexa350, and dextran3000). Cell-imaging and immunoblotting of low-density, detergent-insoluble membrane fractions confirmed that tight junction proteins (occludin, claudin-1 and zona occludens-1) are internalized through an NF-kappaB-dependent pathway. Taken together, these data suggest that IBD-associated diarrhea results from NF-kappaB-mediated tight junction protein internalization and increased paracellular permeability. Thus, reduction of epithelial NF-kappaB activation in IBD may repair defects in epithelial barrier function, reduce diarrhea, and limit protein (eg, serum albumin) losses. Epithelial NF-kappaB activation induced by mucosal T cells, therefore, actively plays a role in opening paracellular spaces to promote transmucosal fluid effux into the intestinal lumen. PMID- 20008139 TI - Nuclear beta-catenin induces an early liver progenitor phenotype in hepatocellular carcinoma and promotes tumor recurrence. AB - Transforming growth factor-beta cooperates with oncogenic Ras to activate nuclear beta-catenin during the epithelial to mesenchymal transition of hepatocytes, a process relevant in the progression of hepatocellular carcinoma (HCC). In this study we investigated the role of beta-catenin in the differentiation of murine, oncogene-targeted hepatocytes and in 133 human HCC patients scheduled for orthotopic liver transplantation. Transforming growth factor-beta caused dissociation of plasma membrane E-cadherin/beta-catenin complexes and accumulation of nuclear beta-catenin in Ras-transformed, but otherwise normal hepatocytes in p19(ARF)-/- mice. Both processes were inhibited by Smad7-mediated disruption of transforming growth factor-beta signaling. Overexpression of constitutively active beta-catenin resulted in high levels of CK19 and M2-PK, whereas ablation of beta-catenin by axin overexpression caused strong expression of CK8 and CK18. Therefore, nuclear beta-catenin resulted in dedifferentiation of neoplastic hepatocytes to immature progenitor cells, whereas loss of nuclear beta catenin led to a differentiated HCC phenotype. Poorly differentiated human HCC showed cytoplasmic redistribution or even loss of E-cadherin, suggesting epithelial to mesenchymal transition. Analysis of 133 HCC patient samples revealed that 58.6% of human HCC exhibited strong nuclear beta-catenin accumulation, which correlated with clinical features such as vascular invasion and recurrence of disease after orthotopic liver transplantation. These data suggest that activation of beta-catenin signaling causes dedifferentiation to malignant, immature hepatocyte progenitors and facilitates recurrence of human HCC after orthotopic liver transplantation. PMID- 20008140 TI - MYC high level gene amplification is a distinctive feature of angiosarcomas after irradiation or chronic lymphedema. AB - Angiosarcomas (AS) are rare vascular malignancies that arise either de novo as primary tumors or secondary to irradiation or chronic lymphedema. The cytogenetics of angiosarcomas are poorly characterized. We applied array comparative genomic hybridization as a screening method to identify recurrent alterations in 22 cases. Recurrent genetic alterations were identified only in secondary but not in primary AS. The most frequent recurrent alterations were high level amplifications on chromosome 8q24.21 (50%), followed by 10p12.33 (33%) and 5q35.3 (11%). Fluorescence in situ hybridization analysis in 28 primary and 33 secondary angiosarcomas (31 tumors secondary to irradiation, 2 tumors secondary to chronic lymphedema) confirmed high level amplification of MYC on chromosome 8q24.21 as a recurrent genetic alteration found exclusively in 55% of AS secondary to irradiation or chronic lymphedema, but not in primary AS. Amplification of MYC did not predispose to high grade morphology or increased cell turnover. In conclusion, despite their identical morphology, secondary AS are genetically different from primary AS and are characterized by a high frequency of high level amplifications of MYC. This finding may have implications both for the diagnosis and treatment of these tumors. PMID- 20008141 TI - Age-related vascular pathology in transgenic mice expressing presenilin 1 associated familial Alzheimer's disease mutations. AB - Mutations in the presenilin 1 (PS1) gene are the most commonly recognized cause of familial Alzheimer's disease (FAD). Besides senile plaques, neurofibrillary tangles, and neuronal loss, Alzheimer's disease (AD) is also accompanied by vascular pathology. Here we describe an age-related vascular pathology in two lines of PS1 FAD-mutant transgenic mice that mimics many features of the vascular pathology seen in AD. The pathology was especially prominent in the microvasculature whose vessels became thinned and irregular with the appearance of many abnormally looped vessels as well as string vessels. Stereologic assessments revealed a reduction of the microvasculature in the hippocampus that was accompanied by hippocampal atrophy. The vascular changes were not congophilic. Yet, despite the lack of congophilia, penetrating vessels at the cortical surface were often abnormal morphologically and microhemorrhages sometimes occurred. Altered immunostaining of blood vessels with basement membrane-associated antigens was an early feature of the microangiopathy and was associated with thickening of the vascular basal laminae and endothelial cell alterations that were visible ultrastructurally. Interestingly, although the FAD mutant transgene was expressed in neurons in both lines of mice, there was no detectable expression in vascular endothelial cells or glial cells. These studies thus have implications for the role of neuronal to vascular signaling in the pathogenesis of the vascular pathology associated with AD. PMID- 20008142 TI - MicroRNAs may mediate the down-regulation of neurokinin-1 receptor in chronic bladder pain syndrome. AB - Bladder pain syndrome (BPS) is a clinical syndrome of pelvic pain and urinary urgency-frequency in the absence of a specific cause. Investigating the expression levels of genes involved in the regulation of epithelial permeability, bladder contractility, and inflammation, we show that neurokinin (NK)1 and NK2 tachykinin receptors were significantly down-regulated in BPS patients. Tight junction proteins zona occludens-1, junctional adherins molecule -1, and occludin were similarly down-regulated, implicating increased urothelial permeability, whereas bradykinin B(1) receptor, cannabinoid receptor CB1 and muscarinic receptors M3-M5 were up-regulated. Using cell-based models, we show that prolonged exposure of NK1R to substance P caused a decrease of NK1R mRNA levels and a concomitant increase of regulatory micro(mi)RNAs miR-449b and miR-500. In the biopsies of BPS patients, the same miRNAs were significantly increased, suggesting that BPS promotes an attenuation of NK1R synthesis via activation of specific miRNAs. We confirm this hypothesis by identifying 31 differentially expressed miRNAs in BPS patients and demonstrate a direct correlation between miR 449b, miR-500, miR-328, and miR-320 and a down-regulation of NK1R mRNA and/or protein levels. Our findings further the knowledge of the molecular mechanisms of BPS, and have relevance for other clinical conditions involving the NK1 receptor. PMID- 20008143 TI - Interleukin-11 in endometrial adenocarcinoma is regulated by prostaglandin F2alpha-F-prostanoid receptor interaction via the calcium-calcineurin-nuclear factor of activated T cells pathway and negatively regulated by the regulator of calcineurin-1. AB - Interleukin-11 (IL-11) up-regulates the proliferative and invasive capacity of many cancers. Coexpression of glycoprotein 130 (GP130) and IL-11 receptor alpha (IL-11Ralpha) is necessary for high-affinity binding of IL-11 to IL-11Ralpha. This study investigated the expression of IL-11 and role of prostaglandin F(2alpha)-F-prostanoid receptor (FP receptor) signaling in the modulation of IL 11 expression in endometrial adenocarcinoma cells. Localization of IL-11, IL 11Ralpha, and GP130 expression was performed by immunohistochemistry. IL-11 and regulator of calcineurin 1 isoform 4 (RCAN1-4) mRNA and protein expression were determined by real-time RT-PCR and/or enzyme-linked immunosorbent assay/Western blot analysis using Ishikawa endometrial adenocarcinoma cells stably expressing the FP receptor (FPS cells) and endometrial adenocarcinoma explants. IL-11 mRNA expression was significantly elevated in endometrial adenocarcinoma samples compared with normal endometrium and increased with tumor grade. IL-11 protein expression localized with FP receptor, IL-11Ralpha, and GP130 in the neoplastic glandular epithelium of endometrial adenocarcinomas. Prostaglandin F(2alpha)-FP receptor signaling significantly elevated the expression of IL-11 mRNA and protein in a Gq-protein kinase C-calcium-calcineurin-nuclear factor of activated T cells-dependent manner in FPS cells. The calcineurin signaling pathway is known to be controlled by the RCAN (RCAN1-4). Indeed, RCAN1-4 expression was significantly elevated in well-differentiated endometrial adenocarcinoma compared with normal endometrium and was found to decrease with tumor grade and negatively regulate IL-11 expression in vitro. This study has highlighted a new mechanism regulating IL-11 expression in endometrial adenocarcinoma cells by the FP receptor via the calcium-calcineurin-nuclear factor of activated T cells pathway. PMID- 20008144 TI - CrkII transgene induces atypical mammary gland development and tumorigenesis. AB - The v-Crk protein was originally isolated as the oncogene fusion product of the CT10 chicken retrovirus. Cellular homologues of v-Crk include Crk, which encodes two alternatively spliced proteins (CrkI and CrkII), and CrkL. Though CrkI/II proteins are elevated in several types of cancer, including breast, the question of whether these Crk adaptor proteins can promote breast cancer has not been addressed. We created a transgenic mouse model that allows the expression of CrkII through the hormonally responsive mouse mammary tumor virus promoter. During puberty, transgenic mice were found to have delayed ductal outgrowth, characterized by increased collagen surrounding the terminal end buds. In post pubertal mice, precocious ductal branching was observed and associated with increased proliferation. Focal mammary tumors appeared in a subset of animals, with a latency of approximately 15 months. Mouse mammary tumor virus/CrkII tumors showed high levels of Crk protein as well as various cytokeratin markers characteristic of their respective tumor pathologies. This study demonstrates that the precise expression of CrkII is critical for integrating signals for ductal outgrowth and branching morphogenesis during mammary gland development. Furthermore, this study provides evidence for a potential role of CrkII in integrating signals for breast cancer progression in vivo, which has important implications for elevated CrkII observed in human cancer. PMID- 20008145 TI - Enoxaparin improves the course of dextran sodium sulfate-induced colitis in syndecan-1-deficient mice. AB - Syndecan-1 (Sdc1) plays a major role in wound healing and modulates inflammatory responses. Sdc1 expression is reduced in lesions of patients with ulcerative colitis. The aim of this study was to investigate the role of Sdc1 in murine dextran sodium sulfate (DSS)-induced colitis. DSS colitis was induced in Sdc1 deficient (knockout (KO)) and wild-type mice by oral administration of 3% DSS. KO mice exhibited a significantly increased lethality as compared with wild-type controls (61 versus 5%, P < 0.05). Impaired mucosal healing and prolonged recruitment of inflammatory cells in KO mice were accompanied by significant up regulation of tumor necrosis factor-alpha, CC chemokine ligand 3/macrophage inflammatory protein-1alpha, and vascular cell adhesion molecule-1, as determined by histological correlation between 0 and 15 days after colitis induction, TaqMan low-density array analysis, and quantitative real-time PCR. Treatment from days 7 through 14 with enoxaparin, a functional analogue of the Sdc1 heparan sulfate chains, significantly reduced lethality of KO mice due to DSS-induced colitis, which was correlated with improved mucosal healing. In vitro, Sdc1-deficient polymorphonuclear cells displayed increased adhesion to endothelial cells and intercellular adhesion molecule-1, and enoxaparin reverted adhesion to wild-type levels. Small interfering RNA-mediated knockdown of Sdc1 expression resulted in reduced basic fibroblast growth factor-mediated mitogen-activated protein kinase signaling and reduced Caco-2 cell proliferation. We conclude that Sdc1 has a protective effect during experimental colitis. The modification of missing Sdc1 function by heparin analogues may emerge as a promising anti-inflammatory approach. PMID- 20008146 TI - alpha2-antiplasmin is associated with the progression of fibrosis. AB - Systemic sclerosis results in tissue fibrosis due to the activation of fibroblasts and the ensuing overproduction of the extracellular matrix. We previously reported that the absence of alpha2-antiplasmin (alpha2AP) attenuated the process of dermal fibrosis; however, the detailed mechanism of how alpha2AP affects the progression of fibrosis remained unclear. The goal of the present study was to examine the role of alpha2AP in fibrotic change. We observed significantly higher levels of alpha2AP expression in the skin of bleomycin injected systemic sclerosis model mice in comparison with the levels seen in control mice. We also demonstrated that alpha2AP induced myofibroblast differentiation, and the absence of alpha2AP attenuated the induction of myofibroblast differentiation. Moreover, we found that connective tissue growth factor induced the expression of alpha2AP through both the extracellular signal regulated kinase 1/2 (ERK1/2) and c-Jun N-terminal kinase (JNK) pathways in fibroblasts. Interestingly, alpha2AP also induced transforming growth factor-beta expression through the same pathways, and the inhibition of ERK1/2 and JNK slowed the progression of bleomycin-induced fibrosis. Our findings suggest that alpha2AP is associated with the progression of fibrosis, and regulation of alpha2AP expression by the ERK1/2 and JNK pathways may be an effective antifibrotic therapy for the treatment of systemic sclerosis. PMID- 20008147 TI - Tissue inhibitor of metalloproteinases 3 regulates resolution of inflammation following acute lung injury. AB - Tissue inhibitor of metalloproteinases 3 (TIMP3) inhibits not only matrix metalloproteinases but also a disintegrin and metalloproteinase domain family members and thus contributes to controlling diverse processes mediated by proteolysis. We used Timp3(-/-) mice to assess the role of this inhibitor in acute lung injury. After bleomycin-induced injury, inflammation, as indicated by the influx of neutrophils in bronchoalveolar lavage (BAL), peaked at 7 days post injury in the wild-type mice and began to wane thereafter; however, in Timp3(-/-) mice, inflammation persisted up to 28 days. Furthermore, although the level of chemokines in BAL and lung homogenate was similar in both genotypes, BAL from Timp3(-/-) mice 7, 14, and 28 days post-injury had increased neutrophil chemotactic activity compared with wild-type BAL. At day 14, a higher percentage of apoptotic neutrophils were present in wild-type mice compared with Timp3(-/-) mice, further suggesting that TIMP3 constrains continued neutrophil influx. In addition, total matrix metalloproteinase activity was increased in lungs from Timp3(-/-) mice, and treatment of mice with a synthetic inhibitor of metalloproteinases rescued the enhanced neutrophilia phenotype. These data demonstrate that TIMP3 regulates neutrophil influx in the lung following injury through its ability to inhibit metalloproteinase activity and indicates that TIMP3 functions to promote the resolution of inflammation in the lung. PMID- 20008148 TI - Venular basement membranes ubiquitously express matrix protein low-expression regions: characterization in multiple tissues and remodeling during inflammation. AB - The venular basement membrane plays a critical role in maintaining the integrity of blood vessels and through its dense and highly organized network of matrix proteins also acts as a formidable barrier to macromolecules and emigrating leukocytes. Leukocytes can however penetrate the venular basement membrane at sites of inflammation, though the associated in vivo mechanisms are poorly understood. Using whole mount immunostained tissues and confocal microscopy, we demonstrate that the venular basement membrane of multiple organs expresses regions of low matrix protein (laminin-511 and type IV collagen) deposition that have been termed low-expression regions (LERs). In the multiple tissues analyzed (eg, cremaster muscle, skin, mesenteric tissue), LERs were directly aligned with gaps between adjacent pericytes and were more prevalent in small venules. As predicted by their permissive nature, LERs acted as "gates" for transmigrating neutrophils in all inflammatory reactions investigated (elicited by leukotriene B(4) [LTB(4)], CXCL1, tumor necrosis factor [TNF]alpha, endotoxin, and ischemia/reperfusion [I/R] injury), and this response was associated with an enhancement of the size of laminin-511 and type IV collagen LERs. Transmigrated neutrophils stained positively for laminins but not type IV collagen, suggesting that different mechanisms exist in remodeling of different basement membrane networks. Collectively the findings provide further insight into characteristics of specialized regions within venular basement membranes that are preferentially used and remodeled by transmigrating neutrophils. PMID- 20008151 TI - Views on treatment necessity, harm, and benefits in patients using statins. AB - BACKGROUND: Patients with dyslipidemia and high overall risk of ischemic heart disease are those most likely to benefit from treatment with statins. The objective of this study was to investigate patients' views of the necessity, harm, and expected benefits of their statin treatment, as well as factors associated with these treatment beliefs. One main objective was to investigate whether cardiovascular risk level and previous coronary heart disease affect the way patients view these different aspects of their medication. METHODS: A total of 829 statin users were recruited while visiting a pharmacy to collect their statin medication (response rate, 69.4%). Patients returned a questionnaire assessing their medical history, concurrent risk status, social demographic factors, as well as their views and expectations regarding their statin treatment. RESULTS: Previous ischemic heart disease, or high risk of such disease, was not associated with a more favorable notion on statin treatment. Having an internal health locus of control as well as being satisfied with the physician's treatment explanation were factors associated with more positive views on expected treatment effect. The views on necessity, harm, and treatment benefit were closely associated with each other. CONCLUSION: The main findings in this study suggest that cardiovascular risk level and previous coronary heart disease are not associated with the way patients value potential benefits of statin treatment. Patients' views of the treatment's necessity, harm, and benefits do not seem to be independent dimensions of patient beliefs but rather represent one overall question: is this good for me? PMID- 20008150 TI - Dendritic cells express hematopoietic prostaglandin D synthase and function as a source of prostaglandin D2 in the skin. AB - Prostaglandin D2 (PGD2), an arachidonic acid metabolite, has been implicated in allergic responses. A major source of PGD2 in the skin is mast cells that express hematopoietic PGD synthase (H-PGDS). In this study, we show the expression of H PGDS in human dendritic cells (DCs) and the regulatory mechanisms by which DCs produce PGD2. We detected H-PGDS in epidermal Langerhans cells, dermal DCs, plasmacytoid DCs, and myeloid DCs. Monocyte-derived DCs rapidly secreted PGD2 when stimulated with the calcium ionophore A23187. More importantly, pretreatment of monocyte-derived DCs with PMA (phorbol 12-myrisate 13-acetate) synergistically enhanced the rapid PGD2 secretion induced by A23187, whereas PMA alone did not induce PGD2 secretion. Lipopolysaccharide (LPS) reduced H-PGDS expression, but interferon-gamma followed by LPS induced significant PGD2 production in a delayed time course at 6 hours. This effect was associated with inhibition of LPS-induced H-PGDS reduction. Interestingly, an irritant compound, SDS, also induced a rapid PGD2 release. PGD2 synergistically enhanced CCL22/macrophage-derived chemokine synthesis in interferon-gamma-treated human keratinocytes. In addition, bone marrow-derived DCs from wild-type mice stimulated lymph node cells to produce higher amounts of interleukin-17 than did DCs from mice lacking the H-PGDS gene. Thus, DCs could be an important source of skin PGD2 and may mediate or regulate skin inflammation by releasing PGD2 in response to various stimuli, contributing to the innate and/or acquired immune responses. PMID- 20008149 TI - Endogenous LXA4 circuits are determinants of pathological angiogenesis in response to chronic injury. AB - Inflammation and angiogenesis are intimately linked, and their dysregulation leads to pathological angiogenesis in human diseases. 15-lipoxygenase (15-LOX) and lipoxin A(4) receptors (ALX) constitute a LXA(4) circuit that is a key feature of inflammatory resolution. LXA(4) analogs have been shown to regulate vascular endothelial growth factor (VEGF)-A-induced angiogenic response in vitro. 15-LOX and ALX are highly expressed in the avascular and immune-privileged cornea. However, the role of this endogenous LXA(4) circuit in pathological neovascularization has not been determined. We report that suture-induced chronic injury in the cornea triggered polymorphonuclear leukocytes (PMN) infiltration, pathological neovascularization, and up-regulation of mediators of inflammatory angiogenesis, namely VEGF-A and the VEGF-3 receptor (FLT4). Up-regulation of the VEGF circuit and neovascularization correlated with selective changes in both 15 LOX (Alox15) and ALX (Fpr-rs2) expression and a temporally defined increase in basal 15-LOX activity. More importantly, genetic deletion of 15-LOX or 5-LOX, key and obligatory enzymes in the formation of LXA(4), respectively, led to exacerbated inflammatory neovascularization coincident with increased VEGF-A and FLT4 expression. Direct topical treatment with LXA(4), but not its metabolic precursor 15-hydroxyeicosatetraenoic acid, reduced expression of VEGF-A and FLT4 and inflammatory angiogenesis and rescued 15-LOX knockout mice from exacerbated angiogenesis. In summary, our findings and the prominent expression of 15-LOX and ALX in epithelial cells and macrophages place the LXA(4) circuit as an endogenous regulator of pathological angiogenesis. PMID- 20008152 TI - The role of reported tobacco-specific media exposure on adult attitudes towards proposed policies to limit the portrayal of smoking in movies. AB - OBJECTIVE: To assess the relative, independent contribution of reported tobacco specific media exposure (pro-tobacco advertising, anti-tobacco advertising, and news coverage of tobacco issues) to US adults' support for policy efforts that aim to regulate the portrayal of smoking in movies. METHODS: Using the American Legacy Foundation's 2003 American Smoking and Health Survey (ASHES-2), multivariable logistic regression was used to model the predicted probability that US adults support movie-specific tobacco control policies, by reported exposure to tobacco-specific media messages, controlling for smoking status, education, income, race/ethnicity, age, sex, knowledge of the negative effects of tobacco and state. RESULTS: Across most outcome variables under study, findings reveal that reported exposure to tobacco-specific media messages is associated with adult attitudes towards movie-specific policy measures. Most exposure to tobacco information in the media (with the exception of pro-tobacco advertising on the internet) contributes independently to the prediction of adult support for movie-specific policies. The direction of effect follows an expected pattern, with reported exposure to anti-tobacco advertising and news coverage of tobacco predicting supportive attitudes towards movie policies, and reported exposure to pro-tobacco advertising lessening support for some movie policies, though the medium of delivery makes a difference. CONCLUSION: Media campaigns to prevent tobacco use and exposure to secondhand smoke have had value beyond the intended impact of single-issue campaigns; exposure to anti-tobacco campaigns and public dialogue about the dangers of tobacco seem also to be associated with shaping perceptions of the social world related to norms about tobacco, and ideas about regulating the portrayal of smoking in movies. PMID- 20008153 TI - Association between tobacco vendor non-compliance with youth access and point of sale restrictions. PMID- 20008154 TI - A cross-sectional study on levels of second-hand smoke in restaurants and bars in five cities in China. AB - OBJECTIVES: To assess indoor second-hand smoke (SHS) exposure in restaurants and bars via PM(2.5) (fine particles 2.5 MUm in diameter and smaller) level measurements in five cities in China. METHODS: The study was conducted from July to September in 2007 in Beijing, Xi'an, Wuhan, Kunming and Guiyang. Portable aerosol monitors were used to measure PM(2.5) concentrations in 404 restaurants and bars. The occupant density and the active smoker density were calculated for each venue sampled. RESULTS: Among the 404 surveyed venues, 23 had complete smoking bans, 9 had partial smoking bans and 313 (77.5%) were observed to have allowed smoking during sampling. The geometric mean of indoor PM(2.5) levels in venues with smoking observed was 208 MUg/m(3) and 99 MUg/m(3) in venues without observed smoking. When outdoor PM(2.5) levels were adjusted, indoor PM(2.5) levels in venues with smoking observed were consistently significantly higher than in venues without smoking observed (F=80.49, p<0.001). Indoor PM(2.5) levels were positively correlated with outdoor PM(2.5) levels (partial rho=0.37 p<0.001) and active smoker density (partial rho=0.34, p<0.001). CONCLUSIONS: Consistent with findings in other countries, PM(2.5) levels in smoking places are significantly higher than those in smoke-free places and are strongly related to the number and density of active smokers. These findings document the high levels of SHS in hospitality venues in China and point to the urgent need for comprehensive smoke-free laws in China to protect the public from SHS hazards, as called for in Article 8 of the Framework Convention on Tobacco Control, which was ratified by China in 2005. PMID- 20008155 TI - Exposure to movie smoking, antismoking ads and smoking intensity: an experimental study with a factorial design. AB - BACKGROUND: This study examines whether smoking portrayal in movies or antismoking advertisements affect smoking intensity among young adults. METHODS: We conducted an experimental study in which 84 smokers were randomly assigned using a two (no-smoking versus smoking portrayal in the movie) by three (two prosocial ads, two antismoking ads or one of each) factorial design. Participants viewed a 60-minute movie with two commercial breaks and afterwards completed a questionnaire. Smoking during the session was allowed and observed. RESULTS: Exposure to the movie with smoking had no effect on smoking intensity. Those who viewed two antismoking ads had significantly lower smoking intensity compared with those who viewed two prosocial ads. There was no interaction between movie smoking and antismoking ads. Baseline CO (carbon monoxide) level had the largest effect on smoking intensity. CONCLUSION: These findings provide further evidence to support antismoking ads placed with movies because of their possible effect on young adult smoking behaviour. However, caution is warranted, because nicotine dependence appears to be the primary predictor of smoking intensity among young adult smokers in this study. PMID- 20008156 TI - Regional differences in awareness of tobacco advertising and promotion in China: findings from the ITC China Survey. AB - OBJECTIVE: To examine whether levels of, and factors related to, awareness of tobacco advertising and promotion differ across six cities in China. METHODS: Data from wave 1 of the International Tobacco Control (ITC) China Survey (April to August 2006) were analysed. The ITC China Survey employed a multistage sampling design in Beijing, Shenyang, Shanghai, Changsha, Guangzhou and Yinchuan. Face-to-face interviews were conducted with a total of 4763 smokers and 1259 non smokers. Multivariate logistic regression models were used to identify factors associated with awareness of tobacco advertising and promotion. RESULTS: The overall levels of noticing advertisements varied considerably by city. Cities reporting lower levels of advertising tended to report higher levels of point of sale activity. Noticing tobacco industry promotions was associated with more positive attitudes to tobacco companies. CONCLUSION: The awareness of tobacco advertising and promotional activities was not homogeneous across the six Chinese cities, suggesting variations in the tobacco industry's activities and the diversity of implementing a central set of laws to restrict tobacco promotion. This study clearly demonstrates the need to work with the implementation agencies if national laws are to be properly enforced. PMID- 20008157 TI - Patterns of dual use of cigarettes and smokeless tobacco among US males: findings from national surveys. AB - BACKGROUND: In the USA, consumption of moist snuff continues to increase and cigarette manufacturers now control nearly its entire market. Manufacturers have developed new products that represent cigarette brand extension and in test marketing are promoting dual use of cigarettes and snuff. This study examined patterns of concurrent use of smokeless tobacco (ST) and cigarettes among young people and adults in the USA just before cigarette companies' control of the nation's ST market. METHODS: Data were drawn from four US nationally representative surveys. Stratified analyses applied sampling weights and accounted for the complex sample designs. RESULTS: Cigarette smoking was substantially more prevalent among young males who used ST than among those who did not. Among adult males, those who smoked daily were less likely than others to have used snuff every day. Men who used moist snuff daily had the lowest prevalence of daily smoking, but the prevalence of daily smoking was relatively high among men who used moist snuff less than daily. Unsuccessful past-year attempts by daily smokers to quit smoking were more prevalent among non-daily snuff users (41.2%) than among those who had never used snuff (29.6%). CONCLUSIONS: Although dual daily use of ST and cigarettes is relatively uncommon in the USA, concurrent ST use is more common among adolescent and young adult male smokers than among more mature tobacco users. Among adult males, daily smoking predominates and non-daily ST use is very strongly associated with current smoking. Adult male smokers who also use ST daily tend to have relatively high levels of serum cotinine and high prevalence of a major indicator for tobacco dependence. PMID- 20008158 TI - The role of taxation in tobacco control and its potential economic impact in China. AB - OBJECTIVES: To identify key economic issues involved in raising the tobacco tax and to recommend possible options for tobacco tax reform in China. METHODS: Estimated price elasticities of the demand for cigarettes, prevalence data and epidemiology are used to estimate the impact of a tobacco tax increase on cigarette consumption, government tax revenue, lives saved, employment and revenue loss in the cigarette industry and tobacco farming. RESULTS: The recent Chinese tax adjustment, if passed along to the retail price, would reduce the number of smokers by 630,000 saving 210,000 lives, at a price elasticity of 0.15. A tax increase of 1 RMB (or US$0.13) per pack of cigarettes would increase the Chinese government's tax revenue by 129 billion RMB (US 17.2 billion), decrease consumption by 3.0 billion packs of cigarettes, reduce the number of smokers by 3.42 million and save 1.14 million lives. CONCLUSION: The empirical economic analysis and tax simulation results clearly indicate that increasing the tobacco tax in China is the most cost-effective instrument for tobacco control. PMID- 20008159 TI - Pattern of tobacco use among the Iranian adult population: results of the national Survey of Risk Factors of Non-Communicable Diseases (SuRFNCD-2007). AB - BACKGROUND: Previous studies report on smoking in Iran but recent national data on tobacco use (including cigarette, water-pipe and pipe) have not been reported. METHODS: In 2007, 5287 Iranians aged 15-64 years were sampled from all provinces as part of a national cross-sectional survey of non-communicable disease (NCD) risk factors. Data were collected using the standardised stepwise protocol for NCD risk factor surveillance of the World Health Organization. Use of tobacco products was calculated as the sum of smoking cigarettes/cigars (smoking currently or daily any amount of factory/hand-made cigarettes or cigars), pipes (daily) and water pipes (daily). RESULTS: Total current and daily tobacco use was 14.8% (burden 7.3 million) and 13.7% (burden 6.7 million) when extrapolated to the Iranian population aged 15-64. The prevalence of current and daily cigarette smoking was 12.5% (6.1 million; 23.4% males and 1.4% females) and 11.3% (5.6 million; 21.4 males and 1.4 females); former smokers comprised 1.7 million or 3.4% of the Iranian population (6.2% males and 0.6% females; mean cessation age 34.1). The mean age of starting to smoke was 20.5 years (24.2 males and 20.4 females). The prevalence of water-pipe smoking was 2.7% (burden 1.3 million; 3.5% males and 1.9% females). Water-pipe smokers used the water-pipe on average 3.5 times a day (2.8 males and 4.5 females). CONCLUSION: The prevalence of tobacco use has not escalated over the past two decades. Nonetheless, the burden is high and therefore warrants preventive public health policies. PMID- 20008160 TI - The effects of smoking and smoking cessation on mortality from cardiovascular disease among Japanese: pooled analysis of three large-scale cohort studies in Japan. AB - OBJECTIVES: To estimate the gender-specific risk of mortality from cardiovascular disease according to smoking status and time since smoking cessation among former smokers in Japan. DESIGN: Prospective study. SETTING: 140,026 males and 156,810 females aged 40-79 years who participated in one of three cohort studies conducted in Japan between 1980 and 1990. OUTCOME: The gender-specific hazard ratios (HRs) for cardiovascular disease mortality were calculated after adjustment for age and cohort. RESULTS: The age-adjusted and cohort-adjusted HRs for current smokers compared with lifelong non-smokers were 1.51 (95% CI 1.38 to 1.64) for total cardiovascular diseases, 2.19 (95% CI 1.79 to 2.67) for coronary heart disease and 1.24 (95% CI 1.10 to 1.41) for total stroke in males, and were 1.85 (95% CI 1.65 to 2.06), 2.84 (95% CI 2.24 to 3.60) and 1.70 (95% CI 1.44 to 2.01), respectively, in females. The age-adjusted and cohort-adjusted HRs for former smokers compared with current smokers according to the time period since smoking cessation decreased by approximately 5 years after smoking cessation and reached the same level as lifelong non-smokers approximately 10 years after smoking cessation among both males and females. CONCLUSIONS: The present study confirmed the association between smoking and mortality from cardiovascular disease in both males and females. Smoking cessation is a crucial preventive measure against death from cardiovascular disease. PMID- 20008161 TI - Investigating the spatial variability in incidence of coronary heart disease in the Gazel cohort: the impact of area socioeconomic position and mediating role of risk factors. AB - STUDY OBJECTIVE: The aim of the study was to improve understanding of the relationships between contextual socioeconomic characteristics and coronary heart disease (CHD) incidence in France. Several authors have suggested that CHD risk factors (diabetes, hypertension, cholesterol, overweight, tobacco consumption) may partly mediate associations between socioeconomic environmental variables and CHD. Studies have assessed the overall mediating role of CHD risk factors, but have never investigated the specific mediating role of each risk factor, not allowing their specific contribution to the area socioeconomic position-CHD association to be disentangled. DESIGN: After assessing geographical variations in CHD incidence and socioeconomic environmental effects on CHD using a multilevel Cox model, the extent to which this contextual effect was mediated by each of the CHD risk factors was assessed. PARTICIPANTS: Data of the French GAZEL cohort (n=19,808) were used. MAIN RESULTS: After adjustment for several individual socioeconomic indicators, it was found, in men from highly urbanised environments, that CHD incidence increased with decreasing socioeconomic position of the residential environment. After individual-level adjustment, a higher risk of obesity, smoking and cholesterol was observed in the most deprived residential environments. When risk factors were introduced into the model, a modest decrease was observed in the magnitude of the association between the socioeconomic contextual variable and CHD. Risk factors that contributed most to the decrease of the association were smoking and cholesterol. CONCLUSIONS: Classic risk factors, although some of them more than others, mediated a modest part of the association between area socioeconomic position and CHD. PMID- 20008162 TI - Minutes, MET minutes, and METs: unpacking socio-economic gradients in physical activity in adolescents. AB - BACKGROUND: There is conflicting evidence regarding the relationship between socio-economic position (SEP) and physical activity in adolescents. The aim of this study was to investigate associations between SEP and characteristics of physical activity in Australian adolescents using a high-resolution use-of-time tool. METHOD: Use-of-time and pedometer data were collected on a random sample of 2071 9-16-year-old Australian children. Use-of-time was recorded using a computerised 24-h use-of-time recall, the Multimedia Activity Recall for Children and Adults. Reported household income was used as a marker of SEP. RESULTS: There were no differences in self-reported minutes of moderate to vigorous physical activity (MVPA) across the income bands and only very small differences in the number of daily steps. However, the mix of MVPA components varied across income bands, with adolescents from low-SEP families experiencing less sport but more active transport. Because the mean rate of energy expenditure was greater in sport than in other forms of MVPA (play, active transport or chores), there were significant differences in MVPA-related and total daily energy expenditure across income bands, with the lower bands having significantly lower values. Differences in total daily energy expenditure were almost entirely explained by differences in energy expenditure associated with sport. CONCLUSION: Physical activity patterns vary across SEP bands in Australian adolescents, with sport being the major locus of differences. Instruments which do not account for the energy costs of various activities may fail to detect important relationships. PMID- 20008163 TI - Iodine intake in a population of pregnant women: INMA mother and child cohort study, Spain. AB - BACKGROUND: Monitoring iodine status during pregnancy is essential to prevent iodine-related disorders. The objectives of this study are to estimate iodine intake and excretion, to assess their association and to evaluate the compliance of the recommendations in a multicentre cohort of pregnant women. METHODS: Cross sectional data on maternal iodine nutritional status, compiled between weeks 8 and 22 of gestation in three Spanish areas (Valencia, Gipuzkoa and Sabadell), were analysed. Information on iodine intake from diet, salt and supplements was estimated through questionnaires. Spot urine samples were analysed for urinary iodine concentration (UIC). Tobit regression analysis was used to assess the association between iodine intake and UIC. RESULTS: 1522 women were included in the study. Median UIC was 134 (IQR 80-218) MUg/l in Valencia, 168 (IQR 108-272) MUg/l in Gipuzkoa and 94 (IQR 57-151) MUg/l in Sabadell. 48.9% of Valencian women consumed iodine supplements, 93.3% in Gipuzkoa and 11.0% in Sabadell. Prevalence of iodised salt consumption was 50.5% in the whole sample. UIC was associated with intake of supplements, iodised salt, dietary iodine and water. UIC levels were lower than expected according to the estimated iodine intake. CONCLUSION: Median UIC reflected iodine deficiency according to WHO reference levels, except in Gipuzkoa where supplements are widely consumed. It is necessary to strengthen iodised salt consumption since it is already far from the objective proposed of coverage of 90% of households. More data would be valuable to assess the correspondence between iodine intake and excretion during pregnancy. PMID- 20008164 TI - Myoferlin is required for insulin-like growth factor response and muscle growth. AB - Insulin-like growth factor (IGF) is a potent stimulus of muscle growth. Myoferlin is a membrane-associated protein important for muscle development and regeneration. Myoferlin-null mice have smaller muscles and defective myoblast fusion. To understand the mechanism by which myoferlin loss retards muscle growth, we found that myoferlin-null muscle does not respond to IGF1. In vivo after IGF1 infusion, control muscle increased myofiber diameter by 25%, but myoferlin-null muscle was unresponsive. Myoblasts cultured from myoferlin-null muscle and treated with IGF1 also failed to show the expected increase in fusion to multinucleate myotubes. The IGF1 receptor colocalized with myoferlin at sites of myoblast fusion. The lack of IGF1 responsiveness in myoferlin-null myoblasts was linked directly to IGF1 receptor mistrafficking as well as decreased IGF1 signaling. In myoferlin-null myoblasts, the IGF1 receptor accumulated into large vesicular structures. These vesicles colocalized with a marker of late endosomes/lysosomes, LAMP2, specifying redirection from a recycling to a degradative pathway. Furthermore, ultrastructural analysis showed a marked increase in vacuoles in myoferlin-null muscle. These data demonstrate that IGF1 receptor recycling is required for normal myogenesis and that myoferlin is a critical mediator of postnatal muscle growth mediated by IGF1.-Demonbreun, A. R., Posey, A. D., Heretis, K., Swaggart, K. A., Earley, J. U., Pytel, P., McNally, E. M. Myoferlin is required for insulin-like growth factor response and muscle growth. PMID- 20008165 TI - Key process conditions for production of C(4) dicarboxylic acids in bioreactor batch cultures of an engineered Saccharomyces cerevisiae strain. AB - A recent effort to improve malic acid production by Saccharomyces cerevisiae by means of metabolic engineering resulted in a strain that produced up to 59 g liter(-1) of malate at a yield of 0.42 mol (mol glucose)(-1) in calcium carbonate buffered shake flask cultures. With shake flasks, process parameters that are important for scaling up this process cannot be controlled independently. In this study, growth and product formation by the engineered strain were studied in bioreactors in order to separately analyze the effects of pH, calcium, and carbon dioxide and oxygen availability. A near-neutral pH, which in shake flasks was achieved by adding CaCO(3), was required for efficient C(4) dicarboxylic acid production. Increased calcium concentrations, a side effect of CaCO(3) dissolution, had a small positive effect on malate formation. Carbon dioxide enrichment of the sparging gas (up to 15% [vol/vol]) improved production of both malate and succinate. At higher concentrations, succinate titers further increased, reaching 0.29 mol (mol glucose)(-1), whereas malate formation strongly decreased. Although fully aerobic conditions could be achieved, it was found that moderate oxygen limitation benefitted malate production. In conclusion, malic acid production with the engineered S. cerevisiae strain could be successfully transferred from shake flasks to 1-liter batch bioreactors by simultaneous optimization of four process parameters (pH and concentrations of CO(2), calcium, and O(2)). Under optimized conditions, a malate yield of 0.48 +/- 0.01 mol (mol glucose)(-1) was obtained in bioreactors, a 19% increase over yields in shake flask experiments. PMID- 20008166 TI - Application of a short, disordered N-terminal flagellin segment, a fully functional flagellar type III export signal, to expression of secreted proteins. AB - Recently, we have demonstrated that the 26-47 segment of Salmonella enterica serovar Typhimurium flagellin is capable of mediating flagellar export. In order to reveal whether other parts of the N-terminal region have any significant influence on secretion, a series of plasmids were constructed containing the lac promoter followed by the 26-47, 2-65, or 2-192 portion of Salmonella flagellin, to which various heterologous proteins of different size were fused (18 constructs overall). Essentially, all three segments could drive protein export; however, the nature of the attached polypeptide also had a significant effect on secretion efficiency. When low export efficiency was observed, it was mainly caused by inclusion body formation. Our data provide strong support for the idea that a short segment within the disordered N-terminal region of axial proteins is recognized by the flagellar type III export machinery. The 26-47 segment of flagellin contains all of the necessary information to direct translocation of attached polypeptide chains. This short (positions 26 to 47) flagellin segment attached to recombinant proteins can be used for secreted protein expression. Certain fusion proteins that are easily degraded within the cells were found to be intact in the medium, implying a potential application of this expression system for proteins with high proteolytic susceptibility. PMID- 20008167 TI - Stability of a Spodoptera frugiperda nucleopolyhedrovirus deletion recombinant during serial passage in insects. AB - The stabilities of the Spodoptera frugiperda multiple nucleopolyhedrovirus (SfMNPV) complete genome bacmid (Sfbac) and a deletion recombinant (Sf29null) in which the Sf29 gene was replaced by a kanamycin resistance cassette were determined during sequential rounds of per os infection in insect larvae. The Sf29 gene is a viral factor that determines the number of virions in occlusion bodies (OBs). The Sf29null bacmid virus was able to recover the Sf29 gene during passage. After the third passage (P3) of Sf29null bacmid OBs, the population was observed to reach an equilibrium involving a mixture of those with a kanamycin resistance cassette and those with the Sf29 gene. The biological activity of Sf29null bacmid OBs at P3 was similar to that of Sfbac OBs. The recovered gene in the Sf29null virus was 98 to 100% homologous to the Sf29 genes of different SfMNPV genotypes. Reverse transcription-PCR analysis of uninoculated S. frugiperda larvae confirmed the expression of the SfMNPV ie-0 and Sf29 genes, indicating that the insect colony harbors a covert SfMNPV infection. Additionally, the nonessential bacterial artificial chromosome vector was spontaneously deleted from both viral genomes upon passage in insects. PMID- 20008168 TI - Reductive debromination of polybrominated diphenyl ethers by anaerobic bacteria from soils and sediments. AB - Polybrominated diphenyl ethers (PBDEs) have attracted attention recently due to their proven adverse effects on animals and their increasing concentrations in various environmental media and biota. To gain insight into the fate of PBDEs, microcosms established with soils and sediments from 28 locations were investigated to determine their debromination potential with an octa-brominated diphenyl ether (octa-BDE) mixture consisting of hexa- to nona-BDEs. Debromination occurred in microcosms containing samples from 20 of the 28 locations when they were spiked with octa-BDE dissolved in the solvent trichloroethene (TCE), which is a potential cosubstrate for stimulating PBDE debromination, and in microcosms containing samples from 11 of the 28 locations when they were spiked with octa BDE dissolved in nonane. Debromination products ranging from hexa- to mono-BDEs were generated within 2 months. Notably, the toxic tetra-BDEs accounted for 50% of the total product. In sediment-free culture C-N-7* amended with the octa-BDE mixture and nonane (containing 45 nM nona-BDE, 181 nM octa-BDEs, 294 nM hepta BDE, and 19 nM hexa-BDE) there was extensive debromination of the parent compounds, which produced hexa-BDE (56 nM), penta-BDEs (124 nM), and tetra-BDEs (150 nM) within 42 days, possibly by a metabolic process. A 16S rRNA gene-based analysis revealed that Dehalococcoides species were present in 11 of 14 active microcosms. However, unknown debrominating species in some of the microcosms debrominated the octa-BDE mixture in the absence of other added halogenated electron acceptors (such as TCE). These findings provide information that is useful for assessing microbial reductive debromination of higher brominated PBDEs to less-brominated congeners, a possible source of the more toxic congeners (e.g., penta- and tetra-BDEs) detected in the environment. PMID- 20008169 TI - Physiological versatility of the extremely thermoacidophilic archaeon Metallosphaera sedula supported by transcriptomic analysis of heterotrophic, autotrophic, and mixotrophic growth. AB - Comparative transcriptomic analysis of autotrophic, heterotrophic, and mixotrophic growth of the archaeon Metallosphaera sedula (70 degrees C, pH 2.0) revealed candidates for yet-to-be-confirmed components of the 3 hydroxypropionate/4-hydroxybutyrate pathway and implicated a membrane-bound hydrogenase (Msed_0944-Msed_0946) for growth on H(2). Routes for generation of ATP and reducing equivalents were also identified. PMID- 20008170 TI - Correlation of Dehalococcoides 16S rRNA and chloroethene-reductive dehalogenase genes with geochemical conditions in chloroethene-contaminated groundwater. AB - Quantitative analysis of genes that code for Dehalococcoides 16S rRNA and chloroethene-reductive dehalogenases TceA, VcrA, and BvcA was done on groundwater sampled from 150 monitoring wells spread over 11 chlorinated ethene polluted European locations. Redundancy analysis was used to relate molecular data to geochemical conditions. Dehalococcoides 16S rRNA- and vinyl chloride (VC) reductase genes were present at all tested locations in concentrations up to 10(6) gene copies per ml of groundwater. However, differences between and also within locations were observed. Variation in Dehalococcoides 16S rRNA gene copy numbers were most strongly correlated to dissolved organic carbon concentration in groundwater and to conditions appropriate for biodegradation of chlorinated ethenes (U.S. Environmental Protection Agency score). In contrast, vcrA gene copy numbers correlated most significantly to VC and chlorinated ethene concentrations. Interestingly, bvcA and especially tceA were more correlated with oxidizing conditions. In groundwater microcosms, dechlorination of 1 mM VC was correlated to an increase of vcrA and/or bvcA gene copies by 2 to 4 orders of magnitude. Interestingly, in 34% of the monitoring wells and in 40% of the active microcosms, the amount of individual VC-reductase gene copies exceeded that of Dehalococcoides 16S rRNA gene copies. It is concluded that the geographical distribution of the genes was not homogeneous, depending on the geochemical conditions, whereby tceA and bvcA correlated to more oxidized conditions than Dehalococcoides 16S rRNA and vcrA. Because the variation in VC-reductase gene numbers was not directly correlated to variation in Dehalococcoides spp., VC reductase genes are better monitoring parameters for VC dechlorination capacity than Dehalococcoides spp. PMID- 20008171 TI - Highly diverse cyanobactins in strains of the genus Anabaena. AB - Cyanobactins are small, cyclic peptides recently found in cyanobacteria. They are formed through proteolytic cleavage and posttranslational modification of short precursor proteins and exhibit antitumor, cytotoxic, or multi-drug-reversing activities. Using genome project data, bioinformatics, stable isotope labeling, and mass spectrometry, we discovered novel cyclic peptides, anacyclamides, in 27 Anabaena strains. The lengths of the anacylamides varied greatly, from 7 to 20 amino acids. Pronounced sequence variation was also detected, and only one amino acid, proline, was present in all anacyclamides. The anacyclamides identified included unmodified proteinogenic or prenylated amino acids. We identified an 11 kb gene cluster in the genome of Anabaena sp. 90, and heterologous expression in Escherichia coli confirmed that this cluster was responsible for anacyclamide production. The discovery of anacyclamides greatly increases the structural diversity of cyanobactins. PMID- 20008172 TI - Diversity and evolution of the phenazine biosynthesis pathway. AB - Phenazines are versatile secondary metabolites of bacterial origin that function in biological control of plant pathogens and contribute to the ecological fitness and pathogenicity of the producing strains. In this study, we employed a collection of 94 strains having various geographic, environmental, and clinical origins to study the distribution and evolution of phenazine genes in members of the genera Pseudomonas, Burkholderia, Pectobacterium, Brevibacterium, and Streptomyces. Our results confirmed the diversity of phenazine producers and revealed that most of them appear to be soil-dwelling and/or plant-associated species. Genome analyses and comparisons of phylogenies inferred from sequences of the key phenazine biosynthesis (phzF) and housekeeping (rrs, recA, rpoB, atpD, and gyrB) genes revealed that the evolution and dispersal of phenazine genes are driven by mechanisms ranging from conservation in Pseudomonas spp. to horizontal gene transfer in Burkholderia spp. and Pectobacterium spp. DNA extracted from cereal crop rhizospheres and screened for the presence of phzF contained sequences consistent with the presence of a diverse population of phenazine producers in commercial farm fields located in central Washington state, which provided the first evidence of United States soils enriched in indigenous phenazine-producing bacteria. PMID- 20008173 TI - Direct link between toluene degradation in contaminated-site microcosms and a Polaromonas strain. AB - Stable isotope probing (SIP) was used to identify the aerobic toluene-degrading microorganism in soil microcosms. Several approaches (terminal restriction fragment length polymorphism, 16S rRNA gene sequencing, and quantitative PCR) provided evidence that the microorganism responsible was a member of the genus Polaromonas and could grow on toluene. This microorganism also transformed benzene, but not m-xylene or cis-dichloroethene. PMID- 20008174 TI - Lysogeny and sporulation in Bacillus isolates from the Gulf of Mexico. AB - Eleven Bacillus isolates from the surface and subsurface waters of the Gulf of Mexico were examined for their capacity to sporulate and harbor prophages. Occurrence of sporulation in each isolate was assessed through decoyinine induction, and putative lysogens were identified by prophage induction by mitomycin C treatment. No obvious correlation between ability to sporulate and prophage induction was found. Four strains that contained inducible virus-like particles (VLPs) were shown to sporulate. Four strains did not produce spores upon induction by decoyinine but contained inducible VLPs. Two of the strains did not produce virus-like particles or sporulate significantly upon induction. Isolate B14905 had a high level of virus-like particle production and a high occurrence of sporulation and was further examined by genomic sequencing in an attempt to shed light on the relationship between sporulation and lysogeny. In silico analysis of the B14905 genome revealed four prophage-like regions, one of which was independently sequenced from a mitomycin C-induced lysate. Based on PCR and transmission electron microscopy (TEM) analysis of an induced phage lysate, one is a noninducible phage remnant, one may be a defective phage-like bacteriocin, and two were inducible prophages. One of the inducible phages contained four putative transcriptional regulators, one of which was a SinR-like regulator that may be involved in the regulation of host sporulation. Isolates that both possess the capacity to sporulate and contain temperate phage may be well adapted for survival in the oligotrophic ocean. PMID- 20008175 TI - Bacteria associated with immunoregulatory cells in mice. AB - This study examined bacteria-immune interactions in a mouse model possessing microbiota-dependent immune regulatory features similar to those occurring in human atopy, colitis, and immune regulation. Associations between the abundance of several bacterial phylotypes and immunoregulatory target cell types were identified, suggesting that they may play a role in these phenotypes. PMID- 20008177 TI - Introduction to surrogates and evidence-based mini-reviews. PMID- 20008176 TI - Darwin and evolutionary tales in leukemia. The Ham-Wasserman Lecture. AB - All cancers evolve by a process of genetic diversification and "natural selection" akin to the process first described by Charles Darwin for species evolution. The evolutionary, natural history of childhood acute lymphoblastic leukemia (ALL) is almost entirely covert, clinically silent and well advanced by the point of diagnosis. It has, however, been possible to backtrack this process by molecular scrutiny of appropriate clinical samples: (i) leukemic clones in monozygotic twins that are either concordant or discordant for ALL; (ii) archived neonatal blood spots or Guthrie cards from individuals who later developed leukemia; and (iii) stored, viable cord blood cells. These studies indicate prenatal initiation of leukemia by chromosome translocation and gene fusion (or hyperdiploidy) and the post-natal acquisition of multiple, gene copy number alterations (CNAs), mostly deletions. The prenatal or first "hit" occurs very commonly, exceeding the clinical rate of ALL by some 100x and indicating a low rate of penetrance or evolutionary progression. The acquisition of the critical, secondary CNAs requires some Darwinian selective advantage to expand numbers of cells at risk, and the cytokine TGF beta is able to exercise this function. The clonal architecture of ALL has been investigated by single cell analysis with multicolor probes to mutant genes. The data reveal not a linear sequence of mutation acquisition with clonal succession but rather considerable complexity with a tree-like or branching structure of genetically distinct subclones very reminiscent of Darwin's original 1837 evolutionary divergence diagram. This evolutionary pattern has important implications for stem cells in ALL, for the origins of relapse and for therapeutic targeting. PMID- 20008178 TI - Newborn screening for non-sickling hemoglobinopathies. AB - The hemoglobinopathies encompass a heterogeneous group of disorders associated with mutations in both the alpha-globin and beta-globin genes. Non-sickling disorders are found primarily in individuals of Mediterranean, Asian and Southeast Asian ancestry. With rapid growth in the Asian and Hispanic segments of the US population, the geographic distribution of hemoglobinopathies is expected to become significantly different from what it is today. The epidemiologic changes in the prevalence of non-sickling hemoglobin disorders have important implications for future public health programs, including newborn screening. The purpose of newborn screening for hemoglobinopathies is to identify clinically significant disorders and provide early education and specialized care prior to the onset of clinical symptoms. Although newborn screening for sickle cell disease is mandated in all states, screening for non-sickling hemoglobinopathies is directed in only one state and limited to reporting of a presumptive diagnosis in most other states. Early delivery of comprehensive care, as well as new and potentially curative therapies, has significantly improved the prognosis for affected patients. This review will consider the increasing prevalence of once uncommon hemoglobinopathies in the US, highlighting the rationale for expanding newborn screening beyond sickle cell disorders. PMID- 20008179 TI - Hb H disease: clinical course and disease modifiers. AB - Hemoglobin H (Hb H) disease is the most common form of thalassemia intermedia and has many features that require careful consideration in management. In the majority of cases, Hb H disease results from double heterozygosity for alpha(0) thalassemia due to deletions that remove both linked alpha-globin genes on chromosome 16, and deletional alpha(+)-thalassemia from single alpha-globin gene deletions (--/-alpha). However, Hb H disease may occur from interactions between alpha(0)-thalassemia with non-deletional mutations (alpha(T)alpha or alpha(T)) or with abnormal hemoglobins such as Hb Constant Spring, Hb Pakse, Hb Quong Sze, and Hb Pak Num Po. In a steady state, patients with Hb H diseases have hemoglobin levels around 9 to 10 g/dL; however, during hemolytic crisis, which frequently develops in or after acute infections with high fever, the hemoglobin level may drop significantly and patients can develop shock or renal shutdown. Even though splenectomy leads to significant elevation of hemoglobin levels, it is not recommended because the majority of patients do well with said steady-state hemoglobin levels. Patients with non-deletional Hb H disease are usually more anemic with significant splenomegaly, and some may require regular blood transfusions and be even as severe as "Hb H hydrops fetalis." However, there is no clear genotype-phenotype correlation associated with this severe clinical syndrome since patients with identical genotypes do not necessary show the same severity. This suggests that other genetic and environmental factors play a role in modifying the degree of clinical severity in patients with non-deletional Hb H disease. PMID- 20008180 TI - Alpha thalassemia major--new mutations, intrauterine management, and outcomes. AB - Alpha thalassemia disorders are a group of hereditary anemias caused by absent or decreased production of the alpha chain of hemoglobin. Hemoglobin Bart's hydrops fetalis is usually a fatal in-utero disease caused by absence of the alpha genes. However, the molecular and genotypic expression of hemoglobin Bart's varies and increasing numbers of births are being reported. Population screening and prenatal diagnosis of at-risk couples is essential but often not performed. Most affected pregnancies are often undetected, resulting in severe fetal and maternal complications. Noninvasive monitoring by Doppler ultrasonagraphy with intrauterine transfusion therapy has changed the prognosis for this disorder. These advances in intrauterine and postnatal therapy have resulted in ethical dilemmas for the family and the provider. PMID- 20008181 TI - Asthma and sickle cell disease: two distinct diseases or part of the same process? AB - A physician diagnosis of asthma in children and adults with sickle cell disease (SCD) has been associated with increased rates of pain and acute chest syndrome (ACS) episodes and premature death. Despite the clinical significance of a doctor's diagnosis of asthma in individuals with SCD, the criteria for a physician diagnosis of asthma are not well defined. Many features of asthma are common in individuals with SCD, including symptoms of wheezing, obstructive lung disease and airway hyper-responsiveness. However, it is not clear if these signs and symptoms of asthma reflect a physician diagnosis of asthma, or if these asthma features are related to SCD. Further complicating the diagnosis of asthma in children with SCD is the significant overlap in clinical manifestations between an asthma exacerbation and an ACS episode. Evidence supporting the concept that asthma and SCD are separate co-morbid conditions includes a similar prevalence of asthma between children with SCD and those in the general population and the observation that asthma is inherited in a familial pattern in the families of children with SCD. In contrast, there is significant evidence that asthma-like features may be associated with SCD without a diagnosis of asthma, including a higher than expected prevalence of airway hyper responsiveness and obstructive lung disease. Regardless of whether SCD and asthma are distinct or overlapping co-morbid conditions, we recommend a systematic and complete evaluation of asthma when the diagnosis is suspected or when patients have multiple episodes of pain or ACS. PMID- 20008182 TI - Novel therapies in sickle cell disease. AB - Despite an increased understanding of the pathophysiology of sickle cell disease (SCD), there remains a paucity of available agents for the prevention and treatment of specific SCD-related complications. Recently, there has been significant progress in the development of novel drugs for this disease. These agents, which increase the production of fetal hemoglobin, improve red blood cell hydration, increase the availability of nitric oxide and possess anti inflammatory effects, are in varying stages of clinical development. With the complex pathophysiology of SCD, it is unlikely that a single agent will prevent or treat all the sequelae of this disease. As a result, patients may benefit from treatment with a combination of agents that possess different mechanisms of action. This overview discusses selected novel agents that appear promising in SCD. PMID- 20008183 TI - Advances in the use of hydroxyurea. AB - Clinical experience with hydroxyurea for patients with sickle cell disease (SCD) has been accumulating for the past 25 years. The bulk of the current evidence suggests that hydroxyurea is well-tolerated, safe, and efficacious for most patients with SCD. Hydroxyurea has proven clinical efficacy for reducing acute vaso-occlusive events including pain episodes and acute chest syndrome. Salutary effects on hematological parameters include increases in fetal hemoglobin (HbF), hemoglobin, and MCV; also significant decreases occur in WBC, ANC, reticulocytes, LDH, and bilirubin. Treatment with hydroxyurea is usually considered for patients with recurrent vaso-occlusive events, but additional indications for treatment may include laboratory markers of disease severity and evidence of chronic organ dysfunction. Ten years ago, the US Food and Drug Administration approved hydroxyurea for adult patients with clinically severe SCD; however, its use in children remains off-label. Despite the large body of evidence regarding its efficacy and safety, hydroxyurea is currently prescribed only sparingly for patients with SCD and therefore has only limited effectiveness for this disorder; barriers to its use need to be identified and overcome. PMID- 20008184 TI - Drug-induced immune hemolytic anemia. AB - Drug-induced immune hemolytic anemia (DIIHA) is rare, and a specialized laboratory is often required to provide the optimal serological tests to confirm the diagnosis. The most common drugs associated with DIIHA and the hypotheses for the mechanisms thought to be involved have changed during the last few decades. The drugs most frequently associated with DIIHA at this time are cefotetan, ceftriaxone, and piperacillin. DIIHA is attributed most commonly to drug dependent antibodies that can only be detected in the presence of drug (eg, cephalosporin antibodies). DIIHA can also be associated with drug-independent antibodies; such antibodies do not need drug to be present to obtain in vitro reactions (eg, fludarabine). In these latter cases, the drug affects the immune system, causing production of red cell (RBC) autoantibodies; the clinical and laboratory findings are identical to autoimmune hemolytic anemia (AIHA), other than the remission associated with discontinuing the drug. Some of the mechanisms involved in DIIHA are controversial. The most acceptable one involves drugs, like penicillin, that covalently bind to proteins (eg, RBC membrane proteins); RBCs become coated with drug in vivo and, a drug antibody (usually IgG) attaches to the drug-coated RBCs that are subsequently cleared by macrophages. The most controversial is the so-called immune complex mechanism, which has been revised to suggest that most drugs are capable of binding to RBC membrane proteins, but not covalently like penicillins. The combined membrane plus drug can create an immunogen; the antibodies formed can be IgM or IgG and often activate complement, leading to acute intravascular lysis and sometimes renal failure; fatalities are more common in this group. It is still unknown why and how some drugs induce RBC autoantibodies, sometimes causing AIHA. PMID- 20008185 TI - Immune hemolytic anemia--selected topics. AB - Autoimmune hemolytic anemia (AIHA) is most often idiopathic. However, in recent years, AIHA has been noted with increased incidence in patients receiving purine nucleoside analogues for hematologic malignancies; it has also been described as a complication of blood transfusion in patients who have also had alloimmunization. As the technology of hematopoietic stem cell transplantation has become more widespread, immune hemolysis in the recipients of ABO-mismatched products has become better recognized. The syndrome is caused by passenger lymphocytes transferred from the donor and, although transient, can be quite severe. A similar syndrome has been observed in recipients of solid organ transplants when there is ABO-incompatibility between donor and recipient. Venous thromboembolism is a little-recognized, though likely common, complication of AIHA, and may in some instances be related to coexistent antiphospholipid antibodies. While AIHA is a well-documented complication of malignant lymphoproliferative disorders, lymphoproliferative disorders may also paradoxically appear as a consequence of AIHA. A number of newer options are available for treatment of AIHA in patients refractory to corticosteroids and splenectomy. Newer immunosuppressives such as mycophenolate mofetil may have a role in such cases. Considerable experience has been accumulating in the last few years with monoclonal antibody therapy, mainly rituximab, in difficult AIHA cases; it appears to be a safe and effective option. PMID- 20008187 TI - Factor XI deficiency--resolving the enigma? AB - The management of factor XI deficiency is not straightforward for three reasons: firstly, the role of this factor in the coagulation pathway is not clearly understood; secondly, the bleeding tendency, although mild, is unpredictable and does not clearly relate to the factor XI level; and thirdly, all treatment products, although available, have some potentially serious side effects. These factors (or enigmas) contribute to the variable management of patients with this coagulation factor deficiency, but recent research is helping to clarify some of these areas. PMID- 20008186 TI - Malaria, erythrocytic infection, and anemia. AB - Malaria is a major world health problem. It results from infection of parasites belonging to the genus Plasmodium. Plasmodium falciparum and Plasmodium vivax cause the major human malarias, with P falciparum being the more virulent. During their blood stages of infection, both P falciparum and P vivax induce anemia. Severe malarial anemia caused by P falciparum is responsible for approximately a third of the deaths associated with disease. Malarial anemia appears to be multi factorial. It involves increased removal of circulating erythrocytes as well as decreased production of erythrocytes in the bone marrow. The molecular mechanisms underlying malarial anemia are largely unknown. Over the last five years, malaria parasite ligands have been investigated for their remodeling of erythrocytes and possible roles in destruction of mature erythrocytes. Polymorphisms in cytokines have been associated with susceptibility to severe malarial anemia: these cytokines and malaria "toxins" likely function by perturbing erythropoiesis. Finally a number of co-infections increase susceptibility to malarial anemia, likely because they exacerbate inflammation caused by malaria. Because of the complexities involved, the study of severe malarial anemia may need a "systems approach" to yield comprehensive understanding of defects in both erythropoiesis and immunity associated with disease. New and emerging tools such as (i) mathematical modeling of the dynamics of host control of malarial infection, (ii) ex vivo perfusion of human spleen to measure both infected and uninfected erythrocyte retention, and (iii) in vitro development of erythroid progenitors to dissect responsiveness to cytokine imbalance or malaria toxins, may be especially useful to develop integrated mechanistic insights and therapies to control this major and fatal disease pathology. PMID- 20008188 TI - Low von Willebrand factor: sometimes a risk factor and sometimes a disease. AB - A sufficiently low level of von Willebrand factor (VWF) predisposes to bleeding that can be quite serious, and low VWF is a diagnostic feature of von Willebrand disease (VWD) type 1, which is characterized by partial quantitative deficiency of VWF. Recent groundbreaking studies of patients with VWD type 1 have delineated several pathophysiologic mechanisms that determine the plasma concentration of VWF, but the relationship between VWF level and the likelihood of bleeding remains less well understood. In part, this problem reflects the broad range of VWF levels in the population, so that the distinction between "normal" and "low" is arbitrary. The risk of bleeding certainly increases as the VWF level decreases, but the relationship is not very strong until the VWF level is very low. Furthermore, mild bleeding symptoms are common in apparently healthy populations and have many causes other than defects in VWF, which can make it impossible to attribute bleeding to any single factor, such as low VWF. These difficulties might be resolved by an epidemiologic approach to VWF and other risk factors for bleeding, analogous to how physicians manage multiple risk factors for cardiovascular disease or venous thromboembolism. PMID- 20008189 TI - Optimizing treatment of von Willebrand disease by using phenotypic and molecular data. AB - The wide clinical spectrum of von Willebrand disease (VWD), its complex pathophysiology and its classification into distinct quantitative (type 1 or type 3) and qualitative (type 2) types with further subtle distinctions have prevented most clinicians from establishing a straightforward approach to diagnosing and treating this inherited bleeding disorder. The results of studies involving large cohorts of patients with a wide range of bleeding manifestations and variable von Willebrand factor (VWF) reduction have recently become available. These data have allowed the proposal of minimal criteria for a clinically useful diagnosis and for differentiating patients with mild VWD from subjects with borderline or only slightly reduced VWF levels who will not benefit from a specific diagnosis. These criteria are based on measurement of VWF ristocetin cofactor (VWF:RCo), VWF antigen (VWF:Ag), factor VIII and a standardized bleeding score (BS). Demonstration of the inheritance of the disorder could help to classify patients for whom insufficient hemostatic challenges may produce a falsely reassuring BS (like in children). Using this approach, mild VWD appears to be mostly composed of type 1 cases. Complemented by the results of desmopressin trial infusion, these parameters form the basis for a clinically oriented classification of all forms of VWD and may be useful for selecting the best treatment according to the severity of the disease. Although few molecular data have revealed practical utility, there is no doubt that the clarification of the molecular pathophysiology of VWD has allowed the unification of this complex disorder into a simple conceptual framework. This framework underlies the proposed utilization of simple phenotypic markers for optimizing treatments in individual patients. PMID- 20008190 TI - Hemophagocytic lymphohistiocytosis (HLH) and related disorders. AB - Hemophagocytic lymphohistiocytosis (HLH), which has many genetic causes, is characterized by multi-system inflammation. HLH is a reactive process resulting from prolonged and excessive activation of antigen presenting cells (macrophages, histiocytes) and CD8(+) T cells. Hemophagocytosis, which is mediated through the CD163 heme-scavenging receptor, is a hallmark of activated macrophages/histiocytes and is the characteristic finding for which the disorder was named. The majority of genetic causes identified to date affect the cytotoxic function of NK and T cells, crippling immunologic mechanisms that mediate natural immune contraction. The predominant clinical findings of HLH are fevers (often hectic and persistent), cytopenias, hepatitis and splenomegaly. Due to the life threatening implications of the diagnosis of genetically determined HLH, antiinflammatory therapy, often consisting of steroids, etoposide or antithymocyte globulin (ATG), should be instituted promptly, followed by curative hematopoietic cell transplantation. Secondary HLH, associated with autoimmune disorders or viral infections in teens and adults, also carries a significant mortality rate and should be managed in consultation with specialists familiar with the diagnosis and treatment of such disorders. PMID- 20008191 TI - New insights into the biology of Wiskott-Aldrich syndrome (WAS). AB - The Wiskott-Aldrich syndrome (WAS) is an X-linked immunodeficiency disease with a characteristic clinical phenotype that includes thrombocytopenia with small platelets, eczema, recurrent infections due to immunodeficiency, and an increased incidence of autoimmune manifestations and malignancies. The identification of the molecular defect in the WAS gene has broadened the clinical spectrum of disease to include chronic or intermittent X-linked thrombocytopenia (XLT), a relatively mild form of WAS, and X-linked neutropenia (XLN) due to an arrest of myelopoiesis. The pathophysiological mechanisms relate to defective actin polymerization in hematopoietic cells as a result of deficient or dysregulated activity of the WAS protein (WASp). The severity of disease is variable and somewhat predictable from genotype. Treatment strategies therefore range from conservative through to early definitive intervention by using allogeneic hematopoietic stem cell transplantation and potentially somatic gene therapy. All aspects of the condition from clinical presentation to molecular pathology and basic cellular mechanisms have been reviewed recently. PMID- 20008192 TI - Primary immunodeficiencies (PIDs) presenting with cytopenias. AB - Autoimmune manifestations are increasingly being recognized as a component of several forms of primary immunodeficiencies (PID). Defects in purging of self reactive T and B cells, impaired Fas-mediated apoptosis, abnormalities in development and/or function of regulatory T cells, and persistence of immune activation as a result of inability to clear infections have been shown to account for this association. Among autoimmune manifestations in patients with PID, cytopenias are particularly common. Up to 80% of patients with autoimmune lymphoproliferative syndrome (ALPS) have autoantibodies, and autoimmune hemolytic anemia and immune thrombocytopenia have been reported in 23% and 51% of ALPS patients, and may even mark the onset of the disease. ALPS-associated cytopenias are often refractory to conventional treatment and represent a therapeutic challenge. Autoimmune manifestations occur in 22% to 48% of patients with common variable immunodeficiencies (CVIDs), and are more frequent among CVID patients with splenomegaly and granulomatous disease. Finally, autoimmune cytopenias have been reported also in patients with combined immunodeficiency. In particular, autoimmune hemolytic anemia is very common among infants with nucleoside phosphorylase deficiency. While immune suppression may be beneficial in these cases, full resolution of the autoimmune manifestations ultimately depends on immune reconstitution, which is typically provided by hematopoietic cell transplantation. PMID- 20008193 TI - Determinants of platelet number and regulation of thrombopoiesis. AB - Our understanding of thrombopoiesis has improved greatly in the last two decades with the availability of in vitro assays of megakaryocyte progenitor cell growth, with the cloning and characterization of stem cell factor (SCF) and thrombopoietin (Tpo), the latter the primary humoral regulator of this process, and with the generation of genetically altered murine models of thrombopoietic failure and excess. While SCF affects developmentally early aspects of megakaryocyte growth, Tpo affects nearly all aspects of platelet production, from hematopoietic stem cell (HSC) self-renewal and expansion, through stimulation of megakaryocyte progenitor cell proliferation, to supporting their maturation into platelet-producing cells. The molecular and cellular mechanisms through which the marrow microenvironment and humoral mediators affect platelet production provide new insights into the interplay between intrinsic and extrinsic influences on hematopoiesis, and highlight new opportunities to translate basic biology into clinical advances. PMID- 20008194 TI - Drug-induced thrombocytopenia: pathogenesis, evaluation, and management. AB - Although drugs are a common cause of acute immune-mediated thrombocytopenia in adults, the drug etiology is often initially unrecognized. Most cases of drug induced thrombocytopenia (DITP) are caused by drug-dependent antibodies that are specific for the drug structure and bind tightly to platelets by their Fab regions but only in the presence of the drug. A comprehensive database of 1301 published reports describing 317 drugs, available at www.ouhsc.edu/platelets, provides information on the level of evidence for a causal relation to thrombocytopenia. Typically, DITP occurs 1 to 2 weeks after beginning a new drug or suddenly after a single dose when a drug has previously been taken intermittently. However, severe thrombocytopenia can occur immediately after the first administration of antithrombotic agents that block fibrinogen binding to platelet GP IIb-IIIa, such as abciximab, tirofiban, and eptifibatide. Recovery from DITP usually begins within 1 to 2 days of stopping the drug and is typically complete within a week. Drug-dependent antibodies can persist for many years; therefore, it is important that the drug etiology be confirmed and the drug be avoided thereafter. PMID- 20008195 TI - Thrombocytosis. AB - Major progress in understanding the pathogenesis in patients with thrombocytosis has been made by identifying mutations in the key regulators of thrombopoietin: the thrombopoietin receptor MPL and JAK2. Together, these mutations can be found in 50% to 60% of patients with essential thrombocythemia or primary myelofibrosis and in 10% to 20% of hereditary thrombocytosis. A decrease in expression of the Mpl protein can cause thrombocytosis even in the absence of mutations in the coding sequence, due to a shift in the balance between stimulation of signaling in megakaryopoiesis and removal of thrombopoietin by receptor mediated internalization in platelets. When present in a heterozygous state the JAK2-V617F mutation preferentially stimulates megakaryopoiesis and in most cases manifests as essential thrombocythemia (ET), whereas homozygous JAK2-V617F reduces megakaryopoiesis in favor of increased erythropoiesis, resulting in polycythemia vera and/or myelofibrosis. In 30% to 40% of patients with ET or primary myelofibrosis (PMF) and in 80% to 90% of pedigrees with hereditary thrombocytosis the disease-causing gene remains unknown. Ongoing genetic and genomic screens have identified genes that, when mutated, can cause thrombocytosis in mouse models. A more complete picture of the pathways that regulate megakaryopoisis and platelet production will be important for finding new ways of controlling platelet production in patients with thrombocytosis. PMID- 20008196 TI - Transfusion in the age of molecular diagnostics. AB - DNA-based tests are increasingly being used to predict a blood group phenotype to improve transfusion medicine. This is possible because genes encoding 29 of the 30 blood group systems have been cloned and sequenced, and the molecular bases associated with most antigens have been determined. RBCs carrying a particular antigen, if introduced into the circulation of an individual who lacks that antigen (through transfusion or pregnancy), can elicit an immune response. It is the antibody from such an immune response that causes problems in clinical practice and the reason why antigen-negative blood is required for safe transfusion. The classical method of testing for blood group antigens and antibodies is hemagglutination; however, it has certain limitations, some of which can be overcome by testing DNA. Such testing allows conservation of antibodies for confirmation by hemagglutination of predicted antigen-negativity. High-throughput platforms provide a means to test relatively large numbers of donors, thereby opening the door to change the way antigen-negative blood is provided to patients and to prevent immunization. This review summarizes how molecular approaches, in conjunction with conventional hemagglutination, can be applied in transfusion medicine. PMID- 20008197 TI - Molecular biology of the Rh system: clinical considerations for transfusion in sickle cell disease. AB - The last decade has witnessed an abundance of information detailing the genetic diversity of the RH locus which has exceeded all estimates predicted by serology. Well over 120 RHD and over 60 different RHCE alleles have been documented, and new alleles are still being discovered. For clinical transfusion medicine, RH genetic testing can now be used to determine RHD zygosity, resolve D antigen status, and detect altered RHD and RHCE genes in individuals at risk for producing antibodies to high-incidence Rh antigens, particularly patients with sickle cell disease (SCD). PMID- 20008198 TI - Mechanisms of anti-D action in the prevention of hemolytic disease of the fetus and newborn. AB - Anti-D is routinely and effectively used to prevent hemolytic disease of the fetus and newborn (HDFN) caused by the antibody response to the D antigen on fetal RBCs. Anti-D is a polyclonal IgG product purified from the plasma of D alloimmunized individuals. The mechanism of anti-D has not been fully elucidated. Antigenic epitopes are not fully masked by anti-D and are available for immune system recognition. However, a correlation has frequently been observed between anti-D-mediated RBC clearance and prevention of the antibody response, suggesting that anti-D may be able to destroy RBCs without triggering the adaptive immune response. Anti-D-opsonized RBCs may also elicit inhibitory FcgammaRIIB signaling in B cells and prevent B cell activation. The ability of antigen-specific IgG to inhibit antibody responses has also been observed in a variety of animal models immunized with a vast array of different antigens, such as sheep RBCs (SRBC). This effect has been referred to as antibody-mediated immune suppression (AMIS). In animal models, IgG inhibits the antibody response, but the T-cell response and memory may still be intact. IgG does not mask all epitopes, and IgG-mediated RBC clearance or FcgammaRIIB-mediated B-cell inhibition do not appear to mediate the AMIS effect. Instead, IgG appears to selectively disrupt B cell priming, although the exact mechanism remains obscure. While the applicability of animal models of AMIS to understanding the true mechanism of anti-D remains uncertain, the models have nevertheless provided us with insights into the possible IgG effects on the immune response. PMID- 20008199 TI - Hereditary hemochromatosis: insights from the Hemochromatosis and Iron Overload Screening (HEIRS) Study. AB - Hemochromatosis comprises a group of inherited disorders resulting from mutations of genes involved in regulating iron metabolism. The multicenter, multi-ethnic Hemochromatosis and Iron Overload Screening (HEIRS) Study screened approximately 100,000 participants in the US and Canada, testing for HFE mutations, serum ferritin and transferrin saturation. As in other studies, HFE C282Y homozygosity was common in Caucasians but rare in other ethnic groups, and there was a marked heterogeneity of disease expression in C282Y homozygotes. Nevertheless, this genotype was often associated with elevations of serum ferritin and transferrin saturation and with iron stores of more than four grams in men but not in women. If liver biopsy was performed, in some cases because of evidence of hepatic dysfunction, fibrosis or cirrhosis was often found. Combined elevations of serum ferritin and transferrin saturation were observed in non-C282Y homozygotes of all ethnic groups, most prominently Asians, but not often with iron stores of more than four grams. Future studies to discover modifier genes that affect phenotypic expression in C282Y hemochromatosis should help identify patients who are at greatest risk of developing iron overload and who may benefit from continued monitoring of iron status to detect progressive iron loading. PMID- 20008201 TI - Non-invasive assessment of tissue iron overload. AB - In recent years, there has been increasing interest in non-invasive iron measurement, especially of the liver and heart, in patients with iron overload. Serum ferritin still remains an essential monitoring parameter in intervals between liver iron measurements; however, confounding factors such as inflammation, chelation treatment changes and the specific disease have to be taken into account. Liver iron measurements can now routinely be performed in clinical applications either by quantitative magnetic resonance imaging (MRI) using the transverse magnetic relaxation rate R(2) or R(2)* (1/T(2)*) or by biomagnetic liver susceptometry. For iron measurements in the heart, the single breathhold multi-echo MRI-R(2)* method has become a standard modality and is now applied in clinical settings beyond research studies. In other tissues like the pancreas, pituitary, and brain, different MRI methods are employed, but their clinical benefit has yet to be proven. PMID- 20008202 TI - Heparin-induced thrombocytopenia: when a low platelet count is a mandate for anticoagulation. AB - Heparin-induced thrombocytopenia (HIT) is an immune-mediated disorder caused by the development of antibodies to platelet factor 4 (PF4) and heparin. The thrombocytopenia is typically moderate, with a median platelet count nadir of approximately 50 to 60 x 10(9) platelets/L. Severe thrombocytopenia has been described in patients with HIT, and in these patients antibody levels are high and severe clinical outcomes have been reported (eg, disseminated intravascular coagulation with microvascular thrombosis). The timing of the thrombocytopenia in relation to the initiation of heparin therapy is critically important, with the platelet count beginning to drop within 5 to 10 days of starting heparin. A more rapid drop in the platelet count can occur in patients who have been recently exposed to heparin (within the preceding 3 months), due to preformed anti heparin/PF4 antibodies. A delayed form of HIT has also been described that develops within days or weeks after the heparin has been discontinued. In contrast to other drug-induced thrombocytopenias, HIT is characterized by an increased risk for thromboembolic complications, primarily venous thromboembolism. Heparin and all heparin-containing products should be discontinued and an alternative, non-heparin anticoagulant initiated. Alternative agents that have been used effectively in patients with HIT include lepirudin, argatroban, bivalirudin, and danaparoid, although the last agent is not available in North America. Fondaparinux has been used in a small number of patients with HIT and generally appears to be safe. Warfarin therapy should not be initiated until the platelet count has recovered and the patient is systemically anticoagulated, and vitamin K should be administered to patients receiving warfarin at the time of diagnosis of HIT. PMID- 20008200 TI - Molecular mechanisms of normal iron homeostasis. AB - Humans possess elegant control mechanisms to maintain iron homeostasis by coordinately regulating iron absorption, iron recycling, and mobilization of stored iron. Dietary iron absorption is regulated locally by hypoxia inducible factor (HIF) signaling and iron-regulatory proteins (IRPs) in enterocytes and systematically by hepatic hepcidin, the central iron regulatory hormone. Hepcidin not only controls the rate of iron absorption but also determines iron mobilization from stores through negatively modulating the function of ferroportin, the only identified cellular iron exporter to date. The regulation of hepatic hepcidin is accomplished by the coordinated activity of multiple proteins through different signaling pathways. Recent studies have greatly expanded the knowledge in the understanding of hepcidin expression and regulation by the bone morphogenetic protein (BMP) signaling, the erythroid factors, and inflammation. In this review, we mainly focus on the roles of recently identified proteins in the regulation of iron homeostasis. PMID- 20008203 TI - Antiphospholipid antibody syndrome. AB - The antiphospholipid antibody syndrome (APS) is defined by the persistent presence of antiphospholipid antibodies in patients with recurrent venous or arterial thromboembolism or pregnancy morbidity. Anti-thrombotic therapy is the mainstay of treatment given the high risk of recurrent thromboembolism that characterizes this condition. Despite the prothrombotic nature of APS, thrombocytopenia is present in a proportion of patients. which can complicate management and limit the use of antithrombotic therapy. The mechanism of APS associated thrombocytopenia is multifactorial and its relation to thrombotic risk poorly characterized. However, the presence of thrombocytopenia does not appear to reduce thrombotic risk in patients with APS, who can develop thromboembolic complications necessitating antithrombotic treatment. In these cases, treatment of the thrombocytopenia may be necessary to facilitate administration of antithrombotic agents. Clinical trials have demonstrated that patients with antiphospholipid antibodies and venous thromboembolism should be treated with vitamin K antagonists (warfarin); that ischemic stroke may be treated with aspirin or warfarin; and that women with recurrent pregnancy loss should receive prophylactic-dose heparin and aspirin. However, application of these trial results to patients with APS-associated thrombocytopenia can be challenging since there are limited data on the optimal use of antithrombotic agents in this setting. Issues such as determining the platelet threshold at which antithrombotic agents can be safely used and managing patients with both bleeding and thromboembolic complications remain unresolved. Ultimately the risks and benefits of antithrombotic therapy, balanced against the severity of the thrombocytopenia and its potential bleeding risks, need to be assessed using an individualized patient approach. PMID- 20008204 TI - Thrombocytopenia and thrombosis in disseminated intravascular coagulation (DIC). AB - Disseminated intravascular coagulation (DIC) is the physiologic result of pathologic overstimulation of the coagulation system. Despite multiple triggers, a myriad of laboratory abnormalities, and a clinical presentation ranging from gross hemostatic failure to life-threatening thrombosis, or even both simultaneously, a simplified clinical approach augmented by a few readily available tests allows prompt identification of the process and elucidation of treatment opportunities. Platelet counts in DIC may be low, especially in acute sepsis-associated DIC, yet increased in malignancy-associated chronic DIC. Thrombotic risk is not a function of the platelet count, and thrombocytopenia does not protect the patient from thrombosis. The stratification of both thrombotic risk and hemorrhagic risk will be addressed. PMID- 20008205 TI - ASH evidence-based guidelines: should asymptomatic patients with antiphospholipid antibodies receive primary prophylaxis to prevent thrombosis? AB - A 35-year-old female presents with a prolonged activated partial thromboplastin time (aPTT) on routine testing, which is found to be due to a lupus anticoagulant. She has no medical issues, no personal or family history of thrombosis, no history of pregnancy loss, and no symptoms suggestive of an underlying rheumatologic disorder. She is a non-smoker and does not take oral contraceptives. You are asked to provide recommendations regarding the need for primary thromboprophylaxis. As you begin your literature search, you formulate the following clinical question: "In asymptomatic patients with antiphospholipid antibodies, does primary prophylaxis prevent thrombotic complications?" PMID- 20008206 TI - ASH evidence-based guidelines: is the IgG-specific anti-PF4/heparin ELISA superior to the polyspecific ELISA in the laboratory diagnosis of HIT? AB - You are asked to consult on a 76-year-old man admitted to the hospital with pneumonia and thrombocytopenia. Ten days before the current admission, he had undergone surgery to repair a small bowel obstruction. A preoperative platelet count had been normal. Following surgery, he received subcutaneous unfractionated heparin thromboprophylaxis until his discharge on post-operative day 5. In your differential diagnosis for the patient's thrombocytopenia, you consider heparin induced thrombocytopenia (HIT) and wish to order laboratory testing. In addition to a polyspecific anti-PF4/heparin ELISA for the diagnosis of HIT, your laboratory has recently begun to offer an IgG-specific ELISA. You wonder which of these assays performs better in the diagnosis of HIT. PMID- 20008207 TI - Pathogenesis of thrombosis. AB - The hemostatic process is a host defense mechanism to preserve the integrity of the closed high pressure circulatory system. This process must remain inactive but poised to minimize extravasation of blood from the vasculature following tissue injury. Given the complexity of the hemostatic mechanism, paradigms developed from biochemical and cell biological approaches have been revisited by studying thrombus formation in a live animal by intravital microscopy. Many of these paradigms have proven accurate, but others need to be reconsidered given the results of whole animal experiments. PMID- 20008208 TI - Risk of future arterial cardiovascular events in patients with idiopathic venous thromboembolism. AB - Venous and arterial thromboses have traditionally been considered distinct pathophysiologic entities. However, the two disorders have many features in common, and there is evidence that persons with venous thrombosis may be at greater risk for arterial events. The pathogenesis of both disorders includes endothelial injury, platelet activation, elevated levels of intrinsic clotting factors and inflammatory markers, increased fibrinogen, and impaired fibrinolysis. In addition, older age, obesity, dyslipidemia, and smoking predispose to both venous and arterial thrombosis. While the evidence that arterial disease is a risk factor for venous thrombosis is inconclusive, arterial disease does appear to occur with a modestly increased frequency in patients with a history of venous thromboembolism. Reported odds ratios in such patients were 1.2 for myocardial infarction, 1.3 for stroke, 2.3 for carotid plaque, and 4.3 for coronary calcification. Of note, in persons under age 40 with unprovoked venous thrombosis, the odds ratio for acute myocardial infarction was as high as 3.9. In general, however, venous disease is considered to be a weak risk factor for arterial thrombosis, and the use of agents specifically targeted to the prevention of heart attack or stroke in the majority of persons with VTE cannot be recommended at present. PMID- 20008209 TI - Antiplatelet agents. AB - The introduction of aspirin as an anti-thrombotic agent some 50 years ago has changed the therapeutic approach in cardiovascular medicine. Since platelets play a key role in the development of arterial thrombosis, antiplatelet drugs serve as a cornerstone in the prevention and the treatment of these conditions. After many years of a "monopoly" of aspirin, ADP receptor P2Y12 inhibitors were introduced with a significant improvement in clinical outcome. Nowadays dual antiplatelet therapy is the common practice for both acute events and secondary prevention in selected groups of patients. Another revolution was the development of potent inhibitors of the platelet integrin GPIIbIIIa, which significantly improved the outcome of percutaneous interventions (PCI), in cardiology. The improved efficacy of multiple-drug therapy is associated with an increased risk of bleeding, which raises the issue of the dosing of these drugs. Recently, numerous studies have reported a variable laboratory response to aspirin and clopidogrel, which correlates with clinical outcome. Several mechanisms have been proposed to cause this variable response, including genetic variability, disease burden and others. A major obstacle in this field is the lack of a standardized method for testing these responses, and thus some studies cannot be compared to others. Ongoing studies are currently investigating the potential translation of these observations into clinical practice. Such studies may lead to a change in the paradigm of antiplatelet therapy, where individual dose adjustment may improve efficacy and safety. Finally, a variety of new drugs are currently in different stages of development, including new P2Y12 receptor inhibitors, thromboxane receptor blockers, direct thrombin inhibitors and other signaling pathway inhibitors including oral GPIIbIIIa inhibitors. Thus, antiplatelet therapy is currently under intensive development toward multiple-drug therapy and personal dose adjustment, which may improve clinical outcome. PMID- 20008210 TI - ASH evidence-based guidelines: statins in the prevention of venous thromboembolism. AB - A 42-year-old woman under your care for the management of obesity calls you because she has a sister who had a pulmonary embolism. The patient recently read that statin medications are associated with a lower risk of venous thromboembolism (VTE) and wonders whether she should take a statin drug to prevent the development of pulmonary emboli. PMID- 20008211 TI - Pregnancy-associated thrombosis. AB - The main reason for the increased risk of thromboembolism in pregnancy is hypercoagulability, which has likely evolved to protect women from the bleeding challenges of miscarriage and childbirth. Women are at a 4- to 5-fold increased risk of thromboembolism during pregnancy and the postpartum period compared with when they are not pregnant. Eighty percent of the thromboembolic events in pregnancy are venous, with an incidence of 0.49 to 1.72 per 1000 pregnancies. Risk factors include a history of thrombosis, inherited and acquired thrombophilia, maternal age greater than 35, certain medical conditions, and various complications of pregnancy and childbirth. Despite the increased risk of venous thromboembolism (VTE) during pregnancy and the postpartum period, most women do not require anticoagulation. Candidates include women with current VTE, a history of VTE, thrombophilia and a history of poor pregnancy outcome, or risk factors for postpartum VTE. The intensity of the anticoagulation will depend on the indication and the monitoring will depend on the intensity. At the time of delivery, anticoagulation should be manipulated to reduce the risk of bleeding complications while minimizing the risk of thrombosis. There are no large trials of anticoagulants in pregnancy, and recommendations are based on case series, extrapolations from nonpregnant patients and the opinion of experts. Nonetheless, anticoagulants are believed to improve the outcome of pregnancy for women who have, or have had, VTE. PMID- 20008212 TI - Prevention of venous thromboembolism: consensus, controversies, and challenges. AB - The last 50 years have witnessed a multitude of publications evaluating the efficacy, safety and cost effectiveness of many different thromboprophylaxis interventions. There is widespread consensus that thromboprophylaxis safely reduces morbidity and mortality. More than 25 evidence-based guidelines, published since 1986, also recommend routine thromboprophylaxis in the majority of hospitalized patients. As a result, thromboprophylaxis is recognized as a key safety priority for hospitals. Some of the remaining areas of controversy that will be discussed in this paper include the role of individual risk assessments to determine thrombosis risk and prophylaxis, replacement of low-dose heparin by low-molecular-weight heparin (LMWH), the optimal duration of prophylaxis, the role of combined thromboprophylaxis modalities, the safety of anticoagulant prophylaxis with regional analgesia, the use of LMWHs in chronic renal insufficiency, and the emerging role of new oral anticoagulants as thromboprophylactic agents. Despite the overwhelming evidence supporting thromboprophylaxis, rates of thromboprophylaxis use remain far from optimal. Successful implementation strategies to bridge this knowledge:care gap are the most important current challenges in this area. These strategies must be multifaceted, utilizing local, systems-based approaches as well as legislation and incentives that reinforce best practices. PMID- 20008213 TI - Evaluation and management of stroke. AB - In contrast to myocardial infarction, stroke is a heterogeneous disease with multiple different causes. Determination of stroke mechanism is critical to choosing optimal therapy to prevent recurrence. This article reviews the diagnostic approach to stroke and prognostic and treatment implications of specific stroke mechanisms. Data on the role of antiplatelet and anticoagulant therapy in secondary stroke prevention, as well as the role of these agents and thrombolytic therapy in acute stroke treatment are reviewed. Situations of particular relevance to the practicing hematologist-stroke in the young, patients with multiple recurrent strokes, patients with abnormal hypercoagulable laboratory testing, and treatment of intracerebral hemorrhage following thrombolytic therapy-are discussed. PMID- 20008214 TI - ASH evidence-based guidelines: what is the role of inferior vena cava filters in the perioperative prevention of venous thromboembolism in bariatric surgery patients? AB - A 38-year-old morbidly obese male (BMI > 50 kg/m(2)) presents for an elective gastric bypass surgery. He has no personal or family history of venous thromboembolism or hypercoaguability. You are asked by his primary team whether he should receive a retrievable inferior vena cava filter preoperatively for venous thromboembolism prophylaxis. PMID- 20008215 TI - Integrative therapies for children with hematological malignancies. AB - Evidence for integrative therapies (IT) in children with hematological malignancies is slowly evolving. The ideal model of integrative pediatric oncology would offer IT modalities that are deemed safe and effective in conjunction with effective conventional medical treatments. Because of potential interactions, herbs and other dietary supplements should be used with caution, especially during active therapy. Health and wellness should be the emphasis, with IT therapies supporting health promotion and key disease prevention strategies for childhood cancer survivors. All uses, responses, and effects of IT therapies should be carefully documented. A desire to use IT therapies may be an effort to become an active participant in the healing process. Health care providers should encourage, not discourage, this partnership. PMID- 20008216 TI - Supplements for immune enhancement in hematologic malignancies. AB - This brief review aims to discuss the various cellular immunological aspects and related mechanisms of the use of specific components from traditional herbal medicines. We begin with lessons learned from thalidomide as an effective single drug with multiple mechanisms of action to treat multiple myeloma. Examples of "supplements" or integrative therapy will be drawn from arsenic trioxide, medicinal mushrooms including Coriolus vesicular and Ganoderma lucidum, followed by the discussion of beta-glucans affecting various immunological important cellular subsets. Different classes of compounds may enhance distinct immune cell populations that might contribute to a multi-targeted holistic effects on anti cancer treatment. Finally, we conclude by highlighting an herbal formulation PHY906 as a potential adjunct to chemotherapy that might become one of the first US Food and Drug Administration (FDA) approved oral herbal medicines for anti cancer adjunct treatment. PMID- 20008217 TI - The bridge between patient and doctor: the shift from CAM to integrative medicine. AB - Integrative medicine (IM) has become a major challenge for doctors and nurses, as well as psychologists and many other disciplines involved in the endeavor to help patients to better tolerate the burden of toxic therapies and give patients tools so they can actively participate in their "salutogenesis." IM encompasses psycho oncology, acupuncture, and physical and mental exercises to restore vital capacities lost due to toxic therapies; furthermore, it aims to replenish nutritional and metabolic deficits during and after cancer treatment. IM gains an ever increasing importance in the face of the rapidly growing number of cancer survivors demanding more than just evidence-based diagnostic and therapeutic strategies. IM has to prove its value and justification by filling the gap between unproven methods of alternative medicine, still used by many cancer patients, and academic conventional medicine, which often does not satisfy the emotional and spiritual needs of cancer patients. PMID- 20008218 TI - Clinical approach to marrow failure. AB - The treatment and medical management of aplastic anemia fundamentally differ between patients with inherited versus acquired marrow failure; however, the diagnosis of an inherited bone marrow failure syndrome is frequently obscure. Recent exciting advances in our understanding of the molecular pathophysiology of the inherited bone marrow failure syndromes have resulted in a profusion of new tests to aid in diagnosis. This in turn has raised questions regarding the appropriate choice of testing for the patient presenting with aplastic anemia. Important clues to the diagnosis of an inherited marrow failure syndrome may be gleaned from careful attention to the clinical history, physical exam, and laboratory workup. PMID- 20008219 TI - Telomeres and marrow failure. AB - Telomeres, repeat sequences at the ends of chromosomes, are protective chromosomal structures highly conserved from primitive organisms to humans. Telomeres inevitably shorten with every cell cycle, and telomere attrition has been hypothesized to be fundamental to normal senescence of cells, tissues, and organisms. Molecular mechanisms have evolved to maintain their length and protective function; telomerase (TERT) is a reverse transcriptase enzyme that uses an RNA molecule (TERC) as the template to elongate the 3' ends of telomeres. Shelterin is a collection of DNA-binding proteins that cover and protect telomeres. The recent discovery of inherited mutations in genes that function to repair telomeres as etiologic in a range of human diseases, which have clinical manifestations in diverse tissues, including the hematopoietic tissue, suggests that defects in telomere repair and protection can cause organ failure. Dyskeratosis congenita is the prototype of telomere diseases; it is characterized by bone marrow failure, mucocutaneous abnormalities, pulmonary fibrosis, liver cirrhosis, and increased susceptibility to cancer, including acute myeloid leukemia. Aplastic anemia, acute myeloid leukemia, and idiopathic pulmonary fibrosis also are associated with inherited mutations in telomere repair or protection genes. Additionally, telomere defects associate with predisposition to hematologic malignancy and epithelial tumors. Telomere erosion is abnormally rapid in patients with mutations in telomerase genes but also after hematopoietic stem cell transplant, and telomeres are naturally shorter in older individuals all conditions associated with higher rates of malignant diseases. In human tissue culture, short telomeres produce end-to-end chromosome fusion, nonreciprocal translocations, and aneuploidy. PMID- 20008220 TI - Congenital neutropenia. AB - Congenital neutropenia comprises a variety of genetically heterogeneous phenotypic traits. Molecular elucidation of the underlying genetic defects has yielded important insights into the physiology of neutrophil differentiation and function. Non-syndromic variants of congenital neutropenia are caused by mutations in ELA2, HAX1, GFI1, or WAS. Syndromic variants of congenital neutropenia may be due to mutations in genes controlling glucose metabolism (SLC37A4, G6PC3) or lysosomal function (LYST, RAB27A, ROBLD3/p14, AP3B1, VPS13B). Furthermore, defects in genes encoding ribosomal proteins (SBDS, RMRP) and mitochondrial proteins (AK2, TAZ) are associated with congenital neutropenia syndromes. Despite remarkable progress in the field, many patients with congenital neutropenia cannot yet definitively be classified by genetic terms. This review addresses diagnostic and therapeutic aspects of congenital neutropenia and covers recent molecular and pathophysiological insights of selected congenital neutropenia syndromes. PMID- 20008222 TI - Evolving paradigms in the therapy of Philadelphia-chromosome-negative acute lymphoblastic leukemia in adults. AB - Important studies challenging previous approaches to the treatment of adults with Philadelphia chromosome-negative acute lymphoblastic leukemia (ALL) have emerged in the past decade. Donor versus no donor comparisons of allogeneic transplant highlight a potent graft-versus-leukemia effect in ALL, and the application of reduced-intensity conditioning transplants may exploit this effect while reducing non-relapse mortality. The adoption of the use of pediatric intensity-type regimens in adolescents and young adults shows promise to improve outcomes in this population. New therapeutic targets such as mutations in NOTCH1 in T-cell ALL and CD22 in pre-B ALL are being exploited in clinical trials. The application of molecular techniques and flow cytometry to quantitate minimal residual disease will allow further stratification of patients by risk. Although the outcomes of adults with ALL lag behind the stunningly successful results seen in children, new paradigms and new discoveries bring hope that this disparity will steadily lessen. PMID- 20008221 TI - The role of NOTCH1 signaling in T-ALL. AB - The identification of activating mutations in NOTCH1 in over 50% of T-cell acute lymphoblastic leukemias (T-ALL) has generated major interest in the elucidation of the mechanisms of transformation downstream of oncogenic NOTCH and in the targeting of the NOTCH signaling pathway in this disease. Small molecule gamma secretase inhibitors (GSIs) block NOTCH1 signaling in T-ALL lymphoblasts, yet the clinical development of GSIs has been held back by the development of gastrointestinal toxicity and their weak antileukemic effects against human T ALL. However, new therapeutic strategies aiming to optimize the use of anti NOTCH1 therapies for T-ALL, including combination therapies with molecularly targeted drugs and glucocorticoids, have started to emerge as a result of improved understanding of the molecular mechanisms that mediate the effects of GSIs in leukemic cells and the intestinal epithelium. This review focuses on the molecular basis of NOTCH1-induced transformation, the mechanisms of action of oncogenic NOTCH1 and clinical significance of NOTCH1 mutations in T-ALL. PMID- 20008223 TI - Management of Philadelphia chromosome-positive acute lymphoblastic leukemia (Ph+ ALL). AB - The tyrosine kinase inhibitor (TKI) imatinib has become an integral part of front line therapy for Ph(+) ALL, with remission rates exceeding 90% irrespective of whether imatinib is given alone or combined with chemotherapy. Treatment outcome with imatinib-based regimens has improved compared with historic controls, but most patients who do not undergo allogeneic stem cell transplantation (SCT) eventually relapse. Acquired resistance on TKI treatment is associated with mutations in the bcr-abl tyrosine kinase domain in the majority of patients, and may be detected at low frequency prior to TKI treatment in a subset of patients. Second generation TKIs, eg, dasatinib and nilotinib, show activity against most of the bcr-abl tyrosine kinase domain (TKD) mutations involved in acquired imatinib resistance, but clinical benefit is generally short-lived. Accordingly, SCT in first complete remission (CR) is considered to be the best curative option. Molecular monitoring of minimal residual disease levels appears to have prognostic relevance and should be used to guide treatment. International standardization and quality control efforts are ongoing to ensure comparability of results. Mutation analysis during treatment relies increasingly on highly sensitive PCR techniques or denaturing HPLC and may assist in treatment decisions, eg, in case of molecular relapse. Results from current studies of second-generation TKI as front-line treatment for Ph(+) ALL are promising and show high molecular response rates, but follow-up is still too short to determine their impact on remission duration and long-term survival. Strategies to improve outcome after SCT include the pre-emptive use of imatinib, which appears to reduce the relapse rate. In patients ineligible for transplantation, novel concepts for maintenance therapy are needed. These could involve novel immunotherapeutic interventions and combinations of TKI. PMID- 20008224 TI - Independent prognostic factors for AML outcome. AB - Over the last three decades there have been dramatic advances in deciphering the cytogenetic and molecular lesions underlying the pathogenesis of acute myeloid leukemia (AML). These have not only afforded greater insights into disease biology, but also provided useful information predicting the likelihood of any given patient achieving and maintaining remission following conventional chemotherapy, leading to the development of risk-stratified treatment approaches. However, it is becoming increasingly apparent that AML is highly heterogeneous at the molecular level. Defining the individual genetic abnormalities or combinations of markers that provide significant independent prognostic information and establishing their respective relationships to other pre treatment characteristics that impact on outcome, such as age and presenting white blood cell count, presents a major ongoing challenge. Moreover, there is increasing evidence that risk of relapse and overall survival can be predicted by assessment of kinetics and depth of response following front-line therapy and monitoring of the leukemic burden using molecular or immunological approaches to minimal residual disease (MRD) detection. These advances present the exciting prospect that panels of pre-treatment parameters affording independent prognostic information can be integrated with precise measurement of treatment response using MRD technologies to provide greater refinement in risk-adapted management of AML. This could lead to further improvements in outcome and serve to identify in a more reliable fashion those patients most likely to benefit from allogeneic transplant in first remission. PMID- 20008225 TI - Optimal induction and post-remission therapy for AML in first remission. AB - Approximately 300,000 patients in the world are diagnosed annually with acute myeloid leukemia (AML). The median age at presentation has been steadily increasing over the past few decades and now is approaching 70 years. Although considerable progress has been made over the past 3 decades in the therapy of AML, two thirds of young adults still die of their disease. The therapy of AML, unlike acute lymphoblastic leukemia (ALL), is based on maximally tolerated induction and post-remission therapy, all given within a few months from diagnosis. While complete remission can be achieved in the majority of young patients, ultimate cure of the disease depends on disease eradication through the administration of post-remission therapy. This is most often done with intensive chemotherapy. Harnessing the immunologic effect of graft-versus-leukemia, as in allogeneic transplantation, has further improved the outcome for many patients. Treatment of older adults, representing the majority of patients with AML, remains quite unsatisfactory. While 40% to 50% can achieve a complete remission, less than 10% are long-term survivors, and the cure rate of older patients has only minimally improved over the past three decades. Significant progress in the treatment of this age group is urgently required. New and targeted agents have much promise, but a definitive clinical role for these has yet to be conclusively established. PMID- 20008226 TI - What is the role of reduced-intensity transplantation in the treatment of older patients with AML? AB - Acute myelogenous leukemia (AML), either de novo or arising out of antecedent myelodysplasia, increases with age and is rarely curable by standard treatments used for younger patients. Recent clinical trials using reduced-intensity allogeneic transplantation regimens suggest that a proportion of patients with this disease can be cured, with results comparable to those achieved in younger patients undergoing fully ablative transplant. Although those patients who undergo transplant in a first remission often do well, the vast majority of older patients have not benefited because of the low successful remission achieved with standard therapy, the delay in initiating a donor search, and the lack of significant benefit from transplantation in patients who are not in remission. New approaches to induction, improvements in reduced-intensity regimens, and earlier donor identification will help expand the potential clinical benefit to a larger number of older patients with the disease. PMID- 20008227 TI - ASH evidence-based guidelines: is there a role for second allogeneic transplant after relapse? AB - A 35-year-old male with a FLT3(+) AML underwent allogeneic peripheral blood stem cell transplant using a myeloablative non-total body irradiation (TBI) conditioning regimen from his HLA-matched sibling donor. Following transplantation, he developed grade II acute graft-versus-host disease (GVHD) that resolved with increasing immunosuppression. The medications were subsequently discontinued, and he did not develop any evidence of chronic GVHD. Eighteen months after transplant, while off all immunosuppression, he developed fatigue and a blood count showed circulating blasts consistent with relapse of his disease. Among the various therapeutic questions is whether there is a role for a second allogeneic transplant to treat his disease and if so, at what time, with what conditioning, and with which type of donor. PMID- 20008228 TI - Predicting clinical outcome in CLL: how and why. AB - The clinical course of patients with chronic lymphocytic leukemia (CLL) is heterogeneous, with some patients experiencing rapid disease progression and others living for decades without requiring treatment. Clinical features and molecular/biologic factors such as ZAP-70, immunoglobulin heavy chain (IGHV) gene mutation status, and cytogenetic abnormalities on fluorescent in situ hybridization (FISH) have been found to be robust predictors of treatment-free survival and overall survival among newly diagnosed patients. Beyond their widely recognized value for providing insight into disease biology and utility for stratifying patient risk in clinical trials, these prognostic tools play an important role in the current counseling and management of patients with CLL. Recent studies have focused on how to combine the results of multiple prognostic assays into an integrated risk stratification system and explored how these characteristics influence response to treatment. This chapter reviews the available tools to stratify patient risk and discusses how these tools can be used in routine clinical practice to individualize patient counseling, guide the frequency of follow-up, and inform treatment selection. PMID- 20008229 TI - Monoclonal B-cell lymphocytosis. AB - The incidence and presenting features of chronic lymphocytic leukemia (CLL) have changed significantly over the last century. Routine diagnostic techniques can now detect very low levels of CLL phenotype cells. Monoclonal B-cell lymphocytosis (MBL) is a relatively recent diagnostic category encapsulating individuals with an abnormal B-cell population but not meeting the diagnostic criteria for a B-cell malignancy. This review focuses on CLL-type MBL, which represents the majority of MBL cases identified in diagnostic laboratories. CLL type MBL has a phenotype identical to CLL and shares the same chromosomal abnormalities even at the lowest levels detectable. Recent evidence suggests that the immunoglobulin gene usage plays a key role in whether the abnormal cells will develop in significant numbers. In most cases, CLL-type MBL is a stable condition with only 1% per year among those presenting for clinical attention developing progressive disease requiring treatment, although suppressed immune function may have a more significant impact on outcome. PMID- 20008230 TI - State-of-the-art treatment of chronic lymphocytic leukemia. AB - The last decade has produced rapid progress in the management of chronic lymphocytic leukemia (CLL). Fludarabine, bendamustine and two monoclonal antibodies, alemtuzumab and rituximab, have been approved by the European and/or American regulatory agencies. Several, novel monoclonal antibodies targeting CD20, CD23 or CD40, as well as drugs designed to interfere with proteins regulating the cell cycle, apoptotic machinery, or leukemic microenvironment (eg, flavopiridol, oblimersen, or lenalidomide), are currently being tested in clinical trials. Furthermore, the increased experience with reduced-intensity allogeneic progenitor cell transplantation allows offering this option to physically fit patients. In addition, new prognostic markers that may influence therapeutic decisions have been identified. This review attempts to summarize the current use of these different modalities in CLL therapy. PMID- 20008231 TI - Initial treatment for patients with CML. AB - For adult patients who present with chronic myeloid leukemia (CML) in chronic phase it is now generally agreed that initial treatment should start with the tyrosine kinase inhibitor (TKI) imatinib at 400 mg daily. Five years after starting imatinib about 60% of these patients will be in complete cytogenetic response (CCyR), still taking imatinib; an appreciable proportion of these will have achieved a major molecular response, defined as a 3-log reduction in the level of BCR-ABL1 transcripts in their blood. The patients in CCyR seem to have a very low risk of relapse to chronic phase or of progression to advanced phase. Other patients may be resistant to imatinib or may experience significant side effects that require change of therapy. The best method of monitoring responding patients is to enumerate Philadelphia chromosome-positive marrow metaphases at 3 month intervals until CCyR and to perform RQ-PCR for BCR-ABL1 transcripts at 3 month intervals after starting imatinib. The recommendations for defining "failure" and "sub-optimal response" proposed by the European LeukemiaNet in 2006 have proved to be a major contribution to assessing responses in individual patients and are now being updated. Patients who fail imatinib may respond to second-generation TKIs, but allogeneic stem cell transplantation still plays an important role for eligible patients who fare badly with TKIs. Patients who present in advanced phases of CML should be treated initially with TKI alone or with TKI in conjunction with cytotoxic drugs, but their overall prognosis is likely to be much inferior to that of those presenting in early chronic phase. PMID- 20008232 TI - Mechanisms of resistance to tyrosine kinase inhibitors in chronic myeloid leukemia and recent therapeutic strategies to overcome resistance. AB - Given its relative rarity, it may at first seem surprising that chronic myeloid leukemia (CML) has garnered so much attention over the last decade. Yet, the advances in molecular pathogenesis that have been derived from studying this leukemia have clearly benefited all of oncology. Moreover, the strides in drug design and development that have also ensued around CML have given rise to what others have called a molecular revolution in cancer therapy. While a majority of patients with chronic phase CML (CP-CML) have an excellent durable response to imatinib (Gleevec, Novartis, Basel, Switzerland), a clear minority will unfortunately have signs of primary or secondary resistance to therapy. Significant efforts geared toward understanding the molecular mechanisms of imatinib resistance have yielded valuable insights into the biology of drug trafficking into and out of cells, epigenetic control of cellular processes, alterations in enzymatic structures, and the rational structural-based design of small molecule enzyme inhibitors. This review will describe the efforts at understanding the pathogenesis of imatinib resistance and the molecular rationale for the development of second- and now third-generation therapies for patients with CML. PMID- 20008233 TI - Monitoring disease response to tyrosine kinase inhibitor therapy in CML. AB - The remarkable progress made in the treatment of chronic myeloid leukemia (CML) over the past decade has been accompanied by steady improvements in our capacity to accurately and sensitively monitor response to therapy. After the initial target of therapy, complete cytogenetic response (CCR), is achieved, peripheral blood BCR-ABL transcript levels measured by real-time quantitative reverse transcriptase PCR (RQ-PCR) define the subsequent response targets, major and complete molecular response (MMR and CMR). The majority of patients on first-line imatinib therapy achieve a "safe haven" defined as a confirmed MMR, but 20% to 30% stop imatinib due to intolerance and/or resistance. Many imatinib-resistant patients can be effectively treated with second generation tyrosine kinase inhibitors (TKIs), but the actual drug selected should be based on the resistance profile of each inhibitor, in addition to issues of tolerance and disease phase. The main purpose of monitoring response with cytogenetics and RQ-PCR is to identify patients likely to achieve better long-term outcome if they are switched early to second-line therapy, either another TKI or an allograft. Mutation screening is most valuable in cases of loss of response to imatinib or a second line TKI, but there are other settings where a high yield of mutations may justify regular mutation screening. PMID- 20008234 TI - Molecular biology of Hodgkin lymphoma. AB - Hodgkin lymphoma (HL) is derived from mature B cells and subdivided into classical HL and nodular lymphocyte predominant HL (NLPHL). HL is unique among human B cell lymphomas because of the rarity of the lymphoma cells, the Hodgkin and Reed-Sternberg (HRS) cells in classical HL and the lymphocyte-predominant (LP) cells in NLPHL, which usually account for 0.1% to 10% of the cells in the affected tissues. Moreover, HRS cells are unique in the extent to which they have lost their B cell-typical gene expression pattern. Deregulation of transcription factor networks plays a key role in this reprogramming process. HRS cells show strong constitutive activity of the transcription factor NF-kappaB. Multiple mechanisms likely contribute to this deregulated activation, including signaling through particular receptors and genetic lesions. Inactivating mutations in the TNFAIP3 tumor suppressor gene, encoding a negative regulator of NF-kappaB activity, were recently identified in about 40% of patients with classical HL. HRS cells are latently infected by Epstein-Barr virus in about 40% of patients, and an important role of this virus in HL pathogenesis-in particular for cases in which HRS cells had lost the capacity to express a B-cell receptor due to destructive somatic mutation-was recently substantiated. PMID- 20008235 TI - Standard therapy of advanced Hodgkin lymphoma. AB - ABVD (doxorubicin, bleomycin, vinblastine and dacarbazine) continues to be the standard of care for patients with advanced-stage Hodgkin lymphoma (HL). Consolidation of primary chemotherapy with radiation or autologous stem cell transplantation (ASCT) has not demonstrated an improvement in overall survival in randomized controlled trials. Regimens such as escalated BEACOPP have more acute and late toxicities and survival benefits have yet to be confirmed. Despite effective therapy, ultimately 30% to 40% of patients with advanced HL will relapse. ASCT has become the standard of care for patients with relapsed or refractory HL based on two randomized trials. The optimal salvage chemotherapy and high dose therapy regimen are not known. Similarly, non-ASCT strategies including salvage radiotherapy or non-ASCT chemotherapy strategies have been reported and have a potential role in selected clinical scenarios. This review summarizes recent clinical trial results in the initial treatment of advanced HL and will focus on second-line treatment strategies for patients with relapsed or refractory disease. PMID- 20008236 TI - Novel treatment strategies for patients with relapsed classical Hodgkin lymphoma. AB - Although classical Hodgkin lymphoma (HL) is considered one of the most curable human cancers, the treatment of patients with relapsed and refractory disease, especially those who relapse after autologous stem cell transplantation, remains challenging. Furthermore, because the median age of the patients is in the mid 30s, the impact of early mortality on the number of years lost from productive life is remarkable. Patients with HL whose disease relapses after stem cell transplantation are rarely cured with current treatment modalities. New drugs and novel treatment strategies that are based on our understanding of the disease biology and signaling pathways are needed to improve treatment outcome for these patients. This review will focus on emerging new treatment modalities that are currently under investigation for patients with relapsed classical HL. PMID- 20008237 TI - The 2008 WHO classification of lymphomas: implications for clinical practice and translational research. AB - The 4(th) edition of the WHO Classification of Tumours of Haematopoietic and Lymphoid Tissues published in 2008 builds upon the success of the 2001 3(rd) edition; new entities are defined, and solutions for problematic categories are sought. Recent studies have drawn attention to the biological overlap between classical Hodgkin lymphoma (CHL) and diffuse large B-cell lymphomas (DLBCL). Similarly, there is a greater appreciation of the borderlands between Burkitt lymphoma and DLBCL. Strategies for the management of these borderline lesions are proposed. Additionally, age-specific and site-specific factors play an important role in the definition of several new entities, which also have biological underpinnings. Among the peripheral T-cell lymphomas (PTCL), more precise definitions were introduced for several entities, including anaplastic large cell lymphoma, angioimmunoblastic T-cell lymphoma, enteropathy-associated T-cell lymphoma, and subcutaneous panniculitis-like T-cell lymphoma. Several new variants of primary cutaneous T-cell lymphomas are proposed. Finally, the subclassification and categorization of the most common lymphoma subtypes, follicular lymphoma (FL) and DLBCL, were altered to enhance diagnostic accuracy and aid in clinical management. The 2008 WHO classification also draws attention to early events in lymphomagenesis. These lesions help delineate the earliest steps in neoplastic transformation and generally mandate a conservative therapeutic approach. The 2001 classification was rapidly adopted for clinical trials and successfully served as a common language for scientists comparing genetic and functional data. The modifications made in the 2008 classification are the result of this successful partnership among pathologists, clinicians, and biologists, but are only a stepping stone to the future. PMID- 20008238 TI - The incidence, natural history, biology, and treatment of transformed lymphomas. AB - Treatment of patients with transformed lymphoma presents a significant challenge to the practicing physician. Indeed, the transformation of follicular lymphoma to a more aggressive histology is inherent to the biology of this disease and is often associated with an aggressive clinical course, resulting in a poor prognosis. Recent population-based studies have better defined the incidence of this event, and recent laboratory studies have defined the molecular and immunological processes associated with transformation. These studies will be discussed in this review, as will the treatment options for these patients. PMID- 20008239 TI - Current treatment standards and emerging strategies in mantle cell lymphoma. AB - Mantle cell lymphoma (MCL) is a unique subtype of B-cell non-Hodgkin lymphomas characterized by the chromosomal translocation t(11;14)(q13;q32) and nuclear cyclin D1 overexpression in the vast majority of cases. Most patients present with advanced stage disease, often with extranodal dissemination, and pursue an aggressive clinical course in the majority of cases. Recent improvement has been achieved by the successful introduction of monoclonal antibodies and dose intensified approaches including autologous stem cell transplantation (ASCT) strategies. With the exception of allogeneic hematopoietic stem cell transplantation, current treatment approaches are non-curative and the corresponding survival curves are characterized by a delayed, but continuous decline and a median survival of 4 to 6 years. However, recently a subset (15%) of long-term survivors have been identified with a rather indolent clinical course even after conventional treatment strategies only. Emerging strategies such as proteasome inhibitors, IMIDs, mTOR inhibitors and others are based on the dysregulated control of cell cycle machinery and impaired apoptotic pathways. Monotherapy of these compounds achieves efficacy comparable to conventional chemotherapy in relapsed MCL, and combination strategies are currently being investigated in numerous trials; however, their introduction into clinical practice and current treatment algorithms remains a challenge. PMID- 20008240 TI - How to treat a newly diagnosed young patient with multiple myeloma. AB - Survival rates of young patients with myeloma have increased markedly in the last decade, mainly due to the use of autologous stem cell transplantation (ASCT) and new, highly efficient rescue treatments. In order to improve the survival of newly diagnosed young patients further, the next steps need to focus on increasing the activity of upfront or debulking regimens, improving the efficacy of ASCT, mainly through the conditioning regimen, and increasing the duration of responses through more effective maintenance or consolidation therapies. Nevertheless, this approach is being challenged by the favorable results obtained with long-term treatment with novel agents and the possibility of reserving the ASCT until relapse. Allogeneic transplantation in newly diagnosed patients should be considered as an investigational procedure and used only in well-designed clinical trials. This review covers the new strategies that are currently under investigation with the aim of optimizing the outcome for newly diagnosed young patients with myeloma. PMID- 20008241 TI - How to treat elderly patients with multiple myeloma: combination of therapy or sequencing. AB - Patients with multiple myeloma aged older than 65 years have traditionally received an oral regimen combining melphalan and prednisone (MP). The introduction of novel agents, such as immunomodulatory drugs and proteasome inhibitors, has substantially changed the treatment paradigm of this disease. Five randomized phase III studies, comparing MP plus thalidomide (MPT) versus MP, have shown that MPT increased time to progression (TTP); however, only two of these five studies showed improvement in overall survival (OS). One randomized study has shown that MP plus bortezomib (MPV) increases both TTP and OS compared with MP. Both MPT and MPV are now regarded as the new standards of care for elderly patients. Other promising results have been reported with MP plus lenalidomide or lenalidomide plus dexamethasone, or the combination of cyclophosphamide, thalidomide, and dexamethasone. Reduced-intensity transplantation can be an option for some patients, especially when novel agents are incorporated into pre-transplant induction and post-transplant consolidation. For patients aged older than 75 years a gentler approach should be used, and doses of standard MPT or MPV should be reduced. An accurate management of treatment-related adverse events with prompt dose-reduction can greatly reduce the rate of early discontinuation and significantly improve treatment efficacy. The choice of treatment should be tailored according to the patient's biologic age and comorbidities, and the expected toxicity profile of the regimen. PMID- 20008242 TI - Novel therapies for relapsed myeloma. AB - Treatment of myeloma relapse needs to be individualized to reflect the effectiveness and toxicities of prior therapies, with consideration given to pragmatic issues such as the tempo of relapse, age of the patient, access to drugs and patient preference. In general, combination therapies have been associated with higher response rates and improved progression-free survival and may be preferable when a rapid response is required. Nevertheless, in a slower tempo relapse it is unclear at this juncture whether sequencing of drugs or multi agent combinations offer superior overall survival results. Fortunately, active novel agents that offer further possibilities for some myeloma patients have become available in clinical trials. In this review we will describe the various classes of novel drugs being tested and the pros and cons of preclinical testing, and will particularly focus on two agents with single-agent activity in myeloma: carfilzomib, a proteasome inhibitor, and pomalidomide, a member of the immunomodulatory class of drugs. PMID- 20008243 TI - ASH evidence-based guidelines: what is the role of maintenance therapy in the treatment of multiple myeloma? AB - A 51-year-old male was diagnosed with an IgA multiple myeloma (MM) after having back pain for several months. His bone marrow showed 30% involvement with plasma cells and his cytogenetics showed t(4:14). His beta2-microglobulin was 6.5 mg/dL at diagnosis and he had multiple lytic lesions, along with a creatinine of 2.3 mg/dL and significant anemia. Induction therapy with lenalidomide, bortezomib and dexamethasone was used, and he was able to achieve complete remission after 4 cycles of therapy. He then went on to receive high-dose chemotherapy with a single autologous stem cell transplant. He tolerated it well and now comes to discuss follow-up treatment plans. He wants to discuss maintenance therapy. PMID- 20008244 TI - Transplantation in adult ALL. AB - The value of the allogeneic graft-versus-leukemia effect in adult acute lymphoblastic leukemia (ALL) has now been conclusively demonstrated and confirmed. While this is true for adults in all age groups, it may not be the best clinical option for young adults for whom increasingly intensive pediatric protocols are clearly of benefit. On the other hand, there is potentially wider applicability of allogeneic donor transplantation for adults 25 to 45 years old, for whom matched unrelated donors may be as safe and effective as sibling donors, and for the patient older than 45 years for whom reduced-intensity conditioning may be a promising way forward. Since the treatment-related mortality of allogeneic transplantation remains significant, careful selection of patients is mandatory. Patients with the Philadelphia chromosome, those with t(4;11) and those with a complex karyotype remain transplant candidates, and allogeneic transplantation remains the best option for salvage, where achievable, in a remission beyond first. As in childhood ALL minimal residual disease studies may be extremely useful in predicting outcome and, therefore, strategy, but at present there are less definite data in adults. Clinical indications to harness the allogeneic effect will mature as the true value of pediatric protocols in adult patients and the safety and efficacy of a sibling, unrelated and reduced intensity transplant emerge in this disease. PMID- 20008245 TI - Allotransplantation for chronic lymphocytic leukemia. AB - Efforts to develop curative treatment strategies for chronic lymphocytic leukemia (CLL) in recent years have focused on allogeneic stem cell transplantation (alloSCT). The crucial anti-leukemic principle of alloSCT in CLL appears to be the immune-mediated anti-host activities conferred with the graft (graft-versus leukemia effects, GVL). Evidence for GVL in CLL is provided by studies analyzing the kinetics of minimal residual disease on response to immune modulation after transplantation, suggesting that GVL can result in complete and durable suppression of the leukemic clone. AlloSCT from matched related or unrelated donors can overcome the treatment resistance of poor-risk CLL, ie, purine analogue refractory disease and CLL with del 17p-. Even with reduced-intensity conditioning, alloSCT in CLL is associated with significant mortality and morbidity due to graft-versus-host disease, which has to be weighed against the risk of the disease when defining the indication for transplantation. Therefore, it can be regarded as a reasonable treatment option only for eligible patients who fulfill accepted criteria for poor-risk disease. If alloSCT is considered, it should be performed before CLL has advanced to a status of complete refractoriness to assure an optimum chance for a successful outcome. Prospective trials are underway to prove whether allo-SCT can indeed change the natural history of poor-risk CLL. PMID- 20008246 TI - Allogeneic stem cell transplantation in follicular lymphoma: recent progress and controversy. AB - Allogeneic stem cell transplantation (allo HCT) is a curative treatment for follicular lymphoma, but is hampered by a relatively high treatment-related mortality and by difficulties in identifying high-risk groups for whom transplant is warranted. Results with myeloablative conditioning have improved, but the field has shifted largely to reduced-intensity conditioning and non-myeloablative transplantation, though morbidity and mortality are also substantial. Some groups have investigated T cell-depleted transplantation, which results in a low rate of chronic graft-versus-host disease (GVHD) and, in most studies, excellent rates of disease control. Overall, outcome after alloHCT for follicular lymphoma correlates more with disease status, with performance status and with comorbidities than with any particular conditioning regimen used. For patients with chemotherapy-sensitive disease, the treatment-related mortality has stabilized in the 15% to 20% range and, depending on the method of GVHD prophylaxis and the donor type, there is an additional 20% to 60% incidence of chronic GVHD. For patients with chemotherapy-refractory disease, both treatment related mortality and recurrence rates are much higher, but their prognosis is dismal with other treatments and some may be cured, particularly with myeloablative transplants. Ongoing studies focus on improving conditioning regimens, on prevention of disease recurrence and on decreasing chronic GVHD. PMID- 20008247 TI - Pathogenesis and management of essential thrombocythemia. AB - The last four years have seen an explosion in our understanding of the myeloproliferative neoplasms. Important and often unexpected insights into the molecular mechanisms responsible for these disorders have been accompanied by the development of new diagnostic tests and by an improved understanding of the relationship between the different disease entities. This review will focus on recent developments in the pathogenesis and management of essential thrombocythemia with a particular emphasis on its phenotypic overlap with polycythemia vera and primary myelofibrosis. PMID- 20008248 TI - Idiopathic erythrocytosis: a disappearing entity. AB - Erythrocytosis results when there is an increased red cell mass and thus an increased hemoglobin. The causes can be divided into primary intrinsic defects of the erythroid progenitor cell and secondary defects, where factors external to the erythroid compartment are responsible. Both can then be further divided into congenital and acquired categories. Congenital causes include mutations of the erythropoietin receptor and defects of the oxygen-sensing pathway including VHL, PHD2 and HIF2A mutations. When fully investigated there remain a number of patients in whom no cause can be elucidated who are currently described as having idiopathic erythrocytosis. Investigation should start with a full history and examination. Having eliminated the common entity polycythemia vera, further direction for investigation is guided by the erythropoietin level. Clinical consequences of the various erythrocytoses are not clear, but in some groups thromboembolic events have been described in young patients. Evidence is lacking to define best management, but aspirin and venesection to a target hematocrit should be considered. PMID- 20008249 TI - Therapeutic potential of JAK2 inhibitors. AB - The discovery of an activating tyrosine kinase mutation JAK2V617F in myeloproliferative neoplasms (MPNs), polycythemia vera (PV), essential thrombocythemia (ET) and primary myelofibrosis (PMF) has resulted in the development of JAK2 inhibitors, of which several are being evaluated in phase I/II clinical studies. It is important to recognize that because the V617F mutation is localized in a region outside the adenosine triphosphate (ATP) binding pocket of JAK2 enzyme, ATP-competitive inhibitors of JAK2 kinase (like the current JAK2 inhibitors in the clinic) are not likely to discriminate between wild-type and mutant JAK2 enzymes. Therefore, JAK2 inhibitors, by virtue of their near equipotent activity against wild-type JAK2 that is important for normal hematopoiesis, may have adverse myelosuppression as an expected side effect, if administered at doses that aim to completely inhibit the mutant JAK2 enzyme. While they may prove to be effective in controlling hyperproliferation of hematopoietic cells in PV and ET, they may not be able to eliminate mutant clones. On the other hand, JAK inhibitors may have great therapeutic benefit by controlling the disease for patients with MPNs who suffer from debilitating signs (eg, splenomegaly) or constitutional symptoms (which presumably result from high levels of circulating cytokines that signal through JAK enzymes). Indeed, the primary clinical benefits observed so far in MF patients have been significant reduction is splenomegaly, elimination of debilitating disease-related symptoms, and weight gain. Most importantly, patients with and without the JAK2V617F mutation appear to benefit to the same extent. In this review we summarize current clinical experience with JAK2 inhibitors in MPNs. PMID- 20008250 TI - The changing classification of myelodysplastic syndromes: what's in a name? AB - What is the most sensible way of organizing the disorderly spectrum of acquired marrow failure states collectively known as the myelodysplastic syndromes (MDS)? While the 2008 version of the World Health Organization classification is the current standard, the recent proliferation of MDS prognostic tools illustrates the usefulness of supplemental information for clinical purposes. Many cases of acquired bone marrow failure do not fit cleanly into established MDS categories, yet an alternative diagnosis is not apparent. The term "idiopathic cytopenias of undetermined significance" (ICUS) has been proposed to describe these cases, but there is a paucity of information about the natural history of ICUS. New data on the natural history of MDS associated with a broad range of cytogenetic abnormalities that were not included in the International Prognostic Scoring System (IPSS), as well as the emerging picture of karyotypically occult DNA changes, promise to inform future classifications. PMID- 20008251 TI - Treatment of MDS: something old, something new, something borrowed... AB - As opposed to the treatment landscape for myelodysplastic syndromes (MDS) two decades ago, potential therapies now abound for the treatment of lower-risk and higher-risk populations. In lower-risk patients, decision tools can be used to determine the likelihood of response to erythropoiesis stimulating agents (ESAs), which have demonstrated survival advantages in retrospective studies in patients with MDS, and whether these patients should be treated initially with ESAs or non growth factor ("active") therapies. Lenalidomide has shown good activity in transfusion-dependent patients with the del(5q) cytogenetic abnormality and modest activity in other lower-risk patients. In higher-risk patients, the DNA methyltransferase inhibitors produce complete and partial responses in 20% to 30% of patients, and for the first time, the MDS drug azacitidine has demonstrated a survival advantage when compared with conventional therapies. Newer therapies stimulate platelet production and target novel pathways, while a panoply of combination studies are underway or recently completed and that likely represent the next frontier in MDS therapy. PMID- 20008252 TI - Supportive care and chelation therapy in MDS: are we saving lives or just lowering iron? AB - The myelodysplastic syndromes (MDS) are characterized by cytopenias and risk of transformation to acute myeloid leukemia (AML). Although new treatments are available, a mainstay in MDS remains supportive care, which aims to minimize the impact of cytopenias and transfusion of blood products. Red blood cell (RBC) transfusions place patients at risk of iron overload (IOL). In beta-thalassemia major (BTM), IOL from chronic RBC transfusions inevitably leads to organ dysfunction and death. With iron chelation therapy (ICT), survival in BTM improved from the second decade to near normal and correlated with ICT compliance. Effects of ICT in BTM include reversal of cardiac arrhythmias, improvement in left ventricular ejection fraction, arrest of hepatic fibrosis, and reduction of glucose intolerance. It is not clear whether these specific outcomes are applicable to MDS. Although retrospective, recent studies in MDS suggest an adverse effect of transfusion dependence and IOL on survival and AML transformation, and that lowering iron minimizes this impact. These data raise important points that warrant further study. ICT is potentially toxic and cumbersome, is costly, and in MDS patients should be initiated only after weighing potential risks against benefits until further data are available to better justify its use. Since most MDS patients eventually require RBC transfusions, the public health implications both of transfusion dependence and ICT in MDS are considerable. This paper summarizes the impact of cytopenias in MDS and treatment approaches to minimize their impact, with a focus on RBC transfusions and their complications, particularly with respect to iron overload. PMID- 20008253 TI - Overview of gene therapy clinical progress including cancer treatment with gene modified T cells. AB - It is now twenty years since the first legal gene transfer studies were approved, and there has been considerable disappointment in the slow rate of progress that followed the initial studies. Gradually, however, as the limitations of available vectors are acknowledged and overcome, and with advances in our understanding of the molecular and cell biology of genetic diseases and of cancer, unequivocal successes are now being reported. In this paper we describe the remaining major roadblocks to successful gene therapy and outline approaches to overcome them. We also illustrate how genetically modified immune system cells are already being used for the effective treatment of hematological and other malignancies, and how these approaches are being modified so that they can be effective in treating a broader range of malignancies. PMID- 20008254 TI - Ten years of gene therapy for primary immune deficiencies. AB - Gene therapy with hematopoietic stem cells (HSC) is an attractive therapeutic strategy for several forms of primary immunodeficiencies. Current approaches are based on ex vivo gene transfer of the therapeutic gene into autologous HSC by vector-mediated gene transfer. In the past decade, substantial progress has been achieved in the treatment of severe combined immundeficiencies (SCID)-X1, adenosine deaminase (ADA)-deficient SCID, and chronic granulomatous disease (CGD). Results of the SCID gene therapy trials have shown long-term restoration of immune competence and clinical benefit in over 30 patients. The inclusion of reduced-dose conditioning in the ADA-SCID has allowed the engraftment of multipotent gene-corrected HSC at substantial level. In the CGD trial significant engraftment and transgene expression were observed, but the therapeutic effect was transient. The occurrence of adverse events related to insertional mutagenesis in the SCID-X1 and CGD trial has highlighted the limitations of current retroviral vector technology. For future applications the risk-benefit evaluation should include the type of vector employed, the disease background and the nature of the transgene. The use of self-inactivating lentiviral vectors will provide significant advantages in terms of natural gene regulation and reduction in the potential for adverse mutagenic events. Following recent advances in preclinical studies, lentiviral vectors are now being translated into new clinical approaches, such as Wiskott-Aldrich Syndrome. PMID- 20008255 TI - Hematopoietic stem cell gene transfer for the treatment of hemoglobin disorders. AB - Hematopoietic stem cell (HSC)-targeted gene transfer is an attractive approach for the treatment of a number of hematopoietic disorders caused by single gene defects. Indeed, in a series of gene transfer trials for two different primary immunodeficiencies beginning early in this decade, outstanding success has been achieved. Despite generally low levels of engrafted, genetically modified HSCs, these trials were successful because of the marked selective advantage of gene corrected lymphoid precursors that allowed reconstitution of the immune system. Unlike the immunodeficiencies, this robust level of in vivo selection is not available to hematopoietic repopulating cells or early progenitor cells following gene transfer of a therapeutic globin gene in the setting of beta-thalassemia and sickle cell disease. Both preclinical and clinical transplant studies involving bone marrow chimeras suggest that 20% or higher levels of engraftment of genetically modified HSCs will be needed for clinical success in the most severe of these disorders. Encouragingly, gene transfer levels in this range have recently been reported in a lentiviral vector gene transfer clinical trial for children with adrenoleukodystrophy. A clinical gene transfer trial for beta thalassemia has begun in France, and one patient with transfusion-dependent HbE/beta-thalassemia has demonstrated a therapeutic effect after transplantation with autologous CD34(+) cells genetically modified with a beta-globin lentiviral vector. Here, the development and recent progress of gene therapy for the hemoglobin disorders is reviewed. PMID- 20008256 TI - Application of semiconductor and metal nanostructures in biology and medicine. AB - Advances in nanotechnology research have led to the creation of new generation of contrast agents, therapeutics, and delivery systems. These applications are expected to significantly improve the diagnosis and treatment of a variety of diseases. Two nanotechnologies-semiconductor and metallic nanostructures-are the most advanced in this young field and have been extensively investigated for clinical use. These nanostructures are currently the "model" for the developments of many novel nanostructures. This review describes their chemical design, tunable properties, and utility in medicine. Furthermore, we will describe the current understanding of their toxicity, which could be barriers to their use for human. PMID- 20008257 TI - Dendrimer-based nanoparticles for cancer therapy. AB - Recent work has suggested that nanoparticles in the form of dendrimers may be a keystone in the future of therapeutics. The field of oncology could soon be revolutionized by novel strategies for diagnosis and therapy employing dendrimer based nanotherapeutics. Several aspects of cancer therapy would be involved. Diagnosis using imaging techniques such as MRI will be improved by the incorporation of dendrimers as advanced contrast agents. This might involve novel contrast agents targeted specifically to cancer cells. Dendrimers can also be being applied to a variety of cancer therapies to improve their safety and efficacy. A strategy, somewhat akin to the "Trojan horse," involves targeting anti-metabolite drugs via vitamins or hormones that tumors need for growth. Further applications of dendrimers in photodynamic therapy, boron neutron capture therapy, and gene therapy for cancer are being examined. This presentation will cover the fundamentals of research utilizing dendrimers for cancer diagnosis and therapy. An evaluation of this new technologies will detail what advantage dendrimer based therapeutics might have over conventional cancer drugs. PMID- 20008258 TI - Imaging circulating cells and lymphoid tissues with iron oxide nanoparticles. AB - The use of nanometer-sized iron oxide nanoparticles and micron-sized iron oxide particles as magnetic resonance (MR) contrast agents has garnered a high degree of interest in diverse areas of biology and medicine. Applications such as cell tracking, molecular imaging, gene detection, and lymphography are being explored to provide insight into disease mechanisms, monitor therapeutic efficacy, and facilitate diagnostic imaging. What makes iron oxide so appealing is a number of favorable properties including high detectability by MR, biodegradability and low toxicity. Here we describe the recent progress on the use of magnetic nanoparticles in imaging circulating cells and lymphoid tissues. The study of the lymph system and the biodistribution of various circulating immune cells is important in the diagnosis, prognosis, and treatment of a wide range of diseases and is expected to have a profound effect on patient outcome. PMID- 20008259 TI - Finding the right academic job. AB - The career path in academic medicine is full of exciting opportunities as well as great achievements and discoveries. In this review, we discuss various obstacles that fellows face as they explore career opportunities in academic hematology/oncology. We also share thoughts on what might be considered steps for success in maneuvering through a search for the "right" position. We discuss several aims including how to define career goals early on, keys to success in an academic career, how to look for an academic job, successful points for the interview process, and how to negotiate a job offer. Finally, we discuss some roadblocks and limitations of academic careers and promotions. These limitations, frustrations and roadblocks should not be a deterrent from pursuing such a great role in the scientific field. This is a new generation of scientific discovery, improvement in healthcare and great advances in research. PMID- 20008260 TI - The academic job offer: what to expect and how to negotiate. AB - In contrast to the familiar college, medical school, residency and fellowship application mechanisms, securing the first academic faculty position has no ground rules and no timelines. The open-ended and idiosyncratic nature of securing a job can be frustrating. Negotiating a competitive start-up package when continuing on the faculty of one's training institution can be particularly challenging. Deciding which offer to accept should be determined less by the "best deal" that can be negotiated and more by the short-term and long-term opportunities for career success. PMID- 20008261 TI - Academic promotion and tenure: a user's guide for junior faculty members. AB - Securing a junior faculty position is an important early step in an academic career in hematology. Shortly thereafter one should begin to plan for eventual promotion and possible tenure. The process is not straightforward, as the "rules of the road" regarding academic positions, academic tracks, assessment and evaluation metrics, and timelines vary immensely from one institution to another. It is critically important, therefore, for the new junior faculty member to become knowledgeable about the institutional policies and "culture" regarding this process. This understanding includes the definition of and criteria for achieving tenure, the academic tracks and the policies for advancement on each track, and the process by which the institutional committee responsible for promotion and tenure conducts its activities. Learning the rules and successfully navigating the academic pathway will help ensure success by achieving the desired promotion and the self-satisfaction, prestige, and financial awards that may accompany it. PMID- 20008266 TI - Nitroglycerin-induced preconditioning: interaction with nitrate tolerance. PMID- 20008267 TI - Protein kinases A and C regulate receptor-mediated increases in cAMP in rabbit erythrocytes. AB - Activation of the beta-adrenergic receptor (beta-AR) or the prostacyclin receptor (IPR) results in increases in cAMP and ATP release from erythrocytes. cAMP levels depend on a balance between synthesis via adenylyl cyclase and hydrolysis by phosphodiesterases (PDEs). Previously, we reported that cAMP increases associated with activation of the beta-AR and IPR in rabbit and human erythrocytes are tightly regulated by distinct PDEs. Importantly, inhibitors of these PDEs potentiated both increases in cAMP and ATP release. It has been shown that increases in protein kinase (PK) activity can activate PDE3 and PDE4. Both PKA and PKC are present in the erythrocyte and can phosphorylate and activate these PDEs. Here we investigate the hypothesis that PKA regulates PDE activity associated with the beta-AR and both PKA and PKC regulate the PDE activity associated with the IPR in rabbit erythrocytes. Pretreatment of erythrocytes with the PKA inhibitor, H89 (10 microM), in the presence of the PDE4 inhibitor, rolipram (10 microM), augmented isoproterenol (1 microM)-induced cAMP increases. In contrast, in the presence of the PDE3 inhibitor, cilostazol (10 microM), pretreatment of erythrocytes with either H89 (1 microM) or two chemically dissimilar inhibitors of PKC, calphostin C (1 microM) or GFX109203X (1 microM), potentiated iloprost (1 microM)-induced cAMP increases. Furthermore, pretreatment of erythrocytes with both H89 and GFX109203X in the presence of cilostazol augmented the iloprost-induced increases in cAMP to a greater extent than either PK inhibitor individually. These results support the hypothesis that PDEs associated with receptor-mediated increases in cAMP in rabbit erythrocytes are regulated by kinases specific to the receptor's signaling pathway. PMID- 20008268 TI - Chronic urotensin-II infusion induces diastolic dysfunction and enhances collagen production in rats. AB - The vasoactive peptide urotensin-II (U-II) is likely to play a key causal role in cardiac remodeling that ultimately leads to heart failure. Its contribution, specifically to the development of diastolic dysfunction and the downstream intracellular signaling, however, remains unresolved. This study interrogates the effect of chronic U-II infusion in normal rats on cardiac structure and function. The contribution of Rho kinase (ROCK) signaling to these pathophysiological changes is evaluated in cell culture studies. Chronic high-dose U-II infusion over 4 wk significantly impaired diastolic function in rats on echocardiography derived Doppler indexes, including E-wave deceleration time (vehicle 56.7 +/- 3.3 ms, U-II 118.0 +/- 21.5 ms; P < 0.01) and mitral valve annulus peak early/late diastolic tissue velocity (vehicle 2.01 +/- 0.19 ms, U-II 1.04 +/- 0.25 ms; P < 0.01). A lower dose of U-II infusion (1 nmol.kg(-1).h(-1)) yielded comparable changes. Diastolic dysfunction was accompanied by molecular [significant increases in procollagen-alpha(1)(I) gene expression on real-time PCR] and morphological (increases in total collagen, P < 0.05, and collagen type-I protein deposition, P < 0.001) evidence of left ventricular (LV) fibrosis following high dose U-II infusion. The ROCK inhibitor GSK-576371 (10(-7) to 10(-5) M) elicited concentration-dependent inhibition of U-II (10(-7) M)-stimulated cardiac fibroblast collagen synthesis and cardiac myocyte protein synthesis. Chronic U-II infusion causes diastolic dysfunction, caused by fibrosis of the LV. The in vitro data suggest that this may be in part occurring via a ROCK-dependent pathway. PMID- 20008269 TI - Diabetic conditions promote binding of monocytes to vascular smooth muscle cells and their subsequent differentiation. AB - Diabetes is associated with significantly accelerated rates of atherosclerosis, key features of which include the presence of excessive macrophage-derived foam cells in the subendothelial space. We examined the hypothesis that enhanced monocyte-vascular smooth muscle cell (VSMC) interactions leading to subendothelial monocyte retention and differentiation to macrophages under diabetic conditions may be underlying mechanisms. Human aortic VSMCs (HVSMCs) treated with diabetic stimuli high glucose (HG) or S100B, a ligand of the receptor for advanced glycation end products, exhibited significantly increased binding of THP-1 monocytic cells. Diabetic stimuli increased the expression of the adhesive chemokine fractalkine (FKN) in HVSMCs. Pretreatment of HVSMCs with FKN or monocyte chemoattractant protein-1 (MCP-1) neutralizing antibodies significantly inhibited monocyte-VSMC binding, whereas monocytes treated with FKN showed enhanced binding to VSMC. Mouse aortic VSMCs (MVSMCs) derived from type 2 diabetic db/db mice exhibited significantly increased FKN levels and binding to mouse WEHI78/24 monocytic cells relative to nondiabetic control db/+ cells. The enhanced monocyte binding in db/db cells was abolished by both FKN and MCP-1 antibodies. Endothelium-denuded aortas from db/db mice and streptozotocin-induced diabetic mice also exhibited enhanced FKN expression and monocyte binding, relative to respective controls. Coculture with HVSMCs increased CD36 expression in THP-1 cells, and this was significantly augmented by treatment of HVSMCs with S100B or HG. CD36 mRNA and protein levels were also significantly increased in WEHI78/24 cells after coincubation with db/db MVSMCs relative to control MVSMCs. These results demonstrate that diabetic conditions may accelerate atherosclerosis by inducing key chemokines in the vasculature that promote VSMC-monocyte interactions, subendothelial monocyte retention, and differentiation. PMID- 20008270 TI - Baroreflex control of muscle sympathetic nerve activity: a nonpharmacological measure of baroreflex sensitivity. AB - The sensitivity of baroreflex control of sympathetic nerve activity (SNA) represents the responsiveness of SNA to changes in blood pressure. In a slightly different analysis, the baroreflex threshold measures the probability of whether a sympathetic burst will occur at a given diastolic blood pressure. We hypothesized that baroreflex threshold analysis could be used to estimate the sensitivity of the sympathetic baroreflex measured by the pharmacological modified Oxford test. We compared four measures of sympathetic baroreflex sensitivity in 25 young healthy participants: the "gold standard" modified Oxford analysis (nitroprusside and phenylephrine), nonbinned spontaneous baroreflex analysis, binned spontaneous baroreflex analysis, and threshold analysis. The latter three were performed during a quiet baseline period before pharmacological intervention. The modified Oxford baroreflex sensitivity was significantly related to the threshold slope (r = 0.71, P < 0.05) but not to the binned (1 mmHg bins) and the nonbinned spontaneous baroreflex sensitivity (r = 0.22 and 0.36, respectively, P > 0.05), which included burst area. The threshold analysis was also performed during the modified Oxford manipulation. Interestingly, we found that the threshold analysis results were not altered by the vasoactive drugs infused for the modified Oxford. We conclude that the noninvasive threshold analysis technique can be used as an indicator of muscle SNA baroreflex sensitivity as assessed by the modified Oxford technique. Furthermore, the modified Oxford method does not appear to alter the properties of the baroreflex. PMID- 20008271 TI - Phospholemman and beta-adrenergic stimulation in the heart. AB - Phosphorylation at serine 68 of phospholemman (PLM) in response to beta adrenergic stimulation results in simultaneous inhibition of cardiac Na(+)/Ca(2+) exchanger NCX1 and relief of inhibition of Na(+)-K(+)-ATPase. The role of PLM in mediating beta-adrenergic effects on in vivo cardiac function was investigated with congenic PLM-knockout (KO) mice. Echocardiography showed similar ejection fraction between wild-type (WT) and PLM-KO hearts. Cardiac catheterization demonstrated higher baseline contractility (+dP/dt) but similar relaxation ( dP/dt) in PLM-KO mice. In response to isoproterenol (Iso), maximal +dP/dt was similar but maximal -dP/dt was reduced in PLM-KO mice. Dose-response curves to Iso (0.5-25 ng) for WT and PLM-KO hearts were superimposable. Maximal +dP/dt was reached 1-2 min after Iso addition and declined with time in WT but not PLM-KO hearts. In isolated myocytes paced at 2 Hz. contraction and intracellular Ca(2+) concentration ([Ca(2+)](i)) transient amplitudes and [Na(+)](i) reached maximum 2 4 min after Iso addition, followed by decline in WT but not PLM-KO myocytes. Reducing pacing frequency to 0.5 Hz resulted in much smaller increases in [Na(+)](i) and no decline in contraction and [Ca(2+)](i) transient amplitudes with time in Iso-stimulated WT and PLM-KO myocytes. Although baseline Na(+)-K(+) ATPase current was 41% higher in PLM-KO myocytes because of increased alpha(1)- but not alpha(2)-subunit activity, resting [Na(+)](i) was similar between quiescent WT and PLM-KO myocytes. Iso increased alpha(1)-subunit current (I(alpha1)) by 73% in WT but had no effect in PLM-KO myocytes. Iso did not affect alpha(2)-subunit current (I(alpha2)) in WT and PLM-KO myocytes. In both WT and NCX1-KO hearts, PLM coimmunoprecipitated with Na(+)-K(+)-ATPase alpha(1)- and alpha(2)-subunits, indicating that association of PLM with Na(+)-K(+)-ATPase did not require NCX1. We conclude that under stressful conditions in which [Na(+)](i) was high, beta-adrenergic agonist-mediated phosphorylation of PLM resulted in time-dependent reduction in inotropy due to relief of inhibition of Na(+)-K(+) ATPase. PMID- 20008272 TI - The arterial reservoir pressure increases with aging and is the major determinant of the aortic augmentation index. AB - The augmentation index predicts cardiovascular mortality and is usually explained as a distally reflected wave adding to the forward wave generated by systole. We propose that the capacitative properties of the aorta (the arterial reservoir) also contribute significantly to the augmentation index and have calculated the contribution of the arterial reservoir, independently of wave reflection, and assessed how these contributions change with aging. In 15 subjects (aged 53 +/- 10 yr), we measured pressure and Doppler velocity simultaneously in the proximal aorta using intra-arterial wires. We calculated the components of augmentation pressure in two ways: 1) into forward and backward (reflected) components by established separation methods, and 2) using an approach that accounts for an additional reservoir component. When the reservoir was ignored, augmentation pressure (22.7 +/- 13.9 mmHg) comprised a small forward wave (peak pressure = 6.5 +/- 9.4 mmHg) and a larger backward wave (peak pressure = 16.2 +/- 7.6 mmHg). After we took account of the reservoir, the contribution to augmentation pressure of the backward wave was reduced by 64% to 5.8 +/- 4.4 mmHg (P < 0.001), forward pressure was negligible, and reservoir pressure was the largest component (peak pressure = 19.8 +/- 9.3 mmHg). With age, reservoir pressure increased progressively (9.9 mmHg/decade, r = 0.69, P < 0.001). In conclusion, the augmentation index is principally determined by aortic reservoir function and other elastic arteries and only to a minor extent by reflected waves. Reservoir function rather than wave reflection changes markedly with aging, which accounts for the age-related changes in the aortic pressure waveform. PMID- 20008273 TI - Involvement of calcium-calmodulin-dependent protein kinase II in endothelin receptor expression in rat cerebral arteries. AB - Experimental cerebral ischemia and organ culture of cerebral arteries result in the enhanced expression of endothelin ET(B) receptors in smooth muscle cells via increased transcription. The present study was designed to evaluate the involvement of calcium-calmodulin-dependent protein kinase (CAMK) in the transcriptional expression of endothelin receptors after organ culture. Rat basilar arteries were incubated for 24 h with or without the CAMK inhibitor KN93 or ERK1/2 inhibitor U0126. The contractile responses to endothelin-1 (ET-1; ET(A) and ET(B) receptor agonist) and sarafotoxin 6c (S6c; ET(B) receptor agonist) were studied using a sensitive myograph. The mRNA levels of the ET(A) and ET(B) receptors and CAMKII were determined by real-time PCR, and their protein levels were evaluated by immunohistochemistry and Western blot. The mRNA levels of CAMKII and the ET(B) receptor increased during organ culture, but there was no change in the expression of the ET(A) receptor. This effect was abolished by coincubation with KN93 or U0126. In functional studies, both inhibitors attenuated the S6c-induced contraction. Incubating the arteries with KN93, but not U0126, decreased the amount of phosphorylated CAMKII. The inhibitors had no effect on the levels of myosin light chain during organ culture, as measured by Western blot. CAMKII is involved in the upregulation of the endothelin ET(B) receptor and interacts with the ERK1/2 pathway to enhance receptor expression. CAMKII has no effect on the contractile apparatus in rat cerebral arteries. PMID- 20008274 TI - Gene expression profiling of dilated cardiomyopathy in older male EP4 knockout mice. AB - Using a line of mice with cardiac-specific knockout (KO) of the EP4 receptor gene, experiments were designed to determine whether a cardiac phenotype developed with age. Cardiac function was assessed by echocardiography in 23- to 33-wk-old male and female KO and littermate controls (WT) mice. After echocardiography, hearts were removed to assess weight, and then some were further processed for histology [myocyte cross-sectional area (MCSA), interstitial collagen fraction (ICF), and macrophage infiltration] and some for extraction of total RNA and protein. Older male KO mice had reduced ejection fraction (EF) coupled with left ventricular dilatation. MCSA and infiltrating macrophages were not different between groups, but ICF increased by 39% in KO mice. In contrast to male KO mice, 30- to 32-wk-old female KO mice had only a slight reduction in EF. To understand gene expression differences between male WT and KO mice, we performed whole genome gene expression profiling (Illumina BeadChips) on hearts of 30-to 32-wk-old mice. Data indicated that 156 genes were overexpressed in the KO hearts more than twofold, including genes involved in remodeling, inflammation, and oxidative stress. Overexpressed chemokines/cytokines were further examined in hearts of 10- to 12-wk-old male KO mice, and we found that growth differentiation factor-15 (GDF-15) expression was higher in KO than in WT hearts. In conclusion, EP4 knockdown in cardiac myocytes in aged male KO mice is in part associated with increased fibrosis, reduced EF, and dilated cardiomyopathy. Early overexpression of GDF-15 in hearts of male KO mice may contribute to or be a marker of the disease phenotype. The absence of serious cardiac dysfunction in aged female mice suggests a sexual dimorphism in the phenotype. PMID- 20008275 TI - Autophagy and protein kinase C are required for cardioprotection by sulfaphenazole. AB - Previously, we showed that sulfaphenazole (SUL), an antimicrobial agent that is a potent inhibitor of cytochrome P4502C9, is protective against ischemia reperfusion (I/R) injury (Ref. 15). The mechanism, however, underlying this cardioprotection, is largely unknown. With evidence that activation of autophagy is protective against simulated I/R in HL-1 cells, and evidence that autophagy is upregulated in preconditioned hearts, we hypothesized that SUL-mediated cardioprotection might resemble ischemic preconditioning with respect to activation of protein kinase C and autophagy. We used the Langendorff model of global ischemia to assess the role of autophagy and protein kinase C in myocardial protection by SUL during I/R. We show that SUL enhanced recovery of function, reduced creatine kinase release, decreased infarct size, and induced autophagy. SUL also triggered PKC translocation, whereas inhibition of PKC with chelerythrine blocked the activation of autophagy in adult rat cardiomyocytes. In the Langendorff model, chelerythrine suppressed autophagy and abolished the protection mediated by SUL. SUL increased autophagy in adult rat cardiomyocytes infected with GFP-LC3 adenovirus, in isolated perfused rat hearts, and in mCherry LC3 transgenic mice. To establish the role of autophagy in cardioprotection, we used the cell-permeable dominant-negative inhibitor of autophagy, Tat Atg5(K130R). Autophagy and cardioprotection were abolished in rat hearts perfused with recombinant Tat-Atg5(K130R). Taken together, these studies indicate that cardioprotection mediated by SUL involves a PKC-dependent induction of autophagy. The findings suggest that autophagy may be a fundamental process that enhances the heart's tolerance to ischemia. PMID- 20008276 TI - Inhibition of soluble epoxide hydrolase preserves cardiomyocytes: role of STAT3 signaling. AB - Soluble epoxide hydrolase (sEH) metabolizes epoxyeicosatrienoic acids (EETs), primarily 14,15-EET. EETs are derived from arachidonic acid via P-450 epoxygenases and are cardioprotective. We tested the hypothesis that sEH deficiency and pharmacological inhibition elicit tolerance to ischemia via EET mediated STAT3 signaling in vitro and in vivo. In addition, the relevance of single nucleotide polymorphisms (SNPs) of EPHX2 (the gene encoding sEH) on tolerance to oxygen and glucose deprivation and reoxygenation and glucose repletion (OGD/RGR) was assessed in male C57BL?6J (WT) or sEH knockout (sEHKO) cardiomyocytes by using transactivator of transcription (TAT)-mediated transduction with sEH mutant proteins. Cell death and hydrolase activity was lower in Arg287Gln EPHX2 mutants vs. nontransduced controls. Excess 14,15-EET and SEH inhibition did not improve cell survival in Arg287Gln mutants. In WT cells, the putative EET receptor antagonist, 14,15-EEZE, abolished the effect of 14,15 EET and sEH inhibition. Cotreatment with 14,15-EET and SEH inhibition did not provide increased protection. In vitro, STAT3 inhibition blocked 14,15-EET cytoprotection, but not the effect of SEH inhibition. However, STAT3 small interfering RNA (siRNA) abolished cytoprotection by 14,15-EET and sEH inhibition, but cells pretreated with JAK2 siRNA remained protected. In vivo, STAT3 inhibition abolished 14,15-EET-mediated infarct size reduction. In summary, the Arg287Gln mutation is associated with improved tolerance against ischemia in vitro, and inhibition of sEH preserves cardiomyocyte viability following OGD/RGR via an EET-dependent mechanism. In vivo and in vitro, 14,15-EET-mediated protection is mediated in part by STAT3. PMID- 20008277 TI - Age-dependent alterations in fibrillar collagen content and myocardial diastolic function: role of SPARC in post-synthetic procollagen processing. AB - Advanced age, independent of concurrent cardiovascular disease, can be associated with increased extracellular matrix (ECM) fibrillar collagen content and abnormal diastolic function. However, the mechanisms causing this left ventricular (LV) remodeling remain incompletely defined. We hypothesized that one determinant of age-dependent remodeling is a change in the extent to which newly synthesized procollagen is processed into mature collagen fibrils. We further hypothesized that secreted protein acidic and rich in cysteine (SPARC) plays a key role in the changes in post-synthetic procollagen processing that occur in the aged myocardium. Young (3 mo old) and old (18-24 mo old) wild-type (WT) and SPARC-null mice were studied. LV collagen content was measured histologically by collagen volume fraction, collagen composition was measured by hydroxyproline assay as soluble collagen (1 M NaCl extractable) versus insoluble collagen (mature cross linked), and collagen morphological structure was examined by scanning electron microscopy. SPARC expression was measured by immunoblot analysis. LV and myocardial structure and function were assessed using echocardiographic and papillary muscle experiments. In WT mice, advanced age increased SPARC expression, myocardial diastolic stiffness, fibrillar collagen content, and insoluble collagen. In SPARC-null mice, advanced age also increased myocardial diastolic stiffness, fibrillar collagen content, and insoluble collagen but significantly less than those seen in WT old mice. As a result, insoluble collagen and myocardial diastolic stiffness were lower in old SPARC-null mice (1.36 +/- 0.08 mg hydroxyproline/g dry wt and 0.04 +/- 0.005) than in old WT mice (1.70 +/- 0.10 mg hydroxyproline/g dry wt and 0.07 +/- 0.005, P < 0.05). In conclusion, the absence of SPARC reduced age-dependent alterations in ECM fibrillar collagen and diastolic function. These data support the hypothesis that SPARC plays a key role in post-synthetic procollagen processing and contributes to the increase in collagen content found in the aged myocardium. PMID- 20008278 TI - Resveratrol, an activator of SIRT1, upregulates sarcoplasmic calcium ATPase and improves cardiac function in diabetic cardiomyopathy. AB - Reduced sarcoplasmic calcium ATPase (SERCA2a) expression has been shown to play a significant role in the cardiac dysfunction in diabetic cardiomyopathy. The mechanism of SERCA2a repression is, however, not known. This study was designed to examine the effect of resveratrol (RSV), a potent activator of SIRT1, on cardiac function and SERCA2a expression in chronic type 1 diabetes. Adult male mice were injected with streptozotocin (STZ) and fed with either a regular diet or a diet enriched with RSV. STZ administration produced progressive decline in cardiac function, associated with markedly reduced SERCA2a and SIRT1 protein levels and increased collagen deposition; RSV treatment to these mice had a tremendous beneficial effect both in terms of improving SERCA2a expression and on cardiac function. In cultured cardiomyocytes, RSV restored SERCA2 promoter activity, which was otherwise highly repressed in high-glucose media. Protective effects of RSV were found to be dependent on its ability to activate Silent information regulator (SIRT) 1. In cardiomyocytes, overexpression of SIRT1 was found sufficient to activate SERCA2 promoter in a dose-dependent manner. In contrast, pretreatment of cardiomyocytes with SIRT1 antagonist, splitomycin, blocked these beneficial effects of RSV. In addition, SIRT1 knockout (+/-) mice were also found to be more sensitive to STZ-induced decline in SERCA2a mRNA. The data demonstrate that, in chronic diabetes, 1) the enzymatic activity of cardiac SIRT1 is reduced, which contributes to reduced expression of SERCA2a and 2) through activation of SIRT1, RSV enhances expression of SERCA2a and improves cardiac function. PMID- 20008279 TI - Project MATCH: training for a promotora intervention. AB - PURPOSE: Adequate training and support are critical for community health workers (promotoras de salud in Spanish) to work effectively. Current literature on promotora training is limited by a focus on promotoras' knowledge and satisfaction immediately after training. The relevance of training to subsequent work performance and the need for ongoing training are rarely addressed. This article describes the training and evaluation components of a promotora intervention focused on diabetes self-management. Training Methods Project MATCH (the Mexican American Trial of Community Health Workers) is a clinical trial designed to test the effectiveness of an intensive, promotora-based intervention to improve disease self-management for Mexican Americans with diabetes. The MATCH investigators designed a multicomponent promotora training program that provided both initial and ongoing training. The investigators used multiple methods to determine promotoras' knowledge levels, initial competency in intervention delivery, and changes in this competency over time. Evaluation Methods and Results The evaluation results show that although the initial training provided a solid knowledge and skills base for the promotoras, the ongoing training was critical in helping them deal with both intervention-related and personal challenges. CONCLUSIONS: The experiences of the MATCH study suggest that in addition to strong initial training, promotora interventions benefit from ongoing training and evaluation to ensure success. PMID- 20008280 TI - Reversible or irreversible remodeling in pulmonary arterial hypertension. AB - Vascular remodeling is an important pathological feature of pulmonary arterial hypertension (PAH), which leads to increased pulmonary vascular resistance, with marked proliferation of pulmonary artery smooth muscle cells (SMC) and/or endothelial cells (EC). Successful treatment of experimental PAH with a platelet derived growth factor (PDGF) receptor tyrosine kinase inhibitor offers the perspective of "reverse remodeling" (i.e., the regression of established pulmonary vascular lesions). Here we ask the question: which forms of pulmonary vascular remodeling are reversible and can such remodeling caused by angiogenic proliferation of EC be reversed? It is important to emphasize that the report showing reduction of vascular remodeling by PDGF receptor tyrosine kinase inhibitor showed only a reduction of the pulmonary artery muscularization in chronic hypoxia and monocrotaline models, which lack the feature of clustered proliferated EC in the lumen of pulmonary arteries. The regression of vascular muscularization is an important manifestation, whereby proliferative adult SMC convert back to a nonproliferative state. In contrast, in vitro experiments assessing the contribution of EC to the development of PAH demonstrated that phenotypically altered EC generated as a consequence of a vascular endothelial growth factor receptor blockade did not reverse to normal EC. Whereas it is suggested that the proliferative state of SMC may be reversible, it remains unknown whether phenotypically altered EC can switch back to a normal monolayer forming EC. This article reviews the pathogenetic concepts of severe PAH and explains the many forms in PAH with reversible or irreversible remodeling. PMID- 20008281 TI - Dynamics of human complement-mediated killing of Klebsiella pneumoniae. AB - With an in vitro system that used a luminescent strain of Klebsiella pneumoniae to assess bacterial metabolic activity in near-real-time, we investigated the dynamics of complement-mediated attack in healthy individuals and in patients presenting to the emergency department with community-acquired severe sepsis. A novel mathematical/statistical model was developed to simplify light output trajectories over time into two fitted parameters, the rate of complement activation and the delay from activation to the onset of killing. Using Factor B depleted serum, the alternative pathway was found to be the primary bactericidal effector: In the absence of B, C3 opsonization as measured by flow cytometry did not progress and bacteria proliferated near exponentially. Defects in bacterial killing were easily demonstrable in patients with severe sepsis compared with healthy volunteers. In most patients with sepsis, the rate of activation was higher than in normal subjects but was associated with a prolonged delay between activation and bacterial killing (P < 0.05 for both). Theoretical modeling suggested that this combination of accentuated but delayed function should allow successful bacterial killing but with significantly greater complement activation. The use of luminescent bacteria allowed for the development of a novel and powerful tool for assessing complement immunology for the purposes of mechanistic study and patient evaluation. PMID- 20008283 TI - 17,18-epoxyeicosatetraenoic acid targets PPARgamma and p38 mitogen-activated protein kinase to mediate its anti-inflammatory effects in the lung: role of soluble epoxide hydrolase. AB - This study sought to assess putative pathways involved in the anti-inflammatory effects of 17,18-epoxyeicosatetraenoic acid (17,18-EpETE), as measured by a decrease in the contractile reactivity and Ca(2+) sensitivity of TNF-alpha pretreated human bronchi. Tension measurements performed in the presence of 12-(3 adamantan-1-yl-ureido)-dodecanoic acid (AUDA), a soluble epoxide hydrolase (sEH) specific inhibitor, demonstrated that 17,18-EpETE reduced the reactivity of TNF alpha-pretreated tissues. The overexpression of sEH detected in patients with asthma and TNF-alpha-treated bronchi contributed to the maintenance of hyperresponsiveness in our models, which involved intracellular proinflammatory cascades. The inhibition of peroxisome proliferator-activated receptor (PPAR)gamma by GW9662 abolished 17,18-EpETE + AUDA-mediated anti-inflammatory effects by inducing IkappaBalpha degradation and cytokine synthesis, indicating that PPARgamma is a molecular target of epoxy-eicosanoids. Western blot analysis revealed that 17,18-EpETE pretreatment reversed the phosphorylation of p38 mitogen-activated protein kinase (p38-MAPK) induced by TNF-alpha in human bronchi. The Ca(2+) sensitivity of human bronchial explants was also quantified on beta-escin permeabilized preparations. The presence of SB203580, a p38-MAPK inhibitor, reversed the effect induced by epoxy-eicosanoid in the presence of AUDA on TNF-alpha-triggered Ca(2+) hypersensitivity by increasing the phosphorylation level of PKC Potentiated Inhibitor Protein-17 (CPI-17) regulatory protein. Moreover, PPARgamma ligands, such as rosiglitazone and 17,18-EpETE, decreased the expression of CPI-17, both at the mRNA and protein levels, whereas this effect was countered by GW9662 treatment in TNF-alpha-treated bronchi. These results demonstrate that 17,18-EpETE is a potent regulator of human lung inflammation and concomitant hyperresponsiveness, and may represent a valuable asset against critical inflammatory bronchial disorder. PMID- 20008282 TI - Cigarette smoke induces nucleic-acid oxidation in lung fibroblasts. AB - Oxidative stress is widely proposed as a pathogenic mechanism for chronic obstructive pulmonary disease (COPD), but the molecular pathway connecting oxidative damage to tissue destruction remains to be fully defined. We suggest that reactive oxygen species (ROS) oxidatively damage nucleic acids, and this effect requires multiple repair mechanisms, particularly base excision pathway components 8-oxoguanine-DNA glycosylase (OGG1), endonuclease III homologue 1 (NTH1), and single-strand-selective monofunctional uracil-DNA glycosylase 1 (SMUG1), as well as the nucleic acid-binding protein, Y-box binding protein 1 (YB1). This study was therefore designed to define the levels of nucleic-acid oxidation and expression of genes involved in the repair of COPD and in corresponding models of this disease. We found significant oxidation of nucleic acids localized to alveolar lung fibroblasts, increased levels of OGG1 mRNA expression, and decreased concentrations of NTH1, SMUG1, and YB1 mRNA in lung samples from subjects with very severe COPD compared with little or no COPD. Mice exposed to cigarette smoke exhibited a time-dependent accumulation of nucleic acid oxidation in alveolar fibroblasts, which was associated with an increase in OGG1 and YB1 mRNA concentrations. Similarly, human lung fibroblasts exposed to cigarette smoke extract exhibited ROS-dependent nucleic-acid oxidation. The short interfering RNA (siRNA)-dependent knockdown of OGG1 and YB1 expression increased nucleic-acid oxidation at the basal state and after exposure to cigarette smoke. Together, our results demonstrate ROS-dependent, cigarette smoke-induced nucleic acid oxidation in alveolar fibroblasts, which may play a role in the pathogenesis of emphysema. PMID- 20008284 TI - Glycogen synthase kinase 3beta activation is a prerequisite signal for cytokine production and chemotaxis in human mast cells. AB - In addition to regulating mast cell homeostasis, the activation of KIT following ligation by stem cell factor promotes a diversity of mast cell responses, including cytokine production and chemotaxis. Although we have previously defined a role for the mammalian target of rapamycin complex 1 in these responses, it is clear that other signals are also required for maximal KIT-dependent cytokine production and chemotaxis. In this study, we provide evidence to support a role for glycogen synthase kinase 3beta (GSK3beta) in such regulation in human mast cells (HuMCs). GSK3beta was observed to be constitutively activated in HuMCs. This activity was inhibited by knockdown of GSK3beta protein following transduction of these cells with GSK3beta-targeted shRNA. This resulted in a marked attenuation in the ability of KIT to promote chemotaxis and, in synergy with FcepsilonRI-mediated signaling, cytokine production. GSK3beta regulated KIT dependent mast cell responses independently of mammalian target of rapamycin. However, evidence from the knockdown studies suggested that GSK3beta was required for activation of the MAPKs, p38, and JNK and downstream phosphorylation of the transcription factors, Jun and activating transcription factor 2, in addition to activation of the transcription factor NF-kappaB. These studies provide evidence for a novel prerequisite priming mechanism for KIT-dependent responses regulated by GSK3beta in HuMCs. PMID- 20008285 TI - Listeria monocytogenes-infected human peripheral blood mononuclear cells produce IL-1beta, depending on listeriolysin O and NLRP3. AB - Different NOD-like receptors, including NLRP1, NLRP3, and NLRC4, as well as the recently identified HIN-200 protein, AIM2, form multiprotein complexes called inflammasomes, which mediate caspase-1-dependent processing of pro-IL-1beta. Listeria monocytogenes is an intracellular pathogen that is actively phagocytosed by monocytes/macrophages and subsequently escapes from the phagosome into the host cell cytosol, depending on its pore-forming toxin listeriolysin O (LLO). In this study, we demonstrate that human PBMCs produced mature IL-1beta when infected with wild-type L. monocytogenes or when treated with purified LLO. L. monocytogenes mutants lacking LLO or expressing a noncytolytic LLO as well as the avirulent Listeria innocua induced strongly impaired IL-1beta production. RNA interference and inhibitor experiments in human PBMCs as well as experiments in Nlrp3 and Rip2 knockout bone marrow-derived macrophages demonstrated that the Listeria-induced IL-1beta release was dependent on ASC, caspase-1, and NLRP3, whereas NOD2, Rip2, NLRP1, NLRP6, NLRP12, NLRC4, and AIM2 appeared to be dispensable. We found that L. monocytogenes-induced IL-1beta production was largely dependent on phagosomal acidification and cathepsin B release, whereas purified LLO activated an IL-1beta production independently of these mechanisms. Our results indicate that L. monocytogenes-infected human PBMCs produced IL 1beta, largely depending on an LLO-mediated phagosomal rupture and cathepsin B release, which is sensed by Nlrp3. In addition, an LLO-dependent but cathepsin B independent NLRP3 activation might contribute to some extent to the IL-1beta production in L. monocytogenes-infected cells. PMID- 20008286 TI - CD40 ligand-induced carcinoma cell death: a balance between activation of TNFR associated factor (TRAF) 3-dependent death signals and suppression of TRAF6 dependent survival signals. AB - The effects of CD40 ligation in an epithelial context are complex, with the level of CD40 engagement influencing the physiological outcome. Low levels of CD40 ligation promote cell survival/proliferation, whereas high levels induce growth arrest/apoptosis. The precise form of the CD40 stimulus affects these responses with the most profound effects in carcinoma cells being induced by membrane-bound rather than recombinant soluble CD40L. However, the signaling pathways underlying these differential responses are yet to be fully characterized. We have investigated the mechanistic differences resulting from CD40 engagement by soluble and membrane-bound ligands using a novel adenovirus-delivered CD40L mutated to resist cleavage from the cell membrane in the CD40-positive EJ bladder carcinoma cell line. We have shown that membrane-bound CD40L induces apoptosis by influencing the balance between apoptotic and survival signals. Thus, membrane bound CD40L stabilizes TNFR-associated factor 3 to induce JNK-dependent apoptosis via release of mitochondrial cytochrome c, caspase 9, and effector caspases 3/7. Further, we have shown that this process is dependent on activation of caspase 8. However, there is also a requirement for suppression of TNFR-associated factor 6 mediated PI3K/Akt-dependent survival signals for apoptosis to occur. These data provide mechanistic insights into the consequences of CD40 activation in carcinoma cells and how these might be exploited in the clinical development of CD40-targeted anticancer therapies. PMID- 20008287 TI - Nucleotide oligomerization binding domain-like receptor signaling enhances dendritic cell-mediated cross-priming in vivo. AB - Nucleotide oligomerization binding domain (Nod)-like receptors are critical cytosolic sensors for the recognition of bacterial peptidoglycan. However, their role in the induction of dendritic cell (DC)-mediated cross-priming remains unclear. In this study, we demonstrate that injecting ligands for Nod1 and Nod2 along with Ag into wild-type mice significantly enhanced the cross-priming of Ag specific CD8+ T cells by CD8alpha+ DCs, as assessed from the expansion of IFN gamma+ CD8+ T cells, CTL activity against Ag-pulsed targets, and the rejection of transplanted tumors expressing the cognate Ag. The enhancement of CD8alpha+ DC mediated cross-priming was likely due to the upregulation of Ag cross presentation and of costimulatory molecules. Our findings collectively indicate that Nod1/2 signaling is critical for the optimal induction of DC cross-priming in vivo, which may offer an alternative therapeutic pathway in cancer and hosts refractory to TLR signals or paralyzed by viral evasion strategy. PMID- 20008288 TI - IFN-gamma-dependent activation of macrophages during experimental infections by Mycobacterium ulcerans is impaired by the toxin mycolactone. AB - Buruli ulcer, caused by Mycobacterium ulcerans infections, is a necrotizing skin disease whose pathogenesis is associated with the exotoxin mycolactone. Despite the relevance of this emergent disease, little is known on the immune response against the pathogen. Following the recent demonstration of an intramacrophage growth phase for M. ulcerans, we investigated the biological relevance of IFN gamma and the antimycobacterial mechanisms activated by this cytokine in M. ulcerans-infected macrophages. Three M. ulcerans strains were tested: 5114 (mutant mycolactone-negative, avirulent strain); 94-1327 (intermediate virulence); and 98-912 (high virulence). We show in this study that IFN-gamma is expressed in mouse-infected tissues and that IFN-gamma-deficient mice display increased susceptibility to infection with strains 5114 and, to a lesser extent, 94-1327, but not with the highly virulent strain. Accordingly, IFN-gamma activated cultured macrophages controlled the proliferation of the avirulent and the intermediate virulent strains. Addition of mycolactone purified from strain 98-912 to cultures of IFN-gamma-activated macrophages infected with the mycolactone-negative strain led to a dose-dependent inhibition of the IFN-gamma induced protective mechanisms, involving phagosome maturation/acidification and increased NO production, therefore resulting in increased bacterial burdens. Our findings suggest that the protection mediated by IFN-gamma in M. ulcerans infected macrophages is impaired by the local buildup of mycolactone. PMID- 20008289 TI - Cutting edge: Vascular endothelial growth factor-mediated signaling in human CD45RO+ CD4+ T cells promotes Akt and ERK activation and costimulates IFN-gamma production. AB - In this study, we find that CD45RO+ memory populations of CD4+ T lymphocytes express the vascular endothelial growth factor (VEGF) receptors KDR and Flt-1 at both the mRNA and protein levels. Furthermore, by Western blot analysis, we find that VEGF increases the phosphorylation and activation of ERK and Akt within CD4+CD45RO+ T cells. These VEGF-mediated signaling responses were inhibited by a KDR-specific small interfering RNA in a VEGF receptor-expressing Jurkat T cell line and by SU5416, a pharmacological KDR inhibitor, in CD4+CD45RO+ T cells. We also find that VEGF augments mitogen-induced production of IFN-gamma in a dose dependent manner (p < 0.001) and significantly (p < 0.05) increases directed chemotaxis of this T cell subset. Collectively, our results for the first time define a novel function for VEGF and KDR in CD45RO+ memory T cell responses that are likely of great pathophysiological importance in immunity. PMID- 20008290 TI - CD56highCD16-CD62L- NK cells accumulate in allergic contact dermatitis and contribute to the expression of allergic responses. AB - Allergic contact dermatitis is a common disease caused by an exaggerated T cell mediated immune response to skin-applied haptens. We show in this study that NK cells affect skin immune responses to haptens by releasing type 1 cytokines and inducing keratinocytes apoptosis. Immunohistochemical stainings demonstrated that NK lymphocytes constitute approximately 10% of the inflammatory infiltrate mostly distributed in the superficial dermis and in the epidermis at the site of intense spongiotic changes. More than 90% of NK cells isolated from allergic contact dermatitis skin showed a CD3-CD56(high)CD16- phenotype by FACS analysis. In addition, they uniformly expressed NKG2A, intermediate to high levels of perforin, and the activating receptors, NKG2D, NKp44, and NKp46, but lacked NKp30 and killer Ig-related receptors. Skin NK lymphocytes displayed a CXCR3+CCR6+CCR5+ chemokine receptor asset for homing into inflamed skin, but not CD62L and CCR7 for lymph node homing. When NK cells from nickel-allergic donors were exposed in vitro to the metal, they failed to proliferate, to upregulate CD69, and to release IFN-gamma, thus indicating that NK lymphocytes do not exhibit memory-like properties to haptens. However, IL-2 released by hapten-driven T lymphocytes rapidly induced the release of IFN-gamma by NK cells and promoted the NK-mediated apoptosis of autologous keratinocytes in a hapten-independent manner. Our findings underline the importance of the interaction between innate and adaptive immune mechanisms for amplification of skin allergic responses to haptens and full expression of allergic contact dermatitis. PMID- 20008291 TI - CD137 promotes proliferation and survival of human B cells. AB - CD137 (4-1BB)-mediated costimulation plays an important role in directing the fate of Ag-stimulated T cells and NK cells, yet the role of CD137 in mediating B cell function is unknown. We found that CD137 is expressed in vitro on anti-Ig stimulated peripheral blood B cells and in vivo on tonsillar B cells with an activated phenotype. In vitro CD137 expression is enhanced by CD40 stimulation and IFN-gamma and is inhibited by IL-4, -10, and -21. The expression of CD137 on activated human B cells is functionally relevant because engagement with its ligand at the time of activation stimulates B cell proliferation, enhances B cell survival, and induces secretion of TNF-alpha and -beta. Our study suggests that CD137 costimulation may play a role in defining the fate of Ag-stimulated human B cells. PMID- 20008292 TI - DNAM-1/CD155 interactions promote cytokine and NK cell-mediated suppression of poorly immunogenic melanoma metastases. AB - A role for NK cells in therapeutic intervention for hematologic malignancies, such as acute myeloid leukemia and multiple myeloma, and nonhematologic malignancies, such as melanoma, is becoming more apparent. DNAM-1 is an NK cell receptor whose importance in facilitating activation signals received by NK cells in natural and cytokine-driven responses to tumor metastases in vivo is poorly explored. In this study, we used matched tumor lines expressing a variety of relevant ligands, neutralizing monoclonal Abs, and DNAM-1 gene-targeted mice to determine the relative importance of DNAM-1-ligand interactions in controlling tumor metastases. Our results demonstrate that NK cells require DNAM-1 for natural or cytokine (IL-2, IL-12, or IL-21) suppression of tumor metastases or their variants expressing CD70 or CD80. In contrast, DNAM-1 was dispensable when tumor cells were targets of Ab-dependent cellular cytotoxicity or presented ligands for NKG2D. CD155 appeared to be a key ligand recognized by DNAM-1 in NK cell-mediated suppression of metastases, and DNAM-1-mediated suppression coincided with perforin activity. Overall, these data implied a general role for DNAM-1-CD155 interactions in NK cell-mediated killing of tumors, even in the presence of tumor CD70 or CD80 expression, and further defined the optimal efficacy requirements of cytokines that directly activate NK cells. PMID- 20008293 TI - CD8 T cell-initiated vascular endothelial growth factor expression promotes central nervous system vascular permeability under neuroinflammatory conditions. AB - Dysregulation of the blood-brain barrier (BBB) is a hallmark feature of numerous neurologic disorders as diverse as multiple sclerosis, stroke, epilepsy, viral hemorrhagic fevers, cerebral malaria, and acute hemorrhagic leukoencephalitis. CD8 T cells are one immune cell type that have been implicated in promoting vascular permeability in these conditions. Our laboratory has created a murine model of CD8 T cell-mediated CNS vascular permeability using a variation of the Theiler's murine encephalomyelitis virus system traditionally used to study multiple sclerosis. Previously, we demonstrated that CD8 T cells have the capacity to initiate astrocyte activation, cerebral endothelial cell tight junction protein alterations and CNS vascular permeability through a perforin dependent process. To address the downstream mechanism by which CD8 T cells promote BBB dysregulation, in this study, we assess the role of vascular endothelial growth factor (VEGF) expression in this model. We demonstrate that neuronal expression of VEGF is significantly upregulated prior to, and coinciding with, CNS vascular permeability. Phosphorylation of fetal liver kinase-1 is significantly increased early in this process indicating activation of this receptor. Specific inhibition of neuropilin-1 significantly reduced CNS vascular permeability and fetal liver kinase-1 activation, and preserved levels of the cerebral endothelial cell tight junction protein occludin. Our data demonstrate that CD8 T cells initiate neuronal expression of VEGF in the CNS under neuroinflammatory conditions, and that VEGF may be a viable therapeutic target in neurologic disease characterized by inflammation-induced BBB disruption. PMID- 20008295 TI - Factor H and factor H-related protein 1 bind to human neutrophils via complement receptor 3, mediate attachment to Candida albicans, and enhance neutrophil antimicrobial activity. AB - The host complement system plays an important role in protection against infections. Several human-pathogenic microbes were shown to acquire host complement regulators, such as factor H (CFH), that downregulate complement activation at the microbial surface and protect the pathogens from the opsonic and lytic effects of complement. Because CFH can also bind to host cells, we addressed the role of CFH and CFH-related proteins as adhesion ligands in host pathogen interactions. We show that the CFH family proteins CFH, CFH-like protein 1 (CFHL1), CFH-related protein (CFHR) 1, and CFHR4 long isoform bind to human neutrophil granulocytes and to the opportunistic human-pathogenic yeast Candida albicans. Two major binding sites, one within the N-terminus and one in the C terminus of CFH, were found to mediate binding to neutrophils. Complement receptor 3 (CD11b/CD18; alpha(M)beta2 integrin) was identified as the major cellular receptor on neutrophils for CFH, CFHL1, and CFHR1, but not for CFHR4 long isoform. CFH and CFHR1 supported cell migration. Furthermore, CFH, CFHL1, and CFHR1 increased attachment of neutrophils to C. albicans. Adhesion of neutrophils to plasma-opsonized yeasts was reduced when CFH binding was inhibited by specific Abs or when using CFH-depleted plasma. Yeast-bound CFH and CFHR1 enhanced the generation of reactive oxygen species and the release of the antimicrobial protein lactoferrin by human neutrophils, and resulted in a more efficient killing of the pathogen. Thus, CFH and CFHR1, when bound on the surface of C. albicans, enhance antimicrobial activity of human neutrophils. PMID- 20008294 TI - Vitamin D decreases respiratory syncytial virus induction of NF-kappaB-linked chemokines and cytokines in airway epithelium while maintaining the antiviral state. AB - Epidemiological studies suggest that low vitamin D levels may increase the risk or severity of respiratory viral infections. In this study, we examined the effect of vitamin D on respiratory syncytial virus (RSV)-infected human airway epithelial cells. Airway epithelium converts 25-hydroxyvitamin D3 (storage form) to 1,25-dihydroxyvitamin D3 (active form). Active vitamin D, generated locally in tissues, is important for the nonskeletal actions of vitamin D, including its effects on immune responses. We found that vitamin D induces IkappaBalpha, an NF kappaB inhibitor, in airway epithelium and decreases RSV induction of NF-kappaB driven genes such as IFN-beta and CXCL10. We also found that exposing airway epithelial cells to vitamin D reduced induction of IFN-stimulated proteins with important antiviral activity (e.g., myxovirus resistance A and IFN-stimulated protein of 15 kDa). In contrast to RSV-induced gene expression, vitamin D had no effect on IFN signaling, and isolated IFN induced gene expression. Inhibiting NF kappaB with an adenovirus vector that expressed a nondegradable form of IkappaBalpha mimicked the effects of vitamin D. When the vitamin D receptor was silenced with small interfering RNA, the vitamin D effects were abolished. Most importantly we found that, despite inducing IkappaBalpha and dampening chemokines and IFN-beta, there was no increase in viral mRNA or protein or in viral replication. We conclude that vitamin D decreases the inflammatory response to viral infections in airway epithelium without jeopardizing viral clearance. This suggests that adequate vitamin D levels would contribute to reduced inflammation and less severe disease in RSV-infected individuals. PMID- 20008296 TI - Cutting edge: Plasmacytoid dendritic cells induce IL-10 production in T cells via the Delta-like-4/Notch axis. AB - Proinflammatory Th1 cells can produce large amounts of the immunosuppressive cytokine IL-10, thereby facilitating the self-limitation of inflammatory responses. Recently, we identified the Notch pathway as a main regulator of IL-10 production by Th1 cells. In this study, we show that plasmacytoid dendritic cells (pDCs), by means of their unique high-level expression of the Notch ligand Delta like (Dll)-4, activate the Notch receptor on T cells to induce robust IL-10 production in vitro and in vivo. pDCs display a distinct pattern of Notch ligands compared with conventional dendritic cells, marked by the constitutive expression of Dll-4, the only Notch ligand to induce IL-10 expression in vivo, and Dll-1, while at the same time lacking the expression of Jagged. We provide a new mechanism for IL-10 induction by pDCs underlining the importance of the Dll 4/Notch axis in the regulation of inflammatory T cell responses. PMID- 20008297 TI - ROCK1 functions as a suppressor of inflammatory cell migration by regulating PTEN phosphorylation and stability. AB - Rho kinases belong to a family of serine/threonine kinases whose role in recruitment and migration of inflammatory cells is poorly understood. We show that deficiency of ROCK1 results in increased recruitment and migration of macrophages and neutrophils in vitro and in vivo. Enhanced migration resulting from ROCK1 deficiency is observed despite normal expression of ROCK2 and a significant reduction in overall ROCK activity. ROCK1 directly binds PTEN in response to receptor activation and is essential for PTEN phosphorylation and stability. In the absence of ROCK1, PTEN phosphorylation, stability, and its activity are significantly impaired. Consequently, increased activation of downstream targets of PTEN, including PIP3, AKT, GSK-3beta, and cyclin D1, is observed. Our results reveal ROCK1 as a physiologic regulator of PTEN whose function is to repress excessive recruitment of macrophages and neutrophils during acute inflammation. PMID- 20008298 TI - Phase 2 study of CEP-701, an orally available JAK2 inhibitor, in patients with primary or post-polycythemia vera/essential thrombocythemia myelofibrosis. AB - Few treatment options exist for patients with myelofibrosis (MF), and their survival is significantly shortened. Activating mutation of the JAK2 tyrosine kinase (JAK2(V617F)) is found in approximately 50% of MF patients. CEP-701 is a tyrosine kinase inhibitor that inhibits JAK2 in in vitro and in vivo experiments. We conducted a phase 2 clinical study of CEP-701 in 22 JAK2(V617F)-positive MF patients (80 mg orally twice daily), and 6 (27%) responded by International Working Group criteria (clinical improvement in all cases): reduction in spleen size only (n = 3), transfusion independency (n = 2), and reduction in spleen size with improvement in cytopenias (n = 1). Median time to response was 3 months, and duration of response was more than or equal to 14 months. No improvement was seen in bone marrow fibrosis or JAK2(V617F) allele burden. Phosphorylated STAT3 levels decreased from baseline in responders while on therapy. Eight patients (36%) experienced grade 3 or 4 toxicity, and 6 (27%) required dose reduction. Main side effects were myelosuppression (grade 3 or 4 anemia, 14%; and thrombocytopenia, 23%) and gastrointestinal disturbances (diarrhea, any grade, 72%; grade 3 or 4, 9%; nausea, grade 1 or 2 only, 50%; vomiting, grade 1 or 2 only, 27%). In conclusion, CEP-701 resulted in modest efficacy and mild but frequent gastrointestinal toxicity in MF patients. The study was registered at http://clinicaltrials.gov as NCT00494585. PMID- 20008299 TI - Mutations of an E3 ubiquitin ligase c-Cbl but not TET2 mutations are pathogenic in juvenile myelomonocytic leukemia. AB - Juvenile myelomonocytic leukemia (JMML) is a rare pediatric myeloid neoplasm characterized by excessive proliferation of myelomonocytic cells. When we investigated the presence of recurrent molecular lesions in a cohort of 49 children with JMML, neurofibromatosis phenotype (and thereby NF1 mutation) was present in 2 patients (4%), whereas previously described PTPN11, NRAS, and KRAS mutations were found in 53%, 4%, and 2% of cases, respectively. Consequently, a significant proportion of JMML patients without identifiable pathogenesis prompted our search for other molecular defects. When we applied single nucleotide polymorphism arrays to JMML patients, somatic uniparental disomy 11q was detected in 4 of 49 patients; all of these cases harbored RING finger domain c-Cbl mutations. In total, c-Cbl mutations were detected in 5 (10%) of 49 patients. No mutations were identified in Cbl-b and TET2. c-Cbl and RAS pathway mutations were mutually exclusive. Comparison of clinical phenotypes showed earlier presentation and lower hemoglobin F levels in patients with c-Cbl mutations. Our results indicate that mutations in c-Cbl may represent key molecular lesions in JMML patients without RAS/PTPN11 lesions, suggesting analogous pathogenesis to those observed in chronic myelomonocytic leukemia (CMML) patients. PMID- 20008300 TI - Two routes to leukemic transformation after a JAK2 mutation-positive myeloproliferative neoplasm. AB - Acute myeloid leukemia (AML) may follow a JAK2-positive myeloproliferative neoplasm (MPN), although the mechanisms of disease evolution, often involving loss of mutant JAK2, remain obscure. We studied 16 patients with JAK2-mutant (7 of 16) or JAK2 wild-type (9 of 16) AML after a JAK2-mutant MPN. Primary myelofibrosis or myelofibrotic transformation preceded all 7 JAK2-mutant but only 1 of 9 JAK2 wild-type AMLs (P = .001), implying that JAK2-mutant AML is preceded by mutation(s) that give rise to a "myelofibrosis" phenotype. Loss of the JAK2 mutation by mitotic recombination, gene conversion, or deletion was excluded in all wild-type AMLs. A search for additional mutations identified alterations of RUNX1, WT1, TP53, CBL, NRAS, and TET2, without significant differences between JAK2-mutant and wild-type leukemias. In 4 patients, mutations in TP53, CBL, or TET2 were present in JAK2 wild-type leukemic blasts but absent from the JAK2 mutant MPN. By contrast in a chronic-phase patient, clones harboring mutations in JAK2 or MPL represented the progeny of a shared TET2-mutant ancestral clone. These results indicate that different pathogenetic mechanisms underlie transformation to JAK2 wild-type and JAK2-mutant AML, show that TET2 mutations may be present in a clone distinct from that harboring a JAK2 mutation, and emphasize the clonal heterogeneity of the MPNs. PMID- 20008301 TI - Alterations in the antigen processing-presenting machinery of transformed plasma cells are associated with reduced recognition by CD8+ T cells and characterize the progression of MGUS to multiple myeloma. AB - We hypothesized that progression of monoclonal gammopathy of undetermined significance (MGUS) to multiple myeloma (MM) reflects the escape of transformed plasma cells from T-cell recognition because of impaired antigen processing presenting machinery (APM). We studied plasma cells and CD8(+) T cells from bone marrow of 20 MGUS patients, 20 MM patients, and 10 control patients. Immunofluorescence and flow cytometry revealed significantly different patterns of APM component expression in plasma cells from the 3 groups. Compared with control patients, MM samples had lower expression of proteasome subunits and peptide transporters and greater expression of chaperones, considering both percentages of stained cells and molecular equivalents of soluble fluorochrome. MGUS samples had intermediate percentages of stained cells but molecular equivalents of soluble fluorochrome similar to control patients. Real-time polymerase chain reaction documented that APM changes occurred at the transcriptional level. Cytotoxicity assays demonstrated that MGUS CD8(+) T cells lysed autologous transformed plasma cells more than MM CD8(+) T cells did. MGUS progression correlated directly with calnexin, calreticulin, and tapasin and indirectly with delta, LMP2, and LMP10 expression levels; MM disease status did not correlate with APM levels. APM changes may allow transformed plasma cells to elude immunesurveillance in the MGUS-MM pathogenetic sequence. PMID- 20008302 TI - Lenalidomide plus dexamethasone versus thalidomide plus dexamethasone in newly diagnosed multiple myeloma: a comparative analysis of 411 patients. AB - The objective of this case-control study was to compare the efficacy and toxicity of lenalidomide plus dexamethasone (len/dex) versus thalidomide plus dexamethasone (thal/dex) as initial therapy for newly diagnosed myeloma. We retrospectively studied 411 newly diagnosed patients treated with len/dex (228) or thal/dex (183) at the Mayo Clinic. The differences were similar in a matched pair analysis that adjusted for age, sex, transplantation status, and dexamethasone dose. The proportions of patients achieving at least a partial response to len/dex and thal/dex were 80.3% versus 61.2%, respectively (P < .001); very good partial response rates were 34.2% and 12.0%, respectively (P < .001). Patients receiving len/dex had longer time to progression (median, 27.4 vs 17.2 months; P = .019), progression-free survival (median, 26.7 vs 17.1 months; P = .036), and overall survival (median not reached vs 57.2 months; P = .018). A similar proportion of patients in the 2 groups experienced at least one grade 3 or 4 adverse event (57.5% vs 54.6%, P = .568). Main grade 3 or 4 toxicities of len/dex were hematologic, mainly neutropenia (14.6% vs 0.6%, P < .001); the most common toxicities in thal/dex were venous thromboembolism (15.3% vs 9.2%, P = .058) and peripheral neuropathy (10.4% vs 0.9%, P < .001). Len/dex appears well tolerated and more effective than thal/dex. Randomized trials are needed to confirm these results. PMID- 20008303 TI - Targeting distinct tumor-infiltrating myeloid cells by inhibiting CSF-1 receptor: combating tumor evasion of antiangiogenic therapy. AB - Tumor-infiltrating myeloid cells (TIMs) support tumor growth by promoting angiogenesis and suppressing antitumor immune responses. CSF-1 receptor (CSF1R) signaling is important for the recruitment of CD11b(+)F4/80(+) tumor-associated macrophages (TAMs) and contributes to myeloid cell-mediated angiogenesis. However, the impact of the CSF1R signaling pathway on other TIM subsets, including CD11b(+)Gr-1(+) myeloid-derived suppressor cells (MDSCs), is unknown. Tumor-infiltrating MDSCs have also been shown to contribute to tumor angiogenesis and have recently been implicated in tumor resistance to antiangiogenic therapy, yet their precise involvement in these processes is not well understood. Here, we use the selective pharmacologic inhibitor of CSF1R signaling, GW2580, to demonstrate that CSF-1 regulates the tumor recruitment of CD11b(+)Gr 1(lo)Ly6C(hi) mononuclear MDSCs. Targeting these TIM subsets inhibits tumor angiogenesis associated with reduced expression of proangiogenic and immunosuppressive genes. Combination therapy using GW2580 with an anti-VEGFR-2 antibody synergistically suppresses tumor growth and severely impairs tumor angiogenesis along with reverting at least one TIM-mediated antiangiogenic compensatory mechanism involving MMP-9. These data highlight the importance of CSF1R signaling in the recruitment and function of distinct TIM subsets, including MDSCs, and validate the benefits of targeting CSF1R signaling in combination with antiangiogenic drugs for the treatment of solid cancers. PMID- 20008305 TI - Intraperitoneal VEGF inhibition using bevacizumab: a potential approach for the symptomatic treatment of malignant ascites? AB - Despite overall improvements in oncological care in the palliative setting, symptomatic malignant ascites remains a severe clinical problem. This form of effusion is known to be widely resistant to established modes of systemic therapy. Accordingly, frequent paracentesis often represents the only effective way for symptom relief in patients with advanced cancer. This invasive mode of therapy, however, is often very burdensome for the patient who is already severely distressed by the underlying malignancy. Recently, the trifunctional monoclonal antibody catumaxomab given i.p. has shown symptom relief in patients with ovarian cancer and malignant ascites. On another front, the release of vascular endothelial growth factor (VEGF) by tumor cells has been identified as a main factor promoting the i.p. secretion of fluid. Accordingly, recent evidence suggests that targeting VEGF may have the potential to suspend the ascites production resulting from peritoneal metastasis. Here, we review preclinical and clinical data supporting this hypothesis. We show current evidence suggesting that the i.p. application of the anti-VEGF antibody bevacizumab, which is already in use as an i.v. therapeutic drug for a variety of tumors, might represent an effective way to prevent local fluid accumulation. Because such an effect would result in significant relief for patients, future clinical studies should stringently assess the effectiveness of this targeted therapy for the treatment of malignant i.p. effusions. PMID- 20008306 TI - Is chemotherapy alone for early Hodgkin lymphoma an emerging therapeutic option? PMID- 20008304 TI - Acute T-cell leukemias remain dependent on Notch signaling despite PTEN and INK4A/ARF loss. AB - NOTCH1 is activated by mutation in more than 50% of human T-cell acute lymphoblastic leukemias (T-ALLs) and inhibition of Notch signaling causes cell cycle/growth arrest, providing rationale for NOTCH1 as a therapeutic target. The tumor suppressor phosphatase and tensin homolog (PTEN) is also mutated or lost in up to 20% of cases. It was recently observed among human T-ALL cell lines that PTEN loss correlated with resistance to Notch inhibition, raising concern that patients with PTEN-negative disease may fail Notch inhibitor therapy. As these studies were limited to established cell lines, we addressed this issue using a genetically defined mouse retroviral transduction/bone marrow transplantation model and observed primary murine leukemias to remain dependent on NOTCH1 signaling despite Pten loss, with or without additional deletion of p16(Ink4a)/p19(Arf). We also examined 13 primary human T-ALL samples obtained at diagnosis and found no correlation between PTEN status and resistance to Notch inhibition. Furthermore, we noted in the mouse model that Pten loss accelerated disease onset and produced multiclonal tumors, suggesting NOTCH1 activation and Pten loss may collaborate in leukemia induction. Thus, in contrast to previous findings with established cell lines, these results indicate PTEN loss does not relieve primary T-ALL cells of their "addiction" to Notch signaling. PMID- 20008307 TI - ECG criteria to identify epicardial ventricular tachycardia in nonischemic cardiomyopathy. AB - BACKGROUND: ECG criteria identifying epicardial (EPI) origin for ventricular tachycardia (VT) in nonischemic cardiomyopathy have not been determined. Endocardial (ENDO) and EPI basal left ventricle fibrosis characterizes the VT substrate. METHODS AND RESULTS: We assessed the QRS from 102 basal superior/lateral EPI and 67 comparable ENDO pace maps in 14 patients with nonischemic cardiomyopathy. Pace mapping focused on low bipolar voltage areas. Published morphology criteria: q wave in lead I (QWLI) and no q waves in inferior leads and interval criteria: pseudo-delta wave > or =34 ms, intrinsicoid deflection time > or =85 ms, shortest RS complex > or =121 ms, and maximum deflection index > or =0.55 were assessed for ability to identify EPI origin. Sixteen EPI and 8 ENDO of the 34 mapped VTs (71%) in the study population and 14 EPI and 7 ENDO VTs from an 11-patient validation cohort were localized to basal superior/lateral left ventricle and corroborated pacing data. A QWL1 was seen in EPI but not ENDO pace maps (91% versus 4%; P<0.001), identified 14 of 16 EPI VTs (sensitivity, 88%), and was seen in 1 of 8 ENDO VTs (specificity, 88%). None of the remaining criteria achieved similar sensitivity without specificity <50%. We identified 4 criteria (q waves in inferior leads, pseudo-delta wave > or =75 ms, maximum deflection index > or =0.59, and QWL1) having > or =95% specificity and > or =20% sensitivity in identifying EPI/ENDO origin for pace maps. This 4-step algorithm identified the origin in 109 of 115 pace maps (95%), 21 of 24 VTs (88%) in the study population, and 19 of 21 VTs (90%) in validation cohort. CONCLUSIONS: Morphological ECG features that describe the initial QRS vector can help identify basal-superior/lateral EPI VTs in nonischemic cardiomyopathy. PMID- 20008308 TI - Identifying the effective components of interventions. PMID- 20008309 TI - The first anniversary: stress, well-being, and optimism in older widows. AB - The first anniversary for older widows (n = 47) has been explored during Months 11, 12, and 13. Concurrent correlations show that optimism inversely correlates with psychological (intrusion and avoidance) stress as measured with the Impact of Event Scale (r = -.52 to -.66, p < .005) and positively correlates with well being (physical: r = .36 to .46, p < .025; psychosocial: r = .58 to .72, p < .005; spiritual: r = .50 to .69, p < .005). Lagged correlation patterns suggest that higher levels of optimism at a given time are associated with higher life satisfaction and spiritual well-being at later times. Psychological stress is higher at Month 12 when compared to Month 13, t(43) = 2.54, p = .01, but not when compared to Month 11, t(43) = 1.49, p > .10. There are no significant differences in physiologic stress (salivary cortisol) or well-being during the first anniversary of spousal bereavement. PMID- 20008310 TI - Death worries and quality of life in younger breast cancer survivors. PMID- 20008311 TI - A phenomenological study of nurses' experience of grief following patient death. PMID- 20008312 TI - Differences in pain intensity and quality by pain pattern in outpatients with cancer. PMID- 20008313 TI - A pilot study of African American women's narratives about their newborn feeding choices. PMID- 20008314 TI - The effect of workforce shortages on nurse leader turnover and succession in U.S. hospitals. PMID- 20008315 TI - The impact of health literacy and patient trust on glycemic control. PMID- 20008316 TI - Knowledge of heart attack symptoms and risk factors among native Thais. PMID- 20008317 TI - Validity and reliability of the evidence-based practice self-efficacy scale. PMID- 20008318 TI - Substance abuse and chronic venous disorders. PMID- 20008319 TI - Relationships among sleep duration, sleep quality, fatigue, and depression in fathers of twins. PMID- 20008320 TI - Mitochondrial transcription factor Mtf1 traps the unwound non-template strand to facilitate open complex formation. AB - The catalytic subunit of the mitochondrial (mt) RNA polymerase (RNAP) is highly homologous to the bacteriophage T7/T3 RNAP. Unlike the phage RNAP, however, the mtRNAP relies on accessory proteins to initiate promoter-specific transcription. Rpo41, the catalytic subunit of the Saccharomyces cerevisiae mtRNAP, requires Mtf1 for opening the duplex promoter. To elucidate the role of Mtf1 in promoter specific DNA opening, we have mapped the structural organization of the mtRNAP using site-specific protein-DNA photo-cross-linking studies. Both Mtf1 and Rpo41 cross-linked to distinct sites on the promoter DNA, but the dominant cross-links were those of the Mtf1, which indicates a direct role of Mtf1 in promoter specific binding and initiation. Strikingly, Mtf1 cross-linked with a high efficiency to the melted region of the promoter DNA, based on which we suggest that Mtf1 facilitates DNA melting by trapping the non-template strand in the unwound conformation. Additional strong cross-links of the Mtf1 were observed with the -8 to -10 base-paired region of the promoter. The cross-linking results were incorporated into a structural model of the mtRNAP-DNA, created from a homology model of the C-terminal domain of Rpo41 and the available structure of Mtf1. The promoter DNA is sandwiched between Mtf1 and Rpo41 in the structural model, and Mtf1 closely associates mainly with one face of the promoter across the entire nona-nucleotide consensus sequence. Overall, the studies reveal that in many ways the role of Mtf1 is analogous to the transcription factors of the multisubunit RNAPs, which provides an intriguing link between single- and multisubunit RNAPs. PMID- 20008321 TI - Comparing and ranking hospitals based on outcome: results from The Netherlands Stroke Survey. AB - BACKGROUND: Measuring quality of care and ranking hospitals with outcome measures poses two major methodological challenges: case-mix adjustment and variation that exists by chance. AIM: To compare methods for comparing and ranking hospitals that considers these. METHODS: The Netherlands Stroke Survey was conducted in 10 hospitals in the Netherlands, between October 2002 and May 2003, with prospective and consecutive enrollment of patients with acute brain ischaemia. Poor outcome was defined as death or disability after 1 year (modified Rankin scale of > or =3). We calculated fixed and random hospital effects on poor outcome, unadjusted and adjusted for patient characteristics. We compared the hospitals using the expected rank, a novel statistical measure incorporating the magnitude and the uncertainty of differences in outcome. RESULTS: At 1 year after stroke, 268 of the total 505 patients (53%) had a poor outcome. There were substantial differences in outcome between hospitals in unadjusted analysis (chi(2) = 48, 9 df, P < 0.0001). Adjustment for 12 confounders led to halving of the chi(2) (chi(2) = 24). The same pattern was observed in random effects analysis. Estimated performance of individual hospitals changed considerably between unadjusted and adjusted analysis. Further changes were seen with random effect estimation, especially for smaller hospitals. Ordering by expected rank led to shrinkage of the original ranks of 1-10 towards the median rank of 5.5 and to a different order of the hospitals, compared to ranking based on fixed effects. CONCLUSION: In comparing and ranking hospitals, case-mix-adjusted random effect estimates and the expected ranks are more robust alternatives to traditional fixed effect estimates and simple rankings. PMID- 20008322 TI - IIp45 inhibits cell migration through inhibition of HDAC6. AB - IIp45 (aka MIIP) is a newly discovered gene whose protein product inhibits cell migration. HDAC6 is a class IIb deacetylase that specifically deacetylates alpha tubulin, modulates microtubule dynamics, and promotes cell migration. A yeast two hybrid assay using IIp45 as bait identified HDAC6 protein as a binding partner of IIp45. This physical interaction of the two functionally antagonistic proteins was confirmed by glutathione S-transferase pulldown assay and co immunoprecipitation assay in human cells. Serial deletion constructs of HDAC6 were used to characterize the interaction of HDAC6 and IIp45, and this analysis found that the two catalytic domains of HDAC6 protein are required for IIp45 binding. We examined the protein expression patterns of IIp45 and HDAC6 in glioma tissues. Elevated protein levels of HDAC6 were found in high grade glioma samples, in contrast to the decreased protein expression of IIp45. The potential negative regulation of HDAC6 expression by IIp45 was confirmed in cell lines with altered IIp45 expression by constitutive overexpression or small interfering RNA knockdown. Protein turnover study revealed that overexpression of IIp45 significantly reduces the intracellular protein stability of endogenous HDAC6, indicating a possible mechanism for the negative regulation of HDAC6 by IIp45. Results from the HDAC activity assay demonstrated that overexpressed IIp45 effectively decreases HDAC6 activity, increases acetylated alpha-tubulin, and reduces cell migration. The increased cell migration resulting from siIIp45 knockdown was significantly reversed by co-transfection of siHDAC6. Thus, we report here for the first time a novel mechanism by which IIp45 inhibits cell motility through inhibition of HDAC6. PMID- 20008323 TI - Involvement of the interaction of afadin with ZO-1 in the formation of tight junctions in Madin-Darby canine kidney cells. AB - Tight junctions (TJs) and adherens junctions (AJs) are major junctional apparatuses in epithelial cells. Claudins and junctional adhesion molecules (JAMs) are major cell adhesion molecules (CAMs) at TJs, whereas cadherins and nectins are major CAMs at AJs. Claudins and JAMs are associated with ZO proteins, whereas cadherins are associated with beta- and alpha-catenins, and nectins are associated with afadin. We previously showed that nectins first form cell-cell adhesions where the cadherin-catenin complex is recruited to form AJs, followed by the recruitment of the JAM-ZO and claudin-ZO complexes to the apical side of AJs to form TJs. It is not fully understood how TJ components are recruited to the apical side of AJs. We studied the roles of afadin and ZO-1 in the formation of TJs in Madin-Darby canine kidney (MDCK) cells. Before the formation of TJs, ZO 1 interacted with afadin through the two proline-rich regions of afadin and the SH3 domain of ZO-1. During and after the formation of TJs, ZO-1 dissociated from afadin and associated with JAM-A. Knockdown of afadin impaired the formation of both AJs and TJs in MDCK cells, whereas knockdown of ZO-1 impaired the formation of TJs, but not AJs. Re-expression of full-length afadin restored the formation of both AJs and TJs in afadin-knockdown MDCK cells, whereas re-expression of afadin-DeltaPR1-2, which is incapable of binding to ZO-1, restored the formation of AJs, but not TJs. These results indicate that the transient interaction of afadin with ZO-1 is necessary for the formation of TJs in MDCK cells. PMID- 20008324 TI - Molecular insights into mammalian end-binding protein heterodimerization. AB - Microtubule plus-end tracking proteins (+TIPs) are involved in many microtubule based processes. End binding (EB) proteins constitute a highly conserved family of +TIPs. They play a pivotal role in regulating microtubule dynamics and in the recruitment of diverse +TIPs to growing microtubule plus ends. Here we used a combination of methods to investigate the dimerization properties of the three human EB proteins EB1, EB2, and EB3. Based on Forster resonance energy transfer, we demonstrate that the C-terminal dimerization domains of EBs (EBc) can readily exchange their chains in solution. We further document that EB1c and EB3c preferentially form heterodimers, whereas EB2c does not participate significantly in the formation of heterotypic complexes. Measurements of the reaction thermodynamics and kinetics, homology modeling, and mutagenesis provide details of the molecular determinants of homo- versus heterodimer formation of EBc domains. Fluorescence spectroscopy and nuclear magnetic resonance studies in the presence of the cytoskeleton-associated protein-glycine-rich domains of either CLIP-170 or p150(glued) or of a fragment derived from the adenomatous polyposis coli tumor suppressor protein show that chain exchange of EBc domains can be controlled by binding partners. Extension of these studies of the EBc domains to full-length EBs demonstrate that heterodimer formation between EB1 and EB3, but not between EB2 and the other two EBs, occurs both in vitro and in cells as revealed by live cell imaging. Together, our data provide molecular insights for rationalizing the dominant negative control by C-terminal EB domains and form a basis for understanding the functional role of heterotypic chain exchange by EBs in cells. PMID- 20008326 TI - Timing and duration of European larch growing season along altitudinal gradients in the Swiss Alps. AB - The 2007 European larch (Larix decidua Mill.) growing season was monitored along two elevational transects in the Lotschental valley in the Swiss Alps. Phenological observations and weekly microcore sampling of 28 larch trees were conducted between April and October 2007 at seven study sites regularly spaced from 1350 to 2150 m a.s.l. on northwest- and southeast-facing slopes. The developmental stages of nearly 75,000 individual cells assessed on 1200 thin sections were used to investigate the links between the trees' thermal regimes and growth phases including the beginning and ending of cell enlargement, wall thickening and maturation of the stem wood. Needles appeared approximately 3-4 weeks earlier than stem growth. The duration of ring formation lasted from mid May to the end of October, with the length of the growing season decreasing along elevation from 137 to 101 days. The onset of the different growing seasons changed by 3-4 days per 100 m elevation; the ending of the growing season, however, appeared minimally related to altitude. If associated with the monitored altitudinal lapse rate of -0.5 degrees C per 100 m, these results translate into a lengthening of the growing season by approximately 7 days per degree Celsius. This study provides new data on the timing and duration of basic growth processes and contributes to quantification of the impacts of global warming on tree growth and productivity. PMID- 20008325 TI - The tumor suppressor protein HBP1 is a novel c-myc-binding protein that negatively regulates c-myc transcriptional activity. AB - c-Myc is an important transcription factor that regulates cellular proliferation, cell growth, and differentiation. A number of transcriptional co-factors for c Myc have been described that have binding sites within highly conserved regions of the c-Myc transactivational domain (TAD). Given the importance of the c-Myc TAD, we set out to identify new proteins that interact with this region using a yeast two-hybrid assay. HBP1 was identified in our screen as a protein that interacts with full-length c-Myc but not a c-Myc mutant lacking the TAD. HBP1 is a transcriptional repressor and has been shown to negatively regulate the cell cycle. A correlation between HBP1 under-expression and breast cancer relapse has been described, suggesting that HBP1 may be an important tumor suppressor protein. We have found that HBP1 binds c-Myc in cells, and expression of HBP1 inhibits c-Myc transactivational activity at least partly by preventing c-Myc binding to target gene promoters. c-Myc binds to the C terminus of HBP1, a region lost in some breast tumors, and some HBP1 mutants found in breast cancer weakly interact with and/or no longer negatively regulate c-Myc. This work adds to our understanding of c-Myc regulation and mechanisms of tumor suppression by HBP1. PMID- 20008327 TI - Nitrogen partitioning in orchard-grown Macadamia integrifolia. AB - Nut yield is highly variable in commercial macadamia production, and to ensure that nitrogen (N) supply does not limit yield, high rates of N fertilizer are generally applied. To elucidate N source and sink relations in mature Macadamia integrifolia Maiden et Betche trees, we traced (15)N label after injection into individual branches and, after soil application, analysed xylem sap and examined the effects of hedging on tree N relations. Xylem sap N and sugar composition and concentration changed in relation to phenology and tree management. Canopy position did not affect xylem sap N concentration but sampling date had a significant effect. Hedging in spring was associated with a rapid and dramatic reduction of the concentration of xylem sap N until the following autumn, but unhedged trees were not available to unequivocally assess the significance of the results. Following (15)N-branch injection in winter, most (15)N label was incorporated into flushing leaves and into bark. After (15)N injection in spring, flushing leaves and flowers were most strongly (15)N-labelled. In late spring, (15)N label was equally incorporated by developing nuts that were retained or later abscised. Soil (15)N application in summer resulted in (15)N-labelling of outer and mid-canopy leaves. In the following spring, (15)N label was translocated to flushing leaves, flowers and developing nuts. The results indicate that outer and mid-canopy leaves are the main N sink for soil-derived N during the vegetative phase and a N source for developing tissues during the reproductive phase. Our study provides evidence that N supply to developing nuts is not a primary cause for nut abscission, supporting the notion that high N fertilizer application rates do not improve nut retention. We propose that current orchard design and hedging practices should be reviewed in context of the role of outer canopy leaves as a source of N for reproductive tissues. PMID- 20008328 TI - Left atrial intramural haematoma associated with mitral annular calcification. AB - Left atrial wall haematoma is a very uncommon entity, associated mainly to cardiac surgery, interventional procedures, or trauma. Spontaneous cases are supposed to be associated with left atrial wall pathology. We present a case of a 53-year-old male who was admitted for prolonged chest pain, with transthoracic and transesophagic echocardiography documentation of a left atrial mass in close proximity to a mitral annular calcification. Tissue characterization with cardiac magnetic resonance suggested the aetiology of the mass, which was confirmed histologically. PMID- 20008329 TI - Cardiac sarcoma presenting as heart failure and diagnosed as recurrent myxoma by echocardiogram. AB - We report a case of an extremely rare high-grade, undifferentiated cardiac sarcoma. The patient with left atrial myxoma resected 8 months ago presented with pneumonia, congestive heart failure, and subsequently diagnosed to have cardiac sarcoma. Transoesophageal echocardiogram played an important role in diagnosis of left atrial mass. High index of suspicion is required to diagnose left atrial tumour as initially it can present as pneumonia or congestive heart failure and left atrial tumours are not always the myxoma. PMID- 20008330 TI - Molecular suicide notes: last call from apoptosing cells. AB - Dying cells actively attract professional phagocytes by sending out soluble 'find me' signals. The characterization and identification of the corresponding molecular entities have revealed that 'find-me' signals come in different flavors, including peptides, proteins, lipids and complex structures, such as apoptotic microblebs. With the very recent report by Elliott et al., nucleotides as a novel nuance enter the menu. PMID- 20008331 TI - Neither too much nor too little: reactive oxygen species levels regulate Drosophila hematopoiesis. AB - Owing to their fast diffusion, reactive oxygen species (ROS) are important tissue signalling components. High levels of ROS are generally considered as deleterious to cells. In vivo experiments in the Drosophila hematopoietic organ now challenge this view by showing that high ROS levels are intrinsically required to prime myeloid-like progenitors to differentiate. PMID- 20008332 TI - Destruction of a destructor: a new avenue for cancer therapeutics targeting the Wnt pathway. AB - A new study in Nature has identified a small molecule inhibitor for the oncogenic Wnt/beta-catenin pathway that is responsible for many human cancers. This inhibitor targets an unsuspected cellular enzyme, Tankyrase, which controls the destruction of a beta-catenin destructor. PMID- 20008333 TI - Live show of Rho GTPases in cell migration. AB - Rho GTPases, including RhoA, Rac1 and Cdc42, are key regulators of cell migration in animals. In a recent issue of Nature, two papers present delicate studies on precise roles and spatiotemporal coordination of Rho GTPases in live cells, using either a photoactivatable Rac1 or specific biosensors, together with computational multiplexing method. In addition to implications in cell migration, analytical tools and methods used in these works can also be extended into other live cell studies on complex cellular/molecular events. PMID- 20008334 TI - WD40 repeats arrange histone tails for spreading of silencing. AB - Direct binding of WD40 repeat of Embryonic Ectoderm Development (EED) in the Polycomb Repressor Complex 2 (PRC2) to the histone H3 tail regulates the H3K27 methyltrasnferase activity of PRC2. The binding activity is required for the methylation of H3K27 over long distance of a Drosophila chromosome from the UBX (Hox gene) promoter to the PBX upstream enhancer region, implying EED initiates propagation of this repressive mark. PMID- 20008335 TI - NANOS: a germline stem cell's Guardian Angel. AB - NANOS is known to be required for germline cell development in a variety of animal species and for the maintenance of germline stem cells in Drosophila. The recent study by Sada et al. has demonstrated that NANOS2, one of the three mammalian homologues, is required intrinsically for maintaining adult mouse spermatogonial stem cell self-renewal. PMID- 20008336 TI - The dazzle in germ cell differentiation. AB - Embryonic stem cells have demonstrated the capacity to differentiate into germ cells in vitro. Until recently, the molecular basis of early post-meiotic germ cell development was largely unknown. Now, two reports including one published here recently, have demonstrated the significant contribution of Dazl in the differentiation of embryonic stem cells into pre- and post-meiotic germ cells. Although factors that Dazl influences during this process have been identified, the underlying mechanisms warrant future studies. PMID- 20008337 TI - Coronary artery fistula as major source of right lung circulation in a patient with isolated right pulmonary artery agenesis. PMID- 20008338 TI - Cardiomyocyte disintegration during Anderson-Fabry's disease. PMID- 20008339 TI - Analysis of galactosemia-linked mutations of GALT enzyme using a computational biology approach. AB - We describe the prediction of the structural and functional effects of mutations on the enzyme galactose-1-phosphate uridyltransferase related to the genetic disease galactosemia, using a fully computational approach. One hundred and seven single-point mutants were simulated starting from the structural model of the enzyme obtained by homology modeling methods. Several bioinformatics programs were then applied to each resulting mutant protein to analyze the effect of the mutations. The mutations have a direct effect on the active site, or on the dimer assembly and stability, or on the monomer stability. We describe how mutations may exert their effect at a molecular level by altering H-bonds, salt bridges, secondary structure or surface features. The alteration of protein stability, at level of monomer and/or dimer, is the main effect observed. We found an agreement between our results and the functional experimental data available in literature for some mutants. The data and analyses for all the mutants are fully available in the web-accessible database hosted at http://bioinformatica.isa.cnr.it/GALT. PMID- 20008340 TI - Was Charles Darwin interested in the brain? PMID- 20008341 TI - Does Epstein-Barr virus infection in the brain drive the development of multiple sclerosis? PMID- 20008342 TI - Continued diagnosis of idiopathic haemorrhagic diathesis in calves. PMID- 20008343 TI - Lameness and effusion of the tarsocrural joints after arthroscopy of osteochondritis dissecans in horses. AB - The presence and degree of tarsocrural joint effusion and lameness, and the result of a hindlimb flexion test were scored subjectively in 79 horses before, and six weeks to 20 months after, arthroscopic surgery to remove osteochondritis dissecans fragments. The scores of all three variables improved significantly after surgery. The score reductions for the right and left hindlimbs, respectively, were 82 per cent and 95 per cent for lameness, 48 per cent and 41 per cent for joint effusion, and 89 per cent and 84 per cent for reaction to flexion (P < or = 0.01). The oldest horses reacted more favourably to the operation, as measured by the reaction to the flexion test, but age was not significantly related to changes in lameness or joint effusion. There was no significant correlation between the time of follow-up examination and the effect of surgery on lameness and reaction to flexion, but an increased time to follow up was associated with decreased joint effusion. PMID- 20008344 TI - Effects of a deoxynivalenol-contaminated diet on the reproductive performance and immunoglobulin concentrations in pigs. AB - Two groups of 10 pregnant gilts (89 +/- 2 days gestation) were fed either an experimental diet that contained 5.08 mg/kg deoxynivalenol, 0.09 mg/kg zearalenone and 21.6 mg/kg fusaric acid, or a control diet that contained 0.29 mg/kg deoxynivalenol, <0.02 mg/kg zearalenone and <0.77 mg/kg fusaric acid. The concentrations of immunoglobulins were measured in sera of the gilts and in the colostrum and serum of the piglets by radial immunodiffusion. The feed consumption of the sows fed the experimental diet was significantly lower and the overall growth rate of their piglets was significantly reduced. On average, parturition took 80 minutes longer in sows fed the experimental diet. On day 17 after parturition, the concentration of IgM in the serum of the experimental gilts was significantly higher, but the concentration of IgA in their colostrum was significantly lower, than in the control gilts. In the serum of the piglets 12, 24 and 48 hours after first suckling, the concentrations of IgA and IgG were significantly lower in those farrowed by the sows fed the experimental diet than in those farrowed by the sows fed the control diet. PMID- 20008345 TI - Four cases of omental herniation in cattle. AB - Four cases in cattle of omental herniation through an acquired omental rent are described. Clinical signs were indistinguishable from other causes of mechanical ileus and exploratory laparotomy was necessary to establish a diagnosis. In all cases a variably sized portion of jejunal loops was obstructed in an omental rent in the deep layer of the greater omentum. In two cases simple reduction was possible, and in one case incision of the hernial ring was necessary before reduction could be performed. Resection was necessary only in a calf, in which the incarceration was complicated by severe abomasal distension and local peritonitis. In all four cases the omental rent was closed by a serosa-serosa suture. Three cases made an uneventful recovery and returned to normal production, but one of these animals died three months postoperatively from an unknown cause. The calf was euthanased two days postoperatively because of persistent ileus. PMID- 20008346 TI - Effect of levamisole on the humoral immune response against rabies in cattle. PMID- 20008347 TI - Use of histamine phosphate for the interpretation of intradermal skin tests in dogs. PMID- 20008348 TI - Pets and carbon footprints. PMID- 20008349 TI - Alternative business models for veterinary practice. PMID- 20008351 TI - Clinical governance. PMID- 20008352 TI - Call for national action to control bovine viral diarrhoea. PMID- 20008353 TI - Molecular details of Bax activation, oligomerization, and membrane insertion. AB - Bax and Bid are pro-apoptotic members of the Bcl-2 protein family. Upon cleavage by caspase-8, Bid activates Bax. Activated Bax inserts into the mitochondrial outer membrane forming oligomers which lead to membrane poration, release of cytochrome c, and apoptosis. The detailed mechanism of Bax activation and the topology and composition of the oligomers are still under debate. Here molecular details of Bax activation and oligomerization were obtained by application of several biophysical techniques, including atomic force microscopy, cryoelectron microscopy, and particularly electron paramagnetic resonance (EPR) spectroscopy performed on spin-labeled Bax. Incubation with detergents, reconstitution, and Bid-triggered insertion into liposomes were found to be effective in inducing Bax oligomerization. Bid was shown to activate Bax independently of the stoichiometric ratio, suggesting that Bid has a catalytic function and that the interaction with Bax is transient. The formation of a stable dimerization interface involving two Bcl-2 homology 3 (BH3) domains was found to be the nucleation event for Bax homo-oligomerization. Based on intermolecular distance determined by EPR, a model of six adjacent Bax molecules in the oligomer is presented where the hydrophobic hairpins (helices alpha5 and alpha6) are equally spaced in the membrane and the two BH3 domains are in close vicinity in the dimer interface, separated by >5 nm from the next BH3 pairs. PMID- 20008354 TI - Bill Harvey retires. PMID- 20008356 TI - Epigenetics and transgenerational transfer: a physiological perspective. AB - Epigenetics, the transgenerational transfer of phenotypic characters without modification of gene sequence, is a burgeoning area of study in many disciplines of biology. However, the potential impact of this phenomenon on the physiology of animals is not yet broadly appreciated, in part because the phenomenon of epigenetics is not typically part of the design of physiological investigations. Still enigmatic and somewhat ill defined is the relationship between the overarching concept of epigenetics and interesting transgenerational phenomena (e.g. 'maternal/parental effects') that alter the physiological phenotype of subsequent generations. The lingering effect on subsequent generations of an initial environmental disturbance in parent animals can be profound, with genes continuing to be variously silenced or expressed without an associated change in gene sequence for many generations. Known epigenetic mechanisms involved in this phenomenon include chromatin remodeling (DNA methylation and histone modification), RNA-mediated modifications (non-coding RNA and microRNA), as well as other less well studied mechanisms such as self-sustaining loops and structural inheritance. In this review we: (1) discuss how the concepts of epigenetics and maternal effects both overlap with, and are distinct from, each other; (2) analyze examples of existing animal physiological studies based on these concepts; and (3) offer a construct by which to integrate these concepts into the design of future investigations in animal physiology. PMID- 20008357 TI - Regulation of sarcoendoplasmic reticulum Ca2+-ATPase (SERCA) in turtle muscle and liver during acute exposure to anoxia. AB - The freshwater turtle Trachemys scripta elegans naturally tolerates extended periods of anoxia during winter hibernation at the bottom of ice-locked ponds. Survival in this anoxic state is facilitated by a profound depression of metabolic rate. As calcium levels are known to be elevated in anoxic turtles, and ion pumping is an ATP-expensive process, we proposed that activity of the sarcoendoplasmic reticulum Ca(2+)-ATPase (SERCA) would be reduced in muscle and liver of T. s. elegans during acute (up to 20 h) exposure to anoxia. SERCA activity decreased approximately 30% in liver and approximately 40% in muscle after 1 h anoxia exposure and was approximately 50% lower after 20 h of anoxia exposure in both tissues, even though SERCA protein levels did not change. SERCA kinetic parameters (increased substrate K(m) values, increased Arrhenius activation energy) were indicative of a less active enzyme form under anoxic conditions. Interestingly, the less active SERCA in anoxic turtles featured greater stability than the enzyme from normoxic animals as determined by both kinetic analysis (effect of low pH and low temperatures on K(m) MgATP) and conformational resistance to urea denaturation. The quick time course of deactivation and the stable changes in kinetic parameters that resulted suggested that SERCA was regulated by a post-translational mechanism. In vitro experiments indicated that SERCA activity could be blunted by protein phosphorylation and enhanced by dephosphorylation in a tissue-specific manner. PMID- 20008358 TI - Individual variation and repeatability in aerobic and anaerobic swimming performance of European sea bass, Dicentrarchus labrax. AB - Studies of inter-individual variation in fish swimming performance may provide insight into how selection has influenced diversity in phenotypic traits. We investigated individual variation and short-term repeatability of individual swimming performance by wild European sea bass in a constant acceleration test (CAT). Fish were challenged with four consecutive CATs with 5 min rest between trials. We measured maximum anaerobic speed at exhaustion (U(CAT)), gait transition speed from steady aerobic to unsteady anaerobic swimming (U(gt)), routine metabolic rate (RMR), post-CAT maximum metabolic rate (MMR), aerobic scope and recovery time from the CATs. Fish achieved significantly higher speeds during the first CAT (U(CAT)=170 cm s(-1)), and had much more inter-individual variation in performance (coefficient of variation, CV=18.43%) than in the subsequent three tests (U(CAT)=134 cm s(-1); CV=7.3%), which were very repeatable among individuals. The individual variation in U(CAT) in the first trial could be accounted for almost exclusively by variation in anaerobic burst-and-coast performance beyond U(gt). The U(gt) itself varied substantially between individuals (CV=11.4%), but was significantly repeatable across all four trials. Individual RMR and MMR varied considerably, but the rank order of post-CAT MMR was highly repeatable. Recovery rate from the four CATs was highly variable and correlated positively with the first U(CAT) (longer recovery for higher speeds) but negatively with RMR and aerobic scope (shorter recovery for higher RMR and aerobic scope). This large variation in individual performance coupled with the strong correlations between some of the studied variables may reflect divergent selection favouring alternative strategies for foraging and avoiding predation. PMID- 20008359 TI - Ecophysiological response of Adelie penguins facing an experimental increase in breeding constraints. AB - Foraging strategies play a key role in breeding effort. Little is known, however, about their connection with hormonal and nutritional states, especially when breeding constraints vary. Here, we experimentally increased foraging costs and thus breeding constraints by handicapping Adelie penguins (Pygoscelis adeliae) with dummy devices representing 3-4% of the penguins' cross-sectional area. We examined food-related stress (via plasma corticosterone concentration) and nutritional state (via metabolite levels). Concurrently, we investigated the use of ecological niches via the isotopic signature of red blood cells indicating the trophic position (delta(15)N) and the spatial distribution (delta(13)C) of penguins. Handicapped birds performed approximately 70% longer foraging trips and lost approximately 60% more body mass than controls and their partners. However, corticosterone levels and the nutritional state were unchanged. The isotopic signature revealed that males and females differed in their foraging behaviour: upper trophic levels contributed more in the males' diet, who foraged in more pelagic areas. Handicapped and partner birds adopted the same strategy at sea: a shift towards higher delta(13)C values suggested that they foraged in more coastal areas than controls. This change in foraging decisions may optimize feeding time by decreasing travelling time. This may partly compensate for the presumed lower foraging efficiency of handicapped birds and for the energetic debt of their partners who had to fast approximately 70% longer on the nest. We propose that this flexible use of ecological niches may allow birds facing increased breeding constraints to avoid chronic stress and to minimize the impact on their body condition. PMID- 20008360 TI - The distinction between retractor and protractor muscles of the freshwater snail's male organ has no physiological basis. AB - Many animals are equipped with organs that can be everted, a notable example being male copulatory organs. The ability to protrude or evert an organ generally requires protractor and retractor muscles. Male copulatory behaviour of the pond snail Lymnaea stagnalis (L.) involves eversion (protraction) and retraction of the relatively large penis-carrying organ. For this preputium, protractor and retractor muscle bands have been defined, which implies eversion and retraction through the activity of these muscle bands. However, no physiological data are available that confirm that the terms protractor and retractor are appropriate. To test whether eversion and retraction are possible without protractor and/or retractor muscle bands, lesion experiments were performed. The results show that with either one or several muscle bands lesioned, snails were still capable of everting their preputium and using it for copulation. However, the majority of animals that had six or more muscle bands lesioned were unable to retract its preputium. Hence, retractor muscle bands serve their designated function whereas protractor muscle bands do not. We therefore suggest that a different terminology is used in which all muscle bands are retractors and, based on their location, are either called distal or proximal retractors. The findings furthermore indicate that the preputium muscle bands are normally contracted, possibly in a catch state, retaining the organ inside without high-energy expenditure. PMID- 20008361 TI - Slight differences in incubation temperature affect early growth and stress endocrinology of wood duck (Aix sponsa) ducklings. AB - Early developmental experiences, such as incubation conditions, can have important consequences for post-hatching fitness in birds. Although the effects of incubation temperature on phenotype of avian hatchlings are poorly understood, recent research suggests that subtle changes in incubation conditions can influence hatchling characteristics, including body size and condition. We designed an experiment to explore the effects of incubation temperature on hatching success, survival to 9 days post hatch, growth and the hypothalamo pituitary-adrenal (HPA) axis in wood ducks (Aix sponsa). Wood duck eggs were collected from nest boxes and experimentally incubated at three temperatures (35.0, 35.9 and 37.0 degrees C), each falling within the range of temperatures of naturally incubated wood duck nests. Survival and growth were monitored in ducklings fed ad libitum for 9 days post hatch. In addition, baseline and stress induced plasma corticosterone concentrations were measured in 2 and 9 day old ducklings. Hatching success and survival to 9 days was greatest in ducks incubated at the intermediate temperature. Ducklings incubated at 35.9 degrees C and 37.0 degrees C had 43% higher growth rates than ducklings incubated at 35.0 degrees C. In addition, ducklings incubated at 35.0 degrees C had higher baseline (17-50%) and stress-induced (32-84%) corticosterone concentrations than ducklings incubated at 35.9 degrees C and 37.0 degrees C at 2 and 9 days post hatch. We also found a significant negative correlation between body size and plasma corticosterone concentrations (baseline and stress-induced) in 9 day old ducklings. To our knowledge, this is the first study to demonstrate that thermal conditions experienced during embryonic development can influence the HPA axis of young birds. Our results illustrate that subtle changes (<1.0 degrees C) in the incubation environment can have important consequences for physiological traits important to fitness. PMID- 20008362 TI - Investigation of the potential for vascular bubble formation in a repetitively diving dolphin. AB - The production of venous gas emboli (VGE) resulting from altered dive behavior is postulated as contributing to the stranding of beaked whales exposed to mid frequency active sonar. To test whether nitrogen gas uptake during repetitive breath-hold diving is sufficient for asymptomatic VGE formation in odontocetes, a bottlenose dolphin (Tursiops truncatus Montagu) was trained to perform 10-12 serial dives with 60 s surface intervals to depths of 30, 50, 70 or 100 m. The dolphin remained at the bottom depth for 90 s on each dive. Doppler and/or two dimensional imaging ultrasound did not detect VGE in the portal and brachiocephalic veins following a dive series. Van Slyke analyses of serial, post dive blood samples drawn from the fluke yielded blood nitrogen partial pressure (P(N(2))) values that were negligibly different from control samples. Mean heart rate (HR; +/-1 s.d.) recorded during diving was 50+/-3 beats min(-1) and was not significantly different between the 50, 70 and 100 m dive sessions. The absence of VGE and elevated blood P(N(2)) during post-dive periods do not support the hypothesis that N(2) supersaturation during repetitive dives contributes to VGE formation in the dolphin. The diving HR pattern and the presumed rapid N(2) washout during the surface-interval tachycardia probably minimized N(2) accumulation in the blood during dive sessions. PMID- 20008363 TI - In-line motion causes high thrust and efficiency in flapping foils that use power downstroke. AB - We show experimentally that flapping foil kinematics consisting of a power downstroke and a feathering upstroke together with a properly timed in-line motion, similar to those employed in forelimb propulsion of sea turtles, can produce high thrust and be hydrodynamically as efficient as symmetrically flapping foils. The crucial parameter for such asymmetrically flapping foils is a properly sized and timed in-line motion, whose effect is quantified by a new parameter, the advance angle, defined as the angle of the foil trajectory with respect to the horizontal, evaluated at the middle of the power downstroke. We show, in particular, that optimal efficiency in high aspect ratio rigid foils, accompanied by significant thrust production, is obtained for Strouhal numbers in the range 0.2-0.6 for Reynolds number equal to 13,000, and for values of the advance angle around 0.55pi (100 deg.). The optimized kinematics consist of the foil moving back axially during the downstroke, in the direction of the oncoming flow, and rotating with a large pitch angle. This causes the force vector to rotate and become nearly parallel to the steady flow, thus providing a large thrust and a smaller transverse force. During the upstroke, the foil is feathering while it moves axially forward, i.e. away from the vorticity shed during the power stroke; as a result, the transverse force remains relatively small and no large drag force is produced. Observations from turtles confirm qualitatively the findings from the foil experiments. PMID- 20008364 TI - First evidence of fine colour discrimination ability in ants (Hymenoptera, Formicidae). AB - In the present study, we report the first evidence that ants discriminate and learn perceptually close colour stimuli. Foragers of the ant species Cataglyphis aenescens and Formica cunicularia were trained in a Y-maze choice apparatus to monochromatic light stimuli of a constant intensity associated with a food reward. Two stimuli, with a mean wavelength of 40 nm perceptual distance, were chosen from the UV (340 nm vs 380 nm) and the green (510 nm vs 550 nm) range because these species are UV-green dichromats. Foragers were trained with two conditioning paradigms [absolute conditioning (AC) and differential conditioning (DC)]. In the UV range, C. aenescens foragers failed to discriminate when presented with a small colour difference in both training procedures. Foragers also failed in the green range when trained with AC but showed significant bias towards the rewarded stimulus when trained with DC. Formica cunicularia foragers achieved the task in the UV range when trained with DC only. In the green range, F. cunicularia foragers showed clear preference for the rewarded stimulus in both training conditioning procedures. Foragers never failed in choosing the rewarded stimulus in DC even when the intensity of the rewarded stimulus was reduced by one log unit. This clearly indicates that DC is of paramount importance to discriminate perceptually close colour stimuli. PMID- 20008365 TI - Prioritizing blood flow: cardiovascular performance in response to the competing demands of locomotion and digestion for the Burmese python, Python molurus. AB - Individually, the metabolic demands of digestion or movement can be fully supported by elevations in cardiovascular performance, but when occurring simultaneously, vascular perfusion may have to be prioritized to either the gut or skeletal muscles. Burmese pythons (Python molurus) experience similar increases in metabolic rate during the digestion of a meal as they do while crawling, hence each would have an equal demand for vascular supply when these two actions are combined. To determine, for the Burmese python, whether blood flow is prioritized when snakes are digesting and moving, we examined changes in cardiac performance and blood flow in response to digestion, movement, and the combination of digestion and movement. We used perivascular blood flow probes to measure blood flow through the left carotid artery, dorsal aorta, superior mesenteric artery and hepatic portal vein, and to calculate cardiac output, heart rate and stroke volume. Fasted pythons while crawling experienced a 2.7- and 3.3 fold increase, respectively, in heart rate and cardiac output, and a 66% decrease in superior mesenteric flow. During the digestion of a rodent meal equaling in mass to 24.7% of the snake's body mass, heart rate and cardiac output increased by 3.3- and 4.4-fold, respectively. Digestion also resulted in respective 11.6- and 14.1-fold increases in superior mesenteric and hepatic portal flow. When crawling while digesting, cardiac output and dorsal aorta flow increased by only 21% and 9%, respectively, a modest increase compared with that when they start to crawl on an empty stomach. Crawling did triggered a significant reduction in blood flow to the digesting gut, decreasing superior mesenteric and hepatic portal flow by 81% and 47%, respectively. When faced with the dual demands of digestion and crawling, Burmese pythons prioritize blood flow, apparently diverting visceral supply to the axial muscles. PMID- 20008366 TI - On the role of form and kinematics on the hydrodynamics of self-propelled body/caudal fin swimming. AB - We carry out fluid-structure interaction simulations of self-propelled virtual swimmers to investigate the effects of body shape (form) and kinematics on the hydrodynamics of undulatory swimming. To separate the effects of form and kinematics, we employ four different virtual swimmers: a carangiform swimmer (i.e. a mackerel swimming like mackerel do in nature); an anguilliform swimmer (i.e. a lamprey swimming like lampreys do in nature); a hybrid swimmer with anguilliform kinematics but carangiform body shape (a mackerel swimming like a lamprey); and another hybrid swimmer with carangiform kinematics but anguilliform body shape (a lamprey swimming like a mackerel). By comparing the performance of swimmers with different kinematics but similar body shapes we study the effects of kinematics whereas by comparing swimmers with similar kinematics but different body shapes we study the effects of form. We show that the anguilliform kinematics not only reaches higher velocities but is also more efficient in the viscous (Re approximately 10(2)) and transitional (Re approximately 10(3)) regimes. However, in the inertial regime (Re=infinity) carangiform kinematics achieves higher velocities and is also more efficient than the anguilliform kinematics. The mackerel body achieves higher swimming speeds in all cases but is more efficient in the inertial regime only whereas the lamprey body is more efficient in the transitional regime. We also show that form and kinematics have little overall effect on the 3-D structure of the wake (i.e. single vs double row vortex streets), which mainly depends on the Strouhal number. Nevertheless, body shape is found to somewhat affect the small-scale features and complexity of the vortex rings shed by the various swimmers. PMID- 20008367 TI - Lateral line diversity among ecologically divergent threespine stickleback populations. AB - The lateral line is a mechanoreceptive sensory system that allows fish to sense objects and motion in their local environment. Variation in lateral line morphology may allow fish in different habitats to differentially sense and respond to salient cues. Threespine sticklebacks (Gasterosteus aculeatus) occupy a diverse range of aquatic habitats; we therefore hypothesized that populations within the G. aculeatus species complex might show variation in the morphology of the lateral line sensory system. We sampled 16 threespine stickleback populations from marine, stream and lake (including benthic and limnetic) habitats and examined the distribution, type and number of neuromasts on different regions of the body. We found that the threespine stickleback has a reduced lateral line canal system, completely lacking canal neuromasts. Although the arrangement of lines of superficial neuromasts on the body was largely the same in all populations, the number of neuromasts within these lines varied across individuals, populations and habitats. In pairwise comparisons between threespine sticklebacks adapted to divergent habitats, we found significant differences in neuromast number. Stream residents had more neuromasts than marine sticklebacks living downstream in the same watershed. In two independent lakes, benthic sticklebacks had more trunk neuromasts than sympatric limnetic sticklebacks, providing evidence for parallel evolution of the lateral line system. Our data provide the first demonstration that the lateral line sensory system can vary significantly between individuals and among populations within a single species, and suggest that this sensory system may experience different selection regimes in alternative habitats. PMID- 20008369 TI - Variation of the neurofilament medium KSP repeat sub-domain across mammalian species: implications for altering axonal structure. AB - The evolution of larger mammals resulted in a corresponding increase in peripheral nerve length. To ensure optimal nervous system functionality and survival, nerve conduction velocities were likely to have increased to maintain the rate of signal propagation. Increases of conduction velocities may have required alterations in one of the two predominant properties that affect the speed of neuronal transmission: myelination or axonal diameter. A plausible mechanism to explain faster conduction velocities was a concomitant increase in axonal diameter with evolving axonal length. The carboxy terminal tail domain of the neurofilament medium subunit is a determinant of axonal diameter in large caliber myelinated axons. Sequence analysis of mammalian orthologs indicates that the neurofilament medium carboxy terminal tail contains a variable lysine-serine proline (KSP) repeat sub-domain flanked by two highly conserved sub-domains. The number of KSP repeats within this region of neurofilament medium varies among species. Interestingly, the number of repeats does not change within a species, suggesting that selective pressure conserved the number of repeats within a species. Mapping KSP repeat numbers onto consensus phylogenetic trees reveals independent KSP expansion events across several mammalian clades. Linear regression analyses identified three subsets of mammals, one of which shows a positive correlation in the number of repeats with head-body length. For this subset of mammals, we hypothesize that variations in the number of KSP repeats within neurofilament medium carboxy terminal tail may have contributed to an increase in axonal caliber, increasing nerve conduction velocity as larger mammals evolved. PMID- 20008368 TI - Identification of a calcitonin-like diuretic hormone that functions as an intrinsic modulator of the American lobster, Homarus americanus, cardiac neuromuscular system. AB - In insects, a family of peptides with sequence homology to the vertebrate calcitonins has been implicated in the control of diuresis, a process that includes mixing of the hemolymph. Here, we show that a member of the insect calcitonin-like diuretic hormone (CLDH) family is present in the American lobster, Homarus americanus, serving, at least in part, as a powerful modulator of cardiac output. Specifically, during an ongoing EST project, a transcript encoding a putative H. americanus CLDH precursor was identified; a full-length cDNA was subsequently cloned. In silico analyses of the deduced prepro-hormone predicted the mature structure of the encoded CLDH to be GLDLGLGRGFSGSQAAKHLMGLAAANFAGGPamide (Homam-CLDH), which is identical to a known Tribolium castaneum peptide. RT-PCR tissue profiling suggests that Homam-CLDH is broadly distributed within the lobster nervous system, including the cardiac ganglion (CG), which controls the movement of the neurogenic heart. RT-PCR analysis conducted on pacemaker neuron- and motor neuron-specific cDNAs suggests that the motor neurons are the source of the CLDH message in the CG. Perfusion of Homam-CLDH through the isolated lobster heart produced dose-dependent increases in both contraction frequency and amplitude and a dose-dependent decrease in contraction duration, with threshold concentrations for all parameters in the range 10(-11) to 10(-10) mol l(-1) or less, among the lowest for any peptide on this system. This report is the first documentation of a decapod CLDH, the first demonstration of CLDH bioactivity outside the Insecta, and the first detection of an intrinsic neuropeptide transcript in the crustacean CG. PMID- 20008370 TI - Fibre type-specific expression patterns of myosin heavy chain genes in adult torafugu Takifugu rubripes muscles. AB - Comprehensive in silico studies, based on the total fugu genome database, which was the first to appear in fish, revealed that torafugu Takifugu rubripes contains 20 sarcomeric myosin heavy chain (MYH) genes (MYH genes) (Ikeda et al., 2007). The present study was undertaken to identify MYH genes that would be expressed in adult muscles. In total, seven MYH genes were found by screening cDNA clone libraries constructed from fast, slow and cardiac muscles. Three MYH genes, fast-type MYH(M86-1), slow-type MYH(M8248) and slow/cardiac-type MYH(M880), were cloned exclusively from fast, slow and cardiac muscles, respectively. Northern blot hybridization substantiated their specific expression, with the exception of MYH(M880). In contrast, transcripts of fast type MYH(M2528-1) and MYH(M1034) were found in both fast and slow muscles as revealed by cDNA clone library and northern blot techniques. This result was supported by in situ hybridization analysis using specific RNA probes, where transcripts of fast-type MYH(M2528-1) were expressed in fast fibres with small diameters as well as in fibres of superficial slow muscle with large diameters adjacent to fast muscle. Transcripts of fast-type MYH(M86-1) were expressed in all fast fibres with different diameters, whereas transcripts of slow-type MYH(M8248) were restricted to fibres with small diameters located in a superficial part of slow muscle. Interestingly, histochemical analyses showed that fast fibres with small diameters and slow fibres with large diameters both contained acid-stable myofibrillar ATPase, suggesting that these fibres have similar functions, possibly in the generation of muscle fibres irrespective of their fibre types. PMID- 20008371 TI - Scanning sonar of rolling porpoises during prey capture dives. AB - Dolphins and porpoises have excellent biosonar ability, which they use for navigation, ranging and foraging. However, the role of biosonar in free-ranging small cetaceans has not been fully investigated. The biosonar behaviour and body movements of 15 free-ranging finless porpoises (Neophocaena phocaenoides) were observed using electronic tags attached to the animals. The porpoises often rotated their bodies more than 60 deg., on average, around the body axis in a dive bout. This behaviour occupied 31% of the dive duration during 186 h of effective observation time. Rolling dives were associated with extensive searching effort, and 23% of the rolling dive time was phonated, almost twice the phonation ratio of upright dives. Porpoises used short inter-click interval sonar 4.3 times more frequently during rolling dives than during upright dives. Sudden speed drops, which indicated that an individual turned around, occurred 4.5 times more frequently during rolling dives than during upright dives. Together, these data suggest that the porpoises searched extensively for targets and rolled their bodies to enlarge the search area by changing the narrow beam axis of the biosonar. Once a possible target was detected, porpoises frequently produced short-range sonar sounds. Continuous searching for prey and frequent capture trials appeared to occur during rolling dives of finless porpoises. In contrast, head movements ranging +/-2 cm, which can also change the beam axis, were regularly observed during both dives. Head movements might assist in instant assessment of the arbitrary direction by changing the beam axis rather than prey searching and pursuit. PMID- 20008372 TI - Heart rate as a predictor of energy expenditure in undisturbed fasting and incubating penguins. AB - Heart rate (f(H)) measurement offers the possibility to monitor energy expenditure (EE) in wild animals if the EE/f(H) relationship for the species, physiological stages and activities of interest is known. This relationship has been extensively studied using oxygen consumption rate ( ) measurement in captive, repeatedly handled king penguins (Aptenodytes patagonicus). Unfortunately, the potential effects of stress on the observed relationships resulting from handling and confinement were not considered. This study is the first involving undisturbed animals, and determines the EE/f(H) relationship in naturally fasting and freely incubating or captivity-acclimatized male and female king penguins. EE determination was based on (1) the measurement of body mass loss during periods of phase II fasting, and (2) the calculation of its energy equivalent from changes in body composition, i.e. 23.9 kJ g(-1). f(H) levels in freely incubating and captivity-acclimatized birds were found to be 50-70% lower than those previously reported for resting king penguins during measurements. Significant EE/f(H) relationships were found in freely incubating and captive males and females (R(2)=0.59 to 0.84), with no difference observed between genders. The best overall relationship was obtained by including fasting duration (t, days) in the model: EE=818+43.7xf(H)+36.3t-1.4txf(H) (R(2)=0.91). This equation yielded EE estimates approximately 26% higher than the previously reported 'best' predictive equation in king penguins, and even more so when f(H) was low. This result suggests that stress induces a disproportionate increase of f(H) vs O(2) consumption, and that the use of EE/f(H) relationships obtained in stressed birds could lead to underestimated EE values. PMID- 20008373 TI - Gravikinesis in Stylonychia mytilus is based on membrane potential changes. AB - The graviperception of the hypotrichous ciliate Stylonychia mytilus was investigated using electrophysiological methods and behavioural analysis. It is shown that Stylonychia can sense gravity and thereby compensates sedimentation rate by a negative gravikinesis. The graviresponse consists of a velocity regulating physiological component (negative gravikinesis) and an additional orientational component. The latter is largely based on a physical mechanism but might, in addition, be affected by the frequency of ciliary reversals, which is under physiological control. We show that the external stimulus of gravity is transformed to a physiological signal, activating mechanosensitive calcium and potassium channels. Earlier electrophysiological experiments revealed that these ion channels are distributed in the manner of two opposing gradients over the surface membrane. Here, we show, for the first time, records of gravireceptor potentials in Stylonychia that are presumably based on this two-gradient system of ion channels. The gravireceptor potentials had maximum amplitudes of approximately 4 mV and slow activation characteristics (0.03 mV s(-1)). The presumptive number of involved graviperceptive ion channels was calculated and correlates with the analysis of the locomotive behaviour. PMID- 20008374 TI - Molt cycle regulation of protein synthesis in skeletal muscle of the blackback land crab, Gecarcinus lateralis, and the differential expression of a myostatin like factor during atrophy induced by molting or unweighting. AB - In decapod crustaceans, claw muscle undergoes atrophy in response to elevated ecdysteroids while thoracic muscle undergoes atrophy in response to unweighting. The signaling pathways that regulate muscle atrophy in crustaceans are largely unknown. Myostatin is a negative regulator of muscle growth in mammals, and a myostatin-like cDNA is preferentially expressed in muscle of the land crab, Gecarcinus lateralis (Gl-Mstn). Contrary to prediction, levels of Gl-Mstn mRNA decreased dramatically in both the claw closer and weighted thoracic muscles when molting was induced by either eyestalk ablation (ESA) or multiple limb autotomy (MLA). However, the effect of molt induction was greater in the claw muscle. By late premolt, Gl-Mstn mRNA in the claw muscle decreased 81% and 94% in ESA and MLA animals, respectively, and was negatively correlated with ecdysteroids. Gl Mstn mRNA in thoracic muscle decreased 68% and 82% in ESA and MLA animals, respectively, but was only weakly correlated with ecdysteroid. Claw and thoracic muscles also differed to varying extents in the expression of ecdysteroid receptor (Gl-EcR and Gl-RXR), elongation factor-2 (Gl-EF-2), and calpain T (Gl CalpT) in response to molt induction, but levels of the four transcripts were not correlated with ecdysteroid. The downregulation of Gl-Mstn expression in premolt claw muscle coincided with 11- and 13-fold increases in protein synthesis in the myofibrillar and soluble protein fractions, respectively. Furthermore, the rate of the increase in the synthesis of soluble proteins was greater than that of myofibrillar proteins during early premolt (1.4:1, soluble:myofibrillar), but the two were equivalent during late premolt. By contrast, Gl-Mstn mRNA increased 3 fold and Gl-CalpT mRNA decreased 40% in unweighted thoracic muscle; there was little or no effect on Gl-EF-2, Gl-EcR, and Gl-RXR mRNA levels. These data indicate that Gl-Mstn expression is negatively regulated by both ecdysteroids and load-bearing contractile activity. The downregulation of Gl-Mstn in claw muscle may induce the elevated protein turnover associated with remodeling of the contractile apparatus during molt-induced atrophy. The upregulation of Gl-Mstn in unweighted thoracic muscle suggests that this factor is also involved in disuse atrophy when hemolymph ecdysteroid levels are low. PMID- 20008375 TI - Atypical neural self-representation in autism. AB - The 'self' is a complex multidimensional construct deeply embedded and in many ways defined by our relations with the social world. Individuals with autism are impaired in both self-referential and other-referential social cognitive processing. Atypical neural representation of the self may be a key to understanding the nature of such impairments. Using functional magnetic resonance imaging we scanned adult males with an autism spectrum condition and age and IQ matched neurotypical males while they made reflective mentalizing or physical judgements about themselves or the British Queen. Neurotypical individuals preferentially recruit the middle cingulate cortex and ventromedial prefrontal cortex in response to self compared with other-referential processing. In autism, ventromedial prefrontal cortex responded equally to self and other, while middle cingulate cortex responded more to other-mentalizing than self-mentalizing. These atypical responses occur only in areas where self-information is preferentially processed and does not affect areas that preferentially respond to other referential information. In autism, atypical neural self-representation was also apparent via reduced functional connectivity between ventromedial prefrontal cortex and areas associated with lower level embodied representations, such as ventral premotor and somatosensory cortex. Furthermore, the magnitude of neural self-other distinction in ventromedial prefrontal cortex was strongly related to the magnitude of early childhood social impairments in autism. Individuals whose ventromedial prefrontal cortex made the largest distinction between mentalizing about self and other were least socially impaired in early childhood, while those whose ventromedial prefrontal cortex made little to no distinction between mentalizing about self and other were the most socially impaired in early childhood. These observations reveal that the atypical organization of neural circuitry preferentially coding for self-information is a key mechanism at the heart of both self-referential and social impairments in autism. PMID- 20008376 TI - Effect of listening to Nellie the Elephant during CPR training on performance of chest compressions by lay people: randomised crossover trial. AB - OBJECTIVES: To determine whether listening to music during cardiopulmonary resuscitation (CPR) training increases the proportion of lay people delivering chest compressions of 100 per minute. DESIGN: Prospective randomised crossover trial. SETTING: Large UK university. PARTICIPANTS: 130 volunteers (81 men) recruited on an opportunistic basis. Exclusion criteria included age under 18, trained health professionals, and cardiopulmonary resuscitation (CPR) training within the past three months. INTERVENTIONS: Volunteers performed three sequences of one minute of continuous chest compressions on a skill meter resuscitation manikin accompanied by no music, repeated choruses of Nellie the Elephant (Nellie), and That's the Way (I like it) (TTW) according to a pre-randomised order. MAIN OUTCOME MEASURES: Rate of chest compressions delivered (primary outcome), depth of compressions, proportion of incorrect compressions, and type of error. RESULTS: Median (interquartile range) compression rates were 110 (93 119) with no music, 105 (98-107) with Nellie, and 109 (103-110) with TTW. There were significant differences within groups between Nellie v no music and Nellie v TTW (P<0.001) but not no music v TTW (P=0.055). A compression rate of between 95 and 105 was achieved with no music, Nellie, and TTW for 15/130 (12%), 42/130 (32%), and 12/130 (9%) attempts, respectively. Differences in proportions were significant for Nellie v no music and Nellie v TTW (P<0.001) but not for no music v TTW (P=0.55). Relative risk for a compression rate between 95 and 105 was 2.8 (95% confidence interval 1.66 to 4.80) for Nellie v no music, 0.8 (0.40 to 1.62) for TTW v no music, and 3.5 (1.97 to 6.33) for Nellie v TTW. The number needed to treat for listening to Nellie v no music was 5 (4 to 10)-that is, the number of cardiac arrests required during which lay responders listen to Nellie to facilitate one patient receiving compressions at the correct rate (v no music) would be between four and 10. A greater proportion of compressions were too shallow when participants listened to Nellie v no music (56% v 47%, P=0.022). CONCLUSIONS: Listening to Nellie the Elephant significantly increased the proportion of lay people delivering compression rates at close to 100 per minute. Unfortunately it also increased the proportion of compressions delivered at an inadequate depth. As current resuscitation guidelines give equal emphasis to correct rate and depth, listening to Nellie the Elephant as a learning aid during CPR training should be discontinued. Further research is required to identify music that, when played during CPR training, increases the proportion of lay responders providing chest compressions at both the correct rate and depth. PMID- 20008377 TI - Darwin's illness revisited. PMID- 20008379 TI - Personal perspectives in the life sciences for the Royal Society's 350th anniversary. PMID- 20008378 TI - Perceived age as clinically useful biomarker of ageing: cohort study. AB - OBJECTIVE: To determine whether perceived age correlates with survival and important age related phenotypes. DESIGN: Follow-up study, with survival of twins determined up to January 2008, by which time 675 (37%) had died. SETTING: Population based twin cohort in Denmark. PARTICIPANTS: 20 nurses, 10 young men, and 11 older women (assessors); 1826 twins aged >or=70. MAIN OUTCOME MEASURES: Assessors: perceived age of twins from photographs. Twins: physical and cognitive tests and molecular biomarker of ageing (leucocyte telomere length). RESULTS: For all three groups of assessors, perceived age was significantly associated with survival, even after adjustment for chronological age, sex, and rearing environment. Perceived age was still significantly associated with survival after further adjustment for physical and cognitive functioning. The likelihood that the older looking twin of the pair died first increased with increasing discordance in perceived age within the twin pair-that is, the bigger the difference in perceived age within the pair, the more likely that the older looking twin died first. Twin analyses suggested that common genetic factors influence both perceived age and survival. Perceived age, controlled for chronological age and sex, also correlated significantly with physical and cognitive functioning as well as with leucocyte telomere length. CONCLUSION: Perceived age-which is widely used by clinicians as a general indication of a patient's health-is a robust biomarker of ageing that predicts survival among those aged >or=70 and correlates with important functional and molecular ageing phenotypes. PMID- 20008380 TI - Nature's role in sustaining economic development. AB - In this paper, I formalize the idea of sustainable development in terms of intergenerational well-being. I then sketch an argument that has recently been put forward formally to demonstrate that intergenerational well-being increases over time if and only if a comprehensive measure of wealth per capita increases. The measure of wealth includes not only manufactured capital, knowledge and human capital (education and health), but also natural capital (e.g. ecosystems). I show that a country's comprehensive wealth per capita can decline even while gross domestic product (GDP) per capita increases and the UN Human Development Index records an improvement. I then use some rough and ready data from the world's poorest countries and regions to show that during the period 1970-2000 wealth per capita declined in South Asia and sub-Saharan Africa, even though the Human Development Index (HDI) showed an improvement everywhere and GDP per capita increased in all places (except in sub-Saharan Africa, where there was a slight decline). I conclude that, as none of the development indicators currently in use is able to reveal whether development has been, or is expected to be, sustainable, national statistical offices and international organizations should now routinely estimate the (comprehensive) wealth of nations. PMID- 20008381 TI - Crossing scales, crossing disciplines: collective motion and collective action in the Global Commons. AB - Two conflicting tendencies can be seen throughout the biological world: individuality and collective behaviour. Natural selection operates on differences among individuals, rewarding those who perform better. Nonetheless, even within this milieu, cooperation arises, and the repeated emergence of multicellularity is the most striking example. The same tendencies are played out at higher levels, as individuals cooperate in groups, which compete with other such groups. Many of our environmental and other global problems can be traced to such conflicts, and to the unwillingness of individual agents to take account of the greater good. One of the great challenges in achieving sustainability will be in understanding the basis of cooperation, and in taking multicellularity to yet a higher level, finding the pathways to the level of cooperation that is the only hope for the preservation of the planet. PMID- 20008383 TI - The ecosystem-service chain and the biological diversity crisis. AB - The losses that are being incurred of the Earth's biological diversity, at all levels, are now staggering. The trend lines for future loss are steeply upward as new adverse drivers of change come into play. The political processes for matching this crisis are now inadequate and the science needs to address this issue are huge and slow to fulfil, even though strong advances have been made. A more integrated approach to evaluating biodiversity in terms that are meaningful to the larger community is needed that can provide understandable metrics of the consequences to society of the losses that are occurring. Greater attention is also needed in forecasting likely diversity-loss scenarios in the near term and strategies for alleviating detrimental consequences. At the international level, the Convention on Biological Diversity must be revisited to make it more powerful to meet the needs that originally motivated its creation. Similarly, at local and regional levels, an ecosystem-service approach to conservation can bring new understanding to the value, and hence the need for protection, of the existing natural capital. PMID- 20008382 TI - Evolutionary dynamics in structured populations. AB - Evolutionary dynamics shape the living world around us. At the centre of every evolutionary process is a population of reproducing individuals. The structure of that population affects evolutionary dynamics. The individuals can be molecules, cells, viruses, multicellular organisms or humans. Whenever the fitness of individuals depends on the relative abundance of phenotypes in the population, we are in the realm of evolutionary game theory. Evolutionary game theory is a general approach that can describe the competition of species in an ecosystem, the interaction between hosts and parasites, between viruses and cells, and also the spread of ideas and behaviours in the human population. In this perspective, we review the recent advances in evolutionary game dynamics with a particular emphasis on stochastic approaches in finite sized and structured populations. We give simple, fundamental laws that determine how natural selection chooses between competing strategies. We study the well-mixed population, evolutionary graph theory, games in phenotype space and evolutionary set theory. We apply these results to the evolution of cooperation. The mechanism that leads to the evolution of cooperation in these settings could be called 'spatial selection': cooperators prevail against defectors by clustering in physical or other spaces. PMID- 20008384 TI - Ecological science and tomorrow's world. AB - Beginning with an outline of uncertainties about the number of species on Earth today, this paper addresses likely causes and consequences of the manifest acceleration in extinction rates over the past few centuries. The ultimate causes are habitat destruction, alien introductions, overexploitation and climate change. Increases in human numbers and per capita impacts underlie all of these. Against a background review of these factors, I conclude with a discussion of the policy implications for equitably proportionate actions-and of the difficulties in achieving them. PMID- 20008385 TI - Linking biodiversity and ecosystems: towards a unifying ecological theory. AB - Community ecology and ecosystem ecology provide two perspectives on complex ecological systems that have largely complementary strengths and weaknesses. Merging the two perspectives is necessary both to ensure continued scientific progress and to provide society with the scientific means to face growing environmental challenges. Recent research on biodiversity and ecosystem functioning has contributed to this goal in several ways. By addressing a new question of high relevance for both science and society, by challenging existing paradigms, by tightly linking theory and experiments, by building scientific consensus beyond differences in opinion, by integrating fragmented disciplines and research fields, by connecting itself to other disciplines and management issues, it has helped transform ecology not only in content, but also in form. Creating a genuine evolutionary ecosystem ecology that links the evolution of species traits at the individual level, the dynamics of species interactions, and the overall functioning of ecosystems would give new impetus to this much-needed process of unification across ecological disciplines. Recent community evolution models are a promising step in that direction. PMID- 20008386 TI - Food security: contributions from science to a new and greener revolution. AB - There is an intrinsic link between the challenge we face to ensure food security through the twenty-first century and other global issues, most notably climate change, population growth and the need to sustainably manage the world's rapidly growing demand for energy and water. Our progress in reducing global poverty and achieving the Millennium Development Goals will be determined to a great extent by how coherently these long-term challenges are tackled. A key question is whether we can feed a future nine billion people equitably, healthily and sustainably. Science and technology can make a major contribution, by providing practical solutions. Securing this contribution requires that high priority be attached both to research and to facilitating the real world deployment of existing and emergent technologies. Put simply, we need a new, 'greener revolution'. Important areas for focus include: crop improvement; smarter use of water and fertilizers; new pesticides and their effective management to avoid resistance problems; introduction of novel non-chemical approaches to crop protection; reduction of post-harvest losses; and more sustainable livestock and marine production. Techniques and technologies from many disciplines, ranging from biotechnology and engineering to newer fields such as nanotechnology, will be needed. PMID- 20008387 TI - Understanding and using quantitative genetic variation. AB - Quantitative genetics, or the genetics of complex traits, is the study of those characters which are not affected by the action of just a few major genes. Its basis is in statistical models and methodology, albeit based on many strong assumptions. While these are formally unrealistic, methods work. Analyses using dense molecular markers are greatly increasing information about the architecture of these traits, but while some genes of large effect are found, even many dozens of genes do not explain all the variation. Hence, new methods of prediction of merit in breeding programmes are again based on essentially numerical methods, but incorporating genomic information. Long-term selection responses are revealed in laboratory selection experiments, and prospects for continued genetic improvement are high. There is extensive genetic variation in natural populations, but better estimates of covariances among multiple traits and their relation to fitness are needed. Methods based on summary statistics and predictions rather than at the individual gene level seem likely to prevail for some time yet. PMID- 20008388 TI - Fluctuating selection: the perpetual renewal of adaptation in variable environments. AB - Darwin insisted that evolutionary change occurs very slowly over long periods of time, and this gradualist view was accepted by his supporters and incorporated into the infinitesimal model of quantitative genetics developed by R. A. Fisher and others. It dominated the first century of evolutionary biology, but has been challenged in more recent years both by field surveys demonstrating strong selection in natural populations and by quantitative trait loci and genomic studies, indicating that adaptation is often attributable to mutations in a few genes. The prevalence of strong selection seems inconsistent, however, with the high heritability often observed in natural populations, and with the claim that the amount of morphological change in contemporary and fossil lineages is independent of elapsed time. I argue that these discrepancies are resolved by realistic accounts of environmental and evolutionary changes. First, the physical and biotic environment varies on all time-scales, leading to an indefinite increase in environmental variance over time. Secondly, the intensity and direction of natural selection are also likely to fluctuate over time, leading to an indefinite increase in phenotypic variance in any given evolving lineage. Finally, detailed long-term studies of selection in natural populations demonstrate that selection often changes in direction. I conclude that the traditional gradualist scheme of weak selection acting on polygenic variation should be supplemented by the view that adaptation is often based on oligogenic variation exposed to commonplace, strong, fluctuating natural selection. PMID- 20008389 TI - Understanding plant reproductive diversity. AB - Flowering plants display spectacular floral diversity and a bewildering array of reproductive adaptations that promote mating, particularly outbreeding. A striking feature of this diversity is that related species often differ in pollination and mating systems, and intraspecific variation in sexual traits is not unusual, especially among herbaceous plants. This variation provides opportunities for evolutionary biologists to link micro-evolutionary processes to the macro-evolutionary patterns that are evident within lineages. Here, I provide some personal reflections on recent progress in our understanding of the ecology and evolution of plant reproductive diversity. I begin with a brief historical sketch of the major developments in this field and then focus on three of the most significant evolutionary transitions in the reproductive biology of flowering plants: the pathway from outcrossing to predominant self-fertilization, the origin of separate sexes (females and males) from hermaphroditism and the shift from animal pollination to wind pollination. For each evolutionary transition, I consider what we have discovered and some of the problems that still remain unsolved. I conclude by discussing how new approaches might influence future research in plant reproductive biology. PMID- 20008390 TI - Deep phylogeny, ancestral groups and the four ages of life. AB - Organismal phylogeny depends on cell division, stasis, mutational divergence, cell mergers (by sex or symbiogenesis), lateral gene transfer and death. The tree of life is a useful metaphor for organismal genealogical history provided we recognize that branches sometimes fuse. Hennigian cladistics emphasizes only lineage splitting, ignoring most other major phylogenetic processes. Though methodologically useful it has been conceptually confusing and harmed taxonomy, especially in mistakenly opposing ancestral (paraphyletic) taxa. The history of life involved about 10 really major innovations in cell structure. In membrane topology, there were five successive kinds of cell: (i) negibacteria, with two bounding membranes, (ii) unibacteria, with one bounding and no internal membranes, (iii) eukaryotes with endomembranes and mitochondria, (iv) plants with chloroplasts and (v) finally, chromists with plastids inside the rough endoplasmic reticulum. Membrane chemistry divides negibacteria into the more advanced Glycobacteria (e.g. Cyanobacteria and Proteobacteria) with outer membrane lipolysaccharide and primitive Eobacteria without lipopolysaccharide (deserving intenser study). It also divides unibacteria into posibacteria, ancestors of eukaryotes, and archaebacteria-the sisters (not ancestors) of eukaryotes and the youngest bacterial phylum. Anaerobic eobacteria, oxygenic cyanobacteria, desiccation-resistant posibacteria and finally neomura (eukaryotes plus archaebacteria) successively transformed Earth. Accidents and organizational constraints are as important as adaptiveness in body plan evolution. PMID- 20008391 TI - Evolution: like any other science it is predictable. AB - Evolutionary biology rejoices in the diversity of life, but this comes at a cost: other than working in the common framework of neo-Darwinian evolution, specialists in, for example, diatoms and mammals have little to say to each other. Accordingly, their research tends to track the particularities and peculiarities of a given group and seldom enquires whether there are any wider or deeper sets of explanations. Here, I present evidence in support of the heterodox idea that evolution might look to a general theory that does more than serve as a tautology ('evolution explains evolution'). Specifically, I argue that far from its myriad of products being fortuitous and accidental, evolution is remarkably predictable. Thus, I urge a move away from the continuing obsession with Darwinian mechanisms, which are entirely uncontroversial. Rather, I emphasize why we should seek explanations for ubiquitous evolutionary convergence, as well as the emergence of complex integrated systems. At present, evolutionary theory seems to be akin to nineteenth-century physics, blissfully unaware of the imminent arrival of quantum mechanics and general relativity. Physics had its Newton, biology its Darwin: evolutionary biology now awaits its Einstein. PMID- 20008392 TI - The new biology of ageing. AB - Human life expectancy in developed countries has increased steadily for over 150 years, through improvements in public health and lifestyle. More people are hence living long enough to suffer age-related loss of function and disease, and there is a need to improve the health of older people. Ageing is a complex process of damage accumulation, and has been viewed as experimentally and medically intractable. This view has been reinforced by the realization that ageing is a disadvantageous trait that evolves as a side effect of mutation accumulation or a benefit to the young, because of the decline in the force of natural selection at later ages. However, important recent discoveries are that mutations in single genes can extend lifespan of laboratory model organisms and that the mechanisms involved are conserved across large evolutionary distances, including to mammals. These mutations keep the animals functional and pathology-free to later ages, and they can protect against specific ageing-related diseases, including neurodegenerative disease and cancer. Preliminary indications suggest that these new findings from the laboratory may well also apply to humans. Translating these discoveries into medical treatments poses new challenges, including changing clinical thinking towards broad-spectrum, preventative medicine and finding novel routes to drug development. PMID- 20008393 TI - The therapeutic potential of stem cells. AB - In recent years, there has been an explosion of interest in stem cells, not just within the scientific and medical communities but also among politicians, religious groups and ethicists. Here, we summarize the different types of stem cells that have been described: their origins in embryonic and adult tissues and their differentiation potential in vivo and in culture. We review some current clinical applications of stem cells, highlighting the problems encountered when going from proof-of-principle in the laboratory to widespread clinical practice. While some of the key genetic and epigenetic factors that determine stem cell properties have been identified, there is still much to be learned about how these factors interact. There is a growing realization of the importance of environmental factors in regulating stem cell behaviour and this is being explored by imaging stem cells in vivo and recreating artificial niches in vitro. New therapies, based on stem cell transplantation or endogenous stem cells, are emerging areas, as is drug discovery based on patient-specific pluripotent cells and cancer stem cells. What makes stem cell research so exciting is its tremendous potential to benefit human health and the opportunities for interdisciplinary research that it presents. PMID- 20008394 TI - The social brain: allowing humans to boldly go where no other species has been. AB - The biological basis of complex human social interaction and communication has been illuminated through a coming together of various methods and disciplines. Among these are comparative studies of other species, studies of disorders of social cognition and developmental psychology. The use of neuroimaging and computational models has given weight to speculations about the evolution of social behaviour and culture in human societies. We highlight some networks of the social brain relevant to two-person interactions and consider the social signals between interacting partners that activate these networks. We make a case for distinguishing between signals that automatically trigger interaction and cooperation and ostensive signals that are used deliberately. We suggest that this ostensive signalling is needed for 'closing the loop' in two-person interactions, where the partners each know that they have the intention to communicate. The use of deliberate social signals can serve to increase reputation and trust and facilitates teaching. This is likely to be a critical factor in the steep cultural ascent of mankind. PMID- 20008395 TI - Learning to represent visual input. AB - One of the central problems in computational neuroscience is to understand how the object-recognition pathway of the cortex learns a deep hierarchy of nonlinear feature detectors. Recent progress in machine learning shows that it is possible to learn deep hierarchies without requiring any labelled data. The feature detectors are learned one layer at a time and the goal of the learning procedure is to form a good generative model of images, not to predict the class of each image. The learning procedure only requires the pairwise correlations between the activations of neuron-like processing units in adjacent layers. The original version of the learning procedure is derived from a quadratic 'energy' function but it can be extended to allow third-order, multiplicative interactions in which neurons gate the pairwise interactions between other neurons. A technique for factoring the third-order interactions leads to a learning module that again has a simple learning rule based on pairwise correlations. This module looks remarkably like modules that have been proposed by both biologists trying to explain the responses of neurons and engineers trying to create systems that can recognize objects. PMID- 20008397 TI - Sequences and consequences. AB - The conversion of data into knowledge constitutes a great challenge for future biological research. The new science of Systems Biology claims to be able to solve the problem but I contend that this approach will fail because deducing models of function from the behaviour of a complex system is an inverse problem that is impossible to solve. In addition, one cannot easily escape into high level holistic approaches, since the essence of all biological systems is that they are encoded as molecular descriptions in their genes and since genes are molecules and exert their functions through other molecules, the molecular explanation must constitute the core of understanding biological systems. We then solve the forward problem of computing the behaviour of the system from its components and their interactions. I propose that the correct level of abstraction is the cell and provide an outline of CELLMAP, a design for a system to organize biological information. PMID- 20008396 TI - Genome-wide scans for footprints of natural selection. AB - Detecting recent selected 'genomic footprints' applies directly to the discovery of disease genes and in the imputation of the formative events that molded modern population genetic structure. The imprints of historic selection/adaptation episodes left in human and animal genomes allow one to interpret modern and ancestral gene origins and modifications. Current approaches to reveal selected regions applied in genome-wide selection scans (GWSSs) fall into eight principal categories: (I) phylogenetic footprinting, (II) detecting increased rates of functional mutations, (III) evaluating divergence versus polymorphism, (IV) detecting extended segments of linkage disequilibrium, (V) evaluating local reduction in genetic variation, (VI) detecting changes in the shape of the frequency distribution (spectrum) of genetic variation, (VII) assessing differentiating between populations (F(ST)), and (VIII) detecting excess or decrease in admixture contribution from one population. Here, we review and compare these approaches using available human genome-wide datasets to provide independent verification (or not) of regions found by different methods and using different populations. The lessons learned from GWSSs will be applied to identify genome signatures of historic selective pressures on genes and gene regions in other species with emerging genome sequences. This would offer considerable potential for genome annotation in functional, developmental and evolutionary contexts. PMID- 20008398 TI - An examination of technical efficiency, quality, and patient safety in acute care nursing units. AB - Using an innovative statistical approach-data envelopment analysis-the authors examined the technical efficiency of 226 medical, surgical, and medical-surgical nursing units in 118 randomly selected acute care hospitals. The authors used the inputs of registered nurse, licensed practical nurse, and unlicensed hours of care; operating expenses; and number of beds on the unit. Outputs included case mix adjusted discharges, patient satisfaction (as a quality measure), and the rates of medication errors and patient falls (as measures of patient safety). This study found that 60% of units were operating at less than full efficiency. Key areas for improvement included slight reductions in labor hours and large reductions in medication errors and falls. The study findings indicate the importance of improving patient safety as a mechanism to simultaneously improve nursing unit efficiency. PMID- 20008399 TI - Signal mechanisms of vascular endothelial growth factor and interleukin-8 in ovarian hyperstimulation syndrome: dopamine targets their common pathways. AB - BACKGROUND: Ovarian hyperstimulation syndrome (OHSS) is a serious complication of ovarian stimulation with massive ascites, pleural effusion and hemoconcentration. The pathophysiological signal mechanisms of OHSS are still unclear and merit further investigation. METHODS: Various angiogenic cytokines of follicular fluid and ascites of patients with risk of OHSS were measured, and examined for inducing endothelial permeability. These include vascular endothelial growth factor (VEGF), interleukin (IL)-6, IL-8, basic fibroblast growth factor, tumor necrosis factor-alpha, IL-1alpha, IL-1beta and platelet-derived growth factor. We explore the molecular signal pathways of major contributing cytokines in granulosa-lutein cells and endothelial cells possibly involved in OHSS. RESULTS: Neutralizing antibodies of VEGF or IL-8 significantly decreased follicular fluid- and ascites-induced endothelial permeability. Human chorionic gonadotrophin induced VEGF secretion of granulosa-lutein cells through the Sp1 and CREB dependent pathways. IL-8 activated CXCR1/2 of endothelial cells leading to VEGF receptor (VEGFR)-2 transactivation. Both VEGF and IL-8 of follicular fluid enhanced endothelial permeability via VEGFR-2-mediated Rho/Rock activation, actin polymerization and phosphorylations of VE-cadherin and occludin, resulting in opening of adherens junctions and tight junctions. Dopamine (2 microM) inhibited follicular fluid-induced VEGFR-2 signals and endothelial permeability, without diminishing migration and tube formation. CONCLUSIONS: Our results suggest that VEGF and IL-8 secreted from corpora luteae may play major roles in OHSS. Delineation of signal pathways would be helpful for treatment. Dopamine may block VEGF- and IL-8-induced endothelial permeability by inhibiting common VEGFR-2 dependent signals. PMID- 20008401 TI - Is cocoa good for the heart? PMID- 20008402 TI - Delayed complex systems: an overview. PMID- 20008400 TI - Gene-centered regulatory networks. AB - Differential gene expression plays a critical role in the development and physiology of multicellular organisms. At a 'systems level' (e.g. at the level of a tissue, organ or whole organism), this process can be studied using gene regulatory network (GRN) models that capture physical and regulatory interactions between genes and their regulators. In the past years, significant progress has been made toward the mapping of GRNs using a variety of experimental and computational approaches. Here, we will discuss gene-centered approaches that we employed to characterize GRNs and describe insights that we have obtained into the global design principles of gene regulation in complex metazoan systems. PMID- 20008403 TI - Extending anticipation horizon of chaos synchronization schemes with time-delay coupling. AB - We analyse anticipating synchronization in chaotic systems with time-delay coupling. Two algorithms for extending the prediction horizon are considered. One of them is based on the design of a suitable coupling matrix compensating the phase lag in the time-delay feedback term of the slave system. The second algorithm extends the first by incorporating, in the coupling law, information from many previous states of the master and slave systems. We demonstrate the efficiency of both algorithms with the simple dynamical model of coupled unstable spirals, as well as with the coupled Rossler systems. The maximum prediction time attained for the Rossler system is equal to the characteristic period of chaotic oscillations. PMID- 20008404 TI - Delay stabilization of periodic orbits in coupled oscillator systems. AB - We study diffusively coupled oscillators in Hopf normal form. By introducing a non-invasive delay coupling, we are able to stabilize the inherently unstable anti-phase orbits. For the super- and subcritical cases, we state a condition on the oscillator's nonlinearity that is necessary and sufficient to find coupling parameters for successful stabilization. We prove these conditions and review previous results on the stabilization of odd-number orbits by time-delayed feedback. Finally, we illustrate the results with numerical simulations. PMID- 20008405 TI - Complex dynamics and synchronization of delayed-feedback nonlinear oscillators. AB - We describe a flexible and modular delayed-feedback nonlinear oscillator that is capable of generating a wide range of dynamical behaviours, from periodic oscillations to high-dimensional chaos. The oscillator uses electro-optic modulation and fibre-optic transmission, with feedback and filtering implemented through real-time digital signal processing. We consider two such oscillators that are coupled to one another, and we identify the conditions under which they will synchronize. By examining the rates of divergence or convergence between two coupled oscillators, we quantify the maximum Lyapunov exponents or transverse Lyapunov exponents of the system, and we present an experimental method to determine these rates that does not require a mathematical model of the system. Finally, we demonstrate a new adaptive control method that keeps two oscillators synchronized, even when the coupling between them is changing unpredictably. PMID- 20008406 TI - Quantifying stochasticity in the dynamics of delay-coupled semiconductor lasers via forbidden patterns. AB - We quantify the level of stochasticity in the dynamics of two mutually coupled semiconductor lasers. Specifically, we concentrate on a regime in which the lasers synchronize their dynamics with a non-zero lag time, and the leader and laggard roles alternate irregularly between the lasers. We analyse this switching dynamics in terms of the number of forbidden patterns of the alternate time series. The results reveal that the system operates in a stochastic regime, with the level of stochasticity decreasing as the lasers are pumped further away from their lasing threshold. This behaviour is similar to that exhibited by a single semiconductor laser subject to external optical feedback, as its dynamics shifts from the regime of low-frequency fluctuations to coherence collapse. PMID- 20008407 TI - On chaos synchronization and secure communication. AB - Chaos synchronization, in particular isochronal synchronization of two chaotic trajectories to each other, may be used to build a means of secure communication over a public channel. In this paper, we give an overview of coupling schemes of Bernoulli units deduced from chaotic laser systems, different ways to transmit information by chaos synchronization and the advantage of bidirectional over unidirectional coupling with respect to secure communication. We present the protocol for using dynamical private commutative filters for tap-proof transmission of information that maps the task of a passive attacker to the class of non-deterministic polynomial time-complete problems. PMID- 20008408 TI - Interplay of time-delayed feedback control and temporally correlated noise in excitable systems. AB - The interplay of time-delayed feedback and temporally correlated coloured noise in a single and two coupled excitable systems is studied in the framework of the FitzHugh-Nagumo (FHN) model. By using coloured noise instead of white noise, the noise correlation time is introduced as an additional time scale. We show that in a single FHN system the major time scale of oscillations is strongly influenced by the noise correlation time, which in turn affects the maxima of coherence with respect to the delay time. In two coupled FHN systems, coloured noise input to one subsystem influences coherence resonance and stochastic synchronization of both subsystems. Application of delayed feedback control to the coloured noise driven subsystem is shown to change coherence and time scales of noise-induced oscillations in both systems, and to enhance or suppress stochastic synchronization under certain conditions. PMID- 20008409 TI - Stochastic two-delay differential model of delayed visual feedback effects on postural dynamics. AB - We report on experiments and modelling involving the 'visuo-postural control loop' in the upright stance. We experimentally manipulated an artificial delay to the visual feedback during standing, presented at delays ranging from 0 to 1 s in increments of 250 ms. Using stochastic delay differential equations, we explicitly modelled the centre-of-pressure (COP) and centre-of-mass (COM) dynamics with two independent delay terms for vision and proprioception. A novel 'drifting fixed point' hypothesis was used to describe the fluctuations of the COM with the COP being modelled as a faster, corrective process of the COM. The model was in good agreement with the data in terms of probability density functions, power spectral densities, short- and long-term correlations (Hurst exponents) as well the critical time between the two ranges. PMID- 20008410 TI - Controlling biological networks by time-delayed signals. AB - This paper describes the use of time-delayed feedback to regulate the behaviour of biological networks. The general ideas on specific transcriptional regulatory and neural networks are demonstrated. It is shown that robust yet tunable controllers can be constructed that provide the biological systems with model engineered inputs. The results indicate that time delay modulation may serve as an efficient biocompatible control tool. PMID- 20008411 TI - Oscillatory response in a sensory network of ON and OFF cells with instantaneous and delayed recurrent connections. AB - A neural field model with multiple cell-to-cell feedback connections is investigated. Our model incorporates populations of ON and OFF cells, receiving sensory inputs with direct and inverted polarity, respectively. Oscillatory responses to spatially localized stimuli are found to occur via Andronov-Hopf bifurcations of stationary activity. We explore the impact of multiple delayed feedback components as well as additional excitatory and/or inhibitory non delayed recurrent signals on the instability threshold. Paradoxically, instantaneous excitatory recurrent terms are found to enhance network responsiveness by reducing the oscillatory response threshold, allowing smaller inputs to trigger oscillatory activity. Instantaneous inhibitory components do the opposite. The frequency of these response oscillations is further shaped by the polarity of the non-delayed terms. PMID- 20008412 TI - Delayed feedback of sampled higher derivatives. AB - Systems where the present rate of change of the state depends on the past values of the higher rates of change of the state are described by so-called advanced functional differential equations (AFDEs). In an AFDE, the highest derivative of the state-space coordinate appears with delayed argument only. The corresponding linearized equations are always unstable with infinitely many unstable poles, and are rarely related to practical applications due to their inherently implicit nature. In this paper, one of the simplest AFDEs, a linear scalar first-order system, is considered with the delayed feedback of the second derivative of the state in the presence of sampling in the feedback loop (i.e. in the case of digital control). It is shown that sampling of the feedback may stabilize the originally infinitely unstable system for certain parameter combinations. The result explains the stable behaviour of certain dynamical systems with feedback delay in the highest derivative. PMID- 20008413 TI - Bifurcation to fronts due to delay. AB - The stability of a steady-state front (kink) subject to a time-delayed feedback control (TDFC) is examined in detail. TDFC is based on the use of the difference between system variables at the current moment of time and their values at some time in the past. We first show that there exists a bifurcation to a moving front. We then investigate the limit of large delays but weak feedback and obtain a global bifurcation diagram for the propagation speed. Finally, we examine the case of a two-dimensional front with radial symmetry and determine the critical radius above which propagation is possible. PMID- 20008414 TI - On the origin of chaos in autonomous Boolean networks. AB - We undertake a systematic study of the dynamics of Boolean networks to determine the origin of chaos observed in recent experiments. Networks with nodes consisting of ideal logic gates are known to display either steady states, periodic behaviour or an ultraviolet catastrophe where the number of logic transition events circulating in the network per unit time grows as a power law. In an experiment, the non-ideal behaviour of the logic gates prevents the ultraviolet catastrophe and may lead to deterministic chaos. We identify certain non-ideal features of real logic gates that enable chaos in experimental networks. We find that short-pulse rejection and asymmetry between the logic states tend to engender periodic behaviour, at least for the simplest networks. On the other hand, we find that a memory effect termed 'degradation' can generate chaos. Our results strongly suggest that deterministic chaos can be expected in a large class of experimental Boolean-like networks. Such devices may find application in a variety of technologies requiring fast complex waveforms or flat power spectra, and can be used as a test-bed for fundamental studies of real world Boolean-like networks. PMID- 20008415 TI - Proteomic analysis of endometrium from fertile and infertile patients suggests a role for apolipoprotein A-I in embryo implantation failure and endometriosis. AB - Pregnancy is dependent upon the endometrium acquiring a receptive phenotype that facilitates apposition, adhesion and invasion of a developmentally competent embryo. Surface-enhanced laser desorption/ionization time-of-flight mass spectrometry of mid-secretory endometrial biopsies revealed a 28 kDa protein peak that discriminated highly between samples obtained from women with recurrent implantation failure and fertile controls. Subsequent tandem mass spectroscopy unambiguously identified this peak as apolipoprotein A-I (apoA-I), a potent anti inflammatory molecule. Total endometrial apoA-I levels were, however, comparable between the study and control group. Moreover, endometrial apoA-I mRNA expression was not cycle-dependent although there was partial loss of apoA-I immunoreactivity in luminal and glandular epithelium in mid-secretory compared with proliferative endometrial samples. Because of its putative anti-implantation properties, we examined whether endometrial apoA-I expression is regulated by embryonic signals. Human chorionic gonadotrophin (hCG) strongly inhibited apoA-I expression in differentiating explant cultures but not when established from eutopic endometrium from patients with endometriosis. Pelvic endometriosis was associated with elevated apoA-I mRNA levels, increased secretion by differentiating eutopic endometrial explant cultures and lack of hCG-dependent down-regulation. To corroborate these observations, we examined endometrial apoA I expression and its regulation by hCG in a non-human primate model of endometriosis. As in humans, hCG strongly inhibited endometrial apoA-I mRNA expression in disease-free baboons, but this response was entirely lost upon induction of pelvic endometriosis. Together, these observations indicate that perturbations in endometrial apoA-I expression, modification or regulation by paracrine embryonic signals play a major role in implantation failure and infertility. PMID- 20008416 TI - Clinical utility of Rome criteria managing functional gastrointestinal disorders in pediatric primary care. AB - OBJECTIVE: Our goals were to demonstrate that functional gastrointestinal disorders (FGIDs) can be diagnosed in a positive fashion and managed by family pediatricians (FPs); to assess the compliance of FPs with a predefined diagnostic/therapeutic protocol for managing FGIDs to evaluate efficacy of continuing medical education; and to evaluate the success of reassurance by using a biopsychosocial model in comparison to drug treatment in an open-label, nonrandomized study. PATIENTS AND METHODS: A total of 9291 patients, aged birth to 14 years, were prospectively enrolled during a 3-month period by 21 FPs. Follow-up was performed at 1-, 3-, and 12-month intervals. To assess compliance, after diagnosis, FPs that participated in the study were asked to comply with the diagnostic/therapeutic protocol and record investigations and treatment prescribed. RESULTS: Two hundred sixty-one patients (2.8%) met Rome II criteria and were included in the study. In all cases but 4, diagnosis of FGIDs was confirmed at the end of follow-up (98.4%). Average compliance of FPs was >80%. Among 56 patients treated only with the explanation of symptom and reassurance, 52 (92.8%) have reported success, in comparison with 26 of 35 patients (74.3%) treated with drugs (odds ratio: 4.5 [95% confidence interval: 1.3-16]). CONCLUSIONS: FGIDs can be diagnosed and managed in primary care. Prospective studies in primary care allow assessment of compliance of pediatricians and implementation of what is learned in courses developed in continuing medical education. Informed reassurance and availability of FPs are more useful than over the-counter drugs, which are often used for FGIDs. PMID- 20008417 TI - Hearing impairment in childhood bacterial meningitis is little relieved by dexamethasone or glycerol. AB - OBJECTIVE: Several studies have evaluated dexamethasone for prevention of hearing loss in childhood bacterial meningitis, but results have varied. We compared dexamethasone and/or glycerol recipients with placebo recipients, and measured hearing at 3 threshold levels. METHODS: Children aged 2 months to 16 years with meningitis were treated with ceftriaxone but were double-blindly randomly assigned to receive adjuvant dexamethasone intravenously, glycerol orally, both agents, or neither agent. We used the Glasgow coma scale to grade the presenting status. The end points were the better ear's ability to detect sounds of >40 dB, >or=60 dB, and >or=80 dB, with these thresholds indicating any, moderate-to severe, or severe impairment, respectively. All tests were interpreted by an external audiologist. Influence of covariates in the treatment groups was examined by binary logistic regression. RESULTS: Of the 383 children, mostly with meningitis caused by Haemophilus influenzae type b or Streptococcus pneumoniae, 101 received dexamethasone, 95 received dexamethasone and glycerol, 92 received glycerol, and 95 received placebo. Only the presenting condition and young age predicted impairment independently through all threshold levels. Each lowering point in the Glasgow scale increased the risk by 15% to 21% (odds ratio: 1.20, 1.21, and 1.15 [95% confidence interval: 1.06-1.35, 1.07-1.37, and 1.01-1.31]; P = .005, .003, and .039) for any, moderate-to-severe, or severe impairment, respectively. Each increasing month of age decreased the risk by 2% to 6% (P = .0001, .0007, and .041, respectively). Neither dexamethasone nor glycerol prevented hearing loss at these levels regardless of the causative agent or timing of antimicrobial agent. CONCLUSIONS: With bacterial meningitis, the child's presenting status and young age are the most important predictors of hearing impairment. Little relief is obtained from current adjuvant medications. PMID- 20008418 TI - Emergency department reliance: a discriminatory measure of frequent emergency department users. AB - OBJECTIVE: High emergency department (ED) use has previously been defined as a person's having a large number of ED visits, implying that all frequent users are the same. ED reliance (EDR), the percentage of all health care visits that occur in the ED, considers ED use in relation to primary care use and, thus, may discriminate among high-ED-user populations. Our objective was to determine whether EDR, as a complementary use measure, could differentiate frequent users secondary to increased need for care from those with access issues. METHODS: We conducted an analysis of prospectively collected data from the Medical Expenditure Panel Survey from 2000-2001 and 2001-2002. Frequent ED users were defined as having >or=2 ED visits, and EDR was dichotomized as high (>0.33) or low (or=50 years. AB - OBJECTIVE: This study examined patterns of work and retirement among older pediatricians, including the determinants of part-time work and retirement, as well as extended participation in the pediatrician workforce. METHODS: A mail survey regarding recent and expected workforce participation was distributed to American Academy of Pediatrics members who were aged >or=50 years. A total of 1158 (72%) responded, 1114 (70%) of whom were included in these analyses. Analyses examined levels of engagement in medicine, retirement plans, and differences between men and women and primary care pediatricians and pediatric subspecialists. RESULTS: Providing or planning to provide patient care past age 65 was reported by 45% of the respondents. Engaging in part-time work was increasingly more common with age and may represent a step toward retirement for many. In this sample, women were more likely to work part-time (32% vs 18% of men) and less likely to work past age 65 (26% vs 57% of men). When compared with primary care pediatricians, pediatric subspecialists worked more hours per week (59 vs 53) but spent a smaller percentage of their time on patient care (63% vs 82%). CONCLUSIONS: Part-time work and reduced work hours in anticipation of retirement are options that are used and desired by older pediatricians. Results of this study suggest that making provisions for gradual reduction in work hours or other forms of phasing out of the workforce could benefit the practice of pediatrics by extending the career length of the most experienced pediatricians. PMID- 20008421 TI - 2009 Joseph W. St Geme Jr Leadership Award Address: the whole--and the sum of its parts. PMID- 20008422 TI - Resective pediatric epilepsy surgery in Lennox-Gastaut syndrome. AB - OBJECTIVE: The objective of this study was to evaluate the role of resective pediatric epilepsy surgery for Lennox-Gastaut syndrome (LGS). METHODS: We analyzed clinical data of 27 children and adolescents who had LGS and underwent resective epilepsy surgery despite abundant (>30% of preoperative interictal and/or ictal epileptiform discharges) generalized or generalized contralateral maximal and multiregional electroencephalogram abnormalities. RESULTS: On high resolution MRI, cerebral lesions were noted in 23 (85.2%) patients but not in 4 (14.8%) patients. The age of patients at the time of surgery was between 1.7 and 17.3 years (mean: 7.8 years). Surgeries were lobar or multilobar resection in 21 (77.8%) patients and hemispherotomy in 6 (22.2%). At a mean of 33.1 months' postoperative follow-up, 16 (59.3%) patients had no seizures and 4 (14.8%) had infrequent seizures. Of 4 patients without brain abnormalities found on MRI, 2 patients became seizure-free after resective surgery was performed on the basis of electrophysiologic studies and concordant results in other multimodal neuroimages. Malformation of cortical development was the most common pathology and was seen in 20 (74.1%) patients, but 2 (7.4% patients) did not show any abnormal pathology. Sixteen (72.7%) patients, including 14 who had no seizures and 2 who had infrequent seizures after surgery, showed an increase in developmental quotient. No clinical profile was significantly associated with postoperative seizure-free rate. CONCLUSIONS: Resective epilepsy surgery should be considered for children with LGS, despite abundant generalized and multiregional electroencephalogram abnormalities. PMID- 20008424 TI - The choking game: physician perspectives. AB - OBJECTIVE: The goal was to assess awareness of the choking game among physicians who care for adolescents and to explore their opinions regarding its inclusion in anticipatory guidance. METHODS: We surveyed 865 pediatricians and family practitioners. The survey was designed to assess physicians' awareness of the choking game and its warning signs, the suspected prevalence of patients' participation in the activity, and the willingness of physicians to include the choking game in adolescent anticipatory guidance. Information on the general use of anticipatory guidance also was collected. RESULTS: The survey was completed by 163 physicians (response rate: 21.8%). One-hundred eleven (68.1%) had heard of the choking game, 68 of them (61.3%) through sources in the popular media. General pediatricians were significantly more likely to report being aware of the choking game than were family practitioners or pediatric subspecialists (P = .004). Of physicians who were aware of the choking game, 75.7% identified >or=1 warning sign and 52.3% identified >or=3. Only 7.6% of physicians who were aware of the choking game reported that they cared for a patient they suspected was participating in the activity, and 2 (1.9%) reported that they include the choking game in anticipatory guidance for adolescents. However, 64.9% of all respondents agreed that the choking game should be included in anticipatory guidance. CONCLUSIONS: Close to one third of physicians surveyed were unaware of the choking game, a potentially life-threatening activity practiced by adolescents. Despite acknowledging that the choking game should be included in adolescent anticipatory guidance, few physicians reported actually discussing it. To provide better care for their adolescent patients, pediatricians and family practitioners should be knowledgeable about risky behaviors encountered by their patients, including the choking game, and provide timely guidance about its dangers. PMID- 20008425 TI - Effect of domperidone on the composition of preterm human breast milk. AB - OBJECTIVE: Domperidone is increasingly prescribed to improve breast milk volume despite a lack of evidence regarding its effects on breast milk composition. We examined the effect of domperidone on the nutrient composition of breast milk. PATIENTS AND METHODS: Forty-six mothers who had delivered infants at <31 weeks' gestation, who experienced lactation failure, were randomly assigned to receive domperidone or placebo for 14 days. Protein, energy, fat, carbohydrate, sodium, calcium, and phosphate levels in breast milk were measured on days 0, 4, 7, and 14, serum prolactin levels were measured on days 0, 4, and 14, and total milk volume was recorded daily. Mean within-subject changes in nutrients and milk volumes were examined. RESULTS: Maternal and infant characteristics, serum prolactin level, and breast milk volume and composition were not significantly different between domperidone and placebo groups on day 0. By day 14, breast milk volumes increased by 267% in the domperidone-treated group and by 18.5% in the placebo group (P = .005). Serum prolactin increased by 97% in the domperidone group and by 17% in the placebo group (P = .07). Mean breast milk protein declined by 9.6% in the domperidone group and increased by 3.6% in the placebo group (P = .16). Changes in energy, fat, carbohydrate, sodium, and phosphate content were also not significantly different between groups. Significant increases were observed in breast milk carbohydrate (2.7% vs -2.7%; P = .05) and calcium (61.8% vs -4.4%; P = .001) in the domperidone versus placebo groups. No significant adverse events were observed among mothers or infants. CONCLUSION: Domperidone increases the volume of breast milk of preterm mothers experiencing lactation failure, without substantially altering the nutrient composition. PMID- 20008426 TI - Prevalence and treatment of mental disorders among US children in the 2001-2004 NHANES. AB - OBJECTIVE: This article presents the 12-month prevalence estimates of specific mental disorders, their social and demographic correlates, and service use patterns in children and adolescents from the National Health and Nutrition Examination Survey, a nationally representative probability sample of noninstitutionalized US civilians. METHODS: The sample includes 3042 participants 8 to 15 years of age from cross-sectional surveys conducted from 2001 to 2004. Data on Diagnostic and Statistical Manual of Mental Disorders, Fourth Edition criteria for mental disorders were derived from administration of selected modules of the National Institute of Mental Health Diagnostic Interview Schedule for Children, version IV, a structured diagnostic interview administered by lay interviewers to assess psychiatric diagnoses of children and adolescents. RESULTS: Twelve-month prevalence rates of Diagnostic and Statistical Manual of Mental Disorders, Fourth Edition-defined disorders in this sample were 8.6% for attention-deficit/hyperactivity disorder, 3.7% for mood disorders, 2.1% for conduct disorder, 0.7% for panic disorder or generalized anxiety disorder, and 0.1% for eating disorders. Boys had 2.1 times greater prevalence of attention deficit/hyperactivity disorder than girls, girls had twofold higher rates of mood disorders than boys, and there were no gender differences in the rates of anxiety disorders or conduct disorder. Only approximately one half of those with one of the disorders assessed had sought treatment with a mental health professional. CONCLUSION: These data constitute a first step in building a national database on mental health in children and adolescents. PMID- 20008427 TI - Slower reaction times and impaired learning in young adults with birth weight <1500 g. AB - OBJECTIVE: Children with very low birth weight (VLBW; <1500 g) perform worse on cognitive tests than do children who are born at term. Whether this difference persists into adulthood has been little studied. We assessed core neurocognitive abilities (processing speed, working memory, attention, and learning capacity) in young adults with VLBW and in term-born control subjects. METHODS: In conjunction with the Helsinki Study of Very Low Birth Weight Adults, 147 VLBW and 171 control subjects who were aged 18 to 27 years and did not have neurosensory impairments performed a computerized test battery (CogState Ltd, Melbourne, Australia). T tests and linear regression models were used. Cohen's d was used to express effect size (ES). RESULTS: VLBW adults had slower reaction times than did control subjects on all 5 tasks: simple reaction time (mean difference: 4.0% [95% confidence interval (CI): 1.1%-7.0%]; ES: 0.30), choice reaction time (mean difference: 3.2% [95% CI: 0.3%-6.2%]; ES: 0.24), working memory (mean difference: 8.4% [95% CI: 3.7%-13.4%]; ES: 0.40), divided attention (mean difference: 7.2% [95% CI: 2.7%-11.9%]; ES: 0.36), and associated learning reaction time (mean difference: 6.4% [95% CI: 1.3%-11.9%]; ES: 0.28). In addition, VLBW adults showed impaired learning abilities on the associated learning task (percentage of correct responses: 85.7 vs 80.2; P < .001; ES: 0.64). The results were little affected by adjustment for confounders. CONCLUSIONS: Nonimpaired VLBW individuals exhibited slower psychomotor speed and lower accuracy on the associated learning task. These results indicate that very preterm birth, even when obvious neurosensory deficits are absent, may have long-term consequences on core neurocognitive abilities. PMID- 20008428 TI - Impact of the fast track prevention program on health services use by conduct problem youth. AB - OBJECTIVE: We tested the impact of the Fast Track conduct disorder prevention program on the use of pediatric, general health, and mental health services in adolescence. PATIENTS AND METHODS: Participants were 891 public kindergarten boys and girls screened from a population of 9594 children and found to be at risk for conduct disorder. They were assigned randomly (by school) to intervention or control conditions and were followed for 12 years. Intervention lasted 10 years and included parent training, child social-cognitive skills training, reading tutoring, peer-relations enhancement, and classroom curricula and management. Service use was assessed through annual interviews of parents and youth. RESULTS: Youth assigned to preventive intervention had significantly reduced use of professional general health, pediatric, and emergency department services relative to control youth on the basis of parent-report data. For control-group youth, the odds of greater use of general health services for any reason and general health services use for mental health purposes were roughly 30% higher and 56% higher, respectively. On the basis of self-report data, the intervention reduced the likelihood of outpatient mental health services among older adolescents for whom odds of services use were more than 90% higher among control group youth. No differences were found between intervention and control youth on the use of inpatient mental health services. Statistical models controlled for key study characteristics, and potential moderation of the intervention effect was assessed. CONCLUSIONS: Random assignment to the Fast Track prevention program is associated with reduced use of general health and outpatient mental health services in adolescents. Future studies should examine the mechanism of this impact and service use patterns as subjects reach young adulthood. PMID- 20008423 TI - Motavizumab for prophylaxis of respiratory syncytial virus in high-risk children: a noninferiority trial. AB - OBJECTIVE: Palivizumab reduces respiratory syncytial virus (RSV) hospitalization in children at high risk by approximately 50% compared with placebo. We compared the efficacy and safety of motavizumab, an investigational monoclonal antibody with enhanced anti-RSV activity in preclinical studies, with palivizumab. METHODS: This randomized, double-blind, multinational, phase 3, noninferiority trial assessed safety and RSV hospitalization in 6635 preterm infants aged or= 35 weeks and birth weight >or= 2000 g). All measurements were performed with the BiliCheck bilirubinometer, at designated times from 12 to 120 postnatal hours. TcB percentiles for each designated time were calculated and used for the development of an hour-specific nomogram. TcB percentiles for neonates who required phototherapy are also presented. RESULTS: The developed TcB nomogram reflects the natural history of TcB levels in healthy neonates up to the fifth postnatal day. A different pattern of TcB increasing rate was noted in neonates who did and did not require phototherapy but with substantial overlap of TcB values between the 2 groups. CONCLUSIONS: We provide data on TcB levels for the first 120 postnatal hours from a large population of white, healthy, term and near-term neonates. We also present a percentile-based TcB nomogram designated for noninvasive and hour-specific evaluation of neonatal hyperbilirubinemia. PMID- 20008431 TI - Pediatric specialists' beliefs about gastroesophageal reflux disease in premature infants. AB - BACKGROUND: Wide variation exists in the treatment of suspected gastroesophageal reflux disease (GERD) in premature infants; it is unknown to what degree diagnosis and treatment are affected by the treating physician's medical specialty or interpretation of the medical literature. METHODS: This study involved an online survey of board-certified neonatologists, pediatric pulmonologists, and pediatric gastroenterologists about their beliefs regarding the symptoms, diagnosis, and treatment of GERD in premature infants in the NICU on the basis of both clinical impression and interpretation of the literature. RESULTS: A total of 1021 neonatologists, 232 pediatric pulmonologists, and 222 pediatric gastroenterologists participated in the study (47.5% response rate). There was disagreement among specialists in nearly all aspects of the survey. Pulmonologists were most likely to report that respiratory symptoms are caused by GERD (P < .001). Neonatologists were least likely to report that a therapeutic trial of pharmacologic agents would be useful for diagnosing GERD (P < .001) or that lansoprazole, ranitidine, or cimetidine are safe or effective (P < .001). No pharmacologic therapy had >50% of respondents supporting its effectiveness. There was moderate correlation between physician belief based on the medical literature and belief based on clinical impression (Spearman rank correlation: 0.47-0.75). For therapies supported by multiple meta-analyses in infants versus therapies with few infant trials, physicians rated the evidence for effectiveness similarly. CONCLUSIONS: There is wide variation among pediatric specialists regarding beliefs about GERD in premature infants, as well as about the weight of evidence in the medical literature for this patient population. Physician beliefs do not seem to be driven by the degree of evidence in the neonatal literature. With no agreed-on standard of care in the setting of widespread use of antireflux medications, greater understanding is needed about the ways physicians form clinical impressions, access and process medical evidence, and apply it to patient care. PMID- 20008430 TI - Predicting time to hospital discharge for extremely preterm infants. AB - BACKGROUND: As extremely preterm infant mortality rates have decreased, concerns regarding resource use have intensified. Accurate models for predicting time to hospital discharge could aid in resource planning, family counseling, and stimulate quality-improvement initiatives. OBJECTIVES: To develop, validate, and compare several models for predicting the time to hospital discharge for infants <27 weeks' estimated gestational age, on the basis of time-dependent covariates as well as the presence of 5 key risk factors as predictors. PATIENTS AND METHODS: We conducted a retrospective analysis of infants <27 weeks' estimated gestational age who were born between July 2002 and December 2005 and survived to discharge from a Eunice Kennedy Shriver National Institute of Child Health and Human Development Neonatal Research Network site. Time to discharge was modeled as continuous (postmenstrual age at discharge) and categorical (early and late discharge) variables. Three linear and logistic regression models with time dependent covariate inclusion were developed (perinatal factors only, perinatal + early-neonatal factors, and perinatal + early-neonatal + later factors). Models for early and late discharge that used the cumulative presence of 5 key risk factors as predictors were also evaluated. Predictive capabilities were compared by using the coefficient of determination (R(2)) for the linear models and the area under the curve (AUC) of the receiver operating characteristic curve for the logistic models. RESULTS: Data from 2254 infants were included. Prediction of postmenstrual age at discharge was poor. However, models that incorporated later clinical characteristics were more accurate in predicting early or late discharge (AUC: 0.76-0.83 [full models] vs 0.56-0.69 [perinatal factor models]). In simplified key-risk-factors models, the predicted probabilities for early and late discharge compared favorably with the observed rates. Furthermore, the AUC (0.75-0.77) was similar to those of the models that included the full factor set. CONCLUSIONS: Prediction of early or late discharge is poor if only perinatal factors are considered, but it improves substantially with knowledge of later occurring morbidities. Predictive models that use a few key risk factors are comparable to the full models and may offer a clinically applicable strategy. PMID- 20008432 TI - Biological and environmental predictors of behavioral sequelae in children born preterm. AB - OBJECTIVE: By using behavioral outcome measures of children who were born preterm, we evaluated differences between children who were born at term and children who were born at extremely low (ELBW; <1000 g) and very low birth weights (VLBW; 1000-1499 g) and assessed the relationship of birth weight, socioeconomic status, and cognitive ability to behavioral outcome. METHODS: We studied a total of 104 children (aged 7-16 years). Of these, 49 had a preterm birth (31 of ELBW and 18 of VLBW). The remaining 55 were healthy control subjects. Children were administered tests of cognitive ability. Parents and teachers completed behavioral assessments. Multivariate analyses of covariance assessed differences between children who were born at term and those who were born of ELBW and of VLBW on behavioral measures. Hierarchical linear regressions were used to assess relationships among biological (birth weight), environmental (socioeconomic status), intellectual, and behavioral variables. RESULTS: Children who were born at term had fewer parent reports of hyperactivity/inattention and depression/anxiety symptoms than children of ELBW and VLBW. Teacher ratings were not significant between groups. Birth weight was consistently the strongest predictor of parent ratings of behavioral outcome, and intelligence level did not seem to mediate this relationship. CONCLUSIONS: Negative behavioral sequelae of preterm birth remain significant in middle childhood and adolescence, although the contribution of multiple factors to neurobehavioral outcome is complex. Research to assess these relationships, integrated with anatomic and functional neuroimaging, is needed to advance knowledge and improve outcomes for children who are born preterm. PMID- 20008433 TI - Part-time work among pediatricians expands. AB - OBJECTIVE: The objective of this study was to track trends in part-time employment among pediatricians from 2000 to 2006 and to examine differences within subgroups of pediatricians. METHODS: As part of the Periodic Survey of Fellows, national random samples of American Academy of Pediatrics members were surveyed in 2000, 2003, and 2006. These surveys shared questions concerning working part-time and other practice characteristics. Roughly 1600 pediatricians were included in each random sample. Totals of 812 (51%), 1020 (63%), and 1013 (62%) pediatricians completed the surveys in 2000, 2003, and 2006, respectively. Analyses were limited to nonretired, posttrainee pediatricians. RESULTS: The number of pediatricians who reported that they work part-time increased from 15% in 2000, to 20% in 2003, to 23% in 2006. The pattern of increased part-time work from 2000 to 2006 held for many subgroups, including men, women, pediatricians who were younger than 40 years, pediatricians who were aged >or=50 years, pediatricians who worked in an urban inner city, pediatricians who worked in suburban areas, general pediatricians, and subspecialist pediatricians. Those who were working part-time were more satisfied within their professional and personal activities. Part-time pediatricians worked on average 14.3 fewer hours per week in direct patient care. CONCLUSIONS: Increases in part-time work are apparent throughout pediatrics. The possible continued growth of part-time is an important trend within the field of pediatrics that will need to be monitored. PMID- 20008435 TI - Patent medicines and secret remedies. PMID- 20008436 TI - Women challenge Irish law on abortion in Europe on human rights grounds. PMID- 20008440 TI - From flower graves to breast clinics. PMID- 20008439 TI - Mechanisms of epigenetic silencing of the Rassf1a gene during estrogen-induced breast carcinogenesis in ACI rats. AB - Breast cancer, the most common malignancy in women, emerges through a multistep process, encompassing the progressive sequential evolution of morphologically distinct stages from a normal cell to hyperplasia (with and without atypia), carcinoma in situ, invasive carcinoma and metastasis. The success of treatment of breast cancer could be greatly improved by the detection at early stages of cancer. In the present study, we investigated the underlying molecular mechanisms involved in breast carcinogenesis in Augustus and Copenhagen-Irish female rats, a cross between the ACI strains, induced by continuous exposure to 17beta estradiol. The results of our study demonstrate that early stages of estrogen induced breast carcinogenesis are characterized by altered global DNA methylation, aberrant expression of proteins responsible for the proper maintenance of DNA methylation pattern and epigenetic silencing of the critical Rassf1a (Ras-association domain family 1, isoform A) tumor suppressor gene. Interestingly, transcriptional repression of the Rassf1a gene in mammary glands during early stages of breast carcinogenesis was associated with an increase in trimethylation of histones H3 lysine 9 and H3 lysine 27 and de novo CpG island methylation and at the Rassf1a promoter and first exon. In conclusion, we demonstrate that epigenetic alterations precede formation of preneoplastic lesions indicating the significance of epigenetic events in induction of oncogenic pathways in early stages of carcinogenesis. PMID- 20008441 TI - Animated ophthalmology. PMID- 20008442 TI - Selling patients. PMID- 20008444 TI - Induction of autoimmunity in brown Norway rats by oral and parenteral administration of nickel chloride. AB - Nickel intolerance owing to sensitization is a growing problem. The main objective of this study was to examine the relationship between nickel chloride and induction of autoimmunity in genetically susceptible rats. Thirty brown Norway rats were randomized into four treatment groups; the first and second groups received nickel chloride 4.5 mg in 0.2 ml normal saline either orally or subcutaneously, and the third and fourth groups (controls) received normal saline (0.9%) 0.2 ml through the same routes. A significant number of rats (P < 0.05) that received nickel chloride by the subcutaneous or oral routes showed a high level of serum antinuclear antibody (ANA) compared with controls. A significant number of rats (P < 0.05) that received nickel chloride by the subcutaneous route showed high serum anti-SSA, but the number of rats with anti-SSA was insignificant in the group that received nickel by the oral route. Other autoantibodies found in both groups (anti-double-stranded (ds)DNA, anti-Smith, anti-SSB) showed a gradual increase, but the number of rats with positive titers post exposure was not significant statistically. Nickel chloride exposure in the rats appeared to induce the development of autoimmunity. A longer duration following exposure to nickel chloride seems to be associated with greater risks. PMID- 20008445 TI - Frequency of neuro-psychiatric dysfunction in anti-SSA/SSB exposed children with and without neonatal lupus. AB - Neonatal lupus is a model of passively acquired autoimmunity whereby anti-SSA/Ro SSB/La antibodies target the fetal heart and neonatal skin in a minority of cases. Since neuro-psychiatric impairment has been reported in humans and mice exposed prenatally to a variety of maternal autoantibodies including anti-Ro/La, this study was initiated to evaluate the potential neurotoxic effects of these specific autoantibodies and the overall frequency of autoimmune diseases, general health, and somatic growth of children with neonatal lupus and their unaffected siblings. In addition to the general health questionnaires maintained on family members enrolled in the Research Registry for Neonatal Lupus (RRNL), specific questionnaires related to neuro-psychiatric development were sent to all mothers whose children (both affected and unaffected) were older than 5 years of age. Controls consisted of healthy friends. Of 121 anti-Ro exposed children meeting the inclusion criteria, information was returned on 104 (33 cardiac manifestations of neonatal lupus, 20 rash, and 51 unaffected siblings) and 22 of the friend controls. The mean age of all of the children was 14.5 years (range 5 39). In total, 42 (40%) of the 104 anti-Ro exposed children were reported to have a neuro-psychiatric disorder, compared with 6 (27%) of the friend controls (p = 0.34). For 8 (24%) of the congenital heart block (CHB) children (6 boys, 2 girls) the mothers reported attention problems. Four, all boys, were on stimulants. Of the rash children, 4 (20%) (2 boys, 2 girls) had attention problems with one boy on Ritalin. Of the unaffected siblings, 9 (18%) (8 boys and 1 girl) had attention problems with 3 boys on stimulants. One (5%) of the control children (a girl) had attention problems, not requiring therapy. There was no statistical difference in attention problems between the groups (p = 0.120). Behavioral problems were present in all groups with no statistical differences noted. The prevalence of depression, anxiety, developmental delays, learning, hearing, and speech problems were not significantly different between groups. In the CHB children, one boy has nephrotic syndrome and one girl has psoriasis. In the rash children, one girl has juvenile rheumatoid arthritis. In the unaffected group there are five children with autoimmune diseases, two with inflammatory bowel diseases (one boy and one girl), one boy has a spondyloarthropathy, one girl has alopecia areata and one young woman has Antiphospholipid syndrome. In the control group one boy has Henoch Schonlein purpura. There were four cases of hypothyroidism, possibly secondary to Hashimoto's thyroiditis, three in boys with CHB and one in a girl with rash. None of the unaffected siblings or controls had hypothyroidism. Parental reporting of neuro-psychiatric abnormalities was high in anti-Ro exposed children regardless of the neonatal lupus manifestation. However, medication use was limited and although the frequency of this reporting was greater than friend controls, it did not reach significance. PMID- 20008446 TI - Evaluation of akathisia in patients with schizophrenia, schizoaffective disorder, or bipolar I disorder: a post hoc analysis of pooled data from short- and long term aripiprazole trials. AB - The objective of this article is to assess the clinical characteristics of akathisia in patients with schizophrenia, schizoaffective disorder, or bipolar I disorder receiving aripiprazole, haloperidol, olanzapine, or placebo. We conducted post hoc analyses of pooled safety data from trials in patients with schizophrenia, schizoaffective disorder, and bipolar I disorder. Outcome measures included the incidence of akathisia, time to onset, duration, severity, and discontinuation due to akathisia, concomitant use of benzodiazepines and/or anticholinergics, Barnes Akathisia Rating Scale (BARS) scores, and the correlation between antipsychotic efficacy and akathisia. The results for schizophrenia and schizoaffective disorder were as follows: akathisia in 9% of aripiprazole- and 6% of placebo-treated patients; 12.5% of aripiprazole- versus 24% of haloperidol-treated patients; 11% of aripiprazole- versus 6% of olanzapine treated patients. Bipolar I disorder: akathisia in 18% of aripiprazole- and 5% of placebo-treated patients. The clinical characteristics of akathisia were similar between each data set, regardless of disease. Akathisia was generally mild-to moderate in severity. Discontinuation due to akathisia was low in both the schizophrenia trials (aripiprazole 0.3%; placebo 0%; aripiprazole 0.9%; haloperidol 2.3%; aripiprazole 1.2%; olanzapine 0.2%) and the bipolar trials (aripiprazole 2.3%; placebo 0%). Treatment-emergent akathisia was not associated with a poorer clinical response. In conclusion, akathisia with aripiprazole occurred early in treatment, was mild-to-moderate in severity, led to few study discontinuations, and did not compromise therapeutic efficacy. PMID- 20008447 TI - Emergence of clonally related Klebsiella pneumoniae isolates of sequence type 258 producing KPC-2 carbapenemase in Belgium. PMID- 20008448 TI - Characterization of fluoroquinolone resistance in a clinical isolate of Pseudomonas stutzeri. PMID- 20008449 TI - Reducing antimicrobial resistance in the community by restricting prescribing: can it be done? AB - The strategy of decreasing antimicrobial prescribing to reduce existing antimicrobial resistance appears attractive, but its effectiveness, particularly in the community setting, remains unclear. Contrasting results obtained from the relatively few studies in this area confuse matters further. Prescribing reductions have successfully reduced the prevalence of resistance among respiratory pathogens, but in these cases single bacterial clones dominated the resistant population. In contrast, this strategy has not succeeded in reducing plasmid-encoded resistance among Escherichia coli. The reasons why some prescribing restriction policies are more successful than others are complex, with the three key determinants being the fitness cost of resistance, the clonal structure of the resistant bacterial population and co-selection of resistant organisms by other antimicrobials. The resistant bacterial phenotypes that are likely to be the easiest to eliminate will be those composed of relatively clonal populations that bear a fitness cost of resistance and are not significantly subjected to co-selection by other antimicrobials. Plasmid-encoded resistance seldom meets these criteria and, hence, is likely to be the most difficult to reduce. PMID- 20008450 TI - Red blood cell distribution width and risk of cardiovascular events and mortality in a community cohort in Taiwan. AB - The authors investigated whether red blood cell distribution width (RDW) was associated with the development of cardiovascular disease (CVD) events and mortality in a community cohort in Taiwan. The influence of anemia on the association was also assessed. RDW levels were measured in 3,226 participants aged 35 years or older who reported no CVD or cancer at baseline in 1990. During a median follow-up period of 15.9 years (1990-2007), 358 participants experienced stroke and/or coronary heart disease, and 810 participants died. The multivariate adjusted hazard ratio for subjects in the highest RDW quartile as compared with the lowest quartile was 1.46 for both all-cause mortality (95% confidence interval: 1.17, 1.81) and non-CVD mortality (95% confidence interval: 1.13, 1.88) (P for trend < 0.01 for both) but was not significant for CVD morbidity and mortality. Further analyses showed that in comparison with participants with low RDW and no anemia, persons with high RDW but no anemia had elevated risks of all cause mortality and non-CVD mortality. The authors conclude that elevated RDW values are associated with increased risk of mortality but not the development of CVD in the general population. RDW may precede anemia in predicting the risk of non-CVD death. PMID- 20008452 TI - Pleiotropic regulatory locus 1 (PRL1) integrates the regulation of sugar responses with isoprenoid metabolism in Arabidopsis. AB - The biosynthesis of isoprenoids in plant cells occurs from precursors produced in the cytosol by the mevalonate (MVA) pathway and in the plastid by the methylerythritol 4-phosphate (MEP) pathway, but little is known about the mechanisms coordinating both pathways. Evidence of the importance of sugar signaling for such coordination in Arabidopsis thaliana is provided here by the characterization of a mutant showing an increased accumulation of MEP-derived isoprenoid products (chlorophylls and carotenoids) without changes in the levels of relevant MEP pathway transcripts, proteins, or enzyme activities. This mutant was found to be a new loss-of-function allele of PRL1 (Pleiotropic Regulatory Locus 1), a gene encoding a conserved WD-protein that functions as a global regulator of sugar, stress, and hormone responses, in part by inhibition of SNF1 related protein kinases (SnRK1). Consistent with the reported role of SnRK1 kinases in the phosphorylation and inactivation of the main regulatory enzyme of the MVA pathway (hydroxymethylglutaryl coenzyme-A reductase), its activity but not transcript or protein levels was reduced in prl1 seedlings. However, the accumulation of MVA-derived end products (sterols) was unaltered in mutant seedlings. Sucrose supplementation to wild-type seedlings phenocopied the prl1 mutation in terms of isoprenoid metabolism, suggesting that the observed isoprenoid phenotypes result from the increased sugar accumulation in the prl1 mutant. In summary, PRL1 appears to coordinate isoprenoid metabolism with sugar, hormone, and stress responses. PMID- 20008451 TI - Metabolomic, transcriptional, hormonal, and signaling cross-talk in superroot2. AB - Auxin homeostasis is pivotal for normal plant growth and development. The superroot2 (sur2) mutant was initially isolated in a forward genetic screen for auxin overproducers, and SUR2 was suggested to control auxin conjugation and thereby regulate auxin homeostasis. However, the phenotype was not uniform and could not be described as a pure high auxin phenotype, indicating that knockout of CYP83B1 has multiple effects. Subsequently, SUR2 was identified as CYP83B1, a cytochrome P450 positioned at the metabolic branch point between auxin and indole glucosinolate metabolism. To investigate concomitant global alterations triggered by knockout of CYP83B1 and the countermeasures chosen by the mutant to cope with hormonal and metabolic imbalances, 10-day-old mutant seedlings were characterized with respect to their transcriptome and metabolome profiles. Here, we report a global analysis of the sur2 mutant by the use of a combined transcriptomic and metabolomic approach revealing pronounced effects on several metabolic grids including the intersection between secondary metabolism, cell wall turnover, hormone metabolism, and stress responses. Metabolic and transcriptional cross talks in sur2 were found to be regulated by complex interactions between both positively and negatively acting transcription factors. The complex phenotype of sur2 may thus not only be assigned to elevated levels of auxin, but also to ethylene and abscisic acid responses as well as drought responses in the absence of a water deficiency. The delicate balance between these signals explains why minute changes in growth conditions may result in the non-uniform phenotype. The large phenotypic variation observed between and within the different surveys may be reconciled by the complex and intricate hormonal balances in sur2 seedlings decoded in this study. PMID- 20008453 TI - Neural dynamics of the intention to speak. AB - When we talk we communicate our intentions. Although the origin of intentional action is debated in cognitive neuroscience, the question of how the brain generates the intention in speech remains still open. Using magnetoencephalography, we investigated the cortical dynamics engaged when healthy subjects attended to either their intention to speak or their actual speech. We found that activity in the right and left parietal cortex increased before subjects became aware of intending to speak. Within the time window of parietal activation, we also observed a transient left frontal activity in Broca's area, a crucial region for inner speech. During attention to speech, neural activity was detected in left prefrontal and temporal areas and in the temporoparietal junction. In agreement with previous results, our findings suggest that the parietal cortex plays a multimodal role in monitoring intentional mechanisms in both action and language. The coactivation of parietal regions and Broca's area may constitute the cortical circuit specific for controlling intentional processes during speech. PMID- 20008454 TI - Acute effects of complexity in aroma composition on satiation and food intake. AB - The acute effect of complexity in aroma composition on perceived satiation and food intake was investigated in 41 young, healthy, and normal weight subjects. Subjects consumed 2 different strawberry-aromatized sweetened yogurt products (i.e., test and placebo product) in either an olfactometer-aided or an ad libitum eating experimental design. The test product was aromatized with a multicomponent strawberry aroma, whereas the placebo product was aromatized with a single component strawberry aroma. Compared to placebo, subjects felt significantly more satiated during aroma stimulation with the multicomponent strawberry aroma in the olfactometer-aided setting. Additionally, perceived satiation was significantly increased 10-15 min after consumption of the multicomponent strawberry-aromatized sweetened yogurt product in the ad libitum eating setting. There was no effect on the amount of strawberry-aromatized sweetened yogurt product consumed ad libitum. Apart from the differences in timing of the appetite-regulating effects, both experimental settings demonstrated that the multicomponent strawberry aroma, which was perceived as being more complex, yet of similar aroma quality, intensity, and pleasantness compared with the single-component strawberry aroma, was able to enhance perceived satiation. The methodology of the olfactometer aided aroma stimulation proved to be representative of a real-life setting with regard to aroma exposure and satiation. Food products, which are perceived as being more complex, have been suggested to delay the development of sensory satiation as a result of implicitly cueing for variation. The present results may be explained by increased sensory stimulation, due to concurrent exposure to multiple aroma components cueing for sensorily similar strawberry perception. PMID- 20008455 TI - Intellectual enrichment is linked to cerebral efficiency in multiple sclerosis: functional magnetic resonance imaging evidence for cognitive reserve. AB - The cognitive reserve hypothesis helps to explain the incomplete relationship between brain disease and cognitive status in people with neurologic diseases, including Alzheimer's; disease and multiple sclerosis. Lifetime intellectual enrichment (estimated with education or vocabulary knowledge) lessens the negative impact of brain disease on cognition, such that people with greater enrichment are able to withstand more severe neuropathology before suffering cognitive impairment or dementia. The current research is the first to investigate directly the relationship between intellectual enrichment and an index of cerebral activity (the blood oxygen level dependent signal) in a neurologic sample. Multiple sclerosis patients completed a vocabulary-based estimate of lifetime intellectual enrichment. Disease severity was estimated with brain atrophy. Cognitive status was measured with the Symbol Digit Modalities Test. Cerebral activity (functional magnetic resonance imaging blood oxygen level dependent signal) and behavioural performance (accuracy, reaction time) were recorded during the visual N-Back working memory task (three levels of demand: 0 , 1-, 2-Back). All patients produced perfect/nearly perfect accuracy during lower demands (0- and 1-Back), and reaction time was unrelated to intellectual enrichment; however, voxelwise partial correlations controlling for brain atrophy revealed strong positive correlations between intellectual enrichment and cerebral activity within the brain's; default network (e.g. anterior and posterior cingulate corticies), indicating that patients with greater enrichment were able to maintain resting state activity during cognitive processing better. In turn, intellectual enrichment was negatively associated with prefrontal recruitment, suggesting that patients with lesser enrichment required more cerebral resources to perform the same cognitive task as patients with greater enrichment. This same pattern of enrichment-related cerebral activity was observed when cognitive demands increased (2-Back), and intellectual enrichment was negatively associated with reaction time. Principle components analysis revealed a single cognitive reserve network across tasks (greater default network, lesser prefrontal recruitment). Expression of this network almost fully mediated the positive relationship between intellectual enrichment and cognitive status (Symbol Digit Modalities Test). Also, expression of this network was positively associated with brain atrophy when controlling for cognitive status, indicating that patients with greater expression of this network can withstand more severe brain disease before exhibiting cognition similar to patients with lesser network expression. Of note, similar functional magnetic resonance imaging research with healthy adults has not found an association between intelligence and cerebral efficiency. The unique relationship between intellectual enrichment and cerebral efficiency in neurologic patients is consistent with the cognitive reserve hypothesis, which does not posit that enrichment leads to gains in neurocognitive functioning per se; rather, enrichment protects against neurocognitive decline secondarily to disease. PMID- 20008456 TI - Population pharmacokinetics of 3,4-methylenedioxymethamphetamine and main metabolites in rats. AB - The pharmacokinetics of the recreational drug 3,4-methylenedioxymethamphetamine (MDMA) and its mains metabolites have never been modeled together. We therefore designed a model with which to analyze the pharmacokinetics of MDMA, 3,4 methylenedioxyamphetamine (MDA), 4-hydroxy-3-methoxymethamphetamine (HMMA), and 4 hydroxy-3-methoxyamphetamine (HMA) and to test the effect of covariates like gender and body weight on the pharmacokinetics. Rats (18 males and 18 females) were given 1 mg/kg MDMA iv, and the concentrations of MDMA, MDA, and HMMA were measured by high-performance liquid chromatography-mass spectrometry. Another 30 rats (15 males) were given 1 mg/kg MDA, and MDA and HMA were measured. A population pharmacokinetic model was developed to describe the changes in MDMA, HMMA, MDA, and HMA concentrations over time and to estimate interanimal variability. The influence of gender was tested using a likelihood ratio test. Estimated exposures of males and females to MDMA and its metabolites were compared using the Wilcoxon nonparametric test. An integrated six-compartment model adequately described the data. MDMA (two compartments) was transformed irreversible to HMMA (one compartment) and MDA (two compartments), which then produced HMA (one compartment). All rate constants were first order. Females given MDMA had significantly smaller MDMA distribution volumes than males, and they converted less MDMA to MDA than did males. Our MDMA, MDA, HMA, and HMMA model is suitable for examining the relationship between drug concentrations and its pharmacological/toxicological effects. Male rats were exposed to significantly more MDA and HMA than were females, which could explain why males are more sensitive to MDMA toxic effects than females. PMID- 20008458 TI - Evolution of relative reading frame bias in unidirectional prokaryotic gene overlaps. AB - Pairs of unidirectional (same strand) genes can overlap in one of two phases (relative reading frames). There is a striking bias in the relative abundance of prokaryotic gene overlaps in the two possible phases. A simple model is presented based on unidirectional gene overlaps evolving from nonoverlapping gene pairs, through the adoption of alternative start codons by the downstream genes. Potential alternative start codons within upstream gene sequences were found to occur at greater frequencies in one phase, corresponding to the most prevalent phase of gene overlaps. We therefore suggest that the phase bias of overlapping genes is primarily a consequence of the N-terminal extension of downstream genes through adoption of new start codons. PMID- 20008457 TI - Recombination yet inefficient selection along the Drosophila melanogaster subgroup's fourth chromosome. AB - A central goal of evolutionary genetics is an understanding of the forces responsible for the observed variation, both within and between species. Theoretical and empirical work have demonstrated that genetic recombination contributes to this variation by breaking down linkage between nucleotide sites, thus allowing them to behave independently and for selective forces to act efficiently on them. The Drosophila fourth chromosome, which is believed to experience no-or very low-rates of recombination has been an important model for investigating these effects. Despite previous efforts, central questions regarding the extent of recombination and the predominant modes of selection acting on it remain open. In order to more comprehensively test hypotheses regarding recombination and its potential influence on selection along the fourth chromosome, we have resequenced regions from most of its genes from Drosophila melanogaster, D. simulans, and D. yakuba. These data, along with available outgroup sequence, demonstrate that recombination is low but significantly greater than zero for the three species. Despite there being recombination, there is strong evidence that its frequency is low enough to have rendered selection relatively inefficient. The signatures of relaxed constraint can be detected at both the level of polymorphism and divergence. PMID- 20008459 TI - GPR56 as a novel marker identifying the CD56dull CD16+ NK cell subset both in blood stream and in inflamed peripheral tissues. AB - To define novel human NK cell markers, we generated two mAbs specific for G protein-coupled receptor 56 (GPR56), a surface glycoprotein that appears to be involved in cell-to-cell and cell-to-matrix interactions. GPR56 has been described in selected normal tissues, and in certain tumors, while, as yet, its expression on leukocytes is unknown. In this study, we show that anti-GPR56 mAbs, among leukocytes, prevalently recognize NK cells. In particular, these mAbs brightly stain CD56(dull) CD16(+) NK cells while react poorly with CD56(bright) CD16(+/-) NK cells. Consistently, we found that GPR56 was expressed on NK cells populating inflamed peripheral tissues while it was absent in lymph node-derived NK cells. We also show that activating stimuli, such as cytokines or exposure to monocyte-derived dendritic cell, down-regulate NK cell expression of GPR56 both at the protein and at the transcriptional level. Interestingly, IL-18, known to induce de novo expression of CCR7 on CD56(dull) CD16(+) NK cells, displayed the highest capability of modulating GPR56. Thus, together with the identification of GPR56 as a novel marker capable of discriminating different NK cells subsets, our data suggest that GPR56 may take part to the mechanisms regulating NK cell migration through the blood stream, peripheral tissues and lymph nodes. PMID- 20008460 TI - Antioxidant Activity of Lawsonia inermis Extracts Inhibits Chromium(VI)-Induced Cellular and DNA Toxicity. AB - Hexavalent chromium Cr(VI) is a very strong oxidant which consequently causes high cytotoxicity through oxidative stress. Prevention of Cr(VI)-induced cellular damage has been sought in this study in aqueous and methanolic extracts of Lawsonia inermis Linn. (Lythraceae), commonly known as Henna. The extracts showed significant (P < .05) potential in scavenging free radicals (DPPH(*) and ABTS(*+)) and Fe(3+), and in inhibiting lipid peroxidation. DNA damage caused by exposure of pBR322 to Cr(VI)-UV is markedly inhibited by both extracts in varying degrees. A distinct decline in Cr(VI)-induced cytotoxicity was noticed in MDA-MB 435S (human breast carcinoma) cells with an increase in dosage of both extracts individually. Furthermore, both extracts proved to contain a high content of phenolic compounds which were found to have a strong and significant (P < .05) positive correlation to the radical scavenging potential, lipid peroxidation inhibition capacity and cyto-protective efficiency against Cr(VI)-induced oxidative cellular damage. HPLC analysis identified some of the major phenolic compounds in both extracts, which might be responsible for the antioxidant potential and the properties of DNA and cyto-protection. This study contributes to the search for natural resources that might yield potent therapeutic drugs against Cr(VI)-induced oxidative cell damage. PMID- 20008461 TI - Expression of the ethylene biosynthetic machinery in maize roots is regulated in response to hypoxia. AB - Ethylene regulates plant growth in response to many adverse environmental conditions, including the induction of aerenchyma, i.e. the formation of air spaces, in flooded roots in an effort to maintain oxygen levels. In this work, quantitative RT-PCR and in situ RNA hybridization were used to determine how the expression of the ethylene biosynthetic machinery in maize roots is spatially and temporally regulated following exposure to 4% oxygen (i.e. hypoxia) for up to 24 h, conditions that induced aerenchyma formation in the fully-expanded region of the root and reduced cytoplasmic density throughout the root. Expression of ACC oxidase, the ethylene forming enzyme, was observed in the root cap, protophloem sieve elements, and companion cells associated with metaphloem sieve elements. Exposure to 4% oxygen induced ACC oxidase expression in these cell types as well as in the root cortex. ACC synthase, which generates the ethylene precursor, was expressed in the root cap and the cortex and its expression was induced in cortical cells following low oxygen treatment. The induction of expression of the ethylene biosynthetic machinery was accompanied by an induction of ethylene evolution and a reduced rate of root growth. These results suggest that maize roots respond to conditions of hypoxia by inducing the spatially restricted expression of the ethylene biosynthetic machinery, resulting in increased ethylene production. PMID- 20008462 TI - Genetic evidence that two independent S-loci control RNase-based self incompatibility in diploid strawberry. AB - The self-incompatibility mechanism that reduces inbreeding in many plants of the Rosaceae is attributed to a multi-allelic S locus which, in the Prunoideae and Maloideae subfamilies, comprises two complementary genes, a stylar-expressed S RNase and a pollen-expressed SFB. To elucidate incompatibility in the subfamily Rosoideae, stylar-specific RNases and self-(in)compatibility status were analysed in various diploid strawberries, especially Fragaria nubicola and F. viridis, both self-incompatible, and F. vesca, self-compatible, and in various progenies derived from them. Unexpectedly, two unlinked RNase loci, S and T, were found, encoding peptides distinct from Prunoideae and Maloideae S-RNases; the presence of a single active allele at either is sufficient to confer self-incompatibility. By contrast, in diploid Maloideae and Prunoideae a single locus encodes S-RNases that share several conserved regions and two active alleles are required for self incompatibility. Our evidence implicates the S locus in unilateral inter-specific incompatibility and shows that S and T RNases can, remarkably, confer not only allele-specific rejection of cognate pollen but also unspecific rejection of Sn Tn pollen, where n indicates a null allele, consistent with the the presence of the pollen component, SFB, activating the cognitive function of these RNases. Comparison of relevant linkage groups between Fragaria and Prunus suggests that Prunus S-RNases, unique in having two introns, may have resulted from gene conversion in an ancestor of Prunus. In addition, it is shown that there is a non S locus that is essential for self-incompatibility in diploid Fragaria. PMID- 20008463 TI - Cones perform a non-linear transformation on natural stimuli. AB - Visual information in natural scenes is distributed over a broad range of intensities and contrasts. This distribution has to be compressed in the retina to match the dynamic range of retinal neurons. In this study we examined how cones perform this compression and investigated which physiological processes contribute to this operation. M- and L-cones of the goldfish were stimulated with a natural time series of intensities (NTSI) and their responses were recorded. The NTSI displays an intensity distribution which is skewed towards the lower intensities and has a long tail into the high intensity region. Cones transform this skewed distribution into a more symmetrical one. The voltage responses of the goldfish cones were compared to those of a linear filter and a non-linear biophysical model of the photoreceptor. The results show that the linear filter under-represents contrasts at low intensities compared to the actual cone whereas the non-linear biophysical model performs well over the whole intensity range used. Quantitative analysis of the two approaches indicates that the non-linear biophysical model can capture 91 +/- 5% of the coherence rate (a biased measure of information rate) of the actual cone, where the linear filter only reaches 48 +/- 8%. These results demonstrate that cone photoreceptors transform an NTSI in a non-linear fashion. The comparison between current clamp and voltage clamp recordings and analysis of the behaviour of the biophysical model indicates that both the calcium feedback loop in the outer segment and the hydrolysis of cGMP are the major components that introduce the specific non-linear response properties found in the goldfish cones. PMID- 20008464 TI - Electrophysiological identification of glucose-sensing neurons in rat nodose ganglia. AB - The vagal afferent system is strategically positioned to mediate rapid changes in motility and satiety in response to systemic glucose levels. In the present study we aimed to identify glucose-excited and glucose-inhibited neurons in nodose ganglia and characterize their glucose-sensing properties. Whole-cell patch-clamp recordings in vagal afferent neurons isolated from rat nodose ganglia demonstrated that 31/118 (26%) neurons were depolarized after increasing extracellular glucose from 5 to 15 mm; 19/118 (16%) were hyperpolarized, and 68/118 were non-responsive. A higher incidence of excitatory response to glucose occurred in gastric- than in portal vein-projecting neurons, the latter having a higher incidence of inhibitory response. In glucose-excited neurons, elevated glucose evoked membrane depolarization (11 mV) and an increase in membrane input resistance (361 to 437 M). Current reversed at 99 mV. In glucose-inhibited neurons, membrane hyperpolarization (13 mV) was associated with decreased membrane input resistance (383 to 293 M). Current reversed at 97 mV. Superfusion of tolbutamide, a K(ATP) channel sulfonylurea receptor blocker, elicited identical glucose-excitatory but not glucose-inhibitory responses. Kir6.2 shRNA transfection abolished glucose-excited but not glucose-inhibited responses. Phosphatidylinositol bisphosphate (PIP(2)) depletion using wortmannin increased the fraction of glucose-excited neurons from 26% to 80%. These results show that rat nodose ganglia have glucose-excited and glucose-inhibited neurons, differentially distributed among gastric- and portal vein-projecting nodose neurons. In glucose-excited neurons, glucose metabolism leads to K(ATP) channel closure, triggering membrane depolarization, whereas in glucose-inhibited neurons, the inhibitory effect of elevated glucose is mediated by an ATP independent K(+) channel. The results also show that PIP(2) can determine the excitability of glucose-excited neurons. PMID- 20008465 TI - NMDAR-mediated EPSCs are maintained and accelerate in time course during maturation of mouse and rat auditory brainstem in vitro. AB - NMDA receptors (NMDARs) mediate a slow EPSC at excitatory glutamatergic synapses throughout the brain. In many areas the magnitude of the NMDAR-mediated EPSC declines with development and is associated with changes in subunit composition, but the mature channel composition is often unknown. We have employed the calyx of Held terminal with its target, the principal neuron of the medial nucleus of the trapezoid body (MNTB), to examine the NMDAR-mediated EPSC during synapse maturation from P10 to P40. Our data show that the calyx has reached a mature state by around P18. The NMDAR-mediated EPSC amplitude (and dominant decay ) fell from around 5 nA (: 40-50 ms) at P10/11 to 0.3-0.5 nA (: 10-15 ms) by P18. The mature NMDAR-EPSC showed no sensitivity to ifenprodil, indicating lack of NR2B subunits, and no block by submicromolar concentrations of zinc, consistent with NR1-1b subunit expression. Additionally, from P11 to P18 there was a reduction in voltage-dependent block and the apparent dissociation constant for [Mg(2+)](o) (K(o)) changed from 7.5 to 14 mm. Quantitative PCR showed that the relative expression of NR2A and NR2C increased, while immunohistochemistry confirmed the presence of NR2A, NR2B and NR2C protein. Although the mature NMDAR-EPSC is small, it is well coupled to NO signalling, as indicated by DAR-4M imaging. We conclude that native mature NMDAR channels at the calyx of Held have a fast time course and reduced block by [Mg(2+)](o), consistent with dominance of NR2C subunits and functional exclusion of NR2B subunits. The pharmacology suggests a single channel type and we postulate that the mature NMDARs consist of heterotrimers of NR1-1b NR2A-NR2C. PMID- 20008466 TI - Ozone activates airway nerves via the selective stimulation of TRPA1 ion channels. AB - Inhalation of ozone is a major health risk in industrialized nations. Ozone can impair lung function and induce respiratory symptoms through sensory neural mediated pathways, yet the specific interaction of ozone with airway sensory nerves has yet to be elucidated. Here we demonstrate, using a vagally innervated ex vivo tracheal-lung mouse preparation, that ozone selectively and directly evokes action potential discharge in a subset of nociceptive bronchopulmonary nerves, namely slow conducting C-fibres. Sensitivity to ozone correlated with the transient receptor potential (TRP) A1 agonist, cinnamaldehyde, with ozone having no effect on cinnamaldehyde-insensitive fibres. C-fibre responses to ozone were abolished by ruthenium red (TRP inhibitor). Ozone also stimulated a subset of nociceptive sensory neurones isolated from vagal ganglia of wild-type mice, but failed to activate neurones isolated from transient receptor potential ankyrin 1 (TRPA1) knockout mice. Ozone activated HEK293 cells transfected with TRPA1, but failed to activate non-transfected HEK293 or HEK293 transfected with the capsaicin-sensitive transient receptor potential vanilloid 1 (TRPV1) channel. Thus, ozone is not an indiscriminate neuronal activator, but rather it potently and selectively activates a subset of airway C-fibres by directly stimulating TRPA1. PMID- 20008467 TI - Sodium accumulation promotes diastolic dysfunction in end-stage heart failure following Serca2 knockout. AB - Alterations in trans-sarcolemmal and sarcoplasmic reticulum (SR) Ca(2+) fluxes may contribute to impaired cardiomyocyte contraction and relaxation in heart failure. We investigated the mechanisms underlying heart failure progression in mice with conditional, cardiomyocyte-specific excision of the SR Ca(2+)-ATPase (SERCA) gene. At 4 weeks following gene deletion (4-week KO) cardiac function remained near normal values. However, end-stage heart failure developed by 7 weeks (7-week KO) as systolic and diastolic performance declined. Contractions in isolated myocytes were reduced between 4- and 7-week KO, and relaxation was slowed. Ca(2+) transients were similarly altered. Reduction in Ca(2+) transient magnitude resulted from complete loss of SR Ca(2+) release between 4- and 7-week KO, due to loss of a small remaining pool of SERCA2. Declining SR Ca(2+) release was partly offset by increased L-type Ca(2+) current, which was facilitated by AP prolongation in 7-week KO. Ca(2+) entry via reverse-mode Na(+)-Ca(2+) exchange (NCX) was also enhanced. Up-regulation of NCX and plasma membrane Ca(2+)-ATPase increased Ca(2+) extrusion rates in 4-week KO. Diastolic dysfunction in 7-week KO resulted from further SERCA2 loss, but also impaired NCX-mediated Ca(2+) extrusion following Na(+) accumulation. Reduced Na(+)-K(+)-ATPase activity contributed to the Na(+) gain. Normalizing [Na(+)] by dialysis increased the Ca(2+) decline rate in 7-week KO beyond 4-week values. Thus, while SERCA2 loss promotes both systolic and diastolic dysfunction, Na(+) accumulation additionally impairs relaxation in this model. Our observations indicate that if cytosolic Na(+) gain is prevented, up-regulated Ca(2+) extrusion mechanisms can maintain near-normal diastolic function in the absence of SERCA2. PMID- 20008468 TI - Dihydrotestosterone activates the MAPK pathway and modulates maximum isometric force through the EGF receptor in isolated intact mouse skeletal muscle fibres. AB - It is generally believed that steroid hormones have both genomic and non-genomic (rapid) actions. Although the latter form an important component of the physiological response of these hormones, little is known about the cellular signalling pathway(s) mediating these effects and their physiological functions in adult mammalian skeletal muscle fibres. Therefore, the primary aim of this study was to investigate the non-genomic actions of dihydrotestosterone (DHT) and their physiological role in isolated intact mammalian skeletal muscle fibre bundles. Our results show that treating the fibre bundles with physiological concentrations of DHT increases both twitch and tetanic contractions in fast twitch fibres. However, it decreases them in slow twitch fibres. These changes in force are accompanied by an increase in the phosphorylation of MAPK/ERK1/2 in both fibre types and that of regulatory myosin light chains in fast twitch fibres. Both effects were insensitive to inhibitors of Src kinase, androgen receptor, insulin-like growth factor 1 receptor and platelet-derived growth factor receptor. However, they were abolished by the MAPK/ERK1/2 kinase inhibitor PD98059 and the epidermal growth factor (EGF) receptor inhibitor tyrphostin AG 1478. In contrast, testosterone had no effect on force and increased the phosphorylation of ERK1/2 in slow twitch fibres only. From these results we conclude that sex steroids have non-genomic actions in isolated intact mammalian skeletal muscle fibres. These are mediated through the EGF receptor and one of their main physiological functions is the enhancement of force production in fast twitch skeletal muscle fibres. PMID- 20008469 TI - Can an NSAID a day keep muscle wasting away? PMID- 20008470 TI - Properties of the narrative scoring scheme using narrative retells in young school-age children. AB - PURPOSE: To evaluate the clinical utility of the narrative scoring scheme (NSS) as an index of narrative macrostructure for young school-age children. METHOD: Oral retells of a wordless picture book were elicited from 129 typically developing children, ages 5-7. A series of correlations and hierarchical regression equations were completed using microstructural measures of vocabulary and grammar to predict NSS scores. RESULTS: While the NSS was significantly correlated with age and each of the microstructural measures, the hierarchical regression analyses revealed a unique relationship between vocabulary and narrative macrostructure. CONCLUSION: The NSS is an efficient and informative tool for documenting children's development of narrative macrostructure. The relationship between the NSS and microstructural measures demonstrates that it is a robust measure of children's overall oral narrative competence and a powerful tool for clinicians and researchers. The unique relationship between lexical diversity and the NSS confirmed that a special relationship exists between vocabulary and narrative organization skills in young school-age children. PMID- 20008471 TI - Experienced speech-language pathologists' responses to ethical dilemmas: an integrated approach to ethical reasoning. AB - PURPOSE: To investigate the approaches of experienced speech-language pathologists (SLPs) to ethical reasoning and the processes they use to resolve ethical dilemmas. METHOD: Ten experienced SLPs participated in in-depth interviews. A narrative approach was used to guide participants' descriptions of how they resolved ethical dilemmas. Individual narrative transcriptions were analyzed by using the participant's words to develop an ethical story that described and interpreted their responses to dilemmas. Key concepts from individual stories were then coded into group themes to reflect participants' reasoning processes. RESULTS: Five major themes reflected participants' approaches to ethical reasoning: (a) focusing on the well-being of the client, (b) fulfilling professional roles and responsibilities, (c) attending to professional relationships, (d) managing resources, and (e) integrating personal and professional values. SLPs demonstrated a range of ethical reasoning processes: applying bioethical principles, casuistry, and narrative reasoning when managing ethical dilemmas in the workplace. CONCLUSIONS: The results indicate that experienced SLPs adopted an integrated approach to ethical reasoning. They supported clients' rights to make health care choices. Bioethical principles, casuistry, and narrative reasoning provided useful frameworks for facilitating health professionals' application of codes of ethics to complex professional practice issues. PMID- 20008472 TI - Development and initial validation of the localized scleroderma skin damage index and physician global assessment of disease damage: a proof-of-concept study. AB - OBJECTIVE: To develop and assess the psychometric properties of the Localized Scleroderma (LS) Skin Damage Index (LoSDI) and Physician Global Assessment of disease Damage (PGA-D). METHODS: Damage was defined as irreversible/persistent changes (>6 months) due to previous active disease/complications of therapy. Eight rheumatologists assessed the importance of 17 variables in formulating the PGA-D/LoSDI. LS patients were evaluated by two rheumatologists using both tools to assess their psychometric properties. LoSDI was calculated by summing three scores for cutaneous features of damage [dermal atrophy (DAT), subcutaneous atrophy (SAT) and dyspigmentation (DP)] measured at 18 anatomic sites. Patient GA of disease severity (PtGA-S), Children's Dermatology Life Quality Index (CDLQI) and PGA-D were recorded at the time of each examination. RESULTS: Thirty LS patients (112 lesions) and nine patient-visit pairs (18 lesions) were included for inter- and intra-rater reliability study. LoSDI and its domains DAT, SAT, DP and PGA-D demonstrated excellent inter- and intra-rater reliability (reliability coefficients 0.86-0.99 and 0.74-0.96, respectively). LoSDI correlated moderately with PGA-D and poorly with PtGA-S and CDLQI. PGA-D correlated moderately with PtGA-S, but poorly with CDLQI. CONCLUSIONS: To complete the LS Cutaneous Assessment Tool (LoSCAT), we developed and evaluated the psychometric properties of the LoSDI and PGA-D in addition to the LS Skin Severity Index (LoSSI). These instruments will facilitate evaluation of LS patients for individual patient management and clinical trials. LoSDI and PGA-D demonstrated excellent reliability and high validity. LoSCAT provides an improved understanding of LS natural history. Further study in a larger group of patients is needed to confirm these preliminary findings. PMID- 20008473 TI - Gastrointestinal manifestation of systemic sclerosis--thickening of the upper gastrointestinal wall detected by endoscopic ultrasound is a valid sign. AB - OBJECTIVE: Gastrointestinal (GI) tract involvement has been observed in the majority of patients with SSc. This has been attributed to an accumulation of extracellular matrix within the GI walls. We visualized the walls of the oesophagus, stomach and duodenum with its layers and measured the thickness in SSc patients and control patients utilizing endoscopic ultrasound (EUS). METHODS: Twenty-five SSc patients and 25 controls were evaluated. In addition to analysis of clinical symptoms, endoscopy and EUS (20-MHz miniprobe) were performed. The thickness of the complete wall was measured, and the mucosa, submucosa and muscularis were evaluated separately. RESULTS: Clinical symptoms of SSc patients were dysphagia (14/25) and heartburn (19/25). Endoscopic findings were hiatal hernia (16/25), oesophagitis (6/25), amotility (19/25) and a dehiscent pylorus (15/25). In comparison with controls, SSc patients had significantly thicker oesophageal [SSc 1.619 (0.454) mm, control 1.392 (0.333) mm; P = 0.025], antral [SSc 1.876 (0.635) mm, control 1.599 (0.291) mm; P = 0.029] and duodenal [SSc 1.730 (0.522) mm, control 1.525 (0.222) mm; P = 0.039] walls. Predominantly, submucosa and muscularis were significantly thicker in SSc patients. The presence of dysphagia or amotility was significantly associated with the thickening of the GI walls. CONCLUSIONS: The EUS revealed a significant thickening of the walls of the upper GI tract in SSc patients. Predominantly, the submucosa and muscularis are enlarged. These results strengthen the hypothesis that increased matrix deposition is an important aspect in the pathogenesis of GI involvement in SSc. PMID- 20008474 TI - Calcium-dependent regulation of voltage-gated sodium channels in cardiac myocytes: just the beginning? PMID- 20008475 TI - Exon skipping with morpholino oligomers: new treatment option for cardiomyopathy in Duchenne muscular dystrophy? PMID- 20008477 TI - sORF finder: a program package to identify small open reading frames with high coding potential. AB - SUMMARY: sORF finder is a program package for identifying small open reading frames (sORFs) with high-coding potential. This application allows the identification of coding sORFs according to the nucleotide composition bias among coding sequences and the potential functional constraint at the amino acid level through evaluation of synonymous and non-synonymous substitution rates. AVAILABILITY: Online tools and source codes are freely available at http://evolver.psc.riken.jp/. SUPPLEMENTARY INFORMATION: Supplementary data are available at Bioinformatics online. PMID- 20008476 TI - Exogenous and endogenous ceramides elicit volume-sensitive chloride current in ventricular myocytes. AB - AIMS: Because ceramide accumulates in several forms of cardiovascular disease and ceramide-induced apoptosis may involve the volume-sensitive Cl(-) current, I(Cl,swell), we assessed whether ceramide activates I(Cl,swell). METHODS AND RESULTS: I(Cl,swell) was measured in rabbit ventricular myocytes by whole-cell patch clamp after isolating anion currents. Exogenous C(2)-ceramide (C(2)-Cer), a membrane-permeant short-chain ceramide, elicited an outwardly rectifying Cl(-) current in both physiological and symmetrical Cl(-) solutions that was fully inhibited by DCPIB, a specific I(Cl,swell) blocker. In contrast, the metabolically inactive C(2)-Cer analogue C(2)-dihydroceramide (C(2)-H(2)Cer) failed to activate Cl(-) current. Bacterial sphingomyelinase (SMase), which generates endogenous long-chain ceramides as was confirmed by tandem mass spectrometry, also elicited an outwardly rectifying Cl(-) current that was inhibited by DCPIB and tamoxifen, another I(Cl,swell) blocker. Bacterial SMase induced current was partially reversed by osmotic shrinkage and fully suppressed by ebselen, a scavenger of reactive oxygen species. Outward rectification with physiological and symmetrical Cl(-) gradients, block by DCPIB and tamoxifen, and volume sensitivity are characteristics that identify I(Cl,swell). Insensitivity to C(2)-H(2)Cer and block by ebselen suggest involvement of ceramide signalling rather than direct lipid-channel interaction. CONCLUSION: Exogenous and endogenous ceramide elicited I(Cl,swell) in ventricular myocytes. This may contribute to persistent activation of I(Cl,swell) and aspects of altered myocyte function in cardiovascular diseases associated with by ceramide accumulation. PMID- 20008479 TI - Frontiers of research on organelle differentiation. PMID- 20008478 TI - Estimating DNA coverage and abundance in metagenomes using a gamma approximation. AB - MOTIVATION: Shotgun sequencing generates large numbers of short DNA reads from either an isolated organism or, in the case of metagenomics projects, from the aggregate genome of a microbial community. These reads are then assembled based on overlapping sequences into larger, contiguous sequences (contigs). The feasibility of assembly and the coverage achieved (reads per nucleotide or distinct sequence of nucleotides) depend on several factors: the number of reads sequenced, the read length and the relative abundances of their source genomes in the microbial community. A low coverage suggests that most of the genomic DNA in the sample has not been sequenced, but it is often difficult to estimate either the extent of the uncaptured diversity or the amount of additional sequencing that would be most efficacious. In this work, we regard a metagenome as a population of DNA fragments (bins), each of which may be covered by one or more reads. We employ a gamma distribution to model this bin population due to its flexibility and ease of use. When a gamma approximation can be found that adequately fits the data, we may estimate the number of bins that were not sequenced and that could potentially be revealed by additional sequencing. We evaluated the performance of this model using simulated metagenomes and demonstrate its applicability on three recent metagenomic datasets. SUPPLEMENTARY INFORMATION: Supplementary data are available at Bioinformatics online. PMID- 20008480 TI - Frontiers of research on work and retirement. AB - The shifting boundary between work and retirement and the always-emergent features of retirement practice create a wide opportunity for scholarship and research. After an overview of the scope of retirement research, this article articulates 4 areas that deserve special attention in the present historical circumstance: studies of the form and timing of retirement exits, the labor market for older workers, the quality of pensions, and the experience of retired life. The field should be wary of prescribing regimes of behavior for late careers and retirement that many people are unsuited to fulfill. PMID- 20008481 TI - Successful aging in the United States: prevalence estimates from a national sample of older adults. AB - OBJECTIVES: To estimate the prevalence of successful aging in the United States, with the broad aim of contributing to the dialogue on Rowe and Kahn's concept of successful aging. METHODS: Using data from the Health and Retirement Study, the prevalence of successful aging was calculated for adults aged 65 years and older at four time points: 1998, 2000, 2002, and 2004. Successful aging was operationalized in accordance with Rowe and Kahn's definition, which encompasses disease and disability, cognitive and physical functioning, social connections, and productive activities. RESULTS: No greater than 11.9% of older adults were aging "successfully" in any year. The adjusted odds of successful aging were generally lower for those of advanced age, male gender, and lower socioeconomic status. Between 1998 and 2004, the odds of successful aging declined by 25%, after accounting for demographic changes in the older population. DISCUSSION: Few older adults meet the criteria put forth in Rowe and Kahn's definition of successful aging, suggesting the need for modification if the concept is to be used for broad public health purposes. Disparities in successful aging were evident for socially defined subgroups, highlighting the importance of structural factors in enabling successful aging. PMID- 20008482 TI - How the Medicare Part D drug benefit changed the distribution of out-of-pocket pharmacy spending among older beneficiaries. AB - OBJECTIVE: To examine the impact of the Medicare drug benefit (Part D) on the distribution of out-of-pocket pharmacy spending among older adults. METHODS: We used a pre-post-with-comparison-group design to compare four groups of beneficiaries continuously enrolled in a Medicare Advantage plan between 2004 and 2007: three intervention groups with no or limited (quarterly caps of $150 or $350) prior coverage that obtained Part D benefits in 2006 and a comparison group with stable drug coverage from 2004 to 2007. RESULTS: The comparison group's out of-pocket drug spending was stable throughout 2004-2007, whereas Part D reduced out-of-pocket spending 13.4% among those without prior coverage (95% confidence interval [CI] -17.1% to -9.1%) and 15.9% among those with $150 quarterly caps (95% CI -19.1% to -12.8%) relative to the comparison group. Individuals in the top decile of drug spending paid a greater share of their costs out-of-pocket than others in the top 5 deciles. DISCUSSION: Although Part D reduced out-of pocket expenditures for older adults, those with the highest drug spending still pay a substantial share of their drug costs out-of-pocket. Thus, the Part D benefit does not achieve a primary purpose of insurance-to offer the greatest financial protection to those at the highest risk. PMID- 20008483 TI - Ethnicity and changing functional health in middle and late life: a person centered approach. AB - OBJECTIVES: Following a person-centered approach, this research aims to depict distinct courses of disability and to ascertain how the probabilities of experiencing these trajectories vary across Black, Hispanic, and White middle aged and older Americans. METHODS: Data came from the 1995-2006 Health and Retirement Study, which involved a national sample of 18,486 Americans older than 50 years of age. Group-based semiparametric mixture models (Proc Traj) were used for data analysis. RESULTS: Five trajectories were identified: (a) excellent functional health (61%), (b) good functional health with small increasing disability (25%), (c) accelerated increase in disability (7%), (d) high but stable disability (4%), and (e) persistent severe impairment (3%). However, when time-varying covariates (e.g., martial status and health conditions) were controlled, only 3 trajectories emerged: (a) healthy functioning (53%), moderate functional decrement (40%), and (c) large functional decrement (8%). Black and Hispanic Americans had significantly higher probabilities than White Americans in experiencing poor functional health trajectories, with Blacks at greater risks than Hispanics. CONCLUSIONS: Parallel to the concepts of successful aging, usual aging, and pathological aging, there exist distinct courses of changing functional health over time. The mechanisms underlying changes in disability may vary between Black and Hispanic Americans. PMID- 20008484 TI - Number of children and upstream intergenerational financial transfers: evidence from Hong Kong. AB - OBJECTIVES: This study examined financial transfers from adult children to elderly parents in Hong Kong and tested three hypotheses about the motives for such transfers. We address previous research, suggesting that family financial support for retirees will decline in the coming decades as a consequence of the reduction in the fertility rate; we also examine whether financial transfers are a function of the number of adult children in the family. METHODS: We used multiple regression models based on data from a representative sample of parents aged 60 years and older to identify the correlates of the amount of transfers from adult children to their elderly parents. RESULTS: We found evidence for the hypothesis that upstream transfers to elderly parents are their way of withdrawing savings from a "support bank" in which they made contribution for their children's education earlier in life and that transfers are altruistic in nature, but our results provide only moderate support to the old age security hypothesis that perceives family as a source of capital. DISCUSSION: The number of children has a ceiling effect on transfers, which calls into question common assumptions about the extent to which the decline in fertility will pose a severe threat to the extent of familial support of older persons over the coming decades. PMID- 20008485 TI - Social networks and well-being: a comparison of older people in Mediterranean and non-Mediterranean countries. AB - OBJECTIVES: This study examined whether the social networks of older persons in Mediterranean and non-Mediterranean countries were appreciably different and whether they functioned in similar ways in relation to well-being outcomes. METHODS: The sample included family household respondents aged 60 years and older from the first wave of the Survey of Health, Ageing and Retirement in Europe in 5 Mediterranean (n = 3,583) and 7 non-Mediterranean (n = 5,471) countries. Region was regressed separately by gender on variables from 4 network domains: structure and interaction, exchange, engagement and relationship quality, and controlling for background and health characteristics. In addition, 2 well-being outcomes depressive symptoms and perceived income inadequacy-were regressed on the study variables, including regional social network interaction terms. RESULTS: The results revealed differences across the 2 regional settings in each of the realms of social network, above and beyond the differences that exist in background characteristics and health status. The findings also showed that the social network variables had different effects on the well-being outcomes in the respective settings. DISCUSSION: The findings underscore that the social network phenomenon is contextually bound. The social networks of older people should be seen within their unique regional milieu and in relation to the values and social norms that prevail in different sets of societies. PMID- 20008486 TI - Selections that optimize RNA display in the yeast three-hybrid system. AB - The yeast three-hybrid system (Y3H) is a powerful tool to select or confirm RNA protein interactions. Target protein recognition of an RNA insert within a test transcript depends on at least three factors: intrinsic protein affinity for the properly folded insert, retention of RNA insert tertiary structure within a longer RNA transcript, and accessibility of the RNA insert to the target protein. Y3H reporter gene readout reflects the combination of these factors. Here, we discuss RNA insert tertiary structure and accessibility in the Y3H as "RNA display." We review evidence that RNA display can sometimes be optimized during Y3H selections that do not increase the intrinsic affinity of an RNA insert for a target protein. This situation is more likely when a library of RNA inserts and heterogeneous flanking sequences is subjected to selection, and is less likely when point mutations are targeted to the insert in a fixed context. An RNA display vector with enhanced modularity has been developed to minimize sequence context effects in the Y3H. PMID- 20008487 TI - Phosphorylation regulates removal of synaptic N-methyl-D-aspartate receptors after withdrawal from chronic ethanol exposure. AB - Alterations in N-methyl-d-aspartate receptor (NMDAR) protein levels or subcellular localization in brain after chronic ethanol exposure may contribute to withdrawal-associated seizures and neurotoxicity. We have investigated synaptic localization of NMDARs in cultured hippocampal pyramidal neurons after prolonged (7 days) exposure to, and acute withdrawal from, 80 mM ethanol using fluorescence immunocytochemistry techniques. After chronic ethanol exposure, there was a significant increase in the clustering of NR1 and NR2B subunits and their colocalization with the synaptic proteins synaptophysin and postsynaptic density protein 95, respectively. There was also increased expression of NR1 variants containing the C2' cassette after chronic ethanol exposure. The ethanol induced synaptic clustering and colocalization were rapidly reversed within 4 h after ethanol withdrawal. Surface labeling of NR2B subunits suggested that this rapid reversal involved lateral receptor movement to extrasynaptic sites rather than internalization of receptors. Receptor removal from the synapse during ethanol withdrawal was associated with changes in the phosphorylation state of NR2B Ser1480, controlled by the protein kinase CK2. The redistribution of NMDAR to synapses produced by long-term ethanol exposure, as well as the rapid removal during withdrawal, may not only affect neuronal withdrawal hyperexcitability but also may sensitize the system to subsequent synaptic plasticity. PMID- 20008488 TI - Up-regulation of histamine H4 receptors contributes to splenic apoptosis in septic mice: counteraction of the antiapoptotic action of nuclear factor-kappaB. AB - The histamine H(4) receptor is the most recently identified receptor and is considered to play a role in a variety of inflammatory diseases. Histamine levels in the plasma are known to be elevated in animal models of sepsis and in septic patients. The aim of this study was to test the hypothesis that the H(4) receptor may play a significant role in the pathophysiology of sepsis. Polymicrobial sepsis was induced by cecal ligation and puncture in BALB/c mice. Although the H(4) receptor gene was undetectable in normal peripheral key organs, with the exception of the spleen, the expression levels of this gene were highly up regulated in all those organs of septic mice. In vivo transfection of nuclear factor-kappaB (NF-kappaB) decoy oligodeoxynucleotide, but not of its scrambled form, resulted in a great inhibition of sepsis-induced overexpression of the H(4) receptor gene. In septic mice, marked increases in caspase-3 activation and follicular lymphocyte apoptosis in spleens were strongly suppressed by systemic treatment with synthetic small interfering RNA (siRNA) targeted to the H(4) receptor. This was associated with the up-regulation of a number of antiapoptotic proteins. These antiapoptotic effects of H(4) receptor siRNA treatment were all inhibited by further application of NF-kappaB decoy oligonucleotide. Our results suggest that superinduction of the histamine H(4) receptor gene in peripheral key organs, including the spleen, that is promoted by sepsis is transcriptionally controlled by NF-kappaB, whereas stimulation of this receptor is involved in the development of sepsis-induced splenic apoptosis through counteraction of the antiapoptotic action of NF-kappaB. PMID- 20008489 TI - The effect of protein kinase C and G protein-coupled receptor kinase inhibition on tolerance induced by mu-opioid agonists of different efficacy. AB - Differences in the mechanisms underlying tolerance and mu-opioid receptor desensitization resulting from exposure to opioid agonists of different efficacy have been suggested previously. The objective of this study was to determine the effects of protein kinase C (PKC) and G protein-coupled receptor kinase (GRK) inhibition on antinociceptive tolerance in vivo to opioid agonists of different efficacy. A rapid (8-h) tolerance-induction model was used where each opioid was repeatedly administered to naive mice. Animals were then challenged with the opioid after injection of a kinase inhibitor to determine its effects on the level of tolerance. Tolerance to meperidine, morphine, or fentanyl was fully reversed by the PKC inhibitor 12-(2-cyanoethyl)-6,7,12,13-tetrahydro-13-methyl-5 oxo-5H-indolo(2,3-a)pyrrolo(3,4-c)carbazole (Go6976). However, in vivo tolerance to [d-Ala(2),N-Me-Phe(4),Gly(5)-ol]-enkephalin (DAMGO) was not reversed by PKC inhibition. The novel small-molecule GRK inhibitors beta-adrenergic receptor kinase 1 inhibitor and 2-(8-[(dimethylamino) methyl]-6,7,8,9 tetrahydropyridol[1,2-a]indol-3-yl)-3-(1-methylindol-3-yl)maleimide (Ro 32-0432) did not reverse the tolerance to meperidine, fentanyl, or morphine but did reverse the tolerance to DAMGO. To correlate GRK-dependent DAMGO-induced tolerance with mu-opioid receptor desensitization, we used in vitro whole-cell patch-clamp recording from mouse locus coeruleus neurons and observed that the GRK inhibitors reduced DAMGO-induced desensitization of mu-opioid receptors, whereas the PKC inhibitor had no effect. These results suggest that tolerance induced by low- and moderate-efficacy mu-opioid receptor agonists is dependent on PKC, whereas tolerance induced by the high-efficacy agonist DAMGO is dependent on GRK. PMID- 20008490 TI - Atrial fibrillation ablation in systolic heart failure patients: a promising tool? PMID- 20008491 TI - Atrial fibrillation ablation in athletes: an Olympic challenge? PMID- 20008492 TI - Positive cultures in asymptomatic patients during elective device replacement: a murderer hides in the darkness or an innocent person on the crime scene? PMID- 20008493 TI - Atrial arrhythmias after cardiac resynchronization therapy: an inverse correlation with achieving 100% biventricular pacing and cardiac resynchronization therapy effectiveness. PMID- 20008494 TI - Ventricular pacing: to pace or not to pace. PMID- 20008495 TI - Device implantation: the human factor. PMID- 20008497 TI - Detection of T-wave alternans by exercise stress testing, atrial-pacing, and atrioventricular pacing. PMID- 20008498 TI - CIDA adopts new food security strategy. PMID- 20008499 TI - Physicians' legal duty of care and legal right to refuse to work during a pandemic. PMID- 20008501 TI - Public health: Exposure to bats: updated recommendations. PMID- 20008502 TI - Mees lines and Beau lines after chemotherapy. PMID- 20008504 TI - Liberal senators assail consumer safety bill. PMID- 20008503 TI - Serum anion gap, bicarbonate and biomarkers of inflammation in healthy individuals in a national survey. AB - BACKGROUND: In vitro data suggest that lower extracellular pH activates the immune system. We conducted a population-based study of the relation between serum acid-base status and inflammation. METHODS: We examined the serum anion gap and serum levels of bicarbonate and inflammatory biomarkers in 4525 healthy adults who participated in the National Health and Nutrition Examination Survey during 1999-2006. We excluded participants who had chronic disease, recent infection and an estimated glomerular filtration rate of less than 60 mL/min per 1.73 m2. RESULTS: The mean values of serum anion gap, bicarbonate level, leukocyte count and C-reactive protein level were all within normal limits. After adjustment for age, sex, ethnic background, body mass index, serum albumin level and other factors, we found that a higher anion gap and lower bicarbonate level were associated with a higher leukocyte count and higher C-reactive protein level. Compared with participants in the lowest quartile of anion gap, those in the highest quartile had a leukocyte count that was 1.0x10(9)/L higher and a C reactive protein level that was 10.9 nmol/L higher (p<0.01). Compared with participants in the highest quartile of bicarbonate level, those in the lowest quartile had a leukocyte count that was 0.7x10(9)/L higher and a C-reactive protein level that was 4.0 nmol/L higher (p 0.8 poly(A)*poly(U) does not disproportionate at 25 degrees C and dye intercalation into AU to give AUD is the only observed process. Melting experiments as well show that coralyne induces the duplex disproportionation. Effects of temperature, ionic strength and ethanol content are investigated. One concludes that triplex formation requires coralyne be only partially intercalated into AUD. Under suitable concentration conditions, this feature favours the interaction of free AU with AUD to give the AUDAU intermediate which evolves into triplex UAUD and single-stranded poly(A). Duplex poly(A)*poly(A) undergoes aggregation as well, but only at much higher polymer concentrations compared to poly(A)*poly(U). PMID- 20008510 TI - Imprecise transcription termination within Escherichia coli greA leader gives rise to an array of short transcripts, GraL. AB - We report that greA expression is driven by two strong, overlapping P1 and P2 promoters. The P1 promoter is sigma(70)-dependent and P2 is sigma(E)-dependent. Two-thirds of transcripts terminate within the leader region and the remaining third comprises greA mRNA. Termination efficiency seems to be unaffected by growth phase. Two collections of small 40-50 (initiating from P2) and 50-60 nt (from P1) RNA chains, termed GraL, are demonstrable in vivo and in vitro. We document that GraL arrays arise from an intrinsic terminator with an 11 bp stem followed by an AU(7)GCU(2) sequence. Atypical chain termination occurs at multiple sites; the 3'-ends differ by 1 nt over a range of 10 nt. Transcripts observed are shown to be insensitive to Gre factors and physically released from RNAP-DNA complexes. The abundance of individual chains within each cluster displays a characteristic pattern, which can be differentially altered by oligonucleotide probes. Multiple termination sites are particularly sensitive to changes at the bottom of the stem. Evolutionarily conserved GraL stem structures and fitness assays suggest a biological function for the RNA clusters themselves. Although GraL overexpression induces >/=3-fold transcriptional changes of over 100 genes, a direct target remains elusive. PMID- 20008511 TI - Histone chaperone Chz1p regulates H2B ubiquitination and subtelomeric anti silencing. AB - Chz1p is a histone chaperone that interacts physically and functionally with the histone variant Htz1p, which has been implicated in establishing and maintaining boundaries between transcriptionally inactive heterochromatin and active euchromatin. To investigate the role of Chz1p in chromatin organization, we performed genome-wide expression arrays and chromatin immunoprecipitations of SIR complex components and modified histones in a CHZ1 deletion strain. Deletion of CHZ1 led to reduced ubiquitination of subtelomere-associated H2B, reduced subtelomeric H3K79 di-methylation, and increased binding of Sir3p, and Sir4p at telomere-distal euchromatin regions, correlating with decreased gene expression in subtelomeric regions. This anti-silencing defect appears to be mediated by enhanced association of de-ubiquitinase Ubp10p with subtelomeric DNA, as detected by chromatin immunoprecipitation analysis. In support of this, we show that deletion of UBP10 can antagonize the subtelomeric silencing phenotype of Deltachz1. Taken together, the results demonstrate a novel role for Chz1p in epigenetic regulation, through H2B de-ubiquitination by Ubp10p. PMID- 20008513 TI - SoyBase, the USDA-ARS soybean genetics and genomics database. AB - SoyBase, the USDA-ARS soybean genetic database, is a comprehensive repository for professionally curated genetics, genomics and related data resources for soybean. SoyBase contains the most current genetic, physical and genomic sequence maps integrated with qualitative and quantitative traits. The quantitative trait loci (QTL) represent more than 18 years of QTL mapping of more than 90 unique traits. SoyBase also contains the well-annotated 'Williams 82' genomic sequence and associated data mining tools. The genetic and sequence views of the soybean chromosomes and the extensive data on traits and phenotypes are extensively interlinked. This allows entry to the database using almost any kind of available information, such as genetic map symbols, soybean gene names or phenotypic traits. SoyBase is the repository for controlled vocabularies for soybean growth, development and trait terms, which are also linked to the more general plant ontologies. SoyBase can be accessed at http://soybase.org. PMID- 20008514 TI - Defining hyperandrogenism in polycystic ovary syndrome: measurement of testosterone and androstenedione by liquid chromatography-tandem mass spectrometry and analysis by receiver operator characteristic plots. AB - OBJECTIVE: Hyperandrogenism is one of the diagnostic criteria for the polycystic ovary syndrome (PCOS) despite no agreed definition of hyperandrogenism. In part, this is due to the quality of testosterone immunoassays. We have developed liquid chromatography-tandem mass spectrometry methods for analysing testosterone and androstenedione (Ad) to study their reference ranges and diagnostic utility in PCOS. DESIGN, SETTING AND SUBJECTS: A consecutive series of 122 women attending a reproductive medicine clinic. METHODS: Blood samples were taken during the early follicular phase for measurement of LH, FSH, oestradiol, Ad, testosterone and sex hormone-binding globulin (SHBG). Retrospective case note analysis was used to determine the clinical features and ultrasound findings. RESULTS: The incidence of PCOS was 13.9%. The reference interval for testosterone was <1.8 nmol/l and for Ad was 1.4-7.4 nmol/l. There were significant differences in total testosterone (P=0.001), Ad (P<0.05) and free androgen index (FAI; P<0.0001) between the women with and without PCOS. Diagnostic performance using receiver operator characteristic plots showed area under the curve (AUC) for FAI 0.81, testosterone 0.75 and Ad 0.66. The AUC for the LH:FSH ratio was 0.72. CONCLUSIONS: Our analysis of a consecutive series of women attending a reproductive clinic has provided an appropriate series on which to construct reference ranges for key androgens in women. Secondly, it has allowed us to conclude that early follicular serum testosterone measured using tandem mass spectrometry, FAI and the LH:FSH ratio are valuable laboratory tests in the diagnosis of PCOS. PMID- 20008512 TI - CK2 phosphorylation-dependent interaction between aprataxin and MDC1 in the DNA damage response. AB - Aprataxin, defective in the neurodegenerative disorder ataxia oculomotor apraxia type 1, resolves abortive DNA ligation intermediates during DNA repair. Here, we demonstrate that aprataxin localizes at sites of DNA damage induced by high LET radiation and binds to mediator of DNA-damage checkpoint protein 1 (MDC1/NFBD1) through a phosphorylation-dependent interaction. This interaction is mediated via the aprataxin FHA domain and multiple casein kinase 2 di-phosphorylated S-D-T-D motifs in MDC1. X-ray structural and mutagenic analysis of aprataxin FHA domain, combined with modelling of the pSDpTD peptide interaction suggest an unusual FHA binding mechanism mediated by a cluster of basic residues at and around the canonical pT-docking site. Mutation of aprataxin FHA Arg29 prevented its interaction with MDC1 and recruitment to sites of DNA damage. These results indicate that aprataxin is involved not only in single strand break repair but also in the processing of a subset of double strand breaks presumably through its interaction with MDC1. PMID- 20008515 TI - Pleiotropic effects of YC-1 selectively inhibit pathological retinal neovascularization and promote physiological revascularization in a mouse model of oxygen-induced retinopathy. AB - Vascular endothelial growth factor (VEGF) and inducible nitric-oxide synthase (iNOS) have been implicated in ischemia-induced retinal neovascularization. Retinal ischemia has been shown to induce VEGF and iNOS expression. It has been postulated that one of the crucial consequences of iNOS expression in the ischemic retina is the inhibition of angiogenesis. Furthermore, iNOS was shown to be overexpressed in Muller cells from patients with diabetic retinopathy. YC-1, a small molecule inhibitor of hypoxia-inducible factor (HIF)-1 alpha, has been shown to inhibit iNOS expression in various tissue models. Our aim was to assess the pleiotropic effects of YC-1 in an oxygen-induced retinopathy (OIR) mouse model and evaluate its therapeutic potential in HIF-1- and iNOS-mediated retinal pathologies. Dual-injections of YC-1 into the neovascular retinas decreased the total retinopathy score, inhibited vaso-obliteration and pathologic tuft formation, and concomitantly promoted physiological retinal revascularization, compared with dimethyl sulfoxide (DMSO)-treated group. Furthermore, YC-1-treated retinas exhibited a marked increase in immunoreactivities for CD31 and von Willebrand factor and displayed significant inhibition in HIF-1alpha protein expression. Furthermore, YC-1 down-regulated VEGF, erythropoietin, endothelin-1, matrix metalloproteinase-9, and iNOS message and protein levels. When hypoxic Muller and neuoroglial cells were treated with YC-1, iNOS mRNA and protein levels were reduced in a dose-dependent fashion. We demonstrate that YC-1 inhibits pathological retinal neovascularization by exhibiting antineovascular activities, which impaired ischemia-induced expression of HIF-1 and its downstream angiogenic molecules. Furthermore, YC-1 enhanced physiological revascularization of the retinal vascular plexuses via the inhibition of iNOS mRNA and protein expressions. The pleiotropic effects of YC-1 allude to its possible use as a promising therapeutic iNOS inhibitor candidate for the treatment of retinal neovascularization. PMID- 20008517 TI - beta-D-Galactosidase from Paenibacillus thiaminolyticus catalyzing transfucosylation reactions. AB - A genomic library of bacterial strain Paenibacillus thiaminolyticus was constructed and the plasmid DNA of the clone, containing the gene encoding beta-d galactosidase with beta-d-fucosidase activity, detected by 5-bromo-4-chloro-3 indoxyl beta-d-galactopyranoside, was sequenced. Cells of Escherichia coli BL21 (DE3) were used for production of the enzyme in the form of a histidine-tagged protein. This recombinant fusion protein was purified using Ni-NTA agarose affinity chromatography and characterized by using p-nitrophenyl beta-d fucopyranoside (K(m) value of (1.18 +/- 0.06) mmol/L), p-nitrophenyl beta-d galactopyranoside (K(m) value of (250 +/- 40) mmol/L), p-nitrophenyl beta-d glucopyranoside (K(m) value of (77 +/- 6) mmol/L), and lactose (K(m) value of (206 +/- 5) mmol/L) as substrates. Optimal pH and temperature were estimated as 5.5 and 65 degrees C, respectively. According to the amino acid sequence, the molecular weight of the fusion protein was calculated to be 68.6 kDa and gel filtration chromatography confirmed the presence of the enzyme in a monomeric form. In the following step, its ability to catalyze transfucosylation reactions was tested. The enzyme was able to catalyze the transfer of fucosyl moiety to different p-nitrophenyl glycopyranosides (producing p-nitrophenyl beta-d fucopyranosyl-(1,3)-beta-d-fucopyranoside, p-nitrophenyl beta-d-fucopyranosyl (1,3)-alpha-d-glucopyranoside, p-nitrophenyl beta-d-fucopyranosyl-(1,3)-alpha-d mannopyranoside, and p-nitrophenyl beta-d-fucopyranosyl-(1,6)-alpha-d galactopyranoside) and alcohols (producing methyl beta-d-fucopyranoside, ethyl beta-d-fucopyranoside, 1-propyl beta-d-fucopyranoside, 2-propyl beta-d fucopyranoside, 1-octyl beta-d-fucopyranoside, and 2-octyl beta-d fucopyranoside). These results indicate the possibility of utilizing this enzyme as a promising tool for enzymatic synthesis of beta-d-fucosylated molecules. PMID- 20008516 TI - Simple 2,4-diacylphloroglucinols as classic transient receptor potential-6 activators--identification of a novel pharmacophore. AB - The naturally occurring acylated phloroglucinol derivative hyperforin was recently identified as the first specific canonical transient receptor potential 6 (TRPC6) activator. Hyperforin is the major antidepressant component of St. John's wort, which mediates its antidepressant-like properties via TRPC6 channel activation. However, its pharmacophore moiety for activating TRPC6 channels is unknown. We hypothesized that the phloroglucinol moiety could be the essential pharmacophore of hyperforin and that its activity profile could be due to structural similarities with diacylglycerol (DAG), an endogenous nonselective activator of TRPC3, TRPC6, and TRPC7. Accordingly, a few 2-acyl and 2,4 diacylphloroglucinols were tested for their hyperforin-like activity profiles. We used a battery of experimental models to investigate all functional aspects of TRPC6 activation, including ion channel recordings, Ca(2+) imaging, neurite outgrowth, and inhibition of synaptosomal uptake. Phloroglucinol itself was inactive in all of our assays, which was also the case for 2-acylphloroglucinols. For TRPC6 activation, the presence of two symmetrically acyl-substitutions with appropriate alkyl chains in the phloroglucinol moiety seems to be an essential prerequisite. Potencies of these compounds in all assays were comparable with that of hyperforin for activating the TRPC6 channel. Finally, using structure based modeling techniques, we suggest a binding mode for hyperforin to TRPC6. Based on this modeling approach, we propose that DAG is able to activate TRPC3, TRPC6, and TRPC7 because of higher flexibility within the chemical structure of DAG compared with the rather rigid structures of hyperforin and the 2,4 diacylphloroglucinol derivatives. PMID- 20008518 TI - Flux regulation of cardiac ryanodine receptor channels. AB - The cardiac type 2 ryanodine receptor (RYR2) is activated by Ca2+-induced Ca2+ release (CICR). The inherent positive feedback of CICR is well controlled in cells, but the nature of this control is debated. Here, we explore how the Ca2+ flux (lumen-to-cytosol) carried by an open RYR2 channel influences its own cytosolic Ca2+ regulatory sites as well as those on a neighboring channel. Both flux-dependent activation and inhibition of single channels were detected when there were super-physiological Ca2+ fluxes (>3 pA). Single-channel results indicate a pore inhibition site distance of 1.2 +/- 0.16 nm and that the activation site on an open channel is shielded/protected from its own flux. Our results indicate that the Ca2+ flux mediated by an open RYR2 channel in cells (approximately 0.5 pA) is too small to substantially regulate (activate or inhibit) the channel carrying it, even though it is sufficient to activate a neighboring RYR2 channel. PMID- 20008521 TI - Host-dependent Lewis (Le) antigen expression in Helicobacter pylori cells recovered from Leb-transgenic mice. AB - Variation of surface antigen expression is a mechanism used by microbes to adapt to and persist within their host habitats. Helicobacter pylori, a persistent bacterial colonizer of the human stomach, can alter its surface Lewis (Le) antigen expression. We examined H. pylori colonization in mice to test the hypothesis that host phenotype selects for H. pylori (Le) phenotypes. When wild type and Le(b)-expressing transgenic FVB/N mice were challenged with H. pylori strain HP1, expressing Le(x) and Le(y), we found that bacterial populations recovered after 8 mo from Le(b)-transgenic, but not wild-type, mice expressed Le(b). Changes in Le phenotype were linked to variation of a putative galactosyltransferase gene (beta-(1,3)galT); mutagenesis and complementation revealed its essential role in type I antigen expression. These studies indicate that H. pylori evolves to resemble the host's gastric Le phenotype, and reveal a bacterial genetic locus that is subject to host-driven selection pressure. PMID- 20008522 TI - PD-L1 regulates the development, maintenance, and function of induced regulatory T cells. AB - Both the programmed death (PD) 1-PD-ligand (PD-L) pathway and regulatory T (T reg) cells are instrumental to the maintenance of peripheral tolerance. We demonstrate that PD-L1 has a pivotal role in regulating induced T reg (iT reg) cell development and sustaining iT reg cell function. PD-L1(-/-) antigen presenting cells minimally convert naive CD4 T cells to iT reg cells, showing the essential role of PD-L1 for iT reg cell induction. PD-L1-coated beads induce iT reg cells in vitro, indicating that PD-L1 itself regulates iT reg cell development. Furthermore, PD-L1 enhances and sustains Foxp3 expression and the suppressive function of iT reg cells. The obligatory role for PD-L1 in controlling iT reg cell development and function in vivo is illustrated by a marked reduction in iT reg cell conversion and rapid onset of a fatal inflammatory phenotype in PD-L1(-/-)PD-L2(-/-) Rag(-/-) recipients of naive CD4 T cells. PD-L1 iT reg cell development is mediated through the down-regulation of phospho-Akt, mTOR, S6, and ERK2 and concomitant with the up-regulation of PTEN, all key signaling molecules which are critical for iT reg cell development. Thus, PD-L1 can inhibit T cell responses by promoting both the induction and maintenance of iT reg cells. These studies define a novel mechanism for iT reg cell development and function, as well as a new strategy for controlling T reg cell plasticity. PMID- 20008523 TI - HLA-B*35-Px-mediated acceleration of HIV-1 infection by increased inhibitory immunoregulatory impulses. AB - A subset of HLA-B*35 alleles, B*35-Px, are strongly associated with accelerated HIV-1 disease progression for reasons that are not understood. Interestingly, the alternative set of B*35 subtypes, B*35-PY, have no detectable impact on HIV-1 disease outcomes, even though they can present identical HIV-1 epitopes as B*35 Px molecules. Thus, the differential impact of these alleles on HIV-1 disease progression may be unrelated to interactions with HIV-1-specific CD8(+) T cells. Here, we show that the B*35-Px molecule B*3503 binds with greater affinity to immunoglobulin-like transcript 4 (ILT4), an inhibitory MHC class I receptor expressed on dendritic cells, than does the B*35-PY molecule B*3501, even though these two B*35 molecules differ by only one amino acid and present identical HIV 1 epitopes. The preferential recognition of B*3503 by ILT4 was associated with significantly stronger dendritic cell dysfunction in in vitro functional assays. Moreover, HIV-1-infected carriers of B*3503 had poor dendritic cell functional properties in ex vivo assessments when compared with carriers of the B*3501 allele. Differential interactions between HLA class I allele subtypes and immunoregulatory MHC class I receptors on dendritic cells thus provide a novel perspective for the understanding of MHC class I associations with HIV-1 disease progression and for the manipulation of host immunity against HIV-1. PMID- 20008524 TI - Intestinal CD103+, but not CX3CR1+, antigen sampling cells migrate in lymph and serve classical dendritic cell functions. AB - Chemokine receptor CX3CR1(+) dendritic cells (DCs) have been suggested to sample intestinal antigens by extending transepithelial dendrites into the gut lumen. Other studies identified CD103(+) DCs in the mucosa, which, through their ability to synthesize retinoic acid (RA), appear to be capable of generating typical signatures of intestinal adaptive immune responses. We report that CD103 and CX3CR1 phenotypically and functionally characterize distinct subsets of lamina propria cells. In contrast to CD103(+) DC, CX3CR1(+) cells represent a nonmigratory gut-resident population with slow turnover rates and poor responses to FLT-3L and granulocyte/macrophage colony-stimulating factor. Direct visualization of cells in lymph vessels and flow cytometry of mouse intestinal lymph revealed that CD103(+) DCs, but not CX3CR1-expressing cells, migrate into the gut draining mesenteric lymph nodes (LNs) under steady-state and inflammatory conditions. Moreover, CX3CR1(+) cells displayed poor T cell stimulatory capacity in vitro and in vivo after direct injection of cells into intestinal lymphatics and appeared to be less efficient at generating RA compared with CD103(+) DC. These findings indicate that selectively CD103(+) DCs serve classical DC functions and initiate adaptive immune responses in local LNs, whereas CX3CR1(+) populations might modulate immune responses directly in the mucosa and serve as first line barrier against invading enteropathogens. PMID- 20008525 TI - Mincle is a long sought receptor for mycobacterial cord factor. AB - Mycobacterium tuberculosis is a leading killer worldwide, yet the adjuvancy of its cell wall has proven to be a valuable therapeutic tool for vaccination and immunotherapy. Much research effort has focused on the mycobacterial glycolipid trehalose-6,6'-dimycolate (TDM), a potent immunostimulant that is also known as cord factor. Now, the identification of the monocyte-inducible C-type lectin (Mincle) as an essential receptor for TDM provides new insight into the formation of the characteristic granulomas in tuberculosis and an avenue for rational adjuvant design. PMID- 20008526 TI - Direct recognition of the mycobacterial glycolipid, trehalose dimycolate, by C type lectin Mincle. AB - Tuberculosis remains a fatal disease caused by Mycobacterium tuberculosis, which contains various unique components that affect the host immune system. Trehalose 6,6'-dimycolate (TDM; also called cord factor) is a mycobacterial cell wall glycolipid that is the most studied immunostimulatory component of M. tuberculosis. Despite five decades of research on TDM, its host receptor has not been clearly identified. Here, we demonstrate that macrophage inducible C-type lectin (Mincle) is an essential receptor for TDM. Heat-killed mycobacteria activated Mincle-expressing cells, but the activity was lost upon delipidation of the bacteria; analysis of the lipid extracts identified TDM as a Mincle ligand. TDM activated macrophages to produce inflammatory cytokines and nitric oxide, which are completely suppressed in Mincle-deficient macrophages. In vivo TDM administration induced a robust elevation of inflammatory cytokines in sera and characteristic lung inflammation, such as granuloma formation. However, no TDM induced lung granuloma was formed in Mincle-deficient mice. Whole mycobacteria were able to activate macrophages even in MyD88-deficient background, but the activation was significantly diminished in Mincle/MyD88 double-deficient macrophages. These results demonstrate that Mincle is an essential receptor for the mycobacterial glycolipid, TDM. PMID- 20008527 TI - Bordetella evades the host immune system by inducing IL-10 through a type III effector, BopN. AB - The inflammatory response is one of several host alert mechanisms that recruit neutrophils from the circulation to the area of infection. We demonstrate that Bordetella, a bacterial pathogen, exploits an antiinflammatory cytokine, interleukin-10 (IL-10), to evade the host immune system. We identified a Bordetella effector, BopN, that is translocated into the host cell via the type III secretion system, where it induces enhanced production of IL-10. Interestingly, the BopN effector translocates itself into the nucleus and is involved in the down-regulation of mitogen-activated protein kinases. Using pharmacological blockade, we demonstrated that BopN-induced IL-10 production is mediated, at least in part, by its ability to block the extracellular signal regulated kinase pathway. We also showed that BopN blocks nuclear translocation of nuclear factor kappaB p65 (NF-kappaBp65) but, in contrast, promotes nuclear translocation of NF-kappaBp50. A BopN-deficient strain was unable to induce IL-10 production in mice, resulting in the elimination of bacteria via neutrophil infiltration into the pulmonary alveoli. Furthermore, IL-10-deficient mice effectively eliminated wild-type as well as BopN mutant bacteria. Thus, Bordetella exploits BopN as a stealth strategy to shut off the host inflammatory reaction. These results explain the ability of Bordetella species to avoid induction of the inflammatory response. PMID- 20008528 TI - The origin and development of nonlymphoid tissue CD103+ DCs. AB - CD103(+) dendritic cells (DCs) in nonlymphoid tissues are specialized in the cross-presentation of cell-associated antigens. However, little is known about the mechanisms that regulate the development of these cells. We show that two populations of CD11c(+)MHCII(+) cells separated on the basis of CD103 and CD11b expression coexist in most nonlymphoid tissues with the exception of the lamina propria. CD103(+) DCs are related to lymphoid organ CD8(+) DCs in that they are derived exclusively from pre-DCs under the control of fms-like tyrosine kinase 3 (Flt3) ligand, inhibitor of DNA protein 2 (Id2), and IFN regulatory protein 8 (IRF8). In contrast, lamina propria CD103(+) DCs express CD11b and develop independently of Id2 and IRF8. The other population of CD11c(+)MHCII(+) cells in tissues, which is CD103(-)CD11b(+), is heterogenous and depends on both Flt3 and MCSF-R. Our results reveal that nonlymphoid tissue CD103(+) DCs and lymphoid organ CD8(+) DCs derive from the same precursor and follow a related differentiation program. PMID- 20008529 TI - Fusobacterium nucleatum envelope protein FomA is immunogenic and binds to the salivary statherin-derived peptide. AB - We have previously shown that one of the minimal active regions of statherin, a human salivary protein, for binding to Fusobacterium nucleatum is a YQPVPE amino acid sequence. In this study, we identified the FomA protein of F. nucleatum, which is responsible for binding to the statherin-derived YQPVPE peptide. Overlay analysis showed that a 40-kDa protein of the F. nucleatum cell envelope (40-kDa CE) specifically bound to the YQPVPE peptide. The equilibrium association constant between the affinity-purified 40-kDa CE and the YQPVPE peptide was 4.30 x 10(6). Further, the purity and amino acid sequence analyses of the purified 40 kDa CE revealed approximately 98.7% (wt/wt) purity and a high degree of homology with FomA, a major porin protein of F. nucleatum. Thus, a FomA-deficient mutant failed to bind to the YQPVPE peptide. In addition, increased levels of a FomA specific mucosal IgA antibody (Ab) and plasma IgG and IgA Abs were seen only in mice immunized nasally with cholera toxin (CT) and the purified 40-kDa FomA protein. Interestingly, saliva from mice that received FomA plus CT as a mucosal adjuvant nasally prevented in vitro binding of F. nucleatum to statherin-coated polyvinyl chloride plates. Taken together, these results suggest that induction of specific immunity to the 40-kDa FomA protein of F. nucleatum, which specifically binds to the statherin-derived peptide, may be an effective tool for preventing the formation of F. nucleatum biofilms in the oral cavity. PMID- 20008530 TI - Pseudomonas aeruginosa-mediated damage requires distinct receptors at the apical and basolateral surfaces of the polarized epithelium. AB - Pseudomonas aeruginosa, an important opportunistic pathogen of humans, exploits epithelial damage to establish infection. We have rigorously explored the role of N-glycoproteins and heparan sulfate proteoglycans (HSPGs) in P. aeruginosa mediated attachment and subsequent downstream events at the apical (AP) and basolateral (BL) surfaces of polarized epithelium. We demonstrate that the N glycan chains at the AP surface are necessary and sufficient for binding, invasion, and cytotoxicity to kidney (MDCK) and airway (Calu-3) cells grown at various states of polarization on Transwell filters. Upregulation of N glycosylation enhanced binding, whereas pharmacologic inhibition of N glycosylation or infection of MDCK cells defective in N-glycosylation resulted in decreased binding. In contrast, at the BL surface, the HS moiety of HSPGs mediated P. aeruginosa binding, cytotoxicity, and invasion. In incompletely polarized epithelium, HSPG abundance was increased at the AP surface, explaining its increased susceptibility to P. aeruginosa colonization and damage. Using MDCK cells grown as three-dimensional cysts as a model for epithelial organs, we show that P. aeruginosa specifically colocalized with HS-rich areas at the BL membrane but with complex N-glycans at the AP surface. Finally, P. aeruginosa bound to HS chains and N-glycans coated on plastic surfaces, showing the highest binding affinity toward isolated HS chains. Together, these findings demonstrate that P. aeruginosa recognizes distinct receptors on the AP and BL surfaces of polarized epithelium. Changes in the composition of N-glycan chains and/or in the distribution of HSPGs may explain the enhanced susceptibility of damaged epithelium to P. aeruginosa. PMID- 20008531 TI - Regulatory role of the MisR/S two-component system in hemoglobin utilization in Neisseria meningitidis. AB - Outer membrane iron receptors are some of the major surface entities that are critical for meningococcal pathogenesis. The gene encoding the meningococcal hemoglobin receptor, HmbR, is both independently transcribed and transcriptionally linked to the upstream gene hemO, which encodes a heme oxygenase. The MisR/S two-component system was previously determined to regulate hmbR transcription, and its hemO and hmbR regulatory mechanisms were characterized further here. The expression of hemO and hmbR was downregulated in misR/S mutants under both iron-replete and iron-restricted conditions, and the downregulation could be reversed by complementation. No significant changes in expression of other iron receptors were detected, suggesting that the MisR/S system specifically regulates hmbR. When hemoglobin was the sole iron source, growth defects were detected in the mutants. Primer extension analysis identified a promoter upstream of the hemO-associated Correia element (CE) and another promoter at the proximal end of CE, and processed transcripts previously identified for other cotranscribed CEs were also detected, suggesting that there may be posttranscriptional regulation. MisR directly interacts with sequences upstream of the CE and upstream of the hmbR Fur binding site and thus independently regulates hemO and hmbR. Analysis of transcriptional reporters of hemO and hmbR further demonstrated the positive role of the MisR/S system and showed that the transcription of hmbR initiated from hemO was significantly reduced. A comparison of the effects of the misS mutation under iron-replete and iron-depleted conditions suggested that activation by the MisR/S system and iron mediated repression by Fur act independently. Thus, the expression of hemO and hmbR is coordinately controlled by multiple independent regulatory mechanisms, including the MisR/S two-component system. PMID- 20008532 TI - Salicylic acid diminishes Staphylococcus aureus capsular polysaccharide type 5 expression. AB - Capsular polysaccharides (CP) of serotypes 5 (CP5) and 8 (CP8) are major Staphylococcus aureus virulence factors. Previous studies have shown that salicylic acid (SAL), the main aspirin metabolite, affects the expression of certain bacterial virulence factors. In the present study, we found that S. aureus strain Reynolds (CP5) cultured with SAL was internalized by MAC-T cells in larger numbers than strain Reynolds organisms not exposed to SAL. Furthermore, the internalization of the isogenic nonencapsulated Reynolds strain into MAC-T cells was not significantly affected by preexposure to SAL. Pretreatment of S. aureus strain Newman with SAL also enhanced internalization into MAC-T cells compared with that of untreated control strains. Using strain Newman organisms, we evaluated the activity of the major cap5 promoter, which was significantly decreased upon preexposure to SAL. Diminished transcription of mgrA and upregulation of the saeRS transcript, both global regulators of CP expression, were found in S. aureus cultured in the presence of SAL, as ascertained by real time PCR analysis. In addition, CP5 production by S. aureus Newman was also decreased by treatment with SAL. Collectively, our data demonstrate that exposure of encapsulated S. aureus strains to low concentrations of SAL reduced CP production, thus unmasking surface adhesins and leading to an increased capacity of staphylococci to invade epithelial cells. The high capacity of internalization of the encapsulated S. aureus strains induced by SAL pretreatment may contribute to the persistence of bacteria in certain hosts. PMID- 20008533 TI - Mapping epitopes of the Plasmodium vivax Duffy binding protein with naturally acquired inhibitory antibodies. AB - Plasmodium vivax Duffy binding protein (DBP) is a merozoite microneme ligand vital for blood-stage infection, which makes it an important candidate vaccine for antibody-mediated immunity against vivax malaria. A differential screen with a linear peptide array compared the reactivities of noninhibitory and inhibitory high-titer human immune sera to identify target epitopes associated with protective immunity. Naturally acquired anti-DBP-specific serologic responses observed in the residents of a region of Papua New Guinea where P. vivax is highly endemic exhibited significant changes in DBP-specific titers over time. The anti-DBP functional inhibition for each serum ranged from complete inhibition to no inhibition even for high-titer responders to the DBP, indicating that epitope specificity is important. Inhibitory immune human antibodies identified specific B-cell linear epitopes on the DBP (SalI) ligand domain that showed significant correlations with inhibitory responses. Affinity-purified naturally acquired antibodies on these epitopes inhibited the DBP erythrocyte binding function greatly, confirming the protective value of specific epitopes. These results represent an important advance in our understanding of part of blood stage immunity to P. vivax and some of the specific targets for vaccine-elicited antibody protection. PMID- 20008534 TI - Klebsiella pneumoniae capsule polysaccharide impedes the expression of beta defensins by airway epithelial cells. AB - Human beta-defensins (hBDs) contribute to the protection of the respiratory tract against pathogens. It is reasonable to postulate that pathogens have developed countermeasures to resist them. Klebsiella pneumoniae capsule polysaccharide (CPS), but not the lipopolysaccharide O antigen, mediated resistance against hBD1 and hBD2. hBD3 was the most potent hBD against Klebsiella. We investigated the possibility that as a strategy for survival in the lung, K. pneumoniae may not activate the expression of hBDs. Infection of A549 and normal human bronchial cells with 52145-Deltawca(K2), a CPS mutant, increased the expression of hBD2 and hBD3. Neither the wild type nor the lipopolysaccharide O antigen mutant increased the expression of hBDs. In vivo, 52145-Deltawca(K2) induced higher levels of mBD4 and mBD14, possible mouse orthologues of hBD2 and hBD3, respectively, than the wild type. 52145-Deltawca(K2)-dependent upregulation of hBD2 occurred via NF kappaB and mitogen-activated protein kinases (MAPKs) p44/42, Jun N-terminal protein kinase (JNK)-dependent pathways. The increase in hBD3 expression was dependent on the MAPK JNK. 52145-Deltawca(K2) engaged Toll-like receptors 2 and 4 (TLR2 and TLR4) to activate hBD2, whereas hBD3 expression was dependent on NOD1. K. pneumoniae induced the expression of CYLD and MKP-1, which act as negative regulators for 52145-Deltawca(K2)-induced expression of hBDs. Bacterial engagement of pattern recognition receptors induced CYLD and MKP-1, which may initiate the attenuation of proinflammatory pathways. The results of this study indicate that K. pneumoniae CPS not only protects the pathogen from the bactericidal action of defensins but also impedes their expression. These features of K. pneumoniae CPS may facilitate pathogen survival in the hostile environment of the lung. PMID- 20008535 TI - Candida glabrata persistence in mice does not depend on host immunosuppression and is unaffected by fungal amino acid auxotrophy. AB - Candida glabrata has emerged as an important fungal pathogen of humans, causing life-threatening infections in immunocompromised patients. In contrast, mice do not develop disease upon systemic challenge, even with high infection doses. In this study we show that leukopenia, but not treatment with corticosteroids, leads to fungal burdens that are transiently increased over those in immunocompetent mice. However, even immunocompetent mice were not capable of clearing infections within 4 weeks. Tissue damage and immune responses to microabscesses were mild as monitored by clinical parameters, including blood enzyme levels, histology, myeloperoxidase, and cytokine levels. Furthermore, we investigated the suitability of amino acid auxotrophic C. glabrata strains for in vitro and in vivo studies of fitness and/or virulence. Histidine, leucine, or tryptophan auxotrophy, as well as a combination of these auxotrophies, did not influence in vitro growth in rich medium. The survival of all auxotrophic strains in immunocompetent mice was similar to that of the parental wild-type strain during the first week of infection and was only mildly reduced 4 weeks after infection, suggesting that C. glabrata is capable of utilizing a broad range of host-derived nutrients during infection. These data suggest that C. glabrata histidine, leucine, or tryptophan auxotrophic strains are suitable for the generation of knockout mutants for in vivo studies. Notably, our work indicates that C. glabrata has successfully developed immune evasion strategies enabling it to survive, disseminate, and persist within mammalian hosts. PMID- 20008536 TI - Toll-like receptor 4 signaling leads to severe fungal infection associated with enhanced proinflammatory immunity and impaired expansion of regulatory T cells. AB - Toll-like receptors (TLRs) present in innate immune cells recognize pathogen molecular patterns and influence immunity to control the host-parasite interaction. The objective of this study was to characterize the involvement of TLR4 in the innate and adaptive immunity to Paracoccidioides brasiliensis, the most important primary fungal pathogen of Latin America. We compared the responses of C3H/HeJ mice, which are naturally defective in TLR4 signaling, with those of C3H/HePas mice, which express functional receptors, after in vitro and in vivo infection with P. brasiliensis. Unexpectedly, we verified that TLR4 defective macrophages infected in vitro with P. brasiliensis presented decreased fungal loads associated with impaired synthesis of nitric oxide, interleukin-12 (IL-12), and macrophage chemotactic protein 1 (MCP-1). After intratracheal infection with 1 million yeasts, TLR4-defective mice developed reduced fungal burdens and decreased levels of pulmonary nitric oxide, proinflammatory cytokines, and antibodies. TLR4-competent mice produced elevated levels of IL-12 and tumor necrosis factor alpha (TNF-alpha), besides cytokines of the Th17 pattern, indicating a proinflammatory role for TLR4 signaling. The more severe infection of TLR4-normal mice resulted in increased influx of activated macrophages and T cells to the lungs and progressive control of fungal burdens but impaired expansion of regulatory T cells (Treg cells). In contrast, TLR4 defective mice were not able to clear their diminished fungal burdens totally, a defect associated with deficient activation of T-cell immunity and enhanced development of Treg cells. These divergent patterns of immunity, however, resulted in equivalent mortality rates, indicating that control of elevated fungal growth mediated by vigorous inflammatory reactions is as deleterious to the hosts as low fungal loads inefficiently controlled by limited inflammatory reactions. PMID- 20008537 TI - Activation of platelets by Aspergillus fumigatus and potential role of platelets in the immunopathogenesis of Aspergillosis. AB - Aspergillus fumigatus is the most frequent cause of invasive mold infections worldwide. Platelets contribute to inflammation and promote thrombosis, characteristically seen in aspergillosis, and might be involved both in antifungal defense and in the histopathological process. In the experiments reported here, in vitro activation of platelets by conidia, swollen conidia, and hyphae from A. fumigatus was assessed by flow cytometry and enzyme immunoassays. THP-1 monocytes and human monocytes with and without platelets were cultured with hyphae from A. fumigatus, and the release of interleukin-8 (IL-8) was measured by enzyme immunoassays. A. fumigatus potently induced the expression of CD62-p and CD63 and the release of CD40 ligand, RANTES, and Dickkopf homolog 1 in platelets, with particularly enhancing effects of hyphae compared with conidia. The hypha mediated activation of platelets further enhanced the release of IL-8 both in THP 1 monocytes and in human adherent monocytes. In conclusion, we have found that A. fumigatus is a potent inducer of platelet-mediated inflammation, potentially promoting protective as well as harmful responses during aspergillosis. PMID- 20008538 TI - Expression of BfrH, a putative siderophore receptor of Bordetella bronchiseptica, is regulated by iron, Fur1, and the extracellular function sigma factor EcfI. AB - Iron (Fe) in soluble elemental form is found in the tissues and fluids of animals at concentrations insufficient for sustaining growth of bacteria. Consequently, to promote colonization and persistence, pathogenic bacteria evolved a myriad of scavenging mechanisms to acquire Fe from the host. Bordetella bronchiseptica, the etiologic agent of upper respiratory infections in a wide range of mammalian hosts, expresses a number of proteins for acquisition of Fe. Using proteomic and genomic approaches, three Fe-regulated genes were identified in the bordetellae: bfrH, a gene encoding a putative siderophore receptor; ecfI, a gene encoding a putative extracellular function (ECF) sigma factor; and ecfR, a gene encoding a putative EcfI modulator. All three genes are highly conserved in B. pertussis, B. parapertussis, and B. avium. Genetic analysis revealed that transcription of bfrH was coregulated by ecfI, ecfR, and fur1, one of two fur homologues carried by B. bronchiseptica. Overexpression of ecfI decoupled bfrH from Fe-dependent regulation. In contrast, expression of bfrH was significantly reduced in an ecfI deletion mutant. Deletion of ecfR, however, was correlated with a significant increase in expression of bfrH, due in part to a cis-acting nucleotide sequence within ecfR which likely reduces the frequency of readthrough transcription of bfrH from the Fe-dependent ecfIR promoter. Using a murine competition infection model, bfrH was shown to be required for optimal virulence of B. bronchiseptica. These experiments revealed ecfIR-bfrH as a locus encoding a new member of the growing family of Fe and ECF sigma factor-modulated regulons in the bordetellae. PMID- 20008539 TI - Shiga toxin 1-induced inflammatory response in lipopolysaccharide-sensitized astrocytes is mediated by endogenous tumor necrosis factor alpha. AB - Hemolytic-uremic syndrome (HUS) is generally caused by Shiga toxin (Stx) producing Escherichia coli. Endothelial dysfunction mediated by Stx is a central aspect in HUS development. However, inflammatory mediators such as bacterial lipopolysaccharide (LPS) and polymorphonuclear neutrophils (PMN) contribute to HUS pathophysiology by potentiating Stx effects. Acute renal failure is the main feature of HUS, but in severe cases, patients can develop neurological complications, which are usually associated with death. Although the mechanisms of neurological damage remain uncertain, alterations of the blood-brain barrier associated with brain endothelial injury is clear. Astrocytes (ASTs) are the most abundant inflammatory cells of the brain that modulate the normal function of brain endothelium and neurons. The aim of this study was to evaluate the effects of Stx type 1 (Stx1) alone or in combination with LPS in ASTs. Although Stx1 induced a weak inflammatory response, pretreatment with LPS sensitized ASTs to Stx1-mediated effects. Moreover, LPS increased the level of expression of the Stx receptor and its internalization. An early inflammatory response, characterized by the release of tumor necrosis factor alpha (TNF-alpha) and nitric oxide and PMN-chemoattractant activity, was induced by Stx1 in LPS-sensitized ASTs, whereas activation, evidenced by higher levels of glial fibrillary acid protein and cell death, was induced later. Furthermore, increased adhesion and PMN-mediated cytotoxicity were observed after Stx1 treatment in LPS-sensitized ASTs. These effects were dependent on NF-kappaB activation or AST-derived TNF-alpha. Our results suggest that TNF-alpha is a pivotal effector molecule that amplifies Stx1 effects on LPS-sensitized ASTs, contributing to brain inflammation and leading to endothelial and neuronal injury. PMID- 20008540 TI - Cigarette smoke exposure impairs pulmonary bacterial clearance and alveolar macrophage complement-mediated phagocytosis of Streptococcus pneumoniae. AB - Cigarette smoke exposure increases the risk of pulmonary and invasive infections caused by Streptococcus pneumoniae, the most commonly isolated organism from patients with community-acquired pneumonia. Despite this association, the mechanisms by which cigarette smoke exposure diminishes host defense against S. pneumoniae infections are poorly understood. In this study, we compared the responses of BALB/c mice following an intratracheal challenge with S. pneumoniae after 5 weeks of exposure to room air or cigarette smoke in a whole-body exposure chamber in vivo and the effects of cigarette smoke on alveolar macrophage phagocytosis of S. pneumoniae in vitro. Bacterial burdens in cigarette smoke exposed mice were increased at 24 and 48 h postinfection, and this was accompanied by a more pronounced clinical appearance of illness, hypothermia, and increased lung homogenate cytokines interleukin-1beta (IL-1beta), IL-6, IL-10, and tumor necrosis factor alpha (TNF-alpha). We also found greater numbers of neutrophils in bronchoalveolar lavage fluid recovered from cigarette smoke exposed mice following a challenge with heat-killed S. pneumoniae. Interestingly, overnight culture of alveolar macrophages with 1% cigarette smoke extract, a level that did not affect alveolar macrophage viability, reduced complement mediated phagocytosis of S. pneumoniae, while the ingestion of unopsonized bacteria or IgG-coated microspheres was not affected. This murine model provides robust additional support to the hypothesis that cigarette smoke exposure increases the risk of pneumococcal pneumonia and defines a novel cellular mechanism to help explain this immunosuppressive effect. PMID- 20008541 TI - High matrix metalloproteinase production correlates with immune activation and leukocyte migration in leprosy reactional lesions. AB - Gelatinases A and B (matrix metalloproteinase 2 [MMP-2] and MMP-9, respectively) can induce basal membrane breakdown and leukocyte migration, but their role in leprosy skin inflammation remains unclear. In this study, we analyzed clinical specimens from leprosy patients taken from stable, untreated skin lesions and during reactional episodes (reversal reaction [RR] and erythema nodosum leprosum [ENL]). The participation of MMPs in disease was suggested by (i) increased MMP mRNA expression levels in skin biopsy specimens correlating with the expression of gamma interferon (IFN-gamma) and tumor necrosis factor alpha (TNF-alpha), (ii) the detection of the MMP protein and enzymatic activity within the inflammatory infiltrate, (iii) increased MMP levels in patient sera, and (iv) the in vitro induction of MMP-9 by Mycobacterium leprae and/or TNF-alpha. It was observed that IFN-gamma, TNF-alpha, MMP-2, and MMP-9 mRNA levels were higher in tuberculoid than lepromatous lesions. In contrast, interleukin-10 and tissue inhibitor of MMP (TIMP-1) message were not differentially modulated. These data correlated with the detection of the MMP protein evidenced by immunohistochemistry and confocal microscopy. When RR and ENL lesions were analyzed, an increase in TNF-alpha, MMP 2, and MMP-9, but not TIMP-1, mRNA levels was observed together with stronger MMP activity (zymography/in situ zymography). Moreover, following in vitro stimulation of peripheral blood cells, M. leprae induced the expression of MMP-9 (mRNA and protein) in cultured cells. Overall, the present data demonstrate an enhanced MMP/TIMP-1 ratio in the inflammatory states of leprosy and point to potential mechanisms for tissue damage. These results pave the way toward the application of new therapeutic interventions for leprosy reactions. PMID- 20008542 TI - Oxidative/nitrosative stress selectively altered A(2B) adenosine receptors in chronic obstructive pulmonary disease. AB - The primary aim of this study was to investigate adenosine receptors (ARs) in bronchoalveolar lavage (BAL) macrophages from patients with chronic obstructive pulmonary disease (COPD) and age-matched healthy smokers. A(2B)ARs were significantly decreased in BAL macrophages from patients with COPD when compared with healthy smokers. The effect of proinflammatory cytokines and oxidative/nitrosative stress on AR expression and function in U937 cells before and after PMA treatment was evaluated. IL-1beta and TNF-alpha treatment up regulated A(2A)- and A(3)ARs but not A(1)- or A(2B)ARs, whereas IL-6 did not modify AR expression. In contrast, oxidative/nitrosative stress selectively decreased A(2B)AR expression, which was associated with a reduction in the potency of the adenosine agonist 5'-N-ethylcarboxamideadenosine (NECA) to induce cAMP. Further, the ability of NECA to enhance cell proliferation was increased after oxidative/nitrosative stress. The specific involvement of A(2B)ARs was investigated by using potent and selective A(2B)AR antagonist and by A(2B)AR knockdown using siRNA and demonstrated responses similar to those obtained with oxidative/nitrosative stress. N-acetylcysteine (NAC), an antioxidant agent, counteracted the decrease in A(2B)AR expression, as well as the altered NECA effects on cAMP and cell proliferation. These findings highlight the central role of A(2B)ARs in alveolar macrophages, suggesting that their modulation could represent an innovative pharmacological strategy to manage COPD.-Varani, K., Caramori, G., Vincenzi, F., Tosi, A., Barczyk, A., Contoli, M., Casolari, P., Triggiani, M., Hansel, T., Leung, E., MacLennan, S., Barnes, P. J., Fan Chung, K., Adcock, I., Papi, A., Borea, P. A. Oxidative/nitrosative stress selectively altered A(2B) adenosine receptors in chronic obstructive pulmonary disease. PMID- 20008543 TI - Suppression of polyglutamine-induced cytotoxicity in Saccharomyces cerevisiae by enhancement of mitochondrial biogenesis. AB - Alterations in mitochondrial metabolism have been associated with age-related neurodegenerative disorders. This is seen in diseases caused by misfolding of proteins with expanded polyglutamine (polyQ) tracts, such as Huntington's disease. Although evidence of mitochondrial impairment has been extensively documented in patients and disease models, the mechanisms involved and their relevance to the initiation of polyQ cytotoxicity and development of clinical manifestations remain controversial. We report that in yeast models of polyQ cytotoxicity, wild-type and mutant polyQ domains might associate early with the outer mitochondrial membrane. The association of mutant domains with mitochondrial membranes could contribute to induce significant changes in mitochondrial physiology, ultimately compromising the cell's ability to respire. The respiratory defect can be fully prevented by enhancing mitochondrial biogenesis by overexpression of Hap4p, the catalytic subunit of the transcriptional activator Hap2/3/4/5p complex, the master regulator of the expression of many nuclear genes encoding mitochondrial proteins in yeast. Protecting cellular respiratory capacity in this way ameliorates the effect of expanded polyQ on cellular fitness. We conclude that mitochondrial dysfunction is an important contributor to polyQ cytotoxicity. Our results suggest that therapeutic approaches enhancing mitochondrial biogenesis could reduce polyQ toxicity and delay the development of clinical symptoms in patients. PMID- 20008544 TI - A VAPB mutant linked to amyotrophic lateral sclerosis generates a novel form of organized smooth endoplasmic reticulum. AB - VAPB (vesicle-associated membrane protein-associated protein B) is an endoplasmic reticulum (ER)-resident tail-anchored adaptor protein involved in lipid transport. A dominantly inherited mutant, P56S-VAPB, causes a familial form of amyotrophic lateral sclerosis (ALS) and forms poorly characterized inclusion bodies in cultured cells. To provide a cell biological basis for the understanding of mutant VAPB pathogenicity, we investigated its biogenesis and the inclusions that it generates. Translocation assays in cell-free systems and in cultured mammalian cells were used to investigate P56S-VAPB membrane insertion, and the inclusions were characterized by confocal imaging and electron microscopy. We found that mutant VAPB inserts post-translationally into ER membranes in a manner indistinguishable from the wild-type protein but that it rapidly clusters to form inclusions that remain continuous with the rest of the ER. Inclusions were induced by the mutant also when it was expressed at levels comparable to the endogenous wild-type protein. Ultrastructural analysis revealed that the inclusions represent a novel form of organized smooth ER (OSER) consisting in a limited number of parallel cisternae (usually 2 or 3) interleaved by a approximately 30 nm-thick electron-dense cytosolic layer. Our results demonstrate that the ALS-linked VAPB mutant causes dramatic ER restructuring that may underlie its pathogenicity in motoneurons. PMID- 20008546 TI - Need for integrative assessment of toxicology data. PMID- 20008545 TI - Tumor growth and angiogenesis are dependent on the presence of immature dendritic cells. AB - Dendritic cells (DCs)--immunomodulatory cells that initiate adaptive immune responses--have recently been shown to exert proangiogenic effects when infiltrating the tumor microenvironment. As tumors that escape immune surveillance inhibit DC maturation, we explored whether maturation status determines their ability to promote angiogenesis and whether angiogenesis depends on the presence of DCs. Using mouse xenograft models of human tumors, we show that fast-growing "angiogenic" tumors are infiltrated by a more immature DC population than respective dormant avascular tumors. Accordingly, supplementation of immature DCs, but not mature DCs, enhanced tumor growth. When DCs were mixed with Matrigel and injected subcutaneously into mice, only immature DCs promoted the ingrowth of patent blood vessels. Notably, depletion of DCs in a transgenic mouse model that allows for their conditional ablation completely abrogated basic fibroblast growth factor-induced angiogenesis in Matrigel plugs, and significantly inhibited tumor growth in these mice. Because immature DCs actively promote angiogenesis and tumor growth, whereas DC maturation or ablation suppresses this response, we conclude that angiogenesis is dependent on the presence of immature DCs. Thus, cancer immunotherapies that promote DC maturation may act by both augmenting the host immune response to the tumor and by suppressing tumor angiogenesis. PMID- 20008547 TI - Corneal neovascularization and ocular irritancy responses in dogs following topical ocular administration of an EP4-prostaglandin E2 agonist. AB - Prostaglandin receptor agonists have intraocular pressure-lowering effects in humans and are of interest in the treatment of glaucoma. The prostanoid receptor agonist PF-04475270 is a potent and selective agonist of the prostaglandin E(2) receptor EP4. This paper characterizes the toxicity associated with topical ocular administration of PF-04475270 in beagles. Dogs were given PF-04475270 topically to the eye on a consecutive daily dosing schedule for one or four weeks followed by a one-or four-week reversal period, respectively. Clinical observations, ophthalmic, and laboratory parameters were recorded. Necropsies were conducted at the end of the dosing and recovery phases, and histologic examinations performed. Corneal neovascularization that was considered adverse was observed at doses of >or=1.0 microg/eye and was not reversed by the end of the recovery phase. Dogs dosed with >or=0.25 microg/eye developed a dose-related conjunctival hyperemia that persisted throughout the reversal period. Corneal neovascular cells stained positive with EP4 and the endothelial biomarker Factor VIII-vWF. Other histopathology findings observed at doses of >or=1.0 microg included single-cell necrosis and neutrophils in the cornea, inflammatory cell infiltrates in the iris/ciliary body, and iridal endothelial cell hypertrophy. A resolving acute to subacute inflammation in the iris/ciliary body was observed after the four-week recovery period. PMID- 20008548 TI - PPAR alpha and PPAR gamma coactivation rapidly induces Egr-1 in the nuclei of the dorsal and ventral urinary bladder and kidney pelvis urothelium of rats. AB - To facilitate studies of the rat bladder carcinogenicity of dual-acting PPAR alpha+gamma agonists, we previously identified the Egr-1 transcription factor as a candidate carcinogenicity biomarker and developed rat models based on coadministration of commercially available specific PPAR alpha and PPAR gamma agonists. Immunohistochemistry for Egr-1 with a rabbit monoclonal antibody demonstrated that male vehicle-treated rats exhibited minimal urothelial expression and specifically, no nuclear signal. In contrast, Egr-1 was induced in the nuclei of bladder, as well as kidney pelvis, urothelia within one day (2 doses) of oral dosing of rats with a combination of 8 mg/kg rosiglitazone and 200 mg/kg fenofibrate (specific PPAR gamma and PPAR alpha agonists, respectively). These findings were confirmed by Western blotting using a different Egr-1 antibody. Egr-1 was induced to similar levels in the dorsal and ventral bladder urothelium, arguing against involvement of urinary solids. Egr-1 induction sometimes occurred in a localized fashion, indicating physiological microheterogeneity in the urothelium. The rapid kinetics supported that Egr-1 induction occurred as a result of pharmacological activation of PPAR alpha and PPAR gamma, which are coexpressed at high levels in the rat urothelium. Finally, our demonstration of a nuclear localization supports that the Egr-1 induced by PPAR alpha and PPAR gamma coactivation in the rat urothelium may be biologically active. PMID- 20008549 TI - A strategy for risk management of drug-induced phospholipidosis. AB - Drug-induced phospholipidosis (PL) is an excessive accumulation of phospholipids and drug in lysosomes. Phospholipidosis signals a change in cell membrane integrity and accumulation of intracellular drug or metabolite in tissues. The sensitivity and susceptibility of preclinical models to detect PL vary with therapeutic agents, and PL is expected to be reversible after discontinuation of drug treatment. The prevailing scientific opinion is that PL by itself is not adverse; however, some regulatory authorities consider PL to be adverse because a small number of chemicals are able to cause PL and concurrent organ toxicity. Until a greater understanding of PL emerges, a well-thought-out risk management strategy for PL will increase confidence in safety and improve selection and development of new drugs. This paper provides a tiered approach to risk management of drug-induced PL. It begins with use of in silico and in vitro tools to design and select compounds with reduced potential to produce PL. Early in vivo studies in two species are used to better characterize potential for toxicity and PL. Finally, routine risk management tools (i.e., translational biomarkers, assessment of reversibility) are used to support confidence in safety of compounds that induce PL in animals. PMID- 20008550 TI - Rap1 in Candida albicans: an unusual structural organization and a critical function in suppressing telomere recombination. AB - Rap1 (repressor activator protein 1) is a conserved multifunctional protein initially identified as a transcriptional regulator of ribosomal protein genes in Saccharomyces cerevisiae but subsequently shown to play diverse functions at multiple chromosomal loci, including telomeres. The function of Rap1 appears to be evolutionarily plastic, especially in the budding yeast lineages. We report here our biochemical and molecular genetic characterizations of Candida albicans Rap1, which exhibits an unusual, miniaturized domain organization in comparison to the S. cerevisiae homologue. We show that in contrast to S. cerevisiae, C. albicans RAP1 is not essential for cell viability but is critical for maintaining normal telomere length and structure. The rap1 null mutant exhibits drastic telomere-length dysregulation and accumulates high levels of telomere circles, which can be largely attributed to aberrant recombination activities at telomeres. Analysis of combination mutants indicates that Rap1 and other telomere proteins mediate overlapping but nonredundant roles in telomere protection. Consistent with the telomere phenotypes of the mutant, C. albicans Rap1 is localized to telomeres in vivo and recognizes the unusual telomere repeat unit with high affinity and sequence specificity in vitro. The DNA-binding Myb domain of C. albicans Rap1 is sufficient to suppress most of the telomere aberrations observed in the null mutant. Notably, we were unable to detect specific binding of C. albicans Rap1 to gene promoters in vivo or in vitro, suggesting that its functions are more circumscribed in this organism. Our findings provide insights on the evolution and mechanistic plasticity of a widely conserved and functionally critical telomere component. PMID- 20008551 TI - A G1 checkpoint mediated by the retinoblastoma protein that is dispensable in terminal differentiation but essential for senescence. AB - Terminally differentiated cell types are needed to live and function in a postmitotic state for a lifetime. Cellular senescence is another type of permanent arrest that blocks the proliferation of cells in response to genotoxic stress. Here we show that the retinoblastoma protein (pRB) uses a mechanism to block DNA replication in senescence that is distinct from its role in permanent cell cycle exit associated with terminal differentiation. Our work demonstrates that a subtle mutation in pRB that cripples its ability to interact with chromatin regulators impairs heterochromatinization and repression of E2F responsive promoters during senescence. In contrast, terminally differentiated nerve and muscle cells bearing the same mutation fully exit the cell cycle and block E2F-responsive gene expression by a different mechanism. Remarkably, this reveals that pRB recruits chromatin regulators primarily to engage a stress responsive G(1) arrest program. PMID- 20008552 TI - The final step in 5.8S rRNA processing is cytoplasmic in Saccharomyces cerevisiae. AB - The 18S rRNA component of yeast (Saccharomyces cerevisiae) 40S ribosomes undergoes cytoplasmic 3' cleavage following nuclear export, whereas exported pre 60S subunits were believed to contain only mature 5.8S and 25S rRNAs. However, in situ hybridization detected 3'-extended forms of 5.8S rRNA in the cytoplasm, which were lost when Crm1-dependent preribosome export was blocked by treatment with leptomycin B (LMB). LMB treatment rapidly blocked processing of 6S pre-rRNA to 5.8S rRNA, leading to TRAMP-dependent pre-rRNA degradation. The 6S pre-rRNA was coprecipitated with the 60S export adapter Nmd3 and cytoplasmic 60S synthesis factor Lsg1. The longer 5.8S+30 pre-rRNA (a form of 5.8S rRNA 3' extended by approximately 30 nucleotides) is processed to 6S by the nuclear exonuclease Rrp6, and nuclear pre-rRNA accumulated in the absence of Rrp6. In contrast, 6S to 5.8S processing requires the cytoplasmic exonuclease Ngl2, and cytoplasmic pre-rRNA accumulated in strains lacking Ngl2. We conclude that nuclear pre-60S particles containing the 6S pre-rRNA bind Nmd3 and Crm1 and are exported to the cytoplasm prior to final maturation by Ngl2. PMID- 20008553 TI - The ubiquitin ligase Hul5 promotes proteasomal processivity. AB - The 26S proteasome is a large cytoplasmic protease that degrades polyubiquitinated proteins to short peptides in a processive manner. The proteasome 19S regulatory subcomplex tethers the target protein via its polyubiquitin adduct and unfolds the target polypeptide, which is then threaded into the proteolytic site-containing 20S subcomplex. Hul5 is a 19S subcomplex associated ubiquitin ligase that elongates ubiquitin chains on proteasome-bound substrates. We isolated hul5 Delta as a mutation with which fusions of an unstable cyclin to stable reporter proteins accumulate as partially processed products. These products appear transiently in the wild type but are strongly stabilized in 19S ATPase mutants and in the hul5 Delta mutant, supporting a role for the ATPase subunits in the unfolding of proteasome substrates before insertion into the catalytic cavity and suggesting a role for Hul5 in the processive degradation of proteins that are stalled on the proteasome. PMID- 20008554 TI - MAGUK-controlled ubiquitination of CARMA1 modulates lymphocyte NF-kappaB activity. AB - The adaptor protein CARMA1 is required for antigen receptor-triggered activation of IKK and JNK in lymphocytes. Once activated, the events that subsequently turn off the CARMA1 signalosome are unknown. In this study, we found that antigen receptor-activated CARMA1 underwent lysine 48 (K48) polyubiquitination and proteasome-dependent degradation. The MAGUK region of CARMA1 was an essential player in this event; the SH3 and GUK domains contained the main ubiquitin acceptor sites, and deletion of a Hook domain (an important structure for maintaining inactive MAGUK proteins) between SH3 and GUK was sufficient to induce constitutive ubiquitination of CARMA1. A similar deletion promoted the ubiquitination of PSD-95 and Dlgh1, suggesting that a conserved mechanism may control the turnover of other MAGUK family protein complexes. Functionally, we demonstrated that elimination of MAGUK ubiquitination sites in CARMA1 resulted in elevated basal and inducible NF-kappaB and JNK activation as a result of defective K48 ubiquitination and increased persistence of this ubiquitination deficient CARMA1 protein in activated lymphocytes. The coordination of degradation with the full activation of the CARMA1 molecule likely provides an intrinsic feedback control mechanism to balance lymphocyte activation upon antigenic stimulation. PMID- 20008555 TI - Pirh2 E3 ubiquitin ligase targets DNA polymerase eta for 20S proteasomal degradation. AB - DNA polymerase eta (PolH), a Y family translesion polymerase, is required for repairing UV-induced DNA damage, and loss of PolH is responsible for early onset of malignant skin cancers in patients with xeroderma pigmentosum variant (XPV), an autosomal recessive disorder. Here, we show that PolH, a target of the p53 tumor suppressor, is a short-half-life protein. We found that PolH is degraded by proteasome, which is enhanced upon UV irradiation. We also found that PolH interacts with Pirh2 E3 ligase, another target of the p53 tumor suppressor, via the polymerase-associated domain in PolH and the RING finger domain in Pirh2. In addition, we show that overexpression of Pirh2 decreases PolH protein stability, whereas knockdown of Pirh2 increases it. Interestingly, we found that PolH is recruited by Pirh2 and degraded by 20S proteasome in a ubiquitin-independent manner. Finally, we observed that Pirh2 knockdown leads to accumulation of PolH and, subsequently, enhances the survival of UV-irradiated cells. We postulate that UV irradiation promotes cancer formation in part by destabilizing PolH via Pirh2-mediated 20S proteasomal degradation. PMID- 20008556 TI - The Caenorhabditis elegans Ste20-related kinase and Rac-type small GTPase regulate the c-Jun N-terminal kinase signaling pathway mediating the stress response. AB - Mitogen-activated protein kinases (MAPKs) are integral to the mechanisms by which cells respond to physiological stimuli and a wide variety of environmental stresses. In Caenorhabditis elegans, the stress response is controlled by a c-Jun N-terminal kinase (JNK)-like MAPK signaling pathway, which is regulated by MLK-1 MAPK kinase kinase (MAPKKK), MEK-1 MAPKK, and KGB-1 JNK-like MAPK. In this study, we identify the max-2 gene encoding a C. elegans Ste20-related protein kinase as a component functioning upstream of the MLK-1-MEK-1-KGB-1 pathway. The max-2 loss of-function mutation is defective in activation of KGB-1, resulting in hypersensitivity to heavy metals. Biochemical analysis reveals that MAX-2 activates MLK-1 through direct phosphorylation of a specific residue in the activation loop of the MLK-1 kinase domain. Our genetic data presented here also show that MIG-2 small GTPase functions upstream of MAX-2 in the KGB-1 pathway. These results suggest that MAX-2 and MIG-2 play a crucial role in mediating the heavy metal stress response regulated by the KGB-1 pathway. PMID- 20008557 TI - Frequency-dependent regulation of follicle-stimulating hormone beta by pulsatile gonadotropin-releasing hormone is mediated by functional antagonism of bZIP transcription factors. AB - Oscillatory synthesis and secretion of the gonadotropins, follicle-stimulating hormone (FSH) and luteinizing hormone (LH), under the control of pulsatile hypothalamic gonadotropin-releasing hormone (GnRH), is essential for normal reproductive development and fertility. The molecular mechanisms by which various patterns of pulsatile GnRH regulate gonadotrope responsiveness remain poorly understood. In contrast to the alpha and LH beta subunit genes, FSH beta subunit transcription is preferentially stimulated at low rather than high frequencies of pulsatile GnRH. In this study, mutation of a cyclic AMP response element (CRE) within the FSH beta promoter resulted in the loss of preferential GnRH stimulation at low pulse frequencies. We hypothesized that high GnRH pulse frequencies might stimulate a transcriptional repressor(s) to attenuate the action of CRE binding protein (CREB) and show that inducible cAMP early repressor (ICER) fulfills such a role. ICER was not detected under basal conditions, but pulsatile GnRH stimulated ICER to a greater extent at high than at low pulse frequencies. ICER binds to the FSH beta CRE site to reduce CREB occupation and abrogates both maximal GnRH stimulation and GnRH pulse frequency-dependent effects on FSH beta transcription. These data suggest that ICER production antagonizes the stimulatory action of CREB to attenuate FSH beta transcription at high GnRH pulse frequencies, thereby playing a critical role in regulating cyclic reproductive function. PMID- 20008558 TI - Rab13 regulates neurite outgrowth in PC12 cells through its effector protein, JRAB/MICAL-L2. AB - Neurite outgrowth is the first step in the processes of neuronal differentiation and regeneration and leads to synaptic polarization and plasticity. Rab13 small G protein shows an increased mRNA expression level during neuronal regeneration; it is therefore thought to be involved in this process. We previously identified JRAB (junctional Rab13-binding protein)/MICAL-L2 (molecules interacting with CasL like 2) as a novel Rab13 effector protein. Here, we show that Rab13 regulates neurite outgrowth in the rat pheochromocytoma cell line PC12 through an interaction with JRAB/MICAL-L2. The expression of JRAB/MICAL-L2 alone inhibits neurite outgrowth, whereas coexpression of the dominant active form of Rab13 rescues this effect. We also demonstrate an intramolecular interaction between the N-terminal calponin-homology (CH) and LIM domains of JRAB/MICAL-L2 and the C terminal coiled-coil domain. Finally, we show that the binding of Rab13 to JRAB/MICAL-L2 stimulates the interaction of JRAB/MICAL-L2 with actinin-4, an actin-binding protein, which localizes to the cell body and the tips of the neurites in PC12 cells. These results suggest that Rab13 and JRAB/MICAL-L2 may act to transfer actinin-4 from the cell body to the tips of neurites, where actinin-4 is involved in the reorganization of the actin cytoskeleton which results in neurite outgrowth. PMID- 20008560 TI - Tarun Kapoor: in the right position to study chromosomes. Interview by Caitlin Sedwick. PMID- 20008561 TI - Cytokinetic astralogy. AB - Division plane specification in animal cells has long been presumed to involve direct contact between microtubules of the anaphase mitotic spindle and the cell cortex. In this issue, von Dassow et al. (von Dassow et al. 2009. J. Cell. Biol. doi:10.1083/jcb.200907090) challenge this assumption by showing that spindle microtubules can effectively position the division plane at a distance from the cell cortex. PMID- 20008562 TI - ISWI is a RanGTP-dependent MAP required for chromosome segregation. AB - Production of RanGTP around chromosomes induces spindle assembly by activating nuclear localization signal (NLS)-containing factors. Here, we show that the NLS protein ISWI, a known chromatin-remodeling ATPase, is a RanGTP-dependent microtubule (MT)-associated protein. Recombinant ISWI induces MT nucleation, stabilization, and bundling in vitro. In Xenopus culture cells and egg extract, ISWI localizes within the nucleus in interphase and on spindles during mitosis. Depletion of ISWI in egg extracts does not affect spindle assembly, but in anaphase spindle MTs disappear and chromosomes do not segregate. We show directly that ISWI is required for the RanGTP-dependent stabilization of MTs during anaphase independently of its effect on chromosomes. ISWI depletion in Drosophila S2 cells induces defects in spindle MTs and chromosome segregation in anaphase, and the cells eventually stop growing. Our results demonstrate that distinctly from its role in spindle assembly, RanGTP maintains spindle MTs in anaphase through the local activation of ISWI and that this is essential for proper chromosome segregation. PMID- 20008563 TI - Action at a distance during cytokinesis. AB - Animal cells decide where to build the cytokinetic apparatus by sensing the position of the mitotic spindle. Reflecting a long-standing presumption that a furrow-inducing stimulus travels from spindle to cortex via microtubules, debate continues about which microtubules, and in what geometry, are essential for accurate cytokinesis. We used live imaging in urchin and frog embryos to evaluate the relationship between microtubule organization and cytokinetic furrow position. In normal cells, the cytokinetic apparatus forms in a region of lower cortical microtubule density. Remarkably, cells depleted of astral microtubules conduct accurate, complete cytokinesis. Conversely, in anucleate cells, asters alone can support furrow induction without a spindle, but only when sufficiently separated. Ablation of a single centrosome displaces furrows away from the remaining centrosome; ablation of both centrosomes causes broad, inefficient furrowing. We conclude that the asters confer accuracy and precision to a primary furrow-inducing signal that can reach the cell surface from the spindle without transport on microtubules. PMID- 20008564 TI - Muscle inactivation of mTOR causes metabolic and dystrophin defects leading to severe myopathy. AB - Mammalian target of rapamycin (mTOR) is a key regulator of cell growth that associates with raptor and rictor to form the mTOR complex 1 (mTORC1) and mTORC2, respectively. Raptor is required for oxidative muscle integrity, whereas rictor is dispensable. In this study, we show that muscle-specific inactivation of mTOR leads to severe myopathy, resulting in premature death. mTOR-deficient muscles display metabolic changes similar to those observed in muscles lacking raptor, including impaired oxidative metabolism, altered mitochondrial regulation, and glycogen accumulation associated with protein kinase B/Akt hyperactivation. In addition, mTOR-deficient muscles exhibit increased basal glucose uptake, whereas whole body glucose homeostasis is essentially maintained. Importantly, loss of mTOR exacerbates the myopathic features in both slow oxidative and fast glycolytic muscles. Moreover, mTOR but not raptor and rictor deficiency leads to reduced muscle dystrophin content. We provide evidence that mTOR controls dystrophin transcription in a cell-autonomous, rapamycin-resistant, and kinase independent manner. Collectively, our results demonstrate that mTOR acts mainly via mTORC1, whereas regulation of dystrophin is raptor and rictor independent. PMID- 20008565 TI - Valosin-containing protein (VCP) is required for autophagy and is disrupted in VCP disease. AB - Mutations in valosin-containing protein (VCP) cause inclusion body myopathy (IBM), Paget's disease of the bone, and frontotemporal dementia (IBMPFD). Patient muscle has degenerating fibers, rimmed vacuoles (RVs), and sarcoplasmic inclusions containing ubiquitin and TDP-43 (TARDNA-binding protein 43). In this study, we find that IBMPFD muscle also accumulates autophagosome-associated proteins, Map1-LC3 (LC3), and p62/sequestosome, which localize to RVs. To test whether VCP participates in autophagy, we silenced VCP or expressed adenosine triphosphatase-inactive VCP. Under basal conditions, loss of VCP activity results in autophagosome accumulation. After autophagic induction, these autophagosomes fail to mature into autolysosomes and degrade LC3. Similarly, IBMPFD mutant VCP expression in cells and animals leads to the accumulation of nondegradative autophagosomes that coalesce at RVs and fail to degrade aggregated proteins. Interestingly, TDP-43 accumulates in the cytosol upon autophagic inhibition, similar to that seen after IBMPFD mutant expression. These data implicate VCP in autophagy and suggest that impaired autophagy explains the pathology seen in IBMPFD muscle, including TDP-43 accumulation. PMID- 20008566 TI - Lipid-regulated sterol transfer between closely apposed membranes by oxysterol binding protein homologues. AB - Sterols are transferred between cellular membranes by vesicular and poorly understood nonvesicular pathways. Oxysterol-binding protein-related proteins (ORPs) have been implicated in sterol sensing and nonvesicular transport. In this study, we show that yeast ORPs use a novel mechanism that allows regulated sterol transfer between closely apposed membranes, such as organelle contact sites. We find that the core lipid-binding domain found in all ORPs can simultaneously bind two membranes. Using Osh4p/Kes1p as a representative ORP, we show that ORPs have at least two membrane-binding surfaces; one near the mouth of the sterol-binding pocket and a distal site that can bind a second membrane. The distal site is required for the protein to function in cells and, remarkably, regulates the rate at which Osh4p extracts and delivers sterols in a phosphoinositide-dependent manner. Together, these findings suggest a new model of how ORPs could sense and regulate the lipid composition of adjacent membranes. PMID- 20008567 TI - Basal body stability and ciliogenesis requires the conserved component Poc1. AB - Centrioles are the foundation for centrosome and cilia formation. The biogenesis of centrioles is initiated by an assembly mechanism that first synthesizes the ninefold symmetrical cartwheel and subsequently leads to a stable cylindrical microtubule scaffold that is capable of withstanding microtubule-based forces generated by centrosomes and cilia. We report that the conserved WD40 repeat domain-containing cartwheel protein Poc1 is required for the structural maintenance of centrioles in Tetrahymena thermophila. Furthermore, human Poc1B is required for primary ciliogenesis, and in zebrafish, DrPoc1B knockdown causes ciliary defects and morphological phenotypes consistent with human ciliopathies. T. thermophila Poc1 exhibits a protein incorporation profile commonly associated with structural centriole components in which the majority of Poc1 is stably incorporated during new centriole assembly. A second dynamic population assembles throughout the cell cycle. Our experiments identify novel roles for Poc1 in centriole stability and ciliogenesis. PMID- 20008568 TI - The dynein regulatory complex is the nexin link and a major regulatory node in cilia and flagella. AB - Cilia and flagella are highly conserved microtubule (MT)-based organelles with motile and sensory functions, and ciliary defects have been linked to several human diseases. The 9 + 2 structure of motile axonemes contains nine MT doublets interconnected by nexin links, which surround a central pair of singlet MTs. Motility is generated by the orchestrated activity of thousands of dynein motors, which drive interdoublet sliding. A key regulator of motor activity is the dynein regulatory complex (DRC), but detailed structural information is lacking. Using cryoelectron tomography of wild-type and mutant axonemes from Chlamydomonas reinhardtii, we visualized the DRC in situ at molecular resolution. We present the three-dimensional structure of the DRC, including a model for its subunit organization and intermolecular connections that establish the DRC as a major regulatory node. We further demonstrate that the DRC is the nexin link, which is thought to be critical for the generation of axonemal bending. PMID- 20008569 TI - Drosophila Raf's N terminus contains a novel conserved region and can contribute to torso RTK signaling. AB - Drosophila Raf (DRaf) contains an extended N terminus, in addition to three conserved regions (CR1-CR3); however, the function(s) of this N-terminal segment remains elusive. In this article, a novel region within Draf's N terminus that is conserved in BRaf proteins of vertebrates was identified and termed conserved region N-terminal (CRN). We show that the N-terminal segment can play a positive role(s) in the Torso receptor tyrosine kinase pathway in vivo, and its contribution to signaling appears to be dependent on the activity of Torso receptor, suggesting this N-terminal segment can function in signal transmission. Circular dichroism analysis indicates that DRaf's N terminus (amino acids 1-117) including CRN (amino acids 19-77) is folded in vitro and has a high content of helical secondary structure as predicted by proteomics tools. In yeast two-hybrid assays, stronger interactions between DRaf's Ras binding domain (RBD) and the small GTPase Ras1, as well as Rap1, were observed when CRN and RBD sequences were linked. Together, our studies suggest that DRaf's extended N terminus may assist in its association with the upstream activators (Ras1 and Rap1) through a CRN mediated mechanism(s) in vivo. PMID- 20008570 TI - A single unpaired and transcriptionally silenced X chromosome locally precludes checkpoint signaling in the Caenorhabditis elegans germ line. AB - In many organisms, female and male meiosis display extensive sexual dimorphism in the temporal meiotic program, the number and location of recombination events, sex chromosome segregation, and checkpoint function. We show here that both meiotic prophase timing and germ-line apoptosis, one output of checkpoint signaling, are dictated by the sex of the germ line (oogenesis vs. spermatogenesis) in Caenorhabditis elegans. During oogenesis in feminized animals (fem-3), a single pair of asynapsed autosomes elicits a checkpoint response, yet an unpaired X chromosome fails to induce checkpoint activation. The single X in males and fem-3 worms is a substrate for the meiotic recombination machinery and repair of the resulting double strand breaks appears to be delayed compared with worms carrying paired X chromosomes. Synaptonemal complex axial HORMA domain proteins, implicated in repair of meiotic double strand breaks (DSBs) and checkpoint function, are assembled and disassembled on the single X similarly to paired chromosomes, but the central region component, SYP-1, is not loaded on the X chromosome in males. In fem-3 worms some X chromosomes achieve nonhomologous self-synapsis; however, germ cells with SYP-1-positive X chromosomes are not preferentially protected from apoptosis. Analyses of chromatin and X-linked gene expression indicate that a single X, unlike asynapsed X chromosomes or autosomes, maintains repressive chromatin marks and remains transcriptionally silenced and suggests that this state locally precludes checkpoint signaling. PMID- 20008571 TI - Genetic control of photoperiod sensitivity in maize revealed by joint multiple population analysis. AB - Variation in maize for response to photoperiod is related to geographical adaptation in the species. Maize possesses homologs of many genes identified as regulators of flowering time in other species, but their relation to the natural variation for photoperiod response in maize is unknown. Candidate gene sequences were mapped in four populations created by crossing two temperate inbred lines to two photoperiod-sensitive tropical inbreds. Whole-genome scans were conducted by high-density genotyping of the populations, which were phenotyped over 3 years in both short- and long-day environments. Joint multiple population analysis identified genomic regions controlling photoperiod responses in flowering time, plant height, and total leaf number. Four key genome regions controlling photoperiod response across populations were identified, referred to as ZmPR1-4. Functional allelic differences within these regions among phenotypically similar founders suggest distinct evolutionary trajectories for photoperiod adaptation in maize. These regions encompass candidate genes CCA/LHY, CONZ1, CRY2, ELF4, GHD7, VGT1, HY1/SE5, TOC1/PRR7/PPD-1, PIF3, ZCN8, and ZCN19. PMID- 20008572 TI - New tools for investigating the comparative biology of Caenorhabditis briggsae and C. elegans. AB - Comparative studies of Caenorhabditis briggsae and C. elegans have provided insights into gene function and developmental control in both organisms. C. elegans is a well developed model organism with a variety of molecular and genetic tools to study gene functions. In contrast, there are only very limited tools available for its closest relative, C. briggsae. To take advantage of the full potential of this comparative approach, we have developed several genetic and molecular tools to facilitate functional analysis in C. briggsae. First, we designed and implemented an SNP-based oligonucleotide microarray for rapid mapping of genetic mutants in C. briggsae. Second, we generated a mutagenized frozen library to permit the isolation of targeted deletions and used the library to recover a deletion mutant of cbr-unc-119 for use as a transgenic marker. Third, we used the cbr-unc-119 mutant in ballistic transformation and generated fluorescently labeled strains that allow automated lineaging and cellular resolution expression analysis. Finally, we demonstrated the potential of automated lineaging by profiling expression of egl-5, hlh-1, and pha-4 at cellular resolution and by detailed phenotyping of the perturbations on the Wnt signaling pathway. These additions to the experimental toolkit for C. briggsae should greatly increase its utility in comparative studies with C. elegans. With the emerging sequence of nematode species more closely related to C. briggsae, these tools may open novel avenues of experimentation in C. briggsae itself. PMID- 20008573 TI - The chromatin-remodeling protein Osa interacts with CyclinE in Drosophila eye imaginal discs. AB - Coordinating cell proliferation and differentiation is essential during organogenesis. In Drosophila, the photoreceptor, pigment, and support cells of the eye are specified in an orchestrated wave as the morphogenetic furrow passes across the eye imaginal disc. Cells anterior of the furrow are not yet differentiated and remain mitotically active, while most cells in the furrow arrest at G(1) and adopt specific ommatidial fates. We used microarray expression analysis to monitor changes in transcription at the furrow and identified genes whose expression correlates with either proliferation or fate specification. Some of these are members of the Polycomb and Trithorax families that encode epigenetic regulators. Osa is one; it associates with components of the Drosophila SWI/SNF chromatin-remodeling complex. Our studies of this Trithorax factor in eye development implicate Osa as a regulator of the cell cycle: Osa overexpression caused a small-eye phenotype, a reduced number of M- and S-phase cells in eye imaginal discs, and a delay in morphogenetic furrow progression. In addition, we present evidence that Osa interacts genetically and biochemically with CyclinE. Our results suggest a dual mechanism of Osa function in transcriptional regulation and cell cycle control. PMID- 20008574 TI - Regulation of Salmonella enterica pathogenicity island 1 by DNA adenine methylation. AB - DNA adenine methylase (Dam(-)) mutants of Salmonella enterica are attenuated in the mouse model and present multiple virulence-related defects. Impaired interaction of Salmonella Dam(-) mutants with the intestinal epithelium has been tentatively correlated with reduced secretion of pathogenicity island 1 (SPI-1) effectors. In this study, we show that S. enterica Dam(-) mutants contain lowered levels of the SPI-1 transcriptional regulators HilA, HilC, HilD, and InvF. Epistasis analysis indicates that Dam-dependent regulation of SPI-1 requires HilD, while HilA, HilC, and InvF are dispensable. A transcriptional hilDlac fusion is expressed at similar levels in Dam(+) and Dam(-) hosts. However, lower levels of hilD mRNA are found in a Dam(-) background, thus providing unsuspected evidence that Dam methylation might exert post-transcriptional regulation of hilD expression. This hypothesis is supported by the following lines of evidence: (i) lowered levels of hilD mRNA are found in Salmonella Dam(-) mutants when hilD is transcribed from a heterologous promoter; (ii) increased hilD mRNA turnover is observed in Dam(-) mutants; (iii) lack of the Hfq RNA chaperone enhances hilD mRNA instability in Dam(-) mutants; and (iv) lack of the RNA degradosome components polynucleotide phosphorylase and ribonuclease E suppresses hilD mRNA instability in a Dam(-) background. Our report of Dam-dependent control of hilD mRNA stability suggests that DNA adenine methylation plays hitherto unknown roles in post-transcriptional control of gene expression. PMID- 20008575 TI - A hidden markov model combining linkage and linkage disequilibrium information for haplotype reconstruction and quantitative trait locus fine mapping. AB - Faithful reconstruction of haplotypes from diploid marker data (phasing) is important for many kinds of genetic analyses, including mapping of trait loci, prediction of genomic breeding values, and identification of signatures of selection. In human genetics, phasing most often exploits population information (linkage disequilibrium), while in animal genetics the primary source of information is familial (Mendelian segregation and linkage). We herein develop and evaluate a method that simultaneously exploits both sources of information. It builds on hidden Markov models that were initially developed to exploit population information only. We demonstrate that the approach improves the accuracy of allele phasing as well as imputation of missing genotypes. Reconstructed haplotypes are assigned to hidden states that are shown to correspond to clusters of genealogically related chromosomes. We show that these cluster states can directly be used to fine map QTL. The method is computationally effective at handling large data sets based on high-density SNP panels. PMID- 20008578 TI - Rebuttal: Should prescribing authority be shared with nonphysicians?: yes. PMID- 20008581 TI - Collaboration between family physicians and community pharmacists to enhance adherence to chronic medications: opinions of Saskatchewan family physicians. AB - OBJECTIVE: To ascertain the opinions of family physicians about medication adherence in patients with chronic diseases and the role of community pharmacists in improving adherence to chronic medications, as well as their opinions on increased collaboration with pharmacists to enhance medication adherence. DESIGN: A self-administered postal survey of 19 questions, with opinions collected by ordinal (5-point Likert scale) and open responses. SETTING: Saskatchewan. PARTICIPANTS: Two hundred and eighty-six family physicians working in Saskatchewan in January 2008. MAIN OUTCOME MEASURES: Descriptive statistics of physicians' opinions on the following: medication adherence in patients with chronic diseases; their current interaction with community pharmacists; and potential collaborative strategies to promote medication adherence. RESULTS: The response rate was 39.4%. Approximately 75% of the physicians acknowledged that nonadherence to chronic medications was a problem among their patients. Medication costs and side effects were identified as the 2 most common reasons for medication nonadherence. Only one-quarter of physicians communicated regularly with community pharmacists about adherence issues; most of these physicians were rural physicians. Most physicians agreed that increased collaboration with pharmacists would improve adherence, although support for potential interactions with pharmacists varied. Concerns were expressed about time required by physicians and financial reimbursement. Physicians in practice for less than 10 years and those practising in rural areas were more willing to share clinical information and communicate with pharmacists to promote medication adherence. CONCLUSION: Saskatchewan family physicians appreciate the importance of medication nonadherence but currently seldom interact with community pharmacists on this issue. They believe that pharmacists have a role in supporting patients with medication adherence and indicate a willingness to work more collaboratively with them to promote adherence. For this type of collaboration to be effective, it appears that increased adherence-related communication between the 2 health care providers and additional health care funding are required. PMID- 20008582 TI - Randomized controlled trial of anticipatory and preventive multidisciplinary team care: for complex patients in a community-based primary care setting. AB - OBJECTIVE: T o examine whether quality of care (QOC) improves when nurse practitioners and pharmacists work with family physicians in community practice and focus their work on patients who are 50 years of age and older and considered to be at risk of experiencing adverse health outcomes. DESIGN: Randomized controlled trial. SETTING: A family health network with 8 family physicians, 5 nurses, and 11 administrative personnel serving 10 000 patients in a rural area near Ottawa, Ont. PARTICIPANTS: Patients 50 years of age and older at risk of experiencing adverse health outcomes (N = 241). INTERVENTIONS: At-risk patients were randomly assigned to receive usual care from their family physicians or Anticipatory and Preventive Team Care (APTCare) from a collaborative team composed of their physicians, 1 of 3 nurse practitioners, and a pharmacist. MAIN OUTCOME MEASURES: Quality of care for chronic disease management (CDM) for diabetes, coronary artery disease, congestive heart failure, and chronic obstructive pulmonary disease. RESULTS: Controlling for baseline demographic characteristics, the APTCare approach improved CDM QOC by 9.2% (P < .001) compared with traditional care. The APTCare intervention also improved preventive care by 16.5% (P < .001). We did not observe significant differences in other secondary outcome measures (intermediate clinical outcomes, quality of life [Short-Form 36 and health-related quality of life scales], functional status [instrumental activities of daily living scale] and service usage). CONCLUSION: Additional resources in the form of collaborative multidisciplinary care teams with intensive interventions in primary care can improve QOC for CDM in a population of older at-risk patients. The appropriateness of this intervention will depend on its cost-effectiveness. TRIAL REGISTRATION NUMBER NCT00238836 (CONSORT). PMID- 20008583 TI - Pharmacist and physician collaborative prescribing: for medication renewals within a primary health centre. AB - OBJECTIVE: To determine if there is improvement in medication management when pharmacists and family physicians collaborate to prescribe medication renewals requested by fax. DESIGN: Prospective, non-randomized controlled trial. SETTING: W est Winds Primary Health Centre, an interdisciplinary health centre that includes an academic family medicine practice, located in Saskatoon, Sask. PARTICIPANTS: All patients whose pharmacies faxed the health centre requesting prescription renewals between October 2007 and February 2008 were selected to participate in the study. INTERVENTIONS: Medication renewal requests were forwarded to the pharmacist (who works in the clinic part-time) on days when he was working (intervention group). The pharmacist assessed drug-therapy issues that might preclude safe and effective prescribing of the medication. The pharmacist and physician then made a collaborative decision to authorize the requested medication or to request additional interventions first (eg, perform laboratory tests). When the pharmacist was not working, the physicians managed the renewal requests independently (control group). MAIN OUTCOME MEASURES: Medication renewals authorized with no recommendations, medication-related problems identified, new monitoring tests ordered, and new appointments scheduled with health providers. RESULTS: A total of 181 renewal requests were included (94 in the control group and 87 in the intervention group). The control group had significantly more requests authorized with no recommendations (75.5% vs 52.9%, P = .001). Those in the intervention group had significantly more medication related problems identified (26 vs 10, P = .031); medication changes made (24 vs 10, P = .044); and new appointments scheduled with their family physicians (31 vs 21, P = .049). CONCLUSION: There is an improvement in medication management when a pharmacist collaborates with family physicians to prescribe medication renewals. The collaborative model created significantly more activity with each renewal request (ie, identification of medication-related problems, medication changes, and new appointments), which reflects an improvement in the process of care. PMID- 20008585 TI - For family physicians at the heart of interprofessionalism. PMID- 20008584 TI - Genetic education for primary care providers: improving attitudes, knowledge, and confidence. AB - OBJECTIVE: To increase primary care providers' awareness and use of genetic services; increase their knowledge of genetic issues; increase their confidence in core genetic competencies; change their attitudes toward genetic testing for hereditary diseases; and increase their confidence as primary care genetic resources. DESIGN: Participants completed a workshop and 3 questionnaires: a baseline questionnaire, a survey that provided immediate feedback on the workshop itself, and a follow-up questionnaire 6 months later. SETTING: Ontario. PARTICIPANTS: Primary care providers suggested by deans of nursing, midwifery, family medicine, and obstetric programs, as well as coordinators of nurse practitioner programs, in Ontario and by the Ontario College of Family Physicians. INTERVENTION: A complex educational intervention was developed, including an interactive workshop and PowerPoint educational modules on genetic topics for participants' use (available at www.mtsinai.on.ca/FamMedGen/). MAIN OUTCOME MEASURES: Awareness and use of genetic services, knowledge of genetics, confidence in core clinical genetic skills, attitudes toward genetic testing, and teaching activities related to genetics. RESULTS: The workshop was attended by 29 participants; of those, 21 completed the baseline questionnaire and the 6-month follow-up questionnaire. There was no significant change found in awareness or reported use of genetic services. There was significant improvement in self assessed knowledge of (P = .001) and confidence in (P = .005) skills related to adult-onset genetic disorders. There were significant increases in confidence in many core genetic competencies, including assessing risk of hereditary disorders (P = .033), deciding who should be offered referral for genetic counseling (P = .003), discussing prenatal testing options (P = .034), discussing benefits, risks, and limitations of genetic testing (P = .033), and describing what to expect at a genetic counseling session (P = .022). There was a significant increase in the number of primary care providers agreeing that genetic testing was beneficial in the management of adult-onset diseases (P = .031) and in their confidence in being primary care genetic resources for adult-onset genetic disorders (P = .006). CONCLUSION: Educational interventions that include interactive peer resource workshops and educational modules can increase knowledge of and confidence in the core competencies needed for the delivery of genetic services in primary care. PMID- 20008587 TI - The way we do things around here: advancing an interprofessional care culture within primary care. PMID- 20008588 TI - Should prescribing authority be shared with nonphysicians?: yes. PMID- 20008589 TI - Should prescribing authority be shared with nonphysicians?: no. PMID- 20008590 TI - Medical profession should not be coerced by ideologies. PMID- 20008591 TI - Experience and expertise over evidence. PMID- 20008592 TI - Belling the cat. PMID- 20008593 TI - In agreement. PMID- 20008594 TI - Professionalism in vogue. PMID- 20008595 TI - Early diagnosis of neonatal cholestatic jaundice: test at 2 weeks. AB - OBJECTIVE: To review best practices for early recognition and treatment of conditions resulting in neonatal cholestasis, in order to improve long-term outcomes for affected infants. QUALITY OF EVIDENCE: Studies, review articles, and meta-analyses pertaining to neonatal-onset cholestasis were sought via electronic databases. Reference lists of studies and review articles supplemented the electronic search. Studies were included if they examined the importance of early diagnosis and intervention for cholestatic jaundice of any cause, and mainly comprised Level II and Level III evidence. MAIN MESSAGE: Review of the relevant literature supports the recommendation that infants with jaundice at 2 weeks of age should be tested for cholestasis by quantifying the direct reacting bilirubin levels in their blood. Subsequent rapid investigation using a diagnostic algorithm enables early diagnosis of the specific cause and facilitates timely intervention for conditions whose outcomes are improved by early treatment. CONCLUSION: Universal screening for neonatal cholestasis might help with early identification of cases and improve outcomes, although further study is required in the North American setting. PMID- 20008596 TI - Exposure to fifth disease in pregnancy. AB - QUESTION: One of my pregnant patients came for a routine prenatal visit at 20 weeks' gestation. Near the end of the consultation, she asked me about "slapped cheek" disease and pregnancy, as her son had been diagnosed with fifth disease the previous week. What is the current guideline for pregnant women exposed to parvovirus B19? ANSWER: The rate of vertical transmission during maternal parvovirus B19 infection is estimated at 33%, with fetal complications occurring in 3% of infected women. Fetal complications comprising hemolysis, anemia, and nonimmune hydrops fetalis and fetal loss are more frequent when maternal infection occurs before 20 weeks of gestation. The first step in the management of this patient would be to obtain immunoglobulin (Ig) M and IgG titres against parvovirus to evaluate if the patient has had previous immunity against the disease. If results are negative for IgG but positive for IgM (ie, primary infection), this patient would need close obstetrical monitoring for the following weeks, including serial ultrasounds to rule out fetal anemia and hydrops fetalis. PMID- 20008597 TI - Use of oseltamivir in children. AB - QUESTION: Because of the recent outbreak of pandemic H1N1 2009, I am anticipating a large number of children with influenza-like symptoms or children diagnosed with influenza. Is oseltamivir effective and safe when used for children? ANSWER: Oseltamivir is effective for prevention of complications associated with influenza A (including H1N1) in children. Oseltamivir also reduces the duration of influenza by a median of 36 hours, with nausea and vomiting as the primary reported adverse effects. The World Health Organization recommends oseltamivir as first-line treatment for H1N1, with the use of zanamivir only for suspected or confirmed oseltamivir resistance. Recently, in preparation for the influenza A (H1N1) 2009 pandemic, Health Canada published an interim order permitting the expanded use of oseltamivir for treatment or prophylaxis for children younger than 1 year of age. PMID- 20008598 TI - How long have I got? PMID- 20008599 TI - Dermacase. Can you identify this condition? Nail lichen planus. PMID- 20008600 TI - Genetics: schizophrenia. PMID- 20008602 TI - Handling cues from patients. PMID- 20008601 TI - Taking the stress out of managing gout. PMID- 20008603 TI - Collaboration with other professionals to provide patient care. PMID- 20008604 TI - Mechanisms for communicating within primary health care teams. AB - OBJECTIVE: To explore the types of communication used within primary health care teams (PHCTs), with a particular focus on the mechanisms teams use to promote optimal clinical and administrative information sharing. DESIGN: A descriptive qualitative study. SETTING: Primary health care teams in Ontario between August 2004 and October 2005. PARTICIPANTS: Purposive sampling was used to recruit 121 members from 16 PHCTs reflecting a range of health care professionals, including family physicians, nurse practitioners, nurses, pharmacists, dietitians, social workers, office managers, health promoters, and receptionists. METHODS: Individual in-depth interviews were conducted. An iterative analysis process was used to examine the verbatim transcripts created from the interviews. Techniques of immersion and crystallization were used in the analysis. MAIN FINDINGS: Analysis of the data revealed that communication occurs through formal and informal means. Formal communication included regular team meetings with agendas and meeting minutes, memorandums, computer-assisted communication, and communication logs. Informal communication methods were open and opportunistic, reflecting the traditional hallway consultation. For patient care issues, face-to face communication was preferred. Team member attributes facilitating communication included approachability, availability, and proximity. Finally, funding issues could be an impediment to optimal communication. CONCLUSION: Primary health care is experiencing demands for enhanced and efficient communication that optimizes team functioning and patient care. This study describes formal and informal mechanisms of communication currently used by PHCTs. Attributes that facilitate team communication, such as approachability, availability, and proximity of team members, were highlighted. New funding arrangements might alleviate concerns about remuneration for attendance at meetings. PMID- 20008605 TI - Demands, values, and burnout: relevance for physicians. AB - OBJECTIVE: T o explore the interaction between workload and values congruence (personal values with health care system values) in the context of burnout and physician engagement and to explore the relative importance of these factors by sex, given the distinct work patterns of male and female physicians. DESIGN: National mailed survey. SETTING: Canada. PARTICIPANTS: A random sample of 8100 Canadian physicians (response rate 40%, N = 3213); 2536 responses (from physicians working more than 35 hours per week) were analyzed. MAIN OUTCOME MEASURES: Levels of burnout, values congruence, and workload, by sex, measured by the Maslach Burnout Inventory-General Scale and the Areas of Worklife Scale. RESULTS: Results showed a moderate level of burnout among Canadian physicians, with relatively positive scores on exhaustion, average scores on cynicism, and mildly negative scores on professional efficacy. A series of multiple regression analyses confirmed parallel main effect contributions from manageable workload and values congruence. Both workload and values congruence predicted exhaustion and cynicism for men and women (P = .001). Only values congruence provided a significant prediction of professional efficacy for both men and women (P = .001) These predictors interacted for women on all 3 aspects of burnout (exhaustion, cynicism, and diminished efficacy). Howevever, overall levels of the burnout indicators departed only modestly from normative levels. CONCLUSION: W orkload and values congruence make distinct contributions to physician burnout. Work overload contributes to predicting exhaustion and cynicism; professional values crises contribute to predicting exhaustion, cynicism, and low professional efficacy. The interaction of values and workload for women in particular has implications for the distinct work-life patterns of male and female physicians. Specifically, the congruence of individual values with values inherent in the health care system appeared to be of greater consequence for women than for men. PMID- 20008606 TI - Field notes: assisting achievement and documenting competence. PMID- 20008611 TI - The breast cancer alternative hypothesis: is there evidence to justify replacing it? PMID- 20008612 TI - Chronic myeloid leukemia: reversing the chronic phase. PMID- 20008614 TI - Changes in PSA kinetics after DNA vaccine therapy-not so fast! PMID- 20008617 TI - Findings that hospital factors are surrogate for disparities are misleading. PMID- 20008618 TI - Antitumor effect of somatostatin analogs in neuroendocrine tumors. PMID- 20008619 TI - Novel influenza A (H1N1) in patients with hematologic malignancies. PMID- 20008620 TI - Results of dasatinib therapy in patients with early chronic-phase chronic myeloid leukemia. AB - PURPOSE: Dasatinib is effective therapy for chronic myeloid leukemia (CML) after imatinib failure. In this study, we investigate the efficacy of dasatinib as initial therapy for patients with CML in early chronic phase. PATIENTS AND METHODS: Patients with newly diagnosed CML in early chronic phase were randomly assigned to receive dasatinib 100 mg once daily or 50 mg twice daily as initial therapy. RESULTS: Among 50 patients observed for at least 3 months, 49 patients (98%) achieved a complete cytogenetic response (CCyR), and 41 patients (82%) achieved a major molecular response (MMR). Responses occurred rapidly, with 94% of patients achieving CCyR by 6 months. There was no difference in response rate by treatment arm. The projected event-free survival rate at 24 months is 88%, and all patients are alive after a median follow-up time of 24 months. Grade >or= 3 neutropenia and thrombocytopenia occurred in 21% and 10% of patients, respectively. Nonhematologic toxicity was usually grade 1 to 2. There was no significant difference in toxicity between the two arms, and the actual median dose at 12 months was 100 mg (range, 20 to 100 mg). CONCLUSION: Dasatinib is an effective agent for the initial management of CML in early chronic phase, producing high rates of CCyR and MMR. PMID- 20008621 TI - Nilotinib as front-line treatment for patients with chronic myeloid leukemia in early chronic phase. AB - PURPOSE: Although most patients with chronic myeloid leukemia (CML) in chronic phase respond well to front-line therapy with imatinib, some patients do not achieve the desirable end point, and others may eventually lose response or are intolerant. PATIENTS AND METHODS: Patients with newly diagnosed CML in chronic phase were treated with nilotinib 400 mg twice daily on an empty stomach as initial therapy. RESULTS: Among 51 patients in chronic phase observed for at least 3 months, 50 (98%) achieved a complete cytogenetic remission (CCyR), and 39 (76%) achieved a major molecular response (MMR). Responses occurred rapidly, with 96% of patients achieving CCyR by 3 months and 98% achieving CCyR by 6 months. The projected event-free survival at 24 months is 90%, and all patients are alive after a median follow-up time of 17 months. Grade >or= 3 neutropenia occurred in 12% of patients, and thrombocytopenia in occurred 11%. Nonhematologic toxicity was usually grade 1 to 2 and manageable. The actual median dose at 12 months was 800 mg (range, 200 to 800 mg). CONCLUSION: Nilotinib is an effective option for the initial management of CML in early chronic phase, producing high rates of CCyR and MMR, with most patients reaching these responses early during their therapy. PMID- 20008622 TI - Phase III, randomized, open-label study of daily imatinib mesylate 400 mg versus 800 mg in patients with newly diagnosed, previously untreated chronic myeloid leukemia in chronic phase using molecular end points: tyrosine kinase inhibitor optimization and selectivity study. AB - PURPOSE: To evaluate the safety and efficacy of initial treatment with imatinib mesylate 800 mg/d (400 mg twice daily) versus 400 mg/d in patients with newly diagnosed chronic myeloid leukemia in chronic phase. PATIENTS AND METHODS: A total of 476 patients were randomly assigned 2:1 to imatinib 800 mg (n = 319) or 400 mg (n = 157) daily. The primary end point was the major molecular response (MMR) rate at 12 months. RESULTS: At 12 months, differences in MMR and complete cytogenetic response (CCyR) rates were not statistically significant (MMR, 46% v 40%; P = .2035; CCyR, 70% v 66%; P = .3470). However, MMR occurred faster among patients randomly assigned to imatinib 800 mg/d, who had higher rates of MMR at 3 and 6 months compared with those in the imatinib 400-mg/d arm (P = .0035 by log rank test). CCyR also occurred faster in the 800-mg/d arm (CCyR at 6 months, 57% v 45%; P = .0146). The most common adverse events were edema, gastrointestinal problems, and rash, and all were more common in patients in the 800-mg/d arm. Grades 3 to 4 hematologic toxicity also occurred more frequently in patients receiving imatinib 800 mg/d. CONCLUSION: MMR rates at 1 year were similar with imatinib 800 mg/d and 400 mg/d, but MMR and CCyR occurred earlier in patients treated with 800 mg/d. Continued follow-up is needed to determine the clinical significance of earlier responses on high-dose imatinib. PMID- 20008623 TI - Screening for founder mutations in BRCA1 and BRCA2 in unselected Jewish women. AB - PURPOSE: There are two mutations in BRCA1 and one mutation in BRCA2 that are present in up to 2.5% of Ashkenazi Jewish women. Current guidelines for testing stipulate that a personal or family history of cancer be present to be eligible for testing. To date, population screening in this population has not been suggested. However, this may be rational. Little is known about the appropriateness of testing guidelines for the Jewish population or the level of interest in testing. METHODS: Eligible subjects were women who self-identified as Jewish, who were between the ages of 25 and 80 years, and who resided in Ontario. Subjects were recruited through an article in a national newspaper. Women were asked to complete a study questionnaire and a family history questionnaire and to provide a blood or saliva sample. The risk of carrying a BRCA mutation was estimated for each woman. Results A total of 2,080 women were enrolled onto the study. The overall mutation prevalence was 1.1% (0.5% for BRCA1 and 0.6% for BRCA2). Among the 22 mutation carriers, the mean estimate of carrying a BRCA mutation was 3.9%. Ten (45%) of the 22 women met the current Ontario Ministry of Health Guidelines criteria for testing. CONCLUSION: There is considerable interest for genetic testing among Jewish women at low risk of carrying a mutation. However, many women with mutations are ineligible for genetic testing under current guidelines. Approximately 1% of Jewish women carry a BRCA mutation, and these women should be considered to be candidates for genetic testing. PMID- 20008624 TI - Phase II trial of infusional fluorouracil, irinotecan, and bevacizumab for metastatic colorectal cancer: efficacy and circulating angiogenic biomarkers associated with therapeutic resistance. AB - PURPOSE: We investigated the efficacy of fluorouracil (FU), leucovorin, irinotecan, and bevacizumab (FOLFIRI + B) in a phase II trial in patients previously untreated for metastatic colorectal cancer (mCRC), and changes during treatment in plasma cytokines and angiogenic factors (CAFs) as potential markers of treatment response and therapeutic resistance. PATIENTS AND METHODS: We conducted a phase II, two-institution trial of FOLFIRI + B. Each 14-day cycle consisted of bevacizumab (5 mg/kg), irinotecan (180 mg/m(2)), bolus FU (400 mg/m(2)), and leucovorin (400 mg/m(2)) followed by a 46-hour infusion of FU (2,400 mg/m(2)). Levels of 37 CAFs were assessed using multiplex-bead assays and enzyme-linked immunosorbent assay at baseline, during treatment, and at the time of progressive disease (PD). RESULTS: Forty-three patients were enrolled. Median progression-free survival (PFS), the primary end point of the study, was 12.8 months. Median overall survival was 31.3 months, with a response rate of 65%. Elevated interleukin-8 at baseline was associated with a shorter PFS (11 v 15.1 months, P = .03). Before the radiographic development of PD, several CAFs associated with angiogenesis and myeloid recruitment increased compared to baseline, including basic fibroblast growth factor (P = .046), hepatocyte growth factor (P = .046), placental growth factor (P < .001), stromal-derived factor-1 (P = .04), and macrophage chemoattractant protein-3 (P < .001). CONCLUSION: Efficacy and tolerability of FOLFIRI + B appeared favorable to historical controls in this single arm study. Before radiographic progression, there was a shift in balance of CAFs, with a rise in alternate pro-angiogenic cytokines and myeloid recruitment factors in subsets of patients that may represent mechanisms of resistance. PMID- 20008625 TI - Provision of spiritual care to patients with advanced cancer: associations with medical care and quality of life near death. AB - PURPOSE: To determine whether spiritual care from the medical team impacts medical care received and quality of life (QoL) at the end of life (EoL) and to examine these relationships according to patient religious coping. PATIENTS AND METHODS: Prospective, multisite study of patients with advanced cancer from September 2002 through August 2008. We interviewed 343 patients at baseline and observed them (median, 116 days) until death. Spiritual care was defined by patient-rated support of spiritual needs by the medical team and receipt of pastoral care services. The Brief Religious Coping Scale (RCOPE) assessed positive religious coping. EoL outcomes included patient QoL and receipt of hospice and any aggressive care (eg, resuscitation). Analyses were adjusted for potential confounders and repeated according to median-split religious coping. RESULTS: Patients whose spiritual needs were largely or completely supported by the medical team received more hospice care in comparison with those not supported (adjusted odds ratio [AOR] = 3.53; 95% CI, 1.53 to 8.12, P = .003). High religious coping patients whose spiritual needs were largely or completely supported were more likely to receive hospice (AOR = 4.93; 95% CI, 1.64 to 14.80; P = .004) and less likely to receive aggressive care (AOR = 0.18; 95% CI, 0.04 to 0.79; P = .02) in comparison with those not supported. Spiritual support from the medical team and pastoral care visits were associated with higher QOL scores near death (20.0 [95% CI, 18.9 to 21.1] v 17.3 [95% CI, 15.9 to 18.8], P = .007; and 20.4 [95% CI, 19.2 to 21.1] v 17.7 [95% CI, 16.5 to 18.9], P = .003, respectively). CONCLUSION: Support of terminally ill patients' spiritual needs by the medical team is associated with greater hospice utilization and, among high religious copers, less aggressive care at EoL. Spiritual care is associated with better patient QoL near death. PMID- 20008626 TI - Safety and efficacy of romiplostim in patients with lower-risk myelodysplastic syndrome and thrombocytopenia. AB - PURPOSE: To assess the safety and efficacy of romiplostim, a peptibody that increases platelet production, for treatment of thrombocytopenic patients with myelodysplastic syndromes (MDS). PATIENTS AND METHODS: Eligible patients had lower-risk MDS (International Prognostic Scoring System low or intermediate 1), a mean baseline platelet count or= 65 years at date of diagnosis, and permission from attending physician to contact. Data were collected over 7 years of follow up from consenting patients' medical records, telephone interviews, physician questionnaires, and the National Death Index. Outcomes included self-reported treatment tolerance and all-cause mortality. Four GA domains were described by six individual measures, as follows: sociodemographic by adequate finances; clinical by Charlson comorbidity index (CCI) and body mass index; function by number of physical function limitations; and psychosocial by the five-item Mental Health Index (MHI5) and Medical Outcomes Study Social Support Survey (MOS-SSS). Associations were evaluated using t tests, chi(2) tests, and regression analyses. RESULTS: In multivariable regression including age and stage, three measures from two domains (clinical and psychosocial) were associated with poor treatment tolerance; these were CCI >or= 1 (odds ratio [OR] = 2.49; 95% CI, 1.18 to 5.25), MHI5 score less than 80 (OR = 2.36; 95% CI, 1.15 to 4.86), and MOS-SSS score less than 80 (OR = 3.32; 95% CI, 1.44 to 7.66). Four measures representing all four GA domains predicted mortality; these were inadequate finances (hazard ratio [HR] = 1.89; 95% CI, 1.24 to 2.88; CCI >or= 1 (HR = 1.38; 95% CI, 1.01 to 1.88), functional limitation (HR = 1.40; 95% CI, 1.01 to 1.93), and MHI5 score less than 80 (HR = 1.34; 95% CI, 1.01 to 1.85). In addition, the proportion of women with these outcomes incrementally increased as the number of GA deficits increased. CONCLUSION: This study provides longitudinal evidence that GA domains are associated with poor treatment tolerance and predict mortality at 7 years of follow-up, independent of age and stage of disease. PMID- 20008638 TI - Statin use and prognosis in patients with diffuse large B-cell lymphoma and follicular lymphoma in the rituximab era. AB - PURPOSE: Statins have antilymphoma properties but have also been shown to inhibit the binding of rituximab to the CD20 antigen, resulting in reduced antitumor activity of rituximab in vitro. The clinical impact of statin use on the outcome of lymphoma patients treated with a rituximab-containing regimen is unknown. PATIENTS AND METHODS: Consecutive patients with newly diagnosed, diffuse large B cell lymphoma (DLBCL) and follicular lymphoma (FL) were enrolled onto a registry and observed prospectively. The impact of statin use on patients' outcomes was analyzed. RESULTS: Two hundred twenty-eight patients with DLBCL and 293 patients with FL were enrolled from September 2002 through June 2007; 21% of patients with DLBCL and 19% of patients with FL were on statins at diagnosis, and 20% and 17% remained on statins during lymphoma treatment, respectively. All patients with DLBCL and 39% of patients with FL received initial therapy containing rituximab. The median follow-up time was 47 months (range, 13 to 80 months). Statin use had no impact on the overall response rate (P = .67), overall survival (P = .76), or event-free survival (EFS) in patients with DLBCL (hazard ratio [HR] = 0.85; 95% CI, 0.43 to 1.68). Statin use at diagnosis was associated with improved EFS in patients with FL (HR = 0.45; 95% CI, 0.26 to 0.77), including subgroups treated with rituximab or a rituximab-containing regimen (HR = 0.38; 95% CI, 0.14 to 1.07) and patients who were observed only (HR = 0.38; 95% CI, 0.17 to 0.84). CONCLUSION: The concurrent use of statins during the treatment of patients with DLBCL and FL in the rituximab era did not adversely affect outcome. The apparent benefit of statin therapy on FL outcome requires further studies. PMID- 20008639 TI - Merkel cell carcinoma in the peripheral blood of a patient with concomitant chronic lymphocytic leukemia and multiple myeloma. PMID- 20008640 TI - Prognostic role of KRAS and BRAF in stage II and III resected colon cancer: results of the translational study on the PETACC-3, EORTC 40993, SAKK 60-00 trial. AB - PURPOSE: Mutations within the KRAS proto-oncogene have predictive value but are of uncertain prognostic value in the treatment of advanced colorectal cancer. We took advantage of PETACC-3, an adjuvant trial with 3,278 patients with stage II to III colon cancer, to evaluate the prognostic value of KRAS and BRAF tumor mutation status in this setting. PATIENTS AND METHODS: Formalin-fixed paraffin embedded tissue blocks (n = 1,564) were prospectively collected and DNA was extracted from tissue sections from 1,404 cases. Planned analysis of KRAS exon 2 and BRAF exon 15 mutations was performed by allele-specific real-time polymerase chain reaction. Survival analyses were based on univariate and multivariate proportional hazard regression models. RESULTS: KRAS and BRAF tumor mutation rates were 37.0% and 7.9%, respectively, and were not significantly different according to tumor stage. In a multivariate analysis containing stage, tumor site, nodal status, sex, age, grade, and microsatellite instability (MSI) status, KRAS mutation was associated with grade (P = .0016), while BRAF mutation was significantly associated with female sex (P = .017), and highly significantly associated with right-sided tumors, older age, high grade, and MSI-high tumors (all P < 10(-4)). In univariate and multivariate analysis, KRAS mutations did not have a major prognostic value regarding relapse-free survival (RFS) or overall survival (OS). BRAF mutation was not prognostic for RFS, but was for OS, particularly in patients with MSI-low (MSI-L) and stable (MSI-S) tumors (hazard ratio, 2.2; 95% CI, 1.4 to 3.4; P = .0003). CONCLUSION: In stage II-III colon cancer, the KRAS mutation status does not have major prognostic value. BRAF is prognostic for OS in MS-L/S tumors. PMID- 20008641 TI - Association between disease-free survival and overall survival when survival is prolonged after recurrence in patients receiving cytotoxic adjuvant therapy for colon cancer: simulations based on the 20,800 patient ACCENT data set. AB - PURPOSE: We previously validated disease-free survival (DFS) as a surrogate for overall survival (OS) in fluorouracil-based adjuvant colon cancer clinical trials. New therapies have extended survival after recurrence from 1 to approximately 2 years. We examined the possible impact of this improvement on the DFS/OS association. METHODS: The Adjuvant Colon Cancer Endpoints (ACCENT) data set of 20,898 patients was analyzed. In an exploratory fashion, time from recurrence to death in patients experiencing recurrence was extended using several algorithms, and the association of DFS after 3 years of median follow-up and OS after varying lengths of follow-up (median of 5, 6, and 7 years) was assessed. RESULTS: Seven thousand four hundred two patients (35%) experienced recurrence. Median time from recurrence to death was 24 months in the hypothetical data sets. When times from recurrence to death were doubled, the association between treatment effects on DFS and 5-year OS was modest (R(2) = 0.51 for both 2- and 3-year DFS) but remained strong for DFS and 6-year OS (R(2) = 0.67 for both 2- and 3-year DFS) and 7-year OS (R(2) = 0.70 for both 2- and 3 year DFS). The reduced DFS/OS association with extended survival after recurrence was greater in stage II than stage III patients. Multiple simulations provided consistent findings. CONCLUSION: Extended survival after recurrence reduces the association between treatment effects on 3-year DFS and 5-year OS, particularly in stage II patients; longer follow-up strengthens the association. In modern adjuvant trials, 6 or 7 years may be required to demonstrate OS improvements, further supporting DFS as the preferred primary end point for future adjuvant colon cancer clinical trials. PMID- 20008642 TI - Allogeneic hematopoietic stem-cell transplantation for patients 50 years or older with myelodysplastic syndromes or secondary acute myeloid leukemia. AB - PURPOSE: This study was performed to examine the characteristics of transplant activity for patients with myelodysplastic syndromes (MDS) older than 50 years within the European Group for Blood and Marrow Transplantation, and to evaluate the factors predicting outcome within this group of patients. PATIENTS AND METHODS: We performed a retrospective multicenter analysis of 1,333 MDS patients age 50 years or older who received transplantation within the EBMT since 1998. The median recipient age was 56 years, with 884 patients (66%) age 50 to 60 years and 449 (34%) patients older than 60 years. There were 811 HLA-matched sibling (61%) and 522 (39%) unrelated donor transplants. Five hundred patients (38%) received standard myeloablative conditioning (SMC), and 833 (62%) received reduced intensity conditioning (RIC). RESULTS: The 4-year estimate for overall survival of the whole cohort was 31%. On multivariate analysis, use of RIC (hazard ratio [HR], 1.44; 95% CI, 1.13 to 1.84; P < .01) and advanced disease stage at transplantation (HR, 1.51; 95% CI, 1.18 to 1.93; P < .01) were associated with an increased relapse rate. In contrast, advanced disease stage at transplantation (HR, 1.43; 95% CI, 1.13 to 1.79; P = .01), use of an unrelated donor (P = .03), and RIC (HR, 0.79; 95% CI, 0.65 to 0.97; P = .03) were independent variables associated with nonrelapse mortality. Advanced disease stage at transplantation (HR, 1.55; 95% CI, 1.32 to 1.83; P < .01) was the major independent variable associated with an inferior 4-year overall survival. CONCLUSION: Allogeneic hematopoietic stem-cell transplantation remains a potential curative therapeutic option for many older patients with MDS. In this analysis, disease stage at time of transplantation, but not recipient age or the intensity of the conditioning regimens, was the most important factor influencing outcomes. PMID- 20008643 TI - Establishing the predictive validity of intentions to smoke among preadolescents and adolescents surviving cancer. AB - PURPOSE: A significant proportion of adults surviving childhood cancer are smokers. Although these estimated rates of smoking are slightly lower than those in the US population, they remain alarmingly high for this high-risk group. The purpose of this study was to examine the predictive validity of adolescent self reported smoking intentions for later smoking among childhood cancer survivors. PATIENTS AND METHODS: Baseline tobacco intentions were collected from 119 nonsmoking cancer survivors, age 10 to 18 years, who participated in a tobacco based clinical trial during the late 1990s. Follow-up smoking status was systematically collected annually up to 10 years postintervention (median follow up, 6.0 years; interquartile range, 3.0 to 6.9 years) as part of clinical survivorship care. RESULTS: Twenty-seven participants (22.7%) subsequently initiated tobacco use within 5 years of study enrollment. The 5-year cumulative incidence was 29.8% +/- 6.0% for those who were susceptible to smoking compared with 12.8% +/- 5.4% for those who were committed never smokers (P = .022). Past use (P < .001) and having friends who smoked (P = .038) were also associated (univariate model) with tobacco initiation, and there was a trend for an association for older adolescents (P = .073). Every unit increase on the intentions scale was associated with a 17% increase in the risk for tobacco initiation (P = .002) after adjusting for age group and past tobacco use in a multivariable model. CONCLUSION: Because early intentions to smoke are predictive of later tobacco use, survivors as young as 10 years of age who waver in their commitment to remain tobacco abstinent should be targeted for tobacco prevention interventions. PMID- 20008644 TI - Efficacy and safety of pazopanib in patients with metastatic renal cell carcinoma. AB - PURPOSE: Inactivation of the von Hippel-Lindau gene in clear-cell renal cell carcinomas (RCC) leads to overexpression of hypoxia inducible factor, a transcription factor regulating vascular endothelial growth factor (VEGF) and platelet-derived growth factor (PDGF) gene expression. Pazopanib, an angiogenesis inhibitor targeting VEGF receptor, PDGF receptor, and c-KIT, was evaluated in patients with RCC. PATIENTS AND METHODS: This phase II study was designed as a randomized discontinuation study but was revised to an open-label study on the recommendation of the data monitoring committee (based on week 12 response rate [RR] of 38% in the first 60 patients). The primary end point was changed from progressive disease rate at 16 weeks postrandomization to RR. Pazopanib 800 mg was administered orally once daily. Pazopanib 800 mg was administered orally once daily. RESULTS: The study enrolled 225 patients with metastatic RCC; 155 patients (69%) were treatment naive, and 70 patients (31%) had received one prior cytokine or bevacizumab-containing regimen. Overall RR was 35%; median duration of response was 68 weeks. Median progression-free survival (PFS) was 52 weeks. Eastern Cooperative Oncology Group performance status of 0 and time from diagnosis to treatment of more than 1 year were correlated with prolonged PFS. Pazopanib was generally well tolerated. The most common adverse events were diarrhea, fatigue, and hair depigmentation. The most common laboratory abnormalities were elevated AST and ALT. CONCLUSION: Pazopanib demonstrated durable activity in patients with advanced RCC and was generally well tolerated in this population. These findings support the further development of pazopanib in advanced RCC. PMID- 20008645 TI - Pathologic complete response rates in young women with BRCA1-positive breast cancers after neoadjuvant chemotherapy. AB - PURPOSE: To estimate the rate of pathologic complete response (pCR) to neoadjuvant chemotherapy in BRCA1 mutation carriers according to chemotherapy regimen. PATIENTS AND METHODS: From a registry of 6,903 patients, we identified 102 women who carried a BRCA1 founder mutation and who had been treated for breast cancer with neoadjuvant chemotherapy. Pathologic complete response was evaluated using standard criteria. RESULTS: Twenty-four (24%) of the 102 BRCA1 mutation carriers experienced a pCR. The response rate varied widely with treatment: a pCR was observed in one (7%) of 14 women treated with cyclophosphamide, methotrexate, and fluorouracil (CMF); in two (8%) of 25 women treated with doxorubicin and docetaxel (AT); in 11 (22%) of 51 women treated with doxorubicin and cyclophosphamide (AC) or fluorouracil, doxorubicin, and cyclophosphamide (FAC), and in 10 (83%) of 12 women treated with cisplatin. CONCLUSION: A low rate of pCR was observed in women with breast cancer and a BRCA1 mutation who were treated with AT or CMF. A high rate of pCR was seen after treatment with cisplatin. An intermediate rate of PCR was associated with AC or FAC. The relative benefits of AC and platinum therapy need to be confirmed through follow-up of this and other cohorts. PMID- 20008646 TI - Effects of intensive medical therapy on microemboli and cardiovascular risk in asymptomatic carotid stenosis. AB - OBJECTIVE: To assess the effect of more intensive medical therapy on the rate of transcranial Doppler (TCD) microemboli and cardiovascular events in patients with asymptomatic carotid stenosis (ACS). DESIGN: A prospective study. SETTING: A teaching hospital. PATIENTS: Four hundred sixty-eight patients with ACS greater than 60% by Doppler peak velocity. MAIN OUTCOME MEASURES: We compared (1) the proportion of ACS patients who had microemboli on TCD, (2) cardiovascular events, (3) rate of carotid plaque progression, and (4) baseline medical therapy, before and since 2003. RESULTS: Among 468 ACS patients, 199 were enrolled between January 1, 2000, and December 31, 2002; and 269 were enrolled between January 1, 2003, and July 30, 2007. Microemboli were present in 12.6% before 2003 and 3.7% since 2003 (P < .001). The decline in microemboli coincided with better control of plasma lipids and slower progression of carotid total plaque area. Since 2003, there have been significantly fewer cardiovascular events among patients with ACS: 17.6% had stroke, death, myocardial infarction, or carotid endarterectomy for symptoms before 2003, vs 5.6% since 2003 (P < .001). The rate of carotid plaque progression in the first year of follow-up has declined from 69 mm(2) (SD, 96 mm(2)) to 23 mm(2) (SD, 86 mm(2)) (P < .001). CONCLUSIONS: Cardiovascular events and microemboli on TCD have markedly declined with more intensive medical therapy. Less than 5% of patients with ACS now stand to benefit from revascularization; patients with ACS should receive intensive medical therapy and should only be considered for revascularization if they have microemboli on TCD. PMID- 20008647 TI - Natural oxidant balance in Parkinson disease. PMID- 20008648 TI - Mild cognitive impairment: ten years later. AB - In the past 10 years, there has been a virtual explosion in the literature concerning the construct of mild cognitive impairment. The interest in this topic demonstrates the increasing emphasis on the identification of the earliest features of cognitive disorders such as Alzheimer disease and other dementias. Mild cognitive impairment represents the earliest clinical features of these conditions and, hence, has become a focus of clinical, epidemiologic, neuroimaging, biomarker, neuropathological, disease mechanism, and clinical trials research. This review summarizes the progress that has been made while also recognizing the challenges that remain. PMID- 20008649 TI - Cellular mechanisms of central nervous system repair by natural autoreactive monoclonal antibodies. AB - Natural autoreactive monoclonal IgM antibodies have demonstrated potential as therapeutic agents for central nervous system (CNS) disease. These antibodies bind surface antigens on specific CNS cells, activating intracellular repair promoting signals. IgM antibodies that bind to surface antigens on oligodendrocytes enhanced remyelination in animal models of multiple sclerosis. IgM antibodies that bind to neurons stimulate neurite outgrowth and prevent neuron apoptosis. The neuron-binding IgM antibodies may have utility in CNS axon- or neuron-damaging diseases, such as amyotrophic lateral sclerosis, stroke, spinal cord injury, or secondary progressive multiple sclerosis. Recombinant remyelination-promoting IgM antibodies have been generated for formal toxicology studies and, after Food and Drug Administration approval, a phase 1 clinical trial. Natural autoreactive monoclonal antibodies directed against CNS cells represent novel therapeutic molecules to induce repair of the nervous system. PMID- 20008650 TI - Pittsburgh compound B imaging and prediction of progression from cognitive normality to symptomatic Alzheimer disease. AB - OBJECTIVE: To determine whether preclinical Alzheimer disease (AD), as detected by the amyloid-imaging agent Pittsburgh Compound B (PiB) in cognitively normal older adults, is associated with risk of symptomatic AD. DESIGN: A longitudinal cohort study of cognitively normal older adults assessed with positron emission tomography (PET) to determine the mean cortical binding potential for PiB and followed up with annual clinical and cognitive assessments for progression to very mild dementia of the Alzheimer type (DAT). SETTING: The Alzheimer's Disease Research Center, Washington University, St Louis, Missouri. PARTICIPANTS: One hundred fifty-nine participants with a mean age of 71.5 years with a Clinical Dementia Rating (CDR) of 0 on a PET PiB scan at baseline. MAIN OUTCOME MEASURE: Progression from CDR 0 to CDR 0.5 status (very mild dementia). RESULTS: Twenty three participants progressed to CDR 0.5 at follow-up assessment (range, 1-5 assessments after PET PiB). Of these, 9 also were diagnosed with DAT. Higher mean cortical binding potential values for PiB (hazard ratio, 4.85; 95% confidence interval, 1.22-19.01; P = .02) and age (hazard ratio, 1.14; 95% confidence interval, 1.02-1.28; P = .03) predicted progression to CDR 0.5 DAT. The CDR 0.5 DAT group showed decline in 3 cognitive domains (episodic memory, semantic memory, and visuospatial performance) and had volume loss in the parahippocampal gyrus (includes entorhinal cortex) compared with individuals who remained at CDR 0. CONCLUSION: Preclinical AD as detected by PET PiB is not benign, as it is associated with progression to symptomatic AD. PMID- 20008651 TI - Cognitive decline and brain volume loss as signatures of cerebral amyloid-beta peptide deposition identified with Pittsburgh compound B: cognitive decline associated with Abeta deposition. AB - OBJECTIVE: To examine the relation of amyloid-beta peptide (Abeta) levels in the cerebral cortex with structural brain integrity and cognitive performance in cognitively healthy older people. DESIGN: Longitudinal study from May 22, 1985, through October 15, 2008. SETTING: Washington University Alzheimer Disease Research Center. PARTICIPANTS: A total of 135 individuals aged 65 to 88 years with a Clinical Dementia Rating of 0. MAIN OUTCOME MEASURES: The relations between mean cortical carbon 11 ((11)C)-labeled Pittsburgh compound B (PiB) binding potential values, proportional to the density of fibrillar Abeta binding sites in the brain, concurrent regional brain volumes as assessed by magnetic resonance imaging, and both concurrent and longitudinal cognitive performance in multiple domains. RESULTS: Elevated cerebral Abeta levels, in some cases comparable to those seen in individuals with Alzheimer disease, were observed in 29 participants, who also had smaller regional volumes in the hippocampus, temporal neocortex, anterior cingulate, and posterior cingulate. Concurrent cognitive performance was unrelated to Abeta levels but was related to regional brain volumes with the exception of the caudate. Longitudinal cognitive decline in episodic and working memory and visuospatial ability was associated with elevated Abeta levels and decreased hippocampal volume. CONCLUSION: The in vivo measure of cerebral amyloidosis known as [(11)C]PiB is associated with cross sectional regionally specific brain atrophy and longitudinal cognitive decline in multiple cognitive domains that occur before the clinical diagnosis of Alzheimer disease. These findings contribute to the understanding of the cognitive and structural consequences of Abeta levels in cognitively healthy older adults. PMID- 20008652 TI - Transactive response DNA-binding protein 43 burden in familial Alzheimer disease and Down syndrome. AB - OBJECTIVE: To assess the transactive response DNA-binding protein 43 (TDP-43) burden in familial forms of Alzheimer disease (FAD) and Down syndrome (DS) to determine whether TDP-43 inclusions are also present. DESIGN: Using standard immunohistochemical techniques, we examined brain tissue samples from 42 subjects with FAD and 14 with DS. RESULTS: We found pathological TDP-43 aggregates in 14.0% of participants (6 of 42 and 2 of 14 participants with FAD and DS, respectively). In both FAD and DS, TDP-43 immunoreactivity did not colocalize with neurofibrillary tangles. Occasionally participants with FAD or DS had TDP-43 positive neuropil threads or dots. Overall, the amygdala was most commonly affected, followed by the hippocampus, with no TDP-43 pathology in neocortical regions. A similar distribution of TDP-43 inclusions is seen in sporadic Alzheimer disease, but it differs from that seen in amyotrophic lateral sclerosis and frontotemporal dementia. CONCLUSIONS: Transactive response DNA-binding protein 43 pathology occurs in FAD and DS, similar to that observed in sporadic Alzheimer disease. Thus, pathological TDP-43 may contribute the cognitive impairments in familial and sporadic forms of Alzheimer disease. PMID- 20008653 TI - Characteristics and surgical outcomes of patients with refractory magnetic resonance imaging-negative epilepsies. AB - OBJECTIVE: To explore several characteristics of patients with pharmacoresistant epilepsy without distinct lesions on magnetic resonance images (MRI(-)), who account for a relevant proportion of presurgical patient cohorts. DESIGN: Retrospective case series. SETTING: University epilepsy center. PATIENTS: A cohort of 1200 patients who had comprehensive presurgical assessment from January 1, 2000, through December 31, 2006. MAIN OUTCOME MEASURES: Frequency of MRI(-) patients in the total presurgical cohort, seizure-free outcome rates in patients who had surgery and those who did not, outcome predictors, and spatial properties of epileptogenic areas in MRI(-) patients with epilepsy. All MRI(-) patients were retrospectively analyzed. Presurgical MRIs were reevaluated for subtle cortical dysplasias by postprocessing and visual reassessment. RESULTS: One-hundred ninety MRI(-) patients were identified (16% of all presurgical candidates); 29 (15%) had surgery. Eleven (38%) became seizure free (including those with auras only; 45%). Surgical therapy was more frequently offered to MRI(+) patients (76%; P < .001), and their outcome was also superior (66% seizure-free; P = .001). The seizure free rate of 16% in MRI(-) patients who did not have surgery was, however, inferior to that of the MRI(-) patients who did (P = .008). Nine MRI(-) patients who had surgery had distinct histopathological lesions, 8 of which turned out to be retrospectively detectable on presurgical MRI. Seven of the MRI(-) but histopathologically lesional patients became seizure free compared with only 4 of 20 patients without histopathological lesions (P = .003). Three-fifths of the histopathologically nonlesional patients had multifocal or extensive epileptogenic areas. CONCLUSIONS: Patients with epilepsy who are MRI(-) can be successfully treated with surgery. Improved sensitivity of MRI will improve the outcomes of presurgically studied patients. Surgical failures in patients without histopathological lesions mostly result from extensive epileptogenic areas. PMID- 20008654 TI - Status epilepticus associated with subtentorial posterior fossa lesions. AB - BACKGROUND: Nonconvulsive status epilepticus (SE) is a frequent complication in critically ill patients in the intensive care unit. While seizures have been reported in association with subtentorial posterior fossa lesions, the frequency of occurrence of SE among these patients is not known. OBJECTIVES: To examine prevalence, clinical features, potential risk factors, and outcome of SE among patients presenting with subtentorial posterior fossa lesions. DESIGN: Retrospective review of our hospital database was conducted to identify patients with posterior fossa lesions complicated by SE over 1 year between April 1, 2007, and May 1, 2008. SETTING: Tertiary care setting. PATIENTS: Patients with subtentorial posterior fossa lesions admitted to the hospital for neurological or neurosurgical care. MAIN OUTCOME MEASURES: Prevalence of SE, potential risk factors, and eventual neurological outcome. RESULTS: Over 1 year, 13 of 501 patients (2.6%) admitted to the hospital with posterior fossa lesions had SE. Some patients had risk factors for SE such as sepsis, use of particular drugs, or intracranial bleeding, while others had no other clear identifiable cause. CONCLUSIONS: Status epilepticus can be a potential complication in patients with posterior fossa cranial lesions and can be seen in up to 2.6% of such patients. Most have unfavorable outcome. PMID- 20008655 TI - Functional and cognitive outcome in prolonged refractory status epilepticus. AB - OBJECTIVE: To determine the functional and cognitive outcomes of patients with prolonged refractory status epilepticus (PRSE) lasting 7 or more days despite the use of anesthetic agents for seizure suppression. DESIGN: Retrospective analysis. SETTING: St Mary's Hospital, Mayo Clinic, Rochester, Minnesota. PARTICIPANTS: Fourteen patients with PRSE. INTERVENTION: Hospital follow-up interview. MAIN OUTCOME MEASURES: Survival rate of PRSE and functional and cognitive outcome of surviving patients based on the modified Rankin Scale (mRS) and Telephone Interview for Cognitive Status (TICS). RESULTS: Forty-three percent of patients (6 of 14) died during hospitalization for PRSE, and 57% (8 of 14) had died by the last follow-up. Of the 6 surviving patients, 4 showed improvement and 2 showed no change in mRS score (median mRS change, -1; range, 0 to -3). Owing to preexisting cognitive deficits, 1 patient could not complete the TICS. The 5 remaining patients scored a median of 34 on the TICS (range, 30-37; reference TICS score, >or=31; maximum TICS score, 41). Age, sex, PRSE duration, and etiology were not associated with chance of survival. CONCLUSIONS: Despite the high mortality rate, survival with meaningful functional and cognitive recovery is possible after PRSE. Prolonged duration of status epilepticus alone should not be considered a reason to discontinue treatment. PMID- 20008656 TI - Genotype-phenotype correlations in Charcot-Marie-Tooth disease type 2 caused by mitofusin 2 mutations. AB - BACKGROUND: Mutations in the gene encoding mitofusin 2 (MFN2) cause Charcot-Marie Tooth disease type 2 (CMT2), with heterogeneity concerning severity and associated clinical features. OBJECTIVE: To describe MFN2 mutations and associated phenotypes in patients with hereditary motor and sensory neuropathy (HMSN). DESIGN: Direct sequencing of the MFN2 gene and clinical investigations of patients with MFN2 mutations. SETTING: Molecular genetics laboratory of a university hospital and the Limoges National Referral Center for Rare Peripheral Neuropathies. PATIENTS: One hundred fifty index patients with HMSN and a median motor nerve conduction velocity of 25 m/s or greater and without mutations in the genes encoding connexin 32 and myelin protein zero. MAIN OUTCOME MEASURES: Results of genetic analyses and phenotypic observations. RESULTS: Twenty different missense mutations were identified in 20 index patients. Mutation frequency was 19 of 107 (17.8%) in patients with CMT2 and 1 of 43 (2.3%) in patients with a median motor nerve conduction velocity less than 38 m/s. Four patients had proven de novo mutations, 8 families had autosomal dominant inheritance, and 3 had autosomal recessive inheritance. The remaining 5 patients were sporadic cases with heterozygous mutations. Phenotypes varied from mild forms to early-onset severe forms. Additional features were encountered in 8 patients (32%). Six patients underwent sural nerve biopsy: electronic microscopy showed prominent mitochondrial abnormalities on longitudinal sections. CONCLUSIONS: MFN2 mutations are a frequent cause of CMT2, with variable severity and either dominant or recessive inheritance. MFN2 gene testing must be a first line analysis in axonal HMSN irrespective of the mode of inheritance or the severity of the peripheral neuropathy. PMID- 20008658 TI - Heterogeneity in infarct patterns and clinical outcomes following internal carotid artery occlusion. AB - OBJECTIVE: To investigate whether the extent of infarction and clinical outcomes after internal carotid artery (ICA) occlusion depends on the additional occlusion of the middle cerebral artery (MCA). DESIGN: Using statistical parametric mapping, we compared infarct patterns in stroke patients. SETTING: A tertiary care hospital. PATIENTS: Patients with coexistent ICA and MCA occlusion (n = 25), isolated ICA occlusion (n = 20), and isolated MCA occlusion (n = 40). MAIN OUTCOME MEASURE: Modified Rankin scale score. The independent effect of infarct type on clinical outcome was estimated using logistic regression, adjusting for age and sex. RESULTS: The mean age was 62.6 years (standard deviation [SD], 15.5 years) in patients with ICA and MCA occlusion, 64.3 years (SD, 12.9 years) in patients with isolated ICA occlusion, and 67.4 years (SD, 14.2 years) in patients with isolated MCA occlusion. Infarct patterns, volume (P = .13), and the proportion of patients with poor outcomes (P = .5) were similar between those with ICA and MCA occlusions and those with isolated MCA occlusion. Compared with the other 2 groups, those with isolated ICA occlusion were less likely to have infarction of the insula (P < .001) and superior temporal lobe (P < .001) and had smaller infarct volume and lower modified Rankin scale scores (all P < .05). Compared with those with isolated ICA occlusion, the risk of poor clinical outcome was greater in those with coexistent ICA and MCA occlusion (P = .02) and those with isolated MCA occlusion (P = .06) independent of age and sex. COMMENTS: Patients with ICA occlusion but without coexistent MCA occlusion have different infarct patterns, less extensive infarcts, and better clinical outcomes than those with coexistent MCA occlusion or MCA occlusion alone. It may not be warranted to exclude such patients from acute stroke trials. PMID- 20008657 TI - Motor phenotype of LRRK2 G2019S carriers in early-onset Parkinson disease. AB - OBJECTIVE: To determine the motor phenotype of LRRK2 G2019S mutation carriers. LRRK2 mutation carriers were previously reported to manifest the tremor dominant motor phenotype, which has been associated with slower motor progression and less cognitive impairment compared with the postural instability and gait difficulty (PIGD) phenotype. DESIGN: Cross-sectional observational study. SETTING: Thirteen movement disorders centers. PARTICIPANTS: Nine hundred twenty-five early-onset Parkinson disease cases defined as age at onset younger than 51 years. MAIN OUTCOME MEASURES: LRRK2 mutation status and Parkinson disease motor phenotype: tremor dominant or PIGD. Demographic information, family history of Parkinson disease, and the Unified Parkinson's Disease Rating Scale score were collected on all participants. DNA samples were genotyped for LRRK2 mutations (G2019S, I2020T, R1441C, and Y1699C). Logistic regression was used to examine associations of G2019S mutation status with motor phenotype adjusting for disease duration, Ashkenazi Jewish ancestry, levodopa dose, and family history of Parkinson disease. RESULTS: Thirty-four cases (3.7%) (14 previously reported) were G2019S carriers. No other mutations were found. Carriers were more likely to be Ashkenazi Jewish (55.9% vs 11.9%; P < .001) but did not significantly differ in any other demographic or disease characteristics. Carriers had a lower tremor score (P = .03) and were more likely to have a PIGD phenotype (92.3% vs 58.9%; P = .003). The association of the G2019S mutation with PIGD phenotype remained after controlling for disease duration and Ashkenazi Jewish ancestry (odds ratio, 17.7; P < .001). CONCLUSION: Early-onset Parkinson disease G2019S LRRK2 carriers are more likely to manifest the PIGD phenotype, which may have implications for disease course. PMID- 20008659 TI - HLA-DRB1*1501 and spinal cord magnetic resonance imaging lesions in multiple sclerosis. AB - BACKGROUND: Multiple sclerosis (MS) is a heterogeneous neurologic disease with extensive variation with respect to the most affected central nervous system region (brain vs spinal cord). OBJECTIVE: To test the hypothesis that this variation in lesion location (brain vs spinal cord) might be (partially) genetically determined. DESIGN: Candidate gene study. SETTING: Academic research. PATIENTS: Patients were selected for the availability of DNA material, clinical variables, and brain and spinal cord magnetic resonance images (evaluating T2 weighted lesion load in the brain and the number of spinal cord lesions). MAIN OUTCOME MEASURES: For genotyping, we used a DNA chip containing a set of genes mentioned in previous publications noting their relation to different phenotypes of MS. We assessed the association between brain and spinal cord abnormalities and the genotypes of the patients. RESULTS: One hundred fifty patients were included in the analysis. Five single-nucleotide polymorphisms within the major histocompatibility complex region were associated with the number of focal abnormalities in the spinal cord. The most significant was rs3135388 (surrogate marker for the HLA-DRB1*1501 allele). Carriers of HLA-DRB1*1501 had a median of 4 spinal cord lesions compared with 2 lesions for noncarriers (P < .001). No significant association was noted between the single-nucleotide polymorphisms and T2-weighted lesion load in the brain. CONCLUSIONS: Carriership of HLA-DRB1*1501 (via rs3135388) was associated with the extent of focal abnormalities in the spinal cord. Spinal cord lesions might be an explanation for increased MS disease severity in patients carrying HLA-DRB1*1501. PMID- 20008660 TI - High striatal amyloid beta-peptide deposition across different autosomal Alzheimer disease mutation types. AB - BACKGROUND: Supported by compelling genetic data regarding early-onset familial Alzheimer disease (AD), the amyloid beta-peptide (Abeta)-centric theory holds that Abeta is involved in the pathogenesis of sporadic AD. Mutations in the amyloid precursor protein (APP), presenilin 1 (PSEN1), and presenilin 2 (PSEN2) genes lead to increased Abeta levels before symptoms arise. OBJECTIVES: To evaluate the pattern of Pittsburgh Compound B (PiB) retention in subjects with different autosomal dominant mutations associated with familial AD vs that in healthy age-matched control subjects and subjects with probable sporadic AD, to correlate Abeta burden as measured by PiB with available clinical and cognitive data, and to compare the regional brain patterns of PiB retention and fluorodeoxyglucose F 18 (FDG) uptake. DESIGN: Correlation analysis of positron emission tomography (PET) imaging studies. SETTING: Academic research. PARTICIPANTS: Seven PSEN1 mutation carriers and 1 APP mutation carrier underwent PiB and FDG PET imaging. Amyloid beta-peptide burden and FDG uptake were established using standardized uptake values normalized to pons. MAIN OUTCOME MEASURE: Primary outcomes were PET results, which were compared with those of a well-characterized cohort of 30 healthy control subjects and 30 subjects with probable sporadic AD. RESULTS: All mutation carriers had high PiB retention in the striatum, with some also having cortical PiB retention in ventrofrontal and posterior cingulate/precuneus areas. The striatal pattern of PiB retention was similar in the PSEN1 and APP mutation carriers. Neither striatal nor cortical Abeta burden was related to cognitive status. CONCLUSIONS: Consistent with previous studies, the pattern of Abeta deposition in familial AD differs from that in sporadic AD, with higher striatal and somewhat lower cortical PiB retention in familial AD. The pattern and degree of Abeta deposition were not associated with mutation type nor cognitive status. PMID- 20008661 TI - Quantitative template for subtyping primary progressive aphasia. AB - BACKGROUND: The syndrome of primary progressive aphasia (PPA) is diagnosed when a gradual failure of word usage or comprehension emerges as the principal feature of a neurodegenerative disease. OBJECTIVE: To provide a quantitative algorithm for classifying PPA into agrammatic (PPA-G), semantic (PPA-S), and logopenic (PPA L) variants, each of which is known to have a different probability of association with Alzheimer disease vs frontotemporal lobar degeneration. DESIGN: Prospective study. SETTING: University medical center. PATIENTS: Sixteen consecutively enrolled patients with PPA who underwent neuropsychological testing and magnetic resonance imaging recruited nationally in the United States as part of a longitudinal study. RESULTS: A 2-dimensional template that reflects performance on tests of syntax (Northwestern Anagram Test) and lexical semantics (Peabody Picture Vocabulary Test-Fourth Edition) classified all 16 patients in concordance with a clinical diagnosis that had been made before the administration of quantitative tests. All 3 PPA subtypes had distinctly asymmetrical atrophy of the left perisylvian language network. Each subtype also had distinctive peak atrophy sites: PPA-G in the inferior frontal gyrus (Broca area), PPA-S in the anterior temporal lobe, and PPA-L in Brodmann area 37. CONCLUSIONS: Once an accurate root diagnosis of PPA is made, subtyping can be quantitatively guided using a 2-dimensional template based on orthogonal tasks of grammatical competence and word comprehension. Although the choice of tasks and the precise cutoff levels may need to be adjusted to fit linguistic and educational backgrounds, these 16 patients demonstrate the feasibility of using a simple algorithm for clinicoanatomical classification in PPA. Prospective studies will show whether this subtyping can improve clinical prediction of the underlying neuropathologic condition. PMID- 20008662 TI - Cerebrospinal fluid beta-amyloid 42, Tau, and P-tau: confirmation now realization. PMID- 20008663 TI - Surgery is the best option for intractable unilateral mesial temporal epilepsy. PMID- 20008665 TI - Symptomatic narcolepsy in patients with neuromyelitis optica and multiple sclerosis: new neurochemical and immunological implications. AB - OBJECTIVE: To characterize factors that contribute to symptomatic narcolepsy and excessive daytime sleepiness in neuromyelitis optica and multiple sclerosis. SETTING: Japanese university hospitals. DESIGN: Case study. PATIENTS: Seven Japanese patients whose initial diagnoses were multiple sclerosis and who were exhibiting excessive daytime sleepiness. MAIN OUTCOME MEASURES: Lesions on magnetic resonance imaging, cerebrospinal fluid hypocretin-1 levels, and serum anti-aquaporin 4 (AQP4) antibody titer. RESULTS: Bilateral and symmetrical hypothalamic lesions associated with marked or moderate hypocretin deficiency were found in all 7 cases. Four of these patients met the International Classification of Sleep Disorders 2 narcolepsy criteria. Three patients, including 2 patients with narcolepsy, were seropositive for anti-AQP4 antibody and diagnosed as having neuromyelitis optica-related disorder. CONCLUSION: Since AQP4 is highly expressed in the hypothalamic periventricular regions, an immune attack on AQP4 may be partially responsible for the bilateral and hypothalamic lesions and hypocretin deficiency in narcolepsy/excessive daytime sleepiness associated with autoimmune demyelinating diseases. PMID- 20008664 TI - Absence of Pittsburgh compound B detection of cerebral amyloid beta in a patient with clinical, cognitive, and cerebrospinal fluid markers of Alzheimer disease: a case report. AB - BACKGROUND: To date, there have been no reports of individuals who have been characterized longitudinally using clinical and cognitive measures and who transitioned from cognitive normality to early symptomatic Alzheimer disease (AD) during a period when both cerebrospinal fluid (CSF) markers and Pittsburgh Compound B (PiB) amyloid imaging were obtained. OBJECTIVE: To determine the temporal relationships of clinical, cognitive, CSF, and PiB amyloid imaging markers of AD. DESIGN: Case report. SETTING: Alzheimer disease research center. PARTICIPANT: Longitudinally assessed 85-year-old man in a memory and aging study who was cognitively normal at his initial and next 3 annual assessments. MAIN OUTCOME MEASURES: Serial clinical and psychometric assessments over 6 years in addition to PiB imaging with positron emission tomography (PET) and CSF biomarker assays before autopsy. RESULTS: Decline in measures of episodic memory and, to a lesser degree, working memory began at about age 88 years. PiB PET amyloid imaging was negative at age 88(1/2) years, but at age 89(1/2) years there was reduced amyloid beta 42 and elevated levels of tau in the CSF. Beginning at age 89 years, very mild cognitive and functional decline reported by his collateral source resulted in a diagnosis of very mild dementia of the Alzheimer type. After death at age 91 years, the autopsy revealed foci of frequent neocortical diffuse amyloid beta plaques sufficient to fulfill Khachaturian neuropathologic criteria for definite AD, but other neuropathologic criteria for AD were not met because only sparse neuritic plaques and neurofibrillary tangles were present. Postmortem biochemical analysis of the cerebral tissue confirmed that PiB PET binding was below the level needed for in vivo detection. CONCLUSION: Clinical, cognitive, and CSF markers consistent with AD may precede detection of cerebral amyloid beta using amyloid imaging agents such as PiB that primarily label fibrillar amyloid beta plaques. PMID- 20008666 TI - Subdural fluid collections in patients with infantile neuronal ceroid lipofuscinosis. AB - OBJECTIVE: To describe subdural fluid collections on magnetic resonance imaging as part of the natural history of infantile neuronal ceroid lipofuscinosis. DESIGN: Case series. SETTING: Program on Developmental Endocrinology and Genetics, The Clinical Center, National Institutes of Health, Bethesda, Maryland. PATIENTS: Patients with infantile neuronal ceroid lipofuscinosis with subdural fluid collections. MAIN OUTCOME MEASURE: Neurodegeneration on magnetic resonance imaging. RESULTS: During an ongoing bench-to-bedside clinical investigation, magnetic resonance imaging examinations led to the incidental discovery of subdural fluid collections in 4 of 9 patients with infantile neuronal ceroid lipofuscinosis. No particular event (such as trauma) or change in symptoms was linked to this finding, which was already in the chronic phase when discovered. Of the 4 patients, 1 was followed up for 7 years, 2 for 4 years, and 1 for 2.5 years. Over time, these collections remained stable or decreased in size. CONCLUSION: Recognition that subdural fluid collections are part of the infantile neuronal ceroid lipofuscinosis disease process may obviate the necessity of additional workup as well as therapeutic interventions in these chronically sick children. PMID- 20008667 TI - Shivering in coma. PMID- 20008668 TI - Lateral medullary syndrome and ipsilateral hemiplegia (Opalski syndrome) due to left vertebral artery dissection. PMID- 20008669 TI - Progressive myopathy with multiple symmetric lipomatosis. PMID- 20008670 TI - Protective effect of breastfeeding in postpartum relapse rate of mothers with multiple sclerosis. PMID- 20008671 TI - Breastfeeding and multiple sclerosis. PMID- 20008672 TI - Oral carnitine therapy and insulin resistance. PMID- 20008673 TI - Cardioprotective actions of cyclic GMP: lessons from genetic animal models. PMID- 20008674 TI - Angiotensin II, oxidant signaling, and hypertension: down to a T? PMID- 20008676 TI - Is longitudinal pulse pressure a better predictor of 24-hour urinary albumin excretion than other indices of blood pressure? AB - The strong relationship between urinary albumin excretion (UAE) and pulse pressure (PP) in cross-sectional studies suggests that pressure pulsatility may contribute to renal microvascular injury. The longitudinal relationships between UAE and the various indices of blood pressure (BP) are not well studied. We compared the associations of UAE with the longitudinal exposure to PP and systolic, diastolic, and mean BPs. UAE was measured from 24-hour urine collections in 450 community-dwelling subjects (age: 57+/-15 years, 53% women, all with UAE <200 microg/min). For each subject, longitudinal indices of BP were estimated by dividing the area under the curve of serial measurements of BP (median: 5) during 1 to 22 years preceding UAE measurement by the number of follow-up years. Median (interquartile range) UAE was 4.7 microg/min (3.3 to 7.8 microg/min) in women and 5.2 microg/min (3.7 to 9.8 microg/min) in men. In women, UAE was not related to longitudinal indices of BP. In men, in multivariable adjusted models that included either longitudinal systolic and diastolic BPs or longitudinal PP and mean BP, UAE was independently associated with systolic (standardized regression coefficient [beta]=0.227; P=0.03) but not with diastolic (beta=-0.049; P=0.59) BP and with PP (beta=0.216; P=0.01) but not with mean BP (beta=0.032; P=0.72). Comparisons of these 2 models and stepwise regression analyses both indicated that, of the 4 longitudinal indices of BP, PP was the strongest predictor of UAE in men. The pulsatile component of BP confers the highest risk for BP-induced renal microvascular injury. Future studies should examine whether PP reduction provides additional renoprotection beyond that attained by conventional BP goals alone. PMID- 20008675 TI - Induction of hypertension and peripheral inflammation by reduction of extracellular superoxide dismutase in the central nervous system. AB - The circumventricular organs (CVOs) lack a well-formed blood-brain barrier and produce superoxide in response to angiotensin II and other hypertensive stimuli. This increase in central superoxide has been implicated in the regulation of blood pressure. The extracellular superoxide dismutase (SOD3) is highly expressed in cells associated with CVOs and particularly with tanycytes lining this region. To understand the role of SOD3 in the CVOs in blood pressure regulation, we performed intracerebroventricular injection an adenovirus encoding Cre recombinase (5x10(8) particles per milliliter) in mice with loxP sites flanking the SOD3 coding region (SOD3(loxp/loxp) mice). An adenovirus encoding red fluorescent protein was injected as a control. Deletion of CVO SOD3 increased baseline blood pressure modestly and markedly augmented the hypertensive response to low-dose angiotensin II (140 ng/kg per day), whereas intracerebroventricular injection of adenovirus encoding red-fluorescent protein had minimal effects on these parameters. Adenovirus encoding Cre-recombinase-treated mice exhibited increased sympathetic modulation of heart rate and blood pressure variability, increased vascular superoxide production, and T-cell activation as characterized by increased circulating CD69(+)/CD3(+) cells. Deletion of CVO SOD3 also markedly increased vascular T-cell and leukocyte infiltration caused by angiotensin II. We conclude that SOD3 in the CVO plays a critical role in the regulation of blood pressure, and its loss promotes T-cell activation and vascular inflammation, in part by modulating sympathetic outflow. These findings provide insight into how central signals produce vascular inflammation in response to hypertensive stimuli, such as angiotensin II. PMID- 20008677 TI - DC-SCRIPT: nuclear receptor modulation and prognostic significance in primary breast cancer. AB - BACKGROUND: Nuclear receptors, including estrogen receptor (ER), progesterone receptor (PR)-B, peroxisome proliferator-activated receptor gamma, and retinoic acid receptor alpha, have been implicated in breast cancer etiology and progression. We investigated the role of dendritic cell-specific transcript (DC SCRIPT) as coregulator of these nuclear receptors and as a prognostic factor in breast cancer. METHODS: The effect of DC-SCRIPT on the transcriptional activity of nuclear receptors was assessed by luciferase reporter assays. DC-SCRIPT expression in normal and tumor tissue from breast cancer patients was analyzed by polymerase chain reaction and immunohistochemistry. The prognostic value of tumor DC-SCRIPT mRNA expression was assessed in three independent cohorts of breast cancer patients: a discovery group (n = 47) and a validation group (n = 97) (neither of which had received systemic adjuvant therapy) and in a tamoxifen treated validation group (n = 68) by using a DC-SCRIPT to porphobilinogen deaminase transcript ratio cutoff of 0.15 determined in the discovery group. Univariate and multivariable Cox proportional hazards model analyses were performed. All statistical tests were two-sided. RESULTS: DC-SCRIPT suppressed ER and PR-mediated transcription in a ligand-dependent fashion, whereas it enhanced the retinoic acid receptor alpha- and peroxisome proliferator-activated receptor gamma-mediated transcription. In breast tissue samples from nine patients, DC SCRIPT mRNA was expressed at lower levels in the tumor than in the corresponding normal tissue (P = .010). Patients in the discovery group with high tumor DC SCRIPT mRNA levels (66%) had a longer disease-free interval than those with a low DC-SCRIPT mRNA level (34%) (hazard ratio [HR] of recurrence for high vs low DC SCRIPT level = 0.23, 95% confidence interval [CI] = 0.06 to 0.93, P = .039), which was confirmed in the validation group (HR of recurrence = 0.50, 95% CI = 0.26 to 0.95, P = .034). This prognostic value was confined to patients with ER- and/or PR-positive tumors (discovery group: HR of recurrence = 0.16, 95% CI = 0.03 to 0.89, P = .030; validation group: HR of recurrence = 0.42, 95% CI = 0.19 to 0.91, P = .028) and was also observed in the second validation group (HR = 0.46, 95% CI = 0.22 to 0.97, P = .040). DC-SCRIPT was an independent prognostic factor after correction for tumor size, lymph node status, and adjuvant therapy (n = 145; HR = 0.50, 95% CI = 0.29 to 0.85, P = .010). CONCLUSION: DC-SCRIPT is a key regulator of nuclear receptor activity that has prognostic value in breast cancer. PMID- 20008678 TI - Tongue pressure modulation during swallowing: water versus nectar-thick liquids. AB - PURPOSE: Evidence of tongue-palate pressure modulation during swallowing between thin and nectar-thick liquids stimuli has been equivocal. This mirrors a lack of clear evidence in the literature of tongue and hyoid movement modulation between nectar-thick and thin liquid swallows. In the current investigation, the authors sought to confirm whether tongue-palate pressures are modulated between discrete swallows of water and nectar-thick juice. METHOD: Tongue-palate pressures were measured at 3 sites (anterior, medial, and posterior palate) using an adhered 3 bulb pressure strip in 20 healthy, young adults during discrete swallows of water and nectar-thick apple juice. RESULTS: Pressure modulation was not noted with respect to pressure amplitudes (in mm Hg), but was identified both in the pressure patterns observed (the sites and number of bulbs activated) and temporal aspects of pressure duration. CONCLUSION: Tongue-palate pressure amplitude modulation does not occur for nectar-thick swallows compared to thin liquid swallows. Modulation does, however, occur with respect to the tongue-palate contact surface area and pressure durations. The authors introduce the concept of pressure slope as a meaningful way to examine tongue-palate pressure application in swallowing. PMID- 20008679 TI - The development and utilization of a scale to measure adolescents' attitudes toward peers who use augmentative and alternative communication (AAC) devices. AB - PURPOSE: Study 1 developed the Assessment of Attitudes Toward Augmentative and Alternative Communication-2 (AATAAC-2) to assess adolescents' attitudes toward peers who use AAC. Study 2 used the AATAAC-2 to examine influences of familiarity with people with disabilities; type of AAC device; and various combinations of gender of rater, AAC user, and communication partner on adolescents' attitudes. METHOD: In Study 1, 194 adolescents viewed videotapes depicting adolescents using AAC, then completed AATAAC-2. Study 2 utilized 8 videotapes depicting 4 different gender combinations of AAC user and communication partner as experimental stimuli. Each gender combination was filmed twice: once with a static touch screen device, and once with a dynamic touch screen device. One-hundred thirty six adolescents were randomly assigned to view 1 of the 8 videos. Participants then completed AATAAC-2. RESULTS: Study 1 demonstrated that AATAAC-2 has adequate psychometric properties. Raters' responses in Study 2 indicated no main effect of device type; girls were more positive than boys; and familiarity with peers with disabilities was associated with more positive attitudes. No 2-way interactions were significant; 3-way interaction of level of familiarity, gender, and type of device used was significant. CONCLUSIONS: Familiarity and gender contribute to adolescents' attitudes; type of AAC device combined with these factors to influence attitudes. PMID- 20008680 TI - Effect of dystrophin deficiency on selected intrinsic laryngeal muscles of the mdx mouse. AB - BACKGROUND: Intrinsic laryngeal muscles (ILM) show biological differences from the broader class of skeletal muscles. Yet most research regarding ILM specialization has been completed on a few muscles, most notably the thyroarytenoid and posterior cricoarytenoid. Little information exists regarding the biology of other ILM. Early evidence suggests that the interarytenoid (IA) and cricothyroid (CT) may be more similar to classic skeletal muscle than their associated laryngeal muscles. Knowledge of the IA and CT's similarity or dissimilarity to typical skeletal muscle may hold implications for the treatment of dysphonia. PURPOSE: The purpose of this study was to further define IA and CT biology by examining their response to the biological challenge of dystrophin deficiency. METHOD: Control and dystrophin-deficient superior cricoarytenoid (SCA; mouse counterpart of IA) and CT muscles were examined for fiber morphology, sarcolemmal integrity, and immunohistochemical detection of dystrophin. RESULTS: Despite the absence of dystrophin, experimental muscles did not show disease markers. CONCLUSIONS: The SCA and the CT appear spared in dystrophin-deficient mouse models. These laryngeal muscles possess specializations that separate them from typical skeletal muscle. Considered in light of previous research, the CT and IA may represent transitional form of muscle, evidencing properties of typical and specialized skeletal muscle. PMID- 20008681 TI - Influence of sound immersion and communicative interaction on the Lombard effect. AB - PURPOSE: To examine the influence of sound immersion techniques and speech production tasks on speech adaptation in noise. METHOD: In Experiment 1, we compared the modification of speakers' perception and speech production in noise when noise is played into headphones (with and without additional self-monitoring feedback) or over loudspeakers. We also examined how this sound immersion effect depends on noise type (broadband or cocktail party) and level (from 62 to 86dB SPL). In Experiment 2, we compared the modification of acoustic and lip articulatory parameters in noise when speakers interact or not with a speech partner. RESULTS: Speech modifications in noise were greater when cocktail party noise was played in headphones than over loudspeakers. Such an effect was less noticeable in broadband noise. Adding a self-monitoring feedback into headphones reduced this effect but did not completely compensate for it. Speech modifications in noise were greater in interactive situation and concerned parameters that may not be related to voice intensity. CONCLUSIONS: The results support the idea that the Lombard effect is both a communicative adaptation and an automatic regulation of vocal intensity. The influence of auditory and communicative factors has some methodological implications on the choice of appropriate paradigms to study the Lombard effect. PMID- 20008682 TI - Phonological awareness, vocabulary, and reading in deaf children with cochlear implants. AB - PURPOSE: To explore the phonological awareness skills of deaf children with cochlear implants (CIs) and relationships with vocabulary and reading development. METHOD: Forty-three deaf children with implants who were between 5 and 15 years of age were tested; 21 had been implanted at around 2.5 years of age (Early CI group), and 22 had been implanted at around 5 years of age (Late CI group). Two control groups-a deaf hearing aided group (16 children) and a typically developing group of hearing children (19 children)-were also tested. All children received a battery of phonological processing tasks along with measures of reading, vocabulary, and speechreading. Analyses focus on deaf children within the normal IQ range (n = 53). RESULTS: Age at cochlear implantation had a significant effect on vocabulary and reading outcomes when quotient scores were calculated. Individual differences in age at implant, duration of fit, phonological development, vocabulary development, auditory memory, visual memory, and speech intelligibility were all strongly associated with progress in reading for the deaf implanted children. Patterns differed somewhat depending on whether quotient scores or standard scores were used. CONCLUSIONS: Cochlear implantation is associated with development of the oral language, auditory memory, and phonological awareness skills necessary for developing efficient word recognition skills. There is a benefit of earlier implantation. PMID- 20008683 TI - Vowel acoustics in adults with apraxia of speech. AB - PURPOSE: To investigate the hypothesis that vowel production is more variable in adults with acquired apraxia of speech (AOS) relative to healthy individuals with unimpaired speech. Vowel formant frequency measures were selected as the specific target of focus. METHOD: Seven adults with AOS and aphasia produced 15 repetitions of 6 American English vowels in /hVC/ context (hid, head, hat, hot, hub, hoot). Vowel formant frequency measures (F1, F2) were Bark transformed and compared with data from archival sources. RESULTS: Measures of vowel acoustics in speakers with AOS did not differ from those of unimpaired speakers, including absolute Bark formant values, vowel space area, intervowel distance, and individual trial-to-trial formant variability. CONCLUSION: Comparison with normative acoustic measures suggested that vowel production at the word level is unimpaired in the current speakers with AOS, supporting previous studies that have shown vowel production is relatively intact in AOS. PMID- 20008685 TI - Cancer risks and radiation exposure from computed tomographic scans: how can we be sure that the benefits outweigh the risks? PMID- 20008686 TI - A double take on serial measurement of coronary artery calcification. PMID- 20008687 TI - Coffee, decaffeinated coffee, and tea consumption in relation to incident type 2 diabetes mellitus: a systematic review with meta-analysis. AB - BACKGROUND: Coffee consumption has been reported to be inversely associated with risk of type 2 diabetes mellitus. Similar associations have also been reported for decaffeinated coffee and tea. We report herein the findings of meta-analyses for the association between coffee, decaffeinated coffee, and tea consumption with risk of diabetes. METHODS: Relevant studies were identified through search engines using a combined text word and MeSH (Medical Subject Headings) search strategy. Prospective studies that reported an estimate of the association between coffee, decaffeinated coffee, or tea with incident diabetes between 1966 and July 2009. RESULTS: Data from 18 studies with information on 457 922 participants reported on the association between coffee consumption and diabetes. Six (N = 225 516) and 7 studies (N = 286 701) also reported estimates of the association between decaffeinated coffee and tea with diabetes, respectively. We found an inverse log-linear relationship between coffee consumption and subsequent risk of diabetes such that every additional cup of coffee consumed in a day was associated with a 7% reduction in the excess risk of diabetes relative risk, 0.93 [95% confidence interval, 0.91-0.95]) after adjustment for potential confounders. CONCLUSIONS: Owing to the presence of small-study bias, our results may represent an overestimate of the true magnitude of the association. Similar significant and inverse associations were observed with decaffeinated coffee and tea and risk of incident diabetes. High intakes of coffee, decaffeinated coffee, and tea are associated with reduced risk of diabetes. The putative protective effects of these beverages warrant further investigation in randomized trials. PMID- 20008688 TI - Annual progression of coronary calcification in trials of preventive therapies: a systematic review. AB - BACKGROUND: Coronary artery calcification (CAC) measured by computed tomography is radiographic confirmation of atherosclerosis, predicts cardiovascular events, and has been evaluated as a surrogate measure in randomized trials. METHODS: We performed a literature search for prospective randomized trials in which CAC was measured at baseline and at 1 year or more of follow-up. We computed the weighted mean annualized rate of CAC progression for a variety of therapies tested in these trials. RESULTS: Ten trials (n = 2612) met our criteria and were included. Electron-beam, double-helix, and multislice computed tomography were used in 6, 2, and 2 trials, respectively. Agatston (8 trials) and volumetric (2 trials) methods were used for CAC evaluation. In 5 trials in subjects with cardiovascular disease (CVD) (n = 2135; age, ~64 years; ~39% women; follow-up, ~26 months), therapies included statins (n = 1370), placebo (n = 564), and antihypertensives (n = 201). In 5 trials in subjects with chronic kidney disease (n = 477; age, ~55 years; ~34% women; follow-up, ~14 months), interventions included low-phosphorus diet (n = 29), sevelamer hydrochloride (n = 229), and calcium-based phosphate binders (n = 219). The mean (SD) weighted annualized CAC increase overall and in patients with CVD and chronic kidney disease was 17.2% (6.7%), 16.9% (5.2%), and 18.4 (11.1%), respectively (P < .001). The rate among those assigned blinded placebo was 14.6% (1.0%) (2 trials). There was no consistent or reproducible treatment effect of any therapy on this outcome measured at 1 year. CONCLUSION: The 1-year change in CAC does not appear to be a suitable surrogate end point for treatment trials in patients with CVD or chronic kidney disease. PMID- 20008689 TI - Projected cancer risks from computed tomographic scans performed in the United States in 2007. AB - BACKGROUND: The use of computed tomographic (CT) scans in the United States (US) has increased more than 3-fold since 1993 to approximately 70 million scans annually. Despite the great medical benefits, there is concern about the potential radiation-related cancer risk. We conducted detailed estimates of the future cancer risks from current CT scan use in the US according to age, sex, and scan type. METHODS: Risk models based on the National Research Council's "Biological Effects of Ionizing Radiation" report and organ-specific radiation doses derived from a national survey were used to estimate age-specific cancer risks for each scan type. These models were combined with age- and sex-specific scan frequencies for the US in 2007 obtained from survey and insurance claims data. We estimated the mean number of radiation-related incident cancers with 95% uncertainty limits (UL) using Monte Carlo simulations. RESULTS: Overall, we estimated that approximately 29 000 (95% UL, 15 000-45 000) future cancers could be related to CT scans performed in the US in 2007. The largest contributions were from scans of the abdomen and pelvis (n = 14 000) (95% UL, 6900-25 000), chest (n = 4100) (95% UL, 1900-8100), and head (n = 4000) (95% UL, 1100-8700), as well as from chest CT angiography (n = 2700) (95% UL, 1300-5000). One-third of the projected cancers were due to scans performed at the ages of 35 to 54 years compared with 15% due to scans performed at ages younger than 18 years, and 66% were in females. CONCLUSIONS: These detailed estimates highlight several areas of CT scan use that make large contributions to the total cancer risk, including several scan types and age groups with a high frequency of use or scans involving relatively high doses, in which risk-reduction efforts may be warranted. PMID- 20008690 TI - Radiation dose associated with common computed tomography examinations and the associated lifetime attributable risk of cancer. AB - BACKGROUND: Use of computed tomography (CT) for diagnostic evaluation has increased dramatically over the past 2 decades. Even though CT is associated with substantially higher radiation exposure than conventional radiography, typical doses are not known. We sought to estimate the radiation dose associated with common CT studies in clinical practice and quantify the potential cancer risk associated with these examinations. METHODS: We conducted a retrospective cross sectional study describing radiation dose associated with the 11 most common types of diagnostic CT studies performed on 1119 consecutive adult patients at 4 San Francisco Bay Area institutions in California between January 1 and May 30, 2008. We estimated lifetime attributable risks of cancer by study type from these measured doses. RESULTS: Radiation doses varied significantly between the different types of CT studies. The overall median effective doses ranged from 2 millisieverts (mSv) for a routine head CT scan to 31 mSv for a multiphase abdomen and pelvis CT scan. Within each type of CT study, effective dose varied significantly within and across institutions, with a mean 13-fold variation between the highest and lowest dose for each study type. The estimated number of CT scans that will lead to the development of a cancer varied widely depending on the specific type of CT examination and the patient's age and sex. An estimated 1 in 270 women who underwent CT coronary angiography at age 40 years will develop cancer from that CT scan (1 in 600 men), compared with an estimated 1 in 8100 women who had a routine head CT scan at the same age (1 in 11 080 men). For 20 year-old patients, the risks were approximately doubled, and for 60-year-old patients, they were approximately 50% lower. CONCLUSION: Radiation doses from commonly performed diagnostic CT examinations are higher and more variable than generally quoted, highlighting the need for greater standardization across institutions. PMID- 20008692 TI - Should there be a fracas over FRAX and other fracture prediction tools?: Comment on "A comparison of prediction models for fractures in older women". PMID- 20008691 TI - A comparison of prediction models for fractures in older women: is more better? AB - BACKGROUND: A Web-based risk assessment tool (FRAX) using clinical risk factors with and without femoral neck bone mineral density (BMD) has been incorporated into clinical guidelines regarding treatment to prevent fractures. However, it is uncertain whether prediction with FRAX models is superior to that based on parsimonious models. METHODS: We conducted a prospective cohort study in 6252 women 65 years or older to compare the value of FRAX models that include BMD with that of parsimonious models based on age and BMD alone for prediction of fractures. We also compared FRAX models without BMD with simple models based on age and fracture history alone. Fractures (hip, major osteoporotic [hip, clinical vertebral, wrist, or humerus], and any clinical fracture) were ascertained during 10 years of follow-up. Area under the curve (AUC) statistics from receiver operating characteristic curve analysis were compared between FRAX models and simple models. RESULTS: The AUC comparisons showed no differences between FRAX models with BMD and simple models with age and BMD alone in discriminating hip (AUC, 0.75 for the FRAX model and 0.76 for the simple model; P = .26), major osteoporotic (AUC, 0.68 for the FRAX model and 0.69 for the simple model; P = .51), and clinical fracture (AUC, 0.64 for the FRAX model and 0.63 for the simple model; P = .16). Similarly, performance of parsimonious models containing age and fracture history alone was nearly identical to that of FRAX models without BMD. The proportion of women in each quartile of predicted risk who actually experienced a fracture outcome did not differ between FRAX and simple models (P > or = .16). CONCLUSION: Simple models based on age and BMD alone or age and fracture history alone predicted 10-year risk of hip, major osteoporotic, and clinical fracture as well as more complex FRAX models. PMID- 20008693 TI - Combined effects of cardiorespiratory fitness, not smoking, and normal waist girth on morbidity and mortality in men. AB - BACKGROUND: Physical inactivity, cigarette smoking, and abdominal obesity are key modifiable risk factors for coronary heart disease (CHD). We investigated the combined effects of not having these risk factors on CHD events and cardiovascular disease (CVD) and all-cause mortality in men. METHODS: We followed up 23 657 men, aged 30 to 79 years, who completed a medical evaluation including a maximal treadmill exercise test and self-reported health habits. A low-risk profile was defined as not smoking, moderate or high fitness, and normal waist girth. There were 482 CHD events (nonfatal myocardial infarction or fatal CHD) and 1034 deaths (306 CVD, 387 cancers, and 341 others) during a mean 14.7 years of follow-up (348 811 man-years). RESULTS: After adjustment for age, examination year, and multiple baseline risk factors, there was an inverse association between a greater number of low-risk factors and CHD events, and CVD and all cause mortality in men (P value for trend, <.001 for all). Men with a normal waist girth and who were physically fit and not smoking had a 59% lower risk of CHD events (95% confidence interval [CI], 39%-72%), a 77% lower risk of CVD mortality (95% CI, 65%-85%), and a 69% lower risk of all-cause mortality (95% CI, 60%-76%) compared with men with none of these low-risk factors. Men with 0 compared with 3 low-risk factors had a shorter life expectancy by 14.2 years (95% CI, 12.2-15.9 years). CONCLUSION: Being physically fit, not smoking, and maintaining a normal waist girth is associated with lower risk of CHD events, and CVD and all-cause mortality in men. PMID- 20008695 TI - Effects of television viewing reduction on energy intake and expenditure in overweight and obese adults: a randomized controlled trial. AB - BACKGROUND: The average adult watches almost 5 hours of television (TV) per day, an amount associated with increased risks for obesity. This trial examines the effects of TV reduction on energy intake (EI), energy expenditure (EE), energy balance, body mass index (BMI), (calculated as weight in kilograms divided by height in meters squared), and sleep in overweight and obese adults. METHODS: Randomized controlled trial of 36 adults with a BMI of 25 to 50 who self-reported a minimum of 3 h/d of TV viewing. Participants were enrolled in home-based protocols from January through July 2008. After a 3-week observation phase, participants were stratified by BMI and randomized to an observation-only control group (n = 16) or an intervention group (n = 20) for 3 additional weeks. The intervention consisted of reducing TV viewing by 50% of each participant's objectively measured baseline enforced by an electronic lock-out system. RESULTS: Although not statistically significant, both groups reduced their EI (-125 kcal/d [95% CI, -303 to 52] vs -38 [95% CI, -265 to 190]) (P = .52) for intervention and control group participants, respectively, where CI indicates confidence interval. The intervention group significantly increased EE (119 kcal/d [95% CI, 23 to 215]) compared with controls (-95 kcal/d [95% CI, -254 to 65]) (P = .02). Energy balance was negative in the intervention group between phases (-244 kcal/d [95% CI, -459 to -30]) but positive in controls (57 kcal/d [95% CI, -216 to 330]) (P = .07). The intervention group showed a greater reduction in BMI (-0.25 [95% CI, 0.45 to -0.05] vs -0.06 [95% CI, -0.43 to 0.31] in controls) (P = .33). There was no change in sleep. CONCLUSION: Reducing TV viewing in our sample produced a statistically significant increase in EE but no apparent change in EI after 3 weeks of intervention. Trial Registration clinicaltrials.gov Identifier: NCT00622050. PMID- 20008694 TI - Physical activity and male colorectal cancer survival. AB - BACKGROUND: Although physically active individuals have a lower risk of developing colorectal cancer, few studies have examined whether exercise benefits colorectal cancer survivors. METHODS: Derived from the Health Professionals Follow-up Study, we studied colorectal cancer-specific and overall mortality in a cohort of 668 men with a history of stage I to stage III colorectal cancer according to predefined physical activity categories after diagnosis. To minimize bias by occult recurrences, we excluded men who died within 6 months of their postdiagnosis physical activity assessment. RESULTS: In a cohort of men with colorectal cancer and no apparent metastases at diagnosis, 50.4% exercised at least 18 metabolic equivalent task (MET) hours per week. Increased physical activity was significantly associated with improved colorectal cancer-specific mortality (P = .002 for trend) and overall mortality (P < .001 for trend). Men who engaged in more than 27 MET hours per week of physical activity had an adjusted hazard ratio for colorectal cancer-specific mortality of 0.47 (95% confidence interval, 0.24-0.92) compared with men who engaged in 3 or less MET hours per week of physical activity. The apparent benefit of physical activity was seen regardless of age, disease stage, body mass index, diagnosis year, tumor location, and prediagnosis physical activity. CONCLUSION: In a large cohort of men with a history of nonmetastatic colorectal cancer, more physical activity was associated with a lower risk of colorectal cancer-specific and overall mortality. PMID- 20008696 TI - Physical activity and rapid decline in kidney function among older adults. AB - BACKGROUND: Habitual physical activity (PA) has both physiologic and metabolic effects that may moderate the risk of kidney function decline. We tested the hypothesis that higher levels of PA are associated with a lower risk of kidney function decline using longitudinal data from a large cohort of older adults. METHODS: We studied 4011 ambulatory participants aged 65 or older from the Cardiovascular Health Study (CHS) who completed at least 2 measurements of kidney function over 7 years. We calculated a PA score (range, 2-8) by summing kilocalories expended per week (ordinal score of 1-5 from quintiles of kilocalories per week) and walking pace (ordinal score for categories of <2, 2-3, and >3 mph). Rapid decline in kidney function decline (RDKF) was defined by loss of more than 3.0 mL/min/1.73 m(2) per year in glomerular filtration rate, which we estimated by using longitudinal measurements of cystatin C levels. RESULTS: A total of 958 participants had RDKF (23.9%; 4.1 events per 100 person-years). The estimated risk of RDKF was 16% in the highest PA group (score of 8) and 30% in the lowest PA group (score of 2). After multivariate adjustment, we found that the 2 highest PA groups (scores of 7-8) were associated with a 28% lower risk of RDKF (95% confidence interval, 21%-41% lower risk) than the 2 lowest PA groups (score of 2-3). Greater kilocalories of leisure-time PA and walking pace were also each associated with a lower incidence of RDKF. CONCLUSION: Higher levels of PA are associated with a lower risk of RDKF among older adults. PMID- 20008697 TI - Physical activity: an investment that pays multiple health dividends: comment on "combined effects of cardiorespiratory fitness, not smoking, and normal waist girth on morbidity and mortality in men," "physical activity and survival in male colorectal cancer survival," "effects of a television viewing reduction on energy intake and expenditure in overweight and obese adults," and "physical activity and rapid decline in kidney function among older adults". PMID- 20008698 TI - Antidepressant use and risk of incident cardiovascular morbidity and mortality among postmenopausal women in the Women's Health Initiative study. AB - BACKGROUND: Antidepressants are commonly prescribed medications, but their effect on cardiovascular morbidity and mortality remains unclear. METHODS: Prospective cohort study of 136 293 community-dwelling postmenopausal women in the Women's Health Initiative (WHI). Women taking no antidepressants at study entry and who had at least 1 follow-up visit were included. Cardiovascular morbidity and all cause mortality for women with new antidepressant use at follow-up (n = 5496) were compared with those characteristics for women taking no antidepressants at follow-up (mean follow-up, 5.9 years). RESULTS: Antidepressant use was not associated with coronary heart disease (CHD). Selective serotonin reuptake inhibitor (SSRI) use was associated with increased stroke risk (hazard ratio [HR],1.45, [95% CI, 1.08-1.97]) and all-cause mortality (HR,1.32 [95% CI, 1.10 1.59]). Annualized rates per 1000 person-years of stroke with no antidepressant use and SSRI use were 2.99 and 4.16, respectively, and death rates were 7.79 and 12.77. Tricyclic antidepressant (TCA) use was associated with increased risk of all-cause mortality (HR,1.67 [95% CI, 1.33-2.09]; annualized rate, 14.14 deaths per 1000 person-years). There were no significant differences between SSRI and TCA use in risk of any outcomes. In analyses by stroke type, SSRI use was associated with incident hemorrhagic stroke (HR, 2.12 [95% CI, 1.10-4.07]) and fatal stroke (HR, 2.10 [95% CI, 1.15-3.81]). CONCLUSIONS: In postmenopausal women, there were no significant differences between SSRI and TCA use in risk of CHD, stroke, or mortality. Antidepressants were not associated with risk of CHD. Tricyclic antidepressants and SSRIs may be associated with increased risk of mortality, and SSRIs with increased risk of hemorrhagic and fatal stroke, although absolute event risks are low. These findings must be weighed against quality of life and established risks of cardiovascular disease and mortality associated with untreated depression. PMID- 20008699 TI - Antidepressant Use, Depression, and Poor Cardiovascular Outcomes: The Chicken or the Egg?: Comment on "Antidepressant Use and Risk of Incident Cardiovascular Morbidity and Mortality Among Postmenopausal Women in the Women's Health Initiative Study". PMID- 20008700 TI - Mental morbidities and chronic fatigue in severe acute respiratory syndrome survivors: long-term follow-up. AB - BACKGROUND: Short-term follow-up studies of severe acute respiratory syndrome (SARS) survivors suggested that their physical conditions continuously improved in the first year but that their mental health did not. We investigated long-term psychiatric morbidities and chronic fatigue among SARS survivors. METHODS: All SARS survivors from the hospitals of a local region in Hong Kong were assessed by a constellation of psychometric questionnaires and a semistructured clinical interview for the Diagnostic and Statistical Manual of Mental Disorders (Fourth Edition) to determine the presence of psychiatric disorders and chronic fatigue problems. RESULTS: Of 369 SARS survivors, 233 (63.1%) participated in the study (mean period of time after SARS, 41.3 months). Over 40% of the respondents had active psychiatric illnesses, 40.3% reported a chronic fatigue problem, and 27.1% met the modified 1994 Centers for Disease Control and Prevention criteria for chronic fatigue syndrome. Logistic regression analysis suggested that being a health care worker at the time of SARS infection (odds ratio [OR], 3.24; 95% confidence interval [CI], 1.12- 9.39; P = .03), being unemployed at follow-up (OR, 4.71; 95% CI, 1.50-14.78; P = .008), having a perception of social stigmatization (OR, 3.03; 95% CI, 1.20-7.60; P = .02), and having applied to the SARS survivors' fund (OR, 2.92; 95% CI, 1.18-7.22; P = .02) were associated with an increased risk of psychiatric morbidities at follow-up, whereas application to the SARS survivors' fund (OR, 2.64; 95% CI, 1.07-6.51; P = .04) was associated with increased risk of chronic fatigue problems. CONCLUSIONS: Psychiatric morbidities and chronic fatigue persisted and continued to be clinically significant among the survivors at the 4-year follow-up. Optimization of the treatment of mental health morbidities by a multidisciplinary approach with a view for long-term rehabilitation, especially targeting psychiatric and fatigue problems and functional and occupational rehabilitation, would be needed. PMID- 20008701 TI - Comparative effectiveness of 5 smoking cessation pharmacotherapies in primary care clinics. AB - BACKGROUND: Randomized efficacy clinical trials conducted in research settings may not accurately reflect the benefits of tobacco dependence treatments when used in real-world clinical settings. Effectiveness trials (eg, in primary care settings) are needed to estimate the benefits of cessation treatments in real world use. METHODS: A total of 1346 primary care patients attending routine appointments were recruited by medical assistants in 12 primary care clinics. Patients were randomly assigned to 5 active pharmacotherapies: 3 monotherapies (nicotine patch, nicotine lozenge, and bupropion hydrochloride sustained release [SR]) and 2 combination therapies (patch + lozenge and bupropion SR + lozenge). Patients were referred to a telephone quit line for cessation counseling. Primary outcomes included 7-day point prevalence abstinence at 1 week, 8 weeks, and 6 months after quitting and number of days to relapse. RESULTS: Among 7128 eligible smokers (> or =10 cigarettes per day) attending routine primary care appointments, 1346 (18.9%) were enrolled in the study. Six-month abstinence rates for the 5 active pharmacotherapies were the following: bupropion SR, 16.8%; lozenge, 19.9%; patch, 17.7%; patch + lozenge, 26.9%; and bupropion SR + lozenge, 29.9%. Bupropion SR + lozenge was superior to all of the monotherapies (odds ratio, 0.46-0.56); patch + lozenge was superior to patch and bupropion monotherapies (odds ratio, 0.56 and 0.54, respectively). CONCLUSIONS: One in 5 smokers attending a routine primary care appointment was willing to make a serious quit attempt that included evidence-based counseling and medication. In this comparative effectiveness study of 5 tobacco dependence treatments, combination pharmacotherapy significantly increased abstinence compared with monotherapies. Provision of free cessation medications plus quit line counseling arranged in the primary care setting holds promise for assisting large numbers of smokers to quit. Trial Registration clinicaltrials.gov Identifier: NCT00296647. PMID- 20008702 TI - Spouse-rated vs self-rated health as predictors of mortality. AB - BACKGROUND: The Health and Retirement Study is a national sample of Americans older than 50 years and their spouses. The present study evaluated cross sectional and longitudinal data from January 2000 through December 2006. The objective of the study was to evaluate the roles of spouse-rated vs self-rated health as predictors of all-cause mortality among adults older than 50 years. METHODS: A total of 673 dyads of married couples were randomly selected to participate in a Health and Retirement Study module examining spouse-rated health. For each couple, one member was asked to rate his or her overall health status, and his or her spouse was asked to report the partner's overall health status. Mortality data were available through 2006. RESULTS: Our findings demonstrate that spouse-rated health (area under the curve, 0.75) is as strong a predictor of mortality as self-rated health (area under the curve, 0.73) (chi(2)(1) = 0.36, P = .54). Combining spouse-rated and self-rated health predicts mortality better than using self-rated health alone (area under the curve, 0.77) (chi(2)(1) = 6.72, P = .009). CONCLUSIONS: Spouse ratings of health are at least as strongly predictive of mortality as self-rated health. This suggests that, when self-rated health is elicited as a prognostic indicator, spouse ratings can be used when self-ratings are unavailable. Both measures together may be more informative than either measure alone. PMID- 20008703 TI - Discontinuation of screening mammography after serious health events. PMID- 20008704 TI - It is time to get more accurate times to defibrillation. PMID- 20008705 TI - Antipsychotic drug-associated acute hyperglycemia in elderly diabetic patients: do the drugs explain the hyperglycemia? PMID- 20008706 TI - Antipsychotic use and risk for hyperglycemia in elderly patients with DM. PMID- 20008707 TI - Carotid disease and syncope. PMID- 20008708 TI - Exploratory analysis of diabetic retinopathy progression through 3 years in a randomized clinical trial that compares intravitreal triamcinolone acetonide with focal/grid photocoagulation. AB - OBJECTIVE: To compare the effect of intravitreal triamcinolone acetonide with focal/grid photocoagulation on the progression of diabetic retinopathy. METHODS: We performed an exploratory analysis of participants with diabetic macular edema randomly assigned to receive laser therapy or intravitreal triamcinolone acetonide (1 or 4 mg). Fundus photographs were obtained at baseline and 1, 2, and 3 years. The main outcome measure was progression to proliferative diabetic retinopathy or worsening of 2 or more severity levels on reading-center masked assessment of 7-field fundus photographs, plus additional eyes that received panretinal photocoagulation or had a vitreous hemorrhage. RESULTS: From July 15, 2004, through May 5, 2006, 840 eyes from 693 participants were enrolled in the study and randomly assigned to receive laser therapy (n = 330), 1 mg of triamcinolone acetonide (n = 256), or 4 mg of triamcinolone acetonide (n = 254). The cumulative probability of progression of retinopathy at 2 years was 31% (laser group), 29% (1-mg group), and 21% (4-mg group) (P = .64 in the 1-mg group and .005 in the 4-mg group compared with the laser group). These differences appeared to be sustained at 3 years. CONCLUSIONS: Intravitreal triamcinolone acetonide (4 mg) appeared to reduce the risk of progression of diabetic retinopathy. Given the exploratory nature of this analysis and because intravitreal triamcinolone adverse effects include cataract formation and glaucoma, use of this treatment merely to reduce the rates of progression of proliferative diabetic retinopathy or worsening of the level of diabetic retinopathy does not seem warranted at this time. PMID- 20008709 TI - Melanoma-associated retinopathy: a paraneoplastic autoimmune complication. AB - OBJECTIVES: To study 11 patients with melanoma-associated retinopathy (MAR) to clarify the reliability of various methods of diagnostic testing, to determine the underlying antigenic retinal proteins, and to study the clinical histories and types of associated melanomas. METHODS: Clinical data were obtained from patients with melanoma who developed marked visual problems. Testing included electroretinography, kinetic visual fields, comparative studies of Western blots, and indirect immunohistologic examination to detect antiretinal antibodies, as well as proteomic studies to identify underlying antigenic retinal proteins. RESULTS: Patients with MAR typically have rapid onset of photopsias, scotomata, and loss of central or paracentral vision. Ophthalmoscopy seldom shows significant changes early, but electroretinograms are abnormal. Results of Western blots and immunohistologic examination can show antiretinal antibodies but not always. Most patients (9 of 11) had a strong family history of autoimmune disorders. Any type of melanoma (cutaneous, choroidal, ciliary body, or choroidal nevi) may be associated with this paraneoplastic autoimmune reactivity. MAR may precede or follow the diagnosis of melanoma. Patients with MAR have the same antigenic retinal proteins that have been associated with cancer-associated retinopathy. In addition, 2 new antigenic retinal proteins, aldolase A and aldolase C, were found. CONCLUSIONS: There was a high prevalence of positive family histories of autoimmune disease in patients with MAR. To confirm the disorder, multiple clinical and serum diagnostic techniques (Western blot or indirect immunohistologic examination) are needed. Two newly observed antigenic retinal proteins, aldolase A and aldolase C, are associated with MAR. PMID- 20008711 TI - Short tag noose technique for optional and late suture adjustment in strabismus surgery. AB - OBJECTIVE: To present and evaluate a new technique that allows the second-stage suture adjustment in strabismus surgery to be skipped or delayed if the immediate postoperative alignment is satisfactory. METHODS: The "short tag noose" technique replaces long sutures with short tags that can be left under the conjunctiva after adjustment. Retrospectively, the medical records of all patients treated by a single surgeon with this approach between January 1, 2005, and December 31, 2008, were evaluated for success rate (< or =10 prism diopters [PD] horizontal and < or =6 PD vertical), reoperation rate, and complications. RESULTS: Of 120 procedures reviewed, 27 (22.5%) were performed in children; 97 procedures (80.8%) were performed in patients with complex strabismus. Mean follow-up was 6 months. The success rate was 81.0% for horizontal strabismus and 70.7% for vertical strabismus at 2 months. The reoperation rate was 10.0% for horizontal strabismus and 19.0% for vertical strabismus. The noose slipped in 1 patient (0.8%) and was corrected by readjustment on day 4. Two patients (1.7%) required in-office excision of cysts or granulomas. CONCLUSIONS: The short tag noose technique simplifies the logistics of suture adjustment and avoids the need for sedation in children who do not require adjustment. It provides the ability to defer adjustment for days after surgery. PMID- 20008710 TI - Risk of endophthalmitis after intravitreal drug injection when topical antibiotics are not required: the diabetic retinopathy clinical research network laser-ranibizumab-triamcinolone clinical trials. AB - OBJECTIVE: To report the incidence of endophthalmitis after intravitreal drug injection by means of a standardized procedure that does not require topical antibiotics, sterile gloves, or a sterile drape. METHODS: Intravitreal injections of preservative-free triamcinolone acetonide or ranibizumab were administered in 2 prospective randomized clinical trials performed by the Diabetic Retinopathy Clinical Research Network. The standardized procedure for these trials requires the use of a topical combination product of povidone-iodine, a sterile lid speculum, and topical anesthetic, but does not require the use of topical antibiotics before, on the day of, or after injection. RESULTS: As of February 23, 2009, a total of 3226 intravitreal injections of ranibizumab and 612 injections of preservative-free triamcinolone had been administered. Topical antibiotics were given on the day of injection in 361 (9.4%) of the 3838 cases, for several days after injection in 813 cases (21.2%), on the day of injection and after injection in 1388 cases (36.2%), and neither on the day of injection nor after injection in 1276 cases (33.3%). Three cases of culture-positive endophthalmitis occurred after ranibizumab injections (0.09%), and no cases occurred after triamcinolone injections. In all 3 cases of endophthalmitis, topical antibiotics were given for several days after the injection but not before injection. CONCLUSIONS: The results suggest that a low rate of endophthalmitis can be achieved by means of a protocol that includes use of topical povidone-iodine, a sterile lid speculum, and topical anesthetic, but does not require topical antibiotics, sterile gloves, or a sterile drape. Trial Registration clinicaltrials.gov Identifiers: NCT00444600 and NCT00445003. PMID- 20008712 TI - Dr Thompson's Eye Water. PMID- 20008713 TI - Comparison between the plusoptiX and MTI Photoscreeners. AB - OBJECTIVE: Both the Medical Technology and Innovations (MTI) and plusoptiX photoscreeners are used to objectively screen for amblyogenic risk factors in children. The MTI has been extensively studied, but the limited availability of film may render it obsolete. We compared the MTI with the plusoptiX, a newer digital photoscreener, for the ability to detect amblyogenic factors when compared with a comprehensive pediatric ophthalmic examination. We believe our results will help to guide community-based vision screening programs. METHODS: One hundred fifty-one children were examined consecutively in our office. Each patient was screened with the MTI and plusoptiX devices on the same day as part of a comprehensive pediatric ophthalmic examination. Results via MTI were evaluated by an expert masked examiner (R.W.A.), and the plusoptiX results were interpreted by the incorporated software. RESULTS: Sixty-five percent of patients were found to have amblyopia or amblyogenic risk factors during the pediatric ophthalmic examination conducted via the American Association of Pediatric Ophthalmology and Strabismus referral criteria. We found the MTI photoscreener to have a sensitivity of 83.6%, specificity of 90.5%, false- positive rate of 9.4%, false-negative rate of 16.3%, and positive predictive value of 94.2%. The plusoptiX demonstrated a sensitivity of 98.9%, specificity of 96.1%, false- positive rate of 3.7%, false-negative rate of 1.0%, and positive predictive value of 97.9%. CONCLUSION: The MTI and plusoptiX photoscreeners proved to be effective when compared with a comprehensive cycloplegic pediatric ophthalmic examination. The plusoptiX, however, was found to have a higher sensitivity and specificity than the MTI. PMID- 20008714 TI - Outer retinal tubulation: a novel optical coherence tomography finding. AB - OBJECTIVE: To describe tubular structures found in the outer retina seen in a variety of retinal disorders. METHODS: Sixty-nine eyes of 63 patients were examined with spectral-domain optical coherence tomography. Optical coherence tomography C-scans were correlated with their corresponding B-scans. The prevalence, number, size, and shape of the tubular structures were determined. RESULTS: Branching tubules were identified in the outer retina of 54 patients with age-related macular degeneration and in 9 patients with other diagnoses. The tubules appeared as round or ovoid hyporeflective spaces with hyperreflective borders on the B-scans, measuring 40 to 140 microm high and 40 to 2260 microm wide. Morphologic features ranged from single straight or branching tubules to complex cavitary networks, usually overlying areas of pigment epithelial alteration or subretinal fibrosis. The tubules generally remained stable over time. In a retinal practice specializing in advanced age-related macular degeneration, these structures were identified in 60 of 248 patients (24.2%) seen during a 3-month period. CONCLUSIONS: Degenerating photoreceptors may become arranged in a circular or ovoid fashion during a process we propose to term outer retinal tubulation. These changes are apparently common in advanced diseases affecting the outer retina and retinal pigment epithelium. This observation has practical implications because these findings can be misinterpreted as intraretinal or subretinal fluid, possibly prompting unnecessary interventions. PMID- 20008715 TI - Comparison of glaucoma diagnostic Capabilities of Cirrus HD and Stratus optical coherence tomography. AB - OBJECTIVE: To compare the glaucoma diagnostic capabilities offered by Stratus and Cirrus spectral-domain optical coherence tomography (OCT). METHODS: One hundred subjects with glaucoma and 74 healthy subjects were tested by Stratus and Cirrus OCT. Areas under the receiver operating characteristic curves (AUCs) of average, 4-quadrant, and 12-sector retinal nerve fiber layer thicknesses and sensitivities at fixed specificities (80% and 90%) were compared when the 2 OCT modalities were used to evaluate patients with early or moderate to advanced glaucoma. Likelihood ratios using normative classifications were reported. RESULTS: Overall, both OCT instruments showed similar glaucoma discrimination capability in average retinal nerve fiber layer thickness (AUC, 0.953 [Cirrus] vs 0.934 [Stratus]; P = .15). Cirrus OCT displayed significantly higher AUCs in the average, inferior, temporal, and superior quadrants and 7-o'clock measurements in early stages of glaucoma. The between-OCT instrument AUCs did not differ significantly in moderate to advanced stages. Abnormal results for both OCT instruments, after comparison with their normative databases, were associated with high likelihood ratios. CONCLUSIONS: In our series, the Cirrus OCT showed better glaucoma discrimination capability than Stratus OCT in early stages of glaucoma. Our findings suggest that spectral-domain technology of OCT may offer an improved capability of early-stage glaucoma detection. PMID- 20008716 TI - Prediction of glaucomatous visual field loss by extrapolation of linear trends. AB - OBJECTIVE: To investigate how well short-term progression rates can predict long term visual field outcomes in patients with glaucoma. METHODS: We calculated visual field rates of progression using linear regression analysis of the Visual Field Index (VFI) for 100 consecutive patients with glaucoma having 10 or more Swedish Interactive Thresholding Algorithm standard field tests. Final VFI was predicted on the basis of linear extrapolation of the slope defined by the initial 5 field test results. Final VFI also was estimated using linear regression of all qualifying examination results for each patient. Primary outcome measures were the absolute difference and the correlation between predicted and estimated final VFI values. RESULTS: Patient follow-up averaged 8.2 years and 11 field examinations. Median VFI progression rate was -1.1% per year both for the initial 5 test results and also for the complete series. Seventy percent of patients had a predicted final VFI within +/-10% of the estimated final VFI, and the 2 VFI calculations had a correlation coefficient of 0.84. CONCLUSION: Linear extrapolation based on 5 initial visual field test results was a reliable predictor of future field loss in most patients. Patients in whom linear regression analysis suggests dangerously rapid rates of visual field progression may be candidates for significant alterations in therapy. PMID- 20008717 TI - Contact lens-related adverse events and the silicone hydrogel lenses and daily wear care system used. AB - OBJECTIVE: To investigate the incidence of adverse events related to the use of varying silicone hydrogel contact lens and lens solution combinations. METHODS: Individuals with myopia (N = 558) participated in 1 or more of approximately 40 participant trials in a matrix of 20 silicone hydrogel contact lens and lens solution combinations. Visits were at baseline, 2 weeks, 1 month, and 3 months. The mean study completion rate was 90% of the expected participant-months (final data set: 840 lens-solution combinations and 2271 participant-months). Adverse events were reported as the first occurrence of each type per 100 participant months for each lens-solution combination. RESULTS: The rate of all corneal infiltrative events (CIEs) was 3.1 per 100 participant-months (range, 0-10.5), and the rate of symptomatic CIEs was 1.7 per 100 participant-months (range, 0 10.5), including 1 case of microbial keratitis (0.04 per 100 participant-months). Rates for CIEs differed substantially among solution groups, with hydrogen peroxide having the lowest rate (0.6 per 100 participant-months; range, 0-0.9). The rate was 0.8 per 100 participant-months (range, 0-8.0) for superior epithelial arcuate lesions, which varied by lens type, 0.04 per 100 participant months (1 case only) for corneal erosion, and 0.4 per 100 participant-months (range, 0-2.0) for contact lens papillary conjunctivitis, which was modified by type of solution. The rate of solution-induced corneal staining for all lens solution combinations was 4.7 per 100 participant-months (range, 0-23) and varied significantly based on lens-solution combination (P < .001). CONCLUSIONS: The frequency of adverse events varied with silicone hydrogel contact lens and lens solution combinations, with hydrogen peroxide having the lowest incidence of CIEs and solution-induced corneal staining, indicating that lens material and design, type of solution, and solution-lens interactions are likely contributing factors in this mode of lens wear. PMID- 20008718 TI - Essential role for c-Jun N-terminal kinase 2 in corneal epithelial response to desiccating stress. AB - OBJECTIVE: To investigate the protective effects of c-Jun N-terminal kinase (JNK) 1 and -2 gene knockout (KO) on the corneal epithelial response to desiccating stress. METHODS: The C57BL/6, JNK1KO, and JNK2KO mice were subjected to desiccating stress (DS) for 5 days. The effects of DS on the corneal epithelium were evaluated by measuring corneal smoothness and permeability. Expression of matrix metalloproteinases (MMP)-1, MMP-9, and cornified envelope protein precursors (small proline-rich protein [SPRR]-1a, SPRR-2a, and involucrin) in the corneal epithelia was evaluated by immunostaining and real-time polymerase chain reaction. Collagenase and gelatinase activity in corneal sections as measured with in situ fluorescent assays. RESULTS: The JNK2KO mice had smoother corneal surfaces and less corneal barrier disruption in response to DS than JNK1KO mice and C57BL/6 wild-type control mice. The DS increased levels of MMP-1, MMP-9, SPRR 1a, SPRR-2a, involucrin immunoreactivity, and mRNA transcripts in the corneal epithelium of JNK1KO and C57BL/6 mice, but not in JNK2KO mice. Knockout of JNK2 prevented DS-induced increase in gelatinase and collagenase activity in the cornea. CONCLUSION: The JNK2 protein appears to have an essential role in desiccation-induced corneal epithelial disease by stimulating production of MMP 1, MMP-9, and cornified envelope precursors. Clinical Relevance The JNK2 protein could be a novel therapeutic target in dry eye disease. PMID- 20008719 TI - Increased prevalence of myopia in the United States between 1971-1972 and 1999 2004. AB - OBJECTIVE: To compare US population prevalence estimates for myopia in 1971-1972 and 1999-2004. METHODS: The 1971-1972 National Health and Nutrition Examination Survey provided the earliest nationally representative estimates for US myopia prevalence; myopia was diagnosed by an algorithm using either lensometry, pinhole visual acuity, and presenting visual acuity (for presenting visual acuity > or =20/40) or retinoscopy (for presenting visual acuity < or =20/50). Using a similar method for diagnosing myopia, we examined data from the 1999-2004 National Health and Nutrition Examination Survey to determine whether myopia prevalence had changed during the 30 years between the 2 surveys. RESULTS: Using the 1971-1972 method, the estimated prevalence of myopia in persons aged 12 to 54 years was significantly higher in 1999-2004 than in 1971-1972 (41.6% vs 25.0%, respectively; P < .001). Prevalence estimates were higher in 1999-2004 than in 1971-1972 for black individuals (33.5% vs 13.0%, respectively; P < .001) and white individuals (43.0% vs 26.3%, respectively; P < .001) and for all levels of myopia severity (>-2.0 diopters [D]: 17.5% vs 13.4%, respectively [P < .001]; < or =-2.0 to >-7.9 D: 22.4% vs 11.4%, respectively [P < .001]; < or =-7.9 D: 1.6% vs 0.2%, respectively [P < .001]). CONCLUSIONS: When using similar methods for each period, the prevalence of myopia in the United States appears to be substantially higher in 1999-2004 than 30 years earlier. Identifying modifiable risk factors for myopia could lead to the development of cost-effective interventional strategies. PMID- 20008720 TI - Sociodemographic, lifestyle, and medical risk factors for visual impairment in an urban asian population: the singapore malay eye study. AB - OBJECTIVE: To describe the associations between sociodemographic, lifestyle, and medical risk factors and visual impairment in a Southeast Asian population. METHODS: Population-based cross-sectional study of 3280 (78.7% response rate) Malay Singaporeans aged 40 to 80 years. Participants underwent a standardized interview, in which detailed sociodemographic histories were obtained, and clinical assessments for presenting and best-corrected visual acuity. Visual impairment (logMAR > 0.30) was classified as unilateral (1 eye impaired) or bilateral (both eyes impaired). Analyses used multivariate-adjusted multinomial logistic regression. RESULTS: Older age and lack of formal education was associated with increased odds of both unilateral and bilateral visual impairment based on presenting and best-corrected visual acuity. The odds doubled for each decade older, and lower education increased the odds 1.59- to 2.83-fold. Bilateral visual impairment was associated with being unemployed (odds ratio [OR], 1.84; 95% confidence interval [CI], 1.30-2.60), widowed status (OR, 1.51; 95% CI, 1.13-2.01), and higher systolic blood pressure (OR, 1.96; 95% CI, 1.44 2.66). Diabetes was associated with unilateral (OR, 1.47; 95% CI, 1.10-1.95) and bilateral (OR, 1.69; 95% CI, 1.23-2.32) visual impairment using best-corrected visual acuity. CONCLUSIONS: Older age, lower education, unemployment, being widowed, diabetes, and hypertension were independently associated with bilateral visual impairment. Public health interventions should be targeted to these at risk populations. PMID- 20008721 TI - Clinical presentation and genetic correlation of patients with mutations affecting the FZD4 gene. AB - OBJECTIVE: To correlate the ophthalmic findings of patients with pediatric vitreoretinopathies with mutations occurring in the FZD4 gene. METHODS: A total of 123 patients diagnosed with autosomal-dominant familial exudative vitreoretinopathy (AdFEVR) or retinopathy of prematurity (ROP) and 42 control patients were enrolled in the study. Diagnoses were based on retinal findings at each patient's first examination or during ROP screening. Genomic DNA was isolated and polymerase chain reaction and direct sequencing of the FZD4 gene performed. RESULTS: FZD4 gene mutations were discovered in 13 of the 123 (10.6%) patients. Nine of the 63 patients with AdFEVR (14.3%) has mutations in the FZD4 gene. Four heterozygous mutations were identified: C117R, C181Y, Q505X, and P33S/P168S. Four of the 60 patients with ROP (6.7%) have a double missense mutation P33S/P168S that was also found in the patients with FEVR. No other FZD4 mutations were found in the patients with ROP. Additionally, patients expressing the double mutation had clinical presentations that overlapped, making it difficult to assign a definitive diagnosis. None of the mutations found in the patients with FEVR or ROP were seen in the control chromosomes. CONCLUSION: Mutations occurring in the FZD4 gene affect patients diagnosed with both FEVR and ROP. The clinical picture often overlaps and may require a detailed birth and family history for diagnosis. Genetic testing confirms inherited vitreoretinopathy and helps direct clinical management. Clinical Relevance Patients diagnosed with ROP may have a mutation in the FZD4 gene and display characteristics consistent with FEVR. Analysis of the FZD4 gene should be considered. PMID- 20008723 TI - Swan M. Burnett, MD, PhD: the forgotten father of Little Lord Fauntleroy. PMID- 20008722 TI - Adverse events after pars plana vitrectomy among medicare beneficiaries. AB - OBJECTIVES: To assess the complication rates of pars plana vitrectomy (PPV) among older Americans and to determine whether rates of adverse events and additional operations have changed during the past decade. METHODS: Claims data were reviewed to identify all adults aged 68 years or older in the 5% Medicare sample who underwent their first PPV during 1994-1995, 1999-2000, and 2004-2005. One year rates of severe complications (endophthalmitis, suprachoroidal hemorrhage, or retinal detachment), less severe complications, receipt of an additional operation, and blindness were calculated and compared among the 3 groups using Cox regression. Analyses were adjusted for prior adverse events (during the previous 3 years), demographic characteristics, and comorbid conditions. RESULTS: The 1994-1995, 1999-2000, and 2004-2005 cohorts had 3263, 5064, and 5263 patients, respectively. The 1-year severe complication rates did not differ among the 3 groups (range, 4.8%-5.5%). The hazard of a less severe complication or an additional operation was higher in the 2004-2005 cohort than in the earlier cohorts (P < .05 for all comparisons). The hazard of endophthalmitis was higher in black individuals (P = .07) and those of other races (P = .02) than in white patients. CONCLUSIONS: During the past decade, rates of severe complications after PPV remained stable, but rates of less severe complications and subsequent operations increased. Future studies should explore the potential factors that explain these changes and the alarming elevated incidence of post-PPV endophthalmitis among nonwhite individuals. PMID- 20008724 TI - Intravitreal corticosteroids for diabetic retinopathy prevention. PMID- 20008725 TI - Bringing better treatments to patients faster by expanding access. PMID- 20008726 TI - The 100 most influential journals. PMID- 20008727 TI - Summary results and recommendations from the age-related eye disease study. PMID- 20008728 TI - Legacy of the age-related eye disease [corrected] study. PMID- 20008729 TI - Liquid nitrogen cryotherapy of conjunctival lymphangiectasia: a case series. PMID- 20008730 TI - Macular hole closure and visual improvement with topical nonsteroidal treatment. PMID- 20008731 TI - Ocular vaccinia with severe restriction of extraocular motility. PMID- 20008732 TI - Bitemporal hemianopia caused by retinal disease. PMID- 20008736 TI - Tuberculous pseudoaneurysms of the thoracic aorta: comprehensive evaluation by cardiovascular magnetic resonance. PMID- 20008733 TI - Slit2 inhibits glioma cell invasion in the brain by suppression of Cdc42 activity. AB - Acquisition of insidious invasiveness by malignant glioma cells involves multiple genetic alterations in signaling pathways. Slit2, a chemorepulsive factor, controls cell migration of neuronal and glial cells during development and inhibits chemotaxic migration of various types of cells in vitro. However, the role of Slit2 in vitro remains controversial, and the biological significance of Slit2 expression in cancer cell invasion in vivo has not yet been determined. In the present study, we characterized the effects of Slit2 expression on the migration and invasion of invasive glioma cells in vitro and in vivo. By reverse transcriptase polymerase chain reaction (PCR) analyses, Slit2 was found to be expressed at lower levels in primary glioma specimens and invasive glioma cells compared with normal human brain cells and astrocytes. Ectopic expression of Slit2 or treatment with recombinant Slit2 on glioma cells attenuates cell migration and invasion through inhibition of Cdc42 activity in vitro. Cellular depletion of Robo1, a cognate receptor for Slit2, prevented Slit2 inhibition of Cdc42 activity and glioma cell migration. In vivo, expression of Slit2 by invasive SNB19 glioma cells markedly inhibited glioma cell infiltration into the brain of mice. Moreover, impediment of glioma cell invasion by Slit2 did not affect the expression of N-cadherin and beta-catenin in glioma cells. These results provide the first evidence demonstrating that Slit2-Robo1 inhibits glioma invasion through attenuating Cdc42 activity in vitro and in the brain. Understanding the mechanisms of Slit2-Robo1 inhibition of glioma cell invasion will foster new treatments for malignant gliomas. PMID- 20008737 TI - Letter by Spyropoulos regarding article, "Physician alerts to prevent symptomatic venous thromboembolism in hospitalized patients". PMID- 20008739 TI - Inhibitors of platelet adhesion. PMID- 20008741 TI - Cardiac herniation after minimally invasive cardiac surgery: an unusual potentially lethal complication. PMID- 20008740 TI - Electrophysiological challenges of cell-based myocardial repair. PMID- 20008742 TI - Systematic review: comparative effectiveness of core-needle and open surgical biopsy to diagnose breast lesions. AB - BACKGROUND: Most women undergoing breast biopsy are found not to have cancer. PURPOSE: To compare the accuracy and harms of different breast biopsy methods in average-risk women suspected of having breast cancer. DATA SOURCES: Databases, including MEDLINE and EMBASE, searched from 1990 to September 2009. STUDY SELECTION: Studies that compared core-needle biopsy diagnoses with open surgical diagnoses or clinical follow-up. DATA EXTRACTION: Data were abstracted by 1 of 3 researchers and verified by the primary investigator. DATA SYNTHESIS: 33 studies of stereotactic automated gun biopsy; 22 studies of stereotactic-guided, vacuum assisted biopsy; 16 studies of ultrasonography-guided, automated gun biopsy; 7 studies of ultrasonography-guided, vacuum-assisted biopsy; and 5 studies of freehand automated gun biopsy met the inclusion criteria. Low-strength evidence showed that core-needle biopsies conducted under stereotactic guidance with vacuum assistance distinguished between malignant and benign lesions with an accuracy similar to that of open surgical biopsy. Ultrasonography-guided biopsies were also very accurate. The risk for severe complications is lower with core needle biopsy than with open surgical procedures (<1% vs. 2% to 10%). Moderate strength evidence showed that women in whom breast cancer was initially diagnosed by core-needle biopsy were more likely than women with cancer initially diagnosed by open surgical biopsy to be treated with a single surgical procedure (random effects odds ratio, 13.7 [95% CI, 5.5 to 34.6]). LIMITATION: The strength of evidence was rated low for accuracy outcomes because the studies did not report important details required to assess the risk for bias. CONCLUSION: Stereotactic- and ultrasonography-guided core-needle biopsy procedures seem to be almost as accurate as open surgical biopsy, with lower complication rates. PRIMARY FUNDING SOURCE: Agency for Healthcare Research and Quality. PMID- 20008743 TI - Teaching primary care in community health centers: addressing the workforce crisis for the underserved. AB - Universal coverage and multiple initiatives to improve health care delivery are crucial components of health care reform. However, the missing link has been a plan to rapidly address the primary care workforce crisis for the underserved. The authors propose a link between primary care graduate medical education and care for the underserved in community health centers, where expansion will be necessary for the anticipated increase in Medicaid and insured patients. This can be achieved by establishing primary care teaching health centers in expanded community health centers, which have established a patient-centered medical home practice environment. Residents would receive their final year of training in these centers, and then have the incentive of National Health Service Corps debt repayment if they subsequently practice in an underserved area. Primary care residents being trained in this setting would immediately increase the clinical capacity of community health centers and ultimately expand the primary care physician workforce. This proposal addresses the primary care physician workforce crisis and the associated key problems of limited access for the underserved and suboptimal primary care graduate medical education. PMID- 20008744 TI - Summaries for patients. Assessing the best way to prevent spread of influenza. PMID- 20008745 TI - Summaries for patients. Chronic health problems seem to influence the relationship of blood sugar control and cardiovascular events in patients with type 2 diabetes. PMID- 20008746 TI - ACP Journal Club. PET plus CT was better than conventional methods for correctly upstaging early NSCLC. PMID- 20008747 TI - ACP Journal Club. Screening with spiral CT did not reduce lung cancer mortality in older male smokers. PMID- 20008748 TI - ACP Journal Club. Sensitive cardiac troponin assays were more accurate than a standard troponin assay for early diagnosis of AMI. PMID- 20008749 TI - ACP Journal Club. Anemia was independently associated with mortality in very old persons. PMID- 20008750 TI - ACP Journal Club. Continuous androgen-deprivation therapy increased risk for diabetes and fragility fractures in older men with prostate cancer. PMID- 20008752 TI - ACP Journal Club. Editorial: new and underutilized features of ACP Journal Club PLUS and ACPJC.org: stellar articles, searches, and succinct synopses of the principles and practice of "personalized" evidence-based medicine. PMID- 20008751 TI - ACP Journal Club. SSRIs in early pregnancy were associated with increased risk for septal heart defects but not major congenital malformations overall. PMID- 20008753 TI - ACP Journal Club. Ticagrelor was more effective than clopidogrel, with no increase in major bleeding in acute coronary syndromes. PMID- 20008754 TI - ACP Journal Club. A Mediterranean-style diet reduced need for glucose-lowering drugs more than a low-fat diet in type 2 diabetes. PMID- 20008755 TI - ACP Journal Club. Human papillomavirus 16/18 AS04-adjuvanted vaccine prevented cervical intraepithelial neoplasia in young women. PMID- 20008756 TI - ACP Journal Club. Review: pregabalin reduces chronic neuropathic pain in adults. PMID- 20008757 TI - ACP Journal Club. Vertebroplasty was not effective for painful osteoporotic vertebral fractures. PMID- 20008758 TI - ACP Journal Club. Vertebroplasty was not effective for painful osteoporotic vertebral fractures. PMID- 20008759 TI - Effectiveness and cost-effectiveness of vaccination against pandemic influenza (H1N1) 2009. AB - BACKGROUND: Decisions on the timing and extent of vaccination against pandemic (H1N1) 2009 virus are complex. OBJECTIVE: To estimate the effectiveness and cost effectiveness of pandemic influenza (H1N1) vaccination under different scenarios in October or November 2009. DESIGN: Compartmental epidemic model in conjunction with a Markov model of disease progression. DATA SOURCES: Literature and expert opinion. TARGET POPULATION: Residents of a major U.S. metropolitan city with a population of 8.3 million. TIME HORIZON: Lifetime. PERSPECTIVE: Societal. INTERVENTIONS: Vaccination in mid-October or mid-November 2009. OUTCOME MEASURES: Infections and deaths averted, costs, quality-adjusted life-years (QALYs), and incremental cost-effectiveness. RESULTS OF BASE-CASE ANALYSIS: Assuming each primary infection causes 1.5 secondary infections, vaccinating 40% of the population in October or November would be cost-saving. Vaccination in October would avert 2051 deaths, gain 69 679 QALYs, and save $469 million compared with no vaccination; vaccination in November would avert 1468 deaths, gain 49 422 QALYs, and save $302 million. RESULTS OF SENSITIVITY ANALYSIS: Vaccination is even more cost-saving if longer incubation periods, lower rates of infectiousness, or increased implementation of nonpharmaceutical interventions delay time to the peak of the pandemic. Vaccination saves fewer lives and is less cost-effective if the epidemic peaks earlier than mid-October. LIMITATIONS: The model assumed homogenous mixing of case-patients and contacts; heterogeneous mixing would result in faster initial spread, followed by slower spread. Additional costs and savings not included in the model would make vaccination more cost-saving. CONCLUSION: Earlier vaccination against pandemic (H1N1) 2009 prevents more deaths and is more cost-saving. Complete population coverage is not necessary to reduce the viral reproductive rate sufficiently to help shorten the pandemic. PRIMARY FUNDING SOURCE: Agency for Healthcare Research and Quality and National Institute on Drug Abuse. PMID- 20008761 TI - Comorbidity affects the relationship between glycemic control and cardiovascular outcomes in diabetes: a cohort study. AB - BACKGROUND: Recent studies have shown mixed results regarding the effectiveness of intensive glucose-lowering therapy in reducing risk for cardiovascular events. OBJECTIVE: To determine whether attaining hemoglobin A(1c) (HbA(1c)) targets of 6.5% or less or 7.0% or less for glycemic control at baseline provides differential benefits for patients with high versus low-to-moderate levels of comorbidity. DESIGN: 5-year longitudinal observational study of patients with type 2 diabetes. Patients were categorized into high and low-to-moderate comorbidity subgroups by using the Total Illness Burden Index (TIBI), a validated patient-reported measure of comorbidity. SETTING: 101 diabetes outpatient clinics and 103 general practitioners' clinics in Italy. PATIENTS: 2613 (83%) of 3074 patients with type 2 diabetes, sampled randomly from diabetes outpatient clinic rosters and recruited consecutively from general practitioners' clinics, who completed the baseline questionnaire. MEASUREMENTS: TIBI score, total mortality, and incident cardiovascular events. Hazard ratios (HRs) were adjusted for age and sex. RESULTS: Attaining an HbA(1c) level of 6.5% or less at baseline was associated with lower 5-year incidence of cardiovascular events in the low-to moderate comorbidity subgroup (adjusted HR, 0.60 [95% CI, 0.42 to 0.85]; P = 0.005) but not in the high comorbidity subgroup (adjusted HR, 0.92 [CI, 0.68 to 1.25]; P = 0.61; P for subgroup by HbA(1c) interaction = 0.048). Similarly, attaining a baseline HbA(1c) level of 7.0% predicted fewer cardiovascular events in the low-to-moderate comorbidity subgroup (adjusted HR, 0.61 (CI, 0.44 to 0.83; P = 0.001) but not in the high comorbidity subgroup (adjusted HR, 0.88 [CI, 0.66 to 1.17]; P = 0.38; P for subgroup by HbA(1c) interaction = 0.093). LIMITATIONS: The observational nature of the study does not allow causal inference. The length of the data collection period was limited. Information on clinical management was not available. CONCLUSION: Patients with the high levels of comorbidity common in type 2 diabetes may receive diminished cardiovascular benefit from intensive blood glucose control. Comorbidity should be considered when tailoring glucose lowering therapy in patients with type 2 diabetes. PRIMARY FUNDING SOURCE: Pfizer of Italy. PMID- 20008760 TI - Effectiveness and cost-effectiveness of expanded antiviral prophylaxis and adjuvanted vaccination strategies for an influenza A (H5N1) pandemic. AB - BACKGROUND: The pandemic potential of influenza A (H5N1) virus is a prominent public health concern of the 21st century. OBJECTIVE: To estimate the effectiveness and cost-effectiveness of alternative pandemic (H5N1) mitigation and response strategies. DESIGN: Compartmental epidemic model in conjunction with a Markov model of disease progression. DATA SOURCES: Literature and expert opinion. TARGET POPULATION: Residents of a U.S. metropolitan city with a population of 8.3 million. TIME HORIZON: Lifetime. PERSPECTIVE: Societal. INTERVENTIONS: 3 scenarios: 1) vaccination and antiviral pharmacotherapy in quantities similar to those currently available in the U.S. stockpile (stockpiled strategy), 2) stockpiled strategy but with expanded distribution of antiviral agents (expanded prophylaxis strategy), and 3) stockpiled strategy but with adjuvanted vaccine (expanded vaccination strategy). All scenarios assumed standard nonpharmaceutical interventions. OUTCOME MEASURES: Infections and deaths averted, costs, quality-adjusted life-years (QALYs), and incremental cost effectiveness. RESULTS OF BASE-CASE ANALYSIS: Expanded vaccination was the most effective and cost-effective of the 3 strategies, averting 68% of infections and deaths and gaining 404 030 QALYs at $10 844 per QALY gained relative to the stockpiled strategy. RESULTS OF SENSITIVITY ANALYSIS: Expanded vaccination remained incrementally cost-effective over a wide range of assumptions. LIMITATIONS: The model assumed homogenous mixing of cases and contacts; heterogeneous mixing would result in faster initial spread, followed by slower spread. We did not model interventions for children or older adults; the model is not designed to target interventions to specific groups. CONCLUSION: Expanded adjuvanted vaccination is an effective and cost-effective mitigation strategy for an influenza A (H5N1) pandemic. Expanded antiviral prophylaxis can help delay the pandemic while additional strategies are implemented. PRIMARY FUNDING SOURCE: National Institutes of Health and Agency for Healthcare Research and Quality. PMID- 20008762 TI - Systematic review: comparative effectiveness of angiotensin-converting enzyme inhibitors or angiotensin II-receptor blockers for ischemic heart disease. AB - BACKGROUND: Patients with ischemic heart disease and preserved ventricular function experience considerable morbidity and mortality despite standard medical therapy. PURPOSE: To compare benefits and harms of using angiotensin-converting enzyme (ACE) inhibitors, angiotensin II-receptor blockers (ARBs), or combination therapy in adults with stable ischemic heart disease and preserved ventricular function. DATA SOURCES: MEDLINE, EMBASE, Cochrane Central Register of Controlled Trials, and Cochrane Database of Systematic Reviews (earliest date, July 2009) were searched without language restrictions. STUDY SELECTION: Two independent investigators screened citations for trials of at least 6 months' duration that compared ACE inhibitors, ARBs, or combination therapy with placebo or active control and reported any of several clinical outcomes. DATA EXTRACTION: Using standardized protocols, 2 independent investigators extracted information about study characteristics and rated the quality and strength of evidence. Disagreement was resolved by consensus. DATA SYNTHESIS: 41 studies met eligibility criteria. Moderate- to high-strength evidence (7 trials; 32 559 participants) showed that ACE inhibitors reduce the relative risk (RR) for total mortality (RR, 0.87 [95% CI, 0.81 to 0.94]) and nonfatal myocardial infarction (RR, 0.83 [CI, 0.73 to 0.94]) but increase the RR for syncope (RR, 1.24 [CI, 1.02 to 1.52]) and cough (RR, 1.67 [CI, 1.22 to 2.29]) compared with placebo. Low strength evidence (1 trial; 5926 participants) suggested that ARBs reduce the RR for the composite end point of cardiovascular mortality, nonfatal myocardial infarction, or stroke (RR, 0.88 [CI, 0.77 to 1.00]) but not for the individual components. Moderate-strength evidence (1 trial; 25 620 participants) showed similar effects on total mortality (RR, 1.07 [CI, 0.98 to 1.16]) and myocardial infarction (RR, 1.08 [CI, 0.94 to 1.23]) but an increased risk for discontinuations because of hypotension (P < 0.001) and syncope (P = 0.035) with combination therapy compared with ACE inhibitors alone. LIMITATIONS: Many studies either did not assess or did not report harms in a systematic manner. Many studies did not adequately report benefits or harms by various patient subgroups. CONCLUSION: Adding an ACE inhibitor to standard medical therapy improves outcomes, including reduced risk for mortality and myocardial infarctions, in some patients with stable ischemic heart disease and preserved ventricular function. Less evidence supports a benefit of ARB therapy, and combination therapy seems no better than ACE inhibitor therapy alone and increases harms. PRIMARY FUNDING SOURCE: Agency for Healthcare Research and Quality. PMID- 20008763 TI - The modern crystal ball: influenza forecasting with mathematical models. PMID- 20008764 TI - The role of glycemia management in the prevention of cardiovascular disease- starting over? PMID- 20008765 TI - Three degrees of separation. PMID- 20008766 TI - The CKD-EPI equation and MDRD study equation find similar prevalence of chronic kidney disease in Asian populations. PMID- 20008767 TI - Did Akhenaten have the Antley-Bixler syndrome? PMID- 20008768 TI - Relevance of molecular markers in prostate cancer. PMID- 20008769 TI - A pain in the neck: probiotics for ulcerative colitis. PMID- 20008771 TI - Retraction. Combination treatment of angiotensin-II receptor blocker and angiotensin-converting-enzyme inhibitor in non-diabetic renal disease (COOPERATE): a randomised controlled trial. PMID- 20008770 TI - Challenges of interpreting results from analyses that use extensive claims data. PMID- 20008772 TI - Inhibitor resistance in the KPC-2 beta-lactamase, a preeminent property of this class A beta-lactamase. AB - As resistance determinants, KPC beta-lactamases demonstrate a wide substrate spectrum that includes carbapenems, oxyimino-cephalosporins, and cephamycins. In addition, clinical strains harboring KPC-type beta-lactamases are often identified as resistant to standard beta-lactam-beta-lactamase inhibitor combinations in susceptibility testing. The KPC-2 carbapenemase presents a significant clinical challenge, as the mechanistic bases for KPC-2-associated phenotypes remain elusive. Here, we demonstrate resistance by KPC-2 to beta lactamase inhibitors by determining that clavulanic acid, sulbactam, and tazobactam are hydrolyzed by KPC-2 with partition ratios (kcat/kinact ratios, where kinact is the rate constant of enzyme inactivation) of 2,500, 1,000, and 500, respectively. Methylidene penems that contain an sp2-hybridized C3 carboxylate and a bicyclic R1 side chain (dihydropyrazolo[1,5-c][1,3]thiazole [penem 1] and dihydropyrazolo[5,1-c][1,4]thiazine [penem 2]) are potent inhibitors: Km of penem 1, 0.06+/-0.01 microM, and Km of penem 2, 0.006+/-0.001 microM. We also demonstrate that penems 1 and 2 are mechanism-based inactivators, having partition ratios (kcat/kinact ratios) of 250 and 50, respectively. To understand the mechanism of inhibition by these penems, we generated molecular representations of both inhibitors in the active site of KPC-2. These models (i) suggest that penem 1 and penem 2 interact differently with active site residues, with the carbonyl of penem 2 being positioned outside the oxyanion hole and in a less favorable position for hydrolysis than that of penem 1, and (ii) support the kinetic observations that penem 2 is the better inhibitor (kinact/Km=6.5+/-0.6 microM(-1) s(-1)). We conclude that KPC-2 is unique among class A beta-lactamases in being able to readily hydrolyze clavulanic acid, sulbactam, and tazobactam. In contrast, penem-type beta-lactamase inhibitors, by exhibiting unique active site chemistry, may serve as an important scaffold for future development and offer an attractive alternative to our current beta-lactamase inhibitors. PMID- 20008773 TI - Murine model of a disseminated infection by the novel fungus Fonsecaea monophora and successful treatment with posaconazole. AB - We have evaluated the efficacy of posaconazole, amphotericin B, and itraconazole in a murine model of disseminated infection by Fonsecaea monophora. Of these three antifungal drugs tested, posaconazole prolonged survival significantly and reduced the fungal load in most of the organs tested. Bioassay studies demonstrated the relationship between posaconazole levels and dose escalation in serum and brain tissue. Posaconazole may have a clinical role in the treatment of disseminated infections by F. monophora. PMID- 20008775 TI - Newly identified antibacterial compounds are topoisomerase poisons in African trypanosomes. AB - Human African trypanosomiasis, caused by the Trypanosoma brucei protozoan parasite, is fatal when left untreated. Current therapies are antiquated, and there is a need for new pharmacologic agents against T. brucei targets that have no human ortholog. Trypanosomes have a single mitochondrion with a unique mitochondrial DNA, known as kinetoplast DNA (kDNA), a topologically complex network that contains thousands of interlocking circular DNAs, termed minicircles (approximately 1 kb) and maxicircles (approximately 23 kb). Replication of kDNA depends on topoisomerases, enzymes that catalyze reactions that change DNA topology. T. brucei has an unusual type IA topoisomerase that is dedicated to kDNA metabolism. This enzyme has no ortholog in humans, and RNA interference (RNAi) studies have shown that it is essential for parasite survival, making it an ideal drug target. In a large chemical library screen, two compounds were recently identified as poisons of bacterial topoisomerase IA. We found that these compounds are trypanocidal in the low micromolar range and that they promote the formation of linearized minicircles covalently bound to protein on the 5' end, consistent with the poisoning of mitochondrial topoisomerase IA. Surprisingly, however, band depletion studies showed that it is topoisomerase IImt, and not topoisomerase IAmt, that is trapped. Both compounds are planar aromatic polycyclic structures that intercalate into and unwind DNA. These findings reinforce the utility of topoisomerase IImt as a target for development of new drugs for African sleeping sickness. PMID- 20008774 TI - Glycerol monolaurate inhibits Candida and Gardnerella vaginalis in vitro and in vivo but not Lactobacillus. AB - We investigated the effects of glycerol monolaurate (GML) on Lactobacillus, Candida, and Gardnerella vaginalis human vaginal microflora. Our previous work demonstrated that 6 months of GML treatment vaginally does not alter lactobacillus counts in monkeys. Candida and G. vaginalis are commonly associated with vaginal infections in women, many becoming chronic or recurrent. In vitro growth inhibition studies determined the effects of GML (0 to 500 microg/ml) against multiple Candida species and G. vaginalis. A randomized, double-blind study investigated the effects of GML on vaginal microflora Lactobacillus, Candida, and G. vaginalis in colonized or infected women (n=36). Women self administered intravaginal gels containing 0% (n=14), 0.5% (n=13), or 5% (n=9) GML every 12 h for 2 days. Vaginal swabs were collected before and immediately after the first gel administration and 12 h after the final gel administration. Swabs were tested for Lactobacillus, Candida, G. vaginalis, and GML. In vitro GML concentrations of 500 microg/ml were candicidal for all species tested, while a concentration of 10 microg/ml was bactericidal for G. vaginalis. Control and GML gels applied vaginally in women did not alter vaginal pH or Lactobacillus counts. Control gels reduced G. vaginalis counts but not Candida counts, whereas GML gels reduced both Candida and G. vaginalis. No adverse events were reported by participating women. GML is antimicrobial for Candida and G. vaginalis in vitro. Vaginal GML gels in women do not affect Lactobacillus negatively but significantly reduce Candida and G. vaginalis. PMID- 20008776 TI - Constitutive SoxS expression in a fluoroquinolone-resistant strain with a truncated SoxR protein and identification of a new member of the marA-soxS-rob regulon, mdtG. AB - Elevated levels of fluoroquinolone resistance are frequently found among Escherichia coli clinical isolates. This study investigated the antibiotic resistance mechanisms of strain NorE5, derived in vitro by exposing an E. coli clinical isolate, PS5, to two selection steps with increasing concentrations of norfloxacin. In addition to the amino acid substitution in GyrA (S83L) present in PS5, NorE5 has an amino acid change in ParC (S80R). Furthermore, we now find by Western blotting that NorE5 has a multidrug resistance phenotype resulting from the overexpression of the antibiotic resistance efflux pump AcrAB-TolC. Microarray and gene fusion analyses revealed significantly increased expression in NorE5 of soxS, a transcriptional activator of acrAB and tolC. The high soxS activity is attributable to a frameshift mutation that truncates SoxR, rendering it a constitutive transcriptional activator of soxS. Furthermore, microarray and reverse transcription-PCR analyses showed that mdtG (yceE), encoding a putative efflux pump, is overexpressed in the resistant strain. SoxS, MarA, and Rob activated an mdtG::lacZ fusion, and SoxS was shown to bind to the mdtG promoter, showing that mdtG is a member of the marA-soxS-rob regulon. The mdtG marbox sequence is in the backward or class I orientation within the promoter, and its disruption resulted in a loss of inducibility by MarA, SoxS, and Rob. Thus, chromosomal mutations in parC and soxR are responsible for the increased antibiotic resistance of NorE5. PMID- 20008777 TI - aac(6')-Ib-cr genotyping by simultaneous high-resolution melting analyses of an unlabeled probe and full-length amplicon. AB - We have developed a time- and cost-efficient one-step closed-tube assay for genotyping of aac(6')-Ib-cr that is capable of distinguishing between the two genetic aac(6')-Ib-cr variants. Our genotyping assay uses the combined information of simultaneously acquired high-resolution melting data from an unlabeled probe and the full-length amplicon. PMID- 20008778 TI - Selection of mutations to detect multidrug-resistant Mycobacterium tuberculosis strains in Shanghai, China. AB - Novel tools are urgently needed for the rapid, reliable detection of multidrug resistant (MDR) and extensively drug-resistant (XDR) strains of Mycobacterium tuberculosis. To develop such tools, we need information about the frequency and distribution of the mycobacterial mutations and genotypes that are associated with phenotypic drug resistance. In a population-based study, we sequenced specific genes of M. tuberculosis that were associated with resistance to rifampin and isoniazid in 242 phenotypically MDR isolates and 50 phenotypically pan-susceptible isolates from tuberculosis (TB) cases in Shanghai, China. We estimated the sensitivity and specificity of the mutations, using the results of conventional, culture-based phenotypic drug susceptibility testing as the standard. We detected mutations within the 81-bp core region of rpoB in 96.3% of phenotypically MDR isolates. Mutations in two structural genes (katG and inhA) and two regulatory regions (the promoter of mabA-inhA and the intergenic region of oxyR-ahpC) were found in 89.3% of the MDR isolates. In total, 88.0% (213/242 strains) of the phenotypic MDR strains were confirmed by mutations in the sequenced regions. Mutations in embB306 were also considered a marker for MDR and significantly increased the sensitivity of the approach. Based on our findings, an approach that prospectively screens for mutations in 11 sites of the M. tuberculosis genome (rpoB531, rpoB526, rpoB516, rpoB533, and rpoB513, katG315, inhA-15, ahpC-10, ahpC-6, and ahpC-12, and embB306) could detect 86.8% of MDR strains in Shanghai. This study lays the foundation for the development of a rapid, reliable molecular genetic test to detect MDR strains of M. tuberculosis in China. PMID- 20008780 TI - Identification of a plasmid-borne resistance gene cluster comprising the resistance genes erm(T), dfrK, and tet(L) in a porcine methicillin-resistant Staphylococcus aureus ST398 strain. AB - A novel plasmid-borne resistance gene cluster comprising the genes erm(T) for macrolide-lincosamide-streptogramin B resistance, dfrK for trimethoprim resistance, and tet(L) for tetracycline resistance was identified in a porcine methicillin-resistant Staphylococcus aureus sequence type 398 (ST398) strain. This erm(T)-dfrK-tet(L) region was flanked by copies of the novel IS element ISSau10. The erm(T) region resembled that of Streptococcus pyogenes plasmid pRW35. The erm(T) gene of pKKS25 was expressed constitutively due to a 57-bp deletion in the erm(T) translational attenuator. PMID- 20008779 TI - Resistance-associated mutations to etravirine (TMC-125) in antiretroviral-naive patients infected with non-B HIV-1 subtypes. AB - Susceptibility to etravirine (ETR), an expanded-spectrum nonnucleoside reverse transcriptase inhibitor (NNRTI), is dependent on the type and number of NNRTI resistance-associated mutations (RAMs). Studies have shown that some HIV-1 subtypes may have natural polymorphisms described as ETR RAMs. This study addresses the prevalence of ETR RAMs in treatment-naive patients infected with HIV-1 non-B subtypes and its potential impact on ETR susceptibility. The prevalence of ETR RAMs in 726 antiretroviral-naive patients infected with non-B HIV-1 subtypes was studied. ETR genotypic resistance was interpreted according to Agence Nationale de Recherches sur le SIDA and Stanford algorithms. NNRTI phenotypic susceptibilities of samples with at least one ETR RAM were measured. Overall, 75 (10.3%) of 726 sequences harbored at least one ETR RAM: sequences from 72 patients (10%) each had one ETR RAM, and sequences from 3 patients (0.4%) each had two ETR RAMs (V90I and Y181C in one case and V90I and A98G in two cases). None of the viruses had three or more ETR RAMs, and none were consequently classified as resistant to ETR. All sequences with two ETR RAMs belonged to subtype CRF02_AG. The presence of one ETR RAM was statistically more frequent in subtype CRF02_AG than in other non-B subtypes (P=0.004). Three new mutation profiles (E138A and V179I, Y181C and H221Y, and V90I and Y181C) showing decreased ETR phenotypic susceptibility were identified. In conclusion, although the prevalence of ETR RAMs in treatment-naive patients infected with non-B HIV-1 subtypes was 10%, in most cases this had no significant impact on ETR susceptibility. However, the transmission of drug-resistant viruses with Y181C in a non-B genetic background has a potential for impact on ETR susceptibility. PMID- 20008781 TI - Comparative study of the mutant prevention concentrations of moxifloxacin, levofloxacin, and gemifloxacin against pneumococci. AB - We tested the propensity of three quinolones to select for resistant Streptococcus pneumoniae mutants by determining the mutant prevention concentration (MPC) against 100 clinical strains, some of which harbored mutations in type II topoisomerases. Compared with levofloxacin and gemifloxacin, moxifloxacin had the lowest number of strains with MPCs above the susceptibility breakpoint (P<0.001), thus representing a lower selective pressure for proliferation of resistant mutants. Only moxifloxacin gave a 50% MPC (MPC50) value (1 microg/ml) within the susceptible range. PMID- 20008782 TI - In vitro resistance selection studies of RLP068/Cl, a new Zn(II) phthalocyanine suitable for antimicrobial photodynamic therapy. AB - Resistance to antimicrobial agents is emerging in a wide variety of nosocomial and community-acquired pathogens. The development of alternative therapies against nosocomial infections caused by clinically relevant pathogens represents a major public health concern. RLP068/Cl is a novel Zn(II) phthalocyanine proposed as a photosensitizer suitable for antimicrobial photodynamic therapy (APDT) for localized infections. Its ability, following activation by light, to induce resistance in three major human pathogens after 20 daily passages was studied. Simultaneously for the same strains, the ability of daily sequential subcultures in subinhibitory concentrations of RLP068/Cl to develop resistant mutants without illumination was evaluated. We demonstrate that 20 consecutive APDT treatments with RLP068/Cl did not result in any resistant mutants and that, in dark conditions, only Staphylococcus aureus strains had increased MICs of RLP068/Cl. However, even in this case, the susceptibility of the mutated bacteria to APDT was not affected by their MIC increase. PMID- 20008783 TI - Characterization of small ColE-like plasmids mediating widespread dissemination of the qnrB19 gene in commensal enterobacteria. AB - In this work, we have characterized two small ColE-like plasmids (pECY6-7, 2.7 kb in size, and pECC14-9, of 3.0 kb), encoding the QnrB19 quinolone resistance determinant, that were carried by several clonally unrelated quinolone-resistant commensal Escherichia coli strains isolated from healthy children living in different urban areas of Peru and Bolivia. The two plasmids are closely related to each other and carry the qnrB19 gene as the sole resistance determinant, located in a conserved genetic context between the plasmid RNAII sequence (which controls plasmid replication) and the plasmid Xer site (involved in plasmid dimer resolution). ISEcp1-like or other putative insertion sequences are not present in the qnrB19-flanking regions or elsewhere on the plasmids. Since we previously observed a high prevalence (54%) of qnrB genes in the metagenomes of commensal enterobacteria from the same population of healthy children, the presence of pECY6-7- and pECC14-9-like plasmids in those qnrB-positive metagenomes was investigated by PCR mapping. Both plasmids were found to be highly prevalent (67% and 16%, respectively) in the qnrB-positive metagenomes, suggesting that dissemination of these small plasmids played a major role in the widespread dissemination of qnrB genes observed in commensal enterobacteria from healthy children living in those areas. PMID- 20008785 TI - A dynamic prognostic model to predict survival in primary myelofibrosis: a study by the IWG-MRT (International Working Group for Myeloproliferative Neoplasms Research and Treatment). AB - Age older than 65 years, hemoglobin level lower than 100 g/L (10 g/dL), white blood cell count greater than 25 x 10(9)/L, peripheral blood blasts 1% or higher, and constitutional symptoms have been shown to predict poor survival in primary myelofibrosis (PMF) at diagnosis. To investigate whether the acquisition of these factors during follow-up predicts survival, we studied 525 PMF patients regularly followed. All 5 variables had a significant impact on survival when analyzed as time-dependent covariates in a multivariate Cox proportional hazard model and were included in 2 separate models, 1 for all patients (Dynamic International Prognostic Scoring System [DIPSS]) and 1 for patients younger than 65 years (age adjusted DIPSS). Risk factors were assigned score values based on hazard ratios (HRs). Risk categories were low, intermediate-1, intermediate-2, and high in both models. Survival was estimated by the HR. When shifting to the next risk category, the HR was 4.13 for low risk, 4.61 for intermediate-1, and 2.54 for intermediate-2 according to DIPSS; 3.97 for low risk, 2.84 for intermediate-1, and 1.81 for intermediate-2 according to the age-adjusted DIPSS. The novelty of these models is the prognostic assessment of patients with PMF anytime during their clinical course, which may be useful for treatment decision-making. PMID- 20008784 TI - Evaluation of WLBU2 peptide and 3-O-octyl-sn-glycerol lipid as active ingredients for a topical microbicide formulation targeting Chlamydia trachomatis. AB - Topical microbicides for prevention of sexually transmitted diseases (STDs) would be especially useful for women who are not able to persuade their partner(s) to take precautions. Many topical microbicides are in various stages of development, based on a variety of active ingredients. We investigated the in vitro activity of an engineered antimicrobial peptide (WLBU2) and a lipid (3-O-octyl-sn-glycerol [3-OG]) which could potentially be used as active ingredients in such a product. Using commercially available cytotoxicity reagents [Alamar Blue, 3-(4,5-dimethyl 2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide (MTT), and lactate dehydrogenase (LDH)], we first determined the toxicity of WLBU2 and 3-OG to the host cells in our assay procedure and excluded toxic concentrations from further testing. To determine activity against Chlamydia trachomatis, we used an assay previously developed by our laboratory in which chlamydial elementary bodies (EBs) were exposed to microbicides prior to contact with epithelial cells: the minimum (microbi)cidal concentration (MCC) assay. To further simulate conditions of transmission, we carried out the same assay in the presence of a simulated vaginal fluid, a simulated seminal fluid, human serum albumin, and a range of pH values which might be found in the human vagina at the time of exposure. Last, we tested WLBU2 and 3-OG in combination to determine if adding them together resulted in synergistic activity. We found that WLBU2 and 3-OG both have excellent activity in vitro against C. trachomatis and significantly more activity when added together. The simulated fluids reduced activity, but the synergy seen is good evidence that they would be effective when combined in a microbicide formulation. PMID- 20008786 TI - Erythropoietin accelerates smooth muscle cell-rich vascular lesion formation in mice through endothelial cell activation involving enhanced PDGF-BB release. AB - In this study, the effect of human erythropoietin Delta (Epo) on smooth muscle cell (SMC)-rich lesions was evaluated. Mice, of which the left carotid artery was ligated, were treated with suberythropoietic as well as erythropoietic doses of Epo and both doses of Epo enhanced SMC-rich lesion formation. No association was observed between hemoglobin levels and lesion size. Moreover, endothelial progenitor cell (EPC) numbers in the peripheral blood increased only in the erythropoietic dosing group, indicating that EPC numbers did not correlate with lesion size. Immunohistochemical analysis revealed that Epo-mediated enhancement of lesion formation correlates with increased signal transducer and activator of transcription 5 (Stat5) phosphorylation in the vessel wall. Experiments performed in cultured vascular cells demonstrated that Epo robustly induced phosphorylation of Stat5 in human umbilical vein endothelial cells (HUVECs), but only very weakly in SMCs. In tumor necrosis factor-alpha (TNFalpha)-activated HUVECS, Epo induced expression of platelet-derived growth factor B (PDGF-B), which was at least partially responsible for the induction of Stat5 phosphorylation in SMCs by HUVEC conditioned medium. In conclusion, in mice Epo accelerates SMC-rich neointima formation, which correlates with increased Stat5 phosphorylation in the vessel wall but is independent of erythrocyte and EPC numbers. PMID- 20008787 TI - Constitutively active AKT depletes hematopoietic stem cells and induces leukemia in mice. AB - Human cancers, including acute myeloid leukemia (AML), commonly display constitutive phosphoinositide 3-kinase (PI3K) AKT signaling. However, the exact role of AKT activation in leukemia and its effects on hematopoietic stem cells (HSCs) are poorly understood. Several members of the PI3K pathway, phosphatase and tensin homolog (Pten), the forkhead box, subgroup O (FOXO) transcription factors, and TSC1, have demonstrated functions in normal and leukemic stem cells but are rarely mutated in leukemia. We developed an activated allele of AKT1 that models increased signaling in normal and leukemic stem cells. In our murine bone marrow transplantation model using a myristoylated AKT1 (myr-AKT), recipients develop myeloproliferative disease, T-cell lymphoma, or AML. Analysis of the HSCs in myr-AKT mice reveals transient expansion and increased cycling, associated with impaired engraftment. myr-AKT-expressing bone marrow cells are unable to form cobblestones in long-term cocultures. Rapamycin, an inhibitor of the mammalian target of rapamycin (mTOR) rescues cobblestone formation in myr-AKT expressing bone marrow cells and increases the survival of myr-AKT mice. This study demonstrates that enhanced AKT activation is an important mechanism of transformation in AML and that HSCs are highly sensitive to excess AKT/mTOR signaling. PMID- 20008789 TI - Malignant transformation of Slp65-deficient pre-B cells involves disruption of the Arf-Mdm2-p53 tumor suppressor pathway. AB - The adapter protein Slp65 is a key component of the precursor-B (pre-B) cell receptor. Slp65-deficient mice spontaneously develop pre-B cell leukemia, but the mechanism by which Slp65(-/-) pre-B cells become malignant is unknown. Loss of Btk, a Tec-family kinase that cooperates with Slp65 as a tumor suppressor, synergizes with deregulation of the c-Myc oncogene during lymphoma formation. Here, we report that the presence of the immunoglobulin heavy chain transgene V(H)81X prevented tumor development in Btk(-/-)Slp65(-/-) mice. This finding paralleled the reported effect of a human immunoglobulin heavy chain transgene on lymphoma development in Emu-myc mice, expressing transgenic c-Myc. Because activation of c-Myc strongly selects for spontaneous inactivation of the p19(Arf) Mdm2-p53 tumor suppressor pathway, we investigated whether disruption of this pathway is a common alteration in Slp65(-/-) pre-B cell tumors. We found that combined loss of Slp65 and p53 in mice transformed pre-B cells very efficiently. Aberrations in p19(Arf), Mdm2, or p53 expression were found in all Slp65(-/-) (n = 17) and Btk(-/-)Slp65(-/-) (n = 32) pre-B cell leukemias analyzed. In addition, 9 of 10 p53(-/-)Slp65(-/-) pre-B cell leukemias manifested significant Mdm2 protein expression. These data indicate that malignant transformation of Slp65(-/ ) pre-B cells involves disruption of the p19(Arf)-Mdm2-p53 tumor suppressor pathway. PMID- 20008788 TI - HIV-1 Vpr up-regulates expression of ligands for the activating NKG2D receptor and promotes NK cell-mediated killing. AB - HIV up-regulates cell-surface expression of specific ligands for the activating NKG2D receptor, including ULBP-1, -2, and -3, but not MICA or MICB, in infected cells both in vitro and in vivo. However, the viral factor(s) involved in NKG2D ligand expression still remains undefined. HIV-1 Vpr activates the DNA damage/stress-sensing ATR kinase and promotes G(2) cell-cycle arrest, conditions known to up-regulate NKG2D ligands. We report here that HIV-1 selectively induces cell-surface expression of ULBP-2 in primary CD4(+) T lymphocytes by a process that is Vpr dependent. Importantly, Vpr enhanced the susceptibility of HIV-1 infected cells to NK cell-mediated killing. Strikingly, Vpr alone was sufficient to up-regulate expression of all NKG2D ligands and thus promoted efficient NKG2D dependent NK cell-mediated killing. Delivery of virion-associated Vpr via defective HIV-1 particles induced analogous biologic effects in noninfected target cells, suggesting that Vpr may act similarly beyond infected cells. All these activities relied on Vpr ability to activate the ATR-mediated DNA damage/stress checkpoint. Overall, these results indicate that Vpr is a key determinant responsible for HIV-1-induced up-regulation of NKG2D ligands and further suggest an immunomodulatory role for Vpr that may not only contribute to HIV-1-induced CD4(+) T-lymphocyte depletion but may also take part in HIV-1 induced NK-cell dysfunction. PMID- 20008790 TI - Stem cell exhaustion due to Runx1 deficiency is prevented by Evi5 activation in leukemogenesis. AB - The RUNX1/AML1 gene is the most frequently mutated gene in human leukemia. Conditional deletion of Runx1 in adult mice results in an increase of hematopoietic stem cells (HSCs), which serve as target cells for leukemia; however, Runx1(-/-) mice do not develop spontaneous leukemia. Here we show that maintenance of Runx1(-/-) HSCs is compromised, progressively resulting in HSC exhaustion. In leukemia development, the stem cell exhaustion was rescued by additional genetic changes. Retroviral insertional mutagenesis revealed Evi5 activation as a cooperating genetic alteration and EVI5 overexpression indeed prevented Runx1(-/-) HSC exhaustion in mice. Moreover, EVI5 was frequently overexpressed in human RUNX1-related leukemias. These results provide insights into the mechanism for maintenance of pre-leukemic stem cells and may provide a novel direction for therapeutic applications. PMID- 20008791 TI - CD137 ligand mediates opposite effects in human and mouse NK cells and impairs NK cell reactivity against human acute myeloid leukemia cells. AB - Natural killer (NK) cells play an important role in the immunosurveillance of leukemia. Their reactivity is governed by a balance of activating and inhibitory receptors including various members of the tumor necrosis factor receptor (TNFR) family. Here we report that human NK cells acquire expression of the TNFR family member CD137 upon activation, and NK cells of acute myeloid leukemia (AML) patients display an activated phenotype with substantial CD137 expression. CD137 ligand (CD137L) was detectable on leukemic cells in 35% of 65 investigated AML patients, but not on healthy CD34(+) cells, and expression was associated with monocytic differentiation. Bidirectional signaling following CD137-CD137L interaction induced the release of the immunomodulatory cytokines interleukin-10 and TNF by AML cells and directly diminished granule mobilization, cytotoxicity, and interferon-gamma production of human NK cells, which was restored by blocking CD137. Cocultures of NK cells with CD137L transfectants confirmed that human CD137 inhibits NK-cell reactivity, while activating signals were transduced by its counterpart on NK cells in mice. Our data underline the necessity to study the function of seemingly analog immunoregulatory molecules in mice compared with men and demonstrate that CD137-CD137L interaction enables immune evasion of AML cells by impairing NK-cell tumor surveillance in humans. PMID- 20008794 TI - The National Animal Germplasm Program (NAGP). PMID- 20008792 TI - STAT5 requires the N-domain for suppression of miR15/16, induction of bcl-2, and survival signaling in myeloproliferative disease. AB - Phosphorylated signal transducer and activator of transcription 5 (STAT5) is a biomarker and potential molecular target for hematologic malignancies. We have shown previously that lethal myeloproliferative disease (MPD) in mice mediated by persistently activated STAT5 (STAT5a(S711F)) requires the N-domain, but the mechanism was not defined. We now demonstrate by retrovirally complementing STAT5ab(null/null) primary mast cells that relative to wild-type STAT5a, STAT5a lacking the N-domain (STAT5aDeltaN) ineffectively protected against cytokine withdrawal-induced cell death. Both STAT5a and STAT5aDeltaN bound to a site in the bcl-2 gene and both bound near the microRNA 15b/16 cluster. However, only STAT5a could effectively induce bcl-2 mRNA and reciprocally suppress miR15b/16 leading to maintained bcl-2 protein levels. After retroviral complementation of STAT5ab(null/null) fetal liver cells and transplantation, persistently active STAT5a(S711F) lacking the N-domain (STAT5aDeltaN(S711F)) was insufficient to protect c-Kit(+)Lin(-)Sca-1(+) (KLS) cells from apoptosis and unable to induce bcl-2 expression, whereas STAT5a(S711F) caused robust KLS cell expansion, induction of bcl-2, and lethal MPD. Severe attenuation of MPD by STAT5aDeltaN(S711F) was reversed by H2k/bcl-2 transgenic expression. Overall, these studies define N-domain-dependent survival signaling as an Achilles heel of persistent STAT5 activation and highlight the potential therapeutic importance of targeting STAT5 N-domain-mediated regulation of bcl-2 family members. PMID- 20008795 TI - Two monumental, international collaborative efforts produced a consensus genetic linkage map for the chicken followed by the complete genome sequence. PMID- 20008796 TI - Influence of the breeding system on the escape response of red-legged partridges (Alectoris rufa). AB - This study aimed to determine the influence of the breeding system on the escape response of red-legged partridges comparing 147 free-born partridges captured in the wild (W) and 164 partridges from a commercial hunting farm with an intensive production system (F). All birds were individually released to the natural environment using wooden cages; the escape response was recorded with a high resolution video camera and 4 behavior parameters were analyzed: reaction time or latency, escape type, angle at the moment of flight start, and distance flown. There were significant differences on the mean reaction time depending on the origin of the partridges: 0.43 s, with a maximum of 9 s, for the W and 52.90 s, with a maximum of 120 s, in 32.3% of the F birds. Only one of the W partridges (0.68%) escaped by walking, whereas all the other W birds, and 69.5% of the F partridges, flew; the differences in the type of escape reaction between origins were significant. Considering the angle of flight start, the differences were also significant because 98.6% of W partridges showed less than 45 degrees angles, whereas 37.7% of F birds showed angles of more than 45 degrees. Thus, we can conclude that the breeding system has a great influence on the escape response of the red-legged partridges. The intensive management production systems used on the commercial game farms produced obvious changes in the escape reaction of the red-legged partridges, and this could explain the low ability of these birds to integrate and to survive in the wild due to the high predation pressure they undergo when they are used in repopulation processes. PMID- 20008797 TI - The effects of berberine on the magnitude of the acute inflammatory response induced by Escherichia coli lipopolysaccharide in broiler chickens. AB - One hundred twenty-six 19-d-old male broiler chickens were used to determine the effects of berberine on the magnitude of the acute inflammatory response induced by Escherichia coli lipopolysaccharide (LPS). The birds were weighed and randomly allotted to 1 of 3 treatments at d 19 (3 treatments x 7 replicates x 6 birds). The treatments comprised a control group in which saline was injected at d 21, an LPS-treated group in which LPS (3 mg/kg of BW) was injected at d 21, and finally a berberine and LPS-treated group in which berberine (15 mg/kg of BW) was orally administered from d 19 to d 24 with LPS injection (3 mg/kg of BW) at d 21. Injection of LPS alone decreased (P < 0.01) weight gain, feed intake, and feed conversion compared with the control and the berberine-administered group. Relative liver weight was increased (P < 0.05) in the LPS-treated group 72 h postinjection compared with the control and the berberine-treated group. Total counts of white blood cells and lymphocytes were also increased (P < 0.05) in the LPS-treated group 72 h postinjection. The heterophil concentration of the LPS treated group was greater (P < 0.05) than that of both the control and the berberine-administered group 24 h postinjection. Broilers in the LPS-treated group had greater (P < 0.05) total serum protein compared with birds in the control and the berberine-administered group both 24 and 72 h postinjection. In addition, the plasma interleukin-6 level of the LPS-treated group was significantly elevated (P < 0.01) at 24 h compared with that of the control and the berberine-administered group. Our results indicate that LPS injection initiated a series of physiological changes typical of an acute phase response in broiler chickens. These effects were largely mitigated by oral administration of berberine. PMID- 20008799 TI - Net effect of an acute phase response--partial alleviation with probiotic supplementation. AB - The acute phase response (APR) is characterized by inflammation, fever, and altered organ metabolism resulting in muscle catabolism and anorexia. Lipopolysaccharide (LPS)-induced APR may reflect depressed growth and appetite loss. Therefore, a 1-wk growth experiment was conducted to examine whether dietary supplementation of a multispecies probiotic (PoultryStar) would alleviate growth suppression and anorexia caused by LPS-induced APR. The experiment was designed with 4 treatments (n = 8 cages/treatment; 6 birds/cage) starting at 14 d of age. Before (0 to 14 d of age) and for the experiment (14 to 21 d of age), male broiler chicks were fed diets devoid of probiotic or were supplemented with 1.7 x 10(8) cfu/kg of probiotic. At 14 d of age, birds fed the diet devoid of probiotic were further divided into 3 treatments: an unchallenged positive control, LPS-challenged negative control (LPS-NC), and a treatment that was pair fed to LPS-NC. The probiotic-fed birds were also then challenged with LPS. The LPS (Escherichia coli 055:B5) was injected intraperitoneally 4 times at 48-h intervals at 1 mg/kg of BW. The LPS challenge dramatically depressed BW gain from 14 to 21 d of age by 22% (P < 0.001). However, 41% of growth depression was attributable to factors other than feed intake reduction when compared with the pair-fed treatment. Probiotic supplementation recovered 17% of depressed growth (vs. LPS-NC; P = 0.068), but this improved growth was not due to improvements in feed intake (P = 0.47). However, recovery of feed intake of the probiotic + LPS birds occurred 48-h earlier than the LPS-NC birds. Growth depression induced by LPS administration resulted in an overall relative feed intake (vs. positive control) of 0.83 and also decreased net energy and protein accretion. Probiotic supplementation did not alleviate the reduction in net energy or protein accretion induced by LPS. In conclusion, APR (induced by LPS administration) diverted a large portion of consumed nutrients from tissue accretion. Probiotic supplementation lessened the anorexic effects of LPS resulting in a trend toward BW gain improvement versus the LPS-NC. PMID- 20008798 TI - Genetic effects analysis of myeloid leukemia factor 2 and T cell receptor-beta on resistance to coccidiosis in chickens. AB - Associations between the parameters of resistance to coccidiosis and SNP in 3 candidate genes located on chromosome 1 [T cell receptor-beta (TCR-beta), myeloid leukemia factor 2 (MLF2), and lymphotactin] were determined. Single nucleotide polymorphisms were genotyped in 24 F1 generation and 290 F2 generation birds. Four SNP were identified in the lymphotactin gene, 12 were located in the TCR beta gene, and 4 in the MLF2 gene. At various times after experimental infection of the F2 generation with Eimeria maxima, BW, fecal oocyst shedding, and biochemical parameters were measured as parameters of coccidiosis resistance. Single marker association test was applied to determine the associations between the 20 SNP and the parameters of coccidiosis resistance. The maximum additive genetic effect on disease resistance of an SNP in MLF2 was explained by BW (P = 0.0002). The SNP in MLF2 significantly associated with BW was also associated with fecal oocyst shedding (P = 0.001). Four SNP associated with oocyst shedding were found within the coding region of TCR-beta (P < 0.05). Although none of the lymphotactin SNP were associated with oocyst shedding, interferon-gamma level was associated with 2 SNP in lymphotactin. These results suggest that the SNP in the MLF2 gene can be one of the markers to select coccidiosis resistance in chickens. PMID- 20008800 TI - Effect of dietary protein concentrates on the incidence of subclinical necrotic enteritis and growth performance of broiler chickens. AB - An experiment was conducted to quantify the effects of 3 nutritionally complete (similar protein and energy) corn-based diets that contained different dietary protein concentrates (potato-CP 76%, fish-CP 66%, or a mixture of soy proteins, soybean meal-CP 48%, and full-fat soy-CP 36%) on the incidence of spontaneously occurring subclinical necrotic enteritis (NE) in broiler chickens. A total of 1,260 birds were placed into 18 solid floor pens (70 birds per pen) and fed 1 of the 3 experimental diets from 15 to 31 d of age. The weight gains and feed intakes of the birds fed the potato- and fish-based diets were lower (P < 0.001) than those of the birds fed the soy-based diets. Weight gain:feed intake ratio and mortality rate were not affected (P > 0.05) by dietary treatment The birds fed the potato-based diets had a higher incidence of necrotic lesions in the duodenum (P < 0.001) and proximal jejunum (P < 0.01) than those fed the soy-based diets. The chickens fed the potato-based diet had a higher (P < 0.001) proportion of moderate to severe duodenal and distal ileal hemorrhages and liver lesions than the birds fed the soy-based diet. There was also a higher (P < 0.05) level of serum antibodies for Clostridium perfringens alpha toxin in birds fed the potato-based diet compared with the other 2 diets. The birds fed the fish-based diet had a similar (P > 0.05) incidence of subclinical NE in comparison to the birds fed the soy-based diet, although there was a higher incidence of intestinal hemorrhagic lesions. The differences in incidence of subclinical NE were not consistent with the relatively small differences in amino acid content between the diets or in the contents of nonstarch polysaccharides. However, the potato protein-based diet had higher trypsin inhibitor activity and a lower lipid content that could have contributed to the increased incidence of subclinical NE. PMID- 20008801 TI - Nutritional composition, nitrogen-corrected true metabolizable energy, and amino acid digestibilities of new corn distillers dried grains with solubles produced by new fractionation processes. AB - As ethanol production is increasing, new processes are being developed to maximize ethanol production from corn and to create new, more highly marketable corn distillers dried grains with or without solubles (DDGS or DDG, respectively). This current study evaluated coproducts produced from 2 new modified processes, the enzymatic milling (E-Mill) and the Elusieve process. The E-Mill process subjects the corn kernel to enzymes that hydrolyze starch and help in removing germ, pericarp, and endosperm fiber to create a modified higher protein, lower fiber DDG. The Elusieve process involves sieving the finished co product, DDGS, and then elutriating (air classifying) to remove fiber from the DDGS samples. A precision-fed cecectomized rooster assay was conducted to determine TME(n) and amino acid digestibilities of E-Mill DDG and several Elusieve DDGS samples produced using different screen sizes and elutriation air velocities. When compared with a conventionally processed DDGS, E-Mill DDG had increased protein (56.4 vs. 29.9%), increased TME(n) (3.656 vs. 3.299 kcal/g of DM), and higher amino acid digestibilities. When DDGS was subjected to various Elusieve processes, the resulting DDGS samples generally had an increased protein content and TME(n). As expected, the higher fiber fractions obtained from the Elusieve process had reduced protein, amino acid concentrations, amino acid digestibilities, and TME(n) in comparison to the lower fiber fractions produced from the Elusieve process. The results of this study indicate that the Elusieve and E-Mill processes can be used to increase the nutritional value of DDGS for poultry. PMID- 20008802 TI - Gompertz-Laird model prediction of optimum utilization of crude protein and metabolizable energy by French guinea fowl broilers. AB - This study was conducted to assess the influence of dietary CP and ME on growth parameters of the French guinea fowl, a meat-type variety. In a 2 x 3 x 3 factorial arrangement, 297 one-day-old French guinea keets (162 females and 135 males) were randomly assigned to experimental diets comprising 3,050, 3,100, and 3,150 kcal of ME/kg, each containing 21, 23, and 25% CP from hatch to 4 wk of age (WOA), and 3,100, 3150, and 3,200 kcal of ME/kg, each containing 19, 21, and 23% CP at 5 to 8 WOA. Using BW and G:F data from hatch to 8 WOA, the Gompertz-Laird growth model was employed to estimate growth patterns of the French guinea fowl. Mean differences in exponential growth rate, age of maximum growth, and asymptotic BW among dietary CP and ME levels were not significant. However, instantaneous growth rate and weight at inflection point were significantly higher (P < 0.05) in birds on the 25% CP diet than those on the 21% CP diet at hatch to 4 WOA (1.12 kg/wk and 0.79 kg vs. 1.04 kg/wk and 0.74 kg, respectively). The exponential growth rate was also higher (P < 0.05) in birds fed the 3,050 kcal of ME/kg diet with either 23 or 25% CP than those fed diets containing 3,050 kcal of ME/kg and 21% CP. Mean G:F was higher (P < 0.05) in birds fed diets containing 3,050 kcal of ME/kg and either 21 or 23% CP than those in other dietary treatments. Therefore, based on the Gompertz-Laird growth model estimates, feeding 21 and 23% CP and 3,100 kcal of ME/kg at hatch to 4 WOA and 19 and 21% CP with 3,150 kcal of ME/kg at 5 to 8 WOA can be recommended as adequate for growth for the French guinea fowl broilers. PMID- 20008803 TI - Effects of probiotic inclusion levels in broiler nutrition on growth performance, nutrient digestibility, plasma immunoglobulins, and cecal microflora composition. AB - The aim of this work was to investigate the effect of inclusion levels of a 5 bacterial species probiotic in broiler nutrition. Five hundred twenty-five 1-d old male Cobb broilers were allocated in 5 experimental treatments for 6 wk. The experimental treatments received a corn-soybean coccidiostat-free basal diet and depending on the addition were labeled as follows: no addition (C), 10(8) cfu probiotic/kg of diet (P1), 10(9) cfu probiotic/kg of diet (P2), 10(10) cfu probiotic/kg of diet (P3), and 2.5 mg of avilamycin/kg of diet (A). Each treatment had 3 replicates of 35 broilers each. Treatment effects on broiler growth performance and biomarkers such as ileal and total tract nutrient digestibility, plasma Ig concentration, and cecal microflora composition were determined. Differences among treatments were considered significant when P < or = 0.05. Overall BW gain was significantly higher in treatment P1 (2,293 g) compared with P2 (2,163 g), C (2,165 g), and P3 (2,167 g), with A (2,230 g) being intermediate and not different from P1. Overall feed conversion ratio values were similar and significantly better for P1 (1.80) and A (1.80) compared with P2 (1.87), C (1.89), and P3 (1.92). Ileal apparent digestibility coefficients (ADC) of CP and ether extract were higher in A. Generally, treatments A and P1 showed an improved total tract ADC for DM, organic matter, ash, ether extract, and AME(n) values. The total tract ADC of CP was higher in P1, C, and P2. There were no differences between treatments regarding plasma Ig in 14- and 42-d-old broilers. Treatments P2 and P3 were effective at beneficially modulating cecal microflora composition. In particular, the lower cecal coliform concentration (log cfu/g of wet digesta) was seen in P2 (6.12) and P3 (4.90) in 14- and 42-d old broilers, respectively, whereas at 42 d, P3 and P2 had the highest Bifidobacterium (8.31; 8.08) and Lactobacillus concentrations (8.20; 7.86), respectively. It is concluded that probiotic inclusion level had a significant effect on broiler growth responses, nutrient ADC, AME(n), and cecal microflora composition. PMID- 20008804 TI - High-throughput gene expression analysis of intestinal intraepithelial lymphocytes after oral feeding of carvacrol, cinnamaldehyde, or Capsicum oleoresin. AB - Among dietary phytonutrients, carvacrol, cinnamaldehyde, and Capsicum oleoresin are well known for their antiinflammatory and antibiotic effects in human and veterinary medicine. To further define the molecular and genetic mechanisms responsible for these properties, broiler chickens were fed a standard diet supplemented with either of the 3 phytochemicals and intestinal intraepithelial lymphocytes were examined for changes in gene expression by microarray analysis. When compared with chickens fed a nonsupplemented standard diet, carvacrol-fed chickens showed altered expression of 74 genes (26 upregulated, 48 downregulated) and cinnamaldehyde led to changes in the levels of mRNAs corresponding to 62 genes (31 upregulated, 31 downregulated). Most changes in gene expression were seen in the Capsicum-fed broilers with 98 upregulated and 156 downregulated genes compared with untreated controls. Results from the microarray analysis were confirmed by quantitative real-time PCR with a subset of selected genes. Among the genes that showed >2.0-fold altered mRNA levels, most were associated with metabolic pathways. In particular, with the genes altered by Capsicum oleoresin, the highest scored molecular network included genes associated with lipid metabolism, small molecule biochemistry, and cancer. In conclusion, this study provides a foundation to further investigate specific chicken genes that are expressed in response to a diet containing carvacrol, cinnamaldehyde, or Capsicum oleoresin. PMID- 20008805 TI - Impaired intestinal villi growth in broiler chicks with unhealed navels. AB - By the time of hatching, the residual yolk sac (RYS) should be completely internalized into the body cavity of the chick via the navel and the navel should be closed (healed). After hatching, the RYS contents are used for general growth and development including growth of the small intestine. This research examined the relationship between the presence of navel buttons (an unhealed navel covered with a scab) and the RYS weight and intestinal villi height of broiler chicks from 0 to 5 d posthatching. Hatching eggs containing live embryos were collected at 18, 19, and 20 d of incubation (n = 5/d). At 21 d of incubation, chicks with healed navels (H) and chicks with navel buttons smaller than 3 mm in diameter (B) were obtained (n = 30/group). The BW and RYS of embryos (n = 5/d) and chicks (n = 5/treatment per day) were weighed. Samples of the small intestine (duodenum and ileum) were processed for histological evaluation to measure villi height. The trial was replicated in time 4 wk after the first collection using eggs and chicks from the same flock. Data were analyzed using SAS PROC MIXED (P < or = 0.05). No significant differences in BW were observed between H and B chicks. Average RYS weight and percentage RYS were greater in B than in H chicks. The interaction of age and navel condition significantly influenced both duodenal and ileal villi heights. On most of the days, duodenal and ileal villi were shorter in B than in H chicks. The presence of small navel buttons at hatching is indicative of impaired absorption of the RYS content and decreased intestinal villi growth. Further research on the relationship between navel buttons, intestinal physiology, and yolk sac infections is advised. PMID- 20008806 TI - Adipogenesis in ducks interfered by small interfering ribonucleic acids of peroxisome proliferator-activated receptor gamma gene. AB - Peroxisome proliferator-activated receptor gamma (PPARgamma) participates in adipocyte differentiation and maintenance, including the promotion of lipid storage in mammals. In the present study, 3 duck PPARgamma small interfering RNA (siRNA) expression plasmids were constructed to investigate the effect of downregulating the expression of PPARgamma on adipogenesis and fat accumulation in ducks. The results indicate that the 3 siRNA specific for conserved regions of PPARgamma can effectively inhibit expression of PPARgamma. It was demonstrated that the expression of lipoprotein lipase and adipocyte fatty acid-binding protein in duck adipose tissue is repressed when the expression of PPARgamma is downregulated by siRNA. At the same time, the weight of abdominal fat at 21 and 35 d of age is decreased significantly (P < 0.05) compared with the control. However, the triglyceride levels in serum and muscle are not affected when the mRNA of PPARgamma is repressed. The current study indicates that the suppression of PPARgamma reduces abdominal fat deposition and regulates adipogenesis in ducks. PMID- 20008807 TI - Use of dietary thyroxine as an alternate molting procedure in spent turkey breeder hens. AB - In the turkey industry, molting is traditionally achieved by reducing photoperiod and withdrawing feed and water for several days. Although it is the most effective method, this practice is discouraged in Canada and alternative strategies need to be established. Thyroid hormone levels naturally change during molt, and dietary thyroxine (T4) supplementation was previously shown to induce molt in chickens. This study aimed at evaluating the effectiveness of supplemental dietary T4 in inducing molt in spent turkey breeder hens. One hundred twenty 75-wk-old hens were randomly divided into 4 groups (5 floor pens/replicates, 5 hens each) with the control group kept under a 14-h photoperiod and fed a breeder's diet throughout, whereas hens from the 3 other groups were supplemented with 40 ppm (45.76 mg/kg) T4 for 10 d. One treatment group was maintained under 14 h of light and fed a breeder's diet, whereas the 2 others were subjected to a drop in photoperiod to 6 h during or after supplementation and then were fed a maintenance diet. Egg production, feed intake, BW, molt, and plasma levels of T4, prolactin, and luteinizing hormone were measured. All treated hens ceased laying by d 20; however, several individuals spontaneously returned to lay when left on 14 h of light, suggesting incomplete involution of the reproductive tract. Supplementation significantly reduced feed consumption and induced rapid BW loss. All hens returned to their initial weight by the end of the experiment. Most treated hens initiated molt by d 8 of supplementation and all completed molt by d 37. Plasma T4 in treated hens increased significantly by d 3 (P < 0.05) and remained significantly higher than in controls until d 9 (P < 0.01). Levels returned to initial values by d 35. Prolactin levels did not appear to be influenced by T4 but were mainly dependent on photoperiod and reproductive stage, whereas luteinizing hormone levels remained low throughout. In summary, dietary supplementation with 40 ppm (45.76 mg/kg) T4 was successful in inducing molt in turkey breeder hens. However, dropping the photoperiod was necessary to completely reset the reproductive system. PMID- 20008808 TI - Photoperiods for broiler breeder females during the laying period. AB - Ross 308 broiler breeder females, maintained in litter-floor pens, were reared from 2 d of age on 8-h photoperiods and transferred abruptly to an 11-, 12-, 13-, or 14-h photoperiod at 20 wk of age and at a mean BW of 2.09 kg. Four qualitative dietary treatments were superimposed upon each photoperiodic treatment, but, in the absence of any significant interactions of photoperiod with diet, only photoperiodic means are reported. Age at 50% egg production (ASM) was progressively advanced as photoperiod increased from 11 to 13 h but was similar for 13-h and 14-h birds. Egg production was significantly related to ASM, with peak egg numbers being stimulated by a transfer to 13-h photoperiods. Rate of lay after 48 wk of age declined more rapidly in the 14-h birds than in any of the other groups, suggestive of an earlier onset of adult photorefractoriness. More eggs were laid outside the nest box and more cracked and dirty eggs were produced by birds on 11- and 12-h photoperiods than on 13- or 14-h photoperiods. Mean egg weight was positively correlated with ASM but negatively related to photoperiod; the number of eggs thought to be double-yolked was not significantly affected by photoperiod. Total egg mass output was not significantly affected by photoperiod. Birds exposed to 11-h and 12-h photoperiods were heavier at 60 wk than birds transferred to 13- or 14-h photoperiods. These findings show that a transfer from 8 to 13 h at 20 wk optimizes egg-laying performance in broiler breeder females, especially when they are kept beyond 60 wk of age. PMID- 20008809 TI - Effects of different acute high ambient temperatures on function of hepatic mitochondrial respiration, antioxidative enzymes, and oxidative injury in broiler chickens. AB - This study investigated the effects of different acute high ambient temperatures on dysfunction of hepatic mitochondrial respiration, the antioxidative enzyme system, and oxidative injury in broiler chickens. One hundred twenty-eight 6-wk old broiler chickens were assigned randomly to 4 groups and subsequently exposed to 25 (control), 32, 35, and 38 degrees C (RH, 70 +/- 5%) for 3 h, respectively. The rectal temperatures, activity of antioxidative enzymes (superoxide dismutase, catalase, and glutathione peroxidase), content of malondialdehyde and protein carbonyl, and the activity of mitochondrial respiratory enzymes were determined. The results showed that exposure to high ambient temperature induced a significant elevation of rectal temperature, antioxidative enzyme activity, and formation of malondialdehyde and protein carbonyl, as well as dysfunction of the mitochondrial respiratory chain in comparison with control (P < 0.05). Almost all of the indicators changed in a temperature-dependent manner with the gradual increase of ambient temperature from 32 to 38 degrees C; differences in each parameter (except catalase) among the groups exposed to different high ambient temperatures were also statistically significant (P < 0.05). The results of the present study suggest that, in the broiler chicken model used here, acute exposure to high temperatures may depress the activity of the mitochondrial respiratory chain. This inactivation results subsequently in overproduction of reactive oxygen species, which ultimately results in oxidative injury. However, this hypothesis needs to be evaluated more rigorously in future studies. It has also been shown that, with the gradual increase in temperature, the oxidative injury induced by heat stress in broiler chickens becomes increasingly severe, and this stress response presents in a temperature-dependent manner in the temperature range of 32 to 38 degrees C. PMID- 20008810 TI - Effect of glypican-1 covalently attached chains on turkey myogenic satellite cell proliferation, differentiation, and fibroblast growth factor 2 responsiveness. AB - Glypican-1 is a cell membrane heparan sulfate proteoglycan that is composed of a core protein and covalently attached glycosaminoglycan (GAG) chains and N-linked glycosylated (N-glycosylated) chains. The glypican-1 GAG chains are required for cell differentiation and responsiveness to fibroblast growth factor 2 (FGF2). The role of glypican-1 N-glycosylated chains in regulating cell activities has not been reported. The objective of the current study was to investigate the role of glypican-1 N-glycosylated chains and the interaction between N-glycosylated and GAG chains in turkey myogenic satellite cell proliferation, differentiation, and FGF2 responsiveness. The wild-type turkey glypican-1 and turkey glypican-1 with mutated GAG chain attachment sites were cloned into the pCMS-EGFP mammalian expression vector and were used as templates to generate glypican-1 N glycosylated 1-chain and no-chain mutants with or without GAG chains by site directed mutagenesis. The wild-type glypican-1 and all glypican-1 N-glycosylated 1-chain and no-chain mutants with or without GAG chains were transfected into turkey myogenic satellite cells. Cell proliferation, differentiation, and FGF2 responsiveness were measured. The overexpression of glypican-1 N-glycosylated 1 chain and no-chain mutants without GAG chains increased cell proliferation and differentiation compared with the wild-type glypican-1 but not the glypican-1 N glycosylated mutants with GAG chains attached. Cells overexpressing glypican-1 N glycosylated mutants with or without GAG chains increased cell responsiveness to FGF2 compared with wild-type glypican-1. These data suggest that glypican-1 N glycosylated chains and GAG chains are critical in regulating turkey myogenic satellite cell proliferation, differentiation, and responsivness to FGF2. PMID- 20008811 TI - Autonomic control of heart rate and its variability during normoxia and hypoxia in emu (Dromaius novaehollandiae) hatchlings. AB - Heart rate variability is a common feature of the vertebrate cardiovascular system and is a consequence of variable input from the sympathetic and parasympathetic branches of the autonomic nervous system. The aim of this study was to examine the role of the autonomic nervous system in regulating heart rate and heart rate variability in 1-d-old emu hatchlings in normoxia and during exposure to 10% O2. The role of the autonomic nervous system in controlling emu heart rate and its variability was examined by blocking the action of the cholinergic and adrenergic pathways by administration of atropine and propranolol. Heart rate of 1-d-old hatchlings exhibited a significant cholinergic tone of 60 +/- 22 beats per minute (bpm) and beta-adrenergic tone of 28 +/- 17 bpm. Cholinergic tone was unchanged during hypoxic exposure (63.5 +/- 17.7 bpm), but adrenergic tone doubled to 68 +/- 15 bpm. Initially, the majority of hatchlings exhibited high frequency oscillations with a spectral peak at 0.22 +/- 0.02 Hz, associated with respiratory sinus arrhythmia. Beta-adrenergic blockade had no effect on respiratory sinus arrhythmia or spectral power in high frequency (HF; 0.1 to 0.7 Hz), low frequency (LF; 0.01 to 0.1 Hz), or total frequency (TF) ranges. After cholinergic blockade, spectral power in HF, LF, and TF ranges and respiratory sinus arrhythmia were all abolished. Hypoxia did not initially alter spectral power in the HF, LF, or TF ranges. beta-Adrenergic blockade along with hypoxia produced a significant increase in LF oscillations. A distinct LF oscillation appeared in most birds exposed to hypoxia that was abolished by cholinergic blockade. We conclude that although both the sympathetic and parasympathetic system exert a tonic influence on heart rate, the majority of HF and TF heart rate variability is mediated by the parasympathetic system in the emu hatchling. The sympathetic system contributes to LF heart rate oscillations by suppressing the influence of the parasympathetic system on LF oscillations. PMID- 20008812 TI - Microbiological and chemical analyses of ice collected from a commercial poultry processing establishment. AB - A study was conducted to evaluate the microbiological and chemical characteristics of ice collected from a commercial poultry further processing facility. During each of 3 visits, the following ice samples were collected: 1) freshly prepared, unused ice; 2) product-contact ice from ice-packed poultry parts; 3) product-contact ice from ice-packed poultry that had been visibly inspected and condemned as not for reuse; and 4) product-contact ice from ice packed poultry that had passed visible inspection and had been prepared for reuse by washing (rinse with potable water and drain). The overall pattern for lowest to highest numbers of total aerobic microorganisms, coliforms, Escherichia coli, and Enterobacteriaceae was as follows: unused ice < washed ice < product-contact ice < condemned ice. Mean levels of total aerobic microorganisms, coliforms, and Enterobacteriaceae in the unused ice were 0.3, 0.4, and 0.4 log10 cfu/mL, respectively. No E. coli was detected in the unused or washed ice, and levels were 0.5 and 1.5 log10 cfu/mL in the product-contact and condemned ice samples, respectively. Mean levels of bacteria enumerated in condemned ice were 0.8, 1.0, and 0.6 log10 cfu/mL higher than the levels of bacteria found in product-contact ice for coliforms, E. coli, and Enterobacteriaceae, respectively. Washing and draining the product-contact ice decreased counts by 0.9, 0.7, 0.5, and 1.7 log10 cfu/mL for total aerobic microorganisms, coliforms, E. coli, and Enterobacteriaceae, respectively. All of the ice samples had similar pH values (pH 6.1 to 6.4). Unused and washed ice were not significantly different for total solids, total suspended solids, total Kjeldahl nitrogen, and chemical oxygen demand. Condemned ice contained the highest concentration of total solids, total suspended solids, total Kjeldahl nitrogen, and chemical oxygen demand, with levels more than 3 times that found in product contact ice. Data from the present study demonstrate that visible contamination in ice corresponds with increased microbiological and chemical contamination. Product-contact ice may be washed and the washing procedure can reduce the bacterial, solids, nitrogen, and organic loads. PMID- 20008813 TI - Comparison of algal and fish sources on the oxidative stability of poultry meat and its enrichment with omega-3 polyunsaturated fatty acids. AB - Human consumption of long-chain n-3 polyunsaturated fatty acids (LC n-3 PUFA) is below recommendations, and enriching chicken meat (by incorporating LC n-3 PUFA into broiler diets) is a viable means of increasing consumption. Fish oil is the most common LC n-3 PUFA supplement used but is unsustainable and reduces the oxidative stability of the mat. The objective of this experiment was to compare fresh fish oil (FFO) with fish oil encapsulated (EFO) in a gelatin matrix (to maintain its oxidative stability) and algal biomass at a low (LAG, 11), medium (MAG, 22), or high (HAG, 33 g/kg of diet) level of inclusion. The C22:6n-3 contents of the FFO, EFO, and MAG diets were equal. A control (CON) diet using blended vegetable oil was also made. As-hatched 1-d-old Ross 308 broilers (144) were reared (21 d) on a common starter diet then allocated to treatment pens (4 pens per treatment, 6 birds per pen) and fed treatment diets for 21 d before being slaughtered. Breast and leg meat was analyzed (per pen) for fatty acids, and cooked samples (2 pens per treatment) were analyzed for volatile aldehydes. Concentrations (mg/100 g of meat) of C20:5n-3, C22:5n-3, and C22:6n-3 were (respectively) CON: 4, 15, 24; FFO: 31, 46, 129; EFO: 18, 27, 122; LAG: 9, 19, 111; MAG: 6, 16, 147; and HAG: 9, 14, 187 (SEM: 2.4, 3.6, 13.1) in breast meat and CON: 4, 12, 9; FFO: 58, 56, 132; EFO: 63, 49, 153; LAG: 13, 14, 101; MAG: 11, 15, 102; HAG: 37, 37, 203 (SEM: 7.8, 6.7, 14.4) in leg meat. Cooked EFO and HAG leg meat was more oxidized (5.2 mg of hexanal/kg of meat) than the other meats (mean 2.2 mg/kg, SEM 0.63). It is concluded that algal biomass is as effective as fish oil at enriching broiler diets with C22:6 LC n-3 PUFA, and at equal C22:6n-3 contents, there is no significant difference between these 2 supplements on the oxidative stability of the meat that is produced. PMID- 20008814 TI - The effects of commercial cool water washing of shell eggs on Haugh unit, vitelline membrane strength, aerobic microorganisms, and fungi. AB - Current egg washing practices use wash water temperatures averaging 49 degrees C and have been found to increase internal egg temperature by 6.7 to 7.8 degrees C. These high temperatures create a more optimal environment for bacterial growth, including Salmonella Enteritidis if it is present. Salmonella Enteritidis is the most common human pathogen associated with shell eggs and egg products. Its growth is inhibited at temperatures of 7.2 degrees C and below. The objective of this study was to determine if commercially washing eggs in cool water would aid in quickly reducing internal egg temperature, preserving interior egg quality, and slowing microbial growth. During 3 consecutive days, eggs were washed using 4 dual-tank wash water temperature schemes (HH = 49 degrees C, 49 degrees C; HC = 49 degrees C, 24 degrees C; CC = 24 degrees C, 24 degrees C; CH = 24 degrees C, 49 degrees C) at 2 commercial processing facilities. A 10-wk storage study followed, in which vitelline membrane strength, Haugh unit, and aerobic microorganisms and fungi (yeasts and molds) were monitored weekly. As storage time progressed, average Haugh unit values declined 14.8%, the average force required to rupture the vitelline membrane decreased 20.6%, average numbers of bacteria present on shell surfaces decreased 11.3%, and bacteria present in egg contents increased 39.5% during storage. Wash water temperature did not significantly affect Haugh unit values, vitelline membrane strength, or the numbers of aerobic microorganisms and fungi within the shell matrices of processed eggs. Results of this study indicate that incorporating cool water into commercial shell egg processing, while maintaining a pH of 10 to 12, lowers postprocessing egg temperatures and allows for more rapid cooling, without causing a decline in egg quality or increasing the presence of aerobic microorganisms and fungi for approximately 5 wk postprocessing. PMID- 20008815 TI - Sampling method and location affect recovery of coliforms and Escherichia coli from broiler carcasses. AB - Two experiments were conducted, the first to determine whether numbers of recovered bacteria differed due to sampling method used or due to location on carcass sampled (breast or leg quarters) and the second to determine if numbers of bacteria differed between the front (ventral) and back (dorsal) side of the carcass. In both experiments, eviscerated broiler carcasses were obtained from a commercial processing plant just before the final inside-outside bird washer. In experiment 1, carcasses (3 in each of 4 replicate trials) were separated into leg quarters and breast quarters (n = 48) and either rinsed or ground and stomached for microbiological sampling. In experiment 2, for 3 replicate trials of 4 carcasses each, necks, wings, and legs were manually removed; the remaining trunks were cut through the sides to produce front (ventral) and back (dorsal) halves (n = 24); and then rinsed. For both experiments, coliforms and Escherichia coli were enumerated. In experiment 1, significantly higher numbers (P < 0.05) of coliforms and E. coli were recovered by rinsing than by grinding from both breast and leg quarters. Leg quarters were found to have higher bacterial numbers than breasts from grind samples, but no quarter differences were found for rinse samples. In experiment 2, higher (P < 0.05) numbers of coliforms and E. coli were recovered from the dorsal carcass half compared with the ventral half. Bacterial counts of broiler carcasses are affected by both the sampling method used and by carcass location sampled. PMID- 20008816 TI - Growth analysis of chickens fed diets varying in the percentage of metabolizable energy provided by protein, fat, and carbohydrate through artificial neural network. AB - A radial basis function neural network (RBFN) approach was used to develop a multi-input, multi-output model for the effect of diets varying in the percentage of ME provided by protein (% ME(P)), fat (% ME(F)), and carbohydrate (% ME(C)) on live weight gain, protein gain, and fat gain in growing chickens. Thirty-three data lines representing response of the White Leghorn male chickens during 23 to 33 d of age to the diets varying in the % ME(P), % ME(F), and % ME(C) were obtained from literature and used to train the RBFN model. The prediction values of the RBFN model were compared with those obtained by multiple regression models to assess the fitness of these 2 methods. The fitness of the models was tested using R2, MS error, mean absolute deviation, residual SD, and bias. The developed RBFN model was used to evaluate the relative importance of each input parameter on chicken growth using a sensitivity analysis method. The calculated statistical values corresponding to the RBFN model showed a higher accuracy of prediction than multiple regression models. The sensitivity analysis on the model indicated that dietary % ME(P) is the most important variable in the growth of chickens, followed by dietary % ME(F) and % ME(C). It was found that the RBFN model is an appropriate tool to recognize the patterns of input-output data or to predict chicken growth in terms of live weight gain, protein gain, and fat gain given the proportion of dietary percentage of ME intake supplied through protein, fat, or carbohydrates. PMID- 20008817 TI - Veterinary pharmaceuticals and antibiotic resistance of Escherichia coli isolates in poultry litter from commercial farms and controlled feeding trials. AB - Veterinary pharmaceuticals are commonly used in poultry farming to prevent and treat microbial infections as well as to increase feed efficiency, but their use has created public and environmental health concerns. Poultry litter contains antimicrobial residues and resistant bacteria; when applied as fertilizer, the level and effects of these pharmaceuticals and antimicrobial-resistant bacteria in the environment are of concern. The purpose of this study was to investigate poultry litter for veterinary pharmaceuticals and resistance patterns of Escherichia coli. Litter samples were collected from controlled feeding trials and from commercial farms. Feed additives bacitracin, chlortetracycline, monensin, narasin, nicarbazin, penicillin, salinomycin, and virginiamycin, which were present in the feed on commercial farms and added to the feed in the controlled trials, were extracted in methanol and analyzed by liquid chromatography-mass spectrometry techniques. Sixty-nine E. coli were isolated and identified by API 20E. The susceptibility of the isolates to antibiotics was determined using Avian plates and the Sensititer automated system. This study confirmed the presence of antimicrobial residues in broiler litter from controlled environments as well as commercial farms, ranging from 0.07 to 66 mg/L depending on the compound. Concentrations of individual residues were higher in litter from controlled feeding trials than those from commercial farms. All E. coli isolates from commercial farms were multiresistant to at least 7 antibiotics. Resistance to beta-lactam antibiotics (amoxicillin, ceftiofur), tetracyclines, and sulfonamides was the most prevalent. This study concluded that broiler litter is a source of antimicrobial residues and represents a reservoir of multiple antibiotic-resistant E. coli. PMID- 20008818 TI - Comparative field evaluations of in ovo applied technology. AB - Mass vaccination of poultry has advanced in recent years with field application of in ovo vaccination technologies in hatcheries worldwide. Working from a laboratory concept to a commercially available system within a matter of a decade, with subsequent full application to 90% of US broiler production a few years later, the commercial systems continue to be an area for study and continued understanding. Within the production setting, 2 methods to evaluate the types of equipment available for this technology were evaluated in this research. Utilizing a whole hatchery approach and paired testing in a field setting allowed for direct comparison of the 2 US-manufactured in ovo systems (Embrex Inovoject System and Intelliject). The results of the whole hatchery study showed an advantage for 1 system at d 18 and 19 transfer, with the Embrex Inovoject System showing as much as a 2.32% advantage in percentage of hatch. Paired testing showed a statistical difference in percentage of hatchability at d 18 of transfer favoring the Embrex Inovoject System but no difference at d 19 transfer. Differences in the 2 systems were additionally evaluated in terms of day of transfer and flock age. PMID- 20008819 TI - Preclinical acute toxicology study of surfactant-stabilized ultrasound contrast agents in adult rats. AB - Gas-filled microbubbles are used as contrast agents in diagnostic ultrasound imaging. A preclinical, acute toxicity study of 2 surfactant-stabilized ultrasound contrast agents (ST68 and ST44) was conducted. Subjects were 104 Sprague-Dawley rats (experimental doses, 0.1, 0.2, 0.8, and 1.0 mL/kg; control, 1.0 mL/kg saline) that were studied for 14 days after contrast; clinical signs, weight, blood, and urine were evaluated. Histopathology was performed following euthanasia. Of the 40 animals receiving ST44, 4 died prematurely and a dose dependency was demonstrated (P = .011), whereas in the ST68 groups only 1 death occurred (no dose dependency; P = .48). Only the weight of rats injected with ST44 varied significantly (P = .0003). This dependency was also found for 3 of 5 urine parameters and 4 of 36 blood parameters (P < .05). For ST68, only 1 urine parameter showed significance (P < .0001). Giant cell infiltration in the lungs was significantly higher than controls in the ST44 0.1 mL/kg and the ST68 0.8-1.0 mL/kg groups (P < .01). It is concluded that the prudent choice for future nonrodent, toxicology studies and potentially for human clinical trials is ST68 (given the deaths in the ST44 groups). PMID- 20008821 TI - The gold medal face of ADHD. PMID- 20008820 TI - Effects of dance on gait and balance in Parkinson's disease: a comparison of partnered and nonpartnered dance movement. AB - Partnered tango dance can improve balance and gait in individuals with Parkinson's disease (PD). Partnered dance may allow these individuals to challenge balance more than nonpartnered dance. Alternatively, partnered practice could reduce balance gains because the participant may rely on the partner as a balance aid when challenged. The authors compared the effects of partnered and nonpartnered dance on balance and mobility in 39 people (11 women) with mild moderate PD (Hoehn and Yahr stages I-III). Participants were randomly assigned to partnered or nonpartnered tango and attended 1-hour classes twice per week, completing 20 lessons within 10 weeks. Balance and gait were evaluated in the weeks immediately before, immediately after, and 1 month after the intervention. Both groups significantly improved on the Berg Balance Scale, comfortable and fast-as-possible walking velocity, and cadence. Improvements were maintained at the 1-month follow-up. The nonpartnered class improved as much as the partnered class; however, partnered participants expressed more enjoyment and interest in continuing. PMID- 20008823 TI - The stress process in palliative cancer care: a qualitative study on informal caregiving and its implication for the delivery of care. AB - OBJECTIVE: The primary objective of this study was to examine how the comprehensive nature of the Stress Process Model could elucidate on the stressors associated with caring for a palliative cancer patient. METHOD: A qualitative research strategy involving home-based face-to-face interviews with 12 bereaved family caregivers was used to examine the caregiving experience. RESULTS: The primary stressors associated with caring for the palliative cancer care patients stemmed from care recipient symptoms and personal care needs. The absence of adequate support from the formal health care delivery system was a consistent message from all participants. There was evidence of financial stress primarily associated with the purchase of private home care to supplement formal care. In contrast, the resources that family caregivers relied on to moderate the stressful effects of caregiving included extended family, friends, and neighbors. While the stress of direct caregiving was high, the study revealed that formal care was also a significant source of stress for family caregivers. CONCLUSION: It was concluded that an appropriately financed, integrated system of care that followed a person-centered philosophy of care would best meet the needs of the patient and his or her family. PMID- 20008822 TI - Anterior cingulate volumetric alterations in treatment-naive adults with ADHD: a pilot study. AB - OBJECTIVE: We sought to examine preliminary results of brain alterations in anterior cingulate cortex (ACC) in treatment-naive adults with ADHD. The ACC is a central brain node for the integration of cognitive control and allocation of attention, affect and drive. Thus its anatomical alteration may give rise to impulsivity, hyperactivity and inattention, which are cardinal behavioral manifestations of ADHD. METHOD: Segmentation and parcellation of the ACC was performed on controls (n = 22), treated (n = 13) and treatment-naive adults with ADHD (n = 13). RESULTS: There was a 21% volume reduction in the left ACC of the treatment-naive group relative to the control group. Also, there was a 23% volume reduction in the right ACC of the treated group relative to the control group. CONCLUSION: These results raise the possibility that in ADHD there are volumetric deficits persistent into adulthood, that are independent of medical treatment. PMID- 20008824 TI - Intermittent cancer pain: clinical importance and an updated cancer pain classification. AB - AIM: We report the characteristics of intermittent cancer pain. In addition, we propose a new clinically based classification. METHODS: Consecutive patients with cancer referred to our palliative medicine service were consented and underwent a comprehensive pain evaluation including available laboratory and radiological studies, at the time of initial contact. RESULTS AND DISCUSSION: In total, 100 consecutive patients reported 158 different pain sites. Pain temporal pattern observed was 60% of patients had continuous (CP) plus intermittent pain (IP); 29% IP alone; and 11% CP alone. The etiology of IP was somatic (58%), visceral (24%), neuropathic (7%), and mixed (11%). Median duration of IP was 4 months with a median daily frequency of 4 episodes. Consequently, we propose that IP be classified into IP alone or nonbreakthrough pain (NBP; because there is no underlying CP or around-the-clock [ATC] opioids used) and breakthrough pain (BP; because there is underlying CP or/and ATC opioids used). We propose that both BP and NBP be each subclassified into 3 categories: (1) incident, (2) non-incident, and (3) mixed. In addition, a 4th category exclusive to BP: end-of-dose failure. Incident pains made up (N = 42, 47%) nearly half of all IP. According to our classification, incident pain was part of BP in 41% (N = 25) or NBP in 58% (N = 17). Incident NBP received less treatment than incident BP, and it was less controlled. CONCLUSION: (1) Intermittent pain is a major problem in patients with cancer, (2) NBP is a common but under-recognized form of cancer pain, (3) NBP is less defined and controlled than BP, (4) incident NBP accounts for 40% of all incident cancer pain, and (5) variable IP definitions and classifications make comparisons between studies difficult. PMID- 20008825 TI - Continuous deep sedation until death: palliation or physician-assisted death? AB - Published literature has not discerned end-of-life palliative versus life shortening effects of pharmacologically maintaining continuous deep sedation until death (i.e., dying in deep sleep) compared with common sedation practices relieving distress in the final conscious phase of dying. Continuous deep sedation predictably suppresses brainstem vital centers and shortens life. Continuous deep sedation remains controversial as palliation for existential suffering and in elective death requests by discontinuation of chronic ventilation or circulatory support with mechanical devices. Continuous deep sedation contravenes the double-effect principle because: (1) it induces permanent coma (intent of action) for the contingency relief of suffering and for social isolation (desired outcomes) and (2) because of its predictable and proportional life-shortening effect. Continuous deep sedation should be distinguished from common sedation practices for palliation and characterized instead as physician-assisted death. PMID- 20008826 TI - Alterations in the modulation of cerebrovascular tone and blood flow by nitric oxide synthases in SHRsp with stroke. AB - AIMS: The modulation of myogenic function and cerebral blood flow (CBF) by nitric oxide (NO) synthases (NOS) was assessed in the middle cerebral arteries (MCAs) of Kyoto Wistar stroke prone hypertensive rats (SHRsp) in relation to haemorrhagic stroke development. METHODS AND RESULTS: MCAs were studied with a pressure myograph. CBF in MCA perfusion domain was measured using laser Doppler techniques. NOS isozymes were identified using immunohistochemistry. MCAs expressed endothelial, neuronal, and inducible NOS (eNOS, nNOS, and iNOS, respectively) in the endothelium, nNOS and traces of iNOS in smooth muscle and adventitial cells. Before stroke, MCA pressure-dependent constriction (PDC) was superimposed over basal non-pressure-dependent tone (BNPDT). Endothelial NO generation and non-endothelial nNOS but not iNOS reduced BNPDT and increased the lumen diameter at which PDC initiated without altering the amplitude of PDC. NOS inhibition decreased CBF and increased the upper blood pressure limit of autoregulation. PDC, CBF autoregulation, and NOS dilatory influence were lost, and BNPDT was increased in MCAs from SHRsp with stroke. The expression of NOS isozymes and MCA reactivity to NO donors was not altered. NOS activity was not recovered by in vitro l-arginine or tetrahydrobiopterin supplementation, l arginase inhibition or superoxide scavengers. CONCLUSION: The loss of PDC and CBF autoregulation during hypertension may facilitate over-perfusion and cerebral haemorrhage formation in SHRsp. NOS dysfunction in MCAs preceded stroke and involved the inactivation of eNOS and nNOS in areas not subjected to hyper distension. The elevation in BNPDT due to NOS inactivation may oppose over perfusion in the absence of CBF autoregulation. PMID- 20008827 TI - Measuring intracellular pH in the heart using hyperpolarized carbon dioxide and bicarbonate: a 13C and 31P magnetic resonance spectroscopy study. AB - AIMS: Technological limitations have restricted in vivo assessment of intracellular pH (pH(i)) in the myocardium. The aim of this study was to evaluate the potential of hyperpolarized [1-(13)C]pyruvate, coupled with (13)C magnetic resonance spectroscopy (MRS), to measure pH(i) in the healthy and diseased heart. METHODS AND RESULTS: Hyperpolarized [1-(13)C]pyruvate was infused into isolated rat hearts before and immediately after ischaemia, and the formation of (13)CO(2) and H(13)CO(3)(-) was monitored using (13)C MRS. The HCO(3)(-)/CO(2) ratio was used in the Henderson-Hasselbalch equation to estimate pH(i). We tested the validity of this approach by comparing (13)C-based pH(i) measurements with (31)P MRS measurements of pH(i). There was good agreement between the pH(i) measured using (13)C and (31)P MRS in control hearts, being 7.12 +/- 0.10 and 7.07 +/- 0.02, respectively. In reperfused hearts, (13)C and (31)P measurements of pH(i) also agreed, although (13)C equilibration limited observation of myocardial recovery from acidosis. In hearts pre-treated with the carbonic anhydrase (CA) inhibitor, 6-ethoxyzolamide, the (13)C measurement underestimated the (31)P measured pH(i) by 0.80 pH units. Mathematical modelling predicted that the validity of measuring pH(i) from the H(13)CO(3)(-)/(13)CO(2) ratio depended on CA activity, and may give an incorrect measure of pH(i) under conditions in which CA was inhibited, such as in acidosis. Hyperpolarized [1-(13)C]pyruvate was also infused into healthy living rats, where in vivo pH(i) from the H(13)CO(3)( )/(13)CO(2) ratio was measured to be 7.20 +/- 0.03. CONCLUSION: Metabolically generated (13)CO(2) and H(13)CO(3)(-) can be used as a marker of cardiac pH(i) in vivo, provided that CA activity is at normal levels. PMID- 20008828 TI - Cost-benefit analysis of supplemented very low-protein diet versus dialysis in elderly CKD5 patients. AB - BACKGROUND: Dialysis increases patient life expectancy but is associated with clinically severe and costly complications. Health and economic benefits could derive from postponing dialysis with a supplemented very low-protein diet (sVLPD). METHODS: An economic evaluation was conducted to compare benefits and costs of sVLPD versus dialysis in elderly CKD5 patients. Data from 57 patients aged >or=70 years, with glomerular filtration rate (GFR) 5-7 mL/min, previously participating in a clinical trial demonstrating non-inferior mortality and morbidity of starting sVLPD compared to dialysis treatment, were analysed: 30 patients were randomized to dialysis and 27 to sVLPD. A cost-benefit analysis was conducted, in the perspective of the National Health Service (NHS). Direct medical and non-medical benefits and costs occurring in 3.2 mean years of follow up were quantified: time free from dialysis, cost of dialysis treatment, hospitalization, drugs, laboratory/instrumental tests, medical visits and travel and energy consumption to receive dialysis. Prices/tariffs valid in 2007 were used, with an annual discount rate of 5% applied to benefits and costs occurring after the first year. Sensitivity analyses were conducted to identify how estimates could vary in different contexts of applications. Results are reported as net benefit, expressed as mean euro/patient (patient-year). RESULTS: The opportunity to safely postpone initiation of dialysis of 1 year/patient on average translated into an economic benefit to the NHS, corresponding to 21 180 euro/patient in the first, 6500 euro/patient in the second and 682 euro/patient in the third year of treatment, with a significant net benefit in favour of sVLPD even in a worst-case hypothesis. CONCLUSION: The initiation of sVLPD in elderly CKD5 subjects is a safe and beneficial strategy for these patients and allows them to gain economic resources that can be allocated to further health care investments. PMID- 20008829 TI - Albuminuria, impaired kidney function and cardiovascular outcomes or mortality in the elderly. AB - BACKGROUND: Kidney disease is a risk factor for mortality and cardiovascular disease in older adults, but the separate and combined effects of albuminuria and cystatin C, a novel marker of glomerular filtration, are not known. METHODS: We examined associations of these markers with mortality and cardiovascular outcomes during a median follow-up of 8.3 years in 3291 older adults in the Cardiovascular Health Study. Kidney disease was assessed using urinary albumin/creatinine ratio (ACR), cystatin C and Modification of Diet in Renal Disease estimated glomerular filtration rate (eGFR). We defined subgroups based on presence of microalbuminuria (MA, ACR > 30 mg/g) and categories of normal kidney function (cystatin C < 1.0 mg/L and eGFR > 60 mL/min/1.73 m(2)); preclinical kidney disease (cystatin C level > 1.0 mg/l but eGFR > 60 mL/min/1.73 m(2)); and chronic kidney disease (CKD) (eGFR < 60 mL/min/1.73 m(2)). Cox proportional hazards models were used to examine associations between these six subgroups and all cause or cardiovascular mortality, myocardial infarction and heart failure. RESULTS: One thousand one hundred fifty (34.9%) had normal kidney function (12.2% with MA), 1518 (46.1%) had preclinical kidney disease (17.9% with MA) and 622 (18.9%) had CKD (47% with MA). After adjustment, the presence of either preclinical kidney disease or MA was associated with an over 50% increase in mortality risk; the presence of both was associated with a 2.4-fold mortality risk. Those with CKD and MA were at highest risk, with a nearly 4-fold mortality risk. CONCLUSION: Elevated cystatin C and albuminuria are common, identify different subsets of the older population, and are independent, graded risk factors for cardiovascular disease and mortality. PMID- 20008830 TI - Costs of dialysis--a regional population-based analysis. AB - BACKGROUND: Population-based estimates of costs of renal replacement therapy are scarce in the literature. The aim of our study was to calculate the costs of long term dialysis in 2006 on the basis of patient-specific data from a well-defined population in a region in western Germany (n = 310,757). METHODS: Cost estimation was performed from the perspective of the statutory health insurance. All dialysis patients from the study region (n = 344, 54% male, mean age (+/-SD) 69 +/- 13 years, 42% diabetic) were assessed for the costs of the dialysis procedures, dialysis-related hospital admissions, outpatient contacts outside of our dialysis center, dialysis-related medication, patient transportation and related costs (e.g. reimbursement fees on the basis of the German diagnosis related group system, price scales). We estimated the cumulative cost per patient year in 2006 (in Euros), along with the 10th and 90th percentiles and the 95% confidence intervals (CI) by using bootstrapping procedures. RESULTS: The mean total dialysis-related cost in 2006 was 54,777 Euros (95% CI, 51,445-65,705) per patient year. The largest part of the costs (55%) was caused by the dialysis procedures, followed by the costs of medication (22%), hospitalization (14%) and transportation (8%). The total cost increased significantly with increasing age. No significant association was found between total cost and sex, dialysis strategy, end-stage renal disease duration and diabetes. CONCLUSIONS: We present for the first time a cost estimation of dialysis in Germany on the basis of patient-level data in a population-based sample. Except age, patient characteristics were not significantly associated with costs. The largest part of the costs was caused by the dialysis procedures themselves; however, other dialysis-specific health care utilization also strongly contributed to the total cost. PMID- 20008832 TI - Threshold in stage-specific embryonic glycotypes uncovered by a full portrait of dynamic N-glycan expression during cell differentiation. AB - Although various glycoforms appear to participate independently in multiple molecular interactions in cellular adhesion that contribute to embryogenesis and organogenesis, a full portrait of the glycome diversity and the effect of the structural variations of cellular glycoforms on individual cell stages in proliferation and differentiation remain unclear. Here we describe a novel concept for the characterization of dynamic glycoform alteration during cell differentiation by means of "glycoblotting-based cellular glycomics," the only method allowing for rapid and quantitative glycan analysis. We demonstrated that processes of dynamic cellular differentiation of mouse embryonic carcinoma cells, P19CL6 and P19C6, and mouse embryonic stem cells into cardiomyocytes or neural cells can be monitored and characterized quantitatively by profiling entire N glycan structures of total cell glycoproteins. Whole N-glycans enriched and identified by the glycoblotting method (67 glycans for P19CL6, 75 glycans for P19C6, and 72 glycans for embryonic stem cells) were profiled and bar-coded quantitatively with respect to the ratio of subgroups composed of characteristic glycoforms, namely glycotypes. PMID- 20008831 TI - Effect of apolipoprotein-B synthesis inhibition on liver triglyceride content in patients with familial hypercholesterolemia. AB - To investigate the impact of mipomersen, an apolipoprotein B-100 (apoB) synthesis inhibitor, on intra-hepatic triglyceride content (IHTG content), we conducted a randomized, double-blind, placebo-controlled study in 21 patients with familial hypercholesterolemia (FH). Subjects received a weekly subcutaneous dose of 200 mg mipomersen or placebo for 13 weeks while continuing conventional lipid lowering therapy. The primary endpoint was change in IHTG content from week 0 to week 15 as measured by localized proton magnetic resonance spectroscopy (1H-MRS). Thirteen weeks of mipomersen administration reduced LDL-cholesterol by 22.0 (17.8) % and apoB by 19.9 (17.4) % (both P < 0.01). One of 10 patients (10%) in the mipomersen-treated group developed mild hepatic steatosis at week 15, which was reversible following mipomersen discontinuation. For the group, there was a trend toward an increase in IHTG content [placebo; baseline: 1.2% and week 15: 1.1%; change -0.1 (0.9). Mipomersen; baseline: 1.2% and week 15: 2.1%; change 0.8 (1.7) (P = 0.0513)]. Mipomersen administration for 13 weeks to subjects with FH is associated with a trend toward an increase in IHTG content. Future studies evaluating the effects of long-term use of mipomersen reaching more profound reductions in apoB are required prior to broader use of this compound. PMID- 20008833 TI - The nutrigenetics of hyperhomocysteinemia: quantitative proteomics reveals differences in the methionine cycle enzymes of gene-induced versus diet-induced hyperhomocysteinemia. AB - Hyperhomocysteinemia has long been associated with atherosclerosis and thrombosis and is an independent risk factor for cardiovascular disease. Its causes include both genetic and environmental factors. Although homocysteine is produced in every cell as an intermediate of the methionine cycle, the liver contributes the major portion found in circulation, and fatty liver is a common finding in homocystinuric patients. To understand the spectrum of proteins and associated pathways affected by hyperhomocysteinemia, we analyzed the mouse liver proteome of gene-induced (cystathionine beta-synthase (CBS)) and diet-induced (high methionine) hyperhomocysteinemic mice using two-dimensional difference gel electrophoresis and Ingenuity Pathway Analysis. Nine proteins were identified whose expression was significantly changed by 2-fold (p < or = 0.05) as a result of genotype, 27 proteins were changed as a result of diet, and 14 proteins were changed in response to genotype and diet. Importantly, three enzymes of the methionine cycle were up-regulated. S-Adenosylhomocysteine hydrolase increased in response to genotype and/or diet, whereas glycine N-methyltransferase and betaine homocysteine methyltransferase only increased in response to diet. The antioxidant proteins peroxiredoxins 1 and 2 increased in wild-type mice fed the high methionine diet but not in the CBS mutants, suggesting a dysregulation in the antioxidant capacity of those animals. Furthermore, thioredoxin 1 decreased in both wild-type and CBS mutants on the diet but not in the mutants fed a control diet. Several urea cycle proteins increased in both diet groups; however, arginase 1 decreased in the CBS(+/-) mice fed the control diet. Pathway analysis identified the retinoid X receptor signaling pathway as the top ranked network associated with the CBS(+/-) genotype, whereas xenobiotic metabolism and the NRF2 mediated oxidative stress response were associated with the high methionine diet. Our results show that hyperhomocysteinemia, whether caused by a genetic mutation or diet, alters the abundance of several liver proteins involved in homocysteine/methionine metabolism, the urea cycle, and antioxidant defense. PMID- 20008834 TI - Analysis of human C1q by combined bottom-up and top-down mass spectrometry: detailed mapping of post-translational modifications and insights into the C1r/C1s binding sites. AB - C1q is a subunit of the C1 complex, a key player in innate immunity that triggers activation of the classical complement pathway. Featuring a unique structural organization and comprising a collagen-like domain with a high level of post translational modifications, C1q represents a challenging protein assembly for structural biology. We report for the first time a comprehensive proteomics study of C1q combining bottom-up and top-down analyses. C1q was submitted to proteolytic digestion by a combination of collagenase and trypsin for bottom-up analyses. In addition to classical LC-MS/MS analyses, which provided reliable identification of hydroxylated proline and lysine residues, sugar loss-triggered MS(3) scans were acquired on an LTQ-Orbitrap (Linear Quadrupole Ion Trap Orbitrap) instrument to strengthen the localization of glucosyl-galactosyl disaccharide moieties on hydroxylysine residues. Top-down analyses performed on the same instrument allowed high accuracy and high resolution mass measurements of the intact full-length C1q polypeptide chains and the iterative fragmentation of the proteins in the MS(n) mode. This study illustrates the usefulness of combining the two complementary analytical approaches to obtain a detailed characterization of the post-translational modification pattern of the collagen like domain of C1q and highlights the structural heterogeneity of individual molecules. Most importantly, three lysine residues of the collagen-like domain, namely Lys(59) (A chain), Lys(61) (B chain), and Lys(58) (C chain), were unambiguously shown to be completely unmodified. These lysine residues are located about halfway along the collagen-like fibers. They are thus fully available and in an appropriate position to interact with the C1r and C1s protease partners of C1q and are therefore likely to play an essential role in C1 assembly. PMID- 20008837 TI - Soil respiration fluxes in a temperate mixed forest: seasonality and temperature sensitivities differ among microbial and root-rhizosphere respiration. AB - Although soil respiration, a major CO(2) flux in terrestrial ecosystems, is known to be highly variable with time, the response of its component fluxes to temperature and phenology is less clear. Therefore, we partitioned soil respiration (SR) into microbial (MR) and root-rhizosphere respiration (RR) using small root exclusion treatments in a mixed mountain forest in Switzerland. In addition, fine root respiration (FRR) was determined with measurements of excised roots. RR and FRR were strongly related to each other (R(2) = 0.92, n = 7), with RR contributing about 46% and FRR about 32% to total SR. RR rates increased more strongly with temperature (Q(10) = 3.2) than MR rates (Q(10) = 2.3). Since the contribution of RR to SR was found to be higher during growing (50%) than during dormant periods (40%), we separated the 2-year data set into phenophases. During the growing period of 2007, the temperature sensitivity of RR (Q(10) = 2.5, R(2) = 0.62) was similar to that of MR (Q(10) = 2.2, R(2) = 0.57). However, during the dormant period of 2006/2007, RR was not related to soil temperature (R(2) = 0.44, n.s.), in contrast to MR (Q(10) = 7.2; R(2) = 0.92). To better understand the influence of plant activity on root respiration, we related RR and FRR rates to photosynthetic active radiation (both R(2) = 0.67, n = 7, P = 0.025), suggesting increased root respiration rates during times with high photosynthesis. During foliage green-up in spring 2008, i.e., from bud break to full leaf expansion, RR increased by a factor of 5, while soil temperature increased only by about 5 degrees C, leading to an extraordinary high Q(10) of 10.6; meanwhile, the contribution of RR to SR increased from 29 to 47%. This clearly shows that root respiration and its apparent temperature sensitivity highly depend on plant phenology and thus on canopy assimilation and carbon allocation belowground. PMID- 20008835 TI - Quantitative proteomics analysis of chondrogenic differentiation of C3H10T1/2 mesenchymal stem cells by iTRAQ labeling coupled with on-line two-dimensional LC/MS/MS. AB - The chondrogenic potential of multipotent mesenchymal stem cells (MSCs) makes them a promising source for cell-based therapy of cartilage defects; however, the exact intracellular molecular mechanisms of chondrogenesis as well as self renewal of MSCs remain largely unknown. To gain more insight into the underlying molecular mechanisms, we applied isobaric tag for relative and absolute quantitation (iTRAQ) labeling coupled with on-line two-dimensional LC/MS/MS technology to identify proteins differentially expressed in an in vitro model for chondrogenesis: chondrogenic differentiation of C3H10T1/2 cells, a murine embryonic mesenchymal cell line, was induced by micromass culture and 100 ng/ml bone morphogenetic protein 2 treatment for 6 days. A total of 1756 proteins were identified with an average false discovery rate <0.21%. Linear regression analysis of the quantitative data gave strong correlation coefficients: 0.948 and 0.923 for two replicate two-dimensional LC/MS/MS analyses and 0.881, 0.869, and 0.927 for three independent iTRAQ experiments, respectively (p < 0.0001). Among 1753 quantified proteins, 100 were significantly altered (95% confidence interval), and six of them were further validated by Western blotting. Functional categorization revealed that the 17 up-regulated proteins mainly comprised hallmarks of mature chondrocytes and enzymes participating in cartilage extracellular matrix synthesis, whereas the 83 down-regulated were predominantly involved in energy metabolism, chromatin organization, transcription, mRNA processing, signaling transduction, and cytoskeleton; except for a number of well documented proteins, the majority of these altered proteins were novel for chondrogenesis. Finally, the biological roles of BTF3l4 and fibulin-5, two novel chondrogenesis-related proteins identified in the present study, were verified in the context of chondrogenic differentiation. These data will provide valuable clues for our better understanding of the underlying mechanisms that modulate these complex biological processes and assist in the application of MSCs in cell based therapy for cartilage regeneration. PMID- 20008838 TI - Storage behavior and changes in concentrations of abscisic acid and gibberellins during dormancy break and germination in seeds of Phellodendron amurense var. wilsonii (Rutaceae). AB - The medicinal Asian plant genus Phellodendron is known to contain several very important compounds that have biological action. The main purpose of this study was to determine whether seeds of Phellodendron amurense var. wilsonii can be stored and to characterize their dormancy. Seeds of this taxon stored at -20 and 80 degrees C and in liquid nitrogen retained their high germinability, indicating that they have orthodox storage behavior. Intact seeds from freshly collected fruits were dormant and required 12 weeks of cold stratification at 4 degrees C for complete germination. Scarifying the seed coat was partially effective in breaking seed dormancy. Exogenous gibberellins (GA(3), GA(4) and GA(4+7)) promoted germination of scarified seeds, GA(4) and GA(4+7) being more effective than GA(3). Fluridone, an abscisic acid (ABA) biosynthesis inhibitor, was efficient in breaking dormancy, but it was less effective than GA(4) or GA(4+7) alone. Paclobutrazol, a GA biosynthesis inhibitor, inhibited seed germination, and the inhibitory effect was reversed completely by GA(4) and by GA(4+7). ABA content of seeds subjected to cold stratification or to incubation at 35/10 degrees C, which enhanced seed germination, was reduced about four- to sixfold compared to that of fresh seeds. Higher concentrations of GA(3), GA(4) and GA(7) were detected in nondormant seeds and in seeds with an emerged radicle than in fresh seeds. Present results seem to indicate that dormancy in P. amurense var. wilsonii seeds is imposed partially by the seed coat and partially by high ABA content. ABA content decreased and GA(3), GA(4) and GA(7) content increased during germination. PMID- 20008839 TI - Overexpression of phospho-eIF4E is associated with survival through AKT pathway in non-small cell lung cancer. AB - PURPOSE: The eukaryotic translation initiation factor complex 4E (eIF4E) is downstream in the mammalian target of rapamycin (mTOR) pathway. This study explored expression of eIF4E and its relationship with the PTEN/AKT and RAS/MEK/ERK pathways in non-small cell lung carcinoma (NSCLC). EXPERIMENTAL DESIGN: The status of phosphorylated eIF4E (p-eIF4E), phosphorylated AKT (p-AKT), PTEN, phosphorylated tuberin (p-TSC2), phosphorylated mTOR (p-mTOR), phosphorylated S6 (p-S6), and phosphorylated Erk1/2 (p-Erk1/2) was studied using immunohistochemical analysis applied to a tissue microarray containing 300 NSCLCs. Staining results for each antibody were compared with clinical and pathologic features, and the relationship between staining results was explored. RESULTS: Overexpression of p-eIF4E, p-AKT, p-TSC2, p-mTOR, p-S6, and p-Erk1/2 in NSCLC was found in 39.9%, 78.8%, 5.1%, 46.7%, 27.1%, and 16.6% of tumors, respectively. The phenotype of p-eIF4E correlated positively with that of p-AKT, p-TSC2, and p-S6 (P < 0.001). Overall survival in NSCLC patients was significantly shorter in cases with overexpression of p-eIF4E and p-AKT alone and in combination (log-rank P < 0.001, each). Cases with underexpression of PTEN were limited (6.4%), and this phenotype did not correlate with any clinical variable. In cluster analysis, the p-AKT/p-mTOR/p-eIF4E/p-S6-positive group had significantly shorter survival compared with the survival of all cases (P < 0.001). Multivariate analysis showed that p-eIF4E overexpression is an independent prognostic factor for NSCLC (P = 0.004). CONCLUSIONS: This study shows that p-eIF4E expression in addition to p-AKT predicts poor prognosis in NSCLC. Moreover, the correlation between expression of p-eIF4E with p-AKT, as well as p-TSC2 and p-S6, indicates that eIF4E activation through the AKT pathway plays an important role in the progression of NSCLC. PMID- 20008840 TI - Hepatocyte growth factor reduces susceptibility to an irreversible epidermal growth factor receptor inhibitor in EGFR-T790M mutant lung cancer. AB - PURPOSE: The secondary T790M mutation in epidermal growth factor receptor (EGFR) is the most frequent cause of acquired resistance to the reversible EGFR tyrosine kinase inhibitors (EGFR-TKI), gefitinib and erlotinib, in lung cancer. Irreversible EGFR-TKIs are expected to overcome the reversible EGFR-TKI resistance of lung cancer harboring T790M mutation in EGFR. However, it is clear that resistance may also develop to this class of inhibitors. We showed previously that hepatocyte growth factor (HGF) induced gefitinib resistance of lung cancer harboring EGFR-activating mutations. Here, we investigated whether HGF induced resistance to the irreversible EGFR-TKI, CL-387,785, in lung cancer cells (H1975) harboring both L858R activating mutation and T790M secondary mutation in EGFR. EXPERIMENTAL DESIGN: CL-387,785 sensitivity and signal transduction in H1975 cells were examined in the presence or absence of HGF or HGF-producing fibroblasts with or without HGF-MET inhibitors. RESULTS: HGF reduced susceptibility to CL-387,785 in H1975 cells. Western blotting and small interfering RNA analyses indicated that HGF-induced hyposensitivity was mediated by the MET/phosphoinositide 3-kinase/Akt signaling pathway independent of EGFR, ErbB2, ErbB3, and ErbB4. Hyposensitivity of H1975 cells to CL-387,785 was also induced by coculture with high-level HGF-producing lung fibroblasts. The hyposensitivity was abrogated by treatment with anti-HGF neutralizing antibody, HGF antagonist NK4, or MET-TKI. CONCLUSIONS: We showed HGF-mediated hyposensitivity as a novel mechanism of resistance to irreversible EGFR-TKIs. It will be clinically valuable to investigate the involvement of HGF-MET-mediated signaling in de novo and acquired resistance to irreversible EGFR-TKIs in lung cancer harboring T790M mutation in EGFR. PMID- 20008841 TI - Prediction of survival following first-line chemotherapy in men with castration resistant metastatic prostate cancer. AB - PURPOSE: We sought to evaluate predictors of overall survival following progression after systemic chemotherapy in men with metastatic castration resistant prostate cancer. EXPERIMENTAL DESIGN: For our study population, we used the TAX327 multicenter randomized phase III trial comparing administration of docetaxel and prednisone every 3 weeks, weekly administration of docetaxel and prednisone, and administration of mitoxantrone and prednisone every 3 weeks. Progression was defined as the earliest of prostate-specific antigen (PSA), tumor, or pain progression. We analyzed predictors of postprogression survival according to both prechemotherapy and postchemotherapy variables with adjustment for potential confounders. RESULTS: Among 1,006 men, 640 had evaluable information on protocol-defined progression leading to further therapy. Median postprogression survival was 14.5 months. In the multivariable analysis, several pretreatment factors were associated with postprogression survival: pain, performance status, alkaline phosphatase, number of sites of metastatic disease, liver metastases, hemoglobin, PSA, and time since diagnosis. In addition, we found that the number of progression factors (PSA, pain, and tumor size), the duration of first-line chemotherapy, and whether progression occurred during chemotherapy independently predicted postprogression survival. We found evidence for the benefit of continuation of chemotherapy beyond progression only for men who had isolated worsening of pain. A nomogram was constructed and internally validated with a concordance index of 0.70. CONCLUSIONS: An internally validated model to predict postchemotherapy survival was developed. Evaluation of men in the postdocetaxel setting should consider the type of progression, duration of therapy, and known pretreatment prognostic factors. Definitions of progression in castration-resistant prostate cancer that include pain should also consider composite measures of tumor or PSA progression. External validation is planned. PMID- 20008842 TI - A high-throughput pharmaceutical screen identifies compounds with specific toxicity against BRCA2-deficient tumors. AB - PURPOSE: Hereditary breast cancer is partly explained by germline mutations in BRCA1 and BRCA2. Although patients carry heterozygous mutations, their tumors have typically lost the remaining wild-type allele. Selectively targeting BRCA deficiency may therefore constitute an important therapeutic approach. Clinical trials applying this principle are underway, but it is unknown whether the compounds tested are optimal. It is therefore important to identify alternative compounds that specifically target BRCA deficiency and to test new combination therapies to establish optimal treatment strategies. EXPERIMENTAL DESIGN: We did a high-throughput pharmaceutical screen on BRCA2-deficient mouse mammary tumor cells and isogenic controls with restored BRCA2 function. Subsequently, we validated positive hits in vitro and in vivo using mice carrying BRCA2-deficient mammary tumors. RESULTS: Three alkylators-chlorambucil, melphalan, and nimustine displayed strong and specific toxicity against BRCA2-deficient cells. In vivo, these showed heterogeneous but generally strong BRCA2-deficient antitumor activity, with melphalan and nimustine doing better than cisplatin and the poly (ADP-ribose)-polymerase inhibitor olaparib (AZD2281) in this small study. In vitro drug combination experiments showed synergistic interactions between the alkylators and olaparib. Tumor intervention studies combining nimustine and olaparib resulted in recurrence-free survival exceeding 330 days in 3 of 5 animals tested. CONCLUSIONS: We generated and validated a platform for identification of compounds with specific activity against BRCA2-deficient cells that translates well to the preclinical setting. Our data call for the re evaluation of alkylators, especially melphalan and nimustine, alone or in combination with the poly-(ADP-ribose)-polymerase inhibitors, for the treatment of breast cancers with a defective BRCA pathway. PMID- 20008843 TI - Quantified tumor t1 is a generic early-response imaging biomarker for chemotherapy reflecting cell viability. AB - PURPOSE: Identification of a generic response biomarker by comparison of chemotherapeutics with different action mechanisms on several noninvasive biomarkers in experimental tumor models. EXPERIMENTAL DESIGN: The spin-lattice relaxation time of water protons (T(1)) was quantified using an inversion recovery-TrueFISP magnetic resonance imaging method in eight different experimental tumor models before and after treatment at several different time points with five different chemotherapeutics. Effects on T(1) were compared with other minimally invasive biomarkers including vascular parameters, apparent diffusion coefficient, and interstitial fluid pressure, and were correlated with efficacy at the endpoint and histologic parameters. RESULTS: In all cases, successful chemotherapy significantly lowered tumor T(1) compared with vehicle and the fractional change in T(1) (DeltaT(1)) correlated with the eventual change in tumor size (range: r(2) = 0.21, P < 0.05 to r(2) = 0.73, P < 0.0001), except for models specifically resistant to that drug. In RIF-1 tumors, interstitial fluid pressure was decreased, but apparent diffusion coefficient and permeability increased in response to the microtubule stabilizer patupilone and 5 fluorouracil. Although DeltaT(1) was small (maximum of -20%), the variability was very low (5%) compared with other magnetic resonance imaging methods (24-48%). Analyses ex vivo showed unchanged necrosis, increased apoptosis, and decreased %Ki67 and total choline, but only Ki67 and choline correlated with DeltaT(1). Correlation of Ki67 and DeltaT(1) were observed in other models using patupilone, paclitaxel, a VEGF-R inhibitor, and the mammalian target of rapamycin inhibitor everolimus. CONCLUSIONS: These results suggest that a decrease in tumor T(1) reflects hypocellularity and is a generic marker of response. The speed and robustness of the method should facilitate its use in clinical trials. PMID- 20008844 TI - Inhibition of Vascular Endothelial Growth Factor Receptor-1/Wnt/{beta}-catenin Crosstalk Leads to Tumor Cell Death. AB - Two genes are considered synthetic lethal if mutation of either alone allows cell viability, whereas simultaneous mutation leads to cell death. A synthetic lethal screen unveiled the dependency of Wnt/beta-catenin-addicted colorectal cancer cells on vascular endothelial growth factor receptor-1 kinase activity and suggested a novel therapeutic approach for this malignancy. (Clin Cancer Res 2009;15(24):7453-5). PMID- 20008846 TI - From the Editor. PMID- 20008845 TI - Hyaluronan-CD44 Interactions in Cancer: Paradoxes and Possibilities. AB - Hyaluronan is a prominent component of the micro-environment in most malignant tumors and can be prognostic for tumor progression. Extensive experimental evidence in animal models implicates hyaluronan interactions in tumor growth and metastasis, but it is also evident that a balance of synthesis and turnover by hyaluronidases is critical. CD44, a major hyaluronan receptor, is commonly but not uniformly associated with malignancy, and is frequently used as a marker for cancer stem cells in human carcinomas. Multivalent interactions of hyaluronan with CD44 collaborate in driving numerous tumor-promoting signaling pathways and transporter activities. It is widely accepted that hyaluronan-CD44 interactions are crucial in both malignancy and resistance to therapy, but major challenges for future research in the field are the mechanism of activation of hyaluronan CD44 signaling in cancer cells, the relative importance of variant forms of CD44 and other hyaluronan receptors, e.g., Rhamm, in different tumor contexts, and the role of stromal versus tumor cell production and turnover of hyaluronan. Despite these caveats, it is clear that hyaluronan-CD44 interactions are an important target for translation into the clinic. Among the approaches that show promise are antibodies and vaccines to specific variants of CD44 that are uniquely expressed at critical stages of progression of a particular cancer, hyaluronidase mediated reduction of barriers to drug access, and small hyaluronan oligosaccharides that attenuate constitutive hyaluronan-receptor signaling and enhance chemosensitivity. In addition, hyaluronan is being used to tag drugs and delivery vehicles for targeting of anticancer agents to CD44-expressing tumor cells. (Clin Cancer Res 2009;15(24):7462-8). PMID- 20008847 TI - Resistance to Targeted Therapies: Refining Anticancer Therapy in the Era of Molecular Oncology. AB - The advent of targeted therapy for treatment of human cancers has added significantly to our armamentarium as we strive to prolong patient survival while minimizing toxicity. In cancers driven by a dominant oncogene, targeted therapies have led to remarkable improvements in response and survival, whereas in others the outcome has been more modest. One key aspect toward realizing the potential of targeted therapies is a better understanding of the intrinsic or acquired resistance mechanisms that limit their efficacy. The articles in this CCR Focus provide insights into molecular mechanisms of resistance to targeted therapy. Recent discoveries of the molecular pathways that mediate intrinsic resistance to targeted therapy have led to the identification of predictive biomarkers that allow for better patient selection for front line treatment. Equally important, the identification of mechanisms of acquired resistance following front line therapy has led to the discovery of novel agents that overcome these resistance mechanisms. Improving the efficacy of targeted therapies in the future will require expanding our understanding of resistance mechanisms, the development of new generations of rationally designed targeted agents, and translating this information to the clinic to select patients for appropriate therapy. (Clin Cancer Res 2009;15(24):7471-8). PMID- 20008849 TI - Biomarkers of Resistance to Epidermal Growth Factor Receptor Monoclonal Antibodies in Patients with Metastatic Colorectal Cancer. AB - Genomics and proteomics have held out the promise of individualized medicine for the last 10 or 20 years, but clinical medicine has not yet delivered on this promise. Some cancers, such as breast cancer and some hematologic malignancies, have been at the forefront of individualized therapeutic approaches by integrating molecular biomarkers into treatment decision algorithms. Until recently, the treatment of colorectal cancer (CRC) has lagged behind these other cancers in this regard and therapeutic decisions have been solely empirical. Data from various clinical trials and translational studies have now opened the door for individualized treatment approaches by identifying patients with metastatic CRC who are most likely to benefit from antibodies against the epidermal growth factor receptor (EGFR), cetuximab and panitumumab. Activating mutations of KRAS, a downstream mediator of EGFR signaling, has been shown to render EGFR antibodies ineffective, such that analyzing tumors for these mutations has become mandatory before the use of EGFR antibodies is considered in CRC. Beyond KRAS, several additional biomarkers are currently being investigated as potential positive or negative predictors for the efficacy of EGFR-targeted therapy. Most of these markers are alterations of molecules integrated in the EGFR pathway. This review will focus on the type and quality of evidence that has been gathered to date to predict resistance to monoclonal antibodies against EGFR in CRC. (Clin Cancer Res 2009;15(24):7492-501). PMID- 20008850 TI - Resistance to Epidermal Growth Factor Receptor Tyrosine Kinase Inhibitors in Non Small Cell Lung Cancer. AB - Gefitinib and erlotinib are ATP competitive inhibitors of the epidermal growth factor receptor (EGFR) tyrosine kinase and are approved around the world for the treatment of patients with non-small cell lung cancer (NSCLC). Somatic mutations in the EGFR are found in 10 to 40% of patients with NSCLC. Patients with sensitizing somatic mutations of EGFR treated with gefitinib or erlotinib have an initial clinical response of 60 to 80%, approximately twice as high as the responses associated with the administration of conventional platinum-based chemotherapy. However, the efficacy of EGFR tyrosine kinase inhibitors (TKI) is limited by either primary (de novo) or acquired resistance after therapy and investigations to define the mechanisms of resistance are active areas of ongoing preclinical and clinical studies. Primary resistance is typically caused by other somatic mutations in genes such as KRAS, which also have an impact on the EGFR signaling pathway or by mutations in the EGFR gene that are not associated with sensitivity to EGFR-TKIs. Two established mechanisms of acquired resistance are caused by additional mutations in the EGFR gene acquired during the course of treatment that change the protein-coding sequence or by amplification of another oncogene signaling pathway driven by the MET oncogene. This review focuses on characterized mechanisms of resistance to the EGFR TKIs and efforts to overcome the problem of resistance aimed at improving the therapy of patients with NSCLC. (Clin Cancer Res 2009;15(24):7502-9). PMID- 20008848 TI - Resistance to Trastuzumab in Breast Cancer. AB - HER2 is a transmembrane oncoprotein encoded by the HER2/neu gene and is overexpressed in approximately 20 to 25% of invasive breast cancers. It can be therapeutically targeted by trastuzumab, a humanized IgG1 kappa light chain monoclonal antibody. Although trastuzumab is currently considered one of the most effective treatments in oncology, a significant number of patients with HER2 overexpressing breast cancer do not benefit from it. Understanding the mechanisms of action and resistance to trastuzumab is therefore crucial for the development of new therapeutic strategies. This review discusses proposed trastuzumab mode of action as well as proposed mechanisms for resistance. Mechanisms for resistance are grouped into four main categories: (1) obstacles preventing trastuzumab binding to HER2; (2) upregulation of HER2 downstream signaling pathways; (3) signaling through alternate pathways; and (4) failure to trigger an immune mediated mechanism to destroy tumor cells. These potential mechanisms through which trastuzumab resistance may arise have been used as a guide to develop drugs, presently in clinical trials, to overcome resistance. The mechanisms conferring trastuzumab resistance, when completely understood, will provide insight on how best to treat HER2-overexpressing breast cancer. The understanding of each mechanism of resistance is therefore critical for the educated development of strategies to overcome it, as well as for the development of tools that would allow definitive and efficient patient selection for each therapy. (Clin Cancer Res 2009;15(24):7479-91). PMID- 20008851 TI - Resistance to Tyrosine Kinase Inhibitors in Gastrointestinal Stromal Tumors. AB - Gastrointestinal stromal tumors (GIST) are the most common type of sarcoma in the gastrointestinal tract. Surgery is the primary treatment modality, but many patients suffer disease recurrence or metastasis. Fortunately, the management of advanced GIST has been revolutionized by the use of small molecule kinase inhibitors that target the underlying pathogenetic mutant kinases found in the vast majority of cases. Approximately 85% of GISTs have oncogenic mutations in KIT, allowing for constitutive kinase activation that is responsible for cellular proliferation and survival. About 5 to 7% of GISTs have activating mutations of the homologous platelet-derived growth factor receptor alpha (PDGFRA) kinase. The progression-free and overall survival of patients with advanced disease is greatly improved by treatment with the kinase inhibitors imatinib and sunitinib. However, the emergence of drug-resistant tumor clones limits the long-term benefit of these drugs in most patients. Resistance to these kinase inhibitors is associated with distinctive clinical and molecular features, with the development of secondary mutations of the oncogenic kinase being the most common mechanism. We review the molecular basis of GIST response and/or resistance to TKIs, and discuss strategies to prevent and/or overcome drug resistance. These concepts are directly relevant to the development of targeted molecular therapy for other solid tumors. (Clin Cancer Res 2009;15(24):7510-8). PMID- 20008852 TI - Mechanisms of Resistance to Imatinib and Second-Generation Tyrosine Inhibitors in Chronic Myeloid Leukemia. AB - Targeted therapy in the form of selective tyrosine kinase inhibitors (TKI) has transformed the approach to management of chronic myeloid leukemia (CML) and dramatically improved patient outcome to the extent that imatinib is currently accepted as the first-line agent for nearly all patients presenting with CML, regardless of the phase of the disease. Impressive clinical responses are obtained in the majority of patients in chronic phase; however, not all patients experience an optimal response to imatinib, and furthermore, the clinical response in a number of patients will not be sustained. The process by which the leukemic cells prove resistant to TKIs and the restoration of BCR-ABL1 signal transduction from previous inhibition has initiated the pursuit for the causal mechanisms of resistance and strategies by which to surmount resistance to therapeutic intervention. ABL kinase domain mutations have been extensively implicated in the pathogenesis of TKI resistance, however, it is increasingly evident that the presence of mutations does not explain all cases of resistance and does not account for the failure of TKIs to eliminate minimal residual disease in patients who respond optimally. The focus of exploring TKI resistance has expanded to include the mechanism by which the drug is delivered to its target and the impact of drug influx and efflux proteins on TKI bioavailability. The limitations of imatinib have inspired the development of second generation TKIs in order to overcome the effect of resistance to this primary therapy. (Clin Cancer Res 2009;15(24):7519-27). PMID- 20008853 TI - Vascular Endothelial Growth Factor Receptor-1 Is Synthetic Lethal to Aberrant {beta}-Catenin Activation in Colon Cancer. AB - PURPOSE: The Wnt/beta-catenin (beta-cat) signaling cascade is a key regulator of development, and dysregulation of Wnt/beta-cat contributes to selected cancers, such as colorectal, breast, and hepatocellular carcinoma, through abnormal activation of Wnt target genes. To identify novel modulators of the Wnt/beta-cat pathway that may emerge as therapeutic targets, we did an unbiased high throughput RNA interference screen. EXPERIMENTAL DESIGN: A synthetic oligonucleotide small interfering RNA library targeting 691 known and predicted human kinases was screened in Wnt3a-stimulated human cells in a live cell luciferase assay for modulation of Wnt/beta-cat-dependent transcription. Follow up studies of a selected high-confidence "hit" were conducted. RESULTS: A robust quartile-based statistical analysis and secondary screen yielded several kinases worthy of further investigation, including Cdc2L1, Lmtk3, Pank2, ErbB3, and, of note, vascular endothelial growth factor receptor (VEGFR)1/Flt1, a receptor tyrosine kinase (TK) with putative weak kinase activity conventionally believed to be a negative regulator of angiogenesis. A series of loss-of-function, genetic null, and VEGFR TK inhibitor assays further revealed that VEGFR1 is a positive regulator of Wnt signaling that functions in a glycogen synthase kinase-3beta (GSK3beta)-independent manner as a potential synthetic lethal target in Wnt/beta cat-addicted colon carcinoma cells. CONCLUSIONS: This unanticipated non endothelial link between VEGFR1 TK activity and Wnt/beta-cat signaling may refine our understanding of aberrant Wnt signaling in colon carcinoma and points to new combinatorial therapeutics targeted to the tumor cell compartment, rather than angiogenesis, in the context of colon cancer. (Clin Cancer Res 2009;15(24):7529 37). PMID- 20008854 TI - Novel Inhibitors of Fatty Acid Synthase with Anticancer Activity. AB - PURPOSE: Fatty acid synthase (FASN) is overexpressed in human breast carcinoma. The natural polyphenol (-)-epigallocatechin-3-gallate blocks in vitro FASN activity and leads to apoptosis in breast cancer cells without any effects on carnitine palmitoyltransferase-1 (CPT-1) activity, and in vivo, does not decrease body weight. We synthesized a panel of new polyphenolic compounds and tested their effects on breast cancer models. EXPERIMENTAL DESIGN: We evaluated the in vitro effects of the compounds on breast cancer cell growth (SK-Br3, MCF-7, and MDA-MB-231), apoptosis [as assessed by cleavage of poly(ADP-ribose) polymerase], cell signaling (HER2, ERK1/2, and AKT), and fatty acid metabolism enzymes (FASN and CPT-1). In vivo, we have evaluated their antitumor activity and their effect on body weight in a mice model of BT474 breast cancer cells. RESULTS: Two compounds potently inhibited FASN activity and showed high cytotoxicity. Moreover, the compounds induced apoptosis and caused a marked decrease in the active forms of HER2, AKT, and ERK1/2 proteins. Interestingly, the compounds did not stimulate CPT-1 activity in vitro. We show evidence that one of the FASN inhibitors blocked the growth of BT474 breast cancer xenografts and did not induce weight loss in vivo. CONCLUSIONS: The synthesized polyphenolic compounds represent a novel class of FASN inhibitors, with in vitro and in vivo anticancer activity, that do not exhibit cross-activation of beta-oxidation and do not induce weight loss in animals. One of the compounds blocked the growth of breast cancer xenografts. These FASN inhibitors may represent new agents for breast cancer treatment. (Clin Cancer Res 2009;15(24):7608-15). PMID- 20008855 TI - A Phase I Clinical Trial of CHT-25 a 131I-Labeled Chimeric Anti-CD25 Antibody Showing Efficacy in Patients with Refractory Lymphoma. AB - PURPOSE: There is a need for new treatments for Hodgkin and T-cell lymphoma due to the development of drug resistance in a proportion of patients. This phase I study of radioimmunotherapy used CHT-25, a chimeric antibody to the alpha-chain of the interleukin-2 receptor, CD25, conjugated to iodine-131 ((131)I) in patients with refractory CD25-positive lymphomas. EXPERIMENTAL DESIGN: Fifteen patients were treated (Hodgkin lymphoma, 12; angioimmunoblastic T-cell lymphoma, 1; adult T-cell leukemia/lymphoma, 2). Tumor was monitored by computed tomography and in all but two patients by (18)F-fluorodeoxyglucose positron emission tomography. RESULTS: There were no grade 3 or 4 infusion reactions. At the maximum tolerated dose of 1,200 MBq/m(2), the major side effect was delayed myelotoxicity with the nadir for platelets at 38 days and for neutrophils at 53 days. One patient treated with 2,960 MBq/m(2) developed prolonged grade 4 neutropenia and thrombocytopenia and died of Pneumocystis jiroveci pneumonia. Nonhematologic toxicity was mild. Single photon emission computer tomography imaging showed tumor-specific uptake and retention of (131)I and no excessive retention in normal organs. Of nine patients receiving >/=1,200 MBq/m(2), six responded (three complete response and three partial response); one of six patients with administered radioactivity of or =45 years). Hearing impairment was defined as a pure-tone average of thresholds at 0.5, 1, 2, and 4 kHz greater than 25-dB HL [hearing level]. Descriptive analysis, generalized additive models, and alternating logistic regression models were used to examine the birth cohort effect. Controlling for age, with every 5-year increase in birth year, the odds of having hearing impairment were 13% lower in men (odds ratio = 0.87, 95% confidence interval: 0.83, 0.92) and 6% lower in women (odds ratio = 0.94, 95% confidence interval: 0.89, 0.98). These results suggest that 1) older adults may be retaining good hearing longer than previous generations and 2) modifiable factors contribute to hearing impairment in adults. PMID- 20008890 TI - c-MYC depletion potentiates cisplatin-induced apoptosis in head and neck squamous cell carcinoma: involvement of TSP-1 up-regulation. PMID- 20008891 TI - Blue-light hazard from CO2 arc welding of mild steel. AB - OBJECTIVES: The objective was to quantify the blue-light hazard from CO(2) arc welding of mild steel. METHODS: The spectral radiance of arcs in CO(2) arc welding of mild steel was measured for solid and flux-cored wires at welding currents of 120-480 A. Effective blue-light radiance and the maximum acceptable exposure duration were calculated from the spectral radiance using their definitions in American Conference of Governmental Industrial Hygienists guidelines. RESULTS: The effective blue-light radiance ranged from 22.9 to 213.1 Wcm(-2)sr(-1). The corresponding maximum acceptable exposure duration was only 0.47-4.36 s, meaning that the total daily exposure to the welding arc without eye protection should not exceed this duration. CONCLUSIONS: It is very hazardous to view the arcs in CO(2) arc welding of mild steel. Welders and their helpers should use appropriate eye protectors in these arc-welding operations. Also, they should avoid direct light exposure when starting an arc-welding operation. PMID- 20008892 TI - Managing dermal risk: moving on from gloves. PMID- 20008893 TI - Hydration, hydration, hydration. AB - Throughout the world, large numbers of manual workers perform physically demanding labour in conditions of high environmental heat stress. Although the importance of adequate hydration in combating heat stress is universally recognized, studies in a range of worker groups have demonstrated a disturbingly poor hydration level in a high proportion of at-risk workers. Management of work in hot environments traditionally focuses on environmental monitoring, while strategies to promote and ensure good hydration behaviour are often haphazard at best. An example is given of simple guidelines for adequate and appropriate fluid intake and practical recommendations to foster compliance. PMID- 20008894 TI - Comparing location memory for 4 sensory modalities. AB - Stimuli from all sensory modalities can be linked to places and thus might serve as navigation cues. We compared performance for 4 sensory modalities in a location memory task: Black-and-white drawings of free forms (vision), 1-s manipulated environmental sounds (audition), surface textures of natural and artificial materials (touch), and unfamiliar smells (olfaction) were presented in 10 cubes. In the learning stage, participants walked to a cube, opened it, and perceived its content. Subsequently, in a relocation task, they placed each stimulus back in its original location. Although the proportion of correct locations selected just failed to yield significant differences between the modalities, the proportion of stimuli placed in the vicinity of the correct location or on the correct side of the room was significantly higher for vision than for touch, olfaction, and audition. These outcomes suggest that approximate location memory is superior for vision compared with other sensory modalities. PMID- 20008895 TI - Photosynthesis in lightfleck areas of homobaric and heterobaric leaves. AB - Leaves within a canopy are exposed to a spatially and temporally fluctuating light environment which may cause lateral gradients in leaf internal CO(2) concentration and diffusion between shaded and illuminated areas. In previous studies it was hypothesized that lateral CO(2) diffusion may support leaf photosynthesis, but the magnitude of this effect is still not well understood. In the present study homobaric leaves of Vicia faba or heterobaric leaves of Glycine max were illuminated with lightflecks of different sizes, mimicking sunflecks. Photosynthetic properties of the lightfleck areas were assessed with combined gas exchange measurements and chlorophyll fluorescence imaging. Lateral diffusion in homobaric leaves with an interconnected intercellular air space stimulated photosynthesis and the effect was largest in small lightfleck areas, in particular when plants were under drought stress. Such effects were not observed in the heterobaric leaves with strongly compartmented intercellular gas spaces. It is concluded that lateral diffusion may significantly contribute to photosynthesis of lightfleck areas of homobaric leaves depending on lightfleck size, lateral diffusivity, and stomatal conductance. Since homobaric leaf structures have been reported for many plant species, it is hypothesized that leaf homobary may have an impact on overall plant performance under conditions with a highly heterogeneous light environment. PMID- 20008896 TI - Esophageal cyst producing CA19-9 and CA125. AB - The patient was a 59-year-old woman in whom computed tomography revealed a posterior mediastinal cyst and ovarian cystoma at a medical check-up in March 2007. Blood tests showed high CA19-9 and CA125 levels. She underwent left adnexectomy for ovarian cystoma in July 2008 and histopathological examination led to a diagnosis of dermoid cyst. The postoperative levels of CA19-9 and CA125 remained high. She developed dysphagia in February 2009, and the posterior mediastinal cyst showed a tendency to enlarge. Therefore, she underwent tumorectomy through a small thoracotomy. The cyst contained greenish fluid with CA19-9 and CA125 contents of 65,000 and 78,000 U/ml, respectively. Histologically, the cyst had a thickened wall, which contained two muscle layers, and was lined by squamous and pseudostratified ciliated epithelium. No cartilage or bronchial glands were identified. These findings led to a diagnosis of esophageal cyst. On immunohistochemical staining, the cyst-lining epithelial cells were positive for CA19-9 and CA125. The serum CA19-9 and CA125 levels returned to normal two months after surgery. We report a resected case of esophageal cyst producing CA19-9 and CA125. PMID- 20008897 TI - Impact of meteorological conditions on the occurrence of acute type A aortic dissections. AB - The impact of meteorological conditions on the occurrence of various cardiovascular events has been reported. The aim of this work was to study the correlations between weather conditions and the occurrence of type A acute aortic dissections (AADs). Between 1997 and 2007, all the medical records of patients who underwent surgery for type A AADs in Toulouse University Hospital (France) were reviewed. The clinical data were confronted with the meteorological data provided by the French national meteorological office (MeteoFrance) over the same period. Two hundred and six patients with spontaneous type A AADs underwent surgery during this period. The incidence of aortic dissection was higher in winter time than in summer (P=0.018). The days with aortic dissections were colder than those without aortic dissections (P=0.017). Statistical analysis highlighted a decrease of atmospheric temperature during the three days preceding the upset of the symptoms (P=0.0009). This work demonstrates a correlation between spontaneous type A AADs and low atmospheric temperature. PMID- 20008898 TI - Cardiac troponin I levels after cardiac surgery as predictor for in-hospital mortality. AB - PURPOSE: Troponin is a specific marker of myocardial damage. Increased troponins, however, are observed after almost all cardiac surgery. The clinical significance of this elevation is controversial. The aim of this study was to evaluate if troponin I (cTnI) measured 1 h after cardiac surgery provides additional information to identify patients at risk for hospital mortality. METHODS: Nine hundred and thirty-eight patients undergoing cardiac surgery between October 2006 and June 2008 served as development set. This group included 688 isolated CABGs and 250 valvular (+CABG) operations, and cTnI levels were measured 1 h (cTnI) after surgery. Hospital mortality, defined as death occurring at the Radboud University Nijmegen Medical Centre (UMCN) at any time after surgery, is the studied outcome. To assess the value of cTnI as a predictor for hospital mortality, receiver-operator characteristic (ROC) curves were used. The Youden index was used for identifying the best cut-off point. Five hundred and seventy nine patients undergoing cardiac surgery between July 2008 and February 2009 served as validation set. RESULTS: The median cTnI level was 1.3 microg/l, 75% inter-quartile range (IQR) 0.68-2.59 microg/l. Ten patients (1.1%) died, cTnI release of the dead, median: 6.8 microg/l was significantly higher than the measured values in the group of survivors, median: 1.3 microg/l (P<0.001). Regression analysis showed a significant correlation between cTnI and hospital mortality (P<0.001). The ROC indicates a cTnI level of 4.25 microg/l with a ROC of 0.80 as optimal cut-off point for predicting hospital mortality, with a sensitivity of 70% and a specificity of 89%. Addition of type of surgery, isolated CABG vs. valve surgery, acute vs. elective surgery and EuroSCORE class did not improve the ROCs. In the validation set, the median cTnI level was 1.17 microg/l. Fifty-six patients had a cTnI level >4.25 microg/l. Of the 579 patients, 11 patients (1.8%) died, six of them had a cTnI level >4.25 microg/l. CONCLUSION: Postoperative cTnI level, measured within the first hour after cardiac surgery, can identify a subgroup of patients with increased risk for hospital mortality. These patients may benefit from better monitoring, eventually with specific diagnostic and therapeutic interventions. PMID- 20008900 TI - Effects of dyskeratosis congenita mutations in dyskerin, NHP2 and NOP10 on assembly of H/ACA pre-RNPs. AB - Dyskeratosis congenita (DC) is a rare genetic syndrome that gives rise to a variety of disorders in affected individuals. Remarkably, all causative gene mutations identified to date share a link to telomere/telomerase biology. We found that the most prevalent dyskerin mutation in DC (A353V) did not affect formation of the NAF1-dyskerin-NOP10-NHP2 tetramer that normally assembles with nascent H/ACA RNAs in vivo. However, the A353V mutation slightly reduced pre-RNP assembly with the H/ACA-like domain of human telomerase RNA (hTR). In contrast, NHP2 mutations V126M and Y139H impaired association with NOP10, leading to major pre-RNP assembly defects with all H/ACA RNAs tested, including the H/ACA domain of hTR. Mutation R34W in NOP10 caused no apparent defect in protein tetramer formation, but it severely affected pre-RNP assembly with the H/ACA domain of hTR and a subset of H/ACA RNAs. Surprisingly, H/ACA sno/scaRNAs that encode miRNAs were not affected by the mutation R34W, and they were able to form pre-RNPs with NOP10-R34W. This indicates structural differences between H/ACA RNPs that encode miRNAs and those that do not. Altogether, our results suggest that, in addition to major defects in the telomere/telomerase pathways, some of the disorders occurring in DC may be caused by alteration of most H/ACA RNPs, or by only a subset of them. PMID- 20008901 TI - Dismutase-competent SOD1 mutant accumulation in myelinating Schwann cells is not detrimental to normal or transgenic ALS model mice. AB - Mutant superoxide dismutase 1 (SOD1) action within non-neuronal cells is implicated in damage to spinal motor neurons in a genetic form of amyotrophic lateral sclerosis (ALS). Central nervous system glial cells such as astrocytes and microglia drive progression in transgenic mutant SOD1 mice, however, the role of myelinating glia remains unclear. Specifically, peripheral myelinating glial cells are likely candidates for mediating degeneration of distal synapses and axons of motor neurons in ALS. Here, we examine the potential contribution of peripheral axon ensheathing Schwann cells to ALS by constructing transgenic mice expressing dismutase active mutant SOD1(G93A) driven by the myelin protein zero (P0) promoter. In this model, mutant SOD1 accumulation in Schwann cells was comparable to levels in mice ubiquitously expressing a SOD1(G93A) transgene that become paralysed. Growth, locomotion and survival of these P0-SOD1(G93A) mice were indistinguishable from normal animals. There was no evidence for spinal motor neuron loss, distal axonal degeneration and p75 neurotrophin receptor (p75(NTR)) upregulation in the periphery of P0-SOD1(G93A) mice, unlike transgenic SOD1(G93A) mice with presymptomatic p75(NTR) induction and death-signalling. Furthermore, Schwann cells were resistant to mutant SOD1 aggregation in vivo and in transfected primary cultures. Increasing mutant SOD1 synthesis in Schwann cells by cross-breeding transgenic P0-SOD1(G93A) and SOD1(G93A) mice did not affect disease onset or survival. We conclude that dismutase-competent mutant SOD1 accumulation within Schwann cells is not pathological to spinal motor neurons or deleterious to disease course in transgenic ALS model mice, in contrast to astrocytes and microglia. PMID- 20008902 TI - Screening for antiviral activities of isolated compounds from essential oils. AB - Essential oil of star anise as well as phenylpropanoids and sesquiterpenes, for example, trans-anethole, eugenol, beta-eudesmol, farnesol, beta-caryophyllene and beta-caryophyllene oxide, which are present in many essential oils, were examined for their antiviral activity against herpes simplex virus type 1 (HSV-1) in vitro. Antiviral activity was analyzed by plaque reduction assays and mode of antiviral action was determined by addition of the drugs to uninfected cells, to the virus prior to infection or to herpesvirus-infected cells. Star anise oil reduced viral infectivity by >99%, phenylpropanoids inhibited HSV infectivity by about 60-80% and sesquiterpenes suppressed herpes virus infection by 40-98%. Both, star anise essential oil and all isolated compounds exhibited anti-HSV-1 activity by direct inactivation of free virus particles in viral suspension assays. All tested drugs interacted in a dose-dependent manner with herpesvirus particles, thereby inactivating viral infectivity. Star anise oil, rich in trans anethole, revealed a high selectivity index of 160 against HSV, whereas among the isolated compounds only beta-caryophyllene displayed a high selectivity index of 140. The presence of beta-caryophyllene in many essential oils might contribute strongly to their antiviral ability. These results indicate that phenylpropanoids and sesquiterpenes present in essential oils contribute to their antiviral activity against HSV. PMID- 20008903 TI - Insulin-Mimetic Action of Rhoifolin and Cosmosiin Isolated from Citrus grandis (L.) Osbeck Leaves: Enhanced Adiponectin Secretion and Insulin Receptor Phosphorylation in 3T3-L1 Cells. AB - Citrus grandis (L.) Osbeck (red wendun) leaves have been used in traditional Chinese medicine to treat several illnesses including diabetes. However, there is no scientific evidence supporting these actions and its active compounds. Two flavone glycosides, rhoifolin and cosmosiin were isolated for the first time from red wendun leaves and, identified these leaves are rich source for rhoifolin (1.1%, w/w). In differentiated 3T3-L1 adipocytes, rhoifolin and cosmosiin showed dose-dependent response in concentration range of o.oo1-5 MUM and 1-20 MUM, respectively, in biological studies beneficial to diabetes. Particularly, rhoifolin and cosmosiin at 0.5 and 20 MUM, respectively showed nearly similar response to that 10 nM of insulin, on adiponectin secretion level. Furthermore, 5 MUM of rhoifolin and 20 MUM of cosmosiin showed equal potential with 10 nM of insulin to increase the phosphorylation of insulin receptor-beta, in addition to their positive effect on GLUT4 translocation. These findings indicate that rhoifolin and cosmosiin from red wendun leaves may be beneficial for diabetic complications through their enhanced adiponectin secretion, tyrosine phosphorylation of insulin receptor-beta and GLUT4 translocation. PMID- 20008904 TI - Exploring the determinants of unsafe abortion: improving the evidence base in Mexico. AB - BACKGROUND: Despite the realized importance of unsafe abortion as a global health problem, reliable data are difficult to obtain, especially in countries where abortion is illegal. Estimates for most developing countries are based on limited and incomplete sources of data. In Mexico, studies have been undertaken to improve estimates of induced abortion but the determinants of unsafe abortion have not been explored. METHODS: We analysed data from the 2006 Mexican National Demographic Survey. The sample comprises 14 859 reported pregnancies in women between 15 and 55 years old, of which 966 report having had an abortion in the 5 years preceding the survey. We use logistic regression to explore the relationship between unsafe abortion and various socio-economic and demographic characteristics. FINDINGS: We estimate that 44% of abortions have been induced and 16.5% of those were unsafe. We find three variables to be positively and significantly associated with the probability of having an induced abortion: (1) whether the woman reported that the pregnancy was mistimed (OR = 4.5, 95% CI = 1.95-10.95); (2) whether the woman reported that the pregnancy was unwanted (OR = 2.86, 95% CI = -1.40-5.88); and (3) if the woman had three or more children at the time of the abortion (OR = 3.73, 95% CI = 1.20-11.65). There is a steep socio economic gradient in the probability of having an unsafe abortion: poorer women are more likely to have an unsafe abortion than richer women (OR = 2.48, 95% CI = 1.09-5.63); women with 6-9 years of education (OR = 0.30, 95% CI = 0.11-0.81) and with more than 13 years of education are less likely to have an unsafe abortion (OR = 0.065, 95% CI = 0.01-0.43), and women with indigenous origin are more likely to have an unsafe abortion (OR = 5.44, 95% CI = 1.91-15.51). Thus, the probability for poor women with less than 5 years of education and indigenous origin is nine times higher compared with rich, educated and not indigenous women. We also find marked geographical inequities as women living in the poorest states have a higher risk of having an unsafe abortion. INTERPRETATION: This analysis has explored the determinants of unsafe abortion and has demonstrated that there are large socio-economic and geographical inequities in unsafe abortions in Mexico. Further efforts are required to improve the measurement and monitoring of trends in unsafe abortions in developing countries. PMID- 20008906 TI - A survey of attitudes towards methicillin-resistant Staphylococcus aureus bacteraemias amongst United Kingdom microbiologists. AB - OBJECTIVES: Methicillin-resistant Staphylococcus aureus (MRSA) bloodstream infections (BSIs) in the UK are common and associated with significant morbidity and mortality. Vancomycin is the usual first-line therapy. However, vancomycin treatment of BSIs due to MRSA strains with vancomycin MICs of 1-2 mg/L is successful in <10% of cases. No consensus exists on when to use newer agents, particularly when vancomycin MICs are >1 mg/L. We therefore surveyed UK practices of the management of MRSA BSIs due to isolates with increased vancomycin MICs. METHODS: Five hundred and seventy-one UK consultant microbiologists were contacted via e-mail and asked to take part in an online survey, hosted at www.surveymonkey.com. Responses were collated by the website, downloaded and analysed in a Microsoft Excel (Microsoft Corporation) spreadsheet. RESULTS: One hundred and eight respondents participated in the survey. Only 32.7% routinely measure MICs, mostly by Etest. Forty-two percent use vancomycin alone for removable-focus infections, whilst for infections of cardiac or orthopaedic origin, 49% would add rifampicin. Few respondents use daptomycin, linezolid or tigecycline empirically. Sixty-nine percent would use linezolid as a second-line agent, with only 19% opting for daptomycin. For an isolate with a vancomycin MIC of 4 mg/L, respondents would use daptomycin (81%) or linezolid (91%) in patients with a poor clinical response. CONCLUSIONS: Vancomycin is the mainstay therapy for MRSA BSIs, even when MICs are not measured or raised, despite evidence of high failure rates. The use of newer agents frequently does not follow European or US licensed indications, may be inappropriate and may result in avoidable deaths. PMID- 20008905 TI - Low-abundance HIV species and their impact on mutational profiles in patients with virological failure on once-daily abacavir/lamivudine/zidovudine and tenofovir. AB - BACKGROUND: HIV clonal genotypic analysis (CG) was used to investigate whether a more sensitive analysis method would detect additional low-abundance mutations compared with population genotyping (PG) in antiretroviral-naive patients who experienced virological failure (VF) during treatment with abacavir/lamivudine/zidovudine and tenofovir. METHODS: HIV was analysed by PG and CG (771 baseline and 657 VF clones) from subjects with VF (confirmed HIV RNA > or = 400 copies/mL at 24-48 weeks). RESULTS: Fourteen of 123 subjects (11%) met VF criteria; their median baseline HIV RNA was 5.4 log(10) copies/mL, and 4.0 log(10) copies/mL at VF. By baseline PG, 2/14 had HIV-1 with nucleoside reverse transcriptase inhibitor (NRTI) or non-NRTI mutations. By baseline CG, 9/14 had HIV-1 with NNRTI and/or NRTI mutations; 7/9 had study drug-associated mutations. By PG at VF, 10/14 had selected for resistance mutations [2, K65R; 1, M184V; and 7, thymidine analogue mutations (TAMs) +/- M184V]. By CG at VF, for subjects with TAMs, T215F was more commonly detected (5/14 samples) than T215Y (2/14). For one subject who selected K65R at VF, both K65R-containing clones and TAM-containing clones (both T215A and T215F) were observed independently but not conjunctively in the same clone in a post-VF sample. CONCLUSIONS: The majority of subjects with VF had major and minor mutations detected at VF; CG detected additional low abundance variants at baseline and VF that could have influenced mutation selection pathways. Both PG and CG data suggest TAMs, not K65R selection, are the preferred resistance route, biased towards 215F selection. No HIV clone contained both K65R and T215F/Y mutations, suggesting in vivo antagonism between the two mutations. The once-daily zidovudine usage and high baseline viraemia may also have contributed to rapid selection of HIV with multiple mutations in VFs. PMID- 20008907 TI - Calibration of indium response functions in an Au-In-BSE system up to 800 MeV. AB - Calibration of the response functions of a gold (Au)-indium (In) dual foil Bonner sphere extended (BSE) system was described. The response of the In and Au foil of the system was calculated using MCNPX code with different activation cross sectional libraries: (ACTL and ENDF VI for gold and ACTL and 532DOS2 for In). To verify and correct the calculated response functions the Bonner sphere set (BSS) was irradiated using (252)Cf and (241)AmBe sources of known neutron strengths for neutrons ranging from thermal to 20 MeV, and was irradiated at the 800-MeV neutron beam of the Los Alamos Neutron Science Center. The neutron spectrum of the 800 MeV beam was determined using time-of-flight (TOF) technique. We observed that the uncertainty of activation cross section in the resonance region can result in great uncertainty in the MCNPX-calculated response functions of activation foil-based BSS. The MCNPX-calculated response functions must be corrected using neutron sources of known spectrum and strength. PMID- 20008908 TI - Work-family interference and long-term sickness absence: a longitudinal cohort study. AB - BACKGROUND: Alongside work environment factors, interference between work and domestic life has been proposed as an important explanation for long-term sickness absence, particularly for women. The aim was to investigate the association between work-to-family interference, family-to-work interference and long-term sickness absence among women and men in different family- and work related settings. METHODS: The study population was a random sample of 2867 gainfully employed adults in Sweden aged 25-50. In 2004, telephone interview data were collected that included questions about family, work and health. The outcome measure was having at least one spell of long-term sickness absence (>14 days) in 2005 based on social insurance register data. Associations were analysed by logistic regression. RESULTS: Work-to-family interference was more common than family-to-work interference and more often reported by women. The overall associations with long-term sickness absence were weak. However, after adjustment for age and self-reported health, work-to-family interference was associated with long-term sick leave among men with higher socioeconomic status (odds ratio 2.87; 95% CI 1.36-6.07), and there was also a tendency to association among women bearing the main responsibility for housework and family (1.59; 0.99-2.54). CONCLUSIONS: These findings suggest that work-to-family interference is associated with long-term sickness absence in the working population, but in a gender- and situation-specific manner. Hence, extensive work responsibilities for men, and probably extensive family responsibilities for women, could hamper the balance between work and family and increase the risk of long-term sick leave. Further studies are warranted within this area. PMID- 20008909 TI - Educational health inequalities in former Yugoslavia: evidence from the South East European Social Survey Project. AB - BACKGROUND: An important gap in our knowledge of social inequalities in health is the former Yugoslavia, a region of culturally and historically diverse countries, with recent conflict. The aim of the present paper is to investigate relative and absolute inequalities in self-assessed health in former Yugoslavia (Bosnia Herzegovina, Croatia, Kosovo, Macedonia, Montenegro, Slovenia and Serbia) by sex and education. METHODS: The data source is the South-East European Social Survey Project fielded in December 2003 to Winter 2004, covering the former Yugoslavia with a total sample of 18 481 respondents. Data from Slovenia were obtained from the 2004-wave of the European Social Survey. The health outcome variables were self-reported general health (SRH) and limiting longstanding illness (LLI). RESULTS: Both absolute and relative educational health inequalities were present throughout the former Yugoslavia to a larger or lesser extent, although odds ratios (ORs) for LLI and SRH were not significant for Montenegrin women [LLI OR = 1.12, 95% confidence interval (CI): 0.92-1.37; SRH OR = 1.16, 95% CI: 0.96-1.40] and with respect to the reporting of LLI among Slovenian men (OR = 1.16, 95% CI: 0.96-1.44). Overall, Montenegro held the best position. CONCLUSIONS: The prevalence of poor health and the degree of relative inequality in self-assessed health in the former Yugoslavian countries were similar in order to one another, and to other East European countries during the same period. Influences on subjective health require further elucidation. Further research should study a wider range of health outcomes using larger survey samples and a wider range of cultural and other predictor variables. PMID- 20008910 TI - Expert views on most suitable monetary incentives on food to stimulate healthy eating. AB - BACKGROUND: Pricing strategies are an important component in the marketing mix and may also be useful in stimulating healthier food choices. However, due to competing interests and feasibility problems, the introduction of pricing strategies is complicated. For successfully introducing food pricing strategies, it is essential to explore incentives that are not only promising but also realizable and being approved by different sectors. METHODS: We aimed to assemble a list of pricing strategies by exploring expert views using the Delphi method. Subjects included experts from academia, industry, retail, agriculture, policymakers, consumers and non-governmental organizations. Data were collected in three rounds. In round one, experts designed promising pricing strategies. Based on a time-budget model incorporating Sleep, Leisure, Occupation, Transportation and Home-based activities, these strategies were in the subsequent rounds judged on several criteria. Results were analysed using median and interquartile deviations scores. RESULTS: We found fair consensus levels among experts and a varied list of promising pricing strategies. The panel agreed on the potential success of offering small presents, providing price-cuts on healthy foods and discounting healthier foods more frequently. Also, it was found that experts gave higher rates to pricing strategies for which the implementation responsibilities could be placed elsewhere. CONCLUSION: The resulted list of promising monetary incentives is an essential first step for the future design of pricing strategies. Following this study, it is important to determine how to make solid agreements on responsibility and implementation issues. Also, consumer perceptions regarding the proposed pricing strategies should be studied. PMID- 20008911 TI - The analysis of the ethical, organizational and legal aspects of Polish biobanks activity. AB - BACKGROUND: Biobanks create new opportunities for the public health (Public Health Genomics) but need some regulations. Polish biobanks are in the phase of dynamic transformations aiming at integrating their activity and adjusting them to the international standards. The aim of this article is to present the current standards held in the most important Polish institutions collecting biological samples. METHODS: A survey was conducted by the questionnaire filled by managers and directors of 24 Polish biobanks. Questions concerned obtaining patient informed consent (also a consent to some future researches and access to dates by other organizations), protection of privacy, storage of the samples and the relevant information, the opinion related to the need of establishing Polish national DNA data bank and possibilities of the scientific international cooperation. RESULTS: The written consent to carry out research on the samples is obtained almost everywhere, but 29% of biabanks does not acquire the consent for the future usage of the samples. Almost all inquired banks (93%, n = 22) declare the willingness to start an international cooperation. Most of respondents support the idea of establishing the Polish national DNA bank by the fusion of the already existing banks. CONCLUSION: The inquiry findings point that the current biobanks procedures do not differ to any relevant degree from procedures applied in the West European countries. Many of Polish biobanking organizations are willing to cooperate with foreign sites. It may enhance the European biobanking network and gene-environmental research. PMID- 20008912 TI - Case reports: response to a partial power failure in the operating room. AB - Failure of the power supply to the operating suites is a rare event. When it happens, it is critical that the care team responds in a way that optimizes patient safety. In the event we describe in this case report, only the emergency (red) power outlets failed. The anesthesia care team and other staff in the 8 affected rooms with anesthetized patients generally responded appropriately. The optimal response to these events is not necessarily intuitive. It is important to have advanced planning both within the immediate work area and beyond the operating room all the way into the hospital infrastructure. PMID- 20008914 TI - An anatomical basis for blocking of the deep cervical plexus and cervical sympathetic tract using an ultrasound-guided technique. AB - BACKGROUND: A selective blocking method for the cervical plexus and the cervical sympathetic trunk has not yet been established. METHODS: We performed a detailed examination of the neck anatomy using 28 cadavers. The pattern of local anesthetic distribution after injection in 2 healthy volunteers was imaged using computed tomographic scan. RESULTS: The deep cervical plexus was located in the groove between the longus capitis and scalenus medius muscles. The cervical sympathetic trunk was located on the anteromedial surface of the longus capitis. Although anesthetic injected into the longus capitis was confined to the muscle, it infiltrated into neighboring structures including the C2 to C5 roots and sympathetic trunk. CONCLUSIONS: The longus capitis muscle is a suitable landmark for blocking the cervical plexus and trunk. PMID- 20008913 TI - Modification of the hepatic hemodynamic response to acute changes in PaCO2 by nitric oxide synthase inhibition in rabbits. AB - BACKGROUND: Hypercapnia has been reported to modify liver circulation. The vascular regulations implicated in this response remain partly unknown. METHODS: Using anesthetized and ventilated rabbits, we designed this study to evaluate the hepatic artery and portal vein blood flow velocity adjustments (20 MHz pulsed Doppler) after changes in PaCO2 (by varying the inspiratory fraction of CO2 and to assess the proper role of pH, independent of PaCO2 changes, the role of portal vein CO2, and the effect of nitric oxide synthase inhibition on CO2-induced modifications of hepatic hemodynamics. RESULTS: Increasing PaCO2 from 30.9 +/- 5 mm Hg to 77 +/- 11 mm Hg increased arterial blood pressure by 20% (P < 0.01) and hepatic artery blood flow velocity by 90% (P < 0.05) and decreased aortic blood flow velocity by 15% and portal vein blood flow velocity by 40% (both P < 0.05). Changes in pH (1 mL of 0.1 N hydrochloric acid infusion) or isolated changes in portal vein CO2 at constant PaCO2 induced by CO2 insufflation in an open abdomen had no effect on hepatic hemodynamics. Pretreatment with a nitric oxide synthase inhibitor, N(omega)-nitro-L-arginine (2.5 mg/kg), blunted the systemic response to hypercapnia, whereas the portal modifications persisted, with a largely attenuated hepatic artery blood flow increase. CONCLUSIONS: CO2 per se acts on hepatic blood flow by its systemic effect, probably via chemoreflexes. Nitric oxide does not mediate hepatosplanchnic hemodynamic modifications to acute changes in PaCO2 but may play a permissive role by regulating the amplitude of hepatic vascular response. PMID- 20008915 TI - Perioperative analgesia for forequarter amputation in a child: a dual paravertebral approach. AB - We describe the management of postoperative pain for a 10-year-old girl who underwent forequarter amputation for osteosarcoma of the left humerus. Because the brachial plexus itself was divided and resected during surgery, and the main body part innervated by the nerves from this plexus (the entire upper limb including the scapula and clavicle) was removed, providing analgesia via a brachial plexus block alone would probably not have provided adequate coverage. Because the tissue not resected with this surgery was innervated via the cervical and brachial plexuses and some upper thoracic nerve roots, we elected to combine a perioperative high continuous cervical paravertebral block at the C5 level with a continuous thoracic paravertebral block at the T2 level for postoperative analgesia. Our patient experienced excellent postoperative analgesia and required no narcotics during the immediate postoperative period. PMID- 20008916 TI - Muscle quality, architecture, and activation in cachectic patients with rheumatoid arthritis. AB - OBJECTIVE: To explore muscle-specific force (force per physiological cross sectional area, or PCSA) and muscle activation in cachectic patients with rheumatoid arthritis (RA). METHODS: In 14 muscle-wasted patients with RA and age and sex matched healthy controls, vastus lateralis (VL) force and voluntary activation capacity were assessed during maximal isometric contractions with electromyography and superimposed electrical stimulations. VL PCSA was determined from ultrasound measures of fiber fascicle length (Lf), pennation angle, and volume, together with assessments of body composition by dual energy x-ray absorptiometry and objective physical function. RESULTS: Although patients with RA had reduced physical function, lower muscle mass, and VL volume relative to controls, there were no differences in muscle-specific force and activation. PCSA, force, and pennation angle tended to be lower in RA, with no differences in Lf. CONCLUSION: Muscle-specific force and activation are not compromised and thus are unlikely to contribute to reduced function in cachectic patients with RA. PMID- 20008917 TI - Can tumor necrosis factor inhibitors be safely used in pregnancy? AB - OBJECTIVE: We review available safety data for use of currently approved tumor necrosis factor (TNF) inhibitors during pregnancy and lactation and suggest guidelines for use of these agents among women of reproductive age. Method. Although regulatory agencies encourage the inclusion of pregnant women and those of child-bearing age in randomized controlled trials, pregnant and lactating women have universally been excluded from studies because of unknown or potential risks to the fetus. Thus, strong evidence-based treatment recommendations during pregnancy are usually lacking and safety information is derived from voluntary reports of adverse events during postmarketing surveillance or via uncontrolled, observational studies, reviewed here. RESULTS: Uncommon adverse pregnancy outcomes observed with TNF inhibitor therapy appear to approximate those seen in women not receiving such therapy and may include premature birth, miscarriage, low birthweight, hypertension, and preeclampsia. There are rare reports of fetal malformations or congenital anomalies in patients exposed to TNF inhibitors during conception or pregnancy. However, the incidence of these events appears to be far below the 3% rate of congenital anomalies in the general population. CONCLUSION: If the activity or disease severity precludes the cessation of a TNF inhibitor and/or DMARD, uncontrolled observations suggest that conception and early pregnancy are not adversely affected by use of TNF inhibitors. Nearly 70% of pregnant patients can discontinue their TNF inhibitor early in the pregnancy (or with determination of pregnancy) without augmenting maternal or fetal risks. PMID- 20008918 TI - HIV-associated intracranial aneurysmal vasculopathy in adults. AB - Diffuse fusiform intracranial aneurysms have been reported in children with human immunodeficiency virus (HIV) for over 2 decades, but have only recently been reported in adults with HIV. Although these aneurysms have important clinical implications, their etiology and optimal therapy are unknown. We present a systematic review of diffuse intracranial fusiform aneurysmal vasculopathy in patients who are HIV-positive. We conducted a comprehensive literature search for relevant case reports and reviews published before February 2009. Patients were included if they had HIV infection and radiographic imaging consistent with fusiform aneurysmal vasculopathy. We identify 11 published adult cases of intracranial fusiform aneurysmal vasculopathy and describe 1 unpublished case from our own institution. Available data regarding clinical presentation, characteristic imaging findings, and treatment of this complex syndrome are reviewed. Adults with HIV-associated intracranial aneurysmal vasculopathy typically are significantly immunosuppressed and present with gross neurologic dysfunction. Characteristic radiographic findings include diffuse cerebral fusiform aneurysms with hemorrhage or infarct. Treatment of any active infection followed by the initiation of antiretroviral therapy and corticosteroids may be a reasonable approach in this complex syndrome. PMID- 20008919 TI - Peripheral blood expression profiles of bone morphogenetic proteins, tumor necrosis factor-superfamily molecules, and transcription factor Runx2 could be used as markers of the form of arthritis, disease activity, and therapeutic responsiveness. AB - OBJECTIVE: To assess whether different forms of arthritis and disease activity could be distinguished by peripheral blood expression profiles of bone-regulatory factors including tumor necrosis factor (TNF)-superfamily [TNF-related apoptosis inducing ligand (TRAIL), the Fas ligand (FasL), and the ligand for herpesvirus entry mediator (LIGHT)] and bone morphogenetic protein (BMP)-family members (BMP 2, BMP-4, BMP-6) as well as osteoblast differentiation gene Runx2. METHODS: Blood cells from healthy controls (n = 25) and patients at different disease stages with rheumatoid arthritis (RA; n = 49), osteoarthritis (OA; n = 17), or spondyloarthritis, including ankylosing spondylitis (AS; n = 27) or psoriatic arthritis (PsA; n = 23), were processed for quantitative polymerase chain reaction. Gene expression was assessed in comparison with control samples, correlated with clinical data of different forms of arthritis, and analyzed for discriminative efficacy between groups by receiver-operation characteristic (ROC) curves. Results were confirmed on diagnostic RA (n = 5) and AS (n = 8) samples. RESULTS: BMP-4, BMP-6, and Runx2 expressions were significantly decreased in patients with RA and OA versus controls. Patients with RA also had decreased FasL and LIGHT expression, while patients with AS had increased Runx2 expression. Negative correlation with disease activity was found for BMP-4, FasL, and Runx2 in RA and for Runx2 in PsA, while positive correlation was found for BMP-4 in PsA. Gene expression was higher in the therapy-resistant form of AS (for BMP-4, LIGHT, and Runx2) and in methotrexate-treated patients in RA (for BMP-2 and LIGHT). ROC curve analysis confirmed discrimination between groups, particularly decreased LIGHT and Runx2 for RA and increased Runx2 for AS. CONCLUSION: Our study identified BMP and Runx2 as possible biomarkers of bone metabolism in several forms of arthritis, while lower FasL and LIGHT were associated with RA. Correlation between gene expression and disease activity may be clinically useful in assessing therapeutic effectiveness and disease monitoring. PMID- 20008920 TI - Unresponsiveness to colchicine therapy in patients with familial Mediterranean fever homozygous for the M694V mutation. AB - OBJECTIVE: More than 50 disease-associated mutations of the Mediterranean fever gene (MEFV) have been identified in familial Mediterranean fever (FMF), some of which were shown to have different clinical, diagnostic, prognostic, and therapeutic implications. The aim of our study was to define the frequency of mutation type, genotype-phenotype correlation, and response to colchicine treatment in patients with FMF. METHODS: This study included 222 pediatric FMF patients. All patients were investigated for 6 MEFV mutations. Then patients were divided into 3 groups according to the presence of M694V mutation on both of the alleles (homozygotes), on only 1 allele (heterozygotes), and on none of the alleles, and compared according to their phenotypic characteristics and response to treatment. M694V/M694V was denoted Group A, M694V/Other Group B, and Other/Other, Group C. RESULTS: Complete colchicine response was significantly lower while the rate of unresponsiveness was significantly higher in Group A compared to Groups B and C (p = 0.031, p < 0.001 and p = 0.005, p = 0.029, respectively). No differences except proteinuria were found between the phenotypic features of 3 groups. Group C had the lowest rate of proteinuria development (p = 0.024). All the amyloidosis patients were in Group A. CONCLUSION: Our results indicate that the M694V/M694V mutation is associated with lower response to colchicine treatment. Therefore, patients homozygous for M694V/M694V may be carrying an increased risk for development of amyloidosis. PMID- 20008921 TI - Predictors of damage and survival in patients with Wegener's granulomatosis: analysis of 50 patients. AB - OBJECTIVE: To evaluate damage features and impact on survival by Vasculitis Damage Index (VDI) in a cohort of Turkish patients with Wegener's granulomatosis (WG). METHODS: We enrolled 50 (25 female) patients with WG according to ACR criteria. Birmingham Vasculitis Activity Score (BVAS) and VDI were used to analyze disease activity and damage. RESULTS: Patients had kidney (82%), upper airway (72%), lung (70%), and nervous system (15%) involvement. Median age at diagnosis was 45 years, time to diagnosis was 3.5 months, and total followup time was 35.5 months. All but one patient was positive for antineutrophil cytoplasmic antibodies (ANCA). Mean final dose and duration of corticosteroid and cyclophosphamide was 15 +/- 14 g, 39 +/- 33 months and 36 +/- 34 g, 21 +/- 2 months, respectively. Mean early (e) BVAS were 20.2 +/- 7.1 (4-38) (median 21). Mean e-BVAS and e-VDI scores at presentation and final (f)-VDI scores at last visit were 20.2 +/- 7.1 (4-38), 3.1 +/- 1.7 (median 3) (0-7) and 4.4 +/- 2.6 (0 12), consecutively. Disease related damage was prominent in kidneys (50%) and upper airways (27%). Amenorrhea (90%), cataract (28%), and diabetes (24%) were the most frequent treatment related damages. Rapidly progressive glomerulonephritis at presentation (42%) progressed to endstage renal failure in 20%. Relapses occurred in 25% with mean BVAS of 6.5 +/- 2.3 (4-11). Survival rate was 77% at 37 months. Deaths occurred early (90% in the first year). f-VDI was high in patients who relapsed (6 +/- 3 vs 3.8 +/- 2.1, p = 0.03). Logistic regression analysis demonstrated that age at time of diagnosis and e-VDI were lower in survivors with OR = 0.9 (p = 0.06, 95% CI: 0.78-1) and OR = 0.5 (p = 0.04, 95%CI: 0.25-0.98), respectively. In this cohort, e-VDI score of 5 or more was related to death with 98% sensitivity and 56% specificity (p = 0.004) (CI: 0.66-0.95). CONCLUSION: Disease related damage outweighed treatment related damage in our cohort of predominantly generalized disease activity. Early damage and older age were found to be predictors of final damage and death. PMID- 20008922 TI - Possible reactivation of potential hepatitis B virus occult infection by tumor necrosis factor-alpha blocker in the treatment of rheumatic diseases. AB - OBJECTIVE: To assess the safety of anti-tumor necrosis factor (TNF-alpha) therapy in patients with rheumatic diseases in terms of the reactivation of potential hepatitis B virus (HBV) occult infection. METHODS: Patients who had taken anti TNF-alpha for the treatment of rheumatic diseases from January 2002 to May 2008 were included in the study. In this patient group, we retrospectively investigated a series of serum aminotransferase levels, HBV serologic status, the type of anti-TNF-alpha therapy, duration of the anti-TNF-alpha treatment, and concurrent use of hepatotoxic drugs. RESULTS: A total of 266 cases were documented using 3 serologic markers for HBV infection: HBV surface antigen (HBsAg), HBV surface antibody (HBsAb), and HBV core IgG Ab (HBcAb). Of these, 8 cases had chronic hepatitis B (HBsAg+), 170 cases were HBcAb-negative, and 88 cases were identified as having potential HBV occult infections represented by HBsAg-negative and HBcAb-positive, irrespective of the status of the HBsAb. The frequency of clinically significant (> 2 times normal value) and persistent increase (> 2 consecutive tests) of aminotransferase levels was significantly higher in the group with a potential HBV occult infection compared to the HBcAb negative group. In the multiple logistic regression analysis controlling for various potential confounding factors such as prophylactic anti-tuberculosis medication, methotrexate, nonsteroidal antiinflammatory drugs, and the type of anti-TNF-alpha therapy, only potential HBV occult infection was a significant risk factor for abnormal liver function test (LFT). CONCLUSION: All rheumatic patients who plan to take anti-TNF-alpha treatment should undergo a test for HBV serology, including HBcAb, and have a close followup with an LFT test during therapy. Further prospective studies for hepatitis B viral load using HBV polymerase chain reaction in patients who are HbcAb positive are needed to identify whether the abnormal LFT comes from the reactivation of occult HBV infection. PMID- 20008923 TI - Pulmonary manifestations in Sjogren's syndrome: correlation analysis between chest computed tomographic findings and clinical subsets with poor prognosis in 80 patients. AB - OBJECTIVE: Sjogren's syndrome (SS) has a varied clinical spectrum and has been associated with various chest computed tomography (CT) findings. We sought to delineate the characteristic CT features in various subsets of SS, especially poor prognosis subsets. METHODS: Retrospectively identified 80 never-smoker SS patients [56 primary SS (1-SS), 24 secondary SS (2-SS)] who underwent chest CT at our institution during a 3-year period from 2004 through 2007 were included in this study. Chest CT findings were qualitatively and semiquantitatively analyzed with comparison between 1-SS and 2-SS, and correlation with anti-SSB/La seropositivity and the presence of clonally derived lymphoproliferative disorder (cLPD), which are known to be pathognomonic and prognostic clinical features of SS patients. RESULTS: All patients were women with median age of 60 years. Anti SSB/La antibodies were found in 17 primary SS patients and 4 2-SS patients. Eleven patients with cLPD were identified and all of them had 1-SS. The most frequent CT finding in both types of patients was interlobular septal thickening. Secondary SS was associated with a significantly greater frequency and extent of honeycombing versus 1-SS. Univariate and multivariate analysis showed a significant association between honeycombing and 2-SS. In patients with 1-SS and in the SS group as a whole, we observed independent and significant associations between cysts and anti-SSB/La seropositivity or cLPD. CONCLUSION: Cysts are significantly associated with anti-SSB/La seropositivity and cLPD. The presence of lung cysts revealed by chest CT might be a prognostic clinical feature, a clue, or a predictor of cLPD in patients with SS. PMID- 20008924 TI - Familial Mediterranean fever in Ashkenazi Jews: the mild end of the clinical spectrum. AB - OBJECTIVE: To characterize familial Mediterranean fever (FMF) in Ashkenazi patients, a Jewish subgroup in which FMF has rarely been described before. METHODS: A retrospective analysis, comparing demographic, clinical, and genetic measures of the cohort of Ashkenazi Jewish patients with FMF (n = 57), followed at the National Center for FMF in Israel, to age and sex matched patients of Iraqi Jewish (n = 62) and North African Jewish (NAJ; n = 61) origin. RESULTS: Age at disease onset and diagnosis was earlier in NAJ than among Ashkenazi and Iraqi patients. Family history of FMF was described by only 30% of Ashkenazi patients as opposed to the majority of Iraqi and NAJ patients (p = 0.001). The frequency of abdominal and febrile attacks was similar among the 3 groups, while chest and joint attacks were far less common in Ashkenazi and Iraqi compared to NAJ patients. A good response to colchicine was noted in a similar proportion of Ashkenazi and Iraqi patients (82-84%) as opposed to only 56% of NAJ patients (p = 0.0001). Proteinuria, renal failure, and amyloidosis were most frequent among the NAJ patients (18, 6.6, and 9.8% compared to 5.3, 0, and 3.5% and 1.6, 0, and 0% in Ashkenazi and Iraqi patients, respectively). CONCLUSION: Ashkenazi patients with FMF stand at the mildest end of the clinical spectrum of FMF. This is notwithstanding the tendency for amyloidosis, the frequency of which is not trivial and which deserves particular awareness. PMID- 20008925 TI - Clinical features and prognosis of late-onset systemic lupus erythematosus: results from the 1000 faces of lupus study. AB - OBJECTIVE: There is controversy whether older-onset systemic lupus erythematosus (SLE) is associated with a different, more benign disease course than in younger onset SLE. Our objective was to characterize the clinical features and prognosis of late-onset SLE in a large, multicenter cohort. METHODS: We studied adult-onset lupus in the 1000 Canadian Faces of Lupus cohort (n = 1528) of whom 10.5% had onset at age > or = 50 years versus a control group with onset at < 50 years. RESULTS: Disease duration was different in early- and late-onset groups (15 yrs in early vs 9.3 yrs in late; p < 0.001). Caucasians were represented more in the later-onset SLE group (55.6% vs 74.5%), while Asians and Blacks were more prevalent in the younger group. Younger-onset SLE subjects fulfilled more American College of Rheumatology criteria for SLE (< 50 yrs: 5.98 +/- 1.68; > or = 50 yrs: 5.24 +/- 1.44; p < 0.0001). Despite an equal prevalence of anti-dsDNA, the younger-onset group more often had positive anti-Smith autoantibody, ribonucleoprotein, and hypocomplementemia, and more nephritis, rash, and cytopenias than the older-onset group. However, disease activity and damage accrual were higher in the older-onset group. The older patients received less prednisone and immunosuppressives (current and ever-use). As expected, comorbidity was higher in the older-onset SLE group. CONCLUSION: This study suggests that older age-onset SLE is not benign. There may be an interaction between lupus and age in which, although there is less lupus nephritis in the elderly, more disease activity and damage are present. PMID- 20008926 TI - Trp64Arg polymorphism of the ADRB3 gene predicts hyperuricemia risk in a population from southern Spain. AB - OBJECTIVE: To study the role of Trp64Arg polymorphism of the ADRB3 gene in the risk of developing hyperuricemia in 1051 subjects from southern Spain, with a followup of 6 years. The inclusion of plasma levels of uric acid as a diagnostic criterion to define the metabolic syndrome is under discussion. Genes responsible for insulin resistance could contribute to the development of hyperuricemia. Previous cross-sectional studies have suggested ADRB3 as a possible candidate gene in the development of hyperuricemia and insulin resistance. METHODS: A prospective, population-based, cohort study of 1051 persons examined in 1997-98 and reassessed at a second examination 6 years later. The metabolic phenotype was assessed at baseline and again at the followup. Insulin resistance was measured by homeostasis model assessment. The Trp64Arg polymorphism of ADRB3 was detected by real-time polymerase chain reaction. Subjects were considered normouricemic if their serum uric acid levels were 500 kb in some cases) that contain repressive chromatin marks (e.g., histone H3 Lys 27 trimethylation). The boundaries of macroH2A1-containing domains tend to occur in promoter-proximal regions. Not all promoters, however, serve as macroH2A1 boundaries; many macroH2A1-containing chromatin domains invade the transcribed regions of genes whose products play key roles in development and cell-cell signaling. Surprisingly, the expression of a subset of these genes is positively regulated by macroH2A1. MacroH2A1 also plays a role in augmenting signal-regulated transcription, specifically for genes responsive to serum starvation. Collectively, our results document an unexpected role for macroH2A1 in the escape from heterochromatin-associated silencing and the enhancement of autosomal gene transcription. PMID- 20008928 TI - Alpha3, a transposable element that promotes host sexual reproduction. AB - Theoretical models predict that selfish DNA elements require host sex to persist in a population. Therefore, a transposon that induces sex would strongly favor its own spread. We demonstrate that a protein homologous to transposases, called alpha3, was essential for mating type switch in Kluyveromyces lactis. Mutational analysis showed that amino acids conserved among transposases were essential for its function. During switching, sequences in the 5' and 3' flanking regions of the alpha3 gene were joined, forming a DNA circle, showing that alpha3 mobilized from the genome. The sequences encompassing the alpha3 gene circle junctions in the mating type alpha (MATalpha) locus were essential for switching from MATalpha to MATa, suggesting that alpha3 mobilization was a coupled event. Switching also required a DNA-binding protein, Mating type switch 1 (Mts1), whose binding sites in MATalpha were important. Expression of Mts1 was repressed in MATa/MATalpha diploids and by nutrients, limiting switching to haploids in low-nutrient conditions. A hairpin-capped DNA double-strand break (DSB) was observed in the MATa locus in mre11 mutant strains, indicating that mating type switch was induced by MAT-specific DSBs. This study provides empirical evidence for selfish DNA promoting host sexual reproduction by mediating mating type switch. PMID- 20008929 TI - Transcriptional competence in pluripotency. AB - Embryonic stem (ES) cells possess a globally open, decondensed chromatin structure that, together with trans-acting factors, supports transcriptional competence of developmentally regulated genes. However, our understanding of the mechanisms that establish transcriptional competence of specific genes is limited. In this issue of Genes & Development, Xu and colleagues (pp. 2824-2838) show that tissue-specific enhancers are actively marked by an unmethylated window in ES cells and induced pluripotent stem (iPS) cells. They propose a model and present supporting evidence to demonstrate the active involvement of pioneer transcription factors in this process. This work marks an important step toward the understanding of the mechanisms that define and maintain pluripotency, and calls for the identification of the factors that participate in the establishment of transcriptional competence in pluripotent cells. PMID- 20008930 TI - Overcoming inhibition in the spindle checkpoint. AB - Spindle checkpoint silencing is a critical step during mitosis that initiates chromosome segregation, yet surprisingly little is known about its mechanism. Protein phosphatase I (PP1) was shown recently to be a key player in this process, and in this issue of Genes & Deverlopment, Akiyoshi and colleagues (pp. 2887-2899) identify budding yeast Fin1p as a kinetochore-localized regulator of PP1 activity toward checkpoint targets. Here we review recent mechanistic insights and propose a working model for spindle checkpoint silencing. PMID- 20008931 TI - Genetic dissection of the miR-17~92 cluster of microRNAs in Myc-induced B-cell lymphomas. AB - The miR-17 approximately 92 cluster is frequently amplified or overexpressed in human cancers and has emerged as the prototypical oncogenic polycistron microRNA (miRNA). miR-17 approximately 92 is a direct transcriptional target of c-Myc, and experiments in a mouse model of B-cell lymphomas have shown cooperation between these two oncogenes. However, both the molecular mechanism underlying this cooperation and the individual miRNAs that are responsible for it are unknown. By using a conditional knockout allele of miR-17 approximately 92, we show here that sustained expression of endogenous miR-17 approximately 92 is required to suppress apoptosis in Myc-driven B-cell lymphomas. Furthermore, we show that among the six miRNAs that are encoded by miR-17 approximately 92, miR-19a and miR 19b are absolutely required and largely sufficient to recapitulate the oncogenic properties of the entire cluster. Finally, by combining computational target prediction, gene expression profiling, and an in vitro screening strategy, we identify a subset of miR-19 targets that mediate its prosurvival activity. PMID- 20008932 TI - Neuronal SIRT1 regulates endocrine and behavioral responses to calorie restriction. AB - Mammalian life span can be extended by both calorie restriction (CR) and mutations that diminish somatotropic signaling. Sirt1 is a mediator of many effects of CR in mammals, but any role in controlling somatotropic signaling has not been shown. Since the somatotropic axis is controlled by the brain, we created mice lacking Sirt1 specifically in the brain and examined the impacts of this manipulation on somatotropic signaling and the CR response. These mutant mice displayed defects in somatotropic signaling when fed ad libitum, and defects in the endocrine and behavioral responses to CR. We conclude that Sirt1 in the brain is a link between somatotropic signaling and CR in mammals. PMID- 20008933 TI - A novel histone fold domain-containing protein that replaces TAF6 in Drosophila SAGA is required for SAGA-dependent gene expression. AB - The histone acetyltransferase complex SAGA is well characterized as a coactivator complex in yeast. In this study of Drosophila SAGA (dSAGA), we describe three novel components that include an ortholog of Spt20, a potential ortholog of Sgf73/ATXN7, and a novel histone fold protein, SAF6 (SAGA factor-like TAF6). SAF6, which binds directly to TAF9, functions analogously in dSAGA to TAF6/TAF6L in the yeast and human SAGA complexes, respectively. Moreover, TAF6 in flies is restricted to TFIID. Mutations in saf6 disrupt SAGA-regulated gene expression without disrupting acetylated or ubiquitinated histone levels. Thus, SAF6 is essential for SAGA coactivator function independent of the enzymatic activities of the complex. PMID- 20008934 TI - Transcriptional competence and the active marking of tissue-specific enhancers by defined transcription factors in embryonic and induced pluripotent stem cells. AB - We reported previously that well-characterized enhancers but not promoters for typical tissue-specific genes, including the classic Alb1 gene, contain unmethylated CpG dinucleotides and evidence of pioneer factor interactions in embryonic stem (ES) cells. These properties, which are distinct from the bivalent histone modification domains that characterize the promoters of genes involved in developmental decisions, raise the possibility that genes expressed only in differentiated cells may need to be marked at the pluripotent stage. Here, we demonstrate that the forkhead family member FoxD3 is essential for the unmethylated mark observed at the Alb1 enhancer in ES cells, with FoxA1 replacing FoxD3 following differentiation into endoderm. Up-regulation of FoxD3 and loss of CpG methylation at the Alb1 enhancer accompanied the reprogramming of mouse embryonic fibroblasts (MEFs) into induced pluripotent stem (iPS) cells. Studies of two genes expressed in specific hematopoietic lineages revealed that the establishment of enhancer marks in ES cells and iPS cells can be regulated both positively and negatively. Furthermore, the absence of a pre-established mark consistently resulted in resistance to transcriptional activation in the repressive chromatin environment that characterizes differentiated cells. These results support the hypothesis that pluripotency and successful reprogramming may be critically dependent on the marking of enhancers for many or all tissue specific genes. PMID- 20008935 TI - miR-19 is a key oncogenic component of mir-17-92. AB - Recent studies have revealed the importance of multiple microRNAs (miRNAs) in promoting tumorigenesis, among which mir-17-92/Oncomir-1 exhibits potent oncogenic activity. Genomic amplification and elevated expression of mir-17-92 occur in several human B-cell lymphomas, and enforced mir-17-92 expression in mice cooperates with c-myc to promote the formation of B-cell lymphomas. Unlike classic protein-coding oncogenes, mir-17-92 has an unconventional gene structure, where one primary transcript yields six individual miRNAs. Here, we functionally dissected the individual components of mir-17-92 by assaying their tumorigenic potential in vivo. Using the Emu-myc model of mouse B-cell lymphoma, we identified miR-19 as the key oncogenic component of mir-17-92, both necessary and sufficient for promoting c-myc-induced lymphomagenesis by repressing apoptosis. The oncogenic activity of miR-19 is at least in part due to its repression of the tumor suppressor Pten. Consistently, miR-19 activates the Akt-mTOR (mammalian target of rapamycin) pathway, thereby functionally antagonizing Pten to promote cell survival. Our findings reveal the essential role of miR-19 in mediating the oncogenic activity of mir-17-92, and implicate the functional diversity of mir-17 92 components as the molecular basis for its pleiotropic effects during tumorigenesis. PMID- 20008936 TI - Fusion of nearby inverted repeats by a replication-based mechanism leads to formation of dicentric and acentric chromosomes that cause genome instability in budding yeast. AB - Large-scale changes (gross chromosomal rearrangements [GCRs]) are common in genomes, and are often associated with pathological disorders. We report here that a specific pair of nearby inverted repeats in budding yeast fuse to form a dicentric chromosome intermediate, which then rearranges to form a translocation and other GCRs. We next show that fusion of nearby inverted repeats is general; we found that many nearby inverted repeats that are present in the yeast genome also fuse, as does a pair of synthetically constructed inverted repeats. Fusion occurs between inverted repeats that are separated by several kilobases of DNA and share >20 base pairs of homology. Finally, we show that fusion of inverted repeats, surprisingly, does not require genes involved in double-strand break (DSB) repair or genes involved in other repeat recombination events. We therefore propose that fusion may occur by a DSB-independent, DNA replication-based mechanism (which we term "faulty template switching"). Fusion of nearby inverted repeats to form dicentrics may be a major cause of instability in yeast and in other organisms. PMID- 20008937 TI - Nearby inverted repeats fuse to generate acentric and dicentric palindromic chromosomes by a replication template exchange mechanism. AB - Gene amplification plays important roles in the progression of cancer and contributes to acquired drug resistance during treatment. Amplification can initiate via dicentric palindromic chromosome production and subsequent breakage fusion-bridge cycles. Here we show that, in fission yeast, acentric and dicentric palindromic chromosomes form by homologous recombination protein-dependent fusion of nearby inverted repeats, and that these fusions occur frequently when replication forks arrest within the inverted repeats. Genetic and molecular analyses suggest that these acentric and dicentric palindromic chromosomes arise not by previously described mechanisms, but by a replication template exchange mechanism that does not involve a DNA double-strand break. We thus propose an alternative mechanism for the generation of palindromic chromosomes dependent on replication fork arrest at closely spaced inverted repeats. PMID- 20008938 TI - Stn1-Ten1 is an Rpa2-Rpa3-like complex at telomeres. AB - In budding yeast, Cdc13, Stn1, and Ten1 form a heterotrimeric complex (CST) that is essential for telomere protection and maintenance. Previous bioinformatics analysis revealed a putative oligonucleotide/oligosaccharide-binding (OB) fold at the N terminus of Stn1 (Stn1N) that shows limited sequence similarity to the OB fold of Rpa2, a subunit of the eukaryotic ssDNA-binding protein complex replication protein A (RPA). Here we present functional and structural analyses of Stn1 and Ten1 from multiple budding and fission yeast. The crystal structure of the Candida tropicalis Stn1N complexed with Ten1 demonstrates an Rpa2N-Rpa3 like complex. In both structures, the OB folds of the two components pack against each other through interactions between two C-terminal helices. The structure of the C-terminal domain of Saccharomyces cerevisiae Stn1 (Stn1C) was found to comprise two related winged helix-turn-helix (WH) motifs, one of which is most similar to the WH motif at the C terminus of Rpa2, again supporting the notion that Stn1 resembles Rpa2. The crystal structure of the fission yeast Schizosaccharomyces pombe Stn1N-Ten1 complex exhibits a virtually identical architecture as the C. tropicalis Stn1N-Ten1. Functional analyses of the Candida albicans Stn1 and Ten1 proteins revealed critical roles for these proteins in suppressing aberrant telomerase and recombination activities at telomeres. Mutations that disrupt the Stn1-Ten1 interaction induce telomere uncapping and abolish the telomere localization of Ten1. Collectively, our structural and functional studies illustrate that, instead of being confined to budding yeast telomeres, the CST complex may represent an evolutionarily conserved RPA-like telomeric complex at the 3' overhangs that works in parallel with or instead of the well-characterized POT1-TPP1/TEBPalpha-beta complex. PMID- 20008939 TI - Human POT1 is required for efficient telomere C-rich strand replication in the absence of WRN. AB - Mechanisms of telomere replication remain poorly defined. It has been suggested that G-rich telomeric strand replication by lagging mechanisms requires, in a stochastic way, the WRN protein. Here we show that this requirement is more systematic than previously thought. Our data are compatible with a situation in which, in the absence of WRN, DNA synthesis at replication forks is uncoupled, thus allowing replication to continue on the C strand, while single G strands accumulate. We also show that in cells in which both WRN and POT1 are limiting, both G- and C-rich telomeric strands shorten, suggesting a complete replication block. Under this particular condition, expression of a fragment spanning the two POT1-OB (oligonucleotide-binding) fold domains is able to restore C (but not G) strand replication, suggesting that binding of POT1 to the lagging strand allows DNA synthesis uncoupling in the absence of WRN. Furthermore, in vitro experiments indicate that purified POT1 has a higher affinity for the telomeric G-rich strand than purified RPA. We propose a model in which the relative enrichments of POT1 versus RPA on the telomeric lagging strand allows or does not allow uncoupling of DNA synthesis at the replication fork. Our study reveals an unanticipated role for hPOT1 during telomere replication. PMID- 20008940 TI - Boron nutrition of cultured tobacco BY-2 cells. VI. Calcium is involved in early responses to boron deprivation. AB - Boron (B) deprivation induces various responses in plant cells, some of which can be observed very early. However, it has been unknown what kind of signal is generated by the stress. We found that B deprivation induced the expression of stress-responsive genes within 1 h in suspension-cultured tobacco BY-2 cells. The induction was largely suppressed by withholding medium Ca(2+) or by adding a Ca(2+) channel blocker. Analysis using aequorin-expressing cells showed that B deprived cells took up more Ca(2+) than control cells. These results suggest that Ca(2+) influx plays a role in B deprivation stress signaling. PMID- 20008941 TI - A randomized, controlled trial of computer-assisted cognitive remediation for schizophrenia. AB - OBJECTIVE: There is considerable interest in cognitive remediation for schizophrenia, but its essential components are still unclear. The goal of the current study was to develop a broadly targeted computer-assisted cognitive remediation program and conduct a rigorous clinical trial in a large group of schizophrenia patients. METHOD: Sixty-nine people with schizophrenia or schizoaffective disorder were randomly assigned to 36 sessions of computer assisted cognitive remediation or an active control condition. Remediation broadly targeted cognitive and everyday performance by providing supportive, graduated training and practice in selecting, executing, and monitoring cognitive operations. It used engaging computer-based cognitive exercises and one-on-one training. A total of 61 individuals (34 in remediation group, 27 in control group) engaged in treatment, completed posttreatment assessments, and were included in intent-to-treat analyses. Primary outcomes were remediation exercise metrics, neuropsychological composites (episodic memory, working memory, attention, executive functioning, and processing speed), and proxy measures of community functioning. RESULTS: Regression modeling indicated that performance on eight of 10 exercise metrics improved significantly more in the remediation condition than in the control condition. The mean effect size, favoring the remediation condition, was 0.53 across all 10 metrics. However, there were no significant benefits of cognitive remediation on any neuropsychological or functional outcome measure, either immediately after treatment or at the 3-month follow-up. CONCLUSIONS: Cognitive remediation for people with schizophrenia was effective in improving performance on computer exercises, but the benefits of training did not generalize to broader neuropsychological or functional outcome measures. The evidence for this treatment approach remains mixed. PMID- 20008942 TI - Suicide in HIV-infected individuals and the general population in Switzerland, 1988-2008. AB - OBJECTIVE: High rates of suicide have been described in HIV-infected patients, but it is unclear to what extent the introduction of highly active antiretroviral therapy (HAART) has affected suicide rates. The authors examined time trends and predictors of suicide in the pre-HAART (1988-1995) and HAART (1996-2008) eras in HIV-infected patients and the general population in Switzerland. METHOD: The authors analyzed data from the Swiss HIV Cohort Study and the Swiss National Cohort, a longitudinal study of mortality in the Swiss general population. The authors calculated standardized mortality ratios comparing HIV-infected patients with the general population and used Poisson regression to identify risk factors for suicide. RESULTS: From 1988 to 2008, 15,275 patients were followed in the Swiss HIV Cohort Study for a median duration of 4.7 years. Of these, 150 died by suicide (rate 158.4 per 100,000 person-years). In men, standardized mortality ratios declined from 13.7 (95% CI=11.0-17.0) in the pre-HAART era to 3.5 (95% CI=2.5-4.8) in the late HAART era. In women, ratios declined from 11.6 (95% CI=6.4-20.9) to 5.7 (95% CI=3.2-10.3). In both periods, suicide rates tended to be higher in older patients, in men, in injection drug users, and in patients with advanced clinical stage of HIV illness. An increase in CD4 cell counts was associated with a reduced risk of suicide. CONCLUSIONS: Suicide rates decreased significantly with the introduction of HAART, but they remain above the rate observed in the general population, and risk factors for suicide remain similar. HIV-infected patients remain an important target group for suicide prevention. PMID- 20008944 TI - The development of the Feighner criteria: a historical perspective. AB - This essay outlines the historical context in which the Feighner criteria emerged; reconstructs, as far as possible, the process by which the criteria were developed; and traces the influence the criteria had on subsequent developments in American psychiatry. In the 1950s, when American psychiatry under psychoanalytic dominance had little interest in psychiatric diagnosis, Edwin Gildea recruited to the Department of Psychiatry at Washington University faculty who advocated a medical model for psychiatry in which diagnosis had a central role. In 1967, at the urging of the then-resident John Feighner, a discussion group led by Eli Robins and including Sam Guze, George Winokur, Robert Woodruff, and Rod Munoz began meeting with the initial goal of writing a review of prior key contributions to psychiatric diagnosis. In their meetings over the next year, the task soon shifted to the development of a set of new diagnostic criteria. For three diagnoses, major depression, antisocial personality disorder, and alcoholism, the authors could identify the original criteria from which this group worked and the rationale for many of the changes they introduced. Published in 1972, the Feighner criteria were soon widely cited and used in research, and they formed the basis for the development of the Research Diagnostic Criteria, which in turn were central to the development of DSM-III. The team that developed the Feighner criteria made three key contributions to psychiatry: the systematic use of operationalized diagnostic criteria; the reintroduction of an emphasis on illness course and outcome; and an emphasis on the need, whenever possible, to base diagnostic criteria on empirical evidence. PMID- 20008943 TI - Association of FKBP5 polymorphisms with suicidal events in the Treatment of Resistant Depression in Adolescents (TORDIA) study. AB - OBJECTIVE: The authors sought to assess the relationship between candidate genes and two clinical outcomes, namely, symptomatic improvement and the occurrence of suicidal events, in a sample of treatment-resistant depressed adolescents. METHOD: A subsample of depressed adolescents participating in the Treatment of SSRI-Resistant Depression in Adolescents (TORDIA) trial, 155 of whom were of European origin, were genotyped with respect to 21 polymorphisms on 12 genes that have a reported association with depression, treatment response, or suicidal events. Participants had not responded to a previous adequate trial with an antidepressant and were randomized to receive either another selective serotonin reuptake inhibitor or venlafaxine, with or without cognitive-behavioral therapy (CBT). Single-nucleotide polymorphism (SNP) analyses were conducted using PLINK with permutation procedures. RESULTS: No relationship was observed between any polymorphism and response to treatment. The FKBP5 (which codes for a protein causing subsensitivity of the glucocorticoid receptor) rs1360780TT and rs3800373GG genotypes were associated with suicidal events (N=18), even after controlling for treatment effects and relevant covariates. These two SNPs were in significant linkage disequilibrium (r=0.91). CONCLUSIONS: The FKBP5 genotypes associated with suicidal events in this study have been reported by others to cause the greatest degree of glucocorticoid receptor subsensitivity. These results are consistent with those of other studies linking alterations in the hypothalamic-pituitary-adrenal axis with suicidal behavior. The small number of events and lack of a placebo condition make these results preliminary. Replication with a larger sample and a placebo condition is needed to assess whether these events are related to treatment. PMID- 20008945 TI - A randomized trial of medical care management for community mental health settings: the Primary Care Access, Referral, and Evaluation (PCARE) study. AB - OBJECTIVE: Poor quality of healthcare contributes to impaired health and excess mortality in individuals with severe mental disorders. The authors tested a population-based medical care management intervention designed to improve primary medical care in community mental health settings. METHOD: A total of 407 subjects with severe mental illness at an urban community mental health center were randomly assigned to either the medical care management intervention or usual care. For individuals in the intervention group, care managers provided communication and advocacy with medical providers, health education, and support in overcoming system-level fragmentation and barriers to primary medical care. RESULTS: At a 12-month follow-up evaluation, the intervention group received an average of 58.7% of recommended preventive services compared with a rate of 21.8% in the usual care group. They also received a significantly higher proportion of evidence-based services for cardiometabolic conditions (34.9% versus 27.7%) and were more likely to have a primary care provider (71.2% versus 51.9%). The intervention group showed significant improvement on the SF-36 mental component summary (8.0% [versus a 1.1% decline in the usual care group]) and a nonsignificant improvement on the SF-36 physical component summary. Among subjects with available laboratory data, scores on the Framingham Cardiovascular Risk Index were significantly better in the intervention group (6.9%) than the usual care group (9.8%). CONCLUSIONS: Medical care management was associated with significant improvements in the quality and outcomes of primary care. These findings suggest that care management is a promising approach for improving medical care for patients treated in community mental health settings. PMID- 20008946 TI - Combination of antidepressant medications from treatment initiation for major depressive disorder: a double-blind randomized study. AB - OBJECTIVE: Various classes of antidepressant medications generally induce remission of major depressive disorder in only about one-third of patients. In a previous study using mirtazapine or paroxetine alone or in combination from treatment initiation, the rate of patients who remitted within a 6-week period was twice that of patients using either drug alone. In this double-blind study, the authors sought to produce evidence for the superiority of different combinations of antidepressant drugs from treatment initiation. METHOD: Patients (N=105) meeting DSM-IV criteria for major depressive disorder were randomly assigned to receive, from treatment initiation, either fluoxetine monotherapy (20 mg/day) or mirtazapine (30 mg/day) in combination with fluoxetine (20 mg/day), venlafaxine (225 mg/day titrated in 14 days), or bupropion (150 mg/day) for 6 weeks. The primary outcome measure was the Hamilton Depression Rating Scale (HAM D) score. RESULTS: The overall dropout rate was 15%, without notable differences among the four groups. Compared with fluoxetine monotherapy, all three combination groups had significantly greater improvements on the HAM-D. Remission rates (defined as a HAM-D score of 7 or less) were 25% for fluoxetine, 52% for mirtazapine plus fluoxetine, 58% for mirtazapine plus venlafaxine, and 46% for mirtazapine plus bupropion. Among patients who had a marked response, double blind discontinuation of one agent produced a relapse in about 40% of cases. CONCLUSIONS: The combination treatments were as well tolerated as fluoxetine monotherapy and more clinically effective. The study results, which add to a growing body of evidence, suggest that use of antidepressant combinations from treatment initiation may double the likelihood of remission compared with use of a single medication. PMID- 20008947 TI - Olanzapine long-acting injection: a 24-week, randomized, double-blind trial of maintenance treatment in patients with schizophrenia. AB - OBJECTIVE: The purpose of the present study was to evaluate the efficacy and tolerability of olanzapine long-acting injection for maintenance treatment of schizophrenia. METHOD: Outpatients with schizophrenia who had maintained stability on an oral regimen of olanzapine (10, 15, or 20 mg/day) for 4 to 8 weeks were randomly assigned to 24 weeks of double-blind treatment with "low" (150 mg every 2 weeks; N=140), "medium" (405 mg every 4 weeks; N=318), or "high" (300 mg every 2 weeks; N=141) doses of olanzapine long-acting injection; a very low reference dose of olanzapine long-acting injection (45 mg every 4 weeks; N=144); or their stabilized dose of oral olanzapine (N=322). Rates of and time to psychotic exacerbation were estimated using Kaplan-Meier methodology. RESULTS: At 24 weeks, the majority of oral olanzapine-treated patients (93%), as well as most olanzapine long-acting injection-treated patients receiving high (95%), medium (90%), low (84%), and very low doses (69%), remained exacerbation free, with the therapeutic 4-week regimen (medium dose) and pooled 2-week regimen (low and high doses) demonstrating efficacy similar to that of oral olanzapine as well as to each other. The three standard long-acting doses were superior to the very low reference dose based on time to exacerbation. Incidence of weight gain > or = 7% of baseline was 21% for oral olanza-pine compared with 21%, 15%, 16%, and 8% for the high, medium, low, and very low olanzapine long-acting treatment groups, respectively. No clinically significant differences were observed between the long-acting injection and oral olanzapine groups in general safety parameters. Few injection-site reactions occurred (3%). Two patients experienced sedation and delirium consistent with olanzapine overdose following possible accidental intravascular injection. CONCLUSIONS: Olanzapine long-acting injection was efficacious in maintenance treatment of schizophrenia for up to 24 weeks, with a safety profile similar to oral olanzapine except for injection-related adverse events. PMID- 20008953 TI - Plant adaptations and microbial processes in wetlands. PMID- 20008948 TI - Impact of substance use disorders on recovery from episodes of depression in bipolar disorder patients: prospective data from the Systematic Treatment Enhancement Program for Bipolar Disorder (STEP-BD). AB - OBJECTIVE: Bipolar disorder is highly comorbid with substance use disorders, and this comorbidity may be associated with a more severe course of illness, but the impact of comorbid substance abuse on recovery from major depressive episodes in these patients has not been adequately examined. The authors hypothesized that comorbid drug and alcohol use disorders would be associated with longer time to recovery in patients with bipolar disorder. METHOD: Subjects (N=3,750) with bipolar I or bipolar II disorder enrolled in the Systematic Treatment Enhancement Program for Bipolar Disorder (STEP-BD) were followed prospectively for up to 2 years. Prospectively observed depressive episodes were identified for this analysis. Subjects with a past or current drug or alcohol use disorder were compared with those with no history of drug or alcohol use disorders on time to recovery from depression and time until switch to a manic, hypomanic, or mixed episode. RESULTS: During follow up, 2,154 subjects developed a new-onset major depressive episode; of these, 457 subjects switched to a manic, hypomanic, or mixed episode prior to recovery. Past or current substance use disorder did not predict time to recovery from a depressive episode relative to no substance use comorbidity. However, those with current or past substance use disorder were more likely to experience switch from depression directly to a manic, hypomanic, or mixed state. CONCLUSIONS: Current or past substance use disorders were not associated with longer time to recovery from depression but may contribute to greater risk of switch into manic, mixed, or hypomanic states. The mechanism conferring this increased risk merits further study. PMID- 20008954 TI - Proceedings of the 2009 National Toxicology Program Satellite Symposium. AB - The National Toxicology Program (NTP) Satellite Symposium is a one-day meeting that is held in conjunction with the annual Society of Toxicologic Pathology (STP) meeting. The topic of the 2009 Symposium was "Tumor Pathology and INHAND (International Harmonization of Nomenclature and Diagnostic Criteria for Lesions in Rats and Mice) Nomenclature." The goal of this article is to provide summaries of each speaker's presentation, including the diagnostic or nomenclature issues that were presented, along with a few select images that were used for voting. The results of the voting process and interesting points of discussion that were raised during the presentation are also provided. A supplemental file with voting choices and voting results for each case presented at the symposium is available at http://tpx.sagepub.com/supplemental. PMID- 20008955 TI - Interviews and internet forums: a comparison of two sources of qualitative data. AB - We report a comparative keyword analysis of interviews and Internet postings involving people with breast and prostate cancer and discussion of sexual health. Interviewees produce retrospective accounts, their content guided by interviewers' questions, which might elicit rich biographical and contextual details. Internet exchanges concern participants' current experiences and contain detailed accounts of disease processes, medical procedures, bodily processes, and, in the case of sexual health, sexual practices. They are used by participants to exchange information and support in a relatively anonymous context. Because of the ease with which large amounts of such archived Internet materials can be accessed and analyzed, this source has considerable potential for direct observation of illness experiences, although some disadvantages also exist. This reverses an earlier situation where observational research was more laborious than qualitative interviews. Observational material for research purposes is, through the Internet, now easy to obtain and produces naturalistic data. PMID- 20008957 TI - Parstatin(1-26): the putative signal peptide of protease-activated receptor 1 confers potent protection from myocardial ischemia-reperfusion injury. AB - Parstatin, the N-terminal 41-amino-acid peptide cleaved by thrombin from the protease-activated receptor 1, protects against rat myocardial ischemia and reperfusion injury. In this study, we determined that the parstatin fragment 1 26, the putative signal peptide of protease-activated receptor 1, contains the functional domain of parstatin. We assessed a synthesized parstatin(1-26) peptide in an in vivo rat model of myocardial regional ischemia-reperfusion injury (n = 6/group). Infarct size in control rat hearts was 58 +/- 1% area at risk. Parstatin(1-26) was able to reduce infarct size to 13 +/- 1% (P < 0.001) and 22 +/- 1% area at risk (P < 0.01) when given before or after reperfusion. The infarct-sparing effects of parstatin(1-26) were abolished by inhibition of G(i) proteins (pertussis toxin), phosphoinositide 3-kinase/Akt (wortmannin), nitric oxide synthase (NOS; N(G)-monomethyl-l-arginine), soluble guanylyl cyclase [1H [1,2,4]oxadiazolo[4,3-a]quinoxalin-1-one (ODQ)], and sarcolemmal and mitochondrial K(ATP) channels [glibenclamide, 5-hydroxydecanoic acid, and sodium (5-(2-(5-chloro-2-methoxybenzamido)ethyl)-2-methoxyphenylsulfonyl) (methylcarbamothioyl)amide (HMR 1098)]. Parstatin(1-26) cardioprotection was also abolished by atractyloside, a mitochondrial permeability transition pore (mPTP) opener. The inhibitors and opener alone had no effect on infarct size. Furthermore, preischemic treatment with parstatin(1-26) increased Akt and endothelial NOS phosphorylation at the time of reperfusion. After a 120-min reperfusion, parstatin(1-26) increased nitric oxide levels (12 +/- 0.4 to 17 +/- 0.9 mmol/g tissue) and cyclic GMP levels (87 +/- 21 to 395 +/- 36 pmol/g tissue). Parstatin(1-26) treatment either before or after ischemia results in an extremely efficacious protection against ischemia-reperfusion injury that depends on a G(i) protein-mediated pathway involving mPTP, the end effector of the preconditioning pathway. This suggests that parstatin(1-26) has a potential therapeutic role in the treatment of ischemia and reperfusion injury. PMID- 20008956 TI - Identifying a resistance determinant for the antimitotic natural products disorazole C1 and A1. AB - Disorazoles are macrocyclic polyketides first isolated from the fermentation broth of the myxobacterium Sorangium cellulosum. Both the major fermentation product disorazole A(1) and its much rarer companion disorazole C(1) exhibit potent cytotoxic activity against many human tumor cells. Furthermore, the disorazoles appear to bind tubulin uniquely among known antimitotic agents, promoting apoptosis or premature senescence. It is uncertain what conveys tumor cell sensitivity to these complex natural products. Therefore, we generated and characterized human tumor cells resistant to disorazole C(1). Resistant cells proved exceedingly difficult to generate and required single step mutagenesis with chronic stepwise exposure to increasing concentrations of disorazole C(1). Compared with wild-type HeLa cells, disorazole C(1)-resistant HeLa/DZR cells were 34- and 8-fold resistant to disorazole C(1) and disorazole A(1) growth inhibition, respectively. HeLa/DZR cells were also remarkably cross-resistant to vinblastine (280-fold), paclitaxel (2400-fold), and doxorubicin (47-fold) but not cisplatin, suggesting a multidrug-resistant phenotype. Supporting this hypothesis, MCF7/MDR cells were 10-fold cross-resistant to disorazole C(1). HeLa/DZR disorazole resistance was not durable in the absence of chronic compound exposure. Verapamil reversed HeLa/DZR resistance to disorazole C(1) and disorazole A(1). Moreover, HeLa/DZR cells expressed elevated levels of the drug resistance ATP-binding cassette ABCB1 transporter. Loss of ABCB1 by incubation with short interfering RNA restored sensitivity to the disorazoles. Thus, the multidrug resistance transporter ABCB1 can affect the cytotoxicity of both disorazole C(1) and A(1). Disorazole C(1), however, retained activity against cells resistant against the clinically used microtubule-stabilizing agent epothilone B. PMID- 20008958 TI - Histamine potentiates N-methyl-D-aspartate receptors by interacting with an allosteric site distinct from the polyamine binding site. AB - Histamine potentiates activation of native and recombinant N-methyl-d-aspartate receptors (NMDARs), but its mechanisms of action and physiological functions in the brain remain controversial. Using four different models, we have further investigated the histamine-induced potentiation of various NMDAR-mediated responses. In single cultured hippocampal neurons, histamine potentiated NMDA currents. It also potentiated the NMDA-induced increase in intracellular calcium in the absence, as well as with saturating concentrations, of exogenous d-serine, indicating both glycine-dependent and glycine-independent components of its effect. In rat hippocampal synaptosomes, histamine strongly potentiated NMDA induced [(3)H]noradrenaline release. The profile of this response contained several signatures of the histamine-mediated effect at neuronal or recombinant NMDARs. It was NR2B-selective, being sensitive to micromolar concentrations of ifenprodil. It was reproduced by tele-methylhistamine, the metabolite of histamine in brain, and it was antagonized by impromidine, an antagonist/inverse agonist of histamine on NMDA currents. Up to now, histamine was generally considered to interact with the polyamine site of the NMDAR. However, spermine did not enhance NMDA-induced [(3)H]noradrenaline release from synaptosomes, and the potentiation of the same response by tele-methylhistamine was not antagonized by the polyamine antagonist arcaine. In hippocampal membranes, like spermine, tele-methylhistamine enhanced [(3)H]dl-(E)-2-amino-4-propyl-5-phosphono-3 pentenoic acid (CGP39653) binding to the glutamate site. In contrast, spermine increased nonequilibrium [(3)H]5H-dibenzo[a,d]cyclohepten-5,10-imine (dizocilpine maleate; MK-801) binding, and suppressed [(3)H]ifenprodil binding, whereas histamine and tele-methylhistamine had no effect. In conclusion, the histamine induced potentiation of NMDARs occurs in the brain under normal conditions. Histamine does not bind to the polyamine site, but to a distinct entity, the so called histamine site of the NMDAR. PMID- 20008963 TI - A polysaccharide, MDG-1, induces S1P1 and bFGF expression and augments survival and angiogenesis in the ischemic heart. AB - Ophiopogon japonicus is a traditional Chinese medicine used to treat cardiovascular disease. Recent studies have confirmed its beneficial properties, but not the mechanism of action. Herein, we investigate the anti-ischemic properties of a water-soluble beta-d-fructan (MDG-1) from Ophiopogon japonicus, and assess the cytoprotective and proangiogenic effects of MDG-1. MDG-1 protects cardiomyocyte and microvascular endothelial cells (HMEC-1) against oxygen glucose deprivation (OGD)-induced cell death, as well as protect myocardial cells from ischemia-induced death occurring after coronary artery ligation in rats. Meanwhile, MDG-1 stimulates the differentiation of HMEC-1 cells into capillary like structures in vitro and functions as a chemoattractant in migration assays, and promotes neovascularization in ischemic myocardium. In addition, MDG-1 upregulates sphingosine kinase 1 and sphingosine-1-phosphate (S1P) receptor 1 expression. Both MDG-1 and S1P induce basic fibroblast growth factor (bFGF) expression in HMEC-1 cells. Further study revealed that both MDG-1 and S1P induce Akt and ERK phosphorylation in a dose- and time-dependent manner, an effect that is attenuated by pre-treatment with either the Akt inhibitor wortmannin or the ERK inhibitor PD98059, and MDG-1 can also induce eNOS phosphorylation and increases in production of NO. These data indicate that MDG-1 presented remarkable anti-ischemic activity and protects cardiomyocyte and HMEC-1 cells from ischemia-induced cell damage by inducing S1P1 and bFGF cytoprotective and proangiogenic effects via the S1P/bFGF/Akt/ERK/eNOS signaling pathway. PMID- 20008964 TI - Who am I without you? The influence of romantic breakup on the self-concept. AB - Romantic relationships alter the selves of the individuals within them. Partners develop shared friends and activities and even overlapping self-concepts. This intertwining of selves may leave individuals' self-concepts vulnerable to change if the relationship ends. The current research examines several different types of self-concept change that could occur after a breakup and their relation to emotional distress. Across three studies, using varied methodologies, the authors examined change in both the content (Study 1a and 1b) and the structure of the self-concept, specifically, reduced self-concept clarity (Studies 1 through 3). As predicted, individuals experienced self-concept content change and reduced self-concept clarity post-breakup. Additionally, reduced clarity uniquely predicted post-breakup emotional distress. PMID- 20008965 TI - System justification, the denial of global warming, and the possibility of "system-sanctioned change". AB - Despite extensive evidence of climate change and environmental destruction, polls continue to reveal widespread denial and resistance to helping the environment. It is posited here that these responses are linked to the motivational tendency to defend and justify the societal status quo in the face of the threat posed by environmental problems. The present research finds that system justification tendencies are associated with greater denial of environmental realities and less commitment to pro-environmental action. Moreover, the effects of political conservatism, national identification, and gender on denial of environmental problems are explained by variability in system justification tendencies. However, this research finds that it is possible to eliminate the negative effect of system justification on environmentalism by encouraging people to regard pro environmental change as patriotic and consistent with protecting the status quo (i.e., as a case of "system-sanctioned change"). Theoretical and practical implications of these findings are discussed. PMID- 20008966 TI - The "shoulds" and "should nots" of moral emotions: a self-regulatory perspective on shame and guilt. AB - A self-regulatory framework for distinguishing between shame and guilt was tested in three studies. Recently, two forms of moral regulation based on approach versus avoidance motivation have been proposed in the literature. Proscriptive regulation is sensitive to negative outcomes, inhibition based, and focused on what we should not do. Prescriptive regulation is sensitive to positive outcomes, activation based, and focused on what we should do. In the current research, consistent support was found for shame's proscriptive and guilt's prescriptive moral underpinnings. Study 1 found a positive association between avoidance orientation and shame proneness and between approach orientation and guilt proneness. In Study 2, priming a proscriptive orientation increased shame and priming a prescriptive orientation increased guilt. In Study 3, transgressions most apt to represent proscriptive and prescriptive violations predicted subsequent judgments of shame and guilt, respectively. This self-regulatory perspective provides a broad interpretive framework for understanding and extending past research findings. PMID- 20008967 TI - Regulatory accessibility and social influences on state self-control. AB - The current work examined how social factors influence self-control. Current conceptions of state self-control treat it largely as a function of regulatory capacity. The authors propose that state self-control might also be influenced by social factors because of regulatory accessibility. Studies 1 through 4 provide evidence that individuals' state self-control is influenced by the trait and state self-control of salient others such that thinking of others with good trait or state self-control leads to increases in state self-control and thinking of others with bad trait or state self-control leads to decreases in state self control. Study 5 provides evidence that the salience of significant others influences both regulatory accessibility and state self-control. Combined, these studies suggest that the effects of social influences on state self-control occur through multiple mechanisms. PMID- 20008968 TI - Auto-appendectomy in the Antarctic: case report. PMID- 20008969 TI - Energy drinks for children and adolescents. PMID- 20008972 TI - Secret remedies: 100 years on. PMID- 20008976 TI - Dutch law leads to confusion over when to use life ending treatment in suffering newborns. PMID- 20008977 TI - In vivo tumor grading of prostate cancer using quantitative 111In-capromab pendetide SPECT/CT. AB - We have developed an in vivo method to quantify antibody uptake using (111)In capromab pendetide SPECT combined with CT (SPECT/CT). Our goal was to evaluate this method for potential grading of prostate tumors. METHODS: Our phantom experiments focused on the robustness of an advanced iterative reconstruction algorithm that involves corrections for photon attenuation, scatter, and geometric blurring caused by radionuclide collimators. The conversion factors between image values and tracer concentrations (in Bq/mL) were calculated from a uniform phantom filled with an aqueous solution of (111)InCl(3) using the same acquisition protocol and reconstruction parameters as for patient studies. In addition, the spatial resolution of the reconstructed images was measured from a point source phantom. The measured spatial resolution was modeled into a point spread function, and the point-spread function was implemented in a deconvolution based partial-volume-error correction algorithm. The recovery capability to correctly estimate true tracer concentrations was tested using prostatelike and bladderlike lesion phantoms fitted in the modified National Electrical Manufacturers Association/International Electrotechnical Commission body phantom. Patients with biopsy-proven prostate cancer (n = 10) who underwent prostatectomy were prospectively enrolled in the preoperative SPECT/CT studies at the San Francisco Veterans Affairs Medical Center. The CT portion of SPECT/CT was used to generate CT-based attenuation maps and as an anatomic localization tool for clinical interpretation. Pathologic Gleason grades were compared with in vivo antibody uptake value (AUV) normalized by injected dose, effective half-life, and injection-scan time difference. AUVs were calculated in each lobe of the prostate gland with cylindric volumes of interest having dimensions of 1.5 cm in both diameter and height. RESULTS: Reconstructed SPECT images further corrected by the deconvolution-based partial-volume-error correction could recover tracer concentrations up to 90% of true values in measurements of phantom volumes as small as 7.77 mL. From patient studies, there was a statistically significant correlation (rho = 0.71, P = 0.033) between higher AUVs (from either left or right lobe) and higher components of pathologic Gleason scores. CONCLUSION: Our results strongly indicate potential for noninvasive prostate tumor grading using quantitative (111)In-capromab pendetide SPECT/CT. PMID- 20008978 TI - Molecular imaging of atherosclerotic plaque with (64)Cu-labeled natriuretic peptide and PET. AB - Cardiovascular disease is the leading cause of death worldwide. PET has the potential to provide information on the biology and metabolism of atherosclerotic plaques. Natriuretic peptides (NPs) have potent antiproliferative and antimigratory effects on vascular smooth-muscle cells (VSMCs) and, in atherosclerosis, participate in vascular remodeling, in which the expression of NP clearance receptors (NPR-Cs) is upregulated both in endothelium and in VSMCs. METHODS: We investigated the potential of a C-type atrial natriuretic factor (C ANF) to image developing plaque-like lesions in vivo. C-ANF was functionalized with 1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetic acid (DOTA) and labeled with (64)Cu for noninvasive PET in a hypercholesterolemic rabbit with atherosclerotic-like lesions induced by air desiccation of a femoral artery, followed by balloon overstretch of the developing neointima. Histopathology and immunohistochemistry were performed to assess plaque development and NPR-C localization. RESULTS: (64)Cu-DOTA-C-ANF uptake in the atherosclerotic region was visible on small-animal PET images, with the highest target-to-background ratio (3.59 +/- 0.94) observed after the air desiccation-induced injury. Immunohistochemistry and immunofluorescence staining showed NPR-C near the luminal surface of the plaque and in VSMCs. PET and immunohistochemistry competitive blocking studies confirmed receptor-mediated tracer uptake in the plaque. With blocking, PET tracer localization of atherosclerotic to control arteries was decreased from 1.42 +/- 0.02 to 1.06 +/- 0.06 (P < 0.001). CONCLUSION: We demonstrated that (64)Cu-DOTA-C-ANF is a promising candidate tracer for in vivo PET of NPR-Cs on atherosclerotic plaques. PMID- 20008980 TI - Biodistribution and radiation dosimetry in humans of a new PET ligand, (18)F PBR06, to image translocator protein (18 kDa). AB - As a PET biomarker for inflammation, translocator protein (18 kDa) (TSPO) can be measured with an (18)F-labeled aryloxyanilide, (18)F-N-fluoroacetyl-N-(2,5 dimethoxybenzyl)-2-phenoxyaniline ((18)F-PBR06), in the human brain. The objective of this study was to estimate the radiation absorbed doses of (18)F PBR06 based on biodistribution data in humans. METHODS: After the injection of (18)F-PBR06, images were acquired from head to thigh in 7 healthy humans. Urine was collected at various time points. Radiation absorbed doses were estimated by the MIRD scheme. RESULTS: Moderate to high levels of radioactivity were observed in organs with high densities of TSPO and in organs of metabolism and excretion. Bone had low levels of radioactivity. The effective dose was 18.5 muSv/MBq. CONCLUSION: The effective dose of (18)F-PBR06, compared with other (18)F radioligands, was moderate. This radioligand had negligible defluorination, as indirectly assessed by bone radioactivity. Doses to the gallbladder wall and spleen may limit the amount of permissible injected radioactivity. PMID- 20008981 TI - Choline PET for monitoring early tumor response to photodynamic therapy. AB - Photodynamic therapy (PDT) is a relatively new therapy that has shown promise for treating various cancers in both preclinical and clinical studies. The present study evaluated the potential use of PET with radiolabeled choline to monitor early tumor response to PDT in animal models. METHODS: Two human prostate cancer models (PC-3 and CWR22) were studied in athymic nude mice. A second-generation photosensitizer, phthalocyanine 4 (Pc 4), was delivered to each animal by a tail vein injection 48 h before laser illumination. Small-animal PET images with (11)C choline were acquired before PDT and at 1, 24, and 48 h after PDT. Time-activity curves of (11)C-choline uptake were analyzed before and after PDT. The percentage of the injected dose per gram of tissue was quantified for both treated and control tumors at each time point. In addition, Pc 4-PDT was performed in cell cultures. Cell viability and (11)C-choline uptake in PDT-treated and control cells were measured. RESULTS: For treated tumors, normalized (11)C-choline uptake decreased significantly 24 and 48 h after PDT, compared with the same tumors before PDT (P < 0.001). For the control tumors, normalized (11)C-choline uptake increased significantly. For mice with CWR22 tumors, the prostate-specific antigen level decreased 24 and 48 h after PDT. Pc 4-PDT in cell culture showed that the treated tumor cells, compared with the control cells, had less than 50% (11)C-choline activity at 5, 30, and 45 min after PDT, whereas the cell viability test showed that the treated cells were viable longer than 7 h after PDT. CONCLUSION: PET with (11)C-choline is sensitive for detecting early changes associated with Pc 4-PDT in mouse models of human prostate cancer. Choline PET has the potential to determine whether a PDT-treated tumor responds to treatment within 48 h after therapy. PMID- 20008982 TI - Pharmacokinetic analysis of hypoxia (18)F-fluoromisonidazole dynamic PET in head and neck cancer. AB - This study used pharmacokinetic analysis of (18)F-labeled fluoromisonidazole ((18)F-FMISO) dynamic PET to assist the identification of regional tumor hypoxia and to investigate the relationship among a potential tumor hypoxia index (K(i)), tumor-to-blood ratio (T/B) in the late-time image, plasma-to-tissue transport rate (k(1)), and local vascular volume fraction (beta) for head and neck cancer patients. METHODS: Newly diagnosed patients underwent a dynamic (18)F-FMISO PET scan before chemotherapy or radiotherapy. The data were acquired in 3 consecutive PET/CT dynamic scan segments, registered with each other and analyzed using pharmacokinetics software. The (K(i), k(1), beta) kinetic parameter images were derived for each patient. RESULTS: Nine patients' data were analyzed. Representative images of (18)F-FDG PET (of the tumor), CT (of the anatomy), and late-time (18)F-FMISO PET (of the T/B) and parametric images of K(i) (potentially representing tumor hypoxia) are shown. The patient image data could be classified into 3 types: with good concordance between the parametric hypoxia map K(i) and high T/B, with concordant findings between the parametric hypoxia map and low T/B, and with ambiguity between parametric hypoxia map and T/B. Correlation coefficients are computed between each pair of T/B, K(i), k(1), and beta. Data are also presented for other potential hypoxia surrogate measures, for example, k(3) and k(1)/k(2). CONCLUSION: There is a positive correlation of 0.86 between the average T/B and average hypoxia index K(i) of the region of interest. However, because of the statistical photon counting noise in PET and the amplification of noise in kinetic analysis, the direct correlation between the T/B and hypoxia of the individual pixel is not obvious. For a tumor region of interest, there is a slight negative correlation between k(1) and K(i), moderate positive correlation between beta and K(i), but no correlation between beta and k(1). PMID- 20008983 TI - A novel monoclonal antibody for RAGE-directed imaging identifies accelerated atherosclerosis in diabetes. AB - Receptor for advanced glycation end products (RAGE) binds advanced glycation end products and other inflammatory ligands and is expressed in atherosclerotic plaques in diabetic and nondiabetic subjects. The higher expression in diabetes mellitus corresponds to the accelerated course of the atherosclerosis. This study was designed to test the hypothesis that the level of RAGE expression in atherosclerosis can be detected by quantitative in vivo SPECT and that counts in the target will correlate with the strength of the biologic signal. METHODS: A monoclonal murine antibody was developed against the V-domain of RAGE, fragmented into F(ab')(2) and labeled with (99m)Tc, and injected at a dose of 15.14 +/- 1.23 MBq into 24-wk-old male apolipoprotein E null (ApoE(-/-)) mice (n = 22), including mice with streptozotocin-induced diabetes mellitus (n = 8), nondiabetic mice (n = 8), and control ApoE(-/-)/RAGE(-/-) double-knock-out mice (n = 6). Four hours later (allowing for blood-pool clearance), the mice were imaged and sacrificed, and the proximal aorta was removed and counted to calculate the percentage injected dose of RAGE per gram of tissue, followed by histologic and immunohistochemical characterization. RESULTS: Radiotracer uptake in the aortic lesions was clearly visualized noninvasively by SPECT. RAGE uptake as percentage injected dose in diabetic ApoE(-/-) mice (1.39 +/- 0.16 x 10(-2)) was significantly higher than that in nondiabetic ApoE(-/-) mice (0.48 +/- 0.27 x 10( 2)) (P < 0.0001). The radiotracer uptake was highly correlated with RAGE expression by quantitative immunohistomorphometry (r = 0.82, P = 0.002) and with percentage of macrophages (r = 0.86, P < 0.0001). CONCLUSION: In this study, (99m)Tc-labeled anti-RAGE F(ab')(2) SPECT successfully identified early accelerated disease in diabetes mellitus for age-matched ApoE(-/-) mice and quantified RAGE expression over a range of lesion severities. PMID- 20008984 TI - Combined (18)F-FDG and fluoride approach in PET/CT imaging: is there a clinical future? PMID- 20008985 TI - Imaging spontaneous MMTVneu transgenic murine mammary tumors: targeting metabolic activity versus genetic products. AB - Despite the great strides made in imaging breast cancer (BC) in humans, the current imaging modalities miss up to 30% of BC, do not distinguish malignant lesions from benign ones, and require histologic examinations for which invasive biopsy must be performed. Annually in the United States, approximately 5.6 million biopsies find benign lesions. More than 50% of human BCs overexpress cyclin D1, and all BCs exhibit VPAC1 oncogene products. Together, these gene products may provide an excellent biomarker for the early and accurate detection of BC. We have evaluated 4 biologically active peptide analogs that have high affinity for VPAC1. The transgenic MMTVneu mice spontaneously develop BC and metastatic lesions that overexpress cyclin D1 and VPAC1 biomarkers. The MMTVneu mouse, therefore, provides an excellent animal model that mimics the pathogenesis of human BC. The objective of this investigation was to determine the ability of 1 of the peptide analogs, (64)Cu-TP3805, to detect BC in MMTVneu mice using (18)F FDG as a gold standard. METHODS: The transgenic MMTVneu mouse colony was maintained. Offspring were screened for transgenic status by reverse transcriptase polymerase chain reaction (RT-PCR). Nine mice with visible, palpable, or unknown metastatic lesions were entered into the protocol. (18)F-FDG (6,475 +/- 1,628 kBq [175 +/- 44 microCi]) PET served as a control, followed by a CT scan and 24-48 h later by PET with (64)Cu-TP3805 (4,588 +/- 962 kBq [124 +/- 26 microCi]). RT-PCR on excised tumors determined VPAC1 expression, and histology ascertained the pathology. RESULTS: Ten tumors were detected by PET. Four tumors were detected both by (18)F-FDG and by (64)Cu-TP3805. Additionally, 4 tumors were imaged with (64)Cu-TP3805 only. These 8 tumors overexpressed VPAC1 receptors and were malignant by histology. The 2 remaining tumors were visualized with (18)F FDG only. These tumors did not express the VPAC1 oncogene product and had benign histology. The standard uptake value ranged from 3.1 to 18.3 for (64)Cu-TP3805 and 0.9 to 1.4 for (18)F-FDG. CONCLUSION: (64)Cu-TP3805 identified all malignant lesions unequivocally that overexpressed the VPAC1 oncogene surface product. The 2 benign tumors that did not express the VPAC1 receptor were not imaged. (64)Cu TP3805 promises to have the potential for the early and accurate imaging of primary and metastatic BC. PMID- 20008986 TI - A nucleolin-targeted multimodal nanoparticle imaging probe for tracking cancer cells using an aptamer. AB - The recent advances in molecular imaging techniques, using cancer-targeting nanoparticle probes, provide noninvasive tracking information on cancer cells in living subjects. Here, we report a multimodal cancer-targeted imaging system capable of concurrent fluorescence imaging, radionuclide imaging, and MRI in vivo. METHODS: A cobalt-ferrite nanoparticle surrounded by fluorescent rhodamine (designated MF) within a silica shell matrix was synthesized with the AS1411 aptamer (MF-AS1411) that targets nucleolin (a cellular membrane protein highly expressed in cancer) using N-(3-dimethylaminopropyl)-N-ethylcarbodiimide (EDC). This purified MF-AS1411 particle was bound with 2-(p-isothio-cyanatobenzyl)-1,4,7 triazacyclonane-1,4,7-triacetic acid (p-SCN-bn-NOTA) chelating agent and further labeled with (67)Ga-citrate (MFR-AS1411). The shape and size distribution of MFR AS1411 were characterized by transmission electron microscope (TEM). The cellular distribution of the nucleolin protein using the MFR-AS1411 nanoparticle was detected by fluorescence confocal microscopy. Phantom MR images were obtained as the concentration of MFR-AS1411 increased, using a 1.5-T MRI scanner. In vivo (67)Ga radionuclide imaging and MRI were performed using a gamma-camera and a 1.5 T MR imager, respectively. RESULTS: TEM imaging revealed MF and MFR-AS1411 to be spheric and well dispersed. The purified MFR-AS1411 nanoparticle showed specific fluorescence signals in nucleolin-expressing C6 cells, compared with MFR-AS1411 mutant (MFR-AS1411mt)-treated C6 cells. The rhodamine fluorescence intensity and (67)Ga activity of MFR-AS1411 were enhanced in a dose-dependent manner as the concentration of MFR-AS1411 was increased. The (67)Ga radionuclide was detected in both thighs of the mice injected with MFR-AS1411, whereas the MFR-AS1411 mutant (MFR-AS1411mt) administration revealed rapid clearance via the bloodstream, demonstrating that MFR-AS1411 specifically targeted cancer cells. Bioluminescence images in the C6 cells, stably expressing the luciferase gene, illustrated the in vivo distribution. T2-weighted MR images of the same mice injected with MFR-AS1411 showed dark T2 signals inside the tumor region, compared with the MRI signal of the tumor region injected with MFR-AS1411mt particles. CONCLUSION: We developed a nanoparticle-based cancer-specific imaging probe using the AS1411 aptamer in vivo and in vitro. This multimodal targeting imaging strategy, using a cancer-specific AS1411 aptamer, can be used as a versatile imaging tool for specific cancer diagnosis. PMID- 20008987 TI - Spatial heterogeneity of lung perfusion assessed with (13)N PET as a vascular biomarker in chronic obstructive pulmonary disease. AB - Although it is known that structural and functional changes in the pulmonary vasculature and parenchyma occur in the progress of chronic obstructive pulmonary disease (COPD), information is limited on early regional perfusion (Q(r)) alterations. METHODS: We studied 6 patients with mild or moderate COPD and 9 healthy subjects (6 young and 3 age-matched). The PET (13)NN-labeled saline injection method was used to compute images of Q(r) and regional ventilation (V(r)). Transmission scans were used to assess regional density. We used the squared coefficient of variation to quantify Q(r) heterogeneity and length-scale analysis to quantify the contribution to total perfusion heterogeneity of regions ranging from less than 12 to more than 108 mm. RESULTS: Perfusion distribution in COPD subjects showed larger Q(r) heterogeneity, higher contribution from large length scales and lower contribution from small length scales, and larger heterogeneity of Q(r) normalized by tissue density than did healthy subjects. Dorsoventral gradients of V(r) were present in healthy subjects, with larger ventilation in dependent regions, whereas no gradient was present in COPD. Heterogeneity of ventilation-perfusion ratios was larger in COPD. CONCLUSION: Q(r) is significantly redistributed in COPD. Q(r) heterogeneity in COPD patients is greater than in healthy subjects, mainly because of the contribution of large lung regions and not because of changes in tissue density or V(r). The assessment of spatial heterogeneity of lung perfusion with (13)NN-saline PET may serve as a vascular biomarker in COPD. PMID- 20008988 TI - Imaging and quantitation of cannabinoid CB1 receptors in human and monkey brains using (18)F-labeled inverse agonist radioligands. AB - We recently demonstrated that (11)C-MePPEP, a PET ligand for CB(1) receptors, has such high uptake in the human brain that it can be imaged for 210 min and that receptor density can be quantified as distribution volume (V(T)) using the gold standard of compartmental modeling. However, (11)C-MePPEP had relatively poor retest and intersubject variabilities, which were likely caused by errors in the measurements of radioligand in plasma at low concentrations by 120 min. We sought to find an analog of (11)C-MePPEP that would provide more accurate plasma measurements. We evaluated several promising analogs in the monkey brain and chose the (18)F-di-deutero fluoromethoxy analog ((18)F-FMPEP-d(2)) to evaluate further in the human brain. METHODS: (11)C-FMePPEP, (18)F-FEPEP, (18)F-FMPEP, and (18)F-FMPEP-d(2) were studied in 5 monkeys with 10 PET scans. We calculated V(T) using compartmental modeling with serial measurements of unchanged parent radioligand in arterial plasma and radioactivity in the brain. Nonspecific binding was determined by administering a receptor-saturating dose of rimonabant, an inverse agonist at the CB(1) receptor. Nine healthy human subjects participated in 17 PET scans using (18)F-FMPEP-d(2), with 8 subjects having 2 PET scans to assess retest variability. To identify sources of error, we compared intersubject and retest variability of brain uptake, arterial plasma measurements, and V(T). RESULTS: (18)F-FMPEP-d(2) had high uptake in the monkey brain, with greater than 80% specific binding, and yielded less radioactivity uptake in bone than did (18)F-FMPEP. High brain uptake with (18)F-FMPEP-d(2) was also observed in humans, in whom V(T) was well identified within approximately 60 min. Retest variability of plasma measurements was good (16%); consequently, V(T) had a good retest variability (14%), intersubject variability (26%), and intraclass correlation coefficient (0.89). V(T) increased after 120 min, suggesting an accumulation of radiometabolites in the brain. Radioactivity accumulated in the skull throughout the entire scan but was thought to be an insignificant source of data contamination. CONCLUSION: Studies in monkeys facilitated our development and selection of (18)F-FMPEP-d(2), compared with (18)F-FMPEP, as a radioligand demonstrating high brain uptake, high percentage of specific binding, and reduced uptake in bone. Retest analysis in human subjects showed that (18)F-FMPEP-d(2) has greater precision and accuracy than (11)C MePPEP, allowing smaller sample sizes to detect a significant difference between groups. PMID- 20008989 TI - Impact of rituximab treatment on (90)Y-ibritumomab dosimetry for patients with non-Hodgkin lymphoma. AB - To determine whether the therapeutic effect of (90)Y-ibritumomab might be enhanced by a full course of rituximab followed by single dose of (90)Y ibritumomab, the trial included pre- and post-rituximab treatment imaging with (111)In-ibritumomab and blood pharmacokinetics. Comparison of the pre- and post rituximab imaging and blood data allowed for the assessment of impact of rituximab on (90)Y-ibritumomab dosimetry. METHODS: Seventeen patients with relapsed B cell non-Hodgkin lymphoma first received 250 mg/m(2) of rituximab plus 185 MBq of (111)In-ibritumomab for initial dosimetry evaluation. In weeks 2-4, patients received 3 weekly 375 mg/m(2) doses of rituximab. In week 6, patients received a 250 mg/m(2) dose of rituximab plus 185 MBq of (111)In-ibritumomab for a second dosimetry evaluation. Five sequential, whole-body gamma-camera images were acquired after the (111)In-ibritumomab injection. Calculated radiation doses were based on patient-specific organ masses. For each patient, paired comparison of radiation doses before and after rituximab treatment was performed. Paired comparison of residence times for spleen and tumor was also performed. RESULTS: Before rituximab treatment, the median radiation dose (mGy/MBq) was 0.48 (range, 0.24-0.86) for total body, 3.7 (range, 2.1-11.6) for liver, 6.1 (range, 1.8-17.8) for spleen, 3.3 (range, 2.0-4.7) for kidneys, 2.4 (range, 1.3-3.7) for heart wall, 1.1 (range, 0.4-2.3) for lungs, 0.79 (range, 0.32-1.22) for marrow from blood, and 18.1 (range, 4.7-98.9) for tumor. Paired comparisons were performed in 16 patients only because human antimurine antibody developed in 1 patient. The median change was 0.007 mGy/MBq for body, -0.14 mGy/MBq for liver, -0.31 mGy/MBq for kidneys, 0.38 mGy/MBq for heart wall, -0.17 mGy/MBq for lungs, and 0.046 mGy/MBq for marrow from blood. The median change in residence time was -0.92 h for spleen and -0.24 h for tumor. The changes were statistically insignificant for total body, liver, kidneys, lungs, and marrow from blood. The median residence times, or mGy/MBq if there were no volume changes, decreased 24% for spleen (P = 0.0005) and 28% for tumor (P = 0.005). The median radiation dose to heart wall increased 16%, which was statistically significant (P = 0.002). CONCLUSION: Changes in (90)Y-ibritumomab dosimetry after 4 wk of rituximab treatment were not significant for most organs, except for the heart wall. The reduction of spleen and tumor residence times is more likely to be due to the therapeutic effects of rituximab. PMID- 20008990 TI - PET-derived biodistribution and dosimetry of the benzodiazepine receptor-binding radioligand (11)C-(R)-PK11195 in children and adults. AB - The PET tracer (11)C-(R)-PK11195 (PK) is an antagonist of the peripheral-type benzodiazepine binding site and allows the noninvasive imaging of microglial activation seen in several neurologic disorders affecting the mature and developing brain. The objective of this study was to derive the biodistribution and in vivo radiation dose estimates of PK in children studied for brain inflammatory conditions and in healthy adults. METHODS: Twenty-two children (mean age +/- SD, 9.5 +/- 4 y; range, 4-17 y; 10 girls) who underwent dynamic PK PET for conditions involving brain inflammation were studied. Seven healthy adults (age, 27.4 +/- 7.5 y; range, 22-41 y; 3 women) were evaluated using the same protocol. Normal-organ time-activity curves and residence times were derived and absorbed doses then calculated using the OLINDA software. Two other healthy young adults (1 man, 1 woman) also underwent sequential whole-body PET using a PET/CT scanner to obtain corresponding CT images and PK pharmacokinetics. RESULTS: PK uptake was highest in the gallbladder and urinary bladder, followed by the liver, kidney, bone marrow, salivary gland, and heart wall, with minimal localization in all other organs including normal brain and lungs. PK was excreted through the hepatobiliary and renal systems. The average effective dose equivalent was 11.6 +/- 0.6 microSv/MBq (mean +/- SD) for young children (age, 4-7 y), 7.7 +/- 1.0 microSv/MBq for older children (age, 8-12 y), 5.3 +/- 0.5 muSv/MBq for adolescents (age, 13-17 y), and 4.6 +/- 2.7 microSv/MBq for adults. The gallbladder wall received the highest radiation dose in children younger than 12 y, whereas the urinary bladder wall received the highest dose in older children and adults. For an administered activity of 17 MBq/kg (0.45 mCi/kg), the effective dose equivalent was about 5 mSv or below for all age groups. CONCLUSION: At clinically practical administered activities, the radiation dose from (11)C-PK11195 in both children and adults is comparable to that from other clinical PET tracers and diagnostic radiopharmaceuticals in routine clinical use. PMID- 20008991 TI - Combined (18)F-FDG and fluoride approach in PET/CT imaging: is there a clinical future? PMID- 20008992 TI - Integrated software environment based on COMKAT for analyzing tracer pharmacokinetics with molecular imaging. AB - An integrated software package, Compartment Model Kinetic Analysis Tool (COMKAT), is presented in this report. METHODS: COMKAT is an open-source software package with many functions for incorporating pharmacokinetic analysis in molecular imaging research and has both command-line and graphical user interfaces. RESULTS: With COMKAT, users may load and display images, draw regions of interest, load input functions, select kinetic models from a predefined list, or create a novel model and perform parameter estimation, all without having to write any computer code. For image analysis, COMKAT image tool supports multiple image file formats, including the Digital Imaging and Communications in Medicine (DICOM) standard. Image contrast, zoom, reslicing, display color table, and frame summation can be adjusted in COMKAT image tool. It also displays and automatically registers images from 2 modalities. Parametric imaging capability is provided and can be combined with the distributed computing support to enhance computation speeds. For users without MATLAB licenses, a compiled, executable version of COMKAT is available, although it currently has only a subset of the full COMKAT capability. Both the compiled and the noncompiled versions of COMKAT are free for academic research use. Extensive documentation, examples, and COMKAT itself are available on its wiki-based Web site, http://comkat.case.edu. Users are encouraged to contribute, sharing their experience, examples, and extensions of COMKAT. CONCLUSION: With integrated functionality specifically designed for imaging and kinetic modeling analysis, COMKAT can be used as a software environment for molecular imaging and pharmacokinetic analysis. PMID- 20008993 TI - Implementation of an automated respiratory amplitude gating technique for PET/CT: clinical evaluation. AB - Amplitude gating techniques have recently been shown to be better at suppressing respiratory motion artifacts than phase gating. However, most commercial PET/CT scanners are equipped with phase gating capabilities only. The objective of this article was to propose and evaluate using patient studies an automated respiratory amplitude gating technique that could be implemented on current whole body PET/CT scanners. A primary design feature of the proposed technique is to automatically match the respiratory amplitude captured during the CT scan with a corresponding amplitude during the PET scan. METHODS: The proposed amplitude gating technique consists of a CT scan, followed by a list-mode PET scan. The CT scan was acquired while the patient's respiratory motion was recorded by a monitoring device that determined the respiratory motion amplitude captured during the CT scan. A program was designed to inject triggers into the PET list stream whenever the patient's respiration crossed a preset amplitude range determined by the captured amplitude during CT. To implement this proposed amplitude gating technique in whole-body PET/CT, a PET-first protocol was necessary to minimize the respiratory baseline drift between the CT and PET scans. In this implementation, a regular PET scan was first acquired over the patient's whole body but excluding the bed position that covered the lesion of interest. The whole-body CT scan was then acquired, followed by a list-mode PET acquisition over the bed position that covered the area of interest (lesion). The proposed amplitude gating technique was tested using 13 patients with 21 lung or thoracic tumors. RESULTS: In the patient studies, the gated images-when compared with the ungated images-showed statistically significant improvements, with an average 27% and 28% increase in maximum and mean standardized uptake value, respectively, for all lesions. Furthermore, the tumors in the gated images showed better contrast using visual inspection and line profiles. CONCLUSION: The implementation of the proposed respiratory amplitude gating technique on current PET/CT scanners is feasible, and amplitude-matched CT and PET data can be automatically generated using our proposed procedures without requiring patients to hold their breath or increase their radiation exposure. PMID- 20008994 TI - Diagnosis of cirrhotic portal hypertension and compensatory circulation using transsplenic portal scintigraphy with (99m)Tc-phytate. AB - Our objective was to investigate the diagnostic value and clinical significance of transsplenic portal scintigraphy in cirrhotic portal hypertension and compensatory circulation. METHODS: Transsplenic portal scintigraphy, ultrasound, and gastroscopy were performed on 50 patients with cirrhotic portal hypertension and on 10 controls. According to the Child-Pugh classification, 15 patients with cirrhosis were Child A, 19 were Child B, and 16 were Child C. RESULTS: In the control group, the splenoportal vein was shaped like the letter S, and the portosystemic shunt index was 0.19 +/- 0.07. Portal hypertension portosystemic shunts were of 3 types: intrahepatic (13 patients; index, 0.52 +/- 0.19), compensatory (31 patients; index, 0.64 +/- 0.28); and completely extrahepatic (6 patients; index, 0.91 +/- 0.03). Collateral vessels were uphill, downhill, or complex. The portosystemic shunt index increased as cirrhosis and esophageal varices increased. There was statistical significance among groups (P < 0.05 or < 0.01). CONCLUSION: Transsplenic portal scintigraphy was sensitive for detecting the number and location of shunts and will allow for improved surgical planning. PMID- 20008995 TI - Radioisotopic imaging of neuroinflammation. AB - Inflammatory responses are closely associated with many neurologic disorders and influence their outcome. In vivo imaging can document events accompanying neuroinflammation, such as changes in blood flow, vascular permeability, tightness of the blood-to-brain barrier, local metabolic activity, and expression of specific molecular targets. Here, we briefly review current methods for imaging neuroinflammation, with special emphasis on nuclear imaging techniques. PMID- 20008996 TI - The PET/CT report: the most important part of the study. PMID- 20008997 TI - PET and PET/CT reports: observations from the National Oncologic PET Registry. AB - Our objective was to identify core elements for inclusion in oncologic PET reports and to evaluate a sample of reports in the National Oncologic PET Registry database. METHODS: A list of desirable elements in PET reports was compiled from American College of Radiology and Society of Nuclear Medicine guidelines. A training set of 20 randomly selected reports was evaluated by the 4 physician panel, and the results were used to formulate a consensus approach for assessing report content and quality. Each reviewer then scored 65 randomly selected reports-20 common to all reviewers. The scores were tabulated, and interrater variability was measured for the common cases. RESULTS: Each report was assessed for 34 elements-21 primary and 11 additional questions related to 6 of these primary elements. Among the common cases, there was strong (> or = 0.70) interrater agreement for 30 of 34 elements. Among the unique cases, only 9 elements were included in more than 90% of the reports. Several important elements were not included in more than 40% of the reports: the reason for the study, a description of treatment history, a statement about comparison to other imaging, and time from radiopharmaceutical injection to imaging. CONCLUSION: Essential elements that should be included in oncologic PET reports were missing from many reports. These deficiencies may render the reports less helpful to referring physicians, may lead to misdiagnoses, and may cause coding and billing errors. Interpreting physicians should audit their reports to ascertain that they include appropriate elements necessary for billing compliance and for effective communications with referring physicians. PMID- 20008998 TI - Dopamine cell implantation in Parkinson's disease: long-term clinical and (18)F FDOPA PET outcomes. AB - We have previously reported the results of a 1-y double-blind, placebo-controlled study of embryonic dopamine cell implantation for Parkinson's disease. At the end of the blinded phase, we found a significant increase in putamen uptake on (18)F fluorodopa ((18)F-FDOPA) PET reflecting the viability of the grafts. Nonetheless, clinical improvement was significant only in younger (age < or = 60 y) transplant recipients, as indicated by a reduction in Unified Parkinson's Disease Rating Scale (UPDRS) motor scores. METHODS: We now report long-term clinical and PET outcomes from 33 of the original trial participants who were followed for 2 y after transplantation and 15 of these subjects who were followed for 2 additional years. Longitudinal changes in UPDRS motor ratings and caudate and putamen (18)F FDOPA uptake were assessed with repeated-measures ANOVA. Relationships between these changes over time were evaluated by the analysis of within-subject correlations. RESULTS: We found that UPDRS motor ratings declined over time after transplantation (P < 0.001). Clinical improvement at 1 y was relatively better for the younger transplant recipients and for men, but these age and sex differences were not evident at longer-term follow-up. Significant increases in putamen (18)F-FDOPA uptake were evident at all posttransplantation time points (P < 0.001) and were not influenced by either age or sex. Posttransplantation changes in putamen PET signal and clinical outcome were significantly intercorrelated (P < 0.02) over the course of the study. Image analysis at the voxel level revealed significant bilateral increases in (18)F-FDOPA uptake at 1 y (P < 0.001) in the posterior putamen engraftment sites. PET signal in this region increased further at 2 and 4 y after engraftment. Concurrently, this analysis disclosed progressive declines in radiotracer uptake in the nonengrafted caudate and ventrorostral putamen. Clinical improvement after transplantation correlated with the retention of PET signal in this region at the preoperative baseline. CONCLUSION: These results suggest that clinical benefit and graft viability are sustained up to 4 y after transplantation. Moreover, the dependence of clinical (but not imaging) outcomes on subject age and sex at 1 y may not persist over the long term. Last, the imaging changes reliably correlate with clinical outcome over the entire posttransplantation time course. PMID- 20008999 TI - Nuclear myocardial perfusion imaging with a cadmium-zinc-telluride detector technique: optimized protocol for scan time reduction. AB - We aimed at establishing the optimal scan time for nuclear myocardial perfusion imaging (MPI) on an ultrafast cardiac gamma-camera using a novel cadmium-zinc telluride (CZT) solid-state detector technology. METHODS: Twenty patients (17 male; BMI range, 21.7-35.5 kg/m(2)) underwent 1-d (99m)Tc-tetrofosmin adenosine stress and rest MPI protocols, each with a 15-min acquisition on a standard dual detector SPECT camera. All scans were immediately repeated on an ultrafast CZT camera over a 6-min acquisition time and reconstructed from list-mode raw data to obtain scan durations of 1 min, 2 min, etc., up to a maximum of 6 min. For each of the scan durations, the segmental tracer uptake value (percentage of maximum myocardial uptake) from the CZT camera was compared by intraclass correlation with standard SPECT camera data using a 20-segment model, and clinical agreement was assessed per coronary territory. Scan durations above which no further relevant improvement in uptake correlation was found were defined as minimal required scan times, for which Bland-Altman limits of agreement were calculated. RESULTS: Minimal required scan times were 3 min for low dose (r = 0.81; P < 0.001; Bland-Altman, -11.4% to 12.2%) and 2 min for high dose (r = 0.80; P < 0.001; Bland-Altman, -7.6% to 12.9%), yielding a clinical agreement of 95% and 97%, respectively. CONCLUSION: We have established the minimal scan time for a CZT solid-state detector system, which allows 1-d stress/rest MPI with a substantially reduced acquisition time resulting in excellent agreement with regard to uptake and clinical findings, compared with MPI from a standard dual head SPECT gamma-camera. PMID- 20009000 TI - Innovations in radiotherapy planning of head and neck cancers: role of PET. AB - Modern radiotherapy techniques heavily rely on high-quality medical imaging. PET provides biologic information about the tumor, complementary to anatomic imaging. Integrated PET/CT has found its way into the practice of radiation oncology, and (18)F-FDG PET is being introduced for radiotherapy planning. The functional information possibly augments accurate delineation and treatment of the tumor and its extensions while reducing the dose to surrounding healthy tissues. In addition to (18)F-FDG, other PET tracers are available for imaging specific biologic tumor characteristics determining radiation resistance. For head and neck cancer, the potential gains of PET are increasingly being recognized. This review describes the current role of PET and perspectives on its future use for selection and delineation of radiotherapy target volumes and for biologic characterization of this tumor entity. Furthermore, the potential role of PET for early response monitoring, treatment modification, and patient selection is addressed in this review. PMID- 20009002 TI - (18)F-FDG avidity in lymphoma readdressed: a study of 766 patients. AB - PET/CT with (18)F-FDG is an important noninvasive diagnostic tool for management of patients with lymphoma, and its use may surpass current guideline recommendations. The aim of the present study is to enlarge the growing body of evidence concerning (18)F-FDG avidity of lymphoma to provide a basis for future guidelines. METHODS: The reports from (18)F-FDG PET/CT studies performed in a single center for staging of 1,093 patients with newly diagnosed Hodgkin disease and non-Hodgkin lymphoma between 2001 and 2008 were reviewed for the presence of (18)F-FDG avidity. Of these patients, 766 patients with a histopathologic diagnosis verified according to the World Health Organization classification were included in the final analysis. (18)F-FDG avidity was defined as the presence of at least 1 focus of (18)F-FDG uptake reported as a disease site. Nonavidity was defined as disease proven by clinical examination, conventional imaging modalities, and histopathology with no (18)F-FDG uptake in any of the involved sites. RESULTS: At least one (18)F-FDG-avid lymphoma site was reported for 718 patient studies (94%). Forty-eight patients (6%) had lymphoma not avid for (18)F FDG. (18)F-FDG avidity was found in all patients (100%) with Hodgkin disease (n = 233), Burkitt lymphoma (n = 18), mantle cell lymphoma (n = 14), nodal marginal zone lymphoma (n = 8), and lymphoblastic lymphoma (n = 6). An (18)F-FDG avidity of 97% was found in patients with diffuse large B-cell lymphoma (216/222), 95% for follicular lymphoma (133/140), 85% for T-cell lymphoma (34/40), 83% for small lymphocytic lymphoma (24/29), and 55% for extranodal marginal zone lymphoma (29/53). CONCLUSION: The present study indicated that with the exception of extranodal marginal zone lymphoma and small lymphocytic lymphoma, most lymphoma subtypes have high (18)F-FDG avidity. The cumulating evidence consistently showing high (18)F-FDG avidity in the potentially curable Burkitt, natural killer/T-cell, and anaplastic large T-cell lymphoma subtypes justifies further investigations of the utility of (18)F-FDG PET in these diseases at presentation. PMID- 20009003 TI - Plerixafor for stem cell mobilization in patients with non-Hodgkin's lymphoma and multiple myeloma. AB - OBJECTIVE: To evaluate the literature characterizing the mechanism of action, pharmacokinetics, pharmacodynamics, and therapeutic efficacy of plerixafor for hematopoietic stem cell (HSC) mobilization for autologous transplantation in patients with non-Hodgkin's lymphoma (NHL) or multiple myeloma. DATA SOURCES: A PubMed search (1966-September 2009) was conducted using the key words plerixafor and AMD3100. Manufacturer's prescribing information was also used. STUDY SELECTION AND DATA EXTRACTION: English-language articles were selected and data were extracted with a focus on clinical studies of HSC mobilization in patients with NHL or multiple myeloma. DATA SYNTHESIS: Plerixafor exerts its effect by reversibly blocking the ability of HSC to bind to the bone marrow matrix. When used with granulocyte colony-stimulating factor (G-CSF), plerixafor helps increase the number of HSCs in the peripheral blood, where they can be collected for use in autologous transplantation. In clinical studies, plerixafor was rapidly absorbed after subcutaneous injection, reaching a maximum plasma concentration at approximately 0.5 hours. Plerixafor is renally excreted as the parent drug, with an elimination half-life ranging from 3 to 5 hours. Plerixafor increases circulating CD34+ cells in the peripheral blood, with a peak effect about 6-9 hours after subcutaneous administration. An approximate 2- to 3-fold increase in the CD34+ cell count is seen by the first dose of plerixafor after 4 consecutive days of G-CSF treatment. In 2 Phase 3 studies in patients with NHL or multiple myeloma, addition of plerixafor to G-CSF resulted in a higher CD34+ cell collection with fewer apheresis days, but failed to show better graft durability or overall patient survival for up to 12 months of follow-up. CONCLUSIONS: Clinical trials have demonstrated that the addition of plerixafor to G-CSF was beneficial for HSC mobilization to peripheral blood for collection and subsequent transplantation in patients with NHL or multiple myeloma. Further studies should assess the benefit of the additive use of plerixafor on clinical outcomes. PMID- 20009004 TI - Clopidogrel hypersensitivity: a novel multi-day outpatient oral desensitization regimen. AB - BACKGROUND: Clopidogrel hypersensitivity has posed a problem for the acute treatment and long-term care of a particular patient population with coronary artery disease and stent placement. Patients with clopidogrel hypersensitivity have had an increased risk of hypersensitivity reactions, including anaphylaxis, if they ingest clopidogrel without undergoing an oral desensitization procedure. The previously published desensitization protocols have either been performed in the intensive care unit, requiring significant cost and healthcare utilization, or have required a full-day outpatient commitment on behalf of the patient. OBJECTIVE: To determine whether a multi-day outpatient oral clopidogrel desensitization protocol is effective and safe for patients with clopidogrel hypersensitivity. METHODS: We retrospectively assessed the efficacy of a 10-dose outpatient multi-day clopidogrel desensitization protocol performed in a university allergy-immunology center from April 2006 to October 2008 in patients with clopidogrel hypersensitivity. Patients were desensitized over 2-3 half-day clinical visits and were able to go home between desensitization sessions. A preliminary cost analysis was performed using the average of actual costs for the outpatient clopidogrel desensitization procedure and was compared with the average cost for an inpatient oral desensitization completed at our institution. RESULTS: Eight patients with coronary artery disease, cardiac stent placement, and clopidogrel hypersensitivity underwent an outpatient multi-day oral clopidogrel desensitization procedure. All patients were successfully desensitized with the multi-day protocol without complications. No patient had recurrence of allergic reaction 3 months after the procedure. A preliminary cost analysis demonstrated a lower cost for the outpatient compared to the inpatient oral clopidogrel desensitization protocol. CONCLUSIONS: This outpatient 10-dose multi-day clopidogrel desensitization protocol is a safe and effective novel approach for the treatment of clopidogrel hypersensitivity in patients with coronary artery disease and cardiac stent placement. In addition to safety and efficacy, this protocol offers the patient the convenience of avoiding hospital admission or full-day time commitments. PMID- 20009005 TI - Cyclosporine versus tacrolimus in immunosuppressive maintenance regimens in renal transplants in Brazil: survival analysis from 2000 to 2004. AB - BACKGROUND: In Brazil, the National Health System (SUS) is responsible for almost all renal transplants. SUS protocols recommend using cyclosporine, in association with azathioprine and corticosteroids, to maintain the immunosuppression that is essential for successful renal transplant. Alternatively, cyclosporine can be replaced by tacrolimus. OBJECTIVE: To evaluate the effectiveness of therapeutic schema involving cyclosporine or tacrolimus after renal transplant during a 60 month follow-up period. METHODS: A historical cohort study, from 2000 to 2004, was conducted using 5686 patients who underwent renal transplant and received cyclosporine or tacrolimus. Uni - and multivariate analyses were performed using the Cox model to examine factors associated with progression to treatment failure. RESULTS: Most of the patients were male, aged 38 years or older, for whom the most frequent primary diagnosis of chronic renal failure (CRF) was glomerulonephritis/nephritis. Higher risk of treatment failure was associated with: therapeutic regimen (tacrolimus, HR 1.38, 95% CI 1.14 to 1.67), patient age at transplantation (additional year, HR 1.01, 95% CI 1.00 to 1.02), donor type (deceased, HR 1.60, 95% CI 1.35 to 1.89), median time of dialysis prior to transplantation (>24 mo, HR 1.29, 95% CI 1.09 to 1.52), and primary CRF diagnosis (diabetes, HR 1.54, 95% CI 1.09 to 2.17). CONCLUSIONS: The risk of treatment failure of patients receiving tacrolimus was observed to be 1.38 times that of those receiving cyclosporine, after adjusting the model for possible confounding factors such as patient sex, patient age, graft origin, prior time of dialysis, and cause of CRF. Our results were obtained from an observational study, and further studies are necessary to evaluate whether compliance with SUS clinical protocols could result in more effective care for renal transplant recipients. PMID- 20009006 TI - A clinical review of echinocandins in pediatric patients. AB - OBJECTIVE: To identify and evaluate available data on pediatric echinocandin use. DATA SOURCES: A PubMed search, limited to English-language articles, was conducted (1990-August 2009) using the search terms echinocandin, pediatric, child, pharmacokinetics, caspofungin, micafungin, and anidulafungin. Additional articles were retrieved from citations of selected references. STUDY SELECTION AND DATA EXTRACTION: Relevant information on the pharmacology, pharmacokinetics, efficacy, and safety of echinocandins in children was selected. Clinical trials, retrospective reviews, and case series were identified and evaluated. Data from these sources were included in this review. DATA SYNTHESIS: Caspofungin is the only echinocandin approved by the Food and Drug Administration for use in children. Pediatric pharmacokinetics has been evaluated with all 3 echinocandins but is limited with anidulafungin. Micafungin is the most well-studied agent in prospective clinical trials for antifungal prophylaxis in stem cell transplantation and treatment of invasive fungal infections. Caspofungin has been studied prospectively for febrile neutropenia and treatment of invasive fungal infections, but most published data are from retrospective reviews or case reports. One case report of anidulafungin for neonatal candidiasis has been published. The role of echinocandins in the management of invasive pediatric fungal infections has expanded. Micafungin and caspofungin are recommended as primary or alternative treatment of candidemia and esophageal or invasive candidiasis, and as salvage therapy for invasive aspergillosis. Micafungin is recommended for neutropenic prophylaxis in stem cell transplantation, while caspofungin may be used in febrile neutropenia as an alternative to azoles. Dosing has been well established for caspofungin only in children 3 months of age and above. Anidulafungin should be avoided in children until more pharmacokinetic and clinical data become available. CONCLUSIONS: Further comparative trials are needed to more clearly define the role of echinocandins, either as monotherapy or in combination for difficult-to-treat infections, in the pediatric population. PMID- 20009007 TI - Dynamic convergence and divergence of renal genomic and biological pathways in protection from Dahl salt-sensitive hypertension. AB - Chromosome 13 consomic and congenic rat strains were analyzed to investigate the pattern of genomic pathway utilization involved in protection against salt sensitive hypertension and renal injury. Introgression of the entire Brown-Norway chromosome 13 (consomic SS-13(BN)) or nonoverlapping segments of this chromosome (congenic strains, 16 Mbp in D13Rat151-D13Rat197 or 14 Mbp in D13Rat111-D13Got22) into the genome of the Dahl salt-sensitive rat attenuated salt-induced hypertension and proteinuria. mRNA abundance profiles in the renal cortex and the renal medulla from rats receiving 0.4% or 8% NaCl diets revealed two important features of pathway recruitment in these rat strains. First, the two congenic strains shared alterations in several pathways compared with Dahl salt-sensitive rats, despite the fact that the genomic segments introgressed in the two congenic strains did not overlap. Second, even though the genomic segment introgressed in each congenic strain was a part of the chromosome introgressed in the consomic strain, pathways altered in each congenic strain were not simply a subset of those altered in the consomic. Supporting the relevance of the mRNA data, differential expression of oxidative stress-related genes among the four strains of rats was associated with differences in urinary excretion of lipid peroxidation products. The findings suggest that different genetic alterations might converge to influence shared pathways in protection from hypertension, and that, depending on the genomic context, the same genetic alteration might diverge to affect different pathways. PMID- 20009008 TI - Mislocalization of mitochondria and compromised renal function and oxidative stress resistance in Drosophila SesB mutants. AB - Mitochondria accumulate at sites of intense metabolic activity within cells, but the adaptive value of this placement is not clear. In Drosophila, sesB encodes the ubiquitous isoform of adenine nucleotide translocase (ANT, the mitochondrial inner membrane ATP/ADP exchanger); null alleles are lethal, whereas hypomorphic alleles display sensitivity to a range of stressors. In the adult renal tubule, which is densely packed with mitochondria and hence enriched for sesB, both hypomorphic alleles and RNA interference knockdowns cause the mitochondria to lose their highly polarized distribution in the tissue and to become rounded. Basal cytoplasmic and mitochondrial calcium levels are both increased, and neuropeptide calcium response compromised, with concomitant defects in fluid secretion. The remaining mitochondria in sesB mutants are overactive and maintain depleted cellular ATP levels while generating higher levels of hydrogen peroxide than normal. When sesB expression is knocked down in just tubule principal cells, the survival of the whole organism upon oxidative stress is reduced, implying a limiting role for the tubule in homeostatic response to stressors. The physiological impacts of defective ANT expression are thus widespread and diverse. PMID- 20009009 TI - Transcriptional profiling reveals divergent roles of PPARalpha and PPARbeta/delta in regulation of gene expression in mouse liver. AB - Little is known about the role of the transcription factor peroxisome proliferator-activated receptor (PPAR) beta/delta in liver. Here we set out to better elucidate the function of PPARbeta/delta in liver by comparing the effect of PPARalpha and PPARbeta/delta deletion using whole genome transcriptional profiling and analysis of plasma and liver metabolites. In fed state, the number of genes altered by PPARalpha and PPARbeta/delta deletion was similar, whereas in fasted state the effect of PPARalpha deletion was much more pronounced, consistent with the pattern of gene expression of PPARalpha and PPARbeta/delta. Minor overlap was found between PPARalpha- and PPARbeta/delta-dependent gene regulation in liver. Pathways upregulated by PPARbeta/delta deletion were connected to innate immunity and inflammation. Pathways downregulated by PPARbeta/delta deletion included lipoprotein metabolism and various pathways related to glucose utilization, which correlated with elevated plasma glucose and triglycerides and reduced plasma cholesterol in PPARbeta/delta-/- mice. Downregulated genes that may underlie these metabolic alterations included Pklr, Fbp1, Apoa4, Vldlr, Lipg, and Pcsk9, which may represent novel PPARbeta/delta target genes. In contrast to PPARalpha-/- mice, no changes in plasma free fatty acid, plasma beta-hydroxybutyrate, liver triglycerides, and liver glycogen were observed in PPARbeta/delta-/- mice. Our data indicate that PPARbeta/delta governs glucose utilization and lipoprotein metabolism and has an important anti inflammatory role in liver. Overall, our analysis reveals divergent roles of PPARalpha and PPARbeta/delta in regulation of gene expression in mouse liver. PMID- 20009010 TI - Long human CHGA flanking chromosome 14 sequence required for optimal BAC transgenic "rescue" of disease phenotypes in the mouse Chga knockout. AB - Chromogranin A (CHGA) plays a catalytic role in formation of catecholamine storage vesicles and also serves as precursor to the peptide fragment catestatin, a catecholamine secretory inhibitor whose expression is diminished in the hypertensive individuals. We previously reported the hypertensive, hyperadrenergic phenotype of Chga-/- knockout (KO) mice and rescue by the human ortholog. In the present study, we compare two humanized CHGA mouse models. Into the Chga null background, by bacterial artificial chromosome transgenesis human CHGA transgene has been introduced. Both lines have the complete approximately 12 kbp CHGA gene integrated stably in the genome but have substantial differences in CHGA expression, as well as consequent sympathochromaffin biochemistry and physiology. A mouse model with longer-insert HumCHGA31 displays integration encompassing not only CHGA but also long human flanking sequences. This is in contrast to mouse model HumCHGA19 with limited flanking human sequence co integrated. As a consequence, HumCHGA19 mice have normal though diminished pattern of spatial expression of CHGA, and 14-fold lower circulating CHGA, with failure to rescue KO phenotypes to normalcy. In the longer-insert HumCHGA31 mice, catecholamine secretion, exaggerated responses to environmental stress, and hypertension were all alleviated. Promoter regions of the transgenes in both HumCHGA19 and HumCHGA31 display minimal CpG methylation, weighing against differential "position effects" of integration, and thus suggesting that lack of cis elements required for optimal CHGA expression occurs in HumCHGA19 mice. Such "humanized" CHGA mouse models may be useful in probing the physiological consequences of variation in CHGA expression found in humans, with consequences for susceptibility to hypertension and cardiovascular disease. PMID- 20009011 TI - Molecular markers of preterm labor in the choriodecidua. AB - Because relevant biochemical changes are known to begin at the choriodecidual interface some weeks before actual clinical onset of labor, we hypothesized that the preterm choriodecidua may display gene and protein expression patterns specific to preterm labor. Transcriptomic (microarray) and proteomic (2 dimensional gel electrophoresis [2DGE]) profiling methodologies were used to compare changes in choriodecidual tissue collected from women who delivered before 35 weeks of gestation following spontaneous preterm labor (n = 12) and gestation-matched nonlaboring controls (n = 7). Additionally, 2DGE was used to compare differences in protein expression during term and preterm labor and to construct a choriodecidual proteome map. Overall, expressed transcripts and proteins indicated active tissue remodeling independent of labor status and an association with inflammatory processes during labor. Spontaneous, infection induced and abruption-associated preterm deliveries were each defined by distinct transcriptional profiles. Proteins osteoglycin and progesterone receptor component 2 (PGRMC2) were upregulated during term and preterm labor while galectin 1, annexin 3, annexin 5, and protein disulfide isomerase (PDI) were upregulated only during preterm labor, suggesting a probable association with the underlying pathology. Together, these results represent novel data that warrant further investigations to elucidate plausible causal relationships of these molecules with spontaneous preterm delivery. PMID- 20009012 TI - Sex-specific and lineage-specific alternative splicing in primates. AB - Comparative studies of gene regulation suggest an important role for natural selection in shaping gene expression patterns within and between species. Most of these studies, however, estimated gene expression levels using microarray probes designed to hybridize to only a small proportion of each gene. Here, we used recently developed RNA sequencing protocols, which sidestep this limitation, to assess intra- and interspecies variation in gene regulatory processes in considerably more detail than was previously possible. Specifically, we used RNA seq to study transcript levels in humans, chimpanzees, and rhesus macaques, using liver RNA samples from three males and three females from each species. Our approach allowed us to identify a large number of genes whose expression levels likely evolve under natural selection in primates. These include a subset of genes with conserved sexually dimorphic expression patterns across the three species, which we found to be enriched for genes involved in lipid metabolism. Our data also suggest that while alternative splicing is tightly regulated within and between species, sex-specific and lineage-specific changes in the expression of different splice forms are also frequent. Intriguingly, among genes in which a change in exon usage occurred exclusively in the human lineage, we found an enrichment of genes involved in anatomical structure and morphogenesis, raising the possibility that differences in the regulation of alternative splicing have been an important force in human evolution. PMID- 20009013 TI - Introduction to "Behind the shield: a perspective on H1N1 from the inner terrain". PMID- 20009014 TI - The use of goal attainment scaling to facilitate and assess individualized change in a wellness intervention for women with fibromyalgia syndrome. AB - PURPOSE: The authors examined goal achievement over an 8-month period in women with fibromyalgia participating in a holistic health promotion intervention using Goal Attainment Scaling. DESIGN: Descriptive. METHODS: Goal attainment was assessed by interviews at five points: baseline (retrospectively), immediately after the 8-week classes, at the middle and end of the telephone support period, and 3 months later. FINDINGS: The percentage of women who attained or exceeded their goals in the four health promotion areas of Lifestyle Adjustment, Physical Activity, Nutrition, and Stress Management increased from between 15% and 26% at the end of the classes to between 58% and 76% at the end of the telephone support period. Although scores declined somewhat in the following 3 months, at least 60% of the women continued to report behaviors at or above their goal level in all areas except Physical Activity. CONCLUSIONS: Goal Attainment Scaling, an individually determined measure of change, can effectively capture behavioral changes associated with a holistic health promotion intervention. A nurse facilitator, who not only understood the need for specific, measurable GAS but was also able to encourage and support women's identification of goals that were meaningful to them, was key to the positive findings observed here. PMID- 20009015 TI - Therapeutic mirror interventions: an integrated review of the literature. AB - The mirror is an object that shows one a reflected image of body areas. There appears to be limited nursing literature about the role of a nurse in the use of a mirror. There is, however, literature of the therapeutic use of mirrors in medicine, neurology, psychiatry, psychology, physical medicine and rehabilitation, and rheumatology. The objective of this article is to review the basic elements and the underlying theoretical framework of mirror interventions. In 2007 to 2008, a keyword, abstract, and title search was conducted for therapeutic mirror studies between the years 1998 and 2008. A multidisciplinary, integrated review approach was used when it became apparent that there were limited nursing studies. Qualitative and quantitative strategies for reviewing evidence were used, and a narrative synthesis approach was used to guide the comprehensive synthesis. Underlying theoretical models were identified, and five elements of mirror interventions were synthesized from the literature. PMID- 20009017 TI - Identifying patterns in partnership with students who want to quit smoking. AB - Students who wanted to quit smoking were invited to partner with teachers/support persons to identify the meaningful patterns of their lives. The theoretical framework guiding the study was Newman's theory of health as expanding consciousness from the perspective of the unitary-transformative paradigm in nursing science. Student participants shared the meaningful relationships and events of their lives with their supportive partners, and together they reflected on the patterns of the students' lives. Two similarities among the individual patterns were found: a difficult experience in interpersonal relationships at some point in their lives and the initiation of smoking in their search for connectedness with friends. In the dialectic process, students experienced insight regarding their evolving patterns and made transformative changes in their lives, reflecting varying levels of tobacco control. PMID- 20009018 TI - Older people's experiences of dream coaching. AB - BACKGROUND: Recalling and talking about dreams could initiate dream work among older people and provide an opportunity for self-confrontation and personal growth, which could in turn promote gerotranscendental development. The present article describes older people's opinions about participating in a dream-coaching group; it also briefly describes the theoretical foundation of dream coaching. AIMS AND OBJECTIVES: The study aim was to investigate older people's experience of participating in a dream-coaching group based on Jungian psychology. DESIGN: A descriptive design was used. METHOD: Retrospective interviews were explored using qualitative content analysis. FINDINGS: The participants were satisfied with the arrangement of the dream-coaching groups. All participants believed that they had recalled their dreams and thought much more about their dreams during the period in which the dream-coaching group met. Three diverse appraisals of participating in a dream-coaching group, which had different effects on the participants, were identified: "An activity like any other activity," "An activity that led to deeper thoughts about the meaning of dreams," and "An activity that led to deeper thoughts both about the meaning of dreams and about how dreams can improve one's understanding of the life situation." CONCLUSIONS: It is possible to arrange dream-coaching groups for older people and could be a way to promote personal development using this type of intervention. IMPLICATIONS: The study provides some guidance as to how such a group could be organized, thus facilitating use of dream-coaching groups in gerontological care. PMID- 20009019 TI - Reflection of a 7-year patient care program: implementing and sustaining an integrative hospital program. AB - Integrative alternative therapies, also known as holistic therapies, have many applications in hospitals and health centers. These may include relaxation therapies, meditation, massage, reflexology, and Reiki or healing touch. Patients today are looking for these services, and institutions continue to explore ways to provide them without affecting their bottom line. The Integrative Cardiac Wellness Program is such a service, and its growth and longevity comes out of the personal investment of the staff to the program and to their patients. The literature review on the permanence of caring practice shows that caring about your work with patients, not just the job, is critical in longevity (Graber & Mitcham, 2004). The holistic nurses' and staff 's commitment to their professional growth within their specialty and their personal spiritual practice as experts in the field forms the backbone of the Integrative Wellness Program's success. It has been in existence for 7 years, providing integrative healing therapies to more than 7,000 patients, making it one of the most experienced. The program now serves cardiac surgery patients, and patients who have been diagnosed with cancer. PMID- 20009020 TI - Utilization of the natural helper model in health promotion targeting African American men. AB - When compared with other racial and ethnic groups, African American men experience a great number of health disparities. The factors that perpetuate health disparities among African American men are multidimensional and include lack of access to equitable health care, lack of knowledge and limited education about health promoting behaviors, lack of organizational trust and acceptability, the impact of masculinity on health, and psychological factors. This article explores the health promotion needs of African American men and the ability of the natural helper model to address those needs. Further discussion of its use by holistic nurses as a culturally tailored health promotion intervention targeting African American men is presented. PMID- 20009022 TI - Antagonistic HLH/bHLH transcription factors mediate brassinosteroid regulation of cell elongation and plant development in rice and Arabidopsis. AB - In rice (Oryza sativa), brassinosteroids (BRs) induce cell elongation at the adaxial side of the lamina joint to promote leaf bending. We identified a rice mutant (ili1-D) showing an increased lamina inclination phenotype similar to that caused by BR treatment. The ili1-D mutant overexpresses an HLH protein homologous to Arabidopsis thaliana Paclobutrazol Resistance1 (PRE1) and the human Inhibitor of DNA binding proteins. Overexpression and RNA interference suppression of ILI1 increase and reduce, respectively, rice laminar inclination, confirming a positive role of ILI1 in leaf bending. ILI1 and PRE1 interact with basic helix loop-helix (bHLH) protein IBH1 (ILI1 binding bHLH), whose overexpression causes erect leaf in rice and dwarfism in Arabidopsis. Overexpression of ILI1 or PRE1 increases cell elongation and suppresses dwarf phenotypes caused by overexpression of IBH1 in Arabidopsis. Thus, ILI1 and PRE1 may inactivate inhibitory bHLH transcription factors through heterodimerization. BR increases the RNA levels of ILI1 and PRE1 but represses IBH1 through the transcription factor BZR1. The spatial and temporal expression patterns support roles of ILI1 in laminar joint bending and PRE1/At IBH1 in the transition from growth of young organs to growth arrest. These results demonstrate a conserved mechanism of BR regulation of plant development through a pair of antagonizing HLH/bHLH transcription factors that act downstream of BZR1 in Arabidopsis and rice. PMID- 20009024 TI - Effects of fructosamine-3-kinase deficiency on function and survival of mouse pancreatic islets after prolonged culture in high glucose or ribose concentrations. AB - Due to their high glucose permeability, insulin-secreting pancreatic beta-cells likely undergo strong intracellular protein glycation at high glucose concentrations. They may, however, be partly protected from the glucotoxic alterations of their survival and function by fructosamine-3-kinase (FN3K), a ubiquitous enzyme that initiates deglycation of intracellular proteins. To test that hypothesis, we cultured pancreatic islets from Fn3k-knockout (Fn3k(-/-)) mice and their wild-type (WT) littermates for 1-3 wk in the presence of 10 or 30 mmol/l glucose (G10 or G30, respectively) and measured protein glycation, apoptosis, preproinsulin gene expression, and Ca(2+) and insulin secretory responses to acute glucose stimulation. The more potent glycating agent d-ribose (25 mmol/l) was used as positive control for protein glycation. In WT islets, a 1 wk culture in G30 significantly increased the amount of soluble intracellular protein-bound fructose-epsilon-lysines and the glucose sensitivity of beta-cells for changes in Ca(2+) and insulin secretion, whereas it decreased the islet insulin content. After 3 wk, culture in G30 also strongly decreased beta-cell glucose responsiveness and preproinsulin mRNA levels, whereas it increased islet cell apoptosis. Although protein-bound fructose-epsilon-lysines were more abundant in Fn3k(-/-) vs. WT islets, islet cell survival and function and their glucotoxic alterations were almost identical in both types of islets, except for a lower level of apoptosis in Fn3k(-/-) islets cultured for 3 wk in G30. In comparison, d-ribose (1 wk) similarly decreased preproinsulin expression and beta cell glucose responsiveness in both types of islets, whereas it increased apoptosis to a larger extent in Fn3k(-/-) vs. WT islets. We conclude that, despite its ability to reduce the glycation of intracellular islet proteins, FN3K is neither required for the maintenance of beta-cell survival and function under control conditions nor involved in protection against beta-cell glucotoxicity. The latter, therefore, occurs independently from the associated increase in the level of intracellular fructose-epsilon-lysines. PMID- 20009025 TI - Cellular modeling: insight into oral minimal models of insulin secretion. AB - The oral glucose tolerance test and meal tolerance test are common clinical tests of the glucose-insulin system. Several mathematical models have been suggested as means to extract information about beta-cell function from data from oral tolerance tests. Any such model needs to be fairly simple but should at the same time be linked to the underlying biology of the insulin-secreting beta-cells. The scope of the present work is to present a way to make such a connection using a recent model describing intracellular mechanisms. We show how the three main components of oral minimal secretion models, derivative control, proportional control, and delay, are related to subcellular events, thus providing mechanistic underpinning of the assumptions of the minimal models. PMID- 20009023 TI - PKA-dependent potentiation of glucose-stimulated insulin secretion by Epac activator 8-pCPT-2'-O-Me-cAMP-AM in human islets of Langerhans. AB - Potential insulin secretagogue properties of an acetoxymethyl ester of a cAMP analog (8-pCPT-2'-O-Me-cAMP-AM) that activates the guanine nucleotide exchange factors Epac1 and Epac2 were assessed using isolated human islets of Langerhans. RT-QPCR demonstrated that the predominant variant of Epac expressed in human islets was Epac2, although Epac1 was detectable. Under conditions of islet perifusion, 8-pCPT-2'-O-Me-cAMP-AM (10 microM) potentiated first- and second phase 10 mM glucose-stimulated insulin secretion (GSIS) while failing to influence insulin secretion measured in the presence of 3 mM glucose. The insulin secretagogue action of 8-pCPT-2'-O-Me-cAMP-AM was associated with depolarization and an increase of [Ca(2+)](i) that reflected both Ca(2+) influx and intracellular Ca(2+) mobilization in islet beta-cells. As expected for an Epac selective cAMP analog, 8-pCPT-2'-O-Me-cAMP-AM (10 microM) failed to stimulate phosphorylation of PKA substrates CREB and Kemptide in human islets. Furthermore, 8-pCPT-2'-O-Me-cAMP-AM (10 microM) had no significant ability to activate AKAR3, a PKA-regulated biosensor expressed in human islet cells by viral transduction. Unexpectedly, treatment of human islets with an inhibitor of PKA activity (H-89) or treatment with a cAMP antagonist that blocks PKA activation (Rp-8-CPT-cAMPS) nearly abolished the action of 8-pCPT-2'-O-Me-cAMP-AM to potentiate GSIS. It is concluded that there exists a permissive role for PKA activity in support of human islet insulin secretion that is both glucose dependent and Epac regulated. This permissive action of PKA may be operative at the insulin secretory granule recruitment, priming, and/or postpriming steps of Ca(2+)-dependent exocytosis. PMID- 20009026 TI - Skeletal muscle glucose uptake during contraction is regulated by nitric oxide and ROS independently of AMPK. AB - Reactive oxygen species (ROS) and nitric oxide (NO) have been implicated in the regulation of skeletal muscle glucose uptake during contraction, and there is evidence that they do so via interaction with AMP-activated protein kinase (AMPK). In this study, we tested the hypothesis that ROS and NO regulate skeletal muscle glucose uptake during contraction via an AMPK-independent mechanism. Isolated extensor digitorum longus (EDL) and soleus muscles from mice that expressed a muscle-specific kinase dead AMPKalpha2 isoform (AMPK-KD) and wild type litter mates (WT) were stimulated to contract, and glucose uptake was measured in the presence or absence of the antioxidant N-acetyl-l-cysteine (NAC) or the nitric oxide synthase (NOS) inhibitor N(G)-monomethyl-l-arginine (l-NMMA). Contraction increased AMPKalpha2 activity in WT but not AMPK-KD EDL muscles. However, contraction increased glucose uptake in the EDL and soleus muscles of AMPK-KD and WT mice to a similar extent. In EDL muscles, NAC and l-NMMA prevented contraction-stimulated increases in oxidant levels (dichloroflourescein fluorescence) and NOS activity, respectively, and attenuated contraction stimulated glucose uptake in both genotypes to a similar extent. In soleus muscles of AMPK-KD and WT mice, NAC prevented contraction-stimulated glucose uptake and l-NMMA had no effect. This is likely attributed to the relative lack of neuronal NOS in the soleus muscles compared with EDL muscles. Contraction increased AMPKalpha Thr(172) phosphorylation in EDL and soleus muscles of WT but not AMPK-KD mice, and this was not affected by NAC or l-NMMA treatment. In conclusion, ROS and NO are involved in regulating skeletal muscle glucose uptake during contraction via an AMPK-independent mechanism. PMID- 20009027 TI - Lack of functional GABA(B) receptors alters GnRH physiology and sexual dimorphic expression of GnRH and GAD-67 in the brain. AB - GABA, the main inhibitory neurotransmitter, acts through GABA(A/C) and GABA(B) receptors (GABA(B)Rs); it is critical for gonadotropin regulation. We studied whether the lack of functional GABA(B)Rs in GABA(B1) knockout (GABA(B1)KO) mice affected the gonadotropin axis physiology. Adult male and female GABA(B1)KO and wild-type (WT) mice were killed to collect blood and tissue samples. Gonadotropin releasing hormone (GnRH) content in whole hypothalami (HT), olfactory bulbs (OB), and frontoparietal cortexes (CT) were determined (RIA). GnRH expression by quantitative real-time PCR (qRT-PCR) was evaluated in preoptic area-anterior hypothalamus (POA-AH), medial basal-posterior hypothalamus (MBH-PH), OB, and CT. Pulsatile GnRH secretion from hypothalamic explants was measured by RIA. GABA, glutamate, and taurine contents in HT and CT were determined by HPLC. Glutamic acid decarboxylase-67 (GAD-67) mRNA was measured by qRT-PCR in POA-AH, MBH-PH, and CT. Gonadotropin content, serum levels, and secretion from adenohypophyseal cell cultures (ACC) were measured by RIA. GnRH mRNA expression was increased in POA-AH of WT males compared with females; this pattern of expression was inversed in GABA(B1)KO mice. MBH-PH, OB, and CT did not follow this pattern. In GABA(B1)KO females, GnRH pulse frequency was increased and GABA and glutamate contents were augmented. POA-AH GAD-67 mRNA showed the same expression pattern as GnRH mRNA in this area. Gonadotropin pituitary contents and serum levels showed no differences between genotypes. Increased basal LH secretion and decreased GnRH-stimulated gonadotropin response were observed in GABA(B1)KO female ACCs. These results support the hypothesis that the absence of functional GABA(B)Rs alters GnRH physiology and critically affects sexual dimorphic expression of GnRH and GAD-67 in POA-AH. PMID- 20009028 TI - RS4-type resistant starch prevents high-fat diet-induced obesity via increased hepatic fatty acid oxidation and decreased postprandial GIP in C57BL/6J mice. AB - Chemically modified starches (CMS) are RS4-type resistant starch, which shows a reduced availability, as well as high-amylose corn starch (HACS, RS2 type), compared with the corresponding unmodified starch. Previous studies have shown that RS4 increases fecal excretion of bile acids and reduces zinc and iron absorption in rats. The aim of this study was to investigate the effects of dietary RS4 supplementation on the development of diet-induced obesity in mice. Weight- and age-matched male C57BL/6J mice were fed for 24 wk on a high-fat diet containing unmodified starch, hydroxypropylated distarch phosphate (RS4), or HACS (RS2). Those fed the RS4 diet had significantly lower body weight and visceral fat weight than those fed either unmodified starch or the RS2 diet. Those fed the RS4 diet for 4 wk had a significantly higher hepatic fatty acid oxidation capacity and related gene expression and lower blood insulin than those fed either unmodified starch or the RS2 diet. Indirect calorimetry showed that the RS4 group exhibited higher energy expenditure and fat utilization compared with the RS2 group. When gavaged with fat (trioleate), RS4 stimulated a lower postprandial glucose-dependent insulinotropic polypeptide (GIP; incretin) response than RS2. Higher blood GIP levels induced by chronic GIP administration reduced fat utilization in high-fat diet-fed mice. In conclusion, dietary supplementation with RS4-type resistant starch attenuates high-fat diet-induced obesity more effectively than RS2 in C57BL/6J mice, which may be attributable to lower postprandial GIP and increased fat catabolism in the liver. PMID- 20009029 TI - Characterization of purinergic P2X4 receptor channels expressed in anterior pituitary cells. AB - Anterior pituitary cells express cation-conducting P2X receptor channels (P2XRs), but their molecular identity, electrophysiological properties, cell-specific expression pattern, and physiological roles have been only partially characterized. In this study, we show by quantitative RT-PCR that mRNA transcripts for the P2X(4) subunit are the most abundant in rat anterior pituitary tissue and confirm the P2X(4)R protein expression by Western blot analysis. Single-cell patch-clamp recordings show that extracellular ATP induced an inward depolarizing current in a majority of thyrotropin-releasing hormone responsive pituitary cells, which resembled the current profile generated by recombinant P2X(4)R. The channels were activated and desensitized in a dose dependent manner and deactivated rapidly. Activation of these channels led to stimulation of electrical activity and promotion of voltage-gated and voltage insensitive Ca(2+) influx. In the presence of ivermectin, a specific allosteric modulator of P2X(4)Rs, there was an approximately fourfold increase in the maximum amplitude of the ATP-induced inward current, accompanied by an increase in the sensitivity of receptors for ATP, slowed deactivation of receptors, and enhanced ATP-induced prolactin release. These results indicate that thyrotropin releasing hormone-responsive cells, including lactotrophs, express homomeric and/or heteromeric P2X(4)Rs, which facilitate Ca(2+) influx and hormone secretion. PMID- 20009030 TI - Overexpression of Foxf2 in adipose tissue is associated with lower levels of IRS1 and decreased glucose uptake in vivo. AB - Many members of the forkhead genes family of transcription factors have been implicated as important regulators of metabolism, in particular, glucose homeostasis, e.g., Foxo1, Foxa3, and Foxc2. The purpose of this study was to exploit the possibility that yet unknown members of this gene family play a role in regulating glucose tolerance in adipocytes. We identified Foxf2 in a screen for adipose-expressed forkhead genes. In vivo overexpression of Foxf2 in an adipose tissue-restricted fashion demonstrated that such mice display a significantly induced insulin secretion in response to an intravenous glucose load compared with wild-type littermates. In response to increased Foxf2 expression, insulin receptor substrate 1 (IRS1) mRNA and protein levels are significantly downregulated in adipocytes; however, the ratio of serine vs. tyrosine phosphorylation of IRS1 seems to remain unaffected. Furthermore, adipocytes overexpressing Foxf2 have a significantly lower insulin-mediated glucose uptake compared with wild-type adipocytes. These findings argue that Foxf2 is a previously unrecognized regulator of cellular and systemic whole body glucose tolerance, at least in part, due to lower levels of IRS1. Foxf2 and its downstream target genes can provide new insights with regard to identification of novel therapeutic targets. PMID- 20009031 TI - Glucose transporters in the 21st Century. AB - The ability to take up and metabolize glucose at the cellular level is a property shared by the vast majority of existing organisms. Most mammalian cells import glucose by a process of facilitative diffusion mediated by members of the Glut (SLC2A) family of membrane transport proteins. Fourteen Glut proteins are expressed in the human and they include transporters for substrates other than glucose, including fructose, myoinositol, and urate. The primary physiological substrates for at least half of the 14 Glut proteins are either uncertain or unknown. The well-established glucose transporter isoforms, Gluts 1-4, are known to have distinct regulatory and/or kinetic properties that reflect their specific roles in cellular and whole body glucose homeostasis. Separate review articles on many of the Glut proteins have recently appeared in this journal. Here, we provide a very brief summary of the known properties of the 14 Glut proteins and suggest some avenues of future investigation in this area. PMID- 20009032 TI - Early exposure of the pregestational intrauterine and postnatal growth-restricted female offspring to a peroxisome proliferator-activated receptor-{gamma} agonist. AB - Prenatal nutrient restriction with intrauterine growth restriction (IUGR) alters basal and glucose-stimulated insulin response and hepatic metabolic adaptation. The effect of early intervention with insulin-sensitizing peroxisome proliferator activated receptor gamma agonists was examined in the metabolically maladapted F(1) pregestational IUGR offspring with a propensity toward pregnancy-induced gestational diabetes. The effect of rosiglitazone maleate [RG; 11 micromol/day from postnatal day (PN) 21 to PN60] vs. placebo (PL) on metabolic adaptations in 2-mo-old F(1) female rats subjected to prenatal (IUGR), postnatal (PNGR), or pre- and postnatal (IUGR + PNGR) nutrient restriction was investigated compared with control (CON). RG vs. PL had no effect on body weight or plasma glucose concentrations but increased subcutaneous white and brown adipose tissue and plasma cholesterol concentrations in all three experimental groups. Glucose tolerance tests with a 1:1 mixture of [2-(2)H(2)]- and [6,6-(2)H(2)]glucose in RG IUGR vs. PL IUGR revealed glucose tolerance with a lower glucose-stimulated insulin release (GSIR) and suppressed endogenous hepatic glucose production (HGP) with no difference in glucose clearance (GC) and recycling (GR). RG PNGR, although similar to PL CON, was hyperglycemic vs. PL PNGR with reduced GR but no difference in the existent low GSIR, HGP, and GC. RG IUGR + PNGR overall was no different from the PL counterpart. Insulin tolerance tests revealed perturbed recovery to baseline from the exaggerated hypoglycemia in RG vs. the PL groups with the only exception being RG PNGR where further worsening of hypoglycemia over PL PNGR was minimal with full recovery to baseline. These observations support that early intervention with RG suppressed HGP in IUGR vs. PL IUGR, without increasing GSIR similar to that seen in CON. Although RG reversed PNGR to the PL CON metabolic state, no such insulin-sensitizing effect was realized in IUGR + PNGR. PMID- 20009033 TI - Multiple extrathymic precursors contribute to T-cell development with different kinetics. AB - T-cell development in the thymus depends on continuous supply of T-cell progenitors from bone marrow (BM). Several extrathymic candidate progenitors have been described that range from multipotent cells to lymphoid cell committed progenitors and even largely T-lineage committed precursors. However, the nature of precursors seeding the thymus under physiologic conditions has remained largely elusive and it is not known whether there is only one physiologic T-cell precursor population or many. Here, we used a competitive in vivo assay based on depletion rather than enrichment of classes of BM-derived precursor populations, thereby only minimally altering physiologic precursor ratios to assess the contribution of various extrathymic precursors to T-lineage differentiation. We found that under these conditions multiple precursors, belonging to both multipotent progenitor (MPP) and common lymphoid progenitor (CLP) subsets have robust T-lineage potential. However, differentiation kinetics of different precursors varied considerably, which might ensure continuous thymic output despite gated importation of extrathymic precursors. In conclusion, our data suggest that the thymus functions to impose T-cell fate on any precursor capable of filling the limited number of progenitor niches. PMID- 20009034 TI - LOX-1 as a natural IFN-alpha-mediated signal for apoptotic cell uptake and antigen presentation in dendritic cells. AB - The identification of molecules responsible for apoptotic cell (AC) uptake by dendritic cells (DCs) and induction of T-cell immunity against AC-associated antigens is a challenge in immunology. DCs differentiated in the presence of interferon-alpha (IFN-alpha-conditioned DCs) exhibit a marked phagocytic activity and a special attitude in inducing CD8(+) T-cell response. In this study, we found marked overexpression of the scavenger receptor oxidized low-density lipoprotein receptor 1 (LOX-1) in IFN-alpha-conditioned DCs, which was associated with increased levels of genes belonging to immune response families and high competence in inducing T-cell immunity against antigens derived from allogeneic apoptotic lymphocytes. In particular, the capture of ACs by IFN-alpha DCs led to a substantial subcellular rearrangement of major histocompatibility complex class I and class II molecules, along with enhanced cross-priming of autologous CD8(+) T cells and CD4(+) T-cell activation. Remarkably, AC uptake, CD8(+) T-cell cross priming, and, to a lesser extent, priming of CD4(+) T lymphocytes were inhibited by a neutralizing antibody to the scavenger receptor LOX-1 protein. These results unravel a novel LOX-1-dependent pathway by which IFN-alpha can, under both physiologic and pathologic conditions, render DCs fully competent for presenting AC-associated antigens for cross-priming CD8(+) effector T cells, concomitantly with CD4(+) T helper cell activation. PMID- 20009035 TI - Hypercholesterolemia promotes bone marrow cell mobilization by perturbing the SDF 1:CXCR4 axis. AB - Hypercholesterolemia is associated with elevated peripheral blood leukocytes and increased platelet levels, generally attributed to cholesterol-induced proinflammatory cytokines. Bone marrow (BM) cell mobilization and platelet production is achieved by disrupting the SDF-1:CXCR4 axis, namely with granulocyte colony-stimulating factor and/or CXCR4 antagonists. Here we show that high cholesterol disrupts the BM SDF-1:CXCR4 axis; promotes the mobilization of B cells, neutrophils, and progenitor cells (HPCs); and creates thrombocytosis. Hypercholesterolemia was achieved after a 30-day high-cholesterol feeding trial, resulting in elevated low-density lipoprotein (LDL) cholesterol levels and inversion of the LDL to high-density lipoprotein cholesterol ratio. Hypercholesterolemic mice displayed lymphocytosis, increased neutrophils, HPCs, and thrombocytosis with a lineage-specific decrease in the BM. Histologic analysis revealed that megakaryocyte numbers remained unaltered but, in high cholesterol mice, they formed large clusters in contact with BM vessels. In vitro, LDL induced stromal cell-derived factor-1 (SDF-1) production, suggesting that megakaryocyte delocalization resulted from an altered SDF-1 gradient. LDL also stimulated B cells and HPC migration toward SDF-1, which was blocked by scavenger receptor class B type I (cholesterol receptor) inhibition. Accordingly, hypercholesterolemic mice had increased peripheral blood SDF-1 levels, increased platelets, CXCR4-positive B lymphocytes, neutrophils, and HPCs. High cholesterol interferes with the BM SDF-1:CXCR4 axis, resulting in lymphocytosis, thrombocytosis, and HPC mobilization. PMID- 20009036 TI - Plasminogen activator inhibitor-1 (PAI-1) is cardioprotective in mice by maintaining microvascular integrity and cardiac architecture. AB - Although the involvement of plasminogen activator inhibitor-1 (PAI-1) in fibrotic diseases is well documented, its role in cardiac fibrosis remains controversial. The goal of this study was to determine the effect of a PAI-1 deficiency (PAI-1( /-)) on the spontaneous development of cardiac fibrosis. PAI-1(-/-) mice developed pervasive cardiac fibrosis spontaneously with aging, and these mice displayed progressively distorted cardiac architecture and markedly reduced cardiac function. To mechanistically elucidate the role of PAI-1 in cardiac fibrosis, 12-week-old mice were chosen to study the biologic events leading to fibrosis. Although fibrosis was not observed at this early age, PAI-1(-/-) hearts presented with enhanced inflammation, along with increased microvascular permeability and hemorrhage. A potent fibrogenic cytokine, transforming growth factor-beta (TGF-beta), was markedly enhanced in PAI-1(-/-) heart tissue. Furthermore, the expression levels of several relevant proteases associated with tissue remodeling were significantly enhanced in PAI-1(-/-) hearts. These results suggest that PAI-1 is cardioprotective, and functions in maintaining normal microvasculature integrity. Microvascular leakage in PAI-1(-/-) hearts may provoke inflammation, and predispose these mice to cardiac fibrosis. Therefore, a PAI-1 deficiency contributes to the development of cardiac fibrosis by increasing vascular permeability, exacerbating local inflammation, and increasing extracellular matrix remodeling, an environment conducive to accelerated fibrosis. PMID- 20009038 TI - Metabolic interactions among dietary cholesterol, copper, and fructose. PMID- 20009037 TI - How I treat influenza in patients with hematologic malignancies. AB - The 2009 H1N1 influenza pandemic has heightened the interest of clinicians for options in the prevention and management of influenza virus infection in immunocompromised patients. Even before the emergence of the novel 2009 H1N1 strain, influenza disease was a serious complication in patients with hematologic malignancies receiving chemotherapy or undergoing hematopoietic cell transplantation. Here we review the clinical manifestations of seasonal and 2009 H1N1 influenza and discuss current diagnosis, antiviral treatment, and prophylaxis options. We also summarize infection control and vaccination strategies for patients, family members, and caregivers. PMID- 20009039 TI - C. difficile outside hospital. PMID- 20009040 TI - Managing gastro-oesophageal reflux in infants. AB - Transient, inappropriate relaxation of the lower oesophageal sphincter may permit contents of the stomach to pass into the oesophagus (so-called gastro-oesophageal reflux [GOR]). This usually presents as regurgitation or vomiting and is common in infants, when it is usually mild and self-limiting, and requires no specific treatment. Gastro-oesophageal reflux disease (GORD) in infants describes reflux of gastric contents that causes troublesome symptoms or complications. GORD is sometimes wrongly diagnosed in healthy infants with troublesome but harmless symptoms of 'physiological' GOR. This has led to increasing, potentially inappropriate, use of acid reducing drugs. Furthermore, few of the drugs used to treat infants with GORD are licensed for this indication, a situation that DTB criticised 12 years ago. Here we consider GORD in infancy (i.e. in those aged 0 12 months), the treatments available and when these are needed. PMID- 20009041 TI - Strong opioids for osteoarthritis in primary care? AB - For some patients with osteoarthritis, commonly used drugs taken alone or in combination, offer insufficient pain relief, or are inappropriate. One issue this raises is whether strong opioids might then have a place in treatment. Some GPs may be reluctant to prescribe strong opioids for osteoarthritis. This may be due to concerns about addiction or long-term use of controlled drugs for non-cancer pain. Here we review the evidence on strong opioids as adjunctive treatment in osteoarthritis. PMID- 20009042 TI - Generic prescribing in epilepsy. AB - Most NHS prescriptions for medicines are written generically. However, there are controversies around generic prescribing in epilepsy, with many specialists believing that changing the supply of an antiepileptic risks reduced seizure control or unwanted effects. Here we review the evidence and other issues around generic prescribing of medicines used in epilepsy. PMID- 20009044 TI - Clinical effectiveness of implantable cardioverter-defibrillators among medicare beneficiaries with heart failure. AB - BACKGROUND: The clinical effectiveness of implantable cardioverter-defibrillators (ICDs) in older patients with heart failure has not been established, and older patients have been underrepresented in previous studies. METHODS AND RESULTS: We identified patients with heart failure who were aged 65 years or older and were eligible for an ICD, had left ventricular ejection fraction of 35% or less, and were discharged alive from hospitals participating in the Organized Program to Initiate Lifesaving Treatment in Hospitalized Patients With Heart Failure and the Get With the Guidelines-Heart Failure quality-improvement programs during the period January 1, 2003, through December 31, 2006. We matched the patients to Medicare claims to examine long-term outcomes. The main outcome measure was all cause mortality over 3 years. The study population included 4685 patients who were discharged alive and were eligible for an ICD. Mean age was 75.2 years, 60% of the patients were women, mean ejection fraction was 25%, and 376 (8.0%) patients received an ICD before discharge. Mortality was significantly lower among patients who received an ICD compared with those who did not (19.8% versus 27.6% at 1 year, 30.9% versus 41.9% at 2 years, and 38.1% versus 52.3% at 3 years; P<0.001 for all comparisons). The inverse probability-weighted adjusted hazard of mortality at 3 years for patients receiving an ICD was 0.71 (95% CI, 0.56 to 0.91). CONCLUSIONS: Medicare beneficiaries hospitalized with heart failure and left ventricular ejection fraction of 35% or less who were selected for ICD therapy had lower risk-adjusted long-term mortality compared with those who did not receive an ICD. Clinical Trial Registration- clinicaltrials.gov. Identifier: NCT00344513. PMID- 20009045 TI - The cover. Portrait of a child (winter). PMID- 20009046 TI - A piece of my mind. Jobs. PMID- 20009047 TI - More evidence on low vitamin D levels fuels push to revise recommended intake. PMID- 20009048 TI - Gene therapy repair of donor lungs improves outlook for transplantation. PMID- 20009049 TI - Aspirin use, colorectal cancer survival, and loss to follow-up. PMID- 20009050 TI - Population attributable risk of incident hypertension in women. PMID- 20009051 TI - Tobacco control in VA hospitals. PMID- 20009052 TI - Study design and palliative care intervention. PMID- 20009053 TI - Functional brain imaging in a woman with spatial neglect due to conversion disorder. PMID- 20009054 TI - Developers of the hepatitis B vaccine. PMID- 20009057 TI - Association between carrier screening and incidence of cystic fibrosis. AB - CONTEXT: A downward trend in cystic fibrosis (CF) birth incidence has been reported in some areas. OBJECTIVE: To evaluate the association between carrier screening and CF birth incidence. DESIGN, SETTING, AND PARTICIPANTS: In northeastern Italy, CF birth incidence is monitored by means of a long-standing neonatal screening program. In the same area, 2 sections using different carrier detection approaches were identified--the western region, in which CF carrier tests are offered only to relatives of patients or to couples planning in vitro fertilization; and the eastern region, in which carrier testing is offered to relatives and carrier screening to infertile couples and to couples of reproductive age. A total of 779,631 newborns underwent CF neonatal screening between January 1993 and December 2007, of whom 195 had CF detected. MAIN OUTCOME MEASURE: Cystic fibrosis birth incidence in the 2 regions. RESULTS: A time related decrease in birth incidence was found, with a mean annual percentage decrease of 0.16 per 10,000 neonates (P < .001). In the western region, 2559 carrier tests were performed, 314 carriers were identified, and 9 carrier couples were detected. In the eastern region, 87,025 carrier tests were performed, 3650 carriers were identified, and 82 carrier couples were detected. The birth rate decrease was greater in the eastern region (decrease rate, 0.24; 95% confidence interval [CI], 0.12-0.36) than in the western region (decrease rate, 0.04; 95% CI, -0.16 to 0.08; P = .01). The increase in the number of screened carriers over time was significantly correlated with the decrease in CF birth incidence (correlation coefficient = -0.53; 95% CI, -0.20 to -0.74; P = .003). CONCLUSION: In northeastern Italy, carrier screening was associated with a decrease in the incidence of CF. PMID- 20009056 TI - Association of plasma leptin levels with incident Alzheimer disease and MRI measures of brain aging. AB - CONTEXT: The adipokine leptin facilitates long-term potentiation and synaptic plasticity in the hippocampus, promotes beta-amyloid clearance, and improves memory function in animal models of aging and Alzheimer disease (AD). OBJECTIVE: To relate baseline circulating leptin concentrations in a community-based sample of individuals without dementia to incident dementia and AD during follow-up and magnetic resonance imaging (MRI) measures of brain aging in survivors. DESIGN, SETTING, AND PARTICIPANTS: Prospective study of plasma leptin concentrations measured in 785 persons without dementia (mean [SD] age, 79 [5] years; 62% female), who were in the Framingham original cohort at the 22nd examination cycle (1990-1994). A subsample of 198 dementia-free survivors underwent volumetric brain MRI between 1999 and 2005, approximately 7.7 years after leptin was assayed. Two measures of brain aging, total cerebral brain volume and temporal horn volume (which is inversely related to hippocampal volume) were assessed. MAIN OUTCOME MEASURE: Incidence of dementia and AD during follow-up until December 31, 2007. RESULTS: During a median follow-up of 8.3 years (range, 0-15.5 years), 111 participants developed incident dementia; 89 had AD. Higher leptin levels were associated with a lower risk of incident dementia and AD in multivariable models (hazard ratio per 1-SD increment in log leptin was 0.68 [95% confidence interval, 0.54-0.87] for all-cause dementia and 0.60 [95% confidence interval, 0.46-0.79] for AD). This corresponds to an absolute AD risk over a 12 year follow-up of 25% for persons in the lowest quartile (first quartile) vs 6% for persons in the fourth quartile of sex-specific leptin levels. In addition, a 1-SD elevation in plasma leptin level was associated with higher total cerebral brain volume and lower temporal horn volume, although the association of leptin level with temporal horn volume did not reach statistical significance. CONCLUSION: Circulating leptin was associated with a reduced incidence of dementia and AD and with cerebral brain volume in asymptomatic older adults. PMID- 20009058 TI - Heterogeneity is not always noise: lessons from improvement. AB - Rigorous experimental methods suppress differences among study participants (noise) to detect true intervention effects (signals). But suppressing participants' heterogeneity obscures an essential dimension of biological and clinical knowledge. Medicine is therefore ambivalent about the influence of heterogeneity on outcomes and struggles to find ways to take it properly into account in both clinical practice and research. This analysis explores the roots of that ambivalence. Drawing on the evaluation of 2 health care improvement initiatives, this article examines the unique features of improvement that help to understand heterogeneity's influence on study methods, and suggests a variety of ways to assess the effect of heterogeneity on study outcome measures. PMID- 20009055 TI - Effect of tarenflurbil on cognitive decline and activities of daily living in patients with mild Alzheimer disease: a randomized controlled trial. AB - CONTEXT: Amyloid-beta peptide (Abeta(42)) has been implicated in the pathogenesis of Alzheimer disease (AD). Tarenflurbil, a selective Abeta(42)-lowering agent, demonstrated encouraging results on cognitive and functional outcomes among mildly affected patients in an earlier phase 2 trial. OBJECTIVE: To determine the efficacy, safety, and tolerability of tarenflurbil. DESIGN, SETTING, AND PATIENTS: A multicenter, randomized, double-blind, placebo-controlled trial enrolling patients with mild AD was conducted at 133 trial sites in the United States between February 21, 2005, and April 30, 2008. Concomitant treatment with cholinesterase inhibitors or memantine was permitted. INTERVENTION: Tarenflurbil, 800 mg, or placebo, administered twice a day. MAIN OUTCOME MEASURES: Co-primary efficacy end points were the change from baseline to month 18 in total score on the subscale of the Alzheimer Disease Assessment Scale-Cognitive Subscale (ADAS Cog, 80-point version) and Alzheimer Disease Cooperative Studies-activities of daily living (ADCS-ADL) scale. Additional prespecified slope analyses explored the possibility of disease modification. RESULTS: Of the 1684 participants randomized, 1649 were included in the analysis, and 1046 completed the trial. Tarenflurbil had no beneficial effect on the co-primary outcomes (difference in change from baseline to month 18 vs placebo, based on least squares means: 0.1 for ADAS-Cog; 95% CI, -0.9 to 1.1; P = .86 and -0.5 for ADCS-ADL; 95% CI, -1.9 to 0.9; P = .48) using an intent-to-treat analysis. No significant differences occurred in the secondary outcomes. The ADAS-Cog score decreased by 7.1 points over 18 months. The tarenflurbil group had a small increase in frequency of dizziness, anemia, and infections. CONCLUSION: Tarenflurbil did not slow cognitive decline or the loss of activities of daily living in patients with mild AD. TRIAL REGISTRATION: clinicaltrials.gov Identifier: NCT00105547. PMID- 20009059 TI - Health insurance cooperatives: lessons from the Great Depression. PMID- 20009060 TI - The global breast cancer disparity: strategies for bridging the gap. PMID- 20009061 TI - Mentoring: what's in a name? PMID- 20009062 TI - Late-life dementias: does this unyielding global challenge require a broader view? PMID- 20009063 TI - Carrier screening, incidence of cystic fibrosis, and difficult decisions. PMID- 20009064 TI - JAMA patient page. Magnetic resonance imaging. PMID- 20009069 TI - Successful catheter ablation of a focal atrial tachycardia from the ascending aorta: a novel location and approach. PMID- 20009070 TI - Giant T-wave inversions and extreme QT prolongation. PMID- 20009071 TI - Letter by Brodsky regarding article "Treatment of atrial fibrillation with antiarrhythmic drugs or radiofrequency ablation: two systematic literature reviews and meta-analyses". PMID- 20009073 TI - Managing atrial fibrillation: catheter ablation or antiarrhythmic therapy? PMID- 20009074 TI - Remote magnetic catheter navigation for cavotricuspid isthmus ablation in patients with common-type atrial flutter. AB - BACKGROUND: Conventional catheter ablation of cavotricuspid isthmus (CTI) dependent atrial flutter is a widely applied standard therapy. Remote magnetic catheter navigation (RMN) may provide benefits for different ablation procedures, but its efficacy for CTI ablation has not been evaluated in a randomized, controlled trial. METHODS AND RESULTS: Ninety patients undergoing de novo ablation of atrial flutter were randomly assigned to conventional manual (n=45) or RMN-guided (n=45) CTI ablation with an 8-mm-tip catheter. Complete bidirectional isthmus block was achieved in 84% (RMN) and 91% (conventional catheter ablation) of the cases (P=0.52). RMN was associated with shorter fluoroscopy time (median, 10.6 minutes; interquartile range [IQR], 7.6 to 19.9, versus 15.0 minutes; IQR, 11.5 to 23.1; P=0.043) but longer total radiofrequency application (17.1 minutes; IQR, 8.6 to 25, versus 7.5 minutes; IQR, 3.6 to 10.9; P<0.0001), ablation time (55 minutes; IQR, 28 to 76, versus 17 minutes; IQR, 7 to 31; P<0.0001), and procedure duration (114+/-35 versus 77+/-24 minutes, P<0.0001). Procedure duration in the RMN group did not decrease significantly with case experience. Long-term procedure success, defined as achievement of complete CTI block and freedom from atrial flutter recurrence during 6 months of follow-up, was lower in the RMN group (73% versus 89%, P=0.063). Right atrial angiography after ablation revealed no significant differences between groups in terms of right atrial diameter or CTI length, morphology, and angulation. Furthermore, none of these parameters was predictive for difficult (ablation time >20 minutes) or unsuccessful ablation. CONCLUSIONS: RMN-guided CTI ablation is associated with reduced radiation exposure but prolonged ablation and procedure times as compared with conventional catheter navigation. Our findings suggest that ablation lesions produced with an RMN-guided 8-mm catheter are less effective irrespective of CTI anatomy. TRIAL REGISTRATION: clinicaltrials.gov Identifier: NCT00560872. PMID- 20009075 TI - Catheter ablation of atrial fibrillation without fluoroscopy using intracardiac echocardiography and electroanatomic mapping. AB - BACKGROUND: Catheter ablation of atrial fibrillation is currently guided by x-ray fluoroscopy. The associated radiation risk to patients and medical staff may be significant. We report an atrial fibrillation ablation technique using intracardiac echocardiography (ICE) and electroanatomic mapping without fluoroscopy. METHODS AND RESULTS: Twenty-one patients with atrial fibrillation (age, 42 to 73 years; 14 male; 14 paroxysmal, 7 persistent; body mass index, 26 to 38) underwent ablation. A decapolar catheter was advanced through the left subclavian vein until stable coronary sinus electrograms appeared on all electrodes. Two 9F sheaths were advanced transfemorally over a guide wire to the right atrium. A rotational ICE catheter was advanced through a deflectable sheath. Double transseptal puncture was performed with ICE guidance and facilitated by electrocautery. A 3D MRI left atrial image was registered to the ostia of the pulmonary veins using ICE. Catheter ablation was performed using ICE and electroanatomic mapping navigation. In 19 cases, no fluoroscopy was used and the staff did not wear protective lead. In 2 cases, 2 to 16 minutes of fluoroscopy was used to assist transseptal puncture. Median procedure time was 208 (188 to 221) minutes; coronary sinus cannulation took 5 (2 to 26) minutes; double transseptal took 26 (17 to 40) minutes; left atrial ablation time was 103 (90 to 127) minutes. All patients underwent circumferential pulmonary vein ablation and 8 patients underwent additional left atrial ablation. There were no procedure-related complications. CONCLUSIONS: Catheter ablation of atrial fibrillation without fluoroscopy is feasible and merits further attention. This technique may be especially helpful in preventing x-ray exposure in children, pregnant women, and obese patients undergoing left atrial ablation. PMID- 20009076 TI - Initial experience of assessing esophageal tissue injury and recovery using delayed-enhancement MRI after atrial fibrillation ablation. AB - BACKGROUND: Esophageal wall thermal injury after atrial fibrillation ablation is a potentially serious complication. However, no noninvasive modality has been used to describe and screen patients to examine whether esophageal wall injury has occurred. We describe a noninvasive method of using delayed-enhancement MRI to detect esophageal wall injury and subsequent recovery after atrial fibrillation ablation. METHODS AND RESULTS: We analyzed the delayed-enhancement MRI scans of 41 patients before ablation and at 24 hours and 3 months after ablation to determine whether there was evidence of contrast enhancement in the esophagus after atrial fibrillation ablation. In patients with contrast enhancement, 3D segmentation of the esophagus was performed using a novel image processing method. Upper gastrointestinal endoscopy was then performed. Repeat delayed-enhancement MRI and upper gastrointestinal endoscopy was performed 1 week later to track changes in lesions. The wall thickness of the anterior and posterior wall of the esophagus was measured at 3 time points: before ablation, 24 hours after ablation, and 3 months after ablation. Evaluation of preablation MRI scans demonstrated no cases of esophageal enhancement. At 24 hours, 5 patients showed contrast enhancement. Three of these patients underwent upper gastrointestinal endoscopy, which demonstrated esophageal lesions. Repeat upper gastrointestinal endoscopy and MRI 1 week later demonstrated resolution of the lesions. All 5 patients had confirmed resolution of enhancement at 3 months. All patients with esophageal tissue enhancement demonstrated left atrial wall enhancement directly adjacent to the regions of anterior wall esophageal enhancement. CONCLUSIONS: Our preliminary results indicate delayed-enhancement MRI can assess the extent and follow progression of esophageal wall injury after catheter ablation of atrial fibrillation. It appears that acute esophageal injury recovers within 1 week of the procedure. PMID- 20009077 TI - Pulmonary vein isolation for the maintenance of sinus rhythm in patients with atrial fibrillation: a meta-analysis of randomized, controlled trials. AB - BACKGROUND: Catheter ablation is an established yet evolving nonpharmacologic intervention for the maintenance of sinus rhythm in patients with atrial fibrillation (AF). The efficacy and safety of pulmonary vein isolation (PVI) compared with medical therapy remain in question. METHODS AND RESULTS: We conducted a meta-analysis of all randomized, controlled trials comparing PVI and medical therapy for the maintenance of sinus rhythm. The primary end point in this analysis was freedom from recurrent AF at 12 months. The relative efficacy of PVI was estimated using random-effects modeling according to intention to treat. We identified 6 trials that randomized a total of 693 patients with AF to PVI or control. PVI was associated with markedly increased odds of freedom from AF at 12 months of follow-up (n=266/344 [77%] versus n=102/346 [29%]; odds ratio, 9.74; 95% CI, 3.98 to 23.87). When we excluded the trial that only enrolled patients with persistent AF (Q-statistic, 2.485; P=0.647 after exclusion), PVI was associated with even greater odds of AF-free survival (15.78; 95% CI, 10.07 to 24.73). PVI was associated with a decreased hospitalization for cardiovascular causes (14 versus 93 per 100 person-years; rate ratio, 0.15; 95% CI, 0.10 to 0.23). Among those randomly assigned to PVI, 17% required a repeat PVI ablation before 12 months. The rate of major complications was 2.6% (n=9/344) in the catheter ablation group. CONCLUSIONS: Compared with a nonablation treatment strategy, PVI results in dramatically increased freedom from AF at 1 year. Although the procedure can be associated with major complications, the risk of these complications is comparable to other interventional procedures. PMID- 20009078 TI - Ventricular tachycardia arising from the aortomitral continuity in structural heart disease: characteristics and therapeutic considerations for an anatomically challenging area of origin. AB - BACKGROUND: The aortomitral continuity (AMC) has been described as a site of origin for ventricular tachycardias (VT) in structurally normal hearts. There is a paucity of data on the contribution of this region to VTs in patients with structural heart disease. METHODS AND RESULTS: Data from 550 consecutive patients undergoing catheter ablation for VT associated with structural heart disease were reviewed. Twenty-one (3.8%) had a VT involving the peri-AMC region (age, 62.7+/ 11 years; median left ventricular ejection fraction, 43.6+/-17%). Structural heart disease was ischemic in 7 (33%), dilated cardiomyopathy in 10 (47.6%), and valvular cardiomyopathy in 4 (19%) patients, respectively. After 1.9+/-0.8 catheter ablation procedures (including 3 transcoronary ethanol ablations) the peri-AMC VT was not inducible in 19 patients. The remaining 2 patients underwent cryosurgical ablation. Our first catheter ablation procedure was less often successful (66.7%) for peri-AMC VTs compared with that for 246 VTs originating from the LV free wall (81.4%, P=0.03). During a mean follow-up of 1.9+/-2.1 years, 12 (57.1%) patients remained free of VT, peri-AMC VT recurred in 7 patients, and 1 patient had recurrent VT from a remote location. Three patients died. Analysis of 50 normal coronary angiograms demonstrated an early septal branch supplying the peri-AMC area in 58% of cases that is a potential target for ethanol ablation. CONCLUSIONS: VTs involving the peri-AMC region occur in patients with structural heart disease and appear to be more difficult to ablate compared with VTs originating from the free LV wall. This region provides unique challenges for radiofrequency ablation, but cryosurgery and transcoronary alcohol ablation appear feasible in some cases. PMID- 20009079 TI - Alpha1-syntrophin mutations identified in sudden infant death syndrome cause an increase in late cardiac sodium current. AB - BACKGROUND: Sudden infant death syndrome (SIDS) is a leading cause of death during the first 6 months after birth. About 5% to 10% of SIDS may stem from cardiac channelopathies such as long-QT syndrome. We recently implicated mutations in alpha1-syntrophin (SNTA1) as a novel cause of long-QT syndrome, whereby mutant SNTA1 released inhibition of associated neuronal nitric oxide synthase by the plasma membrane Ca-ATPase PMCA4b, causing increased peak and late sodium current (I(Na)) via S-nitrosylation of the cardiac sodium channel. This study determined the prevalence and functional properties of SIDS-associated SNTA1 mutations. METHODS AND RESULTS: Using polymerase chain reaction, denaturing high-performance liquid chromatography, and DNA sequencing of SNTA1's open reading frame, 6 rare (absent in 800 reference alleles) missense mutations (G54R, P56S, T262P, S287R, T372M, and G460S) were identified in 8 (approximately 3%) of 292 SIDS cases. These mutations were engineered using polymerase chain reaction based overlap extension and were coexpressed heterologously with SCN5A, neuronal nitric oxide synthase, and PMCA4b in HEK293 cells. I(Na) was recorded using the whole-cell method. A significant 1.4- to 1.5-fold increase in peak I(Na) and 2.3- to 2.7-fold increase in late I(Na) compared with controls was evident for S287R-, T372M-, and G460S-SNTA1 and was reversed by a neuronal nitric oxide synthase inhibitor. These 3 mutations also caused a significant depolarizing shift in channel inactivation, thereby increasing the overlap of the activation and inactivation curves to increase window current. CONCLUSIONS: Abnormal biophysical phenotypes implicate mutations in SNTA1 as a novel pathogenic mechanism for the subset of channelopathic SIDS. Functional studies are essential to distinguish pathogenic perturbations in channel interacting proteins such as alpha1 syntrophin from similarly rare but innocuous ones. PMID- 20009081 TI - Should catheter ablation be the preferred therapy for reducing ICD shocks?: Ventricular tachycardia ablation versus drugs for preventing ICD shocks: role of adjuvant antiarrhythmic drug therapy. PMID- 20009080 TI - Sudden infant death syndrome in mice with an inherited mutation in RyR2. AB - BACKGROUND: Mutations in the cardiac ryanodine receptor gene (RyR2) have been recently identified in victims of sudden infant death syndrome. The aim of this study was to determine whether a gain-of-function mutation in RyR2 increases the propensity to cardiac arrhythmias and sudden death in young mice. METHODS AND RESULTS: Incidence of sudden death was monitored prospectively in heterozygous knock-in mice with mutation R176Q in RyR2 (R176Q/+). Young R176Q/+ mice exhibited a higher incidence of sudden death compared with wild-type littermates. Optical mapping of membrane potentials and intracellular calcium in 1- to 7-day-old R176Q/+ and wild-type mice revealed an increased incidence of ventricular ectopy and spontaneous calcium releases in neonatal R176Q/+ mice. Surface ECGs in 3- to 10-day-old mice showed that R176Q/+ mice developed more ventricular arrhythmias after provocation with epinephrine and caffeine. Intracardiac pacing studies in 12- to 18-day-old mice revealed the presence of an arrhythmogenic substrate in R176Q/+ compared with wild-type mice. Reverse transcription-polymerase chain reaction and Western blotting showed that expression levels of other calcium handling proteins were unaltered, suggesting that calcium leak through mutant RyR2 underlies arrhythmogenesis and sudden death in young R176Q/+ mice. CONCLUSIONS: Our findings demonstrate that a gain-of-function mutation in RyR2 confers an increased risk of cardiac arrhythmias and sudden death in young mice and that young R176Q/+ mice may be used as a model for elucidating the complex interplay between genetic and environmental risk factors associated with sudden infant death syndrome. PMID- 20009082 TI - Should catheter ablation be the preferred therapy for reducing ICD shocks?: Ventricular tachycardia in patients with an implantable defibrillator warrants catheter ablation. PMID- 20009083 TI - Plasma membrane expression of gonadotropin-releasing hormone receptors: regulation by peptide and nonpeptide antagonists. AB - Gonadotropin-releasing hormone acts via cell surface receptors but most human (h) GnRH receptors (GnRHRs) are intracellular. A membrane-permeant nonpeptide antagonist [(2S)-2-[5-[2-(2-axabicyclo[2.2.2]oct-2-yl)-1,1-dimethy-2-oxoethyl]-2 (3,5-dimethylphenyl)-1H-indol-3-yl]-N-(2-pyridin-4-ylethyl)propan-1-amine (IN3)] increases hGnRHR expression at the surface, apparently by facilitating its exit from the endoplasmic reticulum. Here we have quantified GnRHR by automated imaging in HeLa cells transduced with adenovirus expressing hemagglutinin-tagged GnRHR. Consistent with an intracellular site of action, IN3 increases cell surface hGnRHR, and this effect is not blocked or mimicked by membrane-impermeant peptide antagonists [Ac-D2Nal-D4Cpa-D3Pal-Ser-Tyr-d-Cit-Leu-Arg-Pro-d-Ala-NH(2) (cetrorelix) and antide]. However, when the C-terminal tail of a Xenopus (X) GnRHR was added (h.XGnRHR) to increase expression, both peptides further increased cell surface GnRHR. Cetrorelix also synergized with IN3 to increase expression of hGnRHR and a G-protein coupling-deficient mutant (A261K-hGnRHR). Cetrorelix also increased cell surface expression of hGnRHR, h.XGnRHR, and mouse GnRHR in gonadotrope-lineage LbetaT2 cells, and in HeLa cells it slowed h.XGnRHR internalization (measured by receptor-mediated antihemagglutinin uptake). Thus cetrorelix has effects other than GnRHR blockade; it acts as an inverse agonist in internalization assays, supporting the potential importance of ligand-biased efficacy at GnRHR. We also developed an imaging assay for GnRH function based on Ca(2+)-dependent nuclear translocation of a nuclear factor of activated T cells reporter. Using this in HeLa and LbetaT2 cells, IN3 and cetrorelix behaved as competitive antagonists when coincubated with GnRH, and long-term pretreatment (16 h) with IN3 reduced its effectiveness as an inhibitor whereas pretreatment with cetrorelix increased its inhibitory effect. This distinction between peptide and nonpeptide antagonists may prove important for therapeutic applications of GnRH antagonists. PMID- 20009084 TI - Sustained activity of the EGF receptor is an absolute requisite for LH-induced oocyte maturation and cumulus expansion. AB - Mammalian reproduction depends on the release of a mature oocyte from the ovarian follicle. Maturation of the oocyte and rupture of the follicle wall constitute part of the responses to the preovulatory surge of LH, which also include cumulus expansion and granulosa cell luteinization. It was previously shown that the epidermal growth factor receptor (EGFR) mediates the ovulatory response to LH in the ovarian follicle. We hypothesized that it is a sustained activity of the EGFR that generates oocyte maturation and cumulus expansion. We demonstrated that, whereas a transient exposure of rat isolated, intact, preovulatory follicles to either LH or forskolin was sufficient to induce oocyte maturation and cumulus expansion, these LH-induced responses were only generated upon a prolonged activity of the EGFR. In addition, the continuous activity of the EGFR is essential for the chronic phosphorylation of the ERK1/2 downstream signaling molecules, which were shown to be essential for oocyte maturation and cumulus expansion. Interestingly, EGFR-sustained activity was also necessary to maintain the up-regulation of Ptgs2, a gene essential for cumulus expansion. The unusual prolonged duration of ERK1/2 activity may possibly be attributed to the late induction of the ERK-specific phosphatase 3, demonstrated herein. These new data shed light on the unique characteristics of EGFR-ERK1/2 activity in the ovarian follicle and emphasize the fact that the ovulatory process involves a nonclassical activation of this pathway. PMID- 20009085 TI - Increased vitreous shedding of microparticles in proliferative diabetic retinopathy stimulates endothelial proliferation. AB - OBJECTIVE: Diabetic retinopathy is associated with progressive retinal capillary activation and proliferation, leading to vision impairment and blindness. Microparticles are submicron membrane vesicles with biological activities, released following cell activation or apoptosis. We tested the hypothesis that proangiogenic microparticles accumulate in vitreous fluid in diabetic retinopathy. RESEARCH DESIGN AND METHODS: Levels and cellular origin of vitreous and plasma microparticles from control (n = 26) and diabetic (n = 104) patients were analyzed by flow cytometry, and their proangiogenic activity was assessed by in vitro thymidine incorporation and neovessel formation in subcutaneous Matrigel plugs in mice. RESULTS: Microparticles of endothelial, platelet, photoreceptor, and microglial origin were identified in vitreous samples. Levels of photoreceptor and microglial microparticles were undetectable in plasmas but were comparable in diabetic and control vitreous samples. Vitreous platelet and endothelial microparticles levels were increased in diabetic patients and decreased following panretinal laser photocoagulation or intravitreal antivascular endothelial growth factor injection in proliferative diabetic retinopathy (PDR). The ratio of vitreous to plasma microparticle levels was calculated to estimate local formation versus potential plasma leakage. In PDR, the endothelial microparticles ratio--but not that for platelet--was greater than 1.0, indicating local formation of endothelial microparticles from retinal vessels and permeation of platelet microparticles from plasma. Isolated vitreous microparticles stimulated by 1.6-fold endothelial proliferation and increased new vessel formation in mice. CONCLUSIONS: The present study demonstrates that vitreous fluid contains shed membrane microparticles of endothelial, platelet, and retinal origin. Vitreous microparticles levels are increased in patients with diabetic retinopathy, where they could contribute to disease progression. PMID- 20009086 TI - A novel hypomorphic PDX1 mutation responsible for permanent neonatal diabetes with subclinical exocrine deficiency. AB - OBJECTIVE: Genes responsible for monogenic forms of diabetes have proven very valuable for understanding key mechanisms involved in beta-cell development and function. Genetic study of selected families is a powerful strategy to identify such genes. We studied a consanguineous family with two first cousins affected by neonatal diabetes; their four parents had a common ancestor, suggestive of a fully penetrant recessive mutation. RESEARCH DESIGN AND METHODS: We performed genetic studies of the family, detailed clinical and biochemical investigations of the patients and the four parents, and biochemical and functional studies of the new mutation. RESULTS: We found a novel mutation in the pancreatic and duodenal homeobox 1 gene (PDX1, IPF1) in the two patients, which segregated with diabetes in the homozygous state. The mutation resulted in an E178G substitution in the PDX1 homeodomain. In contrast to other reported PDX1 mutations leading to neonatal diabetes and pancreas agenesis, homozygosity for the E178G mutation was not associated with clinical signs of exocrine pancreas insufficiency. Further, the four heterozygous parents were not diabetic and displayed normal glucose tolerance. Biochemical studies, however, revealed subclinical exocrine pancreas insufficiency in the patients and slightly reduced insulin secretion in the heterozygous parents. The E178G mutation resulted in reduced Pdx1 transactivation despite normal nuclear localization, expression level, and chromatin occupancy. CONCLUSIONS: This study broadens the clinical spectrum of PDX1 mutations and justifies screening of this gene in neonatal diabetic patients even in the absence of exocrine pancreas manifestations. PMID- 20009087 TI - Genetic variation at the FTO locus influences RBL2 gene expression. AB - OBJECTIVE: Genome-wide association studies that compare the statistical association between thousands of DNA variations and a human trait have detected 958 loci across 127 different diseases and traits. However, these statistical associations only provide evidence for genomic regions likely to harbor a causal gene(s) and do not directly identify such genes. We combined gene variation and expression data in a human cohort to identify causal genes. RESEARCH DESIGN AND METHODS: Global gene transcription activity was obtained for each individual in a large human cohort (n = 1,240). These quantitative transcript data were tested for correlation with genotype data generated from the same individuals to identify gene expression patterns influenced by the variants. RESULTS: Variant rs8050136 lies within intron 1 of the FTO gene on chromosome 16 and marks a locus strongly associated with type 2 diabetes and obesity and widely replicated across many populations. We report that genetic variation at this locus does not influence FTO gene expression levels (P = 0.38), but is strongly correlated with expression of RBL2 (P = 2.7 x 10(-5)), approximately 270,000 base pairs distant to FTO. CONCLUSIONS: These data suggest that variants at FTO influence RBL2 gene expression at large genetic distances. This observation underscores the complexity of human transcriptional regulation and highlights the utility of large human cohorts in which both genetic variation and global gene expression data are available to identify disease genes. Expedient identification of genes mediating the effects of genome-wide association study-identified loci will enable mechanism-of-action studies and accelerate understanding of human disease processes under genetic influence. PMID- 20009088 TI - Hyperglycemia impairs proteasome function by methylglyoxal. AB - OBJECTIVE: The ubiquitin-proteasome system is the main degradation machinery for intracellularly altered proteins. Hyperglycemia has been shown to increase intracellular levels of the reactive dicarbonyl methylglyoxal (MGO) in cells damaged by diabetes, resulting in modification of proteins and alterations of their function. In this study, the influence of MGO-derived advanced glycation end product (AGE) formation on the activity of the proteasome was investigated in vitro and in vivo. RESEARCH DESIGN AND METHODS: MGO-derived AGE modification of proteasome subunits was analyzed by mass spectrometry, immunoprecipitation, and Western blots. Proteasome activity was analyzed using proteasome-specific fluorogenic substrates. Experimental models included bovine retinal endothelial cells, diabetic Ins2(Akita) mice, glyoxalase 1 (GLO1) knockdown mice, and streptozotocin (STZ)-injected diabetic mice. RESULTS: In vitro incubation with MGO caused adduct formation on several 20S proteasomal subunit proteins. In cultured endothelial cells, the expression level of the catalytic 20S proteasome subunit was not altered but proteasomal chymotrypsin-like activity was significantly reduced. In contrast, levels of regulatory 19S proteasomal proteins were decreased. In diabetic Ins2(Akita), STZ diabetic, and nondiabetic and diabetic G101 knockdown mice, chymotrypsin-like activity was also reduced and MGO modification of the 20S-beta2 subunit was increased. CONCLUSIONS: Hyperglycemia induced formation of MGO covalently modifies the 20S proteasome, decreasing its activity in the diabetic kidney and reducing the polyubiquitin receptor 19S-S5a. The results indicate a new link between hyperglycemia and impairment of cell functions. PMID- 20009089 TI - Oxygen tension regulates pancreatic beta-cell differentiation through hypoxia inducible factor 1alpha. AB - OBJECTIVE: Recent evidence indicates that low oxygen tension (pO2) or hypoxia controls the differentiation of several cell types during development. Variations of pO2 are mediated through the hypoxia-inducible factor (HIF), a crucial mediator of the adaptative response of cells to hypoxia. The aim of this study was to investigate the role of pO2 in beta-cell differentiation. RESEARCH DESIGN AND METHODS: We analyzed the capacity of beta-cell differentiation in the rat embryonic pancreas using two in vitro assays. Pancreata were cultured either in collagen or on a filter at the air/liquid interface with various pO2. An inhibitor of the prolyl hydroxylases, dimethyloxaloylglycine (DMOG), was used to stabilize HIF1alpha protein in normoxia. RESULTS: When cultured in collagen, embryonic pancreatic cells were hypoxic and expressed HIF1alpha and rare beta cells differentiated. In pancreata cultured on filter (normoxia), HIF1alpha expression decreased and numerous beta-cells developed. During pancreas development, HIF1alpha levels were elevated at early stages and decreased with time. To determine the effect of pO2 on beta-cell differentiation, pancreata were cultured in collagen at increasing concentrations of O2. Such conditions repressed HIF1alpha expression, fostered development of Ngn3-positive endocrine progenitors, and induced beta-cell differentiation by O2 in a dose-dependent manner. By contrast, forced expression of HIF1alpha in normoxia using DMOG repressed Ngn3 expression and blocked beta-cell development. Finally, hypoxia requires hairy and enhancer of split (HES)1 expression to repress beta-cell differentiation. CONCLUSIONS: These data demonstrate that beta-cell differentiation is controlled by pO2 through HIF1alpha. Modifying pO2 should now be tested in protocols aiming to differentiate beta-cells from embryonic stem cells. PMID- 20009090 TI - Effect of rimonabant on glycemic control in insulin-treated type 2 diabetes: the ARPEGGIO trial. AB - OBJECTIVE To examine the efficacy and safety of rimonabant, a selective cannabinoid receptor type-1 antagonist, in patients with type 2 diabetes receiving insulin monotherapy. RESEARCH DESIGN AND METHODS Patients (n = 368; A1C > or =7%) were randomized to 20 mg/day rimonabant or placebo in this 48-week, double-blind, placebo-controlled multicenter trial. Change in baseline A1C to week 48 (primary outcome) and changes in body weight, waist circumference, and lipid levels (secondary outcomes) were assessed. RESULTS Rimonabant significantly reduced baseline A1C versus placebo (-0.89 vs. -0.24%; P < 0.0001), and significantly greater improvements were observed in cardiometabolic risk factors. More rimonabant patients achieved >10% reduction in mean total daily insulin dose versus placebo (P = 0.0012), and fewer required rescue medication (P < 0.0001). Hypoglycemia, nausea, dizziness, anxiety, and depression were more frequent with rimonabant. CONCLUSIONS Rimonabant improved glycemic control and cardiometabolic risk factors in patients with type 2 diabetes receiving insulin. PMID- 20009091 TI - Active care management supported by home telemonitoring in veterans with type 2 diabetes: the DiaTel randomized controlled trial. AB - OBJECTIVE We compared the short-term efficacy of home telemonitoring coupled with active medication management by a nurse practitioner with a monthly care coordination telephone call on glycemic control in veterans with type 2 diabetes and entry A1C > or =7.5%. RESEARCH DESIGN AND METHODS Veterans who received primary care at the VA Pittsburgh Healthcare System from June 2004 to December 2005, who were taking oral hypoglycemic agents and/or insulin for > or =1 year, and who had A1C > or =7.5% at enrollment were randomly assigned to either active care management with home telemonitoring (ACM+HT group, n = 73) or a monthly care coordination telephone call (CC group, n = 77). Both groups received monthly calls for diabetes education and self-management review. ACM+HT group participants transmitted blood glucose, blood pressure, and weight to a nurse practitioner using the Viterion 100 TeleHealth Monitor; the nurse practitioner adjusted medications for glucose, blood pressure, and lipid control based on established American Diabetes Association targets. Measures were obtained at baseline, 3-month, and 6-month visits. RESULTS Baseline characteristics were similar in both groups, with mean A1C of 9.4% (CC group) and 9.6% (ACM+HT group). Compared with the CC group, the ACM+HT group demonstrated significantly larger decreases in A1C at 3 months (1.7 vs. 0.7%) and 6 months (1.7 vs. 0.8%; P < 0.001 for each), with most improvement occurring by 3 months. CONCLUSIONS Compared with the CC group, the ACM+HT group demonstrated significantly greater reductions in A1C by 3 and 6 months. However, both interventions improved glycemic control in primary care patients with previously inadequate control. PMID- 20009092 TI - Thoracoabdominal calcifications predict cardiovascular disease mortality in type 2 diabetic and nondiabetic subjects: 18-year follow-up study. AB - OBJECTIVE To evaluate cardiovascular disease (CVD) and total mortality associated with thoracoabdominal calcifications. RESEARCH DESIGN AND METHODS Thoracoabdominal calcifications of native radiograms were evaluated in 833 subjects with type 2 diabetes and 1,292 subjects without diabetes, aged 45-64 years, without prior evidence of CVD. The type 2 diabetic and nondiabetic study cohorts were followed up for 18 years. RESULTS After adjustment for conventional risk factors, marked thoracoabdominal calcifications predicted CVD/total mortality with hazard ratio (HR) (95% CI) of 1.5 (0.8-3.0)/1.8 (1.1-2.9) in type 2 diabetic men, 3.0 (1.6-5.7)/3.1 (1.9-5.0) in type 2 diabetic women, 5.0 (2.2 12)/4.0 (2.2-7.4) in nondiabetic men, and 7.8 (1.8-34)/3.0 (1.3-7.0) in nondiabetic women and in the presence of C-reactive protein below/over 3 mg/l with HR of 2.4 (1.3-4.4)/3.0 (1.4-6.1) in type 2 diabetic subjects and 4.0 (1.5 10.8)/6.6 (2.7-16.0) in nondiabetic subjects. CONCLUSIONS Thoracoabdominal calcifications in native radiograms are significant predictors of CVD and total mortality, especially in type 2 diabetic and nondiabetic women with elevated high sensitivity C-reactive protein level. PMID- 20009093 TI - Rapid identification of myocardial infarction risk associated with diabetes medications using electronic medical records. AB - OBJECTIVE To assess the ability to identify potential association(s) of diabetes medications with myocardial infarction using usual care clinical data obtained from the electronic medical record. RESEARCH DESIGN AND METHODS We defined a retrospective cohort of patients (n = 34,253) treated with a sulfonylurea, metformin, rosiglitazone, or pioglitazone in a single academic health care network. All patients were aged >18 years with at least one prescription for one of the medications between 1 January 2000 and 31 December 2006. The study outcome was acute myocardial infarction requiring hospitalization. We used a cumulative temporal approach to ascertain the calendar date for earliest identifiable risk associated with rosiglitazone compared with that for other therapies. RESULTS Sulfonylurea, metformin, rosiglitazone, or pioglitazone therapy was prescribed for 11,200, 12,490, 1,879, and 806 patients, respectively. A total of 1,343 myocardial infarctions were identified. After adjustment for potential myocardial infarction risk factors, the relative risk for myocardial infarction with rosiglitazone was 1.3 (95% CI 1.1-1.6) compared with sulfonylurea, 2.2 (1.6-3.1) compared with metformin, and 2.2 (1.5-3.4) compared with pioglitazone. Prospective surveillance using these data would have identified increased risk for myocardial infarction with rosiglitazone compared with metformin within 18 months of its introduction with a risk ratio of 2.1 (95% CI 1.2-3.8). CONCLUSIONS Our results are consistent with a relative adverse cardiovascular risk profile for rosiglitazone. Our use of usual care electronic data sources from a large hospital network represents an innovative approach to rapid safety signal detection that may enable more effective postmarketing drug surveillance. PMID- 20009094 TI - Patient race/ethnicity and patient-physician race/ethnicity concordance in the management of cardiovascular disease risk factors for patients with diabetes. AB - OBJECTIVE Patient-physician race/ethnicity concordance can improve care for minority patients. However, its effect on cardiovascular disease (CVD) care and prevention is unknown. We examined associations of patient race/ethnicity and patient-physician race/ethnicity concordance on CVD risk factor levels and appropriate modification of treatment in response to high risk factor values (treatment intensification) in a large cohort of diabetic patients. RESEARCH DESIGN AND METHODS The study population included 108,555 adult diabetic patients in Kaiser Permanente Northern California in 2005. Probit models assessed the effect of patient race/ethnicity on risk factor control and treatment intensification after adjusting for patient and physician-level characteristics. RESULTS African American patients were less likely than whites to have A1C <8.0% (64 vs. 69%, P < 0.0001), LDL cholesterol <100 mg/dl (40 vs. 47%, P < 0.0001), and systolic blood pressure (SBP) <140 mmHg (70 vs. 78%, P < 0.0001). Hispanic patients were less likely than whites to have A1C <8% (62 vs. 69%, P < 0.0001). African American patients were less likely than whites to have A1C treatment intensification (73 vs. 77%, P < 0.0001; odds ratio [OR] 0.8 [95% CI 0.7-0.9]) but more likely to receive treatment intensification for SBP (78 vs. 71%, P < 0.0001; 1.5 [1.3-1.7]). Hispanic patients were more likely to have LDL cholesterol treatment intensification (47 vs. 45%, P < 0.05; 1.1 [1.0-1.2]). Patient-physician race/ethnicity concordance was not significantly associated with risk factor control or treatment intensification. CONCLUSIONS Patient race/ethnicity is associated with risk factor control and treatment intensification, but patient-physician race/ethnicity concordance was not. Further research should investigate other potential drivers of disparities in CVD care. PMID- 20009095 TI - Effect of Rosiglitazone and Ramipril on {beta}-cell function in people with impaired glucose tolerance or impaired fasting glucose: the DREAM trial. AB - OBJECTIVE The objective of this study was to determine the degree to which ramipril and/or rosiglitazone changed beta-cell function over time among individuals with impaired fasting glucose (IFG) and/or impaired glucose tolerance (IGT) who participated in the Diabetes Reduction Assessment With Ramipril and Rosiglitazone Medication (DREAM) Trial, which evaluated whether ramipril and/or rosiglitazone could prevent or delay type 2 diabetes in high-risk individuals. RESEARCH DESIGN AND METHODS The present analysis included subjects (n = 982) from DREAM trial centers in Canada who had oral glucose tolerance tests at baseline, after 2 years, and at the end of the study. beta-Cell function was assessed using the fasting proinsulin-to-C-peptide ratio (PI/C) and the insulinogenic index (defined as 30-0 min insulin/30-0 min glucose) divided by homeostasis model assessment of insulin resistance (insulinogenic index [IGI]/insulin resistance [IR]). RESULTS Subjects receiving rosiglitazone had a significant increase in IGI/IR between baseline and end of study compared with the placebo group (25.59 vs. 1.94, P < 0.0001) and a significant decrease in PI/C (-0.010 vs. -0.006, P < 0.0001). In contrast, there were no significant changes in IGI/IR or PI/C in subjects receiving ramipril compared with placebo (11.71 vs. 18.15, P = 0.89, and -0.007 vs. -0.008, P = 0.64, respectively). The impact of rosiglitazone on IGI/IR and PI/C was similar within subgroups of isolated IGT and IFG + IGT (all P < 0.001). Effects were more modest in those with isolated IFG (IGI/IR: 8.95 vs. 2.13, P = 0.03; PI/C: -0.003 vs. -0.001, P = 0.07). CONCLUSIONS Treatment with rosiglitazone, but not ramipril, resulted in significant improvements in measures of beta-cell function over time in pre-diabetic subjects. Although the long-term sustainability of these improvements cannot be determined from the present study, these findings demonstrate that the diabetes preventive effect of rosiglitazone was in part a consequence of improved beta-cell function. PMID- 20009096 TI - Utility of childhood glucose homeostasis variables in predicting adult diabetes and related cardiometabolic risk factors: the Bogalusa Heart Study. AB - OBJECTIVE This study examines the usefulness of childhood glucose homeostasis variables (glucose, insulin, and insulin resistance index [homeostasis model assessment of insulin resistance {HOMA-IR}]) in predicting pre-diabetes and type 2 diabetes and related cardiometabolic risk factors in adulthood. RESEARCH DESIGN AND METHODS This retrospective cohort study consisted of normoglycemic (n = 1,058), pre-diabetic (n = 37), and type 2 diabetic (n = 25) adults aged 19-39 years who were followed on average for 17 years since childhood. RESULTS At least 50% of the individuals who ranked highest (top quintile) in childhood for glucose homeostasis variables maintained their high rank by being above the 60th percentile in adulthood. In a multivariate model, the best predictors of adulthood glucose homeostasis variables were the change in BMI Z score from childhood to adulthood and childhood BMI Z score, followed by the corresponding childhood levels of glucose, insulin, and HOMA-IR. Further, children in the top decile versus the rest for insulin and HOMA-IR were 2.85 and 2.55 times, respectively, more likely to develop pre-diabetes; children in the top decile versus the rest for glucose, insulin, and HOMA-IR were 3.28, 5.54, and 5.84 times, respectively, more likely to develop diabetes, independent of change in BMI Z score, baseline BMI Z score, and total-to-HDL cholesterol ratio. In addition, children with adverse levels (top quintile versus the rest) of glucose homeostasis variables displayed significantly higher prevalences of, among others, hyperglycemia, hypertriglyceridemia, and metabolic syndrome. CONCLUSIONS Adverse levels of glucose homeostasis variables in childhood not only persist into adulthood but also predict adult pre-diabetes and type 2 diabetes and relate to cardiometabolic risk factors. PMID- 20009097 TI - Lipotoxicity and decreased islet graft survival. AB - OBJECTIVE To evaluate if baseline serum lipids are associated with islet graft survival in type 1 diabetes islet transplant (ITx) recipients. RESEARCH DESIGN AND METHODS Baseline fasting lipid profile was collected from 44 ITx recipients. Comparisons were performed between subjects below and above the median values of each lipid fraction. Differences in outcomes were compared by Kaplan-Meier curves and Cox regression analysis. RESULTS Subjects with baseline fasting plasma triglycerides and VLDL cholesterol above the median had shorter islet graft survival (triglycerides: 39.7 +/- 6.1 vs. 61.3 +/- 6.6 months, P = 0.029, and VLDL: 41.5 +/- 5.7 vs. 62.8 +/- 7.3 months, P = 0.032). Total, LDL, and HDL cholesterol did not influence islet function. Triglycerides (odds ratio 2.97 [95% CI 1.03-8.52], P = 0.044) maintained its association with graft failure after adjustments for confounders. CONCLUSIONS Higher baseline triglycerides are associated with earlier decline in islet graft function. Prospective clinical trials should address whether it is directly caused by lipotoxicity and if strategies focusing on lowering serum lipids may prolong islet graft survival. PMID- 20009098 TI - Carboxylation of osteocalcin affects its association with metabolic parameters in healthy children. AB - OBJECTIVE Osteocalcin (OC), a bone-derived protein, was recently shown to regulate metabolic pathways in mice. Undercarboxylated OC (ucOC), but not carboxylated OC (cOC), increases adiponectin and insulin secretion. It is unclear if carboxylation of OC affects its association with metabolic parameters in humans. RESEARCH DESIGN AND METHODS The associations between ucOC, cOC, total and high-molecular-weight (HMW) adiponectin, and insulin secretion (homeostasis model assessment [HOMA]-beta) were investigated in a population-based sample of healthy prepubertal children (n = 103; 49 boys and 54 girls). RESULTS Weight-dependent associations were observed between the different forms of OC and metabolic parameters. Higher cOC was related to lower HMW adiponectin (with a stronger association in leaner children; P < 0.001). Higher ucOC-to-cOC ratio was associated with higher HOMA-beta (P < 0.01) in leaner children and associated with higher HMW adiponectin (P < 0.001) in heavier children. CONCLUSIONS In a weight-dependent manner, cOC and the proportion of ucOC are differentially related to HMW adiponectin and insulin secretion in healthy children. PMID- 20009099 TI - Moving to an A1C-based diagnosis of diabetes has a different impact on prevalence in different ethnic groups. AB - OBJECTIVE To compare screen-detected diabetes prevalence and the degree of diagnostic agreement by ethnicity with the current oral glucose tolerance test (OGTT)-based and newly proposed A1C-based diagnostic criteria. RESEARCH DESIGN AND METHODS Six studies (1999-2009) from Denmark, the U.K., Australia, Greenland, Kenya, and India were tested for the probability of an A1C > or =6.5% among diabetic case subjects based on an OGTT. The difference in probability between centers was analyzed by logistic regression adjusting for relevant confounders. RESULTS Diabetes prevalence was lower with the A1C-based diagnostic criteria in four of six studies. The probability of an A1C > or =6.5% among OGTT-diagnosed case subjects ranged widely (17.0-78.0%) by study center. Differences in diagnostic agreement between ethnic subgroups in the U.K. study were of the same magnitude as between-country comparisons. CONCLUSIONS A shift to an A1C-based diagnosis for diabetes will have substantially different consequences for diabetes prevalence across ethnic groups and populations. PMID- 20009100 TI - Mortality trends in subjects with and without diabetes during 33 years of follow up. AB - OBJECTIVE Mortality rates have declined substantially over the past decades in the general population, but the situation among diabetic subjects is less clear. The aim of this study was to analyze mortality trends in diabetic and nondiabetic subjects during 1972-2004. RESEARCH DESIGN AND METHODS Since 1972, all patients with diabetes are entered in a diabetes register at Laxa Primary Health Care Center; 776 incident cases were recorded up to 2001. The register has been supplemented with a nondiabetic population of 3,880 subjects and with data from the National Cause of Death Register during 1972 to 2004. RESULTS During the 33 year follow-up period, 233 (62.0%) diabetic women and 240 (60.0%) diabetic men and 995 (52.9%) nondiabetic women and 1,082 (54.1%) nondiabetic men died. The age adjusted hazard ratio (HR) for all-cause mortality among diabetic and nondiabetic subjects was 1.17 (P < 0.0021) for all, 1.22 (P < 0.007) for women, and 1.13 (P = 0.095) for men. The corresponding cardiovascular disease (CVD) mortality HRs were 1.33 (P < 0.0001), 1.41 (P < 0.0003), and 1.27 (P < 0.0093), respectively. The CVD mortality reduction across time was significant in nondiabetic subjects (P < 0.0001) and in men with diabetes (P = 0.014) but not in diabetic women (P = 0.69). The results regarding coronary heart disease (CHD) were similar (P < 0.0001, P < 0.006, and P = 0.17, respectively). The CVD and CHD mortality rate change across time was fairly linear in all groups. CONCLUSIONS Diabetic subjects had less mortality rate reduction during follow-up than nondiabetic subjects. However the excess mortality risk for diabetic subjects was smaller than that found in other studies. PMID- 20009102 TI - Cell mechanics and feedback regulation of actomyosin networks. AB - Actomyosin contractility is the major force-generating machinery that shapes cells and tissues during morphogenesis. New evidence from Drosophila demonstrates that these forces are spatially organized by a combination of biochemical and mechanical signals that provide dynamic feedback in a complex cellular environment. PMID- 20009104 TI - EGFR signaling through an Akt-SREBP-1-dependent, rapamycin-resistant pathway sensitizes glioblastomas to antilipogenic therapy. AB - Glioblastoma, the most common malignant brain tumor, is among the most lethal and difficult cancers to treat. Although epidermal growth factor receptor (EGFR) mutations are frequent in glioblastoma, their clinical relevance is poorly understood. Studies of tumors from patients treated with the EGFR inhibitor lapatinib revealed that EGFR induces the cleavage and nuclear translocation of the master transcriptional regulator of fatty acid synthesis, sterol regulatory element-binding protein 1 (SREBP-1). This response was mediated by Akt; however, clinical data from rapamycin-treated patients showed that SREBP-1 activation was independent of the mammalian target of rapamycin complex 1, possibly explaining rapamycin's poor efficacy in the treatment of such tumors. Glioblastomas without constitutively active EGFR signaling were resistant to inhibition of fatty acid synthesis, whereas introduction of a constitutively active mutant form of EGFR, EGFRvIII, sensitized tumor xenografts in mice to cell death, which was augmented by the hydroxymethylglutaryl coenzyme A reductase inhibitor atorvastatin. These results identify a previously undescribed EGFR-mediated prosurvival metabolic pathway and suggest new therapeutic approaches to treating EGFR-activated glioblastomas. PMID- 20009105 TI - Function of the nucleotide exchange activity of vav1 in T cell development and activation. AB - The guanine nucleotide exchange factor (GEF) Vav1 is essential for transducing T cell antigen receptor (TCR) signals and therefore plays a critical role in the development and activation of T cells. It has been presumed that the GEF activity of Vav1 is important for its function; however, there has been no direct demonstration of this. Here, we generated mice expressing enzymatically inactive, but normally folded, Vav1 protein. Analysis of these mice showed that the GEF activity of Vav1 was necessary for the selection of thymocytes and for the optimal activation of T cells, including signal transduction to Rac1, Akt, and integrins. In contrast, the GEF activity of Vav1 was not required for TCR-induced calcium flux, activation of extracellular signal-regulated kinase and protein kinase D1, and cell polarization. Thus, in T cells, the GEF activity of Vav1 is essential for some, but not all, of its functions. PMID- 20009111 TI - Personalised treatment of arthritis in the next eRA. PMID- 20009112 TI - Transcript profiling towards personalised medicine in rheumatoid arthritis. AB - Rheumatoid arthritis (RA ) is a chronic inflammatory joint disease that is heterogeneous in nature. The heterogeneity is reflected by the variation in responsiveness to virtually any treatment modality. Since our understanding of the molecular complexity is incomplete and criteria for categorisation are limited, we mainly consider the disease RA as group average. A powerful way to gain insight into the complexity of RA has arisen from DNA microarray technology, which allows an open-ended survey to comprehensively identify the genes and biological pathways that are associated with clinically defined conditions. During the last decade encouraging results have been generated towards the molecular description of complex diseases in general. Here, I describe developments in genomics research that provide a framework to increase our understanding of the pathogenesis and the identification of biomarkers for early diagnosis, prognosis and stratification, aimed at a personal medicine approach in RA . PMID- 20009113 TI - Noninvasive haemodynamic monitoring using finger arterial pressure waveforms. AB - Haemodynamic monitoring may potentially lead to improved quality of care in haemodynamic compromised patients. However, the usefulness of invasive techniques using the pulmonary artery catheter is questioned. Noninvasive techniques which provide data on haemodynamics might provide a good alternative. New techniques have been developed in recent years to monitor cardiac output and other parameters of cardiac performance continuously and noninvasively. Recently, a new technique has become available that assesses these haemodynamic data from finger arterial pressure waveforms obtained noninvasively. Although an invasively derived calibration is still needed to obtain absolute data on cardiac output, relative changes in cardiac output can be accurately monitored using this method. Currently, the device can be used in patients to continuously monitor haemodynamic data and guide therapy. Furthermore, it might have a role in clinical research to noninvasively assess cardiac output, as a surrogate endpoint, before and after interventions. Although this new method seems promising, the clinical value has to be proven. PMID- 20009114 TI - Type 1 diabetes and autoimmune polyglandular syndrome: a clinical review. AB - Type 1 diabetes mellitus (T1DM) results from autoimmune destruction of insulin producing beta cells and is characterised by the presence of insulitis and &and beta-cell autoantibodies. Up to one third of patients develop an autoimmune polyglandular syndrome. Fifteen to 30% of T1DM subjects have autoimmune thyroid disease (Hashimoto's or Graves' disease), 5 to 10% are diagnosed with autoimmune gastritis and/or pernicious anaemia (AIG /PA), 4 to 9% present with coeliac disease (CD), 0.5% have Addison's disease (AD), and 2 to 10% show vitiligo. These diseases are characterised by the presence of autoantibodies against thyroid peroxidase (for Hashimoto's thyroiditis), TSH receptor (for Graves' disease), parietal cell or intrinsic factor (for AIG /PA), tissue transglutaminase (for CD), and 21-hydroxylase (for AD). Early detection of antibodies and latent organ specific dysfunction is advocated to alert physicians to take appropriate action in order to prevent full-blown disease. Hashimoto's hypothyroidism may cause weight gain, hyperlipidaemia, goitre, and may affect diabetes control, menses, and pregnancy outcome. In contrast, Graves' hyperthyroidism may induce weight loss, atrial fibrillation, heat intolerance, and ophthalmopathy. Autoimmune gastritis may manifest via iron deficiency or vitamin B12 deficiency anaemia with fatigue and painful neuropathy. Clinical features of coeliac disease include abdominal discomfort, growth abnormalities, infertility, low bone mineralisation, and iron deficiency anaemia. Adrenal insufficiency may cause vomiting, anorexia, hypoglycaemia, malaise, fatigue, muscular weakness, hyperkalaemia, hypotension, and generalised hyperpigmentation. Here we will review prevalence, pathogenetic factors, clinical features, and suggestions for screening, follow-up and treatment of patients with T1DM and/or autoimmune polyglandular syndrome. PMID- 20009115 TI - Computer-aided support improves early and adequate delivery of nutrients in the ICU. AB - BACKGROUND: In 2007 a national guideline on perioperative nutrition was issued in the Netherlands. As external indicator for adequacy of nutritional therapy, the percentage of malnourished patients who reach at least 1.2 grams of protein on day 4 after admission was chosen by the Netherlands Health Care Inspectorate. METHODS: We developed an algorithm that allows users to ask for advice on which artificial nutritional formula to prescribe and at which rate, assuring provision of adequate amounts of both protein and energy. Feedback on nutritional therapy is given to the users on a daily basis, and to the management per quarter. Both the advice and the feedback have been integrated in our data management system. The advice module is also available on-line. RESULTS: In the baseline situation over the first four quarters (2006) an average of 30.2% of patients who had a full day 4 in our unit reached the protein indicator. In the last six quarters post-implementation, the average percentage reached was 56.5% with values consistently over 50%. Changes were statistically significant at third quarter of 2007 (p<0.05) and thereafter (p<0.001). Results for day 7 of admission were unaffected, which indicates that targets were reached earlier during hospital stay. CONCLUSION: Our study shows that integration of nutritional advice and automatically generated feedback to users in a data management system consistently improves delivery of (early) nutrition. PMID- 20009116 TI - Glycogenic hepatopathy: a rare cause of elevated serum transaminases in diabetes mellitus. AB - Glycogenic hepatopathy (GH) is a rare cause of serum transaminase elevations in type 1 diabetes mellitus (DM). We describe a 29-year-old woman with a history of poorly controlled type 1 DM who presented with hepatomegaly and severe transaminase flares. Liver histology confirmed GH, with glycogen accumulation due to severe fluctuations in both glucose and insulin. GH can be regarded as an adult variant of Mauriac's syndrome. Despite severe laboratory abnormalities, it does not cause liver cirrhosis. Treatment consists of improving glycaemic control. PMID- 20009117 TI - A 79-year-old woman with incoercible vomiting. PMID- 20009118 TI - A patient with an inguinal mass: a groin hernia? PMID- 20009119 TI - Emphysematous pyelonephritis in a renal transplant patient. PMID- 20009120 TI - [Mastocytosis--diagnostic criteria and treatment]. AB - Mastocytosis is a heterogeneous group of rare diseases characterized by the proliferation and accumulation of mast cells in one or more organs such as the skin, bone marrow, liver, spleen, and lymph nodes. According to the WHO classification, mastocytosis is divided into seven subvariants. The symptoms are associated with mediator release and impaired organ function due to infiltration by neoplastic mast cells. There is a higher risk of anaphylactic shock; therefore education of the patients is very important. Patients may be asymptomatic. Symptomatic treatment is used in cutaneous mastocytosis and in indolent systemic mastocytosis. More aggressive subvariants of mastocytosis are treated with chemotherapy, targeted therapy, and bone marrow transplantation. PMID- 20009121 TI - [The genetics of dementias, Part 3: a molecular basis for the multifactorial inheritance of sporadic Alzheimer's disease]. AB - The majority of Alzheimer's disease cases, i.e. more than 85% of the whole population of patients, can be referred to as the sporadic form of the disease, with a negative family history and complex inheritance. As the genetic background of sporadic Alzheimer's disease is still largely unknown., strategies based on individual genetic risk profiling for either early prediction of the disease or its therapy and prevention are not possible. The APOE*4 allele of the gene for apolipoprotein E is still the only completely confirmed risk factor. Screening for new genetic risk factors with the use of genetic association analysis has many methodological difficulties and limitations. New combined approaches including genomics, proteogenomics, pharmacogenomics, epigenomics, and bioinformatics have to been applied in the future search for a molecular basis of AD. Genetic defects do not fully explain the complexity of the etiopathogenesis of this disease. It is rather certain that environmental factors (through epigenetic modifications in the patient' sepigenome) also have impact on the initiation of neurodegeneration processes. The identification of new genetic and environmental risk factors would make it possible to understand epistatic processes, for example interactions between genes and between genes and environmental factors, responsible for the complex etiology and multifactorial inheritance of sporadic Alzheimer's disease. PMID- 20009122 TI - [Genetics of dementias, Part 4: a spectrum of mutations responsible for the familial autosomal dominant form of Alzheimer's disease]. AB - Fifty years ago it was demonstrated that some patients with Alzheimer's disease (AD) had an autosomal dominant Mendelian pattern of disease inheritance. Familial and early-onset cases (familial Alzheimer's disease, FAD) are rather rare and account for only a few percent of the total population of patients. Mutations of the genes for amyloid precursor protein (APP), presenilin 1 (PSEN1), and presenilin 2 (PSEN2) are responsible for development of the disease in 50 percent of patients with FAD. The identification of mutations in FAD genes leads to a better understand of the molecular basis of the cellular pathways leading to neurodegeneration. With the detection of genetic defects responsible for FAD, there is considerable interest in the application of this genetic information in medical practice through genetic testing and counseling for families with Alzheimer's disease. PMID- 20009123 TI - [Chemical substances as risk factors of nephropathy in diabetes mellitus]. AB - Although diabetes mellitus, a metabolic disease, does not fall into the group of diseases induced by toxic substances or environmental pollution, there is much evidence that some chemicals have considerable importance in its development. Exposure to substances with potential renal toxicity is especially dangerous for diabetics because it accelerates and intensifies diabetic nephropathy. This paper discusses the relationship between the xenobiotics and the development of diabetes mellitus and diabetic nephropathy with particular emphasis on those substances that causes the greatest damage to the kidneys. These are cadmium, iron, lead, arsenic, polychlorinated organic compounds, nitrogen compounds, and contrast agents. In addition, the mechanisms of diabetes mellitus induction or kidney damage by these xenobiotics are described. PMID- 20009124 TI - [Angiogenesis and immune suppression: yin and yang of tumor progression?]. AB - Specialized variants of neoplastic cells that appear in tumors during cancer disease progression possess the ability to recruit certain kinds of hematopoietic and mesenchymal cells from the bone marrow or bloodstream. These tumor-recruited hematopoietic cells include monocytes, macrophages, granulocytes, mast and dendritic cells, as well as myeloblastic suppressor cells. Fibroblasts derived from undifferentiated mesenchymal cells are also recruited. Some of these cells (especially macrophages and fibroblasts) then undergo "education-like" phenotype reprogramming under the influence of the neoplastic cell population, resulting in the appearance of tumor-associated macrophages (TAM) and fibroblasts (CAF). Together with the extracellular matrix (ECM) as well with the remaining types of recruited cells, they contribute to the formation of a specific tumor microenvironment. Both the cells forming the tumor microenvironment and neoplastic cells engage in the two intimately linked processes of angiogenesis and immune suppression. The network of defective blood vessels formed during tumor angiogenesis and the resulting fluctuations in blood flow lead to under oxygenation of the surrounding neoplastic cells and have substantial impact on their metabolic profile. A number of processes triggered in these under oxygenated neoplastic cells appear to strongly favor further tumor progression. Such processes result in lower oxygen demand, enhanced angiogenesis, and epithelial-mesenchymal transition, owing to which the neoplastic cells acquire the ability to translocate. Under-oxygenation also leads to augmented genetic instability of the neoplastic cells. The tumor environment-forming cells also have their share in the establishment of an immunosuppressive environment which enables the neoplastic cells to escape immune surveillance. By providing a sophisticated milieu for the selection of increasingly malignant neoplastic cells (i.e. with proangiogenic and immunosuppressive phenotypes), the tumor microenvironment-forming cells substantially contribute to the progression of a neoplasm. Inhibited angiogenesis thus makes an immune response, both nonspecific and specific, possible. The remarks presented here may prove helpful in devising novel anticancer strategies involving antiangiogenic in combination with immunomodulatory drugs. PMID- 20009126 TI - Reversibility of liver fibrosis. AB - Liver cirrhosis is a major cause of morbidity and mortality worldwide and has very limited therapeutic options. Regardless of the aetiology, hepatic fibrosis is a characteristic feature of chronic liver disease. Our knowledge regarding the pathogenesis of this scarring has grown exponentially in the past 25 years. It has now clear that this is a highly dynamic process and the long-held dogma that it is irreversible and relentlessly progressive is now being challenged. In this review, we will summarise the key pathogenic mechanisms at play and will focus on the evidence demonstrating that liver fibrosis is reversible in humans and animal models. In particular, we will examine the role of hepatic stellate cells, MMPs, TIMPs and macrophages in this process. Finally, we will discuss some of the studies aimed to therapeutically target the resolution of fibrosis and their potential for translation into a badly-needed treatment modality in the clinical setting. PMID- 20009127 TI - Antiviral therapy in patients with HCV-cirrhosis. AB - The main cause of liver cirrhosis and liver cancer in the western world is Hepatitis C virus (HCV) infection. Liver transplantation is the only effective treatment once the disease is decompensated. In viremic patients who undergo transplantation, disease recurrence is universal resulting in the development of a new cirrhosis in about one third of the patients after 5 to 10 years of follow up. Initiation of the antiviral treatment with Peg-IFN and ribavirin prior to transplantation may prevent HCV recurrence if a sustained viral response (SVR) is achieved. Moreover, it might even be possible to achieve an improvement of the liver function degree so that transplantation may be differed. There are few studies that assess the efficacy and safety of the antiviral treatment in the cirrhotic setting. Available information shows SVR rates between 20 and 40%, lower with decompensated disease. The need for treatment withdrawal and dose reductions is significant in this setting. Cytopenias are one of the most frequent adverse effects; hematopoietic growth factors have shown to increase patient compliance, but it is still unclear whether they result in greater SVR. In addition, an increased risk of bacterial infections has been recently described, with a recommendation to use prophylactic therapy during antiviral treatment. In conclusion, antiviral therapy is an option for cirrhotic patients who have a good liver function but should not be recommended in patients with Child-Pugh-Turcotte class C, due to a high risk of severe complications. PMID- 20009128 TI - Adult living donor versus deceased donor liver transplantation: a 10-year prospective single center experience. AB - It has been 4 years since the first, long-term (> 3 years) prospective comparison of adult-to-adult living donor liver transplantation (A2ALLTx) to adult deceased donor liver transplantation (ADDLTx) was reported. In this follow up, prospective, IRB approved, 10-year comparison of A2ALLTx to ADDLTx we expand on our initial observations. This data includes: age, gender, ethnicity, primary liver disease, waiting time, pretransplant CTP/MELD score, cold ischemia time (CIT), perioperative mortality, acute and chronic rejection, graft and patient survival, charges and post-transplant complications. In 10 years, 465 ADDLTx (81.3%) and 107 A2ALLTx (18.7%) were performed at VCUHS. Hepatitis C virus (HCV) was the most common reason for transplantation in both groups (54.5% vs. 48.2%). Data regarding overall patient and graft survival and retransplantation rates were similar. Comparison of patient/graft survivals, retransplantation rates in patients with and without HCV were not statistically different. A2ALLTx patients had less acute rejection (9.6% vs. 21.7%) and more biliary complications (27.1% vs. 17.6%). In conclusion, A2ALLTx is as durable a liver replacement technique as the ADDLTx. Patients with A2ALLTx were younger, had lower MELD scores, less acute rejection and similar histological HCV recurrence. Biliary complications were more common in A2ALLTx but were not associated with increased graft loss compared to ADDLTx. PMID- 20009129 TI - Predicting 6-week mortality after acute variceal bleeding: role of Classification and Regression Tree analysis. AB - BACKGROUND: Available prognostic scores for mortality after acute variceal bleeding are mainly based on logistic regression analysis but may have some limitations that can restrict their clinical value. AIMS: To assess the efficacy of a novel prognostic approach based on Classification and Regression Tree -CART- analysis to common easy-to-use models (MELD and Child-Pugh) for predicting 6-week mortality in patients with variceal bleeding. METHODS: Sixty consecutive cirrhotic patients with acute variceal bleeding. CART analysis, MELD and Child Pugh scores were performed to assess 6-week mortality. Receiver operating characteristic (ROC) curves were constructed to evaluate the predictive performance of the models. RESULTS: Six-week rebleeding and mortality were 30% and 22%, respectively. Child-Pugh and MELD scores were clinically relevant for predicting 6 weeks mortality. CART analysis provided a simple algorithm based on just three bedside-available variables (albumin, bilirubin and in-hospital rebleeding), allowing accurate discrimination of two distinct prognostic subgroups with 3% and 80% mortality rates. All MELD, Child-Pugh and CART models showed excellent and comparable predictive accuracy, with areas under the ROC curves (AUROC) of 0.88, 0.84 and 0.91, respectively. CONCLUSIONS: A simple CART algorithm combining albumin, bilirubin and in-hospital rebleeding allows an accurate predictive assessment of 6-week mortality after acute variceal bleeding. PMID- 20009130 TI - Clinical and basal aspects of anemia during antiviral therapy for hepatitis C. AB - BACKGROUND AND RATIONALE: Anemia is a major side effect of combination therapy for chronic hepatitis C. In this study, severity, potential risk factors for and potential underlying mechanisms of anemia were evaluated. PATIENTS AND METHODS: 44 chronic hepatitis C patients on interferon-ribavirin treatment were included. Anemia-related parameters were measured before and during treatment. Potential changes in membrane phospholipids composition of erythrocytes of patients on anti viral treatment and potentially increased erythrocyte susceptibility to osmotic or bile salt induced stress were explored. RESULTS: Anemia was almost universal during treatment, with evidence of hemolysis. Decrease of Hb after six months of therapy was 2.1 +/- 0.1 mmol/L (range -0.6-4.1). Higher pre-treatment Hb, highest ribavirin dose (15-17.5 mg/kg) and lower pre-treatment platelet level were independent risk factors for decrease of Hb. Serum erythropoietin levels increased during treatment with negative correlation to Hb levels at week 12 (r = -0.70, p = 0.002) and 24 (r = -0.72, p = 0.002). Erythrocyte membrane phospholipid composition did not differ between anemic patients and healthy controls. Also, resistance to osmotic or bile salt induced stress was normal in anemic patients. Phosphatidylserine exposure at the outer membrane leaflet did not change upon 24 hrs ex vivo incubation with pharmacological ribavirin concentration. CONCLUSIONS: Anemia is almost universal during anti-HCV treatment. The extent of anemia correlates with pre-treatment levels of thrombocytes and Hb and with high ribavirin dosing. Although we found hemolysis as contributing factor, our data do not indicate that altered membrane phospholipids composition is an important factor in pathogenesis of anemia. PMID- 20009131 TI - Platelet count/spleen diameter ratio for non-invasive prediction of high risk esophageal varices in cirrhotic patients. AB - BACKGROUND AND OBJECTIVE: Prophylaxis therapy is indicated in cirrhotic patients with large esophageal varices or small varices with red wale signs (high risk esophageal varices; HREV). Endoscopic surveillance to detect HREV is currently recommended. The objective of this study is to identify non-invasive predictors of HREV in cirrhotic patients. DESIGN AND METHODS: Adult cirrhotic patients without previous variceal bleeding were prospectively included. All patients underwent a complete biochemical workup, upper digestive endoscopy, and ultrasonographic measurement of spleen bipolar diameter. Platelet count/spleen diameter ratio (PC/SD) was calculated for all patients. The association of these variables with the presence of HREV in upper endoscopy was tested using univariate and multivariate analysis. Receiver operating characteristic (ROC) curves were constructed for variables associated with HREV. RESULTS: Sixty-seven patients were included. The prevalence rate of HREV was 50%. Age, gender (female), platelet count, spleen diameter, PC/SD ratio, total bilirrubin, prothrombin activity (INR), Child-Pugh score, clinical and ultrasonographic ascites were significantly associated with presence of HREV in univariate analysis. Age and PC/SD ratio were the parameters independently associated with HREV in a multivariate analysis, with OR 8.81 (CI 95%: 1.7-44.9) and OR 11.21 (CI 95%: 2.8-44.6) respectively. A PC/SD ratio cut-off value under 830.8 predicted HREV with 76.9% sensitivity, 74.2% specificity and 77.8% negative predictive value (ROC curve area: 0.78). CONCLUSIONS: The PC/SD ratio was significantly associated with HREV, but with suboptimal sensitivity and specificity. Therefore, the results of this study do not support the routine clinical use of PC/SD ratio for screening of HREV. PMID- 20009132 TI - Altered clot kinetics in patients with non-alcoholic fatty liver disease. AB - BACKGROUND: Emerging evidence has linked the presence of non-alcoholic fatty liver disease (NAFLD) with an increased risk for cardiovascular events. We hypothesised that altered clot kinetics and platelet function may contribute to this increased risk. This study compared whole blood clotting kinetics in patients with 1) non-cirrhotic NAFLD (n = 28) and 2) healthy control subjects (n = 22). METHODS: Clotting kinetics were assessed in whole blood using thromboelastography (TEG) and assessed for correlations with cardiovascular risk factors. RESULTS: Clot kinetics in patients with NAFLD showed significantly stronger clot development (maximum amplitude (MA); 58.3 +/- 6.3 mm vs. 52.0 +/- 10.1 mm, p = 0.01) and reduced clot lysis in the presence of thrombin (35 +/- 30% vs. 51 +/- 26% clot lysis 30 minutes after MA, p = 0.03) compared to control subjects. Clot strength was independently positively associated with body mass index in NAFLD, but not in control subjects. There was a greater platelet contribution to clot strength in patients with NAFLD compared to controls despite similar platelet counts. There was no association between clot kinetics and features of the metabolic syndrome or presence of type 2 diabetes. CONCLUSION: Patients with NAFLD have disturbances in ex-vivo clot kinetics including increased clot strength and clots that are more resistant to thrombin-stimulated lysis. PMID- 20009133 TI - Patterns of viral load in chronic hepatitis B patients in Brazil and their association with ALT levels and HBeAg status. AB - Serum hepatitis B virus (HBV) DNA level is a predictor of the development of cirrhosis and hepatocellular carcinoma in chronic hepatitis B patients. Nevertheless, the distribution of viral load levels in chronic HBV patients in Brazil has yet to be described. This cross-sectional study included 564 participants selected in nine Brazilian cities located in four of the five regions of the country using the database of a medical diagnostics company. Admission criteria included hepatitis B surface antigen seropositivity, availability of HBV viral load samples and age >or=18 years. Males comprised 64.5% of the study population. Mean age was 43.7 years. Most individuals (62.1%) were seronegative for the hepatitis B e antigen (HBeAg). Median serum ALT level was 34 U/L. In 58.5% of the patients HBV-DNA levels ranged from 300 to 99,999 copies/mL; however, in 21.6% levels were undetectable. Median HBV-DNA level was 2,351 copies/mL. Over 60% of the patients who tested negative for HBeAg and in whom ALT level was less than 1.5 times the upper limit of the normal range had HBV-DNA levels > 2,000 IU/mL, which has been considered a cut-off point for indicating a liver biopsy and/or treatment. In conclusion, HBV-DNA level identified a significant proportion of Brazilian individuals with chronic hepatitis B at risk of disease progression. Furthermore, this tool enables those individuals with high HBV-DNA levels who are susceptible to disease progression to be identified among patients with normal or slightly elevated ALT. PMID- 20009134 TI - NASH and cryptogenic cirrhosis: a histological analysis. AB - INTRODUCTION: Epidemiological studies indicate that nonalcoholic steatohepatitis (NASH) is a common cause of cirrhosis described as 'cryptogenic'. To address this from a histological perspective and to examine the significance of residual histological findings as an indication of prior NASH, we looked back at biopsies in patients who presented with cirrhosis without sufficient histological features to diagnose NASH but who had prior histologically confirmed non-cirrhotic NASH. METHODS: Seven patients were identified with biopsy pairs showing non-specific (cryptogenic) cirrhosis in the latest specimen and a prior biopsy showing non cirrhotic NASH. Using an expanded NASH-CRN system scored blindly by light microscopy, we compared the early and late biopsies to each other and to a cohort of 13 patients with cirrhosis due to hepatitis C without co-existing metabolic syndrome. RESULTS: Macrosteatosis, although uniformly present in the non cirrhotic NASH specimens, declined in the late stage cirrhotic NASH specimens and was not useful in the distinction of late cirrhotic NASH from cirrhotic viral hepatitis. However, the presence of ballooned cells, Mallory-Denk bodies, and megamitochondria and the absence of apoptotic bodies were significantly different in late stage cirrhotic NASH compared to cirrhosis due to hepatitis C. CONCLUSIONS: Histologically advanced NASH presenting as non-specific or cryptogenic cirrhosis has residual changes which are consistent with prior steatohepatitis but which differ from cirrhosis due to hepatitis C. These results provide histological support for the more established epidemiological associations of NASH with cryptogenic cirrhosis and for criteria used in several proposed classifications of cryptogenic cirrhosis. PMID- 20009135 TI - Early treatment with N-acetylcysteine in children with acute liver failure secondary to hepatitis A. AB - INTRODUCTION: Hepatitis A virus can evolve to acute liver failure with a fatal outcome if it is not reversed. OBJECTIVE: We describe the clinical course of 12 children who presented with hepatitis A acute liver failure and received treatment with oral N-acetylcysteine (NAC). MATERIALS AND METHODS: Of the seventy two patients with viral hepatitis A, 12 patients who had acute hepatic failure were included. The variables evaluated were age, sex, duration of clinical features prior to hospitalization, signs and symptoms, laboratory parameters [alanine aminotransferase (ALT), aspartate aminotransferase (AST), prothrombin time (PT), partial thromboplastin time (PTT), internal normalization ratio and ammonia], treatment (oral NAC 100 mg/kg/day, lactulose, neomycin and general measures) and clinical course during hospitalization. RESULTS: Six males and six females were included. School-aged and adolescent children predominated. All presented with jaundice, nausea, vomiting and hepatomegaly. Two had stage 2 neurological signs as per the West-Haven scale. All had altered laboratory parameters. All received NAC, six patients for a week and the remaining six for 9 36 days. Treatment was not ceased until patients showed clinical and laboratory improvement. All data were analyzed using both student's t test and Wilcoxon signed rank with alpha = 0.05, the ALT with P = 0.0003 and 0.005, AST with P = 0.0001 and 0.0005, PT with P = 0.0237 and 0.0005, PTT with P = 0.0515 and 0.0039, ammonia with P = 0.0197 and 0.0015 and direct bilirubin with P = 0.0190 and 0.068. There was good tolerance to medications and a satisfactory clinical course. DISCUSSION: The use of oral NAC appears to be an effective therapeutic alternative for hepatitis A-induced liver failure if it is offered appropriately. It can modify the clinical course to a favorable one and prevent the fatal outcome of hepatic encephalopathy. PMID- 20009136 TI - The safety and feasibility of large volume paracentesis performed by an experienced nurse practitioner. AB - Due to time constraints, fewer physicians are performing large volume paracentesis (LVP) resulting in a longer wait time and more emergency department (ED) and hospital admissions. At our institution, after initial supervision, a certified nurse practitioner (NP) has independently performed LVP in a dedicated cirrhosis clinic. The purpose of our study was to evaluate the feasibility and safety of LVP performed by a NP. A retrospective review of patients undergoing LVP between January 2003 and May 2007 was performed. Baseline patient information and the practitioner performing LVP (physician or NP) were recorded. Complications including post paracentesis hypotension, bleeding, local leakage of ascitic fluid, infection, perforation, and death were compared between the two groups. A total of 245 procedures in 41 patients were performed by a single NP, and 244 in 43 patients by physicians. Baseline characteristics of patients undergoing LVP were similar in two groups. Alcohol was the most common etiology of cirrhosis (46% in NP and 51% in physician group) followed by a combination of alcohol plus HCV (37% in NP and 28% in physician group). There was similar distribution of Childs class B and C patients in the two groups, as well as average MELD score. Total volume of ascites removed, number of needle attempts, and complications including post paracentesis hypotension, local leakage of ascitic fluid, bleeding, infection, and death were not statistically different between the two groups. Our study shows no difference between physician and NP performance of LVP and complication rates. LVP performed by a NP is feasible and has acceptable rate of complications. PMID- 20009137 TI - Hepatoprotective effect of pentoxifylline against D-galactosamine-induced hepatotoxicity in rats. AB - The present study was conducted to investigate effect of pentoxifylline (PTX) on acute liver injury caused by galactosamine (D-Gal) in rats and the underlying mechanism involved in this setting. Moreover, we attempted to compare its effect to the well-established hepatoprotective agent, silymarin (SYM). The rats were randomly assigned 5 groups, control, PTX-treated (100 mg/kg, 3 weeks), SYM treated (100 mg/kg, 3 weeks) and their combination. Hepatic injury was induced by intraperitoneal single dose injection of D-Gal (800 mg/kg). Hepatic functions parameters, including serum albumin and alkaline phosphatase (ALP) levels were determined. Antioxidants enzyme activities such as superoxide dismutase (SOD), catalase (CAT) as well as lipid peroxides and hepatic total nitrites were measured. Besides, histopathological examination was also performed using portions of liver tissues. Results showed that the liver injury induced by D-Gal was improved in the three pretreated groups to variable extents. Pretreatment with PTX prevented D-Gal-induced reduction of antioxidant enzyme activities, SOD and CAT, and attenuated the elevated malonaldahyde (MDA) level in hepatic tissue as marker of lipid peroxidation. In addition, pretreatment with PTX resulted in an increase in hepatic triglycerides, normalization of nitric oxide level, and lowering serum ALP activity as well as inhibited the decreased serum albumin level caused by D-Gal. These biochemical changes were reflected on attenuation the structural alterations of the liver integrity. Collectively, our data suggest that PTX exhibits a potential hepatoprotective effect against D-Gal-induced hepatotoxicity and this effect might be attributed to its antioxidant properties. PMID- 20009138 TI - Acute treatment of constant darkness increases the efficiency of ATP synthase in rat liver mitochondria. AB - The circadian oscillations of many physiological processes provide an endogenous temporal program for the adaptive synchronization of mammals to the fluctuating external world. The lack of exposure to light causes the circadian system to undergo a process of dark adaptation similar to dark adaptation in the visual system. The aim of the present work was investigate the effect of acute treatment of constant darkness on mitochondrial ATP synthase activities and membrane fluidity in liver from male rat. We found that ATP synthase activity was not changed by the treatment. However ATPase activity and membrane fluidity were significantly diminished and pH gradient driven by ATP hydrolysis was incremented, in comparison from samples from rats kept on normal light/dark cycles. Additionally, the treatment of constant darkness diminishes the passive proton permeability of the inner mitochondrial membrane. In conclusion constant darkness induces a more efficient coupling between proton transport and catalysis, and increment the efficiency of the enzyme because the ratio of ATP synthase/ATPase activity was higher. These results exhibited the physiological adaptation of liver mitochondria to acute treatment of constant darkness in order to satisfy the cellular energy demand. PMID- 20009139 TI - A giant hepatic hemangioma treated successfully with hepatic enucleation. PMID- 20009140 TI - De-novo cholangiocarcinoma in native common bile duct remnant following OLT for primary sclerosing cholangitis. AB - Primary sclerosing cholangitis (PSC) is a chronic, progressive, inflammatory and obstructive disease of the intra- and extra-hepatic bile ducts of unknown etiology. Currently, orthotopic liver transplantation (OLT) is the only definitive treatment for PSC-related end-stage liver disease. However, PSC has been known to recur in the grafted liver. Roux-en-Y hepaticojejunostomy is more commonly performed than choledochocholedochostomy for PSC, although choledochocholedochostomy has been found to be safe and efficacious for PSC if the distal common bile duct is uninvolved at the time of OLT. Our case is unique in that it describes a patient who developed de-novo cholangiocarcinoma in the remnant portion of the native common bile duct six years after OLT with choledochocholedochostomy for PSC-associated end-stage liver disease without having PSC recurrence. In conclusion, our case report indicates that choledochocholedochostomy may not be desirable in PSC due to an increased risk of developing cholangiocarcinoma in the native common bile duct. This risk exists as well with a Roux-en-Y hepaticojejunostomy in the remaining intra-duodenal and intra-pancreatic biliary epithelium, although in theory to a lesser extent. Therefore, the risk of developing cholangiocarcinoma in the recipient common bile duct can only be completely eliminated by performing a Whipple procedure at the time of OLT. PMID- 20009141 TI - Spontaneous rupture of hepatic metastasis from choriocarcinoma: a review of imaging and management. AB - Spontaneous rupture of hepatic metastasis causing hemoperitoneum is a rare entity. Ruptured hepatic metastasis has typical imaging findings on biphasic CT which may help in clinching the diagnosis. We present a case of rupture hepatic metastasis from choriocarcinoma in a young-female patient who was managed by transcatheter hepatic artery embolization. A brief review of the imaging features and therapeutic options for the ruptured hepatic metastases is discussed along with the case. PMID- 20009142 TI - Budd Chiari Syndrome without evidence for thrombosis. Is it still a Budd Chiari? PMID- 20009143 TI - Noncirrhotic portal hypertension: another cause of liver disease in HIV patients. AB - Noncirrhotic portal hypertension (NCPH) represents a relatively infrequent group of conditions that causes portal hypertension in the absence of cirrhosis. An association between NCPH and patients infected with human immunodeficiency virus (HIV) has been reported. Six consecutive patients with HIV infection and NCPH were the subject of this series. Case histories, including medication lists, liver biopsy and laboratory data were reviewed. Age at diagnosis was 43 +/- 3 years (range, 37-47). Liver disease was diagnosed 12 +/- 4 years (range, 8-18) after initiation of antiretroviral therapy (ART). All patients developed esophageal varices, 5 patients presented at least one bleeding episode and 2 required TIPS. Serum liver tests showed a mean total bilirubin of 1.4 +/- .7 mg/dL (range, .5-2.5) and INR was 1.2 +/- .14 (range, 1.0-1.4). CD4 count was 326 +/- 124 cells/mL (range, 198-467) and all patients presented HIV viral load < 75 copes/mL. Didanosine (ddl) was the most common ART drug being used by 4 patients. Portal vein thrombosis was diagnosed in 2 patients. Hepatic portal sclerosis (HPS) alone was observed in 1 patient, nodular regenerative hyperplasia (NRH) alone in 2 patients and combined HPS/NRH in 3 patients. In conclusion, NCPH should be included in the differential diagnosis of HIV-individuals presenting with clinical manifestations of portal hypertension and well preserved liver synthetic function. Prolonged exposure to ART, specially ddl, can play a pathogenic role. Rarely, liver synthetic function is sufficiently severe to warrant liver transplantation. PMID- 20009144 TI - Late acute celiac and hepatic artery thrombosis with portal vein thrombosis resulting in hepatic infarction 12 years post orthotopic liver transplantation. AB - Hepatic artery thrombosis (HAT) is relatively infrequent, but possibly a devastating complication of orthotopic liver transplantation (OLT). It often requires urgent retransplantation. Two main forms of HAT are recognized as early and late HAT (diagnosis within or after 30 days following LT). Early HAT typically results in graft failure. Late HAT features biliary obstruction, cholangitis, and hepatic abscess formation. We report here the case of a patient of Wilson's disease who presented twelve years post-liver transplant symptoms typical of acute HAT and hepatic infarction. On diagnostic imaging, celiac axis and hepatic artery were thrombosed, resulting in ischemic necrosis of the left hepatic lobe. The resulting sepsis and transient hepatic insufficiency were managed conservatively, and repeat OLT was avoided. The patient remains stable more than one year later. To the best of our knowledge this case report is unique in the literature for the unusually long interval between OLT and late acute HAT, as well as celiac and portal vein occlusion. The acute presentation of sub massive hepatic necrosis is also uncharacteristic of late HAT and more typical of acute HAT. This report describes our experience in managing this and a literature review of the topic. PMID- 20009146 TI - Ciprofloxacin-induced acute cholestatic hepatitis. PMID- 20009147 TI - Pentoxifylline and prednisolone in severe alcoholic hepatitis. PMID- 20009148 TI - Intravenous administration of N-acetylcysteine reduces plasma total homocysteine levels in renal transplant recipients. AB - BACKGROUND: Hyperhomocysteinemia occurs in approximately 60-70% of renal transplant recipients and is associated with increased risk of cardiovascular events, mortality and kidney allograft loss. In normal subjects N-acetylcysteine (NAC) given either orally or intravenously markedly reduces plasma total homocysteine (tHcy) level. In cardiac transplant recipients it was reported, that oral treatment with NAC does not affect Hcy levels. We have therefore, investigated the effect of intravenous NAC on plasma tHcy levels in renal transplant recipients. MATERIAL/METHODS: Eleven renal transplant recipients who had normal plasma levels of vitamin B12 and folic acid, were treated with intravenous NAC or placebo in a crossover manner. RESULTS: Intravenous administration of NAC significantly reduced plasma tHcy (p=0.0008). Decrease in tHcy was related to its initial concentration. CONCLUSIONS: Intravenous NAC profoundly reduces tHcy level in renal transplant recipients. Further research is needed to establish the effect of orally administered NAC on plasma homocysteine concentration in this clinical condition. PMID- 20009150 TI - Causes of re-hospitalization in different post kidney transplantation periods. AB - BACKGROUND: Post kidney transplantation Re-admissions are focused because they are costly and cause morbidity, or may end with unsatisfactory endpoints namely graft loss or death. We compared the pattern, outcome and cost of re-admissions in different post-kidney transplantation periods. MATERIAL/METHODS: In a retrospective study, 562 consecutive re-admissions of kidney recipients categorized to early (during first 6 months; n=278); intermediate (6-24 months; n=115); and late (24 months and afterwards, n=169) hospitalizations. Primary outcome measures included hospitalization pattern (cause and length of hospital stay), and secondary outcome measure were assessed (mortality and graft loss during hospitalization) and costs. RESULTS: The causes of rehospitalization were surgical complication (84 percent), infection (51 percent), graft rejection (45 percent), and malignancy (0.6 percent), in early phase, graft rejection (44 percent), infection (42 percent), surgical complication (13 percent), and malignancy (5 percent), in intermediate phase, and graft rejection (45 percent), infection (39 percent), surgical complication (3 percent), and malignancy (0.06 percent), in late phase. So, infections and surgical complications showed a decreasing trend from early to late post transplant phase, while malignancies showed a peak in intermediate phase. The length of hospital stay (12+/-11, 10+/ 10, 9+/-7, p=0.001) and hospitalization charges (708+/-36, 468+/-333, 413+/-262 united states Dollars, p=0.035) were significantly higher in the early post transplant phase. Mortality (p=0.755) and graft loss during hospitalization (p=0.246) remained the same in all time intervals. CONCLUSIONS: Early post-kidney transplantation phase, with a higher risk of infections and surgical complications, health care system experience longer and more costly hospitalizations. PMID- 20009149 TI - Factors determining changes in concentrations of pro-inflammatory markers in blood serum in the initial period after kidney transplantation from dead donor. AB - BACKGROUND: The main aim of this paper is to determine scope of changes in concentration of selected pro-inflammatory factors in blood serum in the initial period after the kidney transplantation from the brain death donor and attempt of establishing which of them are connected with delayed function of the organ, initial condition of both the donor and the recipient. MATERIAL/METHODS: The prospective examination carried out in group of 20 patients that underwent kidney transplantation from 10 donors diagnosed brain death. Blood samples were drawn before the transplantation procedure, 4 hours after procedure and in 4(th) day after the procedure. The blood serum analysis included: interleukin-6 (IL-6), tumor necrosis factor-alpha, interleukin-1beta (IL-1b) and C-reactive protein (CRP). RESULTS: In perioperative period the dynamic changes of all observed markers were noticed, especially in 4 th hour after the transplantation procedure. Generally, apart from TNFalpha, their concentration in blood serum was raising (the most significantly for IL-6) and then in 4(th) day reached the level equal to or lower than before transplantation. CONCLUSIONS: The delayed graft function is accompanied by high CRP level in donors before grafting and sustaining rise of IL-1b content in blood serum in 4(th) day after the transplantation procedure. The IL-6 content in this period revealed similar tendency in recipients' pairs that have been given the kidney from one donor, reflecting the condition of an organ that has been transplanted. PMID- 20009151 TI - Improvement of renal transplantation outcome through matching donors and recipients. AB - BACKGROUND: To investigate the effects of age and gender matching on patient and graft survival in living unrelated kidney transplantation. MATERIAL/METHODS: All 2649 first-time kidney transplanted cases who had received their graft from a living unrelated donor in Baqiyatallah Hospital (Tehran, Iran) were enrolled (1992-2005). Based on the age and gender matching state of the donors (D) and recipients (R), the recipients were divided into four age-match (A) and four gender-match (G) groups. Age-match groups included A(1) (R< or =40, D< or =40, n=1483), A(2) (R>40, D< or =40, n=1044), A(3) (R< or =40, D>40, n=82) and A(4) (R>40, D>40, n=40). Gender-match groups comprised G(1) (R: female, D: female, n=209), G(2) (R: male, D: male, n=1428), G(3) (R: female, D: male, n=768) and G(4) (R: male, D: female, n=244). Using Kaplan-Meier method, 6-month, to 5-year graft/patient survival rates were determined for different patient groups. Survival curves were compared using log rank test after stratification. RESULTS: Male recipients living with a female donor's kidney had a shorter survival compared to both the males having received a male kidney and the females having received a female kidney. Graft survival also showed a marginally significant difference and was shorter among the males with a female kidney graft compared to the males living with a male kidney graft. In contrast, donor's sex caused no difference in patient or graft survival among female recipients. When survival curves of age-match groups were compared, both graft and patient survival times were significantly shorter among the younger patients having a kidney graft received from an older donor. Patient survival was also shorter among old recipients having received an old kidney compared with old patients having a kidney graft from a young donor. Though graft survival among the old patients with old grafts was also shorter than in those with young grafts, the difference was not significant. When the subjects were stratified by donors' age, the results showed that old patients with young kidneys survived shorter than young patients with young kidneys. CONCLUSIONS: Better living unrelated renal transplantation outcome is expected with younger kidney grafts (i.e. donor < or =40 yr) and avoiding grafting female kidneys into male recipients. PMID- 20009152 TI - A single-center experience of renal transplantation in elderly patients. AB - BACKGROUND: We retrospectively reviewed the results of renal transplantation in patients over 60 years of age at our center. MATERIAL/METHODS: A retrospective study was conducted of 212 Bahraini patients receiving renal transplants from January 1979 to December 2007. All medical records were reviewed for demographic data, graft function and survival. Patient and graft survival was compared for patients above and below the age of 60. RESULTS: Seventeen patients >60 years with a mean age of 64.1+/-3.6 years at the time of transplantation. Diabetic nephropathy (52%) was the most common causes of end-stage renal disease. Mean donor age was 26+/-6 years and most of them were unrelated (82%). Of the elderly patients, 4 died: 3 with a functioning graft, 1 within one year of transplantation. Cardiovascular causes (3 patients, 75%) were the most common cause of death. Causes of graft loss were death with a functioning graft (4) and chronic rejection (1). Cox's proportional-hazards regression analysis showed on univariate analysis that pre-transplant hypertension, diabetes mellitus and vascular surgery (CABG) before transplant significantly affected the dependent variable of graft loss. Multivariate analysis did not show these variables to be significant. Kaplan Meier patient survival curves showed statistically significant differences between study (>60 years) and control (18-59 years) patients (p=0.04) at 10 year. CONCLUSIONS: We conclude that Patients older than 60 yr of age can be transplanted safely and successfully, especially when they are properly screened for the presence of significant risk factors such as severe cardiovascular disease and diabetes. PMID- 20009153 TI - Risk factors for post operative pulmonary complications and mortality in cardiac transplantation patients. AB - BACKGROUND: Pulmonary disorders are common in patients with advanced heart failure (HF). In this study we sought to evaluate the findings detected on pre and postoperative assessment of heart failure patients who underwent orthotopic heart transplantation (OHT). MATERIAL/METHODS: The records of 30 patients with advanced HF who underwent OHT at our center were evaluated. Clinical and demographic characteristics, laboratory and radiological findings were noted. Pulmonary function tests (PFTs) and cardiopulmonary exercise testing (CPET) results were obtained. RESULTS: The mean age was 31.0+/-16.8 years and 7 were female. History of smoking was present in 15 (50%) patients. PFTs revealed normal findings in 50%, obstructive pattern in 26.7%, restrictive in 16.7% and mixed pattern in 6.6% of the patients. CPET was performed to 17 patients. Mean peak VO2 of the patients was 11.5+/-4.9 ml/kg/min and mean VO2 at anaerobic threshold was 10.9+/-3.5 ml/kg/min. The mean left ventricular ejection fraction on the postoperative first week was 46+/-11%. The mean extubation time was 80.4 hours. Postoperative pulmonary complications were developed in 11 (36.7%) patients. Postoperative pneumonia was observed in 5 (16.7%) cases. Thoracentesis was performed to 7 patients with large pleural effusion, which was transudative in 5 and exudative in 2 patients. Thoracentesis culture was positive in 2 cases. Mortality was observed in 7 patients. CPET and PFTs results were found to have no significant effect on postoperative pulmonary complications and mortality (P>0.05). CONCLUSIONS: These findings indicate that pulmonary disorders are frequent in patients with advanced heart failure waiting heart transplantation, and following heart transplantation pulmonary complications are common. Further prospective researches with larger patient numbers are needed to determine risk factors for postoperative pulmonary complications in patients undergoing heart transplantation. PMID- 20009154 TI - Safety of mycophenolate mofetil monotherapy in patients after liver transplantation. AB - BACKGROUND: Due to the nephrotoxicity of calcineurin inhibitors (CNIs) mycophenolate mofetil (MMF) has become a promising immunosuppressive alternative in recent years. In several studies an improvement of renal function could be reached by introduction of MMF and withdrawal of CNIs but there is few known about the long-term effects of MMF on liver-, kidney- and bone marrow function. MATERIAL AND METHODS: We therefore retrospectively reviewed 123 adult liver transplant patients who were successfully converted to MMF monotherapy without any acute rejection within the first three months of MMF monotherapy. RESULTS: Serum levels of liver enzymes as well as white blood cells did not show any significant difference between the time of conversion and after 1 year but a significant reduction in serum creatinine could be evaluated (1.47 mg/dl vs. 1.54 mg/dl). In 59 patients liver biopsies were performed before and after introduction of MMF monotherapy. None displayed signs of chronic rejection during MMF monotherapy but there was a slight increase of the fat content of the liver (16.1% vs. 9.8%). CONCLUSIONS: In conclusion after successful conversion to MMF monotherapy this regime is safe and effective and the long-term effects of MMF monotherapy on the liver tissue are minor. Furthermore renal function can be improved without considerable effects on the bone marrow. PMID- 20009155 TI - Atypical calcineurin inhibitor-induced haemolytic uremic syndrome after liver transplantation. AB - BACKGROUND: A case of calcineurin inhibitor (CNI)--induced haemolytic uremic syndrome (HUS) after liver transplantation leading to irreversible renal failure is described. CASE REPORT: We present case history of 25-years old male after liver transplantation due to cryptogenic cirrhosis with prolonged worsening graft function, who developed HUS. Unsatisfactory graft function was the reason of performing numerous graft biopsies. Features of acute and chronic rejection (CR) of liver were histopathologically confirmed. Vanishing bile duct syndrome as manifestation of CR was stated and immunosuppressive regimen was intensified (tacrolimus placed cyclosporin). High blood levels of tacrolimus were maintained (approximately 20-22 ng/ml) on dose 3 mg twice a day. No clinical effect was observed. Renal failure was improving (serum creatinine was 3.3 mg/dl and eGFR was 24 ml/min/1.73 m(2)). After four months of maintaining high dose of tacrolimus patient was referred to our center in order to estimate indications for liver retransplantation. On admission severe haemolytic anaemia, thrombocytopenia and acute renal failure were detected. Atypical HUS probably related to CNI was diagnosed. Tacrolimus administration was discontinued. Blood and plasma transfusion as well as plasmapheresis were implemented. Haemolysis was limited, but renal function was not improved. Renal biopsy revealed features of irreversible nephropathy in course of thrombotic microangiopathy. Despite previously maintaining high dose of CNI, there were no signs of CNI nephrotoxicity. Patient required haemodialysis. Due to necessity of haemodialysis and worsening function of liver, patient was accepted to liver and kidney transplantation. CONCLUSIONS: High CNI blood concentration in patient after liver transplantation can be atypical cause of HUS and leads to irreversible renal failure. PMID- 20009156 TI - Aspergillus spondylodiscitis after multivisceral transplantation. AB - BACKGROUND: Although spondylodiscitis is rare, it is increasingly described in patients with compromised immunity due to malignancy, chemotherapy or immunosuppression. Typical pathogens are staphylococcus aureus and enterobacteria; fungal spondylodiscitis is uncommon. CASE REPORT: We present a case of aspergillus spondylodiscitis following pulmonary aspergillosis in a patient with multivisceral and kidney transplantation. Due to irreversible disc destruction, surgical restoration by autologous iliac crest graft was required in addition to intravenous antifungal therapy, which consisted of voriconazole, caspofungin and liposomal amphotericin B. CONCLUSIONS: Aspergillus spondylodiscitis is a diagnostic and therapeutic challenge, a combination of surgical debridement and antifungal therapy is inevitable to prevent rapid progression of invasive aspergillosis and neurological damage. PMID- 20009157 TI - Dose switch to another dosage form of Neoral increase the risk of medication error? AB - BACKGROUND: One of the most significant ways to avoid medication errors is to study the errors that have occurred in other institutions and to use the information to prevent similar accidents at other practice sites. CASE REPORT: We report a cyclosporine overdose that was caused, in part, by misinterpretation of the medication order of a transplanted patient. In transplantation regimen, a 15 mg BID dose of cyclosporine was supposed to be given as part of the immunosuppressive therapy. Unfortunately the patient received a total of 1500 mg but survived the overdose. CONCLUSIONS: This case should be considered in the development of strategies to prevent unfavorable outcomes resulting from such errors. PMID- 20009158 TI - Role of C3d and C4d complement fragments in the diagnostics of acute allograft rejection after transplantations. AB - Transplantation is a widely recognized method of treatment at the terminal stages of many renal, cardiac, hepatic and pulmonary diseases. Despite considerable advances in that field, graft rejection is still an important clinical problem. The reaction of the graft recipient to an organ presenting foreign antigens is dependent on complex immune mechanisms, involving both acquired and congenital immune responses. In most general terms, cellular and humoral immunity can be distinguished. The kidneys and the heart are the organs whose acute humoral rejection has been thoroughly investigated and defined, and the role of C4d and C3d fragments of the complement system has been confirmed by numerous studies. The studies concerning C4d and C3d expression in patients with acute humoral lung and liver rejection conducted to date have given contradictory results. Some of them confirm, while others fail to confirm, the correlation between their increased expression and AHR. From the practical point of view, C4d and C3d could be used in liver transplantology for differential diagnostics of acute graft rejection and recurrence of HCV infection. A few preliminary studies suggest the usefulness of these markers in the diagnostics of AMR and differentiation between both liver pathologies. There are only single reports concerning the role of C4d complement fragment in the diagnostics of acute rejection with a humoral component in case of small intestine grafts, as well as complex ones such as the hands and face, and their results suggests that these complement fragments are not important markers of acute rejection of these organs. PMID- 20009159 TI - Kidney dysfunction after non renal solid organ transplantation. AB - Chronic kidney disease developed after non renal solid organ transplantation in a significant issue. Evaluation of kidney function starting form a complete history and physical examination, then an accurate measurement of renal function and a urinalysis, followed by kidney imaging study should be performed prior to transplantation. Moreover, probability of reversibility of kidney dysfunction and the risk of progression to end stage renal disease should be assessed. During peri- and postoperative period hypotensive episodes should be avoided or at least minimized. Good blood pressure control (however, no guidelines for target blood pressure values for non solid organ transplants are available) as well as tight control of diabetes and hyperlipidemia should be implemented. Proteinuria, if present, should be attenuated by the use of therapeutic modalities (RAS blockade) to prevent or delay the progression of renal failure. The ability to identify CKD may allow early implementation of treatments that could arrest or delay the progression of renal damage, enable effective treatment of its complications, and reduce the risk of drug-induced nephrotoxicity. Kidney function should be monitored in nonrenal solid organ transplant recipients as an important risk factor comparable with diabetes for cardiovascular mortality and morbidity. In presence of existing nephron sparing immunosuppressive regimens, early detection of deteriorating kidney function gives the opportunity to tailor the treatment. A multidisciplinary approach for these patients including also a nephrologist should be considered. PMID- 20009160 TI - Late onset post transplantation lymphoproliferative disorders: analysis of international data from 5 studies. AB - BACKGROUND: Organ transplantation increases the risk of cancer including lymphomas through various and insidious mechanisms. In this study, we aimed to gather and analyze data of 5 international studies on late onset lymphoproliferative disorders after renal and liver transplantation. MATERIAL/METHODS: We conducted a comprehensive search for the available data by Pubmed and Google scholar search engines on post transplant lymphoproliferative disorders. A standard questionnaire was developed to collect data from different published studies. Finally, trustable data of a total of 5 publishes were included in the final analysis. We reviewed all their characteristics and various factors available which may affect outcome. We also compared outcome of patients with very late onset PTLD (>10 years) and late PTLDs. For data analysis we used SPSS v. 13.0. Kaplan-Meier Method was used for survival analysis and Students' t test and chi2 tests were used where appropriate. P<0.05 was defined as significant. RESULTS: overall 55 cases of PTLD from 5 international reports were entered into analysis. There were 32 (58%) males and 23 (42%) female patients. Mean age at diagnosis of PTLD was 39+/-12 years. The mean interval between transplantation and the diagnosis of PTLD was 86+/-54 whereas follow up time after diagnosis of PTLD was 34+/-99 months. Exteranodal involvement was 65%. There were no significant differences in outcomes of patients with late and very late onset PTLDs. Despite discontinuation or reduction of immunosuppressive agents, surgical therapy, chemotherapy, and radiotherapy, 55% of the patients died. CONCLUSION: Late onset PTLD has its highest relevance within the first year after diagnosis. Controlling infections in PTLD patients during this time period could have beneficial effects on patients' survival. Long post-transplant latency period, aggressive behavior and poor response to treatment necessitate early diagnosis and treatment of late onset post-transplant lymphomas in kidney transplant recipients. PMID- 20009161 TI - Autosomal dominant polycystic kidney disease and transplantation. AB - Autosomal dominant polycystic kidney disease (ADPKD) is an inherited disorder affecting 1 in 1,000 people and responsible for 10% of cases of the end stage renal disease (ESRD). Apart from renal manifestations, changes in other organs may be present. In the absence of contraindications, patients with ADPKD and ESRD should be referred to renal transplantation. The ADPKD patient may also need liver transplantation, or combined liver and kidney transplantation. Also, the patient with ADPKD may become a potential organ donor. The aim of our paper is to review the problems that the physicians deal with in ADPKD patients in pre- and post-transplant period. PMID- 20009162 TI - Cyclophosphamide dosage in pigs. PMID- 20009163 TI - Corrosion behaviors of TiO2 nanotube layers on titanium in Hank's solution. AB - It is well known that the growth of osteoblast cultured on titanium with nanotube layers can be significantly increased compared to unanodized surfaces. In the current study, the corrosion behavior of titanium with nanotube layers was studied in naturally aerated Hank's solution using open circuit potentials (OCP), electrochemical impedance spectroscopy (EIS) and potentiodynamic polarization tests. The electrochemical results indicated that TiO2 nanotube layers on titanium showed a better corrosion resistance in simulated biofluid than that of smooth-Ti. The OCP, corrosion rate (I(corr)), passive current density (I(pass)) and the oxygen evolution potential (E(o)) were significantly influenced by titanium oxide nanotube layers acquired by anodization. The anatase nanotube layer showed higher OCP and smaller current density than the amorphous nanotube layer. EIS analysis showed that the annealing had a significant effect on the corrosion resistance of the outer tube layer (R(t)), but little effect on the corrosion resistance of the inter-barrier layer (R(b)) for nanotube layers. The results suggested that titanium with TiO2 nanotube layers has an adequate electrochemical behavior for use as a dental implant material. PMID- 20009164 TI - Enhanced photoresponse of CdS/CMK-3 composite as a candidate for light-harvesting assembly. AB - Two typical carbon materials (ordered mesoporous carbon and carbon nanotube) were chosen as scaffolds in combination with semiconductor quantum dots (SQDs) for making light-harvesting assemblies. The effects of interfacial morphology on photoelectric performance of the carbon-based heterostructures have been investigated in detail. The enhanced photoresponse shows a strong dependence on the interfacial morphology as a result of direct interfacial contacts between SQDs and carbon materials, which plays a major role in increasing charge generation at the interface and transport pathways for photoinduced electron transfer. The methodology to enhance the photoresponse through tuning interfacial morphology proves to be a potent alternative in fabricating photochemical energy conversion systems. PMID- 20009165 TI - Observation of Peierls transition in nanowires (diameter approximately 130 nm) of the charge transfer molecule TTF-TCNQ synthesized by electric-field-directed growth. AB - We report the growth of nanowires of the charge transfer complex tetrathiafulvalene-tetracyanoquinodimethane (TTF-TCNQ) with diameters as low as 130 nm and show that such nanowires can show Peierls transitions at low temperatures. The wires of sub-micron length were grown between two prefabricated electrodes (with sub-micron gap) by vapor phase growth from a single source by applying an electric field between the electrodes during the growth process. The nanowires so grown show a charge transfer ratio approximately 0.57, which is close to that seen in bulk crystals. Below the transition the transport is strongly nonlinear and can be interpreted as originating from de-pinning of CDW that forms at the Peierls transition. PMID- 20009166 TI - Interaction of DNA/nuclear protein/polycation and the terplexes for gene delivery. AB - Nuclear transport of exogenous DNA is a major barrier to nonviral gene delivery that needs to be addressed in the design of new vectors. In this study, we prepared pDNA/HMGB1/PEG-PEI terplexes to promote nuclear import. HMGB1 in the terplexes was used to assist the transportation of pDNA into the nucleus of cells, since it contained nuclear localization signal (NLS); PEG chains were introduced to stabilize pDNA/vector terplexes and reduce the cytotoxicity. HMGB1/PEG-PEI combined vectors have been investigated specifically for their structure interaction by atomic force microscopy and circular dichroic spectroscopy. The results demonstrated that the HMGB1 molecule could bind with the pDNA chains, but not condense pDNA well. The PEG-PEI further compacted pDNA/HMGB1 complexes into nanosized spherical terplexes. The pDNA delivered by HMGB1/PEG-PEI combined vectors was significantly accumulated in the nucleus of cells, as observed by confocal laser scanning microscopy. The percentage of GFP transfected cells and VEGF protein expression level induced by HMGB1/PEG-PEI were 2.6-4.9-fold and 1.4-2.8-fold higher, respectively, than that of a common cationic polymer PEI 25 kDa. Therefore, the HMGB1/PEG-PEI combined vector could be used as a versatile vector for promoting exogenous DNA nuclear localization, thereby enhancing its expression. PMID- 20009167 TI - The optical properties of vertically aligned ZnO nanowires deposited using a dimethylzinc adduct. AB - The optical properties of zinc oxide nanowires are critically influenced by the growth process. Herein, we describe a metal-organic chemical vapour deposition (MOCVD) process for the growth of ZnO nanowires with improved optical properties. A tetrahydrofuran adduct is used to control the reactivity of dimethylzinc to enable this. Vertically aligned zinc oxide nanowires have been grown on Si(111) substrates by liquid injection MOCVD, using a solution of [Me(2)Zn(tetrahydrofuran)] in the presence of oxygen. The ZnO morphology becomes nanowire-like in a narrow temperature range centred about 500 degrees C. Above and below this temperature range, the ZnO is deposited in the form of polycrystalline films. The ZnO nanowires grow from a polycrystalline nucleation layer, with the (0002) c-axis parallel to the Si[111] substrate orientation. High resolution electron microscopy reveals a highly crystalline nanowire microstructure. Resonance enhanced ultraviolet Raman spectroscopy shows that the ratio of first- and second-order longitudinal optic modes is commensurate with electron-phonon coupling effects observed previously in ZnO nanostructures. Photoluminescence exhibits intense near band-edge emission with a full width at half-maximum of 110 meV at room temperature and shows negligible defect-related visible emission. PMID- 20009168 TI - Detailed modeling of the epitaxial growth of GaAs nanowires. AB - A detailed continuum model is presented for predicting the growth characteristics of GaAs nanowires during chemical beam epitaxy. The model describes the transport processes of Ga and As adatoms on the substrate and nanowire sidewalls, and through the nanoparticle and the nanowire-catalyst interface (NCI). The growth mechanisms of nanowires within the NCI are described using an extended step-flow kinetic model. The vapor-liquid-solid and vapor-solid-solid growth mechanisms are both described in the kinetic model. The growth rate of the nanowires, the surface and bulk concentrations of adatoms, and the role of transport processes of Ga and As adatoms during chemical beam epitaxy were investigated. The growth mechanisms of the nanowires were found to vary with increasing length of the nanowire. PMID- 20009169 TI - Investigation of resistive switching in Cu-doped HfO2 thin film for multilevel non-volatile memory applications. AB - In this paper, the resistive switching characteristics in a Cu/HfO(2):Cu/Pt sandwiched structure is investigated for multilevel non-volatile memory applications. The device shows excellent resistive switching performance, including good endurance, long retention time, fast operation speed and a large storage window (R(OFF)/R(ON)>10(7)). Based on the temperature-dependent test results, the formation of Cu conducting filaments is believed to be the reason for the resistance switching from the OFF state to the ON state. By integrating the resistive switching mechanism study and the device fabrication, different resistance values are achieved using different compliance currents in the program process. These resistance values can be easily distinguished in a large temperature range, and can be maintained over 10 years by extrapolating retention data at room temperature. The integrated experiment and mechanism studies set up the foundation for the development of high-performance multilevel RRAM. PMID- 20009170 TI - Selected immobilization of individual nanoparticles by spot-exposure electron beam-induced deposition. AB - The use of spot-exposure electron-beam-induced deposition (EBID) to immobilize targeted nanoparticles on a substrate is demonstrated, and investigated using experiment and simulation. Nanoparticles are secured in place through the build up of carbonaceous material that forms in the region between a particle and substrate when an energetic electron beam is focused onto the particle and projected through to the substrate. Material build-up directly affects the strength of adhesion to the surface, and can be controlled through electron dosage and beam energy. By selectively immobilizing specific particles within surface agglomerations and removing the excess, we illustrate the potential for spot-exposure EBID as a new technique for nanofabrication. PMID- 20009171 TI - Spontaneous self-organization of Cu2O/CuO core-shell nanowires from copper nanoparticles. AB - We report on the formation and spontaneous self-organization of Cu(2)O/CuO core shell nanowires from individual copper nanoparticles. The growth process is interpreted using the results of time-dependent in situ x-ray diffraction. High resolution transmission electron microscopy is used to observe the intermediate state of pearl-necklace-like aggregates that form a chain-like configuration of Cu(2)O nanoparticles intertwined into nanowires. The existence of an amorphous CuO shell is confirmed by the XANES technique and explained through an intensity simulation using a proposed core-shell nanowire model. PMID- 20009172 TI - The effect of catalysts and underlayer metals on the properties of PECVD-grown carbon nanostructures. AB - The growth behaviors and contact resistances of vertically aligned carbon nanotubes (CNTs) and carbon nanofibers (CNFs) grown on different underlayer metals are investigated. The average diameter, diameter distribution, density, growth rate and contact resistance exhibit strong correlation with the choice of catalyst/underlayer combination. These observations are analyzed in terms of interactions between the catalyst and the underlayer metal. The CNT via test structure has been designed and fabricated to make current-voltage measurements on single CNTs using a nanomanipulator under scanning electron microscopy (SEM) imaging. By analyzing the dependence of measured resistance on CNT diameter, the CNT-metal contact resistance can be extracted. The contact resistances between as grown CNTs and different underlayer metals are determined. Relationships between contact resistances and various combinations of catalysts and underlayer metals are investigated. PMID- 20009173 TI - Multi-walled carbon nanotube-based RF antennas. AB - A novel application that utilizes conductive patches composed of purified multi walled carbon nanotubes (MWCNTs) embedded in a sodium cholate composite thin film to create microstrip antennas operating in the microwave frequency regime is proposed. The MWCNTs are suspended in an adhesive solvent to form a conductive ink that is printed on flexible polymer substrates. The DC conductivity of the printed patches was measured by the four probe technique and the complex relative permittivity was measured by an Agilent E5071B probe. The commercial software package, CST Microwave Studio (MWS), was used to simulate the proposed antennas based on the measured constitutive parameters. An excellent agreement of less than 0.2% difference in resonant frequency is shown. Simulated and measured results were also compared against identical microstrip antennas that utilize copper conducting patches. The proposed MWCNT-based antennas demonstrate a 5.6% to 2.2% increase in bandwidth, with respect to their corresponding copper-based prototypes, without significant degradation in gain and/or far-field radiation patterns. PMID- 20009174 TI - Surface plasmon as a probe for melting of silver nanoparticles. AB - The temperature dependence of the surface plasmon energy and bandwidth for silver nanoparticles in the size range 8-30 nm embedded in a silica matrix has been studied using diffuse reflection spectroscopy. The dependence shows a non monotonic jump-like behaviour indicating a low-temperature size-dependent melting of silver nanoparticles. The melting point decreases with the decrease of the nanoparticle size. The hysteresis in the temperature dependence of the surface plasmon bandwidth has been observed, indicating the first-order phase transition. PMID- 20009175 TI - Au/ITO dual-layer-coated optical fiber probe for multifunctional scanning tunneling microscopy. AB - We have fabricated an Au/ITO (indium-tin-oxide) dual-layer optical fiber probe with functions for high-quality scanning tunneling microscope (STM) imaging and near-field excitation and detection of STM-induced electroluminescence (STML). The inner ITO layer is kept approximately 150 nm in thickness to ensure the conductivity of the fiber probe. The outer Au layer is kept approximately 13 nm in thickness to obtain a better combination of high electron injection efficiency and optical transparency. High-quality STM images, and STML corresponding to maximum quantum efficiencies of 7 x 10(-4) and 1 x 10(-4) photons/electron are obtained on Au(111)/mica and p-GaAs(110) surfaces at room temperature (RT), respectively. The outer Au layer on the Au/ITO-coated fiber probe shows an effective role in improving the quality of STM imaging, the tip apex geometry and durability of the fiber probe, the stability and reproducibility of STML and enhancing the intensity of plasmon-mediated STML from the Au(111) surface but it shows no effect in enhancing the intensity of STML from the GaAs(110) surface. PMID- 20009177 TI - Control of amorphous silica nanowire growth by oxygen content of Si-rich oxide. AB - Ni-coated Si-rich oxide (SRO, SiO(x)) on a p-type Si wafer has been annealed with Si powder to grow silica nanowires (NWs), which have a composition of stoichiometric SiO(2), irrespective of x. The diameters of the NWs are well controlled from 82 to 23 nm by increasing x from 0.4 to 1.2 and they have a uniform distribution at a fixed x. These results suggest that the oxygen content (x) plays a crucial role in determining the diameter of the NWs at the early stage of the NW formation. The growth behaviors of the NWs are explained well based on a modified vapor-liquid-solid mechanism. PMID- 20009176 TI - The electrical conductivity of hydrogenated nanocrystalline silicon investigated at the nanoscale. AB - Hydrogenated nanocrystalline silicon (nc-Si:H) is a multiphase, heterogeneous material, composed of Si nanocrystals embedded in an amorphous matrix. It has been intensively studied in the last few years due to its great promise for photovoltaic and optoelectronics applications. The present paper aims to study the current transport mechanisms in nc-Si:H by mapping the local conductivity at the nanoscale. The role of B doping in nc-Si:H is also investigated. Conductivity maps are obtained by atomic force microscopy using a conductive tip. Differences and similarities between intrinsic and doped nc-Si:H conductivity maps were observed and these are also explained on the basis of recently published computational studies. PMID- 20009178 TI - A highly sensitive and selective fluorescent Cu2+ sensor synthesized with silica nanoparticles. AB - A novel fluorescent nanosensor for the determination of Cu(2+) was synthesized with N-(quinoline-8-yl)-2-(3-triethoxysilyl-propylamino)-acetamide (QlOEt) grafted onto the surface of silica nanoparticles (SiNPs) using the reverse microemulsion method. Spherical SiNPs were used as substrate and QlOEt was used simultaneously as the binding and readout system for Cu(2+). This sensor has been realized as a highly sensitive and selective technique for the detection and quantification of trace amounts of Cu(2+). The probe exhibits a dynamic response range for Cu(2+) from 2.0 x 10(-6) to 2.0 x 10(-5) M, with a detection limit of 3.8 x 10(-7) M. Other alkali, alkaline earth, and transitional metal ions including Li(+), K(+), Mg(2+), Ca(2+), Sr(2+), Mn(2+), Zn(2+), Mo(6+), Pb(2+), Ag(+) had no significant interference on Cu(2+) determination. Poisonous and flammable reagents are avoided during the synthesis of this nanosensor. Therefore the strategy explored in this work can be extended to the synthesis of other chemo- and biosensors for direct detection of specific targets in an intracellular environment. PMID- 20009179 TI - Antioxidant activity and haemolysis prevention efficiency of polyaniline nanofibers. AB - Polyaniline (PAni) nanofibers have been synthesized by interfacial polymerization using hydrochloric acid (HCl) and camphor sulfonic acid (CSA) as dopants. The powder x-ray diffraction pattern of bulk polyaniline reveals ES I structure and has been indexed in a pseudo-orthorhombic lattice. The broadening of (110) reflection in the nanofiber samples has been analysed in terms of domain length and strain using a convolution method employing a Voigt function. The increase in d spacing for the (110) reflection in HCl-doped PAni nanofibers have been assigned to the change in structural conformation due to the increase in the tilt angle of the polymer chain, which is also evident from microRaman spectra. UV-vis spectra of the PAni nanofibers exhibit a remarkable blueshift in the absorption bands attributed to pi-pi* and pi-polaron band transitions indicating a reduction in particle size, which is also observed in TEM micrographs. The antioxidant activity of the polyaniline nanofiber samples has been investigated using 1,1 diphenyl-2-picrylhydrazyl (DPPH) scavenging assay by employing UV-visible spectroscopy. It has also been observed that polyaniline nanofibers are able to protect the haemolysis of red blood cells (RBCs) from cytotoxic agents, namely H(2)O(2). The observed enhancement in the antioxidant and haemolysis prevention activity of the PAni nanofibers as compared to bulk has been attributed to the reduction in particle size and changes in structural conformation, as evident from TEM, XRD and microRaman spectroscopy. PMID- 20009180 TI - Computational dosimetry in embryos exposed to electromagnetic plane waves over the frequency range of 10 MHz-1.5 GHz. AB - This paper presents calculated specific absorption rate (SAR) dosimetry in 4 and 8 week Japanese pregnant-woman models exposed to plane waves over the frequency range of 10 MHz-1.5 GHz. Two types of 2 mm spatial-resolution pregnant-woman models comprised a woman model, which is similar to the average-sized Japanese adult female in height and weight, with a cubic (4 week) embryo or spheroidal (8 week) one. The averaged SAR in the embryos exposed to vertically and horizontally polarized plane waves at four kinds of propagation directions are calculated from 10 MHz to 1.5 GHz. The results indicate that the maximum average SAR in the embryos exposed to plane waves is lower than 0.08 W kg(-1) when the incident power density is at the reference level of ICNIRP guideline for general public environment. PMID- 20009181 TI - FASH and MASH: female and male adult human phantoms based on polygon mesh surfaces: II. Dosimetric calculations. AB - Female and male adult human phantoms, called FASH (Female Adult meSH) and MASH (Male Adult meSH), have been developed in the first part of this study using 3D animation software and anatomical atlases to replace the image-based FAX06 and the MAX06 voxel phantoms. 3D modelling methods allow for phantom development independent from medical images of patients, volunteers or cadavers. The second part of this study investigates the dosimetric implications for organ and tissue equivalent doses due to the anatomical differences between the new and the old phantoms. These differences are mainly caused by the supine position of human bodies during scanning in order to acquire digital images for voxel phantom development. Compared to an upright standing person, in image-based voxel phantoms organs are often coronally shifted towards the head and sometimes the sagittal diameter of the trunk is reduced by a gravitational change of the fat distribution. In addition, volumes of adipose and muscle tissue shielding internal organs are sometimes too small, because adaptation of organ volumes to ICRP-based organ masses often occurs at the expense of general soft tissues, such as adipose, muscle or unspecified soft tissue. These effects have dosimetric consequences, especially for partial body exposure, such as in x-ray diagnosis, but also for whole body external exposure and for internal exposure. Using the EGSnrc Monte Carlo code, internal and external exposure to photons and electrons has been simulated with both pairs of phantoms. The results show differences between organ and tissue equivalent doses for the upright standing FASH/MASH and the image-based supine FAX06/MAX06 phantoms of up to 80% for external exposure and up to 100% for internal exposure. Similar differences were found for external exposure between FASH/MASH and REGINA/REX, the reference voxel phantoms of the International Commission on Radiological Protection. Comparison of effective doses for external photon exposure showed good agreement between FASH/MASH and REGINA/REX, but large differences between FASH/MASH and the mesh-based RPI_AM and the RPI_AF phantoms, developed at the Rensselaer Polytechnic Institute (RPI). PMID- 20009182 TI - The accuracy of pharmacokinetic parameter measurement in DCE-MRI of the breast at 3 T. AB - The purpose of this work is to quantify the accuracy of pharmacokinetic parameter measurement in DCE-MRI of breast cancer at 3 T in relation to three sources of error. Individually, T1 measurement error, temporal resolution and transmitted RF field inhomogeneity are considered. Dynamic contrast enhancement curves were simulated using standard acquisition parameters of a DCE-MRI protocol. Errors on pre-contrast T1 due to incorrect RF spoiling were considered. Flip angle errors were measured and introduced into the fitting routine, and temporal resolution was also varied. The error in fitted pharmacokinetic parameters, K(trans) and v(e), was calculated. Flip angles were found to be reduced by up to 55% of the expected value. The resultant errors in our range of K(trans) and v(e) were found to be up to 66% and 74%, respectively. Incorrect T1 estimation results in K(trans) and v(e) errors up to 531% and 233%, respectively. When the temporal resolution is reduced from 10 to 70 s K(trans) drops by up to 48%, while v(e) shows negligible variation. In combination, uncertainties in tissue T1 map and applied flip angle were shown to contribute to errors of up to 88% in K(trans) and 73% in v(e). These results demonstrate the importance of high temporal resolution, accurate T1 measurement and good B1 homogeneity. PMID- 20009183 TI - FASH and MASH: female and male adult human phantoms based on polygon mesh surfaces: I. Development of the anatomy. AB - Among computational models, voxel phantoms based on computer tomographic (CT), nuclear magnetic resonance (NMR) or colour photographic images of patients, volunteers or cadavers have become popular in recent years. Although being true to nature representations of scanned individuals, voxel phantoms have limitations, especially when walled organs have to be segmented or when volumes of organs or body tissues, like adipose, have to be changed. Additionally, the scanning of patients or volunteers is usually made in supine position, which causes a shift of internal organs towards the ribcage, a compression of the lungs and a reduction of the sagittal diameter especially in the abdominal region compared to the regular anatomy of a person in the upright position, which in turn can influence organ and tissue absorbed or equivalent dose estimates. This study applies tools developed recently in the areas of computer graphics and animated films to the creation and modelling of 3D human organs, tissues, skeletons and bodies based on polygon mesh surfaces. Female and male adult human phantoms, called FASH (Female Adult meSH) and MASH (Male Adult meSH), have been designed using software, such as MakeHuman, Blender, Binvox and ImageJ, based on anatomical atlases, observing at the same time organ masses recommended by the International Commission on Radiological Protection for the male and female reference adult in report no 89. 113 organs, bones and tissues have been modelled in the FASH and the MASH phantoms representing locations for adults in standing posture. Most organ and tissue masses of the voxelized versions agree with corresponding data from ICRP89 within a margin of 2.6%. Comparison with the mesh based male RPI_AM and female RPI_AF phantoms shows differences with respect to the material used, to the software and concepts applied, and to the anatomies created. PMID- 20009184 TI - A fast and pragmatic approach for scatter correction in flat-detector CT using elliptic modeling and iterative optimization. AB - Scattered radiation is a major source of artifacts in flat detector computed tomography (FDCT) due to the increased irradiated volumes. We propose a fast projection-based algorithm for correction of scatter artifacts. The presented algorithm combines a convolution method to determine the spatial distribution of the scatter intensity distribution with an object-size-dependent scaling of the scatter intensity distributions using a priori information generated by Monte Carlo simulations. A projection-based (PBSE) and an image-based (IBSE) strategy for size estimation of the scanned object are presented. Both strategies provide good correction and comparable results; the faster PBSE strategy is recommended. Even with such a fast and simple algorithm that in the PBSE variant does not rely on reconstructed volumes or scatter measurements, it is possible to provide a reasonable scatter correction even for truncated scans. For both simulations and measurements, scatter artifacts were significantly reduced and the algorithm showed stable behavior in the z-direction. For simulated voxelized head, hip and thorax phantoms, a figure of merit Q of 0.82, 0.76 and 0.77 was reached, respectively (Q = 0 for uncorrected, Q = 1 for ideal). For a water phantom with 15 cm diameter, for example, a cupping reduction from 10.8% down to 2.1% was achieved. The performance of the correction method has limitations in the case of measurements using non-ideal detectors, intensity calibration, etc. An iterative approach to overcome most of these limitations was proposed. This approach is based on root finding of a cupping metric and may be useful for other scatter correction methods as well. By this optimization, cupping of the measured water phantom was further reduced down to 0.9%. The algorithm was evaluated on a commercial system including truncated and non-homogeneous clinically relevant objects. PMID- 20009185 TI - The ACB technique: a biomagentic tool for monitoring gastrointestinal contraction directly from smooth muscle in dogs. AB - The aim of this paper was to verify whether AC biosusceptometry (ACB) is suitable for monitoring gastrointestinal (GI) contraction directly from smooth muscle in dogs, comparing with electrical recordings simultaneously. All experiments were performed in dogs with magnetic markers implanted under the serosa of the right colon and distal stomach, and their movements were recorded by ACB. Monopolar electrodes were implanted close to the magnetic markers and their electric potentials were recorded by electromyography (EMG). The effects of neostigmine, hyoscine butylbromide and meal on gastric and colonic parameters were studied. The ACB signal from the distal stomach was very similar to EMG; in the colonic recordings, however, within the same low-frequency band, ACB and EMG signals were characterized by simultaneity or a widely changeable frequency profile with time. ACB recordings were capable of demonstrating the changes in gastric and colonic motility determined by pharmacological interventions as well as by feeding. Our results reinforce the importance of evaluating the mechanical and electrical components of motility and show a temporal association between them. ACB and EMG are complementary for studying motility, with special emphasis on the colon. ACB offers an accurate method for monitoring in vivo GI motility. PMID- 20009186 TI - Enhancing the estimation of blood pressure using pulse arrival time and two confounding factors. AB - A new method of blood pressure (BP) estimation using multiple regression with pulse arrival time (PAT) and two confounding factors was evaluated in clinical and unconstrained monitoring situations. For the first analysis with clinical data, electrocardiogram (ECG), photoplethysmogram (PPG) and invasive BP signals were obtained by a conventional patient monitoring device during surgery. In the second analysis, ECG, PPG and non-invasive BP were measured using systems developed to obtain data under conditions in which the subject was not constrained. To enhance the performance of BP estimation methods, heart rate (HR) and arterial stiffness were considered as confounding factors in regression analysis. The PAT and HR were easily extracted from ECG and PPG signals. For arterial stiffness, the duration from the maximum derivative point to the maximum of the dicrotic notch in the PPG signal, a parameter called TDB, was employed. In two experiments that normally cause BP variation, the correlation between measured BP and the estimated BP was investigated. Multiple-regression analysis with the two confounding factors improved correlation coefficients for diastolic blood pressure and systolic blood pressure to acceptable confidence levels, compared to existing methods that consider PAT only. In addition, reproducibility for the proposed method was determined using constructed test sets. Our results demonstrate that non-invasive, non-intrusive BP estimation can be obtained using methods that can be applied in both clinical and daily healthcare situations. PMID- 20009187 TI - Electrical impedance spectroscopy and diagnosis of tendinitis. AB - There have been a number of studies that investigate the usefulness of bioelectric signals in diagnoses and treatment in the medical field. Tendinitis is a musculoskeletal disorder with a very high rate of occurrence. This study attempts to examine whether electrical impedance spectroscopy (EIS) can detect pathological changes in a tendon and find the exact location of the lesion. Experimental tendinitis was induced by injecting collagenase into one side of the patellar tendons in rabbits, while the other side was used as the control. After measuring the impedance in the tendinitis and intact tendon tissue, the dissipation factor was computed. The real component of impedance and the dissipation factor turned out to be lower in tendinitis than in intact tissues. Moreover, the tendinitis dissipation factor spectrum showed a clear difference from that of the intact tendon, indicating its usefulness as a tool for detecting the location of the lesion. Pathologic findings from the tissues that were obtained after measuring the impedance confirmed the presence of characteristics of tendinitis. In conclusion, EIS is a useful method for diagnosing tendinitis and detecting the lesion location in invasive treatment. PMID- 20009188 TI - Pilot study of temporary controllable gastric pseudobezoars for dynamic non invasive gastric volume reduction. AB - Invasive surgical procedures for gastric volume reduction or bypass have been considered the most effective approach to sustainable long-term weight reduction. However, non-invasive techniques for dynamic volume reduction from inside the stomach are lacking. The aim of this study was to propose temporary, permeable, controllable pseudobezoars for non-invasive, long-term sustainable gastric volume reduction and to test them in pilot human studies. Permeable sac-like carriers made from biocompatible and biodegradable material were filled with expandable superabsorbent fiber and polymer granules. The implements were designed to prevent the expulsion of the pseudobezoars through the pylorus for a controlled time period. The pseudobezoars were administered transorally to two human patients (2M, 78.9 kg/174 cm, girth 88.1 cm, and 89.7 kg/175, girth 95.2 cm). Body weight dynamics, girth, level of satiety, stools, bowel regularity and notable side effects were monitored in three distinct 1 month periods: baseline, therapy and washout. Sonographic verification of the presence of pseudobezoars in the stomachs of both subjects was performed at the end of the therapy month and was repeated at the end of the washout period to examine the clearance of the implements. During the therapy month, both individuals exhibited significant weight and girth reduction (p < 0.05), and substantially increased satiety levels. The patients retained their bowel regularity and did not report any notable side effects. The temporary pseudobezoars were clearly noticeable sonographically in both patients at the end of the therapy month and cleared after its discontinuation. Controllable temporary pseudobezoars were designed and tested in pilot studies. PMID- 20009189 TI - Molecular electroporation and the transduction of oligoarginines. AB - Certain short polycations, such as TAT and polyarginine, rapidly pass through the plasma membranes of mammalian cells by an unknown mechanism called transduction as well as by endocytosis and macropinocytosis. These cell-penetrating peptides (CPPs) promise to be medically useful when fused to biologically active peptides. I offer a simple model in which one or more CPPs and the phosphatidylserines of the inner leaflet form a kind of capacitor with a voltage in excess of about 200 mV, high enough to create a molecular electropore. The model is consistent with an empirical upper limit on the cargo peptide of 40-60 amino acids and with experimental data on how the transduction of a polyarginine-fluorophore into mouse C(2)C(12) myoblasts depends on the number of arginines in the CPP and on the CPP concentration. The model makes three testable predictions. PMID- 20009190 TI - Thermostability in rubredoxin and its relationship to mechanical rigidity. AB - The source of increased stability in proteins from organisms that thrive in extreme thermal environments is not well understood. Previous experimental and theoretical studies have suggested many different features possibly responsible for such thermostability. Many of these thermostabilizing mechanisms can be accounted for in terms of structural rigidity. Thus a plausible hypothesis accounting for this remarkable stability in thermophilic enzymes states that these enzymes have enhanced conformational rigidity at temperatures below their native, functioning temperature. Experimental evidence exists to both support and contradict this supposition. We computationally investigate the relationship between thermostability and rigidity using rubredoxin as a case study. The mechanical rigidity is calculated using atomic models of homologous rubredoxin structures from the hyperthermophile Pyrococcus furiosus and mesophile Clostridium pasteurianum using the FIRST software. A global increase in structural rigidity (equivalently a decrease in flexibility) corresponds to an increase in thermostability. Locally, rigidity differences (between mesophilic and thermophilic structures) agree with differences in protection factors. PMID- 20009191 TI - A statistical approach to the estimation of mechanical unfolding parameters from the unfolding patterns of protein heteropolymers. AB - A statistical calculation is described with which the saw-tooth-like unfolding patterns of concatenated heteropolymeric proteins can be used to estimate the forced unfolding parameters of a previously uncharacterized protein. The chance of observing the various sequences of unfolding events, such as ABAABBB or BBAAABB etc, for two proteins of types A and B is calculated using proteins with various ratios of A and B and at different values of effective unfolding rate constants. If the experimental rate constant for forced unfolding, k(0), and distance to the transition state x(u) are known for one protein, then the calculation allows an estimation of values for the other. The predictions are compared with Monte Carlo simulations and experimental data. PMID- 20009192 TI - Co-registered optical coherence tomography and fluorescence molecular imaging for simultaneous morphological and molecular imaging. AB - Optical coherence tomography (OCT) provides high-resolution, cross-sectional imaging of tissue microstructure in situ and in real time, while fluorescence molecular imaging (FMI) enables the visualization of basic molecular processes. There is a great deal of interest in combining these two modalities so that the tissue's structural and molecular information can be obtained simultaneously. This could greatly benefit biomedical applications such as detecting early diseases and monitoring therapeutic interventions. In this research, an optical system that combines OCT and FMI was developed. The system demonstrated that it could co-register en face OCT and FMI images with a 2.4 x 2.4 mm(2) field-of view. The transverse resolutions of OCT and FMI of the system are both approximately 10 microm. Capillary tubes filled with fluorescent dye Cy 5.5 in different concentrations under a scattering medium are used as the phantom. En face OCT images of the phantoms were obtained and successfully co-registered with FMI images that were acquired simultaneously. A linear relationship between FMI intensity and dye concentration was observed. The relationship between FMI intensity and target fluorescence tube depth measured by OCT images was also observed and compared with theoretical modeling. This relationship could help in correcting reconstructed dye concentration. Imaging of colon polyps of the APC(min) mouse model is presented as an example of biological applications of this co-registered OCT/FMI system. PMID- 20009193 TI - Studies of the interactions of an MRI system with the shielding in a combined PET/MRI scanner. AB - A positron emission tomography (PET) system or 'insert' has been constructed for placement and operation in the bore of a small animal magnetic resonance imaging (MRI) scanner to allow simultaneous MR and PET imaging. The insert contains electronics, components with a variety of magnetic properties and large continuous sheets of metal--all characteristics of an object that should, by conventional wisdom, never be placed in the bore of an MR scanner, especially near the imaging volume. There are a variety of ways the two systems might be expected to interact that could negatively impact the performance of either or both. In this article, the interaction mechanisms, particularly the impact of the PET insert and shielding on MR imaging, are defined and explored. Additionally, some of the difficulties in quantifying errors introduced into the MR images as a result of the presence of the PET components are demonstrated. Several different approaches are used to characterize image artifacts and determine optimal placement of the shielding. Data are also presented that suggest ways the shielding could be modified to reduce errors and enable placement closer to the isocenter of the magnet. PMID- 20009194 TI - A method for partial volume correction of PET-imaged tumor heterogeneity using expectation maximization with a spatially varying point spread function. AB - Tumor heterogeneities observed in positron emission tomography (PET) imaging are frequently compromised by partial volume effects which may affect treatment prognosis, assessment or future implementations such as biologically optimized treatment planning (dose painting). This paper presents a method for partial volume correction of PET-imaged heterogeneous tumors. A point source was scanned on a GE Discovery LS at positions of increasing radii from the scanner's center to obtain the spatially varying point spread function (PSF). PSF images were fit in three dimensions to Gaussian distributions using least squares optimization. Continuous expressions were devised for each Gaussian width as a function of radial distance, allowing for generation of the system PSF at any position in space. A spatially varying partial volume correction (SV-PVC) technique was developed using expectation maximization (EM) and a stopping criterion based on the method's correction matrix generated for each iteration. The SV-PVC was validated using a standard tumor phantom and a tumor heterogeneity phantom and was applied to a heterogeneous patient tumor. SV-PVC results were compared to results obtained from spatially invariant partial volume correction (SINV-PVC), which used directionally uniform three-dimensional kernels. SV-PVC of the standard tumor phantom increased the maximum observed sphere activity by 55 and 40% for 10 and 13 mm diameter spheres, respectively. Tumor heterogeneity phantom results demonstrated that as net changes in the EM correction matrix decreased below 35%, further iterations improved overall quantitative accuracy by less than 1%. SV-PVC of clinically observed tumors frequently exhibited changes of +/-30% in regions of heterogeneity. The SV-PVC method implemented spatially varying kernel widths and automatically determined the number of iterations for optimal restoration, parameters which are arbitrarily chosen in SINV-PVC. Comparing SV PVC to SINV-PVC demonstrated that similar results could be reached using both methods, but large differences result for the arbitrary selection of SINV-PVC parameters. The presented SV-PVC method was performed without user intervention, requiring only a tumor mask as input. Research involving PET-imaged tumor heterogeneity should include correcting for partial volume effects to improve the quantitative accuracy of results. PMID- 20009195 TI - Practical aspects of MR imaging in the presence of conductive guide wires. AB - Various aspects of RF-induced heating of guide wires during their MRI guidance have been investigated in the past. However, the previous works focused on inducing tip heating in either fully immersed or tip-immersed (and otherwise free) wires of impractical lengths in small phantoms. This study simulates real clinical conditions using a product guide wire and a same-length conductive wire partially inserted into a torso-size phantom filled with saline solution. The purpose was to identify potential safety concerns relevant to real clinical applications, as opposed to identifying the worst-case heating scenario. Significant heating occurred at the insertion point, independent of tip heating, with a strong correlation with excitation frequency-dependent imaging parameters. Heat transfer through the wire was also demonstrated to be a safety concern. From these experiments, we have been able to demonstrate additional impacting factors that increase the complexity of safety considerations for the use of conductive guide wires during MR imaging. Safety under a particular set of conditions does not imply safety in all possible conditions that can be encountered during real MRI-guided interventions. PMID- 20009196 TI - Four-dimensional deformable image registration using trajectory modeling. AB - A four-dimensional deformable image registration (4D DIR) algorithm, referred to as 4D local trajectory modeling (4DLTM), is presented and applied to thoracic 4D computed tomography (4DCT) image sets. The theoretical framework on which this algorithm is built exploits the incremental continuity present in 4DCT component images to calculate a dense set of parameterized voxel trajectories through space as functions of time. The spatial accuracy of the 4DLTM algorithm is compared with an alternative registration approach in which component phase to phase (CPP) DIR is utilized to determine the full displacement between maximum inhale and exhale images. A publically available DIR reference database (http://www.dir lab.com) is utilized for the spatial accuracy assessment. The database consists of ten 4DCT image sets and corresponding manually identified landmark points between the maximum phases. A subset of points are propagated through the expiratory 4DCT component images. Cubic polynomials were found to provide sufficient flexibility and spatial accuracy for describing the point trajectories through the expiratory phases. The resulting average spatial error between the maximum phases was 1.25 mm for the 4DLTM and 1.44 mm for the CPP. The 4DLTM method captures the long-range motion between 4DCT extremes with high spatial accuracy. PMID- 20009197 TI - Implementation and evaluation of various demons deformable image registration algorithms on a GPU. AB - Online adaptive radiation therapy (ART) promises the ability to deliver an optimal treatment in response to daily patient anatomic variation. A major technical barrier for the clinical implementation of online ART is the requirement of rapid image segmentation. Deformable image registration (DIR) has been used as an automated segmentation method to transfer tumor/organ contours from the planning image to daily images. However, the current computational time of DIR is insufficient for online ART. In this work, this issue is addressed by using computer graphics processing units (GPUs). A gray-scale-based DIR algorithm called demons and five of its variants were implemented on GPUs using the compute unified device architecture (CUDA) programming environment. The spatial accuracy of these algorithms was evaluated over five sets of pulmonary 4D CT images with an average size of 256 x 256 x 100 and more than 1100 expert-determined landmark point pairs each. For all the testing scenarios presented in this paper, the GPU based DIR computation required around 7 to 11 s to yield an average 3D error ranging from 1.5 to 1.8 mm. It is interesting to find out that the original passive force demons algorithms outperform subsequently proposed variants based on the combination of accuracy, efficiency and ease of implementation. PMID- 20009198 TI - Fluid infusions from catheters into elastic tissue: I. Azimuthally symmetric backflow in homogeneous media. AB - Directly injecting therapeutics into brain tissue has been investigated both experimentally and theoretically. Paul Morrison and others from the National Institutes of Health pointed out the importance of backflow along and outside a catheter inserted into the tissue, once steady state conditions have been reached. Here we investigate and extend their model. We begin with a reformulation of their results and demonstrate an exact solution that exhibits the scaling behavior of the model where the surrounding tissue medium is homogeneous and isotropic. We report on experimental tests of our predictions in agarose gels. We describe the limitations of the assumptions used and the utility of our reformulation. Extensions of the model, including improvements on some of its crude assumptions and generalizations to inhomogeneous media, will be submitted separately. PMID- 20009199 TI - Complexity and accuracy of image registration methods in SPECT-guided radiation therapy. AB - The use of functional imaging in radiotherapy treatment (RT) planning requires accurate co-registration of functional imaging scans to CT scans. We evaluated six methods of image registration for use in SPECT-guided radiotherapy treatment planning. Methods varied in complexity from 3D affine transform based on control points to diffeomorphic demons and level set non-rigid registration. Ten lung cancer patients underwent perfusion SPECT-scans prior to their radiotherapy. CT images from a hybrid SPECT/CT scanner were registered to a planning CT, and then the same transformation was applied to the SPECT images. According to registration evaluation measures computed based on the intensity difference between the registered CT images or based on target registration error, non-rigid registrations provided a higher degree of accuracy than rigid methods. However, due to the irregularities in some of the obtained deformation fields, warping the SPECT using these fields may result in unacceptable changes to the SPECT intensity distribution that would preclude use in RT planning. Moreover, the differences between intensity histograms in the original and registered SPECT image sets were the largest for diffeomorphic demons and level set methods. In conclusion, the use of intensity-based validation measures alone is not sufficient for SPECT/CT registration for RTTP. It was also found that the proper evaluation of image registration requires the use of several accuracy metrics. PMID- 20009200 TI - Automated detection of spinal centrelines, vertebral bodies and intervertebral discs in CT and MR images of lumbar spine. AB - We propose a completely automated algorithm for the detection of the spinal centreline and the centres of vertebral bodies and intervertebral discs in images acquired by computed tomography (CT) and magnetic resonance (MR) imaging. The developed methods are based on the analysis of the geometry of spinal structures and the characteristics of CT and MR images and were evaluated on 29 CT and 13 MR images of lumbar spine. The overall mean distance between the obtained and the ground truth spinal centrelines and centres of vertebral bodies and intervertebral discs were 1.8 +/- 1.1 mm and 2.8 +/- 1.9 mm, respectively, and no considerable differences were detected among the results for CT, T(1)-weighted MR and T(2)-weighted MR images. The knowledge of the location of the spinal centreline and the centres of vertebral bodies and intervertebral discs is valuable for the analysis of the spine. The proposed method may therefore be used to initialize the techniques for labelling and segmentation of vertebrae. PMID- 20009201 TI - Co(OH)3 nanobelts: synthesis, characterization and shape-preserved transformation to pseudo-single-crystalline Co3O4 nanobelts. AB - In this paper, we demonstrated a facile hydrothermal route leading to the generation of a new cobalt hydroxide, Co(OH)(3), nanobelt. This new product of Co(OH)(3) nanobelts was well characterized and identified by SEM, TEM, XRD, EDX, XPS, EXAFS, Raman, TGA and SQUID. Additionally, through systematic investigations on morphological evolution, it was found that the size and shape of Co(OH)(3) nanobelts were adjustable in large scale, e.g. from 50 to 1.5 microm in length and 5 microm to 20 nm in width, by means of fine experimental parameter control. Furthermore, the unique pseudo-single-crystalline Co(3)O(4) nanobelts were produced from Co(OH)(3) precursors via heating treatment. This is the first synthesis of Co(OH)(3) with tunable shapes and sizes, which may find important applications as gas sensors, catalysts, and electrode materials, owing to the specific large surface-to-volume ratio and other unique properties endowed by typical 1D nanobelts. PMID- 20009202 TI - Nanoimprinting using self-assembled ceramic nanoislands. AB - We have combined self-assembled ceramic nanoislands with nanoimprinting to demonstrate a novel, simple, low-cost method for polymer surface patterning. The nanoislands are easy to make and inexpensive, and can produce different distinct island morphologies. With a similar stiffness to steel, the nanoislands have superior durability to silicon, glass, polydimethylsiloxane (PDMS), and other common nanoimprinting materials. The nanoislands are stable up to 1000 degrees C and resist acids, bases, and solvents. We have demonstrated nanoimprinting with PDMS, ethyleneglycol dimethacrylate, and polystyrene polymers. The combination of desirable properties, ease of making, and low cost suggests a useful nanopatterning platform for a wide array of research fields. PMID- 20009203 TI - Engineering graphene by oxidation: a first-principles study. AB - Graphene epoxide, with oxygen atoms lining up on pristine graphene sheets, is investigated theoretically here using first-principles calculations. Two distinct phases-metastable clamped and stable unzipped structures-are observed consistent with experimental observations. In the clamped structure, oxygen atoms form a regular lattice on the graphene sheet. In the unzipped phase, an epoxy group breaks the lower sp(2) bond and modifies the mechanical and electronic properties of graphene remarkably. The foldable epoxy ring structure reduces its Young's modulus by 42.4%, while leaving the tensile strength almost unchanged. The perturbation of epoxidation on the band structures depends on the density and symmetry of oxidation. These results pave the way for oxidation-based engineering of graphene-related materials. PMID- 20009204 TI - Silicon adatom switching and manipulation on Si(111)-7 x 7. AB - We report on a multiple-state switching behavior in the tip height or tunneling current of scanning tunneling microscopy on the Si(111)-7 x 7 surface. This switching is caused by displacement of silicon adatoms under the influence of energetic tunneling electrons. When the tip is fixed over a center adatom, five well-defined levels appear in the measured tip height and tunneling current. These levels are attributed to different electronic structures, depending on the configuration of the center adatoms in the unit cell. We also demonstrate manipulations of the center adatoms by controlling the sample bias. PMID- 20009205 TI - Optimization of multi-walled carbon nanotube-metal contacts by electrical stressing. AB - We present experimental data on the contact resistances of three different metal probes, tungsten, palladium and indium, with chemical vapour deposited (CVD) multi-wall carbon nanotubes (MWCNTs). We demonstrate that there is an irreversible modification of the contacts following electrical stressing whereby the circuit resistance converges towards its optimal value prior to current induced tube failure. Once the probe-MWCNT contact is broken, subsequent recontact experiments reveal that the circuit resistance returns to its initial high level, demonstrating that the modification occurs at the probe contact location and not elsewhere in the circuit. Contact studies with the different metals reveal that Pd metal provides the lowest resistance contact to the MWCNT in our sample. PMID- 20009206 TI - Ultrafast direct imprinting of nanostructures in metals by pulsed laser melting. AB - We report a method of one-step direct patterning of metallic nanostructures. In the method, termed laser assisted direct imprinting (LADI), the surface of a metal film on a substrate is melted by a single excimer laser pulse and subsequently imprinted within approximately 100 ns using a transparent quartz mold, while the substrate is kept at a low temperature and in a solid phase. Using LADI, we imprinted gratings with approximately 100 nm linewidth, 100 nm depth, and 200 nm pitch, as well as isolated mesas of approximately 20 microm size, in Al, Au, Cu and Ni thin films. We found that the quartz mold was able to imprint metals even at temperatures higher than its melting point. The technique could be extended to other metals regardless of their ductility and hardness, and would find applications in photonic and plasmonic device production. PMID- 20009207 TI - Decorating multi-walled carbon nanotubes with quantum dots for construction of multi-color fluorescent nanoprobes. AB - Novel multi-color fluorescent nanoprobes were prepared by electrostatically assembling differently sized CdTe quantum dots on polyethylenimine (PEI) functionalized multi-walled carbon nanotubes (MWNTs). The structural and optical properties of the nano-assemblies (MWNTs-PEI-CdTe) were characterized by transmission electron microscopy (TEM), electron diffraction spectra (EDS), Raman spectroscopy, confocal microscopy and photoluminescence spectroscopy (PL), respectively. Electrochemical impedance spectroscopy (EIS) was also applied to investigate the electrostatic assembling among oxidized MWNTs, PEI and CdTe. Furthermore, confocal fluorescence microscopy was used to monitor the nano assemblies' delivery into tumor cells. It was found that the nano-assemblies exhibit efficient intracellular transporting and strong intracellular tracking. These properties would make this luminescent nano-assembly an excellent building block for the construction of intracellular nanoprobes, which could hold great promise for biomedical applications. PMID- 20009208 TI - Carbon nanotube yarns with high tensile strength made by a twisting and shrinking method. AB - We report a simple and continuous spinning method that combines twisting and shrinking processes to produce carbon nanotube yarns. In this method, a yarn freshly spun from a super-aligned carbon nanotube array is first twisted and then passes through a volatile solvent for shrinking. The as-produced yarn consists of densely packed carbon nanotubes, and thus has a tensile strength up to about 1 GPa. The tensile strength depends on the diameter and the twisting angle of the yarn. Different kinds of solvents, such as water, ethanol, and acetone, are used to shrink the twisted yarns, and acetone shows the best shrinking effect. The origin of the solvent shrinking effect is investigated. Our method is favorable for continuous mass production of high strength carbon nanotube yarns with a wide range of diameters, especially ultra-thin yarns. PMID- 20009209 TI - Site-directed delivery of ferritin-encapsulated gold nanoparticles. AB - Newly designed porter proteins, which catch gold nanoparticles and deliver the nanoparticles selectively to a silicon dioxide (SiO(2)) surface under the specific conditions were reported. Recombinant apoferritin subunits, each of which has gold-binding peptide and titanium-binding peptide at the C- and N terminus, respectively, can efficiently encapsulate a gold nanoparticle. The bio conjugate, a nanogold and surrounding mutant protein subunits, had a property which can deliver itself to the SiO(2) surface through the interaction. In theory, our genetically manipulated apoferritin subunits can encapsulate gold nanoparticles of various sizes, which is a promising property for applications involving surface plasmon resonance. PMID- 20009210 TI - The effect of intrinsic instability of cantilever on static mode atomic force spectroscopy. AB - We show that the static force spectroscopy curve taken in an atomic force microscope is significantly modified due to presence of intrinsic cantilever instability which occurs as a result of its movement in a nonlinear force field. This instability acts in tandem with such instabilities as water bridge or molecular bond rupture and makes the static force spectroscopy curve (including 'jump-off-contact') dependent on the step size of data collection. A theoretical model has been proposed to explain the data. We emphasize the necessity of taking care of this fundamental instability of the microcantilever in calculating the adhesive force and also in the interpretation of data taken using an atomic force microscope. PMID- 20009219 TI - Chronic Leptin Supplementation Ameliorates Pathology and Improves Cognitive Performance in a Transgenic Mouse Model of Alzheimer's Disease. AB - We have previously reported anti-amyloidogenic effects of leptin using in vitro and in vivo models and, more recently, demonstrated the ability of leptin to reduce tau phosphorylation in neuronal cells. The present study examined the efficacy of leptin in ameliorating the Alzheimer's disease (AD)-like pathology in 6-month old CRND8 transgenic mice (TgCRND8) following 8 weeks of treatment. Leptin-treated transgenic mice showed significantly reduced levels of amyloid beta (Abeta){1-40} in both brain extracts (52% reduction, p= 0.047) and serum (55% reduction, p= 0.049), as detected by ELISA, and significant reduction in amyloid burden (47% reduction, p=0.041) in the hippocampus, as detected by immunocytochemistry. The decrease in the levels of Abeta in the brain correlated with a decrease in the levels of C99 C-terminal fragments of the amyloid-beta protein precursor, consistent with a role for beta -secretase in mediating the effect of leptin. In addition, leptin-treated TgCRND8 mice had significantly lower levels of phosphorylated tau, as detected by AT8 and anti-tau-Ser{396} antibodies. Importantly, after 4 or 8 weeks of treatment, there was no significant increase in the levels of C-reactive protein, tumor necrosis factor alpha, and cortisol in the plasma of leptin-treated TgCRND8 animals compared to saline-treated controls, indicating no inflammatory reaction. These biochemical and pathological changes were correlated with behavioral improvements, as early as after 4 weeks of treatment, as recorded by a novel object recognition test and particularly the contextual and cued fear conditioning test after 8 weeks of treatment. Leptin-treated TgCRND8 animals significantly outperformed saline treated littermates in these behavioral tests. These findings solidly demonstrate the potential for leptin as a disease modifying therapeutic in transgenic animals of AD, driving optimism for its safety and efficacy in humans. PMID- 20009237 TI - Genetic variation in the G720/G30 gene locus (DAOA) influences the occurrence of psychotic symptoms in patients with Alzheimer's disease. AB - The occurrence of neuropsychiatric symptoms in patients with Alzheimer's disease hampers the clinical management and exacerbates the burden for caregivers. To what extent psychotic symptoms are genetically determined and which are the genes involved has to be established. We tested the hypothesis that the occurrence of delusions and hallucinations in AD is associated with variations in the G72/DAOA gene, which is supposed to play a key role in the glutamate pathway regulated through the NMDA receptors. A panel of single nucleotide polymorphisms were genotyped in a cohort of 185 Alzheimer's disease patients. The analysis demonstrated a nominally significant association (p< 0.05) with one single nucleotide polymorphism (rs2153674). In addition, multivariate regression showed that the rs2153674 genotype accounts for up to 15% of the variance in delusions severity, as assessed by using the Neuropsychiatric Inventory. If the results from the present study will be replicated, the glutamate hypothesis could be invoked to explain the occurrence of psychosis in neurodegenerative disorders. PMID- 20009238 TI - 12th International Conference on Alzheimer's Disease (ICAD), Vienna, Austria. PMID- 20009242 TI - Microbiological effectiveness of locally produced ceramic filters for drinking water treatment in Cambodia. AB - Low-cost options for the treatment of drinking water at the household level are being explored by the Cambodian government and non-governmental organizations (NGOs) working in Cambodia, where many lack access to improved drinking water sources and diarrhoeal diseases are the most prevalent cause of death in children under 5 years of age. The ceramic water purifier (CWP), a locally produced, low cost ceramic filter, is now being implemented by several NGOs, and an estimated 100,000+households in the country now use them for drinking water treatment. Two candidate filters were tested for the reduction of bacterial and viral surrogates for waterborne pathogens using representative Cambodian drinking water sources (rainwater and surface water) spiked with Escherichia coli and bacteriophage MS2. Results indicate that filters were capable of reducing key microbes in the laboratory with mean reductions of E. coli of approximately 99% and mean reduction of bacteriophages of 90-99% over >600 litres throughput. Increased effectiveness was not observed in filters with an AgNO3 amendment. At under US$10 per filter, locally produced ceramic filters may be a promising option for drinking water treatment and safe storage at the household level. PMID- 20009243 TI - Are the defined substrate-based methods adequate to determine the microbiological quality of natural recreational waters? AB - Monitoring the microbiological quality of water used for recreational activities is very important to human public health. Although the sanitary quality of recreational marine waters could be evaluated by standard methods, they are time consuming and need confirmation. For these reasons, faster and more sensitive methods, such as the defined substrate-based technology, have been developed. In the present work, we have compared the standard method of membrane filtration using Tergitol-TTC agar for total coliforms and Escherichia coli, and Slanetz and Bartley agar for enterococci, and the IDEXX defined substrate technology for these faecal pollution indicators to determine the microbiological quality of natural recreational waters. ISO 17994:2004 standard was used to compare these methods. The IDEXX for total coliforms and E. coli, Colilert, showed higher values than those obtained by the standard method. Enterolert test, for the enumeration of enterococci, showed lower values when compared with the standard method. It may be concluded that more studies to evaluate the precision and accuracy of the rapid tests are required in order to apply them for routine monitoring of marine and freshwater recreational bathing areas. The main advantages of these methods are that they are more specific, feasible and simpler than the standard methodology. PMID- 20009244 TI - Natural and technical factors in faecal contamination incidents of drinking water in small distribution networks, France, 2003-2004: a geographical study. AB - This geographical study aimed to show natural or water-processing-related factors of faecal contamination incidents (FCIs) of drinking water in continental France. We defined a FCI as the occurrence of at least 20 colony-forming Escherichia coli or enterococci among all the 100 mL samples collected for regulatory purpose within one day from a given drinking water supply zone (SZ). We explored correlations between the standardized number of FCIs per departement (N_Pols) and various indicators related to weather, land cover, topography, geology and water management for three SZ size sub-classes. In 2003-2004, 2,739 FCIs occurred in SZs supplying fewer than 2,000 people, mainly with simply disinfected groundwater. N_Pols correlates with four covariates: (1) precipitation; (2) the extension of the karst outcrops; (3) the extent of disinfection; and (4) catchment protection. One hundred millimetres of yearly excess in precipitation increases the pollution risk by 28-37%, depending on the sub-class. A 10% extension of the karst areas, a 10% increase of unprotected resources, or of SZs with no disinfection, could entail a higher risk of FCI by about 10%. The correlations are reproducible over the three sub-classes and corroborate expert appraisals. These results encourage the ongoing effort to generalize disinfection and catchment protection. PMID- 20009245 TI - Estimation of Ascaris infection risks in children under 15 from the consumption of wastewater-irrigated carrots. AB - Ascaris lumbricoides, the large human roundworm, infects approximately 1,200 million people, with children under the age of 15 being particularly at risk. Monte Carlo quantitative microbial risk analyses were undertaken to estimate median Ascaris infection risks in children under 15 from eating raw carrots irrigated with wastewater. For a tolerable additional disease burden of 10(-5) DALY (disability-adjusted life year) loss per person per year (pppy), the tolerable Ascaris infection risk is approximately 10(-3) pppy, which can be achieved in hyperendemic areas by a 4-log unit Ascaris reduction. This reduction can be easily achieved by wastewater treatment in a 1-day anaerobic pond and 5 day facultative pond (2 log units) and peeling prior to consumption (2 log units). PMID- 20009246 TI - Estimation of norovirus infection risks to consumers of wastewater-irrigated food crops eaten raw. AB - A quantitative microbial risk analysis-Monte Carlo method was used to estimate norovirus infection risks to consumers of wastewater-irrigated lettuce. Using the same assumptions as used in the 2006 WHO guidelines for the safe use of wastewater in agriculture, a norovirus reduction of 6 log units was required to achieve a norovirus infection risk of approximately 10(-3) per person per year (pppy), but for a lower consumption of lettuce (40-48 g per week vs. 350 g per week) the required reduction was 5 log units. If the tolerable additional disease burden is increased from a DALY (disability-adjusted life year) loss of 10(-6) pppy (the value used in the WHO guidelines) to 10(-5) pppy, the required pathogen reduction is one order of magnitude lower. Reductions of 4-6 log units can be achieved by very simple partial treatment (principally settling to achieve a 1 log unit reduction) supplemented by very reliable post-treatment health protection control measures such as pathogen die-off (1-2 log units), produce washing in cold water (1 log unit) and produce disinfection (3 log units). PMID- 20009247 TI - Comparison of the presence and partitioning behavior of indicator organisms and Salmonella spp. in an urban watershed. AB - The appropriateness of indicator organisms as surrogates for human pathogens in water quality modeling is dependent on similarities in both presence and transport behavior; however, very little data relating indicator and bacterial pathogen transport behavior in receiving waters is available. In this study observations of presence, partitioning behavior (i.e. association with settleable particles) and removal by upland detention basins were used to assess the suitability of six indicator organisms as surrogates for Salmonella spp. bacteria in an urban watershed. The fecal indicator bacteria (fecal coliforms, E. coli and enterococci) were most closely correlated with Salmonella in terms of presence and partitioning behavior (25-35% associated with settleable particles on average); however, further resolution of the variability associated with Salmonella partitioning is required. Higher removal of particle-associated microbes relative to the total microbial concentration by the detention ponds suggests that sedimentation may be an important removal mechanism. However, large fluctuations in pond performance between storm events and occasional net microbial exports in effluents indicate that these best management practices (BMPs), as currently implemented, will be unlikely to achieve water quality objectives. PMID- 20009248 TI - Turbidity and chlorine demand reduction using alum and moringa flocculation before household chlorination in developing countries. AB - Over 1.1 billion people in the world lack access to improved drinking water. Diarrhoeal and other waterborne diseases cause an estimated 1.87 million deaths per year. The Safe Water System (SWS) is a household water treatment intervention that reduces diarrhoeal disease incidence among users in developing countries. Turbid waters pose a particular challenge to implementation of SWS programmes; although research shows that a 3.75 mg l(-1) sodium hypochlorite dose effectively treats turbid waters, users sometimes object to the strong chlorine taste and prefer to drink water that is more aesthetically pleasing. This study investigated the efficacy of two locally available chemical water treatments-alum and Moringa oleifera flocculation-to reduce turbidity and chlorine demand at turbidities of 10, 30, 70, 100 and 300 NTU. Both treatments effectively reduced turbidity (alum flocculation 23.0-91.4%; moringa flocculation 14.2-96.2%). Alum flocculation effectively reduced chlorine demand compared with controls at 30, 70, 100 and 300 NTU (p=0.01-0.06). Moringa flocculation increased chlorine demand to the point where adequate free chlorine residual was not maintained for 24 hours after treatment. Alum pretreatment is recommended in waters>or=30 NTU for optimum water disinfection. Moringa flocculation is not recommended before chlorination. PMID- 20009249 TI - Free-living amoebae, Legionella and Mycobacterium in tap water supplied by a municipal drinking water utility in the USA. AB - Legionella and Mycobacterium can proliferate within free-living amoebae (FLA) where they are protected from disinfectants at concentrations that can kill bacteria but not protozoa. Despite effective treatment of drinking water, microbes can enter water utility distribution systems (DS) and hence the plumbing within building premises. Additionally, biofilm formation may account for the persistence of microbes in the DS. In the present study a domestic water tap in north-central United States (USA) was sampled in March and September 2007 and analysed for FLA, Legionella and Mycobacterium. Identification of organisms was determined by growth on specific culture media, light and electron microscopy, and amplification of DNA probes specific for each organism. In both the spring and fall samples, amoebae, Legionella and Mycobacterium were detected. However, Acanthamoeba was prominent in the spring sample whereas Vahlkampfia and Naegleria were the amoebae detected in the autumn. Bacterial proliferation in laboratory cultures was noticeably enhanced in the presence of amoebae and biofilms rapidly formed in mixed amoebae and bacteria cultures. It is hypothesized that temperature affected the dynamics of FLA species population structure within the DS and that pathogenic bacteria that proliferate within FLA, which are themselves opportunistic pathogens, pose dual public health risks. PMID- 20009250 TI - Photocatalytic inactivation of Cryptosporidium parvum on nanostructured titanium dioxide films. AB - Control of waterborne gastrointestinal parasites represents a major concern to water industries worldwide. In developed countries, pathogens in drinking water supplies are normally removed by sand filtration followed by chemical disinfection. Cryptosporidium spp. are generally resistant to common disinfection techniques and alternative control strategies are being sought. In the current study, the photocatalytic inactivation of C. parvum oocysts was shown to occur in buffer solution (78.4% after 180 min) and surface water (73.7% after 180 min). Viability was assessed by dye exclusion, excystation, direct examination of oocysts and a novel gene expression assay based on lactate dehydrogenase 1 (LDH1) expression levels. Collectively, this confirmed the inactivation of oocysts and scanning electron microscopy (SEM) confirmed cleavage at the suture line of oocyst cell walls, revealing large numbers of empty (ghost) cells after exposure to photocatalytic treatment. PMID- 20009251 TI - Legionella species and serogroups in Malaysian water cooling towers: identification by latex agglutination and PCR-DNA sequencing of isolates. AB - In this study, we investigated the distribution of Legionella species in water cooling towers located in different parts of Malaysia to obtain information that may inform public health policies for the prevention of legionellosis. A total of 20 water samples were collected from 11 cooling towers located in three different states in east, west and south Malaysia. The samples were concentrated by filtration and treated with an acid buffer before plating on to BCYE agar. Legionella viable counts in these samples ranged from 100 to 2,000 CFU ml(-1); 28 isolates from the 24 samples were examined by latex agglutination as well as 16S rRNA and rpoB PCR-DNA sequencing. These isolates were identified as Legionella pneumophila serogroup 1 (35.7%), L. pneumophila serogroup 2-14 (39%), L. pneumophila non-groupable (10.7%), L. busanensis, L. gormanii, L. anisa and L. gresilensis. L. pneumophila was clearly the predominant species at all sampling sites. Repeat sampling from the same cooling tower and testing different colonies from the same water sample showed concurrent colonization by different serogroups and different species of Legionella in some of the cooling towers. PMID- 20009252 TI - Applicability of quantitative PCR for determination of removal efficacy of enteric viruses and Cryptosporidium by water treatment processes. AB - This study aims to assess the applicability of quantitative PCR (qPCR) for removal studies of adenovirus, coxsackievirus, echovirus and Cryptosporidium by water treatment processes. Bench-scale coagulation jar tests were performed using the enteric viruses and Cryptosporidium. Standard methods (conventional cell culture methods for the viruses and an immunofluorescence assay (IFA) for Cryptosporidium) were used to compare to qPCR. A significant correlation between microbial removals determined by qPCR and the standard detection methods and an approximate 1:1 correlation were observed for the challenge microorganisms. The results indicated that qPCR could be a satisfactory alternate for microbial removal studies using a relative quantification approach. PMID- 20009253 TI - Effect of environmental factors on the fecundity, hatchability and survival of snail Lymnaea (Radix) acuminata (Lamarck): vector of fascioliasis. AB - Lymnaea acuminata breeds round the year. The effect of pH, temperature, dissolved oxygen, carbon dioxide, light/dark period and clean/polluted water on the fecundity, hatchability and survival of young snails of L. acuminata were studied. It was observed that these environmental variant abiotic factors caused a significant variation in fecundity, hatchability and survival of young snails. Maximum reproduction of this snail was observed in the months of March to May. A significant positive correlation (p<0.05) between D.O. (3.1-7.7 ppm)/pH (7.01 8.96) of water with fecundity (6.0-196.33/20 snails), hatchability (54.69-96.91%) and survival (61.3-95.86%) of young snails was observed for each month and each interval of 24-72 h. In contrast, a significant negative correlation between dissolved CO2 (4.6-16.6 ppm)/temperature (16-37 degrees C) of water was noted with fecundity, hatchability and survival of snails. Percent hatchability in the eggs in different regimens of water was between 96.91-54.69%. The hatching period was prolonged (2-14 days) in snail exposed to different groups of water compared to the control group (2-9 d). This study conclusively shows that variant abiotic factors in different months of the year can significantly alter the reproductive ability and development process in the snail Lymnaea acuminata. PMID- 20009254 TI - Simultaneous determination of 42 organic chemicals in bottled water by combining C18 extraction disk with GC-MS and LC/MS/MS technique. AB - A method for the determination of 42 hazard residues required by 'Japan Positive List System' in bottled water was described. Hazard compounds in bottled water were extracted with a solid phase extraction step using C18 disks. Determination was carried out by gas chromatography/mass spectrometry (GC/MS) and liquid chromatography-tandem mass spectrometry (LC/MS/MS). The disk extraction has high throughput which is well adapted to isolate and enrich these compounds from large volumes of water. For the water sample spiked at three concentration levels (LOQ, 4 times LOQ and 8 times LOQ), the recoveries of all analytes ranged between 65% and 120% with a relative standard deviation<24% (n=8). PMID- 20009255 TI - Microbial and metal water quality in rain catchments compared with traditional drinking water sources in the East Sepik Province, Papua New Guinea. AB - In Papua New Guinea, a significant portion of morbidity and mortality is attributed to water-borne diseases. To reduce incidence of disease, communities and non-governmental organizations have installed rain catchments to provide drinking water of improved quality. However, little work has been done to determine whether these rain catchments provide drinking water of better quality than traditional drinking water sources, and if morbidity is decreased in villages with rain catchments. The specific aim of this study was to evaluate the quality of water produced by rain catchments in comparison with traditional drinking water sources in rural villages in the East Sepik Province. Fifty-four water sources in 22 villages were evaluated for enterococci and Escherichia coli densities as well as 14 health-relevant metals. In addition, we examined how the prevalence of diarrhoeal illness in villages relates to the type of primary drinking water source. The majority of tested metals were below World Health Organization safety limits. Catchment water sources had lower enterococci and E. coli than other water sources. Individuals in villages using Sepik River water as their primary water source had significantly higher incidence of diarrhoea than those primarily using other water sources (streams, dug wells and catchments). PMID- 20009256 TI - Risks of childhood cancer among Texas watersheds, based on mothers' living locations at the time of birth. AB - Cancer is the most common fatal disease among US children. The fetus has reduced resistance to toxic injury and is especially prone to mutagenic injury because of the high rate of cell division. A fetus can be exposed to environmental toxins through maternal consumption of contaminated water. The objective of this study was to estimate the incidence risk for childhood cancers within each watershed in Texas. The approach modeled risk for 19 cancer histotypes incorporating correlations among the cancer types and spatial correlation. Several watersheds in a very large area known as the Central Great Plains of North Texas were associated with increased risk for astrocytoma. Two watersheds near Houston, Buffalo-San Jacinto and West Galveston Bay, had increased risk for renal cancer and acute lymphoid leukemia, respectively. A watershed in South Texas, the South Laguna Madre, had increased risk for atypical leukemias. The possibility that waterborne toxins cause these childhood cancers should be investigated further. PMID- 20009257 TI - Comparative analysis of the composition of bacterial communities from two constructed wetlands for municipal and swine wastewater treatment. AB - This work provides information about bacterial community structure in natural wastewater treatment systems treating different types of wastewater. The diversity and composition of bacterial communities associated with the rhizosphere of Typha latifolia and Salix atrocinerea were studied and compared among two different natural wastewater treatment systems, using the direct sequencing of the 16S ribosomal RNA codifying genes. Phylogenetic affiliations of the bacteria detected allowed us to define the main groups present in these particular ecosystems. Moreover, bacterial community structure was studied through two diversity indices. Ten identified and five non-identified phyla were found in the samples; the phylum Proteobacteria was the predominant group in the four ecosystems. The results showed a bacterial community dominated by beta proteobacteria and a lower diversity value in the swine wastewater treatment system. The municipal wastewater treatment system presented a high diverse community in both macrophytes (Typha latifolia and Salix atrocinerea), with gamma proteobacteria and alpha-proteobacteria, respectively, as the most abundant groups. PMID- 20009258 TI - One year monitoring of norovirus in a sewage treatment plant in Rio de Janeiro, Brazil. AB - Norovirus (NoV) is one of the most important aetiological agents of acute gastroenteritis both in developed and developing countries. NoV is shed in high concentrations by infected persons and contaminates recreational and drinking water through sewage discharge into the environment. The aim of this study was to determine the prevalence, genotypes and removal ratio of NoV by PCR, seminested PCR and quantitative PCR (qPCR) assays in a sewage treatment plant in Rio de Janeiro city, Brazil, during one year of surveillance. NoV was detected in 7 (15%), 14 (29%) and 28 (58%) samples using PCR, seminested-PCR and qPCR, respectively. The mean removal ratio for the activated sludge process was 0.6 log10 for NoV genogroup I (GI) and 0.32 log10 for NoV genogroup II (GII). The peak NoV concentration was detected in the coldest months, with 53,300 genomic copies/litre. Nucleotide sequencing and phylogenetic analysis revealed that five strains clustered with GI strains and six with GII strains. This study demonstrates that NoV spreads into the environment despite the sewage treatment process and remains a source of waterborne outbreaks of acute gastroenteritis. PMID- 20009259 TI - Modelling latrine diffusion in Benin: towards a community typology of demand for improved sanitation in developing countries. AB - Latrine diffusion patterns across 502 villages in Benin, West Africa, were analysed to explore factors driving initial and increasing levels of household adoption in low-coverage rural areas of sub-Saharan Africa. Variables explaining adoption related to population density, size, infrastructure/services, non agricultural occupations, road and urban proximity, and the nearby latrine adoption rate, capturing differences in the physical and social environment, lifestyles and latrine exposure involved in stimulating status/prestige and well being reasons for latrine adoption. Contagion was most important in explaining adoption initiation. Cluster analysis revealed four distinct village typologies of demand for latrines which provide a framework for tailoring promotional interventions to better match the different sanitation demand characteristics of communities in scaling-up sanitation development and promotion programmes. PMID- 20009260 TI - Social-epidemiological study for evaluation of water supply and sanitation systems of low-income urban community in Dhaka, Bangladesh. AB - This study aims at quantification of health losses, considering social and environmental factors. Morbidity and mortality cases of diarrhoea for children under five years old were used to estimate the disability adjusted life years (DALYs) lost for the target households in low-income communities in Dhaka, Bangladesh. Water supply facilities and sanitation systems, along with hygiene practices and their health outcomes, were studied at community level. Demographic, socio-economic and socio-cultural aspects were also studied to support the research findings and give a better understanding of the local conditions. The four selected communities, Ward 60 (W60), Ward 61 (W61), Ward 62 (W62) and Ward 65 (W65), all had different existing urban services such as water supply, sanitation, garbage management and drainage facilities. All of these services existed in W62, but W60 did not have any of the services; W61 had sanitation and drainage coverage, whereas W65 had only a water supply facility. The results conclusively showed that, compared with the null (absence of services) scenario (W60), the other three scenarios (W61, W62 and W65) showed a substantial decrease of diarrhoea (1.219, 1.284 and 2.052 DALYs/household/year, respectively) reported for children under five years old. Besides urban services, other socio-economic characteristics might also influence the prevalence of diseases. PMID- 20009262 TI - Divergence time estimates of mammals from molecular clocks and fossils: relevance of new fossil finds from India. AB - This paper presents a brief review of recent advances in the classification of mammals at higher levels using fossils and molecular clocks. It also discusses latest fossil discoveries from the Cretaceous - Eocene (66-55 m.y.) rocks of India and their relevance to our current understanding of placental mammal origins and diversifications. PMID- 20009261 TI - The association of drinking water quality and sewage disposal with Helicobacter pylori incidence in infants: the potential role of water-borne transmission. AB - The mode of transmission of Helicobacter pylori, a bacterium causing gastric cancer and peptic ulcer disease, is unknown although waterborne transmission is a likely pathway. This study investigated the hypothesis that access to treated water and a sanitary sewerage system reduces the H. pylori incidence rate, using data from 472 participants in a cohort study that followed children in Juarez, Mexico, and El Paso, Texas, from April 1998, with caretaker interviews and the urea breath test for detecting H. pylori infection at target intervals of six months from birth through 24 months of age. The unadjusted hazard ratio comparing bottled/vending machine water to a municipal water supply was 0.71 (95% confidence interval (CI): 0.50, 1.01) and comparing a municipal sewer connection to a septic tank or cesspool, 0.85 (95% CI: 0.60, 1.20). After adjustment for maternal education and country, the hazard ratios decreased slightly to 0.70 (95% confidence interval: 0.49, 1.00) and 0.77 (95% confidence interval: 0.50, 1.21), respectively. These results provide moderate support for potential waterborne transmission of H. pylori. PMID- 20009263 TI - Growth patterns of fossil vertebrates as deduced from bone microstructure: case studies from India. AB - Bone microstructure is affected by ontogeny, phylogeny, biomechanics and environments. These aspects of life history of an extinct animal, especially its growth patterns, may be assessed as fossil bone generally maintains its histological integrity. Recent studies on the bone histology of fossil vertebrates from India encompass different types of temnospondyls and dicynodonts from different Permian and Triassic horizons. The examined taxa show that they had distinct bone histology and varied growth patterns. The Early Triassic trematosaurids had an overall fast growth, which contrasts with that of the Middle and Late Triassic temnospondyl taxa examined. The dicynodonts on the other hand, were characterized by an overall fast growth with periodic interruptions, variable growth rates dependent on ontogeny and indeterminate growth strategy. A comparative study encompassing several neotherapsid genera including the dicynodonts shows signifi cant evolutionary trends towards determinate growth strategy and reduced developmental plasticity. PMID- 20009264 TI - The origin and early evolution of whales: macroevolution documented on the Indian subcontinent. AB - The origin of whales (order Cetacea) from a four-footed land animal is one of the best understood examples of macroevolutionary change. This evolutionary transition has been substantially elucidated by fossil finds from the Indian subcontinent in the past decade and a half. Here, we review the first steps of whale evolution, i.e. the transition from a land mammal to obligate marine predators, documented by the Eocene cetacean families of the Indian subcontinent: Pakicetidae, Ambulocetidae, Remingtonocetidae, Protocetidae, and Basilosauridae, as well as their artiodactyl sister group, the Raoellidae. We also discuss the influence that the excellent fossil record has on the study of the evolution of organ systems, in particular the locomotor and hearing systems. PMID- 20009265 TI - The out-of-India hypothesis: what do molecules suggest? AB - The remarkable geological and evolutionary history of peninsular India has generated much interest in the patterns and processes that might have shaped the current distributions of its endemic biota. In this regard the "Out-of-India" hypothesis, which proposes that rafting peninsular India carried Gondwanan forms to Asia after the break-up of Gondwana super continent, has gained prominence. Here we have reviewed molecular studies undertaken on a range of taxa of supposedly Gondwanan origin to better understand the Out-of-India scenario. This re-evaluation of published molecular studies indicates that there is mounting evidence supporting Out-of-India scenario for various Asian taxa. Nevertheless, in many studies the evidence is inconclusive due to lack of information on the age of relevant nodes. Studies also indicate that not all Gondwanan forms of peninsular India dispersed out of India. Many of these ancient lineages are confi ned to peninsular India and therefore are relict Gondwanan lineages. Additionally, for some taxa an "Into India" rather than "Out-of-India" scenario better explains their current distribution. To identify the "Out-of-India" component of Asian biota it is imperative that we understand the complex biogeographical history of India. To this end, we propose three oversimplified yet explicit phylogenetic predictions. These predictions can be tested through the use of molecular phylogenetic tools in conjunction with palaeontological and geological data. PMID- 20009266 TI - Tree ring imprints of long-term changes in climate in western Himalaya, India. AB - Tree-ring analyses from semi-arid to arid regions in western Himalaya show immense potential for developing millennia long climate records. Millennium and longer ring-width chronologies of Himalayan pencil juniper (Juniperus polycarpos), Himalayan pencil cedar (Cedrus deodara) and Chilgoza pine (Pinus gerardiana) have been developed from different sites in western Himalaya. Studies conducted so far on various conifer species indicate strong precipitation signatures in ring-width measurement series. The paucity of weather records from stations close to tree-ring sampling sites poses diffi culty in calibrating tree ring data against climate data especially precipitation for its strong spatial variability in mountain regions. However, for the existence of strong coherence in temperature, even in data from distant stations, more robust temperature reconstructions representing regional and hemispheric signatures have been developed. Tree-ring records from the region indicate multi-century warm and cool anomalies consistent with the Medieval Warm Period and Little Ice Age anomalies. Signifi cant relationships noted between mean premonsoon temperature over the western Himalaya and ENSO features endorse utility of climate records from western Himalayan region in understanding long-term climate variability and attribution of anthropogenic impact. PMID- 20009267 TI - Deccan volcanism, the KT mass extinction and dinosaurs. AB - Recent advances in Deccan volcanic studies indicate three volcanic phases with the phase-1 at 67.5 Ma followed by a 2 m.y. period of quiescence. Phase-2 marks the main Deccan volcanic eruptions in Chron 29r near the end of the Maastrichtian and accounts for approximately 80% of the entire 3500 m thick Deccan lava pile. At least four of the world's longest lava flows spanning 1000 km across India and out into the Gulf of Bengal mark phase-2. The final phase-3 was smaller, coincided with the early Danian Chron 29n and also witnessed several of the longest lava flows. The KT boundary and mass extinction was first discovered based on planktic foraminifera from shallow marine intertrappean sediments exposed in Rajahmundry quarries between the longest lava flows of the main volcanic phase-2 and smaller phase-3. At this locality early Danian (zone P1a) planktic foraminiferal assemblages directly overlie the top of phase-2 eruptions and indicate that the masse extinction coincided with the end of this volcanic phase. Planktic foraminiferal assemblages also mark the KT boundary in intertrappean sediments at Jhilmili, Chhindwara, where freshwater to estuarine conditions prevailed during the early Danian and indicate the presence of a marine seaway across India at KT time. Dinosaur bones, nesting sites with complete eggs and abundant eggshells are known from central India surrounding the hypothesized seaway through the Narmada-Tapti rift zone. A Maastrichtian age is generally assigned to these dinosaur remains. Age control may now be improved based on marine microfossils from sequences deposited in the seaway and correlating these strata to nearby terrestrial sequences with dinosaur remains. PMID- 20009269 TI - Deep-sea palaeoceanography of the Maldives islands (ODP hole 716A), equatorial Indian ocean during MIS 12-6. AB - Deep-sea benthic foraminifera, planktic foraminifer Globigerina bulloides and pteropods have been quantitatively analysed in 451 samples from Ocean Drilling Program (ODP) Hole 716A, to understand both surface and deep-sea palaeoceanographic changes in the equatorial Indian Ocean basin during the late Quaternary (approximately 444-151 Kyrs). Benthic foraminifera were analysed from >125 microm size fraction whereas Globigerina bulloides and pteropods were analysed from > 150 microm size fraction. Factor analysis of most dominant benthic foraminiferal species over the studied time span made it possible to identify three biofacies characterizing distinct deep-sea environmental settings at Hole 716A. The environmental interpretation of each species is based on the ecology of recent deep-sea benthic foraminifera. The faunal record indicates fluctuating deep-sea conditions including changes in surface productivity, organic food supply and deep-sea oxygenation linked to changing wind intensities. These changes are pronounced on glacial-interglacial time scales driven by summer monsoon winds. PMID- 20009268 TI - India at the cross-roads of human evolution. AB - The Indian palaeoanthropological record, although patchy at the moment, is improving rapidly with every new find. This broad review attempts to provide an account of (a) the Late Miocene fossil apes and their gradual disappearance due to ecological shift from forest dominated to grassland dominated ecosystem around 9-8 Ma ago, (b) the Pliocene immigration/evolution of possible hominids and associated fauna, (c) the Pleistocene record of fossil hominins, associated fauna and artifacts, and (d) the Holocene time of permanent settlements and the genetic data from various human cultural groups within India. Around 13 Ma ago (late Middle Miocene) Siwalik forests saw the emergence of an orangutan-like primate Sivapithecus. By 8 Ma, this genus disappeared from the Siwalik region as its habitat started shrinking due to increased aridity influenced by global cooling and monsoon intensification. A contemporary and a close relative of Sivapithecus, Gigantopithecus (Indopithecus), the largest ape that ever-lived, made its first appearance at around 9 Ma. Other smaller primates that were pene-contemporaneous with these apes were Pliopithecus (Dendropithecus), Indraloris, Sivaladapis and Palaeotupia. The Late Pliocene and Early Pleistocene witnessed northern hemisphere glaciations, followed by the spread of arid conditions on a global scale, setting the stage for hominids to explore "Savanahastan". With the prominent expansion of grassland environments from East Africa to China and Indonesia in the Pliocene, monkeys and baboons dispersed into the Indian subcontinent from Africa along with other mammals. Though debated, there are several claims of the presence of early hominins in this part of the world during the Late Pliocene, based primarily on the recovery of Palaeolithic tools. Fossils of our own ancestor and one of the first globe-trotters, early Homo erectus, has been documented from the Early Pleistocene of East Africa, Western Asia and Southeast Asia, thus indirectly pointing towards Indian subcontinent as a possible migration corridor between these regions. The only definite pre-Homo sapiens fossil hominin remains come from the Central Narmada Valley and are thought to be of Middle to late Pleistocene age, and the cranium has been shown to be closely linked to archaic Homo sapiens/H. heidelbergensis of Europe. Around approximately 74,000 yrs ago, a super volcanic eruption in Sumatra caused the deposition of Youngest Toba Tephra, that covered large parts of the Indian peninsula. Just around this time anatomically-and-behaviorally modern humans or Homo sapiens possibly arrived into India as evidenced by the so called Middle and Upper Palaeolithic assemblages and associated symbolic evidence. The available genetic data reveals that the gene pool to which modern Indians races belong was extremely diverse and had variable mixed links with both European and Asian populations. PMID- 20009270 TI - The evolution and distribution of life in the Precambrian eon-global perspective and the Indian record. AB - The discovery of Precambrian microfossils in 1954 opened a new vista of investigations in the field of evolution of life. Although the Precambrian encompasses 87% of the earth's history, the pace of organismal evolution was quite slow. The life forms as categorised today in the three principal domains viz. the Bacteria, the Archaea and the Eucarya evolved during this period. In this paper, we review the advancements made in the Precambrian palaeontology and its contribution in understanding the evolution of life forms on earth. These studies have enriched the data base on the Precambrian life. Most of the direct evidence includes fossil prokaryotes, protists, advanced algal fossils, acritarchs, and the indirect evidence is represented by the stromatolites, trace fossils and geochemical fossils signatures. The Precambrian fossils are preserved in the form of compressions, impressions, and permineralized and biomineralized remains. PMID- 20009271 TI - Evidence of late Palaeocene-early Eocene equatorial rain forest refugia in southern Western Ghats, India. AB - Equatorial rain forests that maintain a balance between speciation and extinction are hot-spots for studies of biodiversity. Western Ghats in southern India have gained attention due to high tropical biodiversity and endemism in their southern most area. We attempted to track the affinities of the pollen fl ora of the endemic plants of Western Ghat area within the fossil palynoflora of late Palaeocene-early Eocene (approximately 55-50 Ma) sedimentary deposits of western and northeastern Indian region. The study shows striking similarity of extant pollen with twenty eight most common fossil pollen taxa of the early Palaeogene. Widespread occurrences of coal and lignite deposits during early Palaeogene provide evidence of existence of well diversified rain forest community and swampy vegetation in the coastal low lying areas all along the western and northeastern margins of the Indian subcontinent. Prevalence of excessive humid climate during this period has been seen as a result of equatorial positioning of Indian subcontinent, superimposed by a long term global warming phase (PETM and EECO) during the early Palaeogene. The study presents clear evidence that highly diversifi ed equatorial rain forest vegetation once widespread in the Indian subcontinent during early Palaeogene times, are now restricted in a small area as a refugia in the southernmost part of the Western Ghat area. High precipitation and shorter periods of dry months seem to have provided suitable environment to sustain lineages of ancient tropical vegetation in this area of Western Ghats in spite of dramatic climatic changes subsequent to the post India-Asia collision and during the Quaternary and Recent times. PMID- 20009272 TI - Biomechanical aspects of bone microstructure in vertebrates: potential approach to palaeontological investigations. AB - Biomechanical or biophysical principles can be applied to study biological structures in their modern or fossil form. Bone is an important tissue in paleontological studies as it is a commonly preserved element in most fossil vertebrates, and can often allow its microstructures such as lacuna and canaliculi to be studied in detail. In this context, the principles of Fluid Mechanics and Scaling Laws have been previously applied to enhance the understanding of bone microarchitecture and their implications for the evolution of hydraulic structures to transport fluid. It has been shown that the microstructure of bone has evolved to maintain efficient transport between the nutrient supply and cells, the living components of the tissue. Application of the principle of minimal expenditure of energy to this analysis shows that the path distance comprising five or six lamellar regions represents an effective limit for fluid and solute transport between the nutrient supply and cells; beyond this threshold, hydraulic resistance in the network increases and additional energy expenditure is necessary for further transportation. This suggests an optimization of the size of the bone's building blocks (such as osteon or trabecular thickness) to meet the metabolic demand concomitant to minimal expenditure of energy. This biomechanical aspect of bone microstructure is corroborated from the ratio of osteon to Haversian canal diameters and scaling constants of several mammals considered in this study. This aspect of vertebrate bone microstructure and physiology may provide a basis of understanding of the form and function relationship in both extinct and extant taxa. PMID- 20009273 TI - Palynoflora from Deccan volcano-sedimentary sequence (Cretaceous-Palaeogene transition) of central India: implications for spatio-temporal correlation. AB - The sedimentary beds associated with Deccan Continental Flood Basalt (DCFB) sequences exposed in the volcanic subprovinces of Jabalpur-Mandla-Chhindwara (JMC) regions of Madhya Pradesh and Nand-Dongargaon (N-D) basin and the adjoining areas to the west in Yeotmal-Nanded in Maharashtra were studied for their palynofloral analysis. The sediments were characterized palynologically and changes in the palynoflora are observed at different stratigraphic levels in a number of sections including several new intertrappean localities recorded in recent years. For the purpose of effective correlation of different subprovinces, palynofloras of some of the previously studied intertrappeans are also reviewed. Our studies suggest that before the start of the Deccan volcanic activity, the palynoflora found in the Lameta sediments, was dominated by gymnosperms angiosperm association. The plant canopy consisted mainly of gymnosperms (Conifers and Podocarpaceae) whereas, the understory members were mostly of palms and herbs (Poaceae and Asteraceae). The eruption of Deccan volcanic flows severely affected the existing floral association and proved fatal for the well established plant community. The immediately overlying sediments associated with the earliest volcanic flows are dominated by pteridophytes and angiosperm taxa (Azolla cretacea, Aquilapollenites bengalensis, Ariadnaesporites sp., Gabonisporis vigourouxii and Triporoletes reticulatus). Higher up in the stratigraphic sequence, similar forms continued with simultaneous appearance of new taxa including Scabrastephanocolpites spp. At still higher stratigraphic levels, abundance of fungi especially the mycorrhizal fungi, concurrent with sharp decline in pollen/spore recovery was observed. In the culminating phase (i.e. Palaeocene) of Deccan volcanic history a new palynofloral assemblage of typical Palaeocene taxa (Dandotiaspora dilata, D. pseudoauriculata, D. plicata, Spinizonocolpites echinatus, Matanomadhiasulcites sp., and Lakiapollis ovatus) was encountered. PMID- 20009274 TI - Passive immunity to pandemic H1N1 2009 by swine flu parties. AB - The general population is concerned about the probable devastating effects of pandemic H1N1 2009. Based upon the 1918 Spanish flu pandemic, scientific publications and theories, the idea of swine flu parties to achieve passive immunity against pandemic H1N1 2009 has been proposed. Public health officials have asked the general public not to resort to this method. However, no concrete evidence of the reasoning behind the recommendation has been given. In this paper, we have dynamically modeled the effect of swine flu parties on the immunity achieved and associated mortality for a period of two years. The simulations show that the public should not organize or participate in swine flu parties as they will likely increase swine flu-associated mortality. PMID- 20009275 TI - Chikungunya: an emerging and spreading arthropod-borne viral disease. AB - The virus causing Chikungunya disease was identified over 50 years ago; however, because the disease appeared only in developing countries, little research on it has been done. Research interest in the disease increased after an important epidemiological outbreak occurred in 2005 on the French metropolitan island of La Reunion located in the south-eastern part of the Indian Ocean. In 2007, a smaller outbreak of Chikungunya developed in the north-eastern part of Italy made possible by immigration of a viremic patient from the Indian Ocean area and the enormous population of Aedes albopictus in Italy. Currently, Chikungunya is spreading in Southeast Asian aspects, clinical pictures, diagnosis and treatment of the disease caused by Chikungunya virus. PMID- 20009276 TI - Enteric fever in Mediterranean north Africa. AB - Typhoid fever is endemic in the Mediterranean North African countries (Morocco, Algeria, Tunisia, Libya, and Egypt) with an estimated incidence of 10-100 cases per 100,000 persons. Outbreaks caused by Salmonella enterica serovar Typhi are common and mainly due to the consumption of untreated or sewage-contaminated water. Salmonella enterica Paratyphi B is more commonly involved in nosocomial cases of enteric fever in North Africa than expected and leads to high mortality rates among infants with congenital anomalies. Prevalence among travellers returning from this region is low, with an estimate of less than one per 100,000. Although multidrug resistant strains of Salmonella Typhi and Paratyphi are prevalent in this region, the re-emergence of chloramphenicol- and ampicillin susceptible strains has been observed. In order to better understand the epidemiology of enteric fever in the Mediterranean North African region, population-based studies are needed. These will assist the health authorities in the region in preventing and controlling this important disease. PMID- 20009277 TI - Effect of Ibicella lutea on uropathogenic Proteus mirabilis growth, virulence, and biofilm formation. AB - BACKGROUND: Proteus mirabilis, an important uropathogen that can cause complicated urinary tract infections (UTI), has emerged as a therapeutic problem following mutations that compromise the use of antimicrobial drugs. Due to the serious effects associated with uropathogenic P. mirabilis and the problems related to the use of antibiotics, it is necessary to develop alternative strategies for its control. The objective of this study was to assess the effect of Ibicella lutea extract, a South American indigenous plant, on growth, virulence and biofilm production of uropathogenic P. mirabilis. METHODOLOGY: This study was based on the extract generation and the assessment of its effect on bacterial features related to virulence. These assays involved determination of antibacterial activity, swarming motility, Western blot to assess expression of fimbriae and flagella, biofilms formation, haemagglutination, haemolysis, and electron microscopy. RESULTS AND CONCLUSIONS: I. lutea extract had an effect on bacterial growth rate and bacterial morphology. It also affected P. mirabilis swarming differentiation, hemagglutination and biofilm formation on glass and polystyrene. These findings suggest that I. lutea may have a role as an agent for the control of P. mirabilis UTI. PMID- 20009278 TI - Ventilator-associated pneumonia in a tertiary care hospital in India: incidence and risk factors. AB - BACKGROUND: Knowledge of the incidence of ventilator-associated pneumonia (VAP) and its associated risk factors is imperative for the development and use of more effective preventive measures. METHODOLOGY: We performed a prospective study over a period of 15 months to determine the incidence and the risk factors for development of VAP in critically ill adult patients admitted in different intensive care units (ICUs) of Jawaharlal Institute of Post-graduate Medical Education and Research (JIPMER), a tertiary care hospital in Pondicherry, India. RESULTS: The incidence of VAP was 30.67 and 15.87 per 1,000 ventilator days in the two different ICUs. In our study 58.3% of the cases were late-onset VAP, while 41.7% were early-onset VAP. Univariate analysis indicated that the following were significantly associated with VAP: impaired consciousness, tracheostomy, re-intubation, emergency intubation, and nasogastric tube. Emergency intubation and intravenous sedatives were found to be the specific risk factors for early onset VAP, while tracheostomy and re-intubation were the independent predictors of late-onset VAP by multivariate logistic regression analysis. CONCLUSIONS: Knowledge of these risk factors may be useful in implementing simple and effective preventive measures including non-invasive ventilation, precaution during emergency intubation, minimizing the occurrence of reintubation, avoidance of tracheostomy as far as possible, and minimization of sedation. PMID- 20009279 TI - Indirect patient expenses for antituberculosis treatment in Tijuana, Mexico: is treatment really free? AB - BACKGROUND: One of the main problems faced by the Mexican National Tuberculosis Program is the high rate of patients abandoning treatment. This study aimed to determine the magnitude of unaccounted costs of tuberculosis (TB) treatment in Tijuana, Mexico. METHODOLOGY: Subjects were recruited at 21 health centres. Patients had confirmed active pulmonary TB, had been on treatment for more than 12 weeks, and were aged 18 years and older. The questionnaire provided information about demographics, past and current episodes of TB, and various categories of expenses. RESULTS: The study included 180 patients as follows: 48 had been diagnosed with tuberculosis in the past (26.6%) and had either currently relapsed or failed treatment; 160 (88.8%) were under directly observed therapy (DOT); 131 (72.8%) attended a health centre; and the rest received directly observed treatment at home. The daily cost of transportation to the health centre was MXN $25.88 +/- 3.22 (1 USD = 13 MXN). Thirty-two patients (17.8%) had to buy medication at least once, with a monthly medication expense of MXN $440.5 +/- 40.3. Patients receiving DOT at the health centre reported daily food and beverages expenses, spending MXN $56.5 +/- 10.1. Forty-two patients reported laboratory testing expenses, on average MXN $558.8 +/- 85.8 per month. Eighty patients (42.4%) reported expenses on radiographic/ultrasound studies, on average MXN $562.9 +/- 72.1 per six-month regimen. Conclusions TB diagnosis and treatment posed a significant economic burden on patients in terms of both cost and affordability; clinic-based DOT may contribute disproportionately to the costs incurred by patients. PMID- 20009280 TI - Knowledge and attitudes of tuberculosis management in San Juan de Lurigancho district of Lima, Peru. AB - BACKGROUND: Expansion of the health care workforce in Peru to combat tuberculosis (TB) includes both professional health care providers (HCPs) such as doctors and nurses, and non-professional HCPs such as community health workers (CHWs). We describe the knowledge and attitudes of these HCPs, and identify modifiable barriers to appropriate anti-tuberculosis treatment. METHODOLOGY: We surveyed HCPs practicing in 30 clinical settings (hospitals, community health centers, and health posts) in the San Juan de Lurigancho district of Eastern Lima, Peru. Multiple-choice questions were used to assess knowledge of TB. A five-item Likert scale was created to assess attitudes toward the community, patients, and clinics. Linear regression was used to identify predictors of mean knowledge score, and analysis of variance was used to test differences in HCP score. RESULTS: Of the 73 HCPs surveyed, 15% were professionals (doctors or nurses). The remaining 85% were health technicians, community health workers (CHWs) or students. The mean knowledge score was 10.0 +/- 1.9 (maximum 14) with professional HCPs scoring higher than other HCPs (11.7 +/- 1.1 vs. 9.7 +/- 1.9), p < .01). Knowledge gaps included identification of patients at high risk for TB, assessment of treatment outcomes, and consequences of treatment failure. The most commonly cited modifiable barriers were structural, including laboratory facilities and staffing of TB clinics, with 52.1% and 62.5% of HCPs, respectively, citing these as problematic. CONCLUSIONS: Efforts to improve knowledge of TB HCPs in Peru should focus on the specific gaps we have identified. Further research is needed to evaluate whether these knowledge gaps correlate with TB control. PMID- 20009281 TI - Frequency of scabies in Iraq: survey in a dermatology clinic. AB - BACKGROUND: The burden of scabies is highest in tropical countries, but recent data from Iraq are scanty. OBJECTIVE: To estimate the frequency of scabies in patients of a dermatology clinic in central Iraq. METHODOLOGY: The study was conducted in Tikrit (Iraq), between May 2007 and February 2008. A total of 1,194 patients presenting to a dermatology clinic were included in the study. RESULTS: During the study period, 132 [1.1%] patients with scabies were identified. The frequency was 13.5% in males and 8.6% in females [P = 0.007]. Frequency in children was 15.6% and 9.8% in adults [P = 0.007]. About 91% of cases were younger than 45 years. The treatment of scabies cases with 5% permethrin cream resulted in a cure rate of 80.3% following a single application, and was increased to 95.5% after a second application. Cure was achieved in all cases after a third application. CONCLUSION: Prevalence of scabies was high in a dermatology clinic, and prisoners were identified as high-risk group. Mass treatment of scabies either by oral ivermectin or topical permethrin is suggested. PMID- 20009282 TI - Transfusion-transmissible infections among blood donors in Kathmandu, Nepal. AB - BACKGROUND: Screening of transfusion-transmissible infections (TTIs) among blood donors can be a cost-effective approach to monitor the prevalence, distribution, and trends of the infections among healthy-looking individuals. The study aimed to determine the seroprevalence of four TTIs, human immunodeficiency virus (HIV), hepatitis B virus (HBV), hepatitis C virus (HCV) and syphilis, among blood donors in Kathmandu, Nepal. METHODOLOGY: A total of 21,716 units of blood were tested for the presence of anti-HIV 1/2 IgG/IgM, HBsAg, anti-HCV IgG/IgM, and anti Treponema pallidum IgG/IgM/IgA using commercial ELISA kits following standard protocols. Statistical analysis was performed using WinPepi Ver 3.8. RESULTS: Seroprevalence of HIV, HBV (HBsAg), HCV and syphilis were observed to be 0.12% (95% CI = 0.08-0.18), 0.47% (95% CI = 0.39-0.57), 0.64% (95% CI = 0.54-0.75) and 0.48% (95% CI = 0.40-0.59) respectively. TTIs were dominant among male blood donors compared to female blood donors. Higher HCV seroprevalence among males compared to females was statistically significant. HIV prevalence was highest among blood donors in the age group 31 to 40 years (P > 0.5). HBV, HCV and syphilis prevalence was highest among blood donors 41 to 50 years age group, 21 to 30 years age group, and 51 to 60 years age group respectively (P < 0.05). HIV and HBV prevalence was relatively higher among first-time donors, whereas HCV and syphilis was relatively higher among the repeated donors (P > 0.05). CONCLUSIONS: It is of utmost importance to continue screening donated blood with highly sensitive and specific tests and to counsel donors who are positive to any of the above infections. It is absolutely necessary to avoid the transmission of infection from repeat donors. PMID- 20009283 TI - Trends in antimicrobial resistance in Shigella species in Karachi, Pakistan. AB - BACKGROUND: Shigellosis is a common cause of morbidity, especially in the very young and old, in developing countries. The disease is treated with antibiotics. Surveillance of antimicrobial resistance trends is essential owing to the global emergence of antimicrobial resistance. METHODOLOGY: The study involved 1,573 isolates of Shigella species (1996-2007) that were analyzed for trends in antimicrobial resistance. RESULTS: The majority of the specimens (1046; 66.5%) were from the pediatric population, and of these 887 (84.8%) were under 5 years of age (p = 0.001). S. flexineri was the most frequent species (54.5%) isolated. Isolation of S. sonnei increased from 15.4 % (1996) to 39% (2007) (p = 0.001). Although none of the isolates was found sensitive to all the antibiotics tested, 58% (n =9 07) were resistant to ampicillin and 85% (n = 1,338) were resistant to trimethoprim-sulfamethoxazole (TMP-SMX). Out of a total of 198 (12.6%) nalidixic acid resistant isolates, 6 (3.0%) were also resistant to ofloxacin. Overall 1.7 % of isolates were resistant to ofloxacin, 2.4% to ceftriaxone and 2.3% were resistant to combination of ampicillin, nalidixic acid and TMP-SMX. CONCLUSION: Ofloxacin is still an effective drug for treatment of acute shigellosis in Pakistan. Emergence of resistance to ceftriaxone in Shigella may have grave implications in treatment of severe shigellosis in very young patients. PMID- 20009284 TI - Association between HSV-2 and HIV serostatus in pregnant women of known HIV serostatus attending Mulago hospital antenatal clinic, Kampala, Uganda. AB - BACKGROUND: Studies show that STIs such as HSV-2 increase the probability of HIV 1 acquisition and enhance transmission by increasing susceptibility and infectiousness respectively. The objective of this study was to compare the proportion of HSV-2 in HIV-positive and HIV-negative pregnant mothers attending the antenatal clinic in Mulago Teaching Hospital in Kampala, Uganda. METHODS: This case control study included 50 pregnant women who were HIV positive and 200 controls of pregnant women who were HIV negative and were aware of their serostatus. Data was collected in two parts: the first part involved a pretested interviewer-administered semi-structured questionnaire for socio-demographic characteristics, sexual and behavioral history. The second part consisted of a serological test using an ELISA assay specific for IgG against viral glycoprotein G, specific to HSV-2. RESULTS: The proportion of HSV-2 was 86% (43/50) among the HIV-positive cases and 62% (125/200) among the HIV-negative controls. The odds of being HSV-2 seropositive was 3.7 times higher (95% CI was 1.58 - 8.61) in HIV positive cases than in the HIV negative controls. The odds of HSV-2 was significantly increased to 5.32 (95 CI was 1.92 - 14.73) among cases when adjustment was done for age, education, marital status, religion, age at first sexual experience, lifetime partners, type of earning, and whether the mother involved the partner in seeking treatment for sexually transmitted diseases. CONCLUSION: The proportion of HSV-2 was higher in HIV-positive cases than in the HIV negative controls. PMID- 20009285 TI - Hydatid cyst of the cranial vault. AB - Only 0.5 to 2% of hydatid cysts are localized in the skeleton and of these, 3 to 4% are found in the skull. In this paper, the authors report a case of primary hydatidosis involving the cranial vault revealed by a bulging mass of the forehead and symptoms of raised intracranial pressure that occurred in a 22-year old woman who came from a rural area. Through this case and literature review, the authors analyse the epidemiological, clinical and radiological aspects of skull hydatidosis. They conclude that hydatid cyst should be considered in the differential diagnosis of any soft tissue swelling or osteolytic lesion in the scalp of patients living in endemic areas. PMID- 20009286 TI - Tumors show enhanced dependency on glucose and glycolytic pathway. PMID- 20009287 TI - Enhancement of radiation and chemotherapeutic drug responses by 2-deoxy-D-glucose in animal tumors. AB - The development of an approach based on the energy-linked modification of DNA repair and cellular recovery processes using 2-deoxy-D-glucose (2-DG; inhibitor of glycolytic ATP production) has shown promising results in a number of model systems of cancer. Following encouraging results on the tolerance and toxicity (acute as well as late effects) of the combination (2-DG and hypofractionated radiotherapy) in Phase I and II clinical trials, its efficacy is currently under evaluation in Phase III clinical trials for glioma patients. Since heterogeneous physiologic and metabolic status in tumors as well as host-tumor interactions influence the local tumor control, which coupled with systemic disturbances could determine the cure (long-term tumor free survival), investigations on the in vivo responses of tumors to the combined treatment have received considerable attention. This communication provides a brief overview on the in vivo studies related to radio- and chemosensitization of tumors by 2-DG, besides the normal tissue toxicity induced by the combined treatment of 2-DG and radiation or chemotherapeutic drugs. PMID- 20009289 TI - Clinical studies for improving radiotherapy with 2-deoxy-D-glucose: present status and future prospects. AB - Higher rates of glucose usage generally correlate with poor prognosis in several types of malignant tumours. Experimental studies (both in vitro and in vivo) have shown that 2-deoxy-D-glucose (2-DG), a glucose analog and glycolytic inhibitor, enhances radiation-induced damage selectively in tumor cells while protecting normal cells, thereby suggesting that 2-DG can be used as a differential radiomodifier to improve the efficacy of radiotherapy. Clinical trials undertaken to study the feasibility, safety, and validity of this suggested approach will be described. Based on 2-DG-induced radiosensitization observed in primary organ cultures of cerebral glioma tissues, clinical trials were designed taking into consideration the radiobiology of gliomas and pharmacokinetics of 2-DG. Phase I/II clinical trials have unequivocally demonstrated that a combination of 2-DG (200-300 mg 2-DG per kg body weight orally administered after overnight fasting, 20 min before irradiation) with large weekly fractions (5 Gy/fraction) of low-LET radiotherapy is well tolerated without any acute toxicity or late radiation damage to the normal brain tissue. Nonserious transient side effects similar to hypoglycemia induced disturbances like restlessness, nausea, and vomiting were observed at the 2-DG doses used. Data from these trials involving more than 100 patients have clearly indicated a moderate increase in the survival, with a significant improvement in the quality of life with clinicopathological evidence of protection of normal brain tissue. A phase III multicentric trial to evaluate the efficacy of the combined treatment is in progress. Directions for future studies are discussed. PMID- 20009290 TI - Cytotoxicity, radiosensitization, and chemosensitization of tumor cells by 2 deoxy-D-glucose in vitro. AB - The glucose analog 2-deoxy-D-glucose (2-DG), an inhibitor of glucose transport and glycolytic ATP production, is the most widely investigated metabolic inhibitor for targeting glucose metabolism. Besides depleting energy in cells, 2 DG has also been found to alter N-linked glycosylation leading to unfolded protein responses and induce changes in gene expression and phosphorylation status of proteins involved in signaling, cell cycle control, DNA repair, calcium influx, and apoptosis. Inhibition of cell proliferation and induction of apoptosis have been observed as cytotoxic effects in a wide variety of tumor cells in vitro, while sensitization of tumor cells to ionizing radiation and certain chemotherapeutic drugs is associated with enhanced mitotic as well as apoptotic cell death induced by the primary therapeutic agent. Therefore, there has been a considerable amount of interest in developing 2-DG as a therapeutic agent or adjuvant in the radiotherapy and chemotherapy of tumors. PMID- 20009288 TI - Glucose deprivation-induced metabolic oxidative stress and cancer therapy. AB - Cancer cells (vs. normal cells) demonstrate evidence of oxidative stress, increased glycolysis, and increased pentose cycle activity. The oxidative stress in cancer cells has been hypothesized to arise from mitochondrial dysfunction leading to increased levels of hydroperoxides, and cancer cells have been proposed to compensate for this defect by increasing glucose metabolism. Glucose metabolism has also been shown to play a role in hydroperoxide detoxification via the formation of pyruvate (from glycolysis) and NADPH (from the pentose cycle). Furthermore, in cancer cells, glucose deprivation as well as treatment with 2 deoxyglucose (2 DG) has been shown to induce oxidative stress and cytotoxicity. Additionally, transformed cells have been shown to be more susceptible to glucose deprivation (and 2DG-)-induced cytotoxicity and oxidative stress than untransformed cells. These results support the hypothesis that cancer cells have a defect in mitochondrial respiration leading to increased steady state levels of O2*- and H2O2, and glucose metabolism is increased to compensate for this defect. The application of these findings to developing cancer therapies using 2DG combined with inhibitors of hydroperoxide metabolism to induce radio/chemosensitization is discussed, as well as the possibility that FDG-PET imaging may predict tumor responses to these therapies. PMID- 20009291 TI - Protection of normal cells and tissues during radio- and chemosensitization of tumors by 2-deoxy-D-glucose. AB - Normal tissue toxicity is one of the major limiting factors in cancer therapy. Damage to normal tissues and critical organs restricts the use of higher therapeutic doses thereby compromising the efficacy. The glucose analog 2-deoxy-D glucose (2-DG), an inhibitor of glycolytic ATP production has been shown to enhance radiation- and chemotherapeutic drug-induced damage in a number of cancer cells under in vitro and in vivo conditions while sparing or protecting normal cells. This review summarizes current understanding on the protection of normal cells and tissues against radiation- and chemotherapeutic drug-induced damage by 2-DG that makes this glucose analog an ideal adjuvant in cancer therapy. PMID- 20009292 TI - Different ways to improve the clinical effectiveness of radioimmunotherapy in solid tumors. AB - Radioimmunotherapy (RIT) has been proven effective in the treatment of radiosensitive non-Hodgkin lymphoma but, for radioresistant solid tumors, new approaches are necessary to improve the clinical effectiveness. A real improvement has been the introduction of the pretargeting technology which appeared to be able to significantly increase tumor-to-normal organ uptake ratios.Another very promising approach consists in associating RIT with other treatment modalities. Finally the use of alpha particle-emitting radionuclides such as astatin-211 or bismuth-213 (alpha-RIT) should allow to efficiently eradicate disseminated microscopic clusters of tumor cells or isolated tumor cells which fit well with the short path length of alpha particles. PMID- 20009293 TI - Chemosensitizing and cytotoxic effects of 2-deoxy-D-glucose on breast cancer cells. AB - BACKGROUND: Accelerated glucose uptake for anaerobic glycolysis is one of the major metabolic changes found in malignant cells. This property has been exploited for imaging malignancies and as a possible anticancer therapy. The nonmetabolizable glucose analog 2-deoxyglucose (2 DG) interferes with glucose metabolism leading to breast cancer cell death. AIMS: To determine whether 2DG can synergize with chemotherapeutic agents commonly used in breast cancer treatment and identify cellular characteristics associated with sensitivity to 2DG. MATERIALS AND METHODS: SkBr3 breast cancer cells were incubated with varying concentrations of 5-fluorouracil (5FU), doxorubicin, cisplatin, cyclophosphamide, or herceptin with or without 2DG. Cell viability was measured using the MTT (3 (4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay. RESULTS: Combining 2DG with doxorubicin, 5 FU, cyclophosphamide, and herceptin resulted in enhanced cell death compared with each agent alone, while in combination with cisplatin, the amount of cell death was additive. Mouse embryo fibroblasts (MEF) mutated for p53 (-/-) were 30% more sensitive to the cytotoxic effects of 2DG than the parental cell lines. Cells mutated for Bax/Bac, genes involved in protection from apoptosis, are slightly more sensitive than the parental cell lines. CONCLUSIONS: These results indicate that 2DG acts synergistically with specific chemotherapeutic agents in causing cell death and the class of chemicals most sensitive appear to be those which cause DNA damage. PMID- 20009294 TI - Differential responses of tumors and normal brain to the combined treatment of 2 DG and radiation in glioablastoma. AB - 2-Deoxy-D-glucose (2-DG), an inhibitor of glucose transport and glycolysis, enhances radiation damage selectively in tumor cells by modulating damage response pathways resulting in cell death in vitro and local tumor control. Phase I and II clinical trials in patients with malignant glioma have shown excellent tolerance to a combined treatment of orally administered 2-DG and hypofractionated radiotherapy without any acute toxicity and late radiation damage. Phase III efficacy trials are currently at an advanced stage. Re exploratory surgery performed in 13 patients due to persistent symptoms of elevated ICP and mass effect at different follow-up periods revealed extensive tumor necrosis with well-preserved normal brain tissue adjoining the tumor included in the treatment volume as revealed by a histological examination. These observations are perhaps the first clinical evidences for differential effects of 2-DG on tumors and normal tissues in conformity with earlier in vitro and in vivo studies in normal and tumor-bearing mice. PMID- 20009295 TI - Modification of 2-deoxy-D-glucose on radiation-and chemotherapeutic drug-induced chromosomal aberrations. AB - BACKGROUND: Chemotherapy is the treatment of cancer with drugs, often used as either adjuvant or neoadjuvant or in conjunction with radiation and surgery. Unfortunately, majority of the drugs are toxic to normal tissues, the toxicity being resulting from multidrug protocol used to induce remissions and achieve tumor care. While it has been demonstrated for compounds like the 2-deoxy-glucose (2-DG) used as a modulator for radiation-induced damages, such studies were rarely reported for chemotherapeutic drugs. OBJECTIVE: To study the effect of 2 DG on radiation-and chemotherapeutic drug-induced chromosomal aberrations in normal and tumor cells exposed in vitro. MATERIALS AND METHODS: The peripheral blood lymphocytes (PBLs) and BMG-1 cells were exposed to radiation and chemotherapeutic drugs (bleomycin and mitomycin-C) in the presence and absence of 2-DG. The treated cells were cultured for various durations, arrested at either metaphase or cytokinesis stage of the cell cycle. The stable and unstable aberrations were recorded using Giemsa staining and FISH technique. The cell cycle kinetics was studied using fluorescence plus Giemsa (FPG) staining. RESULTS: The presence of 2-DG reduced stable and unstable chromosome aberrations (CA) significantly (P < 0.001), in PBLs induced by radiation, bleomycin and mitomycin-C, when compared to cells treated with radiation or the drugs and increased significantly in BMG cells (P < 0.001). Furthermore, the presence of 2 DG altered the cell cycle kinetics in the PBLs and BMG-1 cells. Thus the overall results showed protection effect on the normal cell damages induced by radiation and chemotherapeutic drugs, while sensitizes the tumor cell. CONCLUSION: The obtained results suggest that 2-DG in combination with radiotherapy/chemotherapy could lead to an improvement in tumor therapy by sensitizing the tumor cells while protecting the normal cells. PMID- 20009296 TI - Modulatory action of 2-deoxy-D-glucose on mitomycin C-and 4-nitroquinoline-1 oxide-induced genotoxicity in Swiss albino mice in vivo. AB - BACKGROUND: 2-Deoxy-D-glucose (2-DG), a structural analog of glucose is an effective inhibitor of glucose metabolism and ATP production. It selectively accumulates in cancer cells and interferes with glycolysis leading to cell death. 2-DG is shown to differentially enhance the radiation-induced damage in cancer cells both under euoxic and hypoxic conditions. A combination of 2-DG and ionizing radiation selectively destroys tumors while protecting the normal tissue. 2-DG is being advocated as an adjuvant in the radiotherapy and chemotherapy of cancer. OBJECTIVE: The present investigation focuses on the modulatory effect of 2-DG on mitomycin C- (MMC) and 4-nitroquinoline-1-oxide (4 NQO)-induced cytogenetic damage in bone marrow cells of Swiss albino mice in vivo. MATERIALS AND METHODS: Experimental animals were pretreated with 2-DG (500 mg/kg, i.p.) for five consecutive days followed by MMC (2 mg/kg, i.p) or 4-NQO (15 mg/kg, i.p.), 24 h prior to sacrifice. Control animals were given either the mixture of olive oil and acetone (3:1) or distilled water. Bone marrow cells were processed for the micronucleus assay and metaphase analysis for estimating cytogenetic damage. RESULTS: 2-DG significantly (P < 0.001) reduced the frequency of aberrant cells induced by MMC (approximately 90%) and 4-NQO (approximately 74%). Incidence of micronucleated polychromatic erythrocytes (MnPCEs) induced by the mutagens were reduced up to 68%. CONCLUSION: 2-DG effectively reduces the MMC and 4-NQO-induced genotoxicity. PMID- 20009297 TI - Modulation of cellular radiation responses by 2-deoxy-D-glucose and other glycolytic inhibitors: implications for cancer therapy. AB - BACKGROUND: 2-Deoxy-D-glucose (2-DG), a glycolytic inhibitor, was observed earlier to increase DNA, chromosomal, and cellular damage in tumor cells, by inhibiting energy-dependent repair processes. Lonidamine (LND) selectively inhibits glycolysis in cancer cells. It damages the condensed mitochondria in these cells, impairing thereby the activity of hexokinase (predominantly attached to the outer mitochondrial membranes). It inhibits repair of radiation-induced potentially lethal cellular damage in HeLa and Chinese hamster (HA-1) cells. However, other than a preliminary study on human glioma (BMG-1) cells in this laboratory, the effects of LND on radiation-induced cytogenetic damage have not been reported earlier. AIMS: These studies were carried out to investigate the effects of LND and 2-DG on cell proliferation, viability, and radiation response in the same human glioma cell line, under identical conditions. The respective drug concentrations were selected on the basis of earlier studies. MATERIALS AND METHODS: Human glioma (U373MG) cells were grown in the presence of LND or 2-DG for 2 days. Proliferation response and viability of U373MG human glioma cells were studied by cell counts and uptake of trypan blue dye. Radiosensitization (increase in micronuclei formation) was studied after short-term (4 h postirradiation) drug treatments. OBSERVATIONS: Both the drugs (1) inhibited proliferation response in a concentration-dependent manner; (2) did not induce micronuclei formation in the unirradiated cells; and (3) significantly increased radiation-induced micronuclei formation at nontoxic concentrations. CONCLUSIONS: These data suggest that the short-term presence of either lonidamine or 2-DG-at clinically relevant and nontoxic concentrations-could increase the treatment response of malignant gliomas at optimum radiation doses, reducing thereby the side effects of radiotherapy. PMID- 20009298 TI - Association between the unfolded protein response, induced by 2-deoxyglucose, and hypersensitivity to cisplatin: a mechanistic study employing molecular genomics. AB - BACKGROUND: The specific signaling that occurs between the endoplasmic reticulum (ER) and the nucleus in response to ER stress is known as the unfolded protein response (UPR). Specific induction of GRP78 (glucose-regulated protein of Mr 78 kDa) is an integral component of ER stress and the UPR. We first discovered that the up-regulation of GRP78 is associated with augmented sensitivity/apoptosis of cancer cells to clinically used alkylating/platinating agents. OBJECTIVES: To decipher molecular mechanisms that associate induction of the UPR/GRP78 with augmented sensitivity/apoptosis to cisplatin. MATERIALS AND METHODS: A549 cells were exposed to 2-deoxyglucose (2dG) to induce the UPR/GRP78, followed by cisplatin treatment. We used human cDNA microarray containing 42,000 ESTs as well as pathway-specific macroarrays for apoptosis, cell cycle, and MAP kinase signaling pathways containing 100-280 genes and subsequently examined the pertinent transcript levels. The results obtained from these studies were confirmed by examining relevant protein levels and the enzymatic activity. RESULTS: We demonstrate that the induction of UPR/GRP78 alone causes a decrease in the transcript levels of DNA repair genes and DNA damage check point genes, and an increase in the transcript levels of apoptotic genes. Furthermore, we show that cisplatin treatment after the induction of UPR/GRP78 is facilitating the mitochondria-mediated apoptotic cascades through the initial activation of caspase-2 and down-regulation of genes involved in DNA repair. CONCLUSIONS: Our study will shed new insight as to the increased understanding of the mechanisms of the UPR/GRP78 modulation of molecular and cellular responses to cisplatin that will allow strategies for transferring bench side results to the bed. PMID- 20009299 TI - Short-term exposure of multicellular tumor spheroids of a human glioma cell line to the glycolytic inhibitor 2-deoxy-D-glucose is more toxic than continuous exposure. AB - The glycolytic inhibitor 2-deoxy-D-glucose (2-DG) has been used as a therapeutic agent and as an adjuvant in cancer therapy with either weekly fractions of the treatment or daily administration. While the weekly fraction has often been found to be nontoxic and effective, other treatment regimes are tolerated to a relatively lesser extent. It was therefore, considered worthwhile to investigate the efficacy of short- and long-term exposure of tumor cells to 2-DG under the controlled conditions. Seven-day-old MTS were exposed to 2-DG (5 mM, equimolar to glucose concentration in media) for different time intervals (30 min to 24 h) trypsinized and plated for clonogenicity. Alternatively, spheroids were grown either continuously in the presence of 2-DG or were treated with 2-DG for 2 h (short-term exposure) and grown in 2-DG-free media for 21 days and assessed for spheroid growth, cell viability, apoptosis, cytogenetic damage, mitochondrial status, and oxidative stress. Exposure of spheroids to 2-DG for 2-4 h induced 30% cell death (SF 0.70) while, a 24-h exposure resulted in only a marginal decrease in clonogenicity (SF 0.95). Furthermore, the spheroids disintegrated completely by 28 days in the case of 2-h exposure to 2-DG, while spheroids grown continuously in the presence of 2-DG repopulated. The cytotoxicity following short-term exposure of MTS to 2-DG was primarily due to the induction of apoptosis revealed by morphological features as well as flow cytometric analysis of the DNA content. Interestingly however, cytogenetic damage (micronuclei induction) was observed in spheroids that were continuously exposed to 2-DG. Short-term exposure to 2-DG resulted in a significant increase in ROS levels and a reduction in the levels of unoxidized cardiolipin as measured by NAO suggesting the involvement of mitochondria leakiness leading to oxidative stress which, could be responsible for apoptotic cell death observed under these conditions. However, continuous exposure to 2-DG resulted in a moderate level of oxidative stress leading to the genomic instability. Preliminary studies also show that spheroids exposed continuously to 2-DG result in the development of resistance to certain chemotherapeutic drugs which could be correlated with elevated levels of mdr1. The present results suggest that a persistent down-regulation of glycolysis (as seen here with continuous exposure to 2-DG) could activate prosurvival responses besides inducing moderate levels of oxidative stress resulting in the development of resistance against therapeutic agents. PMID- 20009300 TI - Targeting energy metabolism in brain cancer through calorie restriction and the ketogenic diet. AB - Malignant brain tumors are a significant health problem in children and adults and are largely unmanageable. As a metabolic disorder involving the dysregulation of glycolysis and respiration (the Warburg effect), malignant brain cancer can be managed through changes in metabolic environment. In contrast to malignant brain tumors that are mostly dependent on glycolysis for energy, normal neurons and glia readily transition to ketone bodies (beta-hydroxybutyrate) for energy in vivo when glucose levels are reduced. The transition from glucose to ketone bodies as a major energy source is an evolutionary conserved adaptation to food deprivation that permits the survival of normal cells during extreme shifts in nutritional environment. Only those cells with a flexible genome, honed through millions of years of environmental forcing and variability selection, can transition from one energy state to another. We propose a different approach to brain cancer management that exploits the metabolic flexibility of normal cells at the expense of the genetically defective and less metabolically flexible tumor cells. This approach to brain cancer management is supported from recent studies in orthotopic mouse brain tumor models and in human pediatric astrocytoma treated with calorie restriction and the ketogenic diet. Issues of implementation and use protocols are discussed. PMID- 20009301 TI - What is new in emergencies, trauma and shock? Studying stress in emergency medicine. PMID- 20009302 TI - Angiotensin-converting enzyme gene polymorphism in hypertensive rural population of Haryana, India. AB - BACKGROUND: Essential hypertension is a complex genetic disorder influenced by diverse environmental factors. Of the various physiological pathways affecting the homeostasis of blood pressure, the renin-angiotensin system (RAS) is known to play a critical role. Angiotensin-I converting enzyme (ACE) is a significant component of RAS and an insertion/deletion (I/D) polymorphism in its gene has been implicated in predisposition to hypertension. OBJECTIVE: The present study is aimed to determine the association, if any, of ACE I/D polymorphism with essential hypertension in a rural population of Haryana, India. MATERIALS AND METHODS: The blood samples were collected from the patients visiting M. M. Institute of Medical Sciences, Mullana, Haryana. DNA from the patients (106) and control (110) specimens were isolated, amplified by PCR and analyzed employing agarose gel electrophoresis. RESULTS: There was no significant difference in the distribution of DD, II and I/D genotypes of ACE polymorphism in essential hypertensive patients (28.8, 25.5, and 46.2%) and their ethnically matched normal control (24.5, 30, and 45.5), respectively. The two groups also presented with very similar allelic frequencies and were also found to be in Hardy-Weinberg equilibrium. CONCLUSIONS: The present study demonstrates that ACE I/D polymorphism is not a risk factor for essential hypertension in the hitherto unstudied rural population of Haryana. PMID- 20009303 TI - A cross-sectional study of the self-report of stress among emergency department patients. AB - BACKGROUND: There is a lack of information about the prevalence of stress and types of stressors experienced by Emergency Department (ED) patients. OBJECTIVE: The present aim is to study the prevalence of stress, types of stressful situations and the relationship with other health issues within the ED population. MATERIALS AND METHODS: This is a cross-sectional study performed in an ED at a level-1 urban trauma center for four months. An anonymous survey was offered to adult non critically ill patients who were admitted in the ED. They were divided on the basis of gender, age and level of stress and were asked about their demographics, reasons for their ED visit and health issues including stress, tobacco, alcohol and marijuana use, weight concern and health. Chi-square for the categorical variables and unpaired t-tests for continuous variables were conducted. RESULTS: We interviewed 1797 patients, over 66% reported that they felt stressed on at least a weekly basis, and over 45% indicated that they felt stressed more than twice per week. While both young and old were equally stressed, females reported significantly more stress. Family, finances and work are the most frequently cited stressors. Different age groups reported different types of stressors. Overall, those patients reporting being stressed more frequently reported more high risk behaviors including cigarette and marijuana use and health problems including being overweight, being depressed, and having sleeping problems and chronic fatigue. CONCLUSIONS: Considering the high frequency of ED patients that report frequent stress and high risk behaviors, their ED visit may be an excellent opportunity to provide a referral or an intervention for stress reduction. PMID- 20009304 TI - Etiological characterization of acute poisonings in the emergency department. AB - INTRODUCTION: An investigation of emergency department (ED) poisonings was conducted to characterize poisoning demographics and evaluate correlations with select co-morbidities. METHODS: The study population evaluated consisted of 649 poisoning cases admitted between 2004 and 2007 to an inner-city, level 1 emergency department. RESULTS: Ethnicity, age, and gender had a substantial impact on the population distribution as poisoning cases were predominantly African Americans (79.9%) between 36 and 45 years old with a 1:3 male to female ratio. Intentional illicit drug overdose was the most prevalent cause of poisoning, heroin being the most frequent substance found in 35.4% ( n = 230) of cases, followed by cocaine overdose at 31.7% ( n = 206), concomitant heroin and cocaine overdose at 4.3% ( n = 28), multiple drug poisoning at 5.5% ( n = 36), and antidepressant/antipsychotic poisoning at 6% ( n = 39). Significant correlations were found between heroin poisoning and asthma (F = 20.29, DF = 1, P = 0.0001), cocaine poisoning and hypertension (F = 33.34, DF = 1, P = 0.0001), and cocaine poisoning and cardiovascular disease (F = 35.34, DF = 1, P = 0.0001). A change in the pattern of illicit drug use from injection to inhalation was detected and the resulting increase of inhalation and insufflation of illicit substances may partially explain the correlation found between heroin use and asthma. CONCLUSIONS: These results provide supporting evidence that deliberate poisoning with illicit drugs remains a serious healthcare issue that significantly aggravates co-morbidities and raises treatment costs by increasing both the rate of hospitalization and hospital length of stay. PMID- 20009305 TI - Comparison of two different types of heat and moisture exchangers in ventilated patients. AB - STUDY OBJECTIVES: To compare the efficacy of two different types of Heat and Moisture Exchangers (HME filters) in reducing transmission of infection from the patient to ventilator and vice versa and also its cost effectiveness. DESIGN: Randomized, controlled, double blind, prospective study. PATIENTS AND METHODS: 60 patients admitted to the ICU from May 1, 2007 to July 31, 2007 of either sex, age ranging between 20 and 60 years, requiring mechanical ventilation were screened for the study. Following intubation of the patients, the HME device was attached to the breathing circuit randomly by the chit-in-a box method. The patients were divided into two groups according to the HME filters attached. RESULTS: Both the groups were comparable with respect to age and sex ratio. In Type A HME filters, 80% showed growth on the patient end within 24 h and in 27% filters, culture was positive both on the patient and the machine ends. The organisms detected were Staphylococcus aureus, Escherichia coli and Pseudomonas aeruginosa and co-related with the endotracheal aspirate culture. After 48 h, 87% filters developed organisms on the patient end, whereas 64% filters were culture positive both on the patient and the machine end. In Type B HME filters, 70% showed growth on patient's end after 24 h. Organisms detected were S. aureus, E. coli, P. aeruginosa and Acinetobacter. Thirty percent of filters were culture negative on both the patient and machine ends. No growth was found on the machine end in any of the filters after 24 h. After 48 h, 73% of the filters had microbial growth on the patient end, whereas only 3% filters had growth (S. aureus) on the machine end only. Seven percent had growth on both the patient as well as the machine ends. The microorganisms detected on the HME filters co-related with the endotracheal aspirate cultures. CONCLUSION: HME filter Type B (study group) was significantly better in reducing contamination of ventilator from the patient as compared to Type A (control group), which was routinely used in our ICU. Type B filter was found to be effective for at least 48 h. This study can also be applied to patients coming to emergency department (ED) and requiring emergency surgery and postoperative ventilation; and trauma patients like flail chest, head injury etc. requiring ventilatory support to prevent them from acquiring ventilator-associated pneumonia (VAP). PMID- 20009306 TI - Emergency medical epidemiology in Assam, India. AB - BACKGROUND: Assam, with its capital in Dispur has one of the highest rates of infant and maternal mortality in India. Being under both tribal and hilly regions, it has lacked adequate healthcare and emergency services. We therefore aimed to conduct a cross-sectional survey of medical emergencies and identify various types of emergencies presenting to emergency departments, prior to launching emergency services across the state. MATERIALS AND METHODS: On a prospective basis and using a stratified random sampling design, all emergencies presenting to the three government hospitals in Guwahati, Assam, which handle 90% of all emergencies currently, were studied on specially designed datasheets in order to collect data. Emergency medical technicians (EMTs) were placed in the Casualty of the medical colleges and recorded all emergencies on the datasheet. The collected data was then analysed for stratification and mapping of emergencies. In addition, retrospective data for a period of 15 days was collected from the emergency case registers of all three hospitals and the adjoining district civil hospitals, in order to give a wider perspective of the nature of emergencies. RESULTS: A total of 2169 emergencies were recorded over a seven-day prospective and fifteen-day retrospective period. Guwahati Medical College Hospital attended to majority of emergencies (42%), which were mainly of the nature of pregnancies (22.7%), accidents (12.2%) or assaults (15.4%) and fever related. Maximum emergencies also presented from the border districts, and occurred among young males in the age group of 19-45 years. Males were also more prone to accidents and assaults, while females presented with pregnancies as emergencies. CONCLUSION: Potential emergency services need to target young pregnant females. Law and order needs to be also tightened in order to curb accidents and assaults among young males. PMID- 20009307 TI - Hospital design for better infection control. AB - The physical design and infrastructure of a hospital or institution is an essential component of its infection control measure. Thus is must be a prerequisite to take these into consideration from the initial conception and planning stages of the building. The balance between designing a hospital to be an open, accessible and public place and the control to reduce the spread of infections diseases is a necessity. At Singapore General Hospital, many lessons were learnt during the SARS outbreak pertaining to this. During and subsequent to the SARS outbreak, many changes evolved in the hospital to enable us to handle and face any emerging infectious situation with calm, confidence and the knowledge that staff and patients will be in good stead. This paper will share some of our experiences as well as challenges. PMID- 20009308 TI - Acute management of vascular air embolism. AB - Vascular air embolism (VAE) is known since early nineteenth century. It is the entrainment of air or gas from operative field or other communications into the venous or arterial vasculature. Exact incidence of VAE is difficult to estimate. High risk surgeries for VAE are sitting position and posterior fossa neurosurgeries, cesarean section, laparoscopic, orthopedic, surgeries invasive procedures, pulmonary overpressure syndrome, and decompression syndrome. Risk factors for VAE are operative site 5 cm above the heart, creation of pressure gradient which will facilitate entry of air into the circulation, orogenital sex during pregnancy, rapid ascent in scuba (self contained underwater breathing apparatus) divers and barotrauma or chest trauma. Large bolus of air can lead to right ventricular air lock and immediate fatality. In up to 35% patient, the foramen ovale is patent which can cause paradoxical arterial air embolism. VAE affects cardiovascular, pulmonary and central nervous system. High index of clinical suspicion is must to diagnose VAE. The transesophgeal echocardiography is the most sensitive device which will detect smallest amount of air in the circulation. Treatment of VAE is to prevent further entrainment of air, reduce the volume of air entrained and haemodynamic support. Mortality of VAE ranges from 48 to 80%. VAE can be prevented significantly by proper positioning during surgery, optimal hydration, avoiding use of nitrous oxide, meticulous care during insertion, removal of central venous catheter, proper guidance, and training of scuba divers. PMID- 20009310 TI - Managing a wooden foreign body in the neck. AB - An interesting case is presented of a wooden foreign body in the neck entering through the right lower vestibule of the mouth. The foreign body traveled subcutaneously in the neck and got stuck in the clavicle, without damaging any vital structures. In this case, the authors highlight the mode of entry of the foreign body, the peculiarity of the wooden foreign body, the management protocol and the outcomes of the penetrating neck injury by the wooden stick. PMID- 20009309 TI - Managing human bites. AB - Human bites are frequently overlooked in making a diagnosis in the emergency room. They are particularly notorious due to the polymicrobial nature of human saliva inoculated in the wound and the risk they pose for transmission of infectious diseases. Early treatment, appropriate prophylaxis and surgical evaluation are the key to achieving desired treatment outcomes. Through this article, we have tried to summarize the diagnostic features, complications as well as the recommended treatment alternatives for human bites based on the current available evidence. PMID- 20009311 TI - Abdominal pregnancy as a cause of hemoperitoneum. AB - The coexistence of intrauterine and extrauterine pregnancy, the heterotopic pregnancy, is a rare obstetric phenomenon. The preoperative diagnosis of this condition is very difficult; leading to a higher maternal morbidity and fetal loss. We experienced a case of intrauterine pregnancy and ruptured abdominal pregnancy implanted on the illeocaecal region in a 26-year-old primiparous woman. She was clinically misdiagnosed as a case of ruptured ectopic pregnancy, but ultrasonography showed it to be a case of heterotopic pregnancy. Subsequently, the patient was subjected to laparotomy and the ruptured abdominal pregnancy was evacuated. She continued with the intrauterine pregnancy till term and delivered a healthy female baby. Although this condition is unusual, any general surgeon in the emergency department must be aware of this complication and its management, which is often initially misdiagnosed. PMID- 20009313 TI - Wunderlich's syndrome and hemorrhagic shock. AB - We report a case of Wunderlich's syndrome in an obese woman associated with massive retroperitoneal hemorrhage. Stable hemodynamic patient condition was obtained by selective arterial embolization. Since the first embolization of a renal angiomyolipoma in 1976 by Moorhead et al., highly selective renal arterial embolization of angiomyolipoma with rupture has become a procedure that offers greater efficacy, particularly in life-threatening cases. PMID- 20009312 TI - Gastrointestinal stromal tumors and shock. AB - Gastrointestinal stromal tumors (GIST) are the most common mesenchymal tumor of the gastrointestinal tract. Clinically, they are associated with nonspecific symptoms, but some patients can present gastrointestinal bleeding with shock. We report two cases of GIST of the small bowel, revelated by hemorrhagic shock secondary to acute bleeding, succesfully treated by emergency surgery. PMID- 20009314 TI - Wellen's syndrome: An ominous EKG pattern. AB - Wellen's syndrome is a characteristic T-wave on an electrocardiogram during a pain-free period in a patient with intermittent chest pain. This finding suggests a high-degree stenosis of the proximal left anterior descending (LAD) coronary artery that will soon result in an acute anterior wall myocardial infarction (MI) if the patient is not urgently catheterized and the occlusion opened. This case report discusses a young male patient with no known cardiac disease with an EKG that demonstrates the classic Wellen's T-waves. He was urgently taken to cardiac catheterization and his 95% proximal LAD stenosis was reduced via drug-eluding stent. Through knowledge of Wellen's T-waves, more anterior wall MIs can be prevented. PMID- 20009315 TI - Periorbital swelling in emergency room: Get your eyes in. AB - Facial swelling and eye pain are very common patient complaints in Emergency Departments. Clinical evidence and investigations play a crucial role in making the correct diagnosis which impacts the final disposition and management of the patient. We present a case of a patient who presented with facial swelling and headache. PMID- 20009316 TI - Sudden unilateral visual field loss. AB - We report a classical case of branch retinal artery occlusion (BRAO) in the acute setting and review the literature relating to the diagnostic, therapeutic and prognostic facets of this condition. BRAO can cause sudden visual loss and is not an infrequent presentation to emergency medical services. BRAO may indicate predisposing and related conditions capable of significant morbidity and mortality. Although current therapeutic practices in the acute setting are of uncertain benefit, conservative measures may be attempted in the emergency room by a nonophthalmologist with the aim of dislodging the causative embolus. Regardless of the current means of acute management, anitplatelet therapy and cardiovascular risk management remain the mainstay of treatment for BRAO. The potential for life-threatening systemic associations necessities investigation and multidisciplinary input. PMID- 20009317 TI - Cut throat zone II neck injury and advantage of a feeding jejunostomy. AB - Penetrating neck injuries account for 5-10% of trauma cases and are potentially life threatening. We report a case of cut- throat zone II neck injury in a 45 year-old male extending up to posterior pharyngeal wall and exposing the underlying cervical vertebra. Tracheostomy was done and wound repair was started from the posterior aspect in layers using 3-0 Vicryl. Intraoperatively, a conscious decision was taken for a feeding jejunostomy for postoperative feeding, which was likely to be prolonged, in view of sensory-nerve damage along the transected pharynx. Prolonged use of Nasogastric tube for postoperative feeding was thus avoided and the discomfort, risk of aspiration and foreign body at injury site eliminated. One week postoperative, the patient experienced severe bouts of coughing and restlessness on oral intake; during this period enteral nutrition was maintained through feeding jejunostomy. At the time of discharge at 1 month, the patient was accepting normal diet orally and was detubated and vocalizing normally. We conclude that postoperative nutrition is an important area to be considered for deep neck wound with nerve injuries due to delayed tolerance to oral feeding till the regeneration of sensory nerves. A feeding jejunostomy or feeding gastrostomy performed simultaneously in such patients with nerve injuries is far superior over nasogastric-tube feeding when prolonged postoperative feeding is expected. PMID- 20009318 TI - Ileosigmoid knot strangles the sigmoid. PMID- 20009319 TI - Isolated transverse process fracture of the lumbar vertebrae. PMID- 20009320 TI - Emergency central venous catheterization revisited. PMID- 20009321 TI - Rolling circle amplification for signal enhancement in ovalbumin detection. AB - We have developed a rolling circle amplification-procedure as a method to enhance signals of immunoassay (immuno-RCA) to detect one of the major food allergens, ovalbumin (OVA). Immuno-RCA in combination with a fluorescent dye and a circular DNA probe prepared by intramolecular ligation easily allowed the real-time detection of OVA, and could apparently detect signals from OVA with concentrations ranging from 10(-12) to 10(-7) g/mL. Therefore, sensitive and easily handled aspects of this method would contribute to an effective signal enhancement in immunoassay for food allergen detections. PMID- 20009322 TI - 2-Aminotryptanthrin derivative with pyrene as a FRET-based fluorescent chemosensor for Al3+. AB - 1-(2-Tryptanthrinylaminoacetoxy)-14-(1-pyrenecarboxy)-3,6,9,12 tetraoxatetradecane (T2NH-P5P) was synthesized as a fluorescent chemosensor for Al3+. Excited at 325 nm, corresponding to absorption of the pyrene unit of T2NH P5P, emission at 600 nm from the 2-aminotryptanthrin unit has been observed, indicating that intramolecular fluorescence resonance energy transfer (FRET) occurs in T2NH-P5P. However, when Al3+ is added to a solution of T2NH-P5P, the fluorescence of 2-aminotryptanthrin is quenched (FRET-off), whereas that of the pyrene group is revived. Such a FRET "on-off" behavior of T2NH-P5P is not observed for other metal cations (Ca2+, Ba2+, and Zn2+). PMID- 20009323 TI - Facile method for constructing metallic nanoarrays on a solid surface. AB - This review highlights work using the author's method to construct metallic nanoarrays. This method is a DNA-templated assembly, to assemble metal nanoparticles (MNPs) in which DNA molecules are initially stretched and aligned on surfaces. Simple interface (air-liquid) moving and surface coating with polymers containing pi-conjugation units, lead to the controlled formation of one dimensional (1D) nanoarrays of DNA without special equipment. The assembly of MNPs onto DNA can be driven by electrostatic binding of newly prepared gold nanoparticles with positive charges, leading the formation of 1D metallic nanoarrays. Dark-field microscopy observations indicate that MNPs on DNA are uniaxially aligned. Finally, the fabrication and patterning of metallic nanoarrays achieved with DNA nanofibers and transfer printing techniques are discussed. PMID- 20009324 TI - Development of a micro-potentiometric sensor for the microchip analysis of alkali ions. AB - This paper reports on the development of a micro-potentiometric sensor based on external microelectrodes introduced into a microchip. We miniaturized reference and ion-selective electrodes (ISEs) and embedded them into a plastic (PDMS) microchip; the miniaturization of ISE was attained by using a monolithic capillary-based membrane. This sensor was applied to the detection of alkali ions (Na+, K+ and NH4+) in a microflow on the microg/L level. PMID- 20009325 TI - A microfluidic system to evaluate intestinal absorption. AB - Intestinal absorption rates vary with the nature of the substances involved. In vitro experiments with cell culture inserts are often conducted to evaluate the intestinal absorption rate. These inserts, however, require large amounts of cells, samples, and culture media, and take a long time to evaluate. To overcome these problems, we developed a microchip-based system that mimics the intestine. The microchip was composed of a glass slide, a permeable membrane, and polydimethylsiloxane (PDMS) sheets, which contained microchannels made by photolithography; Caco-2 cells were cultured on the membrane in the microchip. The system was regulated with a microsyringe pump. We conducted permeation tests; cyclophosphamide, which can permeate the intestinal barrier, displayed a high permeability coefficient and Lucifer yellow, which cannot be absorbed at the intestinal wall, displayed a low permeability coefficient. These results were consistent with those obtained using a conventional method, which supports the validity of our new system. The system realized an 80% reduction of cell consumption. PMID- 20009326 TI - A novel CdSe/CdS quantum dot-based competitive fluoroimmunoassay for the detection of clenbuterol residue in pig urine using magnetic core/shell Fe3O4/Au nanoparticles as a solid carrier. AB - Quantum dots (QDs) are semiconductor fluorescent nanoparticles, which can be used for food safety or environmental monitoring with high sensitivity. This work demonstrates the feasibility of detecting clenuterol residue in pig urine using CdSe/CdS quantum dots as fluorescent labels based magnetic core/shell Fe3O4/Au nanoparticles (MCFN) as solid carriers. The detection of clenbuterol is carried out by a fluoroimmunoassay-based biosensor using competitive binding between conjugated clenbuterol antigen-CdSe/CdS QDs and free clenbuterol with immobilized clenbuterol antibodies on MCFN. This assay method allows for clenbuterol determination in a linear working range of 0.5-20000 pg mL(-1). It would provide a simple, rapid, and ultra-sensitive detection method for clenbuterol or other biomolecular analysis. PMID- 20009327 TI - Determination of the androgenicity of ligands using a single-chain probe carrying androgen receptor N-terminal peptides. AB - The present study demonstrates a single-molecular bioluminescent probe carrying functional peptides in the N-terminal domain of the androgen receptor (AR NTD) with an improved sensorial property to androgens. The N-terminal peptides in AR were genetically fused to the ligand binding domain of AR (AR LBD) with a flexible linker, and then sandwiched between the N- and C-terminal fragments of split-firefly luciferase (FLuc) dissected at D415. We found that the proline-rich region in AR NTD efficiently interacts with AR LBD and exerts (i) an enhanced signal-to-background ratio and (ii) discrimination between agonists and antagonists with (iii) a 100-times improved sensitivity to androgens, upon comparison with previous references. A deletion mutation to the proline-rich region in AR revealed that this region is critical for the transcriptional activities. The quantum yields of these single-chain probes were estimated to be 37.8 +/- 0.6%. This monomeric AR LBD-peptide binding is necessary, and sufficient for discriminating an agonist and an antagonist, where the dimerization of AR LBD is not involved. The present study guides a fundamental methodology on how to discriminate weak protein-peptide binding, and provides a new insight into the contribution of functional peptides in AR NTD to the initial activation of monomeric AR. PMID- 20009328 TI - Glucose oxidase immobilized in alginate/layered double hydroxides hybrid membrane and its biosensing application. AB - A new type of amperometric glucose biosensor based on alginate (Alg)/layered double hydroxides (LDHs) organic-inorganic composite film is described. This hybrid material combines the advantages of an organic biopolymer, Alg and inorganic LDHs. Glucose oxidase (GOD) immobilized in the material maintained its activity. The composite films were characterized by UV-Vis. The results indicated that GOD retained the essential feature of its native structure in the composite film. The Alg/LDHs/GOD-modified platinum electrode exhibited a fast response to glucose (achieve 95% of the maximum in 10 s), which provided a linear response to glucose over a concentration range of 1.6 x 10(-5)-2 x 10(-3) M with a detection limit of 4 x 10(-5) M based on S/N = 3. Furthermore, the biosensor exhibited excellent long-term stability, and satisfactory reproducibility. It retained 87% of its original activity after being used for 28 days. PMID- 20009329 TI - Determination of chromium(VI) in electronics materials using trioctylamine modified carbon paste electrode. AB - A trioctylamine (TOA) modified carbon paste electrode (TOA/CPE) was firstly utilized to determine Cr(VI) in electronics materials. The effects of preconcentration conditions, that is, TOA amount and accumulation time on Cr(VI) accumulation were examined and the optimum experiment conditions for the determination were identified. A sensitive reduction peak in the stripping voltammogram at -0.45 V, a characteristic of trace Cr(VI), was detected when the accumulation time was 10 min. Under optimized conditions, TOA/CPE demonstrated an enhanced sensitivity for Cr(VI), providing a low detection limit (S/N = 3) at 3.4 x 10(-9) M. Interference studies also displayed high selectivity of the TOA/CPE for Cr(VI); this electrode can accurately determine Cr(VI) in the presence of Cr(III) (600-fold concentration) and other interfering cations. PMID- 20009330 TI - Carbon paste- and PVC membrane electrodes as sensitive sensors for the determination of antidiabetic drugs for type 2 diabetic patients. AB - Carbon paste- and polyvinyl chloride membrane electrodes are simple, precise, rapid and selective sensors for the determination of antidiabetic drugs for type 2 diabetic patients. These electrodes were successfully used for the potentiometric determination of rosiglitazone, pioglitazone, glimepiride and glyburide in their standard forms and also as pharmaceutical preparations. The preparation of these ion-selective electrodes for the potentiometric determination of the drug is based on the construction of a 10% standard drug-ion pair with reineckate or tungstophosphate imbedded as an electro-active material in the carbon paste or in the polyvinyl chloride membrane. The prepared ion selective electrodes showed a Nernstian response with a limit of detection amounting to 10(-6) M in a pH range of 3 to 5. A good selectivity coefficient and long term stability could be achieved. The developed potentiometric method based on the CPE and PVC sensors is economic and less time consuming compared to the conventionally used high performance liquid chromatography, HPLC, methods. PMID- 20009331 TI - Determination of fluoride using ion-selective electrodes in the presence of aluminum. AB - We describe a method for determining fluoride with ion-selective electrodes (ISEs). Tartrate and Tris-based total ionic strength adjustment buffers (TISABs) were found to lower the interference from aluminum to a greater extent than conventional citrate-based TISABs. We adopted a solid TISAB addition method that is simple to perform, and can be carried out without lowering the level of fluoride. The apparent recovery of fluoride was 95% or higher, even at 500 mg L( 1) of Al3+ when a tartrate and Tris-based TISAB was used. Interferences from common ions were not observed at 100 mg L(-1) levels. We determined the fluoride content in solid silicate samples with ISEs without preliminary steam distillation after alkali fusion processing. Adding a solid TISAB mixture consisting of tartaric acid, sodium tartrate, and Tris, however, eliminated any interference from high levels of aluminum and sodium and potassium carbonates. The proposed analytical method was also applied to the determination of fluoride in geochemical reference samples. PMID- 20009332 TI - Improved solid-phase spectrophotometry for the microdetermination of chromium(VI) in natural water. AB - A simple and sensitive solid-phase spectrophotometry procedure was improved for the microdetermination of Cr(VI). A 0.06 cm3 portion of a cation exchanger, Muromac AG 50W-X2, was used to concentrate the target Cr(VI) in a 20 cm3 water sample, and resin beads were introduced in a flow cell of 1.5 mm diameter and having a 10 mm light path length for measurements using a UV-visible spectrophotometer. Three lenses were used for focusing the incident light beam and for recovering light scattered by the solid phase in the cell. The sensitivity achieved was higher by a factor of 277 compared with that of the solution method, and the detection limit was 0.014 microg dm(-3). The recovery on spiked real water samples by the standard addition method was 96-101%. Favorable working and performance characteristics made it possible to directly determine sub-microg dm(-3) amounts of Cr(VI) in natural water samples. PMID- 20009333 TI - Spectrophotometric determination of dopamine hydrochloride in pharmaceutical, banana, urine and serum samples by potassium ferricyanide-Fe(III). AB - In the present work, we developed a simple, sensitive and inexpensive method to determine dopamine hydrochloride using potassium ferricyanide-Fe(III) by spectrophotometry. The results show that Fe(III) is deoxidized to Fe(II) by dopamine hydrochloride at pH 4.0, and then Fe(II) reacts with potassium ferricyanide to form a soluble prussian blue (KFe(III)[Fe(II)(CN)6]). The absorbance of this product was monitored over time using a spectrophotometer at an absorption maximum of 735 nm, and the amount of dopamine hydrochloride could be calculated based on the absorbance. A good linear relationship of the concentration of dopamine hydrochloride versus absorbance was observed, and a linear regression equation of A = 0.022 + 0.16921C (microg mL(-1)) was obtained. Moreover, the apparent molar absorption coefficient for the indirect determination of dopamine hydrochloride was 3.2 x 10(4) L mol(-1) cm(-1). This described method has been used to determine dopamine hydrochloride in pharmaceutical, banana, urine and serum samples with satisfactory results. PMID- 20009334 TI - Spectrophotometric determination of copper(II) with o-bromophenylfluorone. AB - A simple, reliable and sensitive spectrophotometric method for the determination of copper(II) was established with o-bromophenylfluorone (OBPF), a novel chemical probe, in the presence of poly(N-vinylpyrrolidone). Beer's law was obeyed in the range of 8-160 ng ml(-1), with an apparent molar absorptivity at 570 nm, the relative standard deviation being 3.64 x 10(5) dm3 mol(-1) cm(-1) and 0.72% (n = 5). This method was applied to the recovery tests of copper(II) in human urine, bovine serum and tap water; the results were satisfactory. PMID- 20009335 TI - Fast determination of the release degree of compound sulfamethoxazole tablets by using circulation release system combined with ultraviolet spectral net analyte signal method. AB - Based on ultraviolet spectral net analyte signals, this research has studied the determination of the two effective components, sulfamethoxazole (SMZ) and trimethoprim (TMP), contained in compound sulfamethoxazole tablets in acid environment. The linear ranges of SMZ and TMP were 0.48-7.84 microg/ml (the regression correlation coefficient r = 0.9981) and 0.12-1.5 microg/ml (r = 0.9986), the corresponding average recoveries were 99.5 and 101.0%, respectively, and the relative standard deviations were 1.87 and 3.60%, respectively. The method was simple, fast and accurate. A circulation release system for the determination of the tablet release degree has been built, and the on-line filtering and small-volume sampling could thus be carried out. Thanks to the combination of the circulation release system and the determination method of SMZ and TMP in acid environment, the determination of release degree for compound sulfamethoxazole tablets was fast and accurate, especially for the early time of rapid release. PMID- 20009336 TI - Determination of nitric oxide using horseradish peroxidase by UV second-order derivative spectrometry. AB - Based on the reaction of horseradish peroxidase (HRP) with nitric oxide (NO), a novel detection method of NO has been developed. The method uses second-order derivative spectrometry in an anaerobic phosphate buffer solution. The effects of pH and HRP concentration on the determination of NO in HRP system were investigated, and the conditions for the measurements were optimized. Some possible coexisting substances, such as nitrite, nitrate and hydrogen peroxide, were tested. The linear regression equation of standard curve was found to be h = 8.89 x 10(-2)c(NO)-1.56 x 10(-3) with the relevant coefficient of 0.9966 (n = 5) in the NO concentration range of 0.085-1.3 microM. The relative standard deviations were less than 3%. Based on the standard deviation of 5 blank measurements and a signal-to-noise ratio of 3, the detection limit for NO was 0.032 microM. The proposed method was successfully applied to the determination of NO levels in serum samples. PMID- 20009337 TI - Determination of Hg and Pb in fuels by inductively coupled plasma mass spectrometry using flow injection chemical vapor generation. AB - An isotope dilution inductively coupled plasma mass spectrometry (ICP-MS) method has been developed for the determination of Hg and Pb in fuels using flow injection vapor generation (VG) as the sample introduction system. A simple and inexpensive in-situ nebulizer/vapor generator was employed in this study. An emulsion containing 10% v/v fuel, 2% m/v Triton X-100 and 1.0% m/v tartaric acid was injected into VG-ICP-MS system for the determination of Hg and Pb. Sodium borohydride was used for vapor generation. Since the sensitivities of Hg and Pb in emulsion and those in aqueous solution are quite different, isotope dilution and standard addition methods were used for the determination of Hg and Pb in selected fuel samples. The influences of vapor generation conditions and emulsion preparation on the ion signals are reported. This method has been applied for the determination of Hg and Pb in various fuel samples such as diesel, gasoline and engine oil obtained locally. The analytical results obtained by isotope dilution and standard addition methods were in good agreement with each other and also with those of digested samples analyzed by pneumatic nebulization ICP-MS. Under the optimum operating conditions, the detection limits obtained were 0.02 and 0.03 ng mL(-1) for Hg and Pb, respectively, in prepared emulsified solutions, corresponding to 0.2 and 0.3 ng mL(-1) of Hg and Pb, respectively, in the original fuel samples. PMID- 20009338 TI - Silica gel chemically modified with ethyl-2-benzothiazolylacetate for selective extraction of Pb(II) and Cu(II) from real water samples. AB - A silica-gel chemically modified with ethyl-2-benzothiazolylacetate (SiB) has been synthesized and characterized. The extractions towards Pb(II), Cu(II), Co(II), Cd(II) and Ni(II) were carried out in batch extraction. The maximum extraction was found at pH > 4 within 10 min. The maximum adsorption capacities, evaluated from Langmuir's model were 221, 141, 13, 7 and 5 micromol g(-1) for Pb(II), Cu(II), Co(II), Cd(II) and Ni(II), respectively. Home-made mini-columns were used in the flow system. The parameters that affected the extraction efficiency were explored. Sample volumes of 100.0 mL and 1% HNO3 of 5.0 mL as the eluent were employed under the optimized conditions. No interfering from Na+, K+, Mg2+, Ca2+, Cl- and SO(4)(2-) was observed. This modified silica gel was applied to preconcentrate Pb(II) and Cu(II) from spiked pond water, tap water and drinking water, giving high accuracy and precision. The method detection limits obtained from the desorbed metals determined by FAAS after preconcentration with SiB were 22 and 1 microg L(-1) for Pb(II) and Cu(II), respectively. PMID- 20009339 TI - Terahertz absorption spectra of original and generic ceftazidime. AB - The absorption spectra of ceftazidime and its generic versions (Modacin, Mosyl, and Mobenzocin) have been measured by terahertz time domain spectroscopy (THz TDS). Differences in the THz absorption were observed between the original and generic versions. The results show small, but significant differences in the states of ceftazidime hydrate between the original and generic versions. THz-TDS can be used to evaluate the stability of medicines as well as to control their quality. PMID- 20009340 TI - Single-tube genotyping from a human hair root by direct PCR. AB - We have developed a simple, labor-saving, inexpensive and rapid SNP genotyping method that directly uses a human hair root as the template. This single-tube genotyping method was used to successfully and reliably genotype the ADH1B and ALDH2 polymorphisms using a hair root (without DNA isolation) and the polymerase chain reaction (PCR) enzyme kit KOD FX. Since the DNA extraction step was eliminated, the possibility of sample contamination was considerably decreased. The single-tube SNP genotyping was performed by coupling the PCR enzyme kit with allele-specific primer (ASP)-PCR. In the post-genome era, a simple and inexpensive method for diagnostic analysis is in high demand, and this PCR method with a hair root as a template will be very useful for genetic diagnoses in biological and medical laboratories. PMID- 20009341 TI - Determination of chloroacetic acids in drinking water using suppressed ion chromatography with solid-phase extraction. AB - Suppressed ion chromatography with a conductivity detector was developed for the determination of trace amounts of underivatized chloroacetic acids (CAAs). When sodium carbonate and methanol were used as a mobile phase, the simultaneous determination of each CAA took approximately 25 min. The linearity, reproducibility and detection limits were determined for the proposed method. For the solid-phase extraction step, the effects of the pH of the sample solution, sample volume and the eluting agent were tested. Under the optimized extracting conditions, the average recoveries for CAAs spiked in tap water were 83-107%, with an optimal preconcentration factor of 20. The reproducibility of recovery rate for CAAs was 1.2-3.8%, based upon 6 repetitions of the recovery experiments. PMID- 20009342 TI - Simultaneous determination of amlodipine besylate and benazepril hydrochloride in pharmaceutical dosage form by LC. AB - A rapid, precise, specific, and accurate ultra performance liquid chromatography (UPLC) method has been developed and subsequently validated for simultaneous determination of amlodipine present as amlodipine basylate (AB) and benazepril hydrochloride (BH) in capsule dosage form. The chromatographic separation was achieved on an Acquity UPLC, BEH C8 (100 mm x 2.1 mm, 1.7 microm) column using a photodiode array detector. The mobile phase used consisted of a mixture of phosphate buffer pH 3.0 (0.01 M aqueous potassium dihydrogen phosphate, pH 3.0 adjusted with orthophosphoric acid) and solvent mixture (equal mixture of acetonitrile and methanol) in the ratio of 45:55 (v/v) at a flow rate of 0.3 mL/min. The described method was linear over ranges of 5.21-15.63 microg/mL for AB and 20.24-60.72 microg/mL for BH. The mean recoveries were 100.47 and 99.97% for AB and BH, respectively. The lower limit of quantification was determined on the basis of signal-to-noise ratio method; it is 0.01 microg/mL for AB and 0.019 microg/mL for BH. PMID- 20009344 TI - Simultaneous immunohistochemical detection of gangliosides and neuronal markers in paraformaldehyde-fixed nervous tissues by acetone etching. AB - A need for identifying ganglioside-positive cells with neuronal markers prompted us to establish a reliable method for double or triple immunostaining nervous tissues. Perfusion fixation with paraformaldehyde is typically performed for the routine immunostaining of various neuronal markers but is not suitable for immunostaining gangliosides. Acetone fixation of fresh cryosections is frequently used for ganglioside immunodetection; thus, we tested the effect of acetone treatment for unmasking the antigen epitope of gangliosides (acetone etching) on sections of paraformaldehyde-fixed nervous tissue from rats. Acetone etching significantly retrieved ganglioside immunoreactivity while preserving the immunoreactivity of neuronal markers. Various combinations of gangliosides and neuronal markers could be double-stained by the immunoenzyme method or triple stained by the immunofluorescence method. This new method may provide additional information regarding the relationship between gangliosides and various neuronal markers from routinely paraformaldehyde-fixed nervous tissues, both freshly prepared specimens and those stocked in the laboratory. PMID- 20009345 TI - Histochemical changes and apoptosis in degenerating taste buds of the rat circumvallate papilla. AB - The present study was designed to examine the histochemical changes and occurrence of apoptosis in taste buds of rat circumvallate papillae following bilateral transection of the glossopharyngeal nerve. Following transection of the glossopharyngeal nerve, the number of taste buds was not altered until post operative day 3 (PO3), but decreased significantly thereafter. The number of cells within a taste bud, however, decreased significantly from PO2. In normal, uninjured animals, approximately 15.4%, 9.0%, and 7.7% of taste bud cells were labeled with antibodies for phospholipase C beta2 subunit (PLCbeta2), a marker for type II cells, neural cell adhesion molecule (NCAM), a marker for type III cells, and Jacalin, a marker for type IV cells, respectively. Following gustatory nerve injury, the ratio of cells expressing markers of type III and type IV decreased gradually from PO2, and Jacalin-labeled taste bud cells disappeared on PO3. Under normal conditions, immunoreactivity for single-strand DNA (ssDNA), a marker of apoptosis, was detected in the nuclei of PLC beta2-immunoreactive cells and cells showing no labeling for PLCbeta2, NCAM, or Jacalin. On PO1, the number of taste bud cells showing ssDNA immunoreactivity increased to double that of normal uninjured animals; these ssDNA-immunoreactive cells were also labeled with NCAM and Jacalin as well as PLCbeta2. The present results suggest that denervation of the gustatory nerve causes apoptosis in all types of taste bud cells, resulting in the rapid degeneration of taste buds. PMID- 20009346 TI - Low vacuum scanning electron microscopy for paraffin sections utilizing the differential stainability of cells and tissues with platinum blue. AB - The present study introduces a novel method for the direct observation of histological paraffin sections by low vacuum scanning electron microscopy (LVSEM) with platinum blue (Pt-blue) treatment. Pt-blue was applied not only as a backscattered electron (BSE) signal enhancer but also as a histologically specific stain. In this method, paraffin sections of the rat tongue prepared for conventional light microscopy (LM) were stained on glass slides with a Pt-blue staining solution (pH 9) and observed in a LVSEM using BSE detector. Under LVSEM, overviews of whole sections as well as three-dimensional detailed observations of individual cells and tissues could be easily made at magnifications from x40 to x10,000. Each kind of cell and tissue observed in the section could be clearly distinguished due to the different yields of BSE signals, which depended on the surface structures and different affinities to Pt-blue. Thus, we roughly classified cellular and tissue components into three groups according to the staining intensity of Pt-blue observed by LM and LVSEM: 1) a strongly stained (deep blue by LM and brightest by LVSEM) group which included epithelial tissue, endothelium and mast cells; 2) a moderately stained (light blue and bright) group which included muscular tissue and nervous tissue; 3) an unstained or weakly stained (colorless and dark) group which included elastic fibers and collagen fibers. We expect that this method will prove useful for the three-dimensional direct observation of histological paraffin sections of various tissues by LVSEM with higher resolutions than LM. PMID- 20009347 TI - Immunohistochemical demonstration of c-Kit-negative fibroblast-like cells in murine gastrointestinal musculature. AB - In the gastrointestinal musculature, interstitial cells of Cajal (ICC) distribute and regulate the gastrointestinal motility. Another type of mesenchymal cell, known as the fibroblast-like cell (FLC), has also been reported to be juxtaposed to the ICC. In this study, we examined the immunohistochemical properties of FLC in the murine gastrointestinal musculature using antibodies to small conductance Ca(2+)-activated K(+) channel 3 (SK3), platelet-derived growth factor receptor alpha (PDGFRalpha), and CD34. SK3-immunopositive (SK3-ip) cells were observed in the musculature throughout the gastrointestinal tract. These SK3-ip cells were distinct from the ICC that were identified by c-Kit immunoreactivity. In the muscular layers, SK3-ip cells were bipolar in shape and were associated with the intramuscular ICC and nerve fiber bundles. In the myenteric layer multipolar shaped SK3-ip cells encompassed the myenteric ganglia. SK3-ip cells in the subserosal plane formed a cellular network with their ramified processes. The distribution pattern of the SK3-ip cells in the ICC-deficient W(v)/W(v) mutant mice was similar to that in normal mice. We also demonstrated that SK3-ip cells showed the intense PDGFRalpha immunoreactivity that was previously examined in FLC. However, CD34 immunoreactivity, one of the markers of human FLC, was not observed in SK3-ip cells with the exception of subserosal FLC. Thus, our observations indicate that SK3- and PDGFRalpha-double immunopositive cells are FLC in the murine gastrointestinal musculature and behave as a basic cellular element throughout the gastrointestinal musculature. PMID- 20009348 TI - The expression of soluble guanylate cyclase in the vasculature of rat skeletal muscle. AB - Nitric oxide (NO) has various roles in the skeletal musculature in both normal and pathological conditions. NO primarily activates soluble guanylate cyclase (sGC) and mediates subsequent intracellular signaling in target cells. We sought to identify the target cells of NO in the rat skeletal musculature, using subtypes of sGCalpha1 and sGCbeta1 antibodies. Immunohistochemistry revealed that both antibodies stained the same cells with round or oval shapes, having several long processes. The sGC-immunopositive cells co-expressed NG2 chondroitin sulfate proteoglycan, a marker of pericytes. The sGC-immunopositive cells were associated with capillaries and formed cellular networks with elongated cytoplasmic processes. sGCalpha1 and sGCbeta1 were not found in muscle sarcolemma that were stained by anti-dystrophin, or neuromuscular junctions, as detected by anti synaptophysin. Based on these findings, we concluded that sGC immunoreactivity was specifically distributed in capillary pericytes. Pericytes in the skeletal musculature have been shown to be target cells of NO and are involved in the microvascular blood flow. PMID- 20009349 TI - Transplanted embryonic spinal tissue promotes severed sciatic nerve regeneration in rats. AB - The effects of transplanted embryonic spinal tissue on host motor nerve regeneration and target muscle reinervation were investigated in severed sciatic nerves of rats. The electromyogram (EMG) responses and number of motor end plates (MEP) in target muscles, number of nerve axons, and retrogradely labeled motor neurons were examined in transplantation-, anastomosis without transplantation-, and naive groups of the animals. The EMG patterns of the transplantation group returned to nearly normal at the 8th week, but those of the anastomosis group did not. MEP counts in the transplantation group were significantly higher than in the anastomosis group. The myelinated axon counts and myelin sheath thickness in the transplantation group were significantly higher than those in the anastomosis group. The number of retrogradely labeled motor neurons was significantly higher in the transplantation group. We conclude that transplanted embryonic spinal tissue can promote both host motor nerve regeneration and target muscle reinnervation. PMID- 20009350 TI - Evaluation of the hemodynamic impact of continuous renal replacement therapy in healthy dogs. AB - We performed continuous renal replacement therapy (CRRT) on clinically healthy dogs to evaluate the effects of CRRT on hemodynamics. Heart rate, arterial blood pressure, and central venous pressure of the dogs (n=6) were recorded during the procedure, which was performed under general anesthesia. Throughout the CRRT, heart rate and arterial blood pressure were stable. Central venous pressure increased after CRRT termination but returned to the basal level within 30 min. In this study, hemodynamic alterations, including hypotension, hypertension, and arrhythmias, were not observed during CRRT. These observations suggest that the CRRT protocol used in the present study can be safely applied to clinical cases with acute renal failure. PMID- 20009351 TI - Effects of several pyrethroids on hepatic cytochrome P450 activities in rats. AB - Four commonly used pyrethroids (permethrin, bifenthrin, ethofenprox, and fenpropathrin) were orally administered to Sprague-Dawley rats for 5 days to study their effects on the liver cytochrome P450 (CYP) activities. Also Michaelis Menten kinetics of the metabolic reactions catalyzed by liver CYPs were examined after adding these pyrethroids to the assay system to investigate their possible inhibitory effects on liver CYPs activities. These reactions included ethoxyresorufin O-deethylation, tolbutamide hydroxylation, bufuralol 1' hydroxylation, and midazolam 4-hydroxylation, for CYP1A, 2C, 2D, and 3A activities, respectively. Results showed that oral administration of bifenthrin and ethofenprox highly induced CYP1A. The most potent inhibitors for CYP1A were fenpropathrin and cis-permethrin with K(i) values of 3.71 & 3.87 microM, respectively. CYP2D was slightly inhibited by both of fenpropathrin and cis permethrin (K(i) values were 307.32 & 632.23 microM, respectively). On the other hand, none of CYP2C or 3A was inhibited by the tested pyrethroids. Since CYP1A may relate to biotransformation of many chemicals to reactive metabolites, bifenthrin and ethofenprox may potentiate mutagenicity of the chemicals through their inducing effects on CYP 1A. As permethrin and fenpropathrin were potent inhibitor for CYP1A, they may result in substantial accumulation of some chemicals. The resultant accumulation may lead to fatal toxicities in some case. PMID- 20009352 TI - Isolation of Trypanosoma (Megatrypanum) theileri from dairy cattle in Taiwan. AB - Although Trypanosoma (Megatrypanum) theileri, a blood parasite of bovid species, is spread widely throughout the world, it has never been reported in Taiwan. When an anti-coagulated blood sample from febrile dairy cattle was directly smeared, no parasite was observed. However, a highly distinctive morphological feature of trypanosome appeared in baby hamster kidney (BHK) cell culture inoculated with non-thrown blood buffy coat. The different stages and typical ultrastructures of trypanosome were observed in our isolate. The isolate was subsequently identified as T. theileri by species-specific PCR assay (Tth625), 18S rDNA sequencing alignment and internal transcribed spacer of ribosomal genes (ITS) as a marker for molecular phylogenetic analysis. The first T. theileri isolate in Taiwan (TWTth1) could be periodically passaged in BHK cell culture for more than one year and retained good re-cryopreservation viability. The BHK culture method would be excellent for diagnostic isolation and maintenance long-term development of this parasite. PMID- 20009353 TI - The mode of biofilm formation on smooth surfaces by Campylobacter jejuni. AB - Many microorganisms produce extracellular polymers referred to collectively as "slime" or glycocalyx, and form biofilms on solid surfaces in natural ecosystems. Campylobacter jejuni, one of the most important foodborne pathogens, also has the ability to form biofilm on stainless steel, glass, or polyvinyl chloride in vitro. However, the issue of biofilm formation by Campylobacter species has not been extensively examined. The present study was performed to examine the mode of adhesion of C. jejuni to a smooth surface. When bacterial suspensions in Brucella broth were incubated in microplate wells with a glass coverslip, microcolonies 0.5~2 mm in diameter were formed on the coverslip within 2 hr from the start of incubation. These microcolonies gradually grew and formed a biofilm of net-like connections within 6 hr. Transmission electron microscopy indicated that massive amounts of extracellular material masked the cell surface, and this material bound ruthenium red, suggesting the presence of a polysaccharide moiety. Scanning electron microscopy indicated that the flagella acted as bridges, forming net like connections between the organisms. To determine the genes associated with biofilm formation, aflagellate (flaA(-)) and flagellate but non-motile (motA(-)) mutants were constructed from strain 81-176 by natural transformation-mediated allelic exchange. The flaA(-) and motA(-) mutants did not form the biofilm exhibited by the wild-type strain. These findings suggest that flagella-mediated motility as well as flagella is required for biofilm formation in vitro. PMID- 20009354 TI - Characteristics of chronic rejection in heart transplantation: important elements of pathogenesis and future treatments. AB - Although 85,000 heart transplantations have been performed worldwide, coronary allograft vasculopathy (CAV), which is a phenomenon of chronic rejection, is still a serious problem. Because CAV involves all the allograft arteries, angioplasty, stenting or bypass grafting are not practical treatment options. Therefore, CAV is the biggest long-term limitation in cardiac allograft recipients. Although the cause of CAV is mostly immunologic, nonimmune pathways also contribute to its development. Several cytokines, chemokines and adhesion molecules play a critical role in the process. Cell adhesion, migration and proliferation of bone marrow progenitor and and other cells are involved in its development. Although there is not an established clinical strategy for preventing or treating CAV, recent investigations have provided some promising methodologies. Progress in DNA technology, such as antisense oligodeoxynucleotides (ODNs) to regulate the transcription of disease-related genes, has an important role in its therapeutic applications. Antisense ODN transfection preventing CAV in experimental cardiac allografts has been reported for the first time. The ODN strategy has not only been useful in the experimental studies, but is also a novel clinical strategy for gene therapy. The pathological and immunological characteristics of CAV and some promising methodologies for prevention of the disease are reviewed. PMID- 20009355 TI - Long-term prognosis of isolated significant tricuspid regurgitation. AB - BACKGROUND: The prognostic implications and surgical benefit of isolated significant tricuspid regurgitation (TR) and prognostic factors in patients with TR were investigated. METHODS AND RESULTS: In 870 consecutive patients with significant isolated TR, all-cause mortality was analyzed over 4.9+/-2.9 years. It was found that the survival rate tended to be higher in the 57 patients who underwent tricuspid valve (TV) surgery than the 813 patients who did not by using propensity-score matching (P=0.068), although it was not significant. Of the 813 patients that did not undergo TV surgery, the 5-year survival rate was 74%. According to the Cox proportional hazards model, the initial TR jet area (hazard ratio [HR], 1.044; 95% confidence interval [CI], 1.016-1.073), pulmonary artery systolic pressure (HR, 1.024; 95%CI, 1.017-1.032) and presence of right ventricular (RV) dysfunction (HR, 2.256; 95% CI, 1.329-3.828) were predictors of mortality independent of patient age and presence of diabetes mellitus and renal failure in medically managed patients. CONCLUSIONS: In patients with isolated significant TR, there was a tendency, although not significant, towards a higher survival rate after TV surgery. The severity of TR and pulmonary hypertension, and the presence of RV dysfunction are independent prognostic factors in medically managed patients. Further prospective randomized studies are necessary to demonstrate the benefit of TV surgery in these poor prognostic populations. PMID- 20009356 TI - Relationship between multiple plasma biomarkers and vulnerable plaque determined by virtual histology intravascular ultrasound. AB - BACKGROUND: The relationship between plasma biomarkers and vulnerable plaque is not well understood. METHODS AND RESULTS: The 188 patients who underwent 3-vessel virtual histology (VH) intravascular ultrasound (IVUS) with peripheral blood sampling were enrolled. Plasma levels of matrix metalloproteinase 2 and 9 (MMP-2, -9), tissue inhibitor of metalloproteinase-1, adiponectin, and macrophage migration inhibitory factor were measured. VH-IVUS-derived thin cap fibroatheroma (VH-TCFA) was defined as a necrotic core >10% of plaque area in the presence of >40% plaque burden. There were 38 patients with ruptured plaque and 150 patients without (107 patients with VH-TCFA, 43 patients without VH-TCFA) in culprit/target lesions. Among the biomarkers, only the MMP-9 level was significantly higher in patients with ruptured plaque (P=0.002). In the subgroup without ruptured plaque, significant differences in the levels of several biomarkers were not observed between patients with and without VH-TCFA. In both culprit/target and nonculprit/non-target vessels, the MMP-9 level showed a weak correlation with the total number of ruptured plaques (r=0.231, P=0.002). CONCLUSIONS: Among the biomarkers tested in this study, the MMP-9 level was significantly higher in patients with ruptured plaque. However, measurement of several biomarkers, including MMP-9, was incapable of predicting the presence of VH-TCFA. PMID- 20009357 TI - Fasting serum level of fatty-acid-binding protein 4 positively correlates with metabolic syndrome in patients with coronary artery disease. AB - BACKGROUND: The serum level of fatty-acid-binding protein 4 (FABP4) increases in patients with metabolic syndrome (MetS), so the relationship between FABP4 and MetS among patients with coronary artery disease (CAD) was investigated in the present study. METHODS AND RESULTS: Fasting blood samples were obtained from 98 CAD patients. MetS and its components were defined using the diagnostic criteria of the International Diabetes Federation; 50 CAD patients (51.0%) had MetS. The fasting level of FABP4 positively correlated with MetS (P=0.037) and serum levels of FABP4 correlated with a number of MetS criteria (P=0.035). Univariate linear regression analysis showed that body fat mass (R=0.234; P=0.020) and the levels of triglycerides (R=0.348; P<0.001), and low-density lipoprotein-cholesterol (R=0.217; P=0.032) positively correlated with the serum level of FABP4, whereas the level of high-density lipoprotein-cholesterol (R=-0.243; P=0.016) negatively correlated with it. Multivariate forward stepwise linear regression analysis of the significant variables showed that the level of triglycerides (beta=0.348, R(2)=0.121, P<0.001) was the independent predictor of fasting serum level of FABP4. CONCLUSIONS: Among CAD patients in the present study, the fasting level of FABP4 positively correlated with MetS and serum levels of FABP4 correlated with a number of MetS criteria. PMID- 20009358 TI - Novel ECG predictor of difficult cases of outflow tract ventricular tachycardia: peak deflection index on an inferior lead. AB - BACKGROUND: An ECG predictor of ablation success has not been determined for difficult cases of outflow tract ventricular tachycardia/ventricular premature contractions (OT-VT/VPC). METHODS AND RESULTS: ECG analysis and radiofrequency catheter ablation (RFCA) were performed in 70 patients with OT-VT/VPC. The peak deflection index (PDI) was determined in the inferior lead presenting the tallest R wave by dividing the time from QRS onset to peak QRS deflection by total QRS duration. In 10 (14%) of the 70 patients, RFCA performed at a septal or epicardial site was unsuccessful (group 1), but was successful in the remaining 60 patients (group 2). Neither activation time (35+/-15 ms vs 40+/-12 ms, P=0.3) nor QRS duration (141+/-19 ms vs 137+/-19 ms, P=0.6) were significantly different between groups 1 and 2. However, PDI was significantly higher in group 1 than in group 2 (0.62+/-0.06 vs 0.55+/-0.06, P=0.002). A PDI >0.6 identified unsuccessful OT-VT/VPC with 80% sensitivity and 90% specificity, and may indicate that the origin of the OT-VT/VPC is deep within the ventricular septum or at an epicardial site. CONCLUSIONS: A PDI >0.6 is more likely to be associated with a higher rate of RFCA failure. PMID- 20009359 TI - Expression of toll-like receptors in the pancreas of recent-onset fulminant type 1 diabetes. AB - Fulminant type 1 diabetes, established in 2000, is defined as a novel subtype of diabetes mellitus that results from remarkably acute and almost complete destruction of pancreatic beta cells at the disease onset. In this study, we aimed to clarify the pathogenesis of fulminant type 1 diabetes with special reference to insulitis and viral infection. We examined pancreatic autopsy samples from three patients who had died soon after the onset of disease and analyzed these by immunohistochemistry and in situ-hybridization. The results were that both beta and alpha cell areas were significantly decreased in comparison with those of normal controls. Mean beta cell area of the patients just after the onset was only 0.00256 % while that of normal control was 1.745 %. Macrophages and T cells-but no natural killer cells-had infiltrated the islets and the exocrine pancreas. Although both of them had massively infiltrated, macrophages dominated islet infiltration and were detected in 92.6 % of the patients' islets. Toll-like receptor (TLR) 3, a sensor of viral components, was detected in 84.7+/- 7.0 % of T cells and 62.7+/- 32.3 % of macrophages (mean+/- SD) in all three patients. TLR7 and TLR9 were also detected in the pancreas of all three patients. Enterovirus RNA was detected in beta-cell positive islets in one of the three patients by in situ-hybridization. In conclusion, our results suggest that macrophage-dominated insulitis rather than T cell autoimmunity contributes to beta cell destruction in fulminant type 1 diabetes. PMID- 20009360 TI - Tumor necrosis factor-alpha (TNF-alpha) inhibits insulin-like growth factor-I (IGF-I) activities in human trophoblast cell cultures through IGF-I/insulin hybrid receptors. AB - Tumor necrosis factor-alpha (TNF-alpha) in placenta is believed to be involved in pathogenesis of intrauterine growth restriction. In contrast, insulin-like growth factors (IGFs) are believed to be important for stimulation of fetal and placental growth. IGF-I stimulates metabolic and growth-promoting actions directly through its receptors: IGF-I receptor (IGF-IR), insulin receptor (IR) and IGF-I/insulin hybrid receptor (HR). However, it has not been elucidated which receptor mediates the growth promoting effects in fetal and placental growth. The current studies were undertaken to test whether TNF-alpha affects IGF-I action on placenta using human trophoblast cell cultures, and to test which receptor mediates growth promoting effects of IGF-I in placenta. Primary culture of trophoblast cells, which express IGF-IR, IR, and HR, were exposed to TNF-alpha, and the effects of IGF-I in stimulating trophoblast cell proliferation and migration were determined. Exposure to TNF-alpha attenuated the effects of IGF-I on cell proliferation and migration. To determine which receptors are involved in this inhibitory effect, the ability of IGF-I to stimulate phosphorylation of its receptors was analyzed in the presence of TNF-alpha. TNF-alpha exposure neither attenuated the phosphorylation of IGF-IR homodimer by IGF-I nor changed receptor abundance. In contrast, TNF-alpha reduced the ability of IGF-I to stimulate phosphorylation of HR with reducing amounts of HR. Exposure to TNF-alpha also attenuated phosphorylation of insulin receptor substrate-1 (IRS-1) and the association of IRS-1 with phosphatydilinositol-3 kinase (PI-3 kinase). Taken together, these findings indicate that TNF-alpha induces a loss of sensitivity to stimulation by IGF-I, through reducing amounts of HR and the stimulation of HR tyrosine kinase activity by IGF-I. PMID- 20009361 TI - [Pain relief by gabapentin via supraspinal mechanisms in neuropathic conditions]. PMID- 20009362 TI - [Depression and corticosteroid receptors]. PMID- 20009363 TI - [Current therapeutic approaches and involvement of amyloid-beta in retinal diseases]. PMID- 20009364 TI - [Utilization of remnant human specimens provided by the human tissue bank for biomedical research]. PMID- 20009365 TI - [Direct measurement of the QT interval in stem cell-derived cardiomyocytes for the assessment of QT liability]. PMID- 20009366 TI - [A novel experimental model for benign prostatic hyperplasia: stromal hyperplasia model in rats]. PMID- 20009367 TI - [CYP induction screening using cryopreserved human hepatocytes in drug development]. PMID- 20009368 TI - [Evaluation of phase II drug metabolizing enzymes in the discovery stage]. PMID- 20009369 TI - [Evaluation methods for reactive metabolites]. PMID- 20009370 TI - Sleep duration and incidence of cardiovascular events in a Japanese population: the Jichi Medical School cohort study. AB - BACKGROUND: Although sleep is one of the most important health-related factors, the relationship between sleep duration and the incidence of cardiovascular events has not been fully described. METHODS: The present study comprised the 11,367 study subjects (4413 men and 6954 women) of the Jichi Medical School Cohort Study, a population-based prospective study. Baseline data were obtained by questionnaire and health examinations between April 1992 and July 1995 in 12 rural areas in Japan, and the main outcome measures were the incidence of cardiovascular diseases (stroke and myocardial infarction [MI]). Cox proportional hazards models were used to analyze the association between sleep duration and the incidence of cardiovascular events. RESULTS: A total of 481carciovascular events (255 men and 226 women) were observed during an average follow-up period of 10.7 years. After adjusting for age, systolic blood pressure, serum total cholesterol, body mass index, smoking habits, and alcohol drinking habits, the hazard ratios (95% confidence intervals) for the incidence of cardiovascular diseases for individuals sleeping less than 6 hours and 9 hours or longer were 2.14 (1.11-4.13) and 1.33 (0.93-1.92) in men, and 1.46 (0.70-3.04) and 1.28 (0.88 1.87) in women, respectively, relative to those who reported sleeping 7 to 7.9 hours per day. CONCLUSIONS: Our data indicate that men who sleep less than 6 hours a day have a higher risk of cardiovascular events than those sleeping 7 to 7.9 hours. PMID- 20009372 TI - Combined factors effect of menstrual cycle and background noise on visual inspection task performance: a simulation-based task. AB - It is well known that women are physiologically and psychologically influenced by the menstrual cycle. In addition, the presence of background noise may affect task performance. So far, it has proven difficult to describe how the menstrual cycle and background noise affect task performance; some researchers have found an increment of performance during menstruation or during the presence of noise, others found performance deterioration, while other still have reported no dominant effect either of the menstrual cycle in performance or of the presence of noise. However, no study to date has investigated the combinational effect between the menstrual cycle and the presence of background noise in task performance. Therefore, the purpose of this study was to examine the combined factor effect of menstrual cycle and background noise on visual inspection task performance indices by Signal Detection Theory (SDT) metrics: sensitivity index (d') and response criteria index (beta). For this purpose, ten healthy female students (21.5+/-1.08 years) with a regular menstrual cycle participated in this study. A VDT-based visual inspection task was used for the experiment in 3x2 factorial designs. Two factors, menstrual phase, pre-menstruation (PMS), menstruation (M), and post-menstruation (PM) and background noise, with 80 dB(A) background noise and without noise, were analyzed as the main factors in this study. The results concluded that the sensitivity index (d') of SDT was affected in all the menstrual cycle conditions (p<0.01) and the presence of background noise (p<0.05) but no combinational effect of menstrual cycle and background noise was found in this study. On the other hand, no significant effect was observed in the subject's tendency in visual inspection, shown by beta along the menstrual cycle and the presence of background noise. According to the response criteria for each individual subject, the presence of noise affected the tendency of some subjects in detecting the object and making decision during the visual inspection task. PMID- 20009371 TI - Vaginal douching in Cambodian women: its prevalence and association with vaginal candidiasis. AB - BACKGROUND: We determined the prevalence of vaginal douching (cleansing of the vagina with liquid) in a sample of Cambodian women, and examined the associations of douching with genitourinary symptoms and infections, after controlling for potential confounding factors, including genitourinary symptoms and sociodemographic factors. METHODS: A total of 451 adolescent and adult females aged 15 to 49 years who attended 17 maternal and child health (MCH) clinics in 7 provinces of Cambodia in 2001 were consecutively enrolled as a part of the Sexually Transmitted Infection Sentinel Survey. Sociodemographic factors, genitourinary symptoms, and frequency of douching were assessed by face-to-face interviews using a structured questionnaire. Vaginal infections were examined by using standard diagnostic procedures specific to each pathogen. RESULTS: The proportion of participants who douched at least once a week was 76.7% (n = 346). Douching was significantly more prevalent in urban than in rural women (85.7%, n = 198 vs 67.3%, n = 148; P < 0.001). Frequency of douching was significantly associated with genitourinary symptoms, which were most prevalent in participants who douched from several times a week to once a day; genitourinary symptoms were less prevalent in those who douched more than once a day. Douching was significantly associated with vaginal candidiasis, but not with trichomoniasis or bacterial vaginosis, and this association persisted even after controlling for sociodemographic factors and genitourinary symptoms. CONCLUSIONS: Vaginal douching was very common among Cambodian women visiting MCH clinics. Further investigations are warranted to elucidate the reasons for douching. In addition, women should be informed that douching may endanger their reproductive health. PMID- 20009373 TI - Effect of chain wheel shape on crank torque, freely chosen pedal rate, and physiological responses during submaximal cycling. AB - The development of noncircular chain wheels for the enhancement of cycling performance has been in progress for a long time and continues apace. In this study we tested whether submaximal cycling using a non-circular (Biopace) versus a circular chain wheel resulted in lower peak crank torque at preset pedal rates as well as resulting in lower pedal rate and metabolic response at freely chosen pedal rate. Ten trained cyclists (mean+/-SD: 27+/-3 years of age, 182+/-4 cm tall, 77.5+/-7.0 kg of body mass, and peak oxygen uptake of 61.7+/-4.4 ml kg(-1) min(-1)) cycled with a Biopace and a circular chain wheel at 180 W at 65 and 90 rpm for recording of crank torque profiles, and at their freely chosen pedal rate for recording of pedal rate and metabolic response, including oxygen uptake and blood lactate concentration. Crank torque profiles were similar between the two chain wheels during cycling at preset pedal rates. During cycling at the freely chosen pedal rate (being 93+/-6 and 93+/-4 rpm for the Biopace and circular chain wheel, respectively), blood lactate concentration was significantly different between the two chain wheels, being on average 0.2 mmol l(-1) lower with the Biopace chain wheel. A musculoskeletal simulation model supported the idea that a contributing factor to the observed difference in blood lactate concentration may be slightly reduced muscle activity around the phase where peak crank torque occurs during cycling with the Biopace chain wheel. In that particular phase of the crank revolution, the observed slightly lower muscle activity may result from larger transfer of energy from the legs to the crank. PMID- 20009374 TI - Autonomic nervous activities assessed by heart rate variability in pre- and post adolescent Japanese. AB - There are many studies with respect to the age-related change of the characteristics of beat-to-beat heart rate variability (HRV), reflected by cardiac autonomic control, especially focusing on adulthood (i.e., aging related to the incidence of metabolic syndrome) in Japanese individuals. However, it is not still clear how basic control matures during childhood. This study was, therefore, designed to explore the HRV characteristics of pre- and post adolescent Japanese, in a cross-sectional manner. Resting HRV data was recorded in a relaxing supine position from 136 healthy individuals between 8 and 20 years (48 boys between 8 and 14 years; 88 girls between 8 and 20 years) who were instructed to breathe periodically (0.25 Hz). Frequency-domain analysis (i.e., the spectral analysis based on an autoregressive model) of short-term, stationary R-R intervals was performed to evaluate the low- (LF; below 0.15 Hz) and high- (HF; 0.15-0.40 Hz) frequency powers. The HF to total power represents the vagal control of heart rate (PNS indicator), and the ratio of LF to HF (LF/HF) is considered to relate to the sympathetic modulations (SNS indicator). Both PNS and SNS indices had substantially no effect from age and/or gender in the range between 8 and 20 years. In conclusion, the control of the cardiac autonomic nervous system in Japanese seems already to be compatible with that in adulthood before approximately 10 years. In other word, the cardiac autonomic modulation would presumably be maturated before the age of approximately 7-8 years, though further research is awaited. PMID- 20009375 TI - Individual variability in the core interthreshold zone as related to body physique, somatotype, and physical constitution. AB - For evaluating the effect of body physique, somatotype, and physical constitution on individual variability in the core interthreshold zone (CIZ), data from 22 healthy young Japanese male subjects were examined. The experiment was carried out in a climatic chamber in which air temperature was maintained at 20-24 degrees C. The subjects' body physique and the maximum work load were measured. Somatotype was predicted from the Heath-Carter Somatotype method. In addition, factors reflecting physical constitution, for example, susceptibility to heat and cold, and quality of sleep were obtained by questionnaire. The subjects wore a water-perfused suit which was perfused with water at a temperature of 25 degrees C and at a rate of 600 cc/min, and exercised on an ergometer at 50% of their maximum work rate for 10-15 min until their sweating rate increased. They then remained continuously seated without exercise until shivering increased. Rectal temperature (T(re)) and skin temperatures at four sites were monitored by thermistors, and sweating rate was measured at the forehead with a sweat rate monitor. Oxygen uptake was monitored with a gas analyzer. The results showed individual variability in the CIZ. According to the reciprocal cross-inhibition (RCI) theory, thermoafferent information from peripheral and core sensors is activated by T(re), mean skin temperature (T(sk)), and their changes. Since T(sk) was relatively unchanged, the data were selected to eliminate the influence of the core cooling rate on the sensor-to-effector pathway before RCI, and the relationship between the CIZ and the various factors was then analyzed. The results revealed that susceptibility to heat showed a good correlation with the CIZ, indicating that individual awareness of heat may change the CIZ due to thermoregulatory behavior. PMID- 20009376 TI - Daily physical activity in japanese preschool children evaluated by triaxial accelerometry: the relationship between period of engagement in moderate-to vigorous physical activity and daily step counts. AB - The purpose of the present study was to evaluate moderate-to-vigorous physical activity using triaxial accelerometry in Japanese preschool children. The relationship between daily step counts as a convenient measure of physical activity and minutes of engagement in moderate-to-vigorous physical activity was also examined. Physical activity was assessed using a triaxial accelerometer (ActivTracer, GMS) and daily steps using a uniaxial accelerometer for 6 consecutive days, including weekdays and weekend days, in 157 four- to six-year old Japanese children attending kindergarten or nursery school. Using triaxial and uniaxial accelerometers, nonlocomotive activities and step counts for young children can be evaluated, respectively. Average daily moderate-to-vigorous physical activity (physical activity ratio >or=3) and step counts were 102 (+/ 32) min/day and 13,037 (+/-2,846) steps/day, respectively. A strong and significant correlation was observed between minutes of moderate-to-vigorous physical activity and step counts (r=0.832, p<0.001). The daily step counts corresponding to 60 min, 100 min, and 120 min of moderate-to-vigorous physical activity were 9,934, 12,893, and 14,373 steps/day, respectively. The correlation coefficient between minutes of higher intensity activities (physical activity ratio >or=4) and step counts was slightly lower (r=0.604, p<0.001). The daily step count corresponding to 30 min of the higher intensity activities was 14,768 steps/day. These results suggest that approximately 13,000 steps/day are required for preschool children to engage in more than 100 min of moderate-to-vigorous physical activity. PMID- 20009377 TI - Dispersion-controlled slow light in photonic crystal waveguides. AB - Slow light with a markedly low group velocity is a promising solution for optical buffering and advanced time-domain optical signal processing. It is also anticipated to enhance linear and nonlinear effects and so miniaturize functional photonic devices because slow light compresses optical energy in space. Photonic crystal waveguide devices generate on-chip slow light at room temperature with a wide bandwidth and low dispersion suitable for short pulse transmission. This paper first explains the delay-bandwidth product, fractional delay, and tunability as crucial criteria for buffering capacity of slow light devices. Then the paper describes experimental observations of slow light pulse, exhibiting their record high values. It also demonstrates the nonlinear enhancement based on slow light pulse transmission. PMID- 20009378 TI - Extension of frequencies from maser to laser. How the laser evolved and was extended to terahertz during my research life: a personal review. AB - The present review describes the author's involvement and contributions to the development of the semiconductor laser and terahertz oscillators at his laboratory during the period between 1957 and now. The author cites records to show that the idea of a semiconductor laser was documented as a Japanese patent in April 1957 prior to those of G. Gould in 1957 and C.H. Townes in 1958. Terahertz oscillators of high Q values with the use of GaP were developed and applied to areas like investigations of molecular dynamics, cancer diagnosis, etc., thus extending the frontiers of science. PMID- 20009379 TI - Electrical conductivity of orthopyroxene: implications for the water content of the asthenosphere. AB - Electrical conductivity of minerals is sensitive to water content and hence can be used to infer the water content in the mantle. However, previous studies to infer the water content in the upper mantle were based on pure olivine model of the upper mantle. Influence of other minerals particularly that of orthopyroxene needs to be included to obtain a better estimate of water content in view of the high water solubility in this mineral. Here we report new results of electrical conductivity measurements on orthopyroxene, and apply these results to estimate the water content of the upper mantle of Earth. We found that the electrical conductivity of orthopyroxene is enhanced by the addition of water in a similar way as other minerals such as olivine and pyrope garnet. Using these new results, we calculate the electrical conductivity of pyrolite mantle as a function of water content and temperature incorporating the temperature and water fugacity dependent hydrogen partitioning. Reported values of asthenosphere conductivity of 4x10(-2)-10(-1) S/m corresponds to the water content of 0.01-0.04 wt%, a result in good agreement with the petrological model of the upper mantle. PMID- 20009380 TI - Heterogeneous electrical structure of Kozu-shima volcanic island, Japan. AB - Nearly twenty anomalous geoelectric field changes were observed before earthquakes at Kozu-shima Island, Japan, from 1997 to 2000. In order to help locating the current sources of the observed anomalous changes, a bipole-dipole resistivity survey was conducted. From the resistivity survey, including current injection into the ground, it was found that various features of the anomalous changes were systematically different from those of changes caused by artificial sources and induction of geomagnetic disturbances. Moreover, it is suspected that the currents of anomalous changes were generated not near the ground surface but deep under the ground. PMID- 20009381 TI - Development of newly designed VHF interferometer system for observing earthquake related atmospheric anomalies. AB - Temporal correlation between atmospheric anomalies and earthquakes has recently been verified statistically through measuring VHF FM radio waves transmitted beyond the line-of-sight. In order to locate the sources of such atmospheric anomalies, we developed a VHF interferometer system (bistatic-radar type) capable of finding the arrival direction of FM radio waves scattered possibly by earthquake-related atmospheric anomalies. In general, frequency modulation of FM radio waves produces ambiguity of arrival direction. However, our system, employing high-sampling rates of the order of kHz, can precisely measure the arrival direction of FM radio waves by stacking received signals. PMID- 20009382 TI - Recombinant alpha-actin for specific fluorescent labeling. AB - Until recently, actin was thought to act merely as a passive track for its motility partner, myosin, during actomyosin interactions. Yet a recent report having observed dynamical conformational changes in labeled skeletal muscle alpha actin suggests that actin has a more active role. Because the labeling technique was still immature, however, conclusions regarding the significance of the different conformations are difficult to make. Here, we describe the preparation of fully active alpha-actin obtained from a baculovirus expression system. We developed alpha-actin recombinants, of which subdomains 1 and 2 have specific sites for fluorescent probes. This specific labeling technique offers to significantly expand the information acquired from actin studies. PMID- 20009383 TI - Mechanism of cadmium induced crystal defects in developing rat tooth enamel. AB - It is well known that exposure to environmental cadmium causes itai-itai (ouch ouch) disease. However, the exact mechanism underlying this bone disease remains unresolved. By focusing on the calcification mechanism, we examined developing tooth enamel in rats exposed to cadmium to test the hypothesis that cadmium exposure may cause defects in crystal formation. Electron microscopy revealed the presence of perforated crystals in developing tooth enamel, indicating that the process of crystal nucleation may have been interrupted by cadmium exposure. Furthermore, biochemical analyses revealed that the catalytic activity of carbonic anhydrase in the immature enamel matrix declined remarkably despite the fact that quantitative reduction of this enzyme was insignificant, suggesting that the decline of catalytic activity may have resulted from the replacement of zinc with cadmium ions. Therefore, we concluded that the poor catalytic activity of cadmium-binding carbonic anhydrase might hinder the nucleation process, leading to an impairment in mineralization that causes itai-itai disease. PMID- 20009384 TI - [Case report of an attitude survey contributing to the improvement of psychological problems in an organization]. AB - When using attitude surveys to improve the psychological problems in an organization, its main objective is to provide material for organizational decision-making by exploring the necessity of countermeasures; and if countermeasures are necessary, the objective is to illustrate the factors behind the issues quantitatively. In this case study, we attempted to clarify the necessity of the organization to address the psychological problems its members carried; In order to find effective countermeasures, an attitude survey was performed. The questionnaire consisted of general survey items and items uniquely prepared for the organization. Based on the results of the questionnaire, countermeasures were proposed. After the proposed countermeasures were implemented, a follow-up investigation was conducted to evaluate the effect. Acting upon a request from a labor union, we conducted interviews at a regional plant of a major manufacturing company where many employees carry out strenuous tasks and sick leave due to mental illness is a frequent occurrence. Then, a questionnaire-based survey that reflected the results of the interviews was performed. Analyses of variance of the general survey items showed that the mental health and morale of the union members were significantly poor. Also, a comparison of structure of consciousness using a multi-group structural equation analysis and a comparison with the survey items uniquely prepared for the organization indicated that a dehumanizing workload and a negative environment created by the high turnover rate and confusion in the chain of command had alienated the employees. These results were reported to the management by the union and they jointly implemented some countermeasures. A year later, a second survey that included items that reflected the countermeasures taken by the organization was conducted. Analyses of variance showed a significant improvement in the results, and multi-regression analysis using the aggregated data (baseline and post-intervention) showed that some of the countermeasures taken by the union and the management were probably effective. The results suggest that, when the party conducting the investigation and the party implementing the countermeasures are different, it would be effective to devise an investigation and a statistical analysis that can unravel the actual status of the organization based on the empirical findings, since it would provide useful materials for organizational decision-making and validation of the effects of the implemented countermeasures. PMID- 20009385 TI - [Results of a questionnaire survey about the activities of the Bureau of Occupational Physicians in the Japan Society for Occupational Health]. PMID- 20009386 TI - [Influence of physical workload patterns and breaks on heart rate recovery]. AB - It is necessary to try to achieve quick recovery from work strain by setting adequate breaks and shortening continuous working hours to prevent the accumulation of fatigue. However, there has been no research investigating the influence of the timing and lengths of breaks on individual aerobic capacities in recovery from work strain. In this study, we set three load patterns based on the length and timing of breaks: "no breaks", "one break" and "regular small breaks". We examined the differences of the heart rate variation in the recovery time after working considering the individual aerobic capacities (VO(2)max) of ten male subjects (mean age 22.3 +/- 1.7 yr) in the case of 50 W or 100 W workloads on a bicycle ergometer. When individual aerobic capacity was not considered, the "regular small breaks" condition led to the quickest recovery to the level of the resting heart rate at 50 W workload. Not all conditions showed heart rate recovery within 30 min at 100 W workload. On the other hand, when individual aerobic capacity was considered, the "regular small breaks" condition showed the quickest recovery to the level of the resting heart rate at 50 W workload in the low aerobic capacity group (VO(2)max mean 42.2 +/- 3.7 ml/kg/min). However, in the high aerobic capacity group (VO(2)max mean 54.5 +/- 4.1 ml/kg/min), the "regular small breaks" condition resulted in the quickest recovery of the level to the resting heart rate at 100W workload. Two-way repeated measures ANOVA was performed for the recovery time with respect to the rate of increase from resting heart rate to examine the influence on heart rate recovery of physical activity loads, workload patterns and individual fitness. Physical activity loads were strongly related to the increase from resting heart rate in recovery time, and workload patterns showed that the regular small breaks condition was related to the heart rate recovery in the high fitness subjects in the case of the exercise intensity of 100 W. The effect of work strain decreased by workload patterns was defined by the relationship between physical activity load and individual fitness. In future occupational health activities, it will be important to manage work in order to aim for quick recovery from work strain by using ergonomics. The evaluation of the level of the physical activity load and distribution of proper break times during working in addition to individual aerobic capacities are important in considerations of quick recovery from work strain. PMID- 20009387 TI - Recruitment of immune cells across atrial endocardium in human atrial fibrillation. AB - BACKGROUND: Although clinical studies have suggested a link between inflammation markers and atrial fibrillation (AF), it is still unclear whether local immunologic responses actually exist in human atria during AF. METHODS AND RESULTS: To address this point, human left appendages were obtained from 16 patients who underwent cardiac surgery (5 with sinus rhythm (SR) and 11 with AF) and subjected to immunohistochemical analysis. In all the AF specimens, adhesion and migration of CD45-reactive cells were consistently observed predominantly in the atrial endo- and subendomyocardium and more prominently than in SR. Most of them were immunologically active CD68-positive macrophages, whereas CD3-positive T cells infiltrated to a lesser extent. Scavenger-receptor A staining revealed maturation of macrophages not in the endocardium but in the midmyocardium, a gradient from endo- to midmyocardium. In the endocardium, along with adhesion molecules (intracellular adhesion molecule-1 and vascular cell adhesion molecule 1), a chemotactic protein-1, which facilitates the recruitment, was more abundantly expressed in AF than in SR. Cytokines including transforming growth factor-beta and interleukin-6 were frequently expressed by these macrophages. CONCLUSIONS: These observations collectively imply active adhesion and recruitment of macrophages across the endocardium in human fibrillating atria, thereby supporting the concept of local immunologic inflammatory responses around the atrial endocardium of AF. PMID- 20009388 TI - Evaluation of subclinical right ventricular dysfunction in obstructive sleep apnea patients using velocity vector imaging. AB - BACKGROUND: The aims of this study were to evaluate subclinical regional right ventricular (RV) dysfunction in newly diagnosed obstructive sleep apnea (OSA) patients without systemic and pulmonary arterial (PA) hypertension, and to correlate OSA severity to RV dysfunction, using both velocity vector imaging (VVI)-derived strain imaging and tissue Doppler imaging (TDI). METHODS AND RESULTS: The OSA group consisted of 27 patients and the control group consisted of 26 healthy participants. All participants underwent 24-h ambulatory blood pressure monitoring. Peak systolic myocardial velocities, strain, and strain rate (SR) were determined at the basal and mid segments of the RV free wall by VVI. Additionally, RV myocardial velocities were assessed by pulsed-wave TDI. Patients with OSA had significantly impaired VVI-derived peak systolic myocardial velocities, strain, and SR (P<0.0001 for all). RV isovolumic acceleration (IVA) was the only TDI-derived parameter that was significantly impaired (P<0.0001). RV IVA (r=-0.512, P<0.0001), RV mid free wall strain (r=0.568, P<0.0001) and SR (r=0.519, P<0.0001) revealed the best correlations with apnea hypopnea index (AHI). CONCLUSIONS: Subclinical RV dysfunction is present in OSA patients despite normal systemic and PA pressures. Tissue Doppler-derived RV IVA and VVI-derived RV deformation can accurately recognize and quantify RV function abnormalities in this subgroup of patients. PMID- 20009389 TI - Age-induced reduction in mitochondrial manganese superoxide dismutase activity and tolerance of macrophages against apoptosis induced by oxidized low density lipoprotein. AB - BACKGROUND: Oxidative stress is thought to play an important role in age-induced atherogenesis. Manganese superoxide dismutase (MnSOD) is an antioxidant enzyme that is localized in mitochondria and protects macrophages against apoptosis induced by oxidized low density lipoprotein (oxLDL). We previously reported that genetic polymorphism of MnSOD modifies mitochondrial MnSOD (mtMnSOD) activity and increases the risk of coronary artery disease. In this study, we investigated the association of mtMnSOD activity with aging. METHODS AND RESULTS: Blood samples were taken from 69 healthy participants aged 20-52. The MnSOD genotype was analyzed using real-time polymerase chain reaction. Leukocyte mtMnSOD activity was measured by inhibition of WST-1. Macrophages were treated with oxLDL and the apoptotic cells were counted. mtMnSOD activity was inversely correlated with the age of the participant regardless of the MnSOD genotype. The percentage of apoptotic macrophages after incubation with oxLDL correlated with age. Thus, the percentage of apoptotic macrophages after incubation with oxLDL was inversely related to mtMnSOD activity. Lecithinized SOD, which can easily transfer into cells, improved the tolerance of macrophages against oxLDL. CONCLUSIONS: mtMnSOD activity decreases with age, thereby reducing the tolerance of macrophages against oxLDL-induced apoptosis. Our data may provide an important clue to clarify the mechanisms of age-induced atherosclerosis. PMID- 20009390 TI - Autoimmune thyroid disease in children and adolescents with idiopathic pulmonary arterial hypertension. AB - BACKGROUND: Previous studies have reported a high prevalence of autoimmune thyroid disease (AITD) in adult patients with pulmonary arterial hypertension (PAH). The aim of this retrospective study was to determine the prevalence of AITD in children and adolescents with idiopathic PAH (IPAH). METHODS AND RESULTS: The study group included 16 patients who had been diagnosed as having idiopathic PAH when they were younger than 15 years old; all were younger than 20 years of age. Thyroid function and antithyroid antibody levels were examined regularly at 6-12-month intervals and when there were clinical signs of thyroid dysfunction. In total, 7 patients (44%) had AITD; 2 patients developed Graves' disease, 2 developed silent thyroiditis, and 3 had antithyroid antibodies with euthyroidism. The duration after PAH onset and the prostacyclin (PGI(2)) treatment period were significantly longer in patients with AITD (7.6+/-2.1 and 7.4+/-2.3 years, respectively) than in patients without AITD (5.0+/-1.1 and 4.8+/-1.2 years, respectively; P<0.01 and P<0.05). CONCLUSIONS: The prevalence of AITD is high in children and adolescents with IPAH, so evaluation of thyroid function is important to prevent deterioration of right heart failure. PMID- 20009391 TI - Tricuspid annuloplasty using the MC3 ring for functional tricuspid regurgitation. AB - BACKGROUND: A recently introduced tricuspid annuloplasty ring, the MC(3) ring, has a 3-dimensional form that is designed to remodel the tricuspid valve annulus. The aim of this study was to investigate its clinical performance. METHODS AND RESULTS: From December 2004 to April 2008, 103 patients underwent tricuspid annuloplasty using the MC(3) ring (mean age, 52+/-13 years; 63.6% women). The average preoperative tricuspid regurgitation (TR) grade was 2.5+/-0.8, and the mean preoperative systolic pulmonary artery pressure was 48.4+/-15.0 (24-88) mmHg; the mean follow-up was 26.7+/-11.2 (0-52) months. One patient died after surgery (1.0%), because of cor pulmonale. No MC(3) ring-related complications, such as, atrioventricular block, ring dehiscence or thromboembolism, were encountered. Predischarge echocardiography showed a significant decrease in the TR grade (2.5+/-0.8 to 0.8+/-0.8, respectively; P<0.001). After a median 15 months, the mean TR grade was stable (0.9+/-0.8). The mean systolic pulmonary artery pressure was also lower than its preoperative value (33.9+/-7.9 vs 48.4+/ 15.0 mmHg, respectively; P<0.001). CONCLUSIONS: The MC(3) ring provides good mid term clinical and echocardiographic results for TR. However, long-term follow-up is mandatory to confirm the stability of this procedure. PMID- 20009392 TI - The influence of Helicobacter pylori eradication on leptin, soluble CD40 ligand, oxidative stress and body composition in patients with peptic ulcer disease. AB - OBJECTIVE: To determine the effect of Helicobacter pylori (H. pylori) eradication on blood levels of soluble CD40 ligand, leptin, oxidative stress and body composition in patients with dyspepsia infected with H. pylori. METHODS: The infection of H. pylori was based on the presence of both (14)C urea breath test (UBT) and histology. Patients were given triple eradication therapy for 14 days and at 3 months after the treatment, (14)C UBT was reinstituted. Fasting glucose, leptin, body composition, soluble CD40 ligand, total oxidant status (TOS) were studied before and at 3 months after the treatment. RESULTS: In 33 subjects, H. pylori infection was successfully eradicated. sCD40L, and TOS levels were significantly decreased after H. pylori eradication. The percentage of body fat and body fat mass significantly decreased whereas the fat free mass (FFM) increased after eradication. However, eradication of the organism yielded no differences in leptin levels. CONCLUSION: These findings suggest that H. pylori eradication reduces the sCD40L and oxidative stress, fat mass with a significant increase in fat free mass. Thus, eradication of H. pylori infection not only improves ulcer healing, but may also reduce the presumed atherosclerosis risk. PMID- 20009393 TI - Active tuberculosis in patients undergoing hemodialysis for end-stage renal disease: a 9-year retrospective analysis in a single center. AB - OBJECTIVE: Tuberculosis (TB) in patients undergoing hemodialysis (HD) for end stage renal disease (ESRD) is commonly thought to be associated with a very poor prognosis. Moreover, it is difficult to diagnose. This report was designed to describe this condition and to determine the mortality rate and risk factors associated with mortality. In addition, the study evaluated the usefulness of QuantiFERON TB-2G((R)) (QFT-2G). METHODS: Retrospective study PATIENTS: Patients with confirmed TB admitted between January 2001 and May 2009 were retrospectively identified and enrolled. The clinical, radiological, and bacteriological data at the time of admission were recorded. A multivariate analysis was performed to identify the predictive factors for mortality. RESULTS: A total 19 TB patients (6 females; median age, 73 years) were included. TB occurred in most cases within 1.3 years from the initiation of dialysis. Most patients presented with fever (84.2%) and extrapulmonary TB (57.9%). The mortality rate within 24 weeks of the initiation of TB treatment was 36.8%. The factors associated with mortality were: a short duration of dialysis (HR 8.86, 95% CI 1.03-75.7, p=0.04), and underweight (HR 10.88, 95% CI 1.28-92.6, p=0.02). The sensitivity of QFT-2G, acid-fast smear, and polymerase chain reaction was 50, 80, and 88.2% respectively. CONCLUSION: These data indicate a high incidence of TB in the early stages of HD and a high mortality rate among these patients. The clinical utility of QFT-2G was found to be limited. Hypoalbuminemia might therefore be related to either indeterminate or negative results of QFT-2G. PMID- 20009394 TI - Moxifloxacin-based triple therapy versus clarithromycin-based triple therapy for first-line treatment of Helicobacter pylori infection: a meta-analysis of randomized controlled trials. AB - BACKGROUND: Moxifloxacin-based triple therapy has been suggested as an alternative first line therapy to clarithromycin-based triple therapy for Helicobacter pylori infection. AIMS: To systematically review the efficacy and tolerance of moxifloxacin-based triple therapy, and to conduct a meta-analysis of studies comparing this regimen with clarithromycin-based triple therapy. METHODS: A search of The Cochrane Library, PUBMED, EMBASE, EBM Review databases, Science Citation Index Expanded, and CMB (Chinese Biomedical Literature Database) was performed. Randomized controlled trials comparing moxifloxacin-based triple therapy to gold standard triple therapy in the first-line treatment of Helicobacter pylori infection were selected for meta-analysis. Relative risk was used as a measure of the effect of the two above-mentioned regimens with a fixed effects model using the methods of DerSimonian and Laird. RESULTS: Four randomized controlled trials totaling 772 patients were included. The meta analysis showed that the mean eradication rate was 84.1 (318/378) in the moxifloxacin-based triple therapy group and 73.6 (290/394) in the clarithromycin based triple therapy group; there was statistical significance between the two groups (RR, 1.13; 95% CI, 1.01, 1.27; P=0.04). There were no statistically significant difference in the overall side effects (RR, 0.61; 95% CI, 0.25, 1.48; P<0.28). CONCLUSIONS: Moxifloxacin-based triple therapy is more effective and does not increase the incidence of overall side effects compared to clarithromycin-based triple therapy in the treatment of H. pylori infection. PMID- 20009395 TI - The sky blue method as a screening test to detect misplacement of percutaneous endoscopic gastrostomy tube at exchange. AB - BACKGROUND: During tube exchange for percutaneous endoscopic gastrostomy (PEG), a misplaced tube can cause peritonitis and death. Thus, endoscopic or radiologic observation is required at tube exchange to make sure the tube is placed correctly. However, these procedures cost extensive time and money to perform in all patients at the time of tube exchange. Therefore, we developed the "sky blue method" as a screening test to detect misplacement of the PEG tube during tube exchange. METHODS: First, sky blue solution consisting of indigocarmine diluted with saline was injected into the gastric space via the old PEG tube just before the tube exchange. Next, the tube was exchanged using a standard method. Then, we checked whether the sky blue solution could be collected through the new tube or not. Finally, we confirmed correct placement of the tube by endoscopic or radiologic observation for all patients. RESULTS: A total of 961 patients were enrolled. Each tube exchange took 1 to 3 minutes, and there were no adverse effects. Four patients experienced a misplaced tube, all of which were detectable with the sky blue method. Diagnostic parameters of the sky blue method were as follows: sensitivity, 94% (95%CI: 92-95%); specificity, 100% (95%CI: 40-100%); positive predictive value, 100% (95%CI: 100-100%); negative predictive value, 6% (95%CI: 2-16%). CONCLUSION: These results suggest that the number of endoscopic or radiologic observations to confirm correct replacement of the PEG tube may be reduced to one fifteenth using the sky blue method. PMID- 20009396 TI - Perivascular epithelioid cell tumor (PEComa) of the liver diagnosed by contrast enhanced ultrasonography. AB - Perivascular epithelioid cell (PEC) is a unique cell which expresses both myogenic and melanocytic markers, and forms PEComa. A 36-year-old woman presented with a 35 mm-diameter liver tumor. MRI showed poor fat component in the tumor. Contrast-enhanced ultrasonography using the newly developed enhancing reagent, Sonazoid, clearly demonstrated early-phase enhancement of the tumor and rapid drainage of the reagent to veins, suggesting a PEComa. Lateral segmentectomy of the liver was performed. Histologically, epithelioid tumor cells around the vessels were immunostained with both HMB-45 and alpha-smooth muscle actin, confirming the diagnosis of PEComa. No recurrence has been found for 18 months following the operation. PMID- 20009397 TI - Autoimmune pancreatitis without a response to steroid therapy: a case which met criteria after withdrawal of steroid. AB - Autoimmune pancreatitis (AIP) sometimes forms a pancreatic mass lesion, which is often difficult to distinguish from pancreatobiliary malignancy, however it generally responds to steroid therapy. A 70-year-old man was referred to our institute with the suspected diagnosis of pancreatic cancer due to a mass lesion detected at the pancreatic head. Various images demonstrated an ill-defined mass at the enlarged pancreatic head with focal narrowing of the main pancreatic duct. Serum antinuclear antibody (ANA) was negative (x40 dilution) on the onset. Forceps biopsy from the narrowed pancreaticobiliary duct and fine-needle aspiration biopsy under endoscopic ultrasonography (EUS-FNAB) ruled out pancreatobiliary malignancy. Steroid therapy was started at 40 mg per day but was not effective according to subsequent image analyses. Repeated EUS-FNA from the pancreatic mass was performed but was again negative for carcinoma. Seven months later, under steroid-off condition, still no response was recognized in the clinical image but the titer of serum ANA was increased to be positive (x80), satisfying the criteria of AIP in Japan (2006). Although very rare, this is a case meeting Japanese criteria of AIP after withdrawal of steroid without response to steroid in the clinical images, suggesting the necessity of careful follow-up. PMID- 20009398 TI - Diffuse liver infiltration by melanoma of unknown primary origin: one case report and literature review. AB - Diffuse liver infiltration by melanoma of unknown primary origin is rare. We encountered a unique case of diffuse liver infiltration by melanoma of unknown primary origin in our hospital. A 62-year-old woman was referred to our hospital for anorexia of 6 months duration and abdominal distension for 1 month. Ultrasonography (US), computerized tomography (CT) and magnetic resonance imaging (MRI) revealed an obvious enlarged liver without detectable nodules. She was diagnosed as liver metastasis by melanoma of unknown primary origin via percutaneous liver biopsy. The report demonstrates the difficulty of making a noninvasive diagnosis of diffuse hepatic infiltration on metastatic melanoma. PMID- 20009399 TI - Cholangiocarcinoma producing parathyroid hormone-related peptide treated with chemoradiation using gemcitabine and S-1. AB - Although chemotherapy for cholangiocellular carcinoma (CCC) is administered to those patients who are inoperable, the results are largely disappointing, especially for CCC producing parathyroid hormone-related peptide (PTHrP). We encountered a 43-year-old man with hypercalcemia due to PTHrP secretion of CCC. As the tumor was inoperable, we treated him with gemcitabine (GEM), S-1 and radiation. The tumor size was reduced 55% and the serum PTHrP level decreased markedly after the chemoradiotherapy. Although the patient died after 14 months, this case clearly suggests that the combined chemoradiotherapy of GEM, S-1 and radiation, may be useful for the treatment of CCC producing PTHrP. PMID- 20009400 TI - Right atrial thrombosis after upgrading to a biventricular pacing/defibrillation system. AB - A 56-year-old man under right ventricular pacing for atrial fibrillation and bradycardia had congestive heart failure. He received a cardiac resynchronization pacemaker with a defibrillator. Four months later, follow-up transthoracic echocardiography showed a right atrial mass although he had no symptom. Transesophageal echocardiography showed a large immobile round-shaped mass on the defibrillation lead, which was attached to the free wall of the right atrium. One month after the initiation of anticoagulant therapy, the mass disappeared, suggesting that it was thrombotic. During the 5 month follow-up, he remained in good condition without the recurrence of right atrial thrombosis. PMID- 20009401 TI - Overtly manifested diabetes mellitus after resection of insulinoma. AB - Insulinoma is the most common cause of endogenous hyperinsulinemic hypoglycemia in adults. However, the coincidence of insulinoma and diabetes is extremely uncommon. We describe a rare, but very interesting case of diabetes mellitus which was masked by insulinoma and was overtly manifest after the removal of the insulinoma. PMID- 20009402 TI - Adrenal failure caused by a retroperitoneal malignant mesothelioma. AB - A 62-year-old man presented with clinical signs of acute abdominal pain and adrenal insufficiency. Computerized tomographic scans revealed bilateral adrenal tumors and the left adrenal tumor was surgically resected. The patient was found to have retroperitoneal mesothelioma presenting as bilateral metastatic adrenal tumors, which were well controlled by systemic chemotherapy with CDDP and 5-FU. This case appears to be the first reported case in the literature of adrenal failure due to bilateral adrenal infiltration caused by retroperitoneal malignant mesothelioma. PMID- 20009403 TI - A spontaneous spinal epidural hematoma in a hemodialysis patient: a rare entity. AB - Hemodialysis patients are at an increased risk of bleeding due to the platelet dysfunction caused by uremia and the use of anticoagulants during dialysis. Spontaneous spinal hematoma is a rare disorder as a complication in hemodialysis patients. Also it includes the hematoma secondary to coagulopathy, vascular malformation and hemorrhagic tumors. Here, we report the case of 77-year-old woman who presented with spinal cord compression due to spontaneous spinal epidural hematoma associated with hemodialysis. When an end-stage renal disease patient suffers from back pain and neurological deficits, the clinician should be alerted for the spontaneous spinal epidural hematoma as well as cerebrovascular events. PMID- 20009404 TI - Hemorrhagic shock and obstructive uropathy due to a large rectus sheath hematoma in a patient on anticoagulant therapy. AB - A 54-year-old woman was transferred to our hospital with disseminated intravascular coagulation, and was treated with heparin. On hospitalization day 13, she developed lower abdominal pain and mass followed by circulatory shock. She became oliguric and laboratory tests showed serum creatinine of 3.5 mg/dL and hemoglobin of 7.4 g/dL. Computed tomography showed hematoma in the left rectus sheath, compressing the urinary bladder exteriorly, which resulted in worsening of bilateral hydronephrosis. Conservative treatment resulted in resolution of the rectus sheath hematoma and improvement of renal function. Rectus sheath hematoma can be treated conservatively without surgical intervention even in complicated cases. PMID- 20009405 TI - Bilateral phrenic nerve paralysis manifested by orthopnea for 6 months in a patient with neuralgic amyotrophy. AB - Bilateral phrenic nerve paralysis (BPP) is a relatively rare disease manifested by slight dyspnea at rest and on exertion in the sitting and standing positions and by dyspnea in the supine position. A 67-year-old man, who was a painter, presented with severe pain in both shoulder regions that had evolved into orthopnea and forced him to sleep in a sitting position at night. Dyspnea and paradoxical respiratory movement in the supine position raised suspicions of BPP. The most striking feature in this case was that the rapid onset of pain in both shoulder regions was followed by BPP. The BPP was considered to be secondary to neuralgic amyotrophy (NA). PMID- 20009406 TI - Germ cell tumor concomitant with acquired immune deficiency syndrome. AB - Patients with acquired immune deficiency syndrome (AIDS) are susceptible to secondary malignant tumors. Among those malignancies, the increased incidence of germ cell tumor (GCT) in patients with AIDS has recently been documented in Western countries, while that is still rare in Japan. Here, we report a man patient with advanced GCT (seminoma) complicated with AIDS who was continuously treated with highly active antiretroviral therapy (HAART). A partial response was obtained after resection of the primary left testis and three courses of chemotherapy. During the clinical course, he contracted unexpected gastric bleeding that made it impossible to take HAART agents and prophylactic agents for opportunistic infection. Thereafter, he suffered from a severe pulmonary infection and consequently died of severe respiratory failure. The lymphopenia related to both chemotherapy and AIDS synergistically rendered this patient immunoincompetent and thus he suffered from this fatal pulmonary infection. The recent progress in AIDS treatment has been reported to prolong the survival of tumor-bearing AIDS patients, especially GCT-bearing AIDS patients. Because of the current increase in the number of AIDS patients in Japan, it is important to report the present case which indicated that careful chemotherapy against GCT with strict management of the immunoincompetence can provide a good prognosis for GCT-bearing AIDS patients. PMID- 20009407 TI - The middle meningial artery during a migraine attack: 3T magnetic resonance angiography study. AB - We performed 3T magnetic resonance angiography (MRA) during a spontaneous migraine attack. The patient was a 42-year-old woman migraineur diagnosed by the IHS criteria. The change of the middle meningial artery (MMA) was measured on the axial brain images using MATLAB for three phases (attack-free period, during an attack, a period after medication). There were no dramatic changes of vasodilation in the MMA during the attack (2.0 mm), attack-free period (diameter 1.9 mm), or period after medication (1.7 mm), resembling extrapolations of observations in experimental animal models. This finding suggests that the dramatic vasomotion might not be associated with migraine pathophysiology. PMID- 20009408 TI - Severe hyperthermia caused by four-vessel occlusion of main cerebral arteries. AB - We present the case of a comatose patient with acute large infarction of posterior cerebral and cerebellar areas and severe hyperthermia (max. 40.4 degrees C). Angiography demonstrated four-vessel occlusion of the main cerebral arteries, suggesting the possibility that both internal carotid and left vertebral arteries were already occluded and he became unconscious following additional occlusion of the right vertebral artery. Autopsy findings revealed bilateral ischemic damage of the hypothalamus in addition to the above infarct areas. Sudden ischemic involvement of both hypothalamic regions may have caused the extremely high fever in this case. PMID- 20009409 TI - Short-lasting unilateral neuralgiform headache attacks with cranial autonomic symptoms (SUNA). AB - Short-lasting unilateral neuralgiform headache attacks with cranial autonomic symptoms (SUNA) is a rare primary headache syndrome. The diagnostic criteria include attacks of unilateral orbital, supraorbital or temporal stabbing pain accompanied by one of the following: conjunctival injection and/or tearing, nasal congestion and/or rhinorrhea, and eyelid edema. The duration of pain is 2 seconds to 10 minutes, and the frequency of attacks is described as once a day or more. The etiology and pathology of SUNA has yet to be documented. We report an 18-year old man with SUNA. Lomerizine hydrochloride which is used as a preventive medicine for migraine, improved his headaches. PMID- 20009410 TI - CMV infection presenting as a cavitary lung lesion in a patient with systemic lupus erythematosus receiving immunosuppressive therapy. AB - We report a case of cytomegalovirus (CMV) pneumonitis that presented as a cavitary lung lesion in a patient with systemic lupus erythematosus receiving immunosuppressive treatment. The lesion was confirmed by positive polymerase chain reaction (PCR) for CMV in bronchoalveolar lavage fluid (BALF) and CMV antigenemia. PCR for CMV in BALF was demonstrated to be useful for the diagnosis of CMV pneumonitis on the basis of high sensitivity and specificity. After initiating ganciclovir, the lesion gradually regressed. A cavitary lung lesion associated with CMV is extremely rare. This presentation suggests that the differential diagnosis of cavitary lung lesion in immunocompromised individuals should include CMV. PMID- 20009411 TI - Multiple lobulated cystic lesions in suprasellar cistern. PMID- 20009412 TI - Dissecting intramural hematoma of the esophagus in a Kendo player taking low-dose aspirin. PMID- 20009413 TI - Intravascular lymphoma: usefulness of liver biopsy in the clinical setting. PMID- 20009414 TI - Primary bone lymphoma of the scapula. PMID- 20009415 TI - Pulmonary artery pseudoaneurysm secondary to lung abscess. PMID- 20009416 TI - The lectin-like oxidized low-density lipoprotein receptor-1 and its soluble form: cardiovascular implications. AB - The lectin-like oxidized low density lipoprotein receptor-1 (LOX-1) is a multiligand receptor, whose repertoire of ligands includes oxidized low-density lipoprotein, advanced glycation endproducts, platelets, neutrophils, apoptotic/aged cells and bacteria. Sustained expression of LOX-1 by critical target cells, including endothelial cells, smooth muscle cells and macrophages in proximity to these ligands, sets the stage for chronic cellular activation and tissue damage suggesting the interaction of cellular LOX-1 with its ligands to contribute to the formation and development of atherosclerotic plaques. Studies with transgenic and knockout mouse models have elucidated in part the role of LOX 1 in the pathogenesis of atherosclerosis and cardiac remodeling. Recently, a circulating soluble form of LOX-1 (sLOX-1), corresponding solely to its extracellular domain, has been identified in human serum. Circulating levels of sLOX-1 are increased in inflammatory and atherosclerotic conditions and are associated with acute coronary syndrome, with the severity of coronary artery disease, and with serum biomarkers for oxidative stress and inflammation, suggesting that they could be a useful marker for vascular injury. However, many interesting questions have not yet been answered and in this review, we provide an updated overview of the literature on this receptor and on likely future directions. PMID- 20009417 TI - Serum cholinesterase levels of Thai chilli-farm workers exposed to chemical pesticides: prevalence estimates and associated factors. AB - OBJECTIVE: To estimate the prevalence of, and factors associated with, abnormal serum cholinesterase (SChE) levels among chilli-farm workers in Chatturat District, Chaiyaphum Province. METHODS: A total of 350 chilli-farm workers aged 18-60 yr were randomly sampled and interviewed. A reactive-paper finger-blood test was used to assess SChE levels. RESULTS: The prevalence of abnormal SChE levels was 32.0%. The most common pesticide-related symptoms were dizziness (38.0%), headache (30.9%), nausea/vomiting (26.9%), and fever (26.9%). Multiple logistic regression analysis revealed 7 factors were independently associated with abnormal SChE level: male gender, single/separated/divorced, being a permanent worker, spraying pesticide more than 3 times per month, having moderate or poor pesticide-use behaviors, and low perceived susceptibility and severity of pesticide use. CONCLUSIONS: The prevalence of abnormal SChE levels among chilli farm workers was quite high. It would be beneficial to decrease pesticide use and encourage alternative measures. Effective preventive interventions to increase correct perceptions of pesticide use, the use of personal protective measures and continuing monitoring for blood cholinesterase, especially for male permanent farm workers, are recommended. PMID- 20009418 TI - Matrix metalloproteinase-3 and vitamin d receptor genetic polymorphisms, and their interactions with occupational exposure in lumbar disc degeneration. AB - OBJECTIVES: To investigate the occupational and genetic risk factors inducing lumbar disc degeneration in a Chinese population, and to explore their synergistic interactions. METHODS: A case-control study involving 178 low back pain patients with lumbar disc degeneration and 284 controls was carried out. Five types of work-related factors were investigated using questionnaires. Polymerase chain reaction and restriction fragments length polymorphism was used to detect the polymorphisms of MMP-3 (matrix metalloproteinase-3)(rs731236), VDR Taq (vitamin D receptor-Taq) and VDR-Apa (vitamin D receptor-Apa)(rs35068180). Rothman's synergy index was used to measure the synergistic interactions between gene polymorphisms and occupational risk factors. RESULTS: Family history of lumbar disc diseases, back injury history, whole-body vibration, bending/twisting, heavy physical workload, age, mutation alleles 5A of MMP-3 and A of VDR-Apa were significantly associated with lumbar disc degeneration (OR=12.70, 11.79, 8.96, 5.46, 1.05, 1.96 and 1.70, respectively, p<0.05). Synergistic interactions existed between the mutation allele 5A of MMP-3 and whole-body vibration exposure, the mutation allele 5A of MMP-3 and bending/twisting, and the mutation allele A of VDR-Apa and bending/twisting (SI=13.27, 2.91, 2.35, respectively, p<0.05). CONCLUSIONS: The results of this study suggest that gene-occupation interaction might play a certain role in exaggerating lumbar disc degeneration. There is a possibility that subjects who carry mutation alleles 5A of MMP-3 and/or A of VDR-Apa are more vulnerable to lumbar disc degeneration when they are exposed to whole-body vibration and/or bending/twisting under ergonomic loads. PMID- 20009419 TI - Important behavioral traits for predicting guide dog qualification. AB - Guide dogs for the blind help blind people physically and mentally in their daily lives. Their qualifications are based on health, working performance and temperament; approximately 70% of dogs that fail to qualify are disqualified for behavioral reasons. In order to achieve early prediction of qualification, it would be essential as the first step to identify important temperament traits for guide dogs. Therefore, we administered a questionnaire consisting of 22 temperament items to experienced trainers to assess candidate dogs at the Japan Guide Dog Association after three months of training, which was at least three months prior to the final success (qualified as a guide dog) or failure (disqualified for behavioral reasons) judgment. Factor analyses of question items stably extracted three factors with high internal consistency, Distraction, Sensitivity and Docility. When we compared factor points between successful dogs and failed dogs, the successful dogs showed significantly and consistently lower Distraction points and higher Docility points. Additionally, Distraction points could predict qualification with 80.6% accuracy and detect 28.2% of the failed dogs that had higher Distraction points than any of the successful dogs. Of the nine question items not included in the three factors, two items (;Aggression' and ;Animal interest') were consistently associated with qualification. These results suggest that Distraction is stably assessable and has the strongest impact on success or failure judgment; therefore, it will be the first target to establish a behavioral test that may lead to early prediction of guide dog qualification. PMID- 20009420 TI - Hypertension after ingestion of baked garlic (Allium sativum) in a dog. AB - A 6-year-old, intact male Schnauzer was referred 2-days after accidental ingestion of baked garlic. Regenerative anemia (Hematocrit 22%) and the elevated methemoglobin (8.7%) concentration were detected upon hematological examination. Eccentrocytes, Heinz bodies and ruptured red blood cells were also noted on blood smear films, which were the results from the oxidative injury of the Allium species. The dog was hypertension (systolic mean 182 mmHg) concurrent with other clinical signs, such as vomiting and dark brown urination. Treatment with continuous oxygen, antioxidant drugs and antihypertensive therapy resulted in good progress. The dog was discharged 4 days after hospitalization. There were no remarkable findings in the follow up hematologic examination 24 days after discharge, but the dog still had a high blood pressure and continued on antihypertensive therapy. No recurrence was noted and the blood pressure returned to normal levels 4 months later. PMID- 20009421 TI - Evaluation of prognostic factors and establishment of a prognostic scoring system for canine primary immune-mediated hemolytic anemia. AB - Clinical courses of primary immune-mediated hemolytic anemia (pIMHA) in dogs are highly variable, however, limited information is available to predict their accurate prognoses. To evaluate the prognostic significance of clinical factors and to propose a scoring system to predict prognoses, the medical records of seventy-one dogs with pIMHA were reviewed. Overall mortality rate of dogs with pIMHA was 39% and most of the dogs died within 3 months from diagnosis. Sex, body weight, seasonality, packed corpuscular volume (PCV), platelet count (PLT), total plasma protein (TP), blood urea nitrogen, albumin, total bilirubin, sodium ion, prothrombin time, and fibrin/fibrinogen degradation products before immunosuppressive treatment can influence on survival time in dogs with pIMHA. A prognostic scoring system using a combination of sex, seasonality, PCV, PLT and TP can be statistically significant for raising the accuracy of prognostic prediction. Using the scoring system for prognostication in dogs with pIMHA may enable veterinarians to predict a prognosis easily and accurately. PMID- 20009422 TI - Chronic progressive polyarthritis in a female cat. AB - Feline chronic progressive polyarthritis is a rare immune-mediated disease that has only previously been reported in male cats. A one-year-old female cat was presented with anorexia, lassitude and lameness. The tarsal, carpal and elbow joints revealed swelling, pain, stiffness, crepitus and regional lymphadenopathy, and fever was present. The cat was clinically diagnosed with chronic progressive polyarthritis based on the fever, swelling of joints, imaging of erosive proliferative periosteal polyarthritis, positivity for antinuclear antibody, synovial fluid analyses and urinalyses. Both feline leukemia virus antigen and feline immunodeficiency virus antibody were positive. Using hair root DNA, polymerase chain reaction amplification targeting the sex-determining region on the Y chromosome gene amplified the fragment of DNA from a normal male cat, but not amplified from a normal female cat or the present cat. Accordingly, the present cat was classified as genetically female. Cyclosporine treatment was started, and the general condition and movement quickly improved and continued for 8 months post-diagnosis. This is the first report of chronic progressive polyarthritis in a female cat. PMID- 20009423 TI - Characterization of H5N1 highly pathogenic avian influenza virus isolated from a mountain hawk eagle in Japan. AB - On January 4, 2007, an emaciated mountain hawk-eagle was found in Kumamoto Prefecture, Japan. Highly pathogenic avian influenza (HPAI) virus subtype H5N1 was isolated from both tracheal and cloacal swabs of the dead bird. On January 13, an outbreak of HPAI, caused by H5N1 strain, occurred in a chicken farm in Miyazaki Prefecture. Within three weeks, three additional outbreaks had occurred (two in Miyazaki Prefecture and one in Okayama Prefecture). To investigate the relationship between the hawk-eagle isolate and chicken isolates, we studied the virus growth, pathogenicity, and phylogenetic information of this hawk-eagle isolate. The highest virus titer was found in the brain (10(7.25 )EID(50)/g), followed by trachea and muscle (10(2.65) and 10(2.50) EID(50)/g, respectively). Sequence analysis at the hemagglutinin (HA) cleavage site of this isolate revealed a typical virulent-type sequence, R-R-R-K-K-R. Phylogenetic analysis demonstrated that the hawk-eagle isolate belongs to Qinghai Lake type virus group. A homology search of the HA gene also showed major similarity (more than 99%) to the Miyazaki and Okayama isolates in 2007 and also Korean isolates in 2006. These results suggest that Qinghai Lake type H5N1 HPAI virus was newly introduced from Asian Continent into Japan, and had already present in natural environment of Kyusyu district in the beginning of January 2007. PMID- 20009424 TI - Two cases of bovine male pseudohermaphrodites with different endocrinological and pathological findings. AB - Two cases of bovine male pseudohermaphrodites (PH) were subjected for clinical investigation with transrectal ultrasonography, endocrinology with adoption of hCG-stimulation test, cytogenetics with analysis of sex chromosome and Y-specific DNA, and finally histological examination. Results were compared with normal calves. Case 1 was a 10-month-old calf with XX/XY chimeras, showing elevation of testosterone (T) levels, but no change in progesterone (P(4)) after hCG test, and possessed atrophied testes in the cavitas pelvis. Case 2 was an 18-month-old calf with SRY positive-XY chromosome, showing lower level of plasma T and P(4) after hCG test, and possessed atrophied testes and undifferentiated genital ducts. Both cases possessed female-like external genitalia with similar pathological findings, however endocrinological and cytogenetical aspects were different each other. PMID- 20009425 TI - Bacteriostatic activity of Whey Acidic Protein (WAP). AB - We have previously reported the action of whey acidic protein (WAP) inhibiting the proliferation of mouse mammary epithelial cells in the experiments utilizing in vivo and in vitro systems. We report herein the bacteriostatic activity of WAP. Western blot analysis demonstrated successful isolation of WAP from whey fractions of rat milk by column chromatography. The WAP fraction inhibited the growth of Staphylococcus aureus JCM2413 in a dose-dependent manner, but did not inhibit the growth of Escherichia coli. The bacteriostatic activity of WAP was highest at pH 6.6 and was not affected by the presence of 150 mM NaCl. A scanning electron micrograph of bacteria treated with WAP exhibited the disruption of the bacterial cell walls. PMID- 20009426 TI - Rectification of width and area of the ciliary cleft in dogs. AB - The width and area of the ciliary cleft (CCW and CCA) according to ultrasound biomicroscopy was rectified to compare dogs of different body sizes/weights, including toy breed (TBG), small breed (SBG), medium breed (MBG) and large/giant breed groups (LGBG). A linear correlation was detected between the natural log of body weight and the ocular axial length (OAL) reflecting the ocular size (OS; r=0.81, p<0.001). A significant positive correlation was found between OAL and the distance of the Schwalbe's line to the anterior lenticular capsule (SLD; r=0.87, p<0.001), suggesting that SLD could rectify canine OS. Rectified CCW (r CCW) and CCA (r-CCA) were calculated using SLD and fixed SLD as the SLD mean (SLDM). The SLDM was 2.55 mm in this study, and r-CCW and r-CCA were calculated as r-CCW = CCW x (2.55/SLD) and r-CCA = CCA x (2.55/SLD)(2). The CCW values of TBG, SBG and MBG were narrower than those of LGBG (p<0.05). There were significant statistical differences in CCA between all groups other than TBG and SBG (p<0.05). There were no significant differences in r-CCW and r-CCA in any of the groups. In addition, CCW/SLD did not show significant changes in dogs with different body sizes/weights. These results suggested that r-CCW, r-CCA and CCW/SLD can be used for comparison between dogs of different body sizes/weights. PMID- 20009427 TI - Increase in the neuraminidase activity of a nonpathogenic Newcastle disease virus isolate during passaging in chickens. AB - A pathogenic mutant of the Newcastle disease virus (NDV) was previously generated by passaging a non-pathogenic isolate from wild waterfowl. Velogenic mutant 9a5b (IVPI=2.67) contains three amino acid substitutions (128H, 495K and 573stop) in the hemagglutinin-neuraminidase (HN) protein, as compared with nonpathogenic waterfowl isolate 415/91 strain, and two of these (128H and 495K) were introduced after mesogenic 9a3b (IVPI=1.88). To investigate the role of the HN protein in NDV virulence, the function of HN protein such as neuraminidase (NA), Hemadsorption (HAd) and fusion promotion activities was examined by introducing the point mutations observed in passaged mutants into the HN gene cDNAs. In vitro functional assay using mutant protein expression demonstrated that the 128H substitution markedly increases NA activity and 573stop substitution increase NA and HAd activities. On the other hand, 495K substitution had little effect on any activities. These results indicate that a single amino acid substitution (128P to H) in the NDV HN protein affects the neuraminidase activity and is possibly correlated with the virulence. PMID- 20009428 TI - Muscarinic receptor subtypes mediating Ca2+ sensitization of intestinal smooth muscle contraction: studies with receptor knockout mice. AB - In the present study, we have characterized muscarinic receptor subtypes that mediate carbachol-induced Ca2+ sensitization of contraction in intestinal smooth muscle, using mutant mice lacking M(2) or M(3) muscarinic receptors or both receptor subtypes. In alpha-toxin-permeabilized muscle strips from wild-type (WT) mice, isometric tension responses to Ca2+ applied cumulatively (pCa 7.0-5.0) were increased when the muscarinic agonist carbachol (100 microM) was added to the medium, as judged from shifts of pCa-tension curves in both 50% effective concentration (EC(50)) and maximum response (E(max)) of pCa-tension curve. In preparations from M(2)-knockout (KO) mice, pCa-tension curves were also shifted by carbachol (100 microM), and the extents of the EC(50) and E(max) changes resembled those observed in preparations from WT mice. In preparations from M(3) KO or M(2)/M(3)-double KO mice, however, no significant changes in pCa-tension curves were obtained after carbachol application. The G(q/11)-type G-protein inhibitor YM-254890 (1 microM) completely blocked the Ca2+ sensitization of contraction induced by carbachol in M(2)-KO or WT preparations. The results strongly support the idea that the muscarinic activation of Ca2+ sensitization in intestinal smooth muscles is mediated by the M(3) muscarinic receptor coupled to G(q/11)-type G-proteins, without any significant involvement of the other muscarinic receptor subtypes including M(2). PMID- 20009429 TI - Establishment of vertebral heart scale in the growth period of the Japanese macaque (Macaca fuscata). AB - We performed a thoracic X-ray examination of 56 Japanese macaques to obtain normal reference values for vertebral heart scale (VHS). Mean VHS was 10.25 +/- 0.94 v. In males, mean VHS was 10.56 +/- 0.73 v, with no significant correlation to age or weight. In contrast, mean VHS in females was 9.97 +/- 1.03 v, and tended to decrease with increasing age and weight. These findings will facilitate the diagnosis of cardiac disease in Japanese macaques in the future. PMID- 20009430 TI - Effect of diet on the cardiovascular system in healthy beagles. AB - In management of canine cardiac disorders, cardiac drugs are commonly administered with food, although the cardiovascular effects of feeding and frequency of feeding are unknown. This study investigated the cardiovascular effect of feeding and frequency of feeding using a telemetry monitoring system in unanesthetized and unrestrained dogs. Systolic and diastolic blood pressure, heart rate and double product were steeply elevated during the five minutes immediately before feeding. They showed a sharp fall within 60 min after feeding time, especially the systolic and diastolic blood pressure, which showed lower values than from before feeding time. There were no significant differences in the means of preprandial elevation, postprandial fall or 24-hr averages in the once- and twice-daily feeding periods. PMID- 20009431 TI - Prevalence and molecular characterization of fluoroquinolone-resistant Escherichia coli isolated from diarrheic cattle in Korea. AB - A total of 176 E. coli isolates were retrieved from 203 diarrheic fecal samples collected from Korean cattle on 117 different farms. The most frequently observed resistance in E. coli isolates was to tetracycline (88.6%), followed by streptomycin (80.7%) and ampicillin (64.8%). Resistance to cefazolin, cefoperazone, cefepime and amikacin was very low. Of the 176 E. coli strains, forty (22.7%) isolates from 30 farms showed resistance to fluoroquinolones (FQ). All the FQ-resistant strains possessed double mutations at codons 83 and 87 in the gyrA gene, and a single mutation mostly at codon 80 in the parC gene, except in one isolate. The pulsed-field gel electrophoresis profiles of the FQ-resistant E. coli isolates were heterogeneous, but two or three isolates that showed an identical pattern originated from the same or different farms. This study demonstrates that FQ resistance is frequently observed in E. coli from diarrheic cattle and that mutations in the quinolone resistance-determining region are the same as those seen in E. coli originating from other animal species and humans. The FQ resistance in diarrheic cattle might have been mostly acquired independently, although the possibility of transmission of FQ-resistant E. coli within a farm or between farms is plausible. PMID- 20009432 TI - Gene expression profiling of diffuse large B-cell lymphoma -- comparison depending on the CD21 antigen expression. PMID- 20009433 TI - Fractionated administration of gemtuzumab ozogamicin for refractory acute myeloid leukemia. AB - It is difficult to decide an appropriate treatment strategy for elderly leukemia patients with other complications. We encountered 2 cases of refractory acute myeloid leukemia and safely treated the patients with fractionated administration of gemtuzumab ozogamicin (GO). Standard induction therapies were not effective for these patients. Moreover, they suffered from complications due to which their treatment options were restricted. Fractionated administration of GO (GO 3 mg/m(2) on days 1, 3 and 5) was accomplished safely and alleviated the patients' conditions. After treatment, these patients were followed by outpatient basis. We consider that this is an impressive treatment because fractionated administration of GO is potentially less toxic. Further, it will be helpful to maintain or improve the QOL of patients who are unable to receive intensive chemotherapy. These cases were significant because fractionated GO treatment is potentially less toxic and it will be helpful to maintain or improve the QOL of patients who can not receive intensive chemotherapy. PMID- 20009434 TI - Acute renal failure after high-dose methotrexate therapy in a patient with ileostomy. AB - High-dose methotrexate (HD-MTX) is an important treatment for Burkitt lymphoma, but can cause hepatic and renal toxicity when its clearance is delayed. We report a case of acute renal failure after HD-MTX therapy in a patient with ileostomy, The patient was a 3-year-old boy who had received a living-related liver transplantation for congenital biliary atresia. At day 833 after the transplantation, he was diagnosed with PTLD (post-transplantation lymphoproliferative disorder, Burkitt-type malignant lymphoma). During induction therapy, he suffered ileal perforation and ileostomy was performed. Subsequent HD MTX therapy caused acute renal failure that required continuous hemodialysis. We supposed that intravascular hypovolemia due to substantial drainage from the ileostoma caused acute prerenal failure. After recovery of his renal function, we could safely treat the patient with HD-MTX therapy by controlling drainage from ileostoma with total parenteral nutrition. PMID- 20009435 TI - Successful treatment with imatinib-combined chemotherapy for relapsed Philadelphia-positive acute lymphoblastic leukemia after allogeneic bone marrow transplantation. AB - The prognosis of patients with Philadelphia chromosome-positive acute lymphoblastic leukemia (Ph(+)ALL) relapsing after allogeneic hematopoietic stem cell transplantation is dismal. Here we describe a patient with post-transplant relapse of Ph(+)ALL, who has remained in complete remission (CR) for 30 months after relapse. A 55-year-old woman with Ph(+)ALL received allo-HSCT from an unrelated donor during first CR. The conditioning regimen consisted of fludarabine+melphalan, and graft-versus-host disease (GVHD) prophylaxis consisted of tacrolimus and short-term methotrexate. She achieved and maintained molecular remission without developing GVHD after transplantation, but suffered a hematologic relapse on day 871. She received imatinib-combined chemotherapy, and again achieved molecular remission. Since the completion of imatinib-combined chemotherapy, she has been receiving imatinib monotherapy. Although it has been reported that chemotherapy and imatinib are effective only transiently in patients with relapsed Ph(+)ALL, our patient has remained in molecular remission for 30 months after post-transplant relapse at the time of this report. Our case suggests that by continuing imatinib after the induction of molecular remission by imatinib-combined chemotherapy, the antileukemic activity of imatinib could achieve durable remission in combination with the graft-versus-leukemia effect. However, this needs to be investigated in studies involving a large number of patients. PMID- 20009436 TI - Multiple hepatocellular carcinomas developed 15 months after commencement of chemotherapy for elderly acute myelogenous leukemia. AB - In May 2006, a 72-year-old man with acute myelogenous leukemia (M4Eo) was admitted to our hospital. He had been receiving antiandrogen treatment for prostate cancer (after an operation in 1998) and treatment for diabetes mellitus. He received chemotherapy according to the JALSG GML200 protocol, which led to complete remission; however, in January 2007, his leukemia recurred. CAG combination chemotherapy also resulted in complete remission by May 2007. In August 2007, he developed multiple liver tumors, abdominal pain, and fever. Contrast-enhanced computed tomography revealed hypovascular tumors in both early and delayed phases. Angiography showed ring-like tumor staining and a massive tumor, similar to those seen in metastatic hepatocellular carcinomas (HCCs). He eventually died because of aggressive enlargement of liver tumors during the following month accompanied by the simultaneous recurrence of leukemia and unsuccessful embolization of the hepatic artery. Autopsy specimens showed fibrosis and considerable iron deposition in the liver, suggested secondary hemochromatosis due to transfusion. We also detected multiple moderately differentiated primary HCCs. Secondary hemochromatosis, androgen imbalance, and humoral factors from leukemic cells were believed to be the causes of the rapid onset and development of HCCs. PMID- 20009437 TI - POEMS syndrome complicated by follicular lymphoma. AB - A 59-year-old woman presented with ascites and intraperitoneal lymph node swelling. Pathological examination of the lymph node revealed follicular lymphoma. After a lymph node biopsy, she developed atypical genital bleeding, multiple endocrine disorders, polyneuropathy with a high plasma level of vascular endothelial growth factor (VEGF), and was diagnosed with POEMS syndrome. Following administration of methyl prednisolone, ascites immediately decreased and her performance status improved; however, about 18 months later, renal failure occurred, and she died despite increased steroid dosage. Lymph node swelling is often found in POEMS syndrome; however, its histological appearance is not well known, and it is very rare to be concomitant with malignant lymphoma. Therefore, it is important to perform a lymph node biopsy and investigate it in relation with VEGF. PMID- 20009438 TI - Hematopoietic recovery after administration of deferasirox for transfusional iron overload in a case of myelodysplastic syndrome. AB - Deferasirox (DFX) is a newly developed oral iron chelator that enables effective chelation with once daily administration. We describe here a case of transfusional-iron overloaded patient who experienced hematopoietic recovery after DFX administration. A 75-year-old woman with iron overload, who had been diagnosed with MDS (RCMD) and had received a transfusion of red blood cells and platelets regularly for 3 years, enrolled in the phase I clinical trial of ICL670 (DFX) in Japan. DFX administration steadily decreased her serum ferritin levels and chelated overloaded iron effectively. Interestingly, a year after initiation of the trial, she needed fewer blood transfusions, and no more transfusions after the 17th month of the trial. Even after suspending transfusions, her hemoglobin level and platelet count increased continuously, and she now has stable disease without blood transfusions. She has not received any specific treatment for MDS during this period. Examination of the bone marrow aspirates in the 35th month revealed dysplastic cells, indicating no remarkable change in the state of MDS. This case suggests that excess iron hampers hematopoiesis and that adequate iron chelation may improve hematological data in some iron-overloaded patients. PMID- 20009439 TI - Successful treatment with reduced-intensity stem cell transplantation for secondary myelofibrosis following polycythemia vera. AB - Ten years after being diagnosed with polycythemia vera, a 55-year-old woman required frequent blood transfusion due to secondary myelofibrosis. She underwent reduced-intensity stem cell transplantation (RIST) from an HLA-identical sibling donor. Since mixed chimerae were identified in the peripheral blood at day 35, cyclosporine was withdrawn. At day 73, she developed acute graft-versus-host disease of the liver, while simultaneous resolution of splenomegaly occurred and complete donor chimerism in the peripheral blood was achieved. Frequent red blood cell transfusion was required until day 300 after transplantation. Thus, RIST for an older patient with secondary myelofibrosis was successful without severe treatment-related morbidity. This case suggests that RIST could be an effective treatment modality for secondary myelofibrosis. PMID- 20009440 TI - Multiple myeloma with multiple and bulky extramedullary plasmacytomas as initial presentation. AB - We herein describe a rare case of multiple myeloma with an aggressive clinical course and the unusual manifestation of multiple organ involvement by plasma cells. A 58-year-old man noted difficulty in walking due to progressive swelling of his left lower limb. CT scan revealed a huge mass in the inguinal region in addition to masses located on the head, and in the aero-digestive tract and spinal canal. The pathological diagnosis of plasmacytoma was made on biopsied specimens of these masses, while plasma cells did not increase (5.8%) in aspirated bone marrow obtained at the same time. Serum IgG level was 6,387 mg/dl and immunoelectrophoresis demonstrated monoclonal IgG-kappa in the serum. Chemotherapy with vincristine, adriamycin, dexamethasone, subsequent high-dose cyclophosphamide, and irradiation involving both thoracic vertebral canal and inguinal regions resulted in improvement of initial symptoms. However, the patient relapsed soon after; new lesions developed in various parts of the body, including the left thigh and body trunk. Salvage therapy including bortezomib was no longer effective, and he eventually died 10 months after the initial diagnosis. Autopsy revealed the diffuse involvement of plasma cells of multiple organs, including the liver, spleen, abdominal lymph node, and bone marrow in addition to the left leg. PMID- 20009441 TI - Successful treatment with autologous peripheral blood stem cell transplantation for acquired immunodeficiency syndrome (AIDS)-related malignant lymphoma. AB - A 62-year-old man was diagnosed with human immunodeficiency virus (HIV) infection while suffering from recurrent herpes zoster infection. Laboratory examination revealed CD4(+) lymphocyte count 16 cells/mul and HIV loading 150,000 copies/ml at presentation. In addition, he had multiple lymph node swelling. Histologic diagnosis of a biopsied lymph node was diffuse, large, B cell-type malignant lymphoma. The karyotype of the lymphoma cells was t(8;14)(q24;q32), which was confirmed by G-banding and fluorescent in situ hybridization. Positron emission tomography (PET)-combined CT scanning revealed systemic extranodal tumors involving the gastrointestinal tract, pancreas, and bone marrow. The clinical stage of the lymphoma was IVB and the international prognosis index was categorized as high. Complete remission (CR) of the lymphoma was obtained after 2 courses of CHOP (cyclophosphamide, adriamycin, vincristine, prednisolone) chemotherapy and 4 subsequent courses of rituximab-combined CHOP (R-CHOP). Highly active antiretroviral therapy (HAART) was started at the initiation of CHOP. Because of the poor prognosis of AIDS-related lymphoma, he received autologous peripheral blood stem cell transplantation with the MEAM protocol (ranimustine, etoposide, cytarabine, melphalan) as a conditioning procedure without a severe infectious episode. He remains in CR 24 months after the transplantation. PMID- 20009442 TI - Effective and well tolerated treatment with melphalan and dexamethasone for primary systemic AL amyloidosis with cardiac involvement. AB - A 60-year-old woman was admitted with acute heart failure and was diagnosed as having primary systemic AL amyloidosis with cardiac involvement by endomyocardial biopsy. Electrophoresis revealed an IgG-lambda monoclonal component and amyloidosis was evident in the gastric and rectal mucosa. Her cardiac function at diagnosis was poor, including an ejection fraction of 59% and IVS of 19 mm, and serum cardiac troponin T (cTnT) was elevated (0.12 ng/ml). She was treated with melphalan-dexamethasone (Mel-Dex) therapy once a month. After more than a year, cardiac function and performance status were maintained, with decreasing levels of cTnT, indicating that Mel-Dex represents a feasible and effective therapeutic option for patients with AL amyloidosis with cardiac dysfunction. PMID- 20009443 TI - Thrombotic thrombocytopenic purpura presenting with reversible posterior leukoencephalopathy syndrome. AB - A 48-year-old man presented with consciousness disturbance with vasogenic edema in the occipital lobe on brain CT. The diagnosis of reversible posterior leukoencephalopathy syndrome (RPLS) was made. His hypertension was refractory to treatment, and his neurological disabilities and CT abnormalities, along with renal dysfunction, became worse. Hemodialysis and strict management of blood pressure resolved the neurological findings and the lesions on brain CT. However, one week later, consciousness disturbance and brain CT abnormalities recurred. At that time, hemolytic anemia with fragmented erythrocytes, thrombocytopenia, and renal dysfunction became apparent. We therefore diagnosed thrombotic thrombocytopenic purpura (TTP). Plasma exchange and vincristine administration improved not only the clinical findings of TTP, but also consciousness disturbance and brain CT abnormalities. We concluded that latent TTP had caused RPLS in this patient. PMID- 20009444 TI - Iron deficiency anemia successfully treated by Helicobacter pylori eradication in a patient with idiopathic thrombocytopenic purpura. AB - A 53-year-old woman had demonstrated idiopathic thrombocytopenic purpura (ITP) and iron deficiency anemia (IDA) since 1978. Although she was treated with prednisolone for ITP and oral iron compounds for IDA, neither ITP nor IDA showed any improvement. Since her (13)C-urea breath test was positive, Helicobacter pylori (H. pylori) eradication therapy was performed in 2001. The therapy was effective for IDA but not for ITP. Analysis of cases such as this will be useful for clarifying the mechanisms underlying the development of ITP and IDA associated with H. pylori. PMID- 20009445 TI - Population drinking and fatal injuries in Eastern Europe: a time-series analysis of six countries. AB - AIMS: To estimate to what extent injury mortality rates in 6 Eastern European countries are affected by changes in population drinking during the post-war period. DATA AND METHODS: The analysis included injury mortality rates and per capita alcohol consumption in Russia, Belarus, Poland, Hungary, Bulgaria and the former Czechoslovakia. Total population and gender-specific models were estimated using auto regressive integrated moving average time-series modelling. RESULTS: The estimates for the total population were generally positive and significant. For Russia and Belarus, a 1-litre increase in per capita consumption was associated with an increase in injury mortality of 7.5 and 5.5 per 100,000 inhabitants, respectively. The estimates for the remaining countries ranged between 1.4 and 2.0. The gender-specific estimates displayed national variations similar to the total population estimates although the estimates for males were higher than for females in all countries. CONCLUSIONS: The results suggest that changes in per capita consumption have a significant impact on injury mortality in these countries, but the strength of the association tends to be stronger in countries where intoxication-oriented drinking is more common. PMID- 20009446 TI - [New response evaluation criteria in solid tumours-revised RECIST guideline (version 1.1)]. AB - This paper is an overview of the new response evaluation criteria in solid tumours: revised RECIST guideline (version 1. 1), with a focus on updated contents. PMID- 20009447 TI - [The process of the revision of staging system for lung cancer(UICC-7)]. AB - The agenda for a new staging system for lung cancer has been determined and proposed to the UICC and AJCC by the IASLC (International Association for the Study of Lung Cancer) staging committee. Publication by both organizations is scheduled late this year. For the present revision, the IASLC launched a staging project early and accumulated data on more than 100, 000 patients from all continents. Based on these data, an agenda was established, in which the new categorization of T according to tumor diameter and satellite nodules, the creation of new lymph node chart, and the new stage grouping are characterized. The new staging system (UICC-7) is scheduled to take effect as of from January, 2010. PMID- 20009448 TI - [Clinical utility of PET-CT for esophageal cancer]. AB - It is important to diagnose the exact clinical staging according to the improvement of treatment for esophageal cancer. PET examinations for esophageal cancer are now covered by Japanese government health insurance, so the number of PET scans is now gradually increased in many institutions and their utility has been reported. In this report, we reviewed the conventional diagnostic tools for esophageal cancer and evaluated the clinical significance of PET-scans conventionally associated with it. The depth of tumor invasion of a primary tumor is detectable in more than 80% of cases of SM2 or deeper invasion. With lymph node metastasis, the sensitivity was 66. 7% and specificity 93. 5%. With distant metastasis, there is reportedly a high detection rate in bone metastasis and liver metastasis. However, with lung metastasis, reference to CT scans rather than PETscans is important in making a carefull diagnosis. In judging the effect of preoperative chemoradiotherapy, PET-CT was suggested to be an effective diagnostic tool. In addition, PET may be a useful diagnostic tool for postoperative follow-up patients. Finally, further investigation may well be necessary in the future. PMID- 20009449 TI - [Clinical usefulness of FDG-PET for pancreatic cancer]. AB - PURPOSE: The purpose of this study was to estimate the usefulness of positron emission tomography (PET) in dividing the uptake with and without the main tumor for the treatment of patients with pancreatic cancer. METHODS: Ninety-eight patients with primary pancreatic cancer were evaluated with 18F-FDG-PET. For the main tumor, the maximum standardized uptake values(SUVmax)were compared with clinicopathological factors and analyzed. We examined the site of accumulation and the rate of malignancy without the main tumor. RESULTS: For the accumulation of FDG in the main tumor, the high SUVmax level was significantly correlated with T-category in TNM classification (p=0. 003), tumor invasive size (>3 cm) (p <0. 001), CA19-9 levels>100 U/mL) (p=0. 002). The overall survival of the group in which SUVmax was less than 7. 5 was better than that of the group in which it was more than 7. 5 (p=0. 03). Meanwhile, 58 patients (59%) showed the accumulation of FDG except for the main tumor. Lymph node uptake was shown in 44% of them. As for visceral accumulation, the liver was 11, lung 10, pancreas except main tumor 9, thyroid 7, peritoneal wall 3, colon 2, gall bladder 2, and bone 1. As for the rate of malignancy among them, the liver was 100%, lung 50%, pancreas except main tumor 0%, thyroid 29%, peritoneal wall 67%, colon 50%, gall bladder 50%, and bone 0%. CONCLUSIONS: We conclude that FDG-PET is a useful tool for predicting the prognosis in pancreatic cancer, and for detection of distant metastases and hidden malignant disease. FDG-PET has an important clinical impact on the selection of proper treatment. PMID- 20009450 TI - [Roles of positron emission tomography/computed tomography (PET/CT) in the management of colorectal cancer]. AB - In diagnosis and treatment of colorectal cancer, PET/CT has high sensitivity and specificity in comparison to other modalities, and in the near future is expected to play important roles in these areas. However, because of the high cost, PET/CT must be used cost-effectively. In the diagnosis of colorectal cancer with PET/CT, evaluation of cancer growth is possible, but evaluation of tumor invasion is inadequate. In the diagnosis of lymph node metastases, PET/CT provides higher sensitivity of evaluation in distant lymph nodes. The reason for difficulty in proximal lymph node evaluation is due to the close proximity of the main cancer tumor. In the diagnosis of liver metastases, meta-analysis indicates the need for higher sensitivity and specificity compared to CT and MRI. In the diagnosis of pulmonary metastases, chest CT should be obtained in very early pulmonary metastases because of the tiny tumor size in the early stage. In the diagnosis of local recurrence, discrimination between postoperative change and recurrence is difficult, but PET/CT is very useful as a qualitative diagnostic tool. Recent reports have also indicated the usefulness of PET/CT in the evaluation of treatment efficacy in chemotherapy or radiotherapy. PMID- 20009451 TI - [The role of / 18FDG PET/CT for the initial staging and therapy in primary breast cancer]. AB - PURPOSE: To evaluate the utility of / 18F-FDG PET/CT in the initial staging and predicting response to neoadjuvant chemotherapy ( NAC) for primary breast cancer. METHODS: One hundred fourteen patients with primary breast cancer were preoperatively evaluated with PET/CT. The diagnostic value of PET/CT was compared in the final histopathology following surgery. Twenty patients also underwent PET/CT at baseline and after 4 and 8 courses of NAC. Changes (deltaSUV)in the maximum standard uptake value (SUVmax) between baseline study and after 4 or 8 courses of NAC were compared with the pathological response. RESULTS: In the detection of primary breast cancer, PET/CT was 80. 7% sensitive, and especially, less sensitive in the detection of tumor size less than 10mm (66. 7%), low tumor grade (61. 3%), and ductal carcinoma in situ (50%). For axillary staging, PET/CT was 51. 9% sensitive and 95. 3% specific; in particular, it was less sensitive for small (<10mm) axillary lymph nodes (29. 4%) and could not detect micrometastasis. A cut-off deltaSUV value of 91. 9% was determined between baseline and after 4 courses of NAC differentiated responders(pathological CR)and non-responders, with a sensitivity of 80%, and a specificity of 78. 6% (p=0. 038). CONCLUSION: Although PET/CT may have limited diagnostic value for detecting primary tumor and axillary staging, it is a valuable modality for monitoring the response to therapy. PMID- 20009452 TI - [Malignant lymphoma]. AB - 18-Fluoro-deoxyglucose positron emission tomography (FDG-PET) has become widely used in the management of patients with malignant lymphomas. FDG-PET has been evaluated in pretreatment staging, restaging, monitoring during therapy, and posttherapy surveillance. The Ann Arbor staging system was initially based on physical examination and bone marrow evaluation, but more recently, CT scans or 67gallium scintigraphy have been incorporated. FDG-PET may provide complementary information to conventional staging methods, and may be of particular value prior to therapy for patients who appear to have stage I or II disease and for whom radiation therapy is being considered. FDG-PET has technical limitations, variability of FDG avidity among different lymphoma histologic subtypes, and in a large number of etiologies shows false-negative and false positive results. Most studies of FDG-PET involve patients with Hodgkin's disease or diffuse large B cell lymphoma. FDG PET in lymphoma is being incorporated into the response assessment in lymphoma as published by the Imaging Subcommittee of International Harmonization Project in Lymphoma. New guidelines, the Revised Response Criteria for Malignant Lymphoma, are presented incorporating PET, IHC, and flow cytometry for definitions of response in non-Hodgkin's and Hodgkin's lymphoma. They should reduce the variability among studies. Standardized definitions of end points are provided. Standardized FDG-PET will hopefully lead to improved outcome for patients. PET as a biomarker has the potential to change the current model of drug development. PMID- 20009453 TI - [Application of PET in radiation oncology]. AB - Positron emission tomography (PET), especially F-18 fluorodeoxyglucose (FDG)-PET, has been recently used to verify the target volume in radiation treatment planning (RTP) for malignancies. The utility of FDG-PET/CT in defining gross tumor volume (GTV) has been shown in many studies, and the target delineation by a fixed threshold of the maximum standardized uptake value (40-50%) is suggested to be useful in RTP for lung cancer, head and neck cancer, etc. But, the spatial resolution, sensitivity, and specificity of PET are not always enough to define the difference between the GTV and the clinical target volume(CTV). Furthermore, FDG-PET is frequently used in the clinical staging before the treatment, and is also applied to the response evaluation after the treatment. This review focuses on the developing applications of PET in radiation oncology. PMID- 20009454 TI - [The role and limitations of FDG-PET in head and neck cancer]. AB - 18F-fluorodeoxyglucose positron emission tomography (FDG-PET) is a nuclear medicine imaging technique which has been increasingly applied as a diagnostic tool. The usefulness of FDG-PET may be described as follows: I . Early detection of cancer; II. Diagnosis of cancer; III. Detection of nodal or distant metastasis, and double cancer; IV. Detection of unknown primary tumor with metastatic neck lesions; and V. Evaluation of treatment of head and neck cancer. FDG-PET is especially useful to detect distant metastasis, double cancer with head and neck cancer and unknown primary tumor with metastatic neck lymph nodes. The conventional modalities, e. g., CT or MRI, show anatomical images in the body. On the other hand, FDG-PET reveals three-dimensional images of functional processes of the glucose metabolism. FDG-PET can estimate metabolic activity in cancer and is useful in evaluating or monitoring the response to concurrent chemoradiotherapy of the head and neck cancer. However, we should recognize the limitations of FDG -PET. An acute inflammatory disease shows high FDG uptake like cancer. It is difficult to detect early-stage esophageal cancer or to diagnose parotid gland cancer. PMID- 20009455 TI - [The effectiveness of concomitant radiotherapy with S-1 and nedaplatin for tongue cancer]. AB - The development of reconstructive surgery and the use of free flaps have allowed for a larger dissection range even for advanced tongue cancer, resulting in an improvement of the prognosis. However, both the postoperative swallowing and masticatory function are still considered to have not yet reached a satisfactory level. Accordingly, our department has been administering concurrent chemoradiotherapy (CCRT) for advanced cancer to preserve the organ and the function; there are cases in which even comparatively small tumors are difficult to dissect due to the occurrence site. We have been treating these cases using CCRT as well. We herein report our results of 10 tongue cancer cases in which CCRT with S-1 and Nedaplatin (hereinafter, referred to as SN therapy) was administered in our department from April 2002 to October 2008. The complete response rate of the SN therapy was 60. 0% (6 of 10 examples). The 5-year disease specific survival rates were 50. 0% for Stage II, 75. 0% for Stage III, and 75. 0% for Stage IV, respectively. PMID- 20009456 TI - [Analysis of factors associated with quality of life for older adult patients with non-small cell lung cancer receiving outpatient gemcitabine therapy as an alternative inpatient therapy]. AB - An increasing number of patients with lung cancer are undergoing outpatient chemotherapy as an alternative to inpatient therapy. To investigate the factors influencing the quality of life (QOL) for these patients, we administered a QOL questionnaire, The QOL Questionnaire for Cancer Patients Treated with Anticancer Drugs (QOL-ACD) to 8 older adult patients with non-small cell lung cancer. The medical factors related to the overall QOL scores and other categories indicating activity, physical condition, psychological condition, social relationship, psychological condition and face scale were analyzed. No significant decrease in each factor was observed in this study. PMID- 20009457 TI - [Growth rates of gastric cancer in the aged]. AB - Yamashiro et al reported "growth patterns of gastric cancers in the aged" in 1991 using 219 autopsy cases and 33 followup cases of gastric cancer in the natural course without operations obtained for 16 years. They also showed a certain curve of cancer area and ordinal number of tumor size on the logarithmic scale of both axes obtained from 120 gastric cancer cases from autopsy. We considered the curve by Yamashiro as a model curve of cancer growth by regarding ordinal numbers of tumor size as course times of tumor growth. Then we re-examined the 33 follow-up cases and calculated the tumor growth rate in each case by means of comparing case data with the model curve described above. In the result, the follow-up cases were grossly divided into 5 groups of ratios of growth rate 2(0), 2(1), 2(2), 2(3) and 2(4). In some articles about the cancer growth-rate of digestive organs, their data-values of doubling-time are suggestive of an arrangement with groups and gaps in it, respectively. They seem supportive of our results above. PMID- 20009458 TI - [Hepatic resection after neoadjuvant therapy for initially unresectable colorectal liver metastases]. AB - BACKGROUND: Recently, an increased number of reports have been published on liver resection following neoadjuvant chemotherapy ( NAC) in patients with initially unresectable colorectal liver metastases (IUCLM). However, the definition of unresectable liver metastases differs among institutions. The size of liver tumor B5 cm and number of tumors B5 is commonly a contraindication for resection of liver metastases. The present study was performed to compare the short and longterm results between patients who underwent liver resection following NAC for IUCLM and those with multiple bilobar metastases for initially resectable liver metastases. METHODS: Twenty-seven patients with multiple bilobar liver metastases between 1994 and 2007 were divided into two groups, i. e. 11 patients who underwent liver resection following NAC for IUCLM and 16 patients who initially underwent liver resection. NAC was used in three in J-IFL and eight cases in mFOLFOX6. RESULTS: All eleven patients with IUCLM were H3/grade C. The median course of NAC was 6 (4-6 courses, Mean+/-SD: 6+/-2 courses). The objective overall response rate was 100% (11/11). H3 of eleven patients was changed to two in H1 and nine in H2 after chemotherapy. Grade C of 11 patients was down-staged in 4 in grade Band 2 in grade A. The H factors and grade of 16 patients who initially underwent liver resection were H16H28H32 and grade A4/B6/C6, respectively. The disease-free and overall survival after resection of colorectal liver metastases between patients with initially unresectable and resectable liver metastases were not significantly different. CONCLUSIONS: NAC enables liver resection in some patients with IUCLM. It should be performed not only preoperatively but also postoperatively for IUCLM because of better survival after surgery. PMID- 20009459 TI - [Usefulness of clinical study of irinotecan and cisplatin for recurrent ovarian cancer]. AB - So far, there is no recommended evidence-based chemotherapeutic regimen for recurrent or refractory ovarian carcinoma in the guideline. In our institute, combination chemotherapy of CPT-11 and CDDP (CPT-P) was administered for recurrent or refractory ovarian carcinoma patients previously treated with regimens including taxane. In fourteen cases treated with CPT-P, there were 2 cases of CR, 2 of PR, 3 of SD, and 7 of PD. Overall response rate was 28. 5%, and the median progression-free interval was 5.5 months. No severe side effect was observed except in one patient in whom chemotherapy was discontinued due to grade 4 neutropenia and grade 3 diarrhea. Thus CPT-P seemed to be effective and safe for the recurrent or refractory ovarian carcinoma previously treated with taxane included regimen. PMID- 20009460 TI - [Clinical investigation of bisphosphonate-related osteonecrosis of the jaws]. AB - We examined the clinical features of bisphosphonate(BP)-related osteonecrosis of the jaws(BRONJ), a serious complication resulting from intravenous BP treatment for multiple myeloma and malignant tumors with bone metastasis. We retrospectively reviewed the medical records of 36 patients who received intravenous BP therapy for the above-mentioned conditions, at Sapporo Medical University Hospital between July 2006 and October 2008. BP therapy caused BRONJ in 7 of 24 patients, but did not affect the bones of the other 17 patients. The other 12 of the 36 patients involved in the study were prescribed BP only after they had undergone an oral examination and treatment for dental inflammation. Of these patients, 7 developed BRONJ with BP treatment, after tooth extraction or acute dental inflammation. Treating dental inflammation before prescribing BP prevented the development of BRONJ. BRONJ is highly intractable and does not resolve with the standard treatment for osteomyelitis. Therefore, preventive therapy, which can be achieved by cooperation between medical doctors and dentists, is currently the most effective strategy for BRONJ. Conservative treatment with antibiotics may also be useful for maintaining or improving the quality of life of BRONJ patients. PMID- 20009461 TI - [Clinical evaluation of calculating carboplatin doses using modification of diet in renal disease (MDRD) estimate and adverse events]. AB - A modified diet in renal disease (MDRD), a formula to estimate glomerular filtration rate (GFR), was proposed by Levey in 2006. In this study, we compared the dosage of carboplatin (CBDCA) calculated using MDRD with that calculated by conservative creatinine clearance (Ccr), and investigated the actual dosage given and the incidence of its adverse effects. In the 101 patients undergoing chemotherapy including CBDCA, the dosage calculated from the estimated GFR was 16% lower than that based on the estimated Ccr. This difference was greater in those under 65 years, women and those with a body mass index (BMI) > or =25. The most prominent incidence of adverse effects was thrombocytopenia in patients with lung cancer. In men, a serum creatinine level of > or =0. 6 mg/dL, GFR of <50 mL/min/1. 73 m(2) and a combined use of gemcitabine were cited as the factors responsible for the development of thrombocytopenia. It was concluded that the MDRD formula is an effective tool for evaluating patients with impaired renal function. It was suggested, on the other hand, that the dosage for medication should be decided by giving due consideration to factors other than renal functions. PMID- 20009462 TI - [Cost-effective analysis of rotation from sustained-release morphine tablet to transdermal fentanyl of matrix type or sustained-release oxycodone tablet]. AB - The present study was undertaken to determine the pharmacoeconomics of switching from sustained-release morphine tablet to matrix type (MT) of transdermal fontanel or sustained-release Oxycodone tablet. Cost-effective analysis was performed using a simulation model along with decision analysis. The analysis was done from the payer's perspective. The cost-effective ratio/patient of transdermal MT fontanel (22, 539 yen)was lower than that of sustained -release Oxycodone tablet (23, 630 yen), although a sensitivity analysis could not indicate that this result was reliable. These results suggest the possibility that transdermal MT fontanel was much less expensive than a sustained-release Oxycodone tablet. PMID- 20009463 TI - [Successful gefitinib every other day administration in an advanced lung cancer patient with brain metastasis after gamma knife radiotherapy and chemotherapy]. AB - A 69-year-old female, who had been admitted to another hospital with a complaint of headache and there detected brain tumor, was referred to our hospital for further examination and therapy. The patient was diagnosed as having advanced lung cancer with multiple brain metastasis. She was treated with five courses of a combination of chemotherapy consisting of carboplatin and paclitaxel following gamma knife radiotherapy. She showed a remarkable response, however, she experienced the side effects of general fatigue and numbness in her extremities, which were intolerable. Then, therapy with gefitinib alone was chosen as second line chemotherapy. After one month, this therapy was discontinued due to grade 3 skin trouble and rash. When her condition improved, every other day oral administration of gefitinib was resumed. She has been treated on an outpatient basis because of no severe adverse reactions. The patient is alive with good performance status (PS) 0 more than one year after taking gefitinib. Furthermore, the primary and metastatic tumors are not enlarged. Gefitinib orally administered every other day could be a promising regimen as second-line chemotherapy for patients with lung cancer and brain metastasis. PMID- 20009464 TI - [Complete response by S-1 with multiple pulmonary metastases 4 years after lung resection-a case report]. AB - A male patient in his seventies presented at our hospital with a chief complaint of an abnormal shadow on a chest scout film. We suspected non-small cell carcinoma of the right upper lobe on computed tomography (CT). The patient underwent partial resection of the right upper lobe by video-assisted thoracoscopic surgery in May 2004, frozen sections of which showed poorly differentiated squamous cell carcinoma, consistent with the tumor size 2.5 x 1.7 cm. We performed right upper lobectomy and lymphadenectomy. Final pTNM was T1N0M0, stage IA. The patient had been followed every three months until a coin lesion appeared in the right lower lobe in April 2007. He chose radiation therapy of 48 Gy in total at another hospital with the result that the lesion disappeared. However, the disease recurred in May 2008. Bilateral multiple lung metastases and mediastinal lymph nodes involvement were confirmed by CT. Two courses of docetaxel 60 mg/m(2) (90 mg/body) and cisplatin 80 mg/m / 2 (120 mg/body) were not effective with adverse reaction grade 3. Less docetaxel and cisplatin resulted in a grade 4 reaction. The regimen was changed to S-1 60 mg per day with three weeks on and two weeks off. After two courses complete remission was confirmed at a chest X-ray examination. He has been free of disease with tegafur/uracil 600 mg per day for 4 months. PMID- 20009465 TI - [A case of meningeal carcinomatosis of lung adenocarcinoma well controlled by re treatment with gefitinib]. AB - A 74-year-old-woman who had never smoked was diagnosed in 2004 with cT3N2M1, stage IV primary pulmonary adenocarcinoma. After seven courses of chemotherapy with carboplatin and paclitaxel, she was given gefitinib as second-line therapy and made satisfactory progress. However, gefitinib was discontinued after 3 years of treatment due to re-growth of the tumors. She was then given chemotherapy with docetaxel as a third-line therapy. Over the course of time, meningeal carcinomatosis occurred in conjunction with her previous disease. Upon re treatment with gefitinib, her meningeal carcinomatosis showed some improvement despite the growing primary tumor, so her QOL was improved. She is now visiting a hospital as an outpatient. When analysis of the EGFR gene mutant was conducted, deletion mutation of E746-A750 in exon 19 was revealed in the 2004 pulmonary tissue, as well as the cytological examination of cerebrospinal fluid and pulmonary tissue after recurrence. No change has been observed. Once gefitinib proved effective, re-treatment with gefitinib was considered useful after its discontinuation. PMID- 20009466 TI - [A case of aggressive primary squamous cell carcinoma of the breast refractory to preoperative neoadjuvant chemotherapy]. AB - A 42-year-old woman came to our hospital complaining of an enlarging tumor in her right breast. A core needle biopsy of the involved area demonstrated primary squamous cell carcinoma (SCC) of the breast. Preoperative neoadjuvant chemotherapies included 5-fluorouracil: 5-FU+epirubicin: EPI+cyclophosphamide: CPA (FEC), paclitaxel (PTX), vinorelbine (VNR) and 5-FU+nedaplatin (254-S) given in the order listed, but the mass continued to enlarge. Therefore, surgical resection was performed. Local recurrence and lung metastasis occurred after surgery. Although postoperative adjuvant chemotherapy consisting of carboplatin (CBDCA) +etoposide (VP-16) was administered, the patient died three months after surgery. SCC of the breast is generally treated according to clinical stage using protocol for common types of breast cancer. However, effective regimens have not been established for SCC, because it tends to be treatment-refractory. Therefore, in patients with SCC, it is important to consider surgery at an earlier stage than would be considered for a common breast cancer requiring preoperative neoadjuvant chemotherapy. PMID- 20009467 TI - [A case of recurrent breast cancer with extensive liver metastasis successfully treated with endocrine therapy]. AB - A 5 6-year-old woman, who underwent breast-conserving surgery and radiation (60 Gy) therapy in July, 1992, at the age of 40, was diagnosed with pT1aN0M0, pStage I. She was administered tamoxifen (TAM) as adjuvant therapy. However, she underwent microdochectomy for DCIS in her contralateral breast in June, 1998. TAM was given till August, 1999. In June, 2006, at the age of 54, 14 years after initial surgery, CT revealed extensive liver masses which were diagnosed as liver metastasis by liver biopsy. Receptor status was positive for ER and PgR, and negative for HER2. AC was started as a first-line chemotherapy ( 4 courses), but did not prove effective. She refused second-line chemotherapy, so letrozole was selected, and subsequently resulted in PR of the liver metastasis. However, 8 months later, with a liver metastasis relapse, exemestane followed by tamoxifen, medroxyprogesterone acetate, and high-dose toremifene were administered sequentially, resulting in long-time disease control. In conclusion, endocrine therapy might be an effective option even in a visceral crisis, if metastatic tumors have showed slow growth and there is positive hormone receptor status. PMID- 20009468 TI - [A case of advanced breast cancer with multiple bone metastases responding to docetaxel and high-dose toremifene as fourth-line chemo-endocrine therapy]. AB - A 55-year old woman, who underwent left mastectomy (Bt+Ax), was revealed to have sternum metastasis by postoperative 99mTc bone scanning(T1bN1M1). She received daily aromatase inhibitor (anastrozole), as a primary systemic endocrine therapy, and biweekly pamidronate for metastatic breast cancer. However, she depended on folk medicine a year later, at which time the primary treatment was discontinued. Another year later, the bone metastases developed with increased serum levels of tumor markers (CEA, CA19-9, and NCC-ST-439). Then, she underwent three different regimens of systemic chemo-endocrine therapy over the following three years, including CAF+MPA as the first-line, paclitaxel (PTX) + anastrozole as the second line, and S-1+anastrozole as the third-line regimen. She recently completed 10 courses of the fourth-line regimen[tri-weekly docetaxel (DOC) and high-dose toremifene (TOR 120 mg/day)], which reduced levels of 99mTc accumulation in the multiple bone metastases and levels of the serum tumor markers to the normal range. No severe adverse events occurred except peripheral thrombovasculitis (grade 2) in her left anterior arm during the fourth regimen. She recently maintains the current status by taking a regular dose (40 mg/day) of toremifene for 5 months. Combination treatment with DOC and high-dose TOR can be one of the worthwhile regimens as systemic chemo-endocrine therapy for patients with advanced breast cancer who develop bone metastases. PMID- 20009469 TI - [A case of recurrent breast cancer responding to vinorelbine/trastuzumab combination therapy]. AB - A 60-year-old woman with Stage II, ER-positive, PgR-positive, HER2 (2+) cancer in the right breast underwent right mastectomy with right axillary dissection after chemotherapy with EC followed by docetaxel (DOC) alone. Exemestane was used for postoperative adjuvant treatment. She underwent a right chest wall tumor resection for local recurrence. Hormone therapy was continued with toremifene in place of exemestane. In December 2007, two years after the second surgery, CEA was elevated and PET showed a local recurrence in the right chest wall and metastases to the right axillary nodes and liver. The tumor was ER-positive, PgR negative and HER2 (3+) at recurrence, and vinorelbine/trastuzumab combination was initiated as first-line chemotherapy for the recurrent lesion and liver metastasis. All lesions in the right chest wall, right axillary nodes and liver disappeared from PET and CT images after five courses of the regimen, resulting in clinical CR. Vinorelbine combined with trastuzumab appears to be a useful therapy for HER2-positive recurrent breast cancer. PMID- 20009470 TI - [A case report of complete response by S-1/docetaxel combination chemotherapy for advanced gastric cancer]. AB - A 52-year-old male patient with advanced gastric cancer and multiple lymph node metastasis was treated by S-1/docetaxel combination chemotherapy from April, 2004. As of January 2005, after the tenth course, a scarred tumor was observed with an endoscope, and negative neoplastic cells were found on endoscopic biopsy. After the sixteenth course in July, 2005, CT scan showed that the lymph node metastasis had disappeared. The primary and metastatic lesions were regarded to have responded completely. A total of 26 courses of chemotherapies were undergone until March, 2006. We are taking a wait and see approach at this writing without treatment since then. The patient has had a recurrence-free survival for about five years since the onset. The results suggested that the S-1/docetaxel chemotherapy for metastatic gastric cancer had confirmed feasibility and sufficient efficacy while maintaining the patient's quality of life. PMID- 20009471 TI - [A Case of gastric endocrine cell carcinoma successfully treated by FU plus irinotecan(CPT-11)adjuvant therapy against recurrent metastases]. AB - A case of gastric endocrine cell carcinoma successfully treated by FU (5-FU/UFT) +irinotecan (CPT-11) adjuvant therapy against recurrent metastases is reported with some discussion. A 69-year-old man was admitted to our hospital with severe anemia. He was diagnosed with advanced gastric cancer, T3N1H0P0M0, Stage IIIa. Total gastrectomy with pancreato-splenectomy with D2 lymph node dissection was done for curative resection. The pathological diagnosis was gastric endocrine cell carcinoma because Grimelius and Chromogranin A stained positive histologically. Seven months after operation, recurrent liver metastases with tumor embolism of the portal vein were revealed by enhanced CT examination. FU (5 FU/UFT) +CPT-11 was done as the first-line adjuvant chemotherapy. Metastatic lesion of the liver and portal vein tumor embolism was decreased. Tumor marker CA19-9 level was also decreased and within normal limits. This therapy was evaluated as a partial response (PR) in twelve months and the patient died three years and eight months after operation. Gastric endocrine cell carcinoma is known as a potentially highly malignant tumor. But in our case FU+CPT-11 controlled growth of the recurrent tumor. Based on this finding, we recommend adjuvant chemotherapy by FU+CPT-11 for gastric endocrine cell carcinoma. PMID- 20009472 TI - [Successful resection of an advanced gastrointestinal stromal tumor by neoadjuvant chemotherapy with imatinib- a case report]. AB - We report a case of a large gastric gastrointestinal stromal tumor (GIST), which we were able to curatively resect after treatment with a daily dosage of 400 mg imatinib for 3 months. The patient was a 46-year-old man whose chief complaint was anemia. Historical diagnosis by endoscopic biopsy was a c-kit-positive GIST of the stomach. From a CT scan, it was suspected that the tumor had directly invaded the pancreas. The tumor was 9 cm in size. For this case, total gastrectomy with distal pancreato splenectomy was necessary for curative resection. Imatinib mesilate was administered as neoadjuvant therapy according to the NCCN guidelines. After 3 months of treatment, CT revealed a dramatic reduction in tumor diameter of 61% and showed direct invasion of the pancreas. The radical operation was considered feasible and a partial gastrectomy was performed. The tumor did not invade other organs, and radical surgery was possible without rupture. The patient was recurrence free at his 12-month follow up examination. PMID- 20009473 TI - [Carcinomatous meningitis in gastric cancer under chemotherapy-two cases]. AB - The development of chemotherapy to treat gastric cancer has prolonged its prognosis, and sometimes extremely rare conditions arise. This report describes two patients with carcinomatous meningitis who rapidly progressed into unconsciousness and died. A 60-year-old woman under second-line chemotherapy for gastric cancer presented with headache and disordered speech. Magnetic resonance imaging (MRI) indicated carcinomatous meningitis, and a lumbar puncture revealed cancer cells in the cerebrospinal fluid. Convulsions rapidly progressed while unconscious, and the patient died two weeks after admission. A 67-year-old man receiving adjuvant chemotherapy after total gastrectomy for gastric cancer was admitted to the emergency department with severe fatigue and appetite loss. Unconsciousness rapidly progressed on the following day and carcinomatous meningitis was diagnosed. The patient died two weeks later. Carcinomatous meningitis is a rare complication of gastric cancer that rapidly progresses often to death, and it should be recalled that it is accompanied by a rapid loss of consciousness. PMID- 20009474 TI - [A case of advanced gastric cancer with disseminated carcinomatosis of bone marrow treated by S-1 and CDDP]. AB - A 33-year-old man was admitted to our hospital due to DIC and multiple bone metastasis after distal gastrectomy for gastric cancer (Stage IIIB). We diagnosed disseminated carcinomatosis of bone marrow by gastric cancer. The patient was treated with combination chemotherapy of S-1 and CDDP (S-1 80 mg/m (2), po, day 1 21 and CDDP 60 mg/m(2), iv, day 8). After one course of the treatment, DIC was resolved and severe pain in his back and legs which had been poorly controlled was dramatically improved. He could thus be discharged from our hospital and survived for about six months. S-1 and CDDP therapy are considered to be effective for disseminated carcinomatosis of bone marrow due to gastric cancer, even if complicated by DIC. PMID- 20009475 TI - [First reported case of lung metastasis from hepatocellular carcinoma successfully treated by ultrasound-guided radiofrequency ablation using ultrasonic contrast agent]. AB - BACKGROUND: Radiofrequency ablation (RFA) is becoming one of the useful options as a local control therapy for lung cancer. Almost all reported cases according to the RFA for lung cancer were performed by CT -guided technique. Only a limited number of articles have been published on ultrasound-guided RFA for lung cancer. CASE: An 80-year-old man underwent transcatheter arterial embolization (TAE) and RFA for hepatocellular carcinoma (HCC) in 2004. A 3.3 cm pulmonary nodule was pointed out in his right lower lobe on a chest CT examination in 2007. The nodule was diagnosed as a lung metastasis from HCC by core needle biopsy. He underwent CT-guided RFA. After three months, the lung metastasis progressed in the same location on the enhanced CT. Then an ultrasound-guided RFA using ultrasonic contrast agent (Sonazoid) was performed. We could distinguish between the necrotic lesion and the viable lesion using Sonazoid, and selective RFA for viable lesion became possible. CONCLUSION: Sonazoid has been highly evaluated in the ultrasound-guided RFA for HCC. To the best of our knowledge, this is the first report of a lung cancer case successfully treated with ultrasound-guided RFA using Sonazoid. PMID- 20009476 TI - [A case of cancer successfully treated by chemotherapy of S-1 as first-line chemotherapy]. AB - The patient was an 83-year-old man who had undergone pancreaduodenectomy for a diagnosed lower bile duct cancer in April 2004. CT demonstrated intra-abdominal lymph node enlargement 2 years postoperatively and CA19-9 increased. S-1, an oral fluoropyrimidine anticancer drug, was started as first-line chemotherapy. A complete response was confirmed after the first two courses of S-1 monotherapy. CR persisted for 2 years and S-1 was discontinued after 25th course because of long CR. No grade 3 or more toxicities were noted. PMID- 20009477 TI - [A case of interstitial pneumonitis induced by FOLFIRI+bevacizumab combination therapy for liver and lung metastasis of colon cancer]. AB - A 64 -year-old female received oral S-1 chemotherapy followed by mFOLFOX6 chemotherapy for postoperative liver and lung metastasis of sigmoid colon cancer. The tumor progression was observed after twelve courses of mFOLFOX6 chemotherapy, and then FOLFIRI+bevacizumab chemotherapy was performed. After two courses of FOLFIRI+bevacizumab chemotherapy, leucopenia was observed. The chemotherapy was then discontinued and G-CSF was administered. Two days later she complained of high fever and dry cough, and was admitted to the hospital. A diffuse ground glass appearance of bilateral lung was observed on chest X-ray and CT. Drug induced interstitial pneumonitis was suspected, and Pneumocystis carini pneumonia was considered in the differential diagnosis. Oral administration of prednisolone and sulfamethoxazole/trimethoprim did not improve the symptoms, so steroid pulse therapy was performed. Steroid pulse therapy improved respiratory symptoms, but CT findings did not change remarkably. After nine weeks in the hospital, she was discharged with home oxygen therapy. Interstitial pneumonitis induced by FOLFIRI+bevacizumab chemotherapy is rare, but the number of cases may increase with increased use of this regimen. The possibility of interstitial pneumonitis should always be considered when the patient presents with a respiratory disorder while receiving systemic chemotherapy. PMID- 20009478 TI - [Effect on neoadjuvant chemotherapy using s-1 for unresectable liver metastasis from rectal cancer-a case report]. AB - The patient was a 42-year-old female who had advanced rectal cancer(Ra, type 2)with liver metastasis. The preoperative diagnosis was T2N1H2, but the liver tumor was unresectable for hepatic vein invasion in September, 2004. She underwent low anterior resection for primary rectal cancer at first after she was given S-1 80 mg/day(14 days)for a histological effect judgment. The pathological diagnosis was moderately-differentiated adenocarcinoma, ss, n1, and the histological effect was Grade 1a. Because CT showed reduction of liver metastasis after the operation, she was treated with 2 courses of chemotherapy using S-1. Radiographic examination showed reduction of liver metastasis(36%)and improvement of hepatic vein invasion. In January, 2005, she underwent hepatectomy and the pathological diagnosis was moderately-differentiated adenocarcinoma, metastasis and the histological effect was Grade 1a. There has not been any recurrence for 48 months after hepatectomy. We experienced this case in which preoperative S-1 proved effective for liver metastasis from rectal cancer. PMID- 20009479 TI - Issues in the design of molecular and genetic epidemiologic studies. AB - The final decision of study design in molecular and genetic epidemiology is usually a compromise between the research study aims and a number of logistical and ethical barriers that may limit the feasibility of the study or the interpretation of results. Although biomarker measurements may improve exposure or disease assessments, it is necessary to address the possibility that biomarker measurement inserts additional sources of misclassification and confounding that may lead to inconsistencies across the research literature. Studies targeting multi-causal diseases and investigating gene-environment interactions must not only meet the needs of a traditional epidemiologic study but also the needs of the biomarker investigation. This paper is intended to highlight the major issues that need to be considered when developing an epidemiologic study utilizing biomarkers. These issues covers from molecular and genetic epidemiology (MGE) study designs including cross-sectional, cohort, case-control, clinical trials, nested case-control, and case-only studies to matching the study design to the MGE research goals. This review summarizes logistical barriers and the most common epidemiological study designs most relevant to MGE and describes the strengths and limitations of each approach in the context of common MGE research aims to meet specific MEG objectives. PMID- 20009480 TI - Identification and application of biomarkers in molecular and genomic epidemiologic research. AB - Biomarkers are characteristic biological properties that can be detected and measured in a variety of biological matrices in the human body, including the blood and tissue, to give an indication of whether there is a threat of disease, if a disease already exists, or how such a disease may develop in an individual case. Along the continuum from exposure to clinical disease and progression, exposure, internal dose, biologically effective dose, early biological effect, altered structure and/or function, clinical disease, and disease progression can potentially be observed and quantified using biomarkers. While the traditional discovery of biomarkers has been a slow process, the advent of molecular and genomic medicine has resulted in explosive growth in the discovery of new biomarkers. In this review, issues in evaluating biomarkers will be discussed and the biomarkers of environmental exposure, early biologic effect, and susceptibility identified and validated in epidemiological studies will be summarized. The spectrum of genomic approaches currently used to identify and apply biomarkers and strategies to validate genomic biomarkers will also be discussed. PMID- 20009481 TI - Discovering gene-environment interactions in the post-genomic era. AB - In the more than 100 genome wide association studies (GWAS) conducted in the past 5 years, more than 250 genetic loci contributing to more than 40 common diseases and traits have been identified. Whilst many genes have been linked to a trait, both their individual and combined effects are small and unable to explain earlier estimates of heritability. Given the rapid changes in disease incidence that cannot be accounted for by changes in diagnostic practises, there is need to have well characterized exposure information in addition to genomic data for the study of gene-environment interactions. The case-control and cohort study designs are most suited for studying associations between risk factors and occurrence of an outcome. However, the case control study design is subject to several biases and hence the preferred choice of the prospective cohort study design in investigating gene-environment interactions. A major limitation of utilising the prospective cohort study design is the long duration of follow-up of participants to accumulate adequate outcome data. The GWAS paradigm is a timely reminder for traditional epidemiologists who often perform one- or few-at-a-time hypothesis testing studies with the main hallmarks of GWAS being the agnostic approach and the massive dataset derived through large-scale international collaborations. PMID- 20009482 TI - Gene-diet interaction on cancer risk in epidemiological studies. AB - Genetic factors clearly play a role in carcinogenesis, but migrant studies provide unequivocal evidence that environmental factors are critical in defining cancer risk. Therefore, one may expect that the lower availability of substrate for biochemical reactions leads to more genetic changes in enzyme function; for example, most studies have indicated the variant MTHFR genotype 677TT is related to biomarkers, such as homocysteine concentrations or global DNA methylation particularly in a low folate diet. The modification of a phenotype related to a genotype, particularly by dietary habits, could support the notion that some of inconsistencies in findings from molecular epidemiologic studies could be due to differences in the populations studied and unaccounted underlying characteristics mediating the relationship between genetic polymorphisms and the actual phenotypes. Given the evidence that diet can modify cancer risk, gene-diet interactions in cancer etiology would be anticipated. However, much of the evidence in this area comes from observational epidemiology, which limits the causal inference. Thus, the investigation of these interactions is essential to gain a full understanding of the impact of genetic variation on health outcomes. This report reviews current approaches to gene-diet interactions in epidemiological studies. Characteristics of gene and dietary factors are divided into four categories: one carbon metabolism-related gene polymorphisms and dietary factors including folate, vitamin B group and methionines; oxidative stress-related gene polymorphisms and antioxidant nutrients including vegetable and fruit intake; carcinogen-metabolizing gene polymorphisms and meat intake including heterocyclic amins and polycyclic aromatic hydrocarbon; and other gene diet interactive effect on cancer. PMID- 20009483 TI - Risk assessment and pharmacogenetics in molecular and genomic epidemiology. AB - In this article, we reviewed the literature on risk assessment (RA) models with and without molecular genomic markers and the current utility of the markers in the pharmacogenetic field. Epidemiological risk assessment is applied using statistical models and equations established from current scientific knowledge of risk and disease. Several papers have reported that traditional RA tools have significant limitations in decision-making in management strategies for individuals as predictions of diseases and disease progression are inaccurate. Recently, the model added information on the genetic susceptibility factors that are expected to be most responsible for differences in individual risk. On the continuum of health care, from diagnosis to treatment, pharmacogenetics has been developed based on the accumulated knowledge of human genomic variation involving drug distribution and metabolism and the target of action, which has the potential to facilitate personalized medicine that can avoid therapeutic failure and serious side effects. There are many challenges for the applicability of genomic information in a clinical setting. Current uses of genetic markers for managing drug therapy and issues in the development of a valid biomarker in pharmacogenetics are discussed. PMID- 20009484 TI - Assessment of village health worker training program in Tuguegarao, Philippine. AB - OBJECTIVES: This study was performed to evaluate the effectiveness of 'village health worker training program' which aimed to build community participatory health promotion capacity of community leaders in villages of low developed country and to develop methods for further development of the program. METHODS: The intervention group were 134 community leaders from 25 barangays (village). Control group were 149 form 4 barangays. Intervention group participated 3-day training program. Questionnaire was developed based on 'Health Promotion Capacity Checklist' which assessed capacity in 4 feathers; 'knowledge', 'skill', 'commitment', and 'resource'. Each feather was assessed in 4 point rating scale. Capacity scores between intervention group and control group were examined to identify changes between the pre- and post-intervention periods. A qualitative evaluation of the program was conducted to assess the appropriateness of the program. The program was conducted in Tuguegarao city, Philippine in January, 2009. RESULTS: The result showed significant increases in the total health promotion capacity and each feather of health promotion capacities between pre and post assessment of intervention group. But there was no significant change in that of control group. Participants marked high level of satisfaction for preparedness, selection of main subjects and education method. Qualitative evaluation revealed that training program facilitated community participatory health promotion capacity of participants. CONCLUSIONS: This study suggested that the Village health worker training program is effective for building health promotion capacity of community leaders and it can be a main method for helping low developed countries with further development. PMID- 20009485 TI - [The association of central obesity with type 2 diabetes among Koreans according to the serum gamma-glutamyltransferase level: Korean genome and epidemiology study]. AB - OBJECTIVES: This cross-sectional study was performed to examine if the serum gamma-glutamyltransferase (GGT) level that is within its normal range is associated with the risk of type 2 diabetes and if the association between the waist hip ratio (WHR) and type 2 diabetes is different depending on the serum GGT levels. METHODS: The study subjects were 23,436 persons aged 40 years or older and who participated in regular health check-ups at 11 hospitals (males: 5,821, females: 17,615). The gender-specific quintiles of the serum GGT and WHR were used to examine the associations with type 2 diabetes. RESULTS: The serum GGT levels within their normal range were positively associated with type 2 diabetes only in women. The adjusted odds ratios (ORs) were 1.0, 1.0, 1.4, 2.1, and 2.5 according to the quintiles of the serum GGT (p(trend)<0.01). The WHR was more strongly associated with the prevalence of diabetes among the women with a high normal serum GGT level as compared with those with a low-normal serum GGT level (p for interaction=0.02). For example, the adjusted ORs for women with a low normal serum GGT level were 1.0, 1.2, 1.5, 2.2, and 2.4 according to the quintiles of the WHR, while those figures were 1.0, 2.4, 3.6, 5.0, and 8.3 among the women with a high normal serum GGT level. However, in men, the serum GGT was very weakly associated with type 2 diabetes and the association between the WHR and type 2 diabetes was not different depending on the serum GGT level. CONCLUSIONS: Serum GGT within its normal range was positively associated with type 2 diabetes, and central obesity was more strongly associated with the prevalence of type 2 diabetes when the serum GGT level was high-normal. However, these associations were observed only in women, which is different from the previous findings. The stronger relation between central obesity and type 2 diabetes among women with a high-normal serum GGT level can be useful for selecting a group that is at high risk for type 2 diabetes irregardless of whatever the underlying mechanism is. PMID- 20009486 TI - [Development of composite deprivation index for Korea: the correlation with standardized mortality ratio]. AB - OBJECTIVES: The aims of this paper were to develop the composite deprivation index (CDI) for the sub-district (Eup-Myen-Dong) levels based on the theory of social exclusion and to explore the relationship between the CDI and the standardized mortality ratio (SMR). METHODS: The paper calculated the age adjusted SMR and we included five dimensions of social exclusion for CDI; unemployment, poverty, housing, labor and social network. The proxy variables of the five dimensions were the proportion of unemployed males, the percent of recipients receiving National Basic Livelihood Security Act benefits, the proportion of households under the minimum housing standard, the proportion of people with a low social class and the proportion of single-parent household. All the variables were standardized using geometric transformation and then we summed up them for a single index. The paper utilized the 2004-2006 National Death Registry data, the 2003-2006 national residents' registration data, the 2005 Population Census data and the 2005-2006 means-tested benefit recipients' data. RESULTS: The figures were 115.6, 105.8 and 105.1 for the CDI of metropolitan areas (big cities), middle size cities and rural areas, respectively. The distributional variation of the CDI was the highest in metropolitan areas (8.9 - 353.7) and the lowest was in the rural areas (26.8 - 209.7). The extent and relative differences of deprivation increased with urbanization. Compared to the Townsend and Carstairs index, the CDI better represented the characteristics of rural deprivation. The correlation with the SMR was statistically significant and the direction of the CDI effects on the SMR was in accordance with that of the previous studies. CONCLUSIONS: The study findings indicated mortality inequalities due to the difference in the CDI. Despite the attempt to improve deprivation measures, further research is warranted for the consensus development of a deprivation index. PMID- 20009487 TI - [The contributing factors to surplus medicine by long-term users of medical aid in Korea]. AB - OBJECTIVES: The amount of medical utilization by Medical Aid recipients was 3.7 times that of patients with Korean Medical Insurance. This study aims to describe the surplus medicine and the medication-related utilization, and to determine factors contributing to surplus medicine. METHODS: Among those who used copayment free Class I Medical Aid in 2005, 146,880 subjects who were >/=19 year-old and received >365 days medical treatment per year were studied with their case managers by conducting face-to-face interviews. The analytic methods were description, chi-square, t-tests, ANCOVA and multiple logistic regressions. RESULTS: Most subjects were female (68.6%), the elderly (62.5%), and the separated (61.6%), had an elementary graduation or less (74.8%), and had disabilities (33.2%). The percentage of subjects with surplus medicine was 18.5%. However, the percentage of females, the elderly, those with non-disabilities, the separated, the uneducated, those with a very poor perceived health status and those with an economical burden for medical treatment was 19.3%, 18.9%, 19.0%, 19.3%, 19.0%, 20.2% and 24.3%, respectively. For subjects with surplus medicine, averages for the number of used pharmacies, the pharmacy-visit days and the medication costs were 4.6 drugstores, 34.9 days and approximately 1,124 thousand Won(?). These values were higher than those without surplus medicine (4.4 drugstores , 33.8 days, and Won(?)1,110 thousand, respectively). The odds ratios of the contributing factors to surplus medicine were female 1.11 (95% CI=1.07 1.14), the elderly 1.06 (95% CI=1.02-1.10), those with non-disabilities 1.08 (95% CI=1.05-1.12), the separated 1.14 (95% CI=1.10-1.18), the unmarried 1.12 (95% CI=1.07-1.18), the uneducated 1.03 (95% CI=1.01-1.08), those with a very poor perceived health status 1.04 (95% CI=1.01-1.08) and experiencing an economical burden for medical treatment 2.33 (95% CI=2.26-2.40). CONCLUSIONS: 18.5% of subjects had surplus medicine with a higher mean of medication cost. Therefore, health education and health promotion programs to prevent surplus medicine and to improve the appropriate usage of medication are necessary. PMID- 20009488 TI - [Mumps transmission control status and inapparent infection rate among middle and high school students during the 2007-2008 mumps outbreak in Daegu]. AB - OBJECTIVES: This study was performed to investigate the mumps transmission control status and inapparent infection rate among middle and high school students in Daegu City during a mumps outbreak. METHODS: Nine schools (two middle schools and seven high schools), which reported a number of mumps cases between 2007 and 2008 were selected for investigation. During March-May 2008, a standard questionnaire was distributed to gather information about case identification, instructed isolation measure, isolation status of mumps cases and related factors, and outdoor activities of non-isolated mumps case. Inapparent infection rate was estimated by serum mumps IgM and IgG antibodies status and self-reported mumps symptoms in three of the nine schools. RESULTS: Among 2,560 respondents, more than half of students answered that they did not receive instructions in mumps transmission control measures during the outbreak. Among the 327 mumps cases identified by the questionnaire, 131 cases (40.1%) were considered as isolated and the isolation rates were significantly different among schools, grades, and gender. Of the non-isolated cases, 88.3% continued attending school. Inapparent mumps infection rates were between 56.3% and 70.2%. CONCLUSIONS: Mumps transmission control was inadequate to control the mumps outbreak. Although high inapparent infection rate would mitigate the transmission control effect of case isolation, this measure is fundamental for infection control. The reasons of this inadequate status need to be explored to develop an effective intervention strategy. PMID- 20009489 TI - [Medical care utilization status and associated factors with extended hospitalization of psychiatric patients in Korea]. AB - OBJECTIVES: This study was performed to examine medical care utilization of psychiatric patients and to explore patients' characteristics associated with extended hospitalization. METHODS: Data were extracted from information of Korean Health Insurance Review and Assessment Service. All data associated with admission and outpatient clinic visit were analysed by patient characteristics. We selected first psychiatric admission patients who diagnosed mental and behavioral disorders due to use of alcohol (main disease code: F10), schizophrenia and related disorders (F20-29) and mood disorders (F30~33) from January to June 2005. We analysed status of admission, mean length of stay, regular access to outpatient clinic and rates of extended hospitalization during 3 years. Bivariate and multivariate analyses were conducted to identify factors associated with extended hospitalization. RESULTS: The number of psychiatric patients during the first six month of 2005 was 30,678. The mean length of stay was longest for schizophrenia and related disorders but shortest for mood disorders. Patients who experienced an extended hospitalization were 18.8% of total subjects. An extended hospitalization was more common in schizophrenia and related disorders than other diagnostic groups. The factors associated with the extended hospitalization were age, sex, diagnostic group, type of insurance and medical care utilization groups. CONCLUSIONS: The study indicates the problem of an extended hospitalization for psychiatric patients in Korea. It is suggested that variations in rates of extended hospitalization among medical care utilization group may need an active early intervention system in psychiatric treatment service. Particular attention needs to be devoted to planning and funding for reducing extended hospitalization. PMID- 20009490 TI - Endocrinology and art. Bronzino or Pontormo? Look at the neck! PMID- 20009491 TI - Does prolactin induce apoptosis? Evidences in a prostate cancer in vitro model. AB - BACKGROUND: Prolactin (PRL) regulates prostate growth and differentiation. Some studies have suggested that PRL has a pro-apoptotic effect on a myeloma cell line and in newt spermatogonia. The proliferative effect of PRL on prostate cancer cell lines is, however, a controversial area. AIM: On this account, we evaluated the effects of PRL on the prostate cancer cell lines LNCaP and PC3 apoptosis and proliferation. MATERIALS AND METHODS: LNCaP and PC3 cells were exposed to increasing concentrations of PRL for 24, 48, 72 and 96 hours. Staining with propidium iodide (PI) and TUNEL assay followed by flow cytometry were used to detect apoptosis. LNCaP and PC3 proliferation was assessed by optical microscopy counting. RESULTS: PRL induced a dose-dependent decrease of DNA content and an increase of DNA fragmentation in LNCaP after 96 hours of incubation. These effects were observed with physiological concentrations of PRL and were counteracted by a prolactin receptor antagonist. On the other hand, PRL did not have any effect on DNA content or fragmentation in PC3 cells. No effect of PRL on LNCaP and PC3 proliferation was found. CONCLUSIONS: This study indicates that PRL induces apoptosis in the androgen-responsive cell line LNCaP, whereas no effect was observed in the androgen-insensitive PC3 cell line. These findings suggest that androgen responsiveness may be required for PRL to be effective on prostatic cells. PMID- 20009492 TI - Erythrocyte fatty acid composition and insulin sensitivity in daughters of Type 2 diabetic patients and women with no family history of diabetes. AB - BACKGROUND: There is evidence that the impact of environmental factors on insulin sensitivity is modified by the presence of family history of diabetes. AIM: To compare the association between the erythrocyte phospholipid fatty acid composition (a biomarker of dietary fatty acids) and insulin sensitivity in daughters of Type 2 diabetic patients with the corresponding association in women without family history of diabetes. MATERIAL/SUBJECTS AND METHODS: Eighteen offspring of Type 2 diabetic patients [age 30+/-6.5 yr; body mass index (BMI) 22.2+/-2.5 kg/m2; body fat 31.8+/-5.1%] and 18 matched women (age 30.1+/-6.8 yr; BMI 22.2+/-1.8 kg/m2; body fat 32.2+/-6.0%) participated in the study. RESULTS: Insulin Sensitivity Index (ISI)-Matsuda tended to be lower (p=0.06) in the Offspring than the control group. Weight proportions of erythrocyte phospholipid saturated (SFA), polyunsaturated (PUFA), and monounsaturated fatty acids (MUFA) were similar between the two groups. In the offspring, erythrocyte total SFA were negatively correlated with ISI-Matsuda [r=-0.47, p<0.05), ISI(gly)-Belfiore (r= 0.52, p<0.05) and ISI(ffa)-Belfiore (r=-0.53, p<0.05)], whereas total PUFA were positively correlated with insulin sensitivity [ISI-Matsuda, r=0.46, p<0.05; ISI(gly)-Belfiore, r=0.53, p<0.05; ISI(ffa)-Belfiore, r=0.54, p<0.05]. No significant correlations were observed in the control group. CONCLUSIONS: The associations between erythrocyte fatty acid composition and insulin sensitivity are distinct between daughters of Type 2 diabetic patients and women without family history of diabetes. PMID- 20009493 TI - BRAFV600E mutation in the pathogenesis of a large series of papillary thyroid carcinoma in Czech Republic. AB - BACKGROUND: Activating point mutation of the BRAF gene, the most common genetic alteration reported in papillary thyroid carcinomas (PTC), has been associated with poor prognostic characteristics. AIM: Our objective was to determine the frequency of BRAFV600E mutation in PTC tumor tissues from the period 1960-2007 and to correlate it with clinicopathological parameters. SUBJECTS AND METHODS: DNAs were extracted from 242 PTCs, 23 sporadic medullary carcinomas, one anaplastic carcinoma and 6 poorly differentiated carcinomas. The presence of BRAFV600E mutation was determined using single strand conformation polymorphism method and verified by direct sequencing. RESULTS: BRAFV600E mutation was detected in 81 of 242 PTCs (33.5%), in one of 6 poorly differentiated carcinomas (16.7%) and in anaplastic carcinoma. BRAFV600E mutation was much less frequent in the follicular variant compared to classical variant and mixed follicular- classical variant of PTCs (p=0.001). BRAFV600E mutation was significantly associated with presence of nodal metastasis (p=0.029), more advanced TNM stage (p=0.014) and recurrence of disease (p=0.008). The mutation correlated with a higher age at diagnosis (p=0.049) and with a greater tumor size (p=0.041). Multivariate analysis confirmed these findings. The prevalence of BRAFV600E mutation before 1986 was significantly lower than after it (p=0.008). CONCLUSIONS: Our data suggest that BRAFV600E mutation is associated with high risk clinicopathological characteristics of PTC and worse prognosis of patients. The frequency of the mutation significantly varied during the observed period but rather because of the different age distribution of patients in particular periods than as a consequence of Chernobyl accident. PMID- 20009495 TI - Comparison of epicardial and myocardial perfusions after primary coronary angioplasty for ST-elevation myocardial infarction in patients under and over 75 years of age. AB - BACKGROUND AND AIMS: Patients aged >=75 years compose a high-risk subgroup for acute myocardial infarction (AMI). It is unknown whether myocardial perfusion in these patients is decreased compared with younger ones after primary percutaneous coronary intervention (PPCI), which may contribute to their worse prognosis. We compared epicardial and myocardial perfusions as well as short-term outcomes between elderly and younger patients undergoing PPCI. METHODS: A total of 547 consecutive PPCI patients were prospectively enrolled; of these, 106 were elderly (>=75 yrs). Epicardial perfusion was evaluated by the Thrombolysis in Myocardial Infarction (TIMI) flow grade and corrected TIMI frame count (CTFC), and myocardial perfusion was evaluated by the TIMI myocardial perfusion grade (TMPG) and ST-segment resolution (STR). RESULTS: Despite comparable epicardial perfusion pre- and post-PPCI, elderly patients had impaired myocardial perfusion after PPCI, as measured by reduced TMPG (35.9% vs 14.5%, p=0.001) and absent STR (18.9% vs 9.8%, p=0.009). After adjusting for clinical and angiographic risk profiles, multivariate analysis showed that age >=75 years remained independently associated with reduced TMPG or absent STR. In the whole population, multivariate analysis revealed that both age >=75 years and absent STR were independently associated with 3-month major adverse cardiac events (MACE). In the elderly subgroup, multivariate analysis identified absent STR as the strongest determinant of 3-month MACE. CONCLUSIONS: Age is associated with impaired myocardial perfusion, but not epicardial perfusion, after PPCI for AMI. To further improve the outcome of elderly AMI patients, efforts should be aimed at improving myocardial perfusion beyond epicardial recanalization. PMID- 20009494 TI - Comparison of pegvisomant and long-acting octreotide in patients with acromegaly naive to radiation and medical therapy. AB - BACKGROUND: Normalization of IGF-I in patients with acromegaly is associated with a decrease in mortality. Pegvisomant may be more effective in lowering IGF-I than octreotide. SUBJECTS AND METHODS: The efficacy and safety of pegvisomant and octreotide long-acting release (LAR) were compared in 118 patients with acromegaly in this 52-week, multicenter, open-label, randomized study. The primary endpoint was IGF-I normalization at week 52. Secondary endpoints included mean changes from baseline in IGF-I, IGF binding protein 3, acromegaly signs and symptom scores, ring size, acromegaly quality of life questionnaire scores, and safety. RESULTS: Fifty-six patients received pegvisomant and 57 received octreotide LAR. IGF-I normalized in 51% of pegvisomant patients and 34% treated with octreotide LAR (p=0.09, ns). Patients with baseline IGF-I > or = 2x upper limit of normal had a higher rate of IGF-I normalization with pegvisomant vs octreotide LAR (p=0.05). Among the patients who did not achieve a normalized IGF I, pegvisomant-treated patients were more likely to be receiving < 30 mg of study drug (71% vs 16%). Treatment-related adverse events were mild-to-moderate in both groups. Mean fasting glucose decreased in diabetic and non-diabetic patients on pegvisomant whereas octreotide LAR was associated with an increase at week 52 (p=0.005 and p=0.003 between groups, respectively). Mean change in tumor volume during treatment was similar between groups. CONCLUSIONS: Pegvisomant and octreotide LAR were equally effective in normalizing IGF-I in the overall population, and pegvisomant was more effective in patients with higher baseline IGF-I levels. Pegvisomant had a more favorable effect on parameters of glycemic control. PMID- 20009496 TI - Quality of life three years after major lower extremity amputation due to peripheral arterial disease. AB - BACKGROUND AND AIMS: The purpose of this study was to assess the quality of life (QoL) of peripheral arterial disease (PAD) amputees. PATIENTS AND METHODS: Fifty nine PAD patients (mean age 75.2, SD 10.7, range 39-96, 47% men) who had undergone their first major lower leg amputation (LEA) on average 2.7 years earlier (in 1998-2002) were interviewed, and 118 age- and gender-matched controls completed a postal questionnaire. Outcomes were assessed with 15D Health-Related QoL instrument, Rand-36 Physical Functioning- and General Health subscales, Geriatric Depression Scale, 6-item Brief Social Support Questionnaire, and Self reported Life Satisfaction score. RESULTS: The amputees had more diseases than their controls. HRQoL was lower among amputees than among controls. Half the amputees lived in institutional care, 25% had a Mini-Mental Examination score <18, and 22% had unilateral belowknee amputations only. The amputees had a similar self-assessed sense of their general state of health, life satisfaction and perceived social support as controls. Amputees who were institutionalized and those who did not use prostheses had more symptoms of depression than those who lived at home or used prostheses. CONCLUSION: Home-dwelling amputees had a relatively good QoL, whereas institutionalization was associated with depressive symptoms. In rehabilitation programs, not only physical disability assessment but also QoL should be considered. PMID- 20009497 TI - Effects of a dynamic balance training protocol on podalic support in older women. Pilot Study. AB - BACKGROUND AND AIMS: The foot provides the only direct contact with supporting surfaces and therefore plays an important role in all postural tasks. Changes in the musculoskeletal and neurological characteristics of the foot with advancing age can alter plantar loading patterns and postural balance. Several studies have reported that exercise training improves postural performance in elderly individuals. The aim of our study was to investigate the effectiveness of a dynamic balance training protocol performed for 5 weeks on the support surface, percentage distribution of load in both feet, and body balance performance in healthy elderly women. METHODS: Ten subjects (68.67+/-5.50 yrs old; 28.17+/-3.35 BMI) were evaluated with a monopodalic performance test and baropodometric analyses before and after the training period. RESULTS: We found a significant improvement in balance unipedal performance times on left and right foot by 20.18% and 26.23% respectively (p<0.05). The support surface of the right foot significantly increased in response to the training protocol and, in particular, in both forefoot and rearfoot regions (p<0.05). In addition, before the training period, load distribution on the left foot was greater than on the right one; equal load redistribution was measured on both feet in response to exercise (p>0.05). CONCLUSIONS: The increased support surface and equal redistribution of body weight on both feet obtained in response to our training protocol may be postural adaptations sufficient to improve static balance in elderly women. PMID- 20009498 TI - What is the role played by physical activity and exercise in the frailty syndrome? Perspectives for future research. AB - Exercise and physical activity play an important role in physical frailty, but we do not know if they are markers, components and/or correlates of this syndrome. The purpose of this paper is briefly to discuss the potential roles played by physical activity and exercise on the development and progression of frailty, and to propose directions for future research in this field. Exercise practice lowers the levels of some frailty markers, such as tumor necrosis factor-alpha, interleukin-6, C-reactive protein, and uric acid, and also resistance to insulin. The influence of exercise on the main frailty domains is also well established in the literature. Exercise improves muscle strength, gait speed, cognition (particularly executive control related-tasks), weight maintenance, mood and, to a lesser extent, feelings of energy. Although exercise and physical activity positively influence the main frailty markers and domains, most findings were obtained for other elderly populations (e.g., healthy elderly, clinical populations). For future research, efforts must be made to define some key concepts (exercise or physical activity) in selecting study samples and in establishing intervention length. Attention must also be paid to identifying the most efficacious exercise interventions regarding type, frequency, intensity and session duration, and approaching a dose-response relationship between a physically active life-style and frailty. Thus, further research, especially longitudinal randomized controlled trials, is needed to understand the role of physical activity and exercise in the frailty syndrome. PMID- 20009499 TI - Games-based biofeedback training and the attentional demands of balance in older adults. AB - BACKGROUND AND AIMS: Programs designed to improve balance in older adults may function by improving general fitness (strength, endurance, range of motion) and also changing the attentional demands of postural control. Research in previously sedentary older adults cannot differentiate between changes in balance ability resulting from improved fitness or reduced attentional demands. A training program of games-based balance biofeedback was given to nine older adults with previous exercise experience. METHODS: Training consisted of sixteen sessions (twice weekly for eight weeks) of 30 minutes each. Postural sway (force plate measurement), attentional demands (dual task paradigm), the Community Balance and Mobility Scale (CB&M), and the six minute walk test were measured in pre, post and retention tests. RESULTS: Participants in the training group significantly decreased their reaction time from pre to post testing in a dual task paradigm compared to a control group. The training group also significantly increased their scores on the CB&M scale compared to control participants. The decreased reaction times and increased CB&M scores observed in the training group were maintained through a two week retention period. Changes in reaction time were significantly correlated with changes in CB&M score. Six minute walk distance increased significantly in both groups and did not appear to result directly from the training program. CONCLUSIONS: Gamesbased balance biofeedback training using a range of training postures can significantly improve functional balance in exercise trained older adults by reducing the attentional demands of postural control. PMID- 20009500 TI - Plasma nitrite response in older women to a physical function test. AB - BACKGROUND AND AIMS: Nitric oxide (NO) may play a critical role in facilitating the delivery of blood to active skeletal muscle, ultimately impacting functional health in older adults. Plasma nitrite is a useful marker of vascular NO bioavailability. The aim of the current investigation was to examine the effect of a widely used physical function test on plasma nitrite concentrations in older adults. METHODS: Venous blood was drawn before, immediately following, and 10 minutes following the completion of a 400-m walk test. Blood samples were added to heparin and frozen for subsequent analysis of nitrite levels using chemiluminescence. RESULTS: Twenty six (79+/-4 yrs) women participated in this study. Plasma nitrite levels decreased approximately 22% from baseline following a 400-m walk. Percent change in plasma nitrite was related to walking speed (r= 0.550, p=0.004). CONCLUSIONS: These data suggest an alteration in plasma nitrite concentration following a functional test which may impact functional health. PMID- 20009502 TI - Biphasic effects of dexamethasone on glycogen metabolism in primary cultured rat hepatocytes. AB - Glucocorticoids (GC), the basic function of which is modulating carbohydrates metabolism, play a critical role in stress response by enhancing the organism's resistance. It is widely believed that they could promote glycogen synthesis. However, it is doubtful whether GC can still stimulate glycogen deposition in stress response, as it is known that glucose is imperatively needed at that time. Here, we used primary cultured rat hepatocytes to investigate the effects of GC on glycogen metabolism in vitro to exclude other influences in stress. The results showed that dexamethasone (Dex) played biphasic effects on hepatocytes glycogen metabolism depending on its dosage and the duration of stimulation. Dex could decrease glycogen content of hepatocytes in the higher concentration within a relatively shorter period of time, which could not be blocked by cycloheximide. Therefore, dual roles in hepatic glycogen metabolism played by GC were demonstrated, and a non-genomic mechanism might be involved in the glycogenolytic action of GC. We postulated that the biphasic effects of GC on hepatic glycogen metabolism might be of important significance in stress response. PMID- 20009501 TI - Prevalence and risk factors of albuminuria in Type 2 diabetes in Bahrain. AB - Microalbuminuria is the earliest sign of diabetic nephropathy, and is a marker of increased diabetic cardiovascular morbidity and mortality. We investigated the prevalence of albuminuria and associated risk factors among Type 2 diabetes (T2DM) patients, who were attending diabetes clinics at primary healthcare facilities in Bahrain. This was a cross-sectional study on 712 adult T2DM Bahraini patients screened for albuminuria using albumin:creatinine ratio test (ACR), in 9 selected primary healthcare centers in Bahrain in the period February March 2006. Of the 702 participants, 196 (27.9%) were albuminuric. The prevalence of microalbuminuria (ACR>3.4 mg albumin/mmol creatinine) and macroalbuminuria (protein:creatinine ratio >22 mg protein/ mmol creatinine) were 22.0% (no.=155) and 5.8% (no.=41), respectively. Univariate analysis demonstrated significant associations between albuminuria and older age (>65 yr), longer duration of diabetes, high glycated hemoglobin (HbA1c), elevated triglycerides, total- and LDL-cholesterol, systolic (>160 mmHg) and diastolic (>90 mmHg) blood pressure (BP), and retinopathy. Multivariate regression analysis showed that high HbA1c, higher systolic BP, and long duration of diabetes, were significant predictors of albuminuria, while older age, male gender, and higher triglycerides were borderline predictors. Albuminuria in Bahraini T2DM patients was related to poor glycemic control, higher systolic BP, and longer duration of diabetes. Effective preventive and control strategies of those factors should be urgently implemented. PMID- 20009503 TI - Endocrinology and Art. 4 movie actors with acromegaly/gigantism. PMID- 20009504 TI - Translational control of bacterial heat shock and virulence genes by temperature sensing mRNAs. AB - Riboswitches and RNA thermometers are built-in sensory elements that control the fate of mRNAs in response to environmental conditions. Both are comprised of complex RNA structures that undergo a conformational change when a certain chemical or physical signal is present. In bacteria, these elements are usually located in the 5'-untranslated region (5'-UTR). Typical RNA thermometers control translation initiation of heat shock or virulence genes by forming a secondary structure that traps the ribosome binding site (RBS). An increase in temperature to 37 degrees C (virulence genes) or higher (heat shock genes) destabilizes the structure, liberates the RBS and permits formation of the translation initiation complex. Recent findings suggest that RNA thermometers are more widely used than initially anticipated. The molecular details of selected thermometers and the potential advantage of using such riboregulators will be discussed. PMID- 20009505 TI - Similarities and differences between phytochrome-mediated growth inhibition of coleoptiles and seminal roots in rice seedlings. PMID- 20009506 TI - Minting a new class of polo-like-kinase inhibitors. PMID- 20009507 TI - Riboswitch-mediated control of gene expression in eukaryotes. AB - The discovery of metabolite-sensing RNA domains with gene regulatory functions, so-called riboswitches, has greatly expanded our view of the structural and functional complexity of RNA. Hitherto, more than 20 distinct riboswitch classes have been identified, which respond to a variety of small molecules and perform sophisticated gene regulatory tasks. Riboswitches are typically positioned in the 5' untranslated region of bacterial mRNAs, where they control gene expression by transcriptional or translational attenuation. However, the recent investigation of additional riboswitch classes has revealed a more complex repertoire of regulatory mechanisms. This also includes splicing control in filamentous fungi, green algae and higher plants by thiamin pyrophosphate-binding riboswitches, the only class of metabolite-sensing RNAs identified in eukaryotes so far. All eukaryotic riboswitches characterized to date modulate, in a first step, splicing, but the downstream processes under control vary fundamentally among different species further highlighting the versatility of this gene regulatory mechanism. PMID- 20009508 TI - Multiple levels of post-transcriptional control of expression of the poly (A) binding protein. AB - Regulation of gene expression at the post-transcriptional level such as control of mRNA translation and stability is of fundamental importance because it allows cells to respond quickly to external signals, and change protein synthesis without new transcriptional activity. As such, control of translation and stability of mRNA play crucial roles in a variety of cellular processes, including regulating normal cellular growth, embryogenesis and neuronal plasticity. Consequently, misregulation of mRNA translation or degradation can be associated with a number of human diseases, such as cancer and diabetes. Studies have shown that the cytoplasmic poly (A)-binding protein (PABP) plays a crucial role in regulating both translation and stability of eukaryotic mRNA. PMID- 20009509 TI - In vitro studies of the transmission barrier. AB - Protein Misfolding Cyclic Amplification (PMCA) has proved to be an efficient method mimicking in vitro some of the fundamental steps involved in prion replication in vivo. Thus, it can be used to efficiently replicate a variety of prion strains/species. The in vitro generated prions possess key prion features, i.e., they are infectious in vivo and maintain their strain specificity. One of the big challenges is its use for studying prion transmission barriers. PMCA has been efficiently used for adapting different prion species through a range of species barriers; however its capacity for overcoming purportedly unbreakable species barriers compels us to adapt it in order to use it as a reliable technique. In addition, this in vitro method might be a crucial tool in evaluating the potential risks of different prion strains (natural or experimentally generated in vitro) to humans and animals. PMID- 20009510 TI - Evolutional imprints on the sequences of BMP2/4/DPP type proteins. AB - Decapentaplegic (DPP) and bone morphogenetic protein (BMP)-2 and -4 type ligands form a branch of the transforming growth factor-beta (TGFbeta) superfamily. They play prominent roles in metazoan developmental processes as diverse as cell proliferation, apoptosis, differentiation and cell-fate determination. Maturation of the BMP2/4/DPP type proteins requires proteolytic cleavage of their proproteins by furin-type proprotein convertases (PCs). Even though cleavage of the prodomain is critical for signaling, much less attention has been paid to the role of proteolytic processing. Our studies suggest that the cleavage sites of BMP2/4/DPP type proteins have been diversified and can be categorized into at least four different types. These findings indicate that the cleavage sites in BMP2/4/DPP prodomains are tolerant to mutations acquired through evolution and have adapted to diversified environments among species. These results provide insights for further studies of evolution and molecular diversity. In this Extra View we discuss the evolutional variation seen in the sequences of BMP2/4/DPP type proteins. PMID- 20009511 TI - The ever expanding role of aquaglyceroporins: confirmation of protein-facilitated boron transport. PMID- 20009512 TI - TDP1 serine 81 promotes interaction with DNA ligase IIIalpha and facilitates cell survival following DNA damage. AB - Tyrosyl DNA phosphodiesterase (TDP1) is a DNA 3'-end processing enzyme that preferentially hydrolyses the bond between the 3'-end of DNA and stalled DNA topoisomerase 1. the importance of TDP1 is highlighted by its association with the human genetic disease spinocerebellar ataxia with axonal neuropathy. TDP1 comprises of a highly conserved C-terminus phosphodiesterase domain and a less conserved N-terminus tail. the importance of the N-terminus domain was suggested by its interaction with Lig3alpha. Here we show that this interaction is promoted by serine 81 that is located within a putative S/TQ site in the N-terminus domain of TDP1. Although mutation of serine 81 to alanine had no impact on TDP1 activity in vitro and had little impact on the ability of TDP1 to mediate the rapid repair of CPT- or IR-induced DNA breaks in vivo, it led to marked reduction of protein stability. Moreover, it reduced the ability of TDP1 to promote cell survival following genotoxic stress. Together, our findings highlight a novel mechanism for regulating TDP1 function in mammalian cells that is not directly related to its enzymatic activity. PMID- 20009513 TI - Universal immunization of infants against Neisseria meningitidis: addressing the remaining unmet medical need in the prevention of meningitis and septicemia. AB - Effective immunization policies have markedly decreased the incidence of many lethal infectious diseases of childhood, including diphtheria, pertussis, and poliomyelitis, among others. In industrialized countries, relatively recent success in combating meningitis and sepsis has come with the implementation of universal immunization of infants against Haemophilus influenzae type b (Hib) and Streptococcus pneumoniae. These universal immunization programs have reduced disease incidence and related deaths by more than 90%. Thus 2 out of the 3 major bacterial causes of invasive disease in children have now been controlled, leaving Neisseria meningitidis as the obvious next target. Currently, mortality attributable to invasive meningococcal disease remains at a level consistent with that of several other major vaccine-preventable infections prior to the implementation of immunization strategies. Unlike Hib and pneumococcus, US immunization policy against invasive meningococcal disease currently focuses on adolescents, a strategy that has been notably less than successful given that the highest incidence of invasive meningococcal disease occurs early in the first year of life. Development of safe and effective vaccines that broadly protect infants against disease caused by N. meningitidis is the next logical step in the effort to prevent bacterial meningitis and sepsis. Their universal use in infancy would follow a well-established and notably successful path. PMID- 20009514 TI - Radical-induced cell death1 and similar to RCD one1 and the stress-induced morphogenetic response. PMID- 20009515 TI - European Malaria Vaccine Initiative: portfolio and perspectives for the future. AB - Over the past ten years, EMVI has continually strived to maintain its main goal of accelerating the development of candidate malaria vaccines by facilitating the translational gap between promising experimental malaria vaccines and subsequent clinical trials in Europe and in Africa. To date, EMVI has funded approximately ten vaccine formulations (antigen-adjuvant combination) by developing GMP materials and sponsoring subsequent human clinical trials. In recent years EMVI's role has expanded into harmonization activities relevant to malaria vaccine development as well as making contributions to global coordination efforts in the field of malaria vaccine research and development (R&D). In the next five years, EMVI will be coordinating the European Network of Vaccine Research and Development, an European Commission supported action implementing a vaccine development infrastructure for Europe. By stimulating collaboration, cooperation, networking and joint integrated activities across various fields of research and diseases, and by facilitating the federation of research infrastructures, EMVI is acting today as a catalyst for tomorrow's vaccines. PMID- 20009517 TI - Impact of wastage on single and multi-dose vaccine vials: Implications for introducing pneumococcal vaccines in developing countries. AB - INTRODUCTION: Pneumococcal conjugate vaccines are expensive relative to those in the EPI systems of low-income countries. The current single-dose presentation costs more to store in the cold chain relative to multi-dose presentations but also has lower wastage rates. It is, therefore, important to determine the optimal balance of vial size and storage costs after adjusting for wastage. OBJECTIVES: To project the cost implications of wastage when vaccine wastage rates vary across vial sizes using country specific wastage data. RESULTS: Only 19 (26%) of 72 GAVI eligible countries had analyzable wastage data at WHO/HQ. The median wastage rates for single, 2- and 10-dose vials were 5%, 7% and 10% respectively. However wastage varied between 1%-10%, 1%-27% and 4%-44% for single, 2- and 10-dose vials respectively. The increased variance for multi-dose vial wastage implied wastage costs potentially greater than the savings realized from lower storage volumes. METHODS: For each potential vial size, we estimated cold chain costs and the cost of wasted vaccine doses using country level wastage data and projections of the price per dose of vaccine and cold chain storage. CONCLUSIONS: The optimal vial-size for PCV is dependent upon country specific wastage rates but few countries have these data. There may be a role for both single and multi-dose vials that is best determined by local management and storage capacities making local wastage data critical. Without effective wastage monitoring and control there is a risk that wastage costs will possibly exceed the savings from multi-dose vials' lower storage costs. PMID- 20009516 TI - Conserved developmental alternative splicing of muscleblind-like (MBNL) transcripts regulates MBNL localization and activity. AB - Muscleblind-like (MBNL) proteins have been shown to regulate pre-mRNA alternative splicing, and MBNL1 has been implicated in regulating fetal-to-adult transitions in alternative splicing in the heart. MBNL1 is highly conserved, exhibiting more than 95% identity at the amino acid level between birds and mammals. To investigate MBNL1 expression during embryonic heart development, we examined MBNL1 transcript and protein expression in the embryonic chicken heart from the formation of the primitive heart tube through cardiac morphogenesis (embryonic days 1.5 through 8). MBNL1 transcript levels remained steady throughout these stages, whereas MBNL1 protein levels increased and exhibited a shift in isoforms. MBNL1 has several alternatively spliced exons. Using RT-PCR, we determined that the inclusion of one of these, exon 5, decreases dramatically during cardiac morphogenesis. This developmental transition is conserved in mice. Functional analyses of MBNL1 isoforms containing or lacking exon 5-encoded sequences revealed that exon 5 is important for the regulation of the subcellular localization, RNA binding affinity, and alternative splicing activity of MBNL1 proteins. A second MBNL protein, MBNL2, is also expressed in the embryonic heart. We found that MBNL2 exon 5, which is paralogous to MBNL1 exon 5, is similarly regulated during embryonic heart development. Analysis of MBNL1 and MBNL2 transcripts in several embryonic tissues in chicken and mouse indicate that exon 5 alternative splicing is highly conserved and tissue-specific. Thus, we propose that conserved developmental stage- and tissue-specific alternative splicing of MBNL transcripts is an important mechanism by which MBNL activity is regulated during embryonic development. PMID- 20009518 TI - Thigmomorphogenesis in Solanum lycopersicum: morphological and biochemical responses in stem after mechanical stimulation. PMID- 20009519 TI - Molecular characterization of human rotavirus vaccine strain CDC-9 during sequential passages in Vero cells. AB - We have developed several candidate human rotavirus vaccine strains with common serotypes via adaptation in Vero cells, adhering to the Good Laboratory Practice (GLP) guidelines. We sequenced the entire genome of a G1P[8] strain CDC-9 in original stool and passaged materials from Vero cells and examined its genetic relatedness to the prototype human rotavirus KU and other strains. With the exception of VP3 gene which was closely related to that of strain DS-1, the culture-adapted CDC-9 strain shared moderate to high nt (range 83.4%-95.1%) and deduced aa (range 81.3%-97.9%) sequence identities with the KU and other G1P[8] strains. Alignments of the deduced aa sequences of 11 gene segments of the wild type and culture-adapted CDC-9 showed complete sequence identity in genes encoding NSP2, NSP3, NSP4, VP1, VP2, VP3 and VP7, a single aa change in genes coding for NSP1, NSP5 and VP6 and several scattered aa changes in the VP4 gene during the passage in Vero cells. Two of the VP4 aa substitutions (385 and 388) are within sites associated with neutralization resistant mutants selected by cross-reactive monoclonal antibodies. Although some sequence changes were evident, we do not know if these changes contribute to the possible attenuation of this strain. Further testing of this vaccine strain in clinical trials is justified. PMID- 20009520 TI - Functional diversity of the plant glycine-rich proteins superfamily. AB - The first plant glycine-rich proteins (GRPs) have been isolated more than 20 years ago based on their specific expression pattern and/or modulation by several biotic and abiotic factors. This superfamily is characterized by the presence of a glycine-rich domain arranged in (Gly)(n)-X repeats. The presence of additional motifs, as well as the nature of the glycine repeats, groups them in different classes. The diversity in structure as well as in expression pattern, modulation and sub-cellular localization have always indicated that these proteins, although classified as members of the same superfamily, would perform different functions in planta. Only now, two decades later, with the first functional characterizations of plant GRPs their involvement in diverse biological and biochemical processes are being uncovered. Here, we review the so far ascribed functions of plant GRPs. PMID- 20009521 TI - Brassinosteroid negatively regulates jasmonate inhibition of root growth in Arabidopsis. PMID- 20009522 TI - Immunological persistence in 4-6 and 7-9 year olds previously vaccinated in infancy with hexavalent DTPa-HBV-IPV/Hib. AB - BACKGROUND: The combined diphtheria-tetanus-pertussis-hepatitis B-inactivated poliomyelitis-Haemophilus influenzae conjugate vaccine (DTP a-HBV-IPV/Hib, Infanrix Hexa() GlaxoSmithKline Biologicals, Rixensart, Belgium) is the only hexavalent vaccine currently licensed for primary and booster vaccination of infants and provides simultaneous protection against six major diseases of childhood. The persistence of the immune response in children aged 4-6 and 7-9 years of age previously vaccinated with four doses of DTP a-HBV-IPV/Hib vaccine was assessed (www.clinicaltrials.gov.au 106744 NCT00356564 and 106745 NCT00335881). METHODS: A blood sample was collected from 403 children, all of whom had received 3-dose primary vaccination and a booster dose in the second year of life with DTP a-HBV-IPV/Hib, in previous clinical vaccine trials in Germany. RESULTS: Mean time from the fourth DTP a-HBV-IPV/Hib dose until serological follow-up ranged between 3.6 and 6.4 years. After the 4th DTP a-HBV IPV/Hib dose, in subjects who had not received additional booster doses, seroprotective antibody levels persisted up to 9 years of age in >/=90% of subjects for diphtheria, Hib and poliomyelitis, in 77.2% subjects for Hepatitis B and in 64.7% of subjects for tetanus. Anti-pertussis toxin antibodies remained detectable in no more than 38.2% of subjects. CONCLUSION: With the exception of PT , the combined DTP a-HBV-IPV/Hib induces long lasting immune response against all vaccine antigens. Falling seropositivity against PT over time supports the recommended administration of a pertussis booster dose in 5-6 year old children in Germany. PMID- 20009524 TI - PATH Malaria Vaccine Initiative (MVI): perspectives on the status of malaria vaccine development. PMID- 20009523 TI - Pin1 expression contributes to lung cancer: Prognosis and carcinogenesis. AB - Lung cancer remains the most common cause of death for malignancy in both men and women. Current therapies for NSCLC patients are inefficient due to the lack of diagnostic and therapeutic markers. The phospho-Ser/Thr-Pro specific prolyl isomerase Pin1 is overexpressed in many different cancers, including NSCLC, and may possibly be used as a target for cancer therapy. We identified 79 cases with the follow-up survival and investigated the clinical relevance of Pin1 expression in NSCLC patients. To validate the oncogenic potential of Pin1 in lung cells, we overexpressed Pin1 in Glc82 cells, and downregulated Pin1 by RNA interference in H1299 cells. The 5-year survival rate of the 79 patients was 54.6%. High expression of Pin1 correlated with poor survival by univariate analysis as well as by multivariate analysis, demonstrating that high expression of Pin1 was an independent prognostic factor. Consistent with the clinical findings, overexpression of Pin1 in Glc82 cells increased cell growth and colony formation and tumorigenicity in nude mice including cell migration, invasion. To further validate the role of Pin1 in lung cancer carcinogenesis, lentivirus-mediated siRNA targeting of Pin1 resulted in the stable suppression of both cell growth, anchorage-independent growth in soft agar and tumorigenic including cell migration, invasion in H1299 cells. Pin1 expression may be an unfavorable prognostic factor in patients of NSCLC patients, and these results indicate that Pin1 may have a role in tumor development and metastasis and thus could serve as a novel target for treatment of NSCLC. PMID- 20009525 TI - Computational approaches for understanding the evolution of DNA methylation in animals. AB - DNA methylation is implicated in modulating gene transcription in a wide array of taxa. However, a complete understanding of the specific functions of DNA methylation in diverse organisms requires study of the evolutionary patterns of genome methylation. Unfortunately, investigating DNA methylation through empirical means remains challenging due to the transient nature of DNA methylation, the paucity of model systems in which to study genome methylation, and the costs associated with experimental methodology. Here we review how computational methods can be used to complement experimental approaches to further our understanding of DNA methylation in animals. For instance, comparative analyses of the molecular machinery involved in DNA methylation have been informative in revealing the dynamics of genome methylation in many organisms. In addition, analyses of specific genomic signatures of DNA methylation have furthered our understanding of the patterns of methylation within species. Finally, insight into the role of DNA methylation has resulted from computational methods used to identify specific sets of methylated genes. We suggest that an understanding of the evolution of genomic DNA methylation can be most readily achieved by integrating computational and empirical methods. PMID- 20009526 TI - Current status of Plasmodium vivax vaccine. AB - From a total of 2.6 billion people at permanent risk of suffering malaria infection worldwide, 80-300 million experience Plasmodium vivax infections every year, with clinical manifestations ranging from asymptomatic to mild and chronic infection that in some cases lead to severe disease and death. The increasing P. vivax drug resistance and reports of severe and lethal cases, the relapsing parasite behavior and the existence of Plasmodium spp. co-infections must prompt more investment and greater efforts for the development of P. vivax vaccine. Currently there are only two P. vivax vaccine candidates being tested in clinical trials and few others are being assessed in preclinical studies which contrast with the numerous P. falciparum vaccines candidates under evaluation. The recent availability of the P. vivax genome and ongoing proteomic analysis are likely to accelerate P. vivax vaccine development. Recent development of human sporozoite challenge models would contribute to move clinical development forward and to identify mechanisms of immunity. PMID- 20009527 TI - Exploring the immunome: A brave new world for human vaccine development. PMID- 20009528 TI - Autophagy, ferritin and iron chelation. PMID- 20009530 TI - Early growth response 1 (EGR1): a gene with as many names as biological functions. AB - Although most researchers in biology tend to focus on very specific issues and questions about their preferred gene or pathway, sometimes we face situations in which nature presents us with a remarkable example of a gene with multiple functions. Since the discovery of the early growth response 1 (EGR1) gene in the mid eighties, several independent groups attributed its activation as an immediate early response gene to extracellular stimuli such as environmental cues, growth factors, irradiation and small molecules. Even twenty-plus years after its initial cloning and characterization, EGR1 continues to attract considerable attention among biological circles. Recently, work by Wagers and colleagues reported yet another novel and rather promising function for EGR1. They demonstrated that EGR1 is involved in regulating homeostasis of hematopoietic stem cells (HSC) by coordinating proliferation and migration. Importantly, this finding allows a conduit by which manipulation of this pathway will elucidate further understanding of the biology driving HSC mobilization and transplantation in a clinical setting. PMID- 20009529 TI - Axon-glial interactions at the Drosophila CNS midline. AB - The glia that reside at the midline of the Drosophila CNS are an important embryonic signaling center and also wrap the axons that cross the CNS. The development of the midline glia (MG) is characterized by migration, ensheathment, subdivision of axon commissures, apoptosis, and the extension of glial processes. All of these events are characterized by cell-cell contact between MG and adjacent neurons. Cell adhesion and signaling proteins that mediate different aspects of MG development and MG-neuron interactions have been identified. This provides a foundation for ultimately obtaining an integrated picture of how the MG assemble into a characteristic axonal support structure in the CNS. PMID- 20009531 TI - Cell migration and metastasis markers as targets of environmental pollutants and the Aryl hydrocarbon receptor. AB - During the last few years, several studies have pointed to a surprising link between environmental pollutants cellular signaling and important cell functions such as plasticity, adhesion and migration. This unexpected link could be related to endogenous functions of pollutants receptors that may be disrupted by environmental factors, which is supported by observations in invertebrate species. It could also reveal novel toxic end-points and mechanisms of those pollutants, such as teratogenesis and cancer metastasis that are highly relevant from a public health point of view. In the present short article, we will review our recent observations on the aryl hydrocarbon receptor and its new molecular and cellular targets. We identified HEF1/NEDD9/CAS-L, a multifunctional protein involved in integrin-based signaling as a transcriptional target of the receptor, and showed that its induction was critical for cell plasticity mediated by environmental pollutants. We will put our studies in perspective with other observations made by several groups. PMID- 20009532 TI - Hot-spot mutations in p110alpha of phosphatidylinositol 3-kinase (pI3K): differential interactions with the regulatory subunit p85 and with RAS. AB - The phosphatidylinositol 3-kinase (pI3K) signaling pathway is frequently upregulated in cancer. PIK3CA, the gene coding for the catalytic subunit p110alpha of PI3K, is mutated in about 12% of all human cancers. Most of these mutants are single amino acid substitutions that map to three positions (hot spots) in the helical or kinase domains of the enzyme. The mutant proteins show gain of enzymatic function, constitutively activate AKT signaling and induce oncogenic transformation in vitro and in animal model systems. We have shown previously that hot-spot mutations in the helical domain and kinase domain of the avian p110alpha have different requirements for interaction with the regulatory subunit p85 and with RAS-GTP. Here, we have carried out a genetic and biochemical analysis of these "hot-spot" mutations in human p110alpha. The present studies add support to the proposal that helical and kinase domain mutations in p110alpha trigger a gain of function by different molecular mechanisms. The gain of function induced by helical domain mutations requires interaction with RAS-Gtp. In contrast, the kinase domain mutation is active in the absence of RAS-Gtp binding, but depends on the interaction with p85. PMID- 20009533 TI - Mitochondrial protease AtFtsH4 protects ageing Arabidopsis rosettes against oxidative damage under short-day photoperiod. PMID- 20009534 TI - Larval midgut destruction in Drosophila: not dependent on caspases but suppressed by the loss of autophagy. AB - While most programmed cell death (PCD) in animal development is reliant upon the caspase-dependent apoptotic pathway and subsequent cleavage of caspase substrates, we found that PCD in Drosophila larval midgut occurs normally in the absence of the main components of the apoptotic machinery. However, when some of the components of the autophagic machinery were disrupted, midgut destruction was severely delayed. These studies demonstrate that Drosophila midgut PCD is executed by a novel mechanism where caspases are apparently dispensable, but that requires autophagy. PMID- 20009535 TI - In vivo subcellular targeting analysis validates a novel peroxisome targeting signal type 2 and the peroxisomal localization of two proteins with putative functions in defense in Arabidopsis. PMID- 20009536 TI - Epigenetic regulation of myogenesis. AB - Adult skeletal muscle provides a unique paradigm for studying stem to differentiated cell transitions. In response to environmental stress, quiescent muscle stem cells (satellite cells) are activated and proliferative, at which stage they can either differentiate and fuse to form new muscle fibers or alternatively self-renew and maintain the muscle stem cell reservoir. This multi step myogenic process is orchestrated by muscle regulatory proteins such as Pax3/Pax7 and members of the MyoD family of transcription factors. Findings published over the past few years have uncovered that epigenetic mechanisms critically repress, maintain or induce muscle-specific transcriptional programs during myogenesis. These studies are increasing our understanding of how muscle lineage-specific information encoded in chromatin merges with muscle regulatory factors to drive muscle stem cells through transitions during myogenesis. PMID- 20009537 TI - Epigenetic status determines germ cell meiotic commitment in embryonic and postnatal mammalian gonads. AB - The meiotic cell cycle is required for production of fertilization-competent gametes. Germ cell meiotic commitment requires expression of Stimulated by retinoic acid gene 8 (Stra8), which is transcriptionally activated by retinoic acid (RA). Meiotic suppression in embryonic male germ cells is believed to result from sex-specific differences in CYP26B1-catalyzed RA metabolism in the developing gonads. Here we show in mice that RA-induced Stra8 transcription is epigenetically controlled and requires a co-activator that binds proximal to the RA response elements (RAREs) in the Stra8 promoter. Embryonic male germ cells exposed in utero to the class I/II histone deacetylase (HDAC) inhibitor, trichostatin-A (TSA), show premature Stra8 activation and meiotic entry without altered Cyp26b1 expression. We also show that Stra8 expression is detectable and physiologically regulated in adult mouse ovaries. Further, oogenesis induction in adult females using TSA is associated with Stra8 activation, and these events are absent in mice deficient in the RA precursor vitamin A. Finally, all of the actions of TSA in premeiotic germ cells in vitro and in mouse ovaries in vivo can be reproduced with the small molecule HDAC inhibitor, suberoylanilide hydroxamic acid (SAHA). Thus, the ability of RA to transcriptionally induce expression of the meiosis-commitment gene, Stra8, is epigenetically controlled and this process involves a novel co-activator that functions upstream of the RAREs. These events not only coordinate the sex-specific timing of meiotic entry during embryogenesis, but also contribute to the regulation of oogenesis in adult female mammals. PMID- 20009538 TI - Localization of prion-destabilizing mutations in the N-terminal non-prion domain of Rnq1 in Saccharomyces cerevisiae. AB - [PIN(+)] is the prion form of Rnq1 in Saccharomyces cerevisiae and is necessary for the de novo induction of a second prion, [PSI(+)]. The function of Rnq1, however, is little understood. The limited availability of defective rnq1 alleles impedes the study of its structure-function relationship by genetic analysis. In this study, we isolated rnq1 mutants that are defective in the stable maintenance of the [PIN(+)] prion. Since there is no rnq1 phenotype available that is applicable to a direct selection or screening for loss-of-function rnq1 mutants, we took advantage of a prion inhibitory agent, Rnq1Delta100, to develop a color based genetic screen. Rnq1Delta100 eliminates the [PSI(+)] prion in the [PIN(+)] state but not in the [pin(-)] state. This allows us to find loss-of-[PIN(+)] rnq1 mutants as white [PSI(+)] colonies. Nine rnq1 mutants with single-amino-acid substitutions were defined. These mutations impaired the stable maintenance of [PIN(+)] and, as a consequence, were also partially defective in the de novo induction of [PSI(+)]. Interestingly, eight of the nine alleles were mapped to the N-terminal region of Rnq1, which is known as the non-prion domain preceding the asparagine and glutamine rich prion domain of Rnq1. Notably, overexpression of these rnq1 mutant proteins restored [PIN(+)] prion activity, suggesting that each of the rnq1 mutants was not completely inactive. These findings indicate that the N-terminal non-prion domain of Rnq1 harbors a potent activity to regulate the maintenance of the [PIN(+)] prion. PMID- 20009539 TI - Effects of combined inhibition of MEK and mTOR on downstream signaling and tumor growth in pancreatic cancer xenograft models. AB - The ERK and mTOR pathways show multiple interconnections that coordinate growth activation and the regulation of protein translation. Although drugs that target these pathways appear to have limited anti-cancer effects as single agents, we hypothesized that the monotherapy anticancer efficacy of these agents could be enhanced by their combination. The MEK inhibitor AZD6244 (ARRY-142886) and the mTOR inhibitor rapamycin were tested as single agents and in combination, using BxPC-3 and MIA PaCa-2 pancreatic cancer models in vivo. In both models, S6 ribosomal protein was almost completely inhibited with combined treatment, but only partially inhibited with the single agents. In addition, 48 h treatment with the drug combination produced greater apoptosis, revealed by caspase 3 cleavage, and growth inhibition measured using bromodeoxyuridine incorporation, compared to the single agents. AZD6244 but not rapamycin exhibited a significant anti angiogenic effect, as shown by tumor VEGF ELISA assay and CD31 analysis. Plasma and tumor pharmacokinetic analyses indicated that AZD6244 accumulates in tumor tissue at concentrations that produce target inhibition and cell cycle arrest in vitro. In chronic dosing experiments, the drug combination was well tolerated, and showed greater growth inhibition compared to the single agents. These results are consistent with the hypothesis that ERK and mTOR signaling interact at multiple levels to regulate tumor growth in vivo, and support the testing of MEK plus mTOR inhibitor combinations in pancreatic cancer patients. PMID- 20009540 TI - Self-reported history of vaccination and disease and immunity against hepatitis A, hepatitis B, tetanus, diphtheria and varicella among Spanish military recruits. AB - This study aims to evaluate the immune status against hepatitis A, hepatitis B, tetanus, diphtheria and varicella in military recruits and the validity of self reporting of their disease and vaccination history. A total of 226 participants were studied (mean age, 20.2 years; SD 1.7). 10.4% presented antibodies to hepatitis A, 78.3% to hepatitis B, 94.2% to tetanus, 77.4% to diphtheria and 81.9% to varicella. The relationship between self-reporting of vaccination history and seroprotection showed a high Positive Predictive Value for tetanus (98.8%) and a high Negative Predictive Value for hepatitis A (91%). Hepatitis A vaccination and serology testing for varicella and Hepatitis B on joining the Spanish armed forces are recommended. PMID- 20009541 TI - The overexpression of ERCC-1 is involved in the resistance of lung cancer cells to cetuximab combined with DDP. AB - Cetuximab, an antibody against epidermal growth factor receptor, has been approved for the treatment of colorectal carcinoma and head and neck squamous cell carcinoma. There is increasing evidence that cetuximab can reverse the resistance to irinotecan (CPT-11) and oxaliplatin. Since cisplatin (DDP) is a widely used chemotherapeutics this study examined whether cetuximab could reverse the resistance to DDP. Combined treatment with DDP and cetuximab resulted in an increase in the cytotoxicity of DDP in a DDP-sensitive lung cancer cell line (A549), but not in a DDP-resistant derivative (A549/DDP). Meantime, DDP activated the EGFR pathway in A549 cells but not in A549/DDP cells in a ligand-independent fashion. After the expression of excision repair cross-complementation group 1 (ERCC-1) protein was inhibited by small interfering RNA (siRNA), the potential of cetuximab to enhance DDP-mediated cytotoxicity was restored in A549/DDP cells. These data suggested that ERCC-1 was involved in the resistance of cetuximab combined with DDP as overexpression of ERCC-1 prohibits the activation of EGFR pathway, which would facilitate the preselection of lung cancer patients for the treatment of cetuximab combined with DDP. PMID- 20009542 TI - CT response assessment combining reduction in both size and arterial phase density correlates with time to progression in metastatic renal cancer patients treated with targeted therapies. AB - BACKGROUND: Response assessment is critical in evaluating effectiveness of anticancer treatment. Tyrosine kinase inhibitors (TKIs) in renal cell carcinoma (RCC) are associated with significant clinical benefit but may not result in significant tumor size reduction. Thus standard size-based response assessment with RECIST is insensitive, resulting in low response rates which do not reflect disease control measured by time to progression. We compared the use of combined size and density response criteria with standard size based criteria in metastatic RCC patients treated with TKI's. RESULTS: Partial response (PR) and stable disease (SD) defined by modified criteria successfully identified patients with a long TTP (448 days) or short TTP (89 days) respectively (p = 0.002). Neither RECIST nor standard Choi criteria successfully discriminated between patients having a short or long clinical benefit. PATIENTS AND METHODS: CT scans from 32 patients with metastatic RCC treated with either sunitinib (18) or cediranib (14) were assessed. Twelve patients were excluded from the analysis as ten had non-contrast enhanced scans due to renal impairment and two stopped treatment due to toxicity. Scans from 20 evaluable patients at baseline and 12 w on treatment were assessed using RECIST, Choi and modified criteria in which both a 10% decrease in size and 15% decrease in density were required to define a partial response (PR). Response assessment performed using each of the three methods was compared with time to disease progression (TTP) defined by RECIST using Kaplan-Meier statistics and Log-rank test with significance at 5%. CONCLUSION: A combined reduction in both size and arterial phase density of RCC metastases treated with TKIs correlates with TTP. RECIST and standard Choi criteria appear inferior. PMID- 20009544 TI - PtdIns(3,5)P2 and autophagy in mouse models of neurodegeneration. PMID- 20009545 TI - Plant defensins: defense, development and application. AB - Plant defensins are small, highly stable, cysteine-rich peptides that constitute a part of the innate immune system primarily directed against fungal pathogens. Biological activities reported for plant defensins include antifungal activity, antibacterial activity, proteinase inhibitory activity and insect amylase inhibitory activity. Plant defensins have been shown to inhibit infectious diseases of humans and to induce apoptosis in a human pathogen. Transgenic plants overexpressing defensins are strongly resistant to fungal pathogens. Based on recent studies, some plant defensins are not merely toxic to microbes but also have roles in regulating plant growth and development. PMID- 20009546 TI - The insect-trapping rim of Nepenthes pitchers: surface structure and function. AB - Carnivorous pitcher plants of the genus Nepenthes capture prey with a pitfall trap that relies on a micro-structured, slippery surface. The upper pitcher rim (peristome) is fully wettable and causes insects to slip by aquaplaning on a thin water film. The high wettability of the peristome is probably achieved by a combination of hydrophilic surface chemistry, surface roughness and the presence of hygroscopic nectar. Insect foot attachment could be prevented by the delayed drainage of the thin water film between the adhesive pad and the surface. Drainage should be faster for insects with a hairy adhesive system; however, they slip equally on the wet peristome. Therefore the stability of the water film against dewetting appears to be the key factor for aquaplaning. New experimental techniques may help to clarify the detailed function of the pitcher plant peristome and to explore its potential for biomimetic applications. PMID- 20009548 TI - Nibbling away at synaptic development. AB - Beyond its role as a response to starvation, autophagy has been increasingly implicated as part of the normal mechanisms regulating growth and remodeling of various cells and tissues during development. In recently published work we demonstrate that autophagy promotes synaptic development of the Drosophila larval neuromuscular junction (NMJ). We find that autophagy acts by downregulating an E3 ubiquitin ligase, Highwire (Hiw), which limits NMJ growth via a MAPKKK pathway. A similar role for autophagy in the synaptic remodeling that occurs during learning and memory remains an intriguing possibility. PMID- 20009549 TI - Targeting Beclin 1 for viral subversion of macroautophagy. AB - We have recently characterized that influenza A virus blocks autophagosome degradation via its matrix protein 2. Matrix protein 2 seems to achieve this macroautophagy inhibition not by its well-characterized proton channel function, but possibly due to its binding to Atg6/Beclin 1, thereby enhancing the death of its host cell. Here we discuss several viruses that now have been described to compromise macroautophagy via binding to Atg6/Beclin 1 with different outcomes for their replication, and how interaction with one and the same protein could inhibit autophagosome generation or degradation. PMID- 20009550 TI - Ethylene stimulation of latex production in Hevea brasiliensis. AB - Rubber tree (Hevea brasiliensis) is an important industrial crop for natural rubber production. Ethylene, as a stimulant of latex production in H. brasiliensis, has been widely used in commercial latex production. However, the mechanism of ethylene action are not completely elucidated, especially in molecular aspect. Here, we focus on the molecular biological progression of ethylene stimulation of latex production. Our data and all previous information showed ethylene had little direct effect on accelerating rubber biosynthesis. The prolonged latex flow and acceleration of sucrose metabolism by ethylene may be the main reasons for the stimulation of latex yield by ethylene. PMID- 20009551 TI - A variety of regulatory mechanisms are involved in the nitrogen-dependent modulation of the nodule organogenesis program in legume roots. AB - Combined nitrogen (N) sources are known to strongly affect initiation, development and functioning of Nitrogen-Fixing-Nodules whose formation is triggered by lipochitin-oligosaccharide signals secreted in the rhizospere by the Rhizobium partner. The rapid effects of N supply on nodule initiation have been mainly described when N sources are present at the moment of Rhizobium inoculation or purified Nod Factors addition. We recently reported that high ammonium nitrate growth conditions might also strongly affect the nodulation competence of Lotus japonicus plants, prior to the Rhizobium inoculation. This is a long-term effect, which suggests a change of the general nutritional status as the signal controlling the reduced nodulation capacities. The mechanisms underlying these inhibitory pathways are apparently different and the identification of the molecular actors involved may provide new insights into the linkage between N environmental changes and root organogenesis programs. PMID- 20009552 TI - Exploring the structural requirements for jasmonates and related compounds as novel plant growth regulators: a current computational perspective. AB - Jasmonates and related compounds have been highlighted recently in the field of plant physiology and plant molecular biology due to their significant regulatory roles in the signaling pathway for the diverse aspects of plant development and survival. Though a considerable amount of studies concerning their biological effects in different plants have been widely reported, the molecular details of the signaling mechanism are still poorly understood. This review sheds new light on the structural requirements for the bioactivity/property of jasmonic acid derivatives in current computational perspective, which differs from previous research that mainly focus on their biological evaluation, gene and metabolic regulation and the enzymes in their biosynthesis. The computational results may contribute to further understanding the mechanism of drug-receptor interactions in their signaling pathway and designing novel plant growth regulators as high effective ecological pesticides. PMID- 20009553 TI - Proline accumulation in plants: not only stress. AB - In addition to its role in protein synthesis and the plant cells' response to environmental stresses, circumstantial evidence suggest that proline may also play a role in flowering and development both as a metabolite and as a signal molecule. Although there is a growing consensus that proline is of special importance throughout the reproductive phase (from flower transition to seed development) a general agreement on the molecular and genetic mechanisms proline is involved in, is yet to be established. In this paper we shall review and critically discuss most of the evidence supporting a role for proline in plant development, paying special attention to the recently reported role of proline in flower transition. PMID- 20009555 TI - Plant PRA plays an important role in intracellular vesicular trafficking between compartments as GDF. AB - Rab GTPases like Ras-related monomeric GTPases are well known to regulate intracellular vesicle trafficking by cycling between membrane-bound and cytosolic states. The functions of these proteins are controlled by upstream regulators and downstream effectors. Ypt/Rabs transmit signals to downstream effectors in a GTP dependent manner. GDP-bound Rab proteins are extracted from their target membrane by cytosolic proteins known as GDP dissociation inhibitors (GDIs), and the Rab GTPase is recruited to the membrane compartment following dissociation from the GDI by GDI displacement factor (GDF). Now, we're going to discuss the role of plant PRA concerted with Rab and GDI proteins by recycling Rab between membrane and cytosol for intracellular trafficking of cargo proteins. PMID- 20009554 TI - Arabidopsis proline-rich extensin-like receptor kinase 4 modulates the early event toward abscisic acid response in root tip growth. AB - Exogenous application of plant hormone abscisic acid (ABA) can inhibit root growth. We recently reported that the proline-rich extensin-like receptor kinase 4 (PERK4) functions at an early stage of ABA signaling to inhibit primary root cell elongation by perturbing Ca2+ homeostasis.(1) Transcription analysis indicated that PERK4 modulates the expression of the genes related to cell elongation and ABA signaling in root growth, such as polygalacturonases, AtExt1, AtMYC2 and ABR1. Under ABA treatment, the transcript level of ZAT10, a Ca(2+) responsive gene, increased in perk4 plants compared to that of wild-type. Based on both present data and the previous evidence, we propose a probable model for PERK4-mediated ABA-regulated primary root cell growth. PMID- 20009556 TI - Redistribution of boron in leaves reduces boron toxicity. AB - High soil boron (B) concentrations lead to the accumulation of B in leaves, causing the development of necrotic regions in leaf tips and margins, gradually extending back along the leaf. Plants vary considerably in their tolerance to B toxicity, and it was recently discovered that one of the tolerance mechanisms involved extrusion of B from the root. Expression of a gene encoding a root B efflux transporter was shown to be much higher in tolerant cultivars. In our current research we have shown that the same gene is also upregulated in leaves. However, unlike in the root, the increased activity of the B efflux transporter in the leaves cannot reduce the tissue B concentration. Instead, we have shown that in tolerant cultivars, these transporters redistribute B from the intracellular phase where it is toxic, into the apoplast which is much less sensitive to B. These results provide an explanation of why different cultivars with the same leaf B concentrations can show markedly different toxicity symptoms. We have also shown that rain can remove a large proportion of leaf B, leading to significant improvements of growth of both leaves and roots. PMID- 20009557 TI - Expression of ACBP4 and ACBP5 proteins is modulated by light in Arabidopsis. AB - In our recent paper in Plant Physiology and Biochemistry, we reported that the mRNAs encoding Arabidopsis thaliana cytosolic acyl-CoA-binding proteins, ACBP4 and ACBP5, but not ACBP6, are modulated by light/dark cycling. The pattern of circadian-regulated expression in ACBP4 and ACBP5 mRNAs resembles that of FAD7 which encodes omega-3-fatty acid desaturase, an enzyme involved in plastidial fatty acid biosynthesis. Recombinant ACBP4 and ACBP5 proteins were observed to bind oleoyl-CoA ester comparably better than recombinant ACBP6, suggesting that ACBP4 and ACBP5 are promising candidates in the trafficking of oleoyl-CoA from the plastids to the endoplasmic reticulum (ER) for the biosynthesis of non plastidial membrane lipids. By western blot analyses using the ACBP4 and ACBP5 specific antibodies, we show herein that the levels of ACBP4 and ACBP5 proteins peak at the end of the light period, further demonstrating that they, like their corresponding mRNAs, are tightly controlled by light to satisfy demands of lipids in plant cells. PMID- 20009558 TI - Serine/threonine protein phosphatases type 2A and their roles in stress signaling. AB - Serine/threonine protein phosphatases are ubiquitous enzymes in all eukaryotes but many of their physiological roles in plants remain unknown. The available results have demonstrated critical functions for these enzymes in the regulation of adaptive stress responses, and recent studies have directed attention to the functional roles of Ser/Thr phosphatases type 2A (PP2A) as components of stress signaling pathways. This review is focused primarily on plant PP2As and their participation in the control of biotic and abiotic stress responses. PMID- 20009559 TI - Unique status of NIA2 in nitrate assimilation: NIA2 expression is promoted by HY5/HYH and inhibited by PIF4. AB - Light perceived by phytochromes will induce genes of nitrogen assimilation, however, transducing components in the signaling cascades to these genes are hardly known. Recently the bZIP transcription factors HY5 (LONG HYPOCOTYL5) and HYH (HOMOLOG OF HY5) were identified as positive regulators in light activation of NIA2 (nitrate reductase 2). The bHLH transcription factor PIF4 (PHYTOCHROME INTERACTING FACTOR 4) was revealed as an inhibitor of NIA2 expression. In contrast to NIA2, expression of other genes of nitrogen assimilation, NRT1.1 (dual-affinity nitrate transporter 1.1), NIA1 (nitrate reductase 1), NIR (nitrite reductase), GLN2 (glutamine synthetase 2) and GLU1 (glutamate synthase 1) were not promoted by HY5/HYH or inhibited by PIF4. NIA2 as the outstanding gene of nitrate assimilation regarding HY5/HYH and PIFs may have evolved in connection with the cytosolic leaf localization of nitrate reductase, and adverse effects of the products, nitrite, nitric oxide and active oxygen species formed by the enzyme. PMID- 20009560 TI - Autophagy in nuclear receptor PPARgamma-deficient mouse prostatic carcinogenesis. AB - Peroxisome proliferator-activated receptor-gamma (PPARgamma) is a major modulator of cellular lipid metabolism and organelle differentiation. To understand whether autophagy is involved in the processes of dysregulated fatty acid oxidation and induced oxidative stress accompanying prostatic carcinogenesis, we characterized in vitro and in vivo models of PPARgamma- and PPARgamma2-deficiency in mouse prostate epithelia. Autophagy accompanied the altered cellular proliferation and de-differentiation that resulted in PPARgamma-/gamma2-deficient mouse prostatic intraepithelial neoplasia (mPIN). Electron microscopy showed accumulated defective lysosomes and autophagic vacuoles in PPARgamma-/gamma2-deficient cells, suggestive of autophagy. Gene expression profiling indicated a major dysregulation of cell cycle control and metabolic signaling networks related to peroxisomal, mitochondrial and lysosomal maturation, lipid oxidation and degradation. Further, the putative autophagic phenotypes of PPARgamma-null cells could be rescued by re-expression of either the PPARgamma1 or -gamma2 isoform. Our paper examines the links between autophagy and PPARgamma-related subcellular and histopathological changes taking place during murine prostatic carcinogenesis. PMID- 20009561 TI - Autophagy dysfunction and ubiquitin-positive protein aggregates in Dictyostelium cells lacking Vmp1. AB - Ubiquitin-positive protein aggregates are a hallmark of many degenerative diseases. Their presence can be induced by dysfunction in protein degradation pathways such as proteasome and autophagy. We now report several lines of evidence suggesting a defect in autophagy in Dictyostelium cells lacking Vmp1 (vacuole membrane protein 1), an endoplasmic reticulum (ER)-resident protein involved in pathological processes such as cancer and pancreatitis. vmp1- null cells are unable to survive starvation or undergo autophagic cell death under the appropriate inductive signals. Moreover, confocal studies using the autophagy marker Atg8 and previous transmission electron microscopy analysis showed defects in autophagosome formation. Although Vmp1 is localized in the ER, we found colocalization with Atg8 suggesting a contribution of both Vmp1 and ER in autophagosome biogenesis or maturation. Interestingly, vmp1- mutant cells showed accumulation of huge ubiquitin-positive protein aggregates containing the autophagy marker GFP-Atg8 and the putative Dictyostelium p62 homologue as described in many degenerative human diseases. The analysis of other Dictyostelium autophagic mutants (atg1-, atg5-, atg6-, atg7- and atg8-) showed a correlation in the severity of their corresponding phenotypes and the presence of ubiquitin-positive protein aggregates suggesting that the deleterious effects associated with development of these aggregates might contribute to the complex phenotypes observed in autophagy deficient mutants. Our results suggest that Vmp1 is required for the clearance of these ubiquitinated protein aggregates through autophagy and highlight a potential role for Vmp1 in protein-aggregation diseases. PMID- 20009562 TI - Presence of yeasts in floral nectar is consistent with the hypothesis of microbial-mediated signaling in plant-pollinator interactions. AB - Olfactory floral signals are significant factors in plant-pollinator mutualisms. Recently, unusual fermentation odors have been described in the nectar and flowers of some species. Since yeasts are common inhabitants of many angiosperms nectars, this raises the possibility that nectar yeasts may act as causal agents of fermentation odors in flowers and, therefore, as possible intermediate agents in plant signaling to pollinators. A recent field study has reported that nectar yeasts were quite frequent in floral nectar across three different regions in Europe and America, where they reached high densities (up to 10(5) cells/mm(3)). Yeast incidence in floral nectar differed widely across plant host species in all sampling sites. A detailed study currently in progress on one of the species surveyed in that study (Helleborus foetidus, Ranunculaceae) has detected that, in addition to interespecific differences in yeast incidence, there is also a strong component of variance in yeast abundance that takes place at the subindividual level (among flowers of the same plant, among nectaries of the same flower). If yeast metabolism is eventually proved to contribute significantly to floral scent, then multilevel patchiness in the distribution of nectar yeasts (among species, among individuals within species, and among flowers and nectaries of the same individual) might contribute to concomitant multilevel variation in plant signaling and, eventually, also in pollination success, pollen flow and plant fitness. PMID- 20009563 TI - Extracellular ATP: a modulator of cell death and pathogen defense in plants. AB - Living organisms acquire or synthesize high energy molecules, which they frugally conserve and use to meet their cellular metabolic demands. Therefore, it is surprising that ATP, the most accessible and commonly utilized chemical energy carrier, is actively secreted to the extracellular matrix of cells. It is now becoming clear that in plants this extracellular ATP (eATP) is not wasted, but harnessed at the cell surface to signal across the plasma membrane of the secreting cell and neighboring cells to control gene expression and influence plant development. Identification of the gene/protein networks regulated by eATP mediated signaling should provide insight into the physiological roles of eATP in plants. By disrupting eATP-mediated signaling, we have identified pathogen defense genes as part of the eATP-regulated gene circuitry, leading us to the discovery that eATP is a negative regulator of pathogen defense in plants.(1) Previously, we reported that eATP is a key signal molecule that modulates programmed cell death in plants.(2) A complex picture is now emerging, in which eATP-mediated signaling cross-talks with signaling mediated by the major plant defense hormone, salicylic acid, in the regulation of pathogen defense and cell death. PMID- 20009564 TI - Alcohol exposure alters DNA methylation profiles in mouse embryos at early neurulation. AB - Alcohol exposure during development can cause variable neurofacial deficit and growth retardation known as fetal alcohol spectrum disorders (FASD). The mechanism underlying FASD is not fully understood. However, alcohol, which is known to affect methyl donor metabolism, may induce aberrant epigenetic changes contributing to FASD. Using a tightly controlled whole-embryo culture, we investigated the effect of alcohol exposure (88mM) at early embryonic neurulation on genome-wide DNA methylation and gene expression in the C57BL/6 mouse. The DNA methylation landscape around promoter CpG islands at early mouse development was analyzed using MeDIP (methylated DNA immunoprecipitation) coupled with microarray (MeDIP-chip). At early neurulation, genes associated with high CpG promoters (HCP) had a lower ratio of methylation but a greater ratio of expression. Alcohol induced alterations in DNA methylation were observed, particularly in genes on chromosomes 7, 10, and X; remarkably, a >10 fold increase in the number of genes with increased methylation on chromosomes 10 and X was observed in alcohol exposed embryos with a neural tube defect phenotype compared to embryos without a neural tube defect. Significant changes in methylation were seen in imprinted genes, genes known to play roles in cell cycle, growth, apoptosis, cancer, and in a large number of genes associated with olfaction. Altered methylation was associated with significant (p<0.01) changes in expression for 84 genes. Sequenom EpiTYPER DNA methylation analysis was used for validation of the MeDIP-chip data. Increased methylation of genes known to play a role in metabolism (Cyp4f13) and decreased methylation of genes associated with development (Nlgn3, Elavl2, Sox21 and Sim1), imprinting (Igf2r) and chromatin (Hist1h3d) was confirmed. In a mouse model for FASD, we show for the first time that alcohol exposure during early neurulation can induce aberrant changes in DNA methylation patterns with associated changes in gene expression, which together may contribute to the observed abnormal fetal development. PMID- 20009565 TI - Autophagy is induced by ischemic preconditioning in human livers formerly treated by chemotherapy to limit necrosis. AB - The effectiveness of ischemic preconditioning (IP) against hepatic ischemia/reperfusion injury during human liver surgery is linked to decreased apoptotic cell death as well as preservation of the ATP content in liver tissue. Overproduction of Bcl-2 is reported in preconditioned organs. In human liver biopsies exhibiting steatosis and/or vascular injuries (mainly peliosis) induced by chemotherapy, we find that the expression of Bcl-2 in centrolobular and peliotic areas colocalizes with the autophagy protein Beclin 1 in IP livers. Increased expression of phosphorylated Bcl-2 in preconditioned livers is associated with decreased immunoprecipitation of Beclin 1 and increased expression of LC3-II. The increased number of autophagic vacuoles seen by electron microscopy confirmed that IP could trigger autophagy in chemotherapy injured livers, probably to reduce the pro-inflammatory necrotic cell death of hepatocytes or endothelial cells and to increase ATP levels. Indeed, necrosis is less frequent (p = 0.04) in IP livers than in the others although no change in apoptosis as assessed by TUNEL assay or caspase-3, -8 and -9 expressions is observed. In conclusion, Bcl-2 and Beclin 1 could be major targets in the regulation of cell death during ischemia/reperfusion injury modulating autophagy to switch on/off necrosis and/or apoptosis. PMID- 20009566 TI - New frontiers in biomarker-targeted molecular imaging. PMID- 20009567 TI - Delocalization of gamma-tubulin due to increased solubility in human breast cancer cell lines. AB - The centrosome is the major organelle responsible for the nucleation and organization of microtubules into arrays. Recent studies demonstrate that microtubules can nucleate outside the centrosome. The molecular mechanisms controlling acentrosomal microtubule nucleation are currently poorly defined, and the function of this type of microtubule regulation in tumor cell biology is particularly unclear. Since microtubule nucleation is initiated by the gamma tubulin protein, we examined the regulation of gamma-tubulin in a panel of human breast tumor cell lines, ranging from non-tumorigenic to highly aggressive. We have identified a more dispersive subcellular localization of gamma-tubulin in aggressive breast cancer cell lines, while gamma-tubulin localization remains largely centrosomal in non-aggressive cell lines. Delocalization of gamma-tubulin occurs independently from changes in protein expression and is therefore regulated at the post-translational level. Subcellular fractionation revealed that tumor cell lines show an aberrantly increased release of gamma-tubulin into a soluble cytoplasmic fraction, with the most dramatic changes observed in tumor cell lines of greater aggressiveness. Extraction of soluble gamma-tubulin revealed acentrosomal incorporation of gamma-tubulin in cytoplasmic microtubules and along cell junctions. Moreover, acentrosomal delocalization of gamma-tubulin yielded resistance to colchicine-mediated microtubule collapse. These findings support a model where the solubility of gamma-tubulin can be altered through post translational modification and provides a new mechanism for microtubule dysregulation in breast cancer. Gamma-tubulin that is delocalized from the centrosome can still clearly be incorporated into filaments, and defines a novel mechanism for tumor cells to develop resistance to microtubule-targeted chemotherapies. PMID- 20009568 TI - Microbial alkaloid staurosporine induces formation of nanometer-wide membrane tubular extensions (cytonemes, membrane tethers) in human neutrophils. AB - In the present work, we demonstrate that microbial alkaloid staurosporine (STS) and Ro 31-8220, structurally related to STS protein kinase C inhibitor, caused development of membrane tubular extensions in human neutrophils upon adhesion to fibronectin-coated substrata. STS-induced tubular extensions interconnected neutrophils in a network and bound serum-opsonized bacteria Salmonella enterica serovar Typhimurium. The diameter of STS-induced extensions varied in the range 160-200 nm. The extensions were filled with cytoplasm and covered with membrane, as they included fluorescent cytoplasmic and lipid dyes. Neither protein kinase C inhibitors H-7 and bisindolylmaleimide VII, nor tyrosine protein kinase inhibitors tyrphostin AG 82 and genistein caused such extensions formation. Supposedly, STS induces membrane tubular extension formation promoting actin cytoskeleton depolymerization or affecting NO synthesis. PMID- 20009569 TI - Enterotoxigenic Bacteroides fragilis (ETBF)-mediated colitis in Min (Apc+/-) mice: a human commensal-based murine model of colon carcinogenesis. PMID- 20009570 TI - Double gaps along Shaker S4 demonstrate omega currents at three different closed states. AB - The aim of the present study was to investigate in detail how the voltage sensor in the Shaker potassium channel moves during the gating process. After the publication of the open channel structure from the crystallized K(V)AP channel in 2003, an alternative so-called "paddle" model was put forward in contrast to the existing helical screw model. The voltage sensor S4 contains 4 arginine residues relevant for gating, R1(362), R2(365), R3(368) and R4(371), each separated by 2 neutral residues. These charged residues coil as one of three threads on the S4 alpha-helix. Based on a previous finding that the mutation R1S leads to the so called omega leak current through a "gating-pore" in the closed state, we introduced gaps systematically along the arginine thread substituting long arginines by short serines. Mutations R2S or R3S did neither create transient nor steady leaks. The fact that the native residue A359, which is located three amino acids in front of R1, is a short one, motivated us to check its role. Mutation of A359 to arginine blocked the omega current in the R1S mutant indicating that the omega pore is occupied by A359 and R1. Introducing further double gaps (RR to SS) at sequential positions (0 + 1, 1 + 2, 2 + 3), produced clear leak currents which were remarkably stable over a wide voltage range. These leaks contradict that S4 would swing together with S3 in lipid according to the paddle hypothesis. Rather, our results show that during gating the S4 segment moves in 3 helical steps through a fixed pore formed by the channel protein. PMID- 20009571 TI - A translational end-run for a rare, genetically enigmatic tumor. PMID- 20009573 TI - Green chromatin dynamics in Zurich: meeting summary based on the European Workshop on Plant Chromatin 2009 in Zurich, Switzerland. AB - In September 2009, the 1(st) European Workshop on Plant Chromatin took place in Zurich, Switzerland. The workshop covered a variety of chromatin related topics, including the mechanisms of Polycomb group protein function, long-range interactions of regulatory elements, genome-wide reconfiguration of epigenetic marks during gametogenesis and protection of chromosomal ends by epigenetic marks. Some of the highlights of this meeting are summarized in this report. PMID- 20009572 TI - Roles of E3 ubiquitin ligases in cell adhesion and migration. AB - Recent studies have demonstrated that a number of E3 ubiquitin ligases, including Cbl, Smurf1, Smurf2, HDM2, BCA2, SCF(beta-TRCP) and XRNF185, play important roles in cell adhesion and migration. Cbl negatively regulates cell adhesion via alpha integrin and Rap1 and inhibits actin polymerization by ubiquitinating mDab1 and WAVE2. Smurf1 regulates cell migration through ubiquitination of RhoA, talin head domain and hPEM2, while Smurf2 ubiquitinates Smurf1, TGFbeta type I receptor and RaplB to modulate cell migration and adhesion. HDM2 negatively regulates cell migration by targeting NFAT (a transcription factor) for ubiquitination and degradation, while SCF(beta-TRCP) ubiquitinates Snail (a transcriptional repressor of E-cadherin) to inhibit cell migration. TRIM32 promotes cell migration through ubiquitination of Abl interactor 2 (Abi2), a tumor suppressor. RNF5 and XRNF185 modulate cell migration by ubiquitinating paxillin. Thus, these E3 ubiquitin ligases regulate cell adhesion and (or) migration through ubiquitination of their specific substrates. PMID- 20009574 TI - Matrix hyaluronan alters epidermal growth factor receptor-dependent cell morphology. AB - EGFR, a critical regulator of oncogenic signaling during cancer progression, is capable of integrating multireceptor signaling pathways that promote metastasis. EGFR is subject to regulatory cues from the extracellular matrix (ECM), of which hyaluronan (HA) is a major component. In mammary tumors, HA is deposited in the ECM where it functions in biomechanical support and modulates intracellular signaling. We utilized a 3D collagen system in which HA is either polymerized in collagen matrix or provided soluble in the media (sHA). Here we report that collagen-embedded HA (eHA) inhibits EGFR activation, filopodia formation and cell spreading on a collagen matrix. These findings demonstrate a novel role for eHA as a protective molecule when encountered in the collagen matrix during cancer progression. PMID- 20009576 TI - The hunt for hypoxia responsive natural antisense short interfering RNAs. AB - Small ribonucleic acids (smRNA) have been identified as important post transcriptional regulators of gene expression. One important class of smRNA implicated in stress responses are natural antisense short interfering RNA (natsiRNA). These natsiRNAs are generated from two overlapping mRNA that are processed into short-interfering RNAs and target complementary mRNA sequences for degradation. A feature of natsiRNA is the inverse regulation of gene expression that is observed after environmental or developmental stimuli. Genome-wide expression analysis of hypoxia treated Arabidopsis roots in combination with the examination of gene expression in plants defective in natsiRNA processing, was used to find putative natsiRNA regulated genes. The results indicate the potential involvement of natsiRNA in regulating lipid signaling in hypoxia. PMID- 20009575 TI - Targeting pericytes with a PDGF-B aptamer in human ovarian carcinoma models. AB - PURPOSE: On the basis of the known role of platelet-derived growth factor (PDGF) BB/PDGF receptor (PDGFR) beta in pericyte regulation, highly specific inhibitors of this target are needed. We tested the efficacy of a highly selective aptamer against PDGF-B with or without anti-VEGF therapy in ovarian cancer models. RESULTS: Bevacizumab inhibited tumor growth by 45% and 48% in the HeyA8 and SKOV3ip1 models, respectively. AX102 had minimal effect on the HeyA8 model, but increased tumor growth in the SKOV3ip1 model. However, bevacizumab plus AX102 was more effective than bevacizumab alone, and resulted in 76-88% inhibition of tumor growth in both models. A longitudinal study in the HeyA8 model using bioluminescence imaging showed that combination of bevacizumab, AX102 and paclitaxel caused tumor reduction by 65% (based on bioluminescence imaging). In the HeyA8 model, MVD and PCNA counts were significantly reduced in the bevacizumab treatment groups, and pericyte coverage was significantly decreased in the AX102 treatment groups. In the SKOV3ip1 model, MVD and PCNA was significantly reduced in the bevacizumab treatment group, and even lower in the bevacizumab and AX102 combination treatment group. EXPERIMENTAL DESIGN: The therapeutic efficacy of targeting endothelial cells (bevacizumab) and/or pericytes (PDGF-aptamer, AX102) was examined using HeyA8 and SKOV3ip1 orthotopic models of ovarian cancer metastasis. Following therapy, tumors were examined for microvessel density (MVD), proliferating cell nuclear antigen (PCNA) and vascular maturation (pericyte coverage). CONCLUSIONS: Dual targeting of endothelial cells and pericytes holds potential as an anti-vascular therapeutic approach in ovarian carcinoma. PMID- 20009577 TI - Cracking the code for compartment-specific dual functionality proteins in cancer: the case for CRKL. PMID- 20009578 TI - Model system takes us a step closer to efficacious imaging biomarkers of angiogenesis in head and neck cancer. PMID- 20009579 TI - Redox-regulation of protein import into chloroplasts and mitochondria: similarities and differences. AB - Redox signals play important roles in many developmental and metabolic processes, in particular in chloroplasts and mitochondria. Furthermore, redox reactions are crucial for protein folding via the formation of inter- or intramolecular disulfide bridges. Recently, redox signals were described to be additionally involved in regulation of protein import: in mitochondria, a disulfide relay system mediates retention of cystein-rich proteins in the intermembrane space by oxidizing them. Two essential proteins, the redox-activated receptor Mia40 and the sulfhydryl oxidase Erv1 participate in this pathway. In chloroplasts, it becomes apparent that protein import is affected by redox signals on both the outer and inner envelope: at the level of the Toc complex (translocon at the outer envelope of chloroplasts), the formation/reduction of disulfide bridges between the Toc components has a strong influence on import yield. Moreover, the stromal metabolic redox state seems to be sensed by the Tic complex (translocon at the inner envelope of chloroplasts) that is able to adjust translocation efficiency of a subgroup of redox-related preproteins accordingly. This review summarizes the current knowledge of these redox-regulatory pathways and focuses on similarities and differences between chloroplasts and mitochondria. PMID- 20009580 TI - Developmental strategies for evasion of Arf tumor suppression. PMID- 20009581 TI - Breast cancer metastasis driven by ErbB2 and 14-3-3zeta: A division of labor. AB - Metastasis remains the leading cause of cancer morbidity and mortality. ErbB2, a metastasis-promoting oncoprotein, is overexpressed in 50-60% of noninvasive ductal carcinoma in situ (DCIS). However, only 25% of invasive breast cancer (IBC) overexpress ErbB2, indicating that ErbB2 alone is not sufficient to drive metastasis and additional risk factors are necessary for the progression of ErbB2 overexpressing DCIS to IBC. A recent study published in Cancer Cell identified 14 3-3xi as a risk factor aiding the transition of ErbB2-overexpressing DCIS into IBC. Furthermore, the study elucidated molecular mechanisms by which ErbB2 and 14 3-3xi co-overexpression drives metastasis. Namely, ErbB2 promotes cell motility and migration via the activation of Src, while 14-3-3xi induces epithelial mesenchymal transition by activating TGFbeta pathway to reduce cell adhesion. On the other hand, two studies recently published in British Journal of Cancer and Oncogene provide mechanistic insight into how ErbB2 signalling is transduced via Src, focal adhesion kinase and Ste20-like kinase to regulate focal adhesion turnover and modulate cell motility and migration. Taken together, these studies reveal that metastasis engages a variety of players that must show team spirit to win the game of spreading. PMID- 20009582 TI - Estrogen contributes to non-pRb targeted HPV18 E7-caused cell proliferation and transformation. AB - Almost all of cervical carcinoma arises as a result of persistent infection with high-risk human papillomaviruses (HPVs) where E7 oncogene plays an important role. In addition, estrogen is a confirmed cofactor in HPV related cervical carcinoma working synergistically with E7. There are two pathways involved in the E7 carcinogenesis of cervical cancer, the pRb-dependent and pRb-independent pathway. In this study we analyzed that whether estrogen contributes to high-risk HPV E7 in cervical carcinogenesis via pRb-independent pathway or not. E7(DeltaRB), which can not bind to and degrade pRb protein, hence no transformation ability, worked synergistically with estrogen in cell proliferation and transformation. Estrogen combined with PTD-HPV18E7(DeltaRB) enhanced cell proliferation rate, induced genomic instability, including abnormal centrosome duplication and chromosomal instability, and exhibited malignant transformation with anchorage-independent growth. We also observed that, PTD HPV18E7(DeltaRB) can interact with c-Jun and c-Myc proteins, but this interaction was limited to the estrogen-treated cells where c-Jun and c-Myc were expressed highly and no such interaction was observed in estrogen-untreated cells where c Jun and c-Myc expression levels were low. In conclusion, estrogen can cooperate with E7 through a pRB-independent manner in cervical carcinogenesis. The functional interaction between E7 and c-Jun or between E7 and c-Myc could only be triggered when the c-Jun or c-Myc expression level reaches a certain threshold. PMID- 20009591 TI - Zimmermann-Laband syndrome in a child previously described with brachydactyly, extrahepatic biliary atresia, patent ductus arteriosus and seizures. PMID- 20009592 TI - Case of syndromic tufting enteropathy harbors SPINT2 mutation seen in congenital sodium diarrhea. PMID- 20009596 TI - Our right hand. PMID- 20009597 TI - Dental innovation: guided surgery in Europe. PMID- 20009598 TI - Piezoelectric internal sinus elevation (PISE) technique: a new method for internal sinus elevation. AB - This article reports the surgical procedure of a transalveolar approach for sinus floor elevation using piezoelectric surgery without a mallet, called piezoelectric internal sinus elevation (PISE) technique. PISE is the modified technique of the osteotome-mediated sinus floor elevation. Osteotome-mediated sinus floor elevation uses a mallet to break the sinus floor and to push sinus membrane up. Using this technique might possibly bring positional vertigo to the patient. PISE technique does not use a mallet during the procedure to break the sinus floor and to elevate the sinus membrane; therefore, it can reduce the possibility of postoperative positional vertigo. PMID- 20009599 TI - A technique for atraumatic extraction of teeth before immediate implant placement using implant drills. AB - PURPOSE: The purpose of this article is to present a minimally invasive technique using the implant drills to help extract teeth before the insertion of immediate implants. MATERIALS AND METHODS: Nine patients (7 women and 2 men) aged 24 to 60 years having root fractures and dental caries were included in this study. Inclusion criteria for the study were presence of at least 4 mm of bone beyond the root apex, the absence of acute signs of infection or inflammation in the treatment area, and the absence of systemic pathologies that would contraindicate bone healing around implants. Heavy smokers were not included in the study. Extraction was done after thinning the root walls by the help of the implant drills. After extraction, implant sites were prepared and implants were inserted. In 2 of the cases, the peri-implant bone defect was filled with autogenous bone graft. RESULTS: Healing progressed uneventfully in all 9 cases. The use of implant drills to thin the root walls provided atraumatic tooth extraction protecting the thin buccal bone. At second stage surgery, all implants were asymptomatic, immobile, and osseointegrated. The soft tissue anatomy was clinically acceptable in all patients. Radiographic examination of all 9 implants showed no peri-implant radiolucency at the end of a year. CONCLUSION: Successful osseointegration and complete bone healing were observed for all patients. The new extraction technique was found to be effective in immediate implant cases in order not to damage the thin plate of buccal bone. PMID- 20009600 TI - Incisive canal deflation for correct implant placement: case report. AB - AIM: This article is a case report of a patient in whom the prosthetic planning indicated the necessity of an incisive canal deflation for the correct installation of an implant that is to be osseointegrated. CASE REPORT: In the reopening phase after the bone graft installation, the incisive canal deflation (biopsy of its content) was done and titanium implants were installed with one of them invading the anatomical space occupied previously by the incisive canal. The biopsy analysis showed fragments of the incisive artery and nerve, which are responsible for the anterior upper-tooth pulp, the periodontium vascularization and the innervation. From the anastomosis present along with other structures allied with the absence of teeth in the region, there was no detriment to the patient caused by the deflation. CONCLUSION: Incisive canal deflation is a viable technique in implantology. It can permit ideal prosthetic planning with no detriment to the patient. PMID- 20009601 TI - Bone quality: a reality for the process of osseointegration. AB - This article subscribes to the initial condition of bone quality and its ultimate effect on the success of dental implant treatment. A high success rate for the preservation of the alveolar bone around oral implants is predicated on good bone quality. For implantologists, the classification of bone quality should be based on the key elements required for osseointegration. These include structural morphology and cellular characteristics such as monocytes/macrophages, mesenchymal progenitor cells, fibroblasts, osteoclasts, and cells associated with angiogenesis. PMID- 20009602 TI - Fabricating a nonrotational angulated abutment for a single-tooth prosthesis. AB - The fabrication of a custom waxed and cast nonrotational angulated abutment will be described. Most manufacturers of implant components fabricate angulated abutments for multiunit retrievable prostheses. However, they do not manufacture any components for a single unit that is screw retained. This article covers a step-by-step technique that was developed in an attempt to resolve this problem. PMID- 20009603 TI - CTX biochemical marker of bone metabolism. Is it a reliable predictor of bisphosphonate-associated osteonecrosis of the jaws after surgery? Part I: biological concepts with a review of the literature. AB - Biochemical markers of bone metabolism have been used in medicine to evaluate and provide treatment to patients with metabolic bone diseases, such as osteoporosis. Serum cross-linked C-telopeptide of type I collagen is a marker of osteoclast activity and is used to assess the level of bone resorption. Recently, in oral and maxillofacial surgery, it was proposed that the levels of serum cross-linked C-telopeptide of type I collagen may predict the subsequent risk of developing osteonecrosis of the jaws after oral surgery procedures for patients taking oral bisphosphonates. The astute clinician must critically review the scientific literature and must decide if biochemical markers of bone resorption are of benefit in managing the oral surgery patient on bisphosphonates. PMID- 20009604 TI - Comparison of prefabricated and custom-made bars used for implant-retained mandibular complete overdentures. AB - PURPOSE: To compare prefabricated and custom-made bars used for implant-retained mandibular complete overdentures. MATERIALS: Ten completely edentulous patients were selected for replacement with dental implants. Each patient received 2 (press-fit) dental implants, 1 implant on each side in the canine regions of the mandible. The implants were left submerged (unloaded) for a healing osseointegration period of 4 months. The patients were divided into 2 groups, 5 patients in each. Group I patients received maxillary conventional dentures and a mandibular overdenture retained by a cast bar. Group II patients received a maxillary conventional denture and a mandibular overdenture retained by a prefabricated bar. All patients were evaluated clinically and radiographically immediately after overdenture delivery and after 6, 12, and 18 months. RESULTS: There was more pronounced bone resorption in cast bar group more than the prefabricated bar group and minimal marginal bone loss in the group treated with prefabricated bar. CONCLUSION: The prefabricated bar overdentures showed less bone resorption distal to the implants in comparison with the cast bar implant retained overdentures, Both the gingival index and the plaque index score were significantly higher in the group treated with the cast bar retained overdenture. The prefabricated bar implant-retained overdenture showed low significant reduction in the bone height after 1 year, and a very highly significant reduction after 18 months. PMID- 20009605 TI - External hexagon and internal hexagon in straight and offset implant placement: strain gauge analysis. AB - PURPOSE: : The objective of this in vitro study was to quantify the strain development of external hexagon and internal hexagon implant-supported partial prostheses in straight and offset implant placement configurations. MATERIALS AND METHODS: : Three external hexagon and 3 internal hexagon implants were embedded in the center of each polyurethane block. Four strain gauges were bonded on the surface of polyurethane. The 20 superstructure occlusal screws were tightened onto the Microunit abutments with a torque of 10 Ncm using the manufacturer's manual of torque-controlling device. RESULTS: : There were statistic significant differences in hexagonal type (P = 0.0210 < 0.05). There were nostatistic significant differences in placement configuration (P = 0.7483 > 0.05). The results showed fundamental differences between both conditions. CONCLUSION: : Under the limited conditions of this study, the hexagon internal connection displayed higher values of microstrain than the hexagon external type implant placement. There was no evidence that there was an advantage of offset placement in reducing the strain around implant. There was no proof from this investigation that the straight placement generated higher microstrain than offset placement. PMID- 20009606 TI - Osteogenic potential of autogenous bone associated with bone marrow osteoblastic cells in bony defects: a histomorphometric study. AB - PURPOSE: Because of limitations of autogenous grafts, allografts, xenografts, alloplasts, and hydroxyapatite as graft materials, researchers have been using bone tissue engineering as a strategy for bone regeneration. The aim of this work was to study the effect of bone tissue engineering, associating bone marrow osteoblastic cells, and autogenous bone in defects created by dental extraction in rats. MATERIALS AND METHODS: Eighty male rats from 250 to 300 g were anesthetized, submitted to the extraction of the superior incisor, and divided in control group (C), implanted with osteoblastic cells (OC), autogenous bone (AB), and osteoblastic cells + autogenous bone (OC + AB). The animals were killed on 10th and 20th days after surgery and their maxilla were processed for obtaining fine semiserial sections (5 mum), and then stained with hematoxylin-eosin. Through image analysis system, bone volume in areas adjacent to the implants was estimated. RESULTS: The histometric results revealed that the association OC + AB produced significant increase (10%-15%) of bone in both experimental periods when compared with the control group (P < or = 0.01). CONCLUSIONS: Osteoblastic cells associated with autogenous bone accelerated the repair of bone defect, and the action of the osteoblastic cells was more effective until the 10th day and of the autogenous bone after this period. PMID- 20009632 TI - Qualitative analysis of immunization programs with most improved childhood vaccination coverage from 2001 to 2004. AB - State and urban immunization programs are responsible for the implementation of comprehensive programs to vaccinate populations within their geographic area. Given the variability in immunization coverage rates between geographic areas, the purpose of this two-phase study was to first identify the state and urban areas that achieved the highest increases in coverage, and then those with the highest sustained coverage, between two designated periods, and to interview key program staff members and their community counterparts to capture their perspectives on what factors may have contributed to increasing and sustaining high rates. In this article, we describe phase 1, in which we visited the seven sites that achieved the largest increases in coverage from 2001 to 2004. Results describe outcomes from the 71 semistructured key informant interviews with internal staff and external partners at the site's immunization programs. Interview transcripts were analyzed qualitatively, using a general inductive approach. Common challenges encountered among the seven sites included increasing reluctance among parents and overcoming barriers to accessing care. Common strategies to address these and other challenges included collecting and using data on immunization coverage, developing communication and education efforts, and continuously reaching out and collaborating with immunization partners. Lessons learned from these programs may help inform others who are working to improve childhood immunization delivery and coverage in their own programs. PMID- 20009607 TI - A prospective follow-up study of 44 mandibular immediately loaded implants using resonance frequency analysis: preliminary 1-year results. AB - PURPOSE: This clinical study aimed to evaluate initial, 4-months, and 1-year stability of immediately loaded dental implants inserted according to a protocol of lower rehabilitation with prefabricated bars. MATERIALS AND METHODS: The sample was composed of 11 edentulous patients. In each patient, 4 interforaminal implants were inserted. Immediately after implant installation, resonance frequency analysis (RFA) for each fixation was registered as well as after 4 months and 1 year with the prosthetic bar removed as it is a screwed system. RESULTS: The clinical implant survival rate was 100%. The RFA showed an increase in stability after 4 months from 64.09 +/- 6.48 to 64.31 +/- 4.96 and 1 year, 67.11 +/- 4.37. The analysis of variance showed a statistically significant result (P = 0.015) among implant stability quotient values for the different periods evaluated. Tukey test results showed statistically significant differences between 1-year results and the initial periods but there was no statistically significant difference between initial and 4-month results (P > 0.05). CONCLUSION: These preliminary 1-year results indicate that immediate loading of mandibular dental implants using the studied prefabricated bars protocol is a reliable treatment as it is in accordance with the results described in the literature for other similar techniques. PMID- 20009633 TI - A qualitative analysis of immunization programs with sustained high coverage, 2000-2005. AB - Despite record-high immunization coverage nationally, there is considerable variation across state and local immunization programs, which are responsible for the implementation of vaccine recommendations in their jurisdictions. The objectives of this study were to describe activities of state and local immunization programs that sustained high coverage levels across several years and to identify common themes and practical examples for sustaining childhood vaccination coverage rates that could be applied elsewhere. We conducted 95 semi structured key informant interviews with internal staff members and external partners at the 10 immunization programs with the highest sustained childhood immunization coverage from 2000 to 2005, as measured by the National Immunization Survey. Interview transcripts were analyzed qualitatively using a general inductive approach. Common themes across the 10 programs included maintaining a strong program infrastructure, using available data to drive planning and decision making, a commitment to building and sustaining relationships, and a focus on education and communication. Given the challenges of an increasingly complex immunization system, the lessons learned from these programs may help inform others who are working to improve childhood immunization delivery and coverage in their own programs. PMID- 20009634 TI - Quality improvement in public health: where are we now? PMID- 20009635 TI - Achieving a culture of quality improvement: the vision for public health in 2026. PMID- 20009636 TI - Defining quality improvement in public health. AB - Many industries commonly use quality improvement (QI) techniques to improve service delivery and process performance. Yet, there has been scarce application of these proven methods to public health settings and the public health field has not developed a set of shared principles or a common definition for quality improvement. This article discusses a definition of quality improvement in public health and describes a continuum of quality improvement applications for public health departments. Quality improvement is a distinct management process and set of tools and techniques that are coordinated to ensure that departments consistently meet the health needs of their communities. PMID- 20009637 TI - Opportunities to advance quality improvement in public health. PMID- 20009638 TI - Quality improvement and performance: CDC's strategies to strengthen public health. PMID- 20009639 TI - Supporting public health departments' quality improvement initiatives: lessons learned from the Public Health Foundation. PMID- 20009640 TI - The national public health performance standards: driving quality improvement in public health systems. AB - Since its inception in 1998, the Centers for Disease Control and Prevention's National Public Health Performance Standards Program (NPHPSP) has helped lay the groundwork for public health quality improvement (QI) activities at the state and local levels. This article describes how the NPHPSP has promoted QI through its instruments and guidance and how it has continually strengthened the focus on QI over the years. The NPHPSP Version 2 instruments and enhanced guidance have been designed to more strongly reinforce QI and catalyze the transition from assessment to action. Despite positive reports from some state and local users that emphasize the value the NPHPSP holds for those that do successfully move forward with improvement actions, 2005 evaluation results from the Association of State and Territorial Health Officials and the National Association of County and City Health Officials indicated challenges in transitioning the assessments results into performance improvement. More recent data are promising; a 2009 postassessment survey of early Version 2 respondents indicates that the majority (75% of all respondents) report action in one or more performance improvement steps. The NPHPSP has played an important role in fostering QI in many states and local jurisdictions. Furthermore, its experiences and lessons learned in supporting QI have helped to pave the way for other initiatives, such as the emerging national accreditation system for state and local health departments. PMID- 20009641 TI - Taking improvement action based on performance results: Washington State's experience. AB - Washington state used the Multistate Learning Collaborative grants to address improvement opportunities identified through the results of the Standards Performance Review cycles and county-specific local public health indicators. State and local health departments initiated statewide collaboratives to address the high priorities for system and individual health department improvements. Two individual agency quality improvement efforts are highlighted, one in a large local health department and the other in the State Department of Health, with emphasis on the results in establishing an agency quality improvement infrastructure and Rapid Cycle Improvement teams. PMID- 20009642 TI - State landscape in public health planning and quality improvement: results of the ASTHO survey. AB - Limited data exist on state public health agencies and their use of planning and quality improvement (QI) initiatives. Using the 2007 Association of State and Territorial Health Officials (ASTHO) State Public Health Survey, this article describes how state public health agencies perform tasks related to planning, performance management (PM), and QI. While 82 percent of respondents report having a QI process in place, only 9.8 percent have it fully implemented departmentwide. Seventy-six percent reported having a PM process in place, with 16 percent (n = 8) having it fully implemented departmentwide. A state health improvement plan was used by 80.4 percent of respondents, with 56.9 percent of respondents completing the plan more than 3 years ago. More than two-thirds (68.2%) of the respondents developed the plan by using results of their state health assessment. Analysis of state health department level planning, PM, and QI initiatives can inform states' efforts to ready themselves to meet the proposed national voluntary accreditation standards of the Public Health Accreditation Board. PMID- 20009643 TI - The quality improvement experience in a high-performing local health department: Los Angeles County. AB - Quality improvement in public health is a key element in the movement toward accreditation. Multiple national, state, and local initiatives are under way to define quality in a public health context and to develop tools and promising practices to support quality-improvement efforts in local health departments. Until recently, efforts to improve quality at the local level have largely focused on performance measurement to assess the relationship between inputs, outputs, and outcomes. The Los Angeles County Department of Public Health has developed its own unique approach to quality improvement. This approach includes focusing on three overlapping areas (professional practice, performance improvement, and public health science) that align closely with essential public health services 8 (competent worker), 9 (evaluation), and 10 (research). Broadening the focus of quality-improvement efforts to include these three areas (rather than performance improvement alone) provides additional opportunities to address key infrastructure issues that may affect the quality of services that are provided to the public and, thus, health outcomes. While the experience in Los Angeles County parallels other efforts, it includes unique elements that will be of use to public health professionals in other agencies. PMID- 20009644 TI - Quality improvement in local health departments: results of the NACCHO 2008 survey. AB - OBJECTIVES: To assess the current status of quality improvement (QI) within local health departments (LHDs) and examine the characteristics associated with such QI efforts. METHODS: A QI module was administered to a representative sample of 545 LHDs along with the core instrument in the 2008 NACCHO Profile survey of all LHDs nationally. Using the Profile survey data set, a quantitative approach was employed to determine the current status of QI within LHDs. Statistical analysis was performed to identify characteristics of LHDs associated with QI. The response rate to the QI module was 82 percent. RESULTS: Of the 448 LHDs that responded to the QI Module, 55 percent reported conducting formal QI efforts during the previous 2 years. Forty-four percent of these LHDs used a specific framework for QI, 56 percent used at least one of four commonly employed QI tools or techniques, and customer focus and satisfaction was the most frequently reported area (76%) of QI efforts. LHDs with large size of jurisdiction population and those with centralized governance were more likely to have engaged in quality or performance improvement, have managers who received formal QI training, and have provided QI training to staff. CONCLUSION: The 2008 NACCHO Profile QI module furnishes an excellent baseline for measuring progress of health department QI activities as accreditation and other related activities intensify. A clear definition of QI in public health that is understood by practitioners will greatly increase our ability to measure the adoption of QI by LHDs. Further research is necessary to identify and explore some of the predictors and possible barriers to increasing the application of QI by LHDs. PMID- 20009645 TI - Promoting quality improvement and achieving measurable change: the lead states initiative. AB - Along with the development of a national voluntary accreditation program for public health departments that holds quality improvement as its core goal, the application of quality improvement in public health has been gaining momentum. The 16 states participating in the Multi-State Learning Collaborative: Lead States in Public Health Quality Improvement (MLC) represent best practices in these activities. The MLC brings together partnerships in 16 US states to prepare for accreditation and implement quality-improvement practices. The grantee states are managing quality-improvement teams of local and state health department representatives and other partners. These teams, called mini-collaboratives, are working collectively to implement quality-improvement techniques to make measurable change on identified public health issues, or target areas. The work of the MLC seems to show that state and local-health departments and their key partners have the leadership, will and interest to apply quality improvement tools, and methods to solving public health problems and to raising the standard of public health practice. This article describes the history, current status, and lessons learned from the work of the MLC. PMID- 20009646 TI - Evaluating the Multi-State Learning Collaborative. AB - The third phase of the Multi-State Learning Collaborative (MLC) is a 3-year Robert Wood Johnson Foundation initiative designed to build quality improvement capacity within state and local health departments. This commentary provides a brief overview of the MLC, a methodological description of its evaluation, and preliminary findings. Major evaluation data sources include quarterly reports, surveys, case studies, and key informant interviews. Preliminary survey data and quarterly reports from year 1 provide some early evidence that states are progressing toward MLC goals. Approximately 27 percent of local health departments in the 16 participant states report that they have begun to prepare for national accreditation and approximately 39 percent report implementation of at least one quality improvement project within the past 12 months. Ongoing data collection efforts are underway to more fully address the evaluation questions. PMID- 20009647 TI - Driving quality improvement in local public health practice. AB - OBJECTIVES: To understand what tools, resources, and assistance are needed for local health departments (LHDs) to successfully engage in quality improvement (QI) and to generate examples of successful QI efforts. METHODS: With funding from the Centers for Disease Control and Prevention and the Robert Wood Johnson Foundation, the National Association of County & City Health Officials supported 66 LHD demonstration sites between 2007 and 2009. The sites measured themselves against national standards and addressed priority areas for improvement through the application of QI techniques. We used on-line surveys, interviews, and informal collection of participant feedback to determine the usefulness of resources that were provided for QI efforts. RESULTS: Participating LHDs lack a common understanding of formal QI. Several existing QI resources specifically geared to public health are very useful, and in-person assistance is highly valued. DISCUSSION: The value of Web-based sessions is uncertain, and state and national meetings could provide accessible forums for in-person training. Dedicated time to training and implementation, coupled with widespread sharing of best practices and success stories, could enhance the uptake of QI efforts in LHDs. Additional studies regarding sustainability are needed to understand how to institutionalize QI. PMID- 20009648 TI - Realizing transformational change through quality improvement in public health. AB - OBJECTIVE: This article discusses the specific components necessary to achieve transformational change within public health departments as a means for creating sustained performance improvement and better outcomes in the health of the community. DESIGN: This article provides a review of transformation change concepts and application to public health departments. RESULTS: Transformational change for public health departments must be intentionally designed to achieve high performance. While all improvement requires change, not all change results in improvement. CONCLUSION: The successful transformational change effort always occurs from the top-down, while the process improvement occurs from the bottom up. Transformational change is possible in public health departments when small incremental improvements are linked with large-scale management changes to continually improve public health performance resulting in better population outcomes. PMID- 20009649 TI - Quality improvement and accreditation: what might it look like? AB - Following the 2006 release of the Exploring Accreditation Project report on the desirability and feasibility of establishing a national voluntary public health accreditation program, the Public Health Accreditation Board was established. Since that time, standards, measures, and an assessment process have been developed. Beta testing of the full accreditation cycle is underway. The Public Health Accreditation Board accreditation program relies heavily on quality improvement as its cornerstone. In this commentary, we describe how accreditation and quality improvement can operate together synergistically to strengthen public health practice and improve outcomes at the community level. PMID- 20009650 TI - Achieving a culture of quality improvement. AB - The National Association of County and City Health Officials (NACCHO) is the national organization representing local health departments. It supports efforts that protect and improve the health of all people and all communities by promoting national policy, developing resources and programs, seeking health equity, and supporting effective local public health practice and systems. PMID- 20009651 TI - Successful laparoscopic repair of acute intrapericardial diaphragmatic hernia secondary to penetrating trauma. PMID- 20009652 TI - Islet autotransplantation after distal pancreatectomy for pancreatic trauma. PMID- 20009653 TI - Early laparoscopic approach for complicated blunt hepatic injury in an adolescent. PMID- 20009654 TI - Isolated injury of the posterior interosseous nerve complicating a deep laceration of the proximal forearm dorsolateral muscles: a case report. PMID- 20009655 TI - Abdominal shotgun wound with pellet embolization leading to bilateral lower limb amputation: case report and review of the literature of missile emboli over the past 10 years. PMID- 20009656 TI - Severe fetal skull fracture and death subsequent to a motor vehicle crash with frontal airbag deployment. AB - Frontal airbag deployment may cause severe fetal injury and loss without maternal injury. PMID- 20009657 TI - A gift: Presidential Address at the 2009 Western Trauma Association for the surgery of trauma. AB - This Western Trauma Association Presidential Address is about my mentor program. It outlines some background material on mentorship and the results of an alumni mentee survey. PMID- 20009658 TI - Western Trauma Association critical decisions in trauma: nonoperative management of adult blunt hepatic trauma. PMID- 20009659 TI - Western Trauma Association critical decisions in trauma: screening for and treatment of blunt cerebrovascular injuries. PMID- 20009660 TI - Pulseless electrical activity, focused abdominal sonography for trauma, and cardiac contractile activity as predictors of survival after trauma. AB - BACKGROUND: Pulseless electrical activity (PEA) secondary to both blunt and penetrating trauma is associated with minimal survival. The pericardial view of the focused abdominal sonography for trauma (p-FAST) can differentiate between patients with and without organized cardiac activity and may assist in the decision to terminate ongoing resuscitation. METHODS: A retrospective review was performed for all patients presenting to a level I trauma center from January 2006 through January/2009 who had PEA on arrival or developed PEA in the emergency department. Additional data abstracted included outcome, the p-FAST findings, and mechanism of injury. Recorded FAST examinations were reviewed by a blinded ultrasound trained physician. RESULTS: During the study period 25 patients presented with PEA and three developed PEA during initial resuscitation. Contractile cardiac activity was present in nine patients with PEA on presentation and immediately after deterioration to PEA in the three patients developing PEA. Four patients had a penetrating mechanism and 24 were blunt. Two pericardial effusions were present on examination, both after blunt trauma. Three patients survived beyond the emergency department (89% early mortality). The survivors had presented in PEA with organized cardiac contractile activity on ultrasound and had tension pneumothorax, tension hemothorax, and hypovolemia treated. Two patients died in the operating room of uncontrolled hemorrhage and one patient died 6 days after admission because of closed head injury. CONCLUSIONS: The presence of PEA at any time during initial resuscitation is a grave prognostic indicator. p-FAST is a useful test to identify contractile cardiac activity. p-FAST may identify those patients with potential for survival. PMID- 20009661 TI - A major metropolitan "field amputation" team: a call to arms ... and legs. AB - BACKGROUND: As early as 1979, suggestions were made to establish amputation teams and protocols in major metropolitan areas. It was recognized that preplanning on such calls would be valuable to carrying out rescues of that nature. Since then, questionnaires and collegial conversations reveal the existence of such teams remains the exception in our nation's cities. METHODS: Our team was formed in 1984 after an emergency medical service request for a surgeon to perform an amputation on a person who had become entrapped with both arms in an industrial candy press was made. In its current form, the team consists of an attending trauma surgeon, a resident surgeon, a registered nurse, and a pilot, all hospital based. Equipment is limited to medications for sedation and pain control, two units of uncross-matched blood, and a prebundled duffle bag of bandages, a scalpel, various saws, and hemostats. Transportation to the scene is provided by the helicopter based at our level II trauma center. RESULTS: Since its inception, the team has been activated three to four times per year, resulting in nine amputation rescues. Three of these cases, presented here, are from an unusually busy 5 weeks during the spring of 2008. The first case involves a tree shredding device, the second, an industrial auger, and the third, a forklift and a steel toed boot. In these cases, the utilization of the amputation team resulted in successful patient rescues and outcomes. CONCLUSION: A field amputation team can be an integral part of any emergency medical service system, filling an infrequently used but helpful adjunct to emergency care. PMID- 20009662 TI - Burns, inflammation, and intestinal injury: protective effects of an anti inflammatory resuscitation strategy. AB - BACKGROUND: Intestinal barrier breakdown after severe burn can lead to intestinal inflammation, which may act as the source of the systemic inflammatory response. In vitro intestinal cell studies have shown that mitogen-activated protein kinase (MAPK) signaling is an important modulator of intestinal inflammation. We have previously observed that pentoxifylline (PTX) attenuates burn-induced intestinal permeability and tight junction breakdown. We hypothesized that PTX would limit intestinal barrier breakdown and attenuate inflammatory signaling via the MAPK pathway. METHODS: Male balb/c mice underwent 30% total body surface area full thickness steam burn. Immediately after burn, animals received an intraperitoneal injection of PTX (12.5 mg/kg) in normal saline or normal saline alone. In vivo intestinal permeability to 4 kDa fluorescein isothiocyanate-dextran was measured. Intestinal extracts were obtained to measure interleukin-6 by enzyme-linked immunosorbent assay, and phosphorylated p38 MAPK, p38 MAPK, phosphorylated extracellular signal-related kinase (1/2) (ERK (1/2)), and ERK (1/2) by immunoblotting. Acute lung injury was assessed by histology at 24 hours after burn. RESULTS: Administration of PTX immediately after injury attenuated burn induced intestinal permeability. PTX also decreased the burn-induced phosphorylation of p38 MAPK and decreased phosphorylation of ERK (1/2) at 2 hours and 24 hours after injury. Animals given PTX had decreased intestinal interleukin 6 levels. A single dose of PTX also decreased histologic lung injury at 24 hours after burn. CONCLUSION: PTX attenuates burn-induced intestinal permeability and subsequent intestinal inflammation. Use of PTX after burn was also associated with decreased acute lung injury. Because of its compelling anti-inflammatory effects, PTX may be an ideal candidate for use as an immunomodulatory adjunct to resuscitation fluid. PMID- 20009663 TI - Where do we go from here? Interim analysis to forge ahead in violence prevention. AB - BACKGROUND: The severity and disparity of interpersonal violent injury is staggering. Fifty-three per 100,000 African Americans (AA) die of homicide yearly, 20 per 100,000 in Latinos, whereas the rate is 3 per 100,000 in Caucasians. With the ultimate goal of reducing injury recidivism, which now stands at 35% to 50%, we have designed and implemented a hospital-based, case managed violence prevention program uniquely applicable to trauma centers. The Wraparound Project (WP) seizes the "teachable moment" after injury to implement culturally competent case management (CM) and shepherd clients through risk reduction resources with city and community partners. The purpose of this study was to perform a detailed intermediate evaluation of this multi-modal violence prevention program. We hypothesized that this evaluation would demonstrate feasibility and early programmatic efficacy. We looked to identify areas of programmatic weakness that, if corrected, could strengthen the project and enhance its effectiveness. METHODS: We performed intermediate evaluation on the 18-month-old program. We selected the Centers for Disease Control and Prevention recommended instrument used for unintentional injury prevention programs and applied it to the WP. The four sequential stages in this methodology are formative, process, impact, and outcome. To test feasibility of WP, we used process evaluation. To evaluate intermediate goals of risk reduction and early efficacy, we used impact evaluation. RESULTS: Four hundred thirty-five people met screening criteria. The two case managers were able to make contact and screen 73% of gun shot victims, and 57% of stab wound victims. Of those not seen, 48% were in the hospital for 6 h/wk with the client. Forty-one percent of the time, they spent 3 hours to 6 hours. Seventeen of 18 people who required >6 hours had two to three needs. Attrition rate is only 4%. The table demonstrates percent success thus far in providing risk reduction resources. CONCLUSIONS: WP case managers served high-risk clients by developing trust, credibility, and a risk reduction plan. Cultural competency has been vital. Six of seven major needs were successfully addressed at least 50% of the time. The value of reporting these results has led WP to gain credibility with municipal stakeholders, who have now agreed to fund a third CM position. Intermediate evaluation provided a framework in our effort to achieve the ultimate goal of reducing recidivism through culturally competent CM and risk factor modification. PMID- 20009664 TI - Protein C depletion early after trauma increases the risk of ventilator associated pneumonia. AB - INTRODUCTION: Mechanically ventilated trauma patients have a high risk for the development of ventilator-associated pneumonia (VAP). We have recently reported that reduced plasma protein C (PC) levels early after trauma/shock are associated with coagulopathy and mortality. Furthermore, trauma patients with tissue injury and shock are at higher risk for the development of VAP. OBJECTIVE: We hypothesized that low PC levels early after trauma are associated with an increased susceptibility to VAP in trauma patients. METHODS: Fifty-nine acutely injured, intubated trauma patients were admitted to the critical care unit. Serial blood samples were drawn and coagulation factors were measured. VAP was diagnosed by presence of bacteria on bronchial alveolar lavage specimen, bilateral infiltrates on chest roentgenogram, and fever or elevated white blood cell count. RESULTS: There were no differences in demographic or injury characteristics between patients who developed VAP and those who did not. As expected, patients who developed VAP had more ventilator days, hospital days, intensive care unit days, and greater mortality (all p < 0.05). Patients in both groups had lower mean PC levels at 6 hours compared with baseline. Noninfected patients' PC subsequently returned to near baseline levels, whereas those patients who eventually acquired VAP had significantly lower PC levels at both 12 and 24 hours (12 hours: 79 vs. 96%, p = 0.05; 24 hours: 75 vs. 97% p = 0.02). Soluble endothelial PC receptor (sEPCR) levels were also lower at 24 hours (82 vs. 99% in the noninfected group, p = 0.04). DISCUSSION: The activation of PC pathway early after trauma may protect the vascular endothelium by both its anticoagulant and cytoprotective effects. However, trauma patients who later developed VAP have significantly lower plasma levels of PC within 24 hours after injury, suggesting a possible consumption of this vitamin K-dependent protein and an inhibition of its activation by inflammatory mediators. EPCR is involved in the activation of PC and is also a mediator of its cytoprotective effects. CONCLUSION: Critically ill trauma patients have an early activation of the PC pathway, associated with a rapid decrease in the plasma levels of this protein and increase in EPCR. Plasma levels of PC return to normal levels within 24 hours in most patients. However, patients who go on to acquire VAP have persistently low plasma levels of PC in the immediate period after trauma. Whether PC could play a mechanistic role in the host response against nosocomial lung infection warrants further study. PMID- 20009666 TI - Trauma attenuating backing improves protection against behind armor blunt trauma. AB - BACKGROUND: Body armor is used by military personnel, police officers, and security guards to protect them from fatal gunshot injuries to the thorax. The protection against high-velocity weapons may, however, be insufficient. Complementary trauma attenuating backings (TAB) have been suggested to prevent morbidity and mortality in high-velocity weapon trauma. METHODS: Twenty-four Swedish landrace pigs, protected by a ceramid/aramid body armor without (n = 12) or with TAB (n = 12) were shot with a standard 7.62-mm assault rifle. Morphologic injuries, cardiorespiratory, and electroencephalogram changes as well as physical parameters were registered. RESULTS: The bullet impact caused a reproducible behind armor blunt trauma (BABT) in both the groups. The TAB significantly decreased size of the lung contusion and prevented hemoptysis. The postimpact apnea, desaturation, hypotension, and rise in pulmonary artery pressure were significantly attenuated in the TAB group. Moreover, TAB reduced transient peak pressures in thorax by 91%. CONCLUSIONS: Our results indicate that ordinary body armor should be complemented by a TAB to prevent thoracic injuries when the threat is high-velocity weapons. PMID- 20009665 TI - Predictive modeling and inflammatory biomarkers in rats with lung contusion and gastric aspiration. AB - BACKGROUND: This study uses statistical predictive modeling and hierarchical cluster analyses to examine inflammatory mediators and cells in bronchoalveolar lavage (BAL) as putative biomarkers in rats with blunt trauma lung contusion (LC), gastric aspiration (combined acid and small gastric food particles, CASP), or a combination of the two. METHODS: Specific parameters assessed in the innate pulmonary inflammatory response were leukocytes, macrophages, and polymorphonuclear neutrophils (PMNs) in BAL; whole lung myeloperoxidase activity; and a series of cytokines or chemokines present in BAL at 5 or 24 hours after injury: tumor necrosis factor-alpha, interleukin (IL)-1beta, IL-6, interferon gamma, IL-10, macrophage inflammatory protein-2, cytokine-induced neutrophil chemoattractant-1, and monocyte chemoattractant protein-1. RESULTS: Rats with LC, CASP, LC + CASP all had severe lung injury compared with uninjured controls based on decreased arterial oxygenation or increased BAL albumin at 5 or 24 hours postinsult. However, the injury groups had distinct overall patterns of inflammation that allowed them to be discriminated accurately by hierarchical cluster analysis (29 of 30 and 35 of 37 rats were correctly classified in hierarchical clusters at 5 and 24 hours, respectively). Moreover, predictive analyses based on an extension of standard receiver-operator characteristic methodology discriminated individual animals and groups with similar high accuracy based on a maximum of two inflammatory parameters per group (29 of 30 and 36 of 37 rats were correctly classified at 5 hours and 24 hours, respectively). CONCLUSIONS: These results support the possibility that inflammatory biomarker profiles could be developed in the future to improve the diagnosis and management of trauma patients with unwitnessed (occult) gastric aspiration who have an increased risk of clinical acute lung injury or the acute respiratory distress syndrome. PMID- 20009667 TI - Comparison of the plasma volume-expanding effects of 6% dextran 70, 5% albumin, and 6% HES 130/0.4 after hemorrhage in the guinea pig. AB - BACKGROUND: We still lack comparing data of the plasma volume (PV)-expanding effect of the most commonly used colloids including dextran 70. This study compares the PV-expanding effects of 6% dextran 70, 5% albumin, and 6% hydroxyethylstarch (HES) 130/0.4 after a standardized hemorrhage. METHODS: The prospective and randomized study on 33 anesthetized adult male guinea pigs involved three groups (n = 11 each); the dextran group, the albumin group, and the HES group. The left carotis artery was cannulated for blood pressure measurements and blood samples, and the right jugular vein was cannulated for infusions. After hemorrhage of 20 mL/kg for 8 minutes, the animals were transfused with 20 mL/kg of the colloid for 10 minutes. PV was determined with a I-albumin tracer dilution technique at baseline and 3 hours after the colloid infusion. The PV just after hemorrhage was calculated as the baseline value minus bled PV. Blood gases were measured at baseline, after hemorrhage, just after the colloid infusion and at the end of the experiment. RESULTS: The increase in PV 3 hours after the colloid infusion, including the 20 mL infused, was 36.3 mL/kg +/- 2.3 mL/kg in the dextran group, 26.4 mL/kg +/- 4.7 mL/kg in the albumin group, and 17.6 mL/kg +/- 3.5 mL/kg in the HES group. At the end of the experiment, hematocrit was lower in the dextran group than in the albumin and the HES groups. Urine production was higher in the HES group than in the dextran and the albumin groups. CONCLUSION: After hemorrhage, the PV-expanding capacity of 6% dextran 70 was better than that of 5% albumin, which was in turn better than that of HES 130/0.4 given in equal volumes. PMID- 20009668 TI - Fluid replacement with hypertonic or isotonic solutions guided by mixed venous oxygen saturation in experimental hypodynamic sepsis. AB - BACKGROUND: Splanchnic perfusion is prone to early injury and persists despite normalization of global hemodynamic variables in sepsis. Volume replacement guided by oxygen derived variables has been recommended in the management of septic patients. Our hypothesis was that a hypertonic isoncotic solution would improve the benefits of crystalloids replacement guided by mixed venous oxygen saturation. METHODS: Seventeen anesthetized and mechanically ventilated mongrel dogs received an intravenous infusion of live E. coli in 30 minutes. They were then randomized into three groups: control group (n = 3) bacterial infusion without treatment; normal saline (n = 7), initial fluid replacement with 32 mL/kg of normal saline during 20 minutes; hypertonic solution (n = 7), initial fluid replacement with 4 mL/kg of hypertonic solution during 5 minutes. After 30 and 60 minutes, additional boluses of normal saline were administered when mixed venous oxygen saturation remained below 70%. Mean arterial pressure, cardiac output; regional blood flows, systemic and regional oxygen-derived variables, and lactate levels were assessed. Animals were observed for 90 minutes and then killed. Hystopathological analysis including apoptosis detection using terminal deoxynucleotidil transferase mediated dUTP-biotin nick end labeling was performed. RESULTS: A hypodynamic septic shock was observed after bacterial infusion. Both the fluid-treated groups presented similar transient benefits in systemic and regional variables. A greater degree of gut epithelial cells apoptosis was observed in normal saline-treated animals. CONCLUSIONS: Although normalization of mixed venous oxygen saturation was not associated with restoration of markers of splanchnic or other systemic perfusion variables, the initial fluid savings with hypertonic saline and its latter effect on gut apoptosis may be of interest in sepsis management. PMID- 20009669 TI - Effects of different resuscitation fluids on acute lung injury in a rat model of uncontrolled hemorrhagic shock and infection. AB - BACKGROUND: To investigate the effects of different resuscitation fluids on acute lung injury in a rat model of uncontrolled hemorrhagic shock (HS) followed by mimicked infection. METHODS: Sixty Sprague-Dawley rats were randomly assigned to one of the five groups (n = 12 per group) to receive the following treatments: (1) control group (group C), surgery, no hemorrhage, and no resuscitation; (2) no fluid resuscitation group; (3) lactated Ringer's solution group; (4) 7.5% hypertonic saline (HTS) group; and (5) hydroxyethyl starch group (group HES). All experimental groups were subjected to three phases: phase I entailed massive hemorrhage with a mean arterial pressure of 35 mm Hg to 40 mm Hg for 60 minutes by tail amputation and followed by mimicked infection by intratracheal administration of lipopolysaccharide 2 mg/kg. The animals in each group were then partially resuscitated with the fluid assigned to the group. Phase II of 60 minutes commenced at tail ligation, involved hemostasis, and return of all the blood initially shed. Phase III was an observation phase with no any further treatment and lasted for 3.5 hours. The survival rate at the end of the phase III was recorded. After phase III, arterial blood gases were recorded. The wet to dry lung weight ratio, pulmonary microvascular permeability, the expression of transforming growth factor (TGF)-beta1, and Smad2 were determined. The lung histology was also assessed. RESULTS: HES and HTS solutions were more effective than no fluid resuscitation and lactated Ringer's solution in reducing the detrimental effects of HS and infection on the lungs, as seen by the significantly lower pulmonary microvascular permeability and wet to dry lung weight ratio, the improved arterial blood gases and lower levels of TGF-beta1and Smad2 expression in lung tissues. These beneficial effects were most pronounced in the group HES. CONCLUSIONS: This study demonstrated that resuscitation with HTS and especially with HES could reduce lung tissue damage and pulmonary edema after severe uncontrolled HS. The TGF-beta1/Smad2 signaling pathway might play a key role in regulation of pulmonary permeability and formation of pulmonary edema in a rat model of uncontrolled HS and infection. PMID- 20009670 TI - Influence of extravascular lung water determination in fluid and vasoactive therapy. AB - INTRODUCTION: Preload parameters in postresuscitation phase are not sufficiently sensitive to guide fluid therapy in critically ill patients. We analyzed modifications in the fluid therapy and vasoactive drugs of critically ill patients that were produced by inclusion of extravascular lung water (EVLW) data in the treatment protocol and evaluated the short-term response. METHODS: This observational and prospective study included consecutive patients with hypotension or hypoxemia, comparing the therapeutic plan for fluid and vasoactive drug treatment between before and after knowing the EVLW value. RESULTS: We studied 42 patients. After knowing the EVLW, 52.4% (n = 22) of initial therapeutic plans were changed, modifying fluid therapy in all of these cases and vasoactive therapy in 22% of them. EVLW value was 13.91 +/- 5.62 in patients with change of therapeutic plan versus 10 +/- 4.52 in those with no change (p < 0.05). No differences were found in preload parameters as a function of change/no change. The most frequent decision change (n = 13) was to fluid reduction plus diuretic administration, and patients with this modification had significantly (p < 0.05) higher EVLW values compared with the remaining patients with a change in fluid therapy. Out of the 22 patients with a modified therapeutic decision, the therapy proved effective in 18 patients CONCLUSION: Quantification of EVLW in patients who can be considered euvolemic induces important modifications in fluid and vasoactive therapy. These changes generally resulted in a lower volume loading and a positive outcome for the patient. PMID- 20009671 TI - A comparison between pulse pressure variation and right end diastolic volume index as guides to resuscitation in a model of hemorrhagic shock in pigs. AB - BACKGROUND: Different hemodynamic parameters including static indicators of cardiac preload as right ventricular end-diastolic volume index (RVEDVI) and dynamic parameters as pulse pressure variation (PPV) have been used in the decision-making process regarding volume expansion in critically ill patients. The objective of this study was to compare fluid resuscitation guided by either PPV or RVEDVI after experimentally induced hemorrhagic shock. METHODS: Twenty-six anesthetized and mechanically ventilated pigs were allocated into control (group I), PPV (group II), or RVEDVI (group III) group. Hemorrhagic shock was induced by blood withdrawal to target mean arterial pressure of 40 mm Hg, maintained for 60 minutes. Parameters were measured at baseline, time of shock, 60 minutes after shock, immediately after resuscitation with hydroxyethyl starch 6% (130/0.4), 1 hour and 2 hours thereafter. The endpoint of fluid resuscitation was determined as the baseline values of PPV and RVEDVI. Statistical analysis of data was based on analysis of variance for repeated measures followed by the Bonferroni test (p < 0.05). RESULTS: Volume and time to resuscitation were higher in group III than in group II (group III = 1,305 +/- 331 mL and group II = 965 +/- 245 mL, p < 0.05; and group III = 24.8 +/- 4.7 minutes and group II = 8.8 +/- 1.3 minutes, p < 0.05, respectively). All static and dynamic parameters and biomarkers of tissue oxygenation were affected by hemorrhagic shock and nearly all parameters were restored after resuscitation in both groups. CONCLUSION: In the proposed model of hemorrhagic shock, resuscitation to the established endpoints was achieved within a smaller amount of time and with less volume when guided by PPV than when guided by pulmonary artery catheter-derived RVEDVI. PMID- 20009672 TI - Mechanical ventilation and fluid retention in burn patients. AB - BACKGROUND: Burn patients with inhalation injury (INHI) require more fluid resuscitation than patients without INHI. However, the relation between INHI and fluid resuscitation may be confounded by a ventilation-induced increase in fluid retention. We therefore evaluated whether INHI was independently of continuous positive pressure ventilation (CPPV) associated with increased fluid retention. METHODS: One hundred eighty-six patients with burns of >20% of total body surface area admitted to the Beverwijk Burns Center (1995-2006) were retrospectively studied. Cumulative fluid balance, defined as the total volume of fluids administered from the time of admission minus the total volume of fluids collected from each patient, was calculated at the end of days 3 (FB3) and 7 (FB7) postburn. The population was divided into three groups: (1) INHI-CPPV- (no INHI, no ventilation; n = 75); (2) INHI-CPPV+ (no INHI with ventilation; n = 62); and (3) INHI+CPPV+ (INHI with ventilation; n = 49). Analyses were corrected for differences in age, weight, and % total body surface area. RESULTS: Patients who were mechanically ventilated were older and had more extensive burns than those who were not ventilated. Baseline characteristics of patients without INHI who were treated by CPPV were similar to patients with INHI, also treated by CPPV. FB3 was significantly higher in patients without INHI who were ventilated compared with nonventilated patients (13.4 +/- 5.8 L vs. 23.1 +/- 10.6 L for INHI CPPV- and INHI-CPPV+ respectively, p = 0.001). However, fluid balance was not additionally affected by the presence of INHI. The difference in fluid retention between nonventilated and ventilated patients was also seen on day 7 (22.1 +/- 9.4 L vs. 34.2 +/- 15.9 L for INHI-CPPV- and INHI-CPPV+, respectively, p = 0.001). CONCLUSION: These results suggest that increased fluid retention, which is conventionally associated with INHI, is due to the effects of ventilation and not to the effects of INHI itself. This warrants a closer evaluation of patients who are ventilated in the absence of INHI, with a view to early extubation. PMID- 20009673 TI - Acute respiratory distress syndrome in nontrauma surgical patients: a 6-year study. AB - BACKGROUND: Acute respiratory distress syndrome (ARDS) has been shown to increase morbidity but not mortality in trauma patients; however, little is known about the effects of ARDS in nontrauma surgical patients. The purpose of this study is to evaluate the risk factors for and outcomes of ARDS in nontrauma surgical patients. STUDY: A prospective observational study was performed in the surgical intensive care unit (ICU) of an academic tertiary care center. From 2000 to 2005, all nontrauma surgical admissions to the surgical ICU were evaluated daily for ARDS based on predefined diagnostic criteria. Logistic regression analysis identified independent predictors for ARDS and ICU mortality. RESULTS: Of 2,046 patient identified, 125 (6.1%) met criteria for ARDS. The incidence of ARDS declined annually from 12.2% to 2.1% during the study period (p < 0.001). ARDS patients were significantly older (55.4 years vs. 51.8 years, p = 0.014) and more likely to be obese (32% vs. 22%, p = 0.007) than the non-ARDS population. Independent predictors of ARDS included use of pressors (relative risk, RR = 3.30), sepsis (RR = 1.72), and body mass index >or=30 kg/m (RR = 1.57). Independent predictors of ICU mortality included ARDS (RR = 6.88), pressors (RR = 2.85), positive fluid balance (RR = 2.27), Acute Physiology and Chronic Health Evaluation II (RR = 1.04), and age (RR = 1.02). CONCLUSIONS: Unlike trauma patients, ARDS was an independent predictor of ICU mortality in nontrauma surgical patients, independent of age and disease severity. Nontrauma surgical patients who developed ARDS were older, sicker, and had a longer ICU stay. Independent predictors of ARDS included use of pressors, sepsis, and obesity. PMID- 20009674 TI - Emergency department thoracotomy for penetrating injuries of the heart and great vessels: an appraisal of 283 consecutive cases from two urban trauma centers. AB - BACKGROUND: Historically, patients with penetrating cardiac injuries have enjoyed the best survival after emergency department thoracotomy (EDT), but further examination of these series reveals a preponderance of cardiac stab wound (SW) survivors with only sporadic cardiac gunshot wound (GSW) survivors. Our primary study objective was to determine which patients requiring EDT for penetrating cardiac or great vessel (CGV) injury are salvageable. METHODS: All patients who underwent EDT for penetrating CGV injuries in two urban, level I trauma centers during 2000 to 2007 were retrospectively reviewed. Demographics, injury (mechanism, anatomic injury), prehospital care, and physiology (signs of life [SOL], vital signs, and cardiac rhythm) were analyzed with respect to hospital survival. RESULTS: The study population (n = 283) comprised young (mean age, 27.1 years +/- 10.1 years) men (96.1%) injured by gunshot (GSW, 88.3%) or SWs (11.7%). Patients were compared by injury mechanism and number of CGV wounds with respect to survival (SW, 24.2%; GSW, 2.8%; p < 0.001; single, 9.5%; multiple, 1.4%; p = 0.003). Three predictors-injury mechanism, ED SOL, and number of CGV wounds-were then analyzed alone and in combination with respect to hospital survival. Only one patient (0.8%) with multiple CGV GSW survived EDT. CONCLUSION: When the cumulative impact of penetrating injury mechanism, ED SOL, and number of CGV wounds was analyzed together, we established that those sustaining multiple CGV GSWs (regardless of ED SOL) were nearly unsalvageable. These results indicate that when multiple CGV GSWs are encountered after EDT, further resuscitative efforts may be terminated without limiting the opportunity for survival. PMID- 20009675 TI - Blunt cardiac trauma: lessons learned from the medical examiner. AB - OBJECTIVE: The objective of this study was to analyze autopsy findings after blunt traumatic deaths to identify the incidence of cardiac injuries and describe the patterns of associated injuries. METHODS: All autopsies performed by the Los Angeles County Forensic Medicine Division for blunt traumatic deaths in 2005 were retrospectively reviewed. Only cases that underwent a full autopsy including internal examination were included in the analysis. The study population was divided into two groups according to the presence or absence of a cardiac injury and compared for differences in baseline characteristics and types of associated injuries. RESULTS: Of the 881 fatal victims of blunt trauma received by the Los Angeles County Forensic Medicine Division, 304 (35%) underwent a full autopsy with internal examination and were included in the analysis. The mean age was 43 years +/- 21 years, patients were more often men (71%) and were intoxicated in 39% of the cases. The most common mechanism was motor vehicle collision (50%), followed by pedestrian struck by auto (37%), and 32% had a cardiac injury. Death at the scene was significantly more common in patients with a cardiac injury (78% vs. 65%, p = 0.02). The right chambers were the most frequently injured (30%, right atrium; 27%, right ventricle). Among the 96 patients with cardiac injuries, 64% had transmural rupture. Multiple chambers were ruptured in 26%, the right atrium in 25%, and the right ventricle in 20% of these patients. Patients with cardiac injuries were significantly more likely to have other associated injuries: thoracic aorta (47% vs. 27%, p = 0.001), hemothorax (81% vs. 59%, p < 0.001), rib fractures (91% vs. 71%, p < 0.001), sternum fracture (32% vs. 13%, p < 0.001), and intra-abdominal injury (77% vs. 48%, p < 0.001) compared with patients without cardiac injury. Of the 96 patients with a cardiac injury, 78% died at the scene of the crash and 22% died en route or at the hospital. CONCLUSION: Cardiac injury is a common autopsy finding after blunt traumatic fatalities, with the majority of deaths occurring at the scene. Patients with cardiac injuries are at significantly increased risk for associated thoracic and intra-abdominal injuries. PMID- 20009676 TI - Firearm-related hunting accidents in Denmark. AB - BACKGROUND: Hunting-related accidents occur worldwide during the hunting season. The "true incidence" of hunting-related accidents in Denmark is not illuminated in that no compulsory systematic registration procedure is available. The possible impact of alcohol consumption on the risk of being involved in a hunting accident is not yet known; however, the subject frequently is debated throughout the world. METHODS: From a cross-sectional survey including 1,797 Danish hunters over a 5-year period (1997-2002), we found 71 hunting-related accidents where the hunter was either witness to or directly involved in the event. In 32 cases, the hunter was directly involved in the event. We estimated the incidence and risk of being involved in a hunting-related accident. We performed logistic regression analysis to reveal the possible impact of selected effect variables. Level of significance was 95%. RESULTS: The yearly incidence of being involved in an accident is 4.0 per 1,000 Danish hunters. The projected cumulated risk of a 21 year-old hunter to be involved in at least one hunting-related accident during a period of 20 years and 40 years is 7% and 14%, respectively. We found a significantly increased risk of being directly involved in or witness to a hunting-related accident with increased alcohol intake. CONCLUSIONS: The incidence of hunting-related accidents in Denmark seems considerably higher than reported. Compulsory and systematic registration of all hunting-related accidents involving firearms should be introduced in Denmark. Legislation rules regarding alcohol intake during hunting should be considered by the authorities. PMID- 20009677 TI - Admission base deficit as a long-term prognostic factor in severe pediatric trauma patients. AB - BACKGROUND: Base deficit (BD) is a prognostic tool that correlates with trauma scores and mortality in adult trauma patients. Retrospective studies have shown that admission BD more than 8 mmol/L is associated with an increased risk of mortality. This is the first prospective European study aimed at evaluating the prognostic value of admission BD in traumatized children. METHODS: One hundred severely traumatized children were included if an arterial BD had been calculated on arrival in the trauma room of a university hospital. Epidemiologic, medical, and biological data (including admission BD and lactates concentration) were recorded and compared using a univariate analysis. The primary endpoint was in hospital mortality. Secondary endpoints were outcome on discharge and at 6 months. Cutoff values for BD or lactates regarding outcomes were determined using receiver operating characteristic curves if these data had been isolated on multivariate analysis (p < 0.05). RESULTS: Sixty-eight boys and 32 girls, aged 6.7 years, were enrolled from March 2003 to December 2005, mainly after road traffic accidents. Twenty-two died at the hospital, 34 children and 51 children were classified as having a good outcome on hospital discharge and 6 months later, respectively. After the multivariate procedure and receiver operating characteristic curve analysis, admission lactates more than 2.94 mmol/L and admission BD more than 5 mEq/L were independent risk factors for mortality (odds ratio 2.4 [95% confidence interval 1.3-4.6]) and poor outcome at 6 months (odds ratio 2.5 [95% confidence interval 1.13-5.5]), respectively. DISCUSSION: BD could be used to predict the long-term morbidity and may not be related to morbidity and mortality at discharge. PMID- 20009678 TI - Pediatric abdominal injury patterns generated by lap belt loading. AB - BACKGROUND: Abdominal injuries can occur in children wearing adult seat belts during motor vehicle crashes. The particular anatomic structures injured may depend on the position of the seat belt, but this relationship has never been quantified in a controlled setting. METHODS: Thirty-four dynamic tests, designed to replicate crash loading, were performed on a validated porcine model of the 6 year-old human abdomen. The lap belt was positioned initially at either an "upper" or "lower" location on the abdomen. Detailed posttest necropsies identified all resulting injuries, which were then correlated to the initial position of the seat belt. The utility of using the presence or absence of an abdominal abrasion to predict internal abdominal injuries was also investigated. RESULTS: The injury frequency distributions from upper and lower abdominal belt loading were found to be significantly different (p < 0.01). Upper abdominal loading most commonly produced splenic lacerations and rib fractures, whereas lower abdominal loading resulted primarily in injuries to the large and small intestines. The presence of an abdominal abrasion was found to be a fairly specific (Sp = 0.86) but not very sensitive (Sn = 0.33) predictor of internal injuries of moderate severity. CONCLUSIONS: The relative frequency of each injury coupled with the presence or absence of an abdominal abrasion ("seat belt sign") should aid clinicians during their evaluations of children involved in motor vehicle crashes with possible occult abdominal trauma. In addition, the presence of a seat belt sign may have greater utility ruling in potentially occult abdominal injuries rather than excluding them. PMID- 20009679 TI - Selective nonoperative management of penetrating abdominal injuries in children. AB - BACKGROUND: Nonoperative management of solid organ injuries caused by blunt abdominal trauma has been the standard care for many years. However, operative management is considered the standard care for penetrating abdominal trauma by most surgeons. The aim of this study was to assess the feasibility of selective nonoperative management of penetrating abdominal trauma in children. METHODS: A total of 90 children suffering from penetrating abdominal trauma between 2003 and 2008 were evaluated. Patients who had hemodynamic instability or signs of bowel perforation underwent an immediate laparotomy. The remaining patients were observed with serial clinical examinations, radiologic evaluation, and hemoglobin level. RESULTS: There were 76 boys and 14 girls. The mean age was 9.9 years (range, 1-16 years). The mechanism of injury was stab wound in 60 patients (67%) and gunshot in 30 (33%). The most commonly injured organ was bowel (51.7%). Omentum or bowel was eviscerated through wound in seven patients; none of these patients had organ injury. Although 51 (56.6%) were treated nonoperatively, 39 patients (43.4%) required surgical treatment (19 of 60 stab wound, 20 of 30 gunshot). Of the 39 patients who underwent surgery, 6 (15.3%) were found to have no significant organ injury during surgery. Of the all, 51 patients who were initially followed nonoperatively, two patients required surgery. There were two complications. CONCLUSION: The majority of abdominal stab wound and many gunshot wounds can initially be managed nonoperatively in children, when there is no hemodynamic instability or signs of hollow viscus perforation. PMID- 20009680 TI - Practice patterns in the use of retrievable inferior vena cava filters in a trauma population: a single-center experience. AB - BACKGROUND: The use of permanent inferior vena cava filters (IVCFs) offers protection against pulmonary embolism (PE) but increases the long-term risk of deep vein thrombosis (DVT) and does not affect long-term mortality. The use of retrievable IVCFs in trauma patients offers the dual advantage of protection against PE during the risk period and the option of filter removal thus avoiding complications of DVT. Despite the safety of removal, it is likely that many of these retrievable filters are not removed. METHODS: This was a retrospective, single-center, observational cohort study at a rural level I trauma center. We sought to investigate the number of patients and the circumstances under which retrievable IVCFs were placed and removed. RESULTS: During a 4-year period, 3,455 trauma patients were admitted and 125 patients had retrievable IVCFs placed (71 therapeutic and 54 prophylactic). The most common indications were traumatic brain and spinal cord injuries (66%). During in-hospital filter use, there were 36 new incidences (29%) of PE (1) and DVT (35). Nine patients died before removal. In 40 patients (32%), removal was attempted, and 32 (26%) retrievable IVCFs were successfully removed and in most patients (76%) within 180 days of insertion. Seventeen patients were transferred out of the area for extended care and lost to follow-up. In 55 patients, the filters were not removed. In 20 patients, the surgeon decided against removal. Thirty patients were transferred to extended care or rehabilitation within the community, but they did not return for removal. Thus, of 108/125 patients with follow-up, 76 patients (70%) did not have their IVCFs removed, and 50 patients did not have their IVCFs removed because of the choice of the surgeon, extended care, or rehabilitation. CONCLUSIONS: The use of retrievable IVCFs, when necessary, produced predictable protection against PE and DVT complications. Despite the opportunity for removal, most patients, in fact, did not have their filters removed, even when posthospital care could be tracked. The practices of the surgeon, the transfer to extended-care facilities, near or far, and the reluctance to remove long-standing IVCFs contributed to the high-retention rate. PMID- 20009681 TI - Clinical examination in complement with computed tomography scan: an effective method for identification of cervical spine injury. AB - OBJECTIVE: The purpose of this study was to prospectively evaluate a protocol that assesses the efficacy and sensitivity of clinical examination in complement with computed tomographic (CT) scan in screening for cervical spine (c-spine) injury. METHODS: During the 26-month period from March 2005 to May 2007, blunt trauma patients older than 13 years were prospectively entered into a study protocol. If patients were awake and alert with Glasgow Coma Score (GCS) >or=14, clinical examination of the neck was performed. Clinical examination was performed regardless of distracting injuries. If the patient had no complaints of pain or tenderness, the cervical collar was removed. Patients with complaints of c-spine pain or tenderness and patients with GCS score <14 underwent CT scanning for evaluation of the entire c-spine. RESULTS: One thousand six hundred eighty seven patients were prospectively assessed for blunt c-spine injury. Fourteen hundred thirty-nine patients had GCS score >or=14, 897 (62%) of which had a negative clinical examination of the c-spine and subsequently had cervical collars removed. Two patients (0.2%) whose clinical examination results disclosed nothing abnormal were later found to have a c-spine injury. Five hundred forty two patients with GCS score >or=14 had a positive c-spine clinical examination, of which 134 (24%) were diagnosed with c-spine injury. One hundred thirty-three (99%) c-spine injuries were identified by CT scan. The c-spine injury missed by CT scan was a radiologic misinterpretation. For patients with c-spine injury with GCS score >or=14, both sensitivities of clinical examination and CT scan were 99%. Two hundred forty-eight patients had GCS score <14, of which 5 (2.0%) were diagnosed with c-spine injury. CT scan identified all c-spine injuries for patients with GCS score <14. CONCLUSIONS: In awake and alert blunt trauma patients, clinical examination is a sensitive screening method for c-spine injury. Clinical examination allows for the majority of blunt trauma patients to have their c-spines cleared safely without radiologic screening. Clinical examination in complement with CT scan is a sensitive and an effective method for identification of c-spine injury in awake and alert patients with symptoms of c spine injury. CT scan is the sensitive and effective test for screening and diagnosis of c-spine injury in blunt trauma patients with altered mental status. PMID- 20009682 TI - Clinical characterization of comatose patients with cervical spine injury and traumatic brain injury. AB - BACKGROUND: Reports on the risk factors for combined craniocervical spine injury in comatose patients are rare. The incidence of concomitant cervical injury in comatose patients with traumatic brain injury (TBI) was determined herein. METHODS: One thousand twenty-six comatose patients with TBI were examined. The clinical characteristics of combined craniocervical trauma were documented, including type and location of cervical injury, occurrence of hypotension, and dyspnea. RESULTS: Seventy-one patients (6.92%) sustained cervical spine injury. The most common injury region included the upper cervical segments, demonstrated in 37 (52.11%) of 71 patients. Of the 71 patients who sustained combined craniocervical spine injury, 42 (59.15%) had hypotension, including 26 (36.62%) with dyspnea. With regard to the association between the severity of TBI and the incidence of the cervical injury, a significant difference was apparent between patients with an initial Glasgow Coma Scale (GCS) score of 3-5 and those with an initial GCS score of 9-12 (11.62% compared with 4.03%, p < 0.01). Regarding the relationship between the mechanism of injury and the occurrence of cervical spine injury, cervical spine injury was associated at a significantly higher incidence with motorcycle accident-related head trauma as compared with non motorcycle accident-related trauma (10.32% vs. 4.68%, p < 0.01). CONCLUSION: Patients who sustained TBI as a result of motorcycle accidents and those exhibiting a lower GCS score are at the highest risk for concomitant cervical spine injury. PMID- 20009683 TI - Head injury as a PTSD predictor among Oklahoma City bombing survivors. AB - BACKGROUND: The aim of the Oklahoma City (OKC) bombing retrospective review was to investigate the relationship between physical injury, environmental contributors, and psychiatric disorders such as posttraumatic stress disorder (PTSD) in an event-based, matched design study focused on injury. METHODS: The 182 selected participants were a random subset of the 1,092 direct survivors from the OKC bombing. Only 124 of these 182 cases had a full complement of medical/clinical data in the OKC database. These 124 cases were assessed to explore relationships among PTSD diagnoses, levels of blast exposure, and physical injuries. Associations among variables were statistically tested using contingency analysis and logistic regression. RESULTS: Comparison of the PTSD cases to symptoms/diagnoses reported in the medical records reveals a statistically significant association between PTSD and head/brain injuries associated with head acceleration. PTSD was not highly correlated with other injuries. Although blast pressure and impulse were highly correlated with head injuries, the correlation with PTSD was not statistically significant. Thus, a correlation between blast pressure and PTSD may exist, but higher fidelity pressure calculations are required to elucidate this potential relationship. CONCLUSIONS: This study provides clear evidence that head injury is associated with subsequent PTSD, giving caregivers' information on what physical injuries may suggest the development of psychologic disorders to aid them in developing a profile for the identification of future survivors of terrorist attacks and Warfighters with brain injuries and potential PTSD. PMID- 20009684 TI - Ocular nail gun injuries: epidemiology and visual outcomes. AB - BACKGROUND: Nail-gun accidents have become a common reason for penetrating ocular injuries in patients presenting to the emergency room with globe trauma. To date, there are only five patients (one case report and one small case series) in the medical literature of penetrating ocular injury due to nail-guns. The aim of this study is to describe the epidemiology and visual outcomes of patients presenting to a tertiary medical center with ocular injuries resulting from nail-gun injuries. METHODS: This is an institutional review board-approved retrospective chart review of 24 patients presenting to the Medical University of South Carolina after sustaining eye injuries secondary to the use of nail-guns. The records of 178 patients with penetrating eye injuries treated between July 1996 and June 2006 were reviewed. Twenty-four patients (14%) had open globe injuries related to nail-guns. Demographic data, ocular examination findings, and visual outcomes were recorded. RESULTS: Of the 24 cases identified, 17 were work related; this represented approximately 71% of patients. All were men with an average age of 29.4 years. Fifty-eight percent of the patients were non-English speaking. There was no record of any patient wearing safety glasses at the time of injury. The visual acuity at presentation ranged from 20/20 to hand motion or worse. Half (50%) of these patients had a presenting visual acuity of hand motion or worse. At last examination, 38% (nine patients) had visual acuity of 20/40 or better, 34% (eight patients) had visual acuity of counting fingers to hand motion, 13% (three patients) 20/60, 4% (one patient) 20/200, and three were lost to follow-up. The correlation coefficient between visual acuity at presentation and most recent was significant at 0.679 (p = 0.001). CONCLUSIONS: The majority of nail-gun injuries presented in this series were work-related. These injuries could be prevented with adherence to established safety measures including proper training and education of nail-gun operators, the use of sequential-trip triggers, and required safety glasses. Visual acuity outcomes of these 24 patients are better than what might be expected due to the nature of the injury. PMID- 20009685 TI - Usefulness of ultrasonography in the treatment of nasal bone fractures. AB - BACKGROUND: Ultrasonography has recently been used in assessment and diagnosis of maxillofacial trauma because it is easy and quick to perform, inexpensive, portable, and noninvasive. The aim of this study was to estimate ultrasonography as an intraoperative repositioning control of nasal bone fractures. METHODS: We studied 32 patients with suspected nasal bone fracture. We performed preoperative computed tomography (CT) for evaluation of the type and extent of nasal bone fractures. We also took external photographs for evaluation of external deviation of the nose and nasal deformity. During surgery, we performed real-time ultrasonography-guided closed reduction using a 10 MHz linear transducer. After 1 year, we performed postoperative evaluation with CT and external photography. We classified patients into three groups according to their CT score. RESULTS: Patients were 23 males and 9 females aged 8-39 years. Clinical symptoms were pain, nasal swelling, nasal bleeding, and localized depression at the trauma site. In almost all patients, postoperative external photographs showed a symmetrical nasal dorsum without external deformity, and postoperative CT showed stabilization of bony fragments and good alignment of the nasal bone. Postoperatively, the CT score was 3 (excellent) in 25 patients, 2 (good) in 5 patients, and 1 (fair) in 2 patients. CONCLUSION: We suggest that ultrasonography is very useful for evaluating intraoperative repositioning of nasal bone fractures. PMID- 20009686 TI - Multiple intracranial carotid injuries: pitfalls in diagnosis by angiography and principles of endovascular treatment. AB - BACKGROUND: Simultaneous multiple intracranial carotid injuries (ICIs) after head trauma are rarely referred for treatment and are often times fatal. The purpose of this study was to describe the potential angiographic pitfalls in diagnosis of multiple ICIs and to report the principles of endovascular management in 15 patients with 34 ICIs. METHODS: A 12-year study of the 15 patients (8 men and 7 women) with 34 ICIs was completed, and patients were managed by endovascular treatment. Of the 34 ICIs, there were 22 traumatic carotid-cavernous fistulas (TCCFs), 6 traumatic carotid aneurysms, 5 meningeal arteriovenous fistulas, and 1 traumatic occlusion of carotid artery. Transarterial endovascular embolization was performed in 32 ICIs. RESULTS: Four traumatic carotid aneurysms, four meningeal arteriovenous fistulas, and a second hole of the TCCF were missed in early detection by initial cerebral angiograms. The causes of missed early detection of ICIs in angiograms were attributed to occur with TCCFs in the ipsilateral internal carotid artery territory due to overlooking (n = 4), overlap with nearby carotid artery and/or fistula drains of TCCFs (n = 2), steal phenomenon (n = 2), and a latent period of ICI (n = 1). Successful occlusion of 32 ICIs was achieved. On the modified Rankin scale applied in follow-up, 14 patients were assessed as stable clinical status. CONCLUSIONS: Early initial detection of ICIs in cerebral angiograms may be difficult if ICIs occur in the same carotid artery, particularly when they coexist with TCCF. However, as soon as TCCFs are occluded, postembolization angiograms should be scrutinized to find the potential associating ICIs, and endovascular management should be performed promptly. PMID- 20009687 TI - Risk factors for compartment syndrome in traumatic brachial artery injuries: an institutional experience in 139 patients. AB - BACKGROUND: The brachial artery is the most common vascular injury encountered in upper extremity trauma. If not treated promptly, it can result in compartment syndrome (CS) and long-term disability. Here, we report an institutional experience of traumatic brachial artery injuries and establish risk factors for the development of upper extremity CS in this setting. METHODS: A retrospective review of 139 patients with traumatic brachial artery injury from 1985 to 2001 at a single institution. Patients were divided into two cohorts, those with evidence of CS and those without CS (NCS), for comparison. RESULTS: One hundred thirty nine patients presented with traumatic brachial artery injuries (mean age, 28.4 years). Twenty-nine patients (20.9%) were diagnosed with upper extremity CS, and 28 of these patients underwent fasciotomy on recognition of their CS. Seven patients (6.4%) in the NCS cohort underwent fasciotomy as a result of absent distal pulses on initial examination. Mean follow-up was 51.6 days. Two patients required revision of their arterial repair, and one patient underwent amputation. The risk of CS was increased in the presence of combined arterial injuries (p = 0.03), combined nerve injuries (p = 0.04), motor deficits (p < 0.0001), fractures, and increased intraoperative blood loss (p = 0.001). Multivariate logistic regression performed on these variables revealed that elevated intraoperative blood loss, combined arterial injury, and open fracture were independent risk factors for the development of CS (OR 1.12, 5.79, and 2.68, respectively). CONCLUSION: Prompt evaluation and management of traumatic brachial artery injuries is important to prevent CS, which can lead to functional deficits. In the setting of combined arterial injury, open fracture, and significant intraoperative blood loss, prophylactic fasciotomy should be considered. PMID- 20009688 TI - Prostacyclin reduces elevation of intracranial pressure and plasma volume loss in lipopolysaccharide-induced meningitis in the cat. AB - BACKGROUND: Severe meningitis may compromise cerebral perfusion through increases in intracranial pressure (ICP) and through hypovolemia caused by a general inflammation with systemic plasma leakage. From its antiaggregative/antiadhesive and permeability-reducing properties, prostacyclin (PGI2) is a potential adjuvant treatment in meningitis, but previously published data have been ambiguous. The objective of this study was to evaluate the effects of PGI2 on meningitis on ICP, plasma volume, blood pressure, and cerebral oxidative metabolism. METHODS: Meningitis was induced by intrathecal injection of lipopolysaccharide (LPS, 0.8 x 10 units/kg) in cats. Four hours after the injection, the animals were randomized to intravenous treatment with either low-dose PGI2 (1 ng/kg/min) or the vehicle for 6 hours (n = 7 in each group). No LPS and no PGI2 or vehicle was given to three cats (sham group). Effects of treatment on ICP, mean arterial pressure, plasma volume (I-albumin technique), and brain tissue lactate/pyruvate ratio (microdialysis technique) were evaluated. RESULTS: ICP increased from 10.0 mm Hg +/- 1.3 mm Hg and 10.8 mm Hg +/- 1.7 mm Hg to 19.9 mm Hg +/- 1.7 mm Hg and 19.6 mm Hg +/- 3.3 mm Hg in the PGI2 and the vehicle group, respectively, 4 hours after the LPS injection (not significant). ICP increased further to 21.8 mm Hg +/ 4.5 mm Hg and to 25.8 mm Hg +/- 6.0 mm Hg after treatment for 6 hours with PGI2 or vehicle, respectively (p < 0.05). There was no significant difference in arterial pressure between groups. Plasma volume loss was less in the PGI2 group than in the vehicle group at the end of the experiment and urine production and arterial oxygenation was higher in the PGI2 group. Lactate/pyruvate ratio was within the normal range in all groups. CONCLUSION: Low-dose PGI2 may be a beneficial adjuvant therapy for meningitis by reducing elevation of ICP and plasma volume loss. PMID- 20009689 TI - Survey of national usage of trauma response charge codes: an opportunity for enhanced trauma center revenue. AB - BACKGROUND: The objective of this study was to survey Trauma Center (TC) members of the National Foundation for Trauma Care/Trauma Center Association of America to determine usage and consistency of trauma team response charge codes and critical care accommodation charges for severely injured patients. Potential over and underutilization of these enhanced reimbursements was assessed. METHODS: All TC members of the National Foundation for Trauma Care/Trauma Center Association of America were surveyed (2007) on usage of codes Universal Billing (UB) 68x; Field Locator (FL) 19 (now FL 14) patient type 5 "TC," UB 208 and Centers for Medicare and Medicaid Services codes G0390 and Ancillary Procedure Codes 0618. Data were collected on the use of 68x "Trauma Response" in combination with emergency room UB 450 Healthcare Common Procedure Coding System Critical Care E/M Level of Service 99291, as well as the daily accommodation (bed) charge code 208 for trauma critical care. RESULTS: We received 57 responses of 217 requests (response rate, 26.3%). Most responding TCs are charging for either full (86%) or partial (79%) trauma activation. Fewer are charging for trauma team evaluation fees (51%) and UB 208, trauma critical care accommodation code (33%). Charges are extremely variable between and across TC levels and among regions. Full trauma activation fees ranged from $837 to $24,964 with level II TCs charging more on average than level I TCs. As many as 63% of TCs failed to use or did not recognize combining codes 68x with ED 450 Healthcare Common Procedure Coding System 99291. CONCLUSION: Significant underused opportunities exist for enhanced revenue by improved implementation of trauma response codes. Wide ranges in charges and the low frequency of full implementation suggest that education and coordination are needed among hospital departments involved, as well as among the trauma care community at large, to realize optimal reimbursement for trauma care services. PMID- 20009690 TI - Transitional fractures of the distal tibia: a minimal access approach for osteosynthesis. AB - BACKGROUND: Transitional fractures typically occur in patients with partially closed growth plates. In case of displacements >2 mm, osteosynthesis is recommended. Open reduction and stabilization with lag screws is the most common approach. Infection and paraesthesia are common complications of this method. This study describes an alternative approach by closed reduction and cannulated screw fixation. METHODS: All patients with transitional fractures treated by closed reduction and cannulated screw fixation were identified. All patients with >1 year after implant removal were included in this investigation. Of 27 patients treated by this technique, 21 met the inclusion criteria and were sent a standardized questionnaire including the Foot Function Index (FFI). All cases with impairments in the FFI were invited for a follow-up examination. RESULTS: Postoperative complications occurred in two patients (8.3%): one reported paraesthesia of the hallux and one showed skin irritations above the screw insertion. Average follow-up time was 3.8 years after implant removal. Of 21 patients who returned the questionnaire, three patients (14.3%) had impairment in the FFI and were invited for clinical follow-up: one patient showed mild signs of osteoarthritis without visible gaps of the joint surface, one patient had minimally restricted pronation, and one had neither clinical nor radiologic correlates for the impairment reported. Sports activity was reduced in one patient. CONCLUSION: If closed reduction can be achieved fixation with cannulated screws proved to be a satisfactory method with little scars and good clinical results. PMID- 20009691 TI - The effect of nicotine on distraction osteogenesis: an experimental study on rabbits. AB - BACKGROUND: Tobacco smoke contains more than 4,000 constituents, but not all of them are reported to have adverse effects on bone healing after distraction osteogenesis. No research on the systemic administration of nicotine has been done on distraction osteogenesis of the weight-bearing long bones of the lower extremity. METHODS: Fourteen New Zealand white male rabbits underwent distraction osteogenesis on the right tibia and lengthening by 1 cm. Transdermal nicotine bands were applied onto the dorsal skin of the rabbits in the study group. Mineral density of the distraction zone was measured at weekly intervals using quantitative computerized tomography. Mechanical properties were assessed by torsional loading, and the regenerated bone tissue was subjected to histopathological examination. RESULTS: Comparisons of weekly measurements in both groups showed that while the increase in bone density in the nicotine group was higher, relative to the initial values, it was still far behind the average density obtained in the control group at the end of the experiment. Statistical analysis of mechanical data showed significant differences in the gradient of the regression lines and maximum torsional angles between the two groups. The histopathological assessments showed noticeable neovascularization in the study group, which was concluded to be a compensatory mechanism for the negative delaying effect of nicotine on bone healing. CONCLUSION: Systemic administration of nicotine can cause delays in the process of healing in distraction osteogenesis by its negative effect on the mineralization of the regenerate. Patients should be made aware of this negative impact of nicotine before the limb lengthening surgery. PMID- 20009692 TI - Prognostic significance of lower extremity long bone fractures after automobile versus pedestrian injuries. AB - BACKGROUND: Fractures of the tibia and femur are common after automobile versus pedestrian (AVP) injuries. This study evaluates the effect of age on the type of fracture and the incidence and type of associated injuries. METHODS: All patients involved in an AVP crash from January 1, 1995, through June 30, 2006, were included in the study. Variables obtained from our trauma registry included age, gender, Injury Severity Score, Abbreviated Injury Scale, intra-abdominal and spine injuries, and mortality. Patients were divided into three age groups for analysis: younger than 15 years, 15 years to 55 years, and older than 55 years. The incidence and type of associated injuries were assessed according to the presence or absence of fractures of the tibia only, femur only, or combined femur and tibia fractures. RESULTS: A total of 6,652 patients had AVP crashes and 1,936 (29%) had tibia or femur fractures (tibia, 20%; femur, 7%; tibia/femur, 3%). The incidence of tibia fractures increased with age ranging from 13% in those younger than 15 years to 25% in those older than 55 (p < 0.001). Combined tibia and femur fractures were also more common in adults and isolated femur fractures were more common in children. Hollow viscus injury was 1.8% in those with lower extremity fractures and 0.9% in those without (p = 0.0013). Mortality was 10% in those with fractures and 6% in those without (p < 0.0001) and was 20% in those with tibia and femur fractures. CONCLUSIONS: Age affects the incidence and type of lower extremity fractures after AVP injuries. The presence of lower extremity fracture is associated with a higher incidence of chest, spine, and intra-abdominal injuries. It is advisable that all patients with lower extremity fractures after AVP injuries be evaluated by a surgeon familiar with these injury patterns. PMID- 20009693 TI - Revision with dynamic compression plate and cancellous bone graft for aseptic nonunion after surgical treatment of humeral shaft fracture. AB - BACKGROUND: We evaluated the effect of revision with dynamic compression plate (DCP) and cancellous bone graft for aseptic nonunion after surgical treatments of humeral shaft fracture. METHOD: Eighty-six patients with aseptic nonunion of humeral shaft fracture after various surgical treatments were reviewed and analyzed retrospectively between January 1982 and August 2006. There were 59 men and 27 women with the average age of 42 years (range, 19-81 years). Thirty-one fractures were defined as atrophic nonunion, 45 fractures were hypertrophic nonunion, and 10 fractures could not be defined clearly. All the fractures were managed with removal of previous implants, open reduction and internal fixation with DCP, supplemented by cancellous bone graft. The follow-up period was 38 months in average (range, 12-288 months). Functional evaluations were done by Mayo Elbow Performance Index and the modified scale of Constant and Murley. RESULTS: All the nonunions united with the average union time of 18 weeks (range, 14-26 weeks). Complications included five temporary radial nerve palsies and two wound infections. In final follow-up, the shoulder and elbow functions of the operated limbs were all noted to be good or excellent. CONCLUSION: DCP with cancellous bone graft is a reliable and an effective treatment for revision of aseptic nonunion of humeral shaft fracture after surgical treatment. PMID- 20009694 TI - Is damage control orthopedics essential for the management of bilateral femoral fractures associated or complicated with shock? An animal study. AB - BACKGROUND: The maximum score of a single anatomic system, the Injury Severity Score, may not reflect the overall damage inflicted by bilateral femoral fractures and justify the strategy of damage control orthopedics (DCO). It is necessary to investigate effects of various therapeutic procedures on such fractures with or without shock to facilitate correct decision making on DCO. METHODS: A model of bilateral femoral fractures was made in 36 of 48 male New Zealand White rabbits. A model of bilateral femoral shaft fractures associated with shock was made. After resuscitation, a reamed intramedullary nailing fixation was performed in the first group (IM group), and an external fixation device applied in the second group (EF group), and the fractures in the third group (control group) were supported with splints only. They were divided into four groups: shock with IM nailing (shock-IM), shock with external fixation (shock-EF), shock with conservative method (shock-Cons), and intramedullary nailing without shock (nonshock-IM). Vital signs and inflammatory reactions were recorded. Thirty-six hours after the therapeutic procedures in four groups, the animals were killed for histologic evaluation. RESULTS: The changes of vital signs were most significant in shock-IM group (p < 0.05). The exaggerated levels of interleukin-6, Interleukin-10, and tumor necrosis factor alpha concentrations demonstrated a significant difference between all the groups-shock-IM and other groups (p < 0.05). As to histologic appearances, the statistical difference varies from organ to organ. There is highly significant difference when the IM group is compared with the other two groups as far as lungs are concerned. As to the liver, there is only significant difference between the IM group and the control group. In terms of kidney and heart, there is no significant difference cross the groups. As to histologic appearances, there is highly significant difference in lungs between shock-IM group and other three groups. There is significant difference in liver between the shock-IM group and the shock-Cons group (p < 0.05). Kidneys and heart were less affected cross the groups. CONCLUSIONS: In this study, an early reamed intramedullary nailing fixation procedure resulted in more adverse effects on system stress, inflammatory response, and multiple organs. The injuries also cause histologic damages to lungs and liver. Therefore, early reamed intramedullary nailing fixation may pose a potential risk of developing complications and adopting the DCO strategy may be more preferable. Shock and IM combined cause most severe damages, followed by IM without shock, shock plus EF, and shock plus conservative procedure in that order. If IM must be used for some reasons, it is desirable be delayed until shock has been fully controlled and vasculorespiratory stability restored. PMID- 20009695 TI - Video registration of trauma team performance in the emergency department: the results of a 2-year analysis in a Level 1 trauma center. AB - BACKGROUND: Trauma teams responsible for the first response to patients with multiple injuries upon arrival in a hospital consist of medical specialists or resident physicians. We hypothesized that 24-hour video registration in the trauma room would allow for precise evaluation of team functioning and deviations from Advanced Trauma Life Support (ATLS) protocols. METHODS: We analyzed all video registrations of trauma patients who visited the emergency room of a Level I trauma center in the Netherlands between September 1, 2000, and September 1, 2002. Analysis was performed with a score list based on ATLS protocols. RESULTS: From a total of 1,256 trauma room presentations, we found a total of 387 video registrations suitable for analysis. The majority of patients had an injury severity score lower than 17 (264 patients), whereas 123 patients were classified as multiple injuries (injury severity score >or=17). Errors in team organization (omission of prehospital report, no evident leadership, unorganized resuscitation, not working according to protocol, and no continued supervision of the patient) lead to significantly more deviations in the treatment than when team organization was uncomplicated. CONCLUSIONS: Video registration of diagnostic and therapeutic procedures by a multidisciplinary trauma team facilitates an accurate analysis of possible deviations from protocol. In addition to identifying technical errors, the role of the team leader can clearly be analyzed and related to team actions. Registration strongly depends on availability of video tapes, timely started registration, and hardware functioning. The results from this study were used to develop a training program for trauma teams in our hospital that specifically focuses on the team leader's functioning. PMID- 20009696 TI - The implementation of a national trauma registry in Greece. Methodology and preliminary results. AB - BACKGROUND: Trauma is a leading cause of death worldwide and a major health problem of the modern society. Trauma systems are considered the gold standard of managing patients with trauma. An integral part of any trauma system is a trauma registry. In Europe, and particularly in Greece, trauma registries and systems are in an embryonic stage. In this study, we present an attempt to record trauma in Greece. METHODS: The Hellenic Society of Trauma and Emergency Surgery invited all the official representatives of the society throughout the country to participate in the study. In succeeding meetings of the representatives, the reporting form was developed and the inclusion criteria were defined meticulously. Inclusion criteria were defined as patients with trauma requiring admission, transfer to a higher level center, or arrived dead or died in the emergency department of the reporting hospital. All reports were accumulated by the Hellenic Trauma society, imported in an electronic database, and analyzed. RESULTS: Thirty-two hospitals receiving patients with trauma participated in the country, representing 40% of the country's healthcare facilities and serving 40% of the country's population. In 12 months time, (October 2005 to September 2006), 8,862 patients were included in the study. Of them, 66.9% were men and 31.3% were women. The compilation rate of the reporting forms was surprisingly high, considering that the final reporting form included 150 data points and that there were no independent personnel in charge of filling the forms. CONCLUSIONS: Trauma registries are feasible even in health care systems where funding of medical research is sparse. PMID- 20009697 TI - Utility of the shock index in predicting mortality in traumatically injured patients. AB - BACKGROUND: Currently, specific triage criteria, such as blood pressure, respiratory status, Glasgow Coma Scale, and mechanism of injury are used to categorize trauma patients and prioritize emergency department (ED) and trauma team responses. It has been demonstrated in previous literature that an abnormal shock index (SI = heart rate [HR]/systolic blood pressure, >0.9) portends a worse outcome in critically ill patients. Our study looked to evaluate the SI calculated in the field, on arrival to the ED, and the change between field and ED values as a simple and early marker to predict mortality in traumatically injured patients. METHODS: A retrospective chart review of the trauma registry of an urban level I trauma center. Analysis of 2,445 patients admitted over 5 years with records in the trauma registry of which 1,166 also had data for the field SI. An increase in SI from the field to the ED was defined as any increase in SI regardless of the level of the magnitude of change. RESULTS: Twenty-two percent of patients reviewed had an ED SI >0.9, with a mortality rate of 15.9% compared with 6.3% in patients with a normal ED SI. An increase in SI between the field and ED signaled a mortality rate of 9.3% versus 5.7% for patients with decreasing or unchanged SI. Patients with an increase in SI of >or=0.3 had a mortality rate of 27.6% versus 5.8% for patients with change in SI of <0.3. CONCLUSION: Trauma patients with SI >0.9 have higher mortality rates. An increase in SI from the field to the ED may predict higher mortality. The SI may be a valuable addition to other ED triage criteria currently used to activate trauma team responses. PMID- 20009698 TI - Franz Weitlaner: the great spreader of surgery. PMID- 20009699 TI - Component separation technique for abdominal wall reconstruction in burn patients with decompressive laparotomies. AB - BACKGROUND: Component separation technique has been used successfully in ventral hernia repair occurring after damage control surgery. Abdominal compartment syndrome, seen in severely injured burn patients, frequently requires decompressive laparotomy. The patient is at risk during this time not only for burn injury complications but also for those from an open abdomen. METHODS: This report presents the successful application of the component separation technique for early closure of decompressive laparotomies in patients with >75% total body surface area burn, which included the abdominal wall. RESULTS: Skin flaps (necrotic/burned skin) overlying the abdominal wall fascia were raised bilaterally at the costal margin, from the anterior superior iliac spine inferiorly to the ribs superiorly. An incision was made just lateral to the rectus sheath through the aponeurosis of the external oblique muscle. With this, the fascia was mobilized to the middle with no tension. With no elevation of the patient's intrathoracic pressure on closure of the abdomen, multiple no. 2 Ethibond fascial figure of eight sutures closed the abdomen. Skin flaps were excised, so that grafting of the abdominal wall could occur. CONCLUSION: Burn patients, who required decompressive laparotomies for abdominal compartment syndrome in response to massive fluid resuscitation, tolerated early closure by the modified component separation technique. This markedly improved the care of these critically burned individuals, allowing for less third space fluid loss, less difficulty in management of the open abdominal wound, along with decreased risk of potential enterocutaneous fistula and intraabdominal abscess formation. PMID- 20009700 TI - Clinical practice guideline: red blood cell transfusion in adult trauma and critical care. PMID- 20009701 TI - Emergency innovation: implications for the trauma surgeon. AB - BACKGROUND: Experimentation involving human subjects requires careful attention to the protection of their rights. Beginning with the Belmont Report in 1979, the United States has developed various sets of rules and regulations that identify the requirements for performing human subject research. In addition, these standards attempt to define the fundamental difference between what constitutes research versus clinical treatment versus innovation. We explore the intersection between two areas of independent bioethics, surgical innovation and emergency research; the point we refer to as emergency innovation. METHODS: A systematic literature review in each of the fields of emergency research and surgical innovation was completed. The ethical principles involved in each field were identified. In addition, a recent case of surgical innovation within the context of emergency treatment is evaluated for the ethics invoked. RESULTS: One of the great challenges in emergency innovation is that the main protection offered in innovation (heightened informed consent) is not possible in the emergency context where in fact informed consent is waived. Interestingly, the rest of the protections outlined for each field are not mutually exclusive. They can and should be utilized in any project that takes place at this intersection. However, as there are no strict regulations in place for the collision of these two fields, the possibility of having the majority of the involved ethical principles misinterpreted or ignored is very real. CONCLUSIONS: For emergency innovation, where it is unclear what ethical principles and regulatory powers apply, it is imperative to be unambiguous about the purpose of the investigation, to adhere to all applicable ethical principles, and to have utmost consideration for protection of the research subject. To determine intent, the goals of the study must be outlined precisely - and if those include the prospect of publication, institutional review board (IRB) approval should be involved early. If, however, the innovation is subtle and the goal geared toward improved patient care, a small feasibility trial would be an appropriate first step before transitioning to a formal larger study approved by an IRB. In either case, the degree of the change in practice must be carefully evaluated and the vulnerability of the research subjects respected. With careful attention paid to all applicable ethical principles at the emergency innovation intersection, medical progress can continue at minimized risk to the human subject participants. PMID- 20009702 TI - Cerebral venous thrombosis associated with seat belt syndrome. PMID- 20009703 TI - Ruptured gallbladder: delayed presentation after motor vehicle collision. PMID- 20009704 TI - Differences in prescription of narcotic pain medication between hospitals in different countries. PMID- 20009705 TI - Economic policies and trauma. PMID- 20009710 TI - Temporary loss of perivascular aquaporin-4 in white matter after the spinal cord ischemic injury of rats. PMID- 20009746 TI - Residents' operative experience in general surgery programs. PMID- 20009747 TI - Ischemic complications after pancreatoduodenectomy: incidence, prevention and management. PMID- 20009748 TI - Management of severe acute pancreatitis: a surgical odyssey. AB - Throughout much of history, surgery of the pancreas was restricted to drainage of abscesses and treatment of traumatic wounds. At the turn of the 20th century under the impetus of anesthesia, such surgical stalwarts as Mayo Robson, Mickulicz, and Moynihan began to deploy laparotomy and gauze drainage in an effort to salvage patients afflicted with severe acute pancreatitis (SAP). Over the next thirty years, surgical intervention in SAP became the therapy for choice, despite surgical mortality rates that often exceeded 50%.When the discovery of the serum test for amylase revealed that clinically milder forms of acute pancreatitis existed that could respond to nonoperative therapy, a wave of conservatism emerged, and, for the next quarter century, surgical intervention for SAP was rarely practiced. However, by the 1960s, conservative mortality rates for SAP were reported to be as high as 60% to 80%, leading surgeons to not only refine the indications for surgery in SAP, but also to consider new approaches. Extensive pancreatic resections for SAP became the vogue in continental surgical centers in the 1960s and 1970s, but often resulted in high mortality rates and inadvertent removal of viable tissue.Accurate diagnosis of pancreatic necrosis by dynamic CT led to new approaches for management. Some surgeons recommended restricting intervention to those with documented infected necrosis, and proposed delayed exploration employing sequestrectomy and open-packing. Others advocated debridement early in the course of the disease for all patients with necrotizing pancreatitis, regardless of the status of infection. In the 1990s, however, a series of prospective studies emerged proving that nonoperative management of patients with sterile pancreatic necrosis was superior to surgical intervention, and that delayed intervention provided improved surgical mortality rates.The surgical odyssey in managing the necrotizing form of SAP, from simple drainage, to resection, to debridement, to sequestrectomy, although somewhat tortuous, is nevertheless an notable example of how evidence-based knowledge leads to improvement in patient care. Today's 10% to 20% surgical mortality rates reflect not only considerable advances in surgical management, but also highlight concomitant improvements in fluid therapy, antibiotics, and intensive care. Although history documents the important contributions that surgical practitioners have made to acute pancreatitis and its complications, surgeons are rarely complacent, and the recent emergence of minimally invasive techniques holds future promise for patients afflicted with this "... most formidable of catastrophes." PMID- 20009749 TI - Predictors of the presence of concomitant invasive ductal carcinoma in intraductal papillary mucinous neoplasm of the pancreas. AB - OBJECTIVE: Invasive ductal carcinoma (DC) of the pancreas arising as an independent lesion in association with intraductal papillary mucinous neoplasm (IPMN) has occasionally been reported. However, clinicopathological features related to the presence of DC in patients with IPMN remain largely unknown. The purpose of the present study was to determine the factors predicting the presence of concomitant DC in those with IPMN. MATERIALS AND METHODS: We retrospectively reviewed the clinicopathological data of a consecutive series of 236 patients with IPMN treated by surgical resection or followed up at our institution between January 1987 and June 2008. In an attempt to identify predictors for the presence of DC, clinicopathological variables were compared between IPMN patients with concomitant DC and those without concomitant DC. RESULTS: Of 236 patients with IPMN, concomitant DC was detected synchronously or metachronously in 22 patients (9.3%). All the 22 IPMNs were of branch duct type and histological grades of 12 resected IPMNs were adenoma(n = 8) and borderline (n = 4). Multivariate analysis revealed 2 significant predictive factors for the presence of DC in IPMN, including worsening diabetes mellitus (P < 0.001) and an abnormal serum CA 19-9 level (P = 0.024). CONCLUSION: In view of the high prevalence of DC careful inspection of the entire pancreatic gland is necessary for early detection of DC in patients with branch duct IPMNs, especially when worsening diabetes mellitus and an abnormal serum CA 19-9 level are manifested. PMID- 20009750 TI - The influence of mechanical bowel preparation in elective lower colorectal surgery. AB - OBJECTIVE: This study evaluates the effects of mechanical bowel preparation (MBP) on anastomosis below the peritoneal verge and questions the influence of MBP on anastomotic leakage in combination with a diverting ileostomy in lower colorectal surgery. SUMMARY BACKGROUND DATA: In a previous large multicenter randomized controlled trial MBP has shown to have no influence on the incidence of anastomotic leakage in overall colorectal surgery. The role of MBP in lower colorectal surgery with or without a diverting ileostomy remains unclear. METHODS: This study is a subgroup analysis of a prior multicenter (13 hospitals) randomized trial comparing clinical outcome of MBP versus no MBP. Primary end point was the occurrence of anastomotic leakage and secondary endpoints were septic complications and mortality. RESULTS: Total of 449 Patients underwent a low anterior resection with a primary anastomosis below the peritoneal verge. The incidence of anastomotic leakage was 7.6% for patients who received MBP and 6.6% for patients who did not. Significant risk factors for anastomotic leakage were the American Society of Anesthesiologists-classification (P = 0.005) and male gender (P = 0.007). Of total, 48 patients received a diverting ileostomy during initial surgery; 27 patients received MBP and 21 patients did not. There were no significant differences regarding septic complications and mortality between both groups. CONCLUSION: MBP has no influence on the incidence of anastomotic leakage in low colorectal surgery. Furthermore, omitting MBP in combination with a diverting ileostomy has no influence on the incidence of anastomotic leakage, septic complications, and mortality rate. PMID- 20009751 TI - Expansion of a cell population expressing stem cell markers in parathyroid glands from patients with hyperparathyroidism. AB - OBJECTIVE: We sought to examine abnormal parathyroid glands for the presence of stem cells. SUMMARY BACKGROUND DATA: Cancer stem cells have been identified in cancers from a variety of tissues as a CD44/CD24 cell population. We hypothesize that stem cells (SC) may also be involved in the pathogenesis of benign clonal expansion characteristic of hyperparathyroidism (HPT). METHODS: Under institutional review board approval, parathyroid tissue was obtained from 20 patients with HPT and analyzed by fluorescence-activated cell sorting (FACS) for the CD44/CD24 cell population. Immunohistochemistry (IHC) with CD44 antibody was correlated with FACS results. RESULTS: Parathyroid tissue was obtained for FACS analysis from 25 enlarged parathyroid glands from 20 patients, 17 with primary HPT, and 3 with secondary HPT. The average percent of SC defined as CD44/CD24 population was 10.93% for enlarged parathyroid glands. IHC using CD44 antibody was performed on 27 abnormal parathyroid glands and 7 normal parathyroid gland biopsies from the same patients. Although IHC was not as sensitive as FACS, comparison of IHC and FACS results for 24 abnormal glands gave a correlation coefficient of 0.52, which was statistically significant (P = 0.01, Spearman rank). By IHC, 13 of 27 abnormal glands stained 1+ to 3+ (average, 0.93) compared with no CD44 staining in normal glands, which was statistically different (mean IHC of 0 vs. 0.93, P = 0.03, Wilcoxon). CONCLUSIONS: These novel findings demonstrate expansion of a resident cell population that expresses SC markers in abnormal parathyroid glands from patients with HPT. Our results suggest that clonal expansion of a resident SC population occurs in the pathogenesis not only of cancer, but also in benign parathyroid tumors occurring in HPT. PMID- 20009752 TI - The Agency for Healthcare Research and Quality (AHRQ) pediatric quality indicators (PDIs): accidental puncture or laceration during surgery in children. AB - CONTEXT: The Agency for Healthcare Research and Quality (AHRQ) pediatric quality indicators (PDIs) are measures designed to evaluate the quality of pediatric healthcare. They specifically focus on adverse events that are potentially avoidable, including complications and iatrogenic events. PDI 1 refers to accidental puncture or laceration. OBJECTIVE: To determine risk factors and outcomes associated with PDI 1 in a population of pediatric surgical patients. DESIGN, SETTING, AND PATIENTS: The Nationwide Inpatient Sample and Kids Inpatient Database were used to identify hospitalized pediatric surgical patients in the United States (age: 0-18) from 1988 to 2005. The data from these 1,939,540 patients was linked to the AHRQ PDIs using AHRQ WinQI software, and 7,033 pediatric patients with PDI 1 were identified. A 1:3 matched case control design was implemented with 6,459 cases (patients with PDI 1) and 19,377 controls (patients without PDI 1) matched on age, race, gender, and hospital ID. Cases and controls were stratified into procedure categories based on diagnosis related group procedure codes. MAIN OUTCOME MEASURES: To examine the relationship between PDI 1 and procedure category, as well as the outcomes of in-hospital mortality, length of stay, and total hospital charges for cases compared with controls. RESULTS: Of the 4,627 patients with PDI 1 stratified into procedure categories, the highest proportion of PDI 1 cases occurred in the gastrointestinal (30.19%), cardiothoracic (19.6%), and the orthopedic (11.13%) categories. Logistic regression analysis for PDI 1, controlling for admission type and insurance status, revealed a statistically significant higher odds of PDI 1 in the gynecology (OR: 1.69, P < 0.001) and transplant (OR: 1.45, P: 0.026) procedure categories. Multivariable regression analysis revealed patients with PDI 1 were more likely to die (OR: 1.91, P < 0.001), had a 4.81 day longer length of stay (95% CI: 4.26-5.36, P < 0.001) and had USD 36,291 higher total hospital charges (95% CI: USD 32,583-USD 40,000, P < 0.001) compared with patients without PDI 1. CONCLUSIONS: Cases of PDI 1 were most commonly associated with the gastrointestinal, cardiothoracic, and orthopedic procedure categories, and these were also 3 of the most common procedure categories overall. Controlling for type of procedure and other variables, the procedure categories having the highest likelihood of PDI 1 were gynecology and transplant. PDI 1 was found to be associated with greater mortality, longer length of stay, and greater total hospital charges. PMID- 20009753 TI - Trends and outcomes of hospitalizations for peptic ulcer disease in the United States, 1993 to 2006. AB - OBJECTIVES: Despite progress in diagnosis and treatment, peptic ulcer disease (PUD) remains a common reason for hospitalization and operation. The purpose of this study was to quantify the time trends of hospitalizations and operations for PUD in the United States (US) since 1993. DATA AND METHODS: The Healthcare Cost and Utilization Project Nationwide Inpatient Sample is a 20% stratified sample of all hospitalizations in the United States. It was used to study hospitalizations with PUD as the principal diagnosis during 1993 to 2006, including details on ulcer site, complications, procedures, and mortality. Statistical methods included the chi test and multivariate logistic regression. RESULTS: The national estimate of hospitalizations for PUD decreased significantly from 222,601 in 1993 to 156,108 in 2006 (-29.9%), with a larger reduction in duodenal ulcers (95,552 in 1993 vs. 60,029 in 2006, -37.2%) than gastric ulcers (106,987 in 1993 vs. 86,064 in 2006, -19.6%). The inpatient mortality rate of PUD decreased from 3.8% to 2.7% during 1993 to 2006 (P < 0.001). Hemorrhage remained the most common complication (71.6% in 1993; 73.3% in 2006) but perforation had the highest mortality (15.1% in 1993; 10.6% in 2006). In comparison to 1993, patients hospitalized for PUD in 2006 more frequently had endoscopic treatment to control bleeding (12.9% vs. 22.2%, P < 0.001), similar use of surgical oversewing of ulcer (7.6% vs. 7.4%), less use of gastrectomy (4.4% vs. 2.1%, P < 0.001), and less use of vagotomy (5.7% vs. 1.7%, P < 0.001). In multivariate logistic regressions, the determinants of mortality were similar in 1993 and 2006. CONCLUSIONS: Hospitalizations for PUD decreased in the United States from 1993 to 2006, suggesting a decrease in the prevalence and/or severity of ulcer complications over this recent time period. Despite increased patient age and comorbidities, there has been a significant decrease in PUD mortality, a significant increase in the use of therapeutic endoscopy for bleeding ulcer, and a significant decrease in the use of definitive surgery (vagotomy or resection) for ulcer complications. PMID- 20009754 TI - The evolution of the surgical treatment of chronic pancreatitis. AB - OBJECTIVE: To establish the current status of surgical therapy for chronic pancreatitis, recent published reports are examined in the context of the historical advances in the field. BACKGROUND: The basis for decompression (drainage), denervation, and resection strategies for the treatment of pain caused by chronic pancreatitis is reviewed. These divergent approaches have finally coalesced as the head of the pancreas has become apparent as the nidus of chronic inflammation. METHODS: The recent developments in surgical methods to treat the complications of chronic pancreatitis and the results of recent prospective randomized trials of operative approaches were reviewed to establish the current best practices. RESULTS: Local resection of the pancreatic head, with or without duct drainage, and duodenum-preserving pancreatic head resection offer outcomes as effective as pancreaticoduodenectomy, with lowered morbidity and mortality. Local resection or excavation of the pancreatic head offers the advantage of lowest cost and morbidity and early prevention of postoperative diabetes. The late incidences of recurrent pain, diabetes, and exocrine insufficiency are equivalent for all 3 surgical approaches. CONCLUSIONS: Local resection of the pancreatic head appears to offer best outcomes and lowest risk for the management of the pain of chronic pancreatitis. PMID- 20009755 TI - Cancer causes increased mortality and is associated with altered apoptosis in murine sepsis. AB - OBJECTIVE: Whereas most septic patients have an underlying comorbidity, most animal models of sepsis use mice that were healthy before the onset of infection. Malignancy is the most common comorbidity associated with sepsis. The purpose of this study was to determine whether mice with cancer have a different response to sepsis than healthy animals. DESIGN: Prospective, randomized controlled study. SETTING: Animal laboratory in a university medical center. SUBJECTS: C57Bl/6 mice. INTERVENTIONS: Animals received a subcutaneous injection of either 250,000 cells of the transplantable pancreatic adenocarcinoma cell line Pan02 (cancer) or phosphate-buffered saline (healthy). Three weeks later, mice given Pan02 cells had reproducible, nonmetastatic tumors. Both groups of mice then underwent intratracheal injection of either Pseudomonas aeruginosa (septic) or 0.9% NaCl (sham). Animals were killed 24 hrs postoperatively or followed-up 7 days for survival. MEASUREMENTS AND MAIN RESULTS: Mice with cancer and healthy mice appeared similar when subjected to sham operation, although cancer animals had lower levels of T- and B-lymphocyte apoptosis. Septic mice with cancer had increased mortality compared to previously healthy septic mice subjected to the identical injury (52% vs. 28%; p = .04). This was associated with increased bacteremia but no difference in local pulmonary infection. Septic mice with cancer also had increased intestinal epithelial apoptosis. Although sepsis induced an increase in T- and B-lymphocyte apoptosis in all animals, septic mice with cancer had decreased T- and B-lymphocyte apoptosis compared to previously healthy septic mice. Serum and pulmonary cytokines, lung histology, complete blood counts, and intestinal proliferation were similar between septic mice with cancer and previously healthy septic mice. CONCLUSIONS: When subjected to the same septic insult, mice with cancer have increased mortality compared to previously healthy animals. Decreased systemic bacterial clearance and alterations in intestinal epithelial and lymphocyte apoptosis may help explain this differential response. PMID- 20009756 TI - Do hospitals provide lower quality of care to black patients for pneumonia? AB - OBJECTIVES: Recent studies reported lower quality of care for black vs. white patients with community-acquired pneumonia and suggested that disparities persist at the individual hospital level. We examined racial differences in emergency department and intensive care unit care processes to determine whether differences persist after adjusting for case-mix and variation in care across hospitals. DESIGN: Prospective, observational cohort study. SETTING: Twenty-eight U.S. hospitals. PATIENTS: Patients with community-acquired pneumonia: 1738 white and 352 black patients. INTERVENTIONS: None. MEASUREMENTS: We compared care quality based on antibiotic receipt within 4 hrs and adherence to American Thoracic Society antibiotic guidelines, and intensity based on intensive care unit admission and mechanical ventilation use. Using random effects and generalized estimating equations models, we adjusted for case-mix and clustering of racial groups within hospitals and estimated odds ratios for differences in care within and across hospitals. MAIN RESULTS: Black patients were less likely to receive antibiotics within 4 hrs (odds ratio, 0.55; 95% confidence interval, 0.43-0.70; p < .001) and less likely to receive guideline-adherent antibiotics (odds ratio, 0.72; 95% confidence interval, 0.57-0.91; p = .006). These differences were attenuated after adjusting for casemix (odds ratio, 0.59; 95% confidence interval; 0.46-0.76 and 0.84; 95% confidence interval, 0.66 -1.09). Within hospitals, black and white patients received similar care quality (odds ratio, 1; 95% confidence interval, 0.97-1.04 and 1; 95% confidence interval, 0.97 1.03). However, hospitals that served a greater proportion of black patients were less likely to provide timely antibiotics (odds ratio, 0.84; 95% confidence interval, 0.78-0.90). Black patients were more likely to receive mechanical ventilation (odds ratio, 1.57; 95% confidence interval, 1.02-2.42; p = .042). Again, within hospitals, black and white subjects were equally likely to receive mechanical ventilation (odds ratio, 1; 95% confidence interval, .94-1.06) and hospitals that served a greater proportion of black patients were more likely to institute mechanical ventilation (odds ratio, 1.13; 95% confidence interval, 1.02 1.25). CONCLUSIONS: Black patients appear to receive lower quality and higher intensity of care in crude analyses. However, these differences were explained by different case-mix and variation in care across hospitals. Within the same hospital, no racial differences in care were observed. PMID- 20009757 TI - Low-molecular-weight polyethylene glycol improves survival in experimental sepsis. AB - OBJECTIVE: For several chronic inflammatory disease states, therapy is enhanced by improving the pharmacokinetic properties of anti-inflammatory drugs through conjugation with polyethylene glycol. We hypothesized that part of the beneficial action of PEGylated drugs may be derived from the anti-inflammatory properties of polyethylene glycol (PEG) itself. DESIGN: Randomized, double-blinded, controlled ex vivo and in vivo laboratory studies. SETTING: University research laboratories. SUBJECTS: Human neutrophils and mononuclear cells, macrophage cell line, and adult rats and mice. INTERVENTIONS: The effect of PEG (either low molecular-weight [200-400] or high-molecular-weight [>4000]) was assessed on survival after systemic inflammation induced by lipopolysaccharide or zymosan. The effects of PEG on zymosan, lipopolysaccharide, or streptolysin-induced inflammatory and bioenergetic responses of immune cells were also assessed. MEASUREMENTS AND MAIN RESULTS: Low-molecular-weight PEG reduced inflammatory cytokine expression, pyrexia, and mortality by >50% in both lipopolysaccharide and zymosan models of sepsis. Low-molecular-weight PEG reduced cytokine expression both in vivo and in vitro, and attenuated activation of human neutrophils in response to lipopolysaccharide or zymosan. By contrast, high molecular-weight PEG conferred less significant survival effects after lipopolysaccharide and zymosan, and it did not exhibit such profound anti inflammatory effects. Low-molecular-weight PEG attenuated lipopolysaccharide induced activation of pro-apoptotic pathways (lysophosphatidic acid receptor and caspase-domain signaling) in the livers of endotoxemic rats. Streptolysin-induced necrosis of human neutrophils was reduced by low-molecular-weight PEG, indicating a mechanism that involves coating and/or stabilizing the cellular membrane. Low molecular-weight PEG preserved human neutrophil responses to septic serum and bioenergetic function in macrophages and neutrophils. CONCLUSION: PEG is a commonly used, safe, nonimmunogenic molecule possessing hitherto unappreciated anti-inflammatory properties. Low-molecular-weight PEG may potentially play a role in the therapy of systemic inflammation and sepsis. PMID- 20009758 TI - Continuous administration of pyridostigmine improves immobilization-induced neuromuscular weakness. AB - OBJECTIVE: To investigate the effects of continuous pyridostigmine infusion on immobilization-induced muscle weakness. Critical illness often results in immobilization of limb and respiratory muscles, leading to muscle atrophy and up regulation of nicotinic acetylcholine receptors. Pyridostigmine reversibly blocks acetylcholinesterase and has the potential to improve neuromuscular transmission and decrease acetylcholine receptor number. DESIGN: Prospective, randomized, controlled experimental study. SETTING: Animal laboratory, university hospital. SUBJECTS: Male Sprague-Dawley rats. INTERVENTIONS: A total of 40 rats were immobilized in one hind limb by pinning knee and ankle joints. Rats received either continuous pyridostigmine (15 mg.kg.day) or saline subcutaneously via implanted osmotic pumps. MEASUREMENTS AND MAIN RESULTS: After 7 days and 14 days of immobilization, neuromuscular function, atracurium pharmacodynamics, and expression of acetylcholine receptors were evaluated. At 7 days and 14 days after immobilization, muscle force decreased in all untreated groups, whereas effective doses for paralysis with atracurium and acetylcholine receptor number in the tibialis were significantly increased. Pyridostigmine-treated rats showed a significantly improved muscle force and muscle mass in the immobilized limb. This was associated with an attenuation of acetylcholine receptor up-regulation in the respective leg. At this time, the dose-response curve for atracurium on the immobilized side was shifted to the left in the pyridostigmine group. After 14 days, muscle tension was still less depressed with pyridostigmine infusion, and resistance to the effects of atracurium was still attenuated. However, there were no differences in acetylcholine receptor expression between the immobilized sides of both groups. CONCLUSIONS: Continuous pyridostigmine infusion improves muscle weakness after 7 days and 14 days of immobilization. The up-regulation of acetylcholine receptors and the concomitant resistance to atracurium is attenuated in animals treated with pyridostigmine after 7 days of immobilization. PMID- 20009759 TI - The potential successes and challenges of targeted anticancer therapies. PMID- 20009760 TI - A physiological approach to modernize the management of cancer chemotherapy induced gastrointestinal toxicity. AB - PURPOSE OF REVIEW: Gastrointestinal toxicity during chemotherapy is frequent. Symptomatic therapies for gastrointestinal toxicity, which do not address the underlying cause, may result in inadequate symptom control. With advances in curative treatment regimens, it becomes more important to minimize treatment toxicity which otherwise may compromise optimal chemotherapy and the chance of cure. RECENT FINDINGS: For decades, oncologists have concentrated on delineating the pathological processes, which occur within the gastrointestinal tract during chemotherapy treatment. However, pathological change does not in itself cause symptoms. Symptoms only arise when physiological functions are altered. In immunosuppressed patients, it is a priority to exclude infection as a cause for symptoms. In the presence of diarrhoea, the best investigative paradigm for this is stool culture, upper gastrointestinal endoscopy with duodenal biopsies and duodenal aspirate combined with flexible sigmoidoscopy and left colonic biopsies. Once infection has been excluded, although large studies have not been performed, case series repeatedly suggest that gastrointestinal symptoms arising during cancer chemotherapy can often be cured if newly acquired, gastrointestinal physiological deficits are identified. SUMMARY: Although many physiological changes induced by chemotherapy may be relatively transient and settle weeks or months after the end of treatment, during chemotherapy, their manifestations can be protean and severe. A systematic failure to research the frequency with which physiological causes of the symptoms are amenable to treatment has denied many patients' logical therapies for their chemotherapy-induced symptoms, and as a result, it is likely that symptomatic treatments are frequently suboptimal. PMID- 20009761 TI - High prevalence of array comparative genomic hybridization abnormalities in adults with unexplained intellectual disability. AB - PURPOSE: Array comparative genomic hybridization is now a widely used clinical tool for the evaluation of intellectual disability. The current 10% yield of positive findings is based largely on pediatric data. Adults with unexplained intellectual disability have not been systematically studied with array comparative genomic hybridization. Here, we report our initial experience with array comparative genomic hybridization testing on 45 adults with unexplained intellectual disability referred to an adult genetics clinic. METHODS: Beginning in 2006, we applied clinically available array comparative genomic hybridization testing to adults referred with an intellectual disability phenotype. The initial platform used was an early generation targeted or constitutional array, which was replaced by our current platform using more than 5000 bacterial artificial chromosome clones with an average resolution of 500 Kb and targeting 114 disease loci. All patients also underwent high-resolution karyotype analysis and molecular testing for Fragile X syndrome. RESULTS: Our population comprised 45 patients with unexplained intellectual disability (18 men and 27 women) with an average age of 35.1 years. Most patients had not been evaluated by genetics clinics since childhood or had never undergone a genetic evaluation; only two had documentation of prior normal karyotype studies. Three subjects had abnormal high resolution chromosome studies, which were also confirmed by array comparative genomic hybridization. Seven of the remaining 42 patients (17%) had novel genomic losses identified only by array comparative genomic hybridization. CONCLUSION: Abnormal genomic losses detected by array comparative genomic hybridization are prevalent in adults with unexplained intellectual disability. Our data showing abnormalities in 22% and 17% of overall patients and of cases with normal karyotypes, respectively, suggest that the yield of array comparative genomic hybridization in adults with unexplained intellectual disability may be higher than in pediatric populations. PMID- 20009762 TI - New roles of EDNRB and EDN3 in the pathogenesis of Hirschsprung disease. AB - PURPOSE: Hirschsprung disease is characterized by the absence of intramural ganglion cells in the myenteric and submucosal plexuses within distal intestine, because of a fail in the enteric nervous system formations process. Endothelin-3 endothelin receptor B signaling pathway is known to play an essential role in this process. The aim of this study was to evaluate the implication of the EDN3 and EDNRB genes in a series of patients with Hirschsprung disease from Spain and determinate their mutational spectrum. METHODS: We performed the mutational screening of both genes in 196 patients with Hirschsprung disease using denaturing high-performance liquid chromatography technology. A case-control study using TaqMan Technology was also carried out to evaluate some common polymorphisms and haplotypes as susceptibility factors for Hirschsprung disease. RESULTS: Besides several novel mutations in both genes, we found a truncating mutation in an alternative isoform of EDNRB. Interestingly, we obtained an overrepresentation of a specific EDN3 haplotype in cases versus controls. CONCLUSIONS: Our results suggest that the isoform EDNRB Delta 3 might be playing an essential role in the formation of enteric nervous system. In addition, based on the haplotype distribution, EDN3 might be considered as a common susceptibility gene for sporadic Hirschsprung disease in a low-penetrance fashion. PMID- 20009764 TI - Use of HIV resistance testing after prolonged treatment interruption. AB - BACKGROUND: HIV-1 genotypic resistance testing is not routinely recommended for patients who have been off antiretroviral therapy (ART) for longer than 4 weeks. We assessed the results and use of resistance testing in patients off ART. METHODS: All HIV resistance genotypes from November 2003 through April 2008 were reviewed from one large teaching hospital and two private HIV practices. Inclusion criterion was having a genotypic resistance test after an ART interruption of at least 2 months. Medical records were reviewed using a standardized data collection sheet. RESULTS: Sixty-two of 304 treatment experienced patients with HIV genotypes met the inclusion criteria. Prior cumulative ART class exposure included nucleoside reverse transcriptase inhibitors in 54 patients, nonnucleoside reverse transcriptase inhibitors in 32 patients, and protease inhibitors in 30 patients. Resistance testing was performed at a mean of 12 months (range, 2.5-48 months) after ART interruption. The mean time between ART interruption and resistance testing did not differ for patients with mutations and those without mutations detected. Seventeen of 62 (27.4%) patients were found to have resistance mutations. Eleven patients were found to have mutations to nonnucleoside reverse transcriptase inhibitors, four patients had mutations to nucleoside reverse transcriptase inhibitors, and two patients had protease inhibitor-associated mutations. No patient had multiclass resistance. Among the 17 patients with mutations after treatment interruption, 15 had mutations that were either not present on a prior genotype (n = 2) or did not have a prior genotype (n = 13). CONCLUSIONS: HIV genotypic resistance assays may identify mutations even when performed after a prolonged treatment interruption and may offer clinically significant information. Current guidelines that discourage resistance testing after treatment interruptions of longer than 4 weeks should be re-evaluated. PMID- 20009765 TI - Early clinical and programmatic outcomes with tenofovir-based antiretroviral therapy in Zambia. AB - BACKGROUND: In July 2007, amid some controversy over cost, Zambia was the first African country to introduce tenofovir (TDF) as a component of first-line antiretroviral therapy (ART) on a wide scale. METHODS: We compared drug substitutions, mortality, and "programmatic failure" among adults starting TDF-, zidovudine (ZDV)-, and stavudine (d4T)-containing ART. Programmatic failure was defined as death, withdrawal, or loss to follow-up. RESULTS: Between July 2007 and January 2009, 10,485 adults initiated ART (66% on TDF, 23% on ZDV, 11% on d4T), with a median follow-up time of 239 (interquartile range 98, 385) days. Those starting TDF were more likely to be male and more likely to have indicators of severe disease at baseline. In adjusted Cox proportional hazards models, ZDV- (adjusted hazard ratio [AHR] = 2.74, 95% confidence interval [CI] = 2.30-3.28) and d4T-based regimens (AHR = 1.92, 95% CI = 1.55-2.38) were associated with higher risk for drug substitution when compared with TDF-based regimens. Similar hazards were noted for overall mortality (ZDV: AHR = 0 .81, 95% CI = 0.62-1.06; d4T: AHR = 1.03, 95% CI = 0.74-1.43) and programmatic failure (ZDV: AHR = 0.99, 95% CI = 0.88-1.11; d4T: AHR = 1.11, 95% CI = 0.96-1.28) when compared with TDF. CONCLUSIONS: TDF is associated with similar clinical and programmatic outcomes as ZDV and d4T but appears to be better tolerated. Although further evaluation is needed, these results are encouraging and support Zambia's policy decision. PMID- 20009766 TI - The association between alcohol consumption and prevalent cardiovascular diseases among HIV-infected and HIV-uninfected men. AB - OBJECTIVE: To determine whether alcohol consumption is associated with cardiovascular disease (CVD) among HIV-infected veterans. METHODS: Using established thresholds for alcohol consumption, we analyzed cross-sectional data from 4743 men (51% HIV infected) from the Veterans Aging Cohort Study, a prospective cohort of HIV-infected veterans and demographically similar HIV uninfected veterans. Using logistic regression, we estimated the odds ratio (OR) for the association between alcohol consumption and prevalent CVD. RESULTS: Among HIV-infected and HIV-uninfected men, respectively, hazardous drinking (33.2% vs. 30.9%,), alcohol abuse and dependence (20.9% vs. 26.2%), and CVD (14.6% vs. 19.8%) were common. Among HIV-infected men, hazardous drinking [OR = 1.43, 95% confidence interval (CI) = 1.05 to 1.94] and alcohol abuse and dependence (OR = 1.55, 95% CI = 1.07 to 2.23) were associated with a higher prevalence of CVD compared with infrequent and moderate drinking. Among HIV-uninfected men, past drinkers had a higher prevalence of CVD (OR = 1.30, 95% CI = 1.01 to 1.67). For HIV-infected and HIV-uninfected men, traditional risk factors and kidney disease were associated with CVD. CONCLUSIONS: Among HIV-infected men, hazardous drinking and alcohol abuse and dependence were associated with a higher prevalence of CVD compared with infrequent and moderate drinking even after adjusting for traditional CVD risk factors, antiretroviral therapy, and CD4 count. PMID- 20009767 TI - Isometric handgrip exercise and resting blood pressure: a meta-analysis of randomized controlled trials. AB - OBJECTIVE: To examine the efficacy of isometric handgrip exercise for reducing resting SBP and DBP in adult humans. METHODS: Meta-analysis of studies retrieved from five electronic databases as well as cross-referencing from identified articles. The criteria for inclusion were randomized controlled trials published in any language over an approximate 38-year period (1 January 1971 to 1 February 2009), isometric handgrip training of at least 4 weeks performed by adults of at least 18 years of age, and data for changes in resting SBP and DBP available. Dual coding of studies was performed by both investigators. Data were analyzed a priori using random-effects models and nonparametric 95% bootstrap percentile confidence intervals (BCIs, 5000 iterations). Because of the small sample size, analyses were also performed using fixed-effects models post hoc. RESULTS: Eighty one men and women (42 exercise and 39 control) from three of 287 reviewed studies were pooled for analysis. Using random-effects models, statistically significant exercise minus control group reductions of approximately 10% were observed for both resting SBP and DBP (SBP: Xd , -13.4 mmHg; 95% BCI, -15.3 to -11.0 mmHg and DBP: X , -7.8 mmHg; 95% BCI, -16.5 to -3.0 mmHg). Results were also statistically significant when fixed-effects models were used (SBP: X , -13.8 mmHg; 95% BCI, 15.3 to -11.0 mmHg and DBP: X , -6.1 mmHg; 95% BCI, -16.5 to -3.2 mmHg). CONCLUSION: Isometric handgrip exercise is efficacious for reducing resting SBP and DBP in adult humans. However, the generalizability of these findings is limited given the small number of studies included. PMID- 20009768 TI - Two-dimensional, sex-specific autosomal linkage scan of the number of sodium pump sites. AB - OBJECTIVES: The sodium pump consists of the membrane-bound enzyme sodium/potassium-ATPase, which exchanges internal sodium ions for external potassium ions. Obesity, hypertension, and diabetes associate with the activity of the sodium pump, motivating gene discovery for sodium pump number. METHODS: Variance components linkage analysis was applied to the number of red blood cell sodium pump sites measured by ouabain-binding assays on 1375 members of 46 Utah pedigrees. Both one-dimensional (1D) and two-dimensional (2D) autosome-wide linkage analyses of pump number were performed on the combined sample as well as separately on the male and female subsets. RESULTS: Two significant 1D linkages were identified: on chromosome 1p13 in the combined sample [1D logarithm of odds (LOD) score = 3.76] and on chromosome 17p21 in the female subset (1D LOD score = 3.24). In addition, two significant 2D linkages were identified in the female subset: on chromosome 10q22 interacting with chromosome 18q11 (2D LOD score = 7.18) and on chromosome 13q21 interacting with chromosome 4q31 (2D LOD score = 6.05). Single-nucleotide polymorphism rs17376826 in neuropeptide Y receptor Y2, an obesity-associated gene and a candidate in the chromosome 4q31 linkage region, is associated with pump number (P = 0.046 in the combined sample and P = 0.042 in the female subset). CONCLUSION: Pump number is influenced by multiple genes, possibly including neuropeptide Y receptor Y2. PMID- 20009770 TI - Does the ratio of serum aldosterone to plasma renin activity predict the efficacy of diuretics in hypertension? Results of RENALDO. AB - OBJECTIVES: We hypothesized that the aldosterone: renin ratio (ARR) predicts the antihypertensive response to mineralocorticoid receptor antagonist, spironolactone (SPIRO), when compared with bendroflumethiazide (BFZ). METHODS: We conducted a randomized, crossover, trial on hypertensive patients with either high ARR (HARR defined as >750 and plasma aldosterone >250 pmol/l) or low ARR (LARR defined as <300 and plasma renin activity <10 ng/ml per h). Each group took SPIRO 50 mg once daily for 12 weeks and BFZ 2.5 mg once daily for 12 weeks in random order separated by 2-week washout. Patients with mean 24-h systolic ambulatory blood pressure (SABP) at least 140 mmHg were included. Primary endpoint was difference in SABP between SPIRO and BFZ in patients with HARR compared with those with LARR. RESULTS: One hundred and eleven patients (60 HARR and 51 LARR) completed the study. SABP at 12 weeks in the HARR group was 129.4 mmHg on SPIRO and 134.4 mmHg on BFZ [difference -5.01; 95% confidence interval (CI) -7.51, -2.52; P < 0.0002]. In the LARR group, SABP was 129.7 mmHg on SPIRO and 133.1 mmHg on BFZ [difference -3.43 (95% CI -6.18, -0.68) P < 0.01]. Difference between groups (HARR vs. LARR) was -1.58 mmHg (95% CI 5.25, -2.08; not significant, P = 0.394). In a secondary analysis of the overall study population of 111 patients, SABP reduction with SPIRO 50 mg was superior to BFZ 2.5 mg [SPIRO -14.8 mmHg, BFZ -10.5 mmHg, difference -4.29 mmHg (95% CI -6.12, -2.46)]. Results were similar for secondary endpoints. Plasma renin activity or aldosterone did not predict blood pressure response to SPIRO. Results were independent of concomitant angiotensin-converting enzyme inhibitor or angiotensin receptor blocker use. CONCLUSION: The ARR did not predict the blood pressure response to SPIRO. SPIRO 50 mg was significantly more effective than BFZ 2.5 mg in lowering SABP irrespective of baseline ARR, plasma renin activity or aldosterone. PMID- 20009769 TI - Human dopamine beta-hydroxylase (DBH) regulatory polymorphism that influences enzymatic activity, autonomic function, and blood pressure. AB - RATIONALE: Dopamine beta-hydroxylase (DBH) plays an essential role in catecholamine synthesis by converting dopamine into norepinephrine. Here we systematically investigated DBH polymorphisms associated with enzymatic activity as well as autonomic and blood pressure (BP)/disease phenotypes in vivo. METHODS AND RESULTS: Seventy genetic variants were discovered at the locus; across ethnicities, much of the promoter was spanned by a 5' haplotype block, with a larger block spanning the promoter in whites than blacks. DBH secretion was predicted by genetic variants in the DBH promoter, rather than the amino acid coding region. The C allele of common promoter variant C-970T increased plasma DBH activity, epinephrine excretion, the heritable change in BP during environmental stress in twin pairs, and also predicted higher basal BP in three independent populations. Mutagenesis and expression studies with isolated/transfected DBH promoter/luciferase reporters in chromaffin cells indicated that variant C-970T was functional. C-970T partially disrupted consensus transcriptional motifs for n-MYC and MEF-2, and this variant affected not only basal expression, but also the response to exogenous/co-transfected n MYC or MEF-2; during chromatin immunoprecipitation, these two endogenous factors interacted with the motif. CONCLUSIONS: These results suggest that common DBH promoter variant C-970T plays a role in the pathogenesis of human essential hypertension: common genetic variation in the DBH promoter region seems to initiate a cascade of biochemical and physiological changes eventuating in alterations of basal BP. These observations suggest new molecular strategies for probing the pathophysiology, risk, and rational treatment of systemic hypertension. PMID- 20009771 TI - Split-course palliative radiotherapy for advanced non-small cell lung cancer. AB - PURPOSE: Palliative chest radiotherapy (RT) for lung malignancies is effective in relieving serious chest symptoms from tumor bleeding or mass effect on major airways, vessels, and nerves. Albeit an important subject, there is a lack of consensus for an optimal palliative RT regimen. We report the outcomes of a split course palliative chest RT, a frequently used schema at our institution. METHODS AND MATERIALS: Records of 140 patients treated between 1995 and 2006 were reviewed. Treatment was prescribed to an initial 25 Gy in 10 fractions through anterior-posterior/posterior-anterior beam arrangements. After a 2-week rest period, patients were selected to receive an additional 10 Gy (anterior posterior/posterior-anterior) followed by off-cord beams to a final dose of 50 to 62.5 Gy. Symptom relief and toxicity during RT and after completion of RT were assessed from clinician notes and patient-reported symptom inventory forms. Second, the impact on survival was assessed. RESULTS: Symptomatic relief was observed in 52 to 84% of patients with durable palliation in 58%. There were no grade 3 to 5 toxicities. Grades 1 and 2 esophagitis and pneumonitis were observed in 34 and 8% patients, respectively. Median survival was 5 months. CONCLUSIONS: A majority of patients experienced symptomatic improvement. The built-in 2-week break allowed for selection of patients for high-dose palliative radiation and balanced treatment benefits with potential side effects. Cancer survival was not adversely affected by treatments in this population with mostly advanced disease. This regimen is a viable option for patients who cannot tolerate a protracted, uninterrupted course of treatment. PMID- 20009772 TI - Can a thoracic surgeon identify lymph node metastases during surgery based on their size? Analysis of 844 metastatic and 10,462 nonmetastatic lymph nodes. AB - OBJECTIVE: A metastatic lymph node commonly becomes enlarge; however, there is limited data available with regard to the direct measurement of lymph nodes and their clinicopathologic characteristics. METHODS: The size of dissected lymph node was quantified in a total of 848 nodes with metastasis and 10,462 nodes without metastasis from 454 patients with lung cancer who underwent a pulmonary resection with lymph node dissection. RESULTS: The short axis and the volume of the metastatic lymph nodes were significantly greater than those of the nonmetastatic ones. The smaller the lymph node, the less frequently the lymph nodes were metastatic; however, the ratios of nodes smaller than the fifth largest lymph node with metastasis of adenocarcinoma and squamous cell carcinoma were 21.8 to 26.2%, respectively. When the hilar and mediastinal lymph node stations were examined, 1.14 to 4.00% of the lung cancer patients had lymph node metastasis in small lymph node despite having no metastases in the largest and second largest lymph nodes. CONCLUSIONS: The small lymph nodes in the hilar or mediastinal stations frequently had metastases of carcinoma even though largest and second largest lymph nodes were negative for metastases, especially in adenocarcinoma cases. Surgical oncologists should, therefore, perform systemic lymph node dissection, and not sampling, during a pulmonary resection of lung cancer. PMID- 20009773 TI - Video-assisted thoracoscopic surgery (VATS) for patients with solitary fibrous tumors of the pleura. AB - OBJECTIVES: To present our experience of video-assisted thoracoscopic surgery (VATS) for patients with solitary fibrous tumors of the pleura (SFTPs) and to discuss the treatment of choice of such neoplasms. METHODS: Between June 2000 and September 2008, 21 patients with SFTPs (9 men and 12 women) underwent VATS at our department. The mean age was 52.5 years (range, 33-76 years). RESULTS: Surgical excision was performed in all patients. Surgical excision was performed by VATS in 15 patients (71.4%), by VATS plus a small thoracotomy (<5 cm) in 4 patients (19.0%), and by posterolateral thoracotomy accompanied by VATS in 2 patients (9.5%). Mean chest drain duration was 2.3 days (range, 1-4 days), and the mean hospital stay was 7.2 days (range, 4-15 days). There were 18 pathologically benign SFTP cases (85.7%) and 3 malignant SFTP cases (14.3%). There was no operative morbidity or mortality. No recurrence or metastasis of SFTPs developed during postoperative median follow-up period of 43 months. CONCLUSIONS: Complete resection and close follow-up for years after operation is recommended for SFTPs. VATS may play an important role in reducing the size of the thoracotomy incision in the treatment of SFTPs, which results in less invasive surgery. PMID- 20009774 TI - Impact of postoperative radiation after esophagectomy for esophageal cancer. AB - INTRODUCTION: Though postoperative radiation for esophageal cancer is offered in selected cases, there is conflicting evidence as to whether it improves overall survival (OS). We performed a retrospective analysis using the Surveillance Epidemiology and End Results database to analyze the impact of adjuvant radiation in a large cohort of patients. METHODS: From 1998 to 2005, patients diagnosed with stage T3-4N0M0 or T1-4N1M0 esophageal adenocarcinoma (AC) or squamous cell carcinoma (SCC) who were definitively treated with esophagectomy, with or without postoperative radiation, were selected. Kaplan-Meier and Cox regression analysis were used to compare OS and disease-specific survival (DSS). RESULTS: A total of 1046 patients met the selection criteria: 683 (65.3%) received surgery alone and 363 (34.7%) received postoperative radiation. For American Joint Committee on Cancer stage III esophageal carcinoma (T3N1M0 or T4N0-1M0), there was significant improvement in median and 3-year OS (p < 0.001) and DSS (p < 0.001), respectively. This benefit was present for both SCC and AC. However, for American Joint Committee on Cancer stages IIA and IIB disease there was no significant differences in OS or DSS. Multivariate analysis revealed that postoperative radiation was the most significant predictor for improved OS (hazard ratio 0.70, 95% confidence interval 0.59-0.83, p < 0.001). CONCLUSIONS: This large population based review supports the use of postoperative radiation for stage III SCC and AC of the esophagus. Given the retrospective nature of this study, until appropriately powered randomized trials confirm these results, caution should be used before broadly applying these findings in clinical practice. PMID- 20009775 TI - A phase II study of perioperative concurrent chemotherapy, gefitinib, and hyperfractionated radiation followed by maintenance gefitinib in locoregionally advanced esophagus and gastroesophageal junction cancer. AB - BACKGROUND: Concurrent chemoradiotherapy (CCRT) for locoregionally advanced esophageal or gastroesophageal junction cancer produces high locoregional control rates but suboptimal distant metastatic control (DMC) and overall survival. This phase II study added gefitinib (G) to our previously tested CCRT regimen in an effort to improve these outcomes. METHODS: Eligibility required T3, N1, or M1a esophageal or gastroesophageal junction squamous cell or adenocarcinoma staged by esophageal ultrasound and positron emission tomography/computed tomography. Four day continuous intravenous infusions of cisplatin (20 mg/m/d) and fluorouracil (1000 mg/m/d) began on day 1 of preoperative radiation (30 Gy and 1.5 Gy bid). Surgery followed in 4 to 6 weeks, and an identical course of CCRT 6 to 10 weeks postoperatively. G 250 mg/d was given with preoperative CCRT for 4 weeks and restarted with postoperative therapy for 2 years. Results were retrospectively compared with our historical series of 93 patients given CCRT without G. RESULTS: Between April 2003 and July 2006, 80 patients were enrolled. Patient and tumor characteristics were similar to our historical series. G did not increase toxicity except for development of rash in 42 (53%) and diarrhea in 44 (55%) 3 year Kaplan-Meier estimates (G versus non-G treated patients) included: overall survival (42% versus 28%, p = 0.06), DMC (40% versus 32%, p = 0.33), and locoregional control (76% versus 77%, p = 0.74). Intolerance for G maintenance occurred in 48% of patients. Patients who experienced G related diarrhea appeared to have improved outcomes. CONCLUSIONS: Although G did not worsen CCRT toxicity, maintenance therapy proved difficult. This contemporary cohort of patients enjoyed superior survival, which does not solely reflect a decrease in DMC and merits further investigation. PMID- 20009776 TI - Resuscitation with hydroxyethyl starch solution prevents bone marrow mononuclear apoptosis in a rat trauma-hemorrhagic shock model. AB - BACKGROUND: : Trauma-hemorrhagic shock (T/HS) has been associated with multiorgan dysfunction, including bone marrow failure. This study examined apoptosis and morphologic alterations in bone marrow mononuclear cells (BMMNCs) with different volume therapies after T/HS. METHODS: : T/HS was induced in groups of male Sprague-Dawley rats through a fracture of the left femur and continual bleeding for 30 minutes, followed by resuscitation with Ringer's lactate solution (RL), 6% hydroxyethyl starch solution (HES), or 5% albumin (ALB). Mean arterial blood pressure was monitored during the T/HS and resuscitation, and the impacts of various resuscitative fluids on apoptosis and morphology of BMMNCs at 24 hours and 48 hours after resuscitation were examined using flow cytometry, transferase mediated dUTP nick-end labeling assay, and hematoxylin and eosin staining. RESULTS: : Fluctuations in mean arterial blood pressure were homogenous among the three treatment groups. The percentage of early BMMNC apoptosis increased significantly at 24 hours and 48 hours (24.65% +/- 5.41% and 29.09% +/- 2.07%, respectively; p < 0.05), and the percentage of late BMMNC apoptosis increased to 13.43% +/- 2.82% (p < 0.05) at 48 hours in the T/HS + RL group. In contrast, resuscitation with HES alone dramatically attenuated the apoptosis. Resuscitation with ALB alleviated BMMNC apoptosis, except for late apoptosis at 48 hours. A greater number of apoptotic BMMNCs as well as morphologic alterations were shown using the transferase-mediated dUTP nick-end labeling assay and hematoxylin and eosin stain in the T/HS + RL group than in the HES or ALB groups. CONCLUSION: : Intravascular volume replacement with HES showed prevention of BMMNC apoptosis at first 48 hours after T/HS compared with RL and ALB. These findings provide new insights into the intervention mechanism of HES on T/HS-related multiorgan dysfunction. PMID- 20009777 TI - Quality variation and its impact on costs and satisfaction: evidence from the QIDS study. AB - BACKGROUND: Improving the quality of inpatient hospital care is increasingly attainable in a variety of settings. However, the relationship between rising quality and costs is unclear; similarly the relationship between varying levels of quality and a patient's satisfaction remains poorly defined. METHODS: We use data from the Quality Improvement Demonstration Study (QIDS) based in 30 district hospitals in the Philippines. There were 974 children in the study; these children were cared for by 43 physicians. To measure quality of care, the physicians completed vignettes, a valid and inexpensive measure. Patient exit surveys were given to parents of children on the day of discharge, collecting information on services and hospital charges for the inpatient stay, payment sources for the hospitalization, and the Patient Satisfaction Survey (PSQ-18). RESULTS: We found a nonlinear relationship between quality and hospital charges: at low levels of quality improvements are linked to lower hospital charges. However, as quality improves further, these changes lead to higher charges. Higher quality also demonstrated a similar nonlinear relationship with patient satisfaction. CONCLUSIONS: The U-shaped association between quality and hospital charges suggests that targeting the lowest quality providers may decrease costs. The similar relationship between patient-reported satisfaction and quality improvement suggests that investments in quality will raise satisfaction, perhaps even when charges are increased. PMID- 20009778 TI - Characteristics of insured patients with persistent gaps in diabetes care services: the Translating Research into Action for Diabetes (TRIAD) study. AB - BACKGROUND: Although preventing diabetes complications requires long-term management, little is known about which patients persistently fail to get recommended care. OBJECTIVE: To determine the frequency and correlates of persistent, long-term gaps in diabetes care. METHOD: : The study population included 8392 patients with diabetes. Patient surveys and medical records from 10 health plans over 3 years provided data on socioeconomic characteristics, access to care, social support, and mental and physical health, and diabetes preventive care services. We defined a "persistent gap" as a participant's missing a preventive care service for the entire 3 years. Services considered included hemoglobin A1c, cholesterol, and albuminuria tests, and foot and dilated eye examinations. RESULTS: Thirty percent of participants had at least 1 persistent gap. The most common gaps were lipid testing (11.6%), microalbuminuria testing (9.7%), and eye examinations (9.0%). Persistent gaps were 18% to 42% higher for young patients, lean persons, those with low income, employed persons, smokers, those with diabetes less than 5 years, and patients with none or 1 comorbid conditions. Sex, education, marital status, family demands, transportation, trust in physicians, and mental health were not associated with gaps in care. CONCLUSIONS: Persistent gaps in diabetes care are common even among insured patients. Patients with lower income, younger age, fewer years of diabetes, having fewer comorbidities, taking fewer medications, and poor health behaviors are vulnerable to persistent gaps in care and a group who warrant targeted interventions to improve preventive diabetes care. PMID- 20009779 TI - The management of peripheral facial nerve palsy: "paresis" versus "paralysis" and sources of ambiguity in study designs. AB - OBJECTIVE: Conservative management of idiopathic or herpetic acute peripheral facial palsy (herpes zoster oticus, HZO) often leads to a favorable outcome. However, recent multicenter studies have challenged the necessity of antivirals. Whereas large numbers of patients are required to reveal statistical differences in a disease with an overall positive outcome, surprisingly few studies differentiate between patients with paresis and paralysis. Analyzing our own prospective cohort of patients and reviewing the current literature on conservative treatment of Bell's palsy and HZO, we reveal the importance of initial baseline assessment of the disease course to predict the outcome and to validate the impact of medical treatment options. STUDY DESIGN AND DATA SOURCE: Prospective analysis of consecutive patients referred to 2 tertiary referral centers and research on the Cochrane Library for current updates of their previous reviews and search of MEDLINE (1976-2009) for randomized trials on conservative treatment of acute facial palsy were conducted. METHODS: One hundred ninety-six patients with Bell's palsy or HZO were followed up prospectively until complete recovery or at least for 12 months. The numeric Fisch score (FS) was used to classify facial function, and patients were separated between incomplete palsy (=paresis) and complete paralysis. Electroneuronography (ENoG) was used to further subdivide patients with paralysis. The treatment protocol was independent of the ongoing investigation including prednisone and valacyclovir in most patients. A total of 250 previous studies on facial palsy outcome were evaluated regarding their distinction between different severity scores at baseline and its impact on treatment outcome. Trials not making the distinction between paresis and paralysis at baseline and with an insufficient follow-up of less than 12 months were excluded. RESULTS: In the Bell's and HZO paresis group, all except 1 patient recovered completely, most of them within 3 months, independent of the treatment regimen. In the Bell's paralysis group, 38 patients (70%) recovered completely after 1 year, including 94% of patients with a denervation by ENoG of less than 90%. Thirty percent of Bell's paralysis patients recovered incompletely, revealing the worst outcome in patients with a 100% denervation on ENoG. None of the 4 patients with HZO and ENoG denervation of more than 90% recovered to normal facial function. We found a highly significant difference regarding the time course and final outcome in patients with incomplete palsies versus total paralysis; however, only 3 of 250 studies make this distinction. CONCLUSION: The time course for improvement and the extent of recovery is significantly different in patients presenting with an incomplete facial nerve paresis compared with patients with a total paralysis. Whereas the term "palsy" includes both entities, the term "paralysis" should only be used to describe total loss of nerve function. Patients with incomplete acute Bell's palsy (paresis) should start to improve their facial function early (1-2 wk after onset) and are expected to recover completely within 3 months. These patients do not benefit from antiviral medications and most likely do not profit from systemic steroids. Mixing patients with different severity of palsies will always lead to controversial results. PMID- 20009780 TI - Corticosteroids in the treatment of vestibular neuritis: a systematic review and meta-analysis. AB - OBJECTIVE: To systematically review and meta-analyze the results of all randomized controlled trials comparing corticosteroids with placebo for the treatment of patients with vestibular neuritis. DATA SOURCES: An electronic search was performed in MEDLINE, EMBASE, Cochrane Library, and CENTRAL databases, and then extensive hand-searching was performed for the identification of relevant studies. No time and language limitations were applied. STUDY SELECTION: Prospective randomized controlled trials comparing corticosteroids with placebo for the treatment of patients with vestibular neuritis. DATA EXTRACTION: Odds ratios (ORs), weighted mean differences (WMD), 95% confidence intervals (CIs), and tests for heterogeneity were reported. DATA SYNTHESIS: Four studies were eventually identified and systematically reviewed. Meta-analysis was feasible for 3 studies. Regarding the recovery of clinical symptoms, the proportion of patients with clinical recovery at 1 month after the initiation of therapy did not differ significantly between the corticosteroids and placebo groups (OR, 1.45; 95% CI, 0.26-8.01; p = 0.67). The proportion of patients with caloric complete recovery was significantly different between the corticosteroids and placebo groups both at 1 (OR, 12.64; 95% CI, 2.6-61.52; p = 0.002; heterogeneity, p = 0.53; fixed effects model) and 12 months (OR, 3.35; 95% CI, 1.45-7.76; p = 0.005; heterogeneity, p = 0.03; random effects model) after the initiation of therapy. The caloric extent of canal paresis at 12 months after the initiation of therapy seemed to differ significantly between patients who received corticosteroids and those who received placebo (WMD, -12.15; 95% CI, -19.85 to 4.46; p < 0.05; heterogeneity, p < 0.05; random effects model). CONCLUSION: The present systematic review and meta-analysis, based on the currently available evidence, suggests that corticosteroids improve only the caloric extent and recovery of canal paresis of patients with vestibular neuritis. At present, clinical recovery does not seem be better in patients receiving corticosteroids. PMID- 20009781 TI - Rehabilitation of canal wall down mastoidectomy using a titanium ear canal implant. AB - OBJECTIVE: To describe the rehabilitation of canal wall down mastoidectomy cavities using a custom-made titanium wall implant and to analyze the benefits and the long-term functional results. STUDY DESIGN: Retrospective analysis. SETTING: Tertiary care center. PATIENTS AND METHODS: We have reviewed the data of 113 consecutive operated ears from 106 patients during a 9-year period (1993 2002). The following preoperative and postoperative criteria were compared: the persistence or resolution of otorrhea, cholesteatoma, and vertigo; external auditory canal healing, position of the titanium prosthesis, and tympanic membrane findings. In addition, the hearing status was evaluated by comparing the preoperative data with the 6-month and last follow-up postoperative data to deduce the air-bone gap (ABG) and the auditory gains. RESULTS: Operated ears (88.5%) were dry after the initial rehabilitation, and 97.3% were dry after a revision surgery for defective canal healing. Most patients (96.5%) were free of vertigo. The external auditory canal wall was in a good position in all cases and allowed for a primary skin healing process in 84% of cases; the remaining patients underwent revision surgery under local anesthesia to complete the healing. The reconstructed drum was stable and anatomic in 73% of the cases (n = 83) after the first surgery. Thirty-six patients (32%) underwent various revision procedures: aeration tube placement for 16 patients with mesotympanic retraction and/or serous effusion, revision surgery for 12 patients with attic retraction or evidence of cholesteatoma, and revision for 5 patients with perforation. At last follow-up, 100% of the ear canals were healed, and 85% of patients had a normal tympanic membrane. No residual cholesteatoma was found at last follow-up. Thirty six percent of patients had an ABG within 20 dB at last follow-up, and 69% within 30 dB. CONCLUSION: The rehabilitation of canal wall down mastoidectomies improves the quality of life of patients regarding cholesteatoma recurrence, otorrhea, and vertigo in more than 95% of cases and stabilizes the hearing loss to less than 30 dB of ABG for more than two thirds of patients. The main unresolved problem is the persistent dysfunction of the middle ear cavity, with unsatisfactory auditory improvements. Middle ear implants represent in selected cases a new solution to rehabilitate the refractory hearing losses. PMID- 20009782 TI - Blast exposure: vestibular consequences and associated characteristics. AB - OBJECTIVE: To characterize vestibular and related symptoms seen after blast exposure. STUDY DESIGN: Prospective single-subject design. SETTINGS: Tertiary care facility and military field hospital. PATIENTS: Active-duty US Military personnel exposed to blast(s) in Iraq and/or Afghanistan. INTERVENTIONS: Vestibular function tests, auditory tests, and a structured history and physical examination. RESULTS: Blast exposure induced vestibular disorders, and related symptoms are significantly different than those seen in blunt head trauma. The vestibular characteristics and objective tests of vestibular function significantly worsen in blast-exposed patients as a function of time between injury and presentation. CONCLUSION: Blast exposure produces a unique set of vestibular disorders and associated symptoms that progress over time. Understanding the characteristics of these symptoms as they vary over time may be critical in designing treatment strategies that modify long-term outcome. PMID- 20009783 TI - The suprameatal approach: a safe alternative surgical technique for cochlear implantation. AB - OBJECTIVE: To report on surgical complications arising postoperatively in 104 patients undergoing cochlear implantation surgery using the suprameatal approach (SMA). Second, to examine the advantages and disadvantages of the SMA technique compared with the classic mastoidectomy using the posterior tympanotomy approach. STUDY DESIGN: Retrospective study assessing surgical complications in deaf adults and children undergoing cochlear implantation. SETTING: Tertiary referral center for cochlear implantation (Academic Medical Centre). PATIENTS: The mean age at the time of surgery was 39.6 years (1.0-82.3 yr), and the mean duration of deafness was 26.3 years (0.3-66.0 yr). The main cause was a congenital hearing loss (30.8%) and a progressive sensorineural hearing loss e.c.i. (25.0%), followed by meningitis (12.5%) and otosclerosis (6.7%). The mean duration of follow-up after surgery was 25.7 months (range, 3.0-59.0 mo). RESULTS: The overall major complication rate was 3.7% (4 of 107). All complications developed postoperatively. The major complications consisted of extrusion of the implant due to wound infection (n = 2), a wrong route for the electrode (n = 1), and device failure (n = 1). The minor complication rate was 23.4% (25 of 107). The mean time taken for cochlear implantation was 111.7 minutes (range, 60.0-261.1 min). CONCLUSION: This study confirms the SMA is a safe, simple, and quick technique that is feasible for cochlear implantation in most cases. Although the stretching of the electrode array when it enters the scala tympani and a low lying dura could present a potential restriction for the SMA technique, our results do not support this hypothesis. PMID- 20009784 TI - Pediatric infectious diseases: 2009 update for the Rogers' Textbook of Pediatric Intensive Care. AB - OBJECTIVE: To review important articles in the field of infectious diseases that pertain to the care of children in pediatric critical care units, published subsequent to the fourth edition of the Rogers' Textbook of Pediatric Intensive Care. DATA SOURCES: The U.S. National Library of Medicine was searched for the terms: critical care, nosocomial, antimicrobial resistance, opportunistic infection, sepsis, central nervous system infections, encephalitis, meningitis bacterial, meningitis-tuberculous, brain abscess, measles, dengue, hemorrhagic fever, human immunodeficiency virus infection (HIV), opportunistic infections, fungal infections, tetanus, diphtheria, botulism, toxic shock syndrome, and pediatrics. STUDY SELECTION AND DATA EXTRACTION: Promising articles were reviewed and the decision to include them or not in this review was made by the authors, based on clinical relevance. DATA SYNTHESIS: Articles were included based on their relevance to specific chapters included in this section of the textbook. CONCLUSIONS: Significant advances continue to be made in our understanding of specific diseases as well as the approach to treatment. There are significant variations in outcome from specific infectious diseases in developing countries compared with the developed world. The looming problem of antimicrobial resistance and relative lack of new anti-infective agents in development is an issue that will be faced by pediatric intensive care units throughout the world in the near future. Updated evidence-based guidelines have appeared for early treatment of septic shock in children, and on prevention and treatment of opportunistic infections in adults and adolescents with human immunodeficiency virus. In patients with measles, use of oral co-trimoxazole or amoxicillin reduces the risk of secondary bacterial infections of respiratory tract; however, the same may not be true for other systemic viral infections, such as influenza. In patients with acute bacterial meningitis, maintenance fluids-instead of restricted fluids-and use of glycerol may improve the outcomes; however, the role of dexamethasone in prevention of adverse outcome needs reevaluation. Intravenous use of botulism immune globulin decreases the length of hospital stay and mechanical ventilation. Pediatric patients with aspergillosis have radiologic findings distinct from those of adults and present more often with nodules, and only rarely with cavitation; early focal surgical resection may improve the survival of these patients. PMID- 20009786 TI - Multimethod adherence assessment in children with perinatally acquired HIV-1: the influence of off-schedule dosing in predicting biological markers. AB - To improve upon adherence assessment in children with HIV, multimethod adherence strategies (pill count, missed doses, off-schedule dosing) were conducted concurrent with viral load and CD4% biomarker assays. Off-schedule dosing predicted both health status markers, while the more common strategies did not. Findings support inclusion of off-schedule dosing concurrent with collection of biomarkers to assess adherence in children with HIV. PMID- 20009785 TI - Leukocyte subset-derived genomewide expression profiles in pediatric septic shock. AB - OBJECTIVE: To directly assess whether genomewide expression profiles derived from leukocyte subsets are comparable to that of whole blood as measured by enrichment for genes corresponding to metabolic and signaling pathways. DESIGN: Prospective observational study involving microarray-based bioinformatics based on RNA individually derived from whole blood, neutrophils, monocytes, and lymphocytes, respectively. SETTING: Three pediatric intensive care units in the United States. PATIENTS: Children < or =10 yrs of age: five normal control subjects and 13 meeting criteria for septic shock on day 1 of presentation to the pediatric intensive care unit. INTERVENTIONS: None other than standard care. MEASUREMENTS AND MAIN RESULTS: Baseline analyses using whole blood-derived RNA demonstrated increased expression of genes corresponding to signaling pathways involving innate immunity, redox balance, and protein ubiquitination and decreased expression of genes corresponding to the adaptive immune system. Subsequent analyses using leukocyte-specific RNA were congruent with the gene expression profiles demonstrated using whole blood-derived RNA as measured by enrichment for genes corresponding to metabolic and signaling pathways. Gene network analysis, derived from a composite gene list involving the individual gene expression profiles of neutrophils, monocytes, and lymphocytes, respectively, revealed a gene network corresponding to antigen presentation, cell-mediated immunity, and humoral-mediated immunity. Finally, a subanalysis focused on network gene nodes localized to the nuclear compartment revealed functional annotations related to transcriptional repression and epigenetic regulation. CONCLUSIONS: These data demonstrate that genome-level repression of adaptive immunity gene programs early in the course of pediatric septic shock remained evident when analyses were conducted using leukocyte subset-specific RNA. PMID- 20009787 TI - Construction and clinical application of a human tissue-engineered epidermal membrane. AB - BACKGROUND: Prolonged healing times and hypertrophic scarring of the donor site for split-thickness-skin grafts thicker than 0.3 mm are common problems that continue to challenge plastic surgeons in the clinic. As such, a human tissue engineered epidermal membrane was constructed to promote wound healing and reduce scar hypertrophy. METHODS: An artificial allogenic epidermis was created in vitro using human keratinocytes and chitosan-gelatin membrane. Split-thickness skin graft donor sites were divided into three treatment groups: those covered with the combined keratinocyte/chitosan-gelatin membrane, those covered with chitosan gelatin membrane only (control group), and those covered with traditional petroleum jelly gauze (blank group). The degree of wound healing was assessed at various time points after the operation by gross observation, hematoxylin and eosin staining, immunohistochemistry, and an assay of type I collagen using the picrosirius polarization method. Reverse-transcriptase polymerase chain reaction detection of the Y chromosome was also performed to distinguish between sexes. RESULTS: Over a 6-month observation period, treatment with the human tissue engineered epidermal membrane (keratinocyte/chitosan-gelatin) appeared to decrease donor-site healing time (48 wounds in 24 cases). Average healing time was 8.1 +/- 1.3 days for the keratinocyte/chitosan-gelatin group, 16.4 +/- 1.7 days for the chitosan-gelatin group, and 22.9 +/- 4.2 days for the blank group. The artificial epidermis survived well and maintained a normal structure. Furthermore, hypertrophic scar formation was less severe for these wounds. CONCLUSIONS: Keratinocyte/chitosan-gelatin membranes can be constructed in vitro and survive temporarily in vivo. They can be used to promote wound healing and reduce the severity of hypertrophic scarring of skin graft donor sites. PMID- 20009788 TI - Fascial flap reconstruction of the hand: a single surgeon's 30-year experience. AB - BACKGROUND: The reconstruction of complex hand wounds is challenging due to the requirements for thin and pliable coverage with a reliable vascular supply, potential for sensibility, and provision of a gliding surface. Fascial flaps represent an excellent option for the reconstruction of these complicated defects. METHODS: A retrospective review of fascial flap reconstructive procedures to the hand undertaken by a single microsurgeon was performed for operations occurring between 1979 and 2009. Both pedicled and free tissue transfer procedures were included in both pediatric and adult patients. Data were culled from a combination of patient charts, hospital records, radiographic studies, and clinical photographs. RESULTS: Sixty fascial flap reconstructive procedures to the hand were analyzed in 60 patients from the defined 30-year period. The most common pathological process necessitating reconstruction was acute trauma (n = 32, 53 percent). Most of the soft-tissue injuries included in the study sample were located on the dorsal hand and wrist (n = 27, 45 percent). The most commonly utilized reconstructive modality was the temporoparietal fascial flap (n = 35, 58 percent). Most reconstructions were completed as free tissue transfers (n = 46, 77 percent). Perioperative complications were relatively minor; no flap losses were recorded. All cases studied demonstrated excellent long-term coverage with no evidence of underlying tendon adhesion or contracture. CONCLUSION: Fascial flaps represent an excellent option for coverage of soft-tissue defects of the hand that are not amenable to reconstruction with skin grafting alone, particularly for localized defects with denuded tendons or exposed joints. PMID- 20009789 TI - Longitudinal outcomes and application of the subunit principle to 165 foot and ankle free tissue transfers. AB - BACKGROUND: Free tissue transfer to the lower extremity has become a well established reconstructive modality. The purpose of this study was to develop a "subunit" approach to patients undergoing free tissue transfer for foot and ankle wounds to help further define subunit-specific functional and aesthetic operative goals. METHODS: The institutional review board approved this retrospective review of 161 patients who underwent free tissue transplantation for foot and ankle wounds between March 1, 1997, and February 28, 2007, at a single institution. Endpoints included flap-related complications, secondary surgery, time to ambulation, flap stability, and limb salvage. RESULTS: The most common types of wounds treated were trauma-related [n = 120 (75 percent)], diabetes-related [n = 24 (15 percent)], and oncologic defects [n = 8 (5 percent)]. Ten different donor sites were used for reconstruction, with the latissimus dorsi flap being the most common. The mean follow-up time was 26.9 months (range, 0.5 to 130 months). Mean time to ambulation was 3.1 months (range, 0.75 to 14 months). Overall, 11 percent of patients required revision surgery for flap instability at a mean time of 25.3 months after flap surgery. Wounds located over the heel (subunit 5) were most likely to develop instability (Fisher's exact test, p < 0.05). The overall 5-year limb salvage rate as determined by Kaplan-Meier analysis was 89 percent. CONCLUSIONS: The use of free tissue transplantation for treatment of foot and ankle wounds is associated with a high rate of limb salvage. Although a variety of flaps may be used, the application of the subunit principle can assist surgeons in designing flaps that will address subunit-specific functional and aesthetic concerns. PMID- 20009790 TI - Preoperative sizing in breast augmentation. AB - BACKGROUND: Implant size selection in breast augmentation patients is one of many variables to be determined before surgery. Few methods exist today that allow the patient to participate in this process and accurately determine optimal size. The authors describe a simple method of preoperative sizing using silicone implant samples. METHODS: A total of 567 patients underwent breast augmentation: 297 had surgery before implementation of preoperative sizing and 270 patients were sized preoperatively. Sizing consisted of fitting the patients with various size silicone implants in a larger bra at least twice before surgery to determine desired size. Surveys were sent to both groups to inquire about overall satisfaction, how many preferred a different size postoperatively, and how many ultimately underwent size change surgery. RESULTS: One hundred two responses (34.3 percent) were obtained from the control group and 142 (52.6 percent) were obtained from the sized group. Sized patients received smaller implants (average, 276.6 cc nonsized versus 246.4 cc sized; p < 0.001). Four patients (1.4 percent) in the control group underwent a size change procedure compared with none in the sized group. In the sized cohort, 69 percent believe they are the size that the process predicted, 21 percent are smaller, 9 percent are larger, and 1 percent did not answer the question. CONCLUSIONS: Sized patients were more satisfied than controls and fewer were interested in having a different size implant postoperatively. Sized patients indicated that preoperative sizing was both helpful and reasonably accurate in predicting final breast size. PMID- 20009791 TI - Broad application of the endoscope for orbital floor reconstruction: long-term follow-up results. AB - BACKGROUND: Transantral endoscopic repair of orbital floor fracture minimizes the risk of implant misplacement and avoids complications associated with traditional lower eyelid approaches. Contrary to most publications, its application is not limited to early intervention of pure orbital blow-out fractures. The authors present their results of broad application of this technique, with longer-term patients' follow-up. METHODS: A retrospective review of 32 patients over a 10 year period (March of 1998 to June of 2008) was performed. The mean duration of follow-up was 27.5 months (range, 4 months to 10 years). Sixteen patients (50 percent) had associated zygoma, inferior orbital rim, and Le Fort I fractures. Twenty-five patients (78.1 percent) had enophthalmos, with 14 of these cases being 2 mm or more in severity. Diplopia was present in 15 patients (46.9 percent) preoperatively. Operations were performed within 2 weeks for 25 patients (78.1 percent). Twenty-eight patients required orbital floor reconstructions. Four of five patients with associated orbital medial wall fractures underwent simultaneous orbital medial wall reconstruction. RESULTS: All patients had successful reduction of orbital fractures. Two patients had residual enophthalmos of 1 mm postoperatively. Due to delay in surgery and the nature of injury, only 11 of the 15 patients with diplopia had complete resolution after surgery. One patient required drainage of recurrent sinusitis 1 year after surgery. CONCLUSIONS: In suitably selected patients with orbital floor fractures, the transantral endoscopic approach is safe and reliable. Delayed surgery or associated zygomaticomaxillary complex fractures are not contraindications for the use of this technique. Long-term follow-up showed maintenance of the surgical results. PMID- 20009792 TI - Transgene expression in a model of composite tissue allotransplantation. AB - BACKGROUND: Composite tissue allografting may be an ideal solution to many problems requiring reconstructive surgery. Unfortunately, complications associated with chronic immunocompromise are major impediments to widespread use of composite tissue allografting. Current immunosuppressive and immunomodulatory paradigms focus on modification of the recipient through global immunosuppression or donor/recipient chimerism. Alternatively, modifying the allograft to block rejection or promote tolerance could confine deleterious immunosuppressive effects to the graft or eliminate graft rejection. However, a technique introducing genetic information into the transplant is needed. The authors demonstrate the first model for expressing a gene of interest locally in a hind limb transplant. METHODS: Using a rat hind-limb transplant model, the authors tested the ability of naked DNA infusion, cationic polymer/DNA complex transfection, and adenoviral vector transduction to introduce genetic material into the composite tissue allograft. The marker genes luciferase and green fluorescent protein were used to follow gene expression. RESULTS: Recombinant adenovirus showed rapid, high-level expression of marker genes in the graft, with no detectable expression in recipient animals. Expression was detectable at 18 hours and peaked at 7 days. Levels of expression were lower but above baseline at 4 weeks. CONCLUSIONS: Using an adenoviral vector system, the authors have selectively introduced a marker gene (luciferase) into a transplanted hind-limb rat model. Expression was rapid and seen in a variety of cell types. Adenovirus infection had no impact on limb rejection. This method may be a powerful tool for genetically modifying composite tissue allografts and may contribute to immune tolerance and more widespread use of composite tissue allograft surgery. PMID- 20009793 TI - Appearance is a function of the face. AB - BACKGROUND: Increasingly, third-party insurers deny coverage to patients with posttraumatic and congenital facial deformities because these are not seen as "functional." Recent facial transplants have demonstrated that severely deformed patients are willing to undergo potentially life-threatening surgery in search of a normal physiognomy. Scant quantitative research exists that objectively documents appearance as a primary "function" of the face. This study was designed to establish a population-based definition of the functions of the human face, rank importance of the face among various anatomical areas, and determine the risk value the average person places on a normal appearance. METHODS: Voluntary adult subjects (n = 210) in three states aged 18 to 75 years were recruited using a quota sampling technique. Subjects completed study questionnaires of demography and bias using the Gamble Chance of Death Questionnaire and the Rosenberg Self Esteem Scale. RESULTS: The face ranked as the most important anatomical area for functional reconstruction. Appearance was the fifth most important function of the face, after breathing, sight, speech, and eating. Normal facial appearance was rated as very important for one to be a functioning member of American society (p = 0.01) by 49 percent. One in seven subjects (13 percent) would accept a 30 to 45 percent risk of death to obtain a "normal" face. CONCLUSIONS: Normal appearance is a primary function of the face, based on a large, culturally diverse population sample across the lifespan. Normal appearance ranks above smell and expression as a function. Restoration of facial appearance is ranked the most important anatomical area for repair. Normal facial appearance is very important for one to be a functional member of American society. PMID- 20009794 TI - Capsular contracture and possible implant rupture: is magnetic resonance imaging useful? AB - BACKGROUND: Currently, magnetic resonance imaging is considered the accepted standard to evaluate breast implant integrity. METHODS: To evaluate its utility in diagnosing ruptured silicone implants in the setting of capsular contracture and to correlate the preoperative assessment of implant integrity with or without magnetic resonance imaging with operative findings, 319 capsulectomies (171 patients with capsular contractures) were retrospectively reviewed. Preoperative magnetic resonance imaging was done on 160 implants, whereas the remaining 159 were evaluated using only physical examination and/or mammography. Postoperative results were analyzed to determine the sensitivity, specificity, and accuracy of preoperative magnetic resonance imaging in comparison with clinical and/or mammography evaluation alone. RESULTS: Although occasionally valuable, overall, preoperative magnetic resonance imaging was no more accurate than clinical evaluation with or without mammography in predicting implant status: magnetic resonance imaging 124 of 160 (78 percent) and clinical 121 of 159 (76 percent; p = 0.77). CONCLUSIONS: In the setting of capsular contracture, physical examination with or without mammogram is as accurate as magnetic resonance imaging in determining implant integrity. Although magnetic resonance imaging is a sensitive diagnostic tool, in symptomatic patients with capsular contracture, it cannot be viewed as infallible. PMID- 20009795 TI - Patient satisfaction and health-related quality of life following breast reconstruction: patient-reported outcomes among saline and silicone implant recipients. AB - BACKGROUND: In recent years, there has been a growing acceptance of the value of breast reconstruction. The majority of women who choose to proceed will undergo alloplastic reconstruction. The primary objective of this study was to determine whether the type of implant used in alloplastic breast reconstruction has an impact on patient-reported satisfaction and quality of life. METHODS: Patients were deemed eligible if they had completed alloplastic reconstruction at least 1 year before study initiation. Patients were contacted by mail: two questionnaires [the BREAST-Q and the European Organization for Research and Treatment of Cancer Quality of Life Questionnaire C30 (Br23) (EORTC QLQC30 (Br23))], a contact letter, and an incentive gift card were included. Scores were compared between silicone and saline implant recipients. RESULTS: Seventy-five silicone implant recipients and 68 saline implant recipients responded, for a response rate of 58 percent. BREAST-Q responses showed silicone implant recipients to have higher scores on all nine subscales. This difference reached statistical significance on four of nine subscales: overall satisfaction (p = 0.008), psychological well being (p = 0.032), sexual well-being (p = 0.05), and satisfaction with surgeon (p = 0.019). Regression analysis adjusted for follow-up time, timing of surgery, unilateral versus bilateral surgery, radiation, and age. Results from the EORTC QLQC30 (Br23) showed a statistically significant difference on two of 22 subscales: silicone recipients had higher overall physical function, and saline recipients had higher systemic side effects. CONCLUSIONS: This study has shown higher satisfaction with breast reconstruction in silicone gel implant recipients compared with saline recipients using the BREAST-Q. There was no difference in overall global health status between the two patient groups as measured by the EORTC-QLQC30 (Br23). PMID- 20009796 TI - Pharmacodynamic monitoring of calcineurin inhibition therapy: principles, performance, and perspectives. AB - The calcineurin inhibitors (CNIs) cyclosporin A and tacrolimus are immunosuppressive drugs used extensively in allograft recipients. These drugs show large interindividual pharmacokinetic variation and are associated with severe adverse affects, including nephrotoxicity and cardiovascular disease. In current practice, CNIs are combined with other immunosuppressive drugs such as steroids and mycophenolate mofetil. Dosage is titrated based on blood concentration measurement. For further optimization of calcineurin (CN) inhibition therapy, new monitoring strategies are required. Pharmacodynamic monitoring strategies constitute novel approaches for optimization of CNIs therapy. This review focuses on the general aspects of immunosuppressive drug pharmacodynamic monitoring and describes the methodologies used for monitoring CN inhibition therapy. Two different types of pharmacodynamic-monitoring strategies can be distinguished: (1) enzymatic strategies, which monitor inhibition of drug target enzyme activity, and (2) immunologic strategies, which measure cellular responsiveness after in vitro simulated immunologic responses. Enzymatic tests are drug type-specific markers in which CN activity is directly determined. Immunologic strategies measure immune responsiveness at several levels, such as mRNA transcripts (intracellular) concentrations/excretion of cytokines, expression of surface activation markers, and cell proliferation. This review also discusses analytical issues and clinical experience with these techniques. The call for new methodologies to evaluate immunosuppressive therapy has led to the development of a large variety of pharmacodynamic-monitoring strategies. The first reports of their clinical relevance are available, but further understanding of the analytical and clinical variables involved are required for the development of accurate, reproducible, and clinically relevant markers. PMID- 20009798 TI - Posterior fossa infarction following cleft palate repair and the arcuate foramen. PMID- 20009799 TI - Chest reconstruction: I. Anterior and anterolateral chest wall and wounds affecting respiratory function. AB - LEARNING OBJECTIVES: After studying this article, the participant should be able to: 1. Describe the indications for chest wall reconstruction. 2. Understand the function of the chest wall and implications for both reconstruction and the chest wall itself when components are missing or used for reconstruction. 3. List the reconstructive requirements of chest wall wounds. 4. Identify flaps for regional reconstruction of the chest wall. 5. Describe the role of microvascular surgery in chest wall reconstruction. BACKGROUND: Chest wall and mediastinum wounds may be life-threatening. They interfere with respiratory mechanics and may also be contaminated with exposed vital structures. Consideration is given to flap choice to restore function, resolve infection, and maintain suitable aesthetics. METHODS: Literature search as well as the authors' personal experience enabled preparation of this article. RESULTS: Where necessary, skeletal integrity must be restored, generally with prosthetic material, and then covered with well vascularized soft tissue. "Living tissue" is required to help combat infection, buttress visceral repairs, and fill dead space. Soft-tissue deficiency must occasionally be augmented with large distant microvascular flaps. CONCLUSION: Flap reconstruction has reduced morbidity and mortality of these complex problems without undue donor-site impairment of respiratory and upper extremity function. PMID- 20009800 TI - Nasal anatomy and John B. Deaver, M.D. PMID- 20009802 TI - Superficial parotidectomy for rhytidectomy contour refinement. PMID- 20009801 TI - A theory explaining the development of late-onset tightening or contracture in patients who have undergone facial reanimation surgery with free functional muscle transfers. PMID- 20009803 TI - A multicenter study of the efficacy and safety of restylane in the treatment of nasolabial folds in China. PMID- 20009804 TI - A new approach to the repair of partial mastectomy defects following conservation therapy of breast cancer located in the upper aspect of the breast. PMID- 20009805 TI - Lateral approach for internal mammary vessel dissection as a safer method in free flap breast reconstruction. PMID- 20009806 TI - Primary nipple reconstruction with AlloDerm: is a dermal flap always necessary? PMID- 20009807 TI - Breast implant in complex lymphatic malformation. PMID- 20009808 TI - Botulinum toxin type A infiltration for pain control after breast augmentation. PMID- 20009809 TI - Ulnar nerve penetration by a volar ganglion in the Guyon canal. PMID- 20009810 TI - Fibrosis of the hand secondary to jet propellant exposure: a new clinical syndrome. PMID- 20009811 TI - Long-term radiographic evaluation of AlloDerm reconstruction of the abdominal wall: material stretch as the cause of recurrence? PMID- 20009812 TI - External fixation in complicated groin flap surgery. PMID- 20009813 TI - Composite lower back and buttock reconstruction with gluteal myocutaneous flap in the just born. PMID- 20009814 TI - Abdominal wall imbrication using acellular dermal matrix: a novel technique for correcting laxity in post-bariatric surgery patients. PMID- 20009815 TI - A simple apparatus for pannus suspension during exceptionally large panniculectomy procedures. PMID- 20009816 TI - Antegrade venous drainage in a reverse-flow anterolateral thigh flap. PMID- 20009817 TI - Spindle cell melanoma arising from decades-old burn scar. PMID- 20009818 TI - Intralesional botulinum toxin type A injection as a new treatment measure for keloids. PMID- 20009819 TI - Evidence for a locally limited form of heparin-induced thrombocytopenia in plastic reconstructive surgery? PMID- 20009820 TI - Flipping and recruitment of instrumentally tied knots as a means of saving suture material. PMID- 20009821 TI - Breast cancer screening prior to cosmetic breast surgery: ASPS members' Adherence to American Cancer Society Guidelines. AB - BACKGROUND: The goal of this study was to determine the self-reported breast cancer screening practices of American plastic surgeons and the degree to which those practices adhere to the American Cancer Society guidelines. An independent analysis of subgroups divided by gender, years in practice, and practice setting was performed and the implications of the results are discussed. METHODS: The authors conducted an online survey of the members of the American Society of Plastic Surgeons. Questions assessed practice composition, American Cancer Society guideline familiarity, and preoperative breast cancer screening in patients seeking aesthetic breast surgery. Responses were summarized, subgroup comparisons were made, and logistic regression was used to determine predictors of physician practices. RESULTS: The 1066 respondents were predominantly male (82 percent) and consisted largely of private practitioners (73 percent). In total, 47 percent appeared to follow the American Cancer Society guidelines, while 64 percent claimed familiarity. Being male predicted more accurate guideline knowledge, but being female resulted in more aggressive screening and possibly more diagnoses. Number of years in practice and familiarity with the American Cancer Society guidelines also resulted in more perioperative diagnoses. CONCLUSIONS: Knowledge of the American Cancer Society guidelines is an essential component of effective cancer screening, but only two-thirds of plastic surgeons claim familiarity with them, and fewer than half report concordant practices. As plastic surgeons who often perform surgical procedures on the breast in women with no history of breast disease, we have an obligation to understand and apply consistent, reliable breast cancer screening practices to ensure the well-being of our patients. PMID- 20009822 TI - Carcinoma and atypical hyperplasia in reduction mammaplasty: increased sampling leads to increased detection. A prospective study. AB - BACKGROUND: Reduction mammaplasty for symptomatic macromastia or correction of asymmetry is performed more than 100,000 times per year in the United States. The reported incidence of occult breast cancer in reduction mammaplasty ranges from 0.06 to 4.6 percent. No standard pathology assessment for reduction mammaplasty exists. The authors evaluated the incidence of occult carcinoma and atypical hyperplasia in reduction mammaplasty specimens and identified clinical risk factors. Systematic sampling of additional tissue sections was instituted to evaluate the hypothesis that increased sampling would identify more significant pathologic findings. METHODS: All reduction mammaplasty specimens over a 20-month period at a single institution were prospectively examined. All specimens had baseline gross and microscopic evaluations, and then each was subjected to systematic additional sampling. The incidence of significant pathologic findings (carcinoma and atypical hyperplasia) was tabulated. Variables such as age and preoperative mammogram were examined. RESULTS: A total of 202 cases were evaluated. Significant pathologic findings (carcinoma and atypical hyperplasia) were present in 12.4 percent. The rate of carcinoma was 4 percent in all patients (6.2 percent in patients >or=40 years and 7.9 percent in patients >or=50 years). CONCLUSIONS: A significantly higher rate (12.4 percent) of significant pathologic findings was identified in this prospective study compared with published literature. None of the lesions was identified on preoperative mammogram. Age was significantly associated with significant pathologic findings. Increased sampling was associated with significant pathologic findings only in patients 40 years or older, indicating the need for thorough sampling of reduction mammaplasty specimens in patients older than 40. PMID- 20009823 TI - The impact of breast reduction surgery on low-back compressive forces and function in individuals with macromastia. AB - BACKGROUND: Macromastia can be a morbid condition causing affected women to endure shoulder pain, back pain, intertrigo, and shoulder grooving from the bra straps. Subjective symptoms are effectively relieved by reduction mammaplasty surgery. Reliable objective evidence is limited in evaluating benefits for this patient population. METHODS: Eleven women undergoing reduction mammaplasty were evaluated before surgery and 3 months after surgery. Evaluation modeled maximal compressive forces at the low back during an unconstrained lifting task using inverse dynamics and measured the subjective level of disability using the Functional Rating Index. RESULTS: The analysis of the kinematic and kinetic data demonstrated no significant difference in ankle, knee, and hip joint angles between pretests and posttests. In addition, the average maximum low-back compressive forces decreased 35 percent for the unconstrained lifting task, and the Functional Rating Index scores improved 76 percent following surgery. Greatest improvement was reported for the frequency of pain and travel. CONCLUSION: Reduction mammaplasty surgery produced objective improvement in low back compressive forces and patients' reported level of functional disability. PMID- 20009824 TI - Gracilis myocutaneous free flap in autologous breast reconstruction. AB - BACKGROUND: The gracilis myocutaneous free flap provides an alternative for autologous breast reconstruction. It avoids abdominal donor-site morbidity, allows for a quicker recovery, provides an alternative to the thin patient with a hidden and acceptable donor site, and allows for supine positioning for harvest and inset in a timely fashion. METHODS: A retrospective review was conducted of all autogenous postmastectomy reconstructions performed between January of 2005 and March of 2008. All patients receiving gracilis myocutaneous flap reconstruction for postmastectomy defects were included in this study. Office and hospital charts were reviewed. RESULTS: Twenty-seven gracilis flaps were performed during the study period. Average patient age was 50.4 years (range, 35 to 63.4 years), and average body mass index was 25.6 (range, 19.4 to 35.5). Of the 21 patients, 9.5 percent had hypertension, 19 percent smoked, none were diabetic, 14 percent were obese, and 4.8 percent had documented cardiovascular disease. Outcomes included a flap success rate of 100 percent, average operating time of 4.9 hours for unilateral (15 patients) and 6.7 hours for bilateral (six patients) flaps, intraoperative arterial thrombosis rate of 13.6 percent, average hospital stay of 3.75 days, major complication rate of 7.4 percent, and average follow-up of 7 months. CONCLUSIONS: The gracilis myocutaneous free flap provides an alternative breast reconstruction option for today's breast cancer patient. It allows for a quick harvest in the supine setting, creation of a moderate breast volume, consistent anatomy, and acceptable donor-site morbidity with good contour. PMID- 20009825 TI - Establishment of perforator flap programs for breast reconstruction: the New England program experience. AB - BACKGROUND: Patients seeking perforator flap breast reconstruction often face difficult choices if these procedures are not offered locally. To meet this demand, regional referral centers have been created to provide the latest procedures and to centralize cancer care. In 2004, the authors started the New England perforator flap program. The authors present a blueprint of their program and examine its impact on patient satisfaction and referral patterns. METHODS: A retrospective chart review was performed on all women who underwent postmastectomy reconstruction at Beth Israel Deaconess Medical Center from 1999 to 2008. A total of 977 patients were identified. Demographics, complications, referring patterns, and state of residence were examined. A validated questionnaire (76.4 percent response rate) was administered determining satisfaction, quality of life, and referral patterns. RESULTS: After program creation, there was an increase in the number of patients undergoing reconstruction from outside the state (10.6 percent versus 20.9 percent, p < 0.0001). There was also an increase in the number of outside referrals to the institution (53.8 percent versus 64.4 percent, p = 0.016). Perforator flap reconstruction had a higher referral rate from outside the referral base compared with nonperforator flap reconstruction (76.6 percent versus 52.0 percent, p < 0.0001). Finally, there was an increase in general patient satisfaction in perforator flap compared with nonperforator flap patients (82.7 percent versus 65.8 percent, p = 0.002). CONCLUSIONS: The creation of a perforator flap program can increase referrals to an institution and increase general patient satisfaction. With the new National Accreditation Program for Breast Centers, more institutions may consider developing these programs. The authors offer their blueprint as a potential model for program design. PMID- 20009826 TI - Sensibility following innervated free TRAM flap for breast reconstruction: Part II. Innervation improves patient-rated quality of life. AB - BACKGROUND: Restoring sensory innervation may be a useful adjunct in free flap head and neck reconstruction but, as yet, has not been shown to improve outcomes of breast reconstruction. The authors' previous study demonstrated objectively improved sensation in a group of innervated transverse rectus abdominis musculocutaneous (TRAM) flap breast reconstruction patients relative to noninnervated flaps. This study compared patient-rated outcomes of free TRAM breast reconstruction in innervated versus noninnervated flaps. METHODS: Twenty seven women were randomized prospectively to undergo either innervated or noninnervated free TRAM flap breast reconstruction. For innervated flaps, the T10 intercostal nerve was harvested with the TRAM flap and neurotized to the T4 sensory nerve at the recipient site. Three validated outcome tools were administered after surgery: the Medical Outcomes Study 36-Item Short Form Health Survey, the Body Image after Breast Cancer Questionnaire, and the Functional Assessment of Cancer Therapy-Breast. Results were correlated with previously reported objective sensibility outcomes. RESULTS: Eighteen of 27 women returned their questionnaires a mean 48 months after free TRAM flap reconstruction. Demographic analysis revealed no significant differences in patient age, height, smoking, radiation therapy, and nipple-areola complex reconstruction between randomized patient groups. There was a statistically significant improvement in all three measures in patients who were randomized to receive innervated free TRAM flaps compared with those receiving noninnervated flaps. CONCLUSION: This study demonstrates that innervation of free TRAM flaps used for breast reconstruction not only improves sensibility but also has a positive effect on patient-rated quality of life. PMID- 20009827 TI - Discussion. Sensibility following innervated free TRAM flap for breast reconstruction: Part II. Innervation improves patient-rated quality of life. PMID- 20009828 TI - Improvement of the survival of human autologous fat transplantation by using VEGF transfected adipose-derived stem cells. AB - BACKGROUND: The efficacy of autologous fat transplantation is reduced by fat absorption and fibrosis due to fat necrosis. Enhanced transplant neovascularization early after transplantation may reduce these outcomes. The authors asked whether cell and concomitant gene therapy using adipose-derived stem cells transduced with vascular endothelial growth factor (VEGF) improves fat transplant neovascularization and survival. METHODS: Human adipose-derived stem cells were expanded ex vivo for three passages, labeled with 1,1'-dioctadecyl 3,3,3',3'-tetramethylindocarbocyanine (DiI), and transduced with VEGF or left untransduced. Human fat tissues were then mixed with the DiI-labeled VEGF transduced adipose-derived stem cells, the DiI-labeled adipose-derived stem cells, the known vascularization-promoting agent insulin, or medium alone, and 18 nude mice were injected subcutaneously with all four preparations, with each of the four designated spots receiving one of these four mixtures in a random fashion. Six months later, transplanted tissue volume and histology were evaluated and neovascularization was quantified by counting the capillaries. RESULTS: Control transplant survival was 27.1 +/- 8.2 percent, but mixture with the VEGF-transduced and VEGF-untransduced stem cells significantly increased transplant survival (74.1 +/- 12.6 percent and 60.1 +/- 17.6 percent, respectively). Insulin was less effective (37.7 +/- 6.9 percent). Histological analysis revealed both types of transplants consisted predominantly of adipose tissue, unlike the control transplants, and had significantly less fat necrosis and fibrosis. The VEGF-transduced, adipose-derived stem cell-treated transplants had significantly higher capillary density than the other transplants and bore DiI-double-positive and CD31-double-positive cells (i.e., adipose-derived stem cell-derived endothelial cells). CONCLUSION: Adipose-derived stem cells together with VEGF transduction can enhance the survival and quality of transplanted fat tissues. PMID- 20009829 TI - Discussion. Improvement of the survival of human autologous fat transplantation by using VEGF-transfected adipose-derived stem cells. PMID- 20009830 TI - Application of botulinum toxin type A in myocutaneous flap expansion. AB - BACKGROUND: Although the use of the expanded myocutaneous flap has many advantages, the time course is prolonged. The net gain in surface area during acute expansion is insufficient. In this study, botulinum toxin type A was applied to shorten the flap expansion period while obtaining an adequate surface area that would meet surgical requirements. METHODS: Seven minipigs were used for the authors' experiments. Two sides of the dorsolumbar section in each pig were divided randomly into the botulinum toxin type A-treated and the saline-treated groups. Two 200-ml expanders were implanted in the submuscular pocket. Inflation began 2 weeks later, and the period of expansion was observed in a double-blind manner. Two weeks after the final inflation, the expansion area was measured, and a 10 x 6-cm myocutaneous flap was elevated and sutured in situ, and the contraction of the myocutaneous flap was observed. RESULTS: Botulinum toxin type A shortened the expansion time by 17 days (p < 0.001). The average increment of effective expansion area (p = 0.009) and the average recruitment area (p = 0.001) in the botulinum toxin type A-treated group were significantly higher than in the saline-treated group. Moreover, contraction in axial length and width of the botulinum toxin type A-treated group was lower than that of the saline-treated group at each time point following transposition (p < 0.001). CONCLUSIONS: Botulinum toxin type A could decrease the resistance to myocutaneous flap expansion, speed up the inflation, increase the expansion area, and reduce the contraction of the myocutaneous flap. It is a safe and convenient method with which to assist myocutaneous flap expansion. PMID- 20009831 TI - Short periodic applications of the vacuum-assisted closure device cause an extended tissue response in the diabetic mouse model. AB - BACKGROUND: The vacuum-assisted closure device is a widely used mechanical modulator of wound healing; however, the optimal time kinetics of application have not been determined. The objective of the study was to optimize the kinetics of vacuum-assisted closure application. METHODS: Full-thickness wounds in seven diabetic mice per study group were treated with either an occlusive dressing alone, the vacuum-assisted closure device for 6 or 12 hours, or the vacuum assisted closure device periodically for 4 hours every other day or continuously for 7 days. Wound closure and tissue response were evaluated by macroscopic, histologic, and immunohistochemical analyses on day 7. RESULTS: Wound closure was significantly faster after short initial vacuum-assisted closure (6-hour and 12 hour groups) when compared with continuous treatment. Increased granulation tissue formation was seen in the 12-hour group (2.4-fold increase) and in those treated periodically for 4 hours every other day (3.2-fold increase) compared with the dressing-alone controls. Significant stimulation of cell proliferation was seen after all vacuum-assisted closure patterns (3.6- to 5.3-fold increase), whereas angiogenesis was augmented only after the device was applied for either three times for 4 hours (4.3-fold) or continuously (4.7-fold) when compared with dressing-treated wounds. Treatment three times for 4 hours showed a superior angiogenic effect also when compared with short initial applications (6-hour and 12-hour groups). CONCLUSIONS: Short vacuum-assisted closure treatment induced an extended biological response in the wound. A total of 12 hours of periodically applied vacuum-assisted closure reached a similar wound tissue response as continuously applied vacuum-assisted closure for 7 days. These findings suggest new clinical approaches for mechanical wound-healing devices. PMID- 20009832 TI - TGF-beta1 RNA interference in mouse primary dura cell culture: downstream effects on TGF receptors, FGF-2, and FGF-R1 mRNA levels. AB - BACKGROUND: Transforming growth factor (TGF)-beta1 and fibroblast growth factor (FGF)-2 have both been shown to have significant roles in the regulation of murine calvarial suture fusion. Methods to decrease gene expression of these cytokines and their respective receptors have been established, but because of side effects, clinical applications are limited. In this study, the authors examined the effect of TGF-beta1-specific small interfering RNA (siRNA) on the messenger RNA (mRNA) expression of TGF-beta1, its TGF-betaR1 and TGF-betaR2 receptors, and FGF-2 and its R1 receptor in murine dura cells. METHODS: A primary dura cell line was established from CD-1 mice. Transfection efficiency using Lipofectamine was determined using BLOCKiT. Dura cells were transfected with serial concentrations of TGF-beta1 siRNA to determine the optimal dose. In subsequent experiments, cells were transfected with 16 nM TGF-beta1 siRNA and harvested on posttransfection days 4, 7, 10, and 14 for RNA isolation and quantitative polymerase chain reaction. RESULTS: Optimal inhibition of TGF-beta1 mRNA expression was achieved at 16 nM siRNA. On posttransfection day 4, TGF-beta1 mRNA levels were significantly decreased but returned to baseline by day 14. TGF betaR1 mRNA expression remained unaffected by transfection throughout the time course. However, TGF-betaR2, FGF-2, and FGF-R1 demonstrated significant inhibition of mRNA expression on posttransfection day 4. CONCLUSIONS: These results indicate that TGF-beta1 siRNA has the potential to alter the murine dura cytokines responsible for suture fusion in vitro. Manipulating underlying cranial suture biology with siRNA technology may ultimately allow control over suture fusion. This intervention may ultimately function as an effective adjunct to surgical intervention for craniosynostosis. PMID- 20009833 TI - Discussion. TGF-beta1 RNA interference in mouse primary dura cell culture: downstream effects on TGF receptors, FGF-2, and FGF-R1 mRNA levels. PMID- 20009834 TI - A new method for the second-stage auricular projection of the Nagata method: ultra-delicate split-thickness skin graft in continuity with full-thickness skin. AB - BACKGROUND: Staged auricular reconstruction remains mainstream among the various techniques of microtia reconstruction using autogenous costal cartilage. The initial stage involves fabrication and implantation of the cartilage framework, followed by projection of the reconstructed auricle in the second stage. During the projection stage, the line of incision is usually made close to the helical rim, from the superoanterior margin of the helical rim to the region of the lobule. Generally, a fascial flap is raised and covered over a cartilage block to project the auricle, and a skin graft is inset over the raw surface of the newly created postauricular sulcus. METHODS: The authors developed a new refinement for the second-stage auricular projection, whereby the skin cover for the raw surface over the posterior aspect of the auricle and the postauricular sulcus is an ultra delicate split-thickness skin graft raised in continuity with the full-thickness skin over the anterior aspect of the auricle. RESULTS: Incorporation of this new technique has minimized the visibility of suture lines and improved the appearance of the superior otobasion. In addition, the dimension of the skin cover required can be designed with greater precision. Postoperative outcomes using this new technique for auricular projection have been more than satisfactory. CONCLUSION: More favorable results that carry less surgical stigma can now be achieved in auricular reconstruction using this new modification of Nagata's two-stage method. PMID- 20009835 TI - Outcomes of direct muscle neurotization in pediatric patients with facial paralysis. AB - BACKGROUND: Multistage reconstructive procedures are often required to try to restore the emotional potential of human expression in cases of facial paralysis. In this study, the senior author's (J.K.T.) experience with the technique of direct nerve to muscle neurotization as a part of multistage facial reanimation procedures is presented. Age, denervation time, etiology of the lesion, previous reconstructive procedures, and types of muscles responsible for animation were analyzed to make evidence-based recommendations on the indications of the technique as well as its role and effectiveness in facial reanimation. METHODS: Retrospective review of 37 pediatric patients who underwent direct muscle neurotization took place. The patients were divided into three groups, depending on the region that direct neurotization was aiming to augment. Group A involved 28 patients for eye closure and blink, group B included 15 patients for smile, and group C included 19 patients for depressor augmentation. RESULTS: Twenty patients were female and 17 male. Patient age ranged from 1 to 16 years, with a mean age (+/-SD) of 9 +/- 2.8 years. Denervation time ranged from 3 months to 15.25 years, and the mean denervation time was 6.72 years. Electromyographic scoring of the neurotized muscles showed an overall mean improvement of 36 percent for eye closure, 34.25 percent for blink, 37 percent for smile augmentation, and 30 percent for depressor function restoration. CONCLUSION: Direct muscle neurotization has a valid role in pediatric facial reanimation procedures, as it augments and promotes expressivity. PMID- 20009836 TI - Effect of axonal load on the functional and aesthetic outcomes of the cross facial nerve graft procedure for facial reanimation. AB - BACKGROUND: To improve the ability to prognosticate the final surgery outcomes, this study was carried out to explore the correlation between the number of motor axons given to cross-facial nerve grafts for smile restoration and the aesthetic and functional outcomes. METHODS: Sixty-nine cases had adequate nerve biopsy specimens and were selected for the authors' study. Patient information was collected from chart review. Using Terzis' evaluation scale, smile functional and aesthetic outcomes as depicted in standardized videos were graded by a panel of four independent reviewers. Digital images of nerve specimens in stages I and II were obtained by using a microscope with a digital camera attachment. Using MetaMorph software, the number of motor axons was calculated, with the exception of the nerve specimens at the distal nerve grafts in stage II, which were quantitated manually. Mann-Whitney and Fisher's exact tests were used to test the effects of axon numbers and other factors on the outcomes. RESULTS: The donor axonal input correlated with the axon number at the distal end of the nerve graft and also correlated with the improvement of evaluation; however, no significance was found between the counts at the distal end of the nerve graft and the clinical outcomes. An important observation was that patients with a donor nerve count of 900 or higher showed a greater likelihood of achieving satisfactory results. CONCLUSION: The axon count at the donor nerve has a stronger influence on the final results. PMID- 20009837 TI - Visual assessment of linear scars: a new tool. AB - BACKGROUND: Existing scar evaluation tools are based on verbal descriptions and used primarily for burn scar assessment. To evaluate linear scars, the authors developed a new tool called the visual assessment of linear scars. This study was designed to determine whether patients and surgeons rated scars similarly and to test the intrarater and interrater reliability of the surgeons' ratings. METHODS: At 6 months postoperatively, 51 patients used the visual assessment of linear scars tool to rate each of their own surgical scars (two scars for abdominal; six scars for breast), and two surgeons used the visual assessment of linear scars tool to rate the patients' scar photographs. A post hoc two-sample t test was used to determine whether the raters' scar means were significantly different from each other. Spearman correlation was used to determine the intrarater and interrater reliability. RESULTS: There was no difference in the mean scar ratings between the surgeons; however, both surgeons had significantly higher mean ratings than the patients (a higher score is a worse-appearing scar). Intrarater reliability for the surgeons was large (rho > 0.5) and interrater reliability between the surgeons was also large (rho = 0.53) but borderline. CONCLUSIONS: The visual assessment of linear scars tool was developed to create a simple, straightforward method of assessing the overall appearance of the postsurgical linear scar, keeping in mind that the patient's perspective might differ from that of a surgeon or researcher. Both patients and surgeons found the ratings easy to perform, and the results showed that patients might rate their scars' appearance more favorably than the surgeons. The visual assessment of linear scars is a reliable tool with two plastic surgeons' rating of repeated photographs. PMID- 20009838 TI - Pectus excavatum and free fasciocutaneous infragluteal flap: a new technique for the correction of congenital asymptomatic chest wall deformities in adults. AB - BACKGROUND: Pectus excavatum is typically a cosmetic congenital chest wall deformity. In most cases, it does not affect heart and lung function; therefore, because of their high rate of complications, extensive procedures need not be performed. Various alternative techniques (e.g., reconstruction with silicone prosthesis or the transverse rectus abdominis musculocutaneous flap) were introduced in asymptomatic pectus excavatum. All of these methods have their advantages but also limitations. Thus, the authors used a free fasciocutaneous infragluteal flap for reconstruction of asymptomatic pectus excavatum in selected patients. METHODS: Between 2001 and 2007, six patients suffering from asymptomatic pectus excavatum underwent correction with the free fasciocutaneous infragluteal flap. The fasciocutaneous infragluteal flap is based on a constant end artery of the inferior gluteal artery. After raising of the flap and wound closure in the buttock region, the flap was adjusted to the defect using a small skin incision in the inframammary fold, and the vessels were anastomosed. RESULTS: There were no flap losses and no major complications. One patient suffered from a sensory change at the posterior thigh in the early postoperative period that resolved completely within 2 weeks. In four cases, flap shaping or liposuction was performed to improve the aesthetic result. In the authors' final evaluation, all patients were very satisfied with the result and would undergo the procedure again. CONCLUSIONS: The authors have demonstrated for the first time the reconstruction of asymptomatic pectus excavatum with the free fasciocutaneous infragluteal flap. It is the authors' opinion that, in selected patients, this flap offers an excellent alternative to established techniques for this problem. PMID- 20009839 TI - The perforasome theory: vascular anatomy and clinical implications. AB - BACKGROUND: A clear understanding of the vascular anatomy of an individual perforator relative to its vascular territory and flow characteristics is essential for both flap design and harvest. The authors investigated the three dimensional and four-dimensional arterial vascular territory of a single perforator, termed a "perforasome," in major clinically relevant areas of the body. METHODS: A vascular anatomy study was performed using 40 fresh cadavers. A total of 217 flaps and arterial perforasomes were studied. Dissection of all perforators was performed under loupe magnification. Perforator flaps on the anterior trunk, posterior trunk, and extremities were studied. Flaps underwent both static (three-dimensional) and dynamic (four-dimensional) computed tomographic angiography to better assess vascular anatomy, flow characteristics, and the contribution of both the subdermal plexus and fascia to flap perfusion. RESULTS: The perfusion and vascular territory of perforators is highly complex and variable. Each perforasome is linked with adjacent perforasomes by means of two main mechanisms that include both direct and indirect linking vessels. Vascular axis follows the axiality of linking vessels. Mass vascularity of a perforator found adjacent to an articulation is directed away from that same articulation, whereas perforators found at a midpoint between two articulations, or midpoint in the trunk, have a multidirectional flow distribution. CONCLUSIONS: Each perforator holds a unique vascular territory (perforasome). Perforator vascular supply is highly complex and follows some common guidelines. Direct and indirect linking vessels play a critical part in perforator flap perfusion, and every clinically significant perforator has the potential to become either a pedicle or free perforator flap. PMID- 20009840 TI - On the origins of components separation. AB - SUMMARY: The method of abdominal hernia repair known as components separation, described by Ramirez et al. in 1990, has enjoyed widespread popularity because of the sound anatomical principles it uses and most importantly the clinical success of the procedure. Although the authors who described this technique made no proprietary claims and did not discuss the history of abdominal hernia repair, they have certainly been credited with the development of this procedure. With this article, the author hopes to place components separation in historical perspective, and give credit to Donald Herron Young, who published the concept of external oblique relaxing incisions for repair of epigastric hernias in 1961. Born in Canada, Young studied and worked as a surgeon in England for most of his life. A biographical sketch is included. PMID- 20009841 TI - Barbed suture tenorrhaphy: an ex vivo biomechanical analysis. AB - BACKGROUND: Using barbed suture for flexor tenorrhaphy could permit knotless repair with tendon-barb adherence along the suture's entire length. The purpose of this study was to evaluate the tensile strength and repair-site profile of a technique of barbed suture tenorrhaphy. METHODS: Thirty-eight cadaveric flexor digitorum profundus tendons were randomized to polypropylene barbed suture repair in a knotless three-strand or six-strand configuration, or to unbarbed four strand cruciate repair. For each repair, the authors recorded the repair site cross-sectional area before and after tenorrhaphy. Tendons were distracted to failure, and data regarding load at failure and mode of failure were recorded. RESULTS: The mean cross-sectional area ratio of control repairs was 1.5 +/- 0.3, whereas that of three-strand and six-strand barbed repairs was 1.2 +/- 0.2 (p = 0.009) and 1.2 +/- 0.1 (p = 0.005), respectively. Mean load to failure of control repairs was 29 +/- 7 N, whereas that of three-strand and six-strand barbed repairs was 36 +/- 7 N (p = 0.32) and 88 +/- 4 N (p < 0.001), respectively. All cruciate repairs failed by knot rupture or suture pullout, whereas barbed repairs failed by suture breakage in 13 of 14 repairs (p < 0.001). CONCLUSIONS: In an ex vivo model of flexor tenorrhaphy, a three-strand barbed suture technique achieved tensile strength comparable to that of four-strand cruciate repairs and demonstrated significantly less repair-site bunching. A six-strand barbed suture technique demonstrated increased tensile strength compared with four-strand cruciate controls and significantly less repair-site bunching. Barbed suture repair may offer several advantages in flexor tenorrhaphy, and further in vivo testing is warranted. PMID- 20009842 TI - Refinements of tissue expansion for pediatric forehead reconstruction: a 13-year experience. AB - BACKGROUND: Reconstruction of the forehead in children when 25 percent or more of the forehead is involved presents a complex reconstructive challenge because of the confluence of highly visible aesthetic units. The present study was performed to develop an algorithm for lesions involving 25 percent or more of the forehead. METHODS: A 13-year retrospective review was performed of all pediatric patients who completed reconstruction for lesions involving at least 25 percent of the forehead by a single surgeon (A.K.G.). All lesions were classified on the basis of percentage of forehead involved and involvement of adjacent subunits. RESULTS: Twenty patients completed reconstruction. The median number of surgical procedures required was six (range, two to 11), with a median of three (range, one to four) expansion procedures. Simultaneous expanders were placed in the scalp (16 patients) and cheek (eight patients). Five patients underwent correction of eyebrow ptosis at a final procedure. Reconstruction involved 25 to 70 percent of the forehead in 19 patients, 17 of whom were reconstructed with serial forehead expansion and advancement flaps. One patient with a pigmented nevus occupying more than 75 percent of the forehead received an expanded full thickness skin graft from the lower abdomen. For all groups, the entire extent of the visible lesion was excised and complete skin coverage achieved. CONCLUSIONS: Reconstruction of 25 to 70 percent or more of the forehead in children is best accomplished using tissue expansion and direct advancement of adjacent tissues. Simultaneous expansion should be performed in the cheek and scalp if indicated. Brow ptosis should be addressed with each advancement. Lesions greater than 70 percent of the forehead are best accomplished with distant tissues. PMID- 20009843 TI - Reconstruction of extensive composite posterolateral mandibular defects using nonosseous free tissue transfer. AB - BACKGROUND: The management of composite oromandibular defects involving the posterolateral mandible and surrounding soft tissue remains a reconstructive challenge. Although bony reconstitution restores continuity of the mandible, osteocutaneous flaps sometimes do not provide adequate soft-tissue coverage of these postablative defects. The purpose of this study was to evaluate the use of soft-tissue flaps for extensive posterolateral oromandibular defects. METHODS: Consecutive patients who underwent reconstruction of composite oromandibular defects following posterolateral mandibulectomy between 1992 and 2006 were identified. Patient data were obtained from a prospectively maintained clinical database. Medical records were reviewed to characterize the extent of all postablative soft-tissue defects. Soft-tissue resection zones were defined as those involving the external cheek skin and/or lips, intraoral lining, tongue, retromolar trigone, palate, pharynx, and/or esophagus. RESULTS: In total, 76 patients were identified as having extensive posterolateral oromandibular defects reconstructed with soft-tissue flaps alone. In 62 percent of patients who underwent nonosseous free-tissue transfer, the oromandibular defect involved two or more soft-tissue zones. The most common flap used was the vertical rectus myocutaneous flap (n = 68). At the time of discharge, 54 percent of patients were on an oral diet. Sixty percent of patients had intelligible speech. Overall aesthetic outcome was good in 49 percent, fair in 21 percent, and poor in 30 percent of patients. CONCLUSIONS: Extensive composite defects of the posterolateral mandibular can be repaired effectively using soft-tissue flaps alone. When reconstructing a defect involving (1) the posterolateral mandible, overlying soft-tissues, and external skin and/or (2) the posterolateral mandible and two or more adjacent soft-tissue zones, the use of a soft-tissue flap alone can maximize success. PMID- 20009844 TI - The incidence of lower eyelid malposition after facial fracture repair: a retrospective study and meta-analysis comparing subtarsal, subciliary, and transconjunctival incisions. AB - BACKGROUND: Many incisional techniques have been described for access to the craniofacial skeleton for traumatic fracture repair, including subciliary, subtarsal, and transconjunctival approaches. Each is associated with a distinct set of complications. The goal of the authors' study was to examine lower lid malposition associated with these incisional approaches at the authors' institution and in the literature. METHODS: A retrospective chart review of all operative orbital and/or zygomatic fractures at the Beth Israel Deaconess Medical Center from 1998 to 2008 was performed. The surgical approach and postoperative lower eyelid complications were noted. A literature review was performed of all peer-reviewed studies in the English language examining the incidence of complications of lower eyelid incisions in operative facial trauma. RESULTS: A total of 180 cases were treated with a lower eyelid incision; these were identified as subtarsal (41 percent), subciliary (31 percent), and transconjunctival (25 percent). The risk of ectropion was highest in subciliary incisions (12.5 percent, p = 0.018); however, only two cases required operative management. Entropion was found in two cases after transconjunctival incisions; both required operative management. Seventeen studies were included in the meta analysis, representing 2086 patients. The risk of ectropion was highest in subciliary incisions (14 percent), the risk of entropion was highest in transconjunctival incisions (1.5 percent), and the risk of hypertrophic scarring was highest in subtarsal incisions (3.4 percent). A majority of these complications resolved with conservative management. CONCLUSIONS: Lower eyelid complications can be seen with all three incisional approaches to facial fracture repair. Based on their findings, the authors favor the subtarsal approach for zygomaticomaxillary fractures and the transconjunctival incision for isolated orbital floor fractures. PMID- 20009845 TI - Discussion. The incidence of lower eyelid malposition after facial fracture repair: a retrospective study and meta-analysis comparing subtarsal, subciliary, and transconjunctival incisions. PMID- 20009846 TI - Correction of secondary cleft lip deformity: the whistle flap procedure. AB - BACKGROUND: The Kapetansky flap was described to repair a whistle deformity or central vermilion defect following a primary bilateral cleft lip repair. The authors studied a modification of this technique, called the Whistle (wide-hinged island swing transposition labial enhancement) flap, to correct a wide array of secondary cleft lip deformities. METHODS: Patients with secondary cleft lip deformity who underwent the Whistle flap correction were studied (n = 21). Vertical lip height, lateral lip projections, and three-dimensional volumetric measurements were recorded along with physician and parent-patient satisfaction surveys. RESULTS: From 1994 to 2006, 23 Whistle flap procedures were performed on 21 cleft lip patients (bilateral, n = 13; unilateral, n = 8). The average age at surgery was 15.4 years (range, 13 to 21 years). The average follow-up was 27.6 months. The vertical height of the cleft side Cupid's bow to vermilion bottom had a mean increase of 164 percent. Lateral projection of the upper lip showed a mean increase of 117 percent. Preoperatively, patients exhibited mean volumetric asymmetry of 29.4 percent, and this was reduced following treatment to approximately 3.1 percent. Physician satisfaction using a modified Whitaker classification (categories I through IV) demonstrated that 56 percent of patients were category I (no refinements necessary) and 44 percent were category II (minor revisions are advisable). Parent-patient satisfaction in the follow-up period was a mean of 3.5 as measured using a five-point scale ranging from 0 to 4. There were minimal perioperative complications. CONCLUSION: The Whistle flap procedure provided a versatile and reliable option for the correction of vermilion defects from secondary cleft lip deformities. PMID- 20009847 TI - Differential diagnosis of the idiopathic laterally deviated mandible. AB - SUMMARY: Among patients who present for an orthognathic evaluation, a common finding is the presence of a laterally deviated mandible. At times, there is a cause to which the lower face asymmetry can be attributed, such as personal history of trauma to the face, presence of congenital disease known to affect the mandible, or other developmental abnormalities. More commonly, however, the patient is underdiagnosed and the cause of the idiopathic, laterally deviated mandible may be perplexing. The authors review the common causes of lateral deviation of the mandible and discuss how the growth of the mandible is affected and how the correct diagnosis may be ascertained. Among the common diagnoses, there are two major categories of mandibular asymmetry: (1) altered cranial base (e.g., muscular torticollis, unilateral craniosynostosis, and deformational plagiocephaly) and (2) condylar abnormality (e.g., condylar fractures, condylar hyperplasia, juvenile condylar arthritis, and hemifacial microsomia). Proper diagnosis and subsequent treatment of the underlying abnormality of the deviated mandible ensure the patient of the appropriate orthognathic reconstruction and decrease the likelihood of skeletal relapse after surgery. PMID- 20009848 TI - Primary repair of the bilateral cleft lip nose: a long-term follow-up. AB - BACKGROUND: The author previously published a method of primary repair of bilateral cleft lip nasal deformity using an open tip rhinoplasty to enable correction of the displaced and deformed alar cartilages. METHODS: The first 10 consecutive children who had this treatment have been followed through to adulthood. They are now 18 to 19 years old, and the long-term results of this primary nasal repair are reviewed. RESULTS: In general, the repair has been effective and stable. There has been no interference with growth. In most cases, the external nasal scars are barely noticeable. CONCLUSIONS: In primary treatment of the bilateral cleft lip nose, an open tip rhinoplasty enables accurate replacement of the alar cartilages and correction of the deformity. It justifies the creation of an external scar that is usually unnoticeable. PMID- 20009849 TI - Middle Eastern rhinoplasty in the United States: Part I. Primary rhinoplasty. AB - BACKGROUND: Middle Eastern rhinoplasty is becoming more frequent in the United States. Mature patients request a more refined, feminine nose, whereas younger patients (aged 15 to 25 years) want a smaller, "cuter" nose. METHODS: The author completed a prospective study of 50 consecutive female primary Middle Eastern rhinoplasty patients. The presenting anatomy, including the alar anatomy, and selected surgical techniques and their variations for particular cases were recorded. RESULTS: The majority of patients requested and received a significant change in their nasal appearance. Of the 40 patients followed for 18 months, three (7.5 percent) had revisions, two minor and one moderate. There were no functional complaints. CONCLUSIONS: The author emphasizes the need for structure throughout the nose, with use of both spreader grafts and columellar struts, to support the large and often heavy skin sleeve. Tip suture techniques, with or without tip grafts, are highly effective and there is no need for destructive tip techniques. PMID- 20009850 TI - Middle Eastern rhinoplasty in the United States: Part II. Secondary rhinoplasty. AB - BACKGROUND: There have been relatively few articles in the English language on secondary Middle Eastern rhinoplasty. This article analyzes the cause and treatment of secondary Middle Eastern rhinoplasty. METHODS: A prospective study of 40 consecutive female secondary Middle Eastern rhinoplasty patients was completed. RESULTS: The majority of secondary rhinoplasty patients were older than 25 years. Half of the patients had undergone a single prior rhinoplasty and the other half had undergone multiple operations, ranging in number from two to five. A wide variety of surgical techniques was necessary because of the broad range of presenting deformities, patients' requests, and the author's preferred procedures. CONCLUSIONS: The principal reasons for secondary rhinoplasty in Middle Eastern patients were a failure to correct the original deformity and the presence of visible surgical stigmata. The persistent complaints were a poorly defined tip and a long, droopy nose. Surprisingly, most secondary rhinoplasty patients had thin skin (55 percent), which necessitated fascia or dermis grafts to conceal surgical stigmata. At the time of secondary surgery, there was an absence of structure in these noses as evidenced by the prior 0 percent insertion of spreader grafts and the 10 percent use of columellar struts in prior open cases. Also, there was little evidence of other structural grafts, including alar rim, alar battens, or lateral crural strut grafts. Insertion of structural support appears essential to control primary deformities and to repair secondary deformities. PMID- 20009851 TI - Discussion. Middle Eastern rhinoplasty in the United States: Part II. Secondary rhinoplasty. PMID- 20009852 TI - An original application of the Endotine Ribbon device for brow lift. AB - BACKGROUND: Correction of brow ptosis and lateral temporal laxity is one of the goals of surgical rejuvenation of the aging face. The Endotine Forehead (Coapt Systems, Inc., Palo Alto, Calif.) device is an effective bioabsorbable fixation tool for forehead and brow-lift procedures. However, the Endotine Forehead device alone is not able to correct lateral brow ptosis and temporal laxity. In this article, the authors propose an innovative use of another special device, the Endotine Ribbon (Coapt Systems), to provide long-lasting results in lateral brow lift surgery and temporal lateral laxity correction. METHODS: Between February of 2006 and April of 2007, a total of 30 patients, aged between 38 and 70 years (average, 50 years), underwent brow-lift surgery. With this technique, the Endotine Ribbon is cut in halves and each portion of it is anchored to the deep temporal fascia with sutures, and its multiple tines facing outward grasp the superficial temporal fascia. The amount of brow elevation produced was assessed by comparison of the preoperative and postoperative vertical distances between the superior eyebrow hairline and the midpupil and lateral and medial canthal angle. The average follow-up period was 18 months. RESULTS: Using this technique, perfectly symmetric lateral eyebrows and temporal laxity correction were obtained in all patients. A lasting result was observed, and no significant adverse events were encountered. CONCLUSIONS: The application of the Endotine Ribbon for brow lift procedures provides significant and reproducible lateral brow elevation and temporal laxity correction. This fixation method is effective, safe, and easy to use, and leads to high patient satisfaction. PMID- 20009853 TI - Augmentation mammaplasty by reverse abdominoplasty (AMBRA). AB - BACKGROUND: The purpose of this article is to describe a novel technique of providing autologous tissues for breast augmentation and simultaneously rejuvenating the abdomen. METHODS: Thirty-seven patients underwent augmentation mammaplasty by reverse abdominoplasty (AMBRA) between 1997 and 2006. The upper abdominal pannus present in women whose lower abdomen was typically less aesthetically compromised was harvested as deepithelialized adipofascial flaps, maintaining their connection to and thus blood supply from the attached breast parenchyma. These flaps are transposed subglandularly, creating autologous tissue breast implants, and reverse abdominoplasty accomplishes donor-site closure and aesthetic improvement. If previous surgery or inadequate inframammary fold tissue thickness renders the superior circulation unfavorable, the upper abdominal tissues can be used as advancement flaps vascularly supplied by their attachment to the abdominal skin apron. RESULTS: Twenty-three patients (62 percent) had simultaneous mastopexy and 16 (43 percent) had simultaneous panniculectomy. Complications in the superior pedicle group were minimal. In the inferior pedicle group, complications were more extensive because of the premorbidity of this group of patients and the limitations of this technique, where the resuspension of the abdominal wall apron is less facile and generally weaker than closure with superiorly based flaps. CONCLUSIONS: Augmentation mammaplasty by reverse abdominoplasty is a versatile procedure that in the carefully selected patient can successfully address two aesthetic concerns simultaneously, providing durable autologous tissue that can obviate or enhance the outcome provided by prosthetic implants and rejuvenating the abdomen. It also shows promise as a significant adjunct to the techniques available to the breast reconstructive surgeon. PMID- 20009854 TI - Discussion. Augmentation mammaplasty by reverse abdominoplasty (AMBRA). PMID- 20009855 TI - A comparative analysis and systematic review of the wound-healing milieu: implications for body contouring after massive weight loss. AB - BACKGROUND: Wound-healing complications following body contouring for massive weight loss patients are significant, with rates exceeding 40 percent. To better understand aberrant healing in this population, the authors have performed a comparative analysis of the wound milieu literature for patient populations with similar complication rates. METHODS: PubMed and Ovid databases were reviewed from January of 1985 to January of 2009 for key terms, including wound healing, obesity, cancer, burn, transplant, and body contouring. Serum and wound levels of multiple factors, including matrix metalloproteinases (MMPs) and cytokines, were assessed. RESULTS: Complication rates in body contouring surgery range from 31 to 66 percent. Sixty-five studies were reviewed, and wound-healing complication rates were identified for cancer (45.8 percent), burn (30.4 percent), posttransplant (36 percent), and obese (43 percent) populations. In these groups, matrix metalloproteinases and tissue inhibitors of metalloproteinase (TIMPs) help regulate wound repair. Matrix metalloproteinase levels were elevated in cancer (4 fold increase in MMP-2), burn (20- to 30-fold increase in MMP-9), transplant (1.4 fold increase in MMP-2), and obese/chronic (79-fold increase) populations. TIMPs were increased in cancer (1.9-fold increase in TIMP-2) and burn (1.4-fold increase in TIMP-1) patients but decreased in chronic wound (55-fold decrease in TIMP-1) populations. Alterations to these regulatory proteins lead to prolonged matrix degradation, up-regulation of inflammatory mediators, and decreased growth factors, delaying the wound-healing process. CONCLUSIONS: Complications after body contouring surgery are likely multifactorial; however, molecular imbalances to the massive weight loss wound milieu may contribute to poor surgical outcomes. Examining wound regulatory proteins including transforming growth factor-beta, vascular endothelial growth factor, and matrix metalloproteinases could aid in understanding the healing difficulties observed clinically. PMID- 20009856 TI - Discussion. A comparative analysis and systematic review of the wound-healing milieu: implications for body contouring after massive weight loss. PMID- 20009857 TI - Liposuction abdominoplasty: an advanced body contouring technique. AB - BACKGROUND: Liposuction abdominoplasty was first performed by the author in 1997. In 2002, the procedure was presented as an "evolving concept" at the 71st Annual Meeting of the American Society of Plastic Surgeons. Over the next 6 years, an additional 294 procedures were added to the initial 43 (n = 337 patients), culminating in an advanced body contouring technique. METHODS: The advanced technique is characterized by a hydro-lipo-dissection technique that is selectively performed on the deep abdominal fascia and the Scarpa fascia and along the excision lines. Selective release of the skin-retaining ligaments preserves perforators and elevates a large island flap, extending from the pubis to the inframammary fold and from one flank to the other. Perforating neurovascular bundles tether the flap, minimizing dead space and providing improved flap sensibility. Diastasis recti repair is performed using newly devised segmental techniques. The flap possesses a robust circulation that enables upright patient ambulation and concomitant liposuction of circumferential torso and additional areas (322 patients). Surgery is ambulatory and performed under local anesthesia with monitored intravenous sedation, drains are not used, and patients are discharged 2 to 3 hours after surgery. RESULTS: There were no major complications, deaths, or deep vein thrombosis. Five late infections of seromas (1.4 percent) responded to local drainage, and six marginal necroses (1.7 percent) healed without ill effect. CONCLUSION: Over the past 11 years, liposuction abdominoplasty has established new concepts and evolved into an advanced body contouring technique. PMID- 20009858 TI - Preoperative use of alpha tocopherol does not increase the risk of hematoma in the face lift patient: a preliminary report. AB - BACKGROUND: Antiplatelet therapy and nutritional supplements are factors that may increase the risk of hematoma after routine face lift. This has led to the often repeated instruction "do not take any vitamins, herbs, aspirin, and any medications that may increase bleeding for 10 days before your face lift operation." At odds and diametrically opposed to this goal is combating the increased risk of deep venous thrombosis and venous thromboembolism associated with surgery. METHODS: On the basis of clinical observations, the author has incorporated the routine use of preoperative oral alpha tocopherol (vitamin E) into his preoperative instructions. The cohort included 146 patients scheduled for elective face lift enrolled in this study from February of 2007 to April of 2008. Aspirin and clopidogrel were stopped before surgery. Patients were instructed to take 400 IU of alpha tocopherol daily 3 weeks before surgery. RESULTS: No hematoma developed in any of the 146 patients. The time for routine drainage was not prolonged. No clinically evident case of deep vein thrombosis or venous thromboembolism occurred. The clinical impression was that increased bruising was not observed. CONCLUSIONS: A recent landmark article has documented the efficacy of alpha tocopherol in the prevention of venous thromboembolism in over 39,000 patients. The addition of this readily available, over-the-counter medication may have profound effects in the prevention of a potentially devastating complication in the elective cosmetic surgery patient. Future prospective studies may further elucidate the efficacy of alpha tocopherol in the prevention of venous thromboembolism. PMID- 20009859 TI - Toward outcomes-based plastic surgery training: a needs assessment of recent graduates. AB - BACKGROUND: The Accreditation Council for Graduate Medical Education Outcomes Project mandates data-driven improvements to training programs. Few outcomes measures exist in plastic surgery. The purpose of this study was to determine how well we presently train plastic surgery residents within the context of the council's competencies and how well those competencies are practiced after residency. METHODS: Plastic surgery graduates from 2000 to 2005 were surveyed using a new questionnaire organized around the accreditation council's competencies. Scores from individual item responses were averaged to create a composite score reflecting the quality of training and practice of each competency. Composite scores were then compared using nonparametric analyses. Data regarding currently used assessment tools were also measured. RESULTS: The response rate was 21 percent. The respondent sample was representative of current plastic surgeons. Overall, the competencies were considered important, with increasing awareness in more recent years. Practice-based learning and improvement appeared to be the best taught, and systems-based practice appeared to be the most poorly taught. There were differences between independent and integrated formats but they did not support one over the other. Women did not rate their training as highly as men did in some areas, and open comments suggested there were significant issues at some programs. CONCLUSIONS: Recent graduates view plastic surgery training as excellent. The competency domain in greatest need of attention is systems-based practice. We must broaden our use of assessment instruments for more valid results and investigate gender differences in our training programs. These results describe the outcomes of current plastic surgery training and serve as a needs assessment for improvement and as a baseline for future comparison. PMID- 20009860 TI - A systematic review of ethical principles in the plastic surgery literature. AB - BACKGROUND: This purpose of this study was to perform a systematic review to identify articles that discuss ethical issues relating to the field of plastic and reconstructive surgery and to evaluate whether ethical issues are underrepresented in the plastic surgery literature. METHODS: Four medical databases were selected to search through the medical literature with specific inclusion criteria to disqualify irrelevant articles from the study. Appropriate articles were extracted, and their quality and validity were assessed by multiple investigators to maximize reproducibility. The data were then synthesized and analyzed for associations among the ethical principles. RESULTS: Of a total library search of more than 100,000 plastic surgery-oriented articles, only 110 clearly focused on ethical principles. Autonomy (53 percent) was the most common major theme, whereas distributive justice (15 percent) represented the least frequently emphasized ethical principle. The proportions of each ethical principle were tested against each other for equality using Cochran's Q test; the Q test reached statistical significance (Q = 67.04, df = 3, p < 0.0001), indicating that the ethical principles were not discussed equally in plastic surgery literature, which was expected because autonomy represented 53 percent of the articles, whereas distributive justice represented only 15 percent of articles. When examining both major and minor themes, more than half of the articles (61 percent) addressed two or more ethical principles. Beneficence and nonmaleficence were strongly associated (Pearson's chi = 55.38, df = 1,p < 0.0001). CONCLUSION: Despite the extensive number of ethical issues that plastic surgeons face, a relatively small proportion of plastic surgery literature was dedicated to discussing ethical principles. PMID- 20009861 TI - So you want to be an expert. PMID- 20009867 TI - An examination of posttraumatic, postsurgical orbital deformities: conclusions drawn for improvement of primary treatment. PMID- 20009868 TI - Anatomical variations of the occipital nerves: implications for the treatment of chronic headaches. PMID- 20009869 TI - Breast cancer: do local failures affect survival? PMID- 20009871 TI - Techniques of monitoring buried fasciocutaneous free flaps. PMID- 20009872 TI - Revitalizing the training of clinical scientists in surgery: a U.K. perspective. PMID- 20009873 TI - Aesthetic surgery's tipping point. PMID- 20009876 TI - Current world literature. PMID- 20009880 TI - CYP1A1 and CYP2D6 gene polymorphisms in Israeli Jewish women with cervical cancer. AB - HYPOTHESIS: To assess the frequency distribution of the CYP1A1 and CYP2D6 gene polymorphisms in Israeli Jewish women with cervical cancer. METHODS: Forty-three Israeli Jewish women with cervical cancer and 123 healthy Israeli Jewish women were assessed. CYP1A1 (2 alleles) and CYP2D6 polymorphism was analyzed using an allele-specific, polymerase chain reaction-based method. RESULTS: The allele frequency and genotype distribution for the CYP1A1 (2 alleles) and CYP2D6 polymorphism did not reach a level of significance. Smoking was the only independent risk factor for cervical cancer (P = 0.0003). CONCLUSION: CYP1A1 and CYP2D6 mutations are not related to an increased risk for cervical cancer in the Jewish Israeli population. PMID- 20009881 TI - Gain of the human telomerase RNA gene TERC at 3q26 is strongly associated with cervical intraepithelial neoplasia and carcinoma. AB - This study investigated the gain of the human telomerase RNA gene TERC at 3q26 in patients with uterine cervix disease in the southern part of China and assessed the relationship between TERC gain and cervical pathological findings. One hundred ten cervical specimens, which were collected from patients with various kinds of uterine cervix disease that was subsequently diagnosed as chronic cervicitis and with examination results negative for intraepithelial lesion or malignancy (NILM, n = 23), mild dysplasia (cervical intraepithelial neoplasia type 1 [CIN1], n = 37), moderate dysplasia (CIN2, n = 12), severe dysplasia (CIN3, n = 10), and squamous cell carcinoma (SCA, n = 28) confirmed by histologic diagnosis, were analyzed for the proportion of abnormal cells with TERC gain using a commercially available 2-color fluorescence in situ hybridization (FISH) probe. The cases with a higher proportion of abnormal cells than the threshold evaluated by NILM were recorded as positive TERC gain. The chi and Kruskal-Wallis tests were used to assess the associations between FISH findings and diagnoses. The incidence of positive TERC gain in the cases diagnosed as NILM or CIN1 was significantly lower than that in cases diagnosed as CIN2, CIN3, or SCA (P < 0.01). In addition, a significantly higher proportion of abnormal cells with TERC gain was found as a pathological change from CIN1 to SCA (P < 0.01). We conclude that the TERC gain seems to be an important associated genetic event in CIN and carcinoma; FISH is a potential tool for the diagnoses of uterine cervix disease. PMID- 20009882 TI - Lymph node involvement in epithelial ovarian cancer: sites and risk factors in a series of 355 patients. AB - OBJECTIVES: To perform a cartography of lymph node metastases in epithelial ovarian cancer and to determine predictive factors of lymph node metastases. METHOD: The charts of 355 patients with epithelial ovarian cancer who underwent lymphadenectomy during a primary (n = 252) or secondary debulking surgery (n = 103) were analyzed. The topography of the lymph node metastases was notified for the whole group according to the stage of the disease, the histological type, and the moment of surgery. In patients who underwent a primary surgery before chemotherapy, independent prognostic variables for the risk of lymph node involvement were tested with a multivariate analysis. Independent prognostic factors were combined to determine risk profiles in individual patients. RESULTS: The main area of the lymph node metastases was para-aortic. Isolated pelvic lymph node involvement was 10%. Three variables independently predicted lymph node invasion: advanced T stage, high-risk histological profile, and metastases. CONCLUSIONS: When lymphadenectomy is recommended, systematic lymph node dissections in the aortic and pelvic areas are warranted. An isolated pelvic lymph node assessment, particularly in the early stages, is inappropriate. By combining independent risk factors, a useful tool for individual risk assessment of lymph node involvement could be established, helping to decide whether to perform a lymph node dissection, especially at restaging surgery. PMID- 20009883 TI - Risk of hospitalizations associated with adverse effects of chemotherapy in a large community-based cohort of elderly women with ovarian cancer. AB - PURPOSE: To determine the rates of hospitalization after receiving chemotherapy in patients who were diagnosed with ovarian cancer and to examine significant predictors for hospitalizations. METHODS: We studied 9361 women who were diagnosed with stages I to IV ovarian cancer at age 65 or older in 1991 to 2002, identified from the 16 areas of the Surveillance, Epidemiology, and End Results program linked with Medicare data. Hospitalization for adverse effects associated with chemotherapy was defined using primary and secondary diagnosis codes from the inpatient claims. Multivariate logistic regression was used to estimate the risk of being hospitalized for adverse effects in patients receiving chemotherapy compared with those who did not. RESULTS: A total of 1363 patients (14.6% of 9361) received platinum-based chemotherapy without taxane, 3094 patients (33.1%) received platinum-taxane combination chemotherapy, 1694 patients (18.1%) administered other (nonplatinum) chemotherapy, and 3210 patients (34.3%) did not receive chemotherapy. Compared with those receiving platinum-based chemotherapy, patients receiving nonplatinum chemotherapy had a higher risk of being hospitalized for infection (odds ratio [OR], 1.66; 95% confidence interval [95% CI], 1.19-2.31), whereas patients who did not receive chemotherapy (OR, 0.16; 95% CI, 0.10-0.28) or received platinum-taxane combination chemotherapy (OR, 0.54; 95% CI, 0.34-0.86) were significantly less likely to be hospitalized for hematologic toxicities. Although both comorbidity scores and age were significant predictors for hospitalization for infection and cardiovascular diseases, older age was not a significant predictor for gastrointestinal and hematologic toxicities. CONCLUSIONS: The nonplatinum chemotherapeutic regimens were associated with higher rates of hospitalizations for gastrointestinal and hematologic conditions or infections compared with platinum-based or platinum taxane combination regimens. Comorbidity was a significant predictor for hospitalization for infections and gastrointestinal and cardiovascular diseases. PMID- 20009884 TI - Expression of p53 protein phosphorylated at serine 20 and serine 392 in malignant and benign ovarian neoplasms: correlation with clinicopathological parameters of tumors. AB - INTRODUCTION: The modification of p53 protein by phosphorylation plays an important role in its stabilization and the regulation of its biological properties. The study investigated the expression of p53 protein phosphorylated at serine 20 (Ser20) and Ser392 and the association between clinicopathological parameters of ovarian neoplasms with respect to p53 protein overexpression. METHODS: p53 protein expression was evaluated on tissues from malignant and benign ovarian tumors. Protein expression was measured in a subset of the specimens using immunohistochemistry. RESULTS: The correlation between p53 protein overexpression and p53-Ser392 phosphorylation was found in ovarian carcinomas (P = 0.001, r = +0.27). In the total group of ovarian carcinomas, significant differences were observed in p53 protein overexpression between well (G1) and poor (G3) tumor grades (P = 0.005) and between serous and endometrioid types of tumor (P = 0.04), whereas p53-Ser20 phosphorylation was associated with advanced International Federation of Gynecology and Obstetrics stage (P = 0.004) and high tumor grade (P = 0.02). In p53-positive ovarian carcinomas, p53-Ser392 phosphorylation was associated with advanced tumor stage (P = 0.02) and high tumor grade (P = 0.049). p53-Ser20 phosphorylation was associated with low tumor grade of p53-positive ovarian carcinomas (P = 0.02) and with high tumor grade of p53-negative ovarian carcinomas (P = 0.02). CONCLUSIONS: These results revealed that p53 phosphorylation at Ser20 and Ser392 is an early event in ovarian tumor development. The authors suggest that the expression of p53 protein phosphorylated at Ser20 and Ser392 in ovarian carcinomas determines their individual clinical features depending on p53 protein status and may be useful biological biomarkers characterizing their behavior. PMID- 20009885 TI - Study of T lymphocytes infiltrating peritoneal metastases in advanced ovarian cancer: associations with vascular endothelial growth factor levels and prognosis in patients receiving platinum-based chemotherapy. AB - INTRODUCTION: The presence of CD3(+) tumor-infiltrating lymphocytes (TILs) has been found to correlate with improved survival in epithelial ovarian cancer, but the association of TIL subpopulations with clinical outcome remains controversial. We performed a prospective analysis of TIL subpopulations from patients with epithelial ovarian cancer and their activation status and studied their association with prognosis. METHODS: Flow cytometric analysis was performed on TIL subpopulations isolated from 45 fresh ovarian tumor specimens, obtained during surgery, after mechanical dissociation and enzymatic degradation. Vascular endothelial growth factor and tumor necrosis factor alpha levels in ascites and serum were measured by enzyme-linked immunosorbent assay. RESULTS: Significantly increased numbers of CD56(+) cells (natural killer and natural killer-like T cells; P = 0.045), activated CD4(+)HLA-DR cells (P = 0.046), and activated CD8(+)CD25(+) cells (P = 0.028) were found in serous and endometrioid carcinomas compared with mucinous and clear cell carcinomas. A high percentage of CD4(+)CD25(hi) cells (regulatory T cells) and activated CD4(+)HLA-DR cells significantly associated with improved median overall survival (not reached vs 35 months [P = 0.0241] and not reached vs 35 months [P = 0.0144], respectively) and median progression-free survival (30 months vs 14 months [P = 0.0819] and 30 months vs 13 months [P = 0.0479], respectively). Vascular endothelial growth factor ascites levels were inversely correlated with CD14(+) (rho = -0.529, P = 0.001), whereas HLA-DR8(+)CD8 lymphocytes were inversely correlated with both ascites and serum vascular endothelial growth factor levels (rho = -0.494, P = 0.006, and rho = -0.586, P = 0.037, respectively). CONCLUSIONS: The presence of regulatory T cells and activated CD4(+) cells within the tumor microenvironment is associated with improved overall and progression-free survival in patients with ovarian cancer. PMID- 20009886 TI - Preoperative CA-125 levels in 123 patients with borderline ovarian tumors: a retrospective analysis and review of the literature. AB - We studied preoperative CA-125 levels of 123 patients with borderline ovarian tumors (BOTs) and performed an analysis with data of earlier published studies. CA-125 levels were compared according to histology and stage of disease. Preoperative serum CA-125 levels were significantly higher for patients with advanced stage (median, 181 U/mL; range, 413 U/mL) compared with patients with stage I (median, 28 U/mL; range, 1123 U/mL) BOTs and for patients with serous (median, 59 U/mL; range, 1119 U/mL) compared with patients with mucinous (median, 25 U/mL; range, 371 U/mL) BOTs (both P < 0.001, Mann-Whitney U test). A pooled analysis of 3 studies and the present study showed positive rates of CA-125 (value >35 U/mL) in 171 (53%) of 325 patients with BOTs. Positive rates were more often found in patients with serous (67%) compared with patients with mucinous BOTs (39%) and in patients with advanced stage (83%) compared with patients with stage I BOTs (47%) (both P < 0.001, Pearson chi(2) test). This main effect was also found for each individual study of the pooled analysis. From a clinical perspective, we believe, on base of the results of this study and the literature, that preoperative discrimination using CA-125 level is especially difficult between patients with stage I ovarian cancer and the group of patients with serous and/or advanced-stage BOTs. PMID- 20009887 TI - Overexpression of clusterin in ovarian cancer is correlated with impaired survival. AB - Clusterin has been found to be overexpressed in several human malignancies and also be expressed in ovarian carcinoma tissues. However, to date, no study has investigated the prognostic significance of clusterin expression in ovarian carcinoma. Therefore, we examined the relationship between clusterin overexpression and clinicopathological features to determine its prognostic relevance. Eighty-six patients diagnosed with primary ovarian cancer between 1993 and 2004 were selected and recorded follow-up data and clinicopathological data. The expression of clusterin was detected on the sections of tissue microarray by immunohistochemistry and was evaluated the association with patient's clinical features and prognosis. Overexpression of clusterin protein in ovarian cancer was observed in 46.5% of the patients and was found more often in disease that was in the advanced stage (P = 0.0001). The expression levels of clusterin was associated with International Federation of Gynecology and Obstetrics stage (P = 0.0001) and histologic type (P = 0.002). However, no significant association was observed between clusterin expression and patient age or tumor Silverberg grade (P > 0.05). In addition, the average survival time of the patients with clusterin overexpression was significantly shorter than that with normal expression of clusterin. Clusterin expression was associated with survival of patients with primary ovarian cancer (relative risk for overall survival 1.69; 95% confidence interval, 1.52 to 1.95 (P = 0.033)). Our data show that clusterin is not only a biomarker associated with ovarian cancer, but it also appears to be a prognostic factor associated with adverse outcome. PMID- 20009889 TI - The impact of complete surgical staging upon survival in early-stage ovarian clear cell carcinoma: a multi-institutional retrospective study. AB - Pure-type clear cell carcinoma (CCC) has been recognized as a distinct subtype of ovarian cancer, showing resistance to conventional platinum-based chemotherapy and resulting in poor prognosis. The aim of the study was to evaluate the effects of complete surgical staging procedures for early-stage CCC patients in a retrospective multi-institutional analysis. During the period 1992 to 2002, a total of 199 patients with pT1 M0 CCC were identified. Survival analysis was estimated by Kaplan-Meier methods, and prognostic factors were evaluated using a Cox regression model. Among pT1 M0 tumors, retroperitoneal lymph node status was negative in 125 cases (pN0, 63%), positive in 10 cases (pN1, 5%), and unknown in 64 cases (pNx, 32%). Progression-free survival of pN1 was significantly worse than that of pN0 (P < 0.05), whereas there was no significant difference between pN1 and pNx. There was no significant difference of overall survival (OS) among the 3 groups. Multivariate analysis revealed that peritoneal cytology status was the only independent prognostic factor for progression-free survival (P = 0.04), but completion of surgical staging procedures was not a prognostic factor. There was no significant prognostic factor for OS. Our study implied that complete surgical staging enabled us to distinguish a high-risk group of recurrence in pT1 M0 CCC; however, the procedure could not improve OS. Although the study was a limited retrospective study, the impact of peritoneal cytology status was more important than complete surgical staging procedure in CCC patients. More effective treatment modality was warranted, especially for CCC cases positive for malignant peritoneal cytology. PMID- 20009888 TI - HOXA10 is overexpressed in human ovarian clear cell adenocarcinoma and correlates with poor survival. AB - Human homeobox gene (HOX) A10 is a homeobox allotype gene of the HOXA family in the HOX family. Human homeobox gene A10 may play an important role in cancer development. However, the role of HOXA10 in the carcinogenesis of ovarian clear cell adenocarcinoma (OCCA) has not been established. We have evaluated the prognostic significance of HOXA10 expression for human OCCA and the effects of HOXA10 on proliferation, motility, and invasion of OCCA cells. We found that HOXA10 was not expressed in normal ovarian epithelium, ovarian endometrial cysts, and ovarian serous carcinomas, but 20 (68.9%) of the 29 OCCAs were positive for the expression of HOXA10. Human homeobox gene A10 expression was negatively correlated to the 5-year survival of OCCA patients (R = -0.442, P = 0.043). When a HOXA10 expression vector was stably transfected into a human OCCA cell line, ES 2, the proliferation rate of ES-2-HOXA10 was much higher than the vector control, the motility of ES-2-HOXA10 cells was significantly increased compared with the control (P < 0.05), and the invasion of ES-2-HOXA10 cells was also much higher than the vector control (P < 0.01 at 5 hours and 12 hours after scratching). In conclusion, HOXA10 was overexpressed in OCCA and was correlated with poor survival. HOXA10 promotes proliferation, migration, and invasion of OCCA cells. Human homeobox gene A10 could be a promising prognostic marker for OCCA. PMID- 20009890 TI - Distribution of second primary malignancies suggests a bidirectional effect between breast and endometrial cancer: a population-based study. AB - INTRODUCTION: The aim of this study was to investigate the incidence of second primary tumors in patients with breast cancer (BC), with particular regard to bidirectional risk for endometrial cancer (EC). METHODS: A total of 7512 and 343 patients with first and second primary BC, respectively, were referenced to the expected number of cases calculated using the standardized incidence ratio (SIR) over the same period, to evaluate the observed and expected ratio between the groups. Data on tamoxifen use were also considered. RESULTS: A total of 499 women with primary BC developed a second tumor. The total SIR, that is, the ratio between observed second primary cancer among patients with BC and the expected primary cancers in the general population, was significantly higher (SIR = 1.23; 95% confidence interval, 1.12-1.34; P = 0.007), particularly for melanoma (2.25), EC (2.15), ovarian cancer (1.74), hematologic malignancies (1.36), and bilateral BC (1.25). A greater risk of BC after thyroid (2.22) and EC (1.62) was also observed. Furthermore, the risk of developing EC was higher in patients treated with tamoxifen (SIR = 2.50 vs 1.34). CONCLUSIONS: Bidirectional risk of endometrial cancer was not exclusively related to tamoxifen use. PMID- 20009891 TI - Pure papillary serous tumors of the endometrium: a clinicopathological analysis of 61 cases from a single institution. AB - OBJECTIVE: The aim of this study was to evaluate the clinicopathological data and prognosis factors corresponding to patients with papillary serous carcinoma of the endometrium treated at a single institution. METHODS: Medical and anatomopathological records were reviewed in the Department of Gynecological Oncology of the Canarian University Hospital between 1989 and 2006. Only pure cases of papillary serous carcinoma of the endometrium were included. Survival rates were analyzed using the Kaplan-Meier technique. RESULTS: The study included 61 patients. Stages I, II, III, and IV were identified in 32.8%, 19.7%, 31.1%, and 8.2% of patients, respectively. Event-free survival rates after 5 and 10 years were 59% and 40%, respectively, with a median time of 83 months (95% confidence interval, 63-110). Relapse occurred in 40.5% of the patients. Overall survival rates after 5 and 10 years were 37.7% and 29%, respectively, with a median time of 55 months (95% confidence interval, 40-70). A univariate analysis showed that prognosis factors were tumor markers, stage, myometrial infiltration, lymphovascular invasion, and ganglia involvement. A multivariate analysis showed that tumor markers, stage, and myometrial infiltration had an independent influence on overall survival. CONCLUSIONS: Papillary serous carcinoma of the endometrium is a very aggressive type of endometrial carcinoma that behaves more similar to ovarian carcinoma than to endometrial carcinoma. Tumor markers at diagnosis, stage, and myometrial infiltration mainly determine prognosis at our institution. PMID- 20009892 TI - Laparoscopic versus abdominal hysterectomy for endometrial cancer: comparison of patient outcomes. AB - OBJECTIVE: To compare the demographics, cancer characteristics, and hospital outcomes of endometrial cancer patients undergoing a laparoscopically assisted vaginal hysterectomy (LAVH) versus a total abdominal hysterectomy (TAH). METHODS: Two California population databases (Office of Statewide Health Planning and Development and the California Cancer Registry) were linked using patient identifiers. Patients who underwent endometrial cancer surgery from 1997 to 2001 were identified. The combined database was queried for type of surgery, patient demographics, hospital outcomes, comorbidities, and cancer characteristics. Statistical analyses included the t test, chi2 test, and logistic regression. RESULTS: In this study, 978 endometrial cancer patients (7.7%) had an LAVH and 11,765 (92.3%) had a TAH. The mean ages for the 2 groups were 63.3 and 64.8 years, respectively. Lymphadenectomy was performed more frequently in LAVH patients compared with TAH patients (45.6 vs 41.1%; P = 0.006). Patients undergoing LAVH were more likely to be younger and healthier and have stage I or grade 1 disease (P < 0.0001). Total abdominal hysterectomy patients were more likely to have significant medical comorbidities. Mean length of stay for LAVH was 2.40 versus 4.36 days for TAH (P < 0.001), but mean hospital charges were comparable. Perioperative complications such as vascular and bowel injuries, pulmonary embolism, wound problems, and transfusions were significantly more common in TAH patients. CONCLUSION: Surgeons seem to carefully select endometrial cancer patients for laparoscopic surgery. Although surgical staging was performed in less than 50% of endometrial cancer patients, the rate was not worse in laparoscopic procedures. Short-term hospital complications were less common in the laparoscopy group. PMID- 20009893 TI - Kallikrein 4 overexpression in endometrial carcinoma and upregulation by estrogen via mitogen-activated protein kinase signal pathway. AB - OBJECTIVE: The aim of this study was to investigate the expression of kallikrein 4 (KLK4) and the potential signal pathway through which estrogen up-regulates KLK4 in endometrial cancer. METHODS: The expression of KLK4 was analyzed in 15 human normal endometrium, 13 hyperplasia endometrium, and 68 endometrioid adenocarcinoma by immunohistochemistry. After exposure to 17beta-estradiol and/or to the mitogen-activated protein kinase (MAPK) inhibitor U0126 and to the PI3K inhibitor LY294002, the expression of KLK4 in the endometrial cancer cell lines KLE and RL95-2 was detected with quantitative reverse transcriptase polymerase chain reaction (qRT-PCR) and Western blot. RESULTS: The expression of KLK4 protein was higher in endometroid endometrial cancer than in hyperplasia or normal endometrium (P < 0.001). Immunohistochemical staining revealed that 92.6% (63/68) of endometrial adenocarcinoma, 61.5% (8/13) of hyperplasia endometrium, and 26.7% (4/15) of normal endometrium were positive for KLK4 protein. The expression of KLK4 was significantly associated with tumor grade (P = 0.004), but not with ER status (P = 0.532). Quantitative reverse transcriptase PCR and Western blot analysis showed that estrogen can up-regulate the expression of KLK4 in endometrial cancer cell lines KLE and RL95-2, and the up-regulation effect of 17beta-estradiol on KLK4 can be inhibited by U0126 in the 2 endometrial cancer cell lines but not by LY294002. CONCLUSIONS: Kallikrein 4 is a new nuclear protein, and estrogen up-regulates the expression of KLK4 by activating the MAPK pathway in endometrial cancer cell lines, which may play an important role in the development of endometrial cancer. PMID- 20009894 TI - Prognostic significance of preoperative thrombocytosis in patients with endometrial carcinoma in an inner-city population. AB - INTRODUCTION: Thrombocytosis is present in a wide range of malignancies, with a reported incidence of 10% to 57%. Several reports have documented thrombocytosis at the time of diagnosis as a poor prognostic indicator. Our study is the first report evaluating the role of preoperative thrombocytosis and its association with survival in a predominantly African American and Caribbean American urban population. MATERIALS AND METHODS: We retrospectively reviewed the charts of 99 consecutive patients treated for endometrial carcinoma at SUNY Downstate Medical Center. Seventy-seven patients were deemed eligible for the study, and the following clinicopathologic characteristics were recorded from their medical records: age, stage, grade, histological subtype, presence of lymphovascular space invasion, depth of myometrial invasion, intrauterine tumor volume, preoperative prothrombin time, activated partial thromboplastin time, platelet count, progression-free survival (PFS), and overall survival (OS). The data were analyzed using Spearman and Pearson correlations, Student t test, chi(2) test, and Fisher exact test. Survival analysis was performed using Kaplan-Meier tables, log-rank test, and Cox proportional hazard model. The 2-tailed value of P < 0.05 was considered significant. RESULTS: Fourteen (18.2%) of 77 patients exhibited thrombocytosis (platelet count, >400 x 10(9)/L). Patients with advanced disease (stages III-IV) had a significantly higher mean preoperative platelet count (359 +/- 23.8 x 10(9)/L) in comparison with patients with localized disease (stages I II, 283 +/- 14.3 x 10(9)/L, P = 0.005). The median PFS among patients with stages III and IV without preoperative thrombocytosis was 15.0 +/- 4.8 months (n = 21) and with thrombocytosis was 3.0 +/- 1.4 months (n = 8, P = 0.032). The median OS in patients without thrombocytosis was 24.0 +/- 4.5 months (n = 21) and in patients with thrombocytosis was 7.0 +/- 3.8 months (n = 8, P = 0.015). Multivariate analysis was performed using log-rank test and Cox proportional hazard model. The only variables that retained independent prognostic significance were stage (hazards ratio, 3.268; P = 0.040) and preoperative thrombocytosis (hazards ratio, 1.714 per 100 platelets; P = 0.030). Among patients with localized disease, preoperative thrombocytosis was not associated with worsened OS or PFS. CONCLUSIONS: Our data indicate that preoperative thrombocytosis among high-risk inner-city patients with stages III to IV endometrial cancer is an independent prognostic indicator. This is the first such report in a predominantly African American and Caribbean American population. Further research is needed to elucidate the mechanisms of thrombocytosis in malignancy. Association of thrombocytosis and aggressive tumor behavior warrants investigation of antiplatelet therapy and its effect on outcome. PMID- 20009895 TI - Early cervical cancer managed by laparoscopy and conventional surgery: comparison of treatment results. AB - INTRODUCTION: The goal of this retrospective analysis was to compare the results of treatment in patients with early cervical cancer managed by laparoscopy with those for patients observed after laparotomy. METHODS: The retrospective analysis was carried out with 22 patients operated on with total laparoscopic hysterectomy and 58 patients treated by abdominal hysterectomy. Patients with clinical tumor stage IA, IB1, or IIA were eligible for surgery. The main efficacy end point was disease-free survival evaluated by the Kaplan-Meier method. The survival curves were compared using log-rank tests. In addition, length of hospitalization, duration of surgery, and complication rate were compared. P < 0.05 was set as statistically significant. RESULTS: Predicted 3-year disease-free survival rates in the "open surgery" and "laparoscopy" groups were 0.86 (standard deviation [SD], 0.049) and 0.82 (SD, 0.098), respectively (P = 0.53). Recurrence rate was 13.6% after laparoscopy and 12% in open surgery. In 2 patients, intraperitoneal spread occurred after laparoscopy. The operation time was longer and hospitalization shorter after laparoscopy. CONCLUSIONS: The 3-year disease-free survival was similar in both groups. Total laparoscopic radical hysterectomy may be an option in early cervical cancer; however, the intraperitoneal spread in 2 patients compels a search for possible risk factors in patients managed by laparoscopy. PMID- 20009896 TI - Clinical significance of human papillomavirus genotype by linear array assay in Japanese women with uterine cervical lesions and type 16 physical status by in situ hybridization. AB - The aim of this study was to clarify the relation of human papillomavirus (HPV) genotypes and physical status in the cervical neoplasm of Japanese patients with the grade of the disease. Human papillomavirus genotype was detected using a linear array genotyping assay. Human papillomavirus status, diffuse or punctate signal pattern, was studied by biotynyl-tyramide-based in situ hybridization for positive cases of HPV-16. Human papillomavirus types 16, 52, 58, and 31, in descending order of frequency, were prevalent. The rates of HPV infection in patients with cervical intraepithelial neoplasia (CIN) or squamous cell carcinoma (SCC) were significantly higher than those in patients without cervical lesions. The frequency of HPV single infection in SCC was higher than that in CIN1 or CIN2. In an unspecified-risk HPV, types 66 and 70 were found in SCC and 62, 71, and 82 were detected in CIN3. The diffuse pattern was more frequent in CIN, and the punctate pattern was more frequent in SCC. Human papillomavirus types 16, 52, 58, and 31 were frequently detected in Japanese women with cervical neoplasias, and several unspecified-risk HPVs might be high-risk types. A single infection of HPV and a punctate signal pattern seemed to be closely correlated with cervical carcinogenesis. PMID- 20009897 TI - Increasing brachytherapy dose predicts survival for interstitial and tandem-based radiation for stage IIIB cervical cancer. AB - PURPOSE: To evaluate factors affecting survival and toxicity in patients with stage IIIB cervical cancer treated with external-beam radiotherapy and low-dose rate brachytherapy. MATERIALS AND METHODS: Seventy patients with stage IIIB cervical cancer treated between 1980 and 2000 were identified. A total of 51 patients had a tandem placed; 19 had interstitial brachytherapy only with no tandem placed. Univariate and multivariate analyses assessed factors potentially associated with survival and long-term complications. RESULTS: Kaplan-Meier estimates for overall survival (OS) for the entire population at 2, 3, and 4 years were 43%, 29%, and 27%, respectively. Overall survival differed significantly between those treated with a tandem versus those treated without (log rank P < 0.0001). Significant predictors of OS on univariate Cox regression were presence of a tandem (hazards ratio [HR], 0.31; P = 0.0001) and cumulative dose to point A (HR, 0.95; P = 0.0001). Multivariate regression showed that the presence of a tandem was the only significant predictor of OS (HR, 0.46; P = 0.04). There were no significant predictors of complications. CONCLUSIONS: The dose to point A of approximately 85 Gy as associated with tandem use was a significant predictor of survival in patients with stage IIIB cervical carcinoma, indicating the significance of the central location of the high dose provided by the tandem. Patients selected for interstitial brachytherapy alone must receive sufficient dose, and a tandem should be placed whenever feasible for all patients with an intact uterus treated for locally advanced cervical cancer. PMID- 20009898 TI - Abdominal radical trachelectomy in fertility-sparing treatment of early-stage cervical cancer. AB - BACKGROUND: Abdominal radical trachelectomy (ART) is one of the fertility-sparing procedures in women with early-stage cervical cancer. In comparison with vaginal radical trachelectomy, the published results of ART are so far limited. METHODS: Enrolled were women referred for ART either by laparoscopy or laparotomy. The main inclusion criterion was stage IA2 or IB1 with a cranial extent that allows for preservation of at least 1 cm of the endocervical canal. RESULTS: A total of 24 women were referred for the procedure, but fertility could not be preserved in 7 (29%) of them. Four women underwent immediate completion of radical hysterectomy because of a positive cranial surgical margin (n = 2) or sentinel node macrometastasis (n = 2) on frozen section. We found no correlation between tumor volume and inability to preserve fertility. A positive sentinel node was identified in 4 patients (17%); there were no false-negative results. Of the 9 women (53%) who have tried to conceive so far, 6 (67%) have conceived and 5 given birth, 2 of which were premature deliveries. CONCLUSIONS: Fertility cannot be preserved because of positive cranial margins or involved lymph nodes in almost one third of patients originally referred for radical trachelectomy. The main criterion for the selection of suitable patients should be the cranial extent of the tumor. Abdominal radical trachelectomy allows for achievement of satisfactory obstetrical outcomes. PMID- 20009899 TI - 18F-fluorodeoxyglucose-positron emission tomography and pathologic tumor size in early-stage invasive cervical cancer. AB - PURPOSE: Cervical cancer tumor size determined by 18F-fluorodeoxyglucose-positron emission tomography (FDG-PET) thresholding of the maximum standardized uptake value (SUVMax) has been correlated with the determined tumor size by computed tomography and magnetic resonance imaging. The purpose of this study was to evaluate the relationship between preoperative FDG-PET tumor size and pathologic tumor diameter in patients with early-stage cervical cancer who were undergoing radical hysterectomy. PATIENTS AND METHODS: Forty patients with early-stage cervical cancer underwent pretreatment FDG-PET/computed tomography before radical hysterectomy and lymph node dissection. Primary tumor diameter was defined on FDG PET as the largest diameter of the 3-dimensional volumetric isocontour of the 40% threshold of the SUVMax. The FDG-PET measurements were compared with the tumor diameter and the histological diameter of the pathologic specimen using regression analysis, paired t test, and unpaired t test. RESULTS: The FDG-PET tumor diameter measurements were correlated to the pathologic tumor diameter in the surgical specimen with a coefficient of determination (R2) of 0.951 and a correlation coefficient of 0.757 (P < 0.0001). CONCLUSION: There is a high level of correlation in the FDG-PET and the pathologic tumor measurements in the early stage cervical cancer. PMID- 20009900 TI - Human papillomavirus types 16 and 18 mRNA levels and not DNA levels may be associated with advancing stages of cervical cancer. AB - OBJECTIVE: Human papillomavirus (HPV) contributes to the development of cervical cancer. We hypothesize that HPV DNA and messenger RNA (mRNA) levels may be associated with increasing stages of cervical cancer. MATERIALS AND METHODS: In this study, we measured DNA and mRNA viral loads of the most common high-risk HPV 16 and HPV-18 in cervical biopsy tissue of women with cervical neoplasia using real-time polymerase chain reaction. RESULTS: Median HPV-16 and HPV-18 DNA viral loads were 58,342 copies and 71,367 per 5000 cells, respectively. We found that HPV-16 and HPV-18 DNA levels did not correlate with advancing tumor stage (P = 0.977 and P = 0.263). Messenger RNA transcripts were detected in 81 (86%) of HPV 16 DNA-positive women and in 16 (84.2%) of HPV-18-positive women. Median HPV-16 and HPV-18 transcript copy numbers were 5964 and 6158, respectively. In women with squamous cell carcinoma, HPV-16 mRNA loads showed an increasing but not statistically significant trend with advancing disease stage (rho = 0.231, P = 0.058). CONCLUSIONS: We conclude that HPV mRNA levels and not DNA levels may be associated with advancing stages of cervical cancer. PMID- 20009901 TI - Association of constitutive nuclear factor-kappaB activation with aggressive aspects and poor prognosis in cervical cancer. AB - INTRODUCTION: Although nuclear factor-kappaB (NF-kappaB) is generally believed to be involved in carcinogenesis, the relationship between NF-kappaB activation and progression of cervical cancer in clinical settings has not been reported. In this study, we investigated the association of NF-kappaB activation with aggressive aspects and prognosis in cervical cancer. METHODS: Nuclear factor kappaB subunits p65 and p50 were detected in 159 paraffin tissues including normal cervical, precancerous (squamous intraepithelial lesions), and cervical carcinoma tissues by immunohistochemistry. Nuclear factor-kappaB nuclear translocation and DNA-binding activity in precancerous or carcinoma tissues were examined by Western blot and electrophoretic mobility shift assay, respectively. RESULTS: A gradual NF-kappaB activation from normal cervical epithelial cells to precancerous and carcinoma cells was detected by immunohistochemistry (nuclear expression of p65 and p50, P < 0.001), Western blot (NF-kappaB nuclear translocation), and electrophoretic mobility shift assay (enhanced DNA-binding activity). In 79 cancer tissues, increased nuclear p65, an active NF-kappaB form, was correlated with poor tumor grade, lymphatic metastasis, interstitial invasion, and larger tumor size (P < 0.05). Similarly, increased nuclear p50 was correlated with poor tumor grade, interstitial invasion, and larger tumor size (P < 0.05). Moreover, increased nuclear p65 was associated with lower survival rate in patients with cancer (P < 0.05). CONCLUSIONS: Constitutive NF-kappaB activation is correlated to tumor progression, aggressive behaviors, and poor prognosis in cervical cancer, suggesting that NF-kappaB is a tumor promoter, a prognostic indicator, and a possible therapeutic target for this malignant disease. PMID- 20009902 TI - Epidemiological characteristics related to treatment failure of preinvasive cervical intraepithelial neoplasia among Brazilian women. AB - OBJECTIVE: To evaluate cervical intraepithelial neoplasia (CIN) treatment failure among women with an altered Papanicolaou test and treated at a public cervical pathology center in Rio de Janeiro, Brazil. METHODS: An exploratory study was carried out in a cohort of women treated for preinvasive cervical cancer, from 1998 to 2000. Epidemiological, clinical, and sociodemographic data were obtained from medical records. Information related to cytological and histological examinations, classified according to the Bethesda System, were cross-tabulated with reports from the main cervical pathology laboratory at the Brazilian Cancer Institute. Conditional probability for treatment failure (12 and 24 months) and hazard ratios were estimated through Kaplan-Meier (log-rank test, 95%) and Cox regression (entry, or=0.10). RESULTS: Mean (SD) follow-up period was 20.75 (20.53) months, and 449 women were treated for CIN-1 or CIN-2/3. There was an increased probability of treatment failure according to age group (37.19% and 57.44% in 12 and 24 months for women >50 years). Women who smoked for more than 10 years were more likely to present treatment failure compared with those who smoked for up to 10 years (33.56% and 22.01% in 12 months; 53.15% and 42.59% in 24 months, respectively). Women older than 50 years showed an independent risk for treatment failure (adjusted hazard ratio, 1.59; 95% confidence interval, 1.04-2.41) compared with women aged between 10 and 29 years. CONCLUSIONS: Age (>50 years) was statistically associated with treatment failure. Duration of tobacco use (>10 years), CIN-2/3 cytology at treatment, glandular involvement, and having more than 4 sexual partners in a lifetime were also associated with failure, reducing statistical significance after adjustment. PMID- 20009903 TI - Preventing cervical cancer: stakeholder attitudes toward CareHPV-focused screening programs in Roi-et Province, Thailand. AB - OBJECTIVE: To assess which of the 5 CareHPV-inclusive protocols stakeholders in Roi-et Province, Thailand found (1) most preferable and (2) most beneficial to the overall goal of reducing cervical cancer. DESIGN: Five CareHPV-inclusive cancer prevention approaches were presented to a convenience sample of colposcopists, trainers, health care providers, district medical directors, and district health officers. Participants ranked their preference for each plan (A E) compared with the current screening protocol and also the perceived comparative benefit of the plans. Plans differed in whether every patient was screened using both the human papillomavirus (HPV) test and visual inspection with acetic acid (VIA) or only HPV-positive women were screened with VIA; in clinician versus self-swab and in hospital/clinic-based or village-based screening. RESULTS: Overall, participants supported an innovative plan in which women would be screened in their homes and villages using the self-swab version of the CareHPV DNA test, and only those who screened positive for HPV are screened with VIA. When results were stratified by provider type (physician vs nonphysician) and/or practice concentration (hospital-oriented vs field oriented), preference and perceived benefit for the plans differed significantly with physician/hospital-oriented participants and are more likely to be skeptical of the self-swab version of the CareHPV test. CONCLUSIONS: Providers in Thailand recognize the potential value of a rapid HPV test as part of a cervical cancer prevention program and, overall, support a patient-administered self-swab followed by visual confirmation and treatment as the most practical CareHPV inclusive approach largely because of the likely increase in coverage. Future studies will further define the clinical benefits and limitations of this test. PMID- 20009904 TI - Can we diagnose invasive cervical cancer during pregnancy as precise as in nonpregnant women?: maternal and perinatal outcome in pregnancies complicated with cervical cancers. AB - Cervical cancer is the most common gynecologic malignancy associated with pregnancy. However, there are no consensus guidelines that define the indications for or the optimal length of expectant management. The subjects were women who had a preexisting invasive cervical cancer or whose cancers were diagnosed during pregnancy or within 12 months after delivery. Thirty-nine consecutive women with cervical cancer, whose ages ranged from 20 to 40 years, were chosen as controls. We performed a retrospective chart review on the maternal profile and perinatal outcome and compared the clinical features between pregnancy- and non-pregnancy associated cervical cancer in patients. The percentage of asymptomatic cases in which cancer was detected in a routine Papanicolaou test was significantly higher in the pregnant patients. The percentage of induced preterm labor or therapeutic abortions was 50%. Expectant management (mean length, 19.8 weeks) was chosen by 5 patients, and there were no cases of recurrence or death from disease. Seven subjects, including 5 patients whose diagnoses were changed from cervical intraepithelial neoplasm or condyloma to cancer, were managed as "unexpected expectant" because these subjects were not diagnosed as having stage IA/IB cancer during pregnancy. All of these subjects underwent vaginal delivery and included 2 patients with death from disease and lymph node recurrence. The percentage in which disease severity was underestimated was higher in pregnant patients. The option of therapeutic delay should be carefully discussed. Patient counseling should address the issue that risk may not be precisely estimated because of the possibility that disease severity may be underestimated during pregnancy. PMID- 20009905 TI - Phase 2 trial of docetaxel, gemcitabine, and oxaliplatin combination chemotherapy in platinum- and paclitaxel-pretreated epithelial ovarian cancer. AB - BACKGROUND: This phase 2 trial was designed to evaluate the efficacy and toxicity of a combination of docetaxel, gemcitabine, and oxaliplatin for platinum- and paclitaxel-pretreated epithelial ovarian cancer. PATIENTS AND METHODS: Heavily pretreated patients (N = 30; median age, 61 years) received docetaxel, 55 mg/m2; gemcitabine, 500 mg/m2 (day 1); and oxaliplatin, 70 mg/m2 (day 2) biweekly. Twelve patients had platinum-sensitive disease, and 18 patients had platinum resistant disease. RESULTS: Median follow-up was 18.6 months. No differences in patient characteristics were observed between patients with carboplatinum sensitive and carboplatinum-resistant disease. In patients with carboplatin sensitive disease, an overall response (OR) of 83.3%, a progression-free survival of 10.6 months, and an overall survival of 18.9 months were observed. In patients with carboplatinum-resistant disease, an OR was seen in 38.9% with a progression free survival of 5.3 months and an overall survival of 16.3 months. Patients with platinum-refractory disease (progression under previous carboplatinum therapy, n = 13) had an OR of 23%, whereas patients with objective response but relapse less than 6 months after carboplatinum therapy had an OR of 80.0%. Grade 3 and 4 toxicities were only observed for anemia (6.7%), neutropenia (20.0%), thrombopenia, peripheral neuropathy, and diarrhea (16.7%). No neutropenic fever or treatment-related death occurred. CONCLUSIONS: In comparison with current standard protocols, a combination of docetaxel, gemcitabine, and oxaliplatin showed considerably higher efficacy without remarkable increased toxicity; particularly for patients with early relapse after a platinum-containing therapy. PMID- 20009906 TI - Mastectomy approach with Y-shaped incision: a technique designed for women with obesity. AB - OBJECTIVE: To evaluate the access to axilla, postoperative complications, and cosmetic results of the modified radical mastectomy with a Y-shaped approach especially designed for women with obesity. METHODS: One hundred seventeen consecutive women with obesity with infiltrating breast cancer were studied. All of them were not eligible for breast-conserving therapy and underwent modified radical mastectomy. Operation was performed using a surgical technique designed to improve the axillary clearance and to eliminate the lateral dog ear deformity. Two oblique incisions were added to the traditional transverse Stewart incision at the lateral part forming the Y-shaped approach. After lateral flap retraction, the axillary dissection was done. Before closing the wound, the triangular flap was advanced medially, whereas superior and inferior areas of redundant skin overlying the latissimus dorsi muscle were excised. RESULTS: No intraoperative complications were observed. In each case, the axillary dissection (with level 3 node clearance when needed) was performed with ease. The wound was healed by primary adhesion, giving an excellent cosmetic result without lateral dog ear deformity. Skin flap necrosis was found in 2 elderly patients. Wound hematoma and surgical site infection developed in 1 patient each. Necrosis of the apex of axillary triangle occurred in one woman with diabetes. These rare complications were managed successfully in all the cases. CONCLUSIONS: The Y-shaped approach for modified radical mastectomy is a simple and safe technique. It facilitates the wide access to axilla and improves cosmesis in women with obesity by eliminating lateral dog ear deformity. PMID- 20009907 TI - Learning curve of laparoscopic radical hysterectomy with pelvic and/or para aortic lymphadenectomy in the early and locally advanced cervical cancer: comparison of the first 50 and second 50 cases. AB - BACKGROUND: To compare the surgical and oncological outcomes and morbidity of the first 50 cases treated by laparoscopic radical hysterectomy with those of the second 50 cases. METHODS: Between October 1994 and January 2004, we retrospectively reviewed the charts of 100 consecutive patients (International Federation of Gynecology and Obstetrics stages IA2 [n = 12], IB1 [n = 56], IB2 [n = 15], IIA [n = 15], and IIB [n = 2]) who underwent laparoscopic radical hysterectomy with pelvic and/or para-aortic lymphadenectomy. One hundred patients were divided into the first 50 cases (group 1) and second 50 cases (group 2). RESULTS: Operating time, length of hospital stay, time to normal residual urine, and transfusion rate significantly decreased, and the acquired number of pelvic nodes significantly increased when comparing group 1 with group 2. The intraoperative and postoperative complication rates profoundly decreased in group 2 as compared with group 1. After a median follow-up of 66.5 months, 10 patients had a recurrence, 9 of whom died. The 5-year overall survival rates were 96% in group 1 and 90% in group 2, and 5-year disease-free survival rates were 92% in group 1 and 90% in group 2. CONCLUSIONS: Laparoscopic radical hysterectomy is a feasible and safe treatment modality in early and even locally advanced cervical cancer without decreasing survival. Surgical outcome was improved with experience, and the complication rate related to operation of group 1 was higher than that of group 2. There was no significant difference in survival between the 2 groups. PMID- 20009908 TI - Biostatistics: a toolkit for exploration, validation, and interpretation of clinical data. AB - Biostatistics plays a key role in all phases of clinical research starting from the design to the monitoring, data collection, data analysis, and interpretation of the results. A clear understanding of the statistical framework as it relates to the study hypothesis, reported results, and interpretation is vital for the scientific integrity of the study and its acceptance in the general medical community. In this brief report, we will put in perspective the general analytical framework for exploring and validating prognostic factors using data from large databases. PMID- 20009909 TI - ALK gene rearrangements: a new therapeutic target in a molecularly defined subset of non-small cell lung cancer. AB - Transforming rearrangements of the ALK (anaplastic lymphoma kinase) gene have recently been described in non-small cell lung cancer (NSCLC). The most common rearrangement arises from an inversion in the short arm of chromosome 2 that creates a fusion between the 5' portion of the EML4 (echinoderm microtubule associated protein-like 4) gene and the 3' portion of the ALK gene. At least seven ALK gene rearrangement variants have been described involving different EML4-ALK breakpoints or rarely other non-EML4 fusion partners. ALK rearrangements may be readily identified in tumor tissue by reverse transcription-polymerase chain reaction or fluorescent in situ hybridization. Although ALK gene rearrangements affect only about 4% of all lung cancers, they are more frequent in adenocarcinomas, in never or light smokers, and seem almost mutually exclusive with activating EGFR or KRAS mutations. Promising results seen in patients with NSCLC containing fluorescent in situ hybridization-detected ALK rearrangements treated on a phase I study with PF02341066, an oral ALK inhibitor, indicate that ALK represents a new therapeutic target in this molecularly defined subset of NSCLC. PMID- 20009911 TI - The role of histology with common first-line regimens for advanced non-small cell lung cancer: a brief report of the retrospective analysis of a three-arm randomized trial. AB - INTRODUCTION: Although histology has not consistently been associated with treatment outcome in advanced non-small cell lung cancer, a recent phase III trial comparing pemetrexed plus cisplatin and gemcitabine plus cisplatin (GC) demonstrated better efficacy for pemetrexed plus cisplatin in nonsquamous (adenocarcinoma and large cell carcinoma) carcinoma than in squamous cell carcinoma. Herein, retrospective analysis is used to explore the potential predictive and prognostic role of non-small cell lung cancer histology in patients treated with three first-line, platinum-based regimens. METHODS: Survival and time to progression (TTP) data from a phase III trial comparing paclitaxel plus carboplatin (PCb), GC, and vinorelbine plus cisplatin (VC) were analyzed. Using Cox multiple regression, factors for one model included treatment (PCb, GC, and VC), histology (squamous, adenocarcinoma, large cell, and other), gender, Eastern Cooperative Oncology Group performance status (0/1 and 2), stage (IIIB and IV), number of metastatic sites (< or = 1 and >1), and smoking history (yes or no). In another model, histology was simply considered as squamous versus nonsquamous. An interaction value of p < 0.10 was considered significant. RESULTS: Baseline patient and disease characteristics for the 607 treated patients were balanced among the arms. No significant treatment-by-histology interaction was seen in either model for either end point. Nevertheless, histology was a significant prognostic factor for survival in the first model (p = 0.0183) and marginally significant for TTP (p = 0.0783). Subsequent pairwise comparisons of histology groups demonstrated a survival advantage for squamous cell carcinoma over adenocarcinoma (p = 0.0021). CONCLUSIONS: Histology was not predictive of PCb, GC, or VC treatment effect for either survival or TTP. Histology was prognostic for survival, with better outcomes associated with squamous cell carcinoma. PMID- 20009910 TI - Induction of E-cadherin in lung cancer and interaction with growth suppression by histone deacetylase inhibition. AB - INTRODUCTION: Loss of E-cadherin confers a poor prognosis in lung cancer patients and is associated with in vitro resistance to endothelial growth factor receptor inhibitors. Zinc finger E box-binding homeobox (ZEB)-1, the predominant transcriptional suppressor of E-cadherin in lung tumor lines, recruits histone deacetylases (HDACs) as co-repressors. METHODS: NSCLC cell lines were treated with HDAC inhibitors and analyzed for E-cadherin induction, growth inhibition and apoptosis. National Cancer Institute-H157 cells expressing ectopic E-cadherin were tested for tumorigenicity in murine xenografts. RESULTS: We found that treatment with MS-275, compared to vorinostat (SAHA), valproic acid or trichostatin A, was most effective in E-cadherin up-regulation and persistence in non-small cell lung cancers. As with other tumor types and HDAC inhibitors, MS 275 inhibited growth and induced apoptosis. Importantly, blocking E-cadherin induction by short hairpin RNA resulted in less inhibition by MS-275, implicating the epithelial to mesenchymal phenotype process as a contributing factor. In contrast to H460 and H661, H157 cells were resistant to E-cadherin up-regulation by HDAC inhibitors. However, E-cadherin was restored, in a synergistic manner, by combined knockdown of ZEB-1 and ZEB-2. In addition, H157 cells stably transfected with E-cadherin were markedly attenuated in their tumor forming ability. Lastly, combining MS-275 with the microtubule stabilizing agent, paclitaxel, or 17 (allylamino)-17-demethoxygeldanamycin, a heat shock protein 90 inhibitor, resulted in synergistic growth inhibition. Since MS-275 has no reported activity against HDAC6, which regulates both microtubule and heat shock protein 90 functions, other mechanisms of synergy are anticipated. CONCLUSIONS: These results support the role of ZEB proteins and HDAC inhibitors in the pathogenesis and treatment of lung cancer. PMID- 20009912 TI - Iatrogenic tracheoesophageal fistula. PMID- 20009913 TI - Antiangiogenesis induced tumor cavitation in lung cancer. PMID- 20009914 TI - Dramatic response to low-dose erlotinib of epidermal growth factor receptor mutation-positive recurrent non-small cell lung cancer after severe cutaneous toxicity. AB - Erlotinib is increasingly being used for the treatment of non-small cell lung cancer. The recommended dose is 150 mg/day and no efficacy data is available for lower doses. We describe a case of dramatic tumor response to 50 mg erlotinib in a patient with EGFR mutation positive NSCLC who developed a severe rash on full dose erlotinib. Rash is known to correlate with response and survival in patients treated with erlotinib. Our case suggests that in the presence of rash, dose reductions to "subtherapeutic" levels remain effective and may prevent unnecessary early treatment termination. PMID- 20009915 TI - Marked tumor response and fatal hemoptysis during radiation for lung cancer in a human immunodeficiency virus-positive patient taking nelfinavir. PMID- 20009916 TI - Tracheoesophageal fistula associated with bevacizumab 21 months after completion of radiation therapy. PMID- 20009917 TI - Abacavir-based therapy does not affect biological mechanisms associated with cardiovascular dysfunction. AB - OBJECTIVE: To assess the effects of initiating abacavir-containing therapy on plasma lipids and cardiovascular biomarkers. DESIGN: Sub-study of the BICOMBO study in which participants were randomized to switch their nucleoside backbone to either abacavir/lamivudine or tenofovir/emtricitabine. METHODS: We assessed 48 week changes in fasting lipids and several biomarkers including serum high sensitivity C-reactive protein (hsCRP), monocyte chemoattractant protein-1 (MCP 1), osteoprotegerin, interleukin (IL)-6, IL-10, tumor necrosis factor alpha (TNF alpha), intercellular adhesion molecule-1 (ICAM-1), vascular adhesion molecule-1 (VCAM-1), selectin E and P, adiponectin, insulin, and D-dimer in otherwise healthy, virologically suppressed HIV-infected patients randomly switched to abacavir/lamivudine or tenofovir/emtricitabine with no history of cardiovascular disease, no prior abacavir or tenofovir use, and no virological failure or AIDS during follow-up. RESULTS: Eighty (46 abacavir/lamivudine and 34 tenofovir/emtricitabine) patients were included. Baseline characteristics were similar between groups and between patients in the sub-study vs. those not. There were no significant differences in baseline lipids and markers between groups. Although total (6.5 vs. -6.7%, P < 0.0001) and low-density lipoprotein (LDL) (8.6 vs. -9.1%, P = 0.004) cholesterol increased significantly in the abacavir/lamivudine group relative to the tenofovir/emtricitabine group, we found no significant changes in the biomarkers: CRP (-3.9 vs. 0.0%), MCP-1 (5.9 vs. 4.0%), osteoprotegerin (5.1 vs. -2.8%), IL-6 (0.0 vs. 0.0%), IL-10 (0.0 vs. 0.0%), TNF-alpha (0.0 vs. 0.0%), ICAM-1 (6.6 vs. 5.2%), VCAM-1 (0.02 vs. -0.01%), selectin E (-0.4 vs. 7.8%), selectin P (4.6 vs. 12.6%), insulin (-2.5 vs. 8.8%), adiponectin (-2.2 vs. 15.4%), and D-dimer (0.0 vs. 0.0%) (P > or = 0.12 for all comparisons). CONCLUSION: Abacavir/lamivudine increased total and LDL cholesterol compared with tenofovir/emtricitabine, but it did not cause inflammation, endothelial dysfunction, insulin resistance, or hypercoagulability in virologically suppressed HIV-infected patients. PMID- 20009920 TI - Sensitivity of V75I HIV-1 reverse transcriptase mutant selected in vitro by acyclovir to anti-HIV drugs. AB - Trials of acyclovir for herpes simplex virus 2 infection in herpes simplex virus 2/HIV-1 coinfected patients not on antiretroviral therapy demonstrated a decrease in herpes simplex virus 2 and HIV-1 replication. Recent studies indicated that acyclovir has direct anti-HIV-1 activity and can select for the HIV-1 V75I reverse transcriptase variant in vitro. We show that the V75I variant has decreased sensitivity to some nucleoside analogs but an increased sensitivity to zidovudine, results that may guide selection of highly active antiretroviral therapy regimens in patients harboring this variant. PMID- 20009919 TI - Protease polymorphisms in HIV-1 subtype CRF01_AE represent selection by antiretroviral therapy and host immune pressure. AB - BACKGROUND: Most of our knowledge about how antiretrovirals and host immune responses influence the HIV-1 protease gene is derived from studies of subtype B virus. We investigated the effect of protease resistance-associated mutations (PRAMs) and population-based HLA haplotype frequencies on polymorphisms found in CRF01_AE pro. METHODS: We used all CRF01_AE protease sequences retrieved from the LANL database and obtained regional HLA frequencies from the dbMHC database. Polymorphisms and major PRAMs in the sequences were identified using the Stanford Resistance Database, and we performed phylogenetic and selection analyses using HyPhy. HLA binding affinities were estimated using the Immune Epitope Database and Analysis. RESULTS: Overall, 99% of CRF01_AE sequences had at least 1 polymorphism and 10% had at least 1 major PRAM. Three polymorphisms (L10 V, K20RMI and I62 V) were associated with the presence of a major PRAM (P < 0.05). Compared to the subtype B consensus, six additional polymorphisms (I13 V, E35D, M36I, R41K, H69K, L89M) were identified in the CRF01_AE consensus; all but L89M were located within epitopes recognized by HLA class I alleles. Of the predominant HLA haplotypes in the Asian regions of CRF01_AE origin, 80% were positively associated with the observed polymorphisms, and estimated HLA binding affinity was estimated to decrease 19-40 fold with the observed polymorphisms at positions 35, 36 and 41. CONCLUSION: Polymorphisms in CRF01_AE protease gene were common, and polymorphisms at residues 10, 20 and 62 most likely represent selection by use of protease inhibitors, whereas R41K and H69K were more likely attributable to recognition of epitopes by the HLA haplotypes of the host population. PMID- 20009918 TI - A genome-wide association study of carotid atherosclerosis in HIV-infected men. AB - BACKGROUND: The role of host genetics in the development of subclinical atherosclerosis in the context of HIV-infected persons who are being treated with highly active antiretroviral therapy (HAART) is not well understood. METHODS: The present genome-wide association study (GWAS) is based on 177 HIV-positive Caucasian males receiving HAART who participated in the Fat Redistribution and Metabolic Change in HIV Infection (FRAM) Study. Common and internal carotid intima-media thicknesses (cIMT) measured by B-mode ultrasound were used as a subclinical measure of atherosclerosis. Single nucleotide polymorphisms (SNPs) were assayed using the Illumina HumanCNV370-quad beadchip. Copy Number Variants (CNV) were inferred using a hidden Markov Model (PennCNV). Regression analyses were used to assess the association of common and internal cIMT with individual SNPs and CNVs, adjusting for age, duration of antiretroviral treatment, and principal components to account for potential population stratification. RESULTS: Two SNPs in tight linkage disequilibrium, rs2229116 (a missense, nonsynonymous polymorphism (IIe to Val)) and rs7177922, located in the ryanodine receptor (RYR3) gene on chromosome 15 were significantly associated with common cIMT (P value < 1.61 x 10). The RYR gene family has been known to play a role in the etiology of cardiovascular disease and has been shown to be regulated by HIV TAT protein. CONCLUSION: These results suggest that in the context of HIV infection and HAART, a functional SNP in a biologically plausible candidate gene, RYR3, is associated with increased common carotid IMT, which is a surrogate for atherosclerosis. PMID- 20009922 TI - Comparison of outcomes in young versus nonyoung patients with ST elevation myocardial infarction treated by primary angioplasty. AB - OBJECTIVES: We sought to determine in-hospital and intermediate-term outcomes of primary percutaneous coronary intervention (PCI) for ST elevation myocardial infarction (STEMI) in young adults. METHODS: We reviewed 2424 consecutive patients treated with primary angioplasty for acute MI; 465 were aged 45 or less (young group) and 1959 were 46-74 years of age (nonyoung group). Clinical characteristics, in-hospital and intermediate-term outcomes of primary PCI were analyzed. RESULTS: Compared with nonyoung patients, the young patients had significantly lower in-hospital and intermediate-term mortality (for in-hospital mortaliy: 5.4 vs. 1.2%, P<0.001; for intermediate-term mortality: 5 vs. 1.3%, P<0.001). By multivariate Cox regression analysis in all 2424 patients; cardiogenic shock, diabetes mellitus, anterior MI and unsuccessful procedure were independent predictors of both in-hospital and intermediate-term mortality whereas age [odds ratio (OR): 1.07, P<0.001], female sex (OR: 1.88, P = 0.04), MI history (OR: 3.05, P = 0.001) and multivessel disease (OR: 2.15, P = 0.01) were independent predictors of only intermediate-term mortality. The young group had lower unsuccessful procedure rates of primary PCI for STEMI (4.9 vs. 10.1%, P = 0.001). CONCLUSION: These results suggest that young adults who underwent primary PCI have favorable in-hospital and intermediate-term outcomes. Moreover, primary PCI for young adults with STEMI is safer, more feasible and effective than for a relatively older population. PMID- 20009921 TI - Dopamine is involved in the antidepressant-like effect of allopregnanolone in the forced swimming test in female rats. AB - Evidence from both animal and human studies suggests a role for dopamine in the therapeutic effect of antidepressant drugs. Consistently, dopamine receptor antagonists antagonize the effect of antidepressant drugs in different experimental models of depression. Neurosteroids, and in particular allopregnanolone, seem to be involved both in the pathophysiology of depression and in the mechanism of action of antidepressant drugs, and their role seems to be particularly important in the understanding of mood disturbances related to the different phases of the reproductive life in women. The aim of this study was to investigate the possible role of dopamine on the antidepressant-like effect of allopregnanolone in a model of depression. Thus, we examined (i) the behaviour of female Sprague-Dawley rats in the forced swimming test during estrus and diestrus and their response to allopregnanolone treatment (0.5, 1 and 2 mg/kg), and (ii) the effect of the dopamine D1-like and D2-like receptor antagonists SCH 23390 (0.01 and 0.025 mg/kg) and raclopride (0.05 and 0.2 mg/kg) on the antidepressant like effect of allopregnanolone (2 mg/kg) in the same experimental model. We failed to observe differences in depressive-like behaviour between estrous phases, and allopregnanolone administration in both estrus and diestrus resulted in an antidepressant-like effect consisting in an increase of swimming behaviour. The allopregnanolone effect was unaffected by a dose of the dopamine D1-like receptor antagonist SCH 23390 displaying a marked inhibitory effect on basal activity, while it was turned into a potentiation of the depressive-like behaviour of the forced swimming condition by treatment with the higher dose of raclopride. The present results indicate an involvement of dopamine transmission in the allopregnanolone antidepressant-like effect in the forced swimming model of depression, and suggest that this effect depends mainly on stimulation of dopamine D2-like receptors. PMID- 20009923 TI - Current challenges of stroke treatment. PMID- 20009924 TI - Intracerebral hemorrhage: getting ready for effective treatments. AB - PURPOSE OF REVIEW: Spontaneous intracerebral hemorrhage (ICH) is the most devastating type of stroke and a leading cause of disability and mortality in the United States and the rest of the world. The purpose of this article is to review recent advances in the management of spontaneous intracerebral hemorrhage. RECENT FINDINGS: Although no interventions have consistently shown an improvement of mortality or functional outcomes after ICH, results from multicenter prospective randomized controlled trials have shown that early hemostasis to prevent hematoma growth, removal of clot by surgical or minimally invasive interventions, clearance of intraventricular hemorrhage, and adequate blood pressure control for the optimization of cerebral perfusion pressure may constitute the most important therapeutic goals to ameliorate secondary neurological damage, decrease mortality, and improve functional outcomes after ICH. CONCLUSION: Several promising methods may be ready for routine clinical use in a few years to decrease disability and mortality from ICH. PMID- 20009925 TI - Optical coherence tomography of the retina: applications in neurology. AB - PURPOSE OF REVIEW: This review summarizes the mechanisms and recent developments of optical coherence tomography and its practical uses in neurology. The application of optical coherence tomography imaging of the retina in multiple sclerosis, neuromyelitis optica, Alzheimer disease, and Parkinson disease are reviewed. RECENT FINDINGS: Thinning of the peripapillary retinal nerve fibre layer has been detected in patients with optic neuritis, multiple sclerosis, neuromyelitis optica, Alzheimer disease, and Parkinson disease. However, the patterns of change differ in some aspects. SUMMARY: The findings indicate loss of retinal ganglion cells and may reflect degenerative change in the brain in these conditions. The retinal nerve fibre layer thickness may be used as a biological marker and may help to distinguish between optic neuritis associated with multiple sclerosis and optic neuritis in neuromyelitis optica. PMID- 20009926 TI - Antiangiogenic agents in the treatment of nonsmall cell lung cancer: reality and hope. AB - PURPOSE OF REVIEW: Angiogenesis is critical for tumor growth and progression. Strategies to inhibit angiogenesis have gained a strong foothold for the treatment of a variety of malignancies. This review will provide the relevant interventions targeted against specific angiogenesis pathways with or without known effective therapies for nonsmall cell lung cancer (NSCLC). RECENT FINDINGS: Bevacizumab, a mAb against the vascular endothelial growth factor, has been approved by the U.S. Food and Drug Administration for the treatment of patients with advanced stage nonsquamous NSCLC in combination with the carboplatin paclitaxel regimen. This has prompted the evaluation of a variety of novel agents for the treatment of NSCLC. Agents that inhibit the vascular endothelial growth factor receptor tyrosine kinase are currently under extensive investigation, but the initial results with combination strategies have not been encouraging. Identification of predictive biomarkers for antiangiogenic agents continues to be elusive and remains a major focus of ongoing research. Apart from vascular endothelial growth factor, other targets within the angiogenic pathway are also being evaluated in the clinical setting. SUMMARY: In this article, we review the recent data with antiangiogenic agents in NSCLC and their implications for clinical use and future research. PMID- 20009927 TI - Multimodality management of non-small cell lung cancer patients with brain metastases. AB - PURPOSE OF REVIEW: Brain metastases are an important cause of morbidity and mortality, and are the most common intracranial tumors in adults. The prognosis of patients with brain metastases is very poor. The increasing incidence of brain metastases is directly related to the improvements in the treatment of systemic disease. Commonly, brain metastases are discovered after the diagnosis of cancer, often after other systemic metastases have developed. In this review article, we present the standard treatment approach and discuss new directions. RECENT FINDINGS: The most widely used treatment for patients with brain metastases is whole-brain radiotherapy. Actually, surgery and radiotherapy remain the principal therapeutic interventions. In contrast, the benefit of chemotherapy has long been viewed with skepticism.In an effort to improve the therapeutic ratio, radiosensitizers are often used concurrently with external-beam radiation: motexafin gadolinium and efaproxaril.Novel anticancer agents are under clinical investigation. SUMMARY: The management of patients with brain metastases with non small cell lung cancer has improved over time, due to the development of new treatment options and a better knowledge of prognostic factors.In the next 5 years, the results of several ongoing multicenter randomized trials will become available to further define the role of various radiation sensitizers and chemotherapeutic agents in combination with stereotactic radiosurgery, whole brain radiation therapy, or both. PMID- 20009928 TI - Biological modulation of renal ischemia-reperfusion injury. AB - PURPOSE OF REVIEW: Biological modulation of renal ischemia-reperfusion injury holds the potential to reduce the incidence of early graft dysfunction and to safely expand the donor pool with kidneys that have suffered prolonged ischemic injury before organ recovery. RECENT FINDINGS: In the current review, we will discuss clinical studies that compare kidney transplant recipients with and without early graft dysfunction in order to elucidate the pathophysiology of ischemic acute allograft injury. We will specifically review the mechanisms leading to depression of the glomerular filtration rate and activation of the innate immune system in response to tissue injury. SUMMARY: We conclude that the pathophysiology of delayed graft function after kidney transplantation is complex and shares broad similarity with rodent models of ischemic acute kidney injury. Given the lack of specific therapies to prevent delayed graft function in transplant recipients, comprehensive efforts should be initiated to translate the promising findings obtained in small animal models into clinical interventions that attenuate ischemic acute kidney injury after transplantation. PMID- 20009929 TI - Pancreas transplantation. PMID- 20009930 TI - Pancreas vs. islet transplantation: a call on the future. AB - PURPOSE OF REVIEW: The aim of this article is to review recent reports on whole pancreas and islet cell transplantation. It focuses on 'what the call to the future looks like' for both therapies as treatment options for those type 1 diabetes patients who do not respond well to conventional therapy. RECENT FINDINGS: The major benefit of pancreas transplantation is the reversal of diabetes improvement of diabetes complications. Although the procedure requires major surgery and life-long immunosuppression, it remains the gold standard for a specific population of patients who suffer from type 1 diabetes and who do not respond to conventional therapy. Allogeneic islet transplantation is a promising alternative to pancreas transplantation, but patient outcomes remain less than optimal and significant progress is required in order for this procedure to be considered a reliable therapy. CONCLUSION: Several factors have to be taken into consideration before making the decision of which of these procedures would better suit a patient with type 1 diabetes. PMID- 20009931 TI - Complications after pancreas transplantation. AB - PURPOSE OF REVIEW: The history of transplantation of the pancreas, unlike that of transplantation of other abdominal organs, has largely been shaped by the associated surgical complications. After more than three decades of progress, surgical-technical pancreas graft failure rates have decreased to approximately 8%. The most recent developments in this area are systematically reviewed in this article. RECENT FINDINGS: Vascular graft thrombosis remains, by far, the most common cause of technical graft failure. Recent reports suggested that pancreas preservation with histidine-tryptophan-ketoglutarate solution (HTK) might be a risk factor for reperfusion pancreatitis, graft thrombosis and decreased short- and long-term graft survival. It remains unclear whether these results are, at least in part, related to HTK flush volumes and extended preservation (e.g.,>12 h). For selected thrombosed pancreas grafts, there has been renewed interest in pharmacological, interventional, and surgical salvage. For selected recipients with early pancreas graft thrombosis not amenable to a salvage intervention, transplant pancreatectomy in conjunction with immediate retransplantation has emerged as a viable option. For graft thrombosis prevention, the enhanced backtable pancreas vascular reconstruction techniques (e.g., gastroduodenal artery revascularization) proposed by some authors await more formal study. For prevention of native vascular complications in high-risk recipients, several technical modifications have been reported. Developments with respect to other surgical complications (wound infection, pancreatitis, leak, and bleeding) have been more incremental. CONCLUSION: Recent evidence underscores the importance of judicious donor and recipient selection and of optimization of preservation and surgical factors for excellent short- and long-term pancreas transplant outcomes. PMID- 20009932 TI - Pancreas transplantation in the United States: a review. AB - PURPOSE OF REVIEW: The goal of pancreas transplantation is to restore normoglycemia in patients with labile diabetes. The results of this procedure improved over the years, but, although pancreas transplantation is not considered experimental anymore, there is often reluctance to recommend this procedure because of the complexity, especially for solitary pancreas transplants. This article reviews the current status of pancreas transplantation. RECENT FINDINGS: Many improvements have been made in the surgical techniques and immunosuppressive regimens. The overall rate of technical problems decreased, yet immunologic graft loss is still a problem in solitary pancreas transplants. Careful donor selection significantly decreased the risk of graft failure and therefore improved patient survival. SUMMARY: With modern immunosuppressive protocols and careful donor selection, patient survival rates and pancreas transplant graft function can be further improved in all three pancreas transplant categories. PMID- 20009933 TI - Endothelial transcripts uncover a previously unknown phenotype: C4d-negative antibody-mediated rejection. AB - PURPOSE OF REVIEW: In the last decade, there has been a growing recognition of alloantibody responses in organ transplantation, but phenotypes related to antibody-mediated rejection (ABMR) remain incompletely defined. This article reviews recent molecular studies in kidney allograft tissues that decipher molecular burden and mechanisms of ABMR, leading to discovery of a new phenotype: 'C4d-negative ABMR'. RECENT FINDINGS: High endothelial gene expression in kidney transplant biopsies with anti-human leukocyte antigen alloantibody indicates active antibody-mediated damage and poor graft survival, defining a previously unknown group of C4d-negative ABMR. C4d-negative ABMR is characterized by high intragraft endothelial gene expression, alloantibodies, histology of chronic ABMR (less frequently acute ABMR), and poor outcomes. Thus, endothelial molecular phenotype in biopsies with circulating antibody detects degree of active graft injury, and many of these transcripts reflect endothelial activation. C4d negative ABMR is twice as common as C4d-positive ABMR. Recognition of this new phenotype reveals ABMR (C4d positive or negative) as the most common cause of late kidney transplant loss. SUMMARY: C4d staining, although very useful, is insensitive for detecting ABMR. Measuring endothelial gene expression in biopsies from kidneys with alloantibody is a sensitive and specific method to diagnose ABMR and predict graft outcomes. PMID- 20009935 TI - The patient with coronary stents and antiplatelet agents: what to do and how to deal? AB - The management of patients with recent coronary artery stents presenting for noncardiac surgery has become a major topic of interest and concern for all perioperative care givers. The present review will update recent reports and particularly new guidelines as well as recommendations. Based on the available literature, all experts recommend avoiding premature discontinuation of antiplatelet drug therapy if possible except for a few surgical procedures. Drug eluting stents obviously carry more risks than bare-metal stents. PMID- 20009936 TI - Dexamethasone with bupivacaine increases duration of analgesia in ultrasound guided interscalene brachial plexus blockade. AB - BACKGROUND AND OBJECTIVE: Dexamethasone has been shown to prolong the duration of postoperative analgesia when given as an adjunct for peripheral nerve blocks. However, it has not been evaluated when given in conjunction with bupivacaine and clonidine to provide blockade of the brachial plexus at the interscalene level. The purpose of this investigation was to determine whether the addition of dexamethasone to interscalene brachial plexus block would prolong the duration of sensory analgesia in a group of patients undergoing outpatient shoulder arthroscopy. METHODS: This prospective, randomized, double-blind investigation was performed on 88 individuals undergoing shoulder arthroscopy. Patients received interscalene brachial plexus block using 20 ml of bupivacaine 5 mg ml( 1) with 1: 200,000 epinephrine and clonidine 75 microg. Patients were randomly assigned to receive either dexamethasone 8 mg or 0.9% NaCl as an adjuvant to the mixture. After discharge, patients recorded pain scores and analgesic consumption in a diary and estimated the time at which they perceived that the sensory block from the interscalene brachial plexus block resolved. This was based on pain, recovery of sensation and strength in the arm. Variables measured included demographics, timed pain intensity measurements, postoperative analgesic consumption, duration of analgesia and patient satisfaction. RESULTS: Dexamethasone prolonged median sensory (1457 vs. 833 min, P < 0.0001) and motor (1374 vs. 827 min, P < 0.0001) blockade compared with the control. At 24 h, the dexamethasone group had lower median verbal analogue scale scores compared with control (3.0 vs. 6.0). At 48 h, the two groups had similar median pain scores (4.0 vs. 5.0, dexamethasone vs. control, respectively). The opioid requirement in oxycodone equivalency was lower in the dexamethasone group than in the control group for the first 24 h, and similar thereafter. Median patient satisfaction scores were not significantly different between the two groups at 48 h (9.5 vs. 8.0, dexamethasone vs. control, respectively). CONCLUSION: The addition of dexamethasone to a bupivacaine-epinephrine-clonidine interscalene block prolongs sensory block and reduces opioid use. PMID- 20009937 TI - Trends in cancer mortality in Brazil, 1980-2004. AB - Scanty information, limited to selected areas of the country, is available on cancer mortality in Brazil. Age-standardized (world population) mortality rates between 1980 and 2004, derived from the WHO database, were computed for all cancers and 24 major cancer sites in Brazil. Joinpoint regression analyses were used to identify the significant changes in trends and estimate annual percent change (APC) in rates. Total cancer mortality rates increased over the last decade in men (APC = 0.5) to reach 101.2/100 000, and in women (APC = 0.3) to reach 71.3/100 000. In men, upward trends were observed for cancers of the oral cavity and pharynx with a rate of 5.9/100 000 in 2000-2004, intestines (whose rate, however was low, i.e. 7.6), prostate (12.2), and leukemias (3.4). Male lung cancer increased until 1993 (APC = 1.39) and decreased thereafter (APC = -0.29), with a relatively low rate of 16.2/100 000 in 2000-2004. In women, there were steady upward trends for cancers of the lung (APC = 2.3), reaching 6.2/100 000 in 2000-2004, and leukemias (2.5). Breast cancer mortality leveled off at around 10/100 000 in the last decade, whereas declines were observed for cancers of the uterus, whose rate (8.3) however, remained comparatively high. Declines were observed for stomach cancer in both sexes, with rates of 11.1 in men and 4.6 in women. In conclusion, the key issues of cancer mortality in Brazil are the high rates of head and neck cancers in men and (cervix) uterine cancer in women, that is, in principle cancers that are largely avoidable through prevention, screening, and early diagnosis. PMID- 20009938 TI - Predictors of nonresponse to lactulose in patients with cirrhosis and hepatic encephalopathy. AB - BACKGROUND AND AIMS: Lactulose is commonly used in the treatment of hepatic encephalopathy (HE). However, all patients do not respond to lactulose. We evaluated predictors of nonresponse to lactulose in patients with cirrhosis and HE. PATIENTS AND METHODS: Consecutive cirrhotic patients with HE were enrolled. HE was diagnosed by West Haven criteria. Patients were treated with lactulose and correction of any associated precipitating factors. Nonresponse was defined if patient remained in HE even after 10 days of treatment or died while in HE. RESULTS: Of 300 patients with cirrhosis and HE, 231 (77%) patients met the inclusion criteria. The majority (95%) of the patients had Grade 2 or 3 HE. Of 231 patients, 180 (78%) responded to lactulose. Fifty-one (22%) did not respond to lactulose, 34 (15%) died without any improvement in HE and HE did not improve in 17 (7%) patients after 10 days of therapy. On comparing baseline parameters between nonresponders versus responders there was significant difference between baseline age (42.0+/-11.9 vs. 46.6+/-12.7 year, P=0.02), total leukocyte count (median, 9300 vs. 7300 cells/mm3, P=0.001), serum sodium level (129.9+/-6.2 vs. 133.7+/-7.1 mmol/l, P=0.001), model for end stage liver disease (MELD) score (22.9+/-3.8 vs. 19.9+/-4.2, P=0.001), mean arterial pressure (MAP, 77.9+/-10.0 vs. 86.3+/-8.7 mmHg, P=0.001), serum AST (median, 114 vs. 76 IU/l, P=0.01), serum ALT (median, 84 vs. 48.5 IU/l, P=0.001), spontaneous bacterial peritonitis [18 (35%) vs. 37 (21%), P=0.02] and hepatocellular carcinoma [HCC, 17 (33%) vs. 14 (7%), P=0.001]. On multivariate analysis baseline total leukocyte count, MELD, MAP, and HCC were independent predictors of nonresponse to lactulose (P=0.001). Combination of low MAP, high MELD, and presence of HCC had diagnostic accuracy of 81% in predicting nonresponse to lactulose. CONCLUSION: Of 78% patients with chronic liver disease with HE (majority with Grade 2 and 3) responded to lactulose. High baseline MELD, high total leukocyte count, low serum sodium, low MAP, and presence of hepatocellular carcinoma were predictors of nonresponse to lactulose. PMID- 20009939 TI - Ambulatory oesophageal pH monitoring: a comparison between antimony, ISFET, and glass pH electrodes. AB - BACKGROUND AND AIM: Ambulatory oesophageal pH-impedance monitoring is a widely used test to evaluate patients with reflux symptoms. Several types of pH electrodes are available: antimony, ion sensitive field effect transistor (ISFET), and glass electrodes. These pH electrodes have not been compared directly, and it is uncertain whether these different types of pH electrodes result in similar outcome. METHODS: In an in-vitro model the response time, sensitivity, and drift of an antimony, ISFET, and glass pH electrode were assessed simultaneously after calibration at 22 degrees C and at 37 degrees C. All measurements were performed at 37 degrees C and repeated five times with new catheters of each type. Fifteen patients with reflux symptoms underwent 24-h pH monitoring off PPI therapy using antimony, ISFET, and glass pH electrodes simultaneously. RESULTS: After calibration at 22 degrees C, pH electrodes had similar response times, sensitivity and drift. In contrast to glass electrodes, antimony electrodes performed less accurately after calibration at 37 degrees C than after calibration at 22 degrees C. Calibration temperature did not affect ISFET electrodes significantly. During in-vivo experiments, significant differences were found in acid exposure times derived from antimony (4.0+/-0.8%), ISFET (5.7+/-1.1%), and glass pH electrodes (9.0+/-1.7%). CONCLUSION: In vitro, antimony and glass pH electrodes are affected by different buffer components and temperature, respectively. In vivo, significant higher acid exposure times are obtained with glass electrodes compared with antimony and ISFET pH electrodes. ISFET electrodes produce stable in-vitro measurements and result in the most accurate in-vivo measurements of acid exposure time. PMID- 20009940 TI - Hepatic stellate cell activation and hepatic fibrosis in children with type 1 autoimmune hepatitis: an immunohistochemical study of paired liver biopsies before treatment and after clinical remission. AB - OBJECTIVES: The activation of hepatic stellate cells (HSC) is considered the most important event in hepatic fibrogenesis. The precise mechanism of this process is unknown in autoimmune hepatitis (AIH), and more evidence is needed on the evolution of fibrosis. The aim of this study was to assess these aspects in children with type 1 AIH. METHODS: We analyzed 16 liver biopsy samples from eight patients, paired before treatment and after clinical remission, performed an immunohistochemical study with anti-alpha actin smooth muscle antibody and graded fibrosis and inflammation on a scale of 0-4 (Batts and Ludwig scoring system). RESULTS: There was no significant reduction in fibrosis scores after 24+/-18 months (2.5+/-0.93 vs. 2.0+/-0.53, P=0.2012). There was an important decrease in inflammation: portal (2.6+/-0.74 vs. 1.3+/-0.89, P=0.0277), periportal/periseptal (3.0+/-0.76 vs. 1.4+/-1.06, P=0.0277), and lobular (2.8+/-1.04 vs. 0.9+/-0.99, P=0.0179). Anti-alpha actin smooth muscle antibodies were expressed in the HSC of the initial biopsies (3491.93+/-2051.48 mum), showing a significant reduction after remission (377.91+/-439.47 microm) (P=0.0117). CONCLUSION: HSC activation was demonstrated in the AIH of children. The reduction of this activation after clinical remission, which may precede a decrease in fibrosis, opens important perspectives in the follow-up of AIH. PMID- 20009941 TI - Wavefront-guided ablation retreatment using Iris registration. AB - PURPOSE: To evaluate the efficacy, predictability, safety, and intraoperative and postoperative complications of laser in situ keratomileusis (LASIK) retreatment in myopic eyes using wavefront-guided ablation with iris registration (IR). METHODS: Retrospective analysis was used to evaluate wavefront-guided retreatment with IR in a consecutive cohort of 77 eyes (57 patients) after LASIK. The eyes were divided into two groups: no previous retreatment group (group 1) (n = 63) and previous LASIK retreatment group (group 2) (n = 14). The primary outcome variables assessed postoperatively at 1, 3, and 6 months were uncorrected visual acuity (UCVA), best-corrected visual acuity (BCVA), and pre- and postretreatment changes in manifest refraction. RESULTS: The mean preretreatment spherical equivalent in group 1 was reduced from -0.5 +/- 1.0 diopter (D) (range -3 to 2.4) to 0.06 +/- 0.3 (range -0.9 to 0.6) (P < 0.002) at 6 months. In group 2, the mean preretreatment spherical equivalent was reduced from -0.9 +/- 1.24D (range -3.1 to -0.5) to 0.04 +/- 0.5 (range -1.0 to 1.1) (P < 0.049) at 6 months. At 6 months, UCVA was 20/20 or better in 92% in group 1 and 64% in group 2, of patients, respectively. No eyes lost more than one line of BCVA in group 1 and one eye (7%) lost two lines of BCVA in group 2. CONCLUSION: Wavefront-guided LASIK retreatment with IR after LASIK is an effective, predictable, and safe procedure in cases requiring a single retreatment. In contrast, eyes with previous retreatments showed less predictability and lower percentage of eyes with postoperative 20/20 UCVA. PMID- 20009942 TI - Measurement of the refractive index of soft contact lenses during wear. AB - PURPOSE: To determine whether the refractive index (RI) of a soft contact lens can be evaluated using refractometry while the lens remains on the eye and to compare this with more traditional ex vivo RI measurements. METHODS: A slitlamp apparatus was modified to incorporate a customized Atago hand refractometer. With a double-masked study design, nine adapted symptomatic soft contact lens wearers wore a contact lens in each eye (lotrafilcon B and etafilcon A) in a randomized order. In vivo RI was determined from the relative Brix scale measurements immediately after lens insertion and after 1 and 10 hr of lens wear. Ex vivo refractometry was performed after 10 hr of lens wear for comparison. Means +/- standard errors of the means are reported. RESULTS: In vivo RI values at baseline were 1.422 +/- 0.0004 (lotrafilcon B) and 1.405 +/- 0.0021 (etafilcon A); after 1 hr of lens wear, values were 1.423 +/- 0.0006 and 1.408 +/- 0.0007, respectively; and after 10 hr of lens wear, values were 1.424 +/- 0.0004 and 1.411 +/- 0.0010, respectively. Ex vivo RI values at the end of the 10 hr wearing period were 1.424 +/- 0.0003 (lotrafilcon B) and 1.412 +/- 0.0017 (etafilcon A). The change in in vivo RI across the day was statistically significant for the etafilcon A lens (repeated-measures analysis of variance, P<0.01) but not for the lotrafilcon B lens (P>0.05). CONCLUSIONS: This novel adaptation of refractometry was able to measure the RI of soft contact lenses during wear (without lens removal). End of day RI measurements using in vivo and ex vivo refractometry were comparable with each other. Future work is required to determine whether this in vivo method can improve our understanding of the relationships between soft contact lens RI, hydration, on-eye lens performance, and symptomology. PMID- 20009943 TI - Differences in the daily symptoms associated with the silicone hydrogel contact lens wear. AB - OBJECTIVE: To determine the differences in symptoms associated with daily wear of silicone hydrogel (lotrafilcon B) contact lenses (CLs) after CL insertion and at the end of wear and their relationship with environmental factors and multipurpose solutions (Solo-care Aqua, CIBA Vision, Duluth, GA and Hidro Health, Disop, Spain). METHODS: Fifty-four patients were fitted with lotrafilcon B CLs. Two multipurpose solutions were randomly assigned and prescribed in a double masked crossover study with three visits. After each solution was used, two questionnaires were conducted, including a survey produced by us and Contact Lens Dry Eye Questionnaire. Our questionnaire included 10 items addressing discomfort, blurry vision, lens-handling problems, dryness, redness, tearing, burning, itching, discharge, and dissatisfaction. Patients were assigned scores from 0 (without symptoms) to 10 (symptoms unbearable) at two different times (after CL insertion and at the end of wear) and in two environments (outdoors and adverse environments). RESULTS: The average duration of CL wear was 8.32 +/- 2.27 hr/day and 5.85 +/- 1.30 days/week. All symptoms became worse (P < 0.05 Wilcoxon test, except lens handling) at the end of CL wear in all visits. The results were independent of the multipurpose solution used (P > 0.05 Friedman test), except for tearing, which showed statistical differences between visits (P = 0.03 Friedman test). Contact Lens Dry Eye Questionnaire showed increased dry eye symptoms at the end of wear (P < 0.05 Friedman test). CONCLUSIONS: Daily wear symptoms associated with lotrafilcon B CL increase with the time of wear. This increase in symptoms is nonrelated with the multipurpose solutions compared in this study. PMID- 20009944 TI - Translocation of malpositioned posterior chamber intraocular lens from anterior to posterior chamber along with fibrin glue-assisted transscleral fixation. AB - PURPOSE: A posterior chamber intraocular lens (PCIOL) wrongly placed in anterior chamber (AC) can lead to progressive endothelial loss and damage to angle structures. We describe a sutureless, closed-chamber translocation technique to maneuver a PCIOL from AC into the PC and fixate it transsclerally with fibrin glue in presence of a large posterior capsular rent. METHODS: This is a prospective interventional case series. The participants were three cases of PCIOL in AC with high intraocular pressure and poor uncorrected visual acuity who underwent the translocation surgery successfully. RESULTS: All cases showed improvement in uncorrected visual acuity, with lowered intraocular pressure and no loss in best spectacle-corrected visual acuity till 6 months follow-up. CONCLUSIONS: This technique prevents a large incision, does not require a special IOL as in transscleral suture fixation, and results in a low endothelial cell loss. PMID- 20009945 TI - Scleral lenses in the management of keratoconus. AB - PURPOSE: To describe the use of Jupiter scleral lenses (Medlens Innovations, Front Royal, VA; and Essilor Contact Lenses, Inc., Dallas, TX) in the management of keratoconus. METHODS: We performed a single-center retrospective chart review of our initial 32 patients with keratoconus evaluated for scleral lens wear. All patients were referred for scleral lens evaluation after exhausting other nonsurgical options for visual correction. Diagnostic lenses were used in the initial fitting process. If adequate fit could not be achieved with standard lenses, custom lenses were designed in consultation with the manufacturers' specialists. The following measures were evaluated for each patient: ability to tolerate and handle lenses, visual acuity with scleral lenses, number of lenses, and visits needed to complete the fitting process. RESULTS: Fifty-two eyes of 32 patients were evaluated for scleral lens wear. Of these, 12 patients (20 eyes) decided not to pursue scleral lens wear after initial evaluation. One patient (2 eyes) abandoned the fitting process after cataract surgery. The remaining 19 patients (30 eyes) were fit successfully. The average number of lenses ordered per eye was 1.5. The fitting process required an average of 2.8 visits. Standard lenses were prescribed for 23 eyes, and custom designs were needed for 7 eyes. Median best-corrected visual acuity improved from 20/40 (mean, 20/76) before scleral lens fitting to 20/20 (mean, 20/30) after fitting. Follow-up ranged from 3 to 32 months. CONCLUSIONS: Jupiter scleral lenses provide acceptable visual acuity and comfort in patients with keratoconus. The availability of diagnostic lenses facilitates the fitting process. PMID- 20009946 TI - Ophthalmologist perceptions regarding treatment of moderate-to-severe dry eye: results of a physician survey. AB - PURPOSE: To understand ophthalmologists' current perceptions and treatment of patients with moderate-to-severe dry eye disease (DED). SETTING: Online survey. METHODS: The online survey was sent to 7,882 ophthalmologists, including 51 corneal specialists, throughout the United States from October 9 to 21, 2008. The response rate was 3.1% (n = 245), typical for this type of survey. Only ophthalmologists who treated four or more moderate-to-severe DED patients per month (235 of 245 [96%]) were asked to complete the survey. RESULTS: Ninety-four percent of respondents agreed that more treatment options are needed for moderate to-severe DED. Corneal specialists were more likely to strongly agree (63%) than general ophthalmologists (54%). Only 33% overall felt that current therapies were extremely or very effective for moderate DED and only 5% for severe disease. Ninety-two percent agreed that multiple therapeutic agents are needed to manage moderate-to-severe DED. The respondents prescribed or recommended a mean of 3.2 different treatments (standard deviation = 1.2) for moderate DED patients over the course of a year and 4.9 (standard deviation = 2.2) for patients with severe DED. The most highly ranked goals for treatment of moderate-to-severe DED patients were maintaining and protecting the ocular surface (ranked 1 or 2 x 74%) and lubricating and hydrating the ocular surface (ranked 1 or 2 x 67%). Corneal specialists ranked maintaining and protecting the ocular surface even more highly (ranked 1 or 2 x 82%). CONCLUSIONS: Results reflected the difficulty of treating moderate-to-severe DED, the importance of using multiple treatment approaches, the limitations of current treatment options, and the need for additional treatment options. PMID- 20009947 TI - Disinfection efficacy and encystment rate of soft contact lens multipurpose solutions against Acanthamoeba. AB - OBJECTIVES: To assess the ability of commercial and experimental soft contact lens multipurpose solutions (MPS) to promote Acanthamoeba trophozoite encystment and their biocidal efficacy against Acanthamoeba trophozoites and cysts. The effects on encystment and disinfection efficacy by the incorporation of propylene glycol (PG) in the formulation of MPS were also investigated. METHODS: Acanthamoeba trophozoites (Acanthamoeba castellanii ATCC 50730 and Acanthamoeba polyphaga CCAP 1501/3G) were inoculated into MeniCare Soft, MeniCare Soft (-PG), Epica Cold II, OptiFree Plus, and Rohto C cube Softone-Moist MPS, and the percentage of encystment induced by each solution was determined after 24 hr. In addition, Acanthamoeba trophozoites and cysts (A. castellanii ATCC 50730 and A. polyphaga CCAP 1501/3G and Ros) were also inoculated into each of the five MPS, and their log reduction determined after 0, 1, 4, 6, and 24 hr of incubation using stand-alone assays. RESULTS: Significantly higher encystment rates were found with Epica Cold II for A. polyphaga CCAP 1501/3G and Rohto C cube Softone Moist for A. polyphaga and A. castellanii compared with the other MPS assessed (P<0.05). MeniCare Soft, MeniCare Soft (-PG), and Opti-Free Plus produced little or no encystment, with mean encystment values ranging from 0.0% to 2.0%. A significantly higher disinfection efficacy was found with MeniCare Soft, MeniCare Soft (-PG), and Epica Cold II compared with Opti-Free Plus and Rohto C cube Softone-Moist (P<0.05). CONCLUSIONS: Significant differences in encystment rate and disinfection efficacy between MPS were found. The presence of PG in the formulation of MeniCare Soft did not affect the disinfection efficacy or the encystment rate. The latter indicates that other factors play a role in the induction of Acanthamoeba encystment after inoculation into MPS. PMID- 20009948 TI - In vitro contact angle analysis and physical properties of blister pack solutions of daily disposable contact lenses. AB - OBJECTIVES: The purpose of this study was to measure the advancing and receding contact angles (CAs) of five daily disposable (DD) lenses and the osmolality, surface tension (ST), and pH of each blister pack solution. METHODS: The advancing and receding CAs were measured directly out of the blister pack for five DD lenses: omafilcon A (CooperVision), nelfilcon A (CIBA Vision), modified nelfilcon A (CIBA Vision), etafilcon A (Johnson & Johnson), and narafilcon A (Johnson & Johnson). Advancing CAs were measured using sessile drop and Wilhelmy balance methods. Receding CAs were measured using the Wilhelmy balance method. ST, pH, and osmolality were measured for each blister pack solution from all the DD lenses. RESULTS: The advancing CAs for the nelfilcon A lenses were statistically lower (P<0.05) than the advancing CAs of the other three lenses. The receding CAs for etafilcon A were statistically lower (P<0.05) than the receding CAs for the other four lenses. The pH of all the blister pack solutions was relatively neutral (7.01-7.43). The ST of the blister pack solution for the modified nelfilcon A was significantly lower (P<0.05) than the ST of all other blister pack solutions. The osmolality of the blister pack solutions for the etafilcon A and narafilcon A lenses were significantly higher (P<0.05) than the other blister pack solutions. CONCLUSIONS: The modified nelfilcon A lens had low advancing CAs and low receding CAs showing minimal hysteresis. The blister pack solution for the modified nelfilcon A lens had the lowest ST, a low osmolality, and a neutral pH. PMID- 20009949 TI - CD4+ T-cell restoration after 48 weeks in the maraviroc treatment-experienced trials MOTIVATE 1 and 2. AB - OBJECTIVES: To determine factors associated with CD4 responses to maraviroc (MVC) containing regimens in treatment-experienced patients. METHODS: Forty-eight-week data from MOTIVATE 1 and 2 was used to assess MVC once or twice daily versus placebo (PBO), each with optimized background therapy (N = 1047). A repeated measures model evaluated longitudinal CD4 changes, multivariate linear regression evaluated predictors of week 48 increases, and Cox proportional hazard modeling evaluated time to category C events. RESULTS: Median CD4 increases were greater on MVC once or twice daily than PBO (92, 103, and 24 cells/mm3, respectively; P < 0.05), and the difference remained significant among patients achieving less than 50 HIV-1 RNA copies/mL (126, 125, and 96 cells/mm3; P < 0.05) or when adjusted for other predictors of CD4 increase including change in HIV-1 RNA. Time to a category C event was longer on MVC; in multivariate models, higher on-treatment CD4 count, but not MVC treatment, was protective against new events (hazard ratio 0.8 per +25 cells/mm3; 95% confidence interval 0.78-0.87). CONCLUSIONS: MOTIVATE patients receiving MVC had larger CD4+ T-cell increases than those receiving PBO, even after adjusting for the greater virologic potency of MVC-containing regimens. This additional CD4 response was associated with a longer time to the development of AIDS-defining events on MVC. PMID- 20009950 TI - Clinical practice: diagnosis and evaluation of dyspepsia. AB - The main issue regarding the approach to the patient with uninvestigated dyspepsia is whether the symptoms are the result of an important clinical illness, which then determines the appropriate management strategy for the treatment of the symptoms. An initial trial of empiric antisecretory drugs is recommended for those without Helicobacter pylori infection and no alarm symptoms, whereas H. pylori eradication is recommended for those with an active H. pylori infection. Treatment expectations for H. pylori infections should theoretically be similar to other common infectious diseases. In most regions, clarithromycin resistance has undermined traditional triple therapy so that it is no longer a suitable choice as an empiric therapy. Four drug therapies, such as sequential, concomitant, and bismuth-quadruple therapy are generally still acceptable choices as empiric therapies. Posteradication testing is highly recommended to provide early identification of otherwise unrecognized increasing antimicrobial resistance. However, despite the ability to successfully cure H. pylori infections, a symptomatic response can be expected in only a minority of those with dyspepsia not associated with ulcers (so called nonulcer dyspepsia). Overall, from the patients stand point, symptomatic relief is often difficult to achieve and physicians must rely on reassurance along with empiric and individualized care. PMID- 20009951 TI - International outsourcing of medical research by high-income countries: changes from 1995 to 2005. AB - BACKGROUND: Medical research outsourcing provides a financial benefit to those conducting research and financial incentives to the developing countries hosting the research. Little is known about how frequently outsourcing occurs or the type of research that is outsourced. METHODS: To document changes in medical research outsourcing over a 10-year period, we conducted a cross-sectional comparison of 3 medical journals: Lancet, The New England Journal of Medicine, and JAMA: The Journal of the American Medical Association in the last 6 months of 1995 and 2005. The main outcome measure was the 10-year change in proportion of studies including patients from low-income countries. FINDINGS: We reviewed 598 articles. During the 10-year period, the proportion of first authors from low-income countries increased from 3% to 6% (P = 0.21), whereas studies with participants from low-income countries increased from 8% to 22% (P = < 0.001). In 2005, compared with studies conducted exclusively in high-income countries, those including participants from low-income countries were more likely to be randomized trials (55% vs 35%, P = 0.004), to study medications (65% vs 34%, P < 0.001), to be funded by pharmaceutical companies (33% vs 21%, P = 0.05), and to involve pediatric populations (29% vs 8%, P < 0.001). INTERPRETATION: Outsourcing of medical research seems to be increasing. Additional studies are required to know if subjects from low-income countries are being adequately protected. PMID- 20009952 TI - Screening and evaluation of study subjects in patient-oriented research. AB - This article describes practical aspects of screening and evaluation of potential study subjects for patient-oriented research. Screening and evaluation comprise the crucial initial steps in the process of any patient-oriented research study. Appropriate infrastructure, operations, and documentation of the procedures needed to efficiently and effectively conduct screening and evaluation are described. A stepwise approach for screening and evaluating potential study subjects for determination of study eligibility is described. The design and conduct of screening and evaluation procedures should be protocol driven and compliant with regulatory requirements. PMID- 20009953 TI - Creation and use of a database in clinical and translational research. AB - Often data collection for clinical studies is an afterthought. The results of such an approach are incomplete or confusing data that can, as a worst case, result in scrapping and restarting the entire study. We discuss the planning process for data collection and storage to include encounter form development; data flow and capture; data checking, verification, and validation; advantage of relational databases over spreadsheets; data security; and aspects of a complete data system. PMID- 20009954 TI - Randomized clinical trials in stroke research. AB - A randomized clinical trial is widely regarded as the most rigorous study design to determine the efficacy of intervention because spurious causality and bias associated with other experimental designs can be avoided. The purpose of this article is to provide clinicians and clinical researchers the types of randomized clinical trials used in stroke studies and to discuss the advantages and the limitations for each type of randomized stroke clinical trials. PMID- 20009955 TI - Emotional stress and tako-tsubo cardiomyopathy: observations on 2 distinct clinical phenotypes. AB - BACKGROUND: Tako-tsubo syndrome is a transient cardiomyopathy usually precipitated by an acute emotional or physiological stress. Our study objectives were to review and analyze the impact of emotional stress on clinical variables, echocardiographic characteristics, and short-term outcomes in patients with tako tsubo syndrome. METHODS: Retrospective chart review. RESULTS: Eleven patients presented with tako-tsubo syndrome (1 man and 10 women) during the 8-year period (January 2000 to January 2008). The patients were split into 2 groups, defined by presenting either after emotional stress (6/11, 54.5%) or after idiopathic/physical stress (5/11, 45.5%). The mean age was 53.8 (12.6) years. The mean peak troponin T level was 0.54 ng/mL (range, 0.03-2.06 ng/mL), and the mean left ventricular end-diastolic pressure was 15.8 (8.1) mm Hg. Emotional stress was associated with younger age (P = 0.024), a lower left ventricular end diastolic pressure (P < 0.05), more ST segment changes on electrocardiogram (66.7% vs 40%), and a higher ejection fraction (P = 0.012). The patients in the idiopathic/physical stress group required more frequent hemodynamic support. CONCLUSIONS: We conclude that clinical, echocardiographic, and outcome characteristics can differentiate tako-tsubo patients presenting with emotional stress from those with idiopathic/physical stress into 2 distinct clinical phenotypes. Clinicians should monitor idiopathic/physically stressed tako-tsubo patients carefully for hemodynamic compromise. PMID- 20009956 TI - Ezetimibe effect on bone mineral density and markers of bone formation and resorption. AB - BACKGROUND: Ezetimibe, as a lipid-lowering agent, inhibits the intestinal absorption of cholesterol and decreases low-density lipoprotein cholesterol (LDL C) level in serum. It also up-regulates hepatic cholesterol biosynthesis and, by contrast to statins, increases serum mevalonate levels. Statins and biphosphonates decrease osteoclastic activity through the same mechanisms by inhibiting the mevalonate pathway. We therefore tested the effect of ezetimibe on bone turnover in hypercholesterolemic patients. SUBJECT AND METHODS: In an open label clinical trial, 54 hypercholesterolemic patients included in the study underwent 12 months of treatment with ezetimibe at a dosage of 10 mg/d. Before and after the 1-year ezetimibe treatment, bone mineral density (BMD) was measured by dual-energy x-ray absorptiometry and serum samples taken for measurements of levels of total cholesterol (TC), triglyceride, high-density lipoprotein cholesterol and LDL-C, serum calcium (Ca), serum phosphate, total and bone alkaline phosphatases (ALPs), and carboxyterminal fragment of type 1 collagen in the serum. RESULTS: The hypercholesterolemic patients showed a significant reduction with respect to baseline TC and LDL-C serum levels: 20% for TC (270.18 [38.58]-214.46 [38] mg/dL) and 24% for LDL-C (189.57 [38.58]-144 [32.05] mg/dL). Biochemical markers of both bone formations (total ALP level, 65.50 [21.33]-66.27 [21.017] IU/L and bone ALP level, 55.93 [7.92]-56.25 [7.49] IU/L) and bone resorption (beta-CTx, 0.44 [0.24]-0.46 [0.21] ng/mL) increased but did not show any significant change for the whole study period. At the end of 1 year, both BMD lumbar spine (0.90 [0.12]-0.89 [0.08] g/cm) and BMD-total femur (0.93 [0.12]-0.92 [0.12] g/cm) showed a negative trend but without reaching statistical significance. CONCLUSIONS: Our study results showed a negative trend but did not demonstrate statistically significant changes of BMD and metabolic markers with the treatment of ezetimibe. PMID- 20009957 TI - A novel treatment for metastatic melanoma with intralesional rose bengal and radiotherapy: a case series. AB - Three patients with metastatic melanoma were treated with intralesional rose bengal followed by external beam radiotherapy. In all cases patients had an impressive response without significant increase in acute radiation reaction. PMID- 20009958 TI - The Acupuncture on Hot Flashes Among Menopausal Women study: observational follow up results at 6 and 12 months. AB - OBJECTIVE: The previously published Acupuncture on Hot Flashes Among Menopausal Women study compared the effectiveness of individualized acupuncture treatment plus self-care versus self-care alone on hot flashes and health-related quality of life in postmenopausal women. This article reports on the observational follow up results at 6 and 12 months. METHODS: The Acupuncture on Hot Flashes Among Menopausal Women study was a pragmatic, multicenter randomized controlled trial with two parallel arms, conducted in 2006 to 2007. The 267 participants were postmenopausal women experiencing, on average, 12.6 hot flashes per 24 h. The acupuncture group received 10 individualized acupuncture treatments during 12 weeks and advice on self-care, whereas the control group received only advice on self-care. Hot flash frequency and intensity (0-10 scale) and hours of sleep per night were registered in a diary. Health-related quality of life was assessed by the Women's Health Questionnaire. RESULTS: From baseline to 6 months, the mean reduction in hot flash frequency per 24 hours was 5.3 in the acupuncture group and 5.0 in the control group, a nonsignificant difference of 0.3. At 12 months, the mean reduction in hot flash frequency was 6.0 in the acupuncture group and 5.8 in the control group, a nonsignificant difference of 0.2. Differences in quality-of-life scores were not statistically significant at 6 and 12 months. CONCLUSION: The statistically significant differences between the study groups found at 12 weeks were no longer present at 6 and 12 months. Acupuncture can contribute to a more rapid reduction in vasomotor symptoms and increase in health related quality of life in postmenopausal women but probably has no long-term effects. PMID- 20009959 TI - Extract and fraction from Ulmus wallichiana Planchon promote peak bone achievement and have a nonestrogenic osteoprotective effect. AB - OBJECTIVE: This study aimed to determine the skeletal effects of total ethanolic extract (TEE) and its butanolic fraction (BF) from the stem-bark of Ulmus wallichiana, which is rich in C-glycosylated flavonoids, in growing rats (for peak bone [PB] achievement) and in ovariectomized (OVx) rats (for menopausal bone loss). METHODS: TEE (750 mg kg(-1) d(-1)) and BF (50 mg kg(-1) d(-1)) were given orally for 10 weeks to weaning female Sprague-Dawley rats and for 12 weeks to adult OVx rats of the same strain, respectively. In studies with OVx rats, sham operated + vehicle, OVx + 17beta-estradiol, and OVx + vehicle groups served as various controls. Bone mineral density (BMD), biomechanical strength, bone histology, formations of osteoprogenitor cells, osteoid formation, and bone turnover/resorption markers were studied. Bioactive marker compounds in TEE and BF were analyzed by high-performance liquid chromatography. One-way analysis of variance was used to test significance of effects. RESULTS: In growing rats, both TEE and BF increased BMD, bone strength, and bone formation rate, suggesting higher PB achievement. OVx rats treated with either TEE or BF exhibited increased BMD at various anatomical positions and improved bone strength and trabecular architecture compared with the OVx + vehicle group. Serum osteocalcin and urinary type 1 collagen degradation product levels in OVx rats treated with either TEE or BF were significantly lower than those of the OVx + vehicle group. Neither TEE nor BF exhibited uterine estrogenicity. Analysis of marker compounds revealed significant enrichment of two bioactive markers in BF over TEE. CONCLUSIONS: Derived from U wallichiana, BF at much a lower dose than TEE was effective in PB achievement and prevention of OVx-induced bone loss. PMID- 20009960 TI - Association between vasomotor hot flashes and heart rate variability in recently postmenopausal women. AB - OBJECTIVE: The aim of this study was to investigate whether cardiovascular autonomic reactivity and risk profile are associated with the frequency and severity of hot flashes in recently postmenopausal women. METHODS: A total of 150 postmenopausal women with varying degrees of severity of hot flashes (none, mild, moderate, or severe) underwent 24-hour electrocardiographic recording. The function of the autonomic nervous system was assessed via heart rate variability in time and frequency domains. The effects of hot flashes on cardiac autonomic function were studied by assessing heart rate variability in the presence and absence of symptoms. RESULTS: There were no differences in mean heart rate, heart rate extremes, or total number of ectopic beats between women without and women with mild, moderate, or severe hot flashes. However, most women (14/17, 82%) with frequent ventricular ectopic beats and all women with ventricular runs belonged to the symptomatic groups. Although there were no differences in 24-hour or nighttime heart rate variability between the study groups, the very-low-frequency spectral component of heart rate variability increased by 72% (P < 0.001) during the hot flash period compared with the control period and was accompanied by an increase in heart rate (3%; P < 0.001). CONCLUSIONS: Cardiovascular risk markers based on heart rate variability failed to show an association with the frequency and severity of hot flashes in recently postmenopausal women. However, during a hot flash episode, there were signs of altered autonomic control of heart rate, which may be involved in the regulatory mechanisms of hot flashes. PMID- 20009962 TI - Oscillation development for neurotransmitter-related genes in the mouse striatum. AB - The aims of this study were to determine (i) whether striatal neuropeptides (dynorphin, enkephalin 1, substance P, cholecystokinin) and dopamine receptors 1 and 2 (D1r and D2r) are regulated by the molecular clock; and (ii) when their oscillations start after birth. Twenty-four-hour mRNA oscillations of these genes were evaluated in the mouse striatum at early postnatal stage (postnatal day 3), preweaning stage (postnatal day 14), and adult (postnatal day 60). At P3, no daily oscillations were observed. A significant time effect was present for D2r, dynorphin, and enkephalin 1 at P14, and for all genes except D1r, at P60. In conclusion, circadian expression of these neurotransmitter-related genes develops in the mouse striatum after birth gradually. PMID- 20009963 TI - Utility of 18F-FDG PET/CT for staging NK/T-cell lymphomas. AB - PURPOSE: Extranodal natural killer (NK)/T-cell lymphoma is a rare neoplasm and limited data has reported regarding the utilization of fluorine-18, fluorodeoxyglucose positron emission tomography/computed tomography (F-FDG PET/CT) in this disease. The aim of this study was to assess the role of F-FDG PET/CT in the staging of NK/T-cell lymphomas. PATIENTS AND METHODS: Thirteen newly diagnosed and two recurrent patients with NK/T-cell lymphoma who received F FDG PET/CT were studied. The lesion with intense F-FDG uptake was suggested as the positive and was measured using maximal standardized uptake values. The results of PET/CT were compared with the conventional staging examinations. RESULTS: F-FDG PET/CT detected nasal or extranasal lymphoma lesions in at least one site in all of the 15 patients. There was no significant difference of F-FDG uptake in lesions between patients with stage I-II disease and those with stage III-IV disease (maximal standardized uptake values 8.44+/-5.56 vs. 10.32+/-7.80; t=0.757, P>0.05). In two patients with an indeterminate diagnosis, the diagnosis of NK/T-cell lymphomas was established by biopsy guided by PET/CT and the status of stage IV was correctly identified. In 13 patients with definite diagnosis, the stage of disease was changed in six patients on the basis of F-FDG PET/CT. Two patients were down staged, and four patients upstaged. CONCLUSION: The lesions of the NK/T-cell lymphoma are F-FDG avid and PET/CT seems to be useful in the staging of this disease. PMID- 20009961 TI - Predictors of breast discomfort among women initiating menopausal hormone therapy. AB - OBJECTIVE: The aim of this study was to analyze the determinants of breast discomfort among postmenopausal women initiating menopausal hormone therapy (HT). METHODS: We analyzed questionnaire, anthropometric, and serum estrone data from the Postmenopausal Estrogen/Progestin Interventions (PEPI) Trial, a randomized trial comparing placebo, conjugated equine estrogens (CEE) alone, or CEE with a progestogen (continuous or cyclical medroxyprogesterone acetate or cyclical micronized progesterone) among postmenopausal women. HT users could join the PEPI Trial after stopping HT for 2 months. We modeled the relation between smoking, body weight, alcohol consumption, age, quitting HT for the PEPI Trial, physical activity, and alpha-tocopherol consumption and new-onset breast discomfort at the 12-month follow-up among 662 participants without baseline breast discomfort. RESULTS: The associations of new-onset breast discomfort with weight and with strenuous exercise varied by treatment assignment. Among women assigned to CEE + progestogen, strenuous exercise was associated with a 49% lower odds of new-onset breast discomfort (odds ratio, 0.51; 95% CI, 0.29-0.89; P = 0.02), whereas among women assigned to placebo or CEE alone, strenuous exercise was not significantly associated with new-onset breast discomfort. Surprisingly, among women taking CEE alone, each kilogram higher weight was associated with a 6% lower odds of new onset breast discomfort (P = 0.04), whereas among women taking placebo, the association was in the opposite direction (P = 0.04). Adjustment for estrone level had negligible effects on odds ratios. alpha-Tocopherol intake, age, smoking, and alcohol intake were not significantly associated with new-onset breast discomfort in adjusted analyses. CONCLUSIONS: Strenuous exercise and higher body weight may decrease the odds of new-onset breast discomfort among postmenopausal women initiating HT. PMID- 20009964 TI - Acellular pertussis vaccine at birth and one month induces antibody responses by two months of age. AB - BACKGROUND: Infants less than 3 months of age are at highest risk of hospitalization and death from pertussis. Several studies have examined antibody responses to pertussis vaccines at birth but no previous study has evaluated 2 doses of monovalent acellular pertussis vaccine (aPV) before 2 months of age. METHODS: Seventy-six newborns were randomized at birth to 3 groups-aPV at birth and 1 month, aPV at birth, and control. All infants received hepatitis B vaccine (HBV) at birth followed at 2, 4, and 6 months by a combination vaccine including aPV, diphtheria, tetanus, Haemophilus influenzae type b (Hib), hepatitis B, polio antigens and 7 valent conjugate pneumococcal vaccine. IgG antibody responses to pertussis toxoid (PT), filamentous hemagglutinin (FHA), and pertactin (PRN) were measured in maternal serum and in infants at 2, 4, 6, and 8 months of age. Antibody responses to hepatitis B, diphtheria, tetanus, and Hib were measured at 8 months only. A parental diary and active telephone follow-up occurred for 7 days after each vaccination. RESULTS: The aPV birth dose was well tolerated. By 2 months of age, 22 of 25 (88%) of 2 dose recipients had detectable IgG antibody to PT (IgG PT) compared with 9 of 21 (43%) who received a birth dose only and 3 of 20 (15%) of controls. Infants in the 2 dose group had a geometric mean concentration (GMC) of IgG PT of 16 ELISA units per mL (EU/mL), 95% CI: 11 to 25, significantly higher than birth dose only (5 EU/mL, 95% CI: 3-8) and controls (3 EU/mL, 95% CI: 2-5). At 8 months of age, following 5, 4, and 3 doses of aP containing vaccine, respectively, IgG PT had plateaued but IgG to FHA and PRN increased with successive doses. There was a trend to lower antibody responses for hepatitis B and Hib with higher numbers of Pa doses. CONCLUSION: These data suggest that aPV at birth and 1 month induces significantly higher IgG antibody against pertussis antigens by 2 months of age without reducing subsequent pertussis antibody responses. Larger and more detailed studies of aPV from birth are needed to evaluate other antibody responses and the potential of this approach to reduce death and morbidity from Bordetella pertussis infection in the first 3 months of life. PMID- 20009966 TI - Prevalence of antinuclear antibodies in 3 groups of healthy individuals: blood donors, hospital personnel, and relatives of patients with autoimmune diseases. AB - Antinuclear antibodies (ANA) are frequently found in healthy populations. To define the prevalence, pattern, and titer of ANA in different groups of the healthy Mexican population, we studied 304 individuals, classified into 3 groups: 104 blood donors, 100 hospital personnel working at The State General Hospital, which included doctors, laboratory technicians, and nurses; and 100 relatives of patient diagnosed either with systemic lupus erythematosus or rheumatoid arthritis, all of them apparently healthy at the time of study. We determined ANA using immunofluorescence microscopy performed on HEp-2 cells. Fluorescence was detected in 165 serum samples (54.3%). The most frequent pattern was the speckled (50.3%). The most frequent dilution was 1:40 (35.4%), followed by 1:80 (13.4%), 1:160 (3.2%), and 1:320 (1.3%).Regarding the results by study group, we found a trend toward higher ANA levels in group 2 (hospital personnel), compared with group 1 (blood donors) and group 3 (relatives of patients), a trend also reflected by the increasing frequency of serum titers of 1:80 and higher (P = 0.074). According to occupation, medical doctors showed a higher incidence of speckled pattern when compared with other occupations (P = 0.022). Medical doctors (n = 75) showed also higher titers of this particular pattern (P = 0.03). In group 3, relatives of patients with systemic lupus erythematosus showed the speckled pattern more frequently than relatives of patients with rheumatoid arthritis, in low titers (P = 0.017). We suggest that ANA tests showing speckled pattern should be at a 1:160 titer or higher to be considered positive; other patterns such as homogeneous, peripheral, or centromeric might be considered positive even at low titers ( .90). Convergent validity of the teamwork tool was measured by correlating the Teamwork subscale of the Safety Attitudes Questionnaire with the NTS (r = .76, p < .01). The NTS had good test-retest reliability (r = .92 for overall 33 items; r = .77 to.87 for the five subscales) and internal consistency (alpha = .94 for overall items; alpha = .74 to.85 for the subscales). Aggregation of individual-level responses to the unit level was supported by intraclass correlation coefficient 1 = .16 (p < .001), intraclass correlation coefficient 2 = .9 (p < .001), and mean rWG(J) = .98. DISCUSSION: The NTS was demonstrated to have good psychometric properties. Further NTS research should include testing the tool in hospitals with varying characteristics and exploring the links to clinical and operational outcomes. PMID- 20010045 TI - Research coordinators' experiences with scientific misconduct and research integrity. AB - BACKGROUND: Most reports of scientific misconduct have been focused on principal investigators and other scientists (e.g., biostatisticians) involved in the research enterprise. However, by virtue of their position, research coordinators are often closest to the research field where much of misconduct occurs. OBJECTIVE: The purpose of this study was to describe research coordinators' experiences with scientific misconduct in their clinical environment. DESIGN: The descriptive design was embedded in a larger cross-sectional national survey. A total of 266 respondents, predominately registered nurses, who answered "yes" to having firsthand knowledge of scientific misconduct in the past year, provided open-ended question responses. METHODS: Content analysis was conducted by the research team, ensuring agreement of core categories and subcategories of misconduct. FINDINGS: Research coordinators most commonly learned about misconduct via firsthand witness of the event, with the principal investigator being the person most commonly identified as the responsible party. Five major categories of misconduct were identified: protocol violations, consent violations, fabrication, falsification, and financial conflict of interest. In 70% of cases, the misconduct was reported. In most instances where misconduct was reported, some action was taken. However, in approximately 14% of cases, no action or investigation ensued; in 6.5% of cases, the coordinator was fired or he or she resigned. CONCLUSIONS: This study demonstrates the need to expand definitions of scientific misconduct beyond fabrication, falsification, and plagiarism to include other practices. The importance of the ethical climate in the institution in ensuring a safe environment to report and an environment where evidence is reviewed cannot be overlooked. PMID- 20010046 TI - Complementary and alternative medicine use, spending, and quality of life in early stage breast cancer. AB - BACKGROUND: Up to 80% of women with breast cancer use complementary and alternative medicine (CAM) therapies to improve quality of life (QOL) during treatment. OBJECTIVE: The objective of this study was to explore the association between CAM therapy use, spending on CAM therapies, demographic variables, surgical treatment, and QOL. METHODS: A secondary analysis was conducted for women with early stage breast cancer (N = 222) who were enrolled in a nursing intervention study after breast surgery. The complementary therapy utilization instrument included specific CAM therapies used, number of treatments, and spending. The number of major categories of CAM and the specific CAM therapies used were analyzed in relation to study variables using proportional odds logistic regression models. The outcome of spending for major CAM categories was analyzed using linear mixed effects modeling. RESULTS: A majority of women (56.8%) used at least one CAM therapy, and the biologically based therapies category was most frequently used (43.7%). Women with less than a college level of education were less likely to use CAM (odds ratio [OR] = 0.36, 95% confidence interval [CI] = 0.15-0.86, p < .01). Women who were employed were more likely to use therapies from multiple CAM categories (OR = 2.42, 95% CI = 1.00-5.88, p < .05), and those with lower QOL were more likely to use CAM (OR = 0.97, 95% CI = 0.95-0.99, p < .01). The results support that women with early stage breast cancer, especially those with lower QOL, are highly likely to use CAM therapies. Further research is needed on CAM interventions to enhance supportive care for breast cancer. PMID- 20010047 TI - Perceived influences on physical activity and diet in low-income adults from two rural counties. AB - BACKGROUND: Despite the increased use of ecological models in health behavior research, multilevel influences on health behaviors in rural, low-income people, an aggregate at high risk for sedentary behavior and inadequate diets, have been examined in few studies. OBJECTIVE: The purpose of this study was to describe influences on physical activity and diet in low-income, rural adults. METHOD: A cross-sectional survey was conducted using face-to-face interviews in a convenience sample of 137 low-income Anglo and Latino adults recruited from two rural Wisconsin counties. The survey included questions on health behaviors, self efficacy, barriers, social support, and community environments. Self-report data on physical activity and fruit and vegetable intake were categorized into outcome variables of meets recommendation or does not meet recommendation. Latent class cluster analysis was used to identify clusters of participants with similar influences on health behaviors, and cluster membership was used as an independent variable in logistic regression of physical activity and diet outcomes. RESULTS: Fifty-two percent of participants met a recommendation for physical activity, but only 8% met their MyPyramid recommendation for fruit and vegetable intake. Participants in the moderate self-efficacy/high safety cluster were significantly more likely than those in the low self-efficacy/moderate safety cluster to meet a recommendation for physical activity (odds ratio = 2.65). For healthy diet, participants in the low barriers cluster were significantly more likely to eat more fruits and vegetables (odds ratio = 4.13) than those in the high barriers cluster. DISCUSSION: People with healthier behaviors were distinguished from those with less healthy behaviors by higher levels of intrapersonal, interpersonal, and community supports. Results support the importance of multilevel approaches to promoting healthy lifestyles in rural, low-income adults. PMID- 20010059 TI - Advancements in body magnetic resonance imaging. PMID- 20010060 TI - Magnetic resonance nephrourographic techniques and applications: how we do it. AB - Chronic kidney disease is a significant public health problem, and a comprehensive evaluation of renal disease often requires accurate evaluation of both kidney structure and function. Magnetic resonance (MR) nephrourography refers to newly developed imaging techniques that have the ability to provide a quantitative assessment of renal function, especially glomerular filtration rate and renal blood flow. Our review outlines several different methodologies that are present in the literature and also details the specifics of our own methods for renal imaging. Though varied, all MR imaging methods use the common steps of image acquisition, image postprocessing, and tracer kinetics modeling of the processed image data. The optimal methodology should be practical and based primarily on simplicity, speed, and reproducibility. The combination of anatomic and quantitative functional information of the kidneys provided by MR imaging allows for a safe, comprehensive evaluation of renal disease, with particular utility in the settings of urinary tract obstruction and renal transplantation. PMID- 20010061 TI - Gadolinium-enhanced imaging of liver tumors and manifestations of hepatitis: pharmacodynamic and technical considerations. AB - The ability for contrast-enhanced magnetic resonance imaging to provide significant diagnostic impact to focal and diffuse liver diseases requires knowledge, analysis, and technical optimization of the imaging techniques. Our review outlines the technical requirements needed to perform reproducible contrast-enhanced liver imaging and describes the important imaging features for assessing liver disease with conventional and alternate gadolinium-based contrast media. We present an experimental review of timing and quantification methods in dynamic contrast-enhanced liver imaging, with results of analysis showing perfusion and uptake curves in a series of patients and healthy subjects. An evidence-based methodology for reproducible arterial-phase imaging is detailed for performing a real-time bolus-tracking method. Additional diagnostic imaging features manifest at later imaging phases, in which the kinetic behavior of the contrast media serves to further specify focal lesions, while revealing detailed information of diffuse liver disease, particularly hepatic fibrosis. We review the utility of alternate gadolinium-based contrast media that undergo hepatocyte uptake, for applications related to liver tumor imaging. We also introduce results showing the potential for using alternate hepatocyte uptake agents to detect and quantify liver changes related to acute and chronic hepatitides. PMID- 20010062 TI - Abdominal magnetic resonance elastography. AB - Magnetic resonance elastography (MRE) is a magnetic resonance imaging-based technique for quantitatively assessing the mechanical properties of tissues based on the propagation of shear waves. Multiple studies have described many potential applications of MRE, from characterizing tumors to detecting diffuse disease processes. Studies have shown that MRE can be successfully implemented to assess abdominal organs. The first clinical application of MRE to be well documented is the detection and characterization of hepatic fibrosis, which systematically increases the stiffness of liver tissue. In this diagnostic role, it offers a safer, less expensive, and potentially more accurate alternative to invasive liver biopsy. Emerging results suggest that measurements of liver and spleen stiffness may provide an indirect way to assess portal hypertension. Preliminary studies have demonstrated that it is possible to use MRE to evaluate the mechanical properties of other abdominal structures, such as the pancreas and kidneys. Steady technical progress in developing practical protocols for applying MRE in the abdomen and the pelvis provides opportunities to explore many other potential applications of this emerging technology. PMID- 20010063 TI - Review of magnetic resonance spectroscopy in the liver and the pancreas. AB - Proton magnetic resonance spectroscopy is a powerful tool for in vivo biochemical characterization of normal and abnormal tissues. The initial application in the abdomen was the measurement of fat concentration in the liver using chemical shift imaging. The success of chemical shift imaging in providing a semiquantitative measure of liver fat concentration led to the application of the more quantitative single-voxel volume-selective spectroscopy of the liver. This single-voxel volume-selective spectroscopic technique is able to characterize the different lipids and metabolites present in the liver and the pancreas, providing information about the ratio of unsaturated and saturated lipids. The purposes of this article were to review the spectroscopic techniques and to discuss some of the clinical applications of these techniques in the abdomen. PMID- 20010064 TI - Magnetic resonance measurement of diffusion in the abdomen. AB - Diffusion-weighted (DW) magnetic resonance imaging is an emerging noninvasive technique increasing its spectrum of use in the abdomen. Diffusion-weighted imaging has been used as add-on to routine abdominal protocol because it may potentially substitute contrast-enhanced imaging in cases under risk of nephrogenic systemic fibrosis. The apparent diffusion coefficient (ADC) images calculated from DW images enable qualitative and quantitative evaluations of tissue water mobility and functional environment because of changes in intracellular, extracellular, and intravascular tissue compartments. This article presents the basic physics of the ADC measurement, the techniques for performing ADC measurements of the liver and the pancreas, and the clinical applications of DW imaging. PMID- 20010065 TI - Dynamic contrast-enhanced magnetic resonance imaging in prostate cancer. AB - Prostate cancer remains a major health concern for the male population. During the past decade, a dramatic increase in prostate-specific antigen and transurethral resection of the prostate has resulted in increased detection rate of small lesions and increased incidence of this disease. Needle biopsies in asymptomatic men have also contributed to the increased incidence of prostate cancer, leading to an increasing incidence-to-mortality ratio. Magnetic resonance imaging (MRI) is the modality of choice in prostate cancer patients with increased prostate-specific antigen levels before or after prostate cancer diagnosis confirmed by biopsy and T2-weighted imaging (T2W) has been used as a standard technique in detection. During the last decade, dynamic contrast enhanced MRI has emerged as one of the main techniques used in multiparametric MRI of the prostate gland in cancer patients. Dynamic contrast-enhanced MRI has been routinely used for detection and diagnosis of the tumor, for staging and monitoring the therapeutic response, as well as for guiding targeted biopsies in patients with suspected prostate cancer or with a negative biopsy result. In this article, we are going to review the analysis techniques of dynamic contrast enhanced MRI and its various clinical applications in prostate cancer patients. PMID- 20010066 TI - Use of magnetic resonance imaging contrast agents to detect transplanted liver cells. AB - Liver transplantation saves the lives of millions of patients every year. The advent of cell rather than organ transplantation could potentially further improve the success of this approach. However, one problem facing the delivery and the monitoring of cell transplants is their noninvasive in vivo visualization. Noninvasive imaging is needed for this. To distinguish transplanted cells from the host liver, it is necessary to either tag these using exogenous contrast agents (eg, iron oxide nanoparticles) or insert a reporter gene that could selectively identify transplanted cells. Nevertheless, these approaches face significant challenges such as providing sufficient signal-to noise, cellular toxicity, or unequivocal detection. Preclinical studies are currently under way to refine these approaches with initial clinical trials being on the horizon for the next few years. A gradual refinement of these approaches and a robust clinical implementation promise a significant step in ensuring greater efficacy of cell transplants for the diseased liver. PMID- 20010067 TI - The nursing shortage as a community transformational opportunity: an update. AB - In the 2002 original work discussing some of our professional issues that have contributed to the current nursing shortage, I concluded that until the profession examines and addresses some of the troublesome and paradoxical areas of our workplaces, we will continue to remain locked in our current cyclical strange loop of nursing shortages and a dissatisfied nursing working force. While the nursing shortage seems to have taken on a new context, given current global economic issues and healthcare reformation efforts, many of the professional issues I addressed in my original article remain pertinent to our professional growth and the future of an autonomous workforce of nurses. In this article, the writer will briefly review the current literature around the nursing shortage, reexplore our own role in contributing to the cyclical or strange loop shortage of professional nurses, and further discuss possible solutions for the growth of nursing as a profession. The aim of this article is to explore how we can address our professional identity issues in ways that will serve to manage the current challenges we face of attracting and retaining a stable workforce. PMID- 20010068 TI - Othering: difference understood??: a 10-year analysis and critique of the nursing literature. AB - It has been 10 years since Advances in Nursing Science (ANS) published the first article that articulated othering within a nursing framework. This issue's topic, Critique and Replication, provided an opportunity for analysis of the influence of the original ANS article on the visibility and application of othering within nursing literature. Comprehensive searches of major health databases identified 32 publications citing the original ANS article. The analysis focused on exclusionary and inclusionary othering, role taking, and constructions of the other. Results indicate that although othering has become more visible within nursing literature, exclusionary othering dominates. The article concludes with thoughts for moving beyond othering and toward engagement. PMID- 20010069 TI - Comparison of interferon gamma and interferon gamma-inducible protein-10 secretion in HIV-tuberculosis patients. AB - Interferon gamma (IFNgamma)-based in-vitro assays have suboptimal sensitivity, especially in immunocompromised individuals, which emphasizes the need for alternative markers for tuberculosis (TB) diagnosis. We compared TB antigens specific IFNgamma and IFNgamma-inducible protein-10 levels in culture of whole blood samples from HIV-TB patients. We report that IFNgamma-inducible protein-10 detects a greater number of HIV-TB cases than IFNgamma and suggest that IFNgamma inducible protein-10 may be a better alternative marker for latent TB infection diagnosis among immunocompromised individuals. PMID- 20010070 TI - The MONET trial: darunavir/ritonavir with or without nucleoside analogues, for patients with HIV RNA below 50 copies/ml. AB - BACKGROUND: In virologically suppressed patients, darunavir-ritonavir (DRV/r) monotherapy could maintain virological suppression similarly to DRV/r and two nucleosides. METHODS: Two hundred and fifty-six patients with HIV RNA less than 50 copies/ml for over 24 weeks on current antiretrovirals [non-nucleoside reverse transcriptase inhibitor (NNRTI)-based (43%), or protease inhibitor-based (57%)], switched to DRV/r 800/100 mg once daily, either as monotherapy (n = 127) or with two nucleoside reverse transcriptase inhibitors (NRTIs) (n = 129). Treatment failure was defined as two consecutive HIV RNA levels above 50 copies/ml (TLOVR) by week 48, or switches off study treatment. The trial had 80% power to show noninferiority for the monotherapy arm (delta = -12%). RESULTS: Patients were 81% male and 91% Caucasian, with mean age 44 years, and CD4 cell count of 574 cells/microl. In the primary efficacy analysis, HIV RNA less than 50 copies/ml by week 48 (per protocol) was 86.2 versus 87.8% in the monotherapy and triple therapy arms; by intent-to-treat switch equals failure, efficacy was 84.3 versus 85.3%; by a switch-included analysis, efficacy was 93.5 versus 95.1%: all three comparisons showed noninferior efficacy for DRV/r monotherapy. CD4 cell counts remained stable during the trial in both arms. One patient per arm showed at least one protease inhibitor mutation, and one patient in the triple therapy arm showed an NRTI mutation. Nine patients per arm discontinued randomized treatment for either adverse events or other reasons. No new or unexpected safety signals were detected. CONCLUSIONS: In this study for patients with HIV RNA less than 50 copies/ml on other antiretrovirals at baseline, switching to DRV/r monotherapy showed noninferior efficacy versus triple antiretroviral therapy. PMID- 20010072 TI - Transmission networks of HIV-1 among men having sex with men in the Netherlands. AB - OBJECTIVE: To obtain insight in the HIV-1 transmission networks among men having sex with men (MSM) in the Netherlands. DESIGN: A phylogenetic tree was constructed from polymerase sequences isolated from 2877 HIV-1 subtype B-infected patients monitored as part of the AIDS Therapy Evaluation in the Netherlands (ATHENA) nationwide observational cohort. METHODS: For MSM with a known date of infection, the most similar sequences were selected as potential transmission pairs when they clustered with bootstrap value of at least 99%. Time from infection to onward transmission was estimated as the median time between dates of infection for each transmission pair. The source of infections with a resistant strain was traced using the entire phylogenetic tree. RESULTS: Of sequences from 403 MSM with a known date of infection between 1987 and 2007, 175 (43%) formed 63 clusters. Median time to onward transmission was 1.4 years (interquartile range 0.6-2.7). Twenty-four (6%) MSM carried a virus with resistance-related mutations, 13 of these were in eight clusters together with sequences from 28 other patients in the entire phylogenetic tree. Six clusters contained sequences obtained from 29 men all presenting the same resistance related mutations. CONCLUSION: From our selection of likely transmission pairs, we conclude that onward transmission of HIV-1 from infected MSM in the Netherlands happens both during and after primary infection. Transmission of resistant strains from the antiretroviral therapy-treated population is limited, but strains with resistance-related mutations have formed subepidemics. PMID- 20010075 TI - HIV delivers its infectious cargo concealed within an endosome. PMID- 20010074 TI - N348I in HIV-1 reverse transcriptase decreases susceptibility to tenofovir and etravirine in combination with other resistance mutations. AB - We previously demonstrated that N348I in HIV-1 reverse transcriptase confers zidovudine and nevirapine resistance. However, both of these inhibitors are currently infrequently used in developed countries, and the impact of N348I on newer reverse transcriptase inhibitors, such as tenofovir and etravirine, is unknown. In this study, we demonstrate that N348I alone confers no resistance to tenofovir and low-level resistance to etravirine. However, N348I significantly decreases tenofovir susceptibility when combined with thymidine analogue mutations and etravirine susceptibility when combined with Y181C. PMID- 20010076 TI - Optimal equations for describing the relationship between prostate volume, number of sources, and total activity in permanent prostate brachytherapy. AB - OBJECTIVES: To determine whether there is an optimal type of mathematical equation for predicting seed and activity requirements for permanent prostate brachytherapy. METHODS: Four institutions with extensive brachytherapy experience each submitted details of more than 40 implants. The data was used to generate power and linear equations to reflect the relationship between preimplant volume and the number of seeds implanted, and preimplant volume and the total implant activity. We compared the R and standard error of the generated equations to determine which type of equation better fit the data. RESULTS: For the limited range of prostate volumes commonly implanted (20-60 mL), power and linear equations predict seed and activity requirements comparably well. CONCLUSIONS: Linear and power equations are equally suitable for generating institution specific nomograms. PMID- 20010077 TI - A volumetric trend analysis of the prostate and seminal vesicles during a course of intensity-modulated radiation therapy. AB - PURPOSE: The purpose of this study was to evaluate the volumetric changes of the prostate and seminal vesicles (SV) during a definitive course of intensity modulated radiation therapy using a combined computed tomography (CT)-linear accelerator system. METHODS AND MATERIALS: Fifteen patients were enrolled in this IRB-approved prospective study. The treatment plan was designed to deliver a total dose of 75.6 Gy over 42 fractions to 98% of the planning target volume. In room CT scans were acquired using a CT-on-rails system 3 times weekly just before daily treatment throughout the course of radiation therapy. Rates of volume change over time for the prostate and SV were calculated using linear regression analysis. RESULTS: Fifteen patients had a total of 369 CT scans during the study. For 53% of patients (N = 8), there was a statistically significant decrease in prostate volume over time (range, 0.05-0.47 mL/d or 0.7%-3.5% per week). The median volume reduction was -11.5% (4.9 mL) at the end of the treatment. There was no significant change in the volume of the SV during the course of treatment. The decrease in prostate volume was significantly correlated with the initial volume of the prostate, with larger glands shrinking more during treatment (P = 0.001). CONCLUSION: Patients with enlarged prostates may experience volumetric reduction in their gland size during a protracted course of radiation therapy. However, the magnitude of the volume reduction is relatively small. In the current era of dose escalation, future studies with adaptive radiation therapy strategy may be appropriate to minimize radiation exposure to normal tissues. PMID- 20010071 TI - Targeting Trojan Horse leukocytes for HIV prevention. PMID- 20010078 TI - Is radical prostatectomy the "gold standard" for localized prostate cancer? AB - The purpose of this article is to review the pertinent literature and compare the outcomes after radical prostatectomy (RP) and radiotherapy (RT). The probability of cure is similar after either RP or RT. The likelihood of significant treatment complications is probably higher after RP compared with RT. Preservation of erectile function is at least as good or better with RT compared with RP. Urinary continence is more likely to be preserved after RT compared with RP. Each treatment results in distinct patterns of adverse changes in quality of life that are worsened by factors including obesity, large prostate size, high prostate specific antigen (PSA), and older age. PMID- 20010073 TI - Response to planned treatment interruptions in HIV infection varies across childhood. AB - OBJECTIVE: To evaluate clinical, immunological and virological consequences of CD4-guided antiretroviral therapy (ART) planned treatment interruptions (PTIs) compared with continuous therapy in children with chronic HIV infection in the Paediatric European Network for Treatment of AIDS 11 trial. DESIGN: This was a multicentre, 72-week, open, randomized, phase II trial. METHODS: One hundred and nine children with HIV-RNA below 50 copies/ml and CD4% of at least 30% (2-6 years) or at least 25% and CD4 cell count of at least 500 cells/microl (7-15 years) were randomized to continuous therapy (53) or PTI (56). In PTI, ART was restarted if confirmed CD4% was less than 20% or more than 48 weeks had been spent off ART. The primary outcome was Centers for Disease Control and Prevention (CDC) stage C event, death or CD4% less than 15% (and CD4 cell count less than 200 cells/microl for children aged 7-15 years). RESULTS: At baseline, median (interquartile range) age was 9 (6-12) years, CD4% 37% (33-41), CD4 cell count 966 (793-1258) cells/microl, nadir CD4% before combination ART 18% (10-27), time on ART 6 (3-6) years and 26% were CDC stage C. After median (range) 130 (33-180) weeks of follow-up, 4 versus 48% of time was spent off ART in continuous therapy and PTI, respectively. No child died or had a new CDC stage C event; one (2%) continuous therapy versus four (7%) PTI children had a primary outcome based on CD4%/cell count (P = 0.2). Lower nadir CD4% predicted faster CD4% decline after stopping ART. Younger age and higher nadir CD4% predicted being off ART for at least 48 weeks and better CD4% recovery following PTI. CONCLUSION: In this first paediatric trial of PTI, there were no serious clinical outcomes. Younger children had better CD4% recovery after PTIs. Immunology substudies and long-term follow-up in Paediatric European Network for Treatment of AIDS 11 trial are ongoing. Further research into the role of treatment interruption in children is required, particularly, as guidelines now recommend early ART for all infected infants. PMID- 20010079 TI - Patient recruitment methods to evaluate treatment decision making for localized prostate cancer. AB - OBJECTIVE: To examine methods for identifying and recruiting prostate cancer patients prior to initiating treatment to gain insight into the treatment decision process and avoid recall bias. STUDY DESIGN AND SETTING: One recruitment strategy involved providers and nursing staff approaching patients in community and academic urology clinics. The second recruitment strategy used electronic pathology reports to identify newly diagnosed cases in real time in an integrated health system. Our recruitment goal was to have patients complete the survey about the decision-making process prior to initiating therapy. RESULTS: The two recruitment methods yielded different response rates. Of the 226 eligible participants approached in urology clinics, 187 (83%) returned a completed baseline survey. Of the 1177 surveys mailed to potentially eligible participants at KPNC, 617 (52%) returned a completed baseline survey. The number of surveys completed prior to treatment was 125 (67%) for the clinic recruitment approach and 437 (71%) for the electronic medical record approach. Younger participants and patients with less aggressive clinical characteristics were more likely to complete the survey before initiating treatment. Other patient demographic and clinical factors were not associated with the timing of survey return. CONCLUSIONS: Recruiting newly diagnosed patients prior to initiating treatment is feasible with both approach methods. The use of electronic medical records to identify subjects was more cost efficient, although it results in a lower response rate. PMID- 20010082 TI - Prevention of parastomal hernia. Is it possible? PMID- 20010080 TI - Genealogical tree study as screening method in the Lynch syndrome prior to genetic test. AB - OBJECTIVES: Despite genetic advances in the study of Lynch syndrome (LS), difficulties remain in the diagnosis of the syndrome. The aim of this study was to assess the usefulness of a detailed genealogical tree as a screening method to identify Tenerife island families with a high probability of LS. METHODS: We elaborated complete genealogical trees of the families. According to the degree of fulfillment of the Amsterdam Criteria II, the genealogical trees were classified as high or low probability of LS. Additionally, we analyzed the level of tumor microsatellite instability (MSI+) and identified a mutation in exon 13 of the MSH2 gene by single-strand conformation polymorphism, sequencing, and PCR RFLP. RESULTS: According the genealogical trees, we found 10 families with high probability of LS and 30 families with low probability of LS. The families with high probability of LS showed high MSI+ in all cases. Conversely, families with low probability were MSS (microsatellite stable). In 5 of the 10 families with high probability, we discovered a T-->G mutation in position 688 of exon 13 of MSH2, which appeared in all the family members with the tumor, except 1 patient with a retinoblastoma. CONCLUSIONS: Our results indicate that genealogical tree is a highly effective tool for classifying families with a high probability of Lynch Syndrome prior to genetic test. PMID- 20010083 TI - Prognostic value of circulating mutant DNA in unresectable metastatic colorectal cancer. AB - OBJECTIVE: No validated biologic prognostic marker is presently available in metastatic colorectal cancer (MCRC). We prospectively evaluated the prognostic value of circulating mutant DNA in 31 patients presenting an unresectable MCRC treated by chemotherapy, and we used, as tumor markers, KRAS mutations and methylation of the RASSF2A promoter. METHODS: Detection in the serum of KRAS mutation and RASSF2A methylation were performed using sensitive methods, respectively, real-time polymerase chain reaction (PCR) performed in the presence of a peptide nucleic acid specific of the wild-type sequence and methyl-specific PCR after bisulfite treatment. RESULTS: Among 29 MCRC patients for whom DNA from the primary tumor was available, 23 (79%) presented at least one of the markers in their primary tumor, and 12 of them presented the same alteration in serum. For the 2 remaining patients, RASSF2A methylation was detected in serum indicating that this alteration was present in the primary tumor. These 14 patients with a detectable tumor marker in their serum were designed sDNA+ patients. After 6 months of follow-up, 11/14 (79%) sDNA+ and 1/11 (9%) sDNA- patients presented a progressive disease (P = 0.001). The median progression free survival was 5 months in sDNA+ patients versus 14 months in sDNA- patients (P = 0.004). After 1 year of follow-up, 2 of 14 (14%) sDNA+ and 8 of 11 (73%) sDNA- patients presented no signs of disease progression (P = 0.005). CONCLUSIONS: This study suggests that the presence of circulating mutant DNA in unresectable MCRC patients, which can be detected using simple methods such as methylation-specific PCR or real-time PCR, is highly predictive of clinical outcome. PMID- 20010084 TI - A comprehensive review of topical hemostatic agents: efficacy and recommendations for use. AB - Since ancient times we have attempted to facilitate hemostasis by application of topical agents. In the last decade, the number of different effective hemostatic agents has increased drastically. In order for the modern surgeon to successfully choose the right agent at the right time, it is essential to understand the mechanism of action, efficacy and possible adverse events as they relate to each agent. In this article we provide a comprehensive review of the most commonly used hemostatic agents, subcategorized as physical agents, absorbable agents, biologic agents, and synthetic agents. We also evaluate novel hemostatic dressings and their application in the current era. Furthermore, wholesale acquisition prices for hospitals in the United States are provided to aid in cost analysis. We conclude with an expert opinion on which agent to use under different scenarios. PMID- 20010085 TI - Chimeric composite skin substitutes for delivery of autologous keratinocytes to promote tissue regeneration. AB - OBJECTIVE: We hypothesize that the pathogen-free NIKS human keratinocyte progenitor cell line cultured in a chimeric fashion with patient's primary keratinocytes would produce a fully stratified engineered skin substitute tissue and serve to deliver autologous keratinocytes to a cutaneous wound. SUMMARY OF BACKGROUND DATA: Chimeric autologous/allogeneic bioengineered skin substitutes offer an innovative regenerative medicine approach for providing wound coverage and restoring cutaneous barrier function while delivering autologous keratinocytes to the wound site. NIKS keratinocytes are an attractive allogeneic cell source for this application. METHODS: Mixed populations of green fluorescent protein (GFP)-labeled NIKS and unlabeled primary keratinocytes were used to model the allogeneic and autologous components in chimeric monolayer and organotypic cultures. RESULTS: In monolayer coculture, GFP-labeled NIKS had no effect on the growth rate of primary keratinocytes and cell-cell junction formation between labeled and unlabeled keratinocytes was observed. In organotypic culture employing dermal and epidermal compartments, chimeric composite skin substitutes generated using up to 90% GFP-labeled NIKS exhibited normal tissue architecture and possessed substantial regions attributable to the primary keratinocytes. Tissues expressed proteins essential for the structure and function of a contiguous, fully-stratified squamous epithelia and exhibited barrier function similar to that of native skin. Furthermore, chimeric human skin substitutes stably engrafted in an in vivo mouse model, with long-term retention of primary keratinocytes but loss of the GFP-labeled NIKS population by 28 days after surgical application. CONCLUSIONS: This study provides proof of concept for the use of NIKS keratinocytes as an allogeneic cell source for the formation of bioengineered chimeric skin substitute tissues, providing immediate formal wound coverage while simultaneously supplying autologous cells for tissue regeneration. PMID- 20010086 TI - Regional trauma system reduces mortality and changes admission rates: a before and after study. AB - OBJECTIVE: To evaluate the effect of the introduction of a regionalized trauma system. BACKGROUND: Trauma systems have proven to be efficacious in reducing mortality in trauma patients in the United States. To date, this was not proven for inclusive trauma systems outside the United States. The current study evaluates the effect of the introduction of an inclusive trauma system in the Netherlands in 1999. METHODS: Retrospective pre- and post analyses of a trauma care system on hospital discharge data regarding trauma patients admitted to hospitals in the central region of The Netherlands. Patients treated during 1996 to 1998 (control group), before implementation of the inclusive trauma system were compared with patients treated during 2003 to 2005 (index group) after the trauma system was installed. Risk adjusted odds-ratios of death and admission to a trauma center were determined. RESULTS: A total of 33,201 patients were included in the control group and compared with 34,840 patients in the index group. After implementation of the trauma system, in-hospital mortality for all injured patients decreased from 2.6% to 2.3% (OR: 0.89 with 95% CI: 0.80-0.98). After adjustment for differences in gender, age, injury severity, comorbidity, injured body region, mechanism and intent of injury between both groups, the odds ratio was 0.84 with 95% CI (0.76-0.94). Multitrauma patients were the subgroup admitted more frequently to a trauma center (OR: 1.19 with 95% CI: 1.01-1.39). CONCLUSIONS: Implementation of an inclusive trauma system in The Netherlands results in a more efficient triage system of trauma patients among hospitals and is associated with a substantial and statistically significant risk reduction (16%) of death. PMID- 20010088 TI - The impact of gastric bypass on gastroesophageal reflux disease in patients with morbid obesity: a prospective study based on the Montreal Consensus. AB - OBJECTIVES: To assess the impact of gastric bypass (GBP) on gastroesophageal reflux disease (GERD) based on Montreal Consensus. METHODS: In this study, 86 patients (25 men; aging 38 +/- 12 years; body mass index 45 [35-68 kg/m2]) were investigated for GERD before GBP and 6 months later. Esophageal and extraesophageal syndromes were assessed based on Montreal Consensus. Esophageal acid exposure and gastric pouch acidity were also evaluated. RESULTS: Overall prevalence of GERD was 64% before GBP and 33% after GBP (P < 0.0001). Typical reflux syndrome (TRS) was present in 47 patients (55%) preoperatively and disappeared in 39 of them (79%) post-GBP. Out of 39 patients with no symptoms, 4 (10%) developed TRS postoperatively (P < 0.0001). The chief TRS complaint changed from heartburn pre-GBP (96%) to regurgitation post-GBP (64%). Esophageal mucosa improved in 27, was unchanged in 51, and worsened in 8 patients (P = 0.001) in regard of esophagitis. Extraesophageal syndromes were present in 16 patients preoperatively and in none but one post-GBP (P = 0.0003). GERD-related well being and use of proton pump inhibitors were both improved after GBP. Total acid exposure decreased from a median (interquartile range, 25%-75%) of 5.1% (range, 2 8.2) to 1.1% (range, 0.2-4.8), P = 0.0002. Most patients (86%) showed and acid gastric pouch in fasting conditions post-GBP. CONCLUSIONS: GBP ameliorated GERD syndromes in most patients 6 months after the procedure, resulting in quality of life improvement and less proton pump inhibitors usage. Whether regurgitation post-GBP corresponds to reflux disease or bad eating behavior deserves further studies. PMID- 20010087 TI - Putative predictive parameters for the outcome of laparoscopic splenectomy: a multicenter analysis performed on the Italian Registry of Laparoscopic Surgery of the Spleen. AB - OBJECTIVE: To identify predictive risk factors for conversion to open splenectomy and postoperative complications in patients undergoing elective laparoscopic splenectomy. BACKGROUND: The laparoscopic approach represents the "gold standard" for splenectomy, but its use in the treatment of splenomegaly and malignant disease is controversial. Factors that influence immediate outcome are clinical, anatomic, and pathologic. METHODS: Univariate and multivariate analyses of data from the Italian Registry of Laparoscopic Surgery of the Spleen, a multicenter database supported by 25 referral centers. Analysis of data (1993-2007) was performed on a series of patients (n = 676) undergoing elective laparoscopic splenectomy. Demographic data, the operative indications, the surgical technique applied, and any intra- and/or postoperative complications with respect to the patients were assessed. Records were analyzed retrospectively using the Student t test, the chi test, and logistic regression. RESULTS: Conversion to open splenectomy was necessary in 39 cases (5.8%). Perioperative deaths occurred in 3 cases (0.4%). There were no complications in 560 patients (82.8%), with a mean hospital stay of 5 days (range, 2-54). Overall, morbidity occurred in 116 patients (17.2%). Multivariate analysis found that the body mass index (P = 0.01) and the presence of hematologic malignancy (P < 0.001) were independent predictors for intraoperative complications and surgical conversion. Spleen longitudinal diameter (P = 0.001) and surgical conversion (P = 0.001) were independent predictors for the occurrence of postoperative complications. CONCLUSIONS: This large multicenter study provides evidence for the significance of predictive risk factors for intra- and postoperative complications in laparoscopic splenic surgery. Besides splenic dimensions, other factors like the patient's habitus and the specific underlying hematologic pathology should be recognized by the surgeon to reduce complications and initiate adequate treatment. PMID- 20010089 TI - Hepatic metastases from neuroendocrine tumors with a "thin slice" pathological examination: they are many more than you think... AB - OBJECTIVES: To prospectively compare the results of the pathologic examination including thin 3- to 4-mm thick serial slices of the surgical specimen in a well defined anatomic part of the liver with those of 4 different liver imaging techniques. BACKGROUNDS SUMMARY DATA: Liver metastases (LM) of neuroendocrine origin are frequent and accuracy of pretherapeutic liver imaging in detecting small-sized LM is questioned, but no study has compared the different techniques used to image large liver specimens and meticulous pathologic examination. PATIENTS AND METHODS: Eleven patients with a metastatic endocrine tumor were selected before undergoing a right hepatectomy or a left hepatectomy passing along the median hepatic vein. Before the hepatectomy, patients underwent somatostatin receptor scintigraphy, computed tomography, magnetic resonance imaging, and ultrasonography (US). An intraoperative assessment (palpation and US) was also performed. Liver specimens were systematically cut into thin serial slices, 3- to 4-mm thick. A final histologic count of the total number of LM was obtained and compared with findings from each imaging technique. RESULTS: Compared with the final histologic count of LM, fewer than 50% of the LM were detected preoperatively. The accuracy was 24% for somatostatin receptor scintigraphy, 38% for computed tomography and US, and 49% for magnetic resonance imaging (the only imaging technique that detected half the number of LM). The size of the smallest LM was not greater than 2 mm in 54% of the patients. CONCLUSION: Half the number of LM from neuroendocrine tumors are undetectable on preoperative imaging. PMID- 20010090 TI - Activating KRAS mutations and overexpression of epidermal growth factor receptor as independent predictors in metastatic colorectal cancer patients treated with cetuximab. AB - OBJECTIVE: Cetuximab, a monoclonal antibody targeting epidermal growth factor receptor (EGFR), has been proven to be efficient in metastatic colorectal cancer (mCRC); however, the therapeutic response is variable and markers predictive of response are urgently required. This study was conducted to determinate the predictive values of KRAS mutation status and EGFR expression in mCRC patients treated with cetuximab plus chemotherapy. SUMMARY BACKGROUND DATA: Clinical benefit with EGFR-targeting antibodies seems to be restricted to a particular subgroup of mCRC patients. Therefore, the identification of reliable predictive factors for mCRC patients is imperative before the introduction of targeted chemotherapy. METHODS: Ninety-five mCRC patients receiving cetuximab plus the FOLFIRI or FOLFOX-4 chemotherapy were enrolled into the present study. KRAS mutation status/EGFR expression levels were analyzed using direct sequencing, immunohistochemistry (IHC), and reverse transcription-polymerase chain reaction (RT-PCR) assay, respectively. The association between clinical response, progression-free survival (PFS) and overall survival (OS) as well as KRAS mutation status/EGFR expression levels were evaluated. RESULTS: Of 95 mCRC patients, KRAS mutations were identified in 41 cases, and EGFR overexpression (protein or mRNA levels) were observed in 78 patients. Among 41 tumors with KRAS mutation, 33 were found to be activating mutants at codons 12, 13, 15 or 18, while 8 were nonactivating mutants at codons 20, 30, or 31. Fifty-five patients responded to cetuximab plus chemotherapy, 49 were EGFR overexpression and 46 were wild-type KRAS tumor status. Patients with tumors that express high EGFR levels or harbor wild-type KRAS are more likely to have a better PFS and OS when treated with cetuximab plus chemotherapy (all P < 0.05). Furthermore, patients with nonactivating KRAS mutants in tumors had a significantly better PFS and OS than patients with activating KRAS mutants (both P < 0.05). However, for patients with wild-type KRAS tumor status, EGFR expression remains a relevant predictor of clinical response. CONCLUSIONS: The study suggests that activating KRAS mutants is a particularly important independent predictive marker in mCRC patients treated with cetuximab plus chemotherapy, of which combing activating KRAS mutants and EGFR could help to identify the subgroup of patients who are most likely to respond to cetuximab plus chemotherapy. PMID- 20010091 TI - Cooperative antithrombotic effect from the simultaneous inhibition of thrombin and factor Xa. AB - Whereas heparin functions as an antithrombotic agent by promoting antithrombin III-based inhibition of thrombin and factor Xa, there is less appreciation for the combination behavior with small-molecule, direct inhibitors of these proteases. We conducted a study in a high-shear arterial environment to explore the potential for a cooperative antithrombotic effect with a thrombin inhibitor (argatroban), a factor Xa inhibitor (YM-60828), and a dual thrombin/factor Xa inhibitor (RWJ-445167). We employed a platelet-dependent vascular injury model in which rats were subjected to an acute electrical injury to the carotid artery. Antithrombotic efficacy was measured for thrombin inhibitor argatroban and factor Xa inhibitor YM-60828 administered alone or in combination. The results indicate that there is a cooperative antithrombotic effect in vivo when both thrombin and factor Xa are inhibited simultaneously. The dual thrombin/factor Xa inhibitor RWJ 445167 was found to have potent antithrombotic activity in this high-shear environment. A comparison of results for RWJ-445167 and argatroban showed additional efficacy with RWJ-445167, suggestive of drug synergy. PMID- 20010092 TI - Evaluation of coagulation stages of hemorrhaged swine: comparison of thromboelastography and rotational elastometry. AB - Thromboelastography (TEG) or rotational thromboelastometry (ROTEM) assesses blood viscoelastic properties and clotting kinetics that can be measured by Haemoscope TEG and Pentapharm ROTEM devices using slightly different methodologies. These devices were compared by measuring blood samples associated with various degrees of coagulopathy. Blood samples, collected from swine undergoing three types of severe injury and resuscitation protocol resulting in normal, hypercoagulopathy, and hypocoagulopathy, were assessed with TEG or ROTEM before the surgical procedures, and after injury, fluid resuscitation, and simulated hospital phase. Standard clotting parameters were compared by Student's t-test at a significance of a P value less than 0.05. Regression analysis indicated a positive correlation between TEG and ROTEM for reaction time (R), clotting rate (K), and maximum amplitude (Ma) parameters. With samples of normal coagulation, R (440 +/- 136 vs. 391 +/- 73 s), K (99 +/- 39 vs. 81 +/- 20 s), and Ma (74 +/- 4 vs. 69 +/- 5 mm) were higher, whereas (alpha) (68 +/- 8 vs. 75 +/- 3 mm) was lower with TEG than ROTEM, respectively; a P value is less than 0.05. The magnitude of changes from baseline in hypercoagulable or hypocoagulable samples due to level of injury was equivalent with TEG and ROTEM indicating comparable use of the instruments. However, when samples were extremely hypocoagulopathic due to resuscitation fluid, the TEG values could not be readily determined. Overall, TEG readings were higher than ROTEM readings; this disparity between the two instruments was attenuated with hypercoaguable samples. Both devices yielded similar information regarding the status of coagulation related to trauma. Because of operating characteristics, the same instrument should be used for monitoring the same patient or study. PMID- 20010093 TI - The exacerbation component of impairment and risk in pediatric asthma. AB - PURPOSE OF REVIEW: The new guidelines for assessment of asthma control emphasize two domains: impairment and risk. Exacerbations of asthma are an important component of risk but have not received as much attention as the day-to-day symptoms that make up impairment. The purpose of this review is to report what has recently been learned about exacerbations. RECENT FINDINGS: Exacerbations occur in the context of both controlled and uncontrolled asthma. Exacerbations are a strong independent risk factor of further exacerbations. This suggests that unrecognized factors may be involved in susceptibility to exacerbations in addition to more commonly recognized triggers such as viruses, allergens, and poorly controlled asthma. Such factors may be the result of genetic variation. Recent evidence now shows a residual effect on lung function from an exacerbation event. There are no current specific intervention measures to prevent exacerbations but attention to management practices continues to look at the role of the emergency department in improving care. SUMMARY: The presence of exacerbations is considered a risk factor of the likelihood of further exacerbations and requires assessment of the triggers identified with history of exacerbations, host factors, atopic disposition, exposure to allergens, adherence to prescribed medications, and preventive measures to reduce future risk of such an event. PMID- 20010094 TI - Local allergic rhinitis: a new entity, characterization and further studies. AB - PURPOSE OF REVIEW: The purpose of this review is to describe new insights into the pathophysiology and clinical characterization of 'local allergic rhinitis', a new phenotype of rhinitis that may affect individuals previously diagnosed with nonallergic rhinitis. RECENT FINDINGS: Several studies have shown the existence of local allergic rhinitis with nasal production of specific IgE (sIgE) antibodies in the absence of atopy in over 40% of nonallergic rhinitis patients. Evidence for this entity is supported by the clinical symptoms, the local production of sIgE and a leukocyte-lymphocyte inflammatory pattern, with an increase in the nasal fluids of eosinophils, mast cells and T lymphocytes during natural exposure to aeroallergens, as well as a positive immediate and dual response to a nasal allergen provocation test with local production of tryptase and eosinophil cationic protein and an increase of nasal sIgE to inhalant allergens. SUMMARY: On the basis of these new findings, an advanced diagnostic approach is proposed in patients with symptoms suggestive of allergic rhinitis but negative results in skin prick test and serum sIgE. Detection of local sIgE in nasal secretions during natural exposure to aeorallergens and a positive nasal allergen provocation test with local production of tryptase, eosinophil cationic protein and sIgE are useful to detect patients with local allergic rhinitis. PMID- 20010095 TI - Creating and curing fatty hearts. AB - PURPOSE OF REVIEW: Diseases associated with ectopic disposition of lipids are becoming an increasingly important medical problem as the incidence of type 2 diabetes and obesity increases. One of the organs affected by lipotoxicity is the heart and this review presents an update on human and animal studies of this problem. RECENT FINDINGS: Human studies have clearly correlated heart dysfunction with the content of triglyceride. More recently human heart samples have been used to assess gene changes associated with altered lipid accumulation. Genetically altered mice have been created that develop lipotoxic cardiomyopathies and newer investigations are attempting to delineate curative therapies. SUMMARY: Human studies will confirm the metabolic changes associated with lipotoxic cardiomyopathy and, hopefully, animal studies will guide treatment options. PMID- 20010096 TI - The nutritional and clinical significance of lipid rafts. AB - PURPOSE OF REVIEW: Lipid rafts are potentially modifiable by diet, particularly (but not exclusively) by dietary fatty acids. This review examines the potential for dietary modification of raft structure and function in the immune system, brain and retinal tissue, the gut, and in cancer cells. RECENT FINDINGS: In-vitro and ex-vivo studies suggest that dietary n-3 polyunsaturated fatty acids (PUFAs) may exert immunosuppressive and anticancer effects through changes in lipid raft organization. In addition, gangliosides and cholesterol may modulate lipid raft organization in a number of tissues, and recent work has highlighted sphingolipids in membrane microdomains as potential targets for inhibition of tumor growth. The roles of fatty acids and gangliosides, especially in relation to lipid rafts, in cognitive development, age-related cognitive decline, psychiatric disorders, and Alzheimer's disease are poorly understood and require further investigation. The roles of lipid rafts in cancer, in microbial pathogenesis, and in insulin resistance are starting to emerge, and indicate compelling evidence for the growing importance of membrane microdomains in health and disease. SUMMARY: In-vitro and animal studies show that n-3 PUFAs, cholesterol, and gangliosides modulate the structure and composition of lipid rafts, potentially influencing a wide range of biological processes, including immune function, neuronal signaling, cancer cell growth, entry of pathogens through the gut barrier, and insulin resistance in metabolic disorders. The physiological, clinical, and nutritional relevance of these observations remains to be determined. PMID- 20010097 TI - Which factors influence glycemic control in the intensive care unit? AB - PURPOSE OF REVIEW: Intensive insulin therapy titrated to restore and maintain blood glucose between 80 and 110 mg/dl (4.4-6.1 mmol/l) was found to improve survival of critically ill patients in one pioneering proof-of-concept study performed in a surgical intensive care unit. The external validity of these findings was investigated. RECENT FINDINGS: Six independent prospective randomized controlled trials, involving 9877 patients in total, were unable to confirm the survival benefit reported in the pioneering trial. Several hypotheses were proposed to explain this discrepancy, including the case-mix, the features of the usual care, the quality of glucose control and the risks associated with hypoglycemia. SUMMARY: Before a better understanding and delineation of the conditions associated with and improved outcome by tight glycemic control, the choice of an intermediate glycemic target appears as a safe and effective solution. PMID- 20010098 TI - Critique of normoglycemia in intensive care evaluation: survival using glucose algorithm regulation (NICE-SUGAR)--a review of recent literature. AB - PURPOSE OF REVIEW: The publication of the long awaited results of the Normoglycaemia in Intensive Care Evaluation - Survival Using Glucose Algorithm Regulation (NICE-SUGAR) trial generated intense controversy in the area of glycemic control in the critically ill. NICE-SUGAR reported results in direct contrast to the original Leuven study and challenged the legitimacy of a mortality benefit of tight glycemic control in the intensive care unit (ICU). This review of the recent literature critically examines the salient differences between NICE-SUGAR and the original Leuven study. RECENT FINDINGS: Differences in glycemic targets within the control and intervention groups, variability with patients reaching these set targets, and the disparity in study execution and nutritional strategies are some of the methodological differences explaining the observed differences in mortality and morbidity between the two studies. The Leuven study should be viewed as a 'proof-of-concept' study with future studies aimed at confirming its finding and optimizing clinical algorithms to safely implement it in various 'real world' settings. Discrepancies in implementation and nutrition make direct comparison of NICE-SUGAR and the original Leuven study impracticable. SUMMARY: Accurate replication of the original Leuven methodology may be the limiting factor for achieving the benefits gained by intensive insulin therapy (IIT). Determination of ICU capability (physicians, nurses, standardization of equipment, etc.) is crucial to implementing tight glycemic targets. If IIT is not achievable due to adverse outcomes such as hypoglycemia, more lax and reachable glucose control should be sought. PMID- 20010099 TI - Implications of diet on nonalcoholic fatty liver disease. AB - PURPOSE OF REVIEW: This review examines the effects of diet on nonalcoholic fatty liver disease (NAFLD). This includes the effects of calories, both in excess and restricted, as well as macronutrients. RECENT FINDINGS: Recent findings suggest that short-term hypercaloric feeding leads to increased intrahepatic triglyceride (IHTG), whereas short-term hypocaloric feeding leads to decreased IHTG, despite little change in total body weight, suggesting that ongoing excess caloric delivery directly contributes to the development of NAFLD. Weight loss with either low-fat or low-carbohydrate diets can improve IHTG; however, specific macronutrients, such as fructose, trans-fatty acids, and saturated fat, may contribute to increased IHTG independent of total calorie intake. n-3 polyunsaturated fatty acids and monounsaturated fatty acids may play a protective role in NAFLD. The mechanisms behind these effects are not fully understood. SUMMARY: Diet plays a role in the pathophysiology of NAFLD. It is reasonable to advise patients with NAFLD to reduce calorie intake with either low-fat or low carbohydrate diets as well as limit intakes of fructose, trans-fatty acids, and saturated fat. PMID- 20010100 TI - Ion transport in the small intestine. AB - PURPOSE OF REVIEW: The 2009 review on small intestinal ion transport, in this series, focused on recent advances in duodenal bicarbonate secretion, the importance of scaffolding proteins and the pathophysiology of inflammation associated diarrhea. The current review focuses on advances in ion-coupled solute transport, the dynamic role of the paracellular pathway in transepithelial-fluid transport and of elucidating the cellular basis of diarrheas associated with enteric infections. RECENT FINDINGS: In understanding the cellular pathophysiology underlying diarrheal diseases, there is increased focus on the role of altering Na absorptive mechanisms as well as the role of the paracellular pathway. This is not to minimize the role of Cl-secretory pathways, especially cystic fibrosis transmembrane conductance regulator (CFTR), which continues to have pleiotropic roles in modulating other transporters. The Na-glucose cotransporter (SGLT) was the first transporter ever to be cloned. Twenty-one years later, with another first, the crystal structure of the related Na galactose transporter has been described and opens new avenues to understand structure-function relationships and intelligent drug design for transporters. SUMMARY: Progress continues to be made on integrating information obtained from reductionist models into more complex in-vivo animal models and where possible in human studies. Recognition of the coordinated regulation of cellular Na absorptive and Cl-secretory pathways together with the paracellular route in health and disease will help develop a more holistic picture of the multifaceted nature of small intestinal ion transport. PMID- 20010101 TI - The role of keratinocytes in defense against infection. AB - PURPOSE OF REVIEW: The discovery of increasing numbers of epithelial antimicrobial peptides (AMPs), cytokines that specifically induce AMPs in epithelial cells, and mechanisms of its regulation point toward a central role of the keratinocyte as effector cell of the epithelial innate defense system. This review summarizes recent progress in understanding the keratinocyte's role in combating infection that can help to understand why skin is usually covered with microbes but normally not infected. RECENT FINDINGS: The AMP LL-37 has been identified as regulator of keratinocyte apoptosis. The hypothesis of a direct defense function of keratinocytes, combating bacterial, fungal, virus and parasite infection, is strengthened. The discovery of an IL-22-producing T lymphocyte subpopulation implicates a role in AMP induction of keratinocytes. Multiple studies are adding to our understanding of how skin keratinocytes are interacting in skin barrier defects and with the microflora. Although in atopic patients AMP production is not generally impaired, in hyperIgE syndrome a lack of Th17 cytokines causes local Staphylococcus aureus infection due to a defective keratinocyte defense system. Ultraviolet radiation induces AMPs and thus may have beneficial effects to combat skin infection. SUMMARY: There is better understanding of the keratinocyte's role in the skin's innate defense system, and these data can help to generate therapeutics that can activate this defense system. PMID- 20010102 TI - HIV and bone mineral density. AB - PURPOSE OF REVIEW: This review details the clinical aspects and pathogenesis of low bone mineral density (BMD) in HIV, discusses broad management issues and outlines areas in which our understanding of this condition is incomplete. RECENT FINDINGS: Low BMD is prevalent in HIV-infected patients, with traditional risk factors, HIV infection and exposure to antiretroviral therapy all contributing. The role of specific antiretrovirals in the development of low BMD remains controversial, but most changes arise at either antiretroviral therapy initiation or switch. SUMMARY: Further research is needed to clarify mechanisms underlying low BMD in HIV, whether low BMD will translate to increased fractures and to determine the correct therapeutic approach to low BMD in HIV, particularly in younger HIV-infected patients. PMID- 20010103 TI - Surgical techniques for pancreas transplantation. AB - PURPOSE OF REVIEW: Pancreas transplantation reproducibly induces insulin independence in beta-cell penic diabetic patients. The difference between full insulin independence, partial graft function, and graft loss, mostly results from technical failure, graft rejection, and patient death with function graft. The purpose of this review is to examine recent surgical advances and discuss their contribution to improved graft function. RECENT FINDINGS: Few actual surgical innovations were described in the period reviewed. Duodenoduodenostomy is an interesting option for drainage of digestive secretions, when the pancreas is placed behind the right colon and is oriented cephalad. The main advantage of this technique is easy endoscopic assessment of donor duodenum but, when allograft pancreatectomy is necessary, repair of native duodenum may be troublesome. Selective revascularization of the gastroduodenal artery, at the back-table, possibly improves blood supply to the head of the pancreas graft and duodenal segment. There is no proof that this additional maneuver is always beneficial, although it can be graft saving in case of poor segmental graft perfusion. SUMMARY: Transplant surgeons should be familiar with all techniques for pancreas transplantation. Long-term graft function is possible only after technically successful pancreas transplantation. There is clearly a need for more objective assessment and standardization of surgical techniques for pancreas transplantation. PMID- 20010104 TI - Impact of pancreas transplantation on type 1 diabetes-related complications. AB - PURPOSE OF REVIEW: Type 1 diabetes is a chronic disease that can impact patient survival and quality of life because of acute and chronic complications. Although intensive insulin scheme treatment has been shown to reduce the incidence of diabetes-related complications, only pancreas transplantation has been shown to be able to alter them and in some cases to revert them. In this review, an extensive view of the effect of pancreas transplantation on diabetes-related complication will be described. RECENT FINDINGS: This review will focus on patients survival, diabetic nephropathy, neuropathy, cardiovascular event, comparing their incidence in type 1 diabetic patients treated with insulin and in type 1 diabetic patients receiving kidney, kidney-pancreas or pancreas alone graft. The review will focus mostly on the papers published in the last decade, with a particular attention to those on new aspects of graft function analysis like spectroscopy. Moreover, a comparison with islet transplantation procedure will be performed. SUMMARY: This review will give an update on the potential of pancreas transplantation, give a guide for clinical practice and help to consider pancreas transplantation as an alternative to insulin treatment for selected patients. PMID- 20010105 TI - Primary care management of chronic obstructive pulmonary disease: an integrated goal-directed approach. AB - PURPOSE OF REVIEW: This review discusses the role of the primary care physician in the care of the patient with chronic obstructive pulmonary disease and considers how an integrated chronic care model can be applied. RECENT FINDINGS: Evidence suggests that a multidisciplinary approach can be successfully applied to a chronic obstructive pulmonary disease patient. These interventions can lead to improvement in quality of life and reduced healthcare utilization. SUMMARY: An integrated care program for chronic obstructive pulmonary disease to assist the primary care provider should include the elements of the chronic care model. The support of a community-based outreach nurse, allied health staff, rehabilitation services and a case manager can assure best practices and identify gaps in quality care. PMID- 20010106 TI - The GlideScope for tracheal intubation in patients with grade IV modified Cormack and Lehane. PMID- 20010107 TI - Update on anaesthetic approach to pulmonary hypertension. AB - Pulmonary hypertensive patients going for surgery are at significant risk of perioperative morbidity and mortality. They pose a clinically challenging situation for the anaesthetist and strategies to minimize the effects on these patients are discussed. Recent advances have allowed a better understanding of the pathophysiological mechanisms and development of new therapies. We present a framework for preoperative assessment and optimization. An update on the recent advances in medical therapy with calcium channel blockers, prostacyclin analogues, endothelin receptor antagonists and phosphodiesterase-5 inhibitors is provided and relevant anaesthetic concerns highlighted. Key strategies for intraoperative management are presented and we review the options and novel strategies for managing an acute episode of decompensated pulmonary hypertension intraoperatively. Promising new therapies being explored include intraoperative delivery of inhaled milrinone and inhaled prostacyclin analogues. PMID- 20010108 TI - Long-term treatment of neuropathic pain with a 5% lidocaine medicated plaster. AB - BACKGROUND AND OBJECTIVE: The 5% lidocaine medicated plaster is a topical treatment for peripheral neuropathic pain symptoms (e.g. burning, shooting and stabbing pain) and is registered for the treatment of postherpetic neuralgia. This study examined the efficacy and tolerability of long-term treatment with the 5% lidocaine medicated plaster in patients with localized neuropathic pain conditions. METHODS: Twenty patients with localized neuropathic pain [postoperative neuropathic pain (n = 14); complex regional pain syndrome (n = 2); and postherpetic neuralgia (n = 4)], who had been successfully treated with 5% lidocaine medicated plaster, were followed up by telephone interview after 3 and 5 years. Questions were related to the efficacy, development of tolerance, tolerability, wear time and comfort of the plaster. RESULTS: At 3 years, 10 out of 20 (50%) initial responders were still using the plasters with no decline in analgesic efficacy. After 5 years, eight of the original 20 responders (40%) maintained treatment and continued to experience effective pain relief. The 12 responders who discontinued treatment did so because they no longer required analgesic therapy (n = 4); their health insurer refused to fund treatment (n = 2); they were lost to follow-up (n = 1); or had died from an illness unrelated to plaster treatment (n = 5). No patient discontinued because of inadequate analgesia or intolerable side effects. Reversible erythema occurred in two patients wearing the plaster for more than 16 h. There were no systemic side effects. CONCLUSION: The 5% lidocaine medicated plaster provides sustained pain relief over long-term treatment in patients with neuropathic pain of various causes and is well tolerated. PMID- 20010109 TI - Split right coronary artery. PMID- 20010110 TI - Trend in implantable cardioverter defibrillators and relation to need. AB - BACKGROUND: Rates of implantation of implantable cardioverter defibrillators (ICD) have increased continuously ever since they were first introduced. Notwithstanding guidelines endorsed by international scientific organizations, their use varies greatly across industrialized countries. The aim of this study was to assess whether variations in temporal trends and geographical distribution in the use of ICD devices across Piedmont, Northern Italy, are related to variations in need. METHODS: We calculated ICD implantation rates in the 19 local health units of Piedmont from 1999 to 2006, and correlated their temporal trend with four proxy indicators of need: coronary heart disease (CHD) mortality, CHD hospital discharge rates, cardiac heart failure discharge rates and bisoprolol or carvedilol prescription rates. RESULTS: The ICD implantation rate increased five fold between 1999-2000 and 2005-2006, mainly among the elderly. Implantation rates were five-fold higher in men compared to women for the entire duration of the study. There were significant differences between local health units, which increased over time. Among men there was only a statistically significant correlation with coronary heart disease mortality (r = 0.66) in the period from 2005 to 2006, and with the use of bisoprolol and carvedilol starting from 2001 to 2002. No significant correlation with need indicators was found in women. CONCLUSION: The use of ICD devices increased in apparent response to new research evidence, and, at least in part, in response to need. However, this process only involved men; ICD devices are largely underused in women and without apparent relation to need. PMID- 20010111 TI - Cardiac adrenergic nerve function in patients with cardiac syndrome X. AB - BACKGROUND: We previously found a severe impairment of cardiac uptake of I metaiodobenzylguanidine (MIBG), an analogue of norepinephrine, on myocardial scintigraphy in a small group of patients with cardiac syndrome X (CSX), suggesting a dysfunction of cardiac adrenergic nerve fibres. In this study, we assessed the consistency of these previous findings in a larger group of these patients. METHODS: Planar and single-photon emission computed tomography MIBG myocardial scintigraphy was performed in 40 CSX patients (58 +/- 7 years, 17 men). Cardiac MIBG uptake was measured by the heart/mediastinum ratio and by a single-photon emission computed tomography regional cardiac MIBG uptake defect score (higher values = lower uptake). As a control group, we studied 20 healthy individuals (56 +/- 6 years, nine men). An exercise stress Tc-SestaMIBI myocardial scintigraphy was performed in 34 CSX patients (85%). RESULTS: Cardiac MIBG defects were observed in 30 patients (75%), with nine (22.5%) showing no cardiac MIBG uptake at all. Compared with controls, CSX patients showed a significantly lower heart/mediastinum ratio (1.70 +/- 0.35 vs. 2.1 +/- 0.22, P < 0.001) and a higher cardiac MIBG defect score (27 +/- 25 vs. 4.4 +/- 2.5, P < 0.001). No differences were found in lung MIBG uptake between the two groups. Reversible perfusion defects on stress myocardial scintigraphy were found in 17 out of 34 CSX patients (50%), all of whom also had abnormal cardiac MIBG uptake; cardiac MIBG uptake abnormalities were also present in nine of 17 patients with normal perfusion scintigraphic images. Cardiac MIBG uptake findings were similar in our first 12 patients and in the 28 patients studied subsequently. CONCLUSION: Our data show a relevant impairment of cardiac MIBG uptake in patients with CSX, suggesting that functional abnormalities in cardiac adrenergic nerve function may play a significant role in the mechanisms responsible for the syndrome. PMID- 20010112 TI - Muscle contractile function and neural control after repetitive endurance cycling. AB - PURPOSE: To examine alterations in muscle contractile properties, cortical excitability, and voluntary activation as a consequence of 20 d of repetitive endurance cycling within a 22-d period. METHODS: Eight well-trained male cyclists completed 20 prolonged cycling stages interspersed by two rest days (days 9 and 17), which replicated the 2007 Tour de France route and schedule. Isometric knee extensor torque and EMG responses of the vastus lateralis in response to percutaneous electrical stimulation and transcranial magnetic stimulation were measured before, on days 9 and 17, and 2 d after completion of Tour de France. Postexercise measurements on days 9 and 17 were taken >18 h after cessation of the previous exercise bout. RESULTS: Maximal voluntary contraction of the knee extensors decreased by 20 +/- 10% (P < 0.01) during Tour de France but recovered after 2 d of rest. Peripherally evoked M-wave and potentiated twitch responses were also significantly decreased during Tour de France, up to 31 +/- 21% and 22 +/- 18%, respectively (P < 0.05), but returned to baseline values after 2 d of recovery. Voluntary activation was reduced to 75 +/- 8% (P < 0.05) during Tour de France and remained significantly depressed (79 +/- 7%, P < 0.05) after completion. The amplitude of motor evoked potentials was decreased by 44 +/- 28% (P < 0.01) on day 9 and remained significantly depressed during the remainder of, and after, Tour de France. CONCLUSIONS: A reduction in knee extensor strength, which occurs after repetitive prolonged cycling exercise, is a result of both central and peripheral processes. Reduced sarcolemmal excitability and impairment of contractile mechanisms exists even after 18 h of recovery. An enduring reduction in corticomotor output persists even after 2 d of rest. PMID- 20010113 TI - Gait termination control strategies are altered in chronic ankle instability subjects. AB - Despite the high incidence of chronic ankle instability (CAI), the underlying neurophysiologic mechanism is unknown. Evidence suggests that both feed-forward and feedback mechanisms may play a role. However, no investigation has examined both control mechanisms during the same movement task in the same cohort of CAI patients. PURPOSE: To determine the neuromuscular and biomechanical control alterations present in CAI patients during planned (feed-forward) and unplanned (feedback) gait termination. METHODS: Twenty subjects with CAI and 20 uninjured controls completed planned and unplanned gait termination protocols. Both tasks began with subjects walking at a self-selected speed across a 12-m walkway. Unplanned gait termination required subjects to stop during randomly selected trials on two adjacent force plates when cued. Planned gait termination required purposeful stopping on the force places. Propulsive and braking force magnitude and the dynamic postural stability index were calculated from the resulting ground reaction forces. In addition, muscle activity from the soleus, tibialis anterior, and gluteus medius was collected bilaterally. RESULTS: Both maximum propulsive (CAI = 99.8 +/- 40.8 N, control = 88.6 +/- 33.6 N) and braking (CAI = 207.1 +/- 80.9 N, control = 161.6 +/- 62.2 N) forces were significantly higher in the CAI group. The dynamic postural stability index revealed higher scores in the CAI group (0.24 +/- 0.03) compared with the control group (0.22 +/- 0.03). Muscle activation of the soleus and tibialis anterior differed during unplanned and planned gait termination between groups (P < 0.05) and between the limbs of the CAI group (P < 0.05). CONCLUSIONS: Altered biomechanical strategies during both planned and unplanned gait termination indicate that patients with CAI have alterations in feed-forward neuromuscular control and suggest the presence of feedback neuromuscular control deficits. PMID- 20010114 TI - Double-push skating versus V2 and V1 skating on uphill terrain in cross-country skiing. AB - PURPOSE: The aims of the study were a) to compare the double-push skating technique with the V2 and the V1 skating techniques on an uphill terrain by a kinematic and kinetic analysis, b) to provide kinetic and kinematic data of the V1 technique at maximal skiing speeds, and c) to test the hypotheses that the double-push skating technique is faster compared with the V2 and the V1 skating techniques. METHODS: Six elite skiers performed maximum speed sprints over a 60-m uphill section (7 degrees -10 degrees) using the double-push, the V2, and the V1 techniques. Pole and plantar forces and cycle characteristics were analyzed. RESULTS: The double-push skating technique was approximately 4.3% faster (P < 0.05) compared with the V2 skating technique and equally fast compared with the V1 skating technique. The double-push and the V2 techniques demonstrated longer cycle lengths, lower cycle rates (both P < 0.05), and equal poling frequencies and pole forces compared with the V1 technique. Cycle length, peak foot force, and knee extension ranges of motion and velocities were higher in the double-push technique compared with the V2 technique (all P values <0.05). Center of pressure was located more laterally in the double-push technique compared with the other two techniques (P < 0.05). All measured skiing speeds were drastically higher compared with former studies. CONCLUSION: The higher skiing speeds of the V1 and the double-push techniques compared with the V2 technique stress the mechanical advantage of those techniques on uphill terrain. Because of larger cycle lengths, lower cycle rate, longer recovery times, and equal poling frequency, the double push technique might be seen as more economic on steep uphills compared with the V1 technique. From a tactical point of view compared with the V1 technique, the double-push technique needs less space due to less lateral displacement, and no technique transitions are necessary when entering and leaving an uphill section. PMID- 20010115 TI - Predicting VO2max with an objectively measured physical activity in Japanese women. AB - PURPOSE: To investigate the use of the accelerometer-determined physical activity (PA) intensity variables as the objective PA variables for estimating VO2max in Japanese adult women. METHODS: The subjects of this study were 148 Japanese women aged 20 to 69 yr. Maximal oxygen uptake (VO2max) was measured with a maximal incremental test on a bicycle ergometer. Daily step counts (SC) and the amount spent in moderate to vigorous PA (MVPA) and vigorous PA (VPA) were measured using accelerometer-based activity monitors for 7 consecutive days. Using data of age, SC, MVPA, or VPA, and either body mass index (BMI) or waist circumference (WC), the nonexercise VO2max prediction models were derived as BMI models(MVPA), WC models(MVPA), BMI models(VPA), and WC models(VPA), and cross-validated by using two separate cross-validation procedures. RESULTS: SC, MVPA, and VPA were significantly related to VO2max (r = 0.43, r = 0.52, and r = 0.58, respectively). The multiple correlation coefficients for the BMI and WC models(MVAP) were 0.83 and 0.85, respectively, and for the BMI and WC models(VPA), they were 0.85 and 0.86, respectively. The SEE was 3.3 and 3.1 mL x kg(-1) x min(-1) for the BMI and WC models(MVPA), respectively, and it was 3.1 and 3.0 mL x kg(-1) x min(-1) for the BMI and WC models(VPA), respectively. All regression models demonstrated a high level of cross-validity supported by the minor shrinkage of the coefficient of determination and the increment of SEE in the predicted residual sum of squares procedure, and by small constant errors for the subgroups of age, SC, and VO2max between 25 and 35 mL x kg(-1) x min(-1). CONCLUSIONS: This study demonstrated that multiple regression models using data of MVPA or VPA were useful in predicting VO2max for Japanese adult women. PMID- 20010116 TI - Energy cost and metabolic power in elite soccer: a new match analysis approach. AB - PURPOSE: Video match analysis is used for the assessment of physical performances of professional soccer players, particularly for the identification of "high intensities" considered as "high running speeds." However, accelerations are also essential elements setting metabolic loads, even when speed is low. We propose a more detailed assessment of soccer players' metabolic demands by video match analysis with the aim of also taking into account accelerations. METHODS: A recent study showed that accelerated running on a flat terrain is equivalent to running uphill at constant speed, the incline being dictated by the acceleration. Because the energy cost of running uphill is known, this makes it possible to estimate the instantaneous energy cost of accelerated running, the corresponding instantaneous metabolic power, and the overall energy expenditure, provided that the speed (and acceleration) is known. Furthermore, the introduction of individual parameters makes it possible to customize performance profiles, especially as it concerns energy expenditure derived from anaerobic sources. Data from 399 "Serie-A" players (mean +/- SD; age = 27 +/- 4 yr, mass = 75.8 +/- 5.0 kg, stature = 1.80 +/- 0.06 m) were collected during the 2007-2008 season. RESULTS: Mean match distance was 10,950 +/- 1044 m, and average energy expenditure was 61.12 +/- 6.57 kJ x kg(-1). Total distance covered at high power (>20 W x kg(-1)) amounted to 26% and corresponding energy expenditure to approximately 42% of the total. "High intensities" expressed as high-power output are two to three times larger than those based only on running speed. CONCLUSIONS: The present approach for the assessment of top-level soccer players match performance through video analysis allowed us to assess instantaneous metabolic power, thus redefining the concept of "high intensity" on the basis of actual metabolic power rather than on speed alone. PMID- 20010117 TI - Spontaneous pacing during overground hill running. AB - PURPOSE: To investigate speed regulation during overground running on undulating terrain. METHODS: After an initial laboratory session to calculate physiological thresholds, eight experienced runners completed a spontaneously paced time trial over three laps of an outdoor course involving uphill, downhill, and level sections. A portable gas analyzer, global positioning system receiver, and activity monitor were used to collect physiological, speed, and stride frequency data. RESULTS: Participants ran 23% slower on uphills and 13.8% faster on downhills compared with level sections. Speeds on level sections were significantly different for 78.4 +/- 7.0 s following an uphill and 23.6 +/- 2.2 s following a downhill. Speed changes were primarily regulated by stride length, which was 20.5% shorter uphill and 16.2% longer downhill, whereas stride frequency was relatively stable. Oxygen consumption averaged 100.4% of runner's individual ventilatory thresholds on uphills, 78.9% on downhills, and 89.3% on level sections. Approximately 89% of group-level speed was predicted using a modified gradient factor. Individuals adopted distinct pacing strategies, both across laps and as a function of gradient. CONCLUSIONS: Speed was best predicted using a weighted factor to account for prior and current gradients. Oxygen consumption (VO2) limited runner's speeds only on uphill sections and was maintained in line with individual ventilatory thresholds. Running speed showed larger individual variation on downhill sections, whereas speed on the level was systematically influenced by the preceding gradient. Runners who varied their pace more as a function of gradient showed a more consistent level of oxygen consumption. These results suggest that optimizing time on the level sections after hills offers the greatest potential to minimize overall time when running over undulating terrain. PMID- 20010118 TI - Caloric restriction with or without exercise: the fitness versus fatness debate. AB - There is a debate over the independent effects of aerobic fitness and body fatness on mortality and disease risks. PURPOSE: To determine whether a 25% energy deficit that produces equal change in body fatness leads to greater cardiometabolic benefits when aerobic exercise is included. METHODS: Thirty-six overweight participants (16 males/20 females) (39 +/- 1 yr; 82 +/- 2 kg; body mass index = 27.8 +/- 0.3 kg x m2, mean +/- SEM) were randomized to one of three groups (n = 12 for each) for a 6-month intervention: control (CO, weight maintenance diet), caloric restriction (CR, 25% reduction in energy intake), or caloric restriction plus aerobic exercise (CR + EX, 12.5% reduction in energy intake plus 12.5% increase in exercise energy expenditure). Food was provided during weeks 1-12 and 22-24. Changes in fat mass, visceral fat, VO2peak (graded treadmill test), muscular strength (isokinetic knee extension/flexion), blood lipids, blood pressure, and insulin sensitivity/secretion were compared. RESULTS: As expected, VO2peak was significantly improved after 6 months of intervention in CR + EX only (22 +/- 5% vs 7 +/- 5% in CR and -5 +/- 3% in CO), whereas isokinetic muscular strength did not change. There was no difference in the losses of weight, fat mass, or visceral fat and changes in systolic blood pressure (BP) between the intervention groups. However, only CR + EX had a significant decrease in diastolic BP (-5 +/- 3% vs -2 +/- 2% in CR and -1 +/- 2% in CO), in low-density lipoprotein (LDL) cholesterol (-13 +/- 4% vs -6 +/- 3% in CR and 2 +/- 4% in CO), and a significant increase in insulin sensitivity (66 +/- 22% vs 40 +/- 20% in CR and 1 +/- 11% in CO). CONCLUSIONS: Despite similar effect on fat losses, combining CR with exercise increased aerobic fitness in parallel with improved insulin sensitivity, LDL cholesterol, and diastolic BP. The results lend support for inclusion of an exercise component in weight loss programs to improve metabolic fitness. PMID- 20010119 TI - Ergogenic and antioxidant effects of spirulina supplementation in humans. AB - PURPOSE: Spirulina is a popular nutritional supplement that is accompanied by claiMSS for antioxidant and performance-enhancing effects. Therefore, the aim of the present study was to examine the effect of spirulina supplementation on (i) exercise performance, (ii) substrate metabolism, and (iii) blood redox status both at rest and after exercise. METHODS: Nine moderately trained males took part in a double-blind, placebo-controlled, counterbalanced crossover study. Each subject received either spirulina (6 g x d(-1)) or placebo for 4 wk. Each subject ran on a treadmill at an intensity corresponding to 70%-75% of their VO2max for 2 h and then at 95% VO2max to exhaustion. Exercise performance and respiratory quotient during exercise were measured after both placebo and spirulina supplementation. Blood samples were drawn before, immediately after, and at 1, 24, and 48 h after exercise. Reduced glutathione (GSH), oxidized glutathione (GSSG), GSH/GSSG, thiobarbituric acid-reactive substances (TBARS), protein carbonyls, catalase activity, and total antioxidant capacity (TAC) were determined. RESULTS: Time to fatigue after the 2-h run was significantly longer after spirulina supplementation (2.05 +/- 0.68 vs 2.70 +/- 0.79 min). Ingestion of spirulina significantly decreased carbohydrate oxidation rate by 10.3% and increased fat oxidation rate by 10.9% during the 2-h run compared with the placebo trial. GSH levels were higher after the spirulina supplementation compared with placebo at rest and 24 h after exercise. TBARS levels increased after exercise after placebo but not after spirulina supplementation. Protein carbonyls, catalase, and TAC levels increased similarly immediately after and 1 h after exercise in both groups. CONCLUSIONS: Spirulina supplementation induced a significant increase in exercise performance, fat oxidation, and GSH concentration and attenuated the exercise-induced increase in lipid peroxidation. PMID- 20010120 TI - Aerobic performance is degraded, despite modest hyperthermia, in hot environments. AB - Environmental heat stress degrades aerobic performance; however, little research has focused on performance when the selected task elicits modest elevations in core body temperature (<38.5 degrees C). PURPOSE: To determine the effect of environmental heat stress, with modest hyperthermia, on aerobic performance and pacing strategies. METHODS: After a 30-min cycling preload at 50% VO2peak, eight euhydrated men performed a 15-min time trial on a cycle ergometer in temperate (TEMP; 21 degrees C, 50% RH) and hot (HOT; 40 degrees C, 25% RH) environments. Core and skin temperature (Tc and Tsk, respectively) and HR were continuously monitored. Performance was assessed by the total work (kJ) completed in 15 min. Pacing was quantified by comparing the percent difference in actual work performed in each of five 3-min blocks normalized to the mean work performed per 3-min block. Pace over the final 2 min was compared with the average pace from minutes 0 to 13 for end spurt analysis. RESULTS: Tc and HR rose continually throughout both time trials. Peak Tc remained modestly elevated in both environments [mean (range): HOT = 38.20 degrees C (37.97-38.42 degrees C); TEMP = 38.11 degrees C (38.07-38.24 degrees C)], whereas Tsk was higher in HOT (36.19 +/ 0.40 degrees C vs 31.14 +/- 1.14 degrees C), and final HR reached approximately 95% of age-predicted maximum in both environments. Total work performed in HOT (147.7 +/- 23.9 kJ) was approximately 17% less (P < 0.05) than TEMP (177.0 +/- 25.0 kJ). Pace was evenly maintained in TEMP, but in HOT, volunteers were unable to maintain initial pace, slowing progressively over time. A significant end spurt was produced in both environments. CONCLUSIONS: During a brief aerobic exercise time trial where excessive hyperthermia is avoided, total work is significantly reduced by heat stress because of a gradual slowing of pace over time. These findings demonstrate how aerobic exercise performance degrades in hot environments without marked hyperthermia. PMID- 20010121 TI - Faster pulmonary oxygen uptake kinetics in trained versus untrained male adolescents. AB - Exercise training results in a speeding of pulmonary oxygen uptake (VO2) kinetics at the onset of exercise in adults; however, only limited research has been conducted with children and adolescents. PURPOSE: The aim of the present study was to examine VO2 and muscle deoxygenation kinetics in trained and untrained male adolescents. METHODS: Sixteen trained (15 +/- 0.8 yr, VO2peak = 54.7 +/- 6.2 mL x kg-1 x min-1, self-assessed Tanner stage range 2-4) and nine untrained (15 +/- 0.6 yr, VO2peak = 43.1 +/- 5.2 mL x kg-1 x min-1, Tanner stage range 2-4) male adolescents performed two 6-min exercise transitions from a 3-min baseline of 10 W to a workload equivalent to 80% lactate threshold separated by a minimum of 1 h of passive rest. Oxygen uptake (breath-by-breath) and muscle deoxygenation (deoxyhemoglobin signal from near-infrared spectroscopy) were measured continuously throughout baseline and exercise transition. RESULTS: The time constant of the fundamental phase of VO2 kinetics was significantly faster in trained versus untrained subjects (trained: 22.3 +/- 7.2 s vs untrained: 29.8 +/- 8.4 s, P = 0.03). In contrast, neither the time constant (trained: 9.7 +/- 2.9 s vs untrained: 10.1 +/- 3.4 s, P = 0.78) nor the mean response time (trained: 17.4 +/- 2.5 s vs untrained: 18.3 +/- 2.3 s, P = 0.39) of muscle deoxygenation kinetics differed with training status. CONCLUSIONS: The present data suggest that exercise training results in faster VO2 kinetics in male adolescents, although inherent capabilities cannot be ruled out. Because muscle deoxygenation kinetics were unchanged, it is likely that faster VO2 kinetics were due to adaptations to both the cardiovascular system and the peripheral musculature. PMID- 20010122 TI - Exercise-induced hypoxemia: fact or fallacy? AB - Whereas the prevalence of exercise-induced hypoxemia (EIH) in endurance athletes is commonly reported as approximately 50%, most previous studies have not corrected PaO2 for exercise-induced hyperthermia. Furthermore, although a detrimental effect on aerobic performance has been assumed, no study has measured arterial oxygen content (CaO2) in this context. PURPOSE: To determine the effect of temperature-correcting PaO2 values for rectal, arterial blood, esophageal, and exercising muscle temperatures during exercise on CaO2 and the prevalence of EIH. METHODS: Twenty-three trained males (age 26 +/- 5 yr; VO2peak 65.2 +/- 1.6 mL x kg-1 x min-1) performed incremental treadmill exercise to exhaustion with PaO2 corrected for simultaneous temperature measurements at all four sites. EIH was defined as DeltaPaO2 >or= 10 mm Hg. RESULTS: : With no temperature correction, DeltaPaO2 was -20.8 +/- 5.0 mm Hg and prevalence was 96% (n = 23), but when corrected for rectal temperature, DeltaPaO2 was -14.7 +/- 7.8 mm Hg and prevalence was 73% (n = 20); for arterial blood temperature, DeltaPaO2 was -7.7 +/- 6.5 mm Hg and prevalence was 35% (n = 20); and for esophageal temperature, DeltaPaO2 was -8.1 +/- 7.7 mm Hg and prevalence was 48% (n = 23), although when corrected for active muscle temperature, DeltaPaO2 was +8.2 +/- 7.8 mm Hg and prevalence was 0% (n = 10). There were no significant changes in CaO2 except for uncorrected values, and there was no correlation between DeltaPaO2 and VO2peak. CONCLUSIONS: Although the prevalence of EIH depends on the temperature correction applied to PaO2 values, in no case is there a significant change in CaO2 or any relationship with maximal aerobic power. PMID- 20010123 TI - Low-intensity swimming training partially inhibits lipopolysaccharide-induced acute lung injury. AB - BACKGROUND: Aerobic exercise decreases pulmonary inflammation and remodeling in experimental models of allergic asthma. However, the effects of aerobic exercise on pulmonary inflammation of nonallergic origin, such as in experimental models of acute lung injury induced by lipopolysaccharide (LPS), have not been evaluated. OBJECTIVE: The present study evaluated the effects of aerobic exercise in a model of LPS-induced acute lung injury. METHODS: BALB/c mice were divided into four groups: Control, Aerobic Exercise, LPS, and Aerobic Exercise + LPS. Swimming tests were conducted at baseline and at 3 and 6 wk. Low-intensity swimming training was performed for 6 wk, four times per week, 60 min per session. Intranasal LPS (1 mg x kg(-1) (60 microg per mouse)) was instilled 24 h after the last swimming physical test in the LPS and Aerobic Exercise + LPS mice, and the animals were studied 24 h after LPS instillation. Exhaled nitric oxide, respiratory mechanics, total and differential cell counts in bronchoalveolar lavage, and lung parenchymal inflammation and remodeling were evaluated. RESULTS: LPS instillation resulted in increased levels of exhaled nitric oxide (P < 0.001), higher numbers of neutrophils in the bronchoalveolar lavage (P < 0.001) and in the lung parenchyma (P < 0.001), and decreased lung tissue resistance (P < 0.05) and volume proportion of elastic fibers (P < 0.01) compared with the Control group. Swim training in LPS-instilled animals resulted in significantly lower exhaled nitric oxide levels (P < 0.001) and fewer neutrophils in the bronchoalveolar lavage (P < 0.001) and the lung parenchyma (P < 0.01) compared with the LPS group. CONCLUSIONS: These results suggest that low-intensity swimming training inhibits lung neutrophilic inflammation, but not remodeling and impaired lung mechanics, in a model of LPS-induced acute lung injury. PMID- 20010124 TI - ACTN3 and ACE genotypes in elite Jamaican and US sprinters. AB - The angiotensin-converting enzyme (ACE) and the alpha-actinin-3 (ACTN3) genes are two of the most studied "performance genes" and both have been associated with sprint/power phenotypes and elite performance. PURPOSE: To investigate the association between the ACE and the ACTN3 genotypes and sprint athlete status in elite Jamaican and US African American sprinters. METHODS: The ACTN3 R577X and the ACE I/D and A22982G (rs4363) genotype distributions of elite Jamaican (J-A; N = 116) and US sprinters (US-A; N = 114) were compared with controls from the Jamaican (J-C; N = 311) and US African American (US-C; N = 191) populations. Frequency differences between groups were assessed by exact test. RESULTS: For ACTN3, the XX genotype was found to be at very low frequency in both athlete and control cohorts (J-C = 2%, J-A = 3%, US-C = 4%, US-A = 2%). Athletes did not differ from controls in ACTN3 genotype distribution (J, P = 0.87; US, P = 0.58). Similarly, neither US nor Jamaican athletes differed from controls in genotype at ACE I/D (J, P = 0.44; US, P = 0.37). Jamaican athletes did not differ from controls for A22982G genotype (P = 0.28), although US sprinters did (P = 0.029), displaying an excess of heterozygotes relative to controls but no excess of GG homozygotes (US-C = 22%, US-A = 18%). CONCLUSIONS: Given that ACTN3 XX genotype is negatively associated with elite sprint athlete status, the underlying low frequency in these populations eliminates the possibility of replicating this association in Jamaican and US African American sprinters. The finding of no excess in ACE DD or GG genotypes in elite sprint athletes relative to controls suggests that ACE genotype is not a determinant of elite sprint athlete status. PMID- 20010125 TI - Cycle training increased GLUT4 and activation of mammalian target of rapamycin in fast twitch muscle fibers. AB - PURPOSE: To determine whether cycle training of sedentary subjects would increase the expression of the principle muscle glucose transporters, six volunteers completed 6 wk of progressively increasing intensity stationary cycle cycling. METHODS: In vastus lateralis muscle biopsies, changes in expression of GLUT1, GLUT4, GLUT5, and GLUT12 were compared using quantitative immunoblots with specific protein standards. Regulatory pathway components were evaluated by immunoblots of muscle homogenates and immunohistochemistry of microscopic sections. RESULTS: GLUT1 was unchanged, GLUT4 increased 66%, GLUT12 increased 104%, and GLUT5 decreased 72%. A mitochondrial marker (cytochrome c) and regulators of mitochondrial biogenesis (peroxisome proliferator-activated receptor gamma coactivator 1 alpha and phospho-5'-adenosine monophosphate activated protein kinase) were unchanged, but the muscle hypertrophy pathway component, phospho-mammalian target of rapamycin (mTOR), increased 83% after the exercise program. In baseline biopsies, GLUT4 by immunohistochemical techniques was 37% greater in Type I (slow twitch, red) muscle fibers, but the exercise training increased GLUT4 expression in Type II (fast twitch, white) fibers by 50%, achieving parity with the Type I fibers. Baseline phospho-mTOR expression was 50% higher in Type II fibers and increased more in Type II fibers (62%) with training but also increased in Type I fibers (34%). CONCLUSION: Progressive intensity stationary cycle training of previously sedentary subjects increased muscle insulin-responsive glucose transporters (GLUT4 and GLUT12) and decreased the fructose transporter (GLUT5). The increase in GLUT4 occurred primarily in Type II muscle fibers, and this coincided with activation of the mTOR muscle hypertrophy pathway. There was little impact on Type I fiber GLUT4 expression and no evidence of change in mitochondrial biogenesis. PMID- 20010126 TI - Changes in calpain activity, muscle structure, and function after eccentric exercise. AB - PURPOSE: The aim of this study was to investigate changes in muscle function, muscle structure, and calpain activity after high-force eccentric exercise. METHODS: Eleven healthy males performed 300 maximal voluntary eccentric actions with knee extensors in one leg. Maximal force-generating capacity was measured before exercise and regularly during the next 7 d. Biopsies from musculus vastus lateralis were taken in both control and exercised legs 0.5, 4, 8, 24, 96, and 168 h after exercise for evaluation of myofibrillar structure, extracellular matrix proteins, and calpain activity. RESULTS: In the exercised leg, peak torque was reduced by 47 +/- 5% during exercise and was still 22 +/- 5% lower than baseline 4 d after the exercise. Calpain activity was three times higher in the exercised leg compared with the control leg 30 min after exercise. Myofibrillar disruptions were observed in 36 +/- 6% of all fibers in exercised muscle and in 2 +/- 1% of fibers in control muscle. The individual reductions in peak torque correlated with the proportion of fibers with myofibrillar disruptions (r = 0.89). The increase in calpain activity was not correlated to the proportion of fibers with myofibrillar disruptions. Nevertheless, the characteristics of the myofibrillar disruptions mimicked calpain-mediated degradation of myofibrils. Tenascin-C and the N-terminal propeptide of procollagen type III showed increased staining intensity on cross-sections 4-7 d after the exercise. CONCLUSIONS: Myofibrillar disruptions seem to be a main cause for the long-lasting reduction in force-generating capacity after high-force eccentric exercise. The increase in calpain activity, but the lack of a relationship between calpain activity and the amount of muscle damage, suggests multiple roles of calpain in the damage and repair process. PMID- 20010127 TI - Time course of leukocyte accumulation in human muscle after eccentric exercise. AB - PURPOSE: To investigate the time course of leukocyte accumulation in eccentric exercised human muscles and its relation to recovery of muscle function and soreness. METHODS: Eleven young males performed 300 unilateral, maximal voluntary, eccentric actions with the musculus quadriceps femoris (30 degrees x s(-1)). Before and at regular intervals for 7 d after exercise, force-generating capacity was measured with maximal concentric knee extensions (60 degrees x s( 1)). Accumulation of radiolabeled (autologous) leukocytes was measured with scintigraphy. Biopsies from musculus vastus lateralis were obtained 0.5, 4, 8, 24, 96, and 168 h after exercise from both the exercised leg and the control leg. Muscle cross-sections were stained with antibodies against leukocytes (CD16 and CD68). Muscle soreness was rated on a visual analog scale. RESULTS: Immediately after exercise, the subjects' ability to generate force was reduced by 47 +/- 5%. Muscle function recovered slowly and was not fully restored after 1 wk. Radiolabeled leukocytes accumulated in the muscles during the first hour (3-24 h) after exercise, and leukocytes were at the same time observed histologically, primarily in the endomysium and perimysium. A part of the accumulated radiolabeled leukocytes appeared to be located within local blood vessels. The highest numbers of CD16(+) and CD68(+) cells were found 4 and 7 d after exercise. There was a positive correlation between accumulation of radiolabeled leukocytes and muscle weakness measured 1-3 d after exercise (r = 0.8, P < 0.05) and, surprisingly, a negative correlation between radiolabeled leukocyte accumulation and muscle soreness (r = -0.96, P < 0.01). CONCLUSION: Exercise-induced muscle damage initiated a rapid local inflammatory response that gradually increased over the next days. Halted recovery of muscle function was associated with local accumulation of leukocytes, whereas muscle soreness could not be explained by the presence of leukocytes. PMID- 20010128 TI - Age-related differences in exercise and quality of life among breast cancer survivors. AB - PURPOSE: Physical activity has become a focus of cancer recovery research because it has the potential to reduce treatment-related burden and to optimize health related quality of life (HRQoL). However, the potential for physical activity to influence recovery may be age dependent. This article describes physical activity levels and HRQoL among younger and older women after surgery for breast cancer and explores the correlates of physical inactivity. METHODS: A population-based sample of breast cancer patients (n = 287) diagnosed in South-East Queensland, Australia, were assessed once every 3 months, from 6 to 18 months postsurgery. The Functional Assessment of Cancer Therapy-Breast questionnaire and items from the Behavioral Risk Factor Surveillance System questionnaire were used to measure HRQoL and physical activity, respectively. Physical activity was assigned MET values and categorized as <3, 3 to 17.9, and 18+ MET x h x wk. Descriptive statistics, generalized linear models with age stratification (<50 vs 50+ yr), and logistic regression were used for analyses (P = 0.05, two-tailed). RESULTS: Younger women who engaged in >or=3 MET x h x wk of physical activity reported a higher HRQoL at 18 months compared with their more sedentary counterparts (P < 0.05). Older women reported similar HRQoL irrespective of activity level and consistently reported clinically higher HRQoL than younger women. Increasing age, being overweight or obese, and restricting use of the treated side at 6 months postsurgery increased the likelihood of sedentary behavior (odds ratio >or= 3, P < 0.05). CONCLUSIONS: Age influences the potential to observe HRQoL benefits related to physical activity participation. These results also provide relevant information for the design of exercise interventions for breast cancer survivors and highlight that some groups of women are at greater risk of long-term sedentary behavior. PMID- 20010129 TI - Exercise and the growth hormone-insulin-like growth factor axis. AB - Exercise is a robust physiological stimulator of the pituitary secretion of growth hormone (GH), and within approximately 15 min after the onset of exercise, plasma GH starts to increase. GH and its primary downstream mediator, insulin like growth factor I (IGF-I), play a critical role in formation, maintenance, and regeneration of skeletal muscles. Consequently, it seems logical to link the exercise-induced stimulation of GH with the hypertrophy observed in exercising muscles. GH stimulates circulating (endocrine) as well as locally produced (peripheral) IGF-I, which acts through autocrine/paracrine mechanisms. However, it remains to be clarified whether skeletal muscle hypertrophy after exercise is stimulated primarily by endocrine or paracrine/autocrine IGF-I. Early cross sectional studies have observed positive correlations between circulating IGF-I levels and GH secretion, respectively, and indices of fitness. However, longitudinal exercise studies have shown that it is possible to increase muscle strength, performance, and VO2max without concomitant and robust changes in circulating IGF-I, indicating that the effect of exercise on skeletal muscles is mediated via paracrine/autocrine IGF-I rather than endocrine IGF-I. So far, most exercise studies have investigated the concentration of immunoreactive IGF-I in serum or plasma, obtained after extraction of the IGF-binding proteins (i.e., total IGF-I). However, several of the newer exercise studies have included measurement of free IGF-I as well as bioactive IGF-I. The aim of this review was to discuss whether measurement of free and/or bioactive IGF-I have increased our knowledge on the processes that link exercise, muscle hypertrophy, and GH/IGF-I axis. Thus, the current review will discuss (i) the different IGF-I assay methodologies and (ii) the current literature on free, bioactive, and immunoreactive (total) IGF-I in exercising subjects. PMID- 20010130 TI - Insulin-like growth factor I signaling in skeletal muscle and the potential for cytokine interactions. AB - Recent research has demonstrated that intracellular signaling components associated with several proinflammatory cytokines have the potential to interact with signaling pathways that regulate anabolic processes in skeletal muscle. This presentation and the ensuing brief review are intended to present a selection of the potential interactions between these two critical processes. PMID- 20010131 TI - Insulin-like growth factor I as a biomarker of health, fitness, and training status. AB - Whereas there are many varied roles of insulin-like growth factor I (IGF-I), and it exists in different biocompartments, there is abundant scientific evidence demonstrating that IGF-I is an important metabolic biomarker associated with a variety of health- and exercise-related outcomes. In most cases (muscle, bone, tendon, body composition, and cognitive function), elevated IGF-I concentrations are considered beneficial; however, cancer remains a notable exception. Although the fact that both increased and decreased IGF-I concentrations can be considered as reflective of favorable and beneficial health outcomes and may seem as a paradox and even contradictory, it is important to emphasize that, in both cases, measured IGF-I concentrations do offer important insight into physiological processes. The precise and relative role of systemic versus locally produced IGF I in mediating the outcomes of physical activity is still not clearly delineated, but it does seem as though local IGF-I is consistently upregulated with both acute and chronic exercises; whereas in certain situations, circulating IGF-I may actually decrease. Although perhaps counterintuitive to the known anabolic role that IGF-I exerts, positive neuromuscular training adaptations can occur in the presence of decreases or no changes in circulating IGF-I. These observations, however, should not be interpreted to conclude that the role of circulating IGF-I lacks importance or relevance in contributing to enhanced musculoskeletal health as evidenced by the liver IGF-I-deficient mouse model. Because of the ubiquitous nature of IGF-I, prospective experimental approaches involving physical activity that can sample and measure IGF-I in the body's various biocompartments (i.e., blood, interstitial fluid, muscle) with the most biologically relevant assays are encouraged. We believe that such endeavors will provide greater understanding in the complex role that IGF-I possesses in mediating exercise-induced adaptations. PMID- 20010132 TI - Insulin-like growth factor-I, physical activity, and control of cellular anabolism. AB - The underlying mechanisms responsible for mediating the beneficial outcomes of exercise undoubtedly are many, but the insulin-like growth factor-I (IGF-I) system is emerging as an important and central hormonal axis that plays a significant role concerning cellular anabolism. This introductory article summarizes the intent and the content for papers presented as part of a 2008 American College of Sports Medicine national symposium entitled "Insulin-like Growth Factor-I, Physical Activity, and Control of Cellular Anabolism." The individual authors and their papers are as follows: Jan Frystyk authoring "The relationship between exercise and the growth hormone/insulin-like growth factor-I axis," Greg Adams authoring "IGF-I signaling in skeletal muscle and the potential for cytokine interactions," and Brad Nindl authoring "Insulin-like growth factor I as a biomarker of health, fitness, and training status." These papers focus on 1) different assay methodologies for IGF-I within the paradigm of exercise studies, 2) research demonstrating that intracellular signaling components associated with several proinflammatory cytokines have the potential to interact with anabolic signaling processes in skeletal muscle, and 3) an overview of IGF-I as a biomarker related to exercise training, muscle and bone remodeling, body composition, cognition, and cancer. When summed in total, the contribution that these papers will make will undoubtedly involve bringing attention to the vast regulatory complexity of the IGF-I system and will hopefully convince the reader that the IGF-I system warrants further detailed scientific inquiry to resolve many unanswered questions and paradoxical experimental findings. The IGF-I system remains one of the most intriguing and captivating marvels of human physiology that seems central in mediating numerous adaptations from physical activity. PMID- 20010133 TI - Effect of progressive resistance training on muscle performance after chronic stroke. AB - PURPOSE: This study investigated the effects of high-intensity progressive resistance training (PRT) and high-intensity cycling (cycling) on muscle performance and the time course of strength gains in a chronic stroke population. METHODS: Forty-eight individuals with chronic stroke sequelae (mean +/- SD; age = 63 +/- 9 yr, time since stroke = 57 +/- 54 months) were randomly allocated to one of four treatment groups: PRT + cycling, PRT + sham cycling, sham PRT + cycling, or sham PRT + sham cycling groups in a fully factorial clinical trial. Thirty exercise sessions were conducted over a 10- to 12-wk period. The main outcomes investigated were measures of unilateral muscle strength, peak power, and muscle endurance. RESULTS: Those undergoing PRT improved their lower limb muscle strength, peak power, and endurance compared with participants receiving sham PRT or cycling only (P < 0.05), and combined exercise was not superior to PRT alone. Strength improvements occurred primarily during the first 6 to 8 wk (98%-100% of total gain) and then reached a plateau during the final 2 to 4 wk. CONCLUSION: We have shown for the first time in a direct comparison study that high-intensity PRT, but not cycling or sham exercise, can improve muscle strength, peak power, and muscle endurance in both affected and unaffected lower limbs after chronic stroke by a significant and clinically meaningful amount. Although strength gains plateaued earlier than anticipated, adherence to the intended continuous high intensity progressive overload protocol was largely achieved (average load of 84% +/- 4% of one repetition maximum). PMID- 20010134 TI - Sleep is not disrupted by exercise in patients with chronic fatigue syndromes. AB - PURPOSE: Patients with chronic fatigue syndrome (CFS) report that exertion produces dramatic symptom worsening. We hypothesized this might be due to the exacerbation of an underlying sleep disorder, which we have previously demonstrated to exist. METHODS: Female patients with CFS and matched healthy controls with no evidence of major depressive disorder were studied with overnight polysomnography on a baseline night and on a night after their performance of a maximal exercise test. RESULTS: CFS patients as a group had evidence for disturbed sleep compared with controls. Although exercise improved sleep for healthy subjects, it did not do this for the group as a whole. When we stratified the sample on the basis of self-reported sleepiness after a night's sleep, the patient group with reduced morning sleepiness showed improvement in sleep structure, whereas those with increased morning sleepiness continued to show evidence for sleep disruption. CONCLUSIONS: Sleep is disturbed in CFS patients as a group, but exercise does not exacerbate this sleep disturbance. Approximately half the patients studied actually sleep better after exercise. Therefore, activity-related symptom worsening is not caused by worsened sleep. PMID- 20010135 TI - Community-based aquatic exercise and quality of life in persons with osteoarthritis. AB - PURPOSE: To evaluate the effectiveness of a community-based aquatic exercise program for improved quality of life among persons with osteoarthritis. METHOD: Two hundred forty-nine adults with osteoarthritis were enrolled in a 20-wk randomized controlled trial of a preexisting community-based aquatic exercise program versus control. Intervention group participants (n = 125) were asked to attend at least two aquatic exercise sessions per week. Control group participants (n = 124) were asked to maintain their usual activity levels. Demographics were collected at baseline, and patient-reported outcomes were collected at baseline and after 10 and 20 wk. Depressive symptoms, self-efficacy for pain and symptom control, physical impairment, and activity limitation were tested as potential mediators of the relationship between aquatic exercise and perceived quality of life (PQOL). Body mass index (BMI), ethnicity, self-rated health, and comorbidity were tested as possible moderators. RESULTS: Aquatic exercise had a positive impact on PQOL scores (P < 0.01). This effect was moderated by BMI (P < 0.05) such that benefits were observed among obese participants (BMI >or= 30), but not among normal weight or overweight participants. None of the tested variables were found to mediate the relationship between aquatic exercise and PQOL scores. CONCLUSIONS: Given the availability of existing community aquatics programs, aquatic exercise offers a therapeutic and pragmatic option to promote quality of life among individuals who are living with both obesity and osteoarthritis. Future investigation is needed to replicate these findings and develop strategies to increase long-term participation in aquatics programs. PMID- 20010136 TI - Sickle cell trait and fatal rhabdomyolysis in football training: a case study. AB - We report the athletic, the clinical, and the pathological details of a case of fatal rhabdomyolysis during training in a college football player with sickle cell trait (SCT) who collapsed minutes after running 16 successive sprints of 100 yd each. The player, 19 yr old, African American, was apparently healthy when he took the field for the conditioning run. No exertional heat illness was present. After collapsing on-field, the player soon went into coma and developed fulminant rhabdomyolysis, profound lactic acidosis, acute myoglobinuric renal failure, refractory hyperkalemia, and disseminated intravascular coagulation. Despite intensive care in the hospital, he died about 15 h after admission, likely from a hyperkalemic cardiac arrhythmia; the terminal rhythm was pulseless electrical activity. The forensic autopsy confirmed that the cause of death was acute exertional rhabdomyolysis associated with SCT. Counting this case, at least 15 college football players with SCT have died from complications of exertional sickling, as have younger football players and other athletes. In SCT, maximal, sustained exercise evokes four forces that can foster sickling: hypoxemia, acidosis, hyperthermia, and red cell dehydration. The setting, the clinical and laboratory features, and the clinicopathological correlation here suggest that the fulminant rhabdomyolysis and its fatal sequelae were from exertional sickling. These data suggest that screening and simple precautions for SCT may be warranted to prevent tragedies like this and enable all athletes with SCT to thrive in their sports. PMID- 20010137 TI - Molecular mechanisms of keratinizing ocular surface disease. AB - A devastating consequence of autoimmune-mediated, aqueous tear deficiency is pathological keratinization of the ocular surface. It is setoff by an aberrant immune response that promotes a program of altered mucosal epithelial cell differentiation. The management of keratinizing ocular surface disease is challenging. Topical therapies are largely inadequate for acute exacerbations, and progressive disease often requires systemic immunosuppression. A combination of translational and basic science research is necessary to understand the link between aberrant immunity and pathological keratinization. I review recent research and future directions aimed to develop targeted therapies that control or prevent ocular surface keratinization. PMID- 20010138 TI - Enhancing visual performance for people with central vision loss. AB - People with central vision loss must use peripheral vision for visual tasks. It is well known that performance for almost all spatial tasks is worse in the normal periphery than in the normal fovea. The primary goal of my ongoing research is to understand the limiting factors and the potential for enhancing vision for people with central vision loss. Here I review my previous work related to understanding the limiting factors on reading, a task that is the primary complaint of many patients with age-related macular degeneration, the leading cause of visual impairment in the elderly. I also review my work related to enhancing visual functions in the normal periphery and how it may be applied to people with central vision loss. PMID- 20010139 TI - Social networking: ethics and etiquette. PMID- 20010142 TI - Hemophagocytic lymphohistiocytosis in the premature neonate. AB - Hemophagocytic lymphohistiocytosis (HLH), a rare disease, results in pathological findings secondary to an abnormal proliferation of activated lymphocytes and histiocytes (tissue macrophages) and is lethal unless identified and adequately treated. Clinical features of HLH include fever, hepatosplenomegaly, cytopenias, hypertriglyceridemia, hypofibrinogenemia, elevated blood levels of ferritin, lymphadenopathy, skin rash, jaundice, and edema. Often, the symptoms of HLH are misinterpreted as infection, resulting in inadequate treatment and death. Several case studies of premature neonates with HLH have recently been published. Therapeutic guidelines for HLH exist and, when identified, HLH in the premature infant can be successfully treated resulting in resolution of symptoms. PMID- 20010143 TI - Hemophagocytic lymphohistiocytosis in the premature neonate: a case study. AB - Hemophagocytic lymphohistiocytosis (HLH) is a rare disease resulting from an abnormal proliferation of histiocytes within the body's tissues leading to an ineffective immune response. Typically, HLH is characterized by fever, hepatosplenomegaly, cytopenia, hypertriglyceridemia, hypofibrinogenemia, and hemophagocytosis. However, the premature infant with HLH may present differently making diagnosis of the disease cumbersome. If an infant is born with ascites, cytopenias, hypofibrinogenemia, and hepatosplenomegaly, a diagnosis of HLH cannot be ruled out. In addition, premature infants oftentimes will not present with fever because they are kept normothermic from ambient sources. Reports of premature infants with HLH in the literature are rare. This is a case presentation of a 27-week-gestation female with a family history of HLH. PMID- 20010144 TI - Recognition and management of the infant with Beckwith-Wiedemann Syndrome. AB - Beckwith-Wiedemann Syndrome (BWS) is the most common overgrowth syndrome in infancy. The characteristic findings are macroglossia, abdominal wall defects, and macrosomia. Genetic studies in infants with BWS demonstrate 3 major subgroups of patients: familial, sporadic, or chromosomally abnormal. Recognition in the neonatal period is important because of the high incidence of childhood malignant tumors associated with BWS. This article provides an overview of the syndrome and discusses its etiology, physical findings, and diagnostic evaluation. Management and clinical implications including family support will also be discussed. PMID- 20010146 TI - Five incredible babies, five paradigm cases that greatly influenced neonatal ethics: what do their parents say today? AB - As the National Association of Neonatal Nurses celebrates its 25th year, we look back at 5 cases that reflect the times in which these babies were born and whose stories have impacted the development of neonatal ethics. These 5 stories take us through the periods of "all must be done" to "parents can choose" and to "are we overdoing?" Neonatal nurses can receive guidance from these wise parents today. PMID- 20010147 TI - Nasogastric tube position and intragastric air collection in a neonatal intensive care population. AB - PURPOSE: For neonates receiving intensive care, nasogastric tube feeding is essential. Since nasogastric tube placement techniques are not well standardized and common verification methods can be unreliable, placement errors may lead to unsafe situations. In mechanically ventilated neonates and neonates on continuous positive airway pressure, malpositioning of the nasogastric tube may prevent excess air within the stomach to escape. In this study, we aimed to relate tube position to amount of air. The hypothesis was: the better the position of the tube, the smaller the amount of air in the stomach. SUBJECTS: A 1-year cohort of neonates in a level IIIc neonatal intensive care unit with a nasogastric tube. DESIGN AND METHODS: We retrospectively reviewed 326 radiographs and classified nasogastric tube position and gastric air. Descriptive statistics were used to describe demographic data. Kendal's tau statistic was applied to explore the relationship between nasogastric tube position and amount of gastric air. A Mann Whitney U test was performed to confirm the differences in gastric air in neonates with Ch5 and Ch6 gastric tubes and neonates with Ch8 gastric tubes. RESULTS: One or both orifices of nasogastric tubes were in the esophagus in 7.1% of cases, tubes were curled up in the stomach in 35.3% of cases, and tube tips were beyond the pyloric sphincter in 5.5% of cases. Substantial or excessive air was found in 37.7% of cases. Kendal's tau value indicated that there was no significant correlation between nasogastric tube position and gastric air. The Mann-Whitney U value indicated that children with Ch5 and Ch6 gastric tubes had significantly more gastric air than children with Ch8 gastric tubes. CONCLUSION: Nasogastric tubes were malpositioned in nearly half of cases, and substantial or excessive air was found in more than one-third of cases. The hypothesis-the better the position of the tube, the smaller the amount of gastric air-was not confirmed by the data. However, a significant relationship was found between tube size and gastric air. PMID- 20010148 TI - Exploring barriers to pain management in newborn intensive care units: a pilot survey of NICU nurses. AB - PURPOSE: To explore barriers that NICU nurses face when attempting to optimally manage newborn pain. SUBJECTS: Ninety California NICU nurses with current membership in the National Association of Neonatal Nurses (NANN) voluntarily participated. DESIGN: A descriptive survey study. METHODS: A researcher-developed survey consisting of 37 questions was mailed to 300 NICU nurses; 102 were returned and 90 were usable. Probability sampling from a listing of California registered nurses with current membership in the NANN was used to obtain the study's sampling frame. PRINCIPAL RESULTS: Less than half of the nurses felt that newborn pain is well managed within the NICUs where they are employed. Barriers identified related to physicians' pain management practices, lack of evidence based pain management protocols, nurses' and physicians' resistance to change practice, infant pain assessment tools, and inadequate staff training regarding pain assessment and management. CONCLUSION: A knowledge-practice gap still exists within newborn pain management. Increased caregiver education remains a necessity, but strategies that address resistance to change practice within healthcare settings must also be considered. PMID- 20010149 TI - NANN Position Statement 3045: Cobedding of twins or higher-order multiples. PMID- 20010150 TI - NANN Position Statement 3046: The use of human milk and breastfeeding in the neonatal intensive care unit. PMID- 20010151 TI - NANN Position Statement 3047: Medication safety in the neonatal intensive care unit. PMID- 20010153 TI - Getting trainees hooked on academic medicine. PMID- 20010154 TI - Interaction between cigarette smoking and high-sensitivity C-reactive protein in the development of coronary vasospasm in patients without hemodynamically significant coronary artery disease. AB - BACKGROUND: Interaction between 2 major risk factors, cigarette smoking and high sensitivity C-reactive protein (hs-CRP), has not been evaluated in patients with coronary vasospasm (CV) without hemodynamically significant coronary artery disease. METHODS: From 1999 to 2005, patients undergoing diagnostic coronary angiography with or without proven CV and without coronary stenosis >50% were evaluated. A total of 621 subjects (335 and 286 with and without CV, respectively) were enrolled in the study. The levels of hs-CRP, measured immediately before coronary angiography, were examined in a subset of 314 patients. RESULTS: Subjects with CV were likely to be older, men, current smokers, and have high hs-CRP levels. The most significant factors for CV were smoking and hs-CRP. In the nonsmoker group, elevated risk of developing CV was only demonstrated in patients with the highest hs-CRP tertile (>5.01 mg/L, P = 0.012). In the smoker group, however, a positively monotonic trend of association was demonstrated between hs-CRP tertile and CV risk, with multivariate-adjusted odds ratios of 1.11, 3.09 (P = 0.012), and 4.12 by the hs-CRP tertiles, suggesting that smokers developed CV at a lower hs-CRP level than nonsmokers and there was a positive interaction between smoking and hs-CRP. CONCLUSIONS: The smokers developed CV at a lower hs-CRP level compared with the nonsmokers. A positive interaction between smoking and hs-CRP was demonstrated for this disease in our study population. PMID- 20010155 TI - The effect of plasma homocysteine levels on clinical outcomes of patients with acute lung injury/acute respiratory distress syndrome. AB - BACKGROUND: Several reports have shown that homocysteine promotes thrombosis by disturbing the procoagulant-anticoagulant balance, whereas alterations in coagulation and fibrinolysis have been suggested as important pathogenetic and prognostic determinants of mortality in acute lung injury (ALI)/acute respiratory distress syndrome (ARDS). The objective of the study was to evaluate the effect of plasma homocysteine levels on the outcomes of patients with ALI/ARDS. METHODS: Sixty-nine consecutive ventilated patients with ALI/ARDS were studied. Blood samples were drawn within 3 days of clinical recognition of ARDS. Measurement of plasma homocysteine, vitamin B12, folate, creatinine, protein C and plasminogen activator inhibitor-1 antigen levels, and genotyping of the methylenetetrahydrofolate reductase gene C677T and A1298C polymorphisms were carried out. The primary outcomes were 28- and 90-day mortality, whereas secondary outcomes included nonpulmonary organ failure-free days, liberation from mechanical ventilation up to day 28, and ventilator-free days during the 28 days after enrollment. RESULTS: In the multivariable analysis, plasma homocysteine concentration adjusted for age, Acute Physiology and Chronic Health Evaluation II score, methylenetetrahydrofolate reductase C677T and A1298C polymorphisms, and levels of plasminogen-activator inhibitor-1 antigen, protein C, creatinine, vitamin B12, and folate was not found to affect significantly mortality at 28 and 90 days (P = 0.39 and P = 0.83, respectively), days without organ failure besides lungs (P = 0.38), the probability of being free from mechanical ventilation at day 28 (P = 0.63), and days without ventilation assistance (P = 0.73). CONCLUSION: Our data suggest that increased plasma homocysteine levels, either alone or in synergy with other thrombophilic risk factors, do not seem to adversely affect the prognosis in patients with ALI/ARDS. PMID- 20010156 TI - C-reactive protein and cardiovascular diseases--is it ready for primetime? AB - C-reactive protein (CRP) is a marker of systemic inflammation, and it has been implicated in the pathogenesis of many chronic diseases, including cardiovascular (CV) diseases. With highly sensitive CRP assays, serum CRP can add considerably to standard coronary heart disease risk factors and in the prediction of subsequent major CV risk. We review evidence supporting the assessment of highly sensitive CRP both in patients with established CV diseases and in those without known disease as well as evidence supporting CRP as a target of therapy. We also review various pharmacologic (especially intensive statin therapy) and nonpharmacologic therapies to reduce levels of CRP. PMID- 20010157 TI - Novel swine-origin (S-OIV) H1N1 influenza A pneumonia in a lung transplant patient: a case report and review of the literature on performance characteristics of rapid screening tests for the S-OIV. AB - Rapid screening tests are insensitive for detecting the novel swine-origin influenza A (H1N1) virus (S-OIV), and false negatives can delay the diagnosis and initiation of appropriate antiviral therapy. The case of a 26-year-old double lung transplant recipient presenting with fever, bilateral pulmonary infiltrates, and a negative influenza direct immunofluorescent antibody on bronchoalveolar lavage is presented. A diagnosis was made, and antiviral therapy was started 10 days after the initial bronchoalveolar lavage on receipt of a positive culture for S-OIV. The published literature on the performance characteristics of rapid screening tests for S-OIV is reviewed in this clinical context. PMID- 20010158 TI - Extensive involvement of the gastrointestinal tract by a de novo presentation of the monoblastic type of myeloid sarcoma: a case report of a rare entity that is often misdiagnosed. AB - We present the first known case of the monoblastic type of myeloid sarcoma (also known as extramedullary myeloid tumor, chloroma, and granulocytic sarcoma) with diffuse involvement of the gastrointestinal tract. The patient originally presented with diarrhea and crampy abdominal discomfort. Endoscopically, the disease showed a diffuse inflammatory process mimicking a number of benign conditions, such as inflammatory bowel disease and autoimmune enteropathy. Sequential biopsies of the upper and lower gastrointestinal tract showed a diffuse infiltrate of increasingly atypical cells. The disease progressed to systemic involvement, including widespread lymphadenopathy, splenomegaly, and pulmonary deposits; the patient died 13 months after the development of initial symptoms. The immunohistochemical and histologic profiles of this case are diagnostic of the monoblastic type of myeloid sarcoma. PMID- 20010159 TI - Long-term complete regression of nodal marginal zone lymphoma transformed into diffuse large B-cell lymphoma with highly active antiretroviral therapy alone in human immunodeficiency virus infection. AB - Since the introduction of combination antiretroviral therapy (cART), there has been a decrease in the incidence of non-Hodgkin lymphoma among the HIV-infected population and also significantly improved survival rates. We describe a remarkable case of a HIV-infected patient whose large B-cell lymphoma, most likely arising by transformation of a nodal marginal zone lymphoma, completely regressed with the use of cART alone. He remained disease-free for almost 3 years and he finally died from presumed flare up of his lymphoma. There are very few cases of spontaneous regression of lymphomas with cART alone in the HIV population. This is an extreme example of the significance of cART in improving survival in HIV-non-Hodgkin lymphoma and changing the face of the HIV epidemic in general. PMID- 20010160 TI - Hyperammonemic encephalopathy: a rare presentation of fibrolamellar hepatocellular carcinoma. AB - Fibrolamellar carcinoma (FLC) is a rare malignant hepatocellular tumor of unknown etiology, arising almost exclusively from noninfected, noncirrhotic liver of young adults. FLC has traditionally been considered to have better survival than hepatocellular carcinoma; however, this notion might be highly erroneous. Patients with metastatic disease at presentation have a dismal prognosis with 5 year survival of only 15%. We describe a case of highly aggressive metastatic FLC that presented as hyperammonemic encephalopathy, which has never been previously reported in the literature. PMID- 20010161 TI - Novel ORC4L gene mutation in B-cell lymphoproliferative disorders. AB - B-cell lymphoproliferative disorders are characterized by marked genetic, morphological, and clinical heterogeneity. The identification of prognostic markers could help to develop risk-adapted treatment strategies. Because proliferation of cells is essential for tumor growth, analysis of the cell cycle might give additional information on tumor progression and clinical behavior. Because initiation of DNA replication represents a significant step in cell division, it is worthwhile to focus the attention to the origin recognition complex (ORC), protein complex essential for initiation of DNA replication. Studies have already shown that ORC-associated factors give a more accurate assessment of cell proliferation than previous markers for many types of malignancies, but so far there have been no studies of eventual role of ORC4L in B-cell lymphoproliferative disorders. Here, we describe 3 patients with B-cell lymphoproliferative disorders (2 with non-Hodgkin lymphoma and 1 with nonsecretory multiple myeloma) carrying a novel A286V mutation within ORC4L gene. All 3 patients were in the advanced stage of disease, but their response to the chemotherapy treatment was good and they achieved complete clinical remission in a relatively short period. Although the functional relevance of this mutation has not yet been elucidated, our observation raises a possibility that A286V mutation, which is constitutively present in these patients, might represent a favorable prognostic marker in B-cell lymphoproliferative disorders. PMID- 20010162 TI - T-cell engineering for cancer immunotherapy. AB - The adoptive transfer of tumor-reactive cells is a promising approach for the treatment of melanoma and some other cancers. To remedy the difficulties associated with the isolation and expansion of tumor-reactive T cells in most cancer patients, peripheral blood T cells can be retargeted to any chosen tumor antigen by the genetic transfer of an antigen-specific receptor. The transduced receptors may be human leukocyte antigen-restricted, heterodimeric T-cell antigen receptor (TCRs), or chimeric antigen receptors (CARs), which typically recognize native cell-surface antigens. Considerable progress has been made in recent years to address the challenges posed by the transfer of either receptor type. Vector and protein modifications enable the expression of TCR chains in human T cells at functional levels and with a reduced risk of mis-pairing with endogenous TCR chains. The combinatorial inclusion of activating and costimulatory domains in CARs has dramatically enhanced the signaling properties of the chimeric receptors described over a decade ago. Based on the effective T-cell transduction and expansion procedures now available to support clinical investigation, improved designer TCRs and second generation CARs targeting an array of antigens are being evaluated in a range of hematological malignancies and solid tumors. PMID- 20010163 TI - From the guest editor: the evolving treatment paradigm in multiple myeloma. PMID- 20010165 TI - The importance of complete response in outcomes in myeloma. AB - Outcomes for patients with multiple myeloma have dramatically improved during the past 20 years as a result of improved therapeutic options and a better understanding of malignant plasma cell biology. Until the past 10 years, the major limitations on improving outcomes were related to the minimal efficacy of existing agents and balancing the toxicity of therapy in an older patient population. However, despite these limitations, there have been advances that have resulted in improvements in progression-free survival and overall survival (OS). High-dose therapy and autologous transplant were the first among therapies to demonstrate an improvement in OS; but more recent analyses have demonstrated that there can be improvement in OS, which is also associated with improvement in the complete response (CR) rate, even among nontransplant patients as well. Thus, achieving CR has been associated with improved OS and has become a therapeutic goal. In the current era of new agents, such as thalidomide, bortezomib, and lenalidomide, the fraction of patients who achieve a CR is now greater than before, and the data regarding the importance of achieving this benchmark of response have never been more benefit. PMID- 20010164 TI - Risk stratification in the era of novel therapies. AB - Multiple myeloma (MM) is an heterogeneous disease and this concept, together with the recent discovery of new drugs with novel mechanisms of action, will lead to the design of individualized treatments. The term "high-risk MM" includes those patients with at least one of the following features: deletion of 17p or t(4;14) or t(14;16), detected by fluorescence in situ hybridization analysis; deletion of 13q detected by conventional cytogenetics; or hypodiploidy or complex karyotype. In addition, patients with high proliferative activity of plasma cells (> or = 3%) measured by the PC labeling index or S-phase by flow cytometry as well as those with a poor response to induction therapy are also high risk. The definition of high-risk MM has been based on patients treated with conventional drugs with or without autologous transplant. However, current data suggest that novel agents can overcome the initial adverse prognosis of deletion 13q and t(4;14) but probably not that of 17p deletion, at least when using immunomodulatory drugs. Nevertheless, the number of patients analyzed is rather limited and, more important, time to progression is only available in a small number of studies. On the basis of these data, it is probably premature to mandate specific therapies on the basis of cytogenetic abnormalities. Moreover, it is possible that the more intensive therapies selected for high-risk patients may be of even greater benefit to standard-risk cases. Accordingly, at present, although we discourage treatment of high-risk patients with conventional schedules, we recommend to include them in large cooperative trials based on novel agents and performing a comprehensive genetic analysis up-front, so that the patients benefiting most from each treatment can subsequently be identified. PMID- 20010167 TI - Integrating novel therapies in the transplant paradigm. AB - The introduction of novel therapies (thalidomide, bortezomib, and lenalidomide) is changing the management of multiple myeloma in younger patients who are usually candidates for autologous stem cell transplantation (ASCT). Randomized trials have shown the superiority of induction treatment with novel agents compared with the classic VAD regimen. Complete remission plus very good partial remission rates of 40% to 60% after induction and of 60% to 75% are achieved with bortezomib-dexamethasone or with 3-drug combinations (including 1 or 2 novel agents). Six randomized studies have evaluated the impact of thalidomide as post ASCT maintenance therapy. Despite differences in the design of the studies and in thalidomide administration, all 6 studies show a significant benefit in the thalidomide arm in terms of response rate and progression-free survival. Results are not as clear in terms of survival. The addition of novel agents before and after ASCT yields very high complete remission and promising progression-free survival rates. On the other hand, results obtained with novel agents are attractive as well and randomized studies comparing novel agents plus early ASCT versus novel agents plus ASCT at progression are planned. PMID- 20010166 TI - Novel therapies in multiple myeloma for newly diagnosed nontransplant candidates. AB - In the twenty-first century, melphalan-prednisone can no longer be regarded as the standard treatment of multiple myeloma patients not eligible for high-dose melphalan followed by autologous stem cell transplantation. The introduction of thalidomide, lenalidomide, and bortezomib has improved the arsenal of therapeutic options in multiple myeloma. Indeed, randomized studies have shown that melphalan prednisone-thalidomide and melphalan-prednisone-bortezomib are superior to melphalan-prednisone alone. In addition, other combinations, including lenalidomide, are under study. In this review, we discuss the role of novel therapies in multiple myeloma in elderly multiple myeloma patients. Important aspects, such as toxicity and the role of prognostic factors, are also addressed. PMID- 20010168 TI - Novel agents for relapsed and/or refractory multiple myeloma. AB - The introduction of novel agents, such as thalidomide, bortezomib, and lenalidomide, has altered the landscape of therapeutic options for multiple myeloma by offering new mechanisms for targeting this disease. Combinations of these agents, with each other and/or traditional chemotherapeutics, have vastly increased the treatment options for patients both frontline, and at relapse, providing higher response rates, and importantly, increasing median overall survival. In this review, we will discuss the use of these novel agents and their combinations in patients with relapsed and/or refractory multiple myeloma. PMID- 20010169 TI - Evolving role of novel agents for maintenance therapy in myeloma. AB - Maintenance therapy after remission achievement is a question still open in multiple myeloma (MM). Steroids and interferon-alpha failed to demonstrate a clear benefit in term of survival. Thalidomide, lenalidomide, and bortezomib have shown to be effective and safe drugs for the treatment of both newly diagnosed and relapsed MM, leading to explore their efficacy also in maintenance setting. Thalidomide seems to be a good choice for patients with low-risk MM and for those who achieved less than very good partial remission after induction treatment. Lenalidomide and bortezomib are still under investigation and a longer follow-up is needed for confirming their role as maintenance treatment. As shown by recent clinical trials, thalidomide and bortezomib are more indicated as consolidation agent, increasing the complete remission/very good partial remission rate. Considering their toxicity profile, first of all peripheral neuropathy and, in case of thalidomide, the lack of correlation between cumulative dose and outcome, a limited administration is suggested. In contrast, lenalidomide showed a low toxicity profile and a benefit from prolonged treatment, making the drug one of the best choices for maintenance treatment. PMID- 20010171 TI - Future novel single agent and combination therapies. AB - Although multiple myeloma (MM) remains an incurable bone marrow cancer, survival rates have dramatically improved over the past decade, most notably in the younger patient population. An understanding of MM biology and improvement in stem-cell transplantation, better supportive care, and novel therapies with higher efficacy and lower toxicity are all responsible for this improvement. Despite these trends, improvements among older patients remain modest, underscoring the need for innovative approaches. The availability of a rich pipeline of novel agents undergoing early-phase clinical trials in MM is an exciting and active area of research. Current novel agents targeting tumor and stromal compartments can be conceptualized as those that target membrane-bound receptors (insulin-like growth factor-1, vascular endothelial growth factor, CD40, etc.), intracellular signaling kinases (Janus kinase/signal transducers and activators of transcription, phosphatidylinositol 3-kinase/protein kinase B/mammalian target of rapamycin, mitogen-activated protein kinase pathways), cell cycle molecular machinery (cyclin-dependent kinases inhibitors), epigenetic abnormalities (DNA methyltransferase and histyone deacetylase), protein dynamics (heat-shock protein 90, ubiquitin-proteasome system), and tumor vasculature and microenvironment (angiogenesis, integrins). This review highlights some of these novel agents tested either alone or in combination for the treatment of MM. PMID- 20010172 TI - Increased expression of vascular endothelial growth factor-C and nuclear CXCR4 in hepatocellular carcinoma is correlated with lymph node metastasis and poor outcome. AB - PURPOSE: Lymph node metastasis (LNM) is a chief cause of morbidity and mortality in patients with hepatocellular carcinoma (HCC) after hepatectomy. The aim of this study was to investigate the relationship between the expression of CXCR4 and vascular endothelial cell growth factor (VEGF)-C and the clinicopathological features of HCC with LNM. METHODS: Immunohistochemical staining for CXCR4 and VEGF-C was performed on tissue microarrays that were constructed using tumor specimens from patients with HCC with (N = 123) or without (N = 145) LNM. The relationship between the clinicopathological features of HCC and the expression of CXCR4 and VEGF-C was analyzed using the Pearson chi(2) test, logistical regression analysis, and receiver operating characteristic analysis. RESULTS: Nuclear CXCR4 expression and VEGF-C expression were positively correlated with LNM and poor outcome in HCC. Moreover, nuclear CXCR4 expression was positively correlated with VEGF-C expression (correlation coefficient 0.256). Receiver operating characteristic analysis revealed that both factors were predictive of HCC LNM {CXCR4: area under the curve, 0.695 [95% confidence interval (CI), 0.630 0.759; VEGF-C: area under the curve, 0.629 (95% CI, 0.562-0.695]}. Patients with tumors exhibiting high nuclear CXCR4 expression or high VEGF-C expression had significantly poorer overall survival than those with low tumor expression of the corresponding factors. Multivariate analysis showed that UICC T stage [odds ratio (OR), 1.615, 95% CI, 1.306-1.997], nuclear CXCR4 expression (OR, 3.998; 95% CI, 2.706-5.907), and VEGF-C expression (OR, 1.903; 95% CI, 1.203-3.011) were independent risk factors for developing HCC LNM. DISCUSSION: These findings suggest that nuclear CXCR4 expression and VEGF-C expression can be used to identify patients with HCC at high risk for developing LNM. PMID- 20010175 TI - Clinical nurse specialists and healthcare reform. PMID- 20010170 TI - Novel immunotherapies. AB - Multiple myeloma is still a fatal disease. Despite advances in high-dose chemotherapy and stem-cell transplantation and the development of novel therapeutics, relapse of the underlying disease remains the primary cause of treatment failure. Strategies for posttransplantation immunomodulation are desirable for eradication of remaining tumor cells. To this end, immunotherapy aimed at inducing myeloma-specific immunity in patients has been explored. Idiotype protein, secreted by myeloma cells, has been the primary target for immunotherapy as it is the best defined tumor-specific antigen. This chapter focuses on novel immunotherapies that are being developed to treat patients with myeloma. I will discuss potential myeloma antigens, antigen-specific T cells, and their function on myeloma tumor cells, and T-cell-based and antibody-based immunotherapies for myeloma. Furthermore, clinical studies of T-cell-based immunotherapy in the form of vaccination, allogeneic stem-cell transplantation and donor lymphocyte infusions, with or without donor vaccination using patient derived idiotype, and future application of donor-derived or patient-derived, antigen-specific T-cell infusion in this disease are also discussed. Based on the specificity of the immune effector molecules and cells, immunotherapies with specific T cells or therapeutic antibodies may represent novel strategies for the treatment of multiple myeloma in the near future. PMID- 20010173 TI - Single-center phase II trial of transarterial chemoembolization with drug-eluting beads for patients with unresectable hepatocellular carcinoma: initial experience in the United States. AB - PURPOSE: This prospective phase II pilot study evaluated safety and efficacy of transarterial chemoembolization (TACE) with drug-eluting beads (DEBs) loaded with doxorubicin in patients with unresectable hepatocellular carcinoma (HCC). METHODS: Twenty patients with unresectable HCC (75% Child's A, 95% Eastern Cooperative Oncology Group performance status 0 to 1, 60% Barcelona Clinic Liver Cancer C, tumor size 6.9 cm) underwent 34 DEB-TACE sessions. Primary endpoints were tumor response, assessed by contrast-enhanced magnetic resonance imaging at 1 month after treatment, using size (response evaluation criteria in solid tumors [RECIST]), contrast-enhancement (European Association for the Study of the Liver) and apparent diffusion coefficient values, and safety assessed by National Cancer Institute Common Terminology Criteria for Adverse Events (NCI CTCAE). Secondary endpoints included feasibility, progression-free survival, and overall survival. RESULTS: DEB-TACE was successfully performed in 34 sessions and demonstrated a favorable safety profile. On initial (1 month) postprocedural magnetic resonance imaging, treated lesions had a mean decrease in size of 4% (P = 0.1129). Using RECIST, partial response was achieved in 2 patients (10%), and 18 patients (90%) had stable disease. Treated tumors demonstrated a mean decrease in contrast enhancement of 64% (P < 0.0001). By European Association for the Study of the Liver criteria, 12 patients (60%) had objective tumor response, and 8 (40%) had stable disease. No patients had progression of a treated lesion while undergoing treatment. At 6 months, the disease control rate was 95% using RECIST. Overall survival rates at 1 and 2 years were 65% and 55%, respectively; median overall survival was 26 months. DISCUSSION: DEB-TACE is safe and effective in achieving local tumor control in patients with unresectable HCC. PMID- 20010176 TI - Using technology to enhance the effectiveness of social networks to promote quality patient care. PMID- 20010177 TI - Lethal outcomes: propofol infusion syndrome and propofol abuse. PMID- 20010178 TI - Building your personal leadership brand. PMID- 20010179 TI - Health promotion in Kenya: a volunteer nurse's experience. AB - This article presents a case study describing how nurses can improve the health behaviors of people living in developing countries. Difficulties and potential solutions are presented. Health promotion allows people to exert control over their health to improve it. A primary difficulty of health promotion in developing countries is communication between care providers and patients. One solution is the utilization of an interpreter; however, in the present study, no professional interpreters were available, thereby complicating the comprehension of new health-related concepts. Another challenge is to understand the patients' perspectives as related to healthcare values. Additionally, as a result of a dearth of evidence-based research in developing countries, difficulties arise in implementing, assessing, and evaluating health promotion programs. Despite these obstacles, nurses continue to travel to developing countries to promote health. Recommendations include respect for a community's health values and incorporation of these values into healthcare planning. To be accepted as a teacher by the local population, the nurse must be able to set aside his/her personal beliefs relating to healthcare, well-being, and disease. Health promotion initiatives should include the means for implementation, thereby enabling the local population to develop skills that will allow them to carry out health promotion projects. PMID- 20010180 TI - The clinical nurse specialist role in developing a geropalliative model of care. AB - PURPOSE: The purpose of this article was to share the experiences and knowledge gained of the clinical nurse specialist's (CNS's) role in the development and implementation of an interdisciplinary geropalliative model of care. BACKGROUND: Across healthcare settings, patients with life-threatening or life-limiting illnesses often experience unwarranted suffering and inattention to their wishes. Studies demonstrate that a palliative approach to care can provide the structure for improved symptom management and earlier identification of patients' goals of care. DESCRIPTION: A palliative model of care was adopted at a 721-bed healthcare facility that encompasses long-term, subacute care, and acute care. The model incorporated a consult team into an embedded approach that provided basic palliative skills in all care. Watson's Caring-Healing Theory guided the model. OUTCOME: Nursing knowledge of palliative care improved, families expressed satisfaction with care, and the staff responded that palliative care positively impacted patient/family outcomes. CONCLUSION: The structure, processes, and outcomes of care can be positively impacted by the CNS during the development and implementation of a palliative model of care. This occurred through CNS activities such as direct consultation, educational initiatives, mentoring, and disseminating assessment and care planning tools. IMPLICATIONS: A geropalliative model of care can be integrated into a variety of healthcare settings. Clinical support and expertise contributed to positive outcomes. Questions for research include the need to identify a symptom assessment tool that is valid, reliable, and easy to use in the chronically ill, geriatric setting, as well as discerning innovative ways to disseminate knowledge to nurses. PMID- 20010181 TI - Clinical nurse specialists as leaders in rapid response. AB - The purposes of this article were to provide background information about rapid response teams (RRTs), to describe the actual and potential outcomes of RRTs, to define the potential role of the clinical nurse specialist in leading RRTs, and to provide recommendations for implementation of RRTs. Rapid response teams provide the opportunity for early intervention for patients demonstrating clinical decline before they reach a point of no return. The long-standing practice of waiting for intensive intervention (code blue) until the patient experiences cardiopulmonary arrest has shown poor outcomes, bringing this practice into question. Although research results are mixed, there is evidence to suggest that the successful use of RRTs results in clinically significant patient outcomes as evidenced by decreased cardiopulmonary arrests and decreased rates in mortality. Other positive patient, nursing, and organizational outcomes can result from RRTs and are discussed. Clinical nurse specialists are uniquely qualified to provide leadership in the development and implementation of RRTs and the monitoring of outcomes. As RRTs become a more common standard practice, further research is needed to examine their benefits and to further refine effective early intervention for high-risk patients. PMID- 20010182 TI - Assessing the health literacy levels of patients using selected hospital services. AB - PURPOSE: The aim of this study was to assess the health literacy abilities of patients in an urban and suburban hospital. BACKGROUND: Inadequate health literacy is a widespread problem that is associated with insufficient self-care knowledge and behavior, inappropriate use of emergency services, higher rates of hospitalization, and increased healthcare costs. Knowledge of patients' reading ability is necessary to ensure that appropriately leveled printed health education materials are available. METHODS: The health literacy of a convenience sample of 21 inpatients and 34 outpatients from an urban and suburban hospital was measured using the Test of Functional Health Literacy in Adults and the Rapid Estimate of Adult Literacy in Medicine. Variables for analysis included demographic characteristics, perceived health status, highest grade of school completed, socioeconomic status, healthcare costs, and number of inpatient and outpatient admissions over the previous year. RESULTS: Based on the Rapid Estimate of Adult Literacy in Medicine, 33% of patients had health literacy levels that were eighth grade or below, whereas according to the Test of Functional Health Literacy in Adults, 23% had marginal or inadequate functional health literacy. Literacy was significantly related to socioeconomic status (P < .001) and education (P < .001), although 30% of participants had a reading level below the highest grade completed. No other significant correlations were found. IMPLICATIONS: Clinical nurse specialists should ensure that health information materials are written at appropriate levels and educate nurses and other healthcare professionals to use effective communication and teaching strategies. PMID- 20010183 TI - Credentialing and psychiatric clinical nurse specialist practice: current issues. PMID- 20010184 TI - Clinical nurse specialist profile. PMID- 20010185 TI - Malignant pertussis in the pediatric intensive care unit. AB - Infantile malignant pertussis is characterized by severe respiratory failure, pulmonary hypertension, leukocytosis, and death. Bordetella pertussis is a vaccine-preventable disease that is becoming a much more globally prevalent condition contributing to more and more infant deaths each year. This article examines 2 such cases and the hospital's internal and community-based programs initiated to help protect these vulnerable infants. PMID- 20010186 TI - Factor V Leiden: a case study and review. AB - Venous thromboembolism accounts for approximately 600,000 hospitalizations and 50,000 deaths per year in the United States. Many inherited blood disorders predispose patients to this disorder. The most common of these disorders is factor V Leiden. Factor V Leiden thrombophilia is an inherited disorder, mutation, of the mechanisms of blood clotting. The mutation causes resistance to activated protein C and thus causes a defect in the natural anticoagulation system. This predisposes patients to recurrent deep vein thrombosis and in combination with other risk factors can cause significant morbidity. This article discusses the pathophysiology, disease characteristics, risk factors for venous thromboembolism, diagnosis and testing, management of factor V Leiden, and implications for nursing in regard to factor V Leiden. The frequency of factor V Leiden and its prevalence in thromboembolic disease emphasize the need for nursing professionals from many diverse backgrounds to better understand the consequences of the factor V Leiden mutation. PMID- 20010187 TI - Candidly critical: real-life critical care nursing, part 1. AB - This is the first in a series of articles depicting real-life situations in critical care nursing. PMID- 20010189 TI - A personal reflection: a case study in family-centered care at the National Naval Medical Center in Bethesda, Maryland. AB - This article describes the family-centered care that the National Naval Medical Center provided to the family members of a war-wounded soldier. It also includes the compelling written thoughts of a mother's bedside vigil during the 32 days her son spent in a sedated and ventilated state in the intensive care unit. PMID- 20010191 TI - Evidence-based practice: fever evaluation and early recognition of systemic inflammatory response syndrome in critical care patients. AB - Fever is very often the first sign of sepsis and systemic inflammatory response syndrome, both of which can be life-threatening. This article presents various guidelines for the treatment and prevention of fever using evidence-based practice guidelines from a variety of sources. PMID- 20010193 TI - Critical care clinical nurse specialist: is this the role for you? AB - This article describes the role of the clinical nurse specialist as an advanced practice nurse in critical care. The characteristics, attributes, roles, and skills of the clinical nurse specialist are presented, as well as an interview with a well-known critical care clinical nurse specialist. PMID- 20010195 TI - Simulating rapid response in undergraduate critical care education. AB - This article describes use of high-fidelity patient simulators to simulate patient emergencies in an undergraduate critical care course. Our rapid response scenarios integrate the nursing process, teamwork, communication, and delegation principles to assist students in recognizing and intervening with patients who are physiologically deteriorating and require urgent nursing care. The scenarios were developed to provide students an understanding of who is involved in rapid response situations, how participants behave, and the roles of the registered nurse and an opportunity to practice those roles. Each senior student participates in 8 to 9 patient scenarios, either in the role of the primary nurse, nursing student, nurse associate, or as an observer during the last semester of their senior year. PMID- 20010197 TI - Implementation of a multidisciplinary ventilator-weaning and sedation protocol in a community intensive care unit. AB - Prolongation of mechanical ventilation poses serious personal and financial threats to healthcare consumers. To that end, many healthcare-related groups have established mechanisms for rapid weaning and subsequent extubation of mechanically ventilated patients. Our objectives were to create and implement an evidenced-based, multidisciplinary care-driven ventilator-weaning protocol as well as revise existing ventilator sedation protocols to decrease length of stay in addition to time spent on the ventilator. Our findings are presented in this article. PMID- 20010203 TI - First impressions. PMID- 20010204 TI - Toward a more disproportionate epidemiology. PMID- 20010205 TI - On the origin of risk relativism. PMID- 20010206 TI - Case-control studies = odds ratios: blame the retrospective model. PMID- 20010207 TI - The hazards of hazard ratios. PMID- 20010208 TI - Epidemiology of Viliuisk encephalomyelitis in Eastern Siberia. AB - BACKGROUND: Viliuisk encephalomyelitis is a disorder that starts, in most cases, as an acute meningoencephalitis. Survivors of the acute phase develop a slowly progressing neurologic syndrome characterized by dementia, dysarthria, and spasticity. An epidemic of this disease has been spreading throughout the Yakut Republic of the Russian Federation. Although clinical, neuropathologic, and epidemiologic data suggest infectious etiology, multiple attempts at pathogen isolation have been unsuccessful. METHODS: Detailed clinical, pathologic, laboratory, and epidemiologic studies have identified 414 patients with definite Viliuisk encephalomyelitis in 15 of 33 administrative regions of the Yakut Republic between 1940 and 1999. All data are documented in a Registry. RESULTS: The average annual Viliuisk encephalomyelitis incidence rate at the height of the epidemic reached 8.8 per 100,000 population and affected predominantly young adults. The initial outbreak occurred in a remote isolated area of the middle reaches of Viliui River; the disease spread to adjacent areas and further in the direction of more densely populated regions. The results suggest that intensified human migration from endemic villages led to the emergence of this disease in new communities. Recent social and demographic changes have presumably contributed to a subsequent decline in disease incidence. CONCLUSIONS: Based on the largest known set of diagnostically verified Viliuisk encephalomyelitis cases, we demonstrate how a previously little-known disease that was endemic in a small indigenous population subsequently reached densely populated areas and produced an epidemic involving hundreds of persons. PMID- 20010209 TI - Biologic risk markers for coronary heart disease: nonlinear associations with income. AB - BACKGROUND: It is unclear whether a linear relationship is an appropriate description of the association between income and biologic markers of coronary heart disease risk. Stronger associations at certain levels of income would have implications for underlying mechanisms. METHODS: The study is based on a healthy sample of 25-64 year olds (n = 14,022) from a nationally representative cross sectional study (the 1988-1994 United States Third National Health and Nutrition Examination Survey). We use regression splines to model the shape of the association between income and 8 biologic markers for coronary heart disease risk, controlling for age, race/ethnicity, marital status, and education. RESULTS: Substantial income-biomarker associations were found for 5 outcomes among women (HDL cholesterol, triglycerides, C-reactive protein, systolic blood pressure, and venous blood lead) and for 3 outcomes among men (HDL cholesterol, triglycerides, and venous blood lead). The most common shapes of association were a stronger association at lower income levels and a greater risk level of biomarker near median income. CONCLUSIONS: We find that the associations of income with biologic risk markers are often nonlinear. The differences in the shape of association suggest there are multiple pathways through which income is associated with coronary heart disease risk. PMID- 20010210 TI - Dioxin exposure and insulin resistance in Taiwanese living near a highly contaminated area. AB - BACKGROUND: Several epidemiologic studies suggest a slightly increased risk of type 2 diabetes in relation to background levels of dioxins. Little is known about how serum dioxins might affect insulin resistance, a hallmark of type 2 diabetes. We examined the association between exposure to dioxins and insulin resistance. METHODS: We investigated 1234 nondiabetic persons living near a deserted pentachlorophenol factory. Using high-resolution gas chromatography/high resolution mass spectrometry and blood biochemistry tests, we measured serum dioxins, fasting glucose, and insulin. Finally, we examined associations between serum dioxin levels and the homoeostasis model assessments of insulin resistance and pancreatic beta-cell function. RESULTS: Participants with insulin resistance (index at or above the 75th percentile) had higher dioxin levels (24.3 vs. 19.8 pg WHO(98)-TEQ(DF)/g lipid) than those without insulin resistance. In both the crude and adjusted models, insulin resistance increased with serum polychlorinated dibenzo-p-dioxins and dibenzofuran (PCDD/F) levels. We found a slight monotonic increase in insulin resistance across the serum PCDD/F categories (P for the trend <0.001). Groups with serum dioxin levels higher than 20.5 pg WHO(98)-TEQ(DF)/g lipid had higher insulin resistance (adjusted odds ratios of 2.7, 3.5, and 5.0 for 50th to <75th, 75th to <90th, and >or=90th percentile, respectively) compared with the reference group (<9.6 pg WHO(98) TEQ(DF)/g lipid [< 10th percentile]). CONCLUSIONS: After adjusting for confounding factors, we found a positive association between serum dioxins and the prevalence of insulin resistance. PMID- 20010211 TI - Vascular brain disease and depression in the elderly. AB - BACKGROUND: Cross-sectional studies have shown an association between vascular brain disease and depression. Longitudinal data are scarce. In a population-based study we investigated this relationship both cross-sectionally and longitudinally. METHODS: Brain MRIs were administered to 479 persons aged 60-90 years at baseline (1995-1996). Brain atrophy, white matter lesions and brain infarcts are all markers of vascular brain disease. At baseline and at follow-up examinations, we also identified persons with depressive symptoms and syndromes using the Center for Epidemiological Studies Depression Scale and psychiatric interviews. Medical records were continuously monitored to identify incident depression. Follow-up was complete until October 2005. RESULTS: At baseline, 36 persons had depressive symptoms. Brain atrophy, white matter lesions, and infarcts were associated with presence of depressive symptoms. During follow-up, 92 persons developed depressive symptoms, 35 of whom were categorized as having depressive syndrome. There was no association of any MRI marker with incident depressive symptoms or syndromes. CONCLUSIONS: Markers of vascular brain disease were associated with depression cross-sectionally. However, when these markers and risk of depression were assessed longitudinally, no relationship was found. PMID- 20010212 TI - Occupational trichloroethylene exposure and kidney cancer: a meta-analysis. AB - BACKGROUND: We conducted a meta-analysis of occupational studies of trichloroethylene-exposed workers to evaluate patterns of associations by study design, exposure assessment methods, and occupational groups. METHODS: Estimates of summary relative risk (RR) were calculated using inverse-variance weighting methods. Cohort studies were classified as group I or group II, depending on quality of the study design and exposure assessment procedures. We conducted sensitivity analyses to examine sources of heterogeneity. RESULTS: Across all studies meeting our inclusion criteria (n = 23), the summary RR was 1.42 (95% confidence interval = 1.17-1.77), with heterogeneity present (test for heterogeneity: P = 0.001). After removal of 3 outlier studies, the summary RR for the remaining studies was 1.24 (1.06-1.45 (test for heterogeneity: P = 0.616)). The summary RR for studies of workers who were identified as more likely exposed to trichloroethylene (group I studies) was 1.34 (1.06-1.68). With outlier studies removed, the group II summary RR estimates for the cohort studies was 0.88 (0.58 1.33) and for the case-control studies was 1.33 (1.02-1.73). The summary RR for studies that used biomarkers to classify exposure (n = 3) was 1.02 (0.59-1.77) and for studies of aerospace/aircraft workers (n = 7) was 1.14 (0.84-1.57). CONCLUSIONS: Positive associations were observed across various study groups. However, considerations of unmeasured potential confounding, lack of quantitative exposure assessment and lack of exposure-response patterns limit epidemiologic insight into the role of trichloroethylene exposure and its potential causal association with kidney cancer. PMID- 20010213 TI - Kidney cancer mortality: fifty-year latency patterns related to arsenic exposure. AB - BACKGROUND: Arsenic in drinking water is associated with kidney cancer. Beginning in 1958, a region of Chile experienced a rapid onset of high arsenic exposure in drinking water, followed by sharp declines when water treatment plants were installed in 1971. METHODS: For the years 1950-1970, we obtained mortality data from death certificates for an exposed region and an unexposed region in Chile. We obtained computerized mortality data for all of Chile for 1971-2000. RESULTS: Kidney cancer risks for the exposed region compared with the unexposed started to increase about 10 years after high arsenic exposures began in 1958. The peak kidney cancer mortality rate ratio (RR) was 3.4 (95% confidence interval = 2.2 5.1) for men in 1981-1985, with subsequent declines to 1.6 (1.2-2.1) by 1996 2000. Mortality RRs among women were 2.9 (1.8-4.7) in 1981-1985 but remained high longer than for men, increasing further to a RR of 4.4 (3.0-6.4) in 1991-1995. Early-life exposure resulted in an increased RR of 7.1 (3.1-14) for young adults aged 30-39 years, born just before or during the high exposure period. CONCLUSIONS: This study shows a latency pattern of increased mortality from kidney cancer, continuing for at least 25 years after the high exposures began to decline. Early life exposure resulted in markedly higher kidney cancer mortality in young adults. PMID- 20010214 TI - Impact of pregnancy-induced hypertension on stillbirth and neonatal mortality. AB - BACKGROUND: Hypertensive disorders of pregnancy are more frequent in primiparous women, but may be more severe in multiparas. We examined trends in stillbirth and neonatal mortality related to pregnancy-induced hypertension (PIH), and explored whether mortality varied by parity and maternal race. METHODS: We carried out a population-based study of 57 million singleton live births and stillbirths (24-46 weeks' gestation) in the United States between 1990 and 2004. We estimated rates and adjusted odds ratios (ORs) of stillbirth and neonatal death in relation to PIH, comparing births in 1990-1991 with 2003-2004. RESULTS: PIH increased from 3.0% in 1990-1991 to 3.8% in 2003-2004. In both periods, PIH was associated with a higher risk of stillbirth and neonatal death. We explored this in more detail in 2003-2004, and observed that the increased risk of PIH-related stillbirth was higher in women having their second or higher-order births (OR = 2.2 [95% confidence interval = 2.1-2.4]) compared with women having their first birth (1.5 [1.4-1.6]). Patterns were similar for neonatal death (1.3 [1.2-1.4] in first and 1.6 [1.5-1.8] in second or higher-order births). Among multiparas, the association between PIH and stillbirth was stronger in black women (2.9 [2.7 3.2]) than white women (2.0 [1.8-2.1]). CONCLUSIONS: A substantial burden of stillbirth and neonatal mortality is associated with PIH, especially among multiparous women, which may be due to more severe PIH, or to a higher burden of underlying disease. PMID- 20010215 TI - Assessing the performance of prediction models: a framework for traditional and novel measures. AB - The performance of prediction models can be assessed using a variety of methods and metrics. Traditional measures for binary and survival outcomes include the Brier score to indicate overall model performance, the concordance (or c) statistic for discriminative ability (or area under the receiver operating characteristic [ROC] curve), and goodness-of-fit statistics for calibration.Several new measures have recently been proposed that can be seen as refinements of discrimination measures, including variants of the c statistic for survival, reclassification tables, net reclassification improvement (NRI), and integrated discrimination improvement (IDI). Moreover, decision-analytic measures have been proposed, including decision curves to plot the net benefit achieved by making decisions based on model predictions.We aimed to define the role of these relatively novel approaches in the evaluation of the performance of prediction models. For illustration, we present a case study of predicting the presence of residual tumor versus benign tissue in patients with testicular cancer (n = 544 for model development, n = 273 for external validation).We suggest that reporting discrimination and calibration will always be important for a prediction model. Decision-analytic measures should be reported if the predictive model is to be used for clinical decisions. Other measures of performance may be warranted in specific applications, such as reclassification metrics to gain insight into the value of adding a novel predictor to an established model. PMID- 20010216 TI - The usefulness of mathematical models in assessing medical tests. PMID- 20010218 TI - Analysis of multiple exposures: an empirical comparison of results from conventional and semi-bayes modeling strategies. AB - BACKGROUND: Analysts of epidemiologic data often contend with the problem of estimating the independent effects of many correlated exposures. General approaches include assessing each exposure separately, adjusting for some subset of other exposures, or assessing all exposures simultaneously in a single model such as semi-Bayes modeling. The optimal strategy remains uncertain, and it is unclear to what extent different reasonable approaches influence findings. We provide an empirical comparison of results from several modeling strategies. METHODS: In an occupational case-control study of lung cancer with 184 exposure substances, we implemented 6 modeling strategies to estimate odds ratios for each exposure-cancer association. These included one-exposure-at-a-time models with various confounder selection criteria (such as a priori selection or a change-in the-estimate criterion) and semi-Bayes models, one version of which integrated information on previous evidence and chemical properties. RESULTS: While distributions of odds ratios were broadly similar across the 6 analytic strategies, there were some differences in point estimates and in substances manifesting statistically significant odds ratios, particularly between strategies with few or no occupational covariates and those with many. Semi-Bayes models produced fewer statistically significant odds ratios than other methods. A simple semi-Bayes model that shrank all the 184 estimates to a common mean yielded nearly identical results to one that integrated considerable prior information. CONCLUSION: Different modeling strategies can lead to different results. Considering the conceptual and pragmatic difficulties of identifying confounders, these results suggest that it would be unwise to place uncritical reliance on any single strategy. PMID- 20010219 TI - In populo. AB - While the Latin phrases in vitro and in vivo are well understood in the medical literature, neither term accurately describes the science performed at the level of the population by epidemiologists and others. In particular, the effect of an exposure in a single organism can differ from those in a population, for reasons ranging from random error to herd immunity. We suggest that in populo (meaning literally "in the people") can fill this gap in the literature. PMID- 20010220 TI - A conversation with Pelayo Correa. Interview by Elizabeth T. H. Fontham. PMID- 20010221 TI - Carlos Juan Finlay: rejected, respected, and right. PMID- 20010222 TI - Am I shrinking? On clothing size and body size. PMID- 20010223 TI - DAG program: identifying minimal sufficient adjustment sets. PMID- 20010224 TI - Validity of self-reported time to pregnancy. PMID- 20010227 TI - Leadership begins within you. PMID- 20010228 TI - Interferon treatment for chronic hepatitis C: a family impact study. AB - Hepatitis C virus is estimated to affect 170 million people worldwide. Infection can lead to cirrhosis, liver failure, or liver cancer. Hepatitis C is unique among chronic illnesses, in that potentially curative treatment is available. Therapy is of prolonged duration and associated with multiple physiological and psychological side effects. These side effects have the potential to impact not only the individual receiving therapy but also their family and the day-to-day functioning of the family unit.This paper describes data and findings obtained from a family impact study instigated to explore the repercussions of interferon treatment for chronic hepatitis C on family life, from both the perspectives of individuals who had received treatment and their family members. An exploratory study was conducted using semi-structured focus groups.Findings reveal the treatment impacted on physical, emotional, relational, and financial domains. The major themes identified were resilience, loss, hardship, anger and irritability, and secrecy. The side-effect profile of therapy exerted significant and previously unforeseen impacts on family relationships, both negatively and positively. Treatment receivers tended to view their experiences as having more adverse impact, while family members, although affected, demonstrated considerable resilience and coping. PMID- 20010229 TI - Characteristics of catheter-related bloodstream infections in children with intestinal failure: implications for clinical management. AB - Catheter-related bloodstream infection is a major cause of mortality and morbidity in the intestinal-failure population. This study reports characteristics of CRBSI with implications for clinical management in parenteral nutrition-dependent children with intestinal failure. The researchers report the rate of central catheter infections, and the causative organisms, as well as identify risk factors in our intestinal-failure patients that would be amenable to preventive measures.The study is a retrospective review of the medical records of 101 patients with intestinal failure (IF), seen in the Intestinal Rehabilitation Clinic at Children's Medical Center of Dallas from May 2005 to March 2007. Catheter-related bloodstream infections (CRBSIs) were categorized as nosocomial or community-acquired. Data collected for each episode include microorganisms isolated from blood and potential risk factors. Z test was done to compare the infection rates.There were 92 episodes of CRBSIs in 45 parenteral nutrition (PN)-dependent patients with central venous catheters (CVC) in place for a total of 13,978 days. Eighty-three percent (n = 76) of CRBSIs developed in the community at a rate of 7.0 per 1,000 days. Seventeen percent (n = 16) nosocomial CRBSIs were observed at a rate of 5.5 per 1,000 catheter days. CRBSI rate was not statistically different between the two groups (7.0 vs. 5.5, p = .378).CRBSI in the intestinal-failure population is due to a wide variety of organisms with numerous risk factors. Education of CVC management with the practice of consistent guidelines may reduce CRBSI incidence, thus reducing the morbidity and mortality in the intestinal-failure patients. PMID- 20010231 TI - Care of older adult populations diagnosed with Helicobacter pylori: a review of current literature. AB - Helicobacter pylori has been linked to development of peptic ulcers and increases the risk of developing gastric carcinoma. Currently, about half of the world's population is infected with H. pylori, many of whom are older adults. Older adult populations often have multiple comorbidities that may mask dyspeptic signs and symptoms. The role of H. pylori and infective signs vary by age, and the confusing symptoms in older adult populations may delay detection and treatment of H. pylori. In addition, time of detection and appropriate treatment is paramount to prevent gastric pathology. Clinicians need to recognize and understand the importance of risk factors, screening, detection, and treatment of H. pylori, particularly in older adults. Earlier detection and more aggressive, proactive clinical recognition of dyspeptic signs and symptoms may lead to earlier screening and diagnosis of H. pylori in older adult populations and help decrease the frequency of gastric metaplasia. PMID- 20010233 TI - The lived experience of treatment for hepatitis C. AB - The purpose of this qualitative study was to explore and describe male patients' experiences of undergoing and completing treatment for the hepatitis C virus. A qualitative, descriptive, and phenomenological methodology was used. Eight male patients diagnosed with the hepatitis C virus, aged 41-60 years and enrolled in a health maintenance organization, were interviewed in their private residences in a community setting. In depth, semistructured, tape-recorded, and transcribed interviews were analyzed by using theme coding. Six themes emerged that described the lived experience of male patients after undergoing treatment for the hepatitis C virus: acquisition of the disease, the diagnosis, the treatment decision making, the "horror stories" regarding treatment, what helped, and feelings now.Patients who are diagnosed with the hepatitis C virus and undergo treatment face a difficult therapy regimen. All participants knew how they acquired the disease and had no sense of apology for how they attracted it. Their only goal was to achieve viral clearance of this disease. Treatment was their option. Nurses play a key role in the management of treatment for patients undergoing therapy for hepatitis C. The management of the adverse effects of therapy has a great impact on patients' quality of life and compliance with the therapy. Nurses are able to help patients by recognizing and managing the adverse effects of this therapy. PMID- 20010234 TI - Endoscopy nurse as a patient care coordinator: the expanded role of the competent nurse in the gastroenterology setting. AB - Several methods are used to describe and evaluate competence profiles of registered nurses. A clinical ladder program based on P. Benner's (1984; P. Benner et al., 1996) competency framework has been used for 20 years in a Finnish university hospital; to ensure the nurses' strategic competencies and capability profiles, the Nurse Competence Scale (NCS) was developed for the self-assessment and the superiors' review purposes (R. Meretoja, H. Isoaho, & H. Leino-Kilpi, 2004; R. Meretoja, H. Leino-Kilpi, & A. M. Kaira, 2004). In the gastroenterological endoscopy unit, competent nurses' work role has been expanded to the proficient level. These nurses are working as patient care coordinators in multiprofessional groups. This expanded work role requires mastery of nursing care for the gastroenterological patients and also familiarity of the endoscopy processes and the clinical pathways. These endoscopy nurse coordinators' most important work role is the effective management of situations, and they are also remarkable resources as preceptors and mentors to less experienced nurses and nursing students. This article describes endoscopy nurse coordinators' expanded work role and competence profile in the frame of the clinical ladder program. PMID- 20010235 TI - Inflammatory bowel disease: patient education using Web-based resources. PMID- 20010236 TI - The tsunami from within: cyclic vomiting syndrome in children. PMID- 20010238 TI - Meperidine usage for endoscopic sedation. PMID- 20010239 TI - SGNA Guideline. Guideline for performance of flexible sigmoidoscopy by registered nurses for the purpose of colorectal cancer screening. PMID- 20010240 TI - Imaging of descemet membrane detachment after trabeculectomy using slit-lamp adapted optical coherence tomography. AB - PURPOSE: To report the use of slit-lamp-adapted optical coherence tomography (SL OCT) in imaging 2 patients with Descemet membrane detachment following anterior chamber reformation after trabeculectomy. METHODS: Two patients, with leaking and overfiltering blebs, respectively, developed Descemet detachments after reformation of flat anterior chambers with viscoelastics. Slit-lamp biomicroscopy and SL-OCT were used to diagnose and monitor posttreatment outcome. RESULTS: Biomicroscopic evaluation and SL-OCT imaging of the cornea revealed a Descemet detachment in both cases. One patient underwent intracameral air injection. Follow-up SL-OCT imaging revealed reattachment of Descemet membrane and reduced corneal thickness. CONCLUSIONS: SL-OCT can be used to document and follow postsurgical detachment of Descemet membrane. PMID- 20010241 TI - Phacoemulsification treatment of subjects with acute primary angle closure and chronic primary angle-closure glaucoma. AB - PURPOSE: To investigate and compare the effects of phacoemulsification as an initial procedure to control intraocular pressure (IOP) in eyes with acute primary angle closure (APAC) and chronic primary angle-closure glaucoma (CPACG) with coexisting cataracts and peripheral anterior synechiae (PAS) of greater than 180 degrees. SETTING: Prospective, cross-sectional study. METHODS: Thirteen patients (13 eyes) with APAC and 10 patients (10 eyes) with CPACG were enrolled in the study. IOP control in the 2 groups was compared at final follow-up. Other outcome measures included visual acuity, anterior chamber depth, gonioscopic appearance, and complications. RESULTS: The postoperative IOP was reduced in both the APAC and CPACG groups. As assessed at last follow-up (median 6 mo), IOP in the APAC group was significantly lower than that in the CPACG group. The absolute success rate was 100% in the APAC group and 80% in the CPACG group. The number of hypotensive medications decreased in both APAC and CPACG group. The postoperative degree of PAS was relieved in 11 of 12 eyes with APAC and 5 of 9 eyes with CPACG. CONCLUSIONS: Initial phacoemulsification performed to provide short-term control of IOP is more efficacious in patients with APAC than in those with CPACG, especially when PAS is greater than 180 degrees. PMID- 20010242 TI - A meta-analysis of topical prostaglandin analogs in the treatment of chronic angle-closure glaucoma. AB - OBJECTIVE: To evaluate the intraocular pressure (IOP)-lowering efficacy of prostaglandin analogs topical medications in patients with chronic angle-closure glaucoma (CACG). METHODS: Pertinent publications were identified through systematic searches of PubMed, EMBASE, KoreaMed, Chinese Biomedicine Database, and the Cochrane-Controlled Trials Register. Randomized clinical trials involving CACG patients treated with latanoprost, bimatoprost, or travoprost monotherapy were selected. Methodologic quality was assessed by a Delphi list with additions and scored out of a maximum of 18. The outcome measures were absolute and relative reduction in IOP from baseline, for diurnal curve, peak, and trough. The pooled effects were calculated using 2-step DerSimonian and Laird estimate method of the random effects model. RESULTS: Nine randomized clinical trials enrolling a total of 1,090 patients were included in the meta-analysis. Quality scores of included studies were generally high, a mean of 14.9 (range, 12-18). The difference in absolute IOP reduction between prostaglandin analogs and timolol varied from 0.4 to 1.6 mm Hg at diurnal curve, 0.9 to 2.3 mm Hg at peak, and 1.3 to 2.4 mm Hg at trough. Relative IOP reduction were diurnal curve, 31% (95% confidence interval, 27% to 34%), peak, 34% (31% to 37%), and trough 31% (28% to 35%) for latanoprost; diurnal curve, 26% (21% to 30%), peak, 28% (24% to 32%), and trough 27% (23% to 30%) for bimatoprost; diurnal curve, 28% (20% to 36%), peak, 32% (31% to 34%), and trough 31% (29% to 33%) for travoprost; and diurnal curve, 23% (19% to 27%), peak, 24% (20% to 29%), and trough 21% (19% to 24%) for timolol. CONCLUSIONS: Latanoprost, travoprost, and bimatoprost provide significant IOP-lowering efficacy in eyes with CACG; and the 3 prostaglandin analogs are associated with at least as effective as timolol. PMID- 20010243 TI - Short-term effect of intravitreal injection of Ranibizumab (Lucentis) on intraocular pressure. AB - PURPOSE: To investigate short-term effect on intraocular pressure (IOP) after intravitreal injection of ranibizumab (Lucentis) (IVIL). MATERIALS AND METHODS: This prospective study included 1 eye of 54 patients (64+/-12 y) with wet age related macular degeneration treated with IVIL. IOP measurements with TonoPen were taken: immediately before and 5 seconds, 5, 10, 15, 30, 60 minutes, and 1 day after IVIL. Axial length (with ultrasound biometry) was assessed in 24 eyes. The analysis included IOP difference at various time points and between phakic and pseudophakic eyes and the relationship between axial length and IOP increases after 5 seconds. RESULTS: Mean IOP were 16.3+/-3.0 mm Hg (range: 12.0 to 21.3), 44.1+/-10.6 (22.0 to 59.3), 29.0+/-9.6 (15.0 to 49.0), 25.8+/-7.9 (16.0 to 45.0), 23.7+/-6.6 (15.7 to 39.0), 21.9+/-5.6 (14.7 to 37.0), 18.8+/-6.1 (10.0 to 35.0), and 16.1+/-3.0 (11.0 to 21.0), respectively, for time points immediately before, 5 seconds, 5, 10, 15, 30, 60 minutes, and 1 day after IVIL. Differences between before IVIL and after IVIL IOP were statistically significant after 5 seconds, 5, 10, 15, and 30 minutes (P=0.0001); however, were not significant after 1 hour (P=0.064) and 1 day (P=0.449). Differences between phakic and pseudophakic eyes were not significant (P>0.05). The relationship between shorter axial length and IOP increase after 5 seconds was significant (linear regression analysis; R=0.28, P=0.007). CONCLUSIONS: IVIL causes a considerable short-term transient rise in IOP. The IOP increase after IVIL can be statistically significant at 0 to 30 minutes after injection in both phakic and pseudophakic eyes, and tends to be greater in shorter eyes. PMID- 20010244 TI - Retinal oxidative stress induced by intraocular hypertension in rats may be ameliorated by brimonidine treatment and N-acetyl cysteine supplementation. AB - PURPOSE: To investigate the effect of brimonidine and N-acetyl cysteine (NAC) on retinal oxidative status under ocular hypertension. MATERIALS AND METHODS: Ocular hypertension is produced in right eyes of 60 rats through intraocular injection of sodium hyaluronate. The left eyes received intracameral saline as sham. Twenty right eyes (brimonidine group) received topical brimonidine twice a day for a week. Other 20 eyes received intraperitoneal NAC (NAC group) once a day. Another group of 20 eyes were followed without any drugs but only intracameral sodium hyaluronate (sodium hyaluronate group) into right eyes. RESULTS: Intraocular injection of sodium hyaluronate increased intraocular pressure for a week and caused retinal peroxidation and decreased glutathione peroxidase and catalase levels. Brimonidine and NAC treatment reversed the retinal oxidative stress created by high intraocular pressure. CONCLUSIONS: Brimonidine and NAC supplementation provide antioxidative properties to retina and decrease retinal damage induced by ocular hypertension. PMID- 20010245 TI - Repeatability and reproducibility for intraocular pressure measurement by dynamic contour, ocular response analyzer, and goldmann applanation tonometry. AB - PURPOSE: To evaluate and compare the intraocular pressure measurement variability between Goldmann applanation tonometry (GAT), Pascal dynamic contour tonometry (DCT), and ocular response analyzer (ORA) tonometry. METHODS: Subjects were prospectively recruited from consecutive Albuquerque VA Medical Center eye clinic patients that were previously diagnosed with ocular hypertension, glaucoma suspect, primary open-angle, or normal pressure glaucoma. Two sets of intraocular pressure measurements (3-4 ORA, 2 DCT, and 2 GAT) were obtained approximately 15 minutes apart. Each set was obtained by 1 of 2 examiners using random examiner sequences. ORA was measured first in both eyes, followed by either DCT or GAT, which were obtained in random order. Repeatability was assessed by examining variability of consecutive measurements by the same examiner, and reproducibility was examined by assessing variability between the first and second measurement sets. RESULTS: One hundred and twenty eyes of 60 subjects were included. Mean age was 64.1+/-9.6 years and 58/60 (97%) were male. Intraobserver repeatability was highest for GAT, followed closely by DCT, and then ORA. Intersession reproducibility was similar for all methods, although a tonographic effect may have corrupted GAT and DCT reproducibility results. We found no repeatability or reproducibility differences between eyes, between examiners, or between measurement sets. CONCLUSIONS: Although some intermethod variability differences were identified, all 3 methods in this study demonstrated clinically acceptable measurement repeatability and reproducibility. This result, in conjunction with the finding that variability was not different between eyes, examiners, or measurement sets, suggests that DCT and ORA are reliable enough to be clinically useful. PMID- 20010246 TI - Outcome of Ahmed valve implantation when preoperative IOP less than 21 mm Hg. AB - PURPOSE: To examine the results of Ahmed glaucoma drainage device (GDD) implantation in patients with a preoperative intraocular pressure (IOP) of 20 mm Hg or less. METHODS: Noncomparative retrospective case series. RESULTS: Sixty-six surgeries in 63 patients were analyzed. Mean follow-up was 51 months. The mean drop in IOP was 3.8 mm Hg (23.3%, P<0.0001) at 12 months and 3.9 mm Hg (24.0%, P<0.0001) at final follow-up. The number of glaucoma medications used postoperatively was significantly less than preoperatively. Although the mean visual acuity was poorer postoperatively this did not reach statistical significance. Surgical success was defined as IOP > or =5 mm Hg and 20% lower than preoperatively with or without hypotensive therapy, visual acuity perception of light or better and no further glaucoma surgery. About 57.6% and 53.0% of patients were considered a success at 12 months and final follow-up, respectively. Fourteen patients (21.2%) underwent additional glaucoma surgery, 8 of which had a second Ahmed GDD inserted whereas 6 had cyclodestruction. All failures were due to inadequate IOP control or further glaucoma surgery, with no cases considered failures due to vision loss or hypotony. Perioperative complications developed in 6 cases (9.1%) whereas long-term problems related to surgery occurred in 8 cases (12.1%). CONCLUSIONS: In patients with IOPs of 20 mm Hg or less who require IOPs in the low teens and below Ahmed GDD surgery does seem to be an effective option. The success rates of surgery, however, must be balanced against the risk of complications. In addition, the relative merits of Ahmed GDD implantation versus mitomycin trabeculectomy and/or nonvalved GDDs requires further investigation. PMID- 20010247 TI - Trabeculectomy in patients with primary angle-closure glaucoma. AB - PURPOSE: To analyze the results of trabeculectomy in patients with primary angle closure glaucoma. METHODS: A retrospective and noncomparative case series analysis was performed on data from Taiwan, from 2001 to 2004. The outcomes of trabeculectomy in eyes with acute primary angle-closure glaucoma attack (AACG) and those with chronic primary angle-closure glaucoma (CACG) were assessed in terms of final intraocular pressure (IOP), changes to visual acuity, and the incidence of complications. RESULTS: A total of 52 eyes of 52 patients, 15 patients in AACG group and 37 patients in CACG group, were reviewed. The mean follow-up period was 32 months (range, 26-42 mo). In terms of no visual acuity change after surgery, there were significant differences between the CACG and AACG groups (P=0.02, Fisher exact test). In terms of final IOP control, trabeculectomy outcome was significantly worse in patients in the AACG group than those in the CACG group (P<0.01, Fisher exact test). Moreover, the complication of bleb encapsulation appeared more frequently in the AACG group than in the CACG group (P=0.02, Fisher exact test). CONCLUSIONS: Compared with CACG, trabeculectomy may not be as good for patients of AACG, because it may lead to worsened visual acuity; it also seems to present a greater failure rate, more complications, and fewer cases of surgical survival. PMID- 20010248 TI - Normotensive glaucoma and risk factors in normotensive eyes with pseudoexfoliation syndrome. AB - PURPOSE: To evaluate the prevalence of normotensive glaucoma (NTG) in patients with pseudoexfoliation (PEX) syndrome. METHODS: In this prospective study, 178 eyes of 178 patients with PEX syndrome who had no previous history of antiglaucomatous treatment were evaluated in terms of NTG prevalence. NTG diagnosis was based on the presence of glaucomatous optic nerve head damage with glaucomatous visual field defects, open angles, and intraocular pressure (IOP) levels less than 22 mm Hg. Two groups of normotensive pseudoexfoliative eyes with or without glaucomatous findings were compared in terms of age, sex, the presence of systemic diseases and mean IOP, maximum IOP, and IOP fluctuation levels. RESULTS: The IOP levels were below 22 mm Hg in 109 of the 178 eyes, and 24% (n=27) of those eyes were diagnosed as NTG. There were no significant differences between the eyes with or without glaucoma in terms of age, sex, and the presence of systemic diseases (P>0.05). The mean IOP, maximum IOP, and IOP fluctuation levels were significantly higher in eyes with NTG when compared with eyes without glaucoma (P<0.05). Univariate analysis revealed that significant risk factors associated with NTG in patients with PEX syndrome were higher mean IOP (> or =14 mm Hg), higher maximum IOP (> or =18 mm Hg), and greater IOP fluctuation (> or =4 mm Hg). In multivariate analysis, these factors were higher maximum IOP and greater IOP fluctuation range (P=0.003, P<0.001, respectively). CONCLUSIONS: There are glaucomatous findings in a significant proportion of normotensive patients with PEX syndrome. The high maximum IOP levels and greater IOP fluctuation range may be contributory factors in the pathogenesis of NTG in patients with PEX syndrome. PMID- 20010249 TI - Effect of trabeculectomy on intraocular pressure fluctuation with postural change in eyes with open-angle glaucoma. AB - PURPOSE: To investigate the effect of trabeculectomy on the intraocular pressure (IOP) fluctuations caused by the postural change in patients with open-angle glaucoma. METHODS: Prospective, nonrandomized study. Ten eyes of 10 patients with primary open-angle glaucoma and 3 eyes of 3 patients with normal-tension glaucoma who underwent trabeculectomy were studied. The IOP at the baseline and 3 months after trabeculectomy was measured in both sitting and supine positions. A pneumatonometer was used to measure the IOP in both sitting and supine position. RESULTS: The mean IOP in the sitting position was 19.6+/-5.1 mm Hg at the baseline and 9.4+/-2.9 mm Hg after trabeculectomy (P<0.01, paired t test). The mean difference in IOP between the sitting and supine position was 4.1+/-1.6 mm Hg at the baseline and 2.2+/-1.5 mm Hg after trabeculectomy (P=0.01, paired t test). CONCLUSIONS: Trabeculectomy might decrease not only the IOP in the sitting position but also the magnitude of IOP elevation associated with the postural change. PMID- 20010250 TI - The promise and future of comparative effectiveness research. PMID- 20010252 TI - Communicating quality via "road shows". PMID- 20010251 TI - Improving care in nursing homes using quality measures/indicators and complexity science. PMID- 20010253 TI - Aspirin for the prevention of cardiovascular disease: systematic review. PMID- 20010254 TI - Keeping the NCLEX-RN Current. PMID- 20010255 TI - Speaking with the press. AB - Many new nursing leaders assuming deanships or assistant or interim deanships have limited education, experience, or background to prepare them for the job. To assist new deans and those aspiring to be deans, the authors of this department, 2 deans, offer survival tips based on their personal experiences and insights. They address common issues, challenges, and opportunities that face academic executive teams, such as negotiating an executive contract, obtaining faculty lines, building effective work teams, managing difficult employees, and creating nimble organizational structure to respond to changing consumer, healthcare delivery, and community needs. The authors welcome counterpoint discussions with readers. PMID- 20010256 TI - Success begins in nursing freshman orientation course. PMID- 20010257 TI - Assisting your students to work with Spanish-speaking clients. PMID- 20010258 TI - Baby books: a tool for teaching and learning. PMID- 20010259 TI - Is tax on soft drinks in our future? PMID- 20010260 TI - Assisting nursing students to develop empathy using a writing assignment. PMID- 20010261 TI - Are you teaching your students strategies related to bullying in the nursing workforce? PMID- 20010262 TI - Assessing the quality of multiple-choice test items. AB - With the focus of nursing education geared toward teaching students to think critically, faculty need to assure that test items require students to use a high level of cognitive processing. To evaluate their examinations, the authors assessed multiple-choice test items on final nursing examinations. The assessment included determining cognitive learning levels and frequency of items among 3 adult health courses, comparing difficulty values with cognitive learning levels, and examining discrimination values and the relationship to distracter performance. PMID- 20010263 TI - Using data from the internet to teach ethical principles for critiquing research studies. AB - Qualitative researchers are increasingly using the Internet to conduct their studies; however, they need to adhere to the same ethical principles used when conducting traditional research studies. Students also must be aware of these ethical principles and know how to critique research reports for adherence to them. The authors describe selected ethical principles as they relate to Internet qualitative research, what students need to know, what faculty must teach them to critique those studies, and what decisions students must make once the critique is completed. PMID- 20010264 TI - Evidence-based curricular strategies to enhance BSN graduates' transition into practice. AB - A qualitative analysis of 1-year alumni surveys provided the basis for developing evidence-based curricular strategies to better prepare BSN graduates for their transition into practice. The authors discuss study results, curricular strategies initiated, and beginning feedback regarding their effectiveness in preparing BSN graduates for practice. PMID- 20010265 TI - Are students as good at multitasking as they think? PMID- 20010266 TI - Teaching undergraduate nurses nursing phenomena. AB - Nursing phenomena, complex and dynamic conceptual building blocks, are the basis of our nursing language. Students have difficulty understanding what nursing phenomena are and how to link them to practice and research. The authors describe incremental experiential learning strategies that were used to help students learn and apply the basic concepts of their nursing language. PMID- 20010267 TI - $33 million available to train health professionals. PMID- 20010268 TI - Journal editor survey: information to help authors publish. AB - Faculty are expected to publish but often do not know how to write a manuscript that avoids major reasons for subsequent rejection. The authors share the results of a survey of 63 journal editors who reported journal characteristics and reasons for rejection of manuscripts. Suggestions to improve the acceptance rate of faculty publications are offered. PMID- 20010269 TI - Is that itch really a pain? PMID- 20010270 TI - Clinical preceptors enhance an online accelerated bachelor's degree to BSN program. AB - The accelerated online BSN program immerses students into the real world of nursing by presenting all theory courses online and almost all clinical experiences using a preceptor model. A rigorous "boot camp" jump starts the student's clinical learning experience and is followed by a variety of precepted experiences. The authors discuss the collaborative roles, distinctive program characteristics, and overwhelmingly positive outcomes. Graduates have high NCLEX scores and low attrition rate and acclimate easily into their nursing professional role. PMID- 20010271 TI - Using a writing group to promote faculty scholarship. AB - Writing productivity is an essential component of scholarship. Barriers to writing include intrapersonal characteristics, faculty role complexity, and time constraints. Writing groups can increase faculty members' writing, contributing to dissemination of nursing knowledge and advancement of professional nursing. The authors discuss the structure and processes of a writing group that can be adapted by faculty interested in using comentoring to increase their scholarship. PMID- 20010272 TI - Evidence-based practice in the military healthcare system. PMID- 20010274 TI - Evidence-based practice: a civilian perspective. PMID- 20010273 TI - Collaborating across services to advance evidence-based nursing practice. AB - BACKGROUND: Military medical treatment facilities offer a unique environment in which to develop a culture of evidence-based practice (EBP). Distinctive issues arise in the context of changed patient care demographics because of a war injured population. These issues offer an opportunity to enhance the quality of care through the use and adaptation of research findings in this special nursing environment. In addition, the colocation of two military medical centers offers the prospect of collaborative efforts to create a regional culture for nursing EBP. OBJECTIVES: The purposes of this study were to describe the processes of a collaborative project to train nurses in EBP and to share resources in developing and implementing evidence-based clinical nursing guidelines in two large military medical centers in the Northeastern United States and to discuss the collective efforts of nurse researchers, leadership, advanced practice nurses, and staff nurses in each hospital to facilitate the EBP process. METHODS: A description of the organizational structure and the climate for EBP of each facility is provided followed by discussion of training efforts and the inculcation of an organizational culture for EBP. RESULTS: Contextual barriers and facilitators were encountered throughout the project. The two nurse researchers leading the projects were able to overcome the barriers and capitalize on opportunities to promote EBP. Three evidence-based clinical practice guidelines were developed at each facility and are currently in various stages of implementation. DISCUSSION: Despite the barriers, EBP continues to be at the forefront of military nursing practice in the U.S. National Capital Region. Clear communication and regular meetings were essential to the success of the collaborative project within and between the two military hospitals. Military-specific barriers to EBP included high team attrition and turnover because of the war mission and the usual high staff turnover at military hospitals. Military facilitators included a common mission of providing high-quality care for war-injured service members. Lessons learned from this project can be generalized to civilian facilities. PMID- 20010275 TI - Care of the patient with enteral tube feeding: an evidence-based practice protocol. AB - BACKGROUND: Care of patients with enteral feeding tubes often is based on tradition and textbook guidance rather than best evidence. Care practices can vary widely both between and within institutions, and this was the case at a northeastern military medical center that served as the site for this evidence based protocol development and implementation project. OBJECTIVES: The purpose of this study was to describe the development and implementation of an evidence based clinical protocol for care of patients with enteral feeding tubes. METHODS: This was an evidence-based implementation project with pretest-posttest measures. Protocol data collection occurred both before and after implementation of the protocol. Data collection tools were based on the literature review and included three domains: (a) documentation of patient procedures, (b) nursing knowledge of each of the specific procedures, and (c) environment of care. Descriptive statistics and data were analyzed using independent samples t tests. RESULTS: Overall staff knowledge of enteral feedings and methods used to unclog both large and small-bore feeding tubes differed significantly before and after implementation (p < .05). Staff knowledge regarding the danger of using blue dye in feeding solution was significant (p < .001). There was improvement also in administration of medications separately rather than mixed together and in head of bed elevation of patients with feeding tubes. There was a 10% improvement in documentation of patient family education and a 15% improvement in recording fluid flushes during medication administration. After implementation, environment of care data collection showed 100% of patients with head of bed elevated and with functioning suction available, an improvement over levels before implementation. DISCUSSION: Care must be taken in the interpretation of these findings because it was generally not the same nurses who answered both surveys. High staff turnover within this military hospital also affected sustainment of the protocol implementation. Maintenance activities must be constant and visible within the organization. A champion for evidence-based practice greatly enhances uptake and maintenance of nursing practice change. PMID- 20010276 TI - Evidence-based practice in a military intensive care unit family visitation. AB - BACKGROUND: The dissatisfaction of family members with a restrictive visiting policy in a combined intensive care and step-down unit provides an opportunity for staff to develop better ways to meet the needs of patients and their families. A review of the evidence-based practice (EBP) literature as the measure of significance indicated that less restrictive visitation policies enhance patient and family satisfaction and offer many physiologic and psychological benefits to the patient. OBJECTIVE: The purpose of this study was to determine whether a less restrictive visitation policy could be implemented in the adult critical care department of a naval medical center. METHODS: Staff was educated on the use of EBP, specifically the Iowa-based model, and a less restrictive visitation policy was developed and implemented. RESULTS: Evaluation of the postintervention survey findings revealed higher patient and family satisfaction. CONCLUSIONS: The findings from this EBP project suggest that a more open visitation policy is feasible in adult critical care units, with an increase in overall satisfaction of patients and their families with regard to their stay in the intensive care unit. PMID- 20010277 TI - Implementing a unit-level intervention to reduce the probability of ventilator associated pneumonia. AB - BACKGROUND: Ventilator-associated pneumonia (VAP) is the second most common hospital-acquired infection and is associated with high morbidity and mortality rates for mechanically ventilated patients in the intensive care unit. Routine nursing interventions have been shown to reduce VAP rates. OBJECTIVES: The purpose of this study was to evaluate the effectiveness of a unit-specific education intervention that emphasized hand hygiene, head-of-the-bed elevation, and oral care. The goals were to improve staff compliance with hand washing, head of-the-bed elevation, and oral care; to decrease VAP rates, and to decrease number of ventilator days. METHODS: Two-hour observations were conducted on a convenience sample of 100 ventilated patients not diagnosed with VAP and the clinical staff that interacted with them. Instrumentation included a compliance checklist, a demographic patient survey, and the Acute Physiology and Chronic Health Evaluation AEIV tool. Unit-specific educational interventions were designed and implemented on each participating unit. RESULTS: : The VAP and the ventilator day rates did not improve significantly. There were no significant changes in clinician adherence to hand hygiene, provision of oral care, or patient positioning. DISCUSSION: Despite implementation of both structured and creative education, team-based approach, and frequent staff reminders, patient outcomes and staff compliance did not improve significantly. Unit-based education interventions may not be the best strategy to facilitate change. Organizations with frequent changes in personnel and leadership may not have the unit-level infrastructure necessary to attain and sustain change. PMID- 20010278 TI - "Stars" aligned for evidence-based practice: a Triservice initiative in the Pacific. AB - BACKGROUND: Establishing system-wide evidence-based nursing practice requires the support of senior leadership as well as adherence to a model that is sensitive to the challenges of practice change. OBJECTIVE: The purpose of this study was to establish a level of skill and knowledge among Army, Navy, and Air Force nurses to develop, to implement, and to evaluate best evidence-based practices (EBPs) for improved patient care. METHODS: Using the Iowa Model to guide their work, nurses from the Air Force, the Army, and the Navy joined efforts to establish EBP as a standard of nursing practice at four different military healthcare facilities in Hawaii: one tertiary medical center and three clinics on the island of Oahu. With the support of senior leadership at all four sites, four nurse researchers and a project director facilitated the work of advance practice nurses and staff nurses. RESULTS: Twelve different EBP projects were developed, implemented, and evaluated over 18 months by more than 50 nursing personnel across the three branches of service on Oahu. The result was a positive contribution to patient care throughout the healthcare system, affecting all areas of military nursing practice. Supporting goal was the improvement in the attitudes of nurses about research and their skills in applying evidence to their practice by way of a year-long internship program. Also influenced was the development of a foundation for evidence-based clinical and administrative military nursing decision making in the Pacific. CONCLUSION: A successful EBP program requires (a) consistent long-term vision and support from senior leadership; (b) resources such as trained personnel, finances, and time; and (c) structured approach to create an environment that facilitates the process. However, the success of an EBP program relies heavily on each nurse's commitment to change designed to improve nursing care for patients. PMID- 20010279 TI - A nurse-facilitated depression screening program in an Army primary care clinic: an evidence-based project. AB - BACKGROUND: Depression, sometimes with suicidal manifestations, is a medical condition commonly seen in primary care clinics. Routine screening for depression and suicidal ideation is recommended of all adult patients in the primary care setting because it offers depressed patients a greater chance of recovery and response to treatment, yet such screening often is overlooked or omitted. OBJECTIVE: The purpose of this study was to develop, to implement, and to test the efficacy of a systematic depression screening process to increase the identification of depression in family members of active duty soldiers older than 18 years at a military family practice clinic located on an Army infantry post in the Pacific. METHODS: The Iowa Model of Evidence-Based Practice to Promote Quality Care was used to develop a practice guideline incorporating a decision algorithm for nurses to screen for depression. A pilot project to institute this change in practice was conducted, and outcomes were measured. RESULTS: Before implementation, approximately 100 patients were diagnosed with depression in each of the 3 months preceding the practice change. Approximately 130 patients a month were assigned a 311.0 Code 3 months after the practice change, and 140 patients per month received screenings and were assigned the correct International Classification of Diseases, Ninth Revision Code 311.0 at 1 year. The improved screening and coding for depression and suicidality added approximately 3 minutes to the patient screening process. The education of staff in the process of screening for depression and correct coding coupled with monitoring and staff feedback improved compliance with the identification and the documentation of patients with depression. Nurses were more likely than primary care providers to agree strongly that screening for depression enhances quality of care. DISCUSSION: Data gathered during this project support the integration of military and civilian nurse-facilitated screening for depression in the military primary care setting. The decision algorithm should be adapted and tested in other primary care environments. PMID- 20010280 TI - Improving compliance with diabetes clinical practice guidelines in military medical treatment facilities. AB - BACKGROUND: A multidisciplinary, multifaceted approach to disease management that incorporates the health system, the provider, and the patient is supported in the literature. There was a need to improve patient outcomes to meet or to exceed the Health Plan Employer Data and Information Set (HEDIS) benchmarks for the management of patients with diabetes. OBJECTIVES: The purpose of this study was to implement a process improvement effort using practice guidelines on the basis of an evidence-based practice model for the management of type II diabetes mellitus at two primary care clinics at two military medical facilities in Hawaii. METHODS: A retrospective review of charts, electronic records, and system data revealed that the clinics used as project sites were not compliant with established guidelines for diabetes management. After a literature review and an analysis of the current processes, a multidisciplinary care delivery model was developed and implemented to identify spheres of influence involving all members of the diabetes management team and the tasks that influenced patient outcomes. RESULTS: Improvements were seen for more than 6 months of initial practice change, including compliance with annual glycosylated hemoglobin (HbA1c), lipid, blood pressure, and foot checks. At Site 1, HEDIS measures increased for adequately controlled HbA1c and low-density lipoprotein (LDL) from 80% to 85% and from 49% to 58%, respectively. Site 2 showed an increase in adequate control of HbA1c from 77% to 79% at 6 months. After a steady increase in compliance, the percentage for adequately controlled LDL dropped to 56% at 9 months. At Site 1, HEDIS measures decreased slightly to 82% for HbA1c control and to 54% for LDL control at the 9-month mark. DISCUSSION: Inconsistent delivery of care and lack of staff and patient involvement influenced process outcomes. There were challenges with database accuracy, adequate staffing, computer software upgrades, and overseas site locations. Annual foot examinations showed the largest improvement over time. Site 1 had a significant increase in filament testing because of an innovative strategy to develop a competency program to educate technicians to perform the assessment during the patient check-in process. Sustainability is needed to improve overall patient quality and patient safety and to decrease variation in care among medical treatment facilities over time. PMID- 20010281 TI - The evidence-based practice concept: engaging interest and participation. AB - The TriService Nursing Research Program was chartered in 1992 to advance the science of military nursing to support mission readiness, to provide optimal evidence-based healthcare in various military settings, and to improve the health and quality of life of military personnel. In 2002, the Tri-Service Nursing Research Program created the evidence-based practice initiative to foster excellence in military nursing care through the evidence-based practice in the military healthcare system. The evidence-based practice initiative has been coherently and successfully implemented throughout the military healthcare systems largely due to strong commitment and support from all management levels across services and across components. PMID- 20010282 TI - Facilitation of evidence-based nursing practice during military operations. AB - The translation of research to clinical practice and health decision making is challenging. Under military operational conditions (e.g., the provision of care in the field), translation may be even more challenging. Two barriers that limit the use of evidence to guide practice, which are particularly germane under operational conditions, are conflicting or absent research results specific to the population of interest and relevant studies not being compiled in one place (Titler, 2007; Titler & Everett, 2001). The Agency for Healthcare Research and Quality Knowledge Transfer Framework (Nieva et al.,2005) provides a structure to facilitate evidence translation and to overcome these barriers. This article summarizes one aspect of a program of operational nursing research supported by the TriService Nursing Research Program, which exemplifies the three stages of the Agency for Healthcare Research and Quality framework. PMID- 20010283 TI - Mantle cell lymphoma with skin invasion characterized by the common variant in the subcutis and blastoid transformation in the overlying dermis. AB - We report a case of common mantle cell lymphoma (MCL) with subcutis infiltration and transformation to blastoid MCL in the overlying dermis. The patient was initially diagnosed as having chronic lymphocytic leukemia and treated with chemotherapy. Eight months after the diagnosis of MCL with bone marrow involvement, subcutaneous nodules developed on the patient's left thigh and forearm. A skin biopsy showed a massive infiltration of neoplastic lymphocytes throughout the dermis and subcutaneous tissue. In the upper dermis, there was a perivascular mixed infiltrate of atypical large lymphoid cells and small-sized cells. In the mid to lower dermis, the infiltrate was dense with a nodular growth pattern and was composed of atypical large lymphoblast-like cells with large nuclei, dispersed chromatin, and numerous mitoses. In the subcutaneous tissue, there was a diffuse infiltration of neoplastic cells with common MCL cytologic features characterized by small- to medium-sized lymphoid cells. Cells in the common and blastoid variants of MCL were immunohistochemically positive for CD20 and cyclin D1 but negative for CD5. Neoplastic lymphocytes from the patient's bone marrow had the typical morphologic features and the immunophenotype of MCL (ie, CD5, CD20, cyclin D1, CD10, and CD23). Other case reports in the medical literature indicate that an MCL with skin invasion tends to have a poor prognosis. Our patient died 3 months after the appearance of skin invasion. PMID- 20010284 TI - Dermatopathic lymphadenitis with generalized erythroderma in a patient with epstein-barr virus-associated hemophagocytic lymphohistiocytosis. AB - Here, we describe a patient with Epstein-Barr virus (EBV)-associated hemophagocytic lymphohistiocytosis (HLH) who simultaneously presented with generalized erythroderma and dermatopathic lymphadenitis (DL). A 63-year-old Korean woman presented at our hospital with fever, hepatosplenomegaly, axillary lymphadenopathy, and generalized erythrodermic eruption. The bone marrow biopsy findings were consistent with the diagnosis of HLH, and EBV DNA was detected using the polymerase chain reaction. Based on serologic tests that indicated a primary EBV infection, the patient was diagnosed with EBV-associated HLH. Histopathologic analysis of enlarged lymph nodes was consistent with DL, and EBV encoded small nuclear RNA-positive mononuclear cells were detected. We assume that activated histiocytes, lymphocytes, and proinflammatory cytokines in HLH may have important roles in the development of generalized erythroderma and DL. Disrupted epidermal/dermal junctions owing to erythroderma may also be involved in the development of DL. PMID- 20010285 TI - H syndrome: recently defined genodermatosis with distinct histologic features. A morphological, histochemical, immunohistochemical, and ultrastructural study of 10 cases. AB - This study analyzes the histopathological findings in H syndrome, a recently recognized autosomal recessive genodermatosis characterized by indurated, hyperpigmented, and hypertrichotic skin in well-defined anatomical areas accompanied by various systemic manifestations. So far, descriptions of the histopathological skin changes in this disorder, as reported in a few small case series, were inconsistent, leading to diverse clinical interpretations. In an attempt to define standardized, diagnostic, morphological criteria that will distinguish this disorder from other fibrosing conditions, we studied skin biopsies from 10 patients with H syndrome. The characteristic morphology included widespread fibrosis (moderate in dermis and severe in subcutis); striking mononuclear infiltrates consisting mainly of monocyte-derived cells (small CD68 histiocytes and CD34 and FXIIIa dendrocytes) and plasma cells; and thickened, fragmented, and partially calcified elastic fibers, admixed with well-formed psammoma bodies, a previously unrecognized feature in nonneoplastic skin and subcutaneous conditions. In addition, the ultrastructure of CD68 small histiocytes exhibited distended endoplasmic reticulum and scarcity of lysosomes, features typical for fibroblasts but unusual for histiocytes. These unusual findings in the histiocytes pose a question as to their possible role in the fibrotic cascade in this disorder. We conclude that the above findings are essential for the diagnosis of H syndrome and that incisional biopsies are mandatory for recognition of the full spectrum of histopathological findings. PMID- 20010287 TI - Sudden death from massive gastrointestinal hemorrhage associated with crack cocaine use: case report and review of the literature. AB - Cocaine is associated with many health complications including gastrointestinal ischemia/infarction, and hemorrhage. The clinical medical literature describes such entities, but no case exists within the forensic literature describing life threatening intraluminal gastrointestinal hemorrhage and sudden death associated with cocaine. A 48-year-old black man died suddenly in his home, with drug paraphernalia near the body. Further investigation revealed blood stains on upholstery, and a toilet filled with fresh blood. External examination of the body revealed fresh blood draining from the anus. Internal examination revealed a dusky portion of distal ileum filled with an abundance of fresh blood. Toxicology confirmed the suspicion of acute cocaine intoxication.Review of the clinical literature revealed several case reports describing an association between cocaine use and intestinal infarction, one with hemoperitoneum. Review of the forensic literature returned one case describing an association between cocaine toxicity and intestinal infarction, but without hemorrhage. To our knowledge, our case represents the first report in the forensic literature describing massive gastrointestinal hemorrhage associated with acute cocaine intoxication. It is important that the forensic pathologist recognize cocaine toxicity as a potential cause of gastrointestinal bleeding, especially in younger male decedents without significant comorbidities. Scene investigation may reveal useful clues pointing to this unusual etiology. PMID- 20010286 TI - A fatal case of a paint thinner ingestion: comparison between toxicological and histological findings. AB - Toluene and xylene are aromatic hydrocarbons commonly used as an industrial solvent for the manufacturing of pharmaceuticals, paints, and chemicals. The Occupational Safety and Health Administration has determined that toluene levels of 2000 parts per million (ppm) are considered dangerous to life and health. Several studies have examined the absorption of toluene and xylene following inhalation and oral ingestion in humans. Volatile organic compounds that are absorbed into the blood are distributed throughout the body; in particular, distribution of absorbed toluene and xylene in humans and rodents is characterized by preferential uptake in well-perfused and lipophil tissues such as the brain, liver, lungs, and body fat and also in central nervous system. The available studies indicate that xylenes are rapidly absorbed independently from the kind of exposition. We illustrate a fatal case of self-poisoning by ingestion of varnishes diluting solvents, reporting the identification and quantification of volatile organic compounds (toluene, o-m-p xylene) from human biologic liquids and viscera samples using the Solid-Phase Microextraction-Headspace-Gas Chromatography/Mass Spectrometry to toxicological analysis, and the histopathological findings evaluated in liver, kidney, and lungs. PMID- 20010288 TI - Sudden death caused by fibrovascular esophageal polyp: case report and study review. AB - Benign tumors of the esophagus are rare but can lead be fatal when aspirated into the upper airway. Here, we describe a sudden death due to aspiration of a fibrovascular polyp supported by a review of 11 cases from the literature. PMID- 20010289 TI - Acute hemorrhagic leukoencephalitis: a critical entity for forensic pathologists to recognize. AB - Acute hemorrhagic leukoencephalopathy (AHLE) is a rare, acute disorder characterized by perivenular demyelination and diffuse hemorrhagic necrosis of the central nervous system. AHLE is thought to represent a hyperacute form of acute disseminated encephalomyelitis. AHLE is associated with a greater morbidity and mortality and, fortunately, is much less common than acute disseminated encephalomyelitis. Since most cases of AHLE result in patient demise, forensic pathologists should be cognizant of this entity and consider it in their differential diagnosis.Here we describe an interesting case of a previously healthy 11-year-old boy who initially complained of vague gastroenteritis-like symptoms while visiting a mountain lake. The boy's symptoms evolved to include severe headache and dizziness, necessitating a visit to a rural emergency department. He presented with focal neurologic findings, and head computed tomography (CT) scan confirmed thalamic edema. Cerebrospinal fluid analysis was suggestive of infectious etiology, and multiple empiric therapies were initiated. He was transferred to our institution, and his clinical status continued to worsen. Given the poor prognosis, the family requested withdrawal of supportive care. On day 14 of symptoms the boy succumbed to his illness. An autopsy was requested to further characterize the proximate cause of death.AHLE often presents with abrupt onset of fever, neck stiffness, seizure, and/or focal neurologic signs several days following a viral illness or vaccination. Thus, AHLE can clinically mimic a direct central nervous system infection or a toxic ingestion. AHLE has a very poor prognosis, with rapid deterioration and death usually occurring within days to one week after onset of symptoms. The cause for AHLE is unclear. An autoimmune pathophysiology is likely, with immune cross reactivity between myelin basic protein moieties and various infectious agent antigens. Treatment for AHLE is not well-established; some authors describe in recent literature that a combination of immunosuppressant medications and/or therapeutic plasma exchange may be of benefit in treating AHLE. PMID- 20010290 TI - Adolescent Russian roulette deaths. AB - Adolescence, between the ages of 10 and 19 years, is a unique period both physically and emotionally. During this time of life, individuals are known to experiment and engage in risky behavior, sometimes with unforeseen morbidity and mortality. We also see suicide emerge as a manner of death in this age group. The most common method is gunshot wound and sometimes in the form of Russian roulette. Few studies have looked at deaths by Russian roulette, the victims, and scenarios. In particular, no study examines the adolescent victim of Russian roulette. To better understand and classify this entity, adolescent Russian roulette autopsy cases over a 20-year period were examined looking at the victims, scenarios, autopsy findings, cause and manner of death, and the weapons. All victims were males, ages 13 to 19 years, with a Black-to-White ratio of 1:1. No victim had a previous psychiatric history. Toxicology was positive for alcohol and/or marijuana in 50% of the victims. Friends were present when the victim shot himself which occurred in the home the majority of the time. In all but 1 case, premeditation of the game was involved as the victim provided the weapon for the roulette. The cause of death was gunshot wound to the head (6 to the right side, 1 to the mouth, 1 to the forehead), and the manner of death was suicide in 6 cases and accident in 2 cases. A review of the literature discusses the adolescent victim, suicide, and Russian roulette. PMID- 20010291 TI - Danger assessment of a spring-gun device (muzzleloader) loaded only with black powder. AB - We report a case involving a spring-gun device (muzzleloader) loaded solely with gunpowder, installed next to shoes to prevent the neighbors' puppy from removing them. The booby trap was triggered by the 15-year-old dog-owners son when he tried to put the shoes out of the reach of the puppy. The boy suffered second degree superficial burns located mainly at the dorsal side of the right hand and fingers. To estimate the danger of the used weapon, several tests were undertaken on soap blocks from different distances and with different loads of black powder. The particle density per mm and the depth of black powder tattooing in the soap was compared with the boy's injuries, and found conclusive with the gun-owners statement regarding the loading of the weapon. Furthermore, our experiments indicated that the gunpowder load involved was not able to inflict permanent damage, not even to the eyes, at the here estimated firing distance. PMID- 20010292 TI - Postmortem imaging: MDCT features of postmortem change and decomposition. AB - Multidetector computed tomography (MDCT) has emerged as an effective imaging technique to augment forensic autopsy. Postmortem change and decomposition are always present at autopsy and on postmortem MDCT because they begin to occur immediately upon death. Consequently, postmortem change and decomposition on postmortem MDCT should be recognized and not mistaken for a pathologic process or injury. Livor mortis increases the attenuation of vasculature and dependent tissues on MDCT. It may also produce a hematocrit effect with fluid levels in the large caliber blood vessels and cardiac chambers from dependent layering erythrocytes. Rigor mortis and algor mortis have no specific MDCT features. In contrast, decomposition through autolysis, putrefaction, and insect and animal predation produce dramatic alterations in the appearance of the body on MDCT. Autolysis alters the attenuation of organs. The most dramatic autolytic changes on MDCT are seen in the brain where cerebral sulci and ventricles are effaced and gray-white matter differentiation is lost almost immediately after death. Putrefaction produces a pattern of gas that begins with intravascular gas and proceeds to gaseous distension of all anatomic spaces, organs, and soft tissues. Knowledge of the spectrum of postmortem change and decomposition is an important component of postmortem MDCT interpretation. PMID- 20010293 TI - Methanol intoxication: pathological changes of central nervous system (17 cases). AB - The nervous system has increased susceptibility for methanol intoxication. The aim of this study is to investigate various central nervous system lesions of methanol intoxication in 17 cases autopsied in the mortuary department of the Council of Forensic Medicine in Istanbul, Turkey. The reasons of methanol intoxication in the cases was likely the unwitting ingestion of methanol while drinking illegal alcohol. Survival times ranged from several hours to days. In 8 cases (47%), cerebral edema and in 9 cases (53%) at occipital, temporal and parietal cortex, basal ganglia and pons, petechial bleeding was observed. In addition to these findings, hemorrhagic necrosis were observed in thalamus, putamen, and globus pallidus in 5 cases (29.4%) and, in cerebral cortex in another 3 cases (17.6%). In 3 of the cases (17.6%) in which cerebral edema was found, herniation findings accompanied to the situation and in 2 cases (11.7%), pons bleeding was observed. Around the basal ganglia, in 2 of the cases with hemorrhagic necrosis, the situation ended with a ventricular compression. In 7 cases (41%), the associated findings of chronic ischemic changes in cortical neurons, lacunae formation, degeneration of granular cell layer of the cerebellum, and reactive gliosis were considered as the results of chronic alcoholism. PMID- 20010295 TI - Optimizing stem cell transplantation in myelodysplastic syndromes: unresolved questions. AB - PURPOSE OF REVIEW: Allogeneic hematopoietic stem cell transplantation (HCT) is the only curative therapy for myelodysplastic syndrome (MDS). Recent efforts to optimize the curative potential of transplant have focused on pretransplant therapy options, the use of predictive models to improve patient selection, and transplant modifications using reduced conditioning intensity. This review highlights strategies to optimize transplant for MDS and identifies unresolved questions. RECENT FINDINGS: Debate surrounding pretransplant therapy and HCT conditioning intensity for MDS continues. The current literature fails to identify a superior pretransplant treatment regimen; however, for treated patients achieving complete remission, the data suggest that myeloablative conditioning may not be required for successful transplant outcomes. Patient selection for transplant is also critical, and predictive tools (WHO classification-based prognostic scoring system and hematopoietic cell transplantation comorbidity index) have helped identify patients who may derive the most transplant benefit. SUMMARY: Prospective trials regarding optimal pretransplant therapy, utilization of patient selection tools, and conditioning intensity are warranted to improve transplant outcomes in MDS. Until those data are mature, current data suggest we initiate pretransplant therapy that minimizes toxicity and improves responses, try to optimize our patient selection using comorbidity (hematopoietic cell transplantation comorbidity index) and other predictive tools (WHO classification-based prognostic scoring system), and consider reduced intensity conditioning/nonmyeloablative conditioning in patients who have achieved a complete remission prior to transplant. PMID- 20010294 TI - Biology and clinical effects of natural killer cells in allogeneic transplantation. AB - PURPOSE OF REVIEW: Following allogeneic hematopoietic cell transplantation, donor derived natural killer (NK) cells target recipient hematopoietic cells, resulting in an antileukemia effect and a lower incidence of graft rejection. NK cells do not mediate and may diminish graft versus host disease. Here we review the determinants of NK cell alloreactivity and their implications for adoptive NK cell therapy. RECENT FINDINGS: NK cell alloreactivity has been defined by the absence of recipient MHC class I ligands for donor inhibitory killer immunoglobulin-like receptor (KIR) receptors, as predicted by a number of algorithms. Recently, the role of activating NK receptors and their cognate ligands has received more attention. The beneficial clinical effect of NK-cell alloreactivity has not been uniformly demonstrated, likely reflecting differences in conditioning regimens, graft components and posttransplant immune suppression. Investigations of NK cell phenotype and function after transplantation have helped demonstrate which NK cell subsets mediate the graft versus leukemia effect. These advances have proceeded in parallel with increasing facility in GMP grade bulk purification and administration of NK cell preparations. SUMMARY: NK cells are a heterogeneous population of lymphocytes with diverse patterns of target-cell recognition and effector function. Further clinical and functional correlations will help maximize their potential for clinical benefit. PMID- 20010296 TI - The role of exercise therapy in the management of juvenile idiopathic arthritis. AB - PURPOSE OF REVIEW: The current review summarizes the existing knowledge about exercise therapy in the management of juvenile idiopathic arthritis (JIA) along with activity level, functional abilities and exercise capacity of this population. RECENT FINDINGS: Current studies show that children with JIA are considerably less active than their peers. They have significantly impaired aerobic and anaerobic exercise capacity. The inactivity, decreased exercise capacity and disease course lead to deconditioning and disability. Adolescent girls with polyarticular rheumatoid factor-positive subtype appear to be most vulnerable to disability. Recent trials suggest that structured aerobic training or low-intensity programs do not exacerbate arthritis and can lead to improved physical fitness, quality of life and functional abilities in children and adolescents with JIA. SUMMARY: Inactivity in pediatric patients with JIA leads to deconditioning and disability and decreased bone mass, reduced quality of life and possibly increased mortality in adulthood. Although advances in pharmacology have improved the lives of children with JIA, management should also include a moderate, consistent exercise program or more active lifestyle. Physical activity may improve exercise capacity, decrease disability in adulthood, improve quality of life and, in some patients, decrease disease parameters. Further studies are needed to assess practicality of various programs and long-term effects of exercise in children and adolescents with JIA. PMID- 20010297 TI - Evidence supporting the use of physical modalities in the treatment of upper extremity musculoskeletal conditions. AB - PURPOSE OF REVIEW: To evaluate recent trials and reviews of physical modalities and conservative treatments for selected upper extremity musculoskeletal conditions for evidence supporting their use. RECENT FINDINGS: Recent evidence suggests that many localized tendinopathies are related more to degenerative than inflammatory processes. With this realization, there is increased emphasis on finding new modalities to treat tendinopathies and other localized musculoskeletal conditions that rely on other than anti-inflammatory mechanisms. Although there is good evidence to support the short-term benefits of corticosteroid injections, convincing evidence in support of other conservative treatments and modalities is generally lacking. Extracorpal shock wave therapy may have significant clinical benefit for calcific tendinitis; however, it requires intravenous sedation in most cases and does not appear to be effective in lateral epicondylitis. The most consistent positive treatment effects for rotator cuff tendinitis were achieved by ultrasound-guided subacromial corticosteroid injection as well as manual therapy in conjunction with therapeutic exercise. SUMMARY: Although there is evidence supporting the use of several different physical modalities and conservative treatments for upper extremity musculoskeletal conditions, there is a strong need for larger, higher quality randomized controlled trials. Although most studies are able to demonstrate short-term benefits, there is a lack of high-quality data demonstrating that these conservative treatments have long-term benefits, particularly, with regard to functional outcomes. PMID- 20010298 TI - Extrapyramidal signs predict the development of overt hepatic encephalopathy in patients with liver cirrhosis. AB - BACKGROUND/AIMS: The long-term evolution of cirrhotic patients with extrapyramidal signs has not yet been studied. We have investigated the influence of extrapyramidal signs on the prognosis, evolution, and quality of life of patients with liver cirrhosis. METHODS: Forty-six patients with cirrhosis were followed up and 18 of them were reevaluated, a mean of 45 months later. Cognitive impairment was measured with psychometric tests (Trail-Making Test part A, Grooved-Pegboard, Block-Design, Oral Symbol Digit and Stroop Test). Extrapyramidal signs were evaluated using the Unified Parkinson's Disease Rating Scale (UPDRS). Health-related quality of life was measured using the SF-36 scale and the Chronic Liver Disease Questionnaire. RESULTS: Eleven of the 46 patients who were followed up developed overt hepatic encephalopathy (HE) during the follow-up. The presence of extrapyramidal signs was the unique factor that predicted overt HE and patients with basal higher score in the part 3 of the UPDRS developed overt HE more frequently [hazard ratio=1.29; 95% confidence interval (1.04-1.60) P=0.023]. In the 18 reevaluated patients, there was an increase in the score of the UPDRS part 3 after follow-up. There was a worsening in the score values of the block design and the Number Connection Test. In health related quality of life scales, patients scored better in the area of mental health of the SF-36 scale. Patients with extrapyramidal signs persisted with worse scores in several items of the SF-36 scale and the Chronic Liver Disease Questionnaire. CONCLUSION: The presence of extrapyramidal signs in patients with liver cirrhosis predicts the development of overt HE. These signs increased along the follow-up, and remain a bad influence on quality of life. PMID- 20010299 TI - Amyloid deposition and influx transporter expression at the blood-brain barrier increase in normal aging. AB - Aging is the most important single risk factor for developing Alzheimer disease. We measured amyloid-beta peptide (Abeta) levels in rat cerebral cortex and hippocampus during normal aging of Brown-Norway/Fischer rats. Amyloid-beta accumulation was associated with expression of the Abeta influx transporter, the receptor for advanced glycation end-products (RAGEs) at the blood-brain barrier. Rats at selected ages from 3 to 36 months were analyzed by 1) immunohistochemistry for amyloid deposition and quantitative microvessel surface area RAGE expression, 2) ELISA for cortical Abeta40 and Abeta42 concentrations, and 3) Western blotting of microvessel proteins for RAGE expression. Immunohistochemistry showed increasing accumulation of brain Abeta with aging. By ELISA analysis, both Abeta40 and Abeta42 concentrations in cortical homogenates rose sharply from 9 to 12 months. The Abeta42 continued to rise up to age 30 months, whereas Abeta40 stabilized after 12 months. The expression of RAGE initially decreased between 3 and 12 months but then increased between 12 and 34 months by immunohistochemistry. On immunoblotting, RAGE decreased up to 9 months and then progressively increased up to 36 months. These data indicate an association between amyloid and microvessel RAGE during aging. An increase in capillary RAGE expression seems to play a role in the later Abeta accumulation but not in the initial increase. PMID- 20010300 TI - Development and characterization of a novel human in vitro blood-nerve barrier model using primary endoneurial endothelial cells. AB - There are phenotypic and functional differences between vascular endothelium from different tissues and between microvascular and macrovascular endothelial cells (ECs) from the same tissue. Relatively little is known about the human blood nerve barrier (BNB). We report the development of an in vitro BNB model using primary human endoneurial ECs freshly isolated and purified from decedent sciatic nerves via endoneurial stripping, connective tissue enzymatic digestion, and density centrifugation. Primary human endoneurial ECs are spindle shaped and contact inhibited. They rapidly differentiate to form capillary-like networks and microvessels, bind Ulex Europaeus Agglutinin 1 lectin, express von Willebrand factor, and endocytose acetylated low-density lipoprotein. They also express specific transport and cellular adhesion molecules and tight junction proteins, consistent with cells that form a highly restrictive endothelial barrier similar to the blood-brain barrier. When cultured on collagen-coated transwell inserts, the primary human endoneurial ECs develop an in vitro BNB with high transendothelial electrical resistances (160 Omega x cm(2); maximal 12 days after seeding) and low solute permeability coefficient to fluoresceinated high molecular weight (70 kDa) dextran (2.75 x 10(-3) cm/minute). This in vitro BNB model retains essential known or expected characteristics of the human BNB and has many potential applications for studies of solute, macromolecule, microbial, virus, and leukocyte interactions with this highly specialized endothelial barrier. PMID- 20010301 TI - Effects of antemortem and postmortem variables on human brain mRNA quality: a BrainNet Europe study. AB - Well-characterized and preserved human brain tissue that is prepared and stored in brain banks is an essential resource for research in neurological diseases. This study examined the quality of human brain postmortem tissue from multiple laboratories within the BrainNet Europe brain bank network to identify all possible confounding variables and determine how they may affect RNA quality. Antemortem and postmortem information was retrospectively collected for a large cohort of samples. Total RNA was isolated from anatomically defined brain regions using a standardized procedure; RNA quality was assessed using an Agilent 2100 Bioanalyzer. No significant difference in RNA quality was observed in 6 different brain regions. RNA quality deteriorated with increasing numbers of antemortem events such as hospitalization, coma, respiratory illness, and the use of artificial ventilation; accumulation of such events was associated with elevated hypoxia-inducible factor 1 alpha mRNA expression. Brain pH was found to be a good indicator of RNA quality. There was no correlation of postmortem delay with cerebrospinal fluid pH or RNA quality overall, but some individual RNAs decreased in quality with antemortem events and with postmortem delay. RNA quality did not affect total RNA yield. Determining the factors that are best predictors of RNA quality can help brain banks with selection criteria for storing high-quality brain tissue for research. PMID- 20010302 TI - Differential expression of CCN1/CYR61, CCN3/NOV, CCN4/WISP1, and CCN5/WISP2 in neurofibromatosis type 1 tumorigenesis. AB - The hallmark of neurofibromatosis type 1 is the development of dermal and plexiform neurofibromas. Neurofibromatosis type 1 patients with plexiform neurofibromas are at risk of developing malignant peripheral nerve sheath tumors. We applied a 22,000-oligonucleotide microarray transcriptomic approach to a series of plexiform neurofibromas in comparison with dermal neurofibromas, and results were confirmed with real-time quantitative reverse transcription polymerase chain reaction. Thirteen genes were upregulated and 10 were downregulated in plexiform neurofibromas. The upregulated genes mainly encode molecules involved in cell adhesion, extracellular matrix, fibrogenesis, and angiogenesis. Several CCN gene family members were dysregulated in neurofibromatosis type 1 tumorigenesis; the angiogenic gene CCN1/CYR61 was specifically upregulated in the plexiform neurofibromas; CCN4/WISP1 was upregulated, and CCN3/NOV and CCN5/WISP2 were downregulated in paired comparisons of plexiform neurofibroma and malignant peripheral nerve sheath tumor from the same patients. CCN1 and CCN3 proteins were detected by immunohistochemistry in neurofibromatosis type 1-associated tumors. Upregulation of S100A8, S100A9, and CD36 was also observed and suggests a role of this pathway in inflammation associated genesis of plexiform neurofibromas. In summary, a limited number of pathways are potentially involved in plexiform neurofibroma development. Some of the genes identified, particularly CCN1, might be useful diagnostic or prognostic markers or form the basis for novel therapeutic strategies. PMID- 20010304 TI - The effect of prolonged fixation time on immunohistochemical staining of common neurodegenerative disease markers. AB - The goals of this study were to determine the effects of prolonged fixation time and to optimize antigen retrieval (AR) methods on immunohistochemical (IHC) staining of common neurodegenerative disease markers. A panel of commercial antibodies (Abs) to amyloid-beta, ubiquitin, p62/sequestosome, tau, and alpha synuclein was applied to a 2-mm tissue microarray using several AR methods. The IHC outcomes were assessed in sections that included 2 types of specimens taken from 20 postmortem brains: short-term fixation of up to 70 days before paraffin embedment and long-term fixation of up to 14 years in formalin. Good amyloid-beta IHC staining was obtained with all amyloid-beta Abs applied when a formic acid AR method was used, even after 14 years of fixation. Ubiquitin immunoreactivity was also optimally labeled with this method. The p62/sequestosome IHC outcome was optimal for tissue fixed up to 10 years, but only when the p62-lck-ligand-Ab with heat AR method was used. All hyperphosphorylated tau Abs tested worked with fixation up to 10 years, in particular with the heat AR method, whereas Abs against tau isoforms RD3 and RD4 were applicable only when the fixation time was 6 months or shorter. alpha-Synuclein-immunoreactive structures were visualized up to 14 years but only by the use of Syn42-Ab after formic acid AR or after a combination of heat and formic acid methods. PMID- 20010303 TI - Clinical, neuropathologic, and biochemical profile of the amyloid precursor protein I716F mutation. AB - We report the clinical, pathologic, and biochemical characteristics of the recently described amyloid precursor protein (APP) I716F mutation. We present the clinical findings of individuals carrying the APP I716F mutation and the neuropathologic examination of the proband. The mutation was found in a patient with Alzheimer disease with onset at the age of 31 years and death at age 36 years and who had a positive family history of early-onset Alzheimer disease. Neuropathologic examination showed abundant diffuse amyloid plaques mainly composed of amyloid-beta42 and widespread neurofibrillary pathology. Lewy bodies were found in the amygdala. Chinese hamster ovary cells transfected with this mutation showed a marked increase in the amyloid-beta42/40 ratio and APP C terminal fragments and a decrease in APP intracellular domain production, suggesting reduced APP proteolysis by gamma-secretase. Taken together, these findings indicate that the APP I716F mutation is associated with the youngest age of onset for this locus and strengthen the inverse association between amyloid beta42/40 ratio and age of onset. The mutation leads to a protein that is poorly processed by gamma-secretase. This loss of function may be an additional mechanism by which some mutations around the gamma-secretase cleavage site lead to familial Alzheimer disease. PMID- 20010305 TI - Schwann cells producing matrix metalloproteinases under Mycobacterium leprae stimulation may play a role in the outcome of leprous neuropathy. AB - Matrix metalloproteinases (MMPs) mediate demyelination and breakdown of the blood nerve barrier in peripheral neuropathies. Matrix metalloproteinases and tissue inhibitor of metalloproteinase 1 gene expression and secretion were studied in cells of the human Schwann cell line ST88-14 stimulated with Mycobacterium leprae and tumor necrosis factor (TNF) and in nerve biopsies from patients with neural leprosy (n = 21) and nonleprous controls (n = 3). Mycobacterium leprae and TNF induced upregulation of MMP-2 and MMP-9 and increased secretion of these enzymes in cultured ST88-14 cells. The effects of TNF and M. leprae were synergistic, and anti-TNF antibody blockage partially inhibited this synergistic effect. Nerves with inflammatory infiltrates and fibrosis displayed higher TNF, MMP-2, and MMP-9 mRNA than controls. Leprous nerve biopsies with no inflammatory alterations also exhibited higher MMP-2 and MMP-9; tissue inhibitor of metalloproteinase 1 was significantly higher in biopsies with fibrosis and inflammation. Immunohistochemical double labeling of the nerves demonstrated that the MMPs were mainly expressed by macrophages and Schwann cells. The biopsies with endoneurial inflammatory infiltrates and epithelioid granulomas had the highest levels of MMP 2 and MMP-9 mRNA detected. Together, these results suggest that M. leprae and TNF may directly induce Schwann cells to upregulate and secrete MMPs regardless of the extent of inflammation in leprous neuropathy. PMID- 20010306 TI - TP53 mutation analysis of malignant peripheral nerve sheath tumors. AB - Mutations in TP53 underlie the development of malignant peripheral nerve sheath tumors (MPNSTs) in animal models, but there is controversy regarding the extent of TP53 mutations in human MPNSTs. We assessed the TP53 mutation frequency in 145 consecutive cases from our department over 36 years; 88 cases were histologically confirmed as MPNSTs, and corresponding clinical data were reviewed. Mutation analysis of TP53 Exons 4 to 9 on DNA from formalin-fixed, paraffin-embedded specimens was performed by bidirectional DNA sequencing. Tumors were localized in the extremities (n = 34), trunk (n = 34), or head and neck (n = 20). A minority of patients (n = 26, 30%) had neurofibromatosis type 1 (NF1); in these patients, the diagnosis of MPNST was made at younger ages (33 [SD, 3.6] years vs 49 [SD, 2.9] years in NF1 vs non-NF1; p = 0.003). High p53 protein expression was detected in 18 (21%) of 86 cases by immunohistochemistry. TP53 mutations were detected in 17 (24%) of 72 evaluable tumors, of which 36% were from NF1 patients. TP53 mutation and strong p53 immunostaining were positively correlated (p = 0.002); high proliferation indices correlated with cellular epithelioid and storiform growth patterns. These results indicate that TP53 mutations are relatively rare in human MPNST and that they are not positively correlated with the presence of NF1. PMID- 20010307 TI - Evolution of inflammation and white matter injury in a model of transient focal ischemia. AB - After an ischemic stroke, there is a prolonged inflammatory response and secondary phase of injury that is more amenable to treatment than acute neurotoxicity. Surprisingly, little is known about temporal and spatial relationships between inflammation and white matter injury. Here, we quantified development of white matter damage, inflammation, and a glial limitans at 1, 3, and 7 days after transient ischemia in the rat striatum using immunohistochemistry. Quantitative analysis showed that decreased staining for myelin basic protein and increased staining for damaged myelin basic protein began in the core, coincided with neutrophil infiltration, and progressed outward over time. Axon damage (i.e. accumulation of amyloid precursor protein) began at the edge of the lesion, coinciding with substantial microglia/macrophage activation, and progressed into the core. During the 7 days, activated microglia/macrophages dramatically increased only in the core and edge of the lesion. Detailed spatial analyses revealed that activated microglia/macrophages that surrounded undamaged axon bundles did not express ED1, a marker of phagocytic cells, whereas those inside damaged bundles expressed ED1. These results imply different contributions of neutrophils and microglia/macrophages to white matter injury after ischemic stroke. The distinct localizations of activated microglia/macrophages imply complex signals that regulate their migration toward and infiltration of damaged white matter. PMID- 20010308 TI - Predictable healing of femoral neck fractures treated with intraoperative compression and length-stable implants. AB - OBJECTIVES: A healed, yet shortened, femoral neck has historically been deemed a success in fracture treatment. This, however, comes at the price of diminished physical function and quality of life. We analyzed the outcomes of our treatment algorithm, which attempts to minimize postoperative shortening of femoral neck fractures and determined which preoperative factors were associated with femoral neck shortening and failure of surgical fixation. LEVEL OF EVIDENCE: This is level IV retrospective study. MATERIALS: Fifty-four patients underwent open reduction and internal fixation for acute femoral neck fracture with nonsliding constructs. The collapse of the femoral neck in the horizontal (X), vertical (Y), and along the resultant along the (Z) vector (X+Y=Z) was measured on anteroposterior radiographs corrected for leg rotation. The migration of the superior-most screw tip in all axes was measured. Age, gender, Garden grade, and Pauwel's angle were analyzed for their association with shortening or failure of surgical fixation. RESULTS: The average age of the patients was 78.1 years. There were 23 Garden I, 2 Garden II, 14 Garden III, and 15 Garden IV fractures. Fifty one (94%) healed successfully without complications. The minimum follow-up was 9 months (average, 17.6 months; range, 9-30 months). Surgical fixation failed in two patients, and one patient developed avascular necrosis. The average displacement of the femoral head and the screw tip was 1.23, 0.86, 1.98 mm and 0.7 mm, 0.9 mm, and 1.7 mm in the X, Y, and Z (resultant) vectors, respectively. DISCUSSION: With careful consideration to reduction, we fixed femoral neck fractures with nonsliding constructs, resulting in a high union rate with very minimal shortening of the femoral neck. PMID- 20010309 TI - Insomnia burden: definition and link to the menopausal transition. PMID- 20010310 TI - Neuroserpin is expressed in early stage of neurogenesis in adult rat hippocampus. AB - In the adult rat hippocampal formation, neurogenesis occurs in the dentate gyrus subgranular zone (SGZ). We used laser capture microdissection and an antidepressant-related genes microarray to analyze gene expression profiles of cells from the SGZ and from the outermost granule cell layer. Of the differentially expressed genes in the SGZ, we focused on neuroserpin, which is highly expressed in the adult rat SGZ. Neuroserpin immunoreactivity was present in cells positive for NeuN (postmitotic cell marker) and Tuj1 (immature neuron marker) but not in cells positive for calbindin (mature neuron marker). Although neuroserpin is expressed during late stage of neurogenesis in development, our results suggest that neuroserpin may play some roles in early stage of neurogenesis in adult rat hippocampus. PMID- 20010311 TI - Multidrug-resistant Enterococcus faecium meningitis in a toddler: characterization of the organism and successful treatment with intraventricular daptomycin and intravenous tigecycline. AB - A case of enterococcal meningitis in a toddler is presented. The organism was highly resistant to all drugs previously used for pediatric Gram-positive meningitis. She was successfully treated with intraventricular and intravenous daptomycin and intravenous tigecycline. The organism was characterized as a member of CC17, a notorious emerging nosocomial clone of Enterococcus faecium. PMID- 20010312 TI - Immunogenicity of a reduced schedule of pneumococcal conjugate vaccine in healthy infants and correlates of protection for serotype 6B in the United Kingdom. AB - BACKGROUND: Pneumococcal conjugate vaccine (PCV) was introduced in the United Kingdom immunization schedule in September 2006. This study was conducted to establish the immunogenicity of licensed PCV (Prevenar) at a reduced, 2 priming dose schedule (2+1) and to evaluate functional responses in the context of vaccine effectiveness. METHODS: Infants were randomized to receive PCV at 2 and 3 months or 2 and 4 months of age. Boosters were administered at the same time as Haemophilus influenzae type B/meningococcal C conjugate and Measles, Mumps and Rubella or with Measles, Mumps and Rubella alone (www.ClinicalTrials.gov NCT00197808). RESULTS: PCV at 2/3 months of age was poorly immunogenic and recruitment to this arm was terminated. PCV at 2/4 months of age resulted in lower than expected responses to serotypes 6B and 23F. Functional analysis of serotype 6B by OPA revealed that an enzyme-linked immunosorbent assay cutoff of 0.2 microg/mL was a better predictor of OPA positivity than a cut off of 0.35 microg/mL. PCV booster responses were excellent and no interference from concomitant vaccines was noted. CONCLUSIONS: An interval of at least 8 weeks is required when starting PCV vaccination at 2 months of age although not all serotypes are equally immunogenic. Correlates of protection derived from enzyme linked immunosorbent assay values may not be equally appropriate for all serotypes as illustrated by results for 6B in this study. PMID- 20010313 TI - Ventilation and oxygen: dose-related effects of oxygen on ventilation-induced lung injury. AB - Preterm infants are at high risk of developing ventilator-induced lung injury. We have used an animal model of in utero ventilation (IUV) to investigate the separate effects of ventilation and acute oxygen exposure on the very immature lung. Fetal sheep were ventilated in utero at 110 d gestation for 6 h with 100, 21, or 0% (100% nitrogen) oxygen (n = 5 each) and survived in utero, without further ventilation, until tissue collection at 118 d. Nonventilated 110 d and 118 d fetuses were used as controls. All IUV exposed fetuses had reduced secondary septal crest densities and increased elastin staining irrespective of the inspired oxygen concentration. IUV with 100% and 21% oxygen, but not 100% nitrogen, increased lung tissue volumes and myofibroblast differentiation and apoptosis within the distal lung parenchyma in a dose-dependent manner. This study shows that IUV without oxygen can reduce alveolarization, whereas ventilation with oxygen (6 h), even at levels found in air (21%), increases distal lung tissue volumes, elastin deposition, myofibroblast differentiation, and apoptosis. PMID- 20010314 TI - Resuscitation of hypoxic newborn piglets with supplementary oxygen induces dose dependent increase in matrix metalloproteinase-activity and down-regulates vital genes. AB - The optimal oxygen concentration for newborn resuscitation is still discussed. Oxygen administration during reoxygenation may induce short- and long-term pathologic changes via oxidative stress and has been associated to later childhood cancer. The aim was to study changes in oxidative stress-associated markers in liver and lung tissue of newborn pigs after acute hypoxia followed by reoxygenation for 30 min with 21, 40, or 100% oxygen compared with room air or to ventilation with 100% oxygen without preceding hypoxia. Nine hours after resuscitation, we found a dose-dependent increase in the matrix metalloproteinase gelatinase activity in liver tissue related to percentage oxygen supply by resuscitation (100% versus 21%; p = 0.002) pointing at more extensive tissue damage. Receiving 100% oxygen for 30 min without preceding hypoxia decreased the expression of VEGFR2 and TGFBR3 mRNA in liver tissue, but not in lung tissue. MMP , VEGF-, and TGFbeta-superfamily are vital for the development, growth, and functional integrity of most tissues and our data rise concern about both short- and long-term consequences of even a brief hyperoxic exposure. PMID- 20010315 TI - Association of the orphan nuclear receptor NR4A1 with tardive dyskinesia. AB - Recent evidence has identified the NR4A1 (NUR77, NGFI-B) gene as a strong candidate for involvement in tardive dyskinesia (TD). We have investigated the association of six single nucleotide polymorphisms within the NR4A family of genes with TD in a sample of 171 patients with schizophrenia of Caucasian descent. The NR4A1 single nucleotide polymorphism (SNP) marker rs2603751 showed a nominal association with the risk of TD, as well as with the extent of TD based on the Abnormal Involuntary Movements Scale (AIMS) scores. The haplotype generated by the markers rs2603751 and rs2701124 also showed association with TD and, after adjustment for multiple testing, both the NR4A1 marker rs2603751 and the haplotype continued to show a trend toward association with TD. Although the results of this study are limited by a small sample size, it presents important pilot data and warrants further investigation of the involvement of NR4A1 variants in TD. PMID- 20010316 TI - Polymorphic CA repeat in IGF-I gene: lack of association with schizophrenia. PMID- 20010317 TI - Maternal inheritance in recurrent early-onset depression. AB - Major depressive disorder (MDD) is believed to have a genetic factor in its pathogenesis. On the basis of studies in MDD showing brain energy depletion and maternal inheritance in some families, we hypothesize that some of the genetic factor is likely maternally inherited on the mitochondrial DNA (mtDNA). Six hundred and seventy-two pedigrees from the Genetics of Recurrent Early-Onset Depression project were analyzed for matrilineal/nonmatrilineal pairs. Pairs were constructed to control for sex, age and autosomal gene contribution (e.g. maternal vs. paternal aunts). Individuals with and without any mood disorder were tallied and compared across five different pairs. Matrilineal relatives (with the same mtDNA sequence as the proband) were significantly more likely to suffer from a mood disorder than were nonmatrilineal relatives (with another mtDNA sequence; odds ratio 2.0, 95% confidence interval: 1.5-2.6, P = 3 x 10(-6)). Our data show a modest maternal bias in the susceptibility towards the development of depression, suggesting that predisposing genetic factors likely reside on the mtDNA. Thus, our data strengthen the hypothesis that energy metabolism may be involved in the pathogenesis of depression. PMID- 20010318 TI - Meta-analysis of the association between the monoamine oxidase-A gene and mood disorders. AB - OBJECTIVE: To evaluate the controversial, putative associations between the three common polymorphisms [promoter variable number tandem repeat (uVNTR), T941G, (CA) repeat] of monoamine oxidase A (MAOA) and mood disorders (major depressive or bipolar disorders, BPD) by systematically meta-analyzing published case-control association studies. METHODS: We queried PubMed using the keywords 'MAOA', 'association' and 'depression' or 'bipolar'. Nine studies on uVNTR, seven studies on T941G, and eight studies on CA met the inclusion criteria. The meta-analysis was performed by sex and ethnicity. MAIN RESULTS: Our meta-analysis showed a significant association between uVNTR and MDD for the Asian group [odds ratio (OR) = 1.23 (1.02-1.47), P=0.03] and male Asian group [OR = 1.47 (1.06-2.05), P=0.02]. For the CA polymorphism, we found a significant association with BPD in the Caucasian group [OR = 1.28 (1.01-1.62), P=0.04] and female Caucasian group [OR = 1.36 (1.031-1.81), P=0.03]. For the CA polymorphism, we identified significant associations with BPD in all Caucasians for the overall alleles and for the specific alleles in a6 [OR = 1.35 (1.11-1.64), P=0.002] and in female Caucasians for the overall alleles and for the specific alleles in a2 [OR = 0.65 (0.48-0.90), P=0.009], a5 [OR = 1.44 (1.04-1.99), P=0.03], and a6 [OR = 1.41(1.12 1.78), P=0.004]. CONCLUSION: Our meta-analysis suggests a significant association of the MAOA gene with major depressive disorder and BPD within specific groups, indicating that these three polymorphisms of the MAOA gene may be associated with mood disorders by sex and ethnicity. Moreover, our systematic meta-analysis has revealed that although MAOA may be a common candidate gene for mood disorders, different polymorphisms and alleles appear to play different roles in major depressive disorder and BPD. PMID- 20010320 TI - Intravitreal bevacizumab for neovascular age-related macular degeneration with or without prior treatment with photodynamic therapy: one-year results. AB - PURPOSE: The purpose of this study was to elucidate the effect of prior photodynamic therapy (PDT) on the efficacy of intravitreal bevacizumab for the treatment of neovascular age-related macular degeneration. METHODS: One hundred and nine eyes of 102 patients with neovascular age-related macular degeneration were evaluated-80 eyes without prior treatment (group 1) and 29 with prior PDT (group 2). Best-corrected visual acuity, ocular coherence tomography, and funduscopy were assessed monthly. Results were evaluated at 1, 3, 6, and 12 months. RESULTS: One hundred and one eyes completed a 12-month evaluation. At 12 months, best-corrected visual acuity increased 5.6 letters with treatment (P = 0.001): +5.7 letters in group 1 and +5.4 in group 2 (P = 0.92). Overall, visual acuity improved > or =15 letters in 22.5% of eyes: 24.0% of naive eyes versus 18.5% with prior PDT (P = 0.56). Best-corrected visual acuity loss > or =15 letters occurred in 6 eyes, 5 with naive lesions. An overall reduction in ocular coherence tomography central retinal thickness was observed at all time points. Mean number of injections per eye per year was 5.6, 6.13 in group 1 versus 4.22 in group 2 (P = 0.01). Two retinal pigment epithelial tears, one subretinal macular hemorrhage, and two strokes occurred in naive lesions. CONCLUSION: The authors showed similar efficacy for intravitreal bevacizumab independently of prior PDT treatment. Eyes with prior PDT needed a statistically significantly lower number of injections to control their lesions. PMID- 20010321 TI - Retinal pigment epithelial changes in chronic Vogt-Koyanagi-Harada disease: fundus autofluorescence and spectral domain-optical coherence tomography findings. AB - PURPOSE: The purpose of this study was to determine whether fundus autofluorescence (FAF) and spectral domain-optical coherence tomography (SD-OCT) imaging allow better assessment of retinal pigment epithelium and the outer retina in subjects with chronic Vogt-Koyanagi-Harada disease compared with examination and angiography alone. METHODS: A cross-sectional analysis of a series of seven consecutive patients with chronic Vogt-Koyanagi-Harada disease undergoing FAF and SD-OCT was conducted. Chronic disease was defined as duration of intraocular inflammation >3 months. Color fundus photographs were correlated to FAF and SD-OCT images. The images were later correlated to fluorescein angiography and indocyanine green angiography. RESULTS: All patients had sunset glow fundus, which resulted in no apparent corresponding abnormality on FAF or SD OCT. Lesions with decreased autofluorescence signal were observed in 11 eyes (85%), being associated with loss of the retinal pigment epithelium and involvement of the outer retina on SD-OCT. In 5 eyes (38%), some of these lesions were very subtle on clinical examination but easily detected by FAF. Lesions with increased autofluorescence signal were seen in 8 eyes (61.5%), showing variable involvement of the outer retina on SD-OCT and corresponding clinically to areas of retinal pigment epithelium proliferation and cystoid macular edema. CONCLUSION: Combined use of FAF and SD-OCT imaging allowed noninvasive delineation of retinal pigment epithelium/outer retina changes in patients with chronic Vogt-Koyanagi-Harada disease, which were consistent with previous histopathologic reports. Some of these changes were not apparent on clinical examination. PMID- 20010322 TI - Intravitreal bevacizumab injection for central serous chorioretinopathy. AB - PURPOSE: The purpose of this study was to evaluate the effectiveness of intravitreal injection of bevacizumab for treatment of central serous chorioretinopathy. METHODS: In this retrospective case series, six patients (six eyes) with central serous chorioretinopathy were treated with an intravitreal injection of bevacizumab. The outcome measures included visual acuity with Early Treatment Diabetic Retinopathy Study letters, central macular thickness measurement with optical coherence tomography, changes in fluorescein angiography, and indocyanine green angiography. RESULTS: The mean age of the patients was 42.3 years and the mean follow-up period was 9.0 months (range, 5-12 months). Mean visual acuity +/- standard deviation increased from 40.8 +/- 8.3 Early Treatment Diabetic Retinopathy Study letters at baseline to 49.0 +/- 5.0 Early Treatment Diabetic Retinopathy Study letters at 1 month (P = 0.046) and to 53.3 +/- 5.2 Early Treatment Diabetic Retinopathy Study letters at 3 months (P = 0.028). Mean central macular thickness +/- standard deviation decreased from 331.5 +/- 93.4 microm to 164 +/- 34 microm at 3 months (P = 0.043). Leakage on fluorescein angiography and hyperpermeability on indocyanine green angiography decreased in conjunction with improvement in central macular thickness observed by optical coherence tomography. CONCLUSION: Intravitreal bevacizumab injections resulted in improved visual acuity and anatomical results for central serous chorioretinopathy. PMID- 20010323 TI - Association of positive dual-modality positron emission tomography/computed tomography imaging of primary choroidal melanoma with chromosome 3 loss and tumor size. AB - PURPOSE: To evaluate the positive dual-modality positron emission tomography/computed tomography (PET/CT) of choroidal melanoma with chromosome 3 loss and tumor size. METHODS: Thirty-seven consecutive patients with choroidal melanoma with known chromosome 3 status who underwent whole-body PET/CT imaging were retrospectively reviewed. Cytology and chromosome 3 loss were identified by fine-needle aspiration biopsy. Fluorescent in situ hybridization and whole genome microarray by single-nucleotide polymorphism were used to evaluate the chromosome 3 status. Metabolic activity of primary choroidal melanoma by PET/CT imaging was evaluated. RESULTS: Thirteen of 37 (35%) primary choroidal melanomas had loss of chromosome 3; 7 of the 13 (54%) melanomas were positive for metabolic activity identified by PET/CT imaging. All 24 of 37 melanomas without chromosome 3 loss were inactive for metabolic activity. There was a statistically significant association between positive metabolic activity and chromosome 3 loss (P = 0.00017 Fisher exact test); positive PET/CT imaging was 54% sensitive and 100% specific for loss of chromosome 3. Seven of 37 (19%) choroidal melanomas with positive metabolic activity by PET/CT were statistically significantly larger in size than the 30 metabolically inactive melanomas (P < 0.001, Kruskal-Wallis test). CONCLUSION: Positive metabolic activity of choroidal melanoma identified by PET/CT imaging was statistically significantly associated with chromosome 3 loss and larger tumor size. PMID- 20010324 TI - Infliximab stability after reconstitution, dilution, and storage under refrigeration. AB - PURPOSE: The purpose of this study was to investigate the stability of reconstituted infliximab solutions and determine whether infliximab is suitable for compounding for potential intravitreal use. METHODS: Infliximab was reconstituted, and the solution was aliquoted and stored refrigerated. On each day of testing, an aliquot was serially diluted to concentrations ranging from 50,000 pg/mL to 69 pg/mL. Each dilution was assayed by microsphere immunoassay daily for 5 days and weekly for a total of 6 weeks. The outcome measure was median fluorescence intensity measured by dual laser flow analysis of fluorochrome-labeled secondary antibodies to infliximab bound to tumor necrosis factor-alpha-coated microspheres. RESULTS: There was an increasing median fluorescence intensity for increasing infliximab concentration in a sigmoidal dose-response curve with a variable slope that was equivalent for each time point. Each respective concentration of infliximab showed nearly equivalent median fluorescence intensity for every time point over the 6-week period. CONCLUSION: The authors found that the immunoreactivity of 2 different concentrations of infliximab stored at 4 degrees C over a 6-week period remained stable. Infliximab is suitable for compounding and could be a cost-effective intravitreal medication for use in clinical practice if further study supports its safety and efficacy. PMID- 20010325 TI - Minimal fluid-air exchange in combined 23-gauge sutureless vitrectomy, phacoemulsification, and intraocular lens implantation. AB - PURPOSE: The purpose of this study was to compare rates of postoperative hypotony and intraocular lens-related complications between minimal fluid-air exchange and partial fluid-air exchange in combined 23-gauge vitrectomy and cataract surgery. METHODS: A prospective, consecutive, interventional case series of 48 eyes that underwent 23-gauge vitrectomy with a minimal fluid-air exchange (minimal F-A group) was compared with a retrospective, consecutive case series of 38 eyes that underwent 23-gauge vitrectomy with a partial fluid-air exchange (partial F-A group). The main outcome measures were postoperative hypotony (<6 mmHg) and intraocular lens-related complications, such as posterior capsule opacification or pupillary capture. RESULTS: Two (5.3%) of 38 eyes in the partial F-A group had hypotony, and only 1 (2.1%) of 48 eyes in the minimal F-A group had hypotony (P > 0.05). Posterior capsule opacification was identified in 11 (28.9%) of 38 eyes in the partial F-A group but only in 4 (8.3%) of 48 eyes in the minimal F-A group (P = 0.013). Pupillary capture was observed in 3 (7.9%) of 38 eyes in the partial F A group, but it was absent in the minimal F-A group (P = 0.049). No retinal detachment or endophthalmitis was developed in both groups during follow-up. CONCLUSION: Minimal fluid-air exchange in combined 23-gauge sutureless vitrectomy and cataract surgery may reduce postoperative hypotony and intraocular lens related complications. PMID- 20010326 TI - Immune cell functional assay in monitoring of adult liver transplantation recipients with infection. AB - PURPOSE: To identify the levels of functional immunity measured by the ImmuKnow assay in Chinese liver transplantation recipients and its application in monitoring the risk of posttransplant infection. METHODS: Forty-five apparent healthy Chinese and 106 adult liver transplant (LT) recipients were under investigation. LTs were grouped in stable status or infection according to their clinical diagnosis. Whole blood samples were collected freshly and cultured within 6 hr, the CD4(+) T cells were selected, and their adenosine triphosphate (ATP) value was assayed the next day. Before stimulation, we also examined the percentage of T-helper (Th; CD3(+) CD4(+)) and T-suppress (Ts; CD3(+) CD8(+)) lymphocyte subpopulations and the ratio of Th/Ts. RESULTS: The average ImmuKnow assay in infectious LT recipients was 128 + or - 84 ng/mL, significantly lower (P<0.05) than that in stable LTs (305 + or - 149 ng/mL) or in normal adults (301+ or - 101 ng/mL). The ImmuKnow values in LTs had a good negative correlation to infection clinically (r = -0.6217, P<0.001). Infectious risk was high when the ImmuKnow value was less than 130 ng/mL (odds ratio=13, 95% confidence interval 6.0-29.4, P<0.01). The sensitivity of low ImmuKnow values in posttransplant infection was 85.2%, significantly higher than those of Th/Ts ratio and immunosuppressant trough levels (P<0.01); specificity was 76.3%, comparable with that of Th/Ts ratio (75.5%), but greatly higher than immunosuppressant trough levels (P<0.01). ImmuKnow ATP values had no correlation with Th/Ts ratio or immunosuppressant trough levels. CONCLUSION: ImmuKnow ATP levels are lower in LT recipients with infection, which provides a new tool in monitoring posttransplant infection, and an index of tailoring immunosuppression clinically. PMID- 20010327 TI - Steroid-free maintenance immunosuppression and ABO-incompatible transplantation. PMID- 20010328 TI - Late steroid withdrawal after ABO blood group-incompatible living donor kidney transplantation: high rate of mild cellular rejection. AB - BACKGROUND: Little is known about the safety of steroid withdrawal after ABO blood group-incompatible living donor kidney transplantation. METHODS: Between September 2005 and November 2007, a total of 15 patients were successfully transplanted over the blood group barrier in our transplantation centers. Similarly to transplant recipients with normal immunological risk, we aimed to taper and eventually stop oral prednisone because of the well-known negative impact of steroids on cardiovascular morbidity and mortality, which in turn is limiting graft survival. RESULTS: Up to now, patient and graft survival is 100% after a median follow-up of 839 days (range, 513-1281 days). On the basis of serial protocol biopsies, late steroid withdrawal could successfully be performed in only 5 of 11 patients. Nevertheless, the remaining 6 patients showed histologic signs of mild and subclinical acute rejection shortly after complete withdrawal or even during steroid tapering. CONCLUSIONS: With this elevated risk of at least subclinical acute rejection after late steroid withdrawal, we propose that steroid withdrawal in ABO blood group-incompatible kidney graft recipients should only be performed after a protocol biopsy showing normal tissue and together with a thorough clinical and in doubtful cases also histologic follow up. PMID- 20010329 TI - Outcome of renal allograft in patients with Henoch-Schonlein nephritis: single center experience and systematic review. AB - BACKGROUND: Henoch-Schonlein nephritis (HSN) is a rare condition resulting in end stage renal disease. Therefore, graft outcomes and recurrence rates after transplantation are not well studied. Also, the effect of donor type on graft outcome has not been evaluated thoroughly. METHODS: The graft outcome and recurrence rate in 20 kidney recipients with HSN were compared with age-, sex-, and donor source-matched controls (control A, primary immunoglobulin A nephropathy; control B, other causes; 40 recipients per group). To assess the effect of donor type, we pooled our data with two previous cohort studies where donor type had been described in detail. RESULTS: Overall graft survival rates were 87.7% at 10 years. The overall recurrence rate of HSN was 15.4% over 10 years. Graft survival and recurrence rates in the HSN group were similar to those of control A and control B. The pooled data showed a 29.4% incidence rate for recurrent HSN. Living related donor transplantation showed a trend of higher recurrence compared with recipients with nonrelated grafts, although it was marginally significant (P=0.059). However, the graft survival rate in related donor recipients was not inferior to that in the unrelated-donor recipients. CONCLUSIONS: Long-term graft survival and recurrence rates in kidney recipients with HSN were comparable to those of recipients with primary immunoglobulin A nephropathy. The type of donor did not significantly affect long-term graft survival. PMID- 20010330 TI - Angiotensin blockade is associated with early graft dysfunction after live donor renal transplantation. AB - BACKGROUND: Activation of the renin angiotensin system may contribute to the development of delayed graft function after renal transplantation. We, therefore, examined the impact of pretransplant treatment with blockers of the renin angiotensin system on early graft function after renal transplantation. METHOD: The case records of all patients who received a live donor transplant between January 2001 and August 2008 were reviewed. RESULTS: Serum creatinine measurements were recorded for the first nine posttransplant days, in 94 recipients of live donor kidneys, where there were neither surgical nor hemodynamic complications and no documented rejection or infection. Forty patients (43%) were treated with a blocker of the renin-angiotensin-aldosterone system before transplantation. Serum creatinine levels fell more slowly in these patients (P=0.02). Two patients required hemodialysis after transplantation; both were on an angiotensin-converting enzyme inhibitor/angiotensin II receptor blocker. CONCLUSION: Live donor renal transplant recipients who are taking a renin-angiotensin-aldosterone system blocker at the time of transplantation are more likely to have impaired graft function postoperatively in the absence of other explanations. This observation requires further exploration in live and deceased donor renal transplantation. PMID- 20010331 TI - Quality of life and symptoms of anxiety and depression of patients receiving cancer chemotherapy: longitudinal study. AB - The overall survival gain of cancer chemotherapy is estimated to be small, and some claim that it rarely improves quality of life (QOL). The purpose of this population-based study was to describe QOL and symptoms of anxiety and depression, over time, in a group of Icelandic cancer patients receiving chemotherapy. Quality of life was measured with the Icelandic version of Cancer Rehabilitation Evaluation System, Short Form (CARES-SF), and symptoms of anxiety and depression, with the Icelandic version of Hospital Anxiety and Depression Scale (HADS). Enrolled into the study were 144 patients, 90 women and 54 men; mean (SD) age was 55 (12.1) years. Although QOL was found relatively good during the period of chemotherapy, it was significantly worse after 3 and 6 months compared to baseline. Quality of life was found worst in the sexual and physical domains. At all time points, a significant association was found between symptoms of anxiety and depression and QOL with those reporting symptoms of either anxiety or depression experiencing worse QOL. The good QOL found during the period of chemotherapy is a positive finding. The results, however, call for an intense assessment, over time, of physical symptoms, anxiety, and depression, as well as sexuality issues. PMID- 20010332 TI - Helping another die well: is this a task of human development? PMID- 20010333 TI - Cardiac amyloidosis: approaches to diagnosis and management. AB - Amyloidosis is a clinical disorder caused by the extracellular deposition of misfolded, insoluble aggregated protein with a characteristic ss pleated sheet configuration that produces apple-green birefringence under polarized light when stained with Congo red dye. The spectrum of organ involvement can include the kidneys, heart, blood vessels, central and peripheral nervous systems, liver, intestines, lungs, eyes, skin, and bones. Cardiovascular amyloidosis can be primary, a part of systemic amyloidosis, or the result of chronic systemic disease elsewhere in the body. The most common presentations are congestive heart failure because of restrictive cardiomyopathy and conduction abnormalities. Recent developments in imaging techniques and extracardiac tissue sampling have minimized the need for invasive endomyocardial biopsy for amyloidosis. Cardiac amyloidosis management will vary depending on the subtype but consists of supportive treatment of cardiac related symptoms and reducing the amyloid fibrils formation attacking the underlying disease. Despite advances in treatment, the prognosis for patients with amyloidosis is still poor and depends on the underlying disease type. Early diagnosis of cardiac amyloidosis may improve outcomes but requires heightened suspicion and a systematic clinical approach to evaluation. Delays in diagnosis, uncertainties about the relative merits of available therapies, and difficulties in mounting large-scale clinical trials in rare disorders combine to keep cardiac amyloidosis a challenging problem. This review outlines current approaches to diagnosis, assessment of disease severity, and treatment of cardiac amyloidosis. PMID- 20010334 TI - Clinical applications of highly sensitive troponin assays. AB - Cardiac troponin is the biomarker of choice for the diagnosis of acute myocardial infarction. Recent consensus recommendations have adopted a concentration of troponin above the 99th percentile of a healthy population to diagnose myocardial infarction. Until recently, there was no assay capable of achieving recommended precision; however, with the development of "highly sensitive" troponin assays, it is now possible to accurately measure troponin concentrations at and below the current 99th percentile of a healthy population. These assays have enormous potential in not only identifying more patients with acute myocardial infarction, and providing superior risk prediction in those so afflicted, in addition highly sensitive troponins assays may be useful for long-term risk assessment of the patient with coronary disease. In this article, we will review the clinical applications, novel concepts, challenges, and limitations of using highly sensitive troponins assays. PMID- 20010335 TI - Assessment of vascular endothelial function with peripheral arterial tonometry: information at your fingertips? AB - Endothelial dysfunction is an important component in the pathogenesis of atherosclerosis. The ability to assess the endothelium in a meaningful manner has been the subject of intense investigation over decades. Since the function of endothelial cells is a gauge of vascular health, assessment of vascular function is emerging as a useful tool for predicting cardiovascular risk and as a surrogate outcome measure for cardiovascular reduction intervention studies. This review highlights techniques for assessing endothelial function, focusing on a novel method of determining peripheral vascular reactivity via arterial tonometry. PMID- 20010336 TI - Critical appraisal on the utility of echocardiography in the management of acute pulmonary embolism. AB - Echocardiography can be used for rapid and accurate risk stratification of patients with pulmonary embolism to appropriately direct the therapeutic strategies for those at high risk. Echocardiography is an ideal risk stratification tool in this regard because of its easy portability to the emergency room or to the bed side. It can be performed at a relatively low cost and at no risk to the patient. Furthermore, echocardiography allows repetitive noninvasive assessment of the cardiovascular and hemodynamic status of the patient and the response to the therapeutic interventions. Right ventricular hypokinesis, persistent pulmonary hypertension, a patent foramen ovale, and a free floating right heart thrombus are echocardiographic markers that identify patients at a higher risk for morbidity and mortality. Such patients warrant special consideration for thrombolysis or embolectomy. PMID- 20010337 TI - Atrial fibrillation in congestive heart failure. AB - Atrial fibrillation (AF) and heart failure (HF) are common and interrelated conditions, each promoting the other, and both associated with increased mortality. HF leads to structural and electrical atrial remodeling, thus creating the basis for the development and perpetuation of AF; and AF may lead to hemodynamic deterioration and the development of tachycardia-mediated cardiomyopathy. Stroke prevention by antithrombotic therapy is crucial in patients with AF and HF. Of the 2 principal therapeutic strategies to treat AF, rate control and rhythm control, neither has been shown to be superior to the other in terms of survival, despite better survival in patients with sinus rhythm compared with those in AF. Antiarrhythmic drug toxicity and poor efficacy are concerns. Catheter ablation of AF can establish sinus rhythm without the risks of antiarrhythmic drug therapy, but has important procedural risks, and data from randomized trials showing a survival benefit of this treatment strategy are still lacking. In intractable cases, ablation of the atrioventricular junction and placement of a permanent pacemaker is a treatment alternative; and biventricular pacing may prevent or reduce the negative consequences of chronic right ventricular pacing. PMID- 20010340 TI - Current Opinion in Anaesthesiology. Current world literature. PMID- 20010338 TI - Hydrochlorothiazide versus chlorthalidone in the management of hypertension. AB - Thiazide diuretics are a mainstay for the treatment of hypertension. Although there are several thiazide diuretics currently available, hydrochlorothiazide (HCTZ) continues to be the most popular thiazide used for treating high blood pressure. This is despite several clinical trials that have used and documented the benefits of chlorthalidone for hypertension management. In terms of blood pressure lowering, both HCTZ and chlorthalidone appear to be very effective. Head to-head studies have shown trends favoring chlorthalidone as a more effective blood pressure lowering agent compared with HCTZ, but statistical significance in this regard has not been consistently demonstrated. Also unclear is the relative benefits of these 2 drugs with regards to reducing clinical complications of hypertension, namely cardiovascular morbidity and mortality. Whereas there is more aggregate clinical trial data documenting the benefits of chlorthalidone compared with HCTZ in terms of hard clinical outcomes, these trials are not direct comparisons and can only be used to fuel the debate as to which of the 2 thiazides are better. From a safety perspective, hypokalemia is a risk with any of the thiazide diuretics and has been shown to be dose-related. However, at equipotent dosages, the incidence of hypokalemia between chlorthalidone and HCTZ appears comparable. The available evidence therefore supports both HCTZ and chlorthalidone as safe and effective drugs for treating hypertension. Although there are favorable trends both in terms of antihypertensive efficacy as well as clinical outcomes data with chlorthalidone compared with HCTZ, the results are not conclusive, and as such may not be enough to shift the treatment paradigm in favor of chlorthalidone, given the comfort level that most prescribers have with HCTZ. A head-to-head study looking at hard clinical outcomes, which may or may not ever be performed, may be the only way to resolve the ongoing debate as to which is the preferred thiazide for treating hypertension. PMID- 20010341 TI - We take our pulse. PMID- 20010342 TI - Prevention of metachronous colorectal cancer in patients with Lynch syndrome. PMID- 20010343 TI - Evaluating the age distribution of patients with colorectal cancer: are the United States Preventative Services Task Force guidelines for colorectal cancer screening appropriate? AB - PURPOSE: Evaluate the appropriateness of the 2008 United States Preventative Services Task Force screening recommendations for colorectal cancer. METHODS: Ages at diagnosis data were collected on patients with colorectal cancer from the William Beaumont Tumor Registry. The database identified 6,925 patients treated for colorectal cancer between January 1973 and December 2007. Patients were divided into 3 age groups at diagnosis categories (younger than 50 years old, those 50 to 75 years old, and those older than 75 years old) to evaluate whether there were changes in the age distribution, pathologic stage, or tumor location during the 35-year period. RESULTS: The percent of patients with colorectal cancer older than age 75 years increased from 29% to 40% (P < .0001). The combined percentage of patients younger than age 50 years and older than age 75 years has increased from 36% (1973-1978) to 49% (2003-2007). The combined percentages of stage III and IV disease in patients younger than 50 years and older than 75 years were 51% and 34%, respectively (P < .0001). Rectal or left sided lesions occurred in 68%, 64%, and 50% of patients younger than 50 years old, those 50 to 75 years old, and those older than 75 years old, respectively (P < .0001). Right-sided lesions occurred in 22%, 25%, and 37% of those younger than 50 years old, those 50 to 75 years old, and those older than 75 years old, respectively (P < .0001). CONCLUSIONS: There has been a significant increase in the percentage of colorectal cancer patients older than age 75 years. Failing to screen patients younger than 50 years and older than the age of 75 years would miss 49% of patients treated at our institution from 2003 to 2007. PMID- 20010344 TI - Effect of race on colon cancer treatment and outcomes in the Department of Defense healthcare system. AB - PURPOSE: The increase in mortality noted in African Americans with colon cancer is attributed to advanced stage at presentation and disparities in treatment received. The aim of this study was to assess the influence of race on the treatments and survival of colon cancer patients in an equal-access healthcare system. METHODS: This retrospective cohort study included African American and white patients with colon cancer treated at Department of Defense facilities. Disease stage, surgery performed, chemotherapy used, and overall survival were evaluated. RESULTS: Of the 6958 colon cancer patients identified, 1115 were African American. African Americans presented more frequently with stage IV disease, 23% vs 17% for whites (P < .001). There was no difference in surgical resection rates for African American or whites (85.8% vs 85.5%, respectively; chi2, P > .05). There was no difference in the use of systemic chemotherapy for stage III colon cancer (73.5% for African Americans vs 72.2% for whites; chi2, P > .05) or stage IV colon cancer (56.3% for African Americans vs 54.4% for whites; chi2, P > .05). The overall 5-year survival rate was similar for African American and white patients (56.1% vs 58.5%, respectively; log-rank, P > .05). After adjusting for gender, age, tumor grade, and stage, African American race was not a risk factor for survival in Cox proportional hazard analysis (hazard ratio, 0.981; 95% confidence interval, 0.888-1.084). CONCLUSIONS: In an equal-access healthcare system, African American race is not associated with an increase in mortality. African American patients undergo surgery and chemotherapy is administered at rates equal to whites for all stages of colon cancer. PMID- 20010345 TI - Transanal endoscopic microsurgery resection of rectal tumors: outcomes and recommendations. AB - PURPOSE: Transanal endoscopic microsurgery provides a minimally invasive alternative to radical surgery for excision of benign and malignant rectal tumors. The purpose of this study was to review our experience with transanal endoscopic microsurgery to clarify its role in the treatment of different types of rectal pathology. METHODS: A prospective database documented all patients undergoing transanal endoscopic microsurgery from October 1996 through June 2008. We analyzed patient and operative factors, complications, and tumor recurrence. For recurrence analysis, we excluded patients with fewer than 6 months of follow up, previous excisions, known metastases at initial presentation, and those who underwent immediate radical resection following transanal endoscopic microsurgery. RESULTS: Two hundred sixty-nine patients underwent transanal endoscopic microsurgery for benign (n = 158) and malignant (n = 111) tumors. Procedure-related complications (21%) included urinary retention (10.8%), fecal incontinence (4.1%), fever (3.8%), suture line dehiscence (1.5%), and bleeding (1.5%). Local recurrence rates for 121 benign and 83 malignant tumors were 5% for adenomas, 9.8% for T1 adenocarcinoma, 23.5% for T2 adenocarcinoma, 100% for T3 adenocarcinoma, and 0% for carcinoid tumors. All 6 (100%) recurrent adenomas were retreated with endoscopic techniques, and 8 of 17 (47%) recurrent adenocarcinomas underwent salvage procedures with curative intent. CONCLUSIONS: Transanal endoscopic microsurgery is a safe and effective method for excision of benign and malignant rectal tumors. Transanal endoscopic microsurgery can be offered for (1) curative resection of benign tumors, carcinoid tumors, and select T1 adenocarcinomas, (2) histopathologic staging in indeterminate cases, and (3) palliative resection in patients medically unfit or unwilling to undergo radical resection. PMID- 20010346 TI - An increase in compliance with the Surgical Care Improvement Project measures does not prevent surgical site infection in colorectal surgery. AB - PURPOSE: The primary goal of the Surgical Care Improvement Project is to improve quality of care by implementing evidence-based health care practices that prevent surgical complications. This study was designed to test the hypothesis that an increase in compliance with quality process measures decreases the rate of surgical site infections in patients undergoing colorectal surgeries. METHODS: A multidisciplinary task force implemented and monitored compliance with individual quality measures in patients undergoing elective colorectal resections at a tertiary institution. Individual compliance rates and infections were collected prospectively and reviewed monthly. For data analysis, patients were assigned to 2 consecutive 14-month periods: period A (April 1, 2006 to May 31, 2007) and period B (June 1, 2007 to July 31, 2008). Comparisons between periods were performed to determine the association of compliance with process measures and outcomes in infections. RESULTS: A total of 491 consecutive patients were treated during the study periods (period A: n = 238; period B: n = 253). There were no statistically significant differences in patient characteristics, diagnoses, or surgical procedures between periods. Compliance with all process measures significantly increased within periods except for perioperative glucose control. Global compliance (compliance with all measures per patient) significantly improved from period A to B (40%-68%, respectively; P < .001). In total, 99 patients (19%) developed surgical site infections (period A, 18.9%; period B, 19.4%). CONCLUSION: An increase in compliance with the Surgical Care Improvement Project aimed to prevent surgical site infections does not translate into a significant reduction of surgical site infections in patients undergoing colorectal resections. PMID- 20010347 TI - Cancer/testis antigens and clinical risk factors for liver metastasis of colorectal cancer: a predictive panel. AB - PURPOSE: Accumulating evidence suggests that cancer/testis antigens may serve as indicators of tumor malignant phenotype. The purpose of this study is to evaluate cancer/testis antigen genes in predicting metastasis of colorectal cancer to the liver. METHODS: The expression levels of 25 cancer/testis antigen genes were determined by reverse-transcription polymerase chain reaction in 288 colorectal cancer tissue samples from the primary tumor or liver metastasis. Pearson chi2 and multiple logistic regression analyses were performed to assess the association between risk factors and probability of liver metastasis of colorectal cancer. RESULTS: No significant difference was detected between the primary tumor and liver metastasis in expression pattern of cancer/testis antigen genes in colorectal cancer tissue samples. However, 3 cancer/testis antigen genes (PAGE4, SCP-1, and SPANX) and 3 clinicopathologic parameters (lymph node involvement, vessel cancer embolus, and tumor invasion depth) correlated significantly with liver metastasis of colorectal cancer (P < .05). A logistic regression model was constructed for prediction of liver metastasis based on a panel consisting of PAGE4, lymph node involvement, and presence or absence of vessel cancer embolus. The predicted risk of liver metastasis based on the panel was consistent with the actual risk observed. The probability of developing liver metastasis as estimated by the panel was 86.9% when all 3 factors were positive, representing an up to 20% improvement in the prediction level compared with the classic methods of lymph node involvement and vessel cancer embolus. CONCLUSIONS: A new predictive panel including PAGE4 expression may help predict liver metastasis of colorectal cancer. PMID- 20010348 TI - Ligation of the intersphincteric fistula tract (LIFT): a sphincter-saving technique for fistula-in-ano. AB - PURPOSE: This study was designed to assess the total anal sphincter-saving technique of ligating the intersphincteric fistula tract for the treatment of fistula-in-ano. METHODS: We performed a prospective observational study of patients with fistula-in-ano treated with the ligation of the intersphincteric fistula tract technique from May 2007 through September 2008. All patients had fistulas arising from cryptoglandular infections. Recurrence rate, healing time, and morbidities related to the procedure were determined with a standard follow up protocol. RESULTS: Forty-five patients with transsphincteric (n = 33) or complex (n = 12) fistulas were included in the study. Five patients (11.1%) had recurrent fistula-in-ano after prior surgery using other recognized treatment procedures. The median age was 41.5 (range, 27-56) years; median follow-up, 9 (range, 2-16) months. Primary healing was achieved in 37 patients (82.2%), with a median healing time of 7 (range, 4-10) weeks. Eight patients (17.7%) had recurrence of fistula between 3 and 8 months after the operation. No clinically significant morbidity was noted in any of the 45 patients. CONCLUSIONS: The ligation of the intersphincteric fistula tract technique for fistula-in-ano surgery, which aims at total anal sphincter preservation, appears to be both safe and easy to perform, with encouraging early outcomes. PMID- 20010349 TI - Ligation of the intersphincteric fistula tract: an effective new technique for complex fistulas. AB - INTRODUCTION: The management of complex fistulas is difficult. Maintaining continence while achieving durable fistula closure is the goal of surgical management. This study describes our experience with a novel sphincter-sparing technique called the ligation of the intersphincteric fistula tract, which involves ligation and division of the fistula tract in the intersphincteric space. METHODS: All patients from July 2007 to December 2008 with trans- or suprasphincteric fistula treated with the procedure were prospectively followed. Procedures were performed by surgeons with fellowship training in a referral center. Demographic data, comorbidities, previous repair attempts, and postoperative data were collected. RESULTS: A total of 39 patients underwent a ligation of the intersphincteric fistula tract during a 17-month period. Median age was 49 years. A total of 29 patients (74%) had previous attempts at repair, with a median of 2 failed repairs. Follow-up data were available in 90% (35 of 39). Median follow-up was 20 weeks. Successful fistula closure was achieved in 57% of the patients (20 of 35). Median time to failure was 10 weeks (range, 2-38 weeks). No patient reported any subjective decrease in continence after the procedure. CONCLUSION: Ligation of the intersphincteric fistula tract is a new sphincter-sparing procedure for complex transsphincteric fistula. The success rate is comparable with other sphincter-preserving techniques. Importantly, it appeared to effectively preserve continence. Adding safe, muscle-sparing surgical options to our armamentarium for dealing with transsphincteric fistula is essential. Additionally, the procedure is easy to learn and has very low cost. Long-term follow-up and randomized, controlled trials are necessary to assess efficacy and durability. PMID- 20010350 TI - Prospective analysis of clinician accuracy in the diagnosis of benign anal pathology: comparison across specialties and years of experience. AB - PURPOSE: The majority of patients referred to a colorectal surgeon with anal complaints are told they have "hemorrhoids"; however, many of these patients have other anal pathology causing their symptoms. Therefore, we prospectively evaluated the diagnostic accuracy of physicians for common anal pathology, stratified by specialty and experience. METHODS: Seven common benign anal pathologic conditions were selected (prolapsed internal hemorrhoid, thrombosed external hemorrhoid, abscess, fissure, fistula, condyloma acuminata, and full thickness rectal prolapse). Prospectively accrued subjects included attending physicians, fellows, residents, and medical students. Subjects were shown images and asked to provide a written diagnosis. We prospectively evaluated the overall diagnostic accuracy and stratified accuracy across specialties and years of clinical experience. Medical students were the control group. RESULTS: There were 198 physicians and 216 medical students. Overall diagnostic accuracy for physicians was 53.5% and for controls was 21.9% (P < .001). Surgeons had the highest overall accuracy at 70.4%, whereas all of the other groups had an accuracy of <50%. Physicians correctly identified condylomata and rectal prolapse most frequently and hemorrhoidal conditions least frequently. All 7 conditions were correctly identified by 4.1% of subjects and all of the conditions were incorrectly diagnosed by 20.2%. There was no correlation between years of experience and diagnostic accuracy (P = NS). CONCLUSION: Diagnostic accuracy for common benign anal pathologic conditions was suboptimal across all clinical specialties. Although many specialties had a diagnostic accuracy that was significantly better than the control group, there was no association between years of experience and accuracy. Improved programs for physician education for these common conditions should be developed. PMID- 20010351 TI - Patients with low rectal cancer treated by abdominoperineal excision have worse tumors and higher involved margin rates compared with patients treated by anterior resection. AB - PURPOSE: Patients with low rectal cancer have worse outcomes compared to those with upper rectal cancer. Reports suggest that low anterior resection may be oncologically superior to abdominoperineal excision, although no good evidence exists to support this. We looked at a recent series of patients with low rectal cancer to explore some of the issues. METHODS: We analyzed 153 patients from the MERCURY study with low rectal cancer (35 kg/m2, diabetes, prostate hypertrophy, ascites, or chronic constipation. CONCLUSION: This study of carefully and prospectively collected data shows the prevalence of parastomal herniation to be 33%. This rate was higher with larger aperture size and increased age in multivariate analysis. PMID- 20010355 TI - Comparison of extended colectomy and limited resection in patients with Lynch syndrome. AB - PURPOSE: The purpose of the study was to determine the advantages and disadvantages of prophylactic/extended colectomy (subtotal colectomy) in patients with Lynch syndrome who manifest colorectal cancer. METHODS: A retrospective cohort using Creighton University's hereditary cancer database was used to identify cases and controls. Cases are patients who underwent subtotal colectomy, either with no colorectal cancer diagnosis (prophylactic) or at diagnosis of first colorectal cancer; controls for these 2 types of cases were, respectively, patients who underwent no colon surgery or those having limited resection at time of diagnosis of first colorectal cancer. The Kaplan-Meier and proportional hazard regression models from the Statistical Analysis Software program was used to calculate the difference in survival, time to subsequent colorectal cancer, and subsequent abdominal surgery between cases and controls. RESULTS: The event-free survival of our study did not reach 50%, so we used the event-free survival at 5 years as our parameter to compare the 2 groups. The event-free survival for subsequent colorectal cancer, subsequent abdominal surgery, and death was 94%, 84%, and 93%, respectively, for cases and 74%, 63%, and 88%, respectively, for controls. Times to subsequent colorectal cancer and subsequent abdominal surgery were significantly shorter in the control group (P < .006 and P < .04, respectively). No significant difference was identified with respect to survival time between the cases and controls. CONCLUSIONS: Even though no survival benefit was identified between the cases and controls the increased incidence of metachronous colorectal cancer and increased abdominal surgeries among controls warrant the recommendation of subtotal colectomy in patients with Lynch syndrome. PMID- 20010356 TI - Outcomes after transanal repair of rectoceles. AB - PURPOSE: To determine the outcomes of patients after transanal rectocele repair. METHODS: The Birmingham Bowel and Urinary Symptoms Questionnaire (BBUSQ-22), a validated instrument to evaluate bowel and urinary symptoms, was completed preoperatively by all patients undergoing transanal rectocele repair and postoperatively at a median interval of 8 months. The BBUSQ-22 was also administered to a control group of 50 asymptomatic female patients. The preoperative and postoperative BBUSQ-22 results for the 9 items pertaining to bowel function were compared to each other and to the responses from the control group. RESULTS: Between April 1, 2001 and December 31, 2003, 88 women underwent transanal rectocele repair. Compared to the control group, patients with rectocele were significantly more symptomatic on all of the questions except the ability to hold bowel movements longer than 5 minutes. A significant improvement was reported postoperatively in all areas except pain with bowel movement and ability to hold bowel movements longer than 5 minutes. When the postoperative responses were compared to the control group, there were no significant differences except for a more frequent need for digital assistance and painful defecation in the surgical group. CONCLUSION: Transanal rectocele repair results in significant improvement in defecation and continence, with postoperative bowel function comparable to control patients in 7 of the 9 areas evaluated. PMID- 20010357 TI - Transanal rectocele repair using linear stapler and bioabsorbable staple line reinforcement material: short-term results of a prospective study. AB - PURPOSE: This study was designed to determine the short-term results of transanal rectocele repair with use of a linear stapler and Bioabsorbable Seamguard. METHODS: Ten women (median age, 56.1 y) with obstructed defecation syndrome were enrolled in the study. The preoperative study consisted of a medical history, physical examination, anoscopy, endoanal ultrasound, and defecography. All patients completed a severity score and a visual analog scale for global quality of life, and provided the postoperative dates of complications. RESULTS: Overall, the treatment significantly improved the obstructed defecation: the severity score improved from a median of 19.8 +/- 4.2 at baseline to 6.10 +/- 8.2 at one month (P < .005), 5.9 +/- 8.3 at 6 months (P < .005), and 6 +/- 8.3 at one year after the operation (P < .005). A significant improvement was observed in the visual analog scale, which improved from a median of 1.8 +/- 1.4 at baseline to 6 +/- 1.6 at one month (P < .005), 6.6 +/- 2.1 at 6 months (P < .004), and 7 +/- 2.5 at one year (P < .004). A significant improvement was also observed in various symptoms. Only 2 patients manifested urgent defecation. CONCLUSIONS: The present study demonstrates that rectocele repair using a linear stapler and Seamguard is a safe, easy procedure, with a very low rate of complications and good immediate outcome. PMID- 20010358 TI - Reduction of oxygenation and blood flow in pedicled bowel segments in the rat and its consequences for anastomotic healing. AB - PURPOSE: Experimental studies indicate that perioperative hypoperfusion impairs anastomotic healing. In bowel surgery, the part of bowel that will be anastomosed is often pedicled, leaving the blood supply dependent on the marginal artery only. Little is known about the blood supply in such a segment, and whether anastomotic strength is affected when flow would be reduced. This study describes oxygenation and blood flow in pedicled bowel segments in the rat and investigates whether early anastomotic strength changes with variations in blood flow. METHODS: In rats, pedicled segments were created in ileum and colon by successive ligation of the feeding arteries. Oxygenation and blood flow were measured in the distal part of this segment by use of near-infrared spectroscopy with indocyanine green as an intravascular tracer. In a second experiment, a short pedicled colonic segment was created and, after flow measurements, an anastomosis was constructed. Wound strength and hydroxyproline content were analyzed 2 and 5 days after operation. RESULTS: After creation of a pedicled segment, the concentration of oxygenated hemoglobin decreased significantly. Blood flow also significantly decreased to even less than 10% of baseline. A very large variation was observed between animals, in particular, after ligation of the first arteries. The strength of colonic anastomoses was not significantly correlated with the blood flow in the pedicled segment before anastomotic construction. CONCLUSIONS: The creation of a pedicled bowel segment greatly reduces tissue oxygenation and blood flow to its distal part. Such impaired perioperative flow does not significantly affect early wound strength after anastomotic construction. PMID- 20010359 TI - Laparoscopic excision of retrorectal schwannoma. AB - The short-term benefits of laparoscopic resection for standard tumors of the colon and rectum are well accepted. Retrorectal tumors are relatively uncommon, and there have been few reports of laparoscopic techniques being applied to such tumors. This video article presents a laparoscopic approach to a retrorectal schwannoma, highlights the technical challenges of such a procedure, and offers some operative tips to aid management of such tumors (see Video, Supplemental Digital Content 1, http://links.lww.com/DCR/A29). PMID- 20010361 TI - Genotype to phenotype-discovery and characterization of novel genomic disorders in a "genotype-first" era. AB - Recent advances in technology for detecting copy number changes have enabled genome-wide detection of submicroscopic deletions and duplications. As these technologies are applied to large cohorts of patients with diverse phenotypes, novel genomic disorders are defined by a common genotype (deletion or duplication) rather than a common phenotype. The discovery of new genomic disorders using this "genotype-first" approach has increased dramatically, and several recently described recurrent rearrangements are associated with a surprisingly wide range of phenotypes. This review will discuss the importance of genomic architecture for generating recurrent rearrangements, implications of the genotype-first approach for medical genetics, and features of several new genomic disorders with highly variable phenotypes. PMID- 20010362 TI - Capturing all disease-causing mutations for clinical and research use: toward an effortless system for the Human Variome Project. AB - The collection of genetic variants that cause inherited disease (causative mutation) has occurred for decades albeit in an ad hoc way, for research and clinical purposes. More recently, the access to collections of mutations causing specific diseases has become essential for appropriate genetic health care. Because information has accumulated, it has become apparent that there are many gaps in our ability to correctly annotate all the changes that are being identified at ever increasing rates. The Human Variome Project (www.humanvariomeproject.org) was initiated to facilitate integrated and systematic collection and access to this data. This manuscript discusses how collection of such data may be facilitated through new software and strategies in the clinical genetics and diagnostic laboratory communities. PMID- 20010363 TI - Lessons from arrhythmogenic right ventricular cardiomyopathy research. PMID- 20010364 TI - Translation of research discoveries to clinical care in arrhythmogenic right ventricular cardiomyopathy in Newfoundland and Labrador: lessons for health policy in genetic disease. AB - Arrhythmogenic right ventricular cardiomyopathy, a lethal autosomal dominant cause of sudden cardiac death in young people, is prevalent in Newfoundland and Labrador (genetic subtype ARVD5). In the absence of implantable cardioverter defibrillator treatment, death rates are extremely high. Research into arrhythmogenic right ventricular cardiomyopathy (ARVD5) began in the 1980s and the causative gene and mutation were discovered in 2008. The decades of research highlighted major issues associated with the ethical management of genetic information and the translation of research findings to clinical care. We describe these issues and the strategies used in managing them. Effective knowledge transfer of the research information has resulted in systematic clinical and genetic screening coupled with genetic counseling and treatment for at-risk family members. Improved survival for patients has been one clear result of this strategy. Optimal care of families where individuals are at-high risk of inheriting a disease with high morbidity and mortality requires the full integration of both genetic research and clinical genetics programs. Although yet to be fully effected in our setting, our discussion highlights both the ethical necessity as well as some practical barriers in realizing this outcome. PMID- 20010365 TI - "Am I my genes?": Questions of identity among individuals confronting genetic disease. AB - PURPOSE: : To explore many questions raised by genetics concerning personal identities that have not been fully investigated. METHODS: : We interviewed in depth, for 2 hours each, 64 individuals who had or were at risk for Huntington disease, breast cancer, or alpha-1 antitrypsin deficiency. RESULTS: : These individuals struggled with several difficult issues of identity. They drew on a range of genotypes and phenotypes (e.g., family history alone; mutations, but no symptoms; or symptoms). They often felt that their predicament did not fit preexisting categories well (e.g., "sick," "healthy," "disabled," "predisposed"), due in part to uncertainties involved (e.g., unclear prognoses, since mutations may not produce symptoms). Hence, individuals varied in how much genetics affected their identity, in what ways, and how negatively. Factors emerged related to disease, family history, and other sources of identity. These identities may, in turn, shape disclosure, coping, and other health decisions. CONCLUSIONS: : Individuals struggle to construct a genetic identity. They view genetic information in highly subjective ways, varying widely in what aspects of genetic information they focus on and how. These data have important implications for education of providers (to assist patients with these issues), patients, and family members; and for research, to understand these issues more fully. PMID- 20010366 TI - Section E6.5 of the ACMG technical standards and guidelines: chromosome studies for solid tumor abnormalities. AB - PURPOSE: : Cytogenetic analysis of tumor tissue detects clonal abnormalities. The information obtained from these studies is utilized for diagnosis, prognosis, and patient management. METHODS: : The Working Group of the Laboratory Quality Assurance Committee of the American College of Medical Genetics provides these Standards and Guidelines for chromosome studies for solid tumors abnormalities as a resource for clinical cytogenetic laboratories. RESULTS: : The guidelines incorporate aspects of sample procurement, handling, processing, harvesting, analysis, quality control, and quality assurance. It is recommended that all pediatric solid tumors be studied by cytogenetic analysis when feasible due to the clinical and therapeutic implications of the genetic abnormalities. Cytogenetic analysis of certain adult solid tumors also provides information that impacts diagnosis and therapeutics. Molecular cytogenetic analysis or fluorescence in situ hybridization (FISH) may be a primary or secondary method of evaluation of the tumor tissue. FISH can document a specific molecular event, e.g. gene rearrangement, provide a rapid result to aid in the differential diagnosis or planning of therapy, clarify chromosome anomalies, or assess gene amplification. CONCLUSION: : Genetic analysis adds valuable information to the understanding of and therapeutic approach to solid tumors. Laboratories may use their professional judgment to make modifications or additions to these guidelines. PMID- 20010367 TI - Realizing the potential of genomics: translation is not translational research. PMID- 20010369 TI - Pandemic influenza: allocating scarce critical care resources. PMID- 20010370 TI - Tips for writing magnet evidence. PMID- 20010371 TI - Creating quality evidence summaries on a clinician's schedule. PMID- 20010372 TI - Assessing and selecting data for a nursing services dashboard. AB - Nurse leaders are held accountable for staffing decisions and quality outcomes, sometimes without proper decision support tools. A nursing services dashboard can provide information to guide staffing decisions for positive outcomes. The authors examine the sources, attributes, and uses of administrative data to assist nurse leaders to make informed decisions in selecting data for inclusion in their nursing services dashboard. PMID- 20010373 TI - The nurse-nurse collaboration scale. AB - OBJECTIVE: To develop and examine the reliability and validity of a new instrument, the nurse-nurse collaboration (NNC) scale. BACKGROUND: Nurse-nurse collaboration (NNC) is recommended to reduce medical errors and improve patient care and nurses' job satisfaction. While instruments are available to measure nurse-physician collaboration, an instrument to measure NNC was not available in the literature. Because collaboration is necessary for optimal patient care, a valid and reliable instrument would make it possible to measure the level of collaboration among nurses. METHODS: A comprehensive literature review was conducted to develop a definition and define relevant domains of NNC and identify instruments with acceptable psychometrics that included items measuring NNC. Instrument items to develop the Nurse-Nurse Collaboration Scale (NNC Scale) were adapted from previously published tools or developed based on domains identified in the literature. Five domains were identified: problem solving, communication, coordination, shared process, and professionalism. Psychometric testing of the NNC Scale included pilot testing for content and construct validity and field testing among 76 staff nurses working in 4 ICUs in a large tertiary-care academic medical center in the northeast United States. Psychometric tests assessing reliability and convergent validity correlations were conducted. RESULTS: The overall Cronbach alpha for the scale was .89. Convergent validity correlations, however, were low to moderate, indicating minimal shared variance among the subscales. Therefore, the instrument did not measure a global concept but rather 5 separate domains of collaboration. Internal consistency testing of the 5 subscales produced acceptable results ranging from .66 to .91. CONCLUSION: The NNC Scale demonstrated acceptable reliability and validity for measuring the level of NNC in intensive care nurses. Further psychometric testing and a factor analysis with a larger-sample, more diverse groups of nurses are necessary to further characterize the generalizability of the NNC Scale. PMID- 20010374 TI - Matching patient safety goals to the nursing specialty: using wound, ostomy, continence nursing services. AB - With increasing nursing specialization, administrators may not have clear guidelines directing which specialists should be targeting specific patient populations and specific patient care issues. Because pressure ulcers are an important indicator of quality of care, this discussion focuses on selecting the appropriate wound, ostomy, continence specialist to develop and support programs that are designed to prevent pressure ulcers, treat patients with pressure ulcers, as well as management of other types of wounds, stomas, fistulas, incontinence, and skin breakdown. Nurse leaders are in a position to ensure that appropriate resources are available to support the development of hospital programs that will promote safe, efficient, and cost-conscious patient care. PMID- 20010375 TI - Job embeddedness as a nurse retention strategy for rural hospitals. AB - RN turnover is expensive and disruptive for rural hospitals, constraining finances, impacting patient care, and stressing remaining nurses. Recent investigations have described a promising new construct related to employee retention: job embeddedness. Leaders in nonhealthcare organizations have adopted a job-embeddedness model to guide retention strategies and experienced a subsequent reduction in turnover. The author explores job embeddedness as an effective retention plan strategy for rural hospitals. PMID- 20010376 TI - Assessing and redesigning the nursing practice environment. AB - With an aging population, aging nurse workforce, and high nurse vacancy rates, our hospital had a clear mandate to assess and redesign our practice environment. The authors describe a project created to provide a framework for current and ongoing evaluation of the practice environment. PMID- 20010377 TI - Nursing research and participant recruitment: organizational challenges and strategies. AB - Hospitals as research environments are crucial in advancing evidence-based practice and translational research. The authors discuss issues related to hospital-based nursing research such as institutional review board approval, the HIPAA, structure, unit characteristics, and nurse staffing as well as research related issues such as study purpose and design, participant recruitment, and research personnel. Strategies and suggestions for nurse executives to assist researchers in overcoming recruitment challenges are presented. PMID- 20010379 TI - Lung volume measurement in the neonate-throwing light on the subject: commentary on the article by Dellaca' et al. on page 11. PMID- 20010380 TI - Introduction of the American Pediatric Society's 2009 John Howland Award Recipient, Jerold F. Lucey, M.D. PMID- 20010381 TI - American Pediatric Society's 2009 John Howland award acceptance lecture: lessons learned from time. PMID- 20010382 TI - Polymorphic variants in the human bile salt export pump (BSEP; ABCB11): functional characterization and interindividual variability. AB - OBJECTIVES: Our aims were to identify and functionally characterize coding region nonsynonymous single nucleotide polymorphisms in the hepatic efflux transporter, bile salt export pump (BSEP; ABCB11), and to assess interindividual variability in BSEP expression. METHODS: We identified 24 single nucleotide polymorphisms, including nine nonsynonymous variants, in ABCB11 from genomic DNA of approximately 250 ethnically diverse healthy individuals using denaturing high performance liquid chromatography analysis and DNA sequencing. Wild type and variant BSEP were generated and functionally characterized for taurocholate transport activity in vitro in HeLa cells using a recombinant vaccinia-based method. BSEP expression was assessed by real-time mRNA analysis, western blot analysis, and immunofluorescence confocal microscopy. RESULTS: For the most part, polymorphisms were rare and ethnic-dependent. In vitro functional studies revealed several rare variants, including 616A>G, 1674G>C, 1772A>G, and 3556G>A, to be associated with significantly impaired taurocholate transport activity while the 890A>G variant trended towards impaired function but was not statistically significant. The 3556G>A variant was associated with reduced cell surface to total protein expression compared with wild-type BSEP. Expression of BSEP by mRNA and protein analysis was determined from a bank of human liver samples. Wide interindividual variability was noted in both mRNA (19-fold) and protein (31-fold) expression levels. The common variant 1331T>C was associated with significantly reduced hepatic BSEP mRNA levels. CONCLUSION: Accordingly, our study indicates there are functionally relevant polymorphisms in ABCB11 which may be of potential relevance in the predisposition to acquired liver disorders such as drug-induced cholestasis. PMID- 20010383 TI - Lidocaine cytotoxicity to the zygapophysial joints in rabbits: changes in cell viability and proteoglycan metabolism in vitro. AB - STUDY DESIGN.: To examine whether lidocaine cytotoxicity to chondrocytes has been implicated in the development of osteoarthritis of the zygapophysial joints. OBJECTIVE.: This study was performed to determine the effects of varying concentrations and exposure times of lidocaine on the viability and proteoglycan metabolism of rabbit zygapophysial chondrocytes in vitro. SUMMARY OF BACKGROUND DATA.: Zygapophysial joint injections are commonly administered with lidocaine for chronic spinal pain in orthopedic treatment. A lot of studies on the effect of zygapophysial joint injections are clinical, but many questions on the effect of lidocaine to zygapophysial chondrocytes remain unanswered. METHODS.: Cartilage was obtained from zygapophysial joints of adult rabbits. Chondrocytes in alginate beads were cultured in medium containing 6% fetal calf serum at 370 mOsmol at cell densities of 4 million cells/mL. They were then cultured for 24 hours under 21% oxygen with 0.125%, 0.25%, 0.5%, and 1% lidocaine, and without lidocaine as control. The cell viability profile across intact beads was determined by manual counting using fluorescent probes (LIVE/DEAD assay) and transmission electron microscopy. Lactate production was measured enzymatically as a marker of energy metabolism. Glycosaminoglycan (GAG) accumulation was measured using a modified dimethylmethylene blue assay. RESULTS.: Cell viability decreased in a time- and dose-dependent manner in the concentration range of 0.125% to 1.0% lidocaine under the confocal microscope. Under the electron microscope, apoptosis increased as the concentration of lidocaine increased. GAG accumulation/tissue volume decreases as the concentration of lidocaine increased. However, GAG produced per million cells and the rate of lactate production per live cell was significantly higher for cells cultured at 0.5% and 1% lidocaine than the control group. CONCLUSION.: While these in vitro results cannot be directly extrapolated to the clinical setting, this data suggestcaution in prolonged exposure of zygapophysial cartilage to high concentration lidocaine. PMID- 20010384 TI - Role of the oxygen-dependent degradation domain in a hypoxia-inducible gene expression system in vascular endothelial growth factor gene therapy. AB - STUDY DESIGN.: An in vitro neural hypoxia model and rat spinal cord injury (SCI) model were used to assess the regulation effect of a reporter or therapeutic gene expression by an oxygen-dependent degradation (ODD) domain in a hypoxia-inducible gene expression system with or without the erythropoietin (EPO) enhancer. OBJECTIVE.: To increase vascular endothelial growth factor (VEGF) gene expression in SCI lesions but avoid unwanted overexpression of VEGF in normal sites, we developed a hypoxia-inducible gene expression system consisting of the EPO enhancer upstream of the SV promoter and an ODD domain C-terminally fused to VEGF. SUMMARY OF BACKGROUND DATA.: ODD domain plays a major role in the degradation of hypoxia-inducible factor 1alpha and has been used in a hypoxia specific gene expression system as a post-translational regulatory factor. METHODS.: The hypoxia-inducible luciferase or VEGF plasmid was constructed using the EPO enhancer combined with or without the ODD domain. The constructed plasmid was transfected into mouse Neuro 2a (N2a) neuroblastoma cells by Lipofectamine 2000, followed by a 24-hour incubation in hypoxia or normoxia. For in vivo analysis, the naked plasmid DNA was directly injected into the injured rat spinal cord. The gene expression was evaluated by luciferase activity assay, enzyme linked immunosorbent assay, reverse transcriptase-polymerase chain reaction, and immunofluorescence staining. RESULTS.: The EPO enhancer/ODD domain-combined hypoxia-inducible gene expression system clearly increased the expression of the reporter luciferase gene and therapeutic VEGF gene specifically under hypoxic conditions and SCI, and quickly downregulated protein expression to a very low level after reoxygenation. CONCLUSION.: These results strongly suggest the potential applicability of this EPO enhancer/ODD domain-based hypoxia-inducible gene expression system in the development of a safer and more effective VEGF gene therapy for SCI. PMID- 20010385 TI - A novel patient-specific navigational template for cervical pedicle screw placement. AB - STUDY DESIGN.: Prospective trial. OBJECTIVE.: To develop and validate a novel, patient-specific navigational template for cervical pedicle placement. SUMMARY OF BACKGROUND DATA.: Owing to the narrow bony anatomy and the proximity to the vertebral artery and the spinal cord, cervical instrumentation procedures demand the need for a precise technique for screw placement. PATIENT.: Specific drill template with preplanned trajectory has been thought as a promising solution for cervical pedicle screw placement. METHODS.: Patients with cervical spinal pathology (n = 25) requiring instrumentation were recruited. Volumetric CT scan was performed on each desired cervical vertebra and a 3-dimensional reconstruction model was generated from the scan data. Using reverse engineering technique, the optimal screw size and orientation were determined and a drill template was designed with a surface that is the inverse of the posterior vertebral surface. The drill template and its corresponding vertebra were manufactured using rapid prototyping technique and tested for violations. The navigational template was sterilized and used intraoperatively to assist with the placement of cervical screws. In total, 88 screws were inserted into levels C2-C7 with 2 to 6 screw in each patient. After surgery, the positions of the pedicle screws were evaluated using CT scan and graded for validation. RESULTS.: This method showed its ability to customize the placement and the size of each screw based on the unique morphology of the cervical vertebra. In all the cases, it was relatively very easy to manually place the drill template on the lamina of the vertebral body during the surgery. The required time between fixation of the template to the lamina and insertion of the pedicle screws was about 80 seconds. Of the 88 screws, 71 screws had no deviation and 14 screws had deviation <2 mm, 1 screw had a deviation between 2 to 4 mm and there were no misplacements. Fluoroscopy was used only once for every patient after the insertion of all the pedicle screws. CONCLUSION.: The authors have developed a novel patient-specific navigational template for cervical pedicle screw placement with good applicability and high accuracy. This method significantly reduces the operation time and radiation exposure for the members of the surgical team. The potential use of such a navigational template to insert cervical pedicle screws is promising. This technique has been clinically validated to provide an accurate trajectory for pedicle screw placement in the cervical spine. PMID- 20010386 TI - Progressive kyphoscoliosis associated with tethered cord treated by posterior vertebral column resection: a case report. AB - STUDY DESIGN.: A case report. OBJECTIVES.: To report a case of progressive kyphoscoliosis associated with a tethered cord that was corrected by posterior vertebral column resection after complicated untethering surgery. SUMMARY OF BACKGROUND DATA.: There have been few clinical reports on posterior vertebral column resection conducted for severe deformity associated with a tethered cord. METHODS.: A patient with progressive kyphoscoliosis associated with a tethered cord first underwent untethering surgery, resulting in neurologic deterioration. Posterior vertebral column resection was performed to correct the kyphoscoliosis while shortening the spinal column to prevent the spinal cord from stretch injury. RESULTS.: Good correction of kyphoscoliosis was obtained without further neurologic deterioration. The Cobb angles of scoliosis was 103 degrees before surgery and 25 degrees after surgery (correction rate; 75.7%), and that of kyphosis was 90 degrees and 36 degrees , respectively (correction rate; 60.0%). CONCLUSION.: Correction of progressive kyphoscoliosis associated with a tethered cord can be achieved successfully by posterior vertebral column resection even after complicated untethering surgery. PMID- 20010387 TI - A rare case of extramedullary-intradural hemangioblastoma in the thoracic spine. AB - STUDY DESIGN.: Case report. OBJECTIVE.: To present a rare case of extramedullary intradural hemangioblastoma in the thoracic spine and to review the literature on this condition. SUMMARY OF BACKGROUND DATA.: Spinal hemangioblastoma is rare, and moreover, there is seldom purely extramedullary-intradural hemangioblastoma of the spinal cord and there are few reports on histopathological features of spinal hemangioblastoma. METHODS.: A 65-year-old Japanese woman presented with gait disturbance and numbness below the trunk that had gradually worsened, and finally she was unable to walk by herself. Physiologic and neurologic examinations at admission revealed severe transverse neurologic defects, with no sign of von Hippel-Lindau's disease. Radiologic methods showed an extramedullary-intradural mass at Th4-5. A diagnosis of extramedullary-intradural spinal tumor was done, total tumorectomy was performed with recapped laminoplasty. RESULTS.: The patient's neurologic condition gradually improved after the operation. The immunohistopathological findings revealed the diagnosis of hemangioblastoma. CONCLUSION.: A rare case of extramedullary-intradural spinal hemangioblastoma was reported. The immunohistopathological findings were helpful for making a final diagnosis. PMID- 20010388 TI - The rare case of an intramedullary cervical spinal cord teratoma in an elderly adult: case report and literature review. AB - STUDY DESIGN.: Case report and literature review. OBJECTIVES.: To report the very rare case of a mature intramedullary teratoma with exophytic extension localized to the uppermost cervical spinal level in a 65-year-old woman and review the pertinent medical literature. SUMMARY OF BACKGROUND DATA.: Cervical intramedullary teratomas are extremely rare in adults, especially in patients older than 50 years. METHODS.: The patient presented with progressive ataxia, mild bilateral kinetic hand tremors, and dizziness. Magnetic resonance imaging revealed an intramedullary 1.7 x 1.3 x 2.3 cm mass at C1 with exophytic extension. A C1-C2 laminectomy and a partial suboccipital craniotomy were performed, followed by a subtotal microscopic resection of the tumor. Pathology was consistent with a mature teratoma. RESULTS.: After surgery, the patient's ataxia, tremor, and dizziness resolved almost immediately. CONCLUSION.: This report presents the very rare case of a mature intramedullary teratoma located in the upper cervical spine of an elderly patient, possibly the oldest patient documented with this type of lesion. The authors recommend a conservative subtotal surgical resection of cervical intramedullary tumors because it may improve symptoms that relate to direct mechanical cord compression and avoid further harm from a gross resection. PMID- 20010389 TI - Symptomatic epidural pneumorrhachis associated with an occult pneumomediastinum due to minor trauma. AB - STUDY DESIGN.: A case report and review of the literature. OBJECTIVE.: To discuss the significance of identifying symptomatic epidural pneumorrhachis associated with an occult pneumomediastinum in the absence of pneumothorax and subcutaneous emphysema after minor trauma. SUMMARY OF BACKGROUND DATA.: Pneumorrhachis is defined as the presence of air in the epidural space or subarachnoid space. Previous reports have suggested that traumatic epidural pneumorrhachis was usually asymptomatic and a combination of pneumothorax, pneumomediastinum, and subcutaneous emphysema following major trauma and managed by conservative treatment. METHODS.: A 53-year-old man was admitted to the emergency department after the traffic collision. He presented with diffuse pain ranging from the neck to back, and complained of radiating shoulder pain. RESULTS.: The lateral cervical spine radiograph revealed an air shadow in the anterior cervical space. Axial image of cervical spine computed tomography (CT) revealed epidural pneumorrhachis. The sagittal reconstruction image of neck CT showed a small amount of air in the posterior mediastinum, and scattered air in the anterior cervical spinal canal. Our patient was treated conservatively and had a complete and uneventful recovery. CONCLUSION.: To our knowledge, this is the first case in which symptomatic traumatic epidural pneumorrhachis was associated with an occult pneumomediastinum in the absence of pneumothorax and subcutaneous emphysema. These findings may be useful in alerting trauma specialists to carefully evaluate the associated pathologies leading to pneumorrhachis enabling adequate therapy. PMID- 20010391 TI - Anomalous vertebral artery in craniovertebral junction with occipitalization of the atlas. AB - STUDY DESIGN.: Observational study with 3-dimensional computed tomography angiography analysis. OBJECTIVE.: To examine the course of the vertebral artery (VA) at the craniovertebral junction (CVJ) in individuals with occipitalization of the atlas. SUMMARY OF BACKGROUND DATA.: The anatomy of the VA at the CVJ should be completely understood to decrease the risk of iatrogenic injury. Although quantitative anatomic studies have focused on the normal VA, the anomalous VA with occipitalization of the atlas has not been fully explored. METHODS.: A consecutive series of 36 cases with occipitalization of the atlas underwent 3-dimensional computed tomography angiography. Seventy-two vertebral arteries were analyzed. In this setting, the safety of placing lateral mass screws (LMS) was studied. RESULTS.: Four different pathways of the VA at the CVJ with occipitalization of the atlas were found. Type I, wherein the VA enters the spinal canal below the C1 posterior arch, and the course of the VA is below the occipitalized C1 lateral mass (8.3% of 72 vertebral arteries); Type II, the VA enters the spinal canal below the C1 posterior arch, and the course of the VA is on the posterior surface of the occipitalized C1 lateral mass, or makes a curve on it (25%); Type III, wherein the VA ascends externally laterally after leaving the axis transverse foramen, enters an osseous foramen created between the atlas and occipital bone, then into the cranium (61.1%); and Type IV, in which the VA is absent (5.6%). CONCLUSION.: Four types of VA with occipitalization of the atlas are confirmed. Type-I and type-IV VA have relatively low risks for C1 LMS perforation. Type-II and type-III anomalies will probably increase the risk of VA injury during C1 LMS placement. Definite caution should also be taken during the procedure on the contralateral side of a type-IV VA. PMID- 20010392 TI - Direct single injection of p38 mitogen-activated protein kinase inhibitor does not affect calcitonin gene-related peptide expression in dorsal root ganglion neurons innervating punctured discs in rats. AB - STUDY DESIGN.: Immunohistological analysis of punctured disc after application of a p38 MAP kinase inhibitor. OBJECTIVE.: To examine effect of direct application on dorsal root ganglion (DRG) neurons innervating damaged rat discs. SUMMARY OF BACKGROUND DATA.: Degeneration of lumbar discs is one cause of low back pain. Pathogenesis may involve sensory nerve ingrowth into disc inner layers; tumor necrosis factor-alpha (TNF-alpha) is thought to be a major inducer of ingrowth. Because p38 mitogen-activated protein kinase (p38) upregulates TNF-alpha expression and may play a crucial role in pain sensation, we investigated the effect of one injection of inhibitor on expression of the pain-related neuropeptide calcitonin gene-related peptide (CGRP). METHODS.: The neuro-tracer fluoro-gold was applied to the surfaces of L4/5 discs to label the innervating DRG neurons (n = 30). Of 30 rats, 10 were controls, whereas the other 20 were the experimental model (i.e., discs were punctured with 23-gauge needle). P38 specific inhibitor or saline was applied simultaneously (n = 10 each, Puncture + inhibitor and puncture + saline groups). Fourteen days postsurgery, DRGs from L1 to L6 were harvested, sectioned, and immunostained for CGRP. Proportion of CGRP immunoreactive DRG neurons was evaluated in all groups. RESULTS.: Fluoro-gold labeled neurons innervating the L4/5 disc were distributed throughout L1 to L6 DRGs in all groups. Proportions of labeled neurons positive for CGRP were 15.2% +/- 8% (controls), 27.2% +/- 10% (puncture + saline), and 25.2% +/- 8% (puncture + inhibitor). Proportion of immunoreactive neurons was significantly increased in the puncture groups compared with controls. However, there was no significant difference between the 2 puncture groups (P > 0.1). CONCLUSION.: In this model, CGRP was upregulated in DRG neurons innervating the damaged disc. However, a direct single application of p38 inhibitor did not suppress CGRP expression in innervating DRG neurons. Future research with p38 inhibitor in this model should evaluate multiple or systemic administration of inhibitor. PMID- 20010393 TI - Biomechanical characteristics of different regions of the human spine: an in vitro study on multilevel spinal segments. AB - STUDY DESIGN.: An in vitro study on human multilevel spinal segments. OBJECTIVE.: To determine the differences in biomechanical characteristics between 4 separate regions of the human spine and to provide quantitative information is derived on the range of motion (ROM), neutral zone (NZ), neutral zone stiffness (NZstiff), and flexibility (FLEX). SUMMARY OF BACKGROUND DATA.: Limited literature is available about the biomechanical behavior of different regions of the human spine, in particular with multilevel segments. Test setup en protocols were different between studies and therefore outcomes of separate regions are hardly comparable. METHODS.: A total of 24 spinal segments of 6 human cadaveric spines were prepared for biomechanical testing. Each specimen contained 4 vertebrae and 3 intervertebral discs: T1-T4, T5-T8, T9-T12, and L1-L4. Pure moments were applied to a maximum of 4 Nm in flexion/extension, lateral bending, and axial rotation. Displacement of individual motion segments was measured using a 3 dimensional movement registration system. ROM, NZ, NZstiff, and FLEX of the spinal regions were calculated from the acquired load-displacement data. RESULTS.: In axial direction, ROM and NZ decreased and NZ stiffness increased from high to low vertebral levels. For flexion/extension and lateral flexion highest ROM and NZ and lowest NZ stiffness values were found at the T1-T4 and L1 L4 regions. NZ magnitudes and NZ stiffnesses were negatively correlated (P < 0.05). Flexibility of the spinal regions was variable; no significant differences were found between the 4 spinal regions. CONCLUSION.: This study showed the differences in ROM, NZ, and NZ stiffness between thoracolumbar regions of the human spine in axial rotation, flexion/extension, and lateral bending. Separate multilevel spinal segments were tested in 1 study, and therefore characteristics of different regions are truly comparable. PMID- 20010394 TI - Vertebroplasty: only small cement volumes are required to normalize stress distributions on the vertebral bodies. AB - STUDY DESIGN.: Biomechanical study of vertebroplasty in cadaver motion segments. OBJECTIVES.: To determine how the volume of injected cement influences: (a) stress distributions on fractured and adjacent vertebral bodies, (b) load-sharing between the vertebral bodies and neural arch, and (c) cement leakage. SUMMARY OF BACKGROUND DATA.: Vertebroplasty is increasingly used to treat vertebral fractures, but there are problems concerning adjacent level fracture and cement leakage, both of which may depend on the volume of injected cement. METHODS.: Nineteen thoracolumbar motion segments from 13 cadavers (42-91 years) were loaded to induce fracture. Fractured vertebrae received 2 sequential injections (VP1 and VP2) of 3.5 cm of polymethylmethacrylate cement. Before and after each intervention, motion segment stiffness was measured in compression and in bending, and "stress profilometry" was used to quantify the distribution of compressive stress in the intervertebral disc (which presses equally on fractured and adjacent vertebrae). Stress profiles were obtained by pulling a pressure transducer through the disc while the motion segment was compressed in flexed and extended postures. Stress profiles yielded the intradiscal pressure (IDP), the magnitude of stress peaks in the anterior and posterior (SPP) anulus, and the percentage of the applied compressive force resisted by the neural arch (FN). Cement leakage and vertebral body volume were quantified using water-immersion, and the percentage cement fill was estimated. RESULTS.: Bending and compressive stiffness fell by 37% and 50% respectively following fracture, and were restored only after VP2. Depending on posture, IDP fell by 59-85% after fracture whereas SPP increased by 107- 362%. VP1 restored IDP and SPP to prefracture values, and VP2 produced no further changes. Fracture increased FN from 11% to 39% in flexion, and from 33% to 59% in extension. FN was restored towards prefracture values only after VP2. Cement leakage increased after VP2 and was negatively correlated to vertebral body volume. Following VP2, increases in IDP and compressive stiffness were proportional to percentage fill. CONCLUSION.: About 3.5 cm of PMMA largely restored normal stress distributions to fractured and adjacent vertebral bodies, but 7 cm were required to restore motion segment stiffness and load-sharing between the vertebral bodies and neural arch. Cement leakage, IDP and compressive stiffness all increased with percentage fill. PMID- 20010395 TI - C1-C2 transarticular screw fixation for atlantoaxial instability due to rheumatoid arthritis: a seven-year analysis of outcome. AB - STUDY DESIGN.: Observational study. Retrospective analysis of prospectively collected data. OBJECTIVE.: The purpose of this article was to report long-term (minimum 7 years) clinical and radiologic outcome of our series of patients with Rheumatoid Arthritis who underwent transarticular screw fixation to treat atlantoaxial subluxation. SUMMARY OF BACKGROUND DATA.: The indications for intervention in patients with atlantoaxial instability are pain, myelopathy, and progressive neurologic deficit. The various treatment options available for these patients are isolated C1-C2 fusion, occipitocervical fusion with or without transoral surgery. Review of current literature suggests that C1-C2 transarticular screw fixation has significant functional benefits, although there is discrepancy in this literature regarding improvement in function following surgery. METHODS.: Myelopathy was assessed using Ranawat myelopathy score and Myelopathy Disability Index. Pain scores were assessed using Visual Analogue Scale. The radiologic imaging was assessed and the following data were extracted; atlanto-dens interval, space available for cord, presence of signal change on T2 weighted image, and fusion rates. RESULTS.: Thirty-seven patients, median age 56, were included in the study. Average duration of neck symptoms was 15.8 months. Average duration of rheumatoid arthritis before surgery was 20.6 years. Preoperative symptoms: suboccipital pain in 26 patients; neck pain, 32; myelopathy, 22; and 5 were asymptomatic. After surgery: suboccipital pain, 2; neck pain, 3; and myelopathy, 10. Ninety percent patients with neck and suboccipital pain improved after surgery in their Visual Analogue pain scores, with all of them having >50% improvement in VAS scores (6.94-2.12 [P < 0.05]).Preoperative Ranawat grade was as follows: grade 1 in 15 patients, grade 2 in 7, and grade 3a in 14, grade 3b in 1.After surgery: grade 1 in 27 patients, grade 2 in 7, grade 3a in 1, and grade 3b in 2. The mean myelopathy score improved after surgery (59.62-32.75, P < 0.05).The space available for the cord was improved in 63%, unchanged in 33%, and worse in 4%.Twenty-seven percent had T2 signal change and 18% had cervicomedullary compression; 97% had bony fusion. BILATERAL SCREWS WERE USED IN 33 PATIENTS AND UNILATERAL SCREWS IN 4 PATIENTS (ABERRANT VERTEBRAL ARTERY).: Computer image guidance was used in 73%. CONCLUSION.: C1-C2 transarticular screw fixation is a safe technique for atlantoaxial subluxation for patients with rheumatoid arthritis. This study clearly demonstrates improvement in Visual Analogue Scale, Ranawat grading and the Myelopathy Disability Index even at long-term follow up. PMID- 20010396 TI - Thoracic pedicle subtraction osteotomy for fixed sagittal spinal deformity. AB - STUDY DESIGN.: A retrospective clinical study. OBJECTIVE.: To find the corrective capacity of a thoracic pedicle subtraction osteotomy (PSO), determine if segmental correction is dependent on level, and to compute the impact of thoracic PSO on regional and global spinal balance. SUMMARY OF BACKGROUND DATA.: PSO is a technique popularized in the lumbar spine primarily for the correction of fixed sagittal imbalance. Despite several studies describing the clinical and radiographic outcome of lumbar PSO, there is no study in literature reporting its application in the thoracic spine. METHODS.: We retrospectively analyzed patients with fixed thoracic kyphosis who underwent thoracic PSOs for sagittal realignment. Segmental pedicle screw instrumentation and intraoperative neurophysiologic monitoring was used in all patients. Data acquisition was performed by reviewing medical charts and radiographs to determine sagittal correction (segmental/regional/global) and complications. Clinical outcome using the Scoliosis Research Society-22 (SRS-22) instrument was determined by interview. RESULTS.: A total of 25 thoracic PSOs were performed (mean: 1.7 PSOs/patient, range: 1-3) in 15 patients (9 M/6 F). The study population had an average age of 56 years (range, 36-81 years) and was followed up after surgery for a mean of 3.5 years (range, 24-75 months). The osteotomies were carried out in the proximal thoracic spine (T2-T4, n = 6), midthoracic spine (T5-T8, n = 12), and distal thoracic spine (T9-T12, n = 7). Mean correction at the PSO for all 25 levels was 16.3 degrees +/- 9.6 degrees . Stratified by region of the spine, thoracic PSO correction was as follows: T2-T4 = 10.7 degrees +/- 15.8 degrees , T5-T8 = 14.7 degrees +/- 4.6 degrees , and T9-T12 = 23.9 degrees +/- 4.1 degrees . Mean thoracic kyphosis (T2-T12 Cobb angle) was improved from 75.7 degrees +/- 30.9 degrees to 54.3 degrees +/- 21.4 degrees resulting in a significant regional sagittal correction of 21.4 degrees +/- 13.7 degrees (P < 0.005). Global sagittal balance was improved from 106.1 +/- 56.6 to 38.8 +/- 37.0 mm yielding a mean correction of 67.3 +/- 54.7 mm (P < 0.005). One patient, in whom there was segmental translation during osteotomy closure, had a decline in intraoperative somatosensory-evoked potentials. No patient sustained a temporary or permanent neurologic deficit after surgery. The mean SRS-22 Questionnaire score at final follow-up was 82.4 +/- 10.2. CONCLUSION.: Thoracic PSO can be performed safely. Segmental sagittal correction appears to vary based on the region of the thoracic spine the PSO is performed. The distal thoracic segments, which more closely resemble lumbar segments in morphology, rendered the greatest sagittal correction after PSO, approximately 24 degrees . There was no case of neurologic injury associated with thoracic PSO, and clinical outcomes according to the SRS-22 instrument were generally favorable. PMID- 20010397 TI - Three-dimensional analysis of the cervical spine kinematics: effect of age and gender in healthy subjects. AB - STUDY DESIGN.: A three-dimensional (3D) analysis of the cervical spine kinematics in vivo about a large asymptomatic database in order to evaluate the impact of age and gender on the neck's performances. OBJECTIVE.: To investigate the effect of age and gender on kinematical parameters of the cervical spine, specifically quantitative parameters concerning coupled movements and proprioception, using the infra-red POLARIS measurement system. SUMMARY OF BACKGROUND DATA.: Cervical spine kinematics has been investigated in vivo by numerous authors using various devices. However, few is known about the influence of gender and age on the 3D cervical biomechanics, specifically regarding coupled movements and proprioceptive abilities. METHODS.: A total of 140 asymptomatic volunteers (70 men and 70 women) aged 20 to 93 years old were enrolled. The noninvasive infrared system Polaris was used to quantify the 3D range of motion (ROM) of cervical spine and to evaluate proprioceptive abilities. For validating the protocol in terms of reproducibility, 12 volunteers were tested 3 times by 2 independent operators. RESULTS.: The standard error of measurement for the maximal ROM in the 3 space planes was 5%. Gender had no significant influence on the 3D cervical ROM, except for the "70-79 years old" group. Age had a significant influence on all main movements showing 0.55 degrees to 0.79 degrees magnitude decrease per decade. Age and gender had no significant influence on coupled movements. "Head to-Target" proprioception was significantly affected by the age only in the horizontal plane. CONCLUSION.: A data base for cervical ROM, pattern of motion, and proprioceptive capability was established in population of 140 healthy subjects of various age and gender. Significant age-related decrease in ROM and proprioceptive abilities were observed in this study. Coupled movements did not vary with gender or age; however, their role in the cervical performance increased with age since main movements were limited. PMID- 20010398 TI - Feasibility and accuracy of pedicle screws in children younger than eight years of age. AB - STUDY DESIGN.: A retrospective review. OBJECTIVE.: To determine the feasibility and accuracy of pedicle screw placement in children younger than 8 years of age. SUMMARY OF BACKGROUND DATA.: Computed tomography (CT) scanning is commonly used for assessing the breach associated with pedicle screw placement. The accuracy of pedicle screw placement as evaluated by CT has been described for adults and older children. No study has been published describing pedicle screw breach rate as evaluated by CT scanning in children younger than 8 years of age. METHODS.: A retrospective review of 16 children, with an average age of 4 years 7 months (range, 2 years 3 months to 7 years 11 months) with insertion of 88 pedicle screws was performed. These patients underwent postoperative CT scan. Breach rate was assessed using previously described criteria by Kim et al in 2005. RESULTS.: No patient experienced neurologic, vascular, or visceral complications. There were 88 screws (30 thoracic and 58 lumbar) available for CT evaluation. The overall accuracy was 93.2% with 1 medial breach and 5 lateral breaches. The median screw diameter was 4.5 mm (range, 3.5-5.5 mm) for the thoracic spine and 5.5 mm (range, 4-6 mm) for the lumbar spine. CONCLUSION.: The results suggest that pedicle screws can be safely and accurately placed in young children. The overall accuracy of acceptable pedicle screw placement was 93.2% in patients younger than 8 years of age. Screw diameters ranging from 3.5 to 5.5 mm (for the thoracic spine) and 4 to 6 mm (for the lumbar spine) can be safely used in this population. PMID- 20010399 TI - Cervical spine motion generated with manual versus jackson table turning methods in a cadaveric c1-c2 global instability model. AB - STUDY DESIGN.: Cadaveric biomechanical study. OBJECTIVE.: To quantify spinal motion created by transfer methods from supine to prone position in a cadaveric C1-C2 global instability model. SUMMARY OF BACKGROUND DATA.: Patients who have sustained a spinal cord injury remain at high risk for further secondary injury until their spine is adequately stabilized. To date, no study has evaluated the effect of patient transfer methods from supine to prone position in the operating room, on atlantoaxial cervical spine motion. METHODS.: A global instability was surgically created at the C1-C2 level in 4 fresh cadavers. Two transfer protocols were tested on each cadaver. The log-roll technique entailed performing a standard 180 degrees log-roll rotation of the supine patient from a stretcher to the prone position onto the operating room Jackson table (OSI, Union City, CA). The "Jackson technique" involved sliding the supine patient to the Jackson table, securing them to the table, and then rotating them into a prone position. An electromagnetic tracking device registered motion between the C1 and C2 vertebral segments. Three different head holding devices (Mayfield, Prone view, and blue foam pillow) were also compared for their ability to restrict C1-C2 motion. Six motion parameters were tracked. Repeated measures statistical analysis was performed to evaluate angular and translational motion. RESULTS.: For 6 of 6 measures of angulation and translation, manual log-roll prone positioning generated significantly more C1-C2 motion than the Jackson table turning technique. Out of 6 motion parameters, 5 were statistically significant (P < 0.001-0.005). There was minimal difference in C1-C2 motion generated when comparing all 3 head holding devices. CONCLUSION.: The data demonstrate that manual log-roll technique generated significantly more C1-C2 motion compared to the Jackson table technique. Choice of headrest has a minimal effect on the amount of motion generated during patient transfer, except that the Mayfield device demonstrates a slight trend toward increased C1-C2 motion. PMID- 20010400 TI - Intraoperative computed tomography with integrated navigation system in spinal stabilizations. AB - STUDY DESIGN.: A prospective interventional case-series study plus a retrospective analysis of historical patients for comparison of data. OBJECTIVE.: To evaluate workflow, feasibility, and clinical outcome of navigated stabilization procedures with data acquisition by intraoperative computed tomography. SUMMARY OF BACKGROUND DATA.: Routine fluoroscopy to assess pedicle screw placement is not consistently reliable. Our hypothesis was that image guided spinal navigation using an intraoperative CT-scanner can improve the safety and precision of spinal stabilization surgery. METHODS.: CT data of 94 patients (thoracolumbar [n = 66], C1/2 [n = 12], cervicothoracic instability [n = 16]) were acquired after positioning the patient in the final surgical position. A sliding gantry 40-slice CT was used for image acquisition. Data were imported to a frameless infrared-based neuronavigation workstation. Intraoperative CT was obtained to assess the accuracy of instrumentation and, if necessary, the extent of decompression. All patients were clinically evaluated by Odom-criteria after surgery and after 3 months. RESULTS.: Computed accuracy of the navigation system reached <2 mm (0.95 +/- 0.3 mm) in all cases. Additional time necessary for the preoperative image acquisition including data transfer was 14 +/- 5 minutes. The duration of interrupting the surgical process for iCT until resumption of surgery was 9 +/- 2.5 minutes. Control-iCT revealed incorrect screw position >/=2 mm without persistent neurologic or vascular damage in 20/414 screws (4.8%) leading to immediate correction of 10 screws (2.4%). Control-iCT changed the course of surgery in 8 cases (8.5% of all patients). The overall revision rate was 8.5% (4 wound revisions, 2 CSF fistulas, and 2 epidural hematomas). There was no reoperation due to implant malposition. According to Odom-criteria all patients experienced a clinical improvement. A retrospective analysis of 182 patients with navigated thoracolumbar transpedicular stabilizations in the preiCT era revealed an overall revision rate of 10.4% with 4.4% of patients requiring screw revision. CONCLUSION.: Intraoperative CT in combination with neuronavigation provides high accuracy of screw placement and thus safety for patients undergoing spinal stabilization. Reoperations due to implant malpositions could be completely avoided. The system can be installed into a pre-existing operating environment without need for special surgical instruments. The procedure is rapid and easy to perform without restricted access to the patient and-by replacing pre- and postoperative imaging-is not associated with an additional exposure to radiation. Multidisciplinary use increases utilization of the system and thus improves cost efficiency relation. PMID- 20010401 TI - Reparative perineural hyperplasia and dermatofibroma. PMID- 20010402 TI - Squamoid eccrine ductal carcinoma: a case report and review of the literature. AB - Eccrine ductal carcinoma is a rare adnexal tumor. Squamoid eccrine ductal carcinoma (SEDC) represents an exceptionally rare variant with only 6 cases reported to date. This neoplasm is deeply infiltrative and may provide a diagnostic challenge in superficial shave biopsies of skin. We present a case of SEDC in an immunocompromised patient, who, after an initial shave biopsy for diagnostic purposes, underwent a complete excision of his tumor by Mohs micrographic surgery. A combined histologic and immunohistochemical approach is necessary to identify and confirm the unique characteristics of SEDC. Because this cancer is very rare and little is known about its biologic behavior, optimal diagnostic and treatment guidelines need to be developed. PMID- 20010403 TI - Melanotic neuroectodermal tumor of infancy presenting in the subcutaneous soft tissue of the thigh. AB - Melanotic neuroectodermal tumor of infancy (MNTI) is a rare and diagnostically challenging neoplasm typically presenting in the bones of the maxilla, skull, or mandible. Only 6 of approximately 357 reported cases have involved the subcutis. We describe a case of MNTI presenting as a palpable, subcutaneous, thigh mass in a 5-month-old girl. By ultrasound, the mass was round with well-defined borders, minimal vascularity, and heterogeneous echogenicity. Microscopically, the tumor consisted of nested foci of primitive-appearing small round blue cells with an increased nuclear to cytoplasmic ratio, stippled chromatin, and occasional mitotic figures. A larger and more epithelioid second cell population exhibited eosinophilic cytoplasm and sparse pigmented granules. The background stroma was fibrous and densely sclerotic. The differential diagnosis of soft tissue MNTI can include melanoma, neuroblastoma, rhabdomyosarcoma, desmoplastic small round cell tumor, and other pediatric "small round cell" neoplasms. The tumor had the characteristic immunophenotype of MNTI: cytokeratin+, HMB-45+, neuron-specific enolase+, and synaptophysin+. MNTI should be considered in the differential diagnosis of pigmented soft tissue lesions in children. Our patient remains disease-free 40 months after excision, although these tumors can locally recur (10%-20%) and rarely metastasize. PMID- 20010404 TI - Nodular mucinosis of the breast: expanding our understanding with an unusual case. AB - Nodular mucinosis of the breast is an extraordinarily rare lesion that occurs in patients who do not have Carney syndrome. Typically, the affected patients are young women with no medical history and a nodule under 1 nipple. Histopathologic examination has, until now, shown a multinodular myxoid lesion containing scattered capillaries and histiocytes but void of epithelial components. We present the case of a 72-year-old woman with a history of mucinous carcinoma of the breast who now presents with a painful subareolar nodule of the same breast. Biopsy and histological examination confirmed nodular mucinosis of the breast. PMID- 20010405 TI - When dead cells tell tales-cutaneous involvement by precursor T-cell acute lymphoblastic lymphoma with an uncommon phenotype. AB - The thymic type of precursor T-cell acute lymphoblastic lymphoma (pre-T ALL), an uncommon T-cell malignancy, typically presents as a thymic mass and expresses terminal deoxonucleotidyl transferase, CD7, and cytoplasmic CD3, with variable expression of other markers. Cutaneous presentation in pre-T ALL is highly unusual. We describe a case of pre-T ALL presenting as 2 papulonodular lesions on the face of an otherwise asymptomatic 27-year-old man. Microscopic examination of both lesions revealed a moderate to dense pandermal infiltrate of medium-sized lymphocytes with extensive "crush" artifact, whereas immunohistochemistry revealed positive staining of lesional cells for CD45, CD3, Bcl-2, Ki-67, CD5, CD7, and CD34 but negative staining for CD4, CD8, CD30, CD56, CD10, CD117, anaplastic lymphoma kinase protein, TdT, myeloperoxidase, CD79a, and CD20. Gene rearrangement studies performed on both biopsies identified a clonal population of T lymphocytes. A subsequent computed tomography scan revealed a 9-cm mediastinal mass encasing all major mediastinal vessels, whereas a bone marrow biopsy revealed blasts with an immunophenotype similar to that of the cutaneous lesions. Features unique to this case include the cutaneous presentation and the immunophenotype-absence of CD4, CD8, and TdT with expression of CD34-both highly unusual features for pre-T ALL. PMID- 20010406 TI - Cutaneous metastases of malignant melanoma: a clinicopathologic study of 192 cases with emphasis on the morphologic spectrum. AB - Metastatic melanoma represents one of the most common types of cutaneous metastases. In up to 5% of patients, metastatic melanoma can be the first manifestation of the disease. For the most part, the histologic diagnosis of metastatic melanoma poses little diagnostic difficulty; however, some metastases may adopt unusual or unfamiliar appearances mimicking other benign and malignant conditions. We present a study of 192 cases of cutaneous metastatic melanomas with special emphasis on their spectrum of morphologic features. The patients were 115 men and 77 women, 23-91 years of age (mean 57 years). Most tumors were located on the proximal legs, scalp, and arms and ranged from 0.8 to 3.0 cm. One hundred ten cases showed the classic morphologic appearance of melanoma (well circumscribed epithelioid dermal/subcutaneous nodule), 82 cases showed unusual histologic appearances that mimicked other benign and malignant neoplasms. In 16 patients (8.3%), there was no evidence of primary melanoma and the cutaneous metastasis was the only manifestation of the disease. The histologic diagnosis of cutaneous metastatic melanoma can pose difficulties for diagnosis, especially in the face of an unknown primary neoplasm. Unusual features observed in this series included examples of cutaneous metastatic melanoma that closely simulated metastatic carcinoma, dermatofibroma, leiomyosarcoma, angiosarcoma, nevoid melanoma, halo nevus, blue nevi, and atypical fibroxanthoma. Several cases also showed rhabdoid, balloon cell, and alveolar features. Immunohistochemical stains plus careful clinical history helped to establish the correct diagnosis. Our series illustrates that the differential diagnosis of cutaneous metastatic melanoma can be broad and difficult. To the best of our knowledge, this is the largest series of cutaneous metastatic melanomas reported in the literature. PMID- 20010407 TI - Naringenin inhibits neointimal hyperplasia following arterial reconstruction with interpositional vein graft. AB - Vessels respond to injury by a healing process that includes the development of neointima. Stenosis secondary to neointima formation is the main cause of failure following arterial reconstructions. Vessel wall homeostasis is regulated by proinflammatory cytokines that affect smooth muscle cell proliferation, growth, migration, and death. We assessed the hypothesis that naringenin, a flavinoid possessing anti-inflammatory, antioxidant, and antiproliferative activities, reduces neointimal hyperplasia (NIH) following vascular injury.Arterial injury was created by interposition grafting of autologous right superficial epigastric vein graft into the right femoral artery (FA) in 48 male Sprague-Dawley rats. Following injury, the rats were divided into 4 groups (n = 12). Two groups were treated with naringenin (100 mg/kg intraperitoneal q daily) for 2 and 4 weeks each while 2 control groups received normal saline for the same durations. For Sham group (n = 10), the FA and vein were isolated without any additional procedure. Rats were killed at the end of treatment regimen in all groups, and FAs were harvested. Thickness of intima was measured in histologic sections, and levels of platelet derived growth factor (PDGF)-BB, TNFalpha, and Ki67 labeling index (Ki67 LI) were quantified in immunohistochemical analyses to assess the amount of NIH and mechanisms underlying its formation.Although there was no significant difference between the groups at 2 weeks, neointima thickness was lower in the naringenin treated group at 4 weeks (23.7 +/- 2.3 vs. 35.6 +/- 2.6 microm in control group; P < 0.001). The levels of PDGF-BB, and TNFalpha were lower in naringenin treated groups at both 2 weeks (PDGF-BB [0.21% +/- 0.03% versus 0.39% +/- 0.05% in control group, P < 0.001), TNFalpha (21.2% +/- 0.8% vs. 36.1% +/- 1.9% in control group, P < 0.001]) and 4 weeks (PDGF-BB [0.25% +/- 0.03% vs. 0.57% +/- 0.09% in control group, P < 0.001], TNFalpha [25.5% +/- 1.8% vs. 45.0% +/- 2.9% in control group, P < 0.001]). Ki67 LI was lower in naringenin treated groups at 2 weeks (13.9% +/- 2.8% vs. 18.7% +/- 3.7% in control group, P < 0.05), and at 4 weeks (17.5% +/- 2.6% vs. 31.1% +/- 4.7% in control group, P < 0.001), indicating a lower level of cellular proliferation.Naringenin reduces NIH following arterial reconstruction. This may be mediated by a decrease in PDGF-BB and TNFalpha levels and the resulting down-regulation of smooth muscle cells' migration and proliferation. PMID- 20010408 TI - Superior gluteal artery perforator flap in bilateral breast reconstruction. AB - The incidence of bilateral breast reconstruction is increasing particularly due to genetic counseling and the option for bilateral prophylactic mastectomies. The decision to undergo a prophylactic mastectomy depends on the achievable outcomes of breast reconstruction. The free superior gluteal artery perforator flap (sGAP) flap is one option for autologous bilateral reconstruction which has rarely been reported.All bilateral sGAP flaps performed in the department of plastic surgery at the Behandlungszentrum Vogtareuth over a period of 4.5 years were retrospectively analyzed for indication, success rate, and complications.Thirty sGAP flaps were performed for bilateral breast reconstruction. The average age of the 15 women was 42 years and the average body mass index was 20.8. Indications for breast reconstruction were predominantly prophylactic mastectomies (60%). Indication for a sGAP flap was either a thin patient with insufficient abdominal tissue or a 2-staged bilateral reconstruction. 83% of the breast reconstructions were performed secondarily and 93% in 2 stages. The average operating time was 7 hours 12 minutes. Twenty-nine flaps (97%) were successful. Complications were fat necrosis (n = 3), hematoma (n = 3), and breast seroma (n = 1). Donor site complications were seroma (n = 8), infection (n = 1), and wound dehiscence (n = 1).Our results with bilateral breast reconstruction with the sGAP flap show cosmetically appealing results with high success and low complication rates on the breast. However, seromas on the donor site occurred in 27%. In addition, the sGAP flap is a technically demanding and time consuming operation. We recommend the sGAP flap when the abdomen has not enough tissue bulk to perform a deep inferior epigastric perforator flap or for a 2-staged bilateral reconstruction. This is often the case in women with a hereditary high risk of breast cancer who often present as young and slim patients. PMID- 20010409 TI - Management of chronic leg and knee pain following surgery or trauma related to saphenous nerve and knee neuromata. AB - Patients who present with lower extremity pain following surgery or trauma can occasionally have the saphenous nerve as the offending anatomic cause of their pain. Consistent with its anatomic course, the saphenous nerve can be the source of pain that manifests anywhere along its sensory distribution.Patients who presented to the Georgetown Peripheral Nerve Institute with lower extremity pain were evaluated, and those patients whose pain was suspected to be of saphenous nerve origin were offered surgical treatment. The surgical intervention included excision of the neuroma and/or nerve decompression, as clinically indicated. Patients were followed clinically and evaluated for both their pain as well as recovery in their ambulation and quality of life.Forty-two consecutive patients underwent surgery for pain of saphenous nerve origin from 2003 to 2008; 69% of these patients had concomitant surgery on another involved lower extremity peripheral nerve. Follow-up was achieved in 35 patients (83% response rate), with an average follow-up duration of 34.7 months. Using a 10-point pain scale, patients reported their preoperative pain as an 8.0 and their postoperative pain as a 2.37 (P < 0.001). Of the 35 patients, 30 (86%) were able to ambulate at the last follow-up encounter. Patients were asked to report their quality of life on a 10-point scale, and reported a 77% recovery of their baseline quality of life as a result of peripheral nerve surgery performed. Of the 35 patients, 29 reported that the surgery effectively resolved their pain, yielding a success rate of 82.8%.The saphenous nerve can be a source of lower extremity and knee pain following trauma or surgery. Accurate clinical diagnosis followed by surgical intervention can result in clinical resolution in the majority of patients, with improvement in ambulation and quality of life. This study reports the largest series of surgically-corrected saphenous neuropathy in the literature. PMID- 20010410 TI - The influence of surgical excision margins on keloid prognosis. AB - Keloid disease is known to have variable clinical behavior in response to therapy and there is no clinicopathologic classification that predicts such varied behavior. The aim of this study was to study the effect of excision margins and other histopathologic characteristics on keloid prognosis.Seventy-five multiethnic patients presenting with keloid scars at a department of plastic and reconstructive surgery during an 11-year period were included in this study. Clinical data was collected and detailed histologic analyses using light microscopy were carried out on archived patient specimens.A detailed histopathologic examination of all tissue samples identified keloid border or margin characteristics which were classified into "circumscribed" (borders clearly-demarcated) and "infiltrative" (borders not clearly-demarcated and not easily-definable). The specific histologic findings were correlated with keloid recurrence which revealed that incomplete peripheral (P < 0.001) and deep excision margins (P < 0.001), as well as infiltrative borders (P < 0.05) were associated with higher 1-year reported recurrence rates.This study has given evidence that incomplete surgical excision are associated with higher recurrence and this may justify the practice of routine histopathologic reporting of keloid excision margins. PMID- 20010411 TI - Hyalomatrix PA in burn care practice: results from a national retrospective survey, 2005 to 2006. AB - Results of a national retrospective survey on Hyalomatrix PA in burn patients are reported.A total of 11 burn centers were contacted.A total of 57 patients were available. Hyalomatrix PA was used on young and adult patients, mainly in deep partial thickness and full thickness burns. In most cases, Hyalomatrix PA was applied immediately after the wound cleaning (wound debridement or escharectomy for adults, dermabrasion or debridement in young patients). After 7 days, reepithelization processes were more frequent in deep partial thickness burns. One-half of the patient population underwent grafting. After 29 days, complete closure was achieved in almost all patients. The Vancouver Scar Scale showed better values for adults, while no differences were observed for burn depth or patients undergoing grafting. No adverse reactions were recorded.Hyalomatrix PA is used in young and adults, in deep partial thickness and full thickness burns, as a temporary coverage before grafting or alone for wound healing. PMID- 20010412 TI - Perineal reconstruction with bilateral bilobed flap for cloacal-like defect after childbirth. AB - Fourth degree lacerations of the vaginal wall and perineum following childbirth can cause cloacal-like defects of the perineum. The loss of the anterior anal sphincter and resultant thinning of the perineum and rectovaginal septum can lead to fecal incontinence and difficulties with sexual activity. This article introduces a technique to combine repair of the anterior anal sphincter with reconstruction of the perineal body with bilateral pedicled bilobed flaps. PMID- 20010413 TI - Asymmetric facial growth and deviated nose: a new concept. AB - Deviated nose correction is difficult and constitutes a very different issue from septal deviation. When correcting this deformity, traces of asymmetry can be detected. The authors demonstrate facial asymmetry accompanying deviated noses, and such asymmetry is usually ignored by surgeons who typically concentrate only on nose deformities.A total of 5822 pre- and postrhinoplasty photographs related to 547 women and 124 men were reviewed. Out of the total population, the following 3 groups were selected: group A, gross nose and face asymmetry; group B, nose asymmetry with no facial deformity; group C, facial asymmetry with straight nose. Different measurements were applied to the selected photos, presented in . These included measurement from the lateral canthi to the lateral mouth corners (D1) and from the midface to each most lateral part of the zygomatic arch (D2). Measurements from one side were compared with those from the contralateral side to identify true anatomic differences, as presented in .(Table is included in full-text article.)(Figure is included in full-text article.)There was a significant difference in the nose and face deformity group, as evidenced by a meaningful difference in both the D1 and D2 measurements.We report a significant growth retardation of the midface and orbit on the concave side of the nose. This impediment may serve as the etiology for many asymmetries of the face and nose. PMID- 20010414 TI - Prefabrication of tissue engineered bone grafts: an experimental study. AB - The purpose of this study was to determine whether angiogenesis could successfully be induced into bone tissue that was engineered by cultured adipose derived stem cells with porous beta-tricalcium phosphate and whether its biologic properties could be maintained by flap prefabrication technique.Adipose-derived stem cells with porous beta-tricalcium phosphate were implanted into the superficial inferior epigastric artery flap of the Fisher rats. After prefabrication for 8 weeks, the prefabricated flaps were elevated and the pedicles were clamped for 4 hours. The samples were harvested after 2 weeks for analyses.Angiogenesis was significantly increased in the prefabricated groups (P < 0.05). There was no significant difference between the prefabricated and nonprefabricated groups in terms of the osteogenic capacity (P > 0.5).The promising results obtained with prefabrication in tissue engineered bone grafts encourage the clinical application of this technology. Thus, prefabrication may be a useful technique in any engineered bone tissue transfer. PMID- 20010415 TI - Visual and pathologic analyses of keloid growth patterns. AB - Keloids grow and spread not only vertically but also horizontally, although hypertrophic scars do not grow beyond the boundaries of the original injury. Clinically, we have encountered keloids with regular and irregular (untypical) shapes. As the characteristics of the irregular growth patterns of keloids have not been studied yet, we analyzed the irregular growth of keloids both visually and pathologically.A total of 220 keloid specimens, each from a different patient, were surgically removed and used in this study. Through visual analysis, the preoperative shapes of these 220 keloids were classified into those with a regular shape (R group) and those with an irregular shape (IR group). Moreover, we distinguished between cases that had received keloidectomy previously and those that had not. We also determined whether the keloids were recurrent keloids or not. In both the R and IR groups, keloid specimens were studied histologically to examine for infection.In the R group, there were 156 cases (70.9%; 55 males and 101 females with a mean age of 33.68 years). Three patients (1.9%) had infection and 2 patients (1.3%) had undergone keloidectomy previously. In the IR group, there were 64 cases (29.1%; 24 males and 40 females with a mean age of 45.27 years). Thirty patients (46.9%) had infection and 24 patients (37.5%) had undergone keloidectomy previously. Statistically, the rates of infection and keloidectomy were significantly different between the R group and the IR group.Severe infection or operative history may be the cause of irregularly shaped keloids. Thus, in the absence of significant infection or a surgical history, the shape of keloids may be determined uniquely by skin tension. PMID- 20010416 TI - The vascular anatomy of the rat superficial epigastric flap by vascular corrosion casting and technical refinement for the study of choke vessels in cadaveric flap models. AB - Accurate depiction of cutaneous vascular microanatomy is of relevance to plastic surgical flap research, and to descriptive anatomy. Yet current techniques have not permitted full visualization of the subdermal plexus, or potential angiosomal connections. Nor has endothelial visualization been facilitated. Vascular corrosion casting techniques are promising in that regard, and were applied in an extended lateral thoracoabdominal suprafascial adipocutaneous flap in the rat (based on the superficial epigastric bundle). Technical refinements for application to further study of human cadaveric flap models are presented. The intraflap vascular branching pattern of the superficial epigastric artery is described, with filling of the lateral thoracic, intercostals, and iliolumbar angiosomes found when coagulation of vessels at the periphery was delayed until after clearance. The vascular casting protocol presented is an effective and promising tool for the study of macro- and microvascular anatomy. PMID- 20010418 TI - Improving postoperative pain management: what are the unresolved issues? PMID- 20010419 TI - Predicting success on the certification examinations of the American Board of Anesthesiology. AB - BACKGROUND: Currently, residency programs lack objective predictors for passing the sequenced American Board of Anesthesiology (ABA) certification examinations on the first attempt. Our hypothesis was that performance on the ABA/American Society of Anesthesiologists In-Training Examination (ITE) and other variables can predict combined success on the ABA Part 1 and Part 2 examinations. METHOD: The authors studied 2,458 subjects who took the ITE immediately after completing the first year of clinical anesthesia training and took the ABA Part 1 examination for primary certification immediately after completing residency training 2 yr later. ITE scores and other variables were used to predict which residents would complete the certification process (passing the ABA Part 1 and Part 2 examinations) in the shortest possible time after graduation. RESULTS: ITE scores alone accounted for most of the explained variation in the desired outcome of certification in the shortest possible time. In addition, almost half of the observed variation and most of the explained variance in ABA Part 1 scores was accounted for by ITE scores. A combined model using ITE scores, residency program accreditation cycle length, country of medical school, and gender best predicted which residents would complete the certification examinations in the shortest possible time. CONCLUSIONS: The principal implication of this study is that higher ABA/ American Society of Anesthesiologists ITE scores taken at the end of the first clinical anesthesia year serve as a significant and moderately strong predictor of high performance on the ABA Part 1 (written) examination, and a significant predictor of success in completing both the Part 1 and Part 2 examinations within the calendar year after the year of graduation from residency. Future studies may identify other predictors, and it would be helpful to identify factors that predict clinical performance as well. PMID- 20010420 TI - Acquisition of critical intraoperative event management skills in novice anesthesiology residents by using high-fidelity simulation-based training. AB - BACKGROUND: Early acquisition of critical competencies by novice anesthesiology residents is essential for patient safety, but traditional training methods may be insufficient. The purpose of this study was to determine the effectiveness of high-fidelity simulation training of novice residents in the initial management of critical intraoperative events. METHODS: Twenty-one novice residents participated in this 6-week study. Three hypoxemia and three hypotension scenarios were developed and corresponding checklists were validated. Residents were tested in all scenarios at baseline (0 weeks) and divided into two groups, using a randomized crossover study design. Group 1 received simulation-based training in hypoxemic events, whereas Group 2 was trained in hypotensive events. After intermediate (3 weeks) testing in all scenarios, the groups switched to receive training in the other critical event. Final testing occurred at 6 weeks. Raters blinded to subject identity, group assignment, and test date scored videotaped performances by using checklists. The primary outcome measure was composite scores for hypoxemia and hypotension scenarios, which were compared within and between groups. RESULTS: Baseline performance between groups was similar. At the intermediate evaluation, the mean hypoxemia score was higher in Group 1 compared with Group 2 (65.5% vs. 52.4%, 95% CI of difference 6.3-19.9, P < 0.003). Conversely, Group 2 had a higher mean hypotension score (67.4% vs. 45.5%, 95% CI of difference 14.6-29.2, P < 0.003). At Week 6, the scores between groups did not differ. CONCLUSIONS: Event-specific, simulation-based training resulted in superior performance in scenarios compared with traditional training and simulation-based training in an alternate event. PMID- 20010421 TI - Incidence, Reversal, and Prevention of Opioid-induced Respiratory Depression. AB - Opioid treatment of pain is generally safe with 0.5% or less events from respiratory depression. However, fatalities are regularly reported. The only treatment currently available to reverse opioid respiratory depression is by naloxone infusion. The efficacy of naloxone depends on its own pharmacological characteristics and on those (including receptor kinetics) of the opioid that needs reversal. Short elimination of naloxone and biophase equilibration half lives and rapid receptor kinetics complicates reversal of high-affinity opioids. An opioid with high receptor affinity will require greater naloxone concentrations and/or a continuous infusion before reversal sets in compared with an opioid with lower receptor affinity. The clinical approach to severe opioid induced respiratory depression is to titrate naloxone to effect and continue treatment by continuous infusion until chances for renarcotization have diminished. New approaches to prevent opioid respiratory depression without affecting analgesia have led to the experimental application of serotinine agonists, ampakines, and the antibiotic minocycline. PMID- 20010422 TI - Camera phone as the medium. PMID- 20010423 TI - Exertional rhabdomyolysis and malignant hyperthermia in a patient with ryanodine receptor type 1 gene, L-type calcium channel alpha-1 subunit gene, and calsequestrin-1 gene polymorphisms. PMID- 20010424 TI - Myelotoxicity of oral topotecan in relation to treatment duration and dosage: a phase I study. AB - Oral topotecan has been recently brought into clinical practice at a dose of 2.3 mg/m(2) for 5 days, every 3 weeks. Published data show quite high myelotoxicity. The aim of this trial was to define the daily dose and treatment duration, which permits safe toxicity. The study was designed to begin at a low daily dosage of 1.5 mg/m(2) and was escalated by increasing the topotecan dose and the day treatment duration. The plan was to end up with 2.3 mg/m(2) daily for 5 days. In cases of tolerability with the last dosage given, we would then continue testing a higher daily dosage. Treatment repetition was planned to be every 21 days. Dosage levels were 1.5, 2.0 and 2.3 mg/m(2) for 3 days, 2.0 and 2.3 mg/m(2) for 4 days, and 2.3 mg/m(2) for 5 days. Toxicity was scored according to the Common Toxicity Criteria. Thirty-two patients (27 male, five female, median age 60 years, range 46-77 years) with small-cell lung cancer were included. The patients on 1.5 and 2 mg/m(2) for 3 days showed no myelotoxicity. Four (25%) patients on 2.3 mg/m(2) 3-day treatment developed grade 3-4 neutropenia. Three of five patients (60%) treated for 4 days at a dose of 2.3 mg/m(2) developed grade 3-4 neutropenia and less than half (two of five, 40%) of these patients had thrombocytopenia. Eight patients (66.7%) on the 5-day treatment presented with serious grade 3-4 myelotoxicity. Two treatment-related deaths were observed in the 5-day group and one in the 4-day group. Granulocyte growth factor was applied in over 60% of the patients. In conclusion, a dose of 2.3 mg/m(2) for 5 days was intolerable. Dose-limiting toxicity was 2.3 mg/m(2) for 4 days without prophylactic granulocyte colony-stimulating factor administration. The safe duration of oral topotecan treatment and the maximum tolerated dose seem to be not longer than 3 days at a dose of 2.3 mg/m(2). PMID- 20010425 TI - The potential of resveratrol against human gliomas. AB - There is growing interest in dietary phytochemicals as potential cancer chemopreventive agents. Resveratrol (3,4',5-trihydroxy-trans-stilbene), a naturally occurring phytoalexin that is present in grapes, red wine, berries and peanuts, has been studied extensively for its ability to interfere with multistage carcinogenesis. Resveratrol is known to have antioxidant, anti inflammatory and antiproliferative effects on a variety of cancer cells in vitro and in various animal models. However, the effect(s) of resveratrol in vivo on humans are still controversial. This study discusses current knowledge with regard to the effects of resveratrol in relation to its potential as a chemopreventive and/or chemotherapeutic molecule against human gliomas. PMID- 20010426 TI - Targeting the efficacy of a dendrimer-based nanotherapeutic in heterogeneous xenograft tumors in vivo. AB - Our earlier studies have shown the in vitro and in vivo targeting of a generation 5 (G5) dendrimer-based multifunctional conjugate that contained folic acid (FA) as the targeting agent and methotrexate (MTX) as the chemotherapeutic drug. To clinically apply the synthesized G5-FA-MTX nanotherapeutic, it is important that the anticancer conjugate elicits cytotoxicity specifically and consistently. Toward this objective, we evaluated the large-scale synthesis of a G5-FA-MTX conjugate (Lot # 123-34) for its cytotoxic potential and specificity in vitro and in vivo. The cytotoxicity and specificity were tested by using a coculture assay in which FA receptor-expressing and nonexpressing cells (KB and SK-BR-3 cells, respectively) were cultured together and preferential killing was examined. The in-vitro data were compared with the in-vivo data obtained from a heterogeneous xenograft tumor model. The animal model of the artificial heterogeneous xenograft tumor showed that the nanotherapeutic was preferentially cytotoxic to KB cells. PMID- 20010427 TI - Predictors of progression from prehypertension to hypertension among rural Chinese adults: results from Liaoning Province. AB - BACKGROUND: Individuals with prehypertension are at a high risk of developing hypertension. Little is known about the predictors of progression from prehypertesion to hypertension in rural Chinese adults. METHODS: A population based sample of 15 061 rural Chinese aged > or =35 years with prehypertension free cardiovascular diseases at baseline were followed-up from 2004-2006 to 2008. Incident hypertension was defined as systolic blood pressure (BP) > or =140 mmHg, diastolic BP > or =90 mmHg, or current use of antihypertensive medications. RESULTS: During the 28-month follow-up period, 4917 individuals (32.6%) with prehypertension developed hypertension, in which only 1.5% (n = 73) of incident cases reached BP control target. After Cox stepwise regression analysis, baseline age (per 5 years) [hazard ratio (HR): 1.111; 95% confidence interval (CI): 1.095 1.126], Mongolian (HR: 1.079; 95% CI: 1.010-1.152), alcohol drinking, overweight and obese (HR: 1.349; 95% CI: 1.261-1.444), salt intake everyday, inappropriate physical activity, and family history of hypertension were associated with incident hypertension independent of initial systolic and diastolic BP. Among these predictors, current alcohol drinking (odds ratio: 1.126; 95% CI: 1.049 1.209), salt intake, and overweight and obese were associated only with increased systolic BP, whereas inappropriate physical activity contributed only to increase diastolic BP. CONCLUSION: These data suggest a high incidence of hypertension in individuals with prehypertension. Lifestyle modifications such as cessation of drinking, healthy diet, and moderate physical activity are advised to prevent or delay the progression of prehypertension to hypertension in rural areas of China. PMID- 20010428 TI - Y25130 hydrochloride, a selective 5HT3 receptor antagonist has potent antimitogenic and apoptotic effect on HT29 colorectal cancer cell line. AB - Serotonin (5-hydroxytryptamine, 5-HT) is known to be a mitogenic factor in several malignancies. It elicits its mitogenic effect through a wide range of 5 HT receptor subtypes and several internal cellular transcription pathways. According to wide distribution of 5-HT3 and 5-HT4 receptors in the gastrointestinal tract, the main aim of this study was to investigate the effect of these receptor agonists and antagonists in a colorectal cell line. In cell culture, we investigated the effects of 5-HT, 5-HT3 and 5-HT4 receptor agonists and antagonists on proliferation of HT29 cells. We also tested apoptosis for the receptor antagonists with TUNEL apoptosis test. In addition, we assayed effects of 5-HT receptor antagonists on cell cycle kinetics with flow cytometery. Proliferation assay revealed that phenylbiguanide (a 5-HT3 receptor selective agonist) increased proliferation of HT29 cells significantly and Y25130 hydrochloride (a 5-HT3 receptor antagonist) had the opposite effect; but for 5 HT4 receptor antagonist, these effects were not significant. In addition, potent apoptotic and cell cycle arresting effect was found for a selective 5-HT3 receptor antagonist but we have not seen any significant effect for the 5-HT4 receptor antagonist. The findings of this study provide strong evidence for the potential role of 5-HT3 receptors in colorectal cancer. PMID- 20010430 TI - The potential efficacy of 3,3'-diindolylmethane in prevention of prostate cancer development. AB - The objective of this study was to examine the efficacy of 3,3'-diindolylmethane (DIM) in prevention of prostate cancer tumor development in an animal model. Mouse prostate cancer cells (TRAMP-C2, 2x10) were injected subcutaneously into three groups of C57BL/6 mice (10 mice in each group). Two groups were treated earlier with DIM; 2 or 10 mg/kg each, and an additional control group was injected with medium. Animals were treated for five more weeks until sacrificed. Tumor sizes were measured biweekly. At the end of the experiment, mice were sacrificed, and tumors were excised, weighed, measured and tested using immunohistochemical studies. In addition blood samples were collected for biochemical analysis. The results indicated that DIM significantly reduced tumor development in treated animals when compared with controls. Tumors developed in 80% of controls and 40% and 60% of animals treated with 10 or 2 mg/kg of DIM, respectively. Moreover, tumors that developed in treated animals were significantly (P<0.001) smaller than in controls. Additionally, our results indicated that DIM has no effect on animal weight or liver and kidney functions. These results indicated that the DIM agent is not toxic and has an in-vivo preventive effect against the development of prostate cancer in a mouse model. PMID- 20010429 TI - A pooled analysis of interval cancer rates in six European countries. AB - The objective of this study was to assess detection rates and interval breast cancer (IC) rates from eight programmes in the European Breast Cancer Screening Network. A common data collection protocol was used to explore differences in IC rates among programmes and discuss their potential determinants. Pooled analysis was used to describe IC rates by age, compliance in screening, recall rate, screening detection (SD) rate and expected breast cancer incidence. Participation in screening averaged 77.9% (range 42.6-88.7%), recall rate 5.4% (range 3.3 17.7%) in the initial and 3.4% (range 1.8-8.9%) in the subsequent screening rounds, and SD rate was 60.4 (range 41.6-91) per 10 000 women in initial and 38.5 (range 31.3-62.6) in subsequent screens. IC rate during first 12 months after screening was 5.9 (range 2.1-7.3) per 10 000 women screened negative and 12.6 (range 6.3-15) in the second year of the interval. IC comprised 28% of the IC and SD cancers. The ratio between IC rate and expected incidence was 0.29 for the first 12 months and 0.63 for the 13-24 months period. Sensitivity was higher for the ages 60-69 years and for initial tests than subsequent tests. There were distinct differences in the IC rates between programmes. The results of this study reveal large variations in screening sensitivity and performance. Pooled evaluation of some process indicators within the European breast cancer screening programmes proved to be feasible and is likely to be useful for the future, particularly if it is performed regularly and extensively. PMID- 20010431 TI - Dispatch codes of out-of-hospital cardiac arrest should be diagnosis related rather than symptom related. AB - OBJECTIVE: To describe the characteristics and outcome in out-of-hospital cardiac arrest (OHCA) in relation to (i) whether OHCA was coded by the dispatcher as a diagnosis or as a symptom and (ii) the delay until the first unit was alerted at the dispatch centre. METHODS: OHCA patients in Goteborg, during 17 months, excluding OHCA after calling the rescue team. RESULTS: Among 250 cases, 20% were coded as a diagnosis (i.e. CA) with or without ongoing cardiopulmonary resuscitation (CPR). Dispatch codes for the remaining 200 patients (80%) were mostly symptom related (unconsciousness in 61%, codes related to breathing problems in 10%, other codes in 24% and missing in 5%). Patients in whom the dispatchers coded the call as CA had an earlier start to CPR after collapse (median 2 vs. 10 min; P<0.0001) and a higher rate of bystander CPR (86% vs. 42%; P<0.0001). Furthermore, they tended to have a higher rate of survival to hospital discharge (14.0% vs. 6.5%; P = 0.09). The median delay until the first unit was alerted was 1.8 min. Survival to hospital discharge was 10.0% if the delay was below median and 6.7% if the delay was above median (P = 0.48). CONCLUSION: Patients with OHCA who were not coded by dispatchers as such had a long delay to the start of CPR and a low survival. Dispatching according to diagnosis, that is, CA seems to improve these parameters most likely reflecting a more optimal communication between the dispatcher and the caller as well as the rescue team. PMID- 20010432 TI - Lower limbs function and pain relationships after unilateral total knee arthroplasty. AB - The aim of the study was to evaluate gait characteristics, lower limbs joint function, and pain relationships associated with knee osteoarthritis of female patients before and 3 months after total knee arthroplasty at an outpatient clinic rehabilitation department. Gait parameters were registered, the active range of lower extremity joints was measured, and pain was evaluated. Gait characteristics, walking velocity, and knee joint's extension improved and pain diminished considerably 3 months after arthroplasty, nevertheless did not reach control group level. In conclusion, single support phase and gait velocity change reflect the recovery of gait function after total knee arthroplasty. Beside the affected knee joint's range of motion, attention should be paid to the restoration of the functional reserve of the adjacent joints and joints of contralateral lower extremity after total knee arthroplasty. Affected knee joint pain perceived during level walking influenced the gait velocity of patients before total knee arthroplasty but not 3 months after arthroplasty. PMID- 20010434 TI - Pravastatin counteracts angiotensin II-induced upregulation and activation of NADPH oxidase at plasma membrane of human endothelial cells. AB - Endothelial dysfunction has been linked to reactive oxygen species (ROS) production by nicotinamide adenine dinucleotide phosphate reduced (NADPH) oxidase. Angiotensin II (ANG), which levels are elevated in some cardiovascular diseases, can stimulate this enzyme, whereas statins have been demonstrated pleiotropic effects related with the restoration of endothelial function. Therefore, our purpose was to study the mechanism of the possible beneficial effects of pravastatin on ANG-activated human umbilical vein endothelial cells (HUVEC). ANG induced an increase in the extracellular superoxide anion produced by NADPH oxidase but had no effect in the intracellular ROS production. Pravastatin, which alone did not have any effect on ROS production, totally blocked the stimulating effects of ANG when combined with it. These effects were not due to a direct action of ANG or pravastatin on the activity of NADPH oxidase measured in HUVEC lisates. On the contrary, the results correlated well with other effects of ANG: a Nox4 and p22phox upregulated expression and an enhanced Nox4 translocation to the cell membrane. All these effects were inhibited by pravastatin, which had no effect when incubated alone. These data reveal for the first time that pravastatin interrupts the signaling pathway activated by ANG that leads to an enhanced NADPH oxidase activity at the cell membrane of HUVEC. For that, pravastatin inhibits ANG-induced upregulation of catalytic NADPH oxidase subunits and blocks the migration of them to the endothelial cell membrane. PMID- 20010433 TI - HIV-1 viral subtype differences in the rate of CD4+ T-cell decline among HIV seroincident antiretroviral naive persons in Rakai district, Uganda. AB - BACKGROUND: Data on the effect of HIV-1 viral subtype on CD4 T-cell decline are limited. METHODS: We assessed the rate of CD4 T-cell decline per year among 312 HIV seroincident persons infected with different HIV-1 subtypes. Rates of CD4 decline by HIV-1 subtype were determined by linear mixed effects models, using an unstructured convariance structure. RESULTS: A total of 59.6% had D, 15.7% A, 18.9% recombinant viruses (R), and 5.8% multiple subtypes (M). For all subtypes combined, the overall rate of CD4 T-cell decline was -34.5 [95% confidence interval (CI), -47.1, -22.0] cells/ microL per yr, adjusted for age, sex, baseline CD4 counts, and viral load. Compared with subtype A, the adjusted rate of CD4 cell loss was -73.7/microL/yr (95% CI, -113.5, -33.8, P < 0.001) for subtype D, -43.2/microL/yr (95% CI, -90.2, 3.8, P = 0.072) for recombinants, and 63.9/microL/yr (95% CI, -132.3, 4.4, P = 0.067) for infection with multiple HIV subtypes. Square-root transformation of CD4 cell counts did not change the results. CONCLUSIONS: Infection with subtype D is associated with significantly faster rates of CD4 T-cell loss than subtype A. This may explain the more rapid disease progression for subtype D compared with subtype A. PMID- 20010436 TI - Homocysteine thiolactone induces cardiac dysfunction: role of oxidative stress. AB - This study investigates the cardiac functioning in male Wistar rats after treatments with methionine and homocysteine thiolactone (HcyT). The rats were distributed into 3 groups and treated for 8 weeks. Group I was the control (CO) group, given water, group II was treated with methionine, and group III with HcyT (100 mg/kg). Morphometric and functional cardiac parameters were evaluated by echocardiography. Superoxide dismutase (SOD), catalase, and glutathione S transferase activities, chemiluminescence, thiobarbituric acid reactive substances, and immunocontent were measured in the myocardium. Hyperhomocysteinemiawas observed in rats submitted to the both treatments. The results showed diastolic function was compromised in HcyT group, seen by the increase of E/A (peak velocity of early (E) and late (A) diastolic filling) ratio, decrease in deceleration time of E wave and left ventricular isovolumic relaxation time. Myocardial performance index was increased in HcyT group and was found associated with increased SOD immunocontent. HcyT group demonstrated an increase in SOD, catalase, and glutatione S-transferase activity, and chemiluminescence and thiobarbituric acid reactive substances. Overall, these results indicated that HcyT induces a cardiac dysfunction and could be associated with oxidative stress increase in the myocardium. PMID- 20010435 TI - Preconditioning by subinotropic doses of ouabain in the Langendorff perfused rabbit heart. AB - Short exposure to low concentrations of digitalis drugs like ouabain protects the rat heart against ischemia/reperfusion injury through the activation of the Na/K adenosine triphosphatase (ATPase)/Src receptor complex and subsequent stimulation of key intracellular cardioprotective signals. Rat Na/K-ATPase, however, is relatively insensitive to digitalis, and it is not known if similar results could be obtained in species with higher sensitivity. Thus, to determine whether ouabain pretreatment protects against ischemic injury and activates the Na/K ATPase signaling cascade in a species with cardiac glycoside sensitivity comparable to humans, the present study was conducted in the rabbit model. In Langendorff perfused rabbit hearts, 20-minute exposure to 500-nM ouabain resulted in positive inotropy as evidenced by a significant increase in +dP/dt, and this increase was accompanied by the activation of several well-characterized downstream mediators of the cardiac Na/K-ATPase receptor pathway, including Src, Akt, ERK1/2, and protein kinase Cepsilon. A short (4 minutes) administration of a subinotropic dose of ouabain (100 nM) followed by an 8-minute washout before 30 minutes of global ischemia and 120 minutes of reperfusion resulted in protection against cell death, as evidenced by a significant decrease in infarct size. These data indicate that ouabain administration activates the Na/K-ATPase signaling cascade and protects against ischemic injury in a species with high cardiac Na/K ATPase sensitivity. PMID- 20010438 TI - The role of uncoupling protein 2 in the apoptosis induced by free fatty acid in rat cardiomyocytes. AB - The apoptotic loss of cardiomyocytes contributes to numerous cardiovascular disorders. Evidence suggests that free fatty acids induce cellular apoptosis, and recent studies have shown that free fatty acids dramatically elevate mRNA levels of uncoupling protein 2 (UCP2) in some cell lines. In this study, we investigated the possibility that free fatty acids induce the expression of UCP2 through the peroxisome proliferator-activated receptor pathway, thereby increasing cell apoptosis in adult rat cardiomyocytes. Primary cultured adult rat cardiomyocytes exposed to free fatty acids exhibited a dose-dependent increase in apoptosis. Quantitative real-time reverse transcription-polymerase chain reaction and Western blotting showed significant increases in the level of UCP2 expression at 6, 12, and at 24 hours after treatment of adult rat cardiomyocytes with free fatty acids. Expression of UCP2 was suppressed with RNA interference, and knockdown of UCP2 attenuated free fatty acid-induced apoptosis in the cardiomyocytes. In summary, free fatty acids induced UCP2 expression through peroxisome proliferator-activated receptor alpha in adult rat cardiomyocytes. PMID- 20010437 TI - Na/H exchange inhibition protects newborn heart from ischemia/reperfusion injury by limiting Na+-dependent Ca2+ overload. AB - The results of the Guardian/Expedition trials demonstrate the need for more precisely controlled studies to inhibit Na/H exchange (NHE1) during ischemia/reperfusion. This is because overwhelming evidence is consistent with the hypothesis that myocardial ischemic injury results in part from increases in intracellular Na (Nai) mediated by NHE1 that in turn promote Na/Ca exchanger mediated increases in intracellular Ca ([Ca]i) and Ca-dependent cell damage. We used a more potent and specific NHE1 inhibitor HOE 694 (HOE) to test whether inhibition of NHE1 during ischemia limits increases in Nai and [Ca]i in newborns. NMR was used to measure pHi, Nai, [Ca]i, and ATP in isolated newborn rabbit hearts. Perfusion pressure, left ventricular developed pressure, and creatine kinase were measured. HOE was added before global ischemia. Results are reported as mean +/- SE. Nai (mEq/kg dry weight) rose from 11.6 +/- 0.9 before ischemia to 114.0 +/- 16.1 at the end of ischemia and recovered to 55.2 +/- 11.8 in the control group. During ischemia and reperfusion, the corresponding values for Nai in the HOE group (63.1 +/- 8.4 and 15.9 +/- 2.5, respectively, P < 0.05) were lower than control. In the control group [Ca]i (nM/L) rose from 331 +/- 41 to 1069 +/- 71 and recovered to 814 +/- 51, whereas in the HOE group [Ca]i rose less (P < 0.05): 359 +/- 50, 607 +/- 85, and 413 +/- 40, respectively. Total creatine kinase release was significantly reduced in the HOE group. Perfusion pressure and left ventricular developed pressure also recovered significantly better in the HOE group than in the control. In conclusion, NHE1 inhibition diminishes ischemia induced increases in Nai and therefore [Ca], and thus diminishes myocardial injury in neonatal hearts. PMID- 20010439 TI - Cytochalasin D, LY294002 and olomoucine synergize in promoting death of melanoma cells through activation of caspase-3 and apoptosis. AB - Many of the current anticancer therapies rely on the induction of apoptosis, and several mechanisms that protect cells against apoptosis may be upregulated in tumors. A growing body of evidence suggests that single drugs with a clearly defined intracellular target may be less efficient in arresting tumor growth and induction of apoptosis than multitargeted strategies. To prove that this is also the case for melanoma, we studied five cell lines, which represent different stages of tumor progression. We tested cell viability, terminal dUTP nick-end labeling and activation of caspase-3 upon exposure to cytochalasin D, LY294002 and olomoucine, added either alone or in various combinations. The obtained data were compared with effects caused by staurosporine. The results show that whereas staurosporine efficiently induced apoptosis in all tested melanoma cell lines, the other drugs had only moderate effects when administered alone. In contrast, the combinations of drugs were more effective in inducing caspase-3 activity and reducing cell viability. In particular, the triple combination of cytochalasin D+LY294002+olomoucine was almost as effective as staurosporine in inducing caspase-3 activity and apoptosis. These results prove that it is possible to design new pharmacological strategies that will effectively induce caspase-3 activity and apoptosis in melanoma. The possible explanations of the observed synergy between the tested drugs are also discussed. PMID- 20010440 TI - Primary malignant melanoma of the esophagus. AB - Primary malignant melanoma of the esophagus is an extremely rare, but highly aggressive, tumor. The prognosis for primary malignant melanoma of the esophagus remains dismal in most literatures. We report here a case of primary malignant melanoma of the esophagus that was treated by surgical resection and, additionally, followed by chemotherapy. The clinical features, treatment, pathological findings, and prognosis are analyzed and the literature of primary malignant melanoma of the esophagus is reviewed. PMID- 20010441 TI - Can we predict who will experience recurrent vasomotor symptoms after stopping hormone therapy? PMID- 20010442 TI - Fronto-parietal overactivation in patients with essential tremor during Stroop task. AB - Neuropsychological dysfunctions have been consistently reported in essential tremor but the underlying neurobiological mechanisms are unknown. We explored potential abnormalities in the neural network involved in cognitive functions in patients with essential tremor by using functional magnetic resonance imaging. The functional response of 12 patients with essential tremor and 12 matched controls was studied while performing a functional magnetic resonance imaging Stroop task aimed to assess attentional control and evaluating executive functions. Despite similar performances during this task, patients with essential tremor showed greater magnitude of brain response in the dorsolateral prefrontal cortex and in the inferior parietal cortex with respect to controls. Our study shows that patients with essential tremor require additional cognitive effort to achieve comparable performance levels on test of attentional control. PMID- 20010443 TI - An event-related potential study of temporal information encoding and decision making. AB - This research used event-related potentials to investigate the time course of temporal information encoding and decision making. Participants performed auditory temporal and pitch discrimination tasks within a S1-S2 matching paradigm, wherein two stimuli (S1-S2) are presented sequentially, and S2 is either the same as or different from S1. Event-related potentials results showed N1, P2 and contingent negative variation (CNV) amplitudes were higher during encoding than decision making. During the encoding phase, N1 and P2 amplitudes were lower in the temporal task than the pitch task. During the decision-making phase, the CNV amplitude was higher in the temporal task. Findings suggest N1, P2 and CNV are related to temporal encoding, albeit only CNV is related to temporal decision making. PMID- 20010444 TI - State sadness reduces neural sensitivity to nonrewards versus rewards. AB - Both behavioral and neural evidence suggests that depression is associated with reduced sensitivity to rewards. Using the feedback negativity, a neural index of reward processing, an earlier study showed that depressive symptoms experienced over the previous week were associated with less differentiation between nonrewards and rewards in a gambling task. To directly test whether variability in state mood related to similar effects on neural correlates of reward, this study recorded the feedback negativity in individuals assigned to either a neutral or sad mood induction. Following the induction, individuals reporting greater sadness exhibited a reduced feedback negativity. This finding indicates that fluctuation in state negative affect moderates how environmental feedback is processed by reducing neural sensitivity to nonrewards versus rewards. PMID- 20010445 TI - Evidence of a gustatory-vestibular pathway for protein transport. AB - OBJECTIVE: To demonstrate anatomically a pathway for protein transport from the palate to the vestibular system. METHOD: The vestibulofacial anastomosis and associated ganglion cells were identified in a collection of 160 horizontally sectioned human temporal bones that had been stained with hematoxylin and eosin. Wheat germ agglutinin-horseradish peroxidase (HRP) was applied to the greater superficial petrosal nerve in 4 Sprague-Dawley rats. After 30 hours, the rats were killed by intracardiac perfusion, and the seventh and eighth nerves with adjacent brainstem removed. Frozen sections cut at 30 mum through this block were then reacted for HRP, counterstained with neutral red, and mounted on slides for examination in the light microscope. RESULTS: Thirty-two of the 160 human temporal bones contained sections through the vestibulofacial anastomosis and its ganglion. In all cases, the ganglion was incorporated into the vestibular ganglion (VG) adjacent to the nervus intermedius. In all 4 experimental rats, HRP reaction product labeled a small number of ganglion cells in the VG adjacent to the nervus intermedius and facial nerve. CONCLUSION: These observations support the presence of a pathway from receptors in the palate to the VG. PMID- 20010446 TI - Topical dexamethasone and tympanic membrane perforation healing in otitis media: a short-term study. AB - BACKGROUND: The safety and efficacy of topical corticosteroids have not been established in the treatment of acute otitis media (AOM) with tympanic membrane (TM) perforation. Specific concerns include the risk for nonhealing TM perforations. Previous studies have not been done in models with concurrent AOM. The purpose of this study was to determine whether the addition of dexamethasone to topical ciprofloxacin affects healing of AOM-related TM perforations. METHODS: Bilateral AOM was induced in chinchillas by transbulla injection of Streptococcus pneumoniae, and TMs were perforated with a CO2 laser 3 to 5 days later. Each animal received ciprofloxacin-dexamethasone in one ear and ciprofloxacin in the other, twice daily for 7 days. Tympanic membranes were assessed for 4 weeks. RESULTS: Fifteen of 55 ciprofloxacin-treated and 37 of 55 ciprofloxacin dexamethasone-treated TMs failed to heal within the 4-week study period (p < 0.001). Exposure to dexamethasone increased the odds for persistent perforation by 5.5 (95% confidence interval, 2.4-12.6). At 4 weeks, mean TM perforation diameter was 0.9 mm in ciprofloxacin-treated ears and 2.1 mm in ciprofloxacin dexamethasone-treated ears (p < 0.001). CONCLUSION: Topical dexamethasone hinders TM healing in a chinchilla model of AOM at short-term follow-up. Longer follow-up is needed. PMID- 20010448 TI - High-grade undifferentiated sarcoma of the cerebellopontine angle masquerading as a benign vestibular schwannoma. PMID- 20010449 TI - Fluoxetine response in impulsive-aggressive behavior and serotonin transporter polymorphism in personality disorder. AB - BACKGROUND: Disturbances in central serotonin function have been implicated in impulsive and aggressive behavior. A deletion/insertion polymorphism within the 5 HT transporter promoter gene (5-HTTLPR) is thought to be associated with disturbed impulse control, anxiety, and depression. The serotonin transporter (5 HTT) is the primary action site for selective serotonin reuptake inhibitors (SSRIs). Several studies of major depression have shown that the l allele of 5 HTTLPR is associated with better SSRI antidepressant effects than the s allele. METHODS: This study investigates the association between response of impulsivity to treatment with fluoxetine and 5-HTTLPR polymorphism in 49 personality disordered patients. Additionally, we studied TPH1, 5HT1B and 5HT2C receptor polymorphisms as predictors of response in this population. RESULTS: Results reveal that patients with the l/l genotype of 5-HTTLPR had a significantly better response to fluoxetine when compared to s allele carriers, as evaluated on the basis of total (P<0.05) and Aggression subscale (P<0.01) Overt Aggression Scale Modified-score percentage change. There were no significant associations between fluoxetine response and TPH1 (A218C) (-6525 A>G) (-5806 G>T), HTR1B (G861C) and HTR2C (G68C) genotype groups. CONCLUSION: This is the first study assessing the association between these polymorphisms and anti-impulsive response to fluoxetine in personality disorder. As the s genotype is associated with a poorer selective serotonin reuptake inhibitors response in major depression, bulimia nervosa and borderline personality disorder, it could represent a common biological background for SSRI response. PMID- 20010450 TI - A genetic variant of HTR2C may play a role in the manifestation of Tourette syndrome. AB - Gilles de la Tourette syndrome (GTS) (MIM 137580) is a complex neuropsychiatric disorder probably originating from a disturbed interplay of several neurotransmitter systems in the prefrontal-limbic-basal ganglia loop. Polygenetic multifactorial inheritance has been postulated; nevertheless, no confirmed susceptible genes have been identified yet. As neuroimaging studies allude to dopaminergic and serotonergic dysfunction in GTS and serotonin as an important factor for dopamine release, genotyping of common polymorphisms in the serotonergic receptor (HTR1A: C-1019G; HTR2A: T102C, His452Tyr, A-1438G; HTR2C: C 759T, G-697C) and transporter genes (SLC6A4) was carried out in 87 patients with GTS, compared with 311 matched controls. We found a nominally significant association between both polymorphisms in the HTR2C and the GTS, which was more pronounced in male patients. Analysis of the further serotonergic polymorphisms did not reveal any significant result. A modified function of these promoter polymorphisms may contribute to the complex interplay of serotonin and dopamine and then to the manifestation of GTS. PMID- 20010451 TI - Angiotensin-converting enzyme polymorphism in schizophrenia, bipolar disorders, and their first-degree relatives. AB - BACKGROUND: Family, twin and adoption studies have provided major evidence for the role of genetics in numerous psychiatric disorders including schizophrenia (SZ) and bipolar disorders (BDs). As SZ and BD have some susceptibility genes in common and since unaffected first-degree relatives of these patients carry a high likelihood of these susceptibility genes, we aimed to elucidate the role of angiotensin-converting enzyme (ACE) genetic variants in patients with SZ, BD and their first-degree relatives. METHODS: The study sample comprised 239 patients with SZ, 184 patients with BD, 284 unaffected first-degree biological relatives of patients with SZ and 301 unaffected first-degree biological relatives of patients with BD and 210 healthy controls. The ACE genotypes were determined by polymerase chain reaction. RESULTS: ACE insertion/deletion polymorphism was associated with SZ and BD. DD genotype and D allele distributions in bipolar patients and their first-degree relatives were significantly higher than those of SZ patients, their relatives, and controls. In contrast, II genotype and I allele were reduced in both the patient groups and their relatives as compared with controls. CONCLUSION: In this study, the D allele might be responsible for clustering of psychotic symptoms and results in the psychotic manifestations of BD, whereas I allele seems to be protective against development of SZ and BD. SZ and BD characterized by similar or different gene variant in ACE could be a useful marker for these psychiatric disorders, if this polymorphism is replicated in the future studies. PMID- 20010452 TI - Retinal vessel analyzer measurements of the effect of panretinal photocoagulation on the retinal arteriolar diameter in diabetic retinopathy. AB - PURPOSE: The purpose of this study was to investigate the effect of panretinal photocoagulation (PRP) on the retinal arteriolar diameter in patients with diabetic retinopathy using a retinal vessel analyzer. METHODS: Ten eyes of 6 consecutive patients with type II diabetes and severe nonproliferative or proliferative diabetic retinopathy were studied prospectively. Measurements of the retinal arteriolar diameter were performed before the first photocoagulation session and after the end of the PRP treatment. RESULTS: Retinal arteriolar diameter before PRP was 131 +/- 15 arbitrary units and decreased to 112 +/- 14 arbitrary units after PRP (P = 0.012). There was a significant vasoconstriction of 13.8% +/- 8.3% following PRP. Mean visual acuity before and after PRP was 0.31 +/- 0.36 logarithm of the minimal angle of resolution and 0.28 +/- 0.30 logarithm of the minimal angle of resolution, respectively (P = 0.68). There was no significant change in mean arterial pressure before and after PRP (P = 0.89). There was no correlation between the visual acuity change or the number of laser burns and the percentage change in the retinal arteriolar diameter (P > 0.1). CONCLUSION: Panretinal photocoagulation has a vasoconstrictive effect on retinal arterioles in patients with severe nonproliferative or proliferative diabetic retinopathy. These results are consistent with an autoregulatory response of the retinal circulation to increased inner retinal oxygen tension after PRP. The retinal vessel analyzer is a fast, accurate, noninvasive, online measuring system for the study of the retinal vascular response to PRP in patients with diabetic retinopathy. PMID- 20010453 TI - Microperimetric retinal changes in myopic choroidal neovascularization treated with intravitreal ranibizumab. AB - PURPOSE: The purpose of this study was to report functional and morphologic retinal changes after intravitreal injections of 0.5 mg ranibizumab in patients with myopic neovascular membrane (choroidal neovascularization). METHODS: This is a case review of 11 consecutive patients with myopic choroidal neovascularization who received intravitreal injections of ranibizumab at monthly intervals. Serial changes in best-corrected visual acuity, optical coherence tomography, fluorescein angiography, and microperimetry (Nidek MP1, Nidek, Padova, Italy) are presented. RESULTS: Mean baseline best-corrected visual acuity was 59.82 + or - 17.50 Early Treatment Diabetic Retinopathy Study letters. After a follow-up of 36 weeks, mean visual acuity improved to 66.72 + or - 17.3 with a mean change of 6.91 letters. Mean baseline retinal sensitivity was 6.02 + or - 1.9 dB. After 36 weeks, mean sensitivity improved to 8.3 + or - 2.4 dB with a mean change of 2.3 dB. The fixation stability improved from 45.5% to 72.7%. All patients also had complete resolution of subretinal fluid, mean optical coherence tomography central retinal thickness was reduced from 244.64 + or - 39.3 microm to 191.36 + or - 27.3 microm at 36 weeks, and fluorescein angiography at 36 weeks showed absence of leakage in all patients. CONCLUSION: Intravitreal ranibizumab injections seemed to positively influence retinal functional status in patients with myopic choroidal neovascularization. PMID- 20010454 TI - Central retinal vein occlusion in a young Chinese population: risk factors and associated morbidity and mortality. AB - PURPOSE: The purpose of this study was to assess the risk factors for central retinal vein occlusion and associated morbidity and mortality in a Chinese population. METHODS: The participants included patients with central retinal vein occlusion 40 years old and younger. Predisposing factors, mortality, and systemic complications were examined in this group. RESULTS: Unilateral (n = 19) and bilateral (n = 3) central retinal vein occlusions were identified in a total of 22 patients (25 eyes), with a mean follow-up time of 37 months. Hypercholesterolemia (65%), hypertriglyceridemia (64%), and hyperhomocysteinemia (42%) were all identified as risk factors. Three patients (14%) developed stroke and 1 (5%) developed transient ischemic attacks during follow-up. Renal failure and pulmonary hypertension resulted in the death of two patients. The mean initial and final visual acuities (+ or - standard deviation) were 20/400 (+ or - 20/250) and 20/500 (+ or - 20/320), respectively, and treatments did not result in vision improvement (P = 0.57). The poor visual prognosis was likely due to macular edema in 7 eyes (28%), optic atrophy in 4 eyes (16%), and secondary glaucoma in 3 eyes (12%). CONCLUSION: Morbidity and mortality are high in young Chinese patients with central retinal vein occlusion who may have associated serious complications including stroke, blindness, and death. Central retinal vein occlusion may serve as an initial clinical presentation of serious systemic diseases. PMID- 20010455 TI - Retinopathy of prematurity and maternal age. AB - PURPOSE: To determine the risk factors, especially maternal risk factors, associated with the development of retinopathy of prematurity (ROP) in premature babies. METHODS: A matched case-control study involving premature patients was undertaken retrospectively. The case group consisted of premature babies with the subsequent development of ROP. The control group consisted of gestational age matched and sex-matched premature babies that did not develop ROP during the follow-up period. Risk factors involving patient demographics and maternal characteristics were compared between the case and control groups. RESULTS: A total of 144 patients were included in this study (72 patients in the case group and 72 patients in the control group). Among the 66 possible risk factors compared, only birth weight and maternal age were found to be significant risk factors. Birth weight was significantly lower in the case group (1,248.7 +/- 257.8 g vs. 1,335.5 +/- 297.2 g, P = 0.01), and maternal age was significantly older in the case group compared with that in the control group (31.2 +/- 5.1 years vs. 28.2 +/- 5.3 years, P < 0.001). The odds ratio of having babies with ROP was 2.9 when the maternal age was >30 years. CONCLUSION: Older maternal age is a newly identified risk factor for the development of ROP in premature babies. PMID- 20010456 TI - Central serous chorioretinopathy and phosphodiesterase-5 inhibitors: a case control postmarketing surveillance study. AB - PURPOSE: The purpose of this study was to determine if there is an increased risk of central serous chorioretinopathy (CSC) associated with prescription exposure to phosphodiesterase-5 (PDE-5) inhibitors. METHODS: A case-control study linking 2 National Veterans Health Administration databases (clinical and pharmacy) for fiscal years 2004 to 2005. The likelihood of past exposure to PDE-5 inhibitors among newly diagnosed patients with CSC, identified through International Classification of Diseases, 9th Edition, Clinical Modification codes, was compared with 2 age-matched control groups after excluding subjects with risk factors for CSC. RESULTS: Among 577 men, aged 59 years and younger with newly diagnosed CSC during the study year, 111 were prescribed a PDE-5 inhibitor (19.2%). The proportions of age-matched controls prescribed a PDE-5 inhibitor in the 2 groups were 18.5% and 21.5%. The odds ratio of exposure was 1.05 (95% confidence limit: 0.74-1.22) and 0.87 (95% confidence limit: 0.68-1.12). CONCLUSION: Patients with CSC had no increase in prescription exposure to PDE-5 inhibitors than did age-matched control subjects. Although the findings in this study do not support an association between CSC and PDE-5 inhibitors, postmarketing surveillance methods for drug-related side effects have acknowledged limitations. PMID- 20010457 TI - High-resolution melting analysis of sequence variations in the cytidine deaminase gene (CDA) in patients with cancer treated with gemcitabine. AB - Gemcitabine (2',2'-difluorodeoxycytidine) is a major antimetabolite cytotoxic drug with a wide spectrum of activity against solid tumors. Hepatic elimination of gemcitabine depends on a catabolic pathway through a deamination step driven by the enzyme cytidine deaminase (CDA). Severe hematologic toxicity to gemcitabine was reported in patients harboring genetic polymorphisms in CDA gene. High-resolution melting (HRM) analysis of polymerase chain reaction amplicon emerges today as a powerful technique for both genotyping and gene scanning strategies. In this study, 46 DNA samples from gemcitabine-treated patients were subjected to HRM analysis on a LightCycler 480 platform. Residual serum CDA activity was assayed as a surrogate marker for the overall functionality of this enzyme. Genotyping of three well-described single nucleotide polymorphisms in coding region (c.79A>C, c.208G>A and c.435C>T) was successfully achieved by HRM analysis of small polymerase chain reaction fragments, whereas unknown single nucleotide polymorphisms were searched by a gene scanning strategy with longer amplicons (up to 622 bp). The gene scanning strategy allowed us to find a new intronic mutation c.246+37G>A in a female patient displaying marked CDA deficiency and who had an extreme toxic reaction with a fatal outcome to gemcitabine treatment. Our work demonstrates that HRM-based methods, owing to their simplicity, reliability, and speed, are useful tools for diagnosis of CDA deficiency and could be of interest for personalized medicine. PMID- 20010458 TI - Comparison of a new serum topiramate immunoassay to fluorescence polarization immunoassay. AB - Topiramate is a newer anticonvulsant used to treat epilepsy, migraines, bipolar disorder, posttraumatic stress, and other conditions. Serum topiramate concentrations are measured to determine optimal levels, address therapeutic failure or drug-drug interactions, and assess compliance. Two high-throughput assays for serum topiramate measurement were compared: the Seradyn fluorescence polarization immunoassay (FPIA) on an Abbott TDx/FLx instrument and a new immunoassay from ARK Diagnostics performed on an Olympus AU680 automated analyzer. Precision, linearity, limit of quantitation, carryover, spike recovery, and endogenous interferences were found to be acceptable for the ARK assay. These studies were complemented by comparison of 120 patient samples analyzed using both methods. The ARK immunoassay performed comparably to FPIA with minimal difference in serum topiramate concentrations within the therapeutic range (2.0 20 microg/mL). A slight systematic discordance was observed at higher concentrations (greater than 30 microg/mL) with ARK immunoassay results being on average 6% higher than FPIA. Thus, the ARK immunoassay appears to provide acceptable analytical performance and comparability to FPIA; furthermore, the assay is compatible with high-throughput autoanalyzers. PMID- 20010459 TI - CYP3A5*3 genotype associated with intrasubject pharmacokinetic variation toward tacrolimus in bioequivalence study. AB - Tacrolimus is metabolized by CYP3A and has highly variable pharmacokinetics. To study the factors contributing to this high variability in pharmacokinetics and to investigate the possibility of genotype-specific clinical applications, the effect of differing CYP3A5 genotypes on the intrasubject coefficients of variation for tacrolimus was investigated. Genotyping for CYP3A5*3 was performed in healthy volunteers who had previously participated in the pharmacokinetic study of 2 tacrolimus formulations with a 2 x 2 cross-over design. Intrasubject coefficients of variation calculated from analysis of variation in CYP3A5*1/*1+*1/*3 (n = 16) and CYP3A5*3/*3 (n = 13) groups were compared. The intrasubject CVs of AUClast and Cmax in the CYP3A5*3/*3 group were about 41.1% and 52.4% greater than those in the CYP3A5*1*1+*1/*3 group. The estimated total sample size for the bioequivalence study of tacrolimus with a 2 x 2 cross-over design was increased by 93.3% for AUClast (n = 30 versus 58) and 121.4% for Cmax (n = 28 versus 62) in the CYP3A5*3/*3 group compared with the CYP3A5*1/*1+*1/*3 group. The intraindividual variability of tacrolimus PK parameters may be associated with the CYP3A5 genotype. We propose that genotyping for CYP3A5 will provide a more efficient approach for bioequivalence designs and therapeutic drug monitoring. PMID- 20010460 TI - Ultra fast liquid chromatography-tandem mass spectrometry routine method for simultaneous determination of cyclosporin A, tacrolimus, sirolimus, and everolimus in whole blood using deuterated internal standards for cyclosporin A and everolimus. AB - Specific chromatographic methods for the measurement of cyclosporin A, tacrolimus, sirolimus, and everolimus blood levels in patients with organ transplants are time consuming when large numbers of samples must be processed. The authors developed a robust and fast (1 minute) online solid-phase extraction liquid chromatography/tandem mass spectrometry method for the simultaneous quantification of cyclosporin A, tacrolimus, sirolimus, and everolimus. After protein precipitation of the whole blood with zinc sulphate and methanol, the supernatant was loaded on a wide pore reversed-phase column and cleansed of potential interferences with high flow for 20 seconds. After column switching, the analytes were transferred within 20 seconds in the back-flush mode to a short phenyl-hexyl column. The valve was then returned to its initial position and the chromatographic separation performed within 20 seconds. In the meantime, the loading column was prepared for the next injection. Ammoniated adducts of protonated molecules were used as precursor ions for all analytes. Multiple reaction mode transitions for each immunosuppressant and the internal standards were used for quantification. The working range of the method was 10-1500 microg/L for cyclosporin A, 1.0-44 microg/L for tacrolimus, 1.0-48 microg/L for sirolimus, and 1.2-48 microg/L for everolimus. Within and between-run assay coefficients of variation ranged from 1.8% to 13.0%. The described liquid chromatography/tandem mass spectrometry method shows best performance using the internal standards cyclosporin A-d4 for cyclosporin A, everolimus-d4 for everolimus and ascomycin for tacrolimus and sirolimus. In conclusion, the authors present a very fast, robust, and economical analytical method for therapeutic monitoring of multiple immunosuppressants in daily clinical practice. PMID- 20010461 TI - The European society of cardiology 2009 guidelines on the prevention, diagnosis, and treatment of infective endocarditis: key messages for clinical practice. PMID- 20010462 TI - Serum human cartilage glycoprotein-39 levels in patients with systemic lupus erythematosus. AB - INTRODUCTION: Human cartilage glycoprotein-39 (HC gp-39) is a protein secreted by various cells including chondrocytes. Serum HC gp-39 has been suggested to be a marker of cartilage damage. However, inflammation involving other sites than the joints is an additional factor that increases the serum level of HC gp-39. OBJECTIVES: The aim of the study was to evaluate the usefulness of HC gp-39 determination in serum of patients with systemic lupus erythematosus (SLE) as a marker of joint involvement. PATIENTS AND METHODS: Serum HC gp-39 levels were measured in 25 patients with SLE and 22 healthy controls. SLE activity was assessed by the Systemic Lupus Erythematosus Disease Activity Index, and articular involvement by calculating the number of swollen and tender joints. The markers of inflammation (erythrocyte sedimentation rate, C-reactive protein) were determined. RESULTS: We observed an increase in HC gp-39 in SLE patients. However, there was no correlation of this parameter with disease activity, inflammatory markers (except serum gamma globulin levels), and articular involvement. CONCLUSIONS: The study suggests that increased HC gp-39 in SLE patients results mainly from inflammation and is not useful as a marker of joint involvement. PMID- 20010463 TI - Superoxide anion production by bronchoalveolar lavage cells in relation to cellular composition and lung function in sarcoidosis and chronic bronchitis. AB - INTRODUCTION: Increased generation of superoxide anion (O(2)(-).) by bronchoalveolar lavage (BAL) cells has been reported in various inflammatory disorders. However, the clinical relevance of this phenomenon is unclear. OBJECTIVES: The aim of the study was to investigate whether production of O(2)( ). is enhanced in smoking-related chronic bronchitis and sarcoidosis, and to assess a relationship between O(2)(-). generation and lung function impairment and changes in BAL cellular pattern. PATIENTS AND METHODS: Forty-two patients with sarcoidosis, 24 smokers with chronic bronchitis, and 17 controls were examined. A number/percentage of BAL cells was calculated. Spontaneous and phorbol myristate acetate (PMA)-stimulated O(2)(-). production was measured in BAL cells. Spirometry was performed. RESULTS: Patients with smoking-related chronic bronchitis produced more O(2)(-). spontaneously (6.42 -/+1.24 vs. 15.39 /+2.47 nmol/106 cells, P = 0.003) and after stimulation (3.73 -/+1.32 vs. 14.76 /+2.79 nmol/106 cells; P = 0.001). PMA-stimulated excess production correlated with the percentage of neutrophils (r = 0.66, P = 0.0005). In sarcoidosis, only spontaneous production of O(2)(-). was higher (vs. 18.07 -/+2.49 nmol/106 cells, P = 0.004) and correlated with the percentage of BAL lymphocytes. There was no correlation between O(2)(-). production and lung function parameters. CONCLUSIONS: Patients with smoking-related chronic bronchitis produce more O(2)( )., and this phenomenon is related to BAL neutrophils. In sarcoidosis, spontaneous release of O(2)(-). from BAL cells is related to the extent of lymphocytic alveolitis. Higher O(2)(-). generation did not impair lung function. PMID- 20010464 TI - Frequency of BCR-ABL gene mutations in Polish patients with chronic myeloid leukemia treated with imatinib: a final report of the MAPTEST study. AB - INTRODUCTION: The presence of BCR-ABL oncogene mutations in patients with chronic myeloid leukemia (CML) may be responsible for the failure of tyrosine kinase inhibitor treatment. OBJECTIVES: The aim of the study was to evaluate the frequency of BCR-ABL gene mutations in patients with CML (the MAPTEST study) treated with imatinib (IM). PATIENTS AND METHODS: Direct sequencing analysis of BCR-ABL gene was performed in 92 patients treated with IM for more than 3 months. The mean time of IM treatment was 18 months. At the time of the analysis, 75 patients were in the first chronic phase (CP), 4 in the second CP, 5 in the acceleration and 8 in the blastic phase. Fifty-seven patients (62%) were treated with IM at a daily dose of 400 mg and 35 patients with higher doses (600 or 800 mg daily). Inclusion criteria were based on the European Leukemia Net definitions for failure and suboptimal response to IM. RESULTS: Twelve mutations were detected in 11 of 92 patients, including 4 mutations (36.7%) diagnosed during CP, 3 (27.3%) in acceleration, and 4 (36.7%) in blast crisis. In 1 patient with lymphoid blast crisis of CML coexisting F359V and Y253F mutations were detected. In the whole group mutations were detected in 2 of 5 patients (40%) with primary resistance (M351T, F359V + Y253F) and in 9 of 87 patients (10.3%) (E255K, T315I 3x, M351T, E355G, F359V-2x) with acquired resistance to IM. CONCLUSIONS: The study confirmed the usefulness of BCR-ABL gene mutation screening in patients with CML resistant to IM therapy. PMID- 20010465 TI - Persistent pain in the older adult: what should we do now in light of the 2009 American geriatrics society clinical practice guideline? AB - The recent publication of revised guidelines for the management of persistent pain in the older adult (American Geriatric Society, 2009) has posed a dilemma for clinicians. In essence, these revised guidelines now downplay the use of nonsteroidal anti-inflammatory drugs (NSAIDs) relative to prior year's recommendations. The strong recommendation for caution when employing NSAIDs is because of the numerous, well-documented, potential adverse effects including renal failure, stroke, hypertension, heart failure exacerbations, and gastrointestinal complications. Nevertheless, physicians still have a substantial arsenal for combating chronic pain due to such conditions as degenerative arthritis and back problems. Options for intervention include physical therapy, topical nonsteroidals, capsaicin, topical lidocaine, intra-articular therapies, and judicious use of narcotics. In the future, cyclooxygenase-inhibiting nitric oxide-donating drugs may represent a technical improvement in the toxicity profile of traditional NSAIDs. PMID- 20010466 TI - Practical implications of the revised guidelines for inpatient glycemic control. AB - Substantial observational data has linked hyperglycemia in hospitalized patients with poor patient outcomes. While early studies suggested improved clinical outcomes with interventions targeting near euglycemia, more recent studies have yielded inconsistent results, with the suggestion of harm with more severe hypoglycemia. The American Association of Clinical Endocrinologists and American Diabetes Association published a revised consensus statement on inpatient glycemic management that takes into account this recent evidence. This statement identifies reasonable, achievable, and safe glycemic targets and describes protocols, procedures, and system improvements necessary to achieve these effectively. These modified glycemic targets promote a rational approach to inpatient glycemic management that minimizes risks associated with uncontrolled hyperglycemia and hypoglycemia. Intravenous insulin infusions are recommended for critically ill patients who experience blood glucose (BG) levels above 140 mg/dl with a target of 140 to 180 mg/dl. Lower BG targets (i.e., 110-140 mg/dl) may be appropriate for patients following cardiac or vascular surgical procedures. In noncritically ill patients, scheduled subcutaneous basal:bolus insulin is the preferred therapy for achieving fasting and preprandial BG below 140 mg/dl and random BG values below 180 mg/dl, with consideration of more or less stringent targets based on a patient's clinical status. Prolonged use of correctional insulin as monotherapy is discouraged. Oral and injectable noninsulin glucose lowering agents have a limited role for hospital use but may be appropriate for selected noncritically ill patients. Educating personnel about appropriate inpatient glycemic management practices, obtaining reliable and reproducible measures of BG, and careful implementation of standardized protocols can help to ensure patient safety. PMID- 20010467 TI - Peritoneal dialysis and its role in the demography and epidemiology of chronic kidney disease. AB - In a population of patients who have undergone renal replacement therapy, the 21st century brought a rise in the number of elderly people as well as patients with obesity, with renal complications of systemic diseases, especially diabetes and hypertension, and a constantly increasing number of patients with congestive heart failure who require intermittent or permanent dialysis support. In keeping with the concept of an integrated approach to renal replacement therapy, peritoneal dialysis should be the first option in renal replacement treatment, especially in patients with residual renal function. This paper summarizes the current evidence supporting the view that the changing demography and epidemiology of chronic kidney disease imposes an introduction of patient friendly modifications of therapeutic options available in these patient groups. PMID- 20010468 TI - Peritoneal dialysis as a therapeutic approach in congestive heart failure resistant to pharmacological treatment. AB - Given an increasing number of patients with congestive heart failure (CHF) refractory to diuretics, new and more effective therapeutic modalities are sought. Peritoneal dialysis (PD), which provides continuous, slow ultrafiltration, may be an alternative to hemodialysis in this population. The current paper, based on a comprehensive literature review, addresses the role of PD in improving the quality of life of patients with CHF. PMID- 20010469 TI - Residual iliofemoral thrombosis in a 31-year-old woman on oral contraceptives possessing heterozygous factor V Leiden, factor XIII Val34Leu allele and alpha fibrinogen Thr312Ala allele: case report. AB - We present the case of a 31-year-old woman on oral contraceptives with a 3-year history of iliofemoral thrombosis resistant to recanalization despite satisfactory anticoagulation therapy and absence of concomitant diseases. Thrombophilia screening revealed heterozygous factor V Leiden mutation. We also detected the presence of factor XIII (FXIII) Leu34 allele and alpha-chain fibrinogen 312Ala allele, which are known to adversely affect fibrin clot structure and lysis. It might be speculated that the presence of 3 polymorphisms in this patient could contribute to proximal thrombosis resistant to treatment. We postulate that determination of FXIII and alpha-fibrinogen polymorphisms can be useful in the evaluation of some young patients with deep vein thrombosis. PMID- 20010470 TI - Kikuchi-Fujimoto disease: case report. AB - We present the case of a 27-year-old woman with local cervical lymphadenopathy without other symptoms. Due to a gradual progression of lesions, a biopsy was performed, and a specimen excised from an enlarged lymph node was subjected to histological examination. Kikuchi-Fujimoto disease (KFD), that is histiocytic necrotizing lymhadenitis, was diagnosed. Lymphadenopathy resolved without medical treatment over 6 months. KFD should be considered in the differential diagnosis of lymphadenopathy, even though this disease rarely occurs in Poland. PMID- 20010471 TI - Atypical clinical presentation of Churg-Strauss syndrome with rapidly progressive glomerulonephritis: case report. AB - The case of a 48-year-old man presented in this paper illustrates an atypical clinical course of Churg-Strauss syndrome with rapidly progressive glomerulonephritis with no signs of bronchial asthma. PMID- 20010472 TI - Peritoneal dialysis as a therapeutic approach in congestive heart failure resistant to pharmacological treatment: case report. AB - This report describes the use of continuous peritoneal dialysis (PD) as an alternative to hemodialysis (HD) in a patient with type 2 cardiorenal syndrome in the course of congestive heart failure resistant to standard pharmacological treatment. A 39-year-old man presented with a 24-year history of progressive heart failure. Ineligibility for heart transplant and previous inefficient treatment with different modifications of HD reduced his treatment options to PD. After 7 months of continuous PD (1 overnight exchange with icodextrin and 2 daily standard continuous ambulatory PD exchanges) his overall condition significantly improved compared with his status while on HD. An increase from NYHA class IV to class II, increase in left ventricular ejection fraction from 50% to 55%, decrease in right ventricular systolic pressure from 73 to 53 mmHg, and improvement in the quality of life enabled him to resume his daily activities. PMID- 20010473 TI - A diagnostically difficult case of chronic myeloid neoplasm with eosinophilia and abnormalities of PDGFRA effectively treated with imatinib in accelerated phase: case report. AB - Chronic myeloid neoplasm with eosinophilia and abnormalities of platelet-derived growth factor receptor alpha (PDGFRA), referred to until 2008 as chronic eosinophilic leukemia, is distinguished from hypereosinophilic syndrome (HES), if accompanied by genetic abnormalities that enable to determine eosinophil clonality. Typically, HES has a benign course and glucocorticosteroids suffice to achieve remission. In chronic myeloid neoplasm with eosinophilia and abnormalities of PDGFRA the FIP1L1-PDGFRA fusion gene can be detected. Its product is a protein showing tyrosine kinase activity leading to malignant proliferation of eosinophil precursors. Differential diagnosis of HES is often difficult because hypereosinophilia may also be reactive and may occur in many nonhematological as well as hematological disorders. Thus, reverse-transcription polymerase chain reaction (RT-PCR)is indicated in all patients with HES in order to detect the FIP1L1-PDGFRA transcript. Traditional treatment of chronic myeloid neoplasm with cytostatic drugs results in a short-term and transient remission or stabilization of the disease. We present the case of a 52-year-old patient with chronic myeloid neoplasm with eosinophilia and abnormalities of PDGFRA, in whom acceleration occurred after a year of cytostatic therapy with hydroxyurea and was successfully treated with imatinib. It was impossible to unequivocally determine the type of bone marrow disease based on histologic criteria, and a wide spectrum of molecular tests differentiating the type of myeloid proliferation were necessary to establish the diagnosis. RT-PCR did not reveal BCR-ABL or JAK2 V617F mutation. Further molecular testing showed rearrangement involving the FIP1L1 gene, thus enabling implementation of targeted therapy. PMID- 20010479 TI - Clinical guidelines for the management of acute viral infections in patients with systemic lupus erythematosus. AB - In recent decades, many research groups have focused on the role of viral infections in the etiopathogenesis of systemic lupus erythematosus (SLE), the so called "viral hypothesis". The main candidates are herpes viruses such as Epstein Barr virus (EBV) and cytomegalovirus (CMV), which have a high seroprevalence in the general population. However, a viral causal agent of SLE has not yet been discovered, although many interesting clinical findings on the complex interactions between viruses and SLE have been made. This review analyzes 88 cases of acute viral infections in adult patients with SLE and identifies situations in which viral infections influenced the diagnosis, prognosis or treatment of SLE. We also propose clinical guidelines for the management of these infections in patients with SLE. PMID- 20010480 TI - Therapy of chronic hepatitis B: new goals and new treatments. AB - Treatment of chronic hepatitis B has shown a rapid development in the last years leading to a shift of treatment strategies from interferon to hepatitis B virus (HBV)-polymerase inhibitors. In particular, treatment with HBV-polymerase inhibitors has changed the indication on how to treat a patient and when to stop therapy. Long-term treatment with HBV-polymerase inhibitors may often be required, even if it raises the possibility of resistance and subsequent treatment failure. This review provides a strategy on how to manage HBV therapy with the currently available treatment options. PMID- 20010481 TI - Treatment of chronic hepatitis C. AB - Treatment for chronic hepatitis C has changed over the past years achieving higher response rates. The combination treatment with pegylated interferon-a and ribavirin is tailored based on the on-treatment virological responses. With this response-guided therapy, the overall sustained virological response rate is about 55%. Many new antivirals are currently under investigation and some will be commercially available in the near future. These include antiviral molecules acting directly against the hepatitis C virus (HCV) replication machinery, such as the inhibitors of the viral protease, and agents binding to host cofactors of the viral replication, thereby inhibiting HCV in an indirect way (such as cyclophilin inhibitors and nitazoxanide). The advent of these drugs will further ameliorate response rates and facilitate the permanent cure of chronic hepatitis C. PMID- 20010482 TI - Viral infections in exacerbations of asthma and chronic obstructive pulmonary disease. AB - Respiratory viral infections are recognized as the most frequent cause of asthma and chronic obstructive pulmonary disease (COPD) exacerbations with rhinovirus (i.e. the virus of the common cold) being the most frequent identified virus. The recent development of human experimental models of rhinovirus-induced asthma and COPD exacerbations represent innovative tools with the potential to increase our understanding in this field. Moreover this models will provide the opportunity to test, in a carefully controlled setting, novel pharmacological compounds. In this review we will provide an overview of the role of viral infections in asthma and COPD exacerbations and in particular we will summarize the inflammatory and immunological mechanisms that can pave the way to exacerbation following respiratory viral infection in these patients. PMID- 20010483 TI - Viral infections in transplant recipients. AB - Solid organ and hematopoietic stem cell transplant recipients are uniquely predisposed to develop clinical illness, often with increased severity, due to a variety of common and opportunistic viruses. Patients may acquire viral infections from the donor (donor-derived infections), from reactivation of endogenous latent virus, or from the community. Herpes viruses, most notably cytomegalovirus and Epstein Barr virus, are the most common among opportunistic viral pathogens that cause infection after solid organ and hematopoietic stem cell transplantation. The polyoma BK virus causes opportunistic clinical syndromes predominantly in kidney and allogeneic hematopoietic stem cell transplant recipients. The agents of viral hepatitis B and C present unique challenges particularly among liver transplant recipients. Respiratory viral illnesses due to influenza, respiratory syncytial virus, and parainfluenza virus may affect all types of transplant recipients, although severe clinical disease is observed more commonly among lung and allogeneic hematopoietic stem cell transplant recipients. Less common viral infections affecting transplant recipients include those caused by adenoviruses, parvovirus B19, and West Nile virus. Treatment for viruses with proven effective antiviral drug therapies should be complemented by reduction in the degree of immunosuppression. For others with no proven antiviral drugs for therapy, reduction in the degree of immunosuppression remains as the sole effective strategy for management. Prevention of viral infections is therefore of utmost importance, and this may be accomplished through vaccination, antiviral strategies, and aggressive infection control measures. PMID- 20010484 TI - Prophylactic HPV vaccination: a major breakthrough in the fight against cervical cancer? AB - Cervical cancer is the second most common female cancer with 500000 new cases and 290000 deaths occurring worldwide per annum. Organized cervical screening programs have reduced the incidence and mortality of cervical cancer. However, in developing countries scarce resources, poverty, lack of infrastructure and disenfranchisement of women have been major hurdles in the effective implementation of routine screening programmes. As a result, 83% of cervical cancers still occur in the developing countries and account for 15% of all female cancers. Epidemiological studies have established a causative role of Human Papillomavi-rus (HPV) infection in the development of cervical cancer. The development and implementation of a prophylactic HPV vaccine will have a major impact on preventing this global disease. However, long-term surveillance of the HPV vaccination program will be required to confirm the expected reduction in cervical cancer incidence. This article reviews the role of HPV in the development of cancer and the burden of HPV related cancers; types and pharmacokinetics of HPV vaccines; challenges and issues in implementing vaccination programmes; screening in the developing and developed countries and screening options in the post-vaccination era. PMID- 20010485 TI - [GA(2)LEN (Global Allergy and Asthma European Network): European network of excellence for asthma and allergic diseases]. AB - Allergic diseases represent some of the main health problems in Europe. These are increasing in prevalence, seriousness and social cost. The Global Allergy and Asthma European Network (GA(2)LEN), a network of excellence of the 6 degrees management program, was created in the 2005 with the aim to gather the European leader institutions of the research and clinical assistance fields, in order to guarantee the excellence and avoid the fragmentation of the energy spent in fighting allergy diseases in general. The GA(2)LEN has drawn a great advantage from the personal efforts of every single researcher who have proved their strong motivation in carrying on this "pan-European" model of collaboration. The network has been organized in order to increase the team work in scientific research projects in allergic and asthma disease field, making the GA(2)LEN the worldwide leader in this area. On these basis research projects have been carried on about which first data have been already published. The activities of the GA(2)LEN include in general the establishment of a lasting organization of the planning phase, the activity linked to every single project and to the improving on the existing projects, as well as the draft of new guidelines. This review reports the main achieved goals. PMID- 20010486 TI - The study of polymorphism within the promoter region of the osteopontin (OPN) gene in sows. AB - OBJECTIVES: The aim of the present study was to find a possible polymorphism in the promoter region of the osteopontin (OPN) gene, as a potential mutation region, connected with the transcription factor-binding sites or regulatory sequences and to estimate the expression of this gene in ovaries and oviduct of sows. MATERIAL AND METHODS: Sixty wbp x pbz sows after the first mating were slaughtered and tissues samples of the ovaries and oviduct were taken. Primer pairs for PCR analysis were designed on the basis of swine osteopontin 5' end sequence driven from GenBank. This study's amplified DNA fragment, spanning 274 bp of the promoter region, was chosen because of its contents of tree specific sites: type II collagen silence sequence (CACCTCC) at -682 (from the transcription initiation site), glucocortIcoid response site (TGTCCT) at -658 and CAAT box -592. This DNA fragment was subjected to a MSSCP analysis. RESULTS: A different MSSCP pattern was shown which indicates that mutation is located in this region. The samples of different conformers were sequenced and the A --> G transitions was identified in two positions -617 and -608. Restriction analysis of the DNA was performed, but unfortunately none of the known restriction enzymes recognized the novel SNPs, which is why the specific primer pairs characteristic for nucleotide A or for nucleotide G were chosen. In the second stage of the presented study the total RNA was extracted from the tissues of the ovaries and oviduct and the complementary DNA (cDNA) was synthesized. CONCLUSION: The real time PCR analysis to determined the expression dynamics of the OPN gene in pig tissues was performed. PMID- 20010487 TI - Solitary epileptic seizures in the clinical practice. Part II: findings of various modifications of EEG examination and imaging methods in patients who experienced solitary unprovoked epileptic seizure. AB - BACKGROUND: The occurrence of the first unprovoked epileptic seizure always requires thorough examination. The risk of incorrect diagnosis is high, non epileptic seizures represent 20-33% of newly diagnosed cases. The aim of the study is to evaluate findings of various modifications of EEG examination in the group of patients who experienced solitary unprovoked epileptic seizure and compare benefits of CT and MRI examination of the brain of these patients. MATERIAL AND METHODS: The group involved 84 patients hospitalized at the 1st Department of Neurology , Faculty of Medicine, Comenius University in Bratislava between January 1997 and January 2007 after experiencing unprovoked epileptic seizure. It is a retrospective analysis of information from medical records and clinical documentation. For statistic comparing of benefits of various modifications of EEG examination and between CT and MRI examination of the brain binomic test of proportion was used. RESULTS: Interictal epileptiform EEG graphoelements in native EEG was recorded only in 14.29% of patients. Statistically significantly more epileptiform graphoelements comparing EEG and EEG after sleep deprivation of the patient (SD) with one- hour recording was recorded with 24-hour 8 channels EEG monitoring after SD, EEG after SD with one- hour recording was of no significance comparing to native EEG. MRI examination of the brain was statistically more significant comparing to CT examination. CONCLUSION: Low catchment of epileptiform EEG graphoelements in patients with solitary unprovoked seizure shows the importance of precise history in diagnostics of these patients. Though EEG is a very important examination it is an auxiliary one. We confirmed that MRI examination in patients who experienced solitary unprovoked epileptic seizure is undoubtedly the first choice method. PMID- 20010488 TI - Solitary epileptic seizures in the clinical practice. Part I: etiological factors responsible for their occurrence. AB - BACKGROUND: Approximately 5% of the general population experiences at least one unprovoked epileptic seizure in the lifetime. This is in contrast to the cumulative incidence of epilepsy (approx. 3-4%) and incidence of acute symptomatic seizures (approx. 4%). Nearly 2% of the population experiences a febrile seizure before the 5 years of age. The aim of this article was evaluation of the distribution of acute symptomatic and unprovoked seizures in our patient cohort after the solitary epileptic seizures, as well as determination of particular etiological factors responsible for the occurrence of solitary epileptic seizure. MATERIAL AND METHODS: Our patient cohort comprised 116 patients experiencing the solitary epileptic seizure, who were hospitalised at the 1st Department of Neurology, Faculty of Medicine of Comenius University and Faculty Hospital in Bratislava, Slovakia over a period of 10 years (January 1, 1997 - January 1, 2007). It is a retrospective analysis of data from case records and health records; eventually we contacted the patients by phone or by sending a questionnaire. RESULTS: Of the 116 patients there were 32 cases (37.12%) of an acute symptomatic seizure and 84 cases (62.88%) of an unprovoked seizure. In the group of unprovoked solitary epileptic seizures the etiological conclusions were idiopathic and cryptogenic in 35 cases (41.7%) and late symptomatic in 49 cases (58.3%). The most frequent etiological factor in the group of acute symptomatic seizures was alcohol (40.6%). In the subgroup of late symptomatic seizures the etiology was mostly vascular (17.85%). CONCLUSION: Data management of etiological factors responsible for the solitary epileptic seizure confirmed a need for a thorough evaluation of the first epileptic seizure. PMID- 20010489 TI - Relationship between cytokine IL 6 levels and early-onset neonatal morbidity. AB - Perinatal infections exert considerably negative effects on morbidity and mortality of newborns. HYPOTHESIS: elevations of cytokine IL levels may be used as a marker of early-onset neonatal infections. TARGETS: to establish relationships between elevated IL 6 cytokine levels and neonatal morbidity (cranial and pulmonary). METHODS: examinations of the umbilical blood for IL 6 values in 392 newborns weighing under 1500 g treated in Ceske Budejovice at the Intensive Care Unit and Unit of Intensive Care and Resuscitation. RESULTS: a statistically significant relationship was demonstrated between elevations of cytokine IL 6 levels and pulmonary morbidity in newborns. DISCUSSION: the authors recommend examination of cytokine IL levels in the umbilical blood with a possibility to acquire the result within 2 hours after the delivery, which makes neonatologists possible to initiate goal-directed antibiotic therapy. PMID- 20010490 TI - Nonimmune hydrops fetalis. AB - BACKGROUND: Nonimmune hydrops fetalis (NHF) is an abnormal accumulation of fluid especially serous - in visceral cavities and soft tissues. This condition may be caused by: cardiovascular diseases, chromosomal disorders, infections, lung, stomach, intestinal, kidneys, urinary tract and blood diseases, metabolic disorders and tumors. NHF may by diagnosed by an ultrasound scan. THE AIM of the study was to present diagnostic and therapeutic difficulties as well as management with reference to NHF. CASE STUDY: An abnormal accumulation of fluid in visceral cavities and subcutaneous tissue of two fetuses was diagnosed by an ultrasound scan. Despite a detailed and specific diagnostic proceeding which included: infections, congenital malformations, chromosomal abnormalities etc. it was impossible to establish the cause of NHF. The symptomatic therapy was performed: periodic cordocentesis with an injection of human albumin solutions. In case of the first fetus therapeutic thoracocentesis was performed. The fetuses were delivered in 32nd and 31st week of pregnancy. Both neonates survived but even after the delivery it was impossible to establish the cause of NHF. CONCLUSIONS: Multidirectional diagnostic approach is essential for the implementation of causal treatment of NHF. In case of idiopathic NHF the only management is symptomatic therapy, fetal monitoring and preterm delivery. PMID- 20010491 TI - Neuroendocrine pathways altered in autism. Special role of reelin. AB - Autism is the most genetically influenced neuropsychiatric disorder with heritability of approximately 90%. Since genetic factors seem to play a crucial role in autism etiology, enormous attention is focused on genetic analyses of the disorder. Reelin, one of the autism candidates, is necessary in regulation of neuronal migration during brain development and also in maintaining synaptic plasticity during postnatal life period. Reduced reelin levels were observed in sera and brain cortices of autistic patients. In this review, abnormalities in reelin signaling and the relationship between reelin deficiency and principal neuroendocrine pathways are discussed. PMID- 20010492 TI - Neural correlates of the rejection of unfair offers in the impunity game. AB - OBJECTIVES: This study examined the roles of the insula and the anterior cingulate activations in the rejection of unfair offers in the impunity game. METHODS: Fifteen participants played the impunity game in ten trials as responders during neuroimaging. RESULTS: About 45% of the unfair offers were rejected by the responders even when responders could not restore a fair outcome, which cannot be accounted for by social preference of inequity aversion. Imaging data showed that the right anterior insula was activated when participants faced and rejected unfair offers. CONCLUSIONS: The insula activation during a rejection of the unfair offers is the reflection of an emotional response, rather than social preference of inequity aversion. The role of emotion in the neuroeconomics of fairness was demonstrated. PMID- 20010493 TI - Lower plasma Coenzyme Q10 in depression: a marker for treatment resistance and chronic fatigue in depression and a risk factor to cardiovascular disorder in that illness. AB - INTRODUCTION: There is now evidence that major depression is accompanied by an induction of inflammatory and oxidative and nitrosative stress (IO&NS) pathways and by a lowered antioxidant status. Coenzyme Q10 (CoQ10) is a strong antioxidant that has anti-inflammatory effects. METHODS: This paper examines the plasma concentrations of CoQ10 in 35 depressed patients and 22 normal volunteers and the relationships between plasma CoQ10 and treatment resistant depression (TRD), the severity of illness as measured by means of the Hamilton Depression Rating Scale (HDRS) and the presence of chronic fatigue syndrome (CFS). RESULTS: We found that plasma CoQ10 was significantly (p=0.0002) lower in depressed patients than in normal controls. 51.4% of the depressed patients had plasma CoQ10 values that were lower than the lowest plasma CoQ10 value detected in the controls. Plasma CoQ10 was significantly lower in patients with TRD and with CFS than in the other depressed patients. There were no significant correlations between plasma CoQ10 and the HDRS. DISCUSSION: The results show that lower CoQ10 plays a role in the pathophysiology of depression and in particular in TRD and CFS accompanying depression. It is suggested that depressed patients may benefit from CoQ10 supplementation. The findings that lower CoQ10 is a risk factor to coronary artery disease and chronic heart failure (CHF) and mortality due to CHF suggest that low CoQ10 is another factor explaining the risk to cardiovascular disorder in depression. Since statins significantly lower plasma CoQ10, depressed patients and in particular those with TRD and CFS represent populations at risk to statin treatment. PMID- 20010494 TI - Fear of falling has greater influence than other aspects of gait disorders on quality of life in patients with Parkinson's disease. AB - We assessed the influence of gait disorders (GD) on quality of life (QOL) in Parkinson's disease (PD). In a survey including the PDQ-39 and a new 8-item questionnaire of GD (8-QGD), 491 PD patients estimated their gait at the worst state, the effect of their GD on activities of daily living, compared the severity of GD with the other PD symptoms, evaluated the occurrence of freezing of gait (FOG), levodopa-resistant FOG, falls, activity limitation due to fear of falling (FOF), and the occurrence of injuries. Significant differences in PDQ-39 scores were found between three groups of patients divided with respect to the severity of GD. Linear multiple regression analysis showed that FOF had the highest impact on PDQ-39 scores (r=0.32, p<0.001). These results confirm that GD have a substantial impact upon the QOL in PD and suggest that FOF plays a major role in QOL deterioration. PMID- 20010495 TI - Effects of Iscador preparations on the reactivity of mouse immune system. AB - OBJECTIVES: Anticancer preparations made from plants have been an object of scientific interest for many years. It is worth noting that as many as 25% of cytostatics used in the anticancer chemotherapy are obtained from plants. One of the medical preparations which significantly influences cell metabolism is Iscador. Iscador preparations are used as complementary therapy in the conventional anticancer treatment. These are aqueous extracts of mistletoe (Viscum album L.). One repeatedly finds that mistletoe (Viscum album L.) extracts show immune-modulating effects. THE AIM at the present work was to study the influence of iscador Qu, M, P at a dose 5 mg/kg b.w., on the total protein concentration in blood serum and proportions of blood protein fractions determined by electrophoresis. Additionally leukocyte activity was estimated, which, served as indicators of the immune system reactivity in mice treated with anticancer preparations of vegetable origin. RESULTS: The experiment indicated statistically significant increase in albumin fraction level and lymphocyte count. Moreover, decrease of the total protein content, protein fractions globulins alpha2, beta, gamma and neutrophil, monocyte count in mouse serum was observed. PMID- 20010496 TI - Effectiveness of photodynamic therapy in the treatment of lichen sclerosus: cell changes in immunohistochemistry. AB - BACKGROUND: Vulvar lichen sclerosus (LS) affects primarily women at postmenopausal age and its background remains unknown. One of the treatment modalities is photodynamic therapy (PDT). The aim was to investigate the efficacy of PDT in women with LS and the analysis of protein expression before and after PDT. MATERIAL AND METHODS: From 04.2006-01.2008 28 women, with LS underwent photodynamic diagnosis and next PDT: six-courses every second week with using 5 aminolevulinic acid (ALA) as a photosensitizer. Punch biopsies were taken before and after treatment and immunohistochemistry was done with Ki67,CD44,CD34 and CD3. RESULTS: Before PDT all patients suffered from pruritus and after in 89.3% the relief was noted. The histological examination showed that 35.7% patients hadn't LS after therapy completion. Anti-CD44 staining intensities was scored qualitatively - there were no statistical difference at the expression of protein CD44 in the epidermis (p>0.05) before and after therapy. Microvessel density was assessed at the hot spots, marked with anti-CD34. Statistical difference in AVD before and after therapy: (p<0.05). The staining intensity of Ki-67 didn't differ before and after PDT (p>0.05). The expression of CD3 on T lymphocytes showed statistical difference of the lymphocytic infiltration before and after PDT ( p<0.05). CONCLUSION: The immunohistochemical staining in vulvar LS showed increasing microvessel density and decreasing lymphocytic infiltration. There were a clinical, and less histological improvement in patients with LS. We suggest that the photodynamic therapy is an effective, alternative treatment in some but not all patients with LS. Therefore, further studies are needed. PMID- 20010497 TI - Disrupted light-dark cycle induces obesity with hyperglycemia in genetically intact animals. AB - BACKGROUND: The environmental light-dark (LD) cycle entrains the circadian clock located in the suprachiasmatic nucleus (SCN) of mammals. Recent studies of genetically impaired animals with clock gene mutations have revealed associations between metabolic disorders and the circadian clock. However, whether such disordered phenotypes are due to a loss of circadian clock function within specific metabolically relevant tissues, or the result of disrupted circadian behavioral activities governed by the SCN remains unknown. OBJECTIVES: The present study examines the effect of disrupted LD cycles that might perturb the circadian clock in the SCN and peripheral organs on a high-fat/high-sucrose diet induced obesity in genetically intact mice. METHODS: The behavioral patterns of the mice were disturbed under an ultradian 3 h light-3 h dark cycle (LD 3:3) due to light-induced direct suppression of the behavior (masking effect). RESULTS: Obesity with hyperglycemia was significantly enhanced and levels of hemoglobin A1c were significantly higher under LD 3:3 compared with LD 12:12. CONCLUSIONS: These findings provide a link between metabolic disorders and the "environmental mutation" in genetically intact animals. PMID- 20010498 TI - Different nitrosative-induced microtubular modifications and testosterone neuroprotective effects on high-D-glucose-exposed neuroblastoma and glioma cells. AB - OBJECTIVES: Diabetic complications can often affect the central nervous system since the chronic exposure to hyperglycemia can result in the production of high concentration of reactive oxygen species with subsequent damage of several cell structures such as the cytoskeleton. In order to antagonize the oxidative status many substances have been tested as antioxidants. In the present work attention has been focused on the possible nitrosative effect of hyperglycemia on microtubular network of neuroblastoma and glioma mortalized cell lines, testing the possible neuroprotective effect of testosterone. METHODS: Neuroblastoma (C1300) and glioma (C6) cell lines were cultured in the presence of 300 mM (C1300) or 310 mM (C6) D-glucose, with or without 50 nM testosterone. After 72 hrs, morphology, growth rate, cell viability and catalase activity were evaluated. In addition, with the aim to detect any changes in the amount of tubulin isoforms, Western blot analysis was performed. RESULTS: In D-glucose exposed cells, it was found a down-regulation of tubulin isoforms and an increase in 3-nitro-L-tyrosine and subsequent modifications in cell morphology, growth rate, viability and catalase activity. All these changes were more severe in neuroblastoma than in glioma cell line. When testosterone was added to the medium, all the parameters were very similar to controls. This neuroprotective action was well-detectable in C1300 cells, whereas testosterone was not able to recover significantly in C6 cells. CONCLUSION: Our results displayed: i) a selective action of high glucose on microtubules; ii) a different sensitivity to oxidative stress in neuronal and glial cells; iii) a different neuroprotective action of testosterone on neuronal and glial cells. PMID- 20010499 TI - Different contribution of the lateral bed nucleus of the stria terminalis in intrahippocampal neostigmine-induced elevation of plasma glucose and adrenocorticotropic hormone in free moving rats. AB - OBJECTIVE: In our previous study, a lesion in the lateral bed nucleus of the stria terminalis (BNSTL) was found to significantly attenuate the elevation of adrenocorticotropic hormone (ACTH) in plasma during microinjectin of neostigmine, an inhibitor of acetylcholine esterase, into the rat hippocampus. The current study was designed to examine the role of the BNSTL in regulation of blood glucose elevation induced by hippocampal neostigmine injection. MATERIALS AND METHODS: Ibotenic acid (15 microg/microl) was stereotaxically bilaterally injected into the BNSTL of rats. Two weeks after the injections, neostigmine methylsulfate (sigma, 5 x 10-8 mol) was microinjected into the rat hippocampus in a volume of 1 microl for 1 min using a CMA/100 microinjection pump. Plasma ACTH and glucose concentrations were examined using radioimmunoassay and immobilized enzyme casing/H2O2 method techniques, respectively. RESULTS: Compared with sham operated control rats, rats with BNSTL lesions produced by ibotenic acid showed significantly attenuated the elevations of plasma ACTH evoked by the microinjection of neostigmine into the hippocampus. However, no significant difference of blood glucose in response to the injection was observed between the BNSTL-lesioned rats and controls. CONCLUSION: The results of the present study indicate that the BNSTL plays a role in ACTH regulation and not in blood glucose regulation when the hippocampal cholinergic system is activated. PMID- 20010500 TI - Diabetes mellitus decreases the expression of calcitonin-gene related peptide, gamma-amino butyric acid and glutamic acid decarboxylase in human pancreatic islet cells. AB - OBJECTIVES: The pattern of distribution of calcitonin-gene related peptide (CGRP), a neuropeptide, gamma-aminobutyric acid (GABA), a neurotransmitter and GABA-converting enzyme, glutamic acid decarboxylase (GAD) in the pancreas of diabetic patients was investigated to determine whether diabetes mellitus influences the expression of these biological transmitters. METHODS: Pancreatic tissue samples retrieved, during pancreatectomy, from cancer patients with and without Type 2 diabetes were paraffin embedded. The expression of CGRP, GABA and GAD was examined in pancreatic tissue using immunofluorescence techniques. RESULTS: CGRP, GABA and GAD were observed in many cells located in the central as well as the peripheral regions of pancreatic islet. The expression of CGRP, GABA and GAD decreased dramatically in pancreatic islet cells of diabetic patients compared to control. CGRP and GABA co-localized with glucagon in some pancreatic islet cells of both normal and diabetic patients. The pattern of distribution of CGRP, GABA and GAD in normal and Type 2 diabetic patients was similar to that of insulin. CONCLUSION: The number of human pancreatic islet cells expressing CGRP, GABA and GAD decreased significantly after the onset of Type 2 diabetes. These neuropeptides and neurotransmitters may play a role in the regulation of pancreatic beta cell function. PMID- 20010501 TI - Cytosine-Adenosine (CA)n repeats polymorphism in IGF-I gene and early growth in infants born appropriate and small for gestational age. AB - BACKGROUND: IGF-I gene polymorphisms might alter IGF-I level resulting in decreased foetal and postnatal growth and increased risk for diabetes mellitus type 2 and cardiovascular diseases in adulthood. OBJECTIVES: We analyzed the association between Cytosine-Adenosine (CA)10-24 repeats polymorphism in promoter region of the IGF-I gene and early growth in infants with birth weight appropriate for gestational age (AGA) and small for gestational age (SGA). DESIGN AND METHODS: All neonates were born at term, 196 of them were AGA and 26 SGA. Blood for DNA analyses was obtained from placental part of umbilical vein. Genotyping was performed using fragment analyses of IGF-I gene promoter region. The data about postnatal growth in the group of AGA children were obtained at the age of 18 months, in SGA children at 12 months. RESULTS: No differences in the frequency of wild type allele with (CA)19 repeats and polymorphisms with (CA)<19 or (CA)>19 repeats were observed between AGA and SGA children. The average birth weight and length in AGA wild type (CA)19 homozygotes were lower in comparison with AGA carriers of various (CA)n polymorphisms but all observed anthropometric differences disappeared at the age of 18 months. In SGA children, no differences were found between number of (CA)n repeats and anthropometric parameters both at birth and at the age of 12 months. CONCLUSIONS: Although (CA)n repeats polymorphism in IGF-I gene might affect prenatal growth in AGA children, our results have not shown any impact of variable number of (CA) n repeats in IGF-I gene on postnatal growth. PMID- 20010502 TI - Correlation of vitamin D receptor gene (VDR) polymorphism with osteoporotic changes in Polish postmenopausal women. AB - OBJECTIVE: Recently the significance of genetic traits, influencing hormonal and environmental factors, in susceptibility to osteopenia and osteoporosis development has been indicated. Much attention to the polymorphic variants of vitamin D receptor (VDR) gene was paid. The restriction polymorphisms in VDR gene could be involved in the modulation of vitamin D action and modulate the level of bone mineral density (BMD) and the risk to develop osteopenia and osteoporosis. MATERIALS AND METHODS: Total 321 postmenopausal women (mean age 63.26 +/- 8.90 years), including women with osteoporosis (163 patients) and osteopenia (95) have been compared to 63 women with normal t-score value. For detection of VDR polymorphisms PCR/RFLP (polymerase chain reaction/restriction fragment length polymorphism) assay have been used. RESULTS: The frequency of BsmI, ApaI, and TaqI polymorphic variants of VDR gene detected in investigated groups was not statistically different. The slight, not significant tendency to prevalence of a allele (ApaI polymorphism) in the controls comparing to women with osteoporosis and osteopenia have been noted. Higher prevalence of homozygous TT genotype (TaqI polymorphism) the in the both groups with lower BMD value (47.9 : 49.5 vs. 34.9% in the controls) and higher prevalence of T allele in these both groups (65.9 : 68.4 vs. 57.9) was been also observed. CONCLUSIONS: The presence of T allele of TaqI polymorphism could predict the higher risk to develop osteoporosis in postmenopausal woman; consequently t allele could have protective effect. The presence of A allele (ApaI polymprphism) seems to be weakly connected with osteoporosis susceptibility. PMID- 20010503 TI - Complete objective response of neuroblastoma to biological treatment. AB - OBJECTIVES: The combined use (MDB) of Somatostatin, Melatonin, Retinoids, Vitamins E, C, and D3, with Calcium, Chondroitin sulfate, and microdoses of Cyclophosphamide in a seven-month old baby affected by a voluminous retroperitoneal neuroblastoma measuring 4 x 8 cm produced a 50% objective response in six months, an almost total response in one year and a complete response at 14 months, with cure and absence of disease for over ten years. RESULTS: This paper discusses the rationale and the molecular mechanisms of action of the treatment which has a differentiating, apoptotic and antiproliferative effect, preserving and enhancing both the trophism and functionality of organs and tissues, and the neuroimmunoendocrine and antiblastic homeostasis. This result is in agreement with the positive results already published on the use of the MDB in lymphoproliferative diseases, in stage III and IV lung cancer, in breast cancer and in cancers of the upper aerodigestive epithelia. Without the need for hospitalisation and with no toxicity, the MDB rapidly reduced and then eliminated the voluminous tumoral mass, allowing a normal quality of life and a perfectly normal psychophysical development. CONCLUSIONS: We believe it is of use to report this case in order to invite greater interest in the oncological possibilities offered by the immunoneuroendocrine and biological-receptorial properties of the MDB treatment. PMID- 20010504 TI - Coexpression of survivin and PCNA in pituitary tumors and normal pituitary. AB - OBJECTIVES: The survivin is the protein involved in regulation of basic and cycle specific functions of cells both in normal and cancer tissue. Recent studies present survivin as a factor having the leading role in the regulation of apoptosis and mitosis as well as a target of anticancer therapy. The employing of survivin in this therapy is based on its high expression level in most human cancers, as well as its association with the disease's progression. The aim of our study was to evaluate the expression and localization of survivin's gene product on the protein level in different types of pituitary tumors and normal pituitary. The coexpression of survivin and proliferating cell nuclear antigen - PCNA in pituitary was also examined. DESIGN AND METHODS: The study was conducted on the postoperative pituitary tumors tissue taken during standard neurosurgical removal of tumor from 43 patients. The group of patients consists of 23 women and 20 men, aged from 27 to 71 years. As a control of the study three normal pituitary tissues obtained at the autopsy were used. Evaluation of survivin and PCNA expression was performed using immunohistochemical staining. RESULTS: The study demonstrated the presence of survivin in all analyzed by us pituitary tumors. Survivin was present also in normal pituitary tissue. The protein was localized mainly in cell's nuclei, however the less intense immunostaining was observed also in the cytoplasm of pituitary tumors cells. Furthermore survivin was found in normal pituitary, but the positive immunostaining was limited to a single cells. The analysis of pituitary tumor cells proliferation index based on PCNA reactivity showed that survivin is coexpressed with PCNA, especially in invasive tumors. CONCLUSIONS: The study documented the presence of survivin in different types of pituitary tumors as well as in normal pituitary. Additionally the coexpression of survivin and PCNA in tumor cells was shown. The expression of survivin in both normal and cancer pituitary cells suggests that it may play an important role in regulation of the gland's proliferation. PMID- 20010505 TI - Coenzyme Q10 deficiency in myalgic encephalomyelitis/chronic fatigue syndrome (ME/CFS) is related to fatigue, autonomic and neurocognitive symptoms and is another risk factor explaining the early mortality in ME/CFS due to cardiovascular disorder. AB - INTRODUCTION: Myalgic encephalomyelitis/chronic fatigue syndrome (ME/CFS) is a medical illness characterized by disorders in inflammatory and oxidative and nitrosative (IO&NS) pathways. METHODS: This paper examines the role of Coenzyme Q10 (CoQ10), a mitochondrial nutrient which acts as an essential cofactor for the production of ATP in mitochondria and which displays significant antioxidant activities. Plasma CoQ10 has been assayed in 58 patients with ME/CFS and in 22 normal controls; the relationships between CoQ10 and the severity of ME/CFS as measured by means of the FibroFatigue (FF) scale were measured. RESULTS: Plasma CoQ10 was significantly (p=0.00001) lower in ME/CFS patients than in normal controls. Up to 44.8% of patients with ME/CFS had values beneath the lowest plasma CoQ10 value detected in the normal controls, i.e. 490 microg/L. In ME/CFS, there were significant and inverse relationships between CoQ10 and the total score on the FF scale, fatigue and autonomic symptoms. Patients with very low CoQ10 (<390 microg/L) suffered significantly more from concentration and memory disturbances. DISCUSSION: The results show that lowered levels of CoQ10 play a role in the pathophysiology of ME/CFS and that symptoms, such as fatigue, and autonomic and neurocognitive symptoms may be caused by CoQ10 depletion. Our results suggest that patients with ME/CFS would benefit from CoQ10 supplementation in order to normalize the low CoQ10 syndrome and the IO&NS disorders. The findings that lower CoQ10 is an independent predictor of chronic heart failure (CHF) and mortality due to CHF may explain previous reports that the mean age of ME/CFS patients dying from CHF is 25 years younger than the age of those dying from CHF in the general population. Since statins significantly decrease plasma CoQ10, ME/CFS should be regarded as a relative contraindication for treatment with statins without CoQ10 supplementation. PMID- 20010506 TI - Cell signalling in CNS and immune system in depression and during antidepressant treatment: focus on glial and natural killer cells. AB - CNS, endocrine and immune systems share the same molecules: neurotransmitters, cytokines and hormones to communicate within and among each other. Depression is associated with abnormalities in the noradrenergic, serotonergic and dopaminergic neurotransmitter systems and reductions in the level of their precursors and metabolic turnover. Most of these signalling molecules use trimeric G-proteins as a transduction system to transfer extracellular signal into cellular response. Altered levels or function of signalling proteins, especially alpha subunits of trimeric G-proteins, were found in post-mortem brain tissue and leukocytes of subject suffering from major depression. There is a considerable evidence that inflammatory response and immune system changes are the part of depression. Components of cellular immune system natural killer cells, important effectors of immune surveillance, are sensitive to stress response, and their functions are compromised in depressive subjects. Many lines of evidence also point to the loss of both neuronal and glial plasticity and neurotrophic factor support under chronic stress or in depression. There is an increasing knowledge of the role of astrocytic cells in neuroplastic processes and neurotransmitter metabolism. Alterations in the glial populations are observed in major depressive subjects. Antidepressant treatment is modulating glial signalization cascades, increasing production of neurotrophic molecules, supporting neuroplasticity processes, and also modulating functions of natural killers. At the level of membrane signalling, antidepressants show a direct influence upon G alpha subunit levels in both immune system and CNS. These findings support the view that antidepressants influence activity of natural killer and astrocytic populations, and this could be of importance in the depression etiopathogenesis and/or treatment. PMID- 20010507 TI - Baroreflex sensitivity in patients with type I diabetes mellitus. AB - OBJECTIVES: To date, the clinical usefulness of measuring baroreflex sensitivity (BRS) to detect impairment of the autonomic nervous system in patients with diabetes mellitus (DM) type I has not been evaluated sufficiently (Mlcakova et al. 2008). The aim of the current study was the determination and statistical comparison of the mean values of BRS in our DM type I patients cohort and in a control group of healthy volunteers as well as the determination of BRS value dependency on the duration of diabetes and the level of glycemic control in DM I patients. We also aimed to determine the inter-individual and intra-individual variability of BRS in our patients. MATERIAL AND METHODS: We examined 100 patients with type I diabetes mellitus (37 women and 63 men, mean age 30 years, duration of the disease >or= 10 years) and 40 healthy, age- and sex-matched, subjects. Data from the patient cohort were subsequently analysed for duration of the diabetes and the level of glycemic control as assessed by glycated haemoglobin (HbA1c). We used a simple proportional test to compare the occurrence of impaired BRS in the patient cohort and the control group, and a simple linear regression to assess associations between BRS and duration of the diabetes and the levels of glycemic control. RESULTS: The mean BRS value in our group of diabetic patients and the control group were 10.15 ms/mmHg and 13.35 ms/mmHg, respectively. II. Statistically significant association between BRS impairment and the duration of the disease or level of glycemic control was not confirmed in our patient cohort. III. We observed an increased inter-individual variability and a relatively low intra-individual variability of BRS in patients with DM type I. CONCLUSIONS: We found a statistically highly significant difference between the proportions of impaired BRS in the group of diabetics vs. control. However, BRS did not correlate with the duration of the disease or with the level of glycemic control significantly. Albeit not reaching statistical significance, trends could be observed, which we consider clinically interesting. PMID- 20010508 TI - Deaths related to 2009 pandemic influenza A (H1N1) among American Indian/Alaska Natives - 12 states, 2009. AB - Indigenous populations from Australia, Canada, and New Zealand have been found to have a three to eight times higher rate of hospitalization and death associated with infection with the 2009 pandemic influenza A (H1N1) virus. In October, two U.S. states (Arizona and New Mexico) observed a disproportionate number of deaths related to H1N1 among American Indian/Alaska Natives (AI/ANs). These observations, plus incomplete reporting of race/ethnicity at the national level, led to formation of a multidisciplinary workgroup comprised of representatives from 12 state health departments, the Council of State and Territorial Epidemiologists, tribal epidemiology centers, the Indian Health Service, and CDC. The workgroup assessed the burden of H1N1 influenza deaths in the AI/AN population by compiling surveillance data from the states and comparing death rates. The results indicated that, during April 15-November 13, AI/ANs in the 12 participating states had an H1N1 mortality rate four times higher than persons in all other racial/ethnic populations combined. Reasons for this disparity in death rates are unknown and need further investigation; however, they might include a high prevalence of chronic health conditions (e.g., diabetes and asthma) among AI/ANs that predisposes them to influenza complications, poverty (e.g., poor living conditions), and delayed access to care. Efforts are needed to increase awareness among AI/ANs and their health-care providers of the potential severity of influenza and current recommendations regarding the timely use of antiviral medications. Efforts to promote the use of 2009 H1N1 influenza monovalent vaccine in AI/AN populations should be expanded. PMID- 20010509 TI - Fatal poisoning among young children from diethylene glycol-contaminated acetaminophen - Nigeria, 2008-2009. AB - On November 18, 2008, the Nigerian Federal Ministry of Health (FMOH) received a report of 13 cases of unexplained acute renal failure among children from a hospital in Lagos state. Several of the patients had been exposed to a liquid acetaminophen-based teething medication. On November 21, officials from the Nigerian National Agency for Food and Drug Administration and Control (NAFDAC) discovered diethylene glycol (DEG) in four batches of the teething medication manufactured during August-October 2009. DEG is a toxic alcohol used in brake fluid, paint, and household cleaning products, and has been used illegally as a cheap substitute solvent in drug manufacturing. Previous DEG poisonings resulting from contamination of medications have been reported in the United States, Nigeria (1990), Panama, and other countries, and acute renal failure (ARF) is a known manifestation of DEG poisoning. An investigation was launched by the Nigeria Field Epidemiology and Laboratory Training Program (N-FELTP), CDC, and the Food and Drug Administration (FDA). This report summarizes the results of the investigation, which identified 57 cases of DEG poisoning among children aged 5 and < 8 (n = 8); group 3 - with the highest Gleason scale > 8 (n = 6). RESULTS: Upon histological examination of prostate biopsy specimens, it was found that in the first group in 6 out of 13 (46%) cases small acinic cell PC developed on the background of chronic prostatitis with PIA (proliferative inflammatory atrophy) locus, frequently in combination with prostatic intraepithelial neoplasia (PI) locus. Hyperchromic epithelial cells in PIA locus were characterized by nuclear expression of p53 and Ki67 proteins, and cytoplasmic expression of Bcl-2. The precancerous foci in the PIN and PIA in the biopsy specimens of the second group of PC patients were found in 2 out of 8 (25%) cases of large and small acinic cell adenocarcinoma observations. The expression level of p53, p16INK4a, Bcl-2 proteins and especially Ki67 protein adequately increased in tumors of group 2 in comparison with group 1. Group 3 comprised of patients with Gleason scale > 8, predominantly solid structures or scirrhus of PC, which were characterized by the highest nuclear expression of p53, p16INK4a and Ki-67, and also by overexpression of cytoplasmic Bcl-2. CONCLUSIONS: Obtained results showed the direct correlation between patients' Gleason scale, and the expression level of p53, p16INK4a, Bcl-2 proteins and, particularly, Ki67 marker of proliferating cells in PC tumor cells. PMID- 20010527 TI - Expression of E-cadherin in drug resistant human breast cancer cells and their sensitivity to lymphokine-activated lymphocytes action. AB - AIM: To analyze the correlation between the elevated sensitivity of drug resistant breast cancer cells to the action of lymphokine-activated lymphocytes (LAK) and expression of E-cadherin and other marker proteins by cancer cells and lymphocytes. METHODS: Breast tumor explants were cultured with autologous lymphocytes in double diffusion chambers. The results were evaluated by morphological criteria of explants growth. Expression level of proteins on tumor cells was analyzed using immunohistochemical method on paraffin embedded sections, and by indirect immunofluorescence - on lymphocytes. RESULTS: Significant decrease of E-cadherin expression and significant increase of nuclear antigen of proliferating cells expression have been detected on drug resistant malignant human breast tumor (DRHBT) cells compared with drug sensitive breast tumor (DSHBT) cells. Autologous LAK possessed the highest antitumor activity against DRHBT cells that was associated with high expression level of soybean lectin receptor. CONCLUSION: Malignant drug resistant tumors are characterized by reverse relation between E-cadherin expression level and their proliferative activity. Marked antitumor action of LAK against these tumors is associated with high expression level of soybean lectin receptor on the lymphocytes. PMID- 20010528 TI - The prognostic value of cytogenetic markers for early diagnosis of colorectal cancer. AB - AIM: To investigate the spectrum of chromosome changes in colorectal adenomas and adenocarcinomas and to evaluate the prognostic significance of the chromosome rearrangements. METHODS: The study was carried out using the cytogenetic analysis of biopsy specimens (n = 56) of single and multiply adenomas (familial adenomatous syndromes; n = 38) and adenocarcinomas (n = 18). RESULTS: The karyotype of adenomas was normal in the majority of cases, but some adenomas with severe dysplasia of epithelium carry the quantitative chromosome abnormalities and structural rearrangements. The combination of additional copies of chromosomes 13, 18, 20 in adenomas points on an unfavorable prognosis. The chromosome abnormalities were found in 100% of adenocarcinomas biopsy specimens. CONCLUSIONS: The transition from colorectal adenomas to adenocarcinomas is accompanied by elevation of chromosome abnormalities level, in particular, by increased clonal variety, selective accumulation of the copies of chromosomes 2, 3, 20, 16, with simultaneous monosomy of chromosomes 17, 18, 8, 6, 14, and del 1p. PMID- 20010529 TI - Relation between pathomorphological response in tumors after neoadjuvant chemotherapy and clinico-morphological and molecular prognostic factors in patients with breast cancer. AB - AIM: To determine the correlation between tumor pathomorphological response (PMR) after neoadjuvant chemotherapy (NACT) and clinico-morphological and molecular prognostic factors in patients with breast cancer (BC), and to determine the possible impact of the PMR and estrogen receptors (ER), progesterone receptors (PR) and Her-2/neu BC status on the disease course. METHODS: The data from the medical history of patients on IIB stage (T2N1M0, T3N0M0) (n = 247), who received treatment with NACT, were used. The correlation between the parameters was determined using the Spearman's coefficient. Patient's survival was analyzed by Kaplan - Meier method. The association between PMR grades with the risk of disease relapse was estimated by Cox's regression analysis. RESULTS: PMR grade correlated with tumor differentiation grade (rho = 0.38; p 0.05) and BC subtypes (rho = 0.05; p > 0.05). The patients with the same PMR grades didn't differ by the number of lymph node metastases (p > 0.05) and differed by the presence of embolus in tumor vessels (p 0.05). The risk of disease relapse depended on PMR grade: for grade 2-3 it was significantly decreased (HR = 0.71, 95% CI - 0.25 2.9, p = 0.0037), and for grade 4-5 it was the highest (HR = -1.23, 95% CI - 0.24 5.05, P = 0.0001), while 0-1 grade had no impact on the risk of disease relapse (HR = 0.22, 95% CI 0.08-0.38; p = 0.7). CONCLUSION: The data of combined clinical, histological and immunohistochemical analysis have shown that PMR grades may serve as the criteria for individualization of adjuvant treatment of the patients with locally advanced BC. PMID- 20010530 TI - Antitumor and antimetastatic activities of vaccine prepared from cisplatin resistant lewis lung carcinoma. AB - AIM: To study antitumor and antimetastatic activities of antitumor vaccine (ATV) prepared from cisplatin (CP) sensitive and resistant strains of Lewis lung carcinoma (LLC). METHODS: The inhibition of tumor growth, and the mean survival time of the tumor-bearing animals, the number and the volume of metastases were measured as the indices of ATV efficacy. The activity of cytotoxic T-lymphocytes and natural killer cells, peritoneal macrophages (Mph), the level of tumor necrosis factor and the total proteolytic activity of blood plasma (PA) were assessed. RESULTS: ATV from CP resistant LLC prepared using cytolectin (CL) of capital VE, Cyrillic. subtilis capital VE, Cyrillic-7025 significantly inhibited growth of CP resistant tumors (by 52%) and increased mean survival time (MST) of animals (by 44.6%). The index of metastasis inhibition for ATV prepared from CP sensitive or resistant LLC was 154.5% and 227.0%, respectively. In all vaccine treated animals, Mph activity was shown to be significantly increased. In spite of high antitumor and antimetastatic effects of ATV prepared from CP resistant LLC, PA in plasma of animals inoculated with CP resistant LLC was increased significantly upon vaccine administration. PMID- 20010531 TI - The effect of racemic gossypol and at-101 on angiogenic profile of ovcar-3 cells: a preliminary molecular framework for gossypol enantiomers. AB - AIM: To compare the effect of racemic gossypol with its (-)/(-) enantiomer (AT 101) on expression profiles of angiogenic molecules by mRNA levels in human ovarian cancer cell line OVCAR-3. METHODS: Cell viability assay (2,3-bis (2 methoxy-4-nitro-5- sulfophenyl)-5-[(phenylamino) carbonyl]-2H-tetrazolium hydroxide) was used to detect cytotoxicity of gossypol enantiomers. DNA fragmentation by an enzyme-linked immunosorbent (ELISA) assay was used to evaluate the rate of apoptosis. The mRNA expression levels of angiogenic molecules were investigated by Human Angiogenesis RT2 ProfilerTM PCR Array (SuperArray, Frederick, MD). RESULTS: Both racemic form and AT-101 resulted in a significant cytotoxicity and induced apoptosis. This effect was observed in a dose- and time dependent manner. However, AT-101 was much more potent. In addition, the treatment of 10 microM of racemic gossypol alone and 3 microM of AT 101 alone resulted in significant down-regulation (>or= 3 fold) in mRNA levels of some pivotal angiogenic molecules in OVCAR-3, but altered gene profiles were different by the treatment of each enantiomer. CONCLUSION: The efficacy of two gossypol enantiomers in OVCAR-3 cells showed distinction. AT-101 was much more potent than racemic gossypol, not only by means of cell death and apoptosis, but also by modulation of angiogenic molecules released from OVCAR-3 cells. Further studies with endothelial cells should be done to verify the anti-angiogenic effect of gossypol enantiomers in cancer treatment. PMID- 20010532 TI - Antiangiogenic properties of a nutrient mixture in a model of hemangioma. AB - The pathogenesis of hemangiomas is still largely unknown and the current therapy, such as systemic corticosteroid, vincristine, and interferon-alpha, is toxic and remains unsatisfactory. A nutrient mixture (NM) containing lysine, proline, ascorbic acid and green tea extract has shown significant anti-angiogenic and anti-tumor effect against a number of cancer cell lines. AIM: Using a mouse hemangioendothelioma model, we investigated the efficacy of NM. We also tested the effect of NM in vitro, evaluating cell viability, MMP secretion, invasion, morphology and apoptosis. METHODS: Athymic nude mice, 5-6 weeks old, were inoculated with 3 x10(6) EOMA cells subcutaneously and randomly divided into two groups; group A was fed a regular diet and group B - a regular diet supplemented with 0.5% NM. Four weeks later, the mice were sacrificed and their tumors were excised, weighed and processed for histology. We also tested the effect of NM in vitro. RESULTS: NM inhibited the growth of tumors by 50%. In vitro, NM exhibited dose response cytotoxicity with 10%, 30% and 55% at 10, 100 and 1000 microg/ml. Invasion through Matrigel was inhibited at 50, 100 and 500 microg/ml by 25%, 30% and 100% respectively. NM induced dose-dependent apoptosis of EOMA cells. CONCLUSIONS: These results suggest that NM may have therapeutic potential in treating infantile hemangioendotheliomas and, perhaps, other cutaneous vascular tumors. PMID- 20010533 TI - Serum factors that suppress cytotoxic effect of methotrexate. AB - AIM: To study the phenomenon that human erythroid leukemia K-562 cells are more sensitive to cytotoxic effect of antimetabolites when cultured in a serum-free medium than in a conventional medium containing fetal calf serum (FCS). METHODS: Cytotoxic effects of methotrexate, azaserine and 5-fluorouracil were estimated by accessing the lactate dehydrogenase (LDH) activity of viable tumor cells. Proteins of FCS were separated using two-dimensional electrophoresis followed by mass spectrometry analysis. RESULTS: Addition of 10% FCS attenuated anti-tumor activity of methotrexate and azaserine against K-562 cells compared with serum free medium. Such an activity of FCS was different for each serum lot. Comparison of the proteins in active serum lot with those in not active one using two dimensional electrophoresis showed that in the active serum there were proteins 150 kDa, which were absent in the not active serum lot. Mass spectrometry indicated that all those proteins had the amino acid sequence of albumin. Sera of one healthy volunteer and two patients with thyroid cancer also attenuated the activity of the agent. CONCLUSION: Several lots of FCS and human serum demonstrated the ability to attenuate the cytotoxic effect of methotrexate in vitro, possibly due to the formation of albumin dimers/MTX complexes. PMID- 20010534 TI - The role of ultraviolet radiation and tyrosine stimulated melanogenesis in the induction of oxidative stress alterations in fair skin melanocytes. AB - BACKGROUND: Melanocytes are producing melanin after UV irradiation as a defense mechanism. However, UV-induced damage is involved in melanoma initiation, depending on skin phototype. Melanocytes seem to be extremely susceptible to free radicals. Their main enzymatic antioxidants are superoxide dismutase and catalase. AIM: To study how melanin synthesis modulates the activity of the oxidative stress defense enzymes and cell proliferation after UV induced cell damage. METHODS: Normal human melanocyte cultures from fair skin individuals were exposed to high levels of L-tyrosine and irradiated, with 20, 30, 40 mJ/cm2 UVA, and respective UVB. Proliferation was measured using a MTS assay; viability was assessed by trypan blue exclusion dye method. Spectrophotometrical methods were used to determine total melanin content, the enzymatic activity of tyrosinase, superoxide dismutase and catalase. RESULTS: Tyrosine had a negative effect on proliferation, enhanced with time elapsed. Overall, UV irradiation decreased proliferation. UVA increased proliferation relative to UVB in the cultures exposed for a longer time to high (2 mM) tyrosine concentration. There were no proliferation differences between UVA and UVB irradiation in lower tyrosine concentration exposed melanocytes. Both, UV irradiation and tyrosine increased melanogenesis. Exposure of the melanocytes to increased levels of tyrosine in medium (0.5 mM and 1 mM) and UV irradiation enhanced the activity of superoxide dismutase and catalase. The enzymes showed a high activity rate in melanocytes while exposed for a short time to 2 mM tyrosine, but their activity was dramatically decreased with longer tyrosine exposure and UV irradiation. CONCLUSION: Our data indicate that in low phototype melanocytes, melanogenesis, either following UV irradiation, or tyrosine exposure, especially in high concentrations, was detrimental for the cells by reducing the activity of catalase and superoxidedismutase, the natural antioxidants. UVA was more efficient in stimulating the activity of superoxide dismutase and catalase but also in depleting the reserves of the enzymatic defense against oxidative stress, especially catalase, than UVB. This physiologic response to UV light can be considered as an adjunctive risk factor for people with low phototype for developing a melanoma, when exposed to UV irradiation. PMID- 20010535 TI - Photo-oxidative action in cervix carcinoma cells induced by HPD - mediated photodynamic therapy. AB - Photodynamic therapy leads to oxidative stress through the generation of free radicals. Oxidative stress causes damage to cellular macromolecules such as nucleic acids, proteins and lipids. AIM: To examine the hematoporphyrin derivative (HpD) - mediated photodynamic effect on cervical adenocarcinoma cell line HeLa. METHODS: The HpD localization in HeLa cells was analyzed by confocal microscopy with epi-fluorescence system. Lipid peroxidation (LPO) was estimated by measurement of the concentration of malondialdehyde, protein degradation - by modified Ellman's method, superoxide dysmutase (SOD) - using Ransod Kit. The expression of inducible nitric oxide synthase (iNOS) was detected by immunocytochemical staining. RESULTS: The HpD was distributed all over the cytoplasm with preferential localization in the inner side of the plasma membrane and around the nuclear envelope. The process of photosensitizer distribution was time dependent. PDT-HpD increased the level of malonodialdehyde (MDA), SOD activity and the expression of iNOS in HeLa cells. However, PDT induced the decrease in the level of protein-associated thiol groups. CONCLUSIONS: Our study showed the important role of PDT-mediated oxidative stress in HeLa cells. HpD-PDT might be alternative and less invasive approach for treatment of patients with cervical cancer resistant for standard chemotherapy and radiotherapy. PMID- 20010536 TI - Are monosomies underestimated while analyzing cancer cells? PMID- 20010537 TI - Devotion to science ideals (to 110 th birthday anniversary of academician R.E. Kavetsky). PMID- 20010538 TI - Inflammation and cancer. PMID- 20010539 TI - The 2009 Lindau Nobel Laureate Meeting: Peter Agre, Chemistry 2003. AB - Peter Agre, born in 1949 in Northfield Minnesota, shared the 2003 Nobel Prize in Chemistry with Roderick MacKinnon for his discovery of aquaporins, the channel proteins that allow water to cross the cell membrane. Agre's interest medicine was inspired by the humanitarian efforts of the Medical Missionary program run by the Norwegians of his home community in Minnesota. Hoping to provide new treatments for diseases affecting the poor, he joined a cholera laboratory during medical school at Johns Hopkins. He found that he enjoyed biomedical research, and continued his laboratory studies for an additional year after medical school. Agre completed his clinical training at Case Western Hospitals of Cleveland and the University of North Carolina, and returned to Johns Hopkins in 1981. There, his serendipitous discovery of aquaporins was made while pursuing the identity of the Rhesus (Rh) antigen. For a century, physiologists and biophysicists had been trying to understand the mechanism by which fluid passed across the cell's plasma membrane. Biophysical evidence indicated a limit to passive diffusion of water, suggesting the existence of another mechanism for water transport across the membrane. The putative "water channel," however, could not be identified. In 1988, while attempting to purify the 30 kDa Rh protein, Agre and colleagues began investigating a 28 kDa contaminant that they believed to be a proteolytic fragment of the Rh protein. Subsequent studies over the next 3-4 years revealed that the contaminant was a membrane-spanning oligomeric protein, unrelated to the Rh antigen, and that it was highly abundant in renal tubules and red blood cells. Still, they could not assign a function to it. The breakthrough came following a visit with his friend and former mentor John Parker. After Agre described the properties of the mysterious 28 kDa protein, Parker suggested that it might be the long-sought-after water channel. Agre and colleagues tested this idea by expressing the protein in Xenopus oocytes, which typically have low water permeability. When the test oocytes were placed in a hypotonic solution, they swelled and exploded, thus revealing the function of the unknown protein as a water channel, which they named aquaporin. The Nobel Prize enabled Agre to take his research and scientific interests in new directions. He felt that over the years his work had continually taken him further from his original interests in third-world diseases, so he shifted his focus back in that direction. He now serves as the director of the Malaria institute at Johns Hopkins where he has applied his knowledge to the study of the malarial parasite and the Anopheles mosquito, which both express aquaporins. In addition, since winning the Nobel Prize, he has enjoyed increased opportunities for bringing science to the public and for "encouraging young people to go into science." PMID- 20010540 TI - Intranuclear microinjection of DNA into dissociated adult mammalian neurons. AB - Primary neuronal cell cultures are valuable tools to study protein function since they represent a more biologically relevant system compared to immortalized cell lines. However, the post-mitotic nature of primary neurons prevents effective heterologous protein expression using common procedures such as electroporation or chemically-mediated transfection. Thus, other techniques must be employed in order to effectively express proteins in these non-dividing cells. In this article, we describe the steps required to perform intranuclear injections of cDNA constructs into dissociated adult sympathetic neurons. This technique, which has been applied to different types of neurons, can successfully induce heterologous protein expression. The equipment essential for the microinjection procedure includes an inverted microscope to visualize cells, a glass injection pipet filled with cDNA solution that is connected to a N2(g) pressure delivery system, and a micromanipulator. The micromanipulator coordinates the injection motion of microinjection pipet with a brief pulse of pressurized N2 to eject cDNA solution from the pipet tip. This technique does not have the toxicity associated with many other transfection methods and enables multiple DNA constructs to be expressed at a consistent ratio. The low number of injected cells makes the microinjection procedure well suited for single cell studies such as electrophysiological recordings and optical imaging, but may not be ideal for biochemical assays that require a larger number of cells and higher transfection efficiencies. Although intranuclear microinjections require an investment of equipment and time, the ability to achieve high levels of heterologous protein expression in a physiologically relevant environment makes this technique a very useful tool to investigate protein function. PMID- 20010541 TI - Osmotic avoidance in Caenorhabditis elegans: synaptic function of two genes, orthologues of human NRXN1 and NLGN1, as candidates for autism. AB - Neurexins and neuroligins are cell adhesion molecules present in excitatory and inhibitory synapses, and they are required for correct neuron network function. These proteins are found at the presynaptic and postsynaptic membranes. Studies in mice indicate that neurexins and neurologins have an essential role in synaptic transmission. Recent reports have shown that altered neuronal connections during the development of the human nervous system could constitute the basis of the etiology of numerous cases of autism spectrum disorders. Caenorhabditis elegans could be used as an experimental tool to facilitate the study of the functioning of synaptic components, because of its simplicity for laboratory experimentation, and given that its nervous system and synaptic wiring has been fully characterized. In C. elegans nrx-1 and nlg-1 genes are orthologous to human NRXN1 and NLGN1 genes which encode alpha-neurexin-1 and neuroligin-1 proteins, respectively. In humans and nematodes, the organization of neurexins and neuroligins is similar in respect to functional domains. The head of the nematode contains the amphid, a sensory organ of the nematode, which mediates responses to different stimuli, including osmotic strength. The amphid is made of 12 sensory bipolar neurons with ciliated dendrites and one presynaptic terminal axon. Two of these neurons, named ASHR and ASHL are particularly important in osmotic sensory function, detecting water-soluble repellents with high osmotic strength. The dendrites of these two neurons lengthen to the tip of the mouth and the axons extend to the nerve ring, where they make synaptic connections with other neurons determining the behavioral response. To evaluate the implications of neurexin and neuroligin in high osmotic strength avoidance, we show the different response of C. elegans mutants defective in nrx-1 and nlg-1 genes, using a method based on a 4M fructose ring. The behavioral phenotypes were confirmed using specific RNAi clones. In C. elegans, the dsRNA required to trigger RNAi can be administered by feeding. The delivery of dsRNA through food induces the RNAi interference of the gene of interest thus allowing the identification of genetic components and network pathways. PMID- 20010542 TI - Lensless on-chip imaging of cells provides a new tool for high-throughput cell biology and medical diagnostics. AB - Conventional optical microscopes image cells by use of objective lenses that work together with other lenses and optical components. While quite effective, this classical approach has certain limitations for miniaturization of the imaging platform to make it compatible with the advanced state of the art in microfluidics. In this report, we introduce experimental details of a lensless on chip imaging concept termed LUCAS (Lensless Ultra-wide field-of-view Cell monitoring Array platform based on Shadow imaging) that does not require any microscope objectives or other bulky optical components to image a heterogeneous cell solution over an ultra-wide field of view that can span as large as approximately 18 cm(2). Moreover, unlike conventional microscopes, LUCAS can image a heterogeneous cell solution of interest over a depth-of-field of approximately 5 mm without the need for refocusing which corresponds to up to approximately 9 mL sample volume. This imaging platform records the shadows (i.e., lensless digital holograms) of each cell of interest within its field of view, and automated digital processing of these cell shadows can determine the type, the count and the relative positions of cells within the solution. Because it does not require any bulky optical components or mechanical scanning stages it offers a significantly miniaturized platform that at the same time reduces the cost, which is quite important for especially point of care diagnostic tools. Furthermore, the imaging throughput of this platform is orders of magnitude better than conventional optical microscopes, which could be exceedingly valuable for high-throughput cell-biology experiments. PMID- 20010543 TI - A novel RFP reporter to aid in the visualization of the eye imaginal disc in Drosophila. AB - The Drosophila eye is a powerful model system for studying areas such as neurogenesis, signal transduction and neurodegeneration. Many of the discoveries made using this system have taken advantage of the spatiotemporal nature of photoreceptor differentiation in the developing eye imaginal disc. To use this system it is first necessary for the researcher to learn to identify and dissect the eye disc. We describe a novel RFP reporter to aid in the identification of the eye disc and the visualization of specific cell types in the developing eye. We detail a methodology for dissection of the eye imaginal disc from third instar larvae and describe how the eye-RFP reporter can aid in this dissection. This eye RFP reporter is only expressed in the eye and can be visualized using fluorescence microscopy either in live tissue or after fixation without the need for signal amplification. We also show how this reporter can be used to identify specific cells types within the eye disc. This protocol and the use of the eye RFP reporter will aid researchers using the Drosophila eye to address fundamentally important biological questions. PMID- 20010544 TI - Circulating methylarginine levels and the decline in renal function in patients with chronic kidney disease are modulated by DDAH1 polymorphisms. AB - In patients with chronic kidney disease, high plasma levels of the endogenous nitric oxide synthase inhibitor, asymmetric dimethylarginine, are thought to contribute to decline in renal function. Here we took a candidate gene approach to determine any causal role of asymmetric dimethylarginine in the progression of chronic kidney disease. The impact of single-nucleotide polymorphisms in the genes encoding the two isoforms of the asymmetric dimethylarginine-degrading enzyme, dimethylarginine dimethylaminohydrolase (DDAH1 and DDAH2), on enzyme expression, plasma asymmetric dimethylarginine levels, and longitudinal changes in estimated glomerular filtration rate were determined in various patient groups. There was evidence suggesting that the rs17384213 DDAH1 GG genotype was associated with increased expression of its mRNA in kidney allografts. Healthy subjects carrying the rs17384213 G allele had lower plasma asymmetric dimethylarginine, and a similar borderline association was found in patients with chronic kidney disease. This allele, however, was independently associated with a steeper decline in renal function in two separate cohorts of patients with chronic kidney disease. We conclude that polymorphisms in DDAH1 alter the rate of decline of glomerular filtration rate in subjects with chronic kidney disease. Our findings show that DDAH1 modulates plasma asymmetric dimethylarginine and contributes to the decline in renal function. However, it appears that increases in circulating methylarginine did not mediate progressive kidney injury. PMID- 20010545 TI - Caring for adolescent renal patients. AB - Adolescent patients who require the care of a nephrologist present with a variety of challenges. For the treating physician, it is important to be competent in relevant principles of adolescent physiology, such as somatic growth and puberty. It is also essential to be familiar with the stages of adolescent emotional and intellectual development, and their influence on behavior and adherence. Patients transitioning from pediatric nephrology may experience difficulties adjusting to the adult care system. Although the renal pathology of adolescents has more in common with that of adults than that of prepubertal children, there are unique epidemiological, diagnostic, and treatment approaches to these patients that should be considered. Membranous glomerulonephritis and diabetic nephropathy are rare in adolescents, whereas IgA and systemic lupus erythematosus are often important differential diagnoses. Despite all of the challenges posed by adolescents, there is a real prospect of intervening early in the course of chronic kidney disease with the potential to alleviate long-term morbidity. PMID- 20010546 TI - FGF23-parathyroid interaction: implications in chronic kidney disease. AB - Over the past few years there have been considerable advances in our understanding of the physiological regulation of mineral homeostasis. One of the most important breakthroughs is the identification of fibroblastic growth factor 23 (FGF23) and its role as a key regulator of phosphate and 1,25-dihydroxyvitamin D metabolism. FGF23 exerts its biological functions by binding to its cognate receptor in the presence of Klotho as a cofactor. FGF23 principally acts on the kidney to induce urinary phosphate excretion and suppresses 1,25-dihydroxyvitamin D synthesis, thereby indirectly modulating parathyroid hormone secretion. FGF23 also acts directly on the parathyroid to decrease parathyroid hormone synthesis and secretion. In patients with chronic kidney disease, FGF23 levels increase progressively to compensate for phosphate retention, but these elevated FGF23 levels fail to suppress the secretion of parathyroid hormone, particularly in the setting of uremia. Recent data suggest that this parathyroid resistance to FGF23 may be caused by decreased expression of Klotho-FGFR1 complex in hyperplastic parathyroid glands. This review summarizes recent insights into the role of FGF23 in mineral homeostasis and discusses the involvement of its direct and indirect interaction with the parathyroid gland, particularly focusing on the pathophysiology of secondary hyperparathyroidism in chronic kidney disease. PMID- 20010547 TI - The MILLER banding procedure is an effective method for treating dialysis associated steal syndrome. AB - We evaluated the efficacy of the Minimally Invasive Limited Ligation Endoluminal Assisted Revision (MILLER) banding procedure in treating dialysis-associated steal syndrome or high-flow access problems. A retrospective analysis was conducted, evaluating banding of 183 patients of which 114 presented with hand ischemia (Steal) and 69 with clinical manifestations of pathologic high access flow such as congestive heart failure. Patients were assessed for technical success and symptomatic improvement, primary and secondary access patency, and primary band patency. Overall, 183 patients underwent a combined 229 bandings with technical success achieved in 225. Complete symptomatic relief (clinical success) was attained in 109 Steal patients and in all high-flow patients. The average follow-up time was 11 months with a 6-month primary band patency of 75 and 85% for Steal and high-flow patients, respectively. At 24 months the secondary access patency was 90% and the thrombotic event rates for upper-arm fistulas, forearm fistulas, and grafts were 0.21, 0.10, and 0.92 per access-year, respectively. Hence, the minimally invasive MILLER procedure appears to be an effective and durable option for treating dialysis access-related steal syndrome and high-flow-associated symptoms. PMID- 20010548 TI - Upregulation of Hic-5 in glomerulosclerosis and its regulation of mesangial cell apoptosis. AB - Glomerulosclerosis is characterized by the loss of glomerular cells by apoptosis and deposition of collagen type I into the normal collagen IV-containing mesangial matrix. We sought to determine the alterations that might contribute to these changes by performing proteomic analysis of rat mesangial cell lysates comparing cells cultured on normal collagen type IV to those grown on abnormal collagen type I surfaces. Subculture on collagen type I was associated with changed expression of several proteins, including a significant upregulation of the paxillin-like LIM protein, hydrogen-peroxide-induced clone 5 (Hic-5), and increased the susceptibility of the cells to apoptosis in response to physiological triggers. When we knocked down Hic-5 (using siRNA), we found mesangial cells grown on collagen type I were protected from apoptosis to the same degree as untreated cells grown on collagen type IV. Further we found that the level of Hic-5 in vivo was almost undetectable in control rats but increased dramatically in the glomerular mesangium of remnant kidneys 90 and 120 days after subtotal nephrectomy. This induction of Hic-5 paralleled the upregulation of mesangial collagen type I expression and glomerular cell apoptosis. Our results suggest that Hic-5 is pivotal in mediating the response of mesangial cells to attachment on abnormal extracellular matrix during glomerular scarring. PMID- 20010549 TI - Serotonin depletion hampers survival and proliferation in neurospheres derived from adult neural stem cells. AB - Serotonin (5-HT) and the serotonergic system have recently been indicated as modulators of adult hippocampal neurogenesis. In this study, we evaluated the role of 5-HT on the functional features in neurospheres derived from adult neural stem cells (ANSC). We cultured neurospheres derived from mouse hippocampus in serum-free medium containing epidermal (EGF) and type-2 fibroblast growth factor (FGF2). Under these conditions ANSC expressed both isoforms of tryptophane hydroxylase (TPH) and produced 5-HT. Blocking TPH function by para chlorophenylalanine (PCPA) reduced ANSC proliferation, which was rescued by exogenous 5-HT. 5-HT action on ANSC was mediated predominantly by the serotonin receptor subtype 5-HT1A and, to a lesser extent, through the 5-HT2C (receptor) subtype, as shown by selectively antagonizing these receptors. Finally, we documented a 5-HT-induced increase of ANSC migration activity. In summary, we demonstrated a powerful serotonergic impact on ANSC functional features, which was mainly mediated by 5-HT1A receptors. PMID- 20010550 TI - Chronic cocaine-induced H3 acetylation and transcriptional activation of CaMKIIalpha in the nucleus accumbens is critical for motivation for drug reinforcement. AB - The regulation of gene expression in the brain reward regions is known to contribute to the pathogenesis and persistence of drug addiction. Increasing evidence suggests that the regulation of gene transcription is mediated by epigenetic mechanisms that alter the chromatin structure at specific gene promoters. To better understand the involvement of epigenetic regulation in drug reinforcement properties, rats were subjected to cocaine self-administration paradigm. Daily histone deacetylase (HDAC) inhibitor infusions in the shell of the nucleus accumbens (NAc) caused an upward shift in the dose-response curve under fixed-ratio schedule and increased the break point under progressive-ratio schedule, indicating enhanced motivation for self-administered drug. The effect of the HDAC inhibitor is attributed to the increased elevation of histone acetylation induced by chronic, but not acute, cocaine experience. In contrast, neutralizing the chronic cocaine-induced increase in histone modification by the bilateral overexpression of HDAC4 in the NAc shell reduced drug motivation. The association between the motivation for cocaine and the transcriptional activation of addiction-related genes by H3 acetylation in the NAc shell was analyzed. Among the genes activated by chronic cocaine experiences, the expression of CaMKIIalpha, but not CaMKIIbeta, correlated positively with motivation for the drug. Lentivirus-mediated shRNA knockdown experiments showed that CaMKIIalpha, but not CaMKIIbeta, in the NAc shell is essential for the maintenance of motivation to self-administered cocaine. These findings suggest that chronic drug use-induced transcriptional activation of genes, such as CaMKIIalpha, modulated by H3 acetylation in the NAc is a critical regulatory mechanism underlying motivation for drug reinforcement. PMID- 20010551 TI - Divalproex sodium vs placebo for the treatment of irritability in children and adolescents with autism spectrum disorders. AB - Autism spectrum disorders (ASDs) are neurodevelopmental disorders characterized by social and language deficits and by repetitive behaviors and interests. Irritability/aggression is a significant comorbid symptom in this population, which greatly impacts burden of care. This study examined the effect of divalproex sodium for irritability/aggression in children and adolescents with ASD. This was a 12-week randomized, double-blind, placebo-controlled trial. All efficacy measures were obtained by an independent evaluator blinded to randomization condition and side effects. A total of 55 subjects gavetheir consent and 27 were randomized in a 1 : 1 manner (mean age 9.46+/-2.46, mean nonverbal IQ 63.3+/-23.9). Two subjects from the active group and one subject from the placebo group discontinued the study because of either a lack of efficacy or side effects (increased irritability). Primary outcome measures were Aberrant Behavior Checklist-Irritability subscale and Clinical Global Impression Improvement, which focused on irritability. Overall, 62.5% of divalproex subjects vs 9% of placebo subjects were responders (CGI-irritability OR: 16.7, Fisher's exact p=0.008). A statistically significant improvement was also noted on the ABC Irritability subscale (p=0.048). There was a trend for responders to have higher valproate blood levels compared with nonresponders. This study suggests the efficacy of divalproex for the treatment of irritability in children and adolescents with ASD. Larger sample follow-up studies are warranted. PMID- 20010552 TI - Individual and additive effects of the CNR1 and FAAH genes on brain response to marijuana cues. AB - As previous work has highlighted the significance of the cannabinoid receptor 1 (CNR1) and fatty acid amide hydrolase (FAAH) genes with respect to cannabis dependence (CD), this study sought to characterize the neural mechanisms that underlie these genetic effects. To this end, we collected DNA samples and fMRI data using a cue-elicited craving paradigm in thirty-seven 3-day-abstinent regular marijuana users. The participants were grouped according to their genotype on two single-nucleotide polymorphisms (SNPs) earlier associated with CD phenotypes: rs2023239 in CNR1 and rs324420 in FAAH. Between-group comparisons showed that carriers of the CNR1 rs2023239 G allele had significantly greater activity in reward-related areas of the brain, such as the orbitofrontal cortex (OFC), inferior frontal gyrus (IFG), and anterior cingulate gyrus (ACG), during exposure to marijuana cues, as compared with those with the A/A genotype for this SNP. The FAAH group contrasts showed that FAAH rs324420 C homozygotes also had greater activation in widespread areas within the reward circuit, specifically in the OFC, ACG, and nucleus accumbens (NAc), as compared with the FAAH A-allele carriers. Moreover, there was a positive correlation between neural response in OFC and NAc and the total number of risk alleles (cluster-corrected p<0.05). These findings are in accord with earlier reported associations between CNR1 and FAAH and CD intermediate phenotypes, and suggest that the underlying mechanism of these genetic effects may be enhanced neural response in reward areas of the brain in carriers of the CNR1 G allele and FAAH C/C genotype in response to marijuana cues. PMID- 20010554 TI - The gap between clinical trials and clinical practice: the use of pragmatic clinical trials to inform regulatory decision making. PMID- 20010553 TI - Inhibitors of class 1 histone deacetylases reverse contextual memory deficits in a mouse model of Alzheimer's disease. AB - Alzheimer's disease (AD) is a neurodegenerative disorder characterized clinically by cognitive impairments that progress to dementia and death. The earliest symptoms of AD present as a relatively pure deficit in memory retrieval. Therefore, drug treatments that intervene in the early stages of AD by rescuing memory deficits could be promising therapies to slow, or even reverse progression of the disease. In this study, we tested the potential of systemic histone deacetylase inhibitor (HDACi) treatment to rescue cognitive deficits in a mouse model of AD. APPswe/PS1dE9 mice showed pronounced contextual memory impairments beginning at 6 months of age. Chronic HDACi injections (2-3 weeks) did not alter contextual memory formation in normal mice, but had profound effects in transgenic animals. Injections of sodium valproate, sodium butyrate, or vorinostat (suberoylanilide hydroxamic acid; Zolinza) completely restored contextual memory in these mutant mice. Further behavioral testing of the HDACi treated transgenic mice showed that the newly consolidated memories were stably maintained over a 2-week period. Measurement of the HDAC isoform selectivity profile of sodium valproate, sodium butyrate, and vorinostat revealed the common inhibition of class I HDACs (HDAC1, 2, 3, 8) with little effect on the class IIa HDAC family members (HDAC4, 5, 7, 9) and inhibition of HDAC6 only by vorinostat. These preclinical results indicate that targeted inhibition of class I HDAC isoforms is a promising avenue for treating the cognitive deficits associated with early stage AD. PMID- 20010557 TI - Unbottling the genes. PMID- 20010555 TI - A dosing/cross-development study of the multikinase inhibitor sorafenib in patients with pulmonary arterial hypertension. AB - Pulmonary arterial hypertension (PAH) and cancer share elements of pathophysiology. This provides an opportunity for the cross-development of anticancer agents that can be used in improving PAH care. The adaptation of new drugs across these disease populations warrants a structured approach. This study was a 16-week, phase Ib, single-center, open-label trial of the multikinase/angiogenesis inhibitor sorafenib. In order to assess the safety of sorafenib in PAH, patients with advanced but stable disease on parenteral prostanoids (with or without oral sildenafil) were initiated on treatment at the lowest active dosage administered to cancer patients: 200 mg daily. Patients underwent weekly clinical evaluations and monthly functional testing and dose escalations to a final dosage of 400 mg twice daily. Among 12 patients (10 of them women), sorafenib was well tolerated at 200 mg twice daily. The most common adverse events were moderate skin reactions on the hands and feet and alopecia. Our conclusion was therefore that this is a tolerable dosing regimen for testing the therapeutic activity of sorafenib in PAH patients. PMID- 20010556 TI - Evaluation and management of cardiac safety using the electrocardiogram in oncology clinical trials: focus on cardiac repolarization (QTc interval). AB - Non-antiarrhythmic drugs have been reported to prolong the QTc interval and induce potentially fatal ventricular tachyarrhythmias. An increasing number of drugs that are used for treating malignancies are no exception. Therefore, both oncologists and regulators expect sponsors of oncology drugs to evaluate, during the development of the drugs, their effects on the electrocardiogram (ECG), particularly on the QTc interval. In the case of agents that cannot be administered to healthy volunteers, the primary approach is to carry out an intense ECG evaluation, employing robust ECG recordings, during early-phase clinical trials, together with characterization of the concentration-QTc interval relationship, and follow this up with an appropriate intensity of ECG monitoring in the later phases of development. This article describes the broad principles of these approaches, including recommendations for exclusion criteria (relative to baseline QTc interval and to cardiac comorbidity); it also describes methods for conducting ECG monitoring and a proposed scheme for the management of any QTc related effects that may emerge. PMID- 20010558 TI - Federal stimulus adds momentum to sequencing stampede. PMID- 20010559 TI - ARRA boosts cancer programs but impact on personalized medicine unclear. PMID- 20010560 TI - Brazil and India access pharma vaccine know-how. PMID- 20010561 TI - Gates pours cash into agriculture. PMID- 20010562 TI - Congress punts on SBIR reauthorization. PMID- 20010563 TI - Biotechs get $1 billion windfall in R&D tax credits. PMID- 20010566 TI - FDA recruits prominent critics. PMID- 20010567 TI - Comparative effectiveness $100 million handout. PMID- 20010568 TI - REMS violations fines. PMID- 20010572 TI - Proteostasis therapeutics. PMID- 20010573 TI - What's in a name? PMID- 20010575 TI - Biotech in the basement. PMID- 20010574 TI - Making green. PMID- 20010576 TI - Medicare formulary coverage for top-selling biologics. PMID- 20010577 TI - Biotech and gender issues in the developing world. PMID- 20010578 TI - The facts about Stemedica. PMID- 20010580 TI - First transgenic geminivirus-resistant plant in the field. PMID- 20010582 TI - The changing economics of DNA synthesis. PMID- 20010583 TI - Parts, property and sharing. PMID- 20010584 TI - Building outside of the box: iGEM and the BioBricks Foundation. PMID- 20010585 TI - Our synthetic future. PMID- 20010586 TI - Should moral objections to synthetic biology affect public policy? PMID- 20010587 TI - From synthetic biology to biohacking: are we prepared? PMID- 20010588 TI - Engineering a new business. PMID- 20010589 TI - The sorcerer of synthetic genomes. PMID- 20010590 TI - The obviousness rejection as a barrier to biotech patent prosecution. PMID- 20010592 TI - Engineering direct conversion of CO(2) to biofuel. PMID- 20010593 TI - A first look at entire human methylomes. PMID- 20010594 TI - Capturing global metabolism. PMID- 20010596 TI - How does multiple testing correction work? PMID- 20010598 TI - Genome engineering. AB - For more than 50 years, those engineering genetic material have pursued increasingly challenging targets. During that time, the tools and resources available to the genetic engineer have grown to encompass new extremes of both scale and precision, opening up new opportunities in genome engineering. Today, our capacity to generate larger de novo assemblies of DNA is increasing at a rapid pace (with concomitant decreases in manufacturing cost). We are also witnessing potent demonstrations of the power of merging randomness and selection with engineering approaches targeting large numbers of specific sites within genomes. These developments promise genetic engineering with unprecedented levels of design originality and offer new avenues to expand both our understanding of the biological world and the diversity of applications for societal benefit. PMID- 20010597 TI - Next-generation synthetic gene networks. AB - Synthetic biology is focused on the rational construction of biological systems based on engineering principles. During the field's first decade of development, significant progress has been made in designing biological parts and assembling them into genetic circuits to achieve basic functionalities. These circuits have been used to construct proof-of-principle systems with promising results in industrial and medical applications. However, advances in synthetic biology have been limited by a lack of interoperable parts, techniques for dynamically probing biological systems and frameworks for the reliable construction and operation of complex, higher-order networks. As these challenges are addressed, synthetic biologists will be able to construct useful next-generation synthetic gene networks with real-world applications in medicine, biotechnology, bioremediation and bioenergy. PMID- 20010603 TI - Stomagen positively regulates stomatal density in Arabidopsis. AB - Stomata in the epidermal tissues of leaves are valves through which passes CO(2), and as such they influence the global carbon cycle. The two-dimensional pattern and density of stomata in the leaf epidermis are genetically and environmentally regulated to optimize gas exchange. Two putative intercellular signalling factors, EPF1 and EPF2, function as negative regulators of stomatal development in Arabidopsis, possibly by interacting with the receptor-like protein TMM. One or more positive intercellular signalling factors are assumed to be involved in stomatal development, but their identities are unknown. Here we show that a novel secretory peptide, which we designate as stomagen, is a positive intercellular signalling factor that is conserved among vascular plants. Stomagen is a 45-amino rich peptide that is generated from a 102-amino-acid precursor protein designated as STOMAGEN. Both an in planta analysis and a semi-in-vitro analysis with recombinant and chemically synthesized stomagen peptides showed that stomagen has stomata-inducing activity in a dose-dependent manner. A genetic analysis showed that TMM is epistatic to STOMAGEN (At4g12970), suggesting that stomatal development is finely regulated by competitive binding of positive and negative regulators to the same receptor. Notably, STOMAGEN is expressed in inner tissues (the mesophyll) of immature leaves but not in the epidermal tissues where stomata develop. This study provides evidence of a mesophyll-derived positive regulator of stomatal density. Our findings provide a conceptual advancement in understanding stomatal development: inner photosynthetic tissues optimize their function by regulating stomatal density in the epidermis for efficient uptake of CO(2). PMID- 20010599 TI - Synthetic viruses: a new opportunity to understand and prevent viral disease. AB - Rapid progress in DNA synthesis and sequencing is spearheading the deliberate, large-scale genetic alteration of organisms. These new advances in DNA manipulation have been extended to the level of whole-genome synthesis, as evident from the synthesis of poliovirus, from the resurrection of the extinct 1918 strain of influenza virus and of human endogenous retroviruses and from the restructuring of the phage T7 genome. The largest DNA synthesized so far is the 582,970 base pair genome of Mycoplasma genitalium, although, as yet, this synthetic DNA has not been 'booted' to life. As genome synthesis is independent of a natural template, it allows modification of the structure and function of a virus's genetic information to an extent that was hitherto impossible. The common goal of this new strategy is to further our understanding of an organism's properties, particularly its pathogenic armory if it causes disease in humans, and to make use of this new information to protect from, or treat, human viral disease. Although only a few applications of virus synthesis have been described as yet, key recent findings have been the resurrection of the 1918 influenza virus and the generation of codon- and codon pair-deoptimized polioviruses. PMID- 20010604 TI - Preparation and detection of a mechanical resonator near the ground state of motion. AB - Cold, macroscopic mechanical systems are expected to behave contrary to our usual classical understanding of reality; the most striking and counterintuitive predictions involve the existence of states in which the mechanical system is located in two places simultaneously. Various schemes have been proposed to generate and detect such states, and all require starting from mechanical states that are close to the lowest energy eigenstate, the mechanical ground state. Here we report the cooling of the motion of a radio-frequency nanomechanical resonator by parametric coupling to a driven, microwave-frequency superconducting resonator. Starting from a thermal occupation of 480 quanta, we have observed occupation factors as low as 3.8 +/- 1.3 and expect the mechanical resonator to be found with probability 0.21 in the quantum ground state of motion. Further cooling is limited by random excitation of the microwave resonator and heating of the dissipative mechanical bath. This level of cooling is expected to make possible a series of fundamental quantum mechanical observations including direct measurement of the Heisenberg uncertainty principle and quantum entanglement with qubits. PMID- 20010605 TI - Time-reversal symmetry breaking and spontaneous Hall effect without magnetic dipole order. AB - Spin liquids are magnetically frustrated systems, in which spins are prevented from ordering or freezing, owing to quantum or thermal fluctuations among degenerate states induced by the frustration. Chiral spin liquids are a hypothetical class of spin liquids in which the time-reversal symmetry is macroscopically broken in the absence of an applied magnetic field or any magnetic dipole long-range order. Even though such chiral spin-liquid states were proposed more than two decades ago, an experimental realization and observation of such states has remained a challenge. One of the characteristic order parameters in such systems is a macroscopic average of the scalar spin chirality, a solid angle subtended by three nearby spins. In previous experimental reports, however, the spin chirality was only parasitic to the non-coplanar spin structure associated with a magnetic dipole long-range order or induced by the applied magnetic field, and thus the chiral spin-liquid state has never been found. Here, we report empirical evidence that the time-reversal symmetry can be broken spontaneously on a macroscopic scale in the absence of magnetic dipole long-range order. In particular, we employ the anomalous Hall effect to directly probe the broken time-reversal symmetry for the metallic frustrated magnet Pr(2)Ir(2)O(7). An onset of the Hall effect is observed at zero field in the absence of uniform magnetization, within the experimental accuracy, suggesting an emergence of a chiral spin liquid. The origin of this spontaneous Hall effect is ascribed to chiral spin textures, which are inferred from the magnetic measurements indicating the spin ice-rule formation. PMID- 20010606 TI - Preventing the return of fear in humans using reconsolidation update mechanisms. AB - Recent research on changing fears has examined targeting reconsolidation. During reconsolidation, stored information is rendered labile after being retrieved. Pharmacological manipulations at this stage result in an inability to retrieve the memories at later times, suggesting that they are erased or persistently inhibited. Unfortunately, the use of these pharmacological manipulations in humans can be problematic. Here we introduce a non-invasive technique to target the reconsolidation of fear memories in humans. We provide evidence that old fear memories can be updated with non-fearful information provided during the reconsolidation window. As a consequence, fear responses are no longer expressed, an effect that lasted at least a year and was selective only to reactivated memories without affecting others. These findings demonstrate the adaptive role of reconsolidation as a window of opportunity to rewrite emotional memories, and suggest a non-invasive technique that can be used safely in humans to prevent the return of fear. PMID- 20010607 TI - Phylogenies reveal new interpretation of speciation and the Red Queen. AB - The Red Queen describes a view of nature in which species continually evolve but do not become better adapted. It is one of the more distinctive metaphors of evolutionary biology, but no test of its claim that speciation occurs at a constant rate has ever been made against competing models that can predict virtually identical outcomes, nor has any mechanism been proposed that could cause the constant-rate phenomenon. Here we use 101 phylogenies of animal, plant and fungal taxa to test the constant-rate claim against four competing models. Phylogenetic branch lengths record the amount of time or evolutionary change between successive events of speciation. The models predict the distribution of these lengths by specifying how factors combine to bring about speciation, or by describing how rates of speciation vary throughout a tree. We find that the hypotheses that speciation follows the accumulation of many small events that act either multiplicatively or additively found support in 8% and none of the trees, respectively. A further 8% of trees hinted that the probability of speciation changes according to the amount of divergence from the ancestral species, and 6% suggested speciation rates vary among taxa. By comparison, 78% of the trees fit the simplest model in which new species emerge from single events, each rare but individually sufficient to cause speciation. This model predicts a constant rate of speciation, and provides a new interpretation of the Red Queen: the metaphor of species losing a race against a deteriorating environment is replaced by a view linking speciation to rare stochastic events that cause reproductive isolation. Attempts to understand species-radiations or why some groups have more or fewer species should look to the size of the catalogue of potential causes of speciation shared by a group of closely related organisms rather than to how those causes combine. PMID- 20010611 TI - Assessment of the acute effects of tadalafil on the cardiovascular system based on examination of serum oxidative status and paraoxonase activity in men with erectile dysfunction: a preliminary study. AB - Phosphodiesterase type 5 inhibitors were initially approved for the treatment of erectile dysfunction in men and were later suggested for some systemic disorders, mostly due to their possible beneficial effects on endothelial functions. Paradoxically, though, phosphodiesterase type 5 inhibitors may have some life threatening effects, for which there is weak evidence, it appears that they are associated with cardiovascular problems such as myocardial infarction and stroke. This study aimed to investigate the acute effects of tadalafil citrate on the cardiovascular system by evaluating serum oxidative status and paraoxonase-1 activity. Sera of 36 patients with erectile dysfunction were analyzed for total antioxidant status, total oxidant status and paraoxonase-1, before and after the administration of tadalafil citrate. Pre- and post-tadalafil citrate serum levels of total antioxidants, total oxidants and paraoxonase-1 were 1.1+/-0.0 and 1.6+/ 0.0 micromol H(2)O(2) equiv l(-1), 10.3+/-1.1 and 6.9+/-1.2 micromol H(2)O(2) equiv l(-1), and 111.6+/-17.8 and 168.0+/-18.1 U l(-1), respectively (P<0.0001 for all results). This preliminary study confirmed that tadalafil citrate exerts a beneficial acute effect on the cardiovascular system by reducing serum levels of oxidative stress and increasing serum levels of paraoxonase-1. PMID- 20010608 TI - Evaluation of cadmium, lead, nickel and zinc status in biological samples of smokers and nonsmokers hypertensive patients. AB - The objective of this study was to evaluate the association between trace and toxic elements zinc (Zn), cadmium (Cd), nickel (Ni) and lead (Pb) in biological samples (scalp hair, blood and urine) of smoker and nonsmoker hypertensive patients (n=457), residents of Hyderabad, Pakistan. For the purpose of comparison, the biological samples of age-matched healthy controls were selected as referents. The concentrations of trace and toxic elements were measured by atomic absorption spectrophotometer prior to microwave-assisted acid digestion. The validity and accuracy of the methodology were checked using certified reference materials and by the conventional wet acid digestion method on the same certified reference materials and real samples. The recovery of all the studied elements was found to be in the range of 97.8-99.3% in certified reference materials. The results of this study showed that the mean values of Cd, Ni and Pb were significantly higher in scalp hair, blood and urine samples of both smoker and nonsmoker patients than in referents (P<0.001), whereas the concentration of Zn was lower in the scalp hair and blood, but higher in the urine samples of hypertensive patients. The deficiency of Zn and the high exposure of toxic metals as a result of tobacco smoking may be synergistic with risk factors associated with hypertension. PMID- 20010612 TI - The impact of a noise reduction quality improvement project upon sound levels in the open-unit-design neonatal intensive care unit. AB - OBJECTIVE: To decrease measured sound levels in the neonatal intensive care unit through implementation of human factor and minor design modification strategies. STUDY DESIGN: Prospective time series. Two open-unit-design neonatal centers. RESULT: Implementation of a coordinated program of noise reduction strategies did not result in any measurable improvement in levels of loudness or quiet. CONCLUSION: Two centers, using primarily human behavior noise reduction strategies, were unable to demonstrate measurable improvements in sound levels within the occupied open-unit-design neonatal intensive care unit. PMID- 20010613 TI - A silver-alginate-coated dressing to reduce peripherally inserted central catheter (PICC) infections in NICU patients: a pilot randomized controlled trial. AB - OBJECTIVE: Our aim was to evaluate the safety of a silver-alginate-containing dressing to reduce peripherally inserted central catheter (PICC) infections in neonatal intensive care unit (NICU) patients. STUDY DESIGN: Patients were randomized 3:1 to receive a patch containing silver, alginate and maltodextrin or standard of care. Patches were placed under the regular transparent retention dressing at the PICC exit site at insertion and were replaced with every dressing change at least every 2 weeks until PICC discontinuation. All study infants were monitored for adverse skin reactions. RESULT: A total of 100 infants were followed up for 1922 person-days, including 75 subjects with 89 PICCs who received the patch. The median birth weight (1330 g) and median gestational age (30 weeks) was lower in the patch group when compared with the controls (P=0.001 and 0.005, respectively). Study patients received the patch with their PICC at a median age of 5 days; the patch stayed in place for a median of 13 days. We noted no adverse skin reactions and found no evidence that the patch alters the microbiology of PICC-associated infections. CONCLUSION: This pilot trial suggests that silver-alginate-coated dressings are skin safe and their inclusion in future trials aimed at reduction of PICC-associated bloodstream infections in the NICU should be considered. PMID- 20010614 TI - Nurses' compliance with alarm limits for pulse oximetry: qualitative study. AB - OBJECTIVE: Monthly audits for the multicenter Canadian Oxygen Trial have shown that our neonatal team has consistently maintained study participants within the intended pulse oximetry alarm limits between 68 and 79% of the time during the first 3 days of life while infants were receiving supplemental oxygen. This good performance prompted us to explore our nurses' perceptions of what makes them compliant with alarm limits for pulse oximetry in preterm infants. STUDY DESIGN: The local neonatal research nurse interviewed all 41 staff nurses in the neonatal intensive care unit at the University Children's Hospital in Tuebingen, Germany, for this qualitative study. RESULT: Nurses identified education (22/41), prompt response to alarms (22/41), a favorable patient-to-staff ratio (18/41), root cause analysis at the bedside (15/41), and the high priority given to control of oxygen therapy in the department (14/41) as the five most important determinants of their good compliance. CONCLUSION: These findings may be useful for other neonatal teams who struggle to improve their compliance with alarm limits for pulse oximetry. PMID- 20010615 TI - Effects of instrumental dead space reduction during weaning from synchronized ventilation in preterm infants. AB - OBJECTIVE: A majority of the modalities of synchronized ventilation in preterm infants require the use of flow sensors that can increase dead space and may adversely affect ventilator weaning. The objective of this study was to assess the effects of flow sensor dead space during synchronized intermittent mandatory ventilation (SIMV) weaning in preterm infants. STUDY DESIGN: Twelve preterm infants (gestational age 25+/-2 weeks, birth weight 705+/-158 g, age: 31+/-186 days, SIMV rate: 25+/-8 breaths min(-1), peak inspiratory pressure 18+/-2 cm H(2)O, positive end-expiratory pressure: 5+/-0.5 cm H(2)O, pressure support: 9+/ 3 cm H(2)O, fraction of inspired oxygen: 34+/-6%) underwent two 2.5-h weaning periods during which SIMV rate was reduced twice by 5 breaths min(-1) at 30-min intervals as tolerated, with and without reduction of flow sensor dead space, in random sequence. A 30-min baseline was obtained before each weaning period. Dead space was reduced by flushing the flow sensor with a continuous gas leak flow in the endotracheal tube connector. RESULT: Transcutaneous CO(2) tension during SIMV weaning periods without and with reduced dead space did not differ from baseline, whereas total minute ventilation and tidal volume were lower during the SIMV weaning period with reduced dead space. Three infants did not tolerate SIMV weaning without while one infant did not tolerate weaning with reduced dead space. CONCLUSION: SIMV weaning elicited a compensatory rise in spontaneous ventilation. When flow sensor dead space was reduced during SIMV weaning, gas exchange was maintained with lower minute ventilation. Instrumental dead space imposes a ventilatory burden during SIMV weaning in small preterm infants. PMID- 20010616 TI - Early cortisol values and long-term outcomes in extremely low birth weight infants. AB - OBJECTIVE: Both excess and insufficient levels of glucocorticoid in extremely low birth weight (ELBW) infants have been associated with adverse hospital outcomes, whereas excess glucocorticoid exposure has been associated with long-term adverse neurodevelopment. Our objective was to evaluate the relationship between neonatal cortisol concentrations and long-term outcomes of growth and neurodevelopment. STUDY DESIGN: As part of a multicenter randomized trial of hydrocortisone treatment for prophylaxis of relative adrenal insufficiency, cortisol concentrations were obtained at 12 to 48 h of postnatal age and at days 5 to 7 on 350 intubated ELBW infants, of whom 252 survived and returned for neurodevelopmental follow-up at 18 to 22 months corrected age. Cortisol values from each time point were divided into quartiles. Growth and neurodevelopmental outcome were compared for each quartile. RESULT: Median cortisol value was 16.0 microg per 100 ml at baseline for all infants, and 13.1 microg per 100 ml on days 5 to 7 in the placebo group. Outcomes did not differ in each quartile between treatment and placebo groups. Low cortisol values at baseline or at days 5 to 7 were not associated with impaired growth or neurodevelopment at 18 to 22 months corrected age. High cortisol values were associated with an increase in cerebral palsy, related to the increased incidence of severe intraventricular hemorrhage (IVH) and periventricular leukomalacia. CONCLUSION: Low cortisol concentrations were not predictive of adverse long-term outcomes. High cortisol concentrations, although predictive of short-term adverse outcomes such as IVH and periventricular leukomalacia, did not additionally predict adverse outcome. Further analysis into identifying factors that modulate cortisol concentrations shortly after birth could improve our ability to identify those infants who are most likely to benefit from treatment with hydrocortisone. PMID- 20010617 TI - Plasma adiponectin concentration is associated with ambulatory daytime systolic blood pressure but not with the dipping status. AB - The objective of this study was to analyse the relationship between the ambulatory blood pressure (ABP) measurement and plasma adiponectin levels in a population-based cohort. Non-hypertensive, non-diabetics from the Oulu Project Elucidating Risk of Atherosclerosis cohort aged 40-60 years with ABP measurement available in 226 men and 236 women were analysed. ABP was recorded using the fully automatic SpaceLabs 90207 oscillometric unit. Plasma adiponectin concentrations were assayed using the enzyme-linked immunosorbent assay method. Without adjustment the highest plasma adiponectin tertile was associated with the lowest ABP and office BP measurements (P from 0.025 to P<0.001, respectively). Only the association of plasma adiponectin concentration with systolic ABP was independent of other conventional risk factors (age, body mass index (BMI), waist, gender, insulin sensitivity index, smoking and alcohol consumption) for hypertension (P=0.017). No association was observed between systolic dipping pattern and adiponectin level. The plasma high adiponectin concentration is independently associated with low daytime systolic ABP value. The mechanisms may include effects on endothelial function and the sympathetic nervous system. PMID- 20010618 TI - Effectiveness of initiating treatment with valsartan/hydrochlorothiazide in patients with stage-1 or stage-2 hypertension. AB - This prospective, 6-week, multicenter, double-blind study examined the benefits of initiating treatment with combination valsartan/hydrochlorothiazide (HCTZ) compared with initial valsartan monotherapy for 648 patients with stage-1 or stage-2 hypertension (age=52.6+/-10 years; 54% male; baseline blood pressure (BP)=161/98 mm Hg, 32% stage 1). Patients were randomized to valsartan 80 mg (V low), valsartan 160 mg (V-high) or valsartan/HCTZ 160/12.5 mg (V/HCTZ), and electively titrated after weeks 2 and 4 to the next dosage level (maximum dose valsartan/HCTZ 160/25 mg) if BP remained >140/90 mm Hg. At end of the study, patients initiated with V/HCTZ required less titration steps compared with the initial valsartan monotherapy groups (63 vs 86% required titration by study end, respectively) and reached the target BP goal of <140/90 mm Hg in a shorter period of time (2.8 weeks) (P<0.0001) vs V-low (4.3 weeks) and V-high (3.9 weeks). Initial combination therapy was also associated with higher BP control rates and greater reductions in both systolic and diastolic BP from baseline (63%, -27.7+/ 13/-15.1+/-8 mm Hg) compared with V-low (46%, -21.2+/-13/-11.4+/-8 mm Hg, P<0.0001) or V-high (51%, -24.0+/-13/-12.0+/-10 mm Hg, P<0.01). Overall and drug related AEs were mild to moderate and were similar between V/HCTZ (53.1 and 14.1%, respectively) and the two monotherapy groups, V-low (50.5 and 13.8%) and V high (50.7 and 11.8%). In conclusion, initiating therapy with a combination of valsartan and low-dose HCTZ results in early, improved BP efficacy with similar tolerability as compared with starting treatment with a low or higher dose of valsartan for patients with stage-1 and stage-2 hypertension. PMID- 20010619 TI - Blood pressure development and hypertensive retinopathy: 20-year follow-up of middle-aged normotensive and hypertensive men. AB - Screening for hypertensive organ damage is important in assessing cardiovascular risk in hypertensive individuals. In a 20-year follow-up of normotensive and hypertensive men, signs of end-organ damage were examined, focusing on hypertensive retinopathy. In all, 56 of the original 79 men were reexamined for hypertensive organ damage, including by digital fundus photography. The diameters of the central retinal artery equivalent (CRAE) and vein were estimated and the artery-to-vein diameter ratio calculated. Components of metabolic syndrome were assessed. Fifty percent of the normotensive men developed hypertension during follow-up. Significant differences appeared in CRAE between the different blood pressure groups (P=0.025) while no differences were observed for other markers of hypertensive organ damage. There were significant relationships between CRAE and blood pressure at baseline (r=-0.466, P=0.001) and at follow-up (r=-0.508, P<0.001). A linear decrease in CRAE was observed with increasing number of components of the metabolic syndrome (beta=-3.947, R(2)=0.105, P=0.023). Retinal vascular diameters were closely linked to blood pressures and risk factors of the metabolic syndrome. The diversity in the development of hypertensive organ damage, with changes in retinal microvasculature preceding other signs of damage, should encourage more liberal use of fundus photography in assessing cardiovascular risk in hypertensive individuals. PMID- 20010621 TI - Long-term follow-up of the United Kingdom medical research council protocols for childhood acute lymphoblastic leukaemia, 1980-2001. AB - Between 1980 and 2001, the United Kingdom Medical Research Council Childhood Leukemia Working Party conducted four clinical trials in acute lymphoblastic leukaemia (ALL), which recruited a total of 6516 patients. UKALL VIII examined the role of daunorubicin in induction chemotherapy, and UKALL X examined the role of post-induction intensification. Both resulted in major improvement in the outcomes. UKALL XI examined the efficacy of different methods of central nervous system-directed therapy and the effects of an additional intensification. ALL97, which was initially based on the UKALL XD template (two intensification phases), examined the role of different steroids in induction and of different thiopurines through continuing chemotherapy. A reappraisal of results from UKALL XI compared with other cooperative group results led to a redesign in 1999, which subsequently resulted in a major improvement in outcomes. In addition, ALL97 and ALL97/99 showed a significant advantage for the use of dexamethasone rather than prednisolone; although the use of 6-thioguanine resulted in fewer relapses, this advantage was offset by an increased incidence of deaths in remission. Over the era encompassed by these four trials, there has been a major improvement in both event-free and overall survival for children in the United Kingdom with ALL. PMID- 20010622 TI - Long-term results of NOPHO ALL-92 and ALL-2000 studies of childhood acute lymphoblastic leukemia. AB - Analysis of 2668 children with acute lymphoblastic leukemia (ALL) treated in two successive Nordic clinical trials (Nordic Society of Paediatric Haematology and Oncology (NOPHO) ALL-92 and ALL-2000) showed that 75% of all patients are cured by first-line therapy, and 83% are long-term survivors. Improvements in systemic and intrathecal chemotherapy have reduced the use of central nervous system (CNS) irradiation to <10% of the patients and provided a 5-year risk of isolated CNS relapse of 2.6%. Improved risk stratification and chemotherapy have eliminated the previous independent prognostic significance of gender, CNS leukemia and translocation t(1;19)(q23;p13), whereas the post-induction level of minimal residual disease (MRD) has emerged as a new risk grouping feature. Infant leukemia, high leukocyte count, T-lineage immunophenotype, translocation t(4;11)(q21;q23) and hypodiploidy persist to be associated with lower cure rates. To reduce the overall toxicity of the treatment, including the risk of therapy related second malignant neoplasms, the current NOPHO ALL-2008 protocol does not include CNS irradiation in first remission, the dose of 6-mercaptopurine is reduced for patients with low thiopurine methyltransferase activity, and the protocol restricts the use of hematopoietic stem cell transplantation in first remission to patients without morphological remission after induction therapy or with high levels of MRD after 3 months of therapy. PMID- 20010620 TI - Long-term results of St Jude Total Therapy Studies 11, 12, 13A, 13B, and 14 for childhood acute lymphoblastic leukemia. AB - We analyzed the long-term outcome of 1011 patients treated in five successive clinical trials (Total Therapy Studies 11, 12, 13A, 13B, and 14) between 1984 and 1999. The event-free survival improved significantly (P=0.003) from the first two trials conducted in the 1980s to the three more recent trials conducted in the 1990s. Approximately 75% of patients treated in the 1980s and 80% in the 1990s were cured. Early intensive triple intrathecal therapy, together with more effective systemic therapy, including consolidation and reinduction treatment (Studies 13A and 13B) as well as dexamethasone (Study 13B), resulted in a very low rate of isolated central nervous system (CNS) relapse rate (<2%), despite the reduced use of cranial irradiation. Factors consistently associated with treatment outcome were age, leukocyte count, immunophenotype, DNA index, and minimal residual disease level after remission induction treatment. Owing to concerns about therapy-related secondary myeloid leukemia and brain tumors, in our current trials we reserve the use of etoposide for patients with refractory or relapsed leukemia undergoing hematopoietic stem cell transplantation, and cranial irradiation for those with CNS relapse. The next main challenge is to further increase cure rates while improving quality of life for all patients. PMID- 20010623 TI - Nilotinib-mediated inhibition of ABCB1 increases intracellular concentration of dasatinib in CML cells: implications for combination TKI therapy. PMID- 20010624 TI - Pirin downregulation is a feature of AML and leads to impairment of terminal myeloid differentiation. AB - Terminal differentiation of blood cells requires the concerted action of a series of transcription factors that are expressed at specific stages of maturation and function in a cell-type and dosage-dependent manner. Leukemogenic oncoproteins block differentiation by subverting the normal transcriptional status of hematopoietic precursor cells. Pirin (PIR) is a putative transcriptional regulator whose expression is silenced in cells bearing the acute myeloid leukemia-1 eight-twenty-one (AML1/ETO) and promyelocytic leukemia/retinoic acid receptor (PML/RAR) leukemogenic fusion proteins. A role for PIR in myeloid differentiation has not to date been reported. In this study we show that PIR expression is significantly repressed in a large proportion of acute myeloid leukemias (AMLs), regardless of subtype or underlying karyotypic abnormalities. We show that PIR expression increases during in vitro myeloid differentiation of primary hematopoietic precursor cells, and that ablation of PIR in the U937 myelomonocytic cell line or in murine primary hematopoietic precursor cells results in impairment of terminal myeloid differentiation. Gene expression profiling of U937 cells after knockdown of PIR revealed increased expression of genes associated with the early phases of hematopoiesis, in particular, homeobox A (HOXA) genes. Our results suggest that PIR is required for terminal myeloid maturation, and its downregulation may contribute to the differentiation arrest associated with AML. PMID- 20010625 TI - Long-term results of five consecutive trials in childhood acute lymphoblastic leukemia performed by the ALL-BFM study group from 1981 to 2000. AB - Between 1981 and 2000, 6609 children (<18 years of age) were treated in five consecutive trials of the Berlin-Frankfurt-Munster (BFM) study group for childhood acute lymphoblastic leukemia (ALL). Patients were treated in up to 82 centers in Germany, Austria and Switzerland. Probability of 10-year event-free survival (EFS) (survival) improved from 65% (77%) in study ALL-BFM 81 to 78% (85%) in ALL-BFM 95. In parallel to relapse reduction, major efforts focused on reducing acute and late toxicity through advanced risk adaptation of treatment. The major findings derived from these ALL-BFM trials were as follows: (1) preventive cranial radiotherapy could be safely reduced to 12 Gy in T-ALL and high-risk (HR) ALL patients, and eliminated in non- HR non-T-ALL patients, if it was replaced by high-dose and intrathecal (IT) MTX; (2) omission of delayed re intensification severely impaired outcome of low-risk patients; (3) 6-month-less maintenance therapy caused an increase in systemic relapses; (4) slow response to an initial 7-day prednisone window was identified as adverse prognostic factor; (5) condensed induction therapy resulted in significant improvement of outcome; (6) the daunorubicin dose in induction could be safely reduced in low-risk patients and (7) intensification of consolidation/re-intensification treatment led to considerable improvement of outcome in HR patients. PMID- 20010626 TI - AAV-mediated knockdown of peripherin-2 in vivo using miRNA-based hairpins. AB - Gene therapy for inherited retinal degeneration in which expression of a mutant allele has a gain-of-function effect on photoreceptor cells is likely to depend on efficient silencing of the mutated allele. Peripherin-2 (Prph2, also known as peripherin/RDS) is an abundantly expressed photoreceptor-specific gene. In humans, gain-of-function mutations in PRPH2 result in both autosomal dominant retinitis pigmentosa and dominant maculopathies. Gene-silencing strategies for these conditions include RNA interference by short hairpin RNAs (shRNAs). Recent evidence suggests that microRNA (miRNA)-based hairpins may offer a safer and more effective alternative. In this study, we used for the first time a virally transferred miRNA-based hairpin to silence Prph2 in the murine retina. The results show that an miRNA-based shRNA can efficiently and specifically silence Prph2 in vivo as early as 3 weeks after AAV2/8-mediated subretinal delivery, leading to a nearly 50% reduction of photoreceptor cells after 5 weeks. We conclude that miRNA-based hairpins can achieve rapid and robust gene silencing after efficient vector-mediated delivery to the retina. The rationale of using an miRNA-based template to improve the silencing efficiency of a hairpin may prove valuable for allele-specific silencing in which the choice for an RNAi target is limited and offers an alternative therapeutic strategy for the treatment of dominant retinopathies. PMID- 20010627 TI - Dendritic and tumor cell fusions transduced with adenovirus encoding CD40L eradicate B-cell lymphoma and induce a Th17-type response. AB - Fusion of dendritic cells and tumor cells (FCs) constitutes a promising tool for generating an antitumor response because of their capacity to present tumor antigens and provide appropriate costimulatory signals. CD40-CD40L interaction has an important role in the maturation and survival of dendritic cells and provides critical help for T-cell priming. In this study, we sought to improve the effectiveness of FC vaccines in a murine model of B-cell lymphoma by engineering FCs to express CD40L by means of an adenovirus encoding CD40L (Adv CD40L). Before transduction with Adv-CD40L, no CD40L expression was detected in FCs, DCs or tumor cells. The surface expression of CD40L in FC transduced with Adv-CD40L (FC-CD40L) ranged between 50 and 60%. FC-CD40L showed an enhanced expression of CD80, CD86, CD54 and MHC class II molecules and elicited a strong in vitro immune response in a syngeneic mixed lymphocyte reaction. Furthermore, FC-CD40L showed enhanced migration to secondary lymphoid organs. Splenocytes from mice treated with FC-CD40L had a dramatic increase in the production of IL-17, IL 6 and IFN-gamma, compared with controls. Treatment with the FC-CD40L vaccine induced regression of established tumors and increased survival. Our data demonstrate that FC transduced with Adv-CD40L enhances the antitumor effect of FC vaccines in a murine lymphoma model and this is associated with an increased Th17 type immune response. PMID- 20010628 TI - Functional assessment of skeletal muscle in intact mice lacking myostatin by concurrent NMR imaging and spectroscopy. AB - Inhibiting myostatin (mstn) causes spectacular increase in muscle mass, spurring research for therapeutic approaches against neuromuscular disorders. Yet, possible functional deterioration and compromised force production have been reported in isolated muscle of null mstn(-/-) mice. We analyzed vascular and metabolic response to repeated electro-stimulated exercise in vivo in mstn(-/-) mice compared with FVB wild-type controls (WT), using interleaved multi parametric functional nuclear magnetic resonance (NMR) imaging and spectroscopy. At steady-state exercise, specific force of plantar flexion, phosphocreatine consumption measured by phosphorus spectroscopy and maximum perfusion measured by arterial spin-labeled (ASL) NMR imaging were identical in both groups. After exercise, phosphorus spectroscopy revealed reduced oxidative mitochondrial capacity in mstn(-/-), whereas early recovery perfusion was identical and oxygen extraction, estimated from the blood oxygen level-dependent (BOLD) contrast, was decreased when compared with WT. Hyperemia was prolonged, indicating specific regulation of the perfusional response in mstn(-/-) mice. Histology showed an increased proportion of type IIb fibers in hypertrophied muscles, but the distribution of capillary contacts per fiber between oxidative and glycolytic fibers was unaltered in mstn(-/-) compared with WT. These integrated results formed coherent evidence of a congruous, non-pathologic shift toward a more glycolytic metabolism in this model of mstn(-/-). PMID- 20010629 TI - Intramyocardial transplantation of fibroblasts expressing vascular endothelial growth factor attenuates cardiac dysfunction. AB - In this study, we analyzed whether transplantation of cardiac fibroblasts (CFs) expressing vascular endothelial growth factor (VEGF) mitigates cardiac dysfunction after myocardial infarction (MI) in rats. First, we observed that the transgene expression lasts longer (45 vs 7 days) when fibroblasts are used as vectors compared with myoblasts. In a preventive protocol, induction of cardiac neovascularization accompanied by reduction in myocardial scar area was observed when cell transplantation was performed 1 week before ischemia/reperfusion and the animals analyzed 3 weeks later. Finally, the therapeutic efficacy of this approach was tested injecting cells in a fibrin biopolymer, to increase cardiac retention, 24 h post-MI. After 4 weeks, an increase in neovascularization and a decrease in myocardial collagen were observed only in rats that received cells expressing VEGF. Basal indirect or direct hemodynamic measurements showed no differences among the groups whereas under pharmacological stress, only the group that received cells expressing VEGF showed a significant reduction in end diastolic pressure and improvement in stroke volume and cardiac work. These results indicate that transplantation of CFs expressing VEGF using fibrin biopolymer induces neovascularization and attenuates left ventricle fibrosis and cardiac dysfunction in ischemic heart. PMID- 20010631 TI - Aerobic denitrification in permeable Wadden Sea sediments. AB - Permeable or sandy sediments cover the majority of the seafloor on continental shelves worldwide, but little is known about their role in the coastal nitrogen cycle. We investigated the rates and controls of nitrogen loss at a sand flat (Janssand) in the central German Wadden Sea using multiple experimental approaches, including the nitrogen isotope pairing technique in intact core incubations, slurry incubations, a flow-through stirred retention reactor and microsensor measurements. Results indicate that permeable Janssand sediments are characterized by some of the highest potential denitrification rates (> or =0.19 mmol N m(-2) h(-1)) in the marine environment. Moreover, several lines of evidence showed that denitrification occurred under oxic conditions. In intact cores, microsensor measurements showed that the zones of nitrate/nitrite and O(2) consumption overlapped. In slurry incubations conducted with (15)NO(3)(-) enrichment in gas-impermeable bags, denitrification assays revealed that N(2) production occurred at initial O(2) concentrations of up to approximately 90 microM. Initial denitrification rates were not substantially affected by O(2) in surficial (0-4 cm) sediments, whereas rates increased by twofold with O(2) depletion in the at 4-6 cm depth interval. In a well mixed, flow-through stirred retention reactor (FTSRR), (29)N(2) and (30)N(2) were produced and O(2) was consumed simultaneously, as measured online using membrane inlet mass spectrometry. We hypothesize that the observed high denitrification rates in the presence of O(2) may result from the adaptation of denitrifying bacteria to recurrent tidally induced redox oscillations in permeable sediments at Janssand. PMID- 20010630 TI - Hydrography shapes bacterial biogeography of the deep Arctic Ocean. AB - It has been long debated as to whether marine microorganisms have a ubiquitous distribution or patterns of biogeography, but recently a consensus for the existence of microbial biogeography is emerging. However, the factors controlling the distribution of marine bacteria remain poorly understood. In this study, we combine pyrosequencing and traditional Sanger sequencing of the 16S rRNA gene to describe in detail bacterial communities from the deep Arctic Ocean. We targeted three separate water masses, from three oceanic basins and show that bacteria in the Arctic Ocean have a biogeography. The biogeographical distribution of bacteria was explained by the hydrography of the Arctic Ocean and subsequent circulation of its water masses. Overall, this first taxonomic description of deep Arctic bacteria communities revealed an abundant presence of SAR11 (Alphaproteobacteria), SAR406, SAR202 (Chloroflexi) and SAR324 (Deltaproteobacteria) clusters. Within each cluster, the abundance of specific phylotypes significantly varied among water masses. Water masses probably act as physical barriers limiting the dispersal and controlling the diversity of bacteria in the ocean. Consequently, marine microbial biogeography involves more than geographical distances, as it is also dynamically associated with oceanic processes. Our ocean scale study suggests that it is essential to consider the coupling between microbial and physical oceanography to fully understand the diversity and function of marine microbes. PMID- 20010632 TI - Eukarya 18S rRNA gene diversity in the sea surface microlayer: implications for the structure of the neustonic microbial loop. AB - We have previously shown that there is a consistent and reproducible bacterioneuston community in the surface microlayer during a fjord mesocosm experiment. One possible cause of the surface microlayer-specific bacterial community is a surface microlayer-specific protist community selectively grazing on the bacterioneuston. We determined protist community structures using Eukarya 18S rRNA gene denaturing gradient gel electrophoresis (DGGE) and subsequent DGGE band sequencing using DNA samples that were collected from the surface microlayer and subsurface water of the mesocosms. As with bacterial communities, protist community structure was consistently different in the surface microlayer when compared with subsurface water. In particular, the protist community in the surface microlayer was dominated by Cercozoa, which were not detected in the subsurface water, and Ciliophora. PMID- 20010633 TI - Significant roles of bacteriochlorophylla supplemental to chlorophylla in the ocean. AB - Bacteriochlorophylla (BChla)-based phototrophy cannot be ignored in understanding of carbon cycle in the ocean. However, the role of BChla relative to chlorophylla (Chla) along environmental gradients remains unclear. On the basis of the investigations in the Pacific, Indian Oceans and the China Seas, we show that the average contribution of BChla relative to Chla is higher in open-ocean compared with shelf environments, the estimated BChla-based phototrophy could reduce respiratory organic carbon consumption of 2.4% and 5.4% of the primary productions in the shelf and oceanic waters, respectively. This suggests that BChla-based phototrophy has an important role supplemental to Chla-based photosynthesis in the ocean's carbon cycle and would be critical for a marine region to be a sink or a source of atmospheric CO(2). PMID- 20010634 TI - Molecular detection of anammox bacteria in terrestrial ecosystems: distribution and diversity. AB - Anaerobic oxidation of ammonium (anammox) is recognized as an important process in the marine nitrogen cycle yet nothing is known about the distribution, diversity and activity of anammox bacteria in the terrestrial realm. In this study, we report on the detection of anammox sequences of Candidatus 'Brocadia', 'Kuenenia', 'Scalindua' and 'Jettenia' in marshes, lakeshores, a contaminated porous aquifer, permafrost soil, agricultural soil and in samples associated with nitrophilic or nitrogen-fixing plants. This suggests a higher diversity of anammox bacteria in terrestrial than in marine ecosystems and could be a consequence of the larger variety of suitable niches in soils. Anammox bacteria were not ubiquitously present but were only detected in certain soil types and at particular depths, thus reflecting specific ecological requirements. As opposed to marine water column habitats where Candidatus 'Scalindua' dominates anammox guilds, 'Kuenenia' and 'Brocadia' appear to be the most common representatives in terrestrial environments. PMID- 20010635 TI - Molecular analysis of phosphate limitation in Geobacteraceae during the bioremediation of a uranium-contaminated aquifer. AB - Nutrient limitation is an environmental stress that may reduce the effectiveness of bioremediation strategies, especially when the contaminants are organic compounds or when organic compounds are added to promote microbial activities such as metal reduction. Genes indicative of phosphate-limitation were identified by microarray analysis of chemostat cultures of Geobacter sulfureducens. This analysis revealed that genes in the pst-pho operon, which is associated with a high-affinity phosphate uptake system in other microorganisms, had significantly higher transcript abundance under phosphate-limiting conditions, with the genes pstB and phoU upregulated the most. Quantitative PCR analysis of pstB and phoU transcript levels in G. sulfurreducens grown in chemostats demonstrated that the expression of these genes increased when phosphate was removed from the culture medium. Transcripts of pstB and phoU within the subsurface Geobacter species predominating during an in situ uranium-bioremediation field experiment were more abundant than in chemostat cultures of G. sulfurreducens that were not limited for phosphate. Addition of phosphate to incubations of subsurface sediments did not stimulate dissimilatory metal reduction. The added phosphate was rapidly adsorbed onto the sediments. The results demonstrate that Geobacter species can effectively reduce U(VI) even when experiencing suboptimal phosphate concentrations and that increasing phosphate availability with phosphate additions is difficult to achieve because of the high reactivity of this compound. This transcript-based approach developed for diagnosing phosphate limitation should be applicable to assessing the potential need for additional phosphate in other bioremediation processes. PMID- 20010636 TI - Evidence for escape from adaptive conflict? AB - Gene duplication is the primary source of new genes, but the molecular evolutionary mechanisms underlying functional divergence of duplicate genes are not well understood. Des Marais and Rausher argued that data from plant dihydroflavonol-4-reductase (DFR) genes support the model that gene duplication allows the escape from adaptive conflict (EAC) among several functions of a single-copy progenitor gene. As the authors indicated, the key predictions of EAC, in comparison to other models, are that (i) adaptive changes occur in both daughter genes after duplication, and (ii) these adaptive changes must improve ancestral functions. Furthermore, EAC indicates that (iii) the improvement of several ancestral functions is constrained before duplication, although this last point was not explicitly stated. Here we show that contrary to the predictions of EAC, only one of the duplicated DFR lineages exhibited adaptive sequence changes. Owing to the lack of information on enzyme concentrations we question the accuracy of enzyme activity comparisons, and it is thus not clear that any ancestral function has been improved in either lineage. PMID- 20010640 TI - A slippery slope. PMID- 20010641 TI - A question of integrity. PMID- 20010651 TI - Journal club. A biogeochemist ponders muddy molecules and past climates. PMID- 20010654 TI - Plagiarism scandal grows in Iran. PMID- 20010655 TI - Exoplanet claim bites the dust. PMID- 20010656 TI - Culture clash at Australian synchrotron. PMID- 20010657 TI - Primate study halted by US university. PMID- 20010658 TI - 'Killer application' for protein synthesis is retracted. PMID- 20010659 TI - Cattle disease faces total wipeout. PMID- 20010660 TI - Centre turns away from healing herbs. PMID- 20010661 TI - Israel weighs up new funding agency. PMID- 20010662 TI - Copenhagen: the scientists' view. PMID- 20010663 TI - Emerging disease: Looking for trouble. PMID- 20010664 TI - Emissions: Canada should not be isolating itself. PMID- 20010665 TI - Emissions: taking a collaborative lead will work better. PMID- 20010666 TI - Need for religions to promote values of conservation. PMID- 20010667 TI - Time to future-proof plants in storage. PMID- 20010672 TI - Developmental biology: Asymmetry with a twist. PMID- 20010673 TI - Gamma-ray bursts: Magnetism in a cosmic blast. PMID- 20010675 TI - Neuroscience: Excitatory view of a receptor. PMID- 20010676 TI - Earth science: The enigma of D''. PMID- 20010677 TI - Immunology: Dendritic-cell genealogy. PMID- 20010678 TI - Structural biology: Molecular coin slots for urea. PMID- 20010679 TI - Obituary: Qian Xuesen (1911-2009). PMID- 20010680 TI - Systems chemistry: Molecular networks come of age. PMID- 20010681 TI - Encounter and extrusion of an intrahelical lesion by a DNA repair enzyme. AB - How living systems detect the presence of genotoxic damage embedded in a million fold excess of undamaged DNA is an unresolved question in biology. Here we have captured and structurally elucidated a base-excision DNA repair enzyme, MutM, at the stage of initial encounter with a damaged nucleobase, 8-oxoguanine (oxoG), nested within a DNA duplex. Three structures of intrahelical oxoG-encounter complexes are compared with sequence-matched structures containing a normal G base in place of an oxoG lesion. Although the protein-DNA interfaces in the matched complexes differ by only two atoms-those that distinguish oxoG from G their pronounced structural differences indicate that MutM can detect a lesion in DNA even at the earliest stages of encounter. All-atom computer simulations show the pathway by which encounter of the enzyme with the lesion causes extrusion from the DNA duplex, and they elucidate the critical free energy difference between oxoG and G along the extrusion pathway. PMID- 20010682 TI - Ten per cent polarized optical emission from GRB 090102. AB - The nature of the jets and the role of magnetic fields in gamma-ray bursts (GRBs) remains unclear. In a baryon-dominated jet only weak, tangled fields generated in situ through shocks would be present. In an alternative model, jets are threaded with large-scale magnetic fields that originate at the central engine and that accelerate and collimate the material. To distinguish between the models the degree of polarization in early-time emission must be measured; however, previous claims of gamma-ray polarization have been controversial. Here we report that the early optical emission from GRB 090102 was polarized at 10 +/- 1 per cent, indicating the presence of large-scale fields originating in the expanding fireball. If the degree of polarization and its position angle were variable on timescales shorter than our 60-second exposure, then the peak polarization may have been larger than ten per cent. PMID- 20010683 TI - Disordered, quasicrystalline and crystalline phases of densely packed tetrahedra. AB - All hard, convex shapes are conjectured by Ulam to pack more densely than spheres, which have a maximum packing fraction of phi = pi/ radical18 approximately 0.7405. Simple lattice packings of many shapes easily surpass this packing fraction. For regular tetrahedra, this conjecture was shown to be true only very recently; an ordered arrangement was obtained via geometric construction with phi = 0.7786 (ref. 4), which was subsequently compressed numerically to phi = 0.7820 (ref. 5), while compressing with different initial conditions led to phi = 0.8230 (ref. 6). Here we show that tetrahedra pack even more densely, and in a completely unexpected way. Following a conceptually different approach, using thermodynamic computer simulations that allow the system to evolve naturally towards high-density states, we observe that a fluid of hard tetrahedra undergoes a first-order phase transition to a dodecagonal quasicrystal, which can be compressed to a packing fraction of phi = 0.8324. By compressing a crystalline approximant of the quasicrystal, the highest packing fraction we obtain is phi = 0.8503. If quasicrystal formation is suppressed, the system remains disordered, jams and compresses to phi = 0.7858. Jamming and crystallization are both preceded by an entropy-driven transition from a simple fluid of independent tetrahedra to a complex fluid characterized by tetrahedra arranged in densely packed local motifs of pentagonal dipyramids that form a percolating network at the transition. The quasicrystal that we report represents the first example of a quasicrystal formed from hard or non-spherical particles. Our results demonstrate that particle shape and entropy can produce highly complex, ordered structures. PMID- 20010684 TI - Catastrophic flood of the Mediterranean after the Messinian salinity crisis. AB - The Mediterranean Sea became disconnected from the world's oceans and mostly desiccated by evaporation about 5.6 million years ago during the Messinian salinity crisis. The Atlantic waters found a way through the present Gibraltar Strait and rapidly refilled the Mediterranean 5.33 million years ago in an event known as the Zanclean flood. The nature, abruptness and evolution of this flood remain poorly constrained. Borehole and seismic data show incisions over 250 m deep on both sides of the Gibraltar Strait that have previously been attributed to fluvial erosion during the desiccation. Here we show the continuity of this 200-km-long channel across the strait and explain its morphology as the result of erosion by the flooding waters, adopting an incision model validated in mountain rivers. This model in turn allows us to estimate the duration of the flood. Although the available data are limited, our findings suggest that the feedback between water flow and incision in the early stages of flooding imply discharges of about 10(8) m(3) s(-1) (three orders of magnitude larger than the present Amazon River) and incision rates above 0.4 m per day. Although the flood started at low water discharges that may have lasted for up to several thousand years, our results suggest that 90 per cent of the water was transferred in a short period ranging from a few months to two years. This extremely abrupt flood may have involved peak rates of sea level rise in the Mediterranean of more than ten metres per day. PMID- 20010685 TI - Thickness and Clapeyron slope of the post-perovskite boundary. AB - The thicknesses and Clapeyron slopes of mantle phase boundaries strongly influence the seismic detectability of the boundaries and convection in the mantle. The unusually large positive Clapeyron slope found for the boundary between perovskite (Pv) and post-perovskite (pPv) (the 'pPv boundary') would destabilize high-temperature anomalies in the lowermost mantle, in disagreement with the seismic observations. Here we report the thickness of the pPv boundary in (Mg(0.91)Fe(2+)(0.09))SiO(3) and (Mg(0.9)Fe(3+)(0.1))(Al(0.1)Si(0.9))O(3) as determined in a laser-heated diamond-anvil cell under in situ high-pressure (up to 145 GPa), high-temperature (up to 3,000 K) conditions. The measured Clapeyron slope is consistent with the D'' discontinuity. In both systems, however, the pPv boundary thickness increases to 400-600 +/- 100 km, which is substantially greater than the thickness of the D'' discontinuity (<30 km). Although the Fe(2+) buffering effect of ferropericlase could decrease the pPv boundary thickness, the boundary may remain thick in a pyrolitic composition because of the effects of Al and the rapid temperature increase in the D'' layer. The pPv boundary would be particularly thick in regions with an elevated Al content and/or a low Mg/Si ratio, reducing the effects of the large positive Clapeyron slope on the buoyancy of thermal anomalies and stabilizing compositional heterogeneities in the lowermost mantle. If the pPv transition is the source of the D'' discontinuity, regions with sharp discontinuities may require distinct compositions, such as a higher Mg/Si ratio or a lower Al content. PMID- 20010686 TI - Reproductive skew and selection on female ornamentation in social species. AB - Male animals are typically more elaborately ornamented than females. Classic sexual selection theory notes that because sperm are cheaper to produce than eggs, and because males generally compete more intensely for reproductive opportunities and invest less in parental care than females, males can obtain greater fitness benefits from mating multiply. Therefore, sexual selection typically results in male-biased sex differences in secondary sexual characters. This generality has recently been questioned, because in cooperatively breeding vertebrates, the strength of selection on traits used in intrasexual competition for access to mates (sexual selection) or other resources linked to reproduction (social selection) is similar in males and females. Because selection is acting with comparable intensity in both sexes in cooperatively breeding species, the degree of sexual dimorphism in traits used in intrasexual competition should be reduced in cooperative breeders. Here we use the socially diverse African starlings (Sturnidae) to demonstrate that the degree of sexual dimorphism in plumage and body size is reduced in cooperatively breeding species as a result of increased selection on females for traits that increase access to reproductive opportunities, other resources, or higher social status. In cooperative breeders such as these, where there is unequal sharing of reproduction (reproductive skew) among females, and where female dominance rank influences access to mates and other resources, intrasexual competition among females may be intense and ultimately select for female trait elaboration. Selection is thereby acting with different intensities on males and females in cooperatively versus non cooperatively breeding species, and female-female interactions in group-living vertebrates will have important consequences for the evolution of female morphological, physiological and behavioural traits. PMID- 20010687 TI - Ecoenzymatic stoichiometry of microbial organic nutrient acquisition in soil and sediment. AB - Biota can be described in terms of elemental composition, expressed as an atomic ratio of carbon:nitrogen:phosphorus (refs 1-3). The elemental stoichiometry of microoorganisms is fundamental for understanding the production dynamics and biogeochemical cycles of ecosystems because microbial biomass is the trophic base of detrital food webs. Here we show that heterotrophic microbial communities of diverse composition from terrestrial soils and freshwater sediments share a common functional stoichiometry in relation to organic nutrient acquisition. The activities of four enzymes that catalyse the hydrolysis of assimilable products from the principal environmental sources of C, N and P show similar scaling relationships over several orders of magnitude, with a mean ratio for C:N:P activities near 1:1:1 in all habitats. We suggest that these ecoenzymatic ratios reflect the equilibria between the elemental composition of microbial biomass and detrital organic matter and the efficiencies of microbial nutrient assimilation and growth. Because ecoenzymatic activities intersect the stoichiometric and metabolic theories of ecology, they provide a functional measure of the threshold at which control of community metabolism shifts from nutrient to energy flow. PMID- 20010688 TI - Cold-induced silencing by long antisense transcripts of an Arabidopsis Polycomb target. AB - Transcription in eukaryotic genomes generates an extensive array of non-protein coding RNA, the functional significance of which is mostly unknown. We are investigating the link between non-coding RNA and chromatin regulation through analysis of FLC - a regulator of flowering time in Arabidopsis and a target of several chromatin pathways. Here we use an unbiased strategy to characterize non coding transcripts of FLC and show that sense/antisense transcript levels correlate in a range of mutants and treatments, but change independently in cold treated plants. Prolonged cold epigenetically silences FLC in a Polycomb-mediated process called vernalization. Our data indicate that upregulation of long non coding antisense transcripts covering the entire FLC locus may be part of the cold-sensing mechanism. Induction of these antisense transcripts occurs earlier than, and is independent of, other vernalization markers and coincides with a reduction in sense transcription. We show that addition of the FLC antisense promoter sequences to a reporter gene is sufficient to confer cold-induced silencing of the reporter. Our data indicate that cold-induced FLC antisense transcripts have an early role in the epigenetic silencing of FLC, acting to silence FLC transcription transiently. Recruitment of the Polycomb machinery then confers the epigenetic memory. Antisense transcription events originating from 3' ends of genes might be a general mechanism to regulate the corresponding sense transcription in a condition/stage-dependent manner. PMID- 20010689 TI - Long-range oncogenic activation of Igh-c-myc translocations by the Igh 3' regulatory region. AB - B-cell malignancies, such as human Burkitt's lymphoma, often contain translocations that link c-myc or other proto-oncogenes to the immunoglobulin heavy chain locus (IgH, encoded by Igh). The nature of elements that activate oncogenes within such translocations has been a long-standing question. Translocations within Igh involve DNA double-strand breaks initiated either by the RAG1/2 endonuclease during variable, diversity and joining gene segment (V(D)J) recombination, or by activation-induced cytidine deaminase (AID, also known as AICDA) during class switch recombination (CSR). V(D)J recombination in progenitor B (pro-B) cells assembles Igh variable region exons upstream of mu constant region (Cmu) exons, which are the first of several sets of C(H) exons ('C(H) genes') within a C(H) locus that span several hundred kilobases (kb). In mature B cells, CSR deletes Cmu and replaces it with a downstream C(H) gene. An intronic enhancer (iEmu) between the variable region exons and Cmu promotes V(D)J recombination in developing B cells. Furthermore, the Igh 3' regulatory region (Igh3'RR) lies downstream of the C(H) locus and modulates CSR by long-range transcriptional enhancement of C(H) genes. Transgenic mice bearing iEmu or Igh3'RR sequences fused to c-myc are predisposed to B lymphomas, demonstrating that such elements can confer oncogenic c-myc expression. However, in many B-cell lymphomas, Igh-c-myc translocations delete iEmu and place c-myc up to 200 kb upstream of the Igh3'RR. Here we address the oncogenic role of the Igh3'RR by inactivating it in two distinct mouse models for B-cell lymphoma with Igh-c-myc translocations. We show that the Igh3'RR is dispensable for pro-B-cell lymphomas with V(D)J recombination-initiated translocations, but is required for peripheral B-cell lymphomas with CSR-associated translocations. As the Igh3'RR is not required for CSR-associated Igh breaks or Igh-c-myc translocations in peripheral B-cell lymphoma progenitors, we conclude that this regulatory region confers oncogenic activity by long-range and developmental stage-specific activation of translocated c-myc genes. PMID- 20010690 TI - Paradox of mistranslation of serine for alanine caused by AlaRS recognition dilemma. AB - Mistranslation arising from confusion of serine for alanine by alanyl-tRNA synthetases (AlaRSs) has profound functional consequences. Throughout evolution, two editing checkpoints prevent disease-causing mistranslation from confusing glycine or serine for alanine at the active site of AlaRS. In both bacteria and mice, Ser poses a bigger challenge than Gly. One checkpoint is the AlaRS editing centre, and the other is from widely distributed AlaXps-free-standing, genome encoded editing proteins that clear Ser-tRNA(Ala). The paradox of misincorporating both a smaller (glycine) and a larger (serine) amino acid suggests a deep conflict for nature-designed AlaRS. Here we show the chemical basis for this conflict. Nine crystal structures, together with kinetic and mutational analysis, provided snapshots of adenylate formation for each amino acid. An inherent dilemma is posed by constraints of a structural design that pins down the alpha-amino group of the bound amino acid by using an acidic residue. This design, dating back more than 3 billion years, creates a serendipitous interaction with the serine OH that is difficult to avoid. Apparently because no better architecture for the recognition of alanine could be found, the serine misactivation problem was solved through free-standing AlaXps, which appeared contemporaneously with early AlaRSs. The results reveal unconventional problems and solutions arising from the historical design of the protein synthesis machinery. PMID- 20010692 TI - FANCM regulates DNA chain elongation and is stabilized by S-phase checkpoint signalling. AB - FANCM binds and remodels replication fork structures in vitro. We report that in vivo, FANCM controls DNA chain elongation in an ATPase-dependent manner. In the presence of replication inhibitors that do not damage DNA, FANCM counteracts fork movement, possibly by remodelling fork structures. Conversely, through damaged DNA, FANCM promotes replication and recovers stalled forks. Hence, the impact of FANCM on fork progression depends on the underlying hindrance. We further report that signalling through the checkpoint effector kinase Chk1 prevents FANCM from degradation by the proteasome after exposure to DNA damage. FANCM also acts in a feedback loop to stabilize Chk1. We propose that FANCM is a ringmaster in the response to replication stress by physically altering replication fork structures and by providing a tight link to S-phase checkpoint signalling. PMID- 20010693 TI - 53BP1 promotes ATM activity through direct interactions with the MRN complex. AB - The Mre11/Rad50/Nbs1 (MRN) complex has a central function in facilitating activation of the ATM protein kinase at sites of DNA double-strand breaks (DSBs). However, several other factors are also required in human cells for efficient signalling through MRN and ATM, including the tumour suppressor proteins p53 binding protein 1 (53BP1) and BRCA1. In this study, we investigate the functions of these mediator proteins in ATM activation and find that the presence of 53BP1 and BRCA1 can amplify the effects of MRN when interactions between MRN and ATM are compromised. This effect is dependent on a direct interaction between MRN and the tandem breast cancer carboxy-terminal (BRCT) repeats in 53BP1, and is accompanied by hyper-phosphorylation of both Nbs1 and 53BP1. We also find that the BRCT domains of 53BP1 affect the overall structure of 53BP1 multimers and that this structure is important for promoting ATM phosphorylation of substrates as well as for the repair of DNA DSBs in mammalian cells. PMID- 20010694 TI - Modular architecture of Munc13/calmodulin complexes: dual regulation by Ca2+ and possible function in short-term synaptic plasticity. AB - Ca(2+) signalling in neurons through calmodulin (CaM) has a prominent function in regulating synaptic vesicle trafficking, transport, and fusion. Importantly, Ca(2+)-CaM binds a conserved region in the priming proteins Munc13-1 and ubMunc13 2 and thus regulates synaptic neurotransmitter release in neurons in response to residual Ca(2+) signals. We solved the structure of Ca(2+)(4)-CaM in complex with the CaM-binding domain of Munc13-1, which features a novel 1-5-8-26 CaM-binding motif with two separated mobile structural modules, each involving a CaM domain. Photoaffinity labelling data reveal the same modular architecture in the complex with the ubMunc13-2 isoform. The N-module can be dissociated with EGTA to form the half-loaded Munc13/Ca(2+)(2)-CaM complex. The Ca(2+) regulation of these Munc13 isoforms can therefore be explained by the modular nature of the Munc13/Ca(2+)-CaM interactions, where the C-module provides a high-affinity interaction activated at nanomolar [Ca(2+)](i), whereas the N-module acts as a sensor at micromolar [Ca(2+)](i). This Ca(2+)/CaM-binding mode of Munc13 likely constitutes a key molecular correlate of the characteristic Ca(2+)-dependent modulation of short-term synaptic plasticity. PMID- 20010696 TI - Transduction of RNA-directed DNA methylation signals to repressive histone marks in Arabidopsis thaliana. AB - RNA-directed modification of histones is essential for the maintenance of heterochromatin in higher eukaryotes. In plants, cytosine methylation is an additional factor regulating inactive chromatin, but the mechanisms regulating the coexistence of cytosine methylation and repressive histone modification remain obscure. In this study, we analysed the mechanism of gene silencing mediated by MORPHEUS' MOLECULE1 (MOM1) of Arabidopsis thaliana. Transcript profiling revealed that the majority of up-regulated loci in mom1 carry sequences related to transposons and homologous to the 24-nt siRNAs accumulated in wild type plants that are the hallmarks of RNA-directed DNA methylation (RdDM). Analysis of a single-copy gene, SUPPRESSOR OF drm1 drm2 cmt3 (SDC), revealed that mom1 activates SDC with concomitant reduction of di-methylated histone H3 lysine 9 (H3K9me2) at the tandem repeats in the promoter region without changes in siRNA accumulation and cytosine methylation. The reduction of H3K9me2 is not observed in regions flanking the tandem repeats. The results suggest that MOM1 transduces RdDM signals to repressive histone modification in the core region of RdDM. PMID- 20010695 TI - Antagonism of Beclin 1-dependent autophagy by BCL-2 at the endoplasmic reticulum requires NAF-1. AB - In addition to mitochondria, BCL-2 is located at the endoplasmic reticulum (ER) where it is a constituent of several distinct complexes. Here, we identify the BCL-2-interacting protein at the ER, nutrient-deprivation autophagy factor-1 (NAF 1)-a bitopic integral membrane protein whose defective expression underlies the aetiology of the neurodegenerative disorder Wolfram syndrome 2 (WFS2). NAF-1 contains a two iron-two sulphur coordinating domain within its cytosolic region, which is necessary, but not sufficient for interaction with BCL-2. NAF-1 is displaced from BCL-2 by the ER-restricted BH3-only protein BIK and contributes to regulation of BIK-initiated autophagy, but not BIK-dependent activation of caspases. Similar to BCL-2, NAF-1 is found in association with the inositol 1,4,5 triphosphate receptor and is required for BCL-2-mediated depression of ER Ca(2+) stores. During nutrient deprivation as a physiological stimulus of autophagy, BCL 2 is known to function through inhibition of the autophagy effector and tumour suppressor Beclin 1. NAF-1 is required in this pathway for BCL-2 at the ER to functionally antagonize Beclin 1-dependent autophagy. Thus, NAF-1 is a BCL-2 associated co-factor that targets BCL-2 for antagonism of the autophagy pathway at the ER. PMID- 20010697 TI - FOG-1-mediated recruitment of NuRD is required for cell lineage re-enforcement during haematopoiesis. AB - The transcriptional co-factor Friend of GATA1 (FOG-1) has been shown to interact with subunits of the nucleosome remodelling and histone deacetylase (NuRD) complex through a specific motif located at its N-terminus. To test the importance of FOG-1/NuRD interaction for haematopoiesis in vivo, we generated mice with a mutation that specifically disrupts FOG-1/NuRD interaction (FOG 1(R3K5A)). Homozygous FOG-1(R3K5A) mice were found to have splenomegaly, extramedullary erythropoiesis, granulocytosis and thrombocytopaenia secondary to a block in megakaryocyte maturation. FOG-1(R3K5A/R3K5A) megakaryocytes and erythroid progenitors expressed increased levels of GATA2, showing that FOG 1/NuRD interaction is required for the earlier described 'GATA Switch'. In addition, ablation of FOG-1/NuRD interaction led to inappropriate expression of mast cell and eosinophil-specific genes in the megakaryocyte and erythroid lineages. Chromatin immunoprecipitation experiments revealed that the NuRD complex was not properly recruited to a mast cell gene promoter in FOG 1(R3K5A/R3K5A) megakaryocytes, suggesting that FOG-1/NuRD interaction is required for the direct suppression of mast cell gene expression. Taken together, these results underscore the importance of the FOG-1/NuRD interaction for the re enforcement of lineage commitment during erythropoiesis and megakaryopoiesis in vivo. PMID- 20010699 TI - Haplotype-based case-control study on human apurinic/apyrimidinic endonuclease 1/redox effector factor-1 gene and essential hypertension. AB - BACKGROUND: Oxidative DNA damage is involved in the pathophysiology of essential hypertension (EH), which is a multifactorial disorder. Apurinic/apyrimidinic endonuclease 1/redox effector factor-1 (APE1/REF-1) is an essential endonuclease in the base excision repair pathway of oxidatively damaged DNA, in addition to having reducing properties that promote the binding of redox-sensitive transcription factors. Blood pressure in APE1/REF-1-knockout mice is reported to be significantly higher than in wild-type mice. The aim of this study was to investigate the relationship between EH and the human APE1/REF-1 gene through a haplotype-based case-control study using single-nucleotide polymorphisms (SNPs). METHODS: We selected five SNPs in the human APE1/REF-1 gene (rs1760944, rs3136814, rs17111967, rs3136817, and rs1130409), and performed case-control studies in 265 EH patients and 266 age-matched normotensive (NT) subjects. RESULTS: rs17111967 was found to show nonheterogeneity among Japanese subjects. There were no significant differences in the overall distribution of genotypes or alleles for each SNP between EH and NT groups. In the overall distribution of the haplotype-based case-control study constructed based on rs1760944, rs3136817, and rs1130409, the frequency of the G-T-T haplotype was significantly higher in the EH group than in the NT group (2.1% vs. 0.0%, P = 0.001). Multiple logistic regression analysis also revealed significant differences for the G-T-T haplotype, even after adjustment for confounding factors (OR = 8.600, 95% CI: 1.073-68.951, P = 0.043). CONCLUSIONS: Based on the present results, the G-T-T haplotype appears to be a genetic marker of EH, and the APE1/REF-1 gene appears to be a susceptibility gene for EH. PMID- 20010698 TI - Increased skeletal VEGF enhances beta-catenin activity and results in excessively ossified bones. AB - Vascular endothelial growth factor (VEGF) and beta-catenin both act broadly in embryogenesis and adulthood, including in the skeletal and vascular systems. Increased or deregulated activity of these molecules has been linked to cancer and bone-related pathologies. By using novel mouse models to locally increase VEGF levels in the skeleton, we found that embryonic VEGF over-expression in osteo-chondroprogenitors and their progeny largely pheno-copied constitutive beta catenin activation. Adult induction of VEGF in these cell populations dramatically increased bone mass, associated with aberrant vascularization, bone marrow fibrosis and haematological anomalies. Genetic and pharmacological interventions showed that VEGF increased bone mass through a VEGF receptor 2- and phosphatidyl inositol 3-kinase-mediated pathway inducing beta-catenin transcriptional activity in endothelial and osteoblastic cells, likely through modulation of glycogen synthase kinase 3-beta phosphorylation. These insights into the actions of VEGF in the bone and marrow environment underscore its power as pleiotropic bone anabolic agent but also warn for caution in its therapeutic use. Moreover, the finding that VEGF can modulate beta-catenin activity may have widespread physiological and clinical ramifications. PMID- 20010700 TI - Noninvasive brachial-ankle pulse wave velocity in hypertensive patients with left ventricular hypertrophy. AB - BACKGROUND: The elevation of left ventricular filling pressure (LVFP) could be an important prognostic factor in patients with hypertension. We hypothesized that noninvasive brachial-ankle pulse wave velocity (baPWV) is associated with increased LVFP in hypertensive patients with LV hypertrophy (LVH). METHODS: We enrolled patients with well-controlled hypertension for more than 1 year with LV ejection fraction (LVEF) > or = 55%, and LVH. The relationship between Doppler echocardiographic parameters of LVFP and baPWV with B-type natriuretic peptide (BNP) was also evaluated. RESULTS: A total of 62 patients were enrolled (31 patients with E/E(a) >15 and 31 patients with E/Ea < or = 15) and the baPWV of the E/Ea >15 group was significantly increased compared to the E/Ea < or = 15 group (1,664.3 +/- 270.5 vs. 1,381.9 +/- 159.1 cm/s, P < 0.01). The baPWV showed better correlation with E/Ea (r = 0.69, P < 0.01) than the BNP (r = 0.47, P < 0.01). A multivariate linear regression model showed that only baPWV was significantly correlated with E/E(a), and that the association was independent of other factors. The area under the receiver-operating characteristic (ROC) curve of baPWV for the detection of elevated LVFP (E/Ea >15) was 0.79 (P < 0.01) and the optimal cutoff point of 1,440 cm/s produced 75% sensitivity and 62% specificity (the positive and negative predictive values were 68 and 71%, respectively). CONCLUSIONS: In this study, we have demonstrated that elevated baPWV is associated with noninvasive markers of increased LVFP in hypertensive LVH patients with preserved LV systolic function. PMID- 20010701 TI - Hospitalization costs associated with hypertension as a secondary diagnosis among insured patients aged 18-64 years. AB - BACKGROUND: We estimated the hospitalization costs associated with hypertension as a secondary diagnosis among insured adults aged 18-64 years by using data from 2005 MarketScan Commercial Claims and Encounters (CCAE) inpatient admissions. METHODS: We analyzed costs for four patient groups (N = 455,944): (i) all selected patients; (ii) patients with the primary diagnosis of ischemic heart disease (IHD); (iii) patients with the primary diagnosis of cerebrovascular disease; and (iv) patients with neither IHD nor cerebrovascular disease as the primary diagnosis. We conducted propensity score matching to control possible bias in cost estimates due to sample selections and estimated the costs of hypertension by using a regression model on the matched populations that controlled for subjects' age, sex, length of hospital stay, Charlson comorbidity index (CCI), region of residence, and urbanization of residence. RESULTS: For all patients with hypertension as a secondary diagnosis, the estimated average annual hospitalization cost per patient was $21,094, of which $2,734 (13%; P < 0.01) was associated with hypertension. The estimated average costs were $31,106 for patients with a primary diagnosis of IHD, $17,298 for those with a primary diagnosis of cerebrovascular disease, and $18,693 for those without a primary diagnosis of IHD or cerebrovascular disease; hypertension-associated costs for these patients were $3,540 (11.4%; P < 0.01), $1,133 (6.5%; P < 0.01), and $2,254 (12.1%; P < 0.01), respectively. CONCLUSIONS: Hypertension-associated hospitalization costs are substantial among insured US patients aged 18-64 years with hypertension as a secondary diagnosis and suggest a need for cost-effective programs to prevent, manage, and control hypertension. PMID- 20010702 TI - Neurocircuitry: a window into the networks underlying neuropsychiatric disease. PMID- 20010703 TI - The role of the ventral pallidum in psychiatric disorders. PMID- 20010704 TI - Exploring the molecular basis of addiction: drug-induced neuroadaptations. PMID- 20010705 TI - The emerging role of integrins in neuropsychiatric disorders. PMID- 20010706 TI - Imaging advances in developing new medications for addiction. PMID- 20010707 TI - Sex-related functional asymmetry in the limbic brain. PMID- 20010708 TI - Dissecting the neural circuitry of addiction and psychiatric disease with optogenetics. PMID- 20010709 TI - Neurocartography. PMID- 20010710 TI - Modafinil: an anti-relapse medication. PMID- 20010711 TI - Trick or treat? neurodevelopmental consequences of pharmacotherapy for affective disorders. PMID- 20010712 TI - Ligand functional selectivity advances our understanding of drug mechanisms and drug discovery. PMID- 20010713 TI - Induced pluripotent stem (iPS) cells and their future in psychiatry. PMID- 20010714 TI - Advances in PET analyses of stress and dopamine. PMID- 20010715 TI - Imaging and genetics advances in understanding geriatric depression. PMID- 20010716 TI - Neuronal signaling pathways: genetic insights into the pathophysiology of major mental illness. PMID- 20010717 TI - Antidepressants, age, and neuroprogenitors. PMID- 20010718 TI - Normalizing drug-induced neuronal plasticity in nucleus accumbens weakens enduring drug-seeking behavior. PMID- 20010719 TI - Orexin/hypocretin in psychiatric disorders: present state of knowledge and future potential. PMID- 20010720 TI - New vaccine development for chronic brain disease. PMID- 20010721 TI - Convergent Functional Genomics: what we have learned and can learn about genes, pathways, and mechanisms. PMID- 20010722 TI - Quantification of beta-human papillomavirus DNA by real-time PCR. AB - Quantitative PCR with hybridization probes allows the reliable quantification of viral DNA sequences in clinical samples with a dynamic range and sensitivity that cannot be achieved with other methods. The technical background for the establishment of protocols is described and established protocols are presented to estimate the viral load per cell of frequently occurring betapapillomaviruses (HPV5, -8, -15, -20, -23, -24, -36 and -38) in skin tumors, healthy skin and hair bulbs. This approach accurately adjusts dilution series of reference DNA of different viral types relative to pUC18, which is crucial for comparative analyses and for interlaboratory standardization. The type-specific determination of beta-HPV DNA loads is an important research tool toward discrimination between low-level persistence and activated possibly pathologically relevant infections. The analysis of 24 samples, starting with DNA extraction and followed by HPV typing and quantification of-on average-three of the described HPV types takes about 2 d. PMID- 20010723 TI - A simple slot blot for the detection of virtually all subtypes of the influenza A viral hemagglutinins using universal antibodies targeting the fusion peptide. AB - The fusion peptide of influenza virus hemagglutinin (HA) has a critical role in mediating the entry of the virus into the cells and is also the only universally conserved sequence in the HAs of all strains of influenza A and influenza B viruses. Therefore, it could be an attractive target for new vaccine development and a potency marker for existing influenza vaccines. The fusion peptide epitope is hidden inside the HA proteins, making it inaccessible for quantitative antibody binding. Our simple slot blot protocol highlights pre-treatment of HA samples with moderate concentrations of denaturant to maximally expose the fusion peptide on the protein surface, followed by detection using universal antibodies targeting the fusion peptide. The method is highly reliable, inexpensive and easy to follow. The entire procedure takes only 5 h and can be applied to the quantitative determination of virtually all influenza viral HAs using a single antibody targeting the fusion peptide. PMID- 20010724 TI - Methods for quantifying simple gravity sensing in Drosophila melanogaster. AB - Perception of gravity is essential for animals: most animals possess specific sense organs to detect the direction of the gravitational force. Little is known, however, about the molecular and neural mechanisms underlying their behavioral responses to gravity. Drosophila melanogaster, having a rather simple nervous system and a large variety of molecular genetic tools available, serves as an ideal model for analyzing the mechanisms underlying gravity sensing. Here we describe an assay to measure simple gravity responses of flies behaviorally. This method can be applied for screening genetic mutants of gravity perception. Furthermore, in combination with recent genetic techniques to silence or activate selective sets of neurons, it serves as a powerful tool to systematically identify neural substrates required for the proper behavioral responses to gravity. The assay requires 10 min to perform, and two experiments can be performed simultaneously, enabling 12 experiments per hour. PMID- 20010725 TI - Protocol for quantifying sound-sensing ability of Drosophila melanogaster. AB - Hearing is an important sensory modality for most animals to detect sound signals as they mate, look for food or fend off prey. Despite its critical role in numerous innate behaviors, relatively little is known about how the sensory information regarding the movement of air particles is detected, processed and integrated in the brain. Drosophila melanogaster, with a rather simple nervous system and the large variety of molecular and genetic tools available for its study, is an ideal model organism for dissecting the mechanisms underlying sound sensing. Here we describe assays to measure sound responses of flies behaviorally. Although this method was originally developed for mutant screening, it can also be combined with recent genetic techniques to analyze functions of the identified neural circuits by silencing or activating select sets of neurons. This assay requires approximately 15 min for an experiment and 1.5 h for subsequent analyses. PMID- 20010726 TI - Serum 25-hydroxyvitamin d concentration, life factors and obesity in Mexican children. AB - Although current evidence emphasizes a high prevalence of vitamin D deficiency and an inverse association between serum 25-hydroxyvitamin D (25-OHD) concentration and obesity, no studies have been conducted in Mexican children. The objective was to determine the prevalence of vitamin D deficiency and its association with obesity and lifestyle factors in a sample of school-aged Mexican children. A cross-sectional study of 99 obese and 99 nonobese 6-12 year-old children, skin phototypes III-V, from six public schools was conducted during summer at latitude 25 degrees 40', in northeastern Mexico. Anthropometric measurements were determined. Serum 25-OHD was measured by immunoluminometric direct assay. Consumption of foods rich in vitamin D, sunscreen use and vitamin consumption were assessed through applied questionnaires. 62.1% of the subjects had insufficiency of 25-OHD (21-29 ng/ml) and 20.2% had deficiency (<20 ng/ml). Obese subjects (BMI >or=95th percentile for age and gender) had significantly lower concentration of 25-OHD than nonobese. Predictors of 25-OHD concentration were, in order of significance: percentage of body fat, BMI, triceps skin fold, and waist circumference (WC). A significantly higher rate of 25-OHD deficiency was observed in children with inadequate milk/yoghurt consumption, but no difference was found for other foods, physical activity (PA) or screen-time. In a multivariate model, being obese was significantly associated with the risk of 25 OHD deficiency, after adjustment for PA, screen-time, skin phototype, ingestion of milk/yoghurt, fish, cheese, and carbonated beverages. A high prevalence of vitamin D deficiency and an inverse association between serum 25-OHD concentration and obesity was found. PMID- 20010727 TI - Lack of evidence for the role of human adenovirus-36 in obesity in a European cohort. AB - Adenovirus infection has been shown to increase adiposity in chickens, mice, and nonhuman primates. Adenovirus type 36 (Ad-36) DNA was detected in adipose tissues in these animal trials. In the United States, Ad-36 significantly correlates with obesity as illustrated by an Ad-36 seroprevalence of 30% in obese individuals and 11% in nonobese individuals. We investigated the possibility of a similar correlation of Ad-36 in Dutch and Belgian persons. In total, 509 serum samples were analyzed for Ad-36 antibodies using a serum neutralization assay. In addition, PCR was used to detect adenoviral DNA in visceral adipose tissue of 31 severely obese surgical patients. Our results indicated an overall Ad-36 seroprevalence of 5.5% increasing with age. BMI of Ad-36 seropositive humans was not significantly different from seronegative humans. No adenoviral DNA could be found using PCR on visceral adipose tissue. In conclusion, this first Ad-36 study in the Netherlands and in Belgium indicates that Ad-36 does not play a role as a direct cause of BMI increase and obesity in humans in Western Europe. PMID- 20010728 TI - A national survey of oral and maxillofacial surgeons' attitudes towards the treatment and dental rehabilitation of oral cancer patients. AB - AIMS: To investigate the attitudes of maxillofacial surgeons in the treatment and dental rehabilitation of oral cancer patients in the UK. MATERIAL AND METHODS: The survey was conducted by postal questionnaires with 17 close-ended questions. A total of 229 questionnaires were sent to members of the British Association of Oral and Maxillofacial Surgeons over a one week period. A follow-up was sent if a reply was not received within 12 weeks. These results were compared to a similar study that was carried out approximately 15 years ago. RESULTS: The response rate was 65.5% (150/229). Overall 62% of respondents (92/150) carried out maxillary resections, which represents a decline of 23% on the previous study. There has been an increase in surgeons reconstructing the maxillary defect from 38% in the 1995 study to 91% in the present study. Ninety-eight percent of respondents had their patients seen in a multidisciplinary team (MDT) clinic, but in only 30% of the cases was a restorative dentist present on these clinics. There has been an improvement in the accessibility of a restorative dentist for this patient cohort, from 65% to 90%. The use of implants for dental rehabilitation post cancer surgery has increased from 43% to 93%. CONCLUSION: This study highlights the changes in the dental and oral rehabilitation of patients undergoing resective surgery for oral cancer and especially those undergoing a maxillectomy procedure. It illustrates the increased use of implants for post-surgery rehabilitation and shows the different trends in which these implants are placed. An important aspect of this study is the input of the dental team. Current national guidelines state that a consultant restorative dentist needs to be a member of the MDT; this survey shows that this was the case in only 30% of responses. PMID- 20010729 TI - Patients' knowledge and views about the effects of smoking on their mouths and the involvement of their dentists in smoking cessation activities. AB - BACKGROUND: Smoking is correlated with a large number of oral conditions such as tooth staining and bad breath, periodontal diseases, impaired healing of wounds, precancer and oral cancer. These effects are often visible and in the early stages they are reversible after cessation of smoking. Dentists, as part of the health profession, are frequently in contact with the general population and there is evidence that they are as effective in providing smoking cessation counselling as any other healthcare group. AIMS AND METHODS: Patients' knowledge of the effects of smoking and their attitudes towards the role of dentists in smoking cessation activities were analysed via a self-completing questionnaire and compared depending on their smoking status (smokers and non-smokers). RESULTS: The results show that patients hold very positive attitudes towards dentists' role in smoking cessation. The results also show that although patients have a good knowledge of the effects of smoking on general health, smokers are significantly less aware of the relationship between smoking and gum disease and on wound healing. CONCLUSIONS: Dentists should inform their patients about the oral effects of smoking and strongly advise them not to smoke, especially in patients diagnosed with periodontal disease and requiring surgical procedures. PMID- 20010730 TI - To engage or not to engage--a modern representative's dilemma. PMID- 20010731 TI - Falling below. PMID- 20010732 TI - Help to quit. PMID- 20010733 TI - Tuberculosis diagnosis. PMID- 20010734 TI - Not recommended. PMID- 20010735 TI - Jumping the gun. PMID- 20010736 TI - Voicing support. PMID- 20010737 TI - Liquorice alert. PMID- 20010738 TI - Warm saline rinses. PMID- 20010739 TI - Who will pay? PMID- 20010749 TI - The obsolescence of formocresol. AB - Concern has existed for almost ten years regarding the safety and efficacy of formaldehyde-based medicaments like formocresol in dentistry. Formocresol has been shown to be therapeutically outdated for decades. While the use of formocresol around the world continues to drop, it is still utilised in alarmingly high rates, an age-old bias that is unsubstantiated by overall academic research. Formaldehyde remains a genotoxic and carcinogenic problem worldwide. The most recent articles are discussed in the light of the need to abandon formocresol. PMID- 20010750 TI - Dental therapy in the United Kingdom: part 4. Teamwork--is it working for dental therapists? AB - OBJECTIVES: To determine whether practising dental therapists, including dually qualified hygienist/therapists, considered themselves to be part of the clinical team and whether clinical work referred to them met with their expectations. METHODS: A postal survey enquired about work experiences of UK dental therapists, as previously described earlier in the series. RESULTS: While they certainly considered themselves to be part of the clinical team, the majority of respondents did not feel 'fully utilised'. Seventy percent of respondents felt that the dentist had more patients that could be referred and 55% thought that they could do more extensive work. There was concern that dentists lacked awareness of therapists' clinical potential, although some respondents highlighted very positive experiences in practice. CONCLUSIONS: Dental therapists feel that they are part of the clinical team but consider that their skills are not fully utilised in many cases. There is scope for raising awareness among dentists regarding the therapists' clinical potential as well as sharing ideas for good working practice both within individual clinical settings and between different practices. PMID- 20010757 TI - A new dental school in Cairns. AB - Starting a new school is a challenge, especially in the present economic climate, but that is the situation facing several schools in Australia. Due to the shortage of dental manpower, especially in the more remote areas of Australia, the Federal Government has provided funds to increase the number of dentists in training. One of the areas chosen for a new dental school was Cairns, Northern Queensland and the administration of the new school is under the auspices of the James Cook University (JCU). The Main JCU campus is at Townsville which is 340 km (212 miles) away and about four hours' driving distance (Fig. 1). The Faculty of Medicine, Health and Molecular Sciences offers many health professional programs at both campuses, including Medicine, Physiotherapy and Pharmacy at the Townsville Campus and Dentistry and Nursing in Cairns (Fig. 2). PMID- 20010774 TI - A Himalayan dental adventure! AB - Shekha Bhuva, a Periodontology MClinDent specialist trainee from Guy's Dental Hospital, was lucky enough to take part in a once-in-a-lifetime trip to Nepal as a volunteer dentist. Here she talks about her experiences. PMID- 20010779 TI - Does therapeutic angiogenesis overcome CKD? PMID- 20010780 TI - Remote control of brain angiotensin II levels by angiotensin receptor blockers. PMID- 20010781 TI - Signal transduction of the (pro)renin receptor as a novel therapeutic target for preventing end-organ damage. AB - The (pro)renin receptor ((P)RR) not only represents a novel component of the renin-angiotensin system but is also a promising novel drug target because of its crucial involvement in the pathogenesis of renal and cardiac end-organ damage. This review discusses the signal transduction of the (P)RR with its adapter protein promyelocytic zinc-finger protein, the impact of this receptor, especially on cardiovascular disease, and its putative interaction with renin inhibitors such as aliskiren. Furthermore, the increasing complexity regarding the cellular function of the (P)RR is addressed, which arises by the intimate link with proton pumps and the phosphatase PRL-1, as well as by the presence of different subcellular localizations and of a soluble isoform of the (P)RR. Finally, the rationale and strategy for the development of small-molecule antagonists of the (P)RR, called renin/prorenin receptor blockers, are presented. PMID- 20010782 TI - Influence of ghrelin gene polymorphisms on hypertension and atherosclerotic disease. AB - Ghrelin is involved in several metabolic and cardiovascular processes. Recent evidence suggests its involvement in blood pressure regulation and hypertension. The aim of the study was to determine associations of single-nucleotide polymorphisms (SNPs) and haplotypes of the ghrelin gene (GHRL) with hypertension and atherosclerotic disease. Six GHRL SNPs (rs27647, rs26802, rs34911341, rs696217, rs4684677 and a -473G/A (with no assigned rsID)) were investigated in a sample of 1143 hypertensive subjects and 1489 controls of Caucasian origin. Both single-locus and haplotype association analyses were performed. In single-locus analyses, only the non-synonymous rs34911341 was associated with hypertension (odds ratio (OR)=1.95 (95% confidence interval (CI): 1.26-3.02), P=0.003). Six common haplotypes with frequency >1% were inferred from the studied GHRL SNPs, and their frequency distribution was significantly different between hypertensive subjects and controls (chi(2)=12.96 with 5 d.f. (degree of freedom), P=0.024). The effect of rs26802 was found to be significantly (P=0.017) modulated by other GHRL SNPs, as its C allele conferred either an increased risk (OR=1.30 (1.08 1.57), P=0.005) or a decreased risk (OR=0.50 (0.23-1.06), P=0.07) of hypertension according to the two different haplotypes on which it can be found. No association of GHRL SNPs or haplotypes with atherosclerotic disease was observed. In conclusion, we observed statistical evidence for association between GHRL SNPs and risk of hypertension. PMID- 20010783 TI - Necroptosis, necrosis and secondary necrosis converge on similar cellular disintegration features. AB - Necroptosis, necrosis and secondary necrosis following apoptosis represent different modes of cell death that eventually result in similar cellular morphology including rounding of the cell, cytoplasmic swelling, rupture of the plasma membrane and spilling of the intracellular content. Subcellular events during tumor necrosis factor (TNF)-induced necroptosis, H(2)O(2)-induced necrosis and anti-Fas-induced secondary necrosis were studied using high-resolution time lapse microscopy. The cellular disintegration phase of the three types of necrosis is characterized by an identical sequence of subcellular events, including oxidative burst, mitochondrial membrane hyperpolarization, lysosomal membrane permeabilization and plasma membrane permeabilization, although with different kinetics. H(2)O(2)-induced necrosis starts immediately by lysosomal permeabilization. In contrast, during TNF-mediated necroptosis and anti-Fas induced secondary necrosis, this is a late event preceded by a defined signaling phase. TNF-induced necroptosis depends on receptor-interacting protein-1 kinase, mitochondrial complex I and cytosolic phospholipase A(2) activities, whereas H(2)O(2)-induced necrosis requires iron-dependent Fenton reactions. PMID- 20010785 TI - An essential role for p73 in regulating mitotic cell death. AB - The molecular mechanisms regulating cell death during mitosis are poorly understood. We show here a critical role for p73, but not p53, in regulating mitotic cell death induced by various means. Prolonged mitotic arrest and the activation of spindle checkpoint are required for mitotic death, which occurs before mitotic exit and which can be ameliorated by accelerated mitotic exit. Absence or silencing of p73 expression abrogated mitotic death without accelerating mitotic exit, and was independent of BubR1 and Mad2, the loss of which promotes mitotic exit. However, the absence of p73 reduced mitotic death by compromising the expression of the proapoptotic BH3-only protein Bim and thereby affecting cytochrome c release and caspase activation. p73 was found to induce bim expression through direct binding to regulatory elements in intron 1. Congruently, mitotic cell death was rescued to similar extents by silencing either bim or p73 expression. Taken together, the data show an important role for the p73-Bim axis in regulating cell death during mitosis that is independent of p53. PMID- 20010786 TI - Alcohol dehydrogenase-2 and aldehyde dehydrogenase-2 genotypes, alcohol drinking and the risk for esophageal cancer in a Chinese population. AB - To investigate the relationship among alcohol dehydrogenase-2 (ADH2) and aldehyde dehydrogenase-2 (ALDH2) genetic polymorphisms, alcohol consumption and the susceptibility to esophageal cancer in a Chinese population, we conducted a case control study with 221 cases and 191 population-based controls in the Taixing city of Jiangsu Province of China. ADH2 and ALDH2 genotypes were examined using PCR and denaturing high-performance liquid chromatography. Alcohol drinkers with the ALDH2 A allele showed a significantly increased risk of esophageal cancer compared with drinkers with the ALDH2 G/G genotype (odds ratio (OR)=3.08, 95% confidence interval (CI): 1.65-5.78) or nondrinkers with any genotype (OR=3.05, 95% CI: 1.49-6.25). Drinkers with the ALDH2 A allele and a cumulative amount of alcohol consumption > or =2.5 (kg * years) were at a significantly higher risk of developing esophageal cancer (OR=11.93, 95% CI: 3.17-44.90) compared with individuals with ALDH2 G/G genotypes and a cumulative amount of alcohol consumption <2.5 (kg * years). A dose-dependent positive result was found between cumulative amount of alcohol consumption and risk of esophageal cancer in individuals carrying the ALDH2 A allele (P=0.023) and the homozygous ALDH2 G allele (P=0.047). Compared with individuals carrying both ALDH2 G/G and ADH2 A/A alleles and with a cumulative amount of alcohol consumption <2.5 (kg * years), drinkers carrying both ALDH2 A and ADH2 G alleles and with a cumulative amount of alcohol consumption > or =2.5 (kg * years) showed a significantly elevated risk of esophageal cancer (OR=53.15, 95% CI: 4.24-666.84). This result suggests that to help lower their risk for esophageal cancer, persons carrying the ALDH2 A allele should be encouraged to reduce their consumption of alcoholic beverages. PMID- 20010787 TI - Proteasomes in immune cells: more than peptide producers? AB - When cells are stimulated with pro-inflammatory cytokines, most of their constitutively expressed proteasomes are replaced with immunoproteasomes, which increase the production of peptides for presentation on MHC class I molecules. In addition, cortical thymic epithelial cells selectively express a type of proteasome known as the thymoproteasome that is required for the positive selection of thymocytes. Here, we discuss how these specialized types of proteasome shape the T cell receptor repertoire of cytotoxic T lymphocytes and propose that immunoproteasomes have functions, in addition to antigen processing, that influence cytokine production and T cell differentiation, survival and function. We also discuss how inhibitors of immunoproteasomes can suppress undesired T cell responses in autoimmune diseases. PMID- 20010789 TI - Imaging chorioretinal vascular disease. AB - Since its first description more than 40 years ago, fluorescein angiography had a crucial role in the diagnosis and management of chorioretinal vascular disorders such as neovascular age-related macular degeneration. Although fluorescein angiography permits visualization of the retinal microcirculation in exquisite detail, visualization of the choroidal circulation is more limited. Moreover, fluorescein angiography provides only minimal information regarding the functional consequences of vascular disease and allows, at best, only semi quantitative assessment of retinal thickness. In recent years, the development of other chorioretinal imaging modalities, such as indocyanine green angiography, fundus autofluorescence, and optical coherence tomography (OCT), has addressed many of these issues. In particular, OCT has become an integral tool for vitreoretinal specialists as it allows high-resolution cross-sectional images of the neurosensory retina to be obtained in a non-invasive manner. The latest generation of commercial OCT technology-spectral domain OCT-offers high-speed scanning that allows complete coverage of the macular area, generation of three dimensional retinal reconstructions, and precise image registration for inter visit comparisons. The high speed of spectral domain OCT also facilitates B-scan averaging, which reduces speckle noise artefact and allows unparalleled visualization of the outer retina and choroid. In the near future, further advances in OCT technology (eg Doppler OCT) are likely to dramatically enhance the diagnosis and management of patients with chorioretinal vascular disease. PMID- 20010790 TI - Ab-externo cyclodialysis enhanced trabeculectomy for intractable post-penetrating keratoplasty glaucoma. AB - PURPOSE: To evaluate the efficacy of cyclodialysis-enhanced Mitomycin c (MMC) trabeculectomy in post-keratoplasty glaucoma. DESIGN: Prospective, non comparative, interventional case series. METHODS: A total of 45 eyes of 45 consecutive patients with refractory glaucoma after penetrating glaucoma underwent a cyclodialysis-augmented trabeculectomy with MMC. The visual acuity, intraocular pressure (IOP), corneal clarity, and graft failure were evaluated over a minimum follow-up of 2 years. RESULTS: The mean age of the patients was 55.4+/-9.4 years. The cyclodialysis-augmented MMC trabeculectomy procedure resulted in a mean reduction of IOP from 38.9+/-3.9 mmHg (95% confidence interval (CI) 35.9-42.2) at baseline to 11.3+/-2.8 mmHg (95% CI 9.5-12.9), at final follow up (P=0.002). Blebs were avascular, diffuse, and extended posteriorly. Postoperative UBMs revealed the presence of a cyclodialysis with a small associated suprachoroidal effusion and a subconjunctival filtering bleb in all cases. CONCLUSIONS: Cyclodialysis-augmented MMC trabeculectomy provides a safe and effective method of lowering IOP in intractable glaucoma following penetrating keratoplasty, without compromising the corneal graft survival and clarity. PMID- 20010791 TI - Oxygen-induced retinopathy: a model for vascular pathology in the retina. AB - Ischaemic vascular disease in the retina may either leave retina permanently ischaemic with slow degradation of vision, or alternatively lead to proliferative vascular disease, which can also destroy vision. To investigate the molecular and cellular mechanisms that contribute to this pathology a mouse model has been studied extensively. The model is based on the exposure of mouse pups to hyperoxia during a phase when their retinal vasculature is still developing. This leads to capillary depletion, and upon return to room air, results in retinal ischaemia and proliferative vascular disease in the retinal vasculature (oxygen induced retinopathy (OIR)). Numerous studies using this OIR model have revealed that the regulation of angiogenic factors and the influence of inflammatory cells play a pivotal role in the vascular pathogenesis. It has also been demonstrated in the OIR model that proliferative vascular disease is not the only possible outcome of ischaemia-induced angiogenesis in the retina, but that ischaemic areas in the retina can be revascularised with healthy blood vessels. Therefore, understanding the factors that control the balance between pathological and healthy angiogenesis in the OIR model may have important implications for human retinal ischaemic disease. PMID- 20010788 TI - The immune response during acute HIV-1 infection: clues for vaccine development. AB - The early immune response to HIV-1 infection is likely to be an important factor in determining the clinical course of disease. Recent data indicate that the HIV 1 quasispecies that arise following a mucosal infection are usually derived from a single transmitted virus. Moreover, the finding that the first effective immune responses drive the selection of virus escape mutations provides insight into the earliest immune responses against the transmitted virus and their contributions to the control of acute viraemia. Strong innate and adaptive immune responses occur subsequently but they are too late to eliminate the infection. In this Review, we discuss recent studies on the kinetics and quality of early immune responses to HIV-1 and their implications for developing a successful preventive HIV-1 vaccine. PMID- 20010792 TI - Ultrasound biomicroscopic findings of conventional and sutureless sclerotomy sites after 20-, 23-, and 25-G pars plana vitrectomy. AB - PURPOSE: We examined sclerotomies that were performed using 20-G systems and non stitch 23-G systems, one-step (23/1) and two-step procedures (23/2), as well as one-step 25-G systems for pars plana vitrectomy (PPV) with ultrasound biomicroscopy (UBM) to assess subconjunctival bleb formation, vitreous incarceration, and wound closure. METHODS: This study is a prospective, non randomized, monocentre study comprising 40 patients; we examined sclerotomies using UBM after 20-G, 23/1-G, 23/2-G, and 25-G PPV in 10 patients each on days 1 and 30 postoperatively. RESULTS: Subconjunctival bleb formation was not detected in any of the eyes on the first day after surgery in the 20-G and 23/2-G groups and was observed in 5 (50%, 25 G) and 2 eyes (20%, 23/1 G). By day 30 postoperatively, all blebs had spontaneously resolved. The sclerotomy tunnel could be detected by UBM at day 1 postoperatively in 10 (100%, 20 G), 9 (90%, 25 G), 7 (70%, 23/1 G), and 7 eyes (70%, 23/2 G). Thirty days postoperatively, it was present in 10 (100%, 20 G), 1 (10%, 25 G), 3 (30%, 23/1 G), and 1 (10%, 23/2 G) eye. Vitreous incarceration occurred in 2 (20%, 20 G), 7 (70%, 25 G), 4 (40%, 23/1 G), and 2 (20%, 23/2 G) eyes. CONCLUSIONS: Anatomical closure of sclerotomies without subconjunctival bleb formation as seen on UBM was achieved at day 1 after 23/2-G and 20-G PPV and after 30 days postoperatively in all other non-stitch PPV techniques. Vitreous incarceration is common after 25-G PPV. Non stitch sclerotomies can frequently no longer be detected by UBM 30 days postoperatively. PMID- 20010793 TI - The association between lumican gene polymorphisms and high myopia. AB - PURPOSES: Lumican (LUM) is one of the major extracellular matrix components of the sclera. Increasing evidence suggests that changes in the structure and composition of the sclera are major factors in regulating scleral integrity and axial elongation of the eye, as in myopia. PATIENTS AND METHODS: Patients (n=182; age range, 17-24 years) were with a myopic spherical equivalent (SE)>6.5 diopters (D) and the control group comprised individuals (n=78; age range, 17-25 years) were with a myopic SE<0.5 D. The DNA fragments were separated by horizontal electrophoresis on 3% agarose gels. The forward primer was labelled with a 5' FAM and the reaction products were detected using a 3100 Genetic Analyzer. RESULTS: The polymorphisms detected in this study were LUMc.601, LUM-59, LUM-628, and LUM 1554. Moreover, the haplotype distributions of Ht1 (C/A/CC/T), Ht2 (C/A/--/T), Ht3 (T/A/CC/C), Ht4 (T/--/CC/T), Ht5 (T/--/CC/C), and Ht6 (T/--/--/C) of these polymorphisms were compared between the two groups. The haplotype frequencies of Ht1, Ht2, Ht5, and Ht6 differed significantly between the two groups (P=2.08x10( 5), odds ratio (OR): 2.19, 95% confidence interval (CI): 1.52-3.15; P=2.2x10(-5), OR: 0.39, 95% CI: 0.25-0.61; P=2.7x10(-5), OR: 0.36, 95% CI: 0.22-0.59; P=3.7x10( 5), OR: 4.71, 95% CI: 2.12-10.5, respectively). CONCLUSIONS: These observations suggest that the four polymorphisms of the LUM promoter contribute to the pathogenesis of high myopia. Understanding the functions of LUM in myopia helps us design new methods in treating and preventing myopia. PMID- 20010794 TI - Cardinal sutures revisited: position modification in corneal transplantation. PMID- 20010795 TI - The surgical outcome and personality change in a child with congenital cataract after multifocal intraocular lens implantation. PMID- 20010796 TI - Screening and characterization of microorganisms with FR901379 acylase activity. AB - A simple and rapid screen was performed for microorganisms producing cyclic lipopeptide acylase, an enzyme that deacylates the acyl side chain of FR901379 to yield a cyclic peptide, FR179642, which is an important intermediate in producing micafungin. Among approximately 3800 newly isolated strains from soil samples, 5 microorganisms expressing high acylase activity were selected and classified, 3 as Streptomyces spp. and 2 as filamentous fungi. This is the first time that these strains have been identified as cyclic lipopeptide acylase producers. Both fungi and streptomycetes showed differing optimum pH and temperature profiles for acylase activity. PMID- 20010797 TI - Constructing ribosomes along the Danube. AB - The EMBO Conference on Ribosome Synthesis held last summer explored the latest breakthroughs in ribosome assembly and how it affects disease. Both of these topics have recently seen important advances that enlighten how almost 200 proteins cooperate to produce a ribosome and how the cell responds to a malfunction in this process. PMID- 20010799 TI - The mousepox experience. An interview with Ronald Jackson and Ian Ramshaw on dual use research. Interview by Michael J. Selgelid and Lorna Weir. PMID- 20010798 TI - Coordination of genomic structure and transcription by the main bacterial nucleoid-associated protein HU. AB - The histone-like protein HU is a highly abundant DNA architectural protein that is involved in compacting the DNA of the bacterial nucleoid and in regulating the main DNA transactions, including gene transcription. However, the coordination of the genomic structure and function by HU is poorly understood. Here, we address this question by comparing transcript patterns and spatial distributions of RNA polymerase in Escherichia coli wild-type and hupA/B mutant cells. We demonstrate that, in mutant cells, upregulated genes are preferentially clustered in a large chromosomal domain comprising the ribosomal RNA operons organized on both sides of OriC. Furthermore, we show that, in parallel to this transcription asymmetry, mutant cells are also impaired in forming the transcription foci-spatially confined aggregations of RNA polymerase molecules transcribing strong ribosomal RNA operons. Our data thus implicate HU in coordinating the global genomic structure and function by regulating the spatial distribution of RNA polymerase in the nucleoid. PMID- 20010800 TI - Intrauterine programming of ageing. AB - Ageing is an unavoidable corollary to being alive; the most intuitive interpretation of ageing being that it is the consequence of progressive body degeneration. In agreement with this, current models propose that ageing occurs through a stepwise accumulation of DNA damage, which ultimately limits the regenerative capacity of tissues. On the other hand, there is increasing evidence that fetal distress can influence the development of disease in adult life, a phenomenon known as 'intrauterine programming'. The extent to which an intrauterine exposure to DNA damage can compromise lifespan remains unclear. My group has recently generated a murine model of a human syndrome linked to defective DNA repair and observed that these animals age prematurely, but the accumulation of DNA damage is restricted mostly to the embryonic period. Here, I discuss the implications of this finding and propose that ageing can be influenced by fetal distress. PMID- 20010801 TI - The candid approach. Scientists should insist on a purely scientific approach to preserving biodiversity. PMID- 20010802 TI - Nix is a selective autophagy receptor for mitochondrial clearance. AB - Autophagy is the cellular homeostatic pathway that delivers large cytosolic materials for degradation in the lysosome. Recent evidence indicates that autophagy mediates selective removal of protein aggregates, organelles and microbes in cells. Yet, the specificity in targeting a particular substrate to the autophagy pathway remains poorly understood. Here, we show that the mitochondrial protein Nix is a selective autophagy receptor by binding to LC3/GABARAP proteins, ubiquitin-like modifiers that are required for the growth of autophagosomal membranes. In cultured cells, Nix recruits GABARAP-L1 to damaged mitochondria through its amino-terminal LC3-interacting region. Furthermore, ablation of the Nix:LC3/GABARAP interaction retards mitochondrial clearance in maturing murine reticulocytes. Thus, Nix functions as an autophagy receptor, which mediates mitochondrial clearance after mitochondrial damage and during erythrocyte differentiation. PMID- 20010803 TI - RNA-directed DNA methylation and plant development require an IWR1-type transcription factor. AB - RNA-directed DNA methylation (RdDM) in plants requires two RNA polymerase (Pol) II-related RNA polymerases, namely Pol IV and Pol V. A genetic screen designed to reveal factors that are important for RdDM in a developmental context in Arabidopsis identified DEFECTIVE IN MERISTEM SILENCING 4 (DMS4). Unlike other mutants defective in RdDM, dms4 mutants have a pleiotropic developmental phenotype. The DMS4 protein is similar to yeast IWR1 (interacts with RNA polymerase II), a conserved putative transcription factor that interacts with Pol II subunits. The DMS4 complementary DNA partly complements the K1 killer toxin hypersensitivity of a yeast iwr1 mutant, suggesting some functional conservation. In the transgenic system studied, mutations in DMS4 directly or indirectly affect Pol IV-dependent secondary short interfering RNAs, Pol V-mediated RdDM, Pol V dependent synthesis of intergenic non-coding RNA and expression of many Pol II driven genes. These data suggest that DMS4 might be a regulatory factor for several RNA polymerases, thus explaining its diverse roles in the plant. PMID- 20010804 TI - Intergenic transcripts originating from a subclass of ribosomal DNA repeats silence ribosomal RNA genes in trans. AB - Epigenetic silencing of a fraction of ribosomal DNA (rDNA) requires association of the nucleolar chromatin-remodelling complex NoRC to 150-250 nucleotide RNAs (pRNA) that originate from an RNA polymerase I promoter located in the intergenic spacer separating rDNA repeats. Here, we show that NoRC-associated pRNA is transcribed from a sub-fraction of hypomethylated rRNA genes during mid S phase, acting in trans to inherit DNA methylation and transcriptional repression of late replicating silent rDNA copies. The results reveal variability between individual rDNA clusters with distinct functional consequences. PMID- 20010805 TI - The nuclear cofactor DOR regulates autophagy in mammalian and Drosophila cells. AB - The regulation of autophagy in metazoans is only partly understood, and there is a need to identify the proteins that control this process. The diabetes- and obesity-regulated gene (DOR), a recently reported nuclear cofactor of thyroid hormone receptors, is expressed abundantly in metabolically active tissues such as muscle. Here, we show that DOR shuttles between the nucleus and the cytoplasm, depending on cellular stress conditions, and re-localizes to autophagosomes on autophagy activation. We demonstrate that DOR interacts physically with autophagic proteins Golgi-associated ATPase enhancer of 16 kDa (GATE16) and microtubule-associated protein 1A/1B-light chain 3. Gain-of-function and loss-of function studies indicate that DOR stimulates autophagosome formation and accelerates the degradation of stable proteins. CG11347, the DOR Drosophila homologue, has been predicted to interact with the Drosophila Atg8 homologues, which suggests functional conservation in autophagy. Flies lacking CG11347 show reduced autophagy in the fat body during pupal development. All together, our data indicate that DOR regulates autophagosome formation and protein degradation in mammalian and Drosophila cells. PMID- 20010806 TI - DNA nanomechanics allows direct digital detection of complementary DNA and microRNA targets. AB - Techniques to detect and quantify DNA and RNA molecules in biological samples have had a central role in genomics research. Over the past decade, several techniques have been developed to improve detection performance and reduce the cost of genetic analysis. In particular, significant advances in label-free methods have been reported. Yet detection of DNA molecules at concentrations below the femtomolar level requires amplified detection schemes. Here we report a unique nanomechanical response of hybridized DNA and RNA molecules that serves as an intrinsic molecular label. Nanomechanical measurements on a microarray surface have sufficient background signal rejection to allow direct detection and counting of hybridized molecules. The digital response of the sensor provides a large dynamic range that is critical for gene expression profiling. We have measured differential expressions of microRNAs in tumour samples; such measurements have been shown to help discriminate between the tissue origins of metastatic tumours. Two hundred picograms of total RNA is found to be sufficient for this analysis. In addition, the limit of detection in pure samples is found to be one attomolar. These results suggest that nanomechanical read-out of microarrays promises attomolar-level sensitivity and large dynamic range for the analysis of gene expression, while eliminating biochemical manipulations, amplification and labelling. PMID- 20010807 TI - Secreted semaphorins control spine distribution and morphogenesis in the postnatal CNS. AB - The majority of excitatory synapses in the mammalian CNS (central nervous system) are formed on dendritic spines, and spine morphology and distribution are critical for synaptic transmission, synaptic integration and plasticity. Here, we show that a secreted semaphorin, Sema3F, is a negative regulator of spine development and synaptic structure. Mice with null mutations in genes encoding Sema3F, and its holoreceptor components neuropilin-2 (Npn-2, also known as Nrp2) and plexin A3 (PlexA3, also known as Plxna3), exhibit increased dentate gyrus (DG) granule cell (GC) and cortical layer V pyramidal neuron spine number and size, and also aberrant spine distribution. Moreover, Sema3F promotes loss of spines and excitatory synapses in dissociated neurons in vitro, and in Npn-2(-/-) brain slices cortical layer V and DG GCs exhibit increased mEPSC (miniature excitatory postsynaptic current) frequency. In contrast, a distinct Sema3A-Npn 1/PlexA4 signalling cascade controls basal dendritic arborization in layer V cortical neurons, but does not influence spine morphogenesis or distribution. These disparate effects of secreted semaphorins are reflected in the restricted dendritic localization of Npn-2 to apical dendrites and of Npn-1 (also known as Nrp1) to all dendrites of cortical pyramidal neurons. Therefore, Sema3F signalling controls spine distribution along select dendritic processes, and distinct secreted semaphorin signalling events orchestrate CNS connectivity through the differential control of spine morphogenesis, synapse formation, and the elaboration of dendritic morphology. PMID- 20010808 TI - HnRNP proteins controlled by c-Myc deregulate pyruvate kinase mRNA splicing in cancer. AB - When oxygen is abundant, quiescent cells efficiently extract energy from glucose primarily by oxidative phosphorylation, whereas under the same conditions tumour cells consume glucose more avidly, converting it to lactate. This long-observed phenomenon is known as aerobic glycolysis, and is important for cell growth. Because aerobic glycolysis is only useful to growing cells, it is tightly regulated in a proliferation-linked manner. In mammals, this is partly achieved through control of pyruvate kinase isoform expression. The embryonic pyruvate kinase isoform, PKM2, is almost universally re-expressed in cancer, and promotes aerobic glycolysis, whereas the adult isoform, PKM1, promotes oxidative phosphorylation. These two isoforms result from mutually exclusive alternative splicing of the PKM pre-mRNA, reflecting inclusion of either exon 9 (PKM1) or exon 10 (PKM2). Here we show that three heterogeneous nuclear ribonucleoprotein (hnRNP) proteins, polypyrimidine tract binding protein (PTB, also known as hnRNPI), hnRNPA1 and hnRNPA2, bind repressively to sequences flanking exon 9, resulting in exon 10 inclusion. We also demonstrate that the oncogenic transcription factor c-Myc upregulates transcription of PTB, hnRNPA1 and hnRNPA2, ensuring a high PKM2/PKM1 ratio. Establishing a relevance to cancer, we show that human gliomas overexpress c-Myc, PTB, hnRNPA1 and hnRNPA2 in a manner that correlates with PKM2 expression. Our results thus define a pathway that regulates an alternative splicing event required for tumour cell proliferation. PMID- 20010810 TI - Label-free, normalized quantification of complex mass spectrometry data for proteomic analysis. AB - Replicate mass spectrometry (MS) measurements and the use of multiple analytical methods can greatly expand the comprehensiveness of shotgun proteomic profiling of biological samples. However, the inherent biases and variations in such data create computational and statistical challenges for quantitative comparative analysis. We developed and tested a normalized, label-free quantitative method termed the normalized spectral index (SI(N)), which combines three MS abundance features: peptide count, spectral count and fragment-ion (tandem MS or MS/MS) intensity. SI(N) largely eliminated variances between replicate MS measurements, permitting quantitative reproducibility and highly significant quantification of thousands of proteins detected in replicate MS measurements of the same and distinct samples. It accurately predicts protein abundance more often than the five other methods we tested. Comparative immunoblotting and densitometry further validate our method. Comparative quantification of complex data sets from multiple shotgun proteomics measurements is relevant for systems biology and biomarker discovery. PMID- 20010809 TI - The sequence and de novo assembly of the giant panda genome. AB - Using next-generation sequencing technology alone, we have successfully generated and assembled a draft sequence of the giant panda genome. The assembled contigs (2.25 gigabases (Gb)) cover approximately 94% of the whole genome, and the remaining gaps (0.05 Gb) seem to contain carnivore-specific repeats and tandem repeats. Comparisons with the dog and human showed that the panda genome has a lower divergence rate. The assessment of panda genes potentially underlying some of its unique traits indicated that its bamboo diet might be more dependent on its gut microbiome than its own genetic composition. We also identified more than 2.7 million heterozygous single nucleotide polymorphisms in the diploid genome. Our data and analyses provide a foundation for promoting mammalian genetic research, and demonstrate the feasibility for using next-generation sequencing technologies for accurate, cost-effective and rapid de novo assembly of large eukaryotic genomes. PMID- 20010811 TI - Maintenance of a constitutive heterochromatin domain in vertebrates by a Dicer dependent mechanism. AB - The 16 kilobase (kb) heterochromatin domain between the chicken beta-globin locus and the folate receptor gene is used here to study the roles of RNA-dependent mechanisms and histone modifications in the maintenance of a constitutive heterochromatic structure. Inhibition of histone deacetylase (HDAC) activity is shown to both increase intergenic transcription and render the heterochromatin more accessible to MspI digestion. We show that short interfering RNA (siRNA) mediated downregulation of the enzyme Dicer has similar effects: histone acetylation is increased, transcript levels rise and the compact chromatin structure becomes more accessible to restriction endonucleases. We also show that the chicken Argonaute 2 homologue binds the 16 kb region in a Dicer-dependent manner and is necessary for a condensed chromatin structure. Heterochromatic domains of this kind, which are widely distributed in vertebrate genomes, thus seem to be maintained in their condensed form by highly conserved mechanisms. PMID- 20010812 TI - Carbonic anhydrases are upstream regulators of CO2-controlled stomatal movements in guard cells. AB - The continuing rise in atmospheric CO2 causes stomatal pores in leaves to close and thus globally affects CO2 influx into plants, water use efficiency and leaf heat stress. However, the CO2-binding proteins that control this response remain unknown. Moreover, which cell type responds to CO2, mesophyll or guard cells, and whether photosynthesis mediates this response are matters of debate. We demonstrate that Arabidopsis thaliana double-mutant plants in the beta-carbonic anhydrases betaCA1 and betaCA4 show impaired CO2-regulation of stomatal movements and increased stomatal density, but retain functional abscisic-acid and blue light responses. betaCA-mediated CO2-triggered stomatal movements are not, in first-order, linked to whole leaf photosynthesis and can function in guard cells. Furthermore, guard cell betaca-overexpressing plants exhibit instantaneous enhanced water use efficiency. Guard cell expression of mammalian alphaCAII complements the reduced sensitivity of ca1 ca4 plants, showing that carbonic anhydrase-mediated catalysis is an important mechanism for betaCA-mediated CO2 induced stomatal closure and patch clamp analyses indicate that CO2/HCO3- transfers the signal to anion channel regulation. These findings, together with ht1-2 (ref. 9) epistasis analysis demonstrate that carbonic anhydrases function early in the CO2 signalling pathway, which controls gas-exchange between plants and the atmosphere. PMID- 20010813 TI - Defects in DNA ligase I trigger PCNA ubiquitylation at Lys 107. AB - In all eukaryotes, the ligation of newly synthesized DNA, also known as Okazaki fragments, is catalysed by DNA ligase I (ref. 1). An individual with a DNA ligase I deficiency exhibits growth retardation, sunlight sensitivity and severe immunosuppression, probably due to accumulation of DNA damage. Surprisingly, not much is known about the DNA damage response (DDR) in DNA ligase I-deficient cells. As DNA replication and DDR pathways are highly conserved in eukaryotes, we used Saccharomyces cerevisiae as a model system to address this issue. We uncovered a new pathway, which facilitates ubiquitylation at Lys 107 of proliferating cell nuclear antigen (PCNA). Unlike ubiquitylation at Lys 164 of PCNA in response to UV irradiation, which triggers translesion synthesis, modification of Lys 107 is not dependent on the ubiquitin conjugating enzyme (E2) Rad6 (ref. 4) nor the ubiquitin ligase (E3) Rad18 (ref. 5), but requires the E2 variant Mms2 (ref. 6) in conjunction with Ubc4 (ref. 7) and the E3 Rad5 (Refs 8, 9). Surprisingly, DNA ligase I-deficient S. cerevisiae cdc9-1 cells that carry a PCNAK107R mutation are inviable, because they cannot activate a robust DDR. Furthermore, we show that ubiquitylation of PCNA in response to DNA ligase I deficiency is conserved in humans, yet the lysine residue that is modified remains to be determined. We propose that PCNA ubiquitylation provides a 'DNA damage code' that allows cells to categorize different types of defects that arise during DNA replication. PMID- 20010814 TI - A bacterial E3 ubiquitin ligase IpaH9.8 targets NEMO/IKKgamma to dampen the host NF-kappaB-mediated inflammatory response. AB - NF-kappaB (nuclear factor kappaB) has a pivotal role in many cellular processes, including the inflammatory and immune responses and, therefore, its activation is tightly regulated by the IKK (IkappaB kinase) complex and by IkappaBalpha degradation. When Shigella bacteria multiply within epithelial cells they release peptidoglycans, which are recognized by Nod1 and stimulate the NF-kappaB pathway, thus leading to a severe inflammatory response. Here, we show that IpaH9.8, a Shigella effector possessing E3 ligase activity, dampens the NF-kappaB-mediated inflammatory response to the bacterial infection in a unique way. IpaH9.8 interacts with NEMO/IKKgamma and ABIN-1, a ubiquitin-binding adaptor protein, promoting ABIN-1-dependent polyubiquitylation of NEMO. Consequently, polyubiquitylated NEMO undergoes proteasome-dependent degradation, which perturbs NF-kappaB activation. As NEMO is essential for NF-kappaB activation, we propose that the polyubiquitylation and degradation of NEMO during Shigella infection is a new bacterial strategy to modulate host inflammatory responses. PMID- 20010815 TI - Cdk2 suppresses cellular senescence induced by the c-myc oncogene. AB - Activated oncogenes induce compensatory tumour-suppressive responses, such as cellular senescence or apoptosis, but the signals determining the main outcome remain to be fully understood. Here, we uncover a role for Cdk2 (cyclin-dependent kinase 2) in suppressing Myc-induced senescence. Short-term activation of Myc promoted cell-cycle progression in either wild-type or Cdk2 knockout mouse embryo fibroblasts (MEFs). In the knockout MEFs, however, the initial hyper proliferative response was followed by cellular senescence. Loss of Cdk2 also caused sensitization to Myc-induced senescence in pancreatic beta-cells or splenic B-cells in vivo, correlating with delayed lymphoma onset in the latter. Cdk2-/- MEFs also senesced upon ectopic Wnt signalling or, without an oncogene, upon oxygen-induced culture shock. Myc also causes senescence in cells lacking the DNA repair protein Wrn. However, unlike loss of Wrn, loss of Cdk2 did not enhance Myc-induced replication stress, implying that these proteins suppress senescence through different routes. In MEFs, Myc-induced senescence was genetically dependent on the ARF-p53-p21Cip1 and p16INK4a-pRb pathways, p21Cip1 and p16INK4a being selectively induced in Cdk2-/- cells. Thus, although redundant for cell-cycle progression and development, Cdk2 has a unique role in suppressing oncogene- and/or stress-induced senescence. Pharmacological inhibition of Cdk2 induced Myc-dependent senescence in various cell types, including a p53-null human cancer cell line. Our data warrant re-assessment of Cdk2 as a therapeutic target in Myc- or Wnt-driven tumours. PMID- 20010816 TI - A role for Rho GTPases and cell-cell adhesion in single-cell motility in vivo. AB - Cell migration is central to embryonic development, homeostasis and disease, processes in which cells move as part of a group or individually. Whereas the mechanisms controlling single-cell migration in vitro are relatively well understood, less is known about the mechanisms promoting the motility of individual cells in vivo. In particular, it is not clear how cells that form blebs in their migration use those protrusions to bring about movement in the context of the three-dimensional cellular environment. Here we show that the motility of chemokine-guided germ cells within the zebrafish embryo requires the function of the small Rho GTPases Rac1 and RhoA, as well as E-cadherin-mediated cell-cell adhesion. Using fluorescence resonance energy transfer we demonstrate that Rac1 and RhoA are activated in the cell front. At this location, Rac1 is responsible for the formation of actin-rich structures, and RhoA promotes retrograde actin flow. We propose that these actin-rich structures undergoing retrograde flow are essential for the generation of E-cadherin-mediated traction forces between the germ cells and the surrounding tissue and are therefore crucial for cell motility in vivo. PMID- 20010817 TI - Orphan nuclear receptor TLX activates Wnt/beta-catenin signalling to stimulate neural stem cell proliferation and self-renewal. AB - The nuclear receptor TLX (also known as NR2E1) is essential for adult neural stem cell self-renewal; however, the molecular mechanisms involved remain elusive. Here we show that TLX activates the canonical Wnt/beta-catenin pathway in adult mouse neural stem cells. Furthermore, we demonstrate that Wnt/beta-catenin signalling is important in the proliferation and self-renewal of adult neural stem cells in the presence of epidermal growth factor and fibroblast growth factor. Wnt7a and active beta-catenin promote neural stem cell self-renewal, whereas the deletion of Wnt7a or the lentiviral transduction of axin, a beta catenin inhibitor, led to decreased cell proliferation in adult neurogenic areas. Lentiviral transduction of active beta-catenin led to increased numbers of type B neural stem cells in the subventricular zone of adult brains, whereas deletion of Wnt7a or TLX resulted in decreased numbers of neural stem cells retaining bromodeoxyuridine label in the adult brain. Both Wnt7a and active beta-catenin significantly rescued a TLX (also known as Nr2e1) short interfering RNA-induced deficiency in neural stem cell proliferation. Lentiviral transduction of an active beta-catenin increased cell proliferation in neurogenic areas of TLX-null adult brains markedly. These results strongly support the hypothesis that TLX acts through the Wnt/beta-catenin pathway to regulate neural stem cell proliferation and self-renewal. Moreover, this study suggests that neural stem cells can promote their own self-renewal by secreting signalling molecules that act in an autocrine/paracrine mode. PMID- 20010818 TI - An in vivo biosensor for neurotransmitter release and in situ receptor activity. AB - Tools from molecular biology, combined with in vivo optical imaging techniques, provide new mechanisms for noninvasively observing brain processes. Current approaches primarily probe cell-based variables, such as cytosolic calcium or membrane potential, but not cell-to-cell signaling. We devised cell-based neurotransmitter fluorescent engineered reporters (CNiFERs) to address this challenge and monitor in situ neurotransmitter receptor activation. CNiFERs are cultured cells that are engineered to express a chosen metabotropic receptor, use the G(q) protein-coupled receptor cascade to transform receptor activity into a rise in cytosolic [Ca(2+)] and report [Ca(2+)] with a genetically encoded fluorescent Ca(2+) sensor. The initial realization of CNiFERs detected acetylcholine release via activation of M1 muscarinic receptors. We used chronic implantation of M1-CNiFERs in frontal cortex of the adult rat to elucidate the muscarinic action of the atypical neuroleptics clozapine and olanzapine. We found that these drugs potently inhibited in situ muscarinic receptor activity. PMID- 20010819 TI - PIP3 controls synaptic function by maintaining AMPA receptor clustering at the postsynaptic membrane. AB - Despite their low abundance, phosphoinositides are critical regulators of intracellular signaling and membrane compartmentalization. However, little is known of phosphoinositide function at the postsynaptic membrane. Here we show that continuous synthesis and availability of phosphatidylinositol-(3,4,5) trisphosphate (PIP(3)) at the postsynaptic terminal is necessary for sustaining synaptic function in rat hippocampal neurons. This requirement was specific for synaptic, but not extrasynaptic, AMPA receptors, nor for NMDA receptors. PIP(3) downregulation impaired PSD-95 accumulation in spines. Concomitantly, AMPA receptors became more mobile and migrated from the postsynaptic density toward the perisynaptic membrane within the spine, leading to synaptic depression. Notably, these effects were only revealed after prolonged inhibition of PIP(3) synthesis or by direct quenching of this phosphoinositide at the postsynaptic cell. Therefore, we conclude that a slow, but constant, turnover of PIP(3) at synapses is required for maintaining AMPA receptor clustering and synaptic strength under basal conditions. PMID- 20010820 TI - A small number of open Ca2+ channels trigger transmitter release at a central GABAergic synapse. AB - To determine the number of open Ca(2+) channels necessary for transmitter release at the inhibitory basket cell-granule cell synapse in rat hippocampus, we combined presynaptic Ca(2+) imaging, recording of postsynaptic currents and modeling. We found that that the opening of three or fewer Ca(2+) channels triggered transmitter release. Furthermore, a small number of Ca(2+) channels were able to evoke release with high temporal precision, despite stochastic Ca(2+) channel opening. PMID- 20010821 TI - Synaptotagmin IV determines the linear Ca2+ dependence of vesicle fusion at auditory ribbon synapses. AB - Mammalian cochlear inner hair cells (IHCs) are specialized for the dynamic coding of continuous and finely graded sound signals. This ability is largely conferred by the linear Ca(2+) dependence of neurotransmitter release at their synapses, which is also a feature of visual and olfactory systems. The prevailing hypothesis is that linearity in IHCs occurs through a developmental change in the Ca(2+) sensitivity of synaptic vesicle fusion from the nonlinear (high order) Ca(2+) dependence of immature spiking cells. However, the nature of the Ca(2+) sensor(s) of vesicle fusion at hair cell synapses is unknown. We found that synaptotagmin IV was essential for establishing the linear exocytotic Ca(2+) dependence in adult rodent IHCs and immature outer hair cells. Moreover, the expression of the hitherto undetected synaptotagmins I and II correlated with a high-order Ca(2+) dependence in IHCs. We propose that the differential expression of synaptotagmins determines the characteristic Ca(2+) sensitivity of vesicle fusion at hair cell synapses. PMID- 20010822 TI - SLEEPLESS, a Ly-6/neurotoxin family member, regulates the levels, localization and activity of Shaker. AB - Sleep is a whole-organism phenomenon accompanied by global changes in neural activity. We previously identified SLEEPLESS (SSS) as a glycosylphosphatidyl inositol-anchored protein required for sleep in Drosophila. Here we found that SSS is critical for regulating the sleep-modulating potassium channel Shaker. SSS and Shaker shared similar expression patterns in the brain and specifically affected each other's expression levels. sleepless (sss) loss-of-function mutants exhibited altered Shaker localization, reduced Shaker current density and slower Shaker current kinetics. Transgenic expression of sss in sss mutants rescued defects in Shaker expression and activity cell-autonomously and suggested that SSS functions in wake-promoting, cholinergic neurons. In heterologous cells, SSS accelerated the kinetics of Shaker currents and was co-immunoprecipitated with Shaker, suggesting that SSS modulates Shaker activity via a direct interaction. SSS is predicted to belong to the Ly-6/neurotoxin superfamily, suggesting a mechanism for regulation of neuronal excitability by endogenous toxin-like molecules. PMID- 20010823 TI - Synaptic computation underlying probabilistic inference. AB - We propose that synapses may be the workhorse of the neuronal computations that underlie probabilistic reasoning. We built a neural circuit model for probabilistic inference in which information provided by different sensory cues must be integrated and the predictive powers of individual cues about an outcome are deduced through experience. We found that bounded synapses naturally compute, through reward-dependent plasticity, the posterior probability that a choice alternative is correct given that a cue is presented. Furthermore, a decision circuit endowed with such synapses makes choices on the basis of the summed log posterior odds and performs near-optimal cue combination. The model was validated by reproducing salient observations of, and provides insights into, a monkey experiment using a categorization task. Our model thus suggests a biophysical instantiation of the Bayesian decision rule, while predicting important deviations from it similar to the 'base-rate neglect' observed in human studies when alternatives have unequal prior probabilities. PMID- 20010824 TI - Rewiring of hindlimb corticospinal neurons after spinal cord injury. AB - Little is known about the functional role of axotomized cortical neurons that survive spinal cord injury. Large thoracic spinal cord injuries in adult rats result in impairments of hindlimb function. Using retrograde tracers, we found that axotomized corticospinal axons from the hindlimb sensorimotor cortex sprouted in the cervical spinal cord. Mapping of these neurons revealed the emergence of a new forelimb corticospinal projection from the rostral part of the former hindlimb cortex. Voltage-sensitive dye (VSD) imaging and blood-oxygen level-dependent functional magnetic resonance imaging (BOLD fMRI) revealed a stable expansion of the forelimb sensory map, covering in particular the former hindlimb cortex containing the rewired neurons. Therefore, axotomized hindlimb corticospinal neurons can be incorporated into the sensorimotor circuits of the unaffected forelimb. PMID- 20010825 TI - Label-free biomarker detection from whole blood. AB - Label-free nanosensors can detect disease markers to provide point-of-care diagnosis that is low-cost, rapid, specific and sensitive. However, detecting these biomarkers in physiological fluid samples is difficult because of problems such as biofouling and non-specific binding, and the resulting need to use purified buffers greatly reduces the clinical relevance of these sensors. Here, we overcome this limitation by using distinct components within the sensor to perform purification and detection. A microfluidic purification chip simultaneously captures multiple biomarkers from blood samples and releases them, after washing, into purified buffer for sensing by a silicon nanoribbon detector. This two-stage approach isolates the detector from the complex environment of whole blood, and reduces its minimum required sensitivity by effectively pre concentrating the biomarkers. We show specific and quantitative detection of two model cancer antigens from a 10 microl sample of whole blood in less than 20 min. This study marks the first use of label-free nanosensors with physiological solutions, positioning this technology for rapid translation to clinical settings. PMID- 20010826 TI - Aharonov-Bohm interference in topological insulator nanoribbons. AB - Topological insulators represent unusual phases of quantum matter with an insulating bulk gap and gapless edges or surface states. The two-dimensional topological insulator phase was predicted in HgTe quantum wells and confirmed by transport measurements. Recently, Bi(2)Se(3) and related materials have been proposed as three-dimensional topological insulators with a single Dirac cone on the surface, protected by time-reversal symmetry. The topological surface states have been observed by angle-resolved photoemission spectroscopy experiments. However, few transport measurements in this context have been reported, presumably owing to the predominance of bulk carriers from crystal defects or thermal excitations. Here we show unambiguous transport evidence of topological surface states through periodic quantum interference effects in layered single crystalline Bi(2)Se(3) nanoribbons, which have larger surface-to-volume ratios than bulk materials and can therefore manifest surface effects. Pronounced Aharonov-Bohm oscillations in the magnetoresistance clearly demonstrate the coherent propagation of two-dimensional electrons around the perimeter of the nanoribbon surface, as expected from the topological nature of the surface states. The dominance of the primary h/e oscillation, where h is Planck's constant and e is the electron charge, and its temperature dependence demonstrate the robustness of these states. Our results suggest that topological insulator nanoribbons afford promising materials for future spintronic devices at room temperature. PMID- 20010827 TI - Porous metal-organic-framework nanoscale carriers as a potential platform for drug delivery and imaging. AB - In the domain of health, one important challenge is the efficient delivery of drugs in the body using non-toxic nanocarriers. Most of the existing carrier materials show poor drug loading (usually less than 5 wt% of the transported drug versus the carrier material) and/or rapid release of the proportion of the drug that is simply adsorbed (or anchored) at the external surface of the nanocarrier. In this context, porous hybrid solids, with the ability to tune their structures and porosities for better drug interactions and high loadings, are well suited to serve as nanocarriers for delivery and imaging applications. Here we show that specific non-toxic porous iron(III)-based metal-organic frameworks with engineered cores and surfaces, as well as imaging properties, function as superior nanocarriers for efficient controlled delivery of challenging antitumoural and retroviral drugs (that is, busulfan, azidothymidine triphosphate, doxorubicin or cidofovir) against cancer and AIDS. In addition to their high loadings, they also potentially associate therapeutics and diagnostics, thus opening the way for theranostics, or personalized patient treatments. PMID- 20010828 TI - Oscillatory spin-polarized tunnelling from silicon quantum wells controlled by electric field. AB - Spin-dependent electronic transport is widely used to probe and manipulate magnetic materials and develop spin-based devices. Spin-polarized tunnelling, successful in ferromagnetic metal junctions, was recently used to inject and detect electron spins in organics and bulk GaAs or Si. Electric field control of spin precession was studied in III-V semiconductors relying on spin-orbit interaction, which makes this approach inefficient for Si, the mainstream semiconductor. Methods to control spin other than through precession are thus desired. Here we demonstrate electrostatic modification of the magnitude of spin polarization in a silicon quantum well, and detection thereof by means of tunnelling to a ferromagnet, producing prominent oscillations of tunnel magnetoresistance of up to 8%. The electric modification of the spin polarization relies on discrete states in the Si with a Zeeman spin splitting, an approach that is also applicable to organic, carbon-based and other materials with weak spin-orbit interaction. PMID- 20010829 TI - Nanogap-engineerable Raman-active nanodumbbells for single-molecule detection. AB - Surface-enhanced Raman scattering (SERS)-based signal amplification and detection methods using plasmonic nanostructures have been widely investigated for imaging and sensing applications. However, SERS-based molecule detection strategies have not been practically useful because there is no straightforward method to synthesize and characterize highly sensitive SERS-active nanostructures with sufficiently high yield and efficiency, which results in an extremely low cross section area in Raman sensing. Here, we report a high-yield synthetic method for SERS-active gold-silver core-shell nanodumbbells, where the gap between two nanoparticles and the Raman-dye position and environment can be engineered on the nanoscale. Atomic-force-microscope-correlated nano-Raman measurements of individual dumbbell structures demonstrate that Raman signals can be repeatedly detected from single-DNA-tethered nanodumbbells. These programmed nanostructure fabrication and single-DNA detection strategies open avenues for the high-yield synthesis of optically active smart nanoparticles and structurally reproducible nanostructure-based single-molecule detection and bioassays. PMID- 20010830 TI - Explaining the paradoxical diversity of ultrafast laser-induced demagnetization. AB - Pulsed-laser-induced quenching of ferromagnetic order has intrigued researchers since pioneering works in the 1990s. It was reported that demagnetization in gadolinium proceeds within 100 ps, but three orders of magnitude faster in ferromagnetic transition metals such as nickel. Here we show that a model based on electron-phonon-mediated spin-flip scattering explains both timescales on equal footing. Our interpretation is supported by ab initio estimates of the spin flip scattering probability, and experimental fluence dependencies are shown to agree perfectly with predictions. A phase diagram is constructed in which two classes of laser-induced magnetization dynamics can be distinguished, where the ratio of the Curie temperature to the atomic magnetic moment turns out to have a crucial role. We conclude that the ultrafast magnetization dynamics can be well described disregarding highly excited electronic states, merely considering the thermalized electron system. PMID- 20010831 TI - Single-gene transgenic mouse strains for reprogramming adult somatic cells. AB - We report transgenic mouse models in which three or four reprogramming factors are expressed from a single genomic locus using a drug-inducible transgene. Multiple somatic cell types can be directly reprogrammed to generate induced pluripotent stem cells (iPSCs) by culture in doxycycline. Because reprogramming factors are carried on a single polycistronic construct, the mice can be easily maintained, and the transgene can be easily transferred into other genetic backgrounds. PMID- 20010832 TI - A reprogrammable mouse strain from gene-targeted embryonic stem cells. AB - The derivation of induced pluripotent stem cells (iPSCs) usually involves the viral introduction of reprogramming factors into somatic cells. Here we used gene targeting to generate a mouse strain with a single copy of an inducible, polycistronic reprogramming cassette, allowing for the induction of pluripotency in various somatic cell types. As these 'reprogrammable mice' can be easily bred, they are a useful tool to study the mechanisms underlying cellular reprogramming. PMID- 20010833 TI - Integrated analysis of receptor activation and downstream signaling with EXTassays. AB - The ability to measure multiple cellular signaling events is essential to better understand the underlying complex biological processes that occur in living cells. Microarray-based technologies are now commonly used to study changes in transcription. This information, however, is not sufficient to understand the regulatory mechanisms that lead to gene expression changes. Here we present an approach to monitor signaling events upstream of gene expression. We coupled different reporter gene assays to unique expressed oligonucleotide tags (EXTs) that serve as identifiers and quantitative reporters. Multiple EXT reporters can be isolated as a pool and analyzed by hybridization to microarrays. To test the feasibility of our approach, we integrated complementation assays based on a protease from tobacco etch virus (TEV protease) and transcription factor activity profiling. Thereby, we simultaneously monitored Neuregulin-dependent mouse ErbB receptor tyrosine kinase dimerization, effector recruitment and downstream signaling. PMID- 20010834 TI - Genome-wide association study identifies five loci associated with lung function. AB - Pulmonary function measures are heritable traits that predict morbidity and mortality and define chronic obstructive pulmonary disease (COPD). We tested genome-wide association with forced expiratory volume in 1 s (FEV(1)) and the ratio of FEV(1) to forced vital capacity (FVC) in the SpiroMeta consortium (n = 20,288 individuals of European ancestry). We conducted a meta-analysis of top signals with data from direct genotyping (n < or = 32,184 additional individuals) and in silico summary association data from the CHARGE Consortium (n = 21,209) and the Health 2000 survey (n < or = 883). We confirmed the reported locus at 4q31 and identified associations with FEV(1) or FEV(1)/FVC and common variants at five additional loci: 2q35 in TNS1 (P = 1.11 x 10(-12)), 4q24 in GSTCD (2.18 x 10(-23)), 5q33 in HTR4 (P = 4.29 x 10(-9)), 6p21 in AGER (P = 3.07 x 10(-15)) and 15q23 in THSD4 (P = 7.24 x 10(-15)). mRNA analyses showed expression of TNS1, GSTCD, AGER, HTR4 and THSD4 in human lung tissue. These associations offer mechanistic insight into pulmonary function regulation and indicate potential targets for interventions to alleviate respiratory disease. PMID- 20010836 TI - Preferential associations between co-regulated genes reveal a transcriptional interactome in erythroid cells. AB - The discovery of interchromosomal interactions in higher eukaryotes points to a functional interplay between genome architecture and gene expression, challenging the view of transcription as a one-dimensional process. However, the extent of interchromosomal interactions and the underlying mechanisms are unknown. Here we present the first genome-wide analysis of transcriptional interactions using the mouse globin genes in erythroid tissues. Our results show that the active globin genes associate with hundreds of other transcribed genes, revealing extensive and preferential intra- and interchromosomal transcription interactomes. We show that the transcription factor Klf1 mediates preferential co-associations of Klf1 regulated genes at a limited number of specialized transcription factories. Our results establish a new gene expression paradigm, implying that active co regulated genes and their regulatory factors cooperate to create specialized nuclear hot spots optimized for efficient and coordinated transcriptional control. PMID- 20010835 TI - Meta-analyses of genome-wide association studies identify multiple loci associated with pulmonary function. AB - Spirometric measures of lung function are heritable traits that reflect respiratory health and predict morbidity and mortality. We meta-analyzed genome wide association studies for two clinically important lung-function measures: forced expiratory volume in the first second (FEV(1)) and its ratio to forced vital capacity (FEV(1)/FVC), an indicator of airflow obstruction. This meta analysis included 20,890 participants of European ancestry from four CHARGE Consortium studies: Atherosclerosis Risk in Communities, Cardiovascular Health Study, Framingham Heart Study and Rotterdam Study. We identified eight loci associated with FEV(1)/FVC (HHIP, GPR126, ADAM19, AGER-PPT2, FAM13A, PTCH1, PID1 and HTR4) and one locus associated with FEV(1) (INTS12-GSTCD-NPNT) at or near genome-wide significance (P < 5 x 10(-8)) in the CHARGE Consortium dataset. Our findings may offer insights into pulmonary function and pathogenesis of chronic lung disease. PMID- 20010837 TI - THAP proteins target specific DNA sites through bipartite recognition of adjacent major and minor grooves. AB - THAP-family C(2)CH zinc-coordinating DNA-binding proteins function in diverse eukaryotic cellular processes, such as transposition, transcriptional repression, stem-cell pluripotency, angiogenesis and neurological function. To determine the molecular basis for sequence-specific DNA recognition by THAP proteins, we solved the crystal structure of the Drosophila melanogaster P element transposase THAP domain (DmTHAP) in complex with a natural 10-base-pair site. In contrast to C(2)H(2) zinc fingers, DmTHAP docks a conserved beta-sheet into the major groove and a basic C-terminal loop into the adjacent minor groove. We confirmed specific protein-DNA interactions by mutagenesis and DNA-binding assays. Sequence analysis of natural and in vitro-selected binding sites suggests that several THAPs (DmTHAP and human THAP1 and THAP9) recognize a bipartite TXXGGGX(A/T) consensus motif; homology suggests THAP proteins bind DNA through a bipartite interaction. These findings reveal the conserved mechanisms by which THAP-family proteins engage specific chromosomal target elements. PMID- 20010838 TI - Structure and mechanism of a pentameric formate channel. AB - Formate transport across the inner membrane is a critical step in anaerobic bacterial respiration. Members of the formate/nitrite transport protein family function to shuttle substrate across the cytoplasmic membrane. In bacterial pathogens, the nitrite transport protein is involved in protecting bacteria from peroxynitrite released by host macrophages. We have determined the 2.13-A structure of the formate channel FocA from Vibrio cholerae, which reveals a pentamer in which each monomer possesses its own substrate translocation pore. Unexpectedly, the fold of the FocA monomer resembles that found in water and glycerol channels. The selectivity filter in FocA consists of a cytoplasmic slit and a central constriction ring. A 2.5-A high-formate structure shows two formate ions bound to the cytoplasmic slit via both hydrogen bonding and van der Waals interactions, providing a structural basis for the substrate selectivity of the channel. PMID- 20010839 TI - Modulation of protein properties in living cells using nanobodies. AB - Protein conformation is critically linked to function and often controlled by interactions with regulatory factors. Here we report the selection of camelid derived single-domain antibodies (nanobodies) that modulate the conformation and spectral properties of the green fluorescent protein (GFP). One nanobody could reversibly reduce GFP fluorescence by a factor of 5, whereas its displacement by a second nanobody caused an increase by a factor of 10. Structural analysis of GFP-nanobody complexes revealed that the two nanobodies induce subtle opposing changes in the chromophore environment, leading to altered absorption properties. Unlike conventional antibodies, the small, stable nanobodies are functional in living cells. Nanobody-induced changes were detected by ratio imaging and used to monitor protein expression and subcellular localization as well as translocation events such as the tamoxifen-induced nuclear localization of estrogen receptor. This work demonstrates that protein conformations can be manipulated and studied with nanobodies in living cells. PMID- 20010840 TI - Structure of the measles virus hemagglutinin bound to the CD46 receptor. AB - The highly contagious measles virus infects millions of individuals worldwide, causing serious disease in children of developing countries. Infection is initiated by attachment of the measles virus hemagglutinin (MV-H), a glycoprotein anchored to the virus envelope, to the host cell receptors CD46 or signaling lymphocyte activation molecule (SLAM). Here we report the crystal structure of MV H in complex with a CD46 protein spanning the two N-terminal domains. A unique groove at the side of the MV-H beta-propeller domain, which is absent in homologous paramyxovirus attachment proteins, engages residues in both CD46 domains. Key contacts involve a protruding loop in the N-terminal CD46 domain that carries two sequential proline residues (PP motif) and penetrates deeply into a hydrophobic socket in MV-H. We identify a similar PP motif in SLAM, defining a common measles virus recognition epitope in the CD46 and SLAM receptor proteins. PMID- 20010841 TI - A conserved activation element in BMP signaling during Drosophila development. AB - The transforming growth factor beta (TGF-beta) family member Decapentaplegic (Dpp) is a key regulator of patterning and growth in Drosophila development. Previous studies have identified a short DNA motif called the silencer element (SE), which recruits a trimeric Smad complex and the repressor Schnurri to downregulate target enhancers upon Dpp signaling. We have now isolated the minimal enhancer of the dad gene and discovered a short motif we termed the activating element (AE). The AE is similar to the SE and recruits the Smad proteins via a conserved mechanism. However, the AE and SE differ at important nucleotide positions. As a consequence, the AE does not recruit Schnurri but rather integrates repressive input by the default repressor Brinker and activating input by the Smad signal transducers Mothers against Dpp (Mad) and Medea via competitive DNA binding. The AE allows the identification of hitherto unknown direct Dpp targets and is functionally conserved in vertebrates. PMID- 20010842 TI - Structure of the MLL CXXC domain-DNA complex and its functional role in MLL-AF9 leukemia. AB - The gene MLL (encoding the protein mixed-lineage leukemia) is the target of chromosomal translocations that cause leukemias with poor prognosis. All leukemogenic MLL fusion proteins retain the CXXC domain, which binds to nonmethylated CpG DNA sites. We present the solution structure of the MLL CXXC domain in complex with DNA, showing how the CXXC domain distinguishes nonmethylated from methylated CpG DNA. On the basis of the structure, we generated point mutations that disrupt DNA binding. Introduction of these mutations into the MLL-AF9 fusion protein resulted in increased DNA methylation of specific CpG nucleotides in Hoxa9, increased H3K9 methylation, decreased expression of Hoxa9-locus transcripts, loss of immortalization potential, and inability to induce leukemia in mice. These results establish that DNA binding by the CXXC domain and protection against DNA methylation is essential for MLL fusion leukemia. They also provide support for viewing this interaction as a potential target for therapeutic intervention. PMID- 20010843 TI - Orthogonal use of a human tRNA synthetase active site to achieve multifunctionality. AB - Protein multifunctionality is an emerging explanation for the complexity of higher organisms. In this regard, aminoacyl tRNA synthetases catalyze amino acid activation for protein synthesis, but some also act in pathways for inflammation, angiogenesis and apoptosis. It is unclear how these multiple functions evolved and how they relate to the active site. Here structural modeling analysis, mutagenesis and cell-based functional studies show that the potent angiostatic, natural fragment of human tryptophanyl-tRNA synthetase (TrpRS) associates via tryptophan side chains that protrude from its cognate cellular receptor vascular endothelial cadherin (VE-cadherin). VE-cadherin's tryptophan side chains fit into the tryptophan-specific active site of the synthetase. Thus, specific side chains of the receptor mimic amino acid substrates and expand the functionality of the active site of the synthetase. We propose that orthogonal use of the same active site may be a general way to develop multifunctionality of human tRNA synthetases and other proteins. PMID- 20010844 TI - TCR and Lat are expressed on separate protein islands on T cell membranes and concatenate during activation. AB - The organization and dynamics of receptors and other molecules in the plasma membrane are not well understood. Here we analyzed the spatio-temporal dynamics of T cell antigen receptor (TCR) complexes and linker for activation of T cells (Lat), a key adaptor molecule in the TCR signaling pathway, in T cell membranes using high-speed photoactivated localization microscopy, dual-color fluorescence cross-correlation spectroscopy and transmission electron microscopy. In quiescent T cells, both molecules existed in separate membrane domains (protein islands), and these domains concatenated after T cell activation. These concatemers were identical to signaling microclusters, a prominent hallmark of T cell activation. This separation versus physical juxtapositioning of receptor domains and domains containing downstream signaling molecules in quiescent versus activated T cells may be a general feature of plasma membrane-associated signal transduction. PMID- 20010845 TI - CXCR4 acts as a costimulator during thymic beta-selection. AB - Passage through the beta-selection developmental checkpoint requires productive rearrangement of segments of the T cell antigen receptor-beta gene (Tcrb) and formation of a pre-TCR on the surface of CD4(-)CD8(-) thymocytes. How other receptors influence betabeta-selection is less well understood. Here we define a new role for the chemokine receptor CXCR4 during T cell development. CXCR4 functionally associated with the pre-TCR and influenced beta-selection by regulating the steady-state localization of immature thymocytes in thymic subregions, by facilitating optimal pre-TCR-induced survival signals, and by promoting thymocyte proliferation. We also characterize functionally relevant signaling molecules downstream of CXCR4 and the pre-TCR in thymocytes. Our data designate CXCR4 as a costimulator of the pre-TCR during beta-selection. PMID- 20010846 TI - Comparative study of the effects of three semen preparation media on semen analysis, DNA damage and protamine deficiency, and the correlation between DNA integrity and sperm parameters. AB - Semen samples collected from 28 male partners of infertile couples were divided into three equal aliquots and prepared with three selected media, such as PureSperm (Nidacon, Gothenburg, Sweden), Sil-Select Plus (Fertipro, Beernem, Belgium) and SpermGrad (Vitrolife, Gothenburg, Sweden). The differences in mean percentages of semen parameters were assessed by repeated measures analysis. Correlations of sperm DNA damage, as measured by terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay, and of protamine deficiency, as measured by chromomycin A3 (CMA3) staining with sperm parameters, were determined by Pearson's correlation. After preparation with all three media, sperm concentrations decreased (P < 0.05) while percentages of sperm with normal morphology increased (P < 0.05). Percentages of sperm motility, rapid motility and progressive motile concentration (PMC) increased (P < 0.05) for each of these parameters, PureSperm preparation gave the best results (P < 0.05). The percentage of DNA damage decreased in the PureSperm and Sil-Select Plus preparations (17.9% and 31.3%, respectively, P < 0.05) and increased in the SpermGrad preparation (56.3%, P < 0.05). Protamine deficiency also decreased in all three kinds of media, 59.3%, 47.7% and 40.3% for PureSperm, Sil-Select Plus and SpermGrad preparations, respectively (P < 0.05). The percentage of DNA damaged sperm was negatively correlated with the percentages of sperm motility, rapid motility and PMC, but was positively correlated with static motility (P < 0.05). This comparative study and correlation analysis revealed that PureSperm preparation yielded sperm with the best motility and the lowest percentage of protamine deficiency. The Sil-Select Plus preparation yielded sperm with the lowest amount of DNA damage. The SpermGrad preparation had a high percentage of sperm with normal morphology, but also had the highest percentage of sperm with DNA damage. Sperm DNA damage was correlated with percentages of sperm motility, rapid motility, static motility and PMC. PMID- 20010847 TI - Optimal dose of busulfan for depleting testicular germ cells of recipient mice before spermatogonial transplantation. AB - Successful spermatogonial transplantation requires depletion of the host germ cells to allow efficient colonization of the donor spermatogonial stem cells. Although a sterilizing drug, busulfan (Myleran), is commonly used for preparing a recipient mouse before transplantation, the optimal dose of this drug has not yet been defined. The present study investigated the effects of different doses of busulfan (10-50 mg per kg body weight) on survival rate, testicular mass and histomorphology, and on the haploid spermatids and spermatozoa of male BALB/c mice. The results suggest that a dosage of 30 mg kg(-1) is optimal for the ablative treatment with busulfan used to prepare the recipient mice. This dose results in an adequate depletion of the host germ cells for colonization of donor derived spermatogonial stem cells and causes the lowest death rate of the animals. PMID- 20010848 TI - The protective effects of alpha-ketoacids against oxidative stress on rat spermatozoa in vitro. AB - The aim of this study was to determine the effects of antioxidants, including alpha-ketoacids (alpha-ketoglutarate and pyruvate), lactate and glutamate/malate combination, against oxidative stress on rat spermatozoa. Our results showed that H(2)O(2) (250 micromol L(-1))-induced damages, such as impaired motility, adenosine triphosphate (ATP) depletion, inhibition of sperm protein phosphorylation, reduced acrosome reaction and decreased viability, could be significantly prevented by incubation of the spermatozoa with alpha-ketoglutarate (4 mmol L(-1)) or pyruvate (4 mmol L(-1)). Without exogenous H(2)O(2) in the medium, the addition of pyruvate (4 mmol L(-1)) significantly increased the superoxide anion (O(2)(-).) level in sperm suspension (P < or = 0.01), whereas the addition of alpha-ketoglutarate (4 mmol L(-1)) and lactate (4 mmol L(-1)) significantly enhanced tyrosine-phosphorylated proteins with the size of 95 kDa (P < or = 0.04). At the same time, alpha-ketoglutarate, pyruvate, lactate, glutamate and malate supplemented in media can be used as important energy sources and supply ATP for sperm motility. In conclusion, the present results show that alpha-ketoacids could be effective antioxidants for protecting rat spermatozoa from H(2)O(2) attack and could be effective components to improve the antioxidant capacity of Biggers, Whitten and Whittingham media. PMID- 20010849 TI - High-dose dietary zinc promotes prostate intraepithelial neoplasia in a murine tumor induction model. AB - To evaluate the role of high-dose dietary zinc in the process of prostate malignancy, 60 Sprague-Dawley rats were randomly divided into four groups: tumor induction with carcinogen and hormone (group 1), oral zinc administration without tumor induction (group 2), oral zinc administration with tumor induction (group 3) and a control without zinc administration or tumor induction (group 4). Zinc was supplied orally in the form of zinc sulfate heptahydrate dissolved in drinking water to groups 2 and 3 for 20 weeks. Although the serum level of zinc measured at 20 weeks was maintained similarly in each group (P = 0.082), intraprostatic zinc concentrations were statistically different. Group 1 prostates contained the least amount of zinc in both the dorsolateral and ventral lobes at levels of 36.3 and 4.8 microg g(-1), respectively. However, in group 3, zinc levels increased in both lobes to 59.3 and 12.1 microg g(-1), respectively, comparable with that of group 4 (54.5 +/- 14.6 and 14.1 +/- 2.4 microg g(-1)). In spite of these increases in zinc concentration, the prevalence of prostate intraepithelial neoplasm was rather increased in group 3 (53.3% and 46.7%) compared with group 1 (33.3% and 33.3%) in both dorsolateral and ventral prostate lobes. Although prostate intraepithelial neoplasm did not develop in any prostate in group 4, zinc administration did induce prostate intraepithelial neoplasm in group 2 (46.7% and 40.0%). Thus, although high dietary zinc increased intraprostatic zinc concentrations, it promoted, instead of preventing, prostate intraepithelial neoplasm in a murine prostate malignancy induction model. PMID- 20010850 TI - The ubiquitin proteasome system and its involvement in cell death pathways. PMID- 20010851 TI - Small peptide inhibitor of JNKs protects against MPTP-induced nigral dopaminergic injury via inhibiting the JNK-signaling pathway. AB - Increasing evidence suggests that apoptosis may be the mechanism underlying cell death in selective loss of nigral dopaminergic neurons in Parkinson's disease (PD). Previous studies strongly suggested that c-Jun N-terminal kinase (JNK) signaling pathway has a critical role in the animal model with 1-methyl-4-phenyl 1,2,3,6-tetrahydropyridine (MPTP)-induced PD. In this study, we report the inhibitory effect of a peptide designated as Tat-JBD on JNKs activation. The sequence of Tat is corresponding to the cell-membrane transduction domain of human immunodeficiency virus-type 1 (HIV-1) and the sequence of an 11-amino acid peptide is corresponding to the residues of JNK-binding domain (JBD) on JNK interacting protein-1 (JIP-1). Tat-JBD is confirmed to perturb the assembly of JIP-1-JNKs complex, inhibit the activation of JNKs induced by MPTP and consequently diminish the phosphorylation of c-Jun. It also inhibits the phosphorylation of Bcl-2 and the releasing of Bax from Bcl-2/Bax dimmers, sequentially attenuates the translocation of Bax to mitochondria, the release of cytochrome c, the activation of caspase3 and the hydrolyzation of poly-ADP-ribose polymerase. The death of dopaminergic neurons and the loss of dopaminergic axon in the striatum were significantly suppressed by infusion of the peptide Tat-JBD in MPTP-treated mice. Our findings imply that Tat-JBD offers neuroprotection against MPTP injury via inhibiting the JNK-signaling pathway, and may provide a promising therapeutic approach for PD. PMID- 20010852 TI - Rapid quantification of DNA methylation by measuring relative peak heights in direct bisulfite-PCR sequencing traces. AB - Various technologies are currently available to quantify DNA methylation. However, rapid and simple methods for determining the DNA methylation status of CpG sites in genes still remain elusive. In this report, we describe a novel method for the rapid quantification of CpG methylation on the basis of direct bisulfite-PCR sequencing method. According to the principles of bisulfite-PCR, converting unmethylated cytosines to thymine while leaving methylated cytosines unchanged, we regard the CpG site as a SNP and estimate the methylation status of cytosines in the given CG dinucleotides by measuring the ratio of the cytosine peak height to the sum of cytosine and thymine peak heights in automated DNA sequencing traces. Furthermore, we take several effective measures to break through the 'bottleneck' problems that render the routine bisulfite sequencing method unsuitable for quantitative methylation. In comparison with pyrosequencing and bisulfite-cloning sequencing, our method is confirmed to be a simple, high throughput and cost-effective technology for determining the methylation status of specific genes. Accordingly, this novel method is anticipated to be an efficient and economical alternative tool for rapid quantification of methylation patterns in screening large numbers of clinical samples across multiple genes. PMID- 20010853 TI - Morphological differences during in vitro chondrogenesis of bone marrow-, synovium-MSCs, and chondrocytes. AB - Mesenchymal stem cells (MSCs) from a variety of mesenchymal tissue contain common features, but distinguishing properties dependent on their origin are emerging. We investigated morphological differences of human bone marrow-MSCs, synovium MSCs, and chondrocytes during in vitro chondrogenesis. Two hundred thousands cells were pelleted after centrifugation and cultured in chondrogenic media that contained BMP-2, TGF-beta3, and dexamethasone. The pellets were analyzed histologically, immunohistologically, and electron microscopically. Before chondrogenic induction, trypsinized MSCs and chondrocytes looked similar. At day 1, the structure of the three masses was divided into two layers, and the most obvious differences in the three populations were observed in the deep zone. In bone marrow-MSCs, round cells accumulated without intercellular space, and the cells were mainly connected through intermediate junctions. In synovium-MSCs, elongated cells accumulated with small desmosomes and intercellular spaces could occasionally be seen. In chondrocytes, separated oval and polygonal cells connected only in a narrow spotty area through a small desmosome. At day 7, the structure of the three masses was divided into three layers, and the most obvious differences in the three populations were observed in the middle zone. In bone marrow-MSCs, the middle zone consisted of dense smaller cells and apoptotic cells. In synovium-MSCs, the middle zone consisted of dense arrayed wider cells and apoptotic cells. In chondrocytes, the middle zone was acellular without apoptotic cells. At day 21, the morphology of cells and extracellular space became similar in that each cell was located separately with abundant extracellular matrix. The superficial zone was still obvious in bone marrow-MSCs, but hardly seen both in synovium-MSCs and chondrocytes. In this study, we revealed morphological differences of bone marrow-MSCs, synovium-MSCs, and chondrocytes during in vitro chondrogenesis. The most obvious differences in the three populations were observed at day 1 in the deep zone. PMID- 20010855 TI - It's all about patients. PMID- 20010854 TI - ALDH1A1 is a marker for malignant prostate stem cells and predictor of prostate cancer patients' outcome. AB - Prostate cancer (PCa) contains a small population of cancer stem cells (CSCs) that contribute to its initiation and progression. The development of specific markers for identification of the CSCs may lead to new diagnostic strategies of PCa. Increased aldehyde dehydrogenase 1A1 (ALDH1A1) activity has been found in the stem cell populations of leukemia and some solid tumors. The aim of the study was to investigate the stem-cell-related function and clinical significance of the ALDH1A1 in human PCa. ALDEFLUOR assay was used to isolate ALDH1A1(+) cells from PCa cell lines. Stem cell characteristics of the ALDH1A1(+) cells were then investigated by in vitro and in vivo approaches. The ALDH1A1 expression was also analyzed by immunohistochemistry in 18 normal prostate and 163 PCa tissues. The ALDH1A1(+) PCa cells showed high clonogenic and tumorigenic capacities, and serially reinitiated transplantable tumors that resembled histopathologic characteristics and heterogeneity of the parental PCa cells in mice. Immunohistochemical analysis of human prostate tissues showed that ALDH1A1(+) cells were sparse and limited to the basal component in normal prostates. However, in tumor specimens, increased ALDH1A1 immunopositivity was found not only in secretory type cancer epithelial cells but also in neuroendocrine tumor populations. Furthermore, the high ALDH1A1 expression in PCa was positively correlated with Gleason score (P=0.01) and pathologic stage (P=0.01), and inversely associated with overall survival and cancer-specific survival of the patients (P=0.00093 and 0.00017, respectively). ALDH1A1 could be a prostate CSC related marker. Measuring its expression might provide a potential approach to study tumorigenesis of PCa and predict outcome of the disease. PMID- 20010859 TI - The skin as an endocrine target. PMID- 20010860 TI - Exploring the "thyroid-skin connection": concepts, questions, and clinical relevance. AB - The "thyroid-skin connection" has become a hot frontier in dermatoendocrinology. In this context, the pituitary hormone that controls thyroid hormone production, thyrotropin (TSH), caught the attention of skin researchers only after it was reported that the receptor for TSH (TSH-R) is transcribed and translated by selected cultured human skin cell populations and in normal human scalp skin in situ, introducing skin as a nonconventional, peripheral target organ for regulation by TSH. In this issue, Cianfarani et al. present corroborating evidence and raise the old question of whether intracutaneous TSH-R stimulation by autoantibodies contributes to the skin changes seen in autoimmune thyroid disease. This provides a good opportunity to define the questions to be addressed when exploring the thyroid-skin connection. PMID- 20010861 TI - What's in a name?: Heat shock protein 27 and keratinocyte differentiation. AB - In this issue of the Journal, Robitaille and colleagues present data supporting a role for the small heat shock protein (HSP) 27 in keratinocyte terminal differentiation. This adds to the growing literature implicating HSP27 as a regulator of biologic function beyond thermal stress response. PMID- 20010862 TI - Melanoma molecular subtypes: unifying and paradoxical results. AB - In this issue, Hacker and colleagues provide further evidence that molecular subtypes of malignant melanoma may develop along divergent pathways. The authors did not find an association between somatic BRAF-mutant melanoma and germline melanocortin-1 receptor (MC1R) gene status. We discuss this seeming paradox in light of previous studies demonstrating strong associations. PMID- 20010863 TI - Xmrk in medaka: a new genetic melanoma model. AB - Melanoma is the most lethal form of skin cancer, and its incidence is rising rapidly (Chin et al., 2006; Linos et al., 2009). Highly aggressive, metastatic melanoma is notoriously resistant to chemotherapy, and early detection is critical for surgical excision (Gray-Schopfer et al., 2007). A detailed knowledge of the cellular, molecular, and genetic events during melanoma progression is highly relevant to both diagnosis and the development of new therapies. Animal models, such as the one described in this issue by Schartl and colleagues, are important tools for identifying the genetic and environmental factors that contribute to melanoma development, as well as identifying and testing new therapeutic strategies. PMID- 20010864 TI - Sun-sensitizing effects of PKCepsilon shine on multiple mouse strains. AB - Like many skin phenotypes, susceptibility to carcinogenesis is profoundly influenced by genetic background. Sand et al. (this issue) explored the sensitizing effects of epidermal protein kinase C epsilon (PKCepsilon)expression in the hairless SKH-1 mouse strain commonly used in UV carcinogenesis studies. They reported that PKCepsilon overexpression profoundly sensitized these mice to UV skin carcinogenesis and activated oncogenic pathways similar to those reported in FVB/N mice. PMID- 20010865 TI - T-cell post-transplant lymphoproliferative disorder in a patient with chronic idiopathic myelofibrosis following allogeneic PBSC transplantation. PMID- 20010866 TI - Hepatic veno-occlusive disease in children after hematopoietic stem cell transplantation: incidence, risk factors, and outcome. AB - Four hundred and sixty-seven hematopoietic stem cell transplantations (HSCTs) (217 autologous and 250 allogeneic HSCT) were performed in 374 children at four pediatric HSCT centers in Korea from January 2005 to December 2007. Among 467 transplants, veno-occlusive disease (VOD) developed in 72 transplants (15.4%) at a median of 10 days after HSCT. Multivariate analysis showed that BU or TBI containing regimen (P=0.002), VOD prophylaxis without lipo-prostaglandin E1 (PGE1) (P=0.012), number of previous HSCT (P=0.014), and pretransplant serum ferritin (P=0.018) were independent risk factors for developing VOD. Mean serum ferritin levels were significantly higher in HSCT with VOD (2109.6+/-2842.5 ng/ml) than in HSCT without VOD (1315.9+/-1094.4 ng/ml) (P<0.001). The relative risk of death within 100 days of HSCT in transplants with VOD compared with transplants without VOD was 3.39 (confidence interval: 1.78-6.45). Our results suggest that lipo-PGE1 might have a protective effect against the development of VOD, and pretransplant serum ferritin could act as a risk factor for VOD. A larger prospective study is needed to confirm a possible role of lipo-PGE1 and iron chelation therapy in reducing the incidence of VOD. PMID- 20010867 TI - Unrelated cord blood transplantation in pediatric patients: a report from Saudi Arabia. AB - In unrelated cord blood (UCB) transplantation, survival has been shown to correlate with the degree of HLA matching. Thus, to extend transplant access to different ethnic backgrounds, many western UCB transplantation banks now encourage donation from non-Caucasians. Until recently, Saudi Arabia did not have a national UCB bank. In this study we report our experience in UCB transplantation in children using units procured from western cord blood banks. A total of 97 children underwent unrelated UCB transplantation at King Faisal Specialist Hospital and Research Center (KFSHRC), of which 95 were of Arab ethnicity. A total of 30 patients had malignant disorders, 25 patients had non malignant hematological disorders and 42 patients had inborn errors. Conditioning was according to disease, with six patients receiving reduced-intensity regimens. In all, 46 patients received one-Ag-mismatched units and 51 received two-Ag mismatched units. Engraftment occurred in 93% of patients, the 100-TRM was 15%, acute GVHD developed in 20% of patients and chronic GVHD occurred in 9% of patients. The 5-year OS and EFS estimates were 52 and 43%, respectively. The search for UCB transplantation units for Saudi patients in western banks yielded reasonably compatible units for our patients; the results are consistent with published data. Our data are encouraging for UCB transplant programs in countries in which there are no national UCB transplantation banks. PMID- 20010868 TI - Does location matter? Rural vs urban outcomes after blood and marrow transplantation in a population-based Canadian cohort. AB - Specialized health services, such as blood and marrow transplantation (BMT), are usually based in large urban centers. Previous research has suggested that rural patients undergoing BMT have a higher risk of death. We performed a cohort study using data from both the Manitoba BMT Program and the provincial Cancer Registry to determine whether patients from the rural areas would have inferior survival after BMT and whether rural patients have reduced access to BMT. A total of 463 adult Manitobans, who underwent BMT between January 1990 and December 2006, were assessed. We analyzed area of residence (rural vs urban), disease and BMT characteristics, and calculated the OS. Patients undergoing autologous and allogeneic transplants were analyzed separately. When adjusted for gender, age at BMT and year of BMT, area of residence was not a significant predictor of mortality. A relative survival analysis was also conducted, and area of residence was again not a significant predictor of mortality. To measure access to BMT in urban vs rural patients, we evaluated all patients with newly diagnosed Hodgkin's Lymphoma (HL) during this same period. Of 432 Manitobans diagnosed with HL, 182 (42%) were rural and 250 (58%) were urban. In contrast, 69% of patients undergoing transplant for HL were urban. In conclusion, using population-based data from a Canadian province, we were unable to show a survival disadvantage for rural patients after controlling for other variables. BMT utilization in rural populations deserves further study. PMID- 20010869 TI - Enhanced activity of the CREB co-activator Crtc1 in LKB1 null lung cancer. AB - Activation of Crtc1 (also known as Mect1/Torc1) by a t(11;19) chromosomal rearrangement underlies the etiology of malignant salivary gland tumors. As LKB1 is a target for mutational inactivation in lung cancer and was recently shown to regulate hepatic Crtc2/CREB transcriptional activity in mice, we now present evidence suggesting disruption of an LKB1/Crtc pathway in cancer. Although Crtc1 is preferentially expressed in adult brain tissues, we observed elevated levels of steady-state Crtc1 in thoracic tumors. In addition, we show that somatic loss of LKB1 is associated with underphosphorylation of endogenous Crtc1, enhanced Crtc1 nuclear localization and enhanced expression of the Crtc prototypic target gene, NR4A2/Nurr1. Inhibition of NR4A2 was associated with growth suppression of LKB1 null tumors, but showed little effect on LKB1-wildtype cells. These data strengthen the role of dysregulated Crtc as a bona fide cancer gene, present a new element to the complex LKB1 tumorigenic axis, and suggest that Crtc genes may be aberrantly activated in a wider range of common adult malignancies. PMID- 20010870 TI - Copine-III interacts with ErbB2 and promotes tumor cell migration. AB - ErbB2 amplification and overexpression in breast cancer correlates with aggressive disease and poor prognosis. To find novel ErbB2-interacting proteins, we used stable isotope labeling of amino acids in cell culture followed by peptide affinity pull-downs and identified specific binders using relative quantification by mass spectrometry. Copine-III, a member of a Ca(2+)-dependent phospholipid-binding protein family, was identified as binding to phosphorylated Tyr1248 of ErbB2. In breast cancer cells, Copine-III requires Ca(2+) for binding to the plasma membrane, where it interacts with ErbB2 upon receptor stimulation, an interaction that is dependent on receptor activity. Copine-III also binds receptor of activated C kinase 1 and colocalizes with phosphorylated focal adhesion kinase at the leading edge of migrating cells. Importantly, knockdown of Copine-III in T47D breast cancer cells causes a decrease in Src kinase activation and ErbB2-dependent wound healing. Our data suggest that Copine-III is a novel player in the regulation of ErbB2-dependent cancer cell motility. In primary breast tumors, high CPNE3 RNA levels significantly correlate with ERBB2 amplification. Moreover, in an in situ tissue microarray analysis, we detected differential protein expression of Copine-III in normal versus breast, prostate and ovarian tumors, suggesting a more general role for Copine-III in carcinogenesis. PMID- 20010871 TI - The promyelocytic leukemia zinc-finger gene, PLZF, is frequently downregulated in malignant mesothelioma cells and contributes to cell survival. AB - DNA copy number analysis was performed, using single-nucleotide polymorphism mapping arrays, to fine map genomic imbalances in human malignant mesothelioma (MM) cell lines derived from primary tumors. Chromosomal losses accounted for the majority of genomic imbalances. All 22 cell lines examined showed homozygous deletions of 9p21.3, centering at the CDKN2A/ARF and CDKN2B loci. Other commonly underrepresented segments included 1p36, 1p22, 3p21-22, 4q13, 4q34, 11q23, 13q12 13, 14q32, 15q15, 18q12, and 22q12, each observed in 55-90% of cell lines. Focal deletions of 11q23 encompassed the transcriptional repressor gene promyelocytic leukemia zinc finger (PLZF), which was validated by analysis of genomic DNA using real-time polymerase chain reaction (PCR). Semi-quantitative RT-PCR and immunoblot analysis revealed that PLZF is greatly downregulated in MM cell lines compared with non-malignant mesothelial cells. Ectopic expression of PLZF in PLZF deficient MM cells resulted in decreased cell viability, reduced colony formation, as well as increased apoptosis, the latter based on results of various cell death assays and the observation of increased cleavage of caspase 3, PARP, and Mcl-1. These data indicate that deletions of PLZF are a common occurrence in MM and that downregulation of PLZF may contribute to MM pathogenesis by promoting cell survival. PMID- 20010872 TI - Src family tyrosine kinases-driven colon cancer cell invasion is induced by Csk membrane delocalization. AB - The nonreceptor tyrosine kinases of the Src family (SFK) are frequently deregulated in human colorectal cancer (CRC), and they have been implicated in tumour growth and metastasis. How SFK are activated in this cancer has not been clearly established. Here, we show that the SFK-dependent invasion is induced by inactivation of the negative regulator C-terminal Src kinase, Csk. While the level of Csk was inconsistent with SFK activity in colon cancer cells, its membrane translocation, needed for efficient regulation of membrane-localized SFK activity, was impaired. Accordingly, Csk downregulation did not affect SFK oncogenic activity in these cells, whereas expression of a membrane-localized form of this kinase affected their invasive activity. Downregulation of the transmembrane and rafts-localized Csk-binding protein/phosphoprotein associated with glycosphingolipid-enriched microdomain (PAG), was instrumental for the cytoplasmic accumulation of Csk. Re-expression of PAG in cells from late-stage CRC inhibited SFK invasive activity in a Csk-dependent manner. Conversely, inactivation of its residual expression in early-stage CRC cells promoted SFK invasive activity. Finally, this mechanism was specific to CRC as Csk coupling to SFK was readily detected in breast cancer cells. Therefore, Csk mis-localization defines a novel mechanism for SFK oncogenic activation in CRC cells. PMID- 20010873 TI - Complete deletion of Apc results in severe polyposis in mice. AB - The adenomatous polyposis coli (APC) gene product is mutated in the vast majority of human colorectal cancers. APC negatively regulates the WNT pathway by aiding in the degradation of beta-catenin, which is the transcription factor activated downstream of WNT signaling. APC mutations result in beta-catenin stabilization and constitutive WNT pathway activation, leading to aberrant cellular proliferation. APC mutations associated with colorectal cancer commonly fall in a region of the gene termed the mutation cluster region and result in expression of an N-terminal fragment of the APC protein. Biochemical and molecular studies have revealed localization of APC/Apc to different sub-cellular compartments and various proteins outside of the WNT pathway that associate with truncated APC/Apc. These observations and genotype-phenotype correlations have led to the suggestion that truncated APC bears neomorphic and/or dominant-negative function that support tumor development. To analyze this possibility, we have generated a novel allele of Apc in the mouse that yields complete loss of Apc protein. Our studies reveal that whole-gene deletion of Apc results in more rapid tumor development than the APC multiple intestinal neoplasia (Apc(Min)) truncation. Furthermore, we found that adenomas bearing truncated Apc had increased beta catenin activity when compared with tumors lacking Apc protein, which could lead to context-dependent inhibition of tumorigenesis. PMID- 20010874 TI - Smad2 and Smad3 have opposing roles in breast cancer bone metastasis by differentially affecting tumor angiogenesis. AB - Transforming growth factor (TGF)-beta can suppress and promote breast cancer progression. How TGF-beta elicits these dichotomous functions and which roles the principle intracellular effector proteins Smad2 and Smad3 have therein, is unclear. Here, we investigated the specific functions of Smad2 and Smad3 in TGF beta-induced responses in breast cancer cells in vitro and in a mouse model for breast cancer metastasis. We stably knocked down Smad2 or Smad3 expression in MDA MB-231 breast cancer cells. The TGF-beta-induced Smad3-mediated transcriptional response was mitigated and enhanced by Smad3 and Smad2 knockdown, respectively. This response was also seen for TGF-beta-induced vascular endothelial growth factor (VEGF) expression. TGF-beta induction of key target genes involved in bone metastasis, were found to be dependent on Smad3 but not Smad2. Strikingly, whereas knockdown of Smad3 in MDA-MB-231 resulted in prolonged latency and delayed growth of bone metastasis, Smad2 knockdown resulted in a more aggressive phenotype compared with control MDA-MB-231 cells. Consistent with differential effects of Smad knockdown on TGF-beta-induced VEGF expression, these opposing effects of Smad2 versus Smad3 could be directly correlated with divergence in the regulation of tumor angiogenesis in vivo. Thus, Smad2 and Smad3 differentially affect breast cancer bone metastasis formation in vivo. PMID- 20010875 TI - NF-kappaB signaling mediates the induction of MTA1 by hepatitis B virus transactivator protein HBx. AB - Metastasis-associated protein 1 (MTA1), a master chromatin modifier, has been shown to regulate cancer progression and is widely upregulated in human cancer, including hepatitis B virus-associated hepatocellular carcinomas (HCCs). Here we provide evidence that hepatitis B virus transactivator protein HBx stimulates the expression of MTA1 but not of MTA2 or MTA3. The underlying mechanism of HBx stimulation of MTA1 involves HBx targeting of transcription factor nuclear factor (NF)-kappaB and the recruitment of HBx/p65 complex to the NF-kappaB consensus motif on the relaxed MTA1 gene chromatin. We also discovered that MTA1 depletion in HBx-expressing cells severely impairs the ability of HBx to stimulate NF kappaB signaling and the expression of target proinflammatory molecules. Furthermore, the presence of HBx in HBx-infected HCCs correlated well with increased MTA1 and NF-kappaB-p65. Collectively, these findings revealed a previously unrecognized integral role of MTA1 in HBx stimulation of NF-kappaB signaling and consequently, the expression of NF-kappaB targets gene products with functions in inflammation and tumorigenesis. PMID- 20010876 TI - Ezrin mediates c-Myc actions in prostate cancer cell invasion. AB - The forced overexpression of c-Myc in mouse prostate and in normal human prostate epithelial cells results in tumor transformation with an invasive phenotype. How c-Myc regulates cell invasion is poorly understood. In this study, we have investigated the interplay of c-Myc and androgens in the regulation of prostate cancer cell invasion. We found that c-Myc induces cell invasion and anchorage independent growth by regulating ezrin protein expression in the presence of androgens. The activity of the ezrin promoter is controlled by androgens through c-Myc, which binds to a phylogenetically conserved E-Box located in the proximal promoter region. Besides, we also show that ezrin is an important regulator of c Myc protein levels. These effects are achieved through androgen-induced changes in ezrin phosphorylation, which results in the regulation of downstream signals. These downstream signals involve the modulation of Akt and GSK-3beta activity resulting in increased c-Myc protein synthesis and inhibition of its degradation. In summary, we have shown a key role for ezrin as a mediator of c-Myc-induced tumorigenesis in prostate cancer cells. PMID- 20010879 TI - Determinants of kidney dysfunction: is vasopressin a new player in the arena? AB - Copeptin is a marker of endogenous vasopressin secretion. Population-based data of the PREVEND Study show that men and women with high plasma copeptin have a highly prevalent microalbuminuria in addition to the low level of hydration typical of persons with high vasopressin secretion. The association of plasma copeptin with microalbuminuria suggests that vasopressin might have a role in kidney dysfunction via effects on the permselectivity of the glomerulus and/or on tubular albumin reabsorption. PMID- 20010880 TI - Epidemiology of atherosclerotic renal artery stenosis as a mirror of prospective trials. AB - Diagnosis and treatment of atherosclerotic renal artery stenosis (ARAS) in the elderly is a never-ending challenge for every nephrologist. Kalra et al. report that adjusted hazard ratios for ARAS increased threefold from 1992 to 2004 in 16 million United States Renal Data System participants aged 66 years or older. However, numbers of revascularizations showed a biphasic pattern with a declining number since 1999. These exciting data have to be discussed with the knowledge of recent prospective trials. PMID- 20010881 TI - HIV-1 entry into human podocytes is mediated through lipid rafts. PMID- 20010882 TI - Dialysis patients: vulnerable group of patients. PMID- 20010885 TI - Nomenclature of the Oxford classification of IgA nephropathy: do we need to be careful? PMID- 20010886 TI - The case: milky ascites is not always chylous. PMID- 20010887 TI - Infected glomerulus. PMID- 20010889 TI - Brain weight in males is correlated with DNA methylation at IGF2. PMID- 20010890 TI - The reaction norm in gene x environment interaction. PMID- 20010891 TI - The missing link; the significance of urocortin 1/urocortin 2 in the modulation of the dorsal raphe serotoninergic system. PMID- 20010888 TI - The CRF system, stress, depression and anxiety-insights from human genetic studies. AB - A concatenation of findings from preclinical and clinical studies support a preeminent function for the corticotropin-releasing factor (CRF) system in mediating the physiological response to external stressors and in the pathophysiology of anxiety and depression. Recently, human genetic studies have provided considerable support to several long-standing hypotheses of mood and anxiety disorders, including the CRF hypothesis. These data, reviewed in this report, are congruent with the hypothesis that this system is of paramount importance in mediating stress-related psychopathology. More specifically, variants in the gene encoding the CRF(1) receptor interact with adverse environmental factors to predict risk for stress-related psychiatric disorders. In-depth characterization of these variants will likely be important in furthering our understanding of the long-term consequences of adverse experience. PMID- 20010892 TI - Predictable chronic mild stress improves mood, hippocampal neurogenesis and memory. AB - Maintenance of neurogenesis in adult hippocampus is important for functions such as mood and memory. As exposure to unpredictable chronic stress (UCS) results in decreased hippocampal neurogenesis, enhanced depressive- and anxiety-like behaviors, and memory dysfunction, it is believed that declined hippocampal neurogenesis mainly underlies the behavioral and cognitive abnormalities after UCS. However, the effects of predictable chronic mild stress (PCMS) such as the routine stress experienced in day-to-day life on functions such as mood, memory and hippocampal neurogenesis are unknown. Using FST and EPM tests on a prototype of adult rats, we demonstrate that PCMS (comprising 5 min of daily restraint stress for 28 days) decreases depressive- and anxiety-like behaviors for prolonged periods. Moreover, we illustrate that decreased depression and anxiety scores after PCMS are associated with ~1.8-fold increase in the production and growth of new neurons in the hippocampus. Additionally, we found that PCMS leads to enhanced memory function in WMT as well as NORT. Collectively, these findings reveal that PCMS is beneficial to adult brain function, which is exemplified by increased hippocampal neurogenesis and improved mood and cognitive function. PMID- 20010893 TI - X chromosome and suicide. AB - Suicide completion rates are significantly higher in males than females in most societies. Although gender differences in suicide rates have been partially explained by environmental and behavioral factors, it is possible that genetic factors, through differential expression between genders, may also help explain gender moderation of suicide risk. This study investigated X-linked genes in suicide completers using a two-step strategy. We first took advantage of the genetic structure of the French-Canadian population and genotyped 722 unrelated French-Canadian male subjects, of whom 333 were suicide completers and 389 were non-suicide controls, using a panel of 37 microsatellite markers spanning the entire X chromosome. Nine haplotype windows and several individual markers were associated with suicide. Significant results aggregated primarily in two regions, one in the long arm and another in the short arm of chromosome X, limited by markers DXS8051 and DXS8102, and DXS1001 and DXS8106, respectively. The second stage of the study investigated differential brain expression of genes mapping to associated regions in Brodmann areas 8/9, 11, 44 and 46, in an independent sample of suicide completers and controls. Six genes within these regions, Rho GTPase activating protein 6, adaptor-related protein complex 1 sigma 2 subunit, glycoprotein M6B, ribosomal protein S6 kinase 90 kDa polypeptide 3, spermidine/spermine N(1)-acetyltransferase 1 and THO complex 2, were found to be differentially expressed in suicide completers. PMID- 20010894 TI - Dysbindin-1 genotype effects on emotional working memory. AB - We combined functional imaging and genetics to investigate the behavioral and neural effects of a dysbindin-1 (DTNBP1) genotype associated with the expression level of this important synaptic protein, which has been implicated in schizophrenia. On a working memory (WM) task for emotional faces, participants with the genotype related to increased expression showed higher WM capacity for happy faces compared with the genotype related to lower expression. Activity in several task-related brain areas with known DTNBP1 expression was increased, including hippocampal, temporal and frontal cortex. Although these increases occurred across emotions, they were mostly observed in areas whose activity correlated with performance for happy faces. This suggests effects of variability in DTNBP1 on emotion-specific WM capacity and region-specific task-related brain activation in humans. Synaptic effects of DTNBP1 implicate that altered dopaminergic and/or glutamatergic neurotransmission may be related to the increased WM capacity. The combination of imaging and genetics thus allows us to bridge the gap between the cellular/molecular and systems/behavioral level and extend the cognitive neuroscience approach to a comprehensive biology of cognition. PMID- 20010895 TI - Modelling disrupted-in-schizophrenia 1 loss of function in human neural progenitor cells: tools for molecular studies of human neurodevelopment and neuropsychiatric disorders. PMID- 20010896 TI - Diabetic nephropathy: a disorder of oxygen metabolism? AB - Chronic hypoxia induces sequential abnormalities in oxygen metabolism (for example, oxidative stress, nitrosative stress, advanced glycation, carbonyl stress, endoplasmic reticulum stress) in the kidneys of individuals with diabetes. Identification of these abnormalities improves our understanding of therapeutic benefits that can be achieved with antihypertensive agents, the control of hyperglycemia and/or hyperinsulinemia and the dietary correction of obesity. Key to the body's defense against hypoxia is hypoxia-inducible factor, the activity of which is modulated by prolyl hydroxylases (PHDs)-oxygen sensors whose inhibition may prove therapeutic. Renal benefits of small-molecule PHD inhibitors have been documented in several animal models, including those of diabetic nephropathy. Three different PHD isoforms have been identified (PHD1, PHD2 and PHD3) and their respective roles have been delineated in knockout mouse studies. Unfortunately, none of the current inhibitors is specific for a distinct PHD isoform. Nonspecific inhibition of PHDs might induce adverse effects, such as those associated with PHD2 inhibition. Specific disruption of PHD1 induces hypoxic tolerance, without angiogenesis and erythrocytosis, through the reprogramming of basal oxygen metabolism and decreased generation of oxidative stress in hypoxic mitochondria. A specific PHD1 inhibitor might, therefore, offer a novel therapy for abnormal oxygen metabolism not only in the diabetic kidney, but also in other diseases for which hypoxia is a final, common pathway. PMID- 20010897 TI - Epidemiology of peritoneal dialysis: a story of believers and nonbelievers. AB - In 2008, an estimated 1.77 million patients worldwide received dialysis. Of these patients, 1.58 million were treated with hemodialysis and approximately 190,000 received peritoneal dialyisis. In a global comparison of treatment methods for renal failure, therefore, hemodialysis clearly dominates. In this Review, we compare the epidemiology of peritoneal dialysis with that of hemodialysis and describe some of the major differences in the global utilization of the two dialysis modalities. These differences can largely be explained by a number of nonmedical, mainly economic factors, but also by educational and psychological factors. To reverse the current trends, the implementation of suitable reimbursement strategies for peritoneal dialysis is needed as well as increased investment in the training of young nephrology fellows and in education programs for patients and other non-nephrological health-care providers. To achieve these goals, academic and nonacademic training centers, which often consider peritoneal dialysis to be a low-level priority, must invest in research and training related to peritoneal dialysis. PMID- 20010898 TI - Intraperitoneal therapy for peritoneal tumors: biophysics and clinical evidence. AB - In patients with tumors confined to the peritoneal cavity, there is established pharmacokinetic and tumor biology-related evidence that intraperitoneal drug administration is advantageous. Three large randomized trials in patients with stage III ovarian cancer who underwent optimal cytoreduction have demonstrated a significant survival benefit when intraperitoneal chemotherapy was added to systemic therapy. Although intraperitoneal therapy is associated with locoregional toxic effects, recent trials suggest that with some modification of the local delivery methods this approach is safe in 80% of patients in an ambulatory setting. Surgical cytoreduction immediately followed by intraoperative hyperthermic intraperitoneal chemoperfusion (HIPEC) ensures intraperitoneal delivery of the drug to all peritoneal surfaces and the advantages of combined hyperthermia to be exploited. An increasing number of centers are initiating this multimodality therapy in ovarian cancer and colorectal cancer. Clearly, intraperitoneal drug delivery is an important adjunct to surgery and systemic chemotherapy in selected patients. The optimal drug, dose and schedule for intraperitoneal delivery, the exact role of added HIPEC compared with cytoreduction alone, and the potential role of HIPEC in ovarian cancer and peritoneal mesothelioma are still undefined. Several randomized controlled trials addressing these uncertainties have been initiated. PMID- 20010899 TI - Genomic markers for decision making: what is preventing us from using markers? AB - The advent of novel genomic technologies that enable the evaluation of genomic alterations on a genome-wide scale has significantly altered the field of genomic marker research in solid tumors. Researchers have moved away from the traditional model of identifying a particular genomic alteration and evaluating the association between this finding and a clinical outcome measure to a new approach involving the identification and measurement of multiple genomic markers simultaneously within clinical studies. This in turn has presented additional challenges in considering the use of genomic markers in oncology, such as clinical study design, reproducibility and interpretation and reporting of results. This Review will explore these challenges, focusing on microarray-based gene-expression profiling, and highlights some common failings in study design that have impacted on the use of putative genomic markers in the clinic. Despite these rapid technological advances there is still a paucity of genomic markers in routine clinical use at present. A rational and focused approach to the evaluation and validation of genomic markers is needed, whereby analytically validated markers are investigated in clinical studies that are adequately powered and have pre-defined patient populations and study endpoints. Furthermore, novel adaptive clinical trial designs, incorporating putative genomic markers into prospective clinical trials, will enable the evaluation of these markers in a rigorous and timely fashion. Such approaches have the potential to facilitate the implementation of such markers into routine clinical practice and consequently enable the rational and tailored use of cancer therapies for individual patients. PMID- 20010900 TI - Salivary habituation to food stimuli in successful weight loss maintainers, obese and normal-weight adults. AB - OBJECTIVE: Research shows that slower habituation of salivary responses to food stimuli is related to greater energy intake and that obese (Ob) individuals habituate slower than those of normal weight (NW). No study has examined habituation rates in weight loss maintainers (WLMs) who have reduced from obese to normal weight, relative to those who are Ob or NW. DESIGN: Salivation to two baseline water trials and 10 lemon-flavored lollipop trials were studied in 14 WLMs, 15 Ob and 18 NW individuals comparable in age, gender and ethnicity. Linear mixed models were used to compare WLMs with Ob and NW groups. RESULTS: Salivation in the WLM and NW groups decreased significantly (for both P <0.005) across trials, indicative of habituation. Salivary responses in the Ob group did not habituate (P=0.46). When compared with Ob group, WLMs showed a quicker reduction in salivation (P<0.05). WLM and NW groups did not differ in habituation rate (P=0.49). CONCLUSIONS: WLMs have habituation rates that are comparable to NW individuals without previous history of obesity, and show quicker habituation than those who are currently obese. These results suggest that physiological responses to food may 'normalize' with successful weight loss maintenance. PMID- 20010901 TI - Obesity-induced impairment of endothelium-dependent vasodilation in Tunisian women. AB - OBJECTIVE: It is now well recognized that obesity is a major public health concern, and its prevalence has tremendously increased worldwide over the last decades, including Tunisia. As obesity is associated with cardiovascular diseases, the purpose of this study was to investigate the effect of obesity on forearm skin blood flow (FSBF) response to acetylcholine (Ach), an endothelium dependent vasodilator, in Tunisian women over a wide range of body mass indices (BMIs). SUBJECTS: One hundred and eighty healthy women with an average age of 34+/-6 years, an average height of 162+/-7 cm and an average weight of 78+/-19 kg participated in this investigation. The mean BMIs of the 60 lean, 50 overweight and 70 obese subjects were 22.1+/-0.3, 27.7+/-0.2 and 38.4+/-0.7 kg m(-2), respectively. MEASUREMENTS: The FSBF was measured non-invasively using a laser Doppler flowmeter in response to local infusion of a cumulative dose of Ach. RESULTS: After adjusting for age, the mean response of FSBF to Ach was significantly greater in lean (1168%+/-78) than in overweight (643%+/-38) and obese subjects (323%+/-18) (P=0.002; P<0.0001, respectively), suggesting a reduction of the endothelium-dependent nitric oxide (NO) release by obesity. Our regression analysis also revealed that the maximum FSBF response to Ach (that is, its efficacy) was inversely correlated with BMI, waist and hip circumferences (r= 0.994, P=0.002; r=-0.2, P<0.0001, and r=-0.321, P=0.001, respectively). CONCLUSION: Our data demonstrate a reduction of skin vasodilatory reserve in obese patients and suggest a defect of both endothelial-dependent relaxation and wall compliance associated with obesity. PMID- 20010902 TI - Pancreatic steatosis: harbinger of type 2 diabetes in obese rodents. AB - OBJECTIVE: The aim of this study was to determine if the fat accumulation in the exocrine pancreas fat of obese Zucker diabetic fatty (ZDF) rodents, like that in their endocrine pancreas, precedes the onset of type 2 diabetes mellitus (T2DM). As the fat content of whole pancreas, but not islets, can now be measured in humans by magnetic resonance spectroscopy (MRS), such measurements could be used as a predictor of impending T2DM and an indication for preventive intervention. ANIMALS: Obese ZDF (fa/fa) rats and lean (+/+) controls on a 6% fat diet were killed at time points from 6 to 16 weeks and total pancreatic fat was measured biochemically and electronmicroscopic examination of tissue for fat droplets was carried out. RESULTS: Compared to lean ZDF controls, pancreatic fat was elevated above lean controls from 6 to 16 weeks of age, peaking at 10 weeks of age when hyperglycemia first appeared. The pancreatic profile of fat content in whole pancreas paralleled that of islets. Electronmicroscopic examination identified the acinar location of the fat droplets and ruled out a major contribution of intrapancreatic adipocytes. CONCLUSION: The almost identical pattern of triglyceride overaccumulation in the exocrine and endocrine pancreas of obese rodents before the onset of T2DM suggests that MRS of the human pancreas might predict T2DM in obese subjects and permit timely interventions to prevent the disease. PMID- 20010903 TI - The influence of body mass index, age and gender on current illness: a cross sectional study. AB - CONTEXT: Obesity poses a significant health risk, but health risk is not equivalent to actual health status. Further, age and gender might alter the effect of body weight on physical health. OBJECTIVE: To determine the relationship between body mass index (BMI), age, gender and current health status. DESIGN: Data from the 1988-1994, 2003-2004 and 2005-2006 National Health & Nutrition Examination Surveys were weighted to represent the US population. BMI, age, gender and current medication use were analyzed in a sample-adjusted 9071 women and 8880 men. MAIN OUTCOME MEASURES: The percentage of participants taking medication and the total number of medications taken. RESULTS: In both the 1988-1994 and 2003-2006 data sets, with few exceptions, medication loads did not increase significantly in overweight compared with normal-weight people. Medication loads increased significantly in obese compared with normal-weight people aged 40+, but only marginally at 25-39 years. Medication loads were higher in women than men, but significantly less so in people aged 55-70. CONCLUSIONS: First, medication loads, a measure of current health status, were increased in obese compared with the normal-weight people, but the effect was mainly at ages over 40 years. In addition, BMI category contributed less to medication loads at ages 25-39 than in older groups. Second, there was little difference in current health status in normal-weight versus overweight people at all ages. Finally, higher medication loads in women than men are more apparent in younger than older people. Although obesity does not substantially affect current health in young people, it is likely that the increased medication loads in obese compared with normal-weight older people originates at least in part from an increased BMI starting at a younger age. Thus, age, gender and onset of high BMI all require consideration when using BMI to assess current health status. PMID- 20010904 TI - Stevioside inhibits atherosclerosis by improving insulin signaling and antioxidant defense in obese insulin-resistant mice. AB - OBJECTIVE: Stevioside is a non-caloric natural sweetener that does not induce a glycemic response, making it attractive as sweetener to diabetics and others on carbohydrate-controlled diets. Obesity is frequently associated with insulin resistance and increased inflammation and oxidative stress. Therefore, we investigated its effects on insulin resistance, inflammation and oxidative stress related to atherosclerosis in obese insulin-resistant mice. RESEARCH DESIGN: Twelve-week-old mice were treated with stevioside (10 mg kg(-1), n=14) or placebo (n=20) for 12 weeks. RESULTS: Stevioside had no effect on weight and triglycerides, but lowered glucose and insulin. Stevioside treatment improved adipose tissue maturation, and increased glucose transport, insulin signaling and antioxidant defense in white visceral adipose tissues. Together, these increases were associated with a twofold increase of adiponectin. In addition, stevioside reduced plaque volume in the aortic arch by decreasing the macrophage, lipid and oxidized low-density lipoprotein (ox-LDL) content of the plaque. The higher smooth muscle cell-to-macrophage ratio was indicative for a more stable plaque phenotype. The decrease in ox-LDL in the plaque was likely due to an increase in the antioxidant defense in the vascular wall, as evidenced by increased Sod1, Sod2 and Sod3. Circulating adiponectin was associated with improved insulin signaling and antioxidant defense in both the adipose tissue and the aorta of stevioside-treated mice. CONCLUSION: Stevioside treatment was associated with improved insulin signaling and antioxidant defense in both the adipose tissue and the vascular wall, leading to inhibition of atherosclerotic plaque development and inducing plaque stabilization. PMID- 20010905 TI - Inaccuracies in food and physical activity diaries of obese subjects: complementary evidence from doubly labeled water and co-twin assessments. AB - OBJECTIVE: To study whether eating or physical-activity (PA) habits differ between obese and non-obese monozygotic (MZ) co-twins independent of genetic effects. METHODS: Rare MZ pairs discordant for obesity (n=14, body mass index difference 5.2+/-1.8 kg m(-2)) and weight-concordant control pairs (n=10, 1.0+/ 0.7 kg m(-2)), identified through a population-based registry of 24-28-year-old twins (n=658 MZ pairs), completed 3-day food and PA diaries and eating behavior questionnaires. Each twin was asked to compare his/her own eating and PA patterns with the co-twin's behavior by structured questionnaires. Accuracy of energy intake was validated by doubly labeled water. RESULTS: Non-obese co-twins consistently reported that their obese twin siblings ate more food overall, consumed less healthy foods and exercised less than the non-obese co-twins do. However, no differences in energy intake (9.6+/-1.0 MJ per day vs 9.8+/-1.1 MJ per day, respectively) in the food diaries or in the mean PA level (1.74+/-0.02 vs 1.79+/-0.04, respectively) in the PA diaries were found between obese and non obese co-twins. A considerable underreporting of energy intake (3.2+/-1.1 MJ per day, P=0.036) and overreporting of PA (1.8+/-0.8 MJ per day, P=0.049) was observed in the obese, but not in the non-obese co-twins. CONCLUSIONS: On the basis of rare MZ twin pairs discordant for obesity, the co-twin assessments confirmed substantial differences in eating and PA behavior between obese and non obese persons. These may be overlooked in population studies using food and PA diaries because of considerable misreporting by the obese. PMID- 20010907 TI - Brown-Sequard syndrome due to penetrating injury by an iron fence point. AB - STUDY DESIGN: Case report. OBJECTIVES: To present an unusual type of penetrating objects causing Brown-Sequard syndrome (BSS) and its clinical character. SETTING: Department of Orthopaedic Surgery, Shanghai Changzheng Hospital, Second Military Medical University, Shanghai, PR China. METHODS: A 54-year-old man fell from a height of 4 m onto an iron fence, and a sharp iron fence point penetrated the right side of his back. He developed left-sided BSS. Both X-ray film and computed tomography scanning of the thoracic spine showed a right vertebral plate of Th5 fracture and metal fragments inclining through the posterior and left lateral of the spinal canal. Emergency decompressive laminectomy and removal of the foreign metal piece were performed. No improvement in neurological function was observed 10 days after surgery, and thus hyperbaric oxygen treatment was initiated twice a day for the next 1 month. RESULTS: Forty days after surgery, his bladder function returned to normal. The motor deficit had regressed and he could walk without assistance 70 days after the operation. One year later, his lower extremity functions recovered almost completely, except for slight numbness on the right side. CONCLUSION: As far as we know, on the basis of existing literature, the injury mechanism to BSS by a sharp iron fence point has not been reported so far. The satisfactory recovery after injury may in part be attributed to timely surgery and continuing hyperbaric oxygen treatment. PMID- 20010906 TI - Influence of maternal educational level on the association between the rs3809508 neuromedin B gene polymorphism and the risk of obesity in the HELENA study. AB - OBJECTIVE: Neuromedin B (NMB) is a bombesin-like peptide, which inhibits food intake and modulates stress-related behaviour. An NMB gene polymorphism (P73T) has been earlier associated with obesity and abnormal eating behaviour in adults. METHODS: The association between four NMB polymorphisms and obesity-related phenotypes was investigated in the Healthy Lifestyle in Europe by Nutrition in Adolescence cross-sectional study (n=1144, 12-17-year-old European adolescents). This population was genotyped for the NMB rs1107179, rs17598561, rs3809508 and rs1051168 (P73T) polymorphisms. Obesity was defined according to Cole et al. (BMJ 2000; 320:1240-1243) criteria; eating behaviour was assessed by the Eating Behaviour and Weight Problems Inventory for Children (EWI-C) and the food choices and preferences questionnaires. Familial socioeconomic status (SES) was assessed through the parents' educational level. RESULTS: Only the genotype distribution of rs3809508 differed according to obesity status, as the TT genotype was more frequent in obese than in non-obese adolescents (8.6% vs 3.1%, P=0.05; adjusted odds ratio for obesity (95% confidence interval): 2.85 (1.11-7.31), P=0.03). Moreover, TT subjects had higher body mass index (22.8+/-4.4 kg m(-2) vs 21.3+/ 3.7 kg m(-2), P=0.02), waist circumference (75.8+/-9.7 cm vs 72.2+/-9.3 cm, P=0.006), waist-to-hip ratio (0.84+/-0.14 vs 0.79+/-0.07, P<0.0001) and waist-to height ratio (0.47+/-0.06 vs 0.44+/-0.55, P=0.002) than C allele carriers. The effects of this single nucleotide polymorphism on all anthropometric values were influenced by the maternal SES, in that a low maternal educational level aggravated the phenotype of adolescents carrying the TT genotype (interactions: P<0.02). No association with EWI-C scores was found, although sweet craving was a more frequent cause of between-meal food intake in TT subjects than in C allele carriers (24.3% vs 9.2%, P=0.01). CONCLUSION: In European adolescents, the TT genotype of the NMB rs3809508 polymorphism was associated with a higher risk of obesity. Moreover, the effects of this polymorphism on anthropometric values were influenced by the maternal educational level. PMID- 20010908 TI - Changes in pubococcygeus muscle tension in the pelvic floor of rats after spinal cord injury. AB - STUDY DESIGN: The purpose of this study was to explore the changes in pelvic floor muscle tension at different times after suprasacral spinal cord injury (SS) and sacral cord injury (SC), and learn more about pelvic floor dysfunction (PFD) after spinal cord injury (SCI). METHOD: A total of 70 healthy female Sprague Dawley (SD) rats, weighing between 250 and 280 g, were randomly divided into seven groups with 10 rats in each group, which included five SS groups (3 days and 2, 4, 8 and 12 weeks after injury), one SC group (4 weeks after injury) and one normal group. Muscle tension, including muscle compliance, and contraction activity elicited using electrostimulation under two initial lengths were measured at different time points. RESULTS: (1) Muscle compliance decreased within 4 weeks in the SS group (P>0.05), began to increase at 8 weeks (P<0.05) and reached the peak at 12 weeks, which were all lower than the normal level; (2) contraction activity under both initial lengths tended to decrease within 4 weeks, peak at 8 weeks (P<0.05) and decline again at 12 weeks , which were all lower than that of the normal group as well; and (3) the SC group showed similar compliance with the normal group (P>0.05) and less contraction activity when compared with other SS groups (P<0.05). CONCLUSION: Measurement for the compliance and contraction activity of pubococcygeus indicates the changes from decreasing to increasing after suprasacral cord injury, and similar compliance combined with rather low contraction activity compared with the normal group after sacral cord injury, both of which are in accordance with PFD after spinal cord injury. PMID- 20010909 TI - Evaluation of the physical activity scale for individuals with physical disabilities in people with spinal cord injury. AB - STUDY DESIGN: Cross-sectional study. OBJECTIVES: To evaluate the physical activity scale for individuals with physical disabilities (PASIPD) in people with spinal cord injury (SCI). SETTING: Eight Dutch rehabilitation centers with a specialized SCI unit. METHODS: The PASIPD was examined by comparing group scores of people with different personal (age, gender and body mass index) and lesion characteristics (level (paraplegia/tetraplegia), completeness, time since injury (TSI)) in 139 persons with SCI 1 year after discharge from in-patient rehabilitation. Relationships between PASIPD scores and measures of activities (wheelchair skills, Utrecht Activity List, mobility range and social behavior subscales of the SIP68) and fitness (peak oxygen uptake, peak power output and muscular strength) were determined. RESULTS: Persons with tetraplegia had significantly lower PASIPD scores than those with paraplegia (P<0.02). Persons with longer TSI had lower PASIPD scores than persons with shorter TSI (P<0.03). PASIPD scores showed moderate correlations with activities (0.36-0.51, P<0.01) and weak-to-moderate correlations with fitness parameters (0.25-0.36, P<0.05). CONCLUSION: In a fairly homogeneous group of persons with SCI, 1 year after in patient rehabilitation, the PASIPD showed weak-to-moderate relationships with activity and fitness parameters. There seems to be a limited association between self-reported activity level and fitness in people with SCI. PMID- 20010910 TI - Transplantation of a combination of autologous neural differentiated and undifferentiated mesenchymal stem cells into injured spinal cord of rats. AB - STUDY DESIGN: The use of stem cells for functional recovery after spinal cord injury. OBJECTIVE: The aim of this study was to evaluate the effects of a combination of autologous undifferentiated and neural-induced bone marrow mesenchymal stem cells (MSCs) on behavioral improvement in rats after inducing spinal cord injury and comparing with transplantation of undifferentiated and neural-induced MSCs alone. SETTING: The study was conducted at the department of Clinical Sciences, Faculty of Veterinary Medicine, University of Tehran, Tehran, Iran. METHODS: The spinal cord was injured by contusion using a Fogarty embolectomy catheter at the T8-T9 level of the spinal cord, and autologous MSCs were transplanted into the center of the developing lesion cavity, 3 mm cranial and 3 mm caudal to the cavity, at 7 days after induction of spinal cord compression injury. RESULTS: At 5 weeks after transplantation, the presence of transplanted cells was detected in the spinal cord parenchyma using immunohistochemistry analysis. In all treatment groups (differentiated, undifferentiated and mix), there was less cavitation than lesion sites in the control group. The Basso-Beattie-Bresnahan (BBB) score was significantly higher in rats transplanted with a combination of cells and in rats transplanted with neural-induced MSCs alone than in undifferentiated and control rats. CONCLUSION: Pre-differentiation of MSCs to neuron-like cells has a very important role in achieving the best results for functional improvement. PMID- 20010911 TI - Pain after paraplegia: a survey in India. AB - STUDY DESIGN: A postal survey. OBJECTIVE: To ascertain the incidence of pain in individuals with paraplegia in India and to associate it with demographic characteristics. SETTING: India. METHOD: The study was done by means of a questionnaire. This questionnaire was mailed to the identified individuals (n=600) on the addresses obtained from the medical records section of hospitals and from various organizations. Data analysis was done by using non-parametric tests of association. RESULTS: The return rate was 46% (276/600). Fifty-seven percent of individuals complained of pain. Of this, pain in the back and chest ranked the highest (30.1%), followed by pain below the level of lesion (6.9%), pain in the shoulder and upper limb (4.7%) and neck (0.4%). Fifteen percent of individuals complained of pain at multiple sites. We found a significant association of pain with age, duration since injury and ambulation. CONCLUSION: More than half of the subjects complained of pain in the study. Pain was found to be associated with age, duration since injury and ambulation. As pain has a dramatic effect on a subjects' quality of life, there is a need to evaluate it in detail and treat accordingly with preventive, rehabilitative or surgical procedures. PMID- 20010912 TI - Differential requirements for IL-4/STAT6 signalling in CD4 T-cell fate determination and Th2-immune effector responses. AB - Improved analytical tools have revealed that the development and expression of a Th2 immune response can be broken down into distinct stages with respect to the cytokine microenvironment that is required. Although IL-4 and its STAT6 signalling pathway are critical for the expression of Th2 effector immune responses in peripheral tissues such as the skin, lung and gut, IL-4 and STAT6 signalling are not required for the initial generation of IL-4-producing Th2 cells in the lymph node. This finding reveals that we have yet to identify the key cytokine or microenvironment that stimulates the development of this most intriguing CD4(+) T-helper subset and emphasises the tissue specificity and timing of IL-4/STAT6-dependent Th2 effector responses. PMID- 20010913 TI - Unique features of NK cell development during ontogeny revealed in studies of RAG 1-deficient mice. PMID- 20010914 TI - Association of polymorphisms of the cannabinoid receptor (CNR1) and fatty acid amide hydrolase (FAAH) genes with heroin addiction: impact of long repeats of CNR1. AB - Alterations in expression of a cannabinoid receptor (CNR1, CB1), and of fatty acid amide hydrolase (FAAH) that degrades endogenous ligands of CB1, may contribute to the development of addiction. The 385C>A in the FAAH gene and six polymorphisms of CNR1 were genotyped in former heroin addicts and control subjects (247 Caucasians, 161 Hispanics, 179 African Americans and 19 Asians). In Caucasians, long repeats (>or=14) of 18087-18131(TAA)(8-17) were associated with heroin addiction (P=0.0102). Across three ethnicities combined, a highly significant association of long repeats with heroin addiction was found (z=3.322, P=0.0009). Point-wise significant associations of allele 1359A (P=0.006) and genotype 1359AA (P=0.034) with protection from heroin addiction were found in Caucasians. Also in Caucasians, the genotype pattern, 1359G>A and -6274A>T, was significantly associated with heroin addiction experiment wise (P=0.0244). No association of FAAH 385C>A with heroin addiction was found in any group studied. PMID- 20010915 TI - Complement and its role in innate and adaptive immune responses. AB - The complement system plays a crucial role in the innate defense against common pathogens. Activation of complement leads to robust and efficient proteolytic cascades, which terminate in opsonization and lysis of the pathogen as well as in the generation of the classical inflammatory response through the production of potent proinflammatory molecules. More recently, however, the role of complement in the immune response has been expanded due to observations that link complement activation to adaptive immune responses. It is now appreciated that complement is a functional bridge between innate and adaptive immune responses that allows an integrated host defense to pathogenic challenges. As such, a study of its functions allows insight into the molecular underpinnings of host-pathogen interactions as well as the organization and orchestration of the host immune response. This review attempts to summarize the roles that complement plays in both innate and adaptive immune responses and the consequences of these interactions on host defense. PMID- 20010916 TI - Heterogeneity and plasticity of T helper cells. AB - CD4 T helper (Th) cells play critical roles in adaptive immune responses. They recruit and activate other immune cells including B cells, CD8 T cells, macrophages, mast cells, neutrophils, eosinophils and basophils. Based on their functions, their pattern of cytokine secretion and their expression of specific transcription factors, Th cells, differentiated from naive CD4 T cells, are classified into four major lineages, Th1, Th2, Th17 and T regulatory (Treg) cells, although other Th lineages may exist. Subsets of the same lineage may express different effector cytokines, reside at different locations or give rise to cells with different fates, whereas cells from different lineages may secrete common cytokines, such as IL-2, IL-9 and IL-10, resulting in massive heterogeneity of the Th cell population. In addition, the pattern of cytokine secretion may switch from that of one lineage toward another under certain circumstances, suggesting that Th cells are plastic. Tregs are also more heterogeneous and plastic than were originally thought. In this review, we summarize recent reports on heterogeneity and plasticity of Th cells, and discuss potential mechanisms and implications of such features that Th cells display. PMID- 20010917 TI - The use of inhibitors to study endocytic pathways of gene carriers: optimization and pitfalls. AB - Nonviral gene complexes can enter mammalian cells through different endocytic pathways. For efficient optimization of the gene carrier it is important to profile its cellular uptake, because this largely determines its intracellular processing and subsequent transfection efficiency. Most of the current information on uptake of these gene-delivery vehicles is based on data following the use of chemical inhibitors of endocytic pathways. Here, we have performed a detailed characterization of four commonly used endocytosis inhibitors [chlorpromazine, genistein, methyl-beta-cyclodextrin (MbetaCD), and potassium depletion] on cell viability and endocytosis in five well-described cell lines. We found that chlorpromazine and to a lesser extent MbetaCD significantly decreased cell viability of some cell lines even after short incubation periods and at concentrations that are routinely used to inhibit endocytosis. Through analyzing the uptake and subcellular distribution of two fluorescent endocytic probes transferrin and lactosylceramide (LacCer) that are reported to enter cells via clathrin-dependent (CDE) and clathrin-independent (CIE) mechanisms, respectively, we showed poor specificity of these agents for inhibiting distinct endocytic pathways. Finally, we demonstrate that any inhibitory effects are highly cell line dependent. Overall, the data question the significance of performing endocytosis studies with these agents in the absence of very stringent controls. PMID- 20010918 TI - Comparative transduction efficiency of AAV vector serotypes 1-6 in the substantia nigra and striatum of the primate brain. AB - Vectors derived from adeno-associated virus (AAV) are promising candidates for neural cell transduction in vivo because they are nonpathogenic and achieve long term transduction in the central nervous system. AAV serotype 2 (AAV2) is the most widely used AAV vector in clinical trials based largely on its ability to transduce neural cells in the rodent and primate brain. Prior work in rodents suggests that other serotypes might be more efficient; however, a systematic evaluation of vector transduction efficiency has not yet been performed in the primate brain. In this study, AAV viral vectors of serotypes 1-6 with an enhanced green-fluorescent protein (GFP) reporter gene were generated at comparable titers, and injected in equal amounts into the brains of Chlorocebus sabaeus. Vector injections were placed in the substantia nigra (SN) and the caudate nucleus (CD). One month after injection, immunohistochemistry for GFP was performed and the total number of GFP+ cells was calculated using unbiased stereology. AAV5 was the most efficient vector, not only transducing significantly more cells than any other serotype, but also transducing both NeuN+ and glial-fibrillary-acidic protein positive (GFAP+) cells. These results suggest that AAV5 is a more effective vector than AAV2 at delivering potentially therapeutic transgenes to the nigrostriatal system of the primate brain. PMID- 20010919 TI - Ten-day sequential therapy of Helicobacter pylori infection in Thailand. AB - OBJECTIVES: Antimicrobial resistance has decreased eradication rates of Helicobacter pylori worldwide. The objective of this study was to determine whether a sequential therapy regimen is effective in eradicating H. pylori in adults with nonulcer dyspepsia or peptic ulcer disease in Thailand. METHODS: A total of 115 patients with dyspepsia or peptic ulcer were enrolled in the study. (14)C-urea breath test, upper endoscopy, rapid urease test, bacterial culture, and antibiotic resistance assessment were conducted during the course of the treatment. In all, 115 patients underwent a 10-day sequential regimen, which consisted of lansoprazole (30 mg) plus amoxicillin (1 g) twice a day for 5 days, then lansoprazole (30 mg) with metronidazole (500 mg) twice a day, and clarithromycin (1,000 mg) once a day for another 5 consecutive days. Successful eradication was evaluated by negative urea breath test at least 4 weeks after stopping treatment. RESULTS: Successful eradication was achieved in 106 of 115 patients (95%). All patients completed the treatment, without any dropouts. Mild adverse effects included headache and palpitations. The prevalence rate of clarithromycin-resistant H. pylori was found to be 6.1%. CONCLUSIONS: The 10-day sequential treatment for H. pylori is well tolerated and provides a high eradication rate. This regimen can overcome the emergence of antibiotic resistance and may have a role as a first-line treatment for H. pylori infection in Thailand. PMID- 20010920 TI - Dyspeptic symptoms and endoscopic findings in the community: the Loiano Monghidoro study. AB - OBJECTIVES: We aimed to evaluate the prevalence of endoscopic findings and their association with dyspeptic symptoms in the community. METHODS: A total of 1,533 inhabitants of two villages were invited to participate in a cross-sectional survey, and 1,033 were recruited. Participants underwent a validated dyspepsia questionnaire, upper gastrointestinal endoscopy, and a (13)C-urea breath test. RESULTS: Endoscopic findings were present in 17.6% of asymptomatic subjects and in 27.4% of those with dyspeptic symptoms. The prevalence of esophagitis and Barrett's esophagus in subjects with dyspeptic symptoms and without prominent reflux symptoms was 8.1 and 1.5%, respectively, and was similar to that of asymptomatic subjects (8.5 and 0.7%, respectively). Esophagitis was significantly associated with dyspeptic symptoms only in subjects with concomitant prominent reflux symptoms. Peptic ulcer (PU) was present in 8.8% of subjects with dyspeptic symptoms without reflux symptoms and similarly in 9.4% of those with prominent reflux symptoms. Subjects with dyspeptic symptoms and concomitant prominent reflux symptoms had an increased risk of having an underlying PU (odds ratio 2.74, 95% confidence interval 1.30-5.78). CONCLUSIONS: Almost three-quarters of subjects with dyspeptic symptoms do not have endoscopic findings and, in addition, esophagitis may not be the cause of dyspeptic symptoms in subjects without prominent reflux symptoms. PU may be the cause of dyspeptic symptoms in a subgroup of subjects with prominent reflux symptoms. PMID- 20010921 TI - Mucosal serotonin signaling is altered in chronic constipation but not in opiate induced constipation. AB - OBJECTIVES: Changes in mucosal serotonin (5-HT) signaling have been detected in a number of functional and inflammatory disorders of the gastrointestinal (GI) tract. This study was undertaken to determine whether chronic constipation (CC) is associated with disordered 5-HT signaling and to evaluate whether constipation caused by opiate use causes such changes. METHODS: Human rectal biopsy samples were obtained from healthy volunteers, individuals with idiopathic CC, and individuals taking opiate medication with or without occurrence of constipation. EC cells were identified by 5-HT immunohistochemistry. 5-HT content and release levels were determined by enzyme immunoassay, and mRNA levels for the synthetic enzyme tryptophan hydroxylase-1 (TpH-1) and serotonin-selective reuptake transporter (SERT) were assessed by quantitative real-time reverse transcription PCR. RESULTS: CC was associated with increases in TpH-1 transcript, 5-HT content, and 5-HT release under basal and stimulated conditions, whereas EC cell numbers and SERT transcript levels were not altered. No changes in these elements of 5-HT signaling were detected in opiate-induced constipation (OIC). CONCLUSIONS: These findings demonstrate that CC is associated with a pattern of altered 5-HT signaling that leads to increased 5-HT availability but does not involve a decrease in SERT expression. It is possible that increased 5-HT availability due to increased synthesis and release contributes to constipation due to receptor desensitization. Furthermore, the finding that elements of 5-HT signaling were not altered in the mucosa of individuals with OIC indicates that constipation as a condition does not lead to compensatory changes in 5-HT synthesis, release, or signal termination. PMID- 20010922 TI - Investigation of the associations between low-dose serum perfluorinated chemicals and liver enzymes in US adults. AB - OBJECTIVES: Perfluorinated chemicals (PFCs) have been largely used for years in a variety of products worldwide. However, the toxic effect of PFCs on exposure to the liver in the general population has not yet been determined. METHODS: In this study, 2,216 adults (18 years of age or older) were recruited in a National Health and Nutrition Examination Survey (NHANES) in 1999-2000 and 2003-2004 to determine the relationship between serum level of PFCs and the levels of liver enzymes. The data were adjusted for all other confounding variants. RESULTS: After performing mathematical analysis, we determined when serum log perfluorooctanoic acid (PFOA) increases in one unit, the serum alanine aminotransferase (ALT) concentration (U/l) increases by 1.86 units (95% confidence interval (CI), 1.24-2.48; P=0.005), and the serum log-gamma glutamyltransferase (GGT) concentration (U/l) is 0.08 unit higher (95% CI, 0.05 0.11; P=0.019). The association between PFOA and liver enzymes was more evident in obese subjects, as well as subjects with insulin resistance and/or metabolic syndromes. When dividing the serum PFOA into quartiles in the fully adjusted models in subjects with a body mass index>or=30 kg/m2, the ALT level trend across the serum PFOA quartiles was significant (P=0.003). CONCLUSIONS: On the basis of these data, we conclude that a higher serum concentration of PFOA may cause liver enzymes to increase abnormally in the general population, particularly in obese individuals. Further studies are warranted to clarify the casual relationship between PFCs and these liver enzymes. PMID- 20010923 TI - CIMP status of interval colon cancers: another piece to the puzzle. AB - OBJECTIVES: Colon cancers diagnosed in the interval after a complete colonoscopy may occur due to limitations of colonoscopy or due to the development of new tumors, possibly reflecting molecular and environmental differences in tumorigenesis resulting in rapid tumor growth. In a previous study from our group, interval cancers (colon cancers diagnosed within 5 years of a complete colonoscopy) were almost four times more likely to demonstrate microsatellite instability (MSI) than non-interval cancers. In this study we extended our molecular analysis to compare the CpG island methylator phenotype (CIMP) status of interval and non-interval colorectal cancers and investigate the relationship between the CIMP and MSI pathways in the pathogenesis of interval cancers. METHODS: We searched our institution's cancer registry for interval cancers, defined as colon cancers that developed within 5 years of a complete colonoscopy. These were frequency matched in a 1:2 ratio by age and sex to patients with non interval cancers (defined as colon cancers diagnosed on a patient's first recorded colonoscopy). Archived cancer specimens for all subjects were retrieved and tested for CIMP gene markers. The MSI status of subjects identified between 1989 and 2004 was known from our previous study. Tissue specimens of newly identified cases and controls (between 2005 and 2006) were tested for MSI. RESULTS: There were 1,323 cases of colon cancer diagnosed over the 17-year study period, of which 63 were identified as having interval cancer and matched to 131 subjects with non-interval cancer. Study subjects were almost all Caucasian men. CIMP was present in 57% of interval cancers compared to 33% of non-interval cancers (P=0.004). As shown previously, interval cancers were more likely than non-interval cancers to occur in the proximal colon (63% vs. 39%; P=0.002), and have MSI 29% vs. 11%, P=0.004). In multivariable logistic regression model, proximal location (odds ratio (OR) 1.85; 95% confidence interval (CI) 1.01-3.8), MSI (OR 2.7; 95% CI 1.1-6.8) and CIMP (OR 2.41; 95% CI 1.2-4.9) were independently associated with interval cancers. CIMP was associated with interval cancers independent of MSI status. There was no difference in 5-year survival between the two groups. CONCLUSIONS: Interval cancers are more likely to arise in the proximal colon and demonstrate CIMP, which suggests there may be differences in biology between these and non-interval CRC. Additional studies are needed to determine whether interval cancers arise as a result of missed lesions or accelerated neoplastic progression. PMID- 20010924 TI - The combined measurement of total serum IgG and IgG4 may increase diagnostic sensitivity for autoimmune pancreatitis without sacrificing specificity, compared with IgG4 alone. AB - OBJECTIVES: Until now, there was no international consensus on the diagnostic criteria for autoimmune pancreatitis (AIP). As for serologic criteria, the HISORt criteria use elevated immunoglobulin (Ig) G4 alone, whereas the Asian diagnostic criteria include elevations of total IgG or IgG4, or the presence of autoantibodies. This study was mainly aimed at determining whether the combined measurement of total IgG and IgG4 could increase the diagnostic sensitivity for AIP while maintaining specificity, compared with IgG4 alone. Another aim was to determine the utility of autoantibodies to diagnose AIP. METHODS: We prospectively measured total serum IgG and IgG4 together in 82 consecutive patients with AIP, and seropositivity was defined as elevation of either total IgG or IgG4. To evaluate specificity in the differentiation of AIP from pancreatic cancer, total serum IgG and IgG4 were prospectively measured in 110 patients with pancreatic cancer. Also, the detection rates of antinuclear antibody (ANA) and rheumatoid factor (RF) were retrospectively reviewed in patients with AIP. RESULTS: In patients with AIP, the sensitivity of IgG4 (> or = 135 mg/dl) was 52.5% (43/82), significantly higher than that (46.3%, 38/82) of total IgG (> or = 1,800 mg/dl) (P<0.05). The sensitivity of combined measurement of total IgG and IgG4 for AIP was 68.3% (56/82), significantly higher than that of IgG4 alone (P<0.05). The specificity of total IgG and IgG4 in the differentiation of AIP from pancreatic cancer was 96.4 and 99.1%, respectively. The specificity of combined measurement of total IgG and IgG4 was 95.5%, and it was not significantly different from that of IgG4 alone (P=0.125). The sensitivity of ANA (> or = 1:80) and RF was 24.4% (19/78) and 20.3% (13/64), respectively. All but one patient who had positive results for ANA or RF also showed elevations of either total IgG or IgG4, respectively. CONCLUSIONS: The combined measurement of total serum IgG and IgG4 may increase diagnostic sensitivity without sacrificing specificity, compared with IgG4 alone. However, the measurement of ANA or RF may show no additional benefit when combined with total serum IgG and IgG4. PMID- 20010925 TI - The impact of thiopurines on the risk of surgical recurrence in patients with Crohn's disease after first intestinal surgery. AB - OBJECTIVES: Smoking and a lack of immunosuppressive (IS) therapy are considered risk factors for intestinal surgery in Crohn's disease (CD). Good evidence for the latter is lacking. The objective of this study was to evaluate the impact of thiopurine treatment on surgical recurrence in patients after first intestinal resection for CD and its possible interaction with smoking. METHODS: Data on 326 patients after first intestinal resection were retrieved retrospectively, and subjects were grouped according to their postoperative exposure to thiopurines. Treatment with either azathioprine (AZA) or 6-mercaptopurine (6-MP) was recorded on 161 patients (49%). Smoking status was assessed by directly contacting the patients. RESULTS: Surgical recurrence occurred in 151/326 (46.3%) patients after a median time of 71 (range 3-265) months. Cox regression revealed a significant reduction of re-operation rate in patients treated with AZA/6-MP for > or = 36 months as compared with patients treated for 3-35 months, for less than 3 months, and to those without postoperative treatment with AZA/6-MP (P=0.004). Cox regression analysis revealed treatment with thiopurines for > or = 36 months (hazard ratio (HR) 0.41; 95% confidence interval (CI) 0.23-0.76, P=0.004) and smoking (HR 1.6; 95% CI 1.14-2.4, P=0.008) as independent predictors for surgical recurrence. Furthermore, longer duration of disease tended to be protective (HR 0.99; 95% CI 0.99-1.0, P=0.067). CONCLUSIONS: Long-term maintenance treatment with AZA/6-MP reduces the risk of surgical recurrence in patients with CD. We also identified smoking as a risk factor for surgical recurrence. PMID- 20010926 TI - Synchronous culture of Plasmodium falciparum at high parasitemia levels. AB - This protocol describes a method for preparing cultures of Plasmodium falciparum synchronized at any intraerythrocytic stage. Using this method, around 60% parasitized cells may be obtained. On the basis of Trager and Jensen's original continuous culture method, our approach relies on the use of fresh human blood not older than 2 weeks, a low hematocrit between 0.8 and 1.5%, a starting frozen inoculum of 10% ring-stage parasitemia, human serum replaced with AlbuMAX I and alternating sorbitol and Percoll synchronization methods to shorten the cycle window to 4-6 h and reduce sorbitol toxicity. From our synchronized high parasite density cultures, 3-5 ml of infected red blood cells can be obtained in 1 week, corresponding to 1.2 mg of total parasite protein per ml of harvested culture. On the basis of the variables parasitemia and packed cell volume, we provide an equation to accurately calculate the amount of complete medium required every 24 h corrected for the cycle stage and capacity of the culture flask. Ten days suffice to complete the protocol from a frozen stock of parasites. PMID- 20010927 TI - High-yield cell-free protein production from P-gel. AB - Cell-free systems represent a promising approach to quickly and easily produce preparative amounts of proteins. However, it is still challenging to obtain high volumetric yields (>mg ml(-1)) of proteins from the present cell-free systems. This protocol presents a cell-free protein synthesis method using a novel DNA gel that dramatically increases protein yield compared with current systems. This protein-producing gel (termed 'P-gel system' or 'P-gel'), which consists of genes as part of the gel scaffolding, can produce mg ml(-1) amounts of functional proteins. This protocol describes steps pertaining to plasmid design, fabrication of P-gel molds, formation of P-gel micropads and cell-free protein expression with an expected yield of up to 5 mg ml(-1) of functional Renilla luciferase (Rluc). This entire process can take 1-3 d, depending on the desired quantity of protein. PMID- 20010928 TI - High-throughput haplotype determination over long distances by haplotype fusion PCR and ligation haplotyping. AB - When combined with haplotype fusion PCR (HF-PCR), ligation haplotyping is a robust, high-throughput method for empirical determination of haplotypes, which can be applied to assaying both sequence and structural variation over long distances. Unlike alternative approaches to haplotype determination, such as allele-specific PCR and long PCR, HF-PCR and ligation haplotyping do not suffer from mispriming or template-switching errors. In this method, HF-PCR is used to juxtapose DNA sequences from single-molecule templates, which contain single nucleotide polymorphisms (SNPs) or paralogous sequence variants (PSVs) separated by several kilobases. HF-PCR uses an emulsion-based fusion PCR, which can be performed rapidly and in a 96-well format. Subsequently, a ligation-based assay is performed on the HF-PCR products to determine haplotypes. Products are resolved by capillary electrophoresis. Once optimized, the procedure can be performed quickly, taking a day and a half to generate phased haplotypes from genomic DNA. PMID- 20010929 TI - Efficient expression of tyrosine-sulfated proteins in E. coli using an expanded genetic code. AB - Tyrosine sulfation is an important post-translational modification that occurs in higher eukaryotes and is involved in cell-cell communication, viral entry and adhesion. We describe a protocol for the heterologous expression of selectively tyrosine-sulfated proteins in Escherichia coli through the use of an expanded genetic code that co-translationally inserts sulfotyrosine in response to the amber nonsense codon, TAG. The components required for this process, an orthogonal aminoacyl-tRNA synthetase specific for sulfotyrosine and its cognate orthogonal tRNA that recognizes the amber codon, are encoded on the plasmid pSUPAR6-L3-3SY, and their use, along with a simple chemical synthesis of sulfotyrosine, are outlined in this protocol. Specifically, the gene for a protein of interest is mutated such that the codon corresponding to the desired location of tyrosine sulfate is TAG. Co-transformation of an expression vector containing this gene and pSUPAR6-L3-3SY into an appropriate E. coli strain allows the overexpression of the site-specifically sulfated protein with high efficiency and fidelity. The resulting protein contains tyrosine sulfate at any location specified by a TAG codon, making this method significantly simpler and more versatile than competing methods such as in vitro enzymatic sulfation, chemical sulfation and peptide synthesis. Once the proper expression vectors are cloned, our protocol should allow the production of the desired sulfated proteins in <1 week. PMID- 20010930 TI - Simultaneous analysis of reactive oxygen species and reduced glutathione content in living cells by polychromatic flow cytometry. AB - Reactive oxygen species (ROS) are continuously produced in the cell as a consequence of aerobic metabolism, and are controlled by several antioxidant mechanisms. An accurate measurement of ROS is essential to evaluate the redox status of the cell, or the effects of molecules with the pro-oxidant or antioxidant activity. Here we report a cytofluorimetric technique for measuring simultaneously, at the single-cell level, hydrogen peroxide and superoxide anion, reduced glutathione (a main intracellular antioxidant) and cell viability. The staining is performed with the fluorescent dyes 2',7'-dichlorodihydrofluorescein diacetate (H2DCFH-DA), hydroethidine (HE), monobromobimane (MBB) and TO-PRO-3. This analysis is possible with new-generation flow cytometers equipped with several light sources (in our case, four lasers and an UV lamp), which excite different fluorochromes. This approach is extremely useful to study the balance between ROS content and antioxidants in cells receiving different stimuli, and to analyze the relationship between oxidative stress and cell death. PMID- 20010932 TI - Determination of gene expression patterns using in situ hybridization to Drosophila testes. AB - We describe a whole-mount RNA in situ hybridization (ISH) method optimized for detection of the cellular and subcellular distributions of specific mRNA within Drosophila testes and male genital tract. Digoxygenin (dig)-labeled antisense RNA probes are in vitro transcribed from a template synthesized by (RT)-PCR; the probe length is reduced by hydrolysis. Testes and male genital tracts are dissected from adult flies, fixed and processed for hybridization. Both probe and fixed testes can be stored before use. Extensive post-hybridization washing reduces the background. Detection is through alkaline phosphatase-conjugated anti dig antibodies followed by a color reaction. This protocol is suitable for low medium throughput applications with parallel processing of 2-48 samples, and takes 4-5 d to complete. We have used this protocol, which is similar to other RNA ISH protocols, but optimized for whole-mount Drosophila testes, to document the expression of about 1,000 genes in Drosophila melanogaster male genital tract. PMID- 20010931 TI - Protocols to detect senescence-associated beta-galactosidase (SA-betagal) activity, a biomarker of senescent cells in culture and in vivo. AB - Normal cells can permanently lose the ability to proliferate when challenged by potentially oncogenic stress, a process termed cellular senescence. Senescence associated beta-galactosidase (SA-betagal) activity, detectable at pH 6.0, permits the identification of senescent cells in culture and mammalian tissues. Here we describe first a cytochemical protocol suitable for the histochemical detection of individual senescent cells both in culture and tissue biopsies. The second method is based on the alkalinization of lysosomes, followed by the use of 5-dodecanoylaminofluorescein di-beta-D-galactopyranoside (C12FDG), a fluorogenic substrate for betagal activity. The cytochemical method takes about 30 min to execute, and several hours to a day to develop and score. The fluorescence methods take between 4 and 8 h to execute and can be scored in a single day. The cytochemical method is applicable to tissue sections and requires simple reagents and equipment. The fluorescence-based methods have the advantages of being more quantitative and sensitive. PMID- 20010933 TI - Global identification of protein kinase substrates by protein microarray analysis. AB - Herein, we describe a protocol for the global identification of in vitro substrates targeted by protein kinases using protein microarray technology. Large numbers of fusion proteins tagged at their carboxy-termini are purified in 96 well format and spotted in duplicate onto amino-silane-coated slides in a spatially addressable manner. These arrays are incubated in the presence of purified kinase and radiolabeled ATP, and then washed, dried and analyzed by autoradiography. The extent of phosphorylation of each spot is quantified and normalized, and proteins that are reproducibly phosphorylated in the presence of the active kinase relative to control slides are scored as positive substrates. This approach enables the rapid determination of kinase-substrate relationship on a proteome-wide scale, and although developed using yeast, has since been adapted to higher eukaryotic systems. Expression, purification and printing of the yeast proteome require about 3 weeks. Afterwards, each kinase assay takes approximately 3 h to perform. PMID- 20010934 TI - Targeted mutagenesis in zebrafish using customized zinc-finger nucleases. AB - Zebrafish mutants have traditionally been obtained by using random mutagenesis or retroviral insertions, methods that cannot be targeted to a specific gene and require laborious gene mapping and sequencing. Recently, we and others have shown that customized zinc-finger nucleases (ZFNs) can introduce targeted frame-shift mutations with high efficiency, thereby enabling directed creation of zebrafish gene mutations. Here we describe a detailed protocol for constructing ZFN expression vectors, for generating and introducing ZFN-encoding RNAs into zebrafish embryos and for identifying ZFN-generated mutations in targeted genomic sites. All of our vectors and methods are compatible with previously described Zinc-Finger Consortium reagents for constructing engineered zinc-finger arrays. Using these methods, zebrafish founders carrying targeted mutations can be identified within 4 months. PMID- 20010935 TI - Production of compartmented cultures of rat sympathetic neurons. AB - The compartmented culture, in which primary neurons plated in a proximal compartment send their axons under silicone grease barriers and into left and right distal compartments, has enhanced the experimental capabilities of neuronal cultures. Treatments can be applied separately to cell bodies/proximal axons or distal axons, and cell bodies/proximal axons and distal axons can be separately harvested and analyzed. Distal axons can be axotomized, and the neurons can be studied while their axons regenerate. Construction of the culture dishes requires 3 h for 48 cultures, and preparing the neurons also requires 3 h. Compartmented cultures provide enough cellular material for biochemical analyses such as immunoblotting. The uses of compartmented cultures have included studies of neurotrophic factor retrograde signaling, axonal transport, and axonal protein and lipid biosynthesis. Here we focus on sympathetic neurons cultured from neonatal rats and provide protocols for the production and some of the uses of compartmented cultures. PMID- 20010936 TI - Investigating endothelial invasion and sprouting behavior in three-dimensional collagen matrices. AB - Seeding a monolayer of primary human endothelial cells on the surface of a polymerized three-dimensional collagen matrix in the presence of pro-angiogenic stimuli allows manipulation and analysis of rapid sprouting responses. This protocol is useful for elucidating incompletely defined intracellular mechanisms downstream of pro-angiogenic factors that regulate sprout formation and initiation, and can also be used to test the efficacy of pro-and anti-angiogenic compounds. We present protocols to culture endothelial cells, prepare three dimensional collagen matrices and quantify and image rapid endothelial sprouting responses (24 h). This protocol can be carried out using either type I or type II collagen matrices with primary endothelial cells isolated from macrovascular and microvascular sources of varying species. PMID- 20010937 TI - EOS lentiviral vector selection system for human induced pluripotent stem cells. AB - Generation of induced pluripotent stem (iPS) cells from patients has exciting applications for studying molecular mechanisms of diseases, screening drugs and ultimately for use in cell therapies. However, the low efficiency and heterogeneous nature of reprogramming is a major impediment to the generation of personalized iPS cell lines. We reported in Nature Methods (6, 370-376, 2009) the first selection system to enrich for reprogrammed human iPS cells. Using a lentiviral vector that specifically expresses the enhanced green fluorescence protein and puromycin resistance genes in pluripotent stem cells, it is now possible to mark and enrich for human iPS cell colonies expressing endogenous pluripotency markers. In this study, we describe a detailed protocol for the production of the pluripotent state-specific lentiviral vector and the selection system for the induction of healthy and disease-specific human iPS cells. Overall, preparation of the selection system takes 2 weeks, and the generation of human iPS cells takes approximately 2 months. PMID- 20010938 TI - Agrobacterium-mediated transformation of friable embryogenic calli and regeneration of transgenic cassava. AB - Agrobacterium-mediated transformation of friable embryogenic calli (FEC) is the most widely used method to generate transgenic cassava plants. However, this approach has proven to be time-consuming and can lead to changes in the morphology and quality of FEC, influencing regeneration capacity and plant health. Here we present a comprehensive, reliable and improved protocol, taking approximately 6 months, that optimizes Agrobacterium-mediated transformation of FEC from cassava model cultivar TMS60444. We cocultivate the FEC with Agrobacterium directly on the propagation medium and adopt the extensive use of plastic mesh for easy and frequent transfer of material to new media. This minimizes stress to the FEC cultures and permits a finely balanced control of nutrients, hormones and antibiotics. A stepwise increase in antibiotic concentration for selection is also used after cocultivation with Agrobacterium to mature the transformed FEC before regeneration. The detailed information given here for each step should enable successful implementation of this technology in other laboratories, including those being established in developing countries where cassava is a staple crop. PMID- 20010939 TI - Pre-clinical evaluation of cyclin-dependent kinase 2 and 1 inhibition in anti estrogen-sensitive and resistant breast cancer cells. AB - BACKGROUND: Cellular proliferation, driven by cyclin-dependent kinases (CDKs) and their cyclin partners, is deregulated in cancer. Anti-estrogens, such as tamoxifen, antagonise estrogen-induced ERalpha transactivation of cyclin D1, resulting in reduced CDK4/6 activity, p27(Kip1)-mediated inhibition of CDK2 and growth arrest. We hypothesised that direct inhibition of CDK2 and CDK1 may overcome the major clinical problem of anti-estrogen resistance. METHODS: The cellular effects of CDK2/1 siRNA knockdown and purine-based CDK2/1 inhibitors, NU2058 and NU6102, were measured in anti-estrogen-sensitive and resistant breast cancer cell lines. RESULTS: CDK2 knockdown caused G1 accumulation, whereas CDK1 depletion caused G2/M slowing, and dual CDK1/2 depletion resulted in further G2/M accumulation and cell death in both anti-estrogen-sensitive and resistant cells, confirming CDK2 and CDK1 as targets for breast cancer therapy. In contrast to tamoxifen, which only affected hormone-sensitive cells, NU2058 and NU6102 reduced CDK2-mediated phosphorylation of pRb, E2F transcriptional activity and proliferation, ultimately resulting in cell death, in both anti-estrogen sensitive and resistant cells. Both drugs caused G2/M arrest, reflective of combined CDK2/1 knockdown, with a variable degree of G1 accumulation. CONCLUSION: These studies confirm the therapeutic potential of CDK2 and CDK1 inhibitors for cancer therapy, and support their use as an alternative treatment for endocrine resistant breast cancer. PMID- 20010940 TI - Hypoxia potentiates Notch signaling in breast cancer leading to decreased E cadherin expression and increased cell migration and invasion. AB - BACKGROUND: Epithelial-to-mesenchymal transition (EMT) is associated with decreased adhesion and acquisition of metastatic potential of breast cancer cells. Epithelial-to-mesenchymal transition is mediated, in part, by two transcription repressors, Snail and Slug, that are known to be targets of the Notch signaling pathway, and JAGGED1-induced Notch activation increases EMT. However, the events that lead to increased Notch activity during EMT of breast cancer cells are unknown. METHODS: The accumulation of hypoxia inducible factors (HIFs) under hypoxia was detected by western blot analysis, and their effects on Notch signaling were measured by an in vitro Notch reporter assay. The expression of Notch target genes under hypoxia was tested by real-time PCR. The knockdown of HIF-1alpha was mediated by retroviral delivery of shRNA. The expression of Slug and Snail under hypoxia was measured by real-time PCR. Breast cancer cell migration and invasion under hypoxia were tested with cell migration and invasion kits. RESULTS: Hypoxia increased the expression of Notch target genes such as HES1 and HEY1 in breast cancer cells, as was expression of Notch receptors and ligands. The mechanism is likely to involve the accumulation of HIF-1alpha and HIF-2alpha in these cells by hypoxia, which synergised with the Notch co activator MAML1 in potentiating Notch activity. Hypoxia inducible factor-1alpha was found to bind to HES1 promoter under hypoxia. Knockdown of HIF-1alpha with shRNA inhibited both HES1 and HEY1 expression under hypoxia. Hypoxia increased the expression of Slug and Snail, and decreased the expression of E-cadherin, hallmarks of EMT. Notch pathway inhibition abrogated the hypoxia-mediated increase in Slug and Snail expression, as well as decreased breast cancer cell migration and invasion. CONCLUSION: Hypoxia-mediated Notch signaling may have an important role in the initiation of EMT and subsequent potential for breast cancer metastasis. PMID- 20010941 TI - Prostate cancer cells modulate osteoblast mineralisation and osteoclast differentiation through Id-1. AB - BACKGROUND: Id-1 is overexpressed in and correlated with metastatic potential of prostate cancer. The role of Id-1 in this metastatic process was further analysed. METHODS: Conditioned media from prostate cancer cells, expressing various levels of Id-1, were used to stimulate pre-osteoclast differentiation and osteoblast mineralisation. Downstream effectors of Id-1 were identified. Expressions of Id-1 and its downstream effectors in prostate cancers were studied using immunohistochemistry in a prostate cancer patient cohort (N=110). RESULTS: We found that conditioned media from LNCaP prostate cancer cells overexpressing Id-1 had a higher ability to drive osteoclast differentiation and a lower ability to stimulate osteoblast mineralisation than control, whereas conditioned media from PC3 prostate cancer cells with Id-1 knockdown were less able to stimulate osteoclast differentiation. Id-1 was found to negatively regulate TNF-beta and this correlation was confirmed in human prostate cancer specimens (P=0.03). Furthermore, addition of recombinant TNF-beta to LNCaP Id-1 cell-derived media blocked the effect of Id-1 overexpression on osteoblast mineralisation. CONCLUSION: In prostate cancer cells, the ability of Id-1 to modulate bone cell differentiation favouring metastatic bone disease is partially mediated by TNF beta, and Id-1 could be a potential therapeutic target for prostate cancer to bone metastasis. PMID- 20010942 TI - Epidermal growth factor signalling and bone metastasis. AB - Epidermal growth factor (EGF) signalling is well known for its multifaceted functions in development and tissue homoeostasis. The EGF family of ligands and receptors (ERBB family) have also been extensively investigated for their roles in promoting tumourigenesis and metastasis in a variety of cancer types. Recent findings indicate that EGF signalling is an important mediator of bone metastasis in breast, prostate and kidney cancers. The EGF signalling stimulates the growth of bone metastasis directly by increasing tumour cell proliferation and indirectly by engaging bone stromal cell in metastasis-promoting activities. Therefore, molecular targeting of ERBB receptors may benefit patients with bone metastasis and should be evaluated in clinical trials. PMID- 20010943 TI - Cisplatin-induced expression of Gb3 enables verotoxin-1 treatment of cisplatin resistance in malignant pleural mesothelioma cells. AB - BACKGROUND: A major problem with cisplatin treatment is the development of acquired-drug resistance of the tumour cells. Verotoxin-1 (VT-1) exerts its cytotoxicity by targeting the membrane glycolipid globotriasosylceramide (Gb3), a molecule associated with drug resistance. Cisplatin- and VT-1-induced apoptosis involves mitogen-activated protein kinase (MAPK) activation, and deactivation of MAPKs is associated with cisplatin resistance. This study aimed to investigate whether a sub-toxic concentration of VT-1 could enhance cisplatin-induced apoptosis and overcome acquired-cisplatin resistance in cultured cancer cell lines. METHOD: P31 and H1299 cells with corresponding cisplatin-resistant sub lines (P31res/H1299res) were incubated with VT-1 and/or cisplatin followed by determination of Gb3 expression, cell viability, apoptosis, and signalling pathways. RESULTS: Cells from the resistant sub-lines had elevated Gb3 expression compared with the parental cell lines, and cisplatin further increased Gb3 expression, whereas VT-1 reduced the percentage of Gb3-expressing cells. Combination of cisplatin and sub-toxic concentrations of VT-1 led to a super additive increase of cytotoxicity and TUNEL staining, especially in the cisplatin resistant sub-lines. Blockade of Gb3 synthesis by a Gb3 synthesis inhibitor not only led to eradicated TUNEL staining of P31 cells, but also sensitised P31res cells to the induction of apoptosis by cisplatin alone. Cisplatin- and VT-1 induced apoptosis involved the MAPK pathways with increased C-Jun N-terminal kinase and MAPK kinase-3 and -6 phosphorylation. CONCLUSIONS: We show the presence of Gb3 in acquired-cisplatin resistance in P31res and H1299res cells. Cisplatin up-regulated Gb3 expression in all cells and thus sensitised the cells to VT-1-induced cytotoxicity. A strong super-additive effect of combined cisplatin and a sub-toxic concentration of VT-1 in cisplatin-resistant malignant pleural mesothelioma cells were observed, indicating a new potential clinical treatment approach. PMID- 20010944 TI - Expression of aldehyde dehydrogenase 1 (ALDH1) is associated with basal-like markers and features of aggressive tumours in African breast cancer. AB - BACKGROUND: Putative breast cancer stem cells might express surface markers such as aldehyde dehydrogenase 1 (ALDH1) and BMI-1 proteins. The aim of this study was to explore the expression of these proteins in breast cancers from an African population and their associations with the basal-like phenotype (BLP) and other molecular characteristics. METHODS: We analysed 192 paraffin-embedded breast carcinoma samples by tissue microarrays and immunohistochemical methods. RESULTS: In total, 88 tumours (48%) expressed ALDH1, whereas 46 (25%) expressed BMI-1 protein. Expression of ALDH1 was associated with high histological grade (P<0.0005), high mitotic count (P<0.0005), high nuclear grade (P<0.0005), oestrogen receptor (ER) negativity (P<0.0005), progesterone receptor (PR) negativity (P=0.009), p53 expression (P=0.034), cytokeratin 5/6 positivity (P=0.008), epidermal growth factor receptor (EGFR) expression (P=0.015) and the BLP (P<0.0005), whereas it was inversely associated with BMI-1 staining (P=0.009). On the other hand, BMI-1 expression was associated with low histological grade (P=0.004) and ER positivity (P=0.001). CONCLUSION: There was a high prevalence of ALDH1 expression among breast carcinomas and associations with basal markers and features of aggressive tumours. Studies are required to elucidate the importance of these findings for improved understanding of breast cancer biology. PMID- 20010945 TI - Biomarkers of angiogenesis and their role in the development of VEGF inhibitors. AB - Vascular endothelial growth factor (VEGF) has been confirmed as an important therapeutic target in randomised clinical trials in multiple disease settings. However, the extent to which individual patients benefit from VEGF inhibitors is unclear. If we are to optimise the use of these drugs or develop combination regimens that build on this efficacy, it is critical to identify those patients who are likely to benefit, particularly as these agents can be toxic and are expensive. To this end, biomarkers have been evaluated in tissue, in circulation and by imaging. Consistent drug-induced increases in plasma VEGF-A and blood pressure, as well as reductions in soluble VEGF-R2 and dynamic contrast-enhanced MRI parameters have been reported. In some clinical trials, biomarker changes were statistically significant and associated with clinical end points, but there is considerable heterogeneity between studies that are to some extent attributable to methodological issues. On the basis of observations with these biomarkers, it is now appropriate to conduct detailed prospective studies to define a suite of predictive, pharmacodynamic and surrogate response biomarkers that identify those patients most likely to benefit from and monitor their response to this novel class of drugs. PMID- 20010946 TI - E-/P-selectins and colon carcinoma metastasis: first in vivo evidence for their crucial role in a clinically relevant model of spontaneous metastasis formation in the lung. AB - BACKGROUND: Interactions of endothelial selectins with tumour cell glycoconjugates have been shown to have a major role in tumour cell dissemination in previous experiments. However, experiments validating this observation were limited in value, as 'metastases' in these experiments were artificially induced by i.v. injection rather than developed spontaneously as in true metastases. METHODS: Endothelial (E) and platelet (P)-selectin-deficient severe combined immunodeficient (scid) mice were generated and human HT 29 colon cancer cells were subcutaneously inoculated in these mice and in wild-type scid mice. Tumour growth, spontaneous metastasis formation in the lung and adherence of HT29 cells to E- and P-selectin under flow were determined. RESULTS: The number of metastases decreased by 84% in E- and P-selectin-deficient scid mice, compared with wild-type scid mice. The remaining 16% metastases in the E- and P-selectin deficient scid mice grew within the pulmonary artery and not in the alveolar septae as they did in wild-type scid mice. Flow experiments indicate that tumour cells roll and tether on an E- and P-selectin matrix similar to leukocytes; however, firm adhesion is mainly mediated in E-selectin. CONCLUSION: Our results indicate that E- and P-selectins have a crucial role in spontaneous metastasis formation. As the human HT 29 colon cancer cells are positive for the lectin Helix pomatia agglutinin (HPA), which identified the metastatic phenotype in earlier clinical studies, these results are of particular clinical relevance. PMID- 20010947 TI - Body mass index and smoking-related lung cancer risk in the Singapore Chinese Health Study. AB - BACKGROUND: Smokers with low body mass index (BMI) may be more susceptible to lung cancer. METHODS: We prospectively examined the association between baseline BMI and lung cancer risk in the Singapore Chinese Health Study, a cohort of 63 257 Chinese enrolled between 1993 and 1998. RESULTS: After adjustment for smoking intensity and duration, BMI was inversely associated with risk of lung cancer among current smokers (P for trend=0.0004). Current smokers at different dosage of smoking with low BMI had significantly higher risk for lung cancer than those with high BMI. Hazard ratios (95% confidence intervals) of lung cancer for heavy smokers with BMI of > or =28, 24-<28, 20-<24, and <20 kg m(-2) were 6.37 (2.10 19.30), 9.01 (5.04-16.10), 8.53 (6.35-11.5), and 11.12 (6.60-18.70), respectively, as compared with nonsmokers. BMI had no modifying effects on lung cancer risk among nonsmokers and former smokers. CONCLUSION: Smokers with lower BMI may experience an enhanced risk of lung cancer. The findings have significant public-health implication given the increase in smoking prevalence in developing countries, where people still have relatively low BMI. PMID- 20010948 TI - CD133+ circulating haematopoietic progenitor cells predict for response to sorafenib plus erlotinib in non-small cell lung cancer patients. AB - BACKGROUND: Blood-based biomarkers may be particularly useful for patient selection and prediction of treatment response for angiogenesis inhibitors. Circulating endothelial cells (CECs) and haematopoietic progenitor cells (HPCs) might have a role in tumour angiogenesis and in tumour growth. Measurement of CECs and HPCs in the blood of patients could be a simple, non-invasive way to monitor or predict responses to treatment. METHODS: (VEGFR2(+)) CECs(,) (CD133(+)) HPCs, plasma vascular endothelial growth factor (VEGF) and erythropoietin were measured in blood from 25 non-small cell lung cancer (NSCLC) patients before and during treatment with sorafenib plus erlotinib (SO/ER). In order to assess the drug specificity of changes in CECs and HPCs, 18 patients treated with bevacizumab plus erlotinib (BV/ER) and 10 patients with erlotinib (ER) monotherapy were studied. Response was measured in all patient groups by Response Evaluation Criteria in Solid Tumors (RECIST). RESULTS: At day 7, SO/ER treated patients showed a three-fold increase in CECs (P<0.0001) comparable to BV/ER-treated patients (P<0.01), and the CECs did not change with erlotinib treatment (P=0.8). At day 7, CD133(+)/HPCs decreased with SO/ER treatment (P<0.0001). HPC numbers did not change with either BV/ER or erlotinib. In SO/ER treated patients pre-treatment CD133(+)/HPCs were significantly lower in responders (P=0.01) and pre-treatment CD133(+)/HPC numbers lower than the median correlated with a longer time-to-progression (TTP) (P=0.037). CONCLUSION: Pre treatment CD133(+)/HPCs are a promising candidate biomarker to further explore for use in selecting NSCLC patients who might benefit from SO/ER treatment. PMID- 20010950 TI - Emerging roles of Wnts in the adult nervous system. AB - The roles of the Wnt signalling pathway in several developmental processes, including synaptic differentiation, are well characterized. The expression of Wnt ligands and Wnt signalling components in the mature mammalian CNS suggests that this pathway might also play a part in synaptic maintenance and function. In fact, Wnts have a crucial role in synaptic physiology, as they modulate the synaptic vesicle cycle, the trafficking of neurotransmitter receptors and the interaction of these receptors with scaffold proteins in postsynaptic regions. In addition, Wnts participate in adult neurogenesis and protect excitatory synaptic terminals from amyloid-beta oligomer toxicity. Here, the latest insights into the function of Wnt signalling in the adult nervous system and therapeutic opportunities for neurodegenerative diseases such as Alzheimer's and Parkinson's disease are discussed. PMID- 20010949 TI - PSMB7 is associated with anthracycline resistance and is a prognostic biomarker in breast cancer. AB - BACKGROUND: To date individual markers have failed to correctly predict resistance against anticancer agents in breast cancer. We used gene expression patterns attributable to chemotherapy-resistant cells to detect potential new biomarkers related to anthracycline resistance. One of the genes, PSMB7, was selected for further functional studies and clinical validation. METHODS: We contrasted the expression profiles of four pairs of different human tumour cell lines and of their counterparts resistant to doxorubicin. Observed overexpression of PSMB7 in resistant cell lines was validated by immunohistochemistry. To examine its function in chemoresistance, we silenced the gene by RNA interference (RNAi) in doxorubicin-resistant MCF-7 breast cancer cells, then cell vitality was measured after doxorubicin treatment. Microarray gene expression from GEO raw microarray samples with available progression-free survival data was downloaded, and expression of PSMB7 was used for grouping samples. RESULTS: After doxorubicin treatment, 79.8+/-13.3% of resistant cells survived. Silencing of PSMB7 in resistant cells decreased survival to 31.8+/-6.4% (P>0.001). A similar effect was observed after paclitaxel treatment. In 1592 microarray samples, the patients with high PSMB7 expression had a significantly shorter survival than the patients with low expression (P<0.001). CONCLUSION: Our findings suggest that high PSMB7 expression is an unfavourable prognostic marker in breast cancer. PMID- 20010953 TI - Are correctional facilities amplifying the epidemic of community-acquired methicillin-resistant Staphylococcus aureus? PMID- 20010952 TI - Mass spectrometry tools for the classification and identification of bacteria. AB - Mass spectrometry has become an important analytical tool in biology in the past two decades. In principle, mass spectrometry offers high-throughput, sensitive and specific analysis for many applications in microbiology, including clinical diagnostics and environmental research. Recently, several mass spectrometry methods for the classification and identification of bacteria and other microorganisms, as well as new software analysis tools, have been developed. In this Review we discuss the application range of these mass spectrometry procedures and their potential for successful transfer into microbiology laboratories. PMID- 20010955 TI - Ischemic preconditioning regulates expression of microRNAs and a predicted target, MeCP2, in mouse cortex. AB - Preconditioning describes the ischemic stimulus that triggers an endogenous, neuroprotective response that protects the brain during a subsequent severe ischemic injury, a phenomenon known as 'tolerance'. Ischemic tolerance requires new protein synthesis, leads to genomic reprogramming of the brain's response to subsequent ischemia, and is transient. MicroRNAs (miRNAs) regulate posttranscriptional gene expression by exerting direct effects on messenger RNA (mRNA) translation. We examined miRNA expression in mouse cortex in response to preconditioning, ischemic injury, and tolerance. The results of our microarray analysis revealed that miRNA expression is consistently altered within each group, but that preconditioning was the foremost regulator of miRNAs. Our bioinformatic analysis results predicted that preconditioning-regulated miRNAs most prominently target mRNAs that encode transcriptional regulators; methyl-CpG binding protein 2 (MeCP2) was the most prominent target. No studies have linked MeCP2 to preconditioning or tolerance, yet miR-132, which regulates MeCP2 expression, is decreased in preconditioned cortex. Downregulation of miR-132 is consistent with our finding that preconditioning ischemia induces a rapid increase in MeCP2 protein, but not mRNA, in mouse cortex. These studies reveal that ischemic preconditioning regulates expression of miRNAs and their predicted targets in mouse brain cortex, and further suggest that miRNAs and MeCP2 could serve as effectors of ischemic preconditioning-induced tolerance. PMID- 20010956 TI - Neuronal estrogen receptor-alpha mediates neuroprotection by 17beta-estradiol. AB - 17beta-Estradiol (E(2)) was shown to exert neuroprotective effects both in in vitro and in vivo models of stroke. Although these effects of E(2) are known to require estrogen receptor-alpha (ER alpha), the cellular target of estrogen mediated neuroprotection remains unknown. Using cell type-specific ER mutant mice in an in vivo model of stroke, we specifically investigated the role of ER alpha in neuronal cells versus its role in the microglia in the mediation of neuroprotection by estrogens. We generated and analyzed two different tissue specific knockout mouse lines lacking ER alpha either in cells of myeloid lineage, including microglia, or in the neurons of the forebrain. Both E(2) treated and E(2)-untreated mutant and control mice were subjected to a permanent middle cerebral artery occlusion for 48 h, and the infarct volume was quantified. Although the infarct volume of E(2)-treated female myeloid-specific ER alpha knockout mice was similar to that of E(2)-treated control mice, both male and female neuron-specific ER alpha mutant mice had larger infarcts than did control mice after E(2) treatment. We conclude that neuronal ER alpha in female and male mice mediates neuroprotective estrogen effects in an in vivo mouse model of stroke, whereas microglial ER alpha is dispensable. PMID- 20010957 TI - Assessment of regional differences in tariquidar-induced P-glycoprotein modulation at the human blood-brain barrier. AB - We attempted to assess regional differences in cerebral P-glycoprotein (P-gp) function by performing paired positron emission tomography (PET) scans with the P gp substrate (R)-[(11)C]verapamil in five healthy subjects before and after i.v. infusion of tariquidar (2 mg/kg). Comparison of tariquidar-induced changes in distribution volumes (DVs) in 42 brain regions of interest (ROIs) failed to detect significant differences among brain ROIs. Statistical parametric mapping analysis of parametric DV images visualized symmetrical bilateral clusters with moderately higher DV increases in response to tariquidar administration in cerebellum, parahippocampal gyrus, olfactory gyrus, and middle temporal lobe and cortex, which might reflect moderately decreased P-gp function and expression. PMID- 20010959 TI - On the identification of human selected loci in grapevines. PMID- 20010960 TI - The clock gene cryptochrome of Bactrocera cucurbitae (Diptera: Tephritidae) in strains with different mating times. AB - Differences in mating time between populations can give rise to premating reproductive isolation. Tephritid fruit flies exhibit large variation in mating time among intra- or inter-specific populations. We previously cloned the clock gene period from two strains of melon fly, Bactrocera cucurbitae; in one the individuals mate early during the day, whereas in the other the individuals mate later. These strains were originally established by divergent artificial selection for developmental time, 'short' and 'long', with early and late mating times, respectively. The deduced amino acid sequences of PERIOD proteins for these two strains were reported to be identical. Here we cloned another clock gene cryptochrome (cry) from the two strains, and found two stable amino acid substitutions in the strains. In addition, the allele frequency at the two polymorphic sites of cry gene correlated with the circadian locomotor period (tau) across strains, whereas the expression pattern of cry mRNA in the heads of flies taken from the short strain significantly differed from that from the long strain. These findings suggest that variation in the cry gene is related to differences in the circadian behaviour in the two strains, thus implying that the cry gene may have an important role in reproductive isolation. PMID- 20010958 TI - Effects of AMP-activated protein kinase in cerebral ischemia. AB - AMP-activated protein kinase (AMPK) is a serine threonine kinase that is highly conserved through evolution. AMPK is found in most mammalian tissues including the brain. As a key metabolic and stress sensor/effector, AMPK is activated under conditions of nutrient deprivation, vigorous exercise, or heat shock. However, it is becoming increasingly recognized that changes in AMPK activation not only signal unmet metabolic needs, but also are involved in sensing and responding to 'cell stress', including ischemia. The downstream effect of AMPK activation is dependent on many factors, including the severity of the stressor as well as the tissue examined. This review discusses recent in vitro and in vivo studies performed in the brain/neuronal cells and vasculature that have contributed to our understanding of AMPK in stroke. Recent data on the potential role of AMPK in angiogenesis and neurogenesis and the interaction of AMPK with 3-hydroxy-3-methy glutaryl-CoA reductase inhibitors (statins) agents are highlighted. The interaction between AMPK and nitric oxide signaling is also discussed. PMID- 20010961 TI - Host-associated divergence and incipient speciation in the yucca moth Prodoxus coloradensis (Lepidoptera: Prodoxidae) on three species of host plants. AB - A wide range of evolutionary processes have been implicated in the diversification of yuccas and yucca moths, which exhibit ecological relationships that extend from obligate plant-pollinator mutualisms to commensalist herbivory. Prodoxus coloradensis (Lepidoptera: Prodoxidae) is a yucca moth, which feeds on the flowering stalks of three Yucca species as larvae, but does not provide pollination service. To test for evidence of host-associated speciation, we examined the genetic structure of P. coloradensis using mitochondrial (cytochrome oxidase I) and nuclear (elongation factor 1 alpha) DNA sequence data. Multilocus coalescent simulations indicate that moths on different host plant species are characterized by recent divergence and low levels of effective migration, with large effective population sizes and considerable retention of shared ancestral polymorphism. Although geographical distance explains a proportion of the mitochondrial and nuclear DNA variation among moths on different species of Yucca, the effect of host specificity on genetic distance remains significant after accounting for spatial isolation. The results of this study indicate that differentiation within P. coloradensis is consistent with the evolution of incipient species affiliated with different host plants, potentially influenced by sex-biased dispersal and female philopatry. PMID- 20010962 TI - Diabetes: intensive glucose-lowering therapy-weighing up the evidence. PMID- 20010963 TI - Bone: causes of low bone mass in breast cancer-time for action? PMID- 20010964 TI - Pediatric endocrinology: vitamin D and cardiovascular disease risk in children. PMID- 20010965 TI - Growth and development: congenital adrenal hyperplasia-glucocorticoids and height. PMID- 20010966 TI - Parathyroid gland: cinacalcet-can it control hypercalcemia? PMID- 20010967 TI - Bone: bone density screening leads to reduced fracture risk. PMID- 20010968 TI - Hereditary hemochromatosis and diabetes mellitus: implications for clinical practice. AB - Hereditary hemochromatosis (HH) is a genetic condition that can lead to unregulated absorption of iron from the gut with resultant iron overload. The most common form of HH is caused by mutations in the HFE gene, with most cases of HH presenting in patients who are homozygous for the Cys282Tyr mutation. The prevalence of HFE gene mutations in persons of Northern European ancestry is fairly high (0.3-0.7% homozygous and 9-14% heterozygous for the Cys282Tyr mutation), but the penetrance of the disease is considered fairly low and is quite variable. While routine screening of the general population is not recommended, a targeted approach to screening in symptomatic patients and in those with a family member with iron overload is warranted. Untreated, iron overload can lead to considerable morbidity including liver cirrhosis, arthritis and diabetes mellitus, and increased mortality. The pathophysiology of diabetes mellitus in HH is thought to be due primarily to defects in the early insulin response to glucose. An Hfe(-/-) mouse model of HH has demonstrated defects in beta-cell function and beta-cell apoptosis that may be mediated by increased oxidative stress. Fortunately, these defects seem to be reversible if phlebotomy treatment is initiated before the development of cirrhosis or diabetes mellitus in patients. Further research into the long-term effects of treatment on prevention of diabetes mellitus in HH is needed. PMID- 20010969 TI - Management of osteomyelitis of the foot in diabetes mellitus. AB - Although osteomyelitis occurs in approximately 10-20% of patients with diabetes related foot ulcers, no widely accepted guideline is available for its treatment. In particular, little consensus exists on the place of surgery. A number of experts claim that early surgical excision of all infected or necrotic bone is essential. Others suggest that surgery should not be performed routinely, but instead only in patients who do not respond to antibiotic treatment or in case of particular clinical indications. Unfortunately, no studies have directly compared the two approaches. Over 500 cases of conservative (that is, nonsurgical) management with resolution rates of 60-80% have been described previously. Most patients in these series, however, received prolonged courses of broad-spectrum antibiotics, which increase the risk of diarrhea caused by Clostridium difficile or the emergence of multidrug-resistant organisms. By contrast, relatively few series of primarily surgical management have been published, with widely differing outcomes, and some of them also reported high recurrence rates. Further research is required to establish the relative importance of each approach, but the available data clearly indicate that a combined assessment and treatment by surgeons and physicians together is essential for many patients. PMID- 20010970 TI - Harvey Cushing's attempt at the first human pituitary transplantation. AB - BACKGROUND: This case study illustrates Harvey Cushing's pioneering work in pituitary transplantation in the early 20th century and the essential relationship between laboratory research and clinical practice. In 1911, a 48 year-old man presented at Johns Hopkins Hospital with bitemporal hemianopsia, hypothermia, hypersomnolence, decreased libido, polydypsia and polyuria. INVESTIGATION: A review of the Johns Hopkins Hospital surgical records from 1896 1912 on a patient with hypopituitarism secondary to a suprasellar mass, in whom the first documented pituitary gland transplantation was performed. DIAGNOSIS: A diagnosis of hypopituitarism was made. Postmortem examination revealed a cystic cavity lined with squamous epithelium. MANAGEMENT: The patient was treated with whole-gland pituitary extract, which improved his symptoms only temporarily. Cushing transplanted a pituitary gland obtained from a spontaneously aborted fetus into the cerebral cortex of the patient, who showed marked improvement of his somnolence and confusion, whereas his polyuria and polydypsia persisted. A recurrence of symptoms after 6 weeks prompted Cushing to attempt a second transplant of a fetal pituitary gland, without improvement. The patient resumed hormonal supplementation with whole-gland pituitary extract, but died a month after the second transplant from respiratory complications. PMID- 20010971 TI - Making claims: functional foods for managing appetite and weight. AB - Functional food products promote claims such as 'freedom from hunger' and 'feel fuller for longer'. A legislative framework has been established by the European Food Safety Authority to evaluate the validity of such claims: a claim must be substantiated by scientific evidence and should be clearly understood by consumers. Since consumed foods influence appetite by means of a system of physiological satiety signals, functional foods could in principle act by increasing the potency and/or duration of these signals. Importantly, what constitutes a useful action: a reduction in hunger, an increase in fullness, a change in food intake at a meal, an adjustment in daily energy balance or a reduction in body weight? Any claim should not go beyond the scientific evidence of an effect, and methods exist to scientifically evaluate claims. The wording of a claim is, therefore, critical. The difference between a proof of concept and a guarantee of success is an important point that needs to be conveyed to the consumer. PMID- 20010972 TI - Dendritic tiling through TOR signalling. PMID- 20010974 TI - De Barsy syndrome and ATP6V0A2-CDG. PMID- 20010975 TI - Internal exposure to pollutants and sex hormone levels in Flemish male adolescents in a cross-sectional study: associations and dose-response relationships. AB - Flanders is densely populated with much industry and intensive farming. Hormonal status of 14- to 15-year-old male adolescents was studied in relation to internal exposure to pollutants. A total of 887 participants were selected as a random sample of the adolescents residing in the study areas. Confounding factors and significant covariates were taken into account. Serum levels of testosterone, free testosterone and estradiol, and the aromatase index showed significant positive associations with serum levels of marker polychlorobiphenyls (sum of PCBs 138, 153, and 180) and of hexachlorobenzene (HCB) and a negative association with urinary cadmium concentration. Serum levels of estradiol also showed a positive association with serum levels of dichlorodiphenyldichloroethylene (DDE). A doubling of serum concentrations of marker PCBs and HCB and of urinary concentration of cadmium were, respectively, associated with an increase of 16.4% (P<0.00001) and 16.6% (P<0.001) and a decrease of 9.6% (P<0.001) in serum testosterone concentration. Similar findings were made after additional adjustment for concurrent exposures. Associations between biological effects and internal exposures were, in terms of the regression coefficient, often stronger at exposures below the median. Environmental exposures to pollutants resulting in "normal" levels of internal exposure were associated with quite substantial differences in hormone concentrations. PMID- 20010976 TI - Selection bias in case-control studies on household exposure to pesticides and childhood acute leukemia. AB - The goal of this study was to investigate the potential for selection bias in published case-control studies on household exposure to pesticides and childhood acute leukemia; most studies have reported positive findings. Items to evaluate the potential for selection bias were first developed. They focused on the source populations that gave rise to cases and controls, the probabilistic selection of subjects from the source, and the losses of the subjects actually selected. A quantitative assessment of bias was also carried out. Potential sources of selection bias were found in all the studies, but none of them were observed across all the studies. Main sources of potential bias were a non-concurrent selection of controls with respect to cases, the use of control diagnoses possibly caused by pesticide exposure in hospital-based studies, and non participation of selected eligible subjects. A quantitative assessment of bias concluded that non-participation alone could not explain the reported positive associations. We conclude that overall, selection bias, as a likely source of bias in these studies, does not seem to explain their positive findings. Our analysis provides arguments strengthening the conclusions on associations reported in earlier studies. PMID- 20010977 TI - Estimated daily intake of phthalates in occupationally exposed groups. AB - Improved analytical methods for measuring urinary phthalate metabolites have resulted in biomarker-based estimates of phthalate daily intake for the general population, but not for occupationally exposed groups. In 2003-2005, we recruited 156 workers from eight industries where materials containing diethyl phthalate (DEP), dibutyl phthalate (DBP), and/or di(2-ethylhexyl) phthalate (DEHP) were used as part of the worker's regular job duties. Phthalate metabolite concentrations measured in the workers' end-shift urine samples were used in a simple pharmacokinetic model to estimate phthalate daily intake. DEHP intake estimates based on three DEHP metabolites combined were 0.6-850 MUg/kg/day, with the two highest geometric mean (GM) intakes in polyvinyl chloride (PVC) film manufacturing (17 MUg/kg/day) and PVC compounding (12 MUg/kg/day). All industries, except phthalate manufacturing, had some workers whose DEHP exposure exceeded the U.S. reference dose (RfD) of 20 MUg/kg/day. A few workers also exceeded the DEHP European tolerable daily intake (TDI) of 50 MUg/kg/day. DEP intake estimates were 0.5-170 MUg/kg/day, with the highest GM in phthalate manufacturing (27 MUg/kg/day). DBP intake estimates were 0.1-76 MUg/kg/day, with the highest GMs in rubber gasket and in phthalate manufacturing (17 MUg/kg/day, each). No DEP or DBP intake estimates exceeded their respective RfDs. The DBP TDI (10 MUg/kg/day) was exceeded in three rubber industries and in phthalate manufacturing. These intake estimates are subject to several uncertainties; however, an occupational contribution to phthalate daily intake is clearly indicated in some industries. PMID- 20010978 TI - Infants' exposure to aluminum from vaccines and breast milk during the first 6 months. AB - The success of vaccination programs in reducing and eliminating infectious diseases has contributed to an ever-increasing number of vaccines given at earlier ages (newborns and infants). Exposure to low levels of environmental toxic substances (including metals) at an early age raises plausible concerns over increasingly lower neuro-cognitive rates. Current immunization schedules with vaccines containing aluminum (as adjuvant) are given to infants, but thimerosal (as preservative) is found mostly in vaccines used in non industrialized countries. Exclusively, breastfed infants (in Brazil) receiving a full recommended schedule of immunizations showed an exceedingly high exposure of Al (225 to 1750 MUg per dose) when compared with estimated levels absorbed from breast milk (2.0 MUg). This study does not dispute the safety of vaccines but reinforces the need to study long-term effects of early exposure to neuro-toxic substances on the developing brain. Pragmatic vaccine safety needs to embrace conventional toxicology, addressing especial characteristics of unborn fetuses, neonates and infants exposed to low levels of aluminum, and ethylmercury traditionally considered innocuous to the central nervous system. PMID- 20010979 TI - [Cultural transition and public health]. PMID- 20010980 TI - [Alcohol habits and acute alcohol intoxication among middle school students]. AB - The objective of this study was to evaluate the age at which alcohol consumption begins and assess alcohol habits among young adolescents, in order to implement and evaluate specific preventive measures aimed at reducing the impact of alcohol related conditions. Students of a middle school in the province of Catania (Italy) were interviewed anonymously, through a self-administered semi-structured questionnaire. Overall, 432 students aged 10-14 years (47% male and 53% female) participated in the study. Results show that 52% of students have tried alcoholic drinks, and that alcohol consumption is more frequent among males with respect to females (54.2% versus 41.9%; p = 0.03). and increases with increasing year attended at school (1st year: 43.2%; 2nd year: 44.1%; 3rd year: 55.3%, Chi-square for trend p = 0.03). The average alcohol intake among drinkers is 7.0 g/day. Fourteen percent of students are regular drinkers and a higher percentage of regular drinkers can be found among the male population compared to the female population (19.7% versus 8.7% respectively; p = 0.001). Finally, 8% of participating students admitted having been drunk at least once in their lifetime. PMID- 20010981 TI - [Surveillance of nosocomial infections in a cardiac surgery unit in Palermo (Italy)]. AB - A study was performed to assess the incidence of hospital acquired infections at the Cardiac Surgey/Intensive Care Unit of the university teaching hospital "Paolo Gaccione" in Palermo (Italy), their associated risk factors and mortality. The unit consists of 31 beds, including 7 intensive care beds. One hundred patients admitted to the unit between 1 March 2008 and 30 June 2008 and hospitalised for > 48 hours were enrolled in the study. Active surveillance of hospital acquired infections was performed for these patients and overall, 18 cases of infection were diagnosed (9.3 infections per 1.000 day-patient). Infections acquired in hospital were found to be associated with several risk factors (duration of stay, duration of the surgical procedure and number of persons in the operating theatre during the procedure) and with a higher mortality rate (OR=5.9). Knowledge of the factors associated with hospital infections is essential to reducing human and social costs related to such infections while simultaneously improving the quality of health care. PMID- 20010982 TI - Health authority: the Italian experience. AB - The healthcare reform, introduced in Italy in 1992, has completely changed the structure of the national healthcare system (NHS) , including the introduction of the concept of "business firm" applied to public health service providers. The aim of this study was to outline the history of healthcare "firms" (azienda sanitaria) and evaluate the impact of this change on the NHS in terms of health expenditure, and corporate effectiveness and efficiency. Self regulation and correction are the abilities to which the success of healthcare companies can be attributed. The benefits of creating healthcare firms include preventing those problems associated with healthcare models based on the principles of the private insurance type model and preferring instead a cost-effectiveness approach. PMID- 20010983 TI - [Multiple chemical sensitivity: a study in a young adult population in Italy]. AB - Multiple chemical sensitivity (MCS) is a chronic recurrent condition characterized by reduced tolerance to various environmental agents or to a class of chemical substances. An evidence-based diagnostic panel is not available nor is there a universally accepted definition of the syndrome. This study retrospectively evaluated the prevalence of MCS symptoms and associated risk factors in a sample of 660 young adults, by means of a structured questionnaire. Symptoms of MCS were found to be widely distributed in the sample population and several of the main risk factors suspected to be associated with MCS were not consistently associated with these symptoms in the sample. On the other hand a significant correlation (p<0.05) was found between atopy and symptoms. Further studies are therefore needed to evaluate this association and reach a unifying definition of the syndrome. PMID- 20010984 TI - [Critical aspects of the direct distribution of medicines to patients upon discharge at the G. Rummo hospital in Benevento (Italy)]. AB - Delivering prescription drugs directly to patients at the moment of discharge from the hospital is a useful tool for ensuring continuity in patient drug use, improving prescriptive appropriateness, limiting pharmaceutical expenditure and analyzing pharmacoepidemiological data. A project was therefore conducted from April 2005 to January 2007, at the G. Rumino hospital in Benevento (Italy), to encourage the direct delivery of drugs to patients upon discharge. The project consisted of various phases. Firstly, the medical records of all patients discharged from the hospital during April 2005 were analysed, mainly to collect information regarding discharge prescriptions, verify whether copies of the discharge form and prescription records were present in the chart, the type of drugs prescribed and whether these were available in the hospital pharmacy list of available drugs and had been dispensed to the patient. The percentage of drugs not available and of patients who did not pick up the prescribed drugs was calculated, critical aspects of the prescription process were analysed, and corrective measures implemented. A second evaluation of medical records was then performed for patients discharged in January 2007, to evaluate the effectiveness of the corrective measures applied. Results show that most discharged patients continue not to take advantage of the direct distribution of drugs in hospital and more information and communication to physicians and patients regarding this opportunity is required. PMID- 20010985 TI - [Methods of conservation, safety and nutritional quality of industrial originating food]. PMID- 20010986 TI - [Medicine and physical activity and their relatedness in history]. AB - The authors outline how scientific and social ideas and concepts, regarding the role of physical activity in human health, have evolved throughout the last centuries,. PMID- 20010987 TI - [Hepatitis A in Europe today: epidemiology and prevention]. AB - The susceptibility to Hepatitis A virus infection is growing among the European population and many hepatitis outbreaks have been described in 2004-2008. The change in epidemiological characteristics observed in the last decades has to be taken in account in issuing specific recommendations about the use of the vaccine in pre and post exposure prophylaxis. PMID- 20010988 TI - [Public health research and proceedings of the XI Public Health Conference]. PMID- 20010989 TI - [Drug resistance in endemic and imported tuberculosis cases in Sicily (Italy)]. AB - This study was performed to evaluate the prevalence of anti-tuberculosis (TB) drug resistance in Sicily (Italy). A total of 92 Mycobacterium tuberculosis strains were tested, 43 of which were isolated from outpatients between 2004 and 2007 and 49 isolated from patients hospitalized in various eastern Sicilian hospitals. The 92 strains were all isolated from adult patients (mean age 56 years), 71 of which were Italian and the remaining 21non-Italian. Drug sensitivity testing was performed by using the Canetti proportion method and revealed a large proportion of strains (48.9%) found to be resistant to one or more drugs. The most frequent type of resistance was found to be towards rifampicin (38%). Simultaneous resistance to rifampicin and isoniazid, which characterizes multi-drug resistant TB (MDR-TB), was observed in 19.5% of strains; this was always associated with resistance to at least one other anti tuberculosis drug. No significant differences were found in the distribution of drug resistance, including multi-drug resistance, between strains isolated from Italian and non-Italian patients. A higher percentage of resistance was found amongst strains isolated from hospitalized patients with respect to those from outpatients: rifampicin 53% vs 21%; isoniazid 41% vs 9%; Mdr-TB 47% vs 12%. PMID- 20010990 TI - [A study to compare formal duties of medical directors of Italian public hospitals with actual activities performer]. AB - It is difficult to identify, from a regulatory point of view, the actual competencies of medical directors of public hospitals in Italy. A questionnaire survey was therefore conducted to identify which are the activities actually performed by this professional. One hundred twenty local health authorities participated in the survey. Findings reveal that many activities conducted by the hospital medical director, such a mediation with the local health authorities, are not formally recognized and that the medical director actually conducts more complex activities than those formally pertaining to his role. PMID- 20010991 TI - [Quality evaluation of informed consent forms: a pilot study]. AB - Informed consent forms must be designed to meet specific requirements but evaluating the structure and content requirements of consent forms from a qualitative point of view is not a simple task. An evaluation tool was therefore developed in the context of a pilot study to evaluate the current quality of informed consent forms at both medical and surgical departments of the San Giovanni Battista Hospital in Turin (Italy). A random sample of 288 medical records of patients hospitalized during 2007-2008 was selected. These were evaluated to determine the presence of a properly executed informed consent form or of a justification for its absence. Each consent form was evaluated based on the presence of a list of requirements. Statistical analysis was conducted by first taking into account all departments and then stratifying by medical and surgical departments. The evaluation tool was found to useful in evaluating, in a standardized and repeatable manner, the adequacy and completeness of informed consent forms in different hospital departments. PMID- 20010993 TI - [Surveillance of the health status of the population of the city of Parona (Italy), Phase 2]. AB - This study is part of a prospective study, which started in the year 2000, to monitor the health status of the population of the city of Parona, a previously agricultural area which has been transformed in a prevalently industrial area. This is the second phase of a project regarding the epidemiological surveillance of this population. The aim of the project was to detect changes in the health status of the population, particularly with regard to respiratory diseases associated with air pollution. Data was collected through the use of a questionnaire and a spirometry test. A control group was used in both parts of the study; control subjects lived in the city of Rovescala. In total 1484 subjects were invited by letter to participate and 399 (27%) accepted. The mean age of participants was 48 years (range 15-74 years). Only 149/399 (37%) subjects had participated in part 1 of the study. No significant changes were found in respiratory symptoms during the period considered. Three hundred eighty six subjects underwent spirometry testing, 36 of which (9%) were found to have an abnormal result. A moderately increased prevalence of obstructive airway signs on spirometry was registered in female participants, with respect to the year 2000 (5.9% vs 2.6%). On the other hand, the prevalence of restrictive signs remained similar in both males (1,7% vs 2,4%) and females (2% vs 1,5%). The spirometry changes detected in the149 subjects (65 male and 84 female) who participated in both parts of the study (parts 1 and 2) were most likely age-related. No significant differences were found with respect to the control group, although the prevalence of obstructive signs among males was greater in the city of Parona (9.2% vs 6.9%. PMID- 20010992 TI - [Epidemiology of tumour mortality from 2000 to 2005 in the province of Vercelli (Italy)]. AB - The main aim of this study was to assess the burden of cancer disease in residents of Vercelli (northern Italy), by analysing mortality data and hospital discharge forms. This was done in order to verify whether, as widely suspected among the city's population, an increased risk of cancer exists in the area, due to the large number of existing agricultural and industrial activities. Tumour mortality rates were compared with mortality data from the tumour registries of the province of Biella and of the city of Turin to identify a possible excess number of cases in Vercelli.An increased mortality rate was observed with respect to reference values (mortality registry of the city of Turin) for several tumours; more specifically results revealed a significantly increased mortality rate due to colorectal tumours, leukemias and nervous system tumours in both genders. Excess mortality was detected in males but not in females for esophageal cancer, non melanoma skin cancers, pancreatic, laryngeal, prostatic, renal and bone cancers. Conversely, in females, the standard mortality ratios (SMR) were found to be statistically significant for renal and laryngeal cancers and for Hodgkin's lymphoma. PMID- 20010994 TI - [Cutaneous leishmaniasis in Catania (Italy): persistently endemic and underestimated]. AB - Cases of cutaneous leishmaniasis diagnosed at the laboratory of Department of Hygiene of the University of Catania (Italy), from 1998-2007 were evaluated. Specifically, demographic and personal data, area of origin and temporal trend of the 97 cases confirmed by microscopy in this 10-year period were examined. The data collected shows that the disease is still widely present in this area ; on the other hand official data regarding this disease appears incomplete and shows various inconsistencies. This is due to underreporting but also to scarce knowledge of the disease by physicians. Elimination of cutaneous leishmaniasis can be achieved by controlling leishmaniasis in dogs and improving environmental conditions; this requires greater efforts not only by institutions but also by individuals. PMID- 20010995 TI - [A survey to evaluate and compare public health research in Northern, Southern and Eastern Europe]. AB - A study was performed to describe public health research in Northern, Southern and Eastern Europe. A questionnaire was developed in collaboration with SPHERE (Strengthening Public Health Research in Europe) and the national public health associations of 23 countries participated in the Survey. A common problem amongst participating countries, that emerged from survey results, was found to be the lack of infrastructure and personnel; differences in future priorities were found. There is a need to strengthen collaboration between public health associations and policy makers in order to find effective solutions to common problems. PMID- 20010996 TI - [World Health Organization Guidelines for swimming pools and similar recreational water environments: Italian translation by the SItI Working Group "Enhancing health and physical activity"]. AB - The Working Group "Enhancing Health and Physical Activity" of the Italian Society of Hygiene, Preventive Medicine and Public Health promoted the Italian translation of the WHO Guidelines for safe recreational waters, swimming pools and similar environments. The structure of the WHO document is described and the main chapters are summarised. References are included from Italian studies and data provided by the Working Group. PMID- 20010997 TI - [Use of the new Italian version of the International Classification of Diseases Clinical Modification (ICD-9-CM) for the information system of immunization health services]. AB - Compared to the previous one, the recently updated Italian version of the International Classification of Diseases - Clinical Modification (ICD-9-CM) includes new codes for the description of medical-related vaccinations procedures. In this work the new codes are described and the Classification is proposed as a possible tool for storing electronic information systems of immunization health services. PMID- 20010998 TI - Anterior corneal profile with variable asphericity. AB - We present a corneal profile in which the eccentricity, e(Q=-e(2)), has a nonlinear continuous variation from the center outwards. This nonlinear variation is intended to fit and reproduce our current experimental data in which the anterior corneal surface of the human eye exhibits different values of e at different diameters. According to our clinical data, the variation is similar to an exponential decay. We propose a linear combination of two exponential functions to describe the variation of e. We then calculate the corneal sagittal height by substituting e in the first-order aspherical surface equation to obtain the corneal profile. This corneal profile will be used as a reference to analyze the resultant profiles of the customized corneal ablation in refractive surgery. PMID- 20010999 TI - Design and analysis of a surface plasmon polariton modulator using the electro optic effect. AB - A metal-insulator-metal (MIM) waveguide with a periodic change of dielectric materials in its insulator layer is proposed to create a voltage-controlled one dimensional plasmonic Bragg reflector. KNbO(3) is used as one portion of the dielectric material so that the refractive index of the insulator layer can be varied by an external applied voltage. In our analysis, the transfer matrix method is employed to design and optimize the transmission spectra of the MIM waveguide Bragg reflector. The reflectivity at the on (bias voltage of 65 V) and off (bias voltage of -65 V) states is shown to be >74% and <7%, respectively, for the operating wavelength of 1.5 mum. The results are also verified by the finite difference time-domain method. Furthermore, the modulation response of the MIM waveguide is studied. For the device area and average insulator layer thickness to be 10 mumx40 mum and 615 nm, respectively, it is found that the turn-on reflectivity of the waveguide is only slightly reduced to 69% at the -3 dB point in comparison with 75% for its steady-state performance. PMID- 20011000 TI - Robust wafer identification recognition based on asterisk-shape filter and high low score comparison method. AB - Wafer identifications (wafer ID) can be used to identify wafers from each other so that wafer processing can be traced easily. Wafer ID recognition is one of the problems of optical character recognition. The process to recognize wafer IDs is similar to that used in recognizing car license-plate characters. However, due to some unique characteristics, such as the irregular space between two characters and the unsuccessive strokes of wafer ID, it will not get a good result to recognize wafer ID by directly utilizing the approaches used in car license-plate character recognition. Wafer ID scratches are engraved by a laser scribe almost along the following four fixed directions: horizontal, vertical, plus 45 degrees , and minus 45 degrees orientations. The closer to the center line of a wafer ID scratch, the higher the gray level will be. These and other characteristics increase the difficulty to recognize the wafer ID. In this paper a wafer ID recognition scheme based on an asterisk-shape filter and a high-low score comparison method is proposed to cope with the serious influence of uneven luminance and make recognition more efficiently. Our proposed approach consists of some processing stages. Especially in the final recognition stage, a template matching method combined with stroke analysis is used as a recognizing scheme. This is because wafer IDs are composed of Semiconductor Equipment and Materials International (SEMI) standard Arabic numbers and English alphabets, and thus the template ID images are easy to obtain. Furthermore, compared with the approach that requires prior training, such as a support vector machine, which often needs a large amount of training image samples, no prior training is required for our approach. The testing results show that our proposed scheme can efficiently and correctly segment out and recognize the wafer ID with high performance. PMID- 20011001 TI - Arbitrary shaped microlens array laser sintered on glass sheets. AB - A new technique based on laser vitrification of cordierite ceramic powders is used to fabricate arbitrarily shaped microlens arrays on a glass substrate. Crack free semispherical and quasi-spherical lenses with good optical and surface quality are demonstrated. PMID- 20011002 TI - Freeform surface lens for LED uniform illumination. AB - A method using a freeform surface lens for LED secondary optic design is proposed in this paper. By Snell's Law, the differential equations are given to build the relationship between the normal direction of a freeform surface and its input/output ray vectors. Runge-Kutta formulas are used to calculate the differential equations to design the freeform surface. Moreover, the optical model for uniform illumination is simulated and optical performance is analyzed. A practical freeform surface lens for LED uniform illumination is fabricated using an injection molding method. By the process, our system demonstrates a uniform illumination with a divergence half-angle of 6 degrees and an efficiency of 78.6%. PMID- 20011003 TI - New artificial dielectric metamaterial and its application as a terahertz antireflection coating. AB - We describe a novel artificial dielectric material that has applications at millimeter and submillimeter wavelengths. The material is manufactured from layers of metal mesh patterned onto thin polypropylene sheets, which are then bonded together using a hot pressing process to provide planar rugged discs that can be reliably cycled to cryogenic temperatures. The refractive index of this material can be tuned by adjusting the geometry and spacing of the metal mesh layers. We demonstrate its usage by designing and characterizing a broadband antireflection coating for a Z-cut crystalline quartz plate. The coating was fabricated and applied to the quartz using the hot press technique and characterized using a Fourier transform spectrometer. The performance is shown to be in good agreement with a high frequency structure simulator and transmission line modeling results. PMID- 20011004 TI - Simultaneous laser-induced fluorescence and Raman imaging inside a hydrogen engine. AB - We report on the simultaneous and two-dimensional measurement of laser-induced fluorescence (LIF) and Raman scattering (Ramanography) applied inside a hydrogen internal combustion (IC) engine. Two different LIF tracer molecules, triethylamine (TEA) and trimethylamine (TMA), were used for the LIF experiments. The LIF and Raman results were found to be in very good agreement. The simultaneous application of Ramanography and LIF imaging indicated that TMA is the more suitable LIF tracer molecule, compared to TEA. PMID- 20011005 TI - Polarization induced instabilities in external four-mirror Fabry-Perot cavities. AB - Various four-mirror optical resonators are studied from the perspective of realizing passive stacking cavities. A comparative study of the mechanical stability is provided. The polarization properties of the cavity eigenmodes are described, and it is shown that the effect of mirror misalignments (or motions) induces polarization and stacking power instabilities. These instabilities increase with the finesse of the Fabry-Perot cavity. A tetrahedral configuration of the four mirrors is found to minimize the consequences of the mirrors' motion and misalignment by reducing the instability parameter by at least 2 orders of magnitude. PMID- 20011006 TI - Layered silver nanoparticles embedded in a BaF(2) matrix: optical characterization. AB - Multilayer stacks of silver and BaF(2) alternate layers have been deposited by thermal evaporation on a silica substrate with the aim to obtain Ag clusters dispersed in a BaF(2) insulator matrix. The Ag layer thickness was approximately 1.2 nm; the thickness of the BaF(2) layer was approximately 25 nm. The samples were thermally treated for a 1 h thermal annealing process at 500 degrees C. These kinds of multilayer device also have several applications in the field of optics for the realization of antireflection coatings. However, optical characterization of dielectric matrices that contain layered metallic nanoparticles still remains an unsolved problem in the field of nanostructured optical coatings. Therefore, the surface plasmon resonance peak that appears in the optical absorption spectra because of the formation of Ag nanoclusters inside the BaF(2) insulator matrix has been monitored and fitted by numerical codes. In particular, a previously published theoretical model, based on the Maxwell Garnett effective medium theory, modified to take into account the effects that are due to the particle shapes and the spatial arrangement of the clusters, has been employed to fit the optical absorption spectra. PMID- 20011007 TI - Simple and accurate expressions for diffuse reflectance of semi-infinite and two layer absorbing and scattering media. AB - We present computationally efficient and accurate semiempirical models of light transfer suitable for real-time diffuse reflectance spectroscopy. The models predict the diffuse reflectance of both (i) semi-infinite homogeneous and (ii) two-layer media exposed to normal and collimated light. The two-layer medium consisted of a plane-parallel slab of finite thickness over a semi-infinite layer with identical index of refraction but different absorption and scattering properties. The model accounted for absorption and anisotropic scattering, as well as for internal reflection at the medium/air interface. All media were assumed to be nonemitting, strongly forward scattering, with indices of refraction between 1.00 and 1.44 and transport single-scattering albedos between 0.50 and 0.99. First, simple analytical expressions for the diffuse reflectance of the semi-infinite and two-layer media considered were derived using the two flux approximation. Then, parameters appearing in the analytical expression previously derived were instead fitted to match results from more accurate Monte Carlo simulations. A single semiempirical parameter was sufficient to relate the diffuse reflectance to the radiative properties and thickness of the semi infinite and two-layer media. The present model can be used for a wide range of applications including noninvasive diagnosis of biological tissue. PMID- 20011008 TI - Optical properties of boron carbide near the boron K edge evaluated by soft-x-ray reflectometry from a Ru/B(4)C multilayer. AB - Soft-x-ray Bragg reflection from two Ru/B(4)C multilayers with 10 and 63 periods was used for independent determination of both real and imaginary parts of the refractive index n = 1 - delta + ibeta close to the boron K edge (approximately 188 eV). Prior to soft x-ray measurements, the structural parameters of the multilayers were determined by x-ray reflectometry using hard x rays. For the 63 period sample, the optical properties based on the predictions made for elemental boron major deviations were found close to the K edge of boron for the 10-period sample explained by chemical bonding of boron to B(4)C and various boron oxides. PMID- 20011009 TI - Mode stability of external cavity diode lasers. AB - Mode stability is an important performance characteristic of external cavity diode lasers (ECDLs). It has been well established that the continuous mode-hop free tuning range of a grating-feedback ECDL can be optimized by rotating the grating about a specific pivot location. We show that similar results can be obtained for other more convenient pivot locations by choosing instead the cavity length and grating location. The relative importance of the temperature stability of the diode and of the external cavity is also evaluated. We show that mechanically simple ECDL designs, using mostly standard components, can readily achieve a 35 GHz mode-hop-free tuning range at 780 nm. PMID- 20011010 TI - Semianalytical thermal analysis of the heat capacity of YAG laser rods. AB - Based on the theory of semianalytical thermal analysis, we investigate the heat capacity of Nd:YAG laser rods for pumping and cooling. A general expression for the temperature field within Nd:YAG laser rod crystals is obtained for the pumping stage and the relation of the maximum temperature rise with pumping time. We also achieve an expression for the temperature field for the cooling stage and the relation of the maximum temperature rise with cooling time. These results show that, when using the output power of 300 W LD pumped Nd:YAG laser rod crystals for 5 s, the maximum temperature rise in the center of the pump face is 154.79 degrees C. After we stop the pumping for 30 s, the maximum temperature rise drops to 0.8%. These results are in agreement with those reported by others. Our results provide a theoretical basis for the optimized design of a LD end pumped heat capacity laser. PMID- 20011011 TI - Optical coherence tomography for quantitative surface topography. AB - We address the measurement and correction of fan distortion in optical coherence tomography (OCT). This effect arises from the scanning system configuration and prevents one, in general, from obtaining quantitative topographic data from OCT. Computer simulations allowed us to quantify the effect and evaluate its dependence on the scanning mirror separation and design of the collimating lens, as well as to estimate the optimal axial position of that lens to minimize the fan distortion. We also developed a numerical algorithm based on 3-D ray propagation for the correction of the residual fan distortion. The effect was studied experimentally using a custom developed time-domain OCT in a Michelson configuration provided with a confocal channel, and the accuracy of the fan distortion correction algorithm tested on samples of known dimensions (flat surfaces and spherical lenses). With a proper calibration of the system with use of an onfocal channel, this algorithm makes it possible for time-domain OCT devices to be used as topographers. PMID- 20011012 TI - Thermal and near infrared sensor for carbon observation Fourier-transform spectrometer on the Greenhouse Gases Observing Satellite for greenhouse gases monitoring. AB - The Greenhouse Gases Observing Satellite (GOSAT) monitors carbon dioxide (CO(2)) and methane (CH(4)) globally from space using two instruments. The Thermal and Near Infrared Sensor for Carbon Observation Fourier-Transform Spectrometer (TANSO FTS) detects gas absorption spectra of the solar short wave infrared (SWIR) reflected on the Earth's surface as well as of the thermal infrared radiated from the ground and the atmosphere. TANSO-FTS is capable of detecting three narrow bands (0.76, 1.6, and 2.0 microm) and a wide band (5.5-14.3 microm) with 0.2 cm( 1) spectral resolution (interval). The TANSO Cloud and Aerosol Imager (TANSO-CAI) is an ultraviolet (UV), visible, near infrared, and SWIR radiometer designed to detect cloud and aerosol interference and to provide the data for their correction. GOSAT is placed in a sun-synchronous orbit 666 km at 13:00 local time, with an inclination angle of 98 degrees . A brief overview of the GOSAT project, scientific requirements, instrument designs, hardware performance, on orbit operation, and data processing is provided. PMID- 20011013 TI - Linear polarization difference imaging and its potential applications. AB - We demonstrate a novel linear polarization imaging technique and its potential application in dermatology. This technique records a series of images corresponding to different combinations of illumination and detection polarization and calculates intensity differences between orthogonal detection polarizations pixel by pixel. Fitting the polarization difference data to an analytical expression of the incident and detection polarization angles results in two new parameters, G and (phi3)/2. It is shown that G is strongly correlated to the order of alignment of the fibrous structure in the sample, and (phi3)/2 represents the angle of orientation of the fibers. Preliminary clinical testing implies that this method may be applied for medical diagnosis of skin diseases. PMID- 20011014 TI - Measurements of CO(2) concentration and temperature at high pressures using 1f normalized wavelength modulation spectroscopy with second harmonic detection near 2.7 microm. AB - Tunable diode lasers (TDL) near 2.7 mum are used to measure high-resolution direct absorption and wavelength modulation with second harmonic (WMS-2f) spectra at high pressures for two CO(2) transitions near 3633.08 and 3645.20 cm(-1), belonging to the nu(1)+ nu(3) vibrational band. Important factors influencing the design of a high-pressure TDL sensor and the variation of WMS-2f line shape with changes in pressure and laser parameters are discussed. Measurements of line strength and line broadening parameters are carried out for the 3645.20 cm(-1) transition in an atmospheric-pressure, high-temperature cell. A room-temperature high-pressure cell is then used to measure the pressure shift for both CO(2) transitions. Deviation of the direct absorption and wavelength modulation spectroscopy (WMS) spectra from the Lorentzian profile is studied in a high density (9.2 amagats) CO(2)-Ar mixture. The WMS spectra are shown to be negligibly affected by non-Lorentzian effects up to 10 atm and room temperature, in contrast with direct absorption. Measurements of CO(2) concentration and temperature are carried out in nonreactive shock-tube experiments (P approximately 8-12 atm, T~800-1200 K) to validate the accuracy and precision of wavelength-modulation-spectroscopy-based sensing. CO(2) time histories are then measured in heptane ignition experiments and compared with reaction kinetics mechanisms to demonstrate the use of this sensor in high-pressure combustion systems. PMID- 20011015 TI - Spectrally resolved cavity ring down measurement of high reflectivity mirrors using a supercontinuum laser source. AB - We investigate a cavity ring down setup that offers the possibility to measure the spectrally resolved reflectivities of high reflectivity mirrors. The setup consists of a resonator (ring down cavity) and an intensified CCD camera system combined with a spectrograph for spectral resolution. A commercial supercontinuum laser (350-1750 nm) is applied as a compact excitation source. It is based on a photonic crystal fiber that is pumped by a q-switched microchip laser (1.6 ns pulse duration, 25 kHz repetition rate). This combination allows simultaneously recording the transmittance of the cavity over a wide wavelength range determined by the excitation source and the spectral sensitivity of the detector. The photon lifetimes inside the cavity (ring down times) are measured with high spectral resolution by means of an intensified camera system. Subsequently shifting the "gate" of the image intensifier from short to long delay times after the excitation pulse allows calculation of the reflectivity spectrum of the mirrors. Comparison of these results with measurements using a conventional setup (laser diode 675 nm and photomultiplier tube) clearly shows the high potential of the method due to the multichannel excitation and the detection scheme. PMID- 20011016 TI - Flat-top steep-edge photodetector with cascaded grating structure. AB - A new type of photodetector with cascaded waveguide grating filters as a bottom reflection mirror is proposed. A greatly improved spectral response is shown to follow by the integration of a waveguide grating into classical thin-film homogeneous layers. Calculation results for a single grating, a cascaded-double grating, and a cascaded-triple grating structure are demonstrated. An increasing rectangular spectral response is obtained by cascading two or three grating filters. Compared with a traditional photodetector with distributed Bragg reflectors, this new type of photodetector with the same materials requires significantly fewer layers while maintaining narrow flattop response, high peak efficiency, and low sideband reflectance. PMID- 20011017 TI - Fabrication and characterization of infrared hollow fiber with multi- SiO(2) and AgI inner-coating layers. AB - We report the transmission characteristics of infrared hollow fiber with multi- AgI and SiO(2) inner-coating layers in the mid-infrared region. A three dielectric-layer hollow glass fiber with a SiO(2)-AgI-SiO(2)-Ag structure was fabricated and low-loss property was obtained in the mid-infrared region. The SiO(2) films were coated by use of the liquid-phase coating method and a semi inorganic polymer was used as the coating material. For deposition of the AgI film between the two SiO(2) films, a silver film was first plated by use of the silver mirror reaction method. Then the iodination process was conducted to turn the silver layer into silver iodide. A calculation method was also developed to estimate the film thickness of dielectric layers in each fabrication step according to the position of loss peaks in the measured loss spectra. Good agreement between calculated and measured loss spectra was demonstrated by taking into consideration material dispersion and surface roughness of inner-coating films. PMID- 20011018 TI - Linear regression models and neural networks for the fast emulation of a molecular absorption code. AB - The background scene generator MATISSE, whose main functionality is to generate natural background radiance images, makes use of the so-called Correlated K (CK) model. It necessitates either loading or computing thousands of CK coefficients for each atmospheric profile. When the CK coefficients cannot be loaded, the computation time becomes prohibitive. The idea developed in this paper is to substitute fast approximate models for the exact CK generator; using the latter, a representative set of numerical examples is built and used to train linear or nonlinear regression models. The resulting models enable an accurate CK coefficient computation for all the profiles of an image in a reasonable time. PMID- 20011019 TI - Nonlinear absorption in single LaF(3) and MgF(2) layers at 193 nm measured by surface sensitive laser induced deflection technique. AB - We report nonlinear absorption data of LaF(3) and MgF(2) single layers at 193 nm. A highly surface sensitive measurement strategy of the laser induced deflection technique is introduced and applied to measure the absorption of highly transparent thin films independently of the substrate absorption. Linear absorptions k=(alphaxlambda)/4pi of 2x10(-4) and 8.5x10(-4) (LaF(3)) and 1.8x10( 4) and 6.9x10(-4) (MgF(2)) are found. Measured two photon absorption (TPA) coefficients are beta=1x10(-4) cm/W (LaF(3)), 1.8x10(-5), and 5.8x10(-5) cm/W (MgF(2)). The TPA coefficients are several orders of magnitude higher than typical values for fluoride single crystals, which is likely to result from sequential two step absorption processes. PMID- 20011020 TI - Design of error-compensating algorithms for sinusoidal phase shifting interferometry. AB - An improved approach to interferometry using sinusoidal phase shifting balances several harmonic components in the interference signal against each other. The resulting computationally efficient phase-estimation algorithms have low sensitivity to errors such as spurious intensity noise, vibration, and errors in the phase shift pattern. Specific example algorithms employing 8 and 12 camera frames illustrate design principles that are extendable to algorithms of any length for applications that would benefit from a simplified, sinusoidal phase shift. PMID- 20011021 TI - Zirconia-coated carbonyl-iron-particle-based magnetorheological fluid for polishing optical glasses and ceramics. AB - We report on magnetorheological finishing (MRF) spotting experiments performed on glasses and ceramics using a zirconia-coated carbonyl-iron (CI)-particle-based magnetorheological (MR) fluid. The zirconia-coated magnetic CI particles were prepared via sol-gel synthesis in kilogram quantities. The coating layer was approximately 50-100 nm thick, faceted in surface structure, and well adhered. Coated particles showed long-term stability against aqueous corrosion. "Free" nanocrystalline zirconia polishing abrasives were cogenerated in the coating process, resulting in an abrasive-charged powder for MRF. A viable MR fluid was prepared simply by adding water. Spot polishing tests were performed on a variety of optical glasses and ceramics over a period of nearly three weeks with no signs of MR fluid degradation or corrosion. Stable material removal rates and smooth surfaces inside spots were obtained. PMID- 20011022 TI - Angular shape of the oceanic particulate volume scattering function in the backward direction. AB - Analysis of several million particulate volume scattering functions (VSFs) from different field sites around the world's oceans and coastlines revealed that the shape of the VSF in the backward direction was remarkably consistent (5% or less variability at angles between 90 degrees and 170 degrees ). In agreement with theoretical models and past field measurements, the variability of the VSF shape (the VSF normalized to the backscattering coefficient) was found to be lowest between 110 degrees and 120 degrees . This study concludes that under most oceanic conditions, estimates of the particulate backscattering coefficient, using single angle scattering measurements near 110 degrees to 120 degrees and suitable conversion factors, are justified and should have a maximum uncertainty of less than a few percent once instrument noise is accounted for. PMID- 20011023 TI - Optimizing photophoresis and asymmetric force fields for grading of Brownian particles. AB - We discuss a scheme that incorporates restricted spatial input location, orthogonal sort, and movement direction features, with particle sorting achieved by using an asymmetric potential cycled on and off, while movement is accomplished by photophoresis. Careful investigation has uncovered the odds of sorting between certain pairs of particle sizes to be solely dependent on radii in each phase of the process. This means that the most effective overall sorting can be achieved by maximizing the number of phases. This optimized approach is demonstrated using numerical simulation to permit grading of a range of nanometer scale particle sizes. PMID- 20011024 TI - Tunable ring laser using a tapered single mode fiber tip. AB - A tunable ring laser using a tapered single mode fiber tip as a bandpass filter has been proposed and demonstrated for the first time to our knowledge. This is a simple and cost-effective tunable source. It is found that the tuning range and bandwidth of the laser are related to the relaxation time of the optical amplifier, the current of the amplifier, and the steepness of the tip shape. The calculations and experimental results show that the laser has a tuning range of 9 nm in the L-band and the spectral linewidth can be varied from 0.06 nm to 0.17 nm. PMID- 20011025 TI - Efficient approach to designing a Schmidt-Cassegrain objective for a remote sensing satellite. AB - This paper presents an efficient approach to designing a Schmidt-Cassegrain objective for a remote sensing satellite. The objective is required to have multispectral operational bands, with three spectral channels distributed along the range (0.5 to 0.9 mum), as well as a panchromatic channel; 4 degrees field of view; distortion smaller than 0.3%; and a modulation transfer function, at 50 lines/mm spatial frequency, better than 0.5 and 0.35 at the center and edge of the field of view. The proposed design approach is based on Slyusarev's theory of aberrations and optical design. An image quality index is formulated as a function of optical system component powers and axial distances. For each combination of parameters, there exists a possible solution that can be realized into a thin lens system by solving Seidel sum equations. The final design is then reached by a simple and quick optimization step. The best three designs are compared in terms of initial values of optical system parameters and final design specifications. The best system image quality is thoroughly analyzed. All three presented designs meet and exceed the required design specifications. PMID- 20011026 TI - Concise assembly of linear alpha(1-->6)-linked octamannan fluorescent probe. AB - Synthesis of a fluorescently labelled (dansylated) linear alpha(1-->6)-linked octamannan, using glycosyl fluoride donors and thioglycosyl acceptors is described. A selective and convergent two-stage activation progression was executed to construct di-, tetra and octa-mannosyl thioglycosides in three glycosylation steps with excellent yield. Further a 5-N,N Dimethylaminonaphthalene-1-sulfonamidoethyl (dansyl) group was coupled to 1 azidoethyl octamannosyl thioglycoside. Global deprotection of the coupled product afforded the desired dansylated homo-linear alpha(1-->6)-linked octamannan. PMID- 20011027 TI - Highly efficient palladium-catalyzed hydrostannation of ethyl ethynyl ether. AB - The palladium-catalyzed hydrostannation of acetylenes is widely exploited in organic synthesis as a means of forming vinyl stannanes for use in palladium catalyzed cross-coupling reactions. Application of this methodology to ethyl ethynyl ether results in an enol ether that is challenging to isolate from the crude reaction mixture because of incompatibility with typical silica gel chromatography. Reported here is a highly efficient procedure for the palladium catalyzed hydrostannation of ethyl ethynyl ether using 0.1% palladium(0) catalyst and 1.0 equiv of tributyltin hydride. The product obtained is a mixture of regioisomers that can be carried forward with exclusive reaction of the beta isomer. This method is highly reproducible; relative to previously reported procedures, it is more economical and involves a more facile purification procedure. PMID- 20011028 TI - (3R, 9R, 10R)-Panaxytriol: A molecular-based nutraceutical with possible application to cancer prevention and treatment. AB - Panaxytriol is a nutraceutical-based active constituent of Korean red ginseng and is reported to exhibit potent anti-tumor properties. Its activity may be in part due to its induction of phase 2 chemoprotective enzymes. Its unique properties may have important implications in cancer therapeutics. PMID- 20011029 TI - Carbon-rich supramolecular metallacycles and metallacages. AB - Coordination-driven self-assembly via the directional-bonding approach utilizes rigid transition metal acceptors and electron-rich donors to allow for complex, nanoscale 2D polygons and 3D polyhedra to be prepared under mild conditions and in high yields. To ensure proper rigidity and directionality, many acceptor and donor precursors contain largely carbon-rich aromatic and/or acetylenic moieties. This article introduces self-assembly as an alternative means of synthesizing carbon-rich materials and discusses the development, design, synthesis, and applications of carbon-rich supramolecular metallacycles and metallacages as well as the self-assembly of new diastereomeric carbon-rich supramolecular triangles. PMID- 20011030 TI - Design and synthesis of water-soluble bioconjugatable trans-AB-porphyrins. AB - Three free base porphyrins have been prepared that bear a polar and facially encumbering 2,4,6-tris(carboxymethoxy)phenyl motif at one meso (5-) position. The only other substituent (15-position) comprises phenyl, formyl, or p-aminophenyl. The porphyrins exhibit solubility in water (or aqueous buffer solutions) at pH >/=7 and concentrations >1 mM at room temperature. The concise syntheses, water solubility, and bioconjugatable handle make these porphyrin constructs suitable for biological applications. PMID- 20011031 TI - Congenital desmoid tumor of the cheek: a clinicopathological case report. AB - OBJECTIVE: Desmoid tumors are rare benign neoplasms of fibroblastic origin, frequently presenting in the abdomen of adult patients. Little is reported about clinical features and outcome of desmoid tumors appearing in infants and children, especially when they are located in the face. METHODS: We report a girl with a desmoid tumor of her cheek, which was present at birth. RESULTS: The tumor was treated by surgical excision, with no additional treatment, other than follow up. No recurrence of the tumor was witnessed 15 months following surgical excision. SUMMARY: Radical surgical treatment with tumor excision, sometimes accompanied by radiotherapy, is the current standard therapy for infantile desmoid tumors. Pharmacological treatment may be added in selected cases. These modalities may be challenging when desmoid tumors appear in the face because of risks of facial distortion and associated growth problems. PMID- 20011032 TI - White shark offshore habitat: a behavioral and environmental characterization of the eastern Pacific shared offshore foraging area. AB - BACKGROUND: Although much is known about the behavior of white sharks in coastal regions, very little is known about their vertical movements offshore in the eastern Pacific where they spend up to five months. We provide the first detailed description of the offshore habitat use of white sharks in the eastern North Pacific. METHODOLOGY/PRINCIPAL FINDINGS: This study uses 2-min data from four recovered pop-up satellite archival tags deployed at Guadalupe Island (2002 and 2005). Deployments ranged from 5.4 to 8.2 months. Two predominant vertical patterns were described. The first was a bimodal vertical pattern with time spent at the surface and at depth, which was observed while traveling. The second was a repetitive oscillatory diving mode displayed by sharks in the Shared Offshore Foraging Area (SOFA). For all four datasets the average maximum daily dive depths ranged from 442.5 to 492.8 m and were typically associated with dissolved oxygen concentrations of above 1.7 ml L(-1). Although infrequent, occasional dives to near 1000 m with a minimum temperature of 3.9 degrees C and a minimum O(2) level of 0.3 ml L(-1) were observed. CONCLUSIONS/SIGNIFICANCE: Recovered pop-up satellite tags from Guadalupe Island white sharks advance our understanding of the vertical habitat use of white sharks while offshore. The bimodal vertical pattern during traveling is most likely related to geolocation. The oscillatory dive pattern is likely associated with foraging. While feeding is not documented, foraging is likely occurring in association with the deep scattering layer. Diving depths were not limited by temperature but were constrained by O(2) levels below approximately 1.5 ml L(-1). While oxygen may limit the extent of sharks' vertical movements, it will also impact prey distribution. Consequently, the shallow oxygen minimum zone in the SOFA may act to concentrate prey, thus enhancing foraging opportunities in these oligotrophic waters. PMID- 20011033 TI - Exploring cortical attentional system by using fMRI during a Continuous Perfomance Test. AB - Functional magnetic resonance imaging (fMRI) was performed in eight healthy subjects to identify the localization, magnitude, and volume extent of activation in brain regions that are involved in blood oxygen level-dependent (BOLD) response during the performance of Conners' Continuous Performance Test (CPT). An extensive brain network was activated during the task including frontal, temporal, and occipital cortical areas and left cerebellum. The more activated cluster in terms of volume extent and magnitude was located in the right anterior cingulate cortex (ACC). Analyzing the dynamic trend of the activation in the identified areas during the entire duration of the sustained attention test, we found a progressive decreasing of BOLD response probably due to a habituation effect without any deterioration of the performances. The observed brain network is consistent with existing models of visual object processing and attentional control and may serve as a basis for fMRI studies in clinical populations with neuropsychological deficits in Conners' CPT performance. PMID- 20011034 TI - A wireless brain-machine interface for real-time speech synthesis. AB - BACKGROUND: Brain-machine interfaces (BMIs) involving electrodes implanted into the human cerebral cortex have recently been developed in an attempt to restore function to profoundly paralyzed individuals. Current BMIs for restoring communication can provide important capabilities via a typing process, but unfortunately they are only capable of slow communication rates. In the current study we use a novel approach to speech restoration in which we decode continuous auditory parameters for a real-time speech synthesizer from neuronal activity in motor cortex during attempted speech. METHODOLOGY/PRINCIPAL FINDINGS: Neural signals recorded by a Neurotrophic Electrode implanted in a speech-related region of the left precentral gyrus of a human volunteer suffering from locked-in syndrome, characterized by near-total paralysis with spared cognition, were transmitted wirelessly across the scalp and used to drive a speech synthesizer. A Kalman filter-based decoder translated the neural signals generated during attempted speech into continuous parameters for controlling a synthesizer that provided immediate (within 50 ms) auditory feedback of the decoded sound. Accuracy of the volunteer's vowel productions with the synthesizer improved quickly with practice, with a 25% improvement in average hit rate (from 45% to 70%) and 46% decrease in average endpoint error from the first to the last block of a three-vowel task. CONCLUSIONS/SIGNIFICANCE: Our results support the feasibility of neural prostheses that may have the potential to provide near conversational synthetic speech output for individuals with severely impaired speech motor control. They also provide an initial glimpse into the functional properties of neurons in speech motor cortical areas. PMID- 20011035 TI - Long-term stability of visual pattern selective responses of monkey temporal lobe neurons. AB - Many neurons in primate inferotemporal (IT) cortex respond selectively to complex, often meaningful, stimuli such as faces and objects. An important unanswered question is whether such response selectivity, which is thought to arise from experience-dependent plasticity, is maintained from day to day, or whether the roles of individual cells are continually reassigned based on the diet of natural vision. We addressed this question using microwire electrodes that were chronically implanted in the temporal lobe of two monkeys, often allowing us to monitor activity of individual neurons across days. We found that neurons maintained their selectivity in both response magnitude and patterns of spike timing across a large set of visual images throughout periods of stable signal isolation from the same cell that sometimes exceeded two weeks. These results indicate that stimulus-selectivity of responses in IT is stable across days and weeks of visual experience. PMID- 20011037 TI - Cluster designs to assess the prevalence of acute malnutrition by lot quality assurance sampling: a validation study by computer simulation. AB - Traditional lot quality assurance sampling (LQAS) methods require simple random sampling to guarantee valid results. However, cluster sampling has been proposed to reduce the number of random starting points. This study uses simulations to examine the classification error of two such designs, a 67x3 (67 clusters of three observations) and a 33x6 (33 clusters of six observations) sampling scheme to assess the prevalence of global acute malnutrition (GAM). Further, we explore the use of a 67x3 sequential sampling scheme for LQAS classification of GAM prevalence. Results indicate that, for independent clusters with moderate intracluster correlation for the GAM outcome, the three sampling designs maintain approximate validity for LQAS analysis. Sequential sampling can substantially reduce the average sample size that is required for data collection. The presence of intercluster correlation can impact dramatically the classification error that is associated with LQAS analysis. PMID- 20011036 TI - Lymphatic vascularisation and involvement of Lyve-1+ macrophages in the human onchocerca nodule. AB - Onchocerciasis, caused by the filarial nematode Onchocerca volvulus, is a parasitic disease leading to debilitating skin disease and blindness, with major economic and social consequences. The pathology of onchocerciasis is principally considered to be a consequence of long-standing host inflammatory responses. In onchocerciasis a subcutaneous nodule is formed around the female worms, the core of which is a dense infiltrate of inflammatory cells in which microfilariae are released. It has been established that the formation of nodules is associated with angiogenesis. In this study, we show using specific markers of endothelium (CD31) and lymphatic endothelial cells (Lyve-1, Podoplanin) that not only angiogenesis but also lymphangiogenesis occurs within the nodule. 7% of the microfilariae could be found within the lymphatics, but none within blood vessels in these nodules, suggesting a possible route of migration for the larvae. The neovascularisation was associated with a particular pattern of angio/lymphangiogenic factors in nodules of onchocerciasis patients, characterized by the expression of CXCL12, CXCR4, VEGF-C, Angiopoietin-1 and Angiopoietin-2. Interestingly, a proportion of macrophages were found to be positive for Lyve-1 and some were integrated into the endothelium of the lymphatic vessels, revealing their plasticity in the nodular micro-environment. These results indicate that lymphatic as well as blood vascularization is induced around O. volvulus worms, either by the parasite itself, e.g. by the release of angiogenic and lymphangiogenic factors, or by consecutive host immune responses. PMID- 20011038 TI - Estimating and testing haplotype-trait associations in non-diploid populations. AB - Malaria is an infectious disease that is caused by a group of parasites of the genus Plasmodium. Characterizing the association between polymorphisms in the parasite genome and measured traits in an infected human host may provide insight into disease aetiology and ultimately inform new strategies for improved treatment and prevention. This, however, presents an analytic challenge since individuals are often multiply infected with a variable and unknown number of genetically diverse parasitic strains. In addition, data on the alignment of nucleotides on a single chromosome, which is commonly referred to as haplotypic phase, is not generally observed. An expectation-maximization algorithm for estimating and testing associations between haplotypes and quantitative traits has been described for diploid (human) populations. We extend this method to account for both the uncertainty in haplotypic phase and the variable and unknown number of infections in the malaria setting. Further extensions are described for the human immunodeficiency virus quasi-species setting. A simulation study is presented to characterize performance of the method. Application of this approach to data arising from a cross-sectional study of n=126 multiply infected children in Uganda reveals some interesting associations requiring further investigation. PMID- 20011039 TI - Bone T-scores and functional status: a cross-sectional study on German elderly. AB - BACKGROUND: We explore the association between bone T-scores, used in osteoporosis diagnosis, and functional status since we hypothesized that bone health can impact elderly functional status and indirectly independence. METHODS: In a cross-sectional study (2005-2006) on community dwelling elderly (> = 75 years) from Herne, Germany we measured bone T-scores with Dual-energy X-ray Absorptiometry, and functional status indexed by five geriatric tests: activities of daily living, instrumental activities of daily living, test of dementia, geriatric depression score and the timed-up-and-go test, and two pooled indexes: raw and standardized. Generalized linear regression was used to determine the relationship between T-scores and functional status. RESULTS: From 3243 addresses, only 632 (19%) completed a clinical visit, of which only 440 (male:female, 243:197) could be included in analysis. T-scores (-0.99, 95% confidence interval [CI], -1.1-0.9) predicted activities of daily living (95.3 CI, 94.5-96.2), instrumental activities of daily living (7.3 CI, 94.5-96.2), and timed-up-and-go test (10.7 CI, 10.0-11.3) (P < = 0.05). Pooled data showed that a unit improvement in T-score improved standardized pooled functional status (15 CI, 14.7-15.3) by 0.41 and the raw (99.4 CI, 97.8-101.0) by 2.27 units. These results were limited due to pooling of different scoring directions, selection bias, and a need to follow-up with evidence testing. CONCLUSIONS: T-scores associated with lower functional status in community-dwelling elderly. Regular screening of osteoporosis as a preventive strategy might help maintain life quality with aging. PMID- 20011040 TI - Glimepiride reduces the expression of PrPc, prevents PrPSc formation and protects against prion mediated neurotoxicity in cell lines. AB - BACKGROUND: A hallmark of the prion diseases is the conversion of the host encoded cellular prion protein (PrP(C)) into a disease related, alternatively folded isoform (PrP(Sc)). The accumulation of PrP(Sc) within the brain is associated with synapse loss and ultimately neuronal death. Novel therapeutics are desperately required to treat neurodegenerative diseases including the prion diseases. PRINCIPAL FINDINGS: Treatment with glimepiride, a sulphonylurea approved for the treatment of diabetes mellitus, induced the release of PrP(C) from the surface of prion-infected neuronal cells. The cell surface is a site where PrP(C) molecules may be converted to PrP(Sc) and glimepiride treatment reduced PrP(Sc) formation in three prion infected neuronal cell lines (ScN2a, SMB and ScGT1 cells). Glimepiride also protected cortical and hippocampal neurones against the toxic effects of the prion-derived peptide PrP82-146. Glimepiride treatment significantly reduce both the amount of PrP82-146 that bound to neurones and PrP82-146 induced activation of cytoplasmic phospholipase A(2) (cPLA(2)) and the production of prostaglandin E(2) that is associated with neuronal injury in prion diseases. Our results are consistent with reports that glimepiride activates an endogenous glycosylphosphatidylinositol (GPI) phospholipase C which reduced PrP(C) expression at the surface of neuronal cells. The effects of glimepiride were reproduced by treatment of cells with phosphatidylinositol-phospholipase C (PI-PLC) and were reversed by co-incubation with p-chloromercuriphenylsulphonate, an inhibitor of endogenous GPI-PLC. CONCLUSIONS: Collectively, these results indicate that glimepiride may be a novel treatment to reduce PrP(Sc) formation and neuronal damage in prion diseases. PMID- 20011041 TI - Extramitochondrial Ca2+ in the nanomolar range regulates glutamate-dependent oxidative phosphorylation on demand. AB - We present unexpected and novel results revealing that glutamate-dependent oxidative phosphorylation (OXPHOS) of brain mitochondria is exclusively and efficiently activated by extramitochondrial Ca(2+) in physiological concentration ranges (S(0.5) = 360 nM Ca(2+)). This regulation was not affected by RR, an inhibitor of the mitochondrial Ca(2+) uniporter. Active respiration is regulated by glutamate supply to mitochondria via aralar, a mitochondrial glutamate/aspartate carrier with regulatory Ca(2+)-binding sites in the mitochondrial intermembrane space providing full access to cytosolic Ca(2+). At micromolar concentrations, Ca(2+) can also enter the intramitochondrial matrix and activate specific dehydrogenases. However, the latter mechanism is less efficient than extramitochondrial Ca(2+) regulation of respiration/OXPHOS via aralar. These results imply a new mode of glutamate-dependent OXPHOS regulation as a demand-driven regulation of mitochondrial function. This regulation involves the mitochondrial glutamate/aspartate carrier aralar which controls mitochondrial substrate supply according to the level of extramitochondrial Ca(2+). PMID- 20011042 TI - A cellular automata-based mathematical model for thymocyte development. AB - Intrathymic T cell development is an important process necessary for the normal formation of cell-mediated immune responses. Importantly, such a process depends on interactions of developing thymocytes with cellular and extracellular elements of the thymic microenvironment. Additionally, it includes a series of oriented and tunely regulated migration events, ultimately allowing mature cells to cross endothelial barriers and leave the organ. Herein we built a cellular automata based mathematical model for thymocyte migration and development. The rules comprised in this model take into account the main stages of thymocyte development, two-dimensional sections of the normal thymic microenvironmental network, as well as the chemokines involved in intrathymic cell migration. Parameters of our computer simulations with further adjusted to results derived from previous experimental data using sub-lethally irradiated mice, in which thymus recovery can be evaluated. The model fitted with the increasing numbers of each CD4/CD8-defined thymocyte subset. It was further validated since it fitted with the times of permanence experimentally ascertained in each CD4/CD8-defined differentiation stage. Importantly, correlations using the whole mean volume of young normal adult mice revealed that the numbers of cells generated in silico with the mathematical model fall within the range of total thymocyte numbers seen in these animals. Furthermore, simulations made with a human thymic epithelial network using the same mathematical model generated similar profiles for temporal evolution of thymocyte developmental stages. Lastly, we provided in silico evidence that the thymus architecture is important in the thymocyte development, since changes in the epithelial network result in different theoretical profiles for T cell development/migration. This model likely can be used to predict thymocyte evolution following therapeutic strategies designed for recovery of the thymus in diseases coursing with thymus involution, such as some primary immunodeficiencies, acute infections, and malnutrition. PMID- 20011043 TI - Cigarette smoking blocks the protective expression of Nrf2/ARE pathway in peripheral mononuclear cells of young heavy smokers favouring inflammation. AB - Cigarette smoking is an important risk factor for atherosclerosis, a chronic inflammatory disease. However the underlying factors of this effect are unclear. It has been hypothesized that water-soluble components of cigarette smoke can directly promote oxidative stress in vasculature and blood cells. Aim of this study was to study the relationship between oxidative stress and inflammation in a group of young smokers. To do this we evaluated: 1) the oxidation products of phospholipids (oxPAPC) in peripheral blood mononuclear cells (PBMC); 2) their role in causing PBMC reactive oxygen species (ROS) generation and changes in GSH; 3) the expression of the transcription factor NF-E2-related factor 2 (Nrf2) and of related antioxidant genes (ARE); 4) the activation of NF-kB and C-reactive protein (CRP) values. We studied 90 healthy volunteers: 32 non-smokers, 32 moderate smokers (5-10 cigarettes/day) and 26 heavy smokers (25-40 cigarettes/day). OxPAPC and p47phox expression, that reasonably reflects NADPH oxidase activity, were higher in moderate smokers and heavy smokers than in non smokers (p<0.01), the highest values being in heavy smokers (p<0.01). In in vitro studies oxPAPC increased ROS generation via NADPH oxidase activation. GSH in PBMC and plasma was lower in moderate smokers and heavy smokers than in non-smokers (p<0.01), the lowest values being in heavy smokers (p<0.01). Nrf2 expression in PBMC was higher in moderate smokers than in non-smokers (p<0.01), but not in heavy smokers, who had the highest levels of NF-kB and CRP (p<0.01). In in vitro studies oxPAPC dose-dependently increased NF-kB activation, whereas at the highest concentrations Nrf2 expression was repressed. The small interference (si) RNA-mediated knockdown of NF-kappaB/p65 increased about three times the expression of Nrf2 stimulated with oxPAPC. Cigarette smoke promotes oxPAPC formation and oxidative stress in PBMC. This may cause the activation of NF-kB that in turn may participate in the negative regulation of Nrf2/ARE pathway favouring inflammation. PMID- 20011044 TI - Using association mapping in teosinte to investigate the function of maize selection-candidate genes. AB - BACKGROUND: Large-scale screens of the maize genome identified 48 genes that show the putative signature of artificial selection during maize domestication or improvement. These selection-candidate genes may act as quantitative trait loci (QTL) that control the phenotypic differences between maize and its progenitor, teosinte. The selection-candidate genes appear to be located closer in the genome to domestication QTL than expected by chance. METHODS AND FINDINGS: As a step toward defining the traits controlled by these genes, we performed phenotype genotype association mapping in teosinte for 32 of the 48 plus three other selection-candidate genes. Our analyses assayed 32 phenotypic traits, many of which were altered during maize domestication or improvement. We observed several significant associations between SNPs in the selection-candidate genes and trait variation in teosinte. These included two associations that surpassed the Bonferroni correction and five instances where a gene significantly associated with the same trait in both of our association mapping panels. Despite these significant associations, when compared as a group the selection-candidate genes performed no better than randomly chosen genes. CONCLUSIONS: Our results suggest association analyses can be helpful for identifying traits under the control of selection-candidate genes. Indeed, we present evidence for new functions for several selection-candidate genes. However, with the current set of selection candidate genes and our association mapping strategy, we found very few significant associations overall and no more than we would have found with randomly chosen genes. We discuss possible reasons that a large number of significant genotype-phenotype associations were not discovered. PMID- 20011045 TI - Toll-like receptor mRNA expression is selectively increased in the colonic mucosa of two animal models relevant to irritable bowel syndrome. AB - BACKGROUND: Irritable bowel syndrome (IBS) is largely viewed as a stress-related disorder caused by aberrant brain-gut-immune communication and altered gastrointestinal (GI) homeostasis. Accumulating evidence demonstrates that stress modulates innate immune responses; however, very little is known on the immunological effects of stress on the GI tract. Toll-like receptors (TLRs) are critical pattern recognition molecules of the innate immune system. Activation of TLRs by bacterial and viral molecules leads to activation of NF-kB and an increase in inflammatory cytokine expression. It was our hypothesis that innate immune receptor expression may be changed in the gastrointestinal tract of animals with stress-induced IBS-like symptoms. METHODOLOGY/PRINCIPAL FINDINGS: In this study, our objective was to evaluate the TLR expression profile in the colonic mucosa of two rat strains that display colonic visceral hypersensitivity; the stress-sensitive Wistar-Kyoto (WKY) rat and the maternally separated (MS) rat. Quantitative PCR of TLR2-10 mRNA in both the proximal and distal colonic mucosae was carried out in adulthood. Significant increases are seen in the mRNA levels of TLR3, 4 & 5 in both the distal and proximal colonic mucosa of MS rats compared with controls. No significant differences were noted for TLR 2, 7, 9 & 10 while TLR 6 could not be detected in any samples in both rat strains. The WKY strain have increased levels of mRNA expression of TLR3, 4, 5, 7, 8, 9 & 10 in both the distal and proximal colonic mucosa compared to the control Sprague Dawley strain. No significant differences in expression were found for TLR2 while as before TLR6 could not be detected in all samples in both strains. CONCLUSIONS: These data suggest that both early life stress (MS) and a genetic predisposition (WKY) to stress affect the expression of key sentinels of the innate immune system which may have direct relevance for the molecular pathophysiology of IBS. PMID- 20011046 TI - Bayesian receiver operating characteristic estimation of multiple tests for diagnosis of bovine tuberculosis in Chadian cattle. AB - BACKGROUND: Bovine tuberculosis (BTB) today primarily affects developing countries. In Africa, the disease is present essentially on the whole continent; however, little accurate information on its distribution and prevalence is available. Also, attempts to evaluate diagnostic tests for BTB in naturally infected cattle are scarce and mostly complicated by the absence of knowledge of the true disease status of the tested animals. However, diagnostic test evaluation in a given setting is a prerequisite for the implementation of local surveillance schemes and control measures. METHODOLOGY/PRINCIPAL FINDINGS: We subjected a slaughterhouse population of 954 Chadian cattle to single intra dermal comparative cervical tuberculin (SICCT) testing and two recently developed fluorescence polarization assays (FPA). Using a Bayesian modeling approach we computed the receiver operating characteristic (ROC) curve of each diagnostic test, the true disease prevalence in the sampled population and the disease status of all sampled animals in the absence of knowledge of the true disease status of the sampled animals. In our Chadian setting, SICCT performed better if the cut-off for positive test interpretation was lowered from >4 mm (OIE standard cut-off) to >2 mm. Using this cut-off, SICCT showed a sensitivity and specificity of 66% and 89%, respectively. Both FPA tests showed sensitivities below 50% but specificities above 90%. The true disease prevalence was estimated at 8%. Altogether, 11% of the sampled animals showed gross visible tuberculous lesions. However, modeling of the BTB disease status of the sampled animals indicated that 72% of the suspected tuberculosis lesions detected during standard meat inspections were due to other pathogens than Mycobacterium bovis. CONCLUSIONS/SIGNIFICANCE: Our results have important implications for BTB diagnosis in a high incidence sub-Saharan African setting and demonstrate the practicability of our Bayesian approach for diagnostic test evaluation. PMID- 20011047 TI - Sub-optimal allocation of time in sequential movements. AB - The allocation of limited resources such as time or energy is a core problem that organisms face when planning complex actions. Most previous research concerning planning of movement has focused on the planning of single, isolated movements. Here we investigated the allocation of time in a pointing task where human subjects attempted to touch two targets in a specified order to earn monetary rewards. Subjects were required to complete both movements within a limited time but could freely allocate the available time between the movements. The time constraint presents an allocation problem to the subjects: the more time spent on one movement, the less time is available for the other. In different conditions we assigned different rewards to the two tokens. How the subject allocated time between movements affected their expected gain on each trial. We also varied the angle between the first and second movements and the length of the second movement. Based on our results, we developed and tested a model of speed-accuracy tradeoff for sequential movements. Using this model we could predict the time allocation that would maximize the expected gain of each subject in each experimental condition. We compared human performance with predicted optimal performance. We found that all subjects allocated time sub-optimally, spending more time than they should on the first movement even when the reward of the second target was five times larger than the first. We conclude that the movement planning system fails to maximize expected reward in planning sequences of as few as two movements and discuss possible interpretations drawn from economic theory. PMID- 20011048 TI - Unconsciously perceived fear in peripheral vision alerts the limbic system: a MEG study. AB - BACKGROUND: In ecological situations, threatening stimuli often come out from the peripheral vision. Such aggressive messages must trigger rapid attention to the periphery to allow a fast and adapted motor reaction. Several clues converge to hypothesize that peripheral danger presentation can trigger off a fast arousal network potentially independent of the consciousness spot. METHODOLOGY/PRINCIPAL FINDINGS: In the present MEG study, spatio-temporal dynamics of the neural processing of danger related stimuli were explored as a function of the stimuli position in the visual field. Fearful and neutral faces were briefly presented in the central or peripheral visual field, and were followed by target faces stimuli. An event-related beamformer source analysis model was applied in three time windows following the first face presentations: 80 to 130 ms, 140 to 190 ms, and 210 to 260 ms. The frontal lobe and the right internal temporal lobe part, including the amygdala, reacted as soon as 80 ms of latency to fear occurring in the peripheral vision. For central presentation, fearful faces evoked the classical neuronal activity along the occipito-temporal visual pathway between 140 and 190 ms. CONCLUSIONS: Thus, the high spatio-temporal resolution of MEG allowed disclosing a fast response of a network involving medial temporal and frontal structures in the processing of fear related stimuli occurring unconsciously in the peripheral visual field. Whereas centrally presented stimuli are precisely processed by the ventral occipito-temporal cortex, the related-to danger stimuli appearing in the peripheral visual field are more efficient to produce a fast automatic alert response possibly conveyed by subcortical structures. PMID- 20011049 TI - Androgen receptor mutations associated with androgen insensitivity syndrome: a high content analysis approach leading to personalized medicine. AB - Androgen insensitivity syndrome (AIS) is a rare disease associated with inactivating mutations of AR that disrupt male sexual differentiation, and cause a spectrum of phenotypic abnormalities having as a common denominator loss of reproductive viability. No established treatment exists for these conditions, however there are sporadic reports of patients (or recapitulated mutations in cell lines) that respond to administration of supraphysiologic doses (or pulses) of testosterone or synthetic ligands. Here, we utilize a novel high content analysis (HCA) approach to study AR function at the single cell level in genital skin fibroblasts (GSF). We discuss in detail findings in GSF from three historical patients with AIS, which include identification of novel mechanisms of AR malfunction, and the potential ability to utilize HCA for personalized treatment of patients affected by this condition. PMID- 20011050 TI - Immunization of mice with a recombinant adenovirus vaccine inhibits the early growth of Mycobacterium tuberculosis after infection. AB - BACKGROUND: In pulmonary Mycobacterium tuberculosis (Mtb) infection, immune responses are delayed compared to other respiratory infections, so that antigen specific cells are not detected in the lungs earlier than day 14. Even after parenteral immunization with Bacille Calmette Guerin (BCG) or a subunit vaccine, the immune response after Mtb challenge is only slightly accelerated and the kinetics of pulmonary Mtb growth do not differ between naive and immunized animals up to day 14. METHODS AND FINDINGS: Mice were immunized intranasally with a recombinant adenovirus expressing mycobacterial antigen 85A (Ad85A), challenged by aerosol with Mtb and the kinetics of Mtb growth in the lungs measured. Intranasal immunization with Ad85A inhibits Mtb growth in the early phase of infection, up to day 8. Protection is sustained for at least 7 months and correlates with the presence of antigen-specific activated effector CD8 T cells in the lungs. Antigen 85A-specific T cells respond to antigen presenting cells from the lungs of mice immunized with Ad85A 23 weeks previously, demonstrating the persistence of antigen in the lungs. CONCLUSIONS/SIGNIFICANCE: Intranasal immunization with Ad85A can inhibit early growth of Mtb because it establishes a lung antigen depot and maintains an activated lung-resident lymphocyte population. We propose that an optimal immunization strategy for tuberculosis should aim to induce both lung and systemic immunity, targeting the early and late phases of Mtb growth. PMID- 20011051 TI - The immunogenic SigA enterotoxin of Shigella flexneri 2a binds to HEp-2 cells and induces fodrin redistribution in intoxicated epithelial cells. AB - BACKGROUND: We have previously shown that the enterotoxin SigA which resides on the she pathogenicity island (PAI) of S. flexneri 2a is an autonomously secreted serine protease capable of degrading casein. We have also demonstrated that SigA is cytopathic for HEp-2 cells and plays a role in the intestinal fluid accumulation associated with S. flexneri infections. METHODS/PRINCIPAL FINDINGS: In this work we show that SigA binds specifically to HEp-2 cells and degrades recombinant human alphaII spectrin (alpha-fodrin) in vitro, suggesting that the cytotoxic and enterotoxic effects mediated by SigA are likely associated with the degradation of epithelial fodrin. Consistent with our data, this study also demonstrates that SigA cleaves intracellular fodrin in situ, causing its redistribution within cells. These results strongly implicate SigA in altering the cytoskeleton during the pathogenesis of shigellosis. On the basis of these findings, cleavage of fodrin is a novel mechanism of cellular intoxication for a Shigella toxin. Furthermore, information regarding immunogenicity to SigA in infected patients is lacking. We studied the immune response of SigA from day 28 post-challenge serum of one volunteer from S. flexneri 2a challenge studies. Our results demonstrate that SigA is immunogenic following infection with S. flexneri 2a. CONCLUSIONS: This work shows that SigA binds to epithelial HEp-2 cells as well as being able to induce fodrin degradation in vitro and in situ, further extending its documented role in the pathogenesis of Shigella infections. PMID- 20011052 TI - Long-branch attraction bias and inconsistency in Bayesian phylogenetics. AB - Bayesian inference (BI) of phylogenetic relationships uses the same probabilistic models of evolution as its precursor maximum likelihood (ML), so BI has generally been assumed to share ML's desirable statistical properties, such as largely unbiased inference of topology given an accurate model and increasingly reliable inferences as the amount of data increases. Here we show that BI, unlike ML, is biased in favor of topologies that group long branches together, even when the true model and prior distributions of evolutionary parameters over a group of phylogenies are known. Using experimental simulation studies and numerical and mathematical analyses, we show that this bias becomes more severe as more data are analyzed, causing BI to infer an incorrect tree as the maximum a posteriori phylogeny with asymptotically high support as sequence length approaches infinity. BI's long branch attraction bias is relatively weak when the true model is simple but becomes pronounced when sequence sites evolve heterogeneously, even when this complexity is incorporated in the model. This bias--which is apparent under both controlled simulation conditions and in analyses of empirical sequence data--also makes BI less efficient and less robust to the use of an incorrect evolutionary model than ML. Surprisingly, BI's bias is caused by one of the method's stated advantages--that it incorporates uncertainty about branch lengths by integrating over a distribution of possible values instead of estimating them from the data, as ML does. Our findings suggest that trees inferred using BI should be interpreted with caution and that ML may be a more reliable framework for modern phylogenetic analysis. PMID- 20011054 TI - Effects of Glycyrrhizic Acid on Peroxisome Proliferator-Activated Receptor Gamma (PPARgamma), Lipoprotein Lipase (LPL), Serum Lipid and HOMA-IR in Rats. AB - Studies on ligand binding potential of glycyrrhizic acid, a potential agonist to PPARgamma, displayed encouraging results in amelioration of metabolic syndrome. The regulation of gene cassettes by PPARgamma affects glucose homeostasis, lipid, lipoprotein metabolism and adipogenesis. This study was performed to determine the effects of GA on total PPARgamma and LPL expression levels, lipid parameters and HOMA-IR. Oral administration of 100 mg/kg GA for 24 hours resulted in an increase in insulin sensitivity with decreases in blood glucose, serum insulin and HOMA-IR. Improvement in serum lipid parameters was also observed with a decrease in triacylglycerol, total cholesterol and LDL-cholesterol and an elevation in HDL-cholesterol. GA administration also resulted in up-regulation of total PPARgamma and LPL expression levels in the visceral and subcutaneous adipose tissues, abdominal and quadriceps femoris muscles, as well as liver and kidney, with a significant up-regulation only in the visceral adipose tissue, abdominal and quadriceps femoris muscles. Thus, oral administration of 100 mg/kg GA for 24 hours improved insulin sensitivity and lipid profiles and induced upregulation of total PPARgamma and LPL expression levels in all studied tissues. PMID- 20011053 TI - AtSOFL1 and AtSOFL2 act redundantly as positive modulators of the endogenous content of specific cytokinins in Arabidopsis. AB - BACKGROUND: Although cytokinins have been known for decades to play important roles in the regulation of plant growth and development, our knowledge of the regulatory mechanism of endogenous content of specific cytokinins remains limited. METHODOLOGY/PRINCIPAL FINDINGS: Here, we characterized two SOB five-like (SOFL) genes, AtSOFL1 and AtSOFL2, in Arabidopsis (Arabidopsis thaliana) and showed that they acted redundantly in regulating specific cytokinin levels. Analysis of the translational fusion AtSOFL1:AtSOFL1-GUS and AtSOFL2:AtSOFL2-GUS indicated that AtSOFL1 and AtSOFL2 exhibited similar expression patterns. Both proteins were predominantly expressed in the vascular tissues of developing leaves, flowers and siliques, but barely detectable in roots and stems. Overexpression of either AtSOFL1 or AtSOFL2 led to increased cytokinin content and obvious corresponding mutant phenotypes for both transgenic seedlings and adult plants. In addition, overexpression and site-directed mutagenesis experiments demonstrated that the SOFL domains are necessary for AtSOFL2's overexpression phenotypes. Silencing or disrupting either AtSOFL1 or AtSOFL2 caused no obvious developmental defects. Endogenous cytokinin analysis, however, revealed that compared to the wild type control, the SOFL1-RNAi62 sofl2-1 double mutant accumulated lower levels of trans-zeatin riboside monophosphate (tZRMP) and N(6)-(Delta(2)-isopentenyl)adenosine monophosphate (iPRMP), which are biosynthetic intermediates of bioactive cytokinins. The double mutant also displayed decreased response to exogenous cytokinin in both callus-formation and inhibition-of-hypocotyl-elongation assays. CONCLUSIONS/SIGNIFICANCE: Taken together, our data suggest that in plants AtSOFL1 and AtSOFL2 work redundantly as positive modulators in the fine-tuning of specific cytokinin levels as well as responsiveness. PMID- 20011055 TI - Reactivity of Monovinyl (Meth)Acrylates Containing Cyclic Carbonates. AB - The tremendous diversity of materials properties available with polymers is due in large part to the ability to design structures from the monomeric state. The ease of use of comonomer mixtures only expands this versatility. While final polymer properties are obviously important in the selection or development of a material for a given purpose, for a number of applications, such as optical fiber coatings, photolithography and microelectronics, the additional requirement of a very rapid polymerization process may be equally critical. A class of unusually reactive mono-(meth)acrylate monomers bearing secondary functionality that includes carbonates, carbamates and oxazolidones, has been demonstrated but not fully explained. Here, the influence of an integral cyclic carbonate functional group on (meth)acrylate photopolymerization kinetics is examined in detail with respect to monomers with a wide variety of alternative secondary functionality structure as well as in comparison to conventional mono- and di-(meth)acrylates. The kinetic results from full cure studies of several cyclic carbonate-containing monomers clearly highlight specific structural variations that effectively promote monomer reactivity. Copolymerizations with tetrahydrofurfuryl methacrylate reflect similar dramatic kinetic effects associated with the novel monomers while partial cure homopolymerization studies reveal exceptional dark cure behavior linked to observations of uncommonly low ratios of termination to propagation rates throughout the conversion profile. Temperature effects on reaction kinetics, including both reaction rate and the individual kinetic parameters, as well as the temperature dependence of hydrogen bonding interactions specifically involving the secondary functional groups are probed as a means to understand better the fundamentally interesting and practically important behavior of these monomers. PMID- 20011056 TI - Ectopic calcification as discernible manifestation in neonates with pseudohypoparathyroidism type 1a. AB - The diagnosis of pseudohypoparathyroidism type 1a (PHP1a) is challenging, because both the osteodystrophy, such as brachydactyly and round face, and the symptomatic hypocalcemia usually develop beyond infancy. Although ectopic calcification may be an early sign of PHP1a, there are no systematic reviews regarding the time of its appearance. We here report on two PHP1a patients who presented with subcutaneous calcification in neonatal period. PMID- 20011057 TI - Cholecystokinin and Somatostatin Negatively Affect Growth of the Somatostatin-RIN 14B Cells. AB - With the exclusive presence of the pancreatic CCK-2 receptors on the pancreatic delta cells of six different species, this study was undertaken to determine the role of cholecystokinin and gastrin on growth of these somatostatin (SS) cells. For this study, the SS-RIN-14B cells were used in culture and their growth was evaluated by cell counting. Results. To our surprise, we established by Western blot that these RIN cells possess the two CCK receptor subtypes, CCK-1 and CCK-2. Occupation of the CCK-1 receptors by caerulein, a CCK analog, led to inhibition of cell proliferation, an effect prevented by a specific CCK-1 receptor antagonist. Occupation of the CCK-2 receptors by the gastrin agonist pentagastrin had no effect on cell growth. Proliferation was not affected by SS released from these cells but was inhibited by exogenous SS. Conclusions. Growth of the SS-RIN 14B cells can be negatively affected by occupation of their CCK-1 receptors and by exogenous somatostatin. PMID- 20011058 TI - Brown tumour in a patient with secondary hyperparathyroidism resistant to medical therapy: case report on successful treatment after subtotal parathyroidectomy. AB - Brown tumour represents a serious complication of hyperparathyroidism. Differential diagnosis, based on histological examination, is only presumptive and clinical, radiological and laboratory data are necessary for definitive diagnosis. Here we describe a case of a brown tumour localised in the maxilla due to secondary hyperparathyroidism in a young women with chronic renal failure. Hemodialysis and pharmacological treatment were unsuccessful in controlling secondary hyperparathyroidism making it necessary to proceed with a subtotal parathyroidectomy. The proper timing of the parathyroidectomy and its favourable effect on regression of the brown tumor made it possible to avoid a potentially disfiguring surgical removal of the brown tumor. PMID- 20011059 TI - Trends in Method of Diagnosis of Type 2 Diabetes Mellitus: Results from SHIELD. AB - Aims. This study assessed whether recent screening recommendations have led to increased diagnosis of type 2 diabetes mellitus (T2DM) through routine screening. Methods. Respondents to the 2006 US SHIELD survey reported whether a physician told them they had T2DM, age at diagnosis, specialty of the physician who made the diagnosis, and whether the diagnosis was made after having symptoms, during routine screening, or when being treated for another health problem. Results. Of 3 022 T2DM respondents, 36% of respondents reported that T2DM diagnosis was made during routine screening alone, 20% after having symptoms alone, and 6% when being treated for another health problem alone. The proportion of T2DM respondents reporting a diagnosis based only on screening increased approximately 42% over a 15+-year time span (absolute increase from 31% to 44%) (P < .001), whereas symptom-based diagnosis did not change significantly (P = .10). T2DM was diagnosed primarily by family physicians (88.3%). Conclusion. These findings highlight the importance of regular screening for diabetes and the vital role of primary care physicians in recognizing individuals with T2DM. PMID- 20011060 TI - Prolactin and male fertility: the long and short feedback regulation. AB - In the last 20 years, a pituitary-hypothalamus tissue culture system with intact neural and portal connections has been developed in our lab and used to understand the feedback mechanisms that regulate the secretions of adenohypophyseal hormones and fertility of male rats. In the last decade, several in vivo rat models have also been developed in our lab with a view to substantiate the in vitro findings, in order to delineate the role of pituitary hormones in the regulation of fertility of male rats. These studies have relied on both surgical and pharmacological interventions to modulate the secretions of gonadotropins and testosterone. The interrelationship between the circadian release of reproductive hormones has also been ascertained in normal men. Our studies suggest that testosterone regulates the secretion of prolactin through a long feedback mechanism, which appears to have been conserved from rats to humans. These studies have filled in a major lacuna pertaining to the role of prolactin in male reproductive physiology by demonstrating the interdependence between testosterone and prolactin. Systemic levels of prolactin play a deterministic role in the mechanism of chromatin condensation during spermiogenesis. PMID- 20011062 TI - The contribution of three-dimensional power Doppler imaging in the preoperative assessment of breast tumors: a preliminary report. AB - Purpose. The aim of this study was to determine the value of 3D and 3D Power Doppler sonography in the detection of tumor malignancy in breast lesions and to find new diagnostic criteria for differential diagnosis. Methods. One hundred and twenty five women with clinically or mammographically suspicious findings were referred for 3D Power Doppler ultrasound prior to surgery. Histological diagnosis was conducted after surgery and compared with ultrasound findings. Sonographic criteria used for breast cancer diagnosis were based on a system that included morphological characteristics and criteria of the vascular pattern of a breast mass by Power Doppler imaging. Results. Seventy-two lesions were histopathologically diagnosed as benign and 53 tumors as malignant. Three dimensional ultrasound identified 49 out of 53 histologically confirmed breast cancers resulting in a sensitivity of 92.4% and a specificity of 86.1% in diagnosing breast malignancy (PPV: 0.83, NPV:0.94). Conclusions. 3D ultrasonography is a valuable tool in identifying preoperatively the possibility of a tumor to be malignant. PMID- 20011061 TI - The potential use of intrauterine insemination as a basic option for infertility: a review for technology-limited medical settings. AB - Objective. There is an asymmetric allocation of technology and other resources for infertility services. Intrauterine insemination (IUI) is a process of placing washed spermatozoa transcervically into the uterine cavity for treatment of infertility. This is a review of literature for the potential use of IUI as a basic infertility treatment in technology-limited settings. Study design. Review of articles on treatment of infertility using IUI. Results. Aspects regarding the use of IUI are reviewed, including ovarian stimulation, semen parameters associated with good outcomes, methods of sperm preparation, timing of IUI, and number of inseminations. Implications of the finding in light of the needs of low technology medical settings are summarized. Conclusion. The reviewed evidence suggests that IUI is less expensive, less invasive, and comparably effective for selected patients as a first-line treatment for couples with unexplained or male factor infertility. Those couples may be offered three to six IUI cycles in technology-limited settings. PMID- 20011063 TI - Regional brain stem atrophy in idiopathic Parkinson's disease detected by anatomical MRI. AB - Idiopathic Parkinson's disease (PD) is a neurodegenerative disorder characterized by the dysfunction of dopaminergic dependent cortico-basal ganglia loops and diagnosed on the basis of motor symptoms (tremors and/or rigidity and bradykinesia). Post-mortem studies tend to show that the destruction of dopaminergic neurons in the substantia nigra constitutes an intermediate step in a broader neurodegenerative process rather than a unique feature of Parkinson's disease, as a consistent pattern of progression would exist, originating from the medulla oblongata/pontine tegmentum. To date, neuroimaging techniques have been unable to characterize the pre-symptomatic stages of PD. However, if such a regular neurodegenerative pattern were to exist, consistent damages would be found in the brain stem, even at early stages of the disease. We recruited 23 PD patients at Hoenn and Yahr stages I to II of the disease and 18 healthy controls (HC) matched for age. T1-weighted anatomical scans were acquired (MPRAGE, 1 mm3 resolution) and analyzed using an optimized VBM protocol to detect white and grey matter volume reduction without spatial a priori. When the HC group was compared to the PD group, a single cluster exhibited statistical difference (p<0.05 corrected for false detection rate, 4287 mm3) in the brain stem, between the pons and the medulla oblongata. The present study provides in-vivo evidence that brain stem damage may be the first identifiable stage of PD neuropathology, and that the identification of this consistent damage along with other factors could help with earlier diagnosis in the future. This damage could also explain some non motor symptoms in PD that often precede diagnosis, such as autonomic dysfunction and sleep disorders. PMID- 20011064 TI - Excreted/secreted proteins from trypanosome procyclic strains. AB - Trypanosoma secretome was shown to be involved in parasite virulence and is suspected of interfering in parasite life-cycle steps such as establishment in the Glossina midgut, metacyclogenesis. Therefore, we attempted to identify the proteins secreted by procyclic strains of T. brucei gambiense and T. brucei brucei, responsible for human and animal trypanosomiasis, respectively. Using mass spectrometry, 427 and 483 nonredundant proteins were characterized in T. brucei brucei and T. brucei gambiense secretomes, respectively; 35% and 42% of the corresponding secretome proteins were specifically secreted by T. brucei brucei and T. brucei gambiense, respectively, while 279 proteins were common to both subspecies. The proteins were assigned to 12 functional classes. Special attention was paid to the most abundant proteases (14 families) because of their potential implication in the infection process and nutrient supply. The presence of proteins usually secreted via an exosome pathway suggests that this type of process is involved in trypanosome ESP secretion. The overall results provide leads for further research to develop novel tools for blocking trypanosome transmission. PMID- 20011066 TI - Infection with Hymenolepis diminuta is more effective than daily corticosteroids in blocking chemically induced colitis in mice. AB - PURPOSE: To compare infection with the tapeworm, Hymenolepis diminuta, with steroid (dexamethasone) administration in the inhibition of dinitrobenzene sulfonic acid- (DNBS-) induced colitis in mice. PROCEDURES: Mice were treated with DNBS +/- infected with H. diminuta or treated with daily dexamethasone (2 mg/Kg, ip.) and were assessed 72 hours post-DNBS by the calculation of disease activity and histological damage scores, and spleen cell cytokine production. RESULTS: H. diminuta-infected mice showed increased IL-4 and IL-10 production by spleen cells compared to other groups and were protected from DNBS-induced colitis. In contrast, there was little benefit of dexamethasone in the treatment of colitis. Collagen deposition in the colon was not different between the groups. CONCLUSIONS: H. diminuta was superior to dexamethasone in the prevention of DNBS-induced colitis and did not result in additional side effects (i.e., collagen deposition). Comparisons with current therapeutics and long-term followup to studies are essential if "helminth therapy" is to become a viable treatment for specific inflammatory diseases in the gut or other tissues. PMID- 20011065 TI - Mechanisms of the regulation of the intestinal Na+/H+ exchanger NHE3. AB - A major of Na(+) absorptive process in the proximal part of intestine and kidney is electroneutral exchange of Na(+) and H(+) by Na(+)/H(+) exchanger type 3 (NHE3). During the past decade, significant advance has been achieved in the mechanisms of NHE3 regulation. A bulk of the current knowledge on Na(+)/H(+) exchanger regulation is based on heterologous expression of mammalian Na(+)/H(+) exchangers in Na(+)/H(+) exchanger deficient fibroblasts, renal epithelial, and intestinal epithelial cells. Based on the reductionist's approach, an understanding of NHE3 regulation has been greatly advanced. More recently, confirmations of in vitro studies have been made using animals deficient in one or more proteins but in some cases unexpected findings have emerged. The purpose of this paper is to provide a brief overview of recent progress in the regulation and functions of NHE3 present in the luminal membrane of the intestinal tract. PMID- 20011067 TI - Molecular mechanisms of receptor-mediated endocytosis in the renal proximal tubular epithelium. AB - Receptor-mediated endocytosis is a pivotal function of renal proximal tubule epithelial cells (PTECs) to reabsorb and metabolize substantial amounts of proteins and other substances in glomerular filtrates. The function accounts for the conservation of nutrients, including carrier-bound vitamins and trace elements, filtered by glomeruli. Impairment of the process results in a loss of such substances and development of proteinuria, an important clinical sign of kidney disease and a risk marker for cardiovascular disease. Megalin is a multiligand endocytic receptor expressed at clathrin-coated pits of PTEC, playing a central role in the process. Megalin cooperates with various membrane molecules and interacts with many intracellular adaptor proteins for endocytic trafficking. Megalin is also involved in signaling pathways in the cells. Megalin-mediated endocytic overload leads to damage of PTEC. Further studies are needed to elucidate the mechanism of megalin-mediated endocytosis and develop strategies for preventing the damage of PTEC. PMID- 20011068 TI - Genome-wide identification and evolutionary analysis of the animal specific ETS transcription factor family. AB - The ETS proteins are a family of transcription factors (TFs) that regulate a variety of biological processes. We made genome-wide analyses to explore the classification of the ETS gene family. We identified 207 ETS genes which encode 321 ETS TFs from ten animal species. Of the 321 ETS TFs, 155 contain only an ETS domain, about 50% contain a ETS_PEA3_N or a SAM_PNT domain in addition to an ETS domain, the rest (only four) contain a second ETS domain or a second ETS_PEA3_N domain or an another domain (AT_hook or DNA_pol_B). A Neighbor-Joining phylogenetic tree was constructed using the amino acid sequences of the ETS domain of the ETS TFs. The results revealed that the ETS genes of the ten species can be divided into two distinct groups. Group I contains one nematode ETS gene and 18 vertebrate animal ETS genes. Group II contains the majority of the ETS TFs and can be further divided into eleven subgroups. The sequence motifs outside the DNA-binding domain and the conservation of the exon-intron structural patterns of the ETS TFs in human, cattle, and chicken further support the phylogenetic classification among these ETS TFs. Extensive duplication of the ETS genes was found in the genome of each species. The duplicated ETS genes account for ~69% of the total of ETS genes. Furthermore, we also found there are ETS gene clusters in all of the ten animal species. Statistical analysis of the Gene Ontology annotations of the ETS genes showed that the ETS proteins tend to be related to RNA biosynthetic process, biopolymer metabolic process and macromolecule metabolic process expected from the common GO categories of transcriptional factors. We also discussed the functional conservation and diversification of ETS TFs. PMID- 20011069 TI - Detection of EBV infection and gene expression in oral cancer from patients in Taiwan by microarray analysis. AB - Epstein-Barr virus is known to cause nasopharyngeal carcinoma. Although oral cavity is located close to the nasal pharynx, the pathogenetic role of Epstein Barr virus (EBV) in oral cancers is unclear. This molecular epidemiology study uses EBV genomic microarray (EBV-chip) to simultaneously detect the prevalent rate and viral gene expression patterns in 57 oral squamous cell carcinoma biopsies (OSCC) collected from patients in Taiwan. The majority of the specimens (82.5%) were EBV-positive that probably expressed coincidently the genes for EBNAs, LMP2A and 2B, and certain structural proteins. Importantly, the genes fabricated at the spots 61 (BBRF1, BBRF2, and BBRF3) and 68 (BDLF4 and BDRF1) on EBV-chip were actively expressed in a significantly greater number of OSCC exhibiting exophytic morphology or ulceration than those tissues with deep invasive lesions (P = .0265 and .0141, resp.). The results may thus provide the lead information for understanding the role of EBV in oral cancer pathogenesis. PMID- 20011070 TI - Differential midgut attachment of Leishmania (Viannia) braziliensis in the sand flies Lutzomyia (Nyssomyia) whitmani and Lutzomyia (Nyssomyia) intermedia. AB - The interaction between Leishmania and sand flies has been demonstrated in many Old and New World species. Besides the morphological differentiation from procyclic to infective metacyclic promastigotes, the parasite undergoes biochemical transformations in its major surface lipophosphoglycan (LPG). An upregulation of beta-glucose residues was previously shown in the LPG repeat units from procyclic to metacyclic phase in Leishmania (Viannia) braziliensis, which has not been reported in any Leishmania species. LPG has been implicated as an adhesion molecule that mediates the interaction with the midgut epithelium of the sand fly in the Subgenus Leishmania. These adaptations were explored for the first time in a species from the Subgenus Viannia, L. (V.) braziliensis with its natural vectors Lutzomyia (Nyssomyia) intermedia and Lutzomyia (Nyssomyia) whitmani. Using two in vitro binding techniques, phosphoglycans (PGs) derived from procyclic and metacyclic parasites were able to bind to the insect midgut and inhibit L. braziliensis attachment. Interestingly, L. braziliensis procyclic parasite attachment was approximately 11-fold greater in the midgut of L. whitmani than in L. intermedia. The epidemiological relevance of L. whitmani as a vector of American Cutaneous Leishmaniasis (ACL) in Brazil is discussed. PMID- 20011071 TI - Endocytosis and recycling of tight junction proteins in inflammation. AB - A critical function of the epithelial lining is to form a barrier that separates luminal contents from the underlying interstitium. This barrier function is primarily regulated by the apical junctional complex (AJC) consisting of tight junctions (TJs) and adherens junctions (AJs) and is compromised under inflammatory conditions. In intestinal epithelial cells, proinflammatory cytokines, for example, interferon-gamma (IFN-gamma), induce internalization of TJ proteins by endocytosis. Endocytosed TJ proteins are passed into early and recycling endosomes, suggesting the involvement of recycling of internalized TJ proteins. This review summarizes mechanisms by which TJ proteins under inflammatory conditions are internalized in intestinal epithelial cells and point out comparable mechanism in nonintestinal epithelial cells. PMID- 20011073 TI - The Role of Advocacy Organizations in Reducing Negative Externalities. AB - An externality is a cost that a corporation's actions impose on society. For example, a power plant may emit mercury, but might not pay for the cost of that pollution to the people living near the plant. It is possible to analyze a diverse range of problems of society in these terms, including the health effects of corporate practices, the unsustainability of manufacturing processes, and marketing of products contributing to environmental damage, and economic policies that maintain high levels of poverty due to effective lobbying by the business community. This paper examines the problem of externalities in terms of metacontingencies. Externalities continue precisely because there is no cost to the organizations for practices that impose these costs on third parties. The paper describes the cultural practices needed to influence governments are motivated to make corporations bear the true costs of their practices-costs that are currently imposed on others. PMID- 20011072 TI - The mysterious unfoldome: structureless, underappreciated, yet vital part of any given proteome. AB - Contrarily to the general believe, many biologically active proteins lack stable tertiary and/or secondary structure under physiological conditions in vitro. These intrinsically disordered proteins (IDPs) are highly abundant in nature and many of them are associated with various human diseases. The functional repertoire of IDPs complements the functions of ordered proteins. Since IDPs constitute a significant portion of any given proteome, they can be combined in an unfoldome; which is a portion of the proteome including all IDPs (also known as natively unfolded proteins, therefore, unfoldome), and describing their functions, structures, interactions, evolution, and so forth. Amino acid sequence and compositions of IDPs are very different from those of ordered proteins, making possible reliable identification of IDPs at the proteome level by various computational means. Furthermore, IDPs possess a number of unique structural properties and are characterized by a peculiar conformational behavior, including their high stability against low pH and high temperature and their structural indifference toward the unfolding by strong denaturants. These peculiarities were shown to be useful for elaboration of the experimental techniques for the large scale identification of IDPs in various organisms. Some of the computational and experimental tools for the unfoldome discovery are discussed in this review. PMID- 20011074 TI - Trisomy 21 in one of extremely low birth weight twins. AB - Prematurity is frequently seen in the neonatal intensive care unit, and trisomy 21 is an often diagnosed neonatal disorder. We report a unique case of extremely premature twins, one of whom was ultimately diagnosed with trisomy 21. We were able to examine the neonatal courses and outcomes of these twins, which were similar despite the presence of trisomy 21 in one twin. This is the first report comparing the neonatal course of an infant with trisomy 21 to an unaffected twin in patients born so prematurely, and demonstrates the difficulty of making the diagnosis of trisomy 21 based solely on physical examination in premature infants. PMID- 20011075 TI - Development of an Automated Steering Mechanism for Bladder Urothelium Surveillance. AB - Given the advantages of cystoscopic exams compared with other procedures available for bladder surveillance, it would be beneficial to develop an improved automated cystoscope. We develop and propose an active programmable remote steering mechanism and an efficient motion sequence for bladder cancer detection and postoperative surveillance. The continuous and optimal path of the imaging probe can enable a medical practitioner to readily ensure that images are produced for the entire surface of the bladder in a controlled and uniform manner. Shape memory alloy (SMA) based segmented actuators disposed adjacent to the distal end of the imaging probe are selectively activated to bend the shaft to assist in positioning and orienting the imaging probe at a plurality of points selected to image all the interior of the distended bladder volume. The bending arc, insertion depth, and rotational position of the imaging probe are automatically controlled based on patient-specific data. The initial prototype is tested on a 3D plastic phantom bladder, which is used as a proof-of-concept in vitro model and an electromagnetic motion tracker. The 3D tracked tip trajectory results ensure that the motion sequencing program and the steering mechanism efficiently move the image probe to scan the entire inner tissue layer of the bladder. The compared experimental results shows 5.1% tip positioning error to the designed trajectory given by the simulation tool. The authors believe that further development of this concept will help guarantee that a tumor or other characteristic of the bladder surface is not overlooked during the automated cystoscopic procedure due to a failure to image it. PMID- 20011076 TI - Competition Between Transients in the Rate of Approach to a Fixed Point. AB - The goal of this paper is to provide and apply tools for analyzing a specific aspect of transient dynamics not covered by previous theory. The question we address is whether one component of a perturbed solution to a system of differential equations can overtake the corresponding component of a reference solution as both converge to a stable node at the origin, given that the perturbed solution was initially farther away and that both solutions are nonnegative for all time. We call this phenomenon tolerance, for its relation to a biological effect. We show using geometric arguments that tolerance will exist in generic linear systems with a complete set of eigenvectors and in excitable nonlinear systems. We also define a notion of inhibition that may constrain the regions in phase space where the possibility of tolerance arises in general systems. However, these general existence theorems do not not yield an assessment of tolerance for specific initial conditions. To address that issue, we develop some analytical tools for determining if particular perturbed and reference solution initial conditions will exhibit tolerance. PMID- 20011077 TI - Effects of triamcinolone acetonide on vessels of the posterior segment of the eye. AB - PURPOSE: This study investigates the effects of triamcinolone acetonide (TA) on retinal endothelial cells in vitro and explores the potential vascular toxic effect of TA injected into the vitreous cavity of rats in vivo. METHODS: Subconfluent endothelial cells were treated with either 0.1 mg/ml or 1 mg/ml TA in 1% ethanol. Control cells were either untreated or exposed to 1% ethanol. Cell viability was evaluated at 24 h, 72 h, and five days using the tetrazolium 3-(4,5 dimethylthiazol-2-yl)-2,5 phenyltetrazolium bromide test (MTT) and lactate dehydrogenase (LDH) assays. Cell proliferation was evaluated by 5-bromo-2 deoxyuridine (BrdU) test. Apoptosis was evaluated by terminal deoxynucleotidyl transferase dUTP nick end labeling assay (TUNEL assay), annexin-binding, and caspase 3 activation. Caspase-independent cell deaths were investigated by immunohistochemistry using antibodies against apoptosis inducing factor (AIF), cytochrome C, microtubule-associated protein (MAP)-light chain 3 (MAP-LC3), and Leukocyte Elastase Inhibitor/Leukocyte Elastase Inhibitor-derived DNase II (LEI/L DNase II). In vivo, semithin and ultrathin structure analysis and vascular casts were performed to examine TA-induced changes of the choroidal vasculature. In addition, outer segments phagocytosis assay on primary retinal pigment epithelium (RPE) cells was performed to assess cyclooxygenase (COX-2) and vascular endothelial growth factor (VEGF) mRNAs upregulation with or without TA. RESULTS: The inhibitory effect of TA on cell proliferation could not explain the significant reduction in cell viability. Indeed, TA induced a time-dependent reduction of bovine retinal endothelial cells viability. Annexin-binding positive cells were observed. Cytochrome C was not released from mitochondria. L-DNase II was found translocated to the nucleus, meaning that LEI was changed into L-DNase II. AIF was found nuclearized in some cells. LC3 labeling showed the absence of autophagic vesicles. No autophagy or caspase dependent apoptosis was identified. At 1 mg/ml TA induced necrosis while exposure to lower concentrations for 3 to 5 days induced caspase independent apoptosis involving AIF and LEI/L-DNase II. In vivo, semithin and ultrathin structure analysis and vascular casts revealed that TA mostly affected the choroidal vasculature with a reduction of choroidal thickness and increased the avascular areas of the choriocapillaries. Experiments performed on primary RPE cells showed that TA downregulates the basal expression of COX-2 and VEGF and inhibits the outer segments (OS)-dependent COX-2 induction but not the OS-dependent VEGF induction. CONCLUSIONS: This study demonstrates for the first time that glucocorticoids exert direct toxic effect on endothelial cells through caspase-independent cell death mechanisms. The choroidal changes observed after TA intravitreous injection may have important implications regarding the safety profile of TA use in human eyes. PMID- 20011078 TI - Gradients of Eph-A6 expression in primate retina suggest roles in both vascular and axon guidance. AB - PURPOSE: Recently we identified high levels of expression of Eph-A6 in the macula of developing human retina and showed localization of Eph-A6 to ganglion cells (GC). In the present study we investigated the expression of some members of the ephrin family in developing primate retina, including the topography of Eph-A6 expression, and its ligands, in developing macaque retinas. METHODS: We extracted RNA from human fetal retinas and probed for Eph-A5-A7, Eph-B1, ephrin-B2, and ephrin-A1-A5 by RT-PCR, then prepared riboprobes for Eph-A5-A7, Eph-B1 and ephrin A1, -A4 and -B2. Paraffin sections of fetal macaque retinas were used to localize expression of Ephs and ephrins by in situ hybridization and immunohistochemistry. RESULTS: We identified prominent gradients of Eph-A6 mRNA expression in the ganglion cell layer (GCL) of fetal macaque retinas of different ages. The gradient of Eph-A6 expression was high near the optic disc and low at the developing macula at fetal day (Fd) 55. At Fd 70 and 80, the gradient of Eph-A6 expression was reversed, being higher temporal to the macula, and low at the disc. By Fd 110, when the fovea begins to form, a pattern of expression was established that persisted into the postnatal period, in which the highest levels of expression were detected at the developing fovea, and progressively lower levels of expression were detected at increasing distance from the fovea. Beginning at Fd 70, we also detected a gradient of Eph-A6 expression running perpendicular to the retinal surface within the GCL of central retina that was high in the inner GCL and low in the outer GCL. This second pattern persisted into the neonatal period. We found the two ligands for Eph-A6, ephrin-A1 and ephrin-A4, expressed by Pax2-immunoreactive astrocytes, in the optic nerve head and in the retina, by in situ hybridization and immunohistochemistry. We propose that during development of the retinal vasculature, migration of ligand-bearing astrocytes is slowed along this Eph-A6 expression gradient through repellent Eph A6 - ephrin-A1 and -A4 signaling. CONCLUSIONS: Patterns of Eph-A6 expression in the developing macaque retina suggest that Eph-A6 - ephrin-A1 and -A4 repellent signaling has a role in retinal vascular patterning, and in the postnatal maintenance of projections from macular and foveal GC. PMID- 20011080 TI - PKC alpha affects cell cycle progression and proliferation in human RPE cells through the downregulation of p27kip1. AB - PURPOSE: Protein kinase C (PKC) plays an important role in the regulation of retinal pigment epithelium (RPE) cell proliferation. In this study, we investigated which of these isozymes could be responsible for the cell cycle and proliferation in human RPE cells. METHODS: The effect of PKC activators on human RPE cell cycle progression was tested by flow cytometry. To identify the isoform of PKC responsible for the increased progression of the cells through the cell cycle, we monitored the effect of phorbol 12-myristate 13-acetate (PMA) on the subcellular localization of the nine PKC isoforms expressed in RPE cells. To evaluate the molecular mechanism by which PKC(alpha) induces cell cycle progression, we examined the transcript, protein, and cellular levels of cell cycle regulatory proteins using RT-PCR, western blotting, and a confocal microscope, respectively. RESULTS: We demonstrated that PKC activation by PMA affected cell cycle progression in RPE cells. Of the nine PKC isoforms that were present in RPE cells, we found PKC(alpha) was both necessary and sufficient to promote cell cycle progression after being stimulated with PMA. Decreased PKC(alpha) expression resulted in a significant decrease in cell proliferation. The only cell cycle-regulatory molecule whose expression was rapidly altered and decreased by PKC(alpha) activity was the cyclin- dependent kinase (CDK) inhibitor p27(kip1). CONCLUSIONS: These results suggest that PKC(alpha) affects cell cycle progression and proliferation in human RPE cells through the downregulation of p27(kip1). PMID- 20011079 TI - Lack of association between toll-like receptor 4 gene polymorphisms and sarcoidosis-related uveitis in Japan. AB - PURPOSE: Toll-like receptors (TLRs) are pattern-recognition receptors that play an important role in innate and adaptive immune responses to microbial pathogens. Among TLRs, TLR4 recognizes lipopolysaccharides of Gram-negative bacteria. Genetic polymorphisms within the TLR4 gene have been reported to be associated with various inflammatory diseases; therefore, TLR4 appears to be a susceptibility gene for sarcoidosis. Although sarcoidosis has various clinical manifestations, its association with uveitis is more common in Japan than in other countries. The aim of this study was to investigate whether TLR4 polymorphisms were associated with sarcoidosis-related uveitis in a Japanese population. METHODS: Two hundred twenty-three patients with sarcoidosis and 206 healthy control subjects were recruited at seven sites in Japan. Eight single nucleotide polymorphisms (SNPs) in TLR4 were genotyped with a TaqMan assay, and allelic and phenotypic diversity were assessed in affected and control subjects. RESULTS: We found no association with susceptibility to sarcoid-related uveitis for any of the SNPs analyzed. Strong linkage disequilibrium was observed among all the SNPs analyzed (D'>/=0.78), which were located in one haplotype block. CONCLUSION: TLR4 polymorphisms do not play an important role in the development of uveitis in Japanese patients with sarcoidosis. PMID- 20011081 TI - Extracellular SOD and VEGF are increased in vitreous bodies from proliferative diabetic retinopathy patients. AB - PURPOSE: To evaluate the relationship between vascular endothelial growth factor (VEGF) and extracellular superoxide dismutase (EC-SOD) in vitreous body and serum in patients with proliferative diabetic retinopathy (PDR), and investigate the role of EC-SOD in PDR by evaluating its angiostatic effect, using an in vitro angiogenesis model. To investigate the role of EC-SOD in PDR by evaluating its angiostatic effect, using an in vitro angiogenesis model. METHODS: EC-SOD and VEGF concentrations in vitreous and serum samples from PDR and macular hole (MH) were measured by ELISA. The effects of EC-SOD on VEGF-induced proliferation, migration, and tube formation were evaluated using human umbilical vein endothelial cells (HUVECs). Moreover, the effects of EC-SOD on VEGF-induced proliferation and migration were evaluated in HUVECs and primary normal human retinal microvascular endothelial cells. RESULTS: Intravitreal concentrations of EC-SOD were significantly higher (p<0.01) in PDR (58.0+/-23.8 ng/ml, mean+/-SD) than in MH (29.3+/-6.6 ng/ml). Intravitreal concentrations of VEGF were dramatically higher (p<0.01) in PDR (798.2+/-882.7 pg/ml) than in MH (17.7+/-15.5 pg/ml). The serum concentrations of EC-SOD and VEGF did not differ between the two patient groups. The vitreous concentrations of VEGF correlated with those of EC-SOD in all patients (rs=0.61, p<0.001). In HUVECs, EC-SOD at 100 ng/ml significantly suppressed VEGF-induced proliferation and tube formation, but not VEGF-induced migration. CONCLUSIONS: EC-SOD was increased together with VEGF in the vitreous body from PDR patients, suggesting that EC-SOD may play a pivotal role in the pathogenesis of angiogenesis. PMID- 20011082 TI - The diabetic foot: a global threat and a huge challenge for Greece. AB - The diabetic foot continues to be a major cause of morbidity, posing a global threat. Substantial progress has been now accomplished in the treatment of foot lesions, but further improvement is required. Treatment options may be classified into established measures (revascularisation, casting and debridement) and new modalities. All therapeutic measures should be provided by specialised dedicated multidisciplinary foot clinics. In particular, the diabetic foot is a huge challenge for Greece. There is a dramatic need to increase the number of engaged foot care teams and their resources throughout the country. It is also desirable to continue education of both physicians and general diabetic population on the magnitude of the problem and on the suitable preventative measures. At the same time, more data on the prevalence and clinical manifestations of the diabetic foot in Greece should be carefully collected. Finally, additional research should investigate feasible ways of implementing current knowledge in everyday clinical practice. PMID- 20011083 TI - Brain dead donor kidneys are immunologically active: is intervention justified? AB - The improvement in the field of kidney transplantation, during the last decades, has brought kindey transplantation to the top of patient preference as the best kidney replacement therapy. The use of marginal kidney grafts, which are highly immunogenic has become common practice because of lack of kidney donors. Inflammatory activity in the kidneys after brain death is an ongoing phenomenon. The inappropriate treatment of brain dead donor may result to primary non function (PNF) of the graft, delayed graft function (DGF) or to long term graft dysfunction and shortened graft survival. Therefore correct handling of the brain dead donor is of paramount importance. The impact of various pharmacologic agents (catecholamines, glucocorticoids, carbamylated recombinant human erythropoietin, recombinant soluble P-selectin glycoprotein ligant, heme oxygenase-1, carbon monoxide, and mycophenolate mofetil) on the immunogenicity of brain dead donor kidneys is discussed. PMID- 20011084 TI - Hepatitis C and liver transplantation. AB - Cirrhosis due to chronic hepatitis C is the leading indication for liver transplantation in Europe, United States and Japan. Reinfection after liver transplantation is universal and chronic liver disease develops in at least 70% of patients at 3 years, with an accelerated course compared to the nontransplant setting. These facts underscore the need for a better understanding of hepatitis C infection and the various treatment modalities. This paper attempts a brief review of the scope of the disease, as well as the different treatment modalities, with special emphasis given to orthotopic liver transplantation. PMID- 20011086 TI - Renal fibrosis. AB - Tubulointerstitial renal fibrosis, characterized as a progressive detrimental connective tissue deposition on the kidney parenchyma, appears to be a harmful process leading inevitably to renal function deterioration, independently of the primary renal disease which causes the original kidney injury. Epithelial to Mesenchymal Transition (EMT) of tubular epithelial cells which are transformed to mesenchymal fibroblasts migrating to adjacent interstitial parenchyma constitutes the principal mechanism of renal fibrosis along with local and circulating cells. Proteinuria as well as hypoxia is included among the main mechanisms of EMT stimulation. TGFbeta-1 through the SMAD pathway is considered as the main modulator regulating the EMT molecular mechanism, probably in cooperation with hypoxia inducible factors. Hepatocyte Growth Factor (HGF) and Bone Morphogenetic Factor-7 (BMF-7) are inhibitory to EMT molecules which could prevent in experimental and clinical level the catastrophic process of interstitial fibrosis. Interesting data emerge indicating that HGF and BMF-7 administration prevents the peritoneal fibrosis of mesothelial cells. PMID- 20011087 TI - Impact of computer use on children's vision. AB - Today, millions of children use computers on a daily basis. Extensive viewing of the computer screen can lead to eye discomfort, fatigue, blurred vision and headaches, dry eyes and other symptoms of eyestrain. These symptoms may be caused by poor lighting, glare, an improper work station set-up, vision problems of which the person was not previously aware, or a combination of these factors. Children can experience many of the same symptoms related to computer use as adults. However, some unique aspects of how children use computers may make them more susceptible than adults to the development of these problems. In this study, the most common eye symptoms related to computer use in childhood, the possible causes and ways to avoid them are reviewed. PMID- 20011085 TI - Genetics of polycystic ovary syndrome. AB - Polycystic ovary syndrome (PCOS) is a syndrome involving defects in primary cellular control mechanisms that result in the expression of chronic anovulation and hyperandrogenism. This syndrome has been for many years one of the most controversial entities in gynecological endocrinology. Polycystic ovary syndrome has been proven to be a familial condition. Although the role of genetic factors in PCOS is strongly supported, the genes that are involved in the etiology of the syndrome have not been fully investigated until now, as well as the environmental contribution in their expression. The heterogeneity of the syndrome entertains the mystery around this condition which concerns thousands of infertile women worldwide. Some genes have shown altered expression suggesting that the genetic abnormality in PCOS affects signal transduction pathways controlling steroidogenesis, steroid hormones action, gonadotrophin action and regulation, insulin action and secretion, energy homeostasis, chronic inflammation and others. The present review of the contemporary literature constitutes an effort to present all the trends in the current research for the etiology of polycystic ovary syndrome. PMID- 20011088 TI - Does prostate specific antigen density correlates with aggressiveness of the prostate cancer? AB - BACKGROUND: As already documented, a high prostate specific antigen in men with normal size of prostate gland is more likely to be associated with an aggressive cancer as compared to others with the same prostate specific antigen and a large gland size. In this retrospective study we tested the association between Prostate Specific Antigen Density (PSAD) and tumor aggressiveness in patients with clinically localized Prostate Cancer (PCa) surgically treated by radical prostatectomy. METHODS: We evaluated data from patients records in a cohort of 72 patients who underwent radical prostatectomy between January 2000 and June 2007. PSAD was calculated as ratio between the preoperative total prostatic specific antigen (PSA) in nanograms per milliliter with the prostate weight (PW) of prostatectomized specimen in grams or prostate volume measured with ultrasound (US). The patients were stratified into four PSAD categories: 0.1-0.15, 0.16- 0.20, 0.21-0.5 and greater than 0.51 ng/ml/gr. Parameters that were included into analysis were: PSA, measurement of the prostate volume by ultrasound (preoperatively) and prostate weight, pathological tumor stage, Gleason sum, Gleason grade, metastatic lymph nodes, seminal vesicle involvement and organ confine disease (postoperatively). Worsening of the clinicopathological properties was defined as aggressiveness. RESULTS: There was a significant correlation between US-PSAD and PW-PSAD (p<0.001). In US-PSAD categories the statistic tests found significant correlation with the primary tumor (R=0.303, p<0.01), metastatic lymph nodes (R=0.331, p<0.01), and the organ confine disease (R=0.296, p<0.05). The PW-PSAD categories correlated significantly with the pathologic findings from other parameters. Hence, a statistically significant correlation was found with Gleason sum (R=0.246, p<0.05), Gleason grade (R=0.234, p<0.05), primary tumor (R=0.285, p<0.05), metastatic lymph node (R=0.287, p<0.05) and organ confine disease (R=0.303, p<0.01). CONCLUSIONS: Prostate specific antigen density measurement is useful tool for the assessment of the degree of aggressiveness in clinically localized prostate cancer, and further investigation regarding its possible use as a prediction marker is justified. PMID- 20011089 TI - Coagulation factors as biological risk markers of endothelial dysfunction. Association with the thrombotic episodes of chronic hemodialysis patients. AB - BACKGROUND AND AIM: The aim of our study was to assess the coagulation factors as endothelial dysfunction markers and prospectively their association with thrombotic episodes in chronic hemodialysis patients. PATIENTS AND METHODS: Fifty four randomly selected patients on chronic hemodialysis (HD), 34 men and 20 women were included in this study. Their mean age was 56 years and the mean hemodialysis duration was 53.0861.92 months. The variations of tissue factor pathway inhibitor (TFPI), thrombomodulin (TM) and von Willebrand factor (vWF) were studied. The above-mentioned parameters were measured before and after HD session. Low molecular weight heparin (tinzaparin) was administered to all patients during hemodialysis. The results were compared with those obtained from 20 healthy volunteer-controls, age and sex matched. After the initial assessment, all patients were followed for a period of 15 months. RESULTS: Two patients experienced one hemorrhagic event each, regarding the upper and/or the lower gastrointestinal tract. Twenty patients showed at least one thrombotic episode. Eleven patients presented fistula thrombosis, four angina pectoris incidents and five thrombosis of the lower limbs. The rest of the patients did not experience any clinical symptomatology that could be attributed to coagulation disorders. Parameter differences between patients and controls were statistically significant (p<0.005 for TFPI and p<0.001 for TM and vWF) and were improved after hemodialysis session. The age and the elevated levels of the vWF were found to be significantly different (p<0.03 and p<0.02 respectively) between the groups of patients who experienced or not thrombotic episodes. CONCLUSIONS: Coagulation factors TFPI, TM, and vWF are increased in hemodialysis patients and the clinical disorders are mainly thrombotic episodes. The age of patients and the elevated levels of vWF are associated with the thrombotic incidents. Hemodialysis contributes in the improvement of these coagulation factors, which could be considered as biological risk markers of endothelial dysfunction in chronic HD patients. PMID- 20011090 TI - Acute respiratory admissions in Thessaloniki, Greece: 14-year follow-up. AB - BACKGROUND: Acute exacerbations of respiratory diseases are a common cause of hospitalization among infants and children. AIM: To assess the epidemiological characteristics of asthma, bronchiolitis and croup among hospitalized children in Thessaloniki, from 1990 to 2003 included. METHODS: Data from the patient registry with discharge diagnosis "asthma", "bronchiolitis" and "croup" were analyzed retrospectively, in five Paediatric Departments of Thessaloniki. Age and sex of the patients, as well as the month of admission were taken into consideration. RESULTS: A total of 8762 admissions of children (aged 3 months-14 years) with the diagnosis of asthma, "bronchiolitis" and "croup", were identified. Sex distribution was 65.86% males (64.86% bronchiolitis, 65.26% asthma and 70.31% croup). Asthma admissions decreased by 53.65%, croup admissions decreased by 4.73%, while bronchiolitis admissions increased by 25.03%, during the study period. A clear seasonal variation was found in all the three diseases, with the lowest incidence during summer months. Moreover there were two peaks for asthma (one during spring and a second during autumn), one peak for bronchiolitis (during winter early spring) and one peak for croup (during autumn). CONCLUSIONS: Paediatric asthma and croup admissions have declined during the last 14 years, in contrast with bronchiolitis admissions, which showed an increased tendency. More frequent use of inhaled steroids and induction of asthma education programs may have contributed to decreasing asthma admission rates. PMID- 20011091 TI - Guidelines for diagnosing and treating sleep related breathing disorders in adults and children (Part 2: treatment). PMID- 20011092 TI - Childhood obesity in relation to parental weight status in Greece. PMID- 20011093 TI - H1N1 influenza A infection: an update. PMID- 20011094 TI - Role of vitamin d in insulin secretion and insulin sensitivity for glucose homeostasis. AB - Vitamin D functions are not limited to skeletal health benefits and may extend to preservation of insulin secretion and insulin sensitivity. This review summarizes the literature related to potential vitamin D influences on glucose homeostasis and insulin sensitivity. Cross-sectional data provide some evidence that circulating 25-hydroxyvitamin D (25(OH)D) is inversely associated with insulin resistance, although direct measurements of insulin sensitivity are required for confirmation. Reported associations with insulin secretion, however, are contradictory. Available prospective studies support a protective influence of high 25(OH)D concentrations on type 2 diabetes mellitus risk. There is a general lack of consistency in vitamin D intervention outcomes on insulin secretion and sensitivity, likely due to differences in subject populations, length of interventions, and forms of vitamin D supplementation. Vitamin D receptor gene polymorphisms and vitamin D interactions with the insulin like growth factor system may further influence glucose homeostasis. The ambiguity of optimal vitamin D dosing regimens and optimal therapeutic concentrations of serum 25(OH)D limit available intervention studies. Future studies, including cross-sectional and prospective, should be performed in populations at high risk for both vitamin D deficiency and type 2 diabetes mellitus. Well-designed, placebo-controlled, randomized intervention studies are required to establish a true protective influence of vitamin D on glucose homeostasis. PMID- 20011095 TI - Possible Health Implications and Low Vitamin D Status during Childhood and Adolescence: An Updated Mini Review. AB - Vitamin D deficiency is common in the developing countries and exists in both childhood and adult life. The great importance of Vitamin D is the moderation of calcium (Ca) and phosphorus (P) homeostasis as well as the absorption of Ca. While insufficiency of vitamin D is a significant contributing factor to risk of rickets in childhood, it is possible that a more marginal deficiency of vitamin D during life span contribute to osteoporosis as well as potentially to the development and various other chronic diseases such as cardiovascular disease, cancer and diabetes. This paper reviews the metabolism, epidemiology, and treatment of vitamin D and calcium insufficiency as well as its relation to various diseases during childhood and adolescence. PMID- 20011096 TI - Vitamin d deficiency-a clinical spectrum: is there a symptomatic nonosteomalacic state? AB - Vitamin D deficiency is not uncommon even in the sunny land of India. Lack of sun exposure and inadequate oral intake are both responsible for vitamin D deficiency. This article provides a retrospective, examining the effects of Vitamin D deficiency in 71 patients. The study's inclusion criterion was low vitamin D level combined with musculoskeletal symptoms but without the presence of osteomalacia. All patients in this study were suspected to have vitamin D deficiency. The data were retrieved from the case-charts of patients seen between 1996 and 2001 at the rheumatology services of Hinduja Hospital, Mumbai, India. This study found no correlation between Vitamin D levels and symptoms, or between the severity of Vitamin D deficiency and the number of symptoms displayed. Subclinical vitamin D deficiency or preosteomalacic state was the term coined for individuals with vitamin D deficiency producing nonspecific musculoskeletal symptoms in the absence of clinical osteomalacia. PMID- 20011097 TI - Brigham fracture intervention team initiatives for hospital patients with hip fractures: a paradigm shift. AB - We designed, implemented, and revised the Brigham Fracture Intervention Team (B FIT) initiatives to improve in-hospital care of fracture (Fx) patients. Effectiveness was evaluated for 181 medical records of 4 cohorts in four successive years of consecutive patients who were admitted with a fragility hip Fx. The Discharge Initiative (DI) (computer-based) includes 1200 mg calcium and 1000 IU vitamin D(3) daily. The Admission Initiative (AI) was introduced one year later with reminders for serum 25OHD measurement, initiation of daily calcium (1200 mg) and vitamin D (800 IU), and an order for Endocrinology consultation, with an amendment for a computer-assisted reminder and a dose of D(2) (50 000 IU). Initially, the computer-based DI was more effective (67%) than the surgeon driven AI (33%, P < .001). After introduction of a computer-assisted reminder, AI effectiveness increased to 68%. The marked prevalence of vitamin D insufficiency reaffirms the importance of incorporating vitamin D recommendations in Fx care pathways. PMID- 20011098 TI - Concise syntheses of enantiomerically pure protected 4-hydroxypyroglutamic acid and 4-hydroxyproline from a nitroso-cyclopentadiene cycloadduct. AB - O-TBS-protected methyl trans-4-hydroxypyroglutamate and methyl trans-4 hydroxyproline ester were synthesized from nitroso-cyclopentadiene Diels-Alder cycloadducts. Enzymatic resolution of the key intermediate, 4-amino-cyclopent-2 enol, provides access to both L- and D- amino acids. PMID- 20011099 TI - A mouse model of the human Fragile X syndrome I304N mutation. AB - The mental retardation, autistic features, and behavioral abnormalities characteristic of the Fragile X mental retardation syndrome result from the loss of function of the RNA-binding protein FMRP. The disease is usually caused by a triplet repeat expansion in the 5'UTR of the FMR1 gene. This leads to loss of function through transcriptional gene silencing, pointing to a key function for FMRP, but precluding genetic identification of critical activities within the protein. Moreover, antisense transcripts (FMR4, ASFMR1) in the same locus have been reported to be silenced by the repeat expansion. Missense mutations offer one means of confirming a central role for FMRP in the disease, but to date, only a single such patient has been described. This patient harbors an isoleucine to asparagine mutation (I304N) in the second FMRP KH-type RNA-binding domain, however, this single case report was complicated because the patient harbored a superimposed familial liver disease. To address these issues, we have generated a new Fragile X Syndrome mouse model in which the endogenous Fmr1 gene harbors the I304N mutation. These mice phenocopy the symptoms of Fragile X Syndrome in the existing Fmr1-null mouse, as assessed by testicular size, behavioral phenotyping, and electrophysiological assays of synaptic plasticity. I304N FMRP retains some functions, but has specifically lost RNA binding and polyribosome association; moreover, levels of the mutant protein are markedly reduced in the brain specifically at a time when synapses are forming postnatally. These data suggest that loss of FMRP function, particularly in KH2-mediated RNA binding and in synaptic plasticity, play critical roles in pathogenesis of the Fragile X Syndrome and establish a new model for studying the disorder. PMID- 20011100 TI - SOS response induces persistence to fluoroquinolones in Escherichia coli. AB - Bacteria can survive antibiotic treatment without acquiring heritable antibiotic resistance. We investigated persistence to the fluoroquinolone ciprofloxacin in Escherichia coli. Our data show that a majority of persisters to ciprofloxacin were formed upon exposure to the antibiotic, in a manner dependent on the SOS gene network. These findings reveal an active and inducible mechanism of persister formation mediated by the SOS response, challenging the prevailing view that persisters are pre-existing and formed purely by stochastic means. SOS induced persistence is a novel mechanism by which cells can counteract DNA damage and promote survival to fluoroquinolones. This unique survival mechanism may be an important factor influencing the outcome of antibiotic therapy in vivo. PMID- 20011101 TI - Regulators of AWC-mediated olfactory plasticity in Caenorhabditis elegans. AB - While most sensory neurons will adapt to prolonged stimulation by down-regulating their responsiveness to the signal, it is not clear which events initiate long lasting sensory adaptation. Likewise, we are just beginning to understand how the physiology of the adapted cell is altered. Caenorhabditis elegans is inherently attracted to specific odors that are sensed by the paired AWC olfactory sensory neurons. The attraction diminishes if the animal experiences these odors for a prolonged period of time in the absence of food. The AWC neuron responds acutely to odor-exposure by closing calcium channels. While odortaxis requires a Galpha subunit protein, cGMP-gated channels, and guanylyl cyclases, adaptation to prolonged odor exposure requires nuclear entry of the cGMP-dependent protein kinase, EGL-4. We asked which candidate members of the olfactory signal transduction pathway promote nuclear entry of EGL-4 and which molecules might induce long-term adaptation downstream of EGL-4 nuclear entry. We found that initiation of long-term adaptation, as assessed by nuclear entry of EGL-4, is dependent on G-protein mediated signaling but is independent of fluxes in calcium levels. We show that long-term adaptation requires polyunsaturated fatty acids (PUFAs) that may act on the transient receptor potential (TRP) channel type V OSM 9 downstream of EGL-4 nuclear entry. We also present evidence that high diacylglycerol (DAG) levels block long-term adaptation without affecting EGL-4 nuclear entry. Our analysis provides a model for the process of long-term adaptation that occurs within the AWC neuron of C. elegans: G-protein signaling initiates long-lasting olfactory adaptation by promoting the nuclear entry of EGL 4, and once EGL-4 has entered the nucleus, processes such as PUFA activation of the TRP channel OSM-9 may dampen the output of the AWC neuron. PMID- 20011102 TI - Fine-scale variation and genetic determinants of alternative splicing across individuals. AB - Recently, thanks to the increasing throughput of new technologies, we have begun to explore the full extent of alternative pre-mRNA splicing (AS) in the human transcriptome. This is unveiling a vast layer of complexity in isoform-level expression differences between individuals. We used previously published splicing sensitive microarray data from lymphoblastoid cell lines to conduct an in-depth analysis on splicing efficiency of known and predicted exons. By combining publicly available AS annotation with a novel algorithm designed to search for AS, we show that many real AS events can be detected within the usually unexploited, speculative majority of the array and at significance levels much below standard multiple-testing thresholds, demonstrating that the extent of cis regulated differential splicing between individuals is potentially far greater than previously reported. Specifically, many genes show subtle but significant genetically controlled differences in splice-site usage. PCR validation shows that 42 out of 58 (72%) candidate gene regions undergo detectable AS, amounting to the largest scale validation of isoform eQTLs to date. Targeted sequencing revealed a likely causative SNP in most validated cases. In all 17 incidences where a SNP affected a splice-site region, in silico splice-site strength modeling correctly predicted the direction of the micro-array and PCR results. In 13 other cases, we identified likely causative SNPs disrupting predicted splicing enhancers. Using Fst and REHH analysis, we uncovered significant evidence that 2 putative causative SNPs have undergone recent positive selection. We verified the effect of five SNPs using in vivo minigene assays. This study shows that splicing differences between individuals, including quantitative differences in isoform ratios, are frequent in human populations and that causative SNPs can be identified using in silico predictions. Several cases affected disease-relevant genes and it is likely some of these differences are involved in phenotypic diversity and susceptibility to complex diseases. PMID- 20011103 TI - The GAAS metagenomic tool and its estimations of viral and microbial average genome size in four major biomes. AB - Metagenomic studies characterize both the composition and diversity of uncultured viral and microbial communities. BLAST-based comparisons have typically been used for such analyses; however, sampling biases, high percentages of unknown sequences, and the use of arbitrary thresholds to find significant similarities can decrease the accuracy and validity of estimates. Here, we present Genome relative Abundance and Average Size (GAAS), a complete software package that provides improved estimates of community composition and average genome length for metagenomes in both textual and graphical formats. GAAS implements a novel methodology to control for sampling bias via length normalization, to adjust for multiple BLAST similarities by similarity weighting, and to select significant similarities using relative alignment lengths. In benchmark tests, the GAAS method was robust to both high percentages of unknown sequences and to variations in metagenomic sequence read lengths. Re-analysis of the Sargasso Sea virome using GAAS indicated that standard methodologies for metagenomic analysis may dramatically underestimate the abundance and importance of organisms with small genomes in environmental systems. Using GAAS, we conducted a meta-analysis of microbial and viral average genome lengths in over 150 metagenomes from four biomes to determine whether genome lengths vary consistently between and within biomes, and between microbial and viral communities from the same environment. Significant differences between biomes and within aquatic sub-biomes (oceans, hypersaline systems, freshwater, and microbialites) suggested that average genome length is a fundamental property of environments driven by factors at the sub biome level. The behavior of paired viral and microbial metagenomes from the same environment indicated that microbial and viral average genome sizes are independent of each other, but indicative of community responses to stressors and environmental conditions. PMID- 20011104 TI - A genome-wide association study reveals variants in ARL15 that influence adiponectin levels. AB - The adipocyte-derived protein adiponectin is highly heritable and inversely associated with risk of type 2 diabetes mellitus (T2D) and coronary heart disease (CHD). We meta-analyzed 3 genome-wide association studies for circulating adiponectin levels (n = 8,531) and sought validation of the lead single nucleotide polymorphisms (SNPs) in 5 additional cohorts (n = 6,202). Five SNPs were genome-wide significant in their relationship with adiponectin (P< or =5x10( 8)). We then tested whether these 5 SNPs were associated with risk of T2D and CHD using a Bonferroni-corrected threshold of P< or =0.011 to declare statistical significance for these disease associations. SNPs at the adiponectin-encoding ADIPOQ locus demonstrated the strongest associations with adiponectin levels (P combined = 9.2x10(-19) for lead SNP, rs266717, n = 14,733). A novel variant in the ARL15 (ADP-ribosylation factor-like 15) gene was associated with lower circulating levels of adiponectin (rs4311394-G, P-combined = 2.9x10(-8), n = 14,733). This same risk allele at ARL15 was also associated with a higher risk of CHD (odds ratio [OR] = 1.12, P = 8.5x10(-6), n = 22,421) more nominally, an increased risk of T2D (OR = 1.11, P = 3.2x10(-3), n = 10,128), and several metabolic traits. Expression studies in humans indicated that ARL15 is well expressed in skeletal muscle. These findings identify a novel protein, ARL15, which influences circulating adiponectin levels and may impact upon CHD risk. PMID- 20011105 TI - Trade-off between positive and negative design of protein stability: from lattice models to real proteins. AB - Two different strategies for stabilizing proteins are (i) positive design in which the native state is stabilized and (ii) negative design in which competing non-native conformations are destabilized. Here, the circumstances under which one strategy might be favored over the other are explored in the case of lattice models of proteins and then generalized and discussed with regard to real proteins. The balance between positive and negative design of proteins is found to be determined by their average "contact-frequency", a property that corresponds to the fraction of states in the conformational ensemble of the sequence in which a pair of residues is in contact. Lattice model proteins with a high average contact-frequency are found to use negative design more than model proteins with a low average contact-frequency. A mathematical derivation of this result indicates that it is general and likely to hold also for real proteins. Comparison of the results of correlated mutation analysis for real proteins with typical contact-frequencies to those of proteins likely to have high contact frequencies (such as disordered proteins and proteins that are dependent on chaperonins for their folding) indicates that the latter tend to have stronger interactions between residues that are not in contact in their native conformation. Hence, our work indicates that negative design is employed when insufficient stabilization is achieved via positive design owing to high contact frequencies. PMID- 20011106 TI - An abundance of ubiquitously expressed genes revealed by tissue transcriptome sequence data. AB - The parts of the genome transcribed by a cell or tissue reflect the biological processes and functions it carries out. We characterized the features of mammalian tissue transcriptomes at the gene level through analysis of RNA deep sequencing (RNA-Seq) data across human and mouse tissues and cell lines. We observed that roughly 8,000 protein-coding genes were ubiquitously expressed, contributing to around 75% of all mRNAs by message copy number in most tissues. These mRNAs encoded proteins that were often intracellular, and tended to be involved in metabolism, transcription, RNA processing or translation. In contrast, genes for secreted or plasma membrane proteins were generally expressed in only a subset of tissues. The distribution of expression levels was broad but fairly continuous: no support was found for the concept of distinct expression classes of genes. Expression estimates that included reads mapping to coding exons only correlated better with qRT-PCR data than estimates which also included 3' untranslated regions (UTRs). Muscle and liver had the least complex transcriptomes, in that they expressed predominantly ubiquitous genes and a large fraction of the transcripts came from a few highly expressed genes, whereas brain, kidney and testis expressed more complex transcriptomes with the vast majority of genes expressed and relatively small contributions from the most expressed genes. mRNAs expressed in brain had unusually long 3'UTRs, and mean 3'UTR length was higher for genes involved in development, morphogenesis and signal transduction, suggesting added complexity of UTR-based regulation for these genes. Our results support a model in which variable exterior components feed into a large, densely connected core composed of ubiquitously expressed intracellular proteins. PMID- 20011107 TI - Challenges predicting ligand-receptor interactions of promiscuous proteins: the nuclear receptor PXR. AB - Transcriptional regulation of some genes involved in xenobiotic detoxification and apoptosis is performed via the human pregnane X receptor (PXR) which in turn is activated by structurally diverse agonists including steroid hormones. Activation of PXR has the potential to initiate adverse effects, altering drug pharmacokinetics or perturbing physiological processes. Reliable computational prediction of PXR agonists would be valuable for pharmaceutical and toxicological research. There has been limited success with structure-based modeling approaches to predict human PXR activators. Slightly better success has been achieved with ligand-based modeling methods including quantitative structure-activity relationship (QSAR) analysis, pharmacophore modeling and machine learning. In this study, we present a comprehensive analysis focused on prediction of 115 steroids for ligand binding activity towards human PXR. Six crystal structures were used as templates for docking and ligand-based modeling approaches (two-, three-, four- and five-dimensional analyses). The best success at external prediction was achieved with 5D-QSAR. Bayesian models with FCFP_6 descriptors were validated after leaving a large percentage of the dataset out and using an external test set. Docking of ligands to the PXR structure co-crystallized with hyperforin had the best statistics for this method. Sulfated steroids (which are activators) were consistently predicted as non-activators while, poorly predicted steroids were docked in a reverse mode compared to 5alpha-androstan-3beta-ol. Modeling of human PXR represents a complex challenge by virtue of the large, flexible ligand-binding cavity. This study emphasizes this aspect, illustrating modest success using the largest quantitative data set to date and multiple modeling approaches. PMID- 20011108 TI - ON/OFF and beyond--a boolean model of apoptosis. AB - Apoptosis is regulated by several signaling pathways which are extensively linked by crosstalks. Boolean or logical modeling has become a promising approach to capture the qualitative behavior of such complex networks. Here we built a large scale literature-based Boolean model of the central intrinsic and extrinsic apoptosis pathways as well as pathways connected with them. The model responds to several external stimuli such as Fas ligand, TNF-alpha, UV-B irradiation, interleukin-1beta and insulin. Timescales and multi-value node logic were used and turned out to be indispensable to reproduce the behavior of the apoptotic network. The coherence of the model was experimentally validated. Thereby an UV-B dose-effect is shown for the first time in mouse hepatocytes. Analysis of the model revealed a tight regulation emerging from high connectivity and spanning crosstalks and a particular importance of feedback loops. An unexpected feedback from Smac release to RIP could further increase complex II formation. The introduced Boolean model provides a comprehensive and coherent description of the apoptosis network behavior. It gives new insights into the complex interplay of pro- and antiapoptotic factors and can be easily expanded to other signaling pathways. PMID- 20011109 TI - Annotation error in public databases: misannotation of molecular function in enzyme superfamilies. AB - Due to the rapid release of new data from genome sequencing projects, the majority of protein sequences in public databases have not been experimentally characterized; rather, sequences are annotated using computational analysis. The level of misannotation and the types of misannotation in large public databases are currently unknown and have not been analyzed in depth. We have investigated the misannotation levels for molecular function in four public protein sequence databases (UniProtKB/Swiss-Prot, GenBank NR, UniProtKB/TrEMBL, and KEGG) for a model set of 37 enzyme families for which extensive experimental information is available. The manually curated database Swiss-Prot shows the lowest annotation error levels (close to 0% for most families); the two other protein sequence databases (GenBank NR and TrEMBL) and the protein sequences in the KEGG pathways database exhibit similar and surprisingly high levels of misannotation that average 5%-63% across the six superfamilies studied. For 10 of the 37 families examined, the level of misannotation in one or more of these databases is >80%. Examination of the NR database over time shows that misannotation has increased from 1993 to 2005. The types of misannotation that were found fall into several categories, most associated with "overprediction" of molecular function. These results suggest that misannotation in enzyme superfamilies containing multiple families that catalyze different reactions is a larger problem than has been recognized. Strategies are suggested for addressing some of the systematic problems contributing to these high levels of misannotation. PMID- 20011110 TI - Early evolution of conserved regulatory sequences associated with development in vertebrates. AB - Comparisons between diverse vertebrate genomes have uncovered thousands of highly conserved non-coding sequences, an increasing number of which have been shown to function as enhancers during early development. Despite their extreme conservation over 500 million years from humans to cartilaginous fish, these elements appear to be largely absent in invertebrates, and, to date, there has been little understanding of their mode of action or the evolutionary processes that have modelled them. We have now exploited emerging genomic sequence data for the sea lamprey, Petromyzon marinus, to explore the depth of conservation of this type of element in the earliest diverging extant vertebrate lineage, the jawless fish (agnathans). We searched for conserved non-coding elements (CNEs) at 13 human gene loci and identified lamprey elements associated with all but two of these gene regions. Although markedly shorter and less well conserved than within jawed vertebrates, identified lamprey CNEs are able to drive specific patterns of expression in zebrafish embryos, which are almost identical to those driven by the equivalent human elements. These CNEs are therefore a unique and defining characteristic of all vertebrates. Furthermore, alignment of lamprey and other vertebrate CNEs should permit the identification of persistent sequence signatures that are responsible for common patterns of expression and contribute to the elucidation of the regulatory language in CNEs. Identifying the core regulatory code for development, common to all vertebrates, provides a foundation upon which regulatory networks can be constructed and might also illuminate how large conserved regulatory sequence blocks evolve and become fixed in genomic DNA. PMID- 20011111 TI - What do transitive inference and class inclusion have in common? Categorical (co)products and cognitive development. AB - Transitive inference, class inclusion and a variety of other inferential abilities have strikingly similar developmental profiles-all are acquired around the age of five. Yet, little is known about the reasons for this correspondence. Category theory was invented as a formal means of establishing commonalities between various mathematical structures. We use category theory to show that transitive inference and class inclusion involve dual mathematical structures, called product and coproduct. Other inferential tasks with similar developmental profiles, including matrix completion, cardinality, dimensional changed card sorting, balance-scale (weight-distance integration), and Theory of Mind also involve these structures. By contrast, (co)products are not involved in the behaviours exhibited by younger children on these tasks, or simplified versions that are within their ability. These results point to a fundamental cognitive principle under development during childhood that is the capacity to compute (co)products in the categorical sense. PMID- 20011112 TI - The synaptonemal complex protein Zip1 promotes bi-orientation of centromeres at meiosis I. AB - In meiosis I, homologous chromosomes become paired and then separate from one another to opposite poles of the spindle. In humans, errors in this process are a leading cause of birth defects, mental retardation, and infertility. In most organisms, crossing-over, or exchange, between the homologous partners provides a link that promotes their proper, bipolar, attachment to the spindle. Attachment of both partners to the same pole can sometimes be corrected during a delay that is triggered by the spindle checkpoint. Studies of non-exchange chromosomes have shown that centromere pairing serves as an alternative to exchange by orienting the centromeres for proper microtubule attachment. Here, we demonstrate a new role for the synaptonemal complex protein Zip1. Zip1 localizes to the centromeres of non-exchange chromosomes in pachytene and mediates centromere pairing and segregation of the partners at meiosis I. Exchange chromosomes were also found to experience Zip1-dependent pairing at their centromeres. Zip1 was found to persist at centromeres, after synaptonemal complex disassembly, remaining there until microtubule attachment. Disruption of this centromere pairing, in spindle checkpoint mutants, randomized the segregation of exchange chromosomes. These results demonstrate that Zip1-mediated pairing of exchange chromosome centromeres promotes an initial, bipolar attachment of microtubules. This activity of Zip1 lessens the load on the spindle checkpoint, greatly reducing the chance that the cell will exit the checkpoint delay with an improperly oriented chromosome pair. Thus exchange, the spindle checkpoint, and centromere pairing are complementary mechanisms that ensure the proper segregation of homologous partners at meiosis I. PMID- 20011113 TI - Comprehensive functional analysis of Mycobacterium tuberculosis toxin-antitoxin systems: implications for pathogenesis, stress responses, and evolution. AB - Toxin-antitoxin (TA) systems, stress-responsive genetic elements ubiquitous in microbial genomes, are unusually abundant in the major human pathogen Mycobacterium tuberculosis. Why M. tuberculosis has so many TA systems and what role they play in the unique biology of the pathogen is unknown. To address these questions, we have taken a comprehensive approach to identify and functionally characterize all the TA systems encoded in the M. tuberculosis genome. Here we show that 88 putative TA system candidates are present in M. tuberculosis, considerably more than previously thought. Comparative genomic analysis revealed that the vast majority of these systems are conserved in the M. tuberculosis complex (MTBC), but largely absent from other mycobacteria, including close relatives of M. tuberculosis. We found that many of the M. tuberculosis TA systems are located within discernable genomic islands and were thus likely acquired recently via horizontal gene transfer. We discovered a novel TA system located in the core genome that is conserved across the genus, suggesting that it may fulfill a role common to all mycobacteria. By expressing each of the putative TA systems in M. smegmatis, we demonstrate that 30 encode a functional toxin and its cognate antitoxin. We show that the toxins of the largest family of TA systems, VapBC, act by inhibiting translation via mRNA cleavage. Expression profiling demonstrated that four systems are specifically activated during stresses likely encountered in vivo, including hypoxia and phagocytosis by macrophages. The expansion and maintenance of TA genes in the MTBC, coupled with the finding that a subset is transcriptionally activated by stress, suggests that TA systems are important for M. tuberculosis pathogenesis. PMID- 20011114 TI - Lymphangiogenesis and lymphatic remodeling induced by filarial parasites: implications for pathogenesis. AB - Even in the absence of an adaptive immune system in murine models, lymphatic dilatation and dysfunction occur in filarial infections, although severe irreversible lymphedema and elephantiasis appears to require an intact adaptive immune response in human infections. To address how filarial parasites and their antigens influence the lymphatics directly, human lymphatic endothelial cells were exposed to filarial antigens, live parasites, or infected patient serum. Live filarial parasites or filarial antigens induced both significant LEC proliferation and differentiation into tube-like structures in vitro. Moreover, serum from patently infected (microfilaria positive) patients and those with longstanding chronic lymphatic obstruction induced significantly increased LEC proliferation compared to sera from uninfected individuals. Differentiation of LEC into tube-like networks was found to be associated with significantly increased levels of matrix metalloproteases and inhibition of their TIMP inhibitors (Tissue inhibitors of matrix metalloproteases). Comparison of global gene expression induced by live parasites in LEC to parasite-unexposed LEC demonstrated that filarial parasites altered the expression of those genes involved in cellular organization and development as well as those associated with junction adherence pathways that in turn decreased trans-endothelial transport as assessed by FITC-Dextran. The data suggest that filarial parasites directly induce lymphangiogenesis and lymphatic differentiation and provide insight into the mechanisms underlying the pathology seen in lymphatic filariasis. PMID- 20011117 TI - The eureka moment: an interview with Sir Alec Jeffreys. Interview by Jane Gitschier. PMID- 20011116 TI - Population structure induces a symmetry breaking favoring the emergence of cooperation. AB - The evolution of cooperation described in terms of simple two-person interactions has received considerable attention in recent years, where several key results were obtained. Among those, it is now well established that the web of social interaction networks promotes the emergence of cooperation when modeled in terms of symmetric two-person games. Up until now, however, the impacts of the heterogeneity of social interactions into the emergence of cooperation have not been fully explored, as other aspects remain to be investigated. Here we carry out a study employing the simplest example of a prisoner's dilemma game in which the benefits collected by the participants may be proportional to the costs expended. We show that the heterogeneous nature of the social network naturally induces a symmetry breaking of the game, as contributions made by cooperators may become contingent on the social context in which the individual is embedded. A new, numerical, mean-field analysis reveals that prisoner's dilemmas on networks no longer constitute a defector dominance dilemma--instead, individuals engage effectively in a general coordination game. We find that the symmetry breaking induced by population structure profoundly affects the evolutionary dynamics of cooperation, dramatically enhancing the feasibility of cooperators: cooperation blooms when each cooperator contributes the same cost, equally shared among the plethora of games in which she participates. This work provides clear evidence that, while individual rational reasoning may hinder cooperative actions, the intricate nature of social interactions may effectively transform a local dilemma of cooperation into a global coordination problem. PMID- 20011115 TI - CD14 signaling restrains chronic inflammation through induction of p38-MAPK/SOCS dependent tolerance. AB - Current thinking emphasizes the primacy of CD14 in facilitating recognition of microbes by certain TLRs to initiate pro-inflammatory signaling events and the importance of p38-MAPK in augmenting such responses. Herein, this paradigm is challenged by demonstrating that recognition of live Borrelia burgdorferi not only triggers an inflammatory response in the absence of CD14, but one that is, in part, a consequence of altered PI3K/AKT/p38-MAPK signaling and impaired negative regulation of TLR2. CD14 deficiency results in increased localization of PI3K to lipid rafts, hyperphosphorylation of AKT, and reduced activation of p38. Such aberrant signaling leads to decreased negative regulation by SOCS1, SOCS3, and CIS, thereby compromising the induction of tolerance in macrophages and engendering more severe and persistent inflammatory responses to B. burgdorferi. Importantly, these altered signaling events and the higher cytokine production observed can be mimicked through shRNA and pharmacological inhibition of p38 activity in CD14-expressing macrophages. Perturbation of this CD14/p38-MAPK dependent immune regulation may underlie development of infectious chronic inflammatory syndromes. PMID- 20011118 TI - Discovery of candidate disease genes in ENU-induced mouse mutants by large-scale sequencing, including a splice-site mutation in nucleoredoxin. AB - An accurate and precisely annotated genome assembly is a fundamental requirement for functional genomic analysis. Here, the complete DNA sequence and gene annotation of mouse Chromosome 11 was used to test the efficacy of large-scale sequencing for mutation identification. We re-sequenced the 14,000 annotated exons and boundaries from over 900 genes in 41 recessive mutant mouse lines that were isolated in an N-ethyl-N-nitrosourea (ENU) mutation screen targeted to mouse Chromosome 11. Fifty-nine sequence variants were identified in 55 genes from 31 mutant lines. 39% of the lesions lie in coding sequences and create primarily missense mutations. The other 61% lie in noncoding regions, many of them in highly conserved sequences. A lesion in the perinatal lethal line l11Jus13 alters a consensus splice site of nucleoredoxin (Nxn), inserting 10 amino acids into the resulting protein. We conclude that point mutations can be accurately and sensitively recovered by large-scale sequencing, and that conserved noncoding regions should be included for disease mutation identification. Only seven of the candidate genes we report have been previously targeted by mutation in mice or rats, showing that despite ongoing efforts to functionally annotate genes in the mammalian genome, an enormous gap remains between phenotype and function. Our data show that the classical positional mapping approach of disease mutation identification can be extended to large target regions using high-throughput sequencing. PMID- 20011119 TI - Cortical gamma rhythms modulate NMDAR-mediated spike timing dependent plasticity in a biophysical model. AB - Spike timing dependent plasticity (STDP) has been observed experimentally in vitro and is a widely studied neural algorithm for synaptic modification. While the functional role of STDP has been investigated extensively, the effect of rhythms on the precise timing of STDP has not been characterized as well. We use a simplified biophysical model of a cortical network that generates pyramidal interneuronal gamma rhythms (PING). Plasticity via STDP is investigated at the excitatory pyramidal cell synapse from a gamma frequency (30-90 Hz) input independent of the network gamma rhythm. The input may represent a corticocortical or an information-specific thalamocortical connection. This synapse is mediated by N-methyl-D-aspartate receptor mediated (NMDAR) currents. For distinct network and input frequencies, the model shows robust frequency regimes of potentiation and depression, providing a mechanism by which responses to certain inputs can potentiate while responses to other inputs depress. For potentiating regimes, the model suggests an optimal amount and duration of plasticity that can occur, which depends on the time course for the decay of the postsynaptic NMDAR current. Prolonging the duration of the input beyond this optimal time results in depression. Inserting pauses in the input can increase the total potentiation. The optimal pause length corresponds to the decay time of the NMDAR current. Thus, STDP in this model provides a mechanism for potentiation and depression depending on input frequency and suggests that the slow NMDAR current decay helps to regulate the optimal amplitude and duration of the plasticity. The optimal pause length is comparable to the time scale of the negative phase of a modulatory theta rhythm, which may pause gamma rhythm spiking. Our pause results may suggest a novel role for this theta rhythm in plasticity. Finally, we discuss our results in the context of auditory thalamocortical plasticity. PMID- 20011121 TI - Retrotransposon silencing by piRNAs: ping-pong players mark their sub-cellular boundaries. PMID- 20011120 TI - Interaction of HP1 and Brg1/Brm with the globular domain of histone H3 is required for HP1-mediated repression. AB - The heterochromatin-enriched HP1 proteins play a critical role in regulation of transcription. These proteins contain two related domains known as the chromo- and the chromoshadow-domain. The chromo-domain binds histone H3 tails methylated on lysine 9. However, in vivo and in vitro experiments have shown that the affinity of HP1 proteins to native methylated chromatin is relatively poor and that the opening of chromatin occurring during DNA replication facilitates their binding to nucleosomes. These observations prompted us to investigate whether HP1 proteins have additional histone binding activities, envisioning also affinity for regions potentially occluded by the nucleosome structure. We find that the chromoshadow-domain interacts with histone H3 in a region located partially inside the nucleosomal barrel at the entry/exit point of the nucleosome. Interestingly, this region is also contacted by the catalytic subunits of the human SWI/SNF complex. In vitro, efficient SWI/SNF remodeling requires this contact and is inhibited in the presence of HP1 proteins. The antagonism between SWI/SNF and HP1 proteins is also observed in vivo on a series of interferon regulated genes. Finally, we show that SWI/SNF activity favors loading of HP1 proteins to chromatin both in vivo and in vitro. Altogether, our data suggest that HP1 chromoshadow-domains can benefit from the opening of nucleosomal structures to bind chromatin and that HP1 proteins use this property to detect and arrest unwanted chromatin remodeling. PMID- 20011122 TI - Network properties of robust immunity in plants. AB - Two modes of plant immunity against biotrophic pathogens, Effector Triggered Immunity (ETI) and Pattern-Triggered Immunity (PTI), are triggered by recognition of pathogen effectors and Microbe-Associated Molecular Patterns (MAMPs), respectively. Although the jasmonic acid (JA)/ethylene (ET) and salicylic acid (SA) signaling sectors are generally antagonistic and important for immunity against necrotrophic and biotrophic pathogens, respectively, their precise roles and interactions in ETI and PTI have not been clear. We constructed an Arabidopsis dde2/ein2/pad4/sid2-quadruple mutant. DDE2, EIN2, and SID2 are essential components of the JA, ET, and SA sectors, respectively. The pad4 mutation affects the SA sector and a poorly characterized sector. Although the ETI triggered by the bacterial effector AvrRpt2 (AvrRpt2-ETI) and the PTI triggered by the bacterial MAMP flg22 (flg22-PTI) were largely intact in plants with mutations in any one of these genes, they were mostly abolished in the quadruple mutant. For the purposes of this study, AvrRpt2-ETI and flg22-PTI were measured as relative growth of Pseudomonas syringae bacteria within leaves. Immunity to the necrotrophic fungal pathogen Alternaria brassicicola was also severely compromised in the quadruple mutant. Quantitative measurements of the immunity levels in all combinatorial mutants and wild type allowed us to estimate the effects of the wild-type genes and their interactions on the immunity by fitting a mixed general linear model. This signaling allocation analysis showed that, contrary to current ideas, each of the JA, ET, and SA signaling sectors can positively contribute to immunity against both biotrophic and necrotrophic pathogens. The analysis also revealed that while flg22-PTI and AvrRpt2-ETI use a highly overlapping signaling network, the way they use the common network is very different: synergistic relationships among the signaling sectors are evident in PTI, which may amplify the signal; compensatory relationships among the sectors dominate in ETI, explaining the robustness of ETI against genetic and pathogenic perturbations. PMID- 20011123 TI - Integrin clustering is driven by mechanical resistance from the glycocalyx and the substrate. AB - Integrins have emerged as key sensory molecules that translate chemical and physical cues from the extracellular matrix (ECM) into biochemical signals that regulate cell behavior. Integrins function by clustering into adhesion plaques, but the molecular mechanisms that drive integrin clustering in response to interaction with the ECM remain unclear. To explore how deformations in the cell ECM interface influence integrin clustering, we developed a spatial-temporal simulation that integrates the micro-mechanics of the cell, glycocalyx, and ECM with a simple chemical model of integrin activation and ligand interaction. Due to mechanical coupling, we find that integrin-ligand interactions are highly cooperative, and this cooperativity is sufficient to drive integrin clustering even in the absence of cytoskeletal crosslinking or homotypic integrin-integrin interactions. The glycocalyx largely mediates this cooperativity and hence may be a key regulator of integrin function. Remarkably, integrin clustering in the model is naturally responsive to the chemical and physical properties of the ECM, including ligand density, matrix rigidity, and the chemical affinity of ligand for receptor. Consistent with experimental observations, we find that integrin clustering is robust on rigid substrates with high ligand density, but is impaired on substrates that are highly compliant or have low ligand density. We thus demonstrate how integrins themselves could function as sensory molecules that begin sensing matrix properties even before large multi-molecular adhesion complexes are assembled. PMID- 20011124 TI - A multi-component model of the developing retinocollicular pathway incorporating axonal and synaptic growth. AB - During development, neurons extend axons to different brain areas and produce stereotypical patterns of connections. The mechanisms underlying this process have been intensively studied in the visual system, where retinal neurons form retinotopic maps in the thalamus and superior colliculus. The mechanisms active in map formation include molecular guidance cues, trophic factor release, spontaneous neural activity, spike-timing dependent plasticity (STDP), synapse creation and retraction, and axon growth, branching and retraction. To investigate how these mechanisms interact, a multi-component model of the developing retinocollicular pathway was produced based on phenomenological approximations of each of these mechanisms. Core assumptions of the model were that the probabilities of axonal branching and synaptic growth are highest where the combined influences of chemoaffinity and trophic factor cues are highest, and that activity-dependent release of trophic factors acts to stabilize synapses. Based on these behaviors, model axons produced morphologically realistic growth patterns and projected to retinotopically correct locations in the colliculus. Findings of the model include that STDP, gradient detection by axonal growth cones and lateral connectivity among collicular neurons were not necessary for refinement, and that the instructive cues for axonal growth appear to be mediated first by molecular guidance and then by neural activity. Although complex, the model appears to be insensitive to variations in how the component developmental mechanisms are implemented. Activity, molecular guidance and the growth and retraction of axons and synapses are common features of neural development, and the findings of this study may have relevance beyond organization in the retinocollicular pathway. PMID- 20011125 TI - The T3SS effector EspT defines a new category of invasive enteropathogenic E. coli (EPEC) which form intracellular actin pedestals. AB - Enteropathogenic Escherichia coli (EPEC) strains are defined as extracellular pathogens which nucleate actin rich pedestal-like membrane extensions on intestinal enterocytes to which they intimately adhere. EPEC infection is mediated by type III secretion system effectors, which modulate host cell signaling. Recently we have shown that the WxxxE effector EspT activates Rac1 and Cdc42 leading to formation of membrane ruffles and lamellipodia. Here we report that EspT-induced membrane ruffles facilitate EPEC invasion into non-phagocytic cells in a process involving Rac1 and Wave2. Internalized EPEC resides within a vacuole and Tir is localized to the vacuolar membrane, resulting in actin polymerization and formation of intracellular pedestals. To the best of our knowledge this is the first time a pathogen has been shown to induce formation of actin comets across a vacuole membrane. Moreover, our data breaks the dogma of EPEC as an extracellular pathogen and defines a new category of invasive EPEC. PMID- 20011126 TI - Degradation of host sphingomyelin is essential for Leishmania virulence. AB - In eukaryotes, sphingolipids (SLs) are important membrane components and powerful signaling molecules. In Leishmania, the major group of SLs is inositol phosphorylceramide (IPC), which is common in yeast and Trypanosomatids but absent in mammals. In contrast, sphingomyelin is not synthesized by Leishmania but is abundant in mammals. In the promastigote stage in vitro, Leishmania use SL metabolism as a major pathway to produce ethanolamine (EtN), a metabolite essential for survival and differentiation from non-virulent procyclics to highly virulent metacyclics. To further probe SL metabolism, we identified a gene encoding a putative neutral sphingomyelinase (SMase) and/or IPC hydrolase (IPCase), designated ISCL (Inositol phosphoSphingolipid phospholipase C-Like). Despite the lack of sphingomyelin synthesis, L. major promastigotes exhibited a potent SMase activity which was abolished upon deletion of ISCL, and increased following over-expression by episomal complementation. ISCL-dependent activity with sphingomyelin was about 20 fold greater than that seen with IPC. Null mutants of ISCL (iscl(-)) showed modest accumulation of IPC, but grew and differentiated normally in vitro. Interestingly, iscl(-) mutants did not induce lesion pathology in the susceptible BALB/c mice, yet persisted indefinitely at low levels at the site of infection. Notably, the acute virulence of iscl(-) was completely restored by the expression of ISCL or heterologous mammalian or fungal SMases, but not by fungal proteins exhibiting only IPCase activity. Together, these findings strongly suggest that degradation of host-derived sphingomyelin plays a pivotal role in the proliferation of Leishmania in mammalian hosts and the manifestation of acute disease pathology. PMID- 20011128 TI - Recent Progress in Syntheses and Applications of Dumbbell-like Nanoparticles. AB - This paper reviews the recent research progress in syntheses and applications of dumbbell-like nanoparticles. It first describes the general synthesis of dumbbell like nanoparticles containing noble metal and magnetic NPs/or quantum dots. It then outlines the interesting optical and magnetic properties found in these dumbbell nanoparticles. The review further highlights several exciting application potentials of these nanoparticles in catalysis and biomedicine. PMID- 20011127 TI - Functional memory B cells and long-lived plasma cells are generated after a single Plasmodium chabaudi infection in mice. AB - Antibodies have long been shown to play a critical role in naturally acquired immunity to malaria, but it has been suggested that Plasmodium-specific antibodies in humans may not be long lived. The cellular mechanisms underlying B cell and antibody responses are difficult to study in human infections; therefore, we have investigated the kinetics, duration and characteristics of the Plasmodium-specific memory B cell response in an infection of P. chabaudi in mice. Memory B cells and plasma cells specific for the C-terminal region of Merozoite Surface Protein 1 were detectable for more than eight months following primary infection. Furthermore, a classical memory response comprised predominantly of the T-cell dependent isotypes IgG2c, IgG2b and IgG1 was elicited upon rechallenge with the homologous parasite, confirming the generation of functional memory B cells. Using cyclophosphamide treatment to discriminate between long-lived and short-lived plasma cells, we demonstrated long-lived cells secreting Plasmodium-specific IgG in both bone marrow and in spleens of infected mice. The presence of these long-lived cells was independent of the presence of chronic infection, as removal of parasites with anti-malarial drugs had no impact on their numbers. Thus, in this model of malaria, both functional Plasmodium specific memory B cells and long-lived plasma cells can be generated, suggesting that defects in generating these cell populations may not be the reason for generating short-lived antibody responses. PMID- 20011130 TI - Investigating the Roles of Knowledge and Cognitive Abilities in Older Adult Information Seeking on the Web. AB - This study investigated the influences of knowledge, particularly Internet, Web browser, and search engine knowledge, as well as cognitive abilities on older adult information seeking on the Internet. The emphasis on aspects of cognition was informed by a modeling framework of search engine information-seeking behavior. Participants from two older age groups were recruited: twenty people in a younger-old group (ages 60-70) and twenty people in an older-old group (ages 71 85). Ten younger adults (ages 18-39) served as a comparison group. All participants had at least some Internet search experience. The experimental task consisted of six realistic search problems, all involving information related to health and well-being and which varied in degree of complexity. The results indicated that though necessary, Internet-related knowledge was not sufficient in explaining information-seeking performance, and suggested that a combination of both knowledge and key cognitive abilities is important for successful information seeking. In addition, the cognitive abilities that were found to be critical for task performance depended on the search problem's complexity. Also, significant differences in task performance between the younger and the two older age groups were found on complex, but not on simple problems. Overall, the results from this study have implications for instructing older adults on Internet information seeking and for the design of Web sites. PMID- 20011129 TI - Autonomic function in hypertension; role of genetic variation at the catecholamine storage vesicle protein chromogranin B. PMID- 20011131 TI - Depression in Age-Related Macular Degeneration. AB - Age-related macular degeneration (AMD) is a major cause of disability in the elderly, substantially degrades the quality of their lives, and is a risk factor for depression. Rates of depression in AMD are substantially greater than those found in the general population of older people, and are on par with those of other chronic and disabling diseases. This article discusses the effect of depression on vision-related disability in patients with AMD, suggests methods for screening for depression, and summarizes interventions for preventing depression in this high-risk group. PMID- 20011132 TI - A CONSTRAINED NON-RIGID REGISTRATION ALGORITHM FOR APPLICATION IN PROSTATE RADIOTHERAPY. AB - This paper presents a novel free-form deformation registration algorithm with non rigid constraints to capture the transformation between the planning day and treatment day CT images used for external beam radiotherapy for prostate cancer. The algorithm is constrained to the predetermined motion of a segmented organ, which is described by an injective free-form deformation (FFD) based on B splines. The end goal is for the injective transformation to be used to update the radiotherapy plan to take into account bone and soft tissue deformation. The results of the algorithm have been compared to those achieved using rigid and fully non-rigid registration. The results clearly indicate that the constrained non-rigid registration algorithm presented in this paper performed much better at capturing the motion of the constrained organ, the bladder in this case, than the rigid or fully non-rigid registration algorithms. PMID- 20011133 TI - Keeping families engaged: the effects of home-based family therapy enhanced with experiential activities. AB - Engagement of high-risk adolescents and their families in treatment is a considerable challenge for service providers and agencies. Despite its importance, little research has been conducted that explores this important treatment process. To address this gap, a test of an innovative method to improve engagement in family therapy was undertaken. Findings of this study of 42 intervention group families and 41 comparison group families (N=83) suggest that augmenting in-home family therapy with short and creative experiential activities can significantly increase engagement and retention in treatment. Further research of engagement as a mechanism of change in family-based treatment is needed. PMID- 20011134 TI - Cloning, expression and purification of an acetoacetyl CoA thiolase from sunflower cotyledon. AB - Thiolase I and II coexist as part of the glyoxysomal beta-oxidation system in sunflower (Helianthus annuus L.) cotyledons, the only system shown to have both forms. The importance of thiolases can be underscored not only by their ubiquity, but also by their involvement in a wide variety of processes in plants, animals and bacteria. Here we describe the cloning, expression and purification of acetoacetyl CoA thiolase (AACT) in enzymatically active form. Use of the extensive amount of sequence information from the databases facilitated the efficient generation of the gene-specific primers used in the RACE protocols. The recombinant AACT (1233 bp) shares 75% similarity with other plant AACTs. Comparison of specific activity of this recombinant AACT to a previously reported enzyme purified from primary sunflower cotyledon tissue was very similar (263 nkat/mg protein vs 220 nkat/mg protein, respectively). Combining the most pure fractions from the affinity column, the enzyme was purified 88-fold with a 55% yield of the enzymatically active, 47 kDa AACT. PMID- 20011136 TI - A comparison of the effects of three GM corn varieties on mammalian health. AB - We present for the first time a comparative analysis of blood and organ system data from trials with rats fed three main commercialized genetically modified (GM) maize (NK 603, MON 810, MON 863), which are present in food and feed in the world. NK 603 has been modified to be tolerant to the broad spectrum herbicide Roundup and thus contains residues of this formulation. MON 810 and MON 863 are engineered to synthesize two different Bt toxins used as insecticides. Approximately 60 different biochemical parameters were classified per organ and measured in serum and urine after 5 and 14 weeks of feeding. GM maize-fed rats were compared first to their respective isogenic or parental non-GM equivalent control groups. This was followed by comparison to six reference groups, which had consumed various other non-GM maize varieties. We applied nonparametric methods, including multiple pairwise comparisons with a False Discovery Rate approach. Principal Component Analysis allowed the investigation of scattering of different factors (sex, weeks of feeding, diet, dose and group). Our analysis clearly reveals for the 3 GMOs new side effects linked with GM maize consumption, which were sex- and often dose-dependent. Effects were mostly associated with the kidney and liver, the dietary detoxifying organs, although different between the 3 GMOs. Other effects were also noticed in the heart, adrenal glands, spleen and haematopoietic system. We conclude that these data highlight signs of hepatorenal toxicity, possibly due to the new pesticides specific to each GM corn. In addition, unintended direct or indirect metabolic consequences of the genetic modification cannot be excluded. PMID- 20011135 TI - A receptor and binding protein interplay in the detection of a distinct pheromone component in the silkmoth Antheraea polyphemus. AB - Male moths respond to conspecific female-released pheromones with remarkable sensitivity and specificity, due to highly specialized chemosensory neurons in their antennae. In Antheraea silkmoths, three types of sensory neurons have been described, each responsive to one of three pheromone components. Since also three different pheromone binding proteins (PBPs) have been identified, the antenna of Antheraea seems to provide a unique model system for detailed analyzes of the interplay between the various elements underlying pheromone reception. Efforts to identify pheromone receptors of Antheraea polyphemus have led to the identification of a candidate pheromone receptor (ApolOR1). This receptor was found predominantly expressed in male antennae, specifically in neurons located beneath pheromone-sensitive sensilla trichodea. The ApolOR1-expressing cells were found to be surrounded by supporting cells co-expressing all three ApolPBPs. The response spectrum of ApolOR1 was assessed by means of calcium imaging using HEK293-cells stably expressing the receptor. It was found that at nanomolar concentrations ApolOR1-cells responded to all three pheromones when the compounds were solubilized by DMSO and also when DMSO was substituted by one of the three PBPs. However, at picomolar concentrations, cells responded only in the presence of the subtype ApolPBP2 and the pheromone (E,Z)-6,11-hexadecadienal. These results are indicative of a specific interplay of a distinct pheromone component with an appropriate binding protein and its related receptor subtype, which may be considered as basis for the remarkable sensitivity and specificity of the pheromone detection system. PMID- 20011137 TI - Physiological and oncogenic Aurora-A pathway. AB - Aurora family of protein kinases have emerged as crucial factors of, not only mitosis and cytokinesis, but also human carcinogenesis. Among these family members is Aurora-A that is frequently overexpressed in varieties of human cancer. Both in vitro and in vivo studies demonstrated that Aurora-A induces tumorigenesis through genome instability. These studies have further shown that cell signaling cross-talk between Aurora-A and other cellular proteins are essential for fully-transformed phenotypes. This review summarizes recent progress of Aurora-A-associated carcinogenesis. PMID- 20011138 TI - Relational memory and the hippocampus: representations and methods. AB - Since the discovery of the importance of the hippocampus for normal memory, considerable research has endeavored to characterize the precise role played by the hippocampus. Previously we have offered the relational memory theory, which posits that the hippocampus forms representations of arbitrary or accidentally occurring relations among the constituent elements of experience. In a recent report we emphasized the role of the hippocampus in all manner of relations, supporting this claim with the finding that amnesic patients with hippocampal damage were similarly impaired on probes of memory for spatial, sequential, and associative relations. In this review we place these results in the context of the broader literature, including how different kinds of relational or source information are tested, and consider the importance of specifying hippocampal function in terms of the representations it supports. PMID- 20011139 TI - Pain modulation by nitric oxide in the spinal cord. AB - Nitric oxide (NO) is a versatile messenger molecule first associated with endothelial relaxing effects. In the central nervous system (CNS), NO synthesis is primarily triggered by activation of N-methyl-D-aspartate (NMDA) receptors and has a Janus face, with both beneficial and harmful properties. There are three isoforms of the NO synthesizing enzyme nitric oxide synthase (NOS): neuronal (nNOS), endothelial (eNOS), and inducible nitric oxide synthase (iNOS), each one involved with specific events in the brain. In the CNS, nNOS is involved with modulation of synaptic transmission through long-term potentiation in several regions, including nociceptive circuits in the spinal cord. Here, we review the role played by NO on central pain sensitization. PMID- 20011140 TI - Neurophysiological mechanisms involved in auditory perceptual organization. AB - In our complex acoustic environment, we are confronted with a mixture of sounds produced by several simultaneous sources. However, we rarely perceive these sounds as incomprehensible noise. Our brain uses perceptual organization processes to independently follow the emission of each sound source over time. If the acoustic properties exploited in these processes are well-established, the neurophysiological mechanisms involved in auditory scene analysis remain unclear and have recently raised more interest. Here, we review the studies investigating these mechanisms using electrophysiological recordings from the cochlear nucleus to the auditory cortex, in animals and humans. Their findings reveal that basic mechanisms such as frequency selectivity, forward suppression and multi-second habituation shape the automatic brain responses to sounds in a way that can account for several important characteristics of perceptual organization of both simultaneous and successive sounds. One challenging question remains unresolved: how are the resulting activity patterns integrated to yield the corresponding conscious percepts? PMID- 20011141 TI - The brian simulator. AB - "Brian" is a simulator for spiking neural networks (http://www.briansimulator.org). The focus is on making the writing of simulation code as quick and easy as possible for the user, and on flexibility: new and non standard models are no more difficult to define than standard ones. This allows scientists to spend more time on the details of their models, and less on their implementation. Neuron models are defined by writing differential equations in standard mathematical notation, facilitating scientific communication. Brian is written in the Python programming language, and uses vector-based computation to allow for efficient simulations. It is particularly useful for neuroscientific modelling at the systems level, and for teaching computational neuroscience. PMID- 20011142 TI - Hebbian reverberations in emotional memory micro circuits. AB - The study of memory in most behavioral paradigms, including emotional memory paradigms, has focused on the feed forward components that underlie Hebb's first postulate, associative synaptic plasticity. Hebb's second postulate argues that activated ensembles of neurons reverberate in order to provide temporal coordination of different neural signals, and thereby facilitate coincidence detection. Recent evidence from our groups has suggested that the lateral amygdala (LA) contains recurrent microcircuits and that these may reverberate. Additionally this reverberant activity is precisely timed with latencies that would facilitate coincidence detection between cortical and sub cortical afferents to the LA. Thus, recent data at the microcircuit level in the amygdala provide some physiological evidence in support of the second Hebbian postulate. PMID- 20011143 TI - Reconstruction of virtual neural circuits in an insect brain. AB - The reconstruction of large-scale nervous systems represents a major scientific and engineering challenge in current neuroscience research that needs to be resolved in order to understand the emergent properties of such systems. We focus on insect nervous systems because they represent a good compromise between architectural simplicity and the ability to generate a rich behavioral repertoire. In insects, several sensory maps have been reconstructed so far. We provide an overview over this work including our reconstruction of population activity in the primary olfactory network, the antennal lobe. Our reconstruction approach, that also provides functional connectivity data, will be refined and extended to allow the building of larger scale neural circuits up to entire insect brains, from sensory input to motor output. PMID- 20011145 TI - Identification and characterization of merozoite surface protein 1 epitope. AB - Malaria is an important tropical infection which urgently requires intervention of an effective vaccine. Antigenic variations of the parasite and allelic diversity of the host are main problems in the development of an effective malaria vaccine. Cytotoxic T lymphocytes (CTL) directed against Plasmodium falciparum-derived antigens are shown to play an important role for the protection against malaria. The merozoite surface protein 1 (MSP1) is expressed in all the four life-cycle stages of Plasmodium falciparum and did not find any sequence similarity to human and mouse reference proteins. MSP1 is a known target of the immune response and a single CTL epitope binding to the HLA-A*0201 is available for merozoite form. Here, we report the results from the computational characterization of MSP1, precursor (1720 residue) and screening of highest scoring potential CTL epitopes for 1712 overlapping peptides binding to thirty four HLA class-I alleles and twelve HLA class-I supertypes (5 HLA-A and 7 HLA-B) using bioinformatics tools. Supertypes are the clustered groups of HLA class-I molecules, representing a sets of molecules that share largely overlapping peptide binding specificity. The prediction results for MSP1 as adhesin and adhesin-like in terms of probability is 1.0. Results also show that MSP1 has orthologs to other related species as well as having non allergenicity and single transmembrane properties demonstrating its suitability as a vaccine candidate. The predicted peptides are expected to be useful in the design of multi-epitope vaccines without compromising the human population coverage. PMID- 20011144 TI - Inducing illusory ownership of a virtual body. AB - We discuss three experiments that investigate how virtual limbs and bodies can come to feel like real limbs and bodies. The first experiment shows that an illusion of ownership of a virtual arm appearing to project out of a person's shoulder can be produced by tactile stimulation on a person's hidden real hand and synchronous stimulation on the seen virtual hand. The second shows that the illusion can be produced by synchronous movement of the person's hidden real hand and a virtual hand. The third shows that a weaker form of the illusion can be produced when a brain-computer interface is employed to move the virtual hand by means of motor imagery without any tactile stimulation. We discuss related studies that indicate that the ownership illusion may be generated for an entire body. This has important implications for the scientific understanding of body ownership and several practical applications. PMID- 20011146 TI - Structural prediction and analysis of VIH-related peptides from selected crustacean species. AB - The tentative elucidation of the 3D-structure of vitellogenesis inhibiting hormone (VIH) peptides is conversely underprivileged by difficulties in gaining enough peptide or protein, diffracting crystals, and numerous extra technical aspects. As a result, no structural information is available for VIH peptide sequences registered in the Genbank. In this situation, it is not surprising that predictive methods have achieved great interest. Here, in this study the molt inhibiting hormone (MIH) of the kuruma prawn (Marsupenaeus japonicus) is used, to predict the structure of four VIHrelated peptides in the crustacean species. The high similarity of the 3D-structures and the calculated physiochemical characteristics of these peptides suggest a common fold for the entire family. PMID- 20011147 TI - Characterization of allergenic epitopes of Ory s1 protein from Oryza sativa and its homologs. AB - Vaccination is the most effective technique suggested now days for allergy treatment. Recombinant-based approaches are mostly focused on genetic modification of allergens to produce molecules with reduced allergenic activity and conserved antigenicity. The molecules developed for vaccination in allergy possess significantly reduced allergenicity in terms of IgE binding, and therefore will not lead to anaphylactic reactions upon injection. This approach is probably feasible with every peptide allergen with known amino acid sequence. In this study an in silico approach was used to investigate allergenic protein sequences. Motif analysis of these sequences reveals the allergenic epitopes in the amino acid sequences. Physicochemical analysis of protein sequences shows that the homolog allergens of Ory s1 are highly correlated with the aromaticity, GRAVY and cysteine content. Moreover, phylogenetic analysis of Ory s1 with other sequences reveals that Oryza sativa japonica and Zea mays are close homologs, whilst Lolium perenne and Dactylis glomerata are found to be remote homologs. The multiple sequence alignment reveals of Ory s1 with all its homologs in this study reveals the high conservation of residues in DPBB_1 domain (amino acid residue positions 86- 164) and was found distinctly in all the sequences. These findings support the proposal that allergenic epitopes encompass conserved residues. The consensus allergenic was found to be mainly composed of hydrophobic residues. The functional sites of allergenic proteins reported in this study shall be attenuated to develop hypoallergenic vaccine. The sequence comparison strategy adopted in this study would pave way effective evolutionary analysis of these allergens. PMID- 20011148 TI - Mitochondrial antioxidant defence in radio-resistant Lepidopteran insect cells. AB - Cells isolated from Lepidopteran insects (butterfly and moths) display very high radioresistance as compared to mammals and other insect species. Since free radical induced mitochondrial damage under stress conditions is very crucial for cellular fate determination, antioxidant system is the major protective modality required to minimize stress-induced damage and to modulate cellular sensitivity. In this study, we predict the mitochondrial localization potential and co existence of important antioxidant enzymes in insect cells and compare with other radiosensitive (mammals, Dipteran insects) and radioresistant (nematodes) species. Our study clearly demonstrates the inter-species variation in then localization potential of various antioxidant enzymes. A higher mitochondrial localization potential as a function of mitoprot score was evident for all important antioxidant enzymes in the lepidopteran insect Bombyx mori (Mn-SOD, 0.694; GPx, 0.862; TRPx, 0.997; TR, 0.9), besides an unusual mitochondrial localization prediction for catalase (0.453). We further found coexistence of glutathione and thioredoxin system in the mitochondria of lepidopteran insects as also reported in various plant species. On the basis of above observations, we hypothesize that a strong mitochondrial antioxidant enzyme system including the unusual coexistence of catalase, glutathione and thioredoxin system may help minimize the free radical mediated damage to mitochondria and can contribute to the intrinsic radioresistance of lepidopteran insects. PMID- 20011149 TI - Insight to pyrazinamide resistance in Mycobacterium tuberculosis by molecular docking. AB - Pyrazinamide (PZA) - an important drug in the anti-tuberculosis therapy, activated by an enzyme Pyrazinamidase (PZase). The basis of PZA resistance in Mycobacterium tuberculosis was owing to mutation in pncA gene coding for PZase. Homology modeling of PZase was performed using software Discovery Studio (DS) 2.0 based on the crystal structure of the PZase from Pyrococcus horikoshii (PDB code 1im5), in this study. The model comprises of one sheet with six parallel strands and seven helices with the amino acids Asp8, Asp49, Trp68, Lys96, Ala134, Thr135 and Cys138 at the active site. Five mutants were generated with Gly at position 8, Thr at position 96, Arg at position 104, Tyr and Ser at position 138. The Wild type (WT) and five mutant models were docked with PZA. The results indicate that the mutants Lys96Thr, Ser104Arg Asp8Gly and Cys138Tyr may contribute to higher level drug resistance than Cys138Ser. These models provide the first in-silico evidence for the binding interaction of PZA with PZase and form the basis for rationalization of PZA resistance in naturally occurring pncA mutant strains of M. tuberculosis. PMID- 20011150 TI - Identification of mutations at the antigenic and glycosylation sites in hemagglutinin protein of H5N1 strain. AB - Hemagglutinin (HA) is the principal antigen, present on the viral surface. It is the primary target for neutralizing antibodies. In this paper, we have carried out studies on human hemagglutinin protein from H5N1 strain with homologous hemagglutinin from non-human sources of H5N1 strains. In all strains, part of the antigenic site (128-141) predicted by computer program "Antigenic", corresponds to immunodominant site Sa of H1 subtype. In AAF02304 strain, A156-->S156 mutation lies at the antigenic subsite of site 2 that corresponds to site B in the H3 subtype. In some strains of non-human origins, there are mutations at the antigenic sites. Interestingly, in AAY56367 strain mutation L138-->H138 lies at the receptor binding site, which also overlaps the antigenic site. Therefore, this amino acid substitution may influence both the specificity of receptor recognition and antibody binding. Seven potential glycosylation sites in human HA and in some strains of non-human sources have been predicted by computer program, Scan Prosite. In some strains of HA from non-human sources because of mutation, an additional glycosylation site appeared at the antigenic site. Therefore in these strains the oligosaccharides will mask the surface of HA as well as antigenic site. Hence these strains will not be recognized by host immune system. PMID- 20011151 TI - HIV reverse transcriptase: structural interpretation of drug resistant genetic variants from India. AB - The reverse transcriptase (RT) enzyme is the prime target of nucleoside/ nucleotide (NRTI) and non-nucleoside (NNRTI) reverse transcriptase inhibitors. Here we investigate the structural basis of effects of drug-resistance mutations in clade C RT using three-dimensional structural modeling. Apropos the expectation was for unique mechanisms in clade C based on interactions with amino acids of p66 subunit in RT molecule. 3-D structures of RT with mutations found in sequences from 2 treatment naive, 8 failed and one reference clade C have been modeled and analyzed. Models were generated by computational mutation of available crystal structures of drug bound homologous RT. Energy minimization of the models and the structural analyses were carried out using standard methods. Mutations at positions 75,101,118,190,230,238 and 318 known to confer drug resistance were investigated. Different mutations produced different effects such as alteration of geometry of the drug-binding pocket, structural changes at the site of entry of the drug (into the active site), repositioning the template bases or by discriminating the inhibitors from their natural substrates. For the mutations analyzed, NRTI resistance was mediated mainly by the ability to discriminate between inhibitors and natural substrate, whereas, NNRTI resistance affected either the drug entry or the geometry of the active site. Our analysis suggests that different mutations result in different structural effects affecting the ability of a given drug to bind to the RT. Our studies will help in the development of newer drugs taking into account the presence of these mutations and the structural basis of drug resistance. PMID- 20011152 TI - GSTaxClassifier: a genomic signature based taxonomic classifier for metagenomic data analysis. AB - GSTaxClassifier (Genomic Signature based Taxonomic Classifier) is a program for metagenomics analysis of shotgun DNA sequences. The program includes a simple but effective algorithm, a modification of the Bayesian method, to predict the most probable genomic origins of sequences at different taxonomical ranks, on the basis of genome databases;a function to generate genomic profiles of reference sequences with tri-, tetra-, penta-, and hexa-nucleotide motifs for setting a user-defined database; two different formats (tabular- and tree-based summaries) to display taxonomic predictions with improved analytical methods; and effective ways to retrieve, search, and summarize results by integrating the predictions into the NCBI tree-based taxonomic information.GSTaxClassifier takes input nucleotide sequences and using a modified Bayesian model evaluates the genomic signatures between metagenomic query sequences and reference genome databases. The simulation studies of a numerical data sets showed that GSTaxClassifier could serve as a useful program for metagenomics studies, which is freely available at http://helix2.biotech.ufl.edu:26878/metagenomics/. PMID- 20011153 TI - Genomic Target Database (GTD): a database of potential targets in human pathogenic bacteria. AB - A Genomic Target Database (GTD) has been developed having putative genomic drug targets for human bacterial pathogens. The selected pathogens are either drug resistant or vaccines are yet to be developed against them. The drug targets have been identified using subtractive genomics approaches and these are subsequently classified into Drug targets in pathogen specific unique metabolic pathways,Drug targets in host-pathogen common metabolic pathways, and Membrane localized drug targets. HTML code is used to link each target to its various properties and other available public resources. Essential resources and tools for subtractive genomic analysis, sub-cellular localization, vaccine and drug designing are also mentioned. To the best of authors knowledge, no such database (DB) is presently available that has listed metabolic pathways and membrane specific genomic drug targets based on subtractive genomics. Listed targets in GTD are readily available resource in developing drug and vaccine against the respective pathogen, its subtypes, and other family members. Currently GTD contains 58 drug targets for four pathogens. Shortly, drug targets for six more pathogens will be listed. AVAILABILITY: GTD is available at IIOAB website http://www.iioab.webs.com/GTD.htm. It can also be accessed at http://www.iioabdgd.webs.com.GTD is free for academic research and non-commercial use only. Commercial use is strictly prohibited without prior permission from IIOAB. PMID- 20011154 TI - The Influenza-A mystery: insight from Bioinformatics resources and analysis. PMID- 20011155 TI - Pinning technique for shoulder fractures in adolescents: computer modelling of percutaneous pinning of proximal humeral fractures. AB - BACKGROUND: In the technique of percutaneous pinning of proximal humerus fractures, the appropriate entry site and trajectory of pins is unknown, especially in the adolescent population. We sought to determine the ideal entry site and trajectory of pins. METHODS: We used magnetic resonance images of nonfractured shoulders in conjunction with radiographs of shoulder fractures that were treated with closed reduction and pinning to construct 3-dimensional computer-generated models. We used engineering software to determine the ideal location of pins. We also conducted a literature review. RESULTS: The nonfractured adolescent shoulder has an articular surface diameter of 41.3 mm, articular surface thickness of 17.4 mm and neck shaft angle of 36 degrees. Although adolescents and adults have relatively similar shoulder skeletal anatomy, they suffer different types of fractures. In our study, 14 of 16 adolescents suffered Salter-Harris type II fractures. The ideal location for the lateral 2 pins in an anatomically reduced shoulder fracture is 4.4 cm and 8.0 cm from the proximal part of the humeral head directed at 21.2 degrees in the coronal plane relative to the humeral shaft. CONCLUSION: Operative management of proximal humerus fractures in adolescents requires knowledge distinct from that required for adult patients. This is the first study to examine the anatomy of the nonfractured proximal humerus in adolescents. This is also the first study to attempt to model the positioning of percutaneous proximal humerus pins. PMID- 20011156 TI - Antibiotic prophylaxis for total joint replacement surgery: results of a survey of Canadian orthopedic surgeons. AB - BACKGROUND: The role of perioperative antibiotic prophylaxis in total joint replacement (TJR) surgery is well established. Whereas guidelines have been published in some countries, in Canada controversy persists concerning the best clinical practice for perioperative antibiotic prophylaxis in TJR. METHODS: We conducted a survey of 590 practising orthopedic surgeons performing TJR in Canada to assess current antibiotic prophylaxis practice. The survey included questions pertaining to antibiotic prophylaxis indications, antibiotic choice, dosing, route and timing of administration in the primary and revision arthroplasty setting, as well as postoperative wound drainage evaluation and management. RESULTS: The response rate after 2 mail-outs was 410 of 590 (69.5%). Current antibiotic prophylaxis regimens varied widely among surgeons, underscoring the controversy that exists regarding what constitutes best clinical practice. CONCLUSION: Opinions regarding use of perioperative antibiotic prophylaxis in TJR vary widely among orthopedic surgeons in Canada, illustrating the controversy as to what constitutes best clinical practice. This survey also points to a lack of consensus about the current management of postoperative wound drainage. PMID- 20011157 TI - Motorcycle-related trauma in Alberta: a sad and expensive story. AB - BACKGROUND: Trauma caused by motorcycle-related injuries is extensive, expensive and increasing. Recent American literature reported that in 2004 the chance of a motorcyclist dying was 34 times greater than that for someone using any other motor vehicle for every mile travelled. In the United Kingdom a motorcyclist is killed or seriously injured every 665,894 km, compared with 18,661,626 km for cars. If this pattern is repeated in Canada, then this information should be in the public domain to support initiatives for injury prevention. METHODS: We gathered and analyzed retrospective population data on the injury patterns of adult motorcyclists and other adult motor vehicle drivers and passengers across Alberta from Apr. 1, 1995, to Mar. 31, 2006. We collected data from 3 Alberta sources: the Alberta Trauma Registry, the Alberta Office of the Chief Medical Examiners and the Government of Alberta Department of Infrastructure and Transportation. We compared the numbers and causes of crashes, injuries and deaths, as well as the acute care costs on the roads, and specifically compared motorcycle-related injuries to all other motor vehicle-related injuries. RESULTS: There were 70,605 registered motorcycles and 2,748,204 other registered motor vehicles in Alberta during the study period. During these 11 years, there were 286 motorcyclists killed and 712 were severely injured, representing a total of 998 injuries and deaths. There was 5386 deaths related to other motor vehicles and 6239 severe injuries, for a total of 11,625 injuries and deaths. This represents a percentage of 1.4% of all registered motorcycles and 0.4% of all other registered motor vehicles (3.5 times more motorcyclist injuries). The impact on the health care system can be measured in several ways. During the period of this study, motorcyclists accounted for 10,760 bed days. Assuming the patient was not admitted to intensive care, each admission cost Can$9200 (average in 2008). CONCLUSION: Analysis of the data shows that motorcyclists are more than 3.5 times more likely to get injured or die than other motor vehicle drivers. All of the injuries in motorcyclists occurred during the summer months, leading to an adjusted risk of almost 8 times compared with that of the motor vehicle driver. PMID- 20011158 TI - Outcomes of simultaneous and delayed resections of synchronous colorectal liver metastases. AB - BACKGROUND: The optimal strategy for the treatment of synchronous colorectal liver metastases has not been established yet. In this study, we present the outcomes and survival rates of the patients who underwent simultaneous or delayed resections. METHODS: We performed a retrospective analysis of liver resections in our institution between 1997 and 2006. RESULTS: Among the 89 patients presenting with synchronous colorectal liver metastases, 28 underwent simultaneous and 61 underwent delayed resection. Age, sex and localization of the primary tumour were similar in the 2 groups. Duration of surgery and hospital stay were longer in the simultaneous resection group, and blood loss was also greater in this group. However, these factors did not influence the frequency of complications, which did not differ between the groups. When we included data from initial colectomy, these differences were either not significant or in favour of synchronous resection. In the delayed resection group, colon resection was performed in different hospitals. The 1-, 3- and 5-year survival rates were 78%, 70% and 45%, respectively, in the simultaneous and 88%, 55% and 38%, respectively, in the delayed resection groups. CONCLUSION: In select patients, the risk of simultaneous resection of synchronous colorectal liver metastases is comparable to delayed resection, and increases in blood loss and operating time associated with simultaneous resections do not have a negative influence on long-term outcome. Positive outcomes of simultaneous liver resections in our study could be a result of good patient selection or experience with oncological liver surgery. PMID- 20011159 TI - Mammary ductoscopy in the evaluation and treatment of pathologic nipple discharge: a Canadian experience. AB - BACKGROUND: Mammary ductoscopy allows direct visualization of ductal epithelium using a fibreoptic microendoscope. As the first centre in Canada to apply ductoscopy to surgical practice, we report our experience with this technology. METHODS: Between 2004 and 2008, 65 women with pathologic nipple discharge underwent ductoscopy before surgical duct excision under general anesthetic. Prospective data collection included cannulation and complication rates, procedure length and lesion visualization rate compared with preoperative ductography, if performed. In addition, we classified the endoscopic appearance according to Makita and colleagues and correlated it with surgical pathology. RESULTS: It took longer than 6 months to overcome technical problems before the routine use of ductoscopy in the operating room. The ductoscope was easy to use: we achieved cannulation in 63 of 66 breast ducts (95%) and we visualized a lesion in 52 of 63 breast ducts (83%). The mean procedure length was 5.1 minutes, with no complications. Lesions seen on ductography were seen endoscopically 30 of 33 (91%) times. All 3 malignancies were seen: invasive carcinoma in 1 of 62 (1.6%) and in situ disease in 2 of 62 (3.2%) patients. Surgeons found ductoscopy helpful in defining the extent of duct excision. Except for the "polypoid solitary" class, which accurately predicted a papilloma (23/23), we found poor correlation between Makita and colleague's endoscopic classification and final pathology. CONCLUSION: Ductoscopy is feasible, safe and practical. Our surgeons routinely use it to identify the location and extent of duct excision without ordering preoperative ductography. Identifying pathology based on the endoscopic appearance is unreliable unless the lesion is solitary and polypoid. PMID- 20011160 TI - Five-year outcomes of laparoscopic adjustable gastric banding and laparoscopic Roux-en-Y gastric bypass in a comprehensive bariatric surgery program in Canada. AB - BACKGROUND: Bariatric surgery remains the most effective modality to induce sustainable weight loss in the morbidly obese. Our aim was to compare outcomes between the laparoscopic Roux-en-Y gastric bypass (LRYGBP) and the laparoscopic adjustable gastric banding device (LAGBD) method with 5-year follow-up in a Canadian bariatric surgery centre. METHODS: This is a retrospective outcomes analysis of 1035 laparoscopic bariatric procedures performed over 7 years. We extracted data from our prospectively collected bariatric surgery registry from Feb. 1, 2002, to Jun. 30, 2008. We evaluated patient demographics, weight loss, complications, mortality and need for revision surgery by procedure type. RESULTS: We examined outcomes in 149 (14.4%) LAGBD and 886 (85.6%) LRYGBP procedures. The mean body mass index (BMI) was significantly higher in the LRYGBP group (50.9, standard deviation [SD] 8.9, v. 45.0, SD 6.7) whereas age and sex ratio were the same. There were 3 deaths (0.3%) in the LRYGBP group and no deaths in the LAGBD group. Sixteen patients (10.8%) in the LAGBD group needed conversion to LRYGBP because of poor weight loss, band intolerance, band erosion or slippage, and 6 patients (0.7%) in the LRYGBP group required revision because of inability to achieve the desired weight loss. The percent excess-weight loss was 41, 49, 59, 60 and 61 at 1, 2, 3, 4 and 5 years postsurgery for the LAGBD patients who kept their band, and 70, 79, 79, 79 and 75 for the LRYGBP patients. CONCLUSION: Laparoscopic weight loss surgery can be performed safely with acceptable mortality. Our study suggests superior weight loss and low revision requirement for the LRYGBP, making this a more durable procedure in a publicly funded health care system. PMID- 20011161 TI - Translation of the Patient Scar Assessment Scale (PSAS) to French with cross cultural adaptation, reliability evaluation and validation. AB - BACKGROUND: Patient esthetic satisfaction related to scarring after orthopedic surgery was rarely assessed before the development of the Patient Scar Assessment Scale (PSAS). The purpose of our study was to translate and validate the PSAS and assess the psychometric properties of the French version. METHODS: We conducted a staged validation with forward and backward translation and concurrent validation. The validation committee comrpised bilingual experts. The patient validation sample comprised 53 orthopedic surgery patients who were assessed at a minimum of 1 year postoperatively. We followed a standardized process for cross cultural adaptation to develop and assess the French version. First, 2 independent translators completed the forward translation of the PSAS and then met to achieve a consensus version. This consolidated version was then backward translated into English and cross-verified with the original version. A group of orthopedic and plastic surgeons assessed this version for content validity. We assessed the test-retest reliability of the new French scale, which was filled out twice by a cohort of 53 patients, using scale distribution analysis, internal consistency (Chronbach alpha) and absolute agreement (intraclass correlation coefficients [ICC (2,1)]). RESULTS: The level of agreement on the translation process between the translators initially and then later among the expert panel was high. The reliability of the translated version (PSAS-Fr) and its internal consistency was high (Chronback alpha 0.87-0.98 for each of the 6 questions), and the test-retest reliability was excellent (ICC 0.96). On the other hand, there was no bias between occasions (retests difference -0.24) and the scores fell within 2 standard deviations of 5. Older patients had higher satisfaction about scar appearance. CONCLUSION: The PSAS-Fr was successfully translated from the original English version and demonstrated strong cross-sectional psychometric properties. Further assessment in longitudinal studies is warranted. PMID- 20011162 TI - Hepatic resection in Canada: rates and geographic variation. AB - BACKGROUND: Liver resection (LR) is the only curative therapy for hepatic malignancy. Despite this, the rates of LR across Canada are unknown. We sought to determine the rate and regional variation of LR in Canada, its provinces and census divisions for the period 1995-2004. METHODS: We obtained discharge data for 1995-2004 from all provinces but Quebec from the Canadian Institute for Health Information, and we searched for LR by residential site. Then we calculated rates and variations in rates. RESULTS: The national age- and sex adjusted rate of LR per 100,000 people aged 18 years and older increased from 3.22 in 1995 to 5.86 in 2004. There was a high degree of variation across provinces and census divisions. CONCLUSION: Liver resection volumes are increasing in Canada. Significant regional variation exists for LR in Canada, likely reflecting a variety of important supply-demand issues. PMID- 20011163 TI - Can Roux-en-Y gastric bypass provide a lifelong solution for diabetes mellitus? AB - BACKGROUND: The surgical treatment of diabetes had witnessed progressive development and success since the first case of pancreatic transplantation. Although this was a great step, wide clinical application was limited by several factors. Bariatric surgery such as gastric bypass is emerging as a promising option in obese patients with type 2 diabetes. The aim of this article is to explore the current application of gastric bypass in patients with type 2 diabetes and the theoretical bases of gastric bypass as a treatment option for type 1 diabetes. METHODS: We performed a MEDLINE search for articles published from August 1955 to December 2008 using the words "surgical treatment of diabetes," "etiology of diabetes" and "gastric bypass." RESULTS: We identified 3215 studies and selected 72 relevant papers for review. Surgical treatment of diabetes is evolving from complex pancreatic and islets transplantation surgery for type 1 diabetes with critical postoperative outcome and follow-up to a metabolic surgery, including gastric bypass. Gastric bypass (no immune suppression or graft rejection) has proven to be highly effective treatment for obese patients and nonobese animals with type 2 diabetes. There are certain shared criteria between types 1 and 2 diabetes, making a selected spectrum of the disease a potential target for metabolic surgery to improve or cure diabetes. CONCLUSION: Roux-en-Y gastric bypass is a promising option for lifelong treatment of type 2 diabetes. It has the potential to improve or cure a selected spectrum of type 1 diabetes when performed early in the disease. Further animal model studies or randomized controlled trials are needed to support our conclusion. PMID- 20011164 TI - Occlusion of the common and internal iliac arteries for aortoiliac aneurysm repair: experience with the Amplatzer vascular plug. AB - BACKGROUND: We sought to evaluate and describe our centre's experience with the Amplatzer vascular plug (AVP) for the occlusion of common and internal iliac arteries (CIA; IIA) during endovascular aortic aneurysm repair (EVAR). METHODS: We performed a retrospective analysis of 20 consecutive patients between October 2006 and December 2007, who underwent occlusion of the CIA or IIA before or during EVAR to prevent endoleak. RESULTS: Among these 20 patients, 21 occlusion procedures occurred and 20 were successful. In the only unsuccessful case, the patient had EVAR, but occlusion with an AVP was not possible because of severe narrowing at the origin of the vessel. Of the successfully treated patients, 2 presented with ruptured aneurysms, whereas the others had elective procedures. Eleven patients received aortouni-iliac grafts and femoral-femoral bypass, and 9 patients received a bifurcated stent graft. In 5 patients, the AVP occlusion and EVAR procedures were staged; in these cases occlusion occurred first, followed by EVAR on average 29 (standard deviation [SD] 23) days later. We deployed 7 AVPs in the CIA, whereas 13 were deployed in the IIA. The average diameter of the vessels occluded was 10 (SD 1) mm and the average size of the device used was 13 (SD 1) mm, representing a device diameter 28% (SD 2%) greater than the vessel diameter. We used a single device in 18 patients, whereas 2 devices were deployed in the same artery in 2 patients. Four patients underwent concomitant coil embolization. On follow-up computed tomography (CT) scans, all occlusion procedures were clinically successful. At the 14-month (SD 1 mo) follow-up, 4 patients had a small type-II endoleak unrelated to the occlusion procedure and 1 had a type-I endoleak that required graft limb extension. Four patients had buttock claudication but none had changes in sexual function, ischemic complications or device dislodgement on CT scans. CONCLUSION: The AVP is a safe and effective method to occlude the CIA and IIA in patients undergoing EVAR. PMID- 20011165 TI - Adrenal incidentalomas in the laparoscopic era and the role of correct surgical indications: observations from 255 consecutive adrenalectomies in an Italian series. AB - BACKGROUND: The purpose of our study was to evaluate the impact of laparoscopic adrenalectomy on patients with incidentalomas. We analyzed the results of a multi centre trial that was performed to evaluate the effectiveness of imaging (computed tomography and magnetic resonance imaging) to obtain a correct preoperative diagnosis. METHODS: We obtained our data from the results of a questionnaire that was distributed by mail or email in May 2005 to several surgical units operating in the Campania Region, Italy. Lap Club, a collaborative laparoscopic surgery study group founded in Naples in 1995, distributed the questionnaire. Thirteen centres participated in the audit. In all, we analyzed 255 adrenalectomies performed on 250 patients. We performed statistical analysis using SPSS software. RESULTS: The distribution of pathologic findings demonstrates that the number of lesions caused by cancer discovered from a preoperative indication of incidentaloma has been even smaller (1/114, 0.8%) than the previous numbers reported in the literature. Moreover, whereas most patients with adrenal cancer had lesions larger than 6 cm (7/8, 87.5%), the majority of patients with adrenal metastases had lesions 6 cm or smaller (10/12, 83.3%). Different indications for adrenalectomy emerged on comparison of endocrine surgery units with general surgery units. This difference appears to be significant (p < 0.001), especially on evaluation of the number of nonfunctioning adenomas and the number of endocrine lesions that were observed and treated. CONCLUSION: Laparoscopy remains the gold standard method for adrenalectomy, but its availability must not obligate physicians to treat with surgery when an incidentaloma is detected through imaging. Adrenal malignancies when metastatic are often 6 cm or smaller. If they are single and they originated from a non small lung cancer, they must be removed. The endocrine surgery unit remains the best setting to evaluate and treat adrenal gland surgical pathology. PMID- 20011166 TI - Cystic lymphangioma of the jejunal mesentery presenting with acute abdomen in an adult. PMID- 20011168 TI - Renal artery avulsion from blunt abdominal trauma in a horseshoe kidney: endovascular management and an unexpected complication. PMID- 20011167 TI - Primary solid neuroendocrine carcinoma of the breast. PMID- 20011169 TI - Thoracic splenosis accompanied by diaphragmatic hernia. PMID- 20011170 TI - An unexpected severe complication after a negative laparoscopic appendectomy. PMID- 20011171 TI - Hemocholecyst associated with antithrombotic therapy. PMID- 20011172 TI - Jejunal carcinoma in a patient with Peutz-Jeghers syndrome. PMID- 20011173 TI - Video-assisted thoracoscopic surgery (VATS) for penetrating chest wound: thoracoscopic exploration and removal of a penetrating foreign body. PMID- 20011174 TI - Huge cystic lymphangioma of the pancreas. PMID- 20011175 TI - Successful palliation of diarrhea owing to malignant duodenocolic fistula by octreotide. PMID- 20011176 TI - Primary malignant melanoma of the tongue. PMID- 20011177 TI - Hemi- versus total hip arthroplasty for femoral neck fixation. PMID- 20011178 TI - Emphasis on prevention. PMID- 20011180 TI - The adoption of laparoscopic colorectal surgery: a national survey of general surgeons. AB - BACKGROUND: Laparoscopic surgery may become the standard of care for the treatment of colorectal disease. Little is known regarding North American patterns of practice or the limiting factors and strategies for adoption among surgeons. METHODS: We sent a 28-item questionnaire to all general surgeon members of the Royal College of Physicians and Surgeons of Canada. We derived descriptive and correlative information using chi(2), Wilcoxon rank sum and Student t tests and multivariate logistic regression. RESULTS: The return rate was 55% (694/1266). A total of 67% (462/694; 95% confidence interval 63%-70%) of respondents perform colorectal surgery. Of these, 54% perform laparoscopic colorectal surgery. Multivariate logistic regression identified 5 factors related to performing laparoscopic colorectal surgery: fewer years in practice (p < 0.001), male sex (p = 0.015), practising in the province of Quebec (p = 0.005), university-hospital affiliation (p = 0.034) and minimally invasive surgery fellowship training (p = 0.023). Lack of adequate operating time and formal training were the main reasons cited by surgeons not offering laparoscopic colon resections. Most surgeons (67%) felt that site visits from a minimally invasive surgeon would represent the most effective training method for acquiring advanced laparoscopic skills. CONCLUSION: About half of Canadian general surgeons offer laparoscopic colorectal resections. Recent graduation, male sex, practice location, university-hospital affiliation and minimally invasive surgery training are significant predictors for offering a laparoscopic approach. Lack of operative time and formal training are the main barriers to adoption of the technique. Site visits by trained laparoscopic surgeons is the preferred method of acquiring advanced skills. PMID- 20011181 TI - Conversion in laparoscopic cholecystectomy after gastric resection: a 15-year review. AB - BACKGROUND: Gastrectomy or truncal vagotomy is known to increase the incidence of cholelithiasis. Many of these patients will become symptomatic, and the adhesions from their gastric resection may make laparoscopic cholecystectomy much more difficult. METHODS: We prospectively assessed the data for the 15-year cumulative laparoscopic cholecystectomy experience of 1 surgeon at a university teaching hospital with respect to conversion and postoperative outcomes, with particular attention to patients having had previous gastric resections. RESULTS: Patients with previous gastrectomies had similar operative times (mean 81.1, range 45-120 min), a higher conversion rate (64.2%) and a higher complication rate (35.7%) than those who had had other previous upper abdominal surgeries (mean 73.2, range 35-130 min, conversion 25% and complication 11.3%) and those without previous abdominal surgeries (mean 66.5, range 25-250 min, conversion 2.7% and complication 4.5%). CONCLUSION: Preoperative knowledge of the increased conversion rate and increased morbidity will inform surgical planning for both the surgeon and the patient. PMID- 20011182 TI - Safety and hospital costs of Achilles tendon surgery: the serendipitous impact of a randomized clinical trial. AB - BACKGROUND: Surgical management is currently the most common treatment offered for ruptured Achilles tendon; however, the length of hospital stay varies among hospitals. The objective of our study was to compare the costs associated with hospital stay and safety in 2 groups of patients undergoing surgical treatment of Achilles tendon ruptures. The first group of patients represented the surgical arm of a randomized clinical trial, the Multicentre Achilles Tendon Treatment Study (MATTS), and the second group comprised surgically treated patients not included in that trial (non-MATTS). METHODS: We performed an observational analytical retrospective chart review of all patients surgically treated for acute Achilles tendon ruptures at all Calgary-area hospitals over a 3-year period (October 2002-September 2005) who met our inclusion criteria. We determined the costs associated with hospital stay, complications and readmissions for all patients. RESULTS: A total of 282 patients met our inclusion criteria. Hospital admission costs of patients enrolled in the MATTS trial were less than those of non-MATTS patients treated at each of the 3 Calgary adult hospitals. Non-MATTS patients remained in hospital for 1-4 days in contrast to MATTS patients, who were typically admitted and discharged the day of surgery. The complication rate was similar in both groups (MATTS 9% v. non-MATTS 8%). There were no readmissions in the MATTS group and 2 in the non-MATTS group. Between October 2002 and September 2005, an additional $236,436 was spent on non-MATTS patients who underwent surgery for Achilles tendon rupture. CONCLUSION: Acute achilles tendon repair surgery is safe and less costly as an out-patient procedure. PMID- 20011183 TI - Potential triaging of referrals for lumbar spinal surgery consultation: a comparison of referral accuracy from pain specialists, findings from advanced imaging and a 3-item questionnaire. AB - BACKGROUND: Waiting times to see a spinal surgeon are among the highest in Canada. However, most patients who are referred would not benefit from surgical care. Effective triaging of surgical candidates may reduce morbidity related to prolonged waiting times and optimize use of limited resources. METHODS: We administered a questionnaire consisting of 3 items identifying leg-dominant or back-dominant pain among 119 consecutive patients who presented at a community spinal pain centre or a spinal surgical unit for assessment of an elective lumbar problem. We analyzed the questionnaire under 2 different scenarios: 1 hypothesized to be more sensitive and 1 hypothesized to be more specific. RESULTS: For the "sensitive" scenario of clearly back-dominant pain, the sensitivity of the questionnaire was 100% in identifying appropriate surgical candidates. For the "specific" scenario of leg-dominant pain, the questionnaire had a sensitivity of 83% and specificity of 73% in identifying appropriate surgical candidates, which was significantly superior to findings on computed tomography or magnetic resonance imaging (i.e., presence of neurocompressive lesions). When comparing the accuracy of the questionnaire in identifying appropriate surgical candidates to that of an assessment performed by a pain specialist at an acute spinal pain clinic, we found no statistically significant differences between the 2 methods. CONCLUSION: Use of the questionnaire when triaging patients may decrease the number of unnecessary referrals to spine surgeons. Adopting such a method of triaging could reduce waiting times for appropriate surgical candidates and potentially improve the outcomes of any resulting spinal surgery performed in a timely fashion. PMID- 20011184 TI - Factors influencing antibiotic prophylaxis for surgical site infection prevention in general surgery: a review of the literature. AB - BACKGROUND: Surgical site infections (SSIs) are common surgical complications that can be prevented with antibiotic prophylaxis. Research shows poor adherence to guidelines for this practice. We conducted a scoping review to identify factors or interventions that influence antibiotic prophylaxis administration. METHODS: An investigator with informatics training searched indexed (MEDLINE, Cochrane Library) and nonindexed (Internet) sources from January 1996 to February 2007. Selected studies were English-language, peer-reviewed, quantitative or qualitative studies describing factors or interventions influencing adherence to SSI-prevention guidelines or SSI rates in general surgery. Two investigators independently reviewed citations and full-text articles and extracted data, and met to compare selections or data and resolve differences through discussion. We extracted data on type of surgery, study design, intervention or factors examined and key findings. We then examined the quantity and type of studies and their findings. RESULTS: Nineteen of 192 studies met the eligibility criteria. Seven studies investigated predictors of appropriate antibiotic use through descriptive or exploratory means. Twelve evaluated adherence to antibiotic prophylaxis recommendations by comparing patient cohorts before and after the introduction of quality-improvement strategies. Individual knowledge, attitudes, beliefs and practice; team communication and allocation of responsibilities; and institutional support for promoting and monitoring practice appear to influence practice. CONCLUSION: Larger and multisite studies included in our review favour implementation of multidisciplinary pathways, individualized performance data and written or computerized order sets as quality-improvement strategies, but further research is warranted to more rigorously evaluate the effectiveness of these strategies on antibiotic prophylaxis practice. PMID- 20011185 TI - Effect of hand packing versus cement gun pressurization on cement mantle in total knee arthroplasty. AB - BACKGROUND: Gun pressurization in total knee arthroplasty (TKA) may result in better cement penetration than hand packing, leading to fewer tibial plate failures. We compared cement intrusion characteristics between vacuum mixing and gun pressurization versus hand mixing and packing in the proximal tibia among patients undergoing TKA. METHODS: We analyzed 6-week radiographs from 77 consecutive patients for cement area and zone-specific intrusion using computer assisted image analysis. RESULTS: Penetration into tibial anteroposterior zones 1 6 was not significantly different between the techniques. Intrusion depths in anteroposterior zone 7 and lateral zone 2 were significantly increased with gun pressurization, but this increase was associated with significantly longer operating room and tourniquet times. CONCLUSION: We identified no obvious advantage of vacuum mixing with gun pressurization, suggesting that continued use of the hand-packing technique may be warranted. Additional long-term failure studies must be completed to compare these techniques. PMID- 20011186 TI - Impact of orthopedic trauma consolidation on resident education. AB - BACKGROUND: In July 2005, the Saskatoon Health Region, in conjunction with its orthopedic surgeons, consolidated its management of orthopedic emergencies from 3 sites to 1, resulting in trauma patients being directed to the university hospital site (Royal University Hospital; RUH) where orthopedic residents were the first responders. We sought to measure resident workload in the emergency department, operating room and ward before and after consolidation and to measure the perceptions of residents and faculty on the newly established orthopedic trauma service. METHODS: We compared orthopedic volumes at RUH in the emergency department (ED) and trauma-related operating rooms over 2 periods of 3 months' duration before and after trauma consolidation. We developed and disseminated questionnaires evaluating issues relating to patient care; resident education, including all CanMEDS domains; resident well-being; and the orthopedic trauma service to all orthopedic residents and faculty members. RESULTS: The number of patients seen by orthopedic residents in the ED increased by 67%, the number admitted through the ED to the ward increased by 66%, and the total number of inpatients and consultations increased by 43% after the consolidation. The number of patients processed through the orthopedic emergency operating room increased by 90%. In general, response to the change was positive and appreciated by residents and faculty members. CONCLUSION: Sudden substantial increases in the volume of patients seen by orthopedic residents may not prompt negative resident responses when the overall gains offset, if not exceed, the perceived losses. PMID- 20011187 TI - Canadian Association of University Surgeons annual symposium: Continuity of care: Toronto, Ontario, Sep. 6, 2007. AB - This 2007 symposium of the Canadian Association of University Surgeons brought together surgeons from a number of jurisdictions to discuss the challenges and opportunities that reduced physician work hours will bring to the care of the surgical patient. Dr. Brian Taylor, president of the association, underscored the need to find a balance between the benefits of diminished workloads/work hours and the loss of continuity of care. He opined that Canada needs to learn from our European colleagues' experience. Dr. Per-Olof Nystrom, professor of surgery, presented the modern Swedish model of surgical care, which had to be developed as a consequence of the European Union's legal restrictions on the amount of time an individual surgeon may work. Sweden employs a team-based shared-care model driven by the individual surgeon's expertise rather than the "village factory" model of the multiskilled, multitasking approach of surgical care more prevalent in Canada. Dr. Chris de Gara, secretary treasurer of the association, presented the evidence base for (and against) work-hour restrictions and how well-designed systems can ensure effective continuity of care. Dr. Stewart Hamilton illustrated how one such system for the delivery of the emergency general surgical services has evolved at the University of Alberta Hospital, which demonstrated its effectiveness in providing quality surgical continuity of care. Dr. Debrah Wirtzfeld underscored the importance of trainee lifestyle and how modern Web based technologies can ensure reduced errors with the implementation of a "sign out" system. PMID- 20011188 TI - Choledochal cysts: part 2 of 3: Diagnosis. AB - Much about the etiology, pathophysiology, natural course and optimal treatment of cystic disease of the biliary tree remains under debate. Gastroenterologists, surgeons and radiologists alike still strive to optimize their roles in the management of choledochal cysts. To that end, much has been written about this disease entity, and the purpose of this 3-part review is to organize the available literature and present the various theories currently argued by the experts. In part 2, we explore the details surrounding diagnosis, describing the presentation and imaging of the disease. PMID- 20011189 TI - CAGS and ACS evidence based reviews in surgery. 31. The use of intensive insulin therapy and pentastarch resuscitation in patients with severe sepsis. PMID- 20011190 TI - How to work with a subgroup analysis. PMID- 20011191 TI - A simple home-made self-retaining retractor for thyroidectomy. PMID- 20011195 TI - Multidisciplinary teams in the management of rectal cancer. AB - A myriad of advances in the treatment of rectal cancer have been achieved over the last few decades. The introduction of total mesorectal excision (TME) has resulted in significant improvements in local recurrence. Surgical education on the technique has made it the standard of care. Radiation and chemotherapy combined with TME have improved results even further with stage II and III cancers. Sphincter-sparing techniques, reservoir procedures, local treatment advances, minimally invasive techniques, surgery for metastatic disease, newer chemotherapies, and extended resections for locally advanced and recurrent lesions, have all benefited the patient with rectal cancer. The goal and responsibility of colorectal surgeons treating rectal cancer patients is to understand and coordinate the wide variety of modalities available to optimize survival, minimize morbidity, and maximize quality of life for those with this difficult problem. Coordination of specialists in this time of evolution in rectal cancer treatment becomes more important than ever. Here the authors briefly review the role of the multidisciplinary team, discuss a model multidisciplinary team approach and look at evidence supporting team use as we begin this issue devoted to the multidisciplinary management of rectal cancer. PMID- 20011196 TI - Rectal cancer staging. AB - Rectal cancer staging provides critical information concerning the extent of the disease. The information gained from staging is used to determine prognosis, to guide management, and to assess response to therapy. Accurate staging is essential for directing the multidisciplinary approach to therapy. This article focuses on the evolution of staging systems, the rational for staging, and current methods used to stage rectal cancer. PMID- 20011197 TI - The pathologist's role in rectal cancer patient assessments. AB - Since Cuthbert Dukes fundamental work linking cancer stage to prognosis, the pathologist has had an ever-expanding role in the multidisciplinary management of rectal cancer. Gross dissection techniques and histologic evaluation are reviewed. The evolving tumor size, node status, metastasis classification (TNM) staging system is outlined. The pathologist's critical role in correctly assessing circumferential margins and establishing resection adequacy and staging accuracy via lymph node assessment is emphasized. PMID- 20011198 TI - Adjuvant therapy for rectal cancer. AB - Patients with stage II and III rectal cancer benefit from a multidisciplinary approach to treatment. Studies of postoperative adjuvant therapy consistently demonstrate decreases in locoregional recurrence with the use of radiation therapy. The use of postoperative chemotherapy results in improved disease-free survival and overall survival in certain studies. Preoperative radiation therapy decreases locoregional recurrence and in one study demonstrated an improvement in survival. The addition of chemotherapy to preoperative radiation results in improved locoregional control, but not survival. Preoperative chemoradiation is the standard of care for patients with clinical stage II and III rectal cancer in the United States due to improved local recurrence, acute and late toxicity, and sphincter preservation compared with postoperative chemoradiation. Promising approaches include the incorporation of new chemotherapeutic and biologic agents into chemoradiation and adjuvant chemotherapy regimens; new radiation techniques, such as the use of intraoperative radiation therapy and an accelerated concomitant radiation boost; and gene and protein expression profiling, to better predict response to treatment and prognosis. PMID- 20011199 TI - Local treatment for rectal cancer. AB - The treatment of rectal cancer includes both radical resection and local therapy. Radical resection remains the standard treatment, but is associated with increased morbidity and mortality, as well as the potential need for a temporary and occasionally, a permanent ostomy. The benefits of local treatment include a less invasive procedure with maintenance of bowel function and avoidance of a stoma. However, the efficacy of local treatment is now being challenged as the rates of recurrence after local excision alone appear to be much higher than previously thought. Although the primary goal of an oncologic resection is disease eradication, each case must be individualized to determine an optimal care plan. PMID- 20011200 TI - Total mesorectal excision: what are we doing? AB - The introduction of total mesorectal excision (TME) for rectal cancer has reduced local recurrence rates and improved oncologic outcomes, although complication rates such as anastomotic leak have also been a consequence. With the advent of neoadjuvant therapy for rectal cancer, many are questioning how this development may change the role of TME. This review presents a history of how TME evolved and a description of this technique. Complication rates, the impact of neoadjuvant therapy on local recurrence, variations of TME such as nerve-sparing proctectomy and cancer-specific mesorectal excision, and a review of functional outcomes for various methods of reconstruction are presented. PMID- 20011201 TI - Sphincter-sparing resection for rectal cancer. AB - Although there is still a place for abdominoperineal resection in the treatment of rectal cancer, the state of the art is sphincter-preserving resection. Even for the lowest of rectal cancers, using a combination of neoadjuvant chemo/radiation, total mesorectal excision, and intersphincteric proctectomy and colonic J-pouch to anal anastomosis, sphincter preservation can be achieved for most patients. The key concept in pushing sphincter preservation forward has been the realization that the deep, circumferential, or lateral margin is all important. Unless the rectal tumor involves the external sphincter muscle, there is no oncologic need to remove it, and following resection of the tumor, gastrointestinal tract continuity can be restored. PMID- 20011202 TI - Abdominoperineal resection: how is it done and what are the results? AB - Abdominoperineal resection (APR) for many years was the treatment of choice for most patients with rectal cancer. Recent advances in surgical technique and other treatment modalities have led to a marked increase in the rate of sphincter sparing operations, with a concomitant decrease in APR. However, it is still necessary in selected patients, especially those with very distal tumors or poor sphincter function. This review will cover the history of APR, current operative strategy and complications, oncologic and quality of life results, as well as potential future advances. PMID- 20011204 TI - Defining the volume-quality debate: is it the surgeon, the center, or the training? AB - The quality movement in health care is ubiquitous in our society. The volume quality debate is a central component of this that affects surgeons. In colorectal surgery and other fields, studies have demonstrated improved outcomes for patients having care provided at higher volume centers. What is unclear about this relationship however, is whether this improvement is related to the center, the surgeon, or the surgeon's training and experience. Some studies have tried to better examine this relationship and have suggested that limitations in administrative data may exaggerate the impact of a high-volume center. The use of crude mortality as the primary outcome instead of more specific outcomes such as cancer recurrence, inadequate risk data, and the failure to account for clustering of cases are other important limitations. Although higher volume likely equates to higher quality in some form, this may be more related to surgeon-specific factors rather than high-volume centers alone. The role of subspecialization, especially colorectal-trained surgeons with a high individual case volume may be the most important predictor of higher quality in colorectal surgery. This relationship may be especially important for the treatment of rectal cancer. The relationship of volume to outcomes is difficult to understand, and to appropriately answer these questions will require the collection and analysis of comprehensive, risk-adjusted data after adequate outcome measures are defined. This will only occur with significant institutional support, and a commitment to follow outcomes longitudinally and implement necessary changes to improve outcomes. PMID- 20011203 TI - Reconstruction techniques after proctectomy: what's the best? AB - There are approximately 40,000 new rectal cancer cases diagnosed each year in the United States, representing the second most common gastrointestinal malignancy (behind colon cancer). With the advent of sphincter preserving techniques, patients with mid and low colorectal cancers enjoy the benefits of better postoperative functional outcomes and quality of life; however, controversy exists over which reconstructive technique is superior in restoring bowel continuity. Construction of a straight coloanal anastomosis is technically simpler, but functional outcomes are inferior compared with colonic reservoirs. The purpose of this review is to summarize the current data regarding reconstructive techniques following proctectomy. PMID- 20011205 TI - Laparoscopic approaches to rectal cancer. AB - Laparoscopic colectomy has been proven oncologically equivalent to conventional surgery and is now generally agreed to offer patients a reduced length of stay, shorter recovery times, and improved cosmesis. In contrast, acceptance of laparoscopic proctectomy for rectal cancer has been much delayed and the enthusiasm of early studies has met considerable skepticism. For rectal cancer, it has been demonstrated that there is considerable variation between surgeons in disease-free survival and local pelvic recurrence after open proctectomy for rectal cancer. These differences are likely to be magnified when the technical challenge of laparoscopy is added to proctectomy. Minimally invasive approaches to rectal cancer need to demonstrate equivalent oncologic outcomes and maintenance or improvement in quality of life. This review will outline the current evidence for laparoscopy as a treatment option for patients with rectal cancer, emphasize the need for standardized approaches among multidisciplinary teams, and highlight the technical details of different laparoscopic operations for rectal cancer. PMID- 20011206 TI - The role of postoperative surveillance in colorectal cancer. AB - Postoperative surveillance for recurrent and/or metachronous disease is an important component of the treatment of patients with colorectal cancer. The optimal schedule of follow-up investigations remains controversial. Several randomized trials have suggested a moderate improvement in 5-year survival and earlier detection of cancer recurrence with the implementation of intensive surveillance protocols. Whether these protocols are cost-effective has yet to be determined. Current guidelines from the American Society of Colon and Rectal Surgeons recommend periodic patient follow-up with office visits, carcinoembryonic antigen (CEA) measurement, and endoscopy following potentially curative resection of colorectal cancer. PMID- 20011207 TI - Management of locally advanced primary and recurrent rectal cancer. AB - Treatment for patients with locally advanced and recurrent rectal cancer differs significantly from patients with rectal cancer restricted to the mesorectum. Adequate preoperative imaging of the pelvis is therefore important to identify those patients who are candidates for multimodality treatment, including preoperative chemoradiation protocols, intraoperative radiotherapy, and extended surgical resections. Much effort should be made to select patients with these advanced tumors for treatment in specialized referral centers. This has been shown to reduce morbidity and mortality and improve long-term survival rates. In this article, we review the best treatment options for patients with locally advanced and recurrent rectal cancer. We also emphasize the necessity of a multidisciplinary team, including a radiologist, radiation oncologist, urologist, surgical oncologist, plastic surgeon, and gynecologist in the diagnosis and treatment of patients with these pelvic tumors. PMID- 20011210 TI - From evidence to practice: consensus in cardiovascular risk assessment and diabetes. AB - Rigorously developed evidence based practice guidelines are necessary to promote clinical interventions that are consistent and most effective. Recommendations for treatment should be linked clearly to proven reduction in absolute risk, expressed as number needed to treat, rather than being based on the more commonly quoted and potentially misleading reduction in relative risk. Data published since 2003 has been incorporated in a 2009 revision of the New Zealand guideline for assessment and management of cardiovascular risk. Important changes are the decrease in optimal levels of LDL-C from 2 cm) require a formal oncologic resection. Adjuvant therapy is indicated only for metastatic disease, and admirable advances have been made in the realm of chemotherapy for reduction of disease and palliation of the symptoms of carcinoid syndrome. In this article, we discuss the nature of these interesting and uncommon tumors, clinical presentation, treatment options, and prognosis. PMID- 20011310 TI - Colorectal lymphoma. AB - Lymphoma involving the colon and rectum is rare and can occur either as primary colorectal lymphoma or as a manifestation of systemic disease. Most patients with primary colorectal lymphoma present with abdominal pain, and obstruction is unusual. Although lesions may be evident on radiographic studies, the findings are generally nonspecific. Historically, aggressive B-cell lymphomas have been the most common gastrointestinal lymphomas reported in the literature. However, recent reports suggest increased rates of mucosa-associated lymphoid tissue (MALT) lymphoma and mantle cell lymphoma (MCL) with increased surveillance.1 Most patients with colorectal lymphoma undergo surgery, but multiagent chemotherapy remains the mainstay of treatment. B-cell lymphomas tend to be most aggressive, with a median survival of about 2 years. PMID- 20011311 TI - Epidermoid carcinoma of the anal canal. AB - Anal cancers are rare tumors with only an expected 4000 new diagnoses in 2005. The majority of these are epidermoid or squamous cell cancers. Despite the rarity of this disease, great advances have been made in its understanding and treatment. The human papillomavirus (HPV), immunosuppression for solid organ transplantation, the human immunodeficiency virus (HIV), and sexual practices are changing the demographics of the disease from elderly women to young men who have sex with men and young women infected with HPV and HIV. The treatment of these malignancies was radically changed with Dr. Nigro's description in 1974 of the use of a nonoperative treatment strategy. Today, radiochemotherapy represents first-line therapy and the role of surgery has been largely relegated to that of salvage therapy for treatment failures of nonoperative management. PMID- 20011312 TI - Retrorectal tumors. AB - Primary neoplasms of the retrorectal (presacral) space are very rare. These lesions may be congenital or acquired, benign or malignant. They often arise with subtle clinical symptoms, or they may be found incidentally during evaluation for other conditions. Preoperative imaging can provide useful information for operative planning; however, it does not eliminate the need for surgery. Approaches for resection include posterior only and combined abdominoperineal, depending on the characteristics of the lesion. Successful treatment of these enigmatic lesions requires extensive knowledge of pelvic anatomy and expertise in pelvic surgery. PMID- 20011313 TI - Colorectal gastrointestinal stromal tumors: a brief review. AB - Gastrointestinal stromal tumors (GISTs) are rare lesions that constitute the majority of mesenchymal tumors in the gastrointestinal tract. Within the colon and rectum, they represent 0.1% of all cancers. They can present with a variety of symptoms but are often asymptomatic. Although many lesions may be benign, up to half of patients develop recurrent disease within a few years. Almost all GISTs contain a mutation in the c-kit tyrosine kinase that leads to its constitutive activation and results in cell proliferation. This discovery has led to the immunostaining of the c-kit antigen (CD117) to distinguish GISTs from other malignancies. Radiologic examinations can be helpful in initial diagnosis and staging. Surgery is the best treatment for cure, but recent advances have led to the use of imatinib mesylate, a tyrosine kinase inhibitor, to treat metastatic or unresectable disease, or both. There are currently many clinical trials available to help treat GISTs. PMID- 20011315 TI - Premalignant neoplasms and squamous cell carcinoma of the anal margin. AB - Premalignant and malignant lesions of the anal margin are rare. Understanding anal anatomy and performing a biopsy of any suspicious lesions are essential in avoiding a delay in diagnosis and appropriately treating these tumors. Wide local excision continues to remain the treatment of choice for many of these lesions. Combined multimodality treatment has come to play an important role in managing patient with more advanced or metastatic disease. PMID- 20011314 TI - Anal melanoma. AB - Anal melanoma is rare and aggressive malignancy. Patients commonly present with advanced, even metastatic disease. Unlike cutaneous melanoma, anal melanoma has no known risk factors. Surgical excision remains the cornerstone of therapy. There are no long-term survivors of stage II or III disease; therefore, early diagnosis and treatment remain crucial. There are no trials definitively proving abdominal perineal resection (APR) or wide local excision (WLE) to yield superior long-term survival. APR may offer a higher rate of local control, whereas WLE offers a much less morbid operation. Adjuvant chemotherapy, interferon, and radiation may offer some benefit. PMID- 20011316 TI - Hemangiomas of the pelvis. AB - Hemangiomas of the colon and rectum are rare but often misdiagnosed causes of hematochezia. They are characterized by the clinical triad of recurrent episodes of painless rectal bleeding, multiple ectopic phleboliths on plain radiographs, and cutaneous hemangiomas. The majority of these tumors involve the rectosigmoid region. Diffuse cavernous lesions are the most common histological type. Endoscopy is important to define the extent and number of lesions. Imaging by computed tomography or magnetic resonance imaging is used to define invasion into pelvic structures. Surgical resection with preservation of the sphincters is the recommended treatment. PMID- 20011318 TI - Reoperative surgery. PMID- 20011319 TI - Reoperation for small bowel obstruction--how critical is the timing? AB - The timing of reoperation for small bowel obstruction is a topic of significant debate. Any patient with evidence of strangulation should undergo urgent surgical intervention. However, predicting strangulation can be difficult. Because of this, previous authors have recommended everything from emergency operation for all patients presenting with small bowel obstruction to periods of observation that extend up to 14 days. Over the past century, the primary etiology of small bowel obstruction has shifted from hernias to postoperative adhesive disease, leading to a shift in the management paradigm. To manage small bowel obstruction successfully today, the clinician must distinguish the patient requiring urgent operation from those who benefit from nonoperative management. Furthermore, the clinician must be able to determine the appropriate length of time for conservative management. In this article we review the significant body of literature on this topic including the diagnostic workup and timing of potential operative intervention in the patient with small bowel obstruction. PMID- 20011320 TI - Surgical hemorrhage, damage control, and the abdominal compartment syndrome. AB - The morbidity and mortality associated with surgical hemorrhage are considerable, particularly when relaparotomy is necessary. This complication can usually be avoided with comprehensive preoperative patient evaluation and meticulous surgical technique. The damage control sequence is a useful surgical strategy when severe intraoperative coagulopathy or hemodynamic instability is present. Abdominal compartment syndrome is a potentially lethal phenomenon that can occur following laparotomy or large-volume fluid resuscitation, or both. Early recognition and intervention are critical to survival of the patient when this syndrome occurs. PMID- 20011322 TI - Reoperation for stoma-related complications. AB - Stoma complications are common. Most do not require reoperation, but when surgery is indicated, numerous options are available. Complications can arise early or late, and they can vary from benign to life-threatening. Meticulous preoperative planning is crucial in preventing stoma complications. Good communication with the patient is important in the decision-making process. PMID- 20011321 TI - Reoperative inflammatory bowel disease surgery. AB - With the advent of restorative proctocolectomy or ileal pouch-anal anastomosis (IPAA) for ulcerative colitis (UC), not only has there been potential for cure of UC but also patients have enjoyed marked improvements in bowel function, continence, and quality of life. However, IPAA can be complicated by postoperative small bowel obstruction, disease recurrence, and pouch failure secondary to pelvic sepsis, pouch dysfunction, mucosal inflammation, and neoplastic transformation. These may necessitate emergent or expeditious elective reoperation to salvage the pouch and preserve adequate function. Local, transanal, and transabdominal approaches to IPAA salvage are described, and their indications, outcomes, and the clinical parameters that affect the need for salvage are discussed. Pouch excision for failed salvage reoperation is reviewed as well. Relaparotomy is also frequently required for recurrent Crohn's disease (CD), especially given the nature of this as yet incurable illness. Risk factors for CD recurrence are examined, and the various surgical options and margins of resection are evaluated with a focus on bowel-sparing policy. Stricturoplasty, its outcomes, and its importance in recurrent disease are discussed, and segmental resection is compared with more extensive procedures such as total colectomy with ileorectal anastomosis. Lastly, laparoscopy is addressed with respect to its long-term outcomes, effect on surgical recurrence, and its application in the management of recurrent CD. PMID- 20011323 TI - Reoperation for anastomotic failure. AB - Anastomotic leak is a dreaded complication of colorectal surgery, with many potential causes. This complication carries with it a reported mortality ranging from 6 to 39%. Early diagnosis is key for the prevention of mortality. Here, we discuss the causes of an anastomotic leak and its signs and symptoms. Moreover, we explore the various modalities of diagnosis and treatment options. Most important, we discuss when a patient needs to be taken to the operating room and what procedure should be performed given various intraoperative findings. PMID- 20011324 TI - Reoperation following minimally invasive surgery: are the "rules" different? AB - This article discusses various indications for reoperation and how employing laparoscopy at primary operation might affect the incidence, presentation, and treatment of common complications. The abdomen is likely to be far less hostile after laparoscopic surgery than after laparotomy. Adhesions to the anterior abdominal wall are minimal or absent. As a result, relaparoscopy is a reasonable diagnostic and often successful treatment modality in patients suspected of having intra-abdominal complications following laparoscopic operation. Laparoscopic success in dealing with acute bowel obstruction after laparoscopic surgery is related to the paucity of adhesions and unique mechanisms of obstruction that are localized and amenable to minimal dissection. The same mechanisms are also responsible for the increased risk of bowel necrosis associated with bowel obstruction after laparoscopic surgery. Limited experience with successful laparoscopic management of bleeding and anastomotic leak has been reported with the caveat that if the bleeding or contamination is excessive, cannot be identified and controlled quickly, or is unresponsive to a reasonable and brief effort using laparoscopy, a prompt laparotomy is indicated. Based on the current literature, it is reasonable to conclude that laparoscopic approaches to primary Crohn's disease and relaparoscopy for recurrence are an appropriate (perhaps the most appropriate) management strategy. Also, laparoscopic restorative proctocolectomy and ileal pouch-anal anastomosis after laparoscopic subtotal colectomy is the preferred treatment for toxic ulcerative colitis. We conclude that laparoscopic reoperative surgery is feasible for the treatment of many complications following laparoscopic major abdominal surgery and bowel resection. PMID- 20011325 TI - Minimally Invasive Reoperation following Laparotomy. AB - The surgical indications for laparoscopic techniques continue to expand as experience is gained. This includes patients who have had a previous open abdominal operation and require surgical intervention. In addition to the generally accepted indications for laparoscopy, conditions such as reversal of colostomy, small bowel obstruction, and early reoperation for surgical complications may be managed using minimally invasive techniques. Specific considerations in this group of patients include peritoneal access and establishing domain, lysis of adhesions, and situations that should prompt conversion. PMID- 20011326 TI - Reoperation for recurrent colorectal cancer. AB - Recurrence of colorectal carcinoma represents a significant challenge. As the majority of recurrences involve more than just the anastomosis, surgical resection is ordinarily a major undertaking. Curative resection may require resection of other organs and structures, resulting in complex reconstructive procedures and substantial morbidity. In addition, carefully selected patients with distant metastases to sites such as the liver and lungs may also undergo potentially curative resection. Long-term survival following curative surgery for recurrence, however, ranges from only 15 to 40%. In addition to resection for curative intent, some patients may benefit from palliative procedures designed to relieve symptoms. Surgery alone is not usually sufficient therapy in these patients. Chemotherapy and radiation therapy play a vital adjunctive role in the management of recurrent disease. This article strives to review the risk factors and patterns of recurrence, selection of individuals for resection of recurrent disease, and outcomes of surgical procedures. PMID- 20011327 TI - Postoperative enterocutaneous fistula: when to reoperate and how to succeed. AB - An enterocutaneous fistula (ECF) is a potentially catastrophic postoperative complication. Although the morbidity and mortality associated with ECF have decreased over the past 50 years with modern medical and surgical care, the overall mortality is still surprisingly high, up to 39% in recent literature. It seems prudent, then, for every surgeon to have a thorough grasp of optimal treatment strategies for ECF to minimize their patients' mortality. Ultimately, the algorithm must begin with prevention. Once an ECF is diagnosed, the first step is to resuscitate and treat sepsis. The second is to control fistula output. The third step is to optimize the patient medically and nutritionally. The last step is definitive restoration of gastrointestinal continuity when necessary. Special mention is given in this article to exceptionally refractory fistulas such as those arising in the presence of inflammatory bowel disease and irradiated bowel. This plan gives a framework for the difficult task of successfully treating the postoperative ECF with a multidisciplinary approach. PMID- 20011328 TI - Reoperative surgery: what can we learn from a large randomized prospective trial? AB - Several recent large prospective trials and surveys of colon and rectal surgery patients have been conducted. Reviewing these studies can provide insight into current colorectal surgery practices. PMID- 20011329 TI - Can reoperative surgery be profitable? Maximizing reimbursement. AB - It is interesting to review the topic of reoperative surgery from the aspect of cost-effectiveness or, worse yet, profitability for either the institution or surgeon. The majority of the published material focuses on indications, risk factors, and short- and long-term outcomes. The context of the discussion is framed by the question "Can you make money doing reoperative surgery?" The short answer is that if this is all you do, probably not. Few, if any, publications assess the impact a redo procedure has on the factors that directly affect the fee schedule: surgeon time consumption (intraoperative and postoperative), stress (physical and psychological), and malpractice effects. Far more work needs to be done to understand the cost and resource consumption effects of major reoperative surgery. PMID- 20011333 TI - The cochrane collaboration: evidence-based medicine and common sense. PMID- 20011334 TI - Evaluation of anal incontinence: minimal approach, maximal effectiveness. AB - Anal incontinence is a symptom represented by the impaired ability to control the elimination of gas and stool, with an estimated incidence of 2.2 to 7.1% of the population. These numbers likely under-represent the true prevalence because physicians and patients are reluctant to discuss this problem. Evaluation of the patient with anal incontinence requires a fundamental knowledge of the etiologic factors. Careful history and physical examination is essential in every patient and can identify the cause of most cases of incontinence. Incontinence scoring systems are tools that provide objective data regarding the severity and quality of anal incontinence. Supplemental special tests for evaluating incontinence should be aimed at achieving three goals: (1) provide additional and confirmatory information regarding the diagnosis and cause of incontinence; (2) select appropriate treatment; and (3) predict treatment outcome. Numerous studies to evaluate anal incontinence exist; however, the most useful tests to achieve these goals are anal manometry, pudendal nerve terminal motor latency, and anal endosonography, because these studies can identify physiologic, neurologic, and anatomic abnormalities of the anorectum for which there may be effective treatments. PMID- 20011335 TI - Nonoperative management of fecal incontinence. AB - Fecal incontinence is a significant health problem that affects quality of life. Continence may often be improved with changes in diet with or without pharmacologic treatment to optimize the consistency of stool. Biofeedback is a multifaceted therapeutic approach that helps an individual improve his or her sensitivity to rectal distention and respond with appropriate voluntary sphincter control. One or more of these techniques can be used in conjunction with education and supportive counseling to optimize an individual's continence. PMID- 20011336 TI - Overlapping sphincteroplasty: is it the standard of care? AB - Anal sphincter injury secondary to obstetric trauma during vaginal delivery occurs in nearly one of every five women. Episiotomy, forceps delivery, and prolonged second stage of labor have all been shown to increase the risk of sphincter disruption. One third of these women will go on to have alterations in anal continence ranging from occasional incontinence to gas to severely debilitating incontinence to solid stool. Symptoms often arise many years after delivery, suggesting that factors such as nerve damage and progressive degeneration of muscle fibers contribute to incontinence. Surgical treatment of fecal incontinence secondary to sphincter injury has been varied and creative attempts have been made to find the repair with the greatest durability and fewest complications. Over the past few decades, overlapping sphincteroplasty emerged as such a repair with many reports of excellent short-term outcomes. Recently, however, published reports of long-term data reveal decreased function over time, causing many to question whether this repair truly is the best possible treatment. Several controversies have arisen. These include (1) optimum timing from injury to repair; (2) how best to perform the repair; (3) whether or not fecal diversion, either medical or surgical, is beneficial; (4) whether or not pudendal neuropathy predicts outcome; and finally, (5) if patient's age at the time of repair affects outcome. Randomized controlled trials are lacking, so any conclusions drawn from reviewing current literature must be evaluated with this in mind. Nonetheless, important information can be gleaned from the available literature and future studies designed with the hope of improving treatment for this life-altering condition. PMID- 20011337 TI - The current status of the Acticon Neosphincter. AB - Severe fecal incontinence is a very debilitating condition with a tremendous impact on an individual's life. In the past if a patient was not amenable to a tissue repair or failed a tissue repair, a colostomy was his or her only surgical option. New innovations have emerged giving patients more options to regain continence. The Acticon Neosphincter, which is an artificial bowel sphincter, is one of those newer options. The device is reserved for patients with severe fecal incontinence that is not amenable to lesser forms of therapy. Because it is an artificial device, the risk of infection is of particular concern. With experience, however, the infection rate has declined and the artificial bowel sphincter has become a good option for patients with severe fecal incontinence. The results are quite impressive with a significant number of patients obtaining complete continence. PMID- 20011338 TI - Sacral nerve stimulation for the treatment of fecal incontinence. AB - The use of sacral nerve stimulation as a treatment for fecal incontinence for intact but functionally deficient sphincter and pelvic floor musculature, as well as for some sphincter injuries, is an attractive concept that is currently undergoing clinical trials in the United States. Electrical stimulation of the peripheral nerve supply to the striated anal sphincter muscles at the level of the sacral spinal nerves exploits the accessibility of the most distal common location of the dual peripheral nerve supply to these muscles. While the mechanism of sacral nerve stimulation's salutary effect remains conjectural at present and is likely multifactorial, current experimental data point toward both an enhancement in striated muscular activity as well as neuromodulation of sacral reflexes that regulate rectal sensitivity and contractility. PMID- 20011339 TI - The Secca procedure for the treatment of fecal incontinence: definitive therapy or short-term solution. AB - The treatment of fecal incontinence by means of radiofrequency energy is based on the concept that collagen deposition and subsequent scarring may increase one's ability to recognize and retain stool and permit improved continence. The procedure is undertaken on an outpatient basis. Individuals may be considered candidates even if they have a potentially reparable defect since the technique does not limit one to the application of a subsequent procedure. Clearly, those for whom other treatment methods have failed and those who have no other reasonable option in the management of their fecal incontinence should be considered for this procedure. Preliminary results are quite encouraging, and the results of a prospective, sham-controlled, randomized clinical trial are awaited. PMID- 20011340 TI - Muscle transposition: does it still have a role? AB - Since the early 1900s, skeletal muscle transpositions have been employed for complicated cases of fecal incontinence to augment or replace the anal sphincter. Multiple techniques have evolved that vary with the type and configuration of muscle used in the reconstruction. Transposition of the gluteus maximus muscle was popular in the early stages of development but was replaced by techniques involving transposition of the gracilis muscle. Within the past 16 years, electrical stimulators have been applied to the transposed muscle flaps to create a dynamic reconstruction improving the efficacy of these neosphincters over their static counterparts. However, the stimulated versions are technically demanding with a high rate of morbidity secondary to complications of the multiple components and variations in technique. The stimulator used in this procedure has been removed from the US market, although it is still available in other countries. Currently in the United States, gracilis transposition is still employed in the absence of an electrical stimulator as an adjunct to the artificial bowel sphincter (Acticon Neosphincter, American Medical Systems, Minnetonka, MN), such as in cases of severe muscle loss and congenital atresia. In European countries, the stimulated graciloplasty continues to evolve, leading to expansion of its use in total anorectal reconstruction for anal atresia and after abdominoperineal resection. PMID- 20011341 TI - Complete pelvic floor repair in treating fecal incontinence. AB - Fecal incontinence is associated with 20 to 40% of the patients with pelvic floor prolapse. Successful management of fecal incontinence requires not only an understanding of anorectal function but also a thorough understanding of pelvic floor anatomy and how pelvic floor prolapse affects fecal continence. Imaging techniques have been instrumental in visualizing pelvic floor prolapse and have helped correlate surgical findings. Stabilization of the perineal body appears to be a key component to the success of pelvic floor repair and fecal continence, but the optimal repair is far from being established. PMID- 20011345 TI - Medical treatment of constipation. AB - Various agents are used for the medical management of chronic constipation, but few of these have been adequately studied. This article specifically examines the medical treatment of chronic constipation and the available data concerning bulk agents, lubricating agents, stimulants, and osmotic laxatives, used alone and in combination. Most experts consider dietary fiber or medicinal bulk agents to be the initial therapeutic option for the treatment of chronic constipation. If fiber is not successful or poorly tolerated, subsequent treatments may include saline osmotic laxatives, lactulose, 5-hydroxytryptamine4 (5-HT(4)) agonists (tegaserod), or stimulants such as senna or bisacodyl. Recent data also demonstrate both polyethylene glycol laxative and tegaserod to be safe and effective as initial therapy for chronic constipation. PMID- 20011344 TI - The evaluation of constipation. AB - Constipation is a major medical problem in the United States, affecting 2% to 28% of the population. Individual patients may have different conceptions of what constipation is, and the findings overlap with those in other functional gastrointestinal disorders. In 1999, an international panel of experts laid out specific criteria for the diagnosis of constipation known as the Rome II criteria. When patients present with complaints of constipation, a complete history and physical examination can elicit the cause of constipation. It is imperative to rule out a malignancy or other organic causes of the patient's symptoms prior to making the diagnosis of functional constipation. Many patients' symptoms can be relieved with lifestyle and dietary modification, both of which should be implemented before other potentially unnecessary tests are performed. Functional constipation is divided into two subtypes: slow transit constipation and obstructive defecation. Because many different terms are used interchangeably to describe these subtypes of constipation, physicians need to be comfortable with the language. Slow transit constipation is due to abnormal colonic motility. The diagnosis is made with the use of a colonic transit study. We continue to use a single-capsule technique as first described in the literature, but modifications of the capsule technique as well as scintigraphic techniques are validated and can be substituted in place of the capsule. Obstructive defecation is a much more complex problem, with etiologies ranging from rare diseases such as Hirschsprung's to physiologic abnormalities such as paradoxical puborectalis contraction. To fully evaluate the patient with obstructive defecation, anorectal manometry, defecography, and electromyography should be utilized. The different techniques available for each test are fully covered in this article. When evaluating each patient with constipation, it is important to keep in mind that the disease may be specific to one subtype or a combination of both subtypes. Because it is difficult to differentiate the subtypes from the patient's history, we feel it is imperative to evaluate patients fully for both slow transit and obstructive defecation prior to any surgical intervention. Furthermore, we have described many tests that need to be applied to one's population of patients on the basis of the capabilities and expertise the institution offers. PMID- 20011346 TI - Surgical management of constipation. AB - For the select small number of constipated patients who cannot be managed medically, surgical options should be considered. Increases in our knowledge of colorectal physiology and experience have fostered improvements in evaluation and surgical management of patients. Currently, patients with refractory colonic inertia are offered total abdominal colectomy and ileorectal anastomosis. With proper selection of patients, the results have been excellent for resolving the frequency and quality of bowel movements. However, symptoms such as bloating and abdominal pain, which may be related to irritable bowel syndrome rather than the colonic inertia, may persist. PMID- 20011348 TI - Acute intestinal pseudo-obstruction (Ogilvie's syndrome). AB - Acute colonic pseudo-obstruction (ACPO), also known as Ogilvie's syndrome, is a condition characterized by massive colonic distension in the absence of mechanical obstruction. Patients presenting with Ogilvie's syndrome have underlying medical and surgical conditions predisposing them to the syndrome. Ogilvie's syndrome can often be managed by conservative therapy. However, unrecognized and untreated, the continued distension associated with Ogilvie's syndrome can lead to perforation that is associated with a high mortality rate. In this article, the pathophysiology, epidemiology, and treatment options are reviewed. PMID- 20011347 TI - Treatment of obstructed defecation. AB - Obstructed defecation is a common problem that adversely affects the quality of life for many patients. Known causes of obstructed defecation include pelvic dyssynergy, rectocele, rectal intussusception, enterocele, pelvic organ prolapse, and overt rectal prolapse. Management of this condition requires an understanding of urinary, defecatory, and sexual function to achieve optimal outcomes. The goal of surgical treatment is to restore the various pelvic organs to their appropriate anatomic positions. However, there is a poor correlation between anatomic and functional results. It must be remembered that the pelvis contains many structures and that defects of pelvic support or function frequently affect other pelvic organs. Optimal outcomes can be achieved only by selecting appropriate treatment modalities that address all of the components of an individual patient's problem. PMID- 20011349 TI - Medical treatment of irritable bowel syndrome. AB - The identification, diagnosis, and treatment of irritable bowel syndrome (IBS) have a tremendous impact on the large number of patients with this syndrome and on physicians representing a wide variety of general and specialty practices. There are well-established diagnostic criteria and algorithms for the initial evaluation of patients presenting with the symptoms of IBS. The symptoms can be targeted for therapy with a variety of pharmaceutical and nonpharmaceutical agents. Therapy should be individualized for the patient, and the cornerstone for any effective treatment strategy should be a solid patient-physician relationship. Medications released in the past several years are based on the understanding of serotonin receptors in the gut and their role in the pathogenesis of IBS. PMID- 20011351 TI - Fecal impaction. AB - Fecal impaction is a common gastrointestinal problem and a potential source of major morbidity. Prompt identification and treatment minimize the risks of complications. Treatment options include manual extraction and proximal or distal washout. Following treatment, possible etiologies should be sought and preventive therapy instituted. PMID- 20011350 TI - Functional anorectal disorders. AB - Functional anorectal disorders include solitary rectal ulcer syndrome, rectocele, nonrelaxing puborectalis syndrome, and descending perineal syndrome. Patients usually present with "constipation," but the clinical picture of these disorders includes rectal pain and bleeding, digitalization, incomplete evacuation, and a feeling of obstruction. Diagnosis is difficult because many findings can be seen in normal patients as well. The diagnosis is made by using a combination of clinical picture, defecography, pathology, and occasionally anometry and pudendal terminal motor nerve latency. These disorders are generally treated medically with dietary changes and biofeedback. Surgical intervention is reserved for patients with intractable symptoms and has not been universally successful. PMID- 20011352 TI - Childhood constipation: evaluation and management. AB - Constipation is a common problem in children. It is also a long-term problem persisting for many months to years in children. Approximately 95% of childhood constipation is functional in nature without any obvious cause. Evaluation of a child with constipation requires a thorough history and physical examination. Hirschsprung's disease is an important cause of constipation arising in infancy and requires a thorough diagnostic evaluation and surgical treatment. Treatment of functional constipation in children requires a well-designed plan and a team approach involving the child, parents, and a health care provider. Treatment involves education of the family about constipation and encopresis, fecal disimpaction, and long-term maintenance therapy of laxatives and behavioral modification. Laxatives such as magnesium hydroxide, lactulose, and mineral oil have been used in children for a long time. A new laxative, polyethylene glycol 3350, has been used successfully in children with constipation and encopresis. Several novel therapeutic interventions have been tried for children presenting with intractable constipation, refractory to conventional treatment. PMID- 20011356 TI - Clostridium difficile colitis. AB - Clostridium difficile enterocolitis is endemic in most modern hospitals. The spectrum of clinical presentation varies from the asymptomatic carrier state to fulminant colitis with toxic megacolon and perforation. Highly toxigenic and lethal strains of C. difficile have emerged worldwide. Medical treatment consists of discontinuing the precipitating antibiotic, supportive measures and bowel rest, and antibiotic treatment with metronidazole or vancomycin. Surgical treatment may be necessary in cases of fulminant disease. Subtotal colectomy with end ileostomy is the operation of choice. PMID- 20011355 TI - Ischemic colitis. AB - Ischemic colitis is the most common form of gastrointestinal ischemia. Patients present with either occlusive or nonocclusive vascular disease, although the latter is more common. Many causes of nonocclusive disease have been identified, but the exact pathophysiology remains unclear. Most commonly, patients develop abdominal discomfort and bloody diarrhea. Diagnosis is confirmed with colonoscopy. Treatment is contingent on the severity of disease: mucosal/nongangrenous ischemia requires only supportive measures and medical management, whereas transmural/gangrenous ischemia may require prompt surgical intervention. Ischemic colitis can also become a chronic process with persistent segmental colitis or colonic stricturing. The patient's outcome depends on the severity of disease, prompt recognition, and appropriate treatment. PMID- 20011357 TI - Bacterial colitis. AB - Bacterial colitis results in an inflammatory-type diarrhea that is characterized by bloody, purulent, and mucoid stool. These diseases have been designated as bacterial hemorrhagic enterocolitis. Associated symptoms include fever, tenesmus, and severe abdominal pain. The pathologic changes range from superficial exudative enterocolitis to a transmural enterocolitis with ulceration. Common pathologic bacteria causing bacterial colitis include Campylobacter, Salmonella, Shigella, Escherichia, and Yersinia species. The primary source of transmission is fecal-oral spread and ingestion of contaminated food and water. Although detailed history and identification of specific risk factors assist in the diagnosis, definitive diagnosis requires bacterial identification. Therefore, the physician must be familiar with the disease pathophysiology, epidemiology, and specific diagnostic modalities for clinical diagnosis and management. Specific tests are used to detect enteric pathogens and include stool and rectal swab culture, histology, and identification of specific bacterial toxins. Although many of these bacterial colitis infections are self-limiting, antibiotics should be used for high-risk patients and patients with complicated disease. PMID- 20011358 TI - Viral and fungal infectious colitides. AB - Viral and fungal colitides are rare in the immunocompetent host but are most clinically significant in populations with compromised immune function. They may be associated with high mortality, particularly when treatment is delayed. It is important to be aware of these diseases when treating patients with colitis to allow early diagnosis and treatment, which will improve outcome. PMID- 20011359 TI - Amebiasis. AB - Amebiasis is an important cause of death from parasitic disease worldwide. The causative organism is Entamoeba histolytica, which has an infective cyst stage and a pathogenic and motile trophozoite stage. The clinical presentation can vary from an asymptomatic carrier state to fulminant colitis and colonic perforation. The majority of patients can be managed medically. However, a small percentage of patients require urgent exploration and resection with an associated high mortality rate. Early recognition and initiation of medical therapy including treatment of asymptomatic carriers are vital to preventing catastrophic outcomes. PMID- 20011360 TI - Parasitic colitides. AB - Parasitic infections are a major worldwide health problem, and they account for millions of infections and deaths each year. Most of the infections as well as the morbidity and mortality from these diseases occur in the developing world in rural regions. However, these diseases have become more common in Western countries and in big cities over the past 25 years. These changing disease patterns can be attributed to emigration from the third world to developed countries and migration of rural populations to the big cities in developing nations. These parasitic infections have protean manifestations and consequences. The medical problems range from chronic asymptomatic carrier to fulminant infections and even death. Several factors such as the host immune status, the infecting organism, and the availability of treatment all play key roles in the outcomes of parasitic colitides. The two major classes of parasites causing these infections are the helminthes (ascariasis, strongyloidiasis, enterobiasis, trichuriasis, and schistosomiasis) and the protozoa (Isospora, Cryptosporidium, Cyclospora, Trypanosoma cruzi, Giardia lamblia, and Balantidium coli). This article summarizes the salient features of each parasite with respect to epidemiology, transmission, pathogenesis, clinical features, diagnosis, and treatment. The vast majority of these infections have a self-limited clinical course or are easily treated with medical management, and surgery is rarely needed. PMID- 20011362 TI - Sexually transmitted proctitides. AB - Patients with sexually transmitted proctitides are increasingly presenting to doctors' offices. This may be secondary to increasing numbers of individuals participating in anal receptive intercourse and a rise in the incidence of sexually transmitted diseases. Although the sexually transmitted proctitides represent a small proportion of the overall number of cases of new proctitis, in certain populations the incidence of these diseases as causative agents is quite high, especially among men who have sex with men. Common causative agents include Neisseria gonorrhoeae, Chlamydia trachomatis, Treponema pallidum, and herpes simplex. Diagnosis may often be made on clinical grounds alone, and treatment requires antibiotics or antivirals. The clinician must remember to keep these diseases in mind while formulating a differential for the cause of proctitis. PMID- 20011361 TI - Noninfectious colitides: collagenous colitis, lymphocytic colitis, diversion colitis, and chemically induced colitis. AB - Approximately 10% of patients with chronic diarrhea carry a diagnosis of microscopic colitis. The endoscopic appearance of both collagenous colitis and lymphocytic colitis may be normal; however, biopsies confirm the diagnosis. Available treatments include antidiarrheals, bismuth salicylate, and budesonide. Although most patients with fecal diversion may have endoscopic evidence of colitis, a much smaller percentage of patients are symptomatic. Some cases of diversion colitis respond to treatment with short-chain fatty acid enemas; however, return of the fecal stream is the most successful therapy. A variety of oral, intravenous, and per rectum chemicals may cause colitis; symptoms usually abate when chemical exposure is discontinued. PMID- 20011366 TI - How to support military colon and rectal surgery. PMID- 20011363 TI - Radiation colitis and proctitis. AB - Radiation therapy is commonly utilized as a major component in the treatment of pelvic malignancy. Unfortunately, secondary toxicity to the lower gastrointestinal tract can occur. This most commonly affects the rectum, although injuries to the colon and small intestine are not uncommon. The presentation can be acute or chronic, and different mechanisms are responsible for each. Symptomatology is quite variable but can result in significant compromise for the patient. Numerous preventive and treatment strategies have been applied to this disease process. This article presents a summary of the current knowledge regarding radiation injury to the lower gastrointestinal tract with special emphasis on treatment options for radiation proctitis. PMID- 20011367 TI - Pursuing fellowship training in colorectal surgery: a candidate's perspective. AB - Colorectal surgery is a fulfilling profession that has become more difficult to match into. There are many factors that go into choosing a career in colorectal surgery. Having a mentor as a colorectal surgeon, performing research in colorectal surgery, and being involved with the American Society of Colon and Rectal Surgeons are all important aspects in providing exposure to young surgeons in training and guiding them toward the field of colorectal surgery. Once the decision has been made, the application and interview process can be daunting. This article reviews salient points about the process I took in choosing colorectal surgery and the steps I took in applying and interviewing for my fellowship. PMID- 20011368 TI - Career options in colon and rectal surgery. AB - As the field of colon and rectal surgery has expanded and diversified, so has the multitude of opportunities available to the prospective colon and rectal surgery graduate. Although this may initially appear exciting, these decisions may eventually become daunting and even paralyzing. However, prior to making a decision, candidates must first make an honest assessment of their goals, abilities, and priorities. This article briefly outlines some of these challenging choices and helps lay the groundwork for a successful decision process. PMID- 20011369 TI - Private practice for the colon and rectal surgeon. AB - Autonomy and independence as a surgeon represent the pinnacle of our training model, and private practice offers the trainee in colon and rectal surgery an opportunity to realize such goals as both a clinical surgeon and a business owner. Personalized care of patients and the immense gratification from providing such expert surgical care continue to be the ultimate reward for us as surgeons. However, private practice ultimately involves responsibilities of functioning as a small business owner. The health care environment in which we find ourselves provides great challenges to the viability and financial success of the private practitioner. Rising overhead expenses, malpractice, reduced reimbursement, and others factors confront the private practitioner as business owner. A career in private practice mandates acquisition of business acumen to preserve the privilege to practice our profession in this very challenging and changing economic environment. The opportunities for such a career vary considerably according to the scope of practice, hospital sophistication, geographic locale, and size of practice. PMID- 20011370 TI - Multispecialty clinic practice. AB - A multispecialty clinic practice is a common practice arrangement for colorectal surgeons. A practice of this type has a variety of features, both positive and negative. This article explores the location, practice patterns, lifestyles, compensation, and academic opportunities associated with a multispecialty clinic practice. This information can assist younger surgeons in choosing a practice opportunity and guide experienced surgeons' career progression. PMID- 20011371 TI - Military colon and rectal surgery practice. AB - Military medicine is a unique type of group practice. It can be very rewarding because of the quality of appreciative patients and fellow providers. The lifestyle has challenges, especially in time of war. This article describes many of the advantages and challenges of military medicine. PMID- 20011372 TI - Academic university practice: program selection and the interview process. AB - The decision to go into academic surgery, rather than private practice, is often multifactorial and includes the opportunity to participate in research, education, and patient care. The current job market for academic colon and rectal surgeons can be described as favorable and growing as there is a push for major academic institutions to obtain fellowship-trained colorectal surgeons. In selecting a job, one should be familiar with the department characteristics. This requires obtaining the answers to multiple questions and negotiation of institutional commitment. PMID- 20011374 TI - Research in academic colon and rectal surgery: keys to success. AB - In the current medical climate, there is increased pressure on colon and rectal surgeons to generate clinical revenue and decreased availability of time and resources to do research. For faculty who want to pursue research as part of an academic career, this article reviews some of the issues involved. The first half of the article discusses key issues in choosing the right job, such as institutional/departmental support and resources. The second half of the article discusses what is needed to make a research program successful, including funding mechanisms, mentors, collaborators, and the different requirements for basic science versus clinic research. PMID- 20011373 TI - Academic appointment and the process of promotion and tenure. AB - A critical component of a successful academic career is the understanding of institutional criteria and guidelines for academic appointment, promotion, and tenure. It is important to point out that these criteria and guidelines may vary from institution to institution; however, they are uniform for all clinical faculty within a single institution and do not differ from department to department. The purpose of this article is to provide the aspiring academic colon and rectal surgeon with a basic understanding of academic faculty appointments, promotion, and tenure. PMID- 20011375 TI - Education: a career of teaching in colon and rectal surgery. AB - The past 25 years have seen a revolution in the way surgery is learned, taught, and practiced. This revolution has increased the need for surgical educators to adapt surgical educational strategies to the modern practice environment. The purposes of this article are (1) to describe the impact of recent and upcoming changes in surgical education, (2) to explore the benefits of participating in surgical education activities both for academic surgeons and for surgeons in private practice, and (3) to review some of the avenues available to surgeons wishing to become involved or build a career in surgical education. PMID- 20011376 TI - Contracts. AB - Residents completing colon and rectal surgery training should take seriously the terms of contracts offered to them. Contracts should specify how and how much salary will be paid, what benefits the surgeon will receive, and what services are to be provided. However, contracts frequently fail to disclose what costs the surgeon will have to cover, what will lower that salary, what hours the surgeon must keep, and what time off the surgeon may expect. Signing a contract is easily done, but getting out of a contract to take a better job or move to another community may be difficult because of the wording in the contract. Surgeons need to have professional help to determine whether to sign a contract and what that contract should provide. An attorney experienced in representing physicians can be recommended by the medical association. PMID- 20011377 TI - Practice and hospital economics. AB - There has been a significant effort over the past 10 years to attempt to control the rate of increase in the cost of medical care. However, as is true of any economic system, there are multiple stakeholders involved and often competing motivations. The single largest source of medical inflation is the cost of pharmaceuticals; however, this topic is not directly discussed in this article Similarly, the cost of medical insurance products is not included as these issues cannot be directly addressed by physician behavior. The body of this discussion focuses on costs directly experienced by or potentially controlled by physicians. These areas include practice expense and margin and hospital direct costs and margin. It is essential for physicians to understand fully factors they can potentially control and areas they may be able to influence in this troubling era of cost containment. PMID- 20011378 TI - Retirement: a personal perspective of when, why, and how. AB - Retirement from colon and rectal surgery is usually a personal decision. The details of when, why, and how are individually specific and are shaped by our life experience, desires, personal and family commitments, and financial considerations. The reflections of two respected senior colon and rectal surgeons are presented to delineate some of these aspects. PMID- 20011381 TI - Hemorrhoids. AB - Hemorrhoids are normal vascular structures underlying the distal rectal mucosa and anoderm. Symptomatic hemorrhoidal tissues located above the dentate line are referred to as internal hemorrhoids and produce bleeding and prolapse. Thrombosis in external hemorrhoids results in painful swelling. Symptomatic internal hemorrhoids that fail bowel management programs may be amenable to in-office treatment with rubber band ligation or infrared coagulation. Internal hemorrhoids that fail to respond to these measures or complex internal and external hemorrhoidal disease may require a surgical hemorrhoidectomy, either open or closed. A stapled hemorrhoidopexy treats symptomatic internal hemorrhoids and should be employed with care and only after thorough training of the surgeon because of the risk of rare, severe complications. The choice of procedure should be based on the patient's symptoms, the extent of the hemorrhoidal disease, and the experience of the surgeon. PMID- 20011382 TI - Pilonidal disease. AB - Pilonidal disease is a common anorectal problem that typically affects young people. Numerous surgical procedures have been described, but treatment failure and disease recurrence are frequent, leading to considerable morbidity in these otherwise healthy patients. To manage this problem successfully, surgeons must consider the pathogenesis and presentation of the disease and weigh the advantages and disadvantages of any operation. Discussed in this article are the pathogenesis of pilonidal disease and basic treatment options for acute pilonidal abscesses, sinus tracts, and chronic or recurrent pilonidal disease. PMID- 20011383 TI - Rectovaginal fistulas: current surgical management. AB - Rectovaginal fistulas represent an often devastating condition in patients and a challenge for surgeons. Successful management of this condition must take into account a variety of variables including the etiology, size, and location of the fistula. Etiologies include obstetrical trauma, inflammatory bowel disease, malignant processes, and complications of radiation therapy and surgery. Repair options include local repairs, tissue transfer techniques, and abdominal operations. PMID- 20011384 TI - Perianal abscess/fistula disease. AB - Perirectal abscesses and fistulas represent the acute and chronic manifestations of the same disease process, an infected anal gland. They have beleaguered patients and physicians for millennia. A thorough understanding of the anatomy and pathophysiology of the disease process is critical for optimal diagnosis and management. Abscess management is fairly straightforward, with incision and drainage being the hallmark of therapy. Fistula management is much more complicated. It requires striking a balance between rates of healing and potential alteration of fecal continence. This, therefore, requires much more finesse. Many techniques are now available in the armamentarium of the surgeon who treats fistula-in-ano. Although no single technique is appropriate for all patients and all fistula types, appropriate selection of patients and choice of repair technique should yield higher success rates with lower associated morbidity. PMID- 20011386 TI - Fecal incontinence. AB - Fecal incontinence is a devastating problem for those afflicted. It can lead to embarrassment and social isolation. Treating the problem begins with a thorough history and physical examination. Further testing (such as anal physiology testing or anal endosonography) depends on the examination and treatment plan. Conservative medical therapies (such as dietary manipulation, medication to slow the stool, and biofeedback) are usually tried first. Surgery includes sphincter repair, SECCA, artificial bowel sphincter, and stoma. New therapies continue to evolve with the intention of further improving quality of life for these afflicted patients. PMID- 20011385 TI - Constipation and obstructed defecation. AB - Difficulties with bowel function are common and may be due to several causes including slow colonic transit and obstructed defecation. The anatomical and pathophysiological changes associated with these conditions are varying, often incompletely understood, and in many cases have limited treatment outcomes. Patients present with variable complaints and have previously tried a plethora of over-the-counter medications in an effort to relieve their symptoms. Physicians need an organized approach to manage these patients optimally. Improvements over the past few years in our understanding of the complex process of defecation, along with the increasing use of radiological and anorectal physiology studies, have led to improved treatment results. PMID- 20011387 TI - Rectal prolapse. AB - Despite the innovation of more than 100 surgical procedures for the treatment of complete rectal prolapse, no one procedure is best and applicable to all patients. Traditionally, procedures have been divided into abdominal and perineal approaches. The application of the laparoscopic approach to colon and rectal disease has allowed an additional less invasive method of therapy to treat rectal prolapse successfully. In comparison with conventional approaches, laparoscopy has achieved similar functional results and recurrence rates while reducing postoperative pain and hospital length of stay. PMID- 20011390 TI - Stomas and wound management. PMID- 20011388 TI - Anal fissure. AB - Anal fissure is a common problem, vexing to both patients and physicians. The historical mainstay of therapy has been some method of partial division of the internal anal sphincter with the serious potential complication of fecal incontinence. Nonsurgical treatment methods were therefore pursued, producing healing rates less than that seen after surgical therapy but none of the morbidity of surgery. This article summarizes accepted methods of modern medical and surgical therapy for anal fissure and offers a rationale for treatment type selection. PMID- 20011391 TI - Preoperative considerations and creation of normal ostomies. AB - Stomas provide fecal diversion in emergent and elective settings. Preoperative planning and counseling are extremely important to the creation of an acceptable and functional ostomy for the surgeon and patient. Proper site selection will help decrease the incidence of postoperative complications. Ileostomy, colostomy, and cecostomy indications and techniques are discussed. PMID- 20011392 TI - Technical tips for stoma creation in the challenging patient. AB - Stoma creation is a mental and technical exercise, often straightforward without any difficulty. However, creation of a protruding, tension free, well vascularized stoma in an obese individual with a thick abdominal wall and short, thickened mesentery can be a substantial challenge. Preoperative planning including stoma site marking, thoughtful consideration of all alternatives, and attention to technical detail will help create a stoma that will serve the ostomate well. The technical tips provided in this article should facilitate the process of selecting the appropriate intestinal segment, identifying the correct stoma site, and creating a functional stoma even in the most challenging situations. PMID- 20011393 TI - Early stomal complications. AB - The creation of intestinal stomas for diversion of enteric contents is an important component of the surgical management of several gastroenterologic disease processes. Despite the frequency with which these procedures are performed, complications of stoma creation remain common, despite extensive measures aimed at reducing them. Early postoperative complications (those seen less than one month postoperatively) can lead to significant cost, both financially and psychologically, and incur significant morbidity. Commonly seen early postoperative stomal complications include improper stoma site selection, vascular compromise, retraction, peristomal skin irritation, peristomal infection/abscess/fistula, acute parastomal herniation and bowel obstruction, and pure technical errors. The author reviews these early complications associated with stoma creation, discusses means of preventing them, and outlines the management strategy for such complications when they do occur. PMID- 20011395 TI - Peristomal dermatology. AB - Inflammatory or infectious conditions affecting the skin around an intestinal stoma are common and may be a source of considerable aggravation to patients. Recognition and prompt appropriate treatment of these conditions improves their quality of life. Peristomal pyoderma gangrenosum represents a most unusual peristomal skin condition that is difficult to treat; resolution often requires diagnosis and therapy of underlying inflammatory bowel disease. PMID- 20011394 TI - Late stomal complications. AB - Ostomy creation is a common surgical procedure performed by a variety of surgical specialties. Complications associated with stomas are frequent and run the gamut from technical, mechanical, physiologic, and psychologic. The impact of these complications ranges from simple inconvenience to life threatening. The majority of these complications may not occur for years following creation of the stoma. In this article, the author reviews many of the late complications associated with stomas and options regarding their management. PMID- 20011396 TI - Stomas and trauma. AB - The current dogma about the treatment of penetrating colon injuries is reviewed, both from the civilian and the military perspective. This discussion is still evolving, and the time-honored methods of diversion, including colostomy and ileostomy, are still appropriate for the most severe and devastating sorts of wounds, especially in the military context. For the vast majority of penetrating wounds, primary repair works well and should be practiced. For the few patients who have primary repair that fails and leaks, mortality rates are high. The art of surgery involves knowing when to divert and when to repair. PMID- 20011397 TI - Minimally invasive stomas. AB - Traditionally, stoma creation and end stoma reversal have been performed via a laparotomy incision. However, in many situations, stoma construction may be safely performed in a minimally invasive nature. This may include a trephine, laparoscopic, or combined approach. Furthermore, Hartmann's colostomy reversal, a procedure traditionally associated with substantial morbidity, may also be performed laparoscopically. The authors briefly review patient selection, preparation, and indications, and focus primarily on surgical techniques and results of minimally invasive stoma creation and Hartmann's reversal. PMID- 20011398 TI - Continent ileostomy: current status. AB - Continent ileostomy (Kock pouch) is an alternative to end ileostomy for patients who have undergone total proctocolectomy. The procedure reached its height of popularity in the late 1960s and early 1970s, but has been supplanted by restorative proctocolectomy, an operation that preserves the natural route of defecation. Continent ileostomy is still appropriate for selected patients with ulcerative colitis and familial polyposis who are not candidates for restorative proctocolectomy, for whom restorative proctocolectomy or end ileostomy have failed, and in a few other selected cases. Complication rates have decreased during the past three decades following technical improvements, but remain significant. In this review, the author summarizes the current indications for continent ileostomy, examines recent technical modifications, and discusses the management of complications. PMID- 20011399 TI - Abdominal wall modification for the difficult ostomy. AB - A select group of patients with major stomal problems may benefit from operative modification of the abdominal wall. Options may include a modified abdominoplasty (abdominal wall contouring), localized flaps, or liposuction. Although frequently successful, these techniques have the potential for significant morbidity. PMID- 20011402 TI - Volvulus, prolapse, intussusception, and functional disorders. PMID- 20011400 TI - Perineal wound complications after abdominoperineal resection. AB - Perineal wound complications following abdominoperineal resection (APR) is a common occurrence. Risk factors such as operative technique, preoperative radiation therapy, and indication for surgery (i.e., rectal cancer, anal cancer, or inflammatory bowel disease [IBD]) are strong predictors of these complications. Patient risk factors include diabetes, obesity, and smoking. Intraoperative perineal wound management has evolved from open wound packing to primary closure with closed suctioned transabdominal pelvic drains. Wide excision is used to gain local control in cancer patients, and coupled with the increased use of pelvic radiation therapy, we have experienced increased challenges with primary closure of the perineal wound. Tissue transfer techniques such as omental pedicle flaps, and vertical rectus abdominis and gracilis muscle or myocutaneous flaps are being used to reconstruct large perineal defects and decrease the incidence of perineal wound complications. Wound failure is frequently managed by wet to dry dressing changes, but can result in prolonged hospital stay, hospital readmission, home nursing wound care needs, and the expenditure of significant medical costs. Adjuvant therapies to conservative wound care have been suggested, but evidence is still lacking. The use of the vacuum-assisted closure device has shown promise in chronic soft tissue wounds; however, experience is lacking, and is likely due to the difficulty in application techniques. PMID- 20011403 TI - Volvulus: small bowel and colon. AB - Volvulus of the intestine is a surgical emergency. Volvulus of the small bowel is more common in children and is most often secondary to malrotation. Small bowel volvulus is an uncommon cause for small bowel obstruction in adults, and is more likely to be secondary to postoperative adhesive bands. Colonic volvulus is a rare cause of large bowel obstruction, but more common than small bowel volvulus. The sigmoid is most frequently involved, with redundant colon as the primary cause. Cecal volvulus most commonly is due to lack of fixation. Colonic volvulus has a specific radiographic appearance; however, small bowel volvulus is difficult to distinguish from other causes of small bowel obstruction by radiographic means. New surgical techniques with minimally invasive surgery are increasingly being applied to this old problem with good results in selected cases. PMID- 20011404 TI - Abdominal approaches for rectal prolapse. AB - The management of full-thickness rectal prolapse involves surgical intervention in the majority of cases. Many procedures have been described employing both perineal and abdominal approaches. Abdominal procedures result in more durable repair of the prolapse; however, the procedures require general anesthesia and are reserved for younger healthier patients. Laparoscopy has been utilized in the treatment of rectal prolapse since its introduction for colorectal procedures; recent studies have found equivalent long-term results and short-term outcomes. PMID- 20011405 TI - Perineal approaches to rectal prolapse. AB - The management of full-thickness rectal prolapse involves surgical intervention in the majority of cases. Many procedures have been described. Perineal procedures are generally reserved for patients with multiple comorbid conditions or those considered too elderly or frail to withstand an abdominal surgical approach. They also play an important role in the management of rectal mucosal prolapse. The techniques, advantages, and complications of perineal approaches to rectal prolapse in use today are the focus of this chapter. PMID- 20011406 TI - Intestinal intussusception. AB - Intussusception is defined as the invagination of one portion of the bowel into an immediately adjacent portion. Etiology, symptoms, diagnosis, and treatment are different in the pediatric and adult populations. In the pediatric population, most cases are idiopathic and result in the common scenario of ileocolic intussusception. Factors involved in causation include anatomic features of the developing gastrointestinal tract and infectious influences. In adults, the intussusceptum is typically the result of a mucosal, intramural, or extrinsic lead point that acts as a focal area of traction pulling the proximal portion of bowel into the peristalsing distal portion. The diagnosis and management in the pediatric population is relatively standardized with nonoperative reduction via air or contrast enemas attempted first. In the adult population, intussusception presents a preoperative diagnostic challenge; although surgical intervention is mandatory, intraoperative management remains controversial. PMID- 20011407 TI - Anorectal physiologic evaluation of constipation. AB - Chronic constipation is a widespread problem which affects approximately 63 million people in North America alone. Although the majority of patients respond to changes in dietary fiber and water intake, as well as the judicious use of laxatives, a small portion will not and as a result, will be evaluated by gastroenterologists and colorectal surgeons. These patients most frequently have slow transit constipation, pelvic floor dysfunction, or some combination of the two. The physiologic evaluation of these patients comprises colonic transit studies, anorectal manometry, cinedefecography, electromyography, and pudendal nerve terminal motor latency testing. In this article, the authors describe the physiologic basis for these studies and review the techniques used in diagnosing these conditions. PMID- 20011408 TI - Functional disorders: rectoanal intussusception. AB - Rectoanal intussusception (RI) is a telescoping of the rectal wall during defecation. RI is an easily recognizable physiologic phenomenon on defecography. The management, however, is much more controversial. Two predominant hypotheses exist regarding the etiology of RI: RI as a primary disorder, and RI as a secondary phenomenon. The diagnosis may be suspected based on clinical symptoms of obstructive defecation. Diagnostic modalities include defecography as the gold standard. Dynamic pelvic magnetic resonance imaging (DPMRI) and transperineal ultrasound are attractive alternatives to defecography; however, their sensitivity is poor in comparison to the gold standard at this time. Management strategies including conservative measures such as biofeedback and surgical procedures including mucosal proctectomy (Delorme), rectopexy, and stapled transanal rectal resection (STARR) procedures have varied degrees of efficacy. PMID- 20011409 TI - Functional disorders: rectocele. AB - The baseline prevalence of rectocele is not well defined as many women are asymptomatic and do not seek medical help. Gynecologists tend to perform posterior wall repairs more commonly than colorectal surgeons because they also address patients with vaginal symptoms in addition to those with defecatory dysfunction. Overall, surgical correction success rates for rectocele correction are quite high when using a vaginal approach. Vaginal dissection, as opposed to transrectal or transperineal approaches, results in better visualization and access to the endopelvic fascia and levator musculature, allowing for more firm anatomic correction. In addition, the maintenance of rectal mucosal integrity may reduce the risk of postoperative complications such as infection and fistula formation. With the rapidly growing popularity of synthetic and biologic implant kits in the field of pelvic reconstruction, outcomes data reporting is increasing and allowing surgeons to better understand the effect of various surgical techniques on vaginal, sexual, and defecatory symptoms. PMID- 20011410 TI - Paradoxical puborectalis contraction and increased perineal descent. AB - Paradoxical puborectalis contraction and increased perineal descent are two forms of functional constipation presenting as challenging diagnostic and treatment dilemmas to the clinician. In the evaluation of these disorders, the clinician should take special care to exclude anatomic disorders leading to constipation. Physical examination is supplemented by additional diagnostic modalities such as cinedefecography, electromyography, manometry, and pudendal nerve tefninal motor latency. Generally, these investigations should be used in combination with the two playing the more relied upon techniques. Treatment is typically conservative with biofeedback playing a principal role with favorable results when patient compliance is emphasized. When considering paradoxical puborectalis contraction, failure of biofeedback is usually augmented with botulinum toxin injection. Increased perineal descent is generally treated with biofeedback and perineal support maneuvers. Surgery has little or no role in these conditions. The patient who insists on surgical intervention for either of these two conditions should be offered a stoma. PMID- 20011411 TI - Slow transit constipation: a review of a colonic functional disorder. AB - Constipation is a common gastrointestinal complaint that can cause significant physical and psychosocial problems. It has been categorized as slow transit constipation, normal transit constipation, and obstructed defecation. Both the definition and pathophysiology of constipation are unclear, but attempts to describe each of the three types have been made. Slow transit constipation, a functional colonic disorder represents approximately 15 to 30% of constipated patients. The theorized etiologies are disorders of the autonomic and enteric nervous system and/or a dysfunctional neuroendocrine system. Slow transit constipation can be diagnosed with a complete history, physical exam, and a battery of specific diagnostic studies. Once the diagnosis is affirmed and medical management has failed, there are several treatment options. Biofeedback, sacral nerve stimulation, segmental colectomy, and subtotal colectomy with various anastomoses have all been used. Of those treatment options, a subtotal colectomy with ileorectal anastomosis is the most efficacious with the data to support its use. PMID- 20011413 TI - Radiologic and physiologic evaluation in colon and rectal surgery. PMID- 20011414 TI - Transrectal ultrasound, manometry, and pudendal nerve terminal latency studies in the evaluation of sphincter injuries. AB - Fecal incontinence may be due to postpartum anal sphincter injuries or neurological damage even in the absence of obvious perineal trauma. Anal physiologic testing with transrectal ultrasound, manometry, and pudendal nerve terminal latency studies help to identify those patients with anal sphincter injuries who might benefit from anal sphincter repair. In this article, the authors discuss the specific tests that are available and how to interpret them. PMID- 20011415 TI - Endorectal ultrasound: its role in the diagnosis and treatment of rectal cancer. AB - With development over the past 25 years of new surgical techniques and neoadjuvant therapy regimens for rectal cancer, physicians now have a range of treatment options that minimize morbidity and maximize the potential for cure. Accurate pretreatment staging is critical, ensuring adequate therapy and preventing overtreatment. Many options exist for staging primary rectal cancer. However, endorectal ultrasound (ERUS) remains the most attractive modality. It is an extension of the physical examination, and can be performed easily in the office. It is cost effective and is generally well tolerated by the patient, without need for general anesthesia. The authors discuss the data currently available on ERUS, including its accuracy and limitations, as well as the technical aspects of performing ERUS and interpreting the results. They also discuss new ultrasound technologies, which may improve rectal cancer staging in the future. PMID- 20011416 TI - Magnetic resonance imaging of rectal cancer. AB - Magnetic resonance imaging (MRI)is a useful modality for the evaluation of rectal cancer, providing superior anatomic/pathologic visualization when compared with endorectal ultrasound (EUS) and computed tomography (CT). Preoperative MRI is useful for tissue characterization and tumor staging, which determines the surgical approach and need for neoadjuvant/adjuvant therapy. Important prognostic factors include the circumferential resection margin (CRM), T and N stages, and extent of local invasion. Postoperative MRI to assess the extent of tumor recurrence enables early resection, which can greatly prolong survival. MRI criteria for local recurrence include T2 hyperintensity, early dynamic rim enhancement, and nodular morphology. Future research in MRI of rectal cancer is geared toward developing optimal imaging techniques including high-resolution MRI, whole-body scans, and parallel imaging; imaging of lymph nodes by MR lymphography; and response to therapy using diffusion/perfusion-weighted MR and functional imaging. PMID- 20011417 TI - Radiographic work-up and treatment of lower gastrointestinal bleeding. AB - Lower gastrointestinal hemorrhage is a common reason for hospital admission. Spontaneous cessation occurs in the majority of these patients; however, continued major bleeding is a difficult clinical problem. Emergency surgery, without prior knowledge of the bleeding site is associated with high morbidity and mortality rates. Accurate localization is therefore desirable. The authors present a review of current radiological imaging modalities and therapeutic options available to the clinician. They also provide a management algorithm to aid in the strategic management of this group of patients. PMID- 20011418 TI - Computed tomography enterography and magnetic resonance enterography: the future of small bowel imaging. AB - In the last 5 years, computed tomography enterography (CTE) and to a lesser extent magnetic resonance enterography (MRE) have supplanted the routine small bowel series and enteroclysis in the evaluation of many small bowel diseases, especially Crohn's disease. Both CTE and MRE use similar methods of bowel lumen opacification and distension and both have distinct advantages and disadvantages. Both have been most extensively studied in patients with Crohn's disease. What is certain is that these cross-sectional examinations have largely replaced the historic fluoroscopic examinations in the evaluation of the small bowel. PMID- 20011419 TI - Magnetic resonance imaging in the management of anal fistula and anorectal sepsis. AB - Complex perianal disease may be extremely debilitating for the patient with significant impingement on quality of life. The accurate identification of anatomical areas of involvement and subsequent appropriate management is crucial to achieving a successful outcome when treating anorectal sepsis and anal fistulae. Magnetic resonance imaging (MRI) has become a powerful tool in the evaluation of anal anatomy. In patients with complex disease MRI is an important adjunct in delineating disease location and extent, its relationship to sphincter muscles, and in planning management. MRI also plays an important role in evaluating the response to medical and surgical therapies. PMID- 20011420 TI - Computed tomography colonography (virtual colonoscopy): climax of a new era of validation and transition into community practice. AB - Colorectal cancer, which kills more than 50,000 patients every year in the United States and costs more than $6 billion in direct health costs, is a prime target for cancer prevention. Computed tomography colonography (CTC) has emerged as a minimally invasive, structural examination of the entire colon that can complement the current tools of cancer prevention and may improve patient compliance. Large trials have suggested a sensitivity of roughly 90% and specificity greater than 97% for CTC for patients with polyps >or= 10 mm. Bowel preparation by diet restriction, catharsis, and stool and fluid tagging are typically used. A prepless CTC protocol is an active area of research with a focus on improving patient compliance. Insurance coverage of CTC is a key factor affecting current dissemination and local and national coverage decisions are ongoing. CT examination of the abdomen allows visualization of extracolonic organs, where detection of additional disease must balance any unnecessary anxiety and testing. Estimates of CTC cost-effectiveness are generally favorable, but vary due to the high sensitivity of these models to costs, polyp sensitivity, compliance rates, and other parameters, which are difficult to accurately assess. Quality initiatives are being developed that will be key for implementation into community practice. PMID- 20011423 TI - Medical management of Crohn's disease. AB - The clinical course of Crohn's disease (CD) is characterized by unpredictable phases of disease activity and quiescence. The majority of CD patients experience mild to moderate disease or are in clinical remission over significant periods during the course of their disease. These patients can be treated conservatively with 5-aminosalicylates or budesonide depending on the disease location. Those patients with more severe forms of the disease who require corticosteroids should be treated more aggressively with early introduction of immunomodulator and/or biologic therapy, which may help to prevent the complications associated with CD. It has been suggested that therapies directed at mucosal healing may favorably modify the natural history of CD. As newer, more effective medications become available and new therapeutic approaches are introduced (top-down therapy), mucosal healing, and not solely clinical remission, may well become the preferred treatment objective. PMID- 20011424 TI - Perianal Crohn's disease. AB - Crohn's disease is commonly complicated by perianal manifestations. The surgeon plays a pivotal role in caring for these patients; a detailed history along with a thorough clinical exam provides the treating physician with invaluable information upon which to base further investigations and management decisions. Other than abscess drainage, medical management to control proximal disease often precedes any surgical attempt to cure the disease. Surgical interventions are indicated in selective patients, but are often complicated by poor wound healing and recurrences. A sizable percentage of these patients may need a proctectomy. PMID- 20011425 TI - Strictureplasty. AB - Strictureplasty in patients with Crohn's disease is an option in the colorectal surgeon's armamentarium for fibrostenotic obstructive disease. Common types include the Heineke-Mikulicz strictureplasty, Finney strictureplasty, and the side-to-side isoperistaltic strictureplasty. The procedure has potential for significant morbidity; therefore, it should be chosen for the patient carefully. Strictureplasty complements bowel resection in Crohn's disease; it is an excellent procedure to reduce the risk of developing short-bowel syndrome and its associated complications. PMID- 20011426 TI - Preoperative optimization of Crohn's disease. AB - Patients with Crohn's disease often present to the surgeon for operative intervention in poor overall condition. They may be taking multiple immunomodulators to attempt to manage their disease, may have significant weight loss and evidence of malnutrition, and 10 to 30% of the time will have intraabdominal sepsis in the form of an abscess or fistula. Preoperative optimization of these patients, when possible, may decrease morbidity and mortality, and may avoid formation of stomas for fecal diversion. Enhancing nutritional status and streamlining immunomodulator therapy prior to surgery may improve outcomes. Medical management of intraabdominal sepsis with percutaneous drainage of abdominal or pelvic abscesses may decrease postoperative septic complications, and may even avert the need for surgical intervention altogether. PMID- 20011427 TI - Colonic Crohn's disease. AB - Crohn's disease is an inflammatory disease of the gastrointestinal (GI) tract of uncertain etiology. It can affect any portion of the GI tract, involving the colon in approximately 60% of cases. Diagnosis can be unclear, but suspicion can be raised based upon clinical, endoscopic, and pathologic findings. Initial management is often medical, with surgery reserved for patients with colonic complications of Crohn's disease, such as hemorrhage, fulminant colitis, abscess or fistula, stricture, and malignant transformation. The operative choice and conduct depends upon the clinical presentation and intraoperative findings. The extent of resection is controversial, but segmental resection is appropriate in selected cases. PMID- 20011428 TI - Restorative operations for Crohn's disease. AB - Restoration of the continuity of the intestinal tract is one of the key concepts for maintaining the quality of life in patients with Crohn's disease. Restorative procedures have an important role in the scope of operative modalities for these patients. The authors review operative options aimed at fulfilling these goals including restorative partial small bowel resection; segmental, subtotal, and total colectomies; and ileal pouch anal anastomosis in patients with Crohn's disease. PMID- 20011430 TI - Laparoscopy in Crohn's disease. AB - Crohn's disease represents a challenging operative dilemma. The nature of the disease increases the technical complexity of operations, their morbidity, and the likelihood of multiple operations. In this setting, the advantages of laparoscopic surgery, including shorter hospital stays, less adhesion formation, fewer wound complications, and faster recovery of bowel function, are particularly beneficial to the patient. Patients with Crohn's disease requiring operations in the elective and semi-elective setting can all be approached initially laparoscopically. The surgeon's skill set should include extensive experience in advanced laparoscopic bowel surgery as well as open management of Crohn's disease and its complications. Strict adherence to the basic tenet of bowel preservation is imperative. The operations most commonly performed for Crohn's disease include diagnostic laparoscopy, stricturoplasty, small bowel resection, ileocolic resection, colectomy, repair of fistulae, and gastrojejunostomy for bypass of gastric or duodenal disease. Postoperative management includes resumption of steroids, typically without the need for "stress-dosing," bowel rest for a short period, and pain control, which is also less than that experienced with a laparotomy. PMID- 20011429 TI - Management of nonhealing perineal wounds. AB - The manifestations of perianal Crohn's disease vary from primary lesions such as skin tags and fissures, to diffuse septic destruction of tissue and sphincter muscle. These manifestations are often persistent and refractory to surgical treatment; however, a more disappointing scenario is when the treatment itself results in a chronic wound. The ideal approach for management involves basic surgical principles, careful patient selection, and realistic expectations. Choice of appropriate procedure, effective elimination of sepsis, thorough evaluation to rule out concomitant systemic disease, and appropriate use of fecal diversion are each an important principle. If proctectomy is necessary, several strategies such as intersphincteric dissection, avoidance of fecal contamination, and appropriate wound closure, are effective in diminishing the postoperative morbidity of an unhealed perineal wound. When an unhealed perineal wound develops in a patient with Crohn's disease, the initial management is conservative. When surgical treatment is necessary, success depends on careful patient selection, optimizing the patient's condition, elimination of sepsis, and choice of an effective technique for healing. PMID- 20011431 TI - Reoperative Crohn's surgery: tricks of the trade. AB - Crohn's disease is a chronic inflammatory disease of the intestinal tract that often results in the need for surgical intervention to treat complications of the disease. The transmural nature of the inflammation can lead to intestinal perforation, intraabdominal abscesses, intestinal strictures, and fistula development. Because there is no cure for Crohn's disease, many patients will require multiple operations during their lifetime. Index surgery and reoperative surgery in these patients is often complex and challenging. There are many preoperative planning and technical aspects of Crohn's surgery that can be helpful in achieving a successful clinical outcome. In this paper, we will review some of the important principles in operative and reoperative Crohn's surgery that can assist the surgeon when approaching these challenging cases. PMID- 20011433 TI - Polyps and polyposis coli. PMID- 20011434 TI - Polyp surveillance. AB - Surveillance after colonic polypectomy is important to detect and remove missed synchronous polyps and cancers and new metachronous polyps or cancers. The authors review methods of surveillance and the risk of recurrent adenomas and provide surveillance recommendations. PMID- 20011435 TI - Factors that increase risk of colon polyps. AB - Adenomatous polyps are common and factors that increase risk include race, gender, smoking, and obesity. This author summarizes the evidence supporting increased risk with these factors and describes how epidemiological data may be used to tailor screening programs. PMID- 20011436 TI - Colonic adenomatous polyposis syndromes: clinical management. AB - Colorectal cancer is one of the major causes of cancer deaths in both men and women. It is estimated that 5 to 10% of patients with colorectal cancer have an inherited germline mutation that predisposes them to cancer. Hereditary colorectal cancer syndromes can be divided into those associated with colonic polyposis - familial adenomatous polyposis (FAP), attenuated familial adenomatous polyposis (aFAP), and MYH associated polyposis (MAP), and those not associated with colonic polyposis - hereditary nonpolyposis colon cancer (HNPCC). The hereditary polyposes are usually easier to diagnose than HNPCC, but their higher penetrance and variable phenotype pose some difficult problems in management and surveillance. The timing and type of surgical intervention, the management of desmoid risk, the treatment of rectal or pouch neoplasia, and the management of duodenal neoplasia are all questions that must be addressed in patients with FAP or MAP. PMID- 20011437 TI - Extracolonic manifestations of hereditary colorectal cancer syndromes. AB - Familial colorectal adenocarcinoma (CRC) accounts for approximately 15 to 20% of CRC. Of these, hereditary nonpolyposis colorectal cancer (HNPCC) and familial adenomatous polyposis (FAP) represent the most common hereditary syndromes associated with CRC, followed by other less common diseases including juvenile polyposis (JP) and Peutz-Jeghers syndrome (PJS). Extracolonic manifestations are common in each of these syndromes having significant implications for surveillance and management in at-risk individuals. The authors review the most common and clinically relevant extracolonic manifestations for each of these syndromes focusing on incidence, presentation, genotype/phenotype correlations, and management (including surveillance) strategies. PMID- 20011438 TI - Management of serrated adenomas and hyperplastic polyps. AB - The benign serrated architecture of the hyperplastic polyp has now been recognized in morphologically similar lesions with potential for transformation to colorectal carcinoma: the sessile serrated adenoma (SSA), traditional serrated adenoma (TSA), and mixed polyp. These represent a group of serrated polyps with potential to evolve into colorectal cancer through a different molecular pathway than the traditional adenoma-carcinoma sequence, called the serrated pathway. Genetic characteristics involve a defect in apoptosis caused by BRAF and K-ras mutations that create distinct histologic characteristics of atypia in serrated architectural distortion of the crypts. An evidence-based algorithm for the clinical management of this polyp has yet to be determined. Current recommendations suggest these lesions be managed similar to conventional adenomas. The histology of serrated polyps is reviewed, as well as the common characteristics, and implications for treatment and surveillance. PMID- 20011439 TI - Polyps in children. AB - Children with polyps usually present with bleeding or pain. Most pediatric intestinal polyps are sporadic and are not associated with malignancy. Polyposis syndromes are also well described in children. Peutz-Jeghers syndrome is the most common hamartomatous polyposis condition. Although the polyps are not thought to be premalignant in most patients, there is an increased risk of other cancers. Familial adenomatous polyposis is also seen in childhood and is associated with a very high risk of malignant transformation as well as extracolonic adenomas and malignancy. The diagnosis and management of sporadic juvenile polyps, Peutz Jeghers syndrome, and familial adenomatous polyposis, as well as rarer conditions associated with intestinal polyps are reviewed in this article. PMID- 20011440 TI - Management of the malignant polyp. AB - In the United States, the prevalence of adenomatous polyps found during colonoscopic evaluation ranges from 25 to 41%, and of these, 2 to 5% contain invasive malignancy. The management of the malignant polyp continues to be challenging. Endoscopic resection by polypectomy has been shown to be sufficient for management of certain polyps containing cancer; however, it is important to keep in mind that polypectomy does not remove the lymph node drainage basin and may be an inadequate resection for some adenocarcinoma containing polyps that have specific histologic features. Depth of invasion has been shown to correlate with the risk of lymph node metastasis. Other unfavorable histologic features include lymphovascular invasion, poor differentiation, inability to assess margin (piecemeal resection), and positive resection margin (< 2 mm); these are important factors to consider in management. For these patients formal oncologic surgical resection is indicated. Traditional open or laparoscopic procedures are routinely used for colectomy in these patients. Following polypectomy or segmental colectomy, surveillance of these patients is critical, and can lead to excellent long-term outcomes. PMID- 20011441 TI - Local management of rectal neoplasia. AB - The treatment of rectal neoplasia, whether benign or malignant, challenges the surgeon. The challenge in treating rectal cancer is selecting the proper approach for the appropriate patient. In a small number of rectal cancer patients local excision may be the best approach. In an attempt to achieve two goals-cure of disease with a low rate of local failure and maintenance of function and quality of life-multiple approaches can be utilized. The key to obtaining a good outcome for any one patient is balancing the competing factors that impact on these goals. Any effective treatment aimed at controlling rectal cancer in the pelvis must take into account the disease in the bowel wall itself and the disease, or potential disease, in the mesorectum. The major downside of local excision techniques is the potential of leaving untreated disease in the mesorectum. Local management techniques avoid the potential morbidity, mortality, and functional consequences of a major abdominal radical resection and are thus quite effective in achieving the maintenance of function and quality of life goal. The issue for the transanal techniques is how they fare in achieving the first goal-cure of the cancer while keeping local recurrence rates to an absolute minimum. Without removing both the rectum and the mesorectum there is no completely accurate way to determine whether a rectal cancer has moved outside the bowel wall, so any decision on local management of a rectal neoplasm is a calculated risk. For benign neoplasia, the challenge is removing the lesion without having to resort to a major abdominal procedure. PMID- 20011442 TI - Polyps in the ileal pouch. AB - Total proctocolectomy and ileal pouch-anal anastomosis is the operation of choice for patients with familial adenomatous polyposis. With this operation comes the risk of developing ileal pouch polyps. Although rare, ileal pouch carcinomas may also occur within the pouch. Periodic endoscopic surveillance of the retained rectum and ileal pouch is recommended. Endoscopic polypectomy of medium and large polyps should be performed. Sulindac is effective in the reduction and often in the elimination of numerous smaller pouch polyps. Future studies are necessary to determine the role of sulindac and other chemotherapeutic agents in preventing the development of these polyps. PMID- 20011443 TI - Chemoprevention for colorectal neoplasia. AB - Colorectal cancer is a major cause of morbidity and mortality in industrialized societies and leads to significant treatment costs. Currently there are screening programs with fecal occult blood testing, radiographic, and endoscopic evaluation. Despite this, mortality from colorectal cancer has not improved dramatically. As such, attention has turned to finding agents to prevent carcinogenesis. The emerging field known as chemoprevention studies agents that target multiple steps in the development of adenomas and their subsequent transformation to colorectal carcinoma. There are multiple case control, cohort, and randomized controlled trials investigating the efficacy of fiber, calcium, vitamin D, folate, and nonsteroidal antiinflammatory drugs as chemopreventive agents against colorectal cancer. PMID- 20011444 TI - Work-up of the constipated patient. AB - Constipation is a common problem. Evaluation of patients should include a detailed history and clinical examination followed by radiologic and physiologic testing. The order of testing is dependent on patient symptoms and physician preference. The options are described along with their limitations. PMID- 20011447 TI - Immunologic and oncologic implications of laparoscopic surgery: what is the latest? AB - Laparoscopic surgery has been associated with many short-term benefits such as a shorter time to recovery, return of bowel function, less pain, and a decrease in wound infection rate. Several animal and human experiments have demonstrated an immunologic and oncologic benefit of minimally invasive surgery. Whether these results will translate into similar results in human settings is unclear. Although the first published prospective randomized clinical trial suggests better long-term outcomes for patients undergoing laparoscopic surgery, results from other ongoing randomized, controlled clinical trials are needed to verify this controversial result. PMID- 20011448 TI - Laparoscopy for rectal cancer: the need for randomized trials. AB - The adoption of laparoscopic proctectomy for rectal cancer has been relatively slow, primarily because of the technical difficulty of the procedure. The wide surgeon-to-surgeon variability in disease-free survival and local pelvic recurrence noted after open proctectomy is probably due to differences in surgical technique, and these differences are likely to be magnified when the additional challenge of laparoscopy is added to the procedure. At present, oncologic and functional outcomes data are limited. Although the adoption of laparoscopic techniques to perform curative proctectomy is likely to expand as technical challenges are overcome and experience and training improve, the results of prospective multicenter trials are necessary to ensure that the procedures provide an oncologic and functional outcome equivalent to that of conventional surgery. PMID- 20011449 TI - Hand-assisted laparoscopic colectomy: rational evolution for diverticulitis. AB - Laparoscopic technique has proved to be a revolutionary advance in the surgical treatment of disease. However, limits exist regarding its application to colorectal resection as evidenced by the higher conversion rate and longer learning curve seen with colectomy. Conversion remains a complex issue related to multiple factors. One of the factors, inflammatory disease such as diverticulitis, exposes limitations of laparoscopic technique, specifically the absence of tactile sensation and use of one's hand as a surgical instrument. Nonetheless, the clinical benefits of smaller incisions, decreased pain, decreased ileus, and reduced hospitalization and disability make laparoscopic colectomy a compelling surgical option for the treatment of diverticulitis. Hand assisted technique offers surgeons a practical and rational innovation for conventional laparoscopic colectomy and offers promise for improved feasibility and efficacy for the treatment of diverticulitis. PMID- 20011450 TI - Laparoscopy for inflammatory bowel disease: pushing the envelope. AB - Despite the slower learning curve of laparoscopic colectomy and the lack of prospective randomized trials, laparoscopic procedures have repeatedly demonstrated a shortened length of stay, reduction in postoperative ileus, and earlier resumption of diet. However, laparoscopy in inflammatory bowel disease has unique challenges that must be overcome. For the patient with uncomplicated terminal ileal Crohn's disease, there are definite reproducible advantages to a minimally invasive approach. As surgeons gain experience, more complex cases may be attempted laparoscopically with a low threshold to alternate the approach if difficulties are encountered. We will continue to "push the envelope" in patients with complex Crohn's disease to allow more to be done in complex cases. For the patient with Crohn's colitis and ulcerative colitis, the role of a minimally invasive approach is less well defined. In experienced hands, a laparoscopic total colectomy can be performed safely and offers the patient all the advantages seen with laparoscopic segmental resection. Outcomes are likely to improve with better training, techniques, and equipment. As the field of minimally invasive surgery continues to expand, what is being "pushed" today will be routine in the future. PMID- 20011451 TI - Laparoscopic colectomy in the elderly: when is too old? AB - To date, there is little literature regarding the impact of laparoscopic colectomy in the elderly population (i.e., patients older than 70 years) as the vast majority of studies regarding laparoscopic colectomy have evaluated younger patients (younger than 65 years). It is unknown whether elderly patients garner the same benefits from laparoscopic colectomy that younger patients have been shown to receive. As a result, there may be a reluctance to offer laparoscopy to elderly patients. The majority of the reports suggest that laparoscopic colectomy in the elderly is safe and provides the same benefits as laparoscopic colectomy in a younger population. Although an elderly population does not return to the work force, the benefits in the elderly population are related to a return to independence more often than after conventional surgery without an increase in hospital costs. Based on the current literature, one may never be too old to have a laparoscopic colectomy. PMID- 20011452 TI - Putting Electrospun Nanofibers to Work for Biomedical Research. AB - Electrospinning has been exploited for almost one century to process polymers and related materials into nanofibers with controllable compositions, diameters, porosities, and porous structures for a variety of applications. Owing to its high porosity and large surface area, a non-woven mat of electrospun nanofibers can serve as an ideal scaffold to mimic the extracellular matrix for cell attachment and nutrient transportation. The nanofiber itself can also be functionalized through encapsulation or attachment of bioactive species such as extracellular matrix proteins, enzymes, and growth factors. In addition, the nanofibers can be further assembled into a variety of arrays or architectures by manipulating their alignment, stacking, or folding. All these attributes make electrospinning a powerful tool for generating nanostructured materials for a range of biomedical applications that include controlled release, drug delivery, and tissue engineering. PMID- 20011453 TI - Paternal Work Characteristics and Father-Infant Interactions in Low-Income, Rural Families. AB - To examine the implications of paternal occupational conditions for the quality of father-infant interactions, home visits, including interviews and videotaped observations of father-infant interactions, were conducted with 446 fathers living in six low-income, nonmetropolitan counties in North Carolina and Pennsylvania. When a variety of individual and demographic characteristics were controlled for, a less supportive work environment was associated with lower levels of fathers' engaged and sensitive parenting. Significant interactions pointed to the importance of understanding combinations of risk factors. Experiencing high levels of workplace stressors, including low levels of self direction and high levels of care work, in the presence of other individual or demographic risk factors was associated with lower levels of father parenting quality. PMID- 20011454 TI - NORMAL FATTY ACID CONCENTRATIONS IN YOUNG CHILDREN WITH PHENYLKETONURIA (PKU). AB - The objective of this study was to determine if children with phenylketonuria (PKU) have lower fatty acid concentrations in total erythrocyte lipid due to the phenylalanine restricted diet therapy compared to healthy control subjects. Dietary intake and fatty acid concentrations in total erythrocyte lipid were measured in twenty-one subjects (99%) being stronger than HIV T. cruzi (approximately 90% for BaL and approximately 85% for VSV-G) infection. In MDM with established HIV-1 infection, T. cruzi significantly inhibited luciferate activity (p<0.01). By quantifying R-U5 and U5-gag transcripts by real time PCR, our study showed the expression of both transcripts significantly diminished in the presence of trypomastigotes (p<0.05). Thus, T. cruzi inhibits viral post-integration steps, early post-entry steps and entry into MDM. Trypomastigotes also caused a approximately 60-70% decrease of surface CCR5 expression on MDM. Multiplication of T. cruzi inside the MDM does not seem to be required for inhibiting HIV-1 replication since soluble factors secreted by trypomastigotes have shown similar effects. Moreover, the major parasite antigen cruzipain, which is secreted by the trypomastigote form, was able to inhibit viral production in MDM over 90% (p<0.01). CONCLUSIONS/SIGNIFICANCE: Our study showed that T. cruzi inhibits HIV-1 replication at several replication stages in macrophages, a major cell target for both pathogens. PMID- 20011522 TI - The use of knockout mice reveals a synergistic role of the Vav1 and Rasgrf2 gene deficiencies in lymphomagenesis and metastasis. AB - BACKGROUND: Vav1 and RasGRF2 are GDP/GTP exchange factors for Ras superfamily GTPases with roles in the development and/or effector functions of T-lymphocytes. METHODOLOGY/PRINCIPAL FINDINGS: Given that the phenotype of Vav1(-/-), Rasgrf2(-/ ) and Vav1(-/-);Rasgrf2(-/-) mice has been studied so far in young animals, we decided to explore the long-term consequences of the inactivation of those loci in the immune system. Unexpectedly, our studies revealed that the inactivation of the Vav1 proto-oncogene favors the formation of lymphoblastic lymphoma-like tumors in aging mice. Those tumors, that can be found either localized exclusively inside the thymus or widely disseminated in hematopoietic and non hematopoietic tissues, are composed of CD3(+) lymphoblasts that display heterogeneous combinations of CD4 and CD8 surface markers. Interestingly, the additional deletion of the Rasgrf2 gene induces a shortening in the latency period for the development of those tumors, an increase in the percentage of disseminated tumors outside the thymus and, as a result, higher mortality rates. CONCLUSIONS/SIGNIFICANCE: These data reveal unexpected negative roles for Vav1 and RasGRF2 in different stages of T-cell lymphoma progression. They also suggest that the inactivation of Vav1 function may represent an inadequate strategy to treat T-cell lymphomas, especially those associated with low levels of Rasgrf2 gene expression. PMID- 20011523 TI - Synchrotron x-ray visualisation of ice formation in insects during lethal and non lethal freezing. AB - Although the biochemical correlates of freeze tolerance in insects are becoming well-known, the process of ice formation in vivo is subject to speculation. We used synchrotron x-rays to directly visualise real-time ice formation at 3.3 Hz in intact insects. We observed freezing in diapausing 3(rd) instar larvae of Chymomyza amoena (Diptera: Drosophilidae), which survive freezing if it occurs above -14 degrees C, and non-diapausing 3(rd) instar larvae of C. amoena and Drosophila melanogaster (Diptera: Drosophilidae), neither of which survive freezing. Freezing was readily observed in all larvae, and on one occasion the gut was seen to freeze separately from the haemocoel. There were no apparent qualitative differences in ice formation between freeze tolerant and non-freeze tolerant larvae. The time to complete freezing was positively related to temperature of nucleation (supercooling point, SCP), and SCP declined with decreasing body size, although this relationship was less strong in diapausing C. amoena. Nucleation generally occurred at a contact point with the thermocouple or chamber wall in non-diapausing larvae, but at random in diapausing larvae, suggesting that the latter have some control over ice nucleation. There were no apparent differences between freeze tolerant and non-freeze tolerant larvae in tracheal displacement or distension of the body during freezing, although there was markedly more distension in D. melanogaster than in C. amoena regardless of diapause state. We conclude that although control of ice nucleation appears to be important in freeze tolerant individuals, the physical ice formation process itself does not differ among larvae that can and cannot survive freezing. This suggests that a focus on cellular and biochemical mechanisms is appropriate and may reveal the primary adaptations allowing freeze tolerance in insects. PMID- 20011524 TI - Patterned neuroprotection in the Inpp4a(wbl) mutant mouse cerebellum correlates with the expression of Eaat4. AB - The weeble mutant mouse has a frame shift mutation in inositol polyphosphate 4 phosphatase type I (Inpp4a). The phenotype is characterized by an early onset cerebellar ataxia and neurodegeneration, especially apparent in the Purkinje cells. Purkinje cell loss is a common pathological finding in many human and mouse ataxic disorders. Here we show that in the Inpp4a(wbl) mutant, Purkinje cells are lost in a specific temporal and spatial pattern. Loss occurs early in postnatal development; however, prior to the appearance of climbing fibers in the developing molecular layer, the mutant has a normal complement of Purkinje cells and they are properly positioned. Degeneration and reactive gliosis are present at postnatal day 5 and progress rapidly in a defined pattern of patches; however, Inpp4a is expressed uniformly across Purkinje cells. In late stage mutants, patches of surviving Purkinje cells appear remarkably normal with the exception that the climbing fibers have been excessively eliminated. Surviving Purkinje cells express Eaat4, a glutamate transporter that is differentially expressed in subsets of Purkinje cells during development and into adult stages. Prior to Purkinje cell loss, reactive gliosis and dendritic atrophy can be seen in Eaat4 negative stripes. Our data suggest that Purkinje cell loss in the Inpp4a(wbl) mutant is due to glutamate excitotoxicity initiated by the climbing fiber, and that Eaat4 may exert a protective effect. PMID- 20011525 TI - Nuclei of non-muscle cells bind centrosome proteins upon fusion with differentiating myoblasts. AB - BACKGROUND: In differentiating myoblasts, the microtubule network is reorganized from a centrosome-bound, radial array into parallel fibres, aligned along the long axis of the cell. Concomitantly, proteins of the centrosome relocalize from the pericentriolar material to the outer surface of the nucleus. The mechanisms that govern this relocalization are largely unknown. METHODOLOGY: In this study, we perform experiments in vitro and in cell culture indicating that microtubule nucleation at the centrosome is reduced during myoblast differentiation, while nucleation at the nuclear surface increases. We show in heterologous cell fusion experiments, between cultures of differentiating mouse myoblasts and human cells of non-muscular origin, that nuclei from non-muscle cells recruit centrosome proteins once fused with the differentiating myoblasts. This recruitment still occurs in the presence of cycloheximide and thus appears to be independent of new protein biosynthesis. CONCLUSIONS: Altogether, our data suggest that nuclei of undifferentiated cells have the dormant potential to bind centrosome proteins, and that this potential becomes activated during myoblast differentiation. PMID- 20011526 TI - N-cadherin negatively regulates osteoblast proliferation and survival by antagonizing Wnt, ERK and PI3K/Akt signalling. AB - BACKGROUND: Osteoblasts are bone forming cells that play an essential role in osteogenesis. The elucidation of the mechanisms that control osteoblast number is of major interest for the treatment of skeletal disorders characterized by abnormal bone formation. Canonical Wnt signalling plays an important role in the control of osteoblast proliferation, differentiation and survival. Recent studies indicate that the cell-cell adhesion molecule N-cadherin interacts with the Wnt co-receptors LRP5/6 to regulate osteoblast differentiation and bone accrual. The role of N-cadherin in the control of osteoblast proliferation and survival remains unknown. METHODS AND PRINCIPAL FINDINGS: Using murine MC3T3-E1 osteoblastic cells and N-cadherin transgenic mice, we demonstrate that N-cadherin overexpression inhibits cell proliferation in vitro and in vivo. The negative effect of N-cadherin on cell proliferation results from decreased Wnt, ERK and PI3K/Akt signalling and is restored by N-cadherin neutralizing antibody that antagonizes N-cadherin-LRP5 interaction. Inhibition of Wnt signalling using DKK1 or Sfrp1 abolishes the ability of N-cadherin blockade to restore ERK and PI3K signalling and cell proliferation, indicating that the altered cell growth in N cadherin overexpressing cells is in part secondary to alterations in Wnt signalling. Consistently, we found that N-cadherin overexpression inhibits the expression of Wnt3a ligand and its downstream targets c-myc and cyclin D1, an effect that is partially reversed by N-cadherin blockade. We also show that N cadherin overexpression decreases osteoblast survival in vitro and in vivo. This negative effect on cell survival results from inhibition of PI3K/Akt signalling and increased Bax/Bcl-2, a mechanism that is rescued by Wnt3a. CONCLUSION: The data show that N-cadherin negatively controls osteoblast proliferation and survival via inhibition of autocrine/paracrine Wnt3a ligand expression and attenuation of Wnt, ERK and PI3K/Akt signalling, which provides novel mechanisms by which N-cadherin regulates osteoblast number. PMID- 20011527 TI - Molecular characterization of pneumococcal isolates from pets and laboratory animals. AB - BACKGROUND: Between 1986 and 2008 Streptococcus pneumoniae was isolated from 41 pets/zoo animals (guinea pigs (n = 17), cats (n = 12), horses (n = 4), dogs (n = 3), dolphins (n = 2), rat (n = 2), gorilla (n = 1)) treated in medical veterinary laboratories and zoos, and 44 laboratory animals (mastomys (multimammate mice; n = 32), mice (n = 6), rats (n = 4), guinea pigs (n = 2)) during routine health monitoring in an animal facility. S. pneumoniae was isolated from nose, lung and respiratory tract, eye, ear and other sites. METHODOLOGY/PRINCIPAL FINDINGS: Carriage of the same isolate of S. pneumoniae over a period of up to 22 weeks was shown for four mastomys. Forty-one animals showed disease symptoms. Pneumococcal isolates were characterized by optochin sensitivity, bile solubility, DNA hybridization, pneumolysin PCR, serotyping and multilocus sequence typing. Eighteen of the 32 mastomys isolates (56%) were optochin resistant, all other isolates were optochin susceptible. All mastomys isolates were serotype 14, all guinea pig isolates serotype 19F, all horse isolates serotype 3. Rats had serotypes 14 or 19A, mice 33A or 33F. Dolphins had serotype 23F, the gorilla serotype 14. Cats and dogs had many different serotypes. Four isolates were resistant to macrolides, three isolates also to clindamycin and tetracycline. Mastomys isolates were sequence type (ST) 15 (serotype 14), an ST/serotype combination commonly found in human isolates. Cats, dogs, pet rats, gorilla and dolphins showed various human ST/serotype combinations. Lab rats and lab mice showed single locus variants (SLV) of human STs, in human ST/serotype combinations. All guinea pig isolates showed the same completely new combination of known alleles. The horse isolates showed an unknown allele combination and three new alleles. CONCLUSIONS/SIGNIFICANCE: The isolates found in mastomys, mice, rats, cats, dogs, gorilla and dolphins are most likely identical to human pneumococcal isolates. Isolates from guinea pigs and horses appear to be specialized clones for these animals. Our data redraw attention to the fact that pneumococci are not strictly human pathogens. Pet animals that live in close contact to humans, especially children, can be infected by human isolates and also carriage of even resistant isolates is a realistic possibility. PMID- 20011528 TI - Stat1 nuclear translocation by nucleolin upon monocyte differentiation. AB - BACKGROUND: Members of the signal transducer and activator of transcription (Stat) family of transcription factors traverse the nuclear membrane through a specialized structure, called the nuclear pore complex (NPC), which represents a selective filter for the import of proteins. Karyophilic molecules can bind directly to a subset of proteins of the NPC, collectively called nucleoporins. Alternatively, the transport is mediated via a carrier molecule belonging to the importin/karyopherin superfamily, which transmits the import into the nucleus through the NPC. METHODOLOGY/PRINCIPAL FINDINGS: In this study, we provide evidence for an alternative Stat1 nuclear import mechanism, which is mediated by the shuttle protein nucleolin. We observed Stat1-nucleolin association, nuclear translocation and specific binding to the regulatory DNA element GAS. Using expression of nucleolin transgenes, we found that the nuclear localization signal (NLS) of nucleolin is responsible for Stat1 nuclear translocation. We show that this mechanism is utilized upon differentiation of myeloid cells and is specific for the differentiation step from monocytes to macrophages. CONCLUSIONS/SIGNIFICANCE: Our data add the nucleolin-Stat1 complex as a novel functional partner for the cell differentiation program, which is uniquely poised to regulate the transcription machinery via Stat1 and nuclear metabolism via nucleolin. PMID- 20011529 TI - Residual antimalarials in malaria patients from Tanzania--implications on drug efficacy assessment and spread of parasite resistance. AB - BACKGROUND: Repeated antimalarial treatment for febrile episodes and self treatment are common in malaria-endemic areas. The intake of antimalarials prior to participating in an in vivo study may alter treatment outcome and affect the interpretation of both efficacy and safety outcomes. We report the findings from baseline plasma sampling of malaria patients prior to inclusion into an in vivo study in Tanzania and discuss the implications of residual concentrations of antimalarials in this setting. METHODS AND FINDINGS: In an in vivo study conducted in a rural area of Tanzania in 2008, baseline plasma samples from patients reporting no antimalarial intake within the last 28 days were screened for the presence of 14 antimalarials (parent drugs or metabolites) using liquid chromatography-tandem mass spectrometry. Among the 148 patients enrolled, 110 (74.3%) had at least one antimalarial in their plasma: 80 (54.1%) had lumefantrine above the lower limit of calibration (LLC = 4 ng/mL), 7 (4.7%) desbutyl-lumefantrine (4 ng/mL), 77 (52.0%) sulfadoxine (0.5 ng/mL), 15 (10.1%) pyrimethamine (0.5 ng/mL), 16 (10.8%) quinine (2.5 ng/mL) and none chloroquine (2.5 ng/mL). CONCLUSIONS: The proportion of patients with detectable antimalarial drug levels prior to enrollment into the study is worrying. Indeed artemether lumefantrine was supposed to be available only at government health facilities. Although sulfadoxine-pyrimethamine is only recommended for intermittent preventive treatment in pregnancy (IPTp), it was still widely used in public and private health facilities and sold in drug shops. Self-reporting of previous drug intake is unreliable and thus screening for the presence of antimalarial drug levels should be considered in future in vivo studies to allow for accurate assessment of treatment outcome. Furthermore, persisting sub-therapeutic drug levels of antimalarials in a population could promote the spread of drug resistance. The knowledge on drug pressure in a given population is important to monitor standard treatment policy implementation. PMID- 20011530 TI - Whole genome duplications and a 'function' for junk DNA? Facts and hypotheses. AB - BACKGROUND: The lack of correlation between genome size and organismal complexity is understood in terms of the massive presence of repetitive and non-coding DNA. This non-coding subgenome has long been called "junk" DNA. However, it might have important functions. Generation of junk DNA depends on proliferation of selfish DNA elements and on local or global DNA duplication followed by genic non functionalization. METHODOLOGY/PRINCIPAL FINDINGS: Evidence from genomic analyses and experimental data indicates that Whole Genome Duplications (WGD) are often followed by a return to the diploid state, through DNA deletions and intra/interchromosomal rearrangements. We use simple theoretical models and simulations to explore how a WGD accompanied by sequence deletions might affect the dosage balance often required among several gene products involved in regulatory processes. We find that potential genomic deletions leading to changes in nuclear and cell volume might potentially perturb gene dosage balance. CONCLUSIONS/SIGNIFICANCE: The potentially negative impact of DNA deletions can be buffered if deleted genic DNA is, at least temporarily, replaced by repetitive DNA so that the nuclear/cell volume remains compatible with normal living. Thus, we speculate that retention of non-functionalized non-coding DNA, and replacement of deleted DNA through proliferation of selfish elements, might help avoid dosage imbalances in cycles of polyploidization and diploidization, which are particularly frequent in plants. PMID- 20011531 TI - Tropical mosquito assemblages demonstrate 'textbook' annual cycles. AB - BACKGROUND: Annual biological rhythms are often depicted as predictably cyclic, but quantitative evaluations are few and rarely both cyclic and constant among years. In the monsoon tropics, the intense seasonality of rainfall frequently drives fluctuations in the populations of short-lived aquatic organisms. However, it is unclear how predictably assemblage composition will fluctuate because the intensity, onset and cessation of the wet season varies greatly among years. METHODOLOGY/PRINCIPAL FINDINGS: Adult mosquitoes were sampled using EVS suction traps baited with carbon dioxide around swamplands adjacent to the city of Darwin in northern Australia. Eleven sites were sampled weekly for five years, and one site weekly for 24 years, the sample of c. 1.4 million mosquitoes yielding 63 species. Mosquito abundance, species richness and diversity fluctuated seasonally, species richness being highly predictable. Ordination of assemblage composition demonstrated striking annual cycles that varied little from year to year. The mosquito assemblage was temporally structured by a succession of species peaks in abundance. CONCLUSION/SIGNIFICANCE: Ordination provided strong visual representation of annual rhythms in assemblage composition and the means to evaluate variability among years. Because most mosquitoes breed in shallow freshwater which fluctuates with rainfall, we did not anticipate such repeatability; we conclude that mosquito assemblage composition appears adapted to predictable elements of the rainfall. PMID- 20011532 TI - Tension at the surface: which phase is more important, liquid or vapor? AB - Tension at the surface is a most fundamental physicochemical property of a liquid surface. The concept of surface tension has widespread implications in numerous natural, engineering and biomedical processes. Research to date has been largely focused on the liquid side; little attention has been paid to the vapor--the other side of the surface, despite over 100 years of study. However, the question remains as to whether the vapor plays any role, and to what extent it affects the surface tension of the liquid. Here we show a systematic study of the effect of vapor on the surface tension and in particular, a surprising observation that the vapor, not the liquid, plays a dominant role in determining the surface tension of a range of common volatile organic solutions. This is in stark contrast to results of common surfactants where the concentration in the liquid plays the major role. We further confirmed our results with a modified adsorption isotherm and molecular dynamics simulations, where highly structured, hydrogen bonded networks, and in particular a solute depletion layer just beneath the Gibbs dividing surface, were revealed. PMID- 20011533 TI - Resting network plasticity following brain injury. AB - The purpose of this study was to examine neural network properties at separate time-points during recovery from traumatic brain injury (TBI) using graph theory. Whole-brain analyses of the topological properties of the fMRI signal were conducted in 6 participants at 3 months and 6 months following severe TBI. Results revealed alterations of network properties including a change in the degree distribution, reduced overall strength in connectivity, and increased "small-worldness" from 3 months to 6 months post injury. The findings here indicate that, during recovery from injury, the strength but not the number of network connections diminishes, so that over the course of recovery, the network begins to approximate what is observed in healthy adults. These are the first data examining functional connectivity in a disrupted neural system during recovery. PMID- 20011534 TI - Bi-directional SIFT predicts a subset of activating mutations. AB - Advancements in sequencing technologies have empowered recent efforts to identify polymorphisms and mutations on a global scale. The large number of variations and mutations found in these projects requires high-throughput tools to identify those that are most likely to have an impact on function. Numerous computational tools exist for predicting which mutations are likely to be functional, but none that specifically attempt to identify mutations that result in hyperactivation or gain-of-function. Here we present a modified version of the SIFT (Sorting Intolerant from Tolerant) algorithm that utilizes protein sequence alignments with homologous sequences to identify functional mutations based on evolutionary fitness. We show that this bi-directional SIFT (B-SIFT) is capable of identifying experimentally verified activating mutants from multiple datasets. B-SIFT analysis of large-scale cancer genotyping data identified potential activating mutations, some of which we have provided detailed structural evidence to support. B-SIFT could prove to be a valuable tool for efforts in protein engineering as well as in identification of functional mutations in cancer. PMID- 20011535 TI - Caveolae act as membrane reserves which limit mechanosensitive I(Cl,swell) channel activation during swelling in the rat ventricular myocyte. AB - BACKGROUND: Many ion channels are preferentially located in caveolae where compartmentalisation/scaffolding with signal transduction components regulates their activity. Channels that are mechanosensitive may be additionally dependent on caveolar control of the mechanical state of the membrane. Here we test which mechanism underlies caveolar-regulation of the mechanosensitive I(Cl,swell) channel in the adult cardiac myocyte. METHODOLOGY/PRINCIPAL FINDINGS: Rat ventricular myocytes were exposed to solution of 0.02 tonicity (T; until lysis), 0.64T for 10-15 min (swelling), and/or methyl-beta-cyclodextrin (MBCD; to disrupt caveolae). MBCD and 0.64T swelling reduced the number of caveolae visualised by electron microscopy by 75 and 50% respectively. MBCD stimulated translocation of caveolin 3 from caveolae-enriched buoyant membrane fractions, but both caveolin 1 and 3 remained in buoyant fractions after swelling. I(Cl,swell) inhibition in control cells decreased time to half-maximal volume (t(0.5,vol); 0.64T), consistent with a role for I(Cl,swell) in volume regulation. MBCD-treated cells showed reduced time to lysis (0.02T) and t(0.5,vol) (0.64T) compared with controls. The negative inotropic response to swelling (an index of I(Cl,swell) activation) was enhanced by MBCD. CONCLUSIONS/SIGNIFICANCE: These data show that disrupting caveolae removes essential membrane reserves, which speeds swelling in hyposmotic conditions, and thereby promotes activation of I(Cl,swell). They illustrate a general principle whereby caveolae as a membrane reserve limit increases in membrane tension during stretch/swelling thereby restricting mechanosensitive channel activation. PMID- 20011536 TI - Human erythrocytes selectively bind and enrich infectious HIV-1 virions. AB - Although CD4(+) cells represent the major target for HIV infection in blood, claims of complement-independent binding of HIV-1 to erythrocytes and the possible role of Duffy blood group antigen, have generated controversy. To examine the question of binding to erythrocytes, HIV-1 was incubated in vitro with erythrocytes from 30 healthy leukapheresis donors, and binding was determined by p24 analysis and adsorption of HIV-1 with reduction of infectivity for CD4(+) target cells. All of the cells, regardless of blood group type, bound HIV-1 p24. A typical preparation of erythrocytes bound <2.4% of the added p24, but erythrocytes selectively removed essentially all of the viral infectivity as determined by decreased infection of CD4(+) target cells; however, cell associated HIV-1 was approximately 100-fold more efficient, via trans infection, than unadsorbed virus for infection of CD4(+) cells. All of the bound HIV-1 p24 was released by treatment of the cells with EDTA, and binding was optimized by adding Ca(2+) and Mg(2+) during the washing of erythrocytes containing bound HIV 1. Although the small number of contaminating leukocytes in the erythrocyte preparation also bound HIV-1 p24, there was no significant binding to CD4, and it thus appears that the binding occurred on leukocytes at non-CD4 sites. Furthermore, binding occurred to erythrocyte ghosts from which contaminating leukocytes had been previously removed. The results demonstrate that erythrocytes incubated in vitro with HIV-1 differentially adsorb all of the infectious HIV-1 virions (as opposed to non-infectious or degraded virions) in the absence of complement and independent of blood group, and binding is dependent on divalent cations. By analogy with HIV-1 bound to DC-SIGN on dendritic cells, erythrocyte bound HIV-1 might comprise an important surface reservoir for trans infection of permissive cells. PMID- 20011537 TI - RNA-interference knockdown of Drosophila pigment dispersing factor in neuronal subsets: the anatomical basis of a neuropeptide's circadian functions. AB - BACKGROUND: In animals, neuropeptide signaling is an important component of circadian timekeeping. The neuropeptide pigment dispersing factor (PDF) is required for several aspects of circadian activity rhythms in Drosophila. METHODOLOGY/PRINCIPAL FINDINGS: Here we investigate the anatomical basis for PDF's various circadian functions by targeted PDF RNA-interference in specific classes of Drosophila neuron. We demonstrate that PDF is required in the ventro lateral neurons (vLNs) of the central brain and not in the abdominal ganglion for normal activity rhythms. Differential knockdown of PDF in the large or small vLNs indicates that PDF from the small vLNs is likely responsible for the maintenance of free-running activity rhythms and that PDF is not required in the large vLNs for normal behavior. PDF's role in setting the period of free-running activity rhythms and the proper timing of evening activity under light:dark cycles emanates from both subtypes of vLN, since PDF in either class of vLN was sufficient for these aspects of behavior. CONCLUSIONS/SIGNIFICANCE: These results reveal the neuroanatomical basis PDF's various circadian functions and refine our understanding of the clock neuron circuitry of Drosophila. PMID- 20011538 TI - Ethacrynic acid exhibits selective toxicity to chronic lymphocytic leukemia cells by inhibition of the Wnt/beta-catenin pathway. AB - BACKGROUND: Aberrant activation of Wnt/beta-catenin signaling promotes the development of several cancers. It has been demonstrated that the Wnt signaling pathway is activated in chronic lymphocytic leukemia (CLL) cells, and that uncontrolled Wnt/beta-catenin signaling may contribute to the defect in apoptosis that characterizes this malignancy. Thus, the Wnt signaling pathway is an attractive candidate for developing targeted therapies for CLL. METHODOLOGY/PRINCIPAL FINDINGS: The diuretic agent ethacrynic acid (EA) was identified as a Wnt inhibitor using a cell-based Wnt reporter assay. In vitro assays further confirmed the inhibitory effect of EA on Wnt/beta-catenin signaling. Cell viability assays showed that EA selectively induced cell death in primary CLL cells. Exposure of CLL cells to EA decreased the expression of Wnt/beta-catenin target genes, including LEF-1, cyclin D1 and fibronectin. Immune co-precipitation experiments demonstrated that EA could directly bind to LEF-1 protein and destabilize the LEF-1/beta-catenin complex. N-acetyl-L-cysteine (NAC), which can react with the alpha, beta-unsaturated ketone in EA, but not other anti-oxidants, prevented the drug's inhibition of Wnt/beta-catenin activation and its ability to induce apoptosis in CLL cells. CONCLUSIONS/SIGNIFICANCE: Our studies indicate that EA selectively suppresses CLL survival due to inhibition of Wnt/beta-catenin signaling. Antagonizing Wnt signaling in CLL with EA or related drugs may represent an effective treatment of this disease. PMID- 20011539 TI - Erk1 and Erk2 regulate endothelial cell proliferation and migration during mouse embryonic angiogenesis. AB - Angiogenesis is a complex process orchestrated by both growth factors and cell adhesion and is initiated by focal degradation of the vascular basement membrane with subsequent migration and proliferation of endothelial cells. The Ras/Raf/MEK/ERK pathway is required for EC function during angiogenesis. Although in vitro studies implicate ERK1 and ERK2 in endothelial cell survival, their precise role in angiogenesis in vivo remains poorly defined. Cre/loxP technology was used to inactivate Erk1 and Erk2 in endothelial cells during murine development, resulting in embryonic lethality due to severely reduced angiogenesis. Deletion of Erk1 and Erk2 in primary endothelial cells resulted in decreased cell proliferation and migration, but not in increased apoptosis. Expression of key cell cycle regulators was diminished in the double knockout cells, and decreased DNA synthesis could be observed in endothelial cells during embryogenesis. Interestingly, both Paxillin and Focal Adhesion Kinase were expressed at lower levels in endothelial cells lacking Erk1 and Erk2 both in vivo and in vitro, leading to defects in the organization of the cytoskeleton and in cell motility. The regulation of Paxillin and Focal Adhesion Kinase expression occurred post-transcriptionally. These results demonstrate that ERK1 and ERK2 coordinate endothelial cell proliferation and migration during angiogenesis. PMID- 20011540 TI - The extraction of depth structure from shading and texture in the macaque brain. AB - We used contrast-agent enhanced functional magnetic resonance imaging (fMRI) in the alert monkey to map the cortical regions involved in the extraction of 3D shape from the monocular static cues, texture and shading. As in the parallel human imaging study, we contrasted the 3D condition to several 2D control conditions. The extraction of 3D shape from texture (3D SfT) involves both ventral and parietal regions, in addition to early visual areas. Strongest activation was observed in CIP, with decreasing strength towards the anterior part of the intraparietal sulcus (IPS). In the ventral stream 3D SfT sensitivity was observed in a ventral portion of TEO. The extraction of 3D shape from shading (3D SfS) involved predominantly ventral regions, such as V4 and a dorsal potion of TEO. These results are similar to those obtained earlier in human subjects and indicate that the extraction of 3D shape from texture is performed in both ventral and dorsal regions for both species, as are the motion and disparity cues, whereas shading is mainly processed in the ventral stream. PMID- 20011541 TI - FMRI effective connectivity and TMS chronometry: complementary accounts of causality in the visuospatial judgment network. AB - BACKGROUND: While traditionally quite distinct, functional neuroimaging (e.g. functional magnetic resonance imaging: fMRI) and functional interference techniques (e.g. transcranial magnetic stimulation: TMS) increasingly address similar questions of functional brain organization, including connectivity, interactions, and causality in the brain. Time-resolved TMS over multiple brain network nodes can elucidate the relative timings of functional relevance for behavior ("TMS chronometry"), while fMRI functional or effective connectivity (fMRI EC) can map task-specific interactions between brain regions based on the interrelation of measured signals. The current study empirically assessed the relation between these different methods. METHODOLOGY/PRINCIPAL FINDINGS: One group of 15 participants took part in two experiments: one fMRI EC study, and one TMS chronometry study, both of which used an established cognitive paradigm involving one visuospatial judgment task and one color judgment control task. Granger causality mapping (GCM), a data-driven variant of fMRI EC analysis, revealed a frontal-to-parietal flow of information, from inferior/middle frontal gyrus (MFG) to posterior parietal cortex (PPC). FMRI EC-guided Neuronavigated TMS had behavioral effects when applied to both PPC and to MFG, but the temporal pattern of these effects was similar for both stimulation sites. At first glance, this would seem in contradiction to the fMRI EC results. However, we discuss how TMS chronometry and fMRI EC are conceptually different and show how they can be complementary and mutually constraining, rather than contradictory, on the basis of our data. CONCLUSIONS/SIGNIFICANCE: The findings that fMRI EC could successfully localize functionally relevant TMS target regions on the single subject level, and conversely, that TMS confirmed an fMRI EC identified functional network to be behaviorally relevant, have important methodological and theoretical implications. Our results, in combination with data from earlier studies by our group (Sack et al., 2007, Cerebral Cortex), lead to informed speculations on complex brain mechanisms, and TMS disruption thereof, underlying visuospatial judgment. This first in-depth empirical and conceptual comparison of fMRI EC and TMS chronometry thereby shows the complementary insights offered by the two methods. PMID- 20011542 TI - Activin signaling in microsatellite stable colon cancers is disrupted by a combination of genetic and epigenetic mechanisms. AB - BACKGROUND: Activin receptor 2 (ACVR2) is commonly mutated in microsatellite unstable (MSI) colon cancers, leading to protein loss, signaling disruption, and larger tumors. Here, we examined activin signaling disruption in microsatellite stable (MSS) colon cancers. METHODS: Fifty-one population-based MSS colon cancers were assessed for ACVR1, ACVR2 and pSMAD2 protein. Consensus mutation-prone portions of ACVR2 were sequenced in primary cancers and all exons in colon cancer cell lines. Loss of heterozygosity (LOH) was evaluated for ACVR2 and ACVR1, and ACVR2 promoter methylation by methylation-specific PCR and bisulfite sequencing and chromosomal instability (CIN) phenotype via fluorescent LOH analysis of 3 duplicate markers. ACVR2 promoter methylation and ACVR2 expression were assessed in colon cancer cell lines via qPCR and IP-Western blots. Re-expression of ACVR2 after demethylation with 5-aza-2'-deoxycytidine (5-Aza) was determined. An additional 26 MSS colon cancers were assessed for ACVR2 loss and its mechanism, and ACVR2 loss in all tested cancers correlated with clinicopathological criteria. RESULTS: Of 51 MSS colon tumors, 7 (14%) lost ACVR2, 2 (4%) ACVR1, and 5 (10%) pSMAD2 expression. No somatic ACVR2 mutations were detected. Loss of ACVR2 expression was associated with LOH at ACVR2 (p<0.001) and ACVR2 promoter hypermethylation (p<0.05). ACVR2 LOH, but not promoter hypermethylation, correlated with CIN status. In colon cancer cell lines with fully methylated ACVR2 promoter, loss of ACVR2 mRNA and protein expression was restored with 5-Aza treatment. Loss of ACVR2 was associated with an increase in primary colon cancer volume (p<0.05). CONCLUSIONS: Only a small percentage of MSS colon cancers lose expression of activin signaling members. ACVR2 loss occurs through LOH and ACVR2 promoter hypermethylation, revealing distinct mechanisms for ACVR2 inactivation in both MSI and MSS subtypes of colon cancer. PMID- 20011543 TI - Cellular electron microscopy imaging reveals the localization of the Hfq protein close to the bacterial membrane. AB - BACKGROUND: Hfq is a bacterial protein involved in several aspects of nucleic acid transactions, but one of its best-characterized functions is to affect the post-transcriptional regulation of mRNA by virtue of its interactions with stress related small regulatory (sRNA). METHODOLOGY AND PRINCIPAL FINDING: By using cellular imaging based on the metallothionein clonable tag for electron microscopy, we demonstrate here that in addition to its localization in the cytoplasm and in the nucleoid, a significant amount of Hfq protein is located at the cell periphery. Simultaneous immunogold detection of specific markers strongly suggests that peripheral Hfq is close to the bacterial membrane. Because sRNAs regulate the synthesis of several membrane proteins, our result implies that the sRNA- and Hfq-dependent translational regulation of these proteins takes place in the cytoplasmic region underlying the membrane. CONCLUSIONS: This finding supports the proposal that RNA processing and translational machineries dedicated to membrane protein translation may often be located in close proximity to the membrane of the bacterial cell. PMID- 20011544 TI - Impact of previous virological treatment failures and adherence on the outcome of antiretroviral therapy in 2007. AB - BACKGROUND: Combination antiretroviral treatment (cART) has been very successful, especially among selected patients in clinical trials. The aim of this study was to describe outcomes of cART on the population level in a large national cohort. METHODS: Characteristics of participants of the Swiss HIV Cohort Study on stable cART at two semiannual visits in 2007 were analyzed with respect to era of treatment initiation, number of previous virologically failed regimens and self reported adherence. Starting ART in the mono/dual era before HIV-1 RNA assays became available was counted as one failed regimen. Logistic regression was used to identify risk factors for virological failure between the two consecutive visits. RESULTS: Of 4541 patients 31.2% and 68.8% had initiated therapy in the mono/dual and cART era, respectively, and been on treatment for a median of 11.7 vs. 5.7 years. At visit 1 in 2007, the mean number of previous failed regimens was 3.2 vs. 0.5 and the viral load was undetectable (<50 copies/ml) in 84.6% vs. 89.1% of the participants, respectively. Adjusted odds ratios of a detectable viral load at visit 2 for participants from the mono/dual era with a history of 2 and 3, 4, >4 previous failures compared to 1 were 0.9 (95% CI 0.4-1.7), 0.8 (0.4 1.6), 1.6 (0.8-3.2), 3.3 (1.7-6.6) respectively, and 2.3 (1.1-4.8) for >2 missed cART doses during the last month, compared to perfect adherence. From the cART era, odds ratios with a history of 1, 2 and >2 previous failures compared to none were 1.8 (95% CI 1.3-2.5), 2.8 (1.7-4.5) and 7.8 (4.5-13.5), respectively, and 2.8 (1.6-4.8) for >2 missed cART doses during the last month, compared to perfect adherence. CONCLUSIONS: A higher number of previous virologically failed regimens, and imperfect adherence to therapy were independent predictors of imminent virological failure. PMID- 20011545 TI - A comparative analysis of gene expression patterns and cell phenotypes between cervical and peripheral blood mononuclear cells. AB - Studies of the immunological environment in the female genital tract (FGT) are critical for the development of vaccines or microbicides to halt the spread of sexually transmitted infections. Challenges arise due to the difficulties of sampling from this site, and the majority of studies have been conducted utilising peripheral blood mononuclear cells. Identifying functional differences between immune cells of the FGT and peripheral blood would aid in our understanding of mucosal immunology. We compared the gene expression profile of mononuclear cells at these two sites. Messenger RNA expression analysis was performed using gene expression arrays on matched cervical mononuclear cells and peripheral blood mononuclear cells. Further cellular phenotyping was done by 10 colour flow cytometry. Of the 22,185 genes expressed by these samples, 5345 genes were significantly differentially expressed between the cell populations. Most differences can be explained by significantly lower levels of T and B cells and higher levels of macrophages and dendritic cells in the FGT compared with peripheral blood. Several immunologically relevant pathways such as apoptosis and innate immune signalling, and a variety of cytokines and cytokine receptors were differentially expressed. This study highlights the importance of the unique immunological environment of the FGT and identifies important differences between systemic and mucosal immune compartments. PMID- 20011546 TI - Recruitment of Rad51 and Rad52 to short telomeres triggers a Mec1-mediated hypersensitivity to double-stranded DNA breaks in senescent budding yeast. AB - Telomere maintenance is required for chromosome stability, and telomeres are typically replicated by the action of telomerase. In both mammalian tumor and yeast cells that lack telomerase, telomeres are maintained by an alternative recombination mechanism. Here we demonstrated that the budding yeast Saccharomyces cerevisiae type I survivors derived from telomerase-deficient cells were hypersensitive to DNA damaging agents. Assays to track telomere lengths and drug sensitivity of telomerase-deficient cells from spore colonies to survivors suggested a correlation between telomere shortening and bleomycin sensitivity. Our genetic studies demonstrated that this sensitivity depends on Mec1, which signals checkpoint activation, leading to prolonged cell-cycle arrest in senescent budding yeasts. Moreover, we also observed that when cells equipped with short telomeres, recruitments of homologous recombination proteins, Rad51 and Rad52, were reduced at an HO-endonuclease-catalyzed double-strand break (DSB), while their associations were increased at chromosome ends. These results suggested that the sensitive phenotype may be attributed to the sequestration of repair proteins to compromised telomeres, thus limiting the repair capacity at bona fide DSB sites. PMID- 20011547 TI - Nucleotide, cytogenetic and expression impact of the human chromosome 8p23.1 inversion polymorphism. AB - BACKGROUND: The human chromosome 8p23.1 region contains a 3.8-4.5 Mb segment which can be found in different orientations (defined as genomic inversion) among individuals. The identification of single nucleotide polymorphisms (SNPs) tightly linked to the genomic orientation of a given region should be useful to indirectly evaluate the genotypes of large genomic orientations in the individuals. RESULTS: We have identified 16 SNPs, which are in linkage disequilibrium (LD) with the 8p23.1 inversion as detected by fluorescent in situ hybridization (FISH). The variability of the 8p23.1 orientation in 150 HapMap samples was predicted using this set of SNPs and was verified by FISH in a subset of samples. Four genes (NEIL2, MSRA, CTSB and BLK) were found differentially expressed (p<0.0005) according to the orientation of the 8p23.1 region. Finally, we have found variable levels of mosaicism for the orientation of the 8p23.1 as determined by FISH. CONCLUSION: By means of dense SNP genotyping of the region, haplotype-based computational analyses and FISH experiments we could infer and verify the orientation status of alleles in the 8p23.1 region by detecting two short haplotype stretches at both ends of the inverted region, which are likely the relic of the chromosome in which the original inversion occurred. Moreover, an impact of 8p23.1 inversion on gene expression levels cannot be ruled out, since four genes from this region have statistically significant different expression levels depending on the inversion status. FISH results in lymphoblastoid cell lines suggest the presence of mosaicism regarding the 8p23.1 inversion. PMID- 20011548 TI - Access to vaccines: A call to action. PMID- 20011549 TI - Vaccine schedules. PMID- 20011550 TI - The child with developmental delay: An approach to etiology. AB - OBJECTIVE: To describe an approach to history, physical examination and investigation for the developmentally delayed child. METHODS: A review of electronic databases from 1997 to 2001 was done searching for articles relating to the approach to or investigations of children with developmental delay. Five studies, including a review of a consensus conference on evaluation of mental retardation, were chosen because of their general approaches to developmental delay and/or mental retardation, or specific evaluations of a particular laboratory investigation. CONCLUSIONS: A diagnosis or cause of mental retardation can be identified in 20% to 60% of cases. Evaluation of the developmentally delayed child should include a detailed history and physical examination, taking special care to record a three-generation pedigree, as well as to look for dysmorphic features. If no other cause is apparent, routine investigations should include a chromosome study and fragile X studies. Further investigations are warranted depending on the clinical features. PMID- 20011551 TI - Practical aspects of genetic testing in ambulatory paediatrics and child health care. AB - BACKGROUND: An overview of practical aspects of genetic testing in ambulatory practice in Canada. OBJECTIVE: To provide an overview and practical references for child health specialists and paediatricians on genetic services that can be provided to their patients. METHODS/RESULTS: A review and summary of areas of genetic testing options available across Canada. CONCLUSIONS: Modern Canadian paediatric practice requires office-based practitioners to fully understand and utilize genetics testing in their offices. PMID- 20011552 TI - Prenatal diagnosis for paediatricians. AB - In Ontario, approximately 140,000 women deliver newborn infants each year. Of these women, 60,000 to 70,000 have multiple marker screening, 10,000 undergo amniocentesis or chorion villus sampling and virtually all have at least one prenatal ultrasound. Multiple marker screening is not used in every province and territory; however, amniocentesis and prenatal ultrasound are used throughout Canada. Most paediatric patients will have been exposed to some form of prenatal diagnosis. If an abnormality is found prenatally, parents may have concerns to discuss with the paediatrician after the child is born. Likewise, if a child with a problem is born following a normal pregnancy, the parents will want to know why the problem was missed prenatally. Paediatricians should be aware of prenatal tests that have been performed to understand better their patients and their families. PMID- 20011553 TI - The family history: An integral component of paediatric health assessment. AB - Conducting the paediatric health history is a vital skill for the paediatrician. However, published guidelines on this topic often fail to emphasize the value that the family history adds to paediatric health assessment. For the healthy child, the family history may reveal risk factors for juvenile and/or adult-onset conditions, the adverse effects of which may be ameliorated with medical intervention or surveillance. For the child with a health problem, a detailed family history may help to guide the clinician through the diagnostic and treatment processes. PMID- 20011554 TI - Genetic issues in the care of the adolescent patient. AB - Genetic issues are important in the primary care of adolescents. A genetic diagnosis may not be made until adolescence, when the teenager presents with the first signs or symptoms of the condition. The physician's knowledge of the natural history of a genetic disease will aid in the anticipatory guidance for teens and their parents. The physician may be called upon to advise the patient regarding hormone therapy or contraception. The paediatrician may initiate topics such as sexuality and sex education for the developmentally delayed patient. The paediatrician is also the advocate for the teenager, who must gain independence from the family in medical as well as other aspects of life. This article examines some of these issues, using cases to illustrate the genetic problems and approaches in the care of the teenaged patient. PMID- 20011556 TI - Frostbite. PMID- 20011555 TI - Guidelines for genetic testing of healthy children. PMID- 20011557 TI - Surveillance of rare genetic disorders: No longer orphan diseases. PMID- 20011560 TI - Accomplishments in 2008 in the adjuvant treatment of colon cancer. AB - Overview of the Disease IncidencePrognosisCurrent General Therapy Standards StagingLymph Node ExcisionStage III DiseaseStage II DiseaseMolecular MarkersAccomplishments During the Year Therapy Cytotoxic ChemotherapyChemotherapy Plus Targeted TherapySpecial PopulationsBasic Science-BiomarkersMethodologyWhat Needs to Be Done Application of the AccomplishmentsControversies and DisagreementsFuture Directions Comments on ResearchObstacles to Progress. PMID- 20011559 TI - Accomplishments in 2008 in the management of curable metastatic colorectal cancer. AB - Overview of the Disease IncidencePrognosisCurrent Therapy Standards Colorectal Liver Metastases (CRLM) Resectable TumorsStrategies to Convert Nonresectable Liver Metastases to Resectable StatusSynchronous Colorectal Liver MetastasesPredictors of Survival After Resection of CRLMPeritoneal Carcinomatosis (PC) From Colorectal CancerColorectal Pulmonary Metastases (CRPM)Colorectal Liver Metastases With Extrahepatic DiseaseAccomplishments (or Lack of Accomplishments) During the Year Therapy New Staging SystemSystemic Chemotherapy in Resectable Liver MetastasesSystemic Chemotherapy in Nonresectable Liver MetastasesSelective Internal Radiation Therapy (SIRT)Selection of Patients for Liver ResectionRadiofrequency AblationBiomarkersWhat Needs To Be Done? Optimizing Patient CareFuture Directions Comments on ResearchObstacles to Overcome. PMID- 20011561 TI - Accomplishments in 2008 in the treatment of advanced metastatic colorectal cancer. AB - Overview of the Disease IncidencePredictive BiomarkersCurrent General Therapy StandardsAccomplishments During the Year Combined Modality Management of Liver MetastasesTreatment of Advanced Disease Equivalence of FOLFOX and CAPOXAddition of Bevacizumab to Standard ChemotherapyDual EGFR and VEGF InhibitionBevacizumab Upon Progression While on First-Line TherapyBenefit in the Elderly PopulationWhat Needs to be Done Comments on ResearchObstacles to Progress. PMID- 20011562 TI - Accomplishments in 2008 in the management of hepatobiliary cancers. AB - HEPATOCELLULAR CARCINOMA: EpidemiologyManagement of HCC-Standard Approaches and Reports During the Past Year SurgeryLocoregional Treatment Local AblationChemoembolizationOther Local Treatment ModalitiesSystemic TreatmentClinical Trial DesignBasic Science and Biomarkers Basic and Translational SciencePrognostic and Predictive Markers Prognostic MarkersPredictive MarkersAccomplishments and Future DirectionsApplication of the AccomplishmentsFuture Directions BILIARY TRACT CANCERS: Overview of the DiseaseManagement of Biliary Tract Cancers-Current Approaches and Reports During the Past Year Surgical Resection and Adjuvant TherapySystemic Therapy for Unresectable or Metastatic Biliary Tract Cancer Cytotoxic ChemotherapyCytotoxic Chemotherapy in Combination With Targeted AgentSingle-Agent Targeted TherapyCombination of Targeted AgentsBiologyFuture Directions. PMID- 20011563 TI - Accomplishments in 2008 in the management of localized pancreatic cancer. AB - Overview of the DiseaseIncidenceRisk FactorsPrognostic or Predictive FactorsCurrent General Therapy StandardsAccomplishments During the YearTherapyBiomarkersBasic ScienceWhat Needs to Be Done?Applications of the AccomplishmentsControversies and DisagreementsFuture DirectionsComments on ResearchObstacles to Progress. PMID- 20011564 TI - Accomplishments in 2008 in the treatment of metastatic pancreatic cancer. AB - Overview of the DiseaseIncidencePrognosisCurrent Treatment StandardsCytotoxic TherapyTargeted TherapySalvage TherapyAccomplishments or Lack of AccomplishmentsTherapySalvage TherapyBiomarkersApplication of the AccomplishmentsFuture DirectionsCurrent Basic ResearchOngoing Clinical StudiesMajor Obstacles to Progress in Drug DevelopmentGemcitabine PlatformChoice of Molecular TargetsPatient Selection. PMID- 20011565 TI - Accomplishments in 2008 in the management of localized gastric cancer. AB - Overview of the Disease ProcessIncidencePrognostic or Predictive FactorsCurrent General Therapy StandardsSurgeryAdjuvant ChemoradiationPerioperative ChemotherapyAdjuvant Oral Fluoropyrimidine MonotherapyAccomplishments in 2008TherapyBiomarkers and Basic ScienceWhat Needs to Be DoneApplications of the AccomplishmentsControversies and DisagreementsSpecial PopulationsCurrent and Future DirectionsComments on ResearchObstacles to Progress. PMID- 20011566 TI - Accomplishments in 2008 in the management of esophageal cancer. AB - Overview of the Disease ProcessIncidencePrognostic or Predictive FactorsCurrent General Therapy StandardsSurgeryCombined Treatment ModalitiesAccomplishments in 2008TherapyBiomarkers & Basic ScienceWhat Needs to be DoneApplications of the AccomplishmentsControversies and DisagreementsSpecial PopulationsFuture DirectionsComments on ResearchObstacles to Progress. PMID- 20011567 TI - Accomplishments in 2008 in the management of advanced gastroesophageal cancer. AB - Overview of the Disease Process IncidencePrognosisPrognostic or Predictive FactorsCurrent General Therapy Standards Regional VariationsEvidence for Individual AgentsAccomplishments in 2008 TherapyBiomarkersBasic ScienceWhat Needs to be Done Applications of the AccomplishmentsControversies and DisagreementsSpecial PopulationsFuture Directions Comments on ResearchObstacles to Progress. PMID- 20011568 TI - Accomplishments in 2008 in the management of gastrointestinal neuroendocrine tumors. AB - Overview of the Disease ProcessIncidencePrognostic FactorsGeneral Therapy StandardsAccomplishments and Lack of Accomplishments During the YearSomatostatin AnalogsPeptide Receptor-Targeted TherapySelective Internal Radiotherapy (SIRT)CYTOTOXIC THERAPIES: TemozolomideVEGF Pathway InhibitorsmTOR InhibitorsDevelopment of BiomarkersWhat Needs to be Done (Application of the Accomplishments)Controversies and DisagreementsHistologic Classification and Staging of Neuroendocrine TumorsClinical Trial Design and End PointsFuture Directions. PMID- 20011569 TI - Accomplishments in 2008 in the management of gastrointestinal stromal tumors. AB - Overview of the Disease ProcessIncidencePrognosisPredictive MarkersCurrent General Therapy Standards in North America and EuropeLocalized or Potentially Resectable DiseaseUnresectable or Metastatic DiseaseAccomplishments During the YearTherapySurgical Issues and Perioperative TherapyImatinibSunitinibNew DrugsBiomarkersBasic and Other Translational ScienceWhat Needs to Be DoneFuture DirectionsComments on ResearchObstacles to Progress. PMID- 20011570 TI - Accomplishments in 2008 in biologic markers for gastrointestinal cancers-focus on colorectal cancer. AB - OverviewPredictive/Prognostic FactorsKRAS StatusLigands of EGFRPhosphoinositide 3 kinases (PI3Ks)Phosphatase and Tensin Homolog (PTEN)Microsatellite Instability (MSI)Inflammatory Bowel Disease (IBD) and Colon CancerFlat and Polypoid DysplasiaOther Non-Morphology-Based Predictive Markers of Cancer in IBDPotential Applications in Clinical PracticeTargeting TherapyFuture Directions. PMID- 20011571 TI - Accomplishments in gastrointestinal oncology in 2008: conclusion. PMID- 20011573 TI - For each question, choose the correct answer from the multiple-choice list. PMID- 20011572 TI - Accomplishments in 2008 in the adjuvant treatment of rectal cancer. AB - Overview of the Disease IncidencePrognostic FactorsCurrent General Therapy Standards Surgery(Neo) Adjuvant Radiotherapy and ChemotherapyAccomplishments During the Year Time Duration Between Radiotherapy and SurgeryHistopathology Post Neoadjuvant TherapyMetabolic Imaging During Neoadjuvant TherapyNew Drugs in Combination with RadiotherapyNew StrategiesFuture Strategies ControversiesComments on ResearchObstacles to Progress. PMID- 20011574 TI - Treatment of bipolar disorder. AB - In this article, we investigate the range of treatments prescribed for bipolar disorder. Our analysis shows that, while a large portion of patients is treated by a single mechanism of action (44%), an equally sizable group of patients receives two or more drug classes (56%) to treat the disorder. From a therapeutic class perspective, 71 percent of patients with bipolar disorder receive an atypical antipsychotic, 53 percent receive a mood stabilizer, and 30 percent receive an antidepressant. While antipsychotics and mood stabilizers represent the vast majority of bipolar disorder monotherapy (90%), antidepressants are more commonly seen as part of a combination treatment. PMID- 20011575 TI - Borderline personality and criminality. AB - Borderline personality disorder is characteristically associated with a broad variety of psychiatric symptoms and aberrant behaviors. In this edition of The Interface, we discuss the infrequently examined association between borderline personality disorder and criminality. According to our review of the literature, in comparison with the rates of borderline personality disorder encountered in the general population, borderline personality disorder is over-represented in most studies of inmates. At the same time, there is considerable variation in the reported rates of this Axis II disorder in prison populations, which may be attributed to the methodologies of and populations in the various studies. Overall, female criminals appear to exhibit higher rates of borderline personality disorder, and it is oftentimes associated with a history of childhood sexual abuse, perpetration of impulsive and violent crimes, comorbid antisocial traits, and incarceration for domestic violence. PMID- 20011576 TI - Decreased Use of Antidepressants in Youth After US Food and Drug Administration Black Box Warning. AB - Objective. This study evaluates changes in use of antidepressants in children and adolescents after the US Food and Drug Administration black box warning for increased risk of suicide.Method. A retrospective chart review was completed for children and adolescents (ages 4-17) who were diagnosed with depressive or anxiety disorders in an outpatient clinic and offered a trial of antidepressants between September 2003 and February 2004 (before the black box warning) and between January 2005 and June 2005 (after the black box warning). Statistical analyses were performed with the SPSS version 17 and R package version 2.9.1. Univariate analysis was conducted using the Fisher's Exact test.Results. The odds ratio calculated for the different groups suggests that in all the groups, the proportion of acceptance of antidepressant use was greater before the black box warning as compared to after the black box warning (odds ratio>1). It was also found that upon combining the age groups after the warning and comparing them, based on the diagnoses, there was a greater degree of refusal of antidepressant therapy when a diagnosis of anxiety disorder was made as compared to a diagnosis of depressive disorder (p=0.017).Conclusion. There has been a decrease in the use of antidepressant therapy in children and adolescents following the US Food and Drug Administration black box warning for risk of suicide. A limitation of this study is that reasons for refusal of antidepressent therapy by parents or guardians of children and adolescents were not collected; therefore, there is no certainty that the black box warning was the primary reason for refusal. PMID- 20011577 TI - Near-death experiences and psychotherapy. AB - Psychiatrists are likely to come into contact with patients who have had near death experiences, who may have a variety of reactions to the experience, and who may benefit from psychotherapy. We may also have opportunities to work with individuals who are reacting to others who have had such experiences. There is much a psychiatrist can offer to these people, including listening respectfully, being nonjudgemental, normalizing the experience, providing education, and assisting with integrating the experience into their lives to develop or maintain the best possible functioning. PMID- 20011579 TI - Wake up america!: and keep this country beautiful. PMID- 20011578 TI - Direct measurement of disability. AB - Schizophrenia is associated with substantial disability in everyday functioning. Environmental factors, such as disability compensation and opportunities, impact on the ability of people with the illness to perform real-world activities, particularly those that are vocational in nature. Measures of abilities to perform these functions have been developed that measure competence in social, everyday living, and vocational domains. These indices have measurement characteristics consistent with those of neuropsychological tests, indicating that they have the potential to be valid measures of the ability to perform in real-world functional situations. It is our belief that these measures will be widely employed in later research on functioning in schizophrenia, and many of these tests have the potential to be used in clinical settings. PMID- 20011580 TI - A Simple and Practical Spreadsheet-Based Method to Extract Single-Molecule Dissociation Kinetics from Variable Loading-Rate Force Spectroscopy Data. PMID- 20011581 TI - Towards the stereoselective synthesis of alpha-methylated (2S,3aS,7aS) octahydroindole-2-carboxylic acid. AB - A high yielding and remarkably stereoselective alpha-methylation reaction of the (2S,3aS,7aS) stereoisomer of octahydroindole-2-carboxylic acid, (S,S,S)-Oic, suitably protected is described. The severe steric hindrance imposed by the fused cyclohexane ring, which prevents the application of Seebach's self-reproduction of chirality methodology, accounts for the formation of (S,S,S)-(alphaMe)Oic with high selectivity and retention of configuration. PMID- 20011582 TI - AlloDerm Sling for Correction of Synmastia After Immediate, Tissue Expander, Breast Reconstruction in Thin Women. AB - INTRODUCTION: Synmastia is a condition in which the breasts are conjoint and the natural intermammary sulcus is obliterated. It is the rarest type of breast implant malpositioning during breast augmentation; however, it is the most difficult one to correct. AlloDerm is an acellular dermal matrix that is assuming a major role in immediate breast reconstruction in recent years. METHODS: In the past 2 years, we have treated 3 thin women, a total of 6 breasts, for correction of synmastia after bilateral immediate breast reconstruction, using tissue expanders and skin sparing mastectomy. All of them suffered from synmastia, which manifested immediately after the mastectomy and accelerated during tissue expander inflation. We exchanged the expander into silicone implants, and during the same procedure we corrected the synmastia, using an AlloDerm sling. A thick sheet of AlloDerm (Life-Cell Corp, Branchbung, NJ) is used and the AlloDerm sheet is designed into a long narrow sling. Then, the sling is sutured into place. RESULTS: This technique successfully resolved the synmastia. CONCLUSION: The use of an AlloDerm sling to reinforce the capsule and the AlloDerm incorporation into it ensures a sound solution with a low recurrence rate. PMID- 20011583 TI - Vacuum-assisted closure of perineal war wound related to rectum. AB - INTRODUCTION: Vacuum-assisted wound closure therapy has widely been used in various clinical applications with successful results and has considerably increased in popularity over the past decade. The patient who sustained a complex war wound to his perineum has been presented. METHODS: After the initial treatment he was discharged from the hospital in which he had been treated for 4 days in Iraq. On the examination, all wounds were deeply contaminated with foreign bodies and also involved significant volume of devitalized tissue. Perineal injury had not only caused a large skin defect but also left a deep wound leading to rectal perforation, so the wound and its borders were quite contaminated and infected by rectal contents. After improving his general condition with medical treatment, he underwent an immediate operation in which first a colostomy was performed and then the wound tract placed between perineum and rectum was sharply and extensively debrided to viable-appearing bleeding tissue to remove the whole necrotic tissues, foreign bodies, cloths, and debris. At the end of the intervention, a negative pressure dressing was applied and used during 12 days and then completed. RESULTS: The wound tract obliterated entirely without permitting any leakage of rectal contents, and wound bed appeared clean, granulated, contracted, and viable enough for definitive closure with flap mobilization. CONCLUSIONS: When dealing with this experience presenting an unusual wound that was in a very difficult area of the body for the treatment, perineum, caused from a challenging reason, war injury, and also was complicated with rectal injury, the technique seems to have a significant beneficial effect on the healing of complicated wounds such as in perineal wound and war wound, even if these are at risk of severe infection and progressive tissue necrosis. PMID- 20011584 TI - False positive rate of rapid oral fluid HIV tests increases as kits near expiration date. AB - BACKGROUND: Because a recent cluster of false positive results on the OraQuick ADVANCE Rapid HIV-1/2 Antibody Test occurred in San Francisco on test kits close to their expiration date, we decided to assess the relationship between time to expiration and rate of false positive results from tests used with oral fluid. METHODOLOGY/PRINCIPAL FINDINGS: We analyzed results of 20,904 tests with either an initial HIV-negative result (n = 20,828) or a preliminary positive result that was then negative on confirmatory tests (n = 76). We computed specificity for kits with time to expiration from < or = 1 to > or = 6 months, with exact binomial confidence intervals, then used logistic regression to estimate the independent association of time to expiration with false positive results, adjusting for site and technician effects. For 1,108 kits used in the last month before expiration, specificity was 98.83% (95% exact binomial confidence interval (CI) 98.00%-99.37%); the upper bound is below the claimed specificity of 99.60%. After adjustment using regression standardization for the effects of site, test lot, and technician factors, adjusted specificity in the last month before expiration was 99.18% (95% bootstrap confidence interval 98.60-99.57%). CONCLUSIONS/SIGNIFICANCE: We found that specificity of the OraQuick ADVANCE with oral fluid declined significantly with < or = 1 month remaining to expiration, leaving little margin for error from other sources. PMID- 20011585 TI - Estradiol stimulates vasodilatory and metabolic pathways in cultured human endothelial cells. AB - Vascular effects of estradiol are being investigated because there are controversies among clinical and experimental studies. DNA microarrays were used to investigate global gene expression patterns in cultured human umbilical vein endothelial cells (HUVEC) exposed to 1 nmol/L estradiol for 24 hours. When compared to control, 187 genes were identified as differentially expressed with 1.9-fold change threshold. Supervised principal component analysis and hierarchical cluster analysis revealed the differences between control and estradiol-treated samples. Physiological concentrations of estradiol are sufficient to elicit significant changes in HUVEC gene expression. Notch signaling, actin cytoskeleton signaling, pentose phosphate pathway, axonal guidance signaling and integrin signaling were the top-five canonical pathways significantly regulated by estrogen. A total of 26 regulatory networks were identified as estrogen responsive. Microarray data were confirmed by quantitative RT-PCR in cardiovascular meaning genes; cyclooxygenase (COX)1, dimethylarginine dimethylaminohydrolase (DDAH)2, phospholipase A2 group IV (PLA2G4) B, and 7 dehydrocholesterol reductase were up-regulated by estradiol in a dose-dependent and estrogen receptor-dependent way, whereas COX2, DDAH1 and PLA2G4A remained unaltered. Moreover, estradiol-induced COX1 gene expression resulted in increased COX1 protein content and enhanced prostacyclin production. DDAH2 protein content was also increased, which in turn decreased asymmetric dimethylarginine concentration and increased NO release. All stimulated effects of estradiol on gene and protein expression were estrogen receptor-dependent, since were abolished in the presence of the estrogen receptor antagonist ICI 182780. This study identifies new vascular mechanisms of action by which estradiol may contribute to a wide range of biological processes. PMID- 20011586 TI - A macaque model to study vaginal HSV-2/immunodeficiency virus co-infection and the impact of HSV-2 on microbicide efficacy. AB - BACKGROUND: Herpes simplex virus type-2 (HSV-2) infection enhances the transmission and acquisition of human immunodeficiency virus (HIV). This occurs in symptomatic and asymptomatic stages of HSV-2 infection, suggesting that obvious herpetic lesions are not required to increase HIV spread. An animal model to investigate the underlying causes of the synergistic action of the two viruses and where preventative strategies can be tested under such complex physiological conditions is currently unavailable. METHODOLOGY/PRINCIPAL FINDINGS: We set out to establish a rhesus macaque model in which HSV-2 infection increases the susceptibility to vaginal infection with a model immunodeficiency virus (simian human immunodeficiency virus, SHIV-RT), and to more stringently test promising microbicides. HSV-2 exposure significantly increased the frequency of vaginal SHIV-RT infection (n = 6). Although cervical lesions were detected in only approximately 10% of the animals, long term HSV-2 DNA shedding was detected (in 50% of animals followed for 2 years). Vaginal HSV-2 exposure elicited local cytokine/chemokine (n = 12) and systemic low-level HSV-2-specific adaptive responses in all animals (n = 8), involving CD4(+) and CD8(+) HSV-specific T cells (n = 5). Local cytokine/chemokine responses were lower in co-infected animals, while simian immunodeficiency virus (SIV)-specific adaptive responses were comparable in naive and HSV-2-infected animals (n = 6). Despite the increased frequency of SHIV-RT infection, a new generation microbicide gel, comprised of Carraguard(R) and a non-nucleoside reverse transcriptase inhibitor MIV-150 (PC-817), blocked vaginal SHIV-RT infection in HSV-2-exposed animals (n = 8), just as in naive animals. CONCLUSIONS/SIGNIFICANCE: We established a unique HSV-2 macaque model that will likely facilitate research to define how HSV-2 increases HIV transmission, and enable more rigorous evaluation of candidate anti viral approaches in vivo. PMID- 20011587 TI - RNA editing genes associated with extreme old age in humans and with lifespan in C. elegans. AB - BACKGROUND: The strong familiality of living to extreme ages suggests that human longevity is genetically regulated. The majority of genes found thus far to be associated with longevity primarily function in lipoprotein metabolism and insulin/IGF-1 signaling. There are likely many more genetic modifiers of human longevity that remain to be discovered. METHODOLOGY/PRINCIPAL FINDINGS: Here, we first show that 18 single nucleotide polymorphisms (SNPs) in the RNA editing genes ADARB1 and ADARB2 are associated with extreme old age in a U.S. based study of centenarians, the New England Centenarian Study. We describe replications of these findings in three independently conducted centenarian studies with different genetic backgrounds (Italian, Ashkenazi Jewish and Japanese) that collectively support an association of ADARB1 and ADARB2 with longevity. Some SNPs in ADARB2 replicate consistently in the four populations and suggest a strong effect that is independent of the different genetic backgrounds and environments. To evaluate the functional association of these genes with lifespan, we demonstrate that inactivation of their orthologues adr-1 and adr-2 in C. elegans reduces median survival by 50%. We further demonstrate that inactivation of the argonaute gene, rde-1, a critical regulator of RNA interference, completely restores lifespan to normal levels in the context of adr 1 and adr-2 loss of function. CONCLUSIONS/SIGNIFICANCE: Our results suggest that RNA editors may be an important regulator of aging in humans and that, when evaluated in C. elegans, this pathway may interact with the RNA interference machinery to regulate lifespan. PMID- 20011588 TI - Exome sequencing of a multigenerational human pedigree. AB - Over the next few years, the efficient use of next-generation sequencing (NGS) in human genetics research will depend heavily upon the effective mechanisms for the selective enrichment of genomic regions of interest. Recently, comprehensive exome capture arrays have become available for targeting approximately 33 Mb or approximately 180,000 coding exons across the human genome. Selective genomic enrichment of the human exome offers an attractive option for new experimental designs aiming to quickly identify potential disease-associated genetic variants, especially in family-based studies. We have evaluated a 2.1 M feature human exome capture array on eight individuals from a three-generation family pedigree. We were able to cover up to 98% of the targeted bases at a long-read sequence read depth of > or = 3, 86% at a read depth of > or = 10, and over 50% of all targets were covered with > or = 20 reads. We identified up to 14,284 SNPs and small indels per individual exome, with up to 1,679 of these representing putative novel polymorphisms. Applying the conservative genotype calling approach HCDiff, the average rate of detection of a variant allele based on Illumina 1 M BeadChips genotypes was 95.2% at > or = 10x sequence. Further, we propose an advantageous genotype calling strategy for low covered targets that empirically determines cut off thresholds at a given coverage depth based on existing genotype data. Application of this method was able to detect >99% of SNPs covered > or = 8x. Our results offer guidance for "real-world" applications in human genetics and provide further evidence that microarray-based exome capture is an efficient and reliable method to enrich for chromosomal regions of interest in next-generation sequencing experiments. PMID- 20011589 TI - IFNgamma response to Mycobacterium tuberculosis, risk of infection and disease in household contacts of tuberculosis patients in Colombia. AB - OBJECTIVES: Household contacts (HHCs) of pulmonary tuberculosis patients are at high risk of Mycobacterium tuberculosis infection and early disease development. Identification of individuals at risk of tuberculosis disease is a desirable goal for tuberculosis control. Interferon-gamma release assays (IGRAs) using specific M. tuberculosis antigens provide an alternative to tuberculin skin testing (TST) for infection detection. Additionally, the levels of IFNgamma produced in response to these antigens may have prognostic value. We estimated the prevalence of M. tuberculosis infection by IGRA and TST in HHCs and their source population (SP), and assessed whether IFNgamma levels in HHCs correlate with tuberculosis development. METHODS: A cohort of 2060 HHCs was followed for 2-3 years after exposure to a tuberculosis case. Besides TST, IFNgamma responses to mycobacterial antigens: CFP, CFP-10, HspX and Ag85A were assessed in 7-days whole blood cultures and compared to 766 individuals from the SP in Medellin, Colombia. Isoniazid prophylaxis was not offered to child contacts because Colombian tuberculosis regulations consider it only in children under 5 years, TST positive without BCG vaccination. RESULTS: Using TST 65.9% of HHCs and 42.7% subjects from the SP were positive (OR 2.60, p<0.0001). IFNgamma response to CFP-10, a biomarker of M. tuberculosis infection, tested positive in 66.3% HHCs and 24.3% from the SP (OR = 6.07, p<0.0001). Tuberculosis incidence rate was 7.0/1000 person years. Children <5 years accounted for 21.6% of incident cases. No significant difference was found between positive and negative IFNgamma responders to CFP-10 (HR 1.82 95% CI 0.79-4.20 p = 0.16). However, a significant trend for tuberculosis development amongst high HHC IFNgamma producers was observed (trend Log rank p = 0.007). DISCUSSION: CFP-10-induced IFNgamma production is useful to establish tuberculosis infection prevalence amongst HHC and identify those at highest risk of disease. The high tuberculosis incidence amongst children supports administration of chemoprophylaxis to child contacts regardless of BCG vaccination. PMID- 20011590 TI - Systems-level comparison of host-responses elicited by avian H5N1 and seasonal H1N1 influenza viruses in primary human macrophages. AB - Human disease caused by highly pathogenic avian influenza (HPAI) H5N1 can lead to a rapidly progressive viral pneumonia leading to acute respiratory distress syndrome. There is increasing evidence from clinical, animal models and in vitro data, which suggests a role for virus-induced cytokine dysregulation in contributing to the pathogenesis of human H5N1 disease. The key target cells for the virus in the lung are the alveolar epithelium and alveolar macrophages, and we have shown that, compared to seasonal human influenza viruses, equivalent infecting doses of H5N1 viruses markedly up-regulate pro-inflammatory cytokines in both primary cell types in vitro. Whether this H5N1-induced dysregulation of host responses is driven by qualitative (i.e activation of unique host pathways in response to H5N1) or quantitative differences between seasonal influenza viruses is unclear. Here we used microarrays to analyze and compare the gene expression profiles in primary human macrophages at 1, 3, and 6 h after infection with H5N1 virus or low-pathogenic seasonal influenza A (H1N1) virus. We found that host responses to both viruses are qualitatively similar with the activation of nearly identical biological processes and pathways. However, in comparison to seasonal H1N1 virus, H5N1 infection elicits a quantitatively stronger host inflammatory response including type I interferon (IFN) and tumor necrosis factor (TNF)-alpha genes. A network-based analysis suggests that the synergy between IFN beta and TNF-alpha results in an enhanced and sustained IFN and pro-inflammatory cytokine response at the early stage of viral infection that may contribute to the viral pathogenesis and this is of relevance to the design of novel therapeutic strategies for H5N1 induced respiratory disease. PMID- 20011591 TI - Insufficient milk supply and breast cancer risk: a systematic review. AB - BACKGROUND: An association between insufficient milk supply, the inability of a mother's breast milk to provide sufficiently for her infant, and breast cancer has been suggested by observations in animal models. To determine if an association has been reported in epidemiological studies of human breast cancer, a systematic review of the literature has been conducted. We also sought to identify the methodological limitations of existing studies to guide the design of any future prospective studies in this field. METHODOLOGY/PRINCIPAL FINDINGS: PubMed, EMBASE, Web of Science, BIOSIS, and CAB abstracts were searched. We selected any study that (1) assessed breast cancer in association with breastfeeding history and (2) examined the relationship between insufficient milk supply with breast cancer. Seven relevant studies were identified that met both criteria. There was statistically significant heterogeneity among the results which likely reflects clinically significant differences in definitions of insufficient milk supply and reference groups that were used. Among premenopausal women who had experienced insufficient milk supply, odds ratios (ORs) for breast cancer risk ranged from 0.9 to 16.3. Among postmenopausal women, ORs ranged from 0.6 to 6.7. Based on the range of odds ratios obtained in the studies reported in this review, it remains unclear if there is a true association between insufficient milk supply and breast cancer. CONCLUSIONS/SIGNIFICANCE: Although some studies have shown a strong positive association, there is no consistent evidence for an effect of insufficient milk supply on breast cancer risk. Exposure definitions are in need of improvement in order to focus on primary insufficient milk supply. Reference groups consisting of women who have successfully breastfed may also introduce positive bias (inflation of the odds ratio) into study results because of the protective effect of prolonged breastfeeding in the control group. PMID- 20011592 TI - Immunological memory transferred with CD4 T cells specific for tuberculosis antigens Ag85B-TB10.4: persisting antigen enhances protection. AB - BACKGROUND: High levels of death and morbidity worldwide caused by tuberculosis has stimulated efforts to develop a new vaccine to replace BCG. A number of Mycobacterium tuberculosis (Mtb)-specific antigens have been synthesised as recombinant subunit vaccines for clinical evaluation. Recently a fusion protein of TB antigen Ag85B combined with a second immunodominant TB antigen TB10.4 was emulsified with a novel non-phospholipid-based liposomal adjuvant to produce a new subunit vaccine, investigated here. Currently, there is no consensus as to whether or not long-term T cell memory depends on a source of persisting antigen. To explore this and questions regarding lifespan, phenotype and cytokine patterns of CD4 memory T cells, we developed an animal model in which vaccine-induced CD4 memory T cells could transfer immunity to irradiated recipients. METHODOLOGY/PRINCIPAL FINDINGS: The transfer of protective immunity using Ag85B TB10.4-specific, CD45RB(low) CD62L(low) CD4 T cells was assessed in sub-lethally irradiated recipients following challenge with live BCG, used here as a surrogate for virulent Mtb. Donor T cells also carried an allotype marker allowing us to monitor numbers of antigen-specific, cytokine-producing CD4 T cells in recipients. The results showed that both Ag85B-TB10.4 and BCG vaccination induced immunity that could be transferred with a single injection of 3x10(6) CD4 T cells. Ten times fewer numbers of CD4 T cells (0.3x10(6)) from donors immunised with Ag85B-TB10.4 vaccine alone, transferred equivalent protection. CD4 T cells from donors primed by BCG and boosted with the vaccine similarly transferred protective immunity. When BCG challenge was delayed for 1 or 2 months after transfer (a test of memory T cell survival) recipients remained protected. Importantly, recipients that contained persisting antigen, either live BCG or inert vaccine, showed significantly higher levels of protection (p<0.01). Overall the numbers of IFN-gamma-producing CD4 T cells were poorly correlated with levels of protection. CONCLUSIONS/SIGNIFICANCE: The Ag85B-TB10.4 vaccine, with or without BCG-priming, generated TB-specific CD4 T cells that transferred protective immunity in mice challenged with BCG. The level of protection was enhanced in recipients containing a residual source of specific antigen that could be either viable or inert. PMID- 20011593 TI - Primordial germ cell-like cells differentiated in vitro from skin-derived stem cells. AB - BACKGROUND: We have previously demonstrated that stem cells isolated from fetal porcine skin have the potential to form oocyte-like cells (OLCs) in vitro. However, primordial germ cells (PGCs), which must also be specified during the stem cell differentiation to give rise to these putative oocytes at more advanced stages of culture, were not systematically characterized. The current study tested the hypothesis that a morphologically distinct population of cells derived from skin stem cells prior to OLC formation corresponds to putative PGCs, which differentiate further into more mature gametes. METHODOLOGY/PRINCIPAL FINDINGS: When induced to differentiate in an appropriate microenvironment, a subpopulation of morphologically distinct cells, some of which are alkaline phosphatase (AP) positive, also express Oct4, Fragilis, Stella, Dazl, and Vasa, which are markers indicative of germ cell formation. A known differentially methylated region (DMR) within the H19 gene locus, which is demethylated in oocytes after establishment of the maternal imprint, is hypomethylated in PGC-like cells compared to undifferentiated skin-derived stem cells, suggesting that the putative germ cell population undergoes imprint erasure. Additional evidence supporting the germ cell identity of in vitro-generated PGC-like cells is that, when labeled with a Dazl-GFP reporter, these cells further differentiate into GFP-positive OLCs. SIGNIFICANCE: The ability to generate germ cell precursors from somatic stem cells may provide an in vitro model to study some of the unanswered questions surrounding early germ cell formation. PMID- 20011594 TI - A high-throughput DNA sequence aligner for microbial ecology studies. AB - As the scope of microbial surveys expands with the parallel growth in sequencing capacity, a significant bottleneck in data analysis is the ability to generate a biologically meaningful multiple sequence alignment. The most commonly used aligners have varying alignment quality and speed, tend to depend on a specific reference alignment, or lack a complete description of the underlying algorithm. The purpose of this study was to create and validate an aligner with the goal of quickly generating a high quality alignment and having the flexibility to use any reference alignment. Using the simple nearest alignment space termination algorithm, the resulting aligner operates in linear time, requires a small memory footprint, and generates a high quality alignment. In addition, the alignments generated for variable regions were of as high a quality as the alignment of full length sequences. As implemented, the method was able to align 18 full-length 16S rRNA gene sequences and 58 V2 region sequences per second to the 50,000-column SILVA reference alignment. Most importantly, the resulting alignments were of a quality equal to SILVA-generated alignments. The aligner described in this study will enable scientists to rapidly generate robust multiple sequences alignments that are implicitly based upon the predicted secondary structure of the 16S rRNA molecule. Furthermore, because the implementation is not connected to a specific database it is easy to generalize the method to reference alignments for any DNA sequence. PMID- 20011595 TI - Consistent paternity skew through ontogeny in Peron's tree frog (Litoria peronii). AB - BACKGROUND: A large number of studies in postcopulatory sexual selection use paternity success as a proxy for fertilization success. However, selective mortality during embryonic development can lead to skews in paternity in situations of polyandry and sperm competition. Thus, when assessment of paternity fails to incorporate mortality skews during early ontogeny, this may interfere with correct interpretation of results and subsequent evolutionary inference. In a previous series of in vitro sperm competition experiments with amphibians (Litoria peronii), we showed skewed paternity patterns towards males more genetically similar to the female. METHODOLOGY/PRINCIPAL FINDINGS: Here we use in vitro fertilizations and sperm competition trials to test if this pattern of paternity of fully developed tadpoles reflects patterns of paternity at fertilization and if paternity skews changes during embryonic development. We show that there is no selective mortality through ontogeny and that patterns of paternity of hatched tadpoles reflects success of competing males in sperm competition at fertilization. CONCLUSIONS/SIGNIFICANCE: While this study shows that previous inferences of fertilization success from paternity data are valid for this species, rigorous testing of these assumptions is required to ensure that differential embryonic mortality does not confound estimations of true fertilization success. PMID- 20011596 TI - Sexually transmitted infections among HIV-1-discordant couples. AB - INTRODUCTION: More new HIV-1 infections occur within stable HIV-1-discordant couples than in any other group in Africa, and sexually transmitted infections (STIs) may increase transmission risk among discordant couples, accounting for a large proportion of new HIV-1 infections. Understanding correlates of STIs among discordant couples will aid in optimizing interventions to prevent HIV-1 transmission in these couples. METHODS: HIV-1-discordant couples in which HIV-1 infected partners were HSV-2-seropositive were tested for syphilis, chlamydia, gonorrhea, and trichomoniasis, and HIV-1-uninfected partners were tested for HSV 2. We assessed sociodemographic, behavioral, and biological correlates of a current STI. RESULTS: Of 416 couples enrolled, 16% were affected by a treatable STI, and among these both partners were infected in 17% of couples. A treatable STI was found in 46 (11%) females and 30 (7%) males. The most prevalent infections were trichomoniasis (5.9%) and syphilis (2.6%). Participants were 5.9 fold more likely to have an STI if their partner had an STI (P<0.01), and STIs were more common among those reporting any unprotected sex (OR = 2.43; P<0.01) and those with low education (OR = 3.00; P<0.01). Among HIV-1-uninfected participants with an HSV-2-seropositive partner, females were significantly more likely to be HSV-2-seropositive than males (78% versus 50%, P<0.01). CONCLUSIONS: Treatable STIs were common among HIV-1-discordant couples and the majority of couples affected by an STI were discordant for the STI, with relatively high HSV 2 discordance. Awareness of STI correlates and treatment of both partners may reduce HIV-1 transmission. TRIAL REGISTRATION: ClinicalTrials.gov NCT00194519. PMID- 20011597 TI - Kinetic characterization and phosphoregulation of the Francisella tularensis 1 deoxy-D-xylulose 5-phosphate reductoisomerase (MEP synthase). AB - Deliberate and natural outbreaks of infectious disease underscore the necessity of effective vaccines and antimicrobial/antiviral therapeutics. The prevalence of antibiotic resistant strains and the ease by which antibiotic resistant bacteria can be intentionally engineered further highlights the need for continued development of novel antibiotics against new bacterial targets. Isoprenes are a class of molecules fundamentally involved in a variety of crucial biological functions. Mammalian cells utilize the mevalonic acid pathway for isoprene biosynthesis, whereas many bacteria utilize the methylerythritol phosphate (MEP) pathway, making the latter an attractive target for antibiotic development. In this report we describe the cloning and characterization of Francisella tularensis MEP synthase, a MEP pathway enzyme and potential target for antibiotic development. In vitro growth-inhibition assays using fosmidomycin, an inhibitor of MEP synthase, illustrates the effectiveness of MEP pathway inhibition with F. tularensis. To facilitate drug development, F. tularensis MEP synthase was cloned, expressed, purified, and characterized. Enzyme assays produced apparent kinetic constants (K(M)(DXP) = 104 microM, K(M)(NADPH) = 13 microM, k(cat)(DXP) = 2 s(-1), k(cat)(NADPH) = 1.3 s(-1)), an IC(50) for fosmidomycin of 247 nM, and a K(i) for fosmidomycin of 99 nM. The enzyme exhibits a preference for Mg(+2) as a divalent cation. Titanium dioxide chromatography-tandem mass spectrometry identified Ser177 as a site of phosphorylation. S177D and S177E site-directed mutants are inactive, suggesting a mechanism for post-translational control of metabolic flux through the F. tularensis MEP pathway. Overall, our study suggests that MEP synthase is an excellent target for the development of novel antibiotics against F. tularensis. PMID- 20011598 TI - The danger signal adenosine induces persistence of chlamydial infection through stimulation of A2b receptors. AB - Infections with intracellular bacteria such as chlamydiae affect the majority of the world population. Infected tissue inflammation and granuloma formation help contain the short-term expansion of the invading pathogen, leading also to local tissue damage and hypoxia. However, the effects of key aspects of damaged inflamed tissues and hypoxia on continued infection with intracellular bacteria remain unknown. We find that development of Chlamydia trachomatis is reversibly retarded by prolonged exposure of infected cells to extracellular adenosine, a hallmark of hypoxia and advanced inflammation. In epithelial cells, this effect was mediated by the A2b adenosine receptor, unique in the adenosine receptor family for having a hypoxia-inducible factor (HIF1-alpha) binding site at its promoter region, and was dependent on an increase in the intracellular cAMP levels, but was independent of cAMP-dependent protein kinase (PKA). Further study of adenosine receptor signaling during intracellular bacterial infection could lead to breakthroughs in our understanding of persistent infections with these ubiquitous pathogens. PMID- 20011599 TI - Cytoplasmic acidification and the benzoate transcriptome in Bacillus subtilis. AB - BACKGROUND: Bacillus subtilis encounters a wide range of environmental pH. The bacteria maintain cytoplasmic pH within a narrow range. Response to acid stress is a poorly understood function of external pH and of permeant acids that conduct protons into the cytoplasm. METHODS AND PRINCIPAL FINDINGS: Cytoplasmic acidification and the benzoate transcriptome were observed in Bacillus subtilis. Cytoplasmic pH was measured with 4-s time resolution using GFPmut3b fluorimetry. Rapid external acidification (pH 7.5 to 6.0) acidified the B. subtilis cytoplasm, followed by partial recovery. Benzoate addition up to 60 mM at external pH 7 depressed cytoplasmic pH but left a transmembrane Delta pH permitting growth; this robust adaptation to benzoate exceeds that seen in E. coli. Cytoplasmic pH was depressed by 0.3 units during growth with 30 mM benzoate. The transcriptome of benzoate-adapted cells was determined by comparing 4,095 gene expression indices following growth at pH 7, +/- 30 mM benzoate. 164 ORFs showed > or = 2 fold up-regulation by benzoate (30 mM benzoate/0 mM), and 102 ORFs showed > or = 2-fold down-regulation. 42% of benzoate-dependent genes are regulated up or down, respectively, at pH 6 versus pH 7; they are candidates for cytoplasmic pH response. Acid-stress genes up-regulated by benzoate included drug resistance genes (yhbI, yhcA, yuxJ, ywoGH); an oligopeptide transporter (opp); glycine catabolism (gcvPA-PB); acetate degradation (acsA); dehydrogenases (ald, fdhD, serA, yrhEFG, yjgCD); the TCA cycle (citZ, icd, mdh, sucD); and oxidative stress (OYE-family yqjM, ohrB). Base-stress genes down-regulated by benzoate included malate metabolism (maeN), sporulation control (spo0M, spo0E), and the SigW alkali shock regulon. Cytoplasmic pH could mediate alkali-shock induction of SigW. CONCLUSIONS: B. subtilis maintains partial pH homeostasis during growth, and withstands high concentrations of permeant acid stress, higher than for gram negative neutralophile E. coli. The benzoate adaptation transcriptome substantially overlaps that of external acid, contributing to a cytoplasmic pH transcriptome. PMID- 20011600 TI - IgG autoantibody to brain beta tubulin III associated with cytokine cluster-II discriminate cerebral malaria in central India. AB - BACKGROUND: The main processes in the pathogenesis of cerebral malaria caused by Plasmodium falciparum involved sequestration of parasitized red blood cells and immunopathological responses. Among immune factors, IgG autoantibodies to brain antigens are increased in P. falciparum infected patients and correlate with disease severity in African children. Nevertheless, their role in the pathophysiology of cerebral malaria (CM) is not fully defined. We extended our analysis to an Indian population with genetic backgrounds and endemic and environmental status different from Africa to determine if these autoantibodies could be either a biomarker or a risk factor of developing CM. METHODS/PRINCIPAL FINDINGS: We investigated the significance of these self-reactive antibodies in clinically well-defined groups of P. falciparum infected patients manifesting mild malaria (MM), severe non-cerebral malaria (SM), or cerebral malaria (CM) and in control subjects from Gondia, a malaria epidemic site in central India using quantitative immunoprinting and multivariate statistical analyses. A two-fold complete-linkage hierarchical clustering allows classifying the different patient groups and to distinguish the CM from the others on the basis of their profile of IgG reactivity to brain proteins defined by PANAMA Blot. We identified beta tubulin III (TBB3) as a novel discriminant brain antigen in the prevalence of CM. In addition, circulating IgG from CM patients highly react with recombinant TBB3. Overall, correspondence analyses based on singular value decomposition show a strong correlation between IgG anti-TBB3 and elevated concentration of cluster-II cytokine (IFNgamma, IL1beta, TNFalpha, TGFbeta) previously demonstrated to be a predictor of CM in the same population. CONCLUSIONS/SIGNIFICANCE: Collectively, these findings validate the relationship between antibody response to brain induced by P. falciparum infection and plasma cytokine patterns with clinical outcome of malaria. They also provide significant insight into the immune mechanisms associated to CM by the identification of TBB3 as a new disease specific marker and potential therapeutic target. PMID- 20011601 TI - VEGF gene expression in adult human thymus fat: a correlative study with hypoxic induced factor and cyclooxygenase-2. AB - It is well known that the adult human thymus degenerates into fat tissue; however, it has never been considered as a potential source of angiogenic factors. Recently, we have described that this fat (TAT) produces angiogenic factors and induces human endothelial cell proliferation and migration, indicating its potential angiogenic properties. DESIGN: Adult thymus fat and subcutaneous adipose tissue specimens were obtained from 28 patients undergoing cardiac surgery, making this tissue readily available as a prime source of adipose tissue. We focused our investigation on determining VEGF gene expression and characterizing the different genes, mediators of inflammation and adipogenesis, and which are known to play a relevant role in angiogenesis regulation. RESULTS: We found that VEGF-A was the isoform most expressed in TAT. This expression was accompanied by an upregulation of HIF-1alpha, COX-2 and HO-1 proteins, and by increased HIF-1 DNA binding activity, compared to SAT. Furthermore, we observed that TAT contains a high percentage of mature adipocytes, 0.25% of macrophage cells, 15% of endothelial cells and a very low percentage of thymocyte cells, suggesting the cellular variability of TAT, which could explain the differences in gene expression observed in TAT. Subsequently, we showed that the expression of genes known as adipogenic mediators, including PPARgamma1/gamma2, FABP-4 and adiponectin was similar in both TAT and SAT. Moreover the expression of these latter genes presented a significantly positive correlation with VEGF, suggesting the potential association between VEGF and the generation of adipose tissue in adult thymus. CONCLUSION: Here we suggest that this fat has a potential angiogenic function related to ongoing adipogenesis, which substitutes immune functions within the adult thymus. The expression of VEGF seems to be associated with COX-2, HO-1 and adipogenesis related genes, suggesting the importance that this new fat has acquired in research in relation to adipogenesis and angiogenesis. PMID- 20011603 TI - High and far: biases in the location of protected areas. AB - BACKGROUND: About an eighth of the earth's land surface is in protected areas (hereafter "PAs"), most created during the 20(th) century. Natural landscapes are critical for species persistence and PAs can play a major role in conservation and in climate policy. Such contributions may be harder than expected to implement if new PAs are constrained to the same kinds of locations that PAs currently occupy. METHODOLOGY/PRINCIPAL FINDINGS: Quantitatively extending the perception that PAs occupy "rock and ice", we show that across 147 nations PA networks are biased towards places that are unlikely to face land conversion pressures even in the absence of protection. We test each country's PA network for bias in elevation, slope, distances to roads and cities, and suitability for agriculture. Further, within each country's set of PAs, we also ask if the level of protection is biased in these ways. We find that the significant majority of national PA networks are biased to higher elevations, steeper slopes and greater distances to roads and cities. Also, within a country, PAs with higher protection status are more biased than are the PAs with lower protection statuses. CONCLUSIONS/SIGNIFICANCE: In sum, PAs are biased towards where they can least prevent land conversion (even if they offer perfect protection). These globally comprehensive results extend findings from nation-level analyses. They imply that siting rules such as the Convention on Biological Diversity's 2010 Target [to protect 10% of all ecoregions] might raise PA impacts if applied at the country level. In light of the potential for global carbon-based payments for avoided deforestation or REDD, these results suggest that attention to threat could improve outcomes from the creation and management of PAs. PMID- 20011602 TI - Angiotensin I-converting enzyme mutation (Trp1197Stop) causes a dramatic increase in blood ACE. AB - BACKGROUND: Angiotensin-converting enzyme (ACE) metabolizes many peptides and plays a key role in blood pressure regulation and vascular remodeling. Elevated ACE levels may be associated with an increased risk for different cardiovascular or respiratory diseases, including asthma. Previously, a molecular mechanism underlying a 5-fold familial increase of blood ACE was discovered: Pro1199Leu substitution enhanced the cleavage-secretion process. Carriers of this mutation were Caucasians from Europe (mostly Dutch) or had European roots. METHODOLOGY/PRINCIPAL FINDINGS: We have found a family of African-American descent whose affected members' blood ACE level was increased 13-fold over normal. In affected family members, codon TGG coding for Trp1197 was substituted in one allele by TGA (stop codon). As a result, half of ACE expressed in these individuals had a length of 1196 amino acids and lacked a transmembrane anchor. This ACE mutant is not trafficked to the cell membrane and is directly secreted out of cells; this mechanism apparently accounts for the high serum ACE level seen in affected individuals. A haplotype of the mutant ACE allele was determined based on 12 polymorphisms, which may help to identify other carriers of this mutation. Some but not all carriers of this mutation demonstrated airflow obstruction, and some but not all have hypertension. CONCLUSIONS/SIGNIFICANCE: We have identified a novel Trp1197Stop mutation that results in dramatic elevation of serum ACE. Since blood ACE elevation is often taken as a marker of disease activity (sarcoidosis and Gaucher diseases), it is important for clinicians and medical scientists to be aware of alternative genetic causes of elevated blood ACE that are not apparently linked to disease. PMID- 20011604 TI - A phosphoinositide 3-kinase/phospholipase Cgamma1 pathway regulates fibroblast growth factor-induced capillary tube formation. AB - BACKGROUND: The fibroblast growth factors (FGFs) are key regulators of embryonic development, tissue homeostasis and tumour angiogenesis. Binding of FGFs to their receptor(s) results in activation of several intracellular signalling cascades including phosphoinositide 3-kinase (PI3K) and phospholipase C (PLC)gamma1. Here we investigated the basic FGF (FGF-2)-mediated activation of these enzymes in human umbilical vein endothelial cells (HUVECs) and defined their role in FGF-2 dependent cellular functions. METHODOLOGY/PRINCIPAL FINDINGS: We show that FGF-2 activates PLCgamma1 in HUVECs measured by analysis of total inositol phosphates production upon metabolic labelling of cells and intracellular calcium increase. We further demonstrate that FGF-2 activates PI3K, assessed by analysing accumulation of its lipid product phosphatidylinositol-3,4,5-P(3) using TLC and confocal microscopy analysis. PI3K activity is required for FGF-2-induced PLCgamma1 activation and the PI3K/PLCgamma1 pathway is involved in FGF-2 dependent cell migration, determined using Transwell assay, and in FGF-2-induced capillary tube formation (tubulogenesis assays in vitro). Finally we show that PI3K-dependent PLCgamma1 activation regulates FGF-2-mediated phosphorylation of Akt at its residue Ser473, determined by Western blotting analysis. This occurs through protein kinase C (PKC)alpha activation since dowregulation of PKCalpha expression using specific siRNA or blockade of its activity using chemical inhibition affects the FGF-2-dependent Ser473 Akt phosphorylation. Furthermore inhibition of PKCalpha blocks FGF-2-dependent cell migration. CONCLUSION/SIGNIFICANCE: These data elucidate the role of PLCgamma1 in FGF-2 signalling in HUVECs demonstrating its key role in FGF-2-dependent tubulogenesis. Furthermore these data unveil a novel role for PLCgamma1 as a mediator of PI3K dependent Akt activation and as a novel key regulator of different Akt-dependent processes. PMID- 20011605 TI - Gene structure induced epigenetic modifications of pericarp color1 alleles of maize result in tissue-specific mosaicism. AB - BACKGROUND: The pericarp color1 (p1) gene encodes for a myb-homologous protein that regulates the biosynthesis of brick-red flavonoid pigments called phlobahpenes. The pattern of pigmentation on the pericarp and cob glumes depends upon the allelic constitution at the p1 locus. p1 alleles have unique gene structure and copy number which have been proposed to influence the epigenetic regulation of tissue-specific gene expression. For example, the presence of tandem-repeats has been correlated with the suppression of pericarp pigmentation though a mechanism associated with increased DNA methylation. METHODOLOGY/PRINCIPAL FINDINGS: Herein, we extensively characterize a p1 allele called P1-mosaic (P1-mm) that has mosaic pericarp and light pink or colorless cob glumes pigmentation. Relative to the P1-wr (white pericarp and red cob glumes), we show that the tandem repeats of P1-mm have a modified gene structure containing a reduced number of repeats. The P1-mm has reduced DNA methylation at a distal enhancer and elevated DNA methylation downstream of the transcription start site. CONCLUSIONS/SIGNIFICANCE: Mosaic gene expression occurs in many eukaryotes. Herein we use maize p1 gene as model system to provide further insight about the mechanisms that govern expression mosaicism. We suggest that the gene structure of P1-mm is modified in some of its tandem gene repeats. It is known that repeated genes are susceptible to chromatin-mediated regulation of gene expression. We discuss how the modification to the tandem repeats of P1-mm may have disrupted the epigenetic mechanisms that stably confer tissue-specific expression. PMID- 20011606 TI - Chaperonin contributes to cold hardiness of the onion maggot Delia antiqua through repression of depolymerization of actin at low temperatures. AB - Winter-diapause and cold-acclimated non-diapause pupae of the onion maggot, Delia antiqua (Diptera: Anthomyiidae), show strong cold hardiness. To obtain insights into the mechanisms involved in the enhancement of cold hardiness, we investigated the expression patterns of genes encoding subunits of chaperonin (CCT) and the morphology of actin, a substrate of CCT, at low temperatures. Quantitative real-time PCR analyses showed the mRNA levels of CCT subunits in pupal tissues to be highly correlated with the cold hardiness of the pupae. While actin in the Malpighian tubules of non-cold-hardy pupae showed extensive depolymerization after a cold treatment, actin in the same tissue of cold-hardy pupae was not depolymerized. Damage to cell membranes became apparent after the depolymerization of actin. Moreover, administration of Latrunculin B, an inhibitor of actin polymerization, to the larvae markedly decreased the cold hardiness of the pupae obtained. These findings suggest that CCT contributes to the cold hardiness of D. antiqua through the repression of depolymerization of actin at low temperatures. PMID- 20011607 TI - Activation and maintenance of peripheral semantic features of unambiguous words after right hemisphere brain damage in adults. AB - BACKGROUND: The right cerebral hemisphere (RH) sustains activation of subordinate, secondary, less common, and/or distantly related meanings of words. Much of the pertinent data come from studies of homonyms, but some evidence also suggests that the RH has a unique maintenance function in relation to unambiguous nouns. In a divided visual field priming study, Atchley, Burgess, and Keeney (1999) reported that only left visual field/RH presentation yielded evidence of continuing activation of peripheral semantic features that were incompatible with the most common image or representation of their corresponding nouns (e.g., rotten for "apple"). Activation for weakly related features that were compatible with the dominant representation (e.g., crunchy) was sustained over time regardless of the visual field/hemisphere of initial stimulus input. Several studies report that unilateral right hemisphere brain damage (RHD) in adults affects the RH's meaning maintenance function, but this work also has centred on homonyms, and/or more recently metonymic and metaphoric polysemous words. AIMS: The current investigation examined whether RHD deficits in processing secondary and/or distantly related meanings of words, typically observed in studies of homonyms, would extend to peripheral, weakly related semantic features of unambiguous nouns. METHODS #ENTITYSTARTX00026; PROCEDURES: Participants were 28 adults with unilateral RHD from cerebrovascular accident, and 38 adults without brain damage. Participants listened to spoken sentences that ended with an unambiguous noun. Each sentence was followed by a spoken target phoneme string. Targets included peripheral semantic features of the sentence-final noun that were either compatible or incompatible with the dominant mental images of the noun, and were presented at two intervals after that noun. A lexical decision task was used to gauge both the early activation and maintenance of activation for these weakly related semantic features. OUTCOMES #ENTITYSTARTX00026; RESULTS: Accuracy data demonstrated activation (priming) for both types of peripheral features, in both groups, shortly after presentation of the corresponding noun. Neither group evidenced continuing activation for either type of feature at a longer interval. These results are interpreted as reflecting rapid decay/poor maintenance of activation for distantly related features for both groups. The lack of a biasing context, however, did not provide an appropriate test for previously reported suppression deficits after RHD. Fast decay of activation of compatible semantic features was unexpected for the control group. Adults with RHD were less accurate than the control group at both test intervals for the features that are semantically more distant from their associated nouns (Related-incompatible features). Accordingly, it is argued that the RHD group's poor maintenance of activation for these features reflects a deficit, rather than normal performance. The interpretation of results from this study is complicated by the lack of RT priming for either type of semantic feature, and for either participant group. CONCLUSIONS: The right cerebral hemisphere appears to be necessary for activating semantic features that are particularly distantly related to their corresponding lexical items, and for sustaining activation of these features in the absence of a biasing context. Because lexical processing has been linked with discourse comprehension for adults with RHD, more work in this area should enhance clinical management in the future. PMID- 20011608 TI - The Link between the Marital Bond and Future Triadic Family Interactions. AB - This study examined how the marital bond, as indexed through the Oral History Interview (OHI), is related to future triadic family interactions. Families (N = 108), with a 7 - 9 year old child, participated in a longitudinal study (the Family Health Project) examining children's emotional development throughout the transition to adolescence. Parental cohesion and family cohesion, warmth, structure, and problem solving were assessed via behavioral observation during family problem solving discussions and parent-child teaching interactions 18 - 24 months after the OHI. Results indicated that the marital bond was predictive of parental cohesion, family cohesion, warmth, and structure during teaching interactions. The marital bond was not significantly predictive of family problem solving or parental cohesion in problem solving interactions. PMID- 20011610 TI - m-Aminophenylboronic Acid as a Functional Monomer for Fabricating Molecularly Imprinted Polymer for the Recognition of Bovine Serum Albumin. AB - Using m-aminophenylboronic acid (APBA) as a functional monomer, molecularly imprinted polymer (MIP) imprinted with bovine serum albumin (BSA) was fabricated on activated glass spheres under optimized conditions. Key factors in the prepolymerization reaction (between APBA and BSA), such as buffer pH, ionic strength and reaction time, were carefully optimized as previously reported [1]. The interaction between APBA and BSA during the prepolymerization stage was investigated and optimized, and ideal conditions for protein rebinding experiments were determined as well. Protein rebinding and enriching properties of polymers were studied in single and competitive binding protocols, respectively. The key point of the present paper is that the binding selectivity of polymers may be estimated by the amount of bound-protein recovered from a protein-saturated polymer. Results demonstrated that the selectivity of MIP towards its template protein is superior to that of non-imprinted polymer (NIP). PMID- 20011611 TI - Influenza vaccination in the elderly: seeking new correlates of protection and improved vaccines. AB - Influenza is foremost among all infectious diseases for an age-related increase in risk for serious complications and death. Determining the benefit of current influenza vaccines is largely limited to epidemiologic studies, since placebo controlled trials of influenza vaccines are no longer considered ethical in the older adult population. Vaccine effectiveness is calculated from the relative reduction in influenza outcomes in individuals who elect to be vaccinated compared with those who do not, the assumptions for which are diverse and have led to considerable controversy as to the exact benefit of influenza vaccination in older adults. In spite of this controversy, there is no doubt that new influenza vaccine technologies are needed to improve protection and reverse the trend of rising hospitalization and death rates related to influenza in older adults despite widespread influenza vaccination programs. This article will review the challenges to new vaccine development, explore the potential correlates of protection against influenza, and describe how new vaccine technologies may improve protection against complicated influenza illness in the older adult population. PMID- 20011612 TI - Nominations for Sale. AB - Models of nomination politics in the US often find "gridlock" in equilibrium because of the super-majority requirement in the Senate for the confirmation of presidential nominees. A blocking coalition often prefers to defeat any nominee. Yet empirically nominations are successful. In the present paper we explore the possibility that senators can be induced to vote contrary to their nominal (gridlock-producing) preferences through contributions from the president and/or lobbyists, thus breaking the gridlock and confirming the nominee. We model contributions by the president and lobbyists according to whether payment schedules are conditioned on the entire voting profile, the vote of a senator, or the outcome. We analyze several extensions to our baseline approach, including the possibility that lobbyists may find it more productive to offer inducements to the president in order to affect his proposal behavior, rather than trying to induce senators to vote for or against a given nominee. PMID- 20011613 TI - Computational Systems Bioinformatics and Bioimaging for Pathway Analysis and Drug Screening. AB - The premise of today's drug development is that the mechanism of a disease is highly dependent upon underlying signaling and cellular pathways. Such pathways are often composed of complexes of physically interacting genes, proteins, or biochemical activities coordinated by metabolic intermediates, ions, and other small solutes and are investigated with molecular biology approaches in genomics, proteomics, and metabonomics. Nevertheless, the recent declines in the pharmaceutical industry's revenues indicate such approaches alone may not be adequate in creating successful new drugs. Our observation is that combining methods of genomics, proteomics, and metabonomics with techniques of bioimaging will systematically provide powerful means to decode or better understand molecular interactions and pathways that lead to disease and potentially generate new insights and indications for drug targets. The former methods provide the profiles of genes, proteins, and metabolites, whereas the latter techniques generate objective, quantitative phenotypes correlating to the molecular profiles and interactions. In this paper, we describe pathway reconstruction and target validation based on the proposed systems biologic approach and show selected application examples for pathway analysis and drug screening. PMID- 20011614 TI - Extensions of the Ferry shear wave model for active linear and nonlinear microrheology. AB - The classical oscillatory shear wave model of Ferry et al. [J. Polym. Sci. 2:593 611, (1947)] is extended for active linear and nonlinear microrheology. In the Ferry protocol, oscillation and attenuation lengths of the shear wave measured from strobe photographs determine storage and loss moduli at each frequency of plate oscillation. The microliter volumes typical in biology require modifications of experimental method and theory. Microbead tracking replaces strobe photographs. Reflection from the top boundary yields counterpropagating modes which are modeled here for linear and nonlinear viscoelastic constitutive laws. Furthermore, bulk imposed strain is easily controlled, and we explore the onset of normal stress generation and shear thinning using nonlinear viscoelastic models. For this paper, we present the theory, exact linear and nonlinear solutions where possible, and simulation tools more generally. We then illustrate errors in inverse characterization by application of the Ferry formulas, due to both suppression of wave reflection and nonlinearity, even if there were no experimental error. This shear wave method presents an active and nonlinear analog of the two-point microrheology of Crocker et al. [Phys. Rev. Lett. 85: 888 - 891 (2000)]. Nonlocal (spatially extended) deformations and stresses are propagated through a small volume sample, on wavelengths long relative to bead size. The setup is ideal for exploration of nonlinear threshold behavior. PMID- 20011615 TI - The Nature-Nurture Debate and Public Policy. AB - The contentious nature-nurture debate in developmental psychology is poised to reach a rapprochement with contemporary concepts of gene-environment interaction, transaction, and fit. Discoveries over the past decade have revealed how neither genes nor the environment offers a sufficient window into human development. Rather, the most important discoveries have come from unearthing the manner in which the environment alters gene expression (and how genes impose limits on environmental effects), how biology and the environment influence each other across time, and how maximizing gene-environment fit leads to optimal outcomes for children. The manner in which these factors operate in tandem should direct future scholarship, practice, and public policy. PMID- 20011616 TI - Smoking and Cancers: Case-robust Analysis of a Classic Data Set. AB - A typical structural equation model is intended to reproduce the means, variances, and correlations or covariances among a set of variables based on parameter estimates of a highly restricted model. It is not widely appreciated that the sample statistics being modeled can be quite sensitive to outliers and influential observations leading to bias in model parameter estimates. A classic public epidemiological data set on the relation between cigarette purchases and rates of four types of cancer among states in the USA is studied with case weighting methods that reduce the influence of a few cases on the overall results. The results support and extend the original conclusions; the standardized effect of smoking on a factor underlying deaths from bladder and lung cancer is .79. PMID- 20011617 TI - Rapid Formation of Acrylated Microstructures by Microwave-Induced Thermal Crosslinking. AB - We present a rapid and highly efficient method to form microstructure of poly(ethylene glycol) (PEG)-based acrylates by microwave-induced thermal crosslinking. PEG-based polymeric microstructures such as polymer microarrays and microwells were fabricated on 3-(trimethoxysilyl)propyl methacrylate (TMSPMA) coated glass slides that were placed on top of a silicon wafer. In comparison to ultraviolet (UV) irradiation curing, microwave-induced thermal crosslinking could be completed within 10 s, without thermal degradation or oxygen inhibition in the presence of ambient oxygen. Furthermore, the activation of surviving free radical impurities by microwave-induced heating enabled crosslinking even without an exogenous radical initiator (e.g., 2,2'-azoisobutyronitrile (AIBN)). This approach can be beneficial for fabricating various PEG-based microstructures for high-throughput screening assays, cell-based biosensors, and biomedical microdevices. PMID- 20011618 TI - PET imaging for Treatment Response in Cancer. PMID- 20011619 TI - Dielectrophoretic Separation of Cancer Cells from Blood. AB - Recent measurements have demonstrated that the dielectric properties of cells depend on their type and physiological status. For example, MDA-231 human breast cancer cells were found to have a mean plasma membrane specific capacitance of 26 mF/m(2), more than double the value (11 mF/m(2)) observed for resting T lymphocytes. When an inhomogeneous ac electric field is applied to a particle, a dielectrophoretic (DEP) force arises that depends on the particle dielectric properties. Therefore, cells having different dielectric characteristics will experience differential DEP forces when subjected to such a field. In this article, we demonstrate the use of differential DEP forces for the separation of several different cancerous cell types from blood in a dielectric affinity column. These separations were accomplished using thin, flat chambers having microelectrode arrays on the bottom wall. DEP forces generated by the application of ac fields to the electrodes were used to influence the rate of elution of cells from the chamber by hydrodynamic forces within a parabolic fluid flow profile. Electrorotation measurements were first made on the various cell types found within cell mixtures to be separated, and theoretical modeling was used to derive the cell dielectric parameters. Optimum separation conditions were then predicted from the frequency and suspension conductivity dependencies of cell DEP responses defined by these parameters. Cell separations were then undertaken for various ratios of cancerous to normal cells at different concentrations. Eluted cells were characterized in terms of separation efficiency, cell viability, and separation speed. For example, 100% efficiency was achieved for purging MDA-231 cells from blood at the tumor to normal cell ratio 1:1 x 10(5) or 1:3 x 10(5), cell viability was not compromised, and separation rates were at least 10(3) cells/s. Theoretical and experimental criteria for the design and operation of such separators are presented. PMID- 20011620 TI - Enantioselective Ag-Catalyzed Allylation of Aldimines. AB - A highly enantioselective synthesis of homoallylic amines, using allyltrimethoxysilane under Ag(I) catalytic conditions, has been developed. Among the chiral ligands investigated, a remarkable difference in the resulting Ag(I) complexes was observed. Under mild conditions and low catalyst loadings, homoallylamines were produced in high ee values (up to 80%) and good yields. The methodology can be further extended to a diastere- and enantioselective crotylation of aldimines. PMID- 20011621 TI - PARENTAL DEATH IN THE LIVES OF PEOPLE WITH SERIOUS MENTAL ILLNESS. AB - Nearly a fourth (22%) of the participants within a research sample of 148 individuals with serious mental illness reported the death of a loved one as a significant loss, and two thirds of these deaths involved the loss of one or both parents. The key determinant of the severity and duration of grief in response to the death of a parent was whether or not there were extenuating circumstances that complicated the death event, such as co-residence with the deceased at the time of death or a lack of regular social contact with anyone other than the deceased. In all instances of severe or prolonged grief, there was no preparation for the parental death, either through preparatory counseling or practical plans for funeral arrangements, financial repercussions, life-style changes, or residential relocation. Mental health agencies serving people with serious mental illness should begin to incorporate financial and emotional preparation for parental deaths and bereavement counseling as essential services. PMID- 20011622 TI - Declarative Memory. AB - Neuroimaging of declarative memory is not an endeavor divorced from psychology but, instead, is another path through which a more complete understanding of memory has emerged. Specifically, neuroimaging allows us to determine if differences between memory states emerge from quantitatively or qualitatively distinct underlying encoding operations. Further, it has allowed for greater specification of the putative control operations adopted when we make decisions about our memories. We describe some examples of insights provided by neuroimaging into the many and varied processes that support encoding and retrieval of declarative memory. PMID- 20011623 TI - Shear Stress Regulates the Flk-1/Cbl/PI3K/NF-kappaB Pathway Via Actin and Tyrosine Kinases. AB - Vascular endothelial cells (ECs) are continuously exposed to mechanical stimuli (e.g., shear stress). Our previous study has shown that the shear-induced nuclear factor-kappaB (NF-kappaB) activation is mediated by integrins [Bhullar, I. S., Y. S. Li, H. Miao, E. Zandi, M. Kim, et al. J. Biol. Chem. 273:30544-30549, 1998]. The shear-activated integrins can also transactivate Flk-1 (a receptor for vascular endothelial growth factor (VEGF)) [Wang, Y., H. Miao, S. Li, K. D. Chen, Y. S. Li, et al. Am. J. Physiol. Cell Physiol. 283:C1540-C1547, 2002], which subsequently recruits Casitas B-lineage lymphoma (Cbl) to regulate inhibitor of kappaB protein kinase (IKK) [Wang, Y., J. Chang, Y. C. Li, Y. S. Li, J. Y. Shyy, and S. Chien. Am. J. Physiol. Heart Circ. Physiol. 286:H685-H692, 2004], an upstream molecule of NF-kappaB. Therefore, shear stress may likely utilize the Flk-1/Cbl pathway in regulating NF-kappaB. In this paper, we confirmed that the inhibition of Flk-1 by its specific inhibitor SU1498 blocked the shear-induced NF kappaB translocation. The inhibition of Cbl (an adaptor protein which binds to Flk-1 upon shear) by using a negative mutant (Cbl(nm)) also blocked the promoter activity of NF-kappaB, and the inhibition of the Cbl-downstream molecule phosphatidylinositol-3-kinase (PI3K) abolished the NF-kappaB translocation. Further experiments revealed that the disruption of actin cytoskeleton inhibited the Flk-1 and Cbl interaction and NF-kappaB translocation. The inhibition of focal adhesion kinase (FAK) and Src family kinases, which are involved in the integrin-mediated focal adhesion complex, also blocked the shear-induced NF kappaB translocation. Together with our previous findings that integrins mediate the shear-induced activation of Flk-1 and NF-kappaB [Bhullar, I. S., Y. S. Li, H. Miao, E. Zandi, M. Kim, et al. J. Biol. Chem. 273:30544-30549, 1998; Wang, Y., H. Miao, S. Li, K. D. Chen, Y. S. Li, et al. Am. J. Physiol. Cell Physiol. 283:C1540 C1547, 2002], the present results suggest that Flk-1, Cbl, and PI3K act upstream to NF-kappaB in response to shear stress. This Flk-1/Cbl/PI3K/NF-kappaB signaling pathway may be originated from integrins and transmitted by key tyrosine kinases and actin cytoskeleton. These results shed new lights on the molecular mechanism by which mechanical shear stress activates the NF-kappaB signaling pathway, which is critical for vascular inflammatory responses and atherosclerosis. PMID- 20011624 TI - Bayesian Adaptation of the Summary ROC Curve Method for Meta-analysis of Diagnostic Test Performance. AB - Meta-analytic methods for diagnostic test performance, Bayesian methods in particular, have not been well developed. The most commonly used method for meta analysis of diagnostic test performance is the Summary Receiver Operator Characteristic (SROC) curve approach of Moses, Shapiro and Littenberg. In this paper, we provide a brief summary of the SROC method, then present a case study of a Bayesian adaptation of their SROC curve method that retains the simplicity of the original model while additionally incorporating uncertainty in the parameters, and can also easily be extended to incorporate the effect of covariates. We further derive a simple transformation which facilitates prior elicitation from clinicians. The method is applied to two datasets: an assessment of computed tomography for detecting metastases in non-small-cell lung cancer, and a novel dataset to assess the diagnostic performance of endoscopic ultrasound (EUS) in the detection of biliary obstructions relative to the current gold standard of endoscopic retrograde cholangiopancreatography (ERCP). PMID- 20011625 TI - Free Energy Calculations for DNA Near Surfaces Using an Ellipsoidal Geometry. AB - The change in some thermodynamic quantities such as Gibbs' free energy, entropy and enthalpy of the binding of two DNA strands (forming a double helix), while one is tethered to a surface and are analytically calculated. These particles are submerged in an electrolytic solution; the ionic strength of the media allows the linearized version of the Poisson-Boltzmann equation (from the theory of the double layer interaction) to properly describe the interactions [13]. There is experimental and computational evidence that an ion penetrable ellipsoid is an adequate model for the single strand and the double helix [22-25]. The analytic solution provides simple calculations useful for DNA chip design. The predicted electrostatic effects suggest the feasibility of electronic control and detection of DNA hybridization in the fast growing area of DNA recognition. PMID- 20011626 TI - Absolute Single-Molecule Entropies from Quasi-Harmonic Analysis of Microsecond Molecular Dynamics: Correction Terms and Convergence Properties. AB - The convergence properties of the absolute single-molecule configurational entropy and the correction terms used to estimate it are investigated using microsecond molecular dynamics simulation of a peptide test system and an improved methodology. The results are compared with previous applications for systems of diverse chemical nature. It is shown that (i) the effect of anharmonicity is small, (ii) the effect of pairwise correlation is typically large, and (iii) the latter affects to a larger extent the entropy estimate of thermodynamic states characterized by a higher motional correlation. The causes of such deviations from a quasi-harmonic behavior are explained. This improved approach provides entropies also for molecular systems undergoing conformational transitions and characterized by highly frustrated energy surfaces, thus not limited to systems sampling a single quasi-harmonic basin. Overall, this study emphasizes the need for extensive phase-space sampling in order to obtain a reliable estimation of entropic contributions. PMID- 20011627 TI - Influence of sex on gene expression in human corneal epithelial cells. AB - PURPOSE: Sex-associated differences have been identified in the anatomy, physiology and pathophysiology of the human cornea. We hypothesize that many of these differences are due to fundamental variations in gene expression. Our objective in this study was to determine whether such differences exist in human corneal epithelial cells both in vivo and in vitro. METHODS: Human corneal epithelial cells were isolated from the corneoscleral rims of male and female donors. Cells were processed either directly for RNA extraction, or first cultured in phenol red-free keratinocyte serum-free media. The RNA samples were examined for differentially expressed mRNAs by using of CodeLink Bioarrays and Affymetrix GeneChips. Data were analyzed with GeneSifter.Net software. RESULTS: Our results demonstrate that sex significantly influences the expression of over 600 genes in human corneal epithelial cells in vivo. These genes are involved in a broad spectrum of biologic processes, molecular functions and cellular components, such as metabolic processes, DNA replication, cell migration, RNA binding, oxidoreductase activity and nucleoli. We also identified significant, sex-related effects on gene expression in human corneal epithelial cells in vitro. However, with few exceptions (e.g., X- and Y-linked genes), these sex related differences in gene expression in vitro were typically different than those in vivo. CONCLUSIONS: Our findings support our hypothesis that sex-related differences exist in the gene expression of human corneal epithelial cells. Variations in gene expression may contribute to sex-related differences in the prevalence of certain corneal diseases. PMID- 20011628 TI - Prevention of selenite-induced cataractogenesis by rutin in Wistar rats. AB - PURPOSE: To investigate whether rutin retards selenite-induced cataractogenesis in Wistar rat pups. METHODS: On postpartum day ten, Group I rat pups received an intraperitoneal injection of saline. Group II and III rat pups received a subcutaneous injection of sodium selenite. Group III also received an intraperitoneal injection of rutin once daily on postpartum days 9-14. Both eyes of each pup were examined from day 16 up to postpartum day 30. After sacrifice, extricated pup lenses were analyzed for mean activities of catalase, superoxide dismutase, glutathione peroxidase, glutathione S-transferase, and glutathione reductase. In addition, the mean concentrations of reduced glutathione (GSH) and of malondialdehyde were analyzed in samples of lenses and hemolysate. RESULTS: There was dense lenticular opacification in all of Group II, minimal opacification in 33.3% of Group III, no opacification in 66.7% of Group III, and no opacification in Group I. Significantly lower mean activities of lenticular antioxidant enzymes were noted in Group II, compared to Group I and III. Significantly lower mean concentrations of GSH and higher mean concentrations of malondialdehyde were noted in samples of hemolysate and lens from Group II, compared to the values in Group I and III. CONCLUSION: Rutin prevents experimental selenite-induced cataractogenesis in rat pups, possibly by preventing depletion of antioxidant enzymes and of GSH, and by inhibiting lipid peroxidation. PMID- 20011629 TI - The hepatocyte growth factor receptor (MET) gene is not associated with refractive error and ocular biometrics in a Caucasian population. AB - PURPOSE: The purpose of this study was to determine if genetic variants in the hepatocyte growth factor receptor (MET) gene are associated with refractive error and ocular biometric measures in a Caucasian cohort. METHODS: A case-control association study using 818 Caucasian adults (37.2% male, 62.8% female; average age: 51.21+/-17.17 years) was undertaken. All individuals were genotyped for 16 tag single nucleotide polymorphisms (tSNPs) across the MET gene region. Myopia was defined as -0.5 DS or worse in both eyes and divided into high myopia ( 0.001). CONCLUSION: The high seroprevalence of blood-borne pathogens among prospective blood donors in Osogbo, Nigeria calls for mandatory routine screening of blood donors for HBV, HIV, HCV and syphilis. PMID- 20011641 TI - Utility of adsorption techniques in serological evaluation of warm autoimmune haemolytic anaemia. AB - BACKGROUND: Various adsorption techniques are available to remove serum autoantibodies and subsequently detect the underlying alloantibody in previously transfused patients with autoimmune haemolytic anaemia. We planned to establish a suitable adsorption technique in our transfusion service which can remove all autoantibodies and detect underlying alloantibodies rapidly, cheaply and effectively. STUDY DESIGN AND METHODS: We evaluated 71 direct antiglobulin test reactive patients with warm AIHA over a period of 20 months. Twenty-three of these 71 patients who had a previous history of blood transfusion or pregnancy and were confirmed carriers of autoantibodies (indirect antiglobulin test reactive) were considered for the adsorption study. Depending on the adequacy of samples, history of blood transfusion and severity of anaemia either autoadsorption or alloadsorption or both using polyethylene glycol (PEG) or low ionic strength saline (LISS)-papain were performed. RESULTS: Underlying alloantibodies were detected in 7 of the 23 patients (30.4%) and all these were specific to Rhesus antigens. The mean number of alloadsorptions for complete autoantibody removal using PEG was 1.43 which was significantly lower than the 3.9 using the LISS-papain method (p<0.05). The mean time required by PEG alloadsorption and LISS-papain alloadsorption for autoantibody removal was 93.6 minutes and 177.7 minutes, respectively (p<0.05). Discordant results were not observed in any case and identical alloantibodies were detected by both the techniques. CONCLUSION: We found that the PEG method is a rapid, cheap and effective way to remove autoantibodies and detect underlying alloantibodies. PMID- 20011643 TI - Patients' positive identification systems. AB - BACKGROUND: Blood safety must be maintained throughout the whole transfusion chain to prevent the transfusion of incorrect blood components. The estimated risk of an incorrect transfusion is in the order of 1 per 10,000 units of blood. Although several kinds of errors contribute to "wrong blood" events, 70% of errors occur in clinical areas with the most common being due to failure of the pre-transfusion bedside checking procedure. MATERIALS AND METHODS: Several methods are available to reduce such errors. The I-TRAC Plus system by Immucor consists of an identification bracelet which is a bar-coded wristband and a handheld portable computer that identifies patients and blood bags by a scanner and prints the information through a portable printer. The labels attached on the blood order forms and on the sample tubes are read and recorded in the blood bank's informatics system (EmoNet INSIEL). Labels showing the bar-code of the assigned number, which includes the ID number of the patient, the ID number of the unit and a code identifying the kind of product and use (allogeneic or autologous), are generated and applied to the blood components. The transfusions are administered after checking the unit and the patient's wristband using the scanner of a portable PC. RESULTS: In 5 years a total of 71,400 units of blood components were transfused to 15,430 patients using the I-TRAC Plus system. The system prevented 12 cases of mis-identification of patients (5 in 2003, 0 in 2004, 1 in 2005, 1 in 2006 and 5 in 2007). CONCLUSIONS: In 2003 we introduced the use of a bar-code matching system between a patient's wristband and the blood bag to avoid mistakes at the bedside. In 5 years the system provided benefits by avoiding errors in the identification of patients, thus preventing "wrong blood" transfusions. PMID- 20011642 TI - Expression and purification of recombinant human coagulation factor VII fused to a histidine tag using Gateway technology. AB - BACKGROUND: Factor VII (FVII) is a plasma glycoprotein that participates in the coagulation process leading to the generation of fibrin. The aim of this study was to construct, express and purify recombinant FVII fused to a polyhistidine (his) tag using Gateway technology. METHODS: To construct the entry clone, blunt end FVII cDNA and subsequent polymerase chain reaction (PCR) product isolated from a HepG2 cell line was TOPO-cloned into a pENTR TOPO vector. To construct the expression clone, a LR recombination reaction was carried out between the entry clone and destination vector, pDEST26. Chinese hamster ovary (CHO) cells were transfected with 1 microg of DNA of PDEST26-FVII using the FuGENE HD transfection reagent. Two cell lines that permanently expressed recombinant FVII were established. The expression of recombinant FVII was confirmed by reverse transcriptase PCR and enzyme-linked immunosorbent assay. Culture medium containing his-tagged FVII was added to the nickel-nitrilotriacetic acid resin column and bound protein was eluted. The purified protein was detected by sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) and western blot analysis. The biological activity of the recombinant FVII was determined by a prothrombin time assay using FVII-depleted plasma. RESULTS: The results showed that human recombinant FVII was successfully cloned and the accuracy of the nucleotide sequence of the gene and its frame in the vector were confirmed by DNA sequencing. Stable clones transfected with the construct expressed FVII mRNA and related protein but no expression was detected in the CHO cells containing an empty vector. A protein of about 52 KDa was detected in SDS-PAGE and was further confirmed by western blot analysis. A three-fold decrease in clotting time was observed using this recombinant FVII. CONCLUSION: As far as we are aware, this is the first report of expression of recombinant FVII fused with a his-tag through Gateway technology. The next steps, including large scale expression, purification, activation and stabilisation, are underway. PMID- 20011644 TI - Legal guardians and refusal of blood transfusion. PMID- 20011646 TI - Lenalidomide-induced acute liver failure. PMID- 20011645 TI - Recommendations for the use of antithrombin concentrates and prothrombin complex concentrates. PMID- 20011649 TI - Perspective on the use of LNT for radiation protection and risk assessment by the U.S. Environmental Protection Agency. AB - The U.S. Environmental Protection Agency (EPA) bases its risk assessments, regulatory limits, and nonregulatory guidelines for population exposures to low level ionizing radiation on the linear no-threshold (LNT) hypothesis, which assumes that the risk of cancer due to a low dose exposure is proportional to dose, with no threshold. The use of LNT for radiation protection purposes has been repeatedly endorsed by authoritative scientific advisory bodies, including the National Academy of Sciences' BEIR Committees, whose recommendations form a primary basis of EPA's risk assessment methodology. Although recent radiobiological findings indicate novel damage and repair processes at low doses, LNT is supported by data from both epidemiology and radiobiology. Given the current state of the science, the consensus positions of key scientific and governmental bodies, as well as the conservatism and calculational convenience of the LNT assumption, it is unlikely that EPA will modify this approach in the near future. PMID- 20011650 TI - Investigation of non-linear adaptive responses and split dose recovery induced by ionizing radiation in three human epithelial derived cell lines. AB - Two almost completely exclusive fields in radiobiology deal with splitting doses of radiation and comparing the effect to a similar total dose given in one exposure. In radiotherapy, dose "fractionation" is used to "spare" normal tissue and in the low dose field, the adaptive response is well documented as a phenomenon where a small "priming" dose administered before the larger "challenge" dose reduces the effect of the large dose. There have been very few studies where these fields overlap, thus it is not possible to ascertain whether common or distinct mechanisms underlie both phenomena but this is certainly an interesting question and relevant to our understanding of high and low dose radiobiology. This paper presents data for three human cell lines with varying p53 status and radiation responses, treated at a range of times between first and second dose and for 3 different first doses (0.1, 0.5 and 2Gy). The data show that time between doses is critical. Protective (adaptive) effects were seen in each cell line but most prominently in the malignant HT 29 cell line. Surprisingly none of the cell lines showed pronounced split dose recovery. This suggests different mechanisms may underlie the two phenomena. PMID- 20011651 TI - The hormetic morphogen theory of curvature and the morphogenesis and pathology of tubular and other curved structures. AB - In vitro, morphogens such as transforming growth factor (TGF)-beta can up-and down-regulate cell growth at low and high concentrations respectively, i.e. they behave like hormetic agents. The hormetic morphogen theory of curvature proposes that in vivo tissue gradients of such morphogens secreted by source cells determine the fate of cells within their gradient fields (field cells) and that morphogen-induced amplitude modulation of field cell mitochondrial adenosine triphosphate (ATP) generation controls field cell growth along the morphogen gradients: At the high concentration end of gradients, field cell ATP generation and field cell growth is reduced. With declining concentrations along the rest of the gradients field cell ATP and growth is progressively less reduced until an equidyne point is reached, beyond which ATP generation and growth gradually increases. Thus, the differential growth rates along the gradients curve the tissue. Apoptosis at very high morphogen concentrations enables lumen and cavity formation of tubular, spherical, cystic, domed, and other curved biological structures. The morphogen concentration, the gradient slope and the hormesis responses of field cells determine the curvature of such structures during developmental morphogenesis, tissue remodeling and repair of injury. Aberrant hormetic morphogen signaling is associated with developmental abnormalities, vascular diseases, and tumor formation. PMID- 20011652 TI - Calculating hematopoietic-mode-lethality risk avoidance associated with radionuclide decorporation countermeasures related to a radiological terrorism incident. AB - This paper provides theoretical health-risk-assessment tools that are designed to facilitate planning for and managing radiological terrorism incidents that involve ingestion exposure to bone-seeking radionuclides (e.g., radiostrontium nuclides). The focus is on evaluating lethality risk avoidance (RAV; i.e., the decrease in risk) that is associated with radionuclide decorporation countermeasures employed to remove ingested bone-seeking beta and/or gamma emitting radionuclides from the body. To illustrate the application of tools presented, hypothetical radiostrontium decorporation scenarios were considered that involved evaluating the hematopoietic-mode-lethality RAV. For evaluating the efficacy of specific decorporation countermeasures, the lethality risk avoidance proportion (RAP; which is the RAV divided by the total lethality risk in the absence of protective countermeasures) is introduced. The lethality RAP is expected to be a useful tool for designing optimal radionuclide decorporation schemes and for identifying green, yellow and red dose-rate zones. For the green zone, essentially all of the lethality risk is expected to be avoided (RAP = 1) as a consequence of the radionuclide decorporation scheme used. For the yellow zone, some but not all of the lethality risk is expected to be avoided. For the red zone, none of the lethality risk (which equals 1) is expected to be avoided. PMID- 20011654 TI - Lenalidomide, melphalan, and prednisone association is an effective salvage therapy in relapsed plasma cell leukaemia. AB - Plasma cell leukemia (PCL) is a rare and aggressive plasma cell disorder, characterized by the presence of a peripheral blood absolute plasma cell count of at least 2 x 109/l and more than 20% circulating plasma cells. The prognosis of PCL patients remains poor. Even by using autologous or allogenic transplant procedures, median survival does not exceed 3 years (Saccaro et al., 2005). Thalidomide, bortezomib and lenalidomide (Revlimid) have emerged as high active agents in the treatment of PCL (Johnston and abdalla, 2002; Musto et al., 2007; Finnegan et al., 2006). In particular, Lenalidomide is a structural analogue of thalidomide with similar but more potent biological activity; it is used as first line therapy in MM (Palumbo et al., 2007; Niesvizky et al., 2007), although information regarding its associated use with dexamethasone use as salvage therapy in PCL derives from anecdotal single case reports (Musto et al., 2008). We would like to describe a case of primary PCL with adverse cytogenetic in which excellent response was achieved with the combination of lenalidomide, melphalan, and prednisone as salvage therapy. PMID- 20011653 TI - Biphasic dose response in low level light therapy. AB - The use of low levels of visible or near infrared light for reducing pain, inflammation and edema, promoting healing of wounds, deeper tissues and nerves, and preventing cell death and tissue damage has been known for over forty years since the invention of lasers. Despite many reports of positive findings from experiments conducted in vitro, in animal models and in randomized controlled clinical trials, LLLT remains controversial in mainstream medicine. The biochemical mechanisms underlying the positive effects are incompletely understood, and the complexity of rationally choosing amongst a large number of illumination parameters such as wavelength, fluence, power density, pulse structure and treatment timing has led to the publication of a number of negative studies as well as many positive ones. A biphasic dose response has been frequently observed where low levels of light have a much better effect on stimulating and repairing tissues than higher levels of light. The so-called Arndt-Schulz curve is frequently used to describe this biphasic dose response. This review will cover the molecular and cellular mechanisms in LLLT, and describe some of our recent results in vitro and in vivo that provide scientific explanations for this biphasic dose response. PMID- 20011655 TI - Cytokine expression in CD3+ cells in an infant with food protein-induced enterocolitis syndrome (FPIES): case report. AB - Food protein-induced enterocolitis syndrome (FPIES) is a non-IgE-mediated food allergy characterized by severe vomiting, diarrhea, and often failure to thrive in infants. Symptoms typically resolve after the triggering food-derived protein is removed from the diet and recur within few hours after the re-exposure to the causal protein. The diagnosis is based on clinical symptoms and a positive food challenge. In this study, we report a case of FPIES to rice in an 8-month-old boy. We performed a double-blind placebo-controlled food challenge (DBPCFC) to rice and we measured the intracellular T cell expression of interleukin-4 (IL-4); IL-10, and interferon gamma (IFN-gamma) pre-and post-challenge during an acute FPIES reaction and when tolerance to rice had been achieved. For the first time we describe an increase in T cell IL-4 and decrease in IFN-gamma expression after a positive challenge with rice (i.e. rice triggered a FPIES attack) and an increase in T cell IL-10 expression after rice challenge 6 months later after a negative challenge (i.e., the child had acquired tolerance to rice) in an 8 month old with documented FPIES to rice. A Th2 activation associated with high IL-4 levels may contribute to the pathophysiology of the disease. On the other hand, T cell-derived IL-10 may play a role in the acquisition of immunotolerance by regulating the Th1 and Th2 responses. PMID- 20011656 TI - A general total variation minimization theorem for compressed sensing based interior tomography. AB - Recently, in the compressed sensing framework we found that a two-dimensional interior region-of-interest (ROI) can be exactly reconstructed via the total variation minimization if the ROI is piecewise constant (Yu and Wang, 2009). Here we present a general theorem charactering a minimization property for a piecewise constant function defined on a domain in any dimension. Our major mathematical tool to prove this result is functional analysis without involving the Dirac delta function, which was heuristically used by Yu and Wang (2009). PMID- 20011657 TI - Fasting Upregulates PPARalpha Target Genes in Brain and Influences Pituitary Hormone Expression in a PPARalpha Dependent Manner. AB - PPARalpha is a lipid-activable transcription factor that mediates the adaptive response to fasting. Recent data indicate an important role of brain PPARalpha in physiological functions. However, it has not yet been shown whether PPARalpha in brain can be activated in the fasting state. Here we demonstrate that fasting of rats increased mRNA concentrations of typical PPARalpha target genes implicated in beta-oxidation of fatty acids (acyl-CoA oxidase, carnitine palmitoyltransferase-1, medium chain acyl-CoA dehydrogenase) and ketogenesis (mitochondrial 3-hydroxy-3-methylglutaryl-CoA synthase) in pituitary gland and partially also in frontal cortex and diencephalon compared to nonfasted animals. These data strongly indicate that fasting activates PPARalpha in brain and pituitary gland. Furthermore, pituitary prolactin and luteinizing hormone-beta mRNA concentrations were increased upon fasting in wild-type mice but not in mice lacking PPARalpha. For proopiomelanocortin and thyrotropin-beta, genotype specific differences in pituitary mRNA concentrations were observed. Thus, PPARalpha seems to be involved in transcriptional regulation of pituitary hormones. PMID- 20011658 TI - Biomarkers: a definite plus in pneumonia. AB - During the past few years, biomarkers have emerged as an indispensible tool in the diagnosis of pneumonia. To find an ideal diagnostic biomarker for pneumonia is not an easy task. Not only should it allow an early diagnosis of the condition, but it should also allow differential diagnosis from other noninfectious conditions. Ongoing research is being done in this field so as to put an array of biomarkers at the disposal of doctors to improve the diagnosis of pneumonia when patients present to them with cough or nonspecific symptoms which could easily be misinterpreted as symptoms of other conditions. Procalcitonin and soluble triggering receptor expressed on myeloid cells-1 have emerged as reliable diagnostic markers in pneumonia, and are better when compared to other markers, namely, C-reactive protein, leukocyte count, and proinflammatory cytokines. Many other biomarkers are being studied for their probable use in diagnosing pneumonia but have yet to prove their benefit. PMID- 20011659 TI - Toward the discovery of vaccine adjuvants: coupling in silico screening and in vitro analysis of antagonist binding to human and mouse CCR4 receptors. AB - BACKGROUND: Adjuvants enhance or modify an immune response that is made to an antigen. An antagonist of the chemokine CCR4 receptor can display adjuvant-like properties by diminishing the ability of CD4+CD25+ regulatory T cells (Tregs) to down-regulate immune responses. METHODOLOGY: Here, we have used protein modelling to create a plausible chemokine receptor model with the aim of using virtual screening to identify potential small molecule chemokine antagonists. A combination of homology modelling and molecular docking was used to create a model of the CCR4 receptor in order to investigate potential lead compounds that display antagonistic properties. Three-dimensional structure-based virtual screening of the CCR4 receptor identified 116 small molecules that were calculated to have a high affinity for the receptor; these were tested experimentally for CCR4 antagonism. Fifteen of these small molecules were shown to inhibit specifically CCR4-mediated cell migration, including that of CCR4(+) Tregs. SIGNIFICANCE: Our CCR4 antagonists act as adjuvants augmenting human T cell proliferation in an in vitro immune response model and compound SP50 increases T cell and antibody responses in vivo when combined with vaccine antigens of Mycobacterium tuberculosis and Plasmodium yoelii in mice. PMID- 20011660 TI - ActRIIA and BMPRII Type II BMP receptor subunits selectively required for Smad4 independent BMP7-evoked chemotaxis. AB - Bone morphogenetic protein (BMP)-evoked reorientation and chemotaxis of cells occurs with rapid onset and involves events local to the cell membrane. The signaling pathways underlying these rapid processes likely diverge from those mediating classical transcriptional responses to BMPs but it remains unclear how BMP receptors are utilized to generate distinct intracellular mechanisms. We show that BMP7-evoked chemotaxis of monocytic cells depends on the activity of canonical type II BMP receptors. Although the three canonical type II BMP receptors are expressed in monocytic cells, inhibition of receptor subunit expression by RNAi reveals that ActRIIA and BMPRII, but not ActRIIB, are each essential for BMP7-evoked chemotaxis but not required individually for BMP mediated induction. Furthermore, the chemotactic response to BMP7 does not involve canonical Smad4-dependent signaling but acts through PI3K-dependent signaling, illustrating selective activation of distinct intracellular events through differential engagement of receptors. We suggest a model of a BMP receptor complex in which the coordinated activity of ActRIIA and BMPRII receptor subunits selectively mediates the chemotactic response to BMP7. PMID- 20011661 TI - Nuclear phosphatidylcholine and sphingomyelin metabolism of thyroid cells changes during stratospheric balloon flight. AB - Nuclear sphingomyelin and phosphatidylcholine metabolism is involved in the response to ultraviolet radiation treatment in different ways related to the physiological state of cells. To evaluate the effects of low levels of radiation from the stratosphere on thyroid cells, proliferating and quiescent FRTL-5 cells were flown in a stratospheric balloon (BIRBA mission). After recovery, the activity of neutral sphingomyelinase, phosphatidylcholine-specific phospholipase C, sphingomyelin synthase, and reverse sphingomyelin synthase was assayed in purified nuclei and the nuclei-free fraction. In proliferating FRTL-5, space radiation stimulate nuclear neutral sphingomyelinase and reverse sphingomyelin synthase activity, whereas phosphatidylcholine-specific phospholipase C and sphingomyelin synthase were inhibited, thus inducing sphingomyelin degradation and phosphatidylcholine synthesis. This effect was lower in quiescent cells. The possible role of nuclear lipid metabolism in the thyroid damage induced by space radiations is discussed. PMID- 20011662 TI - DNA, RNA, and protein extraction: the past and the present. AB - Extraction of DNA, RNA, and protein is the basic method used in molecular biology. These biomolecules can be isolated from any biological material for subsequent downstream processes, analytical, or preparative purposes. In the past, the process of extraction and purification of nucleic acids used to be complicated, time-consuming, labor-intensive, and limited in terms of overall throughput. Currently, there are many specialized methods that can be used to extract pure biomolecules, such as solution-based and column-based protocols. Manual method has certainly come a long way over time with various commercial offerings which included complete kits containing most of the components needed to isolate nucleic acid, but most of them require repeated centrifugation steps, followed by removal of supernatants depending on the type of specimen and additional mechanical treatment. Automated systems designed for medium-to-large laboratories have grown in demand over recent years. It is an alternative to labor-intensive manual methods. The technology should allow a high throughput of samples; the yield, purity, reproducibility, and scalability of the biomolecules as well as the speed, accuracy, and reliability of the assay should be maximal, while minimizing the risk of cross-contamination. PMID- 20011663 TI - Production of polyclonal antibody against interleukin-33 and assessment of its distribution in murine liver and lung. AB - Interleukin (IL)-33 is the latest member of IL-1 cytokine family. In this study, the cloning, expression, purification, and polyclonal antibody preparation of mouse IL-33 were described. The coding region of IL-33 mature protein was cloned into the prokaryotic expression vector pET-44. The recombinant protein, IL-33 containing a hexahistidine tag in the C-terminal, was expressed in Escherichia coli. The expressed soluble protein was purified by immobilized metal-ion affinity chromatography using Ni2+-nitrilotriacetic acid agarose. The rabbits were immunized with the purified recombinant protein. The obtained antiserum was precipitated by saturated ammonium sulfate and then purified by Protein A affinity chromatography. The sensitivity and specificity of the antibodies were evaluated by enzyme-linked immunosorbent assay and immunohistochemistry. The high titer (1 : 32000) polyclonal antibodies with high specificity were obtained by immunizing rabbits with the purified recombinant protein. Significant expression of IL-33 was seen in mouse liver and lung tissues determined with the anti-IL-33. The production of the polyclonal antibody against IL-33 provides a good tool for studying the biofunctions of IL-33. PMID- 20011664 TI - Postirradiation sarcoma: clinicopathologic features and role of chemotherapy in the treatment strategy. AB - Purpose. An analysis of the clinicopathologic features and treatment of patients was performed to guide evaluation and management of postirradiation sarcoma. Patients and Methods. Between 1994 and 2001, 25 patients with postirradiation sarcoma were treated in one center with different chemotherapy, mainly in neoadjuvant setting (19). Tumors for which these patients received radiotherapy initially were mainly breast carcinoma (for 15 patients). The postirradiation sarcomas were of different histopathologic forms, most frequently osteosarcoma, leiomyosarcoma, and angiosarcoma. Results. Of the 25 patients, 19 were initially treated with chemotherapy. Nine of 19 pretreated patients achieved clinical partial response (RP = 47%). Leiomyosarcomas were good responders (3/4) and undifferentiated sarcoma (3/5). Responders were more often treated with MAID (6/8). Eight of the 9 responders underwent surgery. Two patients achieved complete histological response. Seven of the 9 good responders are alive with a median follow up of 24 months. For all treated patients, median follow up 24 months (6-84 months), overall survival and disease free survival were, respectively, 17/25 (68%), and 14/25 (56%). Conclusion. From our data, postirradiation sarcoma should not be managed differently from primary sarcoma. Chemotherapy has to be included in the treatment plan of postirradiation sarcoma, in future studies. PMID- 20011665 TI - WHEN PARENTS WITH SEVERE MENTAL ILLNESS LOSE CONTACT WITH THEIR CHILDREN: ARE PSYCHIATRIC SYMPTOMS OR SUBSTANCE USE TO BLAME? AB - This study compared parental psychiatric symptom severity, and the absence or presence of severe substance abuse, as predictors of contact with minor children for a representative sample of adults with diagnoses of serious mental illness (N = 45). Child contact and psychiatric symptom severity were measured during regularly scheduled 6-month research interviews over a total 30-month period following each participant's entry into the project. Severe substance abuse was documented as present or absent for the 6-month interval preceding each interview. Results revealed that incidence of severe substance abuse was repeatedly associated with less frequent parent-child contact, even after controlling for psychiatric symptoms, diagnosis, gender, age, ethnicity, and socioeconomic status. Neither psychiatric diagnosis nor symptom severity predicted frequency of child contact when substance abuse was taken into account. Mental health agencies offering parenting classes for adults with serious mental illness should incorporate substance use interventions to reduce loss of child custody and strengthen parent-child relationships. PMID- 20011666 TI - The Minimization of Public Health Risks in Newspapers after Hurricane Katrina. AB - During natural disasters, mass media facilitate the timely provision of accurate information about health risks to the public. This study informs our understanding of such public health discourse utilizing content-analysis of 235 newspaper articles in four major metropolitan newspapers published in the five weeks after Hurricane Katrina hit the Gulf coast in August 2005. These data reveal a small and diminishing number of articles included public health information over time, detailed the hurricane impact on affected communities, and used reliable health sources. The implications for future research from a public health and media relations perspective are discussed. PMID- 20011667 TI - Theoretical Considerations for Understanding "Understanding" by Adults With Right Hemisphere Brain Damage. AB - This article reviews and evaluates leading accounts of narrative comprehension deficits in adults with focal damage to the right cerebral hemisphere (RHD). It begins with a discussion of models of comprehension, which explain how comprehension proceeds through increasingly complex levels of representation. These models include two phases of comprehension processes, broad activation of information as well as pruning and focusing interpretation of meaning based on context. The potential effects of RHD on each processing phase are reviewed, focusing on factors that range from relatively specific (e.g., how the right versus the left hemisphere activate word meanings; how the right hemisphere is involved in inferencing) to more general (the influence of cognitive resource factors; the role of suppression of contextually-irrelevant information). Next, two specific accounts of RHD comprehension difficulties, coarse coding and suppression deficit, are described. These have been construed as opposing processes, but a possible reconciliation is proposed related to the different phases of comprehension and the extent of meaning activation. Finally, the article addresses the influences of contextual constraint on language processing and the continuity of literal and nonliteral language processing, two areas in which future developments may assist our clinical planning. PMID- 20011668 TI - Facile and General Method for Synthesis of Sugar Coated Gold Nanoparticles. AB - This letter describes a general method for the preparation of carbohydrate coated gold nanoparticles. The generality of this method has been demonstrated by surface coating AuNPs with the following sugars: glucose (monosaccharide); sucrose, maltose, or lactose (disaccharides); raffinose (trisaccharide); and starch (polysaccharide). The non-toxic, water-soluble phosphino aminoacid P(CH(2)NHCH(CH(3)-)COOH)(3), THPAL, has been used as a reducing agent in this process. The sizes of sugar coated AuNPs that have been generated in this study are: 30 +/- 8 nm (Glucose), 10 +/- 6 nm (sucrose), 8 +/- 2 nm (maltose), 3 +/- 1 nm (lactose), 6 +/- 2 nm (raffinose), and 39 +/- 9 nm (starch). PMID- 20011669 TI - Parents' Campaigns to Reduce their Children's Conduct Problems: Interactions with Temperamental Resistance to Control. AB - Longitudinal studies have found associations between parenting and the development of conduct problems, and have found that resistant to control temperament moderates these associations. Intervention studies have found associations between intervention-induced changes in parenting and subsequent reductions in children's conduct problems. However, no study to date has evaluated whether parents' self-initiated efforts to change their parenting practices affect children's conduct problems and whether effects depend on children's temperament. The current study asked whether parents' concerted efforts, or campaigns, to increase their involvement and limit-setting were effective in reducing growth in conduct problems from late childhood to early adolescence. It also asked whether the effects of campaigns varied according to children's levels of temperamental resistance to control. Analyses statistically controlled for parenting practices and conduct problems before the campaigns, socioeconomic status, gender, and ethnicity. Results indicated that campaigns that included increased involvement and limit-setting were beneficial only for youths who were rated in early childhood as temperamentally resistant to control. PMID- 20011670 TI - A MOLECULAR EXAMINATION OF RELATEDNESS, MULTIPLE PATERNITY, AND COHABITATION OF THE SOUTHERN PLAINS WOODRAT (NEOTOMA MICROPUS). AB - Two hundred twenty-two individuals of the southern plains woodrat (Neotoma micropus) were captured from 198 excavated middens at 10 discrete collecting sites from a single population in south-central Texas. Field data, mitochondrial D-loop haplotypes, and polymorphic microsatellite loci (5-7) were used to determine genetic patterns in parentage, relatedness, and mating strategy. Microsatellite loci were highly polymorphic (average observed heterozygosity = 0.859) and were used to construct genotypes that were unique for each individual (probability of identical genotypes: 1 in 2,104,567). Results indicated a high frequency of multiple paternity (6 of 9 litters), evidence of repeat mating between the same 2 individuals, and no indication of male dominance at any collection site. Examination of these data suggested a promiscuous mating system. Within a site, average relatedness between adult females was similar to that between adult males. A higher level of cohabitation from that previously documented was recorded and finer-scale analyses revealed high levels of relatedness between most cohabiting individuals. Taken with results from other studies of mating behaviors of N. micropus, our results suggest that mating and social behavior of this species are likely influenced by population density. PMID- 20011671 TI - Dead tired and bone weary: Grandmothers as caregivers in drug affected inner city households. AB - At a time of unprecedented growth in the numbers of custodial grandparents, this case study of Emma's household articulates the stresses inherent to the lives of many grandparents whose own children's lives are governed by drug use and addiction. We contrast normative expectations traditionally integral to the culture of extended families with the counternormative demands that drug use imposes on households. This highlights the untenable nature of caregiving for Emma and countless others of her generation. Compelled by tradition and sentiment to help their own children, they are thus allowing drug use driven norms, values and beliefs to permeate the lives of the grandchildren in their care. Yet, they are also trying to protect those children from drugs and from the violence and conflict that drugs bring into the household. Emma's own life illustrates the salience of norms of kinship, reciprocity and respect, and the trauma in her household demonstrates how their absence does, indeed, intensify demands and erode resources. We conclude that the imperatives of raising the next generation may necessitate a counternormative willingness on the part of grandparents to exclude their adult drug using children from their households. PMID- 20011672 TI - The structure of teenage employment: Social background and the jobs held by high school seniors. AB - Although it is widely assumed that work careers begin after the completion of schooling, most enrolled high school students are also workers. Teenage workers are heavily concentrated in the low wage service sector, but they are also found as supplemental part-time workers in many occupations, including clerical, retail sales, and blue collar employment. Gender, race/ethnicity, and socioeconomic origins are important determinants of the types of jobs that teenage students hold. Students from advantaged socioeconomic origins and students with above average grades are more likely to work in "good jobs," defined by lower hours of work per week and higher status. PMID- 20011673 TI - Implementation of a Serial Replica Exchange Method in a Physics-Based United Residue (UNRES) Force Field. AB - The kinetic-trapping problem in simulating protein folding can be overcome by using a Replica Exchange Method (REM). However, in implementing REM in molecular dynamics simulations, synchronization between processors on parallel computers is required, and communication between processors limits its ability to sample conformational space in a complex system efficiently. To minimize communication between processors during the simulation, a Serial Replica Exchange Method (SREM) has been proposed recently by Hagan et al. (J. Phys. Chem. B2007, 111, 1416 1423). Here, we report the implementation of this new SREM algorithm with our physics-based united-residue (UNRES) force field. The method has been tested on the protein 1E0L with a temperature-independent UNRES force field and on terminally blocked deca-alanine (Ala(10)) and 1GAB with the recently introduced temperature-dependent UNRES force field. With the temperature-independent force field, SREM reproduces the results of REM but is more efficient in terms of wall clock time and scales better on distributed-memory machines. However, exact application of SREM to the temperature-dependent UNRES algorithm requires the determination of a four-dimensional distribution of UNRES energy components instead of a one-dimensional energy distribution for each temperature, which is prohibitively expensive. Hence, we assumed that the temperature dependence of the force field can be ignored for neighboring temperatures. This version of SREM worked for Ala(10) which is a simple system but failed to reproduce the thermodynamic results as well as regular REM on the more complex 1GAB protein. Hence, SREM can be applied to the temperature-independent but not to the temperature-dependent UNRES force field. PMID- 20011674 TI - Assessment of preschoolers' positive empathy: concurrent and longitudinal relations with positive emotion, social competence, and sympathy. AB - The purpose of this study was to examine a new measure of children's dispositional positive empathy (i.e., reactions to others' positive emotions) and its concurrent and longitudinal relations with positive emotion, social competence, and empathy/sympathy with negative emotions. At Time 1, 192 3.5-year olds (88 girls) participated; at Time 2, 1 year later, 168 4.5-year-olds (79 girls) participated. Children's positive empathy was reported by mothers and observed in the laboratory at Time 2. Additionally, mothers, fathers, and non parental caregivers completed questionnaires at Time 1 and Time 2 regarding children's positive emotion, empathy/sympathy, and social competence. Children's positive emotion was observed at both assessments. There was evidence of reliability of the new reported measure of positive empathy. Additionally, there were numerous positive relations between positive empathy and social competence and between positive empathy and empathy/sympathy with negative emotions. This study provides unique insight into children's positive empathy and relations to socio-emotional functioning. PMID- 20011675 TI - Ortho-quinone-enhanced ascorbate oxidation. Combined roles of lipid charge and the magnesium cation. AB - Quinones are widely distributed compounds in nature. Of these, ortho-quinones are found to be involved in the pathogenic mechanism of Parkinson's disease, in oxidative deaminations to free-radical redox reactions, and as intermediates in the pathways implicated in the carcinogenicity of 2,3- and 3,4-catechol estrogens. Addition of MgCl(2) to solutions of the hydrophobic ortho-quinones, 1,10-phenanthroquinone (PHQ) and beta-lapachone (LQ) enhances ascorbate oxidation in the absence or presence of large unilamellar vesicles (LUVs) of the neutral lipid dimyristoylphos-phatidylcholine (DMPC), although initial rates of ascorbate oxidation are smaller in the presence of lipid as compared to its absence. Addition of this salt to solutions of the para-quinone 1,4-naphthoquinone (NQ) did not affect the ascorbate rate of oxidation in the absence or presence of DMPC. Addition of MgCl(2) to semiquinone solutions of PHQ or LQ in the presence or absence of DMPC increases semiquinone stability, as detected from the semiquinone disproportionation equilibrium displacement to semiquinone formation. Furthermore, MgCl(2) increases the partition of the ortho-semiquinones into the aqueous phase, although no such effect is observed for the semiquinone of NQ. For all the quinones under study, smaller rates of ascorbate oxidation and of semiquinone equilibrium concentration occur in the presence of negatively charged LUVs composed of an equimolar mixture of DMPC and dimyristoylphosphatidic acid DMPA. Ascorbate oxidation rate enhancements correlate with an increase in semiquinone concentration with addition of MgCl(2), in the absence or presence of neutral lipid. This observation favors the proposition that ascorbate oxidation rate increases are caused by semiquinone thermodynamic stabilization. Thus, the ascorbate oxidation rate enhancement by MgCl(2) in solutions containing hydrophobic ortho-quinones is still possible in systems with hydrophobic environments analogous to that of DMPC. PMID- 20011676 TI - Modeling Search for People in 900 Scenes: A combined source model of eye guidance. AB - How predictable are human eye movements during search in real world scenes? We recorded 14 observers' eye movements as they performed a search task (person detection) in 912 outdoor scenes. Observers were highly consistent in the regions fixated during search, even when the target was absent from the scene. These eye movements were used to evaluate computational models of search guidance from three sources: saliency, target features, and scene context. Each of these models independently outperformed a cross-image control in predicting human fixations. Models that combined sources of guidance ultimately predicted 94% of human agreement, with the scene context component providing the most explanatory power. None of the models, however, could reach the precision and fidelity of an attentional map defined by human fixations. This work puts forth a benchmark for computational models of search in real world scenes. Further improvements in modeling should capture mechanisms underlying the selectivity of observer's fixations during search. PMID- 20011677 TI - Subjective versus Objective Definitions of Homelessness: Are there Differences in Risk Factors among Heavy-Drinking Women? AB - Investigations with homeless populations have focused on those living on the streets or in shelters; few have examined phenomena based on respondents' self definitions as homeless or not. This investigation examined similarities and differences among risk factors (including mental health, substance abuse, religion/spirituality, social support, and risky sexual behaviors) using two definitions of homelessness: one where place of residence defined individuals as homeless (the 'objective', or traditional, definition) and another where respondents defined themselves as homeless (the 'subjective' definition). Data come from the baseline survey of the NIAAA-funded "Sister-to-Sister" study (n=339) of heavy-drinking women. Subjectively-defined homelessness was associated with higher rates of mental health and substance use disorders, lower rates of condom use, higher rates of trading sex for food, and less social support. Objectively-defined homelessness was associated with higher rates of drinking in abandoned buildings, on the streets, and in public restrooms, more new sexual partners, and higher rates of trading sex for heroin and speedballs. Investigations failing to ask for subjective information may misattribute some factors to homelessness which may overestimate the effect of various factors on homelessness. Investigators should ask respondents to define their homelessness, or they lose an important dimension of the concept of homelessness. PMID- 20011678 TI - Who Receives Confrontation in Recovery Houses and when is it Experienced as Supportive? AB - The Alcohol and Drug Confrontation Scale (ADCS) is a 72-item instrument that measures a construct of confrontation defined as warnings about potential harm associated with alcohol and drug use. This analysis describes the characteristics of individuals entering residential recovery homes (N=323) who received confrontation and when it was experienced as supportive. A large proportion reported receiving at least one confrontational statement (80%), most commonly from family/friends (71%). Individuals who did and did not receive confrontation did not differ by demographics, but those receiving confrontation had more recent substance use, higher perceived costs of sobriety and more severe family and psychiatric problems. Differences were noted in confrontation from the criminal justice system versus family/friends. Overall, residents experienced confrontation as supportive regardless of who confronted them. Residents who experienced confrontation the most helpful were those with higher levels of substance use and those who believed maintaining sobriety would be difficult. PMID- 20011679 TI - An Analysis of Teacher Investment in the Context of a Family-School Intervention for Children with ADHD. AB - Intervention researchers have often failed to assess treatment integrity; when integrity is examined, the focus is typically on whether the steps of intervention have been applied and not on quality of implementation. In the few studies that have investigated intervention quality, the emphasis has been on how intervention is delivered and not how it is received or the degree of participant engagement. This study was designed to examine participant engagement, specifically teacher investment, in the context of family interventions for children with attention-deficit/hyperactivity disorder (ADHD) that were linked with the school. The interventions included Family-School Success (FSS), a program that engages families and schools in a problem solving partnership, and Coping with ADHD through Relationships and Education (CARE), a program designed to provide education and support to families. Participants included the families and teachers of children in grades 2 through 6; 45 sets of families and teachers participated in FSS and 48 were in CARE. The Teacher Investment Questionnaire (TIQ) was designed to assess teacher engagement in intervention. The TIQ is a rating scale for clinicians to be completed during and after treatment. The findings provided support for the reliability and validity of the TIQ. Level of teacher involvement was demonstrated to decline with advancing grade level for FSS, but not CARE. Parent ratings of the quality of the family-school relationship as well as the level of teacher support for homework, assessed at baseline, were shown to be significantly related to clinician ratings of teacher investment post intervention. The findings highlight the importance of assessing participant engagement in intervention. Strategies to refine the assessment of teacher investment were discussed. PMID- 20011681 TI - Promoting Academic Success Among Latino Youth. AB - This article describes results from the Oregon Latino Youth Survey, which was designed to identify factors that promoted or hindered academic success for Latino middle- and high-school youngsters. The study samples included a total of 564 Latino and non-Latino students and parents. Analyses showed that Latino students reported a high frequency of discriminatory experiences and institutional barriers at school, and that they and their parents were more likely to experience institutional barriers compared to non-Latinos. Further, Latino students and parents reported that they/their youngsters were more likely to dropout of school compared to non-Latinos. Path models showed lower acculturation and more institutional barriers were related to less academic success for Latino students. More parent academic encouragement and staff extracurricular encouragement were associated with better academic outcomes for Latino students. Finally, family socioeconomic disadvantage had an indirect effect on Latino youngster academic success, through effects on parent monitoring and school involvement. PMID- 20011680 TI - Are There Separate Neural Systems for Spelling? New Insights into the Role of Rules and Memory in Spelling from Functional Magnetic Resonance Imaging. AB - How do people spell the thousands of words at the tips of their tongues? Are words with regular sound-to-letter correspondences (e.g., "blink") spelled using the same neural systems as those with irregular correspondences (e.g., "yacht")? By offering novel neuroimaging evidence, we aim to advance contemporary debate about whether people use a single lexical memory process or whether dual mechanisms of lexical memory and sublexical phonological rules work in concert. We further aim to advance understanding of how people read by taking a fresh look at the related yet distinct capacity to spell. During functional magnetic resonance imaging scanning, 12 participants heard low-frequency regular words, irregular words, and nonwords (e.g., "shelm") and responded whether a visual presentation of the word was spelled correctly or incorrectly. While behavioral measures suggested some differences in accuracy and reaction time for the different word types, the neuroimaging results alone demonstrated robust differential processing and support a dual-route model of spelling, with implications for how spelling is taught and remediated in clinical and educational contexts. PMID- 20011682 TI - Utilizing Social Work Skills to Enhance Entrepreneurship Training for Women: A Ghanaian Perspective. AB - This article describes a women's entrepreneurship exchange program that was designed and facilitated with the cooperation of various governmental and nongovernmental entities in Ghana for Ghanaian women. The article briefly reviews the entrepreneurship development literature from an international perspective and discusses the Entrepreneurship Program as a targeted approach for empowering and sustaining women's economic situation in Ghana. Emphasis is placed on understanding the impact of cultural and social networks and the women's ability to succeed as entrepreneurs through the use of social work skills. PMID- 20011683 TI - Activity-based mass spectrometric characterization of proteases and inhibitors in human saliva. AB - Proteases present in oral fluid effectively modulate the structure and function of some salivary proteins and have been implicated in tissue destruction in oral disease. To identify the proteases operating in the oral environment, proteins in pooled whole saliva supernatant were separated by anion-exchange chromatography and individual fractions were analyzed for proteolytic activity by zymography using salivary histatins as the enzyme substrates. Protein bands displaying proteolytic activity were particularly prominent in the 50-75 kDa region. Individual bands were excised, in-gel trypsinized and subjected to LC/ESI-MS/MS. The data obtained were searched against human, oral microbial and protease databases. A total of 13 proteases were identified all of which were of mammalian origin. Proteases detected in multiple fractions with cleavage specificities toward arginine and lysine residues, were lactotransferrin, kallikrein-1, and human airway trypsin-like protease. Unexpectedly, ten protease inhibitors were co identified suggesting they were associated with the proteases in the same fractions. The inhibitors found most frequently were alpha-2-macroglobulin-like protein 1, alpha-1-antitrypsin, and leukocyte elastase inhibitor. Regulation of oral fluid proteolysis is highly important given that an inbalance in such activities has been correlated to a variety of pathological conditions including oral cancer. PMID- 20011684 TI - Differential response of antioxidant enzymes to cadmium stress in tolerant and sensitive cell line of cucumber (Cucumis sativus L.). AB - Previously, a stable cell suspension of cucumber tolerant to 100 microM CdCl(2) was obtained (Gzyl & Gwozdz, 2005, Plant Cell Tissue Organ Cult 80: 59-67). In this study, the relationship between the activity of antioxidant enzymes and cadmium tolerance of cucumber cells was analyzed. A cadmium-sensitive and the cadmium-tolerant cell lines were exposed to 100 microM and 200 microM CdCl(2) and the activities of superoxide dismutase (SOD), catalase (CAT), ascorbate peroxidase (APOX) and guaiacol peroxidase (POX) were determined. In the sensitive cell line, a decrease of total activity of SOD and POX was observed, whereas the activity of CAT and APOX significantly increased in metal-supplemented medium. By contrast, in the tolerant cells, the total activity of antioxidant enzymes decreased (SOD, CAT) or was maintained at approximately the same level (APOX, POX). Moreover, a different pattern of isoenzyme activity was observed in the tolerant and sensitive cells. These results suggest that an enhanced activity of antioxidant enzymes is not directly involved in the increased tolerance to cadmium of the selected cucumber cell line. PMID- 20011685 TI - Staphylococcus aureus as an infectious agent: overview of biochemistry and molecular genetics of its pathogenicity. AB - Although it is estimated that 20-30% of the general human population are carriers of Staphylococcus aureus, this bacterium is one of the most important etiological agents responsible for healthcare-associated infections. The appearance of methicillin resistant S. aureus (MRSA) strains has created serious therapeutical problems. Detailed understanding of the mechanisms of S. aureus infections seems necessary to develop new effective therapies against this pathogen. In this article, we present an overview of the biochemical and genetic mechanisms of pathogenicity of S. aureus strains. Virulence factors, organization of the genome and regulation of expression of genes involved in virulence, and mechanisms leading to methicilin resistance are presented and briefly discussed. PMID- 20011686 TI - Cardioprotective effect of 5-lipoxygenase gene (ALOX5) silencing in ischemia reperfusion. AB - It is well known that 5-lipoxygenase derivates of arachidonic acid play an important pathogenic role during myocardial infarction. Therefore, the gene encoding arachidonate 5-lipoxygenase (ALOX5) appears to be an attractive target for RNA interference (RNAi) application. In experiments on cultivated cardiomyocytes with anoxia-reoxygenation (AR) and in vivo using rat model of heart ischemia-reperfusion (IR) we determined influence of ALOX5 silencing on myocardial cell death. ALOX5 silencing was quantified using real-time PCR, semi quantitative PCR, and evaluation of LTC(4) concentration in cardiac tissue. A 4.7 fold decrease of ALOX5 expression (P < 0.05) was observed in isolated cardiomyocytes together with a reduced number of necrotic cardiomyocytes (P < 0.05), increased number live (P < 0.05) and unchanged number of apoptotic cells during AR of cardiomyocytes. Downregulation of ALOX5 expression in myocardial tissue by 19% (P < 0.05) resulted in a 3.8-fold reduction of infarct size in an open chest rat model of heart IR (P < 0.05). Thus, RNAi targeting of ALOX5 protects heart cells against IR injury both in culture and in vivo. PMID- 20011688 TI - An integrated approach to the understanding of Chlamydia infections: Introduction to the 2009 update. AB - An integrated, multidisciplinary effort is needed to achieve a greater understanding of the immunopathogenesis of urogenital and ocular Chlamydia trachomatis infections to combat this infection successfully, to prevent long term complications and to develop a vaccine. The contribution of molecular epidemiology and host-pathogen genomics to the understanding of diseases caused by C. trachomatis was explored in depth by the ICTI Consortium, which was first introduced in the Netherlands in December 2004 at the First Annual Amsterdam Chlamydia Meeting (AACM) and was the subject of a special Drugs of Today supplement published in 2006, entitled "An integrated approach to the understanding of Chlamydia infections". The ICTI Consortium formed the basis for a larger, European Union-funded international effort involving 20 centers, the EpiGenChlamydia Consortium. Major themes, including epidemiology, genetics and immunogenetics, biology and immunology, involving the chlamydiae species infecting humans, such as C. trachomatis, C. psittaci and C. pneumoniae, predominantly presented at the 4th and 5th AACM, are summarized in this second Drugs of Today supplement, as well as timely mini-reviews of the advances made in the field. PMID- 20011689 TI - The EU FP6 EpiGenChlamydia Consortium: contribution of molecular epidemiology and host-pathogen genomics to understanding Chlamydia trachomatis-related disease. AB - Chlamydia trachomatis infections are responsible for the world's leading cause of blindness (trachoma) and its most prevalent sexually transmitted disease, which is strongly associated with pelvic inflammatory disease, ectopic pregnancy and tubal infertility. Twin study-based findings of members of EpiGenChlamydia Consortium estimate that there is a 40% genetic predisposition to C. trachomatis infections. It is likely that the advances in human genomics will help to unravel the genetic predisposition at the gene level and will help to define a genetic fingerprint that can be used as a marker for this predisposition. The information gathered to date suggests that this predisposition and the factors contributing to prognosis are multifactorial. The EpiGenChlamydia Consortium aims to structure transnational research to such a degree that comparative genomics and genetic epidemiology can be performed in large numbers of unrelated individuals. Biobanking and data-warehouse building are the most central deliverables of the Coordination Action of the Consortium in Functional Genomics Research. In addition, the collective synergy acquired in this Coordination Action will allow for the generation of scientific knowledge on the C. trachomatis-host interaction, knowledge on the genetic predisposition to C. trachomatis infection and the development of tools for early detection of a predisposition to C. trachomatis infection and its complications. This review summarizes the consortium aims and progress, and future perspectives and directions. PMID- 20011690 TI - An integrated approach to Chlamydia trachomatis infection: the ICTI Consortium, an update. AB - The desire for an integrated approach to the study of Chlamydia trachomatis infection of the female genital tract resulted in the creation of the "ICTI Consortium" (Integrated approach to the study of Chlamydia Trachomatis Infection) in 2005. The ICTI consortium is based on tight interaction and collaboration between basic scientists, clinicians, epidemiologists and health-care policy makers. This translational approach will help to further the valuable insight into the immunopathogenesis of this sexually transmitted infection in order to develop new intervention strategies, including the vaccines and screening programs necessary to effectively diagnose, treat and prevent C. trachomatis infection. This article provides an update of ICTI Consortium activities carried out until September 2009. PMID- 20011691 TI - Chlamydia trachomatis today: treatment, detection, immunogenetics and the need for a greater global understanding of chlamydial disease pathogenesis. AB - Chlamydia trachomatis is an important human pathogen causing a myriad of severe and debilitating diseases. While antibiotics have been a mainstay of treatment, there is increasing evidence for potential drug resistance, reinfection and persistent infections that require a reevaluation of treatment strategies. A critical need to address these issues will be a rapid, sensitive and cost effective diagnostic that can be used for global screening, treatment and test-of cure of infected individuals instead of empirical therapy that not only drives drug resistance but is not costeffective. This type of diagnostic would allow clinicians and researchers to evaluate the true incidence and prevalence of chlamydial infections in both developed and developing countries. There are extremely limited data on chlamydial sexually transmitted diseases (STDs) in many developing countries, including those in Central and South America. In addition, advancing our understanding of chlamydial disease pathogenesis will require an evaluation of host genetic susceptibility to infection and sequelae. We provide preliminary data on rates of chlamydial STDs and host genetic factors that predispose to infection among adolescent pregnant and nonpregnant commercial sex worker populations residing in Quito, Ecuador. PMID- 20011692 TI - Chlamydia trachomatis serovar distributions in Russian men and women: a comparison with Dutch serovar distributions. AB - The data on serovar distributions of Chlamydia trachomatis - the most diagnosed sexually transmitted infection (STI) worldwide - are important for epidemiologic purposes and transmission studies but are completely lacking in Russia. The aim of the current study is to determine the serogroup and serovar distributions in Russian men and women and compare these data with Dutch serogroup and serovar distributions. In Russian men and women, serogroup B was the most prevalent (46%), followed by the intermediate serogroup (I group; 33%) and serogroup C (21%). The distribution was comparable between men and women. The serogroup distribution was similar to the previously published distribution in Dutch cohorts. However, on a serovar level statistically very significant differences were observed, reaching up to P < 0.0001. The serovars B and G/Ga had higher prevalences compared with the reported Dutch prevalences, while serovars F, H, I/Ia, J and K had lower prevalences compared with the Dutch studies. In conclusion, this is the first report of Russian C. trachomatis serovar/serogroup distributions. Serogroup B is the most prevalent, followed by serogroup I and serogroup C with no statistical differences on the serogroup level. However, significant differences between Russia and the Netherlands were observed in the distribution of C. trachomatis serovars. PMID- 20011693 TI - Lymphogranuloma venereum, where do we stand?: clinical recommendations. AB - Lymphogranuloma venereum (LGV) was formerly known as a sexually transmissible infection confined to equatorial regions, but also as an "imported" sexually transmissible infection in the Western world. However, since 2003, with the first cases of LGV proctitis among men who have sex with men reported in the Netherlands, an ongoing epidemic has been revealed in Western society dating back to at least 1981. In this article a case of LGV proctitis is presented, and the state of the art diagnostics, treatment and common complications concerning LGV are discussed. Moreover, risk factors and the background of the recent epidemic of LGV in the Western world among men who have sex with men are summarized. There is a need to develop new diagnostic assays, to prevent complications and to protect the community from more expansive transmission. Shorter antibiotic treatment courses for LGV are necessary but require large controlled clinical trials. The microbial and immunological background of LGV infection in relation to HIV should be studied in detail and could help to explain the considerable number of asymptomatic LGV cases. PMID- 20011694 TI - Host genetic contribution to the cellular immune response to Chlamydia trachomatis: Heritability estimate from a Gambian twin study. AB - If the cellular immune response to Chlamydia trachomatis is subject to genetic influences, the degree and mechanisms of such genetic control may have important implications for vaccine development. We estimated the relative contribution of host genetics to the total variation in lymphoproliferative responses to C. trachomatis antigen by analyzing these responses in 64 Gambian twin pairs from trachoma endemic areas. Zygosity was determined by restriction fragment length polymorphism analysis of minisatellite probes and microsatellite typing. Proliferative responses to serovar A elementary body antigen were estimated in monozygotic (MZ) and dizygotic (DZ) twin pairs. We found a stronger correlation and lower within-pair variability in these responses in MZ than in DZ twin pairs. The heritability estimate was 0.39 (P = 0.07) suggesting that host genetic factors contributed 39% of the variation. A better understanding of these genetic influences will contribute to the elucidation of preventive therapies for ocular C. trachomatis infection and may identify important mechanisms in protection for rational vaccine construction. PMID- 20011695 TI - Identification of candidate genes using the murine model of female genital tract infection with Chlamydia trachomatis. AB - The integrated approach to the study of female genital tract infection (GTI) with Chlamydia trachomatis is a conceptual framework through which a consistent and comprehensive evidence-based understanding of C. trachomatis GTI could evolve. One application of this approach has been to identify candidate genes that may play a role in the course and severity of C. trachomatis GTI in women, using human clinical and genetic data together with results obtained in the female mouse model to guide the selection process This model has been proven robust enough: i) to identify stable phenotypic differences in the course and outcome of GTI among commonly used immunocompetent inbred mouse stains that are used in the construction of gene knockout (KO) and transgenic mice; as well as ii) to serve as a platform in which to assess the influence of genetic differences among human genital tract isolates of C. trachomatis as well as between this biovar and the mouse biovar, Chlamydia muridarum. This review presents a summary of published and unpublished results from 25 years of studies in immunodeficient and gene deficient KO mice that both inform our present understanding of the immunogenetics of C. trachomatis GTI and serve to guide candidate gene selection. PMID- 20011696 TI - Host genetic studies in human ocular Chlamydial infection. AB - Several human and animal models and methods have been used to dissect genetic contributions to immunity and pathogenesis of chlamydial diseases. Considerable achievements have been made in this field of host genetics. The hope is that these studies will lead to medical applications by helping to elicit the function of genes that are involved in host defense against chlamydia and in progression to severe sequelae. In the present article, we review a selection of findings in the forward genetics of ocular Chlamydia trachomatis infection in humans. PMID- 20011697 TI - TLR2 haplotypes in the susceptibility to and severity of Chlamydia trachomatis infections in Dutch women. AB - Chlamydia trachomatis infections may cause several disease conditions ranging from asymptomatic infections to severe upper genital tract pathology, thereby causing significant morbidity worldwide. Remarkable interindividual differences in the clinical course of C. trachomatis infection have been observed, and are mainly based on variation in genes encoding immune-regulatory and bacteria sensing proteins. Toll-like receptors (TLRs) are closely involved in pathogen recognition and host defense in C. trachomatis infections. The aim of this study is to assess the role of TLR2 single nucleotide polymorphisms and haplotypes in the susceptibility to, and severity of C. trachomatis infections. The study comprised a sexually transmitted disease cohort of 468 Dutch Caucasian women and a control group of 321 women. The subfertility cohort consisted of 56 women with clinically well-defined tubal pathology. The results showed no significant differences in individual TLR2 genotype frequencies in the susceptibility for C. trachomatis infections between the C. trachomatis-positive group and controls. However, haplotype 1 was statistically significant (P = 0.015) and was associated with protection against tubal pathology following C. trachomatis infection. The same haplotype was also significantly decreased (P = 0.021) in increasing severity of C. trachomatis infections (asymptomatic > symptomatic > tubal pathology) suggesting a protective effect of this haplotype against the development of late complications. PMID- 20011698 TI - TLR4 in Chlamydia trachomatis infections: knockout mice, STD patients and women with tubal factor subfertility. AB - Chlamydia trachomatis is the most prevalent sexually transmitted bacterium in the world with almost 100 million new cases each year, some of which will develop tubal pathology. Clear differences in its clinical course of infections have been observed, and recently it has been shown that 40% is based on host genetic factors. We used an integrated approach based on infection of Toll-like receptor 4 (TLR4) knockout mice and immunogenetic analysis of female sexually transmitted disease (STD) patients (susceptibility) and women with C. trachomatis-associated tubal factor subfertility (severity). The results in TLR4 knockout mice suggest that the protection against reinfection is more solid in normal as compared to the TLR4-deficient mice. In humans the functional TLR4 single nucleotide polymorphism studied was not involved in the susceptibility to infection. However, C. trachomatis immunoglobulin (Ig) G-positive subfertile women with tubal pathology were more than twice as likely to be carriers of the mutant TLR4 +896 G allele as compared to those without tubal pathology; however this observation did not reach statistical significance. In conclusion, both the murine model and the human immunogenetics studies show a slight effect upon TLR4 deficiency in the severity of infection but not in the susceptibility to infection. PMID- 20011699 TI - TLR9 KO mice, haplotypes and CPG indices in Chlamydia trachomatis infection. AB - Previous studies have investigated the role of Toll-like receptor (TLR)2 and TLR4 in susceptibility to and severity of Chlamydia trachomatis infections. In this study we employ a unique integrated approach to study the role of the intracellular CpG DNA receptor: we use a murine knockout (KO) model to assess TLR9 relevance, study human TLR9 genotypes and haplotypes in sexually transmitted disease (STD) patients and subfertile women with or without tubal pathology and use in silico TLR9 CpG index calculations to assess potential immunostimulatory properties of the Chlamydia bacterium. Although no significant differences in the course of initial infections were observed between KO mice and wild-type mice the TLR9 KO mice showed a significant level of protection upon reinfection (P = 0.02). We did not observe significant differences in genotype frequencies between C. trachomatis-positive and C. trachomatisnegative women (STD patients). However, haplotype analyses revealed a trend between C. trachomatis-positive and C. trachomatis-negative women in the carriage of haplotype IV (P = 0.061; OR: 2.6; 95% CI: 1.0-6.8). In women with subfertility, odds ratios between 2 and 3 were found for tubal pathology risk, but they did not reach significance due to cohort size limitations. Finally, CpG sequence analysis showed mildly immunostimulatory properties for the genomic sequences of Chlamydia serovars B and D. Based on the murine model, human immunogenetic studies and in silico CpG index analyses, TLR9 seems to play a modest role in C. trachomatis infections. Extension of the human cohorts is necessary to significantly prove the effect in humans. PMID- 20011700 TI - Analyses of polymorphisms in the inflammasome-associated NLRP3 and miRNA-146A genes in the susceptibility to and tubal pathology of Chlamydia trachomatis infection. AB - Susceptibility to Chlamydia trachomatis infections is 40% host based. microRNA 146a is a negative regulator of Tolllike receptor (TLR) signaling and possesses functional polymorphisms which decrease the production of premiR-146a and mature miR-146a. Single nucleotide polymorphisms (SNPs) in NLRP3 are associated with decreased NLRP3 expression and hypoproduction of interleukin (IL)-1beta. We investigated whether the SNPs miR-146a G>C (rs2910164), NLRP3 C>T (rs4925663) and G>A (rs12065526) are associated with the susceptibility to and severity of C. trachomatis infection. The genotypes of three SNPs were tested in two cohorts: cohort 1 consists of Dutch women (n = 318) attending a sexually transmitted disease (STD) clinic and cohort 2 (n = 277) consists of subfertile (n = 184) and healthy Finnish women (n=93). While in cohort 1 the analyzed SNPs were not associated with the susceptibility to C. trachomatis infections (C. trachomatis positive vs. C. trachomatis-negative), we showed in C. trachomatis-positive women that the NLRP3 mutant AG and AA genotypes were a risk factor for the development of symptoms (P = 0.047, OR = 2.9) and more specifically for having lower abdominal pain (genotype AA: P = 0.022, OR = 31.3). In the Finnish tubal pathology group versus the control group no statistical significant differences in the incidences of the SNPs studied were found, nor for the degree of tubal pathology. In conclusion, the mutant NLRP3 A allele is a risk factor for the development of symptoms, specifically lower abdominal pain, after a C. trachomatis infection in women attending an STD clinic. PMID- 20011702 TI - A new avenue to investigate: the autophagic process. From Crohn's disease to Chlamydia. AB - The finding that a variant (T300A) of the autophagyrelated 16-like 1 (ATG16L1) gene is associated with Crohn's disease suggests that the inability to eliminate intestinal intracellular microbes via (macro)autophagy may be involved in the pathogenesis of this disease. The variant induces an autophagy-associated defect in Paneth cells, specialized cells in the crypts of Lieberkuhn within the small intestine that secrete defensins and other antimicrobial peptides. Moreover, other loci, IRGM and LRRK2 involved in autophagy and implicated in clearance of intracellular bacteria have been found to be associated with Crohn's disease. These unexpected findings have changed the focus of research in Crohn's disease and have stimulated an in-depth study of the complex process of autophagy. Autophagy is regulated by many genes and is emerging as a central player in the immunologic control of intracellular bacteria. Chlamydia trachomatis is able to inhibit apoptosis and the production of nuclear factor kappa B (NFkappaB) in order to survive in the host. Extensive studies on association of genes regulating the inflammatory response in experimental models and in humans as revised in other sections of this supplement have failed to explain the longterm complications of C. trachomatis infection. The advances in the molecular pathways of Chlamydia infection and their effects on the Golgi apparatus and other cytoplasmic organelles suggests that defects in autophagic genes may predispose the host to chronic infection and be responsible for the long-term complications. A new genomic approach of the complete autophagic pathway may reveal new insight to understand the presence of a complication in affected individuals, even if at present there is no evidence that C. trachomatis is affected by this pathway. PMID- 20011701 TI - Inflammasomes bridge signaling between pathogen identification and the immune response. AB - Microbial organisms express pathogen-associated molecular patterns (PAMPs) that can stimulate expression of proinflammatory mediators following ligation of pathogen recognition receptors. However, both commensal organisms and pathogens can express PAMPs. The immune system can distinguish between commensals and pathogens in part through secretion of the key inflammatory cytokines interleukin (IL)-1beta and IL-18. A PAMP such as lipopolysaccharide can induce production of intracellular pro-IL-1beta and pro-IL-18, but not their secretion. A second "danger signal", derived from host-cell molecules that are released from stressed or infected cells, or detected as a PAMP that is present in the cytosol, can stimulate assembly of an inflammasome that activates the protease caspase-1. Caspase-1, in turn, is responsible for processing and secretion of the mature IL 1beta and IL-18. Many diverse ligands leading to inflammasome activation have been identified, but the cell signaling pathways initiated by the ligands tend to converge on a small set of common mechanisms. PMID- 20011703 TI - Aspects of reproductive biology that influence the distribution and spread of Chlamydia trachomatis within the female genital tract: a new paradigm. AB - Critical to evaluating Chlamydia trachomatis vaccine candidates is the availability of appropriate animal models. At a minimum, models must mimic the essential features of transmission and disease progression that contribute to the severe outcomes associated with upper genital tract infection. Existing models, whether mouse, pig or nonhuman primate, are based on the generally accepted premise that upper genital tract infection, when it occurs, is an event subsequent to cervical infection. However, what this simple paradigm overlooks are many features of reproductive biology that could influence both the initial distribution and subsequent spread of C. trachomatis within the female genital tract, as well as the immune responses made at these site(s) of infection. A review of the literature strongly suggests that the menstrual cycle and coitusrelated phenomena are likely to have a profound effect on the course and outcome of female genital tract infection with C. trachomatis. Although the new paradigm that emerges raises concerns about the adequacy of existing animal models, it also suggests ways to modify these models to better mimic the complexities of human infection and therefore serve as appropriate models in which to test the safety and efficacy of vaccine candidates against C. trachomatis infection in women. PMID- 20011704 TI - CXCL13 expression in Chlamydia trachomatis infection of the female reproductive tract. AB - Chlamydia trachomatis is the most common cause of acute salpingitis worldwide. The socioeconomic impact of sexually transmitted infections (STI) caused by C. trachomatis is considerable. The purpose of this study was to investigate secretion of a unique chemokine, CXCL13, during the inflammatory process in human fallopian tube tissue in response to infection with C. trachomatis. We employed two models for our experiments: archived fallopian tube paraffin sections from known cases of salpingitis of unknown etiology and human fallopian tube organ culture established from fresh fallopian tube biopsies subsequently infected in vitro with C. trachomatis serovar E. We used immunohistochemistry, microarray analysis and cytometric bead array to study these specimens. In both models, we found that the fallopian tissue infected with C. trachomatis expressed CXCL13 and other characteristics of tertiary lymphoid tissue. In addition, we found that CXCL13 was expressed in multiple cell types, including endothelial cells, demonstrating a mechanism for the lymphoid aggregation seen in fallopian tube tissue during salpingitis and infection with C. trachomatis. PMID- 20011705 TI - Significantly higher serologic responses of Chlamydia trachomatis B group serovars versus C and I serogroups. AB - Chlamydia trachomatis serovars are divided into three serogroups, namely serogroup B, serogroup I (Intermediate) and serogroup C, and subsequently into 19 different serovars. Worldwide, serogroup B is the most prevalent followed by serogroup I. Clear differences have been observed in the duration of infection and growth kinetics between serovars from different serogroups in murine and cell culture models. Reasons for these observed differences are bacterial and host related, and are not well understood. The aim of this study was to determine the differences in immunoglobulin (Ig) G responses between the three serogroups in a group of patients infected with different serovars. Serovars were assessed from 235 C. trachomatispositive patients and quantitative IgG responses were determined. Analyses of variance were used to compare the IgG responses between the three serogroups. Of the serovars, 46% were B group (with serovar E the most prevalent: 35.3%), 39.6% were I group and 14.3% were C group. A highly significant difference in serologic response was shown when comparing the mean IgG concentrations (AU/mL) of patients having serovars in the most prevalent serogroup compared to the other serogroups: B = 135, C = 46 and I = 60 (B vs. C and B vs. I, P < 0.001). In conclusion, the most prevalent serovars generate the highest serologic responses. PMID- 20011706 TI - From disease to etiology: historical aspects of Chlamydia-related diseases in animals and humans. AB - A first description of the etiology of trachoma was published in 1907. During expeditions to Java to study the transmission of syphilis, Halberstaedter and von Prowazek infected orangutans with conjunctival scrapings from trachoma patients and such agents in conjunctival smears. They called them "chlamydozoa". J. Ritter in 1879 published the first case description of psittacosis, describing a mini epidemic in which three of seven patients died, and identified the source of infection, determined the incubation period and the nontransmissibility of the disease from human to human. In 1895 the term psittacosis was first applied. In 1893, Nocard isolated a Gram-negative bacterium from parrots dying of psittacosis (Bacillus psittacosis). This organism was subsequently found in human or avian subjects and was later diagnosed as Salmonella. The inconsistent bacteriological findings prompted a search for a filterable virus during the pandemic of 1929 1930. Almost simultaneously, Levinthal (1930), Coles (1930) and Lillie (1930) described small, filterable bodies in infectious material called "Levinthal-Coles Lillie (L.C.L.) bodies". Bedson first suggested the biphasic development cycle in 1932 after having studied tissues from inoculated mice. In 1935, Burnet and Rountree propagated "the virus" on the chorioallantoic membrane of embryonated chicken. PMID- 20011707 TI - Chlamydophila psittaci infections in turkeys: overview of economic and zoonotic importance and vaccine development. AB - We provide evidence on the multifactorial infectious etiology of respiratory disease in turkeys. Although Chlamydophila psittaci is difficult to diagnose, this entity should not be neglected in veterinary diagnostic laboratories. The present results suggest a pathogenic interplay between chlamydophila, avian metapneumovirus and Ornithobacterium rhinotracheale. Additionally, we demonstrate zoonotic transmission from turkeys to humans. Psittacosis due to contact with poultry probably occurs more often than is thought and the infection can be asymptomatic or symptomatic. There is no commercial C. psittaci vaccine available and currently the best option is an experimental major outer membrane protein based DNA vaccine. PMID- 20011708 TI - Chlamydophila psittaci infections in The Netherlands. AB - Psittacosis, caused by Chlamydophila psittaci, is a well described but sporadically occurring clinical entity, which mainly presents as community acquired pneumonia. Diagnosis used to be relatively difficult. However, new molecular techniques, such as real-time polymerase chain reaction, increased detection of cases. Furthermore, genotyping of the ompA gene can be used as a tool to trace the possible source of an outbreak or to link a specific bird to a particular patient. PMID- 20011709 TI - The inflammatory aspects of Chlamydia pneumoniae-induced brain infection. AB - The importance of inflammation in the pathogenesis of neurodegenerative diseases, such as Alzheimer's disease, is increasingly being recognized. Although amyloid beta is considered to be one of the main initiators of these inflammatory processes, some reports suggest that brain infections may also contribute or even initiate the neuroinflammation. One of the best studied pathogens that might be involved in this phenomenon is the herpes simplex virus, but more recently the obligate intracellular respiratory Gram-negative bacterium, Chlamydia pneumoniae, has also been associated with Alzheimer's disease. The present article discusses recent data on the role of C. pneumoniae infection in neuroinflammation and its potential contribution to the pathogenesis of Alzheimer's disease. PMID- 20011710 TI - Exploring the role of Chlamydia pneumoniae in cardiovascular disease: a narrative review. AB - An overwhelming number of studies have suggested that Chlamydia pneumoniae infections play a role in the development of atherosclerosis. Several, but not all, seroepidemiological studies have shown that C. pneumoniae antibodies may be related to the development of atherosclerotic disease. Additionally, C. pneumoniae seems to be present in atherosclerotic but not in healthy vascular tissue. Experimental studies have suggested a number of molecular mechanisms by which vascular C. pneumoniae infection might stimulate atherosclerosis development. Alternatively, nonvascular C. pneumoniae infection may cause clinically relevant atherosclerosis-related cardiovascular events through the systemic effects of chronic infection. Genetic variation may account for individual differences in susceptibility to the proatherogenic effects of C. pneumoniae infection. Despite the suggested role of infection in atherosclerosis, antibiotics seem to have no place in the secondary prevention of atherosclerosis related cardiovascular events. The present narrative review evaluates the role of C. pneumoniae infections in the development of cardiovascular disease. PMID- 20011711 TI - Bioinformatic approaches to the study of Chlamydial diseases. AB - The combination of bioinformatics, genomics and proteomics is a rapidly expanding research field that has now become essential for the investigation of biological, cellular and molecular phenomena, both in terms of established facts and in prediction models based on simulations, and therefore can greatly contribute to the development of potential new therapeutic approaches. This article provides numerous, well-known examples illustrating the importance of bioinformatics in modern biological and medical research in the field of infectious diseases, specifically Chlamydia trachomatis infection. Specialized chlamydia databases and analysis programs and the information they contain provide the opportunity for understanding and identifying key processes and phenomena associated with the unique infection cycle of chlamydia, and are paving the way towards the development of novel diagnostic and therapeutic tools. Bioinformatics may provide the clues necessary for a true breakthrough in the fight against chlamydial diseases: the development of a vaccine. PMID- 20011712 TI - Dynamic kine magnetic resonance imaging in whiplash patients and in age- and sex matched controls. AB - The multitude of symptoms following a whiplash injury has given rise to much discussion because of the lack of objective radiological findings. The ligaments that stabilize the upper cervical spine can be injured. Dynamic kine magnetic resonance imaging (dMRI) may reveal the pathological motion patterns caused by injury to these ligaments. To compare the findings and motion patterns in the upper cervical spine, 25 whiplash trauma patients with longstanding pain, limb symptoms and loss of balance indicating a problem at the level of C0-C2, as well as matched healthy controls were imaged using dMRI. Imaging was performed with an Intera 1.5 T (Philips Healthcare, USA) magnet. A physiotherapist performed the bending and rotation of the upper cervical spine for the subjects to ensure that the movements were limited to the C0-C2 level. An oblique coronal T2- and proton density-weighted sequence and a balanced fast field echo axial sequence were used. The movements between C0-C2 and the signal from the alar ligaments were analyzed. Contact of the transverse ligament and the medulla in rotation was seen in two patients. The signal from the alar ligaments was abnormal in 92% of the patients and in 24% of the control subjects (P<0.0001). Abnormal movements at the level of C1-C2 were more common in patients than in controls (56% versus 20%, P=0.028). Whiplash patients with longstanding symptoms had both more abnormal signals from the alar ligaments and more abnormal movements on dMRI at the C0-C2 level than controls. PMID- 20011713 TI - Reliability of temporal summation and diffuse noxious inhibitory control. AB - BACKGROUND: The test-retest reliability of temporal summation (TS) and diffuse noxious inhibitory control (DNIC) has not been reported to date. Establishing such reliability would support the possibility of future experimental studies examining factors affecting TS and DNIC. Similarly, the use of manual algometry to induce TS, or an occlusion cuff to induce DNIC of TS to mechanical stimuli, has not been reported to date. Such devices may offer a simpler method than current techniques for inducing TS and DNIC, affording assessment at more anatomical locations and in more varied research settings. METHOD: The present study assessed the test-retest reliability of TS and DNIC using the above techniques. Sex differences on these measures were also investigated. RESULTS: Repeated measures ANOVA indicated successful induction of TS and DNIC, with no significant differences across test-retest occasions. Sex effects were not significant for any measure or interaction. Intraclass correlations indicated high test-retest reliability for all measures; however, there was large interindividual variation between test and retest measurements. CONCLUSION: The present results indicate acceptable within-session test-retest reliability of TS and DNIC. The results support the possibility of future experimental studies examining factors affecting TS and DNIC. PMID- 20011714 TI - A survey of prelicensure pain curricula in health science faculties in Canadian universities. AB - OBJECTIVE: The present exploratory, descriptive study aimed to determine the designated time for mandatory pain content in curricula of major Canadian universities for students in health science and veterinary programs before being licensed. METHOD: Major Canadian university sites (n=10) were chosen where health science faculties included at least medicine (n=10) and nursing (n=10); many also included dentistry (n=8), pharmacy (n=7), physical therapy (n=8) and/or occupational therapy (n=6). These disciplines provide the largest number of students entering the workforce but are not the only ones contributing to the health professional team. Veterinary programs (n=4) were also surveyed as a comparison. The Pain Education Survey, developed from previous research and piloted, was used to determine total mandatory pain hours. RESULTS: The majority of health science programs (67.5%) were unable to specify designated hours for pain. Only 32.5% respondents could identify specific hours allotted for pain course content and/or additional clinical conferences. The average total time per discipline across all years varied from 13 h to 41 h (range 0 h to 109 h). All veterinary respondents identified mandatory designated pain content time (mean 87 h, range 27 h to 200 h). The proportion allotted to the eight content categories varied, but time was least for pain misbeliefs, assessment and monitoring/follow up planning. CONCLUSIONS: Only one-third of the present sample could identify time designated for teaching mandatory pain content. Two-thirds reported 'integrated' content that was not quantifiable or able to be determined, which may suggest it is not a priority at that site. Many expressed a need for pain related curriculum resources. PMID- 20011715 TI - Does classification of persons with fibromyalgia into Multidimensional Pain Inventory subgroups detect differences in outcome after a standard chronic pain management program? AB - INTRODUCTION: The present study aimed to replicate and validate the empirically derived subgroup classification based on the Multidimensional Pain Inventory (MPI) in a sample of highly disabled fibromyalgia (FM) patients. Second, it examined how the identified subgroups differed in their response to an intensive, interdisciplinary inpatient pain management program. METHODS: Participants were 118 persons with FM who experienced persistent pain and were disabled. Subgroup classification was conducted by cluster analysis using MPI subscale scores at entry to the program. At program entry and discharge, participants completed the MPI, Medical Outcomes Study Short Form-36, Hospital Anxiety and Depression Scale and Coping Strategies Questionnaire. RESULTS: Cluster analysis identified three subgroups in the highly disabled sample that were similar to those described by other studies using less disabled samples of FM. The dysfunctional subgroup (DYS; 36% of the sample) showed the highest level of depression, the interpersonally distressed subgroup (ID; 24%) showed a modest level of depression and the adaptive copers subgroup (AC; 38%) showed the lowest depression scores in the MPI (negative mood), Medical Outcomes Study Short Form-36 (mental health), Hospital Anxiety and Depression Scale (depression) and Coping Strategies Questionnaire (catastrophizing). Significant differences in treatment outcome were observed among the three subgroups in terms of reduction of pain severity (as assessed using the MPI). The effect sizes were 1.42 for DYS, 1.32 for AC and 0.62 for ID (P=0.004 for pairwise comparison of ID-AC and P=0.018 for ID-DYS). DISCUSSION: These findings underscore the importance of assessing individuals' differences in how they adjust to FM. PMID- 20011716 TI - Factors associated with chronic noncancer pain in the Canadian population. AB - Chronic noncancer pain (CNCP) is a prevalent health problem with pervasive negative effects on the individual's quality of life. Previous epidemiological studies of CNCP have suggested a number of individual biological, psychological and societal correlates of CNCP, but it has rarely been possible to simultaneously compare the relative strengths of many such correlates in a Canadian population sample. With data provided by the 1996/1997 Canadian National Population Health Survey, ordinal logistic regression was used to examine the extent to which a number of population variables are associated with CNCP in a large (n=69,365) dataset. The analysis revealed cross-sectional correlations of varying strengths between CNCP and 27 factors. Increasing age, low income, low educational achievement, daily cigarette smoking, physical inactivity and abstention from alcohol were among the factors found to increase CNCP risk. The considerable impact of distress and depression on CNCP are also highlighted. A number of comorbid medical illnesses increased CNCP risk, including some (such as chronic obstructive pulmonary disease, epilepsy and thyroid disease) that have not hitherto been associated with pain. White race and the affirmation of an important role for spirituality or faith reduced CNCP risk. In contrast to some previous studies, female sex did not emerge as an independent CNCP risk. The present exploratory analysis describes associations between CNCP and a number of characteristics from several domains, thus suggesting many areas for further research. PMID- 20011718 TI - Work-related asthma: A case-based guide. AB - Work-related asthma is common yet underdiagnosed. It is a significant cause of morbidity and socioeconomic loss. Diagnosis is often difficult, and requires a strong index of suspicion and careful investigation. The Canadian Thoracic Society has endorsed the recent American College of Chest Physicians consensus statement on work-related asthma. The present document illustrates the advised approach to diagnosis and management of work-related asthma using case-based examples of occupational asthma and work-exacerbated asthma. The main statements of advice from the American College of Chest Physicians consensus statement are reproduced with permission. PMID- 20011717 TI - Development and psychometric evaluation of a new measure of pain-related support preferences: the Pain Response Preference Questionnaire. AB - BACKGROUND: Behavioural conceptualizations of chronic pain posit that solicitous responses to pain behaviours are positively reinforcing and play a role in the development of chronic pain and disability. Recent research suggests that studies investigating this model were likely limited by the use of only a few narrowly defined categories of responses to pain behaviour. A measure of preferences regarding pain-related social support has the potential to improve behavioural models of chronic pain by identifying other potentially reinforcing responses to pain behaviour. OBJECTIVE: The Pain Response Preference Questionnaire (PRPQ) was created to assess preferences regarding pain-related social support. The purpose of the present study was to empirically develop PRPQ scales and examine their psychometric properties. METHODS: A large university student sample (n=487) free of chronic pain completed the 39-item PRPQ. Factor analysis was applied to the data from the present sample to empirically develop PRPQ scales. Using a second student sample (n=87), relationships between the PRPQ scales and theoretically related measures were examined to evaluate the construct validity of the scales. Factor analysis supported four factors that reflected preferences for emotional and instrumental support, assistance in managing pain and emotions, having one's pain ignored, and being encouraged to persist with one's activities. Based on this analysis, scales labelled solicitude, management, suppression and encouragement were created. Correlation analyses supported the construct validity of these scales. CONCLUSIONS: The PRPQ is a psychometrically sound measure of preferences of pain-related social support. Research with clinical samples is needed to further evaluate its psychometric properties and clinical utility. PMID- 20011719 TI - Canadian Thoracic Society: Presenting a new process for clinical practice guideline production. AB - A key mandate of the Canadian Thoracic Society (CTS) is to promote evidence-based respiratory care through clinical practice guidelines (CPGs). To improve the quality and validity of the production, dissemination and implementation of its CPGs, the CTS has revised its guideline process and has created the Canadian Respiratory Guidelines Committee to oversee this process. The present document outlines the basic methodological tools and principles of the new CTS guideline production process. Important features include standard methods for choosing and formulating optimal questions and for finding, appraising, and summarizing the evidence; use of the Grading of Recommendations Assessment, Development and Evaluation system for rating the quality of evidence and strength of recommendations; use of the Appraisal of Guidelines for Research and Evaluation instrument for quality control during and after guideline development and for appraisal of other guidelines; use of the ADAPTE process for adaptation of existing guidelines to the local context; and use of the GuideLine Implementability Appraisal tool to augment implementability of guidelines. The CTS has also committed to develop guidelines in new areas, an annual guideline review cycle, and a new formal process for dissemination and implementation. Ultimately, it is anticipated that these changes will have a significant impact on the quality of care and clinical outcomes of individuals suffering from respiratory diseases across Canada. PMID- 20011720 TI - Perception of asthma as a factor in career choice among young adults with asthma. AB - BACKGROUND/AIM: Asthma is a common chronic condition that can be aggravated by workplace exposures. Young adults with asthma should know how their future occupation might affect their asthma, and potentially, their quality of life. The aim of the present study was to assess the awareness of young adults to occupational risks for asthma and high-risk occupations, as well as their perception of the role of asthma in career choice. METHODS: Young adults 16 to 22 years of age with reported physician-diagnosed asthma were recruited to complete a questionnaire eliciting information regarding asthma control, career choice and awareness of occupational exposure risks. RESULTS: A small majority of the study cohort (56.4%) could identify occupations that cause or exacerbate asthma, and 34.7% indicated that asthma was an important factor in their career plans. Family physicians were most responsible for asthma management (80.2%), but young adults were more likely to discuss asthma and career plans with their parents (43.6%) or friends (29.7%) than with their family physician (13.9%; P<0.001). CONCLUSION: Young adults with asthma have suboptimal awareness of potential work-related asthma risks. Family physicians most commonly provide asthma care to these young adults. However, few young adults are talking to their family physicians about career choices and asthma. This observation represents an area of asthma care that needs to be explored in young adults with asthma. PMID- 20011721 TI - Pulmonary vein stenosis: Case report and literature review. AB - BACKGROUND: Pulmonary vein stenosis is a rare cause of pulmonary hypertension, with variable onset and presentation. One or more of the four pulmonary veins can be primarily or secondarily affected. A five-month-old girl presented with respiratory distress, lethargy and cyanosis requiring intubation. METHODS: Echocardiography showed right ventricular dilation, a right ventricular systolic pressure of 97 mmHg, decreased ejection fraction and turbulent flow at the left atrium. Cardiac catheterization revealed stenosis of the left-sided pulmonary veins, for which she underwent a Coles procedure. RESULTS: Postoperatively, there was a transient improvement in the patient's pulmonary hypertension but she subsequently deteriorated. Her prognosis was considered bleak, and a decision was made with the family to withdraw care. CONCLUSION: Pulmonary vein stenosis is a rare cause of pulmonary hypertension, and is associated with significant morbidity and mortality. Surgical intervention may be of benefit in selected cases. It stands to reason that any treatment will have the best chance of success if completed before the pulmonary hypertension becomes fixed. Early diagnosis depends on a high index of clinical suspicion. PMID- 20011724 TI - Database epidemiology. PMID- 20011725 TI - Identifying patients with physician-diagnosed asthma in health administrative databases. AB - BACKGROUND: Asthma imposes a heavy and expensive burden on individuals and populations. A population-based surveillance and research program based on health administrative data could measure and study the burden of asthma; however, the validity of a health administrative data diagnosis of asthma must first be confirmed. OBJECTIVE: To evaluate the accuracy of population-based provincial health administrative data in identifying adult patients with asthma for ongoing surveillance and research. METHODS: Patients from randomly selected primary care practices were assigned to four categories according to their previous diagnoses: asthma, chronic obstructive pulmonary disease, related respiratory conditions and nonasthma conditions. In each practice, 10 charts from each category were randomly selected, abstracted, then reviewed by a blinded expert panel who identified them as asthma or nonasthma. These reference standard diagnoses were then linked to the patients' provincial records and compared with health administrative algorithms designed to identify asthma. Analyses were performed using the concepts of diagnostic test evaluation. RESULTS: A total of 518 charts, including 160 from individuals with asthma, were reviewed. The algorithm of two or more ambulatory care visits and/or one or more hospitalization(s) for asthma in two years had a sensitivity of 83.8% (95% CI 77.1% to 89.1%) and a specificity of 76.5% (95% CI 71.8% to 80.8%). CONCLUSION: Definitions of adult asthma using health administrative data are sensitive and specific for identifying adults with asthma. Using these definitions, cohorts of adults with asthma for ongoing population-based surveillance and research can be developed. PMID- 20011726 TI - Comparison of per cent predicted and percentile values for pulmonary function test interpretation. AB - BACKGROUND: Pulmonary function tests (PFTs) are commonly interpreted as a fraction of predicted normal values, with an abnormal test often defined as less than 80% or greater than 120% of the predicted value. However, recommendations of the American Thoracic Society/European Respiratory Society suggest using a percentile-based approach to define an abnormal test (less than the fifth or greater than the 95th percentiles). OBJECTIVE: To compare PFT values obtained by the per cent predicted method with the percentile-based method for lung function parameters. METHODS: Full PFTs performed between January 2000 and July 2004, at the Health Sciences Centre (Winnipeg, Manitoba) were analyzed. Using the Crapo and Gutierrez equations, per cent predicted and percentile values were calculated. An abnormal test was defined as less than 80% or greater than 120% of predicted (per cent predicted method) or as less than the fifth or greater than the 95th percentiles (percentile method). Using the percentile method as reference standard, the diagnostic test characteristics of the per cent predicted method were calculated. RESULTS: The full PFTs of 2176 men and 1658 women were analyzed using the Crapo and Gutierrez equations. The mean (+/- SD) age of all subjects was 52+/-15 years. Per cent agreement between the two tests was more than 94% for all parameters except for reduced residual volume (88%). Per cent predicted methods had suboptimal sensitivity for abnormal total lung capacity (88% to 89%), increased residual volume (83% to 89%) and reduced diffusion capacity (89% with Crapo equations). Suboptimal specificity (83% to 86%) was observed for decreased residual volume. CONCLUSION: The results of the per cent predicted and percentile-based approaches for PFT interpretation were similar for the majority of lung function parameters. These two methods can be used interchangeably for spirometry. However, caution may be warranted in relying solely on per cent predicted methods for assessing lung volume or diffusion capacity. PMID- 20011727 TI - Multiple cavitating nodules in a renal transplant recipient. AB - Pulmonary nodules are common following solid organ transplantation and vary in etiology. Nodules with central cavitation are most likely to be of infectious origin in the post-transplant population. A novel presentation of post-transplant lymphoproliferative disorder manifesting as multiple cavitating pulmonary nodules is described. The patient, a 45-year-old female renal transplant recipient, presented with constitutional symptoms and a chest x-ray showing multiple bilateral cavitating lesions. A computed tomography scan confirmed innumerable, randomly dispersed, cavitating nodules in the lung parenchyma. Multiple large hypodense lesions were identified in the liver and spleen. The appearance of the native and transplanted kidneys was normal. A liver biopsy identified an Epstein- Barr virus-negative, diffuse, large B cell lymphoma. Repeat imaging after treatment with a cyclophosphamide, hydroxydaunorubicin, oncovin and prednisone/prednisolone regimen demonstrated dramatic resolution of all lesions. The present case represents a unique radiographic presentation of post-transplant lymphoproliferative disorder not previously reported in the literature. PMID- 20011729 TI - Are we on track for education and training of future gastroenterologists? Interview by Paul C Adams. PMID- 20011731 TI - Argon photocoagulation in the treatment of gastric antral vascular ectasia and radiation proctitis. AB - Gastric antral vascular ectasia (GAVE) and radiation proctitis are two vascular disorders of the gastrointestinal tract that typically present with recurrent gastrointestinal bleeding. Although the pathogenesis of either condition is not known, they are unlikely to be similar. GAVE appears to be related to autoimmune disorders or cirrhosis, while radiation proctitis is the result of pelvic irradiation, most commonly used for the treatment of pelvic malignancies. Medical therapies for both conditions are not typically effective, and surgical therapies are usually not required because endoscopic treatment, aimed at coagulation of the underlying vascular lesions, has evolved as the most effective therapy. There is limited evidence in the literature for the use of medical and surgical therapies, with most of the evidence coming from case reports involving small numbers of patients. In the present article, we review the evidence for the use of argon plasma photocoagulation (APC, the most commonly used endoscopic modality) in the treatment of GAVE and radiation proctitis. PMID- 20011732 TI - Assessing the educational needs of Canadian gastroenterologists and gastroenterology nurses: challenges to optimal care in Crohn's Disease. AB - OBJECTIVE: A national needs assessment of Canadian gastroenterologists and gastroenterology nurses was undertaken to determine the perceived and unperceived educational and performance barriers to caring for patients with Crohn's disease (CD). METHODS: A triangulated, mixed-method approach (qualitative and quantitative) was used to determine the nature and extent of knowledge gaps and barriers in the care of patients with CD. RESULTS: Qualitative interviews were conducted with nine gastroenterologists, four gastroenterology nurses and nine patients with CD. Based on this exploratory research, a survey was designed and launched nationally (37 gastroenterologists, 36 gastroenterology nurses). Findings indicated that Canadian gastroenterologists and gastroenterology nurses lacked clarity regarding their roles and responsibilities across the continuum of CD care, and face communication gaps within the health care team, undermining their effectiveness. Gastroenterologists identified challenges in optimal diagnosis due to unclear testing and diagnostic criteria. They recognized knowledge gaps when treating patient subgroups and in prescribing biological therapies. Furthermore, gastroenterologists self-identified gaps in skill, knowledge, and confidence in monitoring disease progression and effectively assessing response to therapy. When managing patients with CD, gastroenterologists expressed challenges with patient issues outside their domain of medical expertise, particularly with the skills needed to facilitate effective patient communication and education that would enhance adherence to recommended treatments. CONCLUSIONS: Educational initiatives should address diagnostic and treatment guidelines, as well as enhancement of clinical performance gaps in health care team processes and the patient-professional therapeutic relationship. To impact care and patient outcomes, these initiatives must be relevant to clinical practice settings and applicable to the practice context. PMID- 20011733 TI - Predictors of medication adherence in pediatric inflammatory bowel disease patients at the Stollery Children's Hospital. AB - BACKGROUND: Patients with inflammatory bowel disease (IBD) often do not take their medications as prescribed. OBJECTIVE: To examine self-reported adherence rates in IBD patients at the Stollery Children's Hospital (Edmonton, Alberta) and to determine predictors of medication adherence. METHODS: A survey was mailed to 212 pediatric IBD patients of the Stollery Children's Hospital. A chart review was completed for those who returned the survey. RESULTS: A total of 119 patients completed the survey. The nonresponders were significantly older than responders (14.5 years versus 13.2 years; P=0.032). The overall adherence rate was 80%. Nonadherence was associated with older age (14.6 years versus 13.0 years; P=0.04), longer disease duration (5.0 years versus 3.1 years; P=0.004) and reported use of herbal medications (40.0% versus 13.6%; P=0.029). The most common reasons reported for missing medications were forgetfulness, feeling better and too many medications. In addition, patients reported being more likely to take anti-inflammatory medications and less likely to take herbal medicines. CONCLUSION: Identified predictors of nonadherence such as age, disease duration and use of herbal treatments may enable the development of specific strategies to improve adherence in adolescents with IBD. PMID- 20011734 TI - Prevention of Clostridium difficile infection with Saccharomyces boulardii: a systematic review. AB - BACKGROUND: Clostridium difficile is a major cause of antibiotic-associated diarrhea within the hospital setting. The yeast Saccharomyces boulardii has been found to have some effect in reducing the risk of C difficile infection (CDI); however, its role in preventive therapy has yet to be firmly established. OBJECTIVE: To review the effectiveness of S boulardii in the prevention of primary and recurrent CDI. Benefit was defined as a reduction of diarrhea associated with C difficile. Risk was defined as any adverse effects of S boulardii. METHODS: A literature search in MEDLINE, EMBASE, CINAHL and the Cochrane Library was performed. Included studies were English language, randomized, double-blind placebo controlled trials evaluating S boulardii in CDI prevention. RESULTS: Four studies were reviewed. Two studies investigated the prevention of recurrence in populations that were experiencing CDI at baseline. One trial showed a reduction of relapses in patients experiencing recurrent CDI (RR=0.53; P<0.05). The other demonstrated a trend toward reduction of CDI relapse in the recurrent treatment group of patients receiving high-dose vancomycin (RR=0.33; P=0.05). Two other studies examined primary prevention of CDI in populations that had been recently prescribed antibiotics. These studies lacked the power to detect statistically significant differences. Patients on treatment experienced increased risk for thirst and constipation. CONCLUSION: S boulardii seems to be well tolerated and may be effective for secondary prevention in some specific patient populations with particular concurrent antibiotic treatment. Its role in primary prevention is poorly defined and more research is required before changes in practice are recommended. PMID- 20011736 TI - Fulminant legionellosis in two patients treated with infliximab for Crohn's disease: case series and literature review. AB - Two cases of fulminant pulmonary legionellosis, complicated by prolonged intensive care unit stays and acute respiratory distress syndrome, and who were recently treated with infliximab for Crohn's disease, are described. A review of the literature revealed three additional cases in patients with inflammatory bowel disease, and a total of 22 cases of Legionella pneumophila pneumonia in the context of treatment with antitumour necrosis (TNF)-alpha medications. The median age of the patients was 49 years, and men and women were affected equally. The case fatality rate was 14% (three of 22). Early recognition and treatment of this anti-TNF-alpha-related complication would likely result in reduced mortality and morbidity. Physicians prescribing anti-TNF-alpha drugs should be aware of this association. PMID- 20011735 TI - The role of iron in the pathophysiology and treatment of chronic hepatitis C. AB - Increased hepatic iron content may be observed in patients with chronic hepatitis C infection, and may contribute to disease severity. The presence of hemochromatosis gene mutations is associated with increased hepatic iron accumulation and may lead to accelerated disease progression. Hepatic iron depletion has been postulated to decrease the risk of hepatocellular carcinoma in patients with cirrhosis due to chronic hepatitis C. It is possible that iron depletion stabilizes or improves liver histology and slows disease progression in these individuals. The present article reviews the prevalence and risk factors for hepatic iron overload in chronic hepatitis C, with emphasis on the available data regarding the efficacy of iron depletion in the treatment of this common liver disease. PMID- 20011737 TI - National joint replacement registry. PMID- 20011738 TI - Why do we need a national joint replacement registry in the United States? AB - The continually increasing number of total hip arthroplasties (THAs) being performed, in conjunction with the rapid growth in new surgical techniques and implants related to THA, warrants ongoing and objective monitoring of results. National joint replacement registries have become powerful surveillance systems for monitoring contemporary THAs and improving outcomes. Despite the compelling evidence of their benefits, such a registry has yet to be established in the United States. In this article, we provide a rationale for implementing a national joint replacement registry in the United States. PMID- 20011739 TI - A technique to facilitate everting the patella in stiff or obese knees in total knee arthroplasty. AB - In many techniques for total knee arthroplasty, eversion of the patella is necessary. In stiff or obese knees, eversion is often difficult to perform. With the patella-holding clamp (used to cement the patellar component), it is possible to keep the patella in an everted position while the knee is being flexed. PMID- 20011740 TI - Acetabular component revision in total hip arthroplasty. Part I: cementless shells. AB - Magnitude and location of acetabular bone defects dictate the type of reconstruction required. For the majority of reconstructions, a porous-coated hemispheric shell secured to host bone with multiple screws is the implant of choice. This reconstruction is feasible provided at least 50% of the implant contacts host bone. When such contact is not possible, and there is adequate medial and peripheral bone, techniques using alternative uncemented implants can be used for acetabular reconstruction. An uncemented cup can be placed at a "high hip center." Alternatively, the acetabular cavity can be progressively reamed to accommodate extra-large cups. Oblong cups, which take advantage of the oval shaped cavity resulting from many failed acetabular components, can also be used. The success of these cementless techniques depends on the degree and location of bone loss and on the presence of pelvic discontinuity. PMID- 20011741 TI - Effect of bone cement viscosity and set time on mantle area in total knee arthroplasty. AB - To assess the impact of bone cement viscosity on total knee arthroplasty, we compared 1 high-viscosity and 2 medium-viscosity cements with respect to mantle area and zone-specific intrusion depths into the tibial plateau. We analyzed postoperative radiographs to determine penetration area and depth in 72 consecutive patients (79 knees) in whom DePuy II (n = 11), Endurance (n = 34), or Simplex-P (n = 34) cement was used. Penetration into the tibial plateau (anteroposterior zones 1-4) was significantly reduced with use of the high viscosity DePuy II cement but did not differ significantly between the 2 medium viscosity cements, Endurance and Simplex-P. Surgical and tourniquet times were significantly decreased with the quicker setting DePuy II cement. Given these findings, additional studies are warranted to assess the long-term impact of the lower intrusion depths found with DePuy II cement. Such differences in cement penetration could jeopardize long-term fixation and lead to higher long-term device failure rates. PMID- 20011742 TI - Closed reduction of a dislocated total hip arthroplasty with a constrained acetabular component. PMID- 20011743 TI - Clinical use of porous tantalum in complex primary total knee arthroplasty. PMID- 20011744 TI - Total hip arthroplasty in slipped capital femoral epiphysis: a novel technique to remove embedded knowles pins. AB - The Knowles pin has been well recognized in fixing slipped capital femoral epiphysis. When these pins are left in place for a long period, it is very difficult to remove them during total hip arthroplasty, because the pins have fluted threads, which are oversized in the end and allow bone growth in the previously threaded slots. We present a simple and novel technique to remove Knowles pins so that both unnecessary trauma to the lateral femoral cortex and operating time is reduced significantly. PMID- 20011745 TI - Differences between neurosurgeons and orthopedic surgeons in classifying cervical dislocation injuries and making assessment and treatment decisions: a multicenter reliability study. AB - Variability exists in the management of cervical spinal injuries. The goal of this study was to assess the effect of training specialty (orthopedic surgery vs neurosurgery) on management of cervical dislocations. Twenty-nine spine surgeons reviewed 10 cases of cervical dislocation injuries. For each of the 10 cases, the surgeons evaluated 3 clinical scenarios, which included a neurologically intact patient, a patient with an incomplete spinal cord injury (SCI), and a patient with complete SCI. Surgeons determined whether a unilateral or bilateral facet dislocation was present and whether pretreatment magnetic resonance imaging (MRI) or immediate closed reduction was indicated. Management decisions were re assessed after review of MRIs. While spine surgeons may agree on what they see on MRI and how they classify certain cervical injuries irrespective of training, significant differences of opinion continue to exist regarding the therapeutic implications of this information, specifically, whether to order a pretreatment MRI and how to manage the injury. PMID- 20011746 TI - Traumatic thoracic spondyloptosis without neurologic deficit, and treatment with in situ fusion. PMID- 20011747 TI - Spondylodiscitis after vertebral fracture in the thoracic spine. PMID- 20011748 TI - Professional toothbrushing as a method for diagnosing gingivitis in 3- to 6-year old preschool children. AB - PURPOSE: The aim of the present crossover study was to evaluate professional toothbrushing as a method for diagnosing gingivitis in children. MATERIALS AND METHODS: Thirty-four preschool children who had gingival bleeding > or = 10% and without proximal restorations/carious lesions were included in the study. Examinations comprised two gingival indices recorded at a 15-min interval, in the following sequences: the Ainamo and Bay gingival bleeding index (GBI1) followed by the brushing index (BI2) and vice versa (BI1-GBI2). Half of the children started the study in the first sequence and the other half in the second. After a 3- to 4-day washout period, the indices were again recorded with individuals changing the sequences. Data analysis considered GBI as the gold standard, and sensitivity (SE), specificity (SP), positive predictive value (PPV) and negative predictive value (NPV) were also calculated. RESULTS: The overall mean value of gingival bleeding at the first examination was 18.85 +/- 9.24%. Validation of toothbrushing performed after GBI (BI2) resulted in values for SE, SP, PPV and NPV of 0.59 (95% CI, 0.55 to 0.63), 0.92 (95% CI, 0.91 to 0.93), 0.64 (95% CI, 0.60 to 0.69) and 0.90 (95% CI, 0.89 to 0.92), respectively. BI performed before GBI (BI1) resulted in similar SE, SP, PPV and NPV. Percentage agreement of GBI1 BI2 and BI1-GBI2 was 83.5% and 85.9%, respectively. CONCLUSIONS: Professional toothbrushing can be suggested as a method for the diagnosis of gingival inflammatory status in children, especially as an indicator of gingival health. PMID- 20011749 TI - Differences in oral health among Italian adolescents related to the type of secondary school attended. AB - PURPOSE: The aim of the present study was to evaluate the possible interaction between the type of secondary school attended, oral clinical data and self perceived oral health (SPOH) behaviour among Italian adolescents. MATERIALS AND METHODS: A total of 913 adolescents living in Milan (Italy), attending two different types of secondary schools, grammar and technical secondary schools, were examined regarding dental caries and gingival conditions. Oral health behaviour data were collected using an anonymous questionnaire. The presence of caries was recorded using the decayed missed filled tooth (DMFT) index, gingival conditions with community periodontal index (CPI) following World Health Organization recommendations. Stepwise logistic regression was applied to determine whether the type of school attended was associated with the clinical outcome variables and subjects' SPOH and with oral health or lifestyle behaviour. RESULTS: The prevalence of caries was 59.8% (95% CI = 41.3 to 76.8); a third of participants showed a CPI = 0, whereas 34.9% had bleeding at probing and 37.9% had calculus. Caries distribution was significantly related to the type of school attended in each score of the DMFT categorisation (P < 0.01). Experience of tooth discomfort was claimed by 23.6% of subjects from grammar schools and 35.3% from technical schools (P < 0.01). In males, CPI > or = 1 was associated with the type of school, OR = 1.6 (95% CI = 1.1 to 2.5), the highest DMFT score and experience of self-perceived tooth discomfort, OR = 1.6 (95% CI = 1.2 to 2.0) and OR = 1.2 (95% CI = 1.4 to 2.6), respectively. In females, only CPI > or = 1 and highest DMFT score were associated with the type of school, OR = 2.1 (95% CI = 1.3 to 3.3) and OR = 1.6 (95% CI = 1.2 to 2.0), respectively. CONCLUSIONS: The type of school is a sensitive indicator of oral health status among adolescents. PMID- 20011750 TI - Relationship between breast- and bottle-feeding and non-nutritive sucking habits. AB - PURPOSE: The objective of the present study was to assess the persistence of non nutritive sucking habits and its relationship with breastfeeding, as well as to establish the influence of sociodemographic factors on these habits among children aged 3 to 5 years. MATERIALS AND METHODS: A case-control study was conducted with 1107 children from public and private daycare centres in Natal, Brazil: 450 in the case group (312 pacifier suckers and 138 thumb suckers) and 657 in the control group (habit-free). Data regarding sociodemographic conditions and duration of breastfeeding were obtained using a structured questionnaire. RESULTS: Breastfeeding for a duration of > 6 months (adjusted odds ratio = 0.311; 95% confidence interval = 0.226 to 0.428) was an independent protective factor against persistent pacifier sucking. The use of pacifiers was more frequent among 3-year-old children and among those from a higher income family and a higher level of schooling of parents. The relation between duration of breastfeeding and thumb sucking was not statistically significant (P = 0.087). There was an association between the thumb sucking habit with sex (female), low level of schooling of father and the child being born as the last male child in the birth order. No relation was found between breastfeeding and habit frequency (day/night and night) (P = 0.301). An association with habit frequency was found with the age of 3 years, female, the family income group that ranged between US $176 and US $875, and those attending private institutions. CONCLUSIONS: Breastfeeding for a duration of > 6 months was a protective factor against the persistence of pacifier sucking, but the subjectivity of the mother-child relationship must be investigated further. PMID- 20011751 TI - Perceived barriers to the provision of preventive care among Iranian dentists. AB - PURPOSE: The aim of this study was to examine the perceived barriers to preventive dental practice among Iranian dentists. MATERIALS AND METHODS: A questionnaire survey was performed among the dentists who participated in two annual dental meetings (conducted in December 2004 and in July 2005) in Tehran, Iran. The dentists who attended the meeting completed the self-administered questionnaire anonymously. The questionnaire included 12 statements with answer alternatives rated on a 5-point scale to which the dentists were asked to indicate how much each of the 12 items, later classified as three barriers (practice-, dentist- and patient-related), impedes them from practising preventive care. The sum of the scores of the four items in each barrier served as an indicator of each dentist's perception of the strength of each barrier. The background data included dentists' personal and professional characteristics. In total, 1033 dentists completed the questionnaire, of whom 980 (64% men) were eligible. Statistical evaluation was performed using analysis of variance, t test and Pearson's correlation coefficient. RESULTS: Of the respondents, 29% to 66% rated the items of the practice-related barrier as very strong impediments. Corresponding figures for the dentist- and the patient-related barriers were 20% to 54% and 70% to 77%, respectively. The highest mean of the score sum was for the patient-related barrier (11.9) and the lowest mean was for the dentist related barrier (7.9). For both practice- and dentist-related barriers, male dentists reported significantly higher scores than the female dentists (P < 0.001). CONCLUSIONS: The perceived barriers to the provision of preventive care should be meticulously explored and tackled to enhance oral health in Iran. PMID- 20011752 TI - The effect of surface roughness of silicone-based resilient liner materials on the adherence of Candida albicans and inhibition of Candida albicans with different disinfectants. AB - PURPOSE: The objective of the present study was to evaluate surface roughness and adherence of Candida albicans and to determine the effectiveness of denture cleansers in the disinfection of silicone-based soft lining materials. MATERIALS AND METHODS: Two types of silicone-based soft lining materials (four room temperature polymerised soft lining materials [RTPSLM]: Ufi Gel P, Ufi Gel C, Mollosil and Soft-Liner; and two high temperature polymerised soft lining materials [HTPSLM]: Moloplast B and Luci Soft) and seven denture cleansers (Efferdent, Polident, Steradent, Corega, Denclen, Klorhex and Axion) were tested in the present study. For each soft lining material, 56 samples (10 mm x 10 mm x 3 mm) were prepared. Surface roughness measurements were made using a profilometer with an optical scanner probe. All samples were ultrasonically cleaned in water for 15 s, autoclave sterilised and contaminated with C. albicans solutions for adherence assay evaluation. After the evaluation, the samples were immersed in seven different denture cleansers. The results were expressed as the ratio of cells remaining after each cleanser in comparison with the control group. RESULTS: HTPSLM showed a lower surface roughness value (2.21 +/- 0.30 lm) than RTPSLM (2.39 +/- 0.40 lm), but these differences were not statistically significant (P > 0.05). Also, HTPSLM showed lower adhesion of C. albicans (mean ranks 136.35) than RTPSLM (mean ranks 138.64), but the differences were not statistically significant (P > 0.05, one- or two-way analysis of variance). In the C. albicans removal test, all denture cleansers showed a strong effect on both silicone-based soft lining materials. HTPSLM showed lower removal activity (mean ranks 30.01) than RTPSLM (mean ranks 25.86), but these differences were not statistically significant (P > 0.05). Although there were no significant differences between disinfectants on both silicone-based soft lining materials, Axion was found to be the most effective disinfectant. Corega, Polident, Denclen, Steradent and Efferdent showed the most effective cleansing, respectively. CONCLUSIONS: RTPSLM and HTPSLM did not show significant differences in the surface roughness value; also they showed no significant differences in the adhesion rate of C. albicans. All tested silicone-based soft lining materials showed adherence of C. albicans after contamination procedures, and removal activity for C. albicans was observed in all tested silicone-based soft lining materials after disinfection procedures. PMID- 20011753 TI - Social, psychological and health concerns among people living with HIV/AIDS in Nigeria. AB - PURPOSE: The aim of the present study was to identify the concerns and the influence of demographic factors on the social, psychological and health concerns of people living with HIV/AIDS (PLWHA) in a Nigerian population. MATERIALS AND METHODS: A questionnaire survey was conducted among 141 consenting PLWHA in the Ife-Ijesa zone, Osun State, Nigeria. A 22-item questionnaire elicited information on PLWHA's concerns about social, psychological and health issues. RESULTS: The main social concern was that of 'Feeling very lonely', whereas the major psychological concern was that of 'Lack of interest in things'. The most common health concern reported was 'Gradually getting thinner', whereas 'No cure for AIDS' was the least. 'Getting tired easily' was also a highly rated concern. Males had statistically significant fewer social concerns than females. Unemployed PLWHA had fewer social, health and psychological concerns when compared with the employed. There was a statistically significant relationship between age and health concerns (P < 0.05). CONCLUSIONS: PLWHA in the present study reported that they were concerned about social, psychological and health issues, despite the fact that they were attending support group meetings. Efforts on care should not only focus on health and medical issues but also on social and psychological issues. PMID- 20011754 TI - Oral health knowledge, attitude and practice among NCC Navy Cadets and their correlation with oral hygiene in south India. AB - PURPOSE: The objective of the present study was to determine oral health knowledge, attitude and practice among National Cadet Corps (NCC) Navy Cadets and their correlation with oral hygiene in South India. MATERIALS AND METHODS: A questionnaire study was conducted among 412 high school students who were voluntary participants of an NCC unit in South India. Knowledge, attitude and practice scores were calculated, and then were correlated with Simplified Oral Hygiene Index (OHI-S) values. Spearman's correlation test was used to measure the correlation. The significance value was set at P < 0.05. Statistical analysis was performed using the Statistical Package for Social Sciences software for Windows, version 13. RESULTS: NCC Cadets' knowledge about gum diseases and the use of fluoride-containing dentifrice was found to be low, dental visits were infrequent and the consumption of sweets was found to be high. There was no correlation between knowledge and attitude scores with that of OHI-S, but a significant correlation was seen between practice and OHI-S scores. CONCLUSIONS: Most of the cadets did not have knowledge about the causes and the prevention of dental diseases. A significant number of participants sought further information, hence it can be said that the cadets had a positive attitude towards oral health. There was no correlation seen between knowledge and attitude scores with that of OHI-S measured, but a significant correlation was noted between practice and OHI-S scores. PMID- 20011755 TI - Smoking and periodontal disease: clinical evidence for an association. AB - PURPOSE: The aim of the present study was to assess the periodontal condition and smoking status, according to dose and duration information, and to estimate the percentage of subjects with periodontitis attributable to cigarette smoking in a representative adult rural population in southern Brazil. MATERIALS AND METHODS: Bivariate statistical analysis was used to evaluate the association of smoking status with periodontitis in a cross-sectional study comprising 165 dentate individuals, aged 35 to 66 years, subjected to oral clinical examination of six sites per tooth in all sextants. RESULTS: The prevalence of periodontitis (having > or = 1 pocket of > or = 4 mm around the index teeth) in the population was 35.2%. Overall, 13.9% had a cumulative loss of attachment > 4 mm; 35.7% of subjects were current smokers, classified as heavy (average 25.3 pack years), moderate (average 14.6 pack years) and light smokers (average 3.1 pack years). Statistical analysis showed that current smokers had an 11 times (95% confidence interval [CI] = 4.69 to 26.62) and former smokers had a nine times (95% CI = 3.29 to 25.96) greater probability of having established periodontitis compared with non-smokers. The number of pack years (P = 0.0004) and years of smoking exposure (P = 0.0013) were associated with an increased prevalence of periodontitis. The number of current smokers with periodontitis might be reduced by 80%, had they not smoked cigarettes. Of the subjects with periodontitis, 64% could be prevented among current smokers by eliminating tobacco consumption. CONCLUSIONS: Cigarette smoking was strongly associated with periodontitis, and there was a relationship with dose and duration of smoking. These findings contributed to the evidence of smoking as a risk factor for periodontal disease and support the importance of dose-response analysis on determining the strength of this association. PMID- 20011756 TI - Restoration margins in young adolescents: a clinical and radiographic study of Swiss Army recruits. AB - PURPOSE: The aim of the present study was to report the radiographical prevalence of overhanging fillings in a group of Swiss Army recruits in 2006 and to relate the dimensions of the overhangs to clinical parameters. MATERIALS AND METHODS: A total of 626 Swiss Army recruits were examined for their periodontal conditions, prevalence of caries, and stomatological and functional aspects of the masticatory system and halitosis. In particular, the present report deals with the presence or the absence of fillings, the presence or the absence of overhangs and their relation to clinical and radiographic parameters. RESULTS: A total of 16,198 interdental sites were evaluated on bitewing radiographs. Of these sites, 15,516 (95.8%) were sound and 682 (4.2%) were filled. Amalgam restorations were found in 94.1% and resin composite fillings in 5.9% of the sites. Of these 682 sites, 96 (14.1%) yielded overhanging margins of various sizes. This low prevalence of fillings represents not only a substantial reduction when compared with a similar Swiss Army study (Lang et al, 1988), but also an improvement in the quality of dental care delivery to young Swiss males. Plaque Index and Gingival Index increased statistically significantly with the presence of fillings, when compared with healthy non-filled sites. Clinical parameters that were significantly associated with the presence of overhangs included clinical attachment loss. Moreover, between 1985 and 2006 the prevalence of fillings was significantly reduced from 20.0% to 4.2% of all surfaces. Furthermore, the marginal fit of the fillings improved from 33.0% with overhangs to 14.1%. CONCLUSIONS: A significant improvement was observed in the periodontal and dental conditions of young Swiss males that was shown to have taken place within the previous two decades. From 1985 to 2006, the prevalence of fillings was reduced fourfold and that of overhanging margins twofold, documenting an improvement in the quality of restorative dentistry. PMID- 20011758 TI - Patients' attitudes towards oral health and experiences of periodontal treatment: a qualitative interview study. AB - PURPOSE: The aim of the present qualitative study was to analyse patients' attitudes towards oral health and experiences of periodontal treatment. MATERIALS AND METHODS: Patients who were referred to a specialist clinic in periodontics were subjected to open-ended interviews before (Abrahamsson et al, 2008), as well as after, the phase of cause-related periodontal therapy. An interview guide was used with themes of interest. The time interval between the interviews was about 6 months. Because the study focused on psychosocial interactions, the qualitative method used for the analysis was Grounded Theory (Glaser and Strauss, 1967). Data collection and analysis were simultaneous processes that focus on psychosocial processes, existing problems and how these problems were dealt with. The interviews were tape-recorded and transcribed verbatim, and consequently analysed in a hierarchical coding process until saturation was met (n = 16). RESULTS: In the analysis, a core category was identified: 'Understanding the seriousness of the disease condition'. This core concept was related to four additional categories and dimensions: 'The need to be treated respectfully', 'To gain insight', 'Frustration about the financial cost for treatment' and 'Feelings of control over the situation'. Taken together, the core category and its related subcategories described a psychosocial process that was related to periodontal treatment. CONCLUSIONS: The results elucidated the patients' vulnerability and the importance of patient communication in the treatment for chronic periodontitis in order for the patient to feel in control of the situation. PMID- 20011757 TI - Clinical effects of interdental cleansing on supragingival biofilm formation and development of experimental gingivitis. AB - PURPOSE: The aim of the present study was to test the effects of interdental cleansing with dental floss on supragingival biofilm removal in natural dentition during a 3-week period of experimental biofilm accumulation. MATERIALS AND METHODS: The present study was performed as a single-blind, parallel, randomised, controlled clinical trial using the experimental gingivitis model (Loe et al, 1965). Thirty-two students were recruited and assigned to one of the following experimental or control groups: Group A used a fluoride-containing dentifrice (NaF dentifrice) on a toothbrush for 60 s twice a day, Group B used an unwaxed dental floss twice a day, Group C used a waxed dental floss twice a day in every interproximal space and Group D rinsed twice a day for 60 s with drinking water (control). RESULTS: During 21 days of abolished oral hygiene, the groups developed various amounts of plaque and gingivitis. Neither of the cleansing protocols alone allowed the prevention of gingivitis development. Toothbrushing alone yielded better outcomes than did any of the flossing protocols. Interdental cleansing with a waxed floss had better biofilm removal effects than with unwaxed floss. CONCLUSIONS: Toothbrushing without interdental cleansing using dental floss and interdental cleansing alone cannot prevent the development of gingivitis. PMID- 20011759 TI - Untreated dental caries in a Brazilian paediatric AIDS patient population. AB - PURPOSE: The objective of the present study was to assess the prevalence of untreated caries in a Brazilian paediatric acquired immunodeficiency syndrome (AIDS) patient population and its association with sociodemographic, behavioural and clinical characteristics. MATERIALS AND METHODS: The study group was comprised of 125 HIV-infected patients (aged 3 to 15 years) who had already manifested AIDS and were assisted in a specialised health care unit. Dental examinations followed the World Health Organization's guidelines for oral health surveys. Family caregivers provided information about the socioeconomic standing and the behaviour of their children. Patients' medical records in the hospital provided information on the clinical status of patients. A Poisson regression analysis was used for assessing the covariates for the prevalence of untreated dental caries, as adjusted by age. RESULTS: The prevalence of untreated caries was 58%; a higher prevalence was found in younger children with primary and mixed dentition. The prevalence of untreated caries associated significantly with lower socioeconomic status (household crowding and schooling of the caregiver), dietary habits (higher frequency of sugar consumption) and poorer clinical status (HIV viral load and symptom severity). CONCLUSIONS: The high burden of untreated caries on paediatric AIDS patients reinforced the importance of integrating the clinician with the interdisciplinary health care team that assisted these children. The identification of socioeconomic and behavioural factors associated with caries experience reinforced the importance of the attention that children with AIDS received within their own households for the prevention of dental disease, particularly a proper nutritional advisement and monitoring of dental hygiene. PMID- 20011761 TI - The importance of knowledge. PMID- 20011762 TI - Laser-assisted cavity preparation and adhesion to erbium-lased tooth structure: part 1. Laser-assisted cavity preparation. AB - The use of the ruby laser (693.4 nm) was first described in 1960, and it was applied for hard tissue ablation in 1964. Different wavelengths [Nd:YAG (1.065 microm), CO2 (9.6 microm), Ho:YAG (2.12 microm)] were consequently explored. Due to massive thermal side effects, these wavelengths caused increased temperature in dental pulp, as well as microcracks and carbonization. The use of this laser for dental hard tissue preparation was eventually abandoned. At the end of the 1980s, excimer lasers (ultraviolet) and the erbium laser (infrared) were developed, with the advantages of improved temperature control and smaller penetration depths. With the development of smaller devices and improved knowledge of how to limit damage to the surrounding tissues, new ablation techniques were established in the 1990s. There is still contradiction in the current literature, however, in that different wavelengths are advocated for hard tissue removal, and heterogeneity in laser parameters and power densities remain. In this review, the effects of the wavelengths presently used for cavity preparation are evaluated. We conclude that erbium lasers (Er:YAG and Er,Cr:YSGG) are most efficient and, with the right parameters, the thermal side effects are small. There is a substantial need for "gold standards", although this is difficult to establish in practice owing to different laser parameters (including pulse repetition rate, amount of cooling, energy delivered per pulse, and types of pulses) and target specificity (tissue interaction with sound or decayed enamel or dentin, and the extent of (de)mineralization) which influence tissue interaction. PMID- 20011763 TI - Information resources and material selection in bonded restorations among Korean dentists. AB - PURPOSE: To elucidate how dentists access knowledge sources when choosing adhesive agents for bonded restoration and whether these resources influenced the selection of materials. MATERIALS AND METHODS: A national survey was carried out involving dentists in South Korea. The questionnaire included the status of the operator, clinical techniques, and materials. A total of 12,193 e-mails were distributed, 2632 were opened by recipients, and 840 responses were collected. RESULTS: For primary information resources in material selection, 55.8% (n = 469) of the responders mentioned continuing education. This selection criteria varied according to the working status of the clinician (p < 0.0001). The choice of bonding system was influenced by of the length of the career (p < 0.0001), and the preferred bonding systems differed according to the source of the information (p = 0.035). CONCLUSION: The popular bonding systems were not ranked according to the bonding strategy, but a preference was found for specific brands or manufacturers. To decrease the confusion of clinicians related to the selection of bonding materials, evidence-based guidelines need to be comprehensibly organized and efficiently approached in clinics. PMID- 20011764 TI - Preliminary evaluation of titanium tetrafluoride as an alternative ceramic etchant to hydrofluoric acid. AB - PURPOSE: The etching of dental ceramics with a glassy matrix by means of hydrofluoric acid (HF) is highly effective and after the application of a silane coupling agent, a strong link between the adhesive and the ceramic is achieved. The purpose of this study was to investigate whether aqueous titanium tetrafluoride (TiF4) solution could be used as an alternative ceramic etchant or not. MATERIALS AND METHODS: Seventy lithia-based all-ceramic specimens (IPS Empress 2, Ivoclar Vivadent) (diameter: 4 mm, height: 4 mm) were prepared and ultrasonically cleaned for 15 min in ethanol and deionized water, then divided into 1 control and 4 experimental groups (n = 14/group). The control group was acid etched with 4.9% HF for 20 s as described by the manufacturer. The experimental groups were treated with 2.5% TiF4 solution for 60 s, 2.5% TiF4 solution for 120 s and 5% TiF4 solution for 60 s, as well as 5% TiF4 solution for 120 s. Seven of the ceramic specimens in each group were luted to the other seven by a dual-curing cement (Variolink II, Ivoclar Vivadent,) with silane application (Monobond-S). After storage of luted specimens in deionized water at 37 degrees C for 24 h, the specimens were embedded in plastic holders with cyanoacrylate, and the shear-bond strength (SBS) tests were performed with a Shimadzu universal testing machine at a crosshead speed of 0.5 mm/min. A one-way ANOVA and Bonferroni correction, followed by the Tukey test, were applied for statistical analysis. Scanning electron microscopic examination was performed to evaluate the pattern of debonding. RESULTS: Significantly higher SBS values (p 0.05). CONCLUSION: Aqueous TiF4 solution exhibited similar shear bond strength values in comparison to commonly used HF and can be considered as an alternative ceramic etchant within the limitations of this preliminary report. PMID- 20011765 TI - Phosphoric acid/sodium hypochlorite mixture as dentin conditioner: a new approach. AB - PURPOSE: To investigate the effect of a phosphoric acid/sodium hypochlorite (NaOCl) mixture as a dentin conditioner on shear bond strength, the resin/dentin interface, as well as the surface area and tubular diameter of dentin. MATERIALS AND METHODS: Thirty human molars were prepared for shear bond strength testing. The teeth were randomly divided into five groups according to the method of dentin conditioning. In group I, dentin was conditioned with 37% phosphoric acid, in group II dentin was conditioned with 37% phosphoric acid followed by application of 5.25% NaOCl, and in groups III, IV, and V, dentin was conditioned with a phosphoric acid/NaOCl mixture for 15, 30, and 60 s, respectively. A resin based restorative composite disk was bonded to the occlusal surface of each tooth and shear bond strength was measured in a universal testing machine. The resin/dentin interface was evaluated using a scanning electron microscope (SEM). The surface area and tubular diameter for different conditioning methods were assessed using atomic force microscopy (AFM). The data were submitted to one-way ANOVA at a significance level of 0.05 followed by the Tukey-Kramer Multiple comparisons test. RESULTS: Group III showed the highest shear bond strength value (36 MPa), which was significantly higher than groups I and II. SEM micrographs revealed hybrid layer formation for all groups except group II. AFM results showed that groups II and V have significantly higher surface areas and dentinal tubular diameters than other groups. CONCLUSION: The use of a phosphoric acid/NaOCl mixture as dentin conditioner improved the bond strength of an etch and-rinse single-bottle adhesive to dentin. PMID- 20011766 TI - Demineralization effects of hydrogen peroxide on bovine enamel and relation to shear bond strength of brackets. AB - PURPOSE: To measure the demineralization capacity of 37% phosphoric acid and shear bond strength (SBS) of brackets attached on bovine enamel at different times after bleaching with 30% hydrogen peroxide. MATERIALS AND METHODS: Four equally-sized pieces of each crown from 18 bovine incisors were randomly distributed among 7 groups (n = 10). After bleaching with 30% hydrogen peroxide for 1 h, specimens were stored in artificial saliva for 0 h, 24 h, or 1, 2, 3, or 4 weeks before bonding specimens to brackets. An unbleached group of specimens served as controls. Shear bond strength (SBS in MPa) was measured with a universal testing machine. Adhesive Remnant Index (ARI) scores were determined after failure of bracket bonds. To measure demineralized Ca2+, four 4 x 4 mm sections from each of 15 bovine incisors were randomly distributed among 4 groups (n = 15). Specimens were stored in artificial saliva for 0 h, 24 h or 7 days after bleaching and then immersed in 37% phosphoric acid solution. After 15 s, 30 s, and 60 s, 5-ml aliquots of solution were removed for spectrophotometry. Unbleached specimens served as controls. RESULTS: Larger amounts of Ca2+ were extracted from the enamel by phosphoric acid up to 24 h after application of hydrogen peroxide, when there was also a significant decrease in bracket-enamel SBS. After 1 week, there were no changes in amounts of Ca2+ extracted, and SBS values returned towards unbleached values. CONCLUSION: Lower bracket-enamel SBS values at 24 h after bleaching are closely correlated with the larger amounts of Ca2+ extracted from the enamel. PMID- 20011767 TI - The effect of water storage, elapsed time and contaminants on the bond strength and interfacial polymerization of a nanohybrid composite. AB - PURPOSE: To systematically characterize the effect of time lapse, water storage, and selected contaminants on the bond strength of a nanofilled dental composite. MATERIALS AND METHODS: Half-dumbbell-shaped samples were fabricated out of light polymerizing composite resin. To function as substrates they were aged for 30 days in water. Prior to bonding, the substrates' surfaces were subjected to the following treatments: 1) Removing a 0.2- to 0.4-mm layer using a fluted carbide bur; 2) grit blasting with 50 microm alumina particles; 3) etching with phosphoric acid gel; 4) grit blasting followed by etching; 5) blasting with tribochemical particles followed by silane application; 6) sanding with 400-grit paper, air aging of the adherent half-sample before bonding; 7) surface contamination with saliva; 8) surface contamination with blood. In each group (n = 30), freshly polymerized (except in group 6) adherent half-samples were bonded to the substrate half-samples by a layer of unfilled adhesive resin. Fifteen full dumbbell-shaped specimens were subjected to tensile testing after 1 h and 15 after 7 days water storage. In a positive control group, freshly cured half samples were bonded shortly after fabrication. The tensile strength was analyzed using Weibull statistics and presented in terms of the material's characteristic strength and shape parameter. Fractographs of the two weakest and strongest samples of each group were produced. The surfaces were searched to locate hackle, wake hackle and the origin of the fracture. RESULTS: Surface roughness and time lapse increased the bond strength of the repaired specimens. All groups in which surface roughness was produced before bonding increased in repair strength. Post bonding aging improved strength. Fractographs yielded interpretable data whenever larger surfaces of single phase bonding resin were present. CONCLUSION: 1) Roughening and etching an aged composite's surface prior to applying a coat of unfilled resin and the filled material increases repair bond strength by up to 100%. 2) The repair bond strength of a roughened aged composite is 25% to 30% inferior to the tensile strength of solid specimens. 3) After 7 days' storage in water, no detrimental effect could be seen from saliva or blood contamination if the surfaces were properly rinsed. PMID- 20011768 TI - Durability of resin bonding to zirconia ceramic using different primers. AB - PURPOSE: The purpose of this in vitro study was the evaluation of the bond strength and its durability to zirconia ceramic after using two phosphate monomer containing primers. MATERIALS AND METHODS: Plexiglas tubes filled with composite resin were bonded to zirconia ceramic disks (Cercon) which were air abraded (50 microm Al2O3 at 2.5 bar pressure) and ultrasonically cleaned in isopropanol for 3 min. Groups of 16 specimens each were bonded with Multilink Automix using either no primer (NO) or using a metal primer (AP: Alloy Primer, Kuraray) or a metal and zirconia primer (MZ: Metal/Zirconia Primer, Ivoclar Vivadent). Another group bonded with a phosphate monomer (MDP) containing resin (PF: Panavia F, Kuraray) served as control for an established bonding method to zirconia ceramic. Prior to tensile bond strength testing, subgroups of 8 bonded specimens were stored in distilled water (37 degrees C) for either 3 or 150 days. Additionally, the 150 day specimens were thermocycled (TC) between 5 degrees and 55 degrees 37,500 times. Statistical analyses were conducted with the Wilcoxon rank sum test. RESULTS: Without primer, the median initial bond strength of Multilink Automix to zirconia ceramic was relatively low (12.4 MPa). The primers improved the bond strength significantly (p 1 was considered indicative of a diagnosis of OSAS. RESULTS: The prevalence of OSAS was 10.6%. We found a negative correlation between mean annual hemoglobin level and total sleep time with SpO2 < 90% (r = micro0.343; p = 0.002), as well as between mean annual hemoglobin level and total sleep time with SpO2 < 80% (r = micro0.270; p = 0.016). There was no association between AHI and painful crisis. CONCLUSIONS: The prevalence of OSAS in this population was high (10.6%). Therefore, it is important to identify signs of OSAS as soon as possible and to determine the mean annual hemoglobin level because of the inverse correlation between that level and the total sleep time with SpO2 < 90% or < 80%. PMID- 20011843 TI - Ventilator-associated pneumonia: epidemiology and impact on the clinical evolution of ICU patients. AB - OBJECTIVE: Although ventilator-associated pneumonia (VAP) is a major cause of nosocomial infection, its role in the prognosis of patients remains undefined. The objective of this study was to evaluate the impact of VAP on the clinical evolution of patients. METHODS: This was a prospective cohort study involving 233 patients on mechanical ventilation (VAP group, n = 64; control group, n = 169). Primary outcomes were time on mechanical ventilation (TMV), time in ICU (TICU), overall length of hospital stay (LHS) and in-ICU mortality. Secondary outcomes were in-hospital mortality, microbiological profile, prior use of antibiotics and risk factors for VAP acquisition. RESULTS: Control and VAP group outcomes were, respectively, as follows: median TMV (days), 9 (interquartile range [IQR]: 5-15) and 23 (IQR: 15-37; p < 0.0001); median TICU (days), 12 (IQR: 8-21) and 27 (IQR: 17-42; p < 0.0001); median LHS (days), 33 (IQR: 18-64) and 46 (IQR: 25-90; p = 0.05); and in-ICU mortality, 38% (95% CI: 31-45) and 55% (95% CI: 42-67; p = 0.02). VAP was a predictor of in-ICU mortality (OR = 3.40; 95% CI: 1.54-7.48). TMV (OR = 2.27; 95% CI: 1.05-4.87) and prior use of antibiotics (OR = 1.07; 95% CI: 1.04-1.10) were risk factors for VAP. VAP did not affect in-hospital mortality. Acinetobacter spp. was the most common isolate (28%). Inappropriate empirical antibiotic therapy was administered in 48% of cases. CONCLUSIONS: In this study, there was a high incidence of infection with resistant bacteria and inappropriate initial antibiotic therapy. Long TMV and prior use of antibiotics are risk factors for VAP. PMID- 20011844 TI - Bacteriological analysis of induced sputum for the diagnosis of pulmonary tuberculosis in the clinical practice of a general tertiary hospital. AB - OBJECTIVE: To determine the diagnostic sensitivity of bacteriological analyses in induced sputum (IS) for the diagnosis of pulmonary tuberculosis (TB) and to identify the clinical characteristics associated with the confirmed diagnosis, as well as to determine the diagnostic yield of bronchoscopy carried out when IS tests negative for AFB in smear microscopy. METHODS: A retrospective, cross sectional study of patients suspected of having active pulmonary TB and referred to our clinic for sputum induction. We consecutively reviewed the laboratory data of all patients submitted to sputum induction between June of 2003 and January of 2006, as well as their electronic medical records. In addition, the results of the bacteriological analysis of bronchoscopic specimens collected from the patients whose AFB tests were negative in IS were reviewed. RESULTS: Of the 417 patients included in the study, 83 (19.9%) presented IS samples that tested positive for TB (smear microscopy or culture). In the logistic regression analysis, radiological findings of cavitation (OR = 3.8; 95% CI: 1.9-7.6) and of miliary infiltrate (OR = 3.7; 95% CI: 1.6-8.6) showed the strongest association with the diagnosis of pulmonary TB. In 134 patients, bronchoscopy was carried out after negative AFB results in IS and added 25 (64.1%) confirmed diagnoses of pulmonary TB. CONCLUSIONS: In our clinical practice, the frequency of confirmed diagnosis of pulmonary TB using IS (19.9%) was lower than that previously reported in controlled trials. Cavitation and miliary infiltrate increase the diagnostic probability of pulmonary TB using IS. The use of bronchoscopy when IS tests negative for AFB significantly increases sensitivity in the diagnosis of pulmonary TB. PMID- 20011845 TI - Tuberculosis treatment: integration between hospitals and public health care clinics in the city of Sao Paulo, Brazil. AB - OBJECTIVE: To evaluate the level of access to health care clinics of a population of patients hospitalized for tuberculosis (TB) at two hospitals in the city of Sao Paulo, Brazil, comparing them with a population of TB patients under outpatient treatment only. METHODS: We compiled sociodemographic, clinical and epidemiological data related to patients hospitalized for TB at two hospitals in the city of Sao Paulo, Brazil, between January and December of 2007, using a structured questionnaire. We also identified the outpatient clinics to which the patients were referred at discharge. The same variables were evaluated for TB outpatients during the same period, using a database. RESULTS: The study sample consisted of 474 patients (166 inpatients and 308 outpatients: mean age, 41.0 and 39.1 years, respectively). The univariate analysis showed positive associations between hospitalization due to TB and the following variables: 30-39 year age bracket (OR = 2.17); 50-59 year age bracket (OR = 2.17); combination of pulmonary and extrapulmonary forms of TB (OR = 5.31); TB re-treatment (OR = 2.66); seeking treatment at other health care facilities prior to the diagnosis of TB (OR = 2.05); symptom duration of more than 12 weeks (OR = 2.23); and TB diagnosed at hospitals or in emergency rooms (OR = 4.68). The proportion of inpatients who resided in the same regional health district area as that in which the respective hospital was located was 77.6% and 36.8%. The proportion of discharged patients who were referred to outpatient clinics in the same areas was 67.1% and 39.7%, respectively. CONCLUSIONS: Patients hospitalized for TB should be monitored from discharge until their admission to the outpatient clinic. PMID- 20011846 TI - Ex vivo lung perfusion: initial Brazilian experience. AB - In the last 20 years, lung transplantation has become the standard treatment for patients with end-stage lung disease. However, less than 20% of the donor lungs available for transplant are actually usable. This disparity between the growing number of recipients and the small number of donors has resulted in increased mortality among lung transplant candidates on waiting lists. Strategies such as the utilization of organs from marginal donors have proven ineffective in increasing the number of transplants. In 2000, a new method for reconditioning human lungs that had been previously rejected for transplantation was developed in Sweden. We describe our initial experience with ex vivo lung perfusion. PMID- 20011847 TI - Pulmonary rehabilitation in COPD: from exercise training to 'real life'. AB - The objective of this study was to assess the efficacy of exercise training by means of a rehabilitative walking protocol. Twenty patients with COPD staged as III/IV according to the Global Initiative for Chronic Obstructive Lung Disease were included. Patients were evaluated at baseline and at the end of the exercise program regarding oxygen desaturation, sensation of dyspnea/fatigue, quality of life and six-minute walk distance. The comparison between the six-minute walk distance prior to and after training proved the efficacy of the protocol, which also resulted in a decrease in oxygen desaturation and in the sensation of dyspnea. This protocol can be easily put into practice. PMID- 20011848 TI - Nosocomial pneumonia: importance of the oral environment. AB - Nosocomial pneumonia, especially ventilator-associated pneumonia, is a common infection in ICUs. The main etiologic factors involve colonizing and opportunistic bacteria from the oral cavity. Oral hygiene measures, including the use of oral antiseptic agents, such as chlorhexidine, have proven useful in reducing its incidence. The objective of this article was to review the literature on the importance of the oral environment in the development of nosocomial pneumonia. PMID- 20011849 TI - Methods for the assessment of peripheral muscle fatigue and its energy and metabolic determinants in COPD. AB - It has been well established that, in addition to the pulmonary involvement, COPD has systemic consequences that can lead to peripheral muscle dysfunction, with greater muscle fatigue, lower exercise tolerance and lower survival in these patients. In view of the negative repercussions of early muscle fatigue in COPD, the objective of this review was to discuss the principal findings in the literature on the metabolic and bioenergy determinants of muscle fatigue, its functional repercussions, as well as the methods for its identification and quantification. The anatomical and functional substrate of higher muscle fatigue in COPD appears to include lower levels of high-energy phosphates, lower mitochondrial density, early lactacidemia, higher serum ammonia and reduced muscle perfusion. These alterations can be revealed by contraction failure, decreased firing rates of motor units and increased recruitment of motor units in a given activity, which can be functionally detected by a reduction in muscle strength, power and endurance. This review article also shows that various types of muscle contraction regimens and protocols have been used in order to detect muscle fatigue in this population. With this understanding, rehabilitation strategies can be developed in order to improve the resistance to muscle fatigue in this population. PMID- 20011850 TI - Chapter 3--pulmonary cryptococcosis. AB - Cryptococcosis is a systemic mycosis caused by two species of the encapsulated basidiomycetes, Cryptococcus neoformans and C. gattii, which, respectively, cause infection in immunocompromised individuals and in immunologically normal hosts. Patients with T-cell deficiencies are more susceptible to this infection. The spectrum of the disease ranges from asymptomatic pulmonary lesions to disseminated infection with meningoencephalitis. The medical relevance of cryptococcosis increased dramatically as a consequence of the AIDS epidemic and organ transplants. PMID- 20011851 TI - Chapter 4--histoplasmosis. AB - Histoplasmosis is systemic mycosis caused by a small fungus, Histoplasma capsulatum var. capsulatum, whose natural habitat is soil contaminated by bat or bird excrement. The incidence of histoplasmosis is worldwide. In Brazil, the disease is found in all regions; however, the state of Rio de Janeiro is responsible for most of the microepidemics described. Human infection occurs when airborne spores of H. capsulatum are inhaled. The most common clinical presentation is asymptomatic. The symptoms of acute or epidemic histoplasmosis are high fever, cough, asthenia and retrosternal pain, as well as enlargement of the cervical lymph nodes, liver and spleen. The most common radiological findings are diffuse reticulonodular infiltrates in both lungs, as well as hilar and mediastinal lymph node enlargement. In chronic pulmonary histoplasmosis, the clinical and radiological manifestations are identical to those of reinfection with pulmonary tuberculosis. Histoplasmosis is diagnosed by means of the identification or culture growth of the fungus in sputum or fiberoptic bronchoscopy specimens. Histopathological examination reveals the fungus itself within or surrounding macrophages, as well as granulomatous lesions with or without caseous necrosis. Double agar gel immunodiffusion is the most easily used and readily available serologic test for making the immunological diagnosis. Acute histoplasmosis with prolonged symptoms requires treatment, as do the disseminated or chronic pulmonary forms of the disease. The drug of choice is itraconazole. PMID- 20011852 TI - A rare case of co-infection with pulmonary tuberculosis and oronasal actinomycosis. AB - Oronasal actinomycosis is an infection seldom described in the literature, especially in the form of a co-infection with pulmonary tuberculosis. We report the case of a 48-year-old male admitted to the isolation ward due to active pulmonary tuberculosis, with a history of diabetes and alcohol abuse. While hospitalized, the patient complained of dysphagia and nasal regurgitation of food. The examination of the oral cavity revealed an oronasal fistula. The infecting agent was identified, and the treatment was successful. We also present a brief review of the literature, as well as a full description and discussion of the process of investigating this rare clinical case. PMID- 20011853 TI - Endoscopic treatment of tracheobronchial tree fistulas using atrial septal defect occluders: preliminary results. AB - Fistulas in the tracheobronchial tree (bronchopleural and tracheoesophageal fistulas) have a multifactorial etiology and present a variable incidence in the literature. In general, the related morbidity and mortality are high. Once such a fistula has been diagnosed, surgical closure is formally indicated. However, the clinical status of affected patients is usually unfavorable, which precludes the use of additional, extensive surgical interventions. In addition, attempts at endoscopic closure of these fistulas have seldom been successful, especially when the fistula is large in diameter. We report the cases of three patients submitted to endoscopic closure of fistulas, two of which were larger than 10 mm in diameter, by means of the insertion of atrial septal defect occluders. The procedure was minimally invasive, and the initial results were positive. The results indicate that this is a promising technique for the resolution of tracheobronchial tree fistulas. PMID- 20011854 TI - Chronic cavitary pulmonary histoplasmosis. AB - Histoplasmosis is a systemic mycosis caused by the thermally dimorphic fungus Histoplasma capsulatum, which can be isolated from soil contaminated with droppings from birds or bats. Chronic cavitary pulmonary histoplasmosis is one of the rarest clinical presentations of this disease. The differential diagnosis with tuberculosis should be made in patients presenting with cavitated lesions in upper lung segments. We report the case of a female patient with chronic cavitary pulmonary histoplasmosis who had presented with progressive dyspnea and worsening of the radiological pattern over a four-year period. PMID- 20011855 TI - Epidural lumbar block or lumbar plexus block combined with general anesthesia: efficacy and hemodynamic effects on total hip arthroplasty. AB - BACKGROUND AND OBJECTIVES: Anesthesia for total hip arthroplasty (THA) is a challenge due to the advanced age and associated diseases of patients. The objective of this study was to evaluate whether the efficacy of the nociceptive blockade, secondary hemodynamic effects, difficulty to execute the technique, and influence in intraoperative bleeding of lumbar plexus block combined with general anesthesia is equivalent to epidural lumbar block in patients undergoing THA. METHODS: Patients with physical status ASA I to III were randomly separated into two groups, Epidural and Lumbar. In the Epidural group, continuous epidural lumbar block with 10 to 15 mL of 0.5% ropivacaine was performed. Patients in the Lumbar group underwent posterior lumbar plexus block with 0.4 mL.kg(-1) of 0.5% ropivacaine. All patients underwent general anesthesia. The difficulty to perform the technique, its efficacy, and secondary hemodynamic effects were evaluated. RESULTS: Forty-one patients were included in this study. The length of time to execute the epidural block was shorter, but the number of attempts to position the needle was similar in both groups. Epidural block was more effective. In the Lumbar group, an increase in diastolic blood pressure and mean arterial pressure (MAP) and in the double product was observed after the incision, and anesthetic consumption was greater. After the blockade, MAP was lower 50, 60, and 70 minutes after the epidural block. Bleeding was similar in both groups. CONCLUSIONS: Nociceptive blockade, which was not associated with hemodynamic instability when combined with general anesthesia, was more effective in epidural block. Lumbar plexus block proved to be a useful technique when combined with general anesthesia when epidural block is contraindicated. PMID- 20011856 TI - Supraclavicular brachial plexus block: a comparative clinical study between bupivacaine and levobupivacaine. AB - BACKGROUND AND OBJECTIVES: Brachial plexus block is used in surgical procedures of the upper limbs. The brachial plexus is a potential territory for absorption of local anesthetics. Studies on bupivacaine isomers have shown reduced cardiovascular toxicity of its levorotatory form (levobupivacaine). However, the anesthetic efficacy (sensorial and motor blockades) of levobupivacaine in neuroaxis blocks has been debated. The objective of this study was to demonstrate the anesthetic efficacy of levobupivacaine in brachial plexus block, using the perivascular subclavian approach, by comparing it to racemic bupivacaine. METHODS: Fifty adult patients of both genders, ASA I and II, underwent subclavian perivascular brachial plexus block, with the aid of a peripheral neurostimulator, for orthopedic surgeries of the upper limbs. Patients were randomly divided into two groups: G BUPI - racemic bupivacaine, and G LEVO - levobupivacaine, and they received 30 mL of the 0.5% solution. Sensorial blockade was evaluated by pinprick stimulation from C5 to C8 metameres; and the motor blockade was assessed 1, 2, 5, 10, 15, 20, 25, and 30 minutes after the administration of the local anesthetic or until blockade of fingers, hand, forearm, and arm movements was observed. RESULTS: Statistical differences in latency, failure rate, and degree of the motor blockade, and failure of the sensorial blockade between both groups were not observed, but the latency of the sensorial blockade in all metameres analyzed showed statistically significant differences. Adverse events inherent to the administration of local anesthetics were not observed. CONCLUSIONS: The anesthetic efficacy of levobupivacaine in brachial plexus block was similar to that of the racemic solution commonly used. PMID- 20011857 TI - Dose requirement of intrathecal bupivacaine for cesarean delivery is similar in obese and normal weight women. AB - BACKGROUND AND OBJECTIVES: The effect of BMI on the spread of intrathecal bupivacaine is controversial. This study assessed the ED95 of intrathecal bupivacaine for elective cesarean delivery in obese and normal weight women. METHODS: We studied normal weight (BMI < 25 kg x m(-2)) and obese (BMI > 30 kg x m(-2)) women with singleton term pregnancies undergoing elective cesarean delivery. The study was conducted as a single blinded, up-down sequential allocation study (modified by the Narayana rule). All patients received a combined spinal-epidural anesthesia with a variable intrathecal dose of hyperbaric 0.75% bupivacaine, plus fentanyl 10 microg and morphine 100 microg. The first patient received 9 mg of bupivacaine. Supplemental anesthesia was provided through the epidural catheter if required. The primary outcome was successful analgesia, defined as a sensory block to at least T6, and no request for supplemental anesthesia. The ED95 for the satisfactory outcome was determined by a logistic model with non-log-transformed doses. RESULTS: Twenty-four normal weight and sixteen obese patients were enrolled. The estimated ED95 for all forty patients was 12.92 mg (95% CI: 11.49 to 34.77). The estimated ED95 for the normal weight and the obese subgroups were similar at 12.78 mg (95% CI: 10.75 to + infinity) and 11.86 mg (95%CI: 11.31 to 15.61), respectively. CONCLUSIONS: If single shot spinal anesthesia is used for cesarean delivery, obese and normal weight patients should receive similar doses of hyperbaric bupivacaine. Although in our study the effective dose 95% could not be precisely determined, it is possible to state that it is at least 11.49 mg. PMID- 20011858 TI - Postoperative analgesia for the surgical correction of congenital clubfoot: comparison between peripheral nerve block and caudal epidural block. AB - BACKGROUND AND OBJECTIVES: Correction of congenital clubfoot (CCF) is associated with severe postoperative pain. Caudal epidural block associated with general anesthesia is the anesthetic technique used more often in children, but it is limited by the short duration of the postoperative analgesia. Peripheral nerve blocks are associated with a low incidence of complications and prolonged analgesia. The objective of this study was to compare the duration of analgesia in peripheral nerve blocks and caudal block, as well as morphine consumption in the first 24 hours after correction of CCF in children. METHODS: This is a randomized, double-blind study with children undergoing surgeries for posteromedial release of CCF, who were divided in four groups according to the anesthetic technique: caudal (ACa), sciatic and femoral block (IF), sciatic and saphenous block (IS), and sciatic block and local anesthesia (IL), associated with general anesthesia. In the first 24 hours, patients received oral dypirone and acetaminophen, and they were evaluated by anesthesiologists who were unaware of the technique used. Oral morphine (0.19 mg x kg(1) per day) was administered according to the scores of the CHIPPS (Children's and infants' postoperative pain scale) scale. RESULTS: One hundred and eighteen children separated into four groups: ACa (30), IF (32), IS (28), and IL (28) participated in this study. The mean time between the blockade and the first dose of morphine was 6.16 hours in group ACa, 7.05 hours in group IF, 7.58 in IS, and 8.18 hours in IL. Morphine consumption was 0.3 mg.kg-1 per day in all four groups. Significant differences were not observed among the groups. CONCLUSIONS: peripheral nerve blocks did not promote longer lasting analgesia or a decrease in morphine consumption in the first 24 hours in children undergoing CCF correction when compared to caudal epidural block. PMID- 20011859 TI - Clonidine as pre-anesthetic medication in cataract extraction: comparison between 100 microg and 200 microg. AB - BACKGROUND AND OBJECTIVES: The objective of the present study was to evaluate the degree of sedation, intraocular pressure, and hemodynamic changes with premedication with low doses of oral clonidine, 100 microg and 200 microg, in outpatient cataract surgeries. METHODS: This is a randomized, double-blind, clinical study undertaken at the Universidade Federal de Sao Paulo with 60 patients of both genders, physical status ASA 1 and 2, ages 18 to 80 years. Patients were separated into three groups: placebo, clonidine 100 microg, and clonidine 200 microg. Intraocular pressure, heart rate, and blood pressure besides assessment of sedation were measured before and 90 minutes after the administration of clonidine. Sedation levels were classified according to the Ramsay sedation scale. RESULTS: Patients who received placebo and 100 microg of clonidine did not show reduction in heart rate, while a reduction in heart rate was observed in patients who received 200 microg of clonidine, and this difference was statistically significant. Patients who received 200 microg of clonidine also had a reduction in systolic and diastolic blood pressure (p < 0.05). One patient who received 200 microg of clonidine developed severe hypotension, with systolic pressure < 80 mmHg. Patients treated with clonidine had a reduction in intraocular pressure (p < 0.05). Ninety minutes after the oral administration of placebo and 100 microg and 200 microg of clonidine, 25%, 60%, and 80% of the patients respectively were classified as Ramsay 3 or 4. CONCLUSIONS: Clonidine 100 microg can be indicated as premedication for fasciectomies, being effective in sedation and reduction of intraocular pressure, without adverse effects on blood pressure and heart rate. PMID- 20011860 TI - Influence of angiotensin-converting enzyme inhibitors on hypotension after anesthetic induction: is the preoperative discontinuation of this drug necessary? AB - BACKGROUND AND OBJECTIVES: The discontinuation of drugs such as angiotensin converting enzyme inhibitors (ACE inhibitors) has been suggested based on reports of hypotension during anesthesia. This may imply on a higher risk of intraoperative hypertensive peaks with deleterious consequences for the patient. The objective of the present study was to evaluate the influence of the preoperative use of ACE inhibitors on the development of hypotension during anesthesia. METHODS: This is a case-controlled study of patients who developed hypotension after anesthetic induction. The control group was composed of patients of the same age and gender who underwent surgeries during the same period and who did not develop hypotension. Parameters of interest included: age, gender, size of the surgery, prior diagnosis of hypertension, use of ACE inhibitors, physical status (ASA), intraoperative bleeding, anesthetic technique, and duration of the surgery. RESULTS: In 2,179 surgeries, 40 patients developed hypotension. Twenty of those patients used ACEIs on the day of the surgery. The control group was composed by 171 patients, 11 of which used ACE inhibitors. Univariate analysis showed an association between hypertension and advanced age (p < 0.001), prior diagnosis of hypertension (p < 0.001), use of ACEIs (p = 0.006), physical status (ASA) (p = 0.006), and size of the surgery (p = 0.028). Stratified analysis did not show an association between hypotension and use of ACE inhibitors. On logistic regression, the association between the size of the surgery and the age of the patient and the development of hypotension remained. CONCLUSIONS: We did not observe an association between the use of ACE inhibitors and hypotension during anesthesia. The authors considered that the recommendation to discontinue ACE inhibitors on the day before the surgery should be reviewed. Other studies are necessary to confirm those results. PMID- 20011861 TI - Main concerns of patients regarding the most common complications in the post anesthetic care unit. AB - BACKGROUND AND OBJECTIVES: 'Minor' events have a fundamental role in the determination of the quality of the Anesthesiology service. The objective of the present study was to evaluate the main concerns of patients regarding the post anesthetic period and to test the hypothesis that the most undesirable effects can be influenced by demographic characteristics. METHODS: Four hundred and forty patients answered a questionnaire immediately before the pre-anesthetic evaluation. Possible undesirable effects in the immediate postoperative period, based on data gathered in the literature considering the criterion of frequency but not severity were listed. Demographic data and the nine more frequent occupations were evaluated. The information gathered from the analysis of the questionnaires was related with anthropometric, socioeconomic, and educational data of the patients to assess their influence on the answer profile. RESULTS: Among the undesirable effects, the fear of being intubated upon waking up was mentioned more often as the most important factor, followed by 'severe pain at the surgery site'. Analysis of the three most frequent undesirable effects according to the demographic data did not show significant statistical differences, except for 'pain at the surgery site' (which was less common among male patients). CONCLUSIONS: The main post-anesthetic concerns of patients included: waking up with a tube in the throat, severe pain at the surgery site, and remembering being conscious during the surgery. Age, schooling, and family income did not determine differences in the concerns of patients. PMID- 20011862 TI - Influence of local anesthetics on the neuromuscular blockade produced by rocuronium: effects of lidocaine and 50% enantiomeric excess bupivacaine on the neuromuscular junction. AB - BACKGROUND AND OBJECTIVES: The effects of local anesthetics (LA) on neuromuscular transmission and their influence on the neuromuscular blockade produced by competitive neuromuscular blockers have not been fully investigated. The objective of this study was to evaluate, in vitro, the effects of lidocaine and 50% enantiomeric excess bupivacaine (S75-R25) on the neuromuscular blockade produced by rocuronium. METHODS: The rats were divided in five groups (n = 5) according to the drug used: isolated lidocaine, bupivacaine (S75-R25), or rocuronium (groups I, II, and II); and rocuronium in preparations previously exposed to LAs (groups IV and V). The concentrations used were as follows: 20 microg x mL(-1), 5 microg x mL(-1), and 4 microg x mL(-1) of lidocaine, bupivacaine (S75-R25), and rocuronium, respectively. The following parameters were evaluated: 1) the strength of muscular contraction of the diaphragm to indirect electrical stimulations, before and 60 minutes after the isolated addition of the LAs and rocuronium, and the association AL-rocuronium; and 2) the effects of LAs on membrane potential (MP) and miniature end-plate potentials (MEPP). The effect of LAs on muscle contraction in response to acetylcholine was evaluated in chick biventer cervicis preparations. RESULTS: Isolated lidocaine and bupivacaine (S75-R25) did not change the muscular response and the levels of MPs. In preparations exposed to LAs, rocuroniuminduced blockade was significantly greater than that produced by rocuronium alone. In chick biventer cervicis preparations, lidocaine and bupivacaine (S75R25) decreased contraction in response to acetylcholine. Lidocaine increased the frequency of MEPPs, which was followed by the blockade; bupivacaine (S75R25) caused a reduction in MEPPs followed by blockade. CONCLUSIONS: Local anesthetics caused a potentiation of the neuromuscular blockade produced by rocuronium. The results showed pre- and post synaptic effects. PMID- 20011863 TI - Total intravenous anesthesia with target-controlled infusion of remifentanil and propofol for ablation of atrial fibrillation. AB - BACKGROUND AND OBJECTIVES: Although ablation of atrial fibrillation (AF) is common in other centers, among us it is a new procedure. The choice of anesthesia, monitors, and anesthesiologic care for this procedure performed outside the surgical center has not been described. The objective of this report was to describe an anesthesia technique for ablation of AF. CASE REPORT: This is a 49-year old female weighing 73 kg, 155 cm, and ASA II due to hypertension. The patient was monitored with a 12-lead ECG, pulse oximetry, heart rate, bispectral electroencephalography for BIS measurement, suppression rate (SR), and SEF95, and mean arterial pressure (MAP). Intravenous target-controlled infusion (TCI) of propofol with a target of 4 microg x mL(-1), intravenous TCI of remifentanil with a target of 3 ng x mL(-1), and intravenous bolus of rocuronium 0.2 mg x kg(-1) were used for induction of anesthesia. The pharmacokinetic model of propofol described by Marsh was used and incorporated into the propofol PFS pump. The pharmacokinetic model of remifentanil described by Minto was incorporated into the Alaris PK infusion pump. Local effector, or biophase, concentrations corresponded to the information obtained from the infusion pumps and represented predictive measurements of the concentrations of both drugs on their sites of action. The concentrations of propofol and remifentanil were regulated according to BIS and MAP, respectively. CONCLUSIONS: Total intravenous anesthesia for ablation of AF can be a safe option considering the lack of electrophysiological changes in accessory pathways. The literature on this subject is scarce and new publications could justify, or not, this type of anesthesia during ablation of AF. PMID- 20011864 TI - Combined spinal epidural anesthesia during colon surgery in a high-risk patient: case report. AB - BACKGROUND AND OBJECTIVES: Combined spinal epidural anesthesia (CSEA) has advantages over single injection epidural or subarachnoid blockades. The objective of this report was to present a case in which segmental subarachnoid block can be an effective technique for gastrointestinal surgery with spontaneous respiration. CASE REPORT: Patient with physical status ASA III, with diabetes mellitus type II, hypertension, and chronic obstructive pulmonary disease was scheduled for resection of a right colon tumor. Combined spinal epidural block was performed in the T5-T6 space and 8 mg of 0.5% isobaric bupivacaine with 50 microg of morphine were injected in the subarachnoid space. The epidural catheter (20G) was introduced four centimeters in the cephalad direction. Sedation was achieved with fractionated doses of 1 mg of midazolam (total of 6 mg). A bolus of 25 mg of 0.5% bupivacaine was administered through the catheter two hours after the subarachnoid block. Vasopressors and atropine were not used. CONCLUSIONS: This case provides evidence that segmental spinal block can be the anesthetic technique used in gastrointestinal surgeries with spontaneous respiration. PMID- 20011865 TI - Anesthetic conduct in cesarean section in a parturient with unruptured intracranial aneurysm. AB - BACKGROUND AND OBJECTIVES: The anesthetic management of a parturient with unruptured intracranial aneurysm scheduled to undergo cesarean section is interesting, since it has several particularities associated with pregnancy related physiologic changes that are associated with the risk of aneurismal rupture during the anesthetic procedure. Studies on this subject are rare in the literature and, therefore, the dissemination of those cases is important. CASE REPORT: This is a 31-year old female at term with unruptured intracranial aneurysm scheduled for cesarean section under epidural block. The procedure evolved without maternal or fetal intercurrences. CONCLUSIONS: Evidence-based recommendations for obstetric anesthesia in patients with unruptured intracranial aneurysm are lacking. Experimental or clinical data confirming or refuting general anesthesia or regional blocks in this context do not exist. Thus, the decision of which technique should be used is individual, considering the risks and benefits of each procedure and the experience of the anesthesiologist. PMID- 20011866 TI - Transesophageal echocardiography and intraoperative diagnosis of persistent left superior vena cava. AB - BACKGROUND AND OBJECTIVES: Isolated persistent left superior vena cava has an incidence of 0.5% in the normal population, but in patients with congenital cardiopathy its incidence ranges from 3% to 10%. The objective of this report was to present a case of intraoperative diagnosis with transesophageal echocardiography and to emphasize the importance of its routine use in surgical procedures for correction of congenital cardiopathies. CASE REPORT: This is a 16 year old male patient, ASA II, with a diagnosis of superior sinus venosus interatrial communication (IAC) with partial anomalous drainage of the pulmonary veins scheduled for the surgical correction of the cardiopathy. After induction of general anesthesia, transesophageal echocardiography (TEE) showed initially a dilation of the right cardiac chambers, a 17-mm superior sinus venosus IAC, anomalous drainage of the right superior pulmonary vein draining into the right superior vena cava (SVC), and dilation of the coronary sinus raising the possibility of persistent left superior vena cava. To confirm this suspicion, 10 mL of NS (works as a contrast in echocardiography) were injected in the venous access of the left arm, and microbubbles crossing the coronary sinus were observed immediately afterwards confirming the diagnosis of persistent left superior vena cava. CONCLUSIONS: Routine intraoperative transesophageal echocardiography in patients with congenital cardiopathy is a fundamental auxiliary method, not only for the surgeon, often having direct influence on the surgical technique used, but also for the anesthesiologist, who can get useful information for the hemodynamic management of the patient. PMID- 20011867 TI - Ways the anesthesiologist can contribute to the prophylaxis of infection in the surgical patient. AB - BACKGROUND AND OBJECTIVES: Besides the increase in hospital costs, surgical patients with nosocomial infection may not recover their previous condition and their condition can even evolve to death. Besides well-established patient related risks, postoperative infection is also dependent on the surgical procedure, the hospital conditions and the surgeon. Despite several protocols to prevent nosocomial infections, standardization of intraoperative interventions to optimize those patients does not exist. Thus, the objective of this review was to discuss some of those interventions that are effective and necessary, alerting anesthesiologists of their importance in preventing nosocomial infections. CONTENTS: In this review, the causes of infections in surgical patients as well as the adequate administration of antibiotics, temperature and blood glucose control, and strategies of mechanical ventilation, hydration, and transfusions that can reduce the incidence of infections in surgical patients are discussed. CONCLUSIONS: The anesthesiologist is the professional who should intervene in the intraoperative period with simple measures to optimize the care of surgical patients and to reduce the incidence of infections. PMID- 20011868 TI - The pediatric cardiac surgery as a philanthropic activity in the country and humanitarian mission abroad. PMID- 20011869 TI - The new Qualis, which has nothing to do with the science of Brazil: open letter to the president of CAPES. PMID- 20011871 TI - Journal impact factor: this editorial, academic and scientific influence. AB - In this report the authors present information on the bibliometric instruments and their importance in measuring the quality of scientific journals and researchers. They in particular the history and deployment of the impact factor of the existing Institute for Scientific Information since 1955. Are presented and discussed the criticism regarding the inadequacy of the impact factor for evaluation of scientific production, misuse and strategies editorial handling of the bibliometric index. It is presented to the new classification CAPES for the journals, based on various criteria and the impact factor and its influence on national scientific and academic life. The authors conclude that, despite all obstacles and discussions, the impact factor of the Institute for Scientific Information is still an useful tool and the only isolation available to assess the scientific and intellectual productivity. PMID- 20011872 TI - Methylene blue for vasoplegic syndrome treatment in heart surgery: fifteen years of questions, answers, doubts and certainties. AB - OBJECTIVE: There is strong evidence that methylene blue (MB), an inhibitor of guanylate cyclase, is an excellent therapeutic option for vasoplegic syndrome (VS) treatment in heart surgery. The aim of this article is to review the MB's therapeutic function in the vasoplegic syndrome treatment. METHODS: Fifteen years of literature review. RESULTS: 1) Heparin and ACE inhibitors are risk factors; 2) In the recommended doses it is safe (the lethal dose is 40 mg/kg); 3) The use of MB does not cause endothelial dysfunction; 4) The MB effect appears in cases of nitric oxide (NO) up-regulation; 5) MB is not a vasoconstrictor, by blocking of the GMPc system it releases the AMPc system, facilitating the norepinephrine vasoconstrictor effect; 6) The most used dosage is 2 mg/kg as IV bolus followed by the same continuous infusion because plasmatic concentrations strongly decays in the first 40 minutes; 7) There is a possible 'window of opportunity' for the MB's effectiveness. CONCLUSIONS: Although there are no definitive multicentric studies, the MB used to treat heart surgery VS, at the present time, is the best, safest and cheapest option, being a Brazilian contribution for the heart surgery. PMID- 20011873 TI - Common usage of cardiologic anatomical terminology: critical analysis and a trilingual discussion proposal. AB - BACKGROUND: Terminology and Lexicography have been especially addressed to the Allied Health Sciences regarding discussion of case reports or concerning publication of scientific articles. The knowledge of Human Anatomy enables the understanding of medical terms and the refinement of Medical Terminology makes possible a better anatomicomedical communication in a highly technical level. Most of the scientific publications in both Anatomy and Medicine are found only in English and most of dictionaries or search resources available do not have specificity enough to explain anatomicomedical, terminological, or lexicographical occurrences. OBJECTIVE: To design and produce a multilingual terminological dictionary (Latin-English-Portuguese-Spanish) containing a list of English anatomicomedical terms in common usage in cardiology subspecialties addressed to medical students and professionals, to other allied health sciences professionals, and to translators working in this specific field. METHODS: Terms, semantical and grammatical components were selected to compose an anatomicocardiological corpus. The adequacy to the thematic terminological research requests and the translation reliability level will be settled from the terminology specificity in contrast to the semantics, as well as from a peer survey of the main terms used by national and international experts in specialized journals, Internet sites, and from text-books on Anatomy and Cardiology. The inclusion criteria will be the terms included in the English, Portuguese, and Spanish Terminologia Anatomica - the official terminology of the anatomical sciences; nonofficial technical commonly used terms which lead to terminology or translation misunderstanding often being a source of confusion. RESULTS: A table with a sample of the 508 most used anatomical cardiologic terms in English language peer-reviewed journals of cardiology and (pediatric and adult) thoracic surgery is shown. CONCLUSIONS: The working up of a multilingual terminological dictionary reduces the risk of ambiguities, inconsistencies, inutilities, and repetitions concerning the Nomenclature addressed to the Allied Health Sciences by prioritizing the inclusion of official technical terms and a judicious selection of commonly used terms. Efforts to standardize lists of structures in Humana Anatomy lead to both opportunities of scientific update and conceptual enlightenment. PMID- 20011874 TI - Prevalence and risk factors for acute renal failure in the postoperative of coronary artery bypass grafting. AB - OBJECTIVE: To determine the prevalence, risk factors, and the clinical outcome of patients undergone coronary artery bypass grafting who progressed with Acute Renal Failure (ARF). METHODS: A retrospective cohort prospective study was performed from data of 186 patients undergone surgery from January 2003 through June 2006. The stored data were analyzed using the software STATA 9.0. RESULTS: The prevalence of ARF was of 30.6% (57/186). In 7.0% (4/57) dialysis therapy was needed. The mean age of patients with and without ARF progression was 62.8 (+/ 9.4) years and 61.3 (+/-8.8) years respectively (P=NS). CPB time >115 min (p= 0.011) and cross-clamp time >85 min (p=0.044) were related to ARF by the univariate analysis. The need for intra-aortic balloon (P= 0.049), mechanical ventilation >24h (P = 0.006), Intensive Care Unit (ICU) stay > three days (P< 0.0001), bradycardia (P= 0.002), hypotension (P= 0.045), arrhythmia (P=0.005) and inotropic infusion (P= 0.0001) were higher in the ARF group. Only the ICU stay longer > 3 days showed statistical correlation with ARF by the multivariate analysis (P=0.018). The mortality rate with and without ARF was 8.8% (five cases) and 0.8% (one case) respectively (P=0.016), but it reached 50% (2/4) in dialytic patients. CONCLUSION: ARF was a frequent and severe postoperative complication associated with higher mortality and longer ICU stay, which presented as risk factors: longer CPB and cross-clamp times, mechanical ventilation > 24h and hemodynamic instability. PMID- 20011875 TI - Cystatin C and glomerular filtration rate in the cardiac surgery with cardiopulmonary bypass. AB - OBJECTIVE: The aim of this study was to compare cystatin C versus creatinine as a marker for acute kidney injury in patients submitted to cardiac surgery with cardiopulmonary bypass. METHODS: Fifty consecutive patients submitted to coronary artery bypass grafting were studied. Renal function was evaluated by serum cystatin C and creatinine. Blood samples were obtained from each patient at three time points: before operation, and on the first and fifth postoperative days. Glomerular filtration rate (GFR) was calculated by Cockcroft-Gault (CG), Modification of Diet in Renal Disease (MDRD), and Larsson (Cys-GFR) formulas. RESULTS: Creatinine and GFR by CG and MDRD formulas did not show statistical difference between study times. After renal injury from surgery, there was an increase in cystatin C on the 1st and 5th day after surgery, being significantly different on the 5th postoperative (P<0.01). The GFR by Larson formula was higher in the preoperative time (105.2 +/- 41.0 ml/min) than in the 5th postoperative day (89.5+/- 31.5 ml/min; P<0.012). CONCLUSION: The cystatin C and the Cys-GFR showed significant changes after cardiac surgery when compared with the creatinine and respective GFR calculated by the Cockcroft-Gault and MDRD formulas. PMID- 20011876 TI - Expression of apoptosis in human saphenous vein grafts in restoration of blood flow through coronary bypass surgery. AB - OBJECTIVE: To investigate the possible role of apoptosis on brief distensions of human saphenous veins at different pressures. METHODS: Fresh isolated grafts of human saphenous vein were assigned as control or distended (D) for fifteen seconds at 100, 200 and 300 mmHg. The degree of apoptotic caspases 3, 8, 9 and anti-apoptotic protein Bcl-2 expression were assessed by immunohistochemistry. RESULTS: Fresh isolated segments of distended human saphenous veins presented similar apoptotic protein expression when compared with control veins. However, the Bcl-2 expression was significantly higher in the 300 mmHg distended segments compared with the control vein. CONCLUSION: These findings show that intact segments of human saphenous veins submitted to distensions at different pressures have similar apoptotic proteins expression when compared with non-distended control veins. Therefore, brief distensions commonly performed during surgical harvesting do not trigger apoptosis, and probably are not involved on the physiopathological mechanisms that lead to graft failure. PMID- 20011877 TI - Video-assisted cardiac surgery: results from a pioneer project in Brazil. AB - OBJECTIVE: To demonstrate the possibilities of the use of videothoracoscopy in cardiac surgery using cardiopulmonary bypass (CPB). METHODS: Between February 2006 and November 2008, 102 patients underwent consecutively minimally invasive video-assisted cardiac surgery. The cardiac pathologies approached were: mitral valvopathy (n=56), aortic (n=14), interatrial communication (IC) (n=32), six patients presented associated tricuspid insufficiency and 12 presented atrial fibrillation. The age ranged from 18 to 68 years and 57 were female. The surgical approach was: femoral arterial and venous cannulation, minithoracotomy ranging from four to six centimeters (cm) at the level of the 3 degrees or 4 degrees right intercostal space (RICS), depending on the pathology of the patient, between anterior axillary line and hemiclavicular line, submammary or right periareolar groove through the right breast and thoracoscopy. RESULTS: The surgical procedures were: plasty (n=20) or mitral valve replacement (n=36), aortic valve replacement (n=14), atrioseptoplasty using pericardial patch (n=32), tricuspid valve repair with rigid ring (n=6) and surgical correction of atrial fibrillation with radiofrequency (n=12). There were no complications during the procedures. There was no conversion to thoracotomy in neither case. Two patients developed atrial fibrillation in the postoperative period. There was an episode of stroke seven days after the hospital discharge and one death (0.9%) due to systemic inflammatory response syndrome (SIRS). CONCLUSION: This study demonstrates the coverage of pathologies that are possible to be approached by video-assisted cardiac surgery with cardiopulmonary bypass being a safe and effective procedure with low morbimortality. Minimally invasive video-assisted cardiac surgery is already a reality in Brazil, demonstrating excellent aesthetic and functional results. PMID- 20011878 TI - Pulmonary artery banding: a simple procedure? A critical analysis at a tertiary center. AB - OBJECTIVE: Although pulmonary artery banding (PAB) seems to be a technically simple procedure it presents several peculiarities and is related to a significant morbidity and mortality. The aim of this study is to analyze the experience of a tertiary hospital on the PAB by assessing and correlating many aspects related to the procedure. METHODS: Between January 2000 and December 2008, 61 patients undergone PAB due to congenital heart disease with increased pulmonary blood flow at Messejana Heart Hospital were assessed as for mortality, complications, stay in mechanical ventilation and need for intensive care unit (ICU), use of vasoactive drugs, difficulties in the adjustment on the banding and reoperations. Some statistical analyzes were performed to compare the subgroups. RESULTS: In 46.8% of the patients the intended pressoric adjustment was not achieved and in 6.5% it was necessary another surgery to readjust the banding. The mean time of mechanical ventilation was 14.1+/-49.6 days and ICU 14.16+/ 10.92 days. In 82.6% of the patients vasoactives drugs were administrated for 10.3+/-12.79 days. Severe complications were noted in 49.15% of patients and cardiac insufficiency was the most common one with an incidence of 44%. The mortality rate was 8.2% and it was not influenced by weight or associated procedures with the PAB neither if univentricular or biventricular heart disease. CONCLUSION: The PAB can be performed with acceptable mortality rates compatible with the ones of the world literature. Nevertheless, the adjustment of the banding is difficult to be assessed during the surgery by making the procedure complex and justifying the high incidence of complications and long stay in ICU. It wasn't found any specific risk factor significant to mortality neither uni- or biventricular heart disease. PMID- 20011880 TI - Risk factors for hospital mortality in valve replacement with mechanical prosthesis. AB - OBJECTIVE: Identification of risk factors for cardiac surgery can improve surgical results. Our aim is to identify factors related to increased hospital mortality for patients who underwent mechanical cardiac prosthesis implant. METHODS: Prospective study with retrospective data acquirement study including 335 consecutive patients who underwent at least one implant of St. Jude Medical mechanical prosthesis between December 1994 and September 2005 at the Cardiology Institute of RS. Valve implants were 158 (47.1%) in aortic position, 146 (43.6%) in mitral and 31 (9.3%) in aortic and mitral. The following characteristics were analyzed in relation to hospital death: gender, age, body mass index, NYHA functional class, ejection fraction, type of valve lesion, hypertension, diabetes mellitus, serum creatinine, preoperative arrhythmias, prior heart surgery, CABG surgery, concomitant tricuspid valve surgery and operative priority (elective, urgent or emergent). Logistical regression was used to analyze data and odds ratio was calculated for individual factors. RESULTS: During the follow-up there were 13 (3.88%) deaths. In-hospital mortality risk was associated with serum creatinine (P<0.05), ejection fraction < 30% (P<0.001), mitral valve lesion (P<0.05), concomitant CABG surgery (P<0.01), prior cardiac surgery (P<0.01) and reoperation (P<0.01). Increased odd-ratio were related to previous cardiac surgery (5.36; IC95% 0.94-30.56), combined revascularization (5.28; IC95% 1.51 18.36), valvar reoperation (4.69; IC95% 0.93-23.57) and concomitant tricuspid annuloplasty (3.72; IC95% 0.75-18.30). CONCLUSION: The mortality rate is within the parameters found in the literature, identifying recognized factors which neutralization by changes in surgical indication and medical management may enable risk reduction. PMID- 20011881 TI - Topical use of antifibrinolytic agent to reduce postoperative bleeding after coronary artery bypass surgery. AB - OBJECTIVE: Antifibrinolytic agents reduce bleeding after cardiac surgery, but there are adverse effects after their systemic use. These effects are avoided by topical application of antifibrinolytic agents in pericardial cavity. We compared the effects of topically applied epsilon-aminocaproic acid (EACA) and placebo on postoperative bleeding and transfusion requirements after coronary artery bypass surgery. METHODS: In this single center prospective, randomized, double-blind trial, 53 patients were randomized into two groups to receive EACA (24 g in 250 ml of saline solution) or placebo (250 ml of saline solution) before sternal closure. Groups were comparable with respect to all preoperative and intraoperative variables. Postoperative bleeding, transfusion requirements and hematologic parameters were evaluated. RESULTS: Postoperative bleeding within first 24 hours (h) period (EACA group 154.66+/-74.64 x Placebo group 220.21+/ 136.42 ml; P=0.031) showed statistically significant inter-group difference, within 48 h (EACA group 259.14+/-420.07 x Placebo group 141.67+/-142.58 ml; P=0.614), as well as cumulative blood loss (EACA group 832.07+/-576.86 x Placebo group 827.50+/-434.12 ml; P=0.975), not showed statistically inter-group differences. Inter-group difference of blood product requirements was statistically significant (EACA group 185.90+/-342.07 x Placebo group 439.42+/ 349.07 ml; P=0.016). Laboratory analyses showed no differences between the two groups postoperative (hematologic characteristics: hemoglobin (g/dl)- EACA group 9.18+/-0.92 x Placebo group 8.85+/-1.48 g/dL; P=0.11; hematocrit (%)-EACA group 28.15+/-3.35 x Placebo group 26.67+/-4.15%; P=0.06). CONCLUSION: Topical use of epsilon aminocaproic acid reduces postoperative bleeding in the first 24 hours and requirements of blood transfusion after coronary artery bypass graft surgery. PMID- 20011882 TI - Outcomes of aortic coarctation surgical treatment in adults. AB - OBJECTIVE: The aim of this study is to describe our experience in aortic coarctation surgery in adult patients by assessing the immediate and mid-term outcomes. METHODS: From January 1987 to March 2000, 50 consecutive adult patients underwent surgery for correction of aortic coarctation, through left lateral thoracotomy. Of these, forty two (84%) patients presented high blood pressure, with mean systolic arterial pressure of 170.56 mmHg (125-220 mmHg). The mean of pressure gradient in the coarctation area was 51.4 mmHg (18-123 mmHg). Other associated surgical cardiovascular diseases were not treated in the same operative act, except in two cases of patent ductus arteriosus (PDA). Three different techniques were used: aortic coarctation resection with end-to-end anastomosis was performed in 20 (40%) patients, coarctation enlargement with bovine pericardial patch was performed in 22 (44%) patients and synthetic tube interposition was performed in eight (16%) patients. RESULTS: Operative morbidity was low; there was one case of bleeding who required reoperation. The most common immediate postoperative event was high blood pressure (98%), but it was easily controlled by intravenous drugs. There was no hospital death. Mean residual pressure gradient was 18.7 (8-33 mmHg). Patients were discharged in 9.5 days (5 30). Postoperative follow-up mean was 46.8 months (1-145 months) in 45 (91.8%) patients. Forty one (91.1%) of these followed-up patients had normal blood pressure, whereas 75.6% of them without drugs intake. 93.3% of these followed-up patients were asymptomatic. Four of them required further surgical operation, one needed a pacemaker implant, other two patients needed a cardiac valve replacement and one had endocarditis. There was one related death due to sepsis secondary to endocarditis. CONCLUSION: Surgical treatment of aortic coarctation, even in adult patients, is an efficient therapeutic choice, regardless of the applied surgical technique, with low morbidity and mortality. It reduces efficiently the arterial pressure levels in both immediate and mid-term follow-up. PMID- 20011883 TI - Intermittent annular reduction with Alfieri's repair in the treatment of mitral insufficiency in children: initial results. AB - OBJECTIVE: Mitral valve insufficiency must be treated by repair whenever as possible as it provides better results - especially within the pediatric population in order to avoid valve replacement inconvenients. The aim of this study is to describe mitral valve repair technique based on an anatomical and functional approach. METHODS: During a 13 months period, eight children (age ranged from 2 and 12 years old 6.37 +/- 3.66) with mitral valve insufficiency underwent intermittent anterior and posterior annuloplasty associated with Alfieri's repair - performed by placing a stitch between the anterior and posterior leaflets in the point of maximal regurgitation thus creating a double orificial mitral valve. RESULTS: There were no surgical deaths. All patients underwent postoperative echocardiography. Three patients showed mild mitral valve insufficiency and five patients showed no insufficiency. The time in intensive care unit ranged from 2 to 4 days (2.5 +/- 0.75), and the time of stay in hospital ranged from 5 to 8 days (6.37 +/- 1.06). CONCLUSION: In spite of the little sample size, the proposed technique proved itself to be simple and effective in the treatment of single mitral valve insufficiency within the pediatric population. In addition, it also yielded satisfactory immediate results. Long-term follow-up is nevertheless necessary in order to evaluate long term results. PMID- 20011884 TI - Effects of physiotherapeutic instructions on anxiety of CABG patients. AB - INTRODUCTION: The coronary artery bypass graft surgery is an option of treatment for cardiovascular diseases, and the patients who underwent such procedure can suffer from preoperative anxiety. OBJECTIVE: The aim of this study is to verify the effects of physiotherapeutic instructions at the anxiety level on patients undergone coronary artery bypass graft surgery. METHODS: Randomized clinical trial, with the sample composed by 51 individuals, 27 on the control group and 24 on the intervention; on which it was assessed the anxiety (Beck Scale for Anxiety) and pain (Visual Analog Scale) levels on the pre- and postoperative period and only the second group received instructions about the surgery procedures and ventilatory exercises. For statistical analysis the Wilcoxon, Mann Whitney and Spearman tests were used. RESULTS: The lower anxiety levels were observed in patients that received the intervention on the period before surgery (9.6 +/- 7.2 versus 13.4 +/- 5.9, P=0.02). On the control group, the difference among the anxiety levels before and after the surgery was statistically significant (P= 0.003). The female individuals presented more anxious before the surgery compared to the male ones (P=0.058). It was also verified a relationship between length of stay at hospital and postoperative anxiety (P=0.05), where the more anxious individuals remained more time at the hospital. CONCLUSION: Patients oriented and instructed about physiotherapeutic ventilatory exercises and hospital routine, presented their levels of anxiety reduced on the preoperative compared to the control group. However, at the postoperative, both groups presented their anxiety levels reduced without significant difference between them. PMID- 20011885 TI - Vehicle influence on potassium replacement effectiveness in hypokalemic rats. AB - INTRODUCTION: Patients who undergo cardiac surgery are commonly treated with diuretic therapy for the management of volume overload. The concern of hypokalemia important in the adult population submitted to cardiac surgery has been described. Intravenous potassium (K+) replacement dilution is only recommended with sodium chloride 0.9% solution (SF0.9%), likely due to the putative effects of glucose solution 5% (SG5%) on insulin secretion, which influence K+ replacement quality. However, it is not yet experimentally proved the influence of SF0.9% and SG5% on K+ replacement quality. OBJECTIVES: To evaluate the effects of different vehicles of K+ replacement on blood K+ levels in furosemide hypokalemic rats. METHODS: Male Wistar rats divided into four groups: K++SF, K++SG, SF and SG. Jugular vein was cannulation for K+ replacement and femoral vein was cannulated for blood analysis were performed. Furosemide (50mg/kg) was injected S.C. to induce hypokalemia. It was analyzed potassium plasmatic levels 24 hours before furosemide injection, 24 hours after furosemide injection and 30 minutes after post-replacement. RESULTS: There was no significative difference in blood K+ levels when compared to the basal values (pre-furosemide) in all groups. However, the levels [K+] returned to baseline in both groups receiving K++SF or K++SG, which was not observed in groups receiving only SF and SG. Only K+SF presented increased after K+ replacement (P<0.05). CONCLUSION: K+ replacement diluted both in SF and SG did not affect blood K+ levels in rats. PMID- 20011886 TI - Does aprotinin preserve platelets in children with acyanogenic congenital heart disease undergone surgery with cardiopulmonary bypass? AB - OBJECTIVE: Evaluation of the hemostatic and platelets effects in children with acyanogenic congenital heart disease undergone on-pump surgery who received aprotinin. METHODS: A prospective randomized study was performed on children aged 30 days to 4 years who had undergone correction of acyanogenic congenital heart disease using cardiopulmonary bypass (CPB) and were divided into two groups: Control (n=9) and Aprotinin (n=10). In the Aprotinin Group the drug was administered before and during CPB and the hemostatic dysfunction was analyzed by clinical and biochemical markers. Differences were considered to be significant when P<0.05. RESULTS: The groups were similar regarding demographic and intraoperative variables, except for a greater hemodilution in the Aprotinin Group. The drug presented no benefit regarding time of mechanical pulmonary ventilation, stay in the postoperative intensive care unit and hospital, or regarding the use of inotropic drugs and renal function. Platelet concentration was preserved with the use of Aprotinin, whereas thrombocytopenia occurred in the Control Group since the initiation of CPB. Blood loss was similar for both groups. There were no complications with the use of Aprotinin. CONCLUSION: Aprotinin quantitatively preserved the blood platelets in children with acyanogenic congenital heart disease. PMID- 20011887 TI - Identification of a statistical method as a quality tool: patient's length of stay in the operating room. AB - OBJECTIVE: To identify a statistical method that may express the patient length of stay in the operating room and build a 'matrix of relationship' for optimizing this time, the real and exact time of the operation. METHODS: The analysis of survival and the Kaplan-Meier estimator allowed to calculate the survival curves for different times and the 'matrix of relationship' with 10 hypothesis to help in choosing the new operation. The study consisted of a simple random sample of 71 patients, from elective operations for adults in Cardiac Surgery/Clinics Hospital/Unicamp, with confidence level of 95% in 2008. RESULTS: On average, the times of the operations over at a range of 140 minutes to 200 minutes and excess from 5 minutes to 90 minutes. In general, on average, one operation was daily performed within 520 minutes, for a time of 720 minutes. CONCLUSION: 1) With the maximum available time of 720 minutes is not possible to perform surgery, unless using the 'matrix of relationship', whereas the maximum time available varies between 660 minutes and 690 minutes, considering the range of cleaning of the room. 2) The time of the patient in the operating room is a time that includes the time of learning by the student in an university hospital school. 3) When optimizing the time, most patients will benefit, causing a decrease from the waiting list for new opeartions. 4) The 'matrix of relationship' allows to view and express opinion on a better decision making in addition to decide upon several assumptions. PMID- 20011888 TI - The role of electroanalgesia in patients undergoing coronary artery bypass surgery. AB - OBJECTIVE: To assess the electroanalgesia as an effective method in the reduction of pain and consequent improvement in lung function in patients undergoing coronary artery bypass graft surgery. METHODS: During the period of one year were studied 30 patients undergoing surgery for treatment of the ischemic coronary disease. After randomization, 15 patients were allocated in the study group (that received electroanalgesia) and 15 patients from the controlled group (placebo). From the 1st until the 5th postoperative day were realized two applications of electroanalgesia or the placebo current, according the group where the patient was allocate. RESULTS: There was a reduction in the levels of pain in the study group compared to the controlled group, evidenced by the value P<0.05; however, there isn't evidence of statistical difference of the spirometric variables between them. CONCLUSION: The group that did electroanalgesia presented reduction in the intensity of postoperative pain, which however did not mean improvement in respiratory function of these patients. PMID- 20011889 TI - Importance of pre- and postoperative physiotherapy in pediatric cardiac surgery. AB - Lung complications during postoperative of pediatric heart surgery are frequently highlighting atelectasis and pneumonia. Physiotherapy has an important role in the treatment of these complications. We reviewed and update the physiotherapy performance in the preoperative and in the postoperative lung complication of pediatric cardiac surgery. We noted efficacy of physiotherapy treatment through different specific techniques and the need for development of new studies. PMID- 20011890 TI - Right coronary bypass grafting with coronary-coronary venous bypass graft. AB - OBJECTIVE: The following report describes the experience of the Cardiovascular Surgery Team of the ABC Medical School with coronary-coronary bypass grafting. METHODS: Four patients undergone off-pump coronary artery bypass grafting, with coronary-coronary bypass of the right coronary artery, by using great saphenous vein graft. RESULTS: The follow-up period was 3 to 5 years without any intra- or postoperative recurrence. One patient presented anginal symptoms four years after the surgery and underwent cinecoronariography that revealed patent anastomosis. CONCLUSION: The coronary-coronary bypass is a good option for patients undergone coronary artery bypass grafting. PMID- 20011891 TI - Axillary artery in cardiopulmonary bypass: indications and results. AB - OBJECTIVES: To determine indications and results of axillary artery cannulation for cardiopulmonary bypass. METHODS: From January 2005 through December 2008, axillary artery cannulation was used in 48 patients. Mean age was 62 +/- 11 years and 33 (69%) patients were males. Axillary artery was approached by infraclavicular incision and the cannula introduced in a 8 millimeter Dacron side graft. RESULTS: Indications were calcified aorta (N=18, 38%), aortic dissection (N=15, 31%), ascending and/or aortic arch aneurysm (N=11, 23%) and prior to reoperative median sternotomy (N=4, 8%). Changes in intraoperative planning occurred most often in patients with calcified aorta (100% versus 10%, P<0.0001) than in patients with other indications, which follow their preoperative plan. Cardiopulmonary bypass (deep hypothermic circulatory arrest in 55% and conventional in the remaining) was uneventfully conducted in all patients but one (success rate 98%) due to undiagnosed inominate artery stenosis. Local complication was lymphatic drainage in three (6.2%) patients. CONCLUSIONS: Axillary artery is an alternative cannulation site in patients unsuitable to aortic cannulation. The type of indication may determine intraoperative changes in surgical planning. PMID- 20011892 TI - Heart transplantation in primary amyloidosis. AB - Cardiac amyloidosis is a disease with a gloom life expectancy after the beginning of the symptomatic phase, usually with sudden death as the final event. The aggression to other organs, although, can make heart transplantation a disputable form of treatment taking into consideration the shortage of donor organs. The aim is to report the evolution with a survival of seven years after heart transplantation and in very fair condition of a patient with amyloidosis. One year after the heart transplantation, there was indication of renal transplantation also from the aggression from the disease. This patient compares' favorable with three other patients also from our service, who died early after de diagnosis. Even considering the multi systemic nature of amyloidosis, we can accept that in peculiar patients justified the heart transplantation, taking in the consideration the very bad prognosis of the disease. PMID- 20011893 TI - Giant coronary artery dilatation in an asymptomatic patient. AB - A case of giant dilatation of coronary arteries is presented, with review of the literature on the subject, and discussions about management of the patients with such disease. PMID- 20011894 TI - Successful medical management of prosthetic-valve endocarditis complicated by perivalvular abscess. AB - We present a case of a 44-year-old man with prosthetic aortic endocarditis complicated by a perivalvular abscess. He evolved with improvement of the infectious process only under clinical treatment. The patient presented a prior history of rheumatic fever and had previously been undergone three valve replacements due to prosthesis dysfunction and previous endocarditis. In this case report we discuss the main features of perivalvular abscess complicating infective endocarditis. PMID- 20011895 TI - Total correction of tetralogy of Fallot in child with left pulmonary artery agenesis. PMID- 20011896 TI - Atrioventricular septal defect with tetralogy of Fallot in patient with Down's syndrome. PMID- 20011897 TI - Video-assisted minimally invasive mitral valve repair: periareolar approach. PMID- 20011898 TI - Research on the drugs phenomenon from the perspective of multicenter studies in Latin America and the Caribbean. PMID- 20011899 TI - Illicit drug use in seven Latin American countries: critical perspectives of families and familiars. AB - This cross-sectional multi-centre study explored how family members and friends of illicit drug users perceived protective and risk factors, treatment facilities and policies and laws regarding illicit drug use. Family members and friends of illicit drug users were recruited in 10 urban health care outpatient units in 7 Latin American countries (Brazil, Colombia, Costa Rica, Ecuador, Guatemala, Honduras and Mexico) to complete a questionnaire. The majority of the respondents chose psycho-social factors over genetic or biological explanations as causes of drug problems. Respondents felt that families and governments were responsible for preventing drug problems. Church/religious institutions were most often mentioned in the context of accessible treatment. When asked about access to treatment facilities, the majority said that there were not enough. Shame about drug use, cost, and limited treatment options were most often cited as barriers to treatment. PMID- 20011900 TI - [Critical perspective of the family and acquaintances on family and community risk factors in illicit drug use in Sao Jose, Costa Rica]. AB - This article presents the results of a quantitative study. The purpose was to describe the perspective of family members and acquaintances of illicit drug users about family and community risk factors that can contribute to addiction to illicit drugs. A questionnaire was used for data collection. The population consisted of 100 individuals, 18 years of age or older, who reported being affected by their relationship with an illicit drug user (relative or acquaintance). Most users (82%) were men, with an average age of 27.3 years. Family risk factors included: family rejection (99%), not feeling loved (98%), lack of communication (95%), family conflicts and violence (95%). Social or community factors included: 99% having friends who use drugs, 99% peer pressure, 93% living in an unsafe area, and 99% experienced a stressful event. The critical perspective proposes to use more prevention strategies to avoid risk factors in the family and community. PMID- 20011901 TI - [Use of illicit drugs and critical perspectives of drug users' relatives and acquaintances in Northern Rio de Janeiro City, Brazil]. AB - This article presents the partial results of a multicenter, qualitative study, which involved seven Latin-American countries and Canada. The results presented refer to Northern Rio de Janeiro (city), Brazil. The objective of the study was to describe the perspective of relatives/acquaintances of illicit drug users about protective and risk factors, prevention initiatives, treatment services, and legal aspects regarding illicit drugs. Interviews were performed with 99 individuals, who reported being affected by their relationship with an illicit drug user (relative or acquaintance), approaching their perspectives regarding the key-domains. Most participants were women (73.7%); relatives who used drugs were mostly men (78.2%); the most consumed drug was marijuana (77.8%). The highlighted protective factor was having recreational-sports activities in the community (88.9%), and the risk factor was curiosity for trying something new (94.4%). The main treatment services were Church Groups (51.5%), and participants stated that laws should be more punitive (82.8%). In conclusion, this information is essential to fight against drug use/abuse, showing that there is a need for actions that consider different perspectives at different levels. PMID- 20011902 TI - [Illegal aspects of illicit drugs use in Mexico]. AB - The objective of this cross-sectional, descriptive study was to obtain the opinions of a group of people about legal issues regarding addiction. Data collection was performed using a structured questionnaire with four themes. In order to participate, the individual could not use any drugs but should have a close relationship with a drug user. The data was processed using SPSS V. 14. There were 100 participants, 75% of whom were women, and 38% had a drug user as a friend, mainly cocaine and marijuana users. The participants had one opinion in common: laws should be more severe for people who use, sell, or transport drugs. The current laws do not improve consumers' criminal behavior. There is a need for further studies addressing people's opinion about this phenomenon in order to obtain a more realistic view of this drug issue. PMID- 20011903 TI - [Drug use and treatment from the perspective of the users' family and friends in Bogota, Colombia]. AB - This article presents an analysis of the partial results from a quantitative study that addresses the perspective of drug users' family and friends regarding the treatment of problems resulting from the use of illicit drugs. Participants were 104 health service users in Bogota. Of the participants, 58% consider that drug use is a disease that needs treatment; 56% stated the general hospital as the main alternative for treatment; 95% reported that the state accounted for this situation. Participants were more acquainted with private institutions, with therapeutic communities and religious groups being reported as the ones with the strongest commitment to the problem. For 73% of participants, the existing services are inappropriate and of difficult use and access. Stigma is the main barrier for a person to seek health care, and only 50% believe there is any use in treatments. There is a need for broader dissemination of the existing resources so as to increase their implementation and use. PMID- 20011904 TI - [Risk factors related to the use of illegal drugs: the critical perspective of drug users' relatives and acquaintances at a public health center in San Pedro Sula, Honduras]. AB - This article presents quantitative data from a multicenter, cross-sectional study, which was performed at a public health center in San Pedro Sula, Honduras, using multiple methods. The objective of the study was to describe the critical perspective of people who reported being affected by their relationship with an illicit drug user (relative or acquaintance) in terms of risk factors. Data collection was performed using 100 questionnaires. Most participants were women with low education levels. Drug users were mostly men, with an average age of 23.3 years. The most consumed drug was marijuana (78%), followed by crack/cocaine (72%), glue/inhalants (27%), hallucinogens (ecstasy/LSD) (3%), amphetamines/stimulants (1%), and heroin (1%). The identified risk factors include: previous experience with alcohol/tobacco, having friends who use drugs, lack of information, low self-esteem, age, and other personal, family and social factors. In conclusion, prevention and protection should be reinforced. PMID- 20011905 TI - [Family and acquaintances of illicit drug users: community perspectives on laws and public policies in Western Rio de Janeiro, Brazil]. AB - This article is part of the study 'Illicit Drug Use in Seven Latin American Countries and Canada: Critical Perspectives of Family and Familiars' (7LACC), which investigated four domains: protective and risk factors; preventive initiatives; treatment facilities; and laws and policies. The article presents a section of the results based on four items of the laws and policies domain--as perceived by the family and acquaintances of illicit drug users living in the community. Participants were recruited in urban primary health care units located in Western Rio de Janeiro (city), Brazil. This multi-method, cross-temporal study performed interviews with 100 adults (18 years of age or older), all cognitively healthy. Results and key conclusions included non-compliance with the fundamental principles of the Unique Health System Legislation / Law 8.080/90 and the erroneous implementation of laws and public policies on illicit drug. PMID- 20011906 TI - [Laws and policies on illicit drugs in Brazil and the perspective of drug users' family members and acquaintances: a study in the City of Ribeirao Preto, Sao Paulo, Brazil]. AB - Brazilian drugs legislation has evolved from a prohibitionist system to a less repressive one in terms of drug users. The objective of this study was to identify the perception of relatives and acquaintances of drug users living in Ribeirao Preto, Sao Paulo, Brazil, about the country's laws and policies on drugs. Data collection was performed using a structured questionnaire. The sample consisted of 100 drug users' relatives or acquaintances, selected at a public health service. Respondents' relationships with the drug user were as follows: 31% friend, 23% sibling, 15% child and 7% spouse. Most users (78%) were men, with an average age of 26 years. Results confirm that national laws and policies have a direct effect on individuals' attitude and behaviors. There is a lack of trust in the police and a general perception that, despite recent chances that favor user rehabilitation, the laws on drugs do not respect users' human rights. PMID- 20011907 TI - [Illicit drug use and the critical perspectives of drug users' relatives and acquaintances in Northern Rio de Janeiro City, Brazil]. AB - This article presents the partial results of a multicenter, cross-temporal study, which was performed using multiple methods, and involved seven Latin-American countries and Canada. The results presented refer to the city center of Rio de Janeiro (n=108). The central question of the study was: 'How do illicit drug users' relatives and acquaintances describe protective and risk factors, prevention initiatives, treatment services, laws and policies regarding illicit drugs?' The quantitative data was collected using an instrument containing closed questions. In total, 108 young adults (18 years of age or older) were interviewed, who stated being affected by the drug although they were not users. For 104 interviewees (96%), negligence is the family dynamics that causes the greatest exposure to drugs, and 106 (98%) consider that parent support is what offers the greatest protection. Policies, the police and the criminal system have neither reduced drug use nor do they protect users. PMID- 20011908 TI - [Drug consumption and treatment from a family and friends perspectives: Guatemala]. AB - This quantitative and qualitative research describes the perspective of families and relatives of drug abusers in seven Latin American countries. In Guatemala, most of the people affected by the drug problem is multidrug abusers. Marijuana, followed by cocaine and benzodiazepines are the most used drugs. Of the respondents, 46% think drug use is a personal choice. They also recognize family as the most important protective factor, friends who use drugs and peer pressure are the major risk factors. The study reveals that the population believes that the response of the health services is insufficient, and that the preventive initiatives are not available or not properly addressed. The results show the need of more studies to update the knowledge of the drug problem in Guatemala. Future qualitative and quantitative research is needed to address the theme. PMID- 20011909 TI - [Critical perspective of relatives and acquaintances regarding protective factors and the use of illicit drugs in Guayaquil, Equador]. AB - The objective of this study was to determine the perspectives of drug users' relatives and acquaintances about protective factors for illicit drug use at a health center in Guayaquil. This is a descriptive, cross-sectional study. Data collection was performed through interviews using a questionnaire. Interviews were performed with 100 people who knew a drug user (relative or friend). The results showed that the following personal and family factors could be protective: 97% having solid moral principles, 96% express their feelings, 98% dedicate time for the family, and 95 % have a supportive relationship with one of the parents. Regarding the community, all participants (100%) agree there is a need for a government that understands this issue, 99% refer there should be honest policemen, and 99% state the need for programs that protect people from drug use and institutions that work with prevention. Family, community and personal decisions have effects on becoming involved, hence the need to reinforce protective factors and thus reduce the number of addicted individuals. PMID- 20011910 TI - University students' perceived norms of peers and drug use: a multicentric study in five Latin American countries. AB - This cross-sectional study compared perceived peer drug use and actual drug use in a sample of Latin American university students. Students from nine universities in five countries (Brazil, Chile, Colombia, Honduras and Peru) completed a questionnaire that addressed the use of tobacco, alcohol, marijuana and cocaine. Analysis focused on comparing perceptions to actual drug use. The findings largely, but not completely, confirmed the idea that students overestimate peer drug use. The unexpected findings were those relating to alcohol. While students generally overestimated peer use of tobacco, marijuana and cocaine, they accurately estimated or underestimated peer use of alcohol. Apart from the anomalous findings with regard to alcohol, this study shows that perceived drug use relates to actual drug use in Latin America as it does elsewhere. The results also support the suggestion that interventions using normative feedback would be useful to strengthen drug use prevention programs aimed at youth in Latin America. PMID- 20011911 TI - [Perceived norm for marijuana use among university student peers]. AB - This article describes the relationship between the perceived norm for marijuana use among peers and the use reported by university students in Chile. This is a qualitative, multicenter, cross-sectional study. A total 449 students participated with free consent. Data collection was performed using a self administered questionnaire, and SPSS 15.0 was used for data analysis. RESULTS: most participants were women, single, between 20 and 21 years of age, students of nursing and education. The perceived norm is higher than reports from national studies (32.1% against 21%) and higher than the self-reported use (32.1% against 5.6%). Marijuana use is higher among women, in nursing and education courses. It was confirmed that marijuana use among women begins at the university; as it was used as a recreational drug and there was more tolerance among peers. It was observed that marijuana use was overestimated, which, according to the Theory of Social Norms and Normalization, has an effect on the increase of drug use. This study is a contribution for the country's and the university's policies. PMID- 20011912 TI - [Perceived norms among Honduran university students about peers and tobacco, alcohol, marijuana and cocaine use]. AB - The purpose of this study was to assess the difference between perceived norms and peers' drug use among sophomore and junior university students (from the field of education) aged 18 to 24 years. The Social Norms Theory was used as the theoretical framework. In total, 286 students participated in the study, 67% of which reported having consumed alcohol at least once in a lifetime and 28% stated being daily users. Students perceived that 62% of their peers used tobacco and 63% used alcohol. The perceived norm for drug use was slightly higher in women than in men. In conclusion, there is an overestimation between the perceived norm and use of alcohol, tobacco, marijuana and cocaine. PMID- 20011913 TI - [Perceived norms among Peruvian students for drug use among peers]. AB - OBJECTIVE: to evaluate the difference between perceived norms about drug use among peer and actual drug use as reported by the same university students. The students were between 18 and 24 years old and attended health courses. METHOD: cross-sectional study based on a survey, using an anonymous questionnaire filled out by the students. RESULTS: There were 306 participants. Senior students used drugs as follows: 51.3% used tobacco, 90.8% used alcohol, 5.9% used marijuana, and 0.7% used cocaine. Differences were observed between perceived norms and actual drug use for tobacco (70% vs. 51.3%), marijuana (10% vs. 5.9%) and cocaine (8.3% vs. 0.7%). CONCLUSIONS: university students presented an overestimated rate for the use of tobacco, marijuana and cocaine among their peers. PMID- 20011914 TI - [Perceived norms among nursing students about their peers and drug use in Lima, Peru]. AB - This quantitative, cross-sectional study was performed to estimate the difference between the perceived norms among university students, based on a survey of 196 nursing students from Lima, Peru. A questionnaire was used to measure the students' perception about drug use and their actual use. Descriptive statistical analysis was performed, considering a value greater, smaller or equal to 10 to establish the difference. The perceived norms for alcohol and tobacco were above 50%, against 6% for marijuana and cocaine; with 57.7% of participants reporting tobacco use, 84.7% alcohol, 2.6% marijuana and 1% cocaine. Marijuana and cocaine use was overestimated, while alcohol use was underestimated. The perception of tobacco use was correct. The study findings are useful to sensitize university authorities and put prevention policies in practice. PMID- 20011915 TI - [University students' perception regarding drug use among peers in the ABC Region of Sao Paulo, Brazil]. AB - This study aimed to analyze the relation between university students' perceived norms and actual drug use among their peers. This cross-sectional study was based on a survey. Evaluations involved all second- and third-year students (ages between 18 and 24 years) of the nursing and medicine courses at the Faculdade de Medicina do ABC, located in the city of Santo Andre, Brazil. Students were invited to answer a questionnaire that evaluated several variables, including their own drug use and their perception of their peers' drug use. In total, 274 students participated. Data analysis was performed using SPSS and descriptive techniques, including frequencies and means. There was an overestimation of the perception regarding drug use in relation to the actual frequency reported by the students. PMID- 20011916 TI - [Perceived norms of peer alcohol use among university students]. AB - Alcohol is the most prevalent legal drug among university students, and it is used recreationally. The objectives of this study were to identify the perceived norms of alcohol use among university students, contrast the students' perception regarding peer alcohol use and the actual alcohol use prevalence, and compare the subjects' own alcohol use with their perception of peer alcohol use. A survey was performed on 275 students of a public university located in Rio de Janeiro (Brazil), in 2008. The students' perception about peer alcohol use once in a lifetime and over the last 12 years was correct; but their perception regarding the last 30 days was overestimated. Students consume alcoholic beverages recreationally, mainly at parties and bars with friends and peers. Alcohol use is above established standards. PMID- 20011918 TI - [Perceived norms among university students about their peers and drug use in Bogota, Colombia]. AB - The objective of this study was to estimate the difference between perceived norms for drug use among university students and their peers, with ages between 18 and 24 years, and attending education and health courses. This cross-sectional study was based on a survey and an anonymous self-administered questionnaire. RESULTS: in total, 365 students participated. The rates for drug use over the last year were as follows: 43.6% for tobacco; 96.2% for alcohol; 8.2% for marijuana; and 2.2% for cocaine. The perceived norm for drug use over the last year was 78.9% for tobacco, 88.3% for alcohol, 35.4% for marijuana and 20.9% for cocaine. The consumption of alcoholic beverages and the perceived norm were very close, but it was overestimated for tobacco use. Regarding access to drugs being easy or very easy on campus, access to cigarettes and cocaine was underestimated, while marijuana was overestimated. CONCLUSIONS: university students overestimate the use of tobacco, marijuana and cocaine by their peers and have a correct perception of alcohol use. Tobacco is the substance with the most overestimated use. PMID- 20011917 TI - [Consumption and perceived use of licit and illicit drugs among university students, Antioquia, Medellin, Colombia]. AB - The purpose of this study was to estimate differences between perceived and reported drug use among sophomore and junior university students, aged 18 to 24 years, from a public university in Medellin, Colombia. METHOD: this is a descriptive and cross-sectional study. All second- and third-year students at the Schools of Medicine, Dentistry, Nursing and Public Health were invited to answer an anonymous questionnaire. In total, 427 students participated. RESULTS: data suggest that students overestimate the use of tobacco, marijuana and cocaine over the last 12 months. Alcohol use was perceived accurately. Students who reported using those substances during that period overestimated their peers' drug use more than those who did not. Furthermore, more women than men overestimated marijuana use. PMID- 20011919 TI - [Perceived norms among university students of three health courses for drug use among peers]. AB - The present study interviewed second- and third-year students of three health courses to identify university students' estimates for drug use (perceived norm), the rate of drug use among the subjects (real norm); compare the estimates with the actual frequency; and identify in what conditions the drugs are used. Students overestimated their peers' use of tobacco, marijuana and cocaine in life and over the last 12 months. Alcohol was an exception. The rate values reported by sample students and the general estimated use were rather close. Drugs are usually consumed at parties and among friends from the university. Data analysis was performed in the light of the Social Norms Theory, Causal Attribution and Normalization. PMID- 20011920 TI - [Perceived norms among university students about their peers and drug use at a Chilean university]. AB - This study analyzes the relationship between perceived norms about drug use among peers and actual use among university students. This is a qualitative, multicenter study involving sophomore and junior students of the health courses of a Chilean university. Participants were 286 (75.2%) of the total 380 university students. RESULTS: more than 50% of the students notice that their peers smoke cigarettes on a daily basis, drink alcohol three times a week, and use marijuana once a week and cocaine at least once a year. The university campus it place of highest drug use; cocaine is used at a friend's house. Among students, 68% have smoked once in their life, 57% smoke on a daily basis; 88% have had alcohol once in their life; 26% used marijuana once in their life, 16% used marijuana over the last 12 months. Students do not know about any university policies for drug use. Some results confirm that drug use is underestimated. PMID- 20011922 TI - Quality of life, depression and anxiety among pregnant women with previous adverse pregnancy outcomes. AB - CONTEXT AND OBJECTIVE: Previous adverse pregnancy outcomes (recurrent spontaneous abortion, fetal death, preterm birth or early neonatal death) can affect the quality of life of pregnant women. The objective of this study was to compare the quality of life and the prevalence of symptoms of anxiety and depression among pregnant women with and without these antecedents. DESIGN AND SETTING: An analytical cross-sectional study was performed in four settings (two high-risk and two low-risk prenatal clinics) in the city of Campinas, Sao Paulo, Brazil. METHODS: A total of 240 women were interviewed by a single investigator between the 18th and 24th weeks of gestation: 120 women with prior adverse pregnancy outcomes (group 1) and 120 women with no such history (group 2), matched according to their numbers of living children. Sociodemographic variables were collected and two questionnaires were used: the Short Form-36 quality-of-life questionnaire and the Depression and Anxiety Scale. RESULTS: The women in group 1 had lower scores in all the items on the quality-of-life questionnaire. Depression and anxiety were more frequent in group 1 (P < 0.0001). An inverse correlation was found between the Short Form-36 domains and anxiety and depression. CONCLUSIONS: Women with histories of recurrent spontaneous abortion, fetal death, preterm birth or early neonatal death seem to have poorer quality of life and more symptoms of anxiety and depression during their subsequent pregnancy, compared with those without such antecedents. PMID- 20011923 TI - Approach towards total knee arthroplasty in Brazil: cross-sectional study. AB - CONTEXT AND OBJECTIVE: Total knee arthroplasty (TKA) has evolved particularly since the 1970s, with improvements in implants and surgical instruments, and has thus become an effective intervention for treating knee arthrosis. Many studies have presented rates of satisfactory clinical and radiological results greater than 90%, from follow-ups of over ten years. Nevertheless, despite scientific evidence showing the efficacy of TKA, the approaches taken present controversies in certain respects. The objective of this study was to evaluate how the Brazilian orthopedists deal with TKA, with investigation of the main aspects of this procedure. DESIGN AND SETTING: Cross-sectional survey conducted during the 39th Brazilian Congress of Orthopedics and Traumatology, in Sao Paulo, Brazil, in November 2007. METHODS: We applied a questionnaire to orthopedists registered at the congress. The questionnaire was randomly distributed and participation was voluntary; 858 completed questionnaires were included in the analysis. RESULTS: Most of the Brazilian orthopedists were members of SBOT and worked in the southeastern region. They used imported cemented implants through an anterior access route centered on the patella, with replacement of the joint surface of the patella and preservation of the posterior cruciate ligament. They did not have experience with simultaneous bilateral TKA. Postoperatively, they used antibiotics and suction drains for 48 hours. There was no consensus regarding prophylaxis for venous thromboembolism or the frequency of the main complications. CONCLUSION: The majority of Brazilian orthopedists work in the southeastern region of the country and agree about the main aspects of the approaches towards TKA. PMID- 20011924 TI - Cross-sectional study on the weight and length of infants in the interior of the state of Sao Paulo, Brazil: associations with sociodemographic variables and breastfeeding. AB - CONTEXT AND OBJECTIVE: Increasing obesity is starting to occur among Brazilians. The aim of this study was to investigate the weight and length of children under two years of age in relation to sociodemographic variables and according to whether they were breastfed. DESIGN AND SETTING: Cross-sectional randomized study conducted in 2004-2005, based on the declaration of live births (SINASC) in Campinas, Brazil. METHODS: 2,857 mothers of newborns were interviewed and answered a questionnaire seeking socioeconomic and breastfeeding information. The newborns' weights and lengths were measured at the end of the interviews and the body mass index was calculated. Percentiles (< 15 and > 85) and Z-scores (< -1 and > +1) were used for classification based on the new growth charts recommended by WHO (2006). The log-rank test, multiple linear regression and binomial test (Z) were used. The statistical significance level used was 5%. RESULTS: The predominant social level was class C. The median for exclusive breastfeeding was 90 days; 61.25% of the children were between P15 and P85 for body mass index and 61.12% for length, respectively. Children whose mothers studied for nine to eleven years and children whose mothers were unemployed presented lower weight. Children whose mothers worked in health-related professions presented lower length when correlated with breastfeeding. CONCLUSION: The breastfeeding, maternal schooling and maternal occupation levels had an influence on nutrition status and indicated that obesity is occurring in early childhood among the infants living in the municipality. PMID- 20011925 TI - Frequency of potential interactions between drugs in medical prescriptions in a city in southern Brazil. AB - CONTEXT AND OBJECTIVE: Drug interactions form part of current clinical practice and they affect between 3 and 5% of polypharmacy patients. The aim of this study was to identify the frequency of potential drug-drug interactions in prescriptions for adult and elderly patients. TYPE OF STUDY AND SETTING: Cross sectional pharmacoepidemiological survey in the Parque Verde housing project, municipality of Cascavel, Parana, Brazil, between December 2006 and February 2007. METHODS: Stratified cluster sampling, proportional to the total number of homes in the housing project, was used. The sample consisted of 95 homes and 96 male or female patients aged 19 or over, with medical prescriptions for at least two pharmaceutical drugs. Interactions were identified using DrugDigest, Medscape and Micromedex softwares. RESULTS: Most of the patients were female (69.8%), married (59.4%) and in the age group of 60 years or over (56.3%), with an income less than or equal to three minimum monthly salaries (81.3%) and less than eight years of schooling (69.8%); 90.6% of the patients were living with another person. The total number of pharmaceutical drugs was 406 (average of 4.2 medications per patient). The drugs most prescribed were antihypertensives (47.5%). The frequency of drug interactions was 66.6%. Among the 154 potential drug interactions, 4.6% were classified as major, 65.6% as moderate and 20.1% as minor. CONCLUSION: The high frequency of drug prescriptions with a potential for differentiated interactions indicates a situation that has so far been little explored, albeit a reality in household surveys. PMID- 20011926 TI - Prevalence of pressure ulcers among the elderly living in long-stay institutions in Sao Paulo. AB - CONTEXT AND OBJECTIVE: The prevalence of pressure ulcers varies according to geographic region and population group, such as the institutionalized elderly. The aim of this study was to identify the prevalence of pressure ulcers among elderly people living in long-stay institutions. DESIGN AND SETTING: Cross sectional study in six long-stay institutions for the elderly in Sao Paulo, Brazil. METHODS: Demographic and clinical data were collected in six long-stay institutions on two visits to each institution between May and August 2007, during which all elderly patients with pressure ulcers were evaluated. The Braden scale was used to identify the risk of developing pressure ulcers and the National Pressure Ulcer Advisory Panel (NPUAP) stages for classifying the pressure ulcers. Statistical analysis was performed using the chi-square test, Student's t-test and Fisher's exact test. RESULTS: There was no significant difference in the results between visits. The population was 181 elderly people in May and 184 in August: 23 had pressure ulcers in May (prevalence of 12.7%) and 17 in August (prevalence of 9.2%). The mean age at the two times was 84 years, and the average length of stay was 32 months. Pressure ulcers were found mainly in the sacral region (mean, 71.5%), and most commonly in stage II (mean, 41%). CONCLUSION: The prevalence of pressure ulcers was 10.95%. These data provide background information that may aid in developing protocols for applying best practices for prevention and treatment of pressure ulcers, consequently reducing the prevalence. PMID- 20011927 TI - Association between osteoporosis and rheumatoid arthritis in women: a cross sectional study. AB - CONTEXT AND OBJECTIVES: Osteoporosis has frequently been observed in patients with rheumatoid arthritis. The present study was undertaken in order to evaluate factors associated with osteoporosis among women with rheumatoid arthritis. DESIGN AND SETTING: Cross-sectional study, carried out in a public hospital in Sao Paulo. METHODS: The participants were 83 women with rheumatoid arthritis (53.7 +/- 10.0 years old). Bone mineral density (BMD) and body composition were measured by dual energy X-ray absorptiometry. The patients were divided into three groups according to BMD: group 1, normal BMD (n = 24); group 2, osteopenia (n = 38); and group 3, osteoporosis (n = 21). Tests were performed to compare differences in means and correlations, with adjustments for age, duration of disease and cumulative corticosteroid. The relationships between clinical factors, physical activity score, dietary intake, body composition and biochemical parameters were analyzed using linear regression models. RESULTS: Mean calcium, vitamin D and omega-6 intakes were lower than the recommendations. Associations were found between BMD and age, disease duration, parathyroid hormone concentration and fat intake. The linear regression model showed that being older, with more years of disease and lower weight were negatively correlated with BMD [Total femur = 0.552 + 0.06 (weight) + 0.019 (total physical activity) - 0.05 (age) - 0.003 (disease duration); R(2) = 48.1; P < 0.001]. CONCLUSION: The present study indicates that nutritional factors and body composition are associated with bone mass in women with rheumatoid arthritis. PMID- 20011928 TI - Hepatopulmonary syndrome: an update. AB - Hepatopulmonary syndrome (HPS) is a clinical threesome composed of liver disease, intrapulmonary vascular dilatation (IPVD) and arterial gas abnormalities. Its occurrence has been described in up to 32% of cirrhotic candidates for liver transplantation. It also affects non-cirrhotic patients with portal hypertension. Its pathogenesis is not well defined, but an association of factors such as imbalance in the endothelin receptor response, pulmonary microvascular remodeling and genetic predisposition is thought to lead to IPVD. Diagnosis is based on imaging methods that identify these dilatations, such as contrast echocardiography or perfusion scintigraphy with 99mTc, as well as analysis of arterial gases to identify elevated alveolar-arterial differences in O2 or hypoxemia. There is no effective pharmacological treatment and complete resolution only occurs through liver transplantation. The importance of diagnosing HPS lies in prioritizing transplant candidates, since presence of HPS is associated with worse prognosis. The aim of this paper was to review the pathogenetic theories and current diagnostic criteria regarding HPS, and to critically analyze the prioritization of patients with HPS on the liver transplant waiting list. Searches were carried out in the Medline (Medical Literature Analysis and Retrieval System Online) via PubMed, Cochrane Library and Lilacs (Literatura Latino-Americana e do Caribe em Ciencias da Saude) databases for articles published between January 2002 and December 2007 involving adults and written either in English or in Portuguese, using the term hepatopulmonary syndrome. The studies of greatest relevance were included in the review, along with text books and articles cited in references that were obtained through the review. PMID- 20011929 TI - The role of perioperative warming in surgery: a systematic review. AB - OBJECTIVE: The objective of this review was to systematically analyze the trials on the effectiveness of perioperative warming in surgical patients. METHODS: A systematic review of the literature was undertaken. Clinical trials on perioperative warming were selected according to specific criteria and analyzed to generate summative data expressed as standardized mean difference (SMD). RESULTS: Twenty-five studies encompassing 3,599 patients in various surgical disciplines were retrieved from the electronic databases. Nineteen randomized trials on 1785 patients qualified for this review. The no-warming group developed statistically significant hypothermia. In the fixed effect model, the warming group had significantly less pain and lower incidence of wound infection, compared with the no-warming group. In the random effect model, the warming group was also associated with lower risk of post-anesthetic shivering. Both in the random and the fixed effect models, the warming group was associated with significantly less blood loss. However, there was significant heterogeneity among the trials. CONCLUSION: Perioperative warming of surgical patients is effective in reducing postoperative wound pain, wound infection and shivering. Systemic warming of the surgical patient is also associated with less perioperative blood loss through preventing hypothermia-induced coagulopathy. Perioperative warming may be given routinely to all patients of various surgical disciplines in order to counteract the consequences of hypothermia. PMID- 20011930 TI - Ureteral fibroepithelial polyps in a pregnant woman: case report. AB - CONTEXT: Ureteral fibroepithelial polyps are rare benign nonepithelial tumors, and less than 200 cases have been reported in the literature. We report on a pregnant patient with ureteral fibroepithelial polyps that were successfully treated with laparotomy. CASE REPORT: A 23-year-old pregnant woman presented with a three-month history of intermittent lumbar pain of low intensity. Abdominal ultrasonography showed that she was 13 weeks pregnant and found severe left-side ureterohydronephrosis and a heterogeneous solid mass measuring 11 x 8 x 7 centimeters in the middle portion of the ureteral topography. The investigation was complemented with magnetic resonance imaging, which confirmed the previous findings. Nephroureterectomy was performed without complications. The specimen revealed three solid tumors in the ureter, of which the largest was around eight centimeters in length. The anatomopathological report confirmed that they were fibroepithelial tumors without malignant components. PMID- 20011931 TI - Cissampelos sympodialis Eichl (Menispermaceae) leaf extract as a possible novel and safe treatment for psoriasis. PMID- 20011933 TI - [Estimation of coronary heart disease risk in Chilean subjects based on adapted Framingham equations]. AB - BACKGROUND: Cardiovascular disease is the leading cause of morbidity and mortality in Chile and worldwide. Framingham functions were developed to calculate overall coronary heart disease risk. However these functions overestimate the risk in some countries like Chile. AIM: To develop Chilean risk tables to assess the overall 10-year risk of coronary heart disease. MATERIAL AND METHODS: The Framingham function was adapted for a population aged 35 to 74 years, based on an estimate of Chilean incidence of coronary heart disease and the prevalence of coronary heart disease risk factors such as age, sex, total cholesterol, high-density lipoprotein cholesterol, blood pressure, diabetes and smoking. RESULTS: The 10-year incidence of coronary heart disease in Chile (2.7% in men, 1.096 in women) was lower than the incidence in the United States (10.096/3.896) and Spain (4.996/2.296), but higher than China (1.196/0.496). Framingham tables have more than 50% of cells in the risk category of 10% or greater. In contrast, Chilean tables have less than 10% of cells in the same risk category. CONCLUSIONS: Adapted tables use local information to calculate overall coronary heart disease risk. A validation study should be conducted to assess their predictive power. PMID- 20011934 TI - [Management of community acquired pneumonia in adults following clinical guidelines at a rural hospital]. AB - BACKGROUND: A National Consensus Guideline published in 2005 established the basis for the diagnostic, severity assessment and treatment of community acquired pneumonia (CAP) in the adult population. The compliance with pneumonia clinical guidelines has been associated to a reduction in hospital stay healthcare-related costs, morbidity and mortality. AIM: To describe the management and outcome of non-severe CAP in hospitalized adult patients treated in a rural hospital, based on the national clinical guidelines. PATIENTS AND METHODS: Ninety six patients aged 74 +/- 13 years (50 males) hospitalized with non-severe pneumonia (group 3) at a community-based primary care center between January 1, 2006, and March 31, 2007, were evaluated. RESULTS: Eighty percent of patients had concomitant diseases such as hypertension in 49%, diabetes in 23% and chronic obstructive pulmonary disease in 18%. All were treated with a third generation cephalosporin (ceftriaxone 1-2 g/day TV) as empirical therapy. Only 9% of patients also received a macrolide. Early switch to oral antimicrobial therapy was successful in two third of cases. Mean hospital length of stay was 5.0 +/- 2.5 days, and 30 day mortality was 6.3%. CONCLUSIONS: Following the recommendations of the national clinical guidelines, most of these patients had a favorable response to monotherapy with a beta-lactam antimicrobial. Early switch therapy to oral antibiotic was effective and safe, reducing significantly hospital length of stay as compared to previous national clinical studies. PMID- 20011935 TI - [Modeling the academic performance of medical students in basic sciences and pre clinical courses: a longitudinal study]. AB - BACKGROUND: The study of predictors of academic performance is relevant for medical education. Most studies of academic performance use global ratings as outcome measure, and do not evaluate the influence of the assessment methods. AIM: To model by multivariate analysis, the academic performance of medical considering, besides academic and demographic variables, the methods used to assess students' learning and their preferred modes of information processing. MATERIAL AND METHODS: Two hundred seventy two students admitted to the medical school of the Pontificia Universidad Catolica de Chile from 2000 to 2003. Six groups of variables were studied to model the students' performance in five basic science courses (Anatomy, Biology, Calculus, Chemistry and Physics) and two pre clinical courses (Integrated Medical Clinic I and IT). The assessment methods examined were multiple choice question tests, Objective Structured Clinical Examination and tutor appraisal. RESULTS: The results of the university admission tests (high school grades, mathematics and biology tests), the assessment methods used, the curricular year and previous application to medical school, were predictors of academic performance. The information processing modes influenced academic performance, but only in interaction with other variables. Perception (abstract or concrete) interacted with the assessment methods, and information use (active or reflexive), with sex. The correlation between the real and predicted grades was 0.7. CONCLUSIONS: In addition to the academic results obtained prior to university entrance, the methods of assessment used in the university and the information processing modes influence the academic performance of medical students in basic and preclinical courses. PMID- 20011936 TI - [Thelarche and nutritional status: an epidemiological study of two ethnic groups]. AB - BACKGROUND: Improvements in environmental conditions may result in an earlier onset of thelarche. However, its onset is not homogeneous among different population groups. AIM: To assess the relationship between nutritional status, ethnicity and age of thelarche. MATERIAL AND METHODS: Cross-sectional study of girls in second through sixth grade, attending 165 schools located in Chile's Araucania region. Of these, 231 girls who presented thelarche (breast button) were selected. The girls' surnames were used to identify their ethnicity: indigenous had three or four last names of Mapuche indigenous origin (n =113), while non-indigenous were those who only had Chilean-Spanish last names (n =118). Weight, height, waist circumference and skinfold thicknesses were measured and socio-economic background information was collected through a home interview. RESULTS: The median age of thelarche was 10 years and 4 months, regardless of ethnic group. Multivariant models showed that an increase of one z score unit of body mass index (BMI) decreased the median age of thelarche by 5.6 months (CI: 7.24 to -3.90), controlling for the effect of different covariables. A decrease in one z score of height retards the median age of thelarche by 5.5 months (CI: 4.02 to 6.98). Ethnicity did not influence the age of thelarche. CONCLUSIONS: The age of thelarche found by us is similar to that reported in international studies, it comes earlier as weight increases, is delayed as height decreases and is not related to ethnicity. PMID- 20011937 TI - [Antimicrobial susceptibility of Helicobacter pylori strains isolated in Colombia]. AB - BACKGROUND: Helicobacter pylori antimicrobial resistance rates differ among countries and even between different areas of a country. In Colombia, the most commonly used antimicrobials for the treatment of H pylori infection are amoxicillin, clarithromycin and metronidazole. AIM: To determine antimicrobial susceptibility of H pylori strains isolated in Colombia. MATERIALS AND METHODS: Eighty eight strains of H pylori were isolated and identified by microbiological methods and confirmed with polymerase chain reaction (PCR). The detection of antimicrobial resistance to amoxicillin, clarithromycin, metronidazole and tetraclycline, was conducted by the Etest method. Mutations in the 23S rDNA, involved in resistance to clarithromycin, were detected using PCR and restriction fragment length polymorphism. RESULTS: Eighty eight and 2.2% of the strains were resistant to metronidazole and clarithromycin, respectively. No isolate was simultaneously resistant to amoxicillin or tetracycline. The two clarithromycin resistant strains were homozygous for the A2143G mutation. No mutations were found in the remaining 86 susceptible strains. CONCLUSIONS: The high rate of metronidazole resistance in our population precludes the use of this drug for the empirical treatment of H pylori infection. PMID- 20011938 TI - [Effects of periodontal therapy on markers of systemic inflammation in patients with coronary heart disease risk]. AB - BACKGROUND: Studies investigating effects of periodontal treatment (PT) on markers of inflammation in healthy subjects show conflicting results. Few studies have investigated the effects of PT among subjects with coronary heart disease (CHD) risk factors. AIM: To report the results of a pilot prospective study on the effects of periodontal treatment on markers of inflammation among subjects with CHD risk factors. MATERIAL AND METHODS: Seventy three patients aged 53+/-6 years (25% males) with chronic periodontitis, dyslipidemia and other CHD risk factors were subjected to PT consisting on root planning and oral metronidazol and amoxicillin for 7 days. Periodontal clinical parameters, serum C-reactive protein (CRP), fibrinogen levels and erythrocyte sedimentation rate (ESR) were assessed before and at 6 weeks after PT. Polymorphisms at the ILlA-889 and IL1B+3954 genes were also genotyped. RESULTS: After the treatment period, CRP levels significantly increased from 3.6+/-3.7 mg/ L to 5.4+/-5.7 mg/L (p =0.001). No significant changes were observed in fibrinogen levels and ESR. Higher post treatment CRP levels were significantly associated with the composite polymorphic genotype at the ILlA-889 and IL1B+3954 genes (p =0.0001), and extensive periodontitis (p =0.005). Moderate alcohol consumption appeared as a protective factor for CRP elevation (p =0.029). CONCLUSIONS: The increase of the CRP levels after PT in patients with CVD risk factors appeared associated with IL-1 gene polymorphisms and extensive periodontitis. PMID- 20011939 TI - [Effects of incorporating group visits on the metabolic control of type 2 diabetic patients]. AB - BACKGROUND: A better metabolic control is an important objective of health care in diabetes mellitus. This objective has been achieved elsewhere, incorporating group visits to the usual care of diabetic patients. AIM: To evaluate the effects of a primary care group visit model after 9 and 15 months of implementation, on the metabolic control of diabetic patients. MATERIAL AND METHODS: Two cohorts of type 2 diabetic patients, matched by sex, age and fasting plasma glucose (FPG) were studied. Group visits (monthly medical consultation and one hour sessions with group interaction on the same day) were implemented in 600 patients and routine visits (monthly one-to-one medical consultation) were implemented in 601 patients. The evolution on FPG, cholesterol, systolic (SBP) and diastolic blood pressure (DBF) and body mass index (BMI) were compared. RESULTS: At 15 months of follow up, mean FPG was lower in patients with group visits compared to those in usual individual care (155.3 +/- 59.5 and 175.7 +/- 67.7 mg/dL respectively, p <0.01). SBP and DBF were also lower in patients on group visits (123.6 +/- 13.4 and 127.5 +/-12.8 mmHg, respectively for systolic pressure, p <0.01 and 73.5 +/ 8.5 and 79.4 +/-6.3 mmHg, respectively for diastolic pressure, p <0.01). No differences between groups were observed for blood cholesterol or BMI. CONCLUSIONS: Incorporation of group visits in type 2 diabetic patients improved metabolic control and blood pressure, compared to the usual individual care model. PMID- 20011940 TI - [Retrospective review of 44 Chilean patients with Behcet disease]. AB - BACKGROUND: Behcet's disease (BD) is a rare multisystemic inflammatory disease that is potentially disabling and may cause death. AIM: To describe the characteristics of BD patients from two Chilean centers. PATIENTS AND METHOD: Retrospective review of the clinical records of patients with BD attended in two rheumatology services between 1985 and 2007. The "Behcet's Disease Research Committee of Japan" (BDCJ) and the "International Study Group for Behcet's Disease" (ISG) diagnostic criteria were applied. RESULTS: We found 44 cases (25 males), diagnosed as BD. The mean age at the onset of symptoms was 26+/- 12 years. According to BDCJ criteria, 13 patients had complete BD, 24 had incomplete BD and 7 had a suspected BD. Thirty two patients fulfilled the ISG criteria. Forty two patients (95%) had oral ulcers, 33 (75%) had genital ulcers and 29 (66%) had ophthalmological involvement. Eleven and three patients had symptoms of central and peripheral nervous system involvement, respectively. No gender differences were detected. CONCLUSIONS: The clinical characteristics of these patients were similar to those described abroad, except for a higher frequency of peripheral nervous system involvement and a lower rate of arthritis. PMID- 20011941 TI - [Adenocarcinoma and intestinal duplication of the ileum. Report of one case]. AB - Gastrointestinal duplications are uncommon developmental abnormalities that occur anywhere along the gastrointestinal tract. We report a 36 year-old female admitted to the emergency room due to abdominal pain. Computed abdominal tomography demonstrated small bowel loops surrounding a tubular cystic structure and peritoneal free fluid. The surgical exploration revealed multiple malignant implants covering the visceral and parietal peritoneum and infiltrating completely the omentum. At the ileal mesentery we found a tubular cystic whitish tumor measuring 12 cm of diameter and 15 cm on length. A complete resection of the tumor was not considered an option due to the extensive peritoneal dissemination. Thirty-four days after the operation the patient died. The histopathology of the cystic wall was compatible with the architecture of intestinal wall extensively infiltrated by a moderately differentiated mucinous adenocarcinoma; a mucosal lining in parts atrophic and in parts infiltrated or replaced by adenocarcinoma was observed. A well structured muscular layer was recognized, and the myenteric plexus was identified. PMID- 20011942 TI - [Complete repair in a single period of circulatory arrest of a ruptured atherosclerotic aortic arch aneurysm. Report of one case]. AB - The aortic arch aneurysm is a complex disease with a growing incidence in our population. We report a 64 year-old female with an atherosclerotic aortic arch aneurysm with a maximum diameter of 9.4 cm and evidence of contained rupture. We performed a complete aortic arch replacement in a single period of 100 minutes of circulatory arrest with deep hypothermia and brain protection with anterograde cerebral perfusion through the right axillary artery. The patient had a complete recovery without evidence of neurological damage and was extubated on the next morning. One year later, the patient is asymptomatic and the peri-aortic haematoma was completely reabsorbed. PMID- 20011943 TI - [Prolonged prone position ventilation for severe respiratory distress syndrome post-pneumonectomy. Report of one case]. AB - Management of patients with severe respiratory failure is mainly supportive, and protective mechanical ventilation is the pivotal treatment. When conventional therapy is insufficient to improve oxygenation without deleterious effects, other strategies should be considered. We report a 53 year-old male who presented a severe respiratory failure refractory to conventional management after pneumonectomy. Prone position ventilation was used for 36 hours. Respiratory variables improved and he did not show hemodynamic instability. He was returned to the supine position without worsening of oxygenation parameters. Extended prone position ventilation could be considered in patients presenting with unresponsive severe respiratory failure after pulmonary resection. PMID- 20011944 TI - [Cardiogenic shock secondary to acute myocardial infarction managed with high volume hemofiltration: report of a case]. AB - Cardiogenic shock secondary to acute myocardial infarction unveils a systemic inflammatory response with elevation of cytokines that contribute to hypoperfusion. High volume hemofiltration may remove cytokines in patients with septic shock resulting in hemodynamic improvement and reducing the requirements of norepinephrine. We report a 48 year-old male with cardiogenic shock secondary to acute myocardial infarction who presented a systemic inflammatory response characterized by fever and hemodynamic collapse, without evidence of infection. Its hemodynamic profile was stabilized with high volume hemofiltration. PMID- 20011945 TI - [Bridge to lung transplantation with a novel pumpless lung assist device. Report of one case]. AB - Devices such as Novalung can be used as a bridge to lung transplantation while waiting for a suitable donor. We report a 50 year-old male with a terminal pulmonary fibrosis and candidate for lung transplantation. He was admitted to the hospital due to a severe deterioration of his respiratory condition, with the presence of severe respiratory acidosis despite conventional invasive respiratory support. Respiratory support with Novalung was started, resulting in a progressive reduction of pCO2 that became normal ten hours after the installation of the device. Five days later a successful lung transplantation was performed. PMID- 20011946 TI - [Infections in patients with Systemic Lupus Erythematosus]. AB - Despite the availability of newer therapeutic interventions to improve clinical outcome in patients with Systemic Lupus Erythematosus (SLE), the incidence of infections as a cause of morbidity and mortality has not changed over the past 30 years. SLE itself increases the risk of infection, due to genetic (complement deficiencies) and acquired factors such as functional asplenia (humoral immunodepression) and the use of immunosuppressive drugs. These medications increase the risk of opportunistic infections that are associated with an altered cellular immune response. The main etiologic infectious agents in SLE patients are common bacterial pathogens, especially capsulated ones. The most common sites are lung, skin, bladder, brain and systemic infections. The main risk factor for infection is the history of a previous one. The clinical approach to SLE patients with suspected infectious diseases must consider the possibility of a flare up of the underlying disease, posing an additional problem to the clinician. PMID- 20011947 TI - [Pathophysiology of diabetic retinopathy and nephropathy]. AB - Despite the availability of multiple therapeutic approaches, diabetes mellitus with chronic hyperglycemia remains as the main cause of new cases of blindness and chronic renal failure in the western hemisphere. We herein review the molecular mechanisms by which chronic hyperglycemia causes retinopathy and nephropathy in type I and type 2 diabetic patients. Diabetic retinopathy develops silently along years or decades, producing symptoms only in its very late stages. Its slow development starts with the activation of aldose reductase, shortly followed by the destruction of the retinal pericyte cells, and ends in sudden blindness when vitreous hemorrhage ensues. Nephropathy, on the other hand, centers its pathophysiology in the mesangial cell, that starts as a modified smooth-muscle cell, and turns itself into a myo-fibroblast, produces such amounts of cytoplasm and extracellular protein that strangulates the glomerular capillaries and causes renal failure. After a detailed review of the molecular mechanisms of the aforementioned complications, we conclude that, apart from directing our attention to the emerging medications that are being developed to block these molecular pathways, we should never abandon the struggle for improving the glycemic control of our diabetic patients. PMID- 20011948 TI - [Hospital medicine]. AB - Hospital medicine was created over 10 years ago aiming to provide an integral care to hospitalized patients. Hospital specialists are physicians mainly devoted to the global care of hospitalized patients. Their professional functions include patient care, teaching, clinical research and managing activities. The main difference with other specialties is their exclusive dedication to hospital work. The impact of this specialty on patient care has been demonstrated by a significant reduction in the hospitalization days and costs and higher level of patient satisfaction. In clinical hospitals, the presence of tutors during the complete working day, has resulted in better pre and postgraduate teaching activities and a higher availability of supervisors for trainees. Four years ago, hospital medicine was established as a discipline at the Clinical Hospital of Pontificia Universidad Catolica de Chile. In this period, these specialists became essential for student training and an integral part of the faculty staff. PMID- 20011949 TI - [Benefits from the exercise of autonomy and informed consent: the example of Jehovah's Witnesses]. AB - The respect for self-determination has represented a great challenge for the medical community. This debate has resulted in laws, codes of ethics, international treaties, and administrative guidelines, all with the purpose of protecting such right. In the medical field, the "Convention for the Protection of Human Rights and Dignity of the Human Being with Regard to the Application of Biology and Medicine: Convention on Human Rights and Biomedicine", known as the Oviedo Convention plays a crucial role. The doctrine of Informed Consent (IC) exists to enforce it. This principle is considered as law in some countries. In Chile, the IC is considered in the legal ordinance and in documents that are used as guidelines in the field of ethics. Jehovah s Witnesses invoke such precepts when they demand respect for their decisions. The present article outlines their position regarding blood transfusions and their contribution to the practice of bloodless medicine and surgery, which promotes the respect for patient's self determination. The experience of Jehovah's Witnesses has lead to a dignified treatment of the patient and has promoted a better participation in decision making, focusing on the patient. All these benefits can be conveyed to other patients, resulting in the protection of the dignity of the individual. PMID- 20011950 TI - [Clinical usefulness of toxicology testing]. AB - A toxicology testing is the search by the laboratory of the possible etiologic agents that can cause poisoning. Given the wide variety of substances that can poison a person, the laboratories should work coordinated with the emergency wards in order to determine the appropriate tests menu and the required turn around time according to the most frequent causes of intoxication in the local population. Toxicology laboratories should provide two tiers of drug testing: selected drug tests in blood/urine and comprehensive or broad-spectrum toxicological testing in the same or other samples. The medical order must always include the suspected diagnosis, which is responsibility of the physician requesting the test. A most important issue in the study of a poisoned patient is the opportunity when the samples are drawn, which should be at the emergency room since a delay in sample collection implies losing unrecoverable information. Samples should be sent to the laboratory for either immediate analysis or later comprehensive toxicological tests, so that laboratories must have procedures for the proper storage and preservation of samples. Poison control centers provide assistance to clinicians in considering certain drugs etiologies and in selecting specific tests. PMID- 20011951 TI - [Smoking and alcohol drinking among medical students]. PMID- 20011952 TI - [Acute lobar nephronia]. PMID- 20011953 TI - [Laboratory alert value reporting by the clinical laboratory at an academic medical network]. AB - BACKGROUND: An alert value is a result suggesting that the patient is at imminent danger unless appropriate remedial actions begin promptly. Report of alert values (AV) by the clinical laboratories has taken special relevance in recent years due to its contribution to patient's care. AIM: To report results of AV informed during 2007 within the Health Network of the Pontificia Universidad Catolica de Chile. MATERIAL AND METHODS: Analysis of AV recorded in a centralized database of the laboratories of the health network, between January and December, 2007. RESULTS: Total number of AV was 5.366, which represented 0.3% of total examinations and corresponded mainly to the clinical chemistry area. Potassium levels generated the higher number of AV detected, followed by positive blood cultures. Eighty two percent of AV corresponded to hospitalized patients. The greater number of AV was reported to intermediate and intensive care services. Thirty two percent of AV was informed to the physician or professional in charge of the patient within 5 minutes of obtaining the results and 79% within 30 minutes. CONCLUSIONS: To obtain a real impact on patient management, it is fundamental to shorten the lapse between the obtainment of tests results and the warning, supported on appropriate computerized systems, and to spread the procedure to all personnel involved in patient's care. PMID- 20011954 TI - [Natural history of diabetic retinopathy in a retrospective cohort of type 1 diabetics]. AB - BACKGROUND: Diabetic retinopathy is one of the most common causes of blindness among adults. AIM: To report the natural history of diabetic retinopathy among Chilean patients with type 1 diabetes followed for a mean of 18 years. MATERIAL AND METHODS: Retrospective review of medical records of 39 patients aged 26 to 70 years, (20 females, 78 eyes) with type 1 diabetes controlled by the same ophthalmologist from 1971 to 2008. A questionnaire was sent to each patient and their treating physician to request information about the evolution of the disease and metabolic control. RESULTS: The questionnaire was answered by 24 patients (62%) and 21 attending physicians (54%). Small hard drusen were observed in 25 patients (64%). In 12 cases the drusen were detected before the development of any type of retinopathy. Eleven women became pregnant and retinopathy progressed in four of them. Twenty three patients (59%) developed proliferative diabetic retinopathy (PDR). Patients with PDR had a significantly longer duration of diabetes and worse glycemic control. There was a higher frequency of diabetic nephropathy in the PDR group, but only 13 patients out of 23 with PDR had nephropathy. The retinopathy progressed to high risk PDR two years after successful kidney-pancreas transplantation in one patient. CONCLUSIONS: In patients with type 1 diabetes mellitus, small hard drusen may be the initial manifestation of diabetic retinopathy. Risk factors for progression to PDR were duration of diabetic and poor glycemic control. Nephropathy was more prevalent in patients with PDR, but a significant group of PDR patients did not have demonstrable nephropathy. PMID- 20011955 TI - [Early and late morbidity and mortality of aortic valve replacement with mechanical or biological prostheses during the last decade: the gold standard]. AB - BACKGROUND: There is a growing interest in alternative techniques for aortic valve replacement (AVR). Therefore it is important to have updated results of conventional AYR as a valid comparative standard. AIM: To evaluate both perioperative and late morbidity and mortality in patients undergoing conventional AVR, with either mechanical (RVAm) or biological (RVAb) prostheses. PATIENTS AND METHODS: Retrospective review of medical records and operative protocols of patients undergoing AVR between January 1995 and December 2005. Patients with previous cardiac surgery, aortic balloonplasty or simultaneous cardiovascular procedures were excluded. RESULTS: During the study period, 788 patients underwent AVR and 317 met the inclusion criteria. Of the latter, 175 patients aged 13 to 83 years (63% males) were subjected to AVRm and 142 patients aged 49 to 87 years (64% males), were subjected to AVRb. Five (1.6%) patients died during the perioperative period (one AVRm and four AVRb). All were older than 65 years. Perioperative complications were recorded in 29 and 25% of patients in AVRm and AVRb groups, respectively (NS). During a median follow-up of six years, complications were recorded in 12 and 4% of patients in AVRm and AVRb groups, respectively (p <0.05). The actuarial survival for AVRm group at 1, 5 and 10 years, was 96, 92 and 87%, respectively. The figures for AVRb group were 95, 86 and 83%, respectively (NS). The 10 year reintervention free survival was 97% for the AVRm group and 84% for the AVRb (p <0.05). CONCLUSIONS: Perioperative mortality and rates of complications of AVR in this series of patients are low, which compares favorably with other series. PMID- 20011956 TI - [Factor analysis of health related quality of life in patients admitted to a short stay medical unit]. AB - BACKGROUND: Disease and hospital admission have a great impact on the quality of life of an individual. AIM: To identify the factors associated with health related quality of life (HRQL) in patients admitted to a Short Stay Medical Unit (SSMU). PATIENTS AND METHODS: Prospective cohort study of 335 patients aged 15 to 99 years (196 males) consecutively admitted to the SSMU. In all we recorded sociodemographic data, number of previous hospital admissions, admission type, main diagnosis, lenght of stay, comorbidities, the Goldberg Depression and Anxiety Scale, the Karnofsky Perfomance Scale, the Barthel Index and quality of life with the EuroQol-5D instrument. After studying the associations between variables, we carried out a factor analysis of those that were significantly related to HRQL. RESULTS: Mean body mass index was 27.7 Kg/m(2) and 83% were emergency admissions. Heart failure, chronic obstructive pulmonary disease exacerbation, acute coronary syndromes and cardiac arrhythmias, constituted 56% of all admissions. HRQL was statistically related to age, gender, educational status, caregiving situation, number of previous admissions, main diagnosis, length of stay, Goldberg scale and Karnofsky, Barthel and Charlson indexes. Factor analysis reduced the original variables to five, which explained 67.8% of the variance, as follows: Factor 1- Karnofsky and Barthel indexes (27.8%); Factor 2- age, educational status, caregiving situation (12.3%); Factor 3- the Goldberg scale (10.4%); Factor 4- admission type (8.8%); Factor 5- main diagnosis (8.4%). CONCLUSIONS: HRQL in patients admitted to Short Stay Medical Unit is partially related to perfomance status, age, social and psychological status, admission type an the main diagnosis. PMID- 20011957 TI - [Diagnostic value of frozen section biopsy during surgery for breast lesions or neoplasms]. AB - BACKGROUND: During the surgical treatment of breast neoplasms (benign or malignant), frozen section biopsy is frequently requested to assess the kind of lesion and determine the surgical margins. AIM: To assess the diagnostic yield of frozen section breast biopsy. MATERIAL AND METHODS: AH the pathological reports of frozen section biopsies and definitive biopsies of 337 women aged 26 to 88 years, operated for suspected breast neoplasms between 2002 and 2006, were reviewed. The sensitivity, specificity and predictive value of frozen section biopsy, were calculated using the definitive biopsy as the gold standard. RESULTS: The definitive biopsy confirmed the presence of cancer in 290 women (86%). There were two false negative (0.59%) and no false positive frozen section biopsies for cancer detection. The sensibility for cancer detection was 99.3% and the specificity 100%. The positive predictive value was 100% and the negative predictive value 96.1%. The diagnosis of phyllodes tumor was missed by frozen section biopsy in three cases. The margins were informed in the 258 frozen section biopsies (79%) and in 59 cases (18%), these were positive for cancer. CONCLUSIONS: Frozen section biopsy is useful and reliable for cancer detection and margin status assessment in breast cancer surgery. PMID- 20011958 TI - [Cardiovascular risk profile and insulin resistance according body mass index, waist circumference and hypertriglyceridemic waist in adult subjects]. AB - BACKGROUND: Insulin plays a determinant role in the individual metabolic profile. Obesity, the most common cause of insulin resistance (IR), can be diagnosed by body mass index (BMI). Waist circumference (WC), specifically, measures accumulation of abdominal fat. The cardiovascular risk of an individual is strongly related to a combination of abdominal obesity with hypertriglyceridemia or hypertriglyceridemic waist (HTW). AIM: To assess the metabolic profile of cardiovascular risk and RI according to BMI, WC and HTW in adult patients. MATERIAL AND METHODS: Weight, height, BMI, blood pressure (BP), WC, HTW, blood glucose, total cholesterol, HDL cholesterol, LDL cholesterol, triglycerides and insulin were assessed and IR calculated by the homeostasis model assessment (HOMA) in 63 females aged 46+/-11 years and 12 males aged 4 7+/- 10 years. RESULTS: Differences in weight, WC, BP, lipids, glucose, insulin and HOMA, were observed as BMI increased. When patients were grouped according to WC, there were significant differences only in systolic pressure, triglycerides, insulin and HOMA. Individuals with HTW had a more adverse metabolic profile than those without HTW or with only high WC. A multiple regression equation determined that body mass index, total cholesterol and diastolic blood pressure were independent predictors of insulin resistance. CONCLUSIONS: Patients with HTW displayed an unfavorable cardiac risk profile. PMID- 20011959 TI - [Factors associated with maintenance of contraception among adolescents]. AB - BACKGROUND: Factors such as personal issues, family, sexuality and sexual partner characteristics are strongly associated with contraceptive continuation among single, nulliparous female adolescents. AIM: To determine factors associated to contraceptive maintenance among female nulliparous adolescents. MATERIAL AND METHODS: A cohort of 2,811 adolescents, who confidentially requested contraception in a sexual and reproductive health university center from 1990 to 2006 was analyzed. Two years after the request, their clinical records were reviewed to determine the time and length of contraception. Using life table analysis, the variables related to continuation or discontinuation of contraception were identified. RESULTS: Factors associated with a longer contraceptive use were a lower age at the moment of initiating the method, a better academic achievement and aspirations, higher schooling of the partner, higher age of the mother, having an adolescent mother, supervision of permissions by people different than parents and not attending to religious services. Variables associated with a higher risk for abandonment were a higher age of the adolescent, greater number of sexual partners, lack of communication with parents, non-catholic religious affiliation, use of oral hormonal contraceptive, greater number of siblings, commenting sexual issues with relatives or friends, having a partner without academic activity or working and to live without parents. CONCLUSIONS: Several personal, familial and environmental factors influence contraceptive use continuity among adolescents. PMID- 20011960 TI - [Non-toxigenic hemolytic Vibrio cholerae non-O1 non-O139 fatal septicemia. Report of one case]. AB - We report a 70-year-old woman, who had recently consumed shellfish, that was admitted to the intensive care unit with septic shock and died 19 hours later due to a multi-organic failure. Microbiological, serological and molecular assays confirmed a hemolytic tdh+ Vibrio cholerae non-01, non 0139 as the etiologic agent. PMID- 20011961 TI - [Gastrointestinal stromal tumor (GIST) in a patient with neurofibromatosis type 1. Report of one case]. AB - Neurofibromatosis Type 1 (NF1) is an autosomic dominant condition affecting the central nervous system and presenting a disposition towards development of gastrointestinal stromal tumors (GIST). We report a 38 year-old female patient with neurofibromatosis type 1 that required emergency surgery due to a perforated GIST originating in the fourth duodenal portion. The GIST, and the fourth duodenal portion, were excised and a primary duodenum-jejunal anastomosis was performed. The pathological study showed a partially necrotic solid-cystic tumor with 1 to 2 mitoses per 50-high-power fields. The cells stained positively to CD 117, CD34 and Desmin, and were negative to S-100, Vimentin, and Smooth Muscle a Actin. The patient is currently asymptomatic and under follow-up during the last 11 months after surgery. PMID- 20011962 TI - [Fatal neonatal sepsis caused by vertical transmission of Morganella morganii. Report of one case]. AB - We report a term neonate who developed early-onset sepsis due to Morganella morganii. The child was vaginally delivered after a short labor, and presented signs of perinatal asphyxia. Blood cultures taken soon after birth and from mother's lochia were positive for this microorganism. The infection was unresponsive to treatment with cefotaxime, to which the microorganism was susceptible, and the infant died at 17 days of age. M morganii is an opportunistic and uncommon pathogen, causing disease mainly in patients with underlying illness or after surgery. It is a rare perinatal pathogen, causing severe disease in premature infants, in association with maternal chorioamnionitis and premature rupture of the membranes. PMID- 20011963 TI - [Endemic pemphigus foliaceus in a pregnant woman. Report of one case]. AB - Endemic pemphigus foliaceus (EPF) in an autoimmune skin disease present in areas of the Amazonia. We report a 36 year-old woman who presented EPF at 17 weeks of pregnancy. At 29 weeks, she started antimicrobial treatment and steroids. At the moment of delivery, the disease was in remission and cutaneous lesions were not seen in the neonate. Indirect immunofluorescence titers of total IgG in the mother and in the neonate were negative. Sixteen months later, IgG titers in the offspring were 1/20 and remained negative in the mother, who was on low doses of oral corticosteroids. PMID- 20011964 TI - [Hypocretin system and narcolepsy]. AB - Narcolepsy is a sleep disorder characterized by excessive daytime sleepiness, cataplexy, hypnagogic hallucinations, and sleep-onset rapid eye movement (REM) sleep periods. It is now identified as a neurodegenerative disease because there is a massive loss of specific neurons in the brain. These neurons contain the neuropeptides hypocretin-1 and hypocretin-2, which are also known as orexin-A and orexin-B. Cerebrospinal fluid hypocretin-1 measurements are diagnostic for primary narcolepsy. The cause of neural loss could be autoimmune since most patients have the HLA DQB1 0602 allele that predisposes to the disorders. The discovery of hypocretin deficiency is redefining the clinical entity of narcolepsy and offering novel diagnostic procedures. This article reviews the current understanding of narcolepsy and discusses the implications of hypocretin discovery. PMID- 20011965 TI - [Association between air pollution and cardiovascular risk]. AB - A clear cut relationship between particulate matter air contamination and the mortality and morbidity due to respiratory disease has been observed in the last decades. However there is also a relationship between air pollution and cardiovascular diseases. In big cities, a big or small particle concentration increase of 10 micro/m(3) is associated with a significantly higher risk of ischemic heart disease and myocardial infarction, both when acute or chronic exposures are considered. The risk is higher for small particles. Similar risk increases are observed in patients with hypertension, stroke or severe arrhythmias. This association is independent of environmental distracters such as weather, temperature or humidity and of classical cardiovascular risk factors such as age, diabetes, dyslipidemia and obesity. Physicians should be aware of the problem and explain their patients the increased risk that they are facing due to air pollution. PMID- 20011966 TI - [Leptin-melanocortin system, body weight regulation and obesity]. AB - Obesity is a multifactorial disease that is rarely associated to single gene defects. However, due to their direct cause-effect relationships, those genetic defects that cause some forms of monogenic obesity are relevant in the study of mechanisms that contribute to increased energy intake and body fat accumulation. Most of the genes that have been shown to cause monogenic obesity are related to the leptin-melanocortin system. The functionality of this system has been elucidated through natural mutations (Agouti, ob and db) in mice and knock-out models. Mutations related to human monogenic obesity have been described in leptin, leptin receptor, proopiomelanocortin, prohormone convertase 1 or melanocortin receptor 4 genes. Therapy with human recombinant leptin in patients with genetic deficiency of the hormone is an effective medical treatment of obesity, although only applicable to very few families. The use of leptin melanocortin agonists, drugs to avoid leptin resistance or combinations of treatments with leptin and other satiating peptides are currently being investigated for multifacotiral human obesity. PMID- 20011967 TI - [Ignacio Matte Blanco, MD, and the development of psychiatry teaching to medical students]. AB - Ignacio Matte Blanco had an important role in organizing the teaching of Psychiatry in medical school. In this paper we describe his training, that began as a physiologist, but turned into psychiatric and psychoanalysis in his formation in the United States and the United Kingdom during the forties. After returning to Chile, in the Chair of Psychiatry at the Faculty of Medicine of the Universidad de Chile he developed pioneering ideas about undergraduate teaching of psychiatry, that were exposed not only in Chile but to the Pan-American Health Organization. He advocated decreasing the time spent in lectures, and increase clinical practice and group dynamic experiences centered in the students. He insisted that teaching had to be focused in issues useful for general physicians and non psychiatric specialists, as well as in the need to extend the psychosocial curriculum to the internship. He also pointed to the need of increasing the humanistic formation of medical students. When Matte Blanco emigrated to Rome in 1966, his influences seemed to wane, since most of his disciples inverted exclamation markeft the Clinica Psiquiatrica Universitaria, where he taught. However, since the eighties, several of his viewpoints have been included in the medical curricula of Chilean schools of medicine. PMID- 20011968 TI - [Update of consensus recommendations of the Chilean Hypertension Society about ambulatory blood pressure monitoring]. AB - Ambulatory blood pressure monitoring (ABPM) is a valuable tool to evaluate the blood pressure pattern, to identify hypertensive patients, to diagnose white coat and masked hypertension and in situations in which a tight control of hypertension is crucial. This is an update of 1999 consensus recommendations about the use to ABPM, considering that there is new evidence concerning its benefits, and the clinical experience with its application has increased. Equipment programming, its installation, the interpretation and analysis of the data are described, and a report sheet for patients is included. New recommendations have been added to the accepted indications. Normal blood pressure ranges for children and pregnant women have been replaced by new data. PMID- 20011969 TI - [Sixty years of mitral valve surgery]. AB - On June 10, 1948, Charles Bailey, MD, operated successfully a mitral valve stenosis in Philadelphia and six days later, Dwight Harken, MD, performed the same operation in Boston, marking the onset of cardiovascular surgery. These successful operations were preceded by several failures and even deaths, that had to be overcome by both pioneers. This manuscript reviews several cases and situations that these surgeons had to face during the development of cardiovascular surgery, that changed the natural history of cardiac diseases. The history culminates with the successful mitral valve replacement, performed by Albert Starr using a mechanical valve designed by him and Lowell Edwards, a retired engineer whose primary interest was to develop an artificial heart. The first mitral commissurotomy in Chile was performed by Svante Tornvall, MD, and Pedro Uribe, MD, at van Buren Hospital in Valparaiso in December, 1950. The first mitral valve replacement was performed by Hugo Salvestrini, MD, in 1964 at the Catholic University Hospital. PMID- 20011970 TI - [Are blood glucose measurements made with portable glucometers reliable?]. AB - Examinations performed beside the bed of patients ("Point-of-care testing, POCT") provide immediate results and are simple to perform. The most common of these tests is the self control of blood glucose levels in diabetic patients. The use of these devices at the hospital level, introduces a new set of requirements to health institutions, which should monitor all aspects of the process, including training of final users, proper quality control, development of written procedures for use and even participation in surveys of external quality control, avoiding the generation of errors and guaranteeing patient safety. PMID- 20011971 TI - [Scientific journals for medical students in Chile]. PMID- 20011972 TI - Discovery of YB-1 as a new immunological target in neuroblastoma by vaccination in the context of regulatory T cell blockade. AB - Neuroblastoma is one of the most common solid tumors in infancy and early childhood. Using the A/J mouse and a syngeneic neuroblastoma cell line AGN2a, we induced a strong anti-neuroblastoma cellular immune response when AGN2a transfected to express costimulatory molecules (CD80/CD86/CD54/CD137L) was used as a vaccine in the context of regulatory T cell blockade. Strong humoral immunity was induced by AGN2a-4p immunization in the context with regulatory T cell blockade. Serum from treated mice was used to screen an AGN2a cDNA expression library that was constructed with lambda ZAP express vector in order to identify tumor-associated antigens by SEREX. Twenty one clones were identified by sequencing and comparative analysis of gene pools. Most transcripts play some roles in the neuronal differentiation, cell metabolism, or have previously been identified as transcripts that are over-expressed in other malignancies. The most commonly identified tumor-associated antigen, using serum from AGN2a-4p immunization with Treg blockade mice, was YB-1 protein that also induced a T cell response. These results indicated that potential neuroblastoma-associated antigens were found by the sera from mice immunized with tumor cells expressing costimulatory molecules with regulatory T cell function blockade. The identification of YB-1 as tumor-associated antigens capable of eliciting a T cell response validates our experimental approach and argues for the antigens we have identified here to be evaluated as targets of effector immunity and as vaccine candidates. PMID- 20011973 TI - B23 interacts with PES1 and is involved in nucleolar localization of PES1. AB - PES1, the human homolog of zebrafish pescadillo, is a nucleolar protein that is essential for cell proliferation. We report herein that a nucleolar marker protein B23 physically interacts with PES1 and is involved in the nucleolar localization of PES1. In vivo interaction between B23 and PES1 was verified by co immunoprecipitation of endogenous B23 and PES1 proteins, and they showed cellular co-localizations under both normal and actinomycin D-induced stress conditions. Furthermore, we mapped their interaction domains via in vitro pulldown assays. When B23 was knocked down by RNA interference, there appeared an increased nucleoplasmic distribution of PES1. Our results support a previous hypothesis that B23 might be a nucleolar hub protein for protein targeting to the nucleolus, and shed light on the nucleolar localization mechanism of PES1. The physical interaction between B23 and PES1 implies that they may participate in ribosome biogenesis in a protein complex. PMID- 20011974 TI - Recovery of infectious foot-and-mouth disease virus from full-length genomic cDNA clones using an RNA polymerase I system. AB - The prototypic foot-and-mouth disease virus (FMDV) was shown more than a century ago to be the first filterable agent capable of causing FMD, and it has served as an important model for studying basic principles of Aphthovirus molecular biology. However, the complex structure and antigenic diversity of FMDV have posed a major obstacle to the attempts at manipulating the infectious virus by reverse genetic techniques. Here, we report the recovery of infectious FMDV from cDNAs based on an efficient in vivo RNA polymerase I (polI) transcription system. Intracellular transcription of the full-length viral genome from polI-based vectors resulted in efficient formation of infectious virus displaying a genetic marker. Compared with wild-type virus, an abundance of genomic mRNA and elevated expression levels of viral antigens were indicative of the hyperfunction throughout the life-cycle of this cDNA-derived virus at transcription, replication, and translation levels. The technology described here could be an extremely valuable molecular biology tool for studying FMDV complex infectious characteristics. It is an operating platform for studying FMDV functional genomics, molecular mechanism of pathogenicity and variation, and lays a solid foundation for the development of viral chimeras toward the prospect of a genetically engineered vaccine. PMID- 20011975 TI - Genome-wide identification of target genes repressed by the zinc finger transcription factor REST/NRSF in the HEK 293 cell line. AB - Transcriptional repression is as important as transcriptional activation in establishing cell-type specific patterns of gene expression. RE1-silencing transcription factor (REST), also known as neuronal restrictive silencing factor (NRSF), is a transcriptional regulator that represses a battery of neuronal differentiation genes in non-neuronal cells or in neural progenitor cells by binding to a specific DNA sequence (repressor element-1/neuron-restrictive silencer element, RE1/NRSE). REST/NRSF functions in the neuronal development are widely studied, however, little is known about target genes in various non neuronal lineages that may result in cell differentiation. Here, we use RNA interference (RNAi) technology combined with the microarray strategy to identify potential REST/NRSF targets and RE1/NRSEs in human non-neuronal cell line HEK 293. Expression of 54 genes was up-regulated by inhibition of REST/NRSF in the HEK 293 cells according to the microarray experiment and 13 of those were further confirmed by quantitative RT-PCR. Our results confirmed the good confidence and reliability of current research data based on in silico, chromatin immunoprecipitation in combination with microarrays (ChIP-chip), and high throughput sequencing (ChIP-seq). However, in view of the fact that thousands of genes have been testified or predicted to be recognized by REST/NRSF, our data show that only a few genes among those are directly up-regulated by the interaction of REST/NRSF with RE1/NRSEs sites in gene sequences. PMID- 20011976 TI - Potentiation of (-)-epigallocatechin-3-gallate-induced apoptosis by bortezomib in multiple myeloma cells. AB - The green tea constituent, (-)-epigallocatechin-3-gallate (EGCG), has chemopreventive and anticancer effects. This is partially because of the selective ability of EGCG to induce apoptosis and death in cancer cells without affecting normal cells. In the present study, the activity of EGCG against the myeloma cell line, KM3, was examined. Our results demonstrated, for the first time, that the treatment of the KM3 cell line with EGCG inhibits cell proliferation and induces apoptosis, and there is a synergistic effect when EGCG and bortezomib are combined. Further experiments showed that this effect involves the NF-kappaB pathway. EGCG inhibits the expression of the P65 mRNA and P65/pP65 protein, meanwhile it downregulates pIkappaBalpha expression and upregulates IkappaBalpha expression. EGCG also activates caspase-3, -8, cleaved caspase-9, and poly-ADP-ribose polymerase (PARP) and subsequent apoptosis. These findings provided experimental evidence for efficacy of EGCG alone or in combination with bortezomib in multiple myeloma therapy. PMID- 20011977 TI - Hyperhomocysteinemia stimulates hepatic glucose output and PEPCK expression. AB - Homocysteine is an intermediate in the sulfur amino acid metabolism. Recent studies suggested that there might be links between hyperhomocysteinemia and insulin resistance. In the present study, we investigated the effect of homocysteine on glucose metabolism. We demonstrated that the levels of insulin were significantly higher in mice with hyperhomocysteinemia than those in the normal mice after administration of glucose. The effect of insulin on glucose output was significantly blocked in the homocysteine-treated hepatocytes. In addition, the expression of phosphoenolpyruvate carboxykinase (PEPCK) gene was elevated in the liver of mice with hyperhomocysteinemia and primary mouse hepatocytes treated with homocysteine. The action of homocysteine was suppressed by H89, a protein kinase A (PKA) inhibitor. Thus, hyperhomocysteinemia may be considered as a risk factor that contributes to the development of insulin resistance with respect to elevated glucose output and upregulation of PEPCK, probably via the PKA pathway. Our study provides a novel mechanistic explanation for the development of insulin resistance in hyperhomocysteinemia. PMID- 20011978 TI - Antioxidant activity of mangostin in cell-free system and its effect on K562 leukemia cell line in photodynamic therapy. AB - Mangostin (MAG), a kind of xanthone widely used in diet and medicine, has antioxidant, anti-inflammatory, antimicrobial, and anticancer activities. On account of its antioxidant activity, MAG might protect cancer cells from free radical damage in photodynamic therapy (PDT) during which reactive oxygen species production was stimulated leading to irreversible tumor cell injury. In this study, the antioxidant activity of MAG was investigated and the influence of MAG on K562 cells in 5-aminolevulinic acid (ALA)-based PDT is demonstrated. The results showed that MAG could scavenge hydroxyl radical, superoxide anion, and hydrogen peroxide and inhibit the formation of malondialdehyde (MDA), but increase the amounts of singlet oxygen in cell-free systems. MAG inhibits cell proliferation and enhances cell apoptosis, lipid peroxidation, and DNA damage in ALA-PDT on K562 cells. NaN3, a singlet oxygen quencher, suppresses the MAG induced cell apoptosis, lipid peroxidation, and DNA damage. In conclusion, MAG enhances the PDT-induced cytotoxicity in K562 cells and singlet oxygen was involved in this process. These results implied that the effect of antioxidants on PDT might be determined by its sensitization ability to singlet oxygen. PMID- 20011979 TI - Modeling of protein refolding from inclusion bodies. AB - Overexpression of foreign proteins in Escherichia coli often leads to the formation of inclusion bodies (IBs), which becomes the major bottleneck in the preparation of recombinant proteins and their applications. In the present study, 36 proteins from IBs were refolded using a simple refolding method. Refolding yields of these proteins were defined as the percentage of soluble proteins following dilution refolding in the amount of denatured proteins in the samples before diluting into refolding buffer. Furthermore, a mathematical model was deduced to evaluate the role of biochemical properties in the protein refolding. Our results indicated that under the experimental conditions, isoelectric point of proteins might be mostly contributing to the high efficacy of protein refolding since the increment of one unit resulted in a decrease of 14.83% in the refolding yield. Other important mediators were components of protein secondary structure and the molecular weight (R(2) = 0.98, P = 0.000, F-test). Six proteins with low efficiency in the protein refolding possessed relatively low isoelectric points. Furthermore, refolding yields of six additional proteins from IBs were predicted and further validated by refolding the proteins under the same conditions. Therefore, the model of protein refolding developed here could be used to predict the refolding yields of proteins from IBs through a simple method. Our study will be suggestive to optimize the methods for protein refolding from IBs according to their intrinsic properties. PMID- 20011980 TI - A non-viral vector for potential DMD gene therapy study by targeting a minidystrophin-GFP fusion gene into the hrDNA locus. AB - Gene therapy has emerged as a promising approach for the lethal disorder of Duchenne muscular dystrophy (DMD). Using a novel non-viral delivery system, the human ribosomal DNA (hrDNA) targeting vector, we targeted a minidystrophin-GFP fusion gene into the hrDNA locus of HT1080 cells with a high site-specific integrated efficiency of 10(-5), in which the transgene could express efficiently and continuously. The minidystrophin-GFP fusion protein was easily found to localize on the plasma membrane of HT1080 cells, indicating its possible physiologic performance. Our findings showed that the hrDNA-targeting vector might be highly useful for DMD gene therapy study. PMID- 20011981 TI - Proteolytic regulatory mechanism of chemerin bioactivity. AB - Chemerin is a novel chemoattractant recognized by chemokine-like receptor 1 (CMKLR1), a serpentine receptor expressed primarily by plasmacytoid dendritic cells, natural killer cells, and macrophages. Human prochemerin circulates in plasma as an inactive precursor. Its chemotactic activity is expressed upon cleavage of the C-terminal amino acid residues by proteases of the coagulation, fibrinolytic, and inflammatory system. The C-terminal cleavage site of prochemerin is highly conservative, indicating that the proteolytic regulation of chemerin bioactivity is a common mechanism undertaken by different species. In this review, we summarized chemerin-proteases interactions, chemerin receptors, and their importance in normal and pathologic conditions. PMID- 20011982 TI - Work-related and psychological determinants of multisite musculoskeletal pain. AB - OBJECTIVE: Musculoskeletal pain is associated with occupational physical activities and psychosocial risk factors. We evaluated the relative importance of work-related and psychological determinants of the number of -anatomical sites affected by musculoskeletal pain in a cross-sectional survey. METHODS: The survey focused on musculoskeletal pain in six body regions (low-back, neck, shoulder, elbow, wrist-hand, and knee) among 224 nurses, 200 office workers and 140 postal clerks in Crete, Greece (response rate 95%). Information was collected about demographic characteristics, occupational physical load, psycho-social aspects of work, perceptions about the causes of pain, mental health, somatization, and experience of pain in the past 12 months. We used Poisson regression to assess associations of risk factors with the number of painful anatomical sites and explored interactions using classification and regression trees (CART). RESULTS: Two-thirds of the study sample reported pain in >or=2 body sites during the past 12 months, and in 23%, >3 sites were affected. The number of painful anatomical sites was strongly related to both physical load at work and somatization (with relative risks increased 5-fold or more for frequent and disabling multisite pain) and was also significantly associated with work-related psychosocial factors and beliefs about work causation. The CART analysis suggested that somatization was the leading determinant of the number of painful body sites. CONCLUSION: In the population studied, pain at multiple anatomical sites was common and strongly associated with somatization, which may have a more important influence on multisite pain than pain that is limited to a single anatomical site. PMID- 20011983 TI - Pregnancy outcomes among daycare employees in Finland. AB - OBJECTIVE: The aim of this study was to investigate whether working as a daycare employee increases the risk of perinatal death, pre-term birth, low birth weight, smallness for gestational age, or congenital malformations. METHODS: We conducted a register-based cohort study among daycare employees and women from various occupations of healthcare (reference group). Study subjects were identified from the files of Finnish trade unions and the National Authority for Medicolegal Affairs. Pregnancy outcomes, antenatal occupation, and working status were obtained by linkage to national registers. The final data consisted of 13 299 and 12 182 singleton births in the study and reference groups, respectively. We analyzed pregnancy outcome data using generalized estimating equations and linear regression. RESULTS: The occurrences of pre-term birth [odds ratio (OR) 0.91, 95% confidence interval (95% CI) 0.79-1.06], perinatal death (OR 0.91, 95% CI 0.62 1.34), smallness for gestational age (OR 1.01, 95% CI 0.91-1.12), and congenital malformation (OR 1.10, 95% CI 0.92-1.32) were similar among the children of the daycare employees and the reference group. The adjusted mean birth weight of the children of the daycare employees was slightly higher (14 g, 95% CI -1-29) than that of the reference group, but the difference was attenuated to 6 g in the subset of the first births. CONCLUSION: Daycare employees were not, in general, at an increased risk of an adverse pregnancy outcome. However, efforts should nevertheless be made to prevent their exposure to harmful viruses and heavy physical load during pregnancy. PMID- 20011984 TI - Measures to counteract the negative effects of night work. AB - OBJECTIVES: Night work is associated with several negative health outcomes as well as accidents and reduced productivity. The aim of this study was to identify factors that may counteract the negative effects of night work. METHODS: We conducted searches for studies of scientifically based countermeasures of the negative effects of night work in PubMed and Thomson Reuters' ISI Web of Knowledge and inspected the reference lists of relevant literature. RESULTS: We identified studies describing countermeasures such as proper personnel selection, bright light therapy, melatonin administration, naps, exercise, sleepiness detection devices, and the use of stimulants to improve wakefulness and hypnotics to improve daytime sleep. CONCLUSIONS: There is some research supporting countermeasures (eg, bright light, melatonin, naps, use of stimulants, proper work scheduling) as a means to improve adaptation to night work. However, there is little evidence that such countermeasures reduce the long-term health consequences of night work. Future studies should aim at identifying both work and individual factors which are related to differential health outcomes of night work. Better study designs (eg, longitudinal designs and use of standardized outcome measures) are needed in future research. PMID- 20011985 TI - Club health: global perspectives, new directions. AB - This editorial presents the views of the guest editors of this collection in summarising some of the general themes arising from a selection of papers presented at the 2008 Club Health Conference in Ibiza, Spain. Particular emphasis is placed upon the value of comparative approaches to club studies, youth culture and substance use so that researchers, policy-makers, and practitioners can gain new understanding of the interplay of local and global factors which shape club cultures, nightlife scenes, and individual lifestyles. By contextualizing the studies in this volume, the editorial outlines important themes and issues which will help promote Club Studies as an interdisciplinary field of endeavour well into the future. PMID- 20011986 TI - Alcohol, drugs, sex and violence: health risks and consequences in young British holidaymakers to the Balearics. AB - BACKGROUND: Substance use, risky sexual behaviour and violence are among the key youth health issues today. Whilst they are the focus of much prevention work in the UK, relatively little information is available to inform prevention in international holiday resorts, where young people can take the greatest risks with their health. METHODS: We conducted a cross-sectional survey of 1033 British holidaymakers aged 16-35 in Ibiza and Majorca airports (Spain). Surveys covered frequency of alcohol and drug use at home and abroad, and drunkenness, sexual activity and experience of a range of negative experiences on holiday. RESULTS: Illicit drug use was more common in Ibiza. Over half (53.9%) of visitors to Ibiza reported using at least one drug on holiday (compared with 13.9% in Majorca) and over a third reported using more than one drug. However visitors to Majorca reported more frequent drunkenness and more negative outcomes (arguments, fighting, unintentional injury, illness and requiring medical attention). In both locations, frequency of substance use increased on holiday. A quarter of those travelling without a partner reported having had sex during their holiday, and of these a third did not consistently use condoms. Frequent drunkenness was associated with fighting, unintentional injury and sex with a new partner. Illicit drug use, particularly polydrug use, was associated with fighting and unprotected sex. CONCLUSIONS: Young holidaymakers face significant health risks on holiday. Understanding which resorts are associated with which types of health risk behaviour is critical in targeting health promotion and harm reduction measures appropriately. PMID- 20011987 TI - Evaluation of a designated driver intervention to prevent alcohol-related road accidents in the clubs of Milan, Italy. AB - OBJECTIVE: The present study is an evaluation of a designated driver intervention. The objectives were: (1) to verify the intervention assumption, which is that offering incentives, linked to a positive result of the breathalyser test (<0.5 g/l) when leaving the club, leads to a decrease in the percentage of drivers leaving the club with a BAC above the legal limit; (2) to understand the reason why drivers take part in the intervention and assess whether the operation was effective in lowering the amount of alcohol consumed during the night; (3) to verify the assumption that there is a bias in the selection of designated drivers. METHOD: Pre-experimental research design with a control group and no pre-test. The two conditions of the independent variable were: presence or absence of the "Safe Driver" intervention. A group of drivers who took part in the intervention (DDs N=124) was compared with a group that did not (non-DDs N=139). RESULTS: (1) DDs' mean BAC was significantly lower than non DDs', but (2) the incentive motivated just 5% of DDs to drink less, and (3) there is a bias in the selection of DDs that present a lower drinking risk profile than non-DDs. DISCUSSION: The effectiveness of the intervention doesn't seem to be supported by strong evidence. The free-entrance incentive is not effective in motivating drivers to drink less. More research is needed to evaluate the effectiveness of designated driver interventions where the bias in the selection of drivers has been removed, by, for example, using other incentives and to have a clear understanding of designated drivers' risk profile. PMID- 20011988 TI - Variations in clubbers' substance use by individual and scene-level factors. AB - Research on clubbing spans numerous fields and adopts diverse approaches to the populations in question. Yet, a somewhat homogenous narrative has emerged about clubbers and substance use, one that focuses on overconsumption, risky behavior, and considerable consequence. The purpose of our study is to unpack this narrative by exploring how the substance use patterns of clubbing populations differ by individual and scene-level factors. From this, we hope to increase an understanding of how social and cultural factors impact the relationship between clubbers and substance use. We draw on direct observation of 29 club events and interviews with 51 respondents from the hip hop (HH) and electronic dance music (EDM) scenes in Philadelphia to inform our research objective. Analyses revealed two broad criteria on which a substance use-based typology of clubbers can be offered: motivations for clubbing and level and type of scene involvement. From these two dimensions, we are able to distinguish among three types of clubbers: drug sub-cultural members, commercial clubbers, and music connoisseurs. Drug sub cultural members reported early onset of drug use and a greater frequency of current drug use at largely underground EDM club events and outside them as well. Commercial clubbers had benign histories with drugs, but they reported attending mostly commercialized HH events to get drunk and court the opposite sex. Music connoisseurs reported minimal current drug and alcohol use at exclusively underground EDM and HH club events, following from, in some cases, extensive histories with drugs and alcohol. The differences between these three groups' substance use patterns, we contend, can be attributed to the interplay between numerous social and cultural factors not previously considered. PMID- 20011989 TI - Drugs and alcohol consumption and sexual behaviours in night recreational settings in Portugal. AB - BACKGROUND: Night recreational settings have become a key location for the socialisation of young people. For customers attending these places, the consumption of alcohol and drugs is related to changes in sexual decision making and associated with increased likelihood of engagement in more risky sexual behaviours. AIM: To identify the characteristics of Portuguese young people who attend nightclubs, pubs and bars; to explore relationships between attendance at these settings and the consumption of alcohol and drugs; to study associations between substance use and sexual behaviour. METHODS: Respondent-driven sampling methodology was used in nine Portuguese cities to survey 1142 young people who attend nightlife leisure locations. RESULTS: Respondents regularly go out at night, especially at weekends. The most used substances are alcohol, cannabis, cocaine and ecstasy. Alcohol and drugs users have more regular and deeply-rooted night recreation habits than nonusers and drugs consumption increases with increased recreational activities. In the last 12 months, 83.3% of the respondents have already had sexual intercourse (X= 2.0 partner; SD = 2.3), 51.0% had sex under the influence of alcohol, 22.9% under the influence of drugs and 46.7% agreed that this influence made them have unsafe sex. Early alcohol and drug use was proportionately related to an early sexual experience. Alcohol and drugs use s were more likely to adopt more sexual risk behaviours than non-users. CONCLUSIONS: The enjoyment of recreational nightlife settings exposes Portuguese clubbers to environmental factors which, in combination with substance use, may influence the nature of relationships between alcohol and drug use and sexual behaviour. PMID- 20011990 TI - 'Lager, lager shouting': the role of music and DJs in nightclub disorder control. AB - OBJECTIVE: This paper explores how music and music genre can govern the nightlife experience, specifically how a nightclub's music policy can impact on clientele, health behaviours, bar sales and levels of disorder. MATERIAL AND METHOD: Participant observations were conducted in sample of eight city centre nightclubs in Glasgow, Scotland. RESULTS: Music policy was found to influence nightclubs' clientele and their behaviours, for example in relation to differences in levels of alcohol or illegal drug use, sexual activity and violence between venues. Further, within individual venues, music policy was also observed operating as a crowd control tool, with for example such entertainment being used in alcohol marketing, in 'soft policing' and in the temporal management of patrons' movements. This research acknowledges the centrality of the DJ in implementing these controls. CONCLUSIONS: It is clear that music is a central component of the Nighttime Economy. Despite this, to date music has tended to play 'second fiddle' to sex and drugs within Club Health research. From this research it is recommended that music policy be incorporated into responsible bar server training packages. PMID- 20011991 TI - Patterns and harms of pharmaceutical drug use among ecstasy users in Australia. AB - BACKGROUND: There has been a significant incursion of prescription drugs in to the club scene worldwide. The concomitant use of ecstasy and pharmaceutical drugs is associated with potentially serious health-related harms. This has ramifications for people who deliberately combine pharmaceutical drugs with ecstasy to achieve a specific effect and for people legitimately using pharmaceutical drugs for a health condition who may incidentally use ecstasy. This exploratory study had the broad aim to investigate the patterns and harms of concomitant use of pharmaceutical drugs and ecstasy, and to explore participants' experiences when visiting a General Practitioner (GP). METHODS: In-depth interviews were conducted with 30 ecstasy/pharmaceutical drug users. RESULTS: Users reported deliberately combining stimulants (dexamphetamine, methylphenidate (e.g. Ritalin), antidepressants and sildenafil (e.g. Viagra) for non-medical purposes with ecstasy. A wide range of effects were reported. Several clear relationships emerged between the use of pharmaceutical drugs and ecstasy, and the distinctions between these in terms of motivations for use and consequences are discussed. The attitudes of GPs were professional and non-judgemental and there was little reluctance among participants to disclose illicit drug use. Screening for illicit drug use was limited. IMPLICATIONS: Findings have important implications for clubbers in regard to harm reduction strategies and highlight a number of areas for future research. PMID- 20011992 TI - Everything starts with an 'E': exclusion, ethnicity and elite formation in contemporary English clubland. AB - Early Club Studies emphasised the inclusiveness of club cultures and the PLUR ethos of 'peace, love, unity and respect' alongside a polarised characterisation of nightlife contexts, as either commercial, alcohol-oriented nightclubs offering 'mainstream' pop music, or 'authentic'/'alternative' underground dance clubs associated with widespread illicit drug use. This paper adds to the growing body of research problematising these simplistic characterisations of club cultures and leisure venues across the night-time economy, emphasising elements of fragmentation and segregation alongside the continued importance of social structure and resultant social and spatial exclusion. The authors explore how informal processes - such as club launches, internet promotions and dress codes - together result in the production and reproduction of two contrasting forms of English clubland elites: 'cultural elites' produced through the social, cultural and spatial exclusion of electronic dance music of black origin and its minority ethnic, working class and lower income followers from Manchester city centre dance clubs; and 'consumer elites' produced through the economic and cultural exclusion of working class and lower income club-goers from nightclubs in London's West End. The complex and interweaving practices of cultural distinction and structural discrimination which produce such elites are often closely intertwined with the formal and informal regulation, marginalization and criminalization of specific cultural forms. The paper therefore argues for the construction of more nuanced conceptual understandings of the social divisions and inequalities within nightlife and in studies of young people's leisure opportunities more generally. PMID- 20011993 TI - Preventive interventions in nightlife: a review. AB - Participating in nightlife it is a meaningful cultural activity for young people. Councils welcome a flourishing late-night entertainment industry. But there also problems related to the people involved (alcohol and drug abuse, violence, risky sex...) and to the recreational activity itself (noise, street drinking, vandalism...). Prevention intervention possibilities are numerous, and most have been implemented, however assessment of their outcomes is less common. We have identified 11 types of prevention according to their main objective (venues centred, community, education, law enforcement, industry collaboration...) and we have reviewed the existing evidence in relation to each form of intervention. Popular types of interventions as providing information or pill testing are not evidence based. Other interventions as Responsible beverage services or designated driver programmes, backed by the industry, are not exactly the most effective, especially if they are not enforced. Others like community approaches can be effective but it can be a problem how to achieve their continuity. From the present review, what emerges as the best strategy is the combination of training, cooperation and enforcement. 'Classical' measures (taxation, reduced BAC limits, minimum legal purchasing age...) are also evidence-based and effective. Could be interesting to investigate interventions that address the cultural aspects of nightlife. But this sort of interventions do not depend on the will or the skills of professionals, and are more related to societal changes. The highly frequent use of popular but ineffective interventions contributes more to the problem than to its solution, and only facilitates giving preventive efforts a bad reputation. PMID- 20011994 TI - [Clinical trials with advanced therapy medicinal products]. AB - For advanced therapies, the same basic principles for assessment apply as for any other biotechnological medicinal product. Nevertheless, the extent of data for quality, safety, and efficacy can be highly specific. Until recently, advanced therapies were not uniformly regulated across Europe, e.g., tissue engineered products were regulated either as medicinal products or medical devices. Thus, for some products no data from clinical studies are available, e.g., for autologous chondrocyte products. The draft guideline on Good Clinical Practice for clinical trials with advanced therapies describes specific additional requirements, e.g., ensuring traceability. Most clinical studies with advanced therapies in Germany are still in early phase I or II trials with highly divergent types of products and clinical indications. The Committee for Advanced Therapies (CAT) at the European Medicines Agency (EMEA) has been established to meet the scientific and regulatory challenges with advanced therapies. PMID- 20011995 TI - [Further training in surgery from the perspective of the German Society for Surgery]. PMID- 20011996 TI - [Surgical management of thyroid diseases]. AB - Operations of the thyroid gland belong to the most frequent surgical procedures in Germany. Ultrasound is used for examination of the morphology and scintigraphy for examination of the function of the thyroid gland. Cytology can be used to examine scintigraphically cold nodules. Nodules represent the most frequent indication for thyroid surgery. In differentiated thyroid carcinoma postoperative radioactive iodine therapy plays an important role. Operations of the thyroid gland should be performed with optical magnification to identify the recurrent laryngeal nerve with the help of neuromonitoring. A missing electric signal after stimulation of the nerve with intact morphology is indicative of temporary paresis of the vocal cord and operation of the contralateral side should be postponed. Thus, a bilateral paresis can be safely avoided. PMID- 20011997 TI - [Sphenoorbital meningiomas: results in long-term treatment]. AB - BACKGROUND: The purpose of this study was to present the findings on growth patterns in a large surgical series of sphenoorbital meningiomas. PATIENTS AND METHODS: A retrospective analysis was performed on 77 patients (61 f) harboring sphenoorbital meningiomas, who underwent surgery between 1991 and 2009. The standard surgical approach consisted of pterional craniotomy and extradural resection of any infiltrated bone. The intradural and orbital tumor was resected, with complete reconstruction of the dura while the lateral bone was partially reconstructed. The follow-up period ranged from 6 to 130 months (mean: 57.9 months). RESULTS: Total macroscopic resection was achieved in 42 patients. Fifty eight meningiomas extended to the orbital roof and/or lateral orbital wall, 42 involved the extraconal space, and 11 the intraconal space. Sixty-three tumors involved the superior orbital fissure, 54 the optic canal, and 24 the inferior orbital fissure. Seventeen tumors infiltrated the cavernous sinus and 37 involved the anterior clinoid process. The rate of minor morbidity was 14.3% (slight deficits) and the rate of major morbidity was 4% (significant deficits). Subtotal resections were performed on 35 patients because there was intraorbital tumor (n=8); tumor in the cavernous sinus (n=12); tumor invading the superior orbital fissure (n=12); and tumor of the skull base (n=3). Nine patients underwent postoperative three-dimensional conformal radiotherapy, which resulted in stable tumor volume at follow-up in eight patients. Tumor recurrence was identified in ten patients (12.9%) postoperatively (range of follow-up: 10-47 months). CONCLUSION: The goal of surgery is complete tumor removal without morbidity. Exact analysis of tumor growth and possible involvement of pertinent structures are mandatory in planning the procedure. PMID- 20011998 TI - [Indolent swelling of the neck]. AB - Scleredema adultorum Buschke is a rare disease of unknown etiology characterized by an immovable edematous swelling and hardening of the skin which typically begins in the neck and spreads to the face, head, shoulders and thorax. No standardized therapy for scleredema adultorum Buschke has yet been established. As in the case of our patient, a systemic cortisone therapy is normally administered combined with high-dose penicillin and physiotherapy. PMID- 20011999 TI - [Specific immunotherapy with depigmented allergoids]. AB - Specific immunotherapy is the only available causative treatment for IgE-mediated allergic conditions. The state of the art is treatment via the subcutaneous route with crude extracts in a water solution, with physically linked (semidepot) extracts or chemically modified semidepot extracts (allergoids). A relatively new purification method combines depigmentation followed by polymerization with glutaraldehyde. This modification results in increased tolerance with a reduction in both local and systemic adverse effects. As controlled clinical trials have shown, the effectiveness is comparable to that of specific immunotherapy with crude allergen extracts. Recent data suggest that the modified polymerized allergoids allow a safe rush titration in a few days or even in 1 day (ultra-rush titration). PMID- 20012000 TI - [Principles of radiation therapy for malignant tumors of the orbit and the periorbital area]. AB - Primary malignant tumors of the orbit and the orbital adnexal area are rare, variform and in the majority of cases need an interdisciplinary treatment. According to tumor entity, tumor localization, and tumor extent the complete spectrum of radiotherapeutic therapy options is necessary. In the majority of malignant tumors, such as those of the eyelids, lachrymal glands, orbit, metastases, sarcoma or lymphoma, radiotherapy is used as a high energy therapy by means of medical linear accelerators as so-called teletherapy. In addition, new therapy methods, such as stereotactic highly conformal radiation or intensity modulated radiation therapy (IMRT) were developed to achieve a higher local tumor control by a better configuration of the radiotherapy fields to the tumor borders and to contribute to a reduction of acute and chronic side effects in normal tissue by blurring of the dose. The application of ionizing radiation sources, such as so-called interstitial brachytherapy can also be just as carefully as effectively used. Particle beam radiotherapy is limited only to specially licensed centers and light hydrogen ions as proton therapy or carbon ions as heavy ion radiotherapy are in use. This overview will show the options as well as important progress in the field of radiotherapy in the treatment of tumors of the orbit and periorbital area. PMID- 20012005 TI - [Functional magnetic resonance imaging with ultra-high fields]. AB - Functional magnetic resonance imaging (fMRI) is currently the primary method for non-invasive functional localization in the brain. With the emergence of MR systems with field strengths of 4 Tesla and above, neuronal activation may be studied with unprecedented accuracy. In this article we present different approaches to use the improved sensitivity and specificity for expanding current fMRT resolution limits in space and time based on several 7 Tesla studies. In addition to the challenges that arise with ultra-high magnetic fields possible solutions will be discussed. PMID- 20012004 TI - [Corpus callosum. Landmark of the origin of cerebral diseases]. AB - Diseases of the corpus callosum include developmental disorders, immunomodulated CNS diseases, vascular malformations, disturbances of metabolism including the electrolyte homeostasis, secondary degenerations and mechanical injuries. This report provides information on the differential diagnosis of reversible and irreversible pathological changes of the corpus callosum with special focus on the localization, which often allows conclusions on the pathogenesis to be drawn. PMID- 20012006 TI - [Gastrointestinal stromal tumor (GIST) of the anterior rectal wall. R0 resection with simultaneous radical retropubic prostatectomy]. AB - This case report deals with a 79-year-old patient with a gastrointestinal stromal tumor (GIST) of the anterior rectal wall which was unusually located between the rectum and the prostate gland. In addition, this patient suffered from subvesical obstruction accompanied by an elevated PSA level. These circumstances led to our decision to operate on the tumor via simultaneous radical retropubic prostatectomy. In our opinion this resection technique was easier and less traumatic for the patient compared to procedures performed via the abdomen and perineum. This case report demonstrates that in the case of tumors located between the rectum and the prostate gland the differential diagnosis should include not only prostate carcinoma but also rare tumor entities such as GIST. PMID- 20012007 TI - [Ureteric reconstruction with reconfigured ileal segments according to Yang Monti. A 4-year prospective report]. AB - BACKGROUND: We report on 4 years experience with ileal ureteric replacement using the Yang-Monti procedure. PATIENTS AND METHODS: From April 2001 to January 2009 reconfigured ileal segments were used for total (in 16) or partial (in 2) substitution of the ureter in 18 patients (mean age 47.4 years) with functional ureteric loss secondary to radiogenic or iatrogenic conditions. An antireflux implantation into the native bladder was done in 16 patients. All patients were followed prospectively according to a standardized protocol. RESULTS: The mean follow-up was 4.2 years (0.5-8 years). There were no perioperative deaths. Ultrasound controls showed an improvement of the upper tract dilatation in 11, a constant finding in 5 and a worsening in 2 cases. All of the treated renal units had evidence of improved renal function in ten and stabilization in eight patients. Neither a metabolic complication nor mucous obstruction was observed. Minor short-term complications, mainly febrile urinary tract infection and paralytic ileus, occurred in 50% and long-term complications, infections and hernia in 22%. CONCLUSIONS: The ileal ureteral substitute with reconfigured segments offers distinct advantages. A short bowel segment is used with the consequent absence of metabolic complications and excessive mucous production. It allows construction of an ileal ureter with a suitable cross-sectional diameter without any need for tailoring and makes it possible to use an antireflux technique. The intermediate results are encouraging. PMID- 20012008 TI - [The patient with diabetes in urologic practice: a special risk for lower urinary tract symptoms? Results of the Witten diabetes survey of 4071 type 2 diabetics]. AB - Due to demographic developments in industrial nations, age-correlated diseases such as diabetes are becoming more important. In conclusion, lower urinary tract symptoms (LUTS) are a frequent reason for searching treatment by urologists. The association between LUTS and benign prostatic hyperplasia or neurologic diseases is accepted. In contrast there is limited knowledge about the connection of diabetes mellitus and LUTS although an autonomic neuropathy due to diabetes makes urinary bladder dysfunction likely. The following survey covered data of 4071 patients with diabetes type 2 with a medium age of 67,4 years and a diabetes duration of 8,8 years. The incidence of LUTS in diabetic patients increases by increasing age, longer diabetes duration and concomitant diabetes complications e. G. erectile dysfunction. There was a difference in severity of LUTS in patients in urologic treatment or in treatment of a general practitioner: 81,1% of the patients in urologic treatment reported LUTS; in comparison 60,7% of patients being treated by general practitioners complained of LUTS. The LUTS being reported in urologic practice were more severe; e. G. the number of incontinence episodes per week of urologic patients was 7,5, the number of incontinence episodes in general practice 5,7. Lower urinary tract symptoms were most frequently classified as OAB in male patients and classified as mixed incontinence in females. The amount of patients with LUTS was higher in patients with diabetes complications (79,8% vs. 49,9%); there was only a small difference in the number of patients with LUTS stratified by co-medication (64% patients with LUTS without any co-medication; 72% patients with co-medication). The high number of diabetic patients with LUTS in general and urologic practice indicates an underestimation of the problem. LUTS in patients with type-2-diabetes should be screened as eye complications, erectile dysfunction or circulatory disorders by urologists and general practitioners. PMID- 20012009 TI - Symptoms of depression but not anxiety are associated with central obesity and cardiovascular disease in people with type 2 diabetes: the Edinburgh Type 2 Diabetes Study. AB - AIMS/HYPOTHESIS: The aim of the study was to identify risk factors for depression and anxiety in a well-characterised cohort of individuals with type 2 diabetes mellitus. METHODS: We used baseline data from participants (n = 1,066, 48.7% women, aged 67.9 +/- 4.2 years) from the Edinburgh Type 2 Diabetes Study. Symptoms of anxiety and depression were assessed using the Hospital Anxiety and Depression Scale (HADS). Obesity was characterised according to both overall (body mass index, fat mass) and abdominal (waist circumference) measurements. Cardiovascular disease was assessed by questionnaire, physical examination and review of medical records. Stepwise multiple linear regression was performed to identify explanatory variables related to either anxiety or depression HADS scores. RESULTS: Abdominal obesity (waist circumference) and cardiovascular disease (ischaemic heart disease and ankle-brachial pressure index) were related to depression but not anxiety. Lifetime history of severe hypoglycaemia was associated with anxiety. Other cardiovascular risk factors or microvascular complications were not related to either anxiety or depressive symptoms. CONCLUSIONS/INTERPRETATION: Depression but not anxiety is associated with abdominal obesity and cardiovascular disease in people with type 2 diabetes mellitus. This knowledge may help to identify depressive symptoms among patients with type 2 diabetes who are at greatest risk. PMID- 20012010 TI - IL6 as a mediator of insulin resistance: fat or fiction? PMID- 20012011 TI - The metabolic syndrome: useful concept or clinical tool? Report of a WHO Expert Consultation. AB - This article presents the conclusions of a WHO Expert Consultation that evaluated the utility of the 'metabolic syndrome' concept in relation to four key areas: pathophysiology, epidemiology, clinical work and public health. The metabolic syndrome is a concept that focuses attention on complex multifactorial health problems. While it may be considered useful as an educational concept, it has limited practical utility as a diagnostic or management tool. Further efforts to redefine it are inappropriate in the light of current knowledge and understanding, and there is limited utility in epidemiological studies in which different definitions of the metabolic syndrome are compared. Metabolic syndrome is a pre-morbid condition rather than a clinical diagnosis, and should thus exclude individuals with established diabetes or known cardiovascular disease (CVD). Future research should focus on: (1) further elucidation of common metabolic pathways underlying the development of diabetes and CVD, including those clustering within the metabolic syndrome; (2) early-life determinants of metabolic risk; (3) developing and evaluating context-specific strategies for identifying and reducing CVD and diabetes risk, based on available resources; and (4) developing and evaluating population-based prevention strategies. PMID- 20012012 TI - Determinants of glucose tolerance in impaired glucose tolerance at baseline in the Actos Now for Prevention of Diabetes (ACT NOW) study. AB - AIMS/HYPOTHESIS: The aim of the study was to examine the determinants of oral glucose tolerance in 602 persons with impaired glucose tolerance (IGT) who participated in the Actos Now for Prevention of Diabetes (ACT NOW) study. METHODS: In addition to the 602 IGT participants, 115 persons with normal glucose tolerance (NGT) and 50 with impaired fasting glucose (IFG) were identified during screening and included in this analysis. Insulin secretion and insulin sensitivity indices were derived from plasma glucose and insulin during an OGTT. The acute insulin response (AIR) (0-10 min) and insulin sensitivity (S(I)) were measured with the frequently sampled intravenous glucose tolerance test (FSIVGTT) in a subset of participants. RESULTS: At baseline, fasting plasma glucose, 2 h postprandial glucose (OGTT) and HbA(1c) were 5.8 +/- 0.02 mmol/l, 10.5 +/- 0.05 mmol/l and 5.5 +/- 0.04%, respectively, in participants with IGT. Participants with IGT were characterised by defects in early (DeltaI (0-30)/DeltaG (0-30) x Matsuda index, where DeltaI is change in insulin in the first 30 min and DeltaG is change in glucose in the first 30 min) and total (DeltaI(0-120)/DeltaG(0-120) x Matsuda index) insulin secretion and in insulin sensitivity (Matsuda index and S(I)). Participants with IGT in whom 2 h plasma glucose was 7.8-8.3 mmol/l had a 63% decrease in the insulin secretion/insulin resistance (disposition) index vs participants with NGT and this defect worsened progressively as 2 h plasma glucose rose to 8.9-9.94 mmol/l (by 73%) and 10.0-11.05 mmol/l (by 80%). The Matsuda insulin sensitivity index was reduced by 40% in IGT compared with NGT (p < 0.005). In multivariate analysis, beta cell function was the primary determinant of glucose AUC during OGTT, explaining 62% of the variance. CONCLUSION: Our results strongly suggest that progressive beta cell failure is the main determinant of progression of NGT to IGT. PMID- 20012013 TI - A basic arthroscopy course based on motor skill training. AB - Aim of this study is to introduce, evaluate, and propose a basic arthroscopy training course with interactive methods as using bovine knees, joint models but mainly focusing on practices with motor skill-learning devices. This study presents the results of a skill training program which is based on "motor skill training" theory and using "bovine knees" to simulate a specific task as arthroscopy both in education and evaluation. The participants gave feedback regarding their expectations, fulfillment, and self-evaluation on the degree of their improvement levels, after which their skills were evaluated by an expert while they performed the arthroscopic procedure. During this evaluation of the task, a four-dimensional checklist which was prepared previously by the consensus of three experts was used, and a global grading of each rater was added at the end of the checklist. In this 2-day course, small group teaching sessions are integrated with active and experiential learning methods consisting of short presentations by the educators, demonstrations using joint models and audio visual teaching material, and significant continuous feedback by the teachers during the proceedings. Participants' expectations and post-course feedback results were obtained using a brief questionnaire which involved mostly open ended questions. The self-rating of "own basic diagnostic arthroscopy skill" on a scale of 0-10 was completed by the participants both previous to and following the course to establish a student-centered learning environment. Overall rating of the course was asked on a 1-5 (poor-excellent) Likert scale. The questioning of the participants who took part in the course during the last five terms involved a 100% feedback. The overall rating of the course was 4.36 +/- 0.47 on the 1-5 scale. The majority of the participants (90%) mentioned that the course met their expectations. The most beneficial activities were accepted to be the arthroscopic procedure applications on bovine knee, repetition of the manipulations under the supervision of the trainers, continuous feedback by experienced staff, and chance for immediate correction during the procedures (35/64). Pre- and post-course self-ratings of the participants inquiring about their ability to perform an arthroscopy alone showed significant improvement (4.2 vs. 7.7 P = 0.000, paired samples t test). All participants were found to be competent in the evaluation of their diagnostic arthroscopy skills on bovine knees. To design a skill teaching course based on the needs of the trainees, focusing on basic motor skill training exercises, and using bovine knee as a simulator is a safe, inexpensive, humanistic, and replicable method that proves a foundation for basic arthroscopic skills learning prior to patient encounter. PMID- 20012014 TI - Tunnel location in transparent 3-dimensional CT in anatomic double-bundle anterior cruciate ligament reconstruction with the trans-tibial tunnel technique. AB - It is difficult to precisely identify the locations of tunnels after double bundle anterior cruciate ligament (ACL) reconstruction postoperatively. Using our novel transparent 3-dimensional computed tomography (T-3DCT), we evaluated intra articular outlet locations and the angles of the anteromedial (AM) and posterolateral (PL) tunnels after anatomic double-bundle ACL reconstruction using the trans-tibial technique. A prospective study was performed with 123 consecutive patients. Tunnel outlet locations were identified on T-3DCT images showing the true lateral view of the femur and indicated by our originally defined X, Y coordinates. We also determined the angles between the tunnel axis and a joint surface line in the coronal plane, the long axis of the femur in the sagittal plane, and the posterior condyle line in the axial plane of both the femur and the tibia. The mean X, Y coordinates of the AM and PL tunnel outlets were 21, 43% and 0, 33%, respectively. In the coronal, sagittal, and axial planes, the mean AM femoral tunnel angles were 63 degrees, 48 degrees, and 55 degrees, respectively; the mean AM tibial tunnel angles in the tibia were 63 degrees, 49 degrees, and 71 degrees, respectively; the mean PL femoral tunnel angles were 38 degrees, 58 degrees, and 43 degrees, respectively; and the mean PL tibial tunnel angles were 46 degrees, 53 degrees, and 45 degrees, respectively. The AM and PL tunnel outlets and angles could be detailed precisely in three dimensions by using T-3DCT. This imaging technique may be useful to confirm surgical techniques and to improve clinical outcomes. PMID- 20012015 TI - Anatomy is the key. PMID- 20012016 TI - Treatment of articular cartilage lesions of the knee joint using a modified AMIC technique. AB - This study describes a modified AMIC technique consisting of perforations according to Pridie, rather than microfractures, and the covering of the focus of the lesion with a biological collagen patch enriched with bone marrow blood drawn through the knee itself. This technique allows advantages of both the Pridie technique and the in situ proliferation of mesenchymal cells beneath a biological collagen membrane, 'augmented', with bone marrow blood. The collagen membrane forms the roof of a 'biological chamber', and serves to protect and contains the stem cells as they differentiate into chondrocytes, which will form a healthy regenerative cartilage. PMID- 20012017 TI - The immediate effect of repeated loading on the compressive strength of young porcine lumbar spine. AB - The human spine is exposed to repeated loading during daily activities and more extremely during sports. Despite this, there remains a lack of knowledge regarding the immediate effects on the spine due to this mode of loading. Age specific spinal injury patterns has been demonstrated and this implies differences in reaction to load mode and load history The purpose of the present study was to investigate the impact of cyclic pre-loading on the biomechanical properties and fracture patterns of the adolescent spine in an experimental model. Eight functional spinal units from four young porcine spines were harvested. The functional spinal units were cyclic loaded with 20,000 cycles and then axially compressed to failure. The compression load at failure, ultimate stress and viscoelastic parameters were calculated. The functional spinal units were examined with plain radiography, computer tomography and MRI before and after the loading, and finally macroscopically and histologically. The median compression load at failure in this study was 8.3 kN (range 5.6-8.7 kN). The median deformation for all cases was 2.24 mm (range 2.30-2.7 mm) and stiffness was 3.45 N/mm (range 3.5-4.5 N/mm). A fracture was seen on radiograph in one case, on CT and macroscopically in seven, and on MRI and histologically in all eight cases. The cyclic loaded functional spinal units in the present study were not more sensitive to axial compression than non-cyclic loaded functional spinal units from young porcine. The endplate and the growth zone were the weakest part in the cyclic loaded functional spinal units. Disc signal reduction and disc height reduction was found on MRI. The E-modulus value found in this study was of the same order of magnitude as found by others using a porcine animal model. PMID- 20012018 TI - Fibroblast growth factor-23 is associated with C-reactive protein, serum phosphate and bone mineral density in chronic kidney disease. AB - We investigated the association between fibroblast growth factor-23 (FGF-23) and (1) the biochemical parameters implicated in chronic kidney disorder (CKD)-bone and mineral disorder (CKD-MBD) and (2) bone mineral density (BMD) in patients with CKD 1-4. C-reactive protein (CRP) and serum phosphate correlated with FGF 23. A significant association was seen between FGF-23 and BMD at the hip. INTRODUCTION: Circulating FGF-23 is elevated in CKD, although the primary stimulus remains unclear. Moreover, it is still unknown whether increase in FGF 23 has a biological effect on bone metabolism. The aim of the study was to investigate the association of FGF-23 with (1) the biochemical parameters linked with CKD-bone and mineral disorder (CKD-MBD) and (2) bone mineral density in CKD. METHODS: We studied 145 patients (74 M, 71 F) aged (mean [SD]) 53 [14] years with CKD 1-4. Serum calcium, phosphate, parathyroid hormone, FGF-23, 25 (OH) vitamin D, 1, 25 (OH)(2) vitamin D, bone turnover markers, CRP were determined. BMD was measured at the lumbar spine, femoral neck (FN), forearm, and total hip (TH). Multivariate analysis was undertaken to explore the association between (1) the biochemical variables and FGF-23 and (2) FGF-23 and BMD. RESULTS: Elevations in FGF-23 occurred in CKD stage 3 compared to CKD stage 1/2, although no significant differences in serum phosphate were observed. Serum phosphate (p<0.001), CRP (p<0.001) and diabetes mellitus (p<0.05) were associated with FGF-23. BMD Z-score was significantly lower at the TH and FN in CKD 4 (p<0.05). A significant association was seen between BMI, FGF-23, bone specific alkaline phosphatase and BMD at the TH (p<0.05). CONCLUSIONS: The data suggest that FGF-23 may be associated with parameters implicated in the complications of CKD. Longitudinal studies are required for further clinical evaluation. PMID- 20012019 TI - Educational achievement and fracture risk. PMID- 20012024 TI - Determination of brominated diphenyl ethers (from mono- to hexa- congeners) in indoor dust by pressurised liquid extraction with in-cell clean-up and gas chromatography-mass spectrometry. AB - This study presents a selective pressurised liquid extraction (PLE) and gas chromatography-mass spectrometry/mass spectrometry method for the determination of brominated diphenyl ethers (BDEs) in indoor dust. Selective PLE consisted of the addition of Florisil mixed with the sample in order to perform an in-cell clean-up. This approach provided a cleaner and almost colourless extract, ready to be injected in the gas chromatograph. The PLE conditions were studied using an experimental design, firstly a 4 x 3 x 2 multifactor categorical design to screen sorbent, solvent and temperature and then a central composite design to optimise sorbent mass, temperature and time. Finally, the number of extraction cycles was studied. The selected conditions were 4 g of Florisil all mixed with the sample and no additional clean-up sorbent layer, 1:1 n-hexane-dichloromethane, 60% flush volume, 40 degrees C, 1,500 psi, 2-min static time and one cycle. The proposed method allowed accurate determination of BDEs, with recovery values between 82% and 101% and detection limits between 0.06 and 0.24 ng g(-1). It also has advantages over other existing methods in terms of simplicity, automation, analysis time and solvent consumption. PMID- 20012021 TI - Neuropeptide Y (NPY) suppresses yohimbine-induced reinstatement of alcohol seeking. AB - INTRODUCTION: Reinstatement of responding to a previously alcohol-associated lever following extinction is an established model of relapse-like behavior and can be triggered by stress exposure. Here, we examined whether neuropeptide Y (NPY), an endogenous anti-stress mediator, blocks reinstatement of alcohol seeking induced by the pharmacological stressor yohimbine. MATERIALS AND METHODS: NPY [5.0 or 10.0 mug/rat, intracerebroventricularly (ICV)] dose-dependently blocked the reinstatement of alcohol seeking induced by yohimbine (1.25 mg/kg, i.p.) but failed to significantly suppress the maintenance of alcohol self administration. We then used c-fos expression mapping to examine neuronal activation following treatment with yohimbine or NPY alone or yohimbine following NPY pre-treatment. RESULTS AND DISCUSSION: The analysis was focused on a network of structures previously implicated in yohimbine-induced reinstatement, comprised of central (CeA) and basolateral (BLA) amygdala and the shell of the nucleus accumbens (Nc AccS). Within this network, both yohimbine and NPY potently induced neuronal activation, and their effects were additive, presumably indicating activation of excitatory and inhibitory neuronal populations, respectively. CONCLUSION: These results suggest that NPY selectively suppresses relapse to alcohol seeking induced by stressful events and support the NPY system as an attractive target for the treatment of alcohol addiction. PMID- 20012022 TI - Acute dopamine and/or serotonin depletion does not modulate mismatch negativity (MMN) in healthy human participants. AB - RATIONALE: Schizophrenia is commonly associated with impairments in pre-attentive change detection, as represented by reduced mismatch negativity (MMN). While the neurochemical basis of MMN has been linked to N-methyl-D: -aspartic acid (NMDA) receptor function, the roles of the dopaminergic and/or the serotonergic systems are not fully explored in humans. OBJECTIVES: The aim of the present study was to investigate the effects of acutely depleting dopamine (DA) and serotonin (5 hydroxytryptamine, 5-HT) alone or simultaneously by depleting their amino acid precursors on MMN in healthy participants. METHODS: Sixteen healthy male subjects participated in a double-blind, placebo-controlled, cross-over design in which each subject's duration MMN was assessed under four acute treatment conditions separated by a 5-day washout period: balanced amino acid control (no depletion), tyrosine/phenylalanine depletion (to reduce DA neurotransmission), tryptophan depletion (to reduce 5-HT neurotransmission) and tryptophan/tyrosine/phenylalanine depletion (to reduce DA and 5-HT neurotransmission simultaneously). RESULTS: Acute depletion of either DA and 5-HT alone or simultaneously had no effect on MMN. CONCLUSIONS: These findings suggest that modulation of the dopaminergic and serotonergic systems acutely does not lead to changes in MMN. PMID- 20012025 TI - An immunochemical test for rapid screening of zearalenone and T-2 toxin. AB - An immunochemically based test for non-instrumental simultaneous detection of zearalenone (ZEA) and T-2 toxin (T2) in feed was developed. The method combines clean-up of sample extract, pre-concentration of analytes by immunoextraction and immunodetection through the enzymatic reaction of horseradish peroxidase (HRP). The test is housed inside a standard 1-mL solid-phase extraction column and consists of three layers: two test layers (one for ZEA and another for T2) with immobilised specific antibodies and one control layer with bound anti-HRP antibodies. Feed extract was passed through an additional column with clean-up layer, which was disconnected after extract application. Total assay time was about 15 min for six samples and detection time was 4 min after chromogenic substrate application. Under optimised conditions a cut-off level for ZEA and T2 of 100 microg/kg was established. Different feed types were analysed for ZEA and T2 contamination by the proposed method and results were confirmed by LC-MS/MS. PMID- 20012026 TI - Stability of Se species in plant extracts rich in phenolic substances. AB - Since there is growing awareness of the strong dependence of the antioxidative function of selenium (Se) upon its chemical form, the stability of Se species during sample preparation is an important factor in obtaining qualitative and quantitative results. Many plant samples are rich in phenolic compounds (antioxidants), but data about their effect on specific Se species in extracts of plant samples are scarce. Therefore, the aim of this study was to investigate the effect of the most common phenolic substances in plant parts, namely tannin and the flavonoid rutin, on the concentration and/or transformation of several Se species (SeMet, SeCys(2), SeMeSeCys, Se(VI) and Se(IV)) during sample preparation (24 h incubation at 37 degrees C) and storage (4 days at 4 degrees C). Moreover, the effect observed was then studied in a real sample, buckwheat, because this plant is known as a rich source of phenolics, especially tannin and rutin. Se speciation was carried out by on-line coupling of ion-exchange HPLC-ICP-MS after water and enzymatic (protease) hydrolysis. The results showed that the ratio between the two antioxidants has an important role. When the antioxidants were present together, the response for Se(IV) was observed to start to decrease only at a ratio of rutin to tannin of 1:100 (w/w), indicating the ratio between antioxidants in buckwheat seeds. After water extraction, only 40% and after enzymatic extraction 80% of Se(IV) remained, but no other Se compound was detected with the system used. Furthermore, the extracts were not stable during storage at 4 degrees C. Signals for other Se species were stable. The results obtained for buckwheat seeds showed a decrease in Se(IV) response during sample preparation and storage, comparable to the one obtained with the experiments performed in vitro. However, Se species in extracts of other buckwheat parts (leaves, stems and sprouts) were stable. These results indicate that reactions in the extraction process and during storage may affect Se speciation and may result in misidentifications and inaccurate values. PMID- 20012027 TI - Increased efficacy for in-house validation of real-time PCR GMO detection methods. AB - To improve the efficacy of the in-house validation of GMO detection methods (DNA isolation and real-time PCR, polymerase chain reaction), a study was performed to gain insight in the contribution of the different steps of the GMO detection method to the repeatability and in-house reproducibility. In the present study, 19 methods for (GM) soy, maize canola and potato were validated in-house of which 14 on the basis of an 8-day validation scheme using eight different samples and five on the basis of a more concise validation protocol. In this way, data was obtained with respect to the detection limit, accuracy and precision. Also, decision limits were calculated for declaring non-conformance (>0.9%) with 95% reliability. In order to estimate the contribution of the different steps in the GMO analysis to the total variation variance components were estimated using REML (residual maximum likelihood method). From these components, relative standard deviations for repeatability and reproducibility (RSD(r) and RSD(R)) were calculated. The results showed that not only the PCR reaction but also the factors 'DNA isolation' and 'PCR day' are important factors for the total variance and should therefore be included in the in-house validation. It is proposed to use a statistical model to estimate these factors from a large dataset of initial validations so that for similar GMO methods in the future, only the PCR step needs to be validated. The resulting data are discussed in the light of agreed European criteria for qualified GMO detection methods. PMID- 20012028 TI - Evaluation of the state-of-the-art measurement capabilities for selected PBDEs and decaBB in plastic by the international intercomparison CCQM-P114. AB - An international intercomparison involving eight national metrology institutes (NMIs) was conducted to establish their current measurement capabilities for determining five selected congeners from the brominated flame retardant classes polybrominated diphenyl ethers and polybrominated biphenyls. A candidate reference material consisting of polypropylene fortified with technical mixtures of penta-, octa- and decabromo diphenyl ether and decabromo biphenyl, which was thoroughly assessed for material homogeneity and stability, was used as study material. The analytical procedures applied by the participants differed with regard to sample pre-treatment, extraction, clean-up, employed calibrants and type of calibration procedure as well as regarding analytical methods used for separation, identification and quantification of the flame retardant congeners (gas chromatography coupled to an electron capture detector (GC-ECD), gas chromatography-mass spectrometry in the electron ionisation mode (GC-EI-MS), gas chromatography-mass spectrometry in the electron capture negative ionisation mode (GC-ECNI-MS), and liquid chromatography-inductive coupled plasma-mass spectrometry (LC-ICP-MS)). The laboratory means agreed well with relative standard deviations of the mean of means of 1.9%, 4.8%, 5.5% and 5.4% for brominated diphenyl ether (BDE) 47, 183 and 209 and for the brominated biphenyl (BB) congener 209, respectively. For BDE 206, a relative standard deviation of 28.5% was obtained. For all five congeners, within-laboratory relative standard deviations of six measurements obtained under intermediate precision conditions were between 1% and 10%, and reported expanded measurements uncertainties typically ranged from 4% to 10% (8% to 14% for BDE 206). Furthermore, the results are in good agreement with those obtained in the characterization exercise for determining certified values for the flame retardant congeners in the same material. The results demonstrate the state-of-the-art measurement capabilities of NMIs for quantifying representative BDE congeners and BB 209 in a polymer. The outcome of this intercomparison (pilot study) in conjunction with possible improvements for employing exclusively calibrants with thoroughly assessed purity suggests that a key comparison aiming at underpinning calibration and measurement capability (CMC) claims of NMIs can be conducted. PMID- 20012029 TI - Stimulation of endothelial progenitor cells: a new putative effect of several cardiovascular drugs. AB - The role of vascular endothelium in cardiovascular disorders is well recognized. Mature endothelial cells contribute to the repair of endothelial injury, but they only have a limited capacity to do so. This has led to growing interest and further investigation into circulating endothelial progenitor cells (EPCs) and their role in vascular healing, repair, and postnatal neovascularization. The current perception of vascular health is that of a balance between ongoing injury and resultant vascular repair, mediated at least in part by circulating EPCs. Circulating EPCs play an important role in accelerating endothelialization at areas of vascular damage, and EPC enumeration is a viable strategy for assessing reparative capacity. Recent studies have shown that EPCs are affected both in number and function by several cardiovascular risk factors as well as various cardiovascular disease states, such as hypertension, hypercholesterolemia, and coronary artery disease. The present review summarizes the most relevant studies on the effects of cardiovascular drugs on vascular function and EPCs, focusing on their mechanisms of action. PMID- 20012032 TI - Certain non-standard coding tables appear to be more robust to error than the standard genetic code. AB - Since the identification of the Standard Coding Table as a "universal" method to translate genetic information into amino acids, exceptions to this rule have been reported, and to date there are nearly 20 alternative genetic coding tables deployed by either nuclear genomes or organelles of organisms. Why are these codes still in use and why are new codon reassignments occurring? This present study aims to provide a new method to address these questions and to analyze whether these alternative codes present any advantages or disadvantages to the organisms or organelles in terms of robustness to error. We show that two of the alternative coding tables, The Ciliate, Dasycladacean and Hexamita Nuclear Code (CDH) and The Flatworm Mitochondrial Code (FMC), exhibit an advantage, while others such as The Yeast Mitochondrial Code (YMC) are at a significant disadvantage. We propose that the Standard Code is likely to have emerged as a "local minimum" and that the "coding landscape" is still being searched for a "global" minimum. PMID- 20012030 TI - Hepatic CYP2A6 levels and nicotine metabolism: impact of genetic, physiological, environmental, and epigenetic factors. AB - PURPOSE: We investigated the role of genetic, physiological, environmental, and epigenetic factors in regulating CYP2A6 expression and nicotine metabolism. METHODS: Human livers (n = 67) were genotyped for CYP2A6 alleles and assessed for nicotine metabolism and CYP2A6 expression (mRNA and protein). In addition, a subset of livers (n = 18), human cryopreserved hepatocytes (n = 2), and HepG2 cells were used for DNA methylation analyses. RESULTS: Liver samples with variant CYP2A6 alleles had significantly lower CYP2A6 protein expression, nicotine C oxidation activity, and affinity for nicotine. Female livers had significantly higher CYP2A6 protein and mRNA expression compared to male livers. Livers exposed to dexamethasone and phenobarbital had higher CYP2A6 expression and activity, however the difference was not statistically significant. Age and DNA methylation status of the CpG island and a regulatory site were not associated with altered CYP2A6. CONCLUSIONS: We identified genotype, gender, and exposure to inducers as sources of variation in CYP2A6 expression and activity, but much variation remains to be accounted for. PMID- 20012033 TI - Diagnosis of ductal aneurysm using three-dimensional CT reconstruction. PMID- 20012034 TI - Fractures in infants and toddlers with rickets. AB - BACKGROUND: Rickets affects young infants and toddlers. However, there is a paucity of literature regarding the types of fractures that occur in rachitic patients. OBJECTIVE: To evaluate the age of patients at which radiographically evident rickets occurs, and to characterize the age incidence and fractures that are observed in infants and toddlers with radiographically evident rickets. MATERIALS AND METHODS: A retrospective study of children younger than 24 months was performed. Clinical data and radiographs were reviewed. Radiographs obtained within 1 month of the diagnosis were evaluated for the presence or absence of osteopenia, presence or absence of fraying-cupping, and presence and characterization of fractures. RESULTS: After exclusion criteria were applied, 45 children were included in the study. Children with rickets evident by radiograph were in the age range of 2-24 months. Fractures were present in 17.5% of the study group, exclusively in mobile infants and toddlers. Fracture types included transverse long bone fractures, anterior and anterior-lateral rib fractures, and metaphyseal fractures. All fractures occurred exclusively in patients with severe, overtly evident rickets. CONCLUSION: Fractures occur in older infants and toddlers with overt rickets and can be seen by radiograph. Fractures do not resemble high-risk non-accidental trauma fractures. PMID- 20012040 TI - Free vascularised fibular grafting with OsteoSet(r)2 demineralised bone matrix versus autograft for large osteonecrotic lesions of the femoral head. AB - The aim of this study was to compare the safety and efficacy of OsteoSet(r)2 DBM with autologous cancellous bone in free vascularised fibular grafting for the treatment of large osteonecrotic lesions of the femoral head. Twenty-four patients (30 hips) with large osteonecrotic lesions of the femoral head (stage IIC in six hips, stage IIIC in 14, and stage IVC in ten, according to the classification system of Steinberg et al.) underwent free vascularised fibular grafting with OsteoSet(r)2 DBM. This group was retrospectively matched to a group of 24 patients (30 hips) who underwent free vascularised fibular grafting with autologous cancellous bone during the same time period according to the aetiology, stage, and size of the lesion and the mean preoperative Harris hip score. A prospective case-controlled study was then performed with a mean follow up duration of 26 months. The results show no statistically significant differences between the two groups in overall clinical outcome or the radiographic assessment. Furthermore, no adverse events related to the use of the OsteoSet(r)2 DBM were observed. The results demonstrate that OsteoSet(r)2 DBM combined with autograft bone performs equally as well as that of autologous bone alone. Therefore, OsteoSet(r)2 DBM can be used as a safe and effective graft extender in free vascularised fibular grafting for large osteonecrotic lesions of the femoral head. PMID- 20012039 TI - Ulnar-sided wrist pain. II. Clinical imaging and treatment. AB - Pain at the ulnar aspect of the wrist is a diagnostic challenge for hand surgeons and radiologists due to the small and complex anatomical structures involved. In this article, imaging modalities including radiography, arthrography, ultrasound (US), computed tomography (CT), CT arthrography, magnetic resonance (MR) imaging, and MR arthrography are compared with regard to differential diagnosis. Clinical imaging findings are reviewed for a more comprehensive understanding of this disorder. Treatments for the common diseases that cause the ulnar-sided wrist pain including extensor carpi ulnaris (ECU) tendonitis, flexor carpi ulnaris (FCU) tendonitis, pisotriquetral arthritis, triangular fibrocartilage complex (TFCC) lesions, ulnar impaction, lunotriquetral (LT) instability, and distal radioulnar joint (DRUJ) instability are reviewed. PMID- 20012041 TI - Pure subdural hematoma due to cerebral aneurysmal rupture: an often delayed diagnosis. PMID- 20012042 TI - Visibility of the lateral collateral ligaments in routine computed tomography of the ankle. PMID- 20012043 TI - Short and long-term outcome of treatment with high-dose melphalan and stem cell transplantation for multiple myeloma-associated AL amyloidosis. AB - High-dose melphalan chemotherapy and autologous peripheral blood stem cell transplantation (HDM/SCT) has been shown to result in a durable hematologic response and prolonged overall survival in systemic amyloid light-chain (AL) amyloidosis as well as multiple myeloma. However, little is known about the myeloma associated with AL amyloidosis (MM/AL). In this retrospective study, we evaluated 87 patients with MM/AL from 1994 to 2007. Sixteen of these patients underwent HDM/SCT at Boston University Medical Center. Three patients (19%) died from treatment-related mortality. The overall median survival for all 87 patients was 22 months by Kaplan-Meier estimates. However, this was improved to 54.5 months for those who received HDM/SCT compared to 21 months for those who did not receive HDM/SCT. A hematologic complete response was achieved by 25% (4/16) of patients at 6 months after HDM/SCT. Hematologic relapses occurred in 60% of patients at a median of 1 year after HDM/SCT. In conclusion, HDM/SCT can prolong overall survival in patients with MM/AL who are eligible to receive it. PMID- 20012044 TI - Hematoma of the iliopsoas muscle due to thrombocytopenia resulting from the administration of a third-generation cephalosporin. PMID- 20012045 TI - Bortezomib, thalidomide, dexamethasone induction therapy followed by melphalan, prednisolone, thalidomide consolidation therapy as a first line of treatment for patients with multiple myeloma who are non-transplant candidates: results of the Korean Multiple Myeloma Working Party (KMMWP). AB - Bortezomib (VELCADE), thalidomide and dexamethasone (VTD), as well as melphalan, prednisolone, and thalidomide (MPT) therapy, are highly effective in patients with multiple myeloma. We evaluated the responses and survival times of 35 patients treated with VTD followed by MPT. All patients were newly diagnosed and non-transplantation candidates. Patients received six cycles of VTD, which were followed by eight cycles of MPT. Approximately 97% of patients exhibited early responses to therapy, as early as the second cycle of VTD. Thirty percent of the responses were high quality, which was defined as a complete response (CR), a near-CR or a very good partial response. High-risk patients were defined as patients with any of the following aberrations: del(13), t(4;14), or del(17p). The remaining patients were defined as standard risk. Eleven high-risk patients showed 100% response rates, including 91% high-quality responses. In contrast, 13 standard-risk patients exhibited 92% response rates, including 61% high-quality responses. The overall 2-year survival rates were 60% in high-risk patients and 85% in standard-risk patients, which was not significantly different. As a first line therapy, VTD followed by MPT has the potential to provide high-quality responses with durable remission among elderly and high-risk patients (clinicaltrials.gov identifier: NCT00320476). PMID- 20012046 TI - Attraction of pinewood nematode to endoparasitic nematophagous fungus Esteya vermicola. AB - The investigations on attraction of nematodes to nematophagous fungi have mostly dealt with the nematode-trapping species. Esteya vermicola is the endoparasitic fungus of pinewood nematode (PWN) with high infection activity. In the present study, the attraction of PWNs to E. vermicola was investigated. It was confirmed that the living mycelia and exudative substances of E. vermicola were attractive to PWN. Compared with the nematode-trapping fungus A. brochopaga as well as nematode-feeding fungus B. cinerea, E. vermicola showed the significantly strongest attraction ability to nematode. It therefore appeared that the attraction ability reflects the dependence of the fungi on nematodes for nutrients. Furthermore, a new method was developed and used in the study to confirm the effect of volatile substances for the attraction of nematode to fungi. The results suggested that the attractive substances were consisted of avolatile exudative and volatile diffusing compounds. PMID- 20012047 TI - Stochastic modelling of biased cell migration and collagen matrix modification. AB - Matrix dynamics plays a crucial role in several physiological and pathological processes. In this paper we develop a model framework, which describes the temporal fibre network evolution depending on the influence of migrating fibroblasts. The cells are regarded as discrete objects in the plane, whose velocities are determined by a generalised Langevin equation. For its solution we verify existence and uniqueness. The courses of the trajectories are affected by two external impulses, chemotaxis and contact guidance, respectively. The extracellular matrix is described by a continuous vector field which contains both information on density and orientation of the fibrous material. Modelling dynamic interaction between the discrete and the continuum variables is an essential point of this paper. In particular, the smoothing of the fluctuating paths plays a key role. Besides a detailed description of the formulated equations, we also supply the condensed pseudo code of the algorithm. We investigate several examples and present results both from artificial and real data. PMID- 20012048 TI - Dietary flavonoid apigenin inhibits endothelin-1-induced contraction of collagen gel. PMID- 20012049 TI - Late response to anti-TNF-alpha therapy in refractory mucocutaneous lesions of Behcet's disease. AB - Behcet's disease (BD) is a multisystem chronic inflammatory disorder characterized by oral and genital ulceration and ocular involvement. Recurrent oral and genital ulcers are the most common symptoms of BD and occur in more than 80% of patients. The treatments of these disease manifestations include colchicine, corticosteroids and immunosuppressive drugs in severe cases. Anti-TNF alpha therapy may be useful in refractory severe BD, particularly for ocular, central nervous system, gastrointestinal and refractory mucocutaneous lesions. During a 2-year period, 280 patients suffering from rheumatic diseases received anti-TNF-alpha agents at the infusion center of our University Hospital. Two patients (0.7%) presented BD; one of them had celiac disease as well, with recalcitrant mucocutaneous lesions that were not responsive to immunosuppressive drugs. We reported those patients who were successfully treated with infliximab and adalimumab, despite their late response. PMID- 20012050 TI - A case of granulomatous mastitis with erythema nodosum and arthritis. AB - Idiopathic granulomatous mastitis is an inflammatory mass in the breast. Patients typically present with a firm breast mass, often associated with inflammation of the overlying skin. Nipple retraction, peau d'orange-like changes, and axillary adenopathy may be present. We describe a 23-year-old pregnant woman with arthritis, erythema nodosum, and mastitis. PMID- 20012051 TI - The effects of electrical stimulation combined with continuous passive motion versus isometric exercise on symptoms, functional capacity, quality of life and balance in knee osteoarthritis: randomized clinical trial. AB - The aim of this study was to evaluate effects of electrical stimulation combined with continuous passive motion (CPM-ES) versus isometric exercise on symptoms, functional capacity, quality of life, muscle strength, knee and thigh circle measurements, and balance in knee osteoarthritis (OA). This is a randomized clinical trial. The study was done in Gulhane Military Medical Academy (GMMA) Rehabilitation Center. Forty patients with knee OA were included in this study. Participants were randomly assigned to two groups: 20 patients placed in Group 1 were treated with conventional physical therapy and CPM-ES combination; 20 patients in Group 2 were treated with conventional physical therapy and isometric exercise. Therapies were applied 3 weeks, 5 days per week. The following main outcome measures were done: values of pain (VAS was used), WOMAC, SF-36, knee and thigh circle measurements, isokinetic tests, dynamic and static balance tests were determined at baseline and after the treatment. There were no statistically significant differences in the tested variables between the groups for post treatment values. Dynamic and static balance test improved statistically strongly significantly in both groups. The findings of this study demonstrate that knee OA patients could improve their balance function in both static and dynamic conditions after CPM-ES combination or isometric exercise therapy. The improvement might prevent knee OA patients from falling down and increase their sense of security during physical activities. PMID- 20012052 TI - Pulmonary hypertension in systemic lupus erythematosus: relationship with antiphospholipid antibodies and severe disease outcome. AB - Pulmonary hypertension (PH) in systemic lupus erythematosus (SLE) is associated with an unfavorable prognosis. We investigated the characteristics of SLE patients with PH. The patients with a pulmonary artery systolic pressure more than 30 mmHg at rest on echocardiogram were diagnosed with PH. Echocardiography was done only in patients with clinical or radiological evidence suggesting PH. Right heart catheterization was not performed. We identified 10 SLE patients with PH between 1980 and 2000. We compared their clinical and laboratory parameters with those of 97 consecutive SLE patients without PH. Nine of the ten patients with PH were females. The mean age at the time of SLE onset was 25.2 +/- 6.9 years; the mean duration of follow-up was 93.4 +/- 52.8 months, and the interval between the onset of SLE and PH diagnosis was 9.0 +/- 4.6 (5-21) years. Antiphospholipid antibody positivity was significantly higher in the PH group (80 vs. 36%; p < 0.05), but there was no significant difference between two groups in regard to secondary antiphospholipid syndrome. The frequency of Raynaud's phenomenon was higher in PH group (60 vs. 27%; p < 0.05). Renal involvement (80 vs. 43%; p < 0.05), neuropsychiatric involvement (40 vs. 7.2%; p < 0.005) and serositis (70 vs. 14.4%; p < 0.001) were significantly more frequent in the PH group. The mean damage score in patients with and without PH were 4.0 +/- 2.4 and 0.4 +/- 1.0, respectively (p < 0.001). Four patients with PH died during the follow-up. This study reveals that the presence of PH defines a subgroup of patients with a severe disease and increased mortality. Antiphospholipid antibodies and Raynaud's phenomenon may contribute to the pathogenesis of PH. We recommend that all patients with SLE, especially those positive for antiphospholipid antibodies and/or with signs of Raynaud's phenomenon should be regularly evaluated for the development of PH. PMID- 20012053 TI - T-cell responses to versican in ankylosing spondylitis. AB - The objective of our study was to undertake a systematic analysis of the T-cell response to the proteoglycan versican G1-globular domain (VG1) in ankylosing spondylitis (AS) as immunity to VG1 in mice can induce a pathology closely resembling AS. Peripheral blood lymphocytes from 36 AS patients and 33 healthy controls were incubated with recombinant human VG1 in culture for 6 h. T-cell responses were assessed by FACS analyses using mAb against surface expression of the activation marker CD69 and against the intracellular cytokines interferon (IFN)-gamma, tumor necrosis factor (TNF)-alpha. T cells activated by exposure to versican were determined by assessing the percentage of CD4+ or CD8+ T cells that were CD69/cytokine double-positive cells as compared to isotype control staining. In the AS patients, exposure to VG1 resulted in increased expression by CD4+ T cells of IFN-gamma in 55.6% of patients and of TNF-alpha in 52.8% of patients. In the controls, only 36.4% of subjects demonstrated an IFN-gamma response and 36.4% demonstrated a TNF-alpha response (P value 0.148, 0.227, respectively). With respect to CD8+ T-cell responses, versican stimulation enhanced IFN-gamma expression in 44.4% of AS patients and 39.4% of controls, and enhanced TNF-alpha response in 50.0% of AS patients and 39.4% of controls (P value 0.620, 0.327, respectively). Although, there was no statistically significant difference in the magnitude of the IFN-gamma or TNF secretion by CD4+ T cells and CD8+ T cells between AS and controls, our results demonstrate an enhanced T-cell response to VG1 in AS. PMID- 20012054 TI - A case of juvenile idiopathic polyarticular arthritis complicated by IgA deficiency in 22q11 deletion syndrome. AB - Chronic arthritis may occur in association with antibody deficiency and chromosomal aberrations. This report presents the case of a 6-year-old girl with chromosome 22q11 deletion syndrome and chronic arthritis. The onset of arthritis occurred at 4 years of age. The chronic arthritis course has been the polyarticular type. Neither antinuclear antibody nor rheumatoid factor was detected. Serum IgA was extremely low. She was diagnosed with juvenile idiopathic polyarticular arthritis (JIA) complicated by IgA deficiency in the 22q11 deletion syndrome. There is an increased prevalence of chronic arthritis in association with 22q11 deletion syndrome with IgA deficiency, but the reasons for this association are unknown. This study evaluated the possible correlation between cytokines and the susceptibility to chronic arthritis in the 22q11 deletion syndrome with IgA deficiency. The expression of pro-inflammatory cytokines such as IL-8, IL-6, MIP-1beta, and MCP-1 suggests that T and B cells, macrophages and neutrophils modulate joint inflammation by an immune response. And the presence of IL-10 and IL-5 might suggest that the synovitis is associated with JIA and IgA deficiency. PMID- 20012055 TI - Differences in osmotolerance in freshwater and brackish water populations of Theodoxus fluviatilis (Gastropoda: Neritidae) are associated with differential protein expression. AB - The euryhaline gastropod Theodoxus fluviatilis is found in northern Germany in freshwater or in brackish water habitats in the Baltic Sea. Previous studies have revealed that individuals from both habitats are not distinguishable by morphological characters or by sequence comparison of DNA encoding 16S RNA or cytochrome C. As reported in this study, animals collected in the two habitats differ substantially in their physiological ability to adapt to different salinities. Comparison of accumulation rates of ninhydrin-positive substances (NPS) in foot muscle upon transfer of animals to higher medium salinities revealed that brackish water animals were perfectly able to mobilize NPS, while freshwater animals had only limited ability to do so. In an attempt to explore whether this difference in physiology may be caused by genetic differentiation, we compared protein expression patterns of soluble foot muscle proteins using 2D gel electrophoresis and silver staining. Of the 40 consistently detected protein spots, 27 showed similar levels in protein expression in animals collected from freshwater or brackish water habitats, respectively. In 12 spots, however, protein concentration was higher in brackish water than in freshwater animals. In four of these spots, expression levels followed increases or decreases in medium salinities. In a different set of 4 of these 12 spots, protein levels were always higher in brackish water as compared to freshwater animals, regardless of their physiological situation (14 days in artificial pond water or in medium with a salinity of 16 per thousand). The remaining 4 of the 12 spots had complex expression patterns. Protein levels of the remaining single spot were generally higher in freshwater animals than in brackish water animals. These expression patterns may indicate that freshwater and brackish water animals of T. fluviatilis belong to different locally adapted populations with subtle genetic differentiation. PMID- 20012056 TI - The vitellogenin of the bumblebee, Bombus hypocrita: studies on structural analysis of the cDNA and expression of the mRNA. AB - In this present study, the cDNA of Bombus hypocrita vitellogenin (Vg) was cloned and sequenced. It is composed of 5,478 bp and contains an ORF of 1,772 amino acids within a putative signal peptide of 16 residues. The deduced amino acid sequence shows significant similarity with Bombus ignitus (95%) and Apis mellifera (52%) and a high number of conserved motifs. Close to the C terminus there is a GL/ICG motif followed by nine cysteines, and a DGXR motif is located 18 residues upstream from the GL/ICG motif. Moreover, we predicted the 3D structure of B. hypocrita Vg. Furthermore, the Vg mRNA of B. hypocrita was spatio temporally analyzed in different castes (such as queen, worker and drone) from pupae to adult. The Vg mRNA was found in the white-eyed pupal (Pw) stage in queens, and the expression increased during the entire pupal development and attained its peak in the dark brown pupal stage. It also had a high expression in the adult fat body. In workers, the Vg expression was detected in the Pw stage, and its levels increased with age with the highest in 15 days. Afterward, it decreased progressively. Vg mRNA was also observed in drones, with a higher level of expression shown in only freshly molted adult drones. PMID- 20012057 TI - Reflex bradycardia does not influence oxygen consumption during hypoxia in the European eel (Anguilla anguilla). AB - Most teleost fish reduce heart rate when exposed to acute hypoxia. This hypoxic bradycardia has been characterised for many fish species, but it remains uncertain whether this reflex contributes to the maintenance of oxygen uptake in hypoxia. Here we describe the effects of inhibiting the bradycardia on oxygen consumption (MO(2)), standard metabolic rate (SMR) and the critical oxygen partial pressure for regulation of SMR in hypoxia (Pcrit) in European eels Anguilla anguilla (mean +/- SEM mass 528 +/- 36 g; n = 14). Eels were instrumented with a Transonic flow probe around the ventral aorta to measure cardiac output (Q) and heart rate (f (H)). MO(2) was then measured by intermittent closed respirometry during sequential exposure to various levels of increasing hypoxia, to determine Pcrit. Each fish was studied before and after abolition of reflex bradycardia by intraperitoneal injection of the muscarinic antagonist atropine (5 mg kg(-1)). In the untreated eels, f (H) fell from 39.0 +/ 4.3 min(-1) in normoxia to 14.8 +/- 5.2 min(-1) at the deepest level of hypoxia (2 kPa), and this was associated with a decline in Q, from 7.5 +/- 0.8 mL min(-1) kg(-1) to 3.3 +/- 0.7 mL min(-1) kg(-1) in normoxia versus deepest hypoxia, respectively. Atropine had no effect on SMR, which was 16.0 +/- 1.8 mumol O(2) kg(-1) min(-1) in control versus 16.8 +/- 0.8 mumol O(2) kg(-1) min(-1) following treatment with atropine. Atropine also had no significant effect on normoxic f (H) or Q in the eel, but completely abolished the bradycardia and associated decline in Q during progressive hypoxia. This pharmacological inhibition of the cardiac responses to hypoxia was, however, without affect on Pcrit, which was 11.7 +/- 1.3 versus 12.5 +/- 1.5 kPa in control versus atropinised eels, respectively. These results indicate, therefore, that reflex bradycardia does not contribute to maintenance of MO(2) and regulation of SMR by the European eel in hypoxia. PMID- 20012058 TI - Dimensions of the craniocervical junction in longitudinal analysis of normal growth. AB - INTRODUCTION: Abnormal configuration of the craniocervical junction in the form of basilar impression or invagination, with often-associated platybasia, is a clinically significant cause of neurological symptoms particularly in patients with inherited diseases affecting the connective tissue. To better understand the course of development of these basilar abnormalities and further define their diagnostic criteria in children, we analysed longitudinally changes in the vertical dimensions of the craniocervical junction and in the flexion of the anterior skull base in normal growing individuals. METHODS: The distance of the odontoid process to four reference lines and the anterior skull base angle was measured in consecutive series of at least five lateral skull radiographs of 30 females and 23 males. Their mean age was 6.4 years at the beginning and 22.4 years at the end of the observation period. RESULTS: In young children, the odontoid process was situated in a caudal relation to the skull base structures and reached a level similar to that of adults approximately at the age of 7 years in both males and females. Cross-sectional observation of the results camouflages, however, how the intra-individual changes were markedly non-linear. Changes in the anterior skull base angle remained nonsignificant. CONCLUSIONS: Normal values for McRae's, Chamberlain's and McGregor's measurements and the more recently documented D-M measurement are age dependent. We provide reference values specific for ages from 4 years. A notable deviation from the documented values indicates a need of further examination. PMID- 20012059 TI - Beta1 integrin and VEGF expression in an experimental model of brain tissue heterotopia in the lung. AB - INTRODUCTION: Integrins and vascular endothelial growth factor (VEGF) are crucially involved in interaction, proliferation, migration, and survival of the cells. However, there is no report in the literature about beta1 integrin and VEGF expression in heterotopic brain tissue. PURPOSE: The aim of this study was to assess beta1 integrin and VEGF expression in experimental brain tissue heterotopia in the lung during both fetal and neonatal periods. MATERIALS AND METHODS: Twenty-four pregnant female Swiss mice were used to induce brain tissue heterotopia on the 15th gestational day. Briefly, the brain of one fetus of each dam was extracted, disaggregated, and injected into the right hemithorax of siblings. Six of these fetuses with pulmonary brain tissue implantation were collected on the 18th gestational day (group E18) and six other on the eighth postnatal day (group P8). RESULTS: Immunohistochemistry of the fetal trunks showed implantation of glial fibrillary acidic protein- and neuronal nuclei positive heterotopic brain tissue, which were also positive for beta1 integrin and VEGF in both groups E18 and P8. CONCLUSION: These results indicate that brain tissue heterotopia during fetal and postnatal period is able to complete integration with the lung tissue as well as to induce vascular proliferation which are the necessary steps for a successful implantation. PMID- 20012061 TI - Sylvian fissure arachnoid cysts: we do operate on them but should it be done? PMID- 20012060 TI - Endoscopic anatomy of the oculomotor nerve: defining the blind spot during endoscopic skull base surgery. AB - OBJECTIVE: The objective of this study is to define the endoscopic anatomy of the oculomotor nerve (CN III) and its neurovascular relations in order to facilitate surgical procedures and avoid injury to this nerve during endoscopic endonasal approach to the skull base. MATERIALS AND METHODS: Endoscopic anatomy of the cavernous sinus was studied in seven fresh adult cadavers bilaterally and the basal cisterns in five fresh adult cadavers. Extended endoscopic endonasal suprasellar approach was performed to expose the oculomotor nerve in the interpeduncular cistern and the endoscopic endonasal transethmoidopterygoidosphenoidal approach to expose the oculomotor nerve within the cavernous sinus. RESULTS: The extraorbital part of the oculomotor nerve can be divided into three segments in regard to the cisterns and venous spaces that are being transected: the interpeduncular segment, the cisternal segment, and the intercavernous segment. Of these segments, only the cisternal segment could not be exposed since this segment was located at the initial part of the roof of the cavernous sinus, anterolateral to the posterior clinoid, and posteroinferior to the anterior clinoid processes. Thus, cisternal segment of the oculomotor nerve was considered a blind spot during endoscopic approaches to the skull base. CONCLUSION: We defined the endoscopic anatomy of the CN III and the related neurovascular structures and proposed a new segmental classification of extraorbital oculomotor nerve. Awareness of the endoscopic anatomy and the new segmental classification of the CN III may prove helpful in avoiding the risk of nerve injury during endoscopic endonasal surgery for skull base pathologies. PMID- 20012062 TI - Upregulation of mir-221 and mir-222 in atypical teratoid/rhabdoid tumors: potential therapeutic targets. AB - PURPOSE: The aim of this study is to search for new therapeutic targets for atypical teratoid-rhabdoid tumors (ATRT). METHODS: To achieve this, we compared the expression of 365 microRNAs among ATRT, medulloblastomas, and normal brain. RESULTS: MiR-221 and miR-222 were within the top differentially expressed microRNAs. The deregulated expression of miR221/222 was demonstrated to inhibit the expression of the tumor suppressor and inhibitor of cell cycle p27(Kip1). Here, we demonstrated the negative regulation of p27(Kip1) by miR-221/222 in ATRT using microarray, real-time reverse transcriptase polymerase chain reaction, and immunohistochemistry. CONCLUSION: As anti-miR therapy was recently proposed as an alternative treatment for cancer, these findings suggest that anti-miR-221/222 therapy might have therapeutic potential in ATRT. PMID- 20012063 TI - Humor as a character strength among the elderly: empirical findings on age related changes and its contribution to satisfaction with life. AB - BACKGROUND: Positive psychology studies what is best in people. In their classification of strengths and virtues, Peterson and Seligman [11] assign humor to the virtue of transcendence. Thus far, there is no specific study that deals with age-related changes in humor (as a strength of character) across a lifespan and its relation to well-being in the elderly. PARTICIPANTS AND METHODS: A total of n=42,964 participants completed an online questionnaire on humor as a strength of character. Participants also completed the Satisfaction with Life Scale and the Orientations to Happiness Scale. RESULTS: In a cross-sectional design, the scores for humor decreased until the age of 50. Men between 51 and 62 years had higher scores; there was a trend for women older than 70 to score higher (but this was not statistically significant). Humor was robustly positively correlated with life satisfaction, as well with a pleasurable and an engaged life, but was lowest with a meaningful life. The oldest participants had the lowest (yet still meaningful) correlation coefficients (with the exception of a meaningful life). CONCLUSION: The study contributes to the understanding of humor across the lifespan and underlines the importance of studies among the elderly within a framework of positive psychology. PMID- 20012064 TI - [Humor and the brain: neurobiological aspects]. AB - In recent years, a number of studies have been published on cerebral activation induced by funny stimuli. This article provides an overview on the structures involved and findings concerning the neuropsychological faculties necessary for joke comprehension, e.g., working memory and mental flexibility. On the other hand, there are also many aspects of humor, like joke production, that are not neuropsychologically well studied; an overview of current knowledge will be given. In addition, there is little research on the effect of aging on the different aspects of cerebral humor processing. PMID- 20012065 TI - Sense of humor among the elderly: findings using the German version of the SHS. AB - Empirical studies on humor among the elderly are lacking. Based on the model of sense of humor by Paul McGhee, different age groups are compared (n=979) in the present study. Data suggest that playfulness and sense of humor are stable across different age groups (in a cross-sectional design) but that elderly participants were highest in positive mood. However, they also indicated that they laughed less and less easily than younger participants. Among the elderly, those who frequently (more than once a week) meet with friends and colleagues yielded higher expressions in different aspects of the sense of humor than those who meet friends and colleagues less often. Generally, humor was positively associated with personal and national well-being. However, among those participants older than 60 years of age, national well-being (i.e., satisfaction with the government, safety, or economics in the country) was negatively related to humor. Results are discussed together with a general outlook on the use of humor interventions for increasing the well-being and quality of life of elderly people. PMID- 20012066 TI - Humor as a character strength among the elderly: theoretical considerations. AB - Positive psychology is a scientific approach within psychology that focuses on research on what is best in people. Within this framework, humor is understood as a character strength. The article gives a brief overview on positive psychology and implications for aging (positive aging, successful aging) with an emphasis on the contribution of humor. Humor as a strength of character and its measurement by various approaches are discussed. It is argued that there is a lack of empirical data about humor in the elderly. Potential benefits of considering humor in research but also in practice are discussed. PMID- 20012067 TI - Forgotten cardiovascular diseases in Africa. AB - Much of the global burden of cardiovascular disease is now carried by low and middle income countries. Unfortunately, many of these regions are still grappling with poverty and infection-related cardiovascular diseases, such as endomyocardial fibrosis, tuberculous pericarditis and rheumatic heart disease. In addition, Africa has its unique diseases that occur more commonly in Africans as peripartum cardiomyopathy or, almost uniquely in Africa, as subvalvular aneurysm. We present our perspective on forgotten cardiovascular disease in Africa in the context of the epidemic of cardiovascular disease due to global changes in life style. PMID- 20012070 TI - Do we need femoral derotation osteotomy in DDH of early walking age group? A clinico-radiological correlation study. AB - INTRODUCTION: The occurrence of exaggerated femoral anteversion and the role of femoral derotation osteotomy in developmental dysplasia of hip, especially early walking age group are controversial. METHOD: We evaluated femoral anteversion, acetabular anteversion, acetabular index in 15 dislocated hips and 11 normal hips in cases of unilateral dislocation of hip in DDH of age group 12-48 months. We correlated this femoral anteversion with the intra operative "test of stability" which is described by Zadeh et al. We found that there was no statistically significant difference in femoral anteversion between dislocated and normal hips. In all the 15 cases we did open reduction by anterior approach (Somerville approach) and evaluated the position for maximum stability. In 3 cases we were unable to perform test of stability as they needed femoral shortening for reduction of joint. RESULTS: In the rest 12 hips, 10 were stable in flexion and abduction while 2 were stable in flexion. None of the hips required internal rotation for stability. Hence we did salters osteotomy in all the hips and femoral shortening through lateral approach in 3 cases. At a minimum follow up of 18 months all the hips were clinically stable and none of them dislocated till final follow up. The mean correction of acetabular index was 15.4 degrees and the outcome was excellent in 8 hips and good in 7 hips as per modified McKay's criteria. Hence we recommend that femoral derotation osteotomy is not needed in DDH of early walking age group. CONCLUSION: As the surgical treatment of DDH involves complex osteotomies around the hip and these surgeries have effect on long term outcome, MRI evaluation of femoral anteversion as a part of pre operative evaluation is advised. Also, as the intra operative evaluation needs enough clinical experience and it can not be performed in cases requiring femoral shortening for reduction, we consider pre operative evaluation of femoral anteversion by MRI as essential rather than adjunctive. PMID- 20012069 TI - Digit and hand replantation. AB - For the past 45 years, the advent of microsurgery has led to replantation of almost every amputated part such as distal phalanx, finger tip, etc. Replantation of digits and hand can restore not only circulation, but also function and cosmetic of the amputated part. The goals of replantation are to restore circulation and regain sufficient function and sensation of the amputated part. Strict selection criteria are necessary to optimize the functional result. The management of this type of injuries includes meticulous preoperative management, microsurgical experience and continuous postoperative care. Among various factors influencing the outcome, the most important are the type and the level of injury, ischemia time, history of diabetes, age, sex, and smoking history. During the replantation procedure, bone stabilization, tendon repair, arterial anastomoses, venous anastomoses, nerve coaptation, and skin coverage should be performed. All structures should be repaired primarily, unless a large nerve gap or a flexor tendon avulsion injury is present. Adequate postoperative evaluation is mandatory to avoid early or late complications. To improve functional results, many replantation patients may need further reconstructive surgery. PMID- 20012073 TI - Partial versus unrestricted weight bearing after an uncemented femoral stem in total hip arthroplasty: recommendation of a concise rehabilitation protocol from a systematic review of the literature. AB - The aim of this systematic review was to find evidence-based support in the literature to allow immediate unrestricted weight bearing after primary uncemented total hip arthroplasty (THA). Accelerated rehabilitation programs for THA are becoming increasingly popular to shorten hospital stay and to facilitate rapid restoration of function. The goals of these rehabilitation programs could be more easily achieved if immediate unrestricted weight bearing (UWB) could be allowed after a THA. So far, however, immediate weight bearing is frequently contraindicated in widely accepted protocols for uncemented THA due to fear for subsidence and absence of osseous integration of the femoral stem. Thus, frequently protected weight bearing and restricted activities are still advocated for at least 6 weeks after surgery. In addition, we analyzed the literature to come to a recommendation on gait pattern and walking aid. From a systematic search in several electronic databases 13 studies met the inclusion criteria. These studies were reviewed according to the Cochrane methodology. We found moderate to strong evidence that no adverse effects on subsidence and osseous integration of the femoral stem after uncemented THA occur after immediate UWB. Based on this literature review, we recommend early rehabilitation after uncemented THA with a reciprocally gait pattern using crutches, one cane for independency in ADL in case patients walk limp-free and walking without crutches as soon as possible. During the first weeks after surgery only stair climbing should be performed with protected weight bearing because of high torsion loads on the hip. PMID- 20012072 TI - Effect of partial medial meniscectomy on the proprioceptive function of the knee. AB - INTRODUCTION: The aim of this study was to assess the proprioception of the partial meniscectomized knee and to assess if there is an effect on the knee proprioception. MATERIALS AND METHODS: We performed a case-control study involving patients with meniscus tear at the posterior horn of medial meniscus. Group 1 composed of 19 patients, with an average age of 26.3 years, who were enrolled after sustaining an injury to the medial meniscus. Twenty healthy individuals, with an average age of 25.1, without history of knee injury, constituted Group 2. In order to document the proprioceptive capabilities of the knee, angle reproduction tests were performed. RESULTS: At a mean of 2 years after the surgery, there was no difference between the meniscectomized and healthy knee regarding the angle deviations in the lower flexion angles (15 degrees , 30 degrees , 45 degrees ). However, mean knee joint position sense (KJPS) at 60 degrees reproduction in Group 1 and 2 was 64.3 (SD 5.3) and 69.2 (SD 4.9), respectively. Concomitantly, mean KJPS at 75 degrees reproduction in Group 1 and 2 were 80.1 (SD 6.59) and 74.4 (SD 5.6), respectively. Paired t test showed a statistically significant difference between Group 1 and 2 when compared to control angle deviations at 60 degrees and 75 degrees reproductions (P < 0.05). CONCLUSION: KJPS of the patients in meniscectomized group was poorer than the patients in healthy knee group at 60 degrees and 75 degrees knee flexion degrees. Those results indicated that even a partial absence of menisci causes a deterioration of proprioceptive functions of the knee. PMID- 20012074 TI - Infected nonunions of diaphyseal fractures of the forearm. AB - BACKGROUND: There are no reported series that specifically deal with repair of infected nonunions of the diaphysis of the forearm bones. We sought to determine whether a standardized treatment protocol we have utilized for 15 patients from 1989 to 2005 results in a high union rate, resolution of infection, and a good functional outcome. METHODS: The study cohort included nine male and six female patients who presented to a University hospital setting with an infected nonunion of the diaphysis of the radius or ulna. Every patient had a minimum of 2-year follow-up. The average patient age was 45 years (range 17-79). Eight of the patients had fractures involving their dominant arm. Thirteen patients had initially fractured both the radius and ulna, but two of these patients had subsequently healed one of the bones. One patient had an isolated radius fractures, and one patient fractured the ulna alone. All patients underwent a protocol that combines aggressive surgical debridements as necessary, definitive fixation after 7-14 days, tricortical iliac crest bone grafting for segmental defects, leaving wounds open to heal by secondary intention, 6 weeks of culture specific intravenous antibiotics, and early active range of motion (ROM) exercises. We sought to report our success rate of nonunion repair, number of re interventions, complication rate, final ROM, and the ability to eradicate the infection using this treatment regimen. RESULTS: At most recent follow-up (average 5 years, range 2-15 years), all patients had united and resolved their infections. One case was considered a failure, although he did go on to unite a one-bone forearm and was free of infection at most recent follow-up. All but three patients, including the one failure, had at least 50 degrees of supination/pronation and 30-130 degrees of flexion/extension arc. Excluding the one failure that united his one-bone forearm at 46 months, the average time to union was 13.2 weeks (range 10-15 weeks). CONCLUSIONS: The results of this study indicate that our standard protocol for treatment of infected nonunion of the shafts of the radius and ulna is reliable at obtaining fracture union with a good functional result, while also resolving the infection. PMID- 20012068 TI - Astrocytes: biology and pathology. AB - Astrocytes are specialized glial cells that outnumber neurons by over fivefold. They contiguously tile the entire central nervous system (CNS) and exert many essential complex functions in the healthy CNS. Astrocytes respond to all forms of CNS insults through a process referred to as reactive astrogliosis, which has become a pathological hallmark of CNS structural lesions. Substantial progress has been made recently in determining functions and mechanisms of reactive astrogliosis and in identifying roles of astrocytes in CNS disorders and pathologies. A vast molecular arsenal at the disposal of reactive astrocytes is being defined. Transgenic mouse models are dissecting specific aspects of reactive astrocytosis and glial scar formation in vivo. Astrocyte involvement in specific clinicopathological entities is being defined. It is now clear that reactive astrogliosis is not a simple all-or-none phenomenon but is a finely gradated continuum of changes that occur in context-dependent manners regulated by specific signaling events. These changes range from reversible alterations in gene expression and cell hypertrophy with preservation of cellular domains and tissue structure, to long-lasting scar formation with rearrangement of tissue structure. Increasing evidence points towards the potential of reactive astrogliosis to play either primary or contributing roles in CNS disorders via loss of normal astrocyte functions or gain of abnormal effects. This article reviews (1) astrocyte functions in healthy CNS, (2) mechanisms and functions of reactive astrogliosis and glial scar formation, and (3) ways in which reactive astrocytes may cause or contribute to specific CNS disorders and lesions. PMID- 20012075 TI - Treatment of Laugier-Hunziker syndrome with the Q-switched alexandrite laser in 22 Chinese patients. AB - Laugier-Hunziker syndrome (LHS), a rare, acquired pigmentary disorder of the lips, oral mucosa, and fingers, is known to be an entirely benign disease with no systemic manifestations. In the past, the pigmentation has been treated efficiently in a few patients with the Q-switched neodymium: yttrium-aluminum garnet (Nd:YAG) laser and the Q-switched alexandrite laser (QSAL). In order to evaluate the efficacy and safety of QSAL on Chinese patients of LHS, we treated 22 patients with QSAL in the past 5 years. Treatments were delivered on a bimonthly or trimonthly basis until the abnormal pigmentation totally disappeared. Patients were evaluated at each visit for evidence of dyspigmentation, scarring, or other untoward effects from the laser treatment. Our 22 subjects consisted of 18 females and 4 males with a mean age of 42.4 years. After only one session of laser treatment, the clearing on the lips was as follow: 18 (81.8%) excellent, 2 (9.1%) good, 1 (4.5%) fair and 1 (4.5%) poor. Eighteen patients (81.8%) with LHS, who had achieved excellent clearing after only one session of laser treatment, did not receive further treatment. Among the left four patients, three patients (13.6%) achieved complete results after three laser treatments. Only one patient required six sessions to achieve complete clearance. No scarring was noted after any of the treatments. The appearance of pigmentation on mucous membranes in a middle-aged patient without a significant family history for skin disorders should prompt consideration for the possible diagnosis of LHS. Our study has also demonstrated QSAL to be highly effective and safe in the treatment of LHS. PMID- 20012076 TI - Single dose of preoperative analgesia with gabapentin (600 mg) is safe and effective in monitored anesthesia care for nasal surgery. AB - This study was aimed to compare the intraoperative sedative and perioperative analgesic drug requirements and the incidences of postoperative side effects on the patients who received preoperative gabapentin or placebo. Sixty patients undergoing nasal septal or nasal sinus surgery were included. The patients received either 600 mg gabapentin (Group G) or placebo (Group P) orally, 1 h before surgery. The scores for sedation and pain were recorded at 5, 15, 30, 45 and 60 min, intraoperatively and at 30 min, 1, 2, 4, 6, 9, 12, 16, 20, 24 h, postoperatively. Sedation was achieved with an IV bolus of propofol and continuous infusion of remifentanil. There were significant differences between gabapentin and placebo groups with regard to total consumptions of remifentanil (171.42 + or - 68 vs. 219.17 + or - 95 microg, respectively; P = 0.033) and propofol (59.45 + or - 36.08 vs. 104.14 + or - 54.98 mg, respectively; P = 0.001). Group G patients had significantly lower intraoperative VAS scores at all time points (P < 0.05). The anxiety score of Group G was better at all times (P < 0.05). All postoperative pain scores were lower in the Group G (P < 0.05). Time to first request for analgesic was 12.7 + or - 2.3 h in Group G, and 7.8 2.1 h in Group P (P < 0.0001). Total consumption of lornoxicam was lower in Group G (P < 0.004). We concluded that monitored anesthesia care combined with preoperative analgesia with a low dose of (600 mg) oral gabapentin is an efficient option with tolerable side effects for patients undergoing ear, nose and throat ambulatory surgery. PMID- 20012078 TI - A surgical view of the superior nasal turbinate: anatomical study. AB - Differences of the superior nasal turbinate (SNT), presence of the supreme nasal turbinate (SpNT) and measurements of opening sphenoid sinus (OSS) are consistent anatomical landmarks that allow for safe entrances, such as posterior ethmoidectomy and sphenoid sinusotomy. The purpose of study was to investigate the anatomical details of the SNT for approaching the OSS on 20 specimens of adult cadavers under an operating microscope. The SNT and SpNT were localized more perpendicular than parallel to their axes. The SpNT structure was observed in 12 specimens (60%) and it was classified into three types. Type A SpNT was shortest of all turbinates (58.3%). In types B and C, SpNT was equal or larger than the SNT. These types were seen in 41.7% of specimens. In 11 specimens, posterior ethmoidal cells opened to supreme nasal meatus. In 7 specimens, there was one opening to supreme nasal meatus, while 2 openings were detected in 12 specimens, and 3 openings were seen in 1 specimen. All these openings belonged to posterior ethmoidal cells. To determine the position of the OSS, distances between some anatomical points were measured. In cases where the SpNT is present or the SpNT is bigger than the SNT, it is certain that a different method will be applied during the procedure in the nasal cavity. The SNT and the SpNT may easily be injured by unrecognized dissection in types B and C, leading to the disruption of its olfactory neuroepithelium and possibly to postoperative hyposmia. PMID- 20012077 TI - Malignant transformation of an osteoblastoma of the mandible: case report and review of the literature. AB - Benign osteoblastoma is a rarely seen tumor of the facial bones. The authors present a case of a 30-year-old man with a tumor of the mandibular body and ramus. The histopathological diagnosis was one of osteoblastoma. Postoperative recurrence with soft tissue infiltration suggested an osteosarcoma radiologically, but the histological examination again revealed the presence of an osteoblastoma. A second recurrence occured in the pharyngo-glossal region and this time the tumor was histologically diagnosed as an osteoblastoma, but with foci of well-differentiated osteosarcoma. The patient was given a course of radiotherapy, but clinical and radiological examination 8 months later revealed lung metastases and chemotherapy was started. Unfortunately, the patient died months later. While osteoblastomas are rare, and their sarcomatous change even rarer, our experience with this case lead us to suggest that a therapeutic preventative approach, involving both chemotherapy and total excision of the tumor, is the regime to adopt with osteoblastomas which involve soft tissues and have radiological features suggesting malignancy. PMID- 20012080 TI - Distance and dynamics determination by W-band DEER and W-band ST-EPR. AB - To explore high-field EPR in biological applications we have compared measurements of dynamics with X-band (9 GHz) and W-band (94 GHz) saturation transfer EPR (ST-EPR) and distance determination by X and W-band DEER. A fourfold increase of sensitivity was observed for W-band ST-EPR compared with X-band. The distance measurements at both fields showed very good agreement in both the average distances and in the distance distributions. Multifrequency EPR thus provides an additional experimental dimension to facilitate extraction of distance populations. However, the expected orientational selectivity of W-band DEER to determine the relative orientation of spins has not been realized, most likely because of the large orientational disorder of spin labels on the protein surface. PMID- 20012081 TI - A systematic review on the effect of bevacizumab in exudative age-related macular degeneration. PMID- 20012079 TI - Serum protein profiling of smear-positive and smear-negative pulmonary tuberculosis using SELDI-TOF mass spectrometry. AB - The focus of this study was to detect novel sera biomarkers for smear-positive and smear-negative pulmonary tuberculosis and to establish respective diagnostic models using the surface-enhanced laser desorption ionization time-of-flight mass spectrometry (SELDI-TOF MS) technique. A total of 155 sera samples from smear positive pulmonary tuberculosis (SPPTB) and smear-negative pulmonary tuberculosis (SNPTB) patients and non-tuberculosis (non-TB) controls were analyzed with SELDI TOF MS. The study was divided into a preliminary training set and a blinded testing set. A classification tree of spectra derived from 31 SPPTB patients, 22 SNPTB patients, and 42 non-TB controls were used to develop an optimal classification tree that discriminated them respectively in the training set. Then, the validity of the classification tree was challenged with another independent blinded testing set, which included 20 SPPTB patients, 14 SNPTB patients, and 26 non-TB controls. SNPTB patients and non-TB controls also were analyzed alone using the same method. The optimal decision tree model with a panel of nine biomarkers with mass:charge ratios (m/z) of 4821.45, 3443.22, 9284.93, 4473.86, 4702.84, 3443.22, 5343.26, 3398.27, and 3193.61 determined in the training set could detect 93.55%, 95.46%, and 88.09% accuracy for classifying SPPTB patients, SNPTB patients, and non-TB controls specimens, respectively. Validation of an independent, blinded testing set gave an accuracy of 80.77% for controls, 75.00% for SPPTB, and 71.43% for SNPTB samples using the same classification tree. With the peaks displaying differences between SNPTB patients and non-TB controls, a simplified dendrogram (m/z 4821.45, 4792.74) demonstrated classification efficacy of 85.94% (sensitivity 86.36% and specificity 85.71%) for distinguishing SNPTB patients from non-TB controls. The independent blinded testing set containing 14 SNPTB patients and 26 non-TB controls gained an accuracy of 81.59% (sensitivity 78.57% and specificity 84.62%) for diagnosing SNPTB. Special proteins/peptides may change in SPPTB and SNPTB patients and those changes may be used to distinguish them with the proper discriminant analytical method and to pursue and identify some involved proteins underlying the biological process of tuberculosis. PMID- 20012082 TI - Parameters for a model of an oscillating neuronal network in the cochlear nucleus defined by genetic algorithms. AB - Chopper neurons in the cochlear nucleus are characterized by intrinsic oscillations with short average interspike intervals (ISIs) and relative level independence of their response (Pfeiffer, Exp Brain Res 1:220-235, 1966; Blackburn and Sachs, J Neurophysiol 62:1303-1329, 1989), properties which are unattained by models of single chopper neurons (e.g., Rothman and Manis, J Neurophysiol 89:3070-3082, 2003a). In order to achieve short ISIs, we optimized the time constants of Rothman and Manis single neuron model with genetic algorithms. Some parameters in the optimization, such as the temperature and the capacity of the cell, turned out to be crucial for the required acceleration of their response. In order to achieve the relative level independence, we have simulated an interconnected network consisting of Rothman and Manis neurons. The results indicate that by stabilization of intrinsic oscillations, it is possible to simulate the physiologically observed level independence of ISIs. As previously reviewed and demonstrated (Bahmer and Langner, Biol Cybern 95:371-379, 2006a), chopper neurons show a preference for ISIs which are multiples of 0.4 ms. It was also demonstrated that the network consisting of two optimized Rothman and Manis neurons which activate each other with synaptic delays of 0.4 ms shows a preference for ISIs of 0.8 ms. Oscillations with various multiples of 0.4 ms as ISIs may be derived from neurons in a more complex network that is activated by simultaneous input of an onset neuron and several auditory nerve fibers. PMID- 20012083 TI - Production and characterisation of the intergeneric hybrids between Dendranthema morifolium and Artemisia vulgaris exhibiting enhanced resistance to chrysanthemum aphid (Macrosiphoniella sanbourni). AB - Aphids represent the most destructive of chrysanthemum pests to cultivation. Reliable variety sources of resistance and control methods are limited, so development of highly resistant breeding lines is desirable. An intergeneric hybrid between Dendranthema morifolium (chrysanthemum) variety 'Zhongshanjingui' and Artemisia vulgaris (mugwort) 'Variegata' was attempted. Most of the hybrid embryos aborted at an early developmental stage. Embryo rescue allowed the generation of hybrid plants, whose hybridity was confirmed by a combination of morphological, cytological and GISH analysis. The hybrids were vigorous, flowered normally, and their flower and leaf shape resembled those of the chrysanthemum more than those of the mugwort parent. The hybrids showed much higher resistance to chrysanthemum aphid (Macrosiphoniella sanbourni) than maternal chrysanthemum by inoculation test. The leaves of the hybrid developed a higher density of trichomes and secretory glands compared to the maternal chrysanthemum. GC-MS analysis revealed that approximately 51% of the essential oil in the hybrid leaves were monoterpenoids and sesquiterpenoids, while the proportion in the chrysanthemum was approximately 37%, and in the mugwort was approximately 90%. It is inferred that higher aphid resistance in the hybrid mainly owed to the leaf micromorphology and bioactive essential oil content. PMID- 20012084 TI - A tomato enzyme synthesizes (+)-7-iso-jasmonoyl-L-isoleucine in wounded leaves. AB - Jasmonoyl-L-isoleucine (JA-Ile) is a key jasmonate signal that probably functions in all plant species. The JASMONATE RESISTANT 1 (JAR1) enzyme synthesizes JA-Ile in Arabidopsis [Arabidopsis thaliana (L.) Heynh.], but a similar enzyme from tomato [Solanum lycopersicum (L.)] was not previously described. Tomato SlJAR1 has 66% sequence identity with Arabidopsis JAR1 and the SlJAR1-GST fusion protein purified from Escherichia coli catalyzed the formation of JA-amino acid conjugates in vitro. Kinetic analysis showed the enzyme has a strong preference for Ile over Leu and Val and it was about 10-fold more active with (+)-7-iso-JA than with its epimer (-)-JA. Leaf wounding rapidly increased JA-Ile 50-fold to about 450 pmol g(-1) FW at 30 min after wounding, while conjugates with Leu, Phe, Val and Met were only marginally increased or not detected. Nearly all of the endogenous JA-Ile was the bioactive epimer (+)-7-iso-JA-Ile and there was no evidence for its conversion to (-)-JA-Ile up to 6 h after wounding. A transgenic RNAi approach was used to suppress SlJAR1 transcript that reduced JA-Ile accumulation by 50-75%, suggesting that other JA conjugating enzymes may be present. These results show that SlJAR1 synthesizes the bioactive conjugate (+)-7 iso-JA-Ile and this is the predominant isomer accumulated in wounded tomato leaves. PMID- 20012086 TI - Down-regulation of four putative arabinoxylan feruloyl transferase genes from family PF02458 reduces ester-linked ferulate content in rice cell walls. AB - Industrial processes to produce ethanol from lignocellulosic materials are available, but improved efficiency is necessary to make them economically viable. One of the limitations for lignocellulosic conversion to ethanol is the inaccessibility of the cellulose and hemicelluloses within the tight cell wall matrix. Ferulates (FA) can cross-link different arabinoxylan molecules in the cell wall of grasses via diferulate and oligoferulate bridges. This complex cross linking is thought to be a key factor in limiting the biodegradability of grass cell walls and, therefore, the reduction in FA is an attractive target to improve enzyme accessibility to cellulose and hemicelluloses. Unfortunately, our knowledge of the genes responsible for the incorporation of FA to the cell wall is limited. A bioinformatics prediction based on the gene similarities and higher transcript abundance in grasses relative to dicot species suggested that genes from the pfam family PF02458 may act as arabinoxylan feruloyl transferases. We show here that the FA content in the cell walls and the transcript levels of rice genes Os05g08640, Os06g39470, Os01g09010 and Os06g39390, are both higher in the stems than in the leaves. In addition, an RNA interference (RNAi) construct that simultaneously down-regulates transcript levels of these four genes is associated with a significant reduction in FA of the cell walls from the leaves of the transgenic plants relative to the control (19% reduction, P < 0.0001). Therefore, our experimental results in rice support the bioinformatics prediction that members of family PF02458 are involved in the incorporation of FA into the cell wall in grasses. PMID- 20012085 TI - miR398 and miR408 are up-regulated in response to water deficit in Medicago truncatula. AB - Plant microRNAs have been implicated in various abiotic stress responses. We identified several conserved microRNAs that showed differential expression in Medicago truncatula plants subjected to water deficit: miR169 is down-regulated only in the roots and miR398a/b and miR408 are strongly up-regulated in both shoots and roots. Down-regulation of miR169 in the roots did not correlate with accumulation of its target MtHAP2-1 transcripts, suggesting that its regulation may not occur at the mRNA level or may depend on other regulatory mechanisms, which do not involve this miRNA, in water-deficit conditions. The up-regulation of miR398a/b and miR408 and the clear down-regulation of their respective target genes, which encode the copper proteins COX5b (subunit 5b of mitochondrial cytochrome c oxidase) and plantacyanin, highlight the involvement of these miRNAs in response to water deprivation in M. truncatula. Also, miR398 up-regulation is inversely correlated with the down-regulation of copper superoxide dismutase, CSD1, during water deficit. The regulation of genes encoding copper proteins by miR398a/b and miR408 suggests a link between copper homeostasis and M. truncatula adaptation to progressive water deficit. PMID- 20012087 TI - Genes involved in ethylene and gibberellins metabolism are required for endosperm limited germination of Sisymbrium officinale L. seeds: germination in Sisymbrium officinale L. seeds. AB - The rupture of the seed coat and that of the endosperm were found to be two sequential events in the germination of Sisymbrium officinale L. seeds, and radicle protrusion did not occur exactly in the micropylar area but in the neighboring zone. The germination patterns were similar both in the presence of gibberellins (GA(4+7)) and in presence of ethrel. The analysis of genes involved in GAs synthesis and breakdown demonstrated that (1) SoGA2ox6 expression peaked just prior to radicle protrusion (20-22 h), while SoGA3ox2 and SoGA20ox2 expression was high at early imbibition (6 h) diminishing sharply thereafter; (2) the accumulation of SoGA20ox2 transcript was strongly inhibited by paclobutrazol (PB) as well as by inhibitors of ET synthesis and signaling (IESS) early after imbibition (6 h), while SoGA3ox2 and SoGA2ox6 expression was slowly depressed as germination progressed; (3) ethrel and GA(4+7) positively or negatively affected expression of SoGA3ox2, SoGA20ox2, and SoGA2ox6, depending on the germination period studied. Regarding genes involved in ET synthesis, our results showed that SoACS7 was expressed, just prior to radicle emergence while SoACO2 expression slowly increased as germination progressed. Both genes were strongly inhibited by PB but were almost unaffected by externally added ethrel or GA(4+7). These results suggest that GAs are more important than ET during the early stages of imbibition, while ET is more important at the late phases of germination of S. officinale L. seeds. PMID- 20012088 TI - Light exaggerates apical hook curvature through phytochrome actions in tomato seedlings. AB - Contrary to the established notion that the apical hook of dark-grown dicotyledonous seedlings opens in response to light, we found in tomato (Solanum lycopersicum L.) that the apical hook curvature is exaggerated by light. Experiments with several tomato cultivars and phytochrome mutants, irradiated with red and far-red light either as a brief pulse (Rp, FRp) or continuously (Rc, FRc), revealed: the hook-exaggeration response is maximal at the emergence of the hypocotyl from the seed; the effect of Rp is FRp-reversible; fluence-response curves to a single Rp or FRp show an involvement of low and very low fluence responses (LFR, VLFR); the effect of Rc is fluence-rate dependent, but that of FRc is not; the phyA mutant (phyA hp-1) failed to respond to an Rp of less than 10(-2) micromol m(-2) and to an FRp of all fluences tested as well as to FRc, thus indicating that the hook-exaggeration response involves phyA-mediated VLFR. The Rp fluence-response curve with the same mutant also confirmed the presence of an LFR mediated by phytochrome(s) other than phyA, although the phyB1 mutant (phyB1 hp-1) still showed full response probably due to other redundant phytochrome species (e.g., phyB2). Simulation experiments led to the possible significance of hook exaggeration in the field that the photoresponse may facilitate the release of seed coat when seeds germinate at some range of depth in soil. It was also observed that seed coat and/or endosperm are essential to the hook exaggeration. PMID- 20012089 TI - Comparison of histopathological and gene expression-based typing of cancer of unknown primary. AB - For the clinical diagnosis and appropriate therapy of patients with cancer of unknown primary (CUP), biopsies of the tumor metastases are generally examined histopathologically and by immunohistochemistry (IHC). Gene expression profiling (GEP) is a new diagnostic technique that might further contribute to tumor specification. Biopsies of 43 CUP cases underwent a retrospective central histopathological and immunohistochemical review and centrally performed GEP using CupPrint(TM). Two samples could not be evaluated by IHC due to small biopsy size or loss of immunoreactivity. One of these and 17 other samples were not suited for GEP due to RNA degradation. In 13 out of the remaining 24 cases (54%), the same primary was proposed by IHC and GEP and was supported by the clinical findings, furthermore leading to the doubtless identification of the primary in four out of these. In seven cases, there was discordance between IHC and GEP, with the clinical picture being more in line with IHC in three and with GEP in four cases. Four cases had to remain undecided because the primary tumors suggested by IHC and GEP were not supported. In conclusion, overlap between IHC and GEP results and the clinical presentation was noted in the majority of those true CUP cases that were evaluable with both techniques. Therefore, GEP can be a complementary diagnostic technique assisting immunohistochemical profiling of cancer biopsies with unknown primary. PMID- 20012090 TI - Thoracic vasculitis presenting as surgical problems. AB - We present four patients with vasculitis manifesting with unusual clinical or pathological features, generating surgical problems. Two cases presented with pulmonary hypertension, with investigations and radiological evidence prompting clinical suspicion of pulmonary thrombo-embolic disease. First case, with an antecedant history of Wegener's granulomatosis (WG), demonstrated following "embolectomy", WG involving the large pulmonary elastic arteries. The second case of inoperable "pulmonary thrombo-embolic disease" was subsequently found at limited post mortem to have giant cell arteritis, which affected widespread small peripheral pulmonary arterial vessels. The other two cases were of aortitis occurring in the background of immune-mediated disease, which had been treated with aggressive immunosuppression regimens. The first of these was a case of Cogan's syndrome complicated by descending aortitis, a rarely reported phenomenon, with co-existent acute endocarditis of the aortic valve leaflets. Most cases of endocarditis in this context occur secondary to and in continuity with ascending aortitis. That this case, and a case of ascending aortitis occurring in the context of relapsing polychondritis occurred in the face of aggressive immunosuppression with an apparent clinical response, underscores the need to not accept a clinical picture at face value. This has implications for clinical management, particularly in the follow-up of surgical prosthetic devices such as grafts which may be used in these cases. All four cases emphasise the continued importance of histology and the post-mortem examination in elucidating previously undetected or unsuspected disease. PMID- 20012093 TI - MnSOD Val16Ala polymorphism and prostate cancer susceptibility: a meta-analysis involving 8,962 subjects. AB - PURPOSE: Published data on the association between manganese superoxide dismutase (MnSOD) Val(16)Ala polymorphism and prostate cancer (PCA) risk are inconclusive. To derive a more precise estimate of the association between them, a meta analysis was performed. METHODS: PubMed and Embase were searched. All eligible studies were retrieved. The pooled odds ratio (OR) with 95% confidence interval (CI) for PCA risk associated with Val/Ala versus Val/Val, Ala/Ala versus Val/Val, dominant model (Ala/Ala + Val/Ala vs. Val/Val), and recessive model (Ala/Ala vs. Val/Ala + Val/Val) were estimated, respectively. RESULTS: A total of 12 studies including 8,962 subjects were involved in this meta-analysis. Overall, the meta analysis indicated that significantly elevated cancer risk was associated with Ala variant genotype when all the eligible studies were pooled into the meta analysis (for Val/Ala vs. Val/Val: OR = 1.11, 95% CI = 1.00-1.24; for Ala/Ala vs. Val/Val: OR = 1.22, 95% CI = 1.00-1.49; for dominant model: OR = 1.14, 95% CI = 1.03-1.26). In the subgroup analysis by ethnicity, statistically significant increased risks were found among Caucasians with Ala allele (for Val/Ala vs. Val/Val: OR = 1.12, 95% CI = 1.00-1.25; for dominant model: OR = 1.14, 95% CI = 1.02-1.26). However, no significant associations were found in Africans. CONCLUSIONS: This meta-analysis suggests that the Ala allele of the MnSOD gene was a low-penetrance susceptible gene in PCA development, especially in Caucasians. PMID- 20012091 TI - Immunotherapeutic approaches for Alzheimer's disease in transgenic mouse models. AB - Alzheimer's disease (AD) is a member of a category of neurodegenerative diseases characterized by the conformational change of a normal protein into a pathological conformer with a high beta-sheet content that renders it resistant to degradation and neurotoxic. In the case of AD the normal soluble amyloid beta (sAbeta) peptide is converted into oligomeric/fibrillar Abeta. The oligomeric forms of Abeta are thought to be the most toxic, while fibrillar Abeta becomes deposited as amyloid plaques and congophilic angiopathy, which both serve as neuropathological markers of the disease. In addition, the accumulation of abnormally phosphorylated tau as soluble toxic oligomers and as neurofibrillary tangles is an essential part of the pathology. Many therapeutic interventions are under investigation to prevent and treat AD. The testing of these diverse approaches to ameliorate AD pathology has been made possible by the existence of numerous transgenic mouse models which each mirror different aspects of AD pathology. Perhaps the most exciting of these approaches is immunomodulation. Vaccination is currently being tried for a range of age associated CNS disorders with great success being reported in many transgenic mouse models. However, there is a discrepancy between these results and current human clinical trials which highlights the limitations of current models and also uncertainties in our understanding of the underlying pathogenesis of AD. No current AD Tg mouse model exactly reflects all aspects of the human disease. Since the underlying etiology of sporadic AD is unknown, the process of creating better Tg models is in constant evolution. This is an essential goal since it will be necessary to develop therapeutic approaches which will be highly effective in humans. PMID- 20012094 TI - Gene-environment interactions associated with CYP1A1 MspI and GST polymorphisms and the risk of upper aerodigestive tract cancers in an Indian population. AB - PURPOSE: Genetic risk to tobacco related cancers are associated with polymorphisms in CYP1A1 and GST, which are involved in the metabolic activation and detoxification of carcinogens. The genetic variations in these drug metabolizing enzymes may alter the susceptibility to UADT cancers triggered by environmental exposures. The hospital-based case-control study evaluated the impact of combined CYP1A1 MspI and GST (M1 & T1) polymorphisms among the individuals exposed to environmental risk factors as modulators in the risk of UADT cancers in Tamilians, a population of south India. METHODS: The unrelated histopathologically confirmed 408 cases and 220 population-based controls matched by age and gender were genotyped for CYP1A1 MspI, GSTM1 and GSTT1 polymorphisms using PCR based methods. To investigate the potential gene-environment interactions, analyses were carried out stratifying by smoking and tobacco chewing status using SPSS software. RESULTS: The combination of genes and environment interactions by stratified analyses revealed significant interactions among the habitual tobacco smokers (CYP1A1 MspI & GSTM1 null: OR 14.06; 95% CI 3.90-50.68, CYP1A1 MspI & GSTT1 null: OR 33.28; 95% CI 4.24-261.19) and tobacco chewers (CYP1A1 MspI & GSTM1 null: OR 20.51; 95% CI 6.77-62.13, CYP1A1 MspI & GSTT1 null: OR 79.35; 95% CI 10.40-605.55) on the multiplicative scale. CONCLUSION: Our findings have indicated that the individuals polymorphic for CYP1A1 MspI either with GSTM1 null or with GSTT1 null genotypes revealed an increased risk for UADT cancers than that ascribed to a single susceptible gene among the tobacco users in the population [single gene risk among smokers and chewers, respectively, for CYP1A1 MspI (OR 6.43; 95% CI 3.69-11.21); (OR 10.24; 95% CI 5.95-17.60), GSTM1*0 (OR 3.77; 95% CI 1.94-7.37); (OR 7.97 95% CI 4.10 15.76) and GSTT1*0 (OR 6.95 95% CI 2.88-16.77); (OR 25.83 95% CI 7.78-85.76). PMID- 20012095 TI - Evaluation of the vectorial capacity of Rhipicephalus sanguineus (Acari: Ixodidae) in the transmission of canine visceral leishmaniasis. AB - The vectorial capacity of Rhipicephalus sanguineus in the transmission of canine visceral leishmaniasis has been evaluated through a laboratory-controlled experiment. One healthy Leishmania-free dog and two dogs naturally infected with Leishmania were infested with R. sanguineus in various stages of development. Engorged larvae, unfed nymphs, engorged nymphs, unfed adults, engorged female adults and fed male adults were collected from the experimental animals and examined for Leishmania infection by optical microscopy, polymerase chain reaction (PCR) and parasite culture. Leishmania forms were not detected in any of the 433 smears prepared from engorged colonies nor in any of the 118 smears prepared from unfed colonies. However, one flagellate structure was identified in one of the smears. All pools of R. sanguineus that had fed on the infected dogs tested PCR-positive for Leishmania DNA, with the single exception of the pool of engorged larvae. In contrast, all pools of ticks that had fed on the Leishmania free dog were PCR-negative. Leishmania growth was not observed in any of the tick colonies following incubation on culture medium. Considering that no Leishmania forms were identified in any of the meticulously analysed smears derived from engorged colonies of R. sanguineus, it appears somewhat unlikely that the maintenance and multiplication of Leishmania occurs within the tick. PMID- 20012092 TI - Understanding the molecular basis of Alzheimer's disease using a Caenorhabditis elegans model system. AB - Alzheimer's disease (AD) is the major cause of dementia in the United States. At the cellular level, the brains of AD patients are characterized by extracellular dense plaques and intracellular neurofibrillary tangles whose major components are the beta-amyloid peptide and tau, respectively. The beta-amyloid peptide is a cleavage product of the amyloid precursor protein (APP); mutations in APP have been correlated with a small number of cases of familial Alzheimer's disease. APP is the canonical member of the APP family, whose functions remain unclear. The nematode Caenorhabditis elegans, one of the premier genetic workhorses, is being used in a variety of ways to address the functions of APP and determine how the beta-amyloid peptide and tau can induce toxicity. First, the function of the C. elegans APP-related gene, apl-1, is being examined. Although different organisms may use APP and related proteins, such as APL-1, in different functional contexts, the pathways in which they function and the molecules with which they interact are usually conserved. Second, components of the gamma-secretase complex and their respective functions are being revealed through genetic analyses in C. elegans. Third, to address questions of toxicity, onset of degeneration, and protective mechanisms, different human beta-amyloid peptide and tau variants are being introduced into C. elegans and the resultant transgenic lines examined. Here, we summarize how a simple system such as C. elegans can be used as a model to understand APP function and suppression of beta-amyloid peptide and tau toxicity in higher organisms. PMID- 20012096 TI - Sequence diversity of internal transcribed spacer-1 (ITS-1) region of Eimeria infecting chicken and its relevance in species identification from Indian field samples. AB - Conventional method of species identification in Eimeria employs phenotypic characters of the oocysts and the site of infection in the chicken intestine, which are subjective analyses. PCR-based identification of Eimeria spp. is known to be specific and sensitive. We used internal transcribed spacer 1 (ITS-1)-based nested PCR to follow the distribution of Eimeria spp. in the field, which may be of significant value in the management of coccidiosis in chickens. In the present study, intestinal samples of chicks from commercial poultry farms, in India, suspected of having contracted Eimeria infections were analyzed using ITS-1 PCR. The PCR-amplified ITS-1 regions were also sequenced from these samples. Of 26 field samples analyzed, 19 showed the presence of multiple infections of Eimeria spp. Incidence of Eimeria tenella (80%) was found to be highest in these samples followed by Eimeria mitis (53%), Eimeria acervulina (42%), Eimeria brunetti, and Eimeria maxima (23%). Incidence of Eimeria necatrix was found to be the lowest (15%) in the samples analyzed, while none of the samples analyzed showed the presence of ITS-1 sequence from Eimeria praecox. The ITS-1 sequences amplified from Eimeria spp. in the present study showed few variations from the ITS sequences available in the GenBank database. Further studies will be required to determine whether these differences are unique to geographical locations. PMID- 20012098 TI - Species-soil associations, disturbance, and nutrient cycling in an Australian tropical rainforest. AB - Resource availability and disturbance are important factors that shape the composition, structure, and functioning of ecosystems. We investigated the effects of soil fertility and disturbance on plant-soil interactions and nutrient cycling in a diverse tropical rainforest. Our goal was to determine how common soil specialisation is among species and how plant-soil interactions affect ecosystem functioning in the presence of disturbance. Most species (59%) showed significant fidelity to either fertile (basalt) or infertile (schist) soils. Obligate schist specialists (six species) contributed 39 and 37% to total stand level basal area and aboveground net primary productivity, respectively. High nutrient use efficiency of schist specialists reduced the rates of within-stand nutrient cycling through the production of nutrient-poor plant tissues and litter. Although forests on schist soils had higher basal area and similar rates of productivity to forests on basalt, uptake of Mg, K, P, and N were markedly less on schist than on basalt, particularly after a cyclone disturbance. Stands on schist soils were also less affected by the cyclone and, as a result, contributed less (ca. 50%) Mg, K, P, and N inputs to the forest floor (via litterfall) than stands on basalt soils. System "openness" (i.e. the risk of nutrient loss) from cyclone-affected basalt forests was minimised by high rates of uptake following disturbance and large effective cation exchange capacities of soils. Soil-plant-disturbance interactions are likely to engender different fitness-enhancing strategies on fertile and infertile soils, possibly leading to the development and/or maintenance of diversity in rainforests. PMID- 20012097 TI - Ecosystem-phase interactions: aquatic eutrophication decreases terrestrial plant diversity in California vernal pools. AB - Eutrophication has long been known to negatively affect aquatic and terrestrial ecosystems worldwide. In freshwater ecosystems, excessive nutrient input results in a shift from vascular plant dominance to algal dominance, while the nutrient species richness relationship is found to be unimodal. Eutrophication studies are usually conducted in continuously aquatic or terrestrial habitats, but it is unclear how these patterns may be altered by temporal heterogeneity driven by precipitation and temperature variation. The California vernal pool (CVP) ecosystem consists of three distinct phases (aquatic, terrestrial, and dry) caused by variation in climatic conditions. The purpose of this study was to test the hypothesis that resource addition during the aquatic phase results in increased algal abundance, which reduces vascular plant cover and richness of the terrestrial phase upon desiccation. We used mesocosms layered with CVP soil, in which treatments consisted of five levels of nitrogen and phosphorous added every 2 weeks. Resource addition increased available phosphorus levels and algae cover during the aquatic phase. Increased algal crusts resulted in decreased vascular plant percent cover and species richness. Few significant patterns were observed with individual plant species and total biomass. The phosphorus-plant richness relationship was not significant, but species composition was significantly different among the low and high treatment comparisons. These results highlight a neglected effect of eutrophication in seasonal habitats. Interactions among ecosystem phases clearly require more attention empirically and theoretically. Management and restoration of temporally heterogeneous habitat, such as the endemic-rich CVP, need to consider the extensive effects of increased nutrient input. PMID- 20012099 TI - Plant-soil feedback of native and range-expanding plant species is insensitive to temperature. AB - Temperature change affects many aboveground and belowground ecosystem processes. Here we investigate the effect of a 5 degrees C temperature increase on plant soil feedback. We compare plant species from a temperate climate region with immigrant plants that originate from warmer regions and have recently shifted their range polewards. We tested whether the magnitude of plant-soil feedback is affected by ambient temperature and whether the effect of temperature differs between these groups of plant species. Six European/Eurasian plant species that recently colonized the Netherlands (non-natives), and six related species (natives) from the Netherlands were selected. Plant-soil feedback of these species was determined by comparing performance in conspecific and heterospecific soils. In order to test the effect of temperature on these plant-soil feedback interactions, the experiments were performed at two greenhouse temperatures of 20/15 degrees C and 25/20 degrees C, respectively. Inoculation with unconditioned soil had the same effect on natives and non-natives. However, the effect of conspecific conditioned soil was negative compared to heterospecific soil for natives, but was positive for non-natives. In both cases, plant-soil interactions were not affected by temperature. Therefore, we conclude that the temperature component of climate change does not affect the direction, or strength of plant soil feedback, neither for native nor for non-native plant species. However, as the non-natives have a more positive soil feedback than natives, climate warming may introduce new plant species in temperate regions that have less soil-borne control of abundance. PMID- 20012100 TI - Levels of antioxidants in rural and urban birds and their consequences. AB - Numerous animals have successfully invaded urban habitats, although the factors associated with invasion success remain poorly understood. Urban areas are characterized by warmer microclimates, higher levels of primary productivity, longer breeding seasons and higher levels of pollutants. All these factors should cause oxidative stress, favoring invasion by species that have access to high levels of antioxidants. We analyzed concentrations of two categories of dietary, fat-soluble antioxidants (total carotenoids, total vitamin E) in the liver, the main storage organ in birds. Individuals killed by cats had lower levels of vitamin E than individuals that died for other reasons, showing natural selection on stored antioxidants. Bird species that had successfully colonized urban areas had significantly higher levels of vitamin E and total carotenoids than species that did not succeed, and rural populations had higher concentrations of vitamin E and total carotenoids than urban populations of the same species. Interspecific differences in concentrations of fat-soluble antioxidants, and differences between rural and urban populations of the same species, were accounted for by diet, but also by time since urbanization and number of generations since urbanization. These findings suggest that antioxidants, and by implication the ability to cope with oxidative stress, have contributed to successful invasion of urban areas by birds, and that the concentration of these antioxidants has changed in response to the urban environment. PMID- 20012101 TI - Growth and chemical defense in willow seedlings: trade-offs are transient. AB - Many studies have failed to detect costs of defense and some have even found a positive correlation between growth and the concentrations of chemical defenses. These studies contradict the theoretical assumption that anti-herbivore defenses are costly-produced at the expense of growth and/or reproduction. Costs, however, may be transient and therefore difficult to detect. Here we tested the hypothesis that costs of defense would be pronounced early in development when root growth is prioritized (high percent root allocation), but not later in development. To test this hypothesis, we grew F(2) hybrid willow seedlings from five different families, and harvested cohorts of even-aged seedlings after 6, 7, 8 and 9 weeks of growth. Seedlings were divided into root and shoot tissue and shoots were analyzed for phenolics (condensed tannins and phenolic glycosides). We found evidence for transient costs of defense. The concentrations of phenolics were negatively correlated with total biomass, shoot biomass, and the proportion of biomass allocated to roots in week 6. After week 6, however, the concentrations of phenolics were positively correlated with shoot biomass and total biomass, while phenolics were uncorrelated with the proportion of biomass allocated to roots. These results, the first ever, to our knowledge, with woody plants, suggest that costs of defense were transient; specifically, costs were found in early development, when root establishment was a priority. Our findings suggest that studies should focus more on trade-offs early in plant development. PMID- 20012104 TI - Metabolic syndrome risk factors and estimated glomerular filtration rate among children and adolescents. AB - The aim of this study was to seek the possible relationship between estimated glomerular filtration rate (e-GFR) and anthropometric indexes, lipids, insulin sensitivity, and metabolic syndrome risk factors among healthy children and adolescents. Sufficient evidence suggest that obesity is related with a novel form of glomerulopathy named obesity-related glomerulopathy (ORG) among adults, children, and adolescents. Glomerular filtration rate was estimated from serum creatinine in 166 healthy children and adolescents [79 males, 87 females; age 10.6 +/- 3.3 (3-18) years]. Anthropometric indexes and systolic and diastolic blood pressure were measured. Fasting insulin, glucose, creatinine, uric acid, total cholesterol, high-density lipoprotein (HDL)-cholesterol, low-density lipoprotein (LDL)-cholesterol, and triglycerides were estimated. Insulin sensitivity was estimated from known formulas. The presence of certain metabolic syndrome risk factors was checked among the studied population. Boys showed higher e-GFR rates than girls (f = 8.49, p = 0.004). We found a strong positive correlation between e-GFR and body weight (r = 0.415), body mass index (BMI) (r = 0.28), waist circumference (r = 0.419), hip circumference (r = 0.364), birth weight (r = 0.164), systolic blood pressure (SBP) (r = 0.305), and mean arterial pressure (MAP) (r = 0.207). A negative correlation was found between e-GFR and fasting glucose (r = -0.19), total cholesterol (r = -0.27) and LDL-cholesterol (r = -0.26). Clustering of metabolic syndrome risk factors among certain individuals was correlated with higher e-GFR rates (f = 3.606, p = 0.007). The results of this study suggest that gender, anthropometric indexes, and SBP are strong positive determinants of e-GFR among children and adolescents. Waist circumference is the most powerful determinant of e-GFR. Fasting glucose and lipid abnormalities are negative determinants of e-GFR among the studied population. Clustering of metabolic syndrome risk factors is coupled with higher e-GFR rates. PMID- 20012106 TI - The 5-HT4 receptor agonist mosapride attenuates NSAID-induced gastric mucosal damage. AB - BACKGROUND: The cholinergic anti-inflammatory pathway is a novel physiological mechanism found at various locations in the body where the nicotinic regulation of inflammatory cells through the autonomic nervous system is involved. In this study, we tested the hypothesis that cholinergic nerve stimulation by a 5-HT(4) agonist may modulate the progression of gastric mucosal ulcers induced by nonsteroidal anti-inflammatory drugs (NSAIDs). METHODS: Acute gastric ulcers were induced in rats by the oral administration of indomethacin. RESULTS: Gastric damage analysis indicated that pretreatment with mosapride, a selective 5-HT(4) agonist, at 0.25, 0.5, and 0.75 mg/kg, inhibited the mucosal damage induced by indomethacin. In gastric emptying analysis, an evacuation effect was observed in the 3.0 mg/kg mosapride pretreatment group, but this effect was not observed in the lower dose (0.5 mg/kg) group. The antiulcerogenic activity of mosapride treatment (at 0.5 mg/kg) was blocked by a 5-HT(4)-specific antagonist, GR113808 (1 mg/kg, i.v.). Additionally, we demonstrated that methyllycaconitine (0.29 and 0.87 mg/kg i.p.), a selective inhibitor of alpha7 nicotinic acetylcholine (ACh) receptors (alpha7nAChRs), ablated the antiulcerogenic action of mosapride. CONCLUSIONS: These results suggest that the mucosal protective action of mosapride may be mediated by an action on immune cells through the acceleration of ACh release from parasympathetic nerves via the activation of 5-HT(4) receptors, followed by activation of the nicotinic anti-inflammatory system. It appears that the alpha7nAChR may be involved in the antiulcerogenic action of mosapride. PMID- 20012107 TI - The complement component C3a fragment is a potential biomarker for hepatitis C virus-related hepatocellular carcinoma. AB - BACKGROUND: Hepatocellular carcinoma (HCC) has a high mortality rate, and early detection of HCC improves patient survival. However, the molecular diagnostic markers for early HCC have not been fully elucidated. The aim of this study was to identify novel diagnostic markers for HCC. METHODS: Serum protein profiles of 45 hepatitis C virus infection (HCV)-related HCC patients (HCV-HCC) were compared to 42 HCV-related chronic liver disease patients without HCC (HCV-CLD) and 21 healthy volunteers using the ProteinChip SELDI system. One of the identified proteins was evaluated as a diagnostic marker for HCC in patients with HCV. RESULTS: Five protein peaks (4067, 4470, 7564, 7929, and 8130 m/z) had p-values less than 1 x 10(-7) and were significantly increased in the sera of HCV-HCC patients compared to HCV-CLD patients and healthy volunteers. Among these proteins, an 8130 m/z peak was the most differentially expressed and identified as the complement component 3a (C3a) fragment. For HCV-HCC and HCV-CLD, the relative intensity of this C3a fragment had the best area under the ROC curve [0.70], followed by des-gamma-carboxy prothrombin (DCP) [0.68], lectin-bound alpha fetoprotein (AFP-L3) [0.58] and AFP [0.53] for HCC. A combined analysis of the C3a fragment, AFP and DCP led to a 98% positive identification rate. In addition, the measurable C3a fragment in some HCC patients was not only significantly higher in the year of HCC onset compared to the pre-onset year, but also decreased after treatment. CONCLUSIONS: The 8130 m/z C3a fragment is a potential marker for the early detection of HCV-related HCC. PMID- 20012108 TI - Fungal root endophytes of the carnivorous plant Drosera rotundifolia. AB - As carnivorous plants acquire substantial amounts of nutrients from the digestion of their prey, mycorrhizal associations are considered to be redundant; however, fungal root endophytes have rarely been examined. As endophytic fungi can have profound impacts on plant communities, we aim to determine the extent of fungal root colonisation of the carnivorous plant Drosera rotundifolia at two points in the growing season (spring and summer). We have used a culture-dependent method to isolate fungal endophytes and diagnostic polymerase chain reaction methods to determine arbuscular mycorrhizal fungi colonisation. All of the roots sampled contained culturable fungal root endophytes; additionally, we have provided molecular evidence that they also host arbuscular mycorrhizal fungi. Colonisation showed seasonal differences: Roots in the spring were colonised by Articulospora tetracladia, two isolates of uncultured ectomycorrhizal fungi, an unidentified species of fungal endophyte and Trichoderma viride, which was present in every plant sampled. In contrast, roots in the summer were colonised by Alatospora acuminata, an uncultured ectomycorrhizal fungus, Penicillium pinophilum and an uncultured fungal clone. Although the functional roles of fungal endophytes of D. rotundifolia are unknown, colonisation may (a) confer abiotic stress tolerance, (b) facilitate the acquisition of scarce nutrients particularly at the beginning of the growing season or (c) play a role in nutrient signalling between root and shoot. PMID- 20012109 TI - Neuropathological heterogeneity in frontotemporal lobar degeneration with TDP-43 proteinopathy: a quantitative study of 94 cases using principal components analysis. AB - Studies suggest that frontotemporal lobar degeneration with transactive response DNA-binding protein of 43 kDa (TDP-43) proteinopathy (FTLD-TDP) is heterogeneous with division into four or five subtypes. To determine the degree of heterogeneity and the validity of the subtypes, we studied neuropathological variation within the frontal and temporal lobes of 94 cases of FTLD-TDP using quantitative estimates of density and principal components analysis (PCA). A PCA based on the density of TDP-43 immunoreactive neuronal cytoplasmic inclusions, oligodendroglial inclusions, neuronal intranuclear inclusions, and dystrophic neurites, surviving neurons, enlarged neurons, and vacuolation suggested that cases were not segregated into distinct subtypes. Variation in the density of the vacuoles was the greatest source of variation between cases. A PCA based on TDP 43 pathology alone suggested that cases of FTLD-TDP with progranulin (GRN) mutation segregated to some degree. The pathological phenotype of all four subtypes overlapped but subtypes 1 and 4 were the most distinctive. Cases with coexisting motor neuron disease (MND) or hippocampal sclerosis (HS) also appeared to segregate to some extent. We suggest: (1) pathological variation in FTLD-TDP is best described as a 'continuum' without clearly distinct subtypes, (2) vacuolation was the single greatest source of variation and reflects the 'stage' of the disease, and (3) within the FTLD-TDP 'continuum' cases with GRN mutation and with coexisting MND or HS may have a more distinctive pathology. PMID- 20012112 TI - Significance of eukaryotic translation elongation factor 1A in tobacco mosaic virus infection. AB - Eukaryotic translation elongation factor 1A (eEF1A) has been shown to interact with both the viral RNA-dependent RNA polymerase and the 3'-terminal genomic RNA of tobacco mosaic virus (TMV). In this study, we demonstrated that the down regulation of eEF1A mRNA levels by virus-induced gene silencing using potato virus X vector dramatically reduced the accumulation of TMV RNA and the spread of TMV infection. The translation activity of the eEF1A-silenced Nicotiana benthamiana leaves was not severely affected. Collectively, these results suggest an essential role of eEF1A in TMV infection. PMID- 20012111 TI - Study of the neural basis of striatal modulation of the jaw-opening reflex. AB - Previous experimental data from this laboratory demonstrated the participation of the striatum and dopaminergic pathways in central nociceptive processing. The objective of this study was to examine the possible pathways and neural structures associated with the analgesic action of the striatum. The experiments were carried out in rats anesthetized with urethane. The jaw-opening reflex (JOR) was evoked by electrical stimulation of the tooth pulp of lower incisors and recorded in the anterior belly of the digastric muscles. Intrastriatal microinjection of apomorphine, a nonspecific dopamine agonist, reduced or abolished the JOR amplitude. Electrolytic or kainic acid lesions, unilateral to the apomorphine-injected striatum, of the globus pallidus, substantia nigra pars reticulata, subthalamic nucleus and bilateral lesion the rostroventromedial medulla (RVM), blocked the inhibition of the JOR by striatal stimulation. These findings suggest that the main output nuclei of the striatum and the RVM may be critical elements in the neural pathways mediating the inhibition of the reflex response, evoked in jaw muscles by noxious stimulation of dental pulp. PMID- 20012110 TI - Modulation of motorcortical excitability by methylphenidate in adult voluntary test persons performing a go/nogo task. AB - This study investigated the interaction between motorcortical excitability (short interval cortical inhibition, intracortical facilitation and long interval cortical inhibition), different requirement conditions [choice reaction test (CRT), attention/go/nogo], and their pharmacological modulation by methylphenidate (MPH) in normal healthy adults (n = 31) using a transcranial magnetic stimulation paradigm. MPH was administered in a dosage of 1 mg/kg body weight, maximum 60 mg. Additionally, serum level and clearance of MPH were controlled. The statistical analysis of variance revealed a significant three-way interaction of 2 (MPH) x 3 (CRT) x 6 (ISI) predicting motor evoked potential amplitudes (P = 0.032, MPH none and full dose, n = 31). In order to compare effects of dosage an additional between-subjects factor (half vs. full MPH dose) was introduced. None of the interactions involving this between-subject factor reached statistical significance. Exploring interactions with MPH only, a 3 (MPH none, half and full dose) x 3 (CRT) x 6 (ISI) analysis of variance revealed significant two-way interactions for MPH x ISI (P = 0.040) and condition x ISI (P < 0.001, n = 18). Effects observed for MPH were strongest on facilitatory processes, weaker for intracortical inhibition. In sum, MPH seems to interact via striato-thalamo-cortical pathways with original motorcortical processes (ISI), to a lesser extent with task-dependent or behavioral parameters (CRT). PMID- 20012114 TI - Potential anticancer application of polyamine oxidation products formed by amine oxidase: a new therapeutic approach. AB - The polyamines spermine, spermidine and putrescine are ubiquitous cell components. These molecules are substrates of a class of enzymes that includes monoamine oxidases, diamine oxidases, polyamine oxidases and copper-containing amine oxidases. Amine oxidases are important because they contribute to regulate levels of mono- and polyamines. In tumors, polyamines and amine oxidases are increased as compared to normal tissues. Cytotoxicity induced by bovine serum amine oxidase (BSAO) and spermine is attributed to H(2)O(2) and aldehydes produced by the reaction. This study demonstrated that multidrug-resistant (MDR) cancer cells (colon adenocarcinoma and melanoma) are significantly more sensitive than the corresponding wild-type (WT) ones to H(2)O(2) and aldehydes, the products of BSAO-catalyzed oxidation of spermine. Transmission electron microscopy (TEM) observations showed major ultrastructural alterations of the mitochondria. These were more pronounced in MDR than in WT cells. Increasing the incubation temperature from 37 to 42 degrees Celsius enhances cytotoxicity in cells exposed to spermine metabolites. The combination BSAO/spermine prevents tumor growth, particularly well if the enzyme has been conjugated to a biocompatible hydrogel polymers. Since both wild-type and MDR cancer cells after pre-treatment with MDL 72527, a lysosomotropic compound, are sensitized to subsequent exposure to BSAO/spermine, it is conceivable that combined treatment with a lysosomotropic compound and BSAO/spermine would be effective against tumor cells. It is of interest to search for such novel compounds, which might be promising for application in a therapeutic setting. PMID- 20012115 TI - Synthetic polyamines: an overview of their multiple biological activities. AB - The binding of polyamines to a variety of receptors and other defined recognition sites has been widely reported. It is well-known that polyamines interact with aspartate, glutamate, and aromatic residues of a given receptor and/or enzyme mainly through the formation of ion bonds, since at physiological pH, protonation of amino groups is nearly complete. From this, the hypothesis arises that a polyamine may be a universal template able to recognize different receptor systems. This hypothesis suggests that both affinity and selectivity may be fine tuned by inserting appropriate substituents onto the amine functions and by varying the methylene chain lengths between them on the polyamine backbone. In this paper, we detail several application of this design strategy aimed at discovering potent and selective polyamines able to bind neurotransmitter receptors and enzymes, such as muscarinic receptor subtypes, muscle-type nicotinic receptors and acethylcholinesterase. PMID- 20012116 TI - Metabolism of N-alkylated spermine analogues by polyamine and spermine oxidases. AB - N-alkylated polyamine analogues have potential as anticancer and antiparasitic drugs. However, their metabolism in the host has remained incompletely defined thus potentially limiting their utility. Here, we have studied the degradation of three different spermine analogues N,N'-bis-(3-ethylaminopropyl)butane-1,4 diamine (DESPM), N-(3-benzyl-aminopropyl)-N'-(3-ethylaminopropyl)butane-1,4 diamine (BnEtSPM) and N,N'-bis-(3-benzylaminopropyl)butane-1,4-diamine (DBSPM) and related mono-alkylated derivatives as substrates of recombinant human polyamine oxidase (APAO) and spermine oxidase (SMO). APAO and SMO metabolized DESPM to EtSPD [K(m(APAO)) = 10 microM, k(cat(APAO)) = 1.1 s(-1) and K(m(SMO)) = 28 microM, k(cat(SMO)) = 0.8 s(-1), respectively], metabolized BnEtSPM to EtSPD [K(m(APAO)) = 0.9 microM, k(cat(APAO)) = 1.1 s(-1) and K(m(SMO)) = 51 microM, k(cat(SMO)) = 0.4 s(-1), respectively], and metabolized DBSPM to BnSPD [K(m(APAO)) = 5.4 microM, k(cat(APAO)) = 2.0 s(-1) and K(m(SMO)) = 33 microM, k(cat(SMO)) = 0.3 s(-1), respectively]. Interestingly, mono-alkylated spermine derivatives were metabolized by APAO and SMO to SPD [EtSPM K(m(APAO)) = 16 microM, k(cat(APAO)) = 1.5 s(-1); K(m(SMO)) = 25 microM, k(cat(SMO)) = 8.2 s(-1); BnSPM K(m(APAO) )= 6.0 microM, k(cat(APAO)) = 2.8 s(-1); K(m(SMO)) = 19 muM, k(cat(SMO)) = 0.8 s(-1), respectively]. Surprisingly, EtSPD [K(m(APAO)) = 37 microM, k(cat(APAO)) = 0.1 s(-1); K(m(SMO)) = 48 microM, k(cat(SMO)) = 0.05 s( 1)] and BnSPD [K(m(APAO)) = 2.5 microM, k(cat(APAO)) = 3.5 s(-1); K(m(SMO)) = 60 microM, k(cat(SMO)) = 0.54 s(-1)] were metabolized to SPD by both the oxidases. Furthermore, we studied the degradation of DESPM, BnEtSPM or DBSPM in the DU145 prostate carcinoma cell line. The same major metabolites EtSPD and/or BnSPD were detected both in the culture medium and intracellularly after 48 h of culture. Moreover, EtSPM and BnSPM were detected from cell samples. Present data shows that inducible SMO parallel with APAO could play an important role in polyamine based drug action, i.e. degradation of parent drug and its metabolites, having significant impact on efficiency of these drugs, and hence for the development of novel N-alkylated polyamine analogues. PMID- 20012117 TI - Proteomic analysis of livers from a transgenic mouse line with activated polyamine catabolism. AB - We have generated a transgenic mouse line that over expresses the rate controlling enzyme of the polyamine catabolism, spermidine/spermine N (1) acetyltransferase, under the control of a heavy metal inducible promoter. This line is characterized by a notable increase in SSAT activity in liver, pancreas and kidneys and a moderate increase in the rest of the tissues. SSAT induction results in an enhanced polyamine catabolism manifested as a depletion of spermidine and spermine and an overaccumulation of putrescine in all tissues. To study how the activation of polyamine catabolism affects other metabolic pathways, protein expression pattern of the livers of transgenic animals was analyzed by two-dimensional polyacrylamide gel electrophoresis and mass spectrometry. A total of 23 proteins were shown to be differentially expressed in the transgenic from the wild-type animals. Many of the identified proteins showed expression patterns associated with polyamine catabolism activation. However, the expression pattern of other proteins, such as repression of GST pi and selenium binding protein 2 and 60 kDa heat-shock protein, could be explained by the overexpression of peroxisome proliferator-activated receptor gamma co-activator 1alpha in response to depleted ATP pools. The activation of the latter proteins is thought to lead to the improved insulin sensitivity seen in the MT-SSAT animals. PMID- 20012118 TI - Agmatine prevents the Ca(2+)-dependent induction of permeability transition in rat brain mitochondria. AB - The arginine metabolite agmatine is able to protect brain mitochondria against the drop in energy capacity by the Ca(2+)-dependent induction of permeability transition (MPT) in rat brain mitochondria. At normal levels, the amine maintains the respiratory control index and ADP/O ratio and prevents mitochondrial colloid osmotic swelling and any electrical potential (DeltaPsi) drop. MPT is due to oxidative stress induced by the interaction of Ca(2+) with the mitochondrial membrane, leading to the production of hydrogen peroxide and, subsequently, other reactive oxygen species (ROS) such as hydroxyl radicals. This production of ROS induces oxidation of sulfhydryl groups, in particular those of two critical cysteines, most probably located on adenine nucleotide translocase, and also oxidation of pyridine nucleotides, resulting in transition pore opening. The protective effect of agmatine is attributable to a scavenging effect on the most toxic ROS, i.e., the hydroxyl radical, thus preventing oxidative stress and consequent bioenergetic collapse. PMID- 20012120 TI - Features enhance the encoding of geometry. AB - Successful navigation within an environment requires that the traveler establish the correct heading--a process referred to as orienting. Many studies have now shown that humans and non-human animals can use the geometric properties of an enclosure to orient. In the present study, two groups of Clark's nutcrackers (Nucifraga columbiana) were trained, in a reference memory task, to find food hidden in one of four containers arranged to form a rectangular array. One group had unique objects placed next to each of the containers, whereas the second group had identical objects placed next to each of the containers. Here, I show for the first time that for the Clark's nutcracker, the distinctive properties of these objects enhanced the encoding of the array's geometry compared to the learning of geometric properties from an array of identical objects, which remained at chance after substantial amounts of training. Subsequent transformation tests showed that an object not associated with reward, but sharing the same geometric properties as the correct object, may have had inhibitory qualities. Furthermore, by systematically removing objects from the array, I show that although nutcrackers encoded the geometry of the array, they did not encode a complete featural representation of the objects within the array. PMID- 20012119 TI - Assay of T- and NK-cell subsets and the expression of NKG2A and NKG2D in patients with new-onset systemic lupus erythematosus. AB - This study aims to explore the percentage of T-cell and NK-cell subsets, the expression of NKG2A and NKG2D on CD3+ T cells and CD3-CD56+ NK cells on the total lymphocytes in new-onset systemic lupus erythematosus (SLE) patients, and explore clinical significance of these cell subsets. Thirty-two SLE patients and 32 normal controls were enrolled. Flow cytometry was used to count T- and NK-cell subsets and to detect the expression of NKG2A and NKG2D on CD3+ T cells and CD3 CD56+ NK cells in patients with new-onset SLE. Results show that CD4+ T (t = 2.04, P < 0.05), CD4+/CD8+ T cell (t = 2.66, P < 0.05), CD4+ CD25+ T (t = 2.48, P < 0.05), CD3+CD56+ natural killer T (NKT) (t = 40.05, P < 0.01), CD3-CD56+CD16+ NK-cell subsets (t = 3.50, P < 0.01) were significantly decreased, CD8+ T-cell subsets was significantly increased in patients with new-onset SLE (t = 3.80, P < 0.01), as compared with healthy controls. CD8+ T-cell subset was significantly increased in patients with vasculitis (t = 2.47, P < 0.05), and CD3-CD56+CD16+ NK was increased in patients with arthritis (t = 3.21, P < 0.01). However, no statistically significant correlation was found among different PBMC subsets and SLEDAI activity scores. Patients with SLE had a lower expression of NKG2A (U = 2.42, P < 0.05) as well as NKG2A/NKG2D ratio (t = 2.61, P < 0.05) and a higher expression of NKG2D (t = 2.21, P < 0.05) on CD3+ T cells, compared with normal controls. However, they had a higher expression of NKG2A (t = 2.59, P < 0.05) as well as NKG2A/NKG2D ratio (t = 49.45, P < 0.01) and a lower expression of NKG2D (t = 3.05, P < 0.01) on CD3-CD56+ NK cells. Taken together, the findings indicate the decreased CD4+ T-cell, CD4+/CD8+ T-cell, CD4+CD25+ T-cell, CD3+CD56+ NKT-, and CD3-CD56+CD16+ NK-cell subsets, increased CD8+ T-cell subsets as well as the abnormal expression of NKG2A and NKG2D on CD3+ T and CD3-CD56 + NK cells may play a role in the etiology of SLE. PMID- 20012121 TI - On the generality and limits of abstraction in rats and humans. AB - In this review, we address the question, central to cognition, of whether nonhuman animals such as rats are capable of extracting and extending information from a given learning situation to a new learning situation without generalizing through a physical dimension of the stimuli.This capacity underlies abstraction, which is a hallmark of human cognition and necessary for complex information processing such as language acquisition. We selectively review recent experiments with rats in which systematic changes in information processing of new stimuli are observed after training with different stimuli. These results strongly suggest that this capacity is present in rats. We also review two articles in which clear limitations to this capacity are detected. We conclude that, within specified limits, rats are capable of using prior experience when faced with a learning situation that involves new stimuli.We interpret this ability as a rudimentary form of abstraction. In the face of these provocative results, new theories of learning should be designed to account for these findings. PMID- 20012122 TI - Reported quality of randomized controlled trials in neglect rehabilitation. AB - The aim of this study is to assess the reported quality of randomized controlled trials (RCTs) on the effectiveness of neglect rehabilitation using a standardized scale. A search of seven electronic databases was carried out. Selected articles were scored using the PEDro scale and classified as high or low quality study both with the original cut off of 6 and a modified cut off of 5. A linear regression analysis between year of publication and quality rate was used to test whether the quality of the studies improved with time. A total of 18 RCTs were selected. Six articles (33.3%) and 10 articles (55.56%) were classified as having high quality when the original cut off or the modified cut off of the PEDro scale were used, respectively. Analysis shows no time-related changes in PEDro scores. The results show that reported quality is moderate for RCTs in neglect rehabilitation. PMID- 20012123 TI - Lack of cold pressor test-induced effect on visual-evoked potentials in migraine. AB - In patients with migraine, the various sensory stimulation modalities, including visual stimuli, invariably fail to elicit the normal response habituation. Whether this lack of habituation depends on abnormal activity in the sub-cortical structures responsible for processing incoming information as well as nociception and antinociception or on abnormal cortical excitability per se remains debateable. To find out whether inducing tonic pain in the hand by cold pressure test (CPT) alters the lack of visual-evoked potential (VEP) habituation in migraineurs without aura studied between attacks we recorded VEPs in 19 healthy subjects and in 12 migraine patients during four experimental conditions: baseline; no-pain (hand held in warm water, 25 degrees C); pain (hand held in cold water, 2-4 degrees C); and after-effects. We measured P100 amplitudes from six blocks of 100 sweeps, and assessed habituation from amplitude changes between the six sequential blocks. In healthy subjects, the CPT decreased block 1 VEP amplitude and abolished the normal VEP habituation (amplitude decrease to repeated stimulation) in patients with migraine studied between attacks; it left block 1 VEP amplitude and abnormal VEP habituation unchanged. These findings suggest that the interictal cortical dysfunction induced by migraine prevents the cortical changes induced by tonic painful stimulation both during pain and after pain ends. Because such cortical changes presumably reflect plasticity mechanisms in the stimulated cortex, our study suggests altered plasticity of sensory cortices in migraine. Whether this abnormality reflects abnormal functional activity in the subcortical structures subserving tonic pain activation remains conjectural. PMID- 20012124 TI - Population-based study of migraine in Spanish adults: relation to socio demographic factors, lifestyle and co-morbidity with other conditions. AB - The aim of this study was to estimate the prevalence of migraine in the general Spanish population and its association with socio-demographic and lifestyle factors, self-reported health status, and co-morbidity with other conditions. We analyzed data obtained from adults aged 16 years or older (n = 29,478) who participated in the 2006 Spanish National Health Survey (SNHS), an ongoing, home based personal interview which examines a nation-wide representative sample of civilian non-institutionalized population residing in main family dwellings (household) of Spain. We analyzed socio-demographic characteristics (gender, age, marital status, educational level, occupational status, and monetary monthly income); self-perceived health status; lifestyle habits (smoking habit, alcohol consumption, sleep habit, physical exercise, and obesity); and presence of other concomitant diseases. The 1-year prevalence of diagnosed migraine (n = 3,433) was 11.02% (95% CI 10.55-11.51). The prevalence was significantly higher among female (15.94%) than male (5.91%) and showed the highest value in the 31-50 years age group (12.11%). Migraine was more common in those of lower income (AOR 1.19, 95% CI 1.01-1.41) and who sleep <8 h/day (AOR 1.18, 95% CI 1.04-1.33). Furthermore, worse health status (AOR 2.04, 95% CI 1.76-2.36) and depression (AOR 1.82 95% CI 1.58-2.11) were related to migraine. Finally, subjects with migraine were significantly more likely to have comorbid conditions, particularly chronic (more than 6 month of duration) neck pain (AOR 2.31, 95% CI 1.98-2.68) and asthma (AOR 1.62, 95% 1.27-2.05). The current Spanish population-based survey has shown that migraine is more frequent in female, between 31 and 50 years and associated to a lower income, poor sleeping, worse health status, depression and several comorbid conditions, particularly chronic neck pain and asthma. PMID- 20012126 TI - Adolescents alcohol-use and economic conditions: a multilevel analysis of data from a period with big economic changes. AB - This paper examines how the unemployment rate is related to adolescent alcohol use and experience of binge drinking during a time period characterized by big societal changes. The paper uses repeated cross-sectional adolescent survey data from a Swedish region, collected in 1988, 1991, 1995, 1998, 2002 and 2005, and merges this with data on local unemployment rates for the same time periods. Individual level frequency of alcohol use as well as experience of binge drinking is connected to local level unemployment rate to estimate the relationship using multilevel modeling. The model includes municipality effects controlling for time invariant differences between municipalities as well as year fixed effects controlling for municipality-invariant changes over time in alcohol use. The results show that the unemployment rate is negatively associated with adolescents' alcohol use and the experience of binge drinking. When the unemployment rate increases, more adolescents do not drink at all. Regular drinking (twice per month or more) is, on the other hand, unrelated to the unemployment rate. Examining gender-differences in the relationship, it is shown that the results are driven by behavior in girls, whereas drinking among boys does not show any significant relationship with changes in the unemployment rate. PMID- 20012127 TI - Geographic distribution of physicians in Portugal. AB - The main goals of this paper are to (1) analyse the inequality in geographic distribution of physicians and its evolution, (2) estimate the determinants of physician density, and (3) assess the importance of competitive and agglomerative forces in location decisions. The analysis of the geographic distribution of physicians is based on the ratio of general practitioners (GPs) and specialists to 1,000 inhabitants. The inequality is measured using Gini indices, coefficients of variation, and physician-to-population ratios. The econometric models were estimated by ordinary least squares. The data used refer to 1996 and 2007. The impact of the growing number of physicians, and therefore potential increased competition, on geographic distribution during the period studied was small. Nonetheless, there is evidence of competitive forces acting on the dynamics of doctor localisation. Geographic disparities in physician density are still high, and appear to be due mainly to geographic income inequality. PMID- 20012125 TI - Does sumatriptan cross the blood-brain barrier in animals and man? AB - Sumatriptan, a relatively hydrophilic triptan, based on several animal studies has been regarded to be unable to cross the blood-brain barrier (BBB). In more recent animal studies there are strong indications that sumatriptan to some extent can cross the BBB. The CNS adverse events of sumatriptan in migraine patients and normal volunteers also indicate a more general effect of sumatriptan on CNS indicating that the drug can cross the BBB in man. It has been discussed whether a defect in the BBB during migraine attacks could be responsible for a possible central effect of sumatriptan in migraine. This review suggests that there is no need for a breakdown in the BBB to occur in order to explain a possible central CNS effect of sumatriptan. PMID- 20012128 TI - Colorectal cancer management in the United Kingdom: current practice and challenges. AB - This paper explores the current situation of colorectal cancer (CRC) management in the UK. Colorectal cancer is the third leading cause of cancer death in the UK, and the second and third leading cause of new cancer cases in women and men, respectively. These figures have increased over the past decade and, coupled with poorer survival rates compared to the European average and an ageing population, have recently led to the implementation of national screening in 2007-2009. Staffing adjustments are being made in preparation for national CRC-related demands, and national treatment guidelines are currently being revised. Access to new targeted treatments, however, is very limited due to negative NICE guidance and access is granted to only a select few. Targets have been created with regards to time to diagnosis and treatment, which are generally being met by the majority of hospitals. Overall, the focus on screening activities in the UK will hopefully result in fewer end-stage CRC cases and associated costlier treatments. PMID- 20012130 TI - The burden of colorectal cancer: prevention, treatment and quality of services. PMID- 20012129 TI - The dynamics of colorectal cancer management in 17 countries. AB - This paper discusses the current care management arrangements for colorectal cancer (CRC) in 16 OECD countries plus the Russian Federation by analysing data sources, the uptake of screening and surveillance, the available capacity in endoscopy services, the treatment pathways in medical treatment, as well as the type and availability of pharmaceutical care. The paper highlights significant variations in practice across the 17 countries. Common themes emerge from each of these practices and standards in terms of political interest in policies and awareness of CRC (both of which need to be enhanced), affordability (in terms of scarcity of resources in some countries and out-of-pocket payments for parts of the overall treatment process), access (in terms of the significant variation that has been observed within and across countries with regard to diagnostics, treatment and certain pharmaceuticals) and quality of CRC services (which may arise due to variations in treatment and pharmaceutical guidelines as well as minimal monitoring). When considering policy options for the future, it is important to, first, improve data collection both within as well as across countries through international co-operation; second, it is critical to have greater national and international support for cancer screening activities proven to be effective and cost-effective; third, endoscopy capacity in individual countries needs to be improved, also allowing more choice to ensure timely diagnosis, regardless of screening activities; fourth, public and political awareness needs to be enhanced as it is the key to improving CRC outcomes; fifth, where appropriate, to give consideration to the principles of equity, human dignity and disease severity, among others, when deciding on the uptake of new (targeted) treatments, rather than base decisions solely on cost-effectiveness criteria; and sixth, to firm up national guidelines including screening, diagnosis, treatment, pharmaceutical treatments and surveillance, with a view to enhancing their timeliness, evidence-base and free access to all. PMID- 20012131 TI - Colorectal cancer care in Denmark: status and dilemmas. AB - The area of cancer care has received significant political attention in Denmark over the past decades. Colorectal cancer (CRC) has a relatively high burden of disease and high mortality. This paper describes the main initiatives to improve CRC treatment in Denmark. It highlights the introduction of national "cancer pathways" based on clinical guidelines. The pathways specify ideal patient pathways through the system and include standards for waiting times and care processes. The comprehensiveness and high quality of Danish patient registers will aid in the implementation. Yet a number of outstanding challenges are also identified. The most important include a shortage of nursing and medical specialists for cancer care, and issues of co-ordination between primary care, hospital care and follow-up services. PMID- 20012132 TI - Colorectal cancer in Russia. AB - Using peer-reviewed publications and official government documents, this paper covers three important issues concerning colorectal cancer (CRC) in Russia: (1) CRC epidemiology, (2) screening for CRC, (3) medical care for patients with CRC. Colon and rectal cancer are registered separately in Russia. When colon and rectal cancers are considered together, CRC is the second cause of cancer death for women (after breast cancer) and the third for men (after lung and stomach cancer), while both incidence and mortality have increased over the past decade. About a quarter of all new colon and rectal cancer cases are diagnosed at Stage IV and one-third of patients with newly diagnosed disease die in the first year after diagnosing, however, poor data collection and collation presents problems with reliability of cancer statistics. Screening for CRC is not included in the national programme of preventive medical examination. There are no data about treatment effectiveness and access to innovative drugs in common practice. New drugs for CRC are included in the documents that regulate access to care, notably, drug lists and standards of medical care. However, many cancer patients are not included in any reimbursement programme and have to pay for the drugs, and there are problems with drug supply even for patients who are entitled to reimbursement. A national programme for oncology care development was launched in 2009 but it does not cover routine treatment. PMID- 20012133 TI - Cancer management and reimbursement aspects in Germany: an overview demonstrated by the case of colorectal cancer. AB - This paper provides an overview of cancer management, particularly with respect to colorectal cancer (CRC), in Germany. The information presented reflects findings from the peer-review literature and government documentation, as well as interviews with cancer and CRC specialists. Topics such as epidemiology, expenditure, CRC screening, pharmaceutical and non-pharmaceutical treatment are discussed in this paper. Furthermore, insights into the German reimbursement system with regard to cancer management as well as regulatory aspects are presented. PMID- 20012134 TI - Colocteral cancer management and prevention policies in Greece. AB - Despite lower cancer mortality rates in Greece compared to other European countries, colorectal cancer (CRC) is the third most important cause of cancer mortality. Given the significant economic and societal impact of CRC in Greece, this paper focuses on CRC data resources, disease management and the existing prevention policies. Numerous initiatives have taken place for the collection of data and the creation of cancer registries, however, they currently remain incomplete. Despite universal access of the Greek population to health services provision, structural problems of the national health system have imposed organisational barriers to the geographical distribution of health resources. National invitational CRC screening programmes focusing on early detection of the disease as well as guidelines for its management are missing. All novel cancer treatments are fully reimbursed. Post-treatment surveillance guidelines for high risk patients are very limited and depend solely on health providers' decisions. A National Cancer Plan (NCP) had been announced during the 2008-2012, but is still in the planning phases. Under the proposed NCP, the longevity of cancer data collection seems to be the critical step in monitoring and improving the performance of the health system. Detailed epidemiological data will give the possibility of constructing an effective prevention policy, will reduce socio economic inequalities in the access to CRC treatment and lighten the differences observed in health outcomes. The full implementation of the NCP constitutes the most significant investment in cancer management that has ever taken place in Greece. PMID- 20012135 TI - The disease burden of colorectal cancer in Hungary. AB - The aim of the paper is to give an overview of the epidemiology, treatment pattern and quality, as well as policy issues and disease burden of colorectal cancer (CRC) in Hungary. Colorectal cancer is the second most common cause of cancer-related death in both males and females in Hungary. The Hungarian Cancer Registry collects data on the epidemiological characteristics of CRC. Two pilot programmes (1997/1998 and 2003/2004) were conducted for population-based screening of CRC using both immunological and guaiac faecal occult blood testing (FOBT). The National Health Insurance Fund Administration (NHIFA) spends altogether 38.9 million a year on the treatment of CRC. It is hoped that the introduction of an accepted and cost-effective screening programme for CRC can reduce the high CRC burden in Hungary. PMID- 20012136 TI - Colorectal cancer in Italy: a review of current national and regional practice on screening and treatment. AB - Colorectal cancer (CRC) is one of the most prevalent and deadly cancers in Italy. Its burden is expected to remain significant in the coming years, although it is mostly a preventable disease. Prevention and screening programmes will play an important role in the fight against CRC. A national formal screening programme was introduced in the 2003-2005 and 2005-2007 national health programmes, leaving the planning and implementation, respectively, to each region and local health unit. In 2007, screening programmes covered 46.6% of the eligible population, with a higher coverage in the North (71.6%) and in the Centre (52.1%) than in the South (7%). The majority of programmes used the guaiac faecal occult blood test (FOBT) as first-line test. Only few programmes used the flexible sigmoidoscopy, or a combination of both tests. The quality and efficacy of the screening programmes are evaluated using ad hoc indicators with acceptable and desirable targets. In Italy, there are formal guidelines for population-based and opportunistic screening, diagnosis, surgery, adjuvant and neoadjuvant treatments and surveillance, differentiating colon cancer from rectal cancer and advanced CRC. Guidelines are updated yearly. Overall, Italy is well positioned in the fight against CRC. Although many regions lag behind in the uptake of screening programmes, they are in the process of introducing them. PMID- 20012137 TI - Colorectal cancer in Mexico: should a middle income country invest in screening or in treatment? AB - Mexico, like many other middle-income countries, is experiencing a demographic and epidemiological transition resulting in an older population suffering from chronic diseases. At the present time, cancer is the second cause of death in the country. Until recently, cervical carcinoma was the most frequent type of cancer in the country, however, the incidence of breast, prostate and colon cancers is growing. The demand for health care and health expenditure represented by cancer treatment challenges the limited resources the country has, particularly as patients seek treatment in advanced stages of the disease. Interestingly enough, these types of cancers could be detected in the early stages with rather simple screening procedures. The purpose of this paper is to describe the Mexican health system, and the impact of its fragmentation on access to medicines. Focusing on colorectal cancer (CRC), we describe its epidemiology, screening procedures and the inequities in health care access for these patients. PMID- 20012138 TI - The status of colorectal cancer care in the Netherlands: past, present and future. AB - Cancer is now the primary cause of death in the Netherlands, with colorectal cancer (CRC) being responsible for a significant portion of cancer incidence (14%) and mortality (11.8%) in men and women. An aging population and the possibility of national CRC screening, currently under discussion, will likely increase this burden. Despite switching in 2006 to obligatory subsidized private health insurance, health expenditure continues to rise, along with increased cancer and drug expenditures. Despite rising drug expenditures, new targeted biological treatments are available under centrally subsidized provisions. CRC treatment guidelines are available, published online, regularly updated, and monitored for quality purposes. There are concerns regarding waiting times for diagnosis and treatment, as well as projected staff shortages; however, various plans are in place to combat these issues. Overall, it appears that the Netherlands is aware of the oncology issues affecting CRC management, and is contemplating the optimal course of action for the future. PMID- 20012139 TI - Colorectal cancer management in Poland: current improvements and future challenges. AB - Colorectal cancer (CRC) is the second most commonly identified malignant neoplasm diagnosed in men (12% of total cancers) and women (11%) in Poland, while CRC mortality is second in men (10.1%) and third in women (11.2%). The main reasons for increasing incidence and mortality are an aging population and an increase in environmental and lifestyle factors which may lead to cancer. In Poland there is a lack of historical (regularly published and accessible) data on cancer morbidity and survival rates. The Oncology Centre published cancer data for the first time in February 2009 the 2006, which, also for the first time, embraced the entire country. Oncology data collection in Poland is based on a network of 16 Regional Cancer Registries reporting to the Polish National Cancer Registry in Warsaw. An additional source of oncology data is the National Health Fund and the Central Statistical Office. The National Cancer Programme (2005) provides funding at ca 780 million euro, which includes amongst others the Early CRC Detection Programme to promote a free screening colonoscopy. Oncology services in Poland are funded almost entirely by public resources--the national budget as sustained by tax revenues (Ministry of Health) and the National Health Fund as sustained by the obligatory public health insurance contribution. Oncology expenditure covered by the national budget (Ministry of Health) and the National Cancer Programme in 2006 amounted to 44.8 million euro and 105.2 million euro, respectively. All these preventive, curative and organizational efforts have significantly improved access to efficient therapies (including radiotherapy) and diagnostic procedures in recent years in Poland, although, clearly, a lot remains to be done. PMID- 20012140 TI - Colorectal cancer in Portugal. AB - Increasing diagnosis and deaths caused by colorectal cancer (CRC) warrant closer examination of affected patients and focus on management of CRC in Portugal. In order to assess the extent and quality of the information available in Portugal, we first analyse Portuguese cancer registries and then the management of CRC by discussing the diagnostic process and medical care provided, especially pharmaceuticals. Other cancer indications are mentioned in order to illustrate current approaches of cancer in Portugal. Current national data on cancer patients are scarce and there are divergencies in methods of data collection and treatment amongst regional cancer registries. However, the available data is sufficient enough to understand the dimension of CRC, with age-standardised incidence of 37 per 100,000 and mortality of 31 per 100,000 annually. An ongoing project is restructuring health services to improve efficiency and quality, however, some problems exist. The regional inequity of access to health care facilities and long waiting times for diagnostic examinations and surgery are major examples. Despite the non-availability of clinical guidelines, a pilot screening programme started at the beginning of 2009 in the Centre Region of the country. It is hoped that this overview will provide the basis for discussion on improvements in CRC management in Portugal and lead to better outcomes. PMID- 20012141 TI - The challenge of colorectal cancer prevention in Spain. AB - Colorectal cancer (CRC) is the leading cause of all new cancer cases and the second major cause for all cancer deaths in Spain. CRC survival has improved over the past few decades and is now higher than the 5-years European average, but this is associated with improved treatment than to early detection. A number of screening pilot programs have been completed in various regions, likely leading to national implementation in the future. Treatment guidelines have been developed and implemented to set standards and reduce practice variability. Newer treatments are available, including laparoscopic surgery, adjuvant chemotherapy and the use of targeted treatments. Specific programs are in place for rural patients, as well as programs to reduce waiting times. Overall, Spain is aware of the issues raised by CRC oncology management and has acted to improve patient outcomes, adding screening to its arsenal will further impact survival rates. PMID- 20012142 TI - Colorectal cancer in France. AB - Colorectal cancer (CRC) presents a substantial health burden in France, as the third cause of new cancer cases after breast and prostate cancers and the second cancer mortality cause representing nearly 12% of cancer deaths and almost 10% of annual life year loss due to cancer. Its prognosis has dramatically improved since the 1970s due to earlier diagnosis and an increase in the number of cases eligible for surgery. The first cancer plan implemented for the period 2003-2006 led to further improvements via, amongst others, implementing a national cancer institute (INCA), developing research, improving the quality of care and facilitating access to innovative treatments. The second cancer plan (2009-2013) aims to expand the scope of the existing cancer plan including improving primary and secondary prevention. Indeed, CRC national screening was recently implemented across France, however, the rate of the target population actually screened remains low while earlier detection of CRC at early stages should lead to an increase in France's CRC survival rates. PMID- 20012143 TI - Chemical lumbar sympathectomy in plantar hyperhidrosis. AB - Plantar hyperhidrosis can cause great changes to an individual's quality of life. We described a case successfully treated by the minimally invasive method of percutaneous injection of 7.0% phenolic solution into the lumbar sympathetic chains. PMID- 20012144 TI - Postural change alters autonomic responses to breath-holding. AB - OBJECTIVE: We used breath-holding during inspiration as a model to study the effect of pulmonary stretch on sympathetic nerve activity. METHODS: Twelve healthy subjects (7 females, 5 males; 19-27 years) were tested while they performed an inspiratory breath-hold, both supine and during a 60 degrees head-up tilt (HUT 60). Heart rate (HR), mean arterial blood pressure (MAP), respiration, muscle sympathetic nerve activity (MSNA), oxygen saturation (SaO(2)) and end tidal carbon dioxide (ETCO(2)) were recorded. Cardiac output (CO) and total peripheral resistance (TPR) were calculated. RESULTS: While breath-holding, ETCO(2) increased significantly from 41 +/- 2 to 60 +/- 2 Torr during supine (p < 0.05) and 38 +/- 2 Torr to 58 +/- 2 during HUT60 (p < 0.05); SaO(2) decreased from 98 +/- 1.5% to 95 +/- 1.4% supine, and from 97 +/- 1.5% to 94 +/- 1.7% during HUT60 (p = NS). MSNA showed three distinctive phases, a quiescent phase due to pulmonary stretch associated with decreased MAP, HR, CO, and TPR; a second phase of baroreflex-mediated elevated MSNA which was associated with recovery of MAP and HR only during HUT60; CO and peripheral resistance returned to baseline while supine and HUT60; a third phase of further increased MSNA activity related to hypercapnia and associated with increased TPR. INTERPRETATION: Breath-holding results in initial reductions of MSNA, MAP, and HR by the pulmonary stretch reflex followed by increased sympathetic activity related to the arterial baroreflex and chemoreflex. PMID- 20012145 TI - Cardiac ectopy in chronic autonomic failure. AB - OBJECTIVE: Chronic autonomic failure (CAF), as in Parkinson disease (PD), multiple system atrophy (MSA), and pure autonomic failure (PAF), typically entails baroreflex failure, neurogenic orthostatic hypotension (NOH), and supine hypertension. The combination might predispose to cardiac ectopy, which in turn might predispose to syncope and falls during manipulations decreasing venous return to the heart. This study assessed whether CAF is associated with an increased prevalence of cardiac ectopy. METHODS: Recordings lasting > or = 15 min of the electrocardiogram, beat-to-beat heart rate, and continuous blood pressure were reviewed from a total of 97 CAF patients (34 PD + NOH, 48 MSA, 15 PAF) and 82 control subjects (41 PD without NOH, 33 non-parkinsonian patients, 8 healthy volunteers). Cardiac ectopy was considered present if there were at least two premature beats or an arrhythmia. RESULTS: Atrial ectopy was found in 74% of patients with PD + NOH, 68% with MSA, and 63% with PAF, prevalences 2-3 times those in PD without NOH (28%, p < 0.0001) or other controls (24%, p < 0.0001). Atrial ectopy was related to subject age (p < 0.0001), supine systolic pressure (p < 0.0001), and the orthostatic fall in systolic pressure (p = 0.0007) and inversely with baroreflex-cardiovagal gain (p = 0.005) and the orthostatic increment in plasma norepinephrine (p = 0.0004). In two PD + NOH patients, atrial ectopy was associated with documented sustained hypotension after the Valsalva maneuver; and in an MSA patient, acute atrial flutter/fibrillation was associated with sudden loss of consciousness. INTERPRETATION: CAF patients have a relatively high frequency of atrial ectopy, which might interact with baroreflex failure to increase morbidity from orthostatic hypotension. PMID- 20012146 TI - Characterisations of odorant-binding proteins in the tsetse fly Glossina morsitans morsitans. AB - Odorant-binding proteins (OBPs) play an important role in insect olfaction by mediating interactions between odorants and odorant receptors. We report for the first time 20 OBP genes in the tsetse fly Glossina morsitans morsitans. qRT-PCR revealed that 8 of these genes were highly transcribed in the antennae. The transcription of these genes in the antennae was significantly lower in males than in females and there was a clear correlation between OBP gene transcription and feeding status. Starvation over 72 h post-blood meal (PBM) did not significantly affect the transcription. However, the transcription in the antennae of 10-week-old flies was much higher than in 3-day-old flies at 48 h PBM and decreased sharply after 72 h starvation, suggesting that the OBP gene expression is affected by the insect's nutritional status. Sequence comparisons with OBPs of other Dipterans identified several homologs to sex pheromone-binding proteins and OBPs of Drosophila melanogaster. PMID- 20012147 TI - Plasma YKL-40 is elevated in patients with recurrent atrial fibrillation after catheter ablation. AB - AIM: To study plasma YKL-40 in patients with atrial fibrillation (AF) treated with radiofrequency (RF) catheter ablation and to assess the predictive role of plasma YKL-40 and its changes after restoration of sinus rhythm (SR). METHODS: Forty-six patients (mean age 55 years, range 31-81) with paroxysmal/persistent AF were treated with RF catheter ablation; Holter monitoring for 14 days was performed before ablation and after 3 months. Recurrent symptomatic AF or atrial tachycardia >10 min was considered failure, and the patients were offered a second ablation session. YKL-40 was determined in plasma samples taken prior to ablation and at follow-up visits up to 12 months after ablation. RESULTS: After a maximum of two ablations, 19 patients (41%) had SR without recurrence of AF after 12 months. The patients with no recurrence of AF had significantly lower baseline plasma levels of YKL-40 prior to ablation compared to patients with recurrence of AF (31 vs. 62 microg/l, P = 0.029). Plasma YKL-40 was not an independent predictor of recurrence of AF after ablation. No significant changes in plasma YKL-40 levels were seen from baseline to follow-up at 12 months. CONCLUSION: In patients with paroxysmal or persistent AF treated with catheter ablation, high plasma YKL-40 before ablation is associated with recurrence of AF. PMID- 20012148 TI - Lead identification and optimization of diaminopyrimidines as histamine H4 receptor ligands. AB - BACKGROUND: The human histamine H(4) receptor (hH(4)R) is a promising new target in the therapy of inflammatory or immune system diseases. METHODS: For the development of new hH(4)R ligands, a broad virtual screening was performed and two hits were identified. Their annelated heterocyclic core was optimized with regard to affinity and potency. RESULTS: Pharmacological characterization of the resulting diaminopyrimidines revealed different agonist and antagonist properties within the same scaffold. PMID- 20012149 TI - Involvement of histamine and histamine H2 receptors in nicotinamide-induced differentiation of human amniotic epithelial cells into insulin-producing cells. AB - OBJECTIVE AND DESIGN: Human amniotic epithelial cells (HAEC) resemble stem cells in their ability to differentiate into all three germ layers: endoderm, mesoderm, and ectoderm. Histamine receptors are expressed on HAEC. We examined the influence of histamine, and H(1) and H(2) antagonists on the generation of pancreatic islet beta-like cells from HAEC. MATERIALS AND METHODS: HAEC were isolated after term pregnancies (N = 12) and cultured for 14 days with nicotinamide (10 mM) in normoxia. Altogether, 72 cultures were established. Histamine (100 microM) effects were investigated with mepyramine (10 microM) or cimetidine (10 microM). After 7 and 14 days, the mean concentration of C-peptide (MCCP) in the culture medium was measured immunoenzymatically as a marker of pancreatic differentiation. RESULTS: MCCP was approximately threefold higher on day 14, compared to day 7. Histamine significantly increased MCCP, and more evident differences were observed after 7 days of culture than after 14 days. The mean percent increase +/-SEM in MCCP amounted to 142.19 +/- 21.7 and 79.03 +/- 12.35 compared to the controls on day 7 and 14, respectively. H(2) blockade significantly reduced histamine-related increase in MCCP, both on day 7 and 14 by 88.7 +/- 14.3 and 39.2 +/- 12.4%, respectively. H(1) receptor antagonist did not affect MCPP. CONCLUSION: Nicotinamide-induced pancreatic differentiation of HAEC into beta-like cells may be augmented, probably at its earlier stage, by histamine acting via H(2) receptors. PMID- 20012150 TI - Histamine oxidation in mouse adipose tissue is controlled by the AOC3 gene encoded amine oxidase. AB - INTRODUCTION: Histaminergic status can modify adipose tissue (AT) development: histamine-free mice exhibit visceral obesity, and treatments with H3-antagonists reduce body weight gain. However, direct histamine effects on AT remain poorly documented: it has been observed that histamine stimulates lipolysis in rodent adipocytes when its oxidation by amine oxidases (AOs) is blocked by inhibitors such as semicarbazide. OBJECTIVE: The aim of this work was to study the influence of AOC3 gene invalidation, encoding for semicarbazide-sensitive AO (SSAO), on histamine oxidation and on histamine lipolytic activity in AT. MATERIALS AND METHODS: Expression of AOC- and MAO-encoding genes was determined by real-type PCR in wild-type (WT) and SSAO-deficient (AOC3-KO) mice. Lipolysis was assessed by glycerol release in isolated adipocytes and AO activity by substrate-induced hydrogen peroxide formation in kidney, ileum and AT. RESULTS: The expression levels of the genes encoding AOC1, AOC2 or MAOA and MAOB were not modified in the AT of AOC3-KO mice. In WT mice, histamine oxidation was lower than that of the reference SSAO-substrate benzylamine in AT, but not in ileum. The order of magnitude regarding benzylamine oxidation was AT > ileum >> kidney. In AOC3-KO mice, benzylamine oxidation was abolished in all tissues, while histamine oxidation was abolished in AT but not in ileum. Histamine was inactive on lipolysis in WT but stimulated lipolysis in fat cells from AOC3-KO mice, without reaching the maximal intensity of beta-adrenergic stimulation. CONCLUSION: Histamine was mainly oxidized by diamine oxidase (AOC1 product) in intestine, but by SSAO (AOC3 product) in AT. When protected from its oxidation by SSAO in AT, histamine moderately activated lipolysis in adipocytes in AOC3-KO mice. PMID- 20012151 TI - Effect of bacterial wax esters in sensitized (A) and non-sensitized (NA) human blood cell incubates. PMID- 20012152 TI - Eddy current effects on a clinical 7T-68 cm bore scanner. AB - INTRODUCTION: Eddy currents induced by switching of magnetic field gradients can lead to distortions in short echo-time spectroscopy or diffusion weighted imaging. In small bore magnets, such as human head-only systems, minimization of eddy current effects is more demanding because of the proximity of the gradient coil to conducting structures. METHODS: In the present study, the eddy current behavior achievable on a recently installed 7 tesla-68 cm bore head-only magnet was characterized. RESULTS: Residual effects after compensation were shown to be on the same order of magnitude as those measured on two whole body systems (3 and 4.7 T), while using two to three fold increased gradient slewrates. PMID- 20012153 TI - A model for production, perception, and acquisition of actions in face-to-face communication. AB - The concept of action as basic motor control unit for goal-directed movement behavior has been used primarily for private or non-communicative actions like walking, reaching, or grasping. In this paper, literature is reviewed indicating that this concept can also be used in all domains of face-to-face communication like speech, co-verbal facial expression, and co-verbal gesturing. Three domain specific types of actions, i.e. speech actions, facial actions, and hand-arm actions, are defined in this paper and a model is proposed that elucidates the underlying biological mechanisms of action production, action perception, and action acquisition in all domains of face-to-face communication. This model can be used as theoretical framework for empirical analysis or simulation with embodied conversational agents, and thus for advanced human-computer interaction technologies. PMID- 20012154 TI - Single doses of all-trans-N-retinylacetamide slow down the ERG amplitude recovery after bleaching in rats. AB - To assess the safety and to quantify the effects of a single application of all trans-N-retinylacetamide on the rat retina measured by electroretinography (ERG). Brown Norway rats were assigned to either a control group (n = 13) or to one of the three groups treated with a single intra-peritoneal dose of all-trans-N retinylacetamide: 20 (n = 8), 5 (n = 7), or 1 mg/kg (n = 8). Full-field ERGs were performed 7 days before (baseline) and 12 h after treatment. Intensity-response relationship of b-wave amplitudes were evaluated in dark-adapted conditions using white stimuli (0.000003-0.3 cd.s/m(2)). Fast dynamics of rod sensitivity was assessed by a paired-flash paradigm; recovery dynamics of b-wave amplitudes after bleaching was followed for 70 min. Light-adapted ERGs were recorded for cone evaluation. No effects were found on either dark-adapted sensitivity or on fast rod recovery. However, drug treatment at 5 and 20 mg/kg significantly delayed ERG amplitude recovery after bleaching: 60 min after bleaching the b-wave amplitude was 21 + or - 9% (P < 0.05) and 66 + or - 10% (P < 0.05), respectively, compared to baseline. Recovery rates returned to normal 8 weeks after treatment. There were no changes in light-adapted ERG in any group. Systemic administration of a single dose of the visual cycle modulator all-trans-N-retinylacetamide reversibly delayed recovery of dark-adapted ERG amplitudes after bleaching, leaving other functions unchanged. This finding could make the compound potentially useful in experimental conditions or in specific diseases where the visual cycle is involved, such as retinitis pigmentosa or age-related macular degeneration. PMID- 20012155 TI - Postnatal hyperoxia and the developing rat retina: beyond the obvious vasculopathy. AB - Although a great deal of emphasis has been placed on the vasculopathy that is associated with oxygen-induced retinopathy (OIR), our studies also revealed significant and irreversible structural (retinal histology) and functional (scotopic and photopic electroretinograms) impairments that were significantly more severe in pigmented Long-Evans rats compared to the more commonly used albino Sprague Dawley rats. In the following pages, we will highlight what we have learned about the retinal pathophysiological processes of OIR taking place in strains of both rats with the hope that this will trigger investigations into new therapeutic strategies to complement those geared at preventing the vasculopathy. PMID- 20012156 TI - Osteochondroma of the convexity: pathologic-neuroimaging correlates of a lesion that mimics high-grade meningioma. AB - Intracranial chondromas are uncommon benign lesions usually attached to dura and located over the convexity of the skull. Osteochondromas are even rarer and additionally contain a benign bony component. Both lesions are reportedly difficult to distinguish from meningiomas on pre-operative neuroimaging studies, although few detailed pathologic-neuroimaging correlation studies have appeared in the literature, particularly for intracranial osteochondromas. A 33-year-old woman with a 4-year history of headaches presented with recent onset of left sided muscle spasms and weakness. Two days prior to admission to our hospital, neuroimaging studies had shown a large right convexity mass with unusual multifocal bright signal intensities throughout an otherwise isointense mass. The bright signals were interpreted as showing multifocal hemorrhage and the mass was felt to be a convexity meningioma. However, subsequent catheter angiography characterized the lesion as being avascular. The mass was resected en bloc. Extensive histological sectioning revealed a benign osteochondroma predominantly composed of lobules of hypocellular cartilage. Microdissection of the different components revealed that the multifocal, spicule-like bright foci interpreted as hemorrhage on neuroimaging studies were instead foci of benign bone containing metaplastic bone marrow with trilineage hematopoietic cell populations and adipose tissue. Centrally, the hilum of the lesion contained avascular loose connective tissue. No recent or remote hemorrhage was identified anywhere in the lesion. Rare convexity osteochondromas may be mistaken for high-grade meningiomas on neuroimaging studies; their avascular nature, coupled with their complex signal pattern can serve as clues to the correct pre-operative diagnosis. PMID- 20012157 TI - Desmoplastic infantile astrocytoma with benign histological phenotype and multiple intracranial localizations at presentation. AB - Desmoplastic infantile astrocytoma (DIA) and desmoplastic infantile ganglioglioma (DIG) are rare intracranial tumors that mostly occur in the first 2 years of life and involve superficial cerebral cortex. Despite the large size of these lesions and some worrisome histological and radiological features, prognosis is generally favorable after gross total resection. We report an original observation of a desmoplastic infantile astrocytoma in a 5-year-old boy with multiple localizations on initial presentation, including the unusual subtentorial region. Magnetic resonance imaging showed a temporal tumor with prepontine and interpeduncular extension, and two other distinct localizations in cisterna magna and left cerebellar hemisphere. Leptomeningeal enhancements were present around the basal cistern. The surgical samples, corresponding exclusively to subtentorial lesions, were devoid of anaplastic features; the temporal lesion was untouched because of the interpeduncular extension. Adjuvant chemotherapy was applied, with shrinkage of lesions. DIA and DIG are more generally unifocal at initial presentation. When the tumor is large, multilobular involvement is common, but multiple location of DIG is, on the contrary, very rare. Previously, only five cases of DIG/DIA located in two or more separate locations have been published. We report the sixth, and first noninfantile, case of DIA/DIG with multifocal initial presentation. PMID- 20012158 TI - Intravitreal bevacizumab to facilitate vitrectomy in idiopathic ischaemic retinal vasculitis. AB - To describe the use of intravitreal bevacizumab in idiopathic retinal vasculitis prior to vitrectomy and delamination. A young healthy male presented with idiopathic retinal vasculitis. His condition was refractory to panretinal photocoagulation and systemic steroids. The progression of his condition necessitated vitrectomy and delamination. Intravitreal bevacizumab was used to facilitate the surgical procedure, with effective results. Intravitreal bevacizumab can be an adjunct to vitrectomy in idiopathic retinal vasculitis. However, caution must be taken due to its ability to promote fibrosis and to produce further tractional and/or rhegmatogenous retinal detachment. Further controlled studies are required to understand its benefits in managing complicated idiopathic retinal vasculitis. PMID- 20012159 TI - HNCO-based measurement of one-bond amide 15N-1H couplings with optimized precision. AB - A pair of 3D HNCO-based experiments have been developed with the aim of optimizing the precision of measurement of (1)J(NH) couplings. Both pulse sequences record (1)J(NH) coupling evolution during the entire constant time interval that (15)N magnetization is dephasing or rephasing with respect to the directly bonded (13)C' nucleus, with (15)N(13)C' multiple quantum coherence maintained during the (13)C' evolution period. The first experiment, designed for smaller proteins, produces an apparent doubling of the (1)J(NH) coupling without any accompanying increases in line width. The second experiment is a J-scaled TROSY-HNCO experiment in which the (1)J(NH) coupling is measured by frequency difference between resonances offset symmetrically about the position of the downfield component of the (15)N doublet (i.e. the TROSY resonance). This experiment delivers significant gains in precision of (1)J(NH) coupling measurement compared to existing J-scaled TROSY-HNCO experiments. With the proper choice of acquisition parameters and sufficient sensitivity to acquire a 3D TROSY HNCO experiment, it is shown that (1)J(NH) couplings can be measured with a precision which approaches or exceeds the precision of measurement with which the frequency of the TROSY resonance itself can be determined. PMID- 20012160 TI - Thiamine reduces tissue lead levels in rats: mechanism of interaction. AB - Lead (Pb) toxicity has been a serious concern in industrialized societies because of its association with functional deficits in nervous, haematopoietic and renal systems. Several studies have shown beneficial effects of thiamine on Pb toxicity. It is speculated that Pb chelation by thiamine may be a possible mechanism. However, the exact nature of these interactions remained elusive. In the present study we have characterized the interaction of Pb with thiamine using UV-Vis as well as fluorescence spectroscopic methods and studied the effect of thiamine treatment on blood and tissue Pb levels during simultaneous or post exposure to Pb in rat model. The spectroscopic studies revealed that Pb interacts with the pyrimidine ring of thiamine, leading to its solubilization at physiological pH. Further, thiamine reduced the Pb levels in blood, kidney and bone during both simultaneous and post-exposure Pb treatment. Interestingly, thiamine appears to prevent the accumulation of Pb in bone during simultaneous treatment. Together these results suggest that pyrimidine ring of thiamine mediates its interaction with Pb, leading to the prevention of its accumulation and/or increased clearance from tissues. PMID- 20012161 TI - Antitumor and antioxidant role of Chrysaora quinquecirrha (sea nettle) nematocyst venom peptide against Ehrlich ascites carcinoma in Swiss Albino mice. AB - This investigation aims to evaluate the antitumor and antioxidant potential of Chrysaora quinquecirrha (sea nettle) nematocyst venom on Ehrlich ascites carcinoma (EAC) tumor model. Tumor was induced in mice by intraperitoneal injection of EAC cells. The antitumor effect of sea nettle nematocyst venom (SNV) peptide was evaluated by assessing in vitro cytotoxicity, survival time, hematological, and antioxidant parameters. Intraperitoneal injection of SNV peptide increased the survival time of the EAC-bearing mice. The SNV peptide brought back the altered levels of the hematological and antioxidant parameters in a dose dependent manner in EAC-bearing mice. The results were comparable to that of the result obtained from the animals treated with the standard drug 5 fluorouracil (20 mg/kg bw). Thus, present study revealed that SNV peptide possessed significant antitumor and antioxidant activity. PMID- 20012163 TI - Evaluation of in situ curable biodegradable polyurethanes containing zwitterion components. AB - Porous polyurethane networks containing covalently attached zwitterionic compounds dihydroxypolycaprolactone phosphorylcholine and 1,2-dihydroxy-N,N dimethylamino-propane sulfonate have been prepared and characterised. Three polymers were prepared by reacting methyl 2,6-diisocyanato hexanoate functionalised D: -glucose as prepolymer A with either polycaprolactone triol alone or with addition of 10 mol% zwitterion as prepolymer B. All polymer compositions were mixed with 10 wt% hydrated gelatin beads. The cured polymers with the gelatin beads showed compression strengths that were still suitable for use in articular cartilage repair. The incorporation of zwitterions yielded more hydrophilic polymers that showed increased water absorption and increased porosity. After four months degradation in phosphate buffered saline, the polymers containing zwitterions had approximately 50% mass loss compared with 30% mass loss for that with polycaprolactone triol alone. All polymers were non-toxic in chondrocyte-based assays. Subcutaneous implantation of these polymers into rats confirmed that the polymers degraded slowly. Only a very mild inflammatory response was observed and the polymers were able to support new, well vascularised tissue formation. PMID- 20012162 TI - Novel functions of photoreceptor guanylate cyclases revealed by targeted deletion. AB - Targeted deletion of membrane guanylate cyclases (GCs) has yielded new information concerning their function. Here, we summarize briefly recent results of laboratory generated non-photoreceptor GC knockouts characterized by complex phenotypes affecting the vasculature, heart, brain, kidney, and other tissues. The main emphasis of the review, however, addresses the two GCs expressed in retinal photoreceptors, termed GC-E and GC-F. Naturally occurring GC-E (GUCY2D) null alleles in human and chicken are associated with an early onset blinding disorder, termed "Leber congenital amaurosis type 1" (LCA-1), characterized by extinguished scotopic and photopic ERGs, and retina degeneration. In mouse, a GC E null genotype produces a recessive cone dystrophy, while rods remain functional. Rod function is supported by the presence of GC-F (Gucy2f), a close relative of GC-E. Deletion of Gucy2f has very little effect on rod and cone physiology and survival. However, a GC-E/GC-F double knockout (GCdko) phenotypically resembles human LCA-1 with extinguished ERGs and rod/cone degeneration. In GCdko rods, PDE6 and GCAPs are absent in outer segments. In contrast, GC-E(-/-) cones lack proteins of the entire phototransduction cascade. These results suggest that GC-E may participate in transport of peripheral membrane proteins from the endoplasmic reticulum (ER) to the outer segments. PMID- 20012164 TI - Ability of polyurethane foams to support placenta-derived cell adhesion and osteogenic differentiation: preliminary results. AB - In bone tissue reconstruction, the use of engineered constructs created by mesenchymal stem cells (MSCs) that differentiate and proliferate into 3D porous scaffolds is an appealing alternative to clinical therapies. Human placenta represents a possible source of MSCs, as it is readily available without invasive procedures and because of the phenotypic plasticity of many of the cell types isolated from this tissue. The scaffold considered in this work is a slowly degradable polyurethane foam (EF PU foam), synthesized and characterized for morphology and in vitro interaction with chorion mesenchymal cells (CMCs). These cells were isolated from human term placenta and cultured onto the EF PU foam using two different culture media (EMEM and NH osteogenic differentiation medium). Synthesized EF PU foam showed homogeneous pore size and distribution, with 89% open porosity. In vitro tests showed CMCs scaffold colonization, as confirmed by Scanning Electron Microscopy (SEM) observations and hematoxylin eosin staining. Alizarin Red staining revealed the presence of a small amount of calcium deposition for the samples treated with the osteogenic differentiation medium. Therefore, the proposed EF PU foam appears to stimulate cell adhesion in vitro, sustaining CMCs growth and differentiation into the osteogenic lineage. PMID- 20012166 TI - Topographically induced self-deformation of the nuclei of cells: dependence on cell type and proposed mechanisms. AB - Osteosarcoma-derived cell lines (SaOs-2, MG63) have recently been shown to deform their nucleus considerably in response to surface topography. Such a deformation had not been described previously. Here we present results on additional cell lines, including cancerous (OHS4, U2OS), immortalized (F/STRO-1(+)A and FHSO6) and healthy cells (HOP). The cancerous cells were found to deform extensively, the immortalized cells showed small deformations, whereas the healthy cells showed deformation only at short incubation times. These results suggest a strong link between the malignant transformation of cells and the state of the cytoskeletal network. We propose mechanisms to explain the deformation in which the cytoskeleton either pushes down on the nucleus during spreading or pulls it down upon adhesion to the pillars. PMID- 20012165 TI - Amelogenin is phagocytized and induces changes in integrin configuration, gene expression and proliferation of cultured normal human dermal fibroblasts. AB - Fibroblasts are central in wound healing by expressing important mediators and producing and remodelling extracellular matrix (ECM) components. This study aimed at elucidating possible mechanisms of action of the ECM protein amelogenin on normal human dermal fibroblasts (NHDF). Amelogenin at 100 and 1000 microg/ml increased binding of NHDF via several integrins, including alphavbeta3, alphavbeta5 and alpha5beta1. Further, both surface interaction and cellular uptake of amelogenin by NHDF was observed using scanning and transmission electron microscopy. Gene microarray studies showed >8-fold up or down-regulation of genes, of which most are involved in cellular growth, migration and differentiation. The effect of amelogenin was exemplified by increased proliferation over 7 days. In conclusion, the beneficial effects of amelogenin on wound healing are possibly conducted by stimulating fibroblast signalling, proliferation and migration via integrin interactions. It is hypothesized that amelogenin stimulates wound healing by providing connective tissue cells with a temporary extracellular matrix. PMID- 20012167 TI - Hydrodynamic and species transfer simulations in the USP 4 dissolution apparatus: considerations for dissolution in a low velocity pulsing flow. AB - PURPOSE: To simulate the hydrodynamics in the flow-through (USP 4) dissolution apparatus and investigate the effects of hydrodynamics on mass transfer in a low velocity pulsing flow. METHODS: Computational fluid dynamics (CFD) was used to simulate the hydrodynamics and mass transfer in pulsing flow. Experimental flow visualisation was used to qualitatively confirm simulated hydrodynamic and mass transfer features. The experimental dissolution rate at 8 ml min(-1) (22.6 mm flow-through cell) was compared to the experimental dissolution rate in a free convection system. RESULTS: Simulations revealed periods of low velocity at all flow rates, evidence of boundary layer separation, and, at higher flow rates, residual fluid motion during zero inlet velocity periods. The simulated diffusion boundary layer thickness varied in certain regions over the course of the pulse. The experimental dissolution rate in the free convection system was faster than that at 8 ml min(-1) in the flow-through apparatus. CONCLUSIONS: A low velocity pulsing flow running counter to gravity inhibited the experimental dissolution rate compared to that in a free convection system. From the CFD simulations generated, simulation of both hydrodynamics and species transfer is recommended to characterise the influence of hydrodynamics on dissolution in a low velocity pulsing flow. PMID- 20012168 TI - Induced responses to herbivory and jasmonate in three milkweed species. AB - We studied constitutive and induced defensive traits (latex exudation, cardenolides, proteases, and C/N ratio) and resistance to monarch caterpillars (Danaus plexippus) in three closely related milkweed species (Asclepias angustifolia, A. barjoniifolia and A. fascicularis). All traits showed significant induction in at least one of the species. Jasmonate application only partially mimicked the effect of monarch feeding. We found some correspondence between latex and cardenolide content and reduced larval growth. Larvae fed cut leaves of A. angustifolia grew better than larvae fed intact plants. Addition of the cardenolide digitoxin to cut leaves reduced larval growth but ouabain (at the same concentration) had no effect. We, thus, confirm that latex and cardenolides are major defenses in milkweeds, effective against a specialist herbivore. Other traits such as proteases and C/N ratio additionally may be integrated in the defense scheme of those plants. Induction seems to play an important role in plants that have an intermediate level of defense, and we advocate incorporating induction as an additional axis of the plant defense syndrome hypothesis. PMID- 20012169 TI - Identification and molecular characterization of a Brachypodium distachyon GIGANTEA gene: functional conservation in monocot and dicot plants. AB - Developmental phase change and flowering transition are emerging as potential targets for biomass agriculture in recent years. The GIGANTEA (GI) gene is one of the central regulators that direct flowering promotion and phase transition. In this work, we isolated a GI gene orthologue from the small annual grass Brachypodium distachyon inbred line Bd21 (Brachypodium), which is perceived as a potential model monocot for studies on bioenergy grass species. A partial GI gene sequence was identified from a Brachypodium expressed sequence tag library, and a full-size gene (BdGI) was amplified from a Brachypodium cDNA library using specific primer sets designed through analysis of monocot GI gene sequences. The BdGI gene was up-regulated by light and cold. A circadian rhythm set by light dark transition also regulated the expression of the BdGI gene. The deduced amino acid sequence of the BdGI protein shares higher than 70% of sequence identity with the GI proteins in monocots and Arabidopsis. In addition, the BdGI protein is constitutively targeted to the nucleus and physically interacts with the ZEITLUPE (ZTL) and CONSTITUTIVE PHOTOMORPHOGENIC 1 (COP1) proteins, like the Arabidopsis GI protein. Interestingly, heterologous expression of the BdGI gene in a GI-deficient Arabidopsis mutant rescued efficiently the late flowering phenotype. Together, our data indicate that the role of the GI gene in flowering induction is conserved in Arabidopsis and Brachypodium. It is envisioned that the GI genes of bioenergy grasses as well as Brachypodium could be manipulated to improve biomass by engineering developmental timing of phase transitions. PMID- 20012170 TI - Salicylic acid differentially affects suspension cell cultures of Lotus japonicus and one of its non-symbiotic mutants. AB - Salicylic acid (SA) is known to play an important role in the interaction between plant and micro-organisms, both symbiotic and pathogen. In particular, high levels of SA block nodule formation and mycorrhizal colonization in plants. A mutant of Lotus japonicus, named Ljsym4-2, was characterized as unable to establish positive interactions with Rhizobium and fungi (NOD(-), MYC(-)); in particular, it does not recognize signal molecules released by symbiotic micro organisms so that eventually, epidermal cells undergo PCD at the contact area. We performed a detailed characterization of wild-type and Ljsym4-2 cultured cells by taking into account several parameters characterizing cell responses to SA, a molecule strongly involved in defense signaling pathways. In the presence of 0.5 mM SA, Ljsym4-2 suspension-cultured cells reduce their growth and eventually die, whereas in order to induce the same effects in wt suspension cells, SA concentration must be raised to 1.5 mM. An early and short production of nitric oxide (NO) and reactive oxygen species (ROS) was detected in wt-treated cells. In contrast, a continuous production of NO and a double-peak ROS response, similar to that reported after a pathogenic attack, was observed in the mutant Ljsym4-2 cells. At the molecular level, a constitutive higher level of a SA-inducible pathogenesis related gene was observed. The analysis in planta revealed a strong induction of the LjPR1 gene in the Ljsym4-2 mutant inoculated with Mesorhizobium loti. PMID- 20012171 TI - Biology of urothelial tumorigenesis: insights from genetically engineered mice. AB - Urothelium, one of the slowest cycling epithelia in the body, embodies a unique biological context for cellular transformation. Introduction of oncogenes into or removing tumor suppressor genes from the urothelial cells or a combination of both using the transgenic and/or knockout mouse approaches has provided useful insights into the molecular mechanisms of urothelial transformation and tumorigenesis. It is becoming increasingly clear that over-activation of the receptor tyrosine kinase (RTK) pathway, as exemplified by the constitutively activated Ha-ras oncogene, is both necessary and sufficient to initiate the low grade, non-invasive urothelial carcinomas. Dosage of the mutated Ha-ras, but not concurrent inactivation of pro-senescence molecules p16Ink4a and p19Arf, dictates whether and when the low-grade urothelial carcinomas arise. Inactivation of both p53 and pRb, a prevailing paradigm previously proposed for muscle-invasive urothelial tumorigenesis, is found to be necessary but insufficient to initiate this urothelial carcinoma variant. Instead, downregulation in p53/pRb co deficient urothelial cells of p107, a pRb family member, is associated with the genesis of the muscle-invasive bladder cancers. p53 deficiency also seems to be capable of cooperating with that of PTEN in eliciting invasive urothelial carcinomas. The genetically engineered mice have improved the molecular definition of the divergent pathways of urothelial tumorigenesis and progression, helped delineate the intricate crosstalk among different genetic alterations within a urothelium-specific context, identified new prognostic markers and novel therapeutic targets potentially applicable for clinical intervention, and provided in vivo platforms for testing preventive strategies of bladder cancer. PMID- 20012173 TI - Selection between Michaelis-Menten and target-mediated drug disposition pharmacokinetic models. AB - Target-mediated drug disposition (TMDD) models have been applied to describe the pharmacokinetics of drugs whose distribution and/or clearance are affected by its target due to high binding affinity and limited capacity. The Michaelis-Menten (M M) model has also been frequently used to describe the pharmacokinetics of such drugs. The purpose of this study is to investigate conditions for equivalence between M-M and TMDD pharmacokinetic models and provide guidelines for selection between these two approaches. Theoretical derivations were used to determine conditions under which M-M and TMDD pharmacokinetic models are equivalent. Computer simulations and model fitting were conducted to demonstrate these conditions. Typical M-M and TMDD profiles were simulated based on literature data for an anti-CD4 monoclonal antibody (TRX1) and phenytoin administered intravenously. Both models were fitted to data and goodness of fit criteria were evaluated for model selection. A case study of recombinant human erythropoietin was conducted to qualify results. A rapid binding TMDD model is equivalent to the M-M model if total target density R ( tot ) is constant, and R ( tot ) K ( D ) /(K ( D ) + C) ( 2 ) << 1 where K ( D ) represents the dissociation constant and C is the free drug concentration. Under these conditions, M-M parameters are defined as: V ( max ) = k ( int ) R ( tot ) V ( c ) and K ( m ) = K ( D ) where k ( int ) represents an internalization rate constant, and V ( c ) is the volume of the central compartment. R ( tot ) is constant if and only if k ( int ) = k ( deg,) where k ( deg ) is a degradation rate constant. If the TMDD model predictions are not sensitive to k ( int ) or k ( deg ) parameters, the condition of R ( tot ) K ( D ) /(K ( D ) + C) ( 2 ) << 1 alone can preserve the equivalence between rapid binding TMDD and M-M models. The model selection process for drugs that exhibit TMDD should involve a full mechanistic model as well as reduced models. The best model should adequately describe the data and have a minimal set of parameters estimated with acceptable precision. PMID- 20012172 TI - Urothelial carcinoma: stem cells on the edge. AB - Tumors are heterogeneous collections of cells with highly variable abilities to survive, grow, and metastasize. This variability likely stems from epigenetic and genetic influences, either stochastic or hardwired by cell type-specific lineage programs. That differentiation underlies tumor cell heterogeneity was elegantly demonstrated in hematopoietic tumors, in which rare primitive cells (cancer stem cells (CSCs)) resembling normal hematopoietic stem cells are ultimately responsible for tumor growth and viability. Because of the compelling clinical implications CSCs pose--across the entire spectrum of cancers--investigators applied the CSC model to cancers arising in tissues with crudely understood differentiation programs. Instead of relying on differentiation, these studies used empirically selected markers and statistical arguments to identify CSCs. The empirical approach has stimulated important questions about "stemness" in cancer cells as well as the validity and stoichiometry of CSC assays. The recent identification of urothelial differentiation programs in urothelial carcinomas (UroCas) supports the idea that solid epithelial cancers (carcinomas) develop and differentiate analogously to normal epithelia and provides new insights about the spatial localization and molecular makeup of carcinoma CSCs. Importantly, CSCs from invasive UroCas (UroCSCs) appear well situated to exchange important signals with adjacent stroma, to escape immune surveillance, and to survive cytotoxic therapy. These signals have potential roles in treatment resistance and many participate in druggable cellular pathways. In this review, we discuss the implications of these findings in understanding CSCs and in better understanding how UroCas form, progress, and should be treated. PMID- 20012174 TI - Index of consciousness and bispectral index values are interchangeable during normotension and hypotension but not during non pulsatile flow state during cardiac surgical procedures: a prospective study. AB - OBJECTIVES: Awareness under anesthesia is an avoidable complication during general anesthesia. Anesthetic depth monitors assist anesthesiologists in providing appropriate levels of anesthesia. Index of consciousness monitoring is a recently introduced monitor in the array of anesthesia depth monitors. The objective of this study was to assess the interchangeability of bispectral index, which is already in clinical use and the recently introduced index of consciousness techniques. The other objective was to assess this interchangeability during normotension, hypotension and during pulseless state in patients undergoing coronary artery bypass graft surgery. This study is a prospective observational study, conducted in a tertiary referral hospital. MEASUREMENTS AND MAIN RESULTS: Fifteen cardiac surgical patients undergoing off pump and conventional coronary artery bypass under cardiopulmonary bypass participated in the study. Bispectral index and index of consciousness monitoring were carried out simultaneously during various stages of consciousness, and assessed for interchangeability. Bland Altman plotting and 'mountain plot' were used to assess the interchangeability. Eleven in the cohort underwent off pump and the rest (n = 4) conventional coronary artery bypass surgery under cardiopulmonary bypass. A set of 887 data were obtained during the study period. The data were classified as those obtained during normotension, hypotension and pulseless state during cardiopulmonary bypass. RESULTS: 732 sets of data were obtained during normotension, 84 during hypotension and 71 during cardiopulmonary bypass. Overall interchangeability was good, suggested by low bias (0.96), high precision (0.54), r value of 0.7 and P value of <0.0001. It was found that the data obtained during normotension was also interchangeable, suggested by low bias (0.8), high precision (0.54) and r value of 0.7. The data obtained during hypotension was not as highly interchangeable as during normotension-bias 0.4, precision 1.66 and r value of 0.7. The analysis of value during cardiopulmonary bypass suggested non interchangeability (bias 3.87, precision 3.05, r value 0.3 and P value = 0.0067. CONCLUSIONS: The bispectral index and index of consciousness values may be interchangeable. The interchangeability is better appreciated during normotension and hypotension but not during non pulsatile state of cardiopulmonary bypass. PMID- 20012175 TI - Th17 cells facilitate the humoral immune response in patients with acute viral myocarditis. AB - BACKGROUND: Recently, the Th17 cell, a newly determined CD4+Th subset, was reported to participate in the inflammation of myocarditis combined with Th1 cells, and this study aimed to explore whether it was involved in the Th2 cell mediated humoral immunity in viral myocarditis. METHODS: A total of 34 patients, including 16 acute viral myocarditis (AVMC) and 18 dilated cardiomyopathy (DCM) having a history of AVMC, were enrolled for this study besides 18 healthy volunteers. RESULTS: The frequencies of Th17 and Th1 cells, especially Th17 cells in AVMC patients, while those of Th1 and Th2 cells, especially Th2 cells in DCM group, were all increased significantly compared with those in healthy volunteers (P < 0.01), with no changes of Th2 cells in AVMC and Th17 cells in DCM groups. The similar results were also observed in Th cell cytokines (IL-17, INF-gamma, and IL-4) and key transcript factors (RORgammat, T-bet, and GATA-3). Meanwhile, antiheart antibodies (AHA) of IgG type were found in 15 (93.8%) patients with AVMC and ten (55.6%) cases with DCM, accompanied by the higher expression of IL 17R on B cells and the frequencies of B cells than those in healthy controls (P < 0.01 in AVMC and P < 0.05 in DCM, respectively) who had no AHA. Furthermore, both of the B cell activities in AVMC and DCM groups were elevated and positively correlated to serum IL-17 (R = 0.66, P < 0.01) and IL-4 (R = 0.47, P < 0.05) respectively, with no correlation to INF-gamma. CONCLUSIONS: It was Th17 cells but not Th2 cells that helped the B cells to produce AHA in AVMC and not until at the late phase of viral myocarditis could Th2 cells play the important role in mediating humoral response. PMID- 20012176 TI - Therapeutic processes and perceived helpfulness of dang-ki (Chinese shamanism) from the symbolic healing perspective. AB - This study focuses on the therapeutic process and perceived helpfulness of dang ki, a form of Chinese shamanistic healing, in Singapore. It aims to understand the healing symbols employed in dang-ki, whether or not patients find them helpful and whether their perceived helpfulness can be explained by the symbolic healing model (Dow, Am Anthropol 88(1):56-69, 1986; Levi-Strauss, Structural anthropology. Basic Books, New York, 1963). Although many researchers have applied this model to explain the efficacy of shamanistic healings, they did not directly provide empirical support. Furthermore, the therapeutic process of a shared clinical reality as proposed by the model may be achievable in small-scale traditional societies that are culturally more homogeneous than in contemporary societies that are culturally more diversified due to globalization and immigration. Patients may hold multidimensional health belief systems, as biomedicine and alternative healing systems coexist. Thus, it would be interesting to see the relevance and applicability of the symbolic healing model to shamanistic healing in contemporary societies. In this study, ethnographic interviews were conducted with 21 patients over three stages: immediately before and after the healing and approximately 1 month later. The dang-ki healing symbols were identified by observing the healing sessions with video recording. Results show that dang-kis normally applied more than one method to treat a given problem. These methods included words, talismans and physical manipulations. Overall, 11 patients perceived their consultations as helpful, 4 perceived their consultations as helpful but were unable to follow all recommendations, 5 were not sure of the outcome because they had yet to see any concrete results and only 1 patient considered his consultation unhelpful. Although the symbolic healing model provides a useful framework to understand perceived helpfulness, processes such as enactment of a common meaning system and symbolic transformation are complex and dynamic, and may be carried over several healing sessions. PMID- 20012177 TI - Mitochondrial quality control: insights on how Parkinson's disease related genes PINK1, parkin, and Omi/HtrA2 interact to maintain mitochondrial homeostasis. AB - Alterations in mitochondrial homeostasis have been implicated in the etiology of Parkinson disease (PD) as demonstrated by human tissue studies, cell culture and in vivo genetic and toxin models. Mutations in the genes encoding PTEN-induced kinase 1 (PINK1), Omi/HtrA2 and parkin contribute to rare forms of parkinsonian neurodegeneration. Recently, each of these proteins has been shown to play a normal role in regulating mitochondrial structure, function, fission-fusion dynamics, or turnover (autophagy and biogenesis), promoting neuronal survival. Here, we review the biochemical mechanisms of mitochondrial protection conferred by each of these PD associated gene products in neurons, neuronal cell lines and other cell types. Potential molecular interactions and mitoprotective signaling pathways involving these three PD associated gene products are discussed in the context of mitochondrial quality control, in response to increasing levels of mitochondrial damage. We propose that PINK1, Omi/HtrA2 and parkin participate at different levels in mitochondrial quality control, converging through some overlapping and some distinct steps to maintain a common phenotype of healthy mitochondrial networks. PMID- 20012178 TI - Spectrum of Lyapunov exponents of non-smooth dynamical systems of integrate-and fire type. AB - We discuss how to characterize long-time dynamics of non-smooth dynamical systems, such as integrate-and-fire (I&F) like neuronal network, using Lyapunov exponents and present a stable numerical method for the accurate evaluation of the spectrum of Lyapunov exponents for this large class of dynamics. These dynamics contain (i) jump conditions as in the firing-reset dynamics and (ii) degeneracy such as in the refractory period in which voltage-like variables of the network collapse to a single constant value. Using the networks of linear I&F neurons, exponential I&F neurons, and I&F neurons with adaptive threshold, we illustrate our method and discuss the rich dynamics of these networks. PMID- 20012179 TI - Using self-determination theory to promote physical activity and weight control: a randomized controlled trial in women. AB - Behavior change interventions are effective to the extent that they affect appropriately-measured outcomes, especially in experimental controlled trials. The primary goal of this study was to analyze the impact of a 1-year weight management intervention based on self-determination theory (SDT) on theory-based psychosocial mediators, physical activity/exercise, and body weight and composition. Participants were 239 women (37.6 +/- 7.1 years; 31.5 +/- 4.1 kg/m(2)) who received either an intervention focused on promoting autonomous forms of exercise regulation and intrinsic motivation, or a general health education program (controls). At 12 months, the intervention group showed increased weight loss (-7.29%,) and higher levels of physical activity/exercise (+138 +/- 26 min/day of moderate plus vigorous exercise; +2,049 +/- 571 steps/day), compared to controls (P < 0.001). Main intervention targets such as more autonomous self-regulation (for treatment and for exercise) and a more autonomous perceived treatment climate revealed large effect sizes (between 0.80 and .96), favoring intervention (P < 0.001). Results suggest that interventions grounded in SDT can be successfully implemented in the context of weight management, enhancing the internalization of more autonomous forms of behavioral regulation, and facilitating exercise adherence, while producing clinically significant weight reduction, when compared to a control condition. Findings are fully consistent with previous studies conducted within this theoretical framework in other areas of health behavior change. PMID- 20012180 TI - One-carbon metabolism and CpG island methylator phenotype status in incident colorectal cancer: a nested case-referent study. AB - OBJECTIVE: We related prediagnostic plasma folate, vitamin B12, and total homocysteine concentrations, and the MTHFR 677C>T and 1298A>C polymorphisms, to the risk of colorectal cancer with and without the CpG island methylator phenotype (CIMP). METHODS: This was a nested case-referent study of 190 cases and double, matched referents from the large, population-based Northern Sweden Health and Disease Study. Using archival tumor tissue, promoter methylation in an eight gene panel was analyzed by MethyLight. RESULTS: A reduced risk of CIMP-low/CIMP high CRC (> or =1 gene methylated) was observed in subjects with very low plasma folate concentrations [multivariate odds ratio 2.96 (95% CI 1.24-7.08) for quintiles two to five versus one (lowest)]. With the exception of a reduced risk in MTHFR 677 TT-homozygotes, none of the other one-carbon variables were associated with the risk of CIMP-low/CIMP-high CRC. For CIMP-negative CRC, only the MTHFR polymorphisms were statistically significantly related to risk, inversely for 677C>T and positively for 1298A>C, but a tendency toward a reduced risk was observed in subjects with an adequate methyl availability, combining the plasma variables [multivariate odds ratio 0.61 (95% CI 0.32-1.15)]. CONCLUSION: Though limited by low power, these findings suggest the possibility of different roles for one-carbon metabolism in different pathways of colorectal tumorigenesis. PMID- 20012181 TI - Health and lifestyle behaviors among persons at risk of Lynch syndrome. AB - OBJECTIVE: The aim of this study was to evaluate health behaviors among patients with colorectal cancer (CRC) and their at-risk relatives prior to undergoing genetic counseling and testing for Lynch syndrome and to examine associations between health risk behaviors and specific demographic and psychological variables. METHODS: Participants included patients with CRC (n = 319) and their cancer-unaffected relatives (n = 110) who were enrolled in studies regarding Lynch syndrome genetic testing. Prior to undergoing genetic counseling or testing, participants completed a questionnaire including measures of demographic characteristics, health behaviors, cancer screening practices (Pap test, clinical breast exam, and mammogram), and psychological distress. RESULTS: Unaffected participants scored higher on a risk behavior index (RBI) than patients with CRC (1.7 (SD = 1.0) vs. 1.4 (SD = .09); p < .01). All female participants underwent cancer screening at rates similar to national data. Higher RBI scores were associated with being male, having less education, and age less than 50-years. CONCLUSIONS: We identified several health behaviors for potential intervention, including smoking, alcohol use, and diet. Genetic counseling offers a promising avenue for education and risk behavior reduction in persons at increased risk for cancer due to a familial or genetic predisposition, and a teachable moment to introduce lifestyle modifications. PMID- 20012183 TI - Proanthocyanidins and the risk of colorectal cancer in Italy. AB - Proanthocyanidins are a group of polymers of flavanols. Animal and in vitro studies suggest they decrease cancer risk, particularly of colorectal cancer. We used data from an Italian case-control study to investigate whether proanthocyanidins are related to colorectal cancer risk. Cases were 1,953 patients with incident, histologically confirmed colorectal cancer (1,225 colon cancers, 728 rectal cancers). Controls were 4,154 patients admitted for acute, non-neoplastic conditions. A reproducible and valid food frequency questionnaire was used. We estimated odds ratios (ORs) through multiple logistic regression models, including terms for potential confounding factors, and energy intake. A trend of decreasing risk with increasing intake of proanthocyanidins emerged for all classes except monomers. The OR for the highest vs. the lowest quintile of intake was 0.82 for monomers and dimers combined, 0.88 for monomers, 0.75 for dimers, 0.74 for all polymers with three or more mers, 0.84 for trimers, 0.80 for 4-6 mers, 0.79 for 7-10 mers, 0.69 for more than 10 mers, and 0.74 for total proanthocyanidins. The associations were apparently stronger for rectal than for colon cancer, in the absence of significant heterogeneity. These results may explain the protective effect of vegetables and fruit on colorectal cancer. PMID- 20012182 TI - Determinants of serum 25 hydroxyvitamin D levels in a nationwide cohort of blacks and non-Hispanic whites. AB - OBJECTIVE: To develop algorithms predicting serum 25 hydroxyvitamin D [s25(OH)D] for a large epidemiological study whose subjects come from large geographic areas, are racially diverse and have a wide range in age, skin types, and month of blood sample collection. This will allow a regression calibration approach to determine s25(OH)D levels replacing the more costly method of collection and analysis of blood samples. STUDY DESIGN AND SETTING: Questionnaire data from a subsample of 236 non-Hispanic whites (whites) and 209 blacks from the widely dispersed Adventist Health Study-2 (n = 96,000) were used to develop prediction algorithms for races separately and combined. A single blood sample was collected from each subject, at different times throughout the year. RESULTS: Models with independent variables age, sex, BMI, skin type, UV season, erythemal zone, total dietary vitamin D intake, and sun exposure factor explained 22 and 31% of the variance of s25(OH)D levels in white and black populations, respectively (42% when combined). UV season and erythemal zone determined from measured UV radiation produced models with higher R (2) than season and latitude. CONCLUSION: Combining races with a term for race and using variables with measured UV radiation capture the variance in s25(OH)D levels better than analyzing races separately. PMID- 20012184 TI - Depression and social anxiety in children: differential links with coping strategies. AB - Strategies that children use for coping with stressors are known to be related to emotional adjustment, but not enough is understood about specific links with social anxiety and depression. The present investigation tested differentiated associations of social anxiety and depression with specific types of coping strategies, and evaluated the direction of these associations over time. In Study 1, 404 children aged 8-13 years completed a coping scale modified from Kochendefer-Ladd and Skinner (Developmental Psychology 38:267-278, 2002) in order to evaluate factor structure and subscale internal consistency. In Study 2, 270 8 11-year-old children completed depression and social anxiety scales, a sociometric survey, and the coping scale from Study 1, with a follow-up timepoint 9 months later. In Study 1, factor analysis revealed six internally consistent coping subscales. In Study 2, social anxiety and depression were found to have distinctive longitudinal associations with subsequent coping strategies. Decreased problem-solving, social support-seeking, and distraction were uniquely predicted by depression but not by social anxiety. Internalising coping was a stronger outcome of social anxiety, and increased externalising was uniquely predicted by depression. There was also some evidence for a moderating role of peer relations. However, none of the coping strategies predicted changes in depression or social anxiety over the two timepoints. These results highlight the impact that emotional adjustment may have on children's coping strategies, and clarify important distinctions between social anxiety and depression in relation to coping. PMID- 20012185 TI - Molecular analysis of the replication program in unicellular model organisms. AB - Eukaryotes have long been reported to show temporal programs of replication, different portions of the genome being replicated at different times in S phase, with the added possibility of developmentally regulated changes in this pattern depending on species and cell type. Unicellular model organisms, primarily the budding yeast Saccharomyces cerevisiae, have been central to our current understanding of the mechanisms underlying the regulation of replication origins and the temporal program of replication in particular. But what exactly is a temporal program of replication, and how might it arise? In this article, we explore this question, drawing again on the wealth of experimental information in unicellular model organisms. PMID- 20012186 TI - Modulation of monoamine neurotransmitters in fighting fish Betta splendens exposed to waterborne phytoestrogens. AB - Endogenous estrogens are known to affect the activity of monoamine neurotransmitters in vertebrate animals, but the effects of exogenous estrogens on neurotransmitters are relatively poorly understood. We exposed sexually mature male fighting fish Betta splendens to environmentally relevant and pharmacological doses of three phytoestrogens that are potential endocrine disruptors in wild fish populations: genistein, equol, and beta-sitosterol. We also exposed fish to two doses of the endogenous estrogen 17beta-estradiol, which we selected as a positive control because phytoestrogens are putative estrogen mimics. Our results were variable, but the effects were generally modest. Genistein increased dopamine levels in the forebrains of B. splendens at both environmentally relevant and pharmacological doses. The environmentally relevant dose of equol increased dopamine levels in B. splendens forebrains, and the pharmacological dose decreased norepinephrine (forebrain), dopamine (hindbrain), and serotonin (forebrain) levels. The environmentally relevant dose of beta sitosterol decreased norepinephrine and dopamine in the forebrain and hindbrain, respectively. Our results suggest that sources of environmental phytoestrogens, such as runoff or effluent from agricultural fields, wood pulp mills, and sewage treatment plants, have the potential to modulate neurotransmitter activity in free-living fishes in a way that could interfere with normal behavioral processes. PMID- 20012187 TI - Assessment of pollution in sewage ponds using biomarker responses in wild African sharptooth catfish (Clarias gariepinus) in Tanzania. AB - The interactive effects of mixed pollutants in sewage wastewater on biomarker responses were investigated using wild male African sharptooth catfish (Clarias gariepinus) in Morogoro, Tanzania. A total of 58 fish were used, of which 21 were from Mindu dam (reference site) and 22, 9 and 10 from Mafisa, Mazimbu and Mzumbe sewage ponds, respectively. Liver somatic index (LSI) and gonadosomatic index (GSI) were significantly greater (two- to threefold) and (five- to sixfold), respectively, in fish from all sewage ponds. Haemoglobin concentration and gill filament 7-ethoxyresurufin O-deethylase (EROD) activities were significantly higher (1.2-fold and twofold, respectively) in fish from Mzumbe sewage ponds than in fish from Mindu dam, whereas liver EROD activity was significantly higher in fish from Mzumbe and Mafisa sewage ponds (5-fold). A HPLC method for determination of enzymatically formed p-nitrophenyl-glucuronide (PNPG) was developed and applied to measure UDP-glucuronosyl transferase (UGT) activities that was significantly higher in fish from all sewage ponds (2-2.5-fold) than in fish from Mindu dam. Kinetic characteristics and assay dependence of UGT were studied with microsomal preparations. Metallothionein (MT) content was significantly lower (three- to fourfold) in fish from sewage ponds than in fish from Mindu dam, and corresponded with cumulative levels of cadmium, lead and mercury. Condition factor, vitellogenin (Vtg), acetylcholinesterase (AChE) activities in plasma, eyes and brain, haematocrit, plasma protein and cytosolic glutathione S-transferase (GST) activities were comparable in fish from sewage ponds and Mindu dam. Although specific pollutants other than the metals were not identified by chemical analysis, application of a suite of biomarkers in C. gariepinus demonstrated that all sewage ponds were contaminated by pollutants of public health concern. PMID- 20012189 TI - Comparative efficacy of bromocriptine, cabergoline and thyroxine in inducing oestrus in bitches. AB - Forty bitches in anoestrus for more than six months from the last heat, with a serum progesterone level less than 1 ng/ml were subjected to oestrus induction trials using anti-prolactin drugs and levothyroxine, once daily orally for 20 consecutive days. The mean serum progesterone level among them was found to be 0.57 +/- 0.03 ng/ml. Out of 10 animals treated in each group, five (50%) in Group I (bromocriptine @ 50 microg/kg body weight), nine (90%) in Group II (cabergoline @ 5 microg/kg body weight), eight (80%) in Group III (thyroxine @10 microg/kg body weight) and seven (70%) in Group IV (thyroxine @ 5 microg/kg body weight) responded by evincing proestrual bleeding. The mean (+/-SEM) time taken from initiation of treatment to onset of proestrual bleeding in Groups I, II, III and IV was 28 +/- 3.39, 13.44 +/- 3.12 (P < 0.05), 24.50 +/- 3.18 and 33 +/- 2.21 days respectively. The mean (+/-SEM) duration of proestrus and oestrus in the treatment groups was 9.80 +/- 0.86, 10.11 +/- 0.68, 11.25 +/- 0.88 and 10.71 +/- 0.68 days and 7.60 +/- 0.24, 8 +/- 0.29, 8.5 +/- 0.63 and 7.85 +/- 0.46 days respectively. The conception rate in relation to the number of animals responding to oestrus induction in the treatment groups was 80%, 78%, 63% and 57%, respectively. The mean (+/-SEM) gestation length calculated from the last breeding date and litter size in the treatment groups varied from 60.50 +/- 1.55 to 64.00 +/- 0.82 days and 5.14 +/- 0.34 to 6.40 +/- 0.40 respectively. PMID- 20012188 TI - In vitro bioassay to detect reaginic antibodies from the serum of horses affected with recurrent airway obstruction. AB - In horses, Recurrent Airway Obstruction (RAO) is an allergic disease that involves IgE mediated Type I Hypersensitivity responses. The development of this type of allergy involves a series of events that begins with reaginic antibodies, mainly IgE and some IgG subclasses. These reaginic antibodies bind with high affinity, via the Fc portion, to FcepsilonRI receptors on the membrane of mast cells and basophils. Once bound, environmental allergens cross-link the antibodies, which results in mast cell degranulation leading to the production of histamine and other chemical mediators that act together to induce airway inflammation. RAO-affected horses present with coughing, respiratory distress, airway obstruction and poor performance. The aspect of the RAO has been extensively studied, yet the precise sequence of events is still not well understood. Therefore, this study proposes a bioassay for reaginic antibody detection from horse serum of RAO-affected individuals, in order to determine the etiology of disease, which mediate immediate type reactions. The technique involves measuring in vitro calcium mobilization in RBL-2H3 cells following incubation with horse serum from affected or unaffected horses and one of the RAO antigens (Aspergillus fumigatus). The results presented here demonstrate that 30% of RAO-affected horses react positively in this in vitro bioassay, whereas unaffected horses do not. This bioassay may facilitate further research on RAO and other allergic diseases in horses. PMID- 20012191 TI - Changes in freezing tolerance in hybrid poplar caused by up- and down-regulation of PtFAD2 gene expression. AB - In plant species, the level of polyunsaturated fatty acids (PUFAs) is essential for cold acclimation. To test whether changes in PUFA levels can lead to the alteration of freezing tolerance in poplar trees, we up- and down-regulated a Populus tomentosa Delta-12 fatty acid desaturase gene (PtFAD2) in the hybrid poplar (P. alba x P. glandulosa) clone 84 K. Real-time PCR results demonstrated that compared to untransformed control lines, the transcriptional level of PtFAD2 increased by up to 90% in over-expressing poplar lines (line OE-1) and decreased in down-regulated RNAi lines by up to 64% (line DR-1). As a result, the content of linoleic (C18:2) and linolenic (C18:3) unsaturated fatty acids (FAs) in total FAs increased by 7.5 and 3.9%, respectively, in the OE-1 line and decreased by 14.4 and 5.4% in the DR-2 line when compared to non-transgenic lines. After freezing treatment at -4 degrees C for 3 h without pre-cold acclimation, the survival rates of the PtFAD2-over-expressing cuttings were significantly higher (60% for OE-1) than those of non-transgenic plants (36.7%) and down-regulated lines (10% for DR-2). These results clearly demonstrate that the expression level of PUFAs substantially affected the freezing tolerance of hybrid poplar cuttings and could thus be utilized as an effective strategy to improve poplar anti freezing traits through genetic engineering biotechnology. PMID- 20012192 TI - Seven novel single nucleotide polymorphisms identified within river buffalo (Bubalus bubalis) lactoferrin gene. AB - The present study aimed at identifying single-nucleotide polymorphic (SNP) sites in different coding and non-coding regions of lactoferrin gene in Indian riverine buffaloes. A total of 102 animals from six different river buffalo breeds were screened at six bubaline lactoferrin gene loci. Single-strand conformation polymorphism (SSCP) analysis revealed monomorphic patterns at three loci LtfE2, LtfE11, and LtfE14 while a total of eight distinct patterns were observed in the other three loci viz. LtfE5, LtfE10, and LtfE16 which correspond to respective exons and their flanking regions. Sequence analysis of different SSCP variants revealed the presence of two SNP sites within the coding (exon 16) region and five SNP sites in flanking non-coding regions (intron 4 and intron 9). Both SNPs within exon 16 were found to be synonymous. The SNPs and haplotypes identified in the present study could serve as potential markers for association with susceptibility/resistance to mastitis in buffaloes. PMID- 20012190 TI - Transgenic expression of prothymosin alpha on zebrafish epidermal cells promotes proliferation and attenuates UVB-induced apoptosis. AB - This study generated a transgenic zebrafish line Tg(k18:Ptmaa-RFP) with overexpression of Prothymosin alpha type a (Ptmaa) in the skin epidermis. Red fluorescence first appears very weakly in the early stage, become stronger and mainly restricted in the nuclei of the epithelial cells from 3 dpf-larvae to adult fish. However, no evident morphological abnormalities were observed. Thus, overexpression of Ptmaa alone is not sufficient to cause disorganized growths or even cancer in zebrafish skin. Molecular and histological evidences showed that Tg(k18:Ptmaa-RFP) embryos have more proliferating cells in the pelvic fins [WT: 3.92 +/- 7.15; Tg(k18:Ptmaa-RFP): 38.00 +/- 10.87] and thicker skin [WT: 10.98 +/ 1.41 mum; Tg(k18:Ptmaa-RFP): 14.02 +/- 1.32 mum], indicating that overexpression of Ptmaa can promote proliferation. On the other hand, fewer apoptotic signals were found when Tg(k18:Ptmaa-RFP) embryos were exposed to UVB. Together with quantitative RT-PCR data, we suggest that UVB-induced epidermal cell apoptosis of zebrafish larvae can be attenuated by overexpression of Ptmaa through the enhancement of transcriptions of bcl2 mRNAs. Taken together, we conclude that overexpression of Ptmaa in zebrafish epidermal cells promotes proliferation and attenuates UVB-induced apoptosis but does not cause skin cancer. PMID- 20012193 TI - Age- and sex-dependent regression models for predicting the live weight of West African Dwarf goat from body measurements. AB - The relationships between live weight and eight body measurements of West African Dwarf (WAD) goats were studied using 211 animals under farm condition. The animals were categorized based on age and sex. Data obtained on height at withers (HW), heart girth (HG), body length (BL), head length (HL), and length of hindquarter (LHQ) were fitted into simple linear, allometric, and multiple regression models to predict live weight from the body measurements according to age group and sex. Results showed that live weight, HG, BL, LHQ, HL, and HW increased with the age of the animals. In multiple-regression model, HG and HL best fit the model for goat kids; HG, HW, and HL for goat aged 13-24 months; while HG, LHQ, HW, and HL best fit the model for goats aged 25-36 months. Coefficients of determination (R(2)) values for linear and allometric models for predicting the live weight of WAD goat increased with age in all the body measurements, with HG being the most satisfactory single measurement in predicting the live weight of WAD goat. Sex had significant influence on the model with R(2) values consistently higher in females except the models for LHQ and HW. PMID- 20012194 TI - The effect of dietary garlic supplementation on body weight gain, feed intake, feed conversion efficiency, faecal score, faecal coliform count and feeding cost in crossbred dairy calves. AB - Thirty-six crossbred calves (Holstein cross) of 5 days of age were used to study the effect of garlic extract feeding on their performance up to the age of 2 months (pre-ruminant stage). They were randomly allotted into treatment and control groups (18 numbers in each group). Performance was evaluated by measuring average body weight (BW) gain, feed intake (dry matter (DM), total digestible nutrient (TDN) and crude protein (CP)), feed conversion efficiency (FCE; DM, TDN and CP), faecal score, faecal coliform count and feeding cost. Diets were the same for the both groups. In addition, treatment group received garlic extract supplementation at 250 mg/kg BW per day per calf. Body weight measured weekly, feed intake measured twice daily, proximate analysis of feeds and fodders analysed weekly, faecal scores monitored daily and faecal coliform count done weekly. There was significant increase in average body weight gain, feed intake and FCE and significant decrease in severity of scours as measured by faecal score and faecal coliform count in the treatment group compared to the control group (P < 0.01). Feed cost per kilogramme BW gain was significantly lower in the treatment group compared to control group (P < 0.01). The results suggest that garlic extract can be supplemented to the calves for better performance. PMID- 20012195 TI - Comparison of artificial insemination with natural mating on smallholder farms in Thailand, and the effects of boar stimulation and distance of semen delivery on sow reproductive performance. AB - Following the implementation of artificial insemination (AI) services for smallholder pig farms, we investigated the reproductive performance after AI and its influencing factors. A small-scale boar station with an AI lab was established with two active boars having good genetics and free from reproductive diseases. Individual sow cards were used for reproductive data recording. A total of 171 sows on 92 farms situated within a radius of 50 km from the AI center were included in this study. Sows bred by AI (n = 121) were inseminated twice per estrus by two trained inseminators. A further 50 sows were mated by natural services using local rental boars. The impact of boar stimulation and distance from the AI center to the farm were also determined. Non-return (P = 0.02) and farrowing rates (P = 0.03) were higher for AI than for naturally bred sows (84.0% and 76.0% vs. 74.0% and 70.0% for AI and naturally bred, respectively). For sows bred by AI, boar stimulation increased non-return rate (84.1% vs. 70.0%; P = 0.09), farrowing rate (83.7% vs. 69.2%; P = 0.01) and litter size (11.2 +/- 2.3 vs. 9.7 +/- 1.7; P < 0.01). There was no effect on performance due to distance of semen transport. These results clearly indicate that sow performance on smallholder farms will improve if AI is utilized and boar stimulation is employed. PMID- 20012196 TI - Desulfovibrio oceani subsp. oceani sp. nov., subsp. nov. and Desulfovibrio oceani subsp. galateae subsp. nov., novel sulfate-reducing bacteria isolated from the oxygen minimum zone off the coast of Peru. AB - Two deltaproteobacterial sulfate reducers, designated strain I.8.1(T) and I.9.1(T), were isolated from the oxygen minimum zone water column off the coast of Peru at 400 and 500 m water depth. The strains were Gram-negative, vibrio shaped and motile. Both strains were psychrotolerant, grew optimally at 20 degrees C at pH 7.0-8.0 and at 2.5-3.5% NaCl (w/v). The strains grew by utilizing hydrogen/acetate, C(3-4) fatty acids, amino acids and glycerol as electron acceptors for sulfate reduction. Fumarate, lactate and pyruvate supported fermentative growth. Sulfate, sulfite, thiosulfate and taurin supported growth as electron acceptors. Both strains were catalase-positive and highly oxygen tolerant, surviving 24 days of exposure to atmospheric concentrations. MK6 was the only respiratory quinone. The most prominent cellular fatty acid was iso-17:1 omega9c (18%) for strain I.8.1(T) and iso-17:0-omega9c (14%) for strain I.9.1(T). The G+C contents of their genomic DNA were 45-46 mol%. Phylogenetic analysis of 16S rRNA and dsrAB gene sequences showed that both strains belong to the genus Desulfovibrio. Desulfovibrio acrylicus DSM 10141(T) and Desulfovibrio marinisediminis JCM 14577(T) represented their closest validly described relatives with pairwise 16S rRNA gene sequence identities of 98-99%. The level of DNA-DNA hybridization between strains I.8.1(T) and I.9.1(T) was 30-38%. The two strains shared 10-26% DNA-DNA relatedness with D. acrylicus. Based on a polyphasic investigation it is proposed that strains I.8.1(T) and I.9.1(T) represent a novel species for which the name Desulfovibrio oceani sp. nov. is proposed with the two subspecies D. oceani subsp. oceani (type strain, I.8.1(T) = DSM 21390(T) = JCM 15970(T)) and D. oceani subsp. galateae (type strain, I.9.1(T) = DSM 21391(T) = JCM 15971(T)). PMID- 20012197 TI - A pilot evaluation of the In SHAPE individualized health promotion intervention for adults with mental illness. AB - This pilot study examined whether participation in the individualized, community integrated In SHAPE health promotion program would result in improved physical activity and dietary behaviors, health indicators, and psychological functioning or symptoms in 76 adults with schizophrenia, bipolar disorder, major depression, or other mental disorders. Over a 9-month period, participation was associated with increased exercise, vigorous activity, and leisurely walking (P < .01), and a trend toward improved readiness to reduce caloric intake (P = .053). Participants demonstrated a reduction in waist circumference (P < .05), but no change in BMI. Satisfaction with fitness (P < .001) and mental health functioning (P < .05) improved, and severity of negative symptoms decreased (P < .01). This study demonstrated the feasibility and potential effectiveness of the In SHAPE program, which consisted of exercise and dietary modification. Based on these promising results, randomized controlled trials of the In SHAPE program are necessary to establish its effectiveness compared with usual care and alternative approaches to enhancing fitness. PMID- 20012198 TI - Effects of proton pump inhibitors on gastric emptying: a systematic review. AB - The proton pump inhibitor (PPI) is widely used for the treatment of gastroesophageal reflux disease, peptic ulcer diseases, and functional dyspepsia. The pathogenesis of these acid-related and/or functional upper gastrointestinal disorders is potentially associated with abnormal gastric emptying. To date, variable effects of PPIs on gastric emptying have been reported. Therefore, it is relevant to gather and analyze published information on this topic. A systematic literature search has been performed, showing that the delaying effect of PPIs on gastric emptying of solid meals is consistent, whereas the effect of PPIs on the emptying of liquids is inconsistent. The underlying mechanisms whereby PPIs may affect gastric emptying have been discussed, most of which still remain hypothetic. Gastric emptying of solids involves a process of peptic hydrolysis. PPIs impair the hydrolytic digestion by inhibiting acid-dependent peptic activity, thereby delaying the solid emptying. Gastric emptying of liquids largely depends on volume and energy density of intragastric contents. PPIs variably modify the volume and the energy density by reducing gastric fluid secretion, thereby modifying the liquid emptying in an unpredictable manner. Hypergastrinemia has been considered to delay gastric emptying, but it seems of minor importance in the regulation of gastric emptying during PPI use. The delayed emptying of solids due to PPI therapy may have clinical implications in the management of gastroesophageal reflux disease, functional dyspepsia, as well as diabetes. PMID- 20012199 TI - The effects of Project ALERT one year past curriculum completion. AB - School-based drug prevention curricula constitute the nation's most prevalent strategy to prevent adolescent drug use. We evaluated the effects of one such curriculum, Project ALERT, on adolescent substance use. In particular, we sought to determine if a single effect on 30-day alcohol use, noted shortly following the completion of the 2-year program, could be detected 1 year later. We also looked for delayed effects on other outcomes of interest, namely lifetime alcohol use, and 30-day and lifetime use of cigarettes, marijuana, and inhalants. We employed a randomized controlled trial that used school as the unit of assignment. Thirty-four schools with grades 6-8 from 11 states completed the study. Seventy-one Project ALERT instructors taught 11 core lessons to sixth graders and 3 booster lessons to seventh graders. Students were assessed prior to the onset of the intervention, as sixth graders, after the completion of the 2 year curriculum, as seventh graders, and again 1 year later as eighth graders. This paper examines data from the pretest and final posttest. Using hierarchical nonlinear modeling, we found that our earlier effect on 30-day alcohol use did not persist. Further, we continued to find no effects for lifetime alcohol use and both the lifetime and 30-day use of cigarettes, marijuana, and inhalants. Our findings do not support the long-term effectiveness of Project ALERT, when delivered to sixth graders. PMID- 20012200 TI - An extended model of reasoned action to understand the influence of individual- and network-level factors on African Americans' participation in HIV vaccine research. AB - In the United States, the number and proportion of HIV/AIDS cases among black/African Americans continue to highlight the need for new biomedical prevention interventions, including an HIV vaccine, microbicide, or new antiretroviral (ARV) prevention strategies such as pre-exposure prophylaxis (PrEP) to complement existing condom usage, harm reduction methods, and behavioral change strategies to stem the HIV epidemic. Although black/African Americans are disproportionately impacted by HIV/AIDS, their participation in HIV clinical research continues to have unique challenges. We theorize that interaction among multilevel factors creates ideal alignment for minority participation in HIV clinical studies. Thus, we initially set out to test an extended model of reasoned action with 362 participants to understand the interplay of sociopsychological and network-level considerations influencing minority participation in HIV prevention research efforts. In this study, we linked the intrapersonal dimensions of attitudes, beliefs, and normative concerns to community-level components, appraisal of involvement with the clinical research organization, an entity which operates within a networked structure of community partner agencies, and identification with coalition advocacy aims. Various participatory outcomes were explored including involvement in future HIV vaccine community functions, participation in community promotion of HIV vaccine research, and community mobilization. Three-stage least squares estimates indicated similar findings across three models. Significant effects demonstrate the importance of positive attitudes toward HIV vaccine research, favorable health research beliefs, perceived social support for participation, HIV/AIDS issue engagement, and perceived relevance of the clinical research site's mission and values. Identification of these nuanced pathway effects provides implications for tailored community program development. PMID- 20012201 TI - Photosynthetic and respiratory changes in leaves of poplar elicited by rust infection. AB - Poplars are challenged by a wide range of pathogens during their lifespan, and have an innate immunity system that activates defence responses to restrict pathogen growth. Large-scale expression studies of poplar-rust interactions have shown concerted transcriptional changes during defence responses, as in other plant pathosystems. Detailed analysis of expression profiles of metabolic pathways in these studies indicates that photosynthesis and respiration are also important components of the poplar response to rust infection. This is consistent with our current understanding of plant pathogen interactions as defence responses impose substantive demands for resources and energy that are met by reorganization of primary metabolism. This review applies the results of poplar transcriptome analyses to current research describing how plants divert energy from plant primary metabolism for resistance mechanisms. PMID- 20012202 TI - Assessing the needs and guiding the future: findings from the health needs assessment in 13 Asian American communities of Maryland in the United States. AB - There is lack of in depth data on health needs of diverse Asian American communities. We conducted 19 focus groups in 13 Asian American communities in Maryland in 2007. We developed a moderator's guide to collect qualitative data on health needs from 174 participants, and used MAX QDA to analyze data and code emergent themes. Cardiovascular disease related conditions, diabetes, and mental health were the top three health concerns. Weight concerns, cancer, arthritis, smoking, osteoporosis, and hepatitis B followed next. Many participants were not receiving preventive health service such as cancer screening due to a lack of access to health care or lack of awareness of preventive care. Additionally, under-represented communities lacked adequate health resources and advocacy, potentially due to a relatively shorter history of immigration and a small population. The results render support for awareness education on importance of preventive care and mental health. PMID- 20012203 TI - Association of depression, psycho-social stress and acculturation with respiratory disease among Puerto Rican adults in Massachusetts. AB - To assess associations between acculturation, depression, and self-reported stress score with reported diagnosis of respiratory disease (RD) in Puerto Rican adults, participants (N = 1,168) were identified from areas of high Hispanic density in the Boston, MA metropolitan area. Eligible participants were interviewed in the home by bilingual interviewers in either Spanish or English. Scales included topics ranging from general background to depressive symptomatology. Respiratory disease was self-reported and checked against prescribed medication. More than one-third (37.8%) of subjects reported doctor diagnosed RD. A final binary logistical regression model (N = 850), which was adjusted for potential confounders (sex, age, education, poverty) showed that RD was significantly associated with psychological acculturation (OR = 1.97, P = 0.005), depressive symptomatology (OR = 1.52, P = 0.03) high perceived stress score (OR = 1.97, P = 0.009), and current smoking (OR = 1.61, P = 0.03). Significant inverse associations included a high level of language acculturation (OR = 0.65, P = 0.03), light (OR = 0.67, P = 0.01) and moderate to heavy physical activity versus sedentary physical activity (OR = 0.40, P = 0.03). We found self reported physician diagnosed RD was associated with high perceived stress and depression, as well as higher levels of psychological acculturation. Longitudinal research is needed to determine if there is a causal pathway for these associations. PMID- 20012204 TI - Self-reported racial discrimination and substance use among Asian Americans in Arizona. AB - We examined associations between different forms of self-reported racial discrimination and current substance use (i.e., smoking, alcohol use, and controlled substance use) among Asian Americans living in Arizona. The data are from 271 Asian American adults participating in the 2008 Asian Pacific Arizona Inititative (APAZI) Survey, which is part of a larger collaboration between community members, organizational leaders, and researchers. Asian Americans treated like they were not American because of their race were at increased risk of tobacco use, after controlling for covariates, including age, gender, education, family income, health insurance, nativity status, and language, and other types of racial discrimination. Also, individuals treated differently because of their race were at increased risk of alcohol use and controlled substance use, after controlling for covariates and other types of racial discrimination. The results indicate that Asian Americans experience a wide range of racial discrimination types and some forms of racial discrimination may have greater associations with tobacco, alcohol, and controlled substance use than others. PMID- 20012205 TI - Boundary stiffness regulates fibroblast behavior in collagen gels. AB - Recent studies have illustrated the profound dependence of cellular behavior on the stiffness of 2D culture substrates. The goal of this study was to develop a method to alter the stiffness cells experience in a standard 3D collagen gel model without affecting the physiochemical properties of the extracellular matrix. A device was developed utilizing compliant anchors (0.048-0.64 N m(-1)) to tune the boundary stiffness of suspended collagen gels in between the commonly utilized free and fixed conditions (zero and infinite stiffness boundary stiffness). We demonstrate the principle of operation with finite element analyses and a wide range of experimental studies. In all cases, boundary stiffness has a strong influence on cell behavior, most notably eliciting higher basal tension and activated force (in response to KCl) and more pronounced remodeling of the collagen matrix at higher boundary stiffness levels. Measured equibiaxial forces for gels seeded with 3 million human foreskin fibroblasts range from 0.05 to 1 mN increasing monotonically with boundary stiffness. Estimated force per cell ranges from 17 to 100 nN utilizing representative volume element analysis. This device provides a valuable tool to independently study the effect of the mechanical environment of the cell in a 3D collagen matrix. PMID- 20012207 TI - Thrombosis of a bileaflet prosthetic mitral valve: a real-time three-dimensional transesophageal echocardiography perspective. AB - Rheumatic mitral valve disease being common in India, valve replacement and hence thrombosis of prosthetic heart valves (PHV) are not uncommon. The diagnosis is commonly established by increased gradients across PHV during transthoracic echo and restricted leaflet movement during transesophageal echo or fluoroscopy. We demonstrated both the leaflets of a thrombosed PHV,of which one was stuck in closed position by real-time three-dimensional transesophageal echocardiography. PMID- 20012206 TI - Closed-chest animal model of chronic coronary artery stenosis. Assessment with magnetic resonance imaging. AB - To evaluate the consequences of chronic non-occlusive coronary artery (CA) stenosis on myocardial function, perfusion and viability, we developed a closed chest, closed-pericardium pig model, using magnetic resonance imaging (MRI) as quantitative imaging tool. Pigs underwent a percutaneous copper-coated stent implantation in the left circumflex CA (n = 19) or sham operation (n = 5). To evaluate the occurrence of myocardial infarction, cardiac troponin I (cTnI) levels were repetitively measured. At week 6, CA stenosis severity was quantified with angiography and cine, first-pass and contrast-enhanced MRI were performed to evaluate cardiac function, perfusion and viability. In the stenting group, cTnI values significantly increased at day 3 and day 5 (P = 0.01), and normalized at day 12. At angiography, 13/19 stented pigs had a stenosis >75%. Mean degree of CA stenosis was 91 +/- 4%, range 83-98%. At contrast-enhanced MRI, mean infarct size was 7 +/- 6%, range 0.7-18.4%. Five of the 6 pigs with stenosis <75% had no infarction. Stented pigs showed significantly higher Left-ventricular volumes and normalized mass (P < 0.05), and lower ejection fraction (P = 0.03) than the sham pigs. Both wall thickening and myocardial perfusion were significantly lower in animals with at least one segment >50% infarct (23 +/- 8%; 0.05 +/- 0.01 a.u./s) and animals with only <50% infarct segments (29% +/- 12%; 0.07 +/- 0.02 a.u./s), than sham pigs (52 +/- 6%; 0.10 +/- 0.03 a.u./s) (P < 0.001; P < 0.05). This minimally-invasive animal model of chronic, non-occlusive CA stenosis, presenting a mixture of perfusion and functional impairment and a variable degree of myocardial necrosis, can be used as substitute to study chronic myocardial hypoperfusion. PMID- 20012208 TI - Growth of primary embryo cells in a microculture system. AB - We present optimal perfusion conditions for the growth of primary mouse embryonic fibroblasts (mEFs) and mouse embryonic stem cells (mESCs) using a microfluidic perfusion culture system. In an effort to balance nutrient renewal while ensuring the presence of cell secreted factors, we found that the optimal perfusion rate for culturing primary embryonic fibroblasts (mEFs) in our experimental setting is 10 nL/min with an average flow velocity 0.55 microm/s in the microchannel. Primary mEFs may have a greater dependence on cell secreted factors when compared to their immortalized counterpart 3T3 fibroblasts cultured under similar conditions. Both the seeding density and the perfusion rate are critical for the proliferation of primary cells. A week long cultivation of mEFs and mESCs using the microculture system exhibited similar morphology and viability to those grown in a petri dish. Both mEFs and mESCs were analyzed using fluorescence immunoassays to determine their proliferative status and protein expression. Our results demonstrate that a perfusion-based microculture environment is capable of supporting the highly proliferative status of pluripotent embryonic stem cells. PMID- 20012209 TI - Handheld histology-equivalent sectioning laser-scanning confocal optical microscope for interventional imaging. AB - A handheld, forward-imaging, laser-scanning confocal microscope (LSCM) demonstrating optical sectioning comparable with microtome slice thicknesses in conventional histology, targeted towards interventional imaging, is reported. Fast raster scanning (approximately 2.5 kHz line scan rate, 3.0-5.0 frames per second) was provided by a 2-axis microelectromechanical system (MEMS) scanning mirror fabricated by a method compatible with complementary metal-oxide semiconductor (CMOS) processing. Cost-effective rapid-prototyped packaging combined the MEMS mirror with micro-optical components into a probe with 18 mm outer diameter and 54 mm rigid length. ZEMAX optical design simulations indicate the ability of the handheld optical system to obtain lateral resolution of 0.31 and axial resolution of 2.85 microm. Lateral and axial resolutions are experimentally measured at 0.5 microm and 4.2 microm respectively, with field of view of 200 x 125 microm. Results of reflectance imaging of ex vivo swine liver, and fluorescence imaging of the expression of cytokeratin and mammaglobin tumor biomarkers in epithelial human breast tissue from metastatic breast cancer patients are presented. The results indicate that inexpensive, portable handheld optical microscopy tools based on silicon micromirror technologies could be important in interventional imaging, complementing existing coarse-resolution techniques to improve the efficacy of disease diagnosis, image-guided excisional microsurgery, and monitored photodynamic therapy. PMID- 20012210 TI - The challenge of validating SF-12 for its use with community-dwelling elderly in Israel. AB - PURPOSE: To evaluate the psychometric properties and feasibility of using a Hebrew version of the SF-12 for community-dwelling elderly. METHODS: Four hundred and twenty-one people aged 70+ were interviewed in a longitudinal study, using the Barthel Index, GDS and SF-12. RESULTS: Score distribution was satisfactory, with minimal missing data and item-to-item correlations were satisfactory. There were no floor and ceiling effects. Factor analysis yields the two-dimensional factors of physical and mental health and a third factor of physical role. Reliability of domain and the complete scores using the internal consistency method point to good internal consistency. Regarding construct validity of the scale, the mental health domain scores had a significant, negative relation with scores on the GDS and the physical health domain scores had a significant, positive relation with scores on the ADL scale and a significant, negative relation with scores on the IADL scale. CONCLUSIONS: The Hebrew version of the SF 12 appears to be a practical instrument for use among the elderly population and the resulting health domains correlate highly with other measures of functional, physical, and mental health used for the elderly. PMID- 20012212 TI - Organization of projections of the zona incerta of the diencephalon to pallidal structures in the dog brain. AB - Detailed studies of the organization of individual sectors in the zona incerta of the diencephalon to functionally diverse pallidal structures in the dog brain were performed by antero- and retrograde axon transport of horseradish peroxidase. The results showed that neurons in the caudal sector of the zona incerta innervate the globus pallidus and entopeduncular nucleus, which receive more extensive innervation from motor structures. The same pallidal structures receive projections from occasional neurons in the dorsal and causal sectors of the zona incerta. No connections of the zona incerta with the limbic ventral pallidum were found. PMID- 20012211 TI - Slow cooling and temperature-controlled protein crystallography. AB - In cryocrystallography, rapid sample cooling is generally deemed essential to prevent solvent crystallization and associated sample damage. We show that by carefully and completely removing all external solvent, many protein crystals can be successfully cooled to T = 100 K at only 0.1 K/s without additional penetrating cryoprotectants. Slow cooling provides an alternative when flash cooling fails, and enables diffraction studies of protein structure and function at all temperatures between T = 300 K and T = 100 K. PMID- 20012213 TI - Cytoplasmic syncytial connections between neuron bodies in the CNS of adult animals. AB - Studies of neurons in the dentate gyrus and hippocampal fields CA1 and CA2 and cerebellar granule cells were performed to test the hypothesis that there are syncytial connections between the bodies of neurons in adult higher vertebrates. Electron microscopic investigations showed that these cells were densely packed and had incomplete glial coatings. The outer cell membranes of these cells were found to be in contact, and membrane contacts in the form of tight junctions and gap junctions were seen. These areas showed membrane perforation and the establishment of syncytial connections between neurons, with all the expected ultrastructural characteristics. These connections could form between several contacting neurons, resulting in a unified functional cell cluster. These studies support the hypothesis that cytoplasmic syncytial interneuronal connections, along with synaptic and contact-type electrical connections, form not only in tissue cultures and the autonomic nervous system during early postnatal ontogenesis, but also in the CNS in adult vertebrates. PMID- 20012214 TI - Development of rat stellate ganglion neurons containing membrane-bound muscarinic receptors and purinoreceptors. AB - The aim of the study reported here was to investigate the locations and morphometric characteristics of neurons in the stellate ganglion (SG) containing muscarinic cholinoreceptors and purinoreceptors in rat pups of different ages (neonatal, 10, 20, 30, 60, and 180 days) using immunohistochemical methods. The results showed that in all animals, most neurons were immunoreactive to M1 cholinoreceptors and P2X2 and P2X6 purinoreceptors from birth. Sections showed isolated neurons containing P2X3 purinoreceptors, the numbers of which increased from day 10 of life to day 20 and then decreased again. During ontogenesis, the proportions of neurons expressing M1 cholinoreceptors and P2X2 and P2X6 purinoreceptors did not change significantly. Thus, by the moment of birth, muscarinic synaptic transmission already occurred in the SG in rats, while the final set of purinoreceptors in the neurons of this sympathetic ganglion formed by age 30 days. PMID- 20012215 TI - Morphogenesis of the paleoamygdala during the early juvenile period in rats. AB - Structural rearrangements in the paleoamygdala during the early juvenile period of development were characterized in studies on 40 Wistar rats of both sexes on days 21, 24, 28, and 31 of postnatal life. Serial frontal sections of the brain stained with cresyl violet by the Nissl method were used to study the cytoarchitectonics of structures within the paleoamygdala (the dorsomedial (Med), the posterior medial (Mep), and posterior cortical (Cop) nuclei) and the dynamics of their formation by recording of planimetric characteristics, with measurements of the numbers of neurons and gliocytes and calculation of glial and apoptotic indexes. The results obtained from these experiments provided evidence for heterochronous morphogenesis and sex-related differences in the differentiation of paleoamygdalar structures. Only the Med was differentiated on day 21 (the beginning of the early juvenile period), while the Mep could be recognized on day 24 and the Cop acquired its characteristic cytoarchitectonics only on day 31 (the beginning of the late juvenile period). Significant gender-related differences in structure were seen in the Med from day 28 and in the Cop and Mep on day 31. PMID- 20012216 TI - Location of neurons in the central nucleus of the amygdaloid body projecting to the paraventricular nucleus of the hypothalamus. AB - Experiments were performed on rats using retrograde axonal transport of horseradish peroxidase with the aim of identifying cells in the individual structures of the central nucleus of the amygdaloid body (CNAB) innervating the paraventricular nucleus of the hypothalamus. Labeled cells were seen in the ipsilateral CNAB throughout its rostrocaudal extent. The largest number of labeled neurons was seen in the middle third of the nucleus in the area corresponding to the intermediate subnucleus of the CNAB. Occasional cells were located in the medial and lateral subnuclei of the CNAB. Labeled neurons in the intermediate subnucleus were oval cells or cells of indeterminate shape. PMID- 20012217 TI - Adenoviral oncolytic suicide gene therapy for a peritoneal dissemination model of gastric cancer in mice. AB - BACKGROUND: Peritoneal dissemination of gastric cancer is often refractory to systemic therapies. Although adenoviral gene therapy has been reported to be a potentially useful therapeutic modality, the adenovirus itself has a dose limiting toxicity. A novel system was constructed using adenoviral oncolytic suicide gene therapy targeting carcinoembryonic antigen (CEA), and its therapeutic effect and the possibility to reduce the total viral dose while still preserving the antitumor effect were assessed. METHODS: Three types of adenoviruses were prepared for this novel system: (A) Ad/CEA-Cre, (B) Ad/lox CD::UPRT for a Cre/loxP system, and (C) Ad/CEA-E1 for conditionally replicating adenovirus. The antitumor effect of the oncolytic suicide gene therapy (A + B + C) was then evaluated in vitro. Mice bearing peritoneal dissemination of human gastric cancer were treated with either this system (A + B + C) or with a tenfold viral dose of suicide gene therapy (A + B). The adverse effects in terms of hepatotoxicity were then evaluated between the two groups. RESULTS: The current system (A + B + C) demonstrated significantly better cytotoxic effect for CEA producing cell lines than did suicide gene therapy (A + B) at the same viral dose in vitro. The effect of oncolytic suicide gene therapy was almost equal to that of the tenfold viral dose of suicide gene therapy in vivo. The hepatotoxicity of the two treated groups was also found to be equivalent. CONCLUSION: It was possible to reduce the total adenoviral dose of oncolytic suicide gene therapy while still preserving the antitumor effect. PMID- 20012220 TI - A comparison between time domain and spectral imaging systems for imaging quantum dots in small living animals. AB - PURPOSE: We quantified the performance of time-domain imaging (TDI) and spectral imaging (SI) for fluorescence imaging of quantum dots (QDs) in three distinct imaging instruments: eXplore Optix (TDI, Advanced Research Technologies Inc.), Maestro (SI, CRi Inc.), and IVIS-Spectrum (SI, Caliper Life Sciences Inc.). PROCEDURE: The instruments were compared for their sensitivity in phantoms and living mice, multiplexing capabilities (ability to resolve the signal of one QD type in the presence of another), and the dependence of contrast and spatial resolution as a function of depth. RESULTS: In phantoms, eXplore Optix had an order of magnitude better sensitivity compared to the SI systems, detecting QD concentrations of ~40 pM in vitro. Maestro was the best instrument for multiplexing QDs. Reduction of contrast and resolution as a function of depth was smallest with eXplore Optix for depth of 2-6 mm, while other depths gave comparable results in all systems. Sensitivity experiments in living mice showed that the eXplore Optix and Maestro systems outperformed the IVIS-Spectrum. CONCLUSION: TDI was found to be an order of magnitude more sensitive than SI at the expense of speed and very limited multiplexing capabilities. For deep tissue QD imaging, TDI is most applicable for depths between 2 and 6 mm, as its contrast and resolution degrade the least at these depths. PMID- 20012221 TI - Assessment of low-level metal contamination using the Mediterranean mussel gills as the indicator tissue. AB - PURPOSE: The aim of this study was to compare the level of metal contamination in two bays in the middle part of the Eastern Adriatic coastal zone in Croatia using the gills of mussels Mytilus galloprovincialis as indicator tissue. Despite the existing sources of contamination, previous studies with caged mussels only indicated moderate metal contamination of the Kastela Bay, contrary to the Trogir Bay in which marina and shipyard present a probable source of Cu- and Zn contamination. METHODS: The measurements of metallothioneins (MTs) and metals that induce MT synthesis (Cu, Zn, and Cd) were performed in the heat-treated gill cytosol and total proteins (TPs) in the untreated gill cytosol. MTs were determined by differential pulse voltammetry, Cu and Zn by flame atomic absorption spectrometry (AAS), Cd by graphite furnace AAS, and TPs by Bradford spectrophotometric procedure. RESULTS: The results collected in four sampling campaigns (autumn periods from 2001 to 2004) indicated that MT levels in mussel gills (expressed on dry mass basis 2.3+/-0.3 mg g(-1)) were comparable with basal levels reported in the literature (2.5+/-0.8 mg g(-1)). Observed interindividual, temporal, and spatial MT variability could be associated with different confounding factors, such as the time of sampling, total protein concentration, and mussel size rather than cytosolic levels of Cu and Zn. Metal levels, expressed on wet mass basis, in the heat-treated gill cytosol ranged from 1.33 to 11.31 microg g(-1) for Zn, from 0.72 to 2.96 microg g(-1) for Cu, and from 0.036 to 0.100 microg g(-1) for Cd. The highest Zn level was measured at Vranjic (Kastela Bay)-the site influenced by untreated domestic wastewater, while somewhat increased Zn and the highest Cu levels were found at marina and shipyard locations (Trogir Bay). The highest Cd level was measured at Inavinil (Kastela Bay). CONCLUSIONS: The observed association of gill MT levels with several biotic and abiotic factors limits its use as the biomarker of low-level metal exposure. Therefore, the use of the metal concentrations in the heat-treated gill cytosol of Mediterranean mussels should be considered for the assessment of the low-level metal contamination of coastal marine areas. PMID- 20012222 TI - A new multicompartmental reaction-diffusion modeling method links transient membrane attachment of E. coli MinE to E-ring formation. AB - Many important cellular processes are regulated by reaction-diffusion (RD) of molecules that takes place both in the cytoplasm and on the membrane. To model and analyze such multicompartmental processes, we developed a lattice-based Monte Carlo method, Spatiocyte that supports RD in volume and surface compartments at single molecule resolution. Stochasticity in RD and the excluded volume effect brought by intracellular molecular crowding, both of which can significantly affect RD and thus, cellular processes, are also supported. We verified the method by comparing simulation results of diffusion, irreversible and reversible reactions with the predicted analytical and best available numerical solutions. Moreover, to directly compare the localization patterns of molecules in fluorescence microscopy images with simulation, we devised a visualization method that mimics the microphotography process by showing the trajectory of simulated molecules averaged according to the camera exposure time. In the rod-shaped bacterium Escherichia coli, the division site is suppressed at the cell poles by periodic pole-to-pole oscillations of the Min proteins (MinC, MinD and MinE) arising from carefully orchestrated RD in both cytoplasm and membrane compartments. Using Spatiocyte we could model and reproduce the in vivo MinDE localization dynamics by accounting for the previously reported properties of MinE. Our results suggest that the MinE ring, which is essential in preventing polar septation, is largely composed of MinE that is transiently attached to the membrane independently after recruited by MinD. Overall, Spatiocyte allows simulation and visualization of complex spatial and reaction-diffusion mediated cellular processes in volumes and surfaces. As we showed, it can potentially provide mechanistic insights otherwise difficult to obtain experimentally. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1007/s11693-009-9047-2) contains supplementary material, which is available to authorized users. PMID- 20012223 TI - The consumption of food products from linseed-fed animals maintains erythrocyte omega-3 fatty acids in obese humans. AB - Based on mechanistic and epidemiological data, we raise the question of the relationship between qualitative dietary polyunsaturated fatty acids (PUFA) changes and increase in obesity. In this double-blind trial, we studied the effects on 160 overweight volunteers (body mass index, BMI >30) of a 90 days experimental diet rich principally in animal fat with a low PUFA/saturated fatty acid (SFA) ratio but a low n-6/n-3 ratio, using animal products obtained from linseed-fed animals. The control diet provided less animal fat, a higher PUFA/SFA ratio and a higher n-6/n-3 ratio. Both diets excluded seafood. In the experimental group, we observed a significant increase in red blood cell (RBC) alpha-linolenic acid content and a slight increase in EPA and DHA derivatives, while in the control group we observed a significant reduction in EPA and DHA content. Between groups now, the difference in the three n-3 fatty acids changes in RBC was significant. This demonstrates that plasma EPA and DHA levels can be maintained without fish if products from linseed-fed animals are used. During the diets, we noted a significant reduction in weight, BMI and hip circumference within both groups of volunteers. However, no significant difference was observed between the control group and the experimental group. Interestingly, 150 days after the end of the trial (i.e., day 240), we noted a significant weight gain in the control group, whereas no significant weight gain was observed in the experimental group. This was also observed for the BMI and hip circumference. Moreover, significant differences in BMI (P < 0.05) and weight (P = 0.05) appeared between the two groups, showing in both cases a smaller increase in the experimental group. During the 90 days trial, we did not observe any differences between groups in terms of total cholesterol, HDL cholesterol, LDL cholesterol or triglycerides, suggesting that the saturate content and the P/S ratio are not as important as the n-6 and n-3 fatty acid composition. PMID- 20012224 TI - Bat-associated leptospirosis. AB - Leptospirosis is a globally prevalent disease that affects humans, causing systemic illness that may lead to multi-organ involvement. Clinical signs include sudden fever, general malaise, muscular pain, conjunctival suffusion, and jaundice. Disease is caused by pathogenic bacteria including over 200 serologic variants. Most serologic variants have primary reservoirs in wild mammals, which continually infect and colonize domesticated animals. The organism has been recovered from rats, swine, dogs, cattle, and other animals, notably bats. Most studies have focused on domestic animals as reservoir hosts; however, because of their abundance, spatial distribution, and interrelationship with domestic animals, bats are becoming an epidemiologically significant source of leptospires. We present a case of serologically confirmed leptospirosis after bat exposure to add to the growing literature of bats as a possible source of transmission. Recognition of the common presentation of leptospirosis and Weil's disease, and identification of animal vectors, including bats, allows for the selection of appropriate antibiotic management to aid in resolution of symptomotology. PMID- 20012225 TI - Regenerative medicine, tissue engineering and vascular surgery: twenty first century clinical challenges. PMID- 20012226 TI - Laparoscopic gastric bypass as a revision procedure after transoral gastroplasty. AB - BACKGROUND: Transoral gastroplasty (TOGA) has been offered as an investigational alternative restrictive procedure in our hospital for the last 3 years. Since laparoscopic Roux-en-Y gastric bypass (LRYGBP) can be performed as a revisional surgery after failure of a restrictive surgery, this study reports on the feasibility of conversion of TOGA into a LRYGBP in case of failure of the endoscopic procedure. METHODS: Since 2006, 71 TOGA procedures were performed in morbidly obese patients. Four patients underwent an LRYGBP after TOGA procedure for unsatisfactory results after 1-year observation. All of them had undergone a second procedure in which additional TOGA restrictions were placed to attempt to tighten the pouch before being referred for LRYGBP. The surgical outcome of these patients was analysed. RESULTS: All four patients were easily converted to a LRYGBP with no major complication and no mortality. The operative results (operating time, morbidities, follow-up) of all LRYGBP post TOGA were similar to primary LRYGBP performed by the same operator. CONCLUSION: LRYGBP post-TOGA apparently can be done without any trouble. The performance of TOGA does not seem to interfere with the short-term results of the LRYGBP. PMID- 20012227 TI - IL-15 receptor deletion results in circadian changes of locomotor and metabolic activity. AB - Interleukin-15 (IL-15) is a cytokine produced in the normal brain that acts on its specific receptor IL-15Ralpha and co-receptors IL-2Rbeta and IL-2Rgamma in neuronal cells. The functions of the cerebral IL-15 system, however, are not yet clear. To test the hypothesis that IL-15Ralpha regulates metabolic activity and body temperature, we quantified the specific metabolic phenotype of IL-15Ralpha knockout mice. These normal-appearing mice were leaner with lower fat composition. During the entire circadian cycle, the knockout mice had a significantly higher acrophase in locomotor activity and heat dissipation. During the light phase, there was significantly greater food intake, oxygen consumption, and carbon dioxide production. The difference in the dark and light phases suggests that IL-15Ralpha participates in circadian rhythm regulation. The higher oxygen consumption in the light phase indicates adaptive thermogenesis in the knockout mice. The body temperature of the receptor knockout mice was significantly higher than the control in the light phase, and this was mainly caused by a large difference occurring between 0600 and 0900 h. In addition to the metabolic chamber studies and circadian rhythm analyses, qPCR of hypothalamic homogenates indicated higher mRNA expression of orexin and transient receptor potential vanilloid 4 cation channels. Consistent with a direct role of IL 15Ralpha in the hypothalamus, IL-15 treatment of the wild-type mice induced c-Fos expression in the preoptic area. We conclude that activation of hypothalamic neurons by IL-15 in mice contributes to thermoregulation and modifies the metabolic phenotype. PMID- 20012228 TI - Nerve growth factor-induced protection of brain capillary endothelial cells exposed to oxygen-glucose deprivation involves attenuation of Erk phosphorylation. AB - Nerve growth factor (NGF) was recently characterized as an angiogenic factor inducing proliferation, migration, and capillary sprouting in endothelial cells (ECs) of different vascular beds. While NGF neuroprotective effects on neurons were described, its survival-inducing effects on brain capillary ECs were not yet addressed. Using a model of oxygen-glucose deprivation (OGD) followed by reoxygenation, we demonstrated that NGF conferred protection in brain capillary ECs. These cells express TrkA and p75(NTR) receptors and respond to NGF by stimulation of Erk1/2 phosphorylation and stimulation of proliferation and migration. The NGF protective effect was dose-dependent, inhibited by NGF/TrkA antagonist, K252a, and required presence of NGF during both OGD and reoxygenation phases while the major protective effect was related to decreased cell death during the reoxygenation phase. A causal relationship was found between NGF induced protection and attenuation of OGD-induced Erk1/2 phosphorylation, supporting the death-promoting role of insult-induced Erk1/2 phosphorylation in the brain capillary ECs. These results emphasize the importance of NGF in the process of EC survival in response to ischemic injury and suggest fine-tuning regulation of Erk1/2 phosphorylation, extending the neuroprotective impact of NGF from sympathetic neuroendocrine cells to brain capillary ECs as the other element in the neurovascular tandem. PMID- 20012229 TI - Pedunculated early ampullary carcinoma treated by ampullectomy: report of a case. AB - CASE REPORT: A 30-year-old woman was admitted to our hospital for treatment of an ampullary tumor. Upper gastrointestinal endoscopy revealed a pedunculated tumor in the ampulla of Vater with a diameter of 50 mm, which was biopsied and diagnosed as tubulovillous adenoma with moderate atypia. Endoscopic ultrasonography demonstrated a hypoechoic tumor limited to the mucosa and without evidence of lymph node metastasis. Since endoscopic resection was not indicated because of the large size and pedunculated morphology with a long stalk, the patient underwent ampullectomy and papilloplasty. Histological examination revealed well-differentiated tubular adenocarcinoma in tubular adenoma with severe atypia limited to the mucosa. The patient remains well with no evidence of recurrence 10 months after resection. DISCUSSION: Ampullectomy is an established method for ampullary tumor, but such a tumor with a long stalk is rare. PMID- 20012230 TI - Stem cell therapy improves the outcome of liver resection in cirrhotics. AB - INTRODUCTION: Significant proportions of liver cirrhotic patients develop hepatocellular carcinoma and have to undergo hepatic resection. The compromised cirrhotic liver cannot withstand further removal of hepatic tissue, thus, leading to postoperative complication and death. METHODS: In this study, we enrolled 20 patients having liver cirrhosis with hepatocellular carcinoma and randomly assigned them into two groups to receive autologous stem cells or placebo. RESULTS: After 3 weeks, all participants underwent liver resection and were followed for 12 weeks postoperative. We observed that the group receiving preoperative stem cell therapy had shown a significant improvement in all parameters of liver function and had no postoperative complications compared to the group treated with placebo, which showed no improvement in liver parameters and had postoperative complications. DISCUSSION: In conclusion, autologous stem cell therapy can improve the surgical outcome in cirrhotic livers and should be considered as an adjuvant treatment in such patients undergoing hepatic resection. PMID- 20012232 TI - The postthrombotic syndrome. AB - Following deep vein thrombosis (DVT), one of every two patients will develop postthrombotic syndrome (PTS), which causes remarkable consequences on the socioeconomic level. Residual thrombosis is an important predictor of PTS, and severe early symptoms, old age, obesity, improper anticoagulation, recurrent thrombosis and varicose veins are major risk factors. Diagnosis of PTS is mainly based on the clinical findings for patients with a history of DVT, while in those without it, instrumental diagnosis might help in detecting a previous DVT. Prompt administration of adequate compression elastic stockings (ECS) in patients with symptomatic DVT reduces the frequency of PTS by half. Usually, the management of an established PTS is demanding, and often discouraging. However, when carefully supervised and instructed to wear proper ECS, more than 50% of patients either remain quiescent or improve during long-term follow-up. PMID- 20012233 TI - TGFB1 509 C/T polymorphism and colorectal cancer risk: a meta-analysis. AB - Transforming growth factor beta (TGF-beta) is a cytokine. The TGF-beta signaling pathway plays an important role in controlling cell proliferation and differentiation involved in colorectal carcinogenesis. In mammalian cells, TGFB1 is the most abundant subtype of TGF-beta. The 509 C/T polymorphism in TGFB1 has been implicated in colorectal cancer risk. However, published data remain conflicting. To derive a more precise estimation of the relationship, a meta analysis of 994 cases and 2,335 controls from five published case-control studies was performed. Overall, significantly increased colorectal cancer risks were found for CC versus TT (OR=1.62; 95% CI: 1.30-2.02; Pheterogeneity=0.118), TC+CC versus TT (OR=1.30; 95% CI: 1.08-1.58; Pheterogeneity=0.259) and CC versus TC+TT (OR=1.48; 95% CI: 1.26-1.75; Pheterogeneity=0.244). In the subgroup analysis by ethnicity, significantly increased risks were also found among Asians for CC versus TT (OR=1.77; 95% CI: 1.40-2.24; Pheterogeneity=0.519), TC+CC versus TT (OR=1.38; 95% CI: 1.13-1.68; Pheterogeneity=0.679) and CC versus TC+TT (OR=1.58; 95% CI: 1.31-1.89; Pheterogeneity=0.340). However, no significant associations were found among Europeans for all genetic models. This meta-analysis showed that TGFB1 509 C allele is a risk factor for developing colorectal cancer in Asians. PMID- 20012231 TI - Pregnancy outcomes in patients with autoimmune diseases and anti-Ro/SSA antibodies. AB - Anti-Ro/SSA antibodies are associated with neonatal lupus (congenital heart block (CHB), neonatal transient skin rash, hematological and hepatic abnormalities), but do not negatively affects other gestational outcomes, and the general outcome of these pregnancies is now good, when followed by experienced multidisciplinary teams. The prevalence of CHB, defined as an atrioventricular block diagnosed in utero, at birth, or within the neonatal period (0-27 days after birth), in the offspring of an anti-Ro/SSA-positive women is 1-2%, of neonatal lupus rash around 10-20%, while laboratory abnormalities in asymptomatic babies can be detected in up to 27% of cases. The risk of recurrence of CHB is ten times higher. Most of the mothers are asymptomatic at delivery and are identified only by the birth of an affected child. Half of these asymptomatic women develop symptoms of a rheumatic disease, most commonly arthralgias and xerophtalmia, but few develop lupus nephritis. A standard therapy for CHB is still matter of investigation, although fluorinated corticosteroids have been reported to be effective for associated cardiomyopathy. Serial echocardiograms and obstetric sonograms, performed at least every 1-2 weeks starting from the 16th week of gestational age, are recommended in anti-Ro/SSA-positive pregnant women to detect early fetal abnormalities that might be a target of preventive therapy. PMID- 20012234 TI - Comparative proteomic profiling identified sorcin being associated with gemcitabine resistance in non-small cell lung cancer. AB - Although gemcitabine-based chemotherapy is one of the more effective chemotherapy regimens against NSCLC, there are still many patients who do not benefit from this therapy. The mechanism of initial or acquired resistance to gemcitabine chemotherapy remains unknown. In this study, we investigated the protein profiling in gemcitabine-resistant and gemcitabine-sensitive NSCLC cell lines by a proteomic technology in order to identify novel gemcitabine resistance associated biomarkers for NSCLC patients. The proteomic profiling of NSCLC cell line H460 and its gemcitabine-resistant subline H460/GEM were compared by an isotope-coded affinity tag technology and tandem mass spectrometry. We further validated the expression of sorcin, a gemcitabine-resistance-related protein identified by proteomics, in 62 NSCLC specimens by immunohistochemistry. Fourteen gemcitabine resistance-related proteins were identified including nine up regulated proteins and five down-regulated proteins. Immunohistochemical results demonstrated that sorcin staining was seen in 66.1% of NSCLC tumors, and sorcin overexpression was associated with gemcitabine resistance and a poor prognosis in NSCLC patients. In conclusion, sorcin might play an important role in the resistance to gemcitabine, and it could also be a novel candidate biomarker for predicting the response of NSCLC patients to gemcitabine treatment. PMID- 20012235 TI - Probable sorafenib-induced reversible encephalopathy in a patient with hepatocellular carcinoma. PMID- 20012236 TI - Intramedullary nailing as a 'second hit' phenomenon in experimental research: lessons learned and future directions. AB - BACKGROUND: The 'second hit' phenomenon is based on the fundamental concept that sequential insults, which are individually innocuous, can lead to overwhelming physiologic reactions. This response can be expressed in several organic systems and can be examined by measurement of several parameters. PURPOSES: The purpose of this study was to evaluate the incidence of systemic effects of intramedullary nailing and the role of concurrent head and thoracic injuries as they have been recorded in vivo. We also wanted to determine what would be the optimal animal model for future research and what variables should be investigated. METHODS: We reviewed the available literature of animal studies that used surgery, and particularly nailing, as a second hit. The reviewed studies were retrieved through an electronic search of the MEDLINE database. We analyzed the methods of creating the first and second hits (nailing), the characteristics of the animal models, the variables examined, and the pathophysiologic responses, which appeared after the second hit. RESULTS: Second hit reamed intramedullary nailing was found to provoke consumption of coagulation factors, whereas the effect of unreamed nailing on coagulation factors appears inconsistent. Hemodynamic factors were affected only transiently by the second hit, whereas the pulmonary function was affected only when the first hit included lung injury. CONCLUSIONS: Thoracic and head injuries predispose to an aggravated second hit. Primate animal models are considered to be closer to clinical reality and should be preferred for future studies. Future studies should include measurements of proinflammatory and antiinflammatory markers. PMID- 20012237 TI - Functional performance with a single-radius femoral design total knee arthroplasty. AB - BACKGROUND: Better muscular recovery of the extensor mechanism after TKA is claimed by femoral designs based on a sagittal single radius. QUESTIONS/PURPOSES: We aimed to compare postoperative knee performance through the Knee Society scores, flexor and extensor muscle function, stability, and gait of a series of patients receiving a posterior stabilized, cemented TKA, with a single-radius femoral design. METHODS: We compared a series of patients treated with a single radius femoral design TKA to a simultaneous series of patients receiving a multiradius femoral design. Both groups were similar in demographics and preoperative Knee Society scores. The clinical pathways were identical. Outcome assessment included Knee Society scores, isokinetic assessment, stabilometry, and gait cycle analysis. RESULTS: We observed higher functional Knee Society scores (86.6 +/- 1.89 versus 80.3 +/- 1.90), fewer physiotherapy sessions (19.9 +/- 4.65 versus 22.2 +/- 3.34), and less time with two crutches (3.5 +/- 1.2 versus 5.2 +/ 1.04 weeks) for patients receiving the single-radius design. Isokinetic evaluation showed decreased flexion peak torque (40.3 +/- 7.9 versus 48.7 +/- 9.6), increased extension peak torque (77.2 +/- 16.1 versus 69.1 +/- 14.4), and lower flexor/extensor ratio (0.5 +/- 0.08 versus 0.7 +/- 0.1) in patients with the single-radius design. Stabilometry showing less relative oscillation, and gait cycle indirectly confirmed better support in the limb with the single-radius design. CONCLUSIONS: The studied single-radius femoral design showed better functional short-term outcome and better extensor performance. LEVEL OF EVIDENCE: Level III, therapeutic study. See the Guidelines for Authors for a complete description of levels of evidence. PMID- 20012238 TI - Case report: perioperative use of protein c concentrate for protein C deficiency in THA. AB - BACKGROUND: Perioperative management of patients with heterozygous protein C deficiency is challenging because of the competing risks of bleeding and recurrent thrombosis. CASE DESCRIPTION: We report the case of a 74-year-old man with protein C deficiency and heterozygous prothrombin G20210A gene mutation who had a successful left THA with perioperative administration of human zymogen protein C concentrate in addition to anticoagulation with enoxaparin. LITERATURE REVIEW: Several studies have reported the use of protein C concentrate in severe sepsis-associated purpura fulminans in patients with severe congenital protein C deficiency who have had thrombotic events. We reviewed studies and case reports pertinent to the treatment of patients with protein C deficiency, especially in the perioperative setting. We report the case of a patient undergoing THA in whom we used human zymogen protein C concentrate. PURPOSES AND CLINICAL RELEVANCE: THA, a particularly high-risk procedure, is associated with a 40% to 70% incidence of venographic deep venous thrombosis and a 2% to 3% incidence of symptomatic deep venous thrombosis. These risks are greater in people with thrombophilic defects such as protein C deficiency. The use of human zymogen protein C in our patient with heterozygous protein C deficiency during the perioperative period of a THA was associated with no evidence of excessive bleeding, hematoma, deep venous thrombosis, or pulmonary embolism. PMID- 20012239 TI - Comparing exponentially weighted moving average and run rules in process control of semiquantitative immunogenicity immunoassays. AB - The immunogenicity immunoassay validation process ensures development of a robust, reproducible method. However, no matter how well developed, validated, and maintained a method is, in the course of running a large number of samples over time, it is not uncommon to see bad reagents, poorly calibrated equipment, personnel errors, or other unknown and unpredictable factors that have an impact in the performance of the method and quality of the sample results. The immunogenicity immunoassay thus needs to be closely monitored with an internal statistical quality control process overtime to ensure a consistent and reliable output. The statistical process control has been widely applied to monitor manufacturing processes and in clinical laboratories. Its application to immunogenicity immunoassays is relatively novel. Limited guidance is available to implement the process to monitor semiquantitative immunogenicity immunoassay performance. Here, we have performed a suitability evaluation for process control charts with actual laboratory data from three immunogenicity immunoassay methods each utilizing a different technology platform. Additionally, a panel of prepared samples designed to assess long-term method performance were periodically evaluated for over a year. Finally, we make recommendations for an internal quality control process based on the results of these evaluations. PMID- 20012240 TI - Mammographic density and matrix metalloproteinases in breast tissue. AB - Mammographic density is a strong risk factor for breast cancer, yet the underlying histopathologic correlates are not clear. Matrix metalloproteinases (MMPs) and their inhibitors (TIMPs) play important roles in multiple stages of tumorigenesis. This study examined the association between mammographic density and expression of MMPs 1, 3, 9, and 12 and TIMP3 in benign and malignant breast tissue of 277 women with mainly Caucasian and Japanese ancestry. Tissue microarrays with up to 4 benign and 4 malignant cores per woman were stained immunohistochemically and evaluated. Digitized prediagnostic mammograms were assessed for densities using a computer-assisted method. General linear models adjusted for known confounders were applied to estimate mean densities by staining category. Strong expression of all MMPs was about twice as frequent in malignant as in benign tissue, while TIMP3 expression in stromal tissue was higher in benign than malignant cores. For MMP3 and 9, less than 10% of cores stained positive; thus, they were not further analyzed. None of the markers showed a statistically significant association with breast density in the entire study population and ethnic-specific results were conflicting and difficult to explain. Although not statistically significant, mean density was consistently lower with more extensive TIMP3 expression in stromal and epithelial tissue. These findings indicate that the higher breast cancer risk in women with dense breasts may be influenced by lower TIMP3 expression. However, future investigations into activities and ratios of additional proteases and their inhibitors as well as other pathways, such as inflammation, are needed. PMID- 20012241 TI - Diagnostic dilemma of a subcutaneous nodule following a trauma in a child: immunohistochemical examination put the final diagnosis of deep granuloma annulare. AB - The diagnostic dilemma is discussed on a child who presented a granulomatous process of the inferior limb following a trauma. It was originally considered epithelioid sarcoma (ES) and later diagnosed as deep granuloma annulare (DGA). In combination with the clinical assessment, the plain radiographs, the ultrasound and the magnetic resonance proved inadequate in reaching diagnosis and in excluding neoplasia. Biopsy was required, and the following immunohistochemical examination put the definitive diagnosis of deep granuloma annulare. Development of DGA after trauma seems to be rare, but it is a possible event and further observations are needed to explain the triggering roles in the etiopathogenesis of this disease. PMID- 20012242 TI - Anti-HIV drugs nevirapine and efavirenz affect anxiety-related behavior and cognitive performance in mice. AB - Nevirapine (NVP) and efavirenz (EFV) belong to the class of anti-HIV drugs called non-nucleoside reverse transcriptase inhibitors (NNRTIs), commonly used as part of highly active antiretroviral therapy (HAART). Although the HAART is able to bring down viral load to undetectable levels and restore immune function, their prolonged use causes several adverse effects. It has been demonstrated that both NVP and EFV are able to cross the blood-brain barrier, causing important central nervous system-related side effects. Thus, this study investigated the effects of chronic administration of EFV (10 mg/kg) and NVP (3.3 mg/kg) in mice submitted to two distinct series of experiments, which aimed to evaluate: (1) the emotional behavior (elevated plus-maze, forced swimming, and open-field test) and (2) the cognitive performance (object recognition and inhibitory avoidance test) of mice. Our results demonstrated that EFV, but not NVP, reduced the exploration to open arms in the elevated plus-maze test. Neither NVP nor EFV altered mouse behavior in the forced swimming and open-field tests. Both drugs reduced the recognition index in the object recognition test, but only EFV significantly impaired the aversive memory assessed in the inhibitory avoidance test 24 h after training. In conclusion, our findings point to a genuine anxiogenic-like effect to EFV, since it reduced exploration to open arms of elevated plus-maze test without affecting spontaneous locomotion. Additionally, both drugs impaired recognition memory, while only the treatment with EFV impaired significantly aversive memory. PMID- 20012243 TI - [Emergency room management of severely injured patients]. AB - BACKGROUND: In cases involving major trauma life-threatening situations should be immediately diagnosed and treated. Clinical algorithms can potentially decrease the rate of complications and errors. The purpose of this study was to investigate the incidence of deviations from a multislice computed tomography based trauma room algorithm. MATERIALS AND METHODS: During a primary trauma survey an independent study monitor observed the on site treatment sequence step by step. Time intervals between admission and start of each procedure were recorded. Deviations from the algorithm and delays were analyzed. RESULTS: In 57 trauma patients a total of 49 deviations were documented. Median time between admission and transfer to the adjacent MSCT room was 9 min. Of the patients 11 were bypassed to the MSCT suite without a primary survey (19.3%). In 2 cases an absence of non-invasive blood pressure monitoring was recorded (3.5%) and 3 patients with potential cervical spine trauma were not immobilized at the scene or during primary survey (5.3%). In 8 cases focused assessment with sonography for trauma (FAST) was not performed (14%). Contrary to the algorithm 10 patients received an arterial or central venous line during initial treatment (18%) resulting in a median delay of 8 min. The deviations from the algorithm resulted in no adverse effects on complications or mortality. CONCLUSION: Self-critical analysis of trauma resuscitation can increase the quality of treatment by revealing constantly recurring faults. PMID- 20012244 TI - [Intratracheal foreign body caused by a sheared endotracheal intubation stylet]. AB - After orotracheal intubation of a 30-year-old man with a flexible tube, a floating foreign body was noticed inside the tube. The subsequent bronchoscopy revealed a second foreign body near the bifurcation of the trachea. Both foreign bodies were removed successfully and identified as parts of the plastic-sheathed stylet. Fatigue of material is discussed as the reason for the broken reusable stylet during intubation. PMID- 20012245 TI - [Intra-operative drop in sevoflurane and oxygen concentrations. Leakage of the ventilation system]. AB - After problem-free induction of narcosis in an 84-year-old female patient an intra-operative drop in sevoflurane and oxygen concentrations occurred during low flow anesthesia. Although the concentrations of sevoflurane and oxygen in the fresh gas flow were increased no adequate elevation of the inspiratory concentrations could be achieved. Disconnection of the Drager Primus IE manual bag-valve-mask could be identified as the cause of the drop in concentrations. Interestingly no error alarm function was initiated. This case demonstrates how important knowledge of the function, set-up and alarm conditions of respiratory machines is. This should be an important component of training in anesthesiology as well as securely established algorithms for difficult ventilation to ensure safe anesthesia despite technical failures. PMID- 20012246 TI - [Paediatric simulation today and tomorrow. Perspectives and concepts]. AB - The confrontation with critically ill newborns, infants and small children is rare and poses a particular challenge for the medical team. Confident technical and non-technical skills are essential for successful emergency treatment. Paediatric simulators facilitate a didactic infrastructure, linking textbook theory with experience-based practice. To summarize the current status of paediatric simulation in Germany, Austria and Switzerland an online survey of all associated centres was conducted. Paediatric simulation is currently available at 24 centres, which have 39 paediatric simulators available, including 8 for newborns, 26 for infants and 5 for children. A certain congruence of standards is detectable among these centres and most instructors have completed a specialized instructor training. Of the instructors 26% are specialized nursing personnel and 67% are physicians of which most are paediatricians and anaesthesiologists. Many centres (38%) operate solely by means of the enthusiastic dedication of the employees who organize various activities during their free time. Nearly all centres (92%) place particular emphasis on non-technical skills which include the interpersonal aspects of crisis resource management. Video-supported debriefing is considered to be the basis for effective training. Within the scope of the recently established PaedSim project the curricula of paediatric simulation courses should be more structured and internationally standardized, thereby increasing both efficacy and sustainability of these training programs. PMID- 20012247 TI - [Sponsoring of medical conferences, workshops and symposia by pharmaceutical companies. Physicians must be wary of this!]. AB - The longstanding conventional forms of cooperation between medical organizations and physicians on the one hand and the pharmaceutical industry and manufacturers of medical products on the other hand nowadays hold the risk of coming into conflict with the public prosecutor. Typical circumstances which are taken up by the investigating authorities are financial supports of medical conferences, workshops and symposia. To understand the problem under criminal law it is important to become acquainted with the protective aim of the statutory offences of the acceptance of benefits according to section sign 331 of the Penal Code (Strafgesetzbuch, StGB) and of corruption according to section sign 332 of the Penal Code. The "trust of the general public in the objectivity of governmental decisions" must be protected and the "evil appearance of the corruptibility of official acts" must be counteracted. A basic differentiation is made between physicians with and without office-bearing functions. By paying attention to the recommendations and basic principles of cooperation between the medical profession and the healthcare industry presented in this article (transparency principle, equivalence principle, documentation principle and separation principle) the emergence of any suspicious factors can be effectively avoided. PMID- 20012251 TI - Regurgitation of my free lunch: the clinical toxicology and pharmaceutical company relationship. PMID- 20012252 TI - In response to Morris et al.'s "Neuroleptic malignant syndrome developing after acute overdose with olanzapine and chlorpromazine". PMID- 20012253 TI - In response to Barbee et al.'s "Analysis of mushroom exposures in Texas requiring hospitalization, 2005-2006". PMID- 20012254 TI - Fewer grammatical errors means less editing. PMID- 20012255 TI - [Diagnostic pitfalls with Cushing's syndrome]. AB - Adrenal cortical carcinoma is a rare diagnosis and may present with hormone secretion. A histological differentiation between an adrenal cortical adenoma and carcinoma can be very difficult. However, a fast diagnosis including staging and complete surgical resection is pivotal for the prognosis of an adrenal cortical carcinoma. Metastasing adrenal cortical carcinoma should be treated with a mitotane based chemotherapy, and inclusion in the "firm-act study" is highly recommended. The present case report demonstrates the diagnostic pitfalls in a female patients with Cushing's syndrome who suffered from metastasing adrenal cortical carcinoma. PMID- 20012256 TI - [Von Willebrand disease : diagnosis and management]. AB - Von Willebrand's disease is an inherited bleeding disorder with a prevalence as high as 1% in the general population. The disease is caused by the quantitative deficiency or dysfunction of von Willebrand factor (VWF), a large multimeric glycoprotein. VWF has two main functions in hemostasis: it is essential for platelet-plug formation as an adhesion protein and it forms a non-covalent complex with coagulation factor VIII in plasma, thereby protecting it from inactivation and clearance. Inherited Von Willebrand's disease has been subdivided into 3 categories that reflect pathophysiology: partial quantitative deficiency of VWF (Type 1), qualitative deficiency (Type 2) and total deficiency (Type 3). The major clinical hallmark in Von Willebrand's disease is an increased tendency to mucocutaneous bleeding. Increased bleeding may also occur in sites such as muscles and joints when the level of factor VIII is particularly low. The mainstays of therapy are desmopressin, which induces secretion of autologous factor VIII and VWF into plasma, and plasma concentrates, which supply allogenic forms of these moieties. Other forms of treatment can be considered as adjunctive to these. PMID- 20012257 TI - [Dysphagia, heartburn and esophageal bolus impaction in a 43-year old male patient: not always reflux disease]. AB - We report a case of a 43 year old male patient, who was admitted with recurring esophageal bolus impactions. Since his childhood he has been complaining about dysphagia and was unable to swallow medication. He also complained about heartburn. The last esophageal bolus impaction was some weeks ago. After elimination of the bolus impaction with a rigid endoscope we found a high grade stenosis in the proximal esophagus that could not even be passed with a children's endoscope. An initial treatment of eosinophil esophagitis would be the therapy with a local corticoid for 6-9 month. In patients with typical rings or stenosis a dilation therapy might be necessary. PMID- 20012258 TI - Spider fibers and the apparent fungal colonization of rock-art caves. PMID- 20012259 TI - [Diagnosis and therapy of GIST]. PMID- 20012261 TI - Fine mapping of a gene for non-pollen type thermosensitive genic male sterility in rice (Oryza sativa L.). AB - The thermo-sensitive genic male sterility (TGMS) lines play a crucial role in two line hybrid rice production. For a practical TGMS line, the stability of male sterility is one of the most important technical indicators. In this study, XianS, a spontaneous mutant with stable male sterility from an indica rice cultivar Xianhuangzhan, was classified as a non-pollen type TGMS line. The critical non-pollen sterility point temperature of XianS was determined as 27 degrees C. Genetic analysis demonstrated that the non-pollen sterility in XianS was controlled by a single recessive gene. Using SSR markers and bulked segregant analysis, the TGMS gene in XianS was fine mapped to a 183 kb interval between RMAN81 and RMX21 on chromosome 2. Two markers, 4039-1 and RMX14 completely cosegregated with this gene. Allelism test indicated that the non-pollen phenotype in seven non-pollen type TGMS lines from different sources, XianS, AnnongS-1, Q523S, Q524S, N28S, G421S, and Q527S is caused by the same TGMS gene. Although the location of TGMS gene in XianS is close to the gene OsNAC6, a previously identified candidate gene of tms5 in AnnongS-1, the sequence of OsNAC6 and its promoter region was identical in TGMS line XianS, AnnongS-1, and wild type Xianhuangzhan. These results suggest that the non-pollen type TGMS trait probably be controlled by the same TGMS gene in different TGMS rice lines, but its real candidate gene still need to be further studied and identified. PMID- 20012262 TI - Validation of mega-environment universal and specific QTL associated with seed yield and agronomic traits in soybeans. AB - The value of quantitative trait loci (QTL) is dependent on the strength of association with the traits of interest, allelic diversity at the QTL and the effect of the genetic background on the expression of the QTL. A number of recent studies have identified QTL associated with traits of interest that appear to be independent of the environment but dependent on the genetic background in which they are found. Therefore, the objective of this study was to validate universal and/or mega-environment-specific seed yield QTL that have been previously reported in an independent recombinant inbred line (RIL) population derived from the cross between an elite Chinese and Canadian parent. The population was evaluated at two field environments in China and in five environments in Canada in 2005 and 2006. Of the seven markers linked to seed yield QTL reported by our group in a previous study, four were polymorphic between the two parents. No association between seed yield and QTL was observed. The result could imply that seed yield QTL were either not stable in this particular genetic background or harboured different alleles than the ones in the original mapping population. QTL(U) Satt162 was associated with several agronomic traits of which lodging was validated. Both the non-adapted and adapted parent contributed favourable alleles to the progeny. Therefore, plant introductions have been validated as a source of favourable alleles that could increase the genetic variability of the soybean germplasm pool and lead to further improvements in seed yield and other agronomic traits. PMID- 20012263 TI - Identification and analysis of QTLs controlling cold tolerance at the reproductive stage and validation of effective QTLs in cold-tolerant genotypes of rice (Oryza sativa L.). AB - Low temperature or cold stress is one of the major constraints of rice production and productivity in temperate rice-growing countries and high-altitude areas in the tropics. Even though low temperature affects the rice plant in all stages of growth, the percent seed set is damaged severely by cold and this reduces the yield potential of cultivars significantly. In this study, a new source of cold tolerant line, IR66160-121-4-4-2, was used as a donor parent with a cold sensitive cultivar, Geumobyeo, to produce 153 F(8) recombinant inbred lines (RILs) for quantitative trait locus (QTL) analysis. QTL analysis with 175 polymorphic simple sequence repeat (SSR) markers and composite interval mapping identified three main-effect QTLs (qPSST-3, qPSST-7, and qPSST-9) on chromosomes 3, 7, and 9. The SSR markers RM569, RM1377, and RM24545 were linked to the identified QTLs for cold tolerance with respect to percent seed set using cold water (18-19 degrees C) irrigation in the field and controlled air temperature (17 degrees C) in the greenhouse. The total phenotypic variation for cold tolerance contributed by the three QTLs was 27.4%. RILs with high percent seed set under cold stress were validated with linked DNA markers and by haplotype analysis that revealed the contribution of progenitor genomes from the tropical japonica cultivar Jimbrug (Javanica) and temperate japonica cultivar Shen-Nung89 366. Three QTLs contributed by the cold-tolerant parent were identified which showed additive effect on percent seed set under cold treatment. This study demonstrated the utility of a new phenotyping method as well as the identification of SSR markers associated with QTLs for selection of cold-tolerant genotypes to improve temperate rice production. PMID- 20012264 TI - Interaction of the human plasma membrane monoamine transporter (hPMAT) with antidepressants and antipsychotics. AB - Monoamine neurotransmission is efficiently terminated through synaptic reuptake of released neurotransmitters by high-affinity Na(+)- and Cl(-)-dependent neuronal monoamine transporters of the SLC6A family located in the plasma membrane of presynaptic nerve terminals. Recently, a low-affinity, high-capacity Na(+)- and Cl(-)-independent plasma membrane monoamine transporter (PMAT) belonging to the SLC29 solute carrier family has been cloned. PMAT was shown to transport monoamine neurotransmitters as well as organic cations such as 1-phenyl 4-methyl-pyridinium (MPP(+)). Thus, the PMAT which is highly expressed in the human brain may be involved in the modulation of central monoaminergic neurotransmission and it may be a target for drugs used to treat depression and schizophrenia, i.e., dysregulations of the monoamine homeostasis in the central nervous system (CNS). Therefore, we examined in transfected cells the influence on [(3)H]-MPP(+) transport by the human PMAT (hPMAT) of nine monoamine transport inhibiting antidepressants (ADs) belonging to pharmacologically diverse classes (imipramine, desipramine, amitriptyline, bupropion, fluoxetine, sertraline, paroxetine, reboxetine, and venlafaxine), of the atypical ADs tianeptine and trimipramine and of five antipsychotics (levomepromazine, haloperidol, clozapine, olanzapine, and risperidone). All examined drugs inhibited the hPMAT; however, half-maximum inhibition (IC(50)) was observed at concentrations which were much higher than reported clinical plasma concentrations of these drugs. Thus, inhibition of the hPMAT by these CNS drugs may not (or only marginally) contribute to their therapeutic effects. PMID- 20012265 TI - Prostaglandins of the E series inhibit monoamine release via EP3 receptors: proof with the competitive EP3 receptor antagonist L-826,266. AB - Prostaglandin E(2) (PGE(2)) and its analogue sulprostone inhibit noradrenaline and serotonin release in rodent tissues. We examined whether the receptor involved is blocked by the EP(3) antagonist L-826,266, whether such receptors also occur on central cholinergic neurones and retinal dopaminergic cells, whether PGE(2) is produced by the degradation of the endocannabinoid virodhamine and whether EP(3) receptor activation stimulates (35)S-GTPgammaS binding. Transmitter release was studied as electrically evoked tritium overflow in superfused tissues preincubated with (3)H-noradrenaline (which in the guinea pig retina labels dopaminergic cells), (3)H-serotonin or (3)H-choline. (35)S GTPgammaS binding, a measure of G protein activation, was studied in mouse and guinea pig hippocampal membranes. L-826,266 antagonised the effect of sulprostone on noradrenaline release in the rat cortex, yielding a Schild plot-based pA(2) value of 7.56. Apparent pA(2) values in mouse cortex and rat vas deferens (noradrenaline release) and rat cortex (serotonin release) were 7.55, 7.87 and 7.67, respectively. PGE(2) did not affect acetylcholine release in rat brain and dopamine release in guinea pig retina. In seven mice tissues, noradrenaline release was inhibited by sulprostone but not affected by virodhamine. (35)S GTPgammaS binding was not altered by sulprostone but stimulated by the cannabinoid agonist WIN 55,212-2. Prostaglandins of the E series inhibit monoamine release via EP(3) receptors at which L-826,266 is a competitive antagonist. EP(3) receptors that inhibit transmitter release are not present on central cholinergic neurones and retinal dopaminergic cells. Virodhamine is not converted to PGE(2). An EP(3) receptor model based on (35)S-GTPgammaS binding could not be identified. PMID- 20012266 TI - Metformin increases phagocytosis and acidifies lysosomal/endosomal compartments in AMPK-dependent manner in rat primary microglia. AB - Recent evidence suggests that metformin shows beneficial effects in experimental models of neuroinflammatory diseases. The aim of the present study was to determine the effect of metformin on phagocytosis and acidification of lysosomal/endosomal compartments in rat primary microglia in the presence of lipopolysaccharide (LPS) and/or beta-peptides (25-35), (1-40), and (1-42). Metformin increased the phagocytosis of fluorescent microspheres in the presence or absence of all the beta-peptides. However, the drug had no effect on the phagocytosis in LPS-stimulated microglia regardless of the presence of all the beta-peptides. Metformin acidified the lysosomal/endosomal compartments in the presence or absence of the beta-peptide 1-40 in both resting and activated microglia. To elucidate the mechanism of metformin action, we used 5 aminoimidazole-4-carboxamide-1-beta-D-ribofuranoside as an activator of adenosine monophosphate-activated protein kinase (AMPK) and compound C as a confirmed pharmacological inhibitor of AMPK. We have shown that metformin increased AMPK activity in microglial cells and that all observed effects are AMPK-dependent because the pretreatment of microglia with compound C reversed the effects of the drug. Since degradation of proteins in lysosomal/endosomal compartments depends largely on their phagocytosis and acidification, metformin may be beneficial in proteinopathies affecting the brain. PMID- 20012267 TI - Tramadol-induced seizurogenic effect: a possible role of opioid-dependent histamine H1 receptor activation-linked mechanism. AB - The present study has been designed to investigate the role of opioid receptors, mast cells, and histamine receptors (H(1) subtype) in the seizurogenic effect of tramadol on pentylenetetrazole-treated mice. A single injection of pentylenetetrazole (80 mg kg(-1)) was used to elicit seizure activity in mice. Seizures were assessed in terms of the time latency of the onset of Straub-like tail, onset of jerky movements of whole body, convulsions, and death. Tramadol administration (50 mg kg (-1)) caused a marked increase in seizurogenic activity of pentylenetetrazole as measured in terms of a significant decrease in the time latency of the onset of Straub-like tail, jerky movements of whole body, convulsions, and death. Moreover, prior administration of naloxone (2 mg kg(-1)), fexofenadine (100 mg kg(-1)), cetrizine, sodium cromoglycate, and ketotifen (10 mg kg(-1)), respectively, attenuated the seizurogenic activity that tramadol exerted on pentylenetetrazole-treated mice. Therefore, it may be suggested that tramadol exerts a seizurogenic effect on mice via an H(1) receptor activation linked pathway possibly through an opioid receptor-dependent release of histamine from the mast cells. PMID- 20012269 TI - Region-specific sex-dependent pattern of age-related changes of proximal femoral cancellous bone and its implications on differential bone fragility. AB - Despite evident interest in age-related bone changes, data on regional differences within the proximal femur are scarce. To date, there has been no comprehensive study on site-specific age-related changes in the trabecular architecture of three biomechanically important femoral subregions (medial neck, lateral neck, and intertrochanteric region) for both genders. In this study we investigated age-related deterioration in the trabecular architecture of those three subregions of the femoral neck for both genders. The research sample included 52 proximal femora (26 males, 26 females; age range, 26-96 years) from Forensic Department at University of Belgrade. Bone sections from the three regions of interest were scanned by micro-CT at University of Hamburg. The study revealed that proximal femoral microarchitecture cannot be perceived as homogeneous and, more importantly, that the aging process is not uniform. Besides the initial intersite differences, microarchitecture changed differently with increasing age, maintaining significant differences between the regions. In addition, we observed a different aging pattern between genders: deterioration was most significant in the intertrochanteric region in women, while the lateral neck was most affected in men. This finding supports epidemiological data about the differential occurrence of cervical vs. trochanteric fractures in aging males and females. In conclusion, the aging process in the proximal femur cannot be regarded as a simple function of quantitative bone loss but, rather, as an alteration of specific architecture that may degrade bone strength. PMID- 20012268 TI - Reversal of propoxur-induced impairment of memory and oxidative stress by 4' chlorodiazepam in rats. AB - Carbamate pesticides like propoxur have been shown to adversely affect memory and induce oxidative stress on both acute and chronic exposure. The present study was designed to explore the modulation of the effects of propoxur over cognitive function by progesterone (PROG) and 4'-chlorodiazepam (4CD). Cognitive function was assessed using step-down latency (SDL) on a passive avoidance apparatus, transfer latency (TL) on a plus maze and spatial navigation test on Morris water maze. Oxidative stress was assessed by examining brain malondialdehyde (MDA) and reduced glutathione (GSH) levels and catalase (CAT) activity. A significant reduction in SDL and prolongation of TL and spatial navigation test was found for the propoxur (10 mg/kg/d; p.o.) treated group at weeks 6 and 7 as compared with control. One-week treatment with 4CD (0.5 mg/kg/d; i.p.) antagonized the effect of propoxur on SDL, spatial navigation test as well as TL; whereas, PROG failed to modulate this effect at a dose of 15 mg/kg/d, i.p. Propoxur produced a statistically significant increase in the brain MDA levels and decrease in the brain GSH levels and CAT activity. Treatment with 4CD at the above dose attenuated the effect of propoxur on oxidative stress whereas PROG (15 mg/kg/d; i.p.) failed to influence the same. The results of the present study thus show that 4-CD has the potential to attenuate cognitive dysfunction and oxidative stress induced by toxicants like propoxur in the brain. PMID- 20012270 TI - Evidence of reduced bone turnover and disturbed mineralization process in a boy with Stickler syndrome. AB - We describe a tall-statured 14-year-old boy who illustrated the full phenotypic and radiographic features of Stickler syndrome type I. A bone biopsy showed evidence of reduced bone mass and bone turnover, such as reduced BV/TV (-43%), TbTh (-29%), and OS/BS (-48%), Ob.S/BS (-27%), and Oc/BS (-47%) compared to "age matched" controls. Moreover, there was evidence that the mineralization process was severely disturbed. Quantitative backscattered electron imaging revealed that the bone mineralization density distribution (BMDD) of cancellous (Cn) as well as cortical (Ct) bone was shifted toward lower mineralization compared to a young control reference cohort. BMDD parameters of mean degree of mineralization, Cn Ca (-9.8%) and Ct Ca (-18.0%), were dramatically decreased. To the best of our knowledge this is the first clinical report describing bone biopsy findings in a boy with Stickler syndrome. Such a severe undermineralization of bone matrix might essentially contribute to the compromised mechanical competence of the skeleton found in this patient. PMID- 20012271 TI - Evolutionary analysis of mammalian enamelin, the largest enamel protein, supports a crucial role for the 32-kDa peptide and reveals selective adaptation in rodents and primates. AB - Enamelin (ENAM) plays an important role in the mineralization of the forming enamel matrix. We have performed an evolutionary analysis of mammalian ENAM to identify highly conserved residues or regions that could have important function (selective pressure), to predict mutations that could be associated with amelogenesis imperfecta in humans, and to identify possible adaptive evolution of ENAM during 200 million years ago of mammalian evolution. In order to fulfil these objectives, we obtained 36-ENAM sequences that are representative of the mammalian lineages. Our results show a remarkably high conservation pattern in the region of the 32-kDa fragment of ENAM, especially its phosphorylation, glycosylation, and proteolytic sites. In primates and rodents we also identified several sites under positive selection, which could indicate recent evolutionary changes in ENAM function. Furthermore, the analysis of the unusual signal peptide provided new insights on the possible regulation of ENAM secretion, a hypothesis that should be tested in the near future. Taken together, these findings improve our understanding of ENAM evolution and provide new information that would be useful for further investigation of ENAM function as well as for the validation of mutations leading to amelogenesis imperfecta. PMID- 20012273 TI - Evaluation and validation of two chromatographic methods (HPLC-fluorescence and LC-MS/MS) for the determination and confirmation of ochratoxin A in pig tissues. AB - Two different analytical methods for the determination and confirmation of ochratoxin A (OTA) in blood serum, kidney, and liver of pigs have been compared. Sample cleanup was based on liquid-liquid phase extraction. The detection of OTA was accomplished with high-performance liquid chromatography (HPLC) combined either with fluorescence detection (FLD) or electrospray ionization (ESI+) tandem mass spectrometry (MS/MS). The comparative method of evaluation was based on the investigation of 90 samples of blood serum, kidney, and liver per animal originating from different regions of Serbia. The analytical results are discussed in view of the respective method validation data and the corresponding experimental protocols. In general, analytical data obtained with liquid chromatography (LC)-MS/MS detection offered comparable good results in the sub ppb concentration level indicating that the electrospray tandem mass spectrometric (LC-MS/MS) method was more selective and sensitive for the analysis and confirmation of OTA in pig tissues than the HPLC method after the methylation of OTA. PMID- 20012272 TI - The NfeD protein family and its conserved gene neighbours throughout prokaryotes: functional implications for stomatin-like proteins. AB - NfeD-like proteins are widely distributed throughout prokaryotes and are frequently associated with genes encoding stomatin-like proteins (slipins). Here, we reveal that the NfeD family is ancient and comprises three major groups: NfeD1a, NfeD1b and truncated NfeD1b. Members of each group are associated with one of four conserved gene partners, three of which have eukaryotic homologues that are membrane raft associated, namely stomatin, paraslipin (previously SLP-2) and flotillin. The first NfeD group (NfeD1b), comprises proteins of approximately 460-aa long that have three functional domains: an N-terminal protease, a middle membrane-spanning region and a soluble C-terminal region rich in beta-strands. The nfeD1b gene is adjacent to eoslipin in prokaryotic genomes except in Firmicutes and Deinococci, where yqfA replaces eoslipin. Proteins in the second major group (NfeD1a) are homologous to the C-terminus of NfeD1b which forms a beta-barrel-like domain, and their genes are associated with paraslipin. Using OrthoMCL clustering, we show that nfeD1b genes have become truncated on many independent occasions giving rise to the third major group. These short NfeD homologues frequently remain associated with their ancestral gene neighbour, resembling NfeD1a in structure, yet are much more related to full-length NfeD1b; we term these "truncated NfeD1b". These conserved associations suggest that NfeD proteins are dependent on gene partners for their function and that the site of interaction may lie within the C-terminal portion that is common to all NfeD homologues. Although NfeD homologues are confined to prokaryotes, this conserved association could represent an excellent system to study slipin and flotillin proteins. PMID- 20012274 TI - Genetic, biochemical, and individual responses of the teleost fish Carassius auratus to uranium. AB - Carassius auratus were exposed for 96 h to different concentrations of uranyl nitrate (corresponding to 0, 100, 450, and 2,025 microg U L(-1)) and killed after different postexposure periods (0, 48, and 96 h) to assess uranium bioaccumulation, peroxisome proliferation (catalase [CAT]), lipid peroxidation (thiobarbituric acid reactive substances [TBARS]), and DNA integrity in erythrocytes (comet assay). In addition, feeding behaviour was recorded as a general response to toxicant exposure. Results provided evidence of uranium bioaccumulation in muscle of C. auratus after exposure to the highest concentrations (450 and 2,025 microg U L(-1)). This tissue was able to depurate uranium to control levels 96 h after exposure ceased. However, no perturbations in feeding behaviour or cell damage were observed in the tested organisms, except for the apparent irreversible inhibition of CAT activity immediately after exposure in the highest concentration tested. Data on DNA integrity (comets) showed that waterborne uranium exposure was able to induce genotoxicity in C. auratus erythrocytes because fish exposed to all concentrations exhibited higher DNA damage than controls 96 h after exposure. No DNA damage repair was apparent throughout the postexposure period, which was contrary to a recovery scenario. This experiment provides evidence of uranium's ability to induce physiologic impairment and genotoxicity in freshwater fish at environmentally relevant concentrations. PMID- 20012276 TI - Current state of coenzyme Q(10) production and its applications. AB - Coenzyme Q(10) (CoQ(10)), an obligatory cofactor in the aerobic respiratory electron transfer for energy generation, is formed from the conjugation of a benzoquinone ring with a hydrophobic isoprenoid chain. CoQ(10) is now used as a nutritional supplement because of its antioxidant properties and is beneficial in the treatment of several human diseases when administered orally. Bioprocesses have been developed for the commercial production of CoQ(10) because of its increased demand, and these bioprocesses depend on microbes that produce high levels of CoQ(10) naturally. However, as knowledge of the biosynthetic enzymes and the regulatory mechanisms modulating CoQ(10) production increases, approaches arise for the genetic engineering of CoQ(10) production in Escherichia coli and Agrobacterium tumefaciens. This review focused on approaches for CoQ(10) production, strategies used to engineer CoQ(10) production in microbes, and potential applications of CoQ(10). PMID- 20012275 TI - A large-scale epidemiological study to identify bacteria pathogenic to Pacific oyster Crassostrea gigas and correlation between virulence and metalloprotease like activity. AB - A 4-year bacteriological survey (2003-2007) of four molluscs cultivated in France and faced with mortality episodes was performed by the French shellfish pathology network. The more abundant bacteria isolated during 92 mortality episodes, occurring mainly in Pacific oyster Crassostrea gigas, were identified by genotyping methods. It allowed us both to confirm the representativeness of Vibrio splendidus and Vibrio aestuarianus bacterial strains and to identify both a large number of Vibrio harveyi-related strains mainly detected during 2007 oyster mortality outbreaks and to a lesser extent bacterial strains identified as Shewanella colwelliana. Because metalloprotease has been reported to constitute a virulence factor in a few Vibrio strains pathogenic for C. gigas, several bacterial strains isolated in this study were screened to evaluate their pathogenicity in C. gigas spat by experimental infection and their ability to produce metalloprotease-like activity in the culture supernatant fluids. A high level (84%) of concordant results between azocaseinase activities and virulence of strains was obtained in this study. Because bacterial metalloprotease activities appeared as a common feature of pathogenic bacteria strains associated with mortality events of C. gigas reared in France, this phenotypic test could be useful for the evaluation of virulence in bacterial strains associated with such mortality episodes. PMID- 20012277 TI - Production and characterization of a recombinant single-chain antibody against Hantaan virus envelop glycoprotein. AB - Hantaan virus (HTNV) is the type of Hantavirus causing hemorrhagic fever with renal syndrome, for which no specific therapeutics are available so far. Cell type-specific internalizing antibodies can be used to deliver therapeutics intracellularly to target cell and thus, have potential application in anti-HTNV infection. To achieve intracellular delivery of therapeutics, it is necessary to obtain antibodies that demonstrate sufficient cell type-specific binding, internalizing, and desired cellular trafficking. Here, we describe the prokaryotic expression, affinity purification, and functional testing of a single chain Fv antibody fragment (scFv) against HTNV envelop glycoprotein (GP), an HTNV specific antigen normally located on the membranes of HTNV-infected cells. This HTNV GP-targeting antibody, scFv3G1, was produced in the cytoplasm of Escherichia coli cells as a soluble protein and was purified by immobilized metal affinity chromatography. The purified scFv possessed a high specific antigen-binding activity to HTNV GP and HTNV-infected Vero E6 cells and could be internalized into HTNV-infected cells probably through the clathrin-dependent endocytosis pathways similar to that observed with transferrin. Our results showed that the E. coli-produced scFv had potential applications in targeted and intracellular delivery of therapeutics against HTNV infections. PMID- 20012278 TI - Reconstruction of the violacein biosynthetic pathway from Duganella sp. B2 in different heterologous hosts. AB - Violacein is a bacteria-originated indolocarbazole pigment with potential applications due to its various bioactivities such as anti-tumor, antiviral, and antifungal activities. However, stable mass production of this pigment is difficult due to its low productivities and the instability of wild-type violacein-producing strains. In order to establish a stable and efficient production system for violacein, the violacein synthesis pathway from a new species of Duganella sp. B2 was reconstructed in different bacterial strains including Escherichia coli, Citrobacter freundii, and Enterobacter aerogenes by using different vectors. The gene cluster that encodes five enzymes involved in the violacein biosynthetic pathway was first isolated from Duganella sp. B2, and three recombinant expression vectors were constructed using the T7 promoter or the alkane-responsive promoter PalkB. Our results showed that violacein could be stably synthesized in E. coli, C. freundii, and E. aerogenes. Interestingly, we found that there were great differences between the different recombinant strains, not only in the protein expression profiles pertaining to violacein biosynthesis but also in the productivity and composition of crude violacein. Among the host strains tested, the crude violacein production by the recombinant C. freundii strain reached 1.68 g L(-1) in shake flask cultures, which was 4-fold higher than the highest production previously reported in flask culture by other groups. To the best of our knowledge, this is the first report on the efficient production of violacein by genetically engineered strains. PMID- 20012279 TI - Enzymatic and whole-cell synthesis of lactate-containing polyesters: toward the complete biological production of polylactate. AB - The importance of polylactic acid, a representative bio-based polyester, has been established on a worldwide scale in response to emerging global environmental problems such as green house gas emission and limited petroleum consumption. The current methods for generating this bio-based polymer involve biological synthesis and lactic acid (LA) fermentation, followed by chemical ring-opening polymerization. Among the research community working on polyhydroxyalkanoate polyesters, the prospect of direct biological synthesis of LA into a polymeric form is very attractive from the academic and industrial perspectives. In 2008, this challenge was met for the first time by the discovery of an "LA-polymerizing enzyme". Using this novel enzyme, the metabolic engineering approach outlined here provided an entirely new, single organism generation of the polymer. This is a major breakthrough in the field. In this review, we provide an overview of the whole-cell synthesis of LA-containing polyesters in comparison with conventional lipase-catalyzed polymer synthesis in terms of both the concepts and strategies of their synthetic processes. PMID- 20012280 TI - Xylitol production by recombinant Corynebacterium glutamicum under oxygen deprivation. AB - Wild-type Corynebacterium glutamicum produced 0.6 g l(-1) xylitol from xylose at a productivity of 0.01 g l(-1) h(-1) under oxygen deprivation. To increase this productivity, the pentose transporter gene (araE) from C. glutamicum ATCC31831 was integrated into the C. glutamicum R chromosome. Consequent disruption of its lactate dehydrogenase gene (ldhA), and expression of single-site mutant xylose reductase from Candida tenuis (CtXR (K274R)) resulted in recombinant C. glutamicum strain CtXR4 that produced 26.5 g l(-1) xylitol at 3.1 g l(-1) h(-1). To eliminate possible formation of toxic intracellular xylitol phosphate, genes encoding xylulokinase (XylB) and phosphoenolpyruvate-dependent fructose phosphotransferase (PTS(fru)) were disrupted to yield strain CtXR7. The productivity of strain CtXR7 increased 1.6-fold over that of strain CtXR4. A fed batch 21-h CtXR7 culture in mineral salts medium under oxygen deprivation yielded 166 g l(-1) xylitol at 7.9 g l(-1) h(-1), representing the highest bacterial xylitol productivity reported to date. PMID- 20012281 TI - Regulation of genes in Streptomyces bacteria required for catabolism of lignin derived aromatic compounds. AB - The major utilization pathway for lignin-derived aromatic compounds in microorganisms is the beta-ketoadipate pathway. Through this pathway, the aromatic compounds protocatechuate and catechol are converted to acetyl coenzyme A and succinyl coenzyme A. The enzymes of the protocatechuate branch of this pathway are encoded by the pca genes. Here, we describe a gene cluster in Streptomyces coelicolor containing the pca structural genes and a regulatory gene required for the catabolism of protocatechuate. We found that transcription of the structural genes in S. coelicolor is induced by protocatechuate and p hydroxybenzoate. We also observed inducible transcription of pca structural genes in the ligninolytic strain Streptomyces viridosporus ATCC 39115. Disruption of a gene encoding a putative MarR family transcription factor that is divergently transcribed from the pca structural genes resulted in constitutive transcription of the structural genes. Thus, the transcription factor encoded by this gene is an apparent negative regulator of pca gene transcription in S. coelicolor. Our findings suggest how Streptomyces bacteria could be engineered for and used in biotechnology for the utilization and degradation of lignin and lignin-derived aromatic compounds. PMID- 20012282 TI - Secretion of Kluyveromyces lactis Cu/Zn SOD: strategies for enhanced production. AB - The Kluyveromyces lactis Cu/Zn SOD gene (SOD1) was fused with the toxin K1 signal sequence to obtain extracellular production of superoxide dismutase. Kluyveromyces marxianus L3 and K. lactis MW98-8C strains were transformed and compared as hosts for the secretion. The effects of the media composition were evaluated: In K. lactis, the highest volumetric activity was obtained in YKK synthetic medium in the presence of Cu(2+)/Zn(2+) cofactors (9.6 kU l(-1)). In K. marxianus, active SOD was produced only in YPD medium supplemented with Cu(2+) and Zn(2+) (8.8 kU l(-1)). In order to improve the production of secreted active SOD in K. lactis, the SOD1 copper carrier (CCS1) was overexpressed and targeted to the secretory apparatus. A positive effect was observed only when K. lactis was grown in a medium without Cu(2+)/Zn(2+) supplement. The best performing culture conditions for K. lactis and K. marxianus recombinant strains were successfully applied to two laboratory-scale fed-batch processes, and volumetric SOD activities increased up to 19.4 and 24.1 kU l(-1), respectively. PMID- 20012283 TI - A clinical perspective on the criteria for liver resection and the use of liver function tests. PMID- 20012284 TI - Hyperbaric oxygen therapy as an adjunct to surgical treatment of extensive hidradenitis suppurativa. PMID- 20012285 TI - Relationship between the histological type of initial lesions and the risk for the development of remnant gastric cancers after gastrectomy for synchronous multiple gastric cancers. AB - BACKGROUND: Clinicopathological features of remnant gastric cancers after gastrectomy for synchronous multiple gastric cancers have not been fully addressed. In this study we evaluated the potential risk factors attributable to the development of remnant gastric cancers after initial gastrectomy for multiple gastric cancers. METHODS: We retrospectively analyzed 3042 patients diagnosed with gastric cancers who underwent a gastrectomy. Of these, 922 total gastrectomy cases were excluded and the remaining 2120 cases were clinicopathologically investigated. RESULTS: Among the 2120 patients, 1967 patients were histopathologically diagnosed with a solitary lesion and 153 patients with multiple lesions. The incidence of remnant gastric cancers was higher in patients with multiple lesions at initial surgery than in those with a solitary lesion (7/153 [4.5%] vs. 9/1967 [0.45%], p < 0.05). Among the patients with remnant gastric cancer, the percentages of cases with a combination of undifferentiated type carcinoma (Type C), differentiated-type carcinoma (Type A), and differentiated- and undifferentiated-type carcinoma (Type B) as initial lesions were 15.0, 3.3, and 2.7%, respectively. Compared with those with no remnant gastric cancer, the incidence of the combination of undifferentiated-type carcinoma (Type C) as an initial lesion was significantly associated with a higher (p < 0.05) incidence of remnant gastric cancer. CONCLUSION: As initial lesions, a combination of undifferentiated-type carcinoma was a potential risk factor for the development of remnant gastric cancers after initial gastrectomy. PMID- 20012286 TI - Laparoscopic liver resection for hepatocellular carcinoma in the left liver: Pringle maneuver versus tourniquet method. AB - BACKGROUND: A good postoperative outcome after partial hepatectomy is highly dependent on limiting operative blood loss. This study evaluated the feasibility and efficacy of the tourniquet method compared with the Pringle maneuver in laparoscopic liver resection for hepatocellular carcinoma (HCC) in the left liver. METHODS: A retrospective, nonrandomized, comparative study for laparoscopic liver resection for HCC in the left liver using the Pringle maneuver (group A) or the tourniquet method (group B) was initiated in our center between March 2004 and October 2008. RESULTS: Sixteen patients (group A) underwent laparoscopic liver resection using the Pringle maneuver, and 13 patients (group B) underwent laparoscopic liver resection using the tourniquet method. No differences in operation time, operative blood loss, perioperative blood transfusion, and perioperative morbidity were found between the two groups. Both groups had no postoperative mortality. The liver enzymes were significantly elevated in group A compared with group B. Group B patients also had significantly faster recovery of liver function. The postoperative hospital stay for group B was significantly shorter than group A (mean, 5.6 days vs. 8.3 days). CONCLUSIONS: Both techniques of vascular control were equally safe, efficacious, and feasible for patients undergoing laparoscopic left-sided liver resection. The tourniquet method gave a wider safety margin for patients with chronic liver disease with a compromised hepatic reserve by causing less ischemia-reperfusion injury to the remnant liver. PMID- 20012287 TI - Mortality-related factors in 1056 radioiodine-treated patients with well differentiated thyroid cancer in southern Thailand. AB - BACKGROUND: Reported risk factors for death related to well differentiated thyroid cancer vary, possibly depending on the methods and geographical site of data collection. METHOD: To evaluate variables related to death in well differentiated thyroid cancer (WDTC) in a population of southern Thailand, we retrospectively reviewed medical records of all those treated with radioactive iodine (I-131) at the Division of Nuclear Medicine from 1983 to 2002. We found 1056 subjects. Variables examined included present status of subjects (transfer, lost, died), cause of death (related and not related to WDTC), duration of follow up since surgery, largest size of tumor (when available), gender, histology (follicular or papillary, with mixed included in papillary), age of surgery, and site of spread when treated. Survival from WDTC death utilized Kaplan-Meier and Cox regression analysis. RESULTS: The median follow-up was 7 years, while only 4 years elapsed from surgery for those who died. We did not know the status of 16% of the subjects despite attempts to contact them. Survival was not affected by age, gender, or maximum size of tumor. Age at surgery was significant only for the group older than 55 years, with a 10-year survival of 76%. The spread of the disease that affected survival can be localized to the aerodigestive tract (10 year survival of 65%) and metastases associated with bone fracture (10-year survival 34%), while other metastases (including asymptomatic bone metastases) exhibited a 10-year survival of 93%. Neither lymph node spread nor elevated Tg at maximum stimulation predicted survival. CONCLUSIONS: To better predict survival of WDTC, site of spread and degree of spread has to be specific. PMID- 20012288 TI - Development and evaluation of an observational tool for assessing surgical flow disruptions and their impact on surgical performance. AB - BACKGROUND: Many researchers have previously explored the correlation between surgical flow disruptions and adverse events in cardiac surgery; however, there is no reliable tool to prospectively categorize surgical flow disruptions and the conditions that predispose a surgical team to adverse events. METHODS: Two independent raters of different medical and human factors expertise observed 12 cardiovascular operations and iteratively designed a surgical flow disruption tool (SFDT) to characterize surgical flow disruptions and the latent factors that contribute to adverse events. Categories to characterize surgical flow disruptions were created based on human factors models of human error. After the design period, both raters observed ten surgical cases using the tool to assess validity and inter-rater reliability. RESULTS: Rating agreement (weighted kappa) for each category across the ten surgeries was moderate to very high, resulting in strong inter-rater reliability for each category on the surgical flow disruption tool. Use of the SFDT was simple and clear for observers of diverse backgrounds, including human factors experts and medical personnel. CONCLUSIONS: This research depicts the development and utility of a tool to analyze surgical flow disruptions in the cardiovascular operating room with satisfactory inter rater reliability. This tool is an important first step in systematically categorizing and measuring surgical flow disruptions and their impact on patient safety in the operating room. PMID- 20012290 TI - Risks and adequacy of an optimized surgical approach to the primary surgical management of papillary thyroid carcinoma treated during 1999-2006. AB - BACKGROUND: Intense disease surveillance and frequent lymph node metastases (LNMs) in papillary thyroid cancer (PTC) have resulted in increased locoregional recurrences. We examined the safety and efficacy of an optimized surgical approach including preoperative ultrasonography (US), bilateral thyroidectomy, routine compartment VI dissection, and lateral neck dissection for LNM. METHODS: During 1999-2006, a total of 420 patients underwent optimized primary surgery; 291(69%) females, median age 46 years; follow-up 98%, median 4.4 years. Patients were reviewed for tumor characteristics, pattern of LNM, staging, and outcomes. RESULTS: Total or near-total thyroidectomy was performed in 212 (51%) and 208 (49%) patients, respectively. Tumors were multicentric, 40% (average 1.7 cm); were bilateral, 30%; and showed extrathyroidal extension, 17%. Overall, 223 (53%) patients had LNMs: 213 (51%) were central and 85 (20%) were lateral jugular. pTNM staging: I, 258 (61%); II, 35 (8%); III, 88 (21%); IV, 39 (9%). AGES (age, grade, extension, and size-thyroid tumor; and MACIS (metastasis, age, completeness of resection, invasion, and size) prognostic scores were low risk in 362 (86%) and 352 (84%), respectively. Relapse developed in 57 (14%) patients: LNM in 44, soft tissue local recurrence (LR) in 5, distant metastases (DM) in 8. Hypoparathyroidism occurred in 5 (1.2%) patients and 1 had unintentional laryngeal nerve damage. Relapse with LNM occurred in previously operated fields in 19 (5%) patients, 11(3%) from disease virulence (LR or DM), preoperative false negative (FN) US in 12 (3%), and combination of FN-US and recurrence in the operated field in 5 (1%) patients. CONCLUSIONS: Recurrence was limited to 5% of patients when the extent of disease was accurately defined and potentially curable. This optimized surgical strategy is relatively safe. PMID- 20012291 TI - Comparison of consumer information on the internet to the current evidence base for minimally invasive parathyroidectomy. AB - BACKGROUND: The Internet is increasingly used as a source of health information by patients. Under these circumstances, the opportunity exists for Internet sites ostensibly providing patient information to act to promote surgical referrals based on exaggerated claims. This study aims to assess quantitatively and qualitatively the Internet-based consumer health information for minimally invasive parathyroidectomy (MIP) techniques. METHODS: This is a prospective analysis of Internet web sites. Descriptive information about specific published claims on each of the web sites was documented and compared to the published evidence base. Web sites were then rated using a validated composite score (CS) tool and an MIP score tool developed specifically for the study. RESULTS: The search yielded 308 web sites, which, after assessment by the inclusion criteria left 44 unique web sites suitable for analysis. "Exaggerated," "misleading," or "false" claims were present in 27.3% of the web sites analyzed. The false claims category had a high negative item-total correlation with the overall score, and accuracy was found to have a statistically significant (p < 0.05) negative correlation with quality. However, analysis performed for country of origin and the organization responsible for the web site found no significant difference. CONCLUSIONS: Web sites offering information in relation to MIP have a surprisingly high rate of claims that are not in accord with the evidence. Such claims may be posted to attract surgical referrals. It is difficult for consumers to differentiate quality consumer health web sites from poor ones as there are no hard and fast rules to differentiate them. PMID- 20012292 TI - Synergistic interactions between aminoflavone, paclitaxel and camptothecin in human breast cancer cells. AB - PURPOSE: Aminoflavone is a unique DNA damaging agent currently undergoing phase I evaluation in a prodrug form (AFP464). In anticipation of combination regimens, interactions between aminoflavone and several anticancer drugs were investigated in MCF-7 breast cancer cells to determine whether synergistic cancer cell killing effects were observed. METHODS: Colony formation assays were performed to assess the effect of combining aminoflavone with a variety of anticancer drugs. Changes in initial uptake, retention or efflux of aminoflavone and the second agent were compared to the behavior of drugs alone. Key features required for aminoflavone activity in cell culture models were also explored, focusing on the obligatory induction of CYP1A1/1A2 and binding of reactive aminoflavone metabolites to tumor cell total macromolecules and DNA. RESULTS: Aminoflavone was synergistic when co incubated with paclitaxel, camptothecin or SN38. Uptake of neither aminoflavone nor any of the other three compounds was altered in combination incubations. Paclitaxel did not inhibit DNA binding of aminoflavone metabolites, while camptothecin did. Aminoflavone-induced CYP1A1 induction was observed in the presence of camptothecin or paclitaxel. CONCLUSIONS: Aminoflavone is a promising therapeutic agent for breast cancer due to its unique mechanism of action compared to commonly used drugs. Combined treatments utilizing aminoflavone in conjunction with paclitaxel or camptothecin may provide an even greater cytotoxic effect than achieved with aminoflavone alone. PMID- 20012293 TI - A phase I trial of PR-104, a nitrogen mustard prodrug activated by both hypoxia and aldo-keto reductase 1C3, in patients with solid tumors. AB - PURPOSE: PR-104 is a "pre-prodrug" designed to be activated to a dinitrobenzamide nitrogen mustard cytotoxin by nitroreduction in hypoxic regions of tumors. This study was conducted to establish the maximum tolerated dose (MTD), dose-limiting toxicity (DLT), safety, and pharmacokinetics (PK) of PR-104 in patients with advanced solid tumors. METHODS: Patients with solid tumors refractory or not amenable to conventional treatment were evaluated in a dose-escalation trial of PR-104 administered as a 1-h intravenous (IV) infusion every 3 weeks. The plasma PK of PR-104 and its primary metabolite, PR-104A, were evaluated. RESULTS: Twenty seven patients received a median of two cycles of PR-104 in doses ranging from 135 to 1,400 mg/m(2). The MTD of PR-104 as a single-dose infusion every 3 weeks was established as 1,100 mg/m(2). One of six patients treated at 1,100 mg/m(2) experienced DLT of grade 3 fatigue. Above the MTD, the DLTs at 1,400 mg/m(2) were febrile neutropenia and infection with normal absolute neutrophil count. No objective responses were observed, although reductions in tumor size were observed in patients treated at doses > or = 550 mg/m(2). The plasma PK of PR-104 demonstrated rapid conversion to PR-104A, with approximately dose-linear PK of both species. CONCLUSIONS: PR-104 was well tolerated at a dose of 1,100 mg/m(2) administered as an IV infusion every 3 weeks. The area under the PR-104A plasma concentration-time curve at this dose exceeded that required for activity in human tumor cell cultures and xenograft models. The recommended dose of PR-104 as a single agent for phase II trials is 1,100 mg/m(2) and further trials are underway. PMID- 20012294 TI - RAD59 and RAD1 cooperate in translocation formation by single-strand annealing in Saccharomyces cerevisiae. AB - Studies in the budding yeast, Saccharomyces cerevisiae, have demonstrated that a substantial fraction of double-strand break repair following acute radiation exposure involves homologous recombination between repetitive genomic elements. We have previously described an assay in S. cerevisiae that allows us to model how repair of multiple breaks leads to the formation of chromosomal translocations by single-strand annealing (SSA) and found that Rad59, a paralog of the single-stranded DNA annealing protein Rad52, is critically important in this process. We have constructed several rad59 missense alleles to study its function more closely. Characterization of these mutants revealed proportional defects in both translocation formation and spontaneous direct-repeat recombination, which is also thought to occur by SSA. Combining the rad59 missense alleles with a null allele of RAD1, which encodes a subunit of a nuclease required for the removal of non-homologous tails from annealed intermediates, substantially suppressed the low frequency of translocations observed in rad1-null single mutants. These data suggest that at least one role of Rad59 in translocation formation by SSA is supporting the machinery required for cleavage of non-homologous tails. PMID- 20012295 TI - Regional lymph node metastasis and locoregional recurrence of rectal carcinoma in the era of TME [corrected] surgery. Implications for treatment decisions. AB - BACKGROUND AND AIMS: For rectal carcinoma treated according to the concept of total mesorectal excision (TME surgery), the independent influence of regional lymph node metastasis on the locoregional recurrence risk is still in discussion. A reliable assessment of this risk is important for an individualised selective indication for neoadjuvant radio-/radiochemotherapy. METHODS: Analysis of literature, especially of the last 20 years, and consideration of pathological and oncological basic research. Multivariate analysis of data of the Erlangen Registry of Colorectal Carcinoma. RESULTS: The clinical assessment of the pretherapeutic regional lymph node status by the present available imaging methods is still unreliable. The analysis of the association between pretherapeutic regional lymph node status and locoregional recurrence risk has to be based on follow-up data of patients treated by primary surgery and has to be distinguished between patients treated by conventional and optimised quality assured TME surgery, respectively. Data from Erlangen show an increase of the local recurrence risk for patients with at least four involved regional lymph nodes. CONCLUSIONS: For patients with at least four involved regional lymph nodes, a neoadjuvant radiochemotherapy may be indicated. However, today, the pretherapeutic diagnosis is uncertain and results in overtherapy in 40%. Thus, in case of positive lymph node findings by imaging methods, the benefits and risk of neoadjuvant therapy in such situations should always be discussed with the patient in the sense of informed consent and shared decision. PMID- 20012297 TI - Irinotecan and colorectal cancer: the role of p53, VEGF-C and alpha-B-crystallin expression. PMID- 20012296 TI - Mechanical bowel preparation does not affect the intramucosal bacterial colony count. AB - PURPOSE: The aim of this study was to determine if mechanical bowel preparation (MBP) influences the intramucosal bacterial colony count in the colon. MATERIALS AND METHODS: Macroscopically normal colon mucosa was collected from 37 patients (20 with and 17 without MBP) who were undergoing elective colorectal surgery at three hospitals. The biopsies were processed and cultured in the same laboratory. Colony counts of the common pathogens Escherichia coli and Bacteroides as well as of total bacteria were conducted. The study groups were comparable with regard to age, gender, antibiotics use, diagnosis and type of resection. RESULTS: MBP did not influence the median colony count of E. coli, Bacteroides or total bacteria in our study. CONCLUSIONS: MBP did not affect the intramucosal bacterial count in this study. Further studies are suggested to confirm these findings. PMID- 20012298 TI - Colovesical, coloenteric, colocutaneous fistula, and hip septic arthritis secondary to sigmoid diverticulitis. PMID- 20012299 TI - Chylous ascites secondary to pseudomembranous colitis following ileostomy reversal fashioned for low anterior resection. PMID- 20012300 TI - Carbachol induces TGF-alpha expression and colonic epithelial cell proliferation in sensory-desensitised rats. AB - OBJECTIVE/BACKGROUND: Signals for the expression of the peptide growth factors epidermal growth factor and transforming growth factor-alpha (TGFalpha) in the gastrointestinal mucosa are largely unknown. We have shown earlier that extrinsic afferents in the gastrointestinal tract induce TGFalpha expression in colonic mucosa via the deliberation of neurotransmitters substance P and calcitonin gene related peptide. The aim of our present study was to determine the effects of carbachol on mucosal TGFalpha expression and epithelial cell proliferation in vivo. DESIGN/METHODS: Rats were divided in three groups. Group 1 was treated with vehicle only, group 2 received one single subcutaneous injection of 250 microg/kg of carbachol and animals in group 3 were sensory-desensitised prior to the injection of 250 microg/kg carbachol. TGFalpha expression and epithelial cell proliferation was evaluated by polymerase chain reaction, Western blot analysis and bromodeoxyuridine staining. RESULTS: Carbachol induced a significant increase in mucosal epithelial cell proliferation and TGFalpha expression. Sensory desensitisation did neither abolish the increased TGFalpha expression nor the increase in epithelial cell proliferation. CONCLUSION: Parasympathetic pathways are involved in the control of TGFalpha expression in gastrointestinal mucosa as well as in epithelial cell proliferation. PMID- 20012302 TI - [Oral hygiene, prophylaxis and therapy in patients with inflammatory rheumatic diseases]. AB - Inflammatory disorders of the periodontium are often associated with chronic systemic diseases, which can demonstrate a reciprocal influence. Within the adult population at present, 74% of younger adults and 88% of older individuals require periodontal treatment. Due to inflammatory processes, patients with rheumatoid arthritis or other chronic polyarthritides frequently suffer from pain in the temporomandibular joint and, since finger mobility is often limited, their ability to perform oral hygiene measures is impeded. However, diligent and constant oral hygiene is of crucial importance both for maintaining a healthy periodontium and to prevent the development of caries. For their daily dental care, these patients should favor the use of electric toothbrushes, products for interdental cleaning and mouth rinses. The dentist should be informed immediately about increased and constant gingival bleeding, gingival hyperplasia, loosening or migration of teeth associated with gingival recession or other irritations in the oral cavity. Professional dental cleaning should be routinely performed at 3- to 6-month intervals in order to prevent an increase in the risk for periodontal disease. PMID- 20012301 TI - Functional analysis of LOXL2 in pancreatic carcinoma. AB - OBJECTIVES: Lysyl oxidase-like 2 (LOXL2) plays a part in epithelial-mesenchymal transition (EMT) by stabilizing the transcription factor SNAI1. Previous studies showed that LOXL2 is one of the most highly and specifically upregulated genes in pancreatic cancer. LOXL2 was also found to be strongly upregulated in the secretome of established pancreatic cancer cell lines. To get more insight into the aggressive growth and infiltrating nature of pancreatic cancer, we evaluated the functional role of LOXL2 in pancreatic cancer cells. METHODS: Gene inhibition by small interfering RNAs was used to silence LOXL2 in pancreatic cancer cell lines MiaPaCa-2 and Panc1. Cell death, proliferation, and morphology of transfected cells were determined. Cell characteristics under cell stress and gemcitabine treatment were analyzed. Gene expression analysis of transfected cells by DNA microarray was used to understand the processes of chemosensitization. RESULTS: Silencing of LOXL2 in pancreatic cancer cells resulted in an augmented sensitivity toward gemcitabine treatment, with significantly elevated cell death and reduced viable cells. However, transfection had no direct effect on morphology or growth pattern of Mia PaCa-2 and Panc1 cell lines. Gene expression analysis identified among others the transcription factor E2F5 as possible target of LOXL2. CONCLUSIONS: Gene inhibition of the EMT regulatory gene LOXL2 resulted in a distinct sensitization toward gemcitabine. Additionally, gene expression analysis showed a role for LOXL2 in the regulation of different transcription factors associated with invasion and metastasis. Our results suggest that the improved response toward chemotherapy in LOLX2-silenced pancreatic cancer cells is possibly mediated by the transcription factor E2F5. PMID- 20012303 TI - [Effect of increasing the infliximab dose in rheumatoid arthritis]. PMID- 20012304 TI - Pomeroy technique or Filshie clips for postpartum sterilisation? Retrospective study on comparison between Pomeroy procedure and Filshie clips for a tubal occlusion at the time of Caesarean section. AB - OBJECTIVE: To compare the failure rate (pregnancies) of a Pomeroy procedure and Filshie clips tubal occlusion at the time of Caesarean section. METHOD: This is a retrospective observational study done in a district general hospital in the UK. There were 290 sterilisations performed at the time of Caesarean section over the period of 1994-2007. Studied population included 203 Pomeroy procedures and 87 Filshie clips applications. Follow-up period ranged from 2 to 15 years. A birth register and an operating theatre database were used to identify patients who underwent Caesarean section with a tubal occlusion. These patients' names were checked against the antenatal booking database, the early pregnancy assessment unit database, the operating theatre database in case of ectopic pregnancies, and a termination of pregnancy database to recognise failed sterilisation. RESULTS: There was no failure of tubal occlusion with a Pomeroy procedure (0/203). The failure rate of Filshie clips tubal occlusion was 1.15% (1/87) (p = 0.3). The length of the follow-up period ranged from 2 to 15 years (for Pomeroy procedure, median was 9 years and inter-quartile range (IQR) was 7; for Filshie clip, median was 8 years and IQR was 7). CONCLUSION: Pomeroy technique appears to carry a lower risk of a failed sterilisation than Filshie clips tubal occlusion at the time of Caesarean section. However, Pomeroy procedure needs to be balanced against the speed and simplicity of Filshie clips tubal occlusion. PMID- 20012306 TI - Ethnic differences in age-related changes of postpartum hemoglobin levels. AB - OBJECTIVE: Postpartum hemoglobin (Hb) levels may impact maternal and breast-fed newborn well being and health. Yet, the sources of variation in maternal postpartum Hb levels are poorly understood. We set to investigate whether age has an effect on postpartum Hb levels, and if this effect is similar in different ethnic groups. STUDY DESIGN: The birth records of over 200,000 births in a tertiary medical center were scanned for the first and second births to Jewish and Bedouin mothers in Israel. Age trends in Hb levels were analyzed using linear and bi-linear models. Confounding factors were eliminated by testing the significance of the preferred model in homogenous sub-populations. RESULTS: In Jewish Israeli mothers, postpartum hemoglobin level follow a bi-linear trend, peaking at the age of 27-31 years, both during the first and second births. In Israeli Bedouin mothers, a linear decline is observed through all ages. CONCLUSION: These results suggest an ethnic difference in the age trends of postpartum Hb levels. Clinically, significant changes may occur between the ages of 27 and 30 years. PMID- 20012305 TI - Inhibin/activin-betaC and -betaE subunits in the Ishikawa human endometrial adenocarcinoma cell line. AB - BACKGROUND: Inhibins and activins are important regulators of the female reproductive system. Recently, two novel inhibin subunits, named betaC and betaE, have been identified and shown to be expressed in several human tissues. However, only limited data on the expression of these novel inhibin subunits in normal human endometrial tissue and endometrial adenocarcinoma cell lines exist. MATERIALS AND METHODS: Samples of proliferative and secretory human endometrium were obtained from five premenopausal, non-pregnant patients undergoing gynecological surgery for benign diseases. Normal endometrial tissue and Ishikawa endometrial adenocarcinoma cell lines were analyzed by immunohistochemistry, immunofluorescence and RT-PCR. RESULTS: Expression of the inhibin betaC and betaE subunits could be demonstrated at the protein level by means of immunohistochemical evaluation and at the transcriptional level by establishing a betaC- and betaE-specific RT-PCR analysis in normal human endometrial tissue and the parental Ishikawa cell line. Interestingly, in a highly de-differentiated subclone of the Ishikawa cell line lacking estrogen receptor expression, the expression of the inhibin-betaC subunit appeared strongly reduced. DISCUSSION: Here, we show for the first time that the novel inhibin/activin-betaC and -betaE subunits are expressed in normal human endometrium and the estrogen receptor positive human endometrial carcinoma cell line Ishikawa using RT-PCR and immunohistochemical detection methods. Interestingly, the Ishikawa minus cell line (lacking estrogen receptor expression) demonstrated no to minimal expression of the betaC subunit as observed with immunofluorescence and RT-PCR, suggesting a possible hormone- dependency of this subunit in human endometrial cancer cells. Moreover, because the Ishikawa cell line minus is thought to be a more malignant endometrial cell line than its estrogen receptor positive counterpart, inhibin betaC subunit might be substantially involved in the pathogenesis and malignant transformation in human endometrium. PMID- 20012307 TI - No association of the G972S polymorphism of the insulin receptor substrate-1 gene with polycystic ovary syndrome in lean PCOS women with biochemical hyperandrogenemia. AB - PURPOSE: The aim of the present study was to determine the prevalence and association of the G972S polymorphism of the insulin receptor substrate-1 gene (IRS-1 G972S SNP) with polycystic ovary syndrome (PCOS) and insulin resistance related traits in a distinct phenotypic group of lean PCOS women with biochemical hyperandrogenemia, excluding obesity, which is considered to be an aggravating parameter of insulin resistance. METHODS: The study included 162 women with PCOS and 122 regularly menstruating, ovulatory women as controls. Physical measurements included weight, height, fat-free mass, fat mass, systolic and diastolic blood pressure and resting heart rate. Biochemical parameters included the serum testosterone, free testosterone, androstenedione, total cholesterol, triglycerides, HDL and LDL cholesterol and glucose levels. Insulin resistance was assessed by determining fasting insulin levels, fasting glucose levels, the fasting glucose/insulin ratio, as well as the HOMA and QUICKI indexes. All DNA samples were genotyped by a PCR-restriction fragment length polymorphism (RLFP) assay. RESULTS: No association of the genotype frequencies of the G972S polymorphism in insulin receptor substrate-1 gene (IRS-1 G972S SNP) with PCOS phenotype and insulin resistance was detected. CONCLUSION: The G972S polymorphism of the IRS-1 gene should not be viewed as major contributor to the development of PCOS or as a causative variant for insulin resistance. PMID- 20012308 TI - Randomized, controlled study of the effects of raloxifene on high sensitivity C reactive protein and serum lipids. AB - OBJECTIVE: To investigate the effects of raloxifene, on serum lipids and high sensitivity C-reactive protein (hs-CRP) in healthy postmenopausal women. METHODS: We studied the effect of raloxifene, on serum lipids and hs-CRP in 85 healthy postmenopausal women. Participants were randomly assigned to 60 mg daily raloxifene (43 subjects) for 6 months; the rest of the subjects were in the control group. Serum total cholesterol (TC), low-density lipoprotein cholesterol (LDL-C), high-density lipoprotein cholesterol, triglyceride and hs-CRP levels were measured at baseline and at the sixth month in both groups. RESULTS: Raloxifene treatment resulted in a 26% reduction in serum hs-CRP concentrations at the sixth month, compared with the baseline levels (P < 0.05). At the sixth month, TC and LDL-C levels were significantly reduced by 60 mg daily raloxifene (6.8 and 5.6%, respectively) when compared with both the baseline levels and the control group. CONCLUSION: The results of our study showed that raloxifene at a dose of 60 mg daily reduces serum TC, LDL-C and hs-CRP levels significantly in healthy postmenopausal women. According to the results of the current study, we suggest that raloxifene may have a favorable effect on the prevention of cardiovascular disease in healthy postmenopausal women. PMID- 20012309 TI - Prevalence of MS in Iranian immigrants to British Columbia, Canada. PMID- 20012310 TI - Dissociation between the severity of cerebrospinal fluid hypovolemia and MRI findings. PMID- 20012311 TI - Posterior reversible encephalopathy syndrome in a patient with acute intermittent porphyria. PMID- 20012312 TI - Autosomal dominant nemaline myopathy caused by a novel alpha-tropomyosin 3 mutation. AB - Nemaline myopathy (NM) is a genetically and clinically heterogenous muscle disorder, which is myopathologically characterized by nemaline bodies. Mutations in six genes have been reported to cause NM: Nebulin (NEB Pelin 1999), alpha skeletal muscle actin (ACTA1 Nowak 1999), alpha-slow tropomyosin (TPM3 Laing 1995), beta-tropomyosin (TPM2 Donner 2002), slow troponin T (TNNT1 Johnston 2000) and cofilin 2 (CFL2 Agrawal 2007). The majority of cases are due to mutation in NEB and ACTA1. We report on the clinical, myopathological and muscle MRI findings in a German family with autosomal dominant NM due to a novel pathogenic TPM3 mutation (p.Ala156Thr). PMID- 20012313 TI - Phenotype genotype analysis in 15 patients presenting a congenital myasthenic syndrome due to mutations in DOK7. AB - Congenital myasthenic syndromes (CMSs) are a heterogeneous group of diseases caused by genetic defects affecting neuromuscular transmission. Mutations of DOK7 have recently been described in recessive forms of CMS. Dok-7 is a cytoplasmic post-synaptic protein co-activator of the muscle-specific receptor-tyrosine kinase (MuSK) involved in neuromuscular synaptogenesis and maintenance. We report clinical, morphological and molecular data on 15 patients with mutations in DOK7. Eleven different mutations (5 novel) were identified and all patients but one were found to carry at least the common c.1124_1127dupTGCC mutation. Patients with DOK7 mutations have a particular limb-girdle pattern, without tubular aggregates but a frequent lipidosis on the muscle biopsy. Changes in pre- and post-synaptic compartments of the neuromuscular junction were also observed in muscle biopsies: terminal axons showed defective branching which resulted in a unique terminal axon contacting en passant postsynaptic cups. Clinical features, muscle biopsy findings or response to therapy were confusing in several patients. Characterization of this distinct phenotype is essential to provide clues for targeted genetic screening and to predict the therapeutic response to anticholinesterase treatments or ephedrine as has been suggested. PMID- 20012314 TI - Acute transverse myelitis in psoriatic arthritis. PMID- 20012315 TI - Effects of carbon dioxide pneumoperitoneum on hepatic function in obstructive jaundice: an experimental study in a rat model. AB - BACKGROUND AND AIMS: The physiology of the patient during laparoscopy differs from that of open surgery. Both pneumoperitoneum and obstructive jaundice impair the hepatic function, but the combined insult has not been previously examined. In this study, we aimed to investigate the effects of carbon dioxide (CO(2)) pneumoperitoneum on hepatic function in a rat model of obstructive jaundice. METHODS: Forty-four male Sprague-Dawley rats were divided into four groups: group 1 (n = 10), sham-operated group; group 2 (n = 12), obstructive jaundice group; group 3 (n = 10), CO(2) pneumoperitoneum group; and group 4 (n = 12), obstructive jaundice and CO(2) pneumoperitoneum group. Common bile duct was ligated and divided in the obstructive jaundice groups. After 6 days, a 12-mmHg pneumoperitoneum was induced, maintained for 60 min, and released for 120 min. Blood samples were drawn for the measurement of white blood cell and platelet counts, serum liver enzymes (aspartate aminotransferase [AST], alanine aminotransferase [ALT], total bilirubin). Tissue samples were obtained for analyses of malondialdehyde (MDA), glutathione (GSH), and superoxide dismutase (SOD) levels. We evaluated the degree of liver injury on a grading scale from 0 to 4, histopathologically. RESULTS: Pneumoperitoneum after biliary obstruction resulted in an increase in AST and ALT levels and a decrease in white blood cell and platelet counts. However, changes in liver tissue MDA, GSH, and SOD levels did not correlate with the changes in AST and ALT levels and white blood cell and platelet counts. After sham operation with pneumoperitoneum, the GSH levels in liver homogenate were significantly decreased in the group 3 when compared to the group 2. On the other hand, obstructive jaundice itself caused significant reduction in the SOD activity of liver homogenate in comparison to the group 3. Histopathologically, sinusoidal congestion and vacuolization were more severe in the group 3. CONCLUSIONS: Alterations in hepatic function occur in pneumoperitoneum applied jaundiced subjects. However, there were no statistically significant differences between the groups 2 and 4 with regard to white blood cell and platelet counts, serum liver enzymes including AST, ALT, and total bilirubin values, MDA and GSH levels and SOD activity of liver homogenate, and histologic damage. These results indicate that there is no additional risk on liver function associated with pneumoperitoneum performed in obstructive jaundice. PMID- 20012316 TI - The value of intraoperative frozen section examination of sentinel lymph nodes in surgical management of breast carcinoma. AB - PURPOSE: In breast cancer staging, the need for intraoperative sentinel lymph node (SLN) evaluation has still not been adequately established. This study investigates the impact of intraoperative frozen section (FS) evaluation of SLN to avoid subsequent axillary lymph node dissection (ALND) in patients with positive SLN. METHODS: A retrospective review of 364 breast cancer patients undergoing SLN biopsy with intraoperative FS evaluation of SLN was performed. RESULTS: Sensitivity and accuracy of FS examination of SLN were 76.4% and 94.2%, respectively. The sensitivity was significantly higher in larger tumors (p < 0.01). No significant correlation was found between FS and histologic type. A second intervention was avoided in 83.9% of the patients with positive SLN. CONCLUSIONS: Intraoperative FS examination of the SLN is a useful predictor of axillary lymph node status in breast cancer patients. The majority of SLN positive patients allows for immediate ALND in a one-stage surgical procedure. PMID- 20012317 TI - Expression of thymidylate synthase determines the response of gastric cancer patients undergoing gastrectomy to 5-fluorouracil-based adjuvant chemotherapy. AB - PURPOSE: We investigated whether the intensity of thymidylate synthase (TS) staining in tissue samples obtained from gastric cancer (GC) patients undergoing gastrectomy could predict response to 5-FU-based adjuvant chemotherapy after gastrectomy. METHOD AND MATERIALS: Clinicopathological features of 124 patients with histologically proven GC who underwent radical gastrectomy were retrospectively reviewed. Tissue samples obtained from these patients were immunohistochemically stained for assessing TS expression. We arbitrarily classified the TS staining results as low (<20% cytoplasmic immunostaining) and high (> or =20% cytoplasmic immunostaining) TS expression. RESULTS: The clinicopathological features of the low TS expression group patients were typically similar to those of the high TS expression group patients. However, multivariate forward stepwise logistic regression analysis revealed that low TS expression was independently associated with females and responders to 5-FU-based adjuvant chemotherapy. The median follow-up duration for the 124 GC patients who had undergone curative resection was 41.3 months. The GC patients who showed poor tumor differentiation and high TS expression had short disease-free survival (DFS) and overall survival (OS). CONCLUSIONS: Low TS expression is significantly associated with female GC patients and responders to 5-FU-based adjuvant chemotherapy. It predicts longer DFS and OS in selected GC patients treated with 5-FU-based adjuvant chemotherapy after curative resection. The results suggest that prospective assessment of TS staining intensity in tissue samples obtained from GC patients undergoing gastrectomy would be useful to predict the patients who would be benefited from 5-FU-based adjuvant chemotherapy after gastrectomy. PMID- 20012318 TI - The impact of the National Patient Safety Agency intravenous fluid alert on iatrogenic hyponatraemia in children. AB - In March 2007, the National Patient Safety Agency (NPSA) issued an alert regarding intravenous fluid (IVF) prescription to hospitalised infants and children, to be implemented in UK hospitals by September 2007. Previously, the most commonly used IVF (0.18% saline/4% dextrose) has been associated with iatrogenic hyponatraemia, resulting in four deaths and one near miss since 2000. The alert recommended 0.45% (or 0.9%) saline/5% dextrose as maintenance IVF and banned 0.18% saline/4% dextrose. We audited practice and outcome in children receiving maintenance IVF in June 2007 (before guideline implementation) and June 2008 (after guideline implementation). In June 2007, 44 (30%) children were prescribed IVF, six received IVF not recommended by NPSA alert 22 and one became hyponatraemic. In June 2008, 56 (30%) children received IVF; one received IVF not recommended by NPSA alert 22 and became hyponatraemic. The median change in serum sodium levels for all children who received IVF not recommended by NPSA alert 22 [-5 (-15 to 0) mmol/l] was significantly greater than those who received IVF recommended by NPSA alert 22 [0 (-13 to +7) mmol/l, p = 0.002]. In addition, there was a significant (p = 0.04) reduction in the number of children who had electrolytes checked while on IVF after implementation of the guideline. Implementation of a new IVF guideline has been associated with less use of IVF not recommended by NPSA alert 22, resulting in less serum sodium level reduction. The only children who became hyponatraemic received IVF not recommended by NPSA alert 22. Despite the NPSA alert and guideline implementation, less children had electrolyte levels checked while receiving IVF. PMID- 20012319 TI - Medullary neurons in the core white matter of the olfactory bulb: a new cell type. AB - The structure of a new cell type, termed the medullary neuron (MN) because of its intimate association with the rostral migratory stream (RMS) in the bulbar core, is described in the adult rat olfactory bulb. The MN is a triangular or polygonal interneuron whose soma lies between the cellular clusters of the RMS or, less frequently, among the neuron progenitors therein. MNs are easily distinguished from adjacent cells by their large size and differentiated structure. Two MN subtypes have been categorized by the Golgi technique: spiny pyramidal neurons and aspiny neurons. Both MN subtypes bear a large dendritic field impinged upon by axons in the core bulbar white matter. A set of collaterals from the adjacent axons appears to terminate on the MN dendrites. The MN axon passes in close apposition to adjacent neuron progenitors in the RMS. MNs are immunoreactive with antisera raised against gamma-aminobutyric acid and glutamate decarboxylase 65/67. Electron-microscopic observations confirm that MNs correspond to fully differentiated, mature neurons. MNs seem to be highly conserved among macrosmatic species as they occur in Nissl-stained brain sections from mouse, guinea pig, and hedgehog. Although the functional role of MNs remains to be determined, we suggest that MNs represent a cellular interface between endogenous olfactory activity and the differentiation of new neurons generated during adulthood. PMID- 20012320 TI - Silicatein-mediated incorporation of titanium into spicules from the demosponge Suberites domuncula. AB - Primmorphs (a three-dimensional sponge primary cell culture system) have been revealed to be a cell/tissue nano-factory for the production of tailor-made hybrid nanostructures. Growth of primmorphs is stimulated by the presence of a titanium alkoxide precursor tolerating titania (TiO2) concentrations up to 250 microM. The presence and activity of silicatein in primmorphs has been analyzed by gel electrophoresis and Western blotting. Results of studies by scanning transmission electron microscopy with energy-dispersive X-ray spectroscopy have revealed silica and titania to be co-localized on nanosized spicules. Our findings suggest that the incorporation of titania into the nanosized spicule is enzymatically mediated via active silicatein in an orchestrated mechanism. PMID- 20012321 TI - Expression of the actin-associated protein transgelin (SM22) is decreased in prostate cancer. AB - Transgelin is an actin-binding protein shown to be tumour-suppressive. Loss of transgelin expression in transformed cells is associated with oncogenesis. This study aimed to determine whether transgelin expression was suppressed in prostate cancer. An in silico meta-analysis with public-domain expressed-sequence-tag libraries of normal human prostate epithelium, prostatic intraepithelial neoplasia, invasive carcinoma and metastasised lesions predicted decreased transgelin expression with disease progression. Similarly, analysis of Affymetrix gene chip data and the Oncomine database indicated that transgelin was one the 2% most significant of all down-regulated genes in response to prostate cancer. Analysis by quantitative reverse transcription with the polymerase chain reaction (qRT-PCR) of patient biopsies determined transgelin expression to be significantly lower in prostate tumour tissue than in matched normal tissue. Similarly, qRT-PCR and Western blot analysis of representative prostate cancer cell lines demonstrated significantly lower levels of transgelin mRNA and protein in all but the DU145 prostate cancer cell line. Increased expression of TAGLN and increased transgelin protein in response to treatment with transforming growth factor-beta suggested that reduced expression in prostate cancer was not attributable to gene promoter suppression by hypermethylation. Gene ontology function analysis highlighted the importance of transgelin in the co-deregulation of actin-binding proteins. Thus, transgelin is suppressed during prostate cancer progression and seems to be an important factor in the dysregulation of the actin cytoskeleton. PMID- 20012322 TI - The subcommissural organ and the development of the posterior commissure in chick embryos. AB - The subcommissural organ (SCO) is an ependymal differentiation located in the diencephalon under the posterior commissure (PC). SCO-spondin, a glycoprotein released by the SCO, belongs to the thrombospondin superfamily and shares molecular domains with axonal pathfinding molecules. Several lines of evidence suggest a relationship between the SCO and the development of the PC in the chick: (1) their close location to each other, (2) their differentiation at the same developmental stage in the chick, (3) the abnormal PC found in null mutants lacking an SCO and (4) the release by the SCO of SCO-spondin. By application of DiI crystals in the PC of chick embryos, we have identified the neurons that give rise to the PC. Labelling is confined to the magnocellular nucleus of the PC (MNPC). To gain insight into the role of the SCO in PC development, coculture experiments of explants of the MNPC region (MNPCr) from embryos at embryonic day 4 (E4) with SCO explants from E4 or E13 embryos have been performed and the neurite outgrowth from the MNPCr explants has been analysed. In the case of coculture of E4 MNPCr with E4 SCO, the number of neurites growing from the MNPCr is higher at the side facing the SCO. However, when E4 MNPCr and E13 SCO are cocultured, the neurites grow mostly at the side opposite to the SCO. These data suggest that, at early stages of development, the SCO releases some attractive or permissive molecule(s) for the growing of the PC, whereas at later stages, the SCO has a repulsive effect over neurites arising from MNPCr. PMID- 20012323 TI - Post-ERCP pancreatitis. AB - Pancreatitis remains the most common severe complication of endoscopic retrograde cholangiopancreatography (ERCP). Detailed information about the findings of previous studies concerning post-ERCP pancreatitis has not been utilized sufficiently. The purpose of the present article was to present guidelines for the diagnostic criteria of post-ERCP pancreatitis, and its incidence, risk factors, and prophylactic procedures that are supported by evidence. To achieve this purpose, a critical examination was made of the articles on post-ERCP pancreatitis, based on the data obtained by research studies published up to 2009. At present, there are no standardized diagnostic criteria for post-ERCP pancreatitis. It is appropriate that post-ERCP pancreatitis is defined as acute pancreatitis that has developed following ERCP, and its diagnosis and severity assessment should be made according to the diagnostic criteria and severity assessment of the Japanese Ministry of Health, Labour and Welfare. The incidence of acute pancreatitis associated with diagnostic and therapeutic ERCP is 0.4-1.5 and 1.6-5.4%, respectively. Endoscopic papillary balloon dilation is associated with a high risk of acute pancreatitis compared with endoscopic sphincterotomy. It was made clear that important risk factors include dysfunction of the Oddi sphincter, being of the female sex, past history of post-ERCP pancreatitis, and performance of pancreaticography. Temporary prophylactic placement of pancreatic stents in the high-risk group is useful for the prevention of post-ERCP pancreatitis [odds ratio (OR) 3.2, 95% confidence interval (CI) 1.6-6.4, number needed to treat (NNT) 10]. Use of nonsteroidal anti-inflammatory drugs (NSAIDs) is associated with a reduction in the development of post-ERCP pancreatitis (OR 0.46, 95% CI 0.32-0.65). Single rectal administration of NSAIDs is useful for the prevention of post-ERCP pancreatitis [relative risk (RR) 0.36, 95% CI 0.22-0.60, NNT 15] and decreases the development of pancreatitis in both the low-risk group (RR 0.29, 95% CI 0.12-0.71) and the high-risk group (RR 0.40, 95% CI 0.23-0.72) of post-ERCP pancreatitis. As for somatostatin, a bolus injection may be most useful compared with short- or long-term infusion (OR 0.271, 95% CI 0.138-0.536, risk difference 8.2%, 95% CI 4.4-12.0%). The usefulness of gabexate mesilate was not apparent in any of the following conditions: acute pancreatitis (control 5.7 vs. 4.8% for gabexate mesilate), hyperamylasemia (40.6 vs. 36.9%), and abdominal pain (1.7 vs. 8.9%). Formulation of diagnostic criteria for post-ERCP pancreatitis is needed. Temporary prophylactic placement of pancreatic stents in the high-risk group offers the most promise as a means of preventing post-ERCP pancreatitis. As for pharmacological attempts, there are high expectations concerning NSAIDs because they are excellent in terms of cost-effectiveness, ease of use, and safety. There was no evidence of effective prophylaxis with the use of protease inhibitors, especially gabexate mesilate. PMID- 20012324 TI - Pancreatitis bundles. AB - Clinical indicators set forth in the guidelines have been found to contribute to the improvement in compliance with the guidelines. On the other hand, it has been shown that clinical indicators are more effective when individual indicators are presented in the form of a bundle than when they are given separately. Accordingly, in the JPN Guidelines 2010 for management of acute pancreatitis, those indicators that are judged to be important on the basis of a recommendation classification of "A or B" are presented as a pancreatitis bundle. Each item includes assessment of severity after a diagnosis of pancreatitis has been made, differentiation of pathogenesis, management of gallstone-induced pancreatitis, a sufficient dose of fluid replacement and monitoring, pain control, prophylactic administration of wide-spectrum antibiotics and cholecystectomy following resolution of pancreatic symptoms caused by cholecystolithiasis. Hereafter, the efficacy of these indicators and the significance of their achievement should be examined carefully. Then, the assessment of the compliance rate with the guidelines as well as the assessment of the guidelines and pancreatitis itself should become possible. PMID- 20012325 TI - Treatment strategy for acute pancreatitis. AB - When a diagnosis of acute pancreatitis (AP) is made, fundamental medical treatment consisting of fasting, intravenous (IV) fluid replacement, and analgesics with a close monitoring of vital signs should be immediately started. In parallel with fundamental medical treatment, assessment of severity based on clinical signs, blood test, urinalysis and imaging tests should be performed to determine the way of treatment for each patient. A repeat evaluation of severity is important since the condition is unstable especially in the early stage of AP. At the time of initial diagnosis, the etiology should be investigated by means of blood test, urinalysis and diagnostic imaging. If a biliary pancreatitis accompanied with acute cholangitis or biliary stasis is diagnosed or suspected, an early endoscopic retrograde cholangiopancreatography with or without endoscopic sphincterotomy (ERCP/ES) is recommended in addition to the fundamental medical treatment. In mild cases, the fundamental medical treatment should be continued until clinical symptom is subsided with normal laboratory data. In cases with severe acute pancreatitis (SAP) referral should be considered to medical centers experienced in the treatment of SAP, and intensive care is recommended for preventing both organ failures and infectious complications. Hemodynamic stabilization with vigorous fluid resuscitation, respiratory support and antibiotics are the major parts of intensive care in the early period of SAP. Continuous hemodiafiltration (CHDF) and continuous regional arterial infusion (CRAI) of protease inhibitor and/or antibiotics may be effective to improve pathophysiology of AP especially in the early stage of the disease. In the late stage of AP, infectious complications are critical. If an infectious complication is suspected based on clinical signs, blood test and imaging, a fine needle aspiration (FNA) is recommended to establish a diagnosis. The accuracy of FNA is reported to be 89 ~ 100%. For patients with sterile pancreatitis, non-surgical treatment should be indicated. For patients with infected pancreatic necrosis, therapeutic intervention either by percutaneous, endoscopic, laparoscopic or surgical approach are indicated. The most preferred surgical intervention is necrosectomy, however, non-surgical treatment with antibiotics is still the treatment of choice if the general condition is stable. Necrosectomy should be performed as late as possible. For patients with pancreatic abscess, drainage is recommended. PMID- 20012326 TI - Gallstone-induced acute pancreatitis. AB - In the care of acute pancreatitis, a prompt search for the etiologic condition of the disease should be conducted. A differentiation of gallstone-induced acute pancreatitis should be given top priority in its etiologic diagnosis because it is related to the decision of treatment policy. Examinations necessary for diagnosing gallstone-induced acute pancreatitis include blood tests and ultrasonography. Early ERCP/ES should be performed in patients with gallstone induced acute pancreatitis if a complication of cholangitis and a prolonged passage disorder of the biliary tract are suspected. The treatment for bile duct stones with the use of ERCP/ES alone is not recommended in cases of gallstone induced pancreatitis with gallbladder stones. Cholecystectomy for gallstone induced acute pancreatitis should be performed using a laparoscopic procedure as the first option as soon as the disease has subsided. PMID- 20012327 TI - Health insurance and payment systems for severe acute pancreatitis. AB - The medical insurance system of Japan is based on the Universal Medical Care System guaranteed by the provision of the Article 25 of the Constitution of Japan, which states that "All the people shall have the right to live a healthy, cultural and minimum standard of life." The health insurance system of Japan comprises the medical insurance system and the health care system for the long lived. Medical care insurance includes the employees' health insurance (Social Insurance) that covers employees of private companies and their families and community insurance (National Health Insurance) that covers the self-employed. Each medical insurance system has its own medical care system for the retired and their families. The health care system for the long-lived covers people of over 75 years of age (over 65 years in people with a certain handicap). There is also a system under which all or part of the medical expenses is reimbursed by public expenditure or the cost of medical care not covered by health insurance is paid by the government. This system is referred to collectively as the "the public payment system of medical expenses." To support the realization of the purpose of this system, there is a treatment research enterprise for specified diseases (intractable diseases). Because of the high mortality rate, acute pancreatitis is specified as an intractable disease for the purpose of reducing its mortality rate, and treatment expenses of patients are paid in full by the government dating back to the day when the application was made for a certificate verifying that he or she has severe acute pancreatitis. PMID- 20012328 TI - New diagnostic criteria of acute pancreatitis. AB - Practical guidelines for the diagnosis of acute pancreatitis are presented so that a rapid and adequate diagnosis can be made. When acute pancreatitis is suspected in patients with acute onset of abdominal pain and tenderness mainly in the upper abdomen, the diagnosis of acute pancreatitis is made on the basis of elevated levels of pancreatic enzymes in the blood and/or urine. Furthermore, other acute abdominal diseases are ruled out if local findings associated with pancreatitis are confirmed by diagnostic imaging. According to the diagnostic criteria established in Japan, patients who present with two of the following three manifestations are diagnosed as having acute pancreatitis: characteristic upper abdominal pain, elevated levels of pancreatic enzymes, and findings of ultrasonography (US), CT or MRI suggesting acute pancreatitis. Detection of elevated levels of blood pancreatic enzymes is crucial in the diagnosis of acute pancreatitis. Measurement of blood lipase is recommended, because it is reported to be superior to all other pancreatic enzymes in terms of sensitivity and specificity. For measurements of the blood amylase level widely used in Japan, it should be cautioned that, because of its low specificity, abnormal high values are also often obtained in diseases other than pancreatitis. The cut-off level of blood pancreatic enzymes for the diagnosis of acute pancreatitis is not able to be set because of lack of sufficient evidence and consensus to date. CT study is the most appropriate procedure to confirm image findings of acute pancreatitis. Elucidation of the etiology of acute pancreatitis should be continued after a diagnosis of acute pancreatitis. In the process of the etiologic elucidation of acute pancreatitis, judgment whether it is gallstone-induced or not is most urgent and crucial for deciding treatment policy including the assessment of whether endoscopic papillary treatment should be conducted or not. The diagnosis of gallstone-induced acute pancreatitis can be made by combining detection of elevated levels of bilirubin, transamylase (ALT, AST) and ALP detected by hematological examination and the visualization of gallstones by US. PMID- 20012329 TI - Assessment of severity of acute pancreatitis according to new prognostic factors and CT grading. AB - The assessment of severity at the initial medical examination plays an important role in introducing adequate early treatment and the transfer of patients to a medical facility that can cope with severe acute pancreatitis. Under these circumstances, "criteria for severity assessment" have been prepared in various countries, including Japan, and these criteria are now being evaluated. The criteria for severity assessment of acute pancreatitis in Japan were determined in 1990 (of which a partial revision was made in 1999). In 2008, an overall revision was made and the new Japanese criteria for severity assessment of acute pancreatitis were prepared. In the new criteria for severity assessment, the diagnosis of severe acute pancreatitis can be made according to 9 prognostic factors and/or the computed tomography (CT) grades based on contrast-enhanced CT. Patients with severe acute pancreatitis are expected to be transferred to a specialist medical center or to an intensive care unit to receive adequate treatment there. In Japan, severe acute pancreatitis is recognized as being a specified intractable disease on the basis of these criteria, so medical expenses associated with severe acute pancreatitis are covered by Government payment. PMID- 20012330 TI - Decompression of a pneumoperitoneum during an extraperitoneal inguinal hernia repair (TEP) by incision of the umbilical sac under direct vision. PMID- 20012331 TI - Risk factors for bowel resection and outcome in patients with incarcerated groin hernias. AB - PURPOSE: This retrospective study aimed to evaluate the risk factors for bowel resection and outcome in patients with incarcerated groin hernias. METHODS: The records of all adult patients who had undergone emergency hernia repair for incarcerated groin hernia from January 1999 to June 2009 were analyzed. One hundred and eighty-two patients with incarcerated groin hernias were included in this study. Bowel resection was required in 28 patients (15.4%). RESULTS: One hundred and twenty-six patients (69.2%) were covered by various types of health insurance. Twenty-six patients (14.3%) were hospitalized for mechanical bowel obstruction or had obvious symptoms of simultaneous bowel obstruction. Obvious peritonitis was found in seven patients (3.8%). A logistic regression model identified three independent risk factors for bowel resection: lack of health insurance (odds ratio [OR], 5, P = 0.005), obvious peritonitis (OR, 11.52, P = 0.019), and femoral hernia (OR, 8.31, P < 0.001). Postoperative complications (standardized coefficient [SC] = 0.478, P < 0.001), presentation of bowel obstruction on admission (SC = 0.169, P = 0.017), and having health insurance (SC = 0.153, P = 0.030) were associated with prolonged length of stay. No factors were found to be directly associated with morbidity or mortality. CONCLUSIONS: Risk for intestinal resection in emergency groin hernia repair is higher in patients with femoral hernia, obvious peritonitis, or those with no health insurance. Surgeons should pay more attention to these patients and initiate emergency surgery without delay. PMID- 20012332 TI - Fatal necrotizing fasciitis following elective inguinal hernia repair. AB - Necrotizing soft tissue infections (NSTIs) following elective hernia repair are extremely uncommon, though they can occur following emergency surgery for complicated hernias. They are also usually seen in individuals with impaired immunity. We report a case of fatal necrotizing fasciitis following elective hernia repair in an otherwise healthy young patient. A high index of suspicion is required to diagnose this condition early, as it is difficult to differentiate it from superficial surgical site infection. PMID- 20012333 TI - Hernia repair: the search for ideal meshes. AB - BACKGROUND: Effective repair of hernia is a difficult task. There have been many advances in hernia repair techniques over the past 50 years, but new strategies must be considered to enhance the success of herniorrhaphy. DISCUSSION: At the 30th International Congress of the European Hernia Society, nine experts in hernia repair and experimental mesh evaluation participated in a roundtable discussion about today's unmet needs in hernia repair, including what constitutes an "ideal" hernia repair and the portfolio of "ideal" mesh prostheses. Defining characteristics of lightweight mesh, mesh alternatives, the surgeon's role in hernia repair, adverse events, the unmet requirements for today's hernia repair, and optimized animal models were among the topics discussed. CONCLUSION: The ideal mesh's construction is still in progress, but greater understanding of its critical characteristics was explored. It is hoped that these suggestions will lead to the development of improved hernia treatments and a maximally effective portfolio of hernia mesh prostheses. PMID- 20012335 TI - Bioleaching of chalcopyrite concentrate using Leptospirillum ferriphilum, Acidithiobacillus ferrooxidans and Acidithiobacillus thiooxidans in a continuous bubble column reactor. AB - To estimate the bioleaching performance of chalcopyrite for various hydraulic residence times (HRTs), laboratory-scale bioleaching of chalcopyrite concentrate was carried out in a continuous bubble column reactor with three different HRTs of 120, 80 and 40 h, respectively. An extraction rate and ratio of 0.578 g Cu l( 1) h(-1) and 39.7%, respectively, were achieved for an HRT of 80 h at a solids concentration of 10% (w/v). Lower bioleaching performances than this were obtained for a longer HRT of 120 h and a shorter HRT of 40 h. In addition, there was obvious competition between Leptospirillum ferriphilum and Acidithiobacillus ferrooxidans to oxidize ferrous iron, causing large compositional differences between the microbial communities obtained for the different HRTs. Leptospirillum ferriphilum and Acidithiobacillus thiooxidans were found to be the dominant microbes for the longer HRT (120 h). Acidithiobacillus ferrooxidans became the dominant species when the HRT was decreased. The proportion of Acidithiobacillus thiooxidans was comparatively constant in the microbial community throughout the three process stages. PMID- 20012334 TI - Six weeks antibiotic therapy for all bone infections: results of a cohort study. AB - There is no consensus on the antibiotic therapy for bone infection due to the heterogeneous spectrum of diseases. Most authors suggest different durations of treatment based on pathophysiological considerations. However, adverse effects are related, at least in part, to the duration of treatment. We, therefore, investigated a 6 weeks antibiotic combination therapy for all cases of bone infection. Herewith, we report the results of this therapeutic approach. This is a cohort study including all patients presenting with bone infection, regardless of the mechanism involved. The diagnosis was based on bone biopsy obtained through invasive procedures. Chronic bone infection was defined as a history of disease of over 1 month duration. The duration of clinical follow-up following treatment discontinuation was at least 6 months. Cured bone infection was defined as the absence of relapse after antibiotic discontinuation. One hundred and eighteen patients were included between July 2005 and March 2009; 61 presented with bone infection following prosthetic implant (52%) and the 57 remaining patients had bone infection without foreign material (48%). Surgery was required for 80 patients (68%). Microbial agents were identified in 116/118 patients, with 24 patients presenting with polymicrobial sepsis (20%). The mean duration of antibiotic treatment was 42 +/- 0.2 days and the mean clinical follow-up was 27 +/- 14 months. The treatment success rate was 91.5% (108/118). Six weeks of antimicrobial therapy appears to be effective for nearly all bone infections, regardless of the pathophysiology. These results encourage us to pursue attempts to simplify the management of bone infection without obvious prejudice to the patient. PMID- 20012336 TI - Anti-tumor activity and mechanisms of a novel vascular disrupting agent, (Z) 3,4',5-trimethoxylstilbene-3'-O-phosphate disodium (M410). AB - Vascular disrupting agents (VDAs) have emerged as a new kind of anti-cancer drug in recent years. Structural modification of an active parent compound is an effective approach to developing new agents with more activity and fewer adverse reactions. In our study, six synthesized stilbene derivatives were screened for their cytotoxic activity against human tumor cells, and their mechanisms of action were investigated. The MTT assay was used to determine the anti proliferative activity of these compounds. Polymerization of tubulin was detected by a tubulin assembly assay, and the cellular microtubule network was observed by immunocytochemical analyses. Cell-cycle distribution was detected by flow cytometry. A nude mouse model with xenografted colon cancer was used to demonstrate the in vivo anti-tumor activity, and microvessel density (MVD) was determined by immunohistochemistry. The expression levels of protein and mRNA were detected by Western blot and RT-PCR, respectively. Among the six newly synthesized compounds, (Z)-3,4',5-trimethoxylstilbene-3'-O-phosphate disodium (M410) showed potent cytotoxic activity toward proliferating tumor cells and exhibited a similar cytotoxicity against multi-drug resistant (MDR) tumor cells. M410 inhibited bovine brain tubulin polymerization in a way similar to that of colchicine. In proliferating human umbilical vein endothelial cells (HUVECs), 20 nM of M410 induced cellular tubulin depolymerization within 4 h, which led to M phase arrest. Systemic administration of M410 at nontoxic doses in nude mice resulted in inhibition of tumor growth of human colon cancer LoVo xenografts. The tumor vessel density also decreased after M410 treatment, as determined by immunohistochemical staining for CD31. M410 downregulated hypoxia-inducible factor-1 alpha (HIF-1alpha) expression, reduced nuclear HIF-1alpha, and downregulated vascular endothelial cell growth factor (VEGF) mRNA. Our results indicate that M410 is a potent microtubule inhibitor that is cytotoxic, angiogenesis inhibiting and vascular targeting. PMID- 20012337 TI - A phase II study of everolimus in combination with imatinib for previously treated advanced renal carcinoma. AB - PURPOSE: This phase II study evaluated the activity of combined treatment with the mTOR inhibitor everolimus and the PDGFR inhibitor imatinib in patients with previously-treated, advanced renal carcinoma. The primary endpoint was estimation of the 3-month progression-free rate. PATIENTS AND METHODS: Eligible patients had metastatic or unresectable clear cell renal carcinoma, at least one prior systemic therapy, no prior mTOR inhibitor therapy, performance status 0-2, and measurable disease. Treatment consisted of everolimus 2.5 mg p.o. daily and imatinib 600 mg p.o. daily. The primary endpoint was the 3-month progression-free rate. RESULTS: The study was closed after the first 19 patients because of an insufficient number of patients who were progression-free at 3 months. The 3 month progression-free rate was 49% (95% C.I. 23%, 72%) and the median progression-free survival was 2.9 months (95% C.I. 1.9, 6.2). Toxicities with an incidence of > 50% included nausea, elevated serum creatinine, edema, anemia, hypocalcemia, fatigue, diarrhea, vomiting, and dyspnea, and leukopenia. CONCLUSION: The combination of everolimus with imatinib in previously treated patients with advanced renal carcinoma did not result in a sufficient 3-month progression-free rate to warrant further investigation of this combination. PMID- 20012338 TI - Dibutyltin(IV) complexes containing arylazobenzoate ligands: chemistry, in vitro cytotoxic effects on human tumor cell lines and mode of interaction with some enzymes. AB - Dibutyltin(IV) complexes of composition Bu2Sn(LH)2, where LH is a carboxylate residue derived from 2-[(E)-(5-tert-butyl-2-hydroxyphenyl)diazenyl]benzoate (L1H) with water molecule (1), 4-[(E)-(5-tert-butyl-2-hydroxyphenyl)diazenyl]benzoate (L2H) (2) and 4-[(E)-(4-hydroxy-5-methylphenyl)diazenyl]benzoate (L3H) (3), were synthesized and characterized by spectroscopic (1H, 13C and 119Sn NMR, IR, 119Sn Mossbauer) techniques. A full characterization was accomplished from the crystal structure of complex 1. The molecular structures and geometries of the complexes (1a i.e. 1 without water molecule and 3) were fully optimized using the quantum mechanical method (PM6). Complexes 1 and 3 were found to exhibit stronger cytotoxic activity in vitro across a panel of human tumor cell lines viz., A498, EVSA-T, H226, IGROV, M19 MEL, MCF-7 and WIDR. Compound 3 is found to be four times superior for the A498, EVSA-T and MCF-7 cell lines than CCDP (cisplatin), and four, eight and sixteen times superior for the A498, H226 and MCF-7 cell lines, respectively, compared to ETO (etoposide). The mechanistic role of cytotoxic activity of test compounds is discussed in relation to the theoretical results of docking studies with some key enzymes such as ribonucleotide reductase, thymidylate synthase, thymidylate phosphorylase and topoisomerase II associated with the propagation of cancer. PMID- 20012339 TI - Herman Skolnik award symposium honoring Yvonne Martin. PMID- 20012340 TI - Anaerobic digestion of solid slaughterhouse waste: study of biological stabilization by Fourier Transform infrared spectroscopy and thermogravimetry combined with mass spectrometry. AB - In this paper, Fourier Transform infrared spectroscopy (FTIR) along with thermogravimetric analysis together with mass spectrometry (TG-MS analysis) were employed to study the organic matter transformation attained under anaerobic digestion of slaughterhouse waste and to establish the stability of the digestates obtained when compared with fresh wastes. Digestate samples studied were obtained from successful digestion and failed systems treating slaughterhouse waste and the organic fraction of municipal solid wastes. The FTIR spectra and TG profiles from well stabilized products (from successful digestion systems) showed an increase in the aromaticity degree and the reduction of volatile content and aliphatic structures as stabilization proceeded. On the other hand, the FTIR spectra of non-stable reactors showed a high aliphaticity degree and fat content. When comparing differential thermogravimetry (DTG) profiles of the feed and digestate samples obtained from all successful anaerobic systems, a reduction in the intensity of the low-temperature range (approximately 300 degrees C) peak was observed, while the weight loss experienced at high temperature (450-550 degrees C) was variable for the different systems. Compared to the original waste, the intensity of the weight loss peak in the high temperature range decreased in the reactors with higher hydraulic retention time (HRT) whereas its intensity increased and the peak was displaced to higher temperatures for the digesters with lower HRT. PMID- 20012341 TI - Effects of gasoline components on MTBE and TBA cometabolism by Mycobacterium austroafricanum JOB5. AB - In this study we have examined the effects of individual gasoline hydrocarbons (C(5-10,12,14) n-alkanes, C(5-8) isoalkanes, alicyclics [cyclopentane and methylcyclopentane] and BTEX compounds [benzene, toluene, ethylbenzene, m-, o-, and p-xylene]) on cometabolism of methyl tertiary butyl ether (MTBE) and tertiary butyl alcohol (TBA) by Mycobacterium austroafricanum JOB5. All of the alkanes tested supported growth and both MTBE and TBA oxidation. Growth on C(5-8) n alkanes and isoalkanes was inhibited by acetylene whereas growth on longer chain n-alkanes was largely unaffected by this gas. However, oxidation of both MTBE and TBA by resting cells was consistently inhibited by acetylene, irrespective of the alkane used as growth-supporting substrate. A model involving two separate but co expressed alkane-oxidizing enzyme systems is proposed to account for these observations. Cyclopentane, methylcyclopentane, benzene and ethylbenzene did not support growth but these compounds all inhibited MTBE and TBA oxidation by alkane grown cells. In the case of benzene, the inhibition was shown to be due to competitive interactions with both MTBE and TBA. Several aromatic compounds (p xylene > toluene > m-xylene) did support growth and cells previously grown on these substrates also oxidized MTBE and TBA. Low concentrations of toluene (<10 microM) stimulated MTBE and TBA oxidation by alkane-grown cells whereas higher concentrations were inhibitory. The effects of acetylene suggest strain JOB5 also has two distinct toluene-oxidizing activities. These results have been discussed in terms of their impact on our understanding of MTBE and TBA cometabolism and the enzymes involved in these processes in mycobacteria and other bacteria. PMID- 20012342 TI - Plant-eriophyoid mite interactions: specific and unspecific morphological alterations. Part II. AB - The paper presents recent advances related to both specific and unspecific morphological alterations of plant organs caused by eriophyoid mites. Based on old and new case studies, the diversity of plant malformations, such as galls, non-distortive feeding effects and complex symptoms induced by eriophyoids and/or pathogens vectored by them, is analysed and summarised. PMID- 20012343 TI - Effectiveness of a targeted occupational health intervention in workers with high risk of sickness absence: baseline characteristics and adherence as effect modifying factors in a randomized controlled trial. AB - INTRODUCTION: In a recently published randomized controlled trial (RCT), a targeted occupational health (OH) intervention was found effective in an intention-to-treat analysis in controlling sickness absence among workers with high risk of sickness absence, compared to usual care. We performed an exploratory subgroup analysis in order to detect possible effect modifiers and mediators. METHODS: Age, gender, working status, severity of physical impairment, depression score, self-rated working ability, co-morbidity, and sickness absence characteristics in the previous 12 months were identified as potential effect modifiers (n = 382). We conducted regression analyses with the potential effect modifiers and a mediator (treatment attendance as intended) as explanatory variables. The difference of sickness absence days during the previous year and the follow-up year was the dependent variable. RESULTS: The intervention was especially effective in the subgroups of workers who were certain that they will not be able to continue working in their current job due to health-related reasons (-74 days; 95% CI -105 to -43), had co-morbidities (-22.5 days; 95% CI 35.5 to -9.5), or severe physical impairment at work (-17.5 days; 95% CI -28.5 to -6.5). A modifying effect of age, gender, working status, depressive symptoms, or prior sickness absence on the effectiveness of this OH intervention was not found. CONCLUSIONS: This targeted OH intervention seems especially suitable for workers who consider that they are no longer able to continue working due to health reasons and have high level of physical impairment or co-morbidities. The findings from these exploratory analyses should be tested in future RCTs. PMID- 20012344 TI - Imagining reproduction: the politics of reproduction, technology and the woman machine. AB - Scholars widely assume that the term generation, is preferable to reproduction in the context of early modern history, based on the premise that reproduction to mean procreation was not in use until the end of the eighteenth century. This shift in usage presumably corresponds to the rise of mechanistic philosophy; feminist scholarship, particularly that deriving from the hostile critique fashionable in the 1980s has claimed reproduction is associated with medical practitioners' perceptions of women as baby-producing machines. However, this interpretation, whether in the interests of gender politics or reiterated in more sympathetic histories, misrepresents the historical record. PMID- 20012346 TI - Maternal influences on nausea and vomiting in early pregnancy. AB - Symptoms of nausea and vomiting in early pregnancy (NVP) are common among pregnant women, but whether some women are more likely than others to experience these symptoms has not been well established. We examined potential risk factors for NVP symptom severity, timing of onset, and duration. We included 2,407 newly pregnant women who participated in a prospective cohort study on early pregnancy health between 2000 and 2004 in three U.S. cities. Data on NVP and other health information were collected through telephone interviews, early gestation ultrasound, and medical record abstractions. Generalized linear models were used to model possible risk factors for each NVP characteristic. Eighty-nine percent of women had NVP; for 99% of these, symptoms started in the first trimester. None of the characteristics examined were associated with having NVP. Among those with NVP, increasing risk of delayed symptoms onset was associated with advancing maternal age; increased risks were also seen among non-Hispanic Black [Risk ratio (RR) = 4.3, 95% confidence interval (CI): 1.6,11.6] and Hispanic women (RR = 2.3, 95% CI:0.4,11.5). NVP symptoms for multigravidae were more likely to last beyond the first trimester with each additional pregnancy. Most pregnant women experienced NVP. Nearly all of them, regardless of characteristics examined, had symptoms beginning in the first trimester. Maternal age, race/ethnicity, and gravidity were associated with delayed onset and symptoms that persisted into the second trimester. PMID- 20012345 TI - Examining youth hopelessness as an independent risk correlate for adolescent delinquency and violence. AB - Prior research has identified a relationship between youth hopelessness and violence perpetration within specific groups of young people. The purpose of this study was to evaluate the relationship between youth hopelessness and violence perpetration in a population-based sample of adolescents. This study is a cross sectional analysis of data from 136,549 students in the 6th, 9th, and 12th grades responding to the 2007 Minnesota Student Survey. Logistic regression models were used to evaluate the relationship between hopelessness and youth violence perpetration, including comparison analyses for gender and race/ethnic subgroups as well as adjustment for a measure of poor-low affect. One in four youth (25.1%) reported levels of hopelessness at least enough to bother them in the previous month. Moderate-high levels of hopelessness exhibited a statistically significant independent relationship with a range of violence-related outcomes for youth subgroups: delinquent behavior, weapon carrying on school property, and all forms of self-directed violence. Relationships between hopelessness and interpersonal and intimate partner violence suggest a greater contribution by poor affective functioning for some groups. Interventions designed to reduce youth violence perpetration may benefit from increased strategies to address youth hopelessness as well as youth mental health in general. PMID- 20012347 TI - Adult care transitioning for adolescents with special health care needs: a pivotal role for family centered care. AB - To examine the relationship between having a usual source of care, family centered care, and transition counseling for adolescents with special health care needs. Data are from 18,198 parents/guardians, of youth aged 12-17 years, who participated in the 2005-2006 National Survey of Children With Special Health Care Needs. Linear and logistic regression models were used to define relationships between parent report of identification of a usual place and provider of medical care for their child and counseling on four transition issues: transfer to adult providers, review of future health needs, maintaining health insurance in adulthood, and youth taking responsibility for care. The direct mediating effect of family centered care was evaluated. Youth having a usual source of care (vs. not) were more likely to receive counseling on future health needs (47.4 vs. 33.6%, P < 0.001) and taking responsibility for their own care (79.3 vs. 64.4%, P < 0.001). Having a high level of family centered care (vs. low) was also associated with high rates of discussing future health needs (56.3 vs. 39.6%, P < 0.001) and encouragement to take responsibility for care (91.2 vs. 70.3%, P < 0.001). Family centered care mediated 39.1% of the effect of a usual source of care on discussion of future health needs and 94.9% of the effect of a usual source of care on encouragement to take responsibility for care. Study findings support the development of health care delivery models focusing on family centered care to the same degree as other health care access issues. PMID- 20012348 TI - Intention to become pregnant and low birth weight and preterm birth: a systematic review. AB - Increased stress, psychosocial problems, economic disadvantages, and lack of prenatal care are proposed to explain discrepancies in the outcome of unintended pregnancies. Studies of maternal intention and pregnancy outcomes have yielded varied results. Objective is to review studies of the risk of low birth weight (LBW)/preterm births (PTB) associated with unintended pregnancies ending in a live birth. We reviewed studies reporting on maternal intentions and outcomes from Medline, Embase, CINAHL, and bibliographies of identified articles. An unintended pregnancy was further classified as mistimed (not intended at that time) or unwanted (not desired at any time). Studies reporting an association between pregnancy intention and any of the outcomes were included. Study quality was assessed for biases in selection, exposure assessment, confounder adjustment, analyses, outcomes assessment, and attrition. Unadjusted and adjusted data from included studies were extracted by two reviewers. There were significantly increased odds of LBW among unintended pregnancies [odds ratio (OR) 1.36, 95% confidence interval (CI) 1.25, 1.48] ending in a live birth. Within the unintended category, mistimed (OR 1.31, 95% CI 1.13, 1.52) and unwanted (OR 1.51, 95% CI 1.29, 1.78) pregnancies were associated with LBW. There were statistically significantly increased odds of PTB among unintended (OR 1.31, 95% CI 1.09, 1.58), and unwanted (OR 1.50, 95% CI 1.41, 1.61) but not for mistimed (OR 1.36, 95% CI 0.96, 1.93) pregnancies. Unintended, unwanted, and mistimed pregnancies ending in a live birth are associated with a significantly increased risk of LBW and PTB. PMID- 20012349 TI - Effects of repetitive transcranial magnetic stimulation on spike pattern and topography in patients with focal epilepsy. AB - Repetitive Transcranial Magnetic Stimulation (rTMS) is a non-invasive method for brain stimulation. Group-studies applying rTMS in epilepsy patients aiming to decrease epileptic spike- or seizure-frequency have led to inconsistent results. Here we studied whether therapeutic trains of rTMS have detectable effects on individual spike pattern and/or frequency in patients suffering from focal epilepsy. Five patients with focal epilepsy underwent one session of rTMS online with EEG using a 6 Hz prime/1 Hz rTMS protocol (real and sham). The EEG was recorded continuously throughout the stimulation, and the epileptic spikes recorded immediately before (baseline) and after stimulation (sham and real) were subjected to further analysis. Number of spikes, spike-strength and spike topography were examined. In two of the five patients, real TMS led to significant changes when compared to baseline and sham (decrease in spike-count in one patient, change in topography of the after-discharge in the other patient). Spike-count and topography remained unchanged the remaining patients. Overall, our results do not indicate a consistent effect of rTMS stimulation on interictal spike discharges, but speak in favor of a rather weak and individually variable immediate effect of rTMS on focal epileptic activity. The individuation of most effective stimulation patterns will be decisive for the future role of rTMS in epilepsies and needs to be determined in larger studies. PMID- 20012350 TI - Methodology for combined TMS and EEG. AB - The combination of transcranial magnetic stimulation (TMS) with simultaneous electroencephalography (EEG) provides us the possibility to non-invasively probe the brain's excitability, time-resolved connectivity and instantaneous state. Early attempts to combine TMS and EEG suffered from the huge electromagnetic artifacts seen in EEG as a result of the electric field induced by the stimulus pulses. To deal with this problem, TMS-compatible EEG systems have been developed. However, even with amplifiers that are either immune to or recover quickly from the pulse, great challenges remain. Artifacts may arise from the movement of electrodes, from muscles activated by the pulse, from eye movements, from electrode polarization, or from brain responses evoked by the coil click. With careful precautions, many of these problems can be avoided. The remaining artifacts can be usually reduced by filtering, but control experiments are often needed to make sure that the measured signals actually originate in the brain. Several studies have shown the power of TMS-EEG by giving us valuable information about the excitability or connectivity of the brain. PMID- 20012351 TI - Epithelial cell adhesion molecule (EpCAM) is overexpressed in breast cancer metastases. AB - EpCAM (CD326) has diverse roles in cell adhesion and proliferation, and is known to be overexpressed in primary breast carcinomas (PBCs). While clinical and preclinical data suggest a role for EpCAM in metastases, the only prior study of EpCAM expression in breast cancer metastases suggested that EpCAM expression is decreased after first-line chemotherapy. This study evaluates EpCAM expression in metastatic breast carcinoma (MBC) versus matched PBC . Rapid autopsies were performed on 17 patients with widely metastatic breast cancer. Single patient tissue microarrays (TMAs) were constructed from archived PBC and post-mortem MBCs. In total, 169 spots from 17 PBCs and 895 spots from 195 MBCs were labeled for EpCAM by immunohistochemistry (IHC). Expression was scored as intensity (1-3) multiplied by percent membrane labeling (0-100%) and was subclassified as low (0 100), moderate (101-200), or high (201-300) labeling. PBCs exhibited exclusively low-moderate EpCAM labeling. EpCAM labeling was present in all metastases and was significantly increased in MBCs of 14 of 17 patients (P value range <0.05 to <0.0001, t test). In the remaining three patients, EpCAM labeling was nonsignificantly increased in 1 and unchanged in 2. High EpCAM labeling was verified using a different antibody for IHC, as well as in a separate series of surgically resected metastases compared to unmatched surgically resected primary breast cancers. In conclusion, EpCAM is highly expressed in MBCs compared to matched PBCs, verifying that it is a promising therapeutic target. PMID- 20012353 TI - Inhibition of mammary tumor growth by estrogens: is there a specific role for estrogen receptors alpha and beta? AB - To evaluate the extent to which each estrogen receptor (ER) subtype contributes to the stimulation or to the inhibition of mammary tumor growth, we evaluated the effects of specific agonists in MC4-L2 cells, which are stimulated by 17beta estradiol (E(2)), and in mammary carcinomas of the MPA mouse breast cancer model, which are inhibited by E(2). Both express ERalpha and ERbeta. In MC4-L2 cells, 4,4',4"-(4-propyl-(1H)-pyrazole-1,3,5-triyl)trisphenol (PPT; ERalpha agonist) and (4-hydroxy-phenyl)-propionitrile (DPN; ERbeta agonist) stimulated cell proliferation, whereas the opposite occurred in C4-HI primary cultures. The inhibitory effect was associated with a decrease in ERalpha and cyclin D1 expression and an increase in progesterone receptor (PR) expression as well as in the Bax/Bcl-xl ratio. In vivo, mice carrying C4-HI or 32-2-HI tumors were treated with E(2), PPT or DPN (3 mg/kg/day) or with vehicle. PPT and DPN inhibited tumor size, as did E(2), during the first 72 h. After a few days, DPN-treated tumors started to grow again, while PPT-treated tumors remained quiescent for a longer period of time. A pronounced decrease in the mitotic index and an increase in the apoptotic index was associated with tumor regression. All treated tumors showed: (a) an increase in integrin alpha6 and Bax expression, (b) an increased stromal laminin redistribution, and (c) a decrease in ERalpha, Bcl-xl and Bcl-2 expression (P < 0.001). Apoptosis-inducing factor (Aif) expression was increased in DPN-treated tumors, while active caspase 9 was up-regulated in PPT-treated mice, demonstrating the involvement of the intrinsic apoptotic pathway in estrogen-induced regression in this model. In conclusion, our data indicate that although there may be some preferences for activation pathways by the different agonists, the stimulatory or inhibitory effects triggered by estrogens are cell context dependent rather than ER isoform dependent. PMID- 20012354 TI - A phase II trial of trastuzumab plus weekly ixabepilone and carboplatin in patients with HER2-positive metastatic breast cancer: an Eastern Cooperative Oncology Group Trial. AB - The epothilone B analogue, ixabepilone, binds to b-tubulin, is effective for taxane-refractory metastatic breast cancer (MBC), and may be given every 3 weeks or weekly. We evaluated the efficacy of weekly ixabepilone (I) plus trastuzumab (T) and carboplatin (C) as first line therapy in HER2 + MBC. Patients with HER2+ (3+ by IHC or FISH amplified) MBC received I (15 mg/m2 IV) and C (area under the curve, AUC = 2 IV) on days 1, 8, and 15 of a 28-day cycle for a maximum of 6 cycles, plus weekly T (4 mg/kg loading dose then 2 mg/kg IV) during chemotherapy then every 3 weeks (6 mg/kg IV) until disease progression. The primary objective was to determine whether the combination was associated with a response rate (RR) of at least 75%. Fifty-nine patients were treated, and 39 had HER2 overexpression confirmed in a central lab (cHER2+). For all treated patients, objective response occurred in 26 patients (44%; 95% CI 31-58%), median time to progression was 8.2 months (95% CI 6.3-9.9), and median overall survival was 34.7 months (95% CI 25.7 to [not reached]). Results were comparable for cHer2? cancers. Grade 3-4 adverse events included neutropenia (49%), thrombocytopenia (14%), fatigue (12%), nausea (7%), diarrhea (7%), and neuropathy (7%). One patient died from treatment complications during cycle 1. Weekly ixabepilone and carboplatin plus trastuzumab have an acceptable toxicity profile, but are not likely to be associated with an RR of 75% in HER2+ MBC. Efficacy appears comparable to paclitaxel, carboplatin, and trastuzumab. PMID- 20012356 TI - Survival analysis, longitudinal analysis and causal inference. PMID- 20012355 TI - Gene expression pathway analysis to predict response to neoadjuvant docetaxel and capecitabine for breast cancer. AB - Neoadjuvant chemotherapy has been shown to be equivalent to post-operative treatment for breast cancer, and allows for assessment of chemotherapy response. In a pilot trial of docetaxel (T) and capecitabine (X) neoadjuvant chemotherapy for Stage II/III BC, we assessed correlation between baseline gene expression and tumor response to treatment, and examined changes in gene expression associated with treatment. Patients received four cycles of TX. Tumor tissue obtained from Mammotome core biopsies pretreatment (BL) and post-cycle 1 (C1) of TX was FLash frozen and stored at -70 degrees C until processing. Gene expression analysis utilized Affymetrix HG-U133 Plus 2.0 GeneChip arrays. Statistical analysis was performed using BRB Array Tools after RMA normalization. Gene ontology (GO) pathway analysis used random variance t tests with a significance level of P?0.005. For gene categories identified byGO pathway analysis as significant, expression levels of individual genes within those pathways were compared between classes using univariate t tests; those genes with significance level of P?0.05 were reported. PAM50 analyses were performed on tumor samples to investigate biologic subtype and risk of relapse (ROR). Using GO pathway analysis, 39 gene categories discriminated between responders and non-responders,most notably genes involved in microtubule assembly and regulation. When comparing pre- and post chemotherapy specimens, we identified 71 differentially expressed gene categories, including DNA repair and cell proliferation regulation. There were 45 GO pathways in which the change in expression after one cycle of chemotherapy was significantly different among responders and nonresponders. The majority of tumor samples fell into the basal like and luminal B categories. ROR scores decreased in response to chemotherapy; this change was more evident in samples from patients classified as responders by clinical criteria. GO pathway analysis identified a number of gene categories pertinent to therapeutic response, and may be an informative method for identifying genes important in response to chemotherapy. Larger studies using the methods described here are necessary to fully evaluate gene expression changes in response to chemotherapy. PMID- 20012357 TI - Semiparametric proportional means model for marker data contingent on recurrent event. AB - In many biomedical studies with recurrent events, some markers can only be measured when events happen. For example, medical cost attributed to hospitalization can only incur when patients are hospitalized. Such marker data are contingent on recurrent events. In this paper, we present a proportional means model for modelling the markers using the observed covariates contingent on the recurrent event. We also model the recurrent event via a marginal rate model. Estimating equations are constructed to derive the point estimators for the parameters in the proposed models. The estimators are shown to be asymptotically normal. Simulation studies are conducted to examine the finite-sample properties of the proposed estimators and the proposed method is applied to a data set from the Vitamin A Community Trial. PMID- 20012358 TI - Additive transformation models for clustered failure time data. AB - We propose a class of additive transformation risk models for clustered failure time data. Our models are motivated by the usual additive risk model for independent failure times incorporating a frailty with mean one and constant variability which is a natural generalization of the additive risk model from univariate failure time to multivariate failure time. An estimating equation approach based on the marginal hazards function is proposed. Under the assumption that cluster sizes are completely random, we show the resulting estimators of the regression coefficients are consistent and asymptotically normal. We also provide goodness-of-fit test statistics for choosing the transformation. Simulation studies and real data analysis are conducted to examine the finite-sample performance of our estimators. PMID- 20012359 TI - The effects of N-acetylcysteine on spermatogenesis and degree of testicular germ cell apoptosis in an experimental model of varicocele in rats. AB - INTRODUCTION: The mechanism by which varicocele interferes in spermatogenesis has not been clearly defined. Germ cell apoptosis and oxidative stress appear to be involved in this process and the use of antioxidants has been proposed to counteract upon these effects. The present study evaluated the effects of N acetylcysteine (NAC) on spermatogenesis and germ cell apoptosis in an experimental model of varicocele in rats. MATERIALS AND METHODS: Twenty 30-day old animals were randomly divided into three groups: sham operation (Group 1), left experimental varicocele (Group 2) and left experimental varicocele group treated with NAC 50 mg/kg/day (Group 3). After 2 months, spermatogenesis was evaluated by absolute and true count of round spermatids, pachytenes, spermatocytes and Sertoli cells. The different cell relations were also analyzed. Germ cell apoptosis was quantified using the TUNEL method. The apoptotic index (AI) was calculated as the number of apoptotic cells per tubule. Statistical analysis was performed by analysis of variance considering P < 0.05. RESULTS: The absolute and true cell counts were similar among the groups (P > 0.05). The round spermatid/pachytene ratio was significantly smaller in Groups 2 and 3 compared to the Group 1 (P = 0.012). The AI values were 0.207 +/- 0.09, 0.138 +/- 0.11 and 0.298 +/- 0.27, respectively (P = 0.256). CONCLUSION: Experimental varicocele in rats presented an association with the decreased round spermatid/pachytene ratio, suggesting the loss of germ cells during spermatogenesis. These effects were not influenced by the administration of NAC. Germ cell apoptosis was not influenced by experimental varicocele. PMID- 20012360 TI - An unusual case of extensive epididymotesticular malakoplakia in a diabetic patient. AB - A 67-year-old Saudi man with poorly controlled diabetes-mellitus underwent orchiectomy for a painful left testicular swelling that did not respond to antibiotics. Clinical diagnosis was testicular malignancy. Histologic workup demonstrated extensive involvement of the testis and epididymis by diffuse infiltrate of large histiocytes with eosinophilic granular cytoplasm and numerous Michaelis-Gutmann bodies, which were diagnostic of malakoplakia. Very rarely, malakoplakia has been described in association with diabetes mellitus. Such an extensive malakoplakia occurring at an unusual site in association with systemic illness lends support to the view that a compromised immune status could be etiologically important in malakoplakia. We present this case to highlight the importance of diagnosing malakoplakia when it occurs at unusual locations. We also explore the role of diabetes mellitus in the pathogenesis of malakoplakia and compare the relationship between malakoplakia and xanthogranulomatous inflammation, which are closely related to each-other on both clinical and morphologic grounds. PMID- 20012362 TI - Electronic screening of medical records to detect inpatients at risk of drug related problems. AB - OBJECTIVE: The objective of the study is to develop and validate a screening tool for clinical pharmacists (CPs), comprising electronic queries (EQs) structured to identify inpatients at risk of drug-related problems (DRPs) [1]. SETTING: Internal Medicine and Geriatric Units of the (multi-site) Hopital du Valais. METHOD: EQs, identifying patients with potential DRPs and/or those requiring a medication review were compared with a CP-conducted manual check of all electronic medical records (EMRs), during their standard activities. The queries were aimed at identifying patients receiving drugs such as cytochrome P450 inducers, inhibitors or high-risk medications, those with renal impairment, those on digoxin with low serum potassium, those with intravenous (i.v.) anti infectives or i.v. acetaminophen for more than 3 days, and elderly patients with polymedication (>or=80 years and >10 drugs). MAIN OUTCOME MEASURE: Sensitivity and specificity of the screening tool. RESULTS: Five hundred and one patients seen by 4 CPs during 36 ward rounds were screened. The EQs identified 64.7% of the visited inpatients. The CPs detected at least one DRP in 64.9% of all patients. A sensitivity of 85.1% and a specificity of 60.4% were achieved. CONCLUSION: This tool allows for efficient and rapid screening of patients at risk of DRPs in preparation for the ward round. It helps CPs to prioritize their medication reviews and to optimize their workload. PMID- 20012363 TI - Clinical pharmacy services and solid organ transplantation: a literature review. AB - AIM OF THE REVIEW: Organ transplantation represents the therapy of choice for most types of end-stage organ failure, and post-transplant patient care warrants great attention. The aim of this study was to summarise the available evidence regarding the role and impact of clinical pharmacy services in the care of solid organ transplant patients. METHODS: A search of the literature was conducted using the MEDLINE, EMBASE and IPA databases to identify studies relevant to our investigation of the impact of clinical pharmacists' interventions. RESULTS: Only five out of nineteen of the included studies were randomised controlled trials; eleven studies were descriptive, and three were before-after studies. Interventions performed in these studies consisted of routine clinical pharmacy services with a focus on identifying, resolving and preventing drug-related problems; clinical pharmacy services with a focus on therapeutic drug monitoring; and those with a focus on compliance enhancement and educational interventions. The number and type of interventions and the physicians' acceptance rates were assessed in the majority of the included studies. Acceptance rates were generally above 95%, and most studies reported that clinical pharmacy services had a positive impact on the care of solid organ transplant patients. Positive perceptions of patients and health care professionals are also reported. In two of the studies, patients' compliance rates and drug knowledge were assessed following counselling by a pharmacist. Dosing-related interventions were the most common interventions proposed. Immunosuppressants, cardiovascular drugs and antimicrobials were the drug classes most affected by the clinical pharmacists' interventions. CONCLUSIONS: High quality evidence that supports the benefit of clinical pharmacy services in the care of solid organ transplant patients is rare. Nevertheless, all of the included studies showed that clinical pharmacy services had a positive impact. Furthermore, all included studies showed that patients and physicians appreciated clinical pharmacists. The various outcome measures used in these studies were improved by interactions with clinical pharmacists. More randomised controlled trials are needed to contribute to the paucity of the existing evidence. PMID- 20012364 TI - Host cell death induced by the egress of intracellular Plasmodium parasites. AB - Intracellular pathogens are known to inhibit host cell apoptosis efficiently to ensure their own survival. However, following replication within a cell, they typically need to egress in order to infect new cells. For a long time it was assumed that this happens by simply disrupting the host cell and in some cases, such as for Plasmodium-infected erythrocytes, this seems indeed to be true. However, recently it has been shown that in Plasmodium-infected hepatocytes, an ordered form of cell death is initiated. This cell death is parasite-dependent and can clearly be distinguished from apoptosis and necrosis. The key event, and point of no return, appears to be the rupture of the parasitophorous vacuole membrane (PVM). PVM disruption and host cell death depend on the activation of cysteine proteases. Whether these are of parasite or host cell origin seems to rely on the life cycle stage of the Plasmodium parasite and the corresponding host cell. PMID- 20012365 TI - Elucidating the role of requiem in the growth and death of Chinese hamster ovary cells. AB - Requiem, a hypothesized transcription factor with apoptosis-related activity, was previously shown to be a potential cell engineering gene target for improving recombinant protein production. Requiem suppression has resulted in improved viable cell density and extended culture viability, leading to an overall improvement in recombinant protein productivity. However, not much is known about the function of requiem. We found that requiem is highly conserved at both nucleotide and amino acid levels in Chinese hamster ovary (CHO) cells when compared to human and mouse sequences, suggesting that requiem's functional role is evolutionary well conserved. Upon inducing requiem over-expression, proliferation rates of CHO cells were significantly decreased with doubling times increased by 26%. Interestingly, the over-expression of requiem did not decrease cell viability and could not induce apoptosis. However, requiem sensitized the cells to increased caspase-9 activities under staurosporine-induced apoptosis, suggesting that it has a role to play in mitochondria-mediated apoptosis under staurosporine treatment. The nuclear localization of REQUIEM in CHO cells and its conserved plant homeodomain (PHD) zinc fingers seem to further support the hypothesis that requiem encodes for a potential transcription factor. Upon requiem over-expression, we found that the differentially expressed genes involved in transcriptional regulation and cell proliferation and growth were associated both upstream and downstream of p53. PMID- 20012361 TI - Posttraumatic stress disorder in maltreated youth: a review of contemporary research and thought. AB - Youths who have been maltreated often experience symptoms of posttraumatic stress disorder (PTSD), and this special population has received increased attention from researchers. Pathways toward maladaptive effects of maltreatment and PTSD are remarkably similar and reflect specific biological diatheses and psychological vulnerabilities that produce wide-ranging self-regulation deficits. Developmental models of effects of maltreatment and of PTSD are thus increasingly intertwined and have begun to inform specialized assessment and treatment strategies for this population. This review covers key aspects of posttraumatic stress disorder in maltreated youth, including epidemiology, symptomatology, outcome, and risk factors as well as assessment and treatment strategies and challenges for these youths. PMID- 20012366 TI - Molecular analysis of Candida albicans isolates from clinical specimens. AB - The aim of this study was to genotype Candida albicans strains isolated from various clinical specimens by using CA-INT-R and CA-INT-L primer pairs designed to span the region that includes the site of the transposable group-1 intron in the 25S rRNA gene. A total of 194 C. albicans isolates (28 invasive and 166 noninvasive) were genotyped. The frequencies of genotypes A, B, C and D were found as 51.0, 29.4, 19.1 and 0.5%, respectively. Statistically significant difference was determined between frequency of genotype distribution between invasive and noninvasive isolates (P < 0.001). Genotype C was more prevalent among invasive isolates while genotype A was in noninvasive ones. Furthermore, six different subtypes were determined among genotype A C. albicans isolates by restriction endonuclease analysis using a previously constructed differentiation scheme consisting of HaeIII and MspI digestions. This study demonstrated the genetic diversity of clinical isolates of C. albicans in our hospital. PMID- 20012367 TI - Presence of C. albidus, C. laurentii and C. uniguttulatus in crop and droppings of pigeon lofts (Columba livia). AB - Columba livia is an important reservoir and carrier of Cryptococcus neoformans, Cryptococcus uniguttulatus, Cryptococcus laurentii and Cryptococcus albidus. Upper digestive tract of this species is also known as a habitat for Cryptococcus neoformans. Given the increasing clinical interest of this microorganism, 331 swabs from crop and 174 dropping samples from pigeon lofts in Grand Canary Island have been studied. The obtained results show an extensive presence samples 81 positive (24.47%) of Cryptococcus spp. in analysed crops: 32 (9.66%) for C. neoformans, 24 (7.2%) for C. uniguttulatus, 23 (6.9%) for C. albidus and 2 (0.6%) for C. laurentii. In the same way, Cryptococcus spp was also isolated in 82 (47.13%), dropping samples: C. neoformans in 59 (33.9%), C. uniguttulatus, in 9 (5.17%), C. laurentii in 8 (4.59%) and C. albidus in 6 (3.44%) of the investigated samples, respectively. The cryptococcosis produced by species of cryptococci other than C. neoformans has become more important during the last decade, supporting the study on the role of pigeon in the epidemiology of this disease. PMID- 20012368 TI - The complete mitochondrial genome of the cockroach Eupolyphaga sinensis (Blattaria: Polyphagidae) and the phylogenetic relationships within the Dictyoptera. AB - We present the complete mitochondrial DNA sequence of Eupolyphaga sinensis. This closed circular molecule is 15553 bp long and consists of 37 genes that encode for 13 inner membrane proteins, 2 ribosomal RNAs and 22 transfer RNAs. The genome shares the gene order and orientation with previously known Blattaria mitochondrial genomes. All tRNAs could be folded into the typical cloverleaf secondary structure, but the tRNASer (AGN) appears to be missing the DHU arm. The A + T-rich region is 857 bp long and longer than other cockroaches. Based on the concatenated amino acid sequences of all protein coding genes of E. sinensis in conjunction with those 23 other arthropod sequences, we reconstruct the phylogenetic tree. Phylogenetic analyses shows that Blataria (including Isoptera) and the Mantodea are sister groups. Furthermore the relationship of the three basal clades of winged insects are different from the three previous hypotheses ((Ephemeroptera + Odonata) +Neoptera, Ephemeroptera + (Odonata + Neoptera), Odonata + (Ephemeroptera +Neoptera)). The Ephemeroptera (Parafronurus youi) clusters with the Plecoptera (Pteronarcys princes). PMID- 20012369 TI - Alternative splicing and gene polymorphism of the human TAP3/SEC14L4 gene. AB - Three closely related human SEC14p-like proteins (hTAP1, hTAP2, hTAP3, or SEC14L2, SEC14L3, SEC14L4, respectively) have been described that are related to the Saccharomyces cerevisiae SEC14 protein. These proteins may participate in intracellular lipid transport and influence regulatory lipid-dependent events. Here we report the isolation of an alternatively spliced hTAP3 cDNA and a polymorphism within the coding region of the hTAP3/SEC14L4 gene. PMID- 20012370 TI - Molecular cloning and functional expression analysis of a new gene encoding geranylgeranyl diphosphate synthase from hazel (Corylus avellana L. Gasaway). AB - Geranylgeranyl diphosphate synthase (GGPPS) [EC 2.5.1.29] catalyzes the biosynthesis of geranylgeranyl diphosphate (GGPP), which is a key precursor for diterpenes such as taxol. Herein, a full-length cDNA encoding GGPPS (designated as CgGGPPS) was cloned and characterized from hazel (Corylus avellana L. Gasaway), a taxol-producing angiosperms. The full-length cDNA of CgGGPPS was 1515 bp with a 1122 bp open reading frame (ORF) encoding a 373 amino acid polypeptide. The CgGGPPS genomic DNA sequence was also obtained, revealing CgGGPPS gene was not interrupted by an intron. Southern blot analysis indicated that CgGGPPS belonged to a small gene family. Tissue expression pattern analysis indicated that CgGGPPS expressed the highest in leaves. RT-PCR analysis indicated that CgGGPPS expression could be induced by exogenous methyl jasmonate acid. Furthermore, carotenoid accumulation was observed in Escherichia coli carrying pACCAR25DeltacrtE plasmid carrying CgGGPPS. The result revealed that cDNA encoded a functional GGPP synthase. PMID- 20012371 TI - Delisheng, a Chinese medicinal compound, exerts anti-proliferative and pro apoptotic effects on HepG2 cells through extrinsic and intrinsic pathways. AB - The anti-proliferative, cytotoxic and apoptogenic activities of delisheng, a Chinese medicinal compound, has been investigated. In this study, the hepatocarcinoma cell line (HepG2) and the liver cell line (L-02) were exposed to delisheng (6.25, 50 and 100 MUl/ml). Delisheng suppressed the proliferation and viability of normal liver L-02 cells slightly, but strongly inhibited the proliferation and viability of hepatocarcinoma HepG2 cells. The flow cytometric analysis of HepG2 cells demonstrated that delisheng primarily arrested the HepG2 cells at the G1 phase of the cell cycle. Annexin V-FITC/PI staining corroborates the apoptogenic nature of delisheng on HepG2 cells. The anti-proliferative and pro-apoptotic effect of delisheng in HepG2 cells was associated with changes in the Bcl-2/Bax ratio and the induction of caspase-mediated apoptosis. Upregulation of DR5 expression was observed in HepG2 cells after treatment with delisheng. The findings from the present study suggest that delisheng has selective cytotoxic activities against HepG2 cells. Delisheng triggered time- and dose-dependent apoptosis in HepG2 cells by activating the mitochondria-mediated and death receptor-mediated apoptotic pathways. PMID- 20012373 TI - Upregulation of cytosolic NADP+-dependent isocitrate dehydrogenase by hyperglycemia protects renal cells against oxidative stress. AB - Hyperglycemia-induced oxidative stress is widely recognized as a key mediator in the pathogenesis of diabetic nephropathy, a complication of diabetes. We found that both expression and enzymatic activity of cytosolic NADP(+)-dependent isocitrate dehydrogenase (IDPc) were upregulated in the renal cortexes of diabetic rats and mice. Similarly, IDPc was induced in murine renal proximal tubular OK cells by high hyperglycemia, while it was abrogated by co-treatment with the antioxidant N-Acetyl-Cysteine (NAC). In OK cells, increased expression of IDPc by stable transfection prevented hyperglycemia-mediated reactive oxygen species (ROS) production, subsequent cellular oxidative stress and extracellular matrix accumulation, whereas these processes were all stimulated by decreased IDPc expression. In addition, production of NADPH and GSH in the cytosol was positively correlated with the expression level of IDPc in OK cells. These results together indicate that upregulation of IDPc in response to hyperglycemia might play an essential role in preventing the progression of diabetic nephropathy, which is accompanied by ROS-induced cellular damage and fibrosis, by providing NADPH, the reducing equivalent needed for recycling reduced glutathione and low molecular weight antioxidant thiol proteins. PMID- 20012372 TI - Mismatch repair protein expression and colorectal cancer in Hispanics from Puerto Rico. AB - Colorectal cancer (CRC) is a leading cause of morbidity and mortality and alterations in mismatch repair (MMR) genes, leading to absent protein (negative) expression, are responsible for approximately 20% of CRC cases. Immunohistochemistry is a tool for prescreening of MMR protein expression in CRC but the literature on its use on Hispanics is scarce. However, Hispanics represent the second leading ethnicity in the United States (US) and CRC is a public health burden in this group. Our objectives were to determine the frequency of MMR protein-negative CRC and to evaluate its association with clinical and pathological characteristics among Hispanics from Puerto Rico, for the first time to our knowledge. A retrospective observational study of unselected CRC patients from the Puerto Rico Medical Center from 2001 to 2005 was done. MLH1 and MSH2, the most commonly altered MMR genes, protein expression was evaluated using immunohistochemistry, with microsatellite instability (MSI) and BRAF gene analyses in the absence of MLH1 protein expression. One-hundred sixty four CRC patients were evaluated: the overall MMR protein-negative frequency was 4.3%, with 0.6% frequency of co-occurrence of MLH1-protein negative expression, MSI-high, and normal BRAF gene. MMR protein-negative expression was associated with proximal colon location (P = 0.02) and poor histological tumor differentiation (P = 0.001), but not with other characteristics. The frequency of MMR protein-negative CRC in Hispanics from Puerto Rico was lower than reported in other populations. This finding may explain the lower CRC incidence rate among US Hispanics as compared to US non-Hispanic whites and blacks. PMID- 20012374 TI - Design and ex vivo kidney evaluation of a high-intensity focused ultrasound transducer and 3D positioner. AB - This article describes the design, construction, and evaluation of a three dimensional (3D) ultrasound system to be used for the treatment of kidney tumors using high-intensity focused ultrasound (HIFU). The system consists of a therapeutic ultrasound delivery module (driver, power amplifier, matching circuit, and transducer (1 MHz)) and a 3D positioning module (stepper motors and microcontroller). The system includes software that designs a treatment planning according to the tumor coordinates. In order to verify the capability of the system for moving the transducer, the positioning module was successfully driven to cover cylinders of dimensions as large as 20 cm (diameter) x 10 cm (height) with step sizes as low as 0.05 mm. Several exposimetry experiments were done which showed close match between experimental and theoretical results. Ex vivo experiments were performed and indicated the capability of the system to generate a single lesion of 0.27 cm diameter and 0.45 cm length, as well as five lesions (each with a diameter of 0.3 cm) at prespecified locations. During the ex vivo experiments, temperature monitoring indicated an increase in temperature from 37 degrees C to about 60 degrees C inside the lesion, while outside it the temperature remained below 39 degrees C. PMID- 20012375 TI - Dose-guided radiotherapy for lung tumors. AB - The transit in vivo dosimetry performed by an electronic portal imaging device (EPID) is a very practical method to check error sources in radiotherapy. Recently, the present authors have developed an in vivo dosimetry method based on correlation functions, F (w, L), defined as the ratio between the transit signal, S(t) (w, L), by the EPID and the mid-plane dose, D(m) (w, L), in a solid water phantom as a function of the phantom thickness, w, and of the field dimensions, L. In particular, generalized correlation functions F (w, L) for 6, 10 and 15 MV X-ray beams supplied by a pilot Varian linac, are here used by other three linacs operating in two centers. This way the workload, due to measurements in solid water phantom, needed to implement the in vivo dosimetry method was avoided. This article reports a feasibility study on the potentiality of this procedure for the adaptive radiotherapy of lung tumors treated by 3D conformal radiotherapy techniques. In particular, the dose reconstruction at the isocenter point D(iso) in the lung tumor has been used as dose-guided radiotherapy (DGRT), to detect the inter-fraction tumor anatomy variations that can require new CT scans and an adaptive plan. When a difference greater than 6% between the predicted dose by the treatment planning system (TPS), D (iso,TPS) and the D(iso) was observed, the clinical action started to detect possible anatomical lung tumor changes. Twelve over twenty patients examined presented in vivo dose discrepancies due to the tumor morphological changes during treatments, and these results were successively confirmed by new CT scans. In this work, for a patient that showed for all beams, D (iso) values over the tolerance level, the new CT scan was used for an adaptive plan. The lung dose volume histogram for D (iso,TPS) = 2 Gy per fraction suggested the adaptive plan. In particular, the lung volume included in 2 Gy increased from 350 cm(3) of the original plan to 550 cm(3) of the hybrid plan, while for the adaptive plan the lung volume included in 2 Gy decreased to 15 cm(3). Moreover, the mean doses to the organs at risk were reduced to 70%. The results of this research show that the DGRT procedure by the D(iso) reconstruction, integrated with radiological imaging, was feasible for periodic investigation on morphological lung tumor changes. This feasibility study takes into account the accuracy of two algorithms based on the pencil beam and collapsed cone convolution models for dose calculations where large density inhomogeneities are present. PMID- 20012376 TI - Computer-aided assessment of scoliosis on posteroanterior radiographs. AB - In order to reduce the observer variability in radiographic scoliosis assessment, a computer-aided system was developed. The system semi-automatically measured the Cobb angle and vertebral rotation on posteroanterior radiographs based on Hough transform and snake model, respectively. Both algorithms were integrated with the shape priors to improve the performance. The system was tested twice by each of three observers. The intraobserver and interobserver reliability analyses resulted in the intraclass correlation coefficients higher than 0.9 and 0.8 for Cobb measurement on 70 radiographs and rotation measurement on 156 vertebrae, respectively. Both the Cobb and rotation measurements resulted in the average intraobserver and interobserver errors less than 2 degrees and 3 degrees , respectively. There were no significant differences in the measurement variability between groups of curve location, curve magnitude, observer experience, and vertebra location. Compared with the documented results, measurement variability is reduced by using the developed system. This system can help orthopedic surgeons assess scoliosis more reliably. PMID- 20012379 TI - Delay in diagnostic workup and treatment of esophageal cancer. AB - INTRODUCTION: Esophageal cancer should preferably be detected and treated at an early stage, but this may be prohibited by late onset of symptoms and delays in referral, diagnostic workup, and treatment. The aim of this study was to investigate the impact of these delays on outcome in patients with esophageal cancer. METHODS: For 491 patients undergoing esophagectomy for cancer between 1991 and 2007, patients' short- and long-term outcome were analyzed according to different time intervals between onset of symptoms, diagnosis, and surgical treatment. RESULTS: Length of prehospital delay (from onset of symptoms until endoscopic diagnosis) did not affect patient's short- or long-term outcome. A shorter hospital delay between establishing the diagnosis of esophageal cancer on endoscopy and surgery was associated with lower overall morbidity and in-hospital mortality. Patients of ASA classes I and II experienced a shorter hospital delay than patients of ASA classes III and IV. Length of hospital delay between endoscopic diagnosis and surgery did not affect pathological tumor-node metastasis stage or R0-resection rate. Longer hospital delay did not result in worse survival: Overall survival after esophagectomy for cancer was not significantly different between patients with hospital delay <5, 5-8, or >8 weeks (24.7%, 21.7%, and 32.3%, respectively; p = 0.12). CONCLUSION: A longer hospital delay (between endoscopic diagnosis and surgery) resulted in worse patient's short-term outcome (higher overall morbidity and mortality rates) but not in a worse long-term outcome (overall survival). This may be explained by a more time consuming diagnostic workup in patients with a poorer physical status and not by tumor progression. PMID- 20012380 TI - Is there a role for anything other than a Nissen's operation? AB - BACKGROUND: The Nissen fundoplication is the most frequently applied antireflux operation worldwide. The aim of this review was to compare laparoscopic Nissen with partial fundoplication. METHODS: Nine randomized trials comparing several types of wraps were analyzed, four for the comparison Nissen vs. Toupet and five for the comparison Toupet or Nissen vs. anterior fundoplication. Similar comparisons in nonrandomized studies were also included. RESULTS: Dysphagia rates and reflux recurrence were not related to preoperative esophageal persistalsis independent of the selected procedure. Overall, Nissen fundoplication revealed slightly better reflux control, but was associated with more side effects, such as early dysphagia and gas bloat. Advantages of an anterior approach were only reported by one group. A significantly higher reflux recurrence rate for anterior fundoplication was observed in all other comparisons. CONCLUSION: Tailoring antireflux surgery according to esophageal motility is not indicated. At present, the relevant factor for selection of a Nissen or Toupet fundoplication is personal experience. Anterior fundoplication offers less effective long-term reflux control. PMID- 20012381 TI - Responses of wheat roots to exogenous selenium supply under enhanced ultraviolet B. AB - Effects of selenium (Se) on growth and some physiological traits of roots in wheat (Triticum aestivum L. cv Han NO.7086) seedlings exposed to enhanced ultraviolet-B (UV-B) stress are reported. Responses of roots were different depending on the Se concentration. Compared with the control, root weight of wheat seedlings treated with 1.0 and 2.0 mg Se kg(-1) soil increased by 39.47% and 16.28%, respectively. The lower amount Se (0.5 mg kg(-1)) and the higher amount Se treatments (3.0 mg kg(-1)) did not significantly affect on root weight. Se treatments significantly increased root activity, flavonoids and proline content, and activities of peroxidase and superoxide dismutase in wheat roots exposed to enhanced UV-B. In addition, the treatments with 0.5, 1.0, and 2.0 mg Se kg(-1) significantly reduced malondialdehyde content and the rate of superoxide radical (O2(-) ) production of roots, whereas the higher amount Se treatment only induced a decrease in the rate of O2(-) production. The results of this study demonstrated that optimal Se supply promoted roots growth of wheat seedlings, and that optimal Se supply could reduce oxidative stress in wheat roots under enhanced UV-B radiation. PMID- 20012382 TI - Aortic elasticity is impaired in patients with endemic fluorosis. AB - Sixty-three patients with endemic fluorosis (36 males/27 females; mean age 33.9 +/- 8.6 years) and 45 age-, sex-, and body mass index-matched healthy controls (30 males/15 females; mean age 32.7 +/- 8.8 years) were included in this study. Aortic stiffness indices, aortic strain (AS), aortic distensibility (AD), and aortic strain index (ASI) were calculated from the aortic diameters measured by echocardiography and blood pressure obtained by sphygmomanometry. The urine fluoride levels of fluorosis patients were significantly higher than control subjects as expected (1.9 +/- 0.1 mg/l vs. 0.4 +/- 0.1 mg/l, respectively; P < 0.001). AS and AD were significantly lower in fluorosis patients than in the controls (for AS 5.3 +/- 3.6 vs. 8.0 +/- 3.4%; P < 0.001 and for AD 0.2 +/- 0.1 vs. 0.3 +/- 0.1 cm(2) dyn(-1) 10(-3); P < 0.001, respectively). In contrast, significantly higher ASI was observed in fluorosis patients than in the controls (3.4 +/- 0.6 vs. 3.0 +/- 0.4; P < 0.001, respectively). The results of our study demonstrate that elastic properties of ascending aorta are impaired in patients with endemic fluorosis. PMID- 20012383 TI - Cadmium toxicity in spermatogenesis and protective effects of L-carnitine in adult male rats. AB - In this study, the effects of cadmium toxicity and the protective effects of L carnitine on spermatogenesis in Sprague-Dawley rat were evaluated. Animals were subdivided into five groups. Cadmium chloride (1-mg/kg body weight) was injected intraperitoneally during 16 days at intervals of 48 h between subsequent treatments. L-carnitine (500 mg/kg b.w., IP) was pretreated in both of control and cadmium-injected rats. Animals were killed on day 17 after the first treatment. The left cauda epididymis was removed and immediately immersed into Hank's balanced salt solution for evaluation of sperm count and viability. Following contamination with cadmium, a decrease in the number and viability of cauda epididymis sperm, the number of cell proliferation, and Johnsen Scores in the seminiferous tubules was observed. Consequently, L-carnitine treatment caused an increase in the number and viability of cauda epididymis sperm, the number of cell proliferation, and Johnsen Scores in the cadmium-induced group. PMID- 20012384 TI - Salivary fluoride concentrations and fluoride ingestion following application of preparations containing high concentration of fluoride. AB - The aim of the study was to assess salivary fluoride concentrations and ingestion of fluorides after five different techniques of topical fluoride treatment. Ten volunteers received applications of fluoride gel, toothpaste, and foam. Fluoride concentrations were determined using ion-selective fluoride electrode in the samples of unstimulated saliva before and after procedures. The amounts of fluoride applied, recovered from the mouth, and retained in the mouth were calculated for each treatment. It was proved that fluoride ingestion following tray application of fluoride foam was significantly lower than ingestion following tray applications of fluoride gel (p < 0.01). The use of limited amounts of fluorides on a tray resulted in similar fluoride retention and lower ingestion comparing to the method which involves a large portion of fluorides followed by rinsing. Tooth brushing resulted in high salivary retention rates per amount of fluorides used in the procedure. These data provide initial concept about the possible advantages of some methods of topical fluoride application over others. PMID- 20012385 TI - From manganism to manganese-induced parkinsonism: a conceptual model based on the evolution of exposure. AB - Manganism is a distinct medical condition from Parkinson's disease. Manganese exposure scenarios in the last century generally have changed from the acute, high-level exposure conditions responsible for the occurrence of manganism to chronic exposure to much lower levels. Such chronic exposures may progressively extend the site of manganese deposition and toxicity from the globus pallidus to the entire area of the basal ganglia, including the substantia nigra pars compacta involved in Parkinson's disease. The mechanisms of manganese neurotoxicity from chronic exposure to very low levels are not well understood, but promising information is based on the concept of susceptibility that may place individuals exposed to manganese at a higher risk for developing Parkinsonian disturbances. These conditions include mutations of genes which play important pathogenetic roles in both Parkinsonism and in the regulation of manganese transport and metabolism. Liver function is also important in manganese related neurotoxicity and sub-clinical impairment may increase the risk of Parkinsonism. The purpose and scope of this report are to explore the literature concerning manganese exposure and potential subclinical effects and biological pathways, impairment, and development of diseases such as Parkinsonism and manganism. Inhalation and ingestion of manganese will be the focus of this report. PMID- 20012387 TI - HLA complex-linked heat shock protein genes and childhood acute lymphoblastic leukemia susceptibility. AB - Three heat shock protein 70 (HSP70) genes, HSPA1L, HSPA1A, and HSPA1B, are located within the human leukocyte antigen (HLA) class III region. HSPs act as stress signals and regulate natural killer cell response to cancer. HSP70 gene polymorphisms show disease associations partly due to their linkage disequilibrium with HLA alleles. To systematically evaluate their associations with childhood acute lymphoblastic leukemia (ALL), we examined the three functional single nucleotide polymorphisms (SNPs) rs2227956 (T493M) in HSPA1L, rs1043618 in HSPA1A 5'UTR, and rs1061581 (Q351Q) in HSPA1B by TaqMan assays or polymerase chain reaction-restriction fragment length polymorphism in 114 ALL cases and 414 controls from Wales (UK), in 100 Mexican Mestizo ALL cases and 253 controls belonging to the same ethnic group, and in a panel of 82 HLA-typed reference cell line samples. Homozygosity for HSPA1B rs1061581 minor allele G was associated with protection (odds ratio (OR) = 0.37, 95% confidence interval (CI) = 0.16-0.78; P = 0.007) with gene-dosage effect (additive model) reaching significance (P = 0.0001) in the Welsh case-control group. This association was replicated in the second case-control group from Mexico (OR (recessive model) = 0.49, 95% CI = 0.24-0.96; P = 0.03), and the pooled analysis yielded a strong association (Mantel-Haenszel OR = 0.43, 95% CI = 0.27-0.69, P = 0.0004). The association was stronger in males in each group and in the pooled analysis. A three-SNP haplotype including the major allele A of rs1061581 showed a highly significant increase in Welsh cases compared with respective controls (6.7% vs 1.8%; P = 0.0003) due to the difference between male cases and controls. The protective allele of rs1061581 occurred more frequently on the HLA-DRB3 haplotypes (especially DRB1*03) in the cell line panel, but the HSPA1B association was independent from the HLA-DRB4 association previously detected in the same case-control group from Wales (adjusted P = 0.001). Given the cancer promoting roles played by HSPs intracellularly as well as roles in immune surveillance when expressed on the cell surface and the known correlations between expression levels and the HSP polymorphisms, these results are likely to indicate a primary association and warrant detailed assessment in childhood ALL development. PMID- 20012389 TI - Improving toxicity screening and drug development by using genetically defined strains. AB - According to the US Food and Drugs Administration (Food and Drug Administration (2004) Challenge and opportunity on the critical path to new medical products.) "The inability to better assess and predict product safety leads to failures during clinical development and, occasionally, after marketing". This increases the cost of new drugs as clinical trials are even more expensive than pre clinical testing.One relatively easy way of improving toxicity testing is to improve the design of animal experiments. A fundamental principle when designing an experiment is to control all variables except the one of interest: the treatment. Toxicologist and pharmacologists have widely ignored this principle by using genetically heterogeneous "outbred" rats and mice, increasing the chance of false-negative results. By using isogenic (inbred or F1 hybrid, see Note 1) rats and mice instead of outbred stocks the signal/noise ratio and the power of the experiments can be increased at little extra cost whilst using no more animals. Moreover, the power of the experiment can be further increased by using more than one strain, as this reduces the chance of selecting one which is resistant to the test chemical. This can also be done without increasing the total number of animals by using a factorial experimental design, e.g. if the ten outbred animals per treatment group in a 28-day toxicity test were replaced by two animals of each of five strains (still ten animals per treatment group) selected to be as genetically diverse as possible, this would increase the signal/noise ratio and power of the experiment. This would allow safety to be assessed using the most sensitive strain.Toxicologists should also consider making more use of the mouse instead of the rat. They are less costly to maintain, use less test substance, there are many inbred and genetically modified strains, and it is easier to identify gene loci controlling variation in response to xenobiotics in this species.We demonstrate here the advantage of using several inbred strains in two parallel studies of the haematological response to chloramphenicol at six dose levels with CD-1 outbred, or using four inbred strains of mice. Toxicity to the white blood cell lineage was easily detected using the inbred strains but not using the outbred stock, clearly showing the advantage of using the multi-inbred strain approach. PMID- 20012388 TI - Computation of egomotion in the macaque cerebellar vermis. AB - The nodulus and uvula (lobules X and IX of the vermis) receive mossy fibers from both vestibular afferents and vestibular nuclei neurons and are thought to play a role in spatial orientation. Their properties relate to a sensory ambiguity of the vestibular periphery: otolith afferents respond identically to translational (inertial) accelerations and changes in orientation relative to gravity. Based on theoretical and behavioral evidence, this sensory ambiguity is resolved using rotational cues from the semicircular canals. Recordings from the cerebellar cortex have identified a neural correlate of the brain's ability to resolve this ambiguity in the simple spike activities of nodulus/uvula Purkinje cells. This computation, which likely involves the cerebellar circuitry and its reciprocal connections with the vestibular nuclei, results from a remarkable convergence of spatially- and temporally-aligned otolith-driven and semicircular canal-driven signals. Such convergence requires a spatio-temporal transformation of head centered canal-driven signals into an estimate of head reorientation relative to gravity. This signal must then be subtracted from the otolith-driven estimate of net acceleration to compute inertial motion. At present, Purkinje cells in the nodulus/uvula appear to encode the output of this computation. However, how the required spatio-temporal matching takes place within the cerebellar circuitry and what role complex spikes play in spatial orientation and disorientation remains unknown. In addition, the role of visual cues in driving and/or modifying simple and complex spike activity, a process potentially critical for long-term adaptation, constitutes another important direction for future studies. PMID- 20012390 TI - The sophisticated mouse: protecting a precious reagent. AB - Definable, genetically and environmentally, the humble mouse has become a reagent with which to probe the human condition. The information thus gained is leading to a greater understanding of inter-individual variation in drug responses and disease processes and is forming the basis for personalized medicine. Inbred mice are the tool of choice as each strain is essentially clonal in nature creating a defined, uniform setting where the effects of genetic background and modifications can be evaluated coherently. However, the creation and characterization of novel mouse strains remain expensive and time consuming. Further, the continual maintenance of these valuable animals as live colonies is financially draining and carries continual potential risks, including disastrous loss due to fire, flood, disease, etc. There are also other more insidious disasters including genetic contamination and genetic drift, either of which can go undiscovered until their effects ruin experiments. With this in mind, we strongly recommend that all mouse strains be cryopreserved as a matter of standard mouse management. Cryopreservation is a powerful colony management tool, assuring strains are available upon demand, for example, for regulatory requirements, re-initiation of projects, collaborations, re-evaluation of data etc. However, it is essential that any cryopreservation approach be cost effective for both strain closure and strain recovery. In this chapter, we describe the variables which can afflict an inbred mouse's genetic background (and hence phenotype), options to consider for strain archiving, and describe how to economically store and recover strains by sperm cryopreservation. PMID- 20012391 TI - Genetically engineered mouse models in drug discovery research. AB - Genetically modified mouse models have been proven to be a powerful tool in drug discovery. The ability to genetically modify the mouse genome by removing or replacing a specific gene has enhanced our ability to identify and validate target genes of interest. In addition, many human diseases can be mimicked in the mouse and signaling pathways have been shown to be conserved. In spite of these advantages the technology has limitations. In transgenic animals there may be significant heterogeneity among different founders. In knock-out animals the predicted phenotypes are not always readily observed and occasionally a completely novel and unexpected phenotype emerges. To address the latter and ensure that a deep knowledge of the target of interest is obtained, we have developed a comprehensive phenotyping program which has identified novel phenotypes as well as any potential safety concerns which may be associated with a particular target. Finally we continue to explore innovative technologies as they become available such as RNAi for temporal and spatial gene knock-down and humanized models that may better simulate human disease states. PMID- 20012392 TI - Engineering the mouse genome to model human disease for drug discovery. AB - Genetically engineered mice (GEM) have become invaluable tools for human disease modeling and drug development. Completion of the mouse genome sequence in combination with transgenesis and gene targeting in embryonal stem cells have opened up unprecedented opportunities. Advanced technologies for derivation of GEM models will be introduced and discussed. PMID- 20012393 TI - Profiling of drug action using reporter mice and molecular imaging. AB - Reporter mice associated to molecular imaging represent a major asset for the study of the spatio-temporal effects of drugs in living animals. The field is still relatively young and so far the number of animals genetically modified to express a given reporter gene ubiquitously and under the control of specific drugs is still limited. For a reporter animal the indispensable elements for the application to drug research and development are (i) the short life of the reporter enabling to have a clear view of the onset as well as the termination of drug effects, (ii) the generalized, drug-dependent activation of the reporter, and (iii) imaging modality suitable for high-throughput analysis. Because of its relative cheapness and ease to perform, in addition to all the above considerations, bioluminescence-based imaging is now regarded as the best imaging technology to be applied to the field of drug research. We show here the application of reporter mouse systems for drug screening in living animals in order to compare drug potency on target and specificity of action. PMID- 20012394 TI - Human FcRn transgenic mice for pharmacokinetic evaluation of therapeutic antibodies. AB - Therapeutic monoclonal antibodies are widely recognized to be a most promising means to treat an increasing number of human diseases, including cancers and autoimmunity. To a large extent, the efficacy of monoclonal antibody treatment is because IgG antibodies have greatly extended persistence in vivo. However, conventional rodent models do not mirror human antibody pharmacokinetics. The key molecule responsible for the extended persistence antibodies is the major histocompatibility complex class I family Fc receptor, FcRn. We describe human FcRn transgenic mouse models and how they can be exploited productively for the preclinical pharmacokinetic evaluation of therapeutic antibodies. PMID- 20012395 TI - Development of novel major histocompatibility complex class I and class II deficient NOD-SCID IL2R gamma chain knockout mice for modeling human xenogeneic graft-versus-host disease. AB - Immunodeficient mice have been used as recipients of human peripheral blood mononuclear cells (PBMC) for in vivo analyses of human xeno-graft-versus-host disease (GVHD). This xeno-GVHD model system in many ways mimics the human disease. The model system is established by intravenous or intraperitoneal injection of human PBMC or spleen cells into unconditioned or irradiated immunodeficient recipient mice. Recently, the development of several stocks of immunodeficient Prkdc ( scid ) (scid) and recombination activating 1 or 2 gene (Rag1 or Rag2) knockout mice bearing a targeted mutation in the gene encoding the IL2 receptor gamma chain (IL2rgamma) have been reported. The addition of the mutated IL2rgamma gene onto an immunodeficient mouse stock facilitates heightened engraftment with human PBMC. Stocks of mice with mutations in the IL2rgamma gene have been studied in several laboratories on NOD-scid, NOD-Rag1 ( null ), BALB/c Rag1 ( null ), BALB/c-Rag2 ( null ), and Stock-H2(d)-Rag2 ( null ) strain backgrounds. Parameters to induce human xeno-GVHD in H2(d)-Rag2 ( null ) IL2rgamma ( null ) mice have been published, but variability in the frequency of disease and kinetics of GVHD were observed. The availability of the NOD-scid IL2rgamma ( null ) stock that engrafts more readily with human PBMC than does the Stock-H2(d)-Rag2 ( null ) IL2rgamma ( null ) stock should lead to a more reproducible humanized mouse model of GVHD and for the use in drug evaluation and validation. Furthermore, GVHD in human PBMC-engrafted scid mice has been postulated to result predominately from a human anti-mouse major histocompatibility complex (MHC) class II reactivity. Our recent development of NOD-scid IL2rgamma ( null ) beta2m ( null ) and NOD-scid IL2rgamma ( null ) Ab ( null ) stocks of mice now make it possible to investigate directly the role of host MHC class I and class II in the pathogenesis of GVHD in humanized mice using NOD-scid IL2rgamma ( null ) stocks that engraft at high levels with human PBMC and are deficient in murine MHC class I, class II, or both classes of MHC molecules. PMID- 20012396 TI - Bridging mice to men: using HLA transgenic mice to enhance the future prediction and prevention of autoimmune type 1 diabetes in humans. AB - Similar to the vast majority of cases in humans, the development of type 1 diabetes (T1D) in the NOD mouse model is due to T-cell mediated autoimmune destruction of insulin-producing pancreatic beta cells. Particular major histocompatibility complex (MHC) haplotypes (designated HLA in humans and H2 in mice) provide the primary genetic risk factor for T1D development. It has long been appreciated that within the MHC, particular unusual class II genes contribute to the development of T1D in both humans and NOD mice by allowing for the development and functional activation of beta-cell autoreactive CD4 T cells. However, studies in NOD mice have revealed that through interactions with other background susceptibility genes, the quite common class I variants (K(d), D(b)) characterizing this strain's H2 ( g7 ) MHC haplotype aberrantly acquire an ability to support the development of beta cell autoreactive CD8 T-cell responses also essential to T1D development. Similarly, recent studies indicate that in the proper genetic context some quite common HLA class I variants also aberrantly contribute to T1D development in humans. This chapter will focus on how "humanized" HLA transgenic NOD mice can be created and used to identify class I dependent beta cell autoreactive CD8 T-cell populations of clinical relevance to T1D development. There is also discussion on how HLA transgenic NOD mice can be used to develop protocols that may ultimately be useful for the prevention of T1D in humans by attenuating autoreactive CD8 T-cell responses against pancreatic beta cells. PMID- 20012397 TI - Mouse models of type II diabetes mellitus in drug discovery. AB - Type II diabetes is a fast-growing epidemic in industrialized countries. Many recent advances have led to the discovery and marketing of efficient novel therapeutic medications. Yet, because of side effects of these medications and the variability in individual patient responsiveness, unmet needs subsist for the discovery of new drugs. The mouse has proven to be a reliable model for discovering and validating new treatments for type II diabetes mellitus. We review here the most common mouse models used for drug discovery for the treatment of type II diabetes. The methods presented focus on measuring the equivalent end points in mice to the clinical values of glucose metabolism used for the diagnostic of type II diabetes in humans: i.e., baseline fasting glucose and insulin, glucose tolerance test, and insulin sensitivity index. Improvements on these clinical values are essential for the progression of a novel potential therapeutic molecule through a preclinical and clinical pipeline. PMID- 20012398 TI - Cholesterol absorption and metabolism. AB - Inhibitors of cholesterol absorption have been sought for decades as a means to treat and prevent cardiovascular diseases associated with hypercholesterolemia. Ezetimibe is the one clear success story in this regard, and other compounds with similar efficacy continue to be sought. In the last decade, the laboratory mouse, with all its genetic power, has become the premier experimental model for discovering the mechanisms underlying cholesterol absorption and has become a critical tool for preclinical testing of potential pharmaceutical entities. This chapter briefly reviews the history of cholesterol absorption research and the various gene candidates that have come under consideration as drug targets. The most common and versatile method of measuring cholesterol absorption is described in detail along with important considerations when interpreting results, and an alternative method is also presented. In recent years, reverse cholesterol transport has become an area of intense new interest for drug discovery since this process is now considered another key to reducing cardiovascular disease risk. The ultimate measure of reverse cholesterol transport is sterol excretion and a detailed description is given for measuring neutral and acidic fecal sterols and interpreting the results. PMID- 20012399 TI - Collagen-induced arthritis in mice. AB - Collagen-induced arthritis (CIA) in mice is an animal model for rheumatoid arthritis (RA) and can be induced in DBA/1 and C57BL/6 mice using different protocols. The CIA model can be used to unravel mechanisms involved in the development of arthritis and is frequently used to study the effect of new therapeutics. The development of a CIA model in C57BL/6 mice recently enabled researchers to use knockout mice on this background for arthritis research.In this chapter, the protocol for induction of arthritis in both mice strains is described, including the monitoring of clinical arthritis and paw swelling in the mice during the experiment. Furthermore, protocols for decalcification of paws and for the detection of collagen-specific antibodies in mice sera are described. PMID- 20012400 TI - Skin diseases in laboratory mice: approaches to drug target identification and efficacy screening. AB - A large variety of mouse models for human skin and adnexa diseases are readily available from investigators and vendors worldwide. While the skin is an obvious organ to observe lesions and their response to therapy, actually treating and monitoring progress in mice can be challenging. This chapter provides an overview on how to use the laboratory mouse as a preclinical tool to evaluate efficacy of a new compound or test potential new uses for a compound approved for use for treating an unrelated disease. Basic approaches to handling mice, applying compounds, and quantifying effects of the treatment are presented. PMID- 20012401 TI - Models of metastasis in drug discovery. AB - By definition, animal models provide only an approximation of clinical reality. One reason for this, for example, is that although metastases are the primary cause of mortality from neoplasia, by are rarely considered a target in drug discovery and development. Due to the impact of metastasis on clinical disease, we posit that metastasis should be considered in drug discovery, in addition, to more traditional biologic concepts, including drug pharmacology and toxicity. Drug discovery and developmental studies can incorporate orthotopic and spontaneous metastasis models (syngeneic and xenogeneic) with their inherent host tumor microenvironmental interactions, in addition to confirmatory autochthonous and/or genetically engineered models (GEMs). This requires a rational and hierarchical approach using models of metastatic disease optimally using resected, orthotopic primary tumors and clinically relevant outcome parameters. In this chapter, we provide protocols for models of metastasis that can be used in translational and drug discovery studies. PMID- 20012402 TI - Methods for evaluating effects of an irinotecan + 5-fluorouracil/leucovorin (IFL) regimen in an orthotopic metastatic colorectal cancer model utilizing in vivo bioluminescence imaging. AB - In testing novel anticancer therapies, researchers strive to utilize models that reflect the human disease as much as feasible. In this regard, orthotopic models are frequently developed because cancer cells in these models form tumors in, and metastasize from, a tissue environment similar to the tissue of origin of the cancer cells. Here we adapted an orthotopic colorectal cancer model, in which HT 29 colorectal cancer cells form tumors in the rectal lining and metastasize to the para-aortic lymph nodes with high frequency. Firefly luciferase-expressing HT 29 cells were used in this model to realize the benefits of bioluminescence imaging (BLI). A combination of irinotecan, 5-fluorouracil (5-FU), and leucovorin (LV) (IFL) was used as a standard chemotherapeutic regimen positive control. BLI allowed for the demonstration of the effects of IFL on tumor growth in the rectal lining, with tumor weight measurements at the end of the study reflecting total tumor burden. BLI also allowed relatively easy demonstration of reduced tissue metastasis with IFL treatment, compared to more time-consuming histological techniques. It is concluded that the orthotopic colorectal cancer model approach described represents a valuable tool for validating treatment strategies in this indication. PMID- 20012403 TI - CML mouse model in translational research. AB - Chronic myeloid leukemia (CML) is a myeloproliferative disorder characterized by increased proliferation of granulocytic cells without the loss of their capability to differentiate. CML is derived from the hematopoietic stem cells (1) with the Philadelphia chromosome resulting from of a reciprocal translocation between the chromosomes 9 and 22 t(9;22)-(q34;q11). This translocation produces a fusion gene known as BCR-ABL which acquires uncontrolled tyrosine kinase activity, constantly turning on its downstream signaling molecules/pathways, and promoting proliferation of leukemia cell through anti-apoptosis and acquisition of additional mutations. To evaluate the role of each critical downstream signaling molecule of BCR-ABL and test therapeutic drugs in vivo, it is important to use physiological mouse disease models. In this chapter, we describe a mouse model of CML induced by BCR-ABL retrovirus (MSCV-BCR-ABL-GFP; MIG-BCR-ABL) and how to use this model in translational research. PMID- 20012404 TI - Mouse models for studying depression-like states and antidepressant drugs. AB - Depression is a common psychiatric disorder, with diverse symptoms and high comorbidity with other brain dysfunctions. Due to this complexity, little is known about the neural and genetic mechanisms involved in depression pathogenesis. In a large proportion of patients, current antidepressant treatments are often ineffective and/or have undesirable side effects, fueling the search for more effective drugs. Animal models mimicking various symptoms of depression are indispensable in studying the biological mechanisms of this disease. Here, we summarize several popular methods for assessing depression-like symptoms in mice and their utility in screening antidepressant drugs. PMID- 20012405 TI - Virus-delivered RNA interference in mouse brain to study addiction-related behaviors. AB - The use of viral vectors for gene transfer to specific brain regions is a powerful tool for determining gene function in mouse behavioral models. We have employed a lentiviral vector to deliver small-hairpin RNAs to areas of mouse brain implicated in behaviors relevant to drug addiction, such as the nucleus accumbens and ventral tegmental area. Delivery of virus expressing small-hairpin RNAs results in sustained target gene knockdown by RNA interference. Mice can subsequently be tested for behavioral responses to various drugs of abuse over the course of several weeks or months. Here we describe a method for stereotaxic delivery of lentivirus to mouse brain. This method is widely applicable to any behavioral experiment in which the role of a specific gene in a particular brain region is to be elucidated. PMID- 20012406 TI - Experimental models of anxiety for drug discovery and brain research. AB - Animal models have been vital to recent advances in experimental neuroscience, including the modeling of common human brain disorders such as anxiety, depression, and schizophrenia. As mice express robust anxiety-like behaviors when exposed to stressors (e.g., novelty, bright light, or social confrontation), these phenotypes have clear utility in testing the effects of psychotropic drugs. Of specific interest is the extent to which mouse models can be used for the screening of new anxiolytic drugs and verification of their possible applications in humans. To address this problem, the present chapter will review different experimental models of mouse anxiety and discuss their utility for testing anxiolytic and anxiogenic drugs. Detailed protocols will be provided for these paradigms, and possible confounds will be addressed accordingly. PMID- 20012407 TI - Mouse models of neurodegenerative diseases: criteria and general methodology. AB - The major symptom of Alzheimer's disease is rapidly progressing dementia, coinciding with the formation of amyloid and tau deposits in the central nervous system, and neuronal death. At present familial cases of dementias provide the most promising foundation for modelling neurodegeneration. We describe the mnemonic and other major behavioral symptoms of tauopathies, briefly outline the genetics underlying familiar cases and discuss the arising implications for modelling the disease in mostly transgenic mouse lines. We then depict to what degree the most recent mouse models replicate pathological and cognitive characteristics observed in patients.There is no universally valid behavioral test battery to evaluate mouse models. The selection of individual tests depends on the behavioral and/or memory system in focus, the type of a model and how well it replicates the pathology of a disease and the amount of control over the genetic background of the mouse model. However it is possible to provide guidelines and criteria for modelling the neurodegeneration, setting up the experiments and choosing relevant tests. One should not adopt a "one (trans)gene, one disease" interpretation, but should try to understand how the mouse genome copes with the protein expression of the transgene in question. Further, it is not possible to recommend some mouse models over others since each model is valuable within its own constraints, and the way experiments are performed often reflects the idiosyncratic reality of specific laboratories. Our purpose is to improve bridging molecular and behavioural approaches in translational research. PMID- 20012408 TI - Neuromuscular disease models and analysis. AB - Neuromuscular diseases can affect the survival of peripheral neurons, their axons extending to peripheral targets, their synaptic connections onto those targets, or the targets themselves. Examples include motor neuron diseases such as amyotrophic lateral sclerosis, peripheral neuropathies, such as Charcot-Marie Tooth diseases, myasthenias, and muscular dystrophies. Characterizing these phenotypes in mouse models requires an integrated approach, examining both the nerve and the muscle histologically, anatomically, and functionally by electrophysiology. Defects observed at these levels can be related back to onset, severity, and progression, as assessed by "quality-of-life measures" including tests of gross motor performance such as gait or grip strength. This chapter describes methods for assessing neuromuscular disease models in mice, and how interpretation of these tests can be complicated by the inter-relatedness of the phenotypes. PMID- 20012409 TI - Murine model of cutaneous infection with Streptococcus pyogenes. AB - Despite the medical advances achieved during the last century to fight against bacteria, viruses, fungi and parasites, infectious diseases are still a major cause of death, disability, and social and economic upheaval for millions around the world. Challenges remain in countering microorganisms even where antibiotics and vaccines are available. Much remains to be learned about basic aspects of the host-pathogen relationship and the complexity of the immune response to infection. Animal models represent a powerful tool to dissect the host response to infection, as well as the pathogenesis of the microbe. One of the advantages of using animal models is that both genetic and environmental factors that may influence the course of an infection can be controlled, allowing a precise cause effect analysis of the host-pathogen interactions. In addition, there are no real alternatives to whole animal models in the study of integrative physiology and dynamic pathophysiologic alterations. The use of animal models has also proven invaluable for testing the efficacy of experimental antimicrobial agents and their therapeutic regimes. The mouse model is the most widely used for many reasons, including its cost effectiveness, the high number of immunological reagents available for this species, and the relative ease of biocontainment. Mouse strains with specific properties such as transgenic mouse strains with gene insertion or targeted mutation (knock-out) are very effective tools for studying the role of specific genes controlling the immune response to infectious pathogens. Murine models will remain the most appropriate tool for evaluating new therapeutic strategies for the treatment of various diseases. The closer the model is adapted to the human disease, the more reliable will be the results.In this chapter, the experimental procedures required to establish a mouse model of cutaneous and soft tissue infection are detailed. This model has provided invaluable insights into the pathogenicity of the agent for the human host. PMID- 20012410 TI - Murine model of pneumococcal pneumonia. AB - Respiratory tract infections remain among the most common clinical problems worldwide. Pneumonia or inflammation of the lungs can be caused by infection with bacteria, viruses, and other organisms. Pneumonia management has been challenged by the widespread distribution of antibiotic-resistant strains of Streptococcus pneumoniae, the commonest cause of community acquired pneumonia. Experimental models of pneumonia have played a crucial role for testing the efficacy of antimicrobial agents as well as for gaining a better understanding of the disease pathogenesis. These models have also received increased attention as tools for deriving pharmacodynamic data and for determining the clinical significance of drug resistance. PMID- 20012411 TI - Murine model of polymicrobial septic peritonitis using cecal ligation and puncture (CLP). AB - Although a number of animal models such as endotoxic shock and bacteremia have been used to study the pathogenesis of sepsis, cecal ligation and puncture (CLP) represents a peritonitis model with clinical features of polymicrobial infection comparable with those of peritonitis in humans. The CLP consists in the surgical perforation of the legated cecum of mice that results in immediate and constant drainage of cecal bacteria into the peritoneal cavity. The severity of the diseases depends on the diameter of the needle used for the perforation as well as on the number of cecal punctures. The CLP model of sepsis in mice is the most commonly used for studying the process of septic peritonitis and can be used as a preclinical model to test the efficacy of pharmacological agents for the treatment of sepsis. PMID- 20012412 TI - G-quadruplexes: from guanine gels to chemotherapeutics. AB - G-quartets are square planar arrangements of four guanine bases, which can form extraordinarily stable stacks when present in nucleic acid sequences. Such G quadruplex structures were long regarded as an in vitro phenomenon, but the widespread presence of suitable sequences in genomes and the identification of proteins that stabilize, modify, or resolve these nucleic acid structures have provided circumstantial evidence for their physiological relevance. The therapeutic potential of small molecules that can stabilize or disrupt G quadruplex structures has invigorated the field in recent years. Here we review some of the key observations that support biological functions for G-quadruplex DNA as well as the techniques and tools that have enabled researchers to probe these structures and their interactions with proteins and small molecules. PMID- 20012413 TI - Molecular modeling and simulation of G-quadruplexes and quadruplex-ligand complexes. AB - Methods for the molecular modeling and simulation of G-quadruplex structures and their drug/ligand complexes are discussed, and a range of protocols is presented for undertaking a variety of tasks including model-building, ligand docking, dynamics simulation, continuum solvent modeling, energetic calculations, principal component analysis, and quantum chemical computations. The scope and limitations of these approaches are discussed. PMID- 20012414 TI - Computational approaches to the detection and analysis of sequences with intramolecular G-quadruplex forming potential. AB - Sequences with the potential to form intramolecular G-quadruplexes (G4 structures) are found in highly nonrandom distributions in the genomes of diverse organisms. These sequences are associated with nucleic acid metabolic processes ranging from transcription and translation to recombination and telomere function. Here we review different computational methods for identifying potential G4-forming sequences and provide protocols for their implementation. We also discuss methods for assessing the significance and specificity of associations between the sequences and different biological functions. PMID- 20012415 TI - Preparation of G-quartet structures and detection by native gel electrophoresis. AB - Mounting evidence supporting the existence of DNA structures containing G quartets in vivo makes these unique and diverse nucleic acid structures an important research subject, and future investigations aimed at elucidating their biological significance are expected. The purification and characterization of G quartet structures can be challenging because their inherent structural diversity, complexity, and stability are sensitive to an array of variables. The stability of G-quartet structures depends on many factors including number of DNA strands involved in G-quartet formation, the identity of the stabilizing cation(s), the number and sequence context of the guanosines involved in stacking, the presence of single-stranded overhangs, the intervening loop size, and the identity of nucleosides in the loop. Here we detail current methods used in G-quartet preparation and their purification and characterization by native gel electrophoresis. PMID- 20012417 TI - Real-time observation of G-quadruplex dynamics using single-molecule FRET microscopy. AB - The potential importance of G-quadruplex structures was implied by the recent findings that the human POT1 disrupts G-quadruplex and stimulates the telomerase activity. A solid understanding of the range of conformations that can be adopted by guanine-rich sequences can potentially shed much light on the molecular mechanisms underlying certain human diseases related to telomeres. Furthermore, structure-based design of chemotherapeutic drugs for cancer might be realized by addressing different types of G-quadruplex structures. Using the unique capabilities of single-molecule spectroscopy, we have recently reported on the intricate dynamic structural properties of a minimal form of human telomeric DNA. Here, we present the detailed step-by-step methods for the real-time observation of G-rich DNA sequences by means of single-molecule FRET microscopy and provide the protocols for vesicle encapsulation and surface immobilization assays. Such assays provide a firm basis for future studies aimed at elucidating the interaction between telomeric DNA and telomere-associated proteins as well as the synthetic therapeutic agents that specifically stabilize certain G-quadruplex topologies. PMID- 20012416 TI - Biochemical techniques for the characterization of G-quadruplex structures: EMSA, DMS footprinting, and DNA polymerase stop assay. AB - The proximal promoter region of many human growth-related genes contains a polypurine/polypyrimidine tract that serves as multiple binding sites for Sp1 or other transcription factors. These tracts often contain a guanine-rich sequence consisting of four runs of three or more contiguous guanines separated by one or more bases, corresponding to a general motif known for the formation of an intramolecular G-quadruplex. Recent results provide strong evidence that specific G-quadruplex structures form naturally within these polypurine/polypyrimidine tracts in many human promoter regions, raising the possibility that the transcriptional control of these genes can be modulated by G-quadruplex interactive agents. In this chapter, we describe three general biochemical methodologies, electrophoretic mobility shift assay (EMSA), dimethylsulfate (DMS) footprinting, and the DNA polymerase stop assay, which can be useful for initial characterization of G-quadruplex structures formed by G-rich sequences. PMID- 20012418 TI - Sedimentation velocity ultracentrifugation analysis for hydrodynamic characterization of G-quadruplex structures. AB - Analytical ultracentrifugation (AUC) is a powerful technique for the characterization of hydrodynamic and thermodynamic properties. The intent of this article is to demonstrate the utility of sedimentation velocity (SV) studies to obtain hydrodynamic information for G-quadruplex (GQ) systems and to provide insights into one part of this process, namely, data analysis of existing SV data. An array of data analysis software is available, mostly written and continually developed by established researchers in the AUC field, with particularly rapid advances in the analysis of SV data. Each program has its own learning curve, and this article is intended as a resource in the data analysis process for beginning researchers in the field. We discuss the application of three of the most commonly used data analysis programs, DCDT+, Sedfit, and SedAnal, to the interpretation of SV data obtained in our laboratory on two GQ systems. PMID- 20012419 TI - 2-aminopurine as a probe for quadruplex loop structures. AB - Fluorescent reporter groups have served for many years as sensitive probes of macromolecular structure. Such probes can be especially useful in comparative studies such as detection of conformational changes and discrimination among structural models. Spectroscopic methods such as fluorescence are attractive because they are rapid, require small amounts of material, are nondestructive, can be carried out with commonly available equipment, and are relatively inexpensive. In addition, there is a rich library of theoretical and practical materials available to aid in data interpretation.The intrinsic fluorescence of most nucleic acids is too low to be useful in structural studies. Thus, it is necessary to incorporate a suitable reporter group to utilize fluorescence methods involving polynucleotide structure. A highly fluorescent adenine analog, 2-aminopurine, has long served in this capacity. The present article describes our use of 2-aminopurine as a probe of loop structures in quadruplex DNA. In particular, we show how knowledge of the relative intensity of 2-aminopurine emission as well as its sensitivity to exogenous quenching molecules such as acrylamide can aid in comparing crystal and solution structures of an oligonucleotide model of the human telomere and in discrimination among models containing tandem repeats of the telomeric quadruplex. PMID- 20012420 TI - Assessing DNA structures with 125I radioprobing. AB - Iodine-125 radioprobing is based on incorporation of radioiodine into a defined position in a nucleic acid molecule. Decay of (125)I results in the emission of multiple, low-energy Auger electrons that, along with positively charged residual daughter nuclide, produce DNA strand breaks. The probability of such strand breaks at a given nucleotide is in inverse proportion to the distance from the (125)I atom to the sugar of that nucleotide. Therefore, conclusions can be drawn about the conformation or folding of a DNA or RNA molecule based on the distribution of (125)I decay-induced strand breaks. Here we describe in detail the application (125)I radioprobing for studying the conformation of quadruplex structures, and discuss the advantages and limitations of the method. PMID- 20012421 TI - Monitoring the temperature unfolding of G-quadruplexes by UV and circular dichroism spectroscopies and calorimetry techniques. AB - DNA oligonucleotides containing guanine repeat sequences can adopt G-quadruplex (GQ) structures in the presence of specific metal ions. We report on how to use a combination of spectroscopic and calorimetric techniques to determine the spectral characteristics and thermodynamic parameters for the temperature unfolding of GQs. Specifically, we investigated the unfolding of d(G(2)T(2)G(2)TGTG(2)T(2)G(2)), G2, and d(G(3)T(2)G(3)TGTG(3)T(2)G(3)), G3 by a combination of UV and circular dichroism (CD) spectroscopies, and differential scanning calorimetry (DSC).Analysis of the UV and CD spectra of these GQs at low (100% helix) and high (100% random coil) temperatures yielded the optimal wavelengths to determine the melting curves. In addition, the CD spectra yielded the particular conformation(s) that each GQ adopted at low temperature. DSC curves yielded complete thermodynamic profiles for the unfolding of each GQ. We use these profiles to determine the thermodynamic contributions for the formation of a G-quartet stack. PMID- 20012422 TI - Probing telomeric G-quadruplex DNA structures in cells with in vitro generated single-chain antibody fragments. AB - Guanine-rich sequences have been shown to readily form parallel or antiparallel G quadruplex DNA structures in vitro. All telomeric repeat sequences contain stretches of guanine residues that can form quadruplex structures. In order to demonstrate the occurrence of the quadruplex structure in vivo, we generated by ribosome display, scFv antibodies specific for quadruplex DNA structures formed by the telomeric sequence of the ciliate Stylonychia. The macronucleus of this hypotrichous ciliate contains 10(8) telomere-capped nanochromosomes and was stained with the antibody recognizing the antiparallel G-quadruplex DNA in indirect immuno-fluorescence assays. This antibody was also used as a specific probe to study the interaction of the telomere end-binding proteins with the G quadruplex during different stages of the cell cycle. PMID- 20012423 TI - Detection of G-quadruplexes in cells and investigation of G-quadruplex structure of d(T2AG3)4 in K+ solution by a carbazole derivative: BMVC. AB - Verification of the existence of quadruplex structure in native human telomeres and determination of the major structure of d(T(2)AG(3))(4) (H24) in K(+) solution are the major questions regarding the structure of human telomeres. We have synthesized a fluorescent probe of 3,6-bis(1-methyl-4 vinylpyridinium)carbazole diiodide (BMVC) that has a very high binding affinity for G-quadruplex H24. BMVC stabilizes quadruplex structures and acts as a sensitive probe to the local environment. Although the circular dichroism patterns of H24 are different in Na(+) and K(+) solutions, similar binding behaviors of BMVC to H24 in these solutions led us to suggest that the major G quadruplex structure of H24 in K(+) solution is very likely similar to that in Na(+) solution. Of particular interest is the fluorescent band detected at -575 nm in quadruplex H24 and at -545 nm in duplex DNA. In addition, the intensity of BMVC fluorescence increases by two orders of magnitudes upon interaction with either duplex or G-quadruplex DNA. BMVC has a greater binding preference for G quadruplex H24 than for duplex DNA. Analyzing the BMVC fluorescence at the ends of metaphase chromosomes and other regions of chromosomes allowed us to verify the presence of G-quadruplex structure in human telomeres for the first time. Using fluorescence lifetime imaging microscopy, the longer decay time of BMVC in G-quadruplex H24 than in duplex DNA allowed us to map the G-quadruplex structure in human metaphase chromosomes. PMID- 20012424 TI - Isolation of G-quadruplex DNA using NMM-sepharose affinity chromatography. AB - DNA can adopt a variety of non-standard conformations, including structures known as G-quadruplexes (G4-DNA), which consist of stacked tetrads of guanines. There are growing indications that G4-DNA is of biological importance, including evidence that it plays roles in telomere function, DNA recombination and the regulation of transcription and translation. However, it has been difficult to obtain direct, physical evidence for the presence of G-quadruplex DNA in vivo due, in part, to a lack of tools for G4-DNA identification. Here, we describe a method for coupling the G4-DNA binding ligand N-methyl mesoporphyrin IX (NMM) to a Sepharose resin, and demonstrate the ability of the resin to bind tightly and selectively to DNA oligonucleotides with the capacity to form G4-DNA. This technique might also be extended to examine genomic distributions of G4-DNA isolated from in vivo sources. PMID- 20012425 TI - Quantifying interactions between G-quadruplex DNA and transition-metal complexes. AB - Telomerase inhibition through guanine quadruplex sequestration by small-molecule drugs is of great current interest as an anticancer strategy. G-quadruplexes (GQs) can be formed at the guanine-rich sequences found at the end of the telomere. They possess a large electron-rich pi-surface which is favorable for the binding of electron-poor small molecules. Small molecules binding to GQs can sequester the telomere ends and inhibit the enzyme telomerase, which is expressed in cancer cells and absent in normal somatic cells. Transition-metal complexes present a myriad of geometries and numerous ligand coordination environments and allow for modular syntheses for development of compound libraries to target GQs. We have demonstrated the size of the pi-surface, binding selectivity and affinity of phenanthroimidazole platinum (II) complexes [PtPIX(en)](2+)2PF (6) (-) (X = naphthyl, phenyl and en = ethylenediamine) and metallosupramolecular complexes [Pt(4,4'-bpy)(en)] (4) (8+) 8PF (6) (-) (where bpy = bipyridine) to GQs can be readily tuned and assayed through a number of biophysical techniques. PMID- 20012426 TI - G4-FID: a fluorescent DNA probe displacement assay for rapid evaluation of quadruplex ligands. AB - Currently, small molecules able to interact specifically with G-quadruplex DNA (G quadruplex ligands) are intensively studied since they appear to inhibit the growth of cancer cells via an unprecedented mode of action based on structural perturbation of telomeres. It is thus of crucial interest to develop methods that enable easy, rapid and reliable detection of valuable candidates. Herein, we would like to report on the G4-FID assay, a simple fluorescence screening method based on the displacement of a DNA light-up probe (thiazole orange) from both quadruplex and duplex DNA, therefore allowing a concomitant evaluation of the quadruplex-affinity and quadruplex- over duplex-selectivity of the tested candidates. PMID- 20012427 TI - [Ultrasound-guided central venous access in adults and children: Procedure and pathological findings]. AB - Central venous line placement is a standard procedure in critical care and peri operative medicine. This procedure can be associated with severe complications. In contrast to the landmark technique, ultrasound-guided punctures can significantly reduce the rate of complications. Patients with a high risk for difficult vascular access include critical care and emergency patients as well as patients on anticoagulation medication and dialysis. Placement of central venous catheters can be difficult in ventilated patients and if there has been prior surgery in the puncture area. In children and small infants central venous access can also be challenging due to the anatomical relationship in the head and neck region. Puncture techniques are explained briefly by means of ultrasound anatomy. Typical ultrasonographic images visualize pathological findings in order to identify dangers and complications in central venous catheterization. PMID- 20012428 TI - [Partial weight-bearing in rehabilitation. Strategies for instruction and limitations]. AB - Following trauma or surgery on the musculoskeletal system the primary aim is always as complete a restitution of mobility as possible. By mobilization with partial weight-bearing this is possible. The preferred way of teaching partial weight-bearing is the use of conventional bathroom scales. This method proves to be simple as well as time and cost-saving, but the transferability to the patient's daily life is questionable. Training and control of partial weight bearing under dynamic conditions, such as normal walking, and walking up and down stairs seem to be very important. Different investigations have shown that the minority of subjects recruited could manage to maintain the given load of partial weight-bearing. Furthermore, the actual resulting moments within the joints, caused by muscles, fascia and tendons, are not considered in presets of partial weight-bearing, as only external forces (ground reaction forces) are measured. However, the problems in teaching partial weight-bearing have to be contrasted with the as yet unexplained issue of postoperative partial versus full weight bearing. PMID- 20012429 TI - Primary versus recurrent prolapse surgery: differences in outcomes. AB - INTRODUCTION AND HYPOTHESIS: We set out to evaluate anatomical outcomes of recurrent vs. primary prolapse surgery, focusing on anterior colporrhaphy (AC). METHODS: A retrospective study was performed comparing patients who underwent AC for recurrent cystocele (group I) and a matched control group who underwent primary AC (group II). RESULTS: Thirty-one patients were included in each group. Median follow-up was 22 (5-55) months. Successful anterior vaginal support was obtained in 18/23 (78.2%) patients in group I and 17/21 (81%) patients in group II at 1 year (p = 1.000) and in 9/21 (42.8%) patients in group I and in 15/21 (71.4%) patients in group II at 2-year follow-up (p = 0.031). CONCLUSIONS: Recurrent cystocele repair has a higher anatomic failure rate than primary repair at 2-year follow-up. Alternative surgical techniques that provide better long term durability may be beneficial in repair of recurrent anterior wall prolapse. PMID- 20012431 TI - In vivo hemodynamic analysis of intracranial aneurysms obtained by magnetic resonance fluid dynamics (MRFD) based on time-resolved three-dimensional phase contrast MRI. AB - INTRODUCTION: Hemodynamics is thought to play a very important role in the initiation, growth, and rupture of intracranial aneurysms. The purpose of our study was to perform in vivo hemodynamic analysis of unruptured intracranial aneurysms of magnetic resonance fluid dynamics using time-resolved three dimensional phase-contrast MRI (4D-Flow) at 1.5 T and to analyze relationships between hemodynamics and wall shear stress (WSS) and oscillatory shear index (OSI). METHODS: This study included nine subjects with 14 unruptured aneurysms. 4D-Flow was performed by a 1.5-T magnetic resonance scanner with a head coil. We calculated in vivo streamlines, WSS, and OSI of intracranial aneurysms based on 4D-Flow with our software. We evaluated the number of spiral flows in the aneurysms and compared the differences in WSS or OSI between the vessel and aneurysm and between whole aneurysm and the apex of the spiral flow. RESULTS: 3D streamlines, WSS, and OSI distribution maps in arbitrary direction during the cardiac phase were obtained for all intracranial aneurysms. Twelve aneurysms had one spiral flow each, and two aneurysms had two spiral flows each. The WSS was lower and the OSI was higher in the aneurysm compared to the vessel. The apex of the spiral flow had a lower WSS and higher OSI relative to the whole aneurysm. CONCLUSION: Each intracranial aneurysm in this study had at least one spiral flow. The WSS was lower and OSI was higher at the apex of the spiral flow than the whole aneurysmal wall. PMID- 20012430 TI - In vitro-in vivo extrapolation of zolpidem as a perpetrator of metabolic interactions involving CYP3A. AB - OBJECTIVES: To evaluate zolpidem as a mechanism-based inactivator of human CYP3A in vitro, and to assess its metabolic interaction potential with CYP3A drugs (in vitro-in vivo extrapolation; IV-IVE). METHODS: A co- vs. pre-incubation strategy was used to quantify time-dependent inhibition of human liver microsomal (HLM) and recombinant CYP3A4 (rCYP3A4) by zolpidem. Experiments involving a 10-fold dilution step were employed to determine the kinetic constants of inactivation (K (I) and k (inact)) and to assess the in vitro mechanism-based inactivation (MBI) criteria. Inactivation data were entered into the Simcyp population-based ADME simulator to predict the increase in the area under the plasma concentration-time curve (AUC) for orally administered midazolam. RESULTS: Consistent with MBI, the inhibitory potency of zolpidem toward CYP3A was increased following pre incubation. In HLMs, the concentration required for half maximal inactivation (K (I)) was 122 microM and the maximal rate of inactivation (k (inact)) was 0.094 min(-1). In comparison, K (I) and k (inact) values with rCYP3A4 were 50 microM and 0.229 min(-1), respectively. Zolpidem fulfilled all other in vitro MBI criteria, including irreversible inhibition. The mean oral AUC for midazolam in healthy volunteers was predicted to increase 1.1- to 1.7-fold due to the inhibition of metabolic clearance by zolpidem. Elderly subjects were more sensitive to the interaction, with mean increases in midazolam AUC of 1.2- and 2.2-fold for HLM IV-IVE and rCYP3A4 IV-IVE, respectively. CONCLUSIONS: Zolpidem is a relatively weak mechanism-based inactivator of human CYP3A in vitro. Zolpidem is unlikely to act as a significant perpetrator of metabolic interactions involving CYP3A. PMID- 20012432 TI - Effect of high aluminum concentration in water resources on human health, case study: Biga Peninsula, northwest part of Turkey. AB - Widespread and intense zones of silicified, propylitic, and argillic alteration exist as outcrop around the Biga Peninsula, NW Turkey. Most of the springs in the study area surface out from these altered volcanic rocks. The concentrations of aluminum (Al) in these springs ranged from 13.17 to 15.70 ppm in this region. These high levels of Al were found to exceed the maximum allowable limits (0.2 ppm) depicted in national and international standards of drinking water quality. Therefore, the effect of high Al in water resources on human health was evaluated in this research. A total of 273 people aged above 18 years and living in the Kirazli region (whose water supply is from springs emerging from these alteration zones) and in the Ciplak-Halileli region (whose water supply is provided from an alluvium aquifer) were selected as the research group. For this group, a questionnaire was completed that contained questions on descriptive characteristics of humans and a Mini-Mental State Examination (MMSE) was administered by the authors using the face-to-face interview technique. A neurological examination was then performed by the neurology specialist as a second-stage investigation. Finally, 10 ml of venous blood samples were obtained from these people as a third-stage analysis to determine the serum Al levels together with vitamin B(12), folic acid, and thyroid-stimulating hormone parameters. The result typically revealed that the MMSE score was less in 31.9% and there was no statistically significant difference between the two regions. However, the result also showed that neuropathy in the history (including a careful past medical history) was significantly higher in the Kirazli region. PMID- 20012433 TI - Application of a shape-memory alloy internal fixator for treatment of acetabular fractures with a follow-up of two to nine years in China. AB - Displaced acetabular fractures should be treated surgically. Over the past decade, surgical approaches to the acetabulum and the surgical technique for repair of common fracture patterns have advanced. Excellent outcomes after repair of these injuries can be achieved. The aim of this study was to assess the medium term results of reconstruction of acetabular fractures by using shape-memory alloy designed by the authors. This is a retrospective review conducted at a level 1 trauma centre. From October 1999 to July 2009, 19 patients with acetabular fractures were treated with our patented Ni-Ti shape-memory alloy acetabular tridimensional memory alloy-fixation system (ATMFS). The ATMFS device was cooled with ice before implantation and then warmed to 40-50 degrees C after implantation to produce balanced axial and compression forces that would stabilise the fracture three dimensionally. Our results are as follows; according to the D'Aubigne-Postel scoring system: Fifteen cases out of 19 (79%) achieved excellent or good clinical results. In two patients, late complications included avascular necrosis of the femoral head (ANFH) associated with posterior dislocation of the hip joint two years after the operation. We also observed two cases of grade II or III ectopic ossification, with good hip function, and one case of traumatic arthritis. In conclusion, these results demonstrate the effectiveness of the ATMFS device for the management of acetabular fracture. The device provides continuous compression of the fracture with minimal disruption to the local blood supply. PMID- 20012434 TI - The clinicopathological analysis of 303 cases with malignant lymphoma classified according to the World Health Organization classification system in a single institute of Taiwan. AB - Several reports have shown a different distribution of malignant lymphoma (ML) in Asian and Western populations. The purpose of our survey was to elucidate whether there are substantial differences in the frequencies of subtypes of ML between different geographical areas. All entities diagnosed as ML between June 1995 and December 2007 were selected according to the 2008 World Health Organization (WHO) classification and searched for clinical outcomes. The cases were retrieved and reviewed by a panel of clinical haematologists and haematopathologists. A total of 303 patients with ML were identified for retrospective analysis. Of the 303 patients with ML, 278 patients (91.7%) had non-Hodgkin's lymphoma (NHL), and 25 (9.2%) had Hodgkin's lymphoma. Of the 278 patients with NHL, 223 (73.6%) had lymphoma of B-cell lineage, and 55 (18.1%) had lymphoma of T-cell lineage. One hundred and thirty-seven patients were diagnosed with diffuse large B-cell lymphoma, which was the most common B-cell lineage subtype and accounted for 45.2% of patients with NHL. Peripheral T-cell lymphomas were the most frequent subset of the T-cell neoplasms, comprising 10.6% of ML. Extranodal involvement was found in 125 (44.9%) of the 278 patients with NHL, and the lymph node was the site of primary involvement in 153 patients (55.1%). Fifty-nine (47.2%) of the 125 patients with extranodal presentation had gastrointestinal tract involvement. Outcome was worse in patients with extranodal NHL than in those with nodal NHL through the entire follow-up period; the difference in survival rates was significant. Our findings clarify the applicability and prognostic relevance of the WHO classification system and provide further information about the incidence of various lymphoma subtypes in Taiwan. Primary extranodal NHL was associated with a worse prognosis and distinct characteristics compared with nodal NHL. The outcome of different types of extranodal NHL should be investigated further. PMID- 20012435 TI - Effects of hypothyroidism on myosin heavy chain composition and fibre types of fast skeletal muscles in a small marsupial, Antechinus flavipes. AB - Effects of drug-induced hypothyroidism on myosin heavy chain (MyHC) content and fibre types of fast skeletal muscles were studied in a small marsupial, Antechinus flavipes. SDS-PAGE of MyHCs from the tibialis anterior and gastrocnemius revealed four isoforms, 2B, 2X, 2A and slow, in that order of decreasing abundance. After 5 weeks treatment with methimazole, the functionally fastest 2B MyHC significantly decreased, while 2X, 2A and slow MyHCs increased. Immunohistochemistry using monospecific antibodies to each of the four MyHCs revealed decreased 2b and 2x fibres, and increased 2a and hybrid fibres co expressing two or three MyHCs. In the normally homogeneously fast superficial regions of these muscles, evenly distributed slow-staining fibres appeared, resembling the distribution of slow primary myotubes in fast muscles during development. Hybrid fibres containing 2A and slow MyHCs were virtually absent. These results are more detailed but broadly similar to the earlier studies on eutherians. We hypothesize that hypothyroidism essentially reverses the effects of thyroid hormone on MyHC gene expression of muscle fibres during myogenesis, which differ according to the developmental origin of the fibre: it induces slow MyHC expression in 2b fibres derived from fast primary myotubes, and shifts fast MyHC expression in fibres of secondary origin towards 2A, but not slow, MyHC. PMID- 20012437 TI - New experimental corrosive esophagitis model in rats. AB - PURPOSE: Caustic esophagitis is a serious clinical problem and many agents are currently tried out in many experimental models. The model of Gehanno is the most commonly used invasive model, which is required general anesthesia and laparotomy. We aimed to form a new pratic and non-invasive model. METHODS: Twenty rats were studied. The stomachs of the rats were reached through guidance catheter with ether anesthesia, Fogarty catheter was send in through, it was filled with pressure in the stomach. Then, Fogarty was pulled back and stomach entrance was closed. Control group was given; n = 10; 0.25 cc isotonic, injury group was given; n = 10; 0.25 cc, %40 NaOH and it was waited for 60 s. Their esophagi were examined after 28 days. RESULTS: In the histopathologic evaluation of the control group, no pathology was discovered. Sub-mucosal collagen increase, muscularis mucosa and tunica muscularis damage have all been detected in the injury group p < 0.005; p < 0.003; p < 0.005). CONCLUSIONS: Corrosive esophagitis was formed without general anesthesia and laparotomy. Burn was formed in the total esophagus, unlike other models in which the burn is just formed at the below end. With our less invasive, more easily applied model; treatment agents can be given just as the corrosive esophagitis can be formed. PMID- 20012436 TI - Calcium transport and homeostasis in gill cells of a freshwater crab Dilocarcinus pagei. AB - Crustaceans present a very interesting model system to study the process of calcification and calcium (Ca(2+)) transport because of molting-related events and the deposition of CaCO(3) in the new exoskeleton. Dilocarcinus pagei, a freshwater crab endemic to Brazil, was studied to understand Ca(2+) transport in whole gill cells using a fluorescent probe. Cells were dissociated, all of the gill cell types were loaded with fluo-3 and intracellular Ca(2+) change was monitored by adding Ca as CaCl(2) (0, 0.1, 0.25, 0.50, 1.0 and 5 mM), with a series of different inhibitors. For control gill cells, Ca(2+) transport followed Michaelis-Menten kinetics with K(m) = 0.42 +/- 0.04 mM and V(max) = 0.50 +/- 0.02 microM (Ca(2+) change x initial intracellular Ca(-1) x 180 s(-1); N = 14, r (2) = 0.99). Verapamil (a Ca(2+) channel inhibitor) and amiloride (a Na(+)/Ca(2+) exchanger [NCX] inhibitor) completely reduced intracellular Ca(2+) transport, while nifedipine, another Ca(2+) channel inhibitor, did not. Vanadate, a plasma membrane Ca(2+)-ATPase inhibitor (PMCA), increased intracellular Ca(2+) in gill cells through a decrease in the efflux of Ca(2+). Ouabain increased intracellular Ca(2+), similar to the effect of KB-R, a specific NCX inhibitor for Ca(2+) in the influx mode. Alterations in extracellular [Na] in the saline did not affect intracellular Ca(2+) transport. Caffeine, responsible for inducing Ca release from sarcoplasmic reticulum in vertebrate muscle, increased intracellular Ca(2+) compared to control, suggesting an effect of this inhibitor in gill epithelial cells of Dilocarcinus pagei, probably through release of intracellular stores. We also demonstrate here that intracellular Ca(2+) in gill cells of Dilocarcinus pagei was kept relatively constant in face of an extracellular Ca concentration of 50-fold, suggesting that crustaceans are able to display Ca(2+) homeostasis through various Ca(2+) intracellular sequestration mechanisms and/or plasma membrane Ca(2+) influx and outflux that are highly regulatory. In summary, studies using whole gill cells are an interesting approach for working with real regulatory Ca(2+) mechanisms in intact cells under physiological Ca levels (mM range), compared to earlier work using isolated vesicles of various epithelial cells. PMID- 20012438 TI - Potential role of the steroid receptor pattern in the response of inoperable intra-abdominal desmoid to toremifene after failure of tamoxifen therapy. PMID- 20012439 TI - Diagnosing lower gastrointestinal tuberculosis: colonoscopy is essential. PMID- 20012440 TI - Colonoscopy screening for individuals aged 40-49 years with a family history of stomach cancer in Korea. AB - BACKGROUND: For asymptomatic individuals aged 40-49 years, screening for colorectal cancer is recommended only if individuals are at an increased risk. A family history of colon cancer (FHCRC) is considered to increase risk, however, whether family history of stomach cancer (FHSC) increases the risk of adenoma is not well known. We aimed to see whether FHSC increases the risk in asymptomatic individuals aged 40-49 years in Korea. METHODS: Among asymptomatic individual aged 40-49 years who underwent colonoscopy screening, risk of adenoma was assessed according to FHCRC or FHSC. RESULTS: Of 3,160 individuals, male comprised 1,602 individuals (50.7%). FHCRC and FHSC was noticed in 163 (5.2%) and 227 (7.0%) individuals, respectively. Prevalence of adenoma was 31.9 %, 28.8%, and 22.3% for individuals with FHCRC, individuals with FHSC, and individuals without family history of cancer, respectively. FHSC was an independent risk factor for adenoma (odds ratio, 1.38; 95% confidence interval, 1.02-1.87, P = 0.039) in asymptomatic individuals aged 40-49 years. Compared with individuals with FHCRC, individuals with FHSC showed no difference in risk for adenoma (P = 0.347). CONCLUSIONS: As with individuals with FHCRC, individuals with FHSC might need to be considered as an individual with increased risk for adenoma. Therefore, earlier screening might be needed for asymptomatic individuals aged 40 49 years with FHSC in Korea. PMID- 20012441 TI - Association of the tumor necrosis factor-alpha -308 G/A polymorphism with nasal polyposis. AB - Nasal polyposis (NP) is a chronic inflammatory disease in which several molecular and cellular interactions play important roles. Tumor necrosis factor-alpha (TNF alpha) is a major pro-inflammatory cytokine with a key role in immune inflammatory responses in NP. Altered levels of TNF-alpha, which may occur due to polymorphisms in the TNF-alpha promoter region, may also be associated with NP susceptibility. Given these facts, we investigated the possible association of the TNF-alpha -308 G/A single nucleotide polymorphism (SNP) with NP. In this study, 97 consecutive adult patients with NP and 95 age- and gender-matched controls were recruited. For identification of SNP, restriction fragment length polymorphism analysis after polymerase chain reaction was carried out. The NP group had a significantly higher rate of polymorphism compared to controls (p = 0.015). Logistic regression analysis revealed that the presence of the TNF-alpha 308 G/A SNP is an independent risk factor for NP development (OR, 3.68; CI, 1.27 10.7; p = 0.016). The presence of a mutation failed to influence disease severity on the basis of resistance to medical and/or surgical treatment. This study suggests a possible linkage of a SNP in the TNF-alpha promoter with NP. These results need to be confirmed with multicentre studies for more precise interpretation and corroborative studies for investigating the influence of polymorphism on transcriptional activity. PMID- 20012442 TI - Analysis of 21 X-chromosomal STRs in an Algerian population sample. AB - Twenty-one X-chromosomal short tandem repeat loci, including the six clusters of linked markers DXS10148-DXS10135-DXS8378 (Xp22), DXS7132-DXS10074-DXS10079 (Xq12), DXS6801-DXS6809-DXS6789 (Xq21), DXS7424-DXS101 (Xq22), DXS10103-HPRTB DXS10101 (Xq26), DXS8377-DXS10146-DXS10134-DXS7423-DXS10011 (Xq28), and the loci DXS6800 and GATA172D05 were typed in a northwestern Algerian population sample (n = 210; 104 men and 106 women). Allele and haplotype frequencies were calculated. No evidence of linkage disequilibrium was observed between pairs of loci within clusters of linked markers. At locus DXS10148, sequence analysis of a subset of alleles displaying unusual amplicon length (>/= 36 repeat units) and anomalous electrophoretic mobility showed that this marker has a complex molecular structure with different repeat variants within alleles of identical amplicon size. PMID- 20012443 TI - Murine mCLCA5 is expressed in granular layer keratinocytes of stratified epithelia. AB - CLCA proteins represent a large family of proteins widely expressed in mammalian tissues with a unique expression pattern for each family member analyzed so far. However, their functions in normal and diseased tissues are poorly understood. Here, we present the cellular expression pattern of mCLCA5 in murine tissues using immunohistochemistry, confocal laser scanning microscopy and immune electron microscopy with specific antibodies and RT-qPCR following laser-capture microdissection. The mCLCA5 protein was localized to granular layer keratinocytes of virtually all stratified squamous epithelia of the body. Biochemical protein characterizations revealed that the amino-terminal cleavage product is fully secreted by the cell, while the carboxy-terminal cleavage product remains associated with the cell. The results imply that mCLCA5 may play a role in maturation and keratinization of squamous epithelial cells. PMID- 20012444 TI - Correlates shift work with increased risk of gastrointestinal complaints or frequency of gastritis or peptic ulcer in H. pylori-infected shift workers? AB - AIM: There is an ongoing discussion whether work under shift work conditions enhances the ulcerogenic potential of Helicobacter pylori infections. This may be induced by a disruption of the circadian rhythm. The present study assessed whether there is an association between H. pylori infection and an increased risk of gastrointestinal complaints or frequency of gastritis or peptic ulcer in H. pylori-infected shift workers. METHODS: We analyzed 615 workers divided in shift and daytime. Upper gastrointestinal diseases, medication or operation, unspecific gastrointestinal complaints, smoking habits, subjective job strain and private stress factors were monitored. The H. pylori status was analyzed using IgA and IgG antibody concentration in blood and qualitative ELISA for feces. RESULTS: It was observed that 27.6% of all workers, 34.6% of the shift workers and 16% of daytime workers were Hp positive (P = 0.004). Shift workers reported reflux or pyrosis less frequently than daytime workers. No increased lifetime prevalence for gastritis, ulcus or eradication therapy was found for shift workers. Analysis revealed a weak regression between age and Hp-positive gastritis or upper gastrointestinal complaints, smoking habits were a major confounding factor for ulcus disease. Clinical manifestation of gastritis or ulcus disease did not correlate with shift work or especially shift work including night work. CONCLUSIONS: As a result of the present study, it cannot be confirmed that there is a causal connection between shift work and upper or unspecific gastrointestinal complaints or gastritis or peptic ulcer in dependency of a H. pylori colonization. PMID- 20012445 TI - Ultra short-term heart rate recovery after maximal exercise in continuous versus intermittent endurance athletes. AB - This study tested the hypothesis that athletes participating in intermittent sports would exhibit a faster heart rate recovery (HRR) during the initial phase (<30 s) following maximal exercise than athletes participating in continuous endurance sports. Forty-six male athletes were allocated into continuous (CNT, n = 24) or intermittent groups (INT, n = 22), matched for age and aerobic fitness. Athletes performed maximal exercise on a treadmill using the ramp protocol. Immediately upon exercise cessation, subjects were placed supine with continuous measurement of HR during the first minute of recovery. Data were analyzed in 10-s intervals and compared between the groups. Repeated measures ANOVA showed a group x time interaction effects (p 0.05). The CON group showed a significant improvement in their estimation of distance completed in both the 4 km (24.6 +/- 18.2 vs. 8.2 +/- 5.5%) and 6 km (15.2 +/- 7 vs. 8.6 +/- 3%) distances (t (7) = 2.791, t (7) = 3.118, p < 0.05). No significant differences in distance estimation were observed in the EXP group (p > 0.05). In conclusion, participants in the CON group displayed a significant improvement in their judgement of distance completed, despite no improvement in completion time. This suggests that a learned pacing schema is robust and not negatively affected by subsequent pacing variation. The internal clock shows an ability to be calibrated to absolute distance, although this does not improve performance. PMID- 20012450 TI - Effect of performance level on pacing strategy during a 10-km running race. AB - The aim of this study was to examine the influence of the performance level of athletes on pacing strategy during a simulated 10-km running race, and the relationship between physiological variables and pacing strategy. Twenty-four male runners performed an incremental exercise test on a treadmill, three 6-min bouts of running at 9, 12 and 15 km h(-1), and a self-paced, 10-km running performance trial; at least 48 h separated each test. Based on 10-km running performance, subjects were divided into terziles, with the lower terzile designated the low-performing (LP) and the upper terzile designated the high performing (HP) group. For the HP group, the velocity peaked at 18.8 +/- 1.4 km h(-1) in the first 400 m and was higher than the average race velocity (P < 0.05). The velocity then decreased gradually until 2,000 m (P < 0.05), remaining constant until 9,600 m, when it increased again (P < 0.05). The LP group ran the first 400 m at a significantly lower velocity than the HP group (15.6 +/- 1.6 km h(-1); P > 0.05) and this initial velocity was not different from LP average racing velocity (14.5 +/- 0.7 km h(-1)). The velocity then decreased non significantly until 9,600 m (P > 0.05), followed by an increase at the end (P < 0.05). The peak treadmill running velocity (PV), running economy (RE), lactate threshold (LT) and net blood lactate accumulation at 15 km h(-1) were significantly correlated with the start, middle, last and average velocities during the 10-km race. These results demonstrate that high and low performance runners adopt different pacing strategies during a 10-km race. Furthermore, it appears that important determinants of the chosen pacing strategy include PV, LT and RE. PMID- 20012451 TI - Choice of surgical approach for ossification of the posterior longitudinal ligament in combination with cervical disc hernia. AB - Ossification of the posterior longitudinal ligament (OPLL) is a common spinal disorder that presents with or without cervical myelopathy. Furthermore, there is evidence suggesting that OPLL often coexists with cervical disc hernia (CDH), and that the latter is the more important compression factor. To raise the awareness of CDH in OPLL for spinal surgeons, we performed a retrospective study on 142 patients with radiologically proven OPLL who had received surgery between January 2004 and January 2008 in our hospital. Plain radiograph, three-dimensional computed tomography construction (3D CT), and magnetic resonance imaging (MRI) of the cervical spine were all performed. Twenty-six patients with obvious CDH (15 of segmental-type, nine of mixed-type, two of continuous-type) were selected via clinical and radiographic features, and intraoperative findings. By MRI, the most commonly involved level was C5/6, followed by C3/4, C4/5, and C6/7. The areas of greatest spinal cord compression were at the disc levels because of herniated cervical discs. Eight patients were decompressed via anterior cervical discectomy and fusion (ACDF), 13 patients via anterior cervical corpectomy and fusion (ACCF), and five patients via ACDF combined with posterior laminectomy and fusion. The outcomes were all favorable. In conclusion, surgeons should consider the potential for CDH when performing spinal cord decompression and deciding the surgical approach in patients presenting with OPLL. PMID- 20012452 TI - Pulsatile wall movement of spinal arachnoid cyst deteriorates spinal cord symptoms: report of three cases. AB - INTRODUCTION: Symptomatic spinal arachnoid cyst is a rare disease and the pathophysiology causing spinal cord symptoms has not been well clarified. PATIENTS AND METHODS: The authors report three symptomatic cases of spinal arachnoid cyst at the thoracic level. These patients, aged from 70 to 73 years, showed progressive gait disturbance for a few months before admission. Phase contrast cine magnetic resonance imaging demonstrated significant compression at the rostral side of the cyst during the diastolic phase of the cardiac cycle. Intraoperative ultrasonography demonstrated that the maximum expansion of the cyst and compression of the dorsal spinal cord occurred when the cerebrospinal fluid moved rostrally during diastole. All patients showed good improvement of their symptoms after surgical removal of the arachnoid cyst. CONCLUSION: This report proposes the pathophysiology that the pulsatile enlargement of the arachnoid cyst during diastolic cardiac phase can be an important factor for deterioration of spinal cord symptoms. PMID- 20012454 TI - Theoretical study on the molecular and electronic properties of some substances used for diabetes mellitus treatment. AB - Diabetes mellitus (DM) is a disease that affects a large number of people, and the number of problems associated with the disease has been increasing in the past few decades. These problems include cardiovascular disorders, blindness and the eventual need to amputate limbs. Therefore, the quality of life for people living with DM is less than it is for healthy people. In several cases, metabolic syndrome (MS), which can be considered a disturbance of the lipid metabolism, is associated with DM. In this work, two drugs used to treat DM, pioglitazone and rosiglitazone, were studied using theoretical methods, and their molecular properties were related to the biological activity of these drugs. From the results, it was possible to correlate the properties of each substance - particularly electronic properties - with the biological interactions that are linked to their pharmacological effects. These results suggest that there are future prospects for designing or developing new drugs based on the correlation between theoretical and experimental properties. PMID- 20012453 TI - Accurate DNA synthesis by Sulfolobus solfataricus DNA polymerase B1 at high temperature. AB - The accuracy of DNA synthesis by DNA polymerase B1 from the hyperthermophilic archaeon Sulfolobus solfataricus (Sso pol B1) at near the physiological temperature was investigated using M13-based mutational assays. Sso pol B1 showed replication fidelity similar to or higher than most viral, bacterial, and eukaryotic replicases. The fidelity of the enzyme was about three times as high at 70 degrees C as at 55 degrees C. Approximately two-thirds of the errors made by the enzyme were single-base substitutions, of which 58% were C --> T transition. Frameshift mutations, mostly resulting from single-base deletions, accounted for 19% of the total errors. An exonuclease-deficient mutant of Sso pol B1 was three times as mutagenic as the wild-type enzyme, suggesting that the intrinsic proofreading function contributed only modestly to the fidelity of the enzyme. Kinetic assays showed that the frequencies of all possible misincorporations by an exonuclease-deficient triple-point mutant of Sso pol B1 ranged from 5.4 x 10(-5) to 4.6 x 10(-4). The high fidelity of this enzyme in DNA synthesis was based primarily on K (m) difference rather than V (max) difference. These properties of Sso pol B1 are consistent with the proposed role of the enzyme as a replicase in S. solfataricus. PMID- 20012455 TI - Incarcerated Amyand's hernia in a premature infant associated with circumcision: a case report and literature review. AB - Amyand's hernia is a rare presentation of an appendix within an inguinal hernia sac. It is commonly mistaken for an incarcerated or strangulated hernia. Prompt diagnosis requires awareness of this entity, as well as associated radiologic findings on computed tomography (CT) and ultrasound. Treatment includes antibiotics and surgical intervention involving appendectomy and hernia repair. We present a case of a premature infant who developed systemic symptoms after a circumcision and was eventually diagnosed with an Amyand's hernia with concurrent appendicitis. PMID- 20012456 TI - Litigation following groin hernia repair in England. AB - PURPOSE: Since 1995, litigation following surgical procedures has cost the National Health Service (NHS) over 1.3 billion GBP (Great British Pounds)/2.1 billion USD (United States Dollars)/1.4 billion Euros. Despite it being the most commonly undertaken general surgical operation, no study has examined clinical negligence claims in England following groin hernia repairs. METHODS: Data from the NHS Litigation Authority of all claims made from 1995 to 2009 was obtained and interrogated. RESULTS: In total, 398 claims were made. Of these, 209 cases had been settled, of which 144 (46.6%) were in favour of the claimant to a cost of 7.35 million GBP/12 million USD/7.93 million Euros. Testicular injury and chronic pain featured in 40% of all claims. Visceral injuries and injuries requiring corrective procedures were the only predictors of a successful claim (P = 0.015 and P = 0.002, respectively). Claims associated with visceral and vascular injuries were more likely to occur in laparoscopic than in open repairs. Sexual dysfunction and chronic pain resulted in the highest average payouts of 85,467 GBP/140,565 USD/92,177 Euros and 81,288 GBP/133,693 USD/87,674 Euros, respectively. CONCLUSION: Patients should be fully informed of the incidence of testicular injury and chronic pain during the consent process. Approaches minimising visceral and vascular injury particularly in laparoscopic repair should be adopted to reduce litigation and improve patient care. PMID- 20012457 TI - The apes' edge: positional learning in chimpanzees and humans. AB - A wide variety of organisms produce actions and signals in particular temporal sequences, including the motor actions recruited during tool-mediated foraging, the arrangement of notes in the songs of birds, whales and gibbons, and the patterning of words in human speech. To accurately reproduce such events, the elements that comprise such sequences must be memorized. Both memory and artificial language learning studies have revealed at least two mechanisms for memorizing sequences, one tracking co-occurrence statistics among items in sequences (i.e., transitional probabilities) and the other one tracking the positions of items in sequences, in particular those of items in sequence-edges. The latter mechanism seems to dominate the encoding of sequences after limited exposure, and to be recruited by a wide array of grammatical phenomena. To assess whether humans differ from other species in their reliance on one mechanism over the other after limited exposure, we presented chimpanzees (Pan troglodytes) and human adults with brief exposure to six items, auditory sequences. Each sequence consisted of three distinct sound types (X, A, B), arranged according to two simple temporal rules: the A item always preceded the B item, and the sequence edges were always occupied by the X item. In line with previous results with human adults, both species primarily encoded positional information from the sequences; that is, they kept track of the items that occurred in the sequence edges. In contrast, the sensitivity to co-occurrence statistics was much weaker. Our results suggest that a mechanism to spontaneously encode positional information from sequences is present in both chimpanzees and humans and may represent the default in the absence of training and with brief exposure. As many grammatical regularities exhibit properties of this mechanism, it may be recruited by language and constrain the form that certain grammatical regularities take. PMID- 20012458 TI - Wegener's granulomatosis detected initially by integrated 18F-fluorodeoxyglucose positron emission tomography/computed tomography. AB - Early diagnosis is crucial for effective treatment of Wegener's granulomatosis, although this disease shows only atypical symptoms in the primary stage. This report describes a patient suspected of having a malignancy based on integrated 18F-fluorodeoxyglucose positron emission tomography/computed tomography (PET/CT), which showed increased uptake in pulmonary nodules and nasopharyngeal mucosa. Integrated PET/CT is therefore considered to be useful to confirm the distribution and determine the optimal site for biopsy. PMID- 20012459 TI - Mid-term results of joint-preserving procedures by a modified Mann method for big toe deformities in rheumatoid patients undergoing forefoot surgeries. AB - This study aimed to evaluate joint-preserving procedures by a modified Mann method for rheumatoid forefoot deformities and their functional outcomes in the mid-term. Eleven feet in seven patients underwent forefoot surgery using a modified Mann method for the big toe, combined with offset osteotomy or resection arthroplasty of the lesser toes. The mean follow-up period was 3.6 years. The mean score on the Japanese Society for Surgery of the Foot scale for rheumatoid arthritis foot and ankle joints improved from 44.0 to 72.0. The mean hallux valgus angle improved from 39.4 degrees to 20.5 degrees and the mean M1M5 angle improved from 31.1 degrees to 25.8 degrees . However, deformities involving a hallux valgus angle of more than 25.0 degrees recurred in three feet at the latest follow-up, although the patients did not complain of any symptoms from the recurrence. Improvement in the Sharp score for joint space narrowing was observed in the big toe, indicating better congruity of the metatarsophalangeal joint. For restraint of rheumatoid forefoot deformities, a modified Mann method, combined with offset osteotomy or resection arthroplasty, was satisfactory for not only improving the foot function, but also preserving the metatarsophalangeal joint mobility. PMID- 20012460 TI - Antioxidant potential, paraoxonase 1, ceruloplasmin activity and C-reactive protein concentration in diabetic retinopathy. AB - The aim of this study was to evaluate the ferric-reducing ability of serum (FRAS), paraoxonase 1 (PON1), ceruloplasmin serum oxidase activity and hsCRP level in patients with type1 diabetes mellitus without and with diabetic retinopathy. The study was performed in 76 patients with type 1 diabetes mellitus, 35 without diabetic retinopathy (group 1) and 41 with preproliferative and proliferative retinopathy (group 2). Control group consisted of 35 nondiabetic, age-, gender-, body mass-matched healthy volunteers who came to the outpatient clinic for a routine health check-up. We evaluated FRAS using the method described by Benzie and Strain; PON1 by kinetic spectrophotometric assay with paraoxon as substrate and ceruloplasmin using its oxidative activity with 3 phenylenodiamine as substrate. CRP was measured with a high sensitive enzyme immunoassay. PON1 activity was significantly decreased in patients with diabetic retinopathy (227.66 +/- 123.57 U/l) when compared with control (312.04 +/- 129.77 U/l). FRAS was significantly decreased in group 2 (439.33 +/- 79.87 micromol/l) when compared with group 1 (522.79 +/- 167.56 micromol/l) and control (529.80 +/- 81.99 micromol/l). Ceruloplasmin activity was significantly elevated in group 1 (58.36 +/- 22.56 U/g protein) when compared with control (45.22 +/- 14.96 U/g protein). We have found significant increase in hsCRP level in group 2 (3.71 +/- 2.47 mg/l) when compared with group 1 (1.75 +/- 1.01 mg/l) and control (0.57 +/- 0.46 mg/l). The PON1/CRP ratio in control group was significantly increased when compared with diabetic patients and was significantly decreased in group 2 compared with group 1. We have not found gender-dependent difference in studied parameters in both control and in study groups. We have found tendency to decrease the serum activity of FRAS and hsCRP in elder patients but the difference was significant only in group 2. FRAS and PON 1 activity is decreased in patients with type 1 diabetes mellitus with presence of diabetic retinopathy which confirms that oxidative stress could play a role in pathogenesis of diabetic retinopathy. Significantly elevated levels of hsCRP in diabetic patients with the presence of diabetic retinopathy compared with patients without diabetic retinopathy providing a link between inflammation and the development of microvascular complication of diabetes. Because of the significant difference in PON1/CRP ratio between patients without and with the presence of diabetic retinopathy, it seems that PON1:CRP ratio may be used as a biochemical marker for progression of retinopathy. The link between the antioxidant concentration, inflammation and the development of diabetes complications needs further longitudinal studies in order to confirm our findings. PMID- 20012462 TI - Decrease in expression of histamine H2 receptors by human amniotic epithelial cells during differentiation into pancreatic beta-like cells. AB - OBJECTIVE AND DESIGN: Considering the role of histaminergic pathway in the differentiation of stem cells, we compared expression patterns of H(1) and H(2) receptors in the human amniotic epithelial cells (HAEC) culture at different stages of nicotinamide-induced differentiation into PBLC with the control HAEC. MATERIAL AND METHODS: HAEC isolated after term pregnancies (N = 12) were cultured in vitro. Altogether, 72 cultures were established. The culture medium in the studied group was supplemented on Day 5 with nicotinamide (10 mM). C-peptide concentration in the medium collected every 3 days for 15 days was determined immunoenzymatically as a marker of differentiation. At the same intervals the cultures were formalin-fixed and paraffinembedded for H(1) and H(2) receptors immunostaining. Quantitative immunohistochemistry was applied for evaluation of H(1) and H(2) expression. RESULTS: C-peptide was detected on Day 6 and the levels were kept gradually increased until Day 12, then stayed at almost the same level, 3.7-fold higher than initially. Expression of H(2) was unchanged until Day 9 after nicotinamide addition, then was significantly (p < 0.05) decreased and amounted (mean % value for the measurements performed on Day 12 and Day 15, +/ SEM) 49.73 +/- 11.03 of the reference value obtained in control HAEC. CONCLUSION: Variable expression of H(2) during nicotinamide-induced differentiation of HAEC into PBLC may define a time-point, indicating involvement of histamine at the earlier stages. PMID- 20012461 TI - The alpha-kinase family: an exceptional branch on the protein kinase tree. AB - The alpha-kinase family represents a class of atypical protein kinases that display little sequence similarity to conventional protein kinases. Early studies on myosin heavy chain kinases in Dictyostelium discoideum revealed their unusual propensity to phosphorylate serine and threonine residues in the context of an alpha-helix. Although recent studies show that some members of this family can also phosphorylate residues in non-helical regions, the name alpha-kinase has remained. During evolution, the alpha-kinase domains combined with many different functional subdomains such as von Willebrand factor-like motifs (vWKa) and even cation channels (TRPM6 and TRPM7). As a result, these kinases are implicated in a large variety of cellular processes such as protein translation, Mg(2+) homeostasis, intracellular transport, cell migration, adhesion, and proliferation. Here, we review the current state of knowledge on different members of this kinase family and discuss the potential use of alpha-kinases as drug targets in diseases such as cancer. PMID- 20012465 TI - A well deserved honor: Yvonne Martin, 2009 recipient of the Herman Skolnik Award. PMID- 20012464 TI - Efficacy of intracerebral delivery of cisplatin in combination with photon irradiation for treatment of brain tumors. AB - We have evaluated the efficacy of intracerebral (i.c.) convection-enhanced delivery (CED) of cisplatin in combination with photon irradiation for the treatment of F98 glioma-bearing rats. One thousand glioma cells were stereotactically implanted into the brains of Fischer rats and 13 days later cisplatin (6 microg/20 microl) was administered i.c. by CED at a flow rate of 0.5 microl/min. On the following day the animals were irradiated with a single 15 Gy dose of X-rays, administered by a linear accelerator (LINAC) or 78.8 keV synchrotron X-rays at the European Synchrotron Radiation Facility (ESRF). Untreated controls had a mean survival time (MST) + or - standard error of 24 + or - 1 days compared to >59 + or - 13 days for rats that received cisplatin alone with 13% of the latter surviving >200 days. Rats that received cisplatin in combination with either 6 MV (LINAC) or 78.8 keV (synchrotron) X-rays had almost identical MSTs of >75 + or - 18 and >74 + or - 19 days, respectively with 17 and 18% long-term survivors. Microscopic examination of the brains of long-term surviving rats revealed an absence of viable tumor cells and cystic areas at the presumptive site of the tumor. Our data demonstrate that i.c. CED of cisplatin in combination with external X-irradiation significantly enhanced the survival of F98 glioma-bearing rats. This was independent of the X-ray beam energy and probably was not due to the production of Auger electrons as we previously had postulated. Our data provide strong support for the approach of concomitantly administering platinum-based chemotherapy in combination with radiotherapy for the treatment of brain tumors. Since a conventional LINAC can be used as the radiation source, this should significantly broaden the clinical applicability of this approach compared to synchrotron radiotherapy, which could only be carried out at a very small number of specialized facilities. PMID- 20012467 TI - Long-term metabolic effects of different doses of niacin-bound chromium on Sprague-Dawley rats. AB - We simultaneously assessed benefits and risks of niacin-bound chromium (NBC) intake at varying doses over a prolonged period of time (>1.2 years) in male and female Sprague-Dawley (SD) rats. We performed the study in two phases. First, we followed 60 male and 60 female SD rats, each gender divided into six groups. Through day 150 (phase 1A), all SD rats received a high sucrose diet (30% w/w) with or without different concentrations of NBC. The male/female groups were: 1] control without NBC n = 10, 2] low NBC (2.8 ppm, n = 10), 3] medium NBC (8.7 ppm, n = 20), 4] high NBC (28.0 ppm, n = 20). Based on dosing, we refer to the three treatment groups as 1X, 3X, and 10X. During days 151-312 (phase 1B), NBC was removed from diets of one half of the 3X and 10X groups. These are referred to as 3X satellite and 10X satellite. In phase 2 (days 313-460), males from groups 1X, 3X, 10X, 3X satellite, and 10X satellite received the same 3X dose of NBC (8.7 ppm). The last two groups also ingested different doses of a formulation of natural products in addition to NBC. We examined blood pressure, the renin angiotensin system (RAS), nitric oxide (NO), and insulin systems and inflammatory parameters. Results in male and female SD rats were comparable. NBC lowered systolic blood pressure (SBP) in a dose-dependent fashion; however, after 200 days, the SBP of the low dose group (1X) began to rise and returned to baseline control. After raising the dose of NBC to 3X, the SBP in the 1X group decreased significantly once more. When half the test rats (3X and 10X) were deprived of NBC, SBP rose gradually to control levels after 2 to 3 months. However, the SBP decreased significantly once more when each satellite group returned to the 3X dose. Special testing suggests that NBC at adequate dosing increases insulin sensitivity, lowers HbA1C, decreases activity of the RAS, at least in part, through ACE inhibition, enhances NO activity, and is without signs of toxicity. The addition of a formula composed of antioxidants and immune modulators to the chromium regimen caused even faster and more profound changes in SBP than with NBC alone. We conclude that NBC at adequate dosing is effective in male and female SD rats on certain metabolic parameters over a prolonged period, effects that disappear over months after NBC is removed. When dosing is returned, the effectiveness of NBC returns. Low doses of NBC may lose their effect over time. No signs of toxicity were observed. PMID- 20012466 TI - Genomic distribution of retrotransposons 297, 1731, copia, mdg1 and roo in the Drosophila melanogaster species subgroup. AB - The intragenomic distribution of five retrotransposon families (297, 1731, copia, mdg1 and roo) in the species of the melanogaster complex was studied by comparing results of the Southern blotting technique in males and females with those of in situ hybridization. The degree of structural polymorphism of each family in the different species was also investigated by restriction enzyme analysis. It was found that genomic distribution is a trait that depends on the family and species. The distribution of roo is mainly euchromatic in the four species and 1731 is heterochromatic, but the distribution of families 297, copia and mdg1 is markedly different in the melanogaster and simulans clades. These families were mainly euchromatic in D. melanogaster but heterochromatic in its sibling species. In the simulans clade most copia and mdg1 elements are located on chromosome Y. Differences in genomic distribution are unrelated with structural conservation. The relation of intragenomic distribution to phylogeny, transpositional activity and the role of the host genome are discussed. PMID- 20012468 TI - Transduction systems - guanylate cyclase. Preface. PMID- 20012471 TI - The impacts of gaming expansion on economic growth: a theoretical reconsideration. AB - This paper employs a general equilibrium framework to analyze the effects on economic growth of global expansions in casino gaming, which exports gambling services largely to non-residents. Both domestic and foreign investments in the gaming sector bring in not only substantial revenues but also positive spillover effects on related sectors and even on the entire local economy. However, an over expansion of commercial gambling may lead to deterioration in the terms of trade with an adverse impact on real income. If this situation persists, it would not be impossible for immiserizing growth to occur. As a highly profitable sector, casino gaming may enable its operators to diversify out of this risk if they invest retained profits in non-gaming sectors to cash in on the spillover effects it has created. The gaming-dominant economy can then be directed on a more balanced and sustainable growth path, and will become less susceptible to business cycles. Indeed, economic experiences in the world's major casino resorts are consistent basically with this argument for diversification. We believe that after the current global crisis fades away, economic growth and resulting surges in global demand for gambling services can provide further opportunities for the expansion of existing casino resorts and the development of new gaming markets. PMID- 20012470 TI - FOXO transcription factors and VEGF neutralizing antibody enhance antiangiogenic effects of resveratrol. AB - Resveratrol (trans-3,5,4'-trihydroxystilbene), a compound found largely in the skins of red grapes and wines, possesses anti-cancer and anti-angiogenic properties and protects the cardiovascular system. However, the molecular mechanisms by which resveratrol inhibits angiogenesis are currently subjects of intense investigation. The purpose of this study was to examine whether FOXO transcription factors mediate anti-angiogenic effects of resveratrol, and whether vascular endothelial growth factor (VEGF) neutralizing antibody can enhance these effects of resveratrol. Inhibition of PI3 kinase (PI3K)/AKT and MEK/ERK pathways synergistically inhibited migration and capillary tube formation of Human Umbilical Vein Endothelial Cells (HUVECs) and further enhanced the anti angiogenic effects of resveratrol. Inhibitors of AKT and MEK kinase synergistically inhibited cytoplasmic FOXO3a phosphorylation, which was accompanied by its nuclear translocation in HUVECs. Interestingly, inhibition of PI3K/AKT and MEK/ERK pathways synergistically induced FOXO transcriptional activity and inhibited cell migration and capillary tube formation. Antiangiogenic effects of resveratrol were enhanced by inhibitors of AKT and MEK. Phosphorylation-deficient mutants of FOXOs induced FOXO transcriptional activity, inhibited HUVEC cell migration, and capillary tube formation, and also enhanced antiangiogenic effects of resveratrol. Finally, VEGF neutralizing antibody enhanced the anti-proliferative and anti-angiogenic effects of resveratrol. In conclusion, regulation of FOXO transcription factors by resveratrol may play an important role in angiogenesis which is critical for cancer, diabetic retinopathy, rheumatoid arthritis, psoriasis, and cardiovascular disorders. PMID- 20012472 TI - Moral agency, identity crisis and mental health: an anthropologist's plight and his Hmong ritual healing. AB - During anthropological fieldwork, the author had a serious accident on the outskirts of a Hmong village in the highland of Laos. However, this dramatic incident turned out to be the occasion of his ritual initiation into the local village community. An analysis of narratives of the incident reveals Hmong conceptions of the anthropologist's physical, mental and moral affliction, its causative concomitants and his ritual healing. Hmong mental health and identity are situated in a moral space of exchange relationships to significant others, challenging basic assumptions of concepts of the person widely held in psychiatry and beyond. The healing ritual transformed the author's being from indeterminate "other," in a life-threatening state of identity crisis, to a wholesome Hmong "self," in a state of health and moral agency. This exemplary rite de passage highlights the affinity of ritual healing and constitution of self in a moral space. The underlying relational concept of the person is in sharp contrast to psychiatry's concepts of the person, which are deeply shaped by values of individualism. Psychiatric services must accommodate substantial differences in the concepts of the person when treating Hmong migrants from Laos. PMID- 20012469 TI - New insight into the functioning of nitric oxide-receptive guanylyl cyclase: physiological and pharmacological implications. AB - The cellular counterpart of the "soluble" guanylyl cyclase found in tissue homogenates over 30 years ago is now recognized as the physiological receptor for nitric oxide (NO). The ligand-binding site is a prosthetic haem group that, when occupied by NO, induces a conformational change in the protein that propagates to the catalytic site, triggering conversion of GTP into cGMP. This review focuses on recent research that takes this basic information forward to the beginnings of a quantitative depiction of NO signal transduction, analogous to that achieved for other major transmitters. At its foundation is an explicit enzyme-linked receptor mechanism for NO-activated guanylyl cyclase that replicates all its main properties. In cells, NO signal transduction is subject to additional, activity dependent modifications, notably through receptor desensitization and changes in the activity of cGMP-hydrolyzing phosphodiesterases. The measurement of these parameters under varying conditions in rat platelets has made it possible to formulate a cellular model of NO-cGMP signaling. The model helps explain cellular responses to NO and their modification by therapeutic agents acting on the guanylyl cyclase or phosphodiesterase limbs of the pathway. PMID- 20012473 TI - Perceptions of school nurses on the challenges of service provision to ESL students. AB - The number of English-as-as second-language (ESL) students in public schools across the country is rising creating unique challenges for school health services with the introduction of new cultures and languages. This study describes the perceptions of school nurses in Alabama schools regarding the challenges they face when communicating with ESL students and their parents. The population consists of school nurses in Alabama public schools. The obtained sample of 1,429 responses is presented and discussed. Urban school nurses were more likely to have access to an interpreter, yet they were more likely to rely on an ESL student to act as a translator when speaking to his/her parents. Kindergarten through elementary school nurses were more likely to witness an increase in ESL students and encounter difficulty communicating with these students and their parents. School nurses have a unique opportunity to engage parents of ESL students in the health of their child, thus it is vital that they be provided with cultural facts and encouraged to use interpreter services to lessen language barriers. PMID- 20012474 TI - Leveraging geospatial data, technology, and methods for improving the health of communities: priorities and strategies from an expert panel convened by the CDC. AB - In 2008, CDC convened an expert panel to gather input on the use of geospatial science in surveillance, research and program activities focused on CDC's Healthy Communities Goal. The panel suggested six priorities: spatially enable and strengthen public health surveillance infrastructure; develop metrics for geospatial categorization of community health and health inequity; evaluate the feasibility and validity of standard metrics of community health and health inequities; support and develop GIScience and geospatial analysis; provide geospatial capacity building, training and education; and, engage non-traditional partners. Following the meeting, the strategies and action items suggested by the expert panel were reviewed by a CDC subcommittee to determine priorities relative to ongoing CDC geospatial activities, recognizing that many activities may need to occur either in parallel, or occur multiple times across phases. Phase A of the action items centers on developing leadership support. Phase B focuses on developing internal and external capacity in both physical (e.g., software and hardware) and intellectual infrastructure. Phase C of the action items plan concerns the development and integration of geospatial methods. In summary, the panel members provided critical input to the development of CDC's strategic thinking on integrating geospatial methods and research issues across program efforts in support of its Healthy Communities Goal. PMID- 20012475 TI - Recruitment, training outcomes, retention, and performance of community health advisors in two tobacco control interventions for Latinos. AB - Community Health Advisors (CHAs) are indigenous lay health advisors who, with training, can create health awareness, disseminate health information and support behavior change in their communities. Little data are available that describe the characteristics, recruitment, training, retention, and performance of CHAs. The present study described the characteristics, recruitment process, training outcomes, retention activities, and performance of two sets of CHAs who delivered tobacco-related interventions in the local Latino community. The Tobacco Control in Latino Communities (TCLC) Center trained 35 CHAs to conduct either a smoking cessation program for Spanish-speaking adult smokers or a behavioral problem solving intervention to reduce environmental tobacco smoke (ETS) exposure among low-income Latino children. Theoretical psychosocial constructs related to behavior change, general self-esteem, general self-efficacy, and demographics were collected from CHAs before and after training. Additional measures captured the level of professionalism exercised and effort undertaken by the CHAs, as well actual outcomes of their efforts. Of the 33 women and 2 men CHAs recruited, 86% were originally from Mexico, most had a high school education, most were married, and the average monthly household income was $1,100-$1,400. Mean participant age was 42 years, and level of acculturation was relatively low. There were changes in the desired direction pre-to-post training for both ETS and smoking cessation program CHAs for most of the psychosocial constructs. Expert ratings of CHA performance were good, and recipients of the CHAs' efforts showed positive changes in behavior. This information may aid in planning for recruitment and evaluation of CHAs for future tobacco control programs. PMID- 20012476 TI - Unsafe sex among HIV positive individuals: cross-sectional and prospective predictors. AB - A follow-up study was conducted on a sample of 120 ethnically diverse HIV positive men and women first interviewed in 2000. Of the 86 survivors, 37 (43%) were able to be contacted 7-8 years later to conduct an exploratory examination of cross-sectional and prospective predictors of unsafe sexual behavior. Predictors that emerged as significant in the two cross-sectional analyses and the prospective analysis tended to be different variables, perhaps underscoring changing needs, perceptions, and behaviors among HIV positive persons over time. The cross-sectional analysis conducted at the baseline time frame showed a considerable number of significant correlates of unsafe sex, including several demographic/background variables. The cross-sectional analysis conducted on data collected 7 years later, on the other hand, showed far fewer significant correlates of unsafe sex, none of which were demographic/background variables, and which tended to be different correlates than those found in the baseline cross-sectional analysis. Significant predictors in the prospective analysis tended to be social support factors. This different pattern of prediction may be important to those designing interventions to influence risky sexual behavior. PMID- 20012477 TI - Bone health in immigrant Hispanic women living in Texas. AB - Osteoporosis is a serious national public health problem, and is expected to increase significantly over the next few decades, especially in women. A limitation of bone health research exists since few studies have involved Hispanic women, and even fewer, Hispanic immigrant women. For this study we examined the effects of anthropometric, behavioral, and health history variables on bone mineral density (BMD) in 84 immigrant Hispanic women, age 40 and above. BMD was assessed at the spine, femur, and forearm using dual energy x-ray absorptiometry (DXA). Demographic information, health histories, and behavioral risk factors were obtained from a questionnaire. In the younger group (mean age = 44.1 years) 61% had spinal osteopenia, and in the postmenopausal group (mean age = 53.0 years) 59% had osteopenia and 13% had osteoporosis. Femur sites were free of osteoporosis. Mean body mass index (BMI) was 31.8 +/- 6.1 and mean waist girth was 95.6 +/- 12.5 cm, indicating overall and abdominal obesity. Partial correlations indicated a significant positive relationship between body fat variables and total femur BMD values. ANOVAs revealed no differences in BMD values at any bone site across tertile levels for calcium intake or for physical activity. However, supplemental and dietary calcium intakes were very low and few participants engaged in regular physical activity outside of work and activities of daily living (ADL). In light of the expected increase in osteoporosis in this population and the prevalence of spinal osteopenia in the younger participants, education about the health risks of osteoporosis should be made available to this group. PMID- 20012479 TI - The association of HIV counseling and testing with HIV risk behaviors in a random population-based survey in Kisumu, Kenya. AB - HIV testing has been promoted as a key HIV prevention strategy in low-resource settings, despite studies showing variable impact on risk behavior. We sought to examine rates of HIV testing and the association between testing and sexual risk behaviors in Kisumu, Kenya. Participants were interviewed about HIV testing and sexual risk behaviors. They then underwent HIV serologic testing. We found that 47% of women and 36% of men reported prior testing. Two-thirds of participants who tested HIV-positive in this study reported no prior HIV test. Women who had undergone recent testing were less likely to report high-risk behaviors than women who had never been tested; this was not seen among men. Although rates of HIV testing were higher than seen in previous studies, the majority of HIV infected people were unaware of their status. Efforts should be made to increase HIV testing among this population. PMID- 20012478 TI - How is the highly positive endocochlear potential formed? The specific architecture of the stria vascularis and the roles of the ion-transport apparatus. AB - Cochlear endolymph, an extracellular solution containing 150 mM K(+), exhibits a positive potential of +80 mV. This is called the endocochlear potential (EP) and is essential for audition. The mechanism responsible for formation of the EP has been an enigma for the half century since its first measurement. A key element is the stria vascularis, which displays a characteristic tissue structure and expresses multiple ion-transport apparatus. The stria comprises two epithelial layers: a layer of marginal cells and one composed of intermediate and basal cells. Between the two layers lies an extracellular space termed the intrastrial space (IS), which is thus surrounded by the apical membranes of intermediate cells and the basolateral membranes of marginal cells. The fluid in the IS exhibits a low concentration of K(+) and a positive potential similar to the EP. We have demonstrated that the IS is electrically isolated from the neighboring extracellular fluids, perilymph, and endolymph, which allows the IS to sustain its positive potential. This IS potential is generated by K(+) diffusion across the apical membranes of intermediate cells, where inwardly rectifying Kir4.1 channels are localized. The low K(+) concentration in the IS, which is mandatory for the large K(+)-diffusion potential, is maintained by Na(+),K(+)-ATPases and Na(+),K(+),2Cl(-)-cotransporters expressed at the basolateral membranes of marginal cells. An additional K(+)-diffusion potential formed by KCNQ1/KCNE1-K(+) channels at the apical membranes of marginal cells also contributes to the EP. Therefore, the EP depends on an electrically isolated space and two K(+) diffusion potentials in the stria vascularis. PMID- 20012481 TI - Staff turnover in statewide implementation of ACT: relationship with ACT fidelity and other team characteristics. AB - Staff turnover on assertive community treatment (ACT) teams is a poorly understood phenomenon. This study examined annual turnover and fidelity data collected in a statewide implementation of ACT over a 5-year period. Mean annual staff turnover across all observations was 30.0%. Turnover was negatively correlated with overall fidelity at Year 1 and 3. The team approach fidelity item was negatively correlated with staff turnover at Year 3. For 13 teams with 3 years of follow-up data, turnover rates did not change over time. Most ACT staff turnover rates were comparable or better than other turnover rates reported in the mental health and substance abuse literature. PMID- 20012483 TI - Broiler performance on finisher diets containing different levels of either Pleurotus ostreatus-fermented dried cocoa pod husk or dried cocoa pod husk supplemented with enzymes. AB - The purpose was to investigate the feeding value of fibrous cocoa pod husk (CPH) either fermented with Pleurotus ostreatus or supplemented with fibre-degrading multi-enzymes (Viscozyme L + Pectinex 5XL) before inclusion in diets for 3-week old finishing broilers. Growth performance and carcass yield of broilers fed diets containing 100, 200 or 300 g kg(-1) of the P. ostreatus-fermented CPH or 100, 200 or 300 g kg(-1) of the enzyme-supplemented CPH as well as a normal broiler finisher diet (control), were investigated in a 35-day feeding trial setup as a completely randomised design, with the seven treatments replicated three times. There were significant differences (P < 0.05) among treatments for some of the parameters measured including growth-related indices, percent weights of abdominal fat, intestine, and gizzard. It was concluded that P. ostreatus fermented dried CPH up to a level of 200 g kg(-1) and enzyme-supplemented dried CPH up to a level of 100 g kg(-1) could be included in broiler finisher diets without any deleterious effects. PMID- 20012482 TI - Anti-angiogenic tyrosine kinase inhibitors: what is their mechanism of action? AB - Tyrosine kinases are important cellular signaling proteins that have a variety of biological activities including cell proliferation and migration. Multiple kinases are involved in angiogenesis, including receptor tyrosine kinases such as the vascular endothelial growth factor receptor. Inhibition of angiogenic tyrosine kinases has been developed as a systemic treatment strategy for cancer. Three anti-angiogenic tyrosine kinase inhibitors (TKIs), sunitinib, sorafenib and pazopanib, with differential binding capacities to angiogenic kinases were recently approved for treatment of patients with advanced cancer (renal cell cancer, gastro-intestinal stromal tumors, and hepatocellular cancer). Many other anti-angiogenic TKIs are being studied in phase I-III clinical trials. In addition to their beneficial anti-tumor activity, clinical resistance and toxicities have also been observed with these agents. In this manuscript, we will give an overview of the design and development of anti-angiogenic TKIs. We describe their molecular structure and classification, their mechanism of action, and their inhibitory activity against specific kinase signaling pathways. In addition, we provide insight into what extent selective targeting of angiogenic kinases by TKIs may contribute to the clinically observed anti-tumor activity, resistance, and toxicity. We feel that it is of crucial importance to increase our understanding of the clinical mechanism of action of anti-angiogenic TKIs in order to further optimize their clinical efficacy. PMID- 20012484 TI - Efficacy and safety of lubiprostone in patients with chronic constipation. AB - AIMS: The aim of this study is to assess the efficacy and safety of lubiprostone in adults with chronic constipation. METHODS: This multicenter, parallel-group trial enrolled 237 patients with chronic constipation and randomized them to 4 weeks of double-blind treatment with oral lubiprostone 24 mcg or placebo twice daily. The primary efficacy endpoint was the number of spontaneous bowel movements (SBMs) after 1 week of treatment. Secondary evaluations included SBMs at weeks 2, 3, and 4; percentage of patients with a SBM within 24 h of first study dose; stool consistency; degree of straining; constipation severity; abdominal bloating and discomfort; global treatment effectiveness; and safety assessments. RESULTS: Lubiprostone-treated patients experienced greater mean numbers of SBMs at week 1 compared with placebo (5.89 versus 3.99, P = 0.0001), with significantly greater percentages having SBMs within 24 h of the first dose (61.3% versus 31.4%, P < 0.0001). At each assessment, SBM frequency and percentages of full responders (> or =4 SBM per week) were significantly greater among lubiprostone-treated patients compared with placebo (P < or = 0.0171). Lubiprostone-treated patients reported significant improvements in stool consistency, straining, and constipation severity at all weeks, and in abdominal bloating at week 1. Patient assessments of treatment effectiveness were significantly greater with lubiprostone compared with placebo at all weeks (P < 0.0004). Gastrointestinal-related disorders were the most common adverse events in both treatment groups. CONCLUSIONS: In patients with chronic constipation, lubiprostone produced a bowel movement in the majority of individuals within 24 h of initial dosing, with sustained improvement in frequency as well as other constipation symptoms over 4 weeks of treatment. PMID- 20012485 TI - Familial clustering and genetic background of primary biliary cirrhosis in Japan. AB - BACKGROUND: Primary biliary cirrhosis (PBC) is regarded as an autoimmune liver disease and familial clustering of PBC could represent some genetic predisposition to the disease. AIMS: To elucidate the genetic background of PBC by investigating familial cases of PBC. METHODS: Familial cases were picked out from 171 PBC patients who enrolled in this study. We analyzed them and their family members, and compared them clinically and immunogenetically to non familial cases. RESULTS: Out of 171 PBC patients, ten (5.8%) were identified as familial PBC in five families. The clinical features of familial PBC were almost comparable to those of non-familial PBC. The distribution of human leukocyte antigens (HLA)-A, -B and -DR in familial PBC showed no specificity. Two new PBC patients were identified in one family in addition to the two originally enrolled PBC patients, resulting in four patients with PBC within the same family. The two new PBC patients had an identical HLA haplotype. On the other hand, one HLA identical sister of a PBC patient in another family did not develop PBC. CONCLUSIONS: Primary biliary cirrhosis can exhibit familial clustering without any HLA predisposition, however, a survey of families for PBC could be useful for identifying new patients with PBC in the asymptomatic stage for earlier diagnosis and treatment. PMID- 20012486 TI - Panic. PMID- 20012487 TI - Inner night and inner light: a Quaker model of pastoral care for the mentally ill. AB - The same theological principles that motivated Quakers in institutional reform work continue to influence uniquely Quaker approaches to pastoral care for the mentally ill today. This unity of psychological and spiritual care, inspired by George Fox, was first apparent in the work of the Religious Society of Friends asylum reforms in the nineteenth century. These principles matured during the early twentieth century as they entered into dialogue with Jung and Jungian psychology and continue to inspire Quaker pastoral care models today. This paper will examine how theological concepts affect the way Friends approach mental health care, historically and in contemporary times. PMID- 20012488 TI - Activation of cloned BK(Ca) channels in nitric oxide-induced apoptosis of HEK293 cells. AB - The large conductance Ca(2+)-activated K(+) (BK(Ca)) channels are highly expressed in vascular smooth muscle cells (VSMCs) and play an essential role in the regulation of various physiological functions. Besides its electrophysiological function in vascular relaxation, BK(Ca) has also been reported to be implicated in nitric oxide (NO)-induced apoptosis of VSMCs. However, the molecular mechanism is not clear and has not been determined on cloned channels. The present study was designed to clarify whether activation of cloned BK(Ca) channel was involved in NO-induced apoptosis in human embryonic kidney 293 (HEK293) cell. The cDNA encoding the alpha-subunit of BK(Ca) channel, hSloalpha, was transiently transfected into HEK293 cells. The apoptotic death in HEK-hSloalpha cells was detected using immunocytochemistry, analysis of fragmented DNA by agarose gel electrophoresis, MTT test, and flow cytometry assays. Whole-cell and single-channel characteristics of HEK-hSloalpha cells exhibited functional features similar to native BK(Ca) channel in VSMCs. Exposuring of HEK- hSloalpha cells to S-nitroso-N-acetyl-penicillamine increased the hSloalpha channel activities of whole-cell and single-channel, and then increased percentage of cells undergoing apoptosis. However, blocking hSloalpha channels with 1 mM tetraethylammonia or 100 nM iberiotoxin significantly decreased the NO-induced apoptosis, whereas 30 microM NS1619, the specific agonist of BK(Ca), independently increased hSloalpha currents and induced apoptosis. These results indicated that activation of cloned BK(Ca) channel was involved in NO-induced apoptosis of HEK293 cells. PMID- 20012490 TI - Effects of extracellular calcium on the volley activity of entorhinal cortex neurons in neonatal rats: computer simulation. AB - The calcium-dependent volley activity of neurons in the entorhinal cortex of neonatal rats increased when the extracellular Ca(2+) concentration decreased from 2 to 1 mM. This may be associated with changes in the properties of the slow sodium current. Computer simulation of volley neurons using a model including a relationship between the slow sodium current and extracellular Ca(2+) supported this suggestion. PMID- 20012489 TI - Paradoxical sleep as a tool for understanding the hippocampal mechanisms of contextual memory. AB - Existing data on the involvement of the hippocampus in contextual memory and the fact that contextual memory is impaired in dreams occurring during paradoxical sleep allowed us to suggest that one of the causes of this impairment consists of changes in the efficiency of synaptic transmission in the hippocampus due to increases (as compared with waking) in the concentrations of acetylcholine, dopamine, and cortisol, as well as the absence of serotonin and noradrenaline. Our previous analysis showed that in paradoxical sleep, long-term depression can be induced all components of the polysynaptic pathway through the hippocampal formation, while potentiation can occur at the inputs from the entorhinal cortex to hippocampal fields CA1 and CA3 and in the associative connections in field CA3. It is hypothesized that the correct functioning of episodic memory requires efficient transmission of signals in each component of the polysynaptic pathway through the hippocampus, allowing a neuronal representation of the context to be created within it. In the state of waking, reproduction of the context of an episode simultaneously activates the neuronal representation of the context remembered in the hippocampus and neuronal representations of the details of the episode remembered in those areas of the cortex in which they were processed. It follows from the proposed mechanism that any neurotransmitter or neuropeptide able to promote longterm potentiation in all components of the polysynaptic pathway through the hippocampus can improve episodic memory. As the consequences of the mechanism are consistent with experimental data, it can be used to seek agents improving episodic memory. PMID- 20012491 TI - Reciprocal relationships between the oscillatory systems of the brain. AB - Resting EEG recordings were made from cohorts of 146 children aged 7-17 years and 132 adults aged 18-32 years and the levels of personality features and psychopathology were assessed using the Eysenck, Spilberger, Gray-Wilson, and Goodman questionnaires. Factor analysis was used to discriminate covariance of measures of the spectral power of EEG rhythms into positive and negative components. The latter were interpreted as a measure of inhibitory interactions between oscillatory systems. In children, positive covariance of rhythms was stronger than in adults, while reciprocal relationships between oscillatory systems were weaker. In adults, trait anxiety correlated positively with the strength of the reciprocal relationship between the alpha and delta oscillatory systems. In children, an analogous relationship was seen between anxiety and the strength of the reciprocal relationship between the theta and delta systems. The data are discussed in the light of the evolutionary interpretation of EEG rhythms. PMID- 20012492 TI - Effects of activation and blockade of dopamine receptors on the extinction of a passive avoidance reaction in mice with a depressive-like state. AB - Learning and extinction of a conditioned passive avoidance reaction resulting from neuropharmacological actions on dopamine D(1) and D(2) receptors were demonstrated to be specific in intact mice and in mice with a depressive-like state. Learning was degraded only after administration of the D(2) receptor antagonist sulpiride and was independent of the initial functional state of the mice. In intact mice, activation of D(2) receptors with quinpirole led to a deficit of extinction, consisting of a reduction in the ability to acquire new inhibitory learning in conditions associated with the disappearance of the expected punishment. In mice with the "behavioral despair" reaction, characterized by delayed extinction, activation of D(1) receptors with SKF38393 normalized this process, while the D(2) agonist was ineffective. A positive effect consisting of accelerated extinction of the memory of fear of the dark ("dangerous") sector of the experimental chamber was also seen on blockade of both types of dopamine receptor. PMID- 20012493 TI - Effects of blockade of dopamine D2 receptors on extracellular citrulline levels in the nucleus accumbens during performance of a conditioned reflex fear response. AB - Vital microdialysis studies on Sprague-Dawley rats using HPLC showed that performance of a conditioned reflex fear response was accompanied by an increase in the extracellular level of citrulline (a coproduct of nitric oxide synthesis) in the nucleus accumbens. Administration of the dopamine D(2) receptor antagonist raclopride (10 microM) into the nucleus accumbens decreased the magnitude of the increase in the extracellular citrulline level in this structure during performance of the conditioned reflex fear response but had no effect on its behavioral measures (the level of freezing). Doses increased investigative activity in a novel context which had been inhibited by acquisition of the conditioned reflex fear response, without affecting the investigative behavior of control animals. These data suggest that the dopaminergic input and dopamine D(2) receptors control the activity of the NO-ergic system of the nucleus accumbens during performance of the conditioned reflex fear response and may control "transfer" of fear to another behavioral situation. PMID- 20012494 TI - Morphometric characteristics of the brain in children aged one year (magnetic resonance tomography data). AB - The aim of the present work was to identify complex morphometric characteristics of the living brain in children aged one year with assessment of individual variation (sexual, interhemisphere) by magnetic resonance tomography. The results demonstrated sexual dimorphism in brain sizes: endbrain sizes were generally larger in boys, while structures in the stem part of the brain were larger in girls. Interhemisphere asymmetry of the brain was found in one-year-old children in most cases, lobe sizes were greater in the right hemisphere as compared with the left. PMID- 20012495 TI - Cognitive impairments in progression of Parkinson's disease. AB - A total of 88 patients with progression of Parkinson's disease (PD) were studied. Cognitive impairments (CI) in PD were in most cases progressive in nature, predominantly because of increases in the severity of dysregulatory and neurodynamic disorders, impairments to visuospatial functions, and, in some cases, deficits in nominative speech function. A high frequency of transformation of moderate cognitive impairments to dementia was demonstrated over periods of 2 5 years. Predictors of the progression of CI in PD were identified: elderly age, later onset of disease, and the severity of PD. The greatest rate of progression of CI was seen in patients with initially more severe impairments of regulatory and visuospatial functions. PMID- 20012496 TI - Immunohistochemical characteristics of the hypophysis in normal conditions and chronic stress. AB - Quantitative immunohistochemical methods were used to assess activation of the hypothalamo-hypophyseal-adrenocortical system at the level of its central component - the adenohypophysis - in the growing body during chronic exposure to psychoemotional stressors of different strengths. Sprague-Dawley rats aged 30 days were subjected to "mild" or "severe" immobilization stress for 5 h per day for seven days. Animals were decapitated at the end of the last stress session and the endocrine glands (hypophysis, adrenals) were harvested, weighed, and embedded in paraffin; sections were stained with hematoxylin and eosin, and also immunohistochemically using monoclonal antibodies to adrenocorticotropic hormone (ACTH) and proliferating cell nuclear antigen (PCNA) following by automated image analysis. These studies showed that stress-associated hyperplasia of corticotropocytes in rats of pubertal age was due more to the differentiation of existing immature precursor cells than to cell proliferation. PMID- 20012497 TI - Stable modifications to the expression of neurohormones in the rat hypothalamus in a model of post-traumatic stress disorder. AB - We report studies of the neuroendocrine mechanisms of development of an anxiety state in rats using the "stress-restress" experimental model of post-traumatic stress disorder. Immunocytochemical methods demonstrated significant increases in corticoliberin expression in both the parvo- and magnocellular parts of the paraventricular nucleus persisting to 10 days after presentation of the animals with repeated stress. Decreases in vasopressin expression were seen in the paraventricular nucleus of the animals on the first day after repeated stress. Vasopressin contents in the parvocellular part of the nucleus in animals of the experimental group were no different at 10 days from those in animals of the control group, while levels in the magnocellular part were increased. These data provide evidence for the involvement of the hypothalamic component of the vasopressinergic system (along with the corticoliberinergic system) in the pathogenetic mechanisms of the analog of post-traumatic stress disorder generated in this model. PMID- 20012498 TI - Immunocytochemical detection of tissue antigens after prolonged storage of specimens in methylsalicylate. AB - The aim of the present work was to verify the possibility of detecting various tissue antigens using standard immunocytochemical methods in paraffin sections of specimens stored in methylsalicylate for two years and longer before embedding in paraffin. Using the immunocytochemical reaction for the intermediate filament proteins nestin, vimentin, and GFAP, the neuron marker NeuN, neuron-specific enolase, and doublecortin as examples, these experiments showed that prolonged storage of dehydrated specimens in methylsalicylate does not prevent detection of antigens using immunocytochemical reactions. PMID- 20012500 TI - Challenges in the study of adjuvant chemoradiation after pancreaticoduodenectomy. PMID- 20012499 TI - Suppression of hypothalamo-hypophyseal-adrenocortical system function as the cause of aggravation of the ulcerogenic action of indomethacin on the stomach after administration of pharmacological doses of hydrocortisone. AB - We report testing of the suggestion that aggravation of the actions of glucocorticoid treatment on the formation of indomethacin-induced erosions may be mediated by inadequate production of glucocorticoid hormones resulting from the suppression of the hypothalamo-hypophyseal-adrenocortical system (HHACS) in these conditions. Administration of indomethacin (25 mg/kg, s.c.) to rats after 24 h of starvation was used to induce gastric erosions. The effects of hydrocortisone given at pharmacological doses on the indomethacin-induced formation of gastric erosions and plasma corticosterone levels were studied one and four weeks after hormone administration. Indomethacin induced increases in plasma corticosterone levels, which were almost completely prevented one week after hydrocortisone administration. The formation of indomethacin-induced erosions was aggravated one week after administration of hydrocortisone, though replacement therapy with corticosterone, which imitates the normal increase in corticosterone, prevented this aggravation. Return of both the increased corticosterone level and the normal sensitivity of the gastric mucosa to the ulcerogenic action of indomethacin occurred four weeks after hydrocortisone administration. These results provide evidence that suppression of HHACS function may be responsible for the aggravation of the action of glucocorticoid treatment on the formation of erosions after administration of indomethacin. PMID- 20012502 TI - Ductoscopy in the evaluation and management of nipple discharge. AB - BACKGROUND: Nipple discharge is a relatively common complaint among patients visiting specialized outpatient clinics for disease of the breast. With advancing technology, it became possible to diagnose and manage nipple discharge using minimally invasive procedures. The aim of this study is to report our experience with ductoscopic evaluation of patients with nipple discharge. MATERIALS AND METHODS: Between September 2005 and February 2009, 236 patients with complaint of nipple discharge were admitted prospectively into the study. All patients were evaluated with ductoscopy. Data concerning age, ductoscopic and postsurgical diagnosis, duration, and complications with the procedure were statistically analyzed. RESULTS: For 236 patients evaluated, there were 249 ductoscopic investigations. Of these, 39 patients were found to have intraductal solitary papilloma of which 24 were excised ductoscopically. Of the 24 ductoscopically excised solitary papillomas, it was possible to remove 17 completely, but 7 papillomas could be removed partially. For 13 patients with solitary papillomas large enough or unfavorably localized so that they could not be snare-excised and for 14 patients with multiple papillomas, we offered surgical removal of the lesions by limited excision of the ducts with the help of ductoscopic localization and marking. There were 35 patients who had debris within the canalicular system, and the debris was washed out. Ductoscopy revealed atypical ductal hyperplasia in 3 patients, ductal carcinoma in situ in 6 patients, and invasive carcinoma in 3 patients; all patients underwent surgery. CONCLUSIONS: We believe that, as the technology quickly improves, ductoscopy will become more widely accepted and applied for breast disorders, not only as a diagnostic tool but also as a privileged therapeutic option for certain pathologies. PMID- 20012501 TI - Gene expression profile prospectively predicts peritoneal relapse after curative surgery of gastric cancer. AB - BACKGROUND: Peritoneal relapse is the most common pattern of tumor progression in advanced gastric cancer. Clinicopathological findings are sometimes inadequate for predicting peritoneal relapse. The aim of this study was to identify patients at high risk of peritoneal relapse in a prospective study based on molecular prediction. METHODS: RNA samples from 141 primary gastric cancer tissues after curative surgery were profiled using oligonucleotide microarrays covering 30,000 human probes. Firstly, we constructed a molecular prediction system and validated its robustness and prognostic validity by 500 times multiple validation by repeated random sampling in a retrospective set of 56 (38 relapse-free and 18 peritoneal-relapse) patients. Secondly, we applied this prediction to 85 patients of the prospective set to assess predictive accuracy and prognostic validity. RESULTS: In the retrospective phase, repeated random validation yielded approximately 68% predictive accuracy and a 22-gene expression profile associated with peritoneal relapse was identified. The prediction system identified patients with poor prognosis. In the prospective phase, the molecular prediction yielded 76.9% overall accuracy. Kaplan-Meier analysis of peritoneal-relapse-free survival showed a significant difference between the "good signature group" and "poor signature group" (log-rank p = 0.0017). Multivariate analysis by Cox regression hazards model identified the molecular prediction as the only independent prognostic factor for peritoneal relapse. CONCLUSIONS: Gene expression profile inherent to primary gastric cancer tissues can be useful in prospective prediction of peritoneal relapse after curative surgery, potentially allowing individualized postoperative management to improve the prognosis of patients with advanced gastric cancer. PMID- 20012503 TI - Posterior ankle impingement: clarification and confirmation of the pathoanatomy. AB - The diagnosis of posterior hindfoot pain may prove to be elusive as there are many potential causes of hindfoot pain, including both soft tissue and osseous abnormalities. This is a case presentation of os trigonum syndrome, with the diagnosis suggested by magnetic resonance imaging (MRI), confirmed by ultrasound, and treated with os trigonum resection. Specific findings of os trigonum syndrome can be seen on MRI, including bone marrow edema pattern and high signal intensity in the synchondrosis as well as in the surrounding soft tissues. Diagnostic injections performed with imaging guidance can help confirm the diagnosis in equivocal cases. Diagnostic and interventional imaging procedures can help confirm the diagnosis of os trigonum syndrome in a patient presenting with hindfoot pain. PMID- 20012504 TI - Challenges for Large Orthopaedic Hospitals Worldwide-An ISOC Position Statement. AB - Since the foundation of the International Society of Orthopaedic Centers in 2006, the group has continued to grow, with currently 11 members worldwide. During the 2008 annual meeting, the bylaws and mission of the group were approved as well as the acceptance of two additional members. Strict inclusion criteria were established to keep the group both small and effective. The goal of the 2008 meeting and this position statement is to identify current challenges in both research and education for large-volume orthopedic hospitals and to discuss possible approaches and solutions. PMID- 20012505 TI - Chaotic neural network applied to two-dimensional motion control. AB - Chaotic dynamics generated in a chaotic neural network model are applied to 2 dimensional (2-D) motion control. The change of position of a moving object in each control time step is determined by a motion function which is calculated from the firing activity of the chaotic neural network. Prototype attractors which correspond to simple motions of the object toward four directions in 2-D space are embedded in the neural network model by designing synaptic connection strengths. Chaotic dynamics introduced by changing system parameters sample intermediate points in the high-dimensional state space between the embedded attractors, resulting in motion in various directions. By means of adaptive switching of the system parameters between a chaotic regime and an attractor regime, the object is able to reach a target in a 2-D maze. In computer experiments, the success rate of this method over many trials not only shows better performance than that of stochastic random pattern generators but also shows that chaotic dynamics can be useful for realizing robust, adaptive and complex control function with simple rules. PMID- 20012506 TI - Long-term follow-up of callotasis lengthening of the capitate after resection of the lunate for the treatment of stage III lunate necrosis. AB - The callotasis lengthening technique was used to gradually lengthen the capitate after resection of the lunate in stage IIIa necrosis in 23 patients. Results of ten patients with a follow-up of at least 5 years showed rapid and sufficient callus formation in every patient regardless of age. The callotasis lengthening modification of the Graner II operation provides all advantages and avoids the major inconvenience of the traditional Graner II operation. There was no increased rate of disturbed fracture healing. Results of the DTPA-gadolinium MRI study did not show any significant impairment of vascularization within the region of the capitate bone. With the "intrinsic bone formation," contrary to every other intercarpal arthrodesis at the wrist, there is no need for an additional bone graft. PMID- 20012507 TI - Reinforcing the staple line during laparoscopic sleeve gastrectomy: prospective randomized clinical study comparing three different techniques. AB - BACKGROUND: Gastric leak and hemorrhage are the most important challenges after laparoscopic sleeve gastrectomy (LSG). In order to reduce these complications, the staple line can be reinforced by absorbable sutures or by the use of glycolide trimethylene carbonate copolymer onto the linear stapler (Gore Seamguard; W.L. Gore & Associates, Inc, Flagstaff, AZ). To our knowledge, there are no randomized studies showing the utility of staple line reinforcement during LSG. The purpose of this study was to randomly compare three techniques in LSG: no staple line reinforcement (group 1), buttressing of the staple line with Gore Seamguard (group 2), and staple line suturing (group 3). METHODS: Between January 2008 and February 2009, 75 patients were prospectively and randomly enrolled in the three different techniques of handling the staple line during LSG. The patient groups were similar (NS). RESULTS: Mean operative time to perform the stomach sectioning was 15.9 +/- 5.9 min (group 1), 20.8 +/- 8.1 min (group 2), and 30.8 +/- 10.1 min (group 3) (p < 0.001). Mean total operative time was 47.4 +/- 10.7 min (group 1), 48.9 +/- 18.4 min (group 2), and 59.9 +/- 19.6 min (group 3) (p = 0.02). Mean blood loss during stomach sectioning was 19.5 +/- 21.3 mL (group 1), 3.6 +/- 4.7 mL (group 2), and 16.7 +/- 23.5 mL (group 3) (p < 0.001). Mean total blood loss was 48.9 +/- 67.1 mL (group 1), 32.5 +/- 46.5 mL (group 2), and 61.9 +/- 69.4 mL (group 3) (p = 0.03). Mean number of stapler cartridges used was 5.6 +/- 0.7 (group 1), 5.7 +/- 0.7 (group 2), and 5.8 +/- 0.6 (group 3) (NS). Postoperative leak affected one patient (group 1), two patients (group 2), and one patient (group 3) (NS). Mean hospital stay was 3.6 +/- 1.4 days (group 1), 3.9 +/- 1.5 days (group 2), and 2.8 +/- 0.8 days (group 3) (p = 0.01). CONCLUSIONS: In LSG, buttressing the staple line with Gore Seamguard statistically reduces blood loss during stomach sectioning as well as overall blood loss. No staple line reinforcement statistically decreases the time to perform stomach sectioning and the total operative time. No significant difference is evidenced in terms of postoperative leak between the three techniques of LSG. PMID- 20012508 TI - Phenotype microarray profiling of Zymomonas mobilis ZM4. AB - In this study, we developed a Phenotype MicroArray (PM) protocol to profile cellular phenotypes in Zymomonas mobilis, which included a standard set of nearly 2,000 assays for carbon, nitrogen, phosphorus and sulfur source utilization, nutrient stimulation, pH and osmotic stresses, and chemical sensitivities with 240 inhibitory chemicals. We observed two positive assays for C-source utilization (fructose and glucose) using the PM screen, which uses redox chemistry and cell respiration as a universal reporter to profile growth phenotypes in a high-throughput 96-well plate-based format. For nitrogen metabolism, the bacterium showed a positive test results for ammonia, aspartate, asparagine, glutamate, glutamine, and peptides. Z. mobilis appeared to use a diverse array of P-sources with two exceptions being pyrophosphate and tripolyphosphate. The assays suggested that Z. mobilis uses both inorganic and organic compounds as S-sources. No stimulation by nutrients was detected; however, there was evidence of partial inhibition by purines and pyrimidines, NAD, and deferoxamine. Z. mobilis was relatively resistant to acid pH, tolerating a pH down to about 4.0. It also tolerated phosphate, sulfate, and nitrate, but was rather sensitive to chloride and nitrite. Z. mobilis showed resistance to a large number of diverse chemicals that inhibit most bacteria. The information from PM analysis provides an overview of Z. mobilis physiology and a foundation for future comparisons of other wild-type and mutant Z. mobilis strains. PMID- 20012509 TI - MDL constrained 3-D grayscale skeletonization algorithm for automated extraction of dendrites and spines from fluorescence confocal images. AB - This paper presents a method for improved automatic delineation of dendrites and spines from three-dimensional (3-D) images of neurons acquired by confocal or multi-photon fluorescence microscopy. The core advance presented here is a direct grayscale skeletonization algorithm that is constrained by a structural complexity penalty using the minimum description length (MDL) principle, and additional neuroanatomy-specific constraints. The 3-D skeleton is extracted directly from the grayscale image data, avoiding errors introduced by image binarization. The MDL method achieves a practical tradeoff between the complexity of the skeleton and its coverage of the fluorescence signal. Additional advances include the use of 3-D spline smoothing of dendrites to improve spine detection, and graph-theoretic algorithms to explore and extract the dendritic structure from the grayscale skeleton using an intensity-weighted minimum spanning tree (IW MST) algorithm. This algorithm was evaluated on 30 datasets organized in 8 groups from multiple laboratories. Spines were detected with false negative rates less than 10% on most datasets (the average is 7.1%), and the average false positive rate was 11.8%. The software is available in open source form. PMID- 20012511 TI - A case of intravascular large B-cell lymphoma of the cutaneous variant: the first case in Asia. PMID- 20012512 TI - Delayed addition of tumor necrosis factor (TNF) antagonists inhibits the generation of CD11c+ dendritic cells derived from CD34+ cells exposed to TNF alpha. AB - We have developed a method that cells exhibiting typical dendritic cell (DC) characteristics are generated from human CD34(+) cells and phagocytose cogenerating erythroid progenitor cells in the presence of tumor necrosis factor alpha (TNF-alpha), interleukin-3, stem cell factor and erythropoietin. Using this system, we titrated the effects of TNF antagonists, etanercept and infliximab, on TNF-alpha activity. We found that 1 microg/ml etanercept dramatically inhibited the generation of CD11c(+) cells accompanying with a complete recovery of the generation of erythroid progenitors. Infliximab at 200 microg/ml exhibited a similar effect to that observed for etanercept. The delayed addition of etanercept to this culture system at day five resulted in significant inhibitory effects on the generation of CD11c(+), CD4(+) and CD86(+) cells. These results indicate that TNF antagonists administered at a concentration that is achievable in vivo, neutralize the biologic effects of TNF-alpha in generating CD11c(+) cells and that a delay in the administration of these antagonists for as long as 5 days partially inhibits the biologic activity of TNF-alpha. These findings may contribute to a great understanding of anti-TNF therapy in patients with an overproduction of cytokines such as hemophagocytic syndromes. PMID- 20012513 TI - Primary cutaneous diffuse large B cell lymphoma relapsed solely as a huge lung tumor mimicking a primary pulmonary lymphoma. AB - Primary cutaneous large B cell lymphoma, leg type (PCLBCL-leg) has recently been identified and recognized as a specific entity. Patients with PCLBCL-leg have a higher relapse rate and a poorer prognosis than the other types of primary cutaneous B cell lymphomas, and disease relapse is confined to the skin in the majority of cases with rare exclusive extracutaneous progression. The late occurrence of lymphoma in patients with a prior history of lymphoma may represent a relapse/progression or a distinct tumor unrelated to the original one. The distinction is of important clinical and therapeutic implications. Here, we report the case of a 90-year-old lady with a history of PCLBCL-leg in complete remission after radiotherapy that developed a huge, solitary pulmonary lymphoma without lymphadenopathy 14 months later. The latter was initially considered as stage IE primary pulmonary lymphoma and was treated with combination chemotherapy resulting in complete remission. Retrospective pathologic review and B cell clonality study revealed that the pulmonary tumor was a diffuse large B cell lymphoma of the same clonal origin as the PCLBCL-leg. This case is unique in the exclusive pulmonary relapse and illustrates the importance of expert pathological review and molecular study in the management of lymphoma patients with unusual clinical features. PMID- 20012515 TI - Diagnostic and clinical benefit of combined coronary calcium and perfusion assessment in patients undergoing PET/CT myocardial perfusion stress imaging. AB - BACKGROUND: A limitation of stress myocardial perfusion imaging (MPI) is the inability to detect non-obstructive coronary artery disease (CAD). One advantage of MPI with a hybrid CT device is the ability to obtain same-setting measurement of the coronary artery calcium score (CACS). METHODS AND RESULTS: Utilizing our single-center nuclear database, we identified 760 consecutive patients with: (1) no CAD history; (2) a normal clinically indicated Rb-82 PET/CT stress perfusion study; and (3) a same-setting CAC scan. 487 of 760 patients (64.1%) had subclinical CAD based on an abnormal CACS. Of those with CAC, the CACS was > or =100, > or =400, and > or =1000 in 47.0%, 22.4%, and 8.4% of patients, respectively. Less than half of the patients with CAC were receiving aspirin or statin medications prior to PET/CT imaging. Patients with CAC were more likely to be initiated or optimized on proven medical therapy for CAD immediately following PET/CT MPI compared to those without CAC. CONCLUSIONS: Subclinical CAD is common in patients without known CAD and normal myocardial perfusion assessed by hybrid PET/CT imaging. Identification of CAC influences subsequent physician prescribing patterns such that those with CAC are more likely to be treated with proven medical therapy for the treatment of CAD. PMID- 20012514 TI - Advances in radionuclide molecular imaging in myocardial biology. AB - Molecular imaging is a new and evolving field that employs a targeted approach to noninvasively assess biologic processes in vivo. By assessing key elements in specific cellular processes prior to irreversible end-organ damage, molecular tools will allow for earlier detection and intervention, improving management and outcomes associated with cardiovascular diseases. The goal of those working to expand this field is not just to provide diagnostic and prognostic information, but rather to guide an individual's pharmacological, cell-based, or genetic therapeutic regimen. This article will review molecular imaging tools in the context of our current understanding of biological processes of the myocardium, including angiogenesis, ventricular remodeling, inflammation, and apoptosis. The focus will be on radiotracer-based molecular imaging modalities with an emphasis on clinical application. Though this field is still in its infancy and may not be fully ready for widespread use, molecular imaging of myocardial biology has begun to show promise of clinical utility in acute and chronic ischemia, acute myocardial infarction, congestive heart failure, as well as in more global inflammatory and immune-mediated responses in the heart-like myocarditis and allogeneic cardiac transplant rejection. With continued research and development, molecular imaging promises to be an important tool for the optimization of cardiovascular care. PMID- 20012517 TI - Digenean parasites of Chinese marine fishes: a list of species, hosts and geographical distribution. AB - In the literature, 630 species of Digenea (Trematoda) have been reported from Chinese marine fishes. These belong to 209 genera and 35 families. The names of these species, along with their hosts, geographical distribution and records, are listed in this paper. PMID- 20012518 TI - Halomonhystera parasitica n. sp. (Nematoda: Monhysteridae), a parasite of Talorchestia brito (Crustacea: Talitridae) in Portugal. AB - Halomonhystera parasitica n. sp. (Monhysterida: Monhysteridae) is described from the body-cavity and under the dorsal plates of the sandy beach amphipod Talorchestia brito Stebbing (Crustacea: Talitridae) in Portugal. The new species differs from previously described members of the genus by a combination of the following characters: four medium-sized cephalic setae; base of stoma with three blunt denticles; posterior dilated portion of stoma absent; amphids small, with width less than quarter of corresponding body width; amphids located less than two labial widths from anterior extremity; uterine eggs elliptical and unembryonated; gubernaculum lacks caudal process; and male tail with two separated pairs of postcloacal papillae and a single subterminal seta-like papilla. This is the first representative of the family Monhysteridae parasitic in the body-cavity of crustaceans. Approximately 48% of the amphipods examined contained various stages of H. parasitica. PMID- 20012519 TI - Ultrastructural features of the tegumental surface of a new metacercaria, Nematostrigea sp. (Trematoda: Strigeidae), with a search for potential taxonomically informative characters. AB - The tegumental surface of a new strigeid metacercaria, Nematostrigea sp., which is a parasite of the freshwater fish Channa gachua (Hamilton) in central Vietnam, is described for the first time using scanning (SEM) and transmission (TEM) electron microscopy. In addition to the general tegumental surface in various parts of the body, details of the surface of the suckers, lappets and holdfast organ are presented, as are variations in the form and distribution of the body spines. As good taxonomic criteria are few in diplostomoid metacercariae at both specific and generic levels, a number of the ultrastructural features revealed may prove to represent taxonomically informative characters. These include the presence of: two rings of dome-shaped papillae localised at different levels on the rim of the oral sucker, a single ring of ciliated papillae on the inner margin of the ventral sucker and a band of dome-shaped papillae along the lateral margins of the broad body-fold in the ventral forebody; an unarmed oral sucker and anteroventral surface of the forebody, although the latter bears protuberant secretory pores; an armed ventral sucker covered by six-pointed spines, except on its rim; multi-pointed spines along the dorsal and ventral sides of the forebody, with the number of their teeth increasing posteriorly; multi-pointed spines on the forebody which gradually transform into single-pointed, more widely distributed spines on the hindbody, disappearing completely at posterior end of the body; the surface of the lappets with a particular distribution of pores leading to three types of secretory glands and three topographical modifications (areas where the surface is smooth, bears digitiform processes or bears recurved, dagger-shaped spines); and the surface of the holdfast organ which is covered with densely packed, straight or slightly curved, simple spines on its lateral surface but is smooth medially. PMID- 20012520 TI - Haemogregarine blood parasites in the lizards Podarcis bocagei (Seoane) and P. carbonelli (Perez-Mellado) (Sauria: Lacertidae) from NW Portugal. AB - In Iberian and Canarian lizards, haemogregarines have been recorded infecting erythrocytes, but most of the records correspond to mature gametocytes. We analysed blood smears from 75 specimens of Podarcis bocagei (Seoane) and 33 specimens of P. carbonelli (Perez-Mellado) from localities of north-western Portugal. We found haemogregarines in 74.7% of P. bocagei and 69.7% of P. carbonelli. Our observations show characteristics of the haemogregarines other than the morphology of the mature gametocytes. In histological sections of the liver of four hosts latent cysts with sporozoites and meronts with merozoites were detected. Both traits have been described as typical of the genera Hepatozoon Miller, 1908 and Hemolivia Petit, Landau, Baccam & Lainson, 1990. We suggest that not only P. bocagei and P. carbonelli from Portugal but other species of Iberian and Canarian lacertids might also be infected by species belonging to one or both genera. PMID- 20012521 TI - Host restriction of HIV-1 by APOBEC3 and viral evasion through Vif. AB - The arms race between virus and host is a constant battle. APOBEC3 proteins are known to be potent innate cellular defenses against both endogenous retroelements and diverse retroviruses. However, retroviruses have developed their own methods to launch counter-strikes. Most primate lentiviruses encode a protein called the viral infectivity factor (Vif). Vif induces targeted destruction of APOBEC3 proteins by hijacking the cellular ubiquitin-proteasome pathway. Here we review the research that led up to the identification of A3G, the mechanisms by which APOBEC3 proteins can inhibit retroelements, and the counter-mechanisms that HIV-1 Vif has developed to evade its antiviral activities. PMID- 20012522 TI - Interactions of viral protein U (Vpu) with cellular factors. AB - Viral protein U (Vpu) is encoded by one of several accessory genes of HIV-1. Like the accessory gene products Vif and Vpr, Vpu targets host proteins such as CD4 for degradation via the recruitment of cellular multi-subunit ubiquitin ligases. Vpu also forms ion channels in cellular membranes. Through one or both of these attributes, Vpu antagonizes host cell factors that restrict the release of progeny virions from infected cells. A key target of Vpu has recently been identified as the interferon-induced transmembrane protein BST-2/CD317 (tetherin), which restricts viral replication by retaining nascent virions on the cell surface. The counteraction of this host defense allows Vpu to be considered an antagonist of the innate immune response to viral infection. PMID- 20012523 TI - TRIM5alpha. AB - TRIM5alpha protein blocks retroviral replication at early postentry stage reducing the accumulation of reverse transcriptase products. TRIM5alpha proteins of Old World primates restrict HIV-1 infection whereas TRIM5alpha proteins of most New World monkeys restrict SIV(mac) infection. TRIM5alpha protein has a RING domain, B-box 2 domain, coiled-coil domain, and PRYSPRY domain. The PRYSPRY domain of TRIM5alpha determines viral specificity and restriction potency by mediating recognition of the retroviral capsid. The coiled-coil domain is essential for TRIM5alpha oligomerization, which contributes to binding avidity for the viral capsid. The RING domain and B-box 2 domain are required for efficient restriction activity of TRIM5alpha protein but the mechanisms remain to be defined. PMID- 20012524 TI - Human immunodeficiency virus type-1 gag and host vesicular trafficking pathways. AB - The Gag protein of HIV-1 directs the particle assembly process. Gag recruits components of the cellular vesicular trafficking machinery in order to traverse the cytoplasm of the cell and reach the particle assembly site. The plasma membrane is the primary site of particle assembly in most cell types, while in macrophages an unusual intracellular membrane-bound compartment bearing markers of late endosomes and the plasma membrane is the predominant assembly site. Plasma membrane specificity of assembly may be directed by components of lipid rafts and the cytoplasmic leaflet component PI(4,5)P(2). Recent work has highlighted the role of adaptor protein complexes, protein sorting and recycling pathways, components of the multivesicular body, and cellular motor proteins in facilitating HIV assembly and budding. This review presents an overview of the relevant vesicular trafficking pathways and describes the individual components implicated in interactions with Gag. PMID- 20012525 TI - The roles of tetraspanins in HIV-1 replication. AB - Tetraspanins are small integral membrane proteins that are known to control a variety of cellular processes, including signaling, migration and cell-cell fusion. Research over the past few years established that they are also regulators of various steps in the HIV-1 replication cycle, but the mechanisms through which these proteins either enhance or repress virus spread remain largely unknown. PMID- 20012526 TI - Imaging of HIV/host protein interactions. AB - HIV-1 relies on a myriad of interactions with host cell proteins to carry out its life cycle. Traditional biochemical approaches to probe protein-protein interactions are limited in their ability to study the spatial and dynamic interactions that take place in the context of an intact cell. However, issues such as localization and dynamics of interactions between viral and host proteins can be well addressed utilizing fluorescent imaging methods. The past decade has brought about the development of many novel fluorescent imaging techniques which have proved useful to describe the interaction of HIV-1 proteins with the host cell. PMID- 20012528 TI - Implications of Nef: host cell interactions in viral persistence and progression to AIDS. AB - The HIV and SIV Nef accessory proteins are potent enhancers of viral persistence and accelerate progression to AIDS in HIV-1-infected patients and non-human primate models. Although relatively small (27-35 kD), Nef can interact with a multitude of cellular factors and induce complex changes in trafficking, signal transduction, and gene expression that together converge to promote viral replication and immune evasion. In particular, Nef recruits several immunologically relevant cellular receptors to the endocytic machinery to reduce the recognition and elimination of virally infected cells by the host immune system, while simultaneously interacting with various kinases to promote T cell activation and viral replication. This review provides an overview on selected Nef interactions with host cell proteins, and discusses their possible relevance for viral spread and pathogenicity. PMID- 20012527 TI - Virological and cellular roles of the transcriptional coactivator LEDGF/p75. AB - The chromatin-associated cellular proteins LEDGF/p75 and LEDGF/p52 have been implicated in transcriptional regulation, cell survival and autoimmunity. LEDGF/p75 also appears to act as a chromatin-docking factor or receptor for HIV-1 and other lentiviruses and to play a role in leukemogenesis. For both the viral and cellular roles of this protein, a key feature is its ability to act as a molecular adaptor and tether proteins to the chromatin fiber. This chapter reviews the emerging roles of LEDGF/p75 and LEDGF/p52 in diverse cellular processes and disease states. PMID- 20012529 TI - Vpr and its interactions with cellular proteins. AB - Like most viral regulatory proteins, HIV-1 Vpr and homologous proteins from primate lentiviruses are small and multifunctional. They are associated with a plethora of effects and functions, including induction of cell cycle arrest in the G(2) phase, induction of apoptosis, transactivation, enhancement of the fidelity of reverse transcription, and nuclear import of viral DNA in macrophages and other nondividing cells. This review focuses on the cellular proteins that have been reported to interact with Vpr and their significance with respect to the known functions and effects of Vpr on cells and on viral replication. PMID- 20012530 TI - Identifying sites of saccade amplitude plasticity in humans: transfer of adaptation between different types of saccade. AB - To view different objects of interest, primates use fast, accurate eye movements called saccades. If saccades become inaccurate, the brain adjusts their amplitudes so they again land on target, a process known as saccade adaptation. The different types of saccades elicited in different behavioral circumstances appear to utilize different parts of the oculomotor circuitry. To gain insight into where adaptation occurs in different saccade pathways, we adapted saccades of one type and examined how that adaptation affected or transferred to saccades of a different type. If adaptation of one type of saccade causes a substantial change in the amplitude of another, that adaptation may occur at a site used in the generation of both types of saccade. Alternatively, if adaptation of one type of saccade transfers only partially, or not at all, to another, adaptation occurs at least in part at a location that is not common to the generation of both types of saccade. We produced significant amplitude reductions in memory-guided, delayed, targeting and express saccades by moving the target backward during the saccade. After memory-guided saccades were adapted, the amplitude of express, targeting and delayed saccades exhibited only a partial reduction. In contrast, when express, targeting, or delayed saccades were adapted, amplitude transfer to memory-guided saccades was more substantial. These results, combined with previously published data, suggest that there are at least two sites of adaptation within the saccadic system. One is used communally in the generation of express, targeting, delayed and memory-guided saccades, whereas the other is specific for the generation of memory-guided saccades. PMID- 20012531 TI - A magnetoencephalography study of choice bias. AB - Many factors can influence, or bias, human decision making. A considerable amount of research has investigated the neural correlates of such biases, mostly correlating hemodynamic responses in brain areas with some aspect of the decision. These studies, typically done using functional magnetic resonance imaging or positron emission tomography, have provided useful information about the location of processing in the brain. However, comparatively little research has examined when these processes occur. The present experiment addressed this question by using magnetoencephalography (MEG) to record brain activity while subjects chose preferred options from decision sets. We found that MEG signal deviations for biased decisions occurred as early as 250-750 ms following stimulus onset. Such deviations occurred earliest in sensors over the right anterior cortex. These findings improve our understanding of temporal dynamics of decision biases and suggest ways that existing explanations for this bias could be refined. PMID- 20012532 TI - Discharge of biceps brachii motor units is modulated by load compliance and forearm posture. AB - The purpose of this study was to compare the discharge characteristics of motor units in the biceps brachii during brief isometric contractions of the elbow flexors as subjects matched either a target force or a target joint angle with the forearm placed in one of two postures. One task required force control to exert a constant force against a rigid restraint (force task), whereas the other task involved position control to maintain a constant elbow angle while supporting an inertial load (position task). The left arm of right-handed subjects was rotated forward so that the upper arm was horizontal and the forearm was vertical. When the elbow flexor muscles were contracted, the wrist exerted a force in a horizontal direction. Subcutaneous electrodes were used to record the discharge of 20 motor units in neutral and supinated forearm postures during both force and position control. Motor unit recruitment thresholds ranged from 1.3 to 37.9% of maximal voluntary contraction force. Discharge rate was similar at the start of the force and position tasks in both the neutral posture (13.1 +/- 0.6 and 12.6 +/- 0.6 pps, P = 0.54) and the supinated posture (14.7 +/- 1.6 and 14.0 +/- 0.9 pps, P = 0.4) and declined during both tasks in the two forearm postures (P < 0.001). Nonetheless, the decrease in discharge rate (P < 0.001), increase in the coefficient of variation for interspike interval (P = 0.04), and increases in the standard deviation of acceleration (P = 0.02) were greatest for the position task in the supinated posture. These findings indicate that the influence of load compliance on the adjustments in motor unit activity during brief isometric contractions with the elbow flexors was modulated by changes in forearm posture. PMID- 20012533 TI - Self versus other in piano performance: detectability of timing perturbations depends on personal playing style. AB - Differences between recorded repetitions of one's own movements are detected more readily than are differences between repetitions of others' movements, suggesting improved visual discrimination due to heightened resonance in the observer's action system and/or relatively accurate internal action simulation (Daprati et al. in Conscious Cogn 16:178-188, 2007). In Experiment 1, we attempted to replicate this finding in the auditory modality. Pianists were recorded playing musical excerpts three times and later judged whether pairs of recordings were the same take or different takes of the same excerpt. They were no better at distinguishing different takes of their own playing than those of other pianists' playing, even though discrimination and self-recognition were well above chance. In Experiment 2, the same pianists tried to detect small local timing deviations that had been introduced artificially. They were better at detecting such deviations in their own performances than in those of another pianist, but only if the deviations were placed at points of a pre-existing self-other difference in local timing. In that case, pianists' ability to predict their own characteristic action pattern did aid their perception of temporal irregularity. These results do not support the perceptual sharpening hypothesis of Daprati et al. in the musical domain, but they do suggest that pianists listening to performances generate idiosyncratic temporal expectations, probably through internal action simulation. PMID- 20012534 TI - Division of labour within the visual system: fact or fiction? Which kind of evidence is appropriate to clarify this debate? AB - The perception versus action hypothesis of Goodale and Milner (Trends Neurosci 15:20-25, 1992) and Milner and Goodale (The visual brain in action. Oxford University Press, Oxford, 1995) postulated two different pathways within the visual system--one for action and one for perception. With the help of pictorial illusions, evidence for this dissociation was found in various studies. There is an ongoing debate, however, as to whether or not this evidence is biased by methodological issues. Indeed, relevant and decisive data can come only from those studies that (1) match conditions appropriately with respect to task demands, (2) use illusions that do not provide any potential obstacles for the hand, (3) do not risk that grasping is either memory driven (when the target is not visible) or online corrected (due to a direct comparison of the grip aperture with the size of the target object), (4) do not confound differences between perception and action conditions with differences in visual feedback, and (5) correct for differences in response functions between grasping and perception. In following all these points outlined above we found support for the perception versus action hypothesis: grip aperture follows actual size independent of illusory effects, while perceived length as indicated by finger-thumb span clearly was subject to the illusion. PMID- 20012535 TI - Roughness of simulated surfaces examined with a haptic tool: effects of spatial period, friction, and resistance amplitude. AB - A specifically designed force-feedback device accurately simulated textures consisting of lateral forces opposing motion, simulating friction. The textures were either periodic trapezoidal forces, or sinusoidal forces spaced at various intervals from 1.5 mm to 8.5 mm. In each of two experiments, 10 subjects interacted with the virtual surfaces using the index finger placed on a mobile plate that produced the forces. The subjects selected their own speed and contact force for exploring the test surface. The apparatus returned force fields as a function of both the finger position and the force normal to the skin allowing full control over the tangential interaction force. In Experiment #1, subjects used an integer, numerical scale of their own choosing to rate the roughness of eight identical, varyingly spaced force ramps superimposed on a background resistance. The results indicated that subjective roughness was significantly, but negatively, correlated (mean r = -0.84) with the spatial period of the resistances for all subjects. In a second experiment, subjects evaluated the roughness of 80 different sinusoidal modulated force fields, which included 4 levels of resistance amplitude, 4 levels of baseline friction, and 5 spatial periods. Multiple regression was used to determine the relationship between friction, tangential force amplitude, and spatial period to roughness. Together, friction and tangential force amplitude produced a combined correlation of 0.70 with subjective roughness. The addition of spatial period only increased the multiple regression correlation to 0.71. The correlation between roughness estimates and the rate of change in tangential force was 0.72 in Experiment #1 and 0.57 in Experiment #2. The results suggest that the sensation of roughness is strongly influenced by friction and tangential force amplitude, whereas the spatial period of simulated texture alone makes a negligible contribution to the sensation of roughness. PMID- 20012536 TI - Fake hands in action: embodiment and control of supernumerary limbs. AB - Demonstrations that the brain can incorporate a fake limb into our bodily representations when stroked in synchrony with our unseen real hand [(the rubber hand illusion (RHI)] are now commonplace. Such demonstrations highlight the dynamic flexibility of the perceptual body image, but evidence for comparable RHI sensitive changes in the body schema used for action is less common. Recent evidence from the RHI supports a distinction between bodily representations for perception (body image) and for action (body schema) (Kammers et al. in Neuropsychologia 44:2430-2436, 2006). The current study challenges and extends these findings by demonstrating that active synchronous stroking of a brush not only elicits perceptual embodiment of a fake limb (body image) but also affects subsequent reaching error (body schema). Participants were presented with two moving fake left hands. When only one was synchronous during active touch, ownership was claimed for the synchronous hand only and the accuracy of reaching was consistent with control of the synchronous hand. When both fake hands were synchronous, ownership was claimed over both, but only one was controlled. Thus, it would appear that fake limbs can be incorporated into the body schema as well as the body image, but while multiple limbs can be incorporated into the body image, the body schema can accommodate only one. PMID- 20012538 TI - The impact of head direction on lateralized choices of target and hand. AB - We examined choices made by monkeys performing a task in which two food-well targets were positioned on either side of the monkey, and LEDs provided instructions on hand use and food target availability. We have previously reported that when gaze and head direction were unrestricted, lateralized choices were biased primarily by hand preference and secondarily by a preference to retrieve a target ipsilateral to the preferred hand. Here, we used a similar behavioral paradigm, but now during trial instructions the monkeys were required to maintain head direction aimed toward a left, a center, or a right fixation LED. When a lateralized head direction was required during presentation of the instructional cues, monkeys were more likely to choose the hand and target ipsilateral to the head direction. Lateralized head direction more strongly biased the monkeys' choice of hand than their choice of target, but hand preference produced even stronger bias on target choices than did head direction. Although target cues were presented before hand cues, our data indicate that target and hand choices were made interactively. We also found that the monkeys' choices were better correlated with their success rate for particular combinations of hand and target than with movement times. PMID- 20012537 TI - Nicotinamide prevents the long-term effects of perinatal asphyxia on apoptosis, non-spatial working memory and anxiety in rats. AB - There is no established treatment for the long-term effects produced by perinatal asphyxia. Thus, we investigated the neuroprotection provided by nicotinamide against the effects elicited by perinatal asphyxia on hippocampus and behaviour observed at 30-90 days of age. Asphyxia was induced by immersing foetuses containing uterine horns, removed from ready-to-deliver rats into a water bath at 37 degrees C for 20 min. Caesarean-delivered siblings were used as controls. Saline or nicotinamide (0.8 mmol/kg, i.p.) was administered to control and asphyxia-exposed animals 24, 48, and 72 h after birth. The animals were examined for morphological changes in hippocampus, focusing on delayed cell death and mossy fibre sprouting, and behaviour, focusing on cognitive behaviour and anxiety. At the age of 30-45 days, asphyxia-exposed rats displayed (1) increased apoptosis, assessed in whole hippocampus by nuclear Hoechst staining, and (2) increased mossy fibre sprouting, restricted to the stratum oriens of dorsal hippocampus, assessed by Timm's staining. Rats from the same cohorts displayed (3) deficits in non-spatial working memory, assessed by a novel object recognition task, and (4) increased anxiety, assessed by an elevated plus-maze test when examined at the age of 90 days. Nicotinamide prevented the effects elicited by perinatal asphyxia on apoptosis, working memory, and anxiety. PMID- 20012540 TI - Two different electron transfer pathways may involve in azoreduction in Shewanella decolorationis S12. AB - Electron transfer pathways for azoreduction by S. decolorationis S12 were studied using a mutant S12-22 which had a transposon insertion in ccmA. The results imply that there are two different pathways for electron transport to azo bonds. The colony of S12-22 was whitish and incapable of producing mature c-type cytochromes whose alpha-peak was at 553 nm in the wild type S12. The mutant S12-22 could not use formate as the sole electron donor for azoreduction either in vivo or in vitro, but intact cells of S12-22 were able to reduce azo dyes of low polarity, such as methyl red, when NADH was served as the sole electron donor. Although the highly polar-sulfonated amaranth could not be reduced by intact cells of S12-22, it could be efficiently reduced by cell extracts of the mutant when NADH was provided as the sole electron donor. These results suggest that the mature c-type cytochromes are essential electron mediators for the extracellular azoreduction of intact cells, while the other pathway without the involvement of mature c-type cytochromes, NADH-dependent oxidoreductase-mediated electron transfer pathway can reduce lowly polar sulfonated azo dyes inside the whole cells or highly polar sulfonated azo dyes in the cell extracts without bacterial membrane barriers. PMID- 20012539 TI - Adaptation of the precision grip orientation to a visual-haptic mismatch. AB - Using a sequential-exposure approach, we examined sensorimotor adaptation of precision grip orientation to a visuo-haptic mismatch. Subjects (n = 41) were asked to grasp with thumb and index finger mirror-viewed objects. They could not see their hand due to the mirror, but they could feel an object located behind the mirror in the same place as the viewed object. When a -15 degrees orientation mismatch between viewed and felt object was introduced, subjects adapted to it by gradually modifying the orientation of their finger opposition axis. When the visuo-haptic mismatch further changed to -30 degrees or to +15 degrees , subjects continued to perform according to their pre-established adaptive state, and gradually modified it until it became adequate for the new mismatch. This outcome supports the existence of a continuously acting mechanism for grasping adaptation, similar to that previously described for pointing and tracking adaptation. Precision grip adaptation generalized to unpracticed object orientations, which suggests that adaptation was based on a sensorimotor recalibration rather than the learning of specific stimulus-response pairs. Transfer to pointing movements was found only when similarity to the grasp adaptation task was high. PMID- 20012541 TI - Agrobacterium tumefaciens-mediated transformation of Withania somnifera (L.) Dunal: an important medicinal plant. AB - This report describes Agrobacterium tumefaciens-mediated transformation of Withania somnifera--an important Indian medicinal plant. A. tumefaciens strain LBA4404, containing the binary vector pIG121Hm was used for transformation, along with the gusA reporter gene with intron under the transcriptional control of the Cauliflower Mosaic Virus (CaMV) 35S promoter. The leaf segments from two-and-a half-month-old green house-grown seedlings were more efficient in transformation, as compared to those from the in vitro-grown shoots. Second expanded leaf from the shoot tip gave the highest transient transformation efficiency. Selection of transgenic shoots was done in the presence of 50 mg l(-1) kanamycin. Polymerase chain reaction analysis of T(0) transgenic plants showed the presence of gusA and nptII genes. The expression of these transgenes in T(1) progeny was confirmed by RT-PCR. The integration of gusA gene was confirmed by Southern blot analysis. The transformation efficiency was found to be 1.67%. PMID- 20012543 TI - Rural environment and risk factors of amyotrophic lateral sclerosis: a case control study. AB - The aetiology of sporadic ALS is still unknown. Links with several environmental factors have been suggested, and some epidemiological studies have shown an increased incidence of ALS in rural populations. This study was designed to investigate risk exposures in a well-delimited rural population and to assess whether rural residency or occupations, such as farming, were associated with an increased risk of developing ALS. A prospective case-control-study of 108 sporadic ALS cases matched by age and sex to 122 controls was performed in Brittany from 2006 to 2008. A strong association was found between agricultural activity and ALS (odds ratio: 2.919; p = 0.01), while rural residence itself did not influence the risk of the disease. Bulbar forms of onset prevailed among agricultural workers as compared with other occupations (55 vs. 26%; p = 0.009). These results suggest a potential role of exposure to agricultural chemicals or contact with animals linked to agricultural work. The prevalence of bulbar forms of onset in agricultural workers has not been reported before. In addition to variable methodological approaches, differences in agricultural practices could explain the discrepancy between these findings and other studies. PMID- 20012544 TI - The thalamic ultrastructural abnormalities in paroxysmal kinesigenic choreoathetosis: a diffusion tensor imaging study. AB - Paroxysmal kinesigenic choreoathetosis (PKC) is a rare neurologic disorder. There are not apparent morphological changes in patients with idiopathic PKC. The purpose of this study is to determine whether ultrastructural changes are in the brain of patients with idiopathic PKC using diffusion tensor imaging. From May 2007 to August 2008, seven patients with idiopathic PKC were included. The mean age at initial onset was 11.7 +/- 3.1 (range 8-17) years, and the mean disease duration was 6.9 +/- 5.1 (range 1-14) years. Seven subjects of an age- and sex matched control group were recruited. DTI data were obtained with a 3-T scanner. Fractional anisotropy (FA) and mean diffusivity (MD) were obtained in eight brain regions of interest. Patients with idiopathic PKC had significantly higher FA values than controls in the right thalamus (P < 0.05 Bonferroni corrected). Patients also had lower MD values than controls in the left thalamus (P < 0.05 Bonferroni corrected). FA and MD values were not significantly correlated with age of onset, gender, frequency of attack and duration of the disease. The results showed that in patients with idiopathic PKC, diffusion tensor imaging discloses distinct ultrastructural abnormalities in the thalamus. DTI is a sensitive neuroradiologic technique for detecting cerebral alterations in patients even without visible lesions on conventional MRI. PMID- 20012542 TI - Molecular genetics of congenital atrial septal defects. AB - Congenital heart defects (CHD) are the most common developmental errors in humans, affecting 8 out of 1,000 newborns. Clinical diagnosis and treatment of CHD has dramatically improved in the last decades. Hence, the majority of CHD patients are now reaching reproductive age. While the risk of familial recurrence has been evaluated in various population studies, little is known about the genetic pathogenesis of CHD. In recent years significant progress has been made in uncovering genetic processes during cardiac development. Data from human genetic studies in CHD patients indicate that the genetic aetiology was presumably underestimated in the past. Inherited mutations in genes encoding cardiac transcription factors and sarcomeric proteins were found as an underlying cause for familial recurrence of non-syndromic CHD in humans, in particular cardiac septal defects. Notably, the cardiac phenotypes most frequently seen in mutation carriers are ostium secundum atrial septal defects (ASDII). This review outlines experimental approaches employed for the detection of CHD-related genes in humans and summarizes recent findings in molecular genetics of congenital cardiac septal defects with an emphasis on ASDII. PMID- 20012545 TI - Bistable dynamics of cardiac cell models coupled by dynamic gap junctions linked to cardiac memory. AB - In an earlier study, we suggested that adaptive gap junctions (GJs) might be a basis of cardiac memory, a phenomenon which refers to persistent electrophysiological response of the heart to external pacing. Later, it was also shown that the proposed mechanism of adaptation of GJs is consistent with known electrophysiology of GJs. In the present article, we show that a pair of cardiac cell models coupled by dynamic, voltage-sensitive GJs exhibits bistable behavior under certain conditions. Three kinds of cell pairs are considered: (1) a Noble Noble cell pair that represents adjacent cells in Purkinje network, (2) a pair of DiFranceso-Noble cells that represents adjacent SA nodal cells, and (3) a model of Noble cell coupled to Luo-Rudy cell model, which represents an interacting pair of a Purkinje fiber and a ventricular myocyte. Bistability is demonstrated in all the three cases. We suggest that this bistability might be an underlying factor behind cardiac memory. Focused analysis of a pair of Noble cell models showed that bistability is obtained only when the properties of GJs "match" with the properties of the pair of cells that is coupled by the GJs. This novel notion of match between GJs and cardiac cell types might give an insight into specialized distributions of various connexin proteins in cardiac tissue. PMID- 20012546 TI - Density-dependence of functional development in spiking cortical networks grown in vitro. AB - During development, the mammalian brain differentiates into specialized regions with distinct functional abilities. While many factors contribute to functional specialization, we explore the effect of neuronal density on the development of neuronal interactions in vitro. Two types of cortical networks, namely, dense and sparse with 50,000 and 12,500 total cells, respectively, are studied. Activation graphs that represent pairwise neuronal interactions are constructed using a competitive first response model. These graphs reveal that, during development in vitro, dense networks form activation connections earlier than sparse networks. Link entropy analysis of dense network activation graphs suggests that the majority of connections between electrodes are reciprocal in nature. Information theoretic measures reveal that early functional information interactions (among three electrodes) are synergetic in both dense and sparse networks. However, during later stages of development, previously synergetic relationships become primarily redundant in dense, but not in sparse networks. Large link entropy values in the activation graph are related to the domination of redundant ensembles in late stages of development in dense networks. Results demonstrate differences between dense and sparse networks in terms of informational groups, pairwise relationships, and activation graphs. These differences suggest that variations in cell density may result in different functional specializations of nervous system tissue in vivo. PMID- 20012547 TI - Radiosensitive gene therapy through imRNA expression for silencing manganese superoxide dismutase. AB - BACKGROUND: In radiotherapy, therapeutic efficacy for the control of cancer is often limited by tumor tissue radioresistance including nasopharyngeal carcinoma. Exposure of cells to ionizing radiation (IR) leads to the formation of reactive oxygen species that are associated with radiation-induced cellular apoptosis and necrosis. The antioxidant enzyme manganese superoxide dismutase (SOD2) catalyzes the dismutation of the superoxide anions into hydrogen peroxide. METHODS: We have investigated the potential of SOD2 gene silencing, through plasmid transfer using a microRNA interference optimized for transcription in nasopharyngeal carcinoma cell line cells, to degrade the radioresistance of a human nasopharyngeal carcinoma cell line. Using these as in vitro models we have investigated whether SOD2 gene therapy may be suitable for the reduction of the nasopharyngeal carcinoma resistance to the effects of IR. RESULTS: Here we demonstrate using both biological and physical assays that silencing of SOD2 enhances the radiosensitivity of nasopharyngeal carcinoma cells to IR injury. Our results show that a decrease in the levels of SOD2 mRNA and protein within CNE1 cells (down regulated 65 and 80%) leads to a significant decrease in clonogenic survival (from 24.5 to 9.67% at 2 Gy, from 9.12 to 2.45% at 4 Gy), as evident by a significant decrease in Dbar (from 1.923 to 0.617 Gy), SF(2) (from 0.403 to 0.021) values, and a significant increase in the alpha value (from 0.228 +/- 0.070 to 1.064 +/- 0.210/Gy) when compared either to cells transduced with a Gateway-adapted expression vector encoding EmGFP alone or to the parental line. CONCLUSIONS: The results presented suggest that miRNA for silencing SOD2 radiosensitizing gene therapy maybe applicable to the nasopharyngeal carcinoma, improving the therapeutic ratio of cancer radiotherapy. PMID- 20012548 TI - EBP50 exerts tumor suppressor activity by promoting cell apoptosis and retarding extracellular signal-regulated kinase activity. AB - The expression of Ezrin-radixin-moesin-binding phosphoprotein-50 (EBP50) and the intragenic mutation of the ebp50 gene have been reported to correlate with human breast cancer development, but the exact impacts on breast cancer development and its molecular mechanism are not fully understood. In this study, we investigate the potential function of EBP50 through over-expression in the breast cancer cell line, MDA-MB-231, which has low EBP50 protein expression levels. The effects of EBP50 over-expression on cellular proliferation, anchorage-independent growth and apoptosis were examined. In addition, the activity of extracellular signal regulated kinase (ERK) was also determined. Our results show that a decrease of cellular proliferation and attenuation of colony-forming ability were evident in MDA-MB-231 cells stably transfected with an EBP50 expressing plasmid (EBP-231) when compared with control cells. There was also a statistically significant increase in spontaneous apoptosis in EBP-231 cells accompanied by an attenuation in ERK activity. Altogether, our results suggest that restoring EBP50 expression could suppress breast cancer cell proliferation by promoting cell apoptosis and inhibiting ERK activity, and that EBP50 may be a target for development of diagnostics and therapeutics in breast cancer. PMID- 20012549 TI - Anti-proliferative and cytotoxic activity of pentadactylin isolated from Leptodactylus labyrinthicus on melanoma cells. AB - Nowadays, the emergence of resistance to the current available chemotherapeutic drugs by cancer cells makes the development of new agents imperative. The skin secretion of amphibians is a natural rich source of antimicrobial peptides (AMP), and researchers have shown that some of these wide spectrum molecules are also toxic to cancer cells. The aim of this study was to verify a putative anticancer activity of the AMP pentadactylin isolated for the first time from the skin secretion of the frog Leptodactylus labyrinthicus and also to study its cytotoxic mechanism to the murine melanoma cell line B16F10. The results have shown that pentadactylin reduces the cell viability of B16F10 cells in a dose-dependent manner. It was also cytotoxic to normal human fibroblast cells; nevertheless, pentadactylin was more potent in the first case. The studies of action mechanism revealed that pentadactylin causes cell morphology alterations (e.g., round shape and shrinkage morphology), membrane disruption, DNA fragmentation, cell cycle arrest at the S phase, and alteration of mitochondrial membrane potential, suggesting that B16F10 cells die by apoptosis. The exact mechanism that causes reduction of cell viability and cytotoxicity after treatment with pentadactylin is still unknown. In conclusion, as cancer cells become resilient to death, it is worthwhile the discovery of new drugs such as pentadactylin that induces apoptosis. PMID- 20012550 TI - The effect of anti-TNF agent on oxidation status in patients with ankylosing spondylitis. AB - The aim of this study was to comparatively evaluate oxidative status of ankylosing spondylitis (AS) patients receiving anti-tumor necrosis factor (TNF) or non-steroid anti-inflammatory drugs (NSAID). Forty-seven patients with AS and 27 healthy controls were enrolled. Of these, 23 were on anti-TNF (group 1) and 24 on NSAIDs (group 2). Groups 1 and 2 were consisted of matched patients with respect to age, gender, body mass index, disease duration, C-reactive protein, erythrocyte sedimentation rate, total cholesterol, and Bath Ankylosing Spondylitis Disease Activity Index. Mean duration of treatment for patients in group 1 was 12.6 +/- 6.8 months. Serum total antioxidative status (TAS) and total oxidative status (TOS) levels were determined using new automated methods. Oxidative stress index (OSI) was calculated. The groups' carotid intima-media thicknesses (IMT-C) were also measured using ultrasonography. Group 1 had the highest TAS and lowest TOS levels. The TOS levels of group 1 was lower than the control, while group 2 being higher than controls. The difference in TOS levels between group 1 and group 2 was statistically significant (p = 0.040). OSI values were highest in group 2 and lowest in group 1. There was no significant correlation between oxidant/antioxidant parameters and IMT-C for group 1 (r = 0.30, p = 0.198 for OSI; r = 0.22, p = 0.366 for TAS; r = -0.22, p = 0.361 for TOS). This is the first study to evaluate total oxidative/antioxidative status in patients with AS on anti-TNF agent. These results clearly indicate positive effects of anti-TNF treatment on oxidative status of AS patients. The limited effects of NSAIDs compared with controls may be due to excess impaired oxidative status in the patients in this study. PMID- 20012551 TI - Headache associated with moyamoya disease: a case story and literature review. AB - Headache associated with moyamoya disease (HAMD) is common in moyamoya disease. However, the characteristics and classification of HAMD are largely unknown. We present a case of a 39-year-old woman with HAMD. To characterize and classify the features of this syndrome, the patient was asked to complete a 4-month diagnostic headache diary. There was a total of 15 ictal days. All episodes were without aura. The headache was more commonly pressing (10/15), mild to moderate in severity (14/15), unchanged by physical activity (11/15), and associated with photophobia (10/15). The International Headache Society Classification was utilized to determine that eight episodes met criteria for probable migraine without aura, while seven episodes met criteria for probable frequent episodic tension-type headache. We identified four other case reports of HAMD with partial descriptions of the characteristics. When combined with our patient, the median age was 34 years old (range 6-49, SD 16). Four were female, while the patient with cluster headache was male. The median time from headache onset to diagnosis with moyamoya disease was 9.5 months (range 0-192, SD 88.0). Headaches were described as migraine with aura in two of five cases, hemiplegic migraine in one of five, and cluster headache in one of five. The highest intensity was described as severe in three of three cases, in which headache intensity was reported. Meanwhile, nausea, vomiting, and photophobia were present in two of three cases, where these features were reported, while nausea without vomiting was seen in one of three cases. In all five cases, patients had other neurological symptoms, such as paresis, seizures, visual disturbances, dysarthria, allodynia, ptosis, and unilateral restless leg syndrome. In conclusion, HAMD can present as migraine without aura. It can be the first presenting symptom of moyamoya disease. The headache features are not diagnostic; hence, early neurovascular imaging should be considered in patients with new onset, refractory migraine-like headache, especially in the setting of other neurological symptoms to exclude underlying moyamoya disease. Further reports using headache diaries are needed to better characterize HAMD as well as to determine whether headache with tension-type features is also part of this condition. PMID- 20012552 TI - Analysing overexpression of L-valine biosynthesis genes in pyruvate-dehydrogenase deficient Corynebacterium glutamicum. AB - L-valine biosynthesis was analysed by comparing different plasmids in pyruvate dehydrogenase-deficient Corynebacterium glutamicum strains in order to achieve an optimal production strain. The plasmids contained different combinations of the genes ilvBNCDE encoding for the L-valine forming pathway. It was shown that overexpression of the ilvBN genes encoding acetolactate synthase is obligatory for efficient pyruvate conversion and to prevent L-alanine as a by-product. In contrast to earlier studies, overexpression of ilvE encoding transaminase B is favourable in pyruvate-dehydrogenase-negative strains. Its amplification enhanced L-valine formation and avoided extra- and intracellular accumulation of ketoisovalerate. PMID- 20012553 TI - Copper(II) complexes of methimazole, an anti Grave's disease drug. Synthesis, characterization and its potential biological behavior as alkaline phosphatase inhibitor. AB - Methimazole (MeimzH) is an anti-thyroid drug and the first choice for patients with Grave's disease. Two new copper(II) complexes of this drug: [Cu(MeimzH)(2)(NO(3))(2)]*0.5H(2)O and [Cu(MeimzH)(2)(H(2)O)(2)](NO(3))(2)*H(2)O were synthesized and characterized by elemental analysis, dissolution behavior, thermogravimetric analysis and UV-vis, diffuse reflectance, FTIR and EPR spectroscopies. As it is known that copper(II) cation can act as an inhibitor of alkaline phosphatase (ALP), the inhibitory effect of methimazole and its copper(II) complexes on ALP activity has also been investigated. PMID- 20012554 TI - Association of CTLA4 gene polymorphism in Iranian patients with ankylosing spondylitis. AB - INTRODUCTION: Cytotoxic T-lymphocyte antigen 4 (CTLA-4) is a costimulatory molecule expressed by activated T cells. This study was performed to investigate the allele and genotype frequencies of CTLA4 gene polymorphisms in Iranian patients with ankylosing spondylitis (AS). METHODS: One hundred and fifty-seven patients with AS and 103 controls were included in this study. Polymorphisms of CTLA4 gene at positions +49 (in exon 1), -318, and -1,147 (in the promoter region) were studied on the genomic DNA using PCR restriction fragment-length polymorphism method. RESULTS: The frequencies of the T allele at position -1147 in the patients with AS was significantly increased in comparison with the control group (11% vs. 5%, P = 0.004); whereas the frequencies of C allele at the same position were significantly decreased in the patient group (89% vs. 95%, P = 0.004). Comparison of genotype frequencies at this position showed that the frequency of CT genotype in comparison with other genotypes was overrepresented in the patient group (20% vs. 8%, P = 0.012), while the CC genotype in comparison with other genotypes was decreased (79% vs. 91%, P = 0.012). There was no significant difference on frequencies of genotypes at the positions -318 and +49. CONCLUSION: This study could suggest an association between specific allele in the promoter region of CTLA4 gene and AS disease. PMID- 20012555 TI - Effects of alcohol on trajectories of physical aggression among urban youth: an application of latent trajectory modeling. AB - Several studies have investigated factors associated with physical aggression during adolescence. Yet, little is known about the longitudinal relationship between drug use, particularly alcohol use, and physical aggression among minority youth. The present study examined the effects of alcohol and substance use at age 11 on trajectories of physical aggression over time (ages 12-14) among urban adolescents from Chicago, IL. Data from the Project Northland Chicago (n = 3038, 49.4% female) was used. The current study sample included 1,160 Black, 1,015 Hispanic and 863 White/other adolescents for a total of 3,038 adolescents. Four trajectories of physical aggression were identified: Non-aggressive (16%), Desistors (9%), Escalators (20%) and Chronic Aggressive (55%). After adjusting for physical aggression behaviors, delinquent friends, lack of supervised time, demographic variables, smoking and marijuana use, past year alcohol users at age 11 were 2.1 times more likely to be "Escalators" and 1.9 times more likely to be in the "Chronic Aggressive" group. Gender and ethnic differences were also observed in the trajectories of physical aggression. Black youth were 2.5 times more likely to be in the "Chronic Aggressive" group. Findings highlight the importance of targeting alcohol prevention to reduce physical aggression among urban young adolescents. PMID- 20012556 TI - Using QEEG-guided neurofeedback for epilepsy versus standardized protocols: enhanced effectiveness? AB - This article briefly reviews some of the past EEG treatments of epilepsy and discusses how QEEG may potentially enhance effectiveness of this approach. Several cases are presented in support of this approach. PMID- 20012557 TI - Editor's note: focus on QEEG...and EEG. PMID- 20012558 TI - Achievement of targeted posterior slope in the medial opening wedge high tibial osteotomy: a mathematical approach. AB - High tibial osteotomy (HTO) for medial compartment knee osteoarthritis is preferred in the activity patient since it allows patients to return to sports and recreational activities similar to the preoperative level. The purpose of this study was to mathematically formulate medial and anteromedial opening gaps in the medial opening wedge HTO to achieve a targeted tibial posterior slope. The change of posterior slope angle was mathematically derived in terms of the medial and anteromedial opening gaps, and the medial opening angles. The derived equations were validated by comparing them with those directly measured by performing simulated HTOs. In the triangular geometries of osteotomy planes, measured from three-dimensional osteotomy models of 30 knee patients, the mean anteromedial, medial, and lateral included angles were 92.4 degrees, 53.9 degrees, and 33.7 degrees, respectively, and the mean lateral-medial edge length was 53.3 mm. The ratio of the anteromedial opening gap to the posterior opening gap should be "sin(the medial included angle)xcos(the lateral included angle)/sin(the anteromedial included angle)" to maintain an intact posterior tibial slope angle. With the derived equations, surgeons can estimate the opening gaps and opening angles to get a targeted posterior tibial slope with a medial opening angle. PMID- 20012559 TI - Implications of available design space for identification of non-immunogenic protein therapeutics. AB - Immunogenicity/antibody responses are major issues for parenteral proteins and nanotherapeutics (nanovectors, diagnostics, theranostics, etc.), and robust antibody responses require T-helper epitopes. T-helper epitopes consist of specific amino acids at specific positions (anchor positions) in immunogens which contact the major histocompatibility complex (MHC), provide most of the energy for MHC binding and constitute the binding motif for the corresponding MHC alleles. We developed an algorithm that considers motifs to design vaccines lacking unwanted T-cell epitopes, and found numbers of such vaccines can be astronomical (Lee et al. 2009). The algorithm can be used to design reduced immunogenicity proteins, and numbers of predicted proteins are also immense. Reducing T-helper epitope content reduces protein immunogenicity, but the depth of mutagensis needed to eliminate immunogenicity is commonly assumed to be too great for retention of protein bioactivity. However, very deep, but successful substitution, insertion and deletion mutagenesis have been reported. These reports and design space the algorithm reveals suggest development of non immunogenic therapeutics might be more feasible than commonly assumed. PMID- 20012560 TI - Measurement of retinal vascular tortuosity and its application to retinal pathologies. AB - The tortuosity of retinal blood vessels is an important diagnostic indicator for a number of retinal pathologies. We applied robust quantitative tortuosity metrics, which are well suited to automated detection and measurement, to retinal fluorescein images of normal and diseased vessels exhibiting background diabetic retinopathy, retinitis pigmentosa and retinal vasculitis. We established the validity of the mean tortuosity (M) and the normalized root-mean-square tortuosity (K) by their strong correlation with the ranking of tortuosity by an expert panel of ophthalmologists. The low prevalences of the diseased conditions in the general population affect the classification process, and preclude the use of tortuosity for screening for all of these conditions simultaneously in the general population. Tortuosity may be useful as a screening test for retinitis alone, and may be useful for distinguishing diabetic retinopathy or vasculitis from normal in a discretionary (i.e. referred) population. PMID- 20012561 TI - Racial disparities in the quality of medication use in older adults: baseline findings from a longitudinal study. AB - BACKGROUND: Medication-related problems are prevalent in older adults and adversely affect the quality of care. It has been suggested that racial differences exist in medication use. Most efforts to evaluate the quality of medication use target specific drugs or disease states, or a set of pre-defined quality indicators, rather than the patient. OBJECTIVE: We conducted a prospective cohort study to determine the prevalence and types of medication related problems in older adults, examining the impact of race on quality medication use. METHODS: In-home interviews and medical record reviews of 200 (100 white, 100 black) older adults were conducted three times over 1 year. The quality of medication use was measured using a clinical pharmacist's assessment of quality and the Assessing Care of Vulnerable Elders quality indicators. We used logistic and negative binomial regression models to analyze the two primary endpoints of prevalence and number of medication-related problems. RESULTS: Mean age was 78.3 (whites) and 75.5 (blacks), with the majority being female. Although whites used more medications than blacks (11.6 versus 9.7; p < 0.01), blacks had more medication-related problems per person than whites (6.2 versus 4.9; p < 0.01). All patients had at least one medication-related problem; undertreatment, suboptimal drug, suboptimal dosing, and nonadherence were most prevalent. Blacks had significantly higher rates of nonadherence than whites (68% versus 42%; p < 0.01). CONCLUSION: Medication-related problems are prevalent in community residing older adults. Blacks had more medication-related problems than whites, including higher rates of nonadherence. These findings require further study to better understand racial disparities in quality medication use. PMID- 20012562 TI - Notch1 expression correlates with tumor differentiation status in ovarian carcinoma. AB - Notch signaling and its receptor Notch1 are expressed in ovarian epithelial tumors, but the relationship between Notch signaling and ovarian cancer remains to be elucidated. In this study, we detected the expression of Notch1 in ovarian tissues and human ovarian cancer cell lines. We also analyzed the expression of Notch1 and its relationship with differentiation status and FIGO (Federation International of Gynecology and Obstetrics) stage in ovarian cancer tissues. Immunohistochemistry, real-time polymerase chain reaction and Western blot were used to detect the expression of Notch1 in 109 ovarian cancer tissues, 65 patient matched opposite side normal ovarian tissues and 48 normal ovarian tissues, together with A2780, HO-8910 and IOSE 144 cell lines. Our results showed that the expression of Notch1 in ovarian cancer tissues was higher than that in matched normal tissues and normal tissues, Notch1 is highly expressed in ovarian cancer cells A2780 and HO-8910. Moreover, expression of Notch1 increased gradually with the poor differentiating of cancer tissues and the increasing of FIGO stage in ovarian cancer tissues. It was concluded that Notch1 might be involved and play an oncogenic role in the development of ovarian cancer. PMID- 20012563 TI - Future directions in immunomodulatory therapy. AB - The role of immunomodulatory-based therapy with thalidomide or lenalidomide is clearly established in the management of patients with myeloma in all phases of their disease. Recent preclinical and clinical works have demonstrated that in addition to combination therapy with dexamethasone, there is significant activity when combined with the proteasome inhibitor bortezomib. More recent clinical studies have also demonstrated significant activity when combined with akt inhibitors, HDAC inhibitors, and even monoclonal antibodies. Further clinical development of immunomodulatory agents should continue to be based on preclinical rationale, which has resulted in a number of promising and clinically active combinations. PMID- 20012565 TI - [Health risks: assessment, knowledge, and perceptions ]. PMID- 20012564 TI - Diagnostic value of fibronectin and mutant p53 in the urine of patients with bladder cancer: impact on clinicopathological features and disease recurrence. AB - Development of new methods for bladder cancer detection is required because cystoscopy is invasive, and voided urine cytology (VUC) has low sensitivity. The aim of this study was to evaluate the diagnostic performance of urinary fibronectin and mutant p53 in comparison with VUC in the detection of bladder cancer. This study included 100 patients diagnosed with bladder cancer, 93 patients with benign urological disorders and 47 healthy volunteers. The urine supernatant was used for determination of fibronectin by ELISA, while urine sediment was used for cytology and detection of mutant p53 by PCR-SSCP followed by DNA sequencing. The sensitivity and specificity were 59% and 91.4% for VUC, 82% and 84.3% for fibronectin, and 37% and 100% for mutant p53; combination of the three parameters increased sensitivity to 95% but specificity was only 78.6%. A significant association was observed between disease recurrence and mutant p53, stage and lymph node involvement. Our results indicate that fibronectin had the highest sensitivity compared to VUC and mutant p53 in bladder cancer detection; however, mutant p53 had superior specificity compared to VUC and fibronectin. Mutant p53 is associated with disease recurrence and hence it has a significant prognostic role in bladder cancer. PMID- 20012566 TI - [Epidemiological and statistical methods of risk assessment]. AB - Risk assessment offers a variety of methods which comprise different risk concepts to assess population-based or individual risks. A central epidemiological measure of risk is incidence which is used to derive the relative risk as the ratio between the incidence among subjects exposed and the incidence among subjects unexposed to a certain risk factor. The excess risk quantifies the risk of an exposed population that exceeds the background risk of a non-exposed population. In environmental epidemiology the latter is frequently based on a constant exposure dose over a certain period and is expressed as unit risk. Since the application of different risk concepts depends on the respective study type, we first discuss the different epidemiological study designs. Next, methods to derive the different risk concepts are evaluated and the corresponding statistical measures are presented. Finally, an example illustrates the application of the discussed concepts for individual risk assessment. PMID- 20012570 TI - [Maintenance of the supply of blood products in the event of an influenza pandemic]. PMID- 20012567 TI - [Multimorbidity and health-related quality of life among elderly persons]. AB - Aim of this study was to analyze the effects of single diseases and multimorbidity on health-related quality of life (HRQoL) in the elderly. Based on data from telephone interviews with the getABI cohort, empirical analyses were conducted. To evaluate HRQoL, 2,120 participants (76.29 +/- 4.48 years old, 46.3% male) completed the EQ-5D and the SF-8 questionnaire. There is an inverse relationship between HRQoL and multimorbidity in the elderly. In comparison to psychological dimensions, physical domains decrease more clearly with rising morbidity. Analyzing the influence of single diseases, particularly cardiovascular and cerebrovascular diseases diagnosed in the last two years, the long-term existence of peripheral arterial disease and dysfunctions of the musculoskeletal system result in severe loss of HRQoL in older persons. Information on HRQoL can help health care providers obtain a comprehensive picture of their older and often multimorbid patients' state and perception of health. An optimized health care process should not only focus on the individual diagnoses, but also on the extent of multimorbidity and associated HRQoL effects. PMID- 20012571 TI - [12th notification of changes to the list of material preparation and disinfection processes pursuant to--11th drinking water regulation 2001 Standard: December 2009]. PMID- 20012572 TI - [Addendum on the "Environmental monograph: Mercury--reference-- and human biomonitoring values" of the Human Biomonitoring Commission of the Environmental office. Position of the Human Biomonitoring commission of the Environmental Office]. PMID- 20012574 TI - Coming full circle: cyclin-dependent kinases as anti-cancer drug targets. AB - Because the normal control of cell proliferation is disturbed in cancer, the cyclin-dependent kinases (CDKs) that trigger DNA synthesis and mitosis have been popular targets for inhibition with small molecules, but the jury is still out on whether this will be an effective anti-tumor strategy. There is debate about which of the multiple CDKs active during the mammalian cell cycle might be good targets, reflecting fundamental confusion about what, precisely, those different CDKs really do. In the classical view, based largely on their activation timing in cycling cells, different CDKs are specialized to perform discrete functions during distinct cell-cycle intervals. A revisionist model has emerged in which all functions essential to cell division can be performed by a single catalytic subunit, based on the ability of cells to proliferate and animals to survive when individual CDKs are removed by gene deletion or depleted by RNA interference. That those situations in no way resemble ones in which CDKs are inhibited pharmacologically is often overlooked or downplayed. A more nuanced - and accurate - picture is now coming into view, thanks to recent studies that reveal kinetically distinct pathways of activation for closely related CDKs and CDK specific roles in the temporal control of S phase. The basic question of whether CDKs can be effectively targeted in cancer has yet to be answered but can now be addressed in chemical-genetic model systems that approximate the situation - still hypothetical - of truly selective CDK inhibition in vivo. PMID- 20012575 TI - Core and linker histone modifications involved in the DNA damage response. AB - The stability of the genome is constantly under attack from both endogenous and exogenous DNA damaging agents. These agents, as well as naturally occurring processes such as DNA replication and recombination can result in DNA double strand breaks (DSBs). DSBs are potentially lethal and so eukaryotic cells have evolved an elaborate pathway, the DNA damage response, which detects the damage, recruits proteins to the DSBs, activates checkpoints to stall cell cycle progression and ultimately mediates repair of the damaged DNA. As the DSBs occur in the context of chromatin, execution of this response is partly orchestrated through the modification of the DNA-bound histone proteins. These histone modifications include the addition or removal of various chemical groups or small peptides and function to change the chromatin structure or to attract factors involved in the DNA damage response, and as such, are particularly important in the early stages of the DNA damage response. This review will focus on such modifications, the enzymes responsible and also highlights their importance by reporting known roles for these modifications in genome stability and disease. PMID- 20012576 TI - Chromatin assembly and signalling the end of DNA repair requires acetylation of histone H3 on lysine 56. AB - The packaging of DNA into chromatin results in a barrier to all DNA transactions. To facilitate transcription, replication and repair histone proteins are frequently post-translational modified. Such covalent additions to histone residues can modulate chromatin folding and/or provide specificity to docking surfaces for non-histone chromatin proteins. In the budding yeast, one such modification, transient acetylation of histone H3 on residue lysine 56 (H3K56ac); occurs on newly synthesized H3 molecules and facilitates their deposition onto newly replicated DNA during S phase. H3K56ac also has a role in chromatin reassembly following DNA damage in S phase. Importantly, the completion of H3K56ac-dependent chromatin reassembly appears to be required for resumption of cell proliferation after DNA repair. Emerging evidence, although not without conflict, suggests that H3K56ac is not only present in human cells, but is similarly regulated and required for chromatin reassembly. PMID- 20012577 TI - Structure and function of histone H2AX. AB - Histone H2AX is a histone variant found in almost all eukaryotes. It makes a central contribution to genome stability through its role in the signaling of DNA damage events and by acting as a foundation for the assembly of repair foci. The H2AX protein sequence is highly similar and in some cases overlapping with replication-dependent canonical H2A, yet the H2AX gene and protein structures exhibit a number of features specific to the role of this histone in DNA repair. The most well known of these is a specific serine at the extreme C-terminus of H2AX which is phosphorylated by Phosphoinositide-3-Kinase-related protein Kinases (PIKKs) to generate the gammaH2AX mark. However, recent studies have demonstrated that phosphorylation, ubiquitylation and other post-translational modifications are also crucial for function. H2AX transcript properties suggest a capability to respond to damage events. Furthermore, the biochemical properties of H2AX protein within the nucleosome structure and its distribution within chromatin also point to features linked to its role in the DNA damage response. In particular, the theoretical inter-nucleosomal spacing of H2AX and the potential implications of amino acid residues distinguishing H2AX from canonical H2A in structure and dynamics are considered in detail. This review summarises current understanding of H2AX from a structure-function perspective. PMID- 20012578 TI - The initiation step of eukaryotic DNA replication. AB - Eukaryotic initiation of DNA replication is a tightly regulated process. In the yeasts, S-phase-specific cyclin Cdk1 complex as well as Dfb4-Cdc7 kinase phosphorylate the initiation factors Sld2 and Sld3. These factors form a ternary complex with another initiation factor Dbp11 in their phosphorylated state, and associate with the origin of replication. This complex mediates the loading of Cdc45. A second complex called GINS and consisting of Sld5 and Psf1, 2 and 3 is also loaded onto the origin during the initiation process, in an interdependent manner with the Sld2/Sld3/Dpb11 complex. Both complexes cooperate in the recruitment of the replicative DNA polymerases, thus executing the initiation and subsequent establishment of the replication fork. Cdc45 and GINS are essential, well-conserved factors that are retained at the elongating replication fork. They form a stable helicase complex with MCM2-7 and mediate its contact to the replicative DNA polymerases. In contrast, the Sld2/Sld3/Dpb11 complex critical for the initiation is not retained by the elongating replication fork. Sld2 displays limited homology to the amino-terminal region of RecQL4 helicase, which may represent its metazoan orthologue, whereas Sld3 homologues have been identified only in fungi. Dbp11 and its fission yeast homologue Cut5 are members of a large family of BRCT-containing proteins including human TopBP1 and fruit fly Mus101. Similar principles of regulation apply also to human initiation of DNA replication, despite obvious differences in the detailed mechanisms. The regulatory initiation cascade is intimately intertwined with the cell cycle apparatus as well as the checkpoint control. PMID- 20012579 TI - Non-coding RNAs: new players in the field of eukaryotic DNA replication. AB - The machinery required for the replication of eukaryotic chromosomal DNA is made up of proteins whose function, structure and main interaction partners are evolutionarily conserved. Several new cases have been reported recently, however, in which non-coding RNAs play additional and specialised roles in the initiation of eukaryotic DNA replication in different classes of organisms. These non-coding RNAs include Y RNAs in vertebrate somatic cells, 26T RNA in somatic macronuclei of the ciliate Tetrahymena, and G-rich RNA in the Epstein-Barr DNA tumour virus and its human host cells. Here, I will give an overview of the experimental evidence in favour of roles for these non-coding RNAs in the regulation of eukaryotic DNA replication, and compare and contrast their biosynthesis and mechanisms of action. PMID- 20012580 TI - Function of TopBP1 in genome stability. AB - Human DNA topoisomerase IIbeta-binding protein 1 (TopBP1) and its orthologues in other organisms are proteins consisting of multiple BRCT modules that have acquired several functions during evolution. These proteins execute their tasks by interacting with a great variety of proteins involved in nuclear processes. TopBP1 is an essential protein that has numerous roles in the maintenance of the genomic integrity. In particular, it is required for the activation of ATM and Rad3-related (ATR), a vital regulator of DNA replication and replication stress response. The orthologues from yeast to human are involved in DNA replication and DNA damage response, while only proteins from higher eukaryotes are also involved in complex regulation of transcription, which is related to cell proliferation, damage response and apoptosis. We review here the recent progress in research aimed at elucidating the multiple cellular functions of TopBP1, focusing on metazoan systems. PMID- 20012581 TI - Eukaryotic single-stranded DNA binding proteins: central factors in genome stability. AB - The single-stranded DNA binding proteins (SSBs) are required to maintain the integrity of the genome in all organisms. Replication protein A (RPA) is a nuclear SSB protein found in all eukaryotes and is required for multiple processes in DNA metabolism such as DNA replication, DNA repair, DNA recombination, telomere maintenance and DNA damage signalling. RPA is a heterotrimeric complex, binds ssDNA with high affinity, and interacts specifically with multiple proteins to fulfil its function in eukaryotes. RPA is phosphorylated in a cell cycle and DNA damage-dependent manner with evidence suggesting that phosphorylation has an important function in modulating the cellular DNA damage response. Considering the DNA-binding properties of RPA a mechanism of "molecular counting" to initiate DNA damage-dependent signalling is discussed. Recently a human homologue to the RPA2 subunit, called RPA4, was discovered and RPA4 can substitute for RPA2 in the RPA complex resulting in an "alternative" RPA (aRPA), which can bind to ssDNA with similar affinity as canonical RPA. Additional human SSBs, hSSB1 and hSSB2, were recently identified, with hSSB1 being localized in the nucleus and having implications in DNA repair. Mitochondrial SSBs (mtSSBs) have been found in all eukaryotes studied. mtSSBs are related to prokaryotic SSBs and essential to main the genome stability in eukaryotic mitochondria. Recently human mtSSB was identified as a novel binding partner of p53 and that it is able to stimulate the intrinsic exonuclease activity of p53. These findings and recent results associated with mutations in RPA suggest a link of SSBs to cancer. PMID- 20012582 TI - DNA polymerases and mutagenesis in human cancers. AB - DNA polymerases (Pols) act as key players in DNA metabolism. These enzymes are the only biological macromolecules able to duplicate the genetic information stored in the DNA and are absolutely required every time this information has to be copied, as during DNA replication or during DNA repair, when lost or damaged DNA sequences have to be replaced with "original" or "correct" copies. In each DNA repair pathway one or more specific Pols are required. A feature of mammalian DNA repair pathways is their redundancy. The failure of one of these pathways can be compensated by another one. However, several DNA lesions require a specific repair pathway for error free repair. In many tumors one or more DNA repair pathways are affected, leading to error prone repair of some kind of lesions by alternatives routes, thus leading to accumulation of mutations and contributing to genomic instability, a common feature of cancer cell. In this chapter, we present the role of each Pol in genome maintenance and highlight the connections between the malfunctioning of these enzymes and cancer progress. PMID- 20012583 TI - DNA polymerase eta, a key protein in translesion synthesis in human cells. AB - Genomic DNA is constantly damaged by exposure to exogenous and endogenous agents. Bulky adducts such as UV-induced cyclobutane pyrimidine dimers (CPDs) in the template DNA present a barrier to DNA synthesis by the major eukaryotic replicative polymerases including DNA polymerase delta. Translesion synthesis (TLS) carried out by specialized DNA polymerases is an evolutionarily conserved mechanism of DNA damage tolerance. The Y family of DNA polymerases, including DNA polymerase eta (Pol eta), the subject of this chapter, play a key role in TLS. Mutations in the human POLH gene encoding Pol eta underlie the genetic disease xeroderma pigmentosum variant (XPV), characterized by sun sensitivity, elevated incidence of skin cancer, and at the cellular level, by delayed replication and hypermutability after UV-irradiation. Pol eta is a low fidelity enzyme when copying undamaged DNA, but can carry out error-free TLS at sites of UV-induced dithymine CPDs. The active site of Pol eta has an open conformation that can accommodate CPDs, as well as cisplatin-induced intrastrand DNA crosslinks. Pol eta is recruited to sites of replication arrest in a tightly regulated process through interaction with PCNA. Pol eta-deficient cells show strong activation of downstream DNA damage responses including ATR signaling, and accumulate strand breaks as a result of replication fork collapse. Thus, Pol eta plays an important role in preventing genome instability after UV- and cisplatin-induced DNA damage. Inhibition of DNA damage tolerance pathways in tumors might also represent an approach to potentiate the effects of DNA damaging agents such as cisplatin. PMID- 20012585 TI - Centromeres: assembling and propagating epigenetic function. AB - The faithful replication of DNA and the accurate segregation of genomic material from one generation to the next is critical in the maintenance of genomic stability. This chapter will describe the structure and assembly of an epigenetically inherited locus, the centromere, and its role in the processes by which sister chromatids are evenly segregated to daughter cells. During the G2 phase of the cell cycle kinetochores are assembled upon the chromatids. During mitosis, kinetochores attach chromosome(s) to the mitotic spindle. The kinetochore structure serves as the interface between the mitotic spindle and the chromatids and it is at the kinetochore where the forces that drive chromatid separation are generated. Unattached chromosomes fail to satisfy the spindle assembly checkpoint (SAC), resulting in cell cycle arrest. The centromere is the locus upon which the kinetochore assembles, and centromeres themselves are determined by their unique protein composition. Apart from budding yeast, centromeres are not specified simply by DNA sequence, but rather through chromatin composition and architecture and are thus epigenetically determined. Centromeres are built on a specific nucleosome not found elsewhere in the genome, in which histone H3 is replaced with a homologue - CENP-A or CenH3. This domain is flanked by heterochromatin and is folded to provide a 3-dimensional cylinder like structure at metaphase that establishes the kinetochore on the surface of the mitotic chromosomes. A large family of CENtromere Proteins (CENPs) associates with centromeric chromatin throughout the cell cycle and are required for kinetochore function. Unlike the bulk of histones, CENP-A is not assembled concurrently with DNA synthesis in S-phase but rather assembles into the centromere in the subsequent G1 phase. The assembly of CENP-A chromatin following DNA replication and the re-establishment of this network of constitutive proteins have emerged as critical mechanisms for understanding how the centromere is replicated during the cell cycle. PMID- 20012586 TI - Nucleotide excision repair in higher eukaryotes: mechanism of primary damage recognition in global genome repair. AB - Nucleotide excision repair (NER) is one of the major DNA repair pathways in eukaryotic cells that counteract the formation of genetic damage. NER removes structurally diverse lesions such as pyrimidine dimers, arising upon UV irradiation, and bulky chemical adducts, arising upon exposure to carcinogens and some chemotherapeutic drugs. NER defects lead to severe diseases, including some forms of cancer. In view of the broad substrate specificity of NER, it is of interest to understand how a certain set of proteins recognizes various DNA lesions in the contest of a large excess of intact DNA. This review focuses on DNA damage recognition, the key and, as yet, most questionable step of NER. Understanding of mechanism of this step of NER may give a key contribution to study of similar processes of DNA damage recognition (base excision repair, mismatch repair) and regulation of assembly of various DNA repair machines. The major models of primary damage recognition and pre-incision complex assembly are considered. The model of a sequential loading of repair proteins on damaged DNA seems most reasonable in the light of the available data. The possible contribution of affinity labeling technique in study of this process is discussed. PMID- 20012584 TI - The mitochondrial DNA polymerase in health and disease. AB - Since mutations in mitochondrial DNA (mtDNA) have been shown to be a cause of many mitochondrial diseases as well as aging, it is important to understand the origin of these mutations and how replication proteins modulate this process. DNA polymerase gamma (pol gamma) is the polymerase that is responsible for replication and repair of mtDNA. Pol gamma has three main roles in mtDNA maintenance and mutagenesis. As the only known DNA polymerase in mitochondria, pol gamma is required for all replication and repair functions and is the main source of errors produced in human mtDNA. Pol gamma is also sensitive to a host of antiviral nucleoside analogs used to treat HIV-1 infections, which can cause an induced mitochondrial toxicity. Finally, the gene for pol gamma, POLG, is a genetic locus for several mitochondrial disease with over 150 genetic mutations currently identified. PMID- 20012588 TI - Fluorescence-based quantification of pathway-specific DNA double-strand break repair activities: a powerful method for the analysis of genome destabilizing mechanisms. AB - This chapter provides instructions for the application of a fluorescence-based assay to examine different DNA double-strand break (DSB) repair pathways in primary mouse embryo fibroblasts (MEFs). The assay relies on targeted DSB formation in one of a series of repair substrates and subsequent repair-mediated reconstitution of the EGFP reporter. We present protocols for efficient introduction of extra-chromosomal repair substrate together with I-SceI endonuclease expression vector and subsequent measurement of DSB repair events down to frequencies of 0.001%. Concomitant transfection of plasmid and siRNA enables assessment of DSB repair under conditions of knockdown of protein expression, allowing to evaluate the contribution of single factors. Since the proteins of interest frequently have dual roles in DSB repair surveillance and checkpoint control, our assay procedure concomitantly corrects for transfection efficiencies, growth-, death-, and expression-related changes and also integrates the examination of the cell cycle status. PMID- 20012587 TI - Nonhomologous DNA end joining (NHEJ) and chromosomal translocations in humans. AB - Double-strand breaks (DSBs) arise in dividing cells about ten times per cell per day. Causes include replication across a nick, free radicals of oxidative metabolism, ionizing radiation, and inadvertent action by enzymes of DNA metabolism (such as failures of type II topoisomerases or cleavage by recombinases at off-target sites). There are two major double-strand break repair pathways. Homologous recombination (HR) can repair double-strand breaks, but only during S phase and typically only if there are hundreds of base pairs of homology. The more commonly used pathway is nonhomologous DNA end joining, abbreviated NHEJ. NHEJ can repair a DSB at any time during the cell cycle and does not require any homology, although a few nucleotides of terminal microhomology are often utilized by the NHEJ enzymes, if present. The proteins and enzymes of NHEJ include Ku, DNA-PKcs, Artemis, DNA polymerase mu (Pol micro), DNA polymerase lambda (Pol lambda), XLF (also called Cernunnos), XRCC4, and DNA ligase IV. These enzymes constitute what some call the classical NHEJ pathway, and in wild type cells, the vast majority of joining events appear to proceed using these components. NHEJ is present in many prokaryotes, as well as all eukaryotes, and very similar mechanistic flexibility evolved both convergently and divergently. When two double-strand breaks occur on different chromosomes, then the rejoining is almost always done by NHEJ. The causes of DSBs in lymphomas most often involve the RAG or AID enzymes that function in the specialized processes of antigen receptor gene rearrangement. PMID- 20012589 TI - Apoptosis: a way to maintain healthy individuals. AB - Apoptosis, the best known form of programmed cell death, is tightly regulated by a number of sensors, signal transducers and effectors. Apoptosis is mainly active during embryonic development, when deletion of redundant cellular material is required for the correct morphogenesis of tissues and organs; moreover, it is essential for the maintenance of tissue homeostasis during cell life. Cells also activate apoptosis when they suffer from various insults, such as damage to DNA or to other cellular components, or impairment of basic processes, such as DNA replication and DNA repair. Removal of damaged cells is fundamental in maintaining the health of organisms. In addition, apoptosis induction following DNA damage is exploited to kill cancer cells. In this chapter we will review the main features of developmental and induced apoptosis. PMID- 20012590 TI - The use of transgenic mice in cancer and genome stability research. AB - The development of effective cancer therapeutics is an important goal of modern biomedical sciences. To identify potential cancer therapeutic targets, the processes involved in tumorigenesis must be understood at all levels, which requires the development of model systems accurately mimicing tumor development. Cancer is the general name given to a variety of complex diseases characterised by uncontrolled cell proliferation. Cancer development is dependent not only on the changes occurring within the transformed cells, but also on the interactions of the cells with their microenvironment. The majority of our current understanding of carcinogenesis comes from the in vitro analysis of late-stage tumor tissue removed from cancer patients. While this has elucidated many genomic changes experienced by cancer cells, it provides little information about the factors influencing early-stage cancer development in vivo. Also certain hallmarks of cancer, such as metastasis and angiogenesis, are impossible to study in vitro. The mouse has become an important model for studying the in vivo aspects of human cancer development. Transgenic mouse models have been engineered to develop cancers, which accurately mimic their human counterparts, and have potential applications to test the effectiveness of novel cancer therapeutics. One of the most promising transgenic mouse models of human cancer arises from mice engineered with genomic instability. These transgenic models have been shown to develop human-like cancers and have the potential to provide insights into the molecular events occurring in earliest stages of tumorigenesis in vivo. PMID- 20012591 TI - [Ethical aspects of human embryonic stem cell use and commercial umbilical cord blood stem cell banking. Ethical reflections on the occasion of the regulation of the European Council and Parliament on advanced therapy medicinal products]. AB - The regulation of the European Council and Parliament on advanced therapy medicinal products also includes therapies with human embryonic stem cells. The use of these stem cells is controversially and heavily discussed. Contrary to the use of adult stem cells, medical and ethical problems concerning the use of human embryonic stem cells persists, because this use is based on the destruction of human life at the very beginning. The regulation foresees, therefore, subsidiarity within the European Member States. Although there are no ethical problems in principle with the use of stem cells from the umbilical cord blood, there are social ethical doubts with the banking of these stem cells for autologous use without any currently foreseeable medical advantage by commercial blood banks. Also in this case subsidiarity is valid. PMID- 20012592 TI - [Cervical paravertebral lesion]. AB - We present the case of a 56-year old male patient complaining of occasional shoulder pain and paresthesia in the left hand. Computer tomography and magnetic resonance imaging showed a left cervical lesion. Angiography was normal. Due to anatomical restrictions, the tumor could only be partly excised. Histologic examination showed a chordoma. The diagnosis and therapy options are presented and discussed. PMID- 20012593 TI - The rise of a ribosomopathy and increased cancer risk. PMID- 20012594 TI - Autonomic dysregulation in ob/ob mice is improved by inhibition of angiotensin converting enzyme. AB - The leptin-deficient ob/ob mice are insulin resistant and obese. However, the control of blood pressure in this model is not well defined. The goal of this study was to evaluate the role of leptin and of the renin-angiotensin system in the cardiovascular abnormalities observed in obesity using a model lacking leptin. To this purpose, we measured blood pressure in ob/ob and control animals by radiotelemetry combined with fast Fourier transformation before and after both leptin and enalapril treatment. Autonomic function was assessed pharmacologically. Blood pressure during daytime was slightly higher in the ob/ob compared to control mice, while no difference in heart rate was observed. Blood pressure response to trimetaphane and heart rate response to metoprolol were greater in ob/ob mice than in control littermates indicating an activated sympathetic nervous system. Heart rate response to atropine was attenuated. Baroreflex sensitivity and heart rate variability were blunted in ob/ob mice, while low frequency of systolic blood pressure variability was found increased. Chronic leptin replacement reduced blood pressure and reversed the impaired autonomic function observed in ob/ob mice. Inhibition of angiotensin-converting enzyme by enalapril treatment had similar effects, prior to the loss of weight. These findings suggest that the renin-angiotensin system is involved in the autonomic dysfunction caused by the lack of leptin in ob/ob mice and support a role of this interplay in the pathogenesis of obesity, hypertension, and metabolic syndrome. PMID- 20012596 TI - Pudendal nerve neuromodulation with neurophysiology guidance: a potential treatment option for refractory chronic pelvi-perineal pain. AB - Refractory chronic pelvi-perineal pain (RCPPP) is a challenging entity that has devastating consequences for patient's quality of life. Many etiologies have been proposed including pudendal neuralgia. Multiple treatment options are used but the reported results are sub-optimal and temporary. In this article, we present the technique of pudendal nerve neuromodulation with neurophysiology guidance as a treatment option for RCPPP. This technique is a two-step procedure that includes electrode implantation under neurophysiology guidance followed by the implantation of a permanent generator after a successful trial period. We report the cases of three women who underwent this procedure as a last-resort treatment option. After 2 years of follow-up, their symptoms are still significantly improved. No major complication occurred. PMID- 20012595 TI - Human ATP synthase beta is phosphorylated at multiple sites and shows abnormal phosphorylation at specific sites in insulin-resistant muscle. AB - AIMS/HYPOTHESIS: Insulin resistance in skeletal muscle is linked to mitochondrial dysfunction in obesity and type 2 diabetes. Emerging evidence indicates that reversible phosphorylation regulates oxidative phosphorylation (OxPhos) proteins. The aim of this study was to identify and quantify site-specific phosphorylation of the catalytic beta subunit of ATP synthase (ATPsyn-beta) and determine protein abundance of ATPsyn-beta and other OxPhos components in skeletal muscle from healthy and insulin-resistant individuals. METHODS: Skeletal muscle biopsies were obtained from lean, healthy, obese, non-diabetic and type 2 diabetic volunteers (each group n = 10) for immunoblotting of proteins, and hypothesis-driven identification and quantification of phosphorylation sites on ATPsyn-beta using targeted nanospray tandem mass spectrometry. Volunteers were metabolically characterised by euglycaemic-hyperinsulinaemic clamps. RESULTS: Seven phosphorylation sites were identified on ATPsyn-beta purified from human skeletal muscle. Obese individuals with and without type 2 diabetes were characterised by impaired insulin-stimulated glucose disposal rates, and showed a approximately 30% higher phosphorylation of ATPsyn-beta at Tyr361 and Thr213 (within the nucleotide-binding region of ATP synthase) as well as a coordinated downregulation of ATPsyn-beta protein and other OxPhos components. Insulin increased Tyr361 phosphorylation of ATPsyn-beta by approximately 50% in lean and healthy, but not insulin-resistant, individuals. CONCLUSIONS/INTERPRETATION: These data demonstrate that ATPsyn-beta is phosphorylated at multiple sites in human skeletal muscle, and suggest that abnormal site-specific phosphorylation of ATPsyn-beta together with reduced content of OxPhos proteins contributes to mitochondrial dysfunction in insulin resistance. Further characterisation of phosphorylation of ATPsyn-beta may offer novel targets of treatment in human diseases with mitochondrial dysfunction, such as diabetes. PMID- 20012598 TI - Prepubertal exposure to commercial formulation of the herbicide glyphosate alters testosterone levels and testicular morphology. AB - Glyphosate is a herbicide widely used to kill weeds both in agricultural and non agricultural landscapes. Its reproductive toxicity is related to the inhibition of a StAR protein and an aromatase enzyme, which causes an in vitro reduction in testosterone and estradiol synthesis. Studies in vivo about this herbicide effects in prepubertal Wistar rats reproductive development were not performed at this moment. Evaluations included the progression of puberty, body development, the hormonal production of testosterone, estradiol and corticosterone, and the morphology of the testis. Results showed that the herbicide (1) significantly changed the progression of puberty in a dose-dependent manner; (2) reduced the testosterone production, in semineferous tubules' morphology, decreased significantly the epithelium height (P < 0.001; control = 85.8 +/- 2.8 microm; 5 mg/kg = 71.9 +/- 5.3 microm; 50 mg/kg = 69.1 +/- 1.7 microm; 250 mg/kg = 65.2 +/- 1.3 microm) and increased the luminal diameter (P < 0.01; control = 94.0 +/- 5.7 microm; 5 mg/kg = 116.6 +/- 6.6 microm; 50 mg/kg = 114.3 +/- 3.1 microm; 250 mg/kg = 130.3 +/- 4.8 microm); (4) no difference in tubular diameter was observed; and (5) relative to the controls, no differences in serum corticosterone or estradiol levels were detected, but the concentrations of testosterone serum were lower in all treated groups (P < 0.001; control = 154.5 +/- 12.9 ng/dL; 5 mg/kg = 108.6 +/- 19.6 ng/dL; 50 mg/dL = 84.5 +/- 12.2 ng/dL; 250 mg/kg = 76.9 +/- 14.2 ng/dL). These results suggest that commercial formulation of glyphosate is a potent endocrine disruptor in vivo, causing disturbances in the reproductive development of rats when the exposure was performed during the puberty period. PMID- 20012600 TI - Fatigue related changes in electromyographic coherence between synergistic hand muscles. AB - The aim of this study was to examine coherence between surface electromyographic (EMG) signals from two index finger flexor muscles, the first dorsal interosseous (FDI) and flexor digitorum superficialis (FDS), during and immediately following sustained, fatiguing isometric contraction. Coherence was observed between the FDI and FDS EMG signals in the tremor (8-12 Hz), beta (15-35 Hz) and gamma (35-60 Hz) bands in all subjects. A significant increase in EMG-EMG coherence in the beta and gamma frequency bands was observed immediately following the fatiguing contraction. No significant difference was observed in the tremor band coherence before and after fatigue. Coherence was observed between EMG and force in the tremor band during both the pre- and post-fatigue contractions and a significant increase in the FDI EMG-force coherence post-fatigue was observed. It is suggested that the increase in beta and gamma band coherence with fatigue may be due to increased levels of corticomotoneuronal drive to both muscles. Alternatively, the increased EMG-EMG coherence may reflect an increased contribution of peripheral afferents to coupling across the muscle with fatigue. Although the functional significance is not clear, the increase in coherence may help to overcome reduced motoneuron excitability with fatigue, to bind together different sensorimotor elements or to coordinate force generation across muscles in a more synergistic manner as the force generating capacity of the muscle is decreased. PMID- 20012599 TI - Antipointing: perception-based visual information renders an offline mode of control. AB - Recent work by our group reported that mirror-symmetrical reaching movements (i.e., antipointing) are supported by perception-based visual information (Heath et al. 2009a). The present study was designed to determine if the perception based visual information supporting antipointing results in a primarily offline mode of control; that is, a mode of control wherein the response unfolds with few if any-online limb corrections. Participants reached directly to (propointing) or mirror-symmetrical (antipointing) to targets presented in the left and right visual fields. To examine the extent reaches were controlled online versus offline, we computed the proportion of variance (R (2)) explained by the position of the limb at 75% of movement time relative to the response's ultimate movement endpoint. The endpoints for propointing in left and right visual fields demonstrated robust endpoint accuracy and stability and were tied to low R (2) values. In contrast, antipointing elicited a marked degree of endpoint variability and were characterized by a visual-field specific pattern of endpoint bias. Moreover, the R (2) values for antipointing were more robust than propointing counterparts. Therefore, we propose that the metrical visual information supporting stimulus-driven propointing results in a primarily online mode of control whereas the relative nature of the perception-based visual information supporting antipointing results in an offline mode of control. More directly, we propose that the offline control of antipointing is attributed to the "slow" cognitive processing of visuoperceptual networks. PMID- 20012601 TI - Pharmacokinetic interaction between oral lovastatin and verapamil in healthy subjects: role of P-glycoprotein inhibition by lovastatin. AB - BACKGROUND: Lovastatin is an inhibitor of P-glycoprotein (P-gp) and is metabolized by the cytochrome P450 (CYP) 3A4 isoenzyme. Verapamil is a substrate of both P-gp and CYP3A4. It is therefore likely that lovastatin can alter the absorption and metabolism of verapamil. METHODS: The pharmacokinetic parameters of verapamil and one of its metabolites, norverapamil, were compared in 14 healthy male Korean volunteers (age range 22-28 years) who had been administered verapamil (60 mg) orally in the presence or absence of oral lovastatin (20 mg). The design of the experiment was a standard 2 x 2 crossover model in random order. RESULTS: The pharmacokinetic parameters of verapamil were significantly altered by the co-administration of lovastatin compared to the control. The area under the plasma concentration-time curve (AUC (0-infinity)) and the peak plasma concentration of verapamil were significantly increased by 62.8 and 32.1%, respectively. Consequently, the relative bioavailability of verapamil was also significantly increased (by 76.5%). The (AUC (0-infinity)) of norverapamil and the terminal half-life of verapamil did not significantly changed with lovastatin coadministration. The metabolite-parent ratio was significantly reduced (29.2%) in the presence of lovastatin. CONCLUSION: Lovastatin increased the absorption of verapamil by inhibiting P-gp and inhibited the first-pass metabolism of verapamil by inhibiting CYP3A4 in the intestine and/or liver in humans. PMID- 20012602 TI - Functional analysis in MR urography - made simple. AB - MR urography (MRU) has proved to be a most advantageous imaging modality of the urinary tract in children, providing one-stop comprehensive morphological and functional information, without the utilization of ionizing radiation. The functional analysis of the MRU scan still requires external post-processing using relatively complex software. This has proved to be a limiting factor in widespread routine implementation of MRU functional analysis and use of MRU functional parameters similar to nuclear medicine. We present software, developed in a pediatric radiology department, that not only enables comprehensive automated functional analysis, but is also very user-friendly, fast, easily operated by the average radiologist or MR technician and freely downloadable at www.chop-fmru.com . A copy of IDL Virtual Machine is required for the installation, which is obtained at no charge at www.ittvis.com . The analysis software, known as "CHOP-fMRU," has the potential to help overcome the obstacles to widespread use of functional MRU in children. PMID- 20012603 TI - Developmental and tissue-specific expression of NITRs. AB - Novel immune-type receptors (NITRs) are encoded by large multi-gene families and share structural and signaling similarities to mammalian natural killer receptors (NKRs). NITRs have been identified in multiple bony fish species, including zebrafish, and may be restricted to this large taxonomic group. Thirty-nine NITR genes that can be classified into 14 families are encoded on zebrafish chromosomes 7 and 14. Herein, we demonstrate the expression of multiple NITR genes in the zebrafish ovary and during embryogenesis. All 14 families of zebrafish NITRs are expressed in hematopoietic kidney, spleen and intestine as are immunoglobulin and T cell antigen receptors. Furthermore, all 14 families of NITRs are shown to be expressed in the lymphocyte lineage, but not in the myeloid lineage, consistent with the hypothesis that NITRs function as NKRs. Sequence analyses of NITR amplicons identify known alleles and reveal additional alleles within the nitr1, nitr2, nitr3, and nitr5 families, reflecting the recent evolution of this gene family. PMID- 20012604 TI - Naturally occurring systemic immune responses to HPV antigens do not predict regression of CIN2/3. AB - Essentially all squamous cervical cancers and their precursor lesions, high grade cervical intraepithelial neoplasia (CIN2/3), are caused by persistent human papillomavirus (HPV) infection. However, not all CIN2/3 lesions progress to cancer. In a brief, observational study window monitoring subjects with CIN2/3 from protocol entry (biopsy diagnosis) to definitive therapy (cervical conization) at week 15, in a cohort of 50 subjects, we found that 26% of CIN2/3 lesions associated with HPV16, the genotype most commonly associated with disease, underwent complete histologic regression. Nonetheless, HPV16-specific T cell responses measured in peripheral blood obtained at the time of study entry and at the time of conization were marginally detectable directly ex vivo, and did not correlate with lesion regression. This finding suggests that, in the setting of natural infection, immune responses which are involved in elimination of cervical dysplastic epithelium are not represented to any great extent in the systemic circulation. PMID- 20012605 TI - Patients with oral squamous cell carcinoma are characterized by increased frequency of suppressive regulatory T cells in the blood and tumor microenvironment. AB - Oral squamous cell carcinoma (OSCC) is a cancerous lesion with high incidence worldwide. The immunoregulatory events leading to OSCC persistence remain to be elucidated. Our hypothesis is that regulatory T cells (Tregs) are important to obstruct antitumor immune responses in patients with OSCC. In the present study, we investigated the frequency, phenotype, and activity of Tregs from blood and lesions of patients with OSCC. Our data showed that >80% of CD4(+)CD25(+) T cells isolated from PBMC and tumor sites express FoxP3. Also, these cells express surface Treg markers, such as GITR, CD45RO, CD69, LAP, CTLA-4, CCR4, and IL-10. Purified CD4(+)CD25(+) T cells exhibited stronger suppressive activity inhibiting allogeneic T-cell proliferation and IFN-gamma production when compared with CD4(+)CD25(+) T cells isolated from healthy individuals. Interestingly, approximately 25% of CD4(+)CD25(-) T cells of PBMC from patients also expressed FoxP3 and, although these cells weakly suppress allogeneic T cells proliferative response, they inhibited IFN-gamma and induced IL-10 and TGF-beta secretion in these co-cultures. Thus, our data show that Treg cells are present in OSCC lesions and PBMC, and these cells appear to suppress immune responses both systemically and in the tumor microenvironment. PMID- 20012606 TI - Residual nodal disease in biopsy proven n1/n2 breast cancer following neoadjuvant systemic therapy. AB - BACKGROUND: The purpose of this study was to determine the rate of complete pathologic response in axillary lymph nodes after neoadjuvant therapy and the clinicopathologic factors associated with a complete response. METHODS: Clinical, demographic, and pathologic data from all patients with breast cancer treated at our institution are prospectively recorded in a database. We reviewed this database from 2000 to 2007 and identified 90 patients who were node-positive before neoadjuvant therapy based on image-guided fine needle aspiration biopsy; all 90 patients underwent axillary lymph node dissection (ALND) after neoadjuvant therapy. Data were compared using chi-square and Fisher's exact test. RESULTS: Of 90 patients with breast cancer who were node-positive before neoadjuvant therapy, 71 (79%) had positive nodal disease on final ALND pathology and 19 (21%) had a complete nodal pathologic response. Age, race, tumor grade, clinical T and N stage, and estrogen/progesterone receptor and Her-2neu status were not predictive of a complete nodal response. The only factor predictive of a complete nodal response was the type of neoadjuvant therapy used; all 19 patients with a complete response received neoadjuvant chemotherapy and none received neoadjuvant endocrine therapy (P < 0.05). CONCLUSIONS: Twenty-five percent of patients who underwent neoadjuvant chemotherapy had a complete pathologic response in the nodal basin, whereas no patient who underwent neoadjuvant endocrine therapy experienced a complete nodal response. Twenty-five percent of patients who underwent neoadjuvant chemotherapy had a complete pathological response in the nodal specimen, whereas no patient who underwent neoadjuvant endocrine therapy experienced a complete nodal response. PMID- 20012607 TI - Poor late survival after surgical treatment of pleural empyema. AB - BACKGROUND: We have evaluated the outcome of patients who underwent surgical treatment of parapneumonic pleural empyema. METHODS: This study included 143 consecutive patients older than 18 years who were operated on for parapneumonic pleural empyema. RESULTS: The overall survival at 30 days, 3 months, 5 years, and 10 years was 97.2, 92.3, 80.6, and 61.9%, respectively. Serum albumin (OR = 0.78, 95% CI = 0.65-0.94), cerebrovascular disease (OR = 30.49, 95% CI = 1.35-689.05), pulmonary embolism (OR = 984.63, 95% CI = 7.81-124206.8), and Thoracoscore (OR = 1.61, 95% CI = 1.10-2.35) were independent predictors of 3-month overall survival. Age (RR = 1.08, 95% CI = 1.03-1.14), serum albumin (RR = 0.89, 95% CI = 0.82-0.98), chronic obstructive pulmonary disease (COPD) (RR = 5.14, 95% CI = 1.3319.84), and cerebrovascular disease (RR = 6.76, 95% CI = 1.33-34.34) were independent predictors of pneumonia/pleural empyema-related death. Twenty-two patients required 33 reinterventions after the primary procedure: 19 patients after primary thoracoscopy and 3 patients after primary thoracotomy. Reintervention did not have any significant impact on 30-day mortality (4.5% vs. 2.5%, p = 0.49), but it affected 3-month mortality (26.1% vs. 4.2%, p < 0.0001). Reoperation was an independent predictor of late overall survival (at 10 years: 35.5% vs. 67.4%, RR = 2.95, 95% CI = 1.33-6.57) and freedom from pneumonia/pleural empyema-related death (at 10 years: 73.9% vs. 91.3%, RR = 4.40, 95% CI = 1.24-15.66). CONCLUSIONS: Surgical treatment of pleural empyema can be associated with good immediate results, but longer follow-up showed that pneumonia/pleural empyema-related mortality and all-cause mortality are rather poor. PMID- 20012608 TI - John Jones, M.D.: pioneer, patriot, and founder of American surgery. AB - John Jones was a pioneer of American Surgery. Born in Long Island, New York in 1729, he received his medical degree in France from the University of Rheims. He returned to the colonies and helped to establish the medical school that would later become Columbia University's College of Physicians and Surgeons where he was appointed the first Professor of Surgery in the New World. He used his position to assert that surgeons trained in America should be familiar with all facets of medicine and not be mere technicians. Before the outbreak of the American Revolution, he wrote a surgical field manual, which was the first medical text published in America. A believer in the principles of the American Revolution, he would go on to count Benjamin Franklin and George Washington as his patients. Despite achieving many firsts in American medicine, his influence on surgical training is his most enduring legacy. PMID- 20012609 TI - Left heart pump-assisted myocardial revascularization favorably affects neutrophil apoptosis. AB - OBJECTIVES: Granulocyte apoptosis is a key control process in the clearance of neutrophils from inflammatory sites, and its rate is modulated by a number of inflammatory mediators. In this study, we investigated whether the use of left ventricular-assisted technique (LVA) in beating heart myocardial revascularization would exert less impact on neutrophil apoptosis compared with conventional cardiopulmonary bypass (CPB). METHODS: Forty consecutive patients who underwent myocardial revascularization were randomly assigned to LVA (group A, 21 patients) or CPB (group B, 19 patients). Blood samples for detection of interleukin-6, interleukin-8, and tumor necrosis factor-alpha were measured at baseline and at various time points postoperatively. Neutrophil apoptosis was detected by light microscopy as well as by the annexin-V assays together with the activity of caspase 3 on postoperative samples. RESULTS: Preoperative clinical and demographic data did not differ between the two groups. The two groups also were similar with respect to mortality, number of grafts performed, duration of extracorporeal circulation, and need for inotropes. However postoperatively, spontaneous apoptosis was significantly delayed in neutrophils from CPB patients compared with LVA patients. Neutrophils were activated, as indicated by increased surface expression of CD11b. Caspase 3 activity was found to be significantly reduced in neutrophils from CPB patients after 18 and 24 hours of culture. CONCLUSIONS: Patients who underwent beating heart myocardial revascularization with LVA show a better preserved neutrophil apoptosis than patients treated with the CPB. PMID- 20012610 TI - Multimedia abstract generation of intensive care data: the automation of clinical processes through AI methodologies. AB - Medical errors from communication failures are enormous during the perioperative period of cardiac surgical patients. As caregivers change shifts or surgical patients change location within the hospital, key information is lost or misconstrued. After a baseline cognitive study of information need and caregiver workflow, we implemented an advanced clinical decision support tool of intelligent agents, medical logic modules, and text generators called the "Inference Engine" to summarize individual patient's raw medical data elements into procedural milestones, illness severity, and care therapies. The system generates two displays: 1) the continuum of care, multimedia abstract generation of intensive care data (MAGIC)-an expert system that would automatically generate a physician briefing of a cardiac patient's operative course in a multimodal format; and 2) the isolated point in time, "Inference Engine"-a system that provides a real-time, high-level, summarized depiction of a patient's clinical status. In our studies, system accuracy and efficacy was judged against clinician performance in the workplace. To test the automated physician briefing, "MAGIC," the patient's intraoperative course, was reviewed in the intensive care unit before patient arrival. It was then judged against the actual physician briefing and that given in a cohort of patients where the system was not used. To test the real-time representation of the patient's clinical status, system inferences were judged against clinician decisions. Changes in workflow and situational awareness were assessed by questionnaires and process evaluation. MAGIC provides 200% more information, twice the accuracy, and enhances situational awareness. This study demonstrates that the automation of clinical processes through AI methodologies yields positive results. PMID- 20012611 TI - Evaluation of the adequacy of lymph node dissection in pylorus-preserving gastrectomy for early gastric cancer using the maruyama index. AB - BACKGROUND: The Maruyama computer program predicts the percentage likelihood of disease in lymph node stations left undissected by a surgeon, according to the age and sex of the patient and the gross type, size, location, depth of invasion, and histology of the tumor. The Maruyama index (MI) is defined as the sum of the predictions of the percentage likelihood of disease in undissected regional lymph node station (station 1-12). It has been shown that an MI < 5 is a strong predictor of survival and that the MI is an independent predictor of overall survival and relapse risk. We used the MI to evaluate the adequacy of lymph nodes dissection in pylorus-preserving gastrectomy (PPG) for early gastric cancer (EGC) performed at Seoul National University Hospital. METHODS: From March 2003 to September 2007, PPG was performed for patients with EGC of the middle third of the stomach when the distal resection margin was greater than 2 cm and preservation of 3 cm antral segment was possible. MIs and pathologic data such as TNM stage and the presence of metastatic lymph node for each station were reviewed. RESULTS: PPG were performed on 24 patients. The mean age of the patients was 55 years. The median tumor size was 2.1 cm. The median and mean MI were 0 and 0.8, respectively. There were 23 cases of T1N0M0 tumor and one case of T2N0M0 tumor postoperatively. There were no recurrences (mean follow-up period: 27.2 months). CONCLUSIONS: The MI indicates that lymph node dissection during PPG performed in these 24 patients was adequate. PMID- 20012612 TI - Effect of liver transplantation on spleen size, collateral veins, and platelet counts. AB - BACKGROUND: The aim of this study was to evaluate the effect of liver transplantation on the spleen size, spontaneous splenorenal shunt (SRS) function, and platelet counts in patients with hypersplenism. METHODS: Between December 2001 and February 2007, 462 adult patients underwent orthotopic liver transplantations (OLTX) at our institution. Of these patients, CT or MRI information was reviewed retrospectively in 55 patients. Volume measurements of the spleen and liver, spleen/liver volume ratio (S/L ratio), presence and size of SRS, and platelet counts were evaluated before and after OLTX. RESULTS: Mean spleen volume decreased from 827 +/- 463 ml to 662 +/- 376 ml after OLTX (p < 0.01). Five (11%) patients returned to normal-range spleen size after OLTX. SRS was observed in 19 patients before OLTX (35%). The diameter of SRS also significantly decreased from 1.0 +/- 0.5 cm before OLTX to 0.7 +/- 0.5 cm after OLTX (p < 0.05). SRS disappeared in 16% of patients (3/19). S/L ratio significantly decreased from 0.65 +/- 0.33 to 0.38 +/- 0.17 (p < 0.01) after OLTX. Platelet counts significantly increased after OLTX (p < 0.01). Improvement of the platelet count in the group with postoperative S/L ratio >0.35 was not as good as that in the group with S/L ratio <0.35 (p < 0.01). CONCLUSIONS: Spleen size and SRS size became significantly smaller after OLTX. However, patients with postoperative S/L ratio >0.35 tend to have lower platelet counts after OLTX. PMID- 20012613 TI - Application of a visible simulation surgery technique in preoperation planning for intrahepatic calculi. AB - BACKGROUND: The aim of this study was to evaluate a visible simulation surgery technique for choosing the best surgical plan in patients with intrahepatic calculi. METHODS: A medical image processing system was used to process computed tomography (CT) scanning data collected from four cases of intrahepatic calculi. Models of liver and bile ducts in standard template library format were processed by a free-form modeling system and reconstructed three-dimensionally. Accurate digital information about the bile duct system, lesions, calculi distribution, and adjacent organs from all directions, multiple angles, and multi-strata were used to choose the best surgical plan. Then, visible simulation surgery was performed with simulation operation software. RESULTS: Three-dimensionally reconstructed models provide clarity with strong relief perception and a user friendly interface. Visible simulation surgery performed based on three dimensionally reconstructed models led to optimal operation planning. CONCLUSIONS: Visible simulation surgery is more objective and complete than routine preoperative examinations to choose the best operation plan for intrahepatic calculi. PMID- 20012614 TI - Performance of a computerized protocol for trauma shock resuscitation. AB - BACKGROUND: A computerized protocol was developed and used to standardize bedside clinician decision making for resuscitation of shock due to severe trauma during the first day in the intensive care unit (ICU) at a metropolitan Level I trauma center. We report overall performance of a computerized protocol for resuscitation of shock due to severe trauma, incorporating two options for resuscitation monitoring and intervention intensity, according to: (1) duration of use and (2) acceptance of computerized protocol-generated instructions. METHODS: A computerized protocol operated by clinicians, using a personal computer (PC) at the bedside, was used to guide clinical decision making for resuscitation of patients meeting specific injury and shock criteria. The protocol generated instructions that could be accepted or declined. Clinician acceptance of the protocol instructions was stored by the PC software in a database for each patient. A rule-based, data-driven protocol was developed using literature evidence, expert opinion, and ongoing protocol performance analysis. Logic-flow diagrams were used to facilitate communication among multidisciplinary protocol development team members. The protocol was computerized using standard programming methods and implemented using cart-mounted PCs with a touch screen and keyboard interfaces. Protocol progression began with patient demographic data and criteria entry, confirmation of hemodynamic monitor instrumentation, request for specific hemodynamic performance data, and instructions for specific interventions (or no intervention). Use and performance of the computerized protocol was recorded in a protocol execution database. The protocol was continuously maintained with new literature evidence and database performance analysis findings. Initially implemented in 2000, the computerized protocol was refined in 2004 with two options for resuscitation intensity: pulmonary artery catheter- and central venous pressure-directed resuscitation. RESULTS: Over 2 years ending at August 2006, a total of 193 trauma patients (mean Injury Severity Score was 27, survival rate 89%) were resuscitated using the computerized protocol. Protocol duration was 4400 hours or 22.7 +/- 0.4 hours per patient. The computerized protocol generated 3724 instructions (19 +/- 1 per patient) that required a bedside clinician response. In all, 94% of these instructions were accepted by the bedside clinician users. CONCLUSIONS: A computerized protocol to guide decision making for trauma shock resuscitation in a Level 1 trauma center surgical ICU was developed and used as standard of care. During 2 years ending at August 2006, 94% of computer-generated instructions for specific interventions or measurements of hemodynamic performance were accepted by bedside clinicians, indicating appropriate, useful design and reliance on the computerized protocol system. PMID- 20012615 TI - Residual skin tags following procedure for prolapse and hemorrhoids: differentiation from recurrence. AB - PURPOSE: The aim of this study was to compare the incidence of recurrence and residual skin tag after the procedure for prolapse and hemorrhoids (PPH) versus conventional hemorrhoidectomy (CH) in the Chinese- and English-language literature to explore the definition of recurrence after PPH. METHODS: Related Chinese- and English-language literature was collected by several methods. Meta analysis was used to compare the incidence of recurrence and residual skin tag of PPH versus CH. RESULTS: In China, 13.94% of hemorrhoids had a skin tag after PPH. The mean recurrence rate after PPH was 3.23% (range 0.40-26.44%). Our meta analysis of PPH versus CH of the Chinese studies showed that PPH had a significantly lower recurrence rate-13 studies, odds ratio (OR) 0.27, 95% confidence interval (CI) 0.17-0.42, p < 0.00001-and a higher incidence of skin tags with no significance-6 studies, OR 3.42, 95% CI 0.49-24.04, p = 0.22. Our meta-analysis of PPH versus CH among the English-language studies showed that PPH had a significantly higher recurrence rate-17 studies, 636 patients in the PPH group vs. 625 patients in the CH group, OR 2.96, 95% CI 1.57-5.56, p = 0.0008-and a significantly higher incidence of residual skin tags-8 studies, 297 patients in the PPH group vs. 289 patients in the CH group, OR 1.88, 95% CI 1.15-3.05, p = 0.01. However, the recurrence of prolapse was stated to be ascertained by anorectal examination in only six studies; meta-analysis of the six studies showed that PPH was not associated with a higher recurrence-six studies, 230 patients in the PPH group vs. 220 patients in the CH group; OR 1.87, 95% CI 0.70 5.00, p = 0.22. CONCLUSIONS: PPH is not associated with a higher recurrence rate but is associated with a higher incidence of skin tags compared with CH. The reported high recurrence rates are probably caused by improper inclusion of residual skin tags into the recurrence data. Surgeons should perform anorectal examinations to differentiate a residual skin tag from a recurrence. PMID- 20012616 TI - A comparison of carotid surgery in northern Europe and northern Africa. AB - BACKGROUND: Although the indications for carotid endarterectomy (CEA) are clearly defined by major trials, CEA has not been properly implemented in many developing regions, including Northern Africa represented in this study by Upper Egypt. This study was designed to estimate the need for CEA in symptomatic patients with significant internal carotid artery stenosis in Upper Egypt. The estimated needs and actual provision of CEA in Upper Egypt were compared with those of Uusimaa (Finland) and Wessex (England) regions, representing Northern Europe. METHODS: Incidence rates were derived from a community-based door-to-door survey in Upper Egypt, Oxford Community Stroke Project, and epidemiological Finnish studies. The provision of CEA was derived from the local registry of Vascular Surgery Department, Assiut University Hospitals in Upper Egypt; HUSVASC data registry at Helsinki University Central Hospital, and the published data of the Wessex region. The estimated needs were calculated using previously published proportions of patients eligible for CEA. RESULTS: The population at risk of cerebral ischemic events (>or=65 years old) is 4% of the Egyptian population compared with 16% and 15% of the British and Finnish populations, respectively. The incidence of stroke and transient ischemic attack (TIA) is comparable in England and Finland but higher than the Egyptian rates (1.9 per thousand and 2.1 per thousand vs. 1.2 per thousand for stroke; 0.5 per thousand and 0.6 per thousand vs. 0.2 per thousand for TIA, respectively). The ratio of the actual provision of CEA to the estimated need in Wessex and Uusimaa is the same (0.5), whereas it is much lower (0.003) in Upper Egypt. CONCLUSIONS: Despite the low incidence of stroke and TIA, there is a huge unmet need for CEA in Upper Egypt. Yet, Wessex and Uusimaa also seemed to be at a suboptimal level compared with the estimated need. If CEA could be offered annually to those 1,650 patients with severe symptomatic ICA stenosis, 275 strokes could be prevented in Upper Egypt in 5-year duration, provided that the operative morbidity and mortality rates are equal to those reported in the large, randomized, controlled trials. PMID- 20012617 TI - The thin sectional anatomy of the sellar region with MRI correlation. AB - The purpose of this study was to explore the anatomical complexity and adjacent relationships of the sellar region in thin continuous sections so as to provide intimate morphological data for imaging diagnosis and surgical operations of the diseases in this region. After CT and MR examination verifying no brain lesions, one normal cadaver head was selected for this study from four Chinese adult male cadavers. After being embedded and frozen, the head was sliced into serial sections at 0.1 mm intervals in the transverse plane with SKC 500 computerized freezing milling machine. Then the serial transverse sections were photographed by a high-resolution digital camera and saved in the computer. Subsequently, the anatomic structures of the sellar region on the thin transverse sections were investigated and correlated with the MR images of the specimen as well as in vivo MR images, which were obtained from 20 normal Chinese male adult volunteers by a 3.0 T GE MR scanner. The base lines of the sectioning and the MR scan were all parallel to the AC-PC line. A total of 320 transverse sections and 10-12 transverse MR images related with the sellar region were obtained, respectively. We investigated the sectional anatomy of the sellar region and divided it into three parts: supra hypophysial area, hypophysial area and infra hypophysial area. The cavernous sinus was a venous passage full of blood and it could be divided into four interspaces according to its position relation with the internal carotid artery. The third, fourth, sixth cranial nerves and trigeminal branches ophthalmic nerve, maxillary nerve displayed from the anterior to the posterior in the lateral wall of cavernous sinus in transverse planes. Comparing continuous thin sections with MR images offers a better understanding of the complex anatomical structures and provides practical submillimeter anatomical data for imaging diagnosis and clinical treatment in this region. PMID- 20012618 TI - Study on inferior petrosal sinus and its confluence pattern with relevant veins by MSCT. AB - OBJECTIVE: To explore the anatomic route of inferior petrosal sinus (IPS) after going out of the cranium and its confluence patterns with internal jugular vein (IJV), anterior condylar vein (ACV) and lateral condylar vein (LCV), and to supply knowledge about typing of IPS-IJV junction, so as to provide reference evidence for evaluation of transvenous access route in the diagnosis and treatment of skull base and cavernous sinus lesions. METHODS: In 120 patients, the IPS route and its confluence with IJV, ACV and LCV were shown by multi-planar reconstruction (MPR) and curve multi-planar reconstruction (CMPR). Combining with continuous thin slice, the IPS-IJV junction was further subdivided according to the level of IPS confluence with IJV and whether there is an anastomosis with sigmoid sinus (SS). Furthermore, the IPS length, venous diameter at IPS-IJV junction and IPS-SS communicating branch were determined and compared. RESULTS: Inferior petrosal sinus directly draining into jugular bulb (JB) or/and draining into JB after communication with SS was found in 28 sides (11.7%, pattern A); IPS draining into IJV at the level of exterior opening of hypoglossal canal in 114 sides (47.5%, pattern B); IPS draining into IJV in a lower extracranial level in 83 sides (34.6%, pattern C); IPS with multiple junctions draining into IJV near the jugular foramen in 12 sides (5.0%, pattern D); IPS directly draining into VVP in 1 side (0.4%, pattern E); IPS being absent in 1 side (0.4%, pattern F). IPS draining into VVP via ACV was seen in 218 sides, IPS draining into VVP via LCV in 100 sides and IPS directly draining into IJV in 14 sides. After going out of the cranium, IPS goes along with IJV for a relatively long distance in some cases. The IPS extracranial length over 40 mm was found in ten sides and the lowest level of IPS route was at the inferior margin of the fourth cervical vertebra. The venous diameter at the IPS-IJV confluence was 0.8-5.7 mm (mean 2.51 mm) and it was significantly larger on the right side than on the left (P = 0.01). However, there was no remarkable difference between male patients and female ones. CONCLUSION: Continuous thin-slice scanning by multislice spiral computed tomography in combination with MPR and CMPR can clearly show IPS route and its confluence with relevant veins, and determine the feasibility of procedures via IPS. Therefore, it can be used as an effective method for preoperative evaluation of IPS for diagnosis and treatment of skull base and cavernous sinus lesions by IJV access route. PMID- 20012619 TI - Characteristics of biofilms from urinary tract catheters and presence of biofilm related components in Escherichia coli. AB - Long term catheterization of the urinary tract leads to bacterial colonization of the urine, whereby adherence to the catheter surface is a major determinative factor for colonization. Collection of bacterial isolates from urine and urinary catheters of 45 patients showed multi-species catheter-colonization, while Escherichia coli isolates were frequently found in the urine in high numbers. Biofilm formation of catheter and urine-derived E. coli isolates was associated with the presence of the fluA gene, loss of O-antigen, and expression of type 1 fimbriae. The second messenger cyclic di-GMP (cdiGMP), a major regulator of biofilm formation, regulated adherence to the catheter surface in a selected clinical isolate suggesting that the cdiGMP second messenger pathway may be a target for anti-biofilm therapeutic approaches. PMID- 20012620 TI - [Acetone compression. A fast, standardized method to investigate gastrointestinal lymph nodes]. AB - Lymph node staging is the most important prognostic parameter in malignant gastrointestinal tumors. Manual dissection of adipose tissue is time-consuming and also depends on the experience of the individual examiner. By combining elution with acetone and mechanical compression using simple equipment it was possible to completely embed adipose tissue from 404 surgical specimens (colon 348, stomach 28, greater omentum 14, other location 14) without manual dissection. As a result of the procedure, the weight of the adipose tissue could be reduced by 90%-95%, making full histological examination possible. The colon specimens included an average of 43.8 lymph nodes (14-109) in 14 embedding cassettes (1-38) with a native fat weight of 234.7 g (42.8-820 g). The quality of histological staining, including immunohistochemical and molecular investigations, is of comparable quality to routine work-up. Elution with acetone enables the prompt, standardized and full histological work-up of adipose tissue without manual dissection. Moreover, additional costs are low. The number of lymph nodes required by medical associations was attained in all cases and often exceeded. This method was successfully used in other organs (greater omentum, breast). PMID- 20012621 TI - Primary antiphospholipid syndrome with and without Sneddon's syndrome. AB - The main objective of this study was to compare clinical and laboratory data obtained from patients with primary antiphospholipid syndrome (PAPS) with and without Sneddon's syndrome (SS). A transverse study with 54 (85.2% female) PAPS patients (Sapporo criteria) was performed. Demographic, drug use, and antiphospholipid antibodies data were evaluated, as well as clinical and laboratory findings of SS. Patients were subdivided into one of two groups: PAPS with SS and PAPS without SS. Both groups were similar with respect to age (p = 0.05), gender (p = 0.34), race (p = 0.31), weight (p = 0.93), height (p = 0.27), and body mass index (p = 0.75); however, the SS group exhibited higher disease duration (96.0 +/- 54.9 vs. 55.2 +/- 52.0 months, p = 0.01). By definition, all PAPS with SS patients suffer from stroke, an arterial event; the frequency of stroke events (28.5 vs. 7.5%, p = 0.04), as well as of limb ischemia (100 vs. 30.0%, p < 0.0001) was higher in this group than in the PAPS without SS group. On the other hand, patients in the PAPS without SS group had more venous events, such as deep venous thrombosis, than those in the PAPS with SS group (80.0 vs. 50.0%, p = 0.03). In conclusion, an understanding of the relationship between APS and SS is important in order to identify a subgroup for which more rigorous accompaniment and therapy may be necessary. PMID- 20012622 TI - Rituximab and concomitant leflunomide for the treatment of rheumatoid arthritis. AB - Rituximab has only been approved in combination with methotrexate for the treatment of rheumatoid arthritis. As some patients have intolerance to methotrexate, alternative co-therapies are needed. This method involved retrospective analysis of ten patients treated with a combination of rituximab and leflunomide. Primary outcome measures were the DAS28 response at month 6 and the time to relapse. The median initial DAS 28 of 5.7 (3.2-7.2) was reduced to 3.5 (1.9-6.1) at month 6. 70% of the patients achieved a good or moderate response, whereas 30% showed no response. Two patients had to stop leflunomide due to adverse effects. Two patients had to reduce the leflunomide dose to 10 mg/day. 5/8 patients experienced a relapse after a median of 10 (6-30) months and were successfully re-treated with rituximab. This small case series suggests that leflunomide might offer an alternative DMARD combination option for the treatment of RA with rituximab. PMID- 20012623 TI - Bowel perforations in a patient affected by Churg-Strauss syndrome under high dose steroid treatment: will alternative drugs reduce risk of surgery? AB - Churg-Strauss syndrome is a relapsing-remitting vasculitis that frequently involves digestive system. Ischemic perforation of the large bowel is relatively rare and potentially life threatening. We report a case treated with high dose of steroids for a relapsing of Churg-Strauss vasculopathy that underwent emergency surgery for multiple large-bowel perforations. Massive use of steroids is common for controlling relapse of this disease, but this increases the risk of intestinal perforation. A prompt switching to alternative drugs when intestinal tract is involved should be considered in order to prevent surgery. PMID- 20012624 TI - Autoantibodies against oxidized low-density lipoprotein and lipid profile in patients with chronic periaortitis: case-control study. AB - Chronic periaortitis is thought to result from an autoallergic reaction to oxidized low-density lipoprotein (OxLDL). No data exist on lipid profile and atherosclerotic biomarkers. We investigated circulating levels of OxLDL and of anti-OxLDL (aOxLDL) antibodies in patients with chronic periaortitis using the cross-sectional case-control study on 20 patients with chronic periaortitis. Patients were compared to 20 age- and sex-matched controls. aOxLDL antibodies were measured by ELISA and expressed as mean optical density values at 450 nm from duplicate measurements (OD(450)). aOxLDL antibody titers (median [interquartile range]) did not differ significantly between patients and controls (aOxLDL-IgM: 0.70 [0.24-1.08] vs. 0.54 [0.25-0.73] OD(450); aOxLDL-IgG: 0.59 [0.38-0.75] vs. 0.41[0.33-0.63]OD(450)). Female patients had higher aOxLDL-IgM levels than male patients (1.02 [0.46-1.38] vs. 0.29 [0.22-0.84] OD(450); P = 0.05). aOxLDL-IgM titers were lower in patients with cardiovascular disease (CVD) than in patients without CVD (0.22 [0.16-0.37] vs. 0.92 [0.70-1.30] OD(450); P = 0.003) and correlated positively with HDL-cholesterol (r = 0.47, 95% CI 0.02 0.69; P = 0.03) and inversely with diastolic blood pressure (r = -0.46, 95% CI 0.75 to -0.01; P = 0.03) and OxLDL/apoB ratio (r = -0.41, 95% CI -0.73 to 0.04; P = 0.06). No differences or associations were found between aOxLDL-IgG titers and other variables between or within patients and/or controls. In patients OxLDL levels correlated with smoking pack-years (r = 0.58, 95% CI 0.17-0.81; P = 0.007). Data suggest a differing innate immune response to OxLDL in patients with chronic periaortitis compared to controls. Whether this response is causally related to chronic periaortitis development remains to be clarified. PMID- 20012625 TI - Malignancy-associated multicentric reticulohistiocytosis. AB - Multicentric reticulohistiocytosis (MR) is a rare systemic disorder of unknown etiology. Up to 25% of patients with MR have associated malignancies. We are reporting a case of a patient with MR who initially had an excellent clinical response to oral methotrexate and prednisone. Subsequent attempts of prednisone taper failed because of disease flare-up. Workup for associated malignancy revealed localized squamous cell carcinoma of the lung. The tumor was resected; methotrexate and prednisone were tapered and discontinued with complete remission of MR. A review of literature and the new advances and treatment modalities for MR is provided. PMID- 20012626 TI - Osteomyelitis variolosa. AB - We present a patient with osteomyelitis variolosa in both elbow joints with short middle fingers-sequelae of smallpox. Both elbow joints showed sclerosis of bones around elbow joint, enlargement of olecranon process of ulna, and head of radius with intact joint space. Hands showed shortening of both middle fingers following dactylitis leading to short metacarpals. The purpose of this report is to remind those who seldom see one of the most important and easily recognized complications of smallpox. PMID- 20012627 TI - Sudden paraplegia following epidural lipomatosis and thoracal compression fracture after long-term steroid therapy: a case report. AB - Sudden paraplegia secondary to the posterior spinal epidural compression and vertebral compression fracture as a complication in corticosteroid treatment is extremely rare. The authors presented a case 49-year-old man with chronic relapsing attack of Still's disease. After the identification of pathology, the surgical evacuation of lipid tissue and pedicle-based instrumentation showed therapeutic success. To the authors' knowledge, this is the first case showing both vertebral fracture and paraplegia that required urgent surgery in the follow up Still's disease. PMID- 20012628 TI - Osteonecrosis secondary to antiphospholipid syndrome: a case report, review of the literature, and treatment strategy. AB - Osteonecrosis is commonly present in patients with antiphospholipid syndrome (APS) and systemic lupus erythematosus (SLE). Treatment of this condition remains extremely controversial. We present a treatment strategy of avascular necrosis of the knee in a patient with catastrophic antiphospholipid syndrome with a history of SLE and APS. Aggressive treatment with 12 rounds of plasmapheresis, intravenous immunoglobulin, rituximab, and cyclophosphamide led to the patient's recovery with no recurrence of symptoms during 16 months of follow up. In this report, we further discuss the pathogenesis of osteonecrosis and current understanding of the treatment of this disease. PMID- 20012629 TI - Programmed cell death 1 gene polymorphisms is associated with ankylosing spondylitis in Chinese Han population. AB - Programmed cell death 1 (PD-1) has been reported to have a genetic association in several autoimmune diseases. The aim of this study was to investigate the association of PD-1 polymorphisms and haplotypes with ankylosing spondylitis (AS) in Chinese Han population. In a case-control association study, three single nucleotide polymorphisms (SNP), PD-1.3 G/A, PD-1.5 C/T and PD-1.9 T/C, were genotyped in 216 AS patients and 264 healthy controls using polymerase chain reaction-restriction fragment length polymorphism assay. All genotype distributions in the patients and in the controls were in Hardy-Weinberg equilibrium. The associations of genotypes and alleles with AS were analyzed. The genotype distributions of PD-1.9 were significantly different between the patients with AS and the controls (P = 0.025). The frequencies of TC genotype and T allele of PD-1.9 were higher in the patients than those in the controls (P = 0.026 and 0.004). No association for PD-1.5 in AS was found, and PD-1.3 was non polymorphic in Chinese Han population. Moreover, the frequency of the CT haplotype (PD-1.5 C/T, PD-1.9 T/C) was significantly higher in AS patients than the controls (21.6 vs. 13.9%, P = 0.002). The CC haplotype was more common in the controls than in the patients (57.1 vs. 44.6%, P = 0.000). The results support a genetic association between the PD-1 polymorphism and susceptibility to AS in Chinese Han population. PMID- 20012630 TI - Regional migratory osteoporosis: case report of a patient with neuropathic pain. AB - Regional migratory osteoporosis (RMO) is an idiopathic disorder characterized by severe periarticular pain, transient and migratory arthralgia, and osteoporosis. Osteoporosis in this disease may appear in the form of local regional osteoporosis and bone marrow edema or generalized osteoporosis. It occurs most commonly in middle-aged men and late second or third trimester pregnant women. The laboratory findings of the disease are usually normal and do not demonstrate apparent anomalies. The presence of bone marrow edema on MRI is its characteristic finding. RMO can only be separated from transient osteoporosis of hip and avascular necrosis with migration to other joints. Clinically, RMO progresses in three stages: increasing pain and disability, radiological findings (osteopenia), maximalization of symptoms, and finally, the regression of the disease and radiological changes. In this case report, we present a 29-year-old woman whose symptoms had first appeared at the second trimester of pregnancy and migrated both to the other joints in the proximo-distal direction and to the adjacent bones within the same joint. She also had symptoms such as hyperalgesia, hyperesthesia and hypertrichosis along with neuropathic pain, which she described as a burning, biting, and prickling type of pain at the right leg. The neuropathic pain of the patient was resistant to medical treatment. We believe that this case was worth reporting because of the obstinate clinical course of the patient's disease and her severe neuropathic pain that was resistant to treatment. PMID- 20012631 TI - Do infections trigger juvenile idiopathic arthritis? AB - Juvenile idiopathic arthritis (JIA) is a disease that was prominent with increased inflammation response in immune system, appeared mostly with peripheral arthritis and endogenous and exogenous antigens play a role in the pathogenesis of disease. Two major reasons were thinking to be considerably important. First of them is immunological predisposition and the second one is environmental factors. Infections are considered to be the most important between environmental factors but also stress and trauma are also important in the etiology of the disease. However, the relation between JIA and infections is not clearly defined but the relation between adult chronic arthritis and infections was well-defined. A total of 70 patients, 26 with primer JIA, 20 with recurrent JIA, 24 healthy control were included in this study. Mycoplasma pneumoniae, Chlamydophila pneumoniae and C. Jejuni were detected in 4, 1 and 1 of 10 (38.46%) patients with primer JIA, respectively. Salmonella enteritidis, EBV, M. pneumoniae, C. jejuni and Borrelia burgdorferi were detected in 1, 2, 2, 2, and 1 of the 8(40%) patients with recurrent JIA, respectively. S. enteritidis were isolated in feces culture and also identified by agglutination method. Infection was detected in total 18 (39.13%) of patient groups. C. pneumoniae and C. jejuni were detected in 1 and 1 of 2(8.33) healthy control groups, respectively. Throat culture positivity was not detected in any of the patient and healthy control groups. In conclusion, etiopathogenesis of JIA is not clearly understood and suggested that various factors can trigger the disease and it is the most common rheumatoid disease of childhood. However, there are some studies focusing especially on one infectious agent but this is the first study including such a big range of infectious agents in the literature for the microorganisms that can be suggested to have a role in the etiopathogenesis of JIA. We have a conclusion in the light of our results and suggest that some microorganisms can trigger and increase the intensity of clinical situation according to the case. When we evaluate the primer and recurrent JIA groups; M. pneumoniae and C. jejuni come forward and seen common in JIA cases. We also suggest that the pre-diagnosis of microorganisms, which can play a role as primarily or by intervening in the etiopathogenesis of JIA and adding specific antimicrobial therapy to the standard JIA therapy, it is possible to perform new, extended, especially molecular based serial case studies. PMID- 20012632 TI - An animal model of chronic rheumatic valvulitis induced by formalin-killed streptococci. AB - Rheumatic heart disease is the most severe complication of rheumatic fever. Till date, very few successful animal models of rheumatic valvular disease have been reported. This study aimed at developing a suitable animal model of chronic rheumatic valvulitis for further investigation and prevention of rheumatic heart disease. Lewis rats were immunized with one administration of formalin-killed and sonicated group A streptococci together with Complete Freund's Adjuvant every 7 days for three cycles followed by group A streptococci alone till killing. Control rats were administered adjuvants and saline. Rats in group 1 were killed 12 weeks after the initial injection. Rats in group 2 and control group were killed 24 weeks after the initial injection. Results 62.5% (5/8) of rats in group 1 developed myocarditis and 50% (4/8) developed valvulitis. Histological examination of cardiac sections showed only cellular infiltrates. In contrast, 75% (6/8) of rats in group 2 developed rheumatic-like myocarditis and 62.5% (5/8) developed chronic valvulitis. Histological manifestations of the hearts in group 2 animals involved not only acute damage such as cellular infiltrates, Aschoff like cells, verrucous vegetation, but also chronic lesions such as fibrosis, vascular neogenesis. None of the rats (0/8) in control group presented myocarditis or valvulitis. Lewis rat repeatedly immunized with formalin-killed GAS may be a suitable animal model of chronic rheumatic valvulitis. It may be useful for future investigation of the pathogenesis and possible preventive strategies of human rheumatic heart disease. PMID- 20012633 TI - The enigmatic roles of microglial versus neuronal progranulin in neurological disease. PMID- 20012634 TI - Biological osteosynthesis of complex proximal humerus fractures: surgical technique and results from a prospective single center trial. AB - INTRODUCTION: Locked plating for complex proximal humerus fractures through a deltopectoral incision can be difficult due to the fracture morphology and need for fixed angle screws. Although good results have been reported with the deltopectoral approach; technical difficulties, excessive soft tissue stripping and fears of avascular necrosis have lead to the use of minimal access techniques. METHOD: Fifteen patients with three or four part fractures were treated by locked plating through a mini-invasive lateral trans-deltoid approach. All patients were relatively young with good bone quality and had sustained a high-velocity injury. Initial closed reduction was attempted in all patients, but majority of the patients (9/15) required open reduction to achieve a satisfactory reduction. RESULTS: Union was achieved in all patients. All fractures united with an acceptable alignment. There were no incidences of axillary nerve palsy. There were no incidences of hardware failure or loss of reduction. There were no incidences of avascular necrosis at 1-year follow-up. The mean normalized constant score at last follow-up was 85.24. CONCLUSION: Locked plating through lateral trans-deltoid incision may offer a better alternative to the deltopectoral approach in these complex fractures where locked plating is contemplated. It respects the fracture biology, allows ease in placement of the locking plate and angle stable screws and offers a stable construct with less surgical morbidity. PMID- 20012635 TI - Prognostic relevance of granulocyte colony-stimulating factor in ovarian carcinomas. AB - PURPOSE: Granulocyte colony-stimulating factor (G-CSF) producing tumours were found associated with poor prognosis. Unfortunately, this finding is based on several case reports only. Thus, we investigated the expression of G-CSF in the tumour cells and the tumour stroma in a large collective of patients with ovarian cancer with long-term follow-up. METHODS: Tissue and clinical records of 175 patients with histologically confirmed ovarian carcinoma were analysed for G-CSF expression in tumour cells and the surrounding stroma. The results were compared with peripheral blood counts and other prognostic factors in ovarian cancer. RESULTS: No correlations were found between both G-CSF expression in tumour cells and the surrounding stroma and prognosis as well as peripheral blood counts. We only found a positive influence of granulocytes in the tumour stroma on prognosis, which however, was not significant in multifactorial analyses. CONCLUSIONS: In contrast to the many case reports from other entities, G-CSF expression in tumour cells and the surrounding stroma is not an adverse prognostic factor. To find out the safety of G-CSF administration for the prevention or treatment of febrile neutropenia, it is suggested for clinical trials to include long-term follow-up and immunohistochemical characterisation of the tumour. PMID- 20012636 TI - Role of HPV testing in the follow-up of women treated for cervical dysplasia. AB - PURPOSE: The aim of study was to investigate factors predicting persistence or relapse of disease after cervical conisation for high-grade squamous intraepithelial lesions (CIN 2 or 3). METHODS: The study involved 78 women with high-grade squamous intraepithelial lesions, conservatively treated with loop electroexcision procedure for cervical conisation and subsequent with CO(2) laser vaporisation of the cervical bed. Histological specimens were totally included and examined by an experienced pathologist. To evaluate the efficacy of treatment, the patients were examined with colposcopy and Pap smear 4 months after surgery and with PCR to search for and genotyping of HPV, 10 months after treatment. RESULTS: During the post-treatment follow-up, the cytologic examination showed persistent/relapsing disease in six patients (7.6%). In only 1 case, the deep margin of the cone was considered positive for CIN (16%).Ten months after treatment, viral typing revealed the persistence of high-risk HPV in all of these patients. Conversely, the viral follow-up of the other 72 patients without persisting/relapsing disease after treatment disclosed low-risk HPV genotypes in 6 cases, high-risk HPV in 2 cases (2.7%), whereas 7 cases had positive margins for CIN (9.7%). The risk of persistence and relapse of CIN in the group with positive margins was not statistically significant (P = 0.87), whereas it was in the group with HR-HPV positive (P = 0.000048). CONCLUSION: HPV testing is the most sensitive mean of identifying persistence or relapse early and is therefore capable of optimising follow-up after the treatment of high grade CIN. PMID- 20012637 TI - The use of Lactobacillus rhamnosus in the therapy of bacterial vaginosis. Evaluation of clinical efficacy in a population of 40 women treated for 24 months. AB - BACKGROUND: Bacterial vaginosis (BV) is the most prevalent vaginal disorder in adult women worldwide. A number of clinical studies indicate that re establishment of the physiological ecosystem by re-colonization of vaginal mucosa by lactobacilli may be an effective therapy for BV both after initial antimicrobial therapy or when given alone. A vaginal pH <4.5 is considered physiological and its measurement has been reported as an objective parameter, very sensitive to alterations of vaginal microflora and correlated with clinically relevant BV. OBJECTIVE: The aims of this work were to assess the effectiveness of a long-term (24 months) intravaginal treatment with Lactobacillus rhamnosus (NORMOGIN) on the vaginal pH and on the clinical symptoms in a group of 40 women affected by BV diagnosed by the Amsel criteria. METHOD: A prospective open clinical trial was performed in 40 consecutive cases evaluated for B.V. by the Amsel criteria. RESULTS: Vaginal pH was above the physiological value of 4.5 in 36 out of 40 patients at the first visit. It returned under 4.5 value in 24/40 and 32/40 women after 12 and 24 months of treatment, respectively. pH values were significantly decreased at 12 month treatment (P < 0.001) and further reduction in pH values was found at 24 months of treatment (P < 0.02 vs. 12 months). The gradual return to a vaginal physiological pH was associated with a reduction of the intensity of symptoms as shown by the decrease in the symptoms score. CONCLUSIONS: The present study supports the use of pH measurement for sensitive, objective, and simple therapy follow-up in women with BV and shows that long-term administration of vaginal tablets containing Lactobacillus rhamnosus represents an effective and safe treatment for restoring the physiological vaginal pH and controlling BV symptoms. PMID- 20012638 TI - Effects of bambuterol and terbutaline on isolated rat's tracheal smooth muscle. AB - Terbutaline (Bricanyl) and its prodrug Bambuterol (Bambec) are highly potent beta(2)-adrenoceptor agonists often used in asthma patients. Terbutaline in the form of inhaled short-acting bronchodilator relieves asthmatic symptoms. However, the effects of bambuterol given intratracheally have rarely been explored. To verify the effect of bambuterol and terbutaline, which act on the tracheal smooth muscle directly in vitro, we used our preparation to test the effects of bambuterol on isolated rat's tracheal smooth muscle compared with terbutaline. The following assessments of bambuterol and terbutaline were performed: (1) effect on tracheal smooth muscle resting tension; (2) effect on contraction caused by 10(-6) M methacholine as a parasympathetic mimetic; (3) effect of the drugs on electrically induced tracheal smooth muscle contractions. The results indicated that adding bambuterol induced a significant further contraction to 10( 6) M methacholine-induced contraction when the preparation was increased to 10( 4) M. But terbutaline elicited a relaxation response at a dose of 10(-6) M or more. Both bambuterol and terbutaline could inhibit electrical field stimulation (EFS) induced spike contraction. Terbutaline had an anti-cholinergic effect that could relieve asthmatic symptoms. But the effect of bambuterol acting on tracheal smooth muscle directly was controversial. PMID- 20012639 TI - Sildenafil therapy and exercise tolerance in idiopathic pulmonary fibrosis. AB - Idiopathic pulmonary fibrosis (IPF) is a form of idiopathic interstitial pneumonia characterized by temporally and spatially heterogeneous fibroblast proliferation and poor prognosis. No therapies have been shown in randomized clinical trials (RCT) to influence survival. Twenty-nine subjects were assigned randomly in a pilot study to a double-blind, placebo-controlled, RCT to test sildenafil in patients with IPF with forced vital capacity 40-90% and diffusing capacity 30-90% of predicted. During the 6-month experimental treatment period, patients underwent 6-min walk tests and estimation of dyspnea using the Borg scale at baseline (0 months), 3 months, and 6 months. Participants had moderate impairment of pulmonary function, and there were no significant differences between placebo (n = 15) and sildenafil (n = 14)-treated groups. Sildenafil did not significantly increase 6-min walk test distance (mean distance +/- SD after 6 month protocol: placebo 355 +/- 82 m, sildenafil 324 +/- 41 m; p = 0.256) nor did it lessen dyspnea after exercise (mean Borg score after 6-month protocol: placebo 3.4 +/- 1.6, sildenafil 4.1 +/- 2.3; p = 0.492). Adverse reactions were few and minor in nature. In this trial, sildenafil did not significantly increase 6-min walk test distance or decrease the Borg dyspnea index in patients with clinically typical IPF. This trial was registered at clinicaltrials.gov as NCT00359736. PMID- 20012640 TI - Cheyne-stokes respiration and prognosis in modern-treated congestive heart failure. AB - In patients with congestive heart failure (CHF), a high prevalence of sleep disordered breathing has been described. Cheyne-Stokes respiration (CSR) is present in up to 40% of patients with CHF. During the last decade, the medical treatment has been substantially improved. This study was designed to analyze the prognosis of CSR in modern-treated patients with CHF. For this purposes, in 57 patients with CHF who received modern treatment, a 5-year follow-up after initial full night polysomnography was performed. The mean follow-up period was 38 +/- 18 months. Mean age was 62 +/- 13 years and the mean ejection fraction was 25 +/- 7 percent. Respiratory polygraphy revealed CSR with a respiratory disturbance index >5 per hour of sleep in 39 of 57 patients. Twelve patients died. CSR was only characterized by a tendency of worsening (log-rank test, p = 0.25). However, there was a significant difference toward positive outcome for patients who received cardiac resynchronization therapy (log-rank test, p = 0.036). Using Multivariate Cox's proportional hazard regression with the factors resynchronization and CSR, the effect of resynchronization was almost significant (p = 0.08). In conclusion, no significant change of Cheyne-Stokes prevalence can be found in our small group of modern-treated patients with CHF. Cardiac resynchronization therapy was associated with improved patient outcome. PMID- 20012641 TI - Impulse oscillometry and respiratory symptoms in World Trade Center responders, 6 years post-9/11. AB - This study evaluated whether impulse oscillometry (IOS) testing revealed signs of respiratory disease in New York State (NYS) World Trade Center (WTC) responders in comparison with unexposed NYS employees. It also compared self-reported respiratory symptoms between the two groups, 6 years post-9/11. For this evaluation participants completed a self-administered questionnaire regarding respiratory symptoms. IOS testing included measures of resistance and reactance to assess for peripheral versus central airway effects. Two hundred forty-eight subjects (99 exposed and 149 unexposed) were included in the final analysis. Since September 11, 2001, NYS responders were more likely to report new or worsening cough in the absence of a respiratory infection, cough consistent with chronic bronchitis, current respiratory symptoms, or lower respiratory symptoms in the last 12 months. Significant associations were found between IOS indices and gender, smoking history, and obesity. When comparing exposed and unexposed participants, there were no significant differences in the geometric means of the IOS indices. Responders who used a respirator with canister demonstrated significantly lower respiratory resistance at 5 and 20 Hz (R5 and R20). While this study has provided no evidence of an association between WTC exposure and peripheral airways disease in this cohort of responders, results do suggest that use of a respirator with canister may be protective for central airways in responders exposed to dust and smoke. This emphasizes the importance of stressing proper respirator use in planning responses to future disasters. Our control data also provide useful reference values for future IOS research. PMID- 20012642 TI - Bone spicule pigment formation in retinitis pigmentosa: insights from a mouse model. AB - BACKGROUND: Bone spicule pigments (BSP) are a hallmark of retinitis pigmentosa (RP). In this study, we examined the process of BSP formation in the rhodopsin knockout (rho (-/-)) mouse, a murine model for human RP. METHODS: In rho (-/-) mice from 2 to 16 months of age, representing the range from early to late stages of degeneration, retinal sections and whole mounts were examined morphologically by light and electron microscopy. The results were compared to scanning laser ophthalmoscopy of BSP degeneration in human RP. RESULTS: After the loss of all photoreceptor cells in rho-/- mice, the outer retina successively degenerated, leading to approximation and finally a direct contact of inner retinal vessels and the retinal pigment epithelium (RPE). We could show that it was the event of proximity of retinal vessel and RPE that triggered migration of RPE cells along the contacting vessels towards the inner retina. Ultrastructurally, these mislocalized RPE cells partially sealed the vessels by tight junction linkage and deposited extracellular matrix perivascularly. Also, the vascular endothelium developed fenestrations similar to the RPE-choroid interface. In whole mounts, the pigmented cell clusters outlining retinal capillaries correlated well with BSPs in human RP. The structure of the inner retina remained well preserved, even in late stages. CONCLUSIONS: The Rho (-/-) mouse is the first animal model that depicts all major pathological changes, even in the late stages of RP. Using the rho (-/-) mouse model we were able to analyze the complete dynamic process of BSP formation. Therefore we conclude that: (1) In rho (-/-) retinas, BSPs only form in areas devoid of photoreceptors; (2) Direct contact between inner retinal vessels and RPE appears to be a major trigger for migration of RPE cells; (3) The distribution of the RPE cells in BSPs reflects the vascular network at the time of formation. The similarity of the disease process between mouse and human and the possibility to study all consecutive steps of the course of the disease makes the rho (-/-) mouse valuable for further insights in the dynamics of BSP formation in human RP. PMID- 20012643 TI - Intravitreal injection of bevacizumab and triamcinolone acetonide at the end of vitrectomy for diabetic vitreous hemorrhage: a comparative study. AB - BACKGROUND: To study the clinical outcome and complications of intravitreal bevacizumab (IVB) versus triamcinolone acetonide (IVT) injections at the end of vitrectomy in patients with diabetic vitreous hemorrhage. METHODS: This is a retrospective comparative case study comprised of 156 eyes from 139 consecutive patients who underwent pars plana vitrectomy for diabetic vitreous hemorrhage (VH) with or without tractional retinal detachment (TRD). IVB (1.25 mg/0.05 ml) injection was done for 66 eyes (60 patients) and IVT (4 mg/0.1 ml) injection for 33 eyes (31 patients) at the end of vitrectomy, and no injection was done for 57 eyes (48 patients). Main outcome measures were best-corrected visual acuity (BCVA), intraocular pressure (IOP), and incidence of postoperative VH and reoperation with at least 12 months of follow-up. RESULTS: Early postoperative VH within 1 month postoperatively occurred in eight eyes (12.1%) from the IVB group, three eyes (9.1%) from the IVT group, and 21 eyes (36.8%) from the control group. The rate of early postoperative VH was significantly lower in the IVB and IVT group than the control group (p = 0.002 and 0.006, respectively). Late postoperative VH after 1 month postoperatively occurred in 11 eyes (16.7%) from the IVB group, three eyes (9.1%) from the IVT group, and 12 eyes (21.1%) from the control group. There was no difference between the three groups (P = 0.341). BCVA of the IVB, IVT, and control group at postoperative months 1, 3, 6, 9, and 12 significantly improved from the preoperative BCVA (p < 0.01, respectively). There was no difference for BCVA between the three groups at each follow-up period (p > 0.05). In only the IVT group, IOP at postoperative day 1 was higher than IOP preoperatively (p = 0.002). There was no significant difference in the rate of reoperation between the three groups (p = 0.275). CONCLUSION: IVB and IVT injection at the end of diabetic vitrectomy is a useful adjunctive therapy for reducing early postoperative VH compared to the control group. IVB injection does not have a risk of a temporary rise of IOP, which the IVT group has. PMID- 20012644 TI - Presumed endophthalmitis following Boston keratoprosthesis treated with 25 gauge vitrectomy: a report of three cases. AB - BACKGROUND: Although endophthalmitis is known to occur frequently in patients with KPros, the management remains a challenge both due to the lack of evidence and the exceptionality of the examination and the therapeutic approach. MATERIALS AND METHODS: Three patients implanted with Boston keratoprosthesis for management of ocular cicatricial pemphigoid and severe alkali injury presented with severe late-onset (10-24 months post-operatively), presumed bacterial endophthalmitis. Three-port 25-gauge (g) pars plana vitrectomy was performed in all patients; vitreous samples were acquired and intravitreal vancomycin and cefuroxime were administered. RESULTS: Post-operative outcome was significantly improved in all three cases. Visual acuity improved from perception of light (PL) to 20/400 in the first, PL to counting fingers in the second and PL to 20/400 in the third patient. CONCLUSION: We present our experience from managing presumed bacterial endophthalmitis with 25 g PPV in patients previously implanted with Boston Kpro, which to the best of our knowledge has not been reported before. Although the number of cases is small to draw accurate conclusions, we have to report that 25 g vitrectomy was performed without significant difficulties and seems to be a useful tool for the management of patients with Kpros requiring vitreoretinal surgery. PMID- 20012645 TI - Age-dependent axonal expression of potassium channel proteins during development in mouse hippocampus. AB - The development of the hippocampal network requires neuronal activity, which is shaped by the differential expression and sorting of a variety of potassium channels. Parallel to their maturation, hippocampal neurons undergo a distinct development of their ion channel profile. The age-dependent dimension of ion channel occurrence is of utmost importance as it is interdependently linked to network formation. However, data regarding the exact temporal expression of potassium channels during postnatal hippocampal development are scarce. We therefore studied the expression of several voltage-gated potassium channel proteins during hippocampal development in vivo and in primary cultures, focusing on channels that were sorted to the axonal compartment. The Kv1.1, Kv1.2, Kv1.4, and Kv3.4 proteins showed a considerable temporal variation of axonal localization among neuronal subpopulations. It is possible, therefore, that hippocampal neurons possess cell type-specific mechanisms for channel compartmentalization. Thus, age-dependent axonal sorting of the potassium channel proteins offers a new approach to functionally distinguish classes of hippocampal neurons and may extend our understanding of hippocampal circuitry and memory processing. PMID- 20012648 TI - Neutrophils promote inflammatory angiogenesis via release of preformed VEGF in an in vivo corneal model. AB - We investigated the role of neutrophilic cells (neutrophils) in inflammatory angiogenesis and explored the possible mechanisms involved. Corneal angiogenesis was induced in vivo with a 75% silver nitrate applicator. Depletion of neutrophils was accomplished by the intraperitoneal administration of RB6-8C5, a neutrophil-depleting antibody. Angiogenesis, neutrophil infiltration, and the localization of vascular endothelial growth factor (VEGF) were evaluated by biomicroscopic observations, histology, and immunohistochemistry in control and RB6-8C5 treatment groups. Protein levels of VEGF, macrophage inflammatory protein 1alpha (MIP-1alpha), macrophage inflammatory protein-2 (MIP-2), and tumor necrosis factor alpha in the cornea were determined by enzyme-linked immunosorbent assay. An in vitro model of neutrophil activation was also used to examine the ability of neutrophils to produce and release VEGF, MIP-1alpha, and MIP-2. At day 1 after injury, neutrophil infiltration in the cornea was highest, and VEGF was expressed in the infiltrating neutrophils. The enhanced protein levels of VEGF, MIP-1alpha, and MIP-2 correlated with the degree of neutrophil infiltration. Neutrophil depletion significantly inhibited corneal angiogenesis and reduced the protein levels of VEGF, MIP-1alpha, and MIP-2 in the cornea. Upon stimulation, isolated neutrophils released VEGF from preformed stores and MIP 1alpha and MIP-2 by de novo synthesis. Neutrophil depletion thus significantly impaired inflammatory angiogenesis, identifying neutrophils as an important player in inflammatory angiogenesis. Neutrophils may exercise their angiogenic function by releasing proangiogenic factors such as VEGF. Intervention measures targeting neutrophils may therefore help to deal with abnormal angiogenesis involved in chronic inflammatory diseases. PMID- 20012647 TI - Clinical practice: Diagnosis and management of Henoch-Schonlein purpura. AB - Henoch-Schonlein purpura (HSP) is the most common vasculitis of childhood. In this review, the main clinical features and complications are described. Although most features are self-limiting, renal disease is the most likely to result in long-term morbidity. Treatment of HSP nephritis is controversial, and the evidence for both prevention and treatment of established disease is reviewed. Follow-up for children presenting with HSP should be for at least 6 months and should include regular urine testing for proteinuria and haematuria and a blood pressure measurement. Women with a history of HSP during childhood are at increased risk of complications (such as proteinuria and hypertension) during pregnancy and should be monitored closely. This paper describes current practice with regard to investigation and management and proposes a practical care pathway of the management of a child with HSP. PMID- 20012649 TI - Effects of introducing cultured human chondrocytes into a human articular cartilage explant model. AB - Articular cartilage (AC) heals poorly and effective host-tissue integration after reconstruction is a concern. We have investigated the ability of implanted chondrocytes to attach at the site of injury and to be incorporated into the decellularized host matrix adjacent to a defect in an in vitro human explant model. Human osteochondral dowels received a standardized injury, were seeded with passage 3 chondrocytes labelled with PKH 26 and compared with two control groups. All dowels were cultured in vitro, harvested at 0, 7, 14 and 28 days and assessed for chondrocyte adherence and migration into the region of decellularized tissue adjacent to the defects. Additional evaluation included cell viability, general morphology and collagen II production. Seeded chondrocytes adhered to the standardized defect and areas of lamina splendens disruption but did not migrate into the adjacent acellular region. A difference was noted in viable-cell density between the experimental group and one control group. A thin lattice-like network of matrix surrounded the seeded chondrocytes and collagen II was present. The results indicate that cultured human chondrocytes do indeed adhere to regions of AC matrix injury but do not migrate into the host tissue, despite the presence of viable cells. This human explant model is thus an effective tool for studying the interaction of implanted cells and host tissue. PMID- 20012650 TI - Proceedings of the 8th Symposium on Growth and Nutrition in Children with Chronic Renal Disease, Symposium of the International Pediatric Nephrology Association (IPNA). Oviedo, Spain. May, 28-30, 2009. PMID- 20012651 TI - Therapeutic intervention and surgery of acute pancreatitis. AB - The clinical course of acute pancreatitis varies from mild to severe. Assessment of severity and etiology of acute pancreatitis is important to determine the strategy of management for acute pancreatitis. Acute pancreatitis is classified according to its morphology into edematous pancreatitis and necrotizing pancreatitis. Edematous pancreatitis accounts for 80-90% of acute pancreatitis and remission can be achieved in most of the patients without receiving any special treatment. Necrotizing pancreatitis occupies 10-20% of acute pancreatitis and the mortality rate is reported to be 14-25%. The mortality rate is particularly high (34-40%) for infected pancreatic necrosis that is accompanied by bacterial infection in the necrotic tissue of the pancreas (Widdison and Karanjia in Br J Surg 80:148-154, 1993; Ogawa et al. in Research of the actual situations of acute pancreatitis. Research Group for Specific Retractable Diseases, Specific Disease Measure Research Work Sponsored by Ministry of Health, Labour, and Welfare. Heisei 12 Research Report, pp 17-33, 2001). On the other hand, the mortality rate is reported to be 0-11% for sterile pancreatic necrosis which is not accompanied by bacterial infection (Ogawa et al. 2001; Bradely and Allen in Am J Surg 161:19-24, 1991; Rattner et al. in Am J Surg 163:105-109, 1992). The Japanese (JPN) Guidelines were designed to provide recommendations regarding the management of acute pancreatitis in patients having a variety of clinical characteristics. This article describes the guidelines for the surgical management and interventional therapy of acute pancreatitis by incorporating the latest evidence for the management of acute pancreatitis in the Japanese-language version of JPN guidelines 2010. Eleven clinical questions (CQ) are proposed: (1) worsening clinical manifestations and hematological data, positive blood bacteria culture test, positive blood endotoxin test, and the presence of gas bubbles in and around the pancreas on CT scan are indirect findings of infected pancreatic necrosis; (2) bacteriological examination by fine needle aspiration is useful for making a definitive diagnosis of infected pancreatic necrosis; (3) conservative treatment should be performed in sterile pancreatic necrosis; (4) infected pancreatic necrosis is an indication for interventional therapy. However, conservative treatment by antibiotic administration is also available in patients who are in stable general condition; (5) early surgery for necrotizing pancreatitis is not recommended, and it should be delayed as long as possible; (6) necrosectomy is recommended as a surgical procedure for infected necrosis; (7) after necrosectomy, a long-term follow-up paying attention to pancreatic function and complications including the stricture of the bile duct and the pancreatic duct is necessary; (8) drainage including percutaneous, endoscopic and surgical procedure should be performed for pancreatic abscess; (9) if the clinical findings of pancreatic abscess are not improved by percutaneous or endoscopic drainage, surgical drainage should be performed; (10) interventional treatment should be performed for pancreatic pseudocysts that give rise to symptoms, accompany complications or increase the diameter of cysts and (11) percutaneous drainage, endoscopic drainage or surgical procedures are selected in accordance with the conditions of individual cases. PMID- 20012646 TI - Vibration as an exercise modality: how it may work, and what its potential might be. AB - Whilst exposure to vibration is traditionally regarded as perilous, recent research has focussed on potential benefits. Here, the physical principles of forced oscillations are discussed in relation to vibration as an exercise modality. Acute physiological responses to isolated tendon and muscle vibration and to whole body vibration exercise are reviewed, as well as the training effects upon the musculature, bone mineral density and posture. Possible applications in sports and medicine are discussed. Evidence suggests that acute vibration exercise seems to elicit a specific warm-up effect, and that vibration training seems to improve muscle power, although the potential benefits over traditional forms of resistive exercise are still unclear. Vibration training also seems to improve balance in sub-populations prone to fall, such as frail elderly people. Moreover, literature suggests that vibration is beneficial to reduce chronic lower back pain and other types of pain. Other future indications are perceivable. PMID- 20012652 TI - Fundamental and intensive care of acute pancreatitis. AB - Patients who have been diagnosed as having acute pancreatitis should be, on principle, hospitalized. Crucial fundamental management is required soon after a diagnosis of acute pancreatitis has been made and includes monitoring of the conscious state, the respiratory and cardiovascular system, the urinary output, adequate fluid replacement and pain control. Along with such management, etiologic diagnosis and severity assessment should be conducted. Patients with a diagnosis of severe acute pancreatitis should be transferred to a medical facility where intensive respiratory and cardiovascular management as well as interventional treatment, blood purification therapy and nutritional support are available. The disease condition in acute pancreatitis changes every moment and even symptoms that are mild at the time of diagnosis may become severe later. Therefore, severity assessment should be conducted repeatedly at least within 48 h following diagnosis. An adequate dose of fluid replacement is essential to stabilize cardiovascular dynamics and the dose should be adjusted while assessing circulatory dynamics constantly. A large dose of fluid replacement is usually required in patients with severe acute pancreatitis. Prophylactic antibiotic administration is recommended to prevent infectious complications in patients with severe acute pancreatitis. Although the efficacy of intravenous administration of protease inhibitors is still a matter of controversy, there is a consensus in Japan that a large dose of a synthetic protease inhibitor should be given to patients with severe acute pancreatitis in order to prevent organ failure and other complications. Enteral feeding is superior to parenteral nutrition when it comes to the nutritional support of patients with severe acute pancreatitis. The JPN Guidelines recommend, as optional continuous regional arterial infusion and blood purification therapy. PMID- 20012653 TI - Helicobacter pylori status and endoscopic findings in Japan. PMID- 20012654 TI - Comparison of HCV-associated gene expression and cell signaling pathways in cells with or without HCV replicon and in replicon-cured cells. AB - BACKGROUND: Hepatitis C virus (HCV) replication is affected by several host factors. Here, we screened host genes and molecular pathways that are involved in HCV replication by comprehensive analyses using two genotypes of HCV replicon expressing cells, their cured cells and naive Huh7 cells. METHODS: Huh7 cell lines that stably expressed HCV genotype 1b or 2a replicon were used. The cured cells were established by treating HCV replicon cells with interferon-alpha. Expression of 54,675 cellular genes was analyzed by GeneChip DNA microarray. The data were analyzed by using the KEGG Pathway database. RESULTS: Hierarchical clustering analysis showed that the gene-expression profiles of each cell group constituted clear clusters of naive, HCV replicon-expressed, and cured cell lines. The pathway process analysis between the replicon-expressing and the cured cell lines identified significantly altered pathways, including MAPK, steroid biosynthesis and TGF-beta signaling pathways, suggesting that these pathways were affected directly by HCV replication. Comparison of cured and naive Huh7 cells identified pathways, including steroid biosynthesis and sphingolipid metabolism, suggesting that these pathways were required for efficient HCV replication. Cytoplasmic lipid droplets were obviously increased in replicon-expressing and cured cells as compared to naive cells. HCV replication was significantly suppressed by peroxisome proliferator-activated receptor (PPAR)-alpha agonists but augmented by PPAR-gamma agonists. CONCLUSION: Comprehensive gene expression and pathway analyses show that lipid biosynthesis pathways are crucial to support proficient virus replication. These metabolic pathways could constitute novel antiviral targets against HCV. PMID- 20012655 TI - Ectomycorrhizal fungi associated with two species of Kobresia in an alpine meadow in the eastern Himalaya. AB - The diversity of ectomycorrhizal fungi (EMF) on Kobresia filicina and Kobresia capillifolia in an alpine meadow in China's southwestern mountains, one of the word's hotspots of biodiversity, was estimated based on internal transcribed spacer rDNA sequence analysis of root tips. Seventy EMF operational taxonomical units (OTUs) were found in the two plant species. Dauciform roots with EMF were detected in species of Kobresia for the first time. OTU richness of EMF was high in Tomentella/Thelophora and Inocybe, followed by Cortinarius, Sebacina, the Cenococcum geophilum complex, and Russula. Tomentella/Thelophora and Inocybe were general and dominant mycobiont genera of the two sedges. Besides the C. geophilum complex, the ascomycete components Hymenoscyphus and Lachnum were also detected on the two plants. Alpine plants in different geographical regions share similar main genera and/or families of EMF while harboring predominantly different mycobiont species; most of the members detected by us have not been found elsewhere. Significant differences in the profile of EMF occurrences were not found between the two plant species and among the three sampling seasons in our sample size. PMID- 20012656 TI - Erythropoietin concentration in serum and cerebrospinal fluid of patients with amyotrophic lateral sclerosis. AB - Erythropoietin (EPO) acts as a neuroprotective factor and is upregulated after neuronal injury. It has been reported that in cerebrospinal fluid (CSF) of amyotrophic lateral sclerosis (ALS) patients, the EPO concentration is decreased. In this study, EPO levels in serum and CSF of 30 patients with ALS and in 15 controls, using an ELISA technique, were estimated. EPO level in serum was decreased, especially in patients with bulbar onset ALS. A trend toward a progressive EPO decline with the duration of the disease in the mild + moderate ALS cases was observed. In severe cases, a tendency towards a positive correlation of EPO and duration of the disease was present. Serum EPO values were age related only in mild + moderate ALS in patients below 40 years of age. In CSF, the EPO levels were significantly decreased. Lower EPO values in the bulbar onset ALS when compared with the spinal onset ALS were present. The EPO decrease did not correlate with the severity and duration of the disease. Age relation of the EPO level only in the mild + moderate ALS cases more than 40 years was present. Lack of differences in EPO levels between patients with ALS of rapid and slow progression indicates that EPO concentration cannot be used as a prognostic factor. Nevertheless, the decreased serum and CSF EPO concentration and the known EPO neuroprotective action may indicate that EPO administration can be a new promising therapeutic approach in ALS. PMID- 20012658 TI - High stability of original predominant pospiviroid genotypes upon mechanical inoculation from ornamentals to potato and tomato. AB - Eleven pospiviroid isolates from ornamental plants and one from pepper were successfully transmitted to potato and tomato by mechanical inoculation. All isolates with characteristic predominant genotypes were inoculated to a series of potato and tomato plants and propagated for up to four passages. In total, 385 nucleotide sequences were determined, in which 17 new predominant genotypes were identified with minimal differences to the original predominant genotype. In addition, in the original ornamental hosts, only one of eleven predominant pospiviroid genotypes had changed during the experiments over a period of 2 years. These results confirm the high stability of predominant pospiviroid genotypes. PMID- 20012657 TI - Short- and long-term dopaminergic effects on dysarthria in early Parkinson's disease. AB - While the beneficial effect of levodopa on motor impairment in Parkinson's disease (PD) has been well documented, its effect on speech has rarely been examined and the respective literature is inconclusive. The aim of our study was to analyze the effect of short-term levodopa admission and long-term dopaminergic treatment on speech in PD patients in early stages of the disease. Motor examination according to UPDRS III and speech testing were performed in 23 PD patients (9 males; median age 68, 42-78 years) in the early morning after having abstained from dopaminergic medication overnight ("off" state, t0) after administration of 200 mg of soluble levodopa (t1), and at follow-up after 12-14 weeks under stable dopaminergic medication (t2). Speech examination comprised the perceptual rating of global speech performance and an acoustical analysis based upon a standardized reading task. While UPDRS III showed a significant amelioration after L: -dopa application, none of the parameters of phonation, intonation, articulation and speech velocity improved significantly in the "on" state, neither under short-term levodopa administration (t1) nor on stable dopaminergic treatment (t2). However, there was a positive effect of dopaminergic stimulation on vowel articulation in individual patients. Results indicated significant beneficial effect of short-term levodopa administration or long-term dopaminergic medication on different dimensions of speech in PD patients. As some improvement of vowel articulation was seen in individual patients, the pre existing pattern of speech impairment might be responsible for the different response to pharmacological treatment. PMID- 20012659 TI - In memoriam: Jonathan S. Allan, DVM (1952-2009). PMID- 20012661 TI - Energy metabolism of Inuit sled dogs. AB - We explored how seasonal changes in temperature, exercise and food supply affected energy metabolism and heart rate of Inuit dogs in Greenland. Using open flow respirometry, doubly labeled water, and heart rate recording, we measured metabolic rates of the same dogs at two different locations: at one location the dogs were fed with high energy food throughout the year while at the other location they were fed with low energy food during summer. Our key questions were: is resting metabolic rate (RMR) increased during the winter season when dogs are working? Does feeding regime affect RMR during summer? What is the proportion of metabolic rate (MR) devoted to specific dynamic action (SDA), and what is the metabolic scope of working Inuit sled dogs? The Inuit dogs had an extremely wide thermoneutral zone extending down to -25 degrees C. Temperature changes between summer and winter did not affect RMR, thus summer fasting periods were defined as baseline RMR. Relative to this baseline, summer MR was upregulated in the group of dogs receiving low energy food, whereas heart rate was downregulated. However, during food digestion, both MR and HR were twice their respective baseline values. A continuously elevated MR was observed during winter. Because temperature effects were excluded and because there were also no effects of training, we attribute winter elevated MR to SDA because of the continuous food supply. Working MR during winter was 7.9 times the MR of resting dogs in winter, or 12.2 times baseline MR. PMID- 20012660 TI - Molecular and functional characterization of the cystic fibrosis transmembrane conductance regulator from the Australian common brushtail possum, Trichosurus vulpecula. AB - Unlike eutherian mammals, the colon of the Australian common brushtail possum, Trichosurus vulpecula, a metatherian mammal, is incapable of electrogenic Cl(-) secretion and has elevated levels of electrogenic Na(+) absorption, while the ileum secretes HCO (3) (-) rather than Cl(-). In eutherian mammals, the cystic fibrosis transmembrane conductance regulator (CFTR) is essential for both Cl(-) and HCO (3) (-) secretion and the regulation of Na(+) absorption. Therefore, we have sequenced possum (p)CFTR, described its distribution and characterized the properties of cloned pCFTR expressed in Fischer rat thyroid (FRT) cells. pCFTR (GenBank accession No. AY916796) has a 1,478 amino acid open reading frame, which has >90% identity with CFTR from other marsupials and >80% identity with non rodent eutherian mammals. In pCFTR, there is a high level of conservation of the transmembrane and nucleotide binding domains although, with the exception of other marsupials, there is considerable divergence from other species in the R domain. FRT cells transfected with pCFTR express mature CFTR protein which functions as a small Cl(-) channel activated by cAMP-dependent phosphorylation. In whole-cell recordings it has a linear, time and voltage-independent conductance, with a selectivity sequence P(Br) > P(Cl) > P(I) > P(HCO)(3) >> P(Gluconate). pCFTR transcript is present in a range of epithelia, including the ileum and the colon. The presence of pCFTR in the ileum and its measured HCO (3) (-) permeability suggest that it may be involved in ileal HCO (3) (-) secretion. Why the possum colon does not secrete Cl(-) and has elevated electrogenic Na(+) absorption, despite the apparent expression of CFTR, remains to be determined. PMID- 20012662 TI - The effects of progressive hypoxia and re-oxygenation on cardiac function, white muscle perfusion and haemoglobin saturation in anaesthetised snapper (Pagrus auratus). AB - The effects of progressive hypoxia and re-oxygenation on cardiac function, white muscle perfusion and haemoglobin saturation were investigated in anaesthetised snapper (Pagrus auratus). White muscle perfusion and haemoglobin saturation were recorded in real time using fibre optic methodology. A marked fall in heart rate (HR) was evoked when the water bath dissolved oxygen (DO) concentration decreased below 1.5 mg L(-1). This bradycardia deepened over the subsequent 20 min of progressive hypoxia and noticeable arrhythmias occurred, suggesting that hypoxia had direct and severe effects on the cardiac myocytes. Perfusion to the white muscle decreased below a DO concentration of 3 mg L(-1), and oxyhaemoglobin concentration decreased once the DO fell below ca. 2 mg L(-1). During re oxygenation, heart rate and white muscle perfusion increased as the DO concentration exceeded 1.9 +/- 0.1 mg L(-1), whereas haemoglobin saturation increased once the external DO concentration reached 2.9 mg L(-1). These changes occurred in anaesthetised fish, in which sensory function must be impaired, if not abolished. As white muscle perfusion both fell and increased prior to changes in white muscle oxyhaemoglobin saturation, a local hypoxia is more likely to be the consequence than the cause of the reduced blood delivery, and changes upstream from the tail vasculature must be responsible. HR and tissue haemoglobin concentrations did increase simultaneously on re-oxygenation suggesting an increased cardiac output as the cause. PMID- 20012663 TI - Mean platelet volume in psoriasis and psoriatic arthritis. AB - Mean platelet volume (MPV), an indicator of platelet activation, is a newly emerging risk factor for atherothrombosis. There is evidence of platelet activation in psoriasis and psoriatic arthritis (PsA). The association between psoriasis, PsA, and atherosclerosis is well documented, yet, the underlying mechanisms remain unclear. The aim of this study was to investigate the differences of MPV values in patients with psoriasis, PsA, and healthy subjects and the correlation between MPV and the clinical disease activity. A total of 106 patients with psoriasis were included in this study. The study population grouped as 48 patients with PsA (group 1) and 58 patients without PsA (group 2) and 95 healthy controls (group 3). MPV was measured in psoriasis and PsA patients. MPV values were collected from standard complete blood count samples. Clinical features and PASI scores in group 1 and 2 were also recorded. MPV in patients with psoriasis 8.7 +/- 0.9 fL was significantly higher than that of control subjects 7.3 +/- 0.8 fL (p < 0.001). There was also statistical difference between MPV levels of patients with (9.5 +/- 0.8) and without (8.0 +/- 0.7) arthritis (p < 0.001). MPV levels were positively correlated with psoriasis area and severity index score (p = 0.000, r = +0.735). MPV levels showed positive correlation with disease duration (p = 0.01, r = 0.518). MPV levels are increased in patients with psoriasis and PsA. MPV may be a marker for the severity of psoriasis. This study may confirm previous observation indicating increased platelet activation in psoriasis. Increased platelet activity could contribute to increasing the atherosclerotic risk in patients with psoriasis and PsA. PMID- 20012665 TI - Line tension and the shape of nanodroplets. AB - Various factors are discussed which might influence the equilibrium contact angle of nanodroplets placed on a solid substrate. Special emphasis is put on the possible role of the dependence of the solid-liquid interface tension [Formula: see text] on the pressure in the liquid, which in nanodrops considerably exceeds the saturation pressure. We show that certain published data regarding that dependence are meaningless because these have been deduced based on an inconsistent data analysis. PMID- 20012664 TI - Recruitment of closely linked genes for divergent functions: the seed storage protein (Glu-3) and powdery mildew (Pm3) genes in wheat (Triticum aestivum L.). AB - Wheat seed storage protein gene loci (Glu-3) and powdery mildew resistance gene loci (Pm3 and Pm3-like) are closely linked on the short arms of homoeologous group 1 chromosomes. To study the structural organization of the Glu-3/Pm3 loci, three bacterial artificial chromosome clones were sequenced from the A, B, and D genomes of hexaploid wheat. The A and B genome clones contained a Glu-3 adjacent to a Pm3-like gene organized in a conserved Glu-3/SFR159/Pm3-like structure. The D genome clone contained clusters of resistance gene analogs but no Pm3. Its similarity to the A and B genome was limited to the Glu-3/SFR159 region. Comparison of the B genome PM3-like deduced amino acid sequence with known PM3 functional isotypes reinforced the hypothesis of allelic evolution via block exchange by gene conversion/recombination. The advent of glutenin genes and the formation of the Glu-3/SFR159/Pm3 locus occurred after divergence of wheat from rice and Brachypodium. Comparison of the A genome homologous sequences permitted an estimate of time of divergence of approximately 0.3 million years ago. The B genome sequences were not colinear indicating that they could either be paralogs or represent different B genome progenitors. Analysis of the 11 complete retrotransposons indicated a time of divergence ranging from 0.29 to 5.62 million years ago, consistent with their complex nested structure. PMID- 20012666 TI - High-resolution ellipsometric studies on fluid interfaces. AB - In this article, highly accurate experimental results reveal the interfacial profile between different macroscopic fluid phases. The deviation from a step profile, quantified by the ellipsometric quantity J(1), shows a strong correlation with the cohesive energy quantified by the Gordon parameter G . Surprisingly, at high values of G , J (1)( < 0) deviates significantly from any predictions. Findings for water and water-like interfaces can be interpreted in terms of the strength of hydrogen bonding at the surface. PMID- 20012667 TI - Towards a structural characterization of charge-driven polymer micelles. AB - Light scattering and small-angle neutron scattering experiments were performed on comicelles of several combinations of oppositely charged (block co)polymers in aqueous solutions. Fundamental differences between the internal structure of this novel type of micelle --termed complex coacervate core micelle (C3Ms), polyion complex (PIC) micelle, block ionomer complex (BIC), or interpolyelectrolyte complex (IPEC)-- and its traditional counterpart, i.e., a micelle formed via self assembly of polymeric amphiphiles, give rise to differences in scaling behaviour. Indeed, the observed dependencies of micellar size and aggregation number on corona block length, N (corona) , are inconsistent with scaling predictions developed for polymeric micelles in the star-like and crew-cut regime. Generic C3M characteristics, such as the relatively high core solvent fraction, the low core-corona interfacial tension, and the high solubility of the coronal chains, are causing the deviations. A recently proposed scaling theory for the cross-over regime, as well as a primitive first-order self-consistent field (SCF) theory for obligatory co-assembly, follow our data more closely. PMID- 20012668 TI - Colorectal cancer in Turkey: current situation and challenges for the future. AB - Cancer is foremost among one of the major public health problems in Turkey. Projections for the year 2030 estimate that cancer will take the highest death toll for both males and females. In line with the dynamic health care reform process commencing in 2003, the national cancer policy of Turkey has also changed radically. Cancer became more visible as a public health problem and improvements have been made in early diagnosis and treatment of the disease. In addition, major steps have been taken to improve the cancer registry system in order to improve cancer statistics. Colorectal cancer (CRC) incidence is approximately 7 per 100,000, with approximately 5,000 new cases and 3,200 deaths annually. Although cancer treatment expenditures are widely covered in Turkey, there are still problems related to the general accessibility of the health care system. This paper addresses the case of cancer in Turkey with special focus on CRC. Specifically, detailed information is provided about the health care and cancer policies, improvements since 2003 and challenges for the future. PMID- 20012669 TI - Platelets and viruses: an ambivalent relationship. AB - Thrombocytopenia is a frequent complication of viral infections providing evidence that interaction of platelets with viruses is an important pathophysiological phenomenon. Multiple mechanisms are involved depending on the nature of the viruses involved. These include immunological platelet destruction, inappropriate platelet activation and consumption, and impaired megakaryopoiesis. Viruses bind platelets through specific receptors and identified ligands, which lead to mutual alterations of both the platelet host and the viral aggressor. We have shown that HIV-1 viruses are internalized specifically in platelets and megakaryocytes, where they can be either sheltered, unaltered (with potential transfer of the viruses into target organs), or come in contact with platelet secretory products leading to virus destruction and facilitated platelet clearance. In this issue, we have reviewed the various pathways that platelets use in order to interact with viruses, HIV and others. This review also shows that more work is still needed to precisely identify platelet roles in viral infections, and to answer the challenge of viral safety in platelet transfusion. PMID- 20012670 TI - Meningioma of the cerebellopontine angle in identical twins: a case report. AB - Meningiomas in identical twins are extremely rare. To our knowledge, only one previous report of meningiomas in identical twins has been published. We present identical twin sisters with meningiomas. The tumors were located at a similar, but not a common, position (the cerebellopontine angle) in both twins. Histologically, both tumors were diagnosed as meningothelial meningiomas with an angiomatous component. Immunohistochemically, the Ki-67 indices in the two cases were 1.0 and 1.1, and the p53 positive rates were 0.2 and 0.9. The specimens in both cases were reactive to neurofibromin 2 (NF2). A comparative genomic hybridization (CGH) assay revealed an aberration in the long arm of chromosome X, but no aberrations in the long arm of chromosome 22 in either case. These results strongly suggest that genetic aberrations other than NF2 are associated with tumorigenesis in some types of sporadic meningiomas. PMID- 20012671 TI - Interactions between anti-ErbB2 antibody A21 and the ErbB2 extracellular domain provide a basis for improving A21 affinity. AB - Anti-ErbB2 antibodies are well researched for the therapy of ErbB2-overexpressing tumors. The therapeutic potential and efficacy of these antibodies are closely related to their affinities to ErbB2. Previously we reported that an anti-ErbB2 antibody A21 targeting a conformational epitope comprising several loops in ErbB2 extracellular subdomain I and II could inhibit the proliferation of ErbB2 overexpressing cancer cells in vitro and in vivo. Here we found that another structureless and non-conserved loop in subdomain I of ErbB2 extracellular domain (ECD) was important for binding to A21, and then the antigen-contact sites on A21 were determined by site-directed mutation. The loop was constructed by molecular modeling, and a new model of A21-ErbB2 complex was generated by docking using the crystal structure of the scfv A21 and the model of ErbB2 ECD with the loop built. Based on the complex model, computational design for A21 affinity improvement was performed to enhance its affinity to ErbB2. Two mutants with about 1.7-fold improvement in affinity were obtained. Our study provided a rational molecular basis for affinity improvement and mechanism investigation of A21. PMID- 20012672 TI - Arabidopsis protochlorophyllide oxidoreductase A (PORA) restores bulk chlorophyll synthesis and normal development to a porB porC double mutant. AB - In angiosperms the strictly light-dependent reduction of protochlorophyllide to chlorophyllide is catalyzed by NADPH:protochlorophyllide oxidoreductase (POR). The Arabidopsis thaliana genome encodes three structurally related but differentially regulated POR genes, PORA, PORB and PORC. PORA is expressed primarily early in development-during etiolation, germination and greening. In contrast, PORB and PORC are not only expressed during seedling development but also throughout the later life of the plant, during which they are responsible for bulk chlorophyll synthesis. The Arabidopsis porB-1 porC-1 mutant displays a severe xantha (highly chlorophyll-deficient) phenotype characterized by smaller prolamellar bodies in etioplasts and decreased thylakoid stacking in chloroplasts. Here we have demonstrated the ability of an ectopic PORA overexpression construct to restore prolamellar body formation in the porB-1 porC 1 double mutant background. In response to illumination, light-dependent chlorophyll production, thylakoid stacking and photomorphogenesis are also restored in PORA-overexpressing porB-1 porC-1 seedlings and adult plants. An Arabidopsis porB-1 porC-1 double mutant can therefore be functionally rescued by the addition of ectopically expressed PORA, which suffices in the absence of either PORB or PORC to direct bulk chlorophyll synthesis and normal plant development. PMID- 20012674 TI - Population genetic structure of the oriental fruit fly, Bactrocera dorsalis (Hendel) (Diptera: Tephritidae) from Yunnan province (China) and nearby sites across the border. AB - Bactrocera dorsalis (Diptera: Tephritidae) is an important pest for many tropical and subtropical fruits. The fly is probably introduced in Yunnan, a southwestern province of China that shares borders with Vietnam, Laos and Myanmar. Depending on local environmental conditions, this species occurs either only in the most favorable seasons or year-round. To infer the genetic diversity and structure of the fly in the region, and to understand the relationships between the flies of year-round and seasonal areas, we analyzed 304 individuals from 14 populations using the mitochondrial cytochrome oxidase I gene (COI). The sampled populations were structured into four groups, probably isolated by the main natural barriers in Yunnan such as mountain ranges and rivers. Our data suggest either that B. dorsalis in Yunnan originated from multiple introductions events, even if the source populations still need to be identified; or that Yunnan is a natural origin of this species (i.e., that it is not invasive there). Finally, we found some evidences that the seasonal populations were colonized from nearby year round populations. PMID- 20012673 TI - Luminescence detection of SNARE-SNARE interaction in Arabidopsis protoplasts. AB - Membrane associated proteins SNAREs (soluble N-ethylmaleimide-sensitive fusion protein attachment protein receptors) provide the minimal fusion machinery necessary for cellular vesicles to fuse to target organelle membranes in eukaryotic cells. Despite the conserved nature of the fusion machinery in all eukaryotes, it still remains challenging to identify functional SNARE pairs in higher plants. We developed a method based on a split-luciferase complementation assay for detecting changes in SNARE-SNARE interaction by luminescence within Arabidopsis protoplasts that express recombinant proteins at physiological levels in 96-well plates. The reliability of the assay was confirmed by three experiments. First, reduction of the SNARE-SNARE interaction caused by a single amino acid substitution adjacent to the SNARE motif in endosome-localized AtVAM3/SYP22 (syntaxin of plant 22) was detected by a reduction of luminescence. Second, reduction of the interaction between plasma-membrane localized SYP121 and VAMP722 in response to sodium azide was detected in real-time. Third, the results of 21 SNARE pairs investigated by this method largely agreed with the results from previously reported co-immunoprecipitation assays. Using the method, we newly identified the interaction between SYP121 and VAMP722 was significantly increased when the protoplasts were incubated in the light. Microscopic observation of transgenic Arabidopsis expressing GFP-SYP121 (green fluorescent protein tagged SYP121) from its own promoter suggested that the plasma-membrane localization of GFP-SYP121 is maintained by light. These suggested that the vesicle trafficking pathway mediated by SYP121 might be regulated by light in Arabidopsis. In general, this article demonstrated the method that can generate new biological insight of the SNARE protein interactions in plant cells. PMID- 20012675 TI - Diversity of the volatile organic compounds emitted by 55 species of tropical trees: a survey in French Guiana. AB - Volatile organic compounds (VOCs) are produced by a broad range of organisms, from bacteria to mammals, and they represent a vast chemical diversity. In plants, one of the preeminent roles of VOCs is their repellent or cytotoxic activity, which helps the plant deter its predators. Most studies on VOCs emitted by vegetative parts have been conducted in model plant species, and little is known about patterns of VOC emissions in diverse plant communities. We conducted a survey of the VOCs released immediately after mechanical damage of the bark and the leaves of 195 individual trees belonging to 55 tropical tree species in a lowland rainforest of French Guiana. We discovered a remarkably high chemical diversity, with 264 distinct VOCs and a mean of 37 compounds per species. Two monoterpenes (alpha-pinene and limonene) and two sesquiterpenes (beta caryophyllene and alpha-copaene), which are known to have cytotoxic and deterrent effects, were the most frequent compounds in the sampled species. As has been established for floral scents, the blend of VOCs is largely species-specific and could be used to discriminate among 43 of the 55 sampled species. The species with the most diverse blends were found in the Sapindales, Laurales, and Magnoliales, indicating that VOC diversity is not uniformly distributed among tropical species. Interspecific variation in chemical diversity was caused mostly by variation in sesquiterpenes. This study emphasizes three aspects of VOC emission by tropical tree species: the species-specificity of the mixtures, the importance of sesquiterpenes, and the wide-ranging complexity of the mixtures. PMID- 20012676 TI - Identification, synthesis and field testing of (3Z,6Z,9Z)-3,6,9-henicosatriene, a second bioactive component of the sex pheromone of the autumn gum moth, Mnesampela privata. AB - The sex pheromone of Mnesampela privata, an endemic pest of Eucalyptus plantations in Australia, was previously identified as a single bioactive compound, (3Z,6Z,9Z)-3,6,9-nonadecatriene (C19 triene). Initial field testing of lures containing 1 mg, 5 mg or 10 mg of C19 triene (>98% purity) caught no or very few male M. privata. (3Z,6Z,9Z)-3,6,9-Henicosatriene (C21 triene) was identified as an additional minor pheromone component in abdominal tip extracts of M. privata females from Tasmania. Levels of both compounds extracted from individual females varied greatly, but the ratio was relatively constant at 33:1 C19:C21 trienes. Electroantennograms (EAG) of synthetic C21 triene with male M. privata gave positive but consistently lower responses than elicited by the C19 triene. Field tests showed that the addition of 1-6% C21 triene to 1 mg C19 triene significantly increased trap catch and the detection of M. privata in plantations. Traps baited with a 16:1 ratio caught significantly more moths than those baited with a ratio approximating that of females. PMID- 20012678 TI - Early interests and joint engagement in typical development, autism, and Down syndrome. AB - This study examines how spontaneous interests in people and in objects relate to joint engagement in typically developing toddlers and young children with autism or Down syndrome. Ratings of interests were made repeatedly during intermissions in a laboratory-based protocol focused on caregiver-child interactions. Interests were moderated by diagnosis and relatively stable across intermissions. In autism, interest in people tended to be low and to decline rapidly, and the balance of interests favored familiar objects over people. Lower interest in people and in unfamiliar objects was associated with less coordinated joint engagement and with less steep developmental trajectories for symbol-infused joint engagement. These findings suggest that variations in interests may contribute to differences in the child's engagement during social interactions that facilitate the acquisition of language. PMID- 20012677 TI - Household exposure to secondhand smoke is associated with decreased physical and mental health of mothers in the USA. AB - Secondhand smoke is one of the most common toxic environmental exposures to children, and maternal health problems also have substantial negative effects on children. We are unaware of any studies examining the association of living with smokers and maternal health. To investigate whether non-smoking mothers who live with smokers have worse physical and mental health than non-smoking mothers who live in homes without smokers. Nationally representative data from the 2000-2004 Medical Expenditure Panel Survey were used. The health of non-smoking mothers with children <18 years (n = 18,810) was assessed, comparing those living with one or more smokers (n = 3,344) to those living in households with no adult smokers (n = 14,836). Associations between maternal health, household smoking, and maternal age, race/ethnicity, and marital, educational, poverty and employment status were examined in bivariable and multivariable analyses using SUDAAN software to adjust for the complex sampling design. Scores on the Medical Outcomes Short Form-12 (SF-12) Physical Component Scale (PCS) and Mental Component Scale (MCS) were used to assess maternal health. About 79.2% of mothers in the USA are non-smokers and 17.4% of them live with >=1 adult smokers: 14.2% with 1 and 3.2% with >=2 smokers. Among non-smoking mothers, the mean MCS score is 50.5 and mean PCS is 52.9. The presence of an adult smoker and increasing number of smokers in the home are both negatively associated with MCS and PCS scores in bivariable analyses (P < 0.001 for each). Non-smoking mothers with at least one smoker in the household had an 11% (95% CI = 0.80-0.99) lower odds of scoring at or above the mean MCS score and a 19% (95% CI = 0.73-0.90) lower odds of scoring at or above the mean PCS score compared to non-smoking mothers with no smokers in the household. There is an evidence of a dose response relationship with increasing number of smokers in the household for PCS (P < 0.001). These findings demonstrate a previously unrecognized child health risk: living with smokers is independently associated with worse physical and mental health among non-smoking mothers. PMID- 20012679 TI - Identification of recombination between subgenotypes IA and IB of hepatitis A virus. AB - Co-circulation of subgenotypes IA and IB of hepatitis A virus (HAV) has been reported in South Africa, South America, Europe, and the United States. In this study, phylogenetic and recombination analyses were performed for the first time on 31 complete HAV genomes from infected humans and simians. Three potentially significant intra-genotypic recombination events (I-III) were identified by recombination detection analysis. Recombination events I and II occurred between the lineages represented, respectively, by the Japanese isolate AH2 (AB020565, subgenotype IA) and the North African isolate MBB (M20273, subgenotype IB), giving rise to the recombinant Uruguayan isolate HAV5 (EU131373). Recombination event III occurred between the lineages represented, respectively, by the North African isolate MBB (M20273, subgenotype IB) and the German isolate GBM (X75215, subgenotype IA), resulting in the Italian isolate FG (X83302). The findings demonstrate that humans can be co-infected with different HAV subgenotypes and provide valuable hints for future research on HAV diversity. PMID- 20012680 TI - Full-length hepatitis B virus sequences from naive patients with fluctuation of viral load during ADV monotherapy. AB - The reasons for adefovir dipivoxil (ADV) treatment failures appear diverse. Few studies have reported full-length hepatitis B virus (HBV) genome in patients with ADV treatment failures. The patients were from a phase III clinical trial that investigated the antiviral response to ADV in China. Seven patients had increase in HBV-DNA (>1 log(10) copies/ml above on-treatment nadir) at week 52. The serum HBV-DNA levels were above 10(4)copies/ml at week 92 in four of them. Sixteen full length HBV genomes from the four patients at four time points were sequenced using cloning sequencing method. The frequency of substitutions at week 52 was higher than at weeks 28(16 wt) and 92(80). HBV-DNA reduction was correlated negatively with the frequency of substitutions at the three time points. No published ADV-resistant mutations were detected. The mutations, including substitutions in immunogenic epitopes and conserved sites of the polymerase gene, were frequent during ADV treatment. Amino acid deletions in X gene and basal core promoter/pre-core mutations appeared before or during ADV treatment. The substitutions in immunogenic epitopes (mainly of the surface gene) and conserved sites of the polymerase gene other than ADV-resistant mutations may have influenced antiviral efficacy in the study. More potent antiviral drugs may be important to rescue individual patients and for public health safety. It is needed to study how these substitutions influence HBV replication, disease progression, and antiviral treatment efficacy. PMID- 20012681 TI - Genetic comparisons between lentogenic Newcastle disease virus isolated from waterfowl and velogenic variants. AB - Avirulent Newcastle disease viruses (NDV) harbored by waterfowl have the potential to become virulent after transmission to and circulation within chicken populations. In order to investigate how virulent viruses are selected from an avirulent background, we compared the complete sequences of the avirulent NDV isolate Goose/Alaska/415/91 and its virulent variant strain 9a5b, which was obtained by nine and five passages in the chick air sac and brain, respectively. Seven amino acid substitutions were detected in the M, F, and HN proteins. Two were detected between variants 9a3b and 9a5b (128P to H and 495E to K in HN protein) that were passed through the brain. Pathogenicity determined by the MDT and IVPI tests also differed between 9a3b and 9a5b. These results suggest that in addition to the F cleavage site sequence, these two amino acids in HN protein are also related to the pathogenicity of NDV in chickens. PMID- 20012682 TI - The neural bases of word encoding and retrieval: A fMRI-guided transcranial magnetic stimulation study. AB - There is evidence that the human prefrontal cortex is asymmetrically involved in long-term episodic memory processing. Moreover, abstract and concrete words processing has been reported to differentially involve prefrontal and parietal areas. We implemented a two-stages functional magnetic resonance imaging (fMRI) repetitive transcranial magnetic stimulation (rTMS) paradigm to investigate the role of the dorsolateral prefrontal cortices (DLPFCs) and parietal cortices (PARCs) in encoding and retrieval of abstract and concrete words. Using this paradigm we could select areas to be stimulated on the basis of single-subject (SS) anatomical and functional data, investigating the usefulness of this integration approach. With respect to fMRI, abstract and concrete words differed only for a greater left fusiform gyrus activation for concrete words. In turn, significant rTMS effects were found, but only for the retrieval of abstract words. Consistent with previous findings, repetitive stimulation of the right DLPFC had a specific impact on episodic retrieval. Memory retrieval performance was also disrupted when rTMS was applied to the left PARC. Finally, we found a significant positive correlation between the effect sizes of SS right PARC activations for abstract word retrieval and the consequent rTMS interference effects. Taken together these data provide for the first time evidence that also the PARC has a necessary role in episodic retrieval of abstract words. Importantly, from a methodological perspective, our data demonstrate that fMRI guided rTMS with a SS approach provides a powerful tool to investigate the neural underpinnings of cognitive functions. PMID- 20012683 TI - Spontaneously developed tail swellings (SDTS) influence the accuracy of the hypo osmotic swelling test (HOS-test) in determining membrane integrity and viability of human spermatozoa. AB - PURPOSE: To investigate the prevalence of spontaneously developed tail swellings (SDTS) in human sperm samples that are commonly encountered in the laboratory, and their influence on the hypo-osmotic swelling test (HOS-test). METHODS: Ejaculated, epididymal, and testicular sperm were evaluated for SDTS. Further, HOS-test scores were compared with those of vital stains using column washed sperm maintained in the laboratory. RESULTS: SDTS, at <10%, was present in all types of sperm samples. The highest and lowest occurrences of SDTS were found in cryopreserved sperm, and column-washed sperm respectively. SDTS can inflate the HOS-test score, and so lower the accuracy of the HOS-test. However, the HOS-test efficiency can be improved by assessing SDTS in the sample. CONCLUSION: HOS-test and vital stain cannot be used interchangeably in all circumstances for sperm viability determination. The accuracy of the HOS-test can be enhanced by incorporating SDTS as a correction factor. PMID- 20012684 TI - The shape of the sperm midpiece in intracytoplasmic morphologically selected sperm injection relates sperm centrosomal function. AB - PURPOSE: To evaluate whether the morphology of the sperm midpiece observed by high magnification microscopy relates to sperm centrosomal function. METHODS: Sperm selected by conventional microscopy were defined as controls. By high magnification microscopy, sperm with straight midpieces were defined as Group 1, while those with tapering midpieces were defined as Group 2. Heterologous ICSI of human sperm into bovine oocytes was used to assess human sperm centrosomal function and analysis of sperm aster formation. RESULTS: The total rate of sperm aster formation was 80.5% in Group 1, which was significantly higher (P < 0.05) than the rate of 33.3% seen for Group 2. Furthermore, sperm aster formation rates tended to be higher for Group 1 than for the controls. CONCLUSIONS: This study demonstrates improvement of sperm aster formation rates by selecting sperm on the basis of midpiece morphology. The injection of selected sperm bearing morphologically straight midpieces may contribute to improved expression of sperm centrosomal function, providing a positive effect on fertilization after ICSI. PMID- 20012685 TI - Sperm DNA damage in male infertility: etiologies, assays, and outcomes. AB - Male factor infertility is the sole cause of infertility in approximately 20% of infertile couples, with an additional 30% to 40% secondary to both male and female factors. Current means of evaluation of male factor infertility remains routine semen analysis including seminal volume, pH, sperm concentration, motility, and morphology. However, approximately 15% of patients with male factor infertility have a normal semen analysis and a definitive diagnosis of male infertility often cannot be made as a result of routine semen analysis. Attention has focused on the role of sperm nuclear DNA integrity in male factor infertility. Here we review the structure of human sperm chromatin, the etiology and mechanisms of sperm DNA damage, current tests available to assess sperm DNA integrity, and effect of sperm DNA integrity on reproductive outcomes. PMID- 20012686 TI - Pass-fail grading: laying the foundation for self-regulated learning. AB - Traditionally, medical schools have tended to make assumptions that students will "automatically" engage in self-education effectively after graduation and subsequent training in residency and fellowships. In reality, the majority of medical graduates out in practice feel unprepared for learning on their own. Many medical schools are now adopting strategies and pedagogies to help students become self-regulating learners. Along with these changes in practices and pedagogy, many schools are eliminating a cornerstone of extrinsic motivation: discriminating grades. To study the effects of the switch from discriminating to pass-fail grading in the second year of medical school, we compared internal and external assessments and evaluations for a second-year class with a discriminating grading scale (Honors, High Pass, Pass, Fail) and for a second year class with a pass-fail grading scale. Of the measures we compared (MCATs, GPAs, means on second-year examinations, USMLE Step 1 scores, residency placement, in which there were no statistically significant changes), the only statistically significant decreases (lower performance with pass fail) were found in two of the second-year courses. Performance in one other course also improved significantly. Pass-fail grading can meet several important intended outcomes, including "leveling the playing field" for incoming students with different academic backgrounds, reducing competition and fostering collaboration among members of a class, more time for extracurricular interests and personal activities. Pass-fail grading also reduces competition and supports collaboration, and fosters intrinsic motivation, which is key to self-regulated, lifelong learning. PMID- 20012688 TI - Comparison of ionized calcium-binding adapter molecule 1-immunoreactive microglia in the spinal cord between young adult and aged dogs. AB - Microglia are main form of active immune defense, and they are constantly moving and analyzing the CNS for damaged neurons and infectious agents. In this study, we compared microglia in the spinal cord of the young adult (1-2 years old) and aged (10-12 years old) German Shepherd dogs via immunohistochemistry and western blot analysis for ionized calcium-binding adapter molecule 1 (Iba-1), a microglial marker. In addition, we also observed the interferon-gamma (IFN gamma), a pro-inflammatory cytokine, and interleukin-1beta (IL-1beta), produced by activated microglia/macrophage, protein levels in these groups. At first, we found that neuronal nuclei (NeuN, a neuronal marker)-immunoreactive neurons were distributed throughout the grey mate of the spinal cord, and there were no significant differences between the adult and aged groups. Most of Iba-1 immunoreactive microglia were morphologically ramified microglia (resting form) in the adult group, while some Iba-1-immunoreactive microglia were morphologically activated microglia in the aged group. In western blot analysis, Iba-1, IFN-gamma and IL-1beta expression were increased in the aged group. This result may be associated with age-dependent changes in the spinal cord. PMID- 20012690 TI - Milk yield and associated economic losses in quarters with subclinical mastitis due to Staphylococcus aureus in Ethiopian crossbred dairy cows. AB - The objective of the study was to estimate the prevalence and losses associated with subclinical mastitis (SCM) caused by Staphylococcus aureus in Ethiopian crossbred dairy cows. A split-udder trial was carried out to determine milk yield losses in udder quarter with S. aureus-caused SCM. Each quarter of the study cows was examined using the California Mastitis Test (CMT) and quarter milk production was measured over a period of 8 days. Milk yield losses for CMT positive quarters were estimated by comparing production of quarters with CMT score 0. Mean milk yield for uninfected healthy quarters was 1.66 kg per milking (95% CI, 1.66-1.55 kg per milking), and the rate of milk reduction for quarters with CMT scores of 1+, 2+, and 3+ was 25%, 33%, and 48%, respectively. Economic losses at different farm-size levels were calculated by multiplying the prevalence of CMT scores with milk yield losses associated with respective CMT scores. In Debre Ziet dairy herds, a quarter with SCM due to S. aureus lost an average of 34.5% of its potential milk production while the total milk yield loss per cow was estimated at 6.8%. Losses were highest in large-scale (13%) farms and lowest (3.7%) in small-scale. Based on the prevalence, the overall financial loss for each cow per lactation was 984.64 Eth Birr (US$78.65) and losses in large farms (1,882.40 Eth Birr or US$150.35) were over 3.5 times the loss in small-size farms. These figures possibly underestimate the potential benefits of mastitis control program as they do not include other direct and indirect costs. PMID- 20012692 TI - ATP transport in saccular cerebral aneurysms at arterial bends. AB - ATP acts as an extracellular signaling molecule in purinergic signaling that regulates vascular tone. ATP binds purinergic P2 nucleotide receptors on endothelial cells. Understanding the mass transport of ATP to endothelial cells by blood flow is thus important to predict functional changes in aneurysmal walls. While some clinical observations indicate a difference of wall pathology between ruptured and unruptured aneurysms, no study has focused on the mass transport in aneurysms. We investigated the characteristics of ATP concentration at aneurysmal wall using a numerical model of ATP transport in aneurysms formed at arterial bends. The magnitude of ATP concentration at the aneurysmal wall was significantly smaller than that at the arterial wall. In particular, significantly low concentration was predicted at the proximal side of the aneurysmal sac. A strong correlation was revealed between the inflow flux at the aneurysmal neck and the resultant concentration at the aneurysmal wall. PMID- 20012691 TI - Prevalence and associated risk factors of mycobacterial infections in slaughter pigs from Mubende district in Uganda. AB - To date, the public health relevance of mycobacterial infections in pigs is not well investigated despite high risk of infection. Recently, there has been a documented increase in opportunistic infections and risk of acquiring opportunistic mycobacterial infections in HIV/AIDS patients in Mubende district; unfortunately, there has been no published information on the epidemiology of mycobacterial infections in this area. This study was carried out between September 2008 and February 2009. Investigations were done to assess the prevalence and associated risk factors of mycobacterial infections in slaughtered pigs in Mubende district of Uganda. A total of 997 pigs (53.7% male and 46.3% female) from 31 different slaughterhouses were examined for the presence of lesions compatible with TB and mycobacterial infections. Pathologic tissue specimens were collected for culturing and isolation of mycobacteria. A cross sectional technique was used based on convenient visits to slaughterhouses but random selection of individual slaughtered pigs for a detailed post-mortem inspection on a daily basis. The results reflected a 9.3% and 3.1% (95% CI) prevalence of Mycobacterium species based on necropsy examinations and culture isolation, respectively. The highest prevalence of mycobacterial infection was recorded in Buwekula County (the mixed agro-zone) whilst the lowest was in Kassanda County (pastoral zone). A multivariable logistical regression analysis identified age (P < or = 0.001) and sex (P < or = 0.05) as risk factors for mycobacterial infections in pigs. Post-estimation statistics of the regression model evaluation and validation fit it well into the data (HL, chi (2) = 5.9; P = 0.69 for necropsy, HL chi (2) = 2.9; P = 0.94 for culturing). This study documented a high prevalence of mycobacterial infections in slaughter pigs in Mubende district. The fact that pigs and human often share common housing and environment poses a high risk of zoonotic transmission. This then warrants further molecular investigation to identify the specific Mycobacterium species and their public health importance in this area. PMID- 20012693 TI - Multiscale analysis of heart rate variability: a comparison of different complexity measures. AB - Heart rate variability (HRV) is an important dynamical variable of the cardiovascular function. There have been numerous efforts to determine whether HRV dynamics are chaotic or random, and whether certain complexity measures are capable of distinguishing healthy subjects from patients with certain cardiac disease. In this study, we employ a new multiscale complexity measure, the scale dependent Lyapunov exponent (SDLE), to characterize the relative importance of nonlinear, chaotic, and stochastic dynamics in HRV of healthy, congestive heart failure (CHF), and atrial fibrillation subjects. We show that while HRV data of all these three types are mostly stochastic, the stochasticity is different among the three groups. Furthermore, we show that for the purpose of distinguishing healthy subjects from patients with CHF, features derived from SDLE are more effective than other complexity measures such as the Hurst parameter, the sample entropy, and the multiscale entropy. PMID- 20012687 TI - Heterogeneity of reward mechanisms. AB - The finding that many drugs that have abuse potential and other natural stimuli such as food or sexual activity cause similar chemical changes in the brain, an increase in extracellular dopamine (DA) in the shell of the nucleus accumbens (NAccS), indicated some time ago that the reward mechanism is at least very similar for all stimuli and that the mechanism is relatively simple. The presently available information shows that the mechanisms involved are more complex and have multiple elements. Multiple brain regions, multiple receptors, multiple distinct neurons, multiple transmitters, multiple transporters, circuits, peptides, proteins, metabolism of transmitters, and phosphorylation, all participate in reward mechanisms. The system is variable, is changed during development, is sex-dependent, and is influenced by genetic differences. Not all of the elements participate in the reward of all stimuli. Different set of mechanisms are involved in the reward of different drugs of abuse, yet different mechanisms in the reward of natural stimuli such as food or sexual activity; thus there are different systems that distinguish different stimuli. Separate functions of the reward system such as anticipation, evaluation, consummation and identification; all contain function-specific elements. The level of the stimulus also influences the participation of the elements of the reward system, there are possible reactions to even below threshold stimuli, and excessive stimuli can change reward to aversion involving parts of the system. Learning and memory of past reward is an important integral element of reward and addictive behavior. Many of the reward elements are altered by repeated or chronic stimuli, and chronic exposure to one drug is likely to alter the response to another stimulus. To evaluate and identify the reward stimulus thus requires heterogeneity of the reward components in the brain. PMID- 20012694 TI - A numerical model of the respiratory modulation of pulmonary shunt and PaO2 oscillations for acute lung injury. AB - It is an accepted hypothesis that the amplitude of the respiratory-related oscillations of arterial partial pressure of oxygen (DeltaPaO2) is primarily modulated by fluctuations of pulmonary shunt (Deltas), the latter generated mainly by cyclic alveolar collapse/reopening, when present. A better understanding of the relationship between DeltaPaO2, Deltas, and cyclic alveolar collapse/reopening can have clinical relevance for minimizing the severe lung damage that the latter can cause, for example during mechanical ventilation (MV) of patients with acute lung injury (ALI). To this aim, we numerically simulated the effect of such a relationship on an animal model of ALI under MV, using a combination of a model of lung gas exchange during tidal ventilation with a model of time dependence of shunt on alveolar collapse/opening. The results showed that: (a) the model could adequately replicate published experimental results regarding the complex dependence of DeltaPaO2 on respiratory frequency, driving pressure (DeltaP), and positive end-expiratory pressure (PEEP), while simpler models could not; (b) such a replication strongly depends on the value of the model parameters, especially of the speed of alveolar collapse/reopening; (c) the relationship between DeltaPaO2 and Deltas was overall markedly nonlinear, but approximately linear for PEEP>or=6 cmH2O, with very large DeltaPaO2 associated with relatively small Deltas. PMID- 20012695 TI - Fingertip digital thermal monitoring: a fingerprint for cardiovascular disease? PMID- 20012696 TI - An electrically active microneedle array for electroporation. AB - We have designed and fabricated a microneedle array with electrical functionality with the final goal of electroporating skin's epidermal cells to increase their transfection by DNA vaccines. The microneedle array was made of polymethylmethacrylate (PMMA) by micromolding technology from a polydimethylsiloxane (PDMS) mold, followed by metal deposition, patterning using laser ablation, and electrodeposition. This microneedle array possessed sufficient mechanical strength to penetrate human skin in vivo and was also able to electroporate both red blood cells and human prostate cancer cells as an in vitro model to demonstrate cell membrane permeabilization. A computational model to predict the effective volume for electroporation with respect to applied voltages was constructed from finite element simulation. This study demonstrates the mechanical and electrical functionalities of the first MEMS-fabricated microneedle array for electroporation, designed for DNA vaccine delivery. PMID- 20012697 TI - The risk for breast cancer is not evidently increased in women with hyperprolactinemia. AB - The question has been raised whether hyperprolactinemia in humans is associated with an excess risk for breast cancer. We aimed to assess the risk of breast cancer in a previously defined large cohort of patients treated for idiopathic hyperprolactinemia or prolactinomas. Based on the pattern of drug prescriptions we identified 11,314 subjects in the PHARMO network with at least one dispensing of dopamine agonists between 1996 and 2006. Of these, 1,607 subjects were considered to have dopamine agonist-treated hyperprolactinemia based on the prescribing pattern. For the present analysis, we included only women (n = 1,342). Patients with breast cancer were identified by hospital discharge codes. Data on breast cancer incidence in the Netherlands were derived from the Dutch cancer registry. Standardized mortality ratio (SMR) was the measure of outcome to assess the association between hyperprolactinemia and breast cancer. The 1,342 patients accounted for a total of 6,576 person years. Eight patients with breast cancer during follow-up were identified. Indirect standardization with incidence proportions from the general Dutch population revealed a 7.47 expected cases. The calculated SMR for breast cancer risk in patients treated hyperprolactinemia was 1.07 (95% confidence interval 0.50-2.03). In conclusion, there is no clear evidence for increased breast cancer risk in female patients treated for either idiopathic hyperprolactinemia or prolactinomas. The uncertainty about the exact risk that is due to the relatively low number of breast cancer cases, should be overcome by pooling results in a future meta-analysis. PMID- 20012698 TI - Evaluation of antioxidant systems in pituitary-adrenal axis diseases. AB - The role of adrenal steroids in antioxidant regulation is not known. Previously, we demonstrated some Coenzyme Q(10) (CoQ(10)) alterations in pituitary diseases, which can induce complex pictures due to alterations of different endocrine axes. Therefore we determined CoQ(10) and Total Antioxidant Capacity (TAC) in pituitary dependent adrenal diseases: 6 subjects with ACTH-dependent adrenal hyperplasia (AH); 19 with secondary isolated hypoadrenalism (IH), 19 with associated hypothyroidism (multiple pituitary deficiencies, MPH). CoQ(10) was assayed by HPLC; TAC by the system metmyoglobin-H(2)O(2), which, interacting with the chromogenous 2,2(I)-azinobis-(3-ethylbenzothiazoline-6-sulphonate), generates a spectroscopically revealed radical compound after a latency time (Lag) proportional to the antioxidant content. CoQ(10) levels were significantly lower in IH than AH and MPH, with a similar trend when adjusted for cholesterol. Also TAC was lower in IH than in AH and MPH, suggesting that adrenal hormones can influence antioxidants. However, since thyroid hormones modulate CoQ(10) levels and metabolism, when thyroid deficiency coexists it seems to play a prevalent influence. PMID- 20012699 TI - A heritable predisposition to pituitary tumors. AB - Pituitary tumors are prevalent in the general population, with a frequency of nearly 1 in 5. The cause of most pituitary tumors remains unknown, although a genetic contribution is recognized for some. We analyzed the Utah Population Data Base (UPDB), a resource combining a computerized genealogy of the Utah population with a statewide tumor registry, to investigate familial clustering of pituitary tumors. We analyzed the genetic relationships among 741 individuals diagnosed with benign or malignant pituitary tumors who had Utah genealogy data. To test for evidence of genetic contribution to predisposition, we compared average relatedness between all pairs of individuals with pituitary tumors with the expected relatedness in this population. We also estimated relative risks (RRs) for pituitary tumors in close and distant relatives of cases by comparing observed and expected numbers of cases among relatives. Relative risks for first- and third-degree relatives were significantly elevated (RR = 2.83 and 1.63, respectively), while relative risk for second-degree relatives was not significantly different from 1.0 (RR = 0.83). The average pairwise relatedness of pituitary tumor cases was significantly higher than expected, even when close relationships were ignored. The significantly elevated risks to relatives as well as the significant excess distant relatedness observed in cases provide strong support for a genetic contribution to predisposition to pituitary tumors. Multiple high-risk pedigrees can be identified in the UPDB, and study of such pedigrees might allow identification of the gene(s) responsible for our observations. Recognizing genetic contribution to the disease may also help with counseling family members of affected individuals. PMID- 20012700 TI - Clinically-staged T3N0 rectal cancer: is preoperative chemoradiotherapy the optimal treatment? AB - BACKGROUND: Preoperative chemoradiotherapy has been widely adopted as the standard of care for stage II-III rectal cancers. However, patients with T3N0 lesions had been shown to have a better prognosis than other categories of locally advanced tumor. Thus, neoadjuvant chemoradiation is likely to be overtreatment in this subgroup of patients. Nevertheless, the low accuracy rate of preoperative staging techniques for detection of node-negative tumors does not allow to check this hypothesis. We analyzed a group of patients with cT3N0 low rectal cancer who underwent neoadjuvant chemoradiotherapy with the purpose of evaluating the incidence of metastatic nodes in the resected specimens. METHODS: Between January 2002 and February 2008, 100 patients with low rectal cancer underwent clinical staging by means of endorectal ultrasound, computed tomography, positron emission tomography, and magnetic resonance imaging. All patients received preoperative 5-fluorouracil-based chemoradiotherapy and surgical resection with curative aim. RESULTS: Of 100 patients with locally advanced rectal cancer, 32 were clinically staged as T3N0M0. Pathological analysis showed the presence of lymph node metastases in nine patients (28%) (node-positive group). In the remaining 23 cases, clinical N stage was confirmed at pathology (node-negative group). Node-positive and node-negative groups differ only in the number of ypT3 tumors (P < .01). CONCLUSIONS: Our results indicate that immediate surgery for patients with cT3N0 rectal cancer represents an undertreatment risk in at least 28% of cases, making necessary the use of postoperative chemoradiotherapy. Preoperative chemoradiotherapy should be the therapy of choice on the grounds of the principle that overtreatment is less hazardous than undertreatment for cT3N0 rectal cancers. PMID- 20012701 TI - Degree of tumor FDG uptake correlates with proliferation index in triple negative breast cancer. AB - PURPOSE: 2-Deoxy-2-[F-18]fluoro-D-glucose (FDG) uptake may be a useful surrogate marker for proliferation index, but the correlation has not always been clear cut. Previous research by our group suggests that FDG-positron emission tomography (PET) is sensitive in detecting triple negative breast cancer. We therefore performed a pilot study to test if FDG uptake correlated with proliferation index in women with triple negative cancer. PROCEDURES: To determine whether proliferation index correlates with metabolic uptake of FDG in women with triple negative breast cancer, we performed a retrospective analysis correlating %Ki67 nuclear stain with tumor maximum standardized uptake values (SUVmax) in a group of 41 women, 22 with triple negative and 19 with non-triple negative breast cancer. RESULTS: As expected, [18F]-PET imaging was significantly more sensitive in detecting triple negative breast cancer than non-triple negative breast cancer, 95.5% vs 68.4% (p = 0.036). In general, SUVmax and %Ki67 nuclear stain values rise as histologic grade worsens. Histologic grade of triple negative breast cancer was more often poorly differentiated than non-triple negative cancer (p = 0.001). SUVmax correlated with %Ki67 nuclear staining in our entire cohort (spearman correlation = 0.485, p = 0.002). Moreover, this significant correlation appeared to be driven primarily by a subset of women with triple negative cancer (spearman correlation = 0.497, p = 0.019). CONCLUSIONS: Degree of tumor FDG uptake correlated significantly with proliferation index in women with triple negative breast cancer suggesting a potential role of FDG-PET in treatment response monitoring for this group of women. Future studies are necessary to define the role of PET imaging as a non-invasive means to monitor breast cancer treatment response in the neoadjuvant setting. PMID- 20012702 TI - Emergency contraception for newly arrested women: evidence for an unrecognized public health opportunity. AB - Incarceration affords an opportunity to provide health care to populations with limited access to care. Women in this population are at high risk for experiencing unintended pregnancies. It is not known what proportion of these women engage in unprotected intercourse in the days prior to incarceration and therefore may benefit from being offered emergency contraception upon their arrest to decrease their risk of unintended pregnancies. We sought to describe the proportion and characteristics of newly arrested women who are eligible for and interested in taking emergency contraception by conducting a cross-sectional study in an urban county jail booking facility. A 63-item survey was administered to women ages 18-44 within 24 h of being arrested in San Francisco. Eighty-four (29%) women were eligible for emergency contraception. Of these, 48% indicated a willingness to take emergency contraception if offered. Half of the women eligible for emergency contraception expressed ambivalent attitudes about pregnancy. Women who had taken emergency contraception in the past were more likely to say they would accept it (45%) than women who had never used it (25%, p = .05). The strongest predictor of willingness to take emergency contraception was not having a misperception about its safety, efficacy, or mechanism of action (RR = 1.9, 95% CI 1.2-3.0). Seventy-one percent of all women indicated that they would accept an advance supply of emergency contraception upon release from jail. Emergency contraception counseling and provision should be offered to newly arrested women as a key reproductive and public health intervention for a traditionally marginalized, high-risk population. PMID- 20012704 TI - Commentary re: laparoscopic versus open gastric bypass. PMID- 20012703 TI - Update in new medications for primary care. PMID- 20012705 TI - Severe peritonitis due to Streptococcus viridans following adjustable gastric banding. AB - Late postoperative intra-abdominal infections after laparoscopic gastric banding are extremely rare and may or may not be associated with the device following uncomplicated adjustable gastric banding procedures. The spectrum of pathogens associated with intra-abdominal infections is diverse and depends on the origin of the infection. Streptococci is a significant cause of peritoneal dialysis peritonitis (6% to 16% of the cases), and S. viridans is reported to account for up to 93% of streptococci infections. However, peritonitis due to S. viridans in patients following adjustable gastric banding is very rare. We herein report a non-uremic case of a 38-year-old male patient with severe peritonitis due to S. viridans 5 years after an uneventful primary gastric banding procedure. PMID- 20012706 TI - Criteria for assessing esophageal motility in laparoscopic adjustable gastric band patients: the importance of the lower esophageal contractile segment. AB - BACKGROUND: Esophageal function appears critical in laparoscopic adjustable gastric band (LAGB) patients; however, conventional motility assessments have not proven to be clinically useful. Recent combined video fluoroscopic and high resolution manometric studies have identified important components of esophageal function in LAGB patients. METHODS: Successful and symptomatic LAGB patients, with normal or mildly impaired esophageal peristalsis, underwent a standardized, water swallow, high-resolution manometry protocol designed specifically to assess the lower esophageal contractile segment (LECS), in combination with conventional measures of esophageal motility. Differences in response to changes in LAGB volume were assessed. RESULTS: There were 101 symptomatic and 29 successful patients. More symptomatic patients had a mild impairment in esophageal motility (39.6% vs. 3.4%, p < 0.005). Successful patients demonstrated an intact LECS during normal swallows more frequently than symptomatic patients (95% vs. 43%, p < 0.005). Absolute intraluminal pressures were not different between the groups. Removing all fluid from the LAGB revealed more hypotensive swallows in the symptomatic patients (30% vs. 17%, p = 0.002), an effect not observed when the LAGB volume was increased (8% vs. 5%, p = 0.21). Receiver operator characteristic analysis determined that an intact LECS in 70% of normal swallows defined normal motility in LAGB patients. CONCLUSIONS: The LECS is a valuable measure of esophageal function in LAGB patients and complements conventional manometric criteria. Symptomatic patients have less normal swallows; however, these also frequently demonstrate a deficient LECS. Further information can be elucidated by performing swallows at differing LAGB volumes. High-resolution manometry, using these adapted criteria, is now a useful in the investigation in symptomatic LAGB patients. PMID- 20012707 TI - Biorefinery: An efficient way to sustainable development of chemical industry-a special issue for international conference on biorefinery (ICB 07) and the 5th international conference on separation science and technology (ICSST2007). PMID- 20012708 TI - A perfect storm in the emergency department. AB - BACKGROUND: Serotonin syndrome is an iatrogenic disorder that results from serotonergic overactivity. Severe myoclonus and increased restlessness are hallmarks of the disorder. METHODS: We report a case of serotonin syndrome related to poor drug elimination. RESULTS: A 74-year-old white female with moderate to severe Alzheimer's dementia was brought to the emergency department for increasingly aggressive behavior. Upon admission, her risperidone dose was increased and citalopram continued. Several days later, she developed diffuse limb and facial myoclonus, spontaneous clonus, diaphoresis, and fever. Her symptoms completely resolved after discontinuation of these medications. CONCLUSION: Serotonin syndrome should be considered in the differential when elderly patients present with severe myoclonus. If unrecognized, this syndrome can lead to more severe manifestations including rhabdomyolysis, renal failure, and coma. PMID- 20012709 TI - The effect of intraventricular administration of nicardipine on mean cerebral blood flow velocity measured by transcranial Doppler in the treatment of vasospasm following aneurysmal subarachnoid hemorrhage. AB - BACKGROUND: Cerebral arterial vasospasm and delayed ischemic neurological deficits are significant contributors to morbidity and mortality following aneurysmal subarachnoid hemorrhage. Additional treatment modalities are needed. Intraventricular nicardipine has been suggested as a potential therapy for the treatment of cerebral vasospasm. It is an appealing option for multiple reasons: many of these patients already have ventricular drains in place, it can be safely administered at the bedside, and can be used in patients for whom conventional therapies are either not effective or not tolerated. METHODS: Retrospective case series of all patients who received intraventricular nicardipine for the treatment of cerebral vasospasm from January 2006 to June 2007 at a university tertiary care hospital. RESULTS: Sixty-four patients received intraventricular nicardipine during the study period. Forty-two patients met inclusion criteria. Intraventricular nicardipine administration was associated with a reduction of the mean cerebral blood flow velocity of 26.3 cm/s in the middle cerebral artery and 7.4 cm/s in the anterior cerebral artery. This reduction was maintained over 24 h with continued administration. CONCLUSIONS: Intraventricular nicardipine was associated with a significant and sustained reduction in mean cerebral blood flow velocity as measured by transcranial Doppler when used in the treatment of suspected cerebral vasospasm following aneurysmal subarachnoid hemorrhage. We do not find significant safety concerns related to elevations of intracranial pressure or ventricular catheter related infections. Further prospective studies are warranted to better determine the efficacy and safety of this therapy. PMID- 20012710 TI - Oligovalent Fab display on M13 phage improved by directed evolution. AB - Efficient display of antibody on filamentous phage M13 coat is crucial for successful biopanning selections. We applied a directed evolution strategy to improve the oligovalent display of a poorly behaving Fab fragment fused to phage gene-3 for minor coat protein (g3p). The Fab displaying clones were enriched from a randomly mutated Fab gene library with polyclonal anti-mouse IgG antibodies. Contribution of each mutation to the improved phenotype of one selected mutant was studied. It was found out that two point mutations had significant contribution to the display efficiency of Fab clones superinfected with hyperphage. The most dramatic effect was connected to a start codon mutation, from AUG to GUG, of the PelB signal sequence preceding the heavy chain. The clone carrying this mutation, FabM(GUG), displayed Fab 19-fold better and yielded twofold higher phage titers than the original Fab. PMID- 20012712 TI - Genetic variability and linkage disequilibrium patterns in the bovine DNAJA1 gene. AB - Correlation between expression level of the bovine DNAJA1 gene and meat tenderness was recently found in Charolais longissimus thoracis muscle samples, suggesting that this gene could play an important role in meat tenderness. Here, we report the validation of polymorphisms within the bovine DNAJA1 gene, and the haplotype variability and extent of linkage disequilibrium in the three main French beef breeds (Blonde d'Aquitaine, Charolais, Limousin). Genotyping 18 putative SNPs revealed that 16 SNPs were polymorphic within the breeds tested. Two SNPs were removed from further analyses as one SNP had a low genotyping call rate, while the other SNP was not in Hardy-Weinberg equilibrium. The degree of heterozygosity observed for the remaining 14 SNPs varied between breeds, with Charolais being the breed with the highest genetic variation and Blonde d'Aquitaine the lowest. Linkage disequilibrium and haplotype structure of DNAJA1 were different between breeds. Eighteen different haplotypes, including three shared by all breeds, were discovered, and two to three tag SNPs (depending on the breed) are sufficient to capture all the genetic variability seen in these haplotypes. The results of this study will facilitate the design of optimal future association studies evaluating the role of the DNAJA1 gene in meat tenderness. PMID- 20012711 TI - Mutational dynamics of microsatellites. AB - Microsatellites are a ubiquitous class of simple repetitive DNA sequences, which are widespread in both eukaryotic and prokaryotic genomes. The use of microsatellites as polymorphic DNA markers has considerably increased both in the number of studies and in the number of organisms, primarily for genetic mapping, studying genomic instability in cancer, population genetics, forensics, conservation biology, molecular anthropology and in the studies of human evolutionary history. Although simple sequence repeats have been extensively used in studies encompassing varied areas of genetics, the mutation dynamics of these genome regions is still not well understood. The present review focuses on the mutational dynamics of microsatellite DNA with special reference to mutational mechanisms and their role in microsatellite evolution. PMID- 20012713 TI - Long-term fate of allogeneic neural stem cells following transplantation into injured spinal cord. AB - To characterize the fate of allogeneic neural stem cells (NSCs) following transplantation into injured spinal cord, green fluorescent protein (GFP)-NSCs isolated from GFP transgenic Sprague-Dawley rat embryos were transplanted into contused spinal cords of Wistar rats. The GFP-NSCs survived for at least 6 months in injured spinal cord; most of them differentiated rapidly into astrocytes, and a few were able to undergo proliferation. After transplantation, the GFP-NSCs remained in the transplantation site at the early stage, and then migrated along white-matter, and gathered around the injured cavity. At 6 months post transplantation, CD8 T-lymphocytes infiltrated the spinal cord, and mixed lymphocyte culture from host and donor showed that lymphocytes from the host spleen were primed by allogeneic GFP-NSCs. At 12 months post-transplantation, most GFP cells in the spinal cord lost their morphology and disintegrated. The Basso, Beattie and Bresnahan score and footprint analysis indicated that the improvement of locomotor function in transplanted rats appeared only at the early stage, and was not seen even at 6 months after transplantation All these results suggest that the allogeneic NSCs, after transplantation into injured spinal cord, activate the host immune system. Therefore, if immunosuppressive agents are not used, the grafted allogeneic NSCs, although they can survive for a long time, are subjected to host immune rejection, and the effect of NSCs on functional recovery is limited. PMID- 20012714 TI - Presence of a ROCK inhibitor in extracellular matrix supports more undifferentiated growth of feeder-free human embryonic and induced pluripotent stem cells upon passaging. AB - Optimization and development of better defined culture methods for human embryonic and induced pluripotent stem cells (hESCs and hiPSCs) will provide an invaluable contribution to the field of regenerative medicine. However, one problem is the vulnerability of hESCs and hiPSCs to apoptosis that causes a low plating efficiency upon passaging. Herein, we have developed a novel hESCs and hiPSCs culture technique that uses ROCK inhibitor (ROCKi) Y-27632 (10 microM) in Matrigel-coated dishes in both serum- and feeder-free culture conditions. This increases plating efficiency during enzymatic and mechanical passaging as compared to its presence solely in culture medium. Under these conditions, hESCs (three lines) and hiPSCs (two lines) retain their typical morphology, a stable karyotype, express pluripotency markers and have the potential to differentiate into derivatives of all three germ layers after long-term culture. Real-time RT PCR analysis of stemness-related integrins (alphaV, alpha6, and beta1) has demonstrated that their expression increases in the presence of ROCKi. Similar plating efficiencies have been obtained in both hESCs and hiPSCs with a lower concentration of Y-27632 (800 nM) and another ROCKi (HA-1077/Fasudil), thus ruling out the non-specific effects of Y-27632. These results show that addition of ROCKi in the extracellular matrix can increase the plating efficiency of hESCs and hiPSCs during passaging of clusters. This is due not only to an anti apoptotic effect, but also to an increase in the ECM-cells interaction. Therefore, we believe this method will be useful for both current and future applications of these pluripotent stem cells. PMID- 20012715 TI - Neuroimaging of abusive head trauma. AB - The diagnostic process for evaluating suspected abusive head trauma in infants and children has evolved with technological advances in neuroimaging. Since Caffey first described a series of children with chronic subdural hematomas and multiple long bone fractures, radiologists have played an important role, along with pediatricians and pathologists, in evaluating abused children. Neuroimaging modalities include ultrasound, CT scans, and MRI technology. Each has distinct clinical applications, as well as practical uses in the clinical diagnostic process of AHT. Importantly, neuroimaging assists in the process of differential diagnosis of other conditions which may mimic AHT. Collaboration between neuroradiologists, clinicians, and pathologists remains critical to making the appropriate diagnosis. Careful history, physical examination, and investigation by legal authorities form the components that result in accurate assessment of any case. This paper reviews pertinent neuroimaging modalities currently utilized in the diagnosis of AHT, describing clinical indications and a collaborative approach to this process. PMID- 20012716 TI - Backbone assignments of the 26 kDa neuron-specific ubiquitin carboxyl-terminal hydrolase L1 (UCH-L1). AB - UCH-L1 is a member of the family of ubiquitin C-terminal hydrolases whose primary role is to hydrolyze small C-terminal adducts of ubiquitin to generate free ubiquitin monomers. Expression of UCH-L1 is highly specific to neurons and point mutations in this enzyme are associated with a hereditary form of Parkinson's disease. Herein, we present the NMR backbone assignments of human UCH-L1, thus enabling future solution-state NMR spectroscopic studies on the structure and function of this important protein. PMID- 20012718 TI - Letter to the editor: Association of osteonecrosis and failure of hip resurfacing arthroplasty. PMID- 20012719 TI - Lessons learned with extended-release epidural morphine after total hip arthroplasty. AB - An extended-release epidural morphine (EREM) has been introduced to improve postoperative pain management. Studies have shown the effectiveness of this agent in providing better pain control and patient satisfaction for patients undergoing total joint arthroplasty. We evaluated postoperative pain relief by comparing average daily pain scores and opioid use with those of the control group. Safety was measured by comparing the occurrence of postoperative complications, nausea and vomiting, pruritus, and respiratory depression between the two groups. Between February 2006 and March 2008, we selected 203 patients to receive EREM for THA. These patients were matched in a 2:1 ratio with patients undergoing THA and receiving spinal anesthesia. We retrospectively reviewed all major and minor postoperative complications from a prospective database. Patients receiving EREM had lower pain scores than patients not receiving EREM on Postoperative Day 1 (POD 1) but not POD 2, or POD 3. Patients receiving EREM experienced a slightly higher incidence of pulmonary embolism and supraventricular tachycardia. Patients receiving EREM also experienced more nausea and vomiting and pruritus. We found EREM provided better pain relief on POD 1 at the expense of a slightly higher incidence of side effects compared with spinal anesthesia alone. LEVEL OF EVIDENCE: Level III, therapeutic study. See the Guidelines for Authors for a complete description of levels of evidence. PMID- 20012720 TI - Hip resurfacing data from national joint registries: what do they tell us? What do they not tell us? AB - Current-generation metal-on-metal hip resurfacing implants (SRAs) have been in widespread global use since the 1990s, and in the United States, specific implants have recently been approved for clinical use. Many recent publications describe short-term survivorship achieved by either implant-designing surgeons or high-volume centers. National joint replacement registries (NJRRs) on the other hand report survivorship achieved from the orthopaedic community at large. We therefore reviewed SRA survivorship from national registry data and compared with that reported from individual centers. Additionally, we compared SRA survivorship reported by registries and compared prognosticators for failure with those of conventional total hip arthroplasty (THA). Although resurfacing was associated with an overall increased failure rate in comparison to THA (Australian registry 5-year cumulative revision rate [CRR], 3.7% and 2.7%, respectively), there were exceptions to this. Male patients younger than 65 years with primary osteoarthritis had equivalent results with SRA and THA (Australian registry 5 year CRR, 2.5% and 2.8%, respectively). Head size over 50 mm in diameter was a predictor of surface replacement arthroplasty survivorship and only females with a head diameter of 50 mm or greater (14% of females) had a comparable survivorship to males. Diagnoses other than primary osteoarthritis bear a higher risk of early revision of SRA as compared with THA. Revision of SRA does not lead to reproducible results with rerevision rates of 11% at 5 years. Given these predictors of failure, our review of data from the NJRR suggests stringent patient selection criteria might enhance the survival rates of SRA. PMID- 20012721 TI - Antioxidant capacity and oxidative damage determination in synovial fluid of chronically damaged equine metacarpophalangeal joint. AB - In order to determine oxidative stress in equine joints with degenerative processes, we analyzed synovial fluid (SF) antioxidant capacity and the concentration of oxidative damage biomarkers in healthy and chronically damaged metacarpophalangeal joints. SF samples were collected from joints of thirty 2-5 year-old crossbreed male equine, macroscopically classified at post mortem inspection and later histologically confirmed. The antioxidant capacity was determined measuring uric acid and the concentration of sulfhydryl groups and the total radical trapping antioxidant potential (TRAP). The oxidative damage was determined by assessing malondialdehyde (MDA) and carbonyl protein concentration. TRAP was significantly higher (p < 0.05) in the group with chronic damage (CD). The sulfhydryl groups and concentration of uric acid did not show significant difference between the groups (p > 0.05). Although carbonyl concentration did not show significant difference between groups, it was slightly higher in the group with CD (p = 0.05009). Concentration of MDA did not show significant difference (p > 0.05) between groups. The observed significant increase in TRAP in the group with CD could be related to the participation of components other than protein, sulfhydryl groups, or uric acid coming from degenerating joint tissues. These findings could be helpful for a better understanding of the oxidative stress role in equine joints with chronic degenerative process. PMID- 20012722 TI - Arsenic trioxide inhibits cholangiocarcinoma cell growth and induces apoptosis. AB - Arsenic trioxide (As(2)O(3)), an ingredient in many traditional Chinese medicines, has drawn broad attention due to its therapeutic effects on a variety of cancers, including some solid tumors. However, the effects of As(2)O(3) on cholangiocarcinoma have not been reported. In the present study, we demonstrate for the first time that clinically obtainable concentrations of As(2)O(3) inhibit cell growth and induce apoptosis in human cholangiocarcinoma SK-ChA-1 cells. As(2)O(3)-induced apoptosis was partially inhibited by caspase inhibitor and accompanied by changes in the expression of Bcl-2 family proteins, decrease of mitochondrial membrane potential (MMP), release of cytochrome C from mitochondria, activation of caspase-3, caspase-9, and cleavage of poly (ADP ribose) polymerase (PARP). Thus As(2)O(3) induces apoptosis in SK-ChA-1 cells via mitochondria-mediated, caspases-dependent pathways. As(2)O(3) inhibition of Akt phosphorylation may contribute to As(2)O(3)-mediated cholangiocarcinoma cell growth inhibition and apoptosis induction. PMID- 20012723 TI - An investigation of potential sources of artifacts in SPECT-CT myocardial perfusion studies. AB - BACKGROUND: The increased use of hybrid SPECT-CT systems in myocardial perfusion imaging calls for a careful review of protocols that are employed in data acquisition and processing. Our study investigates the cases of potential false perfusion defects that may appear in cardiac images reconstructed with CT-based attenuation correction and high-resolution acquisition matrix. METHODS: Phantom experiments performed on Infinia-Hawkeye (GE Healthcare) modeling patients with normal hearts and different body sizes and shapes were reconstructed using standard ordered subsets expectation maximization (OSEM) method with 10 subsets and 2-50 iterations. The CT-based attenuation correction (AC) with and without distance-dependent resolution recovery (RR) were employed. RESULTS: Images reconstructed from scans with a standard thorax phantom did not show any artifacts. However, some images reconstructed from the data with extra water bags (modeling high and non-uniformly distributed attenuation) clearly displayed false perfusion defects. A potential for creating such artifacts was especially high in complex reconstruction cases. In most cases, the severity of defects decreased when reconstructions with more iterations were used. CONCLUSIONS: In situations with strong and non-uniform attenuation, the reconstruction methods with sophisticated data processing (large matrix size, AC + RR corrections) may require substantially more processing than is currently used and/or recommended. PMID- 20012724 TI - Antimicrobial susceptibility of pathogens isolated from more than 10,000 patients with infectious respiratory diseases: a 25-year longitudinal study. AB - The Study Group on Antimicrobial Susceptibility of Pathogens Isolated from Respiratory Infections was established in 1981 in Japan to elucidate trends in such susceptibilities in patients with infectious respiratory diseases; the Group has conducted nationwide research in collaboration with 21 medical institutions. Examination of more than 10,000 patients by 2005 allowed a summary of study findings. Streptococcus pneumoniae started to become resistant to penicillin G in the 1990s, and the isolation rate of penicillin-intermediate and penicillin resistant S. pneumoniae (PISP + PRSP) reached almost 60% in 2001. The proportion of PRSP also increased, reaching 19.4%. Thereafter, the rate of PISP + PRSP decreased somewhat to the mid-30% range. Macrolide resistance was also observed; in 2005, the prevalence of strains highly susceptible to erythromycin with MICs or= 128 microg/ml exceeded 40%. Among Staphylococcus aureus isolates, the proportion of methicillin-resistant S. aureus (MRSA) strains began to increase rapidly in 1986 and constituted around 60% of all S. aureus strains identified in 1990 and in the following years. In 1993, the prevalence of ampicillin-resistant isolates of Haemophilus influenzae had increased remarkably, presumably related to the outbreak of beta-lactamase-negative ampicillin resistant (BLNAR) H. influenzae strains, and the proportion of these strains among the isolates surpassed 30% in 2002 and thereafter. For Klebsiella pneumoniae, the antimicrobial activity of first- to fourth-generation cephems improved with each generation. The MIC distribution patterns of Pseudomonas aeruginosa shifted towards higher MICs when compared with the MICs for other pathogens. Broad patterns with no distinct peaks reflected the difficulty in treating P. aeruginosa infection. Regarding Moraxella catarrhalis, beta-lactamase producing strains already constituted a majority of the isolates in 1990, and the proportion of strains highly susceptible to ampicillin, with MICs or=5.0 microg/ml for 1 h, mRNA expression levels of CXCR1/2 and TNFalpha were suppressed in a dose-dependent manner. The gene expression level of CD14 was also downregulated by LPZ at >or=0.1 microg/ml, with expression suppressed to 50% by 10 microg/ml LPZ. However, LPZ at 0.01-5.0 microg/ml had no significant effect on the expression of TLR-4 or CD11b/CD18 mRNA. LPZ at 10 microg/ml downregulated the levels of these mRNAs to 80% and 50%, respectively. On the other hand, when the reaction period was extended to 3 h with the same conditions, all mRNA expression levels were downregulated by >or=0.01 microg/ml LPZ, in a dose-dependent manner. LPZ may suppress the biological functions of PMNs, such as chemotaxis and inflammatory chemokine production. PMID- 20012728 TI - A comparative clinical study of macrolide-sensitive and macrolide-resistant Mycoplasma pneumoniae infections in pediatric patients. AB - In recent years, the increased prevalence of macrolide-resistant Mycoplasma pneumoniae (MR-M. pneumoniae) has become a significant issue in Japan. We isolated 94 strains of M. pneumoniae, and determined the minimum inhibitory concentrations (MICs) of macrolides and other antimicrobial agents for these strains. We also performed a comparative clinical evaluation of macrolide efficacy for cases of MR-M. pneumoniae infections and cases of macrolide sensitive Mycoplasma pneumoniae infections (MS-M. pneumoniae). Of the 94 isolates of M. pneumoniae, 64 (68.1%) were classified as MS-M. pneumoniae and 30 (31.9%) as MR-M. pneumoniae strains. The clinical study included an assessment of 47 pediatric cases of MS-M. pneumoniae and 22 pediatric cases of MR-M. pneumoniae. The patient demographics, such as sex, age, the period from the onset of the infection to the first examination, laboratory findings, diagnosis, and the severity of symptoms, showed no significant difference between the two study groups. However, the efficacy of macrolide treatment was 91.5% for MS-M. pneumoniae and 22.7% for MR-M. pneumoniae, a statistically significant difference (P < 0.01). Although M. pneumoniae infection is generally considered a treatable condition, the increasing prevalence of macrolide-resistant strains of M. pneumoniae has become a significant clinical issue in pediatric patients, and it is therefore necessary to give careful consideration to the appropriate antimicrobial therapy for MR-M. pneumoniae infection. PMID- 20012729 TI - Fact-finding survey of nosocomial infection control in hospitals in Vietnam and application to training programs. AB - Nosocomial infection control is crucial for improving the quality of medical care. It is also indispensable for implementing effective control measures for severe acute respiratory syndrome (SARS) and the possible occurrence of a human influenza pandemic. The present authors, in collaboration with Vietnamese hospital staff, performed a fact-finding survey of nosocomial infection control in hospitals in northern Vietnam and compared the results with those of a survey conducted 4 years previously. Remarkable improvement was recognized in this period, although there were considerable differences between the central hospitals in Hanoi and local hospitals. In the local hospitals, basic techniques and the systems for infection control were regarded as insufficient, and it is necessary to improve these techniques and systems under the guidance of hospitals in the central area. Based on the results of the survey, programs were prepared and training courses were organized in local hospitals. Evaluation conducted after the training courses showed a high degree of satisfaction among the trainees. The results of the survey and the training courses conducted during the study period are expected to contribute to the improvement of nosocomial infection control in remote areas of Vietnam. PMID- 20012730 TI - Efficacy of treatment with carbapenems and third-generation cephalosporins for patients with febrile complicated pyelonephritis. AB - The purpose of this study was to clarify the clinical relevance of carbapenem and third-generation cephalosporin treatment for febrile complicated pyelonephritis, which often leads to urosepsis. Parenteral antimicrobial treatment with a carbapenem or third-generation cephalosporin was administered to febrile patients and the treatment was switched to oral antimicrobial agents after they became afebrile. In principle, the duration of the course of antimicrobial chemotherapy was limited to a total of 14 days. Clinically, the success rates were 97.3% in the carbapenem group and 96.0% in the third-generation cephalosporin group. For microbiological efficacy, the success rates were 89.2% in the carbapenem group and 92.0% in the third-generation cephalosporin group. There were no serious adverse events in the course of the study. The treatment regimen with a carbapenem or a third-generation cephalosporin was highly effective for patients with febrile complicated pyelonephritis and was well tolerated. Either of these regimens could become one of the standard treatments for patients with febrile complicated pyelonephritis. PMID- 20012731 TI - Nationwide survey of the development of drug-resistant pathogens in the pediatric field: drug sensitivity of Streptococcus pneumoniae in Japan. AB - We evaluated the resistance to 20 different antibacterial agents of 362 clinically isolated strains of Streptococcus pneumoniae accumulated from October 2000 to July 2001 (phase 1) and of 332 different strains accumulated from January to June 2004 (phase 2), from institutions throughout Japan that participated in the surveys carried out by the Drug-Resistant Pathogen Surveillance Group in Pediatric Infectious Disease. In phase 1, the proportions of penicillin-sensitive S. pneumoniae (PSSP), penicillin-insensitive S. pneumoniae (PISP), and penicillin resistant S. pneumoniae (PRSP) were 35.4%, 34.8%, and 29.8%, respectively, and the proportions were almost the same in phase 2: 33.1%, 37.0%, and 29.8%, respectively. Comparison of the MIC(90) values of the antibacterial agents for PRSP in phase 1 and phase 2 revealed that these values for cefditoren, cefpodoxime, cefdinir, faropenem, ceftriaxone, cefotaxime, meropenem, and vancomycin increased by twofold to fourfold during the 3 years between phase 1 and phase 2. However the MIC(90) of rokitamycin increased more than fourfold. The proportion of S. pneumoniae that were PISP + PRSP remained almost constant over the 3 years between phase 1 and phase 2. The background factors of patient age, previous administration of antibacterial agents, and attendance at a day nursery were examined; we found that in phase 1, the proportion of PISP + PRSP was significantly higher than that of PSSP in patients under 4 years old who had previously received antibacterial agents, but no significant differences were found in any of these background factors in the phase 2 survey. No significant difference was found in the proportions of penicillin-resistant bacteria according to whether or not the child had attended a day nursery. PMID- 20012732 TI - Nationwide survey of the development of drug-resistance in the pediatric field: drug sensitivity of Haemophilus influenzae in Japan. AB - We evaluated the beta-lactamase-producing ability and resistance to 20 antibacterial agents of 448 clinically isolated strains of Haemophilus influenzae accumulated from October 2000 to July 2001 (phase 1) and of 376 different strains accumulated from January to June 2004 (phase 2), from institutions that participated in a nationwide Drug-Resistant Pathogen Surveillance Group in Pediatric Infectious Disease. Between phase 1 and phase 2 the proportion of beta lactamase-negative ampicillin (ABPC)-susceptible (BLNAS) strains declined from 62.9% to 34.3%; the proportions of beta-lactamase-positive ABPC-resistant (BLPAR) strains were 8.3% and 6.4% in phases 1 and 2, but the proportion of beta lactamase-negative ABPC-resistant (BLNAR) strains increased from 28.8% in phase 1 to 59.3% in phase 2. Comparison of the MIC(90) values of the antibacterial agents for H. influenzae in phase 1 and phase 2 showed that cefcapene, cefpodoxime, ceftriaxone, panipenem, and clarithromycin kept the same level, while cefdinir, faropenem, and rokitamycin showed 2-fold to 8-fold decreases. With the exception of the above antibiotics, all of the other antibacterial agents tested showed 2 fold to 4-fold increases. The MIC(90) values of the beta-lactam drugs for BLNAR were 2-fold to 32-fold higher than the values for BLNAS. The proportion of BLNAR H. influenzae strains rose dramatically over the 3 years between phases 1 and 2. In relation to age, prior administration of antibacterial agents, and attendance at a day nursery as background factors, no significant differences between BLNAS and BLNAR were detected in phase 1. In the phase 2 survey, the proportion of BLNAR strains showed significant differences between children under 3 years and those aged 3 years or more, and there were also significant differences according to whether antibacterial agents, especially beta-lactams, had been administered previously. No significant difference was found in resistant bacteria according to whether or not a child had attended a day nursery. PMID- 20012733 TI - Failure to detect Mycoplasma genitalium in the pharynges of female sex workers in Japan. AB - To determine the prevalence of genital mycoplasmas and ureaplasmas in the pharynges of Japanese female sex workers practicing fellatio on their clients, vaginal swabs and throat washings were collected from 403 female sex workers attending a clinic in Kyoto, Japan, for regular screening of gonococcal and chlamydial infections. Neisseria gonorrhoeae, Chlamydia trachomatis, Mycoplasma genitalium, Mycoplasma hominis, Ureaplasma parvum, and Ureaplasma urealyticum in vaginal and throat specimens were tested by nucleic acid amplification tests. The prevalence of N. gonorrhoeae, C. trachomatis, M. genitalium, M. hominis, U. parvum, and U. urealyticum in the genitals was 1.7%, 7.2%, 1.7%, 19.6%, 40.4%, and 10.2%, respectively, whereas their prevalence in the pharynges was 4.0%, 2.0%, 0%, 1.2%, 0.2%, and 0.7%, respectively. Gonococcal infection in the pharynx was significantly associated with gonococcal infection in the genitals. Chlamydial infection in the pharynx was also significantly associated with chlamydial infection in the genitals. M. hominis, U. parvum, and U. urealyticum were all detected in vaginal swabs and in throat washings; however, M. genitalium was detected in vaginal swabs but not in throat washings. For each of these genital mycoplasmas and ureaplasmas, a positive test result in the pharynx was not significantly associated with a positive result in the genitals. M. hominis, U. parvum, and U. urealyticum were detected in throat washings, but M. genitalium was not. These findings do not necessarily rule out the transmission of M. genitalium from the pharynx to the urethra by orogenital sex. PMID- 20012734 TI - Cat scratch disease presenting with a retroperitoneal abscess in a patient without animal contacts. AB - Cat scratch disease (CSD) is usually diagnosed in patients presenting with regional lymphadenopathy and pyrexia that follow contacts with animals. We describe here a young adult male patient who presented with marked pyrexia and a retroperitoneal abscess without relevant medical histories, illustrating that CSD can be a diagnostic challenge on selected occasions. PMID- 20012735 TI - Linezolid-resistant methicillin-resistant Staphylococcus aureus isolated after long-term, repeated use of linezolid. AB - We report a 54-year-old male patient with an infection caused by linezolid resistant methicillin-resistant Staphylococcus aureus (MRSA), isolated after long term, repeated use of linezolid. Five MRSA strains isolated from our patient were preserved and submitted to bacteriological analysis. All five of these strains were found to have identical genotypes by pulsed-field gel electrophoresis. Two strains isolated in the early hospital period were sensitive to linezolid, while three isolated in the late hospital period were resistant. These three strains that had acquired resistance to linezolid were found to have a G2576T point mutation in the 23SrRNA domain V. Linezolid-resistant MRSA is rare, but may occur with the long-term, repeated administration of linezolid. PMID- 20012736 TI - Pharmacokinetics and pharmacodynamics of once-daily arbekacin during continuous venovenous hemodiafiltration in critically ill patients. AB - This study examined the pharmacokinetics of arbekacin during continuous venovenous hemodiafiltration (CVVHDF) and assessed the pharmacodynamics to consider arbekacin dosage adaptation in CVVHDF. Arbekacin was administered by 0.5 h infusion once daily, using a polymethyl methacrylate membrane hemofilter, to three critically ill patients undergoing CVVHDF; the flow rates were 0.8 l/h for the filtrate and 0.6 l/h for the dialysate. The drug concentrations in plasma and in the filtrate-dialysate were determined using a fluorescence polarization immunoassay and analyzed pharmacokinetically. The average sieving coefficient of arbekacin was 0.739 and the average drug clearance by CVVHDF was 1.03 l/h. A pharmacokinetic model with three compartments (1, central; 2, peripheral; 3, filtrate-dialysate side hemofilter) accurately reflected the concentration-time data for both plasma and filtrate-dialysate. The pharmacokinetic model assessed the pharmacodynamic profile of arbekacin once-daily regimens (0.5-h infusions) at filtrate-dialysate flow rates of 1.4 and 2.8 l/h, and demonstrated that only the 150-mg and 200-mg regimens achieved an effective target range for C(max) (9-20 microg/ml), suggesting that empirical dosages lower than the usual 150-200 mg should be avoided in patients undergoing CVVHDF. The minimum regimens needed to achieve an effective pharmacodynamic target for the free C(max)/MIC ratio (>8) were 75 mg for an MIC of 0.5 microg/ml, 200 mg for an MIC of 2 microg/ml, and 400 mg for an MIC of 4 microg/ml. These results will help us to better understand the pharmacokinetics of arbekacin during CVVHDF, while also helping in the selection of the appropriate arbekacin regimens, based on a pharmacodynamic assessment, for patients receiving this renal replacement therapy. PMID- 20012737 TI - Comparative analysis of Micrococcus luteus isolates from blood cultures of patients with pulmonary hypertension receiving epoprostenol continuous infusion. AB - During the period 2002-2008, at the National Cardiovascular Center, Osaka, 28 Micrococcus luteus isolates and one Kocuria spp. isolate were obtained from blood cultures of pulmonary hypertension (PH) patients who were receiving continuous infusion therapy with epoprostenol. Pulsed-field gel electrophoresis patterns of the isolates were unrelated, suggesting that the infections had multiple origins. The preparation of epoprostenol solution by patients themselves was thought to be a risk factor. PMID- 20012738 TI - Procalcitonin and severity of community-acquired pneumonia. AB - The purpose of this study was to clarify the relationship between procalcitonin and the severity and prognosis of community-acquired pneumonia. The subjects were 162 patients with community-acquired pneumonia (disease severity, mild, 39 patients; moderate, 81 patients; severe, 37 patients; and super severe, 5 patients) in whom we examined the serum procalcitonin concentration at the start of treatment; we determined the relationship of procalcitonin status with disease severity and prognosis. The results showed that procalcitonin was positive in 12.8% of the patients with mild disease, 27.1% of the patients with moderate disease, 59.5% of the patients with severe disease, and 80.0% of the patients with super severe disease. The mortality of procalcitonin-positive patients was 37.7%, whereas that of the procalcitonin-negative patients was 12.8%. Based on the above findings, it is concluded that the more severe the community-acquired pneumonia, the higher is the positivity rate for procalcitonin, and the prognosis in procalcitonin-positive patients is worse than that in procalcitonin-negative patients. PMID- 20012739 TI - Mesenchymal stromal cells derived from human umbilical cord tissues: primitive cells with potential for clinical and tissue engineering applications. AB - Mesenchymal stem or stromal cells (MSCs) have a high potential for cell-based therapies as well as for tissue engineering applications. Since Friedenstein first isolated stem or precursor cells from the human bone marrow (BM) stroma that were capable of osteogenesis, BM is currently the most common source for MSCs. However, BM presents several disadvantages, namely low frequency of MSCs, high donor-dependent variations in quality, and painful invasive intervention. Thus, tremendous research efforts have been observed during recent years to find alternative sources for MSCs.In this context, the human umbilical cord (UC) has gained more and more attention. Since the UC is discarded after birth, the cells are easily accessible without ethical concerns. This postnatal organ was found to be rich in primitive stromal cells showing typical characteristics of bone-marrow MSCs (BMSCs), e.g., they grow as plastic-adherent cells with a fibroblastic morphology, express a set of typical surface markers, and can be directly differentiated at least along mesodermal lineages. Compared to BM, the UC tissue bears a higher frequency of stromal cells with a higher in vitro expansion potential. Furthermore, immune-privileged and immune-modulatory properties are reported for UC-derived cells, which open highly interesting perspectives for clinical applications. PMID- 20012740 TI - A complex genetic network involving a broad-spectrum locus and strain-specific loci controls resistance to different pathotypes of Aphanomyces euteiches in Medicago truncatula. AB - A higher understanding of genetic and genomic bases of partial resistance in plants and their diversity regarding pathogen variability is required for a more durable management of resistance genetic factors in sustainable cropping systems. In this study, we investigated the diversity of genetic factors involved in partial resistance to Aphanomyces euteiches, a very damaging pathogen on pea and alfalfa, in Medicago truncatula. A mapping population of 178 recombinant inbred lines, from the cross F83005.5 (susceptible) and DZA045.5 (resistant), was used to identify quantitative trait loci for resistance to four A. euteiches reference strains belonging to the four main pathotypes currently known on pea and alfalfa. A major broad-spectrum genomic region, previously named AER1, was localized to a reduced 440 kb interval on chromosome 3 and was involved in complete or partial resistance, depending on the A. euteiches strain. We also identified 21 additive and/or epistatic genomic regions specific to one or two strains, several of them being anchored to the M. truncatula physical map. These results show that, in M. truncatula, a complex network of genetic loci controls partial resistance to different pea and alfalfa pathotypes of A. euteiches, suggesting a diversity of molecular mechanisms underlying partial resistance. PMID- 20012741 TI - Influence of salt ions on binding to molecularly imprinted polymers. AB - Salt ions were found to have an influence on template binding to two model molecularly imprinted polymers (MIPs), targeted to penicillin G and propranolol, respectively, in water-acetonitrile mixtures. Water was detrimental to rebinding of penicillin G whereas propranolol bound in the entire water-acetonitrile range tested. In 100% aqueous solution, 3-M salt solutions augmented the binding of both templates. The effects followed the Hofmeister series with kosmotropic ions promoting the largest increase. Binding was mainly of a non-specific nature under these conditions. In acetonitrile containing low amounts of water, the specific binding to the MIPs increased with the addition of salts. Binding of penicillin G followed the Hofmeister series while an ion-exchange mechanism was observed for propranolol. The results suggest that hydration of kosmotropic ions reduces the water activity in water-poor media providing a stabilizing effect on water sensitive MIP-template interactions. The effects were utilized to develop a procedure for molecularly imprinted solid-phase extraction (MISPE) of penicillin G from milk with a recovery of 87%. PMID- 20012742 TI - Synchrotron FTIR microspectroscopy of the yeast Saccharomyces cerevisiae after exposure to plasma-deposited nanosilver-containing coating. AB - The present work was focused on elucidating changes in the model yeast Saccharomyces cerevisiae (cell composition, ultrastructure) after exposure to antimicrobial plasma-mediated nanocomposite films. In order to achieve this, a nanosilver-containing coating was deposited onto stainless steel using radiofrequency HMDSO plasma deposition, combined with simultaneous silver sputtering. X-ray photoelectron spectroscopy (XPS) confirmed the presence of silver nanoparticles embedded in an organosilicon matrix. In addition, scanning electron microscopy (SEM) demonstrated the nanoparticle-based morphology of the deposited layer. The antifungal properties towards S. cerevisiae were established, since a 1.4 log reduction in viable counts was observed after a 24-h adhesion compared to control conditions with the matrix alone. Differences in cell composition after exposure to the nanosilver was assessed for the protein region using, for the first time, synchrotron Fourier-transform infrared (FTIR) microspectroscopy of single S. cerevisiae cells, through in situ mapping with sub cellular spatial resolution. IR spectrum of yeast cells recovered after a 24-h adhesion to the nanosilver-containing coating revealed a significant downshift (20 cm(-1)) of the amide I peak at 1655 cm(-1), compared to freshly harvested cells. This lower band position, corresponding to a loss in alpha-helix structures, is indicative of the disordered secondary structures of proteins, due to the transition between active and inactive conformations under nanosilver induced stress conditions. No significant effect on the nucleic acid region was detected. The inhibitory action of silver was targeted against both cell wall and intracellular proteins such as enzymes. Transmission electron microscopy (TEM) observations of the yeast ultrastructure confirmed serious morphological and structural damages. A homogeneous protein-binding distribution of nanosilver all over the cell was assumed, since the presence of electron-dense silver clusters was detected not only on the cell surface but also within the cell. For control experiments with the organosilicon matrix alone, no antimicrobial effect was observed, which was consistent with synchrotron FTIR results and TEM observations. PMID- 20012743 TI - Toxicity of octahydro-1,3,5,7-tetranitro-1,3,5,7-tetrazocine (HMX) in three vertebrate species. AB - The explosive, octahydro-1,3,5,7-tetranitro-1,3,5,7-tetrazocine or high-melting explosive (HMX), has been found in soils in areas used for testing and training by the military. Many of these areas contain habitat for valued wildlife species. In an effort to better understand the environmental consequences from exposure, a reptilian (western fence lizard [Sceloporus occidentalis]), an amphibian (red backed salamander [Plethodon cinereus]), and a mammalian species (rabbit [Oryctolagus cuniculus]) were exposed to HMX under controlled laboratory conditions. Lizards and rabbits were exposed to HMX by way of corn oil through gavage, and salamanders were exposed to HMX in soil. Two deaths occurred from acute oral exposures to lizards to 5000 mg HMX/kg BW. Histological and gross pathologic assessment suggested gut impaction as a possible cause of death. Salamanders exposed to concentrations of HMX in soil < or = 1970 mg HMX/kg soil for 10 days did not show adverse effects. Rabbits, however, showed neurologic effects manifested as hyperkinetic events with convulsions at > 24 h after oral exposures. An LD(50) for rabbits was calculated as 93 mg/kg (95% confidence interval 76-117). A subacute 14-day testing regime found a lowest observed effect level of 10 mg/kg-d and a no observed adverse effect level of 5 mg/kg-d based on hyperkinesia and seizure incidence, although changes suggesting functional hepatic alterations were also found. These data suggest that physiologic differences between species, particularly in gastrointestinal structure and function, can affect the absorption of HMX and hence lead to marked differences in toxicity from exposure to the same compound. PMID- 20012744 TI - Adsorption of phorate, an organophosphorus pesticide, on vertisol. AB - Adsorption of phorate, an organophosphorus pesticide, on a vertisol soil was studied. The resulting data were well described by Freundlich and Langmuir adsorption isotherms. Adsorption was fast and the equilibrium was established within 8 h, which is comparatively less than reported previously. The mechanism of interaction between phorate and clay and humic acid extracted from the same soil was studied by Fourier-transform infrared (FTIR) spectroscopy. FTIR results suggested the formation of hydrogen bonds between carboxylic acid groups present in humic acid and appropriate electrophilic hydrogen atoms present in phorate. Also there is an indication of involvement of -P-O- group of phorate in the interaction with humic acid. However, the binding of phorate with clay minerals involves van der Waal forces of attraction. PMID- 20012746 TI - Anemia response and safety to epoetin-beta treatment in patients with neoadjuvant therapy prior to primary digestive tract tumor surgery. AB - PURPOSE: Anemia is common during anticancer treatment. This study aimed to evaluate the response and safety of treatment with epoetin-beta (EB) in patients with neoadjuvant therapy prior to primary digestive tract tumor surgery. PATIENTS AND METHODS: In this open-label, single-arm study, patients (n = 22) with hemoglobin (Hb) levels below 11 g/dl who received epoetin-beta 450 IU/kg (30,000 IU) weekly until the hemoglobin level reached 12 g/dl. RESULTS: After treatment with EB, a mean absolute increment of 2.6 g/dl was attained. The mean hemoglobin values during the study were pretreatment 10.1 g/dl, half-way through treatment 12.3 g/dl, 4 weeks after concomitant radiochemotherapy 12.7 g/dl, the week prior to surgery 12.5 g/dl, and after surgery 10.9 g/dl. No patient required transfusion before or after surgery. The probability or risk of postoperative complications was 27.3%, and included one rectovaginal fistula, one parastomal hernia, one case of ileus and two surgical wound infections. In this series, downstaging was observed in 81.8% of patients, and downsizing in 90.9%. Most interestingly, histopathological complete response rate was achieved by 18.2%. CONCLUSIONS: Epoetin-beta (EB) treatment in our series of patients with digestive malignancies subjected to neoadjuvant radiochemotherapy proved effective and safe, avoiding the need for transfusion during surgery. PMID- 20012745 TI - Nonunions around the knee joint. AB - Nonunions of the distal femur are relatively rare but can occur with or without previous surgery. Many surgical methods have been described for the treatment of these nonunions including external fixation, intramedullary nailing, arthroplasty, and fixed angle plating. While arthroplasty may have a limited role in the elderly patient with osteoporosis and severe communiution, most nonunions can be reliably treated with revision fixation using a fixed angled device, lag screws, and bone grafting as needed. Fixed-angle plating also has the advantage of stabilising a small distal segment where intramedullary nailing has proven to be less effective. Any surgical intervention must include restoration of the normal mechanical axis of the distal femur for a satisfactory clinical outcome. Proximal tibia nonunions are a similarly rare clinical entity due to the excellent blood supply in the metaphysis. Extra-articular proximal tibial nonunions are often due to malreduction during the index surgery resulting in poor bone contact and biomechanical stability. Soft tissue forces and improper use of intramedullary implants often predispose these fractures to malalignment. Revision surgery consisting of deformity correction and at least a lateral tension band plate with lag screws has led to predictable union and good clinical results. Tibial plateau nonunions are exceedingly uncommon but are difficult to treat because of small, often devascularised fragments, post-surgical scarring, and osteopenia. Revision surgery with meticulous soft tissue handling to restore the articular surface and mechanical axis with repair of meniscal pathology gives the patient the best chance of recovering knee function. PMID- 20012747 TI - Outcome of preemptive penile rehabilitation before bilateral cavernosal nerve injury in rats. AB - PURPOSE: In this study, prophylactic penile rehabilitation (PR) with sildenafil before and after a cavernosal nerve (CN) injury was analyzed in an animal model. MATERIALS AND METHODS: Thirty-six animals were divided into six groups as follows: (1) those with no CN injury (i.e., sham), (2) those with a bilateral CN injury (i.e., control), (3, 4) those with a bilateral CN injury treated with 10 20 mg/kg of sildenafil subcutaneously (SC) on a daily basis commencing 1 month prior to and after nerve injuries, respectively, (5, 6) those with a bilateral CN injury treated daily with 10-20 mg/kg of sildenafil SC after the nerve injuries, respectively. Mean arterial pressure (MAP) and intracavernosal pressure (ICP) were measured in response to CN stimulation to assess erectile function (EF). Neuronal nitric oxide synthase (nNOS) immunohistochemical analysis and real-time polymerase chain reaction (RT-PCR) were performed. RESULTS: The maximal ICP/MAP ratio was 60 +/- 18% in the sham, 22 +/- 5% in the control, 28 +/- 9% in Group III, 45 +/- 16% in Group IV, 45 +/- 17% in Group V, and 49 +/- 21% in Group VI. Although EF was improved with sildenafil treatment in a dose-dependent fashion, no statistically significant difference was observed between the preemptive and standard rehabilitation groups. Again, nNOS immunoreactivity and RT-PCR results showed the beneficial effect of sildenafil, but the study did not support the efficacy of preemptive rehabilitation when compared to the standard rehabilitation group. CONCLUSIONS: Although, a dose-response relationship was observed for PR treatment with sildenafil; i.e., outcomes improved at higher doses of sildenafil for PR, preemptive PR should not be pursued as an alternative rehabilitation modality. PMID- 20012748 TI - Anti-inflammatory effect of oxytocin in rat myocardial infarction. AB - While an increasing amount of evidence demonstrates the homeostatic functions of the cardiac oxytocin (OT) system, less is known about the role of this hormone in the injured heart. The current study examined the effect of OT infusion on cell apoptosis, expression of proliferating cell nuclear antigen (PCNA) and inflammation in the acute and subacute phases of myocardial infarction (MI). Prior MI male Sprague-Dawley rats were infused subcutaneously with OT 25 or 125 ng/(kg h) for 3 or 7 days. Saline-treated MI and sham-operated rats served as controls. Echocardiography and analysis of cardiac sections were used to disclose OT actions. Left ventricular fractional shortening, estimated to be 46.0 +/- 1.8% in sham controls, declined to 21.1 +/- 3.3% in vehicle-treated MI rats and was improved to 34.2 +/- 2.1 and to 30.9 +/- 2.5% after treatment with OT 25 and 125 ng/(kg h), respectively. OT infusion resulted in: (1) increase of cells expressing PCNA in the infarct zone, diminished cell apoptosis and fibrotic deposits in the remote myocardium; (2) suppression of inflammation by reduction of neutrophils, macrophages and T lymphocytes; (3) depression of the expression of proinflammatory cytokines tumor necrosis factor-alpha and interleukin-6 with promotion of transforming growth factor-beta. OT treatment reduced expression of atrial and brain natriuretic peptides in the infarcted ventricle, as well as the concentration of both peptides in the circulation. These results indicate that continuous OT delivery reduces inflammation and apoptosis in infarcted and remote myocardium, thus improving function in the injured heart. PMID- 20012749 TI - The skin pathergy test: innately useful? AB - Pathergy is the term used to describe hyper-reactivity of the skin that occurs in response to minimal trauma. A positive skin pathergy test (SPT), characterised by erythematous induration at the site of the needle stick with a small pustule containing sterile pus at its centre, is among the criteria required for a diagnosis of Behcet's disease (BD) and in certain population has been shown to be highly specific for this condition. Problems with standardising the induction manoeuvre for the SPT as well as the method of assessment of the response have limited the usefulness of the SPT in the clinical setting. Extensive investigation into histopathological and immunological aspects of pathergy has led to a number of hypotheses relating to the aetiology of the skin pathergy reaction and the disease itself, but the cause is considered to be unknown. Pathergy lesions, the development of new skin lesions or the aggravation of existing ones following trivial trauma, are also reported in pyoderma gangrenosum and has been noted in other neutrophilic dermatoses such as Sweet's syndrome. The response of such patient groups to the systematic application of the SPT has not been described. We propose that a new way of considering the pathergy reaction is to see it as an aberration of the skin's innate reactivity from a homeostatic reactive mode closely coupled to tissue healing to an abnormal destructive/inflammatory mode. Our understanding of BD and other similar conditions would profit by more detailed mechanistic knowledge of skin homeostasis to minimal trauma in both health and disease through a more structured and systematic use of the SPT. PMID- 20012750 TI - Leukemia incidence among people exposed to chronic radiation from the contaminated Techa River, 1953-2005. AB - Beginning in 1950, people living on the banks of the Techa River received chronic low-dose-rate internal and external radiation exposures as a result of releases from the Mayak nuclear weapons plutonium production facility in the Southern Urals region of the Russian Federation. The Techa River cohort includes about 30,000 people who resided in riverside villages sometime between 1950 and 1960. Cumulative red bone marrow doses range up to 2 Gy with a mean of 0.3 Gy and a median of 0.2 Gy. Between 1953 and 2005, 93 first primary cases of leukemia, including 23 cases of chronic lymphatic leukemia (CLL), were ascertained among the cohort members. A significant linear dose-response relationship was seen for leukemias other than CLL (P < 0.001), but not for CLL. The estimated excess relative risk per Gy is 4.9 (95% confidence interval (CI): 1.6; 14.3) for leukemias other than CLL and less than 0 (95% upper bound 1.4) for CLL. PMID- 20012751 TI - Heart rate variability assessment in Japanese workers recovered from depressive disorders resulting from job stress: measurements in the workplace. AB - PURPOSE: The purpose of this study is to clarify workers' autonomic nerve balance after long-term sick leave due to depressive disorders resulting from job stress compared with healthy workers. METHODS: The participants were 28 Japanese male workers recovered from depressive disorders and 75 healthy male workers. For each participant, the lifestyle and the fatigue within 1 month were assessed by a checklist. Heart rate variability (HRV) was measured at the workplace by acceleration plethysmography (APG). HRV was assessed by the coefficient of variation of rate intervals (CV), the spectral components in the high- and low frequency areas represented by the normalized HF and LF (nHF and nLF), and the ratio of LF to HF components (LF/HF). RESULTS: There was no significant difference in individual lifestyle and fatigue symptoms between the recovered and the healthy workers. The former workers showed significantly lower CV, higher nLF and log(10)LF/HF, and lower nHF that represent the predominance of sympathetic activity in comparison with the healthy workers. Moreover, the recovered workers who discontinued medications indicated significantly higher nLF and log(10)LF/HF, and lower nHF compared to the recovered workers who continued their medications. CONCLUSIONS: Recovered workers in the workplace tended to show the depressive HRV feature that is the dominant sympathetic activity compared with the healthy workers. They might still be showing job stress that was not detected by the checklist. HRV analyses by APG in addition to questionnaire has the potential to become an effective approach for assessing workers' job stress to prevent repeated absences. PMID- 20012753 TI - Heat waves and heat days in an arid city in the northwest of Mexico: current trends and in climate change scenarios. AB - The aim of this work is to study heat waves (HWs) in Mexicali, Mexico, because numerous deaths have been reported in this city, caused by heatstroke. This research acquires relevancy because several studies have projected that the health impacts of HWs could increase under various climate change scenarios, especially in countries with low adaptive capacity, as is our case. This paper has three objectives: first, to analyze the observed change in the summer (1 June to 15 September) daily maximum temperature during the period from 1951 to 2006; secondly, to characterize the annual and monthly evolution of frequency, duration and intensity of HWs; and finally, to generate scenarios of heat days (HDs) by means of a statistical downscaling model, in combination with a global climate model (HadCM3), for the 2020 s, 2050 s, and 2080 s. The results show summer maximum temperatures featured warming and cooling periods from 1951 until the mid 1980s and, later, a rising tendency, which prevailed until 2006. The duration and intensity of HWs have increased for all summer months, which is an indicator of the severity of the problem; in fact, there are 2.3 times more HWs now than in the decade of the 1970s. The most appropriate distribution for modeling the occurrence of HDs was the Weibull, with the maximum temperature as co-variable. For the 2020 s, 2050 s, and 2080 s, HDs under a medium-high emissions scenario (A2) could increase relative to 1961-1990, by 2.1, 3.6, and 5.1 times, respectively, whereas under a medium-low emissions scenario (B2), HDs could increase by 2.4, 3.4, and 4.0, for the same projections of time. PMID- 20012752 TI - The role of surgery in clinical management of patients with metastatic papillary renal cell carcinoma. AB - OBJECTIVES: Patients with metastatic papillary renal cell carcinoma (RCC) show special clinical behavior compared to patients with other histologic subtypes of RCC. This study aimed to assess the relevance of surgical and systemic options used in treatment of these patients prior to the recent era of targeted therapies. METHODS: Retrospectively, we assessed clinical data of 61 patients with metastatic papillary RCC who were treated at eight centers in Germany. RESULTS: Median follow-up was 20 (range 1-114) months and median age at time of diagnosis was 62 (range 24-85) years. Men were affected predominantly (50/61; 82%). Twenty-one patients (34%) showed metastases at time of diagnosis. In the remaining 40 patients, median time to development of metastases was 30.4 (range 3 143; mean 16.5) months. Sites of metastases were lung (37; 61%), bone (24; 38%), liver (20; 33%), lymph nodes (24; 38%), and local recurrence (17; 28%). Others sites of disease were brain metastases (6 patients/10%), peritoneal carcinosis (5 patients/8%), and others. A surgical approach with potentially curative intention was performed primarily in 11 patients (18%). 31 patients received an immuno- (interferon-alpha +/- interleukin-2) or immunochemotherapy as first line treatment for metastatic disease. Overall, 42/61 patients (69%) received systemic therapy. Supportive care only was performed in 12 patients (20%) because of poor performance status. Median overall survival after diagnosis of metastatic disease was longer than 48 months in patients with tumor resection (n = 11) compared to 13.0 +/- 4.3 months 95% CI 4.5-21.5 (n = 42) months in patients without surgical approach. CONCLUSIONS: Complete resection of metastases represents a valid option in management of patients with relapsing or metastatic papillary RCC. PMID- 20012754 TI - Schmorl's nodes distribution in the human spine and its possible etiology. AB - Although Schmorl's nodes (SNs) are a common phenomenon in the normal adult population, their prevalence is controversial and etiology still debatable. The objective was to establish the spatial distribution of SNs along the spine in order to reveal its pathophysiology. In this study, we examined 240 human skeleton spines (T4-L5) (from the Hamann-Todd Osteological Collection) for the presence and location of SNs. To determine the exact position of SNs, each vertebral body surface was divided into 13 zones and 3 areas (anterior, middle, posterior). Our results show that SNs appeared more frequently in the T7-L1 region. The total number of SNs found in our sample was 511: 193 (37.7%) were located on the superior surface and 318 (62.3%) on the inferior surface of the vertebral body. SNs were more commonly found in the middle part of the vertebral body (63.7%). No association was found between the SNs location along the spine and gender, ethnicity and age. This study suggests that the frequency distribution of SNs varies with vertebra location and surface. The results do not lend support to the traumatic or disease explanation of the phenomenon. SNs occurrences are probably associated with the vertebra development process during early life, the nucleus pulposus pressing the weakest part of the end plate in addition to the various strains on the vertebrae and the intervertebral disc along the spine during spinal movements (especially torsional movements). PMID- 20012756 TI - Morpho-physiological and biochemical responses in the floating lamina of Trapa natans exposed to molybdenum. AB - The response to molybdenum (Mo) was studied in the metal-tolerant hydrophyte Trapa natans L. Previously, it was shown that the plant accumulates Mn in the floating lamina by means of phenolic compounds and responded with acclimation responses of the chloroplast. Since the involvement of phenolics has been proposed also in Mo resistance, we tested the response of T. natans to increasing doses (5, 50, 150, 600 microM) of Mo using the photosynthetic apparatus as an indicator of cellular stress. Only 5 microM Mo did not cause evident modifications with respect to controls. Conversely, 50 to 600 microM Mo induced progressively marked alterations of the lamina morphology. The chloroplast ultrastructure showed disorganisation of the thylakoid system, and correspondingly, the photosynthetic pigment pattern was altered with a fall-down in photosynthesis. Microspectrofluorimetry indicated alterations of photosystem II, with differences among the three cell layers (first and second palisade and spongy tissues). While the highest dose caused plant death, 50 and 150 microM Mo treated plants underwent partial recovery, and the plant survived up to the end of the vegetative season. However, reproduction was unsuccessful. Mo treatment did not induce increase in total phenolics, but only in anthocyanin. In contrast to Mn, detoxification of Mo by chelation inside vacuoles, possibly by anthocyanins, is suggested to be an insufficient mechanism to reduce Mo toxicity, which probably includes an impairment of nitrogen metabolism. However, the metal was accumulated in the lamina. On the whole, T. natans showed limited capabilities to survive Mo excess as compared with Mn. PMID- 20012755 TI - Lumbar posterolateral fusion inhibits sensory nerve ingrowth into punctured lumbar intervertebral discs and upregulation of CGRP immunoreactive DRG neuron innervating punctured discs in rats. AB - Degeneration of lumbar intervertebral discs is thought to be a cause of low back pain. Studies have found that a cause of discogenic low back pain is intervertebral disc inflammation and axonal growth of afferent fibers innervating the disc. Lumbar spine fusion for chronic discogenic low back pain is considered an effective procedure. However, no study has investigated the mechanism of pain relief. We did this by applying Fluoro-Gold (FG) to the ventral aspect of the L4 L5 intervertebral discs of 40 rats. We exposed the nucleus pulposus to the annulus fibrosus in a disc punctured model. Rats were divided into 4 groups. Group A: Punctured intervertebral disc with sham posterolateral fusion (PLF) (n = 10), Group B: Punctured intervertebral disc with PLF (n = 15), Group C: Normal intervertebral disc (no puncture) with PLF (n = 10), and Group D: Normal disc (no disc puncture) with sham PLF (n = 5). Four weeks after surgery, bilateral L1-L5 dorsal root ganglia (DRGs) were stained with growth-associated protein 43 (GAP43), a marker of axonal growth, and calcitonin gene-related peptide (CGRP), a neuropeptide marker of pain. Bone union was evaluated using X-ray imaging. Of the FG-labeled neurons, the proportions of GAP43- and CGRP-immunoreactive (IR) neurons in Group A were significantly higher than in Group D (P < 0.05). The proportions of GAP43- and CGRP-IR neurons in bone union rats in Group B were significantly lower than in nonunion rats in Group B and in the rats in Group A (P < 0.05). No significant differences in GAP43- and CGRP-IR neurons were observed between bone union and nonunion rats in Group C and the rats in Group D (P > 0.05). PLF is strongly related to the downregulation of GAP43 and CGRP expression. Therefore, PLF may suppress the increase of inflammatory neuropeptides and the process of axonal growth. Moreover, these results may explain, in part, the mechanism of pain relief following lumbar spinal fusion for chronic discogenic low back pain in humans. PMID- 20012757 TI - NADPH-diaphorase activity in the nociceptive pathways of land snail Megalobulimus abbreviatus: the involvement of pedal ganglia. AB - NADPH-diaphorase (NADPH-d) is a histochemical marker for nitric oxide synthase (NOS), widely used to identify nitric oxide (NO) producing cells in the nervous system of both vertebrates and invertebrates. Using NADPH-d histochemistry and semi-quantitative optical densitometry, we characterized the NO-producing neurons in the pedal ganglia of young and adult Megalobulimus abbreviatus, subjected to aversive thermal stimulus. The animals were killed at different times (3, 6, 12 and 24 h) following stimulus. The enzymatic activity was detected in different cellular subsets and neuronal processes. In all the studied pedal ganglia subregions, the optical density of positive neurons (P < 0.05) and neuropilar area 1 (P < 0.01) was significantly different in treated animals when compared to controls. The increase in nitrergic activity induced by nociceptive stimulus suggests the involvement of NO in the nociceptive circuit of M. abbreviatus, which is well maintained throughout evolution, and could be helpful in drawing cellular homologies with other gastropods. PMID- 20012758 TI - Histamine intolerance: a metabolic disease? AB - OBJECTIVE: To evaluate the evidence regarding the disease concept of histamine intolerance as a state of inadequate histamine inactivation. METHODS: Keyword based systematic screening of the scientific literature and of public websites focusing on diagnostic and therapeutic procedures. RESULTS: Histamine intolerance is commonly diagnosed based solely on subjective reporting of symptoms instead of following systematic diagnostic procedures based on objective laboratory and physical parameters. The only effective long-term therapy is avoidance of histamine-containing food. CONCLUSIONS: The concept of histamine intolerance as a metabolic disease is in need of more experimental and clinical evidence and affected patients will benefit from a clear, evidence-based diagnostic and therapeutic regime. PMID- 20012759 TI - Histamine in pericarditis of children with congenital heart malformations. AB - INTRODUCTION: Congenital heart malformations are risk factors that make children susceptible to infections resulting in inflammation. MATERIAL AND METHODS: The concentration of histamine as a modulator of inflammation was quantified in pericardial fluid and expression of histamine H(4) receptor (H(4)R) and histamine releasing factor (HRF) was determined at mRNA and protein levels. Samples of pericardium and pericardial fluid were obtained during cardiac reconstruction surgery in children. RESULTS: In children with pericarditis, increased levels of histamine were found and expression of H(4)R was localized on mast cells. Expression of HRF was independent of presence or absence of inflammation in pericardium and was localized within stationary epithelial cells. CONCLUSION: Results indicate that involvement of H4R in pericardial inflammation depends on penetration of mast cells into inflamed tissue, but HRF may not be directly involved in inflammatory reaction of the pericardium. PMID- 20012761 TI - On using time-averaging restraints in molecular dynamics simulation. AB - Introducing experimental values as restraints into molecular dynamics (MD) simulations to bias the values of particular molecular properties, such as nuclear Overhauser effect intensities or distances, (3)J coupling constants, chemical shifts or crystallographic structure factors, towards experimental values is a widely used structure refinement method. To account for the averaging of experimentally derived quantities inherent in the experimental techniques, time-averaging restraining methods may be used. In the case of structure refinement using (3)J coupling constants from NMR experiments, time-averaging methods previously proposed can suffer from large artificially induced structural fluctuations. A modified time-averaged restraining potential energy function is proposed which overcomes this problem. The different possible approaches are compared using stochastic dynamics simulations of antamanide, a cyclic peptide of ten residues. PMID- 20012760 TI - S3EPY: a Sparky extension for determination of small scalar couplings from spin state-selective excitation NMR experiments. AB - S3EPY is a Python extension to the program Sparky written to facilitate the assessment of coupling constants from in-phase/antiphase and spin-state-selective excitation (S(3)E) experiments. It enables the routine use of small scalar couplings by automating the coupling evaluation procedure. S3EPY provides an integrated graphical user interface to programs which outputs graphs and the table of determined couplings. PMID- 20012762 TI - Local mobility of 15N labeled biomolecules characterized through cross correlation rates: Applications to paramagnetic proteins. AB - The mobility of (15)N labeled proteins can be characterized by measuring the cross-correlation rates delta(N,NI) that govern the conversion of Zeeman order N(z) of an amide (15)N nucleus into longitudinal two-spin order 2N(z)I(z) involving the amide (15)N and (1)H nuclei. This represents an alternative to the measurement of (15)N self-relaxation rates 1/T(1) and 1/T(2) or 1/T(1rho). The rate of interconversion between N(z) and 2N(z)I(z) is due to cross-correlation between fluctuations of different interactions and is not affected by a variety of relaxation mechanisms that contribute to the self-relaxation rates 1/T(1), 1/T(2) and 1/T(1rho). Spin diffusion among protons, which affects the measurements, can be quenched by various means that are evaluated by experiments and simulations. By applying an off-resonance radio-frequency (RF) field in the vicinity of the nitrogen resonance, the spectral density function J(omega) can be determined at the frequency origin and at the nitrogen Larmor frequency. The methods are applied to the paramagnetic High-Potential Iron-Sulfur Protein iso I (HiPIP I) from E. halophila in its reduced state. PMID- 20012763 TI - Conformational analysis of protein and nucleic acid fragments with the new grid search algorithm FOUND. AB - The new computer algorithm FOUND, which is implemented as an integrated module of the DYANA structure calculation program, is capable of performing systematic local conformation analyses by exhaustive grid searches for arbitrary contiguous fragments of proteins and nucleic acids. It uses torsion angles as the only degrees of freedom to identify all conformations that fulfill the steric and NMR derived conformational restraints within a contiguous molecular fragment, as defined either by limits on the maximal restraint violations or by the fragment based DYANA target function value. Sets of mutually dependent torsion angles, for example in ribose rings, are treated as a single degree of freedom. The results of the local conformation analysis include allowed torsion angle ranges and stereospecific assignments for diastereotopic substituents, which are then included in the input of a subsequent structure calculation. FOUND can be used for grid searches comprising up to 13 torsion angles, such as the backbone of a complete alpha-helical turn or dinucleotide fragments in nucleic acids, and yields a significantly higher number of stereospecific assignments than the precursor grid search algorithm HABAS. PMID- 20012764 TI - PSEUDYANA for NMR structure calculation of paramagnetic metalloproteins using torsion angle molecular dynamics. AB - The program DYANA, for calculation of solution structures of biomolecules with an algorithm based on simulated annealing by torsion angle dynamics, has been supplemented with a new routine, PSEUDYANA, that enables efficient use of pseudocontact shifts as additional constraints in structure calculations of paramagnetic metalloproteins. PSEUDYANA can determine the location of the metal ion inside the protein frame and allows to define a single tensor of magnetic susceptibility from a family of conformers. As an illustration, a PSEUDYANA structure calculation is provided for a metal-undecapeptide complex, where simulated pseudocontact shifts but no NOE restraints are used as conformational constraints. PMID- 20012769 TI - Oxidative damage to DNA by 1,10-phenanthroline/L: -threonine copper (II) complexes with chlorogenic acid. AB - The oxidative DNA damage by copper (II) complexes in the presence of chlorogenic acid was explored using agarose gel electrophoresis. The extent of pBR322 DNA damage was enhanced significantly with increasing concentration of [Cu-phen-Thr] complex and incubation time. A fluorescence quenching activity of calf thymus DNA EB was observed more remarkably with chlorogenic acid than without chlorogenic acid. The fluorescence measurements suggested that [Cu-phen-Thr] complex not only can bind to DNA by intercalation but also can damage the double strand DNA in the presence of chlorogenic acid. Further, 8-hydroxy-2'-deoxyguanosine, a biomarker of DNA oxidative damage was determined by electrochemical method. The control experiments revealed that the structure of copper (II) complexes affected capability of complex to DNA damage. The planar structure copper (II) complex showed high efficiency to DNA damage. The chlorogenic acid as biological reductant could improve copper (II) complex to DNA damage. A mechanism on [Cu phen-Thr] complex to DNA damage in the presence of chlorogenic acid was proposed. PMID- 20012770 TI - The 22nd European Conference on Biomaterials: retrospective view, facts and figures. PMID- 20012771 TI - Hydroxyapatite bone substitutes developed via replication of natural marine sponges. AB - The application of synthetic cancellous bone has been shown to be highly successful when its architecture mimics that of the naturally interconnected trabeculae bone it aims to replace. The following investigation demonstrates the potential use of marine sponges as precursors in the production of ceramic based tissue engineered bone scaffolds. Three species of natural sponge, Dalmata Fina (Spongia officinalis Linnaeus, Adriatic Sea), Fina Silk (Spongia zimocca, Mediterranean) and Elephant Ear (Spongia agaricina, Caribbean) were selected for replication. A high solid content (80 %wt), low viscosity (126 mPas) hydroxyapatite slurry was developed, infiltrated into each sponge species and subsequently sintered, producing a scaffold structure that replicated pore architecture and interconnectivity of the precursor sponge. The most promising of the ceramic tissue engineered bone scaffolds developed, Spongia agaricina replicas, demonstrated an overall porosity of 56-61% with 83% of the pores ranging between 100 and 500 microm (average pore size 349 microm) and an interconnectivity of 99.92%. PMID- 20012772 TI - In vitro corrosion resistance and cytocompatibility of nano-hydroxyapatite reinforced Mg-Zn-Zr composites. AB - Due to good biocompatibility and mechanical properties, magnesium (Mg) and its alloys are considered promising degradable materials for orthopedic applications. In this work, a Mg metal matrix composite (MMC) was fabricated using Mg-2.9Zn 0.7Zr alloy as the matrix and 1 wt% nano-hydroxyapatite (n-HA) particles as reinforcements. In vitro corrosion behavior and cytocompatibility of a Mg-Zn-Zr/n HA composite and a Mg-Zn-Zr alloy were investigated. In contrast with the Mg-Zn Zr alloy, the MMC has better properties. The average corrosion rate of MMC is 0.75 mm/yr after immersion in simulated body fluid (SBF) for 20 days, and the surface of MMC is covered with white Ca-P precipitates. The electrochemical test results show that the corrosion potential (E(corr)) of MMC increases to -1.615 V and its polarization resistance (R(p)) is 2.56 KOmega with the addition of n-HA particles. The co-cultivation of MMC with osteoblasts results in the adhesion and proliferation of cells on the surface of the composite. The maximum cell density is calculated to be (1.85+/-0.15) x 10(4)/l after 5 days of co-culture with osteoblasts. The average cell numbers for two groups after culturing for 3 and 5 days (P<0.05) are significantly different. All the results demonstrate that the Mg-Zn-Zr/n-HA composite can be potentially used as biodegradable bone fixation material. PMID- 20012773 TI - Correlation of hydrolytic degradation with structure for copolyesters produced from glycolic and adipic acids. AB - Copolyesters based on glycolic acid (G) combined with adipic acid (A) and ethylene glycol (E) were synthesized in different percentage of molar ratios (A: 100-50% and G: 100%) and their hydrolytic degradation was studied and correlated with their structures. According to the DSC, the production of polyesters leads to the formation of copolyesters and not to mixtures of homopolyesters. The crystallites in the copolyesters mainly consist of continuous sequences of ethylene adipate structural units. The hydrolytic degradation of the polyesters was followed by their weight loss during hydrolysis and by the FTIR spectra of the initial polyesters compared with that of the degraded polyesters at equilibrium. The region between 1142 and 800 cm(-1) can be utilized to evaluate the extent of degradation of polyesters after their hydrolysis. The absorption bands at 1142, 1077 and 850 cm(-1) due to the amorphous region decrease after hydrolysis, whereas those at 972, 901 and 806 cm(-1) due to the crystalline region increase. The experimental data of the hydrolytic degradation were fitted with exponential rise to maximum type functions using two-parameter model, which describes very well mainly the initial part of the degradation, and four parameter model (containing two exponential terms), which is appropriate for fitting the hydrolytic degradation on the entire time period (including the equilibrium). Furthermore, the kinetics of the hydrolytic degradation of the polyesters for the initial time period based on both models results to similar values of the rate constant, k. The synthesized copolyesters of glycolic acid combined with adipic acid and ethylene glycol are soluble in many common organic solvents opposite to PGA, leading to modified biodegradable polyesters and therefore they can be easily processed. PMID- 20012775 TI - Occurrence and field densities of Coleoptera in the maize herb layer: implications for Environmental Risk Assessment of genetically modified Bt-maize. AB - Beetles (Coleoptera) are a diverse and ecologically important group of insects in agricultural systems. The Environmental Risk Assessment (ERA) of genetically modified Bt-crop varieties with insect resistances thus needs to consider and assess the potential negative impacts on non-target organisms belonging to this group. We analysed data gathered during 6 years of field-release experiments on the impact of two genetically modified Bt-maize varieties (Ostrinia-resistant MON810 and Diabrotica-resistant MON88017) on the occurrence and field densities of Coleoptera, especially the two families Coccinellidae and Chrysomelidae. Based on a statistical analysis aimed at establishing whether Bt-maize varieties are equivalent to their near-isogenic counterparts, we discuss the limitations of using field experiments to assess the effects of Bt-maize on these two beetle families. The densities of most of the beetle families recorded in the herb layer were very low in all growing seasons. Coccinellidae and Chrysomelidae were comparatively abundant and diverse, but still low in numbers. Based on their role as biological control agents, Coccinellidae should be a focus in the ERA of Bt plants, but given the large natural variability in ladybird densities in the field, most questions need to be addressed in low-tier laboratory tests. Chrysomelidae should play a negligible role in the ERA of Bt-plants, since they occur on-crop as secondary pests only. Species occurring off-crop, however, can be addressed in a similar fashion as non-target Lepidoptera in Cry1Ab expressing Bt-maize. PMID- 20012776 TI - Dynamics of a laterally evolving community of ribozyme-like agents as studied with a rule-based computing system. AB - The very early forms of life probably comprised ribozyme-like agents that were able to catalyze reactions and serve as templates for their own replication. The early evolution has also been suggested to occur mainly horizontally between proto-cells or inorganic compartments rather than vertically from parent cell to their dividing siblings. In order to study the evolutionary dynamics of such a community a rule-based computing system entitled as PrimordialEvo was developed. The system simulates a three dimensional matrix of compartments in which replicators, resource collectors and various other actors thrive. Horizontal movement between compartments may be due to genetically induced vesicle formation or random drift. Analysis of the simulation experiments suggests that active sharing of innovations between compartments is important for the overall reproductive success of life. The capability of natural selection to favor genes in the system was also tested, and, for example, the frequency of anti-parasites was observed to increase when parasites were allowed to emerge. PMID- 20012774 TI - Endotoxin exposure and lung cancer risk: a systematic review and meta-analysis of the published literature on agriculture and cotton textile workers. AB - OBJECTIVE: To examine the association between exposure to endotoxins and lung cancer risk by conducting a systematic review and meta-analysis of epidemiologic studies of workers in the cotton textile and agricultural industries; industries known for high exposure levels of endotoxins. METHODS: Risk estimates were extracted from studies published before 2009 that met predefined quality criteria, including 8 cohort, 1 case-cohort, and 2 case-control studies of cotton textile industry workers, and 15 cohort and 2 case-control studies of agricultural workers. Summary risk estimates were calculated using random effects meta-analyses. Potential sources of heterogeneity were explored through subgroup analyses. RESULTS: The summary risk of lung cancer was 0.72 (95% CI, 0.57-0.90) for textile workers and 0.62 (0.52-0.75) for agricultural workers. The relative risk of lung cancer was below 1.0 for most subgroups defined according to sex, study design, outcome, smoking adjustment, and geographic area. Two studies provided quantitative estimates of endotoxin exposure and both studies tended to support a dose-dependent protective effect of endotoxins on lung cancer risk. CONCLUSION: Despite several limitations, this meta-analysis based on high-quality studies adds weight to the hypothesis that occupational exposure to endotoxin in cotton textile production and agriculture is protective against lung cancer. PMID- 20012777 TI - Public involvement in pharmacogenomics research: a national survey on patients' attitudes towards pharmacogenomics research and the willingness to donate DNA samples to a DNA bank in Japan. AB - To assess the attitude of Japanese patients towards pharmacogenomics research and a DNA bank for identifying genomic markers associated with adverse drug reactions (ADRs) and their willingness to donate DNA samples, we conducted a survey of 550 male and female patients. The majority of the respondents showed a positive attitude towards pharmacogenomics research (87.6%) and a DNA bank (75.1%). The willingness to donate DNA samples when experiencing severe ADRs (55.8%) was higher than when taking medications (40.4%). Positive attitudes towards a DNA bank and organ donation were significantly associated with an increased willingness to donate. Though the level of positive attitude in the patient population was higher than that in the general public in our former study (81.0 and 70.4%, respectively), the level of the willingness of patients to donate was 40.4% when taking medications and 55.8% when experiencing severe ADRs which was lower than that of the general public in our former study (45.3 and 61.7%). The results suggested that the level of true willingness in the patient population was lower than that of the general public considering the fictitious situation presented to the public (to suppose that they were patients receiving medication). It is important to assess the willingness of patients who are true potential donors, not the general public. PMID- 20012784 TI - Absence of BRAF, NRAS, KRAS, HRAS mutations, and RET/PTC gene rearrangements distinguishes dominant nodules in Hashimoto thyroiditis from papillary thyroid carcinomas. AB - Dominant nodules within Hashimoto thyroiditis (HT) may present with unique morphological features that overlap with but are not diagnostic of papillary thyroid carcinoma (PTC). Activating BRAF point mutations, RAS aberrations, and RET rearrangements are mutually exclusive events in the oncogenesis of papillary thyroid carcinoma, and RET rearrangements have been previously described in dominant nodules of HT. We identified 28 cases of Hashimoto thyroiditis with a dominant nodule, from 345 consecutive HT thyroidectomies. Screening for BRAF, RET, KRAS, NRAS, and HRAS mutations, as well as RET-PTC1 and RET-PTC3 rearrangements, was performed on paraffin-embedded material from 17 of these dominant nodules. Patients ranged in age from 29 to 76 years and were predominantly female, and the nodules ranged from 1.5 to 6.2 cm. No BRAF or RAS mutations or RET-PTC rearrangements were identified in a dominant nodule, including those with atypical, worrisome histopathologic features. Of ten cases with diagnostic concomitant or incidental papillary carcinoma, three had a V600E point mutation in BRAF, and one case had a BRAF exon 15 deletion (600-604E), while the dominant nodules were negative for mutation, supporting the notion that dominant nodules are neither malignant nor precursor lesions, and strict histological, clinical, and molecular criteria must be met for the diagnosis of papillary thyroid carcinoma. PMID- 20012786 TI - Liver abscess in a newborn leading to portal vein thrombosis. AB - We report a male neonate who had liver abscess that resolved with intravenous antibiotics and surgical drainage. However, the child developed complete thrombosis of portal vein with cavernous formation within 16 days of therapy and portal hypertension subsequently. The child is now 2 1/2 years and has extra hepatic portal hypertension but is otherwise asymptomatic. PMID- 20012785 TI - Levosalbutamol vs racemic salbutamol in the treatment of acute exacerbation of asthma. AB - OBJECTIVE: To compare efficacy and tolerability of levosalbutamol (Group 1) and racemic salbutamol (Group 2) for the treatment of acute exacerbation of asthma in children age 5 to 18 yr. METHODS: A randomized double blind clinical study involving 60 children was undertaken between October' 06 to December' 07. RESULTS: The following baseline clinical characteristic were recorded initially and after giving 3 nebulizations at 20 min intervals in the Ist hour of presentation viz respiratory rate (RR), heart rate (HR), oxygen saturation in room air SPO2, PEFR (peak expiratory flow rate), serum K+ level and asthma score. In Group 1 patients (levosalbutamol), there was significant increment in SPO2 and PEFR (P<0.05) values with decrease in tachypnea and asthma score while no significant difference was found in pre and post treatment HR & Serum K+ levels. In Group 2 patients although there was clinical improvement in terms of SPO2, PEFR, RR and asthma score, it resulted in significant tachycardia and decrease in K+ levels. CONCLUSION: Levosalbutamol appears to be more efficacious than racemic salbutamol in terms of improvement in PEFR, SPO2 and asthma score while deleterious effects of tachycardia and fall in serum K+ were seen with racemic salbutamol. PMID- 20012787 TI - Familial small supernumerary marker chromosome (sSMC) (14)(:p11-q11:) [corrected] in a child with translocation Down syndrome. AB - We report a case of familial small supernumerary marker chromosome (sSMC)in a child with translocation Down syndrome (DS)and mother. The GTG-banded chromosomal analysis of DS child revealed 47,XY,+21,+mar and mother karyotype was 47,XX,+mar. The GTG-banded sSMC had a similar morphology of small acrocentric chromosomes. Fluorescence in situ hybridization (FISH)evaluation of sSMC using centromere probes(13/21,14/22,22)confirmed sSMC as derivative chromosome 14. The sSMC was not specifically stained with whole chromosome paint and arm-specific probes for chromosome 14;thus it has been described as der(14)(:p11-q11:).The phenotypic changes were not evident, may be due to trisomy condition in the child or the sSMC contain repetitive sequences. PMID- 20012788 TI - Diabetic ketoacidosis, thyroiditis and alopecia areata in a child with Down syndrome. AB - A 10-year-old girl with Down syndrome and alopecia areata was admitted with severe ketoacidosis. She had high blood glucose level: 615 mg/dL and her HbAlc level was 13.3 %. After the control of ketoacidosis and reduction of the plasma glucose level, we found low free T4 level and high TSH levels. Thyroid peroxidase antibodies titer was 1383 IU/mL and both gliadin and endomysial antibodies were negative. This was the first report of a child with Down syndrome showing type 1 diabetes combined with alopecia areata and autoimmune thyroid disease. PMID- 20012789 TI - Cyclic vomiting syndrome responding to amitryptiline. AB - A 7 year girl presented with history of recurrent vomiting and altered sensonium. Laboratory investigators were within normal limits. A diagnosis of cyclic vomiting syndrome was made and treated with amitryptiline with good results. PMID- 20012790 TI - Impact of attention-deficit/hyperactivity disorder on health-related quality-of-l ife of specific learning disability children [corrected]. AB - OBJECTIVE: To evaluate the impact of co-occurring attention-deficit/hyperactivity disorder (ADHD) on the health-related quality of life (HRQOL) of children with newly diagnosed specific learning disability (SpLD). METHODS: From September 2005 to March 2006, 150 parents (either mother or father) of children consecutively diagnosed as having SpLD were enrolled. The Child Health Questionnaire-Parent Form 50 (CHQ-PF50) was used to measure parent-reported HRQOL. CHQ-PF50 mean domain and summary scores computed for "SpLD/ADHD" and "SpLD only" children groups were compared using Independent Samples t-test. RESULTS: HRQOL of "SpLD/ADHD" children was significantly poorer in four domains: emotional impact on parents (mean difference: -11.0; 95% CI: -18.75--3.25; P = 0.006), general behavior (-9.61; 95% CI: -15.44--3.78; P = 0.002), time impact on parents (-9.51; 95% CI: -18.42--0.60; P = 0.038), and, self esteem (-7.62; 95% CI: -13.98--1.26; P = 0.020); and in overall psychosocial functioning (-5.34; 95% CI: -8.49--2.19; P = 0.001). CONCLUSION: Co-occurring ADHD adversely impacts the HRQOL of children with newly diagnosed SpLD especially in domains related to their psychosocial functioning. PMID- 20012791 TI - Fetal alcohol spectrum disorder(FASD. AB - Fetal alcohol syndrome (FAS)is the leading cause of mental retardation worldwide but is also the foremost preventable cause of neurobehavioral and developmental abnormalities. It is equally important to know spectrum of disorders due to maternal alcoholism during pregnancy such as Fetal alcohol spectrum disorder (FASD)in order to identify and treat affected child and family effectively. This article aims to create awareness among practising clinicians most of whom are only aware of phenotypical variant of FASD which is FAS.In this article we discuss those aspects of FASD relevant to the clinician such as: terminological ambiguity, assessment, diagnosis and prevention. PMID- 20012793 TI - Congenital dyserythropoietic anemia type I. PMID- 20012794 TI - Negative pressure pulmonary edema. PMID- 20012795 TI - Melamine-tainted milk: when China sneezes, Hong Kong catches cold. PMID- 20012796 TI - Routine axillary temperature monitoring in neonates cared under radiant warmer - is it necessary? PMID- 20012797 TI - Clinical and etiological profile of acute febrile encephalopathy in eastern Nepal. AB - OBJECTIVE: To investigate the clinical and etiological profile of acute febrile encephalopathy in children presenting to a tertiary care referral center of Eastern Nepal. METHODS: 107 children (aged 1 month to 14 yrs) presenting to the emergency with fever (> 38 C) of less than 2 weeks duration with altered sensorium with/or without seizure were prospectively investigated for etiological cause. The investigations included blood and CSF counts, blood and CSF cultures, peripheral smear and serology for malarial parasite, and serology for Japanese encephalitis (JE) virus. Other investigations included EEG and CT or MRI wherever indicated. RESULTS: The most common presenting complaints apart from fever and altered sensorium were headache and vomiting. Convulsions, neck rigidity, hypertonia, brisk deep tendon reflexes, extensor plantar response and focal neurological deficits were seen in 50%, 57%, 22.4%, 28%, 39.3% and 9.3% of the subjects, respectively. The diagnoses based on clinical presentation and laboratory findings were pyogenic meningitis in 45 (42%), non JE viral encephalitis in 26 (25%), JE in 19 (18%), cerebral malaria in 8 (7%), herpes encephalitis and tubercular meningitis in 4 (4%) each, and typhoid encephalopathy in 1 case. CONCLUSION: Pyogenic meningitis and viral encephalitis including JE are the most common causes of acute presentation with fever and encephalopathy. Preventive strategies must be directed keeping these causes in mind. PMID- 20012798 TI - Novel CHD7 and FBN1 mutations in an infant with multiple congenital anamolies. AB - The first case of an infant with a dual genetic diagnosis of CHARGE and Marfan syndrome is reported here. The patient had multiple congenital anamolies, many of them consistent with CHARGE syndrome and genetic testing identified a heterozygous mutation c.3806_11del6insA in the CHD7 gene. In addition, his father had physical features consistent with Marfan syndrome. Fibrillin-1 (FBN1) mutation screening identified a heterozygous c.3990insC mutation in both father and the patient. PMID- 20012799 TI - Investigation of the effect of high dairy diet on body mass index and body fat in overweight and obese children. AB - OBJECTIVE: To investigate whether an increase in dairy food consumption improves the changes in BMI and adiposity in children on an energy restricted diet. METHODS: Overweight and obese children (n = 120, age: 12-18 y, BMI: 27-40 kg/m2) were randomized to receive a calorie restricted diet providing a 500 kcal/d deficit from total energy expenditure and two (n = 40), three (n = 40) or four (n = 40) servings of dairy products/day. Anthropometric measurements in addition to serum hs-CRP and lipid profile were measured at baseline and after 12 weeks. RESULTS: Among the 96 children who completed the study, significant reductions in overall BMI, BMI z-score, weight, total body fat percentage and total body fat mass were observed (p < 0.001) but these reductions were not significantly affected by increasing dairy intake (p > 0.05). Overall waist/hip ratio, Serum vitamin D and lipid profile did not change significantly (p > 0.05) apart from a significant increase in HDL-cholesterol (p < 0.001) which was independent of dairy intake (p > 0.05). CONCLUSION: Increased intake of dairy products does not lead to an augmented change in BMI, weight and body fat in overweight and obese children beyond what is achieved by calorie restriction. PMID- 20012801 TI - Lung hypoplasia and its associated major congenital abnormalities in perinatal death: an autopsy study of 850 cases. AB - OBJECTIVE: To determine the relative frequency of causes of lung hypoplasia (LH) and its associated congenital malformations among perinatal deaths. METHODS: 850 medical reports of perinatal autopsies, in a 25-year period, assessed for LH as a cause of death. RESULTS: LH found in 96 (11.3%) cases, 89 (92.7%) were associated with major congenital malformation (secondary type) and primary type was seen in 7 cases (7.3%). Fourteen cases were associated with multiple congenital anomalies. 32 cases (33.3%) with Genito-urinary anomalies were the most common associated major malformations, followed by 19 cases (19.8%) of diaphragmatic impairment, 15 cases (15.6%) of musculoskeletal abnormalities and 11 cases (11.4%) of kidney agenesis. The most common musculoskletal abnormality was thanatophoric dwarfism in 10 cases (10.4%). Meckle-Gruber syndrome with 7 affected fetuses (7.3%) was the most common malformation syndrome associated with LH. CONCLUSION: More than ninety percent of LH was secondary to pathology outside the respiratory tract. Renal agenesis is the most common association observed in LH, followed by diaphragmatic hernia and thanatophoric dysplasia. PMID- 20012802 TI - Distal femoral duplication and fibular agenesis associated with congenital cardiac defect. AB - A newborn, who had the congenital anomalies including protuberance on the right lower leg, bilateral equinovarus deformity of the feet, genu valgum with knee flexion deformity, syndactyly between the first and the second digit on the right, in addition with the absence of the fifth digit. Echocardiography revealed a secundum type atrial septal defect. The combination of these congenital defects associated with developmental anomalies of lower extremities. We discuss the clinical, radiological findings and pathogenesis of this lower extremity malformation. PMID- 20012803 TI - Freeman-Sheldon syndrome--prenatal and postnatal diagnosis. AB - A six-day-old girl, born to normal non-consanguineous parents presented with mask like facies with a small mouth giving a 'whistling' appearance. Other dysmorphic features include deep set eyes, broad nasal bridge, long philtrum and 'H' shaped cutaneous dimple on the chin. There was congenital windmill vane hand position and severe talipes equinovarus deformity. The above features are characteristic of Freeman-Sheldon syndrome also known as Whistling Face syndrome. Ultrasound scanning during 8(th) month of the pregnancy showed the fetus to have facial abnormality and bilateral clenched hand and talipes with extension contractures of knees. Provisional diagnosis of FSS was made which was confirmed after the birth. Thus all cases of Arthrogryposis during prenatal scan should be carefully looked for the facial abnormality in the fetus. PMID- 20012804 TI - Turner syndrome and its variants. AB - Case records of female patients with karyotype proven turner syndrome were analyzed. 11 patients had classic Turner karyotype (Group 1) and 13 patients had karyotype suggestive of one of the variants of Turner syndrome (Group 2). There was a median difference of 3 years between the age of presentation and the age of diagnosis in Group 2. Out of the thirteen patients in Group 2, 4 had no clinical stigmata of Turner Syndrome; the rest (n=9) had one or more of the typical clinical stigmata of Turner Syndrome. One patient with a complex mosaic karyotype also had an intracranial medulloblastoma. One patient in each group had coarctation of the aorta. 5 patients in Group 1 and 3 patients in Group 2 had primary hypothyroidism and received levothyroxine. The median Thyroid Stimulating Hormone levels were significantly higher among patients in group 1 than in group 2. PMID- 20012806 TI - Pantoprazole-induced thrombocytopenia. PMID- 20012807 TI - Seckel syndrome with chromosomal 18 deletion. AB - Four case records of patients with Seckel Syndrome (SS) were retrieved. Typical of bird headed dwarfism was seen in all. Chromosome 18 deletion was seen in one child with SS. MRI abnormalities were detected in 3 patients. Cytogenetic studies and neuroimaging is likely to provide important diagnostic and prognostic information. PMID- 20012809 TI - A randomized trial of tailoring and motivational interviewing to promote fruit and vegetable consumption for cancer prevention and control. AB - BACKGROUND: Healthful dietary patterns, including eating fruits and vegetables (F&V) and avoiding obesity, may decrease the risk of cancer and other chronic diseases. In addition to promoting health for the general population, a cancer diagnosis may provide a "teachable moment," facilitating the adoption of more healthful eating habits and leading to lower risk of chronic disease and better overall health. PURPOSE: This study was designed to test the effectiveness of two health communication interventions in increasing F&V consumption and physical activity in a sample of older adults (average age of 66 years), including both colorectal cancer (CRC) survivors and noncolorectal cancer-affected (N-CRC) individuals. METHODS: CRC survivors and N-CRC individuals were recruited from a population-based case-control study and randomly assigned to four conditions using a 2 x 2 design. We tested two different methods of communicating and promoting health behavior change alone or in combination: tailored print communication (TPC) and brief telephone-based motivational interviewing (TMI). RESULTS: A significant increase in F&V consumption was found for the combined intervention group in the entire sample (p < 0.05). When stratified by cancer survivor status, the effect was concentrated in the N-CRC subset (p < 0.01) versus CRC survivors. The combined intervention was also found to be most cost effective for the N-CRC group, with TPC more cost-effective than TMI. For physical activity, none of the interventions produced statistically significant improvements. CONCLUSIONS: This study indicates that combining tailoring and motivational interviewing may be an effective and cost-effective method for promoting dietary behavior change among older healthy adults. More research is needed to identify the optimal dose and timing for intervention strategies to promote dietary and physical activity change among both CRC survivors and the general population. PMID- 20012810 TI - Mediators affecting girls' levels of physical activity outside of school: findings from the trial of activity in adolescent girls. AB - BACKGROUND: Providing after school activities is a community level approach for reducing the decline in physical activity of girls as they reach early adolescence. PURPOSE: The purpose of this study was to examine psychosocial, environmental, and behavioral factors as potential mediators of after school physical activity in adolescent girls. METHODS: We assessed objectively measured levels of physical activity occurring outside of school and potential predictors and mediators of activity in girls participating in the Trial of Activity in Adolescent Girls (TAAG). RESULTS: We found that the TAAG intervention had a statistically significant and positive effect on out of school activity in the 2006 cohort. Self-efficacy, friends' social support, total social support, and difficulty getting to and from community activities mediated the level of moderate to vigorous physical activity in girls. CONCLUSIONS: Parents, communities, and schools should provide and enhance opportunities outside of the school day for adolescents to be active. Reducing transportation barriers and enlisting social support appear to be key. PMID- 20012812 TI - Overview of antibodies for immunochemistry. AB - Immunohistochemistry is widely used to identify, in situ, various components of cells and tissues in both normal and pathological conditions and is an exceptionally powerful method to demonstrate the localization of cellular components. Immunoglobulins (antibodies) are glycoproteins and are divided into five major classes. IgG, which composes approximately 75% of the immunoglobulins in human serum, is most commonly used for immunostaining. Two types of detection systems, fluorescent and enzyme based are used for immunostaining. The choice of detection system depends on the type of sample and the availability of fluorescent or bight field microscopes as well as the type of information the investigator would like to obtain. This chapter provides an overview of antibody characteristics, and their use in immunostaining. PMID- 20012811 TI - Low life course socioeconomic status (SES) is associated with negative NEO PI-R personality patterns. AB - BACKGROUND: Low socioeconomic status (SES) is associated with poor health. One potential pathway accounting for this relationship may be an association between low SES and personality characteristics that affect health. METHODS: Associations among parent's education, current SES (education and income), and personality were examined among 233 African Americans and Caucasian, male and female community volunteers. RESULTS: Using multivariate analysis of variance (MANOVA) to model neuroticism, extraversion, openness to experience, agreeableness, and conscientiousness simultaneously, participant's education, household income, and father's and mother's education each had significant main effects on personality. When examining the life course--the combination of both current and childhood SES -distinctive patterns emerged for each domain, depending upon whether mother's or father's education was used to index childhood SES. When using mother's education as a childhood SES index, a high life course SES (high participant's SES/high mother's education) was associated with high extraversion and openness. Using father's education as a childhood SES index, a low life course SES (low participant's SES/low father's education) was associated with disproportionately high neuroticism and low conscientiousness. These effects did not differ by race or sex. CONCLUSION: The implications of these findings for the role of personality in the SES-health relationship are discussed. PMID- 20012813 TI - Introduction to the purification of antibodies. AB - Antibodies are a powerful and essential tool in scientific laboratories being used in an array of applications such as immuno-histochemistry, immunobloting, immunoprecipitation and enzyme-linked immunosorbent assays (ELISA). The different sources for antibodies include polyclonal antisera from immunized animals and monoclonal antibodies from cells in culture or from ascites in animals. Both polyclonal and monoclonal antibodies have their advantages, and or disadvantages, but in general the production of monoclonal antibodies is more time consuming and requires tissue culture facilities and skills. The use of either monoclonal or polyclonal antibodies in some of the applications may require that the antibody is in a purified form. They can be purified by a variety of methods described in the next few chapters. The availability of commercially available kits primarily designed for the purification of IgG and IgM classes of antibodies derived from all common animal species should also be mentioned. PMID- 20012814 TI - Antibody purification: ammonium sulfate fractionation or gel filtration. AB - Antibodies can be purified by a variety of methods based on their unique physical and chemical properties such as size, solubility, charge, hydrophobicity and binding affinity. This chapter focuses on ammonium sulfate precipitation as a convenient first step in antibody purification in that, it allows the concentration of the starting material and the precipitation of the desired protein. The principle of ammonium sulfate precipitation lies in "salting out" proteins from the solution. The proteins are prevented to form hydrogen bonds with water and the salt facilitates their interaction with each other forming aggregates that afterward precipitate out of solution. Gel filtration or size- exclusion chromatography is also discussed in this chapter. Gel filtration is based on the relative size of protein molecules and it is of great value to separate IgMs, exchange buffers and/or desalt solutions. The columns designed to separate the proteins are composed of porous beads and the proteins will flow through the packed column inside and around the beads, depending on its size. PMID- 20012815 TI - Antibody purification: ion-exchange chromatography. AB - Ion exchange chromatography techniques are the focus of this chapter and they showcase the power of this method for the purification of proteins and monoclonal antibodies. The technique is powerful and can separate biomolecules that have minor differences in their net charge, e.g., two protein molecules differing by a single charged amino acid. Given the amphoteric character of proteins the pH of the solution is important in the determination of the type of ion exchanger used. Immunoglobulins, although they can be purified by either cation or anion exchange chromatography, are most frequently purified by anion exchange with DEAE resins. The purification of the rabbit IgG fraction from serum using a DEAE column is detailed as well as the purification of IgG from ascitic fluid using FPLC, from loading to elution of the purified and concentrated protein. PMID- 20012816 TI - Antibody purification: affinity chromatography - protein A and protein G Sepharose. AB - Affinity chromatography relies on the reversible interaction between a protein and a specific ligand immobilized in a chromatographic matrix. The sample is applied under conditions that favor specific binding to the ligand as the result of electrostatic and hydrophobic interactions, van der Waals' forces and/or hydrogen bonding. After washing away the unbound material the bound protein is recovered by changing the buffer conditions to those that favor desorption. The technique has been used not only to isolate antigen-specific antibodies but also to remove specific contaminants from biological samples. Methods are described for the purification of immunoglobulins, namely IgG, IgG fragments and subclasses, using the high affinity of protein A and protein G coupled to agarose. In the Subheading 3 there are also protocols for affinity purification using a specific ligand coupled to commercial matrices like CNBr- Sepharose 4-B and Affigel. PMID- 20012817 TI - Conjugation of fluorochromes to antibodies. AB - Immunolocalization of antigen via fluorescence requires that fluorochromes be linked either to the primary antibody (direct method) or to a second antibody (indirect method) to provide a fluorescent signal to mark the site of antibody antigen binding. Of these two methods, the indirect technique is generally more useful and practical. Fluorochromes can be covalently conjugated to antibodies through reactions with thiol or amine groups. Typically, fluorochromes containing isothiocyanate, succinimidyl ester, or sulfonyl chloride reactive groups are conjugated to amines on the antibody molecules. Provided are step-by-step instructions for conjugating isothiocyanate derivates of fluorescein and sulfonyl chloride derivatives of rhodamine to the amine groups of antibodies. PMID- 20012818 TI - Biotinylation of antibodies. AB - Using the characteristic of a high-affinity complex between avidin and biotin, biotinylated antibodies have wide applications in various immunochemical assays, especially where signal amplification is required. A method is described here for the biotinylation of immunoglobulins. The procedure utilizes water-soluble succinimidyl ester of biotin that reacts with primary amines of the lysine residues or the amino terminus on the antibody to form amide bonds. The method is simple and specific and results in stable conjugates retaining full immunologic activity. PMID- 20012819 TI - Cell fixatives for immunostaining. AB - Fixation is one of the most critical steps in immunostaining. The object of fixation is to achieve good morphological preservation, while at the same time preserving antigenicity. Tissue blocks, sections, cell cultures or smears are usually immersed in a fixative solution, while in other situations, whole body perfusion of experimental animals is preferable. Fixation can be accomplished by either chemical or physical methods. The chemical methods include cross-linking agents such as formaldehyde, glutaraldehyde and succinimide esters as well as solvents such as acetone and methanol, which precipitate proteins. Of the physical methods, freezing tissue and air drying are most widely used. This chapter deals with the chemical fixation methods most commonly used for light microscopy. PMID- 20012820 TI - Permeabilization of cell membranes. AB - In order to detect intracellular antigens, cells must first be permeabilized especially after fixation with cross-linking agents such as formaldehyde and glutaraldehyde. Permeabilization provides access to intracellular or intraorganellar antigens. Two general types of reagents are commonly used: organic solvents, such as methanol and acetone, and detergents such as saponin, Triton X-100 and Tween-20. The organic solvents dissolve lipids from cell membranes making them permeable to antibodies. Because the organic solvents also coagulate proteins, they can be used to fix and permeabilize cells at the same time. Saponin interacts with membrane cholesterol, selectively removing it and leaving holes in the membrane. The disadvantage of detergents such as Triton X 100 and Tween-20 is that they are non-selective in nature and may extract proteins along with the lipids. This chapter provides methods for the use of organic solvents and detergents to permeabilize cell membranes. PMID- 20012821 TI - Preparation of frozen sections for analysis. AB - The analysis of frozen tissue by antibodies can be accomplished by the quick freezing of a small tissue sample in liquid nitrogen. Super-cooled isopentane can also be used to further the preservation process. Freezing preserves the available proteins in a near-native state for their identification by antibodies raised against naturally folded proteins. The tissues are sectioned onto charged glass slides where they can be optimally fixed in weakly or non-denaturing solutions such as acetone or those that are alcohol-based. Following mild pretreatment steps to allow for antibody use with low background, (the endogenous peroxidase enzyme or oxidative compounds quenched in a hydrogen peroxide solution and available charged sites blocked by incubation in a normal serum solution) the sections are ready for antigen detection. PMID- 20012822 TI - Processing of cytological specimens. AB - Individual cells often need to be examined with antibodies apart from the surrounding tissue. They may be cells in fluid, cells encased in mucus from a swab, or cells directly extracted from a piece of tissue. Cells can be viewed on a glass slide as cell smears produced from a cell enriched source, introduced as a touch preparation from a piece of wet tissue, concentrated on a slide by the use of a cytocentrifuge, or applied directly to a slide from a solid medium such as a cotton swab. These cell preparations can then be optimally fixed in weakly or nondenaturing solutions such as acetone or those that are alcohol based. They can also be postfixed in formalin if desired. Incubation in buffers containing 0.25% Triton X-100 and 5% dimethylsulfoxide (DMSO) allow for easier antibody penetration. The endogenous peroxidase enzyme or oxidative compounds can be quenched in a mild hydrogen peroxide solution. The sections are then ready to test with antibody after an incubation in a normal serum solution blocks any available charged sites. PMID- 20012823 TI - Processing of tissue culture cells. AB - Live cells are often studied, in vitro, bathed in nutrient growth media. It is sometimes necessary to study individual compounds produced by these cells and antibodies work well for this purpose. These cells must first be concentrated and fixed before testing. There are a couple of ways to study cells in culture using antibodies. One is to fix the cells in place as they adhere to a solid surface and then test them as though they were cells on a slide. Another is to retrieve them and pellet the cells, fixing them in a test tube and then embedding and sectioning them as though they were a solid tissue. Fixatives can be mild to moderate depending on the antigens to be studied. Sectioned cells can be tested following mild pretreatment steps. Cells fixed in the culture dish can be tested following mild pretreatment steps in buffers containing 0.25% Triton X-100 and 5% dimethylsulfoxide (DMSO) to allow for easier antibody penetration. The endogenous peroxidase enzyme or oxidative compounds can be quenched in a mild hydrogen peroxide solution. The sections are then ready to test with antibody after an incubation in a normal serum solution blocks any available charged sites. PMID- 20012824 TI - Processing of tissue specimens. AB - In order to test tissue specimens with antibody, they first have to be preserved in fixative, embedded in paraffin, and sectioned very thinly onto glass microscope slides. Any piece of tissue, immediately after excision, must be placed into an adequate volume of fixative. Fixatives vary, but the standard one is 10% buffered formalin. After an optimum fixation time (for formalin, about 16 h), the sample must be embedded in paraffin and sectioned on a microtome. Paraffin-embedded sections placed on positively charged slides (either coated or commercially prepared) are then ready for various pretreatment steps. First, the paraffin must be replaced with water through a series of rehydration steps. Then, depending on the antigen to be tested, the section can be proteolytically digested with enzymes or heat-treated in low or high pH solutions. Following that, the endogenous peroxidase enzyme or oxidative compounds can be quenched in a hydrogen peroxide solution. The sections are then ready to be tested with antibody after an incubation in a normal serum solution blocks any available charged sites. PMID- 20012825 TI - Heat-induced antigen retrieval for immunohistochemical reactions in routinely processed paraffin sections. AB - The development of heat-induced antigen (epitope) retrieval (HIER) technologies has led to dramatic improvements in our ability to detect antigens in formalin fixed, archival tissues. Paradoxically, wet heat treatment at temperatures greater than 95 degrees C in appropriate buffer solutions can reconstitute the antigenicity of many proteins that have been rendered nonreactive during the fixation and paraffin embedding process, which heretofore could only be identified in fresh or frozen tissues. The reason for this effect is unclear, but it has been suggested that the vigorous heat treatment partially reverses or disrupts the aldehyde cross-links occurring in proteins during formalin fixation and restores the original conformation of antigenic epitopes. The great success of antigen/epitope retrieval technologies further emphasizes the importance of preanalytical steps in immunohistochemistry. Over the past several years, since this technology was first reported, there have been numerous modifications to the original formulation. It is the purpose of this chapter to discuss the critical issues required for optimal HIER and to provide guidelines for the use of popular HIER buffers and heating devices used for routine immunohistochemical detection. PMID- 20012826 TI - Fluorochromes: properties and characteristics. AB - Immunofluorescence microscopy provides a sensitive means by which antigens can be localized within tissues or individual cells. For the most effective use of this technique the researcher can draw upon basic information on factors that affect the brightness of the fluorescence image, and how well that image can be distinguished from background fluorescence or interfering fluorescence signals. A wide variety of fluorochromes are available, with emitting wavelengths that range from the blue-violet end of the visible spectrum to the infrared. Individual fluorochromes are characterized by their extinction coefficients, quantum yields, susceptibility to photobleaching, the wavelengths at which they maximally absorb excitatory and emit fluorescent light, and how far apart those wavelength maxima are separated. Additional choices for fluorescent labeling of antibodies are provided by the availability of fluorescent quantum dots. Informed choices of fluorochromes can obviate many problems, particularly with regard to situations in which two or more antigens are to be localized simultaneously within a specimen. PMID- 20012827 TI - Direct immunofluorescent labeling of cells. AB - In the direct immunofluorescent labeling technique, fluorochrome-labeled antibodies are used as probes for particular antigens or biomolecules. Cells, usually after appropriate fixation, are incubated with the antibodies to which fluorochromes have been directly conjugated. Following incubation, excess antibody is washed off with PBS and the cells are mounted on coverslips with antifade mounting medium. Immunofluorescent labeled cells are analyzed using a conventional fluorescence microscope or by confocal microscopy. Direct labeling has two major advantages: it requires only a single incubation with the labeled reagent, decreasing the number of steps in the staining procedure; and more importantly, provides minimal nonspecific staining and less background. Additionally, the direct labeling technique allows the use of two or more primary antibodies of the same species or isotype, avoiding the problems with secondary antibody staining. This method has multiple applications: to label simultaneously two or more antigens within the same cell or tissue sections; to characterize the subcellular distribution of biomolecules of interest, by concurrently labeling with antibodies to both the antigen of interest and to a known organelle; to investigate whether several antigens of interest are colocalized; and to phenotype cells, for which no specific markers are available, using an appropriate panel of antibodies. PMID- 20012828 TI - Fluorescence labeling of surface antigens of attached or suspended tissue-culture cells. AB - This chapter deals with the detection of antigens that are accessible on the surface of isolated living cells using fluorochrome labels. By incubating live cells at 4 degrees C, to prevent endocytosis of bound molecules, the attached antibody can remain on the cell surface, and either be observed in the live state or subsequently fixed. This method yields the greatest sensitivity and best morphologic preservation for detection of surface molecules using antibodies for microscopy. PMID- 20012829 TI - Fluorescence labeling of intracellular antigens of attached or suspended tissue culture cells. AB - This chapter deals with the detection of antigens located in the interior of isolated cells, in which antibodies can detect their antigens only after fixation and permeabilization of cell structure. Unlike surface antigens, cell structure must be preserved first with a fixative, and the lipid barrier of the plasma membrane must be permeabilized using solvents or detergents. Fluorescence detection enhances the sensitivity of detection of small objects in the cytoplasm of cells because the fluorochrome acts as a point source of light. Because many fixatives preserve the native conformation of fixed antigens, it is important to select antibody reagents that can react with undenatured antigen. PMID- 20012831 TI - Overview of conventional fluorescence photomicrography. AB - In an age of digital imaging, photographic film still provides a viable and effective means for recording fluorescence images by photomicrography. To maximize the quality of results that are obtained, a photographic emulsion with sufficient sensitivity for the low light level characteristic of Immunofluorescence must be selected, exposures adjusted for reciprocity failure, and modern, high numerical aperture objective lenses employed to produce the brightest possible image. Mounting media that reduce the effects of photobleaching on fluorochromes also help to maintain image brightness, and so reduce exposure times. Digital scanning of film-based micrographs provides the convenience of utilizing image processing software to adjust image density and contrast, and to produce quality prints. PMID- 20012830 TI - Fluorescent visualization of macromolecules in Drosophila whole mounts. AB - The ability to determine the expression dynamics of individual genes "in situ" by visualizing the precise spatial and temporal distribution of their products in whole mounts by histochemical and immunocytochemical reactions has revolutionized our understanding of cellular processes. Drosophila developmental genetics was one of the fields that benefited most from these technologies, and a variety of fluorescent methods were specifically designed for investigating the localization of developmentally important proteins and cell markers during embryonic and post embryonic stages of this model organism. In this chapter we present detailed protocols for fluorescence immunocytochemistry of whole mount embryos, imaginal discs, pupal retinas, and salivary glands of Drosophila melanogaster, as well as methods for fluorescent visualization of specific subcellular structures in these tissues. PMID- 20012832 TI - Overview of confocal microscopy. AB - Born out of the need to overcome an imaging problem in the 1950s, confocal microscopes today allow researchers to go beyond simple imaging and ask biochemical questions. This chapter provides background information on the development of modern confocal microscopes, their uses and applications. Sample preparation and observation are also discussed. Information is also provided about more advanced applications such as FRAP, FRET and 2-photon imaging. The requirements for setting up a confocal laboratory and the instrumentation needs are also discussed. PMID- 20012833 TI - Overview of laser microbeam applications as related to antibody targeting. AB - This chapter reviews several techniques which combine the use of laser microbeams with antibodies to study molecular and cellular biology. An overview of the basic properties of lasers and their integration with microscopes and computers is provided. Biophysical applications, such as fluorescence recovery after photobleaching to measure molecular mobility and fluorescence resonance energy transfer to measure molecular distances, as well as ablative applications for the selective inactivation of proteins or the selective killing of cells are described. Other techniques, such as optical trapping, that do not rely on the interaction of the laser with the targeting antibody, are also discussed. PMID- 20012835 TI - Overview of antigen detection through enzymatic activity. AB - The identification of antigenic substances with antibodies can only occur through the use of a reporter molecule. One way of doing this is through the use of enzymes. Enzymes act upon a substrate and that substrate, or a molecule affected by that substrate, in turn becomes detectable by a variety of methods. There are many enzymes available for this purpose. The most common is peroxidase. Another widely used enzyme is alkaline phosphatase. Each enzyme has a few chromogenic substrate solutions with which it can react to change a color visualized through the use of selected instruments, including the microscope. Antibodies can be labeled with an enzyme directly, or secondary antibodies can be labeled with the enzyme and employed in an indirect technique. Also, immunoglobulin labeled polymers labeled with enzyme can be used and the enzymes themselves can serve as antigens in immunoenzyme complex procedures. Finally, avidin or biotin can be labeled with enzyme, and used either singly or in complexes, and peroxidase mediated biotin amplification can be used to increase the sensitivity in some procedures. PMID- 20012834 TI - Immuno-laser capture microdissection of rat brain neurons for real time quantitative PCR. AB - Laser capture microdissection (LCM) is a technical approach for obtaining microscopic samples as small as individual cells from tissues for molecular analysis. While the principles and details of the operation of LCM instruments, the technical requirements for obtaining identified cells for LCM "picking", all share the common feature of using a laser in combination with a microscope to microdissect and remove cells from tissue slices (or cultured cells) mounted on a glass slide. The use of LCM is becoming widespread in pathology laboratories and is increasingly being used for gene expression studies in cell biology. The approach is particularly powerful when used in conjunction with immunostaining techniques to obtain enriched RNA samples from cells that have been collected by picking and gathering phenotypically similar cells from anatomically complex organs such as the brain. In the present chapter, we describe an approach for combining immunocytochemistry with LCM to obtain RNA for real time quantitative PCR. PMID- 20012836 TI - The peroxidase-antiperoxidase (PAP) method and other all-immunologic detection methods. AB - Immunoenzyme procedures take on many forms, including, simply, antibody coupled to enzyme. These direct techniques require the labeling of all the primary antibodies and can produce more background. A more economical method uses a secondary antibody or one that has the primary antibody as its antigen. Labeling this secondary antibody with enzyme provides detection for many primary antibodies directed against different antigens of interest. A more sensitive approach involves the use of antibodies directed against enzyme connected to same species primary antibodies by a secondary linking antibody. This "all immunologic" technique is more sensitive and can result in less background than the covalently labeled methods. Finally, an immune polymer consisting of several secondary antibodies along with many enzyme molecules embedded on one long chain carbon polymer, can be used. This can provide a faster, more universal detection procedure with adequate sensitivity and specificity. PMID- 20012837 TI - The avidin-biotin complex (ABC) method and other avidin-biotin binding methods. AB - Immunoenzyme methods can be enhanced by the use of the high affinity molecules, avidin and biotin. The binding of avidin to biotin is almost irreversible. By labeling a detection enzyme such as horseradish peroxidase with biotin, and a secondary antibody (reactive against the antigen detecting primary antibody) with biotin as well, these two compounds can then be linked irreversibly with avidin. For this process, the biotinylated enzyme is complexed with avidin in solution and this avidin-biotin complex (ABC) is then introduced to the biotinylated secondary antibody, where it binds to primary antibody-antigen sites. Also, enzyme-labeled avidin molecules can be used to bind biotinylated secondary antibodies with greater resolution. Finally, biotinylated tyramide used in conjunction with peroxidase precipitates even greater amounts of biotin molecules for detection by enzyme-labeled avidin molecules. PMID- 20012838 TI - Avidin-biotin labeling of cellular antigens in cryostat-sectioned tissue. AB - Advances in immunohistochemical technologies have greatly improved the ability to visualize antigens in formalin-fixed paraffin-embedded tissues. Nonetheless, there are occasions in which there may be no alternative to the use of cryostat sectioned frozen tissues. While the basic chemistries and techniques used in staining cryostat-sectioned tissues are identical to those employed for staining paraffin-embedded sections, there are some unique issues to consider with frozen sections. Achieving excellent results requires the use of properly prepared and stored frozen tissue blocks embedded in an appropriate mounting compound, properly prepared and mounted cryostat-prepared sections, blocking of endogenous biotin and peroxidases that might interfere with interpretation of specific staining, and a mild postsectioning fixation step. This chapter describes a general frozen-section immunostaining procedure that provides guidelines for handling these special considerations. PMID- 20012839 TI - Multiple antigen immunostaining procedures. AB - Detection of multiple antigens in the same tissue section can be done by combining a range of immunohisto/cytochemical techniques based either on light microscopic chromogenic precipitates or fluorochrome labeling. Light microscopic techniques preferred for this purpose use combinations of immunogold silver staining (black precipitate), immunoperoxidase, immunoalkaline phosphatase and immunogalactosidase methods using chromogens of different colors. Fluorochrome labels favored for these combinations include AMCA (blue), FITC (green), rhodamine (orange-red) and Cy5 (far red), their matching synthetic members from the Alexa series, or quantum dots. Antibodies directly labeled or those from noncross-reacting animal species (e.g., mouse, rabbit, goat, guinea pig etc.) can be applied simultaneously. When the antigens of interest are in separate cells or cell compartments (e.g., in cell membrane, cytoplasm or nucleus), and only cross reacting antibodies are available, there have also been ways of avoiding unwanted cross-talk. These include the exploitation of the shielding effect of chromogens; inactivation of immuno-sequences of the first staining by using either acidic elution, formaldehyde fixation or microwave heating; combining unlabeled and hapten-labeled antibodies; or using labeled monovalent F(ab) secondary antibodies. In this chapter we briefly discuss the principle of multiple antigen immunolabeling and provide useful protocols for its performance. PMID- 20012840 TI - Immunoenzymatic quantitative analysis of antigens expressed on the cell surface (cell-ELISA). AB - Cell-enzyme-linked immunosorbent assay (cell-ELISA) is an useful technique for the quantitative analysis of cell surface antigen expression that was developed on the basis of enzyme immunohistochemistry (EIH) and ELISA. Since its development, which was made possible by the establishment of monoclonal antibody technology, a wide range of cell types and surface molecules were analyzed by cell-ELISA. Here we show four variants of this method and provide a brief comparison of cell-ELISA with flow cytometry (FACS) and radioimmunobinding assay (RIA), which are other methods for the quantitative detection of cell-surface molecules. We describe step-by-step procedures for both direct and indirect cell ELISA using either adherent or nonadherent live cells. PMID- 20012841 TI - Use of immunogold with silver enhancement. AB - Although gold particles are readily detectable by transmission electron microscopy, they can be difficult to visualize by bright-field light microscopy. However when the gold is silver-enhanced it is easy to see. During silver enhancement, the colloidal gold serves as a nucleation site for the deposition of metallic silver. The enhancing solutions are physical developers that contain both silver ions and a reducing agent, buffered to an acid pH. The silver enhancement method has also been used successfully to enlarge small-diameter gold particles for visualization by scanning electron microscopy. Silver-enhancement has been applied to a wide variety of tissues and antigens for both light and scanning electron microscopy. PMID- 20012842 TI - Overview of flow cytometry and fluorescent probes for flow cytometry. AB - This chapter provides an introduction to the use of fluorescent probes in flow cytometry. Sample preparation for the use of surface labeling with antibodies as well as for the use of nucleic acid probes is discussed. The utility of cell sorting is also discussed. PMID- 20012843 TI - Tissue disaggregation. AB - A conventional flow cytometry procedure for creating single cell suspensions is provided. Methods of tissue disaggregation to create single-cell suspensions from formalin fixed tissue are also given. Chemical, mechanical, and enzymatic multistep methods for separating single cells from tissue are discussed. These procedures are very tissue dependent. PMID- 20012844 TI - Indirect immunofluorescent labeling of viable cells. AB - A recognized flow cytometry procedure for standard indirect antibody staining for cellular analysis and cell sorting is discussed. A protocol for cell surface or intracellular antibody staining of viable cells is provided. PMID- 20012845 TI - Indirect immunofluorescent labeling of fixed cells. AB - Flow cytometry protocols for defining cell surface or intracellular antibody staining are discussed. Various staining protocols are provided. Routine cell surface and intracellular techniques as well as more advanced signal enhancement techniques are detailed. PMID- 20012846 TI - Fluorescent labeling of DNA. AB - A well-defined method for staining cellular DNA especially for cell cycle determination is provided. Emphasis is placed on utilizing DNA content and cell sizing measurement to further define cell populations. PMID- 20012847 TI - Deparaffinization and processing of pathologic material. AB - Methods for preparation of whole nuclei from paraffin embedded tissue are given. The combination of cell sorting of isolated nuclei combined with the polymerase chain reaction can be used to investigate other cellular parameters such as nuclear proteins and proliferation factors. PMID- 20012848 TI - Fixation and embedding. AB - For electron microscopic immunocytochemistry, the fixation procedure is always a compromise between good morphological preservation and retention of antigenicity. The choice of fixative depends on whether the immunogold labeling will be done before or after the samples are embedded and on how resistant the antigen is to fixation. For preembedding staining, it is possible to immunolabel the samples prior to fixation or after only a very mild fixation. Following immunolabeling, the samples can be refixed in a stronger fixative, such as 2% glutaraldehyde, to give good morphological preservation. Since it is not possible to refix the tissue after immunolabeling, for postembedding labeling, the composition of the initial fixative must be such that morphological detail and antigenicity are both preserved. While virtually any embedding resin may be used for immunogold staining, for postembedding methods, the resin can affect the immunostaining. In this chapter, methods are given for conventional fixation and microwave fixation as well as for embedding in various resins. PMID- 20012849 TI - Preparation of colloidal gold. AB - Colloidal gold probes have become widely used for immunocytochemical staining at the electron microscopic level. Gold sols are producing by boiling a solution of tetrachloroauric acid with a reducing agent. The type of reducing agent and the concentration of components determine the final particle size. Gold sols that have a particle size ranging from 2 to 40 nm can be made in the laboratory, depending on the type and concentration of the reducing agent. This chapter details methods for producing various sizes of gold. The methods are relatively simple and very reproducible from batch to batch. PMID- 20012850 TI - Conjugation of colloidal gold to proteins. AB - The ability to conjugate proteins to colloidal gold sols provides a wide variety of probes for electron microscopy. Antibodies, protein A, protein G, lectins, enzymes, toxins, and other proteins have all been conjugated to colloidal gold. The nature of the interaction between the colloidal gold and the protein is poorly understood. Proteins are conjugated to gold sols by adjusting the pH of the gold sol to approximately 0.5 pH unit higher than the pI of the protein being conjugated. This chapter gives a general method for conjugating proteins to colloidal gold as well as more specific methods for conjugating antibodies and protein A to colloidal gold. PMID- 20012851 TI - Colloidal gold/streptavidin methods. AB - Biotin-avidin detection systems are widely used in both immunocytochemistry and molecular biology. They take advantage of the high affinity of biotin, a low molecular-weight vitamin, for avidin, an egg-white protein. Because of the problem of nonspecific binding of avidin, streptavidin has largely replaced avidin for immunocytochemical procedures. Streptavidin/colloidal gold-biotin detection systems for electron microscopy are most commonly used in postembedding immunocytochemistry. Usually, the primary antibody is unlabeled, the secondary antibody is biotinylated, and the colloidal gold is conjugated to streptavidin. In certain applications, the primary antibody may be biotinylated and no bridging antibody is needed. This chapter details the use of streptavidin-gold for postembedding labeling. PMID- 20012852 TI - Pre-embedding labeling methods. AB - Colloidal gold conjugates generally do not readily penetrate cells, even after permeabilization. Therefore, their use in pre-embedding immunostaining has been largely restricted to labeling cell-surface antigens for scanning or transmission electron microscopy or for tracing endocytic pathways in living cells. One nanometer gold conjugates that do penetrate cells and tissues much more readily have also been used successfully to immunolabel intracellular structures. For pre embedding labeling, all of the immunostaining is done prior to embedding the tissue in resin or preparing the samples for scanning electron microscopy. This chapter provides methods for pre-embedding staining with unconjugated primary antibody or with primary antibody conjugated to colloidal gold. The use of colloidal gold for tracing endocytic pathways is also given. PMID- 20012853 TI - Postembedding labeling methods. AB - Since it was first introduced, postembedding immunogold labeling has become the most widely used method of immunolabeling for electron microscopy. For postembedding labeling, samples are first fixed, embedded, and sectioned. All immunostaining is performed on sections mounted on grids. The immunostaining may be done with a direct labeling technique with the primary antibody conjugated to colloidal gold, or by an indirect method where the primary antibody is unlabeled and the gold is conjugated to a secondary or tertiary antibody, protein A, protein G, etc. Colloidal gold-antibody conjugates are also widely used, especially in indirect immunocytochemical methods. This chapter gives the standard method for post embedding labeling as well as one in which the cell membranes are enhanced. PMID- 20012854 TI - The clinical immunohistochemistry laboratory: regulations and troubleshooting guidelines. AB - The Clinical Laboratory Improvement Amendments (CLIA) set standards designed to improve the quality of all laboratory testing. In the first portion of this chapter, we discuss the CLIA requirements that apply to most Immunohistochemistry laboratories, and explain topics such as certification, test complexity, patient test management, proficiency testing, personnel, quality control, quality assurance, and compliance. The second portion of this chapter addresses the most common problems encountered in immunohistochemical procedures and the appropriate solutions to correct them. PMID- 20012855 TI - Mapping quantitative trait loci for quality factors in an inter-class cross of US and Chinese wheat. AB - Wheat quality factors are critical in determining the suitability of wheat (Triticum aestivum L.) for end-use product and economic value, and they are prime targets for marker-assisted selection. Objectives of this study were to identify quantitative trait loci (QTLs) that ultimately influence wheat market class and milling quality. A population of 132 F(12) recombinant inbred lines (RILs) was derived by single-seed descent from a cross between the Chinese hard wheat line Ning7840 and the soft wheat cultivar Clark and grown at three Oklahoma locations from 2001 to 2003. Milling factors such as test weight (volumetric grain weight, TW), kernel weight (KW), and kernel diameter (KD) and market class factors such as wheat grain protein content (GPC) and kernel hardness index (HI) were characterized on the basis of a genetic map constructed from 367 SSR and 241 AFLP markers covering all 21 chromosomes. Composite interval mapping identified eight QTLs for TW, seven for KW, six for KD, two each for GPC and HI measured by near infrared reflectance (NIR) spectroscopy, and four for HI measured by single kernel characterization system. Positive phenotypic correlations were found among milling factors. Consistent co-localized QTLs were identified for TW, KW, and KD on the short arms of chromosomes 5A and 6A. A common QTL was identified for TW and KD on the long arm of chromosome 5A. A consistent major QTL for HI peaked at the Pinb-D1 locus on the short arm of chromosome 5D and explained up to 85% of the phenotypic variation for hardness. We identified QTLs for GPC on 4B and the short arm of 3A chromosomes. The consistency of quality factor QTLs across environments reveals their potential for marker-assisted selection. PMID- 20012856 TI - Regulation of seed yield and agronomic characters by photoperiod sensitivity and growth habit genes in soybean. AB - Soybean genotypes are adapted to narrow bands of latitude due to photoperiod sensitivity. There are several photoperiod-sensitive loci (E1, E2, E3, E4, E5, E6, E7, E8). Determinate and indeterminate growth habits are controlled by a single locus. The objective of our research was to examine the effects of photoperiod sensitivity and growth habit alleles on seed yield and other agronomic characters using isogenic lines. Twenty 'Harosoy' isolines with 11 photoperiod-sensitive genotypes many with both indeterminate and determinate growth habits were grown in the field at Ottawa, ON, from 2003 to 2007. Maturity ranged from 97 to 127 days, and seed yield increased linearly with maturity until about 112 days when it plateaued. Determinate lines were always shorter than indeterminate lines of equivalent maturity. Seed yield was associated with plant height, maturity, seed sugar concentration, seed weight and lodging. Effects of alleles at individual loci, and additive and epistatic effects across multiple loci were examined. At a single locus, photoperiod-insensitive alleles produced isolines that matured 8-11 days earlier, yielded less, and had shorter plants with reduced lodging. In multiple loci analyses, additive effects explained most of the variation in agronomic characters since additive models with E1, E3, E4, E7 and Dt1 loci included compared well to additive plus epistatic models and genotype-based models. Variation in photoperiod sensitivity and growth habit alleles results in a range of maturity, with pleiotropic effects on seed yield and agronomic characteristics, and play an important role in providing adaptation across latitudes. PMID- 20012857 TI - Effects of aerobic exercise, resistance exercise or both, on patient-reported health status and well-being in type 2 diabetes mellitus: a randomised trial. AB - AIMS/HYPOTHESIS: The Diabetes Aerobic and Resistance Exercise (DARE) study showed that aerobic and resistance exercise training each improved glycaemic control and that a combination of both was superior to either type alone in patients with type 2 diabetes mellitus. Here we report effects on patient-reported health status and well-being in the DARE Trial. METHODS: We randomised 218 inactive participants with type 2 diabetes mellitus in parallel to 22 weeks of aerobic exercise (n = 51), resistance exercise (n = 58), combined aerobic and resistance exercise (n = 57) or no exercise (control; n = 52). Intervention allocation was managed by a central office. Outcomes included health status as assessed by the physical and mental component scores of the Medical Outcomes Trust Short-Form 36 item version (SF-36) and well-being as measured by the Well-Being Questionnaire 12-item version (WBQ-12); these were measured at the Ottawa Hospital. RESULTS: Using a p value of 0.0125 for statistical significance due to multiple comparisons, mixed model analyses indicated that resistance exercise led to clinically but not statistically significant improvements in the SF-36 physical component score compared with aerobic exercise (Delta = 2.7 points; p = 0.048) and control (i.e. no exercise; Delta = 3.3 points; p = 0.015). For mental component scores, there were clinically important improvements favouring no (control) compared with resistance (Delta = 7.6 points; p < 0.001) and combined (Delta = 7.2 points; p < 0.001) exercise. No effects on WBQ-12 scores were noted. Overall, 59/218 (27%) of participants included in this analysis sustained an adverse event during the course of the study, including 16 participants in the combined exercise group, 19 participants in the resistance exercise group, 16 participants in the aerobic exercise group, and eight participants in the control group. All participants were included in the intent-to-treat analyses. The trial is now closed to follow-up. CONCLUSIONS/INTERPRETATION: Resistance exercise was better than aerobic or no exercise for improving physical health status in these patients. No exercise was superior to resistance or combined exercise for improving mental health status. Well-being was unchanged by intervention. TRIAL REGISTRATION: ClinicalTrials.gov NCT00195884 FUNDING: This study was funded by the Canadian Institutes of Health Research (grant MCT-44155) and the Canadian Diabetes Association (The Lillian Hollefriend Grant). PMID- 20012858 TI - Gut barrier disruption by an enteric bacterial pathogen accelerates insulitis in NOD mice. AB - AIMS/HYPOTHESIS: Increased exposure to enteric microbes as a result of intestinal barrier disruption is thought to contribute to the development of several intestinal inflammatory diseases; however, it less clear whether such exposure modulates the development of extra-intestinal inflammatory and autoimmune diseases. The goal of this study was to examine the potential role of pathogenic enteric microbes and intestinal barrier dysfunction in the pathogenesis of type 1 diabetes. METHODS: Using NOD mice, we assessed: (1) intrinsic barrier function in mice at different ages by measuring serum levels of FITC-labelled dextran; and (2) the impact on insulitis development of infection by strains of an enteric bacterial pathogen (Citrobacter rodentium) either capable (wild-type) or incapable (lacking Escherichia coli secreted protein F virulence factor owing to deletion of the gene [DeltaespF]) of causing intestinal epithelial barrier disruption. RESULTS: Here we demonstrate that prediabetic (12-week-old) NOD mice display increased intestinal permeability compared with non-obese diabetes resistant and C57BL/6 mice. We also found that young (4-week-old) NOD mice infected with wild-type C. rodentium exhibited accelerated development of insulitis in concert with infection-induced barrier disruption. In contrast, insulitis development was not altered in NOD mice infected with the non-barrier disrupting DeltaespF strain. Moreover, C. rodentium-infected NOD mice demonstrated increased activation and proliferation of pancreatic-draining lymph node T cells, including diabetogenic CD8(+) T cells, compared with uninfected NOD mice. CONCLUSIONS/INTERPRETATION: This is the first demonstration that a loss of intestinal barrier integrity caused by an enteric bacterial pathogen results in the activation of diabetogenic CD8(+) T cells and modulates insulitis. PMID- 20012859 TI - Prevalence and correlates of probable adolescent mental health problems reported by parents in Vietnam. AB - PURPOSE: The purpose of the present study was to estimate the prevalence of probable mental health problems in an epidemiologic study of Vietnamese adolescents. A secondary aim was to examine the correlates of probable mental health caseness. METHODS: Interviewers visited 1,914 households that were randomly selected to participate in a multi-agency study of mental health in select provinces of Vietnam. Semi-structured interviews assessed adolescent mental health problems using the Strengths and Difficulties Questionnaire (SDQ) parent informant version, and additionally the interviewers collected information on demographic variables (age, gender, ethnic group, religious affiliation, social capital). The final sample included data on 1,368 adolescents (aged 11-18 years). RESULTS: The average score on the total problem composite of the SDQ scale was 6.66 (SD=4.89), and 9.1% of the sample was considered a case (n=124). Bivariate analyses were conducted to determine which demographic variables were related to the SDQ case/non-case score. All variables except gender were significant in bivariate analyses, and therefore were entered into a logistic regression. Results indicated that age, religion, and wealth remained significant predictors of probable caseness. CONCLUSIONS: Overall, prevalence estimates of mental health problems generated by the SDQ were consistent with those reported in the US and other Western and non-Western samples. Results of the current study suggest some concordance of risk and protective factors between Western and Vietnamese youth (i.e., age and SES). PMID- 20012860 TI - IL-17E, a proinflammatory cytokine, has antitumor efficacy against several tumor types in vivo. AB - Interleukin-17E (IL-17E) belongs to a novel family of cytokines that possess significant homology to IL-17. IL-17E has potent inflammatory effects in vitro and in vivo. Overexpression of IL-17E in mice results in a T helper-2 (Th2)-type immune response, which includes the expansion of eosinophils through the production of IL-5, and elevated gene expression of IL-4 and IL-13 in multiple tissues. In this study, we show that IL-17E has antitumor activity in vivo, a previously unrecognized function of IL-17E. Antitumor efficacy of IL-17E was examined in a variety of human tumor xenograft models, including melanoma, breast, lung, colon, and pancreatic cancers. Injection of recombinant IL-17E every other day resulted in significant antitumor activity in these tumor models. In addition, the combination of IL-17E with chemotherapy or immunotherapy agents showed an enhanced antitumor efficacy in human tumor xenograft models in mice as compared to either agent alone. Antitumor activity was demonstrated using different routes of administration, including intraperitoneal, intravenous, and subcutaneous injection. Anticancer activity was shown for both mouse and human forms of IL-17E, which have a high degree of sequence identity. Tumor-bearing mice treated with IL-17E showed a significant increase in serum levels of IL-5 and increased numbers of eosinophils in peripheral blood compared to the control group. Spleens isolated from IL-17E-treated mice showed a significant increase in eosinophils that correlated with antitumor activity of IL-17E in a dose-response manner. Finally, we demonstrate that B cells are necessary for IL-17E-mediated antitumor activity and that IL-17E was found to activate signaling pathways in B cells in vitro. Taken together, these data demonstrate that IL-17E has antitumor activity in vivo, and support further investigation of the potential clinical use of IL-17E as an anticancer agent. PMID- 20012861 TI - The impact of pathological fractures on therapy outcome in patients with primary malignant bone tumours. AB - The primary objective of this study was to investigate the implications of pathological fractures on therapy outcome in patients with primary malignant bone tumours and to determine whether limb salvage can be safely performed. A retrospective analysis of 447 patients with primary malignant bone tumours, treated between 1985 and 2005, was performed. Multivariate Cox regression analysis was used to investigate the influence of pathological fractures and further independent variables on survival rate. In 52 of the 447 patients, the primary malignant bone tumour was complicated by a pathological fracture. These fractures were more common in malignant fibrous histiocytoma (MFH) of the bone and in the tumour stages IIa/b and III. Ablative surgery was performed in ten patients and limb salvage surgery in 42 patients. The mortality risk for patients with pathological fractures was significantly increased by a factor of 1.82 (p = 0.015), and overall duration of survival was significantly lower in the fracture group, with a median of 6.2 years (p < 0.00001). In univariate and multivariate analysis, fracture, higher tumour stages and resection margins remained a significant predictor of worse survival. Overall survival, rate of local recurrence and distant metastases were not affected by the type of surgical treatment selected; there was no difference between the patients who underwent limb salvage and those who underwent an amputation. Pathological fracture in patients with primary malignant bone tumours is a predictor of worse survival and significantly increases mortality risk. Reconstructive surgery did not influence the survival rate, showing that limb salvage therapy is safe when adequate resection margins are achieved. PMID- 20012862 TI - Early migration characteristics of a hydroxyapatite-coated femoral stem: an RSA study. AB - Measurement of early stem subsidence can be used to predict the likelihood of long-term femoral component loosening and clinical failure. Data that examines the early migration pattern of clinically proven stems will provide clinicians with useful baseline data with which to compare new stem designs. This study was performed to evaluate the early migration pattern of a hydroxyapatite-coated press-fit femoral component that has been in use for over ten years. We enrolled 30 patients who underwent THA for osteoarthritis. The median age was 70 years (range, 55-80 years). Patients were clinically assessed using the Harris hip score. Radiostereometric analysis was used to evaluate stem migration at three to four days, six months, one year and two years. We observed a mean subsidence of 0.73 mm at six months, 0.62 mm at one year and 0.58 mm at two years and a mean retroversion of 1.82 degrees at six months, 1.90 degrees at one year and 1.59 degrees at two years. This data suggests that subsidence is confined to the first six months after which there was no further subsidence. The results from this study can be compared with those from novel cementless stem designs to help predict the long-term outcome one may expect from new cementless stem designs. PMID- 20012863 TI - A novel, small molecule inhibitor of Hsc70/Hsp70 potentiates Hsp90 inhibitor induced apoptosis in HCT116 colon carcinoma cells. AB - PURPOSE: The anti-apoptotic function of the 70 kDa family of heat shock proteins and their role in cancer is well documented. Dual targeting of Hsc70 and Hsp70 with siRNA induces proteasome-dependent degradation of Hsp90 client proteins and extensive tumor specific apoptosis as well as the potentiation of tumor cell apoptosis following pharmacological Hsp90 inhibition. METHODS: We have previously described the discovery and synthesis of novel adenosine-derived inhibitors of the 70 kDa family of heat shock proteins; the first inhibitors described to target the ATPase binding domain. The in vitro activity of VER-155008 was evaluated in HCT116, HT29, BT474 and MDA-MB-468 carcinoma cell lines. Cell proliferation, cell apoptosis and caspase 3/7 activity was determined for VER 155008 in the absence or presence of small molecule Hsp90 inhibitors. RESULTS: VER-155008 inhibited the proliferation of human breast and colon cancer cell lines with GI(50)s in the range 5.3-14.4 microM, and induced Hsp90 client protein degradation in both HCT116 and BT474 cells. As a single agent, VER-155008 induced caspase-3/7 dependent apoptosis in BT474 cells and non-caspase dependent cell death in HCT116 cells. VER-155008 potentiated the apoptotic potential of a small molecule Hsp90 inhibitor in HCT116 but not HT29 or MDA-MB-468 cells. In vivo, VER 155008 demonstrated rapid metabolism and clearance, along with tumor levels below the predicted pharmacologically active level. CONCLUSION: These data suggest that small molecule inhibitors of Hsc70/Hsp70 phenotypically mimic the cellular mode of action of a small molecule Hsp90 inhibitor and can potentiate the apoptotic potential of a small molecule Hsp90 inhibitor in certain cell lines. The factors determining whether or not cells apoptose in response to Hsp90 inhibition or the combination of Hsp90 plus Hsc70/Hsp70 inhibition remain to be determined. PMID- 20012864 TI - Histone modifying proteins Gcn5 and Hda1 affect flocculation in Saccharomyces cerevisiae during high-gravity fermentation. AB - The performance of yeast is often limited by the constantly changing environmental conditions present during high-gravity fermentation. Poor yeast performance contributes to incomplete and slow utilization of the main fermentable sugars which can lead to flavour problems in beer production. The expression of the FLO and MAL genes, which are important for the performance of yeast during industrial fermentations, is affected by complex proteins associated with Set1 (COMPASS) resulting in the induction of flocculation and improved maltose fermentation capacity during the early stages of high-gravity fermentation. In this study, we investigated a possible role for other histone modifying proteins. To this end, we tested a number of histone deacetylases (HDACs) and histone acetyltransferases and we report that flocculation is induced in absence of the histone deacetylase Hda1 or the histone acetyltransferase Gcn5 during high-gravity fermentation. The absence of Gcn5 protein also improved utilization of high concentrations of maltose. Deletion of SIR2 encoding the HDA of the silent informator regulator complex, did not affect flocculation under high-gravity fermentation conditions. Despite the obvious roles for Hda1 and Gcn5 in flocculation, this work indicates that COMPASS mediated silencing is the most important amongst the histone modifying components to control the expression of the FLO genes during high-gravity fermentation. PMID- 20012865 TI - A clinical picture of chronic polyarticular tophaceous gout. AB - An unusual picture of polyarticular tophaceous gout is reported. It concerns a 61 year-old patient with tophaceous deposits for 30 years. Descriptive clinical and radiographic images are furnished. PMID- 20012866 TI - Multicenter validation of the value of BASFI and BASDAI in Chinese ankylosing spondylitis and undifferentiated spondyloarthropathy patients. AB - The objectives of this study were to evaluate the reliability of Bath ankylosing spondylitis functional index (BASFI) and Bath ankylosing spondylitis disease activity index (BASDAI) in Chinese ankylosing spondylitis (AS) and undifferentiated spondyloarthropathy (USpA) patients. 664 AS patients by the revised New York criteria for AS and 252 USpA patients by the European Spondyloarthropathy Study Group criteria were enrolled. BASDAI and BASFI questionnaires were translated into Chinese. Participants were required to fill in BASFI and BASDAI questionnaires again after 24 h. Moreover, BASDAI and BASFI were compared in AS patients receiving Enbrel or infliximab before and after treatment. For AS group, BASDAI ICC: 0.9502 (95% CI: 0.9330-0.9502, alpha=0.9702), BASFI ICC: 0.9587 (95% CI: 0.9521-0.9645, alpha=0.9789). For USpA group, BASDAI ICC: 0.9530 (95% CI: 0.9402-0.9632, alpha=0.9760), BASFI ICC: 0.9900 (95% CI: 0.9871-0.9922, alpha=0.9950). In the AS group, disease duration, occipital wall distance, modified Schober test, chest expansion, ESR, and CRP showed significant correlation with BASDAI and BASFI (all P<0.01). In the USpA group, onset age, ESR, and CRP were significantly correlated with BASDAI (all P<0.05), while modified Schober test, ESR, and CRP were significantly associated with BASFI (all P<0.05). The change in BASDAI and BASFI via Enbrel or infliximab treatment showed a significant positive correlation (P<0.01). The two instruments have good reliability and reference value regarding the evaluation of patient's condition and anti-TNF-alpha treatment response. PMID- 20012867 TI - Severity of carpal tunnel syndrome assessed with high frequency ultrasonography: reply to Karadag and colleagues. PMID- 20012868 TI - Functional capacity in rheumatoid arthritis patients: comparison between Spanish and Brazilian sample. AB - The main objective of this study is to compare Spanish and Brazilian self reported health-related functional capacity in patients with rheumatoid arthritis (RA). 197 patients diagnosed with RA were studied in Spain (n = 127) and Brazil (n = 70). Pain (Visual analog scale) and functional capacity (Health Assessment Questionnaire/HAQ) were assessed. Patients were questioned about regular exercise practice. Comparisons between groups were performed with Chi-square tests and Student t test. Pearson's correlation coefficient and linear regression models were used to analyze the associations. Brazilian patients were younger (p = 0.013), had worse levels of pain (p = 0.001) and a trend to experience worse functional capacity (p = 0.057) than Spanish ones. Spanish RA patients had higher body mass index (BMI) (p = 0.019) and longer disease duration (p = 0.001). Also, a higher percentage of subjects with RA from the Spanish cohort had been elected to take early retirement when compared with Brazilian patients (p = 0.010). Spanish RA patients had received more drugs than Brazilians (oral corticosteroids p = 0.010, Leflunomide p = 0.023, Methotrexate p = 0.072, non-steroidal anti inflammatory drugs p = 0.064, biologic therapies p = 0.001). The functional capacity (HAQ) was correlated with age (p = 0.001), disease duration (p = 0.001), age at diagnosis (p = 0.001), pain (p = 0.001) and BMI (p = 0.001) in Spanish patients. In Brazilian, these correlations were only found with disease duration (p = 0.004) and pain (p = 0.001). In conclusion, our data suggest a better management of RA in Spanish when compared with Brazilians. Even with less pain and functional capacity, they receive more drug treatment and a higher percentage of them are retired early. PMID- 20012869 TI - Gender differences in the relationship of anti-parvovirus B19 IgG with antinuclear antibody and C-reactive protein in clinical adult serum samples. AB - Human parvovirus B19 (B19) infection is often suspected as an etiologic agent in a variety of rheumatologic diseases. It has been hypothesized that this virus potentially induces immune dysregulation by abnormal cytokine expression in susceptible hosts. The objective of this study is to examine the relationship between anti-parvovirus B19 IgG antibody (B19 IgG) and two common markers of immune dysregulation-antinuclear antibody (ANA) and C-reactive protein (CRP) in clinical sera. Qualitative B19 IgG antibody and levels of high-sensitivity CRP were determined in adult serum samples submitted to a university hospital clinical laboratory for ANA testing. Prevalence of B19 IgG was compared among groups by ANA status and CRP tertile. B19 IgG was detected in 72.3% of 318 samples. Among women above the first quartile of age (>38 years), presence of B19 IgG was associated with CRP tertile rank (P = 0.008) and CRP levels > or =1 mg/L (P = 0.001) independent of age and ANA status. B19 IgG was less frequent in ANA positive than ANA-negative women < or =38 years of age (P = 0.009). Viral antibody was not associated with either biomarker in men. These data suggest parvovirus B19 infection may be associated with chronic inflammation in some women after the third decade of life. PMID- 20012870 TI - QT interval dispersion in North Indian children with Kawasaki disease without overt coronary artery abnormalities. AB - Increased QT interval dispersion has been associated with an increased risk for ventricular arrhythmias and sudden cardiac events. We examined the QT interval dispersion in 20 North Indian children with Kawasaki disease (KD) with no coronary artery abnormalities on echocardiography compared the same with matched controls. The study population consisted of 20 children in convalescent phase of KD and 20 age and sex-matched healthy controls. Intervals were measured with the use of a digital caliper with least count of 0.01 mm by a single blinded observer. The QTc dispersion was calculated as the difference between the maximum and minimum corrected QT intervals in 12 and 8 leads (i.e. the 6 precordial leads, the shortest extremity lead, and the median of the 5 other extremity leads). Of the 480 leads obtained (12 per subject), 36 were excluded from analysis (15 because of poor T wave formation and 11 because of presence of U waves). Children with KD had significantly higher QTc dispersion in 12 lead (67.08 +/- 17.72 ms compared to 47.63 +/- 16.48 ms in controls P <= 0.001) as well as 8 lead (60.51 +/- 18.54 ms compared to 42.92 +/- 18.03 ms in controls P <= 0.001) analysis. There was no correlation between delay in IVIG therapy and QT interval dispersion. In conclusion, QT interval dispersion is significantly increased in North Indian children with KD. The dispersion is indicative of inhomogenous ventricular repolarization and may represent increased risk for developing ventricular arrhythmia in this population. PMID- 20012871 TI - A single-nucleotide polymorphism of the TNFSF4 gene is associated with systemic lupus erythematosus in Chinese Han population. AB - Systemic lupus erythematosus (SLE) is a prototypic autoimmune disease with complex genetic inheritance. Recently, single-nucleotide polymorphisms (SNPs) in tumor necrosis factor (TNF) superfamily gene TNFSF4 have been shown to be associated with SLE in European and Hong Kong Chinese populations. But it is unknown whether it is also associated with the disease in Mainland Chinese Han population. We genotyped the SNPs rs1234315 near the TNFSF4 gene in 1,344 SLE patients and 4,315 controls of Chinese Han population and confirmed the association between the SNP and the SLE [odds ratios (ORs) of 1.45 and P values of 1.5 * 10(-16)]. The stratification analyses showed that rs1234315 was more strongly associated with SLE patients with arthritis. Our study not only suggested that the TNFSF4 gene was associated with SLE in Chinese Han population, but also implied that it might be a common genetic factor predisposing to the development of SLE in multiple populations. PMID- 20012872 TI - Effects of a protocol of ischemic postconditioning and/or captopril in hearts of normotensive and hypertensive rats. AB - Brief periods (a few seconds) of cyclic coronary occlusions applied early in reperfusion induce a cardioprotection against infarct size, called postconditioning (PostC) in which B(2)-bradykinin receptors play a pivotal role. Since angiotensin-converting enzyme (ACE) inhibitors reduce degradation of kinins, we studied the effects of PostC on infarct size and postischemic myocardial dysfunction in both normotensive (WKY) and spontaneously hypertensive rats (SHR) acutely or chronically treated with the ACE inhibitor Captopril. Isolated hearts from SHR and WKY rats were subjected to the following protocols: (a) ischemia for 30- and 120-min reperfusion (I/R); (b) I/R + PostC protocol (5 cycles 10-s I/R); (c) pretreatment with Captopril for 4-weeks before to subject the hearts to I/R with or without PostC maneuvers. Some SHR hearts were treated with Captopril during the 20- or 40-min early reperfusion with or without PostC maneuvers. Cardiac function was assessed in vivo with echocardiography. Left ventricular pressure and infarct size were measured ex vivo. Chronic Captopril significantly reduced left ventricular hypertrophy in SHR, and reduced infarct size in both WKY and SHR hearts. PostC maneuvers significantly reduced infarct size in WKY, but not in SHR hearts. Yet, PostC slightly improved postischemic systolic function in untreated SHR. Captopril given in reperfusion was unable to limit I/R injury in SHR hearts. Data show that PostC protection against infarct size is blunted in SHR and that PostC is unable to add its protective effect to those of chronic Captopril, which per se reduces cardiac hypertrophy and heart susceptibility to I/R insult. PMID- 20012873 TI - Microglia: biology and pathology. AB - The past 20 years have seen a gain in knowledge on microglia biology and microglia functions in disease that exceeds the expectations formulated when the microglia "immune network" was introduced. More than 10,000 articles have been published during this time. Important new research avenues of clinical importance have opened up such as the role of microglia in pain and in brain tumors. New controversies have also emerged such as the question of whether microglia are active or reactive players in neurodegenerative disease conditions, or whether they may be victims themselves. Premature commercial interests may be responsible for some of the confusion that currently surrounds microglia in both the Alzheimer and Parkinson's disease research fields. A critical review of the literature shows that the concept of "(micro)glial inflammation" is still open to interpretation, despite a prevailing slant towards a negative meaning. Perhaps the most exciting foreseeable development concerns research on the role of microglia in synaptic plasticity, which is expected to yield an answer to the question whether microglia are the brain's electricians. This review provides an analysis of the latest developments in the microglia field. PMID- 20012874 TI - Relationship between urokinase plasminogen activator receptor (uPAR) and the invasion of human prenatal hair follicle. AB - During the morphogenesis of hair follicles, the invasive migration of basal keratinocytes resembles cell's dissemination of tissue remodeling. The urokinase type plasminogen activator receptor (uPAR) appears to be a key molecule in the metastasis. In order to elucidate the relationship between uPAR and the invasion of the human hair follicle, immunohistochemistry, RT-PCR, plasmids transfection, and western blot were used. The results showed that uPAR was expressed in the outermost epithelial cells of the hair follicle and the basal keratinocytes of epidermis, and that the expression decreased with the development of the hair follicle. The cells of the outer root sheath (ORS) and interfollicle epidermis, which overexpressed uPAR, acquired increased invasiveness; however, they showed decreased invasion with overexpression of the urokinase-type plasminogen activator amino terminal fragment (uPA ATF), which inhibited the combination of uPAR and uPA competitively, and the cell invasive migration with overexpressed uPAR was required activated extracellular signal-regulated kinases (ERK). These results implied that overexpression of uPAR promote the invasive migration of hair follicle into the dermis in uPA-dependent and independent manner during human prenatal development. PMID- 20012876 TI - Complete genome sequence of a tobacco isolate of the tobacco vein banding mosaic virus strain prevailing in China. AB - Tobacco vein banding mosaic virus (TVBMV) is a species of the largest plant virus genus Potyvirus. Its incidence has been increasing in Chinese tobacco-growing area. TVBMV isolates can be clustered into three genetic groups that are corresponding with their geographical origin. We have reported the complete genomic sequence of TVBMV isolate YND with unique NIb/CP cleavage site. Here, we determined and analyzed the complete genomic sequence of isolate HN39, which was collected from tobacco in Henan Province and represented Chinese prevalent strain of TVBMV. HN39 has similar host range with YND, but induce mild vein banding symptom in Nicotiana tabacum cv. Samsun. The genome of TVBMV-HN39 is composed of 9,570 nucleotides, excluding the poly(A) tail. It contains a large ORF of 9,240 nucleotides and encode a polyprotein of 3,079 amino acids. The putative NIa-Pro cleavage site for NIb/CP is Q/G. The identities between the complete genomes of isolates HN39 and YND were 90.0% at nucleotide level and 95.4% at amino acid level. As for other potyviruses, HN39 shared the highest identity with wild tomato mosaic virus (WTMV) at complete genomic level, while different genes shared the highest identities with different potyviruses. This is the second complete genomic sequence of TVBMV reported. PMID- 20012875 TI - Molecular mechanisms underlying the activation of eNOS. AB - Endothelial cells situated at the interface between blood and the vessel wall play a crucial role in controlling vascular tone and homeostasis, particularly in determining the expression of pro- and anti-atherosclerotic genes. Many of these effects are mediated by changes in the generation and release of the vasodilator nitric oxide (NO) in response to hemodynamic stimuli exerted on the luminal surface of endothelial cells by the streaming blood (shear stress) and the cyclic strain of the vascular wall. The endothelial NO synthase (eNOS) is activated in response to fluid shear stress and numerous agonists via cellular events such as; increased intracellular Ca(2+), interaction with substrate and co-factors, as well as adaptor and regulatory proteins, protein phosphorylation, and through shuttling between distinct sub-cellular domains. Dysregulation of these processes leads to attenuated eNOS activity and reduced NO output which is a characteristic feature of numerous patho-physiological disorders such as diabetes and atherosclerosis. This review summarizes some of the recent findings relating to the molecular events regulating eNOS activity. PMID- 20012877 TI - Characteristics of young children with persistent conduct problems 1 year after treatment with the Incredible Years program. AB - In the present study, predictors of persistent conduct problems among children aged 4-8 years were investigated in a randomized controlled trial 1 year after treatment with the Incredible Years parent training program (PT), or combined parent training and child treatment (PT + CT). Data were collected before and after treatment and at a 1-year follow-up. Pre-treatment child characteristics predicting persistent conduct problems in the child at the 1-year follow-up were high levels of internalizing and aggression problems as reported by mothers. The only family characteristic predicting persistence of child conduct problems was having contacts with child protection services. Clinicians and researchers need to closely monitor and identify children with conduct problems not responding to parent training programs. These individuals and their families are likely to need further support. PMID- 20012878 TI - Post exposure prophylaxis of tick-borne relapsing fever. AB - Tick-borne relapsing fever (TBRF) is endemic to Israel. Since 2004, the Israel Defence Forces (IDF) has mandated the prophylaxis of tick-bitten subjects with a five-day doxycycline course. We examined the safety and effectiveness of this policy in preventing TBRF. We analyzed the records from January 2004 to January 2007, and identified all reported events of tick bites or TBRF cases. Data were available on 27 events in which 816 soldiers have undergone physical examination following exposure, and seven TBRF cases were recorded in this group-an attack rate of 0.86% compared with the expected rate of 5.34% from previous army data (relative risk [RR] = 0.16). Of those screened, 128 (15.7%) had tick-bite and were intended for prophylaxis, of which four TBRF cases occurred-3.13% attack rate compared with an expected rate of 38.4% in these bitten individuals without prophylaxis (RR = 0.08, number needed to treat = 3). In all cases in which screening and prophylaxis were provided within 48 h of tick bite, complete prevention of TBRF was achieved. No cases of Jarisch-Herxheimer reaction (JHR) was recorded. Tick-bite screening and prophylactic treatment with doxycycline in endemic areas is a practical, safe, and highly effective policy for preventing TBRF. PMID- 20012879 TI - Experimental analyses of synergistic combinations of antibiotics with a recently recognised antibacterial agent, lacidipine. AB - The cardiovascular drug lacidipine (Lc) is known to possess antibacterial activity. Further potentiation of action is possible by synergism between Lc and an antibiotic or a non-antibiotic. The minimum inhibitory concentration (MIC) of antibiotics, Lc and other non-antibiotics were detected by the agar dilution technique in different bacteria. Synergism was determined by disc diffusion assay, the fractional inhibitory concentration (FIC) index through checkerboard assessment and, also, the protective capacity of the combination by administering the drugs along with 50 x LD(50) challenge dose of virulent Salmonella typhimurium in animal experiments. Synergism between Lc and penicillin was found to be statistically significant (P 160 ng/ml) increases biofilm biomass and, conversely, DNase treatment (>0.03 mg/ml) decreases biofilm biomass at later time points of biofilm development. We present evidence for the role of eDNA in C. albicans biofilm structure and formation, consistent with eDNA being a key element of the ECM in mature C. albicans biofilms and playing a predominant role in biofilm structural integrity and maintenance. PMID- 20012896 TI - Human tissues for research purposes: a conference in the House of Lords. AB - The challenges to using human tissues in research are many and varied. However, there is little consensus on how concerns raised by researchers should be addressed, and who should be responsible for ensuring that patients continue to benefit from medical research carried out using human tissues which have been ethically donated or collected after surgery, or where organs donated for transplant are unsuitable for this purpose. A conference in the House of Lords sought to bring together stakeholders from all areas of human tissue research to discuss the problems experienced, share solutions, and form a Working Party to carry the conference momentum forward into action in the near future. PMID- 20012898 TI - Submarine wastewater discharges: dispersion modelling in the Northern Adriatic Sea. AB - BACKGROUND, AIM AND SCOPE: Opposite interests must coexist in coastal areas: the presence of significant cities and urban centres, of touristic and recreational areas, and of extensive shellfish farming. To avoid local pollution caused by treated wastewaters along the Northern Adriatic coast (Friuli Venezia-Giulia and Veneto regions), marine outfall systems have been constructed. In this study, the application of a numerical dispersion model is used to support the traditional monitoring methods in order to link information concerning the hydrodynamic circulation and the microbiological features, to evaluate possible health risks associated with recreational and coastal shellfish farming activities. The study is a preliminary analysis of the environmental impact of wastewater treatment plants (WWTPs) with submarine discharge outfalls. It also could be useful for the water profile definition according to the Directive 2006/7/EC on the quality of bathing water and for the integrated areal analysis (Ostoich et al. 2006), to define the area of influence of each submarine discharge point. MATERIALS AND METHODS: Historical data on discharges of the considered WWTPs were recovered and evaluated. Data on discharges' control for Veneto region (WWTPs of Lido and Cavallino) were produced by the WWTPs' manager Veritas Laboratory service, while data for the WWTPs of Friuli Venezia-Giulia region were produced by the regional environmental protection agency in the institutional control activity following official methods. The hydrodynamic model used in this work is the three dimensional version of the finite element model SHYFEM, developed at ISMAR-CNR (Marine Science Institute of the Italian National Research Council) in Venice (Umgiesser et al. J Mar Syst 51:123-145, 2008). RESULTS AND DISCUSSION: Numerical simulations have been carried out with the 3D version of the finite element model SHYFEM for 3 months during autumn 2007 to evaluate the bacterial pollution dispersion along the coasts of Veneto and Friuli Venezia-Giulia regions, prescribing meteo-marine forcings and concentration values at the points corresponding to the positions of the submarine outfalls. Model results show that during autumn 2007 the discharges of the submarine outfalls of the Venice province seem to have no impact on the surface water quality, while there are some visible effects in the Gulf of Trieste. This reflects the behaviour of the experimental data collected by ARPAV and ARPA FVG and monitoring campaigns both on water and shellfish quality. Further results have been elaborated to identify the area of influence of each discharge point; scenarios were developed with imposed concentrations. The results seem to highlight that the two discharges of the Veneto region are not noticeable, while the discharges of the Gulf of Trieste (in particular the Servola and Barcola ones) are perceptible. CONCLUSIONS: This study represents a new step towards the study of the microbiological pollution dispersion and impact due to the discharges of the submarine outfalls of the Veneto and Friuli Venezia-Giulia regions (nine considered discharge points). With the 3D version of the finite element model SHYFEM, the information obtained from the hydrodynamic circulation has been linked to the classical methods of analysis, to assess possible risks connected to the microbiological parameter Escherichia coli. RECOMMENDATIONS AND PERSPECTIVES: In future studies the time scale for microbiological parameters' decay could be linked to various environmental parameters such as light climate, temperature, and salinity. Interesting information would come from the study of new scenarios with different configurations of the discharge of the pipelines and/or the treatment plants and in particular from the improvements of the 3D version of the SHYFEM model, to take the stratification process into account which occurs during spring-summer, since the Northern Adriatic Sea is a very complex ecosystem, both as physical and ecological processes. PMID- 20012897 TI - Concentration of trace elements in blood and feed of homebred animals in Southern Serbia. AB - BACKGROUND, AIM AND SCOPE: The paper presents concentrations of trace elements in blood of homebred animals (cows and sheep) from Southern Serbia (Bujanovac) and the contents of natural and anthropogenic radionuclides and some heavy metals in feed. The region of Southern Serbia was exposed to contamination by depleted uranium ammunition during NATO attacks in 1999 and therefore, is of great concern to environmental pollution and human and animal health. MATERIALS AND METHODS: Conventional instrumental and epithermal neutron activation analyses are used to measure trace elements in cow and sheep blood samples collected randomly at six locations in the region of Bujanovac (village of Borovac) in the spring of 2005. Samples of feed (grass and crops: corn, wheat and oats), collected on the same locations (households), are analysed for the contents of radionuclides on an HPGe detector (Ortec, relative efficiency 23%) by standard gamma spectrometry. The content of Hg, Pb and Cd in feed is determined by standard atomic absorption spectrometry on the VarianSpectra220/ThermoSolar GFS97 spectrometer. RESULTS: Concentrations of 29 elements (Na, Al (P), Cl, K, Sc, Cr, Mn, Ni, Fe, Co, Zn, Se, As, Br, Sr, Rb, Sb, In, I, Ba, Cs, La, Nd, Eu, Sm, Tb, Hf, Ta and Th) are determined in blood of the examined animals. In feeds, natural (40)K is found in all of the samples, cosmogenic (7)Be and fission product (137)Cs are detected only in the grass samples, while heavy metals Hg, Cd and Pb are found in the range of 0.01-0.02, 0.84-1.15 and 0.74-7.34 mg/kg, respectively. Calculated soil to-blood transfer factors are in a wide range of 8 x 10(-6) to 64, as a result of varying significance of the elements in animal metabolism and feeding habits. DISCUSSION: The results of trace elements' concentrations in animal blood are in good agreement with available data for K, Ni, Zn, Se and Rb. Higher Br concentrations in animal blood are most probably caused by large biomass burning events during blood sampling. Very low concentration of Fe in cows and sheep confirms the results of previous biochemical studies on animal anaemia in the region. High concentration of As correlates with geochemical peculiarities of the Balkans and is also likely influenced by the use of pesticides in the agricultural production. For some of the elements (La, Nd, Eu, Sm, Tb, Sb, Hf, Ta, Th, In, Ba, Sr, Sc and Cs), there are few or no literature data. Therefore, some of the presented data are significant not only for the country and the region, but on a wider scale. Activities of natural radionuclides in feeds are within the average values reported for the region, while the activities of (210)Pb and (235/238)U are below the limit of detection. This is in accordance with previous investigations showing no widespread contamination by depleted uranium in the area. Contents of Hg and Pb in feeds are below the nationally permissible levels, unlike the content of Cd which exceeds it, probably caused by the use of phosphate fertilisers and fossil fuel combustion in the area. CONCLUSIONS: In general, the concentrations of trace elements in blood of homebred cows and sheep are in good agreement with reference materials, available literature data and the results of previous studies in the area. The exceptions are Fe, As and Br. The contents of natural and anthropogenic radionuclides in feeds are within the expected levels, and there are no signs of contamination by depleted uranium or other fission products. Apart from Cd, there are no signs of pollution by heavy metals in feeds. The highly sensitive method of instrumental neutron activation analysis provides data on the concentration of some elements in animal blood not previously reported for the region and elsewhere. RECOMMENDATIONS AND PERSPECTIVES: The presented study is a part of the long term ongoing project on the health risk assessment on animals and humans in the region. The collected data is intended to provide a base for the animal and human risk assessment as well as an estimate of the general pollution status of the environment in the region. Since some of the investigated elements are classified as important trace elements for livestock, the results could also be used to balance and improve the animal diet and thus, improve the growth and reproduction rate. PMID- 20012899 TI - Antioxidative defense and mitochondrial thermogenic response in brown adipose tissue. AB - Cold-exposure activates interscapular brown adipose tissue (IBAT) non-shivering thermogenesis that relies primarily on intensification of metabolic rate and uncoupling. During cold-acclimation, uncoupling in IBAT decreases superoxide (O(2) (.-)) production and as an adaptive response the activities of manganese and copper, zinc superoxide dismutase (Mn- and CuZn-SOD, respectively) are decreased, as well. However, molecular mechanisms governing this SODs adaptive response are still unsolved. Besides, knowing that NO reinforces IBAT uncoupling, we wondered whether nitric oxide (NO) is taking part in SODs regulation? Mn- and CuZn-SOD mRNA and protein expression, uncoupling protein 1 (UCP1), nitrotyrosine and nuclear factor-kappa B (NF-kappaB) immunolabeling, as well as total SOD (tSOD) activity in IBAT of rats subjected to cold (4 +/- 1 degrees C) for 1, 3, 7, 12, 21 and 45 days and treated by l-arginine or N(omega)-nitro-l-arginine methyl ester (l-NAME) were examined. Cold increased UCP1 immunopositivity and decreased tSOD activity during entire cold-acclimation and transiently, (day 3), activated NF-kappaB and increased Mn and CuZn-SOD mRNA expression and nitrotyrosine labeling, suggesting NO involvement in this signaling. However, SODs mRNA expression was decreasing from day 12 till the end of cold-acclimation. l-arginine augmented and prolonged cold-induced UCP1 and nitrotyrosine immunopositivity, NF-kappaB activation and SODs mRNA expression increase, while l NAME expressed an opposite effect. Related to cold, l-arginine decreased, while l NAME increased Mn-SOD protein expression. In contrast, neither low temperature nor both treatments applied affected CuZn-SOD protein expression. The results showed that adaptive decrease in SODs activity on uncoupling-decreased O(2) (.-) production was achieved already at the level of gene transcription and that NO takes part in the regulation of IBAT SOD isoforms. PMID- 20012901 TI - Enhancing the analytical performance of immunoassays that employ metal-enhanced fluorescence. AB - In this work, we used a model assay system (polyclonal human IgG-goat antihuman IgG) to elucidate some of the key factors that influence the analytical performance of bioassays that employ metal-enhanced fluorescence (MEF) using silver nanoparticles (NPs). Cy5 dye was used as the fluorescent label, and results were compared with a standard assay performed in the absence of NPs. Two sizes of silver NPs were prepared with respective diameters of 60 +/- 10 and 149 +/- 16 nm. The absorption spectra of the NPs in solution were fitted accurately using Mie theory, and the dipole resonance of the 149-nm NPs in solution was found to match well with the absorption spectrum of Cy5. Such spectral matching is a key factor in optimizing MEF. NPs were deposited uniformly and reproducibly on polyelectrolyte-coated polystyrene substrates. Compared to the standard assay performed without the aid of NPs, significant improvements in sensitivity and in limit of detection (LOD) were obtained for the assay with the 149-nm NPs. An important observation was that the relative enhancement of fluorescence increased as the concentration of antigen increased. The metal-assisted assay data were analyzed using standard statistical methods and yielded a LOD of 0.086 ng/mL for the spectrally matched NPs compared to a value of 5.67 ng/mL obtained for the same assay in the absence of NPs. This improvement of approximately 66x in LOD demonstrates the potential of metal-enhanced fluorescence for improving the analytical performance of bioassays when care is taken to optimize the key determining parameters. PMID- 20012902 TI - High-resolution investigations of ripple structures formed by femtosecond laser irradiation of silicon. AB - We report on the structural investigation of self-organized periodic microstructures (ripples) generated in Si(100) targets after multishot irradiation by approximately 100-fs to 800-nm laser pulses at intensities near the single shot ablation threshold. Inspection by surface sensitive microscopy, e.g., atomic force microscopy (AFM) or scanning electron microscopy (SEM), and conventional and high-resolution transmission electron microscopy reveal complex structural modifications upon interaction with the laser: even well outside the ablated area, the target surface exhibits fine ripple-like undulations, consisting of alternating crystalline and amorphous silicon. Inside the heavily modified area, amorphous silicon is found only in the valleys but not on the crests which, instead, consist of highly distorted crystalline phases, rich in defects. PMID- 20012900 TI - Is alcohol beneficial or harmful for cardioprotection? AB - While the effects of chronic ethanol consumption on liver have been well studied and documented, its effect on the cardiovascular system is bimodal. Thus, moderate drinking in many population studies is related to lower prevalence of coronary artery disease (CAD). In contrast, heavy drinking correlates with higher prevalence of CAD. In several other studies of cardiovascular mortalities, abstainers and heavy drinkers are at higher risk than light or moderate drinkers. The composite of this disparate relation in several population studies of cardiovascular mortality has been a "U-" or "J-"shaped curve. Apart from its ability to eliminate cholesterol from the intima of the arteries by reverse cholesterol transport, another major mechanism by which HDL may have this cardioprotective property is by virtue of the ability of its component enzyme paraoxonase1 (PON1) to inhibit LDL oxidation and/or inactivate OxLDL. Therefore, PON1 plays a central role in the disposal of OxLDL and thus is antiatherogenic. Furthermore, PON1 is a multifunctional antioxidant enzyme that can also detoxify the homocysteine metabolite, homocysteine thiolactone (HTL), which can pathologically cause protein damage by homocysteinylation of the lysine residues, thereby leading to atherosclerosis. We demonstrated that moderate alcohol up regulates liver PON1 gene expression and serum activity, whereas heavy alcohol consumption had the opposite effects in both animal models and in humans. The increase in PON1 activity in light drinkers was not due to preferential distribution of high PON1 genotype in this group. It is well known that wine consumption in several countries shows a remarkable inverse correlation to local rates of CAD mortality. Significantly, apart from its alcohol content, red wine also has polyphenols such as quercetin and resveratrol that are also known to have cardioprotective effects. We have shown that quercetin also up regulates PON1 gene in rats and in human liver cells. The action of quercetin seems to be mediated via the active form of the nuclear lipogenic transcription factor, sterol-regulatory element-binding protein 2 (SREBP2) that is translocated from endoplasmic reticulum to the nucleus. However, the mechanism of action of ethanol mediated up-regulation of PON1 gene remains to be elucidated. We conclude that both moderate ethanol and quercetin, the two major components of red wine, exhibit cardioprotective properties via the up-regulation of the antiatherogenic gene PON1. PMID- 20012903 TI - Advances in identification of plant gums in cultural heritage by thermally assisted hydrolysis and methylation. AB - Plant gums are present in works of art as binding media for watercolours and adhesives for cellulosic substrates. Thermally assisted hydrolysis and methylation (THM) in combination with analytical pyrolysis coupled to GC/MS has been applied to the characterisation of plant gums typically used in artworks. THM products from standard samples of arabic gum, tragacanth gum and cherry gum were characterised. The main products identified are permethylated and partially methylated aldonic acids, characteristic of specific epimeric sugars. Aldonic acids were formed by alkaline hydrolysis of free reducing sugars and of reducing polysaccharide terminal groups, while methylation occurs during pyrolysis. The presence of these characteristic markers allows gum identification. A systematic analysis of all the parameters that can affect the marker yields was performed. In particular, the influence of pyrolysis temperature, reagent concentration and contact time between tetramethylammonium hydroxide and sample were studied, and different kinds of sample preparation procedures were tested. Some analyses on real watercolours were performed, and gum binders were classified using the peak area ratio of the main monosaccharide markers. PMID- 20012904 TI - Determining the absolute, chemical-heterogeneity-corrected molar mass averages, distribution, and solution conformation of random copolymers. AB - We present a method by which to obtain the absolute, chemical-heterogeneity corrected molar mass (M) averages and distributions of copolymers and apply the method to a gradient random copolymer of styrene and methyl methacrylate in which the styrene percentage decreases from approximately 30% to 19% as a function of increasing molar mass. The method consists of separation by size-exclusion chromatography (SEC) with detection using multi-angle static light scattering (MALS), differential viscometry (VISC), differential refractometry (DRI), and ultraviolet absorption spectroscopy (UV) and relies on the preferential absorption of styrene over methyl methacrylate at 260 nm. Using this quadruple detector SEC/MALS/UV/VISC/DRI approach, the percentage of styrene (%St) in each elution slice is determined. This %St is then used to determine the specific refractive index increment, corrected for chemical composition, at each elution slice, which is then used to obtain the molar mass at each slice, corrected for chemical composition. From this corrected molar mass and from the chemical composition-corrected refractometer response, the absolute, chemical heterogeneity-corrected molar mass averages and distribution of the copolymer are calculated. The corrected molar mass and intrinsic viscosity at each SEC elution slice are used to construct a chemical-heterogeneity-corrected Mark-Houwink plot. The slice-wise-corrected M data are used, in conjunction with the MALS-determined R(G,z ) of each slice, to construct a conformation plot corrected for chemical heterogeneity. The corrected molar mass distribution (MMD) of the gradient copolymer extends over an approximately 30,000 g/mol wider range than the uncorrected MMD. Additionally, correction of the Mark-Houwink and conformation plots for the effects of chemical heterogeneity shows that the copolymer adopts a more compact conformation in solution than originally concluded. PMID- 20012906 TI - Tumor-associated macrophages in clear cell renal cell carcinoma express both gastrin-releasing peptide and its receptor: a possible modulatory role of immune effectors cells. AB - PURPOSE: Renal cell carcinomas (RCC) frequently express the gastrin-releasing peptide receptor (GRP-R). Gastrin-releasing peptide (GRP) stimulates tumor cell proliferation and neoangiogenesis. Tumor-associated macrophages (TAM) comprise an important cellular component of these tumors. We analyzed the GRP/GRP-R network in clear cell RCC (ccRCC) and non-clear cell RCC (non-ccRCC) with special regard to its expression by macrophages, tumor cells and microvessels. METHODS: Gastrin releasing peptide and GRP-R expression in 17 ccRCC and 9 non-ccRCC were analyzed by RT-PCR, immunohistochemistry and double immunofluorescence staining. RESULTS: Tumor-associated macrophages expressed GRP and GRP receptor in ccRCC. Tumor cells and microvessels showed low to intermediate GRP-R expression in nearly all cases. In 12 ccRCC tumor epithelia also expressed low levels of GRP. Microvascular GRP expression was found in nine cases of ccRCC. For non-RCC, the expression of GRP and GRP receptor expression pattern was similar. CONCLUSIONS: Tumor-associated macrophages are the main source of GRP in RCC. GRP receptor on TAM, tumor epithelia and microvessels might be a molecular base of a GRP/GRP receptor network, potentially acting as a paracrine/autocrine modulator of TAM recruitment, tumor growth and neoangiogenesis. PMID- 20012905 TI - IL-17RC: a partner in IL-17 signaling and beyond. AB - The interleukin (IL)-17 cytokine family members IL-17A and IL-17F mediate inflammatory activities via the IL-17 receptor (IL-17R) complex, comprised of the IL-17RA and IL-17RC subunits. Proper regulation of the IL-17 signaling axis results in effective host defense against extracellular pathogens, while aberrant signaling can drive autoimmune pathology. Elucidating the molecular mechanisms underlying IL-17 signal transduction can yield an enhanced understanding of inflammatory immune processes and also create an avenue for therapeutic intervention in the treatment of IL-17-dependent diseases. To date, the fundamental signaling mechanisms used by the IL-17R complex are still incompletely defined. While current structure-function studies have primarily focused on the IL-17RA subunit, recent research indicates that the IL-17RC subunit plays a key role in modulating IL-17 responses. This review will examine what is known regarding IL-17RC function and provide a framework for future work on this subunit and its impact on human health. PMID- 20012907 TI - Defunctioning stoma in high ASA grade, aged patients, with bowel occlusion due to advanced cancer: is it still worthwhile? AB - PURPOSE: The aim of this study was to assess the role of defunctioning stoma (DS) in elderly high-risk patients with bowel obstruction from advanced colorectal cancer, by exploring consistent variables of outcome, because every other procedure was unfeasible. MATERIALS AND METHODS: A 6-year survey in a single surgery unit (between 1999 and 2004) was retrospectively evaluated, allowing to collect a cohort of 75 patients, aged over 65, who overall presented such critical condition. Pre-operatively, American Society of Anaesthesiologist grade classification was used. Post-operative course was monitored by focusing on gauging symptom relief. So, a validated assessment scale was employed to evaluate physical distress symptoms, graduated on a Likert scale and compared at baseline and day 7, on days 7 and 30, post-operatively. Length of hospital stay (LHS), morbidity, in-hospital (within 30 days) and overall mortality (within 6 months) were also assessed. Paired t test was used as statistical analysis to ascertain improvement of symptoms. RESULTS: All symptoms improved significantly (range, p < 0.05 to p < 0.01) within the surveyed time, with exception of vomiting on day 30 (p = 0.14). Average LHS was 22.8 (standard deviation, +/-3.856) days. Overall morbidity was detected in 68 (91%) patients. In-hospital and overall mortality rates accounted for 27 (35.8%) patients and for 48 (100%) patients, respectively. CONCLUSIONS: The role of DS was effective to improve symptom relief but was poor in terms of morbidity and mortality control. So, ethical concerns have to be addressed, and medical treatment or stenting for left-side obstructions only should be considered as alternative procedures. PMID- 20012908 TI - Population-based study of the epidemiology and the risk factors for Pseudomonas aeruginosa bloodstream infection. AB - BACKGROUND: Detailed population-based data on the epidemiology of Pseudomonas aeruginosa bloodstream infections are sparse. We sought to describe the incidence rate, risk factors, and outcomes associated with P. aeruginosa bacteremia in a large Canadian health region. PATIENTS AND METHODS: A retrospective population based surveillance for P. aeruginosa bacteremia was conducted in the Calgary Health Region (CHR, population:approx. 1.2 million) during the period from 2000 to 2006. RESULTS: A total of 284 incident cases of P. aeruginosa bacteremia were identified in CHR residents, corresponding to an annual incidence rate of 3.6/100,000.Nosocomial acquisition accounted for 45% of cases,healthcare associated community onset for 34% of cases,and community-acquired (CA) cases for 21%. Relative to the general population, risk factors for blood stream infection included male sex, increasing age, hemodialysis,solid organ transplant, diagnosis of cancer, heart disease, HIV infection, diabetes mellitus, and/or chronic obstructive airway disease (COPD). Overall mortality was 29%. Factors associated with mortality in univariate analysis included pulmonary focus of infection and co-morbidities, including chronic liver disease, substance abuse, heart disease, COPD, and cancer, and increased with the burden of co-morbidities. Despite those patients with CA disease having fewer co-morbidities,they had a significantly higher mortality rate than either healthcare-associated cases or nosocomial cases(RR 1.88, p = 0.05). CONCLUSIONS: This study documents that P. aeruginosa bacteremic disease is responsible for a significant burden of illness in general populations and identifies those groups at increased risk of infection and subsequent mortality. This information can be used to identify those individuals likely to benefit from empiric anti-pseudomonal therapies. PMID- 20012909 TI - Activation of Janus kinase/signal transducer and activator of transcription 3 pathway by growth hormone-releasing hormone. AB - Growth hormone-releasing hormone (GHRH) can act as a potent growth factor in various cancers. The mitogenic activity of this neuropeptide is exerted through binding to the pituitary type receptors (GHRH-R) or their splice variants (SV). In the present work, we studied whether this hormone can activate the JAK2/STAT3 pathway which plays a crucial role in cancer cell proliferation and is also linked to carcinogenesis. We transfected HeLa human cervical cancer cells, which are not sensitive to GHRH analogs with the pGHRH-R. Transfected cells responded to the GHRH or its antagonist with an increase or a decrease in proliferation, respectively. These results were confirmed by the expression of proliferating cell nuclear antigen. We then showed that these effects are linked to the activation of the JAK2/STAT3 pathway. Our work demonstrates the activation of JAK/STAT3 pathway by GHRH and sheds further light to the mechanisms of the antitumorogenic action of GHRH antagonists. PMID- 20012910 TI - Patterns of failure after stereotactic radiotherapy of intracranial meningioma. AB - The aim of this work is to evaluate patterns of failure in patients with recurrent meningioma after stereotactic radiotherapy. Of 411 patients with intracranial meningioma treated with radiotherapy at our institution, 22 patients with local tumor progression diagnosed by magnetic resonance imaging (MRI) after radiotherapy (RT) were identified and further investigated. The histologic grade of the meningiomas was World Health Organization (WHO) grade I in 54.5%, WHO grade II in 27.3%, and WHO grade III in 9.1% of cases. Fourteen patients had received fractionated stereotactic RT; five patients underwent intensity modulated RT. The median total dose was 57.6 Gy at 1.8 Gy/fraction, five times weekly. Local recurrences were divided into the dosimetric categories "central" ("in-field") and "marginal" ("out-field"). Median follow-up was 59.5 months. Eleven local failures were found to be central, and 11 were marginal. Recurrence free survival (P < 0.05) and site of local recurrence (P < 0.05) depended statistically significantly on histology. Median recurrence-free survival was 46 months for patients with benign meningioma (WHO grade I) and 31.5 months for patients with higher-grade meningioma (WHO grade II/III). In the WHO grade I group, three recurrences were central and nine were marginal, whereas in the WHO grade II/III group seven recurrences were central and one was marginal. Median time to local tumor progression and site of local recurrence significantly depended on histological grade of meningioma. Regarding site of failure, improvement of dose coverage for benign meningiomas and dose escalation for high grade tumors might further improve therapy outcome. PMID- 20012911 TI - Prognostic value of immunohistochemical profile and response to high-dose methotrexate therapy in primary CNS lymphoma. AB - Several biomarkers have been identified as prognostic factors in primary central nervous system lymphoma (PCNSL). However, the correlation between the histogenetic origin of PCNSL and the response to therapy is still unclear. To elucidate the utility of immunophenotypic markers in predicting clinical outcomes, we investigated 27 immunocompetent patients with PCNSL treated with high-dose methotrexate therapy. Of the 27 patients, 25 received whole-brain radiotherapy after high-dose methotrexate. Immunostaining for CD5, CD10, BCL-6, and MUM-1 was used to determine the immunophenotypic profile of diffuse large B cell lymphoma of PCNSL. We then evaluated whether immunophenotypic markers were associated with the response to therapy or patients' survival. The response to induction high-dose methotrexate therapy was determined by magnetic resonance imaging after three courses of i.v. high-dose methotrexate. We categorized B-cell lymphomas into three known subtypes: germinal center B-cell (GCB), activated-GCB, and post-GCB subtypes according to immunohistochemical profile. All the BCL-6 positive samples were co-positive for MUM-1 and therefore classified into activated-GCB subtype. BCL-6 expression in this study was associated with poor progression-free survival (P = 0.038). No immunophenotypic markers or subtypes had a significant effect on the response to high-dose methotrexate therapy. However, the response itself was a significant predictor for both progression free survival (P < 0.001) and overall survival (P = 0.003). Further investigation is needed to assess BCL-6 as a potential prognostic factor in PCNSL. PMID- 20012912 TI - Selective medicated (saline + natural surfactant) bronchoalveolar lavage in unilateral lung contusion. A clinical randomized controlled trial. AB - OBJECTIVE: Open lung and low tidal volume ventilation appear to be a promising ventilation for chest trauma as it can reduce ARDS and improve outcome. Local therapy (e.g. BAL) can be synergic to remove from the lung the debris, mitigate inflammatory cascade and avoid damage spreading to not compromised lung areas. MATERIALS AND METHODS: 44 pulmonary contused patients were randomized to receive broncho-suction and volume controlled low tidal volume ventilation-VCLTVV (Control Group) or the same ventilation plus medicated (saline + surfactant) BAL (Treatment Group). Tidal volume <10 ml/kg, PEEP of 10-12 cm H(2)O and PaO(2) 60 100 mm Hg and PaCO(2) 35-45 mm Hg were used in both groups. BAL was performed using a fiberscope. 4 boluses of 25 ml saline with 2.4 mg/ml of surfactant were introduced into each contused lobe in which, subsequently, 240 mg of surfactant was instilled. RESULTS: All patients survived. In the Control Group 18 patients developed pneumonia, 5 ARDS and days of intubation were 11.50 (3.83) compared to 5.05 (1.21) of Treatment Group in which OI and PaO(2)/FiO(2) significantly improved from 36 h. CONCLUSIONS: VCLTVV alone was not able to prevent ARDS and infection in the Control Group as the reduction of intubation. In the Treatment Group, VCLTVV and medicated BAL facilitated the removal of degradated lung material and recruited the contused lung regions, enabling the healing of the lung pathology. PMID- 20012915 TI - A novel serine metallokeratinase from a newly isolated Bacillus pumilus A1 grown on chicken feather meal: biochemical and molecular characterization. AB - A keratinolytic enzyme (KerA1) secreted by a newly isolated Bacillus pumilus strain A1 cultivated in medium containing chicken feather meal was purified and characterized, and the gene was isolated and sequenced. The molecular mass of the purified enzyme was estimated to be 34,000 Da by sodium dodecyl sulfate (SDS) polyacrylamide gel electrophoresis and gel filtration. The optimum pH and temperature for the purified keratinase were 9.0 and 60 degrees C, respectively, using keratin as a substrate. KerA1 showed a high stability towards nonionic surfactants. It was found to be relatively stable toward the strong anionic surfactant (SDS). The deduced amino acid sequence of the keratinase KerA1 differs from both the organic solvent tolerant protease of B. pumilus 115b and the dehairing protease of B. pumilus UN-31-C-42 by one and nine amino acids, respectively. These results suggest that this keratinase may be a useful alternative and ecofriendly route for handling the abundant amount of waste feathers and for applications in detergent formulations. PMID- 20012914 TI - The endocrine tumor summit 2008: appraising therapeutic approaches for acromegaly and carcinoid syndrome. AB - The Endocrine Tumor Summit convened in December 2008 to address 6 statements prepared by panel members that reflect important questions in the treatment of acromegaly and carcinoid syndrome. Data pertinent to each of the statements were identified through review of pertinent literature by one of the 9-member panel, enabling a critical evaluation of the statements and the evidence supporting or refuting them. Three statements addressed the validity of serum growth hormone (GH) and insulin-like growth factor-I (IGF-I) concentrations as indicators or predictors of disease in acromegaly. Statements regarding the effects of preoperative somatostatin analog use on pituitary surgical outcomes, their effects on hormone and symptom control in carcinoid syndrome, and the efficacy of extended dosing intervals were reviewed. Panel opinions, based on the level of available scientific evidence, were polled. Finally, their views were compared with those of surveyed community-based endocrinologists and neurosurgeons. PMID- 20012916 TI - Proper definitions of MRI contrast enhancement in liver tumors. PMID- 20012917 TI - Pancreatic acinar metaplasia in the distal oesophagus and the gastric cardia: prevalence, predictors and relation to GORD. AB - PURPOSE: The nature of pancreatic acinar metaplasia (PAM) in the gastro oesophageal junction (GOJ) remains obscure. We aimed to estimate its prevalence and investigate into its risk factors in a population-based series of first-time endoscopy patients. METHODS: We investigated consecutive patients, endoscoped for the first time, representing defined catchment area populations. Biopsies were taken immediately below the GOJ and from the distal oesophagus. Endoscopy room based cross-sectional clinical data were supplemented with exposure data from 160 population controls. Associations, expressed as odds ratios (OR), were modelled with multivariable logistic regression. A subsample of 26 patients underwent oesophageal pH monitoring. RESULTS: Among 644 patients (mean age 53 years, 43% men), PAM was found in 121 patients (19%), exclusively above the GOJ in 40 (6%), below GOJ in 67 (10%), and both above and below GOJ in 14 (2%). PAM exclusively above the GOJ and PAM exclusively below the GOJ were both borderline associated with age (2% increase in prevalence per year). PAM exclusively above the GOJ was significantly associated with female gender (OR 2.8, 95% CI 1.3-6.3) and presence of Helicobacter pylori immediately below the GOJ (OR 2.6, 95% CI 1.3-5.4). Out of 21 patients with Barrett's oesophagus (BO), 8 (38%) had PAM above the GOJ. The mean value for percentage time with oesophageal pH < 4.0 was 7.3% (95% CI 4.3 10.2%) among patients who had PAM above the GOJ (reference value 3.4%). CONCLUSIONS: Pancreatic acinar metaplasia might be an age-dependent lesion, associated with H. pylori, female gender and gastro-oesophageal reflux if located above the GOJ. PMID- 20012919 TI - Transarterial renal embolization for enlarged horseshoe kidney. AB - A 56-year-old Japanese man was admitted to our hospital with abdominal fullness in June 2006. He had been diagnosed as having a horseshoe kidney by computed tomography in January 2002. At that time, renal dysfunction (serum creatine: 2.0 mg/dl) was detected. Even after hemodialysis was started for end-stage renal failure in March 2006, his abdominal fullness became progressively worse. CT scanning showed a markedly enlarged horseshoe kidney. Transarterial embolization (TAE) was performed via the right renal arteries with 14 platinum microcoils; the left renal arteries were not embolized in order to preserve sufficient parenchyma and a urine volume of more than 1,000 ml daily. Two years after TAE, a decrease in the size of the left kidney was noted along with the right kidney. However, urine output was still more than 1,000 ml daily. It is possible that one kidney compressed the contralateral kidney, resulting in enlargement of both components of the horseshoe kidney and renal dysfunction. TAE may be a useful option for obstructive uropathy in patients with horseshoe kidney, which has conventionally been treated surgically. PMID- 20012918 TI - The effects of weekly alendronate therapy in Taiwanese males with osteoporosis. AB - The aim of this study was to evaluate the efficacy, safety, and tolerability of weekly alendronate administration on male osteoporosis in Taiwan. This 6-month, randomized, open-label controlled trial enrolled 46 men with osteoporosis who were randomized to either 70 mg alendronate once weekly (n = 23) or control (n = 23). Bone mineral density (BMD) of lumbar spine and hip and biochemical bone turnover markers were measured; adverse events and tolerability were assessed. Subjects treated with alendronate showed a significant increase in BMD of 5.5% (vs. 2% in control group) at the lumbar spine and 2.7% (vs. 0.7%) at the femoral neck (P < 0.05) at 6 months, respectively. There were also significant decreases in serum level of bone formation marker (bone-specific alkaline phosphatase) and urinary excretion of bone resorption marker (deoxypyridinoline) at 3 and 6 months. Thus, alendronate showed anti-osteoporotic effects by increasing BMD and decreasing the concentrations of bone markers. The adverse events were mild and showed no significant difference between the two groups on safety assessments. PMID- 20012920 TI - High-resolution CT of nontuberculous mycobacteria pulmonary infection in immunocompetent, non-HIV-positive patients. AB - PURPOSE: The objective of this study was to demonstrate that nontuberculous mycobacteria (NTM) pulmonary infections are not so infrequent and that the diagnosis may be suggested on the basis of the high-resolution computed tomography (HRCT) pattern alone. MATERIALS AND METHODS: We retrospectively reviewed HRCT scans of 29 patients (9 men, 18 women; mean age 63 years, range 38 88 years) with positive culture from bronchial wash. Mycobacterium avium complex (MAC) was present in all (with the exception of one in whom the NTM was indistinct). In six patients, MAC was associated with M. chelonae, M. kansasii, M. fortuitum or M. xenopi. In one of these patients, MAC was associated with both M. fortuitum and M. chelonae. All patients had had nonspecific symptoms of pulmonary infection for a time ranging from 6 months to 12 years. Previous tuberculous infection was present in five patients (18.5%). Eleven patients had other pulmonary diseases (40.8%), and 12 had associated systemic diseases (44.4%). RESULTS: HRCT findings were apical fibrotic scarring (n=8; 29.6%), consolidations (n=16; 59.2%), single/multiple nodules >1 cm (n=8, multiple; 29.6%), cavitations (n=7; 25.9%), ground glass appearance (n=3; 11.1%), reticular/reticulonodular pattern (n=6; 22.2%), bronchiectasis (n=25; 92.5%), centrilobular nodules (tree in bud) (n=24; 88.8%), air trapping (n=8; 29.6%), lymphadenopathy >1 cm, also with calcification (n=13, 3 with calcification; 48.1%) and pleural effusion (n=2; 7.4%). In 3/7 patients with nodules >1 cm and with cavitations, the "feeding bronchus sign" (a patent bronchus running into a cavitation) was present. Lesions were in the upper lobes in 23 (85.1%), middle lobe/lingula in 25 (92.5%) and lower lobes in 18 (66.6%) patients. The findings were diffuse in 13 (48.1%) cases and patchy in 17 (62.9%). CONCLUSIONS: HRCT findings are essential for the diagnosis of NTM pulmonary infection. The presence of bronchiectasis, cavitary nodules with feeding bronchus sign and tree-in-bud nodules in the middle lobe and lingula are suggestive of NTM infection, thus assisting the physician in the diagnostic workup of these patients. PMID- 20012921 TI - Dose reduction in multislice CT by means of bismuth shields: results of in vivo measurements and computed evaluation. AB - PURPOSE: We investigated the amount of patient dose reduction in the thyroid, lens of the eye and the breast when using bismuth protections in multislice computed tomography (CT) exams as well as their influence on the quality of diagnostic images. MATERIALS AND METHODS: The radiation dose was measured by using thermoluminescence dosimeters. The study was conducted on the two CT scanners installed in our radiology department (64 and eight slices). The shield effects on the CT image were evaluated by measuring the signal-to-noise ratio in a phantom and in vivo, and by verifying the presence of artefacts on patients' images. The obtained organ-dose reduction factors were used to evaluate the effects of shielding on the effective dose. RESULTS: The shielding attenuation ranged from 30% to 60% depending on the CT scan protocols and organs. The difference between shielded and unshielded signal-to-noise ratio was statistically significant but within the standard requirements for quality assurance. Results were in agreement with the radiologists' perception of image quality. The use of the shields allowed up to 38% reduction of effective dose. CONCLUSIONS: Use of bismuth shields significantly decreases both organ and effective radiation dose, with a consequent reduction in health risk for the patient, quantified in 1.4 fewer cases of radiation-induced tumours every 5 years in our centre (12,100 exams/year), in agreement with the risk factors proposed by Publication 60 of the International Commission on Radiological Protection (ICRP). The relative inexpensiveness of these protections, their easy application and their substantial lack of influence on image quality suggest their massive introduction into routine clinical practice. PMID- 20012922 TI - Diagnostic imaging in the study of human hepatobiliary fascioliasis. AB - PURPOSE: Fascioliasis is a rare zoonotic disease caused by the trematode Fasciola hepatica. We present the typical patterns of hepatobiliary fascioliasis observed in ten patients studied with multimodality imaging. MATERIALS AND METHODS: Between 2002 and 2005, ten women with fascioliasis were admitted to the Brigham and Women's Hospital, Harvard Medical School (BWH), with abdominal pain and mild fever. All imaging modalities, including ultrasound (US), computed tomography (CT), magnetic resonance (MR) imaging (n = 2) and endoscopic retrograde cholangiopancreatography (ERCP) (n = 1) were reviewed by two expert radiologists working in consensus. RESULTS: In all patients (10/10, 100%), US showed parenchymal heterogeneity characterised by multiple subcapsular and peribiliary hypoechoic nodular lesions that were ill-defined and coalesced into tubular or tortuous structures. In six patients (6/10, 60%), the lesions appeared hypoechoic, whereas in four patients (4/10, 40%), there was an alternation of hyperechoic and hypoechoic nodules. On CT, all patients (10/10, 100%) showed hypodense patchy lesions in subcapsular, peribiliary or periportal locations, which coalesced to form tubular structures and were more evident during the portal phase. Lesion diameter ranged from 2 cm to 7 cm. Capsular enhancement was seen in four cases on CT (4/10, 40%) and in one also at MR imaging. MR imaging, performed in two patients, confirmed the presence of the lesions, which appeared hyperintense on T2-weighted images and were characterised by mild peripheral enhancement after gadolinium administration. Four patients had gallbladder wall thickening (4/10, 40%), with parasites in the gallbladder lumen. CONCLUSIONS: Although rare, hepatobiliary fascioliasis should be considered in the differential diagnosis in the appropriate clinical scenario, especially in patients coming from endemic areas. The typical imaging pattern of fascioliasis is the presence of subcapsular, peribiliary or periportal nodules that are usually ill-defined and coalesce, giving rise to a tubular or tortuous appearance. PMID- 20012923 TI - Scleroderma renal crisis: a rare but severe complication of systemic sclerosis. AB - Scleroderma renal crisis (SRC) is a major complication in patients with systemic sclerosis (SSc). It is characterized by malignant hypertension and oligo/anuric acute renal failure. SRC occurs in 5% of patients with SSc, particularly in the first years of disease evolution and in the diffuse form. The occurrence of SRC is more common in patients treated with glucocorticoids, the risk increasing with increasing dose. Left ventricular insufficiency and hypertensive encephalopathy are typical clinical features. Thrombotic microangiopathy is detected in 43% of the cases. Anti-RNA-polymerase III antibodies are present in one third of patients who develop SRC. Renal biopsy is not necessary if SRC presents with classical features. However, it can help to define prognosis and guide treatment in atypical forms. The prognosis of SRC has dramatically improved with the introduction of angiotensin-converting enzyme inhibitors (ACEi). However, 5 years survival in SSc patients who develop the full picture of SRC remains low (65%). SRC is often triggered by nephrotoxic drugs and/or intravascular volume depletion. The treatment of SRC relies on aggressive control of blood pressure with ACEi, if needed in combination with other types of antihypertensive drugs. Dialysis is frequently indicated, but can be stopped in approximately half of patients, mainly in those for whom a perfect control of blood pressure is obtained. Patients who need dialysis for more than 2 years qualify for renal transplantation. PMID- 20012925 TI - MicroRNA function in cancer: oncogene or a tumor suppressor? AB - MicroRNAs (miRNAs) are small noncoding, double-stranded RNA molecules that can mediate the expression of target genes with complementary sequences. About 5,300 human genes have been implicated as targets for miRNAs, making them one of the most abundant classes of regulatory genes in humans. MiRNAs recognize their target mRNAs based on sequence complementarity and act on them to cause the inhibition of protein translation by degradation of mRNA. Besides contributing to development and normal function, microRNAs have functions in various human diseases. Given the importance of miRNAs in regulating cellular differentiation and proliferation, it is not surprising that their misregulation is linked to cancer. In cancer, miRNAs function as regulatory molecules, acting as oncogenes or tumor suppressors. Amplification or overexpression of miRNAs can down-regulate tumor suppressors or other genes involved in cell differentiation, thereby contributing to tumor formation by stimulating proliferation, angiogenesis, and invasion; i.e., they act as oncogenes. Similarly, miRNAs can down-regulate different proteins with oncogenic activity; i.e., they act as tumor suppressors. This review will highlight the recent discoveries regarding miRNAs and their importance in cancer. PMID- 20012926 TI - [Problems in microbial safety of advanced therapy medicinal products. Squaring the circle]. AB - Today, sterility of parenteral drugs is practically guaranteed. Well-defined procedures in the pharmaceutical industry enable effective protection against contamination by bacteria and fungi. In contrast, problems regarding microbial safety of advanced therapy medicinal products (ATMPs), especially of cell therapeutics, are at best only partially solved. The latter should be understood as a challenge for manufacturers, regulators, and physicians. Many of the manufacturing principles mentioned above are not applicable in production of cell therapeutics. Sterility of source materials cannot be guaranteed and the hitherto known procedures for sterilization are, as a rule, not feasible. Thus, the sterility of the final product cannot be guaranteed. Considering the extremely short shelf life of many cell therapeutics, sometimes only a few hours, the results from established methods for sterility testing are often available too late. Furthermore, the sterility of a test sample does not indicate sterility of the whole product. In most cases, conventional methods for pyrogen testing are not applicable for ATMPs. This paper demonstrates relevant limitations regarding microbial safety and pyrogenicity. Possibilities to overcome these problems are discussed and some novel solutions are proposed. PMID- 20012924 TI - Role of epithelial-to-mesenchymal transition (EMT) in drug sensitivity and metastasis in bladder cancer. AB - Epithelial-to-mesenchymal transition (EMT) is a process that plays essential roles in development and wound healing that is characterized by loss of homotypic adhesion and cell polarity and increased invasion and migration. At the molecular level, EMT is characterized by loss of E-cadherin and increased expression of several transcriptional repressors of E-cadherin expression (Zeb-1, Zeb-2, Twist, Snail, and Slug). Early work established that loss of E-cadherin and increased expression of MMP-9 was associated with a poor clinical outcome in patients with urothelial tumors, suggesting that EMT might also be associated with bladder cancer progression and metastasis. More recently, we have used global gene expression profiling to characterize the molecular heterogeneity in human urothelial cancer cell lines (n = 20) and primary patient tumors, and unsupervised clustering analyses revealed that the cells naturally segregate into two discrete "epithelial" and "mesenchymal" subsets, the latter consisting entirely of muscle-invasive tumors. Importantly, sensitivity to inhibitors of the epidermal growth factor receptor (EGFR) or type-3 fibroblast growth factor receptor (FGFR3) was confined to the "epithelial" subset, and sensitivity to EGFR inhibitors could be reestablished by micro-RNA-mediated molecular reversal of EMT. The results suggest that EMT coordinately regulates drug resistance and muscle invasion/metastasis in urothelial cancer and is a dominant feature of overall cancer biology. PMID- 20012927 TI - Regulatory requirements for clinical trial and marketing authorisation application for gene therapy medicinal products. AB - Over the last two decades, clinical trials using gene therapy medicinal products (GTMPs) have been carried out for a large number of rare, inherited monogeneic disorders as well as common multigeneic diseases such as cancer, cardiovascular and infectious diseases including AIDS. Despite some early difficulties and setbacks, the gene therapy field has slowly progressed and, nowadays, offers the promise of novel treatments for a growing number of diseases. On the other hand, gene therapy approaches are often associated with additional risks due to limited clinical experience with a given gene transfer system, long-lasting effects of the therapeutic gene, and/or a complex mode of action. As a result, specific regulations and guidelines have been introduced within the EU to help address these uncertainties. This article summarises the legislative framework and will provide an overview on the regulatory requirements for clinical trials and marketing authorisation applications. PMID- 20012928 TI - [Applications for medical rehabilitation in the field of dermatology]. AB - New rehabilitation guidelines and laws have been introduced to promote competition. As a result, the application procedures for rehabilitation measures have been reformed. Since 2007, the application procedure depends on the type of benefit (e.g. secondary or tertiary prevention) and the insurance provider (statutory health or pension insurance company). Thus in order to prescribe rehabilitation as a form of tertiary prevention payable by a statutory health insurance company, accredited doctors have to demonstrate that they have the necessary qualifications as stipulated by the rehabilitation guidelines. As before 2007, any accredited doctor may however apply for rehabilitation in the sense of tertiary prevention payable by a statutory pension insurance company, and for benefits associated with primary and secondary prevention payable by a statutory health insurance company, without providing corresponding proof of qualification. In addition, dermatologists should report occupational skin diseases to the statutory accident insurance using the "optimized dermatologist's report" which also allows them to recommend secondary and tertiary preventative measures. Every insurance-accredited dermatologist should understand the application process and the contents of preventative and rehabilitation measures to ensure their patients' right to participate. PMID- 20012929 TI - [Acne inversa]. AB - Acne inversa is a chronic inflammatory disease of the hair follicles. It can lead to severe functional and psychological impairment. It is characterized by inflamed painful nodules, abscesses, fistulas and scarring in late stages of the disease. The causes of acne inversa are still not fully understood. Conservative treatment options such as antibiotics may lead to clinical improvement; however they do not produce healing. Therapy of choice, especially in severe forms, is radical wide excision of all affected areas. Despite a variety of treatment options, acne inversa is still a therapeutic challenge. PMID- 20012930 TI - [Liver fibrosis - pathogenesis and novel therapeutic approaches]. AB - Chronic liver injury, such as viral hepatitis, alcohol, metabolic syndrome or other toxic damages, leads to an inflammatory response including the infiltration and activation of immune cells and to the proliferation and transdifferentiation of mesenchymal cells within the liver, especially of hepatic stellate cells. These cells produce an excess of extracellular matrix proteins that are deposited in the liver. Hepatic fibrosis may progress to liver cirrhosis and liver failure. This review aims at summarizing the current view on the pathogenic sequence during fibrogenesis highlighting the essential role of cytokines and chemokines. Understanding the complex cellular interactions in liver fibrosis may help to develop novel antifibrotic therapies in the near future. PMID- 20012931 TI - Mating flights select for symmetry in honeybee drones (Apis mellifera). AB - Males of the honeybee (Apis mellifera) fly to specific drone congregation areas (DCAs), which virgin queens visit in order to mate. From the thousands of drones that are reared in a single colony, only very few succeed in copulating with a queen, and therefore, a strong selection is expected to act on adult drones during their mating flights. In consequence, the gathering of drones at DCAs may serve as an indirect mate selection mechanism, assuring that queens only mate with those individuals having a better flight ability and a higher responsiveness to the queen's visual and chemical cues. Here, we tested this idea relying on wing fluctuating asymmetry (FA) as a measure of phenotypic quality. By recapturing marked drones at a natural DCA and comparing their size and FA with a control sample of drones collected at their maternal hives, we were able to detect any selection on wing size and wing FA occurring during the mating flights. Although we found no solid evidence for selection on wing size, wing FA was found to be significantly lower in the drones collected at the DCA than in those collected at the hives. Our results demonstrate the action of selection during drone mating flights for the first time, showing that developmental stability can influence the mating ability of honeybee drones. We therefore conclude that selection during honeybee drone mating flights may confer some fitness advantages to the queens. PMID- 20012932 TI - [Disturbances of slow-wave sleep and psychiatric disorders]. AB - Slow-wave sleep is defined as sleep stages 3 and 4 that characteristically show slow delta EEG activity during polysomnography. The percentage of slow-wave sleep normally declines with age. Sleep disorders are a common symptom of many psychiatric disorders. In polysomnographic recordings they mostly manifest as disturbances of sleep continuity. In some disorders changes in REM sleep are also found. A reduction of slow-wave sleep has most often been described in patients with depression and addictive disorders. More recent research implicates slow wave sleep as an important factor in memory consolidation, especially the contents of declarative memory. Psychotropic drugs influence sleep in different ways. Hypnotic substances can reduce the deep sleep stages (e.g. benzodiazepines), whereas 5-HT2C antagonists increase the percentage of slow-wave sleep. Whether a selective impairment/alteration of slow-wave sleep is clinically relevant has not yet been proved. PMID- 20012933 TI - Characterizing variation in mycorrhiza effect among diverse plant varieties. AB - Exploitation of arbuscular mycorrhizal fungi may be an important approach for development of reduced-input agriculture. We discuss the use of linear models to analyze variation in mycorrhiza response among diverse plant varieties in order to assess the value of mycorrhizas. Our approach allows elimination of variation linked to differences in plant performance in the absence of mycorrhizas and the selection of plant lines that might harbor genetic variation of use to improve the mycorrhizal symbiosis in agriculture. We illustrate our approach by applying it to previously published and to novel data. We suggest that in dealing with a relative trait such as mycorrhiza effect, the choice of measure used to quantify the trait greatly affects interpretation. In the plant populations under consideration, we find evidence for a greater potential to increase mycorrhiza benefit than previously suggested. PMID- 20012934 TI - QTL mapping of clubroot resistance in radish (Raphanus sativus L.). AB - A QTL analysis for clubroot resistance (CR) of radish was performed using an F(2) population derived from a crossing of a CR Japanese radish and a clubroot susceptible (CS) Chinese radish. F(3) plants obtained by selfing of F(2) plants were used for the CR tests. The potted seedlings were inoculated and the symptom was evaluated 6 weeks thereafter. The mean disease indexes of the F(3) plants were used for the phenotype of the F(2). The results of two CR tests were analyzed for the presence of QTL. A linkage map was constructed using AFLP and SSR markers; it spanned 554 cM and contained 18 linkage groups. A CR locus was observed in the top region of linkage group 1 in two tests. Therefore, the present results suggest that a large part of radish CR is controlled by a single gene or closely linked genes in this radish population, although minor effects of other genomic areas cannot be ruled out. The CR locus was named Crs1. Markers linked to Crs1 showed sequence homology to the genomic region of the top of chromosome 3 of Arabidopsis, as in the case of Crr3, a CR locus in Brassica rapa. These markers should be useful for breeding CR cultivars of radish. As Japanese radishes are known to be highly resistant or immune to clubroot, these markers may also be useful in the introgression of this CR gene to Brassica crops. PMID- 20012935 TI - Must we always follow the instructions of automated external defibrillators? PMID- 20012936 TI - Injury patterns to the posteromedial corner of the knee in high-grade multiligament knee injuries: a MRI study. AB - Injury patterns to the posteromedial corner of the knee have not been previously studied in the context of multiligament knee injuries. We performed a retrospective magnetic resonance imaging and clinical review of a consecutive series of 27 dislocatable knees presenting to a single level-one trauma center from 2005 to 2008. Post-injury magnetic resonance imaging studies were reviewed by two fellowship-trained musculoskeletal radiologists to assess injury patterns to the posteromedial corner. In our series, injury to at least one structure within the posteromedial corner was observed in 81% (22/27) of cases while injury to the superficial medial collateral ligament alone was seen in 63% (17/27) of cases. Furthermore, injuries to the posterior horn of the medial meniscus were associated with a tear of the meniscotibial ligaments in all cases and with a tear of the posterior oblique ligament in 67% of cases. All patients with grade III laxity (>10 mm medial opening) under an examination under anesthesia had a complete tear of the posterior oblique ligament and meniscotibial ligament in addition to a medial collateral ligament injury. Injury to the semimembranosus attachment alone was not associated with clinically significant laxity under an examination under anesthesia. Our findings demonstrate that injuries to the posteromedial corner are common in the setting of traumatic knee dislocations. Interestingly, high-grade medial instability during an examination under anesthesia and injury to the posterior horn of the medial meniscus may be important indicators for further posteromedial corner injury. PMID- 20012937 TI - Mechanized pivot shift test achieves greater accuracy than manual pivot shift test. AB - The objective of this study was to design a navigated mechanized pivot shift test setup and evaluate its repeatability in the ACL-deficient knee. It was hypothesized that translations and rotations measured with the mechanized pivot shift would be more repeatable when compared to those obtained with a manual pivot shift. Twelve fresh frozen cadaveric hip-to-toe whole lower extremities were used for this study. A manual pivot shift test was performed in the intact knee and in the ACL-deficient knee and was repeated three times. A navigation system simultaneously recorded tibial translation and rotation. The mechanized pivot shift test consists of a modified continuous passive motion (CPM) machine and a custom-made foot holder to allow for the application of internal rotation moments at the knee. Valgus moments were achieved by a 45 degrees tilt of the CPM machine with respect to the supine position and a Velcro strap secured across the proximal tibia. The mechanized pivot shift was repeated three times. Repeated measures ANOVA was used to compare manual and mechanized pivot shift testing. An intra-class correlation coefficient (ICC) was used to determine variability within each knee at each testing condition. In the ACL-deficient knee, translation with manual pivot shift testing (11.7 +/- 2.6 mm) was significantly higher than with mechanized pivot shift testing (7.4 +/- 2.5 mm; p < 0.05). Rotation with the manual pivot shift testing (18.6 +/- 5.4 degrees) was also significantly higher than with mechanized pivot shift testing (11.0 +/- 2.3 degrees; p < 0.05). The intra-class ICC for translations was 0.76 for manual pivot shift and 0.92 for the mechanized pivot shift test. The intra-class ICC for rotations was 0.89 for manual pivot shift and 0.82 for the mechanized pivot shift test. This study introduced a modified CPM for mechanized pivot shift testing. Although recorded translations and rotations with the mechanized pivot shift test were lower than with manual testing, the clinical advantage of mechanized pivot shift testing is a more repeatable measurement of ATT when compared to manual pivot shift testing. This setup may increase consistency of clinical grading of the pivot shift test. PMID- 20012938 TI - Percutaneous cement augmentation for the treatment of depression fractures of the tibial plateau. AB - The management of insufficiency fractures of the tibial plateau in osteoporotic patients can be very challenging, since it is difficult to achieve a stable fixation, an essential condition for the patients' early mobilization. We present a minimally invasive technique for the treatment of proximal tibial plateau fractures, "tibiaplasty", using percutaneous polymethylmethacrylate augmentation. Five osteoporotic patients (7 fractures) with a non-traumatic insufficiency tibial plateau fracture were treated with this technique at the authors' institution from 2006 to 2008. The patients' median age was 79 (range 62-88) years. The intervention was performed percutaneously under general or spinal anesthesia; after the intervention, immediate full weight bearing was allowed. The technique was feasible in all patients and no complications related to the intervention were observed. All patients reported a relevant reduction in pain, were able to mobilize with full weight bearing and would undergo the operation again. No secondary loss of reduction or progression of arthrosis was observed in radiological controls; no revision surgery was required. Our initial results indicate that tibiaplasty is a good treatment option for the management of insufficiency in tibial plateau fractures in osteoporotic patients. The technique is minimally invasive, safe and allows immediate mobilization without restrictions. In our group of patients, we found excellent early to mid-term results. PMID- 20012939 TI - Increased risk of serious infections in women with osteopenia or osteoporosis treated with denosumab. PMID- 20012941 TI - Arsenic: metabolism and transport mechanisms in human hepatocytes. PMID- 20012940 TI - Gonadal sex steroid status and bone health in middle-aged and elderly European men. AB - SUMMARY: The influence of sex steroids on calcaneal quantitative ultrasound (QUS) parameters was assessed in a population sample of middle-aged and elderly European men. Higher free and total E(2) though not testosterone, were independently associated with higher QUS parameters. INTRODUCTION: The aim of this study was to investigate the association between QUS parameters and sex steroids in middle-aged and elderly European men. METHODS: Three thousand one hundred forty-one men aged between 40 and 79 years were recruited from eight European centres for participation in a study of male ageing: the European Male Ageing Study. Subjects were invited by letter to attend for an interviewer administered questionnaire, blood sample and QUS of the calcaneus (Hologic SAHARA). Blood was assessed for sex steroids including oestradiol (E(2)), testosterone (T), free and bio-available E(2) and T and sex hormone binding globulin (SHBG). RESULTS: Serum total T was not associated with any of the QUS parameters. Free T and both free and total E(2) were positively related to all QUS readings, while SHBG concentrations were negatively associated. These relationships were observed in both older and younger (<60 years) men. In a multivariate model, after adjustment for age, centre, height, weight, physical activity levels and smoking, free E(2) and SHBG, though not free T, remained independently associated with the QUS parameters. After further adjustment for IGF-1, however, the association with SHBG became non-significant. CONCLUSION: Higher free and total E(2) are associated with bone health not only among the elderly but also middle-aged European men. PMID- 20012943 TI - Evaluation of migration behaviour of therapeutic peptide hormones in capillary electrophoresis using polybrene-coated capillaries. AB - Modelling electrophoretic mobility as a function of pH can be simultaneously used for determination of ionization constants and for rapid selection of the optimum pH for separation of mixtures of the modelled compounds. In this work, equations describing the effect of pH on electrophoretic behaviour were used to investigate migration of a series of polyprotic amphoteric peptide hormones between pH 2 and 12 in polybrene-coated capillaries. Polybrene (hexadimethrin bromide) is a polymer composed of quaternary amines that is strongly adsorbed by the fused silica inner surface, preventing undesired interactions between the peptides and the inner capillary wall. In polybrene-coated capillaries the separation voltage must be reversed, because of the anodic electroosmotic flow promoted by the polycationic polymer attached to the inner capillary wall. The possibility of using polybrene-coated capillaries for determination of accurate ionization constants has been evaluated and the optimum pH for separation of a mixture of the peptide hormones studied has been selected. Advantages and disadvantages of using bare fused-silica and polybrene-coated capillaries for these purposes are discussed. PMID- 20012942 TI - Concentration of the macrolide antibiotic tulathromycin in broncho-alveolar cells is influenced by comedication of rifampicin in foals. AB - Macrolide antibiotics penetrate in the lung against steep concentration gradients into the epithelial lining fluid (ELF) and broncho-alveolar cells (BAC). Since they interact with ABCB1, ABCC2, and organic anion transporting proteins (OATPs), which are localized to lung tissue, pulmonary concentration may be influenced by rifampicin (RIF), an inducer and modulator of efflux and uptake transporters. We measured concentrations of tulathromycin (TM) in plasma, ELF and BAC in 21 warm blooded foals 24 and 192 h after first and last intramuscular injection of 2.5 mg/kg TM once weekly for 6 weeks. In 11 foals, TM was combined with RIF (10 mg/kg twice daily), and mRNA expression of ABCB1 and ABCC2 in BAC was assessed before and after RIF. Affinity of TM to ABCB1 and ABCC2 was measured by transport assays using cell monolayers and membrane vesicles of MDCKII and 2008 cells transfected with ABCB1 and ABCC2, respectively. At steady state, TM concentrated manifold in ELF and BAC. Comedication of RIF significantly decreased the AUC of TM (18.5 +/- 4.0 versus 24.4 +/- 3.7 microg x h/ml, p < 0.05) and lowered its concentrations in plasma (24 h, 0.17 +/- 0.05 versus 0.24 +/- 0.05 microg/ml; 192 h, 0.05 +/- 0.01 versus 0.06 +/- 0.01 microg/ml) and BAC (24 h, 0.84 +/- 0.36 versus 1.56 +/- 1.02 microg/ml; 192 h, 0.60 +/- 0.23 versus 1.23 +/- 0.90 microg/ml, all p < 0.05). Treatment with rifampicin did not markedly induce ABCB1 and ABCC2 expression. TM had no affinity to ABCB1 and ABCC2 in vitro. Concentration of TM in the lung of foals was significantly lowered by comedication of rifampicin most likely caused by extrapulmonary mechanisms leading to lower plasma concentrations. PMID- 20012944 TI - Effect of food processing on plant DNA degradation and PCR-based GMO analysis: a review. AB - The applicability of a DNA-based method for GMO detection and quantification depends on the quality and quantity of the DNA. Important food-processing conditions, for example temperature and pH, may lead to degradation of the DNA, rendering PCR analysis impossible or GMO quantification unreliable. This review discusses the effect of several food processes on DNA degradation and subsequent GMO detection and quantification. The data show that, although many of these processes do indeed lead to the fragmentation of DNA, amplification of the DNA may still be possible. Length and composition of the amplicon may, however, affect the result, as also may the method of extraction used. Also, many techniques are used to describe the behaviour of DNA in food processing, which occasionally makes it difficult to compare research results. Further research should be aimed at defining ingredients in terms of their DNA quality and PCR amplification ability, and elaboration of matrix-specific certified reference materials. PMID- 20012945 TI - E-learning in applied instrumental analysis: the example of cultural heritage preservation. PMID- 20012946 TI - Metabolomic investigation of gastric cancer tissue using gas chromatography/mass spectrometry. AB - Gastric cancer screening or diagnosis is mainly based on endoscopy and biopsy. The aim of this study was to identify the difference of metabolomic profile between normal and malignant gastric tissue, and to further explore tumor biomarkers. Chemical derivatization together with gas chromatography/mass spectrometry (GC/MS) was utilized to obtain the metabolomic information of the malignant and non-malignant tissues of gastric mucosae in 18 gastric cancer patients. Acquired metabolomic data was analyzed using the Wilcoxon rank sum test to find the tissue metabolic biomarkers for gastric cancer. A diagnostic model for gastric cancer was constructed using principal component analysis (PCA), and was assessed with receiver-operating characteristic (ROC) curves. Results showed that 18 metabolites were detected differently between the malignant tissues and the adjacent non-malignant tissues of gastric mucosa. Five metabolites were also detected differently between the non-invasive tumors and the invasive tumors. The diagnostic model could discriminate tumors from normal mucosae with an area under the curve (AUC) value of 0.9629, and another diagnostic model constructed for clinical staging was assessed with an AUC value of 0.969. We conclude that the metabolomic profile of malignant gastric tissue was different from normal, and that the selected tissue metabolites could probably be applied for clinical diagnosis or staging for gastric cancer. PMID- 20012949 TI - Bare spot of the glenoid fossa in children: incidence and MRI features. AB - BACKGROUND: The bare spot of the glenoid fossa is a normal cartilage defect seen frequently in adults. It has been used on arthroscopy as a landmark for the center of the glenoid fossa. There are no reports of this variant in children, but we have noted it on some pediatric clinical shoulder MRI studies. OBJECTIVE: Our main purpose is to evaluate the incidence of the bare spot in children and define location and MRI features. MATERIALS AND METHODS: Shoulder MRI studies (total 570) from 2004 to 2008 were reviewed. Children were divided into two age groups: group 1, 0-10 years (n = 200), group 2, 11-20 years (n = 370). RESULTS: A total of 12 bare spots (2.1%) were identified; all were seen in group 2. Eight (67%) were central and four were eccentric in the glenoid fossa. All showed a well-marginated focal cartilage defect containing hyperintense joint fluid or contrast agent. Three also had air. CONCLUSION: The bare spot is seen in children. The absence in children younger than 10 years and the low incidence in the second decade support the proposed acquired nature. Familiarity with this finding is important so as not to misinterpret it as a pathologic condition. PMID- 20012947 TI - Laryngeal somatosensory deficits in Parkinson's disease: implications for speech respiratory and phonatory control. AB - Parkinson's disease (PD) is often associated with substantial impairment of speech respiratory and phonatory control. However, the degree to which these impairments are related to abnormal laryngeal sensory function is unknown. This study examined whether individuals with PD exhibited abnormal and more asymmetric laryngeal somatosensory function compared with healthy controls, and whether these deficits were associated with disease and voice severity. Nineteen PD participants were tested and compared with 18 healthy controls. Testing included endoscopic assessment of laryngeal somatosensory function, with aerodynamic and acoustic assessment of respiratory and phonatory control, and clinical ratings of voice and disease severity. PD participants exhibited significantly abnormal and asymmetric laryngeal somatosensory function compared with healthy controls. Sensory deficits were significantly associated with timing of phonatory onset, voice intensity, respiratory driving pressure, laryngeal resistance, lung volume expended per syllable, disease severity, and voice severity. These results suggest that respiratory and phonatory control are influenced by laryngeal somatosensory function, that speech-related deficits in PD are related to abnormal laryngeal somatosensory function, and that this function may degrade as a function of disease severity. Thus, PD may represent a model of airway sensorimotor disintegration, highlighting the important role of the basal ganglia and related neural networks in the integration of laryngeal sensory input for speech-related motor control. PMID- 20012950 TI - Splenic artery aneurysm: a diagnostic challenge in the setting of extensive portal venous collaterals. AB - We present a 16-year-old boy with autoimmune liver disease and longstanding portal hypertension in whom a CT arteriogram demonstrated a large aneurysm arising from the distal, extra-parenchymal portion of the splenic artery. Because of its location adjacent to multiple venous collaterals, the aneurysm was indistinguishable from splenic varices on initial imaging with Doppler sonography and on portal venous-phase CT. There is an increased risk of rupture of splenic artery aneurysms in the post-liver transplant period, with high associated mortality, and therefore diagnosis of splenic artery aneurysm prior to liver transplantation is clinically important. It is quite possible that the diagnosis of splenic artery aneurysm in this case would have been missed in the absence of dedicated arterial-phase imaging. As radiologists strive to reduce radiation exposure in children, this case highlights a potential diagnostic pitfall of both Doppler sonography and venous or single-acquisition arterial/venous-phase CT angiogram in children with venous collaterals and an undiagnosed splenic artery aneurysm. PMID- 20012952 TI - Hereditary hemorrhagic telangiectasia and juvenile polyposis: an overlap of syndromes. AB - Hereditary hemorrhagic telangiectasia (HHT) (Osler-Weber-Rendu syndrome) is a syndrome characterized by multiorgan telangiectases and arteriovenous malformations. A subset of patients with a mutation in the MADH4 gene on chromosome 18 exhibits an overlapping syndrome of HHT and juvenile polyposis (JPS). We present one such family. Genetic testing is warranted when either HHT or JPS is diagnosed, as early recognition of this syndrome overlap allows appropriate management of these patients. PMID- 20012951 TI - Transient reticular gallbladder wall thickening in severe dengue fever: a reliable sign of plasma leakage. AB - BACKGROUND: Dengue fever (DF) is an acute infection caused by a flavivirus. Although most patients present mild symptoms, some progress to a severe condition characterized by hypovolemic shock and hemorrhagic phenomena. The main feature of this severe form of DF is plasma leakage. Gallbladder wall thickening (GBWT), ascites and pleural effusion represent the sonographic triad of plasma leakage in DF. OBJECTIVE: To evaluate the plasma leakage triad in severe DF with emphasis on the GBWT. MATERIALS AND METHODS: Thirty-seven children with severe DF underwent abdominal US on the day of admittance and on the day of discharge, or 7 days after the first examination if the child was still hospitalized. RESULTS: Of the 37 children, 33 (89.2%) presented GBWT, 29 (78.4%) ascites and 26 (70.3%) pleural effusion. All of these findings had resolved by the second examination. Of the 33 GBWTs, 29 (87.9%) presented a reticular pattern, which could be considered typical of plasma leakage in patients with severe DF. CONCLUSION: GBWT, ascites and pleural effusion are transient findings in DF. The authors have described a typical reticular pattern of GBWT that can be used to diagnose and follow up on patients with severe DF and should not be considered an acalculous cholecystitis. PMID- 20012953 TI - Computed Tomography measurements of the normal and the pathologic cochlea in children. AB - BACKGROUND: Radiological investigation is frequently undertaken to assess the aetiology of sensorineural hearing loss (SNHL). OBJECTIVE: To establish the CT measurements of the normal cochlea in children and to determine radiological criteria correlated with SNHL. MATERIALS AND METHODS: A retrospective study of temporal bone CT performed in 159 children, age range from 3 days to 16 years between February 1999 and July 2004. A control group (n = 88) comprised children without SNHL; the SNHL group comprised 71 children. The width of the second turn of the cochlea (CW), the cochlear height (CH), and the width of the bony canal for the cochlear nerve (WCN) were measured on a reference plane containing the modiolus, the posterior semicircular canal, the footplate, and the stapes arch. RESULTS: Width of the canal measurements or=2.5 mm supported the diagnosis of SNHL with a specificity of 97% and 91%, respectively. Cochlear width was found to be significantly smaller in the SNHL group (5.61 +/- 0.51 mm) than in the control group (5.75 +/- 0.31 mm, P < 0.02), a size <5.4 mm being highly suggestive of SNHL with a specificity of 90%. No significant variations of all measurements were found with age. CONCLUSION: Appropriate measurements of WCN and CW are highly correlated with SNHL. PMID- 20012954 TI - Investigation of stratum corneum lipid model membranes with free fatty acid composition by neutron diffraction. AB - The outermost epidermal layer, the stratum corneum (SC), is the main skin barrier. Studies of SC model systems enable characterization of the influence of individual lipids on the organization of the SC lipid matrix, which is the main pathway of water through the skin. This work presents a neutron diffraction study of the SC model membranes based on short-chain ceramide 6 with nearly realistic composition of free fatty acids (FFA) at physiological temperature of the SC. The influence of FFA and the effect of cholesterol-cholesterol sulfate substitution on the structure and hydration of the SC model membranes are described. The structure of the SC membrane with FFA is close to the structure of the earlier studied SC membrane based on short-chain palmitic acid (PA) and does not vary significantly under changes of the ratio of the main membrane components. FFA accelerates membrane swelling at the same low level of hydration of both PA- and FFA-containing membranes. The substitution of cholesterol sulfate by cholesterol in the membrane composition decreases membrane swelling and leads to phase separation in the model system. PMID- 20012955 TI - Structural correlates of mouse IgA allotypes. AB - A set of mouse IgAs containing amino acids differing amongst the six alpha-chain allotypes was constructed by mutating an S107-IgA plasmid and transfecting it into a non-producer myeloma cell line along with a kappa-chain plasmid. The secreted IgAs were examined for their possession of a covalent bond between alpha and light (L)-chains and for their ability to bind to three anti-allotypic monoclonal antibodies, HIS-M2, HY-15, and HY-16. IgA of the Igh-2(a) allotype was found to be unique in its total lack of a covalent bond between alpha and L: chains, formation of which apparently depends on the presence of an "extra" Cys in the hinges of all of the other five allotypes. The allotypic epitopes are associated with identifiable amino acids in the Calpha1 region of the molecule. Binding to HIS-M2 requires Ala 216 whereas binding to HY-15 requires Pro 216 and Asp 222. Binding to Hy-16 requires Arg 183 and either Pro 216 or Ser 216 but not Ala 216. However, binding to HY-16 by all of the IgAs produced by transfectants is impaired by defective glycosylation in the transfected myeloma and is only revealed after deglycosylation. PMID- 20012956 TI - Paclitaxel combined with ifosfamide in anthracycline- and docetaxel-pretreated metastatic breast cancer: activity independence of prior docetaxel resistance. AB - BACKGROUND: We evaluated the efficacy and tolerability of combined paclitaxel and ifosfamide in anthracycline- and docetaxel-pretreated metastatic breast cancer (MBC). METHODS: Patients received paclitaxel (175 mg/m(2) i.v. in a 3-h infusion) on day 1 and ifosfamide (1.5 g/m(2) i.v. in a 15-min infusion) on days 1-3, every 3 weeks for a maximum of nine cycles. The tumor response was assessed every two cycles. RESULTS: We enrolled 34 patients with a median age of 50 years. Thirty patients had visceral metastases. Anthracycline- and docetaxel-based chemotherapy had previously been administered to 18/13 and 13/21 patients, respectively, in (neo)adjuvant/metastatic settings. Three patients had not previously received anthracycline due to abnormal cardiac functions. A total of 174 cycles of chemotherapy were delivered with a median of six cycles. The response rate under the intent-to-treat analysis was 23.5% (all partial responses) with a median response duration of 14 months. The disease control rate was 70.6%. The median progression-free and overall survival were 5.9 and 8.5 months, respectively. There was no apparent relationship between activity and prior docetaxel resistance. The incidence of grade III/IV neutropenia was 46.6% (81 of 174 cycles) with febrile neutropenia of only 1.7%. Major grade III/IV non hematological toxicities included peripheral neuropathy (6 of 34 patients) and infection (4 of 34 patients). There were no treatment-related deaths. CONCLUSION: Paclitaxel combined with ifosfamide was effective and tolerable in anthracycline /docetaxel-pretreated MBC. Overcoming docetaxel resistance by using paclitaxel in combination with ifosfamide needs to be addressed through further investigation. PMID- 20012957 TI - SCO3900, co-transcripted with three downstream genes, is involved in the differentiation of Streptomyces coelicolor. AB - SCO3900, a putative target gene of the developmental regulator WhiI of Streptomyces coelicolor, encodes a protein similar to PadR from Pediococcus pentosaceus. The SCO3900 disruption mutant exhibited a bald phenotype with sparse aerial hyphae. However, a single copy of SCO3900 under the control of its native promoter could not complement the mutant phenotype. A fragment containing SCO3900 and its downstream three genes completely restored the mutant phenotype to the wild-type. These suggested that the disruption of SCO3900 exerted a polar effect on its downstream genes. The co-transcription of SCO3899, SCO3898, and SCO3897 with SCO3900 was confirmed by the RT-PCR. Moreover, overexpression of SCO3900 in the wild-type strain caused the same phenotype as that of the disruption mutant. The results suggested that SCO3900 encodes a negative regulator participating in morphological differentiation of S. coelicolor by influencing the expression of its downstream genes. PMID- 20012958 TI - Rare localizations of bone sarcoidosis: two case reports and review of the literature. AB - Sarcoidosis is a multisystem disease of unknown origin. Granulomatous bone involvement has an overall incidence of 1-13%. This incidence is probably underestimated in certain patient series because bone involvement is often asymptomatic. The small bones of hands and feet are the most common localizations, while skull, knee, rib, pelvic and sternal localizations are rarely reported. Here we describe some interesting cases of chronic sarcoidosis with unusual bone localizations observed at our regional referral centre for sarcoidosis. We also review the literature to underline the complexity of the disease, the problem of differential diagnosis with respect to malignancies and the need for appropriate and effective therapy of this rare localization. PMID- 20012959 TI - Increased level of tumor necrosis factor-alpha in patients with antiphospholipid syndrome: marker not only of inflammation but also of the prothrombotic state. AB - Connections between inflammation and thrombosis are intriguing, especially in a condition such as an antiphospholipid syndrome (APS), a disease characterized by immune-mediated thrombosis. Tumor necrosis factor alpha (TNF-alpha) is a cytokine which shares proinflammatory and prothrombotic actions, while a soluble form of interlukin-2 receptor (sIL-2R) is considered a typical marker of (auto)immune inflammation with not known direct links to thrombosis. The differences in the pathogenesis of APS as compared to other autoimmune diseases might be connected with different serum levels of both mediators. To answer this question, we studied 147 patients with systemic lupus erythematosus (SLE), 21 with SLE-like syndrome (SLE-LS), 20 with isolated APS (primary antiphospholipid syndrome, PAPS), and 32 healthy controls. Thirty-six patients from the SLE group fulfilled the updated APS criteria (secondary APS, SAPS). In comparison to healthy subjects, TNF-alpha concentration was increased in all patients, while sIL-2R rose significantly in the SLE group only. APS (both SAPS and PAPS) was characterized by the highest levels of TNF-alpha. Moreover, patients with lupus anticoagulant or elevated levels of IgG anticardiolipin or IgG anti-beta(2) glycoprotein I antibodies had higher TNF-alpha levels than patients without the presence of any type of antiphospholipid antibodies (aPL). In conclusion, the presence of aPL is associated with higher TNF-alpha level, whereas increased level of sIL-2R is rather connected with definite SLE where inflammatory processes prevail. It might be hypothesized that TNF-alpha plays a major role in pathogenesis of APS thrombotic phenomena. PMID- 20012960 TI - Report of anti-CCP antibody positive paraneoplastic polyarthritis and review of the literature. AB - A 45-year-old female presented to the rheumatology clinic with complaint of pain and swelling of multiple small joints of the hands and feet. She also complained of cough and shortness of breath onset around the same time. Since her cyclic citrullinated peptide antibody (anti-CCP) and rheumatoid factor tests were positive, rheumatoid arthritis (RA) was diagnosed and she was started on prednisone with plans for additional disease modifying therapy. Chest X-ray showed a small right pleural effusion. While additional pulmonary evaluation was being planned, a few weeks later she presented with dyspnea, fever and tachycardia. Spiral CT showed pulmonary emboli and increased pleural effusion and patient was started on anticoagulation. A chest tube was placed and exudative pleural effusion was drained. Cytology sample from bronchoscopy raised concerns for adenocarcinoma. Open lung biopsy confirmed moderately differentiated adenocarcinoma. The patient died of lung cancer in the hospital 8 weeks from her diagnosis of RA. We describe a case of paraneoplastic polyarthritis with positive anti-CCP antibody test which has not been reported before. We also review the literature on paraneoplastic arthritis which has been described in association with various other malignancies besides lung cancer. PMID- 20012961 TI - Should we routinely evaluate serum alkaline phosphatase levels in patients with gout? AB - Hyperuricemia and gout can be seen in patients with Paget's disease of bone (PD). This may be secondary to increased nucleic acid turnover in accelerated bone remodeling. It is unclear whether all PD patients should be evaluated for the presence of PD and gout in the same patient. In this report, I present a gout patient who later diagnosed as having PD and discuss the relationship between gout and PD. PMID- 20012962 TI - The inadequate anti-tumour necrosis factor (anti-TNF) therapy in patients with ankylosing spondylitis. PMID- 20012963 TI - Unusual central nervous system involvement of rheumatoid arthritis: successful treatment with steroid and azathioprine. AB - Central nervous system involvement of rheumatoid arthritis (RA) frequently develops in patients who had a long-term history of RA, irrespective of the disease activity of systemic arthritis, and it has a high mortality rate despite treatment. Since clinical symptoms and radiologic signs are rather nonspecific, in short of doing biopsy, the diagnosis of rheumatoid meningitis is one of exclusion. However, the strongly positive rheumatoid factor in the cerebrospinal fluid is quite specific. We here report a 70-year-old man who had not been diagnosed as RA before he was admitted with neurological findings, who was diagnosed as RA later and successfully treated with prednisolone and azathioprine. PMID- 20012964 TI - The diagnostic utilities of anti-agalactosyl IgG antibodies, anti-cyclic citrullinated peptide antibodies, and rheumatoid factors in rheumatoid arthritis. AB - The purpose of this study was to investigate the diagnostic utilities of anti agalactosyl IgG antibody (CARF), anti-cyclic citrullinated peptide (CCP) antibody and rheumatoid factor (RF) in rheumatoid arthritis (RA), non-RA rheumatic diseases, and chronic viral hepatitis. The authors determined serum levels of CARF and anti-CCP2 by ELISA and IgM-RF by a immunonephelometric method in 834 controls and in 397 patients with the following conditions: RA (100), non-RA rheumatic diseases [systemic lupus erythematosus (SLE) 30, primary Sjogren's syndrome 18, systemic sclerosis 30, inflammatory myositis 19], chronic viral hepatitis B and C (HBV 100, HCV 100). The sensitivities of CARF (83%) and anti CCP (85%) were significantly higher than that of RF (75%, p = 0.01, respectively) in RA, and the specificity of anti-CCP (98%) was significantly higher than those of CARF (92%) and RF (90%, p < 0.001, respectively). A comparison of receiver operating characteristic (ROC) curves revealed that the diagnostic accuracies of CARF and anti-CCP were superior to that of RF (CARF vs. RF, p = 0.008, anti-CCP vs. RF, p = 0.017) in RA. CARF positivity was significantly higher than those of anti-CCP (p = 0.007) and RF (p = 0.008) in systemic sclerosis, and the positivity of CARF was significantly higher than that of anti-CCP in Sjogren's syndrome (p = 0.016). Furthermore, CARF had significantly higher positivity than anti-CCP or RF in chronic viral hepatitis B and C. Finally, the titers of these three markers in RA were significantly higher than in non-RA rheumatic diseases and in chronic viral hepatitis B and C. Our results suggest that anti-CCP is the most useful serologic marker for the differentiation of RA and non-RA rheumatic diseases, and chronic viral hepatitis B and C. PMID- 20012965 TI - Rapid in planta evaluation of root expressed transgenes in chimeric soybean plants. AB - Production of stable transgenic plants is one factor that limits rapid evaluation of tissue specific transgene expression. To hasten the assessment of transgenes in planta, we evaluated the use of chimeric soybean seedlings expressing transgenic products in roots. Tap roots from four-day old seedlings (cultivars 'Jack' and KS4704) were excised and hairy roots were induced from hypocotyls via Agrobacterium rhizogenes-mediated transformation. Inoculated hypocotyls were screened on a MS-based medium containing either 200 mg/L kanamycin or 20 mg/L hygromycin. Beta-glucuronidase (GUS) activity assay indicated that highest GUS expression was observed in hypocotyls exposed to a 4-d pre-inoculation time, a neutral pH (7.0) for the co-cultivation medium. A 170-bp of the Fib-1 gene and 292-bp of the Y25C1A.5 gene fragments, both related to nematode reproduction and fitness, were cloned independently into pANDA35HK vector using a Gateway cloning strategy. The resulting RNAi constructs of the genes fragments were transformed into soybean using the chimeric hairy root system and evaluated for its effect on soybean cyst nematode (Heterodera glycines) fecundity. Confirmation of transformation was attained by polymerase chain reaction and Southern-blot analysis, and some potential for suppression of H. glycines reproduction was detected for the two constructs. This method takes on average four weeks to produce chimeric plants ready for transgene analysis. PMID- 20012966 TI - Computational approaches for microRNA studies: a review. AB - MicroRNAs (miRNAs) are one class of tiny, endogenous RNAs that can regulate messenger RNA (mRNA) expression by targeting homologous sequences in mRNAs. Their aberrant expressions have been observed in many cancers and several miRNAs have been convincingly shown to play important roles in carcinogenesis. Since the discovery of this small regulator, computational methods have been indispensable tools in miRNA gene finding and functional studies. In this review we first briefly outline the biological findings of miRNA genes, such as genomic feature, biogenesis, gene structure, and functional mechanism. We then discuss in detail the three main aspects of miRNA computational studies: miRNA gene finding, miRNA target prediction, and regulation of miRNA genes. Finally, we provide perspectives on some emerging issues, including combinatorial regulation by miRNAs and functional binding sites beyond the 3'-untranslated region (3'UTR) of target mRNAs. Available online resources for miRNA computational studies are also provided. PMID- 20012967 TI - Identification of novel chromosomal regions associated with airway hyperresponsiveness in recombinant congenic strains of mice. AB - Airway responsiveness is the ability of the airways to respond to bronchoconstricting stimuli by reducing their diameter. Airway hyperresponsiveness has been associated with asthma susceptibility in both humans and murine models, and it has been shown to be a complex and heritable trait. In particular, the A/J mouse strain is known to have hyperresponsive airways, while the C57BL/6 strain is known to be relatively refractory to bronchoconstricting stimuli. We analyzed recombinant congenic strains (RCS) of mice generated from these hyper- and hyporesponsive parental strains to identify genetic loci underlying the trait of airway responsiveness in response to methacholine as assessed by whole-body plethysmography. Our screen identified 16 chromosomal regions significantly associated with airway hyperresponsiveness (genome-wide P 73 cM), chromosome 7 (>63 cM), chromosome 8 (52-67 cM), chromosome 10 (3-7 cM and >68 cM), and chromosome 12 (25-38 cM and >52 cM). Our data identify several likely candidate genes from the 16 regions, including Ddr2, Hc, Fbn1, Flt3, Utrn, Enpp2, and Tsc. PMID- 20012968 TI - Subcutaneous emphysema of the neck after colectomy. PMID- 20012969 TI - Colonic metastasis from a papillary thyroid carcinoma arising in struma ovarii. PMID- 20012970 TI - Double-stapled transanal rectotomy (STARR procedure) for obstructed defecation syndrome seems to have a low degree of acceptance amongst older surgeons. PMID- 20012971 TI - The predictive value of genes of the TGF-beta1 pathway in multimodally treated squamous cell carcinoma of the esophagus. AB - BACKGROUND AND AIM: Pretherapeutic identification of esophageal squamous cell carcinomas (ESCCs) that are likely to respond to neoadjuvant chemoradiotherapy is important in the attempt to improve the prognosis for patients. In the present study, expression of members of the transforming growth factor-beta1 (TGF-beta1) signaling pathway was investigated in pretherapeutic biopsies from 97 ESCCs (cT3, cN0/+, cM0) in patients who underwent neoadjuvant chemoradiotherapy (45 Gy plus cisplatin and 5-fluorouracil) and subsequent esophagectomy in the setting of a single-center prospective treatment trial. MATERIALS AND METHODS: Expression of TGF-beta1 and its downstream effectors Smad4 and Smad7 was assessed using quantitative reverse transcription polymerase chain reaction from RNA prepared from pretherapeutic tumor biopsies. The presence of phosphorylated Smad2 was assessed immunohistochemically. RESULTS: Expression of TGF-beta1 (mean 7.8; range 0.0-25.7 arb. units), Smad4 (mean 0.1; range 0.0-0.4 arb. units), and Smad7 (mean 1.6; range 0.4-16.1 arb. units) varied substantially between the patients. Tumors with total or subtotal regression, as determined by histopathological examination after neoadjuvant chemoradiotherapy, showed significantly higher levels of Smad4 mRNA expression than tumors with minor or no regression (P = 0.032). TGF-beta1 and Smad7 mRNA expression as well as Smad2 protein expression were of no prognostic value. Expression of the four genes under analysis also showed no impact on the overall survival. In contrast, the overall survival correlated significantly with histopathological regression (P < 0.0001) and to a minor degree also with clinical regression grading (P = 0.0254). INTERPRETATION: Among the parameters analyzed, only Smad4 was found to have possible predictive value for esophageal squamous cell carcinoma in patients receiving neoadjuvant chemoradiotherapy. PMID- 20012972 TI - The role of appendicectomy in the subsequent development of inflammatory bowel disease: a UK-based study. AB - BACKGROUND: Several studies have shown an inverse relationship between appendicectomy and subsequent development of inflammatory bowel disease (IBD), although these findings remain contentious. This study aims to further elucidate the role appendicitis/appendicectomy has in the development of IBD. METHOD: All patients undergoing appendicectomy at Hereford County Hospital between 1986 and 2005 were identified from pathology records. Those already diagnosed with IBD were excluded. Age- and sex-matched controls were randomly selected from a database of orthopaedic clinic attendees. Those with prior history of IBD or appendicectomy were excluded. The incidence of IBD was determined by cross referencing with the histology database, colonoscopy database and IBD register. RESULTS: Three thousand eight hundred twenty-nine patients were included in each group, with mean follow-up of 12 +/- 5.9 years. Twelve patients in the appendicectomy group developed IBD. Mean age at diagnosis was 30.3 years, and mean interval from appendicectomy of 3.7 years. Age at appendicectomy and appendicectomy for appendicitis conferred no benefit (Mann-Whitney test, p = 0.991). Eleven patients in the control group developed IBD, with no significant differences in any of the measured outcomes. CONCLUSION: This study has shown no relationship between appendicitis/appendicectomy and development of IBD. PMID- 20012973 TI - [Sjogren's syndrome]. PMID- 20012974 TI - [Current trends in the design and development of monoclonal antibodies against inflammatory mediators for the treatment of rheumatoid arthritis]. AB - As mediators of inflammation, cytokines contribute significantly to both the development and the extent of the inflammatory response in rheumatoid arthritis (RA). In addition, they regulate the differentiation of various cells involved in the pathogenesis of this disease. Tumour necrosis factor alpha (TNF alpha), interleukin (IL)-1 and IL-6 constitute prominent examples of such inflammatory cytokines and have been shown to play an important role in RA. As a consequence, the use of recombinant antibodies targeting these cytokines has revolutionized the treatment of RA. However, a considerable number of RA patients do not respond adequately to therapy with such biologics. Based on this notion, this article summarizes current trends in the design and development of monoclonal antibodies against inflammatory mediators. These include the identification of alterative target structures for anti-cytokine therapies, the specific modification of the antigen-binding CDR of therapeutic antibodies to reduce immunogenicity, alterations of the Fc part and the development of modified antibody fragments to improve the pharmacokinetics and to avoid non-specific immune reactions. Beyond that, efforts are undertaken to optimize the cost of these therapies. PMID- 20012975 TI - [Pathogenesis and diagnosis of Sjogren's syndrome]. AB - Sjogren's syndrome is a common autoimmune disorder. Several genetic risk factors such as STAT-4, ILT6 and the haplotype HLA-B8/DR3 have been identified. In addition, there are environmental risk factors, possibly chronic viral infections. In the pathophysiology of Sjogren's syndrome T and B cells infiltrate the salivary and lacrimal glands. As a consequence of the destruction of glandular cells by cytotoxic T cells, production of cytokines and autoantibodies inhibiting glandular function, the production of saliva and tears is decreased. The feeling of dry eyes and mouth is frequently not noticed by the patients. Therefore, Sjogren's syndrome should also be considered when extraglandular manifestations such as vasculitis, polyneuropathy or arthritis occur, even when the patients do not complain of dry eyes and mouth. Establishing the diagnosis of Sjogren's syndrome requires verification of reduced glandular function, for example using Schirmer's test and the Saxon test. The confirmation of Sjogren's syndrome as a cause of sicca syndrome is subsequently performed by the detection of autoantibodies against Ro (SS-A) and La (SS-B) and/or by a salivary gland biopsy. PMID- 20012976 TI - [Epidemiology of primary Sjorgren's syndrome]. AB - According to the classification criteria of the American-European Consensus Group (AECG), the prevalence of primary Sjogren's syndrome (pSS) of about 0.2% in the adult population and a yearly incidence of 4/100.000 in the general population are far lower than previously assumed. Moreover, the repeatedly reported male/female ratio of 1:9 seems to lie more in the range of 1:20. Male pSS patients show fewer immunological, histopathological or sialographic findings and organ involvement. Information on age at disease onset has also changed over the last decade. Recent studies indicate an onset age of approximately 45 years as compared to 56 in earlier studies of the last decade. Patients with an early disease onset are more frequently positive for rheumatoid factor (RF) and/or anti Ro/SS-A. These patients also seem to have a higher risk of developing hypocomplementemia or lymphadenopathy. As compared to earlier cohorts, the introduction of the rather specific AECG criteria will probably result in the participation of fewer men, younger patients in general and of more seriously ill patients in future cohorts. The change in the spectrum of pSS patients obviously reflects the altered classification criteria since the AECG criteria require anti Ro/La positivity and therefore exclude a high number of patients with other immunological markers who also show severe sicca symptoms and organ involvements. About 5%-10% of pSS patients in rheumatological care suffer from severe extraglandular manifestations, which generally occur soon after disease onset. In particular, palpable purpura, hypocomplementemia, cryoglobulinemia and lymphoma are associated with increased mortality. In Germany, approximately one tenth of Sjogren syndrome patients receive specialized rheumatological care. There is still insufficient knowledge about the vast majority of pSS patients who are not treated by rheumatologists. These patients, as well as all those who, according to the AECG criteria, are not classified as having pSS either due to anti-Ro/La negativity or having secondary Sjogren's syndrome, probably add up to at least 0.4% of the adult population which, at present, suffers from considerable immunopathologic sicca symptoms. PMID- 20012977 TI - [Ophthalmological complications in Sjogren's syndrome]. AB - BACKGROUND: Ocular complications are typical sequels of primary and secondary Sjogren's syndrome. Since these can lead to blindness in the case of insufficient therapeutic steps, diagnostic tools and therapeutic options in Sjogren-syndrome associated ocular diseases are outlined. METHODS: Literature review from PubMed and own clinical and experimental results. RESULTS: Numerous ocular complications can occur both in primary and secondary Sjogren's syndrome. The main problems involve tear film disturbances leading to severe forms of dry eye. CONCLUSION: Early and close cooperation between rheumatologist and ophthalmologist can avoid long-term complications in patients with Sjogren's disease and ocular complications. PMID- 20012978 TI - The Fixion expandable stem hemiarthroplasty for displaced femoral neck fracture: technical features and pilot study. AB - BACKGROUND: Management of displaced femoral neck fracture in elderly patients is challenging due to the patient's high risk profile, poor quality bone stock and muscle weakness. MATERIALS AND METHODS: Fifty-one patients with displaced (Garden 3-4) intracapsular femoral neck fractures were treated with a newly designed Fixion expandable stem hemiarthroplasty implant and followed thereafter for a minimum of 6 months. RESULTS: Thirty patients (58.8%) had concomitant diseases graded 3-4 by the ASA scoring system. Two patients (4.8%) incurred deep wound infections that necessitated hardware removal. One case (2.6%) of dislocation was treated by closed reduction. Forty-two patients (82.4%) survived >6 months and cooperated with the study protocol. Mild groin/thigh pain was reported by 11 patients (26.2%). Twenty-six (61.9%) had lost 1-2 out of 4 grades of mobility. CONCLUSIONS: Fixion implant may be considered a further step in the evolution of femoral joint hemiarthroplasty. In the short term, it has proven to be as effective as cemented implant and to be user-friendly for the surgeon. It is also anticipated to be feasible whenever revision is required. PMID- 20012979 TI - Successful laser therapy of a papular acantholytic dyskeratosis of the vulva: case report and review of literature. AB - BACKGROUND: The papular acantholytic dyskeratosis summarizes a collection of papular skin lesions which occur in intertriginous areas and in the genital area. They show a very characteristic histology without the connection to a syndrome such as the Hailey-Hailey disease or the Darier disease. METHODS: We present the case of an affected 45-year-old woman and evaluate the laser therapy as therapeutic option. RESULTS: A long lasting reduction of the symptoms was achieved by paying special attention to involving the deep tissue layers while performing the laser therapy. Despite causing a long and painful healing process, this was the only way to achieve a lasting reduction of the symptoms. CONCLUSIONS: Laser therapy in afflicted areas can improve the symptoms of the papular acantholytic dyskeratosis. The deep tissue laser therapy showed more success than the superficial laser therapy. PMID- 20012980 TI - Total laparoscopic hysterectomy in 26 min. PMID- 20012981 TI - Effect and safety of high-dose dienogest (20 mg/day) in the treatment of women with endometriosis. AB - PURPOSE: Hormonal treatment of endometriosis is often continued for long periods and has the potential to affect many essential metabolic processes. The current study aimed to determine the effects and safety of high-dose dienogest as a medical endometriosis therapy. METHODS: The effects and safety of high-dose dienogest, 20-30 mg/day for 24 weeks, were examined in 21 women aged 18-52 years with laparoscopically and histologically proven endometriosis stage I-IV (according to revised American Society of Reproductive Medicine criteria). At baseline and week 24, sera were obtained and stored at -20 degrees C prior to analysis. RESULTS: The study showed no clinically significant effect of high-dose dienogest on thyroid or adrenal function, electrolyte balance or haematopoiesis. High-dose dienogest therapy also had no appreciable effects on glucose and lipid metabolism, liver enzymes or haemostasis. For instance, although dienogest mediated small increases in the haemostatic variables prothrombin fragment 1 + 2, antithrombin III and protein C, final levels (at week 24) remained within normal reference ranges for these parameters. The exception was the HDL-3 cholesterol concentration at week 24 (0.97 mmol/l), which increased beyond the normal range of 0.28-0.64 mmol/l. CONCLUSIONS: This investigation yielded a unique dataset on the safety of high-dose dienogest in endometriosis stage I-IV. High-dose dienogest (20-30 mg/day) had little influence upon all the parameters measured. It is therefore likely that lower doses of dienogest would have similarly neutral safety effects: an important consideration in the use of dienogest for the treatment of endometriosis. PMID- 20012982 TI - Exhaled nitric oxide levels are elevated in persons with tetraplegia and comparable to that in mild asthmatics. AB - The role of airway inflammation in mediating airflow obstruction in persons with chronic traumatic tetraplegia is unknown. Measurement of the fraction of exhaled nitric oxide (FeNO) affords a validated noninvasive technique for gauging the airway inflammatory response in asthma, although it has never been assessed in persons with tetraplegia. This study was designed to determine the FeNO in individuals with chronic tetraplegia compared with that in patients with mild asthma and healthy able-bodied individuals. Nine subjects with chronic tetraplegia, seven subjects with mild asthma, and seven matched healthy able bodied controls were included in this prospective, observational, pilot study. All subjects were nonsmokers and clinically stable at the time of study. Spirometry was performed on all participants at baseline. FENO was determined online by a commercially available closed circuit, chemiluminescence method, using a single-breath technique. Subjects with tetraplegia had significantly higher values of FeNO than controls (17.72 +/- 3.9 ppb vs. 10.37 +/- 4.9 ppb; P < or = 0.01), as did subjects with asthma (20.23 +/- 4.64 ppb vs. 10.37 +/- 4.9 ppb, P < or = 0.001). There was no significant difference in FeNO between subjects with tetraplegia and those with asthma (17.72 +/- 3.9 ppb vs. 20.23 +/- 4.64 ppb, P < or = 0.27). Individuals with chronic tetraplegia have FeNO levels that are comparable to that seen in mild asthmatics and higher than that in healthy able-bodied controls. The clinical relevance of this observation has yet to be determined. PMID- 20012983 TI - Linear discriminant analysis and artificial neural network for glaucoma diagnosis using scanning laser polarimetry-variable cornea compensation measurements in Taiwan Chinese population. AB - BACKGROUND: To determine whether linear discriminant analysis (LDA) and artificial neural network (ANN) can improve the differentiation between glaucomatous and normal eyes in a Taiwan Chinese population, based on the retinal nerve fiber layer thickness measurement data from scanning laser polarimetry variable corneal compensation (GDx VCC). METHODS: This study comprised 79 glaucoma (visual field defect, mean deviation: -5.60 +/- 4.23 dB) and 86 healthy subjects (visual field defect, mean deviation: -1.44 +/- 1.72 dB). Each patient received complete ophthalmological evaluation, standard automated perimetry (SAP), and GDx VCC exam. One eye per subject was considered for further analysis. The area under the receiver operating characteristics (AROC) curve, sensitivity, specificity and the best cut-off value for each parameter were calculated. The diagnostic performance of artificial neural network (ANN) and linear discriminant analysis (LDA) for glaucoma detection using GDx VCC measurements will be compared in this study. RESULTS: The individual parameter with the best AROC curve for differentiating between normal and glaucomatous eye was nerve fiber indicator (NFI, 0.932). The highest AROCs for the LDA and ANN methods were 0.950 and 0.970 respectively. CONCLUSION: NFI, ANN and LDF method demonstrated equal diagnostic power in glaucoma detection in a Taiwan Chinese population. PMID- 20012984 TI - The repeated bout effect of eccentric exercise is not associated with changes in voluntary activation. AB - The aim of this study was to compare the possible changes in muscle activation level between a first and second bout of damaging eccentric exercise performed at 2 weeks interval (i.e. repeated bout effect). To that purpose, ten physically active males took part in this study. The eccentric exercise consisted of 10 sets of 12 maximal voluntary contractions (MVC) produced by the knee extensors during movements performed at a constant speed of 160 degrees s(-1). Changes in voluntary and electrically evoked torque in concentric and/or isometric conditions were assessed at the following time points: pre-exercise, and 2 min, 1 and 24 h after each eccentric exercise. At the same time points, voluntary activation was quantified by the superimposed electrical stimulation technique. Muscle soreness and plasma CK activity were measured within 48 h after the eccentric exercise. The results showed that the decrease in eccentric peak torque was linear throughout the exercise protocol. At the end of bouts 1 and 2, torque was significantly reduced by 27.7 +/- 9.1 and 23.4 +/- 11.2, respectively, with no difference between bouts (P > 0.05). At 24 h post-exercise, a lower reduction (P < 0.05) in MVC (17.8 +/- 5.4%) and electrically evoked (16.7 +/- 4.6%) isometric torque was observed for bout 2. In contrast, no statistical difference was found in the deficit in voluntary activation between the two bouts. In conclusion, our results indicate that the repeated bout effect of eccentric exercise appears to reduce muscle damage, but does not influence the level of voluntary activation. PMID- 20012986 TI - Genetic and biochemical analysis of the TLA1 gene in Chlamydomonas reinhardtii. AB - The Chlamydomonas reinhardtii genomic DNA database contains a predicted open reading frame (ORF-P) without an apparent stop-codon and unknown coding sequence, located in close proximity and immediately upstream of the TLA1 gene (GenBank Accession No. AF534570). The latter was implicated in the regulation of the light harvesting chlorophyll antenna size of photosynthesis (Tetali et al. Planta 225:813-829, 2007). To provide currently lacking information on ORF-P and its potential participation in TLA1 gene expression, thus in the regulation of the chlorophyll antenna size, genetic and biochemical analyses were undertaken. The coding and UTR regions of the ORF-P were defined and delineated from those of the adjacent TLA1 gene. ORF-P is shown to encode a protein with a distinct RING-like zinc finger domain that is present in numerous eukaryotic proteins, believed to play a role in cellular ubiquitination, leading to regulation of cellular processes like signaling, growth, transcription, and DNA repair. It is further shown that the two genes share a 74-bp overlap between the 3' UTR region of ORF-P and the 5' UTR region of TLA1. However, they possess distinct start and stop codons and separate coding sequences, and transcribed as separate mRNAs without any trans-splicing between them. Complementation experiments showed that the TLA1 gene alone is sufficient to rescue the truncated chlorophyll antenna size phenotype of the tla1 mutant. Protein sequence alignments in C. reinhardtii and the colorless microalga Polytomella parva suggested that TLA1 defines the relationship between nucleus and organelle in microalgae, indirectly affecting the development of the chlorophyll antenna size. PMID- 20012987 TI - Belowground volatiles facilitate interactions between plant roots and soil organisms. AB - Many interactions between organisms are based on the emission and perception of volatiles. The principle of using volatile metabolites as communication signals for chemo-attractant or repellent for species-specific interactions or mediators for cell-to-cell recognition does not stop at an apparently unsuitable or inappropriate environment. These infochemicals do not only diffuse through the atmosphere to process their actions aboveground, but belowground volatile interactions are similarly complex. This review summarizes various eucaryotes (e.g., plant (roots), invertebrates, fungi) and procaryotes (e.g., rhizobacteria) which are involved in these volatile-mediated interactions. The soil volatiles cannot be neglected anymore, but have to be considered in the future as valuable infochemicals to understand the entire integrity of the ecosystems. PMID- 20012988 TI - Morphological and immunophenotypic features of primary and metastatic giant cell tumour of bone. AB - Giant cell tumour of bone (GCTB) is a primary tumour of bone that may rarely, in the absence of malignant cytological features, produce metastatic lesions, most commonly in the lungs. Whether these lung nodules represent true neoplastic secondaries or implants derived from the primary tumour is not certain. In this study, we have analysed the morphological and immunophenotypic features of 19 conventional GCTBs and corresponding lung nodules for expression of macrophage, osteoclast, proliferation and tumour-associated markers. A striking morphological feature of all GCTBs that produced lung secondaries was the presence of large areas of haemorrhage and thrombus formation; mononuclear and multinucleated cells of GCTB were frequently found within these areas of haemorrhage and thrombus. A similar pattern of CD14, CD33, HLA-DR and CD51 expression was seen in macrophages and giant cells in primary and secondary tumours. Smooth muscle actin expression was frequently noted in primary GCTBs that recurred and metastasised. No difference was seen in the expression of p53, p63, Ki-67, cyclin D1 or Bcl-2 in primary and secondary tumours. Our findings suggest that most lung nodules associated with primary conventional GCTBs are implants derived from tumour emboli formed in areas of haemorrhage and thrombus formation within the primary tumour. PMID- 20012989 TI - Tumor cells infected with oncolytic influenza A virus prime natural killer cells for lysis of resistant tumor cells. AB - Tumor resistance to lysis by resting natural killer (NK) cells may be overcome by priming of NK cells with cytokines or by binding of NK activating receptors to ligands expressed on target cells. In this study, major histocompatibility complex class I (MHC-I)-negative LNCaP and MHC-I-positive DU145 cells were infected with genetically modified influenza A virus lacking the non-structural gene 1 (NS1 IAV). The cells were used to investigate the influence of NS1 IAV infection on NK cell lysis of tumor cells as well as to prime NK cells for lysis of LNCaP and DU145 cells. While LNCaP cells infected with DeltaNS1 IAV showed enhanced lysis when compared with mock-infected cells (93% +/- 1.47 vs. 52% +/- 0.74), both mock-infected and DeltaNS1 IAV-infected DU145 cells were resistant to NK cell lysis. Moreover, NK cells primed with DeltaNS1 IAV-infected LNCaP/DU145 cells effectively lysed resistant DU145 and sensitive LNCaP cells to a greater extent than NK cells primed with mock-infected LNCaP/DU145 or non-primed NK cells. Also, NK cell priming with DeltaNS1 IAV-infected tumor cells enhanced extracellular signal-regulated kinase phosphorylation and increased granule release in NK cells. The increased granule release was specifically mediated by NKp46, which eventually potentiated NK cells primed with DeltaNS1 IAV-infected tumor cells to overcome the inhibitory effects posed by MHC-I expression on DU145 cells. These findings show that in addition to direct lytic activity of NK cells, DeltaNS1 IAV may influence anti-tumoral responses by priming NK cells. PMID- 20012990 TI - Artemisinin derivatives for treatment of uncomplicated Plasmodium falciparum malaria in Sudan: too early for too much hope. AB - The artemisinin-based combination therapy (ACT) is adopted by several countries as first line for malaria treatment in the last decade. Concomitantly, the World Health Organization and other research reports showed a dramatic decline in malaria burden in terms of morbidity, mortality and treatment failure (TF). The optimistic features of ACT are regularly reported with great hopes, while the pessimistic facets either not existing or underreported. However, the dependence on ACT as a single chemotherapeutic agent for malaria control bears considerable risks. Occurrence and spread of artemisinin derivatives (AD) TF will be a major threat, whether it is due to parasite drug resistance or use of poor drug quality. In addition, the safety of AD is not yet fully known. In this short review, two clinical trials performed to evaluate the efficacy and safety of AD, dihydroartemisinin (DHA) plus chloroquine and artesunate (AS) plus fansidar, in Sudan are critically discussed. The conclusions from both studies were that, the TF rate of DHA indicates arrival of counterfeit AD to Africa, and both rate of TF and undesirable effects of AS/SP were recognized. Both findings indicate that it is too early for too much hope on AD. PMID- 20012991 TI - Acanthamoeba strains show reduced temperature tolerance after long-term axenic culture. AB - Acanthamoeba is a genus of free-living organisms that can be found in various habitats. We investigated the physiological characteristics of 15 Acanthamoeba isolates, representing five genotypes (T4, T5, T6, T7, and T11) of both clinical and nonclinical origins. Moreover, in order to evaluate possible alterations from long-term culture, old and fresh isolates were included, and results were compared to a previous study. We found that there is no significant difference in physiological characteristics between genotypes. However, Acanthamoeba strains that had been grown in axenic culture over long periods of time adapted to axenic growth. Overall growth rates under-agarose migration and particularly, temperature tolerance decrease after long-term axenic culture at room temperature. The only trait that remained rather constant was the cytopathic effect. PMID- 20012992 TI - The pathway-specific regulator AveR from Streptomyces avermitilis positively regulates avermectin production while it negatively affects oligomycin biosynthesis. AB - The function of the regulatory protein AveR in Streptomyces avermitilis was examined. An aveR deletion mutant abolished avermectin production and produced more oligomycin, and its phenotype was complemented by a single copy of the aveR gene. Removal of the C-terminal HTH domain of AveR abolished avermectin biosynthesis, indicating the importance of HTH domain for AveR function. Promoter titration and promoter probe assays suggested that the transcription of aveA1, encoding polypeptide AVES1 of avermectin PKS, was activated by AveR. Chromatin immunoprecipitation (ChIP) assay showed that the predicted promoter regions of both the ave cluster and the olm cluster were target sites of AveR, and the DNA binding activity of AveR was dependent on its HTH domain. RT-PCR analysis revealed that the transcriptions of ave structural genes were dependent on AveR, but that of olm structural genes and putative pathway-specific regulatory genes increased in the aveR mutants. Consistent with these observations, overexpression of aveR successfully increased avermectin production. These results indicated that aveR encodes a pathway-specific activator essential for avermectin biosynthesis and it also negatively affects oligomycin biosynthesis. PMID- 20012993 TI - [What is the significance of vitamins for the eye]. PMID- 20012995 TI - Diabetic kidney disease: act now or pay later. PMID- 20012996 TI - On the relationship between glomerular filtration rate and serum creatinine in children. AB - The Schwartz formula (eGFR = kL/Scr, with k = 0.55) to determine the estimated glomerular filtration rate (eGFR) in children with chronic kidney disease (CKD), based on length (L) and serum creatinine (Scr) has recently been updated for enzymatic serum creatinine concentrations, resulting in k = 0.413. Based on a meta-analysis, we evaluated the validity of this updated equation and other published equations for healthy children. This is the first time that publicly available data for healthy children of uncorrected and body surface area (BSA) corrected median GFR have been combined with median serum creatinine values and median lengths and weights from different sources in the literature to evaluate several statistical models to estimate GFR in children. For enzymatic serum creatinine, we show that the simple model for uncorrected GFR (uGFR = k'L(3)/Scr, with k' = 1.32 x 10(-5)) and the BSA-corrected GFR (cGFR = kL/Scr, analogous to the Schwartz formula), with an important age-dependent adaptation for k (k = 0.0414 x 1n (Age) + 0.3018), correlate extremely well with chromium-51 ethylenediamine tetra-acetic acid ((51)Cr-EDTA) data for children between 1 month and 14 years of age. With this age-dependent modification for k, presented here, the simple bedside calculation tool derived by Schwartz can be used for screening all children for CKD. When height information is not available, the Lund-Malmo equation is an excellent alternative. PMID- 20012997 TI - FGF-23 in bone biology. AB - Recent studies have demonstrated that levels of fibroblast growth factor 23 (FGF 23), a key regulator of phosphorus and vitamin D metabolism, rise dramatically as renal function declines and may play a key initiating role in disordered mineral and bone metabolism in patients with chronic kidney disease (CKD). The physiologic importance of FGF-23 in mineral metabolism was first identified in human genetic and acquired rachitic diseases and further characterized in animal models. FGF-23 and its regulators, including phosphate regulating endopeptidase homolog, dentin matrix 1 (DMP1), and matrix extracellular phosphoglycoprotein, are made primarily in bone, specifically in osteocytes. Dysregulation of these proteins results in osteomalacia, implicating the osteocyte in the regulation of skeletal mineralization. Studies in pediatric patients with CKD, the majority of whom have altered skeletal mineralization in early stages of CKD, have demonstrated that skeletal expression of both FGF-23 and its regulator, DMP1, are increased in early stages of CKD and that expression of these proteins is associated with alterations in skeletal mineralization. Thus, dysregulation of osteocytic proteins occur very early in the course of CKD and appear to be central to altered bone and mineral metabolism in this patient population. PMID- 20012998 TI - Very preterm birth is a risk factor for increased systolic blood pressure at a young adult age. AB - Children born very prematurely who show intrauterine growth retardation (IUGR) are suggested to be at risk of developing high blood pressure as adults. Renal function may already be impaired by young adult age. To study whether very preterm birth affects blood pressure in young adults, we measured 24-h ambulatory blood pressure (Spacelabs 90207 device) and renin concentration in 50 very premature individuals (<32 weeks of gestation), either small (SGA) or appropriate (AGA) for gestational age (21 SGA, 29 AGA), and 30 full-term controls who all were aged 20 years at time of measurement. The mean (standard deviation) daytime systolic blood pressure in SGA and AGA prematurely born individuals, respectively, was 122.7 (8.7) and 123.1 (8.5) mmHg. These values were, respectively, 3.6 mmHg [95% confidence interval (CI) -0.9 to 8.0] and 4.2 mmHg (95% CI 0.4-8.0) higher than in controls [119.6 (7.6)]. Daytime diastolic blood pressure and nighttime blood pressure did not differ between groups. We conclude that individuals born very preterm have higher daytime systolic blood pressure and higher risk of hypertension at a young adult age. PMID- 20012999 TI - The cancer anorexia/weight loss syndrome: exploring associations with single nucleotide polymorphisms (SNPs) of inflammatory cytokines in patients with non small cell lung cancer. AB - OBJECTIVE: The cancer anorexia/weight loss syndrome commonly occurs in patients with non-small cell lung cancer (NSCLC) and is characterized by loss of weight and appetite as well as diminished survival. The current study explored whether any of 22 single nucleotide polymorphisms (SNPs) of certain previously implicated inflammatory cytokines (interleukin-1 beta, interleukin-1RN, interleukin-6, and tumor necrosis factor) are associated with this syndrome. PATIENTS AND METHODS: All NSCLC patients who had been enrolled in the Mayo Clinic Lung Cancer Cohort, had completed a health-related questionnaire approximately 6 months after enrollment, and had blood drawn were included in this study, thus yielding a sample size of 471 patients. RESULTS: Sixty-six (14%) patients manifested weight loss shortly after diagnosis, and 152 (32%) reported appetite loss. Only tumor necrosis factor alpha rs800629 was associated with anorexia (odds ratio: 0.46; 95% confidence interval: 0.29, 0.72; p < 0.001); patients who were heterozygous and minor homozygous were less likely to suffer anorexia. Otherwise, there were no statistically significant associations between any of the other 21 SNPs and weight loss and/or anorexia. In univariate analyses, weight loss, anorexia, more advanced cancer stage, and interleukin-1 beta rs1143627 were associated with a worse survival, and interleukin-6 rs2069835 was associated with better survival. However, in multivariate analyses, cancer stage and patient age were the only statistically significant predictors of worse survival. CONCLUSION: No specific SNP was associated with all aspects of the cancer anorexia/weight loss syndrome, but rs800629 may merit further study in cancer-associated anorexia. PMID- 20013001 TI - Preliminary study: reliability of the spinal wheel. A novel device to measure spinal postures applied to sitting and standing. AB - Postural re-education is an integral part of physiotherapy management in patients with back pain. Although the link between posture and back pain is largely unknown, postural re-education is performed on the premise of optimizing spinal alignment to minimize stresses on the passive structures of the spine, to facilitate optimal muscular support and thus to prevent possible damage and further pain. A reliable and clinically meaningful measurement of spinal postures to monitor such interventions remains challenging. This study evaluated within day (intra-tester, inter-tester) and between-day (test-retest) reliability of a novel spinal wheel device measuring thoracic and lumbar postures during sitting and standing. 17 healthy volunteers (age 39.5 +/- 5.4, BMI 25 +/- 9.2; 9 males) were measured three times, by three testers, on two separate occasions (1 week apart). The angular change between C7 and T12 and between T12 and S1 provided thoracic and lumbar curvatures, respectively. Intra-class correlation coefficient (ICC) with 95% confidence intervals and typical error were calculated. Excellent reliability was demonstrated with intra-tester ICCs between 0.947 and 0.980 and typical error between 1.7 degrees and 3.7 degrees and inter-tester ICCs between 0.949 and 0.986 and typical error between 2.0 degrees and 4.7 degrees. Test retest reliability was high with ICCs 0.719-0.908 and typical error between 4.0 degrees and 7.4 degrees. In conclusion, the spinal wheel demonstrated excellent within-day and high between-day reliability. The device may be used in conjunction with 2D camcorder to provide clinically useful visual evaluation of postures for assessment, intervention monitoring, and feedback during postural re education. PMID- 20013002 TI - Dural lesions in lumbar disc herniation surgery: incidence, risk factors, and outcome. AB - In lumbar disc herniation surgery, dural lesions seem to be the most common complication today. Studies on incidence of and outcome after a dural lesion are mainly based on retrospective studies. In a prospective study within the framework of the Swedish Spine Register, 4,173 patients operated on for lumbar disc herniation were evaluated using pre- and 1-year postoperative protocols and complication registration. Mean patient age was 41 (18-81) years and 53% of the patients were male. 93% of the operations were performed on the two lowermost lumbar levels. The incidence of dural lesions in the material was 2.7%. In patients with previous disc surgery, the incidence was doubled, 5%, a significant increase (P = 0.02). Patients with dural lesions preoperatively had more back pain and inferior scores in general health and role emotional domains of the SF 36. These factors, however, were because they had been operated on previously, not related to the dural lesion as such. The relative improvement after surgery was similar whether a dural lesion had occurred or not. It is concluded that a dural lesion is a technical complication which must be solved at the time of surgery but which does not bear any negative implications on the long-term outcome for the patient. PMID- 20013003 TI - Letter to the Editor concerning "The single transoral approach for Os odontoideum with irreducible atlantoaxial dislocation" by Wang X, Fan CY, Liu ZH, Eur Spine J. 2009 Jul 14. [Epub ahead of print]. PMID- 20013004 TI - Single-stage closing-opening wedge osteotomy of spine to correct severe post tubercular kyphotic deformities of the spine: a 3-year follow-up of 17 patients. AB - The correction of severe post-tubercular kyphosis (PTK) is complex and has the disadvantage of being multiple staged with a high morbidity. Here, we describe the procedure and results of closing-opening osteotomy for correction of PTK which shortens the posterior column and opens the anterior column appropriately to correct the deformity without altering the length of the spinal cord. Seventeen patients with PTK (10 males; 7 females) with an average age of 18.3 +/- 10.6 years (range 4-40 years) formed the study group. There were ten thoracolumbar, one lumbar and six thoracic deformities. The number of vertebrae involved ranged from 2 to 5 (average 2.8). Preoperative kyphosis averaged 69.2 degrees +/- 25.1 degrees (range 42 degrees -104 degrees ) which included ten patients with deformity greater than 60 degrees . The average vertebral body loss was 2.01 +/- 0.79 (range 1.1-4.1). The neurological status was normal in 13 patients, Frankel's grade D in three patients and grade C in one. Posterior stabilization with pedicle screw instrumentation was followed by a preoperatively calculated wedge resection. Anterior column reconstruction was performed using rib grafts in four, tricortical iliac bone graft in five, cages in six, and bone chips alone and fibular graft in one patient each. Average operating time was 280 min (200-340 min) with an average blood loss of 820 ml (range 500-1,600 ml). The postoperative kyphosis averaged 32.4 degrees +/- 19.5 degrees (range 8 degrees 62 degrees ). The percentage correction of kyphosis achieved was 56.8 +/- 14.6% (range 32-83%). No patient with normal preoperative neurological status showed deterioration in neurology after surgery. The last follow-up was at an average of 43 +/- 4 months (range 32-64 months). The average loss of correction at the last follow-up was 5.4 degrees (range 3 degrees -9 degrees ). At the last follow-up, the mean preoperative pain visual analogue scale score decreased significantly from 9.2 (range 8-10 points) to 1.5 (range 1-2 points). There was also a significant decrease in mean preoperative Oswestry's Disability Index from 56.4 (range 46-68) to 10.6 (range 6-15). Complications were superficial wound infections in two, neurological deterioration in one, temporary jaundice in one and implant failure requiring revision in one. Single-stage closing-opening wedge osteotomy is an effective method to correct severe PTK. The procedure has the advantage of being a posterior only, single-stage correction, which allows for significant correction with minimal complications. PMID- 20013005 TI - Reconstruction of vein of Labbe in temporo-occipital meningioma invading transverse sinus: technical report. AB - OBJECTIVE AND IMPORTANCE: Meningiomas invading major venous sinuses and veins are not uncommon. However, meningiomas involving both the transverse sinus and vein of Labbe are exceedingly rare. Venous reconstruction can be challenging after radical removal of the meningioma which invades major venous sinuses and veins. In this report, we present reconstruction of vein of Labbe in the surgery of temporo-occipital meningioma invading the transverse sinus. CLINICAL PRESENTATION: A 55-year-old woman was admitted to our institution with a 13-year history of focal epilepsy in the form of transient aphasia with vertigo. Cranial magnetic resonance imaging revealed a homogeneously contrast enhancing lesion measuring 40 mm in diameter in the left temporo-occipital area. Cerebral magnetic resonance venography and venous phase of cerebral digital subtraction angiography revealed invasion of the upper wall of left transverse sinus and total occlusion of the lumen of the vein of Labbe. TECHNIQUE: After removal of all the tumors except the piece attached to the transverse sinus and vein of Labbe with standard neurosurgical techniques, the left transverse sinus lumen and vein of Labbe lumen were explored. The tumor tissue inside the transverse sinus and vein of Labbe were removed along with tumor-invaded venous wall. Venous reconstruction of the transverse sinus and vein of Labbe was performed using autologous dura mater as a patch for the defect. CONCLUSION: This is the first report in the literature in which the vein of Labbe was reconstructed in the surgery of temporo-occipital meningioma invading the vein of Labbe and transverse sinus together. PMID- 20013006 TI - Synovial cyst that compressed the peroneal nerve: case report. AB - We report a case of synovial cyst that induced the compression of the peroneal nerve. The patient presented with foot drop and impossibility to stand on his right heel associated with numbness of anterolateral surfaces of the right shin and the dorsal surface of the foot within a 1-month period. Based on the clinical examination and electroneuromyography data, the compression of nerve by soft elastic mass was confirmed. We treated the patient by total microsurgical excision of the soft elastic mass adherent to the right peroneal nerve at the top third of the patient's shin. Histological study revealed the cyst of synovial membrane. During the postoperative period, a force in extensors of the right foot was restored. On the seventh day, the patient began to step on his right heel, while the zone of tenderness and temperature hypoesthesia in the area of the peroneal nerve innervation remained. PMID- 20013007 TI - Direct switch from levodopa/benserazide or levodopa/carbidopa to levodopa/carbidopa/entacapone in Parkinson's disease patients with wearing-off: efficacy, safety and feasibility--an open-label, 6-week study. AB - The study objective was to assess the efficacy, safety and feasibility of switching from levodopa/benserazide (LB) or levodopa/carbidopa (LC) to levodopa/carbidopa/entacapone (LCE) in Parkinson's disease (PD) patients with wearing-off. This was a multicenter, open-label, 6-week study; the primary outcome was success rate based on the patient-assessed Clinical Global Impression of Change (P-CGI-C). Secondary outcomes included investigator-assessed CGI-C (I CGI-C), change from baseline in Unified Parkinson's Disease Rating Scale (UPDRS), motor/non-motor wearing-off symptoms and quality of life-visual analog scale (QoL VAS). After switching to LCE, 77% of patients reported an 'improvement' (p < 0.0001 vs. patients reporting 'no change or worsening'). Significant improvements were seen in I-CGI-C, UPDRS and QoL-VAS, regardless of prior therapy. Oral levodopa dosing was increased in 28% of patients; the primary outcome remained significant when these patients were excluded. The data suggest that switching from LB/LC to LCE provided a significant benefit in PD patients with wearing-off. PMID- 20013008 TI - Somatosensory responses in normal aging, mild cognitive impairment, and Alzheimer's disease. AB - As a part of a larger study of normal aging and Alzheimer's disease (AD), which included patients with mild cognitive impairment (MCI), we investigated the response to median nerve stimulation in primary and secondary somatosensory areas. We hypothesized that the somatosensory response would be relatively spared given the reported late involvement of sensory areas in the progression of AD. We applied brief pulses of electric current to left and right median nerves to test the somatosensory response in normal elderly (NE), MCI, and AD. MEG responses were measured and were analyzed with a semi-automated source localization algorithm to characterize source locations and timecourses. We found an overall difference in the amplitude of the response of the primary somatosensory source (SI) based on diagnosis. Across the first three peaks of the SI response, the MCI patients exhibited a larger amplitude response than the NE and AD groups (P < 0.03). Additional relationships between neuropsychological measures and SI amplitude were also determined. There was no significant difference in amplitude for the contralateral secondary somatosensory source across diagnostic category. These results suggest that somatosensory cortex is affected early in the progression of AD and may have some consequence on behavioral and functional measures. PMID- 20013009 TI - Overexpression of ornithine decarboxylase increases myogenic potential of H9c2 rat myoblasts. AB - Myoblast differentiation into multinuclear myotubes implies the slow-down of their proliferative drive and the expression of myogenin, an early marker of myogenic differentiation. Natural polyamines-such as putrescine, spermidine and spermine-are low molecular weight organic polycations, well known as mediators involved in cell homeostasis. Many evidences in the literature point to their role in driving cellular differentiation processes. Here, we studied how polyamines may affect the differentiation of the myogenic cell line H9c2 into the muscle phenotype. Cell cultures were committed via a 7-day treatment with insulin which induced increase in the activity of ornithine decarboxylase, the first enzyme in the polyamine biosynthetic pathway, consistent with myogenic differentiation. To evaluate the role of polyamines in the differentiation process, cells were transfected with a plasmid overexpressing a stable ornithine decarboxylase, under control of a constitutive promoter. Overexpressing cells spontaneously differentiate into myotubes, without the need for induction with insulin; multinuclear myotubes and myogenin expression were apparent within 2 days of confluency of cultures. Polyamine depletion-by means of alpha difluoromethylornithine, an irreversible inhibitor of ornithine decarboxylase abolished the differentiation process. These observations support the evidence that polyamines are a key step involved in differentiation of muscle cells. PMID- 20013010 TI - Effects of S-adenosylmethionine decarboxylase, polyamines, amino acids, and weak bases (amines and ammonia) on development and ribosomal RNA synthesis in Xenopus embryos. AB - We have been studying control mechanisms of gene expression in early embryogenesis in a South African clawed toad Xenopus laevis, especially during the period of midblastula transition (MBT), or the transition from the phase of active cell division (cleavage stage) to the phase of extensive morphogenesis (post-blastular stages). We first found that ribosomal RNA synthesis is initiated shortly after MBT in Xenopus embryos and those weak bases, such as amines and ammonium ion, selectively inhibit the initiation and subsequent activation of rRNA synthesis. We then found that rapidly labeled heterogeneous mRNA-like RNA is synthesized in embryos at pre-MBT stage. We then performed cloning and expression studies of several genes, such as those for activin receptors, follistatin and aldolases, and then reached the studies of S-adenosylmethionine decarboxylase (SAMDC), a key enzyme in polyamine metabolism. Here, we cloned a Xenopus SAMDC cDNA and performed experiments to overexpress the in vitro-synthesized SAMDC mRNA in Xenopus early embryos, and found that the maternally preset program of apoptosis occurs in cleavage stage embryos, which is executed when embryos reach the stage of MBT. In the present article, we first summarize results on SAMDC and the maternal program of apoptosis, and then describe our studies on small molecular-weight substances like polyamines, amino acids, and amines in Xenopus embryos. Finally, we summarize our studies on weak bases, especially on ammonium ion, as the specific inhibitor of ribosomal RNA synthesis in Xenopus embryonic cells. PMID- 20013012 TI - Polyamines in biological systems. PMID- 20013011 TI - Polyamines: fundamental characters in chemistry and biology. AB - Polyamines are small cationic molecules required for cellular proliferation and are detected at higher concentrations in most tumour tissues, compared to normal tissues. Agmatine (AGM), a biogenic amine, is able to arrest proliferation in cell lines by depleting intracellular polyamine levels. It enters mammalian cells via the polyamine transport system. Agmatine is able to induce oxidative stress in mitochondria at low concentrations (10 or 100 microM), while at higher concentrations (e.g. 1-2 mM) it does not affect mitochondrial respiration and is ineffective in inducing any oxidative stress. As this effect is strictly correlated with the mitochondrial permeability transition induction and the triggering of the pro-apoptotic pathway, AGM may be considered as a regulator of this type of cell death. Furthermore, polyamine transport is positively correlated with the rate of cellular proliferation. By increasing the expression of antizyme, a protein that inhibits polyamine biosynthesis and transport, AGM also exhibits a regulatory effect on cell proliferation. Methylglyoxal bis(guanylhydrazone) (MGBG), a competitive inhibitor of S-adenosyl-L: -methionine decarboxylase, displaying anticancer activity, is a structural analogue of the natural polyamine spermidine. MGBG has been extensively studied, preclinically as well as clinically, and its anticancer activity has been attributed to the inhibition of polyamine biosynthesis and also to its effect on mitochondrial function. Numerous findings have suggested that MGBG might be used as a chemotherapeutic agent against cancer. PMID- 20013013 TI - Effects of different impact exercise modalities on bone mineral density in premenopausal women: a meta-analysis. AB - Our objective was to assess the effects of differing modes of impact exercise on bone density at the hip and spine in premenopausal women through systematic review and meta-analysis. Electronic databases, key journals and reference lists were searched for controlled trials investigating the effects of impact exercise interventions on lumbar spine (LS), femoral neck (FN) and total hip (TH) bone mineral density (BMD) in premenopausal women. Exercise protocols were categorised according to impact loading characteristics. Weighted mean difference (WMD) meta analyses were undertaken. Heterogeneity amongst trials was assessed. Fixed and random effects models were applied. Inspection of funnel plot symmetry was performed. Trial quality assessment was also undertaken. Combined protocols integrating odd- or high-impact exercise with high-magnitude loading (resistance exercises), were effective in increasing BMD at both LS and FN [WMD (fixed effect) 0.009 g cm(-2) 95% CI (0.002-0.015) and 0.007 g cm(-2) 95% CI (0.001 0.013); P = 0.011 and 0.017, respectively]. High-impact only protocols were effective on femoral neck BMD [WMD (fixed effect) 0.024 g cm(-2) 95% CI (0.002 0.027); P < 0.00001]. Funnel plots showed some asymmetry for positive BMD outcomes. Insufficient numbers of protocols assessing TH BMD were available for assessment. Exercise programmes that combine odd- or high-impact activity with high-magnitude resistance training appear effective in augmenting BMD in premenopausal women at the hip and spine. High-impact-alone protocols are effective only on hip BMD in this group. However, diverse methodological and reporting discrepancies are evident in published trials. PMID- 20013014 TI - Analysis of exon dosage using MLPA in South African Parkinson's disease patients. AB - Genomic rearrangements (exon dosage) are common mutations reported in Parkinson's disease (PD) patients. In the present study, we aimed to investigate the prevalence of genomic rearrangements in 88 South African patients with predominantly early-onset PD (age-at-onset 20 to 30 versus or=70 years versus <50 years, baseline corticosteroids, and persistent neutropenia. Survival was also significantly less likely for treatment in other regions versus North America and for patients with renal failure at baseline. These findings help to define non-antifungal drug factors that may impact survival and treatment success in invasive candidiasis or candidemia. PMID- 20013017 TI - The nodC, nodG, and glgX genes of Rhizobium tropici strain PRF 81. AB - Rhizobium tropici is a diazotrophic microsymbiont of common bean (Phaseolus vulgaris L.) that encompasses important but still poorly studied tropical strains, and a recent significant contribution to the knowledge of the species was the publication of a genomic draft of strain PRF 81, which revealed several novel genes [Pinto et al. Funct Int Gen 9:263-270, 2009]. In this study, we investigated the transcription of nodC, nodG, and glgX genes, located in the nod operon of PRF 81 strain, by reverse-transcription quantitative PCR. All three genes showed low levels of transcription when the cells were grown until exponential growth phase in the presence of common-bean-seed exudates or of the root nod-gene inducer naringenin. However, when cells at the exponential phase of growth were incubated with seed exudates, transcription occurred after only 5 min, and nodC, nodG, and glgX were transcribed 121.97-, 14.86-, and 50.29-fold more than the control, respectively, followed by a rapid decrease in gene transcription. Much lower levels of transcription were observed in the presence of naringenin; furthermore, maximum transcription required 8 h of incubation for all three genes. In light of these results, the mechanisms of induction of the nodulation genes by flavonoids are discussed. PMID- 20013018 TI - Stapled ileostomy closure results in reduction of postoperative morbidity. AB - BACKGROUND: Loop ileostomy is widely employed as a defunctioning procedure for left-sided colonic anastomoses. Closure of the stoma carries a risk of morbidity and even mortality. The aim of this prospective trial was to evaluate the ability of stapled stoma closure to decrease the rates of perioperative morbidity. METHODS: One hundred and nineteen patients (mean age 56.2 +/- 5.4 years) underwent two-stage operations for rectal carcinoma with protective loop ileostomy between 2005 and 2008. All patients were randomly divided into two groups: 56 patients had conventional ileostomy takedown, while in the other 63, a functional end-to-end anastomosis was created using a linear stapler. Groups were comparable in terms of age, gender, body mass index, and other parameters. RESULTS: Mean time of stoma closure using functional end-to-end anastomosis was 68 +/- 7, when compared to 92 +/- 11 min (P = 0.01) for conventional stoma closure. The overall morbidity rate after ileostomy closure using a stapler was 3.2%: one patient (1.6%) developed a wound infection and self-limited bleeding from the anastomotic line, while another patient (1.6%) had an ileal obstruction caused by adhesions and required additional intervention. Conventional ileostomy closure resulted in a 14.3% morbidity rate: six patients (10.7%) had prolonged ileus, 2 (3.6%) had small bowel obstruction, and 2 (3.6%) had wound infections (P = 0.04). CONCLUSION: Functional end-to-end anastomosis reduces operating time and morbidity compared to conventional ileostomy takedown. PMID- 20013019 TI - Physiological and chemical analysis of neurotransmitter candidates at a fast excitatory synapse in the jellyfish Cyanea capillata (Cnidaria, Scyphozoa). AB - Motor nerve net (MNN) neurons in the jellyfish Cyanea capillata communicate with one another by way of fast, bidirectional excitatory chemical synapses. As is the case with almost all identified chemical synapses in cnidarians, the identity of the neurotransmitter at these synapses is unclear. MNN neurons are large enough for stable intracellular recordings. This, together with the fact that they can be exposed, providing unlimited access to them and to their synapses, prompted a study of the action of a variety of neurotransmitter candidates, including those typically associated with fast synapses in higher animals. Only the amino acids taurine and beta-alanine produced physiological responses consistent with those of the normal EPSP in these cells. Moreover, chemical analysis revealed that both taurine and beta-alanine are present in the neurons and released by depolarization. These various findings strongly suggest that either or both of these amino acids, or a closely related compound is the neurotransmitter at the fast chemical synapses between MNN neurons. PMID- 20013020 TI - Effect of an electric field on the bifurcation of a biaxially stretched incompressible slab rubber. AB - This paper describes the effect of an electrical field on the bifurcation phenomenon that appears in a biaxially stretched slab of Mooney-Rivlin material (M. Mooney, J. Appl. Phys. 11, 582 (1940)) subjected to equal dead loads. The main conclusion of the analysis is that the stretch ratio at which the bifurcation phenomenon appears crucially depends on the configuration of the system rubber slab-electrodes. The theoretical foundations of the present study are based on a recent formulation on this subject carried out by Dorfmann and Ogden (A. Dorfmann, R.W. Ogden, Acta Mech. 174, 167 (2005); J. Elasticity 82, 99 (2006)). PMID- 20013021 TI - Developmental changes of the contingent negative variation in migraine and healthy children. AB - It has been hypothesized that abnormalities of information processing in migraine may be attributed to impairment of cerebral maturation. However, the most evidences for this hypothesis have come from cross-sectional studies during childhood. We performed a longitudinal study and recorded contingent negative variation (CNV), an event-related slow cortical potential, in migraine children (n = 27) and age-matched healthy individuals (n = 23) in 1998 and 8 years later (2006). Amplitudes of all CNV components were reduced and habituation of the initial CNV (iCNV) increased in the observed time. However, the reduction of the iCNV amplitude was more pronounced in migraine patients who were in remission in 2006 and in healthy subjects and less pronounced in migraineurs with persisting headaches. Patients with the worsened migraine demonstrated the most pronounced loss of iCNV habituation in 1998 and significantly increased iCNV amplitudes in 2006. This longitudinal study supports the hypothesis of impaired cerebral maturation in migraine and shows that migraine manifestation is a key factor interfering with the natural maturation process of central information processing. PMID- 20013022 TI - Functional interplay between E2F1 and chemotherapeutic drugs defines immediate E2F1 target genes crucial for cancer cell death. AB - The E2F1 transcription factor enhances apoptosis by DNA damage in tumors lacking p53. To elucidate the mechanism of a potential cooperation between E2F1 and chemotherapy, whole-genome microarrays of chemoresistant tumor cell lines were performed focusing on the identification of cooperation response genes (CRG). This gene class is defined by a synergistic expression response upon endogenous E2F1 activation and drug treatment. Cluster analysis revealed an expression pattern of CRGs similar to E2F1 mono-therapy, suggesting that chemotherapeutics enhance E2F1-dependent gene expression at the transcriptional level. Using this approach as a tool to explore E2F1-driven gene expression in response to anticancer drugs, we identified novel apoptosis genes such as the tumor suppressor TIEG1/KLF10 as direct E2F1 targets. We show that TIEG1/KLF10 is transcriptionally activated by E2F1 and crucial for E2F1-mediated chemosensitization of cancer cells. Our results provide a broader picture of E2F1 regulated genes in conjunction with cytotoxic treatment that allows the design of more rational therapeutics. PMID- 20013023 TI - Differential water permeability and regulation of three aquaporin 4 isoforms. AB - Aquaporin 4 (AQP4) is expressed in the perivascular glial endfeet and is an important pathway for water during formation and resolution of brain edema. In this study, we examined the functional properties and relative unit water permeability of three functional isoforms of AQP4 expressed in the brain (M1, M23, Mz). The M23 isoform gave rise to square arrays when expressed in Xenopus laevis oocytes. The relative unit water permeability differed significantly between the isoforms in the order of M1 > Mz > M23. None of the three isoforms were permeable to small osmolytes nor were they affected by changes in external K(+) concentration. Upon protein kinase C (PKC) activation, oocytes expressing the three isoforms demonstrated rapid reduction of water permeability, which correlated with AQP4 internalization. The M23 isoform was more sensitive to PKC regulation than the longer isoforms and was internalized significantly faster. Our results suggest a specific role for square array formation. PMID- 20013026 TI - Honorary memberships in the European Histamine Research Society. PMID- 20013025 TI - Hyaluronan reversed proteoglycan synthesis inhibited by mechanical stress: possible involvement of antioxidant effect. AB - INTRODUCTION: Abnormal mechanical stress loaded on the cartilage leads to the osteoarthritis (OA). Although intraarticular hyaluronan (HA) injection is an effective treatment for OA, the underlying mechanism has not been made clear. METHODS: Mechanical compression was loaded on the bovine cartilage using the Biopress system. Proteoglycan (PG) and reactive oxygen species (ROS) synthesis were measured with [(35)S] incorporation and fluorescent dye, respectively. Accumulation of peroxynitrite was determined with western blotting using nitrotyrosine antibody. RESULTS: Mechanical compression inhibited PG synthesis and enhanced ROS. Externally added HA reversed stress-inhibited PG synthesis and attenuated ROS synthesis. HA also significantly decreased the generation of nitrotyrosine. CONCLUSIONS: HA neutralized stress-enhanced ROS synthesis and resulted in the reversing of PG synthesis. These data suggest that HA plays an anabolic effect as an antioxidant. PMID- 20013024 TI - Platelets in defense against bacterial pathogens. AB - Platelets interact with bacterial pathogens through a wide array of cellular and molecular mechanisms. The consequences of this interaction may significantly influence the balance between infection and immunity. On the one hand, recent data indicate that certain bacteria may be capable of exploiting these interactions to gain a virulence advantage. Indeed, certain bacterial pathogens appear to have evolved specific ways in which to subvert activated platelets. Hence, it is conceivable that some bacterial pathogens exploit platelet responses. On the other hand, platelets are now known to possess unambiguous structures and functions of host defense effector cells. Recent discoveries emphasize critical features enabling such functions, including expression of toll like receptors that detect hallmark signals of bacterial infection, an array of microbicidal peptides, as well as other host defense molecules and functions. These concepts are consistent with increased risk and severity of bacterial infection as correlates of clinical abnormalities in platelet quantity and quality. In these respects, the molecular and cellular roles of platelets in host defense against bacterial pathogens are explored with attention on advances in platelet immunobiology. PMID- 20013027 TI - Effect of antioxidants on airway smooth muscle contraction: action of lipoic acid and some of its novel derivatives on guinea pig tracheal smooth muscle. AB - OBJECTIVES AND DESIGN: The aim of this study was to investigate the ability of (a) antioxidants, some related to alpha-lipoic acid (LA), (b) histone deacetylase (HDAC) inhibitors, and (c) hybrid compounds possessing both alpha-lipoic acid derived antioxidant properties and HDAC inhibitory activity to inhibit guinea pig smooth muscle contraction. MATERIALS AND METHODS: Guinea pig isolated tracheal rings (GPTR) were prepared and their isometric tension measured using a transducer. Histamine, carbachol and 5-hydroxytryptamine (5-HT) served as agonists. Tests with antigen (ovalbumin) used GPTR from sensitised guinea pigs or rings from non-sensitised animals that had been incubated for at least 2 h with diluted serum from sensitised animals. RESULTS: All antioxidants tested showed a relaxant effect on resting tension and on tension induced by histamine or carbachol, with EC(50)(s) of 0.2-5.0 mM and a rank order of potency: LA derivatives > glutathione (GSH) > ascorbic acid (AA). However, low concentrations (<50 microM) of GSH, AA and LA potentiated histamine-induced contractions. The benzamide HDAC inhibitor MGCD0103 inhibited mast cell activation and GPTR contraction produced by antigen and certain agonists, although a 2-6 h pre incubation was required for those effects to be apparent. Two LA-benzamide HDAC hybrid compounds, UCL M084 and UCL M109 inhibited GPTR contraction after 30 min pre-incubation; however, even after long pre-incubation (up to 6 h) those hybrid compounds showed less potent inhibition of agonist-induced contraction than did MGCD0103. CONCLUSIONS: The results showed that GSH more potently inhibited contraction induced by histamine than that induced by 5-HT or carbachol, whereas LA, and especially UCL M084 and UCL M109, more potently blocked contraction induced by carbachol and 5-HT than that induced by histamine. For GSH, and possibly also for LA-type compounds, the inhibition of agonist-induced tracheal smooth muscle contractions may be due to NO formation. This study did not detect a synergistic relaxant effect in two compounds incorporating the structural union of a benzamide HDAC inhibitor terminus with a LA-derived antioxidant moiety. PMID- 20013028 TI - Structural organization of mammalian copper-containing amine oxidase genes. AB - OBJECTIVE: Analysis of sequence conservation and structural organization of mammalian genes encoding copper-containing amine oxidases (CAO). METHODS: Sequences of previously characterized genes encoding CAO proteins were used to identify homologous mammalian genes in the NCBI genome sequence databases and to analyze sequence and structural conservation of these genes. RESULTS: Mammals possess four AOC genes encoding diamine oxidase (AOC1), retina-specific amine oxidase (AOC2), vascular adhesion protein-1 (AOC3), and serum amine oxidase (AOC4), with a defective AOC4 gene present in humans, mice, and rats. In addition to the common structure of all AOC genes, there is a high degree of interspecies sequence conservation for each of the four genes. CONCLUSIONS: Sequence and structural conservation of mammalian AOC genes implies a common evolutionary origin and functional diversification after gene duplication events. PMID- 20013029 TI - Use of lipid rafting for the analysis of human basophil activation by flow cytometry. AB - BACKGROUND: Human leukocyte activation induced by specific and non-specific stimuli is characterized by the formation of lipid rafts defined as lipid-ordered domains that are more tightly packed than the surrounding non-raft phase of the bilayer. These lipid rafts are formed in parallel with profound membrane reorganization. OBJECTIVES: Analyse the rafting and non-rafting proteins present on the activated and resting basophil membrane and study their interest for the flow cytometric analysis of basophil activation. METHODS: Human basophils obtained from samples used for diagnostic cellular tests such as basophil or lymphocyte activation tests were stimulated either by the formyl-methionyl-leucyl phenylalanine peptide (fMLP), by an anti-IgE or by an allergen. After 40 min at 37 degrees C, they were labelled by different antibodies conjugated to fluorescent dyes as an anti-IgE FITC, an anti-CCR3 PE, an anti-CD63, an anti CD203c PE, an anti-11b, annexin V FITC or by cholera toxin FITC. Moreover, several experiments were analysed using an Amnis cytometer, allowing one to obtain the picture of the analysed cells. RESULTS: Anti-IgE or specific allergen elicits a membrane neo expression of CD63 at a high density and is poorly represented on resting basophil membrane. Upon an IgE-dependant activation some of the markers already present on resting basophil membrane, as CD203c, are up regulated and others, such as the IgE/IgE FcepsilonRI receptor and CCR3 are down regulated and submitted to the formation of clusters demonstrated by the pictures taken with the Amnis cytometer. For non-IgE dependant activators, such as fMLP, the picture was different since IgE was not down regulated, whereas CCR3 was down regulated. As demonstrated using annexin V or the cholera toxin used for analysing apoptosis, these phenomenon were paralleled by the formation of lipid rafts, gangliosides domains, such as GM1, which is accessible from the extra cellular medium. CONCLUSIONS: Basophil activation leads to membrane events close to the apoptosis phenomenon. The flow cytometric analysis of these membrane events may lead to protocols for allergen-induced activation and, may significantly increase cellular test sensitivity, particularly for drugs allergy diagnosis for which the usual protocols, such as those using CD63 alone, are insufficiently sensitive. PMID- 20013030 TI - Luteolin inhibits cell proliferation during Azoxymethane-induced experimental colon carcinogenesis via Wnt/ beta-catenin pathway. AB - The protective role of Luteolin (LUT) against Azoxymethane (AOM)-induced mouse colon carcinogenesis has been documented earlier. The aim of this study is to investigate on the mechanism of chemopreventive action exhibited by LUT employing AOM-induced colon carcinogenesis in mice as an experimental model. LUT inhibited AOM-induced colon tumorigenesis by decreasing tumor incidence and size. LUT reduced the cell proliferation by decreasing the number of Argyrophillic nucleolar organizer region (AgNOR)/nucleus and Proliferating Cell Nuclear Antigen (PCNA) index. It was known that beta-catenin is a key effector in Wingless and Int (Wnt) signaling pathway and 90% of colon tumors arise from mutations in this pathway. In this study, we show evidence that LUT inhibited colon carcinogenesis by decreasing AOM-induced cell proliferation through the involvement of beta catenin, Glycogen synthase kinase (GSK)-3beta and cyclin D1, the key components in Wnt signaling pathway. In conclusion, the protective effect of LUT could be attributed to inhibition of AOM-induced cellular proliferation probably through the involvement of beta-catenin, GSK-3beta and cyclin D1. PMID- 20013033 TI - Pathways of metastasis suppression in bladder cancer. AB - Despite the recent advances in the diagnosis of bladder cancer, recurrence after surgical intervention for muscle invasive disease is still problematic as nearly half of the patients harbor occult distant metastases and this, in turn, is associated with poor 5-year survival rate. We have recently identified Rho family GDP dissociation inhibitor 2 (RhoGDI2) protein as functional metastasis suppressor and a prognostic marker in patients after cystectomy. In identifying the mechanisms underlying metastasis suppression by RhoGDI2, we found this protein to be associated with the c-Src kinase in human tumors, where the expression of both is diminished as a function of stage. Interestingly, c-Src bound to and phosphorylated RhoGDI2 resulting in enhanced metastasis suppressive potency. In this review, we will discuss the established roles of c-Src and RhoGDI2 in bladder cancer and speculate on their therapeutic relevance. PMID- 20013032 TI - Phosphatidylinositol 3-kinase (PI3K) pathway activation in bladder cancer. AB - The phosphatidylinositol 3-kinase (PI3K) pathway is a critical signal transduction pathway that regulates multiple cellular functions. Aberrant activation of this pathway has been identified in a wide range of cancers. Several pathway components including AKT, PI3K and mTOR represent potential therapeutic targets and many small molecule inhibitors are in development or early clinical trials. The complex regulation of the pathway, together with the multiple mechanisms by which it can be activated, make this a highly challenging pathway to target. For successful inhibition, detailed molecular information on individual tumours will be required and it is already clear that different tumour types show distinct combinations of alterations. Recent results have identified alterations in pathway components PIK3CA, PTEN, AKT1 and TSC1 in bladder cancer, some of which are significantly related to tumour phenotype and clinical behaviour. Co-existence of alterations to several PI3K pathway genes in some bladder tumours indicates that these proteins may have functions that are not related solely to the known canonical pathway. PMID- 20013034 TI - Urological cancers. Preface. PMID- 20013031 TI - Cross-species approaches to seed dormancy and germination: conservation and biodiversity of ABA-regulated mechanisms and the Brassicaceae DOG1 genes. AB - Seed dormancy is genetically determined with substantial environmental influence mediated, at least in part, by the plant hormone abscisic acid (ABA). The ABA related transcription factor ABI3/VP1 (ABA INSENSITIVE3/VIVIPAROUS1) is widespread among green plants. Alternative splicing of its transcripts appears to be involved in regulating seed dormancy, but the role of ABI3/VP1 goes beyond seeds and dormancy. In contrast, DOG1 (DELAY OF GERMINATION 1), a major quantitative trait gene more specifically involved in seed dormancy, was so far only known from Arabidopsis thaliana (AtDOG1) and whether it also has roles during the germination of non-dormant seeds was not known. Seed germination of Lepidium sativum ('garden cress') is controlled by ABA and its antagonists gibberellins and ethylene and involves the production of apoplastic hydroxyl radicals. We found orthologs of AtDOG1 in the Brassicaceae relatives L. sativum (LesaDOG1) and Brassica rapa (BrDOG1) and compared their gene structure and the sequences of their transcripts expressed in seeds. Tissue-specific analysis of LesaDOG1 transcript levels in L. sativum seeds showed that they are degraded upon imbibition in the radicle and the micropylar endosperm. ABA inhibits germination in that it delays radicle protrusion and endosperm weakening and it increased LesaDOG1 transcript levels during early germination due to enhanced transcription and/or inhibited degradation. A reduced decrease in LesaDOG1 transcript levels upon ABA treatment is evident in the late germination phase in both tissues. This temporal and ABA-related transcript expression pattern suggests a role for LesaDOG1 in the control of germination timing of non-dormant L. sativum seeds. The possible involvement of the ABA-related transcription factors ABI3 and ABI5 in the regulation of DOG1 transcript expression is discussed. Other species of the monophyletic genus Lepidium showed coat or embryo dormancy and are therefore highly suited for comparative seed biology. PMID- 20013035 TI - Parathyroid hormone PTH(1-34) formulation that enables uniform coating on a novel transdermal microprojection delivery system. AB - PURPOSE: Assess formulation parameters to enable >24-h continuous accurate and uniform coating of PTH(1-34) on a novel transdermal microprojection array delivery system. METHODS: Surface activity and rheology of the liquid formulation was determined by contact angle measurement and cone-plate viscometry. The formulation's delivery performance was assessed in vivo using the hairless guinea pig model. Peptide gelation was investigated by rheological and viscoelastic behavior changes. RESULTS: Accurate and uniform coating was achieved by formulating the liquid formulation to a preferred contact angle range of 30-60 degrees with a surfactant and by establishing a Newtonian fluid (defined as a fluid maintaining a constant viscosity with shear rate and time) with a viscosity of > or =20 cps via adjusting the peptide concentration and using an appropriate acidic counterion. A non-volatile acidic counterion was found critical to compensate for the loss of the volatile acetate counterion to maintain the peptide formulation's solubility upon rehydration in the skin. Finally, the 15.5% w/w PTH(1-34) concentration was found to be the most physically stable formulation (delayed gelation) in the roll-coating reservoir. With a properly designed coating reservoir for shear force reduction, the liquid formulation could last for more than 24 h without gelation. CONCLUSIONS: The study successfully offered scientific rationales for developing an optimal liquid formulation for a novel titanium microprojection array coating process. The resultant formulation has an enduring physical stability (>24 h) in the coating reservoir and maintained good in vivo dissolution performance. PMID- 20013036 TI - Bifunctional polymeric inhibitors of human influenza A viruses. AB - PURPOSE: New antiviral agents were prepared by attaching derivatives of sialic acid (1) and of the drug zanamivir (2) to poly(isobutylene-alt-maleic anhydride) (poly-(1 + 2)) or by mixing poly-1 and poly-2, followed by assaying them against wild-type and drug-resistant influenza A Wuhan viruses. METHODS: Individually or together, 1 and 2 were covalently bonded to the polymer. The antiviral potencies of the resultant poly-1, poly-2, poly-(1 + 2), and poly-1 + poly-2, as well as 1 and 2, were assessed using plaque reduction assay. RESULTS: Attaching 1 to the polymer improved at best millimolar IC(50) values over three orders of magnitude. While 2 exhibited micromolar IC(50) values, poly-2 was >100-fold even more potent. The IC(50) of poly-(1 + 2) against the wild-type strain was >300-fold and approximately 17-fold better than of poly-1 and poly-2, respectively. In contrast, the potency of poly-(1 + 2) vs. poly-2 against the mutant strain merely doubled. The mixture of poly-1 + poly-2 inhibited both viral strains similarly to poly-2. CONCLUSIONS: The bifunctional poly-(1 + 2) acts synergistically against the wild-type influenza virus, but not against its drug-resistant mutant, as compared to a physical mixture of the monofunctional poly-1 and poly-2. PMID- 20013037 TI - Deer responses to repellent stimuli. AB - Four repellents representing different modes of action (neophobia, irritation, conditioned aversion, and flavor modification) were tested with captive white tailed deer in a series of two-choice tests. Two diets differing significantly in energy content were employed in choice tests so that incentive to consume repellent-treated diets varied according to which diet was treated. When the high energy diet was treated with repellents, only blood (flavor modification) and capsaicin (irritation) proved highly effective. Rapid habituation to the odor of meat and bone meal (neophobia) presented in a sachet limited its effectiveness as a repellent under conditions with a high feeding motivation. Thiram, a stimulus used to condition aversions, was not strongly avoided in these trials, that included only limited exposures to the repellent. These data support previous studies indicating that habituation to odor limits the effectiveness of repellents that are not applied directly to food, while topically-applied irritants and animal-based products produce significant avoidance. PMID- 20013038 TI - Hydrocarbon footprints as a record of bumblebee flower visitation. AB - Bumblebees leave traces of cuticular hydrocarbons on flowers they visit, with the amount deposited being positively related to the number of visits. We asked whether such footprint hydrocarbons are retained on flowers for sufficiently long periods of time so as to reflect bee visitation in pollination studies. In laboratory experiments, flower corollae (Primula veris, Digitalis grandiflora) visited by Bombus terrestris workers retained bee-derived nonacosenes (C(29)H(58)) in near-unchanged quantities for 24 hours, both at 15 and 25 degrees C. Additionally, synthetic (Z)-9-tricosene applied to flower corollae of the deadnettle Lamium maculatum was retained for 48 hours in an unchanged quantity. In a field survey, the amount of footprint alkenes on flowers of comfrey (Symphytum officinale) plants was positively correlated with the number of bumblebee visits that those plants had received during the day. Together, these data suggest that flowers retain a long-term quantitative record of bumblebee visitation. The analysis of petal extracts by gas chromatography could provide a cheap and reliable way of quantifying bumblebee visits in landscape scale studies of pollination. PMID- 20013039 TI - Emission of volatile organic compounds after herbivory from Trifolium pratense (L.) under laboratory and field conditions. AB - Plants emit a wide range of volatile organic compounds in response to damage by herbivores, and many of the compounds have been shown to attract the natural enemies of insect herbivores or serve for inter- and intra-plant communication. Most studies have focused on volatile emission in the laboratory while little is known about emission patterns in the field. We studied the emission of volatiles by Trifolium pratense (red clover) under both laboratory and field conditions. The emission of 24 compounds was quantified in the laboratory, of which eight showed increased emission rates after herbivory by Spodoptera littoralis caterpillars, including (E)-beta-ocimene, the most abundant compound, (Z)-beta ocimene, linalool, (E)-beta-caryophyllene, (E,E)-alpha-farnesene, 4,8-dimethyl 1,3,7-nonatriene (DMNT), 1-octen-3-ol, and methyl salicylate (MeSA). While most of these compounds have been reported as herbivore-induced volatiles from a wide range of plant taxa, 1-octen-3-ol seems to be a characteristic volatile of legumes. In the field, T. pratense plants with varying herbivore damage growing in established grassland communities emitted only 13 detectable compounds, and the correlation between herbivore damage and volatile release was more variable than in the laboratory. For example, the emission of (E)-beta-ocimene, (Z)-beta ocimene, and DMNT actually declined with damage, while decanal exhibited increased emission with increasing herbivory. Elevated light and temperature increased the emission of many compounds, but the differences in light and temperature conditions between the laboratory and the field could not account for the differences in emission profiles. Our results indicate that the release of volatiles from T. pratense plants in the field is likely to be influenced by additional biotic and abiotic factors not measured in this study. The elucidation of these factors may be important in understanding the physiological and ecological functions of volatiles in plants. PMID- 20013040 TI - Capnography to check the safety of Bain circuit. AB - The test of the inner tube integrity is an important checklist prior to the safe use of Bain's breathing system. This is because the major concern with the use of Bain's circuit is the potential malfunctioning of the circuit due to avulsion of the inner fresh gas delivery tube at the machine end which will turn the outer tube into dead space, a hazard recognized by Hannallah. Pethick test although widely used may not be foolproof to detect leaks in the inner tube. Ghani suggested the use of a plunger to perform the inner tube occlusion. Partially or completely occluded outer tube may produce a false positive result. Using very high flows or prolonged occlusion may cause damage to anesthetic machine due to high pressure. PMID- 20013041 TI - Personal pronouns and communicative engagement in autism. AB - In three experimental conditions, we tested matched children with and without autism (n = 15 per group) for their comprehension and use of first person plural ('we') and third person singular ('he') pronouns, and examined whether such linguistic functioning related to their social interaction. The groups were indistinguishable in their comprehension and use of 'we' pronouns, although within each group, such usage was correlated with ratings of interpersonal connectedness with the collaborator. On the other hand, participants with autism were less likely to use third person pronouns or to show patterns of eye gaze reflecting engagement with an interlocutor's stance vis-a-vis a third person. In these settings, atypical third person pronoun usage seemed to reflect limited communicative engagement, but first person pronouns were relatively spared. PMID- 20013042 TI - Alcohol consumption is associated with poor health in HIV clinic patient population: a behavioral surveillance study. AB - Previous research reports that populations with HIV consume higher rates of alcohol than general population. This cross-sectional study (n = 391) was conducted to measure alcohol consumption, factors associated with consumption, and the relationship between alcohol and HIV viral loads among individuals receiving HIV care. Increased alcohol consumption was associated with being male, lower education attainment, and lacking a current HAART prescription. Additionally, among those currently on HAART, unsuppressed viremia was associated with higher levels of alcohol consumption. These findings suggest that there may be a relatively low level of alcohol consumption that is detrimental to virologic suppression among populations with HIV. PMID- 20013044 TI - Viability of using employment rates from randomized trials as benchmarks for supported employment program performance. AB - Cumulative employment rates published by randomized trials are based on each enrollee's pre-planned 18-24-months of study participation. By contrast, community programs typically report employment rates for clients active in services during a calendar quarter. Using data from three supported employment programs in randomized trials, we show that trial cumulative employment rates are about twice as large as quarterly employment rates for the same program. Therefore, we recommend that administrators, service networks, and mental health authorities begin to publish quarterly employment rates, and quarterly median earnings, to allow policymakers to set realistic performance expectations for supported employment programs. PMID- 20013045 TI - Patterns of physical and relational aggression in a school-based sample of boys and girls. AB - The current study investigated the patterns of aggressive behavior displayed in a sample of 282 students in the 4th through 7th grades (M age = 11.28; SD = 1.82). Using cluster analyses, two distinct patterns of physical aggression emerged for both boys and girls with one aggressive cluster showing mild levels of reactive aggression and one group showing high levels of both reactive and proactive aggression. Both aggressive clusters showed problems with anger dysregulation, impulsivity, thrill and adventure seeking, positive outcome expectancies for aggression, and higher rates of bullying. However, the combined cluster was most severe on all of these variables and only the combined aggressive group differed from non-aggressive students on their level of callous-unemotional traits. Similar patterns of findings emerged for relational aggression but only for girls. PMID- 20013043 TI - Mental health treatment to reduce HIV transmission risk behavior: a positive prevention model. AB - Secondary HIV prevention, or "positive prevention," is concerned with reducing HIV transmission risk behavior and optimizing the health and quality of life of people living with HIV/AIDS (PLWHA). The association between mental health and HIV transmission risk (i.e., sexual risk and poor medication adherence) is well established, although most of this evidence is observational. Further, a number of efficacious mental health treatments are available for PLWHA yet few positive prevention interventions integrate mental health treatment. We propose that mental health treatment, including behavioral and pharmacologic interventions, can lead to reductions in HIV transmission risk behavior and should be a core component of secondary HIV prevention. We present a conceptual model and recommendations to guide future research on the effect of mental health treatment on HIV transmission risk behavior among PLWHA. PMID- 20013047 TI - No association between CYP17 T-34C polymorphism and breast cancer risk: a meta analysis involving 58,814 subjects. AB - Breast cancer is one of the most common malignant tumors worldwide. To date, many articles have evaluated the association between Cytochrome P450c17 (CYP17) T-34C polymorphism and breast cancer risk. However, the results remain inconclusive. In order to derive a more precise estimation of the association, a meta-analysis was performed in this study. By searching Medline, ISI Web of Knowledge, Cochrane, ScienceDirect, EBSCO, CNKI, and SinoMed databases, 43 studies including 26,008 cases and 32,806 controls were collected for CYP17 T-34C polymorphism. Crude ORs with 95% CIs were used to assess the strength of association between CYP17 T-34C polymorphism and breast cancer risk. The pooled ORs were performed for codominant model, dominant model, and recessive model, respectively. Overall, no significant associations between CYP17 T-34C polymorphism and breast cancer susceptibility were found for TT versus CC (OR = 0.96; 95% CI: 0.89-1.05), TC versus CC (OR = 0.97; 95% CI: 0.89-1.06), TT + TC versus CC (OR = 0.97; 95% CI: 0.89-1.05) and TT versus TC + CC (OR = 0.98; 95% CI: 0.93-1.03). In the stratified analysis by ethnicity, menopausal status, and sources of controls, significant associations were still not detected in all genetic models. In conclusion, this meta-analysis strongly suggests that CYP17 T-34C polymorphism is not associated with breast cancer risk. PMID- 20013046 TI - The relations among measurements of informant discrepancies within a multisite trial of treatments for childhood social phobia. AB - Discrepancies between informants' reports of children's behavior are robustly observed in clinical child research and have important implications for interpreting the outcomes of controlled treatment trials. However, little is known about the basic psychometric properties of these discrepancies. This study examined the relation between parent-child reporting discrepancies on measures of child social phobia symptoms, administered before and after treatment for social phobia. Participants included a clinic sample of 81 children (7-16 years old [M = 11.75, SD = 2.57]; 39 girls, 42 boys) and their parents receiving treatment as part of a multisite controlled trial. Pretreatment parent-child reporting discrepancies predicted parent-child discrepancies at posttreatment, and these relations were not better accounted for by the severity of the child's pretreatment primary diagnosis. Further, treatment responder status moderated this relation: Significant relations were identified for treatment non-responders and not for treatment responders. Overall, findings suggest that informant discrepancies can be reliably employed to measure individual differences over the course of controlled treatment trials. These data provide additional empirical support for recent work suggesting that informant discrepancies can meaningfully inform understanding of treatment response as well as variability in treatment outcomes. PMID- 20013048 TI - Can juvenile corals be surveyed effectively using digital photography?: implications for rapid assessment techniques. AB - The widespread decline of coral reefs requires integrated management measures across whole regions. Knowledge of demographic processes of reef organisms is important for informed management, yet current techniques for assessing such processes are time consuming, making it impractical to gather relevant information over large scales. We tested the usefulness of digital still photography as a rapid assessment technique to estimate coral recruitment--an important process in coral reef recovery. Estimates of the density and diversity of juvenile hard corals from digital images were compared with direct visual estimates from the same plots made in the field. Multiple plots were sampled on four reefs from a range of locations on Australia's Great Barrier Reef. On average, estimates of juvenile densities from photographic images were lower, in both absolute and relative terms, than that estimated from images. This was the case whether colonies <20 mm or <50 mm in diameter were considered. Overall differences between methods were generally greater at reefs where recruitment was higher, though proportional differences (density from images/density from direct visual census) still varied among reefs. Although the ranking of taxa, in terms of their densities, from the two methods were similar, the density of common genera was generally underestimated in images, and the occurrence of 'unknown' taxa was higher. We conclude that photographic images do not constitute a reliable rapid assessment method for estimating the spatial patterns in the density or diversity of juvenile hard corals. PMID- 20013049 TI - Identification of organic phosphorus compounds in the Bronx River bed sediments by phosphorus-31 nuclear magnetic resonance spectroscopy. AB - Sediment characteristics influence the distribution and bioavailability of phosphorus (P) in rivers and lakes. The objectives of this study were to identify P compounds in sediments collected from 15 sites along the Bronx River to get insights on nutrient transport for management of highly variable and modified ecosystems such as the Bronx River. The nuclear magnetic resonance spectra showed that the dominant P species in Bronx River bed sediments are orthophosphate monoester and lesser phosphate diesters and pyrophosphates (pyro-P). The P compounds were mostly glycerophosphate, nucleoside monophosphates, and polynucleotides. A few sites showed a small amount of dihydroxyacetone phosphate, inosine monophosphate. By allowing a downstream comparison of P compound variations along the Bronx River, this study provides a step toward improving water quality in an urban river system such as New York City and helps to assess the bioavailability of P, in turn, design estuary habitat restoration projects in comparable region of the world. PMID- 20013050 TI - A new PCR-RFLP method for detection of Anaplasma marginale based on 16S rRNA. AB - Species of the genus Anaplasma (Rickettsiales: Anaplasmataceae) are obligate intracellular tick borne pathogens. Three species of Anaplasma that infect cattle and sheep (A. marginale, A. centrale and A. ovis) are well recognized. Of these erythrocytic Anaplasma, A. marginale can cause diseases in the livestock with high economical losses. Species-specific PCR based on 16S rRNA gene is commonly used for detection of Anaplasma species but can not differentiate A. marginale, A. centrale and A. ovis because of sequence similarity. In this study DNA extraction was performed on 50 blood samples with presence of Anaplasma spp. in marginal point of erythrocytes in their blood smears. The extracted DNA from blood cells was analyzed by PCR and PCR-RFLP using primers derived from 16S rRNA gene and restriction endonuclease Bst1107 I. The restriction endonuclease Bst1107I only recognizes the sequence (GTATAC) in corresponding PCR product of A. marginale and cut it. The nucleotide sequence of the A. marginale 16S rRNA gene was determined and compared with the sequences of A. marginale in GenBank. The 16S rRNA of A. marginale in Iran was completely similar to the related sequence deposited in GenBank at accession number of M60313. In the present study we propose a new PCR-restriction fragment length polymorphism analysis (RFLP) method based on 16S rRNA gene for specific detection of A. marginale. PMID- 20013051 TI - Silver ion treatment of primary cultured bovine mammary gland epithelial cell (BMEC) damage from Staphylococcus aureus-derived alpha-toxin. AB - Bovine mastitis remains the largest hazard in the global dairy industry and has facilitated the development of various therapeutic strategies. Silver is a well known disinfectant that is widely used in the treatment of clinical disease. In this study, we separated bovine mammary gland epithelial cells (BMEC) using an enzyme probe. We also examined safe concentrations for the application of silver ions in bovine mastitis, particularly in cases induced by Staphylococcus aureus. S. aureus-derived alpha-toxins induced cell damage through DNA fragmentation, reactive oxygen species (ROS) generation, and the dissipation of mitochondrial transmembrane potential (MTP) in BMEC. Silver ion treatment doses of lower than 2 ppm did not induce BMEC damage, but silver ion concentrations greater than 4 ppm was accompanied by DNA fragmentation. Furthermore, silver ions doses below 2 ppm inhibited alpha-toxin-induced cell damage through the reduction of ROS generation. Recognizing this, it demonstrate that low doses of silver ions inhibit alpha-toxin-induced BMEC damage and suggest that silver ions may be a potentially beneficial treatment against bovine mastitis, particularly in cases induced by S. aureus. PMID- 20013053 TI - The use of seromuscular tapered ileal tube in ureteral replacement: an experimental model. AB - PURPOSE: To assess the capability of urothelium to proliferate, creep and line the inner surface of the interposed seromuscular tapered ileal tube. MATERIALS AND METHODS: Under general anesthesia, 15 female dogs underwent resection of 5 cm of the mid ureter and replaced with tapered seromuscular ileal tube stented for 6 weeks. The animals were sacrificed, and cross section of the ileal ureters were examined histologically for the lining cells. RESULTS: Multilayer of transitional epithelium was seen covering all the inner surface of the interposed seromuscular tube at the end of 6 weeks. Excessive inflammatory cell infiltration was a prominent finding in the submucosal layer. CONCLUSION: Transitional epithelium has the capability to proliferate, grow and cover the inner surface of the interposed seromuscular ileal tube. Urothelium lining avoid the metabolic complications of the intestinal mucosa. PMID- 20013054 TI - Hyponatremia associated with paroxetine induced by sodium-restricted diet and hypotonic saline. AB - CASE DESCRIPTION: An 89-year-old woman taking paroxetine was admitted to our hospital for femoral neck fracture; her diet became sodium restricted due to hypertension. After admission, the femoral head replacement was performed and hypotonic saline was administered over 2 days. On the fifth day after the operation, severe hyponatremia was observed and treated with oral fluid restriction, furosemide, sodium chloride and paroxetine discontinuance. In a few days, serum sodium concentration returned to baseline level. CONCLUSIONS: Besides risk factors for SIADH, a sodium-restricted diet and infusions of hypotonic saline in the perioperative period should be considered risk factors for SIADH associated with paroxetine. PMID- 20013055 TI - Chlorella pyrenoidosa supplementation reduces the risk of anemia, proteinuria and edema in pregnant women. AB - Pregnancy anemia and pregnancy-induced hypertension (PIH) are common and potentially dangerous disorder in human pregnancy, and nutritional status of pregnant women is one of the leading causes. Chlorella contains large quantities of folate, vitamin B-12 and iron, and can help improve anemia and hypertensive disorder. Our objective was to investigate the preventive effects of Chlorella supplement on pregnancy anemia and PIH in Japanese pregnant women. A total of 70 pregnant women were placed into the control group (n = 38) or the Chlorella group (n = 32). The subjects in the Chlorella group were supplemented daily from 12th 18th wk of gestation until delivery with 6 g of Chlorella supplement. The proportion of anemic (hemoglobin level < 11 g/dL) subjects in the Chlorella group were significantly lower compared with the control group at the second and third trimesters. Additionally, in the Chlorella group, the incidences of proteinuria and edema, signs of PIH, were significantly lower during the third trimester. These results suggest that Chlorella supplementation significantly reduces the risk of pregnancy associated anemia, proteinuria and edema. Chlorella supplement may be useful as a resource of natural folate, vitamin B-12 and iron for pregnant women. PMID- 20013056 TI - Proanthocyanidins in buckwheat flour can reduce salivary nitrite to nitric oxide in the stomach. AB - Buckwheat flour, which is used for various dishes in the world, is a good source of proanthocyanidins. Proanthocyanidins in the buckwheat flour reduced nitrous acid producing nitric oxide (NO) when the flour was suspended in acidified saliva or in acidic buffer solution in the presence of nitrite. The ingestion of dough prepared from buckwheat flour increased the concentration of NO in the air expelled from the stomach, suggesting that the proanthocyanidins also reduced nitrite to NO in the stomach. During the production of NO by the buckwheat flour/nitrous acid systems, oxidation, nitration, and nitrosation of proanthocyanidins proceeded. The increase in the concentration of NO could improve the activity of stomach helping the digestion of ingested foods and the nitration and nitrosation of the proanthocyanidins could contribute to the scavenging of reactive nitrogen oxide species generated from NO and nitrous acid. PMID- 20013057 TI - cDNA cloning and expression analysis of wheat (Triticum aestivum L.) phytoene and zeta-carotene desaturase genes. AB - Carotene desaturation, an essential step in the carotenoid biosynthesis pathway, is catalyzed by two carotene desaturases, phytoene desaturase (PDS) and zeta carotene desaturase (zeta-carotene desaturase, ZDS). Full-length cDNAs designated TaPDS and TaZDS were cloned from wheat (Triticum aestivum cv. Chinese Spring) respectively, using the rapid amplification of cDNA ends (RACE) approach. The cDNA of TaPDS sequence was 2076 bp long, containing a 1731 bp open reading frame (ORF) which deduced protein having 576 amino acid residues with predicted molecular mass of 64.3 kDa and having a putative transit sequence for plastid targeting in the N-terminal region. While the cDNA sequence of TaZDS was 2150 bp long, contained an ORF sequence of 1707 bp, and encoded a putative protein of 568 amino acid residues with an estimated molecular mass of 62.5 kDa. Phylogenetic analysis demonstrated that TaPDS and TaZDS showed high homology with other PDSs and ZDSs in higher plant species, respectively. Moreover, sequences analysis also showed a high degree of conservation among plant PDSs and ZDSs. The deduced TaPDS and TaZDS protein both have the dinucleotide binding domain and conserved regions characteristic of other carotene desaturases. Analysis of the expression pattern of wheat TaPDS and TaZDS in different tissues revealed that the transcripts levels were higher in leaves and flowers petals, followed by in inflorescences, and were nearly absent in the roots and seeds. Southern analysis of genomic DNA indicated that the wheat TaPDS and TaZDS probably belong to a low-copy-number gene family. PMID- 20013058 TI - Relationship between methods of coping, social support and receipt of preventive care procedures by primary grandmother caregivers. AB - The purpose of this study was to examine the relationship between receipt of preventive care procedures, social support and methods used by primary grandmother caregivers to cope with the demands of caregiving. A self administered questionnaire comprised of items on preventive procedures and standardized measures of social support and methods of coping was used to collect data from a sample of 86 primary grandmother caregivers (56 +/- 8 years). Results show that blood pressure check ups (94%) and general physical examination (89%) were the preventive care procedures received by most participants. The average adherence score for the nine preventive care procedures was 6.60 +/- 2.10 and 50% of the participants had received each of the nine preventive care procedures considered by this study. Significant relationships were found between avoidance coping and receiving a general physical examination and eye or vision examination; between coping by minimizing the situation and receiving eye or vision examination; and between active coping and receiving a cholesterol check. Subjective and instrumental social supports were both significantly associated with receiving a gynecological examination or Pap smear and blood pressure check up. In conclusion, findings of this study show that primary grandmother caregivers have a high propensity towards preventive care. However healthcare providers providing preventive care to primary grandmother caregivers and other informal caregivers should assess the social supports and methods of coping used to cope with the demands of caregiving because both affect receipt of preventive care. PMID- 20013059 TI - Dental services utilization by women of childbearing age by socioeconomic status. AB - For women of childbearing age, oral health not only affects their physical and psychological well-being but also that of their children. This study used the 2003-2004 Ohio Family Health Survey (N = 9,819) to examine dental need and utilization by women in Ohio. Predisposing, enabling, and need variables were examined as they effect dental health service utilization by women of childbearing age at different socioeconomic status (SES) levels. The proportion of women in the low SES group self reporting a dental need (18%) was 3 times that of the proportion of women in the higher SES group with a self reported need (6%). Results of bivariate analysis showed that having a dental visit in the past year varied significantly by SES, race, insurance status, provider density, and need. A racial disparity in dental service utilization was noted in the bivariate analysis of the middle SES group. While dental need and type of dental coverage varied by SES, both were significantly associated with utilization of dental services within all 3 SES categories in the logistic regressions. These results suggest that measures need to be implemented to meet the goal of increasing access and utilization of dental health services by low-income populations. PMID- 20013060 TI - Conducting health research in Korean American churches: perspectives from church leaders. AB - Korean Americans experience many challenges to obtaining adequate health care coverage and access to needed services. Because a large proportion of Korean Americans attend churches on a regular basis, churches may be a promising venue where health programs can be delivered. In order to gain an in-depth understanding of Korean American churches with respect to conducting future health intervention research, we conducted exploratory interviews and focus groups with 58 leaders from 23 Korean American churches and three community organizations. From these interviews and focus groups, we found that Korean churches and church leaders seek to meet a variety of social and health needs of their congregation and their surrounding community. Several leaders have stated that assisting with social and medical needs of their members is an important component of their current ministry. They described profound health needs of their congregations and have suggested various ways in which the university can partner with the local churches to help address these needs through research. Additionally, they described various resources churches can provide to researchers such as: their personal assistance, church volunteer base, church facility, and church network and contacts. Our findings suggest that Korean churches have a high potential to serve an important role in the health of Korean Americans. On the basis of the promising results of the present study, we are planning to conduct a cross sectional survey of Korean church leaders and members in Los Angeles County to substantiate our findings in a larger representative sample. PMID- 20013061 TI - Factors associated with ethnic differences in health insurance coverage and type among Asian Americans. AB - This study examines the discrepancies in health insurance coverage and type across Asian American ethnic groups and the potential factors that may explain why these differences exist. Asian Americans are often considered as a homogeneous population and consequently, remain largely "invisible" in the current research literature. Recent data have highlighted discrepancies in the health insurance coverage between different Asian American ethnic groups particularly the high uninsurance rates among Korean and Vietnamese Americans. For this study, the 2003 and 2005 California Health Interview Surveys were pooled to obtain a sample of 6,610 Asian American adults aged 18-64, including those of Chinese, Filipino, Japanese, South Asian, and Vietnamese ethnicity. Binomial and multinomial logistic regression models were used to examine the likelihood of current health coverage and insurance type (employer-based vs. private vs. public), respectively. The results showed that ethnic differences in uninsurance and insurance type were partially explained by socioeconomic and immigration related characteristics-particularly for Vietnamese Americans and to a lesser extent, for Chinese and Korean Americans. There were also key differences in the extent to which specific ethnic groups purchased private insurance or relied on public programs (e.g., Medicaid) to offset the lack of employer-based coverage. This study reaffirms the tremendous heterogeneity in the Asian American population and the need for more targeted policy approaches. With the lack of adequate national data, more localized studies may help to improve our understanding of the health issues affecting specific Asian ethnic groups. PMID- 20013062 TI - Using the theory of planned behavior to explain the drinking motivations of social, high-risk, and extreme drinkers on game day. AB - This study assessed the extent to which the Theory of Planned Behavior (TPB) correctly predicted college student's motivation to consume alcohol on game day based on alcohol consumption rates. Three cohorts of 1,000 participants each (N = 3,000) were randomly selected and invited to complete an anonymous web-based survey the Monday following one of three designated college home football games. Path analyses were conducted to determine which of the TPB constructs were most effective in predicting Behavioral Intention and alcohol consumption among social, high-risk, and extreme drinkers. Social drinkers, high-risk, and those drinkers who engage in Extreme Ritualistic Alcohol Consumption (ERAC) were defined as males who consumed 1-4, 5-9, or 10 or more drinks on game day (1-3, 4 8, or nine or more drinks for females), respectively. Attitude Towards the Behavior and Subjective Norm constructs predicted participant's intentions to consume alcohol and corresponding behavior among all three classifications of drinkers; whereas the Perceived Behavioral Control (PBC) construct inconsistently predicted intention and alcohol consumption. Based on Behavioral Intention, the proportion of variance the TPB model explained decreased as participants alcohol consumption increased. It appears that the TPB constructs Attitude Toward the Behavior and Subjective Norm can effectively be utilized when designing universal prevention interventions targeting game day alcohol consumption among college students. However, the applicability of the PBC construct remains in question. While select constructs in the TPB appear to have predictive ability, the usefulness of the complete theoretical framework is limited when trying to predict high-risk drinking and ERAC. These findings suggest that other behavioral theories should be considered when addressing the needs of high-risk and extreme drinkers. PMID- 20013063 TI - pcDNA3.1tdTomato is superior to pDsRed2-N1 for optical fluorescence imaging in the F344/AY-27 rat model of bladder cancer. AB - PURPOSE: Animal models are important for pre-clinical assessment of novel therapies in metastatic bladder cancer. The F344/AY-27 model involves orthotopic colonisation with AY-27 tumour cells which are syngeneic to F344 rats. One disadvantage of the model is the unknown status of colonisation between instillation and sacrifice. Non-invasive optical imaging using red fluorescence reporters could potentially detect tumours in situ and would also reduce the number of animals required for each experiment. MATERIALS AND METHODS: AY-27 cells were stably transfected with either pDsRed2-N1 or pcDNA3.1tdTomato. The intensity and stability of fluorescence in the resultant AY-27/DsRed2-N1 and AY 27/tdTomato stable cell lines were compared using Xenogen IVIS(r)200 and Olympus IX51 systems. RESULTS: AY-27/tdTomato fluorescence intensity was 60-fold brighter than AY-27/DsRed2-N1 and was sustained in AY-27/tdTomato cells following freezing and six subsequent sub-cultures. After sub-cutaneous injection, fluorescence intensity from AY-27/tdTomato cells was threefold stronger than that detected from AY-27/DsRed2-N1 cells. IVIS(r)200 detected fluorescence from AY-27/tdTomato and AY-27/DsRed2-N1 cells colonising resected and exteriorised bladders, respectively. However, the deep-seated position of the bladder precluded in vivo imaging. Characteristics of AY-27/tdTomato cells in vitro and in tumours colonising F344 rats resembled those of parental AY-27 cells. Tumour transformation was observed in the bladders colonised with AY-27/DsRed2-N1 cells. CONCLUSIONS: In vivo whole-body imaging of internal red fluorescent animal tumours should use pcDNA3.1tdTomato rather than pDsRed2-N1. Optical imaging of deep-seated organs in larger animals remains a challenge which may require proteins with brighter red or far-red fluorescence and/or alternative approaches. PMID- 20013064 TI - Antioxidants improve early survival of cardiomyoblasts after transplantation to the myocardium. AB - PURPOSE: We tested the hypothesis that modulation of the microenvironment (using antioxidants) will increase stem cell survival in hypoxia and after transplantation to the myocardium. PROCEDURES: Rat cardiomyoblasts were stably transfected with a reporter gene (firefly luciferase) for bioluminescence imaging (BLI). First, we examined the role of oxidative stress in cells under hypoxic conditions. Subsequently, stem cells were transplanted to the myocardium of rats using high-resolution ultrasound, and their survival was monitored daily using BLI. RESULTS: Under hypoxia, oxidative stress was increased together with decreased cell survival compared to control cells, both of which were preserved by antioxidants. In living subjects, oxidative stress blockade increased early cell survival after transplantation to the myocardium, compared to untreated cells/animals. CONCLUSION: Modulation of the local microenvironment (with antioxidants) improves stem cell survival. Increased understanding of the interaction between stem cells and their microenvironment will be critical to advance the field of regenerative medicine. PMID- 20013065 TI - Comments on pesticide risk assessment by the revision of Directive EU 91/414. AB - INTRODUCTION: Human health and the environment are major concerns for European Commission policy on the authorisation of plant protection products. The new regulation that revises and replaces the directive 91/414/EC moves towards the adoption of a Persistent Bioaccumulation Toxicity cutoff criterion because current pesticide risk assessment (PRA) is deterministic, based on few standard cases and therefore characterised by uncertainty. This revision could create concerns about sustainability. DISCUSSION: This paper analyses some effects of this directive on the agrochemical market and assumes new effects resulting from the introduction of the revision. Suggestions are made as to how pesticide risk assessment will have to adapt to answer the request of legislators on safety standards and sustainability, introducing probabilistic PRA. Toxicity and exposure functions will be fully characterised, producing distributions of predicted impact and quantifying the variability and uncertainty. For adopting PRA studies at the local/catchment scale, new assessment schemes will be necessary. PMID- 20013066 TI - Burnout in the mental health workforce: a review. AB - There are enormous concerns regarding the recruitment, retention, training, and performance of the behavioral health workforce. Paramount among these concerns is turnover, which causes disruption in continuity of care, diminishes access to care while a position remains vacant, and poses financial hardship on the provider organization through costs related to recruitment, orientation, and training of a new hire. There is frequent mention of burnout within the literature and among behavioral health managers as a potential cause of turnover. However, there is no recent or comprehensive review of burnout that examines the evidence surrounding its validity, utility, and potential relationship to turnover. The purpose of this paper is to provide such a review by examining the construct of burnout, methodological and measurement issues, its prevalence in the mental health workforce, correlates of burnout, and interventions to decrease it. The implications for provider organizations and recommendations for future research are identified. PMID- 20013067 TI - Idiopathic granulomatous mastitis: case reports and review of literature. AB - Idiopathic granulomatous mastitis (IGM) is an uncommon benign disorder of the breast that can mimic two frequent breast disorders, breast carcinoma and breast abscess. In this report, we present two patients seen in a community teaching hospital over a period of one year, diagnosed with IGM after histological evaluation. One patient responded well to immunosuppressive therapy, but the second patient required bilateral mastectomy due to the severe and recurrent nature of the disease. IGM is a disorder that should be considered in the evaluation of women who present with painful breast disease. We discuss the diagnosis, clinical presentation and management of IGM. PMID- 20013068 TI - Hospital readmission in general medicine patients: a prediction model. AB - BACKGROUND: Previous studies of hospital readmission have focused on specific conditions or populations and generated complex prediction models. OBJECTIVE: To identify predictors of early hospital readmission in a diverse patient population and derive and validate a simple model for identifying patients at high readmission risk. DESIGN: Prospective observational cohort study. PATIENTS: Participants encompassed 10,946 patients discharged home from general medicine services at six academic medical centers and were randomly divided into derivation (n = 7,287) and validation (n = 3,659) cohorts. MEASUREMENTS: We identified readmissions from administrative data and 30-day post-discharge telephone follow-up. Patient-level factors were grouped into four categories: sociodemographic factors, social support, health condition, and healthcare utilization. We performed logistic regression analysis to identify significant predictors of unplanned readmission within 30 days of discharge and developed a scoring system for estimating readmission risk. RESULTS: Approximately 17.5% of patients were readmitted in each cohort. Among patients in the derivation cohort, seven factors emerged as significant predictors of early readmission: insurance status, marital status, having a regular physician, Charlson comorbidity index, SF12 physical component score, >or=1 admission(s) within the last year, and current length of stay >2 days. A cumulative risk score of >or=25 points identified 5% of patients with a readmission risk of approximately 30% in each cohort. Model discrimination was fair with a c-statistic of 0.65 and 0.61 for the derivation and validation cohorts, respectively. CONCLUSIONS: Select patient characteristics easily available shortly after admission can be used to identify a subset of patients at elevated risk of early readmission. This information may guide the efficient use of interventions to prevent readmission. PMID- 20013069 TI - Documentation of pain care processes does not accurately reflect pain management delivered in primary care. AB - BACKGROUND: Researchers and quality improvement advocates sometimes use review of chart-documented pain care processes to assess the quality of pain management. Studies have found that primary care providers frequently fail to document pain assessment and management. OBJECTIVES: To assess documentation of pain care processes in an academic primary care clinic and evaluate the validity of this documentation as a measure of pain care delivered. DESIGN: Prospective observational study. PARTICIPANTS: 237 adult patients at a university-affiliated internal medicine clinic who reported any pain in the last week. MEASURES: Immediately after a visit, we asked patients to report the pain treatment they received. Patients completed the Brief Pain Inventory (BPI) to assess pain severity at baseline and 1 month later. We extracted documentation of pain care processes from the medical record and used kappa statistics to assess agreement between documentation and patient report of pain treatment. Using multivariable linear regression, we modeled whether documented or patient-reported pain care predicted change in pain at 1 month. RESULTS: Participants' mean age was 53.7 years, 66% were female, and 74% had chronic pain. Physicians documented pain assessment for 83% of visits. Patients reported receiving pain treatment more often (67%) than was documented by physicians (54%). Agreement between documentation and patient report was moderate for receiving a new pain medication (k = 0.50) and slight for receiving pain management advice (k = 0.13). In multivariable models, documentation of new pain treatment was not associated with change in pain (p = 0.134). In contrast, patient-reported receipt of new pain treatment predicted pain improvement (p = 0.005). CONCLUSIONS: Chart documentation underestimated pain care delivered, compared with patient report. Documented pain care processes had no relationship with pain outcomes at 1 month, but patient report of receiving care predicted clinically significant improvement. Chart review measures may not accurately reflect the pain management patients receive in primary care. PMID- 20013070 TI - Comparing a self-administered measure of empathy with observed behavior among medical students. AB - PURPOSE: Studies show that measures of physician and medical students' empathy decline with clinical training. Presently, there are limited data relating self reported measures to observed behavior. This study explores a self-reported measure and observed empathy in medical students. METHOD: Students in the Class of 2009, at a university-based medical school, were surveyed at the end of their 2nd and 3rd year. Students completed the Jefferson Scale of Physician Empathy Student Version (JSPE-S), a self-administered scale, and were evaluated for demonstrated empathic behavior during Objective Structured Clinical Examinations (OSCEs). RESULTS: 97.6% and 98.1% of eligible students participated in their 2nd and 3rd year, respectively. The overall correlation between the JSPE-S and OSCE empathy scores was 0.22, p < 0.0001. Students had higher self-reported JSPE-S scores in their 2nd year compared to their 3rd year (118.63 vs. 116.08, p < 0.0001), but had lower observed empathy scores (3.96 vs. 4.15, p < 0.0001). CONCLUSIONS: Empathy measured by a self-administered scale decreased, whereas observed empathy increased among medical students with more medical training. PMID- 20013071 TI - Manometric changes of the lower esophageal sphincter after sleeve gastrectomy in obese patients. AB - INTRODUCTION: Laparoscopic sleeve gastrectomy has been accepted as an option for surgical treatment of obesity. After surgery, some patients present reflux symptoms associated with endoscopic esophagitis, therefore PPI's treatment must be indicated. PURPOSE: This study aims to evaluate the manometric characteristic of the lower esophageal sphincter (LES) before and after sleeve gastrectomy MATERIAL AND METHOD: This prospective study includes 20 patients submitted to esophageal manometry in order to determine the resting pressure, and total and abdominal LES length before and after the sleeve gastrectomy. Statistical variations on the LESP were validated according to Student's "t" test. RESULTS: Seventeen female and three male patients were included, with a mean age of 37.6 +/- 12.6 years. All patients reduced their body weight, from an initial BMI of 38.3 kg/m(2) to 28.2 kg/m(2) 6 months after surgery. No postoperative complications were observed in these patients. Preoperative mean LESP was 14.2 +/ 5.8 mmHg. Postoperative manometry decreased in 17/20 (85%), with a mean value of 11.2 +/- 5.7 mmHg (p = 0.01). Seven of them presented LESP <12 mmHg and ten patients <6 mmHg after the operation. Furthermore, the abdominal length and total length of the high pressure zone at the esophagogastric junction were affected. CONCLUSION: A sleeve gastrectomy produces an important decrease in LES pressure, which can in turn cause the appearance of reflux symptoms and esophagitis after the operation due to a partial resection of the sling fibers during the gastrectomy. PMID- 20013072 TI - On some aspects of the thermodynamic of membrane recycling mediated by fluid phase endocytosis: evaluation of published data and perspectives. AB - The theoretical and experimental description of fluid phase endocytosis (FPE) requires an asymmetry in phospholipid number between the two leaflets of the cell membrane, which provides the biomechanical torque needed to generate membrane budding. Although the motor force behind FPE is defined, its kinetic has yet to be determined. Based on a body of evidences suggesting that the mean surface tension is unlikely to be involved in endocytosis we decided to determine whether the cytosolic hydrostatic pressure could be involved, by considering a constant energy exchanged between the cytosol and the cell membrane. The theory is compared to existing experimental data obtained from FPE kinetic studies in living cells where altered phospholipid asymmetry or changes in the extracellular osmotic pressure have been investigated. The model demonstrates that FPE is dependent on the influx and efflux of vesicular volumes (i.e. vesicular volumes recycling) rather than the membrane tension of cells. We conclude that: (i) a relationship exists between membrane lipid number asymmetry and resting cytosolic pressure and (ii) the validity of Laplace's law is limited to cells incubated in a definite hypotonic regime. Finally, we discuss how the model could help clarifying elusive observations obtained from different fields and including: (a) the non-canonical shuttling of aquaporin in cells, (b) the relationship between high blood pressure and inflammation and (c) the mechanosensitivity of the sodium/proton exchanger. PMID- 20013073 TI - Ethanol production from the organic fraction obtained after thermal pretreatment of municipal solid waste. AB - In this work, the use of organic fraction from municipal solid waste (MSW) as substrate for ethanol production based on enzymatic hydrolysis was evaluated. MSW was subjected to a thermal pretreatment (active hygienization) at 160 degrees C from 5 to 50 min. The organic fiber obtained after 30 min was used as substrate in a simultaneous saccharification and fermentation (SSF) and fed-batch SSF process using cellulases and amylases. In a fed-batch mode with 25% (w/w) substrate loading, final ethanol concentration of 30 g/L was achieved (60% of theoretical). In these conditions, more than 160 L of ethanol per ton of dry matter could be produced from the organic fraction of MSW. PMID- 20013074 TI - Nitrogen source optimization for cellulase production by Penicillium funiculosum, using a sequential experimental design methodology and the desirability function. AB - The present study aimed at maximizing cellulase production by Penicillium funiculosum using sequential experimental design methodology for optimizing the concentrations of nitrogen sources. Three sequential experimental designs were performed. The first and the second series of experiments consisted of a 2(4) and a 2(3) factorial designs, respectively, and in the third one, a central composite rotational design was used for better visualizing the optimum conditions. The following nitrogen sources were evaluated: urea, ammonium sulfate, peptone, and yeast extract. Peptone and ammonium sulfate were removed from the medium optimization since they did not present significant statistical effect on cellulase production. The optimal concentrations of urea and yeast extract predicted by the model were 0.97 and 0.36 g/L, respectively, which were validated experimentally. By the use of the desirability function, it was possible to maximize the three main enzyme activities simultaneously, which resulted in values for FPase of 227 U/L, for CMCase of 6,917 U/L, and for beta-glucosidase of 1,375 U/L. These values corresponded to increases of 3.3-, 3.2-, and 6.7-folds, respectively, when compared to those obtained in the first experimental design. The results showed that the use of sequential experimental designs associated to the use of the desirability function can be used satisfactorily to maximize cellulase production by P. funiculosum. PMID- 20013075 TI - Comparative analysis of antigen-targeting sequences used in DNA vaccines. AB - Plasmid vectors can be optimized by including specific signals that promote antigen targeting to the major antigen presentation and processing pathways, increasing the immunogenicity and potency of DNA vaccines. A pVAX1-based backbone was used to encode the Green Fluorescence Protein (GFP) reporter gene fused either to ISG (Invariant Surface Glycoprotein) or to TSA (trans-sialidase) Trypanosoma brucei genes. The plasmids were further engineered to carry antigen targeting sequences, which promote protein transport to the extracellular space (secretion signal), lysosomes (LAMP-1) and to the endoplasmic reticulum (adenovirus e1a). Transfection efficiency was not affected by differences in the size between each construct as no differences in the plasmid copy number per cell were found. This finding also suggests that the addition of both ISG gene and targeting sequences did not add sensitive regions prone to nuclease attack to the plasmid. Cells transfected with pVAX1GFP had a significant higher number of transcripts. This could be a result of lower mRNA stability and/or a lower transcription rate associated with the bigger transcripts. On the other hand, no differences were found between transcript levels of each ISG-GFP plasmids. Therefore, the addition of these targeting sequences does not affect the maturation/stability of the transcripts. Microscopy analysis showed differences in protein localization and fluorescent levels of cells transfected with pVAX1GFP and ISG constructs. Moreover, cells transfected with the lamp and secretory sequences presented a distinct distribution pattern when compared with ISG protein. Protein expression was quantified by flow cytometry. Higher cell fluorescence was observed in cells expressing the cytoplasmic fusion protein (ISG GFP or TSA-GFP) compared with cells where the protein was transported to the lysosomal pathway. Protein transport to the endoplasmic reticulum does not lead to a decrease in the mean fluorescence values. The secretion signal was only effective when used in conjunction with TSA gene. Therefore, the characteristics of each protein (e.g., presence of transmembrane domains) might influence the efficacy of its cellular transport. This analysis constitutes a useful tool for the optimization of the design of DNA vaccines. PMID- 20013076 TI - Effect of ROCK inhibitor Y-27632 on normal and variant human embryonic stem cells (hESCs) in vitro: its benefits in hESC expansion. AB - The Rho associated coiled coil protein kinase (ROCK) dependent signaling pathway plays an important role in numerous physiological functions such as cell proliferation, adhesion, migration and inflammation. Human embryonic stem cells (hESCs) undergo differentiation and poor survival after single cell dissociation in culture thus limiting their expansion for cell based therapies. We evaluated the role of the selective ROCK inhibitor Y-27632 on hESC colonies and disassociated single hESCs from two different hESC lines. Karyotypically normal hESCs (HES3) and variant hESCs (BG01V) were treated with Y-27632 at 5, 10 and 20 muM concentrations for 72 h and its effects on hESC self renewal, colony morphology, cell cycle and pluripotency were evaluated. Increased cell proliferation of both HES3 and BG01V were observed for all three concentrations compared to untreated controls following passaging of cell clusters or dissociated single cells and some of these increases were statistically significant. Cell cycle assay demonstrated normal cell cycle progression with no peaks evident of apoptosis. No morphological differentiation was evident following treatment with the highest concentration of Y-27632 (20 muM) and the stemness related genes continued to be highly expressed in both HES3 and BG01V cells compared to untreated controls. The results confirmed that Y-27632 is a useful agent that aids in the expansion of undifferentiated hESC numbers for downstream applications in regenerative medicine. PMID- 20013078 TI - Gene expression-based diagnosis of efficacy of chemotherapy for breast cancer. AB - Development of a clear index to select drugs, i.e., accurate prediction of drug sensitivity, is important not only to obtain the maximum therapeutic effects of drugs, but also realize personalized medicine (tailor-made medicine). With the recent advancement in genome science represented by microarrays, molecular-level elucidation of many diseases including cancers has been progressing. It has been clarified that molecular information, such as gene expression profiles of cancer cells and gene polymorphisms in individual patients, affects not only cancer development and progression, but also therapeutic and adverse effects. The establishment of a therapeutic method by clinical application of this information has been progressing, in which the therapeutic effects of drugs are accurately predicted, and the maximum effects are obtained corresponding to cancer properties and patients' characteristics. PMID- 20013077 TI - A mimic of phosphorylated prolactin inhibits human breast cancer cell proliferation via upregulation of p21 waf1. AB - A mimic of phosphorylated prolactin (S179D PRL) inhibits mouse normal mammary HC11 cell proliferation through the upregulation of the vitamin D receptor (VDR) and p21. Here, we investigated whether S179D PRL also inhibited growth of human breast cancer MCF7 cells via VDR and p21. Western blots showed that S179D PRL upregulated VDR and p21 after the cells were incubated with S179D PRL for 3 days. These effects were blocked by the MAP kinase blocker PD98059 (25 MUM), indicating that MAPK plays a role in VDR and p21 upregulation. To confirm whether VDR contributes to p21 upregulation, we used two constructs that express luciferase. One (p21 VDRE Luc) has the vitamin D response element (VDRE) in the p21 promoter region; the other (p21 NO-VDRE Luc) does not. The results show that S179D PRL upregulated p21 VDRE Luc activity in p21 VDRE Luc-transfected cells more than in p21 NO-VDRE-transfected cells, indicating that S179D PRL upregulated p21 via VDR. A cell proliferation assay showed that S179D PRL inhibits cell proliferation in a dose-dependent manner. PMID- 20013079 TI - Effect of auxiliary substances on complexation efficiency and intrinsic dissolution rate of gemfibrozil-beta-CD complexes. AB - The studies reported in this work are aimed to elucidate the ternary inclusion complex formation of gemfibrozil (GFZ), a poorly water-soluble drug, with beta cyclodextrin (beta-CD) with the aid of auxiliary substances like different grades of povidone(s) (viz. PVP K-29/32, PVP K-40, Plasdone S-630, and Polyplasdone XL), organic base (viz. triethanolamine), and metal ion (viz. MgCl(2).6H(2)O), by investigating their interactions in solution and solid state. Phase solubility studies were carried out to evaluate the solubilizing power of beta-cyclodextrin, in association with various auxiliary substances, to determine the apparent stability constant (K (C)) and complexation efficiency (CE) of complexes. Improvement in K (C) values for ternary complexes clearly proves the benefit of the addition of auxiliary substances to promote CE. Of all the approaches used, the use of polymer Plasdone S-630 was found to be the most promising approach in terms of optimum CE and K (C). GFZ-beta-CD (1:1) binary and ternary systems were prepared by kneading and lyophilization methods. The ternary systems clearly signified superiority over binary systems in terms of CE, solubility, K (C), and reduction in the formulation bulk. Optimized ternary system of GFZ-beta-CD Plasdone S-630 prepared by using lyophilization method indicated a significant improvement in intrinsic dissolution rate when compared with ternary kneaded system. Differential scanning calorimetry, X-ray diffraction, Fourier transform infrared, scanning electron microscopy, and proton nuclear magnetic resonance were carried out to characterize the binary and optimized ternary complex. The results suggested the formation of new solid phases, eliciting strong evidences of ternary inclusion complex formation between GFZ, beta-CD, and Plasdone S-630, particularly for lyophilized products. PMID- 20013080 TI - Direct compression behavior of low- and high-methoxylated pectins. AB - The objective of this study was to evaluate possible usefulness of pectins for direct compression of tablets. The deformation behavior of pectin grades of different degree of methoxylation (DM), namely, 5%, 10%, 25%, 35%, 40%, 50%, and 60% were, examined in terms of yield pressures (YP) derived from Heckel profiles for both compression and decompression and measurements of elastic recovery after ejection. All pectin grades showed a high degree of elastic recovery. DM 60% exhibited most plastic deformation (YP 70.4 MPa) whereas DM 5% (104.6 MPa) and DM 10% (114.7 MPa) least. However, DM 60% gave no coherent tablets, whereas tablet tensile strengths for DM 5% and DM 10% were comparable to Starch 1500. Also, Heckel profiles were similar to Starch 1500. For sieved fractions (180-250 and 90 125 mum) of DM 25% and DM 40% originating from the very same batch, YPs were alike, indicating minor effects of particle size. These facts indicate that DM is important for the compaction behavior, and batch-to-batch variability should also be considered. Therefore, pectins of low degree of methoxylation may have a potential as direct compression excipients. PMID- 20013081 TI - Solid self-microemulsifying formulation for candesartan cilexetil. AB - Sparingly, water-soluble drugs such as candesartan cilexetil offer challenges in developing a drug product with adequate bioavailability. The objective of the present study was to develop and characterize self-microemulsifying drug delivery system (SMEDDS) of candesartan cilexetil for filling into hard gelatin capsules. Solubility of candesartan cilexetil was evaluated in various nonaqueous careers that included oils, surfactants, and cosurfactants. Pseudoternary phase diagrams were constructed to identify the self-microemulsification region. Four self microemulsifying formulations were prepared using mixtures of oils, surfactants, and cosurfactants in various proportions. The self-microemulsification properties, droplet size, and zeta potential of these formulations were studied upon dilution with water. The optimized liquid SMEDDS formulation was converted into free flowing powder by adsorbing onto a solid carrier for encapsulation. The dissolution characteristics of solid intermediates of SMEDDS filled into hard gelatin capsules was investigated and compared with liquid formulation and commercial formulation to ascertain the impact on self-emulsifying properties following conversion. The results indicated that solid intermediates showed comparable rate and extent of drug dissolution in a discriminating dissolution medium as liquid SMEDDS indicating that the self-emulsifying properties of SMEDDS were unaffected following conversion. Also, the rate and extent of drug dissolution for solid intermediates was significantly higher than commercial tablet formulation. The results from this study demonstrate the potential use of SMEDDS as a means of improving solubility, dissolution, and concomitantly the bioavailability. PMID- 20013082 TI - Prevalence and isotypic complexity of the anti-Chinese hamster ovary host cell protein antibodies in normal human serum. AB - Host cell-derived protein impurities may be present at low levels in biopharmaceutical products. Antibodies to host cell proteins are present in individuals with no known exposure to these products. In this study, antibodies to drug process-specific Chinese hamster ovary host cell-derived proteins (CHO HCP) were measured in unexposed individuals using a validated enzyme-linked immunosorbent assay. Samples that tested positive for anti-CHO-HCP reactivity were further characterized to determine the isotypes and IgG subclasses expressed in each positive individual. The specificity of the detected anti-CHO-HCP antibody isotypes was confirmed by the competitive inhibition assay and the uncoated plate specificity testing. These antibody characterization experiments revealed that the prevalent anti-CHO-HCP antibody subclasses were predominantly IgG1 (present in 66.6% of individuals) and IgG2 (60%), with 33.3% positive for IgG3 while IgG4 was detected in low abundance. Forty percent (40%) of the individuals with IgG type reactivity were also identified as IgM-positive. Anti CHO-HCP-specific IgE was not detected in the assays used in this study. Some individuals exhibited single isotype anti-CHO-HCP reactivity; others contained a combination of multiple antibody isotypes. Data presented in this study demonstrate the isotypic complexity and the high prevalence of anti-CHO-HCP antibodies in human serum with no known exposure to CHO cell-derived therapeutic biologics. PMID- 20013084 TI - IRE1alpha controls cyclin A1 expression and promotes cell proliferation through XBP-1. AB - IRE1 is a conserved dual endoribonuclease/protein kinase that is indispensable for directing the endoplasmic reticulum (ER) stress response in yeast, flies, and worms. In mammalian systems, however, the precise biological activities carried out by IRE1alpha are unclear. Here, molecular and chemical genetic approaches were used to control IRE1 activity in a number of prostate cancer cell lines and the resulting impact on gene transcription, cell survival, and proliferation was examined. Modulating IRE1alpha activity had no transcriptional effect on the induction of genes classically associated with the ER stress response (Grp78 and CHOP) or cell survival when confronted with ER stress agents. Rather, IRE1alpha activity was positively correlated to proliferation. Since Xbp-1 mRNA is the sole known substrate for IRE1 endoribonuclease activity, the role of this transcription factor in mediating proliferation was examined. Repressing total Xbp-1 levels by siRNA techniques effectively slowed proliferation. In an effort to identify IRE1/XBP-1 targets responsible for the cell cycle response, genome wide differential mRNA expression analysis was performed. Consistent with its ability to sense ER stress, IRE1alpha induction led to an enrichment of ER-Golgi, plasma membrane, and secretory gene products. An increase in cyclin A1 expression was the only differentially expressed cell cycle regulatory gene found. Greater cyclin A protein levels were consistently observed in cells with active IRE1alpha and were dependent on XBP-1. We conclude that IRE1alpha activity controls a subset of the ER stress response and mediates proliferation through tight control of Xbp-1 splicing. PMID- 20013083 TI - Molecular targets of dietary phenethyl isothiocyanate and sulforaphane for cancer chemoprevention. AB - Development of cancer is a long-term and multistep process which comprises initiation, progression, and promotion stages of carcinogenesis. Conceivably, it can be targeted and interrupted along these different stages. In this context, many naturally occurring dietary compounds from our daily consumption of fruits and vegetables have been shown to possess cancer preventive effects. Phenethyl isothiocyanate (PEITC) and sulforaphane (SFN) are two of the most widely investigated isothiocyanates from the crucifers. Both have been found to be very potent chemopreventive agents in numerous animal carcinogenesis models as well as cell culture models. They exert their chemopreventive effects through regulation of diverse molecular mechanisms. In this review, we will discuss the molecular targets of PEITC and SFN potentially involved in cancer chemoprevention. These include the regulation of drug-metabolizing enzymes phase I cytochrome P450s and phase II metabolizing enzymes. In addition, the signaling pathways including Nrf2 Keap 1, anti-inflammatory NFkappaB, apoptosis, and cell cycle arrest as well as some receptors will also be discussed. Furthermore, we will also discuss the similarities and their potential differences in the regulation of these molecular targets by PEITC and SFN. PMID- 20013085 TI - Processing of limb kinematics in the interpositus nucleus. AB - Neural representations of limb movement kinematic parameters are common among central nervous system structures involved in motor control, such as the interpositus nucleus of the cerebellum. Much experimental evidence indicates that neurons in the interpositus may encode limb kinematic parameters both during active, voluntary actions and during limb motion imposed passively, which entrains only sensory afferents. With respect to the sensory processing of information related to movement kinematics, we show that interpositus neuronal activity can parse out the directional from the scalar component (i.e., the movement speed) of the velocity vector. Moreover, a differential role for the anterior and posterior portion of interpositus in encoding these parameters emerged from these data, since the activity of the posterior interpositus was specifically associated to changes of movement speed. Limb movement representations in the interpositus nucleus may be instrumental for the control of goal-directed movements such as shaping hand during grasping or precise foot placement during gait. Finally, we discuss the idea that sensory information about the movement kinematics contribute to both feedback and anticipatory processes for limb movement control. PMID- 20013086 TI - Combined cetuximab and reirradiation for locoregional recurrent and inoperable squamous cell carcinoma of the head and neck. AB - PURPOSE: To investigate the feasibility, toxicity, and efficacy of external-beam reirradiation (Re-RT) combined with cetuximab for patients with inoperable and recurrent squamous cell carcinoma of the head and neck (SCCHN). PATIENTS AND METHODS: Seven patients with inoperable recurrence of SCCHN after adjuvant or definitive radiotherapy (RT) and simultaneous or sequential cisplatin-based chemotherapy for primary SCCHN were treated between August and December 2008 with Re-RT (1.8 Gy/fraction to 50.4 Gy) and cetuximab (400 mg/m(2) initial dose in the 1st week, and then 250 mg/m(2) once weekly). Recurrence had to be located at least > or = 50% in the preirradiated field. Long term toxicity from previous treatment was recorded before Re-RT as a baseline value. Acute and late toxicity derived from the experimental regimen were recorded every week during RT, and then every 3 months. Efficacy was assessed with repeated imaging using response evaluation criteria in solid tumors (RECIST) and clinical examinations 8-12 weeks after end of the treatment and every 3 months thereafter (Tables 1 and 2). RESULTS: Only mild localized mucositis occurred in all patients. Two patients developed a grade 3 acneiform rash related to cetuximab. After treatment one patient developed a grade 2 trismus, another showed grade 3 abacterial salivary gland inflammation with severe pain requiring opioid medication. Two patients achieved a complete response after 7 months, one remained stable, three progressed, and one died from pneumonia without having restaging magnetic resonance imaging. CONCLUSION: A second course of RT combined with cetuximab in patients with inoperable, recurrent HNSCC proved to be feasible with mild or moderate toxicity and encouraging response to treatment. PMID- 20013087 TI - Proton-beam therapy for hepatocellular carcinoma associated with portal vein tumor thrombosis. AB - BACKGROUND AND PURPOSE: The prognosis of patients with advanced hepatocellular carcinoma (HCC) with portal vein tumor thrombosis (PVTT) is extremely poor, as effective treatment options are limited. The authors performed a retrospective review to evaluate the efficacy of proton-beam therapy (PBT) for patients presenting with PVTT in the setting of HCC. PATIENTS AND METHODS: Between February 1991 and September 2005, 35 patients with HCC and tumor thrombi in the main trunk or major branches of the portal vein presented for consideration of PBT. Their tumor sizes ranged from 25 mm to 130 mm (median, 60 mm). A median total dose of 72.6 GyE in 22 fractions was delivered over 31 days to a target volume that encompassed both the primary hepatic lesion and the PVTT. RESULTS: 32 patients were progression-free during a median follow-up period of 21 months (range, 2-88 months) and three patients experienced disease progression. Local progression-free survival rates were 46% at 2 years and 20% at 5 years, and the median local progression-free survival was 21 months. Acute toxicity > or = grade 3 was observed in three patients, and no patient experienced late toxicity > or = grade 3. None of the patients had to discontinue treatment as a result of toxicity. CONCLUSION: PBT improved local control and significantly prolonged survival in HCC patients with PVTT. PMID- 20013088 TI - Perioperative intensity-modulated brachytherapy for refractory orbital rhabdomyosarcomas in children. AB - PURPOSE: To evaluate the feasibility and toxicity of perioperative intensity modulated brachytherapy (IMBT) as well as functional outcome in children with therapy-refractory orbital rhabdomyosarcomas (RMS). PATIENTS AND METHODS: Since 1993, children with therapy-refractory orbital RMS have been treated by a multidisciplinary approach combining function-preserving, mostly R1 tumor resection and perioperative IMBT at the University Hospital of Schleswig Holstein, Germany. All children with orbital RMS, who were enrolled in this multidisciplinary treatment protocol between 1993 and 2002, were prospectively assessed with respect to evaluation of side effects and functional outcome. RESULTS: Ten children (six boys, four girls) were included. Median age was 6.5 years (range, 1-19 years) at the beginning of our treatment and 6.0 years (range 1-17 years) at diagnosis. All children were in Intergroup Rhabdomyosarcoma Study Group (IRSG) group III and had embryonal subtype. Estimated 5-year survival was 62% +/- 18%. There was no radiation-related toxicity grade 3 or 4 observed. The eyes were primarily preserved in all cases. One child underwent secondary orbital exenteration 10 months after completion of IMBT. Visual acuity could be preserved apart from one child developing significant visual deterioration due to radiation cataract grade 2. The cosmetic results were good or very good in eight and moderate in two children. Four children died of their disease. CONCLUSION: This interdisciplinary, individually tailored and function-preserving treatment procedure has proven to be a well-tolerated therapeutic option in cases with refractory orbital RMS. It provides both improvement of local tumor control and quality of life. PMID- 20013089 TI - Intensity-modulated arc therapy with simultaneous integrated boost in the treatment of primary irresectable cervical cancer. Treatment planning, quality control, and clinical implementation. AB - PURPOSE: To report on the planning procedure, quality control, and clinical implementation of intensity-modulated arc therapy (IMAT) delivering a simultaneous integrated boost (SIB) in patients with primary irresectable cervix carcinoma. PATIENTS AND METHODS: Six patients underwent PET-CT (positron emission tomography-computed tomography) and MRI (magnetic resonance imaging) before treatment planning. Prescription (25 fractions) was (1) a median dose (D(50)) of 62, 58 and 56 Gy to the primary tumor (GTV_cervix), primary clinical target volume (CTV_cervix) and its planning target volume (PTV_cervix), respectively; (2) a D(50) of 60 Gy to the PET-positive lymph nodes (GTV_nodes); (3) a minimal dose (D(98)) of 45 Gy to the planning target volume of the elective lymph nodes (PTV_nodes). IMAT plans were generated using an anatomy-based exclusion tool with the aid of weight and leaf position optimization. The dosimetric delivery of IMAT was validated preclinically using radiochromic film dosimetry. RESULTS: Five to nine arcs were needed to create valid IMAT plans. Dose constraints on D(50) were not met in two patients (both GTV_cervix: 1 Gy and 3 Gy less). D(98) for PTV_nodes was not met in three patients (1 Gy each). Film dosimetry showed excellent gamma evaluation. There were no treatment interruptions. CONCLUSION: IMAT allows delivering an SIB to the macroscopic tumor without compromising the dose to the elective lymph nodes or the organs at risk. The clinical implementation is feasible. PMID- 20013090 TI - Prospective study of exclusive strontium-/yttrium-90 beta-irradiation of primary and recurrent pterygia with no prior surgical excision. Clinical outcome of long term follow-up. AB - PURPOSE: To evaluate the consecutive treatment results regarding pterygium recurrence and the efficacy of exclusive strontium-/yttrium-90 beta-irradiation for primary and recurrent pterygia and to analyze the functional outcome. PATIENTS AND METHODS: Between October 1974 and December 2005, 58 primary and 21 recurrent pterygia were exclusively treated with strontium-/yttrium-90 beta irradiation with doses ranging from 3,600 to 5,500 cGy. The follow-up time was 46.6 +/- 26.7 months, with a median of 46.5 months. RESULTS: The treatment led to a size reduction in all pterygia (p < 0.0001). Neither recurrences nor side effects were observed during therapy and follow-up in this study. Best-corrected visual acuity increased (p = 0.0064). Corneal astigmatism was reduced in recurrent pterygia (p = 0.009). CONCLUSION: Exclusive strontium-/yttrium-90 beta irradiation of pterygia is a very efficient and well-tolerated treatment, with remarkable aesthetic and rehabilitative results in comparison to conventional treatments, especially for recurrent lesions which have undergone prior surgical excision. PMID- 20013091 TI - Radiation- and chemoinduced multidrug resistance in colon carcinoma cells. AB - BACKGROUND AND PURPOSE: Radiation can induce multidrug resistance (MDR) and thus interfere with simultaneous or subsequent chemotherapy. In SW620 colon carcinoma cells, the interrelation of various biological endpoints of MDR was analyzed and the potential of fractionated irradiation and chemoselection to evoke MDR was compared. MATERIAL AND METHODS: To induce/select an MDR phenotype, SW620 were exposed to either 27 Gy in 1.8-Gy daily fractions or to 50% inhibiting concentrations of doxorubicin or cisplatin, given over 6-15 weeks. Expression of genes involved in MDR, including glutathione metabolism, was determined by semiquantitative RT-PCR (reverse transcription-polymerase chain reaction). Efflux was analyzed by flow cytometry after staining with rhodamine-123 or 5 chloromethyl fluorescein diacetate. Apoptosis was monitored after pulse exposure to doxorubicin or cisplatin. Colony-forming assays were performed under continuous drug exposure. RESULTS: A pronounced gene induction was found in MRP2 after cisplatin selection and up to 3 weeks after radiation. LRP was activated only shortly after radiation. Radiation enhanced rhodamine-123 efflux to a similar extent as short-term chemoselection but not as much as long-term drug exposure. Drug-induced apoptosis was slightly delayed in preirradiated cells. Clonogenic growth in the progeny of irradiated cells was less sensitive to cisplatin but not to doxorubicin. CONCLUSION: Fractionated radiation can induce an MDR phenotype in SW620. However, long-term drug exposure establishes a more efficient selection. Various endpoints are not fully concordant regarding the extent of MDR. Posttranscriptional modifications, pleiotropic regulation, and alternative pathways may cause these discrepancies. PMID- 20013092 TI - Overestimation of low-dose radiation in intensity-modulated radiotherapy with sliding-window technique. AB - PURPOSE: To analyze different control-system limitations on the measured dose distributions in low-dose regions of simplified intensity fields with an electronic portal imaging device to ascertain the optimal settings for the control-system limitations in the planning system. MATERIAL AND METHODS: The authors created one field with an "optimal fluence" of intensity 1.0 (full dose) and one field with intensity 0.0 (no dose) in the central part of the field. The influence of different dose rates (DRs) and maximum leaf speeds (LS) on the calculated and measured dose and dose profiles were analyzed. RESULTS: Good agreement between calculated and measured dose in the case of a field of intensity 1.0 was found. For the field with intensity 0.0, the measured dose was 20-60% lower than the dose calculated by the "actual fluence". The results were found dependent on the DR and LS. CONCLUSION: The overestimation in regions of optimal intensity 0.0 by the planning system cannot be resolved by the user. Taking the measured dose in the region of desired intensity 1.0 and other technical limitations (like beam hold interrupts or spikes in the cross and longitudinal profiles) into consideration, the application of an LS of 2.5 cm/s and a DR of 500 MU/min is recommended in order to minimize radiation dose applied to organs at risk, which are located in regions of low intensity, like, for example, the spinal cord. PMID- 20013093 TI - First results of the federal quality assurance group ("Arztliche Stelle") in radiotherapy in Baden-Wurttemberg: part 2. AB - PURPOSE: Part 2 of this study presents the results of the clinical audits of the "Arztliche Stelle" (AS) concerning process quality in the radiation therapy units in Baden-Wurttemberg, Germany. MATERIAL AND METHODS: The process quality of nine main test groups (laws, organizations, technical equipment, GMP [Good Medical Practice] principles, radiotherapy, radiation procedure, follow-up, discharge report, physics) were questioned by the AS commission and evaluated in a four stage classification (1-4; none to grave deficiencies). RESULTS: In 18 institutes (81%), the medical and physical-technical inspection, including radiation protection for patients and staff, revealed very high to high quality, safe treatment commensurate with current standards was guaranteed once the suggestions for improvement given by the experts had been implemented. In four institutes (18%), the inspection showed a still satisfactory quality but a shorter period for reinspection was recommended. The most frequent comments/deficiencies were found in staff structures (32%), in continuous education/training for staff (32%), in informed patient consent (41%), follow-up (45%), and/or a unclearly formulated discharge report for the referring physicians (45%). CONCLUSION: The medical and physical-technical examination of radiation treatment in the vast majority (81%) of the radiation therapy institutes in Baden-Wurttemberg showed a very high or high quality. Most of the comments and deficiencies concerned the sector of systematic continued training for staff, giving clear explanations to patients, structured follow-up, and the state of the medical discharge reports. These deficiencies, however, do not represent any immediate danger to the patient through any deficiencies in therapy. The experience gained by the AS could act as a model for quality improvement and could also be transmitted to other medical sectors to retain quality standards. The various specialist associations are called upon to gradually adopt the suggestions for quality improvements which are based on concrete as-is analyses by steadily aligning practice with theory. This would enable the steadily rising demands of quality assurance to be sustainably implemented through practicable models. PMID- 20013094 TI - Secular trend and risk factors for antimicrobial resistance in Escherichia coli isolates in Switzerland 1997-2007. AB - BACKGROUND: Antibacterial resistance in Escherichia coli isolates of urinary infections, mainly to fluoroquinolones, is emerging. The aim of our study was to identify the secular trend of resistant E. coli isolates and to characterize the population at risk for colonization or infections with these organisms. PATIENTS AND METHODS: Retrospective analysis of 3,430 E.coli first isolates of urine specimens from patients admitted to the University Hospital Basel in 1997, 2000, 2003, and 2007. RESULTS: Resistance to ciprofloxacin, trimethoprim/sulfamethoxazole, and amoxicillin/clavulanate has increased over the 10-year study period (from 1.8% to 15.9%, 17.4% to 21.3%, and 9.5% to 14.5%, respectively). A detailed analysis of the 2007 data revealed that independent risk factors for ciprofloxacin resistance were age (5.3% < 35 years of age to 21.9% in patients > 75 years; odds ratio [OR] 1.29 per 10 years, 95% confidence interval [CI] 1.15-1.45, p < 0.001) and male gender (OR 1.59, 95% CI 1.05-2.41, p = 0.04). In contrast, nosocomial E. coli isolates were associated with lower odds of ciprofloxacin resistance (OR 0.51, 95% CI 0.28-0.67, p < 0.001). The frequency of resistant isolate rates was not influenced by the clinical significance (i.e., colonization vs urinary tract infection, UTI) or by whether the urine was taken from a urinary catheter. Importantly, the increase in ciprofloxacin resistance paralleled the increase in ciprofloxacin consumption in Switzerland (Pearson's correlation test R(2)= 0.998, p = 0.002). Of note, resistance was less frequent in isolates sent in by general practitioners. However, after adjustment for age and gender, only resistance against amoxicillin/clavulanate was found to be less frequent (OR 0.34, 95% CI 0.16-0.92, p = 0.03). CONCLUSION: Our study reveals that resistance rates have been increasing during the last decade. Published resistance rates may lack information due to important differences regarding age, gender, and probable origin of the isolates. Empirical therapy for UTI should be guided more on individual risk profile and local resistance data than on resistance data banks. PMID- 20013095 TI - Septicemia progressing to fatal hepatic dysfunction in an cirrhotic patient after oral ingestion of Photobacterium damsela: a case report. PMID- 20013098 TI - Magnetic resonance-thoracic ductography: imaging aid for thoracic surgery and thoracic duct depiction based on embryological considerations. AB - PURPOSE: We describe the optimal protocol of magnetic resonance-thoracic ductography (MRTD) and provide examples of thoracic ducts (TD) and various anomalies. The anatomical pathway of the TD was analyzed based on embryological considerations. METHODS: A total of 78 subjects, consisting of noncancer adults and patients with esophageal cancer and lung cancer, were enrolled. The MRTD protocol included a long echo time and was based on emphasizing signals from the liquid fraction and suppressing other signals, based on the principle that lymph flow through the TD appears hyperintense on T2-weighted images. The TD configuration was classified into nine types based on location [right and/or left side(s) of the descending aorta] and outflow [right and/or left venous angle(s)]. RESULTS: MRTD was conducted in 78 patients, and the three-dimensional reconstruction was considered to provide excellent view of the TD in 69 patients, segmentalization of TD in 4, and a poor view of the TD in 5. MRTD achieved a visualization rate of 94%. Most of the patients had a right-side TD that flowed into the left venous angle. Major configuration variations were noted in 14% of cases. Minor anomalies, such as divergence and meandering, were frequently seen. CONCLUSION: MRTD allows noninvasive evaluation of TD and can be used to identify TD configuration. Thus, this technique is considered to contribute positively to safer performance of thoracic surgery. PMID- 20013097 TI - Preservation solution for lung transplantation. AB - Despite the increasing success of lung transplantation as the mainstay therapeutic modality for end-stage lung disease, the 1-year survival rate after lung transplantation remains 80% and primary graft dysfunction (PGD) accounts for 30% of mortality. Ischemia-reperfusion injury has been identified as one of the main causes of PGD, and thus significant efforts have been made to optimize the methods for lung preservation in an attempt to minimize lung injury during the period of ischemia. The composition of the lung preservation solution used in the pulmonary artery flush has been considered to be the key to successful lung preservation, and many lung transplant programs have been shifting the use of the preservation solution from the intracellular fluid type to the extracellular fluid type because of preferable posttransplant lung function with the latter. This review summarizes the experimental and clinical studies on lung preservation, particularly focusing attention on the preservation solution being employed for clinical lung transplantation. PMID- 20013099 TI - Symptomatic gastroesophageal reflux disease after lung transplantation. AB - PURPOSE: Gastroesophageal reflux disease (GERD) is associated with allograft dysfunction after lung transplantation (LTX). We attempted to identify outcomes in LTX recipients with clinical evidence of GERD. METHODS: Retrospective review of 162 LTX recipients at our institution between January 1994 and June 2006 was performed. GERD was confirmed in symptomatic patients by esophagogastroduodenoscopy (EGD) and/or esophagography. Occurrence of biopsy proven obliterative bronchiolitis (OB) and bronchiolitis obliterans syndrome (BOS) were analyzed. Kaplan-Meier analysis of survival and Cox proportional hazard analysis of risk factors were performed. RESULTS: GERD was diagnosed in 21 (13%) of patients, usually following LTX (71%). There was no difference in mean survival (1603 +/- 300 vs. 1422 +/- 131 days; log rank P > 0.05), or development of OB (5% vs. 6%, respectively; P > 0.05) in patients with GERD compared with patients without GERD. However, there was correlation between GERD and BOS (P = 0.01). CONCLUSIONS: Symptomatic GERD is increased following LTX. Patients with symptomatic GERD demonstrated an increased incidence of BOS, but survival was not affected in this study. More sensitive and specific diagnostic tools should be implemented in all LTX recipients to investigate the impact of symptomatic and silent GERD and thus improve outcomes after LTX. PMID- 20013100 TI - Acute pulmonary embolism after cerebral infarction associated with a mobile thrombus in the ascending aorta. AB - Although the causes of stroke are diverse, thromboembolism due to a mobile aortic thrombus is rare. We describe a surgical case of acute massive pulmonary embolism after critical cerebral infarction associated with a mobile ascending aortic thrombus in a 52-year-old woman. Concomitant surgical removal of the aortic thrombus and pulmonary embolectomy was performed successfully, and the patient has been stable without recurrent thromboembolic complications after 18 months of follow-up. PMID- 20013101 TI - Radio-guided thoracoscopic surgery with (99m)Tc-methoxy-isobutylisonitrile for treating an ectopic mediastinal parathyroid adenoma in an adolescent girl. AB - Here we report the case of a 17-year-old girl treated for an ectopic mediastinal parathyroid adenoma using (99m)Tc-methoxy-isobutyl-isonitrile (MIBI) and radio guided thoracoscopic surgery. Her serum calcium and intact parathyroid hormone levels were elevated, and an anterior mediastinal tumor was noted on her chest computed tomography scan. At 2 h before surgery, the patient was given an injection of 370 MBq of (99m)Tc-MIBI. Because it was not possible visually to distinguish the tumor from the mature thymic tissue in which it was embedded, a search for the tumor was carried out using a gamma probe under thoracoscopy. Thereafter, the left thymus, including the radioactivity-positive tumor, was resected. We suggest that radio-guided thoracoscopic surgery with (99m)Tc-MIBI is an excellent procedure for treating ectopic mediastinal parathyroid adenomas embedded in the mature thymus of adolescents and young adults. This approach could be especially useful for young female patients, given its cosmetic advantage. PMID- 20013102 TI - Transsternal transpericardial closure of a postpneumonectomy bronchial fistula in a patient who underwent pneumonectomy because of a war injury. AB - Most common causes of intrathoracic empyema include pulmonary infections and postoperative bronchopleural fistulas complicating a lung surgical resection, mainly pneumonectomy, as a result of the failure of the bronchial stump to heal. A 22-year-old Serbian patient presented with chronic posttraumatic empyema. Two years before during a war, he experienced chest injury due to a firearm wound, with massive intrathoracic bleeding and need for emergency left pneumonectomy. Empyema with a bronchopleural fistula occurred during the postoperative course. The patient underwent left open window thoracostomy with a daily bandage change. Here we report the treatment of the bronchopleural fistula using sequential surgical approach including transsternal transpericardial closure of the fistula followed by reconstruction of the chest wall with a regional muscle flap. Our case report highlights the feasibility and efficacy of the transsternal surgical approach to treat postpneumonectomy bronchopleural fistula, thereby avoiding the direct approach to the bronchial stump through the infected pneumonectomy cavity. PMID- 20013103 TI - End-stage localized bronchiectasis mimicking a hemangioma. AB - We report a case of localized bronchiectasis mimicking a hemangioma demonstrated on enhanced chest computed tomography. Left lower lobectomy following selective bronchial arterial embolization was successful. Pathology evaluation showed markedly dilated bronchial arteries along a dilated bronchus displacing the lateral and posterior basal segments. PMID- 20013104 TI - Bilateral multiple sclerosing hemangiomas of the lung. AB - This report presents a case of bilateral multiple sclerosing hemangiomas of the lung in a 73-year-old woman. A computed tomography (CT) scan of the chest showed a total of three well-defined small nodules in the right and left lower lobes of the lung. Because malignant metastatic lung tumors were highly suspected, a wedge resection of the left lower lobe was performed to obtain a definitive diagnosis. Histopathologically, two tumors of the left lower lobe were composed of epithelial-like cuboidal cells covering the surface of papillary protrusions and sheets of round to polygonal cells underneath the epithelial-like cells. The final diagnosis was that both tumors were sclerosing hemangiomas. After surgery the residual lesion of the right lower lobe was carefully followed by chest CT. The size of the right lung nodule did not change over the course of 9 years, and no new lesion has emerged. PMID- 20013105 TI - Primary malignant melanoma of the lung with rapid progression. AB - This report presents a case of primary malignant melanoma of the lung with rapid progression in a 68-year-old man. During a regular checkup, a chest roentgenogram revealed an abnormal shadow in the left lower lung field. Computed tomography (CT) of the chest revealed an ill-defined tumor shadow measuring 4 x 3 cm in the left upper lobe. A CT-guided lung biopsy suggested a large-cell carcinoma, and left upper lobectomy was performed. Histopathologically, the tumor was comprised of malignant epithelial tumor cells with large amounts of acidophilic cytoplasm and prominent nuclei. A dark brown pigment was observed in the tumor cells. Immunohistochemical staining was positive for S-100 protein and HMB45; staining for cytokeratin, CAM5.2, and chromogranin was negative. The final diagnosis was malignant melanoma of the lung. Two months later, multiple brain metastases developed, and he died of the disease 6 months after the surgery. PMID- 20013106 TI - Video-thoracoscopic management of a postpneumonectomy bronchopleural fistula. AB - Bronchopleural fistula (BPF) after pneumonectomy is a well-known, dreaded complication associated with a high mortality rate. We present a case of video assisted thoracoscopic management of a silent, small BPF. After thoracoscopic removal of all debris and inflammatory tissues, the identified BPF was successfully sealed by thoracoscopic instillation of tissue glue. This approach may be an effective alternative and promising option for treating small bronchopleural fistulas accompanied by empyema after pneumonectomy. PMID- 20013107 TI - Foldback technique for aortic anastomosis of free right internal thoracic artery in coronary artery bypass grafting. AB - We herein report a new technique that we term "foldback plasty" for stenosis-free proximal anastomosis of free arterial grafts without using any other interposition materials. The arterial graft is first anastomosed to the aorta in side-to-side fashion, leaving 1 cm of remnant tissue at the proximal end. The reverse side of the graft is then opened longitudinally, starting at the proximal end, to just distal to the aortic anastomosis site. The new proximal graft flap is folded back and sutured onto the graft to close the longitudinal opening. The proximal anastomosis site is enlarged only by the graft tissue, and stenosis can be avoided even if the graft has a small caliber. PMID- 20013108 TI - [Therapy standards after flexor tendon and nerve injuries of the hand: results from a survey of German centres for hand surgery]. AB - BACKGROUND: There is little evidence for the ideal aftercare of combined nerve and flexor tendon injuries of the hand. The aim of this study was to elicit whether concomitant nerve injuries are changing the individual treatment plans after flexor tendon repair in a survey of German centres for hand surgery. METHODS: A questionnaire about aftercare of isolated and combined nerve and flexor tendon injuries of the hand was distributed to members of three German Societies of hand, trauma and plastic surgery. RESULTS: Isolated flexor tendon injuries in zones II to IV are treated by early mobilization in all centres, whereas isolated digital nerve repair is usually followed by immobilization (10% no immobilization, 22.5% up to 1 week, 52.5% for 2 weeks and 15% for 3 weeks). The duration of immobilization increases with lesions of the median or ulnar nerves by about 1 week. In 55% of cases concomitant nerve injury does not influence the early onset of dynamic splinting and mobilization after flexor tendon injuries. CONCLUSION: There seem to be no uniform treatment guidelines for flexor tendon repair if concomitant nerve injury is present. Against the background of the current literature early controlled mobilization after tendon and nerve repair seems to be justified. PMID- 20013109 TI - Insulin requirements throughout pregnancy in women with type 1 diabetes mellitus: three changes of direction. AB - AIMS/HYPOTHESIS: The aim of the study was to analyse the insulin requirements of women with type 1 diabetes mellitus throughout pregnancy. METHODS: We have examined the weekly mean blood glucose (mmol/l), insulin requirements (U kg(-1) day(-1)) and total insulin requirements (U/day) in 65 women with type 1 diabetes mellitus and tight metabolic control since before pregnancy (HbA(1c) < or =6.0%). RESULTS: Both insulin requirement and total insulin requirement displayed a peak in week 9, a nadir in week 16 and a second peak in week 37. For the change in insulin requirement (4.08% per week) and in total insulin requirement (5.19% per week), the sharpest slope was observed from week 16 to week 37. However, two changes of direction took place in the first 11 weeks and eight out of nine episodes of severe hypoglycaemia requiring treatment with glucagon or i.v. glucose took place in the first 16 weeks. CONCLUSIONS/INTERPRETATION: Pregnant women with type 1 diabetes mellitus and tight metabolic control since before pregnancy displayed changes in insulin requirement and total insulin requirement with successive changes of direction. The sharpest slope was observed between 16 and 37 weeks, but insulin requirements were more unstable in the first 16 weeks. This information could help patients and physicians to react to changes in glycaemic pattern in a prompt and adequate way. PMID- 20013110 TI - Validation of the Patient Global Impression of Improvement (PGI-I) for urogenital prolapse. AB - INTRODUCTION AND HYPOTHESIS: Currently, there is no global outcome assessment index in prolapse research. Patient Global Impression of Improvement (PGI-I) has only been validated in incontinence. Our aim was to validate its use following prolapse surgery. METHODS: Women with prolapse were recruited from waiting lists and assessed objectively (pelvic organ prolapse quantification system (POP-Q)). Quality of life (QoL) was assessed with prolapse QoL questionnaire (pQoL). Patient goal achievement (visual analogue scale (VAS)) determined subjective satisfaction and PGI-I indicated overall satisfaction. We established construct validity of PGI-I by correlating final PGI-I response with other measures of response, measured at 1 year: (POP-Q/pQoL/VAS) RESULTS: There was excellent test retest reliability and correlation between PGI-I and other outcome measures. CONCLUSION: We believe this is the first study validating PGI-I as a global index of response to prolapse surgery. This may be a valuable addition not only in clinical practice but also in trials comparing surgical interventions. PMID- 20013111 TI - A new model of the disrupted latent inhibition in C57BL/6J mice after bupropion treatment. AB - RATIONALE: Schizophrenia is characterized by disturbances in attention and information processing that can be measured by latent inhibition (LI). Research has implicated significant aberrations in dopaminergic (DA) neurotransmission in this disorder. OBJECTIVES: The objectives of this study were as follows: to probe whether bupropion disrupts LI; to compare its efficacy to the effects of GBR12783 (specific DA uptake inhibitor) and to amphetamine (DA releaser); to test if antipsychotics would reverse LI deficits induced by bupropion, GBR12783, and amphetamine; and to probe if rolipram (phosphodiesterase-4 inhibitor), which increases cyclic AMP (cAMP) similarly to antipsychotics, effectively corrects drug-induced LI deficits. Based on its efficacy in drug addiction, we also asked if bupropion could block the effect of amphetamine. METHODS: LI was measured in a conditioned emotional response procedure by comparing suppression of drinking in response to a noise in C57BL/6J mice. Mice previously received 0 (nonpreexposed) or 40 noise exposures (preexposed) followed by two or four noise-foot shock pairings. RESULTS: Bupropion abolished LI in mice, which was corrected by rolipram, but not by haloperidol and clozapine. GBR12783 and amphetamine, but not antidepressants, also disrupted LI, and this was reversed by antipsychotics and rolipram. Both bupropion and amphetamine disrupted LI via conditioning session. Paradoxically, bupropion and GBR12783 also blocked the amphetamine-induced LI deficit. CONCLUSIONS: Efficacy of rolipram but not antipsychotics to reverse the effects of bupropion suggests novel cAMP-dependent and D(2) receptor-independent mechanisms of the bupropion-induced LI deficit. Further detailed biochemical analysis of bupropion-induced LI deficit might be a fruitful approach in developing new antipsychotics. PMID- 20013112 TI - The effects of infusions of CART 55-102 into the basolateral amygdala on amphetamine-induced conditioned place preference in rats. AB - RATIONALE: The affective aspects of D: -amphetamine (AMPH) may be mediated, in part, by cocaine- and amphetamine-regulated transcript (CART) peptides in the basolateral amygdala (BLA). The formation of context-drug associations produces either conditioned place preference (CPP) or conditioned place aversion (CPA). OBJECTIVES: The aim of the present study was to determine whether intra-BLA infusions of CART 55-102 are either rewarding or aversive and modulate AMPH reward. MATERIALS AND METHODS: Rats were implanted with bilateral cannulae in the BLA, were subjected to place conditioning, and were tested for CPP or CPA. Rats were conditioned with either intra-BLA infusions of artificial cerebral spinal fluid or one of three dose of CART 55-102 (1, 2, or 4 microg/side), intra-BLA infusions of a subrewarding dose of CART 55-102 (1 microg/side) plus injections of a subrewarding dose of AMPH (0.1 mg/kg, i.p.), or intra-BLA infusions of an aversive dose of CART 55-102 (4 microg/side) plus injections of a rewarding dose of AMPH (1.0 mg/kg, i.p.). RESULTS: Intra-BLA infusions of 2 microg/side CART 55 102 produced CPP, 4 microg/side produced CPA, and 1 microg/side produced neither CPP nor CPA. Intra-BLA infusions of a subrewarding dose of CART 55-102 (1 microg/side) plus injections of a subrewarding dose of AMPH (0.1 mg/kg, i.p.) produced CPP. Intra-BLA infusions of an aversive dose of CART 55-102 (4 microg/side) plus injections of a rewarding dose of AMPH (1.0 mg/kg, i.p.) produced neither CPP nor CPA. CONCLUSIONS: Both the affective properties of intra BLA CART 55-102 and its ability to either facilitate or block AMPH reward are dose dependent. PMID- 20013113 TI - Coadministration of intravenous nicotine and oral alcohol in rats. AB - RATIONALE AND OBJECTIVES: Alcohol and nicotine are the most commonly abused drugs, and they are often taken together. We have developed a procedure in which rats self-administer nicotine intravenously and alcohol orally during the same operant session. METHODS: Male Wistar rats were trained to self-administer alcohol (12%, w/v; 0.19 ml/alcohol delivery) or implanted with jugular catheters and trained to self-administer nicotine (30 microg/kg i.v./infusion) by pressing a lever or were trained to self-administer both drugs, some with alcohol first, and others with nicotine first. The effects of extinction of responding for either or both drugs in animals trained to coadminister alcohol and nicotine and the effects of alcohol and nicotine primes on reinstatement were also determined. RESULTS: Animals readily coadministered alcohol and nicotine concurrently. Access to alcohol reduced nicotine self-administration significantly. When responding for alcohol was extinguished with nicotine still available, extinction of alcohol seeking was slowed significantly. In rats trained to coadminister nicotine and alcohol, priming with nicotine or alcohol reinstated extinguished responding for both drugs. Reinstatement of extinguished nicotine or alcohol seeking by, respectively, nicotine or alcohol priming was unaffected by continued access to the other drug. CONCLUSIONS: These results show that rats will self-administer relevant amounts of intravenous nicotine and oral alcohol concurrently. They also provide further support for the important relationship between nicotine and alcohol. PMID- 20013115 TI - Melatonin treatment for eastward and westward travel preparation. AB - INTRODUCTION: Melatonin is recommended for hastening adaptation to phase shift, but there is little information on appropriate formulations. MATERIALS AND METHODS: We evaluated the efficacy of three melatonin formulations for circadian phase advance and delay: (a) 3 mg regular release (RR), (b) 3 mg sustained release (SR), and (c) 3 mg surge-sustained release (SSR; consisting of 1 mg RR and 2 mg SR). Circadian phase was assessed by salivary melatonin dim light melatonin onset (DLMO) or offset (MelOff) using thresholds of (1) 1.0 pg/ml and (2) mean baseline + 2 standard deviations (BL + 2SD). Subjects spent from Tuesday evenings until Thursday in the laboratory. Melatonin (or placebo) was administered at 1600 hours (phase advance) Wednesday, with DLMO assessment on Tuesday and Thursday and at 0600 hours (phase delay) Wednesday, with DLMO assessment Tuesday, Wednesday, and MelOff Thursday morning. Phase advances using the 1.0 pg/ml DLMO were as follows: placebo, 0.73 h; RR, 1.23 h (p < 0.003); SR, 1.44 h (p < 0.0002); SSR, 1.16 h (p < 0.012), with no difference between formulations. RESULTS AND DISCUSSION: Similar but smaller phase advances were found with BL + 2SD. Using MelOff, posttreatment phase position for the RR formulation was delayed compared to placebo by 1.12 h (p < 0.012), 1.0 pg/ml, and 0.75 h (p < 0.036), BL+2SD. Phase shifts for the SR and SSR conditions could not be determined due to persistent high melatonin levels during sampling times. Similar phase advances were induced by all formulations, and slow clearance of slow release preparations impeded the determination of phase delays. CONCLUSION: Appropriately timed 0.5 mg melatonin doses may avoid these problems. PMID- 20013114 TI - Modulation of prepulse inhibition through both M(1) and M (4) muscarinic receptors in mice. AB - RATIONALE: Muscarinic cholinergic M(1) and M(4) receptors may participate in schizophrenia's etiology and have been proposed as targets for antipsychotic medications. OBJECTIVE: Here, we investigated the involvement of these receptors in behavioral measures pertinent to schizophrenia using knockout mice lacking M(1) receptors (M(1)-/-), M(4) receptors (M(4)-/-), or both (M(1)-/-M(4)-/-). METHODS: We measured prepulse inhibition (PPI) of startle without drugs and after treatment with scopolamine (0.32-1.8 mg/kg), xanomeline (3.2 mg/kg), oxotremorine (0.032-0.1 mg/kg), clozapine (1.0-5.6 mg/kg), or haloperidol (0.32-3.2 mg/kg). RESULTS: In female (but not male) mice, combined deletion of both M(1) and M(4) receptors decreased PPI relative to wild-type mice, while knockout of either receptor alone had no significant effect. Scopolamine disrupted PPI in wild-type and M(4)-/- mice, but not in female M(1)-/-M(4)-/- or female M(1)-/- mice. When administered before scopolamine, xanomeline restored PPI in wild-type mice and M(1)-/- mice, but not in M(4)-/- mice. In contrast, pretreatment with oxotremorine increased PPI regardless of genotype. Effects of clozapine and haloperidol on PPI were not hindered by either mutation. CONCLUSIONS: Deletion of both M(1) and M(4) receptors can disrupt PPI, suggesting that (at least partially redundant) M(1) and M(4) receptor-dependent functions are involved in sensorimotor gating mechanisms. PPI-disrupting effects of muscarinic antagonists appeared dependent upon M(1) receptor blockade. Our data also suggest that xanomeline exerts antipsychotic-like effects mainly through M(4) receptor stimulation, while stimulation of non-M(1)/M(4) subtypes may also have antipsychotic potential. Finally, our results do not support a role of M(1)/M(4) receptors in mediating antipsychotic-like effects of clozapine. PMID- 20013117 TI - Extracorporeal shock-wave lithotripsy and garlic consumption: a lesson to learn. AB - The first case of a kidney haematoma after extracorporeal shock-wave lithotripsy (SWL) in a patient with nephrolithiasis who was taking aged garlic extraction is reported. Patient was treated conservatively without the need of any intervention. Urologists should be aware that herbal products including garlic, ginkgo, and ginseng have been associated with potential increased bleeding. The present case emphasises the need to specifically seek out a history of herbal use in presurgical patients. It is suggested that herbal medications should be discontinued up to 15 days prior to urologic surgery or SWL to minimise the risk of complications. PMID- 20013116 TI - Qualitative differences between C57BL/6J and DBA/2J mice in morphine potentiation of brain stimulation reward and intravenous self-administration. AB - RATIONALE: The C57BL/6J (C57) and DBA/2J (DBA) mice are the most common genotypes used to identify chromosomal regions and neurochemical mechanisms of interest in opioid addiction. Unfortunately, outside of the oral two-bottle choice procedure, limited and sometimes controversial evidence is available for determining their relative sensitivity to the rewarding effects of morphine. OBJECTIVES: The purpose of this study was to utilize classically accepted models of drug abuse liability to determine relative susceptibility to the rewarding effects of morphine. METHODS: The ability of morphine or amphetamine to potentiate lateral hypothalamic brain stimulation and intravenous morphine self-administration (across three doses in a fixed ratio schedule and at the highest dose in progressive ratio schedules) was investigated in both genotypes. RESULTS: In both measures, C57 and DBA mice differed dramatically in their response to morphine. Morphine potentiated rewarding stimulation in the C57 mice but antagonized it in the DBA mice. Consistent with these findings, intravenous morphine did not serve as a positive reinforcer in DBA mice under conditions that were effective in the C57 mice using a fixed ratio schedule and failed to sustain levels of responding sufficient to maintain a constant rate of drug intake under a progressive ratio schedule. In contrast, amphetamine potentiated the rewarding effects of brain stimulation similarly in the two genotypes. CONCLUSIONS: These findings provide strong evidence that morphine is rewarding in the C57 genotype and not in the DBA genotype. Understanding their relative susceptibility is important given the prominence of these genotypes in candidate gene identification and gene mapping. PMID- 20013118 TI - Antibacterial and biofilm removal activity of a podoviridae Staphylococcus aureus bacteriophage SAP-2 and a derived recombinant cell-wall-degrading enzyme. AB - Antibacterial and biofilm removal activity of a new podoviridae Staphylococcus aureus bacteriophage (SAP-2), which belongs to the phi29-like phage genus of the Podoviridae family, and a cell-wall-degrading enzyme (SAL-2), which is derived from bacteriophage SAP-2, have been characterized. The cell-wall-degrading enzyme SAL-2 was expressed in Escherichia coli in a soluble form using a low-temperature culture. The cell-wall-degrading enzyme SAL-2 had specific lytic activity against S. aureus, including methicillin-resistant strains, and showed a minimum inhibitory concentration of about 1 microg/ml. In addition, this enzyme showed a broader spectrum of activity within the Staphylococcus genus compared with bacteriophage SAP-2 in its ability to remove the S. aureus biofilms. Thus, the cell-wall-degrading enzyme SAL-2 can be used to prevent and treat biofilm associated S. aureus infections either on its own or in combination with other cell-wall-degrading enzymes with anti-S. aureus activity. PMID- 20013119 TI - Scaling up microbial fuel cells and other bioelectrochemical systems. AB - Scientific research has advanced on different microbial fuel cell (MFC) technologies in the laboratory at an amazing pace, with power densities having reached over 1 kW/m(3) (reactor volume) and to 6.9 W/m(2) (anode area) under optimal conditions. The main challenge is to bring these technologies out of the laboratory and engineer practical systems for bioenergy production at larger scales. Recent advances in new types of electrodes, a better understanding of the impact of membranes and separators on performance of these systems, and results from several new pilot-scale tests are all good indicators that commercialization of the technology could be possible within a few years. Some of the newest advances and future challenges are reviewed here with respect to practical applications of these MFCs for renewable energy production and other applications. PMID- 20013120 TI - Synthesis of gamma-aminobutyric acid (GABA) by Lactobacillus plantarum DSM19463: functional grape must beverage and dermatological applications. AB - Agriculture surplus were used as substrates to synthesize gamma-aminobutyric acid (GABA) by Lactobacillus plantarum DSM19463 for the manufacture of a functional beverage or as a novel application for dermatological purposes. Dilution of the grape must to 1 or 4% (w/v) of total carbohydrates favored higher cell yield and synthesis of GABA with respect to whey milk. Optimal conditions for synthesizing GABA in grape must were: initial pH 6.0, initial cell density of Log 7.0 cfu/mL, and addition of 18.4 mM L-glutamate. L. plantarum DSM19463 synthesized 4.83 mM of GABA during fermentation at 30 degrees C for 72 h. The fermented grape must also contain various levels of niacin, free minerals, and polyphenols, and Log 10.0 cfu/g of viable cells of L. plantarum DSM19463. Freeze dried preparation of grape must was applied to the SkinEthic(R) Reconstructed Human Epidermis or multi-layer human skin model (FT-skin tissue). The effect on transcriptional regulation of human beta-defensin-2 (HBD-2), hyaluronan synthase (HAS1), filaggrin (FGR), and involucrin genes was assayed through RT-PCR. Compared to GABA used as pure chemical compound, the up-regulation HBD-2 was similar while the effect on the expression of HAS1 and FGR genes was higher. PMID- 20013121 TI - Monoclonal gammopathy of undetermined significance related pyoderma gangrenosum successfully treated with autologous peripheral blood stem cell transplantation. PMID- 20013122 TI - [Grayzone lymphoma. Clinical relevance]. AB - Malignant lymphomas are classified into different entities according to their morphology, immunohistochemical parameters and clinical behavior. Several important pathogenetic events can be assigned to certain lymphoma entity types. Nevertheless, some cases present overlapping morphologic and immunohistochemical characteristics and a clear-cut diagnosis cannot be made. This is particularly the case with aggressive lymphomas for which a clear distinction cannot be made between the entities of diffuse large cell lymphoma/Burkitt lymphoma or primary mediastinal B cell lymphoma/classic Hodgkin's lymphoma. In order to redress this situation, two new gray zone entities were introduced in the WHO 2008 classification. Until further knowledge regarding the therapy, behavior and prognosis of these gray zone lymphomas has been gained, they should continue to be considered as distinct entities. PMID- 20013125 TI - Staphylococcus warneri ventriculoperitoneal shunt infection: failure of diagnosis by ventricular CSF sampling. AB - BACKGROUND: The definite diagnosis of hydrocephalus valve infection is generally made by cerebrospinal fluid (CSF) sampling via the valve reservoir, which is considered to be more dependable than that of the CSF obtained by lumbar puncture. CASE REPORT: We treated a 17-year-old boy with an intra-abdominal pseudocyst due to ventriculoperitoneal shunt infection caused by Staphylococcus warneri whose ventricular CSF, obtained via the valve reservoir, was repeatedly sterile thus causing a considerable delay in the management of the complication. DISCUSSION AND CONCLUSIONS: S. warneri constitutes an emergent contaminant of catheters and prostheses. We found only a detailed report of S. warneri infection of a ventriculoatrial shunt. If manifestations of peritoneal involvement in shunted patients would occur, the attention should be shifted to the distal component of the shunt hardware, even in the presence of a normal ventricular CSF as happened in our case to avoid unnecessary delay in diagnosis and management. PMID- 20013124 TI - Surgery for extratemporal nonlesional epilepsy in children: a meta-analysis. AB - PURPOSE: Previous small studies have demonstrated that seizure outcomes following surgery for extratemporal lobe epilepsy (ETLE) in children are worse than those for temporal lobe epilepsy. We have conducted a meta-analysis of the available literature to better understand ETLE surgical outcomes in children. METHODS: We searched PubMed (1990-2009) for appropriate studies using the following terms: ETLE, ETLE surgery, ETLE surgery outcome, frontal lobe epilepsy, occipital lobe epilepsy, and parietal lobe epilepsy. Our collected data included patient age at seizure onset and surgery, the cerebral lobe involved with epileptogenesis, MRI findings, predominant seizure semiology, intracranial monitoring use (electrode implantation), epileptic region histopathology, and postoperative seizure outcome. Statistical analysis was performed to determine associations among these variables and postoperative outcome. RESULTS: Ninety-five patients from 17 studies satisfied the inclusion criteria. Pathological findings (p = 0.039) and seizure type (p = 0.025) were significantly associated with outcome: A larger proportion of patients with cortical dysplasia and complex partial seizures experienced better outcomes. Age at surgery (p = 0.073) and the cerebral resection site (p = 0.059) were marginally associated with seizure outcome. CONCLUSIONS: This study confirms previous reports: Surgical outcomes for ETLE epilepsy are significantly worse than those for temporal lobe epilepsy. The reasons for this difference may include the diffuse nature of the pathology involved in ETLE, difficulty in localizing the seizure focus in young children, and involvement of "eloquent" nonresectable cortex in epileptogenesis. Because of the reporting variability among different epilepsy centers, more uniform protocols are necessary for fair evaluation and comparison of outcomes among the different centers. PMID- 20013126 TI - Association between food and nutrient intakes and cognitive capacity in a group of institutionalized elderly people. AB - BACKGROUND: Some authors have indicated that the cognitive decline may be due to an inadequate nutritional status. AIM OF STUDY: To determine the association between food and nutrient intakes and cognitive capacity score in a group of institutionalized elderly people. METHODS: The study subjects were 178 elderly (> or = 65 years of age) institutionalized people from the Madrid region. The diets of these subjects were recorded using the precise weighing method over a 7-day period, and their cognitive capacity assessed using the Short Portable Mental Status Questionnaire (SPMSQ). Subjects were grouped into those who did not incur errors (SPMSQ = 0) and who incurred one or more errors (SPMSQ > 0). Since an association was seen between the SPMSQ test score and age (r = 0.2030; p < 0.01), the subjects were also grouped according to whether they were above or below the percentile 50 (P(50)) for this variable (83 years). RESULTS: The subjects with no errors in the SPMSQ test (32%) consumed greater quantities of cereals, eggs, oils, and fats. After adjusting for energy intake and educational level an inverse association was seen between fish and vegetable consumption and cognitive capacity score. In addition, these subjects had greater intakes of carbohydrates, polyunsaturated fatty acids, riboflavin, and vitamins C, D, and E. After adjusting for energy intake, a negative relationship was found between cognitive capacity score and the intake of fibre, vitamin B(6), and folic acid. CONCLUSION: In general, the subjects of our study showed an adequate mental capacity, but those who made no errors in the SPMSQ test had more satisfactory diets. This shows the importance of the diet in the maintenance of cognitive function. PMID- 20013127 TI - Forensic age estimation in living subjects based on the ossification status of the medial clavicular epiphysis as revealed by thin-slice multidetector computed tomography. AB - Determination of the stage of ossification of the medial clavicular epiphysis is a crucial part of age estimation in criminal proceedings when evaluating individuals with completed hand ossification. In order to ensure a maximum of accuracy in forensic age estimation practise, it is recommended to perform thin slice CT scans; but to date there exist no reference data on the bone development of the region in question based on thin-slice computed tomography. In this retrospective study, the authors evaluated thin-slice multidetector CT images of 592 individuals aged between 10 and 35 years produced in the University Hospital of Munster. The ossification status of the medial epiphysis of the clavicle could be reliably determined in 502 cases using the classification of stages by Schmeling et al. In male individuals, stage 2 was first noted at age 14, in female individuals at age 13. Stage 3 was first achieved by male individuals at age 17, by female individuals at age 16. The occurrence of stage 4 was first found in both sexes at the age of 21. In either sex, the earliest observation of stage 5 was at age 26. The findings are basically in line with those from the only CT-based study on the subject in question using the same classification of five stages, except from the fact that in the present study, stage 5 first occurs at age 26, which is 4 or 5 years later than what was found in the CT study using 7 mm slices in the majority of cases. This vast difference may be explained through the partial volume effect occurring with thick-slice CT images by a visual deception of the epiphyseal scar occurring with stage 4. PMID- 20013128 TI - Postoperative laryngoscopy in thyroid surgery: proper timing to detect recurrent laryngeal nerve injury. AB - BACKGROUND: There is currently a lack of consensus to support the proper timing for postoperative laryngoscopy that is reliable to diagnose recurrent laryngeal nerve palsy (RLNP) after thyroid surgery. The purpose of this study was to investigate the impact of different time intervals of fiber-optic nasolaryngoscopy (FNL) on the diagnosis of RLNP. METHOD: FNL was performed postoperatively at day 0 (T1), at second day post-op (T2), and +2 weeks (T3). For patients with RLNP, repeated examinations were performed at +2 (T4), +6 (T5), and +12 months (T6). RESULTS: Four hundred thirty-four patients appear for postoperative FNL, providing 825 nerves at risk. Permanent RLNP occurred in 0.7%, temporary RLNP in 6.7%. RLNP rate was 6.4% at T1, 6.7% at T2, 4.8% at T3, 2.5% at T4, 0.8% at T5, and 0.7% at T6. Full recovery of vocal cord function was confirmed after rehabilitation in 87.5% of cases at T5 and 89% in T6. T2 was significantly superior to T3 in terms of diagnosis of RLNP (P < 0.05). Of patients at T2, 10.7% did not see any reason to FNL because of their normal voice register. CONCLUSION: FNL is essential for the detection of vocal cord paralysis after thyroidectomy. We report different time evaluation criteria of vocal cord motility with great and significant variability of results. Second day post-op inspection of the larynx (T2) is suggested. Symptomatic voice assessment is insufficient. PMID- 20013130 TI - Effect of sponge volume on the performance of down-flow hanging sponge system treating UASB reactor effluent. AB - The performance of down-flow hanging sponge (DHS) system treating UASB reactor effluent at different sponge volume of 38.2, 28.7, 19.1 and 9.6% was investigated. The hydraulic retention time (HRT) and flow rate were kept constant at 2.6 h and 0.46 m(3)/day, respectively. The results obtained indicated that reducing the sponge volume of the DHS system from 38.2 to 19.1% significantly decreased the removal efficiency of COD(total) from 80 +/- 8 to 62 +/- 14%; COD(soluble) from 71.2 +/- 10 to 53.7 +/- 18% and COD(particulate) from 86 +/- 10 to 62 +/- 15%. With further reduction of the sponge volume down to 9.6%, the removal efficiency was dropped to 54 +/- 15% for COD(total); 44.1 +/- 14% for COD(soluble) and 42 +/- 12% for COD(particulate). Likewise, the nitrification efficiency and fecal coliform (FC) removal was strongly affected by decreasing the sponge volume of the DHS system, i.e., the nitrification efficiency was dropped by a value of 48.3% when reducing the sponge volume of the DHS system from 38.2 to 19.1%. At sponge volume of 38.2%, the DHS system removes 2.6 +/- 0.3 log(10)/100 ml of FC which is significantly higher than that at sponge volume of 28.7, 19.1 and 9.6%. Accordingly, it is recommended to design and operate such a system at a bulk sponge volume not exceeding 38.2% of the total reactor volume and at an HRT of 2.6 h. PMID- 20013129 TI - Schimke immunoosseous dysplasia: defining skeletal features. AB - Schimke immunoosseous dysplasia (SIOD) is an autosomal recessive multisystem disorder characterized by prominent spondyloepiphyseal dysplasia, T cell deficiency, and focal segmental glomerulosclerosis. Biallelic mutations in swi/snf-related, matrix-associated, actin-dependent regulator of chromatin, subfamily a-like 1 (SMARCAL1) are the only identified cause of SIOD, but approximately half of patients referred for molecular studies do not have detectable mutations in SMARCAL1. We hypothesized that skeletal features distinguish between those with or without SMARCAL1 mutations. Therefore, we analyzed the skeletal radiographs of 22 patients with and 11 without detectable SMARCAL1 mutations. We found that patients with SMARCAL1 mutations have a spondyloepiphyseal dysplasia (SED) essentially limited to the spine, pelvis, capital femoral epiphyses, and possibly the sella turcica, whereas the hands and other long bones are basically normal. Additionally, we found that several of the adolescent and young adult patients developed osteoporosis and coxarthrosis. Of the 11 patients without detectable SMARCAL1 mutations, seven had a SED indistinguishable from patients with SMARCAL1 mutations. We conclude therefore that SED is a feature of patients with SMARCAL1 mutations and that skeletal features do not distinguish who of those with SED have SMARCAL1 mutations. PMID- 20013131 TI - Changes in management of acute pancreatitis before and after the publication of evidence-based practice guidelines in 2003. AB - BACKGROUND: The Japanese Guidelines for the Management of Acute Pancreatitis was published in 2003. However, the impact of the guidelines on physicians' practice patterns has not been well known. METHODS: To examine the current clinical practices in the management of acute pancreatitis, we conducted a questionnaire survey with members of three societies involved in the treatment of pancreatic diseases and abdominal emergency medical care. Questions included diagnostic and treatment processes considered important in the management of acute pancreatitis in addition to demographic data, experience in medical care, and areas of specialty of respondents. We also examined changes in the treatment of acute pancreatitis before and after publication of the Guidelines. RESULTS: Of 1,000 society members to whom questionnaires were mailed, 590 responded. Respondents who had read the Guidelines also handled significantly more cases in the most recent 3 years. A variety of changes were observed in the performance of clinical practices before and after publication of the Guidelines. The use of amylase in the assessment of severity decreased significantly, while its use for determination of severity scores increased significantly after publication of the Guidelines. In treatment, use of a nasogastric tube in mild and moderate cases deceased after the Guidelines. The frequency of prophylactic use of antibiotics decreased with mild pancreatitis after publication of the Guidelines. CONCLUSIONS: Although it is difficult to attribute these changes to the direct influence of the Guidelines, several changes were observed in performance of clinical practices in accordance with recommendations of the Guidelines. PMID- 20013132 TI - Implementation of real-time RT-PCR for detection of human metapneumovirus and its comparison with enzyme immunoassay. AB - Human metapneumovirus (hMPV) is responsible for outbreaks of bronchiolitis in winter and early spring in young children. Due to the relatively recent discovery of hMPV, the diagnostic opportunities are limited, while differential diagnosis with respiratory syncytial virus (RSV) remains important. We validated the RT-PCR by comparing various methods of RNA extraction, one-step RT-PCR kits and primer probe combinations. The optimized RT-PCR was evaluated using 47 nasopharyngeal aspirates (NPAs) collected from children younger than 5 years, with clinically suspected RSV infection. The evaluated RT-PCRs were also compared to a commercially available hMPV enzyme immunoassay (EIA). We found 8.5% hMPV positivity with both RT-PCRs, in agreement with published literature. hMPV EIA showed positive and indeterminate results in 17% and 8.5%, respectively, of the tested NPAs. Positive RT-PCR samples were positive or indeterminate by hMPV EIA. Samples that were positive for RSV and influenza A virus interfered with the hMPV EIA. In conclusion, although RT-PCR is already a valuable tool for diagnosing hMPV infections, further optimization of the RT-PCR method is recommended. The hMPV EIA kit shows poor specificity and therefore needs further improvement. PMID- 20013134 TI - Density functional studies of the substituent effect on absorption and emission properties of 1, 8-naphthalimide derivatives. AB - Using TD-PBE1PBE/6-31G* and TD-B3LYP/6-31G* approaches, we calculated the absorption and emission spectra of 1,8-naphthalmide derivatives in gas-phase. The geometric structures optimized by HF/6-31G* and B3LYP/6-31G* models and the absorption and emission maxima were in good agreement with existed experimental measurements. It was also found that the lowest singlet states corresponded mainly to the electronic transition from the highest occupied orbital (HOMO) to the lowest unoccupied molecular orbital (LUMO). Intramolecular charge transfer occurred between substituents and naphthalimic rings. Study also showed that most compounds with low absorption excitation energies had low vertical ionization potentials. Finally, the delocalization electronic energies between substituents and naphthalimic rings of isomers were investigated to obtain further sight into their stability. PMID- 20013135 TI - Mechanism of CDK5 activation revealed by steered molecular dynamics simulations and energy calculations. AB - In the current work, CDK5/p25 complexes were pulled apart by applying external forces with steered molecular dynamics (SMD) simulations. The crucial interactions between the kinase and the activation protein were investigated and the SMD simulations showed that several activation-relevant motifs of CDK5 leave p25 in sequence during the pulling and lead to an apo-CDK2 like CDK5 structure after separation. Based on systematic examination of hydrogen bond breaking and classical MD/molecular mechanics-generalized Born/surface area) (MM-GBSA) calculations, a CDK5 activation mechanism by p25 is suggested. This is the first step towards the systemic development of CDK inhibitors and the mechanism proposed could lead to a better understanding of the protein-protein recognition characteristics between the kinase and its activator. PMID- 20013136 TI - Molecular docking and 3D-QSAR studies on triazolinone and pyridazinone, non nucleoside inhibitor of HIV-1 reverse transcriptase. AB - Nonnucleoside reverse transcriptase inhibitors (NNRTIs) are allosteric inhibitors of the HIV-1 reverse transcriptase. Recently a series of Triazolinone and Pyridazinone were reported as potent inhibitors of HIV-1 wild type reverse transcriptase. In the present study, docking and 3D quantitative structure activity relationship (3D QSAR) studies involving comparative molecular field analysis (CoMFA) and comparative molecular similarity indices analysis (CoMSIA) were performed on 31 molecules. Ligands were built and minimized using Tripos force field and applying Gasteiger-Huckel charges. These ligands were docked into protein active site using GLIDE 4.0. The docked poses were analyzed; the best docked poses were selected and aligned. CoMFA and CoMSIA fields were calculated using SYBYL6.9. The molecules were divided into training set and test set, a PLS analysis was performed and QSAR models were generated. The model showed good statistical reliability which is evident from the r2 nv, q2 loo and r2 pred values. The CoMFA model provides the most significant correlation of steric and electrostatic fields with biological activities. The CoMSIA model provides a correlation of steric, electrostatic, acceptor and hydrophobic fields with biological activities. The information rendered by 3D QSAR model initiated us to optimize the lead and design new potential inhibitors. PMID- 20013137 TI - Histamine (re)uptake by astrocytes: an experimental and computational study. AB - Astrocytes participate in the clearance of neurotransmitters by their uptake and subsequent enzymatic degradation. Histamine as a polar and/or protonated molecule must use a carrier to be transported across the cell membrane, although a specific histamine transporter has not been elucidated, yet. In this work we upgraded the kinetic studies of histamine uptake into neonatal rat cultured type 1 astrocytes with quantum chemical calculations of histamine pKa values in conjunction with Langevin dipoles solvation model as the first step toward microscopic simulation of transport. Our results indicate that astrocytes transport histamine by at least two carrier mediated processes, a concentration gradient dependent passive and a sodium-dependent and ATP-driven active transport. We also demonstrated that histamine protonation states depend on the polarity of the environment. In conclusion we suggest that histamine, a polar molecule at physiological pH uses at least two different mechanisms for its uptake into astrocytes -an electrodiffusion and Na(+)-dependent and ouabain sensitive active process. We emphasize relevance of knowledge of histamines protonation states at the rate limiting step of its transport for microscopic simulation that will be possible when structure of histamine transporter is known. PMID- 20013138 TI - Treatment of resistant port wine stains (PWS) with pulsed dye laser and non contact vacuum: a pilot study. AB - The blanching of resistant port wine stains (PWS) with a pulsed dye laser (PDL) requires a large number of treatments, resulting in substantial discomfort to patients, many of them children. Pneumatic skin flattening (PSF - Serenity Pro) is a new technology that generates a vacuum over the skin and reduces pain in laser-based treatments of the skin, while creating contact between the skin and an upper window. The same technology can be utilized to increase skin blood fraction while operated in a non-contact mode. The objective of this study was to test the enhancement in the efficacy of PWS treatment with PDL and Serenity Pro while vacuum is being utilized in the non-contact, blood-enrichment mode. Fifteen patients with resistant PWS underwent 1-4 treatments (interval of 5-20 weeks) under general anesthesia with a 595-nm PDL at 10-14 J/cm(2), 1.5-3 ms pulse duration, and 7-mm spot size. Lesion blanching with DCD chilling and with vacuum were photographed and compared. Better blanching of various degrees was observed on resistant PWS with the blood-enrichment technique in seven out of 11 patients who returned for follow-up. There were no cases of decrease in efficacy. Blood enrichment with the Serenity Pro non-contact vacuum technology has the potential of enhancing the capability of treating resistant port wine stains in over 50% of cases. Further studies will better quantify the number of treatments necessary for better lesion clearance. The vacuum-assisted technique may be of particular importance in view of the fact that achieving complete lesion clearance remains a challenge in PWS treatments. PMID- 20013139 TI - Cisplatin-induced macroautophagy occurs prior to apoptosis in proximal tubules in vivo. AB - BACKGROUND: Autophagy is an intracellular bulk degradation process induced by cell starvation. Autophagy was recently reported to be induced by various stresses such as hypoxia, ischemia/reperfusion, toxins, and denatured proteins, and to affect cell survival and death. Light chain 3-II (LC3-II) is specifically located on double membrane-bound autophagosomes that envelop disused proteins or organelles. METHOD: Transgenic mice in which green fluorescent protein (GFP) was fused to LC3 (LC3-GFP) were administered cisplatin (20 mg/kg). After euthanasia at times between 0 and 72 h, kidneys were excised for immunohistochemical analyses. Microscopic examinations of the generated NRK-52E cell lines stably transfected with LC3-GFP, and Western blot analyses of NRK-52E cells, were undertaken after cisplatin treatment with or without autophagy inhibitors and beclin 1 small interfering RNA (siRNA). RESULTS: Autophagosomes increased in the proximal tubular cells of transgenic mice from 12 h after cisplatin injection (20 mg/kg). The time course for this was faster than those for tubular necrosis and apoptosis. Autophagosomes also increased in NRK-52E cells after cisplatin treatment, with the time course for this being faster than that for apoptosis. When autophagy was suppressed by autophagy inhibitors or beclin 1 siRNA, the level of apoptosis was also suppressed. CONCLUSION: Autophagy occurs in proximal tubular cells after cisplatin treatment and is involved in cell death in renal tubular injury. Our data suggest that autophagy is a kind of cell damage index and that cells with activated autophagy will be scavenged by apoptosis. PMID- 20013140 TI - Hypertensive encephalopathy in patients with chronic renal failure caused by stopping antihypertensive agents: a report of two cases. AB - We herein report two cases of patients with chronic kidney disease who developed hypertensive encephalopathy, which occurred after a sudden discontinuance of antihypertensive agents. Both patients underwent care at our hospital after experiencing neurological abnormalities. In both patients, magnetic resonance imaging (MRI) revealed edema in the cerebral white matter and cortices, basal ganglia, brainstem, and cerebellum. Though recently the number of reports about hypertensive encephalopathy has decreased, we describe two case reports and also review the pertinent literature. PMID- 20013143 TI - Weight-loss practices among university students in Mexico. AB - OBJECTIVE: To evaluate the prevalence of weight-loss practices among university students from Tlaxcala, Mexico. METHODS: A cross-sectional study of 2,651 university students was conducted. Logistic regression tests were used to estimate the probability of students trying to lose weight and successfully achieving weight loss. RESULTS: Nearly 40% of students attempted to lose weight, though only about 7% lost more than 10% of their body weight and maintained this weight loss during the time of the study. The methods used most were exercise and dieting, and those who dieted were more successful at losing weight. CONCLUSIONS: The high prevalence of weight-loss attempts and the poor outcomes with these weight-loss methods among this sample of university students is a public health concern. Universities should provide students with healthy weight-control approaches, which include offering information about healthier lifestyles, access to healthy food and opportunities to be physically active. PMID- 20013142 TI - The attitude toward tobacco dependence and barriers to discussing smoking cessation: a survey among Turkish general practitioners. AB - OBJECTIVES: To evaluate the general attitude of a sample of Turkish general practitioners (GPs) toward tobacco dependence and to assess their knowledge and behavior regarding smoking cessation (SC). METHODS: A self-administered questionnaire modified from WHO, Global Health Professional Survey was distributed to GPs, working in our district, Isparta. RESULTS: As much as 41% of GPs were current smokers. Ever smokers were generally less likely to agree with statements that would change their freedom to smoke in certain places. While 46% of GPs frequently inquired about tobacco use in their patients, 13.5% did not advise any of their patients to quit smoking during the month preceding the questionnaire. The most common barriers reported by GPs to discussing SC with their patients were as follows: considering the discussion not to be effective (57.8%), having low confidence in knowledge (48.1%), having unpleasant personal experience or considering it a thankless task (46.1%). CONCLUSIONS: It appears essential to reduce the number of GPs who smoke and to improve GP training on SC procedures for integrating SC treatment into primary care in Turkey. PMID- 20013145 TI - Meningeal carcinomatosis from penile squamous cell carcinoma. AB - We report herein a clinical case of a patient with meningeal carcinomatosis from penile squamous cell carcinoma. A 68-year-old man presented with mental changes, headaches, and unstable gait. Examinations revealed brain metastases and infiltration of the leptomeninges and subarachnoid space by carcinoma cells. Only 11 months earlier the patient had been diagnosed with penile squamous cell carcinoma of poor differentiation and had underwent subtotal penectomy and adjuvant chemotherapy and radiation. Infiltration of the central nervous system with penile cancer is extremely rare, and only five cases with brain metastases have been described to date. This is the first report of a patient with penile cancer spread to the leptomeninges. PMID- 20013144 TI - Sampling procedures and sample representativeness in a national telephone survey: a Portuguese example. AB - OBJECTIVES: This paper aims to assess the yielding of the sampling procedures and the representativeness of a sample evaluated in a national telephone survey in Portugal. METHODS: The Portuguese telephone book (2007/2008) was the sampling frame for subjects >39 years (n = 1,934). The time spent with interviews and unsuccessful contacts was quantified and the yielding of the contact attempts compared according different schedules. Demographic and health characteristics of the participants were compared with those of the Portuguese population. RESULTS: Among eligible individuals, the percentage of refusals was 40.4%, higher on Saturdays (45.2% vs. 30.7%) and evenings (45.2% vs. 36.7%). Evening interviews yielded a higher percentage of men (26.2% vs. 21.0%) and active workers (31.8% vs. 21.8%). Compared to the Portuguese population, our sample included a higher proportion of women (76.7% vs. 54.1%), elderly (>59 years: 61.4% vs. 46.2%) and more educated participants (>4 years: 38.3% vs. 26.3%). Sex- and age-specific estimates of smoking and obesity were similar to those in the Portuguese population. CONCLUSION: Despite the difficulties in reaching participants, this strategy may produce unbiased estimates for important health indicators. PMID- 20013147 TI - Central nervous system relapse continues to be a therapeutic challenge in extensive disease small-cell lung cancer patients with initial symptomatic brain metastases and good response to chemoradiotherapy. AB - A summary is presented of four case studies of extensive disease small-cell lung cancer patients with initial symptomatic brain metastases, who were initially successfully treated with concurrent chemoradiotherapy of the neurocranium followed by consolidation chemotherapy and thoracic chemoradiotherapy, but shortly after the initial treatment developed a sequence of central nervous system relapses despite second-line therapy. Symptoms of the effect on the central nervous system dominated the course of the cancer disease, whereas the primary tumor mass remained in complete remission in all four patients until the end of the follow-up period. PMID- 20013148 TI - Genetic polymorphisms of DNA repair gene XRCC1 and risk of uterine leiomyoma. AB - The objective was to study the relationship between the polymorphisms of the DNA repair gene XRCC1 Arg399Gln, Arg194Trp, and Arg280His uterine leiomyoma in a Chinese population. In the case-control study, we compared the XRCC1 gene polymorphism of 136 uterine leiomyoma patients and 140 healthy controls by using the polymerase chain reaction-restriction fragment length polymorphism (PCR RFLP). The results suggested that the genotype Arg/Arg of codon 280 was significantly different from its heterozygote (odds ratio [OR] = 3.633, 95% confidence interval [CI]: 2.147-6.148). In conclusion, the results suggest that polymorphism of XRCC1 Arg280His was associated with the increased risk of uterine leiomyoma in a Chinese population. PMID- 20013146 TI - Angiogenesis and expression of estrogen and progesterone receptors as predictive factors for recurrence of meningioma. AB - Meningiomas are benign tumors, with low rate of recurrence after surgery. The most important factor predicting recurrence is the extent of surgical resection; other factors have been studied with conflicting results. Angiogenesis, an important substratum for growth and spread of neoplasic cells, and the expression of estrogen and progesterone receptors (ER, PR), could play a role in the recurrence of meningioma. We evaluated 42 patients with meningioma diagnosis (confirmed by histopathology) treated exclusively by surgery between January 1995 and December 1999, and compared the recurring and non-recurring groups after a ten-year follow-up period. Recurrence was associated with several factors including vascular density (VD), cell proliferation index (CPI), ER, PR, and cyclin E (CE) tissue expression, as evaluated by immunohistochemistry. Complete surgical resection was achieved in 41% of patients. Recurrence of meningioma was found in 17 patients (40%). Median + or - standard deviation (SD) of recurrence time was 32 + or - 5 months. When recurrence versus no recurrence was compared, mean + or - SD of VD and CPI were 9 + or - 3.6 and 607.6 + or - 233 (40x/10 fields) respectively. Tissue expression was positive for ER, PR, and CE in 28, 62 and 91% of patients, respectively. The sole significant recurrence-associated factors were extent of resection (P = 0.003) and VD (P = 0.004). ER, PR, and CE tissue expression were not statistically significant. The most important factor associated with meningioma relapse was vascular density, independently of hormonal status and extent of surgical resection. Patients with a high risk of recurrence could benefit from additional treatment. PMID- 20013150 TI - Traits of separation anxiety in childhood: a retrospective study of Samoan men, women, and fa'afafine. AB - Gender Identity Disorder in Childhood (GIDC) and Separation Anxiety Disorder have been found to be co-morbid for boys in Western populations. In a Samoan cultural context, fa'afafine are defined as androphilic males (i.e., biological males who are sexually attracted to and aroused by adult males) who are effeminate or transgendered and occupy an "alternative" gender role category, which is distinct from the gender normative categories of "man" and "woman." Because some Western clinicians would likely conclude that many of the fa'afafine had GIDC, we examined whether adult fa'afafine would also recall traits indicative of elevated separation anxiety in childhood. Given this, the present study compared retrospective reports of childhood separation anxiety for Samoan men, women, and fa'afafine. Our results indicated that Samoan fa'afafine recalled significantly more separation anxiety in childhood compared to Samoan men and women. These findings, coupled with research on U.S. and Canadian boys with GIDC, suggest that separation anxiety in childhood may represent a universal pattern of psychosexual development shared by most gender-atypical, androphilic males, regardless of their cultural milieu. As such, this study adds to a growing body of literature that documents the existence of cross-culturally invariant behavioral and psychological traits in male androphiles. PMID- 20013149 TI - Adolescent emotional distress: the role of family obligations and school connectedness. AB - The current study draws upon ecodevelopmental theory to identify protective and risk factors that may influence emotional distress during adolescence. Hierarchical regression analyses were used to examine the relationship among family obligations, school connectedness and emotional distress of 4,198 (51% female) middle and high school students who were primarily (59%) European American. The overall model explained 21.1% of the variance in student emotional distress. A significant interaction effect was found indicating that school connectedness moderated the relationship between family obligations and emotional distress. Specifically, for students with low to moderate levels of family obligations, a stronger sense of school connectedness was associated with lower emotional distress. The buffering effect of school connectedness was weakened as the level of family obligations increased and completely disappeared for students who experienced high levels of family obligations. The creation of a program that takes a holistic approach, in order to curtail the levels of highly emotionally distressed adolescents, must continue to address the ever changing demands that adolescents encounter and prepare youth to deal with functioning within multiple contexts and do so while maintaining emotional well-being. PMID- 20013152 TI - Chk1-cyclin A/Cdk1 axis regulates origin firing programs in mammals. AB - DNA replication is key to ensuring the complete duplication of genomic DNA prior to mitosis and is tightly regulated by both cell cycle machinery and checkpoint signals. Regulation of the S phase program occurs at several stages, affecting origin firing, replication fork elongation, fork velocity, and fork stability, all of which are dependent on S-phase-promoting kinase activity. Somatic mammalian cells use well-established origin programs by which specific regions of the genome are replicated at precise times. However, the mechanisms by which S phase kinases regulate origin firing in mammals are largely unknown. Here, we discuss recent advances in the understanding of how S phase programs are regulated in mammals at the correct regions and at the appropriate times. PMID- 20013151 TI - Domain-wide regulation of DNA replication timing during mammalian development. AB - Studies of replication timing provide a handle into previously impenetrable higher-order levels of chromosome organization and their plasticity during development. Although mechanisms regulating replication timing are not clear, novel genome-wide studies provide a thorough survey of the extent to which replication timing is regulated during most of the early cell fate transitions in mammals, revealing coordinated changes of a defined set of 400-800 kb chromosomal segments that involve at least half the genome. Furthermore, changes in replication time are linked to changes in sub-nuclear organization and domain wide transcriptional potential, and tissue-specific replication timing profiles are conserved from mouse to human, suggesting that the program has developmental significance. Hence, these studies have provided a solid foundation for linking megabase level chromosome structure to function, and suggest a central role for replication in domain-level genome organization. PMID- 20013153 TI - Non-association between Neo-5 personality tests and multiple mini-interview. AB - Most medical schools attempt to select applicants on the basis of cognitive and non-cognitive skills. Typically, interpersonal skills are assessed by interview, though relatively few applicants make it to interview. Thus, an efficient paper and pencil test of non-cognitive skills is needed. One possibility is personality tests. Tests of the five factor model of personality, and in particular the factor of conscientiousness, has proven effective in predicting future job performance. Can it serve as a screen for admissions interviews? In particular, correlation with the multiple mini-interviews (MMI) is of interest since the latter is a well validated test of non-cognitive skills. A total of 152 applicants to Michael G. DeGroote School of Medicine at McMaster completed the Neo-5 personality test voluntarily in advance of their admissions interviews. Correlations were calculated between personality factors and grade point average (GPA), medical college admissions test (MCAT) and MMI. No statistically significant correlation was found between personality factors and cognitive (GPA, MCAT) measures. More surprisingly, no statistically significant correlation was found between personality factors, including conscientiousness, and the MMI. Personality testing is not a useful screening test for the MMI. PMID- 20013154 TI - Endothelial cell proliferation is enhanced by low dose non-thermal plasma through fibroblast growth factor-2 release. AB - Non-thermal dielectric barrier discharge plasma is being developed for a wide range of medical applications, including wound healing, blood coagulation, and malignant cell apoptosis. However, the effect of non-thermal plasma on the vasculature is unclear. Blood vessels are affected during plasma treatment of many tissues and may be an important potential target for clinical plasma therapy. Porcine aortic endothelial cells were treated in vitro with a custom non thermal plasma device. Low dose plasma (up to 30 s or 4 J cm(-2)) was relatively non-toxic to endothelial cells while treatment at longer exposures (60 s and higher or 8 J cm(-2)) led to cell death. Endothelial cells treated with plasma for 30 s demonstrated twice as much proliferation as untreated cells five days after plasma treatment. Endothelial cell release of fibroblast growth factor-2 (FGF2) peaked 3 h after plasma treatment. The plasma proliferative effect was abrogated by an FGF2 neutralizing antibody, and FGF2 release was blocked by reactive oxygen species scavengers. These data suggest that low dose non-thermal plasma enhances endothelial cell proliferation due to reactive oxygen species mediated FGF2 release. Plasma may be a novel therapy for dose-dependent promotion or inhibition of endothelial cell mediated angiogenesis. PMID- 20013155 TI - Classification of leukemia blood samples using neural networks. AB - Pattern recognition applied to blood samples for diagnosing leukemia remains an extremely difficult task which frequently leads to misclassification errors due in large part to the inherent problem of data overlap. A novel artificial neural network (ANN) algorithm is proposed for optimizing the classification of multidimensional data, focusing on acute leukemia samples. The programming tool established around the ANN architecture focuses on the classification of normal vs. abnormal blood samples, namely acute lymphocytic leukemia (ALL) and acute myeloid leukemia (AML). There were 220 blood samples considered with 60 abnormal samples and 160 normal samples. The algorithm produced very high sensitivity results that improved up to 96.67% in ALL classification with increased data set size. With this type of accuracy, this programming tool provides information to medical doctors in the form of diagnostic references for the specific disease states that are considered for this study. The results obtained prove that a neural network classifier can perform remarkably well for this type of flow cytometry data. Even more significant is the fact that experimental evaluations in the testing phase reveal that as the ALL data considered is gradually increased from small to large data sets, the more accurate are the classification results. PMID- 20013156 TI - Numerical modeling of long bone adaptation due to mechanical loading: correlation with experiments. AB - The process of external bone adaptation in cortical bone is modeled mathematically using finite element (FE) stress analysis coupled with an evolution model, in which adaptation response is triggered by mechanical stimulus represented by strain energy density. The model is applied to experiments in which a rat ulna is subjected to cyclic loading, and the results demonstrate the ability of the model to predict the bone adaptation response. The FE mesh is generated from micro-computed tomography (microCT) images of the rat ulna, and the stress analysis is carried out using boundary and loading conditions on the rat ulna obtained from the experiments [Robling, A. G., F. M. Hinant, D. B. Burr, and C. H. Turner. J. Bone Miner. Res. 17:1545-1554, 2002]. The external adaptation process is implemented in the model by moving the surface nodes of the FE mesh based on an evolution law characterized by two parameters: one that captures the rate of the adaptation process (referred to as gain); and the other characterizing the threshold value of the mechanical stimulus required for adaptation (referred to as threshold-sensitivity). A parametric study is carried out to evaluate the effect of these two parameters on the adaptation response. We show, following comparison of results from the simulations to the experimental observations of Robling et al. (J. Bone Miner. Res. 17:1545-1554, 2002), that splitting the loading cycles into different number of bouts affects the threshold sensitivity but not the rate of adaptation. We also show that the threshold sensitivity parameter can quantify the mechanosensitivity of the osteocytes. PMID- 20013158 TI - Is traditional rural lifestyle a barrier for quality of life assessment? A case study using the Short Form 36 in a rural Chinese population. AB - PURPOSE: The majority of existing quality of life measures are based on urban living environments. This study aimed at exploring the validity of using an urban lifestyle-based health questionnaire with individuals living a traditional rural lifestyle. METHODS: The Short Form-36 (SF-36) interview was administered to 1603 rural Chinese residents. Semantic ambiguity of the items was investigated using tests of internal consistency, test-retest reliability, exploratory factor analysis, and clustering and ordering of item mean scores. The self-explanations from the respondents were adopted to interpret the implications of the changes in meanings of the items. RESULTS: Cronbach's alpha reliability coefficients were high, whereas test-retest reliabilities were low. Consistent with the original factor structure, eight factors were extracted using exploratory factor analysis. However, the composition of these eight factors was not in full accordance with the priori assignment of items to scales. Seven items violated the clustering and ordering of item mean scores. The association between the identified problems in validity and the change in semantic meanings in the context of the rural lifestyle was established. CONCLUSIONS: Quality of life assessment instruments based on urban-living arrangements may not be reliably used with individuals living in rural environments. PMID- 20013157 TI - Numerical analysis of ischemia- and compression-induced injury in tissue engineered skeletal muscle constructs. AB - Pressure-related deep tissue injury may develop in skeletal muscle tissue which is subjected to prolonged compression. For early detection, it is important to understand the underlying damage processes. Gawlitta et al. [Gawlitta, D., C. W. J. Oomens, D. L. Bader, F. P. T. Baaijens, and C. V. C. Bouten. Temporal differences in the influence of ischemic factors and deformation on the metabolism of engineered skeletal muscle. J. Appl. Physiol. 103(2):464-473, 2007b] subjected tissue-engineered muscle constructs to ischemia and deformation to study their effects on viability. Contrary to previous findings, no decrease in viability was found due to compression. However, the nature of their measurement method complicated interpretation of the results, particulary when deformation was involved. Changes in the constructs were assessed by measurements in the surrounding medium. The theoretical model developed in the present study describes metabolism, diffusion, and cell death in the experiments, and accounts for reduced diffusion due to compression. It was demonstrated that the lack of effect of compression on tissue viability, as measured in the experiments, could be explained by the compression-induced decrease in diffusivity. Compression did lead to considerable cell death but this could not be measured by Gawlitta et al. [Gawlitta, D., C. W. J. Oomens, D. L. Bader, F. P. T. Baaijens, and C. V. C. Bouten. Temporal differences in the influence of ischemic factors and deformation on the metabolism of engineered skeletal muscle. J. Appl. Physiol. 103(2):464 473, 2007b] because diffusion of the cell death marker to the medium was limited. This study shows that a proper description of transport processes is essential for a correct interpretation of experiments in which indirect measurement methods are used. PMID- 20013159 TI - Musculocutaneous neuropathy: case report and discussion. AB - The musculocutaneous nerve arises from the lateral cord of the brachial plexus and contains fibers from the C5, C6, and C7 spinal nerve roots. It innervates such muscles as the biceps brachii and brachialis as well as supply branches to the skin over the lateral cubital and forearm regions via the lateral antebrachial cutaneous nerve. Musculocutaneous neuropathy can arise from exercise, participating in sports, strenuous activity, cast placement, trauma, and surgery in addition to other less understood causes such as Parsonage Turner syndrome. We present the case of a 55-year-old female who complained of numbness, weakness, and pain throughout the arm starting 1 day following a surgical procedure. Electrodiagnostic testing revealed a musculocutaneous neuropathy with significant axonal injury. Symptoms of musculocutaneous neuropathy may be similar to cervical spinal nerve root impingement or brachial plexus lesions. Therefore, magnetic resonance imaging and electrodiagnostic studies may be useful in differentiating between these conditions. Once the diagnosis of musculocutaneous neuropathy has been made, treatments include relative rest, nonsteroidal anti inflammatory drugs, splinting, physical therapy, and surgical decompression in cases that do not respond to conservative management. PMID- 20013160 TI - Chromium improves protein deposition through regulating the mRNA levels of IGF-1, IGF-1R, and Ub in rat skeletal muscle cells. AB - The aim of this study was to evaluate the impact of three different chromium forms--chromic chloride (CrCl3), chromium picolinate (CrPic), and a newly synthesized complex of chromium chelated with small peptides (CrSP)--on protein metabolism in vitro. In cultured skeletal muscle cells, CrSP was able to increase the basal and insulin-stimulated levels of protein deposition in skeletal muscles cells. CrCl3 and CrPic augmented insulin-stimulated protein synthesis. At the molecular level, insulin significantly increased the mRNA levels of insulin-like growth factor 1 and insulin-like growth factor 1 receptor. These impacts could be enhanced by the addition of chromium, especially CrSP. The mRNA levels of ubiquitin were significantly reduced when cells were cultured with chromium or/and insulin. Assuming that the mRNA level increase or decrease results in increased or decreased levels of these proteins, chromium would improve protein anabolism and reduce protein catabolism and then prove protein deposition in rat skeletal muscle cells. PMID- 20013161 TI - Correlation between dietary zinc intakes and its serum levels with depression scales in young female students. AB - It has been suggested that mood disorders and depressive status may be accompanied by lowered zinc status in the body, and adequate consumption of zinc increases a general perceived well-being. The main objective of this study was to assess the correlation between serum zinc concentrations and dietary zinc intakes with depression scores in university female students. In the first phase, Beck's depression questionnaire was applied in a random sampling of 308 selected 20-25 year-old female students (one third of total students in Ahvaz Jondi-Shapour University of Medical Sciences Golestan dormitories) to assess the major depressive disorder (MDD) scales. Then, in the second phase, 23 students who identified as having moderate and severe depression were selected as the case group, and 23 healthy age matched were chosen as the controls. Each of them completed a 12-item semiquantitative food frequency questionnaire containing the main food sources of zinc in the usual dietary patterns and also a 24-h food recall questionnaire to assure the daily zinc intakes. Daily zinc intakes were obtained by multiplying each portion size by its zinc content using food tables. A 5-ml blood sample was taken for further serum zinc status using flame atomic absorption spectrophotometry technique. Pearson's r was used to show the correlation between quantitative variables. Both daily zinc intake and serum zinc concentrations of MDD group were about two thirds of healthy index (p < 0.01). Depressed individuals used to eat lower servings of red meats and chicken as the main food sources of zinc in students' usual diets (p < 0.001). Consumption of other foods as the sources of zinc was not significantly different in two groups. A linear significant correlation between dietary zinc intakes and its serum levels was seen in samples (r = 0.62; p < 0.001) and MDD students (r = 0.55; p < 0.001). There was a linear inverse correlation between Beck questionnaire scores and serum zinc concentrations in all of the investigated students(r = -0.65; p < 0.001) and MDD girls (r = -0.71; p < 0.001). Beck questionnaire scores and diatary zinc intakes were also inversly correlated (r = -0.58; p < 0.001). However, no statistical correlation was seen between these two variables in MDD cases. In depressed female students, dietary zinc intake is correlated to its serum concentrations; however, the serum zinc levels are inversely correlated to depression scales. Consumption of the main dietary sources of zinc such as red meats and chicken should be encouraged in young depressed girls. PMID- 20013162 TI - 1H, 13C and 15N resonance assignments of the Calmodulin-Munc13-1 peptide complex. AB - Ca(2+)-Calmodulin binding to the variable N-terminal region of the diacylglycerol/phorbol ester-binding UNC13/Munc13 family of proteins modulates the short-term synaptic plasticity characteristics in neurons. Here, we report the sequential backbone and side chain resonance assignment of the Ca(2+) Calmodulin/Munc13-1(458-492) peptide complex at pH 6.8 and 35 degrees C (BMRB No. 15470). PMID- 20013164 TI - Cartilage degeneration is associated with augmented chemically-induced joint pain in rats: a pilot study. AB - BACKGROUND: Osteoarthritis arising from cartilage degeneration is the most common cause of joint pain. However, the relationship between joint pain and cartilage degeneration is not well understood. QUESTIONS/PURPOSES: We asked whether the inflammatory mediators participate in the joint pain in the presence of cartilage degeneration. METHODS: We observed electromyographic responses of hindlimb flexors to four inflammatory mediators (bradykinin, ATP, acetylcholine, and serotonin) injected in normal rat knees and in those with monosodium iodoacetate (MIA)-induced arthritis. RESULTS: Joint cartilage of all the rats with MIA induced arthritis histologically showed severe degeneration. We observed greater magnitude and longer duration responses in the MIA-induced arthritis than normal joints with all four mediators. CONCLUSIONS: The data suggested nociceptors in osteoarthritic joints are more sensitive to inflammatory mediators than in normal joints. Such nociceptive sensitization to inflammatory mediators may participate in the joint pain in osteoarthritis. PMID- 20013166 TI - Erythroderma related to the administration of 99mTc-sestamibi: the first report. PMID- 20013167 TI - Myocardial imaging with 99mTc-Tetrofosmin: Influence of post-stress acquisition time, regional radiotracer uptake, and wall motion abnormalities on the clinical result. AB - BACKGROUND: We previously demonstrated that early (15', T1) post-stress myocardial imaging with Tetrofosmin could be more accurate than standard acquisitions (45', T2) in identifying coronary artery disease. METHODS AND RESULTS: To clarify this phenomenon, 120 subjects (age 61 +/- 10 years) with both T1 and T2 scans were divided into Group 1 (53/120 pts) with more ischemia at T1 vs T2 imaging (T1-T2SDS > or = 3); Group 2 (67/120 pts) with similar results (T1 T2SDS < or = 2). Myocardial areas were categorized as control nonischemic, ischemic, and scarred on the basis of perfusion/contraction properties and coronary anatomy. In each area, regional myocardial count statistic and semiquantitative wall motion/thickening values were obtained. Analysis of T1 and T2 post-stress myocardial counts demonstrated a significant Tetrofosmin wash-out rate that was higher in Group 1 control nonischemic regions (15 +/- 8% vs 13.6 +/ 9.6%, P < .02), significantly lower in Group 1 ischemic regions (7 +/- 10% vs 12.2 +/- 9.5%, P < .0001), and comparable between scarred areas of the two groups (P = NS). Delta post-stress wall thickening (T1-T2) was lower in Group 1 ischemic regions (-4.5 +/- 9.15% vs -1.90 +/- 7.0%, P < .001) and comparable in both control nonischemic and scarred areas of the two groups (P = NS). CONCLUSION: The clinical result of Tetrofosmin gated-SPECT can be influenced by the post-stress acquisition time because of ischemic-induced regional wall thickening abnormalities and the existence of a differential radiotracer myocardial wash out. PMID- 20013165 TI - Impact of carbohydrate restriction with and without fatty acid loading on myocardial 18F-FDG uptake during PET: A randomized controlled trial. AB - BACKGROUND: Low-carbohydrate (LC) and high-fat, low-carbohydrate (HFLC) dietary preparations may enhance (18)F-FDG-PET-based imaging of small, inflamed structures near the heart by suppressing myocardial FDG signal. We compared myocardial (18)F-FDG uptake in patients randomized to LC, HFLC, and unrestricted (UR) preparations prior to (18)F-FDG-PET. METHODS AND RESULTS: We randomized 63 outpatients referred for oncologic (18)F-FDG-PET to LC, HFLC, or UR dietary preparations (1:1:1 allocation) starting the evening before PET. After eating dinner according to instructions, UR and LC patients fasted until FDG injection (mean time 745 minutes for UR, 899 minutes for LC), and HFLC patients drank a fatty drink 60-70 minutes prior to FDG injection. Attenuation-corrected PET imaging was performed 60 minutes after FDG administration. Maximal myocardial standard uptake values (MyoSUV(max)) were systematically measured in axial view and compared between the three groups. Using UR patients as reference, mean MyoSUV(max) was lower in LC patients (3.3 +/- 2.7 vs 6.2 +/- 5.2, P = .03) but not in HFLC patients (5.5 +/- 4.2, P = .63). Ratios of MyoSUV(max) to liver SUV(max), calculated to control for background uptake, were not significantly different amongst the groups (1.9 +/- 2.1 LC, 2.6 +/- 2.3 HFLC, 3.6 +/- 3.5 UR). CONCLUSION: In this small randomized controlled trial using UR diet as reference, LC dietary preparation followed by extended fasting resulted in significant myocardial uptake suppression. PMID- 20013170 TI - In vivo measurement of tissue oxygen using electron paramagnetic resonance spectroscopy with oxygen-sensitive paramagnetic particle, lithium phthalocyanine. AB - The partial pressure of oxygen (pO(2)) plays a determining role in the energy metabolism of aerobic cells. However, low pO(2) level induces pathophysiological conditions such as tumor hypoxia, ischemia or reperfusion injury, and delayed/altered wound healing. Especially, pO(2) level in the tumor is known to be related to tumor progression and effectiveness of radiotherapy. To monitor the pO(2) levels in vivo, continuous wave (CW) and time-domain (TD) electron paramagnetic resonance (EPR) spectroscopy method was used, in which surface coil resonator and Lithium phthalocyanine (LiPc) as oxygen sensor were crucial. Once LiPc particles are embedded in a desired location of organ/tissue, the pO(2) level can be monitored repeatedly and non-invasively. This method is based on the effect of oxygen concentration on the EPR spectra of LiPc which offers several advantages as follows: (1) high sensitivity, (2) minimum invasiveness, (3) repeated measurements, (4) absence of toxicity (non-toxic), and (5) measurement in a local region of the tissue with embedded LiPc. Therefore, in this chapter, we describe the method using CW and TD EPR spectroscopy with oxygen-sensitive particle, LiPc, for in vivo monitoring of oxygen. PMID- 20013171 TI - Measurement of plasma nitrite by chemiluminescence. AB - Studies have demonstrated that plasma nitrite (N(O-)(2)) reflects endothelial nitric oxide (NO) production. In addition, N(O-)(2) has been shown to have biological activities associated with its reduction to NO in blood and tissues. Therefore, determination of plasma N(O-)(2) has been proposed as a prognostic marker for cardiovascular diseases. Typical concentrations of N(O-)(2) in the plasma are in the nanomolar range and determination of this N(O-)(2) poses a challenge in terms of both sensitivity and specificity. Thus, a highly sensitive, chemiluminescence method that is based on the reduction of N(O-)(2) by potassium iodide and iodine is being used to determine the nitrite in biological fluids. This method has the sensitivity, but also measures other nitric oxide species such as S-nitrosothiols and N-nitrosamines. We, therefore, developed an alternative method based on the reduction of N(O-)(2) by ascorbic acid in strongly acidic media. As part of the methodology, glacial acetic acid and ascorbic acid are introduced into the purge vessel of the NO analyzer. Samples containing N(O-)(2) are injected into the purge vessel and the chemiluminescence signals generated as a result of the formation of NO are then measured. We find that under these conditions N(O-)(2) is stoichiometrically reduced to NO. Other traditional NO-generating species, such as S-nitrosothiols, N-nitrosamines, nitrated lipids, and nitrated proteins, did not interfere in the determination of plasma N(O-)(2). Using the present method, plasma N(O-)(2) in fasting human subjects has been determined to be in the range of 56-210 nM (mean +/- SD = 110 +/- 36 nM; n = 8). PMID- 20013172 TI - Determination of glutathione, mitochondrial transmembrane potential, and cytotoxicity in H9c2 cardiomyoblasts exposed to reactive oxygen and nitrogen species. AB - Quantitative measurement of cellular oxidative stress (COS) and cytotoxicity are important to establish their significance in pathophysiologic conditions and disease states. So far, ample methods have been described to determine these processes based on spectrophotometric analysis. The application of simple, rapid, and sensitive fluorescence methods to determine the cytotoxicity and COS is described in the present chapter. Murine H9c2 cells were exposed to various free radical and non-free radical oxidants through use of diethylamine NONOate, 3 morpholinosydnonimine (SIN-1), and a synthetic preparation of peroxynitrite (PN). The viability of control and the treated H9c2 cells was measured based on the reduction of resazurin to resorufin which generates a fluorescent signal. The mitochondrial membrane potential was quantified by determining the cellular uptake of a fluorescent dye, (5,5('),6,6(')-tetrachloro-1,1(')-3,3(') tetraethylbenzimidazolcarbocyanine iodide (JC-1)) and its segregation in the mitochondrial fraction. The intracellular GSH was determined by assaying the glutathione-S-transferase (GST)-catalyzed conjugation of GSH to monochlorobimane. This chapter describes the feasibility and potential of the above-described fluorescence approach as simple alternative methods to determine reactive oxygen and nitrogen species-induced cytotoxicity and oxidative stress using H9c2 cardiomyoblasts as a model system. PMID- 20013169 TI - In vivo imaging of free radicals and oxygen. AB - Free radicals are highly reactive compounds that play an essential role in many biological processes, both beneficial and deleterious. Detection and quantification of these species is critical to develop a better understanding of normal and pathophysiological functions at the cellular and tissue levels. Electron paramagnetic resonance (EPR) spectroscopy is the technique most commonly used for this purpose through the detection of exogenous probes or spin traps that interact with the free radical species of interest. Over the past several years, the spatial and temporal distribution of free radicals within cells and tissues has been of particular interest. This chapter briefly explains the principles and challenges in the use of EPR for biological samples and introduces the concept of EPR for free radical imaging purposes. In addition, specific examples are given for the use of EPR imaging in four principal areas: free radical probes, nitric oxide (NO), redox state, and oxygen (O(2)) concentration. PMID- 20013173 TI - Phenolic acids and flavonoids: occurrence and analytical methods. AB - Phenolics are structurally assorted and are generally part of a complex mixture isolated from plant and biological origin matrices. A wide gamut of natural products have been the focus of main study for phenolic compounds while urine and blood are the two main biological fluids that have been analyzed for metabolism studies. Traditional and more advanced techniques have come to prominence for sample preparation, detection, and identification. This review is devoted to a short discussion of the occurrence of phenolic acids and flavonoids, their role in human health, and focuses on a detailed presentation of the analytical methods, concluding with the advantages of analytical methods employed so far and prospects. Strategies and practical aspects for the determination of phenolic acids and flavonoids in biological fluids, beverages, plants, and food are reported. Novel and past applications are provided with significant treatment and detection-related developments on the basis of the employment of separation and non-separation analytical techniques. PMID- 20013174 TI - Design, synthesis, and action of antiatherogenic antioxidants. AB - Ample evidence supports the critical role of oxidized low-density lipoprotein (ox LDL) in initiation and progression of atherosclerosis. Oxidation of LDL is a complex process involving several steps (processes) of reactions such as initiation and propagation. Both proteins and lipids in LDL undergo free radical mediated oxidations leading to the formation of ox-LDL that plays a pivotal role in atherosclerosis. Antioxidants of various types (both aqueous and lipophilic) either arrest or retard the oxidation of LDL at various steps of the oxidation process (e.g., initiation or propagation). Certain lipophilic antioxidants act as the chain-terminating antioxidants leading to the inhibition of LDL oxidation. The current chapter describes the designing and efficacy of two novel lipophilic antioxidants (benzofuranol, BO-653 and aniline, BO-313) in inhibiting the LDL oxidation and atherogenesis in experimental animal model. Furthermore, the characteristics of an effective antioxidant to inhibit LDL oxidation and atherogenesis which dictates the designing of the antioxidant drug and its mechanism(s) of antiatherogenic action are discussed. PMID- 20013175 TI - Preparation of drug-loaded polymeric nanoparticles and evaluation of the antioxidant activity against lipid peroxidation. AB - Antioxidants have been found to be effective as prophylatic and therapeutic agents for different diseases such as diabetes, cancer, and neurodegenerative disorders. However, antioxidant substances can present poor solubility in water, inefficient permeability, gastrointestinal degradation, first-pass effect, and/or instability during storage. These drawbacks can be potentially circumvented by encapsulating the susceptible antioxidants. Polymeric nanoparticles (nanocapsules or nanospheres) have been used to improve the drug efficacy and release. Our group has shown that the in vitro antioxidant effect of melatonin against lipid peroxidation in microsomes and liposomes can be improved by encapsulation of the antioxidant drug in polymeric nanoparticles. PMID- 20013176 TI - Nanoparticle and iron chelators as a potential novel Alzheimer therapy. AB - Current therapies for Alzheimer disease (AD) such as the acetylcholinesterase inhibitors and the latest NMDA receptor inhibitor, Namenda, provide moderate symptomatic delay at various stages of the disease, but do not arrest the disease progression or bring in meaningful remission. New approaches to the disease management are urgently needed. Although the etiology of AD is largely unknown, oxidative damage mediated by metals is likely a significant contributor since metals such as iron, aluminum, zinc, and copper are dysregulated and/or increased in AD brain tissue and create a pro-oxidative environment. This role of metal ion induced free radical formation in AD makes chelation therapy an attractive means of dampening the oxidative stress burden in neurons. The chelator desferrioxamine, FDA approved for iron overload, has shown some benefit in AD, but like many chelators, it has a host of adverse effects and substantial obstacles for tissue-specific targeting. Other chelators are under development and have shown various strengths and weaknesses. Here, we propose a novel system of chelation therapy through the use of nanoparticles. Nanoparticles conjugated to chelators show unique ability to cross the blood-brain barrier (BBB), chelate metals, and exit through the BBB with their corresponding complexed metal ions. This method may provide a safer and more effective means of reducing the metal load in neural tissue, thus attenuating the harmful effects of oxidative damage and its sequelae. Experimental procedures are presented in this chapter. PMID- 20013177 TI - Synthesis and characterization of polymer nanocarriers for the targeted delivery of therapeutic enzymes. AB - Protein drugs, such as recombinant enzymes useful for detoxification and replacement therapies, have extraordinary specificity and potency. However, inherently inadequate delivery to target sites and rapid inactivation limit their medical utility. Using chaperone polymeric particles designed within an injectible size range (sub-micron) may help solve these shortcomings. Such nanocarriers would (i) prevent premature inactivation of encapsulated therapeutic protein cargoes, (ii) provide a carrier that can be surface decorated by targeting ligands, and (iii) optimize sub-cellular localization of the drug. This chapter describes the techniques successfully employed for the preparation of polymer nanocarriers (PNC) loaded with the antioxidant enzyme, catalase, and targeted to endothelial cells. Methods of PNC synthesis, loading with catalase, characterization, coupling of a targeting moiety, and in vitro testing of the enzymatic and targeting activities are provided here. Advantages and disadvantages of specific designs are discussed. Due to the modular nature of the targeting methodology employed, it is believed that these protocols will provide a solid foundation for the formulation of a wide variety of enzymatic drug targeting strategies. PMID- 20013179 TI - The generation of stable oxidative stress-resistant phenotypes in Chinese hamster fibroblasts chronically exposed to hydrogen peroxide or hyperoxia. AB - With the recognition that chronic exposure to oxidative stress occurs in many disease states and can be deleterious to the human health, great interest has emerged in understanding the mechanisms by which mammalian cells develop resistance to chronic oxidative stress. In order to study the mechanisms of development of resistance to chronic oxidative stress, a model system where Chinese hamster fibroblasts (HA1) are chronically exposed to progressively increasing concentrations of H(2)O(2) (50-800 muM) or hyperoxia (80-95% O(2)) has been developed. Following >200 d of exposure to H(2)O(2) (or 18 months of exposure to hyperoxia), the cells developed stable H(2)O(2)-resistant (or O(2) resistant) phenotypes that are characterized by increases in total glutathione, antioxidant enzyme activity, heme oxygenase activity, stress protein gene expression, DNA repair pathways, and resistance to a wide variety of other toxic stress known to cause oxidant injury. In addition, these oxidant-resistant cells exhibited amplification of the gene for catalase and constitutively elevated AP-1 DNA binding activity. Further, beyond 240 d genomic instability as evidenced by chromosomal rearrangements and alterations in ploidy was stably maintained upon removal from the chronic oxidative stress conditions. These results demonstrate the capability of mammalian cells to develop stable oxidative stress-resistant phenotypes in response to both exogenous (H(2)O(2)) as well as endogenous (95% O(2)) oxidative stress. The understanding of mechanisms of resistance to oxidative stress and its possible relevance in various disease states are discussed. PMID- 20013178 TI - Assessment of antioxidant activity of eugenol in vitro and in vivo. AB - Reactive oxygen species are implicated in many human diseases and aging process. Much of the evidence is based on experimental data indicating increasing rates of lipid peroxidation in disease states and the ameliorating effects of antioxidants. It is becoming increasingly evident that the natural antioxidants, which have phenolic structure, play an important role in protecting the tissues against free radical damage. Eugenol (4-allyl-2 methoxyphenol) is one such naturally occurring phenolic compound. The antioxidant activity of eugenol was evaluated by the extent of protection offered against free radical-mediated lipid peroxidation using both in vitro and in vivo studies. The in vitro lipid peroxidation was induced in mitochondria by (Fe(II)-ascorbate) or (Fe(II) + H(2)O(2)). The lipid peroxidation was assessed colorimetrically by measuring the formation of thiobarbituric acid reactive substances (TBARS) following the reaction of oxidized lipids with TBA. Eugenol completely inhibited both iron and Fenton reagent-mediated lipid peroxidation. The inhibitory activity of eugenol was about fivefold higher than that observed for alpha-tocopherol and about tenfold less than that observed for BHT. The in vivo lipid peroxidation-mediated liver damage was induced by administration of CCl(4) to rats. Eugenol significantly inhibited the rise in SGOT activity and cell necrosis without protecting the endoplasmic reticulum (ER) damage as assessed by its failure to prevent a decrease in cytochrome p450 and G-6-phosphatase activity. The protective action of eugenol has been found to be due to interception of secondary radicals derived from ER lipids rather than interfering with primary radicals of CCl(4) (CCl(3)/CCl(3)OO). PMID- 20013181 TI - Superoxide dismutase - a target for gene therapeutic approach to reduce oxidative stress in erectile dysfunction. AB - Erectile dysfunction (ED) is defined as the inability to attain and/or maintain penile erection sufficient for satisfactory sexual performance. Oxidative stress has been demonstrated to be involved in the pathophysiology of age- or diabetes related ED. Superoxide dismutase (SOD), an antioxidant enzyme catalyzing the conversion of superoxide anion (O(2) (-)) to hydrogen peroxide (H(2)O(2)) and molecular oxygen (O(2)), is a promising therapeutic target for ED. In vivo gene therapy and adult stem cell-based ex vivo gene therapy are two attractive current gene therapies for the treatment of ED. In this chapter we describe the use of two potent gene transfer techniques to deliver the therapeutic gene extracellular superoxide dismutase (ecSOD) into the penis of aged or diabetic rats for therapy of ED: adenoviral-mediated intracavernosal ecSOD gene transfer for gene therapy of ED and ecSOD gene-modified marrow stromal cells, also known as mesenchymal stem cells, based stem cell and gene therapy. PMID- 20013180 TI - A simple method for effective and safe removal of membrane cholesterol from lipid rafts in vascular endothelial cells: implications in oxidant-mediated lipid signaling. AB - Lipid raft-associated cholesterol has been identified as a pivotal player among membrane lipids in regulating cellular functions. Cholesterol of the vascular endothelial cell (EC) membranes is also being recognized as an important element in the vascular EC signaling. However, methods utilized in studying the important role of lipid raft-associated cholesterol in cell signaling involve removal of the raft cholesterol with the aid of chemical agents called cyclodextrins. Caution should be exercised in using cyclodextrins to remove the cellular lipid raft-associated cholesterol as the cyclodextrins cause adverse effects on cells such as loss of cell viability or induction of cytotoxicity. Therefore, the choice of a cyclodextrin to remove the cellular lipid raft-associated cholesterol is extremely important in order to ensure effective and safe removal of cholesterol from the cellular lipid rafts. In order to achieve this, here, we have selected the bovine pulmonary artery endothelial cells (BPAECs) and subjected them to the removal of cholesterol using two different beta cyclodextrin compounds, methyl-beta-cyclodextrin (MbetaCD) and hydroxypropyl-beta cyclodextrin (HPCD). Phospholipase D (PLD), which generates one of the most potent bioactive lipid signal mediators (phosphatidic acid), is activated by oxidants. Therefore, we examined the effects of cholesterol removal by utilizing our current methods on the hydrogen peroxide (H(2)O(2))-activated PLD in BPAECs. Differences in the loss of cholesterol and the resulting effects on the cell membrane, cell viability, morphology, and the extent of oxidant-induced PLD activation were determined. The results revealed that both MbetaCD and HPCD caused loss of cholesterol, loss of cell viability, and altered cell morphology in the chosen EC system. It was also determined that the HPCD compound caused far less extensive damage to the cells than the MbetaCD, therefore making the HPCD compound a safer tool for EC cholesterol removal. PMID- 20013183 TI - Biomarkers of oxidative stress: methods and measures of oxidative DNA damage (COMET assay) and telomere shortening. AB - Oxidative stress is fast becoming the nutritional and medical buzzword for the twenty-first century. The theoretical importance of oxidative stress in diabetes is highlighted by its potential double impact on metabolic dysfunction on one hand and the vascular system on the other hand. The new concept of oxidative stress, being an important trigger in the onset and progression of diabetes and its complications, emphasizes the need for measurement of markers of oxidation to assess the degree of oxidative stress. While we have been routinely measuring biomarkers in our molecular epidemiology projects, here we discuss the utility of two assays, (a) DNA damage assessment by COMET measurement and (b) telomere length measurement. As DNA damage is efficiently repaired by cellular enzymes, its measurement gives a snapshot view of the level of oxidative stress. The protocol allows for measurement of oxidative DNA damage (FPG-sensitive DNA strand breaks). Telomere length measured by Southern blotting technique allows one to estimate the chronic burden of oxidative stress at the molecular level and is now considered as biomarker of biological aging. PMID- 20013184 TI - Simultaneous analysis of expression of multiple redox-sensitive and apoptotic genes in hypothalamic neurons exposed to cholesterol secoaldehyde. AB - Oxidative stress and apoptotic cell death are two important processes that occur under several disease states and in conditions of toxicant insult. Traditionally, investigators have chosen a variety of analytical methods to detect and/or quantify oxidative stress and apoptosis. The approach has proven less satisfying, however, when applied to complex systems with many unknown influences. Such areas of study could benefit from the development and application of new and more powerful analytical tools. Microarray-based approach has been developed for analyzing various cellular phenomena at the level of gene expression. These gene arrays are hybridization chips that are capable of simultaneous analysis of the expression of thousands of genes. Often, this approach warrants examining a multitude of unrelated genes which can greatly impede the interpretation of results. The real-time RT-PCR-based methodology presented here allows simultaneous detection and analysis of as many as 84 well-characterized genes associated with either oxidative stress or apoptosis in hypothalamic neuronal cells exposed to cholesterol secoaldehyde, an "ozone-/singlet oxygen-specific" oxidation product of cholesterol that has been shown to be present at the inflammatory sites including the arterial plaque and the brain specimens of patients with Alzheimer's disease. This pathway-specific analysis of the expression of the well-defined chosen set of genes offers ways of convenient and reliable interpretation of results that often corroborate well with the results obtained from other standard biochemical analytical approaches. PMID- 20013185 TI - Redox homeostasis and cellular stress response in aging and neurodegeneration. AB - Decreased expression and/or activity of antioxidant proteins leads to oxidative stress, accelerated aging, and neurodegeneration. While overwhelming levels and uncontrolled/dysregulated actions of reactive oxygen species (ROS) lead to deleterious effects, tighter regulation of those plays an important role in cell signaling. Mutations causing protein misfolding and the overload of toxic products derived from the free radical oxidation of polyunsaturated fatty acids, cholesterol, and glucose contribute to the disruption of the cellular redox homeostasis. Collectively or individually, these effects create pro-oxidant conditions in cells. Oxidative stress can induce neuronal damage, modulate intracellular signaling, and can ultimately lead to neuronal death by apoptosis or necrosis. Emerging evidence indicates that homocysteine (Hcy), a non-protein amino acid naturally present in the plasma, is implicated as a risk factor for numerous diseases. In particular, increased levels of circulating Hcy have been recognized as an independent risk factor for the development of vascular disease(s). Recent findings emphasize a relationship between elevated Hcy levels and neurodegeneration, which can be observed in Alzheimer's and Parkinson's diseases. An integrated response exists in the brain to detect and control diverse forms of stress. This is accomplished by a complex network of the so called longevity assurance processes, which are controlled by several genes termed "vitagenes." Among these, the heat-shock proteins (HSPs) form a highly conserved system that is responsible for the preservation and repair of the correct protein conformation. Recent studies have shown that the heat-shock response (HSR) contributes to cytoprotection in a number of human diseases including inflammation, cancer, aging, and neurodegenerative disorders. Given the broad cytoprotective properties of the HSR, interest mounts currently among investigators toward discovering and developing pharmacological agents capable of inducing HSR. L: -Acetylcarnitine (LAC) is proposed as a therapeutic agent for several neurodegenerative disorders and also current evidence suggests that the compound may play a critical role in the modulation of cellular stress response in health and disease conditions. Here, we review the emerging salient concepts highlighting the pathways of neurodegeneration and the role of LAC in modulating the redox-dependent mechanisms responsible for the upregulation of vitagenes in brain that leads to the enhancement of stress tolerance in brain. PMID- 20013182 TI - Assessing the reductive capacity of cells by measuring the recycling of ascorbic and lipoic acids. AB - Most mammalian cells cannot synthesize vitamin C, or ascorbic acid, and thus must have efficient mechanisms for its intracellular recycling. Ascorbate can be recycled from both its oxidized forms using electrons from several intracellular reducing co-factors, including GSH and the reduced pyridine nucleotides. Methods have been developed to assess the ability of intact cells to recycle ascorbate, which include assay of extracellular ferricyanide reduction and measurement of the ability of the cells to reduce dehydroascorbic acid to ascorbate. Lipoic acid, a disulfide containing medium chain fatty acid, is also taken up by cells and reduced to dihydrolipoic acid, which can be measured upon efflux from the cells using Ellman's reagent. Together, these assays provide an estimate of the ability of different cell types to recycle ascorbate and to generate intracellular reducing equivalents required to maintain the redox status of the cells. PMID- 20013188 TI - Analysis of urinary 8-oxo-7,8-dihydro-2'-deoxyguanosine by liquid chromatography tandem mass spectrometry. AB - The ability to non-invasively monitor DNA oxidation and its repair has significant utility in large-scale, population-based studies. Such studies could include assessments of the efficacy of antioxidant intervention strategies, pathological roles of DNA oxidation in various disease states and population or inter-individual differences in antioxidant defence and DNA repair. The analysis of urine, or indeed any extracellular matrix, for 8-oxo-7,8-dihydro-2' deoxyguanosine (8-oxodG), using chromatographic or immunoassay procedures, is by far the most popular method to non-invasively assess oxidative insult to the genome. The actual biological significance of the presence of extracellular 8 oxodG is still a subject for debate however. Studies are slowly ruling out confounding factors such as diet and cell turnover, which would leave endogenous processes, notably repair, as the sole source of extracellular 8-oxodG. The method described herein exploits the non-invasive properties of urine sampling, coupled with efficient extraction of 8-oxodG by a validated solid-phase extraction procedure. Subsequent analysis by liquid chromatography-tandem mass spectrometry has the advantages of sensitivity, internal standardisation and robust peak identification. PMID- 20013186 TI - Gene therapy techniques for the delivery of endothelial nitric oxide synthase to the lung for pulmonary hypertension. AB - Pulmonary hypertension (PH) is a serious, often fatal disease characterized by remodeling of the pulmonary vascular bed, increased pulmonary arterial pressure, and right heart failure. The increased vascular resistance in the pulmonary circulation is due to structural changes and increased vasoconstrictor tone. Although current therapies have prolonged survival, the long-term outcome is not favorable. Nitric oxide (NO) is synthesized by endothelial nitric oxide synthase (eNOS) and is important in regulating vascular resistance and in vascular remodeling in the lung. NO deficiency due to endothelial dysfunction plays an important role in the pathogenesis of PH. Therefore, local eNOS gene delivery to the lung is a promising approach for the treatment of PH. Adenoviral-mediated in vivo gene therapy and adult stem cell-based ex vivo gene therapy are two attractive current gene therapies for the treatment of cardiovascular and pulmonary diseases. In this chapter we describe the use of two gene transfer techniques, i.e., adenoviral gene transfer of eNOS and eNOS gene-modified rat marrow stromal cells, for eNOS gene delivery to the lung of laboratory animals for the treatment of PH. PMID- 20013187 TI - A general method for quantifying sequence effects on nucleobase oxidation in DNA. AB - Oxidative damage to DNA has long been associated with aging and disease, with guanine serving as the primary target for oxidation owing to its low ionization potential. Emerging evidence points to a critical role for sequence context as a determinant of the guanine ionization potential and the associated chemical reactivity of the guanine, as well as the spectrum of damage products that arise from oxidation. Recent studies also suggest that the generally accepted model of oxidation hotspots in runs of guanine bases may not hold for biologically relevant oxidants. One of the primary methods used to address these important problems of sequence context utilizes gel electrophoresis to identify the location and quantity of base damage arising in model oligonucleotides. However, this approach has limited study to those agents that produce few strand breaks arising from deoxyribose oxidation, while ionizing radiation, Fenton chemistry and other biologically relevant oxidants produce sizeable proportions of both base and sugar damage. To this end, we have developed a universal method to quantify sequence context effects on nucleobase damage without interference by strand breaks from deoxyribose oxidation. PMID- 20013189 TI - Oxidative lipidomics of apoptosis: quantitative assessment of phospholipid hydroperoxides in cells and tissues. AB - Oxidized phospholipids play essential roles in execution of mitochondrial stage of apoptosis and clearance of apoptotic cells. The identification and quantification of oxidized phospholipids generated during apoptosis can be successfully achieved by oxidative lipidomics. With this approach, diverse molecular species of phospholipids and their hydroperoxides are identified and characterized by soft-ionization mass-spectrometry techniques such as electrospray ionization (ESI). Quantitative assessment of lipid hydroperoxides is performed by fluorescence HPLC-based protocol. The protocol is based on separation of phospholipids using two-dimensional-high-performance thin-layer chromatography (2-D-HPTLC). Phospholipids are hydrolyzed using phospholipase A(2). The fatty acid hydroperoxides (FA-OOH) released is quantified by a fluorometric assay using Amplex red reagent and microperoxidase-11 (MP-11). Detection limit of this protocol is 1-2 pmol of lipid hydroperoxides. Lipid arrays vs. oxidized lipid arrays can be performed by comparing the abundance of phospholipids with the abundance of oxidized phospholipids. Using oxidative lipidomics approach we show that the pattern of phospholipid oxidation during apoptosis is nonrandom and does not follow their abundance in several types of cells undergoing apoptosis and a variety of disease states. This has important implications for evaluation of apoptosis in vivo. The anionic phospholipids, cardiolipin (CL) and phosphatidylserine (PS), are the preferred peroxidation substrates. PMID- 20013190 TI - Simultaneous analysis of multiple lipid oxidation products in vivo by liquid chromatographic-mass spectrometry (LC-MS). AB - Free radical-induced oxidation of polyunsaturated fatty acid (PUFAs) has been linked to a number of human diseases including atherosclerosis and neurodegenerative disorders. Oxidation of PUFAs generates hydroperoxides and cyclic peroxides that are reduced to lipid alcohol, such as hydroxyeicosatetraenoic acid (HETEs), and isoprostanes (IsoPs) respectively. The IsoPs are isomers of prostaglandins that are generated from autoxidation of arachidonic acid (C20:4). Quantification of F(2)-IsoPs has been regarded as the "gold standard" to assess oxidative stress status in various human diseases. We herein report the protocol of analyzing HETEs and F(2)-IsoPs using a triple quadrupole mass spectrometer coupled to reverse phase liquid chromatography. The selected reaction monitoring (SRM) mode selects the parent ion of interest in the first Quad (m/z 319 for HETE and m/z 353 for F(2)-IsoPs) and fragments it in the second while an ion characteristic of the analyte of interest is monitored in the third Quad. This highly selective technique permits the simultaneous analysis of multiple oxidation products such as the HETEs and F(2)-IsoPs. This LC-MS technique can be applied to study the free radical oxidation mechanism in vitro and assess the oxidative stress status in biological tissues and fluids. PMID- 20013191 TI - Lipoxygenase-catalyzed phospholipid peroxidation: preparation, purification, and characterization of phosphatidylinositol peroxides. AB - The importance of understanding the mechanisms of modulation of cellular signaling cascades by the peroxidized membrane phospholipids (PLs) is well recognized. The enzyme-catalyzed peroxidation of PLs, as opposed to their oxidation by air and metal catalysis, is well controlled and rapid and yields well-defined PL peroxides which are highly desirable for biological studies. Therefore, here, we chose bovine liver phosphatidylinositol (PI), a crucial membrane PL which acts as the substrate for phospholipase C in cellular signal transduction, as a model membrane PL. We successfully generated the PI peroxides with soybean type-I lipoxygenase (LOX) in the presence of deoxycholate, which facilitates the LOX-mediated peroxidation of the polyunsaturated fatty acids esterified to the PL. The LOX-peroxidized PI, after enzymatic catalysis, was separated from the unoxidized PI in the reaction mixture by normal-phase, high performance liquid chromatography (HPLC). The extent of LOX-mediated peroxidation of PI following HPLC purification was established by the analysis of lipid phosphorus, conjugated dienes by UV spectrophotometry, peroxides, and loss of fatty acids by gas chromatography. This study established the optimal conditions yielding approximately 46% of peroxidized PI from 300 microg of neat bovine liver PI that was peroxidized by soybean type-I LOX (50 microg) for 30 min in borate buffer (0.2 M, pH 9.0) containing 10 mM deoxycholate. PMID- 20013194 TI - Enzyme immunoassay of isoprostanes. AB - Isoprostanes are a group of prostaglandin-like compounds that are derived in vivo primarily by free radical-mediated peroxidation of arachidonic acid within phospholipids. The resulting isoprostane moieties are rapidly hydrolyzed, metabolized, and excreted. It is now well recognized that isoprostane levels, most commonly those of 15F(2t)-isoprostane (formerly denoted 8-iso-prostaglandin F(2) (alpha)) in blood or urine, are useful biomarkers for the assessment of oxidative stress, which is a major risk factor for many diseases. Isoprostanes were initially quantified by GC/MS following multiple extraction and purification steps. However, to facilitate routine cost-effective analysis of these biomarkers, immunoassays for isoprostanes were developed. Given the large number of isoprostanes, isoprostane metabolites, and isoprostane conjugates in vivo, the potential for their ex vivo generation from arachidonic acid, and differences among protocols for GC/MS and immunoassay of these biomarkers, it is not surprising that discrepancies arise in values obtained using different methods. When appropriate precautions are taken during sample collection, preparation, and analysis, enzyme immunoassays for 15F(2t)-isoprostane can be used for the reproducible and reliable assessment of oxidative stress in experimental animals or human subjects. PMID- 20013193 TI - Detection and localization of markers of oxidative stress by in situ methods: application in the study of Alzheimer disease. AB - Oxidative stress is a key factor involved in the development and progression of Alzheimer disease (AD), and it is well documented that free radical oxidative damage, particularly of neuronal lipids, proteins, nucleic acids, and sugars, is extensive in brains of AD patients. The complex chemistry of peroxynitrite has been the subject of intense study and is now evident that there are two principal pathways for protein modification: the first one involves homolytic hydroxyl radical-like chemistry that results in protein-based carbonyls and the second involves electrophilic nitration of vulnerable side chains, in particular the electron-rich aromatic rings of Tyr and Trp. In the presence of buffering bicarbonate, peroxynitrite forms a CO(2) adduct, which augments its reactivity. Formation of 3-nitrotyrosine by this route has become the classical protein marker specifically for the presence of peroxynitrite. Protein-based carbonyls can be detected by two methods: (i) derivatization with 2,4 dinitrophenylhydrazine (DNPH) and detection of the protein-bound hydrazones using an enzyme-linked anti-2,4-dinitrophenyl antibody and (ii) derivatization with biotin-hydrazide and detection of the protein-bound acyl hydrazone with enzyme linked avidin or streptavidin. Glycation of proteins by reducing sugars (Maillard reaction) results in a profile of time-dependent adduct evolution rendering susceptibility to oxidative elaboration. In addition, oxidative stress can result in oxidized sugar derivatives which can subsequently modify protein through a process known as glycoxidation. Of more general importance, oxidative stress results in lipid peroxidation and the production of a range of electrophilic and mostly bifunctional aldehydes that modify numerous proteins. The more important protein modifications are referred to as advanced glycation end products (AGEs) and advanced lipoxidation end products (ALEs). Protein modification can result in both non-cross-link and cross-link AGEs and ALEs, the latter arising from the potential bifunctional reactivity, such as that of the lipid-derived modifiers 4 hydroxy-2-nonenal (HNE) and malondialdehyde (MDA). Oxidative damage to nucleic acids results in base modification, substitutions, and deletions. Among the most common modifications, 8-hydroxyguanosine (8OHG) is considered a signature of oxidative damage to nucleic acid.Cells are not passive to increased oxygen radical production but rather upregulate protective responses. In neurodegenerative diseases, heme oxygenase-1 (HO-1) induction is coincident with the formation of neurofibrillary tangles. This enzyme that converts heme, a prooxidant, to biliverdin/bilirubin (antioxidants) and free iron has been considered an antioxidant enzyme. But seen in the context of arresting apoptosis, HO-1 and tau may play a role in maintaining the neurons free from the apoptotic signal (cytochrome c), since tau has strong iron-binding sites. Given the importance of iron as a catalyst for the generation of reactive oxygen species, changes in proteins associated with iron homeostasis can be used as an index of cellular responses. One such class of proteins is the iron regulatory proteins (IRPs) that respond to cellular iron concentrations by regulating the translation of proteins involved in iron uptake, storage, and utilization. Therefore, IRPs are considered to be the central control components of cellular iron concentration. PMID- 20013196 TI - Peptidomics: divide et impera. AB - The term "peptidomics" can be defined as the systematic analysis of the peptide content within a cell, organelle, tissue or organism. The science of peptidomics usually refers to the studies of naturally occurring peptides. Another meaning refers to the peptidomics approach to protein analysis. An ancient Roman strategy divide et impera (divide and conquer) reflects the essence of peptidomics. Most effort in this field is spent purifying and dividing the peptidomes, which consist of tens, hundreds or sometimes thousands of functional peptides, followed by their structural and functional characterisation. This chapter introduces the concept of peptidomics, outlines the range of methodologies employed and describes key targets - the peptide groups which are often sought after in such studies. PMID- 20013195 TI - Application of membrane extraction with sorbent interface for breath analysis. AB - The detection of volatile organic compounds (VOCs) in human breath may be useful for routine clinical diagnosis of several diseases in a non-invasive manner. Traditional methods of breath analysis have some technical limitations. Membrane extraction with a sorbent interface (MESI), however, has many advantages over current methods, including good selectivity and sensitivity, and is well suited for breath analysis.The aim of this project was to develop a simple and reproducible sampling device and method based on the MESI system for breath analysis. The feasibility and validity of the MESI system was tested with real human breath samples. The use of breath CO(2) as an internal standard for the analysis of breath VOCs is an effective method to solve the difficulties associated with variations in the target analyte concentrations in a sample, which are attributed to mass losses and different breathing patterns of different subjects. In this study, the concentration of breath acetone was successfully expressed normalized to CO(2) as in the alveolar air. Breath acetone of healthy males and females profiled at different times of the day was plotted using the MESI system, and results are consistent with literature. This technique can be used for monitoring breath acetone concentrations of diabetic patients and for applications with other biomarker monitoring. PMID- 20013192 TI - Oxidized low-density lipoprotein. AB - Oxidized low-density lipoprotein (Ox-LDL) has been studied for over 25 years. Numerous pro- and anti-atherogenic properties have been attributed to Ox-LDL. Yet, Ox-LDL has neither been defined nor characterized, as its components and composition change depending on its source, method of preparation, storage, and use. It contains unoxidized and oxidized fatty acid derivatives both in the ester and free forms, their decomposition products, cholesterol and its oxidized products, proteins with oxidized amino acids and cross-links, and polypeptides with varying extents of covalent modification with lipid oxidation products, and many others. It seems to exist in vivo in some form not yet fully characterized. Until its pathophysiological significance, and how it is generated in vivo are determined, the nature of its true identity will be only of classical interest. In this review, its components, their biological actions and methods of preparation will be discussed. PMID- 20013197 TI - Performing comparative peptidomics analyses of Salmonella from different growth conditions. AB - Host-pathogen interactions are complex competitions during which both the host and the pathogen adapt rapidly to each other in order for one or the other to survive. Salmonella enterica serovar Typhimurium is a pathogen with a broad host range that causes a typhoid fever-like disease in mice and severe food poisoning in humans. The murine typhoid fever is a systemic infection in which S. typhimurium evades part of the immune system by replicating inside macrophages and other cells. The transition from a foodborne contaminant to an intracellular pathogen must occur rapidly in multiple, ordered steps in order for S. typhimurium to thrive within its host environment. Using S. typhimurium isolated from rich culture conditions and from conditions that mimic the hostile intracellular environment of the host cell, a native low molecular weight protein fraction, or peptidome, was enriched from cell lysates by precipitation of intact proteins with organic solvents. The enriched peptidome was analyzed by both LC MS/MS and LC-MS-based methods, although several other methods are possible. Pre fractionation of peptides allowed identification of small proteins and protein degradation products that would normally be overlooked. Comparison of peptides present in lysates prepared from Salmonella grown under different conditions provided a unique insight into cellular degradation processes as well as identification of novel peptides encoded in the genome but not annotated. The overall approach is detailed here as applied to Salmonella and is adaptable to a broad range of biological systems. PMID- 20013198 TI - Approaches to identify endogenous peptides in the soil nematode Caenorhabditis elegans. AB - The transparent soil nematode Caenorhabditis elegans can be considered an important model organism due to its ease of cultivation, suitability for high throughput genetic screens, and extremely well-defined anatomy. C. elegans contains exactly 959 cells that are ordered in defined differentiated tissues. Although C. elegans only possesses 302 neurons, a large number of similarities among the neuropeptidergic signaling pathways can be observed with other metazoans. Neuropeptides are important messenger molecules that regulate a wide variety of physiological processes. These peptidergic signaling molecules can therefore be considered important drug targets or biomarkers. Neuropeptide signaling is in the nanomolar range, and biochemical elucidation of individual peptide sequences in the past without the genomic information was challenging. Since the rise of many genome-sequencing projects and the significant boost of mass spectrometry instrumentation, many hyphenated techniques can be used to explore the "peptidome" of individual species, organs, or even cell cultures. The peptidomic approach aims to identify endogenously present (neuro)peptides by using liquid chromatography and mass spectrometry in a high-throughput way. Here we outline the basic procedures for the maintenance of C. elegans nematodes and describe in detail the peptide extraction procedures. Two peptidomics strategies (off-line HPLC-MALDI-TOF MS and on-line 2D-nanoLC-Q-TOF MS/MS) and the necessary instrumentation are described. PMID- 20013199 TI - Mass spectrometric analysis of molluscan neuropeptides. AB - The central nervous systems of molluscan species contain high levels of structurally diverse peptides that function as neurotransmitters, neuromodulators or neurohormones. Peptide diversity is believed to be a way to increase the information handling capacity of neurons in the context of a brain with low cell numbers and neuronal connectivity. Accordingly, much effort has been made to identify peptides from single neurons and tissues of interest. In the past decade a mass spectrometry-based approach has been applied to detect and characterize peptides from single neurons, nerves and tissues of the molluscan brain. Peptides from single neurons are often analysed directly by mass spectrometry without prior sample preparation. Single neurons from the molluscan brain may be identified based on their position, cell morphology and colour. Neurons that cannot be readily identified can be tagged functionally or chemically. For the analysis of peptides from tissues, special extraction methods in conjunction with peptide separation by liquid chromatography coupled to mass spectrometry have been developed. Tens to hundreds of peptides from the tissue extract can be detected and characterized in a single analysis. PMID- 20013201 TI - Protocols for peptidomic analysis of spider venoms. AB - Spider venom contains a complex mixture of components with a large range of molecular masses (0.1-60 kDa) exhibiting a diverse array of actions. Most of these components are proteinaceous molecules - biologically active proteins and peptides. Proteomics profiling of spider venoms (the components with MW >10 kDa) could be achieved through conventional 2-DE-based proteomics methods combined with MS or MS/MS detection. Peptidomic profiling (of the components with MW below ~10 kDa) is usually achieved through off-line separation by a combination of ion exchange and reverse-phase chromatography, and it relies more heavily on de novo sequencing by Edman degradation or MS/MS for peptide identification. PMID- 20013202 TI - Purification and characterization of biologically active peptides from spider venoms. AB - Spider venoms represent invaluable sources of biologically active compounds suitable for use in life science research and also having a significant potential for biotechnology and therapeutic applications. The methods reported herewith are based on our long experience of spider venom fractionation and peptides purification. We routinely screen new peptides for antimicrobial and insecticidal activities and our detailed protocols are also reported here. So far these have been tested on species of Central Asian and European spiders from the families Agelenidae, Eresidae, Gnaphosidae, Lycosidae, Miturgidae, Oxyopidae, Philodromidae, Pisauridae, Segestriidae, Theridiidae, Thomisidae, and Zodariidae. The reported protocols should be easily adaptable for use with other arthropod species. PMID- 20013203 TI - MALDI-TOF mass spectrometry approaches to the characterisation of insect neuropeptides. AB - The diversity of insect neuropeptides coupled with the limitations from the small size of the insects themselves combine to make positive identification through peptide sequencing a highly challenging task. The advent of the "soft-ionisation" techniques of MALDI-TOF and electrospray (ESI)-Q-TOF mass spectrometry, coupled with the additional information from insect genome projects have revolutionised the characterisation of insect neuropeptides, such that sequences can now be obtained from just a few cells, where before thousands of insects had to be laboriously dissected, extracted and purified. Some of the procedures that are now used to identify these peptides are described here. Once the neuropeptides have been identified, it then becomes possible to use this knowledge to define physiological functionality. PMID- 20013200 TI - Monitoring neuropeptides in vivo via microdialysis and mass spectrometry. AB - Neuropeptides are important signaling molecules that regulate many essential physiological processes. Microdialysis offers a way to sample neuropeptides in vivo. When combined with liquid chromatography-mass spectrometry detection, many known and unknown neuropeptides can be identified from a live organism. This chapter describes sample preparation techniques and general strategies for the mass spectral analysis of neuropeptides collected via microdialysis sampling. Methods for the in vitro microdialysis of a neuropeptide standard as well as the in vivo microdialysis sampling of neuropeptides from a live crab are described. PMID- 20013205 TI - Direct peptide profiling of brain tissue by MALDI-TOF mass spectrometry. AB - Direct MALDI-TOF mass spectrometric peptide profiling is increasingly used to analyze the peptide complement in the nervous system of a variety of invertebrate animals from leech to Aplysia and many arthropod species, especially insects and crustaceans. Here, we describe a protocol for direct peptide profiling of defined areas of the central nervous system of insects. With this method, one can routinely and reliably obtain neuropeptide signatures of selected brain areas from various insects. PMID- 20013204 TI - Direct MALDI-TOF mass spectrometric peptide profiling of neuroendocrine tissue of Drosophila. AB - Direct MALDI-TOF mass spectrometric peptide profiling is increasingly used to analyze the peptide complement in the nervous system of a variety of invertebrate animals, from leech to Aplysia and many arthropod species, especially insects and crustaceans. Proper sample preparation is often the most crucial step to obtain the necessary data. Here, we describe protocols for the use of MALDI-TOF mass spectrometry to directly analyze the peptidome of neuroendocrine tissues of insects, particularly Drosophila melanogaster, by MALDI-TOF MS. PMID- 20013206 TI - Peptidomic analysis of single identified neurons. AB - Today, commercially available mass spectrometers increasingly meet all the demands of the proteomics community including high throughput, high sensitivity, and significant fragmentation capability for sequence determinations. Therefore, proper sample preparation is often the most crucial step to obtain the necessary data, particularly when working with biological samples. Depending on the size, sample preparation techniques differ and have to be optimized empirically. This is particularly apparent at the single cell level. In this chapter, we describe protocols for the use of MALDI-TOF mass spectrometry to directly analyse the peptidome of single insect neurons. PMID- 20013207 TI - Identification and analysis of bioactive peptides in amphibian skin secretions. AB - Skin secretions from anurans (frogs and toads), particularly those species belonging to the Hylidae and Ranidae families, are a rich source of biologically active peptides. Cytolytic peptides with broad-spectrum antimicrobial activities and highly variable amino acid sequences are often released into these secretions in high concentrations. Identification and characterization of these components can prove to be valuable in species identification, elucidation of evolutionary histories and phylogenetic relationships between species, and may lead to development of agents with potential for therapeutic application. This chapter describes the use of norepinephrine (injection or immersion) to stimulate peptide release in a procedure that does not appear to cause distress to the animals. The peptide components in the secretions are separated by reversed-phase HPLC on octadecylsilyl silica (C(18)) columns under standard conditions after partial purification on Sep-Pak cartridges. Individual peptides are identified by determination of their molecular masses by MALDI-TOF mass spectrometry and from their retention times. The use of mixtures of synthetic peptides of appropriate molecular mass as calibration standards enables mass determination to a high degree of precision. PMID- 20013208 TI - An efficient protocol for DNA amplification of multiple amphibian skin antimicrobial peptide cDNAs. AB - Antimicrobial peptides (AMPs) play an important role in the host's innate defence system in many organisms. Amphibian skin is expected to be a particularly rich source of novel AMPs. In amphibians, AMPs are produced from precursor proteins via specific cleavage by processing enzymes. While the nucleotide sequences of the AMP coding region in precursors are hypervariable, those of other regions, including the 5(')- and 3(')-untranslated regions (UTRs), are highly or relatively conserved in different precursors. Such nucleotide sequence conservation suggests an efficient strategy for molecular cloning of the antimicrobial peptide genes by 3(')-rapid amplification of cDNA ends (3(')-RACE) and reverse transcriptase polymerase chain reaction (RT-PCR) methods using specific primers. With this strategy in mind we have established an efficient protocol suitable for amplification of multiple cDNAs encoding amphibian AMP precursor proteins. PMID- 20013209 TI - Combined peptidomics and genomics approach to the isolation of amphibian antimicrobial peptides. AB - A large number of diverse antimicrobial peptides have been found in amphibian skins, and many more remain to be identified. It is sufficiently easy to obtain amounts of gland secretions sufficient for both identification and functional testing of the bioactive peptides. We describe here a systematic peptidomics approach which we combined with genomics and functional testing. This has proven to be an effective way to identify amphibian antimicrobial peptides, including novel peptide families. Protocols are exemplified for Bombina maxima and Odorrana grahami and can be easily adapted for use with other amphibian species. PMID- 20013210 TI - Identification and relative quantification of neuropeptides from the endocrine tissues. AB - Endocrine tissues like the pituitary, hypothalamus and islets of Langerhans are rich in bioactive peptides. These are used for intercellular signalling and are involved in regulation of almost all physiological processes. Peptidomics is the comprehensive analysis of peptides in tissues, fluids and cells. Peptidomics applied to (neuro-)endocrine tissues aims therefore to identify as many bioactive peptides as possible. Peptidomics of (neuro-)endocrine tissues requires an integrated approach that consists of careful sample handling, peptide separation techniques, mass spectrometry and bioinformatics. Here we describe the methods for isolation and dissection of endocrine tissues, the extraction of bioactive peptides and further sample handling and identification of peptides by mass spectrometry and hyphenated techniques. We also present a straightforward method for the comparison of relative levels of bioactive peptides in these endocrine tissues under varying physiological conditions. The latter helps to elucidate functions of the bioactive peptides. PMID- 20013211 TI - Peptidome analysis of mouse liver tissue by size exclusion chromatography prefractionation. AB - Here we report our approach to the peptidomic analysis of mouse liver. We use ultrafiltration for peptide prefractionation, which is followed by size exclusion chromatography. The low molecular weight peptides (MW below ~3 kDa) are analysed directly by nanoLC-MS/MS, and the higher molecular weight peptides (MW above ~3 kDa) are characterized with MALDI-TOF MS first and then proteolytically digested prior to nanoLC-MS/MS analyses. PMID- 20013212 TI - Rat brain neuropeptidomics: tissue collection, protease inhibition, neuropeptide extraction, and mass spectrometric analysis. AB - Due to the complexity of the mammalian central nervous system, neuropeptidomic studies in mammals often yield very complicated mass spectra that make data analysis difficult. Careful sample preparation and extraction protocols must be employed in order to minimize spectral complexity and enable extraction of useful information on neuropeptides from a given sample. Controlling post-mortem protease activity is essential to simplifying mass spectra and to identifying low abundance neuropeptides in tissue samples. Post-mortem microwave-irradiation coupled with cryostat dissection has proven to be effective in arresting protease activity to allow detection of endogenous neuropeptides instead of protein degradation products. PMID- 20013214 TI - Peptidomics analysis of lymphoblastoid cell lines. AB - A key challenge in clinics is the identification of sensitive and specific biomarkers for early detection, prognostic evaluation, and surveillance of disease. A biomarker is defined as a biological substance that can be used to specifically detect a disease, measure its progression, or the effect of a treatment. A biomarker should be easily accessible, and ideally sensitivity and specificity must be sufficient to distinguish between false positives, false negatives, and true positives. To be useful for routine clinical evaluation, a biomarker should be detectable in body fluids (e.g., plasma, serum, urine). A biomarker can be a metabolite, a specific post-translational modification, a lipid, a phospholipid, or a protein. Due to technical advances in the analysis of biomolecules by mass spectrometry (MS), investigations of peptide biomarkers have increased. In contrast to genome, the peptidome is dynamic and constantly changing. Elucidating how the peptides complement changes in a cell type in diseases is crucial to understand how these processes occur at a molecular level. Lymphoblastoid cell lines, derived from blood lymphocytes, represent suitable models for biochemical investigations and biomedical applications because of their stability, the ease of amplification, and long-term preservation. Technological improvements of MS and liquid chromatography (LC) during the last 10 years resulted in the development of highly sensitive approaches for proteomic and peptidomic analyses. Here we provide guidelines for the preparation of the lymphoblastoid cell lines, the extraction of the peptides and their purification. We describe a number of technologies which we developed for the peptidomic profiling of lymphoblastoid cell extracts from patients with leukodystrophies, linked to mutations in the genes encoding the eukaryotic initiation factor 2B (eIF2B; eIF2B-related disorders). PMID- 20013213 TI - Quantitative neuroproteomics of the synapse. AB - An emerging way to study neuropsychiatric or neurodegenerative diseases is by performing proteomic analyses of brain tissues. Here, we describe methods used to isolate and identify the proteins associated with a sample of interest, such as the synapse, as well as to compare the levels of proteins in the sample under different conditions. These techniques, involving subcellular fractionation and modern quantitative proteomics using isotopic labels, can be used to understand the organization of neuronal compartments and the regulation of synaptic function under various conditions. PMID- 20013215 TI - Peptidomics: identification of pathogenic and marker peptides. AB - Recent years have seen great advances in mass spectrometry and proteomics, the science dealing with the analysis of proteins, their structure and function. A branch of proteomics dealing with naturally occurring peptides is often referred to as peptidomics. Direct analysis of peptides produced by processing or degradation of proteins might be useful for example for detecting and identifying pathogenic and/or biomarker peptides in body fluids like blood. In this paper, we introduce one of the standard protocols for comprehensive analysis of serum derived peptides, which consists of methods for purification of serum peptides, detection of peptides, pattern recognition and clustering (bioinformatics), and identification of peptide sequences. Peptide identification should be followed by the investigation of their pathogenic roles using for example synthetic peptides and the establishment of their usefulness as bioclinical markers. PMID- 20013216 TI - Peptidomic approaches to the identification and characterization of functional peptides in Hydra. AB - Little is known about peptides that control developmental processes such as cell differentiation and pattern formation in metazoans. The cnidarian Hydra is one of the most basal metazoans and is a key model system for studying the peptides involved in these processes. We developed a novel peptidomic approach to the isolation and identification of functional signalling peptides from Hydra (the Hydra peptide project). First, peptides extracted from the tissue of Hydra magnipapillata are purified to homogeneity using high-performance liquid chromatography (HPLC). The isolated peptides are then tested for their ability to alter gene expression in Hydra using differential display-PCR (DD-PCR). If gene expression is altered, the peptide is considered as a putative signalling peptide and is subjected to amino acid sequencing. Following the sequencing, synthetic peptides are produced and compared to their native counterparts by HPLC and/or mass spectrometry (MS). The synthetic peptides, which are available in larger quantities than their native analogues, are then tested in a variety of biological assays in Hydra to determine their functions. Here we present our strategies and a systematic approach to the identification and characterization of novel signalling peptides in Hydra. We also describe our high-throughput reverse-phase nano-flow liquid chromatography matrix-assisted laser desorption ionization time-of-flight mass spectrometry (LC-MALDI-TOF-MS/MS) approach, which was proved to be a powerful tool in the discovery of novel signalling peptides. PMID- 20013217 TI - Immunochemical methods for the peptidomic analysis of tachykinin peptides and their precursors. AB - The tachykinins represent the largest known peptide family and are responsible for a range of pleiotropic functions in both vertebrates and invertebrates. Recent research has shown a diversity of mechanisms such as mRNA splicing, precursor processing and post-translation modification that can lead to a complex and continually expanding repertoire of tachykinin peptides. The peptidomic analysis of the tachykinins has been hindered by the lack of specific methodologies to capture, purify and characterise each tachykinin. This chapter summarises some of the methods that have been developed in order to further purify and characterise individual groups of tachykinin peptides from the peptidome. PMID- 20013218 TI - Affinity peptidomics: Peptide selection and affinity capture on hydrogels and microarrays. AB - Affinity peptidomics relies on the successfully proven approach used widely in mass-spectrometry-based protein analysis, where protein samples are proteolytically digested prior to the analysis. Unlike traditional proteomic analyses, affinity peptidomics employs affinity detection instead of, or in addition to, the mass-spectrometry detection. Affinity peptidomics, therefore, bridges the gap between protein microarrays and mass spectrometry and can be used for the detection, identification and quantification of endogenous or proteolytic peptides on microarrays and by MALDI-MS. Phage display technology is a widely applicable generic molecular display method suitable for studying protein-protein or protein-peptide interactions and the development of recombinant affinity reagents. Phage display complements the affinity peptidomics approach when the latter is used, e.g. to characterise a repertoire of antigenic determinants of polyclonal, monoclonal antibodies or other recombinantly obtained affinity reagents or in studying protein-protein interactions. 3D materials such as membrane-based porous substrates and acrylamide hydrogels provide convenient alternatives and are superior to many 2D surfaces in maintaining protein conformation and minimising non-specific interactions. Hydrogels have been found to be advantageous in performing antibody affinity assays and peptide-binding assays. Here we report a range of peptide selection and peptide-binding assays used for the detection, quantification or validation of peptide targets using array-based techniques and fluorescent or MS detection. PMID- 20013219 TI - In situ biosynthesis of peptide arrays. AB - Polypeptide and protein arrays enable high-throughput screening capabilities for studying molecular interactions and profiling of biomarkers, and provide a powerful functional screening tool for peptidomics. To overcome the limitations of conventional arraying methods, we have exploited cell-free systems for generating arrays of polypeptides by direct on-chip biosynthesis from DNA templates. Here we describe two protocols: (i) Protein In Situ Array (PISA), which allows the generation of polypeptide arrays in a single reaction by spotting cell-free lysate together with PCR DNA on a glass surface pre-coated with a capturing reagent, and (ii) DNA Array to Protein Array (DAPA), which is capable of producing multiple copies of a polypeptide array from a single DNA array template. The main advantage of these methods is in using an in vitro coupled transcription and translation system which circumvents the need to synthesise and purify individual polypeptides. Our methods allow making polypeptide arrays using amplified linear DNA fragments. PMID- 20013223 TI - Genetic mapping and positional cloning. AB - Genetic mapping and positional cloning of genetically complex traits in the laboratory rat (Rattus norvegicus) has recently led to the identification of various susceptibility genes in different rat models. Rat genetics has benefited from revolutionary advances in molecular biology, genetics, genomics and informatics and provide an unparalleled resource for molecular genetic investigation of mammalian physiopathology and its underlying complex genetic architecture. In this review, we will consider different strategies that are being used in the successful positional cloning of rat complex trait genes in the context of recent progress in rodent and human genetics. PMID- 20013222 TI - The rat: a model used in biomedical research. AB - The rat evokes fear and disgust in a large percentage of people around the world. Yet, other people are fascinated by this amazing creature that is raised as a pet, has an important place in several religions, and is a prominent model for biomedical research. This book focuses on a variety of methodologies that can be used in this remarkable model. This chapter sets the stage by providing a perspective on why the rat remains an important model in biomedical research. PMID- 20013221 TI - Bioinformatic identification of plant peptides. AB - Plant peptides play a number of important roles in defence, development and many other aspects of plant physiology. Identifying additional peptide sequences provides the starting point to investigate their function using molecular, genetic or biochemical techniques. Due to their small size, identifying peptide sequences may not succeed using the default bioinformatic approaches that work well for average-sized proteins. There are two general scenarios related to bioinformatic identification of peptides to be discussed in this paper. In the first scenario, one already has the sequence of a plant peptide and is trying to find more plant peptides with some sequence similarity to the starting peptide. To do this, the Basic Local Alignment Search Tool (BLAST) is employed, with the parameters adjusted to be more favourable for identifying potential peptide matches. A second scenario involves trying to identify plant peptides without using sequence similarity searches to known plant peptides. In this approach, features such as protein size and the presence of a cleavable amino-terminal signal peptide are used to screen annotated proteins. A variation of this method can be used to screen for unannotated peptides from genomic sequences. Bioinformatic resources related to Arabidopsis thaliana will be used to illustrate these approaches. PMID- 20013224 TI - Sequencing of the rat genome and databases. AB - The rat is an important system for modeling human disease. Four years ago, the rich 150-year history of rat research was transformed by the sequencing and annotation of the rat genome, ushering in an era of exceptional opportunity for identifying genes and pathways underlying disease phenotypes. With the genome sequence in place, there is the prospect of not only linking the extensive literature of mechanistic and pharmacological studies in the rat to its genome, but by using comparative genomics to other organisms as well. Genome-wide association studies (GWAS) in human populations have recently provided a direct approach for finding robust genetic associations in common diseases, but identifying the precise genes and their mechanisms of action remains challenging.The explosion of genomic tools and sequence over the last decade has created a wealth of data. Along with the data has arisen a need to manage it and to make it usable to scientists with a wide-range of research interests. This chapter is designed to overview the existing sequence and its utility, as well as provide a glimpse of some of the databases and bioinformatic tools available to the investigator. PMID- 20013220 TI - Bioinformatic approaches to the identification of novel neuropeptide precursors. AB - With the entire genome sequence of several animals now available, it is becoming possible to identify in silico all putative peptides and their precursors in an organism. In this chapter we describe a searching algorithm that can be used to scan the genome for predicted proteins with the structural hallmarks of (neuro)peptide precursors. We also describe how to use search strings such as the presence of a glycine residue as a putative amidation site, dibasic cleavage sites, the presence of a signal peptide, and specific peptide motifs to improve a standard BLAST search and make it suitable for searching (neuro)peptides in EST data. We briefly explain how bioinformatic tools and in silico predicted peptide precursor sequences can aid experimental peptide identification with mass spectrometry. PMID- 20013225 TI - Design of expression cassettes for the generation of transgenic animals (including insulators). AB - The use of transgenesis is relatively rare in rats, and this is because of the relative difficulty in adding foreign genes by the conventional methods. Gene knock out and knock in by the conventional techniques of homologous recombination remain difficult in rats. This situation would be less crucial if the gene constructs were more reliable for the expression of foreign genes. The present chapter describes the state of the art in vector design for various genetic modifications in rats. PMID- 20013226 TI - Inducible and conditional promoter systems to generate transgenic animals. AB - Transgenic animals are very useful models that can be utilized for the analysis of temporal and spatial gene expression in vivo. However, generation of a transgenic animal may become problematic if the presence of the transgene leads to conditions which are toxic or lethal to cell growth. In an effort to delineate the mechanism by which a specific gene contributes to cell growth and viability, an inducible and/or conditional system was established to generate transgenic animals. The systems comprise the following: (1) Selecting a specific promoter, (2) replacing a normal gene with other gene sequences (knock out), (3) promoting destruction of the mRNA (RNAi), (4) inducing and/or conditioning by drugs (Tet on/off), and (5) conditional cell knock out with cell death. The choice of system employed is dependent on the particular aim of the investigation, and may influence the final result. The inducible and conditional promoter system represents a useful experimental approach for the development of transgenic animals and the precise examination of gene function. PMID- 20013227 TI - Generation of transgenic rats by microinjection of short DNA fragments. AB - Here we describe an efficient technique to generate transgenic rats by microinjection of short DNA fragments. We have focused on optimal conditions for superovulation of prepubescent females Sprague-Dawley (CD) strains to have good quality embryos, pseudopregnant females, zygotes preparation, optimal conditions for microinjection and embryo transfer into foster mothers. PMID- 20013228 TI - Generation of transgenic rats using YAC and BAC DNA constructs. AB - Transgenic rats with a simple plasmid vector smaller than 20 Kb show insufficient expression and tissue specificity of the introduced transgene. Vectors derived from yeast artificial chromosome (YAC) and bacterial artificial chromosome (BAC), consisting of DNA fragments up to approximately 1 Mb (YAC) and approximately 200 Kb (BAC), respectively, and containing various endogenous regulatory sequences, were expected to work well and showed expression profiles comparable to their endogenous counterparts in transgenic animals. While attempting to make transgenic rats using YAC and BAC vectors, we faced two problems: how to prepare sufficiently concentrated intact DNA and how to reliably microinject a large DNA fragment into the fragile pronuclear ova of the rat. After solving these problems, we were able to make transgenic rats by introducing YAC/BAC gene constructs (YACs/BACs) into the pronuclear ova. And then we examined the relative transcription rates of these genes in the transgenic rats. In this chapter, we focus on this injection process. PMID- 20013232 TI - ENU mutagenesis to generate genetically modified rat models. AB - The rat is one of the most preferred model organisms in biomedical research and has been extremely useful for linking physiology and pathology to the genome. However, approaches to genetically modify specific genes in the rat germ line remain relatively scarce. To date, the most efficient approach for generating genetically modified rats has been the target-selected N-ethyl-N-nitrosourea (ENU) mutagenesis-based technology. Here, we describe the detailed protocols for ENU mutagenesis and mutant retrieval in the rat model organism. PMID- 20013230 TI - Generation of transgenic rats by ooplasmic injection of sperm cells exposed to exogenous DNA. AB - Intracytoplasmic sperm injection (ICSI) has been successfully achieved in mice and rats using a piezo-driven injection pipette, with the offspring rate of >30%. The ICSI technique was applied not only to rescue infertile male strains but also to produce transgenic rodents. The ICSI-mediated DNA transfer, that the sperm heads and exogenous DNA solution are mixed and co-injected into ooplasm, has been equally effective to the conventional pronuclear DNA microinjection. Production efficiency of transgenic founders by the ICSI-mediated DNA transfer was comparable between mice and rats, while the optimal DNA concentration was lower in rats than mice. The production efficiency was improved when membrane structure of sperm heads was partially disrupted by detergent or ultrasonic treatment before exposure to the exogenous DNA solution. Exogenous DNAs with various chain lengths were stably integrated into the rodent genomes of various genetic backgrounds by this method. The ICSI-mediated DNA transfer in which the preparation of pronuclear-stage fertilized zygotes is not required would be alternative to conventional pronuclear DNA microinjection. PMID- 20013229 TI - The use of lentiviral vectors to obtain transgenic rats. AB - Lentiviral vectors are now well recognized as good vehicles for gene delivery. This is because they can efficiently transduce both dividing and post-mitotic cells, and stably integrate into the host genome allowing for long-term expression of the transgene. Their potential utility for the generation of transgenic animals has been recognized as an attractive and promising alternative to the conventional DNA-microinjection method which lacks efficiency. The initial success of lentiviral transgenesis in mice considerably broadened its use in other species, in which classical transgenic techniques are difficult, such as in the rat.In this chapter, we describe detailed procedures for both the production of human immunodeficiency virus-1 (HIV-1)-derived lentiviral vectors and for the generation of transgenic rats by injection of these vectors into the perivitelline space of fertilized one-cell eggs. PMID- 20013231 TI - Procedures for somatic cell nuclear transfer in the rat. AB - Somatic cell nuclear transfer (SCNT) is a powerful tool for the investigation of the mechanisms of nuclear remodeling. In addition, SCNT may offer the possibility of introducing targeted mutations by homologous recombination in species for which ES cell technology is not available. The rat specific features of the oocyte have long impeded the development of SCNT. We detail here the procedures developed and optimized during the last several years for the optimization of rat cloning. PMID- 20013233 TI - Establishment of embryonic stem cells from rat blastocysts. AB - Rats have important advantages over mice as an experimental system for physiological and pharmacological investigations. Their embryonic stem (ES) cells, after differentiation into each tissue or organ, are applied in regenerative medicine, which enables examination of the effects of drugs for various diseases. Knockout rats will also provide a suitable model system for many human diseases and a great amount of new insights into gene functions, which have not been revealed by knockout mice. In 2008, we experienced the world's first success in establishing rat ES cells with chimeric contribution. Following on the heels of our report, others reported the establishment of rat ES cells that could complete a germline transmission. Recent studies on rat as well as mouse ES cells suggest that modifications of signal inhibitors and serum in the medium are critical for the maintenance of the pluripotency of ES cells. In this chapter, we discuss techniques for the successful establishment and maintenance of rat ES cells. PMID- 20013234 TI - Derivation, culture, and in vivo developmental capacity of embryonic cell lines from rat blastocysts. AB - Embryonic stem (ES) cells have been used extensively for site-specific gene targeting in the mouse. The resulting knock-out and knock-in mouse models generated so far have demonstrated their usefulness in biomedical research. However, for many diseases and fields of study, the rat still represents a superior model. The derivation and culture of germline-competent ES cells in the rat would allow the application of site-specific gene targeting technologies to this species of indisputable importance to biomedical research. We have recently shown the derivation, culture, and for the first time, in vivo contribution of rat ES-like cells to developing tissues. This represents an important step forward in making gene targeting technologies available to the rat research community, via development of rat ES cells. Here, we describe the materials, methods and techniques that have been used to obtain rat blastocysts, derive and culture embryonic cell lines from these, and assess the developmental capacity of the cells in vivo. PMID- 20013235 TI - Interference RNA for in vivo Knock-down of gene expression or genome-wide screening using shRNA. AB - With the lack of tools available to manipulate the rat genome, alternative technologies have been investigated to generate loss-of-function rat models by gene invalidation. The recent demonstration that RNA interference (RNAi)-mediated gene silencing occurs in rodents has opened new opportunities for rat functional genetics. In this chapter, we provide some practical guidelines for RNAi working in rat, based on the recent design and development of mice and rat Knock down models. PMID- 20013237 TI - Application of microarray-based analysis of gene expression in the field of toxicogenomics. AB - The field of toxicogenomics, which is becoming an important sub-discipline of toxicology, resulted from the natural convergence of the field of conventional toxicological research and the emergent field of functional genomics. One technology that has played a significant role in the field of toxicogenomics (in addition to many others) is the gene expression microarray. In this chapter, the authors provide an example of the application of gene expression microarrays to the field of toxicogenomics by detailing the strategy that was used for obtaining, analyzing, and interpreting gene expression data generated from RNA isolated from the liver of toxicant-exposed rats. PMID- 20013236 TI - Generation of gene-specific mutated rats using zinc-finger nucleases. AB - The genetic dissection of physiological and pathological traits in laboratory model organisms is accelerated by the ability to engineer loss-of-function mutations at investigator-specified loci. This chapter describes the use of zinc finger nucleases (ZFNs) for the targeted disruption of endogenous rat genes directly in the embryo. ZFNs can specifically disrupt target genes in cultured rat cells and in embryos from inbred and outbred strains, leading to permanently genetically modified animals. This technology allows for the rapid, targeted modification of the rat genome. PMID- 20013238 TI - Generation of congenic and consomic rat strains. AB - Congenic and consomic rat strains are inbred strains containing in their genome a given genomic region (congenic) or a whole chromosome (consomic) from another strain. They are nowadays invaluable tools for the identification of genes and mechanisms of multifactorial diseases, one of the main goals in biomedicine. They are produced by repeated backcrosses from a donor inbred strain to a recipient inbred strain, and thereafter maintained by conventional brother-x-sister mating. Although their production is lengthy and costly, it only requires a zootechny unit for breeding and tools for genotyping. PMID- 20013239 TI - Generation of rat "supersonic" congenic/conplastic strains using superovulation and embryo transfer. AB - Congenic strains are routinely used for positional mapping of quantitative trait loci; while conplastic strains, derived by substitution of different mitochondrial genomes on the same nuclear genetic background of inbred rodent strains, provide a way to unambiguously isolate effects of the mitochondrial genome on complex traits. Derivation of congenic or conplastic strains using a traditional backcross breeding strategy (10 backcrosses) takes more than 3 years. There are two principal strategies to speed up this process: (1) marker-assisted derivation of "speed" congenic/conplastic strains and (2) derivation of "supersonic" congenic/conplastic strains using in each backcross generation embryos obtained from 4-week-old superovulated females; thus, each backcross generation takes only 7 weeks. Both strategies could also be combined. In the current chapter, a method for derivation of "supersonic" congenic/conplastic rat strains is described. PMID- 20013240 TI - Analysis by quantitative PCR of zygosity in genetically modified organisms. AB - It is extremely useful to define a rapid and accurate method for identifying homozygous and heterozygous transgenic animals prior to setting up breeding programs for transgenic colonies and in experiments in which gene dosage effects could have a functional impact. Southern-blotting is a means of identifying zygosity, but such a method is time consuming and produces a high level of ambiguous results. Some years ago, we described the rapid, precise, non ambiguous, and high-throughput identification of zygosity in transgenic animals by real-time PCR. This technique allows us to make a clear-cut identification of transgenic rats, transgenic mice, and double-transgenic pigs. Since 2002, however, several authors have made improvements to this method. The following paper describes the ease with which zygosity is determined using real-time PCR. PMID- 20013241 TI - A restriction enzyme-PCR-based technique to determine transgene insertion sites. AB - Currently, most genetically engineered rat strains are created by methods that involve random integration of transgenes into the genome. The ability to identify the chromosomal location of the transgene insertion site enables the development of efficient genotyping assays, allows segregation of multiple transgene integration sites to be followed while breeding, and facilitates characterization of possible positional effects on phenotype. Here we describe a method for determining the chromosomal location of transgene insertion that combines restriction endonuclease enzyme digest with subsequent rounds of PCR amplification to produce amplicons representing the chromosomal regions flanking the integrated transgene. This method provides a reliable means for determining the exact location of insertion of transgenes within the genome. PMID- 20013242 TI - Cryopreservation and orthotopic transplantation of rat ovaries. AB - The number of rat strains increased considerably in the last decade and will increase continuously during the next years. This requires enough space for maintaining vital strains and techniques for cryobanking, which can be applied not only in specialised rat resource centres but also in regular animal houses. Here we describe an easy and fast method for the cryopreservation and transplantation of frozen-thawed ovaries of the rat. With dimethyl sulfoxide as cryoprotectant rat ovaries can be stored at -196 degrees C for unlimited time. For revitalisation thawed ovaries have to be orthotopically transplanted into appropriate ovarectomised recipients. Reestablishment of the reproductive cycle in the recipients can be confirmed by vaginal cytology shortly after transplantation. The recipients are able to produce 2-3 litters after mating with males of an appropriate strain. Cyropreservation of ovaries thus can be considered a reliable method to preserve scientifically and economically important stocks and strains of rats that are currently not required. PMID- 20013243 TI - Techniques for in vitro and in vivo fertilization in the rat. AB - Although in vitro and in vivo fertilization are powerful tools for restoring conserved sperm as well as stocked males in the rat, the techniques have progressively gained importance. However, the techniques are not used extensively for efficient production of rat offspring, because the techniques require a great deal of skill. This chapter describes the protocols for in vitro and in vivo fertilization in the rat. Namely, sperm collection, sperm cryopreservation, pre incubation of sperm, and insemination (co-culture with sperm and oocytes) for in vitro fertilization and intrauterine insemination for in vivo fertilization with fresh or frozen/thawed spermatozoa are provided. PMID- 20013244 TI - Rat strain repositories. AB - More than 500 inbred rat strains have been developed during the past 100 years for a wide range of biomedical applications. In addition to these traditionally bred strains, many induced mutants and several thousand mutagenized sperm samples have recently been generated. At present this huge number of strains is mainly managed by two rat resource centers, the National Bio Resource Project for the Rat in Japan (NBRP-Rat) and the US based Rat Resource and Research Center (RRRC). These resource centers not only collect, maintain and distribute rat strains as animals or cryopreserved embryos and spermatozoa, but also perform additional tasks such as phenotypic and genetic characterization as well as microbiological cleaning. Furthermore, they support researchers through informative databases in the selection of rat strains for specific research purposes. These global rat resource centers are essential for successful and sustainable research using the rat as a model species. PMID- 20013246 TI - Rat genomics applied to psychiatric research. AB - Psychiatric diseases are very debilitating and some of them highly prevalent (e.g., depression or anxiety). The rat remains one model of choice in this discipline to investigate the neural mechanisms underlying normal and pathological traits. Genomic tools are now applied to identify genes involved in psychiatric illnesses and also to provide new biomarkers for diagnostic and prognosis, new targets for treatment and more generally to better understand the functioning of the brain. In this report, we will review rat models, behavioral approaches used to model psychiatry-related traits and the major studies published in the field including genetic mapping of quantitative trait loci (QTL), transcriptomics, proteomics and transgenic models. PMID- 20013245 TI - Neurobehavioral tests in rat models of degenerative brain diseases. AB - Each translational approach in medical research forces the establishment of neurobehavioral screening systems, dedicated to fill the gap between postgenomic generation of state-of-the-art animal models (i.e. transgenic rats) on the one hand and their added value for really predictive experimental preclinical therapy on the other. Owing to these developments in the field, neuroscientists are frequently challenged by the task of detecting discrete behavioral differences in rats. Systematic, comprehensive phenotyping covers these needs and represents a central part of the process. In this chapter, we provide an overview on theoretical issues related to comprehensive neurobehavioral phenotyping of rats and propose specific classical procedures, protocols (similar to the SHIRPA approach in mice), as well as techniques for repeated, intraindividual phenotyping. Neurological testing of rats, motorfunctional screening using the accelerod approach, emotional screening using the social interaction test of anxiety, and testing of sensorimotoric gating functions by prepulse inhibition of the startle response are provided in more detail. This description is completed by an outlook on most recent developments in the field dealing with automated, intra-home-cage technologies, allowing continuous screening in rats in various behavioral and physiological dimensions on an ethological basis. PMID- 20013247 TI - Genomics studies of immune-mediated diseases using the BN-LEW rat model. AB - LEW and BN rats, that behave in opposite ways for their susceptibility to various immune-mediated diseases, provide a powerful model to investigate the molecular and genetic bases of immune system physiology and dysregulation. Using this model, we addressed the question of the genetic control of central nervous system autoimmunity, of xenobiotic-induced allergic diseases, and of T cell subsets that differ by their cytokine profiles. By linkage analysis and genetic dissection, using a panel of congenic rats, we identified a 120 Kb region on chromosome 9 that controls all these phenotypes, indicating that this region contains a gene or set of genes that plays an important role in the immune system homeostasis and susceptibility to immune mediated diseases. In this review, we will describe these rat genomics studies and will discuss the cellular and genetic factors that may be involved in the differences between these rat strains. PMID- 20013248 TI - Rat models of cardiovascular diseases. AB - In cardiovascular research, the rat has been the main model of choice for decades. Experimental procedures were developed to generate cardiovascular disease states in this species, such as systemic and pulmonary hypertension, cardiac hypertrophy and failure, myocardial infarction, and stroke. Furthermore, rats have been bred, which spontaneously develop such diseases. They became extremely valuable models to understand the genetics of these diseases, since powerful genomic tools are now available for the rat. One of these tools is transgenic technology, which has allowed the creation of even more disease models in the rat. This review summarizes the experimental, genetic, and transgenic rat models for cardiovascular diseases. PMID- 20013251 TI - Cancer research in rat models. AB - Rat has been the major model species used in several biomedical fields, notably in drug development and toxicology, including carcinogenicity testing. Rat is also a useful model in basic cancer research. Several rat models of monogenic (Mendelian) human hereditary cancers are available. Some were obtained spontaneously, while others were generated either by mutagenesis of tumor suppressor genes or by transgenesis of activated oncogenes (transgenesis can be performed efficiently in the rat). In addition, among the hundreds of inbred rat strains that have been isolated, some are highly susceptible or resistant to certain types of cancer, and these divergent phenotypes were shown to be polygenic. Numerous quantitative trait loci (QTLs) controlling cancer susceptibility/resistance have been defined in linkage analyses, and several of these QTLs were physically demonstrated in congenic strains. These studies led, in particular, to rapid translation to the human, with the identification of loci controlling susceptibility to a form of multiple endocrine neoplasia (monogenic trait) and to breast cancer (polygenic disease). The biology of cancer resistance has also been analyzed, and in some (but not all) cases, it was linked to regression of preneoplasic lesions. Rat tumors have been the subject of various types of analyses, and these studies led to important conclusions, including that tumors can be classified on the basis of the identity of the inducing agent, thereby suggesting that analyses of human tumors may be valuable in determining retrospectively the role of specific carcinogens in the formation of human cancers, and of human breast cancer in particular. PMID- 20013249 TI - Use of rat genomics for investigating the metabolic syndrome. AB - The spontaneously hypertensive rat (SHR) is the most widely used animal model of essential hypertension and accompanying metabolic disturbances. In this model, the use of whole genome sequencing and gene expression profiling techniques, linkage and correlation analyses in recombinant inbred strains, and in vitro and in vivo functional studies in congenic and transgenic lines has recently enabled molecular identification of quantitative trait loci (QTLs) relevant to the metabolic syndrome: (1) a deletion variant in Cd36 (fatty acid translocase) responsible for QTLs on chromosome 4 associated with dyslipidemia, insulin resistance and hypertension, (2) mutated Srebf1 (sterol regulatory element binding factor 1) as a QTL on chromosome 10 influencing dietary-induced changes in hepatic cholesterol levels, and (3) Ogn (osteoglycin) as a QTL on chromosome 17 associated with left ventricular hypertrophy. In addition, selective replacement of the mitochondrial genome of the SHR with the mitochondrial genome of the Brown Norway rat influenced several major metabolic risk factors for type 2 diabetes and provided evidence that spontaneous variation in the mitochondrial genome per se can promote systemic metabolic disturbances relevant to the pathogenesis of metabolic syndrome. Owing to recent progress in the development of rat genomic resources, the pace of QTL identification and discovery of new disease mechanisms can be expected to accelerate in the near future. PMID- 20013253 TI - [Hip dislocation and fracture of the femoral head caused by low impact trauma]. AB - Hip dislocations and particularly fractures of the femoral head are very rare injuries. Various therapy options are described. So far there has been consensus about the severity of the trauma which has causes this type of injury. This case study presents a type II Pipkin fracture. This specific "accident" was due to low impact trauma, which was previously considered to be insufficient to cause this type of injury. We report on the diagnostic and therapeutic measures and compare this procedure with information from the literature. PMID- 20013250 TI - Genomic research in rat models of kidney disease. AB - Current understanding of the mechanisms underlying renal disease in humans is incomplete. Consequently, our ability to prevent the occurrence of renal disease or treat kidney disease once it develops is limited. There are objective difficulties in investigating kidney disease directly in humans, leading investigators to resort to experimental animal models that simulate renal disease in humans. Animal models have thus been a tool of major importance in the study of normal renal physiology and have been crucial in shedding light on the complex mechanisms involved in normal kidney function and in our current understanding of and ability to treat renal disease. Among the animal models, rat has been the preferred and most commonly used species for the investigation of renal disease. This chapter reviews what has been achieved over the years, using rat as a tool for the investigation of renal disease in humans, focusing on the contribution of rat genetics and genomics to the elucidation of the mechanisms underlying the pathophysiology of the major types of renal disease, including primary and secondary renal diseases. PMID- 20013254 TI - [Neurogenetics of emotional processes. Neuroimaging findings as endophenotypes for depression]. AB - Major depression is one of the most frequent and serious psychiatric diseases. Although the disease is highly heritable, the search for candidate genes has been of limited success hitherto. The complex, polygenetic hereditary transmissions coding for heterogeneous, clinically defined phenotypes such as major depression may be better identified using the endophenotype approach. A recent study, reporting an association of the risk allele in a serotonin transporter polymorphism (5-HTTLPR) with increased amygdala responsiveness to aversive stimuli, stimulated the new research field of imaging genetics, which is characterized by the choice of neurobiological activity patterns as endophenotypes. This review discusses recent studies from this rapidly growing research field, focussing on genetic effects on cortico-limbic circuitries during emotion processing. Evidence is reviewed suggesting that potential risk-alleles for depression are associated with functional cortico-limbic abnormalities, which frequently occur in patients with major depression. PMID- 20013252 TI - [Giant congenital naevus : Indications and techniques for surgical treatment]. AB - According to current knowledge the term giant congenital naevus is generally applied to skin alterations which consist of naevus cells, that are already conspicuous at birth and reach a diameter of at least 20 cm or more in adulthood. Surgical removal of such alterations is fundamentally indicated because there is high potential for degeneration. The surgical challenge is the functional and aesthetic reconstruction after removal. This article presents the incidence, natural course and pathology of such giant congenital naevus alterations. The advantages and disadvantages of various reconstruction procedures are presented together with an algorithm for management of these potentially malignant alterations. PMID- 20013255 TI - The putative Bacillus subtilis L,D-transpeptidase YciB is a lipoprotein that localizes to the cell poles in a divisome-dependent manner. AB - Cell wall synthesis in bacteria is spatially organized by cytoskeletal structures. Common to all cell wall-bearing bacteria, the cytokinetic machinery localizes the cell wall synthesis to the site of septation. Recently, MinJ, a new component of the cytokinetic machinery, or divisome, of Bacillus subtilis has been described. MinJ is part of the division site selection system but also essential for correct assembly of the divisome. Here, I used the isolated PDZ domain of MinJ for co-elution experiments. One of the proteins that co-eluted was the so far uncharacterized, putative L,D-transpeptidase protein YciB. Evidence is shown that YciB localizes to the cell poles. YciB localization depends on the existence of a mature divisome, suggesting that L,D-transpeptidases are, like penicillin-binding proteins, part of the divisome. PMID- 20013256 TI - Bilateral thermal hyperalgesia in trigeminal and extra-trigeminal regions in patients with myofascial temporomandibular disorders. AB - Our aim was to assess thermal sensitivity in both trigeminal and extra-trigeminal regions in patients with myofascial temporomandibular disorder (TMD) but without comorbid conditions as compared to age-matched controls. Twenty women (age 24 +/- 3 years) diagnosed with myofascial TMD according to the research diagnostic criteria for TMD and 20 healthy women (age 24 +/- 4 years) were included. Warm and cold detection thresholds (WDT and CDT, respectively) and heat and cold pain thresholds (HPT and CPT, respectively) were bilaterally assessed over the masseter and frontalis muscles (trigeminal regions) and the wrist (extra trigeminal region). The mean of three determinations at each site was calculated and used for analysis. The order of the test sites was randomized. A two-way ANOVA was used to test for differences between groups and sides (most painful/contra-lateral side; dominant/non-dominant). No significant differences between groups for WDT or CDT in trigeminal and extra-trigeminal regions (ANOVA, P > 0.389) were found. There were significant differences between groups, but not between sides, for HPT and CPT in both trigeminal and extra-trigeminal areas (ANOVA, P < 0.001). CPT (P < 0.001) over the trigeminal area was positively correlated with both pain intensity and duration of pain symptoms: the longer the history of pain or the greater the pain intensity, the higher the CPT (i.e., the greater cold hyperalgesia) over the trigeminal region. Our findings revealed bilateral thermal hyperalgesia (lower HPT and higher CPT) but normal WDT and CDT in trigeminal and extra-trigeminal regions in women with myofascial TMD as compared to healthy controls. Bilateral heat/cold hyperalgesia in trigeminal and extra-trigeminal areas may reflect a dysfunction of thermal channels in myofascial TMD patients as result of some combination of peripheral sensitization, facilitation of central nociceptive processing and/or decreased descending inhibition. PMID- 20013259 TI - The case for postmortem imaging. PMID- 20013260 TI - A renaissance man. PMID- 20013258 TI - Current techniques in postmortem imaging with specific attention to paediatric applications. AB - In this review we discuss the decline of and current controversies regarding conventional autopsies and the use of postmortem radiology as an adjunct to and a possible alternative for the conventional autopsy. We will address the radiological techniques and applications for postmortem imaging in children. PMID- 20013261 TI - Arbuscular mycorrhizal fungi, Bacillus cereus, and Candida parapsilosis from a multicontaminated soil alleviate metal toxicity in plants. AB - We investigated if the limited development of Trifolium repens growing in a heavy metal (HM) multicontaminated soil was increased by selected native microorganisms, bacteria (Bacillus cereus (Bc)), yeast (Candida parapsilosis (Cp)), or arbuscular mycorrhizal fungi (AMF), used either as single or dual inoculants. These microbial inoculants were assayed to ascertain whether the selection of HM-tolerant microorganisms can benefit plant growth and nutrient uptake and depress HM acquisition. The inoculated microorganisms, particularly in dual associations, increased plant biomass by 148% (Bc), 162%, (Cp), and 204% (AMF), concomitantly producing the highest symbiotic (AMF colonisation and nodulation) rates. The lack of AMF colonisation and nodulation in plants growing in this natural, polluted soil was compensated by adapted microbial inoculants. The metal bioaccumulation abilities of the inoculated microorganisms and particularly the microbial effect on decreasing metal concentrations in shoot biomass seem to be involved in such effects. Regarding microbial HM tolerance, the activities of antioxidant enzymes known to play an important role in cell protection by alleviating cellular oxidative damage, such as superoxide dismutase, catalase, glutathione reductase, and ascorbate peroxidase, were here considered as an index of microbial metal tolerance. Enzymatic mechanisms slightly changed in the HM-adapted B. cereus or C. parapsilosis in the presence of metals. Antioxidants seem to be directly involved in the adaptative microbial response and survival in HM-polluted sites. Microbial inoculations showed a bioremediation potential and helped plants to develop in the multicontaminated soil. Thus, they could be used as a biotechnological tool to improve plant development in HM-contaminated environments. PMID- 20013257 TI - Pharmacokinetics of linezolid in septic patients with and without extended dialysis. AB - PURPOSE: Linezolid is an option for the treatment of infections caused by multiresistant Gram-positive bacteria. The survival of critically ill patients with acute renal failure (ARF) can be improved by increasing the dose of renal replacement therapy. Extended (daily) dialysis (ED) is a new and important approach to renal replacement therapy in intensive care units. The aim of the study was to evaluate the pharmacokinetics of linezolid in septic patients without ED and on ED, respectively. METHODS: We studied the pharmacokinetics of linezolid in adult intensive care patients with sepsis (n = 5) and anuric septic patients with ARF being treated with ED (n = 10). Linezolid 600 mg was administered intravenously twice daily. The pharmacokinetic parameters, their variability, and possible covariates were analyzed using NONMEM. RESULTS: The pharmacokinetics of linezolid followed a two-compartment model with clearance (Cl) = 0.159 L h(-1) kg(-1) +/- 51% (population mean +/- interindividual variability), central volume of distribution (V(1)) = 0.273 L/kg +/- 21%, intercompartmental clearance (Q) = 0.369 L h(-1) kg(-1), and peripheral volume of distribution (V(2)) = 0.271 L/kg. The clearance in ED patients while on dialysis was increased by 3.5 L/h, and patients with liver transplantation/resection had their clearance reduced by 60%. Intra-individual variability was much smaller than inter-individual variability. CONCLUSIONS: Our results suggest that linezolid pharmacokinetics in critically ill patients with ARF undergoing ED is not comparable to that in healthy subjects and patients without ARF. The best method of managing linezolid dosage in such a complex group of patients, whose physiology can vary daily, would be to use therapeutic drug monitoring. PMID- 20013265 TI - An unusual case of undifferentiated connective tissue disease presenting as cardiac tamponade. AB - Cardiac tamponade as an initial manifestation of undifferentiated connective tissue diseases (UCTD) is extremely rare, with only one case reported in literature thus far. We describe here, a case of a middle-aged man who presented with symptoms of fatigue, exertional dyspnea and orthopnea. His physical exam was significant for anasarca, elevated JVP and pulsus paradoxus. Chest X-ray showed pleural effusions and cardiomegaly, electrocardiogram revealed electrical alternans and a transthoracic echocardiogram demonstrated massive pericardial effusion with hemodynamic compromise. There was clear evidence of tamponade on right heart catheterization. All common causes of pericardial effusion were assiduously excluded before working up the patient for connective tissue disorders, which revealed a high antinuclear antibody titer (1:160), grossly elevated SSA, SSB antibodies and increased C-reactive protein levels (13.04 mg/dl). Patient had no signs or symptoms suggestive of systemic sclerosis (xerophthalmia or xerostomia) and did not meet criteria for any other known connective tissue diseases. He was therefore diagnosed with UCTD, and successfully treated with colchicine after emergency pericardiocentesis. This case presents UCTD as a rare cause of cardiac tamponade and large pericardial effusions and suggests that colchicine can be used to treat UCTD-associated effusions. These patients once diagnosed, are at risk of developing known connective tissue diseases within 5 years of disease onset and should be followed up in clinic periodically. PMID- 20013266 TI - Pentoxifylline for the treatment of anterior uveitis in Behcet's disease: possible alternative for TNF blockers. AB - Behcet's disease is an inflammatory disorder of unknown cause, characterized by recurrent oral aphthous ulcers, genital ulcers, uveitis, and skin lesions. All these common manifestations are self-limiting except for the ocular attacks. The objective of the present study is to report the efficacy of pentoxifylline for ocular involvement in Behcet's disease. We report a patient with ocular involvement secondary to Behcet's disease which was corticosteroid dependent and refractory to azathioprine treatment. Pentoxifylline was added with amelioration of inflammation. Pentoxifylline may be a useful treatment option in patients with Behcet's disease, where other immunosuppressives have failed or are contra indicated. PMID- 20013264 TI - Mimicry of lyme arthritis by synovial hemangioma. AB - To report on the differential diagnosis of lyme arthritis and synovial hemangioma due to similar clinical and radiological signs and symptoms. A 15-year-old boy presented at the age of 9 with recurrent rather painless swelling of the right knee. Altogether four episodes lasting for 1-2 weeks each occurred over a period of 18 months before medical advice was sought. Physical examination revealed only a slightly limited range of motion. Living in an endemic area of borreliosis, he reported a tick bite 6 months prior to onset of his symptoms with erythema migrans and was treated for 10 days with amoxicillin. Serology revealed two positive unspecific bands in IgG immunoblot (p41 and 66) with slight positivity for ELISA. Ultrasound revealed synovial thickening and increased fluid. Despite the weak positive serology a diagnosis of lyme arthritis could not be excluded and intravenous antibiotic treatment with ceftriaxone was started. After two further relapses antiinflammatory therapy including intraarticular steroids were introduced with no long lasting effect. A chronical disease developed with alternate periods of swelling and almost complete remission. Ultrasound as well as MRI demonstrated ongoing signs of synovitis, therefore after further progression, a diagnostic arthroscopy was performed showing an inconspicuous knee joint. A second MRI showed focal suprapatellar enhancement and was followed by open arthrotomy revealing a histopathological proven synovial cavernous juxtaarticular hemangioma. To our knowledge, the differential diagnosis of lyme arthritis and synovial hemangioma has not yet been reported despite obvious clinical similarities. In conclusion, in children and adolescents synovial hemangioma has to be considered in differential diagnosis of recurrent knee swelling. Early diagnosis is important to prevent prolonged suffering from chronic joint swelling with probable joint damages, unnecessary treatment procedures and as well school and sports absenteeism. PMID- 20013263 TI - [Solitary fibrous tumor and haemangiopericytoma: what is new?]. AB - Soft-tissue tumors with haemangiopericytoma (HPC)-like growth patterns can now be divided into three categories: (1) The solitary fibrous tumour (SFT) group with its variants; (2) lesions showing clear evidence of myoid/pericytic differentiation and corresponding to "true" HPCs (myopericytoma/glomangiopericytoma and a subset of sinonasal HPCs); (3) neoplasms that occasionally display HPC-like features (e.g. synovial sarcoma). In this study 268 intrathoracic and extrathoracic SFTs from the German consultation and reference center of soft tissue tumors in Jena were evaluated and analyzed immunohistochemically with antibodies CD34, Bcl-2, CD99, SMA, S100, PanCK and Ki 67. Furthermore, SFTs were categorized into the newly proposed SFT designation: Fibrous variant, cellular variant (more than 90% hypercellularity), fat-forming variant, giant cell-rich variant and malignant SFTs. This article should provide insights into the diagnosis of this entity with emphasis on the new international standard. PMID- 20013262 TI - Archaea, Bacteria, and algal plastids associated with the reef-building corals Siderastrea stellata and Mussismilia hispida from Buzios, South Atlantic Ocean, Brazil. AB - Reef-building corals may be seen as holobiont organisms, presenting diverse associated microbial communities. Best known is the symbiotic relationship with zooxanthellae, but Archaea, Bacteria, fungi, viruses, and algal plastids are also abundant. Until now, there is little information concerning microbial communities associated with Brazilian corals. The present study aims to describe the diversity of Archaea, Bacteria, and eukaryotic algal plastid communities associated with two sympatric species, Siderastrea stellata and Mussismilia hispida, from Southeastern Brazil, using 16S rRNA gene libraries. Since corals present a high number of other associated invertebrates, coral barcoding (COI) was performed to confirm the exclusive occurrence of coral DNA in our samples. Our analysis yielded 354 distinct microbial OTUs, represented mainly by novel phylotypes. Richness (Chao1 and ACE) and diversity (H') estimations of the microbial communities associated with both species were high and comparable to other studies. Rarefaction analyses showed that microbial diversity of S. stellata is higher than that of M. hispida. Libshuff comparative analyses showed that the highest microbial community similarity between the two coral species occurred in the bacterial libraries, while archaeal and plastidial communities were significantly different. Crenarchaeota dominated archaeal communities, while Proteobacteria was the most abundant bacterial phylum, dominated by alpha Proteobacteria. Plastids were also represented by novel phylotypes and did not match with any 16S rRNA sequences of Cyanobacteria and zooxanthellae from GenBank. Our data improves the pool of available information on Brazilian coral microbes and shows corals as sources of diverse prokaryotic and picoeukaryotic communities. PMID- 20013268 TI - Thoracic outlet syndrome due to an axillary band: what if it ruptures? PMID- 20013267 TI - Anti-TNF therapies: a comprehensive analysis of adverse effects associated with immunosuppression. AB - Knowledge and understanding about the immunosuppressive properties of anti-TNF therapies and the adverse effects these causes have advanced over the last 10 years since the first of these drugs was approved. These drugs work by inhibiting tumour necrosis factor (TNF) in the body, which plays an essential role in the immune response to invading pathogens. Anti-TNF drugs have therapeutic value because high levels of TNF are thought to be part of the pathophysiology of many chronic inflammatory disorders such as rheumatoid arthritis and Crohn's disease. Anti-TNF drugs are usually well-tolerated, however, there have been reports of many potentially serious adverse effects. This article will comprehensively analyse these adverse effects; the incidence, symptoms and mechanisms will be discussed. In addition, the contraindications of this class of drugs will be explored and the detection and prevention methods that should be put in place by health care professionals who treat patients on these drugs will be described. PMID- 20013269 TI - Microscopic polyangiitis in an adolescent presenting as severe anemia and syncope. AB - Microscopic polyangiitis (MPA) is an autoimmune systemic vasculitis of small vessels. The condition has been best characterized in older adults and little is known of the natural history of this disease in children and adolescents. In this report, a case of an adolescent presenting with symptomatic anemia and syncopal episodes is described. An extensive evaluation ultimately led to the diagnosis of MPA. The unique findings in this case and review of the literature are presented, outlining the variable clinical presentations and challenge of diagnosing this condition in pediatric patients. PMID- 20013270 TI - Non-Hodgkin's lymphoma following treatment with etanercept in ankylosing spondylitis. AB - Anti-TNF drugs may increase lymphoma risk in autoimmune rheumatic diseases, such as rheumatoid arthritis, but there have been no reports stating increased risk of lymphoma in ankylosing spondylitis (AS). Before 2 years, we had presented a case with AS developing Hodgkin's lymphoma following 6 months of etanercept treatment. Hereby, we present another case with AS developing non-Hodgkin's lymphoma (NHL), subsequent to 11 months of etanercept treatment. Pathological analysis revealed diffuse large B cell NHL. Although this is a report of a single case, cautious use of anti-TNF drugs is strongly recommended as they might cause lymphoma development even in AS. PMID- 20013274 TI - Evaluation of renal allograft function early after transplantation with diffusion weighted MR imaging. AB - AIMS: To determine the inter-patient variability of apparent diffusion coefficients (ADC) and concurrent micro-circulation contributions from diffusion weighted MR imaging (DW-MRI) in renal allografts early after transplantation, and to obtain initial information on whether these measures are altered in histologically proven acute allograft rejection (AR). METHODS: DW-MRI was performed in 15 renal allograft recipients 5-19 days after transplantation. Four patients presented with AR and one with acute tubular necrosis (ATN). Total ADC (ADC(T)) was determined, which includes diffusion and micro-circulation contributions. Furthermore, diffusion and micro-circulation contributions were separated, yielding the "perfusion fraction" (F(P)), and "perfusion-free" diffusion (ADC(D)). RESULTS: Diffusion parameters in the ten allografts with stable function early after transplantation demonstrated low variabilities. Values for ADC(T) and ADC(D) were (x10(-5) mm(2)/s) 228 +/- 14 and 203 +/- 9, respectively, in cortex and 226 +/- 16 and 199 +/- 9, respectively, in medulla. F(P) values were 18 +/- 5% in cortex and 19 +/- 5% in medulla. F(P) values were strongly reduced to less than 12% in cortex and medulla of renal transplants with AR and ATN. F(P) values correlated with creatinine clearance. CONCLUSION: DW-MRI allows reliable determination of diffusion and micro-circulation contributions in renal allografts shortly after transplantation; deviations in AR indicate potential clinical utility of this method to non-invasively monitor derangements in renal allografts. PMID- 20013271 TI - Elevated serum heme oxygenase-1 and insulin-like growth factor-1 levels in patients with Henoch-Schonlein purpura. AB - The aim of this study is to investigate the levels of heme oxygenase-1 (HO-1), insulin-like growth factor-1 (IGF-1) and oxidative stress parameters including malondialdehyde (MDA), total antioxidant capacity (T-AOC) and the activities of total superoxide dismutase (SOD), glutathione peroxidase (GSH-PX), catalase (CAT) for exploring the correlations between these biological indexes and the clinical manifestations in Henoch-Schonlein purpura (HSP) patients. 36 patients with different phases of HSP and 16 age- and sex-matched controls were enrolled. MDA level, T-AOC and the activities of SOD, GSH-PX and CAT were measured by spectrophotometry. HO-1 and IGF-1 levels were detected by enzyme-linked immunosorbent assay. Significant higher MDA level, lower T-AOC, SOD, GSH-PX activities were shown in active phase of HSP, respectively, compared with those in early resolution phase of HSP (p < 0.001, <0.001, 0.017, <0.001, respectively) and the control subjects (p < 0.001, <0.001, 0.01, 0.008, respectively). HO-1 (both p < 0.001) and IGF-1 (p < 0.001, 0.009, respectively) levels in active phase and early resolution phase of HSP were significantly higher than those in normal controls. The changes of HO-1 and IGF-1 levels were coincident with overall clinical scores (r = 0.71, p < 0.001; r = 0.615, p < 0.001, respectively). The HO-1 level was found as positive correlation with MDA levels (r = 0.395, p = 0.017), but negative correlations with T-AOC (r = -0.409, p = 0.013) and SOD activities (r = -0.352, p = 0.035). HO-1 and IGF-1 were possibly involved in the pathogenesis of HSP; they could be the marker for evaluating the severity of the disease. PMID- 20013273 TI - Comparison of 4 cm Z-axis and 16 cm Z-axis multidetector CT perfusion. AB - OBJECTIVE: The aim of the study was to compare 4 cm with 16 cm Z-axis coverage in the assessment of brain CT perfusion (CTP) using. 320 slice multidetector CT METHODS: A retrospective non-randomised review of CTP performed on MD320 CT between September 2008 and January 2009 was undertaken. Two experienced readers reviewed the studies along with the 4 cm and 16 cm Z-axis CTP image data set. The outcome parameters assessed were the extent of the original finding, any additional findings and a change of diagnosis. RESULTS: 14 out of 27 patients were found to have abnormal CTP (mean age 58.1 years, 9 male). The 16 cm Z-axis increased the accuracy of the infarct core in 78% and ischaemic penumbra quantification in 100% of the cases. It also diagnosed additional infarcts in the same vascular territory in 28% of cases and in a different vascular territory in 14%. CONCLUSIONS: The increased field of view with MD320 better defines the true extent of the infarct core and ischaemic penumbra. It also identified other areas of infarction that were not identified on the 4 cm Z-axis. PMID- 20013272 TI - Does joint alignment affect the T2 values of cartilage in patients with knee osteoarthritis? AB - OBJECTIVE: To assess the relationship between T2 values of femorotibial cartilage and knee alignment in patients with clinical symptoms of medial osteoarthritis (OA). METHODS: Twenty-four patients (mean age +/- standard deviation, 62.5 +/- 9.9 years) with clinical symptoms of medial knee OA, 12 with varus and 12 with valgus alignment of the femorotibial joint, were investigated on 3T MR using a 2D multi-echo spin echo (MESE) sequence for T2 mapping. Analysis of covariance, Spearman correlation coefficients, exact Mann-Whitney tests, and Fisher's exact tests were used for statistical analysis. RESULTS: Overall the T2 values of cartilage in the medial compartment (median +/- interquartile-range, 49.44 +/- 6.58) were significantly higher (P = 0.0043) than those in the lateral compartment (47.15 +/- 6.87). Patients with varus alignment (50.83 +/- 6.30 ms) had significantly higher T2 values of cartilage (P < 0.0001) than patients with valgus alignment (46.20 +/- 6.00 ms). No statistically significant association between the T2 values of cartilage (in either location) and the Kellgren Lawrence score was found in the varus or in the valgus group. CONCLUSION: T2 measurements were increased in medial knee OA patients with varus alignment, adding support to the theory of an association of OA and joint alignment. PMID- 20013277 TI - [Tapetoretinal dystrophy and unusual facial features in an 8-year-old boy]. AB - An 8-year-old boy showed multiple cardiovascular and urinogenital anomalies, neurological problems as well as endocrinological abnormalities even shortly after birth. The patient had ophthalmological peculiarities, such as eyelid cleft configuration, tapetoretinal dystrophy, retinal visual atrophy, early childhood converging strabismus with varying angle relationships, nystagmus, light sensitivity and substantial visual deficits. Finally a Kabuki syndrome could be confirmed by human genetic investigations (8q22-23 triplication). PMID- 20013276 TI - Image quality and diagnostic accuracy of unenhanced SSFP MR angiography compared with conventional contrast-enhanced MR angiography for the assessment of thoracic aortic diseases. AB - OBJECTIVES: The purpose of this study was to determine the image quality and diagnostic accuracy of three-dimensional (3D) unenhanced steady state free precession (SSFP) magnetic resonance angiography (MRA) for the evaluation of thoracic aortic diseases. METHODS: Fifty consecutive patients with known or suspected thoracic aortic disease underwent free-breathing ECG-gated unenhanced SSFP MRA with non-selective radiofrequency excitation and contrast-enhanced (CE) MRA of the thorax at 1.5 T. Two readers independently evaluated the two datasets for image quality in the aortic root, ascending aorta, aortic arch, descending aorta, and origins of supra-aortic arteries, and for abnormal findings. Signal-to noise ratio (SNR) and contrast-to-noise ratio (CNR) were determined for both datasets. Sensitivity, specificity, and diagnostic accuracy of unenhanced SSFP MRA for the diagnosis of aortic abnormalities were determined. RESULTS: Abnormal aortic findings, including aneurysm (n = 47), coarctation (n = 14), dissection (n = 12), aortic graft (n = 6), intramural hematoma (n = 11), mural thrombus in the aortic arch (n = 1), and penetrating aortic ulcer (n = 9), were confidently detected on both datasets. Sensitivity, specificity, and diagnostic accuracy of SSFP MRA for the detection of aortic disease were 100% with CE-MRA serving as a reference standard. Image quality of the aortic root was significantly higher on SSFP MRA (P < 0.001) with no significant difference for other aortic segments (P > 0.05). SNR and CNR values were higher for all segments on SSFP MRA (P < 0.01). CONCLUSION: Our results suggest that free-breathing navigator-gated 3D SSFP MRA with non-selective radiofrequency excitation is a promising technique that provides high image quality and diagnostic accuracy for the assessment of thoracic aortic disease without the need for intravenous contrast material. PMID- 20013275 TI - Comparison of left ventricular function assessment using phonocardiogram- and electrocardiogram-triggered 2D SSFP CINE MR imaging at 1.5 T and 3.0 T. AB - OBJECTIVE: As high-field cardiac MRI (CMR) becomes more widespread the propensity of ECG to interference from electromagnetic fields (EMF) and to magneto hydrodynamic (MHD) effects increases and with it the motivation for a CMR triggering alternative. This study explores the suitability of acoustic cardiac triggering (ACT) for left ventricular (LV) function assessment in healthy subjects (n = 14). METHODS: Quantitative analysis of 2D CINE steady-state free precession (SSFP) images was conducted to compare ACT's performance with vector ECG (VCG). Endocardial border sharpness (EBS) was examined paralleled by quantitative LV function assessment. RESULTS: Unlike VCG, ACT provided signal traces free of interference from EMF or MHD effects. In the case of correct R wave recognition, VCG-triggered 2D CINE SSFP was immune to cardiac motion effects even at 3.0 T. However, VCG-triggered 2D SSFP CINE imaging was prone to cardiac motion and EBS degradation if R-wave misregistration occurred. ACT-triggered acquisitions yielded LV parameters (end-diastolic volume (EDV), end-systolic volume (ESV), stroke volume (SV), ejection fraction (EF) and left ventricular mass (LVM)) comparable with those derived from VCG-triggered acquisitions (1.5 T: ESV(VCG) = (56 +/- 17) ml, EDV(VCG) = (151 +/- 32) ml, LVM(VCG) = (97 +/- 27) g, SV(VCG) = (94 +/- 19) ml, EF(VCG) = (63 +/- 5)% cf. ESV(ACT) = (56 +/- 18) ml, EDV(ACT) = (147 +/- 36) ml, LVM(ACT) = (102 +/- 29) g, SV(ACT) = (91 +/- 22) ml, EF(ACT) = (62 +/- 6)%; 3.0 T: ESV(VCG) = (55 +/- 21) ml, EDV(VCG) = (151 +/- 32) ml, LVM(VCG) = (101 +/- 27) g, SV(VCG) = (96 +/- 15) ml, EF(VCG) = (65 +/- 7)% cf. ESV(ACT) = (54 +/- 20) ml, EDV(ACT) = (146 +/- 35) ml, LVM(ACT) = (101 +/- 30) g, SV(ACT) = (92 +/- 17) ml, EF(ACT) = (64 +/- 6)%). CONCLUSIONS: ACT's intrinsic insensitivity to interference from electromagnetic fields renders it suitable for clinical CMR. PMID- 20013278 TI - Use of intrathecal urokinase in repeated shunt and external ventricular drain blockage from high CSF protein due to an optic pathway glioma. AB - INTRODUCTION: High cerebrospinal fluid (CSF) protein is a recognised association of optic pathway gliomas. This can occlude ventriculoperitoneal (VP) shunt catheters or external ventricular drains (EVD). CASE REPORT: We describe an 8 year-old boy with an optic pathway glioma, who had frequent episodes of a blocked VP shunt and EVD due to high CSF protein level. This responded favourably to repeated urokinase instillation into the catheter lumen. We present the course of treatment and the method of administration and review the literature behind thrombolytic therapy for occluded catheter devices. CONCLUSION: Intrathecal urokinase was effective in unblocking the EVD and lysing the protein clots within the ventricle. We found no previously published cases of intrathecal urokinase for this problem. This novel intervention seemed promising as a safe and effective means of maintaining EVD patency in cases complicated by excessive protein loads in CSF and hydrocephalus. PMID- 20013279 TI - What should we do with a discontinued shunt? AB - PURPOSE: The reported rate is up to 10% of shunt disconnection or fracture, either ventriculoperitoneal or subduroperitoneal. However, not all of shunt discontinuity is associated with shunt malfunction. We analyzed the discontinuity of the shunt system and related factors and tried to present a follow-up policy. METHODS: This is a retrospective study involving 22 patients who presented with shunt disconnection because of malfunction or other reasons between January 2003 and October 2008. To evaluate shunt function, we performed a shuntogram, temporary ligation, or intraoperative exploration. RESULTS: Nine (40.9%) of 22 patients had nonfunctioning shunts. The shunt system was removed in eight cases, and one patient refused surgical management. Of these 22 patients with disconnected shunts, 17 shunts placed in the occipital area were discontinued. The discontinuations were mostly fractured at neck (52%), not disconnected at connection site. CONCLUSION: Shunt disconnection has not been always represented a shunt malfunction. We thought many patients with disconnected shunt may be independent from shunt system, and it can be a good chance for patient to remove the shunt system. PMID- 20013281 TI - Intracardiac migration of the distal catheter of ventriculoperitoneal shunt: a case report. AB - INTRODUCTION: Migration of the distal catheter into the heart is an extremely rare complication of ventriculoperitoneal shunt, with only 11 cases reported in literature; in only four of them, there was also the migration into the pulmonary artery. The authors report the case of a young patient with this rare complication that was managed simply converting the ventriculoperitoneal shunt into a ventriculoatrial shunt. DISCUSSION: The migration of the distal catheter occurred through the internal jugular vein probably as the consequence of direct damage of the vessel during tunneling. Venous flow and negative intrathoracic pressure may have gradually pulled the catheter up into the heart; the upward migration may also be encouraged by the continuous movements of the patient that was agitated in the postoperative period as the consequence of her psychiatric disorder. The authors review the literature regarding migration of distal tubing into the heart and pulmonary artery. Diagnosis and management of this complication is discussed. PMID- 20013280 TI - Evaluation of nonnociceptive sensation in patients with congenital insensitivity to pain with anhidrosis. AB - PURPOSE: Congenital insensitivity to pain with anhidrosis (CIPA) is a rare, autosomal recessive disorder characterized by lack of pain and thermal sensation, anhidrosis, thermodysregulation, and mental retardation. Although nonnociceptive sensation, which is mediated by large-caliber myelinated Abeta fibers, is reported to be normal in CIPA patients, precise clinical assessments of this type of sensation have yet to be performed. The aim of this study was to evaluate peripheral sensory nerve function, including senses of touch/pressure, vibration, joint position, and two-point discrimination, in patients with CIPA by basic clinical neurological examination. METHODS: We examined touch/pressure sense, deep senses (vibration, joint proprioception), and two-point discrimination in 12 patients with CIPA (six males and six females aged 11-44) and 12 age- and sex matched healthy controls. Touch/pressure sense was examined with Semmes-Weinstein monofilaments, vibration sense with a tuning fork, and two-point discrimination with a vernier caliper. Joint proprioception was assessed through subject recognition of passive movement of the great toe. RESULTS: Perception thresholds of touch/pressure, vibration, and two-point discrimination were significantly higher (p < 0.05), and proprioception sensitivity was significantly lower (p < 0.05) in CIPA patients than in the healthy controls. CONCLUSIONS: Our findings suggest that CIPA patients suffer from more widespread disturbances of sensation than has been previously recognized. Impairment may not be restricted to the types of sensation conducted by peripheral sensory Adelta and C fibers. PMID- 20013282 TI - The incidence of Chiari malformation in nonsyndromic, single suture craniosynostosis. AB - INTRODUCTION: This study was designed to determine the incidence of Chiari malformation (CM) in nonsyndromic single suture craniosynostosis (N-SSSC). MATERIALS AND METHODS: A retrospective analysis of brain magnetic resonance imaging (MRI) studies of children undergoing craniofacial surgery during 1 January, 2004-31 March, 2009 in Cleft Palate and Craniofacial Centre, Department of Plastic Surgery, Helsinki University Hospital, Helsinki, Finland, was conducted. RESULTS AND DISCUSSION: One hundred twenty-four N-SSSC patients were imaged using brain MRI. Of these 124 patients, seven patients were diagnosed with N-SSSC with an associated CM; the incidence CM in association with N-SSSC was thus 5.6%. The average age at the time of MRI was 37.7 months. All were males, except one. Only two types of synostosis were identified in this patient cohort: synostosis involving the sagittal suture in five cases and right coronal synostosis in two cases. The CM malformations were relatively large. The tonsillar herniation varied from 6 to 12 mm (median 9 mm). All these patients were asymptomatic of their CMs. None had operation designed directly to correct the CM. CONCLUSION: As a conclusion, we state that the association of CM and N SSSC is a relatively common finding, with an incidence of 5.6% in preoperative brain MRI. The significance of finding CM in preoperative brain MRI should be assessed individually in asymptomatic patients. PMID- 20013283 TI - Intracranial hydatid cyst in children: report of 30 cases. AB - PURPOSE: To analyze the clinical manifestations, radiological features, and surgical outcomes in 30 pediatric intracranial hydatid cysts. METHODS: We reviewed the clinical, radiological, and surgical aspects of pediatric intracranial hydatid cysts patients who received surgical treatment at the Neurosurgical department of Xinjiang Medical University between the years 1985 to 2007, retrospectively. RESULTS: Headache and vomiting were the most common initial symptoms in our series. Neurological deficits from the mass effect of the cysts were seen in 15 cases, including hemiparesis, visual deficit, and diplopia. Epilepsy occurred only in one patient with temporal lobe hydatid cyst. On computed tomography (CT), it presented as a round-shaped and thin-walled homogeneous low-density cystic lesion without surrounding edema and enhancement. Only five patients had a magnetic resonance imaging (MRI) scan, and presented low signal intensity on T1-weighted image and high signal intensity on T2-weighted image. Surgical removal of cyst was performed in all cases and intact removal was done in 29 cases. However, one cyst ruptured during the dissection of cyst wall, thus, resulting in one death. There were no additional neurological deficits which were caused directly by surgery. CONCLUSION: Increased intracranial pressure is common in patients with cerebral hydatid disease. CT and MRI are the first-line diagnostic procedures. Surgery is the treatment of choice for the majority of intracranial hydatid cysts. Multiple and deep seated lesions should receive medical treatment postoperatively. PMID- 20013284 TI - [Critical overview of outcome parameters for patients with primary Sjogren's syndrome]. AB - Not only in the context of clinical trials in particular, but also in daily clinical practice, outcome parameters or measuring instruments are essential to assess the efficacy of a therapeutic intervention, its influence on disease activity and potentially also to predict further disease course. Such criteria can assist in the identification of patient risk groups that may require special checkups or interventions. Moreover, these parameters should be reliable, objective and valid, e.g. to allow comparison of results from different studies. Therefore, outcome parameters need to be developed and/or validated in a targeted manner for individual diseases or investigations. To date, we have only limited therapeutic options for Sjogren's syndrome, a frequent systemic autoimmune disorder of unknown origin. Against the background of the new therapy approaches expected, this article provides a critical overview of available and newly developed outcome parameters for patients with Sjogren's syndrome. PMID- 20013285 TI - [Current therapeutic options in Sjogren's syndrome]. AB - Sjogren's syndrome is a systemic inflammatory-rheumatic disorder of hitherto unknown origin and, hence, adequate therapy options are not available in most cases. Treatment of sicca symptoms in Sjogren's patients is primarily symptomatic. Glucocorticoids, NSAIDs and/or immunosuppressive drugs may be used for the treatment of extraglandular manifestations or complications. Although there have been few clinical studies to date, new insights into the pathogenesis of this disorder may permit novel therapeutic strategies. Targeting B-cell candidates as a new therapeutic option in this entity has shown promising results. B-cell-depletion using Rituximab has been tested in initial clinical trials for the treatment of Sjogren patients, both with and without associated B cell non-Hodgkin-lymphoma; however, further studies as well as reliable outcome criteria are needed. Further therapeutic options are currently in development or early clinical testing. PMID- 20013286 TI - Involvement of peptidylarginine deiminase-mediated post-translational citrullination in pathogenesis of sporadic Creutzfeldt-Jakob disease. AB - Peptidylarginine deiminases (PADs)-mediated post-translational citrullination processes play key roles in protein functions and structural stability through the conversion of arginine to citrulline in the presence of excessive calcium concentrations. In brain, PAD2 is abundantly expressed and can be involved in citrullination in disease. Recently, we have reported pathological characterization of PAD2 and citrullinated proteins in scrapie-infected mice, but the implication of protein citrullination in the pathophysiology in human prion disease is not clear. In the present study, we explored the molecular and biological involvement of PAD2 and the pathogenesis of citrullinated proteins in frontal cortex of patients with sporadic Creutzfeldt-Jakob disease (sCJD). We found increased expression of PAD2 in reactive astrocytes that also contained increased levels of citrullinated proteins. In addition, PAD activity was significantly elevated in patients with sCJD compared to controls. From two dimensional gel electrophoresis and MALDI-TOF mass analysis, we found various citrullinated candidates, including cytoskeletal and energy metabolism-associated proteins such as vimentin, glial fibrillary acidic protein, enolase, and phosphoglycerate kinase. Based on these findings, our investigations suggest that PAD2 activation and aberrant citrullinated proteins could play a role in pathogenesis and have value as a marker for the postmortem classification of neurodegenerative diseases. PMID- 20013287 TI - The association between educational level and age at the menopause: a systematic review. AB - OBJECTIVES: To evaluate the association between educational level and age at menopause, and to describe the potential mechanisms involved. METHODS: A systematic review of the literature was undertaken to evaluate this question. Educational level, which is considered one of the best socioeconomic indicators, was evaluated in this review as an exposure factor leading menopause to occur at an earlier age. The electronic search strategy was developed specifically for the different databases considered and through cross-referencing. After initially identifying 795 papers, 56 of them were selected for evaluation of the complete text. RESULTS: In the end, our systematic review included 25 papers covering 29 studies. In 75.9% of the studies evaluated in this review, there was a weak association (statistically significant in 45%) between lower educational level and earlier age of the menopause. The various methodological differences between the studies were unable to substantially influence the results. The way in which educational level was measured and the methodology for comparing strata differed greatly. The complexity of socioeconomic level mechanisms on menopause imposed limitations on the conclusions from this study. CONCLUSIONS: No unequivocal evidence for the existence of an association between educational level and age at menopause was found through this review. PMID- 20013288 TI - The epidemiology of abnormal hemoglobins in Mediterranean high-level athletes. AB - The aim of this study was to determine the prevalence and nature of hemoglobin (Hb) defects in a Mediterranean high-level (HL) athlete population. Five hundred and ninety-four HL male and female athletes were recruited during the annual follow-up of the members of Tunisian national teams. Hematological data, Hb electrophoresis, and DNA analysis were assessed using conventional techniques. Sporting discipline, type of sport, and performance levels were assessed using a questionnaire. The results showed that 32 HL athletes had abnormal Hb (5.4%): beta-thalassemia (2.2%), alpha-thalassemia (0.5%), HbAS (1.5%), HbAC (0.5%), and rare Hb variants (0.7%). Of the 32 defect carriers, all but one (a alpha thalassemia) were heterozygous. All the detected hemoglobinopathies but one (an Hb Hope) had already been reported in the country. The prevalence of Hb defect in the HL athletes was similar to that described in the general Tunisian population (P > 0.05). The percentage of Hb defect in the athletes was not dependent on gender, or performance level (P > 0.05). Within each type of sport the percentages of athletes with normal and abnormal Hb were similar (P > 0.05). The hematological data revealed the diversity of anemia, microcytosis, and hypochromia in thalassemic HL athletes. We concluded that HL athletes in Tunisia were a representative sample of the general Tunisian population regarding the prevalence and nature of benign abnormal Hb. The hematological data of the thalassemia carriers exhibited high variability and raised the question of genetic and sporting counseling, as well as biological follow-up for these carriers. PMID- 20013289 TI - Increased serum adenosine and interleukin 10 levels as new laboratory markers of increased intra-abdominal pressure. AB - BACKGROUND: Increased intra-abdominal pressure (IAP), intra-abdominal hypertension (IAH) and abdominal compartment syndrome (ACS) are severe complications of surgical interventions with a high rate of mortality. The technique of IAP measurement is accurate, precise, reproducible and cost effective. However, laboratory measures for monitoring of IAH have not been defined. We investigated the linkage between the serum levels of adenosine and interleukin 10 (IL-10) with IAP. METHODS: The sera of 25 surgical patients with IAP <12 mmHg and of 45 surgical patients with IAP >12 mmHg were tested. Serum adenosine concentration was measured by HPLC. Serum IL-1beta, IL-2, IL-4, IL-10, TNFalpha, IFNgamma and IL-10 were determined by enzyme linked immunosorbent assay (ELISA). CRP was measured by nephelometry. RESULTS: Significant correlations of IAP were found only with serum levels of adenosine and IL-10. In the sera of patients with IAP >12 mmHg, the levels of both adenosine (1.61 versus 0.06 uM, p < 0.01) and IL-10 (63.23 versus 27.27 pg/ml, p < 0.01) were significantly higher than those in patients with IAP <12 mmHg. Moreover, significant correlations were found between individual patient IAP-adenosine values (r = 0.766, p < 0.001), IAP IL-10 values (r = 0.792, p < 0.001) and adenosine-IL-10 values (r = 0.888, p < 0.001). A direct linear correlation between IAP-adenosine and IAP-10 values was only observed with IAP >15 (Grade II-IV). CONCLUSION: We report associations between IAP and the serum adenosine and IL-10 levels providing new tools for the laboratory monitoring of IAH as well as further understanding of the pathomechanisms contributing to ACS. PMID- 20013290 TI - Widespread and multifocal carcinomas in situ (CISs) through almost the entire pancreas: report of a case with preoperative cytological diagnosis. AB - PURPOSE: It is imperative for prognostic improvement of pancreatic cancer that we try to diagnose carcinoma in situ (CIS) of lesions, i.e., precursors of invasive ductal carcinomas (IDCs) at an early stage, because results of treatment of patients with IDCs themselves continue to be rather unsatisfactory. MATERIALS AND RESULTS: We report here a case of a patient who received subtotal pancreatectomy for widespread and multifocal CISs of the pancreas after preoperative brushing cytology from the epithelium of dilated main pancreatic duct proved cancer positive preoperatively. CONCLUSIONS: From our experience, we conclude that examination for CIS of the pancreas must be recommended whenever dilatation of relatively large pancreatic ducts is found by ultrasound or computed tomography. We should therefore advance to magnetic resonance cholangiopancreatography or endoscopic retrograde cholangiopancreatography and then cytological and/or pathological assessment of the pancreatic duct whenever non-continuous narrowing, localized dilatation, or other irregularities are encountered. PMID- 20013291 TI - Variability of surgical knot tying techniques: do we need to standardize? AB - INTRODUCTION: Knot tying is regarded as a basic surgical skill. However, there is no consensus in the literature on the best/safest technique. The aim of this study was to survey the various techniques used by German surgeons at the different stages of their training. MATERIALS AND METHODS: A knot tying technique was defined based on three criteria, i.e., both thread ends are crossed at the beginning at least two throws tied pulling at the same free end of the thread (standing part) and at least one knot tied changing the standing part. The knot tying techniques of 192 randomly chosen German surgeons practicing in 102 different hospitals were compared against this standard. Besides the status of the participating surgeons, the type of hospital in which they were currently working was recorded. RESULTS: Of the 192 participants, 69 were consultants, 56 fellows, and 67 residents. Forty-three surgeons (22.4%) worked in primary care centers (PCC), 81 (42.2%) in secondary care centers (SCC), and 68 (35.4%) in tertiary care centers (TCC). 62.5% crossed both thread ends at the beginning, 75.5% performed at least two identical throws keeping the same standing part, and 45.3% changed the standing part. Assessed against the technique defined as standard, 19.8% of all surgeons performed similar surgical knots. Even though consultants crossed both thread ends at the beginning and changed the standing part significantly more often (p = 0.007 and p = 0.031 respectively, Fisher's exact test), the overall rate of identical knots was independent of the status of participants and the type of hospital (p > 0.1, Fisher's exact test). CONCLUSION: This explorative study detected a high variability of knot tying techniques among German surgeons. In the absence of any evidence of the superiority of one particular technique, continuing education should focus on the quality of each single knot and perfect the details of the technique used. PMID- 20013292 TI - 8th International Symposium on Growth and Nutrition in Children with Chronic Renal Disease: 28-30 May 2009, Oviedo, Asturias, Spain. PMID- 20013293 TI - What have 20 years of data from the North American Pediatric Renal Transplant Cooperative Study taught us about growth following renal transplantation in infants, children, and adolescents with end-stage renal disease? AB - Growth following renal transplantation in infants, children, and adolescents was evaluated from 20 years of data reported to the registry of the North American Pediatric Renal Transplant Cooperative Study (NAPRTCS). The analysis of more than 10,000 recipients addressed the following questions: 1. What is the impact of age, pubertal growth, gender, transplant history, donor source and allograft function on growth after transplantation? 2. Has the height Z score at the time of transplantation changed during the past two decades and has this influenced final adult height? 3. To what extent has recombinant human growth hormone (rhGH) been utilized in growth retarded recipients after transplantation and has its use resulted in accelerated post-transplantation growth? 4. Has the use of steroids for maintenance immunosuppression changed over the past 20 years and how have the perturbations of steroid usage influenced post-transplantation growth? 5. Have changes in clinical care resulted in improved final adult height Z score during the past two decades? Only younger children (<6 years) had initial accelerated post-transplantation growth. The mean increment in height during puberty was 18.8 cm (21.7 cm in 4.7 years for boys and 14.3 cm in 4.5 years for girls). Gender, source of donor graft, or number of grafts did not influence growth. Height Z score at transplantation has improved over the past two decades, as has final adult height with each succeeding era. The use of rhGH after transplantation results in a delta Z score of +0.5 standard deviation (SD). Post-transplantation growth improves with steroid avoidance and changes in estimated glomerular filtration rate (eGFR) impact on growth. PMID- 20013294 TI - Reduced renal length and volume 20 years after very preterm birth. AB - Intrauterine growth retardation is presumed to be associated with decreased renal size and impaired renal function as a result of stunted kidney development and nephron deficit. To study whether very preterm birth also affects renal size at young adulthood, we sonographically measured bipolar kidney length and volume in 51 very premature individuals (<32 weeks of gestation), either small (SGA) or appropriate (AGA) for gestational age (22 SGA and 29 AGA), and 30 full-term controls 20 years after birth. Relative kidney length and volume were calculated. Both absolute and relative left kidney length and volume were significantly lower in SGA and AGA individuals, notably in women. Renal size did not differ between SGA and AGA individuals. In 70% of controls, the left kidney was larger than the right one compared with 40.9% in SGA [relative risk (RR) 1.7; 95% confidence interval (CI) 1.0-3.0] and 48.3% in AGA (RR 1.5; 95% CI 0.9-2.3) individuals. Renal structural anomalies were present in eight prematurely born participants only. Our data suggest that kidney growth is stunted after preterm birth, especially on the left side, and in the female gender. PMID- 20013295 TI - The appearance of rosette-like esophageal folds ("esophageal rosette") in the lower esophagus after a deep inspiration is a characteristic endoscopic finding of primary achalasia. AB - BACKGROUND: In healthy subjects who inspire deeply the lower esophagus usually opens, and the esophageal palisade vessels (EPVs) become visible. However, in patients with achalasia, the full extent of the EPVs does not become visible and, in addition, rosette-like esophageal folds appear in the lower esophagus. The aim of this study was to investigate whether or not these changes at the lower esophagus are characteristic findings of achalasia. METHODS: A total of 34 patients with achalasia and no esophageal dilatation following deep inspiration were compared with 34 sex- and age-matched control subjects. Following a deep inspiration, the lower esophagus of all study cohorts was evaluated on (1) whether or not the full extent of the EPVs was visible, (2) whether or not rosette-like esophageal folds appeared in the lower esophagus, and (3) whether or not there were any gastric lesions. RESULTS: One patient had secondary achalasia, and the remaining 33 patients had primary achalasia. In the control subjects, the full extent of the EPVs was clearly visible after a deep inspiration, and no esophageal folds appeared in the lower esophagus. In contrast, in the achalasia patients, EPVs were not observed in all patients after a deep inspiration, and rosette-like esophageal folds appeared in 33 of the 34 patients. CONCLUSION: After a deep inspiration, the non-visibility of the EPVs and the appearance of rosette-like esophageal folds at the lower esophagus, which we have called "esophageal rosette", are characteristic endoscopic findings of primary achalasia. PMID- 20013296 TI - Multiple extensor tendon ruptures caused by tophaceous gout. AB - A 54-year-old woman presented with painless subcutaneous masses on her right hand and spontaneous loss of extension of her ring finger. Surgical exploration of the wrist showed hypertrophic white-colored crystal deposits that both surrounded and invaded into the extensor tendons (intratendinous invasion). Histopathologic examination of the specimen demonstrated findings consistent with gouty tophi. We conclude that tophaceous gout needs to be included in the differential diagnosis of chronic extensor tenosynovitis of the hand. PMID- 20013297 TI - Seasonal changes in food resource distribution and feeding sites selected by Japanese macaques on Koshima Islet, Japan. AB - Feeding sites of Japanese macaques (Macaca fuscata) change according to seasonal fluctuations in food resource distribution. To examine what characteristics of food items affect feeding site selection, I describe herein the seasonal changes in food items, feeding sites, and food resource distributions of this species. Feeding behavior of monkeys and their food resource distributions were investigated on Koshima Islet, southern Japan, for four study periods (i.e., seasons) in 2002. Monkeys showed large variations in their diet between seasons. To weigh the relative influence of the distribution and abundance of food items on feeding site selection in each season, multiple regression analyses were performed by 100 m x 100 m grid. In the analyses, feeding time was a dependent variable and the abundance of staple food items, for which feeding time was over 5% in each season, in each grid square was an independent variable. There was no correlation between the resource distribution of most food items and the distribution of feeding time by monkeys in each season. Monkeys spent more feeding time where multiple staple food items were available. Food items that affected feeding site selection by monkeys had the following three characteristics: (1) clumped distribution, (2) seasonal availability, and (3) fruit. This suggests that monkeys are likely to select feeding sites to consume food items whose availability is limited temporally and spatially, which may enable them to simultaneously use other widely distributed, abundant food items efficiently. PMID- 20013299 TI - Enhanced tolerance of transgenic tall fescue plants overexpressing 2-Cys peroxiredoxin against methyl viologen and heat stresses. AB - Plant 2-Cys peroxiredoxins (2-Cys Prx) has both peroxidase and chaperon function. We overexpressed an Arabidopsis 2-Cys Prx in transgenic tall fescue (Festuca arundinacea) plants to confer tolerance against heat and methyl viologen (MV) stress. Transgenic plants were generated by Agrobacterium-mediated genetic transformation, and integration and expression of the transgene was confirmed by Southern, northern and western blot analyses. Compared to control plants, transgenic plants had significantly less electrolyte leakage and thiobarbituric acid-reactive substances (TBARS) when exposed to heat or MV. Under heat stress (42 degrees C), transgenic plants maintained their chlorophyll fluorescence (Fv/Fm) for 24 h while control plants lost chlorophyll fluorescence very quickly. We conclude that the high levels of 2-Cys Prx proteins in transgenic plants protect leaves from oxidative damage probably due to chaperon activity. PMID- 20013298 TI - Occipital gamma activation during Vipassana meditation. AB - Long-term Vipassana meditators sat in meditation vs. a control rest (mind wandering) state for 21 min in a counterbalanced design with spontaneous EEG recorded. Meditation state dynamics were measured with spectral decomposition of the last 6 min of the eyes-closed silent meditation compared to control state. Meditation was associated with a decrease in frontal delta (1-4 Hz) power, especially pronounced in those participants not reporting drowsiness during meditation. Relative increase in frontal theta (4-8 Hz) power was observed during meditation, as well as significantly increased parieto-occipital gamma (35-45 Hz) power, but no other state effects were found for the theta (4-8 Hz), alpha (8-12 Hz), or beta (12-25 Hz) bands. Alpha power was sensitive to condition order, and more experienced meditators exhibited no tendency toward enhanced alpha during meditation relative to the control task. All participants tended to exhibit decreased alpha in association with reported drowsiness. Cross-experimental session occipital gamma power was the greatest in meditators with a daily practice of 10+ years, and the meditation-related gamma power increase was similarly the strongest in such advanced practitioners. The findings suggest that long-term Vipassana meditation contributes to increased occipital gamma power related to long-term meditational expertise and enhanced sensory awareness. PMID- 20013300 TI - Effect of bacterial cell size on electricity generation in a single-compartmented microbial fuel cell. AB - A single-compartmented microbial fuel cell composed of a graphite felt anode modified with Neutral Red (NR-anode) and a porous Fe(II)-carbon cathode (FeC cathode) were compared for electricity generation from Microbacterium sp. and Pseudomonas sp. under identical conditions. Pseudomonas sp. was more than four times the size of Microbacterium sp. based on SEM images. In cyclic voltammetry, the redox reaction between Microbacterium sp and electrode was three times the rate observed between Pseudomonas sp. and the electrode based on the Y-axis (current) variation of cyclic voltammogram. The electric power generated by Microbacterium sp. was approx 3-4 times higher than that with Pseudomonas sp. during incubation for more than 150 days in the fuel cell. PMID- 20013301 TI - Green energy from Rhodopseudomonas palustris grown at low to high irradiance values, under fed-batch operational conditions. AB - Rhodopseudomonas palustris was grown under continuous irradiances of 36, 56, 75, 151, 320, 500, and 803 W m(-2), for a co-production of both bio-H(2) and biodiesel (lipids) using fed-batch conditions. The highest overall bio-H(2) produced [4.2 l(H(2)) l(culture) (-1)] was achieved at 320 W m(-2), while the highest dry biomass (3.18 g l(-1)) was attained at 500 W m(-2). Dry biomass contained between 22 and 39% lipid. The total energy conversion efficiency was at its highest (6.9%) at 36 W m(-2). PMID- 20013302 TI - Enhanced expression of heterologous inulinase in Kluyveromyces lactis by disruption of hap1 gene. AB - Inulinase gene (Kcinu) derived from Kluyveromyces cicerisporus was expressed extracellularly in Kluyveromyces lactis using an episomal vector directed by Kcinu promoter. The influence of hap1 gene disruption on the expression of inulinase was studied. Inulinase activity in the supernatant of the recombinant Klhap1Delta strain was 391 U ml(-1) after cultured 120 h, which was 2.2-fold that of the wild type host. The relative inulinase mRNA level of the Klhap1Delta strain was 11.3-fold that of the wild type strain, and the expression plasmid was more stable in the mutant host. Based on these results, the disruption of hap1 facilitated the high and stable expression of inulinase controlled by Kcinu promoter in K. lactis. PMID- 20013303 TI - Bio-resolution of glycidyl (o, m, p)-methylphenyl ethers by Bacillus megaterium. AB - A newly isolated Bacillus megaterium with epoxide hydrolase activity resolved racemic glycidyl (o, m, p)-methylphenyl ethers to give enantiopure epoxides in 84 99% enantiomeric excess and with 21-73 enantiomeric ratios. The (S)-enantiomer was obtained from rac-glycidyl (o or m)-methylphenyl ether while the (R)-epoxides was obtained from glycidyl p-methylphenyl ether. The observations are explained at the level by enzyme-substrate docking studies. PMID- 20013304 TI - Predictive performance for population models using stochastic differential equations applied on data from an oral glucose tolerance test. AB - Several articles have investigated stochastic differential equations (SDEs) in PK/PD models, but few have quantitatively investigated the benefits to predictive performance of models based on real data. Estimation of first phase insulin secretion which reflects beta-cell function using models of the OGTT is a difficult problem in need of further investigation. The present work aimed at investigating the power of SDEs to predict the first phase insulin secretion (AIR (0-8)) in the IVGTT based on parameters obtained from the minimal model of the OGTT, published by Breda et al. (Diabetes 50(1):150-158, 2001). In total 174 subjects underwent both an OGTT and a tolbutamide modified IVGTT. Estimation of parameters in the oral minimal model (OMM) was performed using the FOCE-method in NONMEM VI on insulin and C-peptide measurements. The suggested SDE models were based on a continuous AR(1) process, i.e. the Ornstein-Uhlenbeck process, and the extended Kalman filter was implemented in order to estimate the parameters of the models. Inclusion of the Ornstein-Uhlenbeck (OU) process caused improved description of the variation in the data as measured by the autocorrelation function (ACF) of one-step prediction errors. A main result was that application of SDE models improved the correlation between the individual first phase indexes obtained from OGTT and AIR (0-8) (r = 0.36 to r = 0.49 and r = 0.32 to r = 0.47 with C-peptide and insulin measurements, respectively). In addition to the increased correlation also the properties of the indexes obtained using the SDE models more correctly assessed the properties of the first phase indexes obtained from the IVGTT. In general it is concluded that the presented SDE approach not only caused autocorrelation of errors to decrease but also improved estimation of clinical measures obtained from the glucose tolerance tests. Since, the estimation time of extended models was not heavily increased compared to basic models, the applied method is concluded to have high relevance not only in theory but also in practice. PMID- 20013305 TI - The role of Ile476 in the structural stability and substrate binding of human cytochrome P450 2C8. AB - The biological function and stability of a cytochrome P450 (CYP) mainly depend on the subtle properties of the residues in the active site cavity, which are generally more divergent among proteins than other parts of the protein. As the most unique member of human CYP2C family, CYP2C8 has an isoleucine (Ile) 476 instead of phenylalanine (Phe) in substrate recognizing site 6 (SRS6). However, the role of Ile476 of CYP2C8 is still unknown. Therefore, six site-directed mutants of CYP2C8 were constructed to better define this. By UV-visible and circular dichroism spectroscopy studies, we studied for the first time the structural stability and all-trans-retinoic acid binding capability of the CYP2C8 variants. We found that the ferric CYP2C8 went through three states during thermal unfolding. Combined with substrate binding studies, our data revealed that residue 476 was involved in contact with substrate and was important for maintaining the thermal stability of CYP2C8. PMID- 20013306 TI - Are extended twin family designs worth the trouble? A comparison of the bias, precision, and accuracy of parameters estimated in four twin family models. AB - The classical twin design (CTD) uses observed covariances from monozygotic and dizygotic twin pairs to infer the relative magnitudes of genetic and environmental causes of phenotypic variation. Despite its wide use, it is well known that the CTD can produce biased estimates if its stringent assumptions are not met. By modeling observed covariances of twins' relatives in addition to twins themselves, extended twin family designs (ETFDs) require less stringent assumptions, can estimate many more parameters of interest, and should produce less biased estimates than the CTD. However, ETFDs are more complicated to use and interpret, and by attempting to estimate a large number of parameters, the precision of parameter estimates may suffer. This paper is a formal investigation into a simple question: Is it worthwhile to use more complex models such as ETFDs in behavioral genetics? In particular, we compare the bias, precision, and accuracy of estimates from the CTD and three increasingly complex ETFDs. We find the CTD does a decent job of estimating broad sense heritability, but CTD estimates of shared environmental effects and the relative importance of additive versus non-additive genetic variance can be biased, sometimes wildly so. Increasingly complex ETFDs, on the other hand, are more accurate and less sensitive to assumptions than simpler models. We conclude that researchers interested in characterizing the environment or the makeup of genetic variation should use ETFDs when possible. PMID- 20013307 TI - An online preconcentration system for the determination of uranium in water and effluent samples. AB - Amberlite XAD-4 resin functionalized with beta-nitroso-alpha-naphthol was applied to an online system for the preconcentration and determination of uranium. U (VI) ions were retained on the minicolumn at an appropriate pH and then desorbed with acid solution. The amount of uranium in the eluate was measured spectrophotometrically at 650 nm using Arsenazo III as a colorimetric reagent. The limit of detection and the preconcentration factor were 1.8 MUg L(-1) and 10, respectively. The chemical and flow variables affecting the preconcentration were studied. The influence of several ions on the system was also investigated. The method was successfully applied for the evaluation of uranium in water and in effluent samples. PMID- 20013308 TI - Variable selection in the accelerated failure time model via the bridge method. AB - In high throughput genomic studies, an important goal is to identify a small number of genomic markers that are associated with development and progression of diseases. A representative example is microarray prognostic studies, where the goal is to identify genes whose expressions are associated with disease free or overall survival. Because of the high dimensionality of gene expression data, standard survival analysis techniques cannot be directly applied. In addition, among the thousands of genes surveyed, only a subset are disease-associated. Gene selection is needed along with estimation. In this article, we model the relationship between gene expressions and survival using the accelerated failure time (AFT) models. We use the bridge penalization for regularized estimation and gene selection. An efficient iterative computational algorithm is proposed. Tuning parameters are selected using V-fold cross validation. We use a resampling method to evaluate the prediction performance of bridge estimator and the relative stability of identified genes. We show that the proposed bridge estimator is selection consistent under appropriate conditions. Analysis of two lymphoma prognostic studies suggests that the bridge estimator can identify a small number of genes and can have better prediction performance than the Lasso. PMID- 20013309 TI - Protection of mice against Brucella abortus 544 challenge by vaccination with recombinant OMP28 adjuvanted with CpG oligonucleotides. AB - Brucella abortus, a gram negative, facultative intracellular pathogen causes brucellosis in many animal species and humans. Although live, attenuated vaccines are available against this infection, they suffer from certain limitations. Therefore, the development of an effective subunit vaccine against brucellosis is an area of intense research. The outer membrane proteins (OMPs) of Brucella species have been extensively studied for its immunogenicity and protective ability. We have investigated the potential of CpG ODN to enhance the immunogenicity and protective efficacy of recombinant 28 kDa outer membrane protein (rOMP28) of Brucella melitensis. The study demonstrated vigorous immunoglobulin G (IgG) response of OMP28. The administration of rOMP28 with CpG caused increased cell mediated immune response in terms of induced IgG2a, T-cell proliferation and up-regulation of type I cytokine expression. In contrast, the free antigen suppressed the interferon gamma (type I cytokine) production on in vitro stimulation of spleenocytes. The result indicates the role of OMP28 in the down regulation of IFN-gamma production. Moreover, the B. abortus S-19 vaccinated mice showed highest production of IL-4 and IFN-gamma. The protective ability of the antigen was evaluated by systemic bacterial clearance after challenging the mouse with B. abortus 544 pathogen. The level of protection was significant in rOMP28+CpG treated mice but was lower than the required level. The results of the present study indicate that rOMP28 could be an immunogen capable of inducing both humoral and cellular immune response. The humoral response was biased towards Th1 type when it was co-administered with CpG. PMID- 20013311 TI - Wireless capsule endoscopy in enteropathy induced by nonsteroidal anti inflammatory drugs in pigs. AB - AIM: The aim of this study is to evaluate the diagnostic yield of capsule endoscopy in nonsteroidal anti-inflammatory drug (NSAID)-induced enteropathy in pigs. MATERIALS AND METHODS: Indomethacin (400 mg/day) was administrated orally for 10 days to eight female pigs weighing 36.3+/-2.4 kg. Afterwards, capsule endoscopy was performed, using the EndoCapsule system (Olympus Optical Co., Tokyo, Japan). The following morning, pharmacological euthanasia and immediate autopsy were performed. RESULTS: Small bowel injury compatible with NSAID-induced enteropathy was observed in 7/8 animals. The most common lesions were red spots and erosions. Ulcers and small intestinal bleeding were identified sporadically. Sensitivity and specificity of capsule endoscopy were 83.3% and 95.8%, respectively. CONCLUSION: Our results indicate that wireless capsule endoscopy is a highly accurate noninvasive method for evaluation of experimental NSAID-induced enteropathy. PMID- 20013310 TI - Eimeria bovis meront I-carrying host cells express parasite-specific antigens on their surface membrane. AB - Host immune responses conducted against antigens of Eimeria bovis are key factors for the development of protective immunity against this protozoan disease. In this study we investigated the expression of E. bovis-derived antigens on the host cell surface membrane during E. bovis first merogony in vitro. Host cells carrying E. bovis-meront I stages expressed E. bovis host cell surface antigens (EbHCSAg) on their surface membrane which were recognised by hyperimmune sera of calves and by sera from rats immunized with E. bovis merozoites I, when tested by indirect immune fluorescent antibody test (IIFAT), laser scanning confocal microscopy (LSCM) and immune electron microscopy. Expression of EbHCSAg on permissive host cells was earliest detected 7 days p. i., thus coinciding with the onset of the parasite replication. Membrane-associated EbHCSAg were removed from infected host cells by proteinase K, partially by Triton X-100, Triton X-114 and Triton X-405, but not by 1 M NaCl, CHAPS or phospholipase C treatment. Antibodies, affinity-purified on paraformaldehyde/glutardialdehyde (PAGA)-fixed E. bovis meront I-infected bovine host cells bound to the surface meront I carrying cells and to merozoites I (IIFAT, LSCM) but, in contrast to untreated sera, not to sporozoites. When tested on methanol-fixed merozoites I and sporozoites by IIFAT, affinity-purified antibodies bound to structures in the apical complex area of merozoites I, but not to sporozoites, whilst untreated sera caused diffuse labelling of internal structures of both parasite stages. Immune electron microscopy demonstrated binding of affinity-purified antibodies to micronemes and dense granules of merozoites I. Although the function of EbHCSAg is still unknown, results of this study might suggest an involvement in the development of protective immunity against E. bovis infections. PMID- 20013313 TI - A microfluidic device for depositing and addressing two cell populations with intercellular population communication capability. AB - We present a method for depositing cells in the microchambers of a sealed microfluidic device and establishing flow across the chambers independently and serially. The device comprises a transparent poly(dimethylsiloxane) (PDMS) microfluidic network (MFN) having 2 cell chambers with a volume of 0.49 microL, 6 microchannels for servicing the chambers, and 1 microchannel linking both chambers. The MFN is sealed with a Si chip having 6 vias and ports that can be left open or connected to high-precision pumps. Liquids are drawn through each chamber in parallel or sequentially at flow rates from 0.1 to 10 microL min(-1). Plugs of liquid as small as 0.5 microL can be passed in one chamber within 5 s to 5 min. Plugs of liquid can also be introduced into a chamber for residence times of up to 30 min. By injecting different liquids into 3 ports, 3 adjacent laminar streams of liquid can be drawn inside one chamber with lateral concentration gradients between the streams ranging from 20 to 500 microm. The flexibility of this device for depositing cells and exposing them to liquids in parallel or serially is illustrated by depositing two types of cells, murine N9 microglia and human SH-S5Y5 neuroblastoma. Microfluidic communication between the chambers is illustrated by stimulating N9 microglia using ATP to induce these cells to release plasma membrane vesicles. The vesicles are drawn through the second chamber containing neuroblastoma and collected in a port of the device for off chip analysis using confocal fluorescence microscopy. Cells in the MFN can also be fixed using a solution of formaldehyde for further analysis after disassembly of the MFN and Si lid. This microfluidic device offers a simple, flexible, and powerful method for depositing two cell populations in separate chambers and may help investigating pathways between the cells populations. PMID- 20013312 TI - Development and validation of a motion and loading system for a rat knee joint in vivo. AB - The influence of biomechanical stimuli on modulating cartilage homeostasis is well recognized. However, many aspects of cellular mechanotransduction in cartilage remain unknown. We developed a computer-controlled joint motion and loading system (JMLS) to study the biological response of cartilage under well characterized mechanical loading environments. The JMLS was capable of controlling (i) angular displacement, (ii) motion frequency, (iii) magnitude of the axial compressive load applied to the moving joint, and it featured real-time monitoring. The accuracy and repeatability of angular position measurements, the kinematic misalignment error as well as the repositioning error of the JMLS were evaluated. The effectiveness of the JMLS in implementing well-defined loading protocols such as moderate Passive Motion Loading (PML) and increased Compressive Motion Loading (CML) were tested. The JMLS demonstrated remarkable accuracy and reliability for the measurement and kinematics tests. Moreover, the effectiveness test demonstrated the ability of the JMLS to produce an effective stimulus via PML that led to the suppression of the catabolic effects of immobilization. Interestingly, the biological response of the CML group was catabolic and exhibited a pattern similar to that observed in the immobilization group. This novel non-invasive system may be useful for joint biomechanics studies that require different treatment conditions of load and motion in vivo. PMID- 20013314 TI - State of the Journal 2010: managing growth, extending our reach--now the official journal for the Society of Surgical Oncology and the American Society of Breast Surgeons. PMID- 20013315 TI - Morphine and rapid disease progression in nonhuman primate model of AIDS: inverse correlation between disease progression and virus evolution. AB - HIV and simian immunodeficiency virus (SIV) have a formidable capacity for mutation and adaptation, a characteristic that has contributed to the extensive genetic variability. Evolutionary pressures imposed within the host and the viral capacity to mutate lead to the generation of such variants. To date, very little information is available regarding the evolution of HIV with drug abuse as a cofounding factor. Using our macaque model of drug dependency and AIDS, we have investigated the dynamics of SIV mutations in the genes tat, vpr, envelope, and nef. The results presented in this review, from our laboratory and others, contribute to the overall understanding of how drugs of abuse might influence immune selective pressure contribution to variation in different SIV genes. Additionally, the studies presented could help enlighten the development of HIV vaccines that take into consideration viral diversity. PMID- 20013316 TI - TRAIL and triptolide: an effective combination that induces apoptosis in pancreatic cancer cells. AB - INTRODUCTION: An emerging therapy in oncology is the induction of apoptotic cell death through anti-death receptor therapy. However, pancreatic cancer is resistant to apoptosis including anti-death receptor therapy. We have previously described how triptolide decreases resistance to apoptosis in pancreatic cancer cells in vitro and in vivo. We hypothesized that triptolide decreases tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) resistance in pancreatic cancer cells. The aim of this study was to evaluate the effects that combined therapy with TRAIL and triptolide have on different parameters of apoptosis. METHODS: Four different pancreatic cancer cell lines were exposed to triptolide, TRAIL, or a combination of both drugs. We assessed the effects that combined therapy with TRAIL and triptolide has on cell viability, apoptosis, caspase-3 and caspase-9 activities, and poly(ADP)-ribose polymerase cleavage. RESULTS: Pancreatic cancer cells were resistant to TRAIL therapy; however, combined therapy with triptolide and TRAIL significantly decreased the cell viability in all the cell lines and increased apoptotic cell death as a result of caspase-3 and caspase-9 activation. CONCLUSIONS: Pancreatic cancer is highly resistant to anti-death receptor therapy, but combined therapy with TRAIL and triptolide is an effective therapy that induces apoptotic cell death in pancreatic cancer cells. PMID- 20013317 TI - Oncocytes, oxyphils, Hurthle, and Askanazy cells: morphological and molecular features of oncocytic thyroid nodules. AB - Our understanding of oncocytic change in thyroid nodules is evaluated in light of the recent progress in understanding the mitochondrial DNA, its mutations, and somatic mutations that affect mitochondrial function. These changes are largely unrelated to the genetic events that result in proliferation and neoplastic transformation of thyroid follicular epithelial cells. The criteria for diagnosing lesions that are composed predominantly of oncocytic cells are the same as those applied to follicular lesions that do not contain oncocytic cells, including follicular variant papillary carcinomas, based on nuclear morphology, immunohistochemical profiles, and molecular markers. PMID- 20013318 TI - Protective effects of propolis and related polyphenolic/flavonoid compounds against toxicity induced by irinotecan. AB - Despite the excellent chemotherapeutic effect of irinotecan, its cytotoxicity and genotoxicity in normal cells remains a major problem in chemotherapy. This study was carried out to find whether propolis preparations and related flavonoids (quercetin, naringin) might enhance irinotecan-induced cytotoxicity to tumor cells in mice bearing Ehrlich ascites tumors (EAT) while protecting normal blood, liver, and kidney cells. The preparation of propolis and their flavonoids were given to mice intraperitoneally at a dose of 100 mg kg(-1) body weight for three consecutive days before the ip injection of EAT cells (2*10(6)). Irinotecan was administered ip at dose of 50 mg kg(-1) on days 3, 4, and 5 after tumor cell inoculation. The combination treatment resulted in substantial inhibition of the growth of EAT cells as well as treatment with quercetin or irinotecan alone, whereas other treatment by itself showed little effect. However, when mice were pre-treated with test components prior to irinotecan, the frequencies of irinotecan-induced micronuclei (MN) was decreased but in mice bearing tumor QU and EEP increased number of micronucleated cells. Propolis preparation and related flavonoids were found to exhibit an important immunomodulatory effect and could decrease irinotecan-induced toxic and genotoxic effects to normal cells without effecting irinotecan cytotoxicity in EAT cells. PMID- 20013319 TI - Soluble CD138 serum levels are not associated with other poor prognostic markers in patients with B-chronic lymphocytic leukaemia. AB - The clinical course of CLL is highly variable, and survival from the time of diagnosis of CLL can range from months to decades. Novel biological markers such as IgVH mutation, CD38, and ZAP-70 expression have shown to offer important prognostic informations. Few reports deal with the sCD138 levels and bad prognostic factors in patients with CLL, and contrasting data are reported in literature. In our study, we evaluated the serum level of sCD138 in patients with B-CLL and its relationship with other prognostic markers. There was a significant association between advanced Rai stage and serum sCD138 levels in CLL subjects. Patients with Rai stage III-IV had significantly higher levels of sCD138 with respect to controls (48.85+/-34 ng/ml vs. 31.1+/-19.34 ng/ml; P<0.05). We were unable to demonstrate a significant association between sCD138 serum levels and IgVH gene status, ZAP-70 expression, CD38 expression, beta-2 microglobulin, absolute peripheral blood lymphocytosis, haemoglobin or LDH levels. Our finding that high sCD138 serum levels correlates with advanced stages in patients with B CLL is consistent with the possibility molecule can identify patients with high tumour burden, but the lack of correlation between sCD138 serum levels and markers such the mutation status of IgVH, ZAP-70, and CD38 suggests that sCD138 levels only reflect the clinical stage of disease than the clinical course or progression. PMID- 20013320 TI - The association of serum adiponectin levels with histopathological variables in gastric cancer patients. AB - Adiponectin is a peptide hormone secreted from the adipose tissue, affecting the proliferation and insulin sensitivity in different cell types. The levels of adiponectin have been found to be decreased in hyperinsulinemia and insulin resistant states, such as obesity. The previous studies have suggested that plasma adiponectin levels are decreased in patients with endometrial and breast cancer. In our study, the relationship among serum adiponectin levels, demographic features and histopathological variables was evaluated in gastric cancer patients. Forty consecutive patients with gastric cancer who underwent gastrectomy with standard lymph node dissection were included and 43 healthy controls were included in this study. The serum levels of glucose, insulin, C peptide, HbA1c and adiponectin were measured in both groups. We analyzed the correlation among these parameters and patients' demographic features, such as age, gender, body mass index (BMI) and histopathological variables such as tumor localization, stage, nodal status, histological grade, vascular and lymphatic invasion. The mean age was 60.05+9.72 in patients, while it was 38.6+12.73 in controls. The mean serum adiponectin levels were 12.62+7.9 and 10.07+6.72 ng/ml, respectively, in groups. There was no different in terms of adiponectin, C peptide, HOMA-R level in both groups. On the other hand, BMI, glucose and insulin levels were significantly different in gastric cancer patients in comparison with the controls. There was no correlation among the levels of adiponectin, BMI, insulin and c-peptide levels in patient group (P>0.05). The adiponectin levels of woman were significantly lower than male patients (P=0.002). No relations were detected among tumor stage, tumor localization, nodal status, lymphatic and vascular invasion, and the levels of serum adiponectin (P>0.05). Interestingly, a positive correlation was found between tumor grade and plasma adiponectin levels (r=0.372; P=0.018). Our results suggest that plasma adiponectin levels were similar in both patients with gastric cancer and the controls. In addition, no correlation was found between adiponectin levels and demographic features and histopathological variables of patients. But, in undifferentiated tumors, plasma adiponectin level was significantly higher than well-differentiated grade tumors. PMID- 20013322 TI - Influenza A/pandemic 2009/H1N1 in the setting of allogeneic hematopoietic cell transplantation: a potentially catastrophic problem in a vulnerable population. AB - We describe Influenza A/pandemic 2009/H1N1 in two allogeneic hematopoietic cell transplantation recipients. The main presentation in both cases consisted of flu like symptoms manifesting as, fever, arthralgias and myalgias. The virus was isolated in one case from a throat swab and in another case following a bronchoalveolar lavage. Both patients received oseltamivir at a dose of 75 mg orally twice day. The dose of oseltamivir was increased to 150 mg twice per day due to the lack of improvement or progression of symptoms. In one case, clinical symptoms resolved without sequelae. In the second case, pulmonary symptomatology continued to deteriorate, despite aggressive polymicrobial treatment, requiring mechanical ventilation and ultimately the patient died from respiratory failure. These cases highlight the potentially serious effect of the ongoing Influenza A/pandemic 2009/H1N1 pandemic in this very vulnerable population and the urgent need to establish emergency preparedness strategies by oncology and bone marrow transplantation staff to face this serious healthcare challenge. PMID- 20013321 TI - DNA detective: a review of molecular approaches to wildlife forensics. AB - Illegal trade of wildlife is growing internationally and is worth more than USD$20 billion per year. DNA technologies are well suited to detect and provide evidence for cases of illicit wildlife trade yet many of the methods have not been verified for forensic applications and the diverse range of methods employed can be confusing for forensic practitioners. In this review, we describe the various genetic techniques used to provide evidence for wildlife cases and thereby exhibit the diversity of forensic questions that can be addressed using currently available genetic technologies. We emphasise that the genetic technologies to provide evidence for wildlife cases are already available, but that the research underpinning their use in forensics is lacking. Finally we advocate and encourage greater collaboration of forensic scientists with conservation geneticists to develop research programs for phylogenetic, phylogeography and population genetics studies to jointly benefit conservation and management of traded species and to provide a scientific basis for the development of forensic methods for the regulation and policing of wildlife trade. PMID- 20013323 TI - p53 is critical for the Aurora B kinase inhibitor-mediated apoptosis in acute myelogenous leukemia cells. AB - We previously showed that AZD1152-HQPA, the inhibitor of Aurora B kinase potently induced growth arrest and apoptosis of various types of human leukemia cells including MV4-11 acute myelogenous leukemia (AML) cells, although the molecular mechanisms by which this class of kinase inhibitors induces apoptosis remain to be fully elucidated. We have recently established the MV4-11 subline, designated as MV4-11 TP53 R248W, which possesses transcriptionally inactive R248W mutation in the TP53 gene. MV4-11 TP53 R248W cells were relatively resistant to AZD1152 HQPA-mediated growth arrest, as measured by MTT and clonogenic assays. AZD1152 HQPA (10-100 nM, 48 h) strikingly induced apoptosis of MV4-11 cells, as assessed by Annexin V binding, loss of mitochondrial outer membrane potential, and activation of caspase cascade, in parallel with up-regulation of p53 and its target molecules Bax and Noxa. Notably, AZD1152-HQPA (10-100 nM, 48 h) induced polyploidy rather than apoptosis in MV4-11 TP53 R248W cells. The polyploid cells were eventually eliminated via apoptosis at later time period (72-120 h) in association with up-regulation of p73. Taken together, p53 plays an important role in AZD1152-HQPA-induced growth arrest and early onset of apoptosis in AML cells. P73 may mediate the late onset of apoptosis to eliminate the polyploid cells caused by the inhibitor of Aurora B kinase. PMID- 20013325 TI - Examination of useful items for the assessment of fall risk in the community dwelling elderly Japanese population. AB - OBJECTIVES: The aim of this study was to select useful items for assessing fall risk in healthy elderly Japanese individuals. METHODS: A total of 965 healthy elderly Japanese subjects aged >=60 years (349 males 70.4 +/- 7.1 years, 616 females 69.9 +/- 7.1 years) participated in this study. Of these, 16.6% had suffered from a previous fall. We assumed five fall risk factors: symptoms of falling, physical function, disease and physical symptoms, environment, and behavior and character. Eighty-six items were selected to represent these factors. To confirm the component items for each risk factor, we performed factor analysis (principle factor solution and varimax rotation). The high-fall risk response rate was also calculated for each item, and significant differences in this rate were examined between groups of those who had and not had experienced a fall. RESULTS: Useful items were selected using the following criteria: (1) items showing a significant difference in high fall risk response rate between faller and non-faller groups were selected as useful items; (2) items showing low factor loading (<0.4) for any factor were deleted as inappropriate items; (3) the top two items showing a greater amount of the difference in high fall risk response rate among the representative items for each factor. A total of 50 items were selected from each fall risk factor (symptoms of falling, 3 items; physical function, 22 items; disease and physical symptom, 13 items; environment, 4 items; behavior and character, 8 items). CONCLUSIONS: Based on our results, the selected items can comprehensively assess the fall risk of a healthy elderly Japanese population. In addition, the assessment items for physical function comprised items of different levels of difficulty, and these are able to gradually and comprehensively assess physical function. PMID- 20013324 TI - Activated STAT1 and STAT5 transcription factors in extramedullary hematopoietic tissue in a polycythemia vera patient carrying the JAK2 V617F mutation. AB - The somatic V617F mutation in the Janus kinase (JAK) 2 gene, which causes a valine to phenylalanine substitution at position 617, has recently been found in the majority of patients with polycythemia vera and in many cases with essential thrombocythemia or idiopathic myelofibrosis. Here, we report on a 76-year-old female patient presenting with JAK2V617F-positive polycythemia vera and a pelvic mass with extramedullary hematopoiesis. Immunohistochemistry demonstrated tyrosine phosphorylation of JAK2 kinase as well as STAT1 and STAT5 transcription factors. However, only a minority of the total STAT1 pool was tyrosine phosphorylated and, in contrast to its unphosphorylated counterpart, phospho STAT1 clearly showed nuclear accumulation. While megakaryotes expressed virtually no phospho-STAT1, phosphorylated STAT5 was mainly restricted to megakaryocytes and rarely detected in non-megakaryocytes. Our data suggest that dysregulated STAT signal pathways are engaged in extramedullary hematopoiesis in polycythemia vera. PMID- 20013327 TI - Optical mapping study of blebbistatin-induced chaotic electrical activities in isolated rat atrium preparations. AB - We have studied the spatiotemporal pattern of blebbistatin-induced anomalous electrical activities in isolated rat atrial preparations using the optical mapping of excitation spread. Atrial preparations including the right or left auricle were dissected from adult rat hearts. Each preparation was then stained with a fast merocyanine-rhodanine voltage-sensitive dye (NK2761). Using a multi element (16 x 16) photodiode array, we assessed the spread of excitation optically by timing the initiation of the action potential-related extrinsic absorption changes. The contraction-related optical signals were suppressed by adding (S)-(-)-blebbistatin (10-100 miocroM) to the bathing solution. Blebbistatin had an effective delay time of about 1.5 h following its application, at which time anomalous electrical activities occurred. These took the form of triggered activities and rhythmical spontaneous excitations. We optically mapped the spatiotemporal patterns of the excitation spread during these anomalous electrical activities. When the triggered activities occurred, the site of ectopic focus, where the triggered action potential first appeared, and the area of excitation spread varied in every event. When the rhythmical spontaneous excitations occurred, the excitation spread from the anomalous pacemaker and, occasionally, their spatial shift was observed. In addition, the combination pattern of the spontaneous excitations and triggered activities was also observed. We suggest that these phenomena are due to the disturbed intracellular calcium dynamics induced by the application of blebbistatin. PMID- 20013329 TI - Special announcement: Guidelines to the practice of anesthesia--revised edition 2010. PMID- 20013328 TI - The relationship between exercise intensity and the sweat lactate excretion rate. AB - The purpose of this study was to determine the effect of increases in exercise intensity on the sweat lactate concentration and lactate excretion rate. Eight healthy male volunteers complete a 90-min exercise bout of treadmill walking in a 35 degrees C and 40% relative humidity environmental chamber. During the exercise trial, the subjects performed three 30-min ordered exercise bouts at 60, 70, and 80% of their age-predicted maximum heart rate (HR(max)), with 10 min of rest outside the chamber between bouts. Sweat rate was measured volumetrically during each of the three exercise bouts on the flexor surface of the proximal half of the right forearm. Sweat lactate concentration ([lactate](sweat)) was measured in each sample and multiplied by the forearm sweat rate to calculate the lactate excretion rate (LER). There was a significant (P < 0.05) decrease in the [lactate](sweat) at the 70 and 80% HR(max) exercise intensities compared to the 60% HR(max) exercise intensity. Conversely, the LER increased significantly at the highest two exercise intensities compared to the 60% HR(max) exercise intensity. Such data suggest that increases in exercise intensity require an increase in lactate production, as measured by the LER. Furthermore, the decreased [lactate](sweat) at the higher exercise intensities is most likely the result of increased sweat production causing a dilution effect on the [lactate](sweat), thus limiting its ability to accurately indicate the metabolic activity of the sweat gland. PMID- 20013330 TI - Updates to the standardized reporting guidelines endorsed by the Journal. PMID- 20013331 TI - [Acute compartment syndrome of the forearm after paravasation of contrast medium. Case report]. AB - The paravasation of radiographic contrast medium has a wide spectrum of clinical presentations, ranging from local erythema and swelling to extensive damage to the extremity affected. We report the case of a patient who developed an acute compartment syndrome of the forearm after paravenous mechanical injection of 100 ml radiographic contrast medium. After emergency fasciotomy for muscle and nerve decompression the full range of motion and sensitivity could be restored. PMID- 20013333 TI - [Medicinal therapy for interventional surgery of the peripheral vascular system]. AB - The aim of medicinal treatment, during and after femoral and crural interventions is to prevent early or late onset arterial thrombosis of the treated vascular segments. Therefore, unfractionated heparin is administered during the intervention by an intra-arterial or intravenous approach. To avoid late onset thrombosis, administration of platelet function inhibitors is recommended. However, valid data are only available for acetylsalicylic acid (ASA). Therefore, ASA is recommended for long term medication. In several cardiological studies on stent implantation in coronary vessels the combination of ASA and clopidogrel for dual platelet inhibition has been proven to be effective. These results have been transferred to antithrombotic therapy of the lower extremities despite the lack of dedicated studies. There is no evidence for the use of vitamin K antagonists after peripheral interventions. PMID- 20013335 TI - [Recanalization of the lower leg: PTA or stent?]. AB - Percutaneous transluminal angioplasty (PTA) and stent placement are currently accepted methods for endovascular treatment of critical limb ischemia, if infragenual vessels are involved. Outcome results in high technical success and satisfactory clinical results for treatment of infrapopliteal lesions with regard to patency rates and amputation-free survival. These treatment modalities are also safe for the patients. The question whether PAT alone or additional stent placement is the better choice, is still unanswered due to limited data. PMID- 20013334 TI - [Percutaneous mechanical atherectomy for treatment of peripheral arterial occlusive disease]. AB - Peripheral arterial occlusive disease (PAOD) is still an extremely important politico-economic disease. Diverse treatment procedures exist but the pillars of therapy are changes in lifestyle, such as nicotine abstinence and walking exercise as well as drug therapy. Further therapy options are considered after conventional procedures have been exhausted. These further options consist of improvement of the blood supply by surgical or minimally invasive procedures. The latter therapy options include balloon dilatation and stenting as the most widely used techniques. More recent techniques also used are cryoplasty, laser angioplasty, drug-coated stents or balloons as well as brachytherapy or atherectomy, whereby this list makes no claims to completeness. The multitude of different treatment methods emphatically underlines the fact that no resounding success can be achieved with one single method. The long-term results of both balloon dilatation and stenting techniques show a need for improvement, which elicited the search for additional methods for the treatment of PAOD. Atherectomy represents such an alternative method for treatment of PAOD. Basically, the term atherectomy means the removal of atheroma tissue. For percutaneous atherectomy, in contrast to surgical procedures, it is not necessary to create surgically access to the vessel but accomplishes the atherectomy by means of dedicated systems via a minimally invasive access. There are two basic forms of mechanical atherectomy: directional and rotational systems. PMID- 20013336 TI - The enrichment of TATA box and the scarcity of depleted proximal nucleosome in the promoters of duplicated yeast genes. AB - Population genetic theory of gene duplication suggests that the preservation of duplicate copies requires functional divergence upon duplication. Genes that can be readily modified to produce new gene expression patterns may thus be duplicated often. In yeast, genes exhibit dichotomous expression patterns based on their promoter architectures. The expression of genes that contain TATA box or occupied proximal nucleosome (OPN) tends to be variable and respond to external signals. On the other hand, genes without TATA box or with depleted proximal nucleosome (DPN) are expressed constitutively. We find that recent duplicates in the yeast genome are heavily biased to be TATA box containing genes and not to be DPN genes. This suggests that variably expressed genes, due to the functional organization in their promoters, have higher duplicability than constitutively expressed genes. PMID- 20013337 TI - The mitochondrial genomes of two scallops, Argopecten irradians and Chlamys farreri (Mollusca: Bivalvia): the most highly rearranged gene order in the family Pectinidae. AB - Molluscs in general and bivalves in particular, exhibit an extraordinary degree of mitochondrial gene order variation when compared with other metazoans. Here, we determined the mitochondrial genomes of two scallops Argopecten irradians and Chlamys farreri. The complete mitochondrial genome of A. irradians is 16,211 nts in length and the nearly complete mitochondrial genome of C. farreri is 20,789 nts in length. Both of the genomes contain 35 genes including 12 protein-coding genes, 2 ribosomal RNAs, and 21 transfer RNAs. In contrast to the typical animal mitochondrial genome, both of them lack one protein-coding gene atp8 and two trnSs, but show an additional copy of trnF in A. irradians and of trnM in C. farreri, respectively. Gene order and genome content were compared among the four sequenced scallops. Gene arrangement of C. farreri closely resembles that of Mizuhopecten yessoensis. However, two genomes of C. farreri and A. irradians show only three small identical gene blocks and two genomes of A. irradians and Placopecten magellanicus share only one gene block. Comparison of the gene arrangement demonstrated that the four scallops share few identical gene blocks although they belong to the same family. This feature is seldom observed in Metazoa, even in other molluscan classes. The dramatic gene rearrangement often occurs in bivalves, especially in marine bivalves. In addition, comparisons of genomic character among bivalves are also presented. PMID- 20013338 TI - Overexpression of protein disulfide isomerases enhances secretion of recombinant human transferrin in Schizosaccharomyces pombe. AB - Although the fission yeast Schizosaccharomyces pombe has been used for high-level heterologous protein production, the productivity of secreted human serum transferrin (hTF) has been low, presumably, because the protein harbors twenty disulfide bonds and two N-glycosylation sites. In the present study, we found that overexpression of endogenous putative protein disulfide isomerase (PDI) improved productivity. Whole genome sequence analysis of S. pombe revealed five putative PDI genes and overexpression of two of them, SPAC17H9.14c and SPBC3D6.13c (SpPdi2p or SpPdi3p, respectively), significantly improved the productivity of secreted hTF. GFP-fused SpPdi2p and SpPdi3p were found to localize to the endoplasmic reticulum. Co-overexpression of SpPdi2p or SpPdi3p with hTF coupled with modifications to the growth medium reported in our previous study were able to increase the level of secreted hTF approximately 30-fold relative to conventional conditions. PMID- 20013339 TI - FLO gene-dependent phenotypes in industrial wine yeast strains. AB - Most commercial yeast strains are nonflocculent. However, controlled flocculation phenotypes could provide significant benefits to many fermentation-based industries. In nonflocculent laboratory strains, it has been demonstrated that it is possible to adjust flocculation and adhesion phenotypes to desired specifications by altering expression of the otherwise silent but dominant flocculation (FLO) genes. However, FLO genes are characterized by high allele heterogeneity and are subjected to epigenetic regulation. Extrapolation of data obtained in laboratory strains to industrial strains may therefore not always be applicable. Here, we assess the adhesion phenotypes that are associated with the expression of a chromosomal copy of the FLO1, FLO5, or FLO11 open reading frame in two nonflocculent commercial wine yeast strains, BM45 and VIN13. The chromosomal promoters of these genes were replaced with stationary phase inducible promoters of the HSP30 and ADH2 genes. Under standard laboratory and wine making conditions, the strategy resulted in expected and stable expression patterns of these genes in both strains. However, the specific impact of the expression of individual FLO genes showed significant differences between the two wine strains and with corresponding phenotypes in laboratory strains. The data suggest that optimization of the flocculation pattern of individual commercial strains will have to be based on a strain-by-strain approach. PMID- 20013341 TI - A lifetime of fear of being laughed at: an aged perspective. AB - This paper reviews recent literature on gelotophobia (i.e., the fear of being laughed at) with an emphasis on age-specific aspects. Research with two instruments, the GELOPH and PhoPhiKat questionnaires, is presented with special attention being given to sociodemographic correlates and differences in intelligence, character strengths, personality, emotion, and humor. Quite consistently gelotophobes tend to misread positively motivated smiling and laughter (e.g. in social interactions, photographs or auditorily presented) and have lower values in many, but not all, components of humor. They have a low propensity to joy and a disposition to experience shame and fear. More generally they tend to describe themselves as being introverted and neurotic, and they underestimate their own potential while not actually being less capable. Furthermore, new data are presented suggesting that age-related vulnerabilities may be additional sources of ridicule making gelotophobia more of a problem for the elderly. Finally, the prevalence of this fear over the lifespan and potential cohort effects are discussed. It is concluded that more research into this fear and its adverse impact on social interactions, even humorous ones, of the elderly is needed. PMID- 20013340 TI - Conservation of miR-15a/16-1 and miR-15b/16-2 clusters. AB - MiR-15a/16-1 and miR-15b/16-2 clusters have been shown to play very important roles in regulating cell proliferation and apoptosis by targeting cell cycle proteins and the antiapoptotic Bcl-2 gene. However, the physiological implications of those two clusters are largely elusive. By aligning the primary miR-15a/16-1 sequence among 44 vertebrates, we found that there was a gap in the homologous region of the rat genome. To verify that there was a similar miR 15a/16-1 cluster in rats, we amplified this region from rat genomic DNA using PCR and found that a 697-bp sequence was missing in the current rat genome database, which covers the miR-15a/16-1 cluster. Subsequently, we also investigated the expression pattern of individual miRNAs spliced from miR-15a/16-1 and miR-15b/16 2 clusters, including miR-15a, miR-15a*, miR-15b, miR-15b*, miR-16-1/2, and miR 16-1/2* from various rat tissues, and found that all of those miRNAs were expressed in the investigated tissues. MiR-16 was most expressed in the heart, followed by the brain, lung, kidney, and small intestine, which indicates tissue specificity for individual miRNA expression from both clusters. Our results demonstrated that both miR-15a/16-1 and miR-15b/16-2 clusters are highly conserved among mammalian species. The investigation of the biological functions of those two clusters using transgenic or knockout/knockdown models will provide new clues to understanding their implications in human diseases and finding a new approach for miRNA-based therapy. PMID- 20013344 TI - Intraductal fibroadenoma and intraductal phyllodes tumour--a part of the spectrum of the breast ductal adenoma? PMID- 20013343 TI - Dynamic movements of Ro52 cytoplasmic bodies along microtubules. AB - The RING-finger protein Ro52/TRIM21 is known as an autoantigen and is recognized by anti-Ro/SSA antibodies, which are commonly found in patients with Sjogren's syndrome and systemic lupus erythematosus. Recently, Ro52 has been shown to localize to distinct structures called cytoplasmic bodies and function as an E3 ubiquitin ligase. However, the Ro52 cytoplasmic bodies have not been well characterized. In this study, we investigated the Ro52 cytoplasmic bodies using fluorescence microscopy. This analysis revealed that the Ro52 cytoplasmic bodies are diffusely located in the cytoplasm and exist independently of TRIM5alpha cytoplasmic bodies. Our results further showed that the Ro52 cytoplasmic bodies are not stained with MitoTracker dye and are not colocalized with the proteasome subunit Rpt5, the caveolae component caveolin-1, the endosome markers (EEA1, Rab5, and Rab7), and the lysosome marker LAMP2. These results indicate that the Ro52 cytoplasmic bodies are not mitochondria, proteasome-enriched structures, caveolae, endosomes, or lysosomes. Importantly, the Ro52 cytoplasmic bodies are highly motile and are located along the microtubule network. These results suggest that the Ro52 cytoplasmic bodies are unidentified structures that are transported along the microtubule network. PMID- 20013345 TI - Primary choriocarcinoma of the liver: a clinicopathological study of five cases in males. AB - Primary choriocarcinomas of the liver are rare. Previous reported cases were mostly in infants with only rare adult cases. Here, we presented five adult cases. The patients were all males, with an average age of 41.6 years (from 36 to 48 years). Clinical presentations included right upper abdominal pain or abdominal distension. All patients presented with a large hepatic mass on ultrasonography that measured 11 cm on average in the greatest diameter. Elevated serum human chorionic gonadotropin (HCG) levels were noted in all cases. At presentation, the tumor was confined to the liver in two patients and therefore surgically resected. The other three patients presented with extrahepatic metastases on imaging study and therefore only received chemotherapy. All five patients died from the tumor within 2 to 8 months. Autopsy was performed for all five cases. The autopsy confirmed that the choriocarcinoma was confined to the liver in two surgically resected cases. The other three patients had metastatic choriocarcinoma in the lung (two patients), peritoneum (one patient), adrenal glands (one patient), and brain (one patient). None of the patients had any evidence of a testicular tumor or scar after examination of the entirely submitted testes. No tumor was observed in central nervous system, mediastinum, or other organs other than described above. Grossly, the primary tumors were large, soft, hemorrhagic, and with foci of necrosis. Histologically, the tumors were composed of mononucleated trophoblastic cells with round nuclei, clear cytoplasm, and prominent nucleoli admixed with large, multinucleated syncytiotrophoblastic cells. Immunohistochemically, tumor cells were strongly positive for keratin, HCG, and focally positive for human placental lactogen. Ki 67 proliferation index was high (mean 75%) in the mononucleated trophoblastic cells. Our series is the largest one to document primary hepatic choriocarcinoma in adults. Although these tumors are rare, they behave in a very aggressive fashion. PMID- 20013346 TI - Clinicopathologic characteristics of pleomorphic carcinoma of the breast. AB - Pleomorphic carcinoma of the breast is considered a rare variant of high-grade ductal NOS carcinoma (NOS-IDC), and the prognosis is poor. However, its clinicopathologic features are not well-characterized. Using the criteria delineated in the World Health Organization breast tumor classification of 2003, ten cases of pleomorphic carcinoma were identified from 9794 NOS-IDC in our archived materials that were originally diagnosed as high-grade infiltrating ductal carcinoma of breast. To investigate the clinicopathologic characteristics and to elucidate the histologic diagnosis and differential diagnosis of this entity, we reviewed the pathology manifestations and clinical features of these cases and examined the tumor expression of ER, PR, PCNA, AE(1)/AE(3), p53, S-100, C-erbB-2, EMA, p63, and Bcl-2 by immunohistochemistry. PMID- 20013348 TI - Cell death induced by novel fluorinated taxanes in drug-sensitive and drug resistant cancer cells. AB - The aim of this study is to compare the effects of new fluorinated taxanes SB-T 12851, SB-T-12852, SB-T-12853, and SB-T-12854 with those of the classical taxane paclitaxel and novel non-fluorinated taxane SB-T-1216 on cancer cells. Paclitaxel sensitive MDA-MB-435 and paclitaxel-resistant NCI/ADR-RES human cancer cell lines were used. Cell growth and survival evaluation, colorimetric assessment of caspases activities, flow cytometric analyses of the cell cycle and the assessment of mitochondrial membrane potential, reactive oxygen species (ROS) and the release of cytochrome c from mitochondria were employed. Fluorinated taxanes have similar effects on cell growth and survival. For MDA-MB-435 cells, the C(50) of SB-T-12851, SB-T-12852, SB-T-12853 and SB-T-12854 was 3 nM, 4 nM, 3 nM and 5 nM, respectively. For NCI/ADR-RES cells, the C(50) of SB-T-12851, SB-T-12852, SB T-12853, and SB-T-12854 was 20 nM, 20 nM, 10 nM and 10 nM, respectively. Selected fluorinated taxanes, SB-T-12853 and SB-T-12854, at the death-inducing concentrations (30 nM for MDA-MB-435 and 300 nM for NCI/ADR-RES) were shown to activate significantly caspase-3, caspase-9, caspase-2 and also slightly caspase 8. Cell death was associated with significant accumulation of cells in the G(2)/M phase. Cytochrome c was not released from mitochondria and other mitochondrial functions were not significantly impaired. The new fluorinated taxanes appear to use the same or similar mechanisms of cell death induction as compared with SB-T 1216 and paclitaxel. New fluorinated and non-fluorinated taxanes are more effective against drug-resistant cancer cells than paclitaxel. Therefore, new generation of taxanes, either non-fluorinated or fluorinated, are excellent candidates for further and detailed studies. PMID- 20013350 TI - An examination of the reciprocal relationships between adolescents' aggressive behaviors and their perceptions of parental nurturance. AB - This study examined reciprocal relationships between adolescents' perceptions of parental nurturance and two types of adolescent aggressive behaviors (indirect and direct aggression) using a transactional model. Three waves of longitudinal data were drawn from the Canadian National Longitudinal Survey of Children and Youth. The sample included 1,416 (735 female) adolescents who were 10- and 11 year-olds at Time 1 and became 14-and 15-year-olds at Time 3. The findings failed to support reciprocal effects, but confirmed parental effects at different ages for girls and boys. For girls, perceptions of parental nurturance at age 10 were negatively associated with both indirect and direct aggression at age 12. For boys, perceptions of parental nurturance at age 12 were negatively associated with both aggressive behaviors at age 14. Future research should continue to investigate reciprocal effects in parent-adolescent relationships to identify developmental periods where the effect of adolescents' or their parents' behavior may be stronger. PMID- 20013349 TI - Hypoxia-targeting by tirapazamine (TPZ) induces preferential growth inhibition of nasopharyngeal carcinoma cells with Chk1/2 activation. AB - Hypoxia is commonly developed in solid tumors, which contributes to metastasis as well as radio- and chemo-resistance. Nasopharyngeal carcinoma (NPC) is a highly invasive and metastatic head and neck cancer prevalent in Southeast Asia with a high incidence rate of 15-30/100,000 persons/year (comparable to that of pancreatic cancer in the US). Previous clinical studies in NPC showed that hypoxia is detected in almost 100% of primary tumors and overexpression of hypoxia markers correlated with poor clinical outcome. Tirapazamine (TPZ) is a synthetic hypoxia-activated prodrug, which preferentially forms cytotoxic and DNA damaging free radicals under hypoxia, thus selectively eradicate hypoxic cells. Here, we hypothesized that specific hypoxia-targeting by this clinical trial agent may be therapeutic for NPC. Our findings demonstrated that under hypoxia, TPZ was able to induce preferential growth inhibition of NPC cells, which was associated with marked cell cycle arrest at S-phase and PARP cleavage (a hallmark of apoptosis). Examination of S-phase checkpoint regulators revealed that Chk1 and Chk2 were selectively activated by TPZ in NPC cells under hypoxia. Hypoxia selectivity of TPZ was also demonstrated by preferential downregulation of several important hypoxia-induced markers (HIF-1alpha, CA IX and VEGF) under hypoxia. Furthermore, we demonstrated that TPZ was equally effective and hypoxia selective even in the presence of the EBV oncoprotein, LMP1 or the EBV genome. In summary, encouraging results from this proof-of-concept study implicate the therapeutic potential of hypoxia-targeting approaches for the treatment of NPC. PMID- 20013351 TI - A multi-level examination of the association between older social models in the school environment and overweight and obesity among younger students. AB - Few studies have examined how school characteristics are associated with obesity. The purpose of this study was to examine the student- and school-level characteristics associated with being overweight and obese among 12,049 students in grades 9 and 10 (junior students) attending 76 secondary schools in Ontario, Canada. The sample was 49.2% female and the average age was 14.7 years. Overall, 15.0% of the sample was considered overweight and 6.3% of the sample was considered obese. Multi-level regression analysis revealed significant between school random variation for the likelihood of a junior student (grade 9 or 10) being overweight or obese. For each 1% increase in the prevalence of obese senior students (grade 11 and 12) at a school, the odds of a junior student at that school being overweight or obese increased significantly. Important student-level characteristics included physical activity, screen time sedentary behaviour (e.g., watching television), participation in varsity sports and gender. Future research should evaluate if the optimal population level impact for school-based obesity prevention programming might be achieved by targeting the schools that are putting students at the greatest risk. PMID- 20013352 TI - Glycosaminoglycans from earthworms (Eisenia andrei). AB - The whole tissue of the earthworm (Eisenia andrei) was lyophilized and extracted to purify glycosaminoglycans. Fractions, eluting from an anion-exchange column at 1.0 M and 2.0 M NaCl, showed the presence of acidic polysaccharides on agarose gel electrophoresis. Monosaccharide compositional analysis showed that galactose and glucose were most abundant monosaccharides in both fractions. Depolymerization of the polysaccharide mixture with glycosaminoglycan-degrading enzymes confirmed the presence of chondroitin sulfate/dermatan sulfate and heparan sulfate in the 2.0 M NaCl fraction. The content of GAGs (uronic acid containing polysaccharide) in the 2.0 M NaCl fraction determined by carbazole assay was 2%. Disaccharide compositional analysis using liquid chromatography electrospray ionization mass spectrometry (LC-ESI-MS) analysis after chondroitinase digestion (ABC and ACII), showed that the chondroitin sulfate/dermatan sulfate contained a 4-O-sulfo (76%), 2,4-di-O-sulfo (15%), 6-O sulfo (6%), and unsulfated (4%) uronic acid linked N-acetylgalactosamine residues. LC-ESI-MS analysis of heparin lyase I/II/III digests demonstrated the presence of N-sulfo (69%), N-sulfo-6-O-sulfo (25%) and 2-O-sulfo-N-sulfo-6-O sulfo (5%) uronic acid linked N-acetylglucosamine residues. PMID- 20013353 TI - Effects of flooding on leaf development, transpiration, and photosynthesis in narrowleaf cottonwood, a willow-like poplar. AB - The narrowleaf cottonwood, Populus angustifolia, occurs in occasionally flooded, low elevation zones along river valleys near the North American Rocky Mountains. This small poplar has narrow leaves and fine branching and thus resembles willows, which are commonly flood-tolerant. We investigated the flood response of narrowleaf cottonwoods and a related native hybrid, jackii cottonwood (P. x jackii = P. balsamifera x P. deltoides), by studying saplings of 24 clones in a greenhouse, with some pots being inundated to provide the flood treatment. Flooding slightly reduced leaf numbers (-10%), and leaf sizes were reduced by about 21% in female P. angustifolia versus a 50% reduction in the female hybrids. Flooding-reduced stomatal conductance and net photosynthetic rate, and reduced transpiration particularly in P. x jackii. The effects on foliar gas exchange declined over a 5-week interval, suggesting compensation. The moderate impact of flooding supports the hypothesis that narrowleaf cottonwoods are flood-tolerant, and we anticipate that these trees could provide traits to increase the flood tolerance of fast-growing hybrid poplars. The results further indicate that female cottonwoods may be more flood-tolerant than males, and females could be more successful in lower, flood-prone sites. PMID- 20013354 TI - The long-time dynamics of two hydrodynamically-coupled swimming cells. AB - Swimming microorganisms such as bacteria or spermatozoa are typically found in dense suspensions, and exhibit collective modes of locomotion qualitatively different from that displayed by isolated cells. In the dilute limit where fluid mediated interactions can be treated rigorously, the long-time hydrodynamics of a collection of cells result from interactions with many other cells, and as such typically eludes an analytical approach. Here, we consider the only case where such problem can be treated rigorously analytically, namely when the cells have spatially confined trajectories, such as the spermatozoa of some marine invertebrates. We consider two spherical cells swimming, when isolated, with arbitrary circular trajectories, and derive the long-time kinematics of their relative locomotion. We show that in the dilute limit where the cells are much further away than their size, and the size of their circular motion, a separation of time scale occurs between a fast (intrinsic) swimming time, and a slow time where hydrodynamic interactions lead to change in the relative position and orientation of the swimmers. We perform a multiple-scale analysis and derive the effective dynamical system--of dimension two--describing the long-time behavior of the pair of cells. We show that the system displays one type of equilibrium, and two types of rotational equilibrium, all of which are found to be unstable. A detailed mathematical analysis of the dynamical systems further allows us to show that only two cell-cell behaviors are possible in the limit of t-->infinity, either the cells are attracted to each other (possibly monotonically), or they are repelled (possibly monotonically as well), which we confirm with numerical computations. Our analysis shows therefore that, even in the dilute limit, hydrodynamic interactions lead to new modes of cell-cell locomotion. PMID- 20013355 TI - Emergent group dynamics governed by regulatory cells produce a robust primary T cell response. AB - The currently accepted paradigm for the primary T cell response is that effector T cells commit to autonomous developmental programs. This concept is based on several experiments that have demonstrated that the dynamics of a T cell response is largely determined shortly after antigen exposure and that T cell dynamics do not depend on the level and duration of antigen stimulation. Another experimental study has also shown that T cell responses are robust to variations in antigen specific precursor frequency. Various mathematical models have corroborated the first result that programmed T cell responses are insensitive to the level of antigen stimulation. However, this paper proposes that programmed responses do not entirely explain the robustness of T cell dynamics to variations in precursor frequency. This work studies the hypothesis that the dynamics of a T cell response may also be governed by a feedback loop involving adaptive regulatory cells rather than by intrinsic developmental programs. We formulate two mathematical models based on T cell developmental programs. In one model, effector cells undergo a fixed number of divisions before dying. In the second model, effector cells live for a fixed time during which they may divide. The study of these models suggests that developmental programs are not sufficiently robust as they produce an immune response that directly scales with precursor frequencies. Consequently, we derive a third model based on the principle that adaptive regulatory T cells develop in the course of an immune response and suppress effector cells. Our simulations show that this feedback mechanism responds robustly over a range of at least four orders of magnitude of precursor frequencies. We conclude that the proliferation program paradigm does not entirely capture the observed robustness of T cell responses to variations in precursor frequency. We propose an alternative mechanism by which the primary T cell response is governed by an emergent group dynamic and not by individual T cell programs. PMID- 20013356 TI - Trace metal release from orthodontic appliances by in vivo studies: a systematic literature review. AB - The paper discusses various approaches used to investigate biocompatibility by the analysis of metals release by the materials of which orthodontic appliances are made. Analysis of various biomarkers of exposure: saliva, serum, mucosa cells, or urine is used in in vivo tests. In this work, the techniques, results, and conclusions of original papers were compared by the implementation of the concept of a systematic review. The aim of the present work was to report the state-of-the-art in the research on methods used to assess exposure to trace metals from orthodontic appliances. The PubMed search identified 35 studies, among which nine met the selection criteria. The general conclusion in the studies was that metal ions were released mostly in the initial stage of the treatment. However, the majority of studies included 1-2 months long period and did not reflect long-term changes nor the impact of the complete treatment, the duration of which is several years, on the whole organism and the overall accumulation of metals from orthodontic appliances. In studies which evaluated nickel concentrations in blood and urine, long-term metal release was detected and significant differences were found. It leads to the conclusion that nickel ions are released from orthodontic appliances in measurable amounts to human organism. PMID- 20013357 TI - The effects of feeding clinoptilolite on hematology, performance, and health of newborn lambs. AB - The effects of feeding clinoptilolite on hematology, performance, and health of newborn Balouchi lambs were evaluated in this experiment. In a completely randomized design, 30 newborn lambs were allocated to three groups and fed by basal diet (C0; without clinoptilolite) and C1 and C2 (the basal diet plus 1.5% and 3% clinoptilolite, respectively, for 6 weeks (3 weeks before and 3 weeks after weaning)). Blood samples were taken from all lambs, at the time when the animals were allocated to the experimental diet and at the end of each week of experiment, and analyzed for hematology, plasma fibrinogen, and total protein. Performance and health of all lambs were measured. Fecal consistency score and diarrhea severity were evaluated. There was no difference between lambs in case of hematological parameters. Lambs fecal consistency score and severity of diarrhea were lowest (P < 0.05) for lambs on C1 and C2 and highest for lambs on C0. Dry matter intake and feed conservation ratio were similar between the groups of lambs fed by different diets, but daily gain of lambs differed significantly (P < 0.05) and was higher in C2. It was concluded that addition of 3% clinoptilolite to starter diet of newborn lamb can reduce incidence and severity of diarrhea, although its effect on hematology and performance was negligible. PMID- 20013358 TI - Correlation analysis of measurement result between accelerator mass spectrometry and gamma counter. AB - OBJECTIVE: The guidelines for microdosing in clinical trials were published in Japan in 2008 following the guidelines of the European Medicines Agency and the Food and Drug Administration. They recommend utilizing accelerator mass spectrometry (AMS) and positron emission tomography as candidates for monitoring drug metabolites in preclinical studies. We correlate the two methods by measuring appropriately labeled tissue samples from various mouse organs using both AMS and gamma counter. METHODS: First, we measured the (14)C background levels in mouse organs using the AMS system. We then clarified the relationship between AMS and gamma counter by simultaneously administering (14)C-2-fluoro-2 deoxyglucose ((14)C-FDG) and (18)F-2-fluoro-2-deoxyglucose ((18)F-FDG). Tissue distribution was examined after 30 min, 1 h, 2 h and 4 h using the AMS system for (14)C-FDG and gamma counter for (18)F-FDG. Background (14)C levels were subtracted from the data obtained with radiotracer administration. RESULTS: The background (14)C concentration differed with tissue type measured. Background (14)C concentration in mouse liver was higher than in other organs, and was approximately 1.5-fold that in blood. The correlation coefficient (r) of the measurements between AMS ((14)C-FDG) and gamma counter ((18)F-FDG) was high in both normal (0.99 in blood, 0.91 in brain, 0.61 in liver and 0.78 in kidney) and tumor-bearing mice (0.95 in blood and 0.99 in tumor). The clearance profile of (18)F-FDG was nearly identical to that of (14)C-FDG measured with AMS. CONCLUSIONS: Accelerator mass spectrometry analysis has an excellent correlation with biodistribution measurements using gamma counter. Our results suggest that the combination of AMS and PET can act as a complementary approach to accelerate drug development. PMID- 20013359 TI - Incidental finding of an 11C-acetate PET-positive multiple myeloma. AB - Multiple myeloma is a malignancy of plasma cells. The (18)F-FDG PET findings of multiple myeloma have been reported previously. However, the (11)C-acetate PET findings have not been clarified. Here, we report a case of multiple myeloma detected with (11)C-acetate PET in a 51-year-old male patient with known hepatocellular carcinoma. The patient was admitted for management of a pathologic fracture of the right tibia. Imaging workup including X-ray, magnetic resonance image, bone scintigraphy; (18)F-FDG led to a suspicion of metastatic bony lesions. Further, these lesions showed increased uptake on (11)C-acetate PET. Wide excision of the right tibia was performed, and histopathological examination of the lesion confirmed multiple myeloma. This case illustrates the characteristic (11)C-acetate PET findings of multiple myeloma. PMID- 20013360 TI - An introduction to proteome bioinformatics. AB - This book is part of the Methods in Molecular Biology series, and provides a general overview of computational approaches used in proteome research. In this chapter, we give an overview of the scope of the book in terms of current proteomics experimental techniques and the reasons why computational approaches are needed. We then give a summary of each chapter, which together provide a picture of the state of the art in proteome bioinformatics research. PMID- 20013361 TI - Bioinformatics methods for protein identification using Peptide mass fingerprinting. AB - Protein identification by mass spectrometry (MS) is an important technique in proteomics. By searching an MS spectrum against a given protein database, the most matched proteins are sorted using a scoring function and the top one is often considered the correctly identified protein. Peptide mass fingerprinting (PMF) is one of the major methods for protein identification using MS technology. It is faster and cheaper than the other popular technique - Tandem Mass Spectrometry. Key bioinformatics issues in PMF analysis include designing a scoring function to quantitatively measure the degree of consistency between a PMF spectrum and a protein sequence and assessing the confidence of identified proteins. In this chapter, we will introduce several scoring functions that were developed by others and us. We will also provide a new statistic model to evaluate the confidence of the score and make an improvement for ranking proteins in protein identification. Our developments have been implemented in a software package "ProteinDecision," which is available at http://digbio.missouri.edu/ProteinDecision/ . PMID- 20013362 TI - Computational approaches to peptide identification via tandem MS. AB - The peptide identification problem lies at the heart of modern proteomic methodology, from which the presence of a particular protein or proteins in a sample may be inferred. The challenge is to find the most likely amino acid sequence, which corresponds to each tandem mass spectrum that has been collected, and produce some kind of score and associated statistical measure that the putative identification is correct. This approach assumes that the peptide (and parent protein) sequence in question is known and is present in the database which is to be searched, as opposed to de novo methods, which seek to identify the peptide ab initio. This chapter will provide an overview of the methods that common, popular software tools employ to search protein sequence databases to provide the non-expert reader with sufficient background to appreciate the choices they can make. This will cover the approaches used to compare experimental and theoretical spectra and some of the methods used to validate and provide higher confidence in the assignments. PMID- 20013363 TI - Scoring and validation of tandem MS peptide identification methods. AB - A variety of methods are described in the literature to assign peptide sequences to observed tandem MS data. Typically, the identified peptides are associated only with an arbitrary score that reflects the quality of the peptide-spectrum match but not with a statistically meaningful significance measure. In this chapter, we discuss why statistical significance measures can simplify and unify the interpretation of MS-based proteomic experiments. In addition, we also present available software solutions that convert scores into sound statistical measures. PMID- 20013364 TI - Target-decoy search strategy for mass spectrometry-based proteomics. AB - Accurate and precise methods for estimating incorrect peptide and protein identifications are crucial for effective large-scale proteome analyses by tandem mass spectrometry. The target-decoy search strategy has emerged as a simple, effective tool for generating such estimations. This strategy is based on the premise that obvious, necessarily incorrect "decoy" sequences added to the search space will correspond with incorrect search results that might otherwise be deemed to be correct. With this knowledge, it is possible not only to estimate how many incorrect results are in a final data set but also to use decoy hits to guide the design of filtering criteria that sensitively partition a data set into correct and incorrect identifications. PMID- 20013365 TI - Understanding and exploiting Peptide fragment ion intensities using experimental and informatic approaches. AB - Tandem mass spectrometry is a widely used tool in proteomics. This section will address the properties that describe how protonated peptides fragment when activated by collisions in a mass spectrometer and how that information can be used to identify proteins. A review of the mobile proton model is presented, along with a summary of commonly observed peptide cleavage enhancements, including the proline effect. The methods used to elucidate peptide dissociation chemistry by using both small groups of model peptides and large datasets are also discussed. Finally, the role of peak intensity in commercially available and developmental peptide identification algorithms is examined. PMID- 20013366 TI - Spectral library searching for peptide identification via tandem MS. AB - Spectral library searching is a new approach in proteomic data analysis that promises to address some of the shortcomings of sequence database searching, currently the dominant method for inferring peptide identifications from tandem mass spectra. In spectral searching, a spectral library is first meticulously compiled from a large collection of previously observed and identified peptide MS/MS spectra. The unknown spectrum can then be identified by comparing it to all the candidates in the spectral library for the best match. It offers the benefits of tremendous speed gain and increase in sensitivity and selectivity, compared to sequence searching. This article provides a concise roadmap for the proteomics researchers to start using spectral library searching in their data analysis workflow. PMID- 20013367 TI - De novo sequencing methods in proteomics. AB - The review describes methods of de novo sequencing of peptides by mass spectrometry. De novo methods utilize computational approaches to deduce the sequence or partial sequence of peptides directly from the experimental MS/MS spectra. The concepts behind a number of de novo sequencing methods are discussed. The other approach to identify peptides by tandem mass spectrometry is to match the fragment ions with virtual peptide ions generated from a genomic or protein database. De novo methods are essential to identify proteins when the genomes are not known but they are also extremely useful even when the genomes are known since they are not affected by errors in a search database. Another advantage of de novo methods is that the partial sequence can be used to search for posttranslation modifications or for the identification of mutations by homology based software. PMID- 20013368 TI - Cross species proteomics. AB - Proteomics has advanced in leaps and bounds over the past couple of decades. However, the continuing dependency of mass spectrometry-based protein identification on the searching of spectra against protein sequence databases limits many proteomics experiments. If there is no sequenced genome for a given species, then cross species proteomics is required, attempting to identify proteins across the species boundary, typically using the sequenced genome of a closely related species. Unlike sequence searching for homologues, the proteomics equivalent is confounded by small differences in amino acid sequences, leading to large differences in peptide masses; this renders mass matching of peptides and their product ions difficult. Therefore, the phylogenetic distance between the two species and the attendant level of conservation between the homologous proteins play a huge part in determining the extent of protein identification that is possible across the species boundary. In this chapter, we review the cross species challenge itself, as well as various approaches taken to deal with it and the success met with in past studies. This is followed by recommendations of best practice and suggestions to researchers facing this challenge as well as a final section predicting developments, which may help improve cross species proteomics in the future. PMID- 20013369 TI - Gene model detection using mass spectrometry. AB - The utility of a genome sequence in biological research depends entirely on the comprehensive description of all of its functional elements. Analysis of genome sequences is still predominantly gene-centric (i.e., identifying gene models/open reading frames). In this article, we describe a proteomics-based method for identifying open reading frames that are missed by computational algorithms. Mass spectrometry-based identification of peptides and proteins from biological samples provide evidence for the expression of the genome sequence at the protein level. This proteogenomic annotation method combines computationally predicted ORFs and the genome sequence with proteomics to identify novel gene models. We also describe our proteogenomic mapping pipeline - a set of computational tools that automate the proteogenomic annotation work flow. This pipeline is available for download at www.agbase.msstate.edu/tools/ . PMID- 20013370 TI - Signal processing in proteomics. AB - Computational proteomics applications are often imagined as a pipeline, where information is processed in each stage before it flows to the next one. Independent of the type of application, the first stage invariably consists of obtaining the raw mass spectrometric data from the spectrometer and preparing it for use in the later stages by enhancing the signal of interest while suppressing spurious components. Numerous approaches for preprocessing MS data have been described in the literature. In this chapter, we will describe both, standard techniques originating from classical signal and image processing, and novel computational approaches specifically tailored to the analysis of MS data sets. We will focus on low level signal processing tasks such as baseline reduction, denoising, and feature detection. PMID- 20013371 TI - A high-performance reconfigurable computing solution for Peptide mass fingerprinting. AB - High-throughput, MS-based proteomics studies are generating very large volumes of biologically relevant data. Given the central role of proteomics in emerging fields such as system/synthetic biology and biomarker discovery, the amount of proteomic data is expected to grow at unprecedented rates over the next decades. At the moment, there is pressing need for high-performance computational solutions to accelerate the analysis and interpretation of this data.Performance gains achieved by grid computing in this area are not spectacular, especially given the significant power consumption, maintenance costs and floor space required by large server farms.This paper introduces an alternative, cost effective high-performance bioinformatics solution for peptide mass fingerprinting based on Field Programmable Gate Array (FPGA) devices. At the heart of this approach stands the concept of mapping algorithms on custom digital hardware that can be programmed to run on FPGA. Specifically in this case, the entire computational flow associated with peptide mass fingerprinting, namely raw mass spectra processing and database searching, has been mapped on custom hardware processors that are programmed to run on a multi-FPGA system coupled with a conventional PC server. The system achieves an almost 2,000-fold speed-up when compared with a conventional implementation of the algorithms in software running on a 3.06 GHz Xeon PC server. PMID- 20013372 TI - Mining proteomic MS/MS data for MRM transitions. AB - Multiple reaction monitoring (MRM) of peptides is a popular proteomics technique that employs tandem mass spectrometry to quantify selected proteins of interest, such as those previously identified in differential protein identification studies. Using this technique, the specificity of precursor to product transitions is exploited to determine the absolute quantity of multiple proteins in a single sample. Selection of suitable transitions is critical for the success of MRM experiments, but accurate theoretical prediction of fragmentation patterns and peptide signal intensity is currently not possible. A recently proposed solution to this problem is to combine knowledge of the preferred properties of transitions for MRM, taken from expert practitioners, with MS/MS evidence extracted from a proteomics data repository. In addition, by predicting retention time for each peptide candidate, it allows selection of several compatible transition candidates that can be monitored simultaneously, permitting MRM. In this chapter, we explain how to go about designing transitions using the web based transition design tool, MRMaid, which leverages high quality MS/MS evidence from the Genome Annotating Proteomic Pipeline (GAPP). PMID- 20013373 TI - OpenMS and TOPP: open source software for LC-MS data analysis. AB - The automatic analysis of mass spectrometry data is becoming more and more important since increasingly larger datasets are readily available that cannot be evaluated manually. This has triggered the development of several open-source software libraries for the automatic analysis of such data. Among those is OpenMS together with TOPP (The OpenMS Proteomics Pipeline). OpenMS is a C++ library for rapid prototyping of complex algorithms for the analysis of mass spectrometry data. Based on the OpenMS library, TOPP provides a collection of tools for the most important tasks in proteomics analysis. The tight coupling of OpenMS and TOPP makes it easy to extend TOPP by adding new tools to the OpenMS library. We describe the overall concepts behind the software and illustrate its use with several examples. PMID- 20013374 TI - Trans-proteomic pipeline: a pipeline for proteomic analysis. AB - Mass spectrometry has quickly become an essential tool in molecular biology laboratories. Here, we describe the Trans-Proteomic Pipeline, a collection of software tools, to facilitate the analysis, exchange, and comparison of MS data. The pipeline is instrument-independent and supports most commonly used proteomics workflows, including quantitative applications such as ICAT, iTRAQ, and SILAC. Importantly, the pipeline uses open, standard data formats and calculates accurate estimates of sensitivity and error rates, thus allowing for meaningful data exchange. In this chapter, we will introduce the various components of the pipeline in the context of three typical proteomic use-case scenarios. PMID- 20013376 TI - Automated generic analysis tools for protein quantitation using stable isotope labeling. AB - Isotope labeling combined with LC-MS/MS provides a robust platform for quantitative proteomics. Protein quantitation based on mass spectral data falls into two categories: one determined by MS/MS scans, e.g., iTRAQ-labeling quantitation, and the other by MS scans, e.g., quantitation using SILAC, ICAT, or (18)O labeling. In large-scale LC-MS proteomic experiments, tens of thousands of MS and MS/MS spectra are generated and need to be analyzed. Data noise further complicates the data analysis. In this chapter, we present two automated tools, called Multi-Q and MaXIC-Q, for MS/MS- and MS-based quantitation analysis. They are designed as generic platforms that can accommodate search results from SEQUEST and Mascot, as well as mzXML files converted from raw files produced by various mass spectrometers. Toward accurate quantitation analysis, Multi-Q determines detection limits of the user's instrument to filter out outliers and MaXIC-Q adopts stringent validation on our constructed projected ion mass spectra to ensure correct data for quantitation. PMID- 20013375 TI - Informatics and statistics for analyzing 2-d gel electrophoresis images. AB - Despite recent progress in "shotgun" peptide separation by integrated liquid chromatography and mass spectrometry (LC/MS), proteome coverage and reproducibility are still limited with this approach and obtaining enough replicate runs for biomarker discovery is a challenge. For these reasons, recent research demonstrates that there is a continuing need for protein separation by two-dimensional gel electrophoresis (2-DE). However, with traditional 2-DE informatics, the digitized images are reduced to symbolic data through spot detection and quantification before proteins are compared for differential expression by spot matching. Recently, a more robust and automated paradigm has emerged where gels are directly aligned in the image domain before spots are detected across the whole image set as a whole. In this chapter, we describe the methodology for both approaches and discuss the pitfalls present when reasoning statistically about the differential protein expression discovered. PMID- 20013377 TI - An overview of label-free quantitation methods in proteomics by mass spectrometry. AB - Protein quantification represents an important extension to identification proteomics, enabling the comparison of protein expression across different samples or treatments. Comparative protein quantification by mass spectrometry typically employs stable isotope incorporation, but recently, comparative quantification of label-free LC(n)-MS proteomics data has emerged as an alternative approach. In this chapter, we provide an overview of the different approaches for extracting quantitative data from label-free LC(n)-MS experiments. The computational procedure for recovering the quantitative information is outlined. Examples of statistical tests used to evaluate the relevance of results are also provided. PMID- 20013379 TI - Using the PRIDE proteomics identifications database for knowledge discovery and data analysis. AB - The PRIDE Proteomics Identifications Database provides users with the ability to explore and compare mass spectrometry-based proteomics experiments that reveal details of the protein expression found in a broad range of taxonomic groups, tissues and disease states. A PRIDE experiment typically includes identifications of proteins, peptides and protein modifications. Many of the submitted experiments also include processed peak lists representing the mass spectra that provide the evidence for these identifications.Since the inception of the PRIDE project, a number of tools supporting submission of data to PRIDE have been developed. Of particular note is the "PRIDE Converter" that has become the tool most frequently used for the production of PRIDE submissions at the time of writing.The PRIDE XML format has been expanded to provide submitters with the capacity to annotate fragment ion information on to peptide identifications and the fragmentation spectra that provide the experimental evidence for these peptides. A novel algorithm for annotating fragment ion information on to peptides and their evidential mass spectra has also been developed that will ultimately provide a route for evaluating the quality of peptide identifications arising from tandem mass spectrometry. This algorithm allows the visualisation of potential fragment ions on to the identified mass spectra, even where no such information has been submitted.In this chapter, we describe how PRIDE can be applied as a research tool and how the experiments in PRIDE can be compared and analysed. We also explore how complex queries can be constructed using the PRIDE BioMart. Finally, we will describe how the user can integrate PRIDE data with annotation from other resources, using federated BioMart queries. PMID- 20013378 TI - The PeptideAtlas Project. AB - PeptideAtlas is a multi-species compendium of peptides observed with tandem mass spectrometry methods. Raw mass spectrometer output files are collected from the community and reprocessed through a uniform analysis and validation pipeline that continues to advance. The results are loaded into a database and the information derived from the raw data is returned to the community via several web-based data exploration tools. The PeptideAtlas resource is useful for experiment planning, improving genome annotation, and other data mining projects. PeptideAtlas has become especially useful for planning targeted proteomics experiments. PMID- 20013380 TI - Molecular interactions and data standardisation. AB - Molecular interactions are crucial components of the cellular process. In order to understand this complex machinery, one needs to gather published data from various sources. Many projects have initiated the collection of interaction data for this purpose since 2002. However, the lack of standardisation previously made the task of aggregating datasets difficult. This issue has been resolved by the creation of Molecular Interaction standard in 2004 by members of the Proteomics Standards Initiative (PSI), a work group of the Human Proteome Organization (HUPO). Furthermore, major database providers have come together with the goal to exchange data in order to optimise laborious curation tasks. Finally, tools and frameworks have been created based on PSI-MI standards to facilitate the visualisation and analysis of molecular interaction data. PMID- 20013381 TI - Mass spectrometer output file format mzML. AB - Mass spectrometry is an important technique for analyzing proteins and other biomolecular compounds in biological samples. Each of the vendors of these mass spectrometers uses a different proprietary binary output file format, which has hindered data sharing and the development of open source software for downstream analysis. The solution has been to develop, with the full participation of academic researchers as well as software and hardware vendors, an open XML-based format for encoding mass spectrometer output files, and then to write software to use this format for archiving, sharing, and processing. This chapter presents the various components and information available for this format, mzML. In addition to the XML schema that defines the file structure, a controlled vocabulary provides clear terms and definitions for the spectral metadata, and a semantic validation rules mapping file allows the mzML semantic validator to insure that an mzML document complies with one of several levels of requirements. Complete documentation and example files insure that the format may be uniformly implemented. At the time of release, there already existed several implementations of the format and vendors have committed to supporting the format in their products. PMID- 20013382 TI - Managing experimental data using FuGE. AB - Data management and sharing in omics science is highly challenging due to the constant evolution of experimental techniques, the range of instrument types and software used for analysis, and the high volumes of data produced. The Functional Genomics Experiment (FuGE) Model was created to provide a model for capturing descriptions of sample processing, experimental protocols and multidimensional data for any kind of omics experiment. FuGE has two modes of action: (a) as a storage architecture for experimental workflows and (b) as a framework for building new technology-specific data standards.FuGE is an object model that is converted into an XML implementation for data exchange. Software toolkits have been developed for data handling and for bridging between XML data files and relational database implementations. FuGE has been adopted by the Proteomics Standards Initiative (PSI, http://www.psidev.info ) for building several new data formats, and it is being used in a variety of other experimental contexts, thus allowing data to be integrated across a range of experimental types to support Systems Biology approaches. This chapter provides a practical guide for laboratories or groups wishing to manage their data, and for developers wishing to create new data formats using FuGE. PMID- 20013383 TI - Proteomics data collection (ProDaC): publishing and collecting proteomics data sets in public repositories using standard formats. AB - In Proteomics, fast enhancements with regard to technology are responsible for the creation of huge data sets. Consequently, in 2006 the European Commission funded a Coordination Action named ProDaC (Proteomics Data Collection) within the 6th EU Framework Programme to foster a community-wide data collection and data sharing. The aims of ProDaC were the development of documentation and storage standards, setup of a standardized data submission pipeline and collection of data.To reach these goals, the necessary work was structured in six thematic fields (work packages): Standards for Proteomics Data Representation, Standards Implementation, Data Integration Tools, Proteomics Repository Adaptation, Data Flow Management, and Proteomics Data Exploitation. The methods building the basis of the respective fields and the achieved results are described in the following sections. PMID- 20013384 TI - Computational resources for the prediction and analysis of native disorder in proteins. AB - Proteomics attempts to characterise the gene products expressed in a cell or tissue via a range of biophysical techniques including crystallography and NMR and, more relevantly to this volume, chromatography and mass spectrometry. It is becoming increasingly clear that the native states of segments of many of the cellular proteins are not stable, folded structures, and much of the proteome is in an unfolded, disordered state. These proteins and their disordered segments have functionally interesting properties and provide novel challenges for the biophysical techniques that are used to study them. This chapter focuses on computational approaches to predicting such regions and analyzing the functions linked to them, and has implications for protein scientists who wish to study such properties as molecular recognition and post-translational modifications. We also discuss resources where the results of predictions have been collated, making them publicly available to the wider biological community. PMID- 20013385 TI - Utilization of liposomes for studying drug transfer and uptake. AB - On entry into the body of the patient, drugs have to overcome many barriers in order to reach the target. The knowledge of the ability of drugs to cross these barriers, which mostly consist of lipid membranes, is of utmost interest in pharmacy.High values of lipophilicity of a drug might be a good pre-requisite for crossing these barriers. It also led liposomologists to think that highly lipophilic drugs may "stick" in the lipophilic interior of liposomal phospholipid membranes and therefore these liposomes may act as a retard formulation of the lipophilic drug.The presented method here estimates the transfer time of lipophilic drugs between liposomal lipid bilayers. This may help to judge the presumed retardation function of a specific liposomal delivery system for a chosen lipophilic drug. PMID- 20013386 TI - The use of liposomes in the study of drug metabolism: a method to incorporate the enzymes of the cytochrome p450 monooxygenase system into phospholipid, bilayer vesicles. AB - Although lipids are essential for the optimal activity of the cytochromes P450 monooxygenase system, relatively little is known about the membrane environment in which these enzymes function. One approach used to mimic the structural arrangement of lipids and enzymes within the endoplasmic reticulum is to physically incorporate the cytochromes P450 and their redox partners in a vesicle bilayer of phospholipids. Several methods have been devised for this purpose. This chapter describes a method in which the P450 monooxygenase system is incorporated by first, solubilizing the enzymes and lipid with sodium glycocholate. After the protein and lipid aggregates are dispersed, the detergent is removed by adsorption using BioBeads SM-2 resin which leads to the formation of bilayer vesicles of phospholipid containing incorporated cytochrome P450 and NADPH cytochrome P450 reductase. This procedure requires relatively a short preparation time, provides concentrated reconstituted systems that can be used in a wide range of applications, allows for several enzyme samples to be prepared simultaneously so that different conditions can be compared, and results in minimal loss of active enzyme. PMID- 20013387 TI - Use of liposomes to study cellular osmosensors. AB - When cells are exposed to changes in the osmotic pressure of the external medium, they respond with mechanisms of osmoregulation. An increase of the extracellular osmolality leads to the accumulation of internal solutes by biosynthesis or uptake. Particular bacterial transporters act as osmosensors and respond to increased osmotic pressure by catalyzing uptake of compatible solutes. The functions of osmosensing, osmoregulation , and solute transport of these transporters can be analyzed in molecular detail after solubilization, isolation, and reconstitution into phospholipid vesicles. Using this approach, intrinsic functions of osmosensing transporters are studied in a defined hydrophilic (access to both sides of the membrane) and hydrophobic surrounding (phospholipid membrane), and free of putative interacting cofactors and regulatory proteins. PMID- 20013388 TI - Studying mechanosensitive ion channels using liposomes. AB - Mechanosensitive (MS) ion channels are the primary molecular transducers of mechanical force into electrical and/or chemical intracellular signals in living cells. They have been implicated in innumerable mechanosensory physiological processes including touch and pain sensation, hearing, blood pressure control, micturition, cell volume regulation, tissue growth, or cellular turgor control. Much of what we know about the basic physical principles underlying the conversion of mechanical force acting upon membranes of living cells into conformational changes of MS channels comes from studies of MS channels reconstituted into artificial liposomes. Using bacterial MS channels as a model, we have shown by reconstituting these channels into liposomes that there is a close relationship between the physico-chemical properties of the lipid bilayer and structural dynamics bringing about the function of these channels. PMID- 20013389 TI - Studying amino acid transport using liposomes. AB - The transport of amino acid across the membranes has great importance in cell metabolism. Specific experimental methodologies are required for measuring the vectorial reactions catalyzed by the membrane transporters. So far, the most widely used technique to study amino acid transport was the measure of amino acid flux in intact cell systems expressing a specific transporter. Some limitations in this procedure are caused by the presence of endogenous transporters and intracellular enzymes and by the inaccessibility of the intracellular compartment. Alternative experimental strategies which allow to reducing the interferences and improving the handling of the internal compartment would be useful to the amino acid transport knowledge.An experimental protocol, which makes use of liposomes to study the transport of amino acid mediated by the glutamine/amino acid (ASCT2) transporter, solubilized from rat kidney brush borders, is described. The procedure is based on the reconstitution of the transporter in liposomes by removal of detergent from mixed micelles of detergent, solubilized protein, and phospholipid. The transport is assayed in the formed proteoliposomes measuring the Na(+) dependent uptake of L: [(3)H]glutamine in antiport with internal L: -glutamine. This method allows measuring the transport activity under well controlled experimental conditions and permits performing experiments which cannot be realized in intact cell systems. PMID- 20013390 TI - Use of liposomes for studying interactions of soluble proteins with cellular membranes. AB - Methods are described that have been used for characterizing the interaction of the soluble bovine seminal plasma protein PDC-109 with liposomes. PDC-109 binds to bull sperm cells upon ejaculation and is an important modulating factor of sperm cell maturation. The binding of the protein to sperm cells is mediated via lipids of the sperm plasma membrane. Most of our current knowledge about the molecular mechanisms of PDC-109-membrane interaction has been obtained by studies employing lipid vesicles. The experimental strategy described here can be applied to investigate the interaction of soluble proteins with membranes in order to understand their physiological role. PMID- 20013392 TI - The reconstitution of actin polymerization on liposomes. AB - Membrane-associated actin polymerization is of considerable interest due to its role in cell migration and the motility of intracellular organelles. Intensive research efforts are underway to investigate the physiological role of membrane associated actin as well as the regulation and mechanics of actin assembly. Branched actin polymerization on membranes is catalyzed by the Arp2/3 complex. Signaling events leading to the activation of the guanosine triphosphate (GTP) binding protein Cdc42 stimulate Arp2/3-dependent actin polymerization. We have studied the role of Cdc42 at the Golgi apparatus in part by reconstituting actin polymerization on isolated Golgi membranes and on liposomes. In this manner, we showed that cytosolic proteins are sufficient for actin assembly on a phospholipid bilayer. Here we describe methods for the cell-free reconstitution of membrane-associated actin polymerization using liposomes and brain cytosol. PMID- 20013391 TI - Liposomal reconstitution of monotopic integral membrane proteins. AB - In spite of considerable progress in the methodology for reconstitution of membrane proteins into the liposomes, a successful reconstitution still appears to be more an art than a science. Reconstitution of membrane proteins into bilayers is required for establishing several aspects of the functions of membrane proteins and lipids and for elaborating models of naturally occurring membranes.Cyclooxygenase (COX)-1 and -2 (also prostaglandin endoperoxide H(2) synthase, PGHS-1 and -2) belong to the class of monotopic membrane proteins. Membrane-binding domains of both COX-1 and -2 contain four short, consecutive, amphipathic alpha-helices (A, B, C, and D). Crystal structures of the COXs indicate that basic, hydrophobic, and aromatic residues in the membrane-binding domain are oriented away from the protein core and form a surface on the enzyme, which has been proposed to interact with the lipid bilayer (1).In this chapter, we describe a fast and efficient method for direct incorporation of COX-1 and -2 isozymes - as models for monotopic integral membrane proteins - into preformed liposomes containing fatty acids without loss of activity. PMID- 20013393 TI - Electroformation of giant unilamellar vesicles from native membranes and organic lipid mixtures for the study of lipid domains under physiological ionic-strength conditions. AB - Giant unilamellar vesicles (GUVs) constitute a cell-sized model membrane system that allows direct visualization of particular membrane-related phenomena, such as domain formation, at the level of single vesicles using fluorescence microscopy-related techniques. Currently available protocols for the preparation of GUVs work only at very low salt concentrations, thus precluding experimentation under physiological conditions. In addition, the GUVs thus obtained lack membrane compositional asymmetry. Here we show how to prepare GUVs using a new protocol based on the electroformation method either from native membranes or organic lipid mixtures at physiological ionic strength. Additionally, we describe methods to test whether membrane proteins and glycosphingolipids preserve their natural orientation after electroformation of GUVs composed of native membranes. PMID- 20013394 TI - Visualization of lipid domain-specific protein sorting in giant unilamellar vesicles. AB - Recent studies suggest that phospholipids in the plasma membrane of mammalian cells are not homogenously distributed but may form domains either by lipid-lipid interactions or/and as consequence of lipid-protein interactions. Such lipid compartments may act as protein recruiting platforms which, for example, are essential components of cell signaling pathways. Model membrane systems with a defined lipid composition are ideally suited to study domain-specific interactions of peripheral and integral membrane proteins. Giant unilamellar vesicles (GUVs) offer the opportunity to directly visualize in parallel, both the lateral lipid domains and the interaction sites of proteins using fluorescence microscopy. The application of GUVs is exemplarly illustrated for studying domain specific interactions of the protein alpha-synuclein and the domain-specific distribution of synthetic transmembrane peptides. PMID- 20013395 TI - Biosynthesis of proteins inside liposomes. AB - Protein expression is the most complex metabolic reaction that has been encapsulated in liposomes, mainly as an intermediate step toward the synthesis of minimal semisynthetic cells. Although there are different experimental approaches to achieving the synthesis of proteins inside liposomes and it is therefore not possible to give a standard recipe, all methods follow a general strategy, which is briefly discussed. On this basis, we provide general indications for designing and realizing protein-expressing liposomes. Our approach for the green fluorescent protein expression inside 200-nm extruded vesicles is described in detail. PMID- 20013397 TI - Use of liposomes to evaluate the role of membrane interactions on antioxidant activity. AB - Cellular membranes, which contain abundant phospholipids, such as phosphatidylcholine, are major targets subjected to the damage caused by free radicals. Cellular damage due to lipid oxidation is strongly associated with ageing, carcinogenesis and other diseases. In addition, lipid oxidation is an important deteriorative reaction in the processing and storage of lipid containing foods. Liposomes have been used extensively as biological models for in vitro lipid oxidation studies. The resemblance between the liposomal and membrane bilayer core makes liposomes a very useful tool to investigate the significance of the antioxidant-membrane interactions for antioxidant activity. The antioxidant activity of a compound is strongly influenced by numerous factors including the nature of the lipid substrate, the hydrophilic-lipophilic balance of the antioxidant, the physical and chemical environments of the lipids, and various other interfacial interactions. Thus, compounds that are effective antioxidants in one model system or food matrix may be unsuitable in other systems.This chapter describes fluorescent probes-based methods commonly used for testing antioxidant activity in liposomes and stresses the need to combine antioxidant assays and drug-membrane interaction studies to get a better description of the antioxidants' profile considering their location in lipid bilayer and their effect on membrane fluidity and consequently provide additional information to that obtained currently from assays performed in aqueous buffer media. PMID- 20013396 TI - Study of respiratory cytochromes in liposomes. AB - Important findings regarding the structure and function of respiratory cytochromes have been made from the study of these hemeproteins associated to liposomes. These studies contributed to the comprehension of the biological role of these proteins in the electron transfer process, the regulatory mechanisms, the energy transduction mechanisms, the protein sites that interact with mitochondrial membranes and the role played by the non-redox subunits present in the protein complexes of the respiratory chain of eukaryotes. In this chapter, the protocols developed to study cytochrome bc (1) activity in liposomes and the binding of cytochrome c to lipid bilayers is presented . The former protocol was developed to study the mechanism of energy transduction related to the topology of the components of bc (1) complex in the mitochondrial membrane. These studies were done with purified cytochrome bc (1) complexes reconstituted into potassium loaded vesicles. The latter protocol was developed to study the influence of pH, DeltapH, and DeltaPsi on the interaction of cytochrome c with liposomes that mimic the inner mitochondrial membrane. PMID- 20013398 TI - Studying colloidal aggregation using liposomes. AB - Colloidal aggregation using liposomes has been studied in this chapter. As criteria of stability, the stability factor, an extension of the DLVO theory of colloidal stability, the fractal dimension of the liposome aggregates and the different regimes of aggregation (RLCA and DLCA) and the temperature have been used. PMID- 20013399 TI - Assessment of liposome-cell interactions. AB - Liposome-cell interactions have been assessed for over 30 years now by an enormous variety of approaches and methods. In-depth knowledge of liposome-cell interaction is still very relevant since new concepts and applications applying liposomes are being developed every day. This chapter does not aim to give a complete overview on methods to assess liposome-cell interactions but merely gives a handle to approach the assessment of liposome-cell interaction by describing some well-established methods that also allow for modification to adapt them for your specific research questions. PMID- 20013400 TI - Methods to monitor liposome fusion, permeability, and interaction with cells. AB - We describe fluorescence assays for membrane fusion involving the fusion of liposomes with each other and with cultured cells, fluorescence methods to assess liposome uptake by cells and the intracellular delivery of liposome contents, and assays to evaluate liposome membrane permeability. The Tb/DPA and ANTS/DPX assays monitor the intermixing of aqueous contents of liposomes. The NBD-PE/Rhodamine-PE assay follows the intermixing of liposomal lipids. A variation of this method is suitable for detecting the mixing of the inner monolayers of liposomes. The lipid mixing assay is also used to study the fusion of cationic liposomes and lipoplexes with cultured cells. The intracellular delivery of liposome contents are monitored, via fluorescence microscopy or flow cytometry, by measuring the release of calcein from the liposome interior, and normalized to cell-associated liposomes quantitated with Rhodamine-PE in the membrane of the same liposomes. The release of liposome contents is monitored by the increase in fluorescence of encapsulated carboxyfluorescein, calcein, or ANTS/DPX, or by the decrease in fluorescence of encapsulated Tb/DPA. PMID- 20013401 TI - The use of isothermal titration calorimetry to study multidrug transport proteins in liposomes. AB - Biophysical measurements of multidrug transporters in vitro can often be of limited relevance to the natural in vivo behavior. In particular, the properties of transporters when removed from their native bilayer and solubilized in detergents or lipids can differ significantly from their in vivo properties, reducing the value of in vitro measurements for the design of antagonists to the transporters. This problem can be addressed by studying the transport protein in liposomes in which the properties of the liposome bilayer are altered through systematic changes in lipid composition. Isothermal titration calorimetry can be used to determine the properties of the lipid-reconstituted protein in bilayers of different lipid compositions as well as to quantify the percentage recovery of functional protein in different lipids. Both these measurements lead to an accurate analysis of substrate binding activity. The approach is illustrated here for the small multidrug transport protein, EmrE from Escherichia coli. The percentage of functional EmrE successfully reconstituted into liposome depends on lipid composition. Differences in ligand binding and subtle differences in the secondary structure also occur in different lipid compositions. PMID- 20013403 TI - Membrane translocation assayed by fluorescence spectroscopy. AB - Assessing the ability of biomolecules or drugs to overcome lipid membranes in a receptor-independent way is of great importance in both basic research and applications involving the use of liposomes. A combination of uptake, release, and dilution experiments performed by steady-state fluorescence spectroscopy provides a powerful, straightforward, and inexpensive way of monitoring membrane translocation of fluorescent compounds. This is particularly true for peptides and proteins carrying intrinsic tryptophan residues, which eliminates the need for attaching extrinsic labeling moieties to the compound of interest. The approach encompasses three different kinds of fluorescence titrations and some simple calculations that can be carried out in a spreadsheet program. A complete set of experiments and data analyses can typically be completed within two days. PMID- 20013402 TI - Studying lipid organization in biological membranes using liposomes and EPR spin labeling. AB - Electron paramagnetic resonance (EPR) spin-labeling methods provide a unique opportunity to determine the lateral organization of lipid bilayer membranes by discrimination of coexisting membrane domains or coexisting membrane phases. In some cases, coexisting membrane domains can be characterized without the need for their physical separation by profiles of alkyl chain order, fluidity, hydrophobicity, and oxygen diffusion-concentration product in situ. This chapter briefly explains how EPR spin-labeling methods can be used to obtain the above mentioned profiles across lipid bilayer membranes (liposomes). These procedures will be illustrated by EPR measurements performed on multilamellar liposomes made of lipid extracts from cortical and nuclear fractions of the fiber cell plasma membrane of a cow-eye lens. To better elucidate the major factors that determine membrane properties, results for eye lens lipid membranes and simple model membranes that resemble the basic lipid composition of biological membranes will be compared. PMID- 20013404 TI - Interaction of lipids and ligands with nicotinic acetylcholine receptor vesicles assessed by electron paramagnetic resonance spectroscopy. AB - Electron paramagnetic resonance (EPR) spectroscopy is a powerful technique that permits the study of membrane-embedded proteins in its lipid environment by assessing the interaction of spin labels with the protein in its natural environment (i.e., native membranes) or in reconstituted systems prepared with exogenous lipid species. Nicotinic acetylcholine receptors (AChRs) contain a large surface in intimate contact with the lipid membrane. AChRs, members of the Cys-loop receptor superfamily, have essential functional roles in the nervous system and its malfunctioning has been considered as the origin of several neurological diseases including Alzheimer's disease, drug addiction, depression, and schizophrenia. In this regard, these receptors have been extensively studied as therapeutic targets for the action of several drugs. The majority of the marketed medications bind to the neurotransmitter sites, the so-called agonists. However, several drugs, some of them still in clinical trials, interact with non competitive antagonist (NCA) binding sites. A potential location for these binding sites is the proper ion channel, blocking ion flux and thus, inhibiting membrane depolarization. However, several NCAs also bind to the lipid-protein interface, modulating the AChR functional properties. The best known examples of these NCAs are local and general anesthetics. Several endogenous molecules such as free fatty acids and neurosteroids also bind to the lipid-protein interface, probably mediating important physiological functions. Phospholipids, natural components of lipid membranes interacting with the AChR, are also essential to maintain the structural and functional properties of the AChR. EPR studies showed that local anesthetics bind to the lipid-protein interface by essentially the same dynamic mechanisms found in lipids, and that local and general anesthetics preferably decrease the phospholipid but not the fatty acid interactions with the AChR. This is consistent with the existence of annular and non-annular lipid domains on the AChR. PMID- 20013405 TI - Environmental scanning electron microscope imaging of vesicle systems. AB - The structural characteristics of liposomes have been widely investigated and there is certainly a strong understanding of their morphological characteristics. Imaging of these systems, using techniques such as freeze-fracturing methods, transmission electron microscopy, and cryo-electron imaging, has allowed us to appreciate their bilayer structures and factors that influence this. However, there are a few methods that study these systems in their natural hydrated state; commonly, the liposomes are visualized after drying, staining and/or fixation of the vesicles. Environmental scanning electron microscopy (ESEM) offers the ability to image a liposome in its hydrated state without the need for prior sample preparation. We were the first to use ESEM to study the liposomes and niosomes, and have been able to dynamically follow the hydration of lipid films and changes in liposome suspensions as water condenses onto, or evaporates from, the sample in real-time. This provides an insight into the resistance of liposomes to coalescence during dehydration, thereby providing an alternative assay for liposome formulation and stability. PMID- 20013406 TI - Freeze-fracture electron microscopy on domains in lipid mono- and bilayer on nano resolution scale. AB - Freeze-fracture electron microscopy (FFEM) as a cryo-fixation, replica, and transmission electron microscopy technique is unique in membrane bilayer and lipid monolayer research because it enables us, to excess and visualize pattern such as domains in the hydrophobic center of lipid bilayer as well as the lipid/gas interface of the lipid monolayer. Since one of the preparatory steps of this technique includes fracturing the frozen sample and, since during this fracturing process the fracture plane follows the area of weakest forces, these areas are exposed allowing us to explore the pattern built up by lipids and/or intrinsic proteins and which are also initiated by peptides, drugs, and toxins reaching into these normally hard to access areas. Furthermore, FFEM as a replica technique is applicable to objects of a large size range and combines detailed imaging of fine structures down to nano-resolution scale within images of larger biological or artificial objects up to several ten's of micrometers in size.Biological membranes consist of a multitude of components which can self organize into rafts or domains within the fluid bilayer characterized by lateral inhomogeneities in chemical composition and/or physical properties. These domains seem to play important roles in signal transduction and membrane traffic. Furthermore, lipid domains are important in health and disease and make an interesting target for pharmacological approaches in cure and prevention of diseases such as Alzheimer, Parkinson, cardiovascular and prion diseases, systemic lupus erythematosus and HIV. As a cryofixation technique FFEM is a very powerful tool to capture such domains in a probe-free mode and explore their dynamics on a nano-resolution scale. PMID- 20013407 TI - Atomic force microscopy for the characterization of proteoliposomes. AB - Atomic Force Microscopy (AFM) is a useful tool for the visualization of soft biological samples in a nanoscale resolution. In the study presented here, the surface morphology ofP-selectin and Transferrin modified proteoliposomes were investigated in air and under water. The proteins were visualized without pre functionalization or staining. PMID- 20013408 TI - Method of simultaneous analysis of liposome components using HPTLC/FID. AB - Liposomes are composed of different kind of lipids or lipophilic substances and are used as carriers of bioactive molecules. The characterization of the prepared liposomes consists of the calculation of the drug to lipid molar ratio by measuring the lipids and the encapsulated molecule.The present work describes an analytical methodology on simultaneous determination of all the lipid ingredients of the liposome formulation, using Thin Layer Chromatography coupled with a Flame Ionization Detector (TLC/FID), using the least possible sample quantity. The method consists of a chromatographic separation of the liposomal ingredients on silica gel scintillated on quartz rods and subsequent detection of the ingredients by scanning the rods by a hydrogen flame. The produced ions are detected by a Flame Ionization Detector and the signal is converted to a chromatogram.This method may be applied on every step of the liposome preparation for examining the quality of the raw materials, tracking possible errors of the preparation procedure and finally analyzing the content of the final liposomal composition. PMID- 20013409 TI - Viscometric analysis of DNA-lipid complexes. AB - DNA-cationic lipid complexes, "lipoplexes", are used as gene carriers for molecular biology and gene therapy applications. Colloidal properties of lipoplexes can be determined by viscometric analysis. (1) The shape parameter of lipoplexes can be one of the factors affecting transfection efficiency; (2) the volume fraction of free liposomes remaining after lipoplex formation can be used as an index of purity of the lipoplex product; (3) the shear dependence of the viscosity of a diluted lipoplex suspension can be used as a macroscopic shape factor: (4) the attraction force parameter between particles can be a colloidal stability factor. These properties should be characterized and specified for process control of lipoplex production and quality control of lipoplex products.We describe an automated mini-capillary viscometer for a sample volume of 0.5 ml, and its application to the characterizations of lipoplexes. We show a procedure for viscosity measurements and provide a calculation using complexes of plant DNA-distearyldimethylammonium chloride (DDAC) at a charged ratio of 1:4 ( /+), in which the amount of DNA is less than 250 microg. The prolate/ellipsoidal axial ratio, a/b, was found to be 70. Determination of the shape parameter with a/b is found to be better than that with other shape parameters, e.g., alpha of the Sakurada equation, because fractionation of the particle size is not necessary. By the proposed method, colloidal parameters of lipoplexes and bioactive polymer complexes are characterized quantitatively. PMID- 20013410 TI - Fluorometric Analysis of Individual Cationic Lipid-DNA Complexes. AB - Lipoplex preparations are heterogeneous mixtures of lipoplex particles of different structures. As these structures determine the efficiency of the delivery of genetic material, it is important to characterize the distribution of particles of different types in lipoplex preparations with good statistics. We describe the application of flow fluorometry which allows producing such distributions (in terms of lipoplex particle size and composition) within minutes using basic flow cytometer (Anal Biochem 341:230-240, 2005). PMID- 20013411 TI - Fluorescence resonance energy transfer-based analysis of lipoplexes. AB - We describe three applications of the FRET technique to analysis of structural and thermodynamic properties of cationic lipids and their complexes with DNA. (1) Lipid mixing assay to determine the degree to which individual vesicles undergo fusion upon complex formation. (2) DNA binding assay to obtain cationic lipid-DNA binding curves for thermodynamic analysis or binding stoichiometry characterization. (3) DNA spacing assay to determine changes in DNA packing in the multilamellar lipoplex particle. PMID- 20013412 TI - Analysis of lipoplex structure and lipid phase changes. AB - Efficient delivery of genetic material to cells is needed for tasks of utmost importance in the laboratory and clinic, such as gene transfection and gene silencing. Synthetic cationic lipids can be used as delivery vehicles for nucleic acids and are now considered the most promising nonviral gene carriers. They form complexes (lipoplexes) with the polyanionic nucleic acids. A critical obstacle for clinical application of the lipid-mediated DNA delivery (lipofection) is its unsatisfactory efficiency for many cell types. Understanding the mechanism of lipid-mediated DNA delivery is essential for their successful application, as well as for a rational design and synthesis of novel cationic lipoid compounds for enhanced gene delivery. A viewpoint now emerging is that the critical factor in lipid-mediated transfection is the structural evolution of lipoplexes within the cell, upon interacting and mixing with cellular lipids. In particular, recent studies showed that the phase evolution of lipoplex lipids upon interaction and mixing with membrane lipids appears to be decisive for transfection success: specifically, lamellar lipoplex formulations, which were readily susceptible to undergoing lamellar-nonlamellar phase transition upon mixing with cellular lipids and were found rather consistently associated with superior transfection potency, presumably as a result of facilitated DNA release. Thus, understanding the lipoplex structure and the phase changes upon interacting with membrane lipids is important for the successful application of the cationic lipids as gene carriers. PMID- 20013413 TI - Fluorescence methods for evaluating lipoplex-mediated gene delivery. AB - The biological activity of cationic liposome/DNA complexes ("lipoplexes") is strongly dependent on their ability to protect DNA and to interact with cells, including binding to the cell surface, internalization via endocytosis and cytoplasmic delivery of the DNA. In this chapter, we describe a number of methods and procedures to study these processes, based on the use of fluorescent probes. PMID- 20013415 TI - Spectral bio-imaging and confocal imaging of the intracellular distribution of lipoplexes. AB - The intracellular distribution of nanoparticular drug delivery systems is very complex, but its investigation yields high potential for further development and optimization of these systems.In the following chapter, we introduce the application of fluorescent imaging techniques in order to highlight uptake and cellular processing of nanoparticular drug delivery systems (e.g., liposomal drug delivery systems). We selected a combination of different protocols for the staining of the most important endocytic compartments and organelles. The presented imaging systems are appropriate to detect liposomal drug delivery systems localized in these cellular structures. PMID- 20013414 TI - FRET imaging of cells transfected with siRNA/liposome complexes. AB - By monitoring the efficiency of fluorescence resonance energy transfer of dyes attached to the different strands of siRNA, the structural integrity of the latter can be traced inside cells. Here, the experimental details of dye-labeled siRNA construction, tissue culture, and transfection with liposomally formulated siRNAs are given, as well as the conditions for confocal microscopy and an algorithm allowing the visualization of intact siRNA after image data treatment. The method allows rapid screening of different liposomal siRNA formulations, obtained by small scale dual asymmetric centrifugation with high entrapping efficiency. PMID- 20013416 TI - Techniques for loading technetium-99m and rhenium-186/188 radionuclides into pre formed liposomes for diagnostic imaging and radionuclide therapy. AB - Liposomes can serve as carriers of radionuclides for diagnostic imaging and therapeutic applications. Herein, procedures are outlined for radiolabeling liposomes with the gamma-emitting radionuclide, technetium-99m ((99m)Tc), for non invasive detection of disease and for monitoring the pharmacokinetics and biodistribution of liposomal drugs, and/or with therapeutic beta-emitting radionuclides, rhenium-186/188 ((186/188)Re), for radionuclide therapy. These efficient and practical liposome radiolabeling methods use a post-labeling mechanism to load (99m)Tc or (186/188)Re into pre-formed liposomes prepared in advance of the labeling procedure. For all liposome radiolabeling methods described, a lipophilic chelator is used to transport (99m)Tc or (186/188)Re across the lipid bilayer of the pre-formed liposomes. Once within the liposome interior, the pre-encapsulated glutathione or ammonium sulfate (pH) gradient provides for stable entrapment of the (99m)Tc and (186/188)Re within the liposomes. In the first method, (99m)Tc is transported across the lipid bilayer by the lipophilic chelator, hexamethylpropyleneamine oxime (HMPAO) and (99m)Tc HMPAO becomes trapped by interaction with the pre-encapsulated glutathione within the liposomes. In the second method, (99m)Tc or (186/188)Re is transported across the lipid bilayer by the lipophilic chelator, N,N-bis(2-mercaptoethyl)-N',N' diethylethylenediamine (BMEDA), and (99m)Tc-BMEDA or (186/188)Re-BMEDA becomes trapped by interaction with pre-encapsulated glutathione within the liposomes. In the third method, an ammonium sulfate (pH) gradient loading technique is employed using liposomes with an extraliposomal pH of 7.4 and an interior pH of 5.1. BMEDA, which is lipophilic at pH 7.4, serves as a lipophilic chelator for (99m)Tc or (186/188)Re to transport the radionuclides across the lipid bilayer. Once within the more acidic liposome interior, (99m)Tc/(186/188)Re-BMEDA complex becomes protonated and more hydrophilic, which results in stable entrapment of the (99m)Tc/(186/188)Re-BMEDA complex within the liposomes. Since many commercially available liposomal drugs use an ammonium sulfate (pH) gradient for drug loading, these liposomal drugs can be directly radiolabeled with (99m)Tc BMEDA for non-invasive monitoring of tissue distribution during treatment or with (186/188)Re-BMEDA for combination chemo-radionuclide therapy. PMID- 20013417 TI - Fluorescence correlation spectroscopy for the study of membrane dynamics and organization in giant unilamellar vesicles. AB - Fluorescence correlation spectroscopy (FCS) is a powerful technique to study the lateral organization of membranes. It measures fluorescence intensity fluctuations in the single molecule regime and allows the determination of diffusion coefficients. When applied to lipid membranes, their fluidity and lipid phase can be estimated from the diffusion rates of fluorescent particles partitioned to the membrane. Here, we describe the theoretical basis of FCS and discuss the z-scan approach for measurements on lipid membranes. We also list the materials necessary for a FCS experiment on giant unilamellar vesicles (GUVs). Finally, we present simple protocols for the preparation of GUVs and the acquisition and analysis of FCS data on the vesicles, so that diffusion coefficients of fluorescent probes within lipid membranes can be estimated. PMID- 20013418 TI - Liposome biodistribution via europium complexes. AB - The drug-delivery field needs tools to follow vector biodistribution. Radioactive tracers and conventional fluorophores are widely used. We propose here to use europium complexes. Use of pulsed light source time-resolved fluorimetry takes into account the fluorescence decay time of the lanthanide chelates to gain sensitivity in biological media. The method was developed to follow liposome biodistribution. Octadecyl-DTPA.Eu compound has been prepared and incorporated into liposomes without alteration of its fluorescence signal. The method has been validated by comparison with fluorophore-labelled liposomes. The way to proceed to use this method for liposomes or other vectors is detailed. PMID- 20013419 TI - Biosensor-based evaluation of liposomal binding behavior. AB - Biosensors can be regarded as analytical devices that transform biologically given facts, such as the appearance of physiological substrates, or biological recognition processes of ligands and receptors into detectable signals without the need of further labeling. This chapter introduces acoustic wave sensors as mass-sensitive tools to investigate the liposomal binding behavior onto simulated biological surfaces. These sensors do not only allow for quantification of the liposomal binding intensity, but further analytical readings give insight into the liposomal appearance at the binding site, e.g., deformation or fusion. Since the liposomal behavior at the target binding site might have strong impact on therapeutic effects, a prediction of liposomal appearance and a controlled modulation thereof appear possible with the help of biosensors.Here, the function of a quartz crystal microbalance (QCM) and the bio-functionalization of quartz sensors are reported for a series of liposomal binding experiments. Liposomes containing biotin as model ligands were selected to evaluate their binding to avidin-modified sensors. The data, representing binding intensity and liposome deformation, are explained with respect to the role of binding strength and lipid composition for liposomal behavior. PMID- 20013420 TI - Use of liposomes to study vesicular transport. AB - Liposomes have been utilized for variety of membrane transport studies including clathrin-mediated endocytosis. Here we introduce clathrin-coated structures that are generated on large unilamellar liposomes by incubating with clathrin coat proteins. Large unilamellar liposomes are also used to reconstitute vesicle formation in a cell-free system, and the vesicle formation can be quantified by using dynamic light scattering (DLS). Furthermore, phagocytosis assay using liposome-conjugated styrene beads is demonstrated. PMID- 20013421 TI - [Turkish Journal of Psychiatry]. PMID- 20013422 TI - [Predictors of social functioning in schizophrenia]. AB - OBJECTIVE: Impaired social functioning is a characteristic feature of schizophrenia. The aim of this study was to determine the factors that predict social functioning in schizophrenia. METHOD: The study included 100 patients that met DSM-IV diagnostic criteria for schizophrenia. All the patients were evaluated using the Scale for the Assessment of Negative Symptoms, Scale for the Assessment of Positive Symptoms, Calgary Depression Scale for Schizophrenia, Facial Emotion Identification Test, Facial Emotion Discrimination Test, Digit Span Test, Rey's Auditory Verbal Learning Test, Trail Making Test, and Social Functioning Scale. Marital status, living arrangements, and working status of the patients were also recorded. Pearson's correlation analysis and multiple stepwise regression analysis were used to determine the relationships between the research variables and social functioning. Categorical variables were compared with the independent samples t test. RESULTS: Negative symptoms, level of education, and facial emotion identification were significant predictors of social functioning total score. Negative symptoms were the most important predictor of almost all domains of social functioning. Age and duration of illness were not associated with social functioning. There were no gender differences in terms of social functioning. Married patients had higher social functioning scores. Employed patients had lower negative symptom scores and higher social functioning scores. CONCLUSION: Negative symptoms were the most important predictors of social functioning in our study. PMID- 20013423 TI - [Assessment of executive functions in social phobia patients using the Wisconsin Card Sorting Test]. AB - OBJECTIVE: Problem solving and adjusting responses according to feedback are among the executive functions that may be impaired in social phobia patients. The objective of this study was to compare social phobia patients' Wisconsin Card Sorting Test scores with those of controls; thus, our aim was to examine executive functions in social phobia patients. METHOD: The study included 36 social phobia patients (16 female [44.4%] and 20 male [55.6%]) whose age, sex, and level of education were matched with those of a healthy control group. Participants were administered the Structured Clinical Interview for DSM-IV Axis I Disorders, Beck Depression Inventory, State-Trait Anxiety Inventory, Liebowitz Social Anxiety Scale along, and Wisconsin Card Sorting Test. RESULTS: Patients with social phobia scored lower than the control group in terms of the total number of correct responses, number of categories completed, and percentage of conceptual level responses on the Wisconsin Card Sorting Test. The total number of errors and total non-preservative errors were elevated in the patient group. No differences were observed in preservative errors and set-maintenance between the patient and control groups. The number of correct responses and the percentage of conceptual level responses were negatively correlated with trait anxiety and social avoidance scores, whereas the number of errors was positively correlated with trait anxiety and social avoidance scores. Non-preservative errors were positively correlated with state-anxiety, social fear, and social avoidance scores. CONCLUSION: Working memory in the social phobia patients was impaired, as compared to that of the healthy controls. High social anxiety scores had a negative impact on working memory. PMID- 20013424 TI - [What does the subjective assessment of cognitive functioning measure in bipolar disorder? Correlation with the objective assessment of cognitive functioning]. AB - OBJECTIVE: Even in remission, cognitive deficits are frequently observed in bipolar disorder. Because these deficits are assessed with complex and comprehensive tests, they seem to be neglected in routine daily practice; however, assessing them with short self-rated questionnaires could be beneficial. This study aimed to investigate the correlation between objective and subjective cognitive assessment in bipolar disorder. METHOD: The study included 38 patients with bipolar disorder that were in remission for at least 6 months and 19 healthy controls. The Stroop Test and Auditory Verbal Learning Test were used for the objective assessment of cognitive functioning, and the cognitive functioning subscale of the Bipolar Disorder Functioning Questionnaire was used for the subjective assessment of cognitive functioning. Mood symptoms were assessed using the Hamilton Depression Rating Scale and Young Mania Rating Scale. RESULTS: Based on objective assessment there were significant differences in all domains of the Auditory Verbal Learning Test between the bipolar patients and healthy controls, whereas there weren't any significant difference in Stroop Test results. Based on subjective assessment there were significant differences between the bipolar patients and healthy controls in terms of cognitive functioning subscale total score and first item score, but not in terms of the other items. When the correlation between the objective and subjective cognitive assessments was investigated, total score of the cognitive functioning subscale and 2 items--1. to run simple calculations in one's mind and 2. gathering and performing what one is asked to do--were moderately to weakly correlated with immediate recall, verbal learning, recognition, retrieval, and reading shape color. No correlation was observed between the other items of the cognitive functioning subscale. CONCLUSION: Based on subjective assessment of cognitive functioning, the bipolar patients tended to over-estimate themselves. Furthermore, bipolar patients tended to report more deficits correlated with objective cognitive assessment when they had to display active performance, whereas they did not report cognitive deficits at this level when such a condition was not present. PMID- 20013425 TI - [Confirmatory factor analysis of schizotypal personality traits in university students]. AB - OBJECTIVE: Investigating the factor structure of schizotypal traits in normal population is important to describe the clinical phenotypes associated with susceptibility to schizophrenia. Schizotypal Personality Questionnaire (SPQ) is a commonly used self-rated measure to assess schizotypal traits. While Raine's three-factorial model is most commonly supported model explaining factor structure of schizotypal traits, there is also evidence supporting alternative models. The aim of this study is compare the goodness-of-fitness of various models about factor structure of the SPQ in a substantial number of university students. METHOD: 1059 university students were participated in the study. Alternative models regarding factor structure of the SPQ were compared with confirmatory factor analysis (CFA). The effect of gender on the factor structure of schizotypal traits is also studied. RESULTS: The 4 factorial model that included cognitive-perceptual, paranoid, interpersonal and disorganized dimensions fit the data the best. Raine's three factorial model did not fit the data adequately. However, after minor modifications, Raine's model also had a satisfactory goodness-of-fit. Gender had no effect on the factor structure of the SPQ. CONCLUSION: Results of these study supported 4-factorial model of Stefanis and modified version of Raine's model to explain factor structure of schizotypal traits. The structure of schizotypal traits is in parallel with structure of symptom dimensions in schizophrenia. This outcome is compatible with the views seeing schizotypal traits in general population in continuum with schizophrenia spectrum disorders. PMID- 20013426 TI - [Differences in cerebral blood flow following risperidone treatment in children with autistic disorder]. AB - OBJECTIVE: Functional changes in the brains of autistic children due to risperidone treatment and theirs relationship to the symptom clusters are yet unknown. In this autistic disorder case series we aimed to comparatively evaluate the clinical findings before and after risperidone treatment, and regional cerebral blood flow (rCBF) findings with 99mTc-hexamethylpropyleneamine oxime (HMPAO) brain SPECT. METHOD: Eleven autistic patients (age range: 6-7 years; 4 girls, 7 boys) received risperidone therapy (1.5-2.5 mg d(-1)) and were followed up for 3 months. All the patients underwent neurologic examinations, psychometric examinations, and SPECT imaging, both at the start of risperidone treatment and 3 months after the treatment started. Clinical observations, and the observations of parents and teachers were recorded. These results were compared with cerebral perfusion indices obtained from SPECT data. RESULTS: After 3 months of treatment changes in rCBF were observed in various regions and to varying degrees. We observed relationships between clinical symptoms and pre-therapy rCBF findings, and between clinical improvement and rCBF changes. CONCLUSION: Findings in the present case series are the first to demonstrate a relationship between clinical improvement and regional perfusion patterns after risperidone treatment. We think that these findings may contribute to the understanding of the neurofunctional mechanisms and hypothetical models of autism. PMID- 20013427 TI - [The effect of group-based psychodrama therapy on decreasing the level of aggression in adolescents]. AB - PURPOSE: This study aimed to examine the effect of group-based psychodrama therapy on the level aggression in adolescents. METHOD: The study included 23 students from Nezihe Yalvac Anatolian Vocational High School of Hotel Management and Tourism that had high aggression scores. Eleven of the participants (6 female, 5 male) constituted the experimental group and 12 (6 male, 6 female) were in the control group. The 34-item Aggression Scale was used to measure level of aggression. We utilized mixed pattern design including experiment-control, pre test and post test and follow up. The experimental group participated in group based psychodrama therapy once a week for 90 minutes, for 14 weeks in total. The Aggression Scale was administered to the experimental and control groups before and after treatment; it was additionally administered to the experimental group 16 weeks after treatment. Data were analyzed using ANCOVA and dependent samples t tests. RESULTS: Our analysis shows that group-based psychodrama had an effect on the experimental group in terms of total aggression, anger, hostility, and indirect aggression scores (F=65.109, F=20.175, F=18.593, F=40.987, respectively, P<.001). There was no effect of the group-based treatment on verbal or physical aggression scores. Follow-up indicated that the effect of the therapy was still measureable 16 weeks after the cessation of the therapy. DISCUSSION: Results of the present study indicate that group-based psychodrama therapy decreased the level of aggression in the experimental group. Current findings are discussed with reference to the literature. Recommendations for further research and for psychiatric counselors are provided. PMID- 20013428 TI - [Psychometric properties of the Turkish Working Alliance Inventory-Observer Form]. AB - OBJECTIVE: This study aimed to examine the reliability and validity of the Turkish version of the Working Alliance Inventory-Observer Form (WAI-O) (Titchenor and Hill, 1989). METHOD: The study included 18 videotape records of 6 therapists conducting short-term cognitive behavioral therapy. The videotape records were chosen randomly from the pool of Hacettepe University Psychotherapy Research Laboratory (HU-PRL) and coded by 3 judges. Intra-class correlation (ICC) coefficients for the scale were calculated using the average measure method to determine the inter judge reliability coefficients for the data obtained from the 3 judges. RESULTS: The ICC coefficients were 0.73, 0.70, and 0.78 for the bond, goal, and task dimensions, respectively. The ICC coefficient was 0.75 for the inventory total score. The internal validity of the scale was alpha=0.90. The item total correlation ranged between 0.84 and 0.96. DISCUSSION: On a preliminary basis our findings indicate that the Turkish version of the WAI-O Short Form has acceptable levels of reliability and validity for clinical and research use in Turkey. PMID- 20013429 TI - [Effects of valproate on male reproductive functions]. AB - Valproate is an antiepileptic commonly used in the treatment of psychiatric and neurologic diseases. Some of the most frequently seen side effects affect the gastrointestinal, neurological, and hematological systems. Its side effects on reproductive functions have not been sufficiently studied. The use of valproate by females with bipolar disorder and epilepsy may cause menstrual cycle abnormalities, polycystic ovary syndrome, and hyperandrogenism. The effects on male reproductive functions have been researched only in epileptic patients and animals, and the results have been conflicting, because reproductive function abnormalities may be independent of the use of valproate and may be due to epilepsy itself. In the first part of this review reproductive function abnormalities due to epilepsy will be discussed, independent of the use of valproate or any other antiepileptic. Then, the results of research on valproate's effects on male reproductive functions (hormonal levels and sperm parameters) will be presented, including the possible underlying mechanisms of these effects, the effect of the duration of valproate use, and whether or not the effects are reversible. In the second section we review the results of animal research, which could be beneficial in assessing the effects of valproate and epilepsy. PMID- 20013430 TI - [BDNF measurement in stress-related mood disorders: a review of clinical studies]. AB - OBJECTIVE: The study of brain-derived neurotrophic factor (BDNF) in mood disorders and other psychiatric disorders is increasing. Of particular interest is whether or not BDNF can be a marker for psychiatric disorders. The aim of this study was to review the published findings on BDNF in stress-related mood disorders. METHOD: Searches were conducted of the PubMed and Psikiyatri Dizini databases to access international and national publications, respectively. Database searches were conducted with the keywords BDNF, and stress and mood disorders. RESULTS: BDNF plays a role in the physiopathology of stress-related changes and is supposedly related to a decrease in the number and size of neurons. Additionally, it has been suggested that serum levels of BDNF are lower in depressive patients than in the healthy controls. Nonetheless, BDNF levels increase after antidepressant treatment, along with symptom recovery, to those seen in healthy control subjects. This increase cannot be achieved in patients that do not respond to antidepressant treatment. On the other hand, in patients with stress-related non-depressive psychiatric disorders, serum BDNF is also low. CONCLUSION: To date, even though consistent findings of reduced BDNF levels have been reported, the specificity of these findings is doubtful. Serum BDNF might be considered a marker for stress-related mood disorders. PMID- 20013431 TI - [Two stiff person cases misdiagnosed as conversion disorder]. AB - Modern psychiatric diagnostic systems classify neurological symptoms that cannot be explained by a physical disease or another psychiatric disorder as conversion disorder (CD) or dissociative motor disorder. It is a well-known fact that the overall rate of misdiagnosis of conversion symptoms is high. The most common presenting symptoms of misdiagnosed patients are gait and movement disturbances. Stiff-person syndrome (SPS) is a rare progressive autoimmune neurological disorder. The identification of antibodies against glutamic acid decarboxylase (GAD) in association with SPS provided an important contribution to the understanding of the pathophysiology of this syndrome. Patients may present with severe muscle rigidity and sudden contractions. Simultaneous contraction of agonist and antagonist muscles produces gait disturbance. SPS can be exacerbated by emotional stressors, and sudden auditory, visual, and tactile stimuli. Herein we present 2 patients that were referred for psychiatric assessment, because their neurological symptoms initially could not be explained by a neurological disease, and subsequently diagnosed as SPS. The aim of this case report is to draw attention to the psychiatric presentations of SPS and to emphasize the importance of complete psychiatric and neurological examination, including brain imaging and electrophysiological studies, in the differential diagnosis of CD. PMID- 20013432 TI - [Serotonin syndrome associated with linezolid use: a case report]. AB - The serotonin syndrome is a toxic state largely attributable to changes in sensitivity of serotonin receptor system in the brainstem and spinal cord resulting from increased serotonergic activity in central neurologic system, due to use of serotonergic agents either in overdose or in combination. Serotonin syndrome may present with neuromuscular (clonus, myoclonus, tremor, hyperreflexia) and autonomic (fever, mydriasis, tachycardia, tachypnea) symptoms and mental status changes (confusion, agitation) and may result in death in severe cases. The risk for the development of serotonin syndrome is increased with the combined use of agents from different groups such as selective serotonin reuptake inhibitors (SSRIs) and monoamine oxidase inhibitors (MAOIs). The growing use of SSRIs for depression and the introduction of pharmacological agents newly developed for the treatment of various medical disorders increases the risk of drug-drug interactions and toxic states like serotonin syndrome. In the presented case clinical presentation and outcome of the serotonin syndrome which has developed as a consequence of concomitant linezolid use in a young patient who was already on an SSRI antidepressant is discussed. Linezolid is an oxazolidinone antibiotic which has MAOI-like properties. This case is presented to inform psychiatrists especially working in consultation-liaison settings about the risk of drug-drug interactions and possible prevention of these. PMID- 20013433 TI - [Acute prostatism associated with milnacipran therapy: a case report]. AB - Milnacipran is a dual serotonin (5-HT) and noradrenaline (NA) reuptake inhibitor. Milnacipran is an orally administered drug and indicated for the treatment of major depressive disorder. It is a safe and well tolerated drug for use in daily clinical practice. Prostatism; a symptom complex resulting from compression or obstruction of the urethra symptoms include diminution in the calibre and force of the urinary stream, hesitancy in initiating voiding, inability to terminate micturition abruptly, sensation of incomplete bladder emptying and rarely urinary retention. A 38 year old male patient with major depression was prescribed milnacipran 25 mg bid. Within 24 hours after with the second dose the patient developed hesitancy in initiating voiding, intermittency, terminal dribbling, projection disability, calibration disability and incomplete emptying. Due to these side effects patient has discontinued the drug. These side effects regress and disappeared within 24 hours. He had no past and familial history of any illness. Since there was neither a concomitant medication nor a medical illness, in addition to the absence of physical, laboratory and radiological evidence of a possible cause for acute prostatism an association between milnacipran was suggested. In this report, we identify clinical characteristics of milnacipran induced acute prostatism. Physicians should be aware of this possible adverse effect. Patients should be informed about these signs and symptoms. Although milnacipran may cause on urinary complaints, this side effect may be useful for patients urine incontinence. PMID- 20013434 TI - Simulating posterior parietal damage in a biologically plausible framework: neuropsychological tests of the search over time and space model. AB - The search over time and space (sSoTS) model attempts to simulate both the spatial and the temporal aspects of human visual search using spiking level neurons, which incorporate some biologically plausible aspects of neuronal firing. The model contains pools of units that (a) code basic features of objects, presumed to reside in the ventral visual stream, and (b) respond in a feature-independent way to stimulation at their location, presumed to operate in the posterior parietal cortex. We examined the effects of selective lesioning neurons responding to one side of the location map. Unilateral damage introduced spatial biases into selection that affected conjunction more than single-feature search. In addition, there was an impaired ability to segment stimuli over time as well as space (e.g., in preview search). These results match previously reported data on patients with posterior parietal lesions. In addition we show that spatial biases in selection increase under conditions in which there is decreased activity from excitatory neurotransmitters, mimicking effects of reduced arousal. Further simulations explored the effects of time and of visual grouping on extinction, generating predictions that were then tested empirically. The model provides a framework for linking behavioural data from patients with neural-level determinants of visual attention. PMID- 20013436 TI - Assessment of head injury of children due to golf ball impact. AB - Head trauma injury due to impact by a flying golf ball is one of the most severe possible injury accidents on the golf course. Numerical simulations based on the finite element method are presented to investigate head injury in children due to impact by a flying golf ball. The stress and energy flow patterns in a head model during the golf ball impact are computed for various combinations of striking speed, falling angle of the golf ball before impact, and impact location. It is found that a child is more prone to head injury due to golf ball impact on the frontal and side/temporal areas. The simulated results are found to conform to the clinical reports on children's head injuries from flying golf balls. PMID- 20013435 TI - Finite element modelling of the glenohumeral capsule can help assess the tested region during a clinical exam. AB - The objective of this research was to examine the efficacy of evaluating the region of the glenohumeral capsule being tested by clinical exams for shoulder instability using finite element (FE) models of the glenohumeral joint. Specifically, the regions of high capsule strain produced by glenohumeral joint positions commonly used during a clinical exam were identified. Kinematics that simulated a simple translation test with an anterior load at three external rotation angles were applied to a validated, subject-specific FE model of the glenohumeral joint at 60 degrees of abduction. Maximum principal strains on the glenoid side of the inferior glenohumeral ligament (IGHL) were significantly higher than the maximum principal strains on the humeral side, for all three regions of the IGHL at 30 degrees and 60 degrees of external rotation. These regions of localised strain indicate that these joint positions might be used to test the glenoid side of the IGHL during this clinical exam, but are not useful for assessing the humeral side of the IGHL. The use of FE models will facilitate the search for additional joint positions that isolate high strains to other IGHL regions, including the humeral side of the IGHL. PMID- 20013437 TI - Optimal applicator placement in hepatic radiofrequency ablation on the basis of rare data. AB - In this paper, a numerical procedure to determine an optimal applicator placement for hepatic radiofrequency ablation incorporating uncertain material parameters is presented. The main focus is set on the treatment of subjective and rare data based information. For this purpose, we employ the theory of fuzzy sets and model uncertain parameters as fuzzy quantities. While fuzzy modelling has been established in structural engineering in the recent past, it is novel in biomedical engineering. Incorporating fuzzy quantities within an optimisation task is basically innovative. In our context, fuzzy modelling allows us to determine an optimal applicator placement that maximises the therapy success under the given uncertainty conditions. The applicability of our method is demonstrated by means of an example case. PMID- 20013438 TI - Perceptions of Racism in Children and Youth (PRaCY): properties of a self-report instrument for research on children's health and development. AB - Experiences of racial discrimination have been demonstrated to be related to racial and ethnic disparities in mental and physical health and healthcare. There has been little study, however, of the relationship between racism and health in children, and few psychometrically valid and reliable instruments to measure Perceptions of Racism in Children and Youth (PRaCY) exist. This paper reports on the development and testing of such an instrument, the PRaCY. Development of the instrument began with open-ended qualitative interviews, from which a proto questionnaire was created. The questionnaire gathered information on the prevalence, attribution, emotional responses, and coping responses to 23 situations identified by participants in the qualitative phase. The proto questionnaire was administered to 277 children between the ages of 8 and 18 years (38% Latino/a, 31% African-American, 19% multiracial/multicultural, 7% West Indian/Caribbean, and 5% Other). Item analysis resulted in two developmentally appropriate 10-item instruments (one for ages 7-13, another for ages 14-18). Internal consistency reliability was strong (alpha = 0.78 for both versions of the instrument). Confirmatory factor analysis demonstrated good fit for both versions (younger-Comparative Fit Index (CFI): 0.967, Root Mean Square Error of Approximation (RMSEA): 0.047; older-CFI: 0.934, RMSEA: 0.056). Differential item functioning analyses demonstrated no group-specific biases in item response. PRaCY scores were appropriately associated with higher depressive symptom scores and elevated anxiety scores in the younger sample. Results indicate that the PRaCY is a valid and reliable instrument that measures perceptions of racism and discrimination in children and youth aged 8-18 from diverse racial/ethnic backgrounds. PMID- 20013439 TI - Is pregnancy a teachable moment for smoking cessation among US Latino expectant fathers? A pilot study. AB - OBJECTIVE: Pregnancy may be a time when US Latino expectant fathers consider quitting smoking. A 'teachable moment' is theorized to increase motivation to change a behavior through increased risk perceptions, emotional responses, and changes in self-image. DESIGN: We recruited 30 Spanish-speaking expectant fathers through their pregnant partners. We assessed expectant fathers' diet, exercise, and smoking and teachable moment constructs (risk perceptions, emotional responses, and self-image).We also tested correlations between teachable moment constructs and motivation to change behaviors. RESULTS: Latino expectant fathers had high-risk perceptions that their smoking harmed the pregnancy (M=4.4, SD=0.5 on five-point scale) and strong emotional responses about their smoking during pregnancy (M=3.9, SD=1.1). They also felt it was their role to make the pregnancy healthy (M=4.4, SD=0.8). They felt less strongly that their diet and exercise affected the pregnancy. The teachable moment constructs for smoking were strongly correlated with motivation to quit smoking; the same was not true for diet and exercise. CONCLUSIONS: Latino expectant fathers seem aware that their smoking could harm the pregnancy but seem less concerned about the effect of their diet and exercise on the pregnancy. Pregnancy may be a time to help Latino expectant fathers quit smoking. PMID- 20013440 TI - The development and initial validation of the Hawaiian Youth Drug Offers Survey (HYDOS). AB - OBJECTIVE: This study describes the development and preliminary validation of a survey focused on the most salient situations where drugs and/or alcohol are offered to Native Hawaiian youth in rural communities. DESIGN: The study used a five-phase approach to test development and validation. In Phase 1 (item generation), survey items were created from a series of focus groups with middle school aged youth (n = 47). In Phase 2 (item refinement and selection), items were edited and reduced to 62 drug-offer situations that were selected for inclusion in the survey. In Phase 3 (item reduction), items were administered to 249 youth from seven middle or intermediate schools in Hawai'i. RESULTS: Exploratory factor analysis of the Native Hawaiian subsample (n = 194) indicated the presence of three factors accounting for 63% of the variance: peer pressure (23%); family offers and context (21%); and unanticipated drug offers (19%). The survey items differentiated between Hawaiian and non-Hawaiian youth respondents, supporting the validity of the questionnaire. The hypothesized relationship between cultural connectedness and drug offer exposure was not confirmed. Internal consistency of the measure was high. CONCLUSIONS: The survey helps to fill the gap in information related to drug use of Native Hawaiian youth and has implications for the development and assessment of culturally-specific drug prevention programs for these youth. PMID- 20013442 TI - Analysis of food taints and off-flavours: a review. AB - Taints and off-flavours in foods are a major concern to the food industry. Identification of the compound(s) causing a taint or off-flavour in food and accurate quantification are critical in assessing the potential safety risks of a product or ingredient. Even when the tainting compound(s) are not at a level that would cause a safety concern, taints and off-flavours can have a significant impact on the quality and consumers' acceptability of products. The analysis of taints and off-flavour compounds presents an analytical challenge especially in an industrial laboratory environment because of the low levels, often complex matrices and potential for contamination from external laboratory sources. This review gives an outline of the origins of chemical taints and off-flavours and looks at the methods used for analysis and the merits and drawbacks of each technique. Extraction methods and instrumentation are covered along with possible future developments. Generic screening methods currently lack the sensitivity required to detect the low levels required for some tainting compounds and a more targeted approach is often required. This review highlights the need for a rapid but sensitive universal method of extraction for the unequivocal determination of tainting compounds in food. PMID- 20013441 TI - Pattern of intake of food additives associated with hyperactivity in Irish children and teenagers. AB - A double-blind randomized intervention study has previously shown that a significant relationship exists between the consumption of various mixes of seven target additives by children and the onset of hyperactive behaviour. The present study set out to ascertain the pattern of intake of two mixes (A and B) of these seven target additives in Irish children and teenagers using the Irish national food consumption databases for children (n = 594) and teenagers (n = 441) and the National Food Ingredient Database. The majority of additive-containing foods consumed by both the children and teenagers contained one of the target additives. No food consumed by either the children or teenagers contained all seven of the target food additives. For each additive intake, estimates for every individual were made assuming that the additive was present at the maximum legal permitted level in those foods identified as containing it. For both groups, mean intakes of the food additives among consumers only were far below the doses used in the previous study on hyperactivity. Intakes at the 97.5th percentile of all food colours fell below the doses used in Mix B, while intakes for four of the six food colours were also below the doses used in Mix A. However, in the case of the preservative sodium benzoate, it exceeded the previously used dose in both children and teenagers. No child or teenager achieved the overall intakes used in the study linking food additives with hyperactivity. PMID- 20013443 TI - Concentrations of undeclared allergens in food products can reach levels that are relevant for public health. AB - Food products can become contaminated with food allergens due to cross-contact. Precautionary 'may contain' labelling may alert to the possible presence of an allergen, but guidance for such labelling is lacking. As a result, allergy information on the packaging may not be reliable and allergic consumers might be at risk of allergic reactions after consuming unlabelled, but indeed contaminated, products. Recently, a cow's milk protein allergic patient experienced a severe allergic reaction to a dark chocolate product containing undeclared milk proteins. This case induced the authors to investigate to what extent allergen concentrations of unlabelled products reach levels that are of public health relevance. The concentrations of milk proteins in the complaint sample and a collection of products of other batches and brands purchased from different stores were analysed. Together with appropriate threshold and food consumption data, the risks of allergic reactions and the severity of these reactions within the adult milk-allergic population were determined using probabilistic risk assessment techniques. The results show that milk protein concentrations in unlabelled products reach levels that may elicit allergic reactions in up to 68% of the adult allergic consumers. Therefore, concentrations of allergens in unlabelled products could reach levels that are of public health relevance. Application of probabilistic risk assessment can be an aid in revealing the public health consequences of undeclared allergens in food, in risk management decision-making and developing guidance in terms of risk-based concentration limits for precautionary labelling. PMID- 20013444 TI - Determination of free and total sulfites in wine using an automatic flow injection analysis system with voltammetric detection. AB - An automated flow injection analysis (FIA) system, based on an initial analyte separation by gas-diffusion and subsequent determination by square-wave voltammetry (SWV) in a flow cell, was developed for the determination of total and free sulfur dioxide (SO(2)) in wine. The proposed method was compared with two iodometric methodologies (the Ripper method and a simplified method commonly used by the wine industry). The developed method displayed good repeatability (RSD lower than 6%) and linearity (between 10 and 250 mg l(-1)) as well as a suitable LOD (3 mg l(-1)) and LOQ (9 mg l(-1)). A major advantage of this system is that SO(2) is directly detected by flow SWV. PMID- 20013445 TI - Comparison of extraction solvents and conditions for herbicide residues in milled rice with liquid chromatography-diode array detection analysis (LC-DAD). AB - Different extraction procedures and clean-up methods were compared in order to develop a sample preparation procedure for the multi-residue analysis of six post emergence herbicides (metsulfuron methyl, bensulfuron methyl, pyrazosulfuron ethyl, bentazone, bispyribac sodium and cyhalofop butyl) in rice grains followed by liquid chromatography-diode array detection (LC-DAD). Optimum results were obtained dispersing milled rice grain in water, followed by the addition of 1% acetic acid in acetonitrile, MgSO(4) and sodium acetate as a modification of the quick, easy, cheap, effective, rugged and safe (QuEChERS) method but no primary and secondary amine (PSA) sorbent was added due to the acidic nature of the herbicides. The method was further expanded to other post-emergence herbicides (quinclorac, clomazone and propanil). Except for quinclorac, which cannot be analysed with this method, the recoveries of the other eight herbicides were in the range 73-111%, with relative standard deviations lower than 12%. Limits of detection (LODs) ranged from 0.03 to 0.08 mg kg(-1). A single analyst can extract twelve samples in 4 h. The method presented here allows the simultaneous residue determination of the most common post-emergence herbicides employed in cultivating rice. It is simple, rapid, sensitive, and can be applied routinely to polished rice grain herbicide residue analysis. PMID- 20013446 TI - Health risk assessment of ochratoxin A for all age-sex strata in a market economy. AB - In order to manage risk of ochratoxin A (OTA) in foods, we re-evaluated the tolerable daily intake (TDI), derived the negligible cancer risk intake (NCRI), and conducted a probabilistic risk assessment. A new approach was developed to derive 'usual' probabilistic exposure in the presence of highly variable occurrence data, such as encountered with low levels of OTA. Canadian occurrence data were used for various raw food commodities or finished foods and were combined with US Department of Agriculture (USDA) food consumption data, which included data on infants and young children. Both variability and uncertainty in input data were considered in the resulting exposure estimates for various age/sex strata. Most people were exposed to OTA on a daily basis. Mean adjusted exposures for all age-sex groups were generally below the NCRI of 4 ng OTA kg bw( 1), except for 1-4-year-olds as a result of their lower body weight. For children, the major contributors of OTA were wheat-based foods followed by oats, rice, and raisins. Beer, coffee, and wine also contributed to total OTA exposure in older individuals. Predicted exposure to OTA decreased when European Commission maximum limits were applied to the occurrence data. The impact on risk for regular eaters of specific commodities was also examined. PMID- 20013447 TI - Validation data for HPLC/FLD determinations of ochratoxin A in red paprika and black pepper adopting a one-step clean-up procedure. AB - Validation data for the determination of ochratoxin A (OTA) in two spice matrices, red paprika and black pepper, were obtained for samples prepared with a simplified single-step clean-up column. Extracts of finely ground samples of red paprika and black pepper were prepared and applied to a Mycosep 229 Ochra clean up column. The purified extract was then subjected to HPLC/FLD analysis. The relative standard deviation for repeatability (RSDr) of the method was 11.8% for red paprika and 9.9% for black pepper. The limit of detection (LOD) value (three times the noise) was estimated as corresponding to the response of an extract derived from a blank matrix (previously washed) and spiked at 1.0 microg kg(-1). The limit of quantitation (LOQ) (three times LOD) was 3.0 microg kg(-1). The performance of the one-step column clean-up procedure appears to be a suitable alternative to commonly used clean-up techniques and allows the precise determination of OTA in two complex matrices. PMID- 20013448 TI - Urine metabolite analysis as a function of deoxynivalenol exposure: an NMR-based metabolomics investigation. AB - Deoxynivalenol (DON) is a toxic fungal metabolite that frequently contaminates cereal crops including wheat, maize and barley. Despite knowledge of frequent exposure through diet, our understanding of the potential consequences of human exposure remains limited, in part due to the lack of validated exposure biomarkers. In this study, we interrogated the urinary metabolome using nuclear magnetic resonance (NMR) spectroscopy to compare individuals with known low and high DON exposure through consumption of their normal diet. Urine samples from 22 adults from the UK (seven males, 15 females; age range = 21-59 years) had previously determined urinary DON levels using an established liquid chromatography-mass spectrometry (LC-MS) assay. Urine samples were subsequently analysed using an NMR-based metabolomics approach coupled with multivariate statistical analysis. Metabolic profiling suggested that hippurate levels could be used to distinguish between groups with low (3.6 ng DON mg(-1) creatinine: 95% CI = 2.6, 5.0 ng mg(-1)) and high (11.1 ng mg(-1): 95% CI = 8.1, 15.5 ng mg(-1)) DON exposure, with the concentration of hippurate being significantly (1.5 times) higher for people with high DON exposure than for those with low DON exposure (p = 0.047). This, to our knowledge, is the first report of a metabolomics-derived biomarker of DON exposure in humans. PMID- 20013449 TI - Test procedures for obtaining representative extracts suitable for reliable in vitro toxicity assessment of paper and board intended for food contact. AB - This paper describes the use of a suite of extraction procedures applicable to the assessment of the in vitro toxicity of paper/board samples intended for food contact applications. The sample is extracted with ethanol, water, or exposed to modified polyphenylene oxide (Tenax) for fatty, non-fatty and dry food applications, respectively. The water extracts are directly suitable for safety assessment using in vitro bioassays. The ethanol extracts of the paper/board and of the exposed Tenax require pre-concentration to give acceptable sensitivity. This is because the in vitro bioassays can tolerate only a small percentage of added organic solvent before the solvent itself inhibits. The extraction procedures have been selected such that they mimic the foreseeable conditions of use with foods and that they are also fully compatible with the battery of in vitro biological assays for the safety assessment of the total migrate. The application of the extraction protocols is illustrated by the results for one of the many paper/board samples provided by the BIOSAFEPAPER project industrial platform members. The assessment indicated that this sample should not be considered as suitable for use with fatty foodstuffs but was suitable for dry and non-fatty foods. Information subsequently received from the manufacturer revealed that this was a non-food-grade product included in the project to test the capabilities of the bioassay procedures. The selection criteria for the test conditions and the suite of methods developed have been prepared in Comite Europeen de Normalisation (CEN) format and is currently being progressed by CEN/TC172 as a European Standard. PMID- 20013450 TI - Manganese, iron, and total particulate exposures to welders. AB - Welders are exposed to a variety of metal fumes, including manganese, that may elevate the risk for neurological disease. This study examines several large data sets to characterize manganese, iron, and total particulate mass exposures resulting from welding operations. The data sets contained covariates for a variety of exposure modifiers, including the presence of ventilation, the degree of confinement, and the location of the personal sampler (i.e., behind or in front of the welding helmet). The analysis suggests that exposures to manganese are frequently at or above the current ACGIH(R) threshold limit value of 0.2 mg/m(3). In addition, there is evidence that local exhaust ventilation can control the exposures to manganese and total fume but that mechanical ventilation may not. The data suggest that higher exposures are associated with a greater degree of enclosure, particularly when local exhaust ventilation is absent. Samples taken behind the helmet were, in general, lower than those measured outside of it. There were strong correlations among manganese, iron, and total particulate mass exposures, suggesting simple equations to estimate one fume component from any of the others. PMID- 20013451 TI - The double-entry intraclass correlation as an index of profile similarity: meaning, limitations, and alternatives. AB - As applied to many psychological phenomena, analysis of profile similarity has an intuitive appeal that masks complex statistical issues. Psychologists have long debated the methods of indexing similarity between 2 psychological profiles, but the double-entry intraclass correlation may be emerging as a preferred approach. Unfortunately, the double-entry intraclass correlation has not been articulated clearly in terms of fundamental facets of profiles--elevation, scatter, and shape -which prevents full understanding of its meaning. In this article, I (a) articulate these effects, (b) discuss potential limitations and confusions arising from these effects, (c) present a failure to replicate previous empirical findings regarding the double-entry intraclass correlation, and (d) present alternative recommendations for analysis of profile similarity. The conceptual, mathematical, and empirical points may enhance the insights emerging from analyses of profiles and profile similarity. PMID- 20013452 TI - The Multidimensional Perfectionism Cognitions Inventory-English (MPCI-E): reliability, validity, and relationships with positive and negative affect. AB - The Multidimensional Perfectionism Cognitions Inventory (MPCI; Kobori & Tanno, 2004) is a promising new instrument developed in Japan to assess perfectionism cognitions regarding personal standards, pursuit of perfection, and concern over mistakes. In this study, we examined reliability and validity of the English version of the MPCI, the MPCI-E (Kobori, 2006), in a sample of 371 native English speakers. A confirmatory factor analysis confirmed the MPCI-E's 3-factorial oblique structure. Moreover, correlations with measures of dispositional perfectionism and past-week positive and negative affect provided initial evidence of the MPCI-E's convergent and differential validity. Finally, hierarchical multiple regressions indicated that the MPCI-E showed incremental validity in explaining variance in positive and negative affect above variance explained by dispositional perfectionism. Overall, the findings provide initial evidence for the reliability and validity of the MPCI-E as a multidimensional measure of perfectionism cognitions that has the potential to further the understanding of positive and negative cognitions in perfectionism. PMID- 20013453 TI - The Chinese Parental Stress Scale: psychometric evidence using Rasch modeling on clinical and nonclinical samples. AB - In this study, we examined the psychometric properties of the Parental Stress Scale (PSS; Berry & Jones, 1995) with Chinese parents. Participants included 162 parents recruited through primary schools and 38 parents with children with attention deficit hyperactivity disorder (ADHD). Rasch analysis results indicated that 2 items were outside the accepted fit statistics range and that the Likert scale response categories were not functioning properly. We identified a reduced 16-item scale with 5 categories as unidimensional, with a person reliability of .86; the Likert scale response categories worked well. The revised PSS correlated with measures of child behavior and parenting stress and could differentiate between the 2 groups of parents. PMID- 20013454 TI - Structured Interview of Personality Organization (STIPO): preliminary psychometrics in a clinical sample. AB - In this article, we describe the development and preliminary psychometric properties of the Structured Interview of Personality Organization (STIPO), a semistructured interview designed for the dimensional assessment of identity, primitive defenses, and reality testing, the three primary content domains in the model of personality health and disorder elaborated by Kernberg (1984; Kernberg & Caligor, 2005). Results of this investigation, conducted in a clinical sample representing a broad range of personality pathology, indicate that identity and primitive defenses as operationalized in the STIPO are internally consistent and that interrater reliability for all 3 content domains is adequate. Validity findings suggest that the assessment of one's sense of self and significant others (Identity) is predictive of measures of positive and negative affect, whereas the maladaptive ways in which the subject uses his or her objects for purposes of regulating one's self experience (Primitive Defenses) is predictive of measures of aggression and personality disorder traits associated with cluster B personality disorders. We discuss implications of these findings in terms of the theory-driven and trait-based assessment of personality pathology. PMID- 20013455 TI - The Anxiety Sensitivity Index-3: analyses of dimensions, reliability estimates, and correlates in nonclinical samples. AB - We investigated the factor structure, reliability estimates, and correlates of the Anxiety Sensitivity Index-3 (ASI-3; Taylor et al., 2007) in 2 studies. We established a bifactor model in Study 1 as an alternative representation of the structure of the ASI-3. Analyses of gender differences on the total ASI-3 and subscale scores were not statistically significant (Study 1, N = 462). In Study 2 (N = 293), results of a series of confirmatory factor analyses provided stronger support for the fit of the bifactor model compared with 2 alternative models. Estimates of scale reliability were adequate (all rho values > or = .80) and not "p" (as in italic p for significance). in the 2 studies. In addition, using simultaneous regression analyses, we found anxiety-specific correlates for the total ASI-3 and subscale scores to include responses on self-report measures of interpersonal sensitivity, obsessive-compulsive anxiety, paranoid ideation, and phobic anxiety. PMID- 20013456 TI - Construct validity of the Social Physique Anxiety Scale in a French adolescent sample. AB - We conducted a series of studies to investigate the psychometric properties of the Social Physique Anxiety Scale (SPAS; Hart, Leary, & Rejeski, 1989) among a total sample of 1,563 nonclinical French adolescents. The, first study provided support for the item content of the preliminary version of the French SPAS for adolescents. Then, the second study supported the convergent validity of the English and French version of the SPAS. Finally, Studies 3 to 6 provided support for the factor validity, measurement invariance (across genders, age groups and samples), latent mean structure invariance (across age category and samples), reliability, convergent validity, and criterion-related validity of a 7-item version of the SPAS. PMID- 20013457 TI - Longitudinal validation of the urgency traits over the first year of college. AB - Research has identified 2 emotion-based dispositions to rash action, referred to as positive urgency and negative urgency. They are thought to reflect tendencies to engage in rash acts when in extremely positive and extremely negative moods, respectively. In this article, we describe the first direct test of this hypothesis. We measured the urgency traits and risky behavior involvement while in extremely positive and negative moods over the course of the first year of college for 292 students. After controlling for sex, typical mood state, and prior mood-based rash behavior, positive urgency predicted increases in positive mood-based rash action, and negative urgency predicted increases in negative mood based rash action during the first year of college. These results provide further evidence for the validity of the theory of emotion-based rash action and for the measures of positive and negative urgency. PMID- 20013458 TI - Psychometric properties of a Spanish version of the UPPS-P impulsive behavior scale: reliability, validity and association with trait and cognitive impulsivity. AB - Impulsivity is a multifaceted construct central to several forms of psychopathology. Recently, Lynam, Smith, Whiteside, and Cyders (2006) developed the UPPS-P scale, a multidimensional inventory that assesses 5 personality pathways contributing to impulsive behavior: negative urgency, lack of perseverance, lack of premeditation, sensation seeking, and positive urgency. In this study, we aimed (a) to analyze the psychometric properties of a Spanish version of the UPPS-P scale and (b) to explore the relationship between the different dimensions of the UPPS-P scale and conceptually related constructs including trait measures derived from different models of impulsive personality (the Gray's [1987] and Plutchik's [1984] models) and a state measure of cognitive impulsivity, the Delay-Discounting Test (Kirby, Petry, & Bickel, 1999). We administered the UPPS-P scale along with the Sensitivity to Punishment and Sensitivity to Reward Questionnaire (Torrubia, Avila, Molto, & Caseras, 2001), the Plutchik Impulsivity Scale (Plutchik & Van Praag, 1989), and the Delay Discounting Test to a sample of 150 undergraduate students. Results showed that the Spanish adaptation of the UPPS-P scale have appropriate psychometric properties. Different dimensions of the UPPS-P were differentially associated with predicted conceptually related constructs. We conclude that the Spanish adaptation of the UPPS-P scale is a useful instrument for fine-grained assessment of impulsivity in Spanish-speaking adult population. PMID- 20013459 TI - What might this be? Rediscovering the Rorschach as a tool for personnel selection in organizations. AB - In this article, I review the empirical status of the Rorschach as it relates to potential use of the instrument in personnel selection procedures. As part of this review, I outline developing trends in personnel selection practices, discuss the Rorschach literature relevant to organizational performance, and weigh the benefits and drawbacks of using the instrument in this capacity. Based on this analysis, I argue that the Rorschach may represent a unique and potentially valuable tool for assessing personality as part of comprehensive personnel selection procedures. I conclude with a proposed trajectory for Rorschach research intended to better determine the instrument's viability in organizational settings. PMID- 20013460 TI - Relationships between maturity status, physical activity, and physical self perceptions in primary school children. AB - The aim of this study was to examine the influence of maturity status on primary school children's physical activity and physical self-perceptions. Altogether, 175 children (97 girls, 78 boys) aged 10.6 +/- 0.3 years completed the Children and Youth Physical Self-Perception Profile and wore an ActiGraph accelerometer for five consecutive days to assess moderate-to-vigorous physical activity. Anthropometric measures were completed to estimate maturity status. A two-level, multi-level analysis was used to assess the influence of maturity status on moderate-to-vigorous physical activity and physical self-perceptions. Boys performed more moderate-to-vigorous physical activity than girls (P < 0.0001), but when the effect of maturity status was controlled the difference was reduced (P = 0.02). Significant differences between the sexes were also observed for physical self-perception sub-domains (boys > girls, P = 0.02 to 0.0001). When maturity status was added to the model, significant differences were no longer apparent for each sub-domain, with the exception of perceived strength. Significant interactions between gender and maturity status revealed that boys' physical self-perceptions improved with more advanced maturity status, whereas girls' self-perceptions decreased (P = 0.07 to 0.002). Significant differences between the sexes in moderate-to-vigorous physical activity and some domains of physical self-perceptions were reduced or no longer evident when the effect of maturity status was controlled. Maturity status may differentially influence boys' and girls' physical self-perceptions. PMID- 20013461 TI - Movement patterns in cricket vary by both position and game format. AB - We compared the movement patterns of cricketers in different playing positions across three formats of cricket (Twenty20, One Day, multi-day matches). Cricket Australia Centre of Excellence cricketers (n = 42) from five positions (batting, fast bowling, spin bowling, wicketkeeping, and fielding) had their movement patterns (walk, jog, run, stride, and sprint) quantified by global positioning system (GPS) technology over two seasons. Marked differences in movement patterns were evident between positions and game formats, with fast bowlers undertaking the greatest workload of any position in cricket. Fast bowlers sprinted twice as often, covered over three times the distance sprinting, with much smaller work-to recovery ratios than other positions. Fast bowlers during multi-day matches covered 22.6 +/- 4.0 km (mean +/- s) total distance in a day (1.4 +/- 0.9 km in sprinting). In comparison, wicketkeepers rarely sprinted, despite still covering a daily total distance of 16.6 +/- 2.1 km. Overall, One Day and Twenty20 cricket required approximately 50 to 100% more sprinting per hour than multi-day matches. However, multi-day cricket's longer duration resulted in 16-130% more sprinting per day. In summary, the shorter formats (Twenty20 and One Day) are more intensive per unit of time, but multi-day cricket has a greater overall physical load. PMID- 20013462 TI - Adolescent girls' energy expenditure during dance simulation active computer gaming. AB - The objective of this study was to determine the energy expended and intensity of physical activity achieved by adolescent girls while playing on a dance simulation game. Twenty adolescent girls were recruited from a local secondary school. Resting oxygen uptake (VO(2)) and heart rate were analysed while sitting quietly and subsequently during approximately 30 min of game play, with 10 min at each of three increasing levels of difficulty. Energy expenditure was predicted from VO(2) at rest and during game play at three levels of play, from which the metabolic equivalents (METS) of game playing were derived. Mean +/- standard deviation energy expenditure for levels 1, 2, and 3 was 3.63 +/- 0.58, 3.65 +/- 0.54, and 4.14 +/- 0.71 kcal . min(-1) respectively, while mean activity for each level of play was at least of moderate intensity (>3 METS). Dance simulation active computer games provide an opportunity for most adolescent girls to exercise at moderate intensity. Therefore, regular playing might contribute to daily physical activity recommendations for good health in this at-risk population. PMID- 20013463 TI - Eccentric exercise affects the upper limbs more than the lower limbs in position sense and reaction angle. AB - In this study, we investigated the effect of eccentric exercise on position sense and reaction angle of the elbow and knee flexors. Twelve males underwent two eccentric exercise sessions involving a randomized crossover design. In the first session participants used their elbow flexors and in the other session their knee flexors. Muscle damage indices, position sense, and joint reaction angle to release of the elbow and knee flexors were measured before, immediately after, and up to 7 days after exercise. Exercise induced greater muscle damage in the elbow flexors than knee flexors. Exercise disturbed position sense of the elbow and knee joint. For both limbs, the participants adopted a more extended position than the reference angle. The elbow and knee joint reaction angles to release increased after exercise for both the elbow and knee flexors. The disturbances in position sense and reaction angle after exercise were greater in the elbow flexors than knee flexors. The elbow flexors remained more accurate and faster than the knee flexors at all time points. These results may be explained by the higher density of muscle spindles and the lower innervation ratio of the elbow flexors compared with the knee flexors, as well as the fact that the arms are more accustomed than the legs to perform fast and accurate movements. PMID- 20013464 TI - The effects of age on remembering and knowing misinformation. AB - Previous research has suggested that older adults are more susceptible to misleading information. The current experiments examined the nature of older and younger participants' conscious experience of contradictory and additive misinformation (Experiment 1), and misinformation about a memorable or non memorable item (Experiment 2). Participants watched a video of a burglary before answering questions about the event that contained misinformation. Participants then completed a cued recall task whereby they answered questions and indicated whether they remembered the item, knew the item, or were guessing. The results indicated that older adults were less likely to remember or know the original item in comparison to younger adults but were also more likely to know misinformation than younger adults. This pattern occurred for contradictory misinformation and misleading information about memorable and non-memorable items. Only additive misinformation was associated with more remember responses for older but not younger adults. PMID- 20013465 TI - Oxidation of lynestrenol by the fungus Cunninghamella elegans. AB - Transformation of lynestrenol (19-nor-17alpha-pregn-4-en-20-yn-17beta-ol) (1) was carried out by incubation with Cunninghamella elegans to obtain 19-nor-17alpha pregn-4-en-20-yn-3-one-10beta,17beta-diol (2), 19-nor-17alpha-pregn-4-en-20-yn-3 one-6beta,17beta-diol (3), and 19-nor-17alpha-pregn-4-en-20-yn-3beta,6beta,17beta triol (4). Metabolite 4 was identified as a new compound. These metabolites were structurally characterised on the basis of spectroscopic techniques. PMID- 20013466 TI - Further anti-feedant neo-clerodanes from Teucrium tomentosum. AB - Reinvestigation of the acetone extract of Teucrium tomentosum yielded a new neo clerodane diterpenoid, teuctomin (1), along with 6-O-acetyl-teucmicropodin (2), teugracilin-A (3), 6alpha-hydroxyteuscordin (4) and 6alpha-acetoxyteuscordin (5). The structure of the new compound was elucidated comprehensively using 1D and 2D NMR methods. Compounds showed effective antifeedancy against Plutella xylostella at 10 microg cm(-2) of leaf area. PMID- 20013467 TI - A new dinorclerone diterpenoid glycoside from Tinospora sinensis. AB - A new dinorclerone diterpenoid glycoside, named 1-deacetyltinosposide A (1), was isolated from the stem of Tinospora sinensis together with 10 known compounds. Their structures were elucidated on the basis of extensive spectroscopic techniques (MS, IR, 1D and 2D NMR experiments). PMID- 20013469 TI - Monitoring the enzymatic degradation of sinigrin from B. juncea meal using (1)H NMR spectroscopy. AB - A simple procedure for extracting and purifying sinigrin from Oriental mustard (Brassica juncea) meal using cold water was developed. Subsequently, the chemical degradation of sinigrin due to enzymatic activity was monitored using (1)H NMR spectroscopy over the course of 30 h. The experimental findings show that sinigrin has been positively identified from B. juncea using (1)H NMR spectroscopy and the prescribed preparation procedure. The results that have been presented also offer unambiguous evidence that (1)H NMR spectroscopy can be used to detect changes in sinigrin concentration over time. PMID- 20013468 TI - Effect of successive increase in alcohol chains on reaction with isocyanates and isothiocyanates. AB - The reaction of isocyanates and isothiocyanates with long-chain alcohols, e.g. n hexanol, n-heptanol and n-octanol, exclusively gave N-aryl-O-alkyl carbamates, while N-aryl-O-alkyl carbamates were formed along with symmetrical 1,3 disubstituted ureas and thioureas when the same reactions were carried out with small-chain alcohols at room temperature without using any solvent. PMID- 20013470 TI - A new analogue of fatty alcohol from Tamarix hampeana L. AB - New analogues of a long-chain secondary alcohol (1) and laserine (2) were isolated from the flowers of Tamarix hampeana L. The isolated compounds were identified using 1D and 2D NMR, LCMS/APCI, and chemical methods. Laserine was isolated for the first time from T. hampeana L. PMID- 20013471 TI - Studies on chemical constituents of the leaves of Smallantus sonchifolius (yacon): structures of two new diterpenes. AB - The extract from the leaves of Smallantus sonchifolius (yacon) was found to show potent anti-diabetic activity. Two new diterpenes, named ent-kaurane 3beta,16beta,17, 19-tetrol (1) and ent-kaurane-16beta,17,18,19-tetrol (2), were isolated from the extract, together with six known compounds. The structures of the new compounds were determined on the basis of chemical and physicochemical evidence. PMID- 20013472 TI - Content and composition of the essential oil of Chamomilla recutita (L.) Rauschert from some European countries. AB - Variations in the essential oil composition of Chamomilla recutita (L.) Rauschert from different European countries were determined. A total of 39 components were identified, representing over 92% of the total oil yield. The principal biologically active compounds in chamomile oils were bisabolol oxide A (3.1 56.0%), alpha-bisabolol (0.1-44.2%), bisabolol oxide B (3.9-27.2%), cis-enyne bicycloether (8.8-26.1%), bisabolon oxide A (0.5-24.8%), chamazulene (0.7-15.3%), spathulenol (1.7-4.8%) and (E)-beta-farnesene (2.3-6.6%). In 8 chamomile samples from 13, bisabolol oxide A (27.5-56.0%) was predominant (among them in three Estonian samples). alpha-Bisabolol (23.9-44.2%) was predominant in the samples from Moldova, Russia and the Czech Republic. The sample from Armenia was rich in bisabolol oxide B (27.2%) and chamazulene (15.3%). The oils were obtained in yields of 0.7-6.7 mL kg(-1) and the minimum limit of 4 mL kg(-1) stated by the European Pharmacopoeia was exceeded only in 13 samples from 13 analysed drugs. PMID- 20013473 TI - Two new 3-benzylphthalides from Scorzonera veratrifolia Fenzl. AB - A phytochemical investigation of Scorzonera veratrifolia Fenzl (Asteraceae) yielded two new 3-benzylphthalides, scorzoveratrin (1) and scorzoveratrozit (2), together with five known compounds, chlorogenic acid (3), chlorogenic acid methyl ester (4), cryptochlorogenic acid (5), 4,5-dicaffeoylquinic acid (6) and 3,5 dicaffeoylquinic acid (7). All secondary metabolites were assigned using physicochemical and spectroscopic data. The known compounds 3-7 were isolated for the first time from this species. PMID- 20013475 TI - Sesqui- and diterpenoids from three New Zealand liverworts: Bazzania novae zelandiae, Gackstroemia sp. and Dendromastigophora sp. AB - Two new sesquiterpenoids, 1 and 2, were isolated from the unidentified liverwort Gackstroemia species, and three known sesquiterpenoids, 3-5, and a known sesqui- (6) and diterpenoid (7) were isolated from Bazzania novae-zelandiae and the unidentified Dendromastigophora species, respectively. Their structures were determined by spectroscopic analysis. PMID- 20013474 TI - A new benzylisoquinoline alkaloid from Argemone mexicana. AB - A new benzylisoquinoline alkaloid, argemexirine, together with two known protoberberine alkaloids, dl-tetrahydrocoptisine and dihydrocoptisine, have been isolated from the methanolic extract of the whole plant of Argemone mexicana L. The compounds were identified by spectral and chemical evidence. This is the first report of these alkaloids in this plant species. PMID- 20013476 TI - A new isoflavone from the fruits of Madhuca latifolia. AB - A new isoflavone, 3',4'-dihydroxy-5,2'-dimethoxy-6,7-methylendioxy isoflavone, was isolated from the Madhuca latifolia fruit coatings. The structure was elucidated on the basis of spectral and chemical evidence. PMID- 20013477 TI - A new anthraquinone derivative from the marine endophytic fungus Fusarium sp. (No. b77). AB - A new anthraquinone compound, 5-acetyl-2-methoxy-1,4,6-trihydroxy-anthraquinone (1), together with four known naphthoquinones (2-5), were isolated from the culture of the mangrove fungus (No. b77). All the isolated compounds (1-5) and the acetyl derivative (1a) were established by comprehensive analysis of the spectral data, especially 1D- and 2D-NMR (HMQC and HMBC) spectra. In the primary bioassays, compound 3 showed strong inhibitory effects on the growth of hepG2 and hep2 cells, with IC(50) values of 1.0 and 2.5 microg mL(-1), respectively. PMID- 20013478 TI - Panajaponin, a new glycosphingolipid from Panax japonicus. AB - Panajaponin, a new glycosphingolipid compound (1), together with eight known compounds, 28-glu-oleanolic acid ester (2), chikusetsusaponin IVa (3), chikusetsusaponin IV (4), ginsenoside Ro (5), ginsenoside Re (6), notoginsenoside R2 (7), ginsenoside Rg2 (8) and adenosine (9), was isolated from Panax japonicus, and the structures of all of the compounds were established on the basis of NMR and MS spectra. PMID- 20013479 TI - Microelement contents and fatty acid compositions of Rhus coriaria L. and Pistacia terebinthus L. fruits spread commonly in the south eastern Anatolia region of Turkey. AB - Sumac (Rhus coriaria L.) and terebinth (Pistacia terebinthus L.) are two important spice plants of south eastern Anatolia, Turkey. Nutrients, physical properties including moisture, ash, dry matter, protein, fatty oil and essential oil content, along with Al, Ca, Cd, Cr, Cu, Fe, K, Mg, Na, Ni, Pb, Zn and characteristics of fruit sizes and fatty acid compositions of both plants were determined from ripe fruits. The free fatty acid content was determined in the fruit oil, and the main fatty acids of sumac and terebinth were identified by gas chromatography/mass spectrometry. They included oleic, linoleic, palmitic and stearic acids in a ratio of 37.7% and 34.8%, 27.4% and 17.3%, 21.1% and 21.7% and, 4.7% and 2.5%, respectively. The fruits of both plants were rich in oil, fatty acids and minerals, suggesting that they are valuable for using in food. The data may also be useful for the evaluation of nutritional information. PMID- 20013480 TI - Dopaminergic modulation of the planning phase of skill acquisition in Parkinson's disease. AB - Patients with Parkinson's disease (PD) are often impaired when performing motor acts and in the acquisition of new motor skills. However, the role of dopamine in developing plans for skill acquisition is unclear. To assess the role of dopamine on the planning of actions, we tested 12 PD and 12 matched normal participants on two skill acquisition tasks matched for motor demands, but varying in requirements for planning. The participants with PD were tested on these tasks when they were on and off dopaminergic medications. To minimize influence of movement related deficits, the subjects used a computer track-pointer that generated the required straight lines when the subjects applied a slight force and clicked the track-pointer to initiate and terminate each line segment. The amount of time the track-pointer was deflected determined the line lengths, while clicking of the mouse determined the location of the line. The simple figure replication task only required the subjects to repeatedly generate lines of two sizes, while the complex figure replication task required subjects to generate lines of different sizes. Thus, this complex task demanded more anticipatory planning. Compared to controls, the subjects with PD were slower to learn the programs needed to produce these figures and produced figures with reduced amplitudes on both the simple and complex tasks. Dopamine treatment, however, only improved the speed of figure completion on the complex task, suggesting that dopamine is important in action planning. PMID- 20013481 TI - On the healthcare question. PMID- 20013482 TI - Response to open peer commentaries on "A duty to deceive: placebos in clinical practice". PMID- 20013483 TI - Response to open peer commentaries on "Journal editorial policies, animal welfare, and the 3Rs". PMID- 20013484 TI - A duty to deceive: placebos in clinical practice. AB - Among medical researchers and clinicians the dominant view is that it is unethical to deceive patients by prescribing a placebo. This opinion is formalized in a recent policy issued by the American Medical Association (AMA [Chicago, IL]). Although placebos can be shown to be always safe, often effective, and sometimes necessary, doctors are now effectively prohibited from using them in clinical practice. I argue that the deceptive administration of placebos is not subject to the same moral objections that face other forms of deception in clinical practice and medical research. Although deception is normally objectionable on the grounds that it limits autonomy and breaches trust, these grounds do not apply to placebos when they are prescribed within appropriate ethical limits. Patients have reason to prefer that doctors can prescribe placebos in ethically responsible ways. Hence, the AMA has an obligation to endorse and to promote the responsible use of deceptive placebos in clinical practice. PMID- 20013485 TI - Mapping community concerns about radical extensions of human life expectancy. PMID- 20013486 TI - Medicine's continuing quest for an excuse to avoid relationships with patients. PMID- 20013487 TI - The primacy of autonomy, honesty, and disclosure--Council on Ethical and Judicial Affairs' placebo opinions. PMID- 20013488 TI - The paradoxical placebo. PMID- 20013489 TI - Paternalistic assumptions and a purported duty to deceive. PMID- 20013490 TI - Physician deception and patient autonomy. PMID- 20013491 TI - Against placebos. PMID- 20013492 TI - How placebo deception can infringe autonomy. PMID- 20013493 TI - The placebo response: the shared construction of reality and the illusion of autonomy. PMID- 20013494 TI - When doctors deceive. PMID- 20013495 TI - When subtle deception turns into an outright lie. PMID- 20013496 TI - Placebos in clinical practice and the power of suggestion. PMID- 20013497 TI - Doctoring the evidence: the case against lying to patients about placebos. PMID- 20013498 TI - Placebo is far from benign: it is disease-mongering. PMID- 20013499 TI - The legitimacy of placebo treatments in clinical practice: evidence and ethics. AB - Physicians commonly recommend 'placebo treatments', which are not believed to have specific efficacy for the patient's condition. Motivations for placebo treatments include complying with patient expectations and promoting a placebo effect. In this article, we focus on two key empirical questions that must be addressed in order to assess the ethical legitimacy of placebo treatments in clinical practice: 1) do placebo treatments have the potential to produce clinically significant benefit? and 2) can placebo treatments be effective in promoting a therapeutic placebo response without the use of deception? We examine evidence from clinical trials and laboratory experiments bearing on these two questions. The conclusion is reached that based on currently available evidence, it is premature to judge whether placebo treatments are ethically justifiable, with the possible exception of acupuncture for pain relief. PMID- 20013500 TI - Placebo prescriptions are missed opportunities for doctor-patient communication. PMID- 20013501 TI - Placebo effects without placebos? More reason to abandon the paradoxical placebo. PMID- 20013502 TI - Is there a place for (deceptive) placebos within clinical practice? PMID- 20013503 TI - Journal editorial policies, animal welfare, and the 3Rs. AB - This study evaluates the editorial policies of a randomized sample of English language peer-reviewed journals that publish original research involving the use of animals. The aim is to identify whether journals have editorial policies relating to the use of animals in the research that they are prepared to publish and whether any policies are likely to promote animal welfare and dissemination of information on the 3Rs (reduction, refinement, replacement) within the scientific community. The results demonstrate that a significant proportion of journals publishing original research involving animals do not have any editorial policy relating to the use of animals. Of those journals that do have policies the majority simply request that the research be carried out in accordance with standard regulatory requirements. This paper aims to provide editors and publishers with the information they need to review their own editorial policies to ensure they are fulfilling their potential to promote animal welfare and dissemination of the 3Rs. PMID- 20013504 TI - Information on the 3Rs in animal research publications is crucial. PMID- 20013505 TI - A question of ethics--publication policy and animals in research. PMID- 20013506 TI - Scientific autonomy and the 3Rs. PMID- 20013507 TI - Underdetermined interests: scientific 'goods' and animal welfare. PMID- 20013508 TI - Can journal editors police animal welfare? Three Es for three Rs in scientific journals. PMID- 20013509 TI - Ethical concerns in the community about technologies to extend human life span. AB - Debates about the ethical and social implications of research that aims to extend human longevity by intervening in the ageing process have paid little attention to the attitudes of members of the general public. In the absence of empirical evidence, conflicting assumptions have been made about likely public attitudes towards life-extension. In light of recent calls for greater public involvement in such discussions, this target article presents findings from focus groups and individual interviews which investigated whether members of the general public identify ethical issues surrounding life-extension, and if so, what these ethical issues are? In this study, while some participants were concerned primarily with the likely personal consequences of life-extension, for others the question of whether or not to pursue interventions to extend longevity, and how they should be implemented, clearly raised important ethical issues, many of which have been prominent in debates among bioethicists. PMID- 20013510 TI - Finding a context for discussing human life-extension. PMID- 20013511 TI - Evaluating life extension from a narrative perspective. PMID- 20013512 TI - The salience of language in probing public attitudes about life extension. PMID- 20013514 TI - The role of working memory in information acquisition and decision making: lessons from the binary prediction task. AB - The effects of memory constraints upon information acquisition and decision making were examined in two experiments using binary prediction tasks, where participants observe outcomes for two options before deciding which one to bet upon. Our studies extend previous investigations to the case where participants learn the structure of the task through observation, but where information acquisition is separated from the task of prediction. Participants with higher cognitive capacity (larger memory span or higher intelligence) were more likely to adopt the "maximizing" strategy (always selecting the more frequent alternative). This finding conflicts with some recent investigations of similar tasks, a contrast that implies that the presence of feedback on choices may be important in determining the strategic actions of high-capacity individuals. Participants selecting the optimal strategy were in turn more efficient in their data acquisition. The behaviour of participants adopting suboptimal choice strategies was consistent with prediction based upon a "narrow window of experience"-that is, seeking to match the characteristics of small samples of observations. PMID- 20013515 TI - Sex differences in the processing of flankers. AB - The study of sex differences in cognition has often focused on differences in spatial processing. Recently, sex differences in selective attention have been observed by Bayliss, di Pellegrino, and Tipper (2005), showing that women are more influenced than men by irrelevant spatial cues. The current study elaborates on this finding and tests whether sex differences in the processing of irrelevant information also occur in a simpler task, in which there is no need to redirect visual attention and no need to remember multiple spatial stimulus-response associations. Here, attention is studied using a novel combination of a go/no-go task and a flanker task. A total of 80 neurotypical participants were studied, and it was found that responses in women were more strongly affected by flanker information than were responses in men. This suggests that these sex differences were not due to difficulties with spatial reorientation, or remembering spatial stimulus-response relationships. The findings are discussed in the context of the hunter-gatherer theory of sex differences. PMID- 20013516 TI - The quality of life of single mothers making the transition from welfare to work. AB - This study examined the quality of life of single mothers making the mandatory transition from welfare to work. The Australian government purported that the benefits of making this transition would include higher incomes, better social participation, and improved wellbeing. It is currently unknown, however, how single mothers currently engaged in welfare to work programs evaluate their quality of life. Quality of life scores for 334 single mothers engaged in welfare to work in Australia were compared with normative data. Participants reported significantly lower quality of life scores than the general population for all quality of life domains, highlighting the need to carefully examine welfare to work policies to ensure they promote participants' quality of life. PMID- 20013517 TI - Predictors of mothers' postpartum body dissatisfaction. AB - PURPOSE: To investigate changes in mothers' body dissatisfaction from delivery to 9 months postpartum, and the relationship of postpartum body dissatisfaction to weight, other health, and social characteristics. METHODS: In this prospective longitudinal study, 506 mothers completed surveys at 0-1 and 9 months postpartum. Postpartum changes in body dissatisfaction and weight were evaluated by paired t tests, and predictors of postpartum body dissatisfaction were identified by stepwise multiple regression analysis. RESULTS: Mothers' body dissatisfaction increased significantly from 0-1 to 9 months postpartum (mean scores of 15.2 and 18.2, respectively, p < .001). Although women lost an average of 10.1 pounds (sd = 16.3) or 4.6 kg. (sd = 7.4) between 0-1 and 9 months postpartum (p < .001), their weight at 9 months postpartum remained an average of 5.4 pounds (sd = 15.6) or 2.5 kg (sd = 7.1) above their pre-pregnancy weights (p < .001). Body dissatisfaction at 9 months postpartum was associated with overeating or poor appetite, higher current weight, worse mental health (SF-36 Mental Health scale), race other than black, bottle-feeding (vs. breastfeeding), being single (vs. married), and having fewer children. CONCLUSIONS: Mothers' body satisfaction worsened from 1 to 9 months postpartum, and 9-month body dissatisfaction was associated with eating/appetite abnormalities, greater weight, worse mental health, non-black race, non-breastfeeding status, and fewer immediate family relationships. Given these relationships, it is important to educate women about expected postpartum weight and body changes, and to find ways to enhance mothers' postpartum self-esteem and body satisfaction. PMID- 20013518 TI - Perspectives of Latina and non-Latina white women on barriers and facilitators to exercise in pregnancy. AB - Exercise during pregnancy has been associated with reduced risk of gestational diabetes mellitus, however, twice as many women are sedentary during pregnancy as compared to when they are not pregnant. We conducted 3 focus groups among 20 pregnant Latina and non-Latina white women to identify barriers and facilitators to exercise in pregnancy to inform a gestational diabetes mellitus intervention study. Quantitative analyses of demographic data, and qualitative analyses of focus groups were conducted. Women identified physical limitations and restrictions, lack of resources, energy, and time as powerful exercise barriers. Social support, access to resources, information, proper diet, scheduling, and the weather were identified as powerful facilitators. Intervention programs designed for pregnant women should facilitate social support, provide information and resources, as well as promote short-term and long-term benefits. PMID- 20013519 TI - Women Bound to be Active: one year follow-up to an innovative pilot intervention to increase physical activity and self-worth in women. AB - The purpose of this investigation was to assess the effectiveness of a lifestyle intervention (a women's book club; Women Bound to be Active) in promoting long term physical activity. Thirty-five women (26-70 years; mean age 50.6 years) completed the 8-month intervention and participated in the one-year follow-up. At follow-up, physical activity returned to baseline levels; however, self-worth and body mass index significantly improved. Women were more knowledgeable about physical activity at follow-up; however, they failed to maintain physical activity after the intervention. Components of the intervention were effective in improving self-worth and lowering BMI at one-year follow-up. To enhance long-term physical activity adherence, continued research and intervention modifications are needed. PMID- 20013520 TI - The experience of group weight loss efforts among lesbians. AB - OBJECTIVE: To describe experiences of group weight loss efforts among lesbians participating in a predominantly lesbian weight loss group. METHODS: A qualitative study (N = 14 self-identified overweight lesbians) was conducted, incorporating phenomenology and grounded theory in methodology. Focus groups were analyzed using a Template Analysis style. RESULTS: Several themes were identified that contributed to the weight loss experience of the participants. These themes contributed to the development of a model depicting history, components, and relationships among concepts leading to positive weight loss experiences for lesbians. CONCLUSIONS: Participants had long histories of shame and fear surrounding weight loss attempts. Weight loss group participants needed cultural connectivity and a sense of safety and acceptance to address issues contributing to weight gain and in order to lose weight and maintain weight loss. Although more research is needed before implementation of a sexual minority women-specific weight loss program, these data are the basis for further exploration into the development of such a program. PMID- 20013521 TI - Experiences of mistreatment among women with cerebral palsy. AB - Abuse and mistreatment of women with disabilities is a complex problem that affects their health and well-being. Previous studies have focused on heterogeneous groups of women with disabilities, with only small numbers of women with cerebral palsy included, but different disabilities may play specific roles in relation to abuse. Exploring mistreatment of women with cerebral palsy is important in determining the relationship between mistreatment and a specific disability. The aim of this article was to describe experiences and meanings of mistreatment among women with cerebral palsy. The feminist biographical method was used to provide an in-depth exploration of women's storied lives, uncover the meaning of women's lives from their own perspective, and provide understanding of women whose stories are seldom told. A sample of eight participants participated in two in-depth, audio-recorded interviews. Two major themes and five subthemes emerged. The meaning of mistreatment included participants' definition of mistreatment and their explanation for mistreatment. Outcomes of mistreatment were divided into emotional, social, and physical outcomes. Health care providers need to understand the meaning and outcomes of mistreatment in their patients' lives to begin to address mistreatment, listen to patients, advocate when needed, and provide appropriate health care. PMID- 20013522 TI - From a declaration of values to the creation of value in global health: a report from Harvard University's Global Health Delivery Project. AB - To make best use of the new dollars available for the treatment of disease in resource-poor settings, global health practice requires a strategic approach that emphasises value for patients. Practitioners and global health academics should seek to identify and elaborate the set of factors that drives value for patients through the detailed study of actual care delivery organisations in multiple settings. Several frameworks can facilitate this study, including the care delivery value chain. We report on our efforts to catalyse the study of health care delivery in resource-limited settings in the hope that this inquiry will lead to insights that can improve the health of the neediest worldwide. PMID- 20013523 TI - Anthropology, knowledge-flows and global health. AB - Global health programmes are damaged by blockages in the upward flow of information from localities and regional centres about realities of professional practice and about patients' lives and conditions of treatment. Power differentials between local actors and national or international decision-makers present further obstacles to effective action. Anthropological research and action, in its most effective current forms, make important contributions to these issues. This research often continues over the long term, intensively. It can be multi-sited, studying actors at local, national and international levels simultaneously. It studies the relative knowledge and power of impoverished patients and global decision-makers, all within a single frame. By doing so, anthropological research is capable of providing new and important insights on the diverse meanings of patient decision-making, informed consent, non compliance, public health reporting, the building of political coalitions for health and many other issues. PMID- 20013525 TI - Coagulopathies and thrombosis: usual and unusual causes and associations, Part II. PMID- 20013526 TI - Unsuspected triggers of venous thromboembolism--trivial or not so trivial? AB - Venous thromboembolism (VTE) can be considered a multifactorial disorder involving a variety of inherited and acquired prothrombotic conditions and events. Although greater emphasis has classically been given to traditional thrombophilic risk factors, there is increasing recognition of less typical precipitating conditions and events. Indeed, the list of plausible but unusual triggers of thrombosis includes sneezing and coughing attacks, eating, migraine, sexual intercourse, strenuous physical exercise, drug abuse, and defecation. Although it is difficult to assert conclusively the true contribution of such events to the etiology of acute episodes of venous thrombosis, it seems reasonable to conclude that the concomitant presence of such trivial elements with one or more additional risk factors for VTE might precipitate an acute thrombotic episode. PMID- 20013527 TI - Hemostatic and fibrinolytic abnormalities in endocrine diseases: a narrative review. AB - This review summarizes current knowledge of the effects of polycystic ovary syndrome, Cushing's syndrome, thyrotoxicosis, hypothyroidism, primary hyperparathyroidism, acromegaly, hypopituitarism, and growth hormone deficiency on coagulation and fibrinolysis. Several abnormalities of the coagulation fibrinolytic system have been described among patients affected by these endocrine disorders. Although further larger studies are needed to provide more definitive information, clinically overt hypothyroidism appears to be associated with a bleeding tendency, whereas all other endocrine diseases appear to be associated with a thrombotic tendency. The disorders of coagulation and fibrinolysis observed in these endocrine pathologies usually range from mild to moderate and, rarely, to potentially severe laboratory abnormalities (e.g., bleeding diathesis in overt hypothyroidism mainly due to an acquired von Willebrand's syndrome type 1), are reversible after pharmacologic treatment of the hormonal dysfunction, and are usually of limited consequence in clinical practice. Nevertheless, the prompt recognition of potentially severe disorders of blood coagulation is mandatory for the correct management of these patients. PMID- 20013528 TI - Polycystic ovarian syndrome and the risk of cardiovascular disease and thrombosis. AB - Polycystic ovary syndrome (PCOS) is a common endocrine disorder associated with multiple comorbidities such as diabetes, dyslipidemia, hypertension, and metabolic syndrome, all of which predispose women with PCOS to early atherosclerosis. Women with PCOS also have a higher prevalence of subclinical atherosclerosis, as reflected in dysregulation of endothelial function, increased carotid intimal-medial thickness, and presence of coronary artery calcification. Preliminary data indicate that serum biomarkers of cardiovascular disease such as high-sensitivity C-reactive protein, homocysteine, and adiponectin are abnormal in women with PCOS. There is limited data on abnormalities in the coagulation and fibrinolytic systems, however. The risk of venous thrombosis is unclear in the PCOS population, and further studies are urgently required to address whether first-line treatment for PCOS with oral contraceptive pills is advisable. PMID- 20013529 TI - Thrombosis in systemic lupus erythematosus. AB - Systemic lupus erythematosus (SLE) is a potentially fatal multiorgan inflammatory disease that primarily affects females. Due to the heterogeneity of clinical manifestations and lack of laboratory tests that are both specific and sensitive for the disease, diagnosis of SLE can often be difficult. Although the precise etiology remains to be fully elucidated, it is probable that various environmental, genetic, and hormonal factors contribute to the development of the disease. Patients with SLE have an increased risk for premature thrombosis and/or atherosclerosis, with up to half experiencing a thrombotic event. Furthermore, antiphospholipid antibodies probably play a key role in the development of thrombosis by affecting various hemostatic protein interactions with phospholipids and cell surfaces as well as platelet function. Despite recent advances in knowledge related to the factors that contribute to the pathophysiology of SLE, numerous challenges related to earlier diagnosis as well as the prediction and prevention of thrombotic events remain to be fully addressed. PMID- 20013530 TI - Role of thrombophilia in adverse obstetric outcomes and their prevention using antithrombotic therapy. AB - A series of case-control studies in the last decade have shown the role of inherited thrombophilia in the occurrence of adverse obstetric outcomes. In small series of cases, it has been proven that rare inherited causes of thrombophilia such as natural anticoagulant deficiencies can be associated with fetal losses. The confirmed presence of antiphospholipid antibodies in plasma, representing an acquired thrombophilic condition, is also an established cause of fetal losses, although other studies with a smaller sample size have found an association with other obstetric complications, namely preeclampsia, fetal growth restriction, and abruption placentae. Case-control studies have been performed regarding the potential association between unexplained fetal losses and mild hyperhomocysteinemia. Although case-control and prospective studies are also available regarding hyperhomocysteinemia and other gestational vascular complications, published data are conflicting. Intervention studies have been performed to prevent adverse obstetric outcomes in women with inherited or acquired thrombophilia and previous adverse outcomes. There is much debate in the literature regarding the need for treatment of women with thrombophilia during pregnancy. Although in most cases these are not randomized controlled trials, all studies found significantly better outcomes in treated pregnancies compared with those of untreated pregnancies. PMID- 20013531 TI - The bidirectional relationship of cancer and hemostasis and the potential role of anticoagulant therapy in moderating thrombosis and cancer spread. AB - The relationship between hemostasis and malignancy is well recognized, with both elements interacting in a "vicious cycle" where cancers overexpress procoagulants and thrombin, which in turn promote both prothrombotic potential and tumor growth, invasion, and spread. Indeed, venous thromboembolism, particularly idiopathic venous thrombosis, occurs frequently as a paraneoplastic phenomenon, and in turn several components of primary and secondary hemostasis (namely platelets, tissue factor, and thrombin) play an important role in primary tumor growth and metastasization. Despite the many and various mechanisms involved in this multifaceted relationship, anticoagulants might represent an attractive anticancer therapy, in that current research supports the hypothesis that such drugs may offer a better control of cancer progression. The main biological and clinical evidence on the relationship between cancer and hemostasis are briefly summarized in this review, as is the potential benefits of anticoagulant therapy in this setting. PMID- 20013532 TI - Prostate-specific antigen, prostate cancer, and disorders of hemostasis. AB - Prostate cancer is the most prevalent malignancy in men and the third leading cause of cancer deaths worldwide. Disorders of hemostasis are commonplace in patients with prostate cancer and include disseminated intravascular coagulation, venous thromboembolism, acute coronary syndrome, and postsurgical bleeding. These hemostatic disorders contribute to the mortality and morbidity of prostate cancer. The leading mechanisms proposed to underlie prostate cancer-related coagulopathies are thought to be a hyperexpression of tissue factor, cancer procoagulant, and platelet-activating factor, which is then accompanied by release of large amounts of both prothrombotic and profibrinolytic substances into the bloodstream. Given the generally accepted notion that prostate-specific antigen (PSA) represents an important biomarker in prostate cancer diagnostics, large population screenings were initiated for early detection of cancer. However, recent clinical and economic drawbacks have been recently raised, including evidence that screening exposes patients to a significant risk of both overdiagnosis and overtreatment. Nevertheless, several lines of evidence suggest that PSA may have tumor-suppressing activities. Despite being a member of the vast kallikrein family, which actively interplays with the coagulation cascade, the role of PSA in the pathogenesis of hemostatic disorders observed in prostate cancer patients remains circumstantial and speculative. However, observations that the levels of this cancer marker tend to correlate positively with those of several markers of thrombin generation, and with postsurgical bleeding as well as with coronary atherosclerosis and negative outcomes of myocardial infarction, raise a new and intriguing scenario regarding the pathophysiological role of this serine protease. PMID- 20013533 TI - Particulate air pollution and cardiovascular risk: short-term and long-term effects. AB - It is well known that exposure to particulate air pollutants is associated with the development of adverse health effects. In particular, there is growing evidence of a close relationship between increased levels of environmental pollutants and cardiovascular disease. Particle exposure may trigger acute cardiac events as well as promote the chronic development of cardiovascular disorders. Several biological mechanisms have been suggested to explain the important effect of air pollution on cardiovascular morbidity and mortality, although the exact pathways are not fully understood. The literature data on the short-term and long-term cardiovascular effects of particulate air pollutants are discussed in this review from a clinical and mechanistic point of view. PMID- 20013534 TI - Genetic architecture of coronary artery disease in the genome-wide era: implications for the emerging "golden dozen" loci. AB - Clinicians are well aware of family history as a risk factor for coronary artery disease (CAD) and myocardial infarction (MI). The underlying genetic architecture of CAD/MI is extremely complex and still poorly understood. Overall, the genetic heritability of CAD/MI is estimated to be near 40 to 60%. This proportion includes mainly genes that regulate known risk factors (e.g., lipid metabolism) but also genes involved in as yet unknown metabolic pathways. In the last 2 years, the systematic application of genome-wide association studies in the setting of large collaborative consortia including thousands of patients and controls has led to the identification of several new loci associated with CAD/MI. Here we review current knowledge on the emerging "top" 12 loci, that is, those showing the most consistent associations with clinical phenotypes. Although these genetic variants have little or no current predictive value of at the level of individual patients, they have the potential to disclose novel biological mechanisms involved in the pathophysiology of CAD/MI. PMID- 20013536 TI - Laboratory investigation of thrombophilia: the good, the bad, and the ugly. AB - Thrombophilia can be broadly defined as an increased tendency toward hypercoagulability and venous thrombosis. There are several defined risk factors for thrombosis, and these are generally distinguished as either acquired or congenital, although sometimes this distinction is blurred because of interrelationships. Congenital risk factors include deficiencies or defects in natural anticoagulants, such as antithrombin, Protein C and Protein S, and genetic polymorphisms such as prothrombin G20210A and cleavage-resistant forms of factor V (in particular factor V Leiden), that lead to a condition commonly known as activated protein C resistance. Acquired risk factors include antiphospholipid antibodies, detected as lupus anticoagulants and/or anticardiolipin antibodies and/or anti-beta-2-glycoprotein-I antibodies. High levels of clotting factors, dysfibrinogenemia, hyperhomocysteinemia, prolonged immobilization, increasing age, surgery, trauma, cancer, obesity, poor nutrition, pregnancy, oral contraceptives, and hormone replacement therapy comprise just some of the other risk factors. Each of these elements constitutes a component of increased risk, which is compounded when concomitant. There is ongoing debate regarding relative and compound risks, the value of laboratory screening, whom and when to screen for these markers, which tests and methodologies to use, and the form and duration of therapeutic management. The current article explores several important issues primarily from a scientific perspective and predominantly related to laboratory testing. Many of these issues appear to be simply overlooked by some clinicians managing patients with thromboses. In brief, although there is potential significance in testing for various thrombophilia associated markers, this value is limited and greatly diminishes when inappropriately applied. The application of excessive or inappropriate thrombophilia testing is of particular concern, and the net effect of current worldwide testing trends is likely to be more detrimental than beneficial. In short, it is likely that current generalized testing is simply doing more harm than good, and thus that ordering practice requires scrutiny. PMID- 20013535 TI - Inherited thrombophilia: implications for prevention and treatment of venous thromboembolism. AB - Inherited thrombophilia, defined as a genetically determined tendency to develop venous thromboembolism (VTE), contributes to the pathogenesis of approximately 40% of VTE episodes. About 50% of carriers of inherited thrombophilic traits develop VTE, but the impact of the different abnormalities is variable in terms of clinical penetrance. Some rare abnormalities (natural anticoagulant deficiencies, homozygous factor V Leiden, and combined defects) result in more severe thrombophilic phenotypes, characterized by early-onset events, more frequent recurrence, and positive family history, whereas the common polymorphisms (heterozygous factor V Leiden and prothrombin G20210A) are associated with lower VTE risk, often in association with triggering risk factors. Therefore, clinical implications of inherited thrombophilia should be assessed in the framework of coexisting and/or circumstantial risk factors involved in the multifactorial pathogenesis of VTE. These considerations should be taken into account when assessing the need and modalities of primary and secondary thromboprophylaxis in patients carrying inherited thrombophilic traits. PMID- 20013538 TI - Inhaled BDP/formoterol extra-fine combination. Evidence and future perspectives. AB - The combination of inhaled corticosteroids (ICS) and long acting Beta-2 agonists (LABA) represents the mainstay of current treatment of moderate to severe persistent asthma. Corticosteroids and LABA combine the main pillars of asthma therapy--inhibition of inflammation and bronchodilation--and they may potentiate each other when they simultaneously reach the pharmacological target. A new extra fine formulation containing the combination of inhaled beclometasone (BDP) and formoterol (F) (with the non polluting HFA-Hydrofluoroalkane-134a as propellant) is now available on the market. The extra-fine formulation increases the deposition into the peripheral airways and a greater proportion of the inhaled compound reaches the pharmacological target. Thus, the dose of ICS can be decreased and the risk-benefit profile improves. The efficacy and tolerance of BDP/F extra-fine combination has been documented in randomized clinical trials into which patients with moderate to severe asthma were included. These trials confirmed that the new extra-fine combination is as effective as the other fixed combinations. PMID- 20013539 TI - [COPD: from obstructive lung disease to chronic systemic inflammatory syndrome?]. AB - COPD is a complex and heterogeneous disease, in which multiple genes and environmental factors are involved. The disease is associated with a high risk of comorbidity and systemic manifestations. The majority of the patients with COPD suffer from additional chronic diseases like hypertension, diabetes, heart disease, osteoporosis or cancer. Parameter, assessing selectively the lung function, are only conditionally suited for capturing the COPD as a whole. In future subtyping of COPD might give the opportunity to better understand the heterogeneity of the disease. A holistic approach, understanding COPD not as an isolated disease of the lung, but as a "chronic systemic inflammatory syndrome" (CSIS), promises further improvement. Systemic inflammations seems to be a common mechanism leading to chronic diseases by sharing risk factors like smoking, overweight, hypertension and elevated blood lipids. In response to this approach, the diagnosis and therapy of COPD must include the whole entity of comorbidities. PMID- 20013541 TI - [Treatment of small airway diseases--today and tomorrow]. PMID- 20013540 TI - [Inhalative pharmacotherapy in the future--nanocarriers for pulmonary drug delivery]. AB - Nanotechnological approaches promise considerable potential for the treatment of pneumological diseases, especially asthma and COPD (Chronic Obstructive Pulmonary Disease). At present numerous nanotechnological approaches are under examination, which allow a controlled release of encapsulated drugs or diagnostics. These include nanocarriers composed of polymers or anorganic materials, liposomes, micelles, quantum dots, dendrimeres, polymer conjugates and other systems. Polymeric nanocarriers give the opportunity to control the temporal as well as the spatial distribution of drugs within the lung. By varying the composition, structure and dimension nanoparticles could offer a controlled and prolonged duration of effect of the encapsulated drugs as well as a regional and cell specific drug targeting within the lung. PMID- 20013542 TI - [Asthma--a small airway disease: concepts and evidence]. AB - Inflammatoric processes in the large airways have traditionally been considered to be very important. But as new studies show inflammation and structural alteration also take place in the peripheral airways. Meanwhile it is highly evident that asthma is related to all areas of the lung--to the large and even the small airways. But the inflammation of the small airways is not sufficiently diagnostically detected. FEV(1) and PEF reflect the participation of the small airways only partially. New findings about inflammation and remodelling in the small airways underline the need of therapies reaching all parts of the lung. A combined therapy with ICS and LABA seem to have the capability to improve efficacy and safety, especially when very fine formulations are chosen. PMID- 20013543 TI - [Music and health--what kind of music is helpful for whom? What music not?]. AB - It is well known that music not only may improve quality of life (QoL) but also have different effects on heart rate (HR) and its variability (HRV). Music emphasis and rhythmic phrases are tracked consistently by physiological variables. Autonomic responses are synchronized with music, which might therefore convey emotions through autonomic arousal during crescendos or rhythmic phrases. A greater modulation of HR, HRV and modulations in cardiac autonomic nerve activity was revealed with a greater effect for music performance than music perception. Reactions to music are considered subjective, but studies suggested that cardiorespiratory variables are influenced under different circumstances. It has been shown that relaxing music decreases significantly the level of anxiety in a preoperative setting to a greater extent than orally administered midazolam (p < 0,001). Higher effectiveness and absence of apparent adverse effects make preoperative relaxing music a useful alternative to midazolam for premedication. In addition, there is sufficient practical evidence of stress reduction to suggest that a proposed regimen of listening to music while resting in bed after open heart surgery. Music intervention should be offered as an integral part of the multimodal regime administered to the patients that have undergone cardiovascular surgery. It is a supportive source that increases relaxation. Music is also effective in under conditions and music can be utilized as an effective intervention for patients with depressive symptoms, geriatrics and in pain, intensive care or palliative medicine. However, careful selected music that incorporates a patient's own preferences may offer an effective method to reduce anxiety and to improve quality of life. The most benefit on health is visible in classic music, meditation music whereas heavy metal music or technosounds are even ineffective or dangerous and will lead to stress and/or life threatening arrhythmias. There are many composers most effectively to improve QoL, particularly Bach, Mozart and Italian composers are "ideal". PMID- 20013544 TI - [Opium in 19th century medicine and literature]. PMID- 20013545 TI - [Medicine and greek tragedy]. PMID- 20013546 TI - [Fever, carbuncles and hollow bones--disease and medicine in Shakespeare's plays]. PMID- 20013547 TI - [The diseases of King Richard III. according to their descriptions in Shakespeare's works]. AB - In this short essay we will discuss the possible diseases of king Richard III according to the descriptions in Shakespeare's plays King Richard III and Henry VI. Furthermore it is shown that the description of the defeated enemy as physically and mentally deformed is part of a long tradition which has its roots in ancient Greece. PMID- 20013548 TI - [Schiller--writer and physician]. PMID- 20013549 TI - [How the physician's finger became a goldfinger]. PMID- 20013550 TI - [A wonderful cure--a short story by Johann Peter Hebels]. PMID- 20013551 TI - The growth hormone receptor (GHR) polymorphism in growth-retarded children with Cushing disease: lack of association with growth and measures of the somatotropic axis. AB - Pediatric Cushing disease (CD) often presents with short stature, but we have observed significant inter-individual variability in the growth delay caused by endogenous hypercortisolism. Glucocorticoids cause growth retardation by affecting the growth hormone (GH) - insulin-like growth factor-1 (IGF 1) somatotropic axis, but also other, GH-independent sites. Recently, the GH receptor (GHR) gene was found to have a common polymorphism (P) that leads to a deletion (d3) or retention of exon 3. In this study, we tested the hypothesis that the GH receptor polymorphism (GHR-P) maybe one of the significant variants that determines the degree of growth delay among patients with CD. GHR genotyping was performed on 56 children with newly diagnosed CD (24 females, 32 males, mean age of 12.9+/-3.3 years) who were followed at our institution between the years 1997-2007. Correlation analysis included genotype, measures of growth and the somatotropic axis, and anthropometrics. Within the group, 31 (12 girls, 19 boys) expressed the full length GHR allele, 10 (4 girls, 6 boys) were d3-GHR homozygotes and 15 (7 girls, 8 boys) were d3-GHR heterozygotes. No significant differences were found between the GHR genotypes and patient's height and/or growth velocity, or any other measures that we evaluated. The presence of a well studied and common GHR polymorphism does not appear to be responsible for the variability of growth delay observed in patients with Cushing disease. PMID- 20013552 TI - Cardiac effects of vagus nerve stimulation in patients with major depression. AB - INTRODUCTION: Changes in the heart rate variability are well known among patients with depression. Amongst others, a modulation of the autonomic nervous system is discussed. An investigation of heart rate variability during terms of stimulation could give some insight in the central nervous effect of vagus nerve stimulation (VNS) and possible cardiac side effects. METHODS: The effects of VNS on heart rate (HR) and heart rate variability were studied (HRV) during stimulation in nine patients with major depression according to ICD-10. RESULTS: When comparing treated depressive patients with a sex- and age-matched healthy control group, the analysis of heart rate revealed significantly (t=2.8; df=16, p=0.012) elevated heart rates during completely switched off conditions, during switched on VNS programme in stimulation-free intervals (5 min period) (t=3.0; p=0.009) and during stimulation conditions (30 s) (t=2.8, Levene corrected, p=0.015). The RMSSD (root mean square of successive differences) as a measure of the HRV increased significantly in switched on conditions during stimulation (30 s) in six patients compared to stimulation-free intervals (t=-4.7; df=5, p=0.006) and baseline. DISCUSSION: Clinically relevant cardiac effects were not observed throughout the study. VNS induces reversible changes in heart rate variability in patients with major depression during stimulation conditions. PMID- 20013553 TI - Prescribing a sedative antidepressant for patients at work or on sick leave under conditions of routine care. AB - BACKGROUND: Sedation can be a beneficial effect of medication, but it can also be an unwanted side-effect, especially in patients who have to work. The aim of this study is to analyze whether physicians prescribe sedative antidepressants differently for patients at work vs. those on sick leave. METHODS: A drug utilization study of mirtazapine was conducted for 12 weeks on 594 depressed outpatients from 227 general practitioners or psychiatrists. There were 319 patients working and 275 patients on sick leave. The two groups were compared regarding sociodemographic variables, illness characteristics, mode and course of treatment. RESULTS: As expected, patients on sick leave were sicker than working patients; they were treated by specialists more often and received higher dosages of mirtazapine. Work status had no influence on dosage after controlling for severity of illness, patient sex, and physician specialty. The overall improvement of depression was similar in both patient groups. Under treatment with mirtazapine, 64.5% of patients on sick leave returned to work, while 2.6% of the patients initially at work went on sick leave by the end of the 12 weeks. Sedation as an undesired side-effect was reported in less than 1%. The rate of sedative polypharmacy declined during treatment with mirtazapine. CONCLUSION: The results suggest that in routine treatment physicians do not see a need to adjust prescribing of mirtazapine because of its sedative properties to the working status of the patients. The majority of patients on sick leave returned to work. Mirtazapine can help to reduce sedative polypharmacy. PMID- 20013554 TI - Clozapine intoxication in a patient with lamotrigine-induced rash. PMID- 20013555 TI - Long-term effects of oxidative stress in volleyball players. AB - The aim of this study was to determine the impact of long-term training on elite female volleyball players and to determine parameters that could discriminate them according to the level of oxidative stress-associated adaptation. Fifty-four elite female volleyball players were divided into 3 groups (1: below-average training experience <8.0 years, 2: average training experience between 8.0 and 10.5 years and 3: above-average training experience >10.5 years). The measured parameters were reactive oxygen metabolites, biological anti-oxidative potential, superoxide anion, malondialdehyde, advanced oxidation protein products, lipid hydroperoxides, paraoxonase activity, superoxide-dismutase activity and sulphydryl groups. Multiple discriminant analysis of the oxidative stress status parameters between the three groups of athletes indicated a statistically significant difference (Wilks' lambda=0.458, X(2)=35.898, p=0.031). The most important discriminant variables, superoxide-dismutase and superoxide anion, were the best indicators of differences between groups with different training experience. The significantly higher values were found in Group 3 compared with Group 1 in superoxide-dismutase activity (141+/-32 vs. 86+/-46; p=0.002), sulphydryl groups (p=0.031), and reactive oxygen metabolites (p=0.042). The significantly lower superoxide anion was found between Group 3 and Group 1 (377+/ 187 vs. 1183+/-905; p=0.001). Oxidative stress status parameters adequately discriminated 68.5% of athletes with different training experience. PMID- 20013556 TI - Shoulder rotators electro-mechanical properties change with intensive volleyball practice: a pilot study. AB - This pilot study was designed to assess the incidence of high-level volleyball practice on muscle strength production and muscle activation during internal and external shoulder rotations. Seven professional and seven French amateur league volleyball players performed maximal isometric at three forearm angles, concentric and eccentric isokinetic internal and external shoulder rotations. The torque production and muscle activation levels of PECTORALIS MAJOR and INFRASPINATUS were determined. Few significant differences were found for muscle activation and co-activation between amateur and professional volleyball players during both internal and external rotations. No significant difference in torque production was observed for shoulder internal rotation between professional and amateur volleyball players. Torque production was significantly higher during shoulder external rotation for amateur (46.58+/-2.62 N . m) compared to professional (35.35+/-1.17 N . m) volleyball players relative to isometric contractions, but it was not different during isokinetic efforts. The torque ratios for external/internal rotations were always significantly lower for professional (0.42+/-0.03 pooling isometric and concentric conditions) compared to amateur volleyball players (0.56+/-0.03 pooling isometric and concentric conditions). Those results emphasize that a high level of volleyball practice induces a strong external rotators deficit compared to sports such as swimming, baseball or tennis. PMID- 20013558 TI - Is there an ACE ID - ACTN3 R577X polymorphisms interaction that influences sprint performance? AB - Functional R577X (rs.1815739) and ID (rs.5186) polymorphisms in the alpha-actinin 3 ( ACTN3) and the angiotensin converting enzyme (ACE) genes, respectively, have been associated with sprint performance. The aim of this study was to determine their effect on sprint performance among 81 Israeli sprinters and 240 healthy controls. Results revealed that the ACE II genotype+ ACTN3 R allele (P=0.003 for sprinters vs. controls), and the ACTN3 RR genotype +ACE I allele (P=0.001 for sprinters vs. controls) might be the genotype for sprinters. In the whole cohort the probability of ACTN3 RR genotype+ ACE I allele being a sprinter (odds ratio 2.67, 95% confidence interval 1.45-4.93) and of ACE II genotype+ ACTN3 R allele being a sprinter (odds ratio 3.57, 95% confidence interval 1.78-7.15) was significantly higher than that in the controls. In conclusion, the above data suggest that ACE ID/ ACTN3 R577X genotype combination is associated with sprint ability. However, ACE ID/ ACTN3 R577X genotype combination is not related to the level of performance. PMID- 20013559 TI - Effect of glycemic index meals on recovery and subsequent endurance capacity. AB - This study examined the effect of ingesting a high or low glycemic index (GI) meal during a short-term recovery period on endurance running capacity. On two occasions, seven men (age 30.0+/-2.6 yr, body mass 60.7+/-1.4 kg, VO (2max) 62.1+/-2.2 ml.kg (-1).min (-1)) ran at 70% VO (2max) on a level treadmill for 90 min (R1), followed by a 4 h recovery (REC) and a further exhaustive run at the same speed (R2). Twenty minutes after R1, each subject consumed an isoenergetic meal containing either high GI (HGI, GI=77) or low GI (LGI, GI=37) carbohydrate providing 1.5 g CHO.kg (-1) BM. During REC, subjects also ingested a prescribed volume of water equal to 150% of their BM loss during R1. The duration of R2 in the HGI trial was 15% longer than in the LGI trial (HGI: 86.6+/-10.7 min vs. LGI: 75.2+/-8.1 min, p<0.05). The subjects also achieved complete rehydration after REC on both occasions. In conclusion, the consumption of a HGI meal during a 4 h recovery improved endurance capacity in a subsequent run; however, the precise mechanism(s) by which this takes place is yet to be clarified. PMID- 20013561 TI - [Klinische Padiatrie and its development during the present decade]. PMID- 20013562 TI - [Morbus Wilson - hepatic manifestation]. PMID- 20013563 TI - [Legal aspects of ritual circumcision]. AB - Female circumcision (genital mutilation) is a criminal violation of human rights under German law. Even with consent of the person to be circumcised and/or her legal representative this procedure must not be carried out since a consent to female circumcision is unethical and therefore void. As much consent as there is on female circumcision the legal situation with ritual male circumcision is very unclear. In practice and unnoticed by the public male circumcision is carried out - be it for medical or ritual reasons - without deeper-going reflexions on the clearness of the medical indication or the legal situation with ritual circumcision. From the medical aspect there are big differences between female and male circumcision but also certain parallels. Various reasons, partly founded in prejudice and misinformation, make people refrain from regarding circumcision of boys also as illegal. Contrary to the prevailing opinion male circumcision also represents a bodily harm which a doctor can only carry out after a preoperative interview and with the consent of the affected person. Since ritual male circumcision does not serve the wellbeing of a child it is not possible for the parents to give their consent to the circumcision in lieu of the child. Male circumcision is only permitted if the child has given his consent and is thus only legally permitted if the child has reached an age at which he is mature enough to understand the meaning and extent of such an action which is hardly the case before he has completed his 16 (th) year. PMID- 20013564 TI - Wilson's disease: monocentric experiences over a period of 10 years [corrected]. AB - BACKGROUND: Wilson's disease (WD) is an autosomal recessive disorder of copper metabolism. The objective of this study is to present diagnostic pitfalls and long time follow-up data in Wilson disease. PATIENTS/METHODS: We studied 21 WD patients and 14 heterozygote carriers aged 2-43 years, retrospectively. 18 WD patients presented liver disease, three had mixed neurological and hepatic involvement and 9 patients underwent orthotopic liver transplantation (OLT). RESULTS: The median age at diagnosis of WD children without OLT was 10.16+/-3.8 (range, 5-16). All of females and younger age categories of patients prevailed in acute liver failure group. Serum ceruloplasmine levels were below 0.2 g/l in about (1/3) of WD carriers (X =0.27+/-0.09 g/l) and nearly (2/3) of children with WD (X = 0.21+/-0.13 g/l). A statistically significant difference (p<0.05) in the 24-h excretion of copper in urine was noticed between healthy controls, children with WD and WD heterozygote carriers. As diagnostic important proved the copper content of more than 250 microg/g hepatic dry weight. The Kayser-Fleischer?s ring was not observed in children. Ceruloplasmine, haemoglobin, ALT, ALP and plasma albumin were significantly different between fulminant and non-fulminant WD and could be used as indirect markers in evaluation of urgent OLT. CONCLUSION: Detection of WD in children remains very difficult. The most important investigation is liver biopsy with the assessment of liver copper. Genetic analysis may help in doubtful cases. PMID- 20013565 TI - Experience with mycophenolate mofetil as maintenance therapy in five pediatric patients with severe systemic lupus erythematosus. AB - Maintenance therapy of severe pediatric systemic lupus erythematosus (SLE) usually consists of azathioprine and prednisone . In adult SLE patients mycophenolate mofetil (MMF) is successfully used, superiority to azathioprine has not been shown yet. We hypothesized that a maintenance therapy with MMF is able to decrease disease activity as well as the dose of glucocorticoid needed in children and adolescents with SLE. Five girls with a mean age of 13.9 (range 12 15) years were treated with 1.2+/-0.20 g/m (2) MMF daily on individual medical decision. Three patients had severe renal (WHO IV) and one severe cerebral involvement. Three patients with frequent flares on azathioprine maintenance therapy were switched to MMF, two patients with severe renal and cerebral manifestation received MMF additionally after induction therapy. Flares, steroid dosage, and disease activity (SLEDAI) were monthly registered in all patients. The number of flares decreased from 1.28 to 0.25 episodes per patient year during a mean follow-up period of 39 (range 36-42) months after MMF initiation. In parallel prednisone dose could be reduced from 10.80+/-5.25 to 3.25+1.18 mg/d (p<0.01). SLEDAI score dropped from 15.20+/-2.8 before MMF to 3.60+/- 0.9 at the last visit under MMF (p<0.001). No severe adverse event occurred. In our cohort of five pediatric patients MMF was effective and safe for maintenance therapy of SLE over a period of 3.5 years. MMF seems to be successful in preventing flares even in adolescents having unfavorable course on azathioprine treatment before. This observation should be confirmed by a randomized multicenter clinical trial. PMID- 20013566 TI - ['Cases against KiSS': a diagnostic algorithm for children with torticollis]. AB - In 1991, Biedermann coined the term "kinetic imbalance due to suboccipital strain" ("KiSS-syndrome"). He assumed a functional abnormality of the suboccipital-high cervical spine, resulting in positional preference of the infant;s head. A broad spectrum of symptoms and complaints have been attributed to "KiSS-Syndrome". Patients are advised to undergo manual therapy, with pressure applied locally in order to readjust the cervical spine. Life threatening side effects have been published repeatedly. We present two infants with brain tumours who developed torticollis and further neurological findings such as ataxia and reflex differences. In both cases, symptoms caused by the tumour were interpreted as "KiSS-syndrome", and appropriate diagnostics and therapy were delayed for months. There is no scientific evidence for the actual existence of "KiSS syndrome" as a clinical entity or for the positive effects of manual therapy. Approximately 12% of all infants <12 months show a positional preference of the head, about 8% present with body asymmetry. Whereas most cases are benign, there is a long list of serious differential diagnoses for torticollis in infants. We give an updated review of the literature regarding "KiSS-Syndrome" and discuss the differential diagnostics in infants with torticollis. PMID- 20013567 TI - Mechanographic analyses in pediatrics: allometric scaling of 'peak jump force' and its relationship to 'maximal isometric grip force' in childhood and adolescence. AB - BACKGROUND: Kinetic analyses of the neuromuscular system have become an important part in the diagnostics of metabolic bone disorders in pediatrics because of the relevance of the 'Functional Muscle-Bone Unit'. Because muscle function is associated with muscle metabolism, it is suggested that the association of maximal forces with anthropometric variables (e. g., body weight) is characterized by allometric scaling. The present manuscript aims to describe the scaling exponent for the association of maximal forces of mechanography and dynamometry to body weight and exemplarily applies the prediction of 'Peak Jump Force' (PJF) by 'Maximal Isometric Grip Force' (MIGF) for the characterization of the unconditioned neuromuscular system in subjects with congenital heart disease (CHD). SUBJECTS: MIGF was measured in a reference population of 135 school boys and 177 school girls who were not randomized individuals of a German primary and high-school. In addition, data of the neuromuscular system were analyzed in 29 individuals with CHD. METHODS: Participants performed counter-movement jumping on a force plate to measure PJF and v (max). MIGF was determined by dynamometric measurement. RESULTS: PJF scaled (body mass) (0.97) in girls and (body mass) (1.05) in boys. PJF was not different from the scaling exponent=1 in both genders. After having taken the logarithm, MIGF predicted PJF with R (2)(adjusted)=0.774 in boys, R (2)(adjusted)=0.720 in girls (p<0.001 each) and R (2)(adjusted)=0.209 (p=0.007) in patients with CHD. The mean of the predicted PJF was lower than the mean of the measured PJF in subjects with CHD (p<0.001). The difference between the predicted and the measured PJF (DeltaPJF) was associated with v (max) (R (2)(adjusted)=0.113, p=0.042). CONCLUSIONS: PJF scales body weight in an isometric way as recently describe for other vertebrates. MIGF is a good predictor of PJF in healthy children, but not in unconditioned individuals. DeltaPJF indicates conditioning of the individual and may be used to describe inter- and intramuscular coordination in children. HINTERGRUND: Mechanografische Analysen des neuromuskularen Systems haben mittlerweile auch einen diagnostischen Stellenwert in der Padiatrie erlangt. Ziel der vorliegenden Untersuchung ist die kinetische Analyse des Counter-movement-Sprungs durch die Messung der Bodenreaktionskrafte in Hinblick auf seine allometrische Skalierung und im Vergleich zur Entwicklung der ,Maximalen isometrischen Griffstarke' (MIGF) bei Kindern und Jugendlichen. PROBANDEN: Die Studienpopulation umfasste 135 Schuler und 177 Schulerinnen, die nicht randomisiert wurden und Schuler einer deutschen Grundschule und eines deutschen Gymnasiums waren. Zudem wurden Daten von 29 Patienten mit kongenitalen Herzfehlern analysiert. METHODEN: Die Studienteilnehmer absolvierten Counter-movement-Sprunge auf einer Sprungplatte, sodass die maximale Sprungkraft (Peak Jump Force, PJF) und die maximale Geschwindigkeit des Massenschwerpunktes beim Absprung (v (max)) ermittelt werden konnten. MIGF wurde durch Messung mit einem Dynamometer bestimmt. ERGEBNISSE: PJF skalierte (Korpermasse) (0,97) bei den Madchen und (Korpermasse) (1,05) bei den Jungen. Der Skalierungsfaktor von PJF war bei Madchen und Jungen nicht signifikant vom Faktor=1 verschieden. Nach Lograithmierung sagte die Variable MIGF den Parameter PJF in der Gruppe der Jungen mit R (2)(adjusted)=0,774, in der Gruppe der Madchen mit R (2)(adjusted)=0,720 (jeweils p<0,001) und in der Gruppe der Herzkranken mit R (2)(adjusted)=0,209 (p=0,007) voraus. Der Mittelwert des vorhergesagten Wertes fur PJF war niedriger als der Wert der gemessenen PJF in Patienten mit Herzerkrankung (p<0,001). Die Differenz zwischen vorhergesagter und gemessener PJF (DeltaPJF) stand statistisch mit v (max) (R (2)(adjusted)=0,113; p=0,042) in Zusammenhang. SCHLUSSFOLGERUNGEN: PJF skaliert isometrisch zur Korpermasse, wie es bereits fur andere Vertebraten beschrieben wurde. MIGF ist ein guter Pradiktor fur PJF bei gesunden Kindern und Jugendlichen, jedoch nicht bei Menschen mit einem unkonditionierten neuromuskularen System. DeltaPJF kann als Marker fur die Konditionierung eines neuromuskularen Systems angesehen werden und beschreibt inter- und intramuskulare Koordination. PMID- 20013568 TI - [Primary ciliary dyskinesia causing neonatal respiratory distress]. AB - BACKGROUND: Primary ciliary dyskinesia (PCD) is a hereditary disorder of structure and function of the cilia of respiratory epithelium of the upper and lower airways. Prevalence is estimated with 1:15 000 to 1:30 000 births. We present a newborn infant with respiratory distress caused by PCD. PATIENT: On the first day of life, the male newborn developed dyspnoe and cyanosis, so that CPAP and short term ventilation was necessary. Varying atelectasis impressed on the chest radiographs and the diagnosis of PCD was made by nasal brush biopsies. Causative is a lack of the inner dynein arms of the cilia. The clinical features of newborns with the diagnoses of PCD are listed and compared with the own case. CONCLUSION: PCD is a rare cause of neonatal respiratory distress and should be considered in term infants with unknown and prolonged course even if Situs inversus is lacking. PMID- 20013569 TI - Cardiogenic shock in a neonate with enterovirus myocarditis. AB - BACKGROUND: Left ventricular compromise in the neonate may be caused by birth asphyxia, metabolic disease, congenital heart disease, and systemic bacterial or viral infections. In rare cases, enterovirus infection may cause severe disease including cardiac, cerebral, hepatic and multi organ failure. PATIENTS AND METHODS: Case report. RESULTS: A 3-week-old neonate was admitted to our NICU in cardiogenic shock and severe lactic acidosis (ph: 6.9; serum lactate: 15 mmol/l, base excess: -19.8 mmol/l; pCO (2): 54.9 mm Hg). Serum troponin T was within the normal range; serum total creatinin phosphokinase was 57 U/l, CK-MB 110 U/l, LDH 762 U/l; pro-BNP: 64391 pg/ml was elevated. On echocardiography left ventricular function was depressed with a shortening fraction of 16%. The neonate was started on inotropes. There was gradual improvement over the following two weeks with normalisation of left ventricular output. PCR examination was positive for enterovirus. Other causes for left ventricular compromise (congenital heart disease, inborn errors of metabolism, etc.) were ruled out by adequate means. CONCLUSIONS: Enterovirus infection as a cause for myocarditis and cardiogenic shock should be taken into the differential diagnosis in neonates. PMID- 20013570 TI - Oral Valganciclovir treatment in a CMV congenital infected infant with sensorineural hearing loss (SNHL) first detected at 4 months of age. PMID- 20013571 TI - [Developmental screening of preterms at an early stage postpartum]. AB - Preterm infants are at high risk to develop neurological disorders. Therefore extensive follow-up programs with a combination of various instruments are needed for early detection of infants with major or minor developmental disorders and behavioral problems. The Newborn Individualized Developmental Care and Assessment Program (NIDCAP) is based on standardized observation protocols and provides a powerful method to assess behavior and brain development directly after birth, so can be used as an early screening instrument. PMID- 20013572 TI - [Evidence-based treatments in the rehabilitation of patients with depression--a literature review]. AB - BACKGROUND: In recent years, the importance of guidelines has increased continuously. This development also occurs in the field of rehabilitative health care, where process guidelines are being designed for various indicational groups to ensure quality standards and improvements. AIM: The primary goal of this paper is to collect and evaluate the evidence for various treatment options for depressive disorders in order to establish a basis for the current development of a process guideline for the rehabilitation of patients with depressive disorders. METHOD: In order to identify evidence based treatment elements, first a comprehensive investigation of national and international guidelines was conducted. Thirteen selected guidelines were then assessed with regard to aspects of methodological quality and evidence-based treatment elements. In a further step, literature searches were conducted for residual treatment elements, which were identified on the basis of the Classification of Therapeutic Services (KTL) 2007. For the literature search, a hierarchical approach was chosen: At first, meta-analyses and systematic reviews were viewed. In case when there was still a lack of evidence for specific, potentially relevant treatment elements, the search was expanded to the level of primary studies. All selected reviews and primary studies then underwent a standardized assessment especially regarding methodological quality and evidence grades were allocated to treatments. RESULTS: Thereby, the following treatment elements with an adequate level of evidence were identified: Psychotherapeutic interventions, marital/couples/family therapy and counselling, inclusion of family members, psycho education and exercise, problem solving therapy, guided self-help, and behavioural activation treatments. On the basis of this complementary literature search, various other evident interventions could be identified within the following areas: relaxation techniques, improvement of social competence, occupational therapy, art therapies (music, movement/dance therapies), body-oriented therapies and massage therapy. CONCLUSION: In summary, using this hierarchical approach, it was possible to assign different levels of evidence to the various treatment elements for depression. Based on the results of this literature search, a next step in the development of a process guideline for the rehabilitative treatment of patients with depression will be the integration of experts in the field of rehabilitation. PMID- 20013573 TI - Daptomycin use in a neonate: serum level monitoring and outcome. AB - We present a case of successfully treated persistent bacteremia due to Gram positive bacteria with daptomycin in a preterm neonate. Daptomycin was given at higher doses (6 mg/kg/dose) and shorter intervals (every 12 hours) than those recommended for adults with no adverse events. Peak and trough serum concentrations of daptomycin were 27.3 and 11.6 MUg/mL [DOSAGE ERROR CORRECTED] at day 4 as well as 22.9 and 7.9 MUg/mL [DOSAGE ERROR CORRECTED]at day 11 after initiation of treatment, respectively. PMID- 20013574 TI - Obstetric outcomes in normal weight and obese women in relation to gestational weight gain: comparison between Institute of Medicine guidelines and Cedergren criteria. AB - We compared obstetric outcomes based on gestational weight gain in normal-weight and obese women using traditional Institute of Medicine (IOM) guidelines and newly recommended Cedergren criteria. Using the New Jersey Pregnancy Risk Assessment Monitoring System (PRAMS) database and electronic birth records, perinatal outcomes were analyzed to estimate the independent effects of prepregnancy body mass index (BMI) and gestational weight gain by IOM versus Cedergren criteria. Of 9125 subjects in PRAMS database from 2002 to 2006, 53.7% had normal BMI, 12.3% were overweight, 18.2% were obese, and the rest were underweight. Among normal-weight mothers, when compared with the IOM guidelines, macrosomia (6.45% versus 4.27%) and cesarean delivery rates (30.42% versus 29.83%) were lower using Cedergren criteria but the rates of preterm delivery (5.06% versus 9.44%), low birth weight (0.38% versus 2.42%), and neonatal intensive care unit (NICU) admissions (7.02% versus 10.86%) were higher with the Cedergren criteria. Similarly, among obese patients, when compared with IOM guidelines, macrosomia (10.79% versus 5.47%) and cesarean delivery rates (43.95% versus 40.71%) were lower using Cedergren criteria but the rates of preterm delivery (6.83% versus 8.32%), low birth weight (0.87% versus 1.88%), and NICU admissions (8.92% versus 13.78%) were higher with the Cedergren criteria. Based on our results, ideal gestational weight gain is presumably somewhere between the IOM and Cedergren's guidelines. PMID- 20013575 TI - Short-term outcomes of patent ductus arteriosus ligation in preterm neonates: reason for concern? AB - We evaluated outcomes of preterm infants following surgical ligation of patent ductus arteriosus (PDA). We performed a retrospective chart review. Our cohort (N = 82) had a median (range) gestational age of 25.5 (23 to 28) weeks and birth weight of 765 (484 to 1150) g. Ligation was the initial intervention in the primary group (N = 28) and was performed after failed medical therapy in the secondary group (N = 54). At 48 hours following ligation, an increase in fraction of inspired oxygen (FiO(2)) and ventilatory and pressor support were observed in 49 (60%), 35 (43%), and 15 (18%) infants, respectively. The median (range) duration to return to preoperative FiO(2) was 3 (1 to 30) days and extubation was 16 (1 to 98) days. Rates of bronchopulmonary dysplasia (BPD), severe intraventricular hemorrhage, necrotizing enterocolitis, and retinopathy were 42.7%, 32.9%, 19.6%, and 11%, respectively. Outcomes did not differ between primary and secondary ligation groups, nor between early (4 weeks old, N = 30). A lower gestation was significantly associated with postoperative deterioration and BPD. Preterm infants undergoing PDA ligation often experience a postoperative deterioration and have an extremely high incidence of comorbidities. Whether the ductus itself or the "therapeutic" ligation is causally linked to these adverse outcomes remains to be determined. PMID- 20013576 TI - Blood carbon dioxide levels and adverse outcome in neonatal hypoxic-ischemic encephalopathy. AB - We investigated pCO(2) patterns and the relationship between pCO(2) levels and neurodevelopmental outcome in term infants with hypoxic-ischemic encephalopathy. Blood gases during the first 72 hours of life were collected from 52 infants with hypoxic-ischemic encephalopathy. Moderate hypocapnia (pCO(2) <3.3 kPa), severe hypocapnia (pCO(2) <2.6 kPa), and hypercapnia (pCO(2) >6.6 kPa) were correlated to neurodevelopmental outcome at 24 months. Normocapnia was documented in 416/551 (75.5%) of samples and was present during the entire 72 hours in only 6 out of 52 infants. Mean (standard deviation) pCO(2) values did not differ between infants with normal and abnormal outcomes: 5.43 (2.4) and 5.41 (2.03), respectively. There was no significant association between moderate hypocapnia, severe hypocapnia, or hypercapnia and adverse outcome (odds ratio [OR] = 1.84, 95% confidence interval [CI] = 0.49 to 6.89; OR = 3.16, CI = 0.14 to 28.45; and OR = 1.07, CI = 0.24 to 5.45, respectively). In conclusion, only one in nine newborns had normocapnia throughout the first 72 hours. Severe hypocapnia was rare and occurred only in ventilated babies. Hypercapnia and hypocapnia in infants with hypoxic-ischemic encephalopathy during the first 72 hours of life were not associated with adverse outcome. PMID- 20013577 TI - Baseline illness severity does not alter accuracy of neonatal sepsis screen. AB - We investigated the relationship between illness severity and accuracy of neonatal sepsis screen. Consecutive neonates with clinically suspected early onset sepsis (EOS) were enrolled and blood culture and sepsis screen [C-reactive protein, absolute neutrophil count, immature to total ratio (ITR) and microerythrocyte sedimentation rate] were performed. Exclusion criteria were prior antibiotic exposure, nonavailable reports, and contaminated cultures. Score for Neonatal Acute Physiology Perinatal Extension (SNAPPE-II) was used to categorize neonates into "mild to moderate" (score 40) illness. Sepsis was defined as positive blood culture, and positive screen as >or=2 parameters positive. Of 125 subjects, 86 had mild to moderate and 39, severe illness. Twenty-eight (22%) subjects had sepsis. Sensitivity, specificity, negative predictive value, positive predictive value, likelihood ratio of positive test, and likelihood ratio of negative test of the sepsis screen and screen parameters were similar between the two groups. The sensitivity of the screen was 37.5% and 25% for mild to moderate illness and severe illness, respectively. Only ITR values correlated with SNAPPE-II scores in patients with "sepsis" (rho 0.4; P = 0.036). The severity of underlying illness does not alter the performance of the sepsis screen in diagnosing culture-positive EOS. PMID- 20013578 TI - Treatment of patent ductus arteriosus: indomethacin or ibuprofen? AB - We compared ibuprofen and indomethacin for the treatment of patent ductus arteriosus (PDA) in preterm infants. A retrospective comparative study was conducted at a pediatric tertiary center in preterm infants diagnosed with PDA. Infants born from January 2000 to June 2003 were treated with indomethacin, whereas infants born from July 2003 to November 2005 were treated with ibuprofen. The two treatment groups were compared. Demographic data and clinical, laboratory, and outcome data were collected from the medical files. Seventy-three infants were included in the ibuprofen group and 46 in the indomethacin group. No significant difference in efficacy was found between indomethacin and ibuprofen. Compared with ibuprofen, indomethacin treatment was associated with significantly higher mean creatinine levels and a higher percent of infants with creatinine >1.2 mg/dL, hyponatremia <120 mmol/L, and platelet level <100,000 platelets/mL(3). There were no significant differences in bilirubin levels, incidence and grade of intraventricular hemorrhage, necrotizing enterocolitis, retinopathy of prematurity, rate of surgical duct ligation, sepsis, length of hospital stay, or mortality. Indomethacin and ibuprofen are equally effective for PDA closure in premature infants. Treatment with ibuprofen is safer, decreasing the risk of renal failure, thrombocytopenia, and hyponatremia. PMID- 20013579 TI - Early versus delayed minimal enteral feeding and risk for necrotizing enterocolitis in preterm growth-restricted infants with abnormal antenatal Doppler results. AB - We studied the effect of early (< or = 5 days) versus delayed (> or = 6 days) initiation of minimal enteral feeding (MEF) on the incidence of necrotizing enterocolitis (NEC) and feeding intolerance in preterm infants with intrauterine growth restriction (IUGR) and abnormal antenatal Doppler results. We performed a randomized, nonblinded pilot trial of infants receiving early or delayed MEF in addition to parenteral feeding within 48 hours of life. Demographic data, maternal preeclampsia, antenatal steroid exposure, Doppler studies, as well as cases of NEC and feeding intolerance were all recorded. Of the 84 infants enrolled, 81 completed the study: 40 received early (median age: 2 days, range: 1 to 5 days) and 41 delayed (median age: 7 days, range: 6 to 14 days) MEF. The incidence of NEC and feeding intolerance was not significantly different between groups (p = 0.353 and p = 0.533, respectively). Birth weight was an independent risk factor for NEC in both groups. Early MEF of preterm infants with IUGR and abnormal antenatal Doppler results may not have a significant effect on the incidence of NEC or feeding intolerance. Furthermore, birth weight seems to be an independent risk factor for the development of NEC, irrespectively of the timing of MEF introduction. PMID- 20013580 TI - The risk for preterm labor in women receiving 17 alpha-hydroxyprogesterone caproate prophylaxis for preterm birth prevention. AB - We sought to identify maternal or clinical characteristics of women likely to develop preterm labor (PTL) at <34 weeks' gestation while receiving 17 alpha hydroxyprogesterone caproate (17P) prophylaxis. Current singleton gestations with prior preterm delivery enrolled for outpatient 17P administration at <27 weeks' gestation were identified ( N = 1177). Maternal and clinical characteristics were compared between women hospitalized and diagnosed with PTL at <34 weeks' gestation (PTL group, N = 270) and those without PTL (No PTL group, N = 660). PTL at <34 weeks' gestation occurred in 270/1177 (22.9%) of patients receiving 17P prophylaxis (mean gestational age at diagnosis was 28.3 +/- 4.0 weeks). Recurrent preterm delivery occurred in 73.3% of women with PTL at <34 weeks. Maternal age, marital status, race, tobacco use, cerclage, gestational age at start of 17P, and Medicaid status were similar between the groups. Women developing PTL at <34 weeks were more likely to have >1 prior preterm delivery than those without PTL (35.2% versus 25.9%, P = 0.006, odds ratio [95% confidence interval] 1.5 [1.1, 2.1]). Women receiving 17P prophylaxis remain at increased risk for PTL and preterm birth. Patient education and surveillance for PTL symptoms may be warranted in women with a history of more than one prior preterm delivery. PMID- 20013581 TI - Excessive weight gain among obese women and pregnancy outcomes. AB - We evaluated pregnancy outcomes in obese women with excessive weight gain during pregnancy. A retrospective study was performed on all obese women. Outcomes included rates of preeclampsia (PEC), gestational diabetes, cesarean delivery (CD), preterm delivery, low birth weight, very low birth weight, macrosomia, 5 minute Apgar score of <7, and neonatal intensive care unit (NICU) admission and were stratified by body mass index (BMI) groups class I (BMI 30 to 35.9 kg/m(2)), class II (36 to 39.9 kg/m(2)), and class III (>or=40 kg/m(2)). Gestational weight change was abstracted from the mother's medical chart and was divided into four categories: weight loss, weight gain of up to 14.9 pounds, weight gain of 15 to 24.9 pounds, and weight gain of more than 25 pounds. A total 20,823 obese women were eligible for the study. Univariate analysis revealed higher rates of preeclampsia, gestational diabetes, Cesarean deliveries, preterm deliveries, low birth weight, macrosomia, and NICU admission in class II and class III obese women when compared with class I women. When different patterns of weight gain were used as in the logistic regression model, rates of PEC and CD were increased. Excessive weight gain among obese women is associated with adverse outcomes with a higher risk as BMI increases. PMID- 20013582 TI - White's classification of diabetes in pregnancy in the 21st century: is it still valid? AB - White's classification system (WCS) was created 60 years ago to identify diabetic (DM) pregnancies at increased risk for perinatal morbidity and mortality. Our objective was to assess the association between WCS and adverse pregnancy outcome (APO) in contemporary DM pregnancies. We studied diabetic women with singleton pregnancies who delivered at >20 weeks at a single institution over a 1-year period (2007 to 2008). Perinatal outcomes were compared between WCS groups. APO was defined as any of the following: preterm birth <34 weeks, severe preeclampsia, shoulder dystocia, and neonatal respiratory disease. Presence of vascular disease was defined as presence of chronic hypertension, chronic renal insufficiency, retinopathy, coronary artery disease, or prior cerebrovascular event. One hundred ninety-six DM pregnancies met the criteria. No significant differences in APO existed between White's class groups among women with pregestational DM (32.7% class B versus 26.9% class C versus 57.1% class D to F; p = 0.46). Logistic regression revealed that vascular disease was associated with APO (odds ratio = 2.7, 95% confidence interval = 1.2 to 6.2). In our population, presence of vascular disease, rather than WCS, was a better predictor of APO in DM women. PMID- 20013584 TI - Intrauterine exposure to tobacco and risk of medically indicated and spontaneous preterm birth. AB - We investigated the association between prenatal smoking and the occurrence of medically indicated and spontaneous preterm delivery (<37 weeks). We performed a retrospective cohort study of singleton live births in the state of Missouri (n = 1,219,159) using maternally linked cohort data files covering the period 1989 to 2005. The main outcomes of interest were spontaneous and medically indicated preterm and very preterm birth. Logistic regression models were used to generate adjusted odds ratios and their 95% confidence intervals. There were 132,246 (10.8%) infants born preterm in the study population, of which 106,410 (80.5%) were classifiable as spontaneous preterm births and 25,836 (19.5%) were medically indicated preterm deliveries. We found elevated risks for both medically indicated and spontaneous preterm birth associated with maternal cigarette smoking during pregnancy. This heightened risk was particularly evident for medically indicated preterm birth (adjusted odds ratio [95% confidence interval] = 1.48 [1.41 to 1.55]). Women who smoke during pregnancy are at increased risk for preterm birth, and especially for medically indicated preterm delivery. PMID- 20013583 TI - Unconjugated hyperbilirubinemia and early childhood caries in a diverse group of neonates. AB - We evaluated if the development of early childhood caries is associated with the severity of unconjugated hyperbilirubinemia during the first 2 weeks after birth. We performed a retrospective case-control study of children less than 6 years of age seen for comprehensive dental examination by pediatric dentists years following a hospital stay in the neonatal intensive care unit. Exclusion criteria included genetic disorders, cleft palate, direct hyperbilirubinemia, and missing information on jaundice. Children with early childhood caries were compared with those without dental caries for a panel of perinatal and neonatal clinical variables. Seventy-six children met study criteria. Of 76 children, 42 children had early childhood caries, while 34 children had healthy primary dentitions. Among clinical variables, only race and peak total serum bilirubin concentration differed significantly between the two groups on bivariate analysis. On logistic regression, peak total serum bilirubin concentration was significantly associated with early childhood caries (adjusted odds ratio 1.17, 95% confidence interval 1.04 to 1.32). Neonatal unconjugated hyperbilirubinemia may be associated with early childhood caries in children. PMID- 20013585 TI - Maternal blood pressure adaptation in the first trimester of pregnancy. AB - Normal changes in blood pressure during pregnancy are well documented in the second and third trimesters. Little is known about first-trimester changes, particularly compared with preconceptional values. This knowledge might allow for early prediction of conditions such as preeclampsia or intrauterine growth restriction. Prior studies utilized a laboratory setting. We conducted this retrospective study to compare blood pressure readings in early pregnancy with preconceptional values in the clinical setting. The records of 44 healthy normotensive nonsmoking women with a body mass index <30 and an uncomplicated appropriately grown singleton term delivery were reviewed. Preconceptional blood pressures values were compared with values at five periods (weeks 1 to 8, 9 to 16, 17 to 23, 24 to 32, 33 to 40). There was no difference in blood pressure parameters when comparing preconceptional values with period 1. The mean and systolic arterial pressures decreased significantly in periods 2 and 3. We were unable to demonstrate significant change in any blood pressure parameter in the first 8 weeks of pregnancy. This work should be repeated in patients who subsequently develop preeclampsia, spontaneous abortion, or intrauterine growth restriction to determine if early changes might be evident in early pregnancy to identify patients destined to develop pregnancy complications. PMID- 20013586 TI - Clinical application of the anterolateral thigh flap for soft tissue reconstruction. AB - The purpose of this article is to describe the author's experience using the anterolateral thigh (ALT) flap for the reconstruction of a variety of soft tissue defects. The flap utility and donor site morbidity were evaluated in 126 cases from March 1985 to August 2007. The ALT flaps were harvested as either free fasciocutaneous, free adipofascial, fasciocutaneous island, or reversed fasciocutaneous island flaps to repair facial, neck, breast, trunk, and extremity defects. In 40 cases (32%), the skin vessels were found to be septocutaneous perforators, and in 86 cases (68%), they were found as musculocutaneous perforators. Of the 126 flaps, 121 survived completely, providing a success rate of 96.0%. There were four cases undergoing multidetector-row computed tomographic angiography (CTA) for preoperative perforator mapping, and all perforators were confirmed intraoperatively. In conclusion, the ALT flap is a versatile and reliable flap that could well be a priority option for soft tissue reconstruction. CTA can provide more valuable and accurate anatomic information about the pedicle and perforators, making it safer and faster to harvest a targeted ALT perforator flap with less donor site morbidity. PMID- 20013587 TI - A novel approach for preventing the development of persistent vasospasms after microsurgery for the extremities: intermittent topical lidocaine application. AB - Vasospasm is often encountered after harvesting the recipient artery in tissue transfer surgery, particularly in the extremities. Further, after anastomosis, thrombosis is a major complication arising due to vasoconstriction. Therefore, we decided to apply lidocaine topically on the recipient artery to prevent postoperative vasospasm. We had applied lidocaine topically on the recipient artery in seven patients with persistent vasospasm before the completion of the surgery. After surgery, 0.2 mL lidocaine (4%) was directly applied on the vascularized region every 15 minutes for the first 3 hours, every 30 minutes for the next 3 hours, and every 1 hour for the next 18 hours. Although four patients experienced a disturbance in the blood flow immediately after the surgery, they showed improvements after lidocaine application. In three of these four patients, vasospasm was also suspected to occur at 15 minutes after the surgery; however, topical application of lidocaine was found to be effective in these patients. Thrombosis was not observed in any case. We believe that if the site of lidocaine application is appropriate, even a small dose of low-concentration lidocaine (4%) can produce an adequate effect. Moreover, the intervals between lidocaine applications should be carefully considered. PMID- 20013588 TI - Homeochronous usage of structural bone allografts with vascularized fibular autografts for biological repair of massive bone defects in the lower extremities after bone tumor excision. AB - We report the clinical outcomes of homeochronous usage of massive bone allografts with vascularized fibular autografts for the biological repair of lower-extremity bone defects secondary to bone tumor excision. Large bone defects in the lower extremities of 17 patients (10 men and seven women; age range, 6 to 34 years) who underwent bone tumor excision were repaired using massive bone allografts along with vascularized fibular autografts. After 6 to 48 months (mean, 20.2 months) of follow-up, the wounds at the donor and recipient sites healed in one stage, the monitoring flaps remained viable, the immune response to the massive bone allografts was minimal, and there were no complications in the donor limbs. Of the 17 patients, 13 and 3 cases showed radiographic union in 6 months and 8 months after surgery, respectively; one case of malignant synovioma showed recurrence, and therefore, leg amputation was performed 2.5 months after surgery. None of the massive bone allografts were absorbed or fractured at the last follow up. The Mankin evaluation rating was excellent in eight cases, good in five cases, fair in two cases, and poor in two cases; the total rate of excellent and good cases was 76.47%. Homeochronous usage of massive bone allografts with vascularized fibular autografts is a satisfactory method for the biological repair of massive bone defects in the lower limbs after bone tumor excision. PMID- 20013589 TI - Specific volume of female subcutaneous abdominal tissue as a reference in autologous breast reconstruction. AB - Use of the deep inferior epigastric perforator (DIEP) flap is gaining popularity as the method of choice for breast reconstruction and replacement of prosthetic breast implants. Ideally, the volume of the prosthesis is replaced by the same volume of autologous tissue to restore shape and symmetry. Still, intraoperative tissue volumetry is not practical under sterile circumstances. The first step toward a clinical intraoperative measure for volume matching would be to acquire knowledge of the specific density of female subcutaneous abdominal fat tissue. The weight and volume of abdominoplasty specimens from 32 women were measured. From these data, the specific volume of DIEP flap tissue was calculated. The mean specific density was 1.12 mL/g (range, 1.02 to 1.32 mL/g; standard deviation 0.04). An approximate volume-to-weight conversion multiplier of 1.1 may serve as an intraoperative tool to estimate the volume of a DIEP flap from its weight. PMID- 20013590 TI - Measuring the pressure in the superficial inferior epigastric vein to monitor for venous congestion in deep inferior epigastric artery perforator breast reconstructions: a pilot study. AB - During deep inferior epigastric artery perforator (DIEP) flap dissection, we noted that in many cases the superficial vein on the ipsilateral side of the flap was engorged and tense, and in others, it was empty. This led us to believe that the pressure is increased as the result of preferential outflow through the superficial vein in some cases, which could result in venous congestion of the flap if this vessel was not anastomosed. To test this hypothesis, we measured the venous pressure in the superficial venous system before and after flap dissection. The pressure in the superficial inferior epigastic vein of a DIEP flap was measured in 26 consecutive flaps to investigate the correlation between the pressure and venous congestion of the flap. The first measurement was performed at the beginning of the dissection, and the second measurement was taken after the flap had been completely raised on a single perforator. The mean increase in pressure after flap dissection was 10.6 mm Hg (mu = 10.6; range -1 to 31; O +/- 7.0 mm Hg). Clinical signs of venous congestion were observed in one case. In this case, the increase in venous pressure was with 31 mm Hg, also the highest. Although the results of this report are preliminary, they indicate that the pressure in the superficial vein of DIEP flaps might be of predictive value for venous congestion. PMID- 20013591 TI - Allografting combined with systemic FK506 produces greater functional recovery than conduit implantation in a rat model of sciatic nerve injury. AB - Implantation of allografts or nerve conduits has been used to promote regeneration following peripheral nerve injuries involving substantial axon loss. Both methods provide promising alternatives to autologous grafting and avoid donor site morbidity. We compared the relative efficacies of allografting versus conduit implantation in a rat model of sciatic nerve regeneration. Two rat strains (Lewis and Dark Agouti; n = 30) were employed. Unoperated animals served as controls (group I). Animals in groups II and III underwent left sciatic nerve resection over a distance of 15 mm; group II animals received implants of collagen type I conduits; and group III animals received allografts from the other rat strain and systemic low-dose (0.1 mg/kg/d) administration of FK506. Walking tracks were recorded after 4, 8, 12, and 16 weeks; nerve sections were stained for myelin basic protein after 16 weeks. Functional tests revealed significantly better recovery in group III animals compared with group II even though there was no significant difference in the extent of remyelination. Neither group achieved the functional or histomorphometric values of control animals. Improved functional recovery following allografting plus systemic FK506, in comparison with conduit implantation, underlines the importance of systemic administration of neurotrophic molecules for nerve regeneration. PMID- 20013592 TI - Further validation for use of the retrograde limb of the internal mammary vein in deep inferior epigastric perforator flap breast reconstruction using laser assisted indocyanine green angiography. AB - We have previously described the use of the retrograde limb of the internal mammary vein (IMV) as an additional venous outflow tract in deep inferior epigastric perforator (DIEP) flap reconstruction. In the current study, we use the Novadaq SPY ((R)) system, a novel intraoperative angiographic method, to further validate the use of the retrograde limb of the IMV. The Novadaq SPY ((R)) system laser source was used with indocyanine green angiography to evaluate the arterial, anterograde venous, and retrograde venous anastomoses of 15 patients undergoing DIEP flap breast reconstruction. The number of perforators used, patient age, exposure to radiation, coupler size, and incidence of intraoperative congestion were recorded. All flaps survived, and there were no cases of intraoperative congestion. The average time required to perform the additional retrograde anastomosis was 12 minutes. Flow of indocyanine green not only revealed patency of our anastomoses but it confirmed unobstructed flow through the retrograde limb of the IMV. Our study further validates that the retrograde limb of the IMV does in fact achieve flow away from the DIEP flap and can therefore be used as an additional or alternative outflow tract in DIEP flap breast reconstruction surgery. PMID- 20013593 TI - Extracorporeal shock wave treatment in ischemic tissues: what is the appropriate number of shock wave impulses? AB - The dose-dependent effect of extracorporeal shock wave technology (ESWT) was evaluated using a murine skin flap model. Thirty-six Sprague-Dawley rats were divided into six groups (ESWT groups 1 through 5 and a control group). After surgery, shock wave impulses doses were administered: 200 (group 1), 500 (group 2), 1500 (group 3), 2500 (group 4), 5000 (group 5), and 0 (control group 6). Flap viability was evaluated on day 7. Overall, significantly smaller percentages of necrotic zones were observed in groups 2, 3, and 4 compared with groups 1, 5, and the control group ( P < 0.05). ESWT treatment with 200 impulses was found to be ineffective. ESWT treatment of 5000 impulses resulted in a significant increase in the percentage of necrosis compared with other ESWT groups ( P < 0.05). However, ESWT treatments between 500 and 2500 impulses at 0.11 mJ/mm (2) enhanced epigastric skin flap survival significantly. PMID- 20013594 TI - Aberrant perfusion of the serratus anterior muscle flap: report of two cases and a review of the literature. AB - Free serratus anterior muscle flap, classified as Mathes-Nahai type III, is a versatile and frequently preferred choice for soft tissue coverage of small to moderate-sized defects owing to its reliable circulation pattern; however, some anatomic variations in the flap vascularity can result in partial flap loss. Here we present two cases with free serratus anterior muscle transfer where the distalmost portion of the flap was not perfused by the thoracodorsal pedicle. The most likely explanation seems to be the large flap size and the lack of distal interconnections between lateral thoracic artery and thoracodorsal artery branches. Our clinical experience suggests that anatomic variations of serratus anterior muscle circulation might end up with distal perfusion loss if a large flap is to be harvested. PMID- 20013595 TI - Modified approach to construct a vascularized coral bone in rabbit using an arteriovenous loop. AB - The most important factor for the survival of thick three-dimensional tissues is the degree of vascularization. In this study, a modified arteriovenous loop (AVL) model was developed to prefabricate an axial vascularized tissue-engineered coral bone. In group A (n = 28), an arteriovenous fistula between rabbit femoral artery and vein was anastomosed to form an AVL. The AVL was placed in a coral block (6 x 8 x 10 mm (3)) as a vascular carrier. The complex was wrapped with polytetrafluoroethylene membrane and implanted subcutaneously. In group B (n = 20), there was no vascular carrier, and the same dimensional coral was directly implanted beneath inguinal skin. After 2, 4, 6, and 8 weeks, the rabbits were perfused with heparinized saline (for scanning electron microscopy), India ink (for histological examination), and ethylene perchloride (for vascular casts) via the abdominal aorta. In group A, histology showed that newly formed vasculature extended over the surfaces and invaded the entire coral blocks. The vascular density was significantly superior to that in group B. Vascular casts showed that new blood vessels robustly sprouted from the AVL. Scanning electron microscopy demonstrated that there were minute sprouting cavities in the vascular endangium. In this model, an axial vascularized coral bone could be effectively constructed. PMID- 20013596 TI - Comparison of primary and secondary lower-extremity lymphedema treated with supermicrosurgical lymphaticovenous anastomosis and lymphaticovenous implantation. AB - Although some authors previously stated that microlymphatic surgery does not have application to primary lymphedema, opposite views are reported based on the observations that the lymphatics were not hypoplastic in majority of these patients and microlymphatic surgery yielded significant improvement. The aim of this study was to compare the intraoperative findings and outcomes of primary and secondary lower-extremity lymphedema cases treated with lymphaticovenous shunts. Between December 2006 and April 2009, microlymphatic surgery was performed in 80 lower extremities with primary and 21 with secondary lymphedema. These two groups of extremities are compared according to the morphology of the lymphatic vessels and possibility of precise anastomoses, their response to the treatment, and final outcomes based on volumetric measurements during the follow-up period. The morphology of the lymphatics in secondary lymphedema was more consistent, and at least one collector larger than 0.3 mm was available for anastomosis in 20 of 21 extremities. In the primary lymphedema group, the lymphatics were smaller than 0.3 mm in 13 of 80 extremities. It was, therefore, possible to perform supermicrosurgical lymphaticovenous anastomosis in 84% of extremities with primary lymphedema and 95% of extremities with secondary lymphedema. Reduction of the edema occurred earlier in the secondary lymphedema group, but the mean reduction in the edema volume was comparable between the two groups. Microlymphatic surgery, although more effective and offered as the treatment of choice for secondary lymphedema, would also be a valuable and relevant treatment of primary lymphedema. PMID- 20013597 TI - [Cancer information service--not just for patients]. PMID- 20013598 TI - [AB0-incompatible kidney transplantation--case 13/2009]. AB - PATIENT HISTORY AND CLINICAL FINDINGS: A 49-year old patient with chronic kidney disease was referred for evaluation of living donor kidney transplantation from his spouse in a blood group incompatible setting. CLINICAL INVESTIGATIONS: Patient blood group was 0, donor blood group was A, subtype A. recipient isoagglutinin titer against donor erythrocytes was 1:64. Crossmatch (CDC) and antibody screening were negative. There were no contraindications for living donor kidney transplantation from donor and recipient side. DIAGNOSIS, TREATMENT AND CLINICAL COURSE: For AB0-incompatible living donor kidney transplantation, recipient preconditioning using rituximab and selective immunoadsorption was initiated to obtain reduction of isoagglutinin titers < or = 1:4. Transplantation was performed without complications. With only moderate increases in titer, no immunoadsorption was required postoperatively. Monitoring of isoagglutinin titers was discontinued after day 14, since increasing titers do not result in organ loss any more (accomodation). 18 months after transplantation, renal function is excellent under corticosteroid-free maintenance immunosuppression. CONCLUSION: Recipient preconditioning nowadays allows successful blood group incompatible kidney transplantation in most cases, increasing the number of patients eligible for living donor kidney transplantation by up to 20%. No intensified maintenance immunosuppression is required and renal allograft function after AB0-incompatible transplantation is comparable to blood group compatible living donor kidney transplantation. PMID- 20013600 TI - Intestinal ischemia-reperfusion injury causes activation of bone marrow-derived progenitor cells. AB - INTRODUCTION: Bone marrow-derived circulating granulocyte and macrophage progenitor cells can contribute to the regeneration of ischemic tissue. Mobilization after heart or brain ischemia is well established, but it is unclear if this occurs after intestinal ischemia-reperfusion injury. Our aim was to evaluate bone marrow granulocyte-macrophage proliferation and the possible beneficial effect of recombinant human granulocyte-colony stimulating factor (rhG CSF) in a model of intestinal ischemia-reperfusion. MATERIAL AND METHODS: After animal committee approval, anesthetized adult rats were divided into groups (n=4 per group) as follows: (i) control [C], (ii) 60 min intestinal ischemia [I], (iii) 60 min intestinal ischemia+360 min reperfusion [IR], (iv) 420 min sham operation [SH]. At sacrifice, bone marrow was removed, erythrocytes lysed and 1 50 000 nucleated cells plated in triplicate in 35 mm Petri dishes containing methylcellulose (MethoCult). After 11 days, granulocyte-macrophage colony-forming units (CFU-GM) were counted. In addition, to determine whether rhG-CSF injection stimulates progenitor cell activation, two further groups were studied: (v) 60 min intestinal ischemia+360 min reperfusion with injection of 50 microg/kg rhG CSF at reperfusion [IR-G]; (vi) 420 min sham with rhG-CSF injected at 60 min [SH G]. Data are expressed as median, range and IQR and compared using one-way ANOVA with Tukey's post-hoc test. RESULTS: Neither sham operation nor ischemia alone influenced the activation of bone marrow. However, IR caused a significant increase in bone marrow activation compared to control animals (p<0.01), ischemic animals (p<0.01) and sham operated animals (p<0.05). Administered at a dose of 50 microg/kg, which is commonly used in animal studies, rhG-CSF had no effect on bone marrow activation, and did not augment the effects of ischemia-reperfusion. At a higher dose (100 microg/kg), however, rhG-CSF resulted in the mortality of IR animals. CONCLUSIONS: Intestinal ischemia-reperfusion injury causes proliferation of bone marrow granulocyte-macrophage progenitors which contribute to long-term repair. This phenomenon is not augmented by the administration of exogenous rhG-CSF. PMID- 20013601 TI - Use of the TIP principle for the repair of non-glanular recurrent post hypospadias urethrocutaneous mega fistula. AB - INTRODUCTION: The most significant complication after hypospadias repair is urethrocutaneous fistula. Repair is even more difficult if the fistula is large. Use of the tubularized incised plate (TIP) procedure for hypospadias repair has greatly increased. However, use of the TIP procedure for the repair of recurrent mega fistula has not been previously described. The aim of this study is to present the results of a modified TIP procedure for the repair of recurrent mega fistula occurring after hypospadias repair. METHODS: All cases of recurrent penile mega fistula after hypospadias repair presenting to our institution between 2002 and 2008 were included in our study. Cases with coronal or glanular fistulae were excluded. Diameters of these large fistulae were > or =0.5 cm. Repair was done a minimum of 6 months after the last repair. After complete dissection of the fistula, the excision was extended a further 2 mm all around the edge in a circumferential manner. A dorsal slit in the penile urethra was made based on the TIP procedure; the urethroplasty was completed using interrupted sutures. Second layer coverage was done and the skin was closed. RESULTS: A total of 11 boys (median age at surgery: 8 years) who developed mega fistula after hypospadias repair were included in the study. The patients had undergone previous attempts at repair, with the number of previous attempts ranging between 1 and 9 times. The last repair was done 6-48 months before surgery (median: 16 months). Fistulae diameters were between 5 and 13 mm (median: 9 mm). All patients underwent the same procedure, with a follow-up period of between 6 and 72 months (median: 33 months). In 7 cases 2 layers were used to cover the urethroplasty, while in 4 cases used only one layer. There were no intraoperative complications. Two cases suffered superficial infection postoperatively, one of whom developed a small fistula (1/11). CONCLUSIONS: The advantages of a procedure based on the TIP principle for the treatment of recurrent penile mega fistula are numerous. The procedure is easy to perform and can be successfully used to treat recurrent urethrocutaneous fistula in carefully selected cases. Our recurrence rate of 9% is acceptable. Use of a modified TIP procedure for the repair of mega fistula or partial penile disruption is feasible. More cases are needed to support our initial findings of this new use of the TIP procedure in hypospadias surgery. PMID- 20013602 TI - Lifestyle modification program decreases pregnancy weight gain in obese women. AB - We assessed the impact of a lifestyle modification program (LMP) on weight gain in pregnancy and evaluated its effect on adverse pregnancy outcomes. A prospective matched controlled study design was employed. Inclusion criteria consisted of pregnant women with singleton gestations and body mass index (BMI) > or = 30 without underlying medical conditions. In the study group, patients were prospectively enrolled in the first trimester. Through the LMP, women participated in a comprehensive program on nutrition, exercise, and weight control in pregnancy. The control group consisted of matched patients by starting BMI, parity, and socioeconomic status during the same study period cared for at the same institution. The primary outcome evaluated was weight gain in pregnancy. Adverse pregnancy outcomes including preeclampsia, gestational diabetes, gestational hypertension, and various postpartum complications were evaluated between the two groups. Statistical analysis was performed using nonparametric methods, with p < 0.5 considered significant. Twenty-eight patients were recruited for this study and underwent the intervention. Patient compliance with the LMP was 75%, two patients were lost to follow-up, one had a preterm delivery, and four had incomplete data due to various reasons. Twenty-one patients completed their pregnancy and were available for analysis. Twenty matched control patients were identified for analysis. Study participants gained significantly less weight in their pregnancies when compared with controls (mean weight gain 17 versus 34 pounds, respectively; p = 0.008). Secondary outcomes of preeclampsia, gestational diabetes, cesarean section, as well as infant birth weight did not significantly differ between the groups. This study suggests an effective method of reducing prenatal weight gain in the obese population. This type of comprehensive intervention could be an important, cost-effective risk-reduction strategy. PMID- 20013603 TI - Gum chewing slightly enhances early recovery from postoperative ileus after cesarean section: results of a prospective, randomized, controlled trial. AB - Postoperative ileus is one of the common problems after abdominal surgeries. It contributes to delayed recovery and prolongs hospital stay. Sham feeding, such as gum chewing, may accelerate return of bowel function and reduce morbidity and length of hospital stay. This study aimed to determine whether gum chewing in the immediate postoperative period facilitates a return to bowel function in cesarean delivery patients. Three hundred eighty-eight patients who underwent cesarean delivery were randomly assigned to a gum-chewing group (group G, N = 193) or a control group (group C, N = 195). Demographic data, duration of surgery, type of anesthesia, and time of discharge from hospital were recorded. Patients in the gum-chewing group chewed gum three times per day as soon as returning from the operating theater to the ward until the time they defecated or were discharged. Patients were asked to chew gum at least half an hour each time. The T test and Pearson chi-square test was used for statistical analysis. Groups were comparable in age, weight, height, weeks of gestation, duration of surgery, and type of anesthesia. Bowel sounds were 5 hours earlier in the gum-chewing group (mean 18.2 hours) than in the control group (mean 23.2 hours). Passing flatus was 5.3 hours earlier in group G (mean 34.6 hours) than in group C (mean 39.9 hours). Patients having mild ileus symptoms were 9% less in group G (mean 12%) than in group C (mean 21%). The difference between the two groups were all highly significant (p < 0.001). Gum chewing was easily tolerated without any complications. Gum chewing is an inexpensive, convenient, and physiological method in enhancing the recovery of bowel function. But this may not facilitate early hospital discharge, lactation, or defecation. PMID- 20013604 TI - The influence of phototherapy on serum cytokine concentrations in newborn infants. AB - We sought to determine cytokine response in term and late preterm newborn infants on phototherapy. Twenty newborn infants with gestational age > or = 35 weeks and birth weight > or = 2000 g in the first week of life had serum interleukin (IL) 6, IL-8, IL-10, IL-1beta, and tumor necrosis factor (TNF)-alpha measured immediately prior to and after 24 hours on phototherapy. Exclusion criteria were newborns with severe congenital malformations, congenital infections, birth asphyxia, sepsis, hemolytic anemia that required blood transfusion, maternal infant Rh incompatibility and those who required exchange transfusion or intravenous immunoglobulin treatment for hyperbilirubinemia. Median IL-6 concentrations significantly decreased after 24 hours on phototherapy (18.3 pg/mL and 7.85 pg/mL, respectively, p = 0.005). IL-6 concentrations decreased in 17 out of the 20 newborns. There were no statistical differences in IL-8, IL-10, IL 1beta, and TNF-alpha concentrations before and after 24 hours on phototherapy. There was a statistically significant correlation between IL-6 decline and irradiance (r = 0.57, p = 0.009). The finding that serum IL-6 decreases in newborn under phototherapy suggests that phototherapy possibly has an anti inflammatory effect, although the clinical implications of this study deserve further studies. PMID- 20013605 TI - Safety and effectiveness of indomethacin versus ibuprofen for treatment of patent ductus arteriosus. AB - We compared the rates of medical closure of patent ductus arteriosus (PDA) and complications (renal dysfunction, necrotizing enterocolitis, spontaneous intestinal perforation, and intraventricular hemorrhage) between infants treated with indomethacin and infants treated with ibuprofen. We performed a retrospective comparative cohort study of infants treated with indomethacin or ibuprofen for symptomatic PDA at Duke University Medical Center between November 2005 and November 2007. We identified 65 infants who received indomethacin and 57 who received ibuprofen. The rate of survival without surgical ductal ligation was 62% (40/65) in the indomethacin group and 58% (33/57) in the ibuprofen group (p = 0.71). The rate of the composite of complications (death, necrotizing enterocolitis, or intestinal perforation) was 40% (26/65) in the indomethacin group and 32% (18/57) in the ibuprofen group (p = 0.35). There was no significant difference between groups in elevation of serum creatinine during treatment. In clinical practice, ibuprofen appears to be as effective as indomethacin for closure of patent ductus arteriosus with similar complication rates. The decision to use one agent over the other should be based on dose schedule preference and the currently published clinical trials until more safety and effectiveness data are available. PMID- 20013606 TI - N-terminal pro-B-type natriuretic peptide as a marker of bronchopulmonary dysplasia in premature infants. AB - We performed an observational pilot study of plasma concentrations of N-terminal pro-B-type natriuretic peptide (BNP) in premature infants with a diagnosis of bronchopulmonary dysplasia (BPD) at 4 weeks of age and after 1 month of conventional therapy. Thirty-four premature infants born before 34 weeks' gestational age without cardiac or infectious diseases were included. Serum NT pro-BNP was measured in all neonates at 4 weeks of age. In infants with the diagnosis of BPD (n = 11), measurements were repeated at 6 and 8 weeks of age under conventional treatment. Specific clinical characteristics were collected prospectively. Baseline NT-pro-BNP concentrations were high in healthy premature infants compared with previously reported healthy neonates, and significantly higher in those who developed BPD. There was a significant correlation between concentrations of NT-pro-BNP and severity of respiratory distress as assessed by several methods. The concentrations of NT-pro-BNP decreased significantly over time in BPD infants. Premature infants have high concentrations of NT-pro-BNP at 1 month of age. NT-pro-BNP concentrations are significantly higher in BPD infants and decline over time. NT-pro-BNP concentrations correlate with clinical severity of respiratory disease. PMID- 20013607 TI - [Examples for asbestos-related findings in HRCT - criteria for the assessment of causal relationships in surveillance programmes and medical expert opinion]. AB - The increasing use of high-resolution computed tomography in formerly asbestos exposed workers requires valid diagnostic criteria for the findings which have to be reported as suspicious for being asbestos-related in surveillance programmes and for the assessment of causal relationships between former asbestos exposure and findings in computed tomography. The present article gives examples for asbestos-related findings in HR-CT and discusses the specificity of parenchymal and pleural changes due to asbestos fibres. PMID- 20013608 TI - Diagnostic accuracy of rhTSH test with neck ultrasonography in differentiated thyroid cancer follow-up. AB - Serum thyroglobulin levels measurement after injection of recombinant human thyrotropin (rh-TSH) represents the most important advance in the follow-up of patients with differentiated thyroid cancer, obtaining TSH elevation without L thyroxine withdrawal, avoiding marked hypothyroidism symptoms. During a 4-yr period (2004-2008), 66 consecutive patients with DTC (59 papillary and 7 follicular carcinomas) were examined after rh-TSH Tg test and neck ultrasonography. In all patients basal Tg was <0.25 ng/ml. In twelve (18.5%) examined patients rh-TSH Tg was >0.25 ng/ml, and in seven (58.3%) of these was demonstrated persistent or recurrent disease. These data indicate that rhTSH Tg>0.25 ng/ml should be considered diagnostic for persistent or recurrent disease and suggests further exams (neck ultrasonography, whole body scan or cytology) to localize the disease. Furthermore, neck ultrasonography has demonstrated high accuracy in detecting lymph nodal metastases and should be always combined with rh-TSH test. PMID- 20013609 TI - Ultrasound-assisted microsurgery for Cushing's disease. AB - OBJECTIVE: Localization of microadenomas in Cushing's disease may be difficult as in up to 45% of patients sellar MRI fails to detect a pituitary tumor. Intraoperative transsphenoidal ultrasound may identify microadenomas as hyperechoic structures. We report on the first 18 consecutive cases with intraoperative use of a new device for direct contact high-frequency-ultrasound in patients with Cushing's disease. PATIENTS AND TECHNIQUE: 18 patients (14 female, 4 male, age 24-71 years) with typical endocrinological findings for Cushing's disease were included in the study. One macroadenoma and 13 microadenomas were suspected or identified preoperatively by MRI. In 4 cases, two of them with recurrent disease, sellar MRIs were negative. During transsphenoidal microsurgery an end fire ultrasound-probe (B-mode frequency range 7.5-13 Mhz, field of view 5 mm, penetration 20 mm) was introduced after opening of sellar floor. The pituitary gland was scanned in direct contact to the capsule. RESULTS: In 13 out of 17 cases (77%) with later on proven microadenomas high-frequency ultrasound identified the tumors as hyperechoic masses, including 3 of the 4 cases with negative preoperative MRI. In 2 cases ultrasound correctly localized the tumor at a site different from MRI finding (MRI false positive). In the macroadenoma, identification of the border between tumor and anterior pituitary gland was not possible. In all 18 patients postoperative early decline of serum cortisol to subnormal levels confirmed remission of hypercortisolism (100%). Other pituitary functions were unaltered in 17 cases (94%). CONCLUSIONS: Intraoperative scanning of the pituitary gland with high-frequency-ultrasound probes may identify intrapituitary anatomy and pathologies even in MRI-negative cases. This may prevent extensive exploration of the gland with the risk of subsequent hypopituitarism. PMID- 20013610 TI - New mutations in the RET protooncogene-L881V - associated with medullary thyroid carcinoma and -R770Q - in a patient with mixed medullar/follicular thyroid tumour. AB - Clinical studies are needed to classify rare and novel RET mutations associated with hereditary medullary thyroid carcinoma (MTC) into one of the clinical risk groups. Here we describe two new RET mutations/variants, R770Q and L881V, in patients with MTC and analyzed genotype-phenotype correlations associated with these RET mutations in the gene carriers. FAMILY 1: Calcitonin screening in a 42 year-old female patient with multinodular goiter showed elevated levels. RET mutation analysis revealed a new variant in exon 13 R770Q (CGA>CAA) in the patient. A thyroidectomy with central and lateral node dissection was done. Histology showed MTC in a mixed variance with follicular cancer of 2 cm diameter (T2N0M0). Postoperatively there was no increase of calcitonin after pentagastrin stimulation. The patient is biochemically cured concerning MTC and FTC after radioiodine therapy. In the sister of the index patient surprisingly another, previously not described amino-acid substitution Y791N (TAT><) in the RET protooncogene was found. In the parents the R770Q variant was detected in the mother, the Y791N mutation in the father. Another sister carries the R770Q variant. In all other gene carriers (aged 44-70 years), calcitonin levels were in the normal range, therefore, thyroidectomy had not yet been performed. FAMILY 2: In a 46-year-old female patient with nodular goiter thyroidectomy, central and left lateral lymph node dissection was done because of elevated calcitonin levels. Histology revealed a microcarcinoma with one lymph node metastasis (T1N1(1/8)Mx). RET analysis revealed a new mutation in exon 15 L881V (CTG>GTG). The L881V mutation was detected in five other family members. In the first generation stimulated calcitonin levels were in the normal range, therefore thyroidectomy had not yet been performed. In the sons of the index case thyroidectomy revealed CCH in the older one, no MTC in both. In a cousin thyroidectomy is intended because of elevated basal and stimulated calcitonin. CONCLUSION: Our clinical findings indicate that the L881V mutation may be associated with late-onset nonaggressive disease. If the germline RET R770Q variant has a causative role in the pathogenesis of the mixed medullar/follicular derived histology of the thyroid tumour in the index patient of family 1 has to be proven. The recommendations for prophylactic thyroidectomy in these mutations should be individualized depending on basal and stimulated calcitonin levels until more data are available. PMID- 20013611 TI - Effects of long-term exposure to iodine excess on the apoptosis of thyrocytes in Wistar rats. AB - OBJECTIVE: To explore the effects of long-term exposure to more than adequate and excessive iodine on the apoptosis of thyrocytes in Wistar rats. METHODS: Wistar rats, divided into four groups: Group 0 (control, treated with 4 MUg/day of iodine), Group 1 (1.5x control dose, i. e. 6 MUg/day of iodine), Group 2 (3x control dose, i. e. 12 MUg/day of iodine), and Group 3 (6x control dose, i. e. 24 MUg/day of iodine) had oral iodine intake for 1, 2, 4, and 8 months before being sacrificed for study. After the initial treatment course of 8 months, some rats in each group continued to have an iodine intake at 4 MUg/day for extra 3 months. Urinary iodine concentration was measured by arsenic/cerium catalyzing spectrophotography. In addition, the early apoptosis was assessed by flow cytometry with Annexin V-FITC staining, while the late apoptosis was evaluated by TUNEL microscopy. Furthermore, the rate of cell death and cell cycle kinetics were determined by flow cytometry with propidium iodide (PI) staining. Moreover, the level of intracellular reactive oxygen species (ROS) quantified by the fluorescence intensity was measured via flow cytometry with a DCFH-DA probe. Immunohistochemistry technique was also used for the detection of the expression of Bcl-2 and Bax. RESULTS: The apoptotic rate and ROS production in thyrocytes were significantly elevated in Groups 2 and 3 after 4 and 8 months of treatment compared to the control group ( P<0.05). Groups 2 and 3 were also found to have greatly higher percentage of cells in S phase after 4 and 8 months of exposure (5%-6% vs. 3%, P<0.05), with a significant reduction in cells in G0G1-phase (64% 67% vs. 80%, P<0.05). These differences disappeared after normal iodine intake for extra 3 months. The expressions of Bcl-2 and Bax among different groups remained unchanged. Positive correlations were observed between the apoptotic rate, ROS level and urinary iodine concentration after 4 and 8 months of excessive iodine exposure (r is in the range of 0.736~0.855, all of Ps<0.01). CONCLUSION: The experimental data indicated that more apoptosis of thyrocytes after exposure to excessive iodine was attributed to the increased free radical production, other than regulated Bcl-2 and Bax expression. The effects of iodine excess are reversible. PMID- 20013612 TI - Fas ligand (FasL, Apo-1L/CD95L) expression and clinical outcome in papillary microcarcinoma and papillary thyroid carcinomas with diameter smaller than 1.5 centimeters. AB - CONTEXT: The presence and the importance of FasL (FasL, APO-1L/CD95L) in papillary microcarcinoma lesions which are smaller than 1.5 cm of the thyroid is unclear. OBJECTIVE: It was aimed to investigate FasL expression in papillary microcarcinoma (PMC) of the thyroid. DESIGN: FasL immunoreactivity was evaluated in PMC lesions. Paraffin sections of thyroid specimens obtained from 59 papillary thyroid carcinoma consecutive patients were stained using antibody to FasL. Fas ligand expression and the relation and comparison with clinical and pathological findings in PMC were determined. RESULTS: There were 39 females (66.1%) and 20 males (33.9%) aged 23-74 years (51.9+/-11.8 yrs), and 20 patients with tumor size <=5 mm, 24 patients with 6-10 mm and 15 patients with 10-15 mm. The mean of the intensity and the percentage of FasL immunoreactivity were significantly higher in lesions of PMC than peripheral thyroid tissue (29.5+/-37.9% and 1.42+/-1.25 vs. 2.1+/-5.4% and 0.46+/-0.95, respectively; p<0.001). Fas ligand immunoreaction were not different according to tumor size, the presence of tumor capsule and tumor invasion, invasion of thyroid capsule, peripheral tissue and vascular structures and multicentricity (p>0.05). However, FasL positivity, staining and intensity were high in patients above 45 years, in oncocytic and tall cell variants, in TNM stage pT4A (p<0.05). CONCLUSIONS: This study showed that FasL expression (positivity, staining and intensity) was high and increased in PMC of the thyroid tissue, and above 45 years, in tall cell and oncocytic variants, and in advanced tumor. PMID- 20013613 TI - [Cocaine-related gastric perforation]. AB - Since the 1980s the abuse of cocaine has been -associated with gastroduodenal perforations in the United States. Here, we report the case of a 28-year-old man who came to our hospital with severe abdominal pain after smoking cocaine. Physical examination revealed generalised abdominal guarding. His X-ray did not show any free intraperitoneal air. However, there was a slightly elevated white blood cell count. Upon laparoscopic exploration of the abdomen, the -patient was found to have a generalised peritonitis secondary to a perforation of the prepyloric anterior wall. The operative procedure consisted of ulcer excision and primary closure with a pyloroplasty as well as an extensive abdominal irrigation after laparotomy. PMID- 20013614 TI - Oral administration of the NMDA receptor antagonist S-ketamine as add-on therapy of depression: a case series. PMID- 20013615 TI - Desynchronization: a novel model to induce heart failure. AB - BACKGROUND: Previous large animal heart failure models led to inhomogeneous results. Therefore, we developed a novel model combining rapid pacing with forced ventricular desynchronization. METHODS: Heart failure was induced in 20 pigs during a pacing period of 21 days. Group A (n = 10) received one right ventricular lead (220 bpm). In group B (n = 10), two leads were implanted in different right ventricular regions with beat-to-beat alternation of activation sites (each lead 110 bpm). Sham-operated pigs (n = 6) served as controls. Hemodynamics were invasively evaluated and tissue was analyzed by immunohistochemistry and zymography. RESULTS: Hemodynamics were significantly more impaired in group B with an increase of pulmonary capillary wedge and central venous pressure and a reduction of cardiac index (control 4.3 +/- 0.1 l/min/m (2); A 3.6 +/- 0.2; B 2.9 +/- 0.2, P < 0.05). Heart-to-body weight ratio was significantly higher in group B. Histological analyses showed a significant increase of cell diameters and interstitial fibrosis with significantly higher collagen contents in group B. CONCLUSION: The new model with a combination of rapid pacing and forced desynchronization of the ventricular contraction is superior to traditional heart failure models induced solely by rapid pacing. PMID- 20013616 TI - Extracorporeal life support - systems, indications, and limitations. AB - Extracorporeal life support is becoming increasingly popular with the miniaturization of the devices necessary for cardiac and pulmonary support. In the present report, the current systems, indications for placement and limitations are reviewed. PMID- 20013617 TI - Aprotinin-associated risks in off-pump coronary artery bypass grafting. AB - BACKGROUND: Little data is available regarding the safety of using the serine protease inhibitor aprotinin in off-pump cardiac surgery. We retrospectively assessed the risks of administering the drug to adult patients undergoing off pump coronary artery bypass grafting (OPCABG). METHODS: Aprotinin was administered as a bolus of 1 or 2 million kallikrein inhibiting units to 391 patients following median sternotomy; 370 control patients underwent surgery during the same time period without receiving aprotinin. No other antifibrinolytic agents were administered. RESULTS: Preoperative characteristics, length of ICU and hospital stay were similar between the mostly medium-risk aprotinin and the control patients. Postoperative cardiac, renal, neurological, and respiratory complications and hospital mortality occurred with comparable frequencies in both groups. Levels of myocardial enzymes during the first 72 h after surgery also did not differ significantly. CONCLUSION: Use of aprotinin in OPCABG was not associated with a higher incidence of hospital mortality, cardiovascular, renal, or other complications. Given the good safety profile in this large patient population we suggest that aprotinin could still be a valid antifibrinolytic treatment option in OPCABG. PMID- 20013618 TI - The impact of pre- and postoperative renal dysfunction on outcome of patients undergoing coronary artery bypass grafting (CABG). AB - OBJECTIVE: Acute changes in renal function after elective coronary bypass surgery represent a challenging clinical problem. In this study, we evaluated perioperative risk factors for the development of postoperative renal dysfunction (PRD), and the impact of such an event on the perioperative course. Additionally, we investigated the influence of preoperatively mildly increased serum creatinine on perioperative mortality and morbidity. METHODS: We retrospectively analyzed data of 2511 patients undergoing isolated CABG between 2004 and 2007 with a preoperative serum creatinine < or = 2.2 mg/dL. There were 592 patients with a preoperative serum creatinine of between 1.4 and 2.2 mg/dl (mild renal dysfunction group) and 1919 patients with a serum creatinine < 1.4 mg/dl. Perioperative risk factors for PRD were analyzed by multivariate regression analysis. RESULTS: Global in-hospital mortality was 3.1 %.The incidence of PRD was 6.2 %. Mortality for patients who had PRD was 7.8 vs. 2.9 % for patients who did not ( P < 0.05). PRD increased the length of hospital stay by 3.7 days (12.2 vs. 15.9; P < 0.05). Multivariate logistic regression identified the following variables as independent predictors of PRD: age, angina class III/IV, diabetes mellitus, prolonged cardiopulmonary bypass time, and preoperative serum creatinine. With regard to preoperative renal function, we found that operative mortality was higher in the mild renal dysfunction group (5.7 % vs. 2.5 %; P < 0.05). New dialysis/hemofiltration (5.1 % vs. 1.2 %; P < 0.05) and postoperative stroke (5.1 % vs. 1.6 %; P < 0.05) were also more common in these patients. CONCLUSIONS: Mild renal dysfunction preoperatively is an important predictor of outcome after CABG. In these patients, PRD dramatically increases mortality, morbidity and length of hospital stay. PMID- 20013619 TI - Postoperative treatment of carvedilol following low dose landiolol has preventive effect for atrial fibrillation after coronary artery bypass grafting. AB - BACKGROUND: Postoperative atrial fibrillation (AF) is the most common complication after coronary artery bypass grafting (CABG). We have reported that the intra- and perioperative administration of landiolol has a preventive effect on postoperative AF in the early postoperative period after CABG surgery. The purpose of this study was to investigate the prophylactic effect of postoperative treatment with carvedilol following landiolol against postoperative AF. METHODS: We reviewed all patients who underwent CABG from December 2005 and February 2009. Fifty-three patients underwent scheduled isolated CABG and were divided two groups; carvedilol group (n = 31), and control group (n = 22). Incidences of postoperative AF were noted. RESULTS: There was no statistical difference between the two groups with regard to the occurrence of AF after CABG. The maximum ventricular rate of AF was significantly lower in the carvedilol group than in the control group. All patients with AF in the control group needed treatment for tachycardia, but this was not necessary in the carvedilol group. CONCLUSIONS: Postoperative treatment with carvedilol following landiolol has the effect of preventing tachycardia during AF after coronary artery bypass grafting. PMID- 20013620 TI - Sternum fractures and effects of associated injuries. AB - BACKGROUND: Aim of this study was to determine the clinical characteristics, morbidity and mortality in patients with a sternum fracture (SF). METHODS: From October 1998 to December 2008, 80 patients with sternal fractures were admitted to the Thoracic Surgery Department of the University Hospital. The records of all patients were reviewed retrospectively. Patients' age and gender, extent of sternal and thoracic injury, types of associated injuries, treatment and outcome, the length of hospital stay, morbidity and mortality were collected. We divided our patients into two groups: Group I with an isolated sternum fracture and Group II with a sternum fracture and additional injury. RESULTS: Patients' ages ranged from 18 to 83 years with an average age of 48.7 +/- 15.4 years. There were 67 male (84 %) and 13 female (16 %) patients. Thirty-five patients (44 %) sustained an isolated SF and the remaining 45 (56 %) had a SF in combination with associated injuries. The commonest etiology of the SF was a motor vehicle accident (77 %). Most of the fractures were located at the sternal body (76 %). The most common associated injuries were rib fractures (30 %), followed by extremity injuries (18 %). The great majority of patients (81 %) was treated with conservative measures such as bed rest, analgesia, etc. The remainder was treated either by tube thoracostomy (15 %), thoracotomy (1.25 %) or sternal fixation (2.5 %). There was no mortality and morbidity in Group I, but the morbidity and mortality rates of Group II were 15 % and 1 %, respectively. Mean length of hospital stay for all the patients was 5.7 days. CONCLUSION: Isolated SFs are benign entities that can be safely managed with rest and analgesia. Associated injuries in patients with SF determine the morbidity and length of hospital stay. PMID- 20013621 TI - Early recurrence after surgical resection in patients with pathological stage I non-small cell lung cancer. AB - INTRODUCTION: Early recurrence is observed even in patients who undergo complete resection and had pathological (p-) stage I. Therefore, we focused on early recurrence, and attempted to elucidate the relationship between early recurrence and clinicopathological factors. METHODS: Between May 1993 and December 2005, 1201 patients with non-small cell lung cancer (NSCLC) underwent surgical treatment at our institution. Of these, 402 patients who underwent complete resection and had p-stage I NSCLC were retrospectively analyzed for clinicopathological factors. Patients were divided into four groups according to the period between surgery and recurrence (R): no recurrence (NR, n = 331), late recurrence (LR, n = 28, R > 2 years), intermediate recurrence (IR, n = 22, 1 year < R < or = 2 years), and early recurrence (ER, n = 21, R < or = 1 year). RESULTS: The overall 5-year survival rate for patients with p-stage I was 79.9 %. The overall 5-year survival rates were 91.0 %, 55.6 %, 17.1 %, and 7.5 % for the NR, LR, IR, and ER group, respectively. Preoperative high CEA level, lymphatic permeation, and pleural invasion were proven to be independent factors for overall recurrence. Moreover, multivariate analysis showed that preoperative CEA level, pathological T factor, lymphatic permeation, vascular invasion, and pleural invasion influenced early recurrence within one year. CONCLUSIONS: The present study demonstrated that preoperative CEA level, pathological T-factor, lymphatic permeation, vascular invasion, and pleural invasion were independent prognostic factors for early recurrence within one year, even in patients with pathological stage I. In patients with these factors, adjuvant therapy may be indicated since this may improve their survival. PMID- 20013622 TI - Autologous blood pleurodesis for persistent air leak. AB - OBJECTIVE: Air leak is a well-known problem, often leading to great discomfort and prolonged hospitalization. Many methods have been proposed for the prevention or treatment of prolonged air leak, but none of them turned out to offer the perfect solution. The purpose of this study was to evaluate the results of blood pleurodesis in patients with persistent air leak. MATERIAL AND METHOD: From February 2005 through March 2007, 20 patients (14 male and 6 female) ranging in age from 41 to 81 years underwent blood pleurodesis in our department due to persistent air leak (> 7 days). In the majority of patients the underlying disease was emphysema (n = 14). Lobectomy was performed in 10 cases, LVRS in 4 and 3 patients suffered from secondary pneumothorax. Autologous blood pleurodesis was performed at bedside. With apical chest tube placement and an expanded lung, 60 ml of blood was taken from the femoral vein and instilled into the thoracic cavity. Because of the high air leakage the drainage was not clamped but was positioned over the patient to prevent blood running out of the pleural space, but allowing air to be evacuated. Patients were asked to rotate in bed every 15 minutes over a period of 6 hours. The tube was removed 12 h after cessation of the air leak. RESULTS: In 14 patients (70 %) closure of the fistula was achieved in less than 12 hours, in 3 within 24 h, in 2 within 48 h, while one patient with LVRS and NSCLC needed reoperation. Only in 2 cases was a second instillation of blood performed. Over a 2- to 24-month follow-up period neither complications nor recurrences were observed. CONCLUSION: The injection of autologous blood into the pleural space through an existing chest tube is an easy-to-perform, painless and inexpensive method with a high success rate. PMID- 20013623 TI - Surgical resection of chest wall tuberculosis. AB - BACKGROUND: Chest wall tuberculosis is a rare disease. After reviewing cases previously treated by surgical resection, the methods of resection, results, perioperative complications, and recurrence of the disease were analyzed. METHODS: The clinical and radiological data of 21 patients with chest wall tuberculosis treated between March 1998 and May 2007 were reviewed retrospectively. RESULTS: Symptoms included growing chest wall mass, with the time intervals of surgical resection from symptoms ranging from one to eight months (mean 2.3 months). Fourteen patients had a past history of tuberculosis. Preoperative needle aspiration was performed in 10 patients. The lesion was confined to the chest wall without a pleural lesion in 4 patients; 14 patients had a chest wall lesion with a pleural space. Resection of the abscess without rib resection was performed in nine patients. Complete resection of the abscess together with soft tissue and the involved rib was performed in 12 patients. Recurrence occurred in two patients (9.5 %): one underwent complete resection with rib resection while the other had resection without rib resection. CONCLUSION: Chest wall tuberculosis requires surgical resection in most cases and complete surgical resection may be needed to keep the recurrence rate low. PMID- 20013624 TI - Prognosis of patients after pulmonary artery plasty for non-small cell lung cancer. AB - OBJECTIVE: We evaluated the clinical outcomes of patients after lung resection with pulmonary artery (PA) plasty for non-small cell lung cancer (NSCLC). METHODS: From 1995 to 2006, 36 patients (26 males and 10 females) with NSCLC underwent lobectomy or segmentectomy with PA plasty at our institution. The mean age of the patients was 65.9 years old (range 45-87 years old). There were 17 left upper lobectomies, 10 right upper lobectomies, five left lower lobectomies, two right upper-and-middle bilobectomies, one right lower lobectomy, and one left upper division segmentectomy. Both bronchoplasty and PA plasty were performed in 15 patients. Six patients received preoperative chemotherapy, and one had preoperative radiotherapy. RESULTS: The postoperative morbidity rate was 27.8 % (10/36), and the mortality rate (30 days) was 2.8 % (1/36). One patient underwent completion pneumonectomy on postoperative day 13. Macroscopic residual cancer was identified in two patients at the thoracic wall and aorta, respectively; microscopic residual cancers were identified in two patients at the stumps of the pulmonary artery and in one patient at the bronchial stump. Postoperative radiation therapy was additionally given to those four patients, except one. The 5-year survival rate for all patients was 51.8 %. There was no significant difference in the 5-year survival rate between clinical N (cN) 0-1 patients and cN2 patients. However, in pathological N (pN) 0-1 patients, the 5-year survival rate was significantly better than that of pN2 patients (71.9 % versus 0.0 %; P < 0.001). CONCLUSIONS: PA plasty for NSCLC is acceptable and highly recommended for pN0-1 patients. Strict patient selection should be considered so as to avoid surgical operations in patients with pN2 staging. PMID- 20013625 TI - Vena cava resection with venous-venous shunt for hemangioma. AB - Mediastinal hemangiomas are rare neoplasms among mediastinal tumors. We report a rare case of hemangioma of the vena cava that was detected incidentally in a 30 year-old female patient. PMID- 20013626 TI - Pneumopericardium and pneumomediastinum as a late complication of defibrillator implantation after coronary artery bypass graft surgery. AB - A 69-year-old male underwent implantation of a cardioverter-defibrillator with cardiac resynchronization therapy (CRT) for symptomatic ventricular tachyarrhythmia (VT) and severe left ventricular (LV) dysfunction with an ejection fraction (EF) of 30 % and dyssynchrony via a left subclavian venous access. Twenty days after the procedure, the patient complained of shortness of breath and was found to have a 30 % apical left pneumothorax on chest X-ray as a not unusual complication of the subclavian venous access. A computed axial tomography of the chest revealed pneumopericardium and associated pneumomediastinum as a complication of the CRT implantation and persisting microscopic pleuro-pericardial fistula as a consequence of previous coronary artery bypass graft surgery (CABG), accidentally diagnosed three years after the procedure. The pneumothorax and pneumopericardium were small and did not require chest tube placement. The patient was treated conservatively and his subsequent course was excellent. PMID- 20013627 TI - Successful surgical treatment of giant main coronary artery fistula connecting to right atrium. AB - Coronary artery fistulas are rare; they are defined as congenital or acquired coronary artery abnormalities in which blood is shunted into a cardiac chamber, great vessel, or other structure, bypassing the myocardial capillary network. We report here on a case of successful surgical treatment of a giant left main coronary artery fistula in a 29-year-old man who presented to our institution with exertional dyspnea. In this report, the different methods of management are also discussed and the indications for the surgical technique are briefly outlined. PMID- 20013628 TI - Long-term survival of a patient with type A thymoma and Masaoka's stage IV b: case report. AB - Thymoma is the most common neoplasm in the anterior mediastinum. This report presents an extremely rare case of a type A thymoma with Masaoka's stage IV b due to lymph node metastasis. In 1997, a 59-year-old man underwent complete resection of a stage IV b type A thymoma with postoperative radiotherapy to the mediastinum. In 2006, small nodules were detected in the anterior mediastinum and above the right diaphragm and tumor resection was performed. The two lesions were both histologically diagnosed as recurrences of the type A thymoma. There has been no evidence of recurrence 15 months after the second surgery. PMID- 20013629 TI - A pulmonary sclerosing hemagioma with an increasing uptake on PET. AB - Pulmonary sclerosing hemangioma is a relatively rare neoplasm of the lung. Although there have been five previous case reports that address the 18-fluoro-2 deoxyglucose positron emission tomography (FDG-PET) findings in a sclerosing hemangioma of the lung, no report has demonstrated an interval change in the FDG PET findings. This report describes a case of pulmonary sclerosing hemangioma which presented with an increase of uptake of FDG-PET after about one years' follow-up. PMID- 20013630 TI - Sternoclavicular joint infection: a case report. AB - Infection of the sternoclavicular joint (SJI) is a rare problem accounting for approximately 1 % of cases of septic arthritis. Patients typically present with symptoms of localized pain lasting a period of several weeks with or without systemic signs of fever and chills. Confirmation is made by aspirating the joint, and broad spectrum antibiotics should be tailored to treat the identified organisms. SJI can be treated conservatively with intravenous antibiotics and repeat imaging, but surgical intervention is required if patients present with an abscess, osteomyelitis or mediastinitis. PMID- 20013631 TI - Simultaneous pulmonary embolectomy and aortic root replacement. AB - Massive pulmonary embolism (PE) is characterized by hypotension and cardiogenic shock due to right ventricular failure, and is associated with a high mortality rate. In this case study, we report a simultaneous pulmonary embolectomy and aortic root replacement in a 71-year-old woman with a known ascending aortic aneurysm who sustained a massive PE following a VATS left upper lobectomy for non small cell lung cancer. PMID- 20013633 TI - [Adaptation of a wireless PC mouse for modification of GUI during intervention in an open highfield MRI at 1.0T]. AB - PURPOSE: To demagnetize a PC mouse for interventional use in magnetic resonance imaging (MRI). MATERIALS AND METHODS: A mobile version of a wireless optical mouse at 2.470 Hz was modified in ways ranging from the replacement of magnetic components to the construction of new components. The magnetic force (Fm), the induction of susceptibility artifacts, and the signal-to-noise ratio (SNR) were determined. The modified mice at 1.0 T. were used in MR-guided interventions. RESULTS: A final demagnetization down to 2 % (Fm = 9 N vs. Fm = 0.15 N) was achieved. Operating the device clinically caused no compromise in image quality. From a minimal distance of about 20 cm, even the non-modified mouse did not produce any susceptibility artifacts in gradient recalled imaging. The SNR was not reduced significantly (TSE 228 + or - 22,3 vs. 211 + or - 15; FFE 55 + or - 5,6 vs. 56 + or - 4,3). For over a year the modified mouse proved to be a reliable tool for the interventionalist to manipulate MRI. CONCLUSION: Relatively inexpensive modifications of a standard USB mouse enable the interventionalist to work independently without a technical assistant and to take immediate advantage of multiplanar MR imaging during interventions. PMID- 20013634 TI - Ponderal index of the newborn: effect of smoking on the index of the small-for gestational-age infant. AB - The ponderal index (PI) is evaluated in the context of its distribution within a given population. Low PI (<10th percentile for gestational age) has been extensively studied but not much is known about the distribution and factors associated with a high (>90th percentile) PI among small-for-gestational-age (SGA) infants. This retrospective study of singleton live first births from 1990 to 2007 in a Midwestern city explores factors associated with a high PI, particularly among SGA infants. Independent variables included exposures (none, smoke, hypertension) and maternal and infant demographic characteristics. There were 45,453 births, 28 to 42 weeks' gestational age, 55% Whites and 51% male. Mean PI increased with gestational age and was highest among Hispanics and lower among SGA infants. High PI was present in 11% of appropriate-for-gestational-age and 4% of SGA infants. Among SGA infants, odds ratios (ORs) of high PI were higher for smoke exposure (1.21; 95% confidence interval 0.97, 1.87) and lower for males (0.66; 0.47, 0.93). In conclusion, the distribution of PI varies by exposures and of high PI by race/ethnicity/gender. SGA infants with high PI have relative surplus of mass, and ostensibly, adiposity, for their frame. There is a need to use PI in exploring and defining previously observed associations between SGA and adult-onset obesity/metabolic syndrome. PMID- 20013635 TI - Isoflavonoids from the roots of Campylotropis hirtella. AB - From the dried roots of Campylotropis hirtella (Franch.) Schindl., five novel isoflavonoids, 3( S)-7,2',4'-trihydroxy-5,5'-dimethoxy-6-(3-methylbut-2-enyl) isoflavan ( 1), 3( S)-2',4'-dihydroxy-5,5'-dimethoxy-(6'',6''-dimethylpyrano) (2'',3'':7,6)-isoflavan ( 2), 3( R)-5,4'-dihydroxy-2'-methoxy-3'-(3-methylbut-2 enyl)-(6'',6''-dimethylpyrano)-(7,6 : 2'',3'')-isoflavanone ( 3), 3( R)-5,4' dihydroxy-2'-methoxy-(6'',6''-dimethylpyrano)-(7,6: 2'',3'')-isoflavanone ( 4), and 3( R)-5,4'-dihydroxy-7,2'-dimethoxy-6-geranylisoflavanone ( 5) were isolated. The structures of the compounds were elucidated on the basis of spectroscopic analysis. All isolates exhibited good immunosuppressive activities on mitogen induced splenocyte proliferation, and their cytotoxicity on splenic lymphocytes was also tested. PMID- 20013636 TI - Determination of flavonoids in licorice using acid hydrolysis and reversed-phase HPLC and evaluation of the chemical quality of cultivated licorice. AB - Licorice contains flavonoids and triterpenoids as the major bioactive components. Most of the flavonoids are the glycosidic form of liquiritigenin (LIQ), isoliquiritigenin (ISO) and formononetin (FOR). A reversed-phase HPLC method for the quantification of LIQ, ISO and FOR in licorice was developed. This method does not measure each glycoside but measures the aglycones using acid hydrolysis. All calibration curves showed good linear regression (r > 0.9998). The method showed good precision for intraday (RSD < 2.14 %) and interday (RSD < 0.51 %) assays. The limit of detection was 0.031 microg for LIQ, 0.011 microg for ISO and 0.006 microg for FOR. The limit of quantification was 0.31 microg for LIQ, 0.11 microg for ISO and 0.06 microg for FOR. The flavonoid contents along with the glycyrrhizin content of cultivated licorice from seedling plants in Japan and commercial wild licorice were investigated. This new method could be extremely useful for evaluating the quality of licorice. PMID- 20013637 TI - Toxicity assessment of nine types of decoction pieces from the daughter root of Aconitum carmichaeli (Fuzi) based on the chemical analysis of their diester diterpenoid alkaloids. AB - Various processed types of FUZI (the daughter roots of the highly toxic plant Aconitum carmichaeli Debx, FZ) decoction pieces (the herbal materials processed according to the specifications of Chinese medicine manuals; " YINPIAN" in Chinese transliteration) are widely used in traditional medicine to treat various diseases, but their toxicities are not known. Nine types of FZ decoction pieces, including one raw slice and eight processed forms, were therefore prepared, each in 7 to 10 batches, to assess for their toxicity. Altogether 84 FZ samples were quantified on the amount of highly toxic diester diterpenoid alkaloids, i.e., aconitine, mesaconitine and hypaconitine by a newly developed HPLC method with HPLC-DAD and LC-MS techniques. The comparison of the processed FZ to raw slices of the root showed that the amount of each analyte in the processed FZ was drastically decreased. The sum of the three toxic compounds in the 8 types of processed FZ was only 3.91-34.80 % of this value in the FZ raw slice. This implies that the toxicity of processed FZ was decreased significantly. The amounts of toxic components in the 8 types of processed FZ varied significantly, often by a power of ten, indicating that the dosage of these herbs, when prescribed for clinical uses, should be cautiously set in order to avoid poisoning incidents. PMID- 20013638 TI - Schistosomiasis suppressing deoxyphorbol esters from Euphorbia cauducifolia L. latex. AB - The molluscicidal activity of E. Cauducifolia L. latex, extracted in various organic solvents, was tested against Biomphalaria glabrata snails, using Bayluscide as a control. The ethyl acetate extract was found to be the most active and in bioassay guided HPLC fractionation yielded eight ( 1- 8) compounds. The structure and relative configuration of the isolates were established through spectroscopic (UV, IR, (1)H, (13)C NMR, 2D NMR, HSQC, HMQC, HMBC, COSY-45 degrees , TOCSY, HOHAHA, HOESY, ROESY, NOESY, SECSY, and NOE) techniques and mass measurements. These were named as: 13-acetoxy-20- O-angeloyl-12-deoxyphorbol ( 1), 13- O-[N-(2-aminobenzoyl)]anthraniloyl-20-acetoxy-12-deoxyphorbol ( 2), 13,20 O-dibezoyl-12-deoxyphorbol ( 3), 13,20- O-diangeloyl-12-deoxyphorbol ( 4), 13- O angeloyl-20- O-[N-(2-aminobenzoyl)]anthraniloyl-12-deoxyphorbol ( 5), 13- O tigloyl-20- O-[N-(2-aminobenzoyl)]anthraniloyl-12-deoxyphorbol ( 6), 13- O benzoyl-20- O-[N-(2-aminobenzoyl)]anthraniloyl-12-deoxyphorbol ( 7), and 13- O hexanoyl-20- O-[N-(2-aminobenzoyl)]anthraniloyl-12-deoxyphorbol ( 8). The literature reveals that all of the isolates were new natural metabolites and active against mollusks. Compounds 1 and 2, which were esterified at C-13 with acetoxy or N-(2-aminobenzoyl) anthraniloyloxy, showed twice the activity of the control while others ( 3- 8) were equipotent. PMID- 20013640 TI - [Announcement -- Doctoral prize of the German Lung Foundation]. PMID- 20013639 TI - Anti-HIV-1 diterpenoids from leaves and twigs of Polyalthia sclerophylla. AB - Bioassay-guided fractionation and purification of the anti-HIV-1-active MeOH extract from the leaves and twigs of Polyalthia sclerophylla led to the isolation of two new compounds, ENT-kaur-sclerodimer ( 1) and cyclotucanol 3-palmitate ( 2), along with the known ENT-kaur-16-en-19-oic acid ( 3), 15 beta-hydroxy- ENT kaur-16-en-19-oic acid ( 4), 15 beta-acetoxy- ENT-kaur-16-en-19-oic acid ( 5), 15 oxo- ENT-kaur-16-en-19-oic acid ( 6), 16 alpha,17-dihydroxy- ENT-kauran-19-oic acid ( 7), 16 alpha-hydroxy- ENT-kauran-19-oic acid (xylopic acid) ( 8), a pseudodimer (15 alpha-hydroxy- ENT-kaur-16-en-19-oic acid/17-hydroxy- ENT-kaur-15 en-19-oic acid) ( 9), ermanin, nicotiflorin, and allantoin. Among these isolates, compound 3 was the most active in both anti-syncytium (EC (50) 13.7 microg/mL and selectivity index 3.1) and HIV-1 reverse transcriptase (IC (50) 34.1 microg/mL) assays. PMID- 20013641 TI - [Sleep medicine -- monitoring sleep apnea at home]. PMID- 20013642 TI - [Not Available]. PMID- 20013643 TI - [Not Available]. PMID- 20013644 TI - [Announcement -- Award of the DGP]. PMID- 20013645 TI - [Chronic lung disease -- genetics for bronchial hypersecretion discovered]. PMID- 20013646 TI - Animal models of primary aldosteronism. AB - Primary aldosteronism is considered the most prevalent form of secondary hypertension with pathophysiological and clinical features different from those of essential hypertension. Despite its high prevalence with the exception of the small subgroup of patients with familial hyperaldosteronism type I, the underlying genetic and molecular basis of this common disease is still largely unknown. In this context animal models can provide important insights in the physiology of aldosterone regulation that can serve as a starting point for investigation of mechanisms involved also in autonomous aldosterone secretion. Mouse models with defined genetic modification can further be utilized to prove functional relevance of these predefined candidate genes.Finally, animal models can be used to investigate cardiovascular and metabolic consequences of unopposed aldosterone secretion and potential restoration of these parameters through pharmacologic interventions. This review will provide a brief overview on animal models currently available for primary aldosteronism and describe in vivo screening strategies that are likely to aid in the elucidation of molecular and genetic mechanisms involved in autonomous aldosterone secretion. PMID- 20013647 TI - Fibroblast growth factor 19 serum levels: relation to renal function and metabolic parameters. AB - Fibroblast growth factor 19 (FGF19) was recently introduced as a novel metabolic regulator reversing diabetes mellitus, hepatic steatosis, hyperlipidemia, and adiposity. In the current study, we determined circulating FGF19 levels in patients on chronic hemodialysis (CD) as compared to controls with a glomerular filtration rate (GFR) above 50 ml/min. FGF19 was measured by ELISA in control (n=60) and CD (n=60) patients and correlated to clinical and biochemical measures of renal function, glucose, and lipid metabolism, as well as inflammation, in both groups. Median serum FGF19 levels were 1.5-fold higher in CD patients (266.7 microg/l) as compared to subjects with a GFR above 50 ml/min (178.1 microg/l) (p=0.001). Furthermore, fasting glucose negatively and independently predicted circulating FGF19 in controls (p<0.05). Moreover, adiponectin was a positive and C-reactive protein was a negative independent predictor of FGF19 serum concentrations in CD patients. Taken together, we have demonstrated that circulating FGF19 levels are significantly increased in end-stage renal disease. Furthermore, FGF19 is associated with a beneficial metabolic profile in both control and CD patients. PMID- 20013648 TI - Reduction in Size of a Large Rathke's Cleft Cyst on Treatment with Low Dose of Corticosteroid. AB - Rathke's cleft cyst is a non-neoplastic sellar cyst, which is increasingly reported on radiological investigation performed for unrelated intra-cranial pathology. When symptomatic, it is associated with headache, visual symptoms, and pituitary dysfunction. We report a case of an 18 year-old male patient with Rathke's cleft cyst, who presented with failing vision, headache, and hypocortisolism. After defaulting on planned surgery, the patient continued to take a replacement dose of prednisolone for a year. He reported significant improvement in vision and remarkable reduction in cyst size on repeat imaging after a year. Surgery was later performed in view of persisting severe headache. The authors discuss the reduction in cyst size in relation to long-term usage of replacement steroid. They postulate that selected patients with Rathke's cleft cyst with radiological evidence of inflammatory fluid can be given a trial of glucocorticoids and assessed for cyst shrinkage and changes in imaging characteristics. PMID- 20013649 TI - C-11 methionine positron emission tomography/computed tomography localizes parathyroid adenomas in primary hyperparathyroidism. AB - In patients with primary hyperparathyroidism (pHPT), positive preoperative localization studies enable to perform a minimally invasive approach for parathyroid surgery. However, current imaging techniques are not always successful. We therefore conducted a study to determine the sensitivity of C-11 methionine positron emission tomography/computed tomography (Met-PET/CT) in localizing parathyroid adenomas in pHPT. Met-PET/CT scans of the neck and mediastinum of 33 patients undergoing parathyroidectomy for primary HPT were compared with intraoperative and histological findings. Primary HPT was caused by a single gland adenoma in 30 patients, while another 3 patients had multiglandular disease. Met-PET/CT scan correctly located a single gland adenoma in 25 out of 30 (83%) patients with pHPT, among them 2 patients with persistent disease, 7 patients with prior neck surgery, and 8 patients with concomitant thyroid nodules. In 3 patients with multiglandular disease, Met-PET/CT showed only one enlarged parathyroid gland in two individuals and was negative in the third patient. Statistical analysis found a significant correlation between true positive results and the weight (2.42+/-4.05 g) and diameter (2.0+/-1.18 cm) of parathyroid adenomas while the subgroup with false negative findings had significantly smaller (0.98+/-0.54 cm) and lighter (0.5+/-0.38 g) glands. Sensitivity was 83% for single gland adenomas and 67% for multiglandular disease. Met-PET/CT correctly localized 83% of single gland parathyroid adenomas in patients with pHPT. However, preoperative localization of multiglandular disease due to double adenomas or parathyroid hyperplasia remained difficult. PMID- 20013650 TI - Factors engaged in reactivation of DNA replication in the nuclei of growing mouse oocytes introduced into the cytoplasm of parthenogenetic one-cell embryos. AB - Mammalian primary oocytes are arrested in the post-replicative G2 phase of the cell cycle. In contrast to other G2 nuclei, the nucleus of the growing mouse oocyte can reinitiate DNA synthesis after transfer by cell fusion under favorable cytoplasmic conditions, created by the parthenogenetic one-cell embryo. In the present study, we used the cell hybrid system to analyze the distribution of proteins involved in DNA re-replication in the oocyte nucleus. We show that this process is preceded by an extensive rearrangement of the insoluble fractions of minichromosome maintenance (MCM) proteins (Mcm2, -6 and 7). We also demonstrate that Cdc6 protein is present in primary growing mouse oocytes freshly isolated from the ovary, in a soluble and insoluble form. In contrast to MCM proteins, the insoluble fraction of Cdc6 was not rearranged in oocyte nuclei reinitiating DNA replication in hybrid cells. The rearrangement of MCM proteins and reinitiation of DNA synthesis occurred in the nuclei, in which the nuclear envelope remained intact. Reinitiation of DNA replication in the oocyte nucleus was sensitive to the inhibition of both CDK activity and polyadenylation of maternal mRNAs, indicating a role of proteins synthesized de novo by the embryo. These results allow us to understand better the mechanisms involved in the reinitiation of DNA replication in growing oocytes. PMID- 20013651 TI - Zygotic VegT is required for Xenopus paraxial mesoderm formation and is regulated by Nodal signaling and Eomesodermin. AB - The T-box gene VegT plays a crucial role during mesendoderm specification of the amphibian embryo. While the function of maternal VegT (mVegT) has been extensively investigated, little is known about the function and transcriptional regulation of zygotic VegT (zVegT). In the present study, we used comparative genomics and a knockdown experiment to demonstrate that zVegT is the orthologous gene of zebrafish Spadetail/Tbx16 and chick Tbx6L/Tbx6, and has an essential role in paraxial mesodermal formation. zVegT knockdown embryos show several defects in the patterning of trunk mesoderm, such as abnormal segmentation of somites, a reduction in muscle, and the formation of an abnormal mass of cells at the tail tip. We also identified the cis-regulatory elements of zVegT that are necessary and sufficient for mesoderm-specific expression. These cis-regulatory elements are located in two separate upstream regions of zVegT, corresponding to the first intron of mVegT. The results of in vitro binding and functional assays indicate that Forkhead box H1 (FoxH1) and Eomesodermin (Eomes) are the trans-acting factors required for zVegT expression. Our results highlight the essential role of zVegT in organization of paraxial mesoderm, and reveal that zVegT is regulated by a coherent feedforward loop of Nodal signaling via Eomes. PMID- 20013652 TI - Competition for ligands between FGFR1 and FGFR4 regulates Xenopus neural development. AB - Cell-surface-localized receptors and their extracellular ligands usually comprise distinct families and promote diversity of signal transduction regulation. The number of available ligand molecules is often the limiting factor for receptor activation during interpretation of the signal by the responding cell. Limited ligand availability in a particular area of tissue should lead to local competition between different members of a receptor family for binding and subsequent activation. Fibroblast growth factor receptor (FGFR) 4 (FGFR4) is a less potent activator of downstream pathways than FGFR1, the major subtype of FGFR. Regional expression of Xenopus FGFR1 and FGFR4 (XFGFR1 and XFGFR4, respectively) overlap in the anterior part of prospective and developing neural tissue. In this paper we show that XFGFR1 and XFGFR4 have opposing effects on the positioning of expression domains of mid- and hindbrain markers when the expression levels of the receptors are altered. We present a line of evidence to support our hypothesis that competition between XFGFR1 and XFGFR4 for ligands is required for correct positioning of marker expression. Local competition between receptors with different potencies should provide an efficient means for a cell to interpret the ligand signal correctly, and may constitute a more general mechanism for regulating signal transduction. PMID- 20013653 TI - Pluripotency of bank vole embryonic cells depends on FGF2 and activin A signaling pathways. AB - The objective of this study was to investigate the capability of bank vole (Myodes glareolus) embryonic cells to sustain their pluripotent character during in vitro culture, and to determine the optimal conditions for derivation of embryonic stem (ES) cells. We compared the presence of specific pluripotency (Oct4, Ssea1) and differentiation markers (Gata4 - primitive endoderm marker; Cdx2 - trophectoderm marker) in blastocysts and inner cell mass (ICM) outgrowths obtained from blastocysts of bank vole, and two mouse hybrids F1(C57Bl/6xCBA/H) and F1(C57Bl/6x129/Sv), which differ in the permissiveness of giving rise to ES cells. We found that, in contrast to mouse, the expression of pluripotency markers in the cells of bank vole ICM outgrowths is progressively downregulated and rapidly lost by the 4th day of culture. This correlates with the appearance of cells expressing Gata4 and Cdx2, indicating differentiation towards primitive endoderm and derivatives of trophectoderm, respectively. We have also shown that heterologous cytokine leukaemia inhibitory factor (LIF) in conjunction with either homologous or heterologous feeder layer is unable to delay differentiation and preserve pluripotency of bank vole embryonic cells. Thus, the conditions optimised for mouse do not support the maintenance of bank vole embryonic cells in the undifferentiated state and do not allow for the isolation of the ES cells. Instead, combination of fibroblast growth factor 2 and activin A allows retention of Oct4 expression in bank vole blastocyst outgrowths during 4-day culture, indicating that signaling pathways operating in human, rather than mouse ES cells, might be involved in the process of self-renewal of bank vole embryonic cells. PMID- 20013654 TI - Over-expression of thymosin beta 4 promotes abnormal tooth development and stimulation of hair growth. AB - Thymosin beta 4 has multi-functional roles in cell physiology. It accelerates wound healing, hair growth and angiogenesis, and increases laminin-5 expression in corneal epithelium. Furthermore, thymosin beta 4 stimulates tumor growth and metastasis by induction of cell migration and vascular endothelial growth factor mediated angiogenesis. Using a construct on the skin-specific keratin-5 promoter, we have developed thymosin beta 4 over-expressing transgenic mice to further study its functional roles. Thymosin beta 4 in adult skin and in embryonic stages of the transgenic mouse was analyzed by both Western blot and immunohistochemistry. The over-expression of thymosin beta 4 was observed especially around hair follicles and in the teeth in the transgenic mice. We examined the phenotype of the thymosin beta 4 over-expressing mice. Hair growth was accelerated. In addition, the transgenic mice had abnormally-shaped white teeth and dull incisors. We found that the expression of laminin-5 was up regulated in the skin of the transgenic mice. We conclude that thymosin beta 4 has an important physiological role in hair growth and in tooth development. PMID- 20013655 TI - Signalling molecules involved in mouse bladder smooth muscle cellular differentiation. AB - Mouse bladder mesenchyme differentiates into smooth muscle under the influence of urothelium at gestational day 13.5 (E13.5). Sonic hedgehog (Shh) is considered to be the upstream gene arising from the urothelium, which induces smooth muscle in the peripheral bladder mesenchyme. We hypothesize differential gene expression across the full thickness of bladder mesenchyme as a function of proximity to the inducing bladder urothelium and the peripheral location of the smooth muscle. Embryonic bladders from FVB mice were collected at E12.5, 13.5, 15 and 16 and cryosectioned followed by microdissection with a PixCell II laser capture microscope. RNA extraction was performed at the laser captured sites and mRNA expression profiles were measured using SYBR Green quantitative RT-PCR. Smooth muscle a-actin (SMAA) and smooth muscle myosin heavy chain (SM-MHC) were expressed in the E13.5, E15 and E16 bladders in the peripheral layer of mesenchyme, but not in the prospective submucosa. Patched 1 (Ptc1), Gli1 and bone morphogenetic protein (Bmp) 4 expression was consistently elevated in the mesenchymal layer immediately adjacent to the urothelium compared to the peripheral location at E12.5. After E12.5, Ptc1 expression decreased to an undetectable level throughout the bladder mesenchyme. The level of TGF-beta1 was highest in the mesenchymal layer adjacent to the serosa at E13.5. The level of expression of serum response factor (SRF) was also highest at E15 in the peripheral mesenchyme. Genes downstream of Shh are differentially expressed in the prospective submucosa vs. the peripheral bladder mesenchyme as a function gestation age and smooth muscle differentiation. PMID- 20013656 TI - Diallel analysis of leaf disease resistance in inbred Brazilian popcorn cultivars. AB - We estimated general and specific combining abilities and examined resistance to northern leaf blight (Exserohilum turcicum) and to gray leaf spot (Cercospora zeae-maydis) in a set of nine inbred popcorn lines. These inbreds were crossed in a complete diallel scheme without reciprocals, which produced 36 F(1) hybrids. Two experiments with a square lattice design and three replications were conducted during the 2008/2009 crop season, in Maringa, PR, Brazil. The severity of northern leaf blight and gray leaf spot was assessed under natural infestation conditions. Data were examined by individual and joint analysis of variance. Individual and joint Griffing's diallel analyses were carried out for adjusted means. General combining ability and specific combining ability were significant (P < 0.10) by the F-test for northern leaf blight and gray leaf spot infestation levels. This denotes that additive and non-additive gene effects both contributed to resistance to these diseases, but that the additive gene effects were more important. Among the inbred lines, P(8) and P(9) gave the highest resistance to northern leaf blight, and P(3) and P(4.3) gave the highest resistance to gray leaf spot. The hybrids P(7.4) x P(8) and P(4.3) x P(9) could be exploited by reciprocal recurrent selection to provide genotypes with both northern leaf blight and gray leaf spot resistance. Significant interaction between general combining ability and crop season (P < 0.10) denotes the importance of environment, even though the disease levels in the hybrids were quite consistent. PMID- 20013658 TI - Meiotic behavior of a nonaploid accession endorses x = 6 for Brachiaria humidicola (Poaceae). AB - Brachiaria humidicola (Poaceae), originally from Africa, is an economically important pasture plant in tropical South America. An accession of B. humidicola (H038) collected from the wild African savanna (Mbeya, Tanzania) showed irregular microsporogenesis. This meiotic behavior was consistent with an allopolyploid origin. Multivalent chromosome association at diakinesis gave tri- to octavalents, associated with two nucleoli in some cells. Six non-congregated univalents in metaphase I and anaphase I, along with previous lines of evidence for x = 6 in B. humidicola, confirm H038 as a nonaploid accession, 2n = 9x = 54. Asynchrony in the genome during microsporogenesis also corroborated this assumption. Its putative origin could be a cross between two related species with different rhythms in meiosis. The meiotic behavior of this accession reinforces the hypothesis of the existence of a new basic chromosome number (x = 6) for Brachiaria. The use of this accession in the breeding of this important forage grass for the tropics is discussed. PMID- 20013657 TI - HPV31 and HPV33 incidence in cervical samples from women in Recife, Brazil. AB - Human papillomavirus (HPV) has been extensively studied concerning genomic structure, infection mechanisms, and diversity of types, as well as disease progression stages and development of vaccines. HPV type prevalence can differ in specific populations in different countries, according to ethnicity. This is the first report of an integrated project to evaluate the incidence of HPV types in different regions in Brazil in order to obtain data for vaccine development. Cervical samples were collected from women seen at a public hospital in Pernambuco, Northeast Brazil, for routine evaluation of genital alterations. Selection of the patients was random. There was a strong prevalence of HPV16 and a high incidence of HPV types 31 and 33. These data foster the discussion about the need to evaluate viral prevalence in each geographic region in order to develop targeted vaccine programs. PMID- 20013659 TI - XRCC1 polymorphisms and haplotypes in Mexican patients with acute lymphoblastic leukemia. AB - We examined the influence of the Arg194Trp, Arg280His, and Arg399Gln polymorphisms of XRCC1 (X-ray repair cross-complementing group 1) on the development of childhood acute lymphoblastic leukemia (ALL) in 120 ALL patients and 120 controls in Mexico. All of them were genotyped for these polymorphisms, using polymerase chain reaction. No significant differences in allele and genotype frequencies for any polymorphism were observed between patients and controls. Estimation of haplotypes showed the eight expected haplotypes (A-H), seven of which were found in both patients and controls; haplotype A (Arg-Arg Arg) was the most common, whereas haplotypes F and G were absent in patients and controls, respectively. Haplotype B (Trp-Arg-Arg) was found to be associated with an increased risk of ALL (odds ratio (OR) = 1.95, 95% confidence interval (CI) = 1.13-3.37; P = 0.016), particularly in males (OR = 2.65, 95%CI = 1.25-5.63; P = 0.01). Individually, the 194Trp, 280His, and 399Gln alleles were not associated with significantly increased risk for ALL in these Mexican children. PMID- 20013660 TI - Estimation of genetic distance between 10 maize accessions with varying response to different levels of soil moisture. AB - Ten maize accessions (NC-9, A50-2, M-14, B-42, NC-3, T-7, N-48-1, B-34, USSR, and WFTMS) were studied to estimate the genetic distance on molecular level by random amplified polymorphic DNA. These accessions were selected on the basis of their variable responses against different levels of moisture. Twenty-five primers were used to test genetic diversity, of which 14 were observed to be polymorphic. Ninety-three loci were amplified; among these, 77 showed polymorphism and the other 16 were monomorphic. Primers A-13 and C-02 gave the most polymorphic bands, while primers A-01 and C-06 gave the fewest polymorphic bands. The genetic similarities of the 10 maize accessions ranged from 82.8 to 54.8%. Accessions USSR and WFTMS showed greatest similarity, and accessions M-14 and B-42 were found more dissimilar than the other accessions. On the basis of cluster analysis, these 10 accessions were classified in two major groups, A and B, and than further divided into sub-groups. The cluster analysis showed that accessions in the same group as well as in the sub-groups were similar in their physical and morphological characters, since the characters are controlled genetically. PMID- 20013661 TI - RNA-binding motif protein RBM22 is required for normal development of zebrafish embryos. AB - RBM22 is a newly discovered RNA-binding motif protein, belonging to the SLT11 family; it has been reported to be involved in pre-splicesome assembly and to interact with the Ca(2+) -signaling protein ALG-2. However, previous studies have not demonstrated whether its expression is essential for early embryogenesis in vertebrates. We utilized zebrafish as a developmental model to study the role of RBM22 during embryogenesis. The aligned protein sequences of RBM22 were found to be highly conserved. In situ analysis showed that zRBM22 was expressed as early as the one-cell stage, implying maternal origin during oogenesis; embryos continued to express zRBM22 through at least the 32-h post-fertilization (hpf) stage of development. RT-PCR analysis confirmed the in situ expression pattern, indicating that RBM22 is expressed without any spatial and temporal specificity during zebrafish development. zRBM22 morpholino-induced alterations in development were observed as early as 15 hpf of embryogenesis, based on a morphological analysis. Embryogenesis was monitored through 32 hpf; knockdown resulted in a truncated axis and developmental arrest of the head and tail. We conclude that RBM22 plays an important role in zebrafish embryogenesis. PMID- 20013662 TI - Vagal paraganglioma of the neck: a case report. AB - Vagal paragangliomas are rare in the head and neck. Complementary use of computed tomography and magnetic resonance imaging can facilitate the diagnosis and help determine the best management approach. Most paragangliomas should be treated with surgery. We report a case of vagal paraganglioma of the neck in a 50-year old man. The patient was treated with superficial parotidectomy via a transcervical approach. No postoperative morbidity was noted, and at 3 years of follow-up, he was free of disease. PMID- 20013663 TI - Acquired toxoplasmosis of the buccal area with extranodular involvement: report of an unusual case. AB - Acquired toxoplasmosis is a common parasitic infection in humans. It can be caused by ingestion of infected meat or other food that has been contaminated by the feces of infected cats. Approximately 90% of immunocompetent patients with acquired toxoplasmosis are asymptomatic and undiagnosed; in the other 10%, toxoplasmosis manifests as a nonspecific, self-limited illness that usually does not require treatment. In symptomatic cases, cervical lymphadenopathy is one of the most common clinical findings. We report the case of a 33-year-old woman who experienced unilateral facial swelling secondary to toxoplasmosis. In addition to the atypical location of her disease (i.e., the buccal area), the atypical histopathologic findings in this case (e.g., extranodular involvement) constituted a very unusual presentation of toxoplasmosis. PMID- 20013664 TI - A large, persistent, solitary vellus hair cyst in the postauricular area. AB - Vellus hair cysts typically appear as numerous small papules in the precordial area in children. Treatment is generally not necessary as most cases resolve spontaneously. We report a case of a vellus hair cyst that was unique in that it manifested as a solitary mass in the postauricular area and it had persisted for 16 years. The tumor was surgically excised. We also review the literature on this condition. PMID- 20013665 TI - Clinical significance of a hearing and imaging workup in a child with cochlear nerve aplasia and a normal internal auditory canal. AB - Modern magnetic resonance imaging has significantly improved the diagnosis of cochlear nerve deficiencies. A careful assessment of all clinical, imaging, and auditory data is of utmost importance in such cases in order to properly establish the site of the abnormality. We report the case of a 3-year-old girl with unilateral cochlear nerve aplasia, normal middle and inner ear anatomy, and an absence of otoacoustic emissions, all of which erroneously suggested cochlear damage. We also briefly review the embryogenesis of the inner ear and auditory pathway. A patient with similar findings in the setting of a bilateral hearing loss and insufficient imaging would be at risk of inappropriate cochlear implantation. PMID- 20013666 TI - Review of inactive ingredients in antibiotic ear drops. AB - The authors review the relevant chemical and biological characteristics of inactive ingredients in antibiotic ear drop drug products. Although many of the older aminoglycoside-containing otic products are still used today, only some fluoroquinolone-containing ear drops, approved in the last decade, have been proven safe and effective according to current rigorous scientific standards, including ototoxicity testing of the formulations. Preservatives, antioxidants, surfactants, buffers, suspending and emulsifying agents, osmotic agents, viscosity modifiers, solvents, and penetration enhancers are reviewed with regard to their functional role in formulations, as well as their potential for ototoxicity. PMID- 20013667 TI - Use of the laryngeal microdebrider blade in treating Teflon-induced vocal fold granuloma: a case report. AB - Vocal fold granuloma induced by polytef (Teflon) injection is rare. This condition requires surgical intervention in order to restore the voice. In this report, we discuss the use of an endoscopic laryngeal microdebrider blade in successfully treating this condition. To the best of our knowledge, this procedure has not been previously described in the literature. PMID- 20013668 TI - Carcinoma of the prostate presenting as a painful parotid mass with mandibular invasion: a case report. AB - Prostate cancer metastatic to the parotid gland is exceedingly rare, as only 10 cases have been previously reported in the literature. Symptoms may mimic a parotid infection or suggest a primary parotid tumor. We report a new case of carcinoma of the prostate metastatic to the parotid. The tumor was painful and had invaded the mandible. Fine-needle aspiration of the mass and immunohistochemical staining for prostate-specific antigen confirmed the diagnosis. The patient died 1 month later of an unrelated cause. PMID- 20013669 TI - Adenoid cystic carcinoma of the trachea metastatic to the nasal cavity: a case report. AB - Cases of carcinoma metastatic to the nasal cavity are rare. We report the case of a 63-year-old woman with a metastasis to the nasal cavity from a primary tracheal adenoid cystic carcinoma (ACC). The nasal tumor was treated with surgical resection. No evidence of any local recurrence was observed at 4 years of follow up. To the best of our knowledge, no case of a tracheal ACC metastatic to the nasal cavity has been previously reported in the literature. Although rare, metastatic disease to the nasal cavity should be considered in patients who have a known primary carcinoma elsewhere and who present with nasal symptoms. PMID- 20013670 TI - Legal considerations with a joint hearing aid venture. PMID- 20013671 TI - A giant vocal fold polyp causing dyspnea. PMID- 20013672 TI - Nasal myiasis: the role of endoscopy. PMID- 20013673 TI - Clinical manifestations of sinonasal undifferentiated carcinoma. PMID- 20013674 TI - The postauricular island flap (flip-flop flap). PMID- 20013675 TI - Eosinophilic esophagitis. PMID- 20013676 TI - Revision stapedectomy. PMID- 20013677 TI - Necrotizing otitis externa: a new trend? Report of 6 atypical cases. AB - Necrotizing otitis externa (OE) is traditionally seen in elderly diabetics and immunocompromised patients. During a 7-year period at our institution, we treated 6 patients with necrotizing OE who were not diabetic, who were not immunocompromised, and who were relatively young (age range: 27 to 65 yr; mean: 42.8). The necrotizing OE in these patients was less aggressive but just as severe as its usual presentation in older diabetic or immunocompromised patients. All 6 patients had evidence of bony erosion on computed tomography of the temporal bones, although 4 had negative findings on technetium-99m scintigraphy. Four of the 6 patients required mastoid exploration and fascia lata grafting, and the other 2 chose to undergo extensive daily microdebridements and intravenous antibiotics followed by 6 weeks of oral antibiotics. We recommend that a diagnosis of necrotizing OE be kept in mind when evaluating any patient who presents with severe otalgia, particularly in the presence of Pseudomonas aeruginosa infection of the external auditory canal, edema, granulation tissue, and bony erosion. PMID- 20013680 TI - Pediatric airway consultation survey in a tertiary care children's hospital: an interobserver analysis. AB - We conducted a prospective analysis to independently observe the initial opinion of: (1) consulting pediatric residents (postgraduate year 2 [PGY-2]), (2) corresponding consulting attending physicians (general pediatric or pediatric emergency medicine), and (3) consulting otolaryngology PGY-2 residents when assessing patients for "noisy breathing." Each was asked to define the type of noisy breathing present and to formulate a diagnosis based on a limited set of choices. The final diagnosis was determined by a single attending pediatric otolaryngologist (S.E.S.) at the completion of the diagnostic workup. The accuracy for characterization of breath sounds for the pediatric residents, attending pediatricians, and otolaryngology residents were 26.1%, 23.5%, and 98.6%, respectively. The positive predictive values for the determination of diagnosis were 71.6%, 69.4%, and 76.6%, respectively. These findings indicate that pediatric residents and attending pediatricians may be deficient in their ability to characterize the breath sounds commonly seen in patients with noisy breathing. Further education of pediatricians may lead to a more accurate diagnostic evaluation of a child with noisy breathing. PMID- 20013679 TI - An asymptomatic schwannoma of the nasal septum: report of a unique case. AB - Schwannomas of the nasal septum are extremely rare, and therefore their diagnosis and treatment can pose certain challenges. We describe the case of an incidentally discovered schwannoma of the nasal septum that was unique in that the lesion was completely asymptomatic. In view of our finding, we believe that routine examinations of both nasal fossae should be considered during routine diagnostic laryngeal flexible endoscopy. PMID- 20013681 TI - Primary sinonasal malignant melanoma: a clinicopathologic and prognostic study. AB - We conducted a retrospective study to evaluate the clinicopathologic features and prognostic factors associated with primary sinonasal malignant melanoma. We reviewed the records of 10 patients-7 men and 3 women, aged 35 to 70 years (mean: 51.4)-who had been treated at our referral center over a 9-year period. The tumors were located in the nasal cavity in 7 patients and in a paranasal sinus in the other 3. Medial maxillectomy was performed in 4 patients, wide local excision in 3, and endoscopic excision in 2; the remaining patient, who had presented with a liver metastasis, received chemotherapy and palliative local radiotherapy. Two patients who presented with a neck node metastasis also underwent concurrent radical neck dissection. Follow-up ranged from 8 to 70 months (median: 25). Only 4 patients remained alive through the duration of follow-up. The 2- and 5-year survival rates were 60 and 40%, respectively. Based on the findings of our small study, we conclude that primary sinonasal malignant melanoma carries a generally poor prognosis despite aggressive treatment. The primary cause of death in our series was a distant metastasis (n = 5) despite adequate locoregional control in most cases. This finding confirms the aggressive nature of this disease. Other factors that appeared to be associated with a poor prognosis were (1) older age, (2) a primary tumor location in a paranasal sinus, (3) an advanced tumor stage, (4) an external approach to surgery, and (5) the absence of adjuvant radiotherapy. PMID- 20013682 TI - [Heart failure I: Clinical aspects, causes. diagnostic strategies]. PMID- 20013683 TI - [CME--Laboratory 14]. PMID- 20013684 TI - [What is your diagnosis? Pronounced benign pneumatosis intestinalis in the framework of COPD]. PMID- 20013685 TI - [What is your diagnosis? Pyoderma gangrenosum]. PMID- 20013686 TI - [From pay-for-effort to pay-for-performance--an analysis of the Swiss health care system with focus on the inpatient sector]. AB - INTRODUCTION: The Swiss health care system is facing the implementation of lump compensation in the form of diagnosis related groups from 2010 on. In addition there is an increasing discussion about the quality of health care in the media. We have analyzed current remuneration in Swiss health care and their steering effects on providers in order to deduct future developments in Swiss health care remuneration. METHODS: Based on the remuneration contracts and tariff regularities at the Basel University Hospital we conducted an internet and literature search. The identified Swiss remuneration systems were classified after remuneration scales and remuneration item using a typology of performance related remuneration systems. The steering effects of the remuneration systems on the providers were deducted. RESULTS: Remuneration scales can be classified in <>, <> or <>. Remuneration items can be classified in <>, <> or <>. Remuneration systems can lead to increased or decreased services or to patient selection. In the Swiss health care system we find a trend away from traditional <> to <> remuneration systems. In that context diagnosis related groups are identified as an intermediate step in the development of remuneration systems. CONCLUSIONS: Future developments of medical remuneration in terms of a consideration of quality of medical performance and negotiated costs seem likely in Switzerland in the long term. Both, economics and quality should be considered adequately in a health care remuneration system. PMID- 20013687 TI - [Autosomal dominant polycystic kidney disease (ADPKD): rapamycin as a new treatment option]. AB - Autosomal dominant polycystic kidney disease (ADPKD) is characterized by the massive enlargement of both kidneys caused by numerous cysts. The cyst formation begins in utero and the continuous growth of cysts leads to compression and destruction of non-cystic renal parenchyma, so that finally a kidney replacement in the 5th to 6th decade of life is necessary. So far there is no therapy that halts disease progression. Animal data show that pharmacological inhibition of a central regulator of cell proliferation, the so-called mammalian target of rapamycin (mTOR), slows disease progression. This overview provides an insight into the disease and the specific mTOR inhibitor sirolimus, which is currently tested in clinical trials. PMID- 20013688 TI - [Urological treatment of renal calculi]. AB - As a disease, nephrolithiasis can not only be exceedingly bothersome for the respective patients, it is also very expensive for our public health system. Indications for an active treatment of renal calculi go mainly according to the stone size as well as the clinical symptoms. The goal of modern nephrolithiasis treatment is the complete removal of all stones from the pelvi-calyceal system with the lowest possible morbidity. Extracorporeal shockwave lithotripsy, percutaneous nephrolitholapaxy and flexible ureterorenoscopy are the main options for treatment. According to the stonesize, -localisation and -composition these options are given different priorities. However, due to significant technical progress in the field of endourology there is a clear trend towards flexible ureterorenoscopy, which is a promising modality for the treatment of kidney stones for a number of reasons. PMID- 20013689 TI - [Non-invasive techniques for the diagnosis of melanoma]. AB - In the last years a number of new non invasive techniques for the early diagnosis of melanoma have become very popular. In addition to dermoscopy, total body photography and digital dermoscopy frequently assist the dermatologist in differentiating nevi from early melanomas. A new promising technique for the non invasive diagnosis of melanoma might be confocal microscopy. PMID- 20013690 TI - [Slow ventricular tachycardia]. PMID- 20013691 TI - [Diarrhoea, nausea and vomiting]. AB - We saw a 54-year-old patient who had been treated for gastrointestinal ulcerations with bleeding complications now presenting with nausea, vomiting and diarrhoea. The patient has been suffering from recurrent diarrhea for years. The suspicion of a neuroendocrine tumor had been made but could not be confirmed intraoperatively in the past. As suspicion was still high with high levels of chromogranin A and high gastrin levels the diagnosis now could be confirmed with a somatostatin scintigraphy. Finally the confirmed spot was identified to be a metastasis of a gastrinoma of still unknown primary localisation. PMID- 20013692 TI - [Quadrivalent HPV vaccination for women 24 to 45 years old]. PMID- 20013693 TI - [Beginning of a nicotine substitute chewing gum before smoking cessation has to be finished is not more effective than at the start of the term]. PMID- 20013696 TI - [Physicians -- paintings puzzle]. PMID- 20013698 TI - [Management of COPD by chest physicians in Poland]. PMID- 20013699 TI - [Leptin serum levels in cachectic and non-cachectic lung cancer patients]. AB - INTRODUCTION: Leptin, protein taking part in body mass regulation, might play a role in cancer cachexia development. The aim of the study was to measure leptin serum levels in cachectic, non-cachectic lung cancer patients, healthy controls and to correlate leptin concentration with nutritional status markers. MATERIAL AND METHODS: 40 lung cancer patients were enrolled into the study: 20 with cachexia, 20 without cachexia, and 10 healthy controls. Leptin serum concentration, body mass, BMI, arm circumference and skin triceps fold thickness were measured in each subject. RESULTS: Serum leptin level in cachectic cancer patients was significantly lower than in non-cachectic and healthy controls. Leptin concentration correlated with body mass, arm circumference and skin triceps fold thickness. CONCLUSIONS: Cachectic lung cancer patients have significantly lower serum leptin concentrations than non-cachectic patients and healthy controls which may suggest, that leptin does not play an important role in cancer cachexia development. Leptin levels positively correlate with good nutritional status markers. Non-cachectic lung cancer patients have similar leptin serum levels as healthy controls. PMID- 20013700 TI - [Outpatients specialist care of chronic obstructive pulmonary disease patients in Poland - results of the KOMPAS study]. AB - INTRODUCTION: COPD is one of the most important lung diseases. It is responsible for significant proportion of outpatients pulmonary clinics visits. Data on outpatients care of COPD patients in Poland are limited. This article presents design of the KOMPAS study and basic results relating to COPD patients. MATERIAL AND METHODS: The aim of this prospective study was to describe population of patients with COPD treated by specialists, and to characterize methods of care used in various COPD stages. Participating physicians used pocket PCs equipped with specially developed software to collect COPD patients' data. RESULTS: Data on 2958 COPD patients were analyzed. 2/3 of patients were males. Mean patients age was 63 years and mean FEV(1) - 57% of predicted value. 86% of all subjects underwent spirometry during the first visit or the previous 6 months. Cough and exertional dyspnoea were the most frequently reported COPD symptoms (about 80% of patients). At the first visit 32% of subjects were current smokers and 51% were ex-smokers. 17% of all study patients have never smoked. Before inclusion 2/3 of patients were treated with bronchodilatator, and about 1/3 with inhaled steroid. After entering the study, over 90% received bronchodilatator and more than 2/3 received inhaled steroid. Differences in treatment between stages of COPD based on its severity were relatively small. CONCLUSIONS: Results of the KOMPAS study provide basic information about COPD outpatients care in Poland. These data, especially concerning physicians' compliance with current COPD guidelines, may be useful in planning undergraduate/postgraduate training for physicians and for those who are responsible for health resource allocation. PMID- 20013701 TI - [Diagnostic utility of the molecular assay GenoType MTBC (HAIN Lifesciences, Germany) for identification of tuberculous mycobacteria]. AB - INTRODUCTION: The GenoType system (HAIN Lifescience, Germany) offers new perspectives of detecting the tuberculous and non-tuberculous mycobacteria at the molecular level. The system compromises five independent tests that could be performed either on direct specimens or isolated strains, to identify the strains and test the resistance against rifampin and isoniazid. Up to now, non GenoType test was applied in Poland. The aim of the study was an evaluation the accuracy of GenoType MTBC test in speciation of the clinical isolates, previously classified as M. tuberculosis complex by HPLC analyze of mycolic acids. MATERIAL AND METHODS: 161 clinical isolates, derived from the TB patients hospitalized in the Warsaw Medical University Hospital between 1999 and 2007 were assayed. RESULTS: On the basis of the hybridization patterns, all 161 studied strains were identified as M. tuberculosis/M. canettii. CONCLUSIONS: 1. The GenoType MTBC test (HAIN Lifescience, Germany) precisely recognizes M. tuberculosis complex. The 100% accordance in speciation of M. tuberculosis by the GenoType MTBC test as compared to HPLC method was demonstrated. The GenoType MTBC test can replace HPLC in detection of tuberculous mycobacteria in clinical isolates. 2. As the GenoType MTBC test performs well, the other tests of GenoType system may be considered to be verified in diagnostic procedure of mycobacterial infection. PMID- 20013702 TI - [Program of early detection of pulmonary neoplasms by the computed tomography - preliminary Szczecin experience]. AB - INTRODUCTION: Lung cancer (LC) remains one of the most serious epidemiological and clinical challenges both in the world and Poland. Results of LC therapy are far from satisfaction. One of the reasons of high LC mortality is its late detection. Currently, few centers in the world conduct LC screening programs based on low-dose spiral computed tomography (CT) of the chest. There have been no such programs in Poland up to date. MATERIAL AND METHODS: The program of LC early detection based on CT for citizens of Szczecin aged 55-65, who smoked at least 20 pack/years, was introduced on May 1st 2008 and was planned for 3 years. There were 3647 subjects examined till December 31st 2008. Algorithm of further action for detected lesions was based on the IELCAP and NELSON trial protocols. RESULTS: There were 25 malignancies detected, including 21 LC (17 females and 4 males) up to date (70% were in stage I TNM). In contrast - there was only 16.8% stage IA LC detected in the comparable group diagnosed on the symptoms basis. Fifty seven patients were treated surgically, of whom 16 underwent lobectomy or pneumonectomy coupled with radical mediastinal lymphadenectomy. There were 3 wedge resections and 2 segmentectomies performed, too. Perioperative mortality was 0%. There were 32 benign lesions of different clinical importance resected as well (tuberculoma, hamartoma, inflammatory, mycotic and sarcoidal lesions). In our group 1365 lesions were detected in 996 persons - they are followed up in accordance with the IELCAP algorithm. CONCLUSIONS: Early LC detection program initiated in Szczecin resulted in significant increase of stage IA TNM detected patients subsequently treated radically. There was also a large number of small non malignant lesions detected. PMID- 20013703 TI - [Dendritic cells and allergy]. AB - Dendritic cells (DC) are pivotal regulator of immune reactivity and immune tolerance. The aim of this paper is to present the current knowledge on the dendritic cell development, biology and their role in asthma and immunotherapy. PMID- 20013704 TI - [Disorders with elevated immunoglobulin E levels]. AB - Immunoglobulin E (IgE) was first described in 1967. It has the lowest serum concentration of all circulating immunoglobulin isotypes. Serum IgE concentration at birth is low and rises until the age of 10-15 years. The increment of serum IgE in children with predisposition to atopic reactions is usually more abrupt. Total serum IgE levels decline from the second to the eight decade of life. Usually, elevated IgE concentration is related to atopic diseases but there are many other disorders combined with elevated serum IgE (for example: infections, neoplasms, immunodeficiency syndromes, skin disorders, inflammatory diseases). The spectrum of diseases combined with serum IgE elevation was presented in the present article. PMID- 20013705 TI - [Chlamydial infection in asthma. Macrolide treatment]. AB - Atypical microorganism infections, including Chlamydophila pneumoniae, play an important role in asthma course. A significant influence of chlamydial infection on severity of asthma exacerbations and increase in chronic asthma symptoms has been shown. The group of medication with high antibacterial activity against atypical microorganisms are macrolides, which also have anti-inflammatory and immunomodulatory effects. Macrolide treatment in patients with asthma can be connected with additional therapeutic benefits. Mechanism of action of these antibiotics is not ultimately clarified and further studies are required. PMID- 20013706 TI - [Vascular endothelial growth factor - structure, function and role in airways inflammation and the clinical course of asthma]. AB - Vascular endothelial growth factor (VEGF) is produced by a wide range of cells and plays an important role in normal and pathological angiogenesis. The formation of the vascular system is essential for inflammatory process in airways, especially in the course of chronic lung diseases - for example bronchial asthma. Increased level of VEGF in serum and induced sputum has been demonstrated in patients with stable asthma, as well as during the exacerbation of this disease. VEGF end VEGF-receptor's expressions is closely correlated with neovascularization and seems to be an important predictor of the lung's remodeling. Due to direct causative relationship between inflammatory process and angiogenesis in airways, it was suggested, that inhibiting VEGF's activity would have an anti-inflammatory effect on airways. In this article the potential role of vascular endothelial growth factor in airways inflammation, remodeling and the clinical course of bronchial asthma is presented. PMID- 20013707 TI - [The impact of psychological variables on the presentation and progress of asthma and patient's cognitive functions]. AB - Chronic respiratory system diseases become serious public health problem all over the world. The most prevalent are obstructive diseases (asthma and COPD). The prevalence of asthma is still high and concern patients representing wide range of age and socio-economic status. Despite progress in diagnostic and therapeutic options several studies showed that asthma has an impact on health-related quality of life and patients' coping. Asthma as chronic condition results in limitations of patients activity and social relations. Thus psychosocial variables, which may have an impact on asthma symptoms presentation and disease progress, should be considered. There are only few reports concerning cognitive functions in asthma. The aim of the study was to assess the potential impact of psychosocial factors on asthma symptoms presentation, and cognitive function in asthma patients. PMID- 20013708 TI - [Chest ultrasound in the diagnosis of pulmonary embolism in a pregnant patient - a case report]. AB - Pregnancy is a risk factor for both pulmonary embolism (PE), and an incorrect diagnostic assessment in cases of suspected PE with potentially dangerous consequences for the mother and foetus. The major concern is ionising radiation utilized by diagnostic tests and its potential negative effect on foetal safety. This paper presents diagnostic difficulties encountered in a 31-year-old patient at 20 weeks of gestation who was admitted to hospital with non-specific chest pain and suspected PE as a complication of right lower limb venous thrombosis. The case study reminds of chest ultrasound as a useful tool in the diagnosis of PE. The official clinical practice guidelines do not recommend the use of chest ultrasound for diagnosing of PE due to lack of a sufficient number of published studies. This case report may encourage further, prospective studies in the hope to define whether and when chest ultrasound might find its place in the diagnostic strategy of PE, especially in pregnant women. PMID- 20013709 TI - [Dr Maria Wiktoria Werkenthin - a memory on the 65th anniversary of her death]. PMID- 20013710 TI - [In memory of Maria Werkenthin]. PMID- 20013711 TI - [Commentary to the article of T. Grodzki et al. "Program of early detection of pulmonary neoplasms by the computed tomography - preliminary Szczecin experience"]. PMID- 20013712 TI - [A robotic system for gait re-education in patients with an incomplete spinal cord injury]. AB - INTRODUCTION: A spinal cord injury involves the loss or alteration of motor patterns in walking, the recovery of which depends partly on the rearrangement of the preserved neural circuits. AIM. To evaluate the changes that take place in the gait of patients with incomplete spinal cord injuries who were treated with a robotic walking system in association with conventional therapy. PATIENTS AND METHODS: The study conducted was an open-label, prospective, descriptive trial with statistical inference in patients with C2-L3 spinal cord injuries that were classified as degrees C and D according to the American Spinal Injury Association (ASIA) scale. The variables that were analysed on the first and the last day of the study were: number of walkers, 10-m gait test, the Walking Index for Spinal Cord Injury scale revision, technical aids, muscle balance in the lower limbs, locomotor subscale of the measure of functional independence, modified Ashworth scale for spasticity and the visual analogue scale for pain. At the end, data were recorded from the impression of change scale. The analysis was conducted by means of Student's t, chi squared and Pearson's correlation; p < or = 0.05. RESULTS: Forty-five patients, with a mean age of 44 +/- 14.3 years, finished the study; 76% were males, injury was caused by trauma in 58% of cases, and the time of progression was 139 +/- 70 days. Statistically significant increases were observed in the number of subjects capable of walking, walking speed, less need for technical aids, strength in the lower limbs and independence in activities of daily living. CONCLUSIONS: Treatment using the robotic system in association with conventional therapy improves walking capacity in patients with incomplete spinal cord injuries. PMID- 20013713 TI - [Memory deficit in patients with subcortical vascular cognitive impairment versus Alzheimer-type dementia: the sensitivity of the 'word list' subtest on the Wechsler Memory Scale-III]. AB - INTRODUCTION: Memory deficits are frequent in mild subcortical vascular pathology and in the early stage of Alzheimer's disease. AIM: To study the memory deficits in patients with subcortical vascular cognitive impairment (SVCI) vs. mild stage Alzheimer's disease patients (AD), using the Weschler Memory Scale-III (WMS-III) word lists test, to examine the adequacy of this test to show the different memory patterns in this population, that could contribute to early differential diagnosis. PATIENTS AND METHODS: Three groups of patients were compared: AD (n = 25), SVCI-leukoaraiosis (n = 17) and SVCI-multi infarct (n = 16). The three groups did not differ in age, education or severity of illness. RESULTS: Patients with SVCI showed a memory impairment in free recall with an improvement in their performance on the recognition task. The AD group, however, showed low scores in free recall as in recognition tasks, with a major number of false positive errors. Significant differences were also found between the SVCI-leukoaraiosis and SVCI-multi infarct groups, with the latter showing the best performance in long term retention and a minor trend to respond with false positive errors. CONCLUSIONS: The WMS-III word lists test is a good instrument to differentiate the memory profile between SVCI and AD, with the recognition task being the most discriminative one. The worst impairment in leukoaraiosis patients in comparison to the multi infarct group, suggests that white matter diffuse lesion could affect more directly the recall processes mediatised by the frontal lobe. PMID- 20013714 TI - [The role of platybasia in trigeminal neuralgia]. AB - INTRODUCTION: The vascular compression of a nerve is known to be the most important cause of idiopathic trigeminal neuralgia (ITN) by most authors. However, several cases of skull base deformities in patients with ITN have been reported in our bibliographic review. AIM. To compare the existence of platybasia between a group of patients with ITN and a randomized control group. SUBJECTS AND METHODS: Twenty-five patients with ITN and twenty-five control subjects were X rayed and their basal angles were measured and compared. RESULTS: The basal angle measured to investigate the existence of platybasia is significantly greater in the ITN group than in the control group. Platybasia was found in 10 patients with ITN. However, only two cases of platybasia were reported in the control group. Therefore in our study we have found that the incidence of platybasia in patients with ITN is significantly increased. CONCLUSION: Our data demonstrated that platybasia has an important role in the pathogenesis of the ITN by altering the normal anatomy of the nerves and vessels within the posterior fossa. PMID- 20013715 TI - [A clinico-anatomical correlation study of logorrhoea]. AB - INTRODUCTION: The degree of verbal production necessary to be considered logorrhoea has still not been defined, and no clear correlation has been established between the topography of the dysfunction and this symptom. AIMS: To provide quantitative data about normal verbal production and to identify the location within the brain of the alterations observed in neuroimage of patients with logorrhoea. SUBJECTS AND METHODS: The oral verbal production of 60 control subjects between 20 and 80 years of age was quantified by analysing five speeches. Ten patients who exceeded the 75th percentile in at least two of the five speeches underwent structural and functional neuroimaging tests. RESULTS. The data on verbal production of normal subjects are reported. Age, sex and habits (smoking, coffee, alcoholic drinks) did not exert an influence, but the degree of schooling was seen to have an effect. All the patients were diagnosed with frontotemporal degeneration, although in one case there were also coexisting vascular risk factors and subcortical vascular lesions, which reduce the degree of certainty of the diagnosis. Cortical atrophy is located in the right anterior temporal lobes (100% anteromedial, 100% anteroinferior, 70% anterolateral), left anterior temporal (90% anteromedial, 90% anteroinferior, 60% anterolateral), right prefrontal (30% basal, 50% dorsolateral, 20% medial) and left prefrontal (20% basal, 30% dorsolateral, 20% medial). CONCLUSIONS: Oral verbal production is influenced by level of education and, in a sample of patients with probable frontotemporal degeneration and logorrhoea, all the patients showed alterations in the anteroinferior and anteromedial regions of the right temporal lobe. PMID- 20013716 TI - [An atypical progression of rolandic epilepsy: the value of single-photon emission computerised tomography co-registered to magnetic resonance imaging]. AB - INTRODUCTION: The complicated forms of rolandic epilepsy progress with a continuous spike-wave pattern in slow-wave sleep. Experiments conducted in cats suggest that this pattern can only appear if there is bilateral thalamic insult. AIM. To determine whether thalamic hypoperfusion is associated with the complicated variants of rolandic epilepsy. PATIENTS AND METHODS: A group of 24 children were studied over a period of six years following their first epileptic seizure. During the follow-up an interictal magnetic resonance scan and single photon emission computerised tomography (SPECT) were performed. Results were examined to ascertain whether there were asymmetries in the distribution of cerebral blood flow through structures, using parametric statistical maps. The brain SPECT was performed when progression to atypical benign partial epilepsy in infancy was diagnosed and in typical forms of rolandic epilepsy when there was some mild neuropsychological deficit that led the specialist to suspect the existence of a focal cortical lesion. RESULTS: Bilateral thalamic hypoperfusion was found in all patients diagnosed with atypical benign partial epilepsy in infancy, which was correlated with the presence of continuous spike-waves during the slow-wave phase of non-REM sleep. CONCLUSIONS: Bilateral thalamic hypoperfusion seems to be a necessary condition for the atypical progression of rolandic epilepsy. PMID- 20013717 TI - [Reorganisation of the cerebral vasculature following ischaemia]. AB - INTRODUCTION AND DEVELOPMENT: Cerebral function needs a constant oxygen and glucose supply. This is why the regulation of cerebral blood flow is critical for the maintenance of neuronal function. Therefore, vascular system in adult brain is extremely stable and does not withstand big changes under physiological conditions. However, when blood flow is interrupted due to a focal cerebral ischemia the collateral tissue is partially affected, this is known as ischemic penumbra. Although its functionality is affected, this tissue is viable thanks to the collateral blood flow, and it releases angiogenic factors that induce proliferation of endothelial cells and migration of endothelial progenitor cells for the formation of new blood vessels. Angiogenesis induction and new vessel generation allow neurorepair processes, including neurogenesis and synaptogenesis. These two processes should be coupled with angiogenesis in order to contribute to functional recovery of patients who suffered a cerebral infarct. Therefore, angiogenesis could be one of the therapeutic options in ischemic stroke treatment. Nevertheless, some angiogenic factors such as vascular endothelial growth factor, platelet derived growth factor and angiopoyetin also increase vascular permeability which can produce hemorrhagic transformation. CONCLUSIONS: Hence, the knowledge of molecular mechanisms that regulate angiogenesis after an ischemic stroke could contribute to the development of a new therapeutic option based on angiogenesis as a vehicle to promote neurorepair and functional recovery. PMID- 20013718 TI - [The current state of the art concerning quality of life in Parkinson's disease: II. Determining and associated factors]. AB - INTRODUCTION: Health-related quality of life (HRQL) in Parkinson's disease (PD) is an area that is receiving a growing amount of interest as the new biopsychosocial model of medicine is adopted. AIM. This paper is the second part of a review of the current state of the art concerning HRQL in PD and focuses on its main determining and associated factors. DEVELOPMENT: A total of 56 studies are reviewed and the determining factors are grouped according to different variables, which may be disease-related (including motor and non-motor symptoms), sociodemographic (level of schooling, economic factors, gender and others), psychological and related to mental status (depression, anxiety, cognitive impairment, physician-patient relationship and others), and disability. The following characteristics of the studies were taken into account: the type of design (longitudinal or cross-sectional), the instrument used for measuring the HRQL (generic or specific), the statistical method employed (univariate or multivariate) and the sample size. CONCLUSIONS: Most of the studies are cross sectional and the majority use the 39-item Parkinson's Disease Questionnaire as a measure of HRQL with multivariate data analysis. The three most important factors determining HRQL in PD are depression, the stage of the disease and the time elapsed since onset of the disease. Nevertheless, the preferred methodology does not allow for causal inferences, due to the scarcity of longitudinal studies. PMID- 20013719 TI - [Nicotine as an antidepressant and regulator of sleep in subjects with depression]. AB - INTRODUCTION: Depression is a chronic and heterogeneous disorder manifesting itself by mood and sleep irregularities which ultimately can lead to suicide. Depressed individuals have a higher tendency to smoking compared to non-depressed people, and the interruption of this dependence produces even more depression. This situation may be reverted by the administration of anti-depressives and/or the use of nicotine replacement therapies. Taking into account these observations it will be explained how nicotine possibly acts in relation to two main neurotransmitters: serotonin and dopamine; and as a neuroprotective agent under a depressive condition. The effect of nicotine as a sleep regulator in depressed subjects will also be presented. AIM. To describe and discuss the evidences showing that nicotine has anti-depressive properties. DEVELOPMENT: In order of finding are presented in the following pages evidences showing that nicotine therapeutically can be used as an anti-depressive agent: general aspects of nicotine action (pharmacology, metabolism, nicotinic receptors), general features of depression, nicotine interactions in depressive disorders, and nicotine and the regulation of sleep. CONCLUSIONS: Even though the basis of the relationship 'nicotine-depression' is not clear, the suggested anti-depressive role of nicotine involves a direct action over dopaminergic and serotoninergic activity or a possible indirect action as a neuroprotective agent, events which may have therapeutic effects on the quality of sleep and enhancement of depressive symptoms. PMID- 20013720 TI - [Unusual cerebellar presentation of Wernicke's encephalopathy]. PMID- 20013721 TI - [Neuralgia of the intermediate nerve and facial palsy with or without herpes]. PMID- 20013722 TI - [Neonatal thrombosis of the cerebral venous sinuses]. PMID- 20013723 TI - [Energy-saving strategies in the treatment of fatigue in patients with multiple sclerosis]. PMID- 20013724 TI - History of Nephrology: a process confronted with changing times and of those who practiced it. PMID- 20013725 TI - Armenian medicine and diseases of the kidney. AB - The history of Armenian medicine is that of the common practice of medicine in caring for the sick through the ages, using mainly local medicinal herbs and natural products. Over the years, its practitioners persistently collected and recorded an expanding body of information on therapeutics and put it to use for the daily medical care of ordinary folks. Armenian medicine developed around the church and monasteries, which flourished during episodic periods of peace in an otherwise tumultuous and warring region. However, unlike the monastic medicine that developed in Europe, Armenian medicine maintained the rationality it had acquired from Greek medicine, and never resorted to magic, myth or amulets. Nor did it acquire or import saints. Armenian medicine is a classic example of the evolving art of therapeutics, whose record is preserved in extant manuscripts, saved over the centuries in monasteries, and now preserved in accessible collections. PMID- 20013726 TI - Isaac Judaeus Israeli: a Jewish founder of the origins of nephrology. AB - This paper discusses the Book of Urine in the Opera Omnia Isaci of Isaac Judaeus Israeli. Born in Egypt in the middle of the ninth century, Isaac Judaeus was considered a genius by his peers. The book accepted Galen's ideas and expanded them. Its original contribution was in the concept of the seeping of fluid and sediment from blood into the kidney and from the kidney to the bladder. This concept corresponds to glomerular filtration and tubular secretion. PMID- 20013727 TI - Kidney dysfunction, from the Arabic to the Byzantine world in 11th and 12th century southern Italy. AB - Greek classical medicine was not transmitted directly from classical antiquity to the Western Middle Ages by a continuous tradition, but passed through the Arabic world, where it had been preserved thanks to translations from Syriac and/or Greek into Arabic. From the Arabic world, Greek medicine arrived to the West, through the Greek- or the Latin-speaking communities in Sicily and the mainland, respectively. Remarkably enough, the scientific content of the text was scrupulously respected. PMID- 20013728 TI - Juxtaposition of Actuarius' versus Galen's ideas on renal physiology: the impact of 12 centuries. AB - Although some elements of renal physiology can be traced to scattered references in Greek medical writings, mainly in Hippocrates' and Erasistratus' works, it was Galen who made the first breakthrough observations regarding the function of the kidneys. He often wrote his observations not as a diatribe, but as a confrontation with other physicians, mainly of the Erasistratian School. He outlined the great importance of the disproportionally large blood supply of the kidneys, an over-proportion not observed in any other organ, rightly arguing that this is a teleological procedure to achieve satisfactory body clearance. He challenged the dominant idea that blood purification and hence the formation of urine took place in the large branches of the renal vessels. He strongly argued that the special structure of the kidney tissue was responsible for their cathartic ability. Although he had no idea of the histology of the kidney, he made some beautiful similitudes to explain the filtering capacity of the renal tissue. From Galen onwards, very few changes and additions can be traced in Greek and thereafter Byzantine medical texts, regarding renal physiology. It is Johannes Actuarius who, 12 centuries later, writes an extensive treatise on urine and presents his point of view on renal physiology, mainly as conclusions from the pathological findings of uroscopy. In the present paper, we compare the 2 physicians' approaches on the function of the kidneys, which can be attributed to the accumulation of medical knowledge over the 12 centuries that separate them. PMID- 20013729 TI - Practice, theory and authority in a Middle English medical text: "Barton's Urines Which He Treated at Tilney". AB - A medieval English text on the theory and practice of uroscopic analysis is found in British Library, Sloane MS 280, and Cambridge, St. John's College MS B.16. The treatise, the title of which is given in the fuller and earlier copy in Sloane 280 as "Barton's Urines Which He Treated at Tilney," offers an unusual mix of practical diagnostic methodology focused mainly on uroscopy, and a conceptual framework for that methodology which begins with humoral physiological theory but continues with digressions on astronomy, the calendar, Aristotelian psychology, reproductive anatomy and physiology, embryology and ensoulment. This paper discusses the possible authorship and dating of the original text, the author's intellectual interests and habits, his approach to his vernacular audience, and the relation of the treatise to the Latin authorities on which it draws, particularly but not exclusively Isaac Israeli and Giles of Corbeil. PMID- 20013730 TI - Ad vesice dolorem et ad eos qui urinam non faciunt (For bladder pain and when a person cannot urinate): nephrological disorders in Anglo-Saxon medicine. AB - The attitude of 19th century (and even of 20th century) scholars toward medieval and, particularly, Anglo-Saxon medicine has been of severe criticism. According to them it was filled with superstition and stupidities. However, in these last fifty years research has proved that, compared with the Continent, Anglo-Saxon England was not a backwater. At the end of the ninth century, medical compendia in Old English began to appear, similar in structure and contents to the Latin dynamidia and to the Latin herbals. These medical treatises were written in the vernacular of the Anglo-Saxons, not in Latin, the western European language for all significant and valuable works on medicine. Bladder, kidney and urinating problems are mentioned throughout the Old English medical treatises together with their cures, that is remedies from herbs and animals. These texts contain no theoretical reflections, only very concise descriptions of symptoms (pain in the bladder, in the kidneys, difficulty in urinating etc.), while prognosis is limited to affirmations such as "he will heal very quickly," "soon there will be no pain," "it will soon be healed," etc. Remedies are made basically out of a body of medicinal plants and materials which can be traced to Greek and Roman medicine. The remedies from plants reflect a wide rational and practical knowledge of medicinal herbs. As a matter of fact, not only there is no amuletic use of plants, but most of the herbs that appear in these recipes have diuretic or analgesic properties and have been in use for centuries. PMID- 20013731 TI - Reception of uroscopy in Scandinavia. AB - The purpose of this article is to look for possible traces of uroscopy in Scandinavia. The manuscripts and early editions on this subject limit the period of uroscopy in Scandinavia to 1450-1600, perhaps even beginning as late as 1557. In the first hundred years of this period, the most important texts were bought by students in foreign countries and taken home to their libraries, but it is doubtful that they were used. This impression is reinforced when we look at the first print in Danish of a medical text. Book printing began in Denmark in 1482, and this text by Christiern Pedersen is from 1533. It deals with herbs and thus continues a rich tradition for herbal medicine in Denmark, but uroscopy is not mentioned. The next editor of medical texts published 7 books in Danish from 1536 to 1557. The last one (1557) is on uroscopy, and in the preface the author, a practising physician, recommends knowledge of uroscopy as useful for his fellow citizens. His source is Lorenz Fries who originally did not want his edition in German to get into the hands of lay people - only less-learned colleagues. The Danish translator gives the impression that uroscopy is something entirely new in Denmark. In 1596 another medical textbook including uroscopy appeared. This is a translation of Ortolff von Bayrlandt's Arzneibuch by a Danish surgeon. It was intended for surgeons and was only reprinted twice. The medical books from the 17th century do not include traditional uroscopy. PMID- 20013732 TI - "De medicina et morbis" from De rerum naturis by Rabano Mauro. AB - De rerum naturis from the Monastery of Montecassino (MS codex 132) by Rabano Mauro, is a medieval encyclopedia issued around the year 1025, in the Abbey of Montecassino, during the period of Abbot Teobaldo. It is a copy of a more ancient text, written almost certainly in the Abbey of Fulda 2 centuries before, when Rabano Mauro was the abbot - that is, in the Carolingian age. In the Book 18 there is a chapter entitled "De medicina et morbis," in which we find related the fundamental principles on which studies of the human body, illness and the principal medicinal herbs were based. The text is not intended for teaching the medical art, but has the precise objective of helping to form the cultured Christian, trying to combine the old Greek-Roman tradition with a mystical vision, giving a moral application according to the dictates of Jewish-Christian religion. This text is indicative of the fusion of the scientific and religious worlds in the West during the Middle Ages. This interpretation of the universe will last for about a millennium, and only the Renaissance will be able to separate the 2 worlds again. PMID- 20013733 TI - Phosphorus: the philosopher's stone discovered in 1669. AB - Recently the importance in nephrology of phosphorus as phosphate has been highlighted by chronic renal failure patients, in whom the toxic effect of phosphate is widely acknowledged, given the association of phosphate serum level with cardiovascular risk. This association is not limited to chronic renal failure and hemodialysis patients as high serum phosphate. Recently high serum phosphate levels were associated with increased risk for cardiovascular disease in subjects free from chronic kidney disease, and cardiovascular disease as well, and with progression of atherosclerosis. It is useful to know the history of phosphorus from its discovery in 1669, because that history gives us more evidence to better understand the negative and/or toxic effects of high phosphate serum levels and to identify phosphorus as a physiologically crucial anion. PMID- 20013734 TI - Georg Philip Nenter and medicine "by notes" in the 18th century. AB - Even though it is true that the medical historiography of the 18th century is lacking in great scientific personalities, it is equally true that the entire century is characterized by continuous efforts to encapsulate the medical area of knowledge, acquired until then, in precise and systematic outlines, to serve the academic teaching of the subject of medicine. Georg Philip Nenter, a pupil of Georg Ernst Stahl and a professor of medicine in the University of Strasburg, could not help being influenced by that atmosphere. He added though, in our opinion, a touch of remarkable modernity. In fact his volume Fundamenta medicinae teoretico-pratica (Venice, 1735) is a wonderful collection of the notes taken during his lectures. The description of various diseases - renal diseases in particular - maintains a very systematic development of the subject through various chapters (definition, clinical manifestations, differential diagnosis, prognosis, treatment and examples of specific prescriptions), as if students were being addressed. PMID- 20013735 TI - Impotence in the 18th and 19th century: concepts of etiology and approaches to therapy. AB - The old word impotence is derived from the Latin word impotencia, which literally translated means "lack of power." Impotence, in the course of the history, has been attributed to mental pathology, anxiety, or demons or witches. Historically, the pharmacological treatments for impotence started in Greek times, when a myriad of herbal medications were applied locally to the genitals to enhance "sexual strength." In the 18th century, theories about the main factors inducing impotence saw it as an abnormal state of the fibers, a defect in the solid or liquid substances or a bad structure (tumor, inflammation, abscess, ulcer or foreign body). According to these mechanisms, when impotence depended on the state of the muscular fibers, treatment included a tepid bath and a clyster. In very fat or very weak people, who get particularly tired, it was important to use the remedies able to give energy to the fibers, such as ferrous mineral waters, for a month. Moreover, other suggestions were to ride a horse, to sleep few hours, to breathe good country air, to take a purge every 2 weeks, to drink half a glass of wine from Borgogne or to distract the mind continuously. In the 19th century, therapies regarding impotence included slight electric stimulation through the application of stimulators on the scrotum in the testis or epididymis areas, until pain was induced. In the same period, another method for treating impotence was flagellation. This method consisted of little flagellations with leather strips. PMID- 20013736 TI - Beginning of modern concept of inflammation: the work of Friedrich Daniel von Recklinghausen and Julius Friedrich Cohnheim. AB - In Rudolf Virchow's concept of inflammation, the basic alterations were derived from connective tissue cells, which underwent a marked metamorphosis. This cell based and static conception was fundamentally broadened and, in part, refuted in the ensuing decade by 2 of his scholars. Friedrich Daniel von Recklinghausen characterized the pus cells in acute inflammation and made the seminal observation of their contractility and mobility. He was the first who described the wandering leukocytes which were demonstrated in particular in experimental keratitis. He also showed that pus cells could migrate from the places of their origin in the interstitium to other tissues and epithelial cells. Von Recklinghausen in addition contributed to the concept of phagocytosis. The work of Julius Friedrich Cohnheim was focused on the mechanisms involved in the extravasation of leukocytes from the blood vessels in the inflamed mesentery of the frog and carefully described the time-dependent alterations: dilatation of the arteries and veins, adhesion of colorless cells to the endothelial cells, and the subsequent transmigration from the capillaries and venules into the interstitial space. In the last few decades, experimental and clinical studies using modern techniques have fully confirmed and extended these basic observations made by von Recklinghausen and Cohnheim more than 100 years ago. PMID- 20013737 TI - Tadeusz Reichstein: from description of coffee aroma to discoveries of cortisone and aldosterone. AB - Tadeusz Reichstein was born in Wloclawek (Poland) into a Polish-Jewish family. His family emigrated to Switzerland, and he was educated in the Technical University of Zurich, becoming an engineer of chemistry. Thus he started his scientific career, firstly in Zurich and later in Basel. In his very busy life, he developed a method of vitamin C synthesis enabling industrial production of this important compound. Later, Reichstein isolated and synthesized such important adrenocortical hormones as cortisone, desoxycorticosterone and aldosterone. He received several awards but the most important was a Nobel Prize in Physiology or Medicine (1950). Reichstein spent the last years of his life in his own botanical garden and laboratory working on fern cytogenetics and the relations between different species of this archaic plant. Despite extraordinary and still valuable scientific achievements, he was always a very modest man with a humanistic attitude. PMID- 20013738 TI - Water, from Gilgamesh Epic to Nobel Laureate Richard Feynman: a look into polywater and the memory of water. AB - Water is a complex source of imagination, dreams and rituals, where cultural differences ebb and flow, where a plethora of meanings and interpretations interlink and wash over one another. Water has an ambivalent character as stated in most of the ancient cosmogonies and in the Epic of Gilgamesh. Water's composition was discovered by the London scientist Henry Cavendish in about 1781. Although it is an apparently simple molecule (H2O), it has a highly complex and anomalous character. The anomalous properties of water are those where the behavior of liquid water is quite different from what is found with other liquids. As often stated, life depends indeed on these anomalous properties of water. Notably there are 12 phase, 22 density, 12 material, 11 thermodynamic and 9 physical anomalies. A powerful look into the water molecule was given by Nobel Prize recipient Richard P. Feynman as published in Six easy pieces. A look into the most recent quest for more knowledge about water leads us to the concept of pathological science. The cases of "polywater" and "the memory of water" are indeed paradigmatic episodes of fraudulent research published in journals with high impact factors. In conclusion, men came out of water engineered to handle water, and water greatly affects mythology and philosophy and is a strong presence in the arts and science. PMID- 20013739 TI - The nature of water: Greek thought from Homer to Acusilaos. AB - Greek philosophy finds its roots in the myth of Homer's and Hesiod's poems and especially in Orphism which introduced the concept of a soul separated from the body with an independent principle, psiche (soul), to be rewarded or punished after death. Orphism was an important step in Greek culture. It introduced the divine into man, the soul which does not die with the body and reincarnates. From Orphism started the need of rituals capable of separating the spirit from the body. From Homer to Acusilaos, water was a very important element which connected humans and gods, long before Thales of Miletus defined it the arche. PMID- 20013740 TI - The nature of water: Thales' arkhe. AB - Thales was born into a noble family of Phoenician origin at the time of the 25th Olympiad (floruit 585 bc; he was 40 in the year of the solar eclipse. He had no teachers but had occasion to learn from Egyptian priests. He developed into a scholar and politician very much appreciated by Heraclitus, Herodotus and Democritus, and was always considered a man of practical wisdom. He was probably the first to speak about the immortality of the soul. He is listed as the first of many unmarried men who paved the road for philosophy. For Diogenes Laertius (Lives and Opinions of Eminent Philosophers), he was the instructor of Anaximander. Thales, the man who first discovered how to draw a right-angle triangle in a circle, was the first philosopher of nature (physis). "Philosophy begins with Thales," pointed out Bertrand Russell in 1961. This honor had been conceded also by Aristotle: "Anaximander, Thales' pupil, founded the Ionian tradition of philosophy." Many explanations may be given for the importance of water, including its importance for living processes, the economic role of the Nile, the importance of the port for Miletus and the fact that Ocean and Thetys were in Homer's tradition progenitors of the world. PMID- 20013741 TI - The nature of water: excerpts from Pythagoras, Xenophanes, Heraclitus and Parmenides. AB - Water was a prominent substance with Pythagoras, Xenophanes, Heraclitus and Parmenides, who flourished in the years 530-490 bc. The basic Pythagorean elements were earth and fire, and between them there were 2 intermediate entities (water and air), which were instrumental and indispensable components of specific solids. All things are a blend of different elements. For Xenophanes, "All things that come into being and grow are earth and water," "We all originated from earth and water" and "And in certain caves water drips down."For Heraclitus water is an ambivalent substance: "One cannot bathe in the same river on two occasions." "The sea is the safest and the most polluted water, for fish it is healthy and gives life, for men it is unhealthy and causes death." "Fire experiences the death of earth, air experiences that of fire, water experiences the death of air and the earth that of water." Parmenides was a man who sought the truth through reasoning and was, according to Hegel, the founder of Western philosophy. He built a dualist theory of the cosmos based on heat and cold, fire and earth - the former as a cause, the latter as substrate. The former unified, the latter separated. According to Aristotle, Parmenides considered air and water as mixtures of earth and fire. PMID- 20013742 TI - Historical perspective on the role of the kidney in acid-base regulation. AB - Early observations on the acidity of normal urine by J. B. von Helmont (1527 1644) and on urine content of sulfate, phosphate and carbonate by J. J. Berzelius (1779-1848), followed by the studies of Bence Jones (1813-1878) on the connection between food, nutrition and urine acidity, pointed to the role of the kidney in regulation of acid-base status in humans and animals. The next important steps in this field of science were studies by F. Walter (1877) on decreased "alkali" in blood and increased ammonia in the urine of dogs after infusion into their blood of hydrochloric acid, and the observations of B. Naunyn (1939-1925) and O. Minkowski (1853-1931) on the presence of beta-hydroxybutyric acid in urine and on increased ammonia excretion in urine from patients with diabetic coma. Also it was found that patients with uremia had decreased titratable "alkali' in blood (R. von Jaksch 1855-1947) and reduced ability to excrete ammonia (W. W. Palmer and L. J. Henderson 1915). Finally, studies by R. F. Pitts (1908-1977) defined the role of the kidney in reabsorption of bicarbonate in the tubules and linked hydrogen secretion to sodium excretion in the urine. PMID- 20013743 TI - Thomas Addis, 1881-1949, clinical scientist, hematologist and pioneering nephrologist: a brief biography. AB - Thomas Addis is an important figure in the history of nephrology. Born in Scotland and trained in Edinburgh, he came to San Francisco in 1911 to the new Stanford School of Medicine to run the clinical laboratory. Over the next 38 years, he made many contributions to renal physiology, the investigation of the structure and function of the kidneys in Bright's disease, and studies of kidney growth, hypertrophy and protein metabolism. Largely forgotten today, he was one of the first to use urea clearance as a measure of kidney function and was the first to systematize examination of the urinary sediment - the Addis count. He was also a leader in the use of diet and rest in the treatment of Bright's disease. Unknown to most nephrologists, before he came to the United States during the first 6 years following his graduation, he became one of the leading clinical investigators in hematology. His special interests were the mechanism of blood clotting and hemophilia. He was the first to transfuse fresh blood into a hemophiliac patient and show that this shortened the patient's clotting time. Addis was a great, if eccentric, personality as well as a great scientist. He was beloved by his many colleagues and students and was honored in his lifetime both in the United States and Great Britain. PMID- 20013744 TI - Blackwater fever: the rise and fall of an exotic disease. AB - After several descriptions by Hippocrates and a single possible medieval description by Gilles de Corbeil, a severe febrile illness accompanied by the passage of dark urine burst upon the medical scene in West Africa in 1819, described by an English surgeon named Tidlie. Most of his patients died within a few days. Further reports appeared from tropical regions until the turn of the century, J. Farrell Easmon having given the condition the name blackwater fever in 1884. Controversy raged about its relationship to malaria, as well as over its treatment with cinchona bark and quinine. Evidence evolved that it was a complication of falciparum malaria in which hemoglobinuria causing acute renal failure resulted from massive quinine-induced lysis of red blood cells. People with red cell abnormalities such as glucose-6-phosphate dehydrogenase deficiency proved particularly prone to developing it. Its incidence fell as more mildly acting antimalarial drugs replaced quinine. Several enigmatic issues bedeviled understanding of it, but a careful analysis of its historical development has enabled resolution of each of these. PMID- 20013745 TI - Francophone clinical nephrology from 1945 to 1960. AB - After the Second World War, clinical nephrology in Francophone countries had 4 leaders: in Paris, P. Vallery Radot with his assistants P. Milliez and J. Hamburger and their own groups; and also in Paris, M. Derot with J. J. Bernier and M. Legrain; in Bruxelles, P. Govaerts with P. P. Lambert; and in Geneve, R. Mach. To these one must add J. Traeger of Lyon and J. M. Suc of Toulouse who later joined the group. It all started in 1947-1948 around the Societe de Pathologie Renale. In the current paper, the author describes the main 4 fields of development: water and electrolyte balance, acute anuria and dialysis, renal biopsy and classification of nephritides and finally transplantation under the direction of J. Hamburger and R. Kuss both closely linked with J. Dausset and G. Mathe. These contributions had a broad impact on the history of modern clinical nephrology, which was acknowledged and crowned by the organization of the first Congres International de Nephrologie by Hamburger and Mach, the development by which the specialty was acknowledged and the International Society of Nephrology (ISN) created with its first journal, Nephron. Hamburger, the first president of the ISN, had many difficulties to overcome to carry this goal through. PMID- 20013746 TI - Low salt diet and treatment of hypertension: an old story. AB - A relatively low salt intake is nowadays considered one of the characteristics of a healthy diet in the Western world because several disorders appear to be unfavorably affected by excessive salt intake with the diet. The first notion about a relation between salt intake and blood pressure traces back to 2500 bc in an ancient Chinese medical textbook. This paper focuses on studies about salt and hypertension in the first half of the 20th century. The first papers in this field were published from the beginning of the century, but due to a modest scientific content were still not considered in the 1940s to provide sufficient evidence in favor of a salt restriction in hypertensive patients. A major practical contribution came from the Kempner rice diet, an effective antihypertensive dietary treatment which included a severe restriction of salt intake. After that, several studies in animals and humans showed that, with regard to the antihypertensive effect, the key element of the Kempner diet was the low salt content. By the first years of the 1950s, the evidence was already available that salt restriction is an effective antihypertensive treatment and that adherence to the treatment should be assessed by monitoring urinary electrolytes. PMID- 20013747 TI - Diet and the kidney. AB - This is a short synopsis of the importance of nutrition and disease, which is especially centered on the achievements concerning renal diseases. With regard to renal nutrition, the paper analyzes the contributions of Beale, Peters and van Slyke, Addis, and Borst, and discusses the advent of the Giordano-Giovannetti diet and its modifications. PMID- 20013748 TI - Nephrology and Doctors Honoris Causa at P. J. Safarik University in Kosice in the years 1993-2006 (Slovak Republic). AB - The authors present a brief history of the activities of 7 important and well known foreign nephrologists in Kosice and in the region of Eastern Slovakia who were awarded the honorary title Doctor Honoris Causa by P. J. Safarik University in Kosice. The above-mentioned professors presented their papers as guest professors to the students of the Medical Faculty of P. J. Safarik University and in meetings of medical societies and at many symposia and congresses with international participation in the region of Eastern Slovakia. All of the awarded nephrologists have visited the Faculty Hospital of L. Pasteur, the Fourth Internal Clinic and the Nephrological Clinic. During their stays, they stimulated the thinking of researchers in other metabolic studies within clinical nephrology and toxicology. In addition they contributed to the establishment in 1997 of the Nephrological Clinic of the Medical Faculty of P. J. Safarik University and the Faculty Hospital of L. Pasteur, the first one of its kind in the Slovak Republic. PMID- 20013749 TI - The Mediterranean diet: a matter of history, tradition, culture and health. AB - We can state that unhealthy diets are among the top 10 causes of mortality in Western countries. The Mediterranean diet is gaining popularity because it encourages weight loss, improving the quality of life by offering a variety of healthy and palatable foods. Moreover, it ensures adequate intake of fruits, vegetables, nuts, fish, fibers, legumes, cereals and olive oil, which have been associated with a longer lifespan of people thanks to their anti-inflammatory and antioxidant properties. Thus the Mediterranean diet could act as therapy in inflammatory diseases including cardiovascular disease, obesity, type 2 diabetes, metabolic syndrome and chronic renal failure. PMID- 20013750 TI - Klaus Hierholzer (1929-2007) and his impact on our understanding of the renal effects of steroid hormones. AB - Klaus Hierholzer (1929-2007) dissected various functions influenced by steroids in the distal tubule and showed that aldosterone in low doses reversed the sodium and potassium transport defect in adrenalectomized rats, through a rapid activation of Na+,K+-ATPase. Subsequent studies addressed the role of 11-beta hydroxysteroid oxidoreductase (11-HSD) and showed that the undisturbed functioning of 11-HSD is a prerequisite for selective mineralocorticosteroid regulation of epithelial transport. Another set of original experiments showed that 11-HSD was equally important in the distal colon, thus establishing that the large intestine acts in parallel with the distal nephron. Hierholzer, born in Konstanz on June 8, 1929, was laureated in medicine on May 25, 1954. Subsequently he worked at the Department of Pharmacology of the University of Freiburg, Cornell University with J. F. Pitts, the Department of Medicine of the University of Frankfurt-am-Main, the University of Copenhagen with H. H. Ussing, and the Institute of Physiology of the Freie Universitaet in Berlin where he became full professor and head of the Institute of Clinical Physiology in 1968. He held that position until 1998. He died in Allensbach in the family house on February 27, 2007. Hierholzer was a member of the Naturforscher Leopoldina Academy and of many other scientific societies, including the Academy of Science and Technology in Berlin, and received various awards including an honorary professorship at the University of Naples, the Bezold Medal, the Volhard Medal, the Schoeller/Junkman Award, and the Malpighi Medal (in memoriam). He published nearly 300 papers including various seminal books. Noteworthy also are his papers on the history of physiology of the kidney and acid-base balance. A total of 26 scientists who trained in his laboratory became professors. PMID- 20013751 TI - Interactions of aluminum nanoparticles with human epidermal keratinocytes. AB - Aluminum nanoparticles (Al NP) have been used in applications as diverse as drug delivery, material surface coatings and an ingredient for solid rocket fuel in military explosives and artillery. Although Al NP are used in many civilian and military applications, the health and safety implications of these nanosize particles are not known. To understand the interactions and biological activity of Al NP in human cells, cultured human neonatal epidermal keratinocytes (HEK) were exposed for 24 h to 50 and 80 nm Al NP in concentrations from 4.0 to 0.0004 mg ml(-1) to assess the cytotoxicity and inflammatory potential. UV-Vis measurements and nanoparticle controls revealed that the Al NP interact with the assay dyes. Viability did not decrease in HEK exposed to both the 50 and the 80 nm Al NP at all treatment concentrations with MTT, CellTiter 96 AQueous One (96 AQ) and alamar Blue (aB) viability assays. The 96 AQ and aB assays interact with the Al NP less than MTT, and proved to be the best assays to use with these Al NP. TEM depicted Al NP localized within the cytoplasmic vacuoles of the cells. Cytokine data was variable, indicating possible nanoparticle interactions with the cytokine assays. These studies illustrate the difficulties involved in assessing the biological safety of nanomaterials such as Al NP due to media- and temperature-dependent particle agglomeration and nanoparticle interactions with biomarkers of cytotoxicity. PMID- 20013752 TI - Generation of micropatterned substrates using micro photopatterning. AB - Micro photopatterning (microPP) has been developed to rapidly test and generate different patterns for extracellular matrix adsorption without being hindered by the process of making physical stamps through nanolithography techniques. It uses two-photon excitation guided through a point-scanning confocal microscope to locally photoablate poly(vinyl) alcohol (PVA) thin films in user-defined computer controlled patterns. PVA thin films are ideal for surface blocking, being hydrophilic substrates that deter protein adsorption and cell attachment. Because gold substrates are not used during microPP, all live-cell fluorescent-imaging techniques including total internal reflection fluorescence microscopy of GFP linked proteins can be performed with minimal loss of fluorescence signal. Furthermore, because microPP does not require physical stamps for pattern generation, multiple ECMs or other proteins can be localized within microns of each other. This unit details the setup of microPP. It also provides troubleshooting techniques. PMID- 20013753 TI - In vivo imaging of signal transduction cascades with probes based on Forster Resonance Energy Transfer (FRET). AB - Genetically encoded FRET probes enable us to visualize a variety of signaling events such as protein phosphorylation and G-protein activation in living cells. This unit focuses on FRET probes wherein both the donor and acceptor are fluorescence proteins and incorporated into a single molecule, i.e., a unimolecular probe. Advantages of these probes lie in their easy loading into cells, simple acquisition of FRET images, and clear evaluation of data. We have developed FRET probes for Ras-superfamily GTPases, designated Ras and interacting protein chimeric unit (Raichu) probes. We hereby describe strategies to develop Raichu-type FRET probes, procedures for their characterization, and acquisition and processing of images. Although improvements upon FRET probes are still based on trial-and-error, we provide practical tips for their optimization and briefly discuss the theory and applications of unimolecular FRET probes. PMID- 20013754 TI - Interference reflection microscopy. AB - Interference reflection microscopy (IRM) is an optical technique used to study cell adhesion or cell mobility on a glass coverslip. The interference of reflected light waves generates images with high contrast and definition. IRM can be used to examine almost any cell that will rest upon a glass surface, although it is most useful in examining sites of close contact between a cell and substratum. This unit presents methods for obtaining IRM images of cells with particular emphasis on IRM imaging with a laser scanning confocal microscope (LSCM), as most LSCM are already capable of recording these images without any modification of the instrument. Techniques are presented for imaging fixed and live cells, as well as simultaneous multi-channel capture of fluorescence and reflection images. PMID- 20013755 TI - Fluorescence correlation spectroscopy in living cells: a practical approach. AB - Fluorescence correlation spectroscopy (FCS) is a single-molecule fluorescence technique used to monitor molecular dynamics. It can be applied to living cells expressing fluorescently labeled proteins and lipids to determine the diffusion timescales and the total number of diffusing fluorescent molecules in the cell. A practical step-by-step approach to performing FCS with a commercial spectroscopy/microscopy system, the Zeiss Confocor 3, how to set up live-cell FCS experiments, acquire reliable data, and finally how to analyze the data acquired, are all described in this unit. PMID- 20013756 TI - Treatment of post-traumatic stress disorder in children using cognitive behavioural writing therapy. AB - OBJECTIVES: This study evaluated the effectiveness of Cognitive Behavioral Writing Therapy (CBWT) in 23 children (age 8-18 years) in the Netherlands, who experienced a range of single and recurrent traumatic experiences. CBWT uses exposure, cognitive restructuring and social sharing. METHODS: At pre-test, post test and follow-up, post-traumatic stress disorder (PTSD) symptoms, depressive symptoms, trauma-related cognitions and general behavioural problems were assessed. RESULTS: At post-test there was a significant reduction of all symptoms, and this effect was maintained at 6 months follow-up. The mean amount of treatment sessions needed was 5.5. CONCLUSIONS: This study shows that short term CBWT is a potentially effective intervention for clinically referred traumatized children. There is now a clear need of establishing the effectiveness of CBWT in a randomized, controlled trial. PRACTICE IMPLICATION: This first study indicates CBWT is a promising treatment, which can easily be used in clinical practice. PMID- 20013757 TI - Exploring therapeutic alliance in brief inpatient psychotherapy: a preliminary study. AB - BACKGROUND: Therapeutic alliance is one of the most widely investigated variables in psychotherapy research but few studies have explored its role in inpatient psychotherapy. Many factors likely contribute to the lack of inpatient alliance research including the short length of hospital stays, complexity of patient psychopathology and the burdensome quality of most alliance scales. This paper reports on the development and initial application of two comparable brief scales designed to capture patient and therapist alliance ratings. METHOD: Participants were 20 patients receiving supportive-expressive inpatient psychotherapy. The patients were predominantly depressed women. Baseline measures of distress, symptom severity and functioning were obtained at the first and third sessions. Measures of alliance were obtained at the second session. RESULTS: The brief alliance scales demonstrated adequate internal consistency and the individual items had good adjusted item-to-scale correlations. Consistent with the broad alliance literature, we found that patients rated alliance higher than therapists, patient and therapist alliance ratings were not significantly correlated, and level of functioning was significantly associated with both patients and therapists' alliance ratings. The perceived depth of psychotherapy was also significantly associated with alliance. Unexpectedly, alliance ratings were also negatively associated with improvement during hospitalization. CONCLUSIONS: Overall, the study demonstrates both the feasibility and potential benefit of conducting inpatient psychotherapy research. PMID- 20013758 TI - Analysis of phenolic compounds in health care products by low-pressure liquid chromatography with monolithic column and chemiluminescent detection. AB - This paper presents a new application for monolithic columns with low-pressure chromatographic separation using an flow injection analysis configuration with chemiluminescent detection for the determination of a mixture of phenolic compounds: phloroglucinol, 2,4-dihydroxybenzoic acid, salicylic acid, methyl paraben and n-propyl gallate. The procedure consists of the separation of these compounds on a reverse-phase ultra-short monolithic column with pH 3.0 acetate buffer and 5% acetonitrile as carrier phase. The detection is based on a chemiluminescence measurement coming from Ce(IV)-Rhodamine 6G chemistry with the incorporation of two different chemiluminescent chemical conditions in the chromatographic setup in order to enhance the sensitivity for the different phenolic compounds. All separation and detection variables were optimized to propose a determination method. The analysis is performed in 280?s, with the sampling frequency being some 13 h(-1) . The calibration function is a double reciprocal function obtaining good results within two orders of magnitude. The limits of detection were 8.8 * 10 (-8) m (phloroglucinol), 2.7 * 10 (-8) m (2,4 dihydroxybenzoic acid); 2.3 * 10 (-8) m (salicylic acid); 5.2 * 10 (-8) m (methyl paraben) and 4.1 * 10 (-6) m (n-propyl gallate), and the relative standard deviations at a medium level of the linear range were 4.4% (phloroglucinol), 2.8% (2,4-dihydroxybenzoic acid), 5.2% (salicylic acid), 3.6% (methyl paraben) and 6.8% (n-propyl gallate). The method was applied and validated satisfactorily for the determination of these compounds in healthcare products, comparing the results against an HPLC reference method. PMID- 20013759 TI - A dissolved oxygen sensor based on composite fluorinated xerogel doped with platinum porphyrin dye. AB - A new functional fluorinated material taking n-propyltrimethoxysilicane (n-propyl TriMOS) and 3,3,3-trifluoropropyltrimethoxysilicane (TFP-TriMOS) as precursors was applied to construct a novel dissolved oxygen sensing film. The sensing film was fabricated by dip-coating the functional fluorinated material-doped [meso tetrakis(pentafluorophenyl) porphyinato] platinum(II) (PtF(20) TPP) onto a glass slide. The oxygen sensing film exhibited a good linear relationship, fast response time, long stability and high sensitivity to dissolved oxygen. In the developed optical oxygen sensor, an LED and a photodiode were composed to construct a back-detection optical system not needing an optical fiber based on fluorescence intensity detection. The smart optical oxygen sensor based on the PtF(20) TPP fluorescence quenching possesses the advantages of portability and low cost and can be applied to the dissolved oxygen in situ monitoring in seawater. PMID- 20013760 TI - Working Alliance Inventory-Short Revised (WAI-SR): psychometric properties in outpatients and inpatients. AB - The Working Alliance Inventory-Short Revised (WAI-SR) is a recently refined measure of the therapeutic alliance that assesses three key aspects of the therapeutic alliance: (a) agreement on the tasks of therapy, (b) agreement on the goals of therapy and (c) development of an affective bond. The WAI-SR demonstrated good psychometric properties in an initial validation in psychotherapy outpatients in the USA. The generalizability of these findings is limited because in some countries a substantial portion of individual psychotherapy is delivered in inpatient settings. This study investigated and compared the psychometric properties of the WAI-SR in German outpatients (N = 88) and inpatients (N = 243). In both samples reliability (alpha > 0.80) and convergent validity with the Helping Alliance Questionnaire were good (r > 0.64). Confirmatory factor analysis showed acceptable to good model fit for the proposed Bond-Task-Goal model in both samples. Multi-group analysis demonstrated that the same constructs were measured across settings. Alliance ratings of outpatients and inpatients differed regarding the overlap of alliance components and the magnitude of the alliance ratings: The differentiation of the alliance components was poorer in inpatients and they reported lower alliances. Unique aspects of the alliance in inpatient treatment are discussed and a need for further research on the alliance in inpatient settings is pointed out. Overall, the WAI-SR can be recommended for alliance assessment in both settings. PMID- 20013761 TI - Attachment and symptoms of psychopathology: early maladaptive schemas as a cognitive link? AB - This study investigated whether early maladaptive schemas can explain the relation between attachment anxiety and avoidance dimensions and symptoms of psychopathology. For this purpose, 289 Flemish, Dutch-speaking, late adolescents participated on a questionnaire study. Using a non-parametric re-sampling approach, we investigated whether the association between attachment and psychopathology was mediated by early maladaptive schemas. Results indicate that the association between attachment anxiety and psychopathology is fully mediated by cognitions regarding rejection and disconnection and other-directedness. The association between attachment avoidance and psychopathology is partly mediated by cognitions regarding rejection and disconnection. PMID- 20013763 TI - N-heterocyclic carbene (NHC)-stabilized silanechalcogenones: NHC-->Si(R2)=E (E=O, S, Se, Te). AB - A series of N-heterocyclic carbene-stabilized silanechalcogenones 2 a,b (Si=O), 3 a,b (Si=S), 4 a,b (Si=Se), and 5 a,b (Si=Te) are described. The silanone complexes 2 a,b were prepared by facile oxygenation of the carbene-silylene adducts 1 a,b with N(2)O, whereas their heavier congeners were synthesized by gentle chalcogenation of 1 a,b with equimolar amounts of elemental sulfur, selenium, and tellurium, respectively. These novel compounds have been isolated in a crystalline form in high yields and have been fully characterized by a variety of techniques including IR spectroscopy, ESIMS, and multinuclear NMR spectroscopy. The structures of 2 b, 3 a, 4 a, 4 b, and 5 b have been confirmed by single-crystal X-ray crystallography. Due to the NHC-->Si donor-acceptor electronic interaction, the Si=E (E=O, S, Se, Te) moieties within these compounds are well stabilized and thus the compounds possess several ylide-like resonance structures. Nevertheless, these species also exhibit considerable Si=E double bond character, presumably through a nonclassical Si=E pi-bonding interaction between the chalcogen lone-pair electrons and two antibonding Si-N sigma* orbitals, as evidenced by their high stretching vibration modes and the shortening of the Si-E distances (between 5.4 and 6.3%) compared with the corresponding Si-E single-bond lengths. PMID- 20013762 TI - Secondary interactions or ligand scrambling? Subtle steric effects govern the iridium(I) coordination chemistry of phosphoramidite ligands. AB - The like and unlike isomers of phosphoramidite (P*) ligands are found to react differently with iridium(I), which is a key to explaining the apparently inconsistent results obtained by us and other research groups in a variety of catalytic reactions. Thus, the unlike diastereoisomer (aR,S,S)-[IrCl(cod)(1 a)] (2 a; cod=1,5-cyclooctadiene, 1 a=(aR,S,S)-(1,1'-binaphthalene)-2,2'-diyl bis(1 phenylethyl)phosphoramidite) forms, upon chloride abstraction, the monosubstituted complex (aR,S,S)-[Ir(cod)(1,2-eta-1 a,kappaP)](+) (3 a), which contains a chelating P* ligand that features an eta(2) interaction between a dangling phenyl group and iridium. Under analogous conditions, the like analogue (aR,R,R)-1 a' gives the disubstituted species (aR,R,R)-[Ir(cod)(1 a',kappaP)(2)](+) (4 a') with monodentate P* ligands. The structure of 3 a was assessed by a combination of X-ray and NMR spectroscopic studies, which indicate that it is the configuration of the binaphthol moiety (and not that of the dangling benzyl N groups) that determines the configuration of the complex. The effect of the relative configuration of the P* ligand on its iridium(I) coordination chemistry is discussed in the context of our preliminary catalytic results and of apparently random results obtained by other groups in the iridium(I)-catalyzed asymmetric allylic alkylation of allylic acetates and in rhodium(I)-catalyzed asymmetric cycloaddition reactions. Further studies with the unlike ligand (aS,R,R)-(1,1'-binaphthalene)-2,2'-diyl bis{[1-(1-naphthalene-1 yl)ethyl]phosphoramidite} (1 b) showed a yet different coordination mode, that is, the eta(4)-arene-metal interaction in (aS,R,R)-[Ir(cod)(1,2,3,4-eta-1 b,kappaP)](+) (3 b). PMID- 20013764 TI - Spatially controllable DNA condensation by a water-soluble supramolecular hybrid of single-walled carbon nanotubes and beta-cyclodextrin-tethered ruthenium complexes. AB - A supramolecular hybrid is prepared by the supramolecular surface modification of single-walled carbon nanotube (SWCNT) with cationic beta-cyclodextrin-tethered ruthenium complexes through a spacer molecule that contains both an adamantane and a pyrene moiety. By employing the supramolecular hybrid, spatially controllable DNA condensation along the SWCNT skeleton is achieved by anchoring cationic ruthenium complexes on the surface. Furthermore, because of the unique physiological properties of SWCNTs, the cationic supramolecular hybrid can be used as a nonviral gene delivery system with the ruthenium complexes as a fluorescent probe to monitor uptake of DNA by cells. PMID- 20013765 TI - Asymmetric synthesis of alpha-tocopherol. AB - Alpha-tocopherol was synthesized from a chiral intermediate alpha-hydroxy ester by means of two ring-closing methods to yield the chromanol in 94% diastereomeric excess. PMID- 20013766 TI - Catalytic double stereoinduction in asymmetric allylic alkylation of oxindoles. AB - A highly regio-, diastereo-, and enantioselective allylic alkylation reaction of 3-monosubstituted oxindoles catalyzed by molybdenum is described. The reaction is affected by the electronic and steric variations of the nucleophile. The use of appropriate N-protecting group is particularly important for achieving high regio and diastereoselectivity. Products from this reaction, containing vicinal quaternary-tertiary stereogenic centers, are valuable synthetic intermediates and should find utility in alkaloid synthesis. PMID- 20013767 TI - Asymmetric hydrogenation with water/silane as the hydrogen source. PMID- 20013768 TI - Mechanistic investigation of the dipolar [2+2] cycloaddition-cycloreversion reaction between 4-(N,N-dimethylamino)phenylacetylene and arylated 1,1 dicyanovinyl derivatives to form intramolecular charge-transfer chromophores. AB - The kinetics and mechanism of the formal [2+2] cycloaddition-cycloreversion reaction between 4-(N,N-dimethylamino)phenylacetylene (1) and para-substituted benzylidenemalononitriles 2 b-2 l to form 2-donor-substituted 1,1-dicyanobuta-1,3 dienes 3 b-3 l via the postulated dicyanocyclobutene intermediates 4 b-4 l have been studied experimentally by the method of initial rates and computationally at the unrestricted B3LYP/6-31G(d) level. The transformations were found to follow bimolecular, second-order kinetics, with DeltaH(exp)(not equal)=13-18 kcal mol-1, DeltaS(exp)(not equal) approximately -30 cal K-1 mol-1, and DeltaG(exp)(not equal)=22-27 kcal mol-1. These experimental activation parameters for the rate determining cycloaddition step are close to the computational values. The rate constants show a good linear free energy relationship (rho=2.0) with the electronic character of the para-substituents on the benzylidene moiety in dimethylformamide (DMF), which is indicative of a dipolar mechanism. Analysis of the computed structures and their corresponding solvation energies in acetonitrile suggests that the rate-determining attack of the nucleophilic, terminal alkyne carbon onto the dicyanovinyl electrophile generates a transient zwitterion intermediate with the negative charge developing as a stabilized malononitrile carbanion. The computational analysis predicted that the cycloreversion of the postulated dicyanocyclobutene intermediate would become rate-determining for 1,1-dicyanoethene (2 m) as the electrophile. The dicyanocyclobutene 4 m could indeed be isolated as the key intermediate from the reaction between alkyne 1 and 2 m and characterized by X-ray analysis. Facile first-order cycloreversion occurred upon further heating, yielding as the sole product the 1,1-dicyanobuta-1,3-diene 3 m. PMID- 20013770 TI - Photochemical reduction of carbon dioxide catalyzed by a ruthenium-substituted polyoxometalate. AB - A polyoxometalate of the Keggin structure substituted with Ru(III), (6)Q(5)[Ru(III)(H(2)O)SiW(11)O(39)] in which (6)Q=(C(6)H(13))(4)N(+), catalyzed the photoreduction of CO(2) to CO with tertiary amines, preferentially Et(3)N, as reducing agents. A study of the coordination of CO(2) to (6)Q(5)[Ru(III)(H(2)O)SiW(11)O(39)] showed that 1) upon addition of CO(2) the UV/Vis spectrum changed, 2) a rhombic signal was obtained in the EPR spectrum (g(x)=2.146, g(y)=2.100, and g(z)=1.935), and 3) the (13)C NMR spectrum had a broadened peak of bound CO(2) at 105.78 ppm (Delta(1/2)=122 Hz). It was concluded that CO(2) coordinates to the Ru(III) active site in both the presence and absence of Et(3)N to yield (6)Q(5)[Ru(III)(CO(2))SiW(11)O(39)]. Electrochemical measurements showed the reduction of Ru(III) to Ru(II) in (6)Q(5)[Ru(III)(CO(2))SiW(11)O(39)] at -0.31 V versus SCE, but no such reduction was observed for (6)Q(5)[Ru(III)(H(2)O)SiW(11)O(39)]. DFT-calculated geometries optimized at the M06/PC1//PBE/AUG-PC1//PBE/PC1-DF level of theory showed that CO(2) is preferably coordinated in a side-on manner to Ru(III) in the polyoxometalate through formation of a Ru-O bond, further stabilized by the interaction of the electrophilic carbon atom of CO(2) to an oxygen atom of the polyoxometalate. The end-on CO(2) bonding to Ru(III) is energetically less favorable but CO(2) is considerably bent, thus favoring nucleophilic attack at the carbon atom and thereby stabilizing the carbon sp(2) hybridization state. Formation of a O(2)C-NMe(3) zwitterion, in turn, causes bending of CO(2) and enhances the carbon sp(2) hybridization. The synergetic effect of these two interactions stabilizes both Ru-O and C-N interactions and probably determines the promotional effect of an amine on the activation of CO(2) by [Ru(III)(H(2)O)SiW(11)O(39)](5-). Electronic structure analysis showed that the polyoxometalate takes part in the activation of both CO(2) and Et(3)N. A mechanistic pathway for photoreduction of CO(2) is suggested based on the experimental and computed results. PMID- 20013769 TI - Solvent-resistant nanofiltration for product purification and catalyst recovery in click chemistry reactions. AB - The quickly developing field of "click" chemistry would undoubtedly benefit from the availability of an easy and efficient technology for product purification to reduce the potential health risks associated with the presence of copper in the final product. Therefore, solvent-resistant nanofiltration (SRNF) membranes have been developed to selectively separate "clicked" polymers from the copper catalyst and solvent. By using these solvent-stable cross-linked polyimide membranes in diafiltration, up to 98 % of the initially present copper could be removed through the membrane together with the DMF solvent, the polymer product being almost completely retained. This paper also presents the first SRNF application in which the catalyst permeates through the membrane and the reaction product is retained. PMID- 20013771 TI - Positive homotropic allosteric receptors for neutral guests: annulated tetrathiafulvalene-calix[4]pyrroles as colorimetric chemosensors for nitroaromatic explosives. AB - The study of positive homotropic allosterism in supramolecular receptors is important for elucidating design strategies that can lead to increased sensitivity in various molecular recognition applications. In this work, the cooperative relationship between tetrathiafulvalene (TTF)-calix[4]pyrroles and several nitroaromatic guests is examined. The design and synthesis of new annulated TTF-calix[4]pyrrole receptors with the goal of rigidifying the system to accommodate better nitroaromatic guests is outlined. These new derivatives, which display significant improvement in terms of binding constants, also display a positive homotropic allosteric relationship, as borne out from the sigmoidal nature of the binding isotherms and analysis by using the Hill equation, Adair equation, and Scatchard plots. The host-guest complexes themselves have been characterized by single-crystal X-ray diffraction analyses and studied by means of UV-spectroscopic titrations. Investigations into the electronic nature of the receptors were made by using cyclic voltammetry; this revealed that the binding efficiency was not strictly related to the redox potential of the receptor. On the other hand, this work serves to illustrate how cooperative effects may be used to enhance the recognition ability of TTF-calix[4]pyrrole receptors. It has led to new allosteric systems that function as rudimentary colorimetric chemosensors for common nitroaromatic-based explosives, and which are effective even in the presence of potentially interfering anions. PMID- 20013772 TI - The reaction of ozone with the hydroxide ion: mechanistic considerations based on thermokinetic and quantum chemical calculations and the role of HO4- in superoxide dismutation. AB - The reaction of OH(-) with O(3) eventually leads to the formation of *OH radicals. In the original mechanistic concept (J. Staehelin, J. Hoigne, Environ. Sci. Technol. 1982, 16, 676-681), it was suggested that the first step occurred by O transfer: OH(-)+O(3)-->HO(2)(-)+O(2) and that *OH was generated in the subsequent reaction(s) of HO(2)(-) with O(3) (the peroxone process). This mechanistic concept has now been revised on the basis of thermokinetic and quantum chemical calculations. A one-step O transfer such as that mentioned above would require the release of O(2) in its excited singlet state ((1)O(2), O(2)((1)Delta(g))); this state lies 95.5 kJ mol(-1) above the triplet ground state ((3)O(2), O(2)((3)Sigma(g)(-))). The low experimental rate constant of 70 M(-1) s(-1) is not incompatible with such a reaction. However, according to our calculations, the reaction of OH(-) with O(3) to form an adduct (OH(-)+O(3)- >HO(4)(-); DeltaG=3.5 kJ mol(-1)) is a much better candidate for the rate determining step as compared with the significantly more endergonic O transfer (DeltaG=26.7 kJ mol(-1)). Hence, we favor this reaction; all the more so as numerous precedents of similar ozone adduct formation are known in the literature. Three potential decay routes of the adduct HO(4)(-) have been probed: HO(4)(-)-->HO(2)(-)+(1)O(2) is spin allowed, but markedly endergonic (DeltaG=23.2 kJ mol(-1)). HO(4)(-)-->HO(2)(-)+(3)O(2) is spin forbidden (DeltaG=-73.3 kJ mol( 1)). The decay into radicals, HO(4)(-)-->HO(2)*+O(2)(*-), is spin allowed and less endergonic (DeltaG=14.8 kJ mol(-1)) than HO(4)(-)-->HO(2)(-)+(1)O(2). It is thus HO(4)(-)-->HO(2)*+O(2)(*-) by which HO(4)(-) decays. It is noted that a large contribution of the reverse of this reaction, HO(2)*+O(2)(*-)-->HO(4)(-), followed by HO(4)(-)-->HO(2)(-)+(3)O(2), now explains why the measured rate of the bimolecular decay of HO(2)* and O(2)(*-) into HO(2)(-)+O(2) (k=1 x 10(8) M( 1) s(-1)) is below diffusion controlled. Because k for the process HO(4)(-)- >HO(2)*+O(2)(*-) is much larger than k for the reverse of OH(-)+O(3)-->HO(4)(-), the forward reaction OH(-)+O(3)-->HO(4)(-) is practically irreversible. PMID- 20013773 TI - Bifunctional acid-base ionic liquid organocatalysts with a controlled distance between acid and base sites. AB - Bifunctional acid-base ionic liquid organocatalysts with different distances between the two sites have been synthesised, and their activity for the Knoevenagel condensation has been tested. As has been found to be the case with enzymes, the distance between the acidic and basic sites determines the activity of the bifunctional organocatalyst, and at the optimal distance the reaction rate increases by two orders of magnitude with respect to the purely acidic or basic counterpart organocatalysts. The experimental results have been rationalised through the study of the reaction mechanism of the Knoevenagel condensation between malononitrile and benzaldehyde by means of DFT calculations. It has been found that it consists of two consecutive steps. First, deprotonation of malononitrile on the basic site to obtain a methylene carbanion intermediate takes place, and second, co-adsorption and activation of benzaldehyde on the acid centre of this intermediate followed by the C-C bond-formation reaction. The calculations and the kinetic study indicate that there is an inversion of the rate-controlling step when the distance between the acidic and the basic sites is modified, with a direct implication on the reaction rate. PMID- 20013774 TI - Synthetic and structural studies on copper 1 H-[1,10]-phenanthrolin-2-one coordination complexes: isolation of a novel intermediate during 1,10 phenanthroline hydroxylation. AB - The synthesis and crystal structure elucidation of a novel dinuclear heteroleptic copper(II) complex has led to an alternative mechanism in the formation of covalent hydrates. During further studies on the synthesis and properties of [Cu(2)(ophen)(2)] (1), a dinuclear complex of copper(I) with 1 H-[1,10] phenanthrolin-2-one (Hophen), two intermediates/alternative products 2 and 3 were isolated. The dinuclear, antiferromagnetic complex [Cu(2)(ophen)(2)(phen)(2)](NO(3))(2)9H(2)O (3, phen=1,10-phenanthroline) contains two five-coordinate copper(II) ions, both with trigonal-bipyramidal coordination, which are bridged together through deprotonated hydroxyl groups with a Cu--Cu non bonding distance of 3.100 A. Complex [Cu(phen)(2)(H(2)O)](NO(3))(2) (2) is a polymorph of a previously reported material. The occurrence of 2 and 3 has led us to propose a variation to the Gillard mechanism for the formation of covalent hydrates in bidentate N-heterocycles in which the attacking nucleophile may be the deprotonated form of 2, [Cu(phen)(2)(OH)](-), rather than free OH(-). PMID- 20013775 TI - Reversible interconversion between luminescent isomeric metal-organic frameworks of [Cu(4)I(4)(DABCO)(2)] (DABCO=1,4-diazabicyclo[2.2.2]octane). AB - The metal-organic frameworks (MOF) of cluster [Cu(4)I(4)(DABCO)(2)] (DABCO=1,4 diazabicyclo[2.2.2]octane) have been prepared and characterized as two different crystalline forms, I and II. Form I is obtained by reaction of DABCO and CuI in aqueous solution or by solvothermal reaction, while form II is obtained by reacting DABCO and CuI in acetonitrile. Their luminescence properties in the solid state have been analyzed at room temperature and at 77 K. MOF II has bright emission with a maximum at 556 nm that shifts bathochromically at low temperature in conjunction with a marked change in the colour of the emission. The emission of MOF I has a maximum at 580 nm and a less pronounced temperature dependence. The peculiar luminescence properties of the two isomers have been interpreted by utilising current knowledge on the excited states properties of Cu(I) cubane clusters. The two isomers exhibit a high degree of porosity and can release the disordered solvent molecules trapped in the channels, whilst preserving the crystal structure. Isomer I can be converted into II on exposure to acetonitrile or methanol vapour, whereas II reverts to I when heated in a closed pan at 250 degrees C. PMID- 20013776 TI - Selective metal-cation recognition by [2.2]ferrocenophanes: the cases of zinc- and lithium-sensing. AB - The synthesis, electrochemical, optical, and cation-sensing properties of [2.2]ferrocenophanes, in which the two ferrocene subunits are linked through two aldiminic or iminophosphorane moieties, are reported. The new compounds show remarkably selective cation-sensing properties due to the presence of redox active units (ferrocene) and aza-unsaturated functionalities that are able to act as putative cation-binding sites. In this structural motif, the aldimine groups act as a highly selective binding site for Zn(2+) cations, whereas the iminophosphorane bridges display an unusually strong binding affinity towards Li(+) cations, which could be explained by an additional LiFe interaction. The X ray structure of the complex 4Li(+) as well as detailed NMR spectroscopic studies, both in solution and in the solid state, support this assessment. Experimental data and conclusions about the cation-sensing capabilities of this family of compounds are supported by DFT calculations. PMID- 20013777 TI - Surface dynamics of a vanadyl pyrophosphate catalyst for n-butane oxidation to maleic anhydride: an in situ raman and reactivity study of the effect of the P/V atomic ratio. AB - This work focused on investigating the effect of the P/V atomic ratio in vanadyl pyrophosphate, catalyst for n-butane oxidation to maleic anhydride, on the nature of the catalytically active phase. Structural transformations occurring on the catalyst surface were investigated by means of in situ Raman spectroscopy in a non-reactive atmosphere, as well as by means of steady-state and non-steady-state reactivity tests, in response to changes in the reaction temperature. It was found that the nature of the catalyst surface is affected by the P/V atomic ratio even in the case of small changes in this parameter. With the catalyst having P/V equal to the stoichiometric value, a surface layer made of alpha(Iota)-VOPO(4) developed in the temperature interval 340-400 degrees C in the presence of air; this catalyst gave a very low selectivity to maleic anhydride in the intermediate T range (340-400 degrees C). However, at 400-440 degrees C delta-VOPO(4) overlayers formed; at these conditions, the catalyst was moderately active but selective to maleic anhydride. With the catalyst containing a slight excess of P, the ratio offering the optimal catalytic performance, delta-VOPO(4) was the prevailing species over the entire temperature range investigated (340-440 degrees C). Analogies and differences between the two samples were also confirmed by reactivity tests carried out after in situ removal and reintegration of P. These facts explain why the industrial catalyst for n-butane oxidation holds a slight excess of P; they also explain discrepancies registered in the literature about the nature of the active layer in vanadyl pyrophosphate. PMID- 20013782 TI - Dynamics of the Dictyostelium discoideum mitochondrial proteome during vegetative growth, starvation and early stages of development. AB - In this study, a quantitative comparative proteomics approach has been used to analyze the Dictyostelium discoideum mitochondrial proteome variations during vegetative growth, starvation and the early stages of development. Application of 2-D DIGE technology allowed the detection of around 2000 protein spots on each 2 D gel with 180 proteins exhibiting significant changes in their expression level. In total, 96 proteins (51 unique and 45 redundant) were unambiguously identified. We show that the D. discoideum mitochondrial proteome adaptations mainly affect energy metabolism enzymes (the Krebs cycle, anaplerotic pathways, the oxidative phosphorylation system and energy dissipation), proteins involved in developmental and signaling processes as well as in protein biosynthesis and fate. The most striking observations were the opposite regulation of expression of citrate synthase and aconitase and the very large variation in the expression of the alternative oxidase that highlighted the importance of citrate and alternative oxidase in the physiology of the development of D. discoideum. Mitochondrial energy states measured in vivo with MitoTracker Orange CM Ros showed an increase in mitochondrial membrane polarization during D. discoideum starvation and starvation-induced development. PMID- 20013783 TI - Chemical modification of siRNA. AB - The ability to manipulate the RNA interference (RNAi) machinery to specifically silence the expression of target genes could be a powerful therapeutic strategy. Since the discovery that RNAi can be triggered in mammalian cells by short double stranded RNAs (small interfering RNA, siRNA), there has been a tremendous push by researchers, from academia to big pharma, to move siRNAs into clinical application. The challenges facing siRNA therapeutics are significant. The inherent properties of siRNAs (polyanionic, vulnerable to nuclease cleavage) make clinical application difficult due to poor cellular uptake and rapid clearance. Side effects of siRNAs have also proven to be a further complication. Fortunately, numerous chemical modification strategies have been identified that allow many of these obstacles to be overcome. This unit will present an overview of (1) the chemical modifications available to the nucleic acid chemist for modifying siRNAs, (2) the application of chemical modifications to address specific therapeutic obstacles, and (3) the factors that must be considered when assessing the activity of modified siRNAs. PMID- 20013784 TI - Nonenzymatic oligomerization of activated nucleotides on hairpin templates. AB - This unit describes a protocol for nonenzymatic oligomerization of activated ribonucleotides on DNA hairpins appended by templates containing threofuranosyl nucleic acid (TNA). TNA-cytidylate templates effectively promote oligomerization of 2-MeImpG, and give 3',5'-linked oligomerization products predominantly, with good base-pairing fidelity. Although the rates of oligomerization depend on TNA content, after 3 days of incubation, oligomerization products are apparent, and full-length products are present after 10 days. Characterization of product phosphodiester bond regiochemistry is accomplished by digestion with RNase T1. Additionally, exposure of oligomerization products to calf intestinal alkaline phosphatase enables detection of any endcapping due to pyrophosphate formation. Base-pairing fidelity is assessed by challenging the template to oligomerize 2 MeImpA. The protocols described for nonenzymatic, template-directed synthesis in this unit are applicable to oligomerization of activated monomers on templates of different compositions, with respect to both base identity and polymer backbone. PMID- 20013785 TI - Carbohydrate-oligonucleotide conjugates. AB - This unit describes a strategy for attaching two mannose and two galactose residues to an oligonucleotide. This conjugation can be performed at the 5'-end of the oligonucleotide sequence, using modified phosphoramidites. First, the oligonucleotide scaffold is synthesized on solid support using a DNA synthesizer, with commercially available and modified phosphoramidites. After the first "click" reaction with a galactosylated azide derivative on solid support, the bromine atoms are replaced with azides and a second click reaction is performed with propargylated mannose either on solid support or in solution. Additionally, using a monoalkynated solid support, the conjugation with carbohydrate residues can be performed at the 3'-end of the oligonucleotide according to a similar protocol. Curr. Protoc. Nucleic Acid Chem. 39:4.38.1-4.38.25. PMID- 20013786 TI - Design, synthesis, and amplification of DNA pools for in vitro selection. AB - Preparation of a random-sequence DNA pool is presented. The degree of randomization and the length of the random sequence are discussed, as is synthesis of the pool using a DNA synthesizer or via commercial synthesis companies. Purification of a single-stranded pool and conversion to a double stranded pool are presented as step-by-step protocols. Support protocols describe determination of the complexity and skewing of the pool, and optimization of amplification conditions. PMID- 20013787 TI - The clinical relevance of Wilms Tumour 1 (WT1) gene mutations in acute leukaemia. AB - Recurrent genetic aberrations are important predictors of outcome in acute myeloid leukaemia (AML). Numerous novel molecular abnormalities have been identified and investigated in recent years adding to the risk stratification and prognostication of conventional karyotyping. Mutations in the Wilms Tumour 1 (WT1) gene were first described more than a decade ago but their clinical significance has only recently been evaluated. WT1 mutations occur in approximately 10% of adult AML patients at diagnosis and are most frequent in the cytogenetically normal (CN) AML subgroup. These mutations appear to confer a negative prognostic outcome by increasing the risk of relapse and death. Mutation frequency is higher in pediatric patients and also appears to confer a negative impact on relapse and survival. Herein, we discuss the importance of WT1 mutations in AML. PMID- 20013788 TI - Establishment and characteristics of porcine preadipocyte cell lines derived from mature adipocytes. AB - Development of established preadipocyte cell lines, such as 3T3-L1 and 3T3-F442A, greatly facilitated the study of molecular mechanisms of adipocyte differentiation under defined conditions. Most of these cell lines are derived from mouse embryos, and preadipocyte cell lines of other species have not yet been maintained in culture long enough to study differentiation under a variety of conditions. This is the first report on the establishment of porcine preadipocyte cell lines derived from mature adipocytes by a simple method, known as ceiling culture, for culturing mature adipocytes in vitro. This cell line can proliferate extensively until the cells become confluent and fully differentiated into mature adipocytes, depending on adipogenic induction. No changes in their differentiation pattern are observed during their propagation, and they have been successfully carried and differentiated for at least 37 passages. This cell line maintains a normal phenotype without transforming spontaneously, even after long term maintenance in culture. This achievement may lead to easy establishment of porcine preadipocyte cell lines and novel model systems for studying the mechanisms of adipocyte differentiation and metabolism as a substitute for human preadipocytes. PMID- 20013790 TI - Comparative skull osteology of Karsenia koreana (Amphibia, Caudata, Plethodontidae). AB - The recent discovery of a plethodontid salamander, Karsenia koreana, in Korea challenged our understanding of the biogeographic history of the family Plethodontidae, by far the largest family of salamanders, which otherwise is distributed in the New World with a few European species. Molecular studies suggest that Karsenia forms a clade with Hydromantes (sensu lato), which includes among its species the only other Old World plethodontids. We studied the skull of K. koreana and compared it with that of other plethodontid genera, especially members of the subfamily Plethodontinae, which it resembles most closely in general anatomy. The anatomy of its skull corresponds to the most generalized and apparently ancestral condition for plethodontids. No clearly autapomorphic states were detected, and no synapomorphies can be found that would link it to other genera. The Karsenia skull is cylindrical and well ossified, giving an impression of strength. In contrast, the skull of Hydromantes is highly derived; the skull is flattened and the bones are weakly ossified and articulated. Hydromantes and Karsenia share no unique anatomical features; differences between them are especially evident in the hyobranchial skeleton, which is generalized in Karsenia but highly modified in Hydromantes, which is well known for its highly projectile tongue. Plethodon and Plethodon-like species, including Karsenia and to a lesser degree Ensatina, represent the more generalized and apparently ancestral plethodontid morphology. Specialized morphologies have evolved along only a few morphological axes within the Plethodontidae, resulting in a pattern of rampant homoplasy. Our analysis of the anatomy of the new Asiatic lineage illuminates some potential mechanisms underlying adaptive morphological evolution within the Plethodontidae. PMID- 20013789 TI - Posthatching development of Alligator mississippiensis ovary and testis. AB - We investigated ovary and testis development of Alligator mississippiensis during the first 5 months posthatch. To better describe follicle assembly and seminiferous cord development, we used histochemical techniques to detect carbohydrate-rich extracellular matrix components in 1-week, 1-month, 3-month, and 5-month-old gonads. We found profound morphological changes in both ovary and testis. During this time, oogenesis progressed up to diplotene arrest and meiotic germ cells increasingly interacted with follicular cells. Concomitant with follicles becoming invested with full complements of granulosa cells, a periodic acid Schiff's (PAS)-positive basement membrane formed. As follicles enlarged and thecal layers were observed, basement membranes and thecal compartments gained periodic acid-methionine silver (PAMS)-reactive fibers. The ovarian medulla increased first PAS- and then PAMS reactivity as it fragmented into wide lacunae lined with low cuboidal to squamous epithelia. During this same period, testicular germ cells found along the tubule margins were observed progressing from spermatogonia to round spermatids located within the center of tubules. Accompanying this meiotic development, interstitial Leydig cell clusters become more visible and testicular capsules thickened. During the observed testis development, the thickening tunica albuginea and widening interstitial tissues showed increasing PAS- and PAMS reactivity. We observed putative intersex structures in both ovary and testis. On the coelomic aspect of testes were cell clusters with germ cell morphology and at the posterior end of ovaries, we observed "medullary rests" resembling immature testis cords. We hypothesize laboratory conditions accelerated gonad maturation due to optimum conditions, including nutrients and temperature. Laboratory alligators grew more rapidly and with increased body conditions compared with previous measured, field-caught animals. Additionally, we predict the morphological maturation observed in these gonads is concomitant with increased endocrine activities. PMID- 20013791 TI - The ontogeny of Pseudis platensis (Anura, Hylidae): Heterochrony and the effects of larval development on postmetamorphic life. AB - Recent studies have described the giant tadpole, delayed metamorphic transformations, and absence of postmetamorphic growth of the skeleton of Pseudis Platensis. These features address questions about derived patterns of life cycles and the role of the heterochrony during the metamorphosis in anurans. Using anatomical methods, we provide new data on the development of reproductive, digestive and integument systems, and age inference obtained from ontogenetic series of Pseudis platensis. Our results indicate that at the end of metamorphosis, the adult skin is completely differentiated, including the calcified dermal layer; the testis has seminiferous tubules with spermatogonia, spermatocytes, and spermatids; ovarian sacs present previtellogenic ova; and the adult digestive tract is fully formed. The froglets differ from adults only in being unable to reproduce. The entire life cycle of P. platensis can occur in 4 years. In the first year, larval development, growth to adult size, and gonad differentiation are completed. Long larval development rather than size of the tadpoles seems to be involved in the absence of juvenile stages. PMID- 20013792 TI - Ultrastructure of the extensively developed nuchal organs of Laonice bahusiensis (Annelida: Canalipalpata: Spionidae). AB - The nuchal organs of annelid Laonice bahusiensis (Spionidae) from northern Europe have been studied using scanning and transmission electron microscopy. L. bahusiensis is the first spionid species in which extensively developed, continuous nuchal organs are described. The nuchal organs of this genus are the longest known among polychaete annelids. They consist of paired double bands extending from the prostomium on a mid-dorsal caruncle for about 24-30 setigers. Their microanatomy corresponds to the general structural plan of nuchal organs: there are ciliated supporting cells and bipolar sensory cells with sensory cilia traversing an olfactory chamber. The organs are overlaid by a secondary paving stone-like cover and innervated by one pair of longitudinally elongated nuchal nerves. These findings clearly favor the hypothesis that the paired, extensively developed ciliated structures found in some Spionidae are homologous with the prostomial nuchal organs characteristic of polychaete annelids. PMID- 20013793 TI - An inventory and update of jealousy-evoking partner behaviours in modern society. AB - The goal of the present study was to identify the most important jealousy-evoking partner behaviours and to examine the extent to which these behaviours evoke jealousy. Based on the literature, a questionnaire was constructed containing 42 jealousy-evoking partner behaviours, including a partner's extra-dyadic involvement with someone else by means of modern communication devices, such as the Internet. A second study examined the extent to which undergraduates and a community sample experienced jealousy in response to these partner behaviours. Results showed that explicit unfaithful behaviours evoked most feelings of jealousy, followed by a partner's emotional or romantic involvement with someone else by means of modern communication devices. In general, older individuals responded with less jealousy in response to a partner's unfaithful and suspicious behaviours. Clinical implications are discussed. PMID- 20013794 TI - Endomyocardial biopsy derived adherent proliferating cells - a potential cell source for cardiac tissue engineering. AB - Heart diseases are a leading cause of morbidity and mortality. Cardiac stem cells (CSC) are considered as candidates for cardiac-directed cell therapies. However, clinical translation is hampered since their isolation and expansion is complex. We describe a population of human cardiac derived adherent proliferating (CAP) cells that can be reliably and efficiently isolated and expanded from endomyocardial biopsies (0.1 cm(3)). Growth kinetics revealed a mean cell doubling time of 49.9 h and a high number of 2.54 x 10(7) cells in passage 3. Microarray analysis directed at investigating the gene expression profile of human CAP cells demonstrated the absence of the hematopoietic cell markers CD34 and CD45, and of CD90, which is expressed on mesenchymal stem cells (MSC) and fibroblasts. These data were confirmed by flow cytometry analysis. CAP cells could not be differentiated into adipocytes, osteoblasts, chondrocytes, or myoblasts, demonstrating the absence of multilineage potential. Moreover, despite the expression of heart muscle markers like alpha-sarcomeric actin and cardiac myosin, CAP cells cannot be differentiated into cardiomyocytes. Regarding functionality, CAP cells were especially positive for many genes involved in angiogenesis like angiopoietin-1, VEGF, KDR, and neuropilins. Globally, principal component and hierarchical clustering analysis and comparison with microarray data from many undifferentiated and differentiated reference cell types, revealed a unique identity of CAP cells. In conclusion, we have identified a unique cardiac tissue derived cell type that can be isolated and expanded from endomyocardial biopsies and which presents a potential cell source for cardiac repair. Results indicate that these cells rather support angiogenesis than cardiomyocyte differentiation. PMID- 20013795 TI - S100A6 (calcyclin) deficiency induces senescence-like changes in cell cycle, morphology and functional characteristics of mouse NIH 3T3 fibroblasts. AB - S100A6 (calcyclin) is a calcium binding protein with two EF-hand structures expressed mostly in fibroblasts and epithelial cells. We have established a NIH 3T3 fibroblast cell line stably transfected with siRNA against S100A6 to examine the effect of S100A6 deficiency on non-transformed cell physiology. We found that NIH 3T3 fibroblasts with decreased level of S100A6 manifested altered cell morphology and proliferated at a much slower pace than the control cells. Cell cycle analysis showed that a large population of these cells lost the ability to respond to serum and persisted in the G0/G1 phase. Furthermore, fibroblasts with diminished S100A6 level exhibited morphological changes and biochemical features of cellular senescence as revealed by beta-galactosidase and gelatinase assays. Also, S100A6 deficiency induced changes in the actin cytoskeleton and had a profound impact on cell adhesion and migration. Thus, we have shown that the S100A6 protein is involved in multiple aspects of fibroblast physiology and that its presence ensures normal fibroblast proliferation and function. PMID- 20013796 TI - FIAT control of osteoblast activity. AB - The basic domain-leucine zipper transcription factor activating transcription factor 4 (ATF4) regulates most functions of the osteoblast. It is therefore not surprising that its activity should be regulated through several mechanisms. Factor inhibiting ATF4-mediated transcription (FIAT) is a leucine zipper nuclear molecule lacking a basic domain for DNA binding that interacts with ATF4 to repress its transcriptional activity. FIAT expression was monitored in parallel with ATF4 during osteoblastogenesis. The mechanism of ATF4 repression by FIAT was investigated through structure-function analysis. The physiological significance of FIAT expression in osteoblasts was studied through silencing FIAT in osteoblasts by RNA interference, as well as through characterization of two genetic mouse models: FIAT transgenic mice which overexpress FIAT in osteoblasts, and FIAT knockout mice. Studies to date show that FIAT and ATF4 are co-expressed in osteoblasts, and that FIAT inhibition of matrix mineralization requires dimerization with ATF4 through the second leucine zipper. Furthermore, transgenic mice overexpressing FIAT exhibit osteopenia. The phenotype of FIAT knockout mice is still under evaluation but the salient aspects are discussed here. Taken together, the results accumulated to date support the hypothesis that FIAT is a transcriptional repressor that modulates osteoblast function. PMID- 20013797 TI - A single deletion at position 134, 135, or 136 in the beta 7-beta 8 loop of the p51 subunit of HIV-1 RT disrupts the formation of heterodimeric enzyme. AB - The human immunodeficiency virus type 1 reverse transcriptase (HIV-1 RT) is a heterodimeric enzyme composed of p66 and p51 subunits. Earlier, we showed that the beta7-beta8 loop of p51 is crucial for polymerase activity of HIV-1 RT as either deletion or Ala substitution of amino acids in the beta7-beta8 loop spanning residues 136-139 in the p51 subunit impaired dimerization and, in turn, polymerase function of the enzyme (Pandey et al. 2001 Biochemistry 40: 9505 9512). In the present study, we generated subunit-specific single-deletion mutants at positions 134, 135, 136, or 137 and examined their effects on the heterodimerization, binary complex formation, and polymerase functions of the enzyme. We found that among these four residues, Ser134, Ile135, and Asn136 in the beta7-beta8 loop of the p51 subunit are crucial residues for dimerization and polymerase function of the enzyme, but have no impact when specifically deleted from the p66 subunit. These results demonstrate the beta7-beta8 loop of the p51 subunit in the formation of stable, functional heterodimeric enzyme which could be an attractive target for anti-HIV-1 drug development. PMID- 20013798 TI - Anisotropy of spin relaxation of water protons in cartilage and tendon. AB - Transverse spin relaxation rates of water protons in articular cartilage and tendon depend on the orientation of the tissue relative to the applied static magnetic field. This complicates the interpretation of magnetic resonance images of these tissues. At the same time, relaxation data can provide information about their organisation and microstructure. We present a theoretical analysis of the anisotropy of spin relaxation of water protons observed in fully hydrated cartilage. We demonstrate that the anisotropy of transverse relaxation is due almost entirely to intramolecular dipolar coupling modulated by a specific mode of slow molecular motion: the diffusion of water molecules in the hydration shell of a collagen fibre around the fibre, such that the molecular director remains perpendicular to the fibre. The theoretical anisotropy arising from this mechanism follows the 'magic-angle' dependence observed in magnetic-resonance measurements of cartilage and tendon and is in good agreement with the available experimental results. We discuss the implications of the theoretical findings for MRI of ordered collagenous tissues. PMID- 20013799 TI - Proteomic characterization of a pilR regulatory mutant of Azoarcus sp. strain BH72 with the aid of gel-based and gel-free approaches. AB - The proteome of the grass endophyte Azoarcus sp. strain BH72 was analyzed by a combination of gel-based methods by means of 2-DE and MS and a gel-free approach via LC-MS/MS. Among the identified 785 proteins, synthesis of around 100 conserved hypothetical proteins could be confirmed. Membrane proteins were detected at a higher rate in the gel-free than in the gel-based approach. The abundance of proteins in the constructed proteome reference map was analyzed and the 30 most abundant proteins were determined. The reference map was then used as a starting point to characterize the regulon under control of the response regulator PilR. PilR is part of the two-component regulatory system PilSR controlling type IV pilin gene (pilAB) expression in strain BH72, which was strongly decreased (19.7-fold) in the pilR-mutant. Changes of protein composition in the wild type and the regulatory mutant were compared by the gel-based and gel free analyses. Proteins responsible for amino acid and energy metabolism, chaperones as well as proteins that are involved in iron metabolism and iron storage were present in a pilR-mutant at different levels than in the wild-type strain. Levels of the transcriptional regulator Flp were also dependent on PilR, indicating that PilR might be part of a hierarchical regulatory cascade. PMID- 20013800 TI - Modulation of cellulosome composition in Clostridium cellulolyticum: adaptation to the polysaccharide environment revealed by proteomic and carbohydrate-active enzyme analyses. AB - Clostridium cellulolyticum is a model mesophilic anaerobic bacterium that efficiently degrades plant cell walls. The recent genome release offers the opportunity to analyse its complete degradation system. A total of 148 putative carbohydrate-active enzymes were identified, and their modular structures and activities were predicted. Among them, 62 dockerin-containing proteins bear catalytic modules from numerous carbohydrate-active enzymes' families and whose diversity reflects the chemical and structural complexity of the plant carbohydrate. The composition of the cellulosomes produced by C. cellulolyticum upon growth on different substrates (cellulose, xylan, and wheat straw) was investigated by LC MS/MS. The majority of the proteins encoded by the cip-cel operon, essential for cellulose degradation, were detected in all cellulosome preparations. In the presence of wheat straw, the natural and most complex of the substrates studied, additional proteins predicted to be involved in hemicellulose degradation were produced. A 32-kb gene cluster encodes the majority of these proteins, all harbouring carbohydrate-binding module 6 or carbohydrate-binding module 22 xylan-binding modules along dockerins. This newly identified xyl-doc gene cluster, specialised in hemicellulose degradation, comes in addition of the cip-cel operon for plant cell wall degradation. Hydrolysis efficiencies determined on the different substrates corroborates the finding that cellulosome composition is adapted to the growth substrate. PMID- 20013801 TI - Mesoporous silica chips for selective enrichment and stabilization of low molecular weight proteome. AB - The advanced properties of mesoporous silica have been demonstrated in applications, which include chemical sensing, filtration, catalysis, drug delivery and selective biomolecular uptake. These properties depend on the architectural, physical and chemical properties of the material, which in turn are determined by the processing parameters in evaporation-induced self-assembly. In this study, we introduce a combinatorial approach for the removal of the high molecular weight proteins and for the specific isolation and enrichment of low molecular weight species. This approach is based on mesoporous silica chips able to fractionate, selectively harvest and protect from enzymatic degradation, peptides and proteins present in complex human biological fluids. We present the characterization of the harvesting properties of a wide range of mesoporous chips using a library of peptides and proteins standard and their selectivity on the recovery of serum peptidome. Using MALDI-TOF-MS, we established the correlation between the harvesting specificity and the physicochemical properties of mesoporous silica surfaces. The introduction of this mesoporous material with fine controlled properties will provide a powerful platform for proteomics application offering a rapid and efficient methodology for low molecular weight biomarker discovery. PMID- 20013803 TI - Expression and prognostic significance of APAF-1, caspase-8 and caspase-9 in stage II/III colon carcinoma: caspase-8 and caspase-9 is associated with poor prognosis. AB - Apoptosis protease activating factor-1 (APAF-1), caspase-8 and caspase-9 are important factors in the execution of death signals. To study their prognostic influence in colon carcinoma, expression of APAF-1, caspase-8 and caspase-9 was determined by immunohistochemistry in normal colon mucosa (n = 8) and R0-resected stage II/III colon carcinomas (n >or= 124) using a semiquantitative score. Staining results were correlated with disease-free survival by Kaplan-Meier estimates, and multivariate Cox analyses were performed. In normal colon, APAF-1 and caspase-8 are most strongly expressed in the luminal surface epithelium, whereas caspase-9 is expressed all along the crypt axis. In colon carcinomas, there is considerable variability in the expression of these proapoptotic factors, although complete loss of caspase-8 and caspase-9 is rare. APAF-1 expression did not correlate with disease-free survival. Instead, both expression of caspase-9 and high-level expression of caspase-8 in a majority of tumor cells were significantly associated with adverse prognosis (p = 0.004 and p = 0.029, respectively). The influence of caspase-8 expression was mainly seen in patients with stage III colon carcinoma (p = 0.011), whereas the prognostic influence of caspase-9 expression was significant in stage II cases (p = 0.037) and just failed to be significant in stage III tumors (p = 0.0581). After adjusting for confounding factors in a multivariate Cox analysis, the effect of caspase-9 in predicting disease-free survival was confirmed (p = 0.003). Our data suggest that, in colon carcinomas, expression of caspase-8 and caspase-9 is significantly associated with poor survival. Caspase-9 may be an independent prognosticator in colon carcinoma. PMID- 20013802 TI - Overexpression of DDB2 enhances the sensitivity of human ovarian cancer cells to cisplatin by augmenting cellular apoptosis. AB - Cisplatin is one of the most widely used anticancer agents, displaying activity against a wide variety of tumors. However, development of drug resistance presents a challenging barrier to successful cancer treatment by cisplatin. To understand the mechanism of cisplatin resistance, we investigated the role of damaged DNA binding protein complex subunit 2 (DDB2) in cisplatin-induced cytotoxicity and apoptosis. We show that DDB2 is not required for the repair of cisplatin-induced DNA damage, but can be induced by cisplatin treatment. DDB2 deficient noncancer cells exhibit enhanced resistance to cell growth inhibition and apoptosis induced by cisplatin than cells with fully restored DDB2 function. Moreover, DDB2 expression in cisplatin-resistant ovarian cancer cell line CP70 and MCP2 was lower than their cisplatin-sensitive parental A2780 cells. Overexpression of DDB2 sensitized CP70 cells to cisplatin-induced cytotoxicity and apoptosis via activation of the caspase pathway and downregulation of antiapoptotic Bcl-2 protein. Further analysis indicates that the overexpression of DDB2 in CP70 cells downregulates Bcl-2 expression through decreasing Bcl-2 mRNA level. These results suggest that ovarian cancer cells containing high level of DDB2 become susceptible to cisplatin by undergoing enhanced apoptosis. PMID- 20013804 TI - E2F-1 RNomics is critical for reprogramming of cancer cells to quiescent state. AB - The discovery of cooperativity between pRB and E2F greatly prompted various investigators to find how E2F biology contributes to oncogenesis. Although E2F family of transcription factors have been linked to proliferation, apoptosis and differentiation, yet no heed has been paid to understand the role of E2F biology in cellular quiescence. To understand the functional RNomics (regulation of gene transcription through RNA interference) of E2F-1 gene, 2 cancer cell lines, such as Jurkat exhibiting E2F-1 gene overamplification and Hela-229 exhibiting intrinsic downregulation of E2F-1 gene expression, were used in our study. E2F-1 gene knockdown via siRNA within Jurkat cells resulted in upregulation of genes characteristic of quiescence both translationally and transcriptionally, which was accompanied by downregulation of genes at both translational and transcriptional level involved in cell cycle progression and apoptosis. This genomic phenomenon also translated into ultrastructural and phenotypic features typical of quiescent state. These observed results in Jurkat cells were simulated by upregulation of E2F-1 gene in Hela-229 cells through the downregulation of miR 17-5p. This E2F-1-regulated pathway explained as to how Jurkat cells entered exclusively into quiescent state when E2F-1 was downregulated in these cells and how Hela-229 cells proliferate vigorously when E2F-1 was upregulated in such cells. Here, we propose a gene-regulatory pathway which the cell might be using in its entry into either quiescent or proliferative states. Furthermore, this pathway may be useful in designing strategies for the treatment of cancer in general and acute lymphoblastic leukemia in particular. PMID- 20013805 TI - A novel role of gap junction connexin46 protein to protect breast tumors from hypoxia. AB - Connexin proteins are the principle structural components of the gap junctions. Colocalization and tissue-specific expression of diverse connexin molecules are reported to occur in a variety of organs. Impairment of gap junctional intercellular communication, caused by mutations, gain of function or loss of function of connexins, is involved in a number of diseases including the development of cancer. Here we show that human breast cancer cells, MCF-7 and breast tumor tissues express a novel gap junction protein, connexin46 (Cx46) and it plays a critical role in hypoxia. Previous studies have shown that connexin46 is predominantly expressed in lens and our studies find that Cx46 protects human lens epithelial cells from hypoxia induced death. Interestingly, we find that Cx46 is upregulated in MCF-7 breast cancer cells and human breast cancer tumors. Downregulation of Cx46 by siRNA promotes 40% MCF-7 cell death at 24 hr under hypoxic conditions. Furthermore, direct injection of anti-Cx46 siRNA into xenograft tumors prevents tumor growth in nude mice. This finding will provide an exciting new direction for drug development for breast cancer treatment and suggests that both normal hypoxic tissue (lens) and adaptive hypoxic tissue (breast tumor) utilize the same protein, Cx46, as a protective strategy from hypoxia. PMID- 20013806 TI - Lack of HLA class II antigen expression in microsatellite unstable colorectal carcinomas is caused by mutations in HLA class II regulatory genes. AB - Colorectal cancers (CRCs) develop on the basis of a deficient DNA mismatch repair (MMR) system in about 15% of cases. MMR-deficient CRC lesions show high-level microsatellite instability (MSI-H) and accumulate numerous mutations located at coding microsatellite loci that lead to the generation of immunogenic neopeptides. Consequently, the host's antitumoral immune response is of high importance for the course of the disease in MSI-H CRC patients. Accordingly, immune evasion mediated by impairment of HLA class I antigen presentation is frequently observed in these cancers. In this study, we aimed at a systematic analysis of alterations affecting HLA class II antigen expression in MSI-H CRC. HLA class II antigens are expressed by only two-thirds of MSI-H CRCs. The mechanisms underlying the lack of HLA class II antigens in a subset of MSI-H CRCs remain unknown. We here screened HLA class II regulatory genes for the presence of coding microsatellites and identified mutations of the essential regulator genes RFX5 in 9 (26.9%) out of 34 and CIITA in 1 (2.9%) out of 34 MSI-H CRCs. RFX5 mutations were related to lack of or faint HLA class II antigen expression (p = 0.006, Fisher's exact test). Transfection with wild-type RFX5 was sufficient to restore interferon gamma-inducible HLA class II antigen expression in the RFX5 mutant cell line HDC108. We conclude that somatic mutations of the RFX5 gene represent a novel mechanism of loss of HLA class II antigen expression in tumor cells, potentially contributing to immune evasion in MSI-H CRCs. PMID- 20013807 TI - Anti-inflammatory M2 type macrophages characterize metastasized and tyrosine kinase inhibitor-treated gastrointestinal stromal tumors. AB - We have made a detailed inventory of the immune infiltrate of gastrointestinal stromal tumors (GISTs), which originate from mesenchymal cells in the intestinal tract. These sarcomas are heavily infiltrated with macrophages and T cells, while immune cells of other lineages were much less abundant. Dissecting the functional subtypes of T cells with multicolor fluorescent microscopy revealed substantial populations of cytotoxic T cells, helper T cells and FoxP3(+) regulatory T cells. The balance of cytotoxic T cells and FoxP3(+) T cells was toward immune suppression. Analysis of the macrophage population also showed a dominance of anti-inflammatory cells, as the M2 type scavenger receptor CD163 was abundantly present. Other subsets of macrophages (CD14(+)CD163(-)) were occasionally detected. M2 type CD163(+) macrophages were associated with the number of infiltrating FoxP3(+) regulatory T cells and twice as many macrophages were found in metastatic GIST compared to primary lesions. Most metastatic GISTs had been treated with the tyrosine kinase inhibitors imatinib and sunitinib, but the high macrophage infiltrate was not related to this treatment. However, imatinib and sunitinib did induce secretion of anti-inflammatory IL-10 in macrophage cultures, indicating that treatment with these inhibitors might contribute to an immune suppressive microenvironment in GIST. Overall, our data reveal a picture of GIST as an active site of tumor-immune interaction in which suppressive mechanisms overrule potential antitumor responses. Tyrosine kinase inhibitors might promote this negative balance. PMID- 20013808 TI - Antitumorigenic effects of a mutant of the heparin affin regulatory peptide on the U87 MG glioblastoma cell line. AB - Glioblastoma is the most common primary brain tumor in human adults. Since existing treatments are not effective enough, novel therapeutic targets must be sought. The heparin-binding growth factor, heparin affin regulatory peptide (HARP), also known as pleiotrophin (PTN), could potentially represent such a target. We have previously shown that a mutant protein, HARPDelta111-136, which lacks HARP's C-terminal 26 amino acids, acts as a dominant negative HARP effector by heterodimerizing with the wild-type growth factor. The aim of our study was to evaluate the potential inhibitory activity of HARPDelta111-136 on the U87 MG human glioblastoma cell line. By overexpressing the truncated form of HARP in stably established clones of U87 MG cells, we observed an inhibition of proliferation under both anchorage-dependent and anchorage-independent conditions. We confirmed these results in an in vivo subcutaneous tumor xenograft model. In addition, we found that HARPDelta111-136 inhibited cell proliferation in a paracrine manner. Analysis of key cellular pathways revealed a decrease of cell adhesion in U87 MG cells that overexpressed the mutant protein, which could explain this inhibitory effect. A replication-defective adenovirus model that encoded HARPDelta111-136 supported a putative antiproliferative role for the truncated protein in vitro and in vivo. Interestingly, HARPDelta111-136 was also able to abolish angiogenic activity in HUVEC proliferation and in a Matrigel plug assay. These results demonstrate that considering its antiproliferative and angiostatic effects, HARPDelta111-136 could be of great interest when used in conjunction with standard treatments. PMID- 20013809 TI - Suppression of tumorigenesis and metastasis of hepatocellular carcinoma by shRNA interference targeting on homeoprotein Six1. AB - We previously demonstrated that the overexpression of homeoprotein Six1 in hepatocellular carcinoma (HCC) patients is associated with venous infiltration, advanced pathologic tumor metastasis (pTNM) stage and poor overall survival rate (Ng et al. Br J Cancer 2006;95:1050-5). In this study, short hairpin RNA (shRNA) interference approach was used to suppress the expression of Six1 in a metastatic HCC cell line MHCC97L. Stable transfectant MHCC97L-shSix1 carrying Six1-specific shRNA plasmid was established to downregulate Six1 expression to about 40% when compared with MHCC97L-Control. In vitro functional assays demonstrated that the growth rate and proliferation ability of MHCC97L-shSix1 cells were markedly decreased. Moreover, significant decrease of cell motility and invasiveness were observed in MHCC97L-shSix1 cells. Data from in vivo xenograft tumorigenesis model demonstrated that the size of tumor in MHCC97L-shSix1 group was dramatically reduced. Experimental and spontaneous metastasis models indicated that targeting Six1 suppression noticeably reduced the pulmonary metastasis in MHCC97L-shSix1 group. To identify Six1-regulated targets, cDNA microarray was employed to compare the expression profiles of MHCC97L-Control and MHCC97L-shSix1 cells. Twenty-eight downregulated and 24 upregulated genes with known functions were identified in MHCC97L-shSix1. The functions of these target genes are involved in diverse biological activities. Our data suggest that Six1 may be involved in regulation of proliferation and invasiveness of HCC; thus targeting suppression of Six1 is a viable option for treating HCC patients. PMID- 20013810 TI - Nerve invasion distance is dependent on laminin gamma2 in tumors of pancreatic cancer. AB - The distance of nerve invasion is an important prognostic factor in pancreatic cancer. The extracellular matrix (ECM) of nerve, mainly composed of laminin, collagen IV and anchoring fibrils, might affect nerve invasion. However, this relationship has not been demonstrated. Our study aimed at discovering the promoting factor of nerve invasion within the tumoral ECM. An animal model was established to evaluate the distance of nerve invasion in murine sciatic nerves by intraneural injection of 6 human pancreatic cancer cell lines. mRNA expression of laminins and anchoring fibrils was compared to the distance of nerve invasion for each cancer cell line. A target molecule provided the strong association between mRNA expression and the distance of nerve invasion. To evaluate the role of a target molecule in nerve invasion, protein expression and function were examined using an animal model and surgical cases. Cancer cells with high laminin gamma2 mRNA and protein expression in their basement membranes were associated with long nerve invasion. Knockdown of laminin gamma2 in cancer cells significantly shortened nerve invasion in the animal model. In 75 patients with pancreatic cancer, a large distance of nerve invasion was associated with high expression levels of laminin gamma2 mRNA and basement membranous deposition of laminin gamma2 protein. Our results indicate that laminin gamma2 plays an important role in nerve invasion. The measurement of the nerve invasion distance in our mouse nerve invasion model is useful for evaluating the molecular mechanisms of nerve invasion. PMID- 20013811 TI - The precancerous effect of emitted cooking oil fumes on precursor lesions of cervical cancer. AB - Although cooking emission from high-temperature frying has been deemed a Group 2A carcinogen by the International Agency for Research on Cancer, little is known about its impact on cervical tumorigenesis. To investigate the precancerous consequence of cooking oil fumes on cervical intraepithelial neoplasm (CIN), a community-based case-control study, which takes all known risk factors into consideration, was conducted in Taiwan. From 2003 to 2008, in a Pap smear screening and biopsy examination network, 206 pathology-verified women with inflammations/atypical squamous cells of undetermined significance or CIN grade-1 (CIN1) and 73 with CIN2-3 (defined as low-grade squamous intraepithelial lesions (LGSIL) and high-grade squamous intraepithelial lesions (HGSIL), respectively); and 1,200 area-and-age-matched controls with negative cytology were recruited. Multinomial logistic regression was applied in the multivariate analysis to determine the likelihood of contracting LGSIL or HGSIL. The risks of the two lesions increased with the increase of carcinogenic high-risk human papillomavirus DNA load, with a clear dose-response relationship. Chefs were observed to experience a 7.9-fold elevated HGSIL risk. Kitchens with poor fume ventilation during the main cooking life-stage correlated to a 3.7-fold risk of HGSIL, but not for LGSIL. More than 1 hr of daily cooking in kitchens with poor fume conditions appeared to confer an 8.4-fold HGSIL risk, with an 8.3-fold heterogeneously higher odds ratio than that (aOR = 1.0) for LGSIL. Similar risk pattern has been reproduced among never-smoking women. Our findings demonstrate the association between indoor exposure to cooking fumes from heated oil and the late development of cervical precancerous lesions. This final conclusion needs to be verified by future research. PMID- 20013812 TI - Establishing regiocontrol of disulfide bond isomers of alpha-conotoxin ImI via the synthesis of N-to-C cyclic analogs. AB - alpha-Conotoxins are multiple disulfide bond containing peptides that are isolated from venomous marine cone snails. They display remarkable selectivity for different subtypes of nicotinic acetylcholine receptors (nAChRs). While alpha conotoxins display poor resistance to in vivo degradation by proteases, which limits their use as drug leads, N-to-C cyclization via an oligopeptide spacer unit has been previously shown to improve stability. However, the effect of N-to C cyclization on the formation of the disulfide bond framework is not fully understood. Four N-to-C cyclic analogs of alpha-conotoxin ImI; cImI-A, cImI betaA, cImI-AG, and cImI-AGG were synthesized to evaluate the effect of oligopeptide spacer length on disulfide bond selectivity and stability to proteolysis. Different ratios of disulfide bond isomers were obtained for each analog using a nonselective random disulfide bond forming strategy, which was dependent on the length of the spacer. To identify each isomer obtained using the random strategy, and to gain access to disulfide bond isomers otherwise unattainable using the random strategy, both the native (globular) and ribbon isomers were synthesized in good yield and purity using a selective orthogonal cysteine protecting group strategy. As such, a random oxidation strategy showed a clear preference for the ribbon isomer in cImI-A. The cyclic globular isomers showed a high resistance to enzymatic degradation compared to the ribbon isomers, with the cImI-A and cImI-AG globular isomers demonstrating the highest stability. These results suggest that cyclization can improve the biochemical stability of conotoxins with potential applications in the development of drugs. PMID- 20013813 TI - Effects of 1alpha,25-dihydroxyvitamin D3 on transporters and enzymes of the rat intestine and kidney in vivo. AB - 1alpha,25-Dihydroxyvitamin D3 (1,25(OH)2D3), the natural ligand of the vitamin D receptor (VDR), was found to regulate bile acid related transporters and enzymes directly and indirectly in the rat intestine and liver in vivo. The kidney is another VDR-rich target organ in which VDR regulation on xenobiotic transporters and enzymes is ill-defined. Hence, changes in protein and mRNA expression of nuclear receptors, transporters and enzymes of the rat intestine and kidney in response to 1,25(OH)2D3 treatment (0 to 2.56 nmol/kg/day intraperitoneally in corn oil for 4 days) were studied. In the intestine, protein and not mRNA levels of Mrp2, Mrp3, Mrp4 and PepT1 in the duodenum and proximal jejunum were induced, whereas Oat1 and Oat3 mRNA were decreased in the ileum after 1,25(OH)2D3 treatment. In the kidney, VDR, Cyp24, Asbt and Mdr1a mRNA and protein expression increased significantly (2- to 20-fold) in 1,25(OH)2D3-treated rats, and a 28 fold increase of Cyp3a9 mRNA but not of total Cy3a protein nor Cyp3a1 and Cyp3a2 mRNA was observed, implicating that VDR played a significant, renal-specific role in Cyp3a9 induction. Additionally, renal mRNA levels of PepT1, Oat1, Oat3, Ostalpha, and Mrp4, and protein levels of PepT1 and Oat1 were decreased in a dose dependent manner, and the approximately 50% concomitant reduction in FXR, SHP, HNF-1alpha and HNF-4alpha mRNA expression suggests the possibility of cross-talk among the nuclear receptors. It is concluded that the effects of 1,25(OH)2D3 changes are tissue-specific, differing between the intestine and kidney which are VDR-rich organs. PMID- 20013814 TI - Melanin-concentrating hormone-1 receptor binding activity of pheophorbides isolated from Morus alba leaves. AB - The time-resolved fluorescence technique based on melanin-concentrating hormone (MCH) receptor subtype-1 (MCH-1 receptor) binding assay was adopted to carry out a bioassay-guided fractionation of the methanol extract of Morus alba leaves. This fractionation and purification led to the isolation of two compounds identified as pheophorbide a methyl ester and 13(2)(S)-hydroxypheophorbide a methyl ester. These active pheophorbides exhibited potent inhibitory activity in binding of europium-labeled MCH to the human recombinant MCH-1 receptor (IC(50) value; 4.03 and 0.33 microM, respectively). Besides binding activity, the pheophorbides inhibited MCH-mediated extracellular signal-regulated kinase (ERK) phosphorylation in Chinese hamster ovary cells expressing human MCH-1 receptor. These results suggest that pheophorbide a methyl ester and 13(2)(S) hydroxypheophorbide a methyl ester act as modulators of MCH-1 receptor and MCH mediated ERK signaling. PMID- 20013815 TI - Sclerocarya birrea (A. Rich) Hochst. ['Marula'] (Anacardiaceae): a review of its phytochemistry, pharmacology and toxicology and its ethnomedicinal uses. AB - A plethora of ethnotherapeutic properties and pharmacological actions have been attributed to Sclerocarya birrea (family: Anacardiaceae). It is one of the most highly valued indigenous trees of southern Africa. Reports in biomedical literature have indicated the presence of medicinally-important chemical constituents in the plant, notably: polyphenols, tannins, coumarins, flavonoids, triterpenoids, phytosterols, and so forth. Pharmacological studies by various groups of investigators have shown that S. birrea possesses antidiarrhoeal, antidiabetic, anti-inflammatory, antimicrobial, antiplasmodial, antihypertensive, anticonvulsant, antinociceptive and antioxidant properties, thus lending pharmacological support to the plant's folkloric, ethnotherapeutic uses in South African traditional medicine. In view of the immense medicinal importance of the plant, this review aimed at compiling all currently available information on S. birrea's chemical constituents, as well as its ethnomedicinal, pharmacological and toxicological properties. PMID- 20013816 TI - Randomized double-blind placebo-controlled trial on the potential modes of action of SheaFlex70 in osteoarthritis. AB - Extracts from the seed of the African shea tree Vitellaria paradoxa C.F. Gaertn have been used traditionally for the treatment of arthritic conditions. However, little is known about the mechanisms by which benefit is conferred. This single site, 15-week randomized, double-blind, parallel, placebo-controlled study examined a range of biomarkers in 89 patients with osteoarthritis of the knees and/or hips to determine potential modes of action of SheaFlex70, a triterpene rich extract of Vitellaria paradoxa. In the group of participants with levels of osteoarthritis biomarkers in the upper quartile at baseline, there were significant decreases in inflammation and cartilage breakdown and trend level decreases in bone remodeling in the SheaFlex70 group versus placebo between commencement and completion of the study. Inflammation marker TNF-alpha fell 23.9% vs 6% (treatment vs placebo), p = 0.041. Cartilage degradation marker CTX II fell 28.7% vs an increase of 17.6% (treatment vs placebo), p = 0.018. This marker also showed significant falls across the entire study group, 10.6% vs an increase of 11.6%, (treatment vs placebo), p = 0.016. Osteocalcin levels fell 9.2%, p = 0.014 (treatment) vs 1.2%, ns (placebo), p = 0.096 (treatment vs placebo). These findings indicate that in patients with the highest levels of osteoarthritis biomarkers, SheaFlex70 demonstrated multiple beneficial activities consistent with slowing the disease process. PMID- 20013817 TI - Antiproliferative and proapoptotic actions of okra pectin on B16F10 melanoma cells. AB - The proliferation and apoptosis of metastatic melanoma cells are often abnormal. We have evaluated the action of a pectic rhamnogalacturonan obtained by hot buffer extraction of okra pods (okra RG-I) on melanoma cell growth and survival in vitro. We added okra RG-I containing an almost pure RG-I carrying very short galactan side chains to 2D (on tissue culture polystyrene, tPS) and 3D (on poly(2 hydroxyethylmethacrylate), polyHEMA) cultures of highly metastatic B16F10 mouse melanoma cells. We then analyzed cell morphology, proliferation index, apoptosis, cell cycle progression and the expression of adhesion molecules. Immunostaining and western blotting were used to assay galectin-3 (Gal-3) protein.Incubation with okra RG-I altered the morphology of B16F10 cells and significantly reduced their proliferation on both tPS and polyHEMA. The cell cycle was arrested in G2/M, and apoptosis was induced, particularly in cells on polyHEMA. The expression of N-cadherin and alpha5 integrin subunit was reduced and that of the multifunctional carbohydrate-binding protein, Gal-3, at the cell membrane increased.These findings suggest that okra RG-I induces apoptosis in melanoma cells by interacting with Gal-3. As these interactions might open the way to new melanoma therapies, the next step will be to determine just how they occur. PMID- 20013818 TI - Mechanisms underlying vascular effect of chronic resveratrol in streptozotocin diabetic rats. AB - In this study, some underlying mechanisms responsible for the beneficial effect of chronic oral administration of resveratrol on aortic reactivity of streptozotocin (STZ)-diabetic rats were investigated. Male diabetic rats received resveratrol (10 mg/kg/day) for 8 weeks, 1 week after diabetes induction. Treatment of diabetic rats with resveratrol produced a hypoglycaemic effect and there were appropriate changes regarding serum lipids. Resveratrol also attenuated the increased malondialdehyde (MDA) content and reduced activity of superoxide dismutase (SOD) in liver and aortic tissues. Maximum contractile response of endothelium-intact aortic rings to KCl and phenylephrine (PE) was significantly lower in resveratrol-treated diabetic rats relative to untreated diabetics. Endothelium removal abolished the significant difference between resveratrol-treated and untreated diabetic groups regarding contractile response to KCl and PE. Meanwhile, endothelium-dependent relaxation to acetylcholine (ACh) was significantly higher in resveratrol-treated diabetic rats as compared to diabetic group and pretreatment with N(omega)-L-arginine methyl ester (L-NAME) and indomethacin (INDO) significantly attenuated these responses. Chronic treatment with resveratrol may prevent diabetes-related changes in vascular reactivity observed in diabetic rats directly and/or indirectly due to its hypoglycaemic and hypolipidaemic effects and attenuation of lipid peroxidation and through endothelial-derived factors. PMID- 20013819 TI - Antitumor and antiplatelet activity of alkaloids from veratrum dahuricum. AB - Ten steroidal alkaloids - cyclopamine, veratramine, jervine, 3, 15 diangyloylgermine, 3-angyloylzygadenine, 3-veratroyl zygadenine, 15 veratroylgermine, germine, veratrosine and pseudojervine - from Veratrum dahuricum, together with the ethanol extract and total alkaloids, were evaluated for their antitumor and antiplatelet activities. Cyclopamine, veratramine and germine significantly inhibited the hedgehog pathway in NIH/3T3 cells. Cyclopamine exerted a potent inhibitory effect against the growth of PANC-1 tumors in mice, with inhibition rates of 40.64%, 44.37%, 46.77% at doses of 5.0, 15.0 and 50.0 mg kg-1, respectively. Veratroylgermine was found to produce the strongest inhibition against the platelet aggregation induced by arachidonic acid, with inhibition rate of 92.0% at 100 microM. PMID- 20013820 TI - Insulin mimetics in Urtica dioica: structural and computational analyses of Urtica dioica extracts. AB - Urtica Dioica (UD) is a plant shown to reduce blood glucose levels upon oral ingestion; however, neither its active component nor its mechanism of action has been identified. One active fraction of this extract, termed UD-1, was separated by molecular sieve column chromatography and purified by high performance liquid chromatography (HPLC). While UD-1 did not stimulate insulin secretion in glucose responsive MIN6 clonal beta-cells, chronic exposure (24 h) significantly enhanced glucose uptake (approximately 1.5-fold) in L6-GLUT4myc myoblast cells. Using HPLC and MALDI-TOF, we further purified the UD-1 fraction into two fractions termed UD 1A and UD-1B. Computational and structural analyses strongly suggested that the antidiabetic component of UD-1 was due to one or more structurally related cyclical peptides that facilitate glucose uptake by forming unique glucose permeable pores. The structure and function of these glucose-conducting pores are discussed herein. PMID- 20013821 TI - Effects of isoflavones equol and genistein on bone quality in a rat osteopenia model. AB - Phytoestrogens might be an alternative medication in prophylaxis and treatment of osteoporosis. In this study, the osteoprotective effects of genistein (GEN) and equol (EQO) were evaluated. After ovariectomy, 44 rats received soy-free food (Control, C) and developed substantial osteoporosis over the course of two months. After that period, the rats were divided into different groups and fed estradiol (E), GEN or EQO for 35 days. To analyze the osteoprotective effects of the tested substances, bone biomechanical properties and histomorphometric changes of the lumbar vertebrae were evaluated. In analyzing the vertebral body compression strength, we found that the EQO (103.8%) and GEN (96.8%) groups reached similar levels relative to the E group, while the C group reached 77.7% of the biomechanical properties of the E group. EQO was significantly superior to C. The histomorphometric evaluation demonstrated an increased number of nodes in EQO- and E-treated rats compared to GEN- and C-treated rats. E led to an improvement of cortical as well as trabecular bone, an advantage that was only partly seen in the other groups. Treatment with phytoestrogens induced improved bone quality. EQO and GEN might be alternatives for hormone replacement therapy, although further studies are needed to elucidate possible side effects. PMID- 20013822 TI - Hypotensive effect of aqueous saffron extract (Crocus sativus L.) and its constituents, safranal and crocin, in normotensive and hypertensive rats. AB - In this study, the effects of saffron (Crocus sativus) stigma aqueous extract and two active constituents, crocin and safranal, were investigated on blood pressure of normotensive and desoxycorticosterone acetate-induced hypertensive rats. Three doses of crocin (50, 100 and 200 mg/kg), safranal (0.25, 0.5 and 1 mg/kg) and the aqueous extract (2.5, 5 and 10 mg/kg) were administered intravenously in different groups of normotensive and hypertensive animals and their effects on mean arterial blood pressure (MABP) and heart rate (HR) were evaluated. The aqueous extract of saffron stigma, safranal and crocin reduced the MABP in normotensive and hypertensive anaesthetized rats in a dose-dependent manner. For example, administrations of 10 mg/kg of aqueous extract, 1 mg/kg of safranal and 200 mg/kg of crocin caused 60 +/- 8.7, 50 +/- 5.2 and 51 +/- 3.8 mmHg reductions in MABP, respectively. It can be concluded that the aqueous extract of saffron stigma has hypotensive properties which appear to be attributable, in part, to the actions of two major constitutes of this plant, crocin and safranal. It seems that safranal is more important than crocin for lowering down blood pressure of rats. PMID- 20013823 TI - (-)-Epigallocatechin-3-gallate increases cell proliferation and neuroblasts in the subgranular zone of the dentate gyrus in adult mice. AB - Neurogenesis is regulated by several factors such as age, stress and pharmacological agents. We observed the effects of (-)-epigallocatechin-3-gallate (EGCG), a major catechin of green tea, on neurogenesis in mice. The animals were orally administered EGCG for 4 weeks. Brain sections were stained using a marker for cell proliferation (Ki67 and BrdU) and neuroblasts (doublecortin, DCX). In all groups, Ki67, BrdU and DCX immunoreaction were observed in the subgranular zone of the dentate gyrus. Oral administration of EGCG significantly increased the number of Ki67-, BrdU- and DCX-immunoreactive cells as well as BrdU/DCX colabled cells in the subgranular zone when compared to those in the vehicle treated group. These results indicate that oral administration of EGCG can enhance cell proliferation and increase the number of neuroblasts in mice hippocampal dentate gyrus. PMID- 20013824 TI - Rapid prototyped PGA/PLA scaffolds in the reconstruction of mandibular condyle bone defects. AB - BACKGROUND: Craniomaxillofacial bone defects are currently reconstructed by using computer-aided design and manufacturing (CAD/CAM) processes. We have developed a novel digital medical support system that enables us to custom-make scaffolds to repair craniomaxillofacial bone defects using three-dimensional computed tomographic (CT) images and a rapid-prototyping method. METHODS: We created positive molds using CT data, CAD/CAM and a rapid prototyping method using 3D printing. Custom-made poly (glycolic acid) (PGA) and polymers poly (lactic acid) (PLA) scaffolds were prefabricated by a positive-negative mold interchange technique. A laser scanning system was used to evaluate the accuracy of the PGA/PLA scaffold. Bone marrow stem cells were incubated with the scaffold to assess biocompatibility. RESULTS: The mean error was <0.3 mm and confidence was >or=95% when the error was <1 mm. Results from in vitro cell culture demonstrated that the PGA/PLA scaffold had excellent cellular compatibility. CONCLUSIONS: This pilot study suggests that custom-made PGA/PLA scaffolds infiltrated with bone marrow stem cells may be effective for future treatment of craniomaxillofacial bone injuries. PMID- 20013825 TI - A medical tactile sensing instrument for detecting embedded objects, with specific application for breast examination. AB - BACKGROUND: In this paper, having considered the tactile sensing and palpation of a physician in order to detect abnormal masses in the breast, we simplified and then modelled the tissue containing a mass and used contact elements to analyse the tactile sensor function. METHODS: By using the finite element method, the effects of the mass existence appeared on the surface of the tissue. This was due to exerting mechanical load on the modelled tissue surface. Following this, a tactile sensing instrument called the 'tactile tumour detector' (TTD) was designed and constructed. This device is able to detect abnormal objects in the simulated models by making contact with model surfaces. In order to perform a series of precise experiments, a robot that could hold the tactile probe was used. The velocity of the linear movement of the probe is low enough to ensure that the tissue behaves in the linear elastic range, so that dynamic effects can be neglected. RESULTS AND DISCUSSION: The maximum value of stresses was chosen as the comparison criterion. The variation of this criterion vs. the mass parameter changes was investigated and good agreements between numerical and experimental results were obtained. Moreover, the sensitivity and specificity of TTD and clinical breast examination (CBE) in the detection of breast masses, in comparison to sonography as the 'gold standard', were calculated by performing clinical trials on 55 cases. PMID- 20013826 TI - Fhl2 interacts with Foxk1 and corepresses Foxo4 activity in myogenic progenitors. AB - Adult skeletal muscle has a remarkable regenerative capacity because of a myogenic progenitor cell population. Using a gene disruption strategy, we determined that Foxk1 regulates myogenic progenitor cell activation and muscle regeneration. In this study, we undertook a yeast two hybrid screen to identify Foxk1 interacting proteins. We identified the LIM-only protein, Fhl2, as a Foxk1 interacting protein. Using transcriptional assays, we observed that Fhl2, in a dose-dependent fashion, promotes Foxk1 transcriptional repression of Foxo4 activity. Using histochemical and immunohistochemical assays, we further established that Fhl2 is expressed in the myogenic progenitor cell population. Fhl2 knockdown results in cell cycle arrest, and mice lacking Fhl2 have perturbed skeletal muscle regeneration. Collectively, these studies define a Fhl2-Foxk1 cascade that regulates the myogenic progenitor cell activity in adult skeletal muscle and enhances our understanding of muscle regeneration. PMID- 20013827 TI - Toward engineering a human neoendothelium with circulating progenitor cells. AB - Tissue-engineered vascular grafts may one day provide a solution to many of the limitations associated with using synthetic vascular grafts. However, identifying a suitable cell source and polymer scaffold to recreate the properties of a native blood vessel remains a challenge. In this work, we assess the feasibility of using endothelial progenitor cells (EPCs) found in circulating blood to generate a functional endothelium on poly(1,8-octanediol-co-citrate) (POC), a biodegradable elastomeric polyester. EPCs were isolated from human blood and biochemically differentiated into endothelial-like cells (HE-like) in vitro. The differentiated cell phenotype and function was confirmed by the appearance of the characteristic endothelial cell (EC) cobblestone morphology and positive staining for EC markers, von Willebrand factor, vascular endothelial cadherin, flk-1, and CD31. In addition, HE-like cells cultured on POC express endothelial nitric oxide synthase at levels comparable to aortic ECs. Furthermore, as with mature endothelial cells, HE-like cell populations show negligible expression of tissue factor. Similarly, HE-like cells produce and secrete prostacyclin and tissue plasminogen activator at levels comparable to venous and aortic ECs. When compared to fibroblast cells, HE-like cells cultured on POC show a decrease in the rate of plasma and whole-blood clot formation as well as a decrease in platelet adhesion. Finally, the data show that HE-like cells can withstand physiological shear stress of 10 dynes/cm(2) when cultured on POC-modified expanded poly(tetrafluoroethylene) vascular grafts. Collectively, these data are the foundation for future clinical studies in the creation of an autologous endothelial cell-seeded vascular graft. PMID- 20013828 TI - Nanobubbles from gas-generating polymeric nanoparticles: ultrasound imaging of living subjects. PMID- 20013829 TI - Reduction/dissolution of a beta-MnOOH nanophase in the ferritin cavity to yield a highly sensitive, biologically compatible magnetic resonance imaging agent. PMID- 20013830 TI - Synthesis of Fe(3)O(4)@SiO(2)@PMMA core-shell-shell magnetic microspheres for highly efficient enrichment of peptides and proteins for MALDI-ToF MS analysis. PMID- 20013831 TI - Self-assembly of rodlike bio-nanoparticles in capillary tubes. PMID- 20013832 TI - Cyclization of TEMPO radicals bound to metalladithiolene induced by SOMO-HOMO energy-level conversion. PMID- 20013834 TI - Metallasilatranes: palladium(II) and platinum(II) as lone-pair donors to silicon(IV). PMID- 20013833 TI - Palladium-catalyzed coupling reactions of thioimidate N-oxides: access to alpha alkenyl- and alpha-aryl-functionalized cyclic nitrones. PMID- 20013835 TI - A green route to well-defined high-molecular-weight (Co)polymers using ARGET ATRP with alkyl pseudohalides and copper catalysis. PMID- 20013836 TI - Porous capsules {(M)M(5)}(12)Fe(III) (30) (M=Mo(VI), W(VI)): sphere surface supramolecular chemistry with 20 ammonium ions, related solution properties, and tuning of magnetic exchange interactions. PMID- 20013838 TI - Graft vascularization is a critical rate-limiting step in skeletal stem cell mediated posterolateral spinal fusion. AB - The ability of skeletal stem cells (SSCs) to direct spinal fusion (SF) upon transplantation in conjunction with osteoconductive biomaterials was investigated in a rabbit model. When tested in a mouse heterotopic transplantation assay, rabbit SSCs and Pro-Osteon 500R was osteoconductive and supported osteogenesis. When used in a SF model, the same constructs induced bone formation in periapophyseal regions (PARs). In this respect, they proved to be superior to grafts of cell-free carrier or total uncultured bone marrow-carrier constructs, used as controls. However, interapophyseal regions (IARs) remained devoid of new bone, such that true bony bridging of adjacent transverse apophyses (true SF) could not be achieved. Interestingly, this could not be predicted from high resolution radiography. A systematic histological survey of the entire graft harvested at 6 months was essential for proper assessment of the transplantation procedure outcome. Immunohistochemical analysis of microvessel density revealed that IARs remained undervascularized, as compared to PARs, suggesting that differential vascularization could account for the absence or presence of new bone formation in the same regions. SF is an extreme model of stem cell-directed bone regeneration, requiring a combination of orthotopic (PAR) and heterotopic (IAR) bone formation. Our data show that, in this setting, graft size can be critical with respect to the necessary neovascularization, a crucial variable independent of proper osteogenic and osteoconductive competence of the cells and materials employed. Furthermore, stringent histological studies are mandatory for proper assessment of outcomes in SF studies, in which the use of mineralized materials can make radiographic assessment misleading. PMID- 20013839 TI - Pyramidal eminence and subpyramidal space: an endoscopic anatomical study. AB - OBJECTIVES/HYPOTHESIS: To describe retrotympanic endoscopic anatomy, especially the pyramidal eminence and contiguous spaces. STUDY DESIGN: This was an anatomical study on a prospective case series. METHODS: The anatomy of the retrotympanum was studied by endoscopy in nine patients affected by cholesteatoma who underwent tympanomastoid surgery and in six temporal bone dissections. RESULTS: Pneumatization of the sinus tympani and posterior tympanic sinus or both, noted in 12 ears out of 15, may give rise to a recess beneath the pyramidal eminence, which we have called the subpyramidal space. This space can manifest with a variable degree of depth, shape, or extent depending on the shape and dimensions of the pyramidal eminence. CONCLUSIONS: Endoscopic exploration of the middle ear may guarantee a very good exposure of retrotympanic structures, allowing detailed anatomical descriptions of hidden areas. Improvement in our knowledge of the anatomy may decrease the possibility of residual disease during cholesteatoma surgery. PMID- 20013841 TI - The shelf life of antimicrobial ear drops. AB - OBJECTIVES/HYPOTHESIS: The advice given to patients regarding the shelf life of antimicrobial ear drops is based on little or no evidence. We aimed to determine appropriate recommendations for the shelf life of common antibiotic-containing topical otic solutions. STUDY DESIGN: Prospective experimental design. METHODS: Ear drops containing gentamicin and ciprofloxacin were analyzed. Their effectiveness was assessed on agar plates inoculated with both Staphylococcus aureus and Pseudomonas aeruginosa. Bottles were also assessed for contamination once opened. In addition, drops used for 1 week on patients with culture-positive otorrhea were analyzed for contamination of both the bottle teat and the drop solution. RESULTS: Neither antibiotic showed deterioration in effectiveness against either microorganism over a 4-month period, as assessed by the size of inhibitory zones on inoculated agar plates. Of drops returned after use on infected patients, none of the cultures from the teats of the bottle grew any organisms. The drop solution itself also showed no contamination after having been opened for 4 months and used by a patient. CONCLUSIONS: There appears to be no evidence that antimicrobial ear drops containing either gentamicin or ciprofloxacin lose their efficacy against either S. aureus or P. aeruginosa over at least 4 months from opening, or that the bottle contents become contaminated after use in patients with culture-positive otorrhea. It is likely these findings apply to other antimicrobial drops and for other organisms. This allows us to advise patients to keep unfinished drops for potential future use with beneficial economic implications. PMID- 20013840 TI - Outcomes of salvage surgery with free flap reconstruction for recurrent oral and oropharyngeal cancer. AB - OBJECTIVES/HYPOTHESIS: To evaluate outcomes of salvage surgery with free flap reconstruction for recurrent squamous cell carcinoma of the oropharynx and oral cavity with increased use of chemoradiotherapy. STUDY DESIGN: Retrospective patient review. METHODS: All patients undergoing salvage surgery with free flap reconstruction for oropharynx (n = 36) and oral cavity (n = 36) squamous cell carcinomas between January 2001 and January 2008 were obtained. Mean follow-up was 14 months. Previous chemoradiotherapy was used in 40% and radiotherapy alone in 60%. RESULTS: Complications were more frequent in oropharynx than oral cavity tumors (36% and 14%, respectively; P = .05) requiring more secondary procedures (15 for oropharynx vs. six for oral cavity). Few patients returned to a normal diet (8%), and a majority retained an enterogastric feeding tube (56%). Median survival overall following salvage surgery was 44.8 months for oral cavity and 53.8 months for oropharynx head and neck squamous cell carcinoma. Overall estimated 1-, 2-, and 5-year observed survivals were 98%, 77.2%, and 43.7%, respectively. Twelve patients had a disease-free interval of <6 months, 92% of whom died of disease. Of 17 patients with disease at the primary site and involved regional lymph nodes, 94% died of disease. CONCLUSIONS: Salvage surgery with free flap reconstruction for recurrent oral and oropharyngeal tumors after chemoradiotherapy has acceptable morbidity and similar cure rates as salvage following radiotherapy without chemotherapy. Concurrent nodal recurrence and short disease-free interval are associated with reduced cure rates. A significant proportion will require enterogastric feeding and few will tolerate a normal diet. PMID- 20013842 TI - Modified laser-assisted stapedotomy. PMID- 20013843 TI - Hedgehog signaling: endocrine gland development and function. AB - The role of hedgehog signaling is analyzed in relation to the developing endocrine glands: pituitary, ovary, testis, adrenal cortex, pancreas, prostate, and epiphyseal growth. Experimental and pathological correlates of these organs are also discussed. The second section addresses a number of topics. First, the pituitary gland, no matter how hypoplastic, is present in most cases of human holoprosencephaly, unlike animals in which it is always said to be absent. The difference appears to be that animal mutations and teratogenic models involve both copies of the gene in question, whereas in humans the condition is most commonly heterozygous. Second, tests of endocrine function are not reported with great frequency, and an early demise in severe cases of holoprosencephaly accounts for this trend. Reported tests of endocrine function are reviewed. Third, diabetes insipidus has been recorded in a number of cases of holoprosencephaly. Its frequency is unknown because it could be masked by adrenal insufficiency in some cases and may not be recognized in others. Because of the abnormal hypothalamic-infundibular region in holoprosencephaly, diabetes insipidus could be caused by a defect in the supra-optic or paraventricular hypothalamic nuclei or in release of ADH via the infundibulum and posterior pituitary. PMID- 20013844 TI - Role of the otolaryngologist in the treatment of struma ovarii. AB - Struma ovarii, a rare germ cell tumor of the ovary composed of >50% thyroid tissue, is traditionally managed by gynecologic surgeons. Although struma ovarii is typically cured by simple excision, otolaryngology-head and neck surgeons may play a critical role in caring for these patients. In this article, we present two cases of struma ovarii and discuss the role of the otolaryngologist. PMID- 20013845 TI - In reference to Squamous cell carcinoma associated with intralesional injection of cidofovir for recurrent respiratory papillomatosis. PMID- 20013846 TI - Rhinovirus infection-induced alteration of tight junction and adherens junction components in human nasal epithelial cells. AB - OBJECTIVES/HYPOTHESIS: Manifestations of rhinovirus (RV) infections include mucus overproduction, increased vascular permeability, and secondary bacterial infection. These effects may reflect disrupted epithelial barrier functions, which are mainly regulated by intercellular junctions, referred to as tight junctions (TJs) and adherens junctions (AJs). The objective of this study was to investigate changes in the components of TJs (ZO-1, occluding, and claudin-1) and AJs (E-cadherin) after RV infection in cultured nasal epithelial cells. METHODS: Primary human nasal epithelial cells grown at an air-liquid interface were infected apically with RV. RV-induced changes in the expression of epithelial TJ and AJ proteins were determined using real-time reverse transcriptase-polymerase chain reaction, confocal microscopy, and Western blot analyses. Functional changes in the integrity of junctional proteins were assessed by measuring transepithelial resistance (TER) using a voltmeter. RESULTS: RV infection decreased mRNA levels of ZO-1, occludin, claudin-1, and E-cadherin to 64.2%, 51.8%, 56.2%, and 56.3%, respectively, of those in controls (P < .05). Decreases in ZO-1, occludin, claudin-1, and E-cadherin protein levels in RV-infected cells were evident in immunofluorescent confocal microscopic images. Expression levels of these proteins were also lower in the RV-infected group in Western blot analyses. RV infection reduced the mean TER from 143.1 Omega/cm(2) (controls) to 122.6 Omega/cm(2). CONCLUSIONS: RV infection decreased the expression of TJ and AJ components and reduced TER in primary cultured human nasal epithelial cells, indicating that RV infection may exert a harmful effect on nasal epithelial barrier function. PMID- 20013847 TI - Impact of thyroidectomy without laryngeal nerve injury on vocal quality characteristics: an objective multiparameter approach. AB - OBJECTIVES/HYPOTHESIS: The main purpose of this study was to determine the impact of thyroidectomy on the subjective and objective vocal quality using the Dysphonia Severity Index. It was hypothesized that objective measures of vocal function and other vocal characteristics would change (temporarily or permanently) from the presurgical to the postsurgical conditions, even with the entire preservation of the laryngeal nerve, due to the surgical approach and other influencing factors. STUDY DESIGN: Prospective study in which 44 subjects were studied before (1 week) and three times post-thyroidectomy. METHODS: Subjective (auditory perceptual evaluation and videolaryngostroboscopy) and objective (aerodynamic, vocal range, acoustic, and Dysphonia Severity Index measurements) assessment techniques were used. RESULTS: Paired samples t test indicated a significant decrease of the highest frequency, the highest intensity, the fundamental frequency, and the Dysphonia Severity Index in the first postoperative condition. When a repeated measures analysis of variance was performed with the pre- and all the postoperative moments of evaluation taken into account, no significant difference was noted for any of the objective voice characteristics. CONCLUSIONS: After thyroidectomy subjects had a normal perceptual and objective vocal quality corresponding with a Dysphonia Severity Index of 66%, and there is no permanent change of the vocal performance. Moreover, there is no psychosocial handicapping effect of the vocal quality, but immediately post-thyroidectomy there are more vocal complaints. To what extent thyroidectomy causes (temporary or permanent) alterations of the singing voice in elite vocal performers is subject to further research. PMID- 20013848 TI - Modulation of vocal fold scar fibroblasts by adipose-derived stem/stromal cells. AB - OBJECTIVES/HYPOTHESIS: To explore whether adipose-derived stem/stromal cells (ASCs) have therapeutic potential for treating scarred superficial lamina propria through the effects of secreted hepatocyte growth factor (HGF) on scar fibroblasts. STUDY DESIGN: In vitro study using coculture system. METHODS: Scar fibroblasts (SFs) were isolated from ferret vocal folds electrocauterized 2 weeks previously. ASCs were isolated from ferret lipoaspirated subcutaneous abdominal fat. For coculture experiments, the two cell types were combined in Transwell plates for 6 days, followed by 1 or 3 days of monoculture after removing the upper chamber. Assays were then performed on cells and media from the bottom chamber. We measured: 1) the production of hyaluronic acid (HA), collagen and HGF via enzyme-linked immunosorbent assays, 2) the expression of alpha-smooth muscle actin (alpha-SMA), 3) cell proliferation, and 4) apoptosis of SFs (2, 3, and 4 via flow cytometry). Other experiments examined the effects of HGF on SFs and the effects of HGF neutralization in the coculture system. RESULTS: Coculture led to significant decreases in SF collagen production (P < .05), cell proliferation (P < .05), and alpha-SMA expression (P < .05), whereas HA production increased (P < .05). Coculture also increased HGF secretion from ASCs (P < .05). Neutralization of HGF abolished the inhibitory effects of ASCs on SF collagen synthesis (P < .05). CONCLUSIONS: ASCs influence SFs to adopt a less fibrotic profile. It appears that HGF is at least one of the soluble factors responsible for this effect. Implanted ASCs could potentially ameliorate vocal fold scar by acting as long-term, intrinsic sources of HGF. PMID- 20013849 TI - Diagnosis and management of pediatric cervicofacial venous malformations: retrospective review from a vascular anomalies center. AB - OBJECTIVES/HYPOTHESIS: Venous malformations are rare congenital aberrations of vein development frequently presenting in the head and neck. Without treatment, venous malformations cause progressive disfigurement, dysfunction, and bleeding. In this study, we analyzed a cohort of pediatric patients with cervicofacial venous malformations (CFVM), with the goal of developing a treatment algorithm for these complex lesions. STUDY DESIGN: Eleven-year retrospective chart review. The setting was a vascular anomalies center at a pediatric tertiary hospital. Nineteen patients (10 male, 9 female), aged 11 months to 17 years, presented with CFVM. METHODS: Patient charts were reviewed for demographics, signs and symptoms, timing of first and subsequent interventions, total number and type of interventions performed, and procedural complications. A family questionnaire supplemented outcome measures by determining the perception of disease control. RESULTS: Presenting symptoms for CFVM include growth (100%), disfigurement (63%), pain (58%), respiratory compromise (42%), and dysphagia (32%). A mean of 6.7 interventions were performed per patient (median, 6; range, 2-12), requiring a mean of 0.8 excisions, 4.6 laser treatments, and 1.3 sclerotherapy injections. Average age at first procedure was 8.5 years. Time between treatments averaged 8.9 months. Four complications occurred in 127 procedures (3.1%). Questionnaire responses indicated subjective improvement following therapy. A management algorithm could be developed from therapeutic outcomes. CONCLUSIONS: Treatment of CFVM can be safely and successfully performed with a combination of laser therapy, sclerotherapy, and surgical excision. A treatment algorithm involving multiple procedures during childhood can lead to successful management of CFVM. PMID- 20013850 TI - Transoral laser ventricular fold resection and thyroarytenoid myoneurectomy for adductor spasmodic dysphonia: long-term outcome. AB - OBJECTIVES: A transoral approach to laser myoneurectomy of the thyroarytenoid muscles was recently reported as an effective technique for treatment of adductor spasmodic dysphonia (ASD). The short-term results were encouraging. In this study, a long-term surgical outcome of this technique is investigated. STUDY DESIGN: A prospective clinical series. METHODS: Fifty-two patients with ASD underwent transoral laryngomicrosurgery with a CO(2) laser to resect the ventricular folds followed by myoneurectomy of the thyroarytenoid muscles. The nerve fibers of recurrent laryngeal nerve terminating at the thyroarytenoid muscle, which were frequently found branching deeply among the posterior belly of this muscle, were vaporized. Care was taken not to damage the vocal ligaments, lateral cricoarytenoid muscles, or arytenoid cartilages. Pre- and postoperative subjective voice assessments, and acoustic and aerodynamic measurements, were performed and statistically analyzed. RESULTS: Twenty-nine of the 52 patients who were followed up for more than 12 months (range, 12-63 months; mean, 31 months) were studied. Moderate and marked vocal improvement was achieved in 90% (26/29) of the patients. Three patients had 30% improvement, which was reported as unsuccessful. Eight of the 26 patients who were satisfied with their postoperative voice still had a mild strain during phonation. Of the eight patients, seven achieved normal or nearly normal voice quality after revision laser surgery. No significant vocal deficit or paralysis was observed in any patient. CONCLUSIONS: After long-term follow-up of 31 months on average, approximately 90% of the ASD patients obtained moderate to marked improvement of vocal performance after transoral laser surgery. The long-lasting outcome is encouraging. PMID- 20013851 TI - Virtual reality: a new paranasal sinus surgery simulator. AB - OBJECTIVES/HYPOTHESIS: Virtual surgical training systems are of growing value. Current prototypes for endonasal sinus surgery simulation are very expensive or lack running stability. No reliable system is available to a notable number of users yet. The purpose of this work was to develop a dependable simulator running on standard PC hardware including a detailed anatomic model, realistic tools and handling, stereoscopic view, and force feedback. STUDY DESIGN: Descriptive. METHODS: A three-dimensional voxel model was created based on a high-resolution computed tomography study of a human skull, from which the bony structures were segmented. The mucosa and organs at risk were added manually. The model may be manipulated with virtual surgical tools controlled with a low-cost haptic device, which is also used to adjust microscopic or endoscopic views. Visualization, haptic rendering, and tissue removal are represented with subvoxel resolution. RESULTS: The handling of the model is convincing. The haptic device provides a realistic feeling regarding the interaction between tool tip and anatomy. Three dimensional orientation and the look and feel of virtual surgical interventions get close to reality. CONCLUSIONS: The newly developed system is a stable, fully operational simulator for sinus surgery based on standard PC hardware. Besides the limitations of a low-cost haptic device, the presented system is highly realistic regarding anatomy, visualization, manipulation, and the appearance of the tools. It is mainly intended for gaining surgical anatomy knowledge and for training navigation in a complex anatomical environment. Learning effects, including motor skills, have yet to be quantified. PMID- 20013853 TI - Understanding communication during hospitalist service changes: a mixed methods study. AB - BACKGROUND: Little data exist to inform hospitalist communication during service changes. OBJECTIVE: To characterize hospitalist handoffs during service changes. DESIGN: Serial survey study. SETTING: Single academic medical center. MEASUREMENTS: From May to December 2007, 60 service changes among 17 hospitalists on a nonteaching service were targeted for evaluation using an anonymous 18-item survey that was completed by hospitalists within 48 hours of assuming care for patients. Survey items assessed completeness of handoff communication, certainty of patient care plans, missed information, time spent recovering information, and near misses/adverse events due to incomplete handoffs. The association between completeness of communication and handoff outcomes was examined. Narrative comments were analyzed qualitatively. RESULTS: Ninety-three percent (56/60) of surveys were returned. All 17 hospitalists participated. Thirteen percent of respondents reported incomplete handoffs and 18% were uncertain of care plan on transition day. At least 1 near miss, attributable to incomplete communication was reported by 16%. Hospitalists who reported incomplete handoffs were more likely to report uncertainty about patient care plans on the transition day (71% incomplete vs. 10% complete, P < 0.01), discovery of missing information (71% incomplete vs. 24% complete, P = 0.01), near misses/adverse events (57% incomplete vs. 10% complete, P < 0.01), and more time resolving issues arising from missed information (71% incomplete vs. 22% complete, P < 0.01). Qualitative comments suggest the need for a more systematic, focused, team-based, and patient centered handoff model. CONCLUSIONS: Incomplete handoffs during service changes are associated with uncertainty and potential patient harm. Suggestions to improve the completeness of hospitalist service change communications are offered. PMID- 20013852 TI - California hospital leaders' views of hospitalists: meeting needs of the present and future. AB - BACKGROUND: Hospital medicine has grown rapidly, but hospital leaders' perceptions of current and future drivers of hospitalist growth are unknown. OBJECTIVE: : To determine hospital executives' perceptions of factors leading to hospitalist implementation and their vision for hospitalists' work roles. SETTING: Nonfederal, acute care hospitals in California. PARTICIPANTS: California hospital leaders (eg, chief executive officers). INTERVENTION: Cross-sectional survey from 2006 to 2007. MEASUREMENTS: We asked California hospital leaders whether their hospitals had a hospitalist service and the prospects for growth. In addition, we examined factors responsible for implementation, scope of hospitalists' practices, and need for additional certification as perceived by hospital leaders. RESULTS: We received surveys from 179 of 334 hospitals (response rate of 54%). Of the 64% of respondents that reported the use of hospitalists, none intended to decrease the size of their hospitalist group, and 57% expected growth over 2 years. The most common reasons for implementing a hospitalist program were to care for uncovered patients (68%) and improve cost/length of stay (63%). Respondents also indicated that demand from other physicians was an important factor. Leaders reported that hospitalists provide a wide range of services, with a majority involved in quality improvement projects (72%) and medical comanagement of surgical patients (66%). Most leaders favor additional certification for hospitalists. CONCLUSIONS: There is widespread adoption of hospitalists in California hospitals, with an expectation of continued growth. The drivers of the field's growth are evolving and dynamic. In particular, attentiveness to quality performance and demand from other physicians are increasingly important reasons for implementation. PMID- 20013854 TI - Pharmacokinetics and biliary excretion of mitoxantrone in rats. AB - The objective of this investigation was to compare the observed biliary clearance (CL(b)) and % of dose excreted in the bile (PD(b)) of mitoxantrone with the predicted values obtained from quantitative structure pharmacokinetic relationship (QSPKR) models. Blood and bile samples were collected from bile duct cannulated rats after an intravenous bolus dose of 0.5 or 2 mg/kg mitoxantrone, and the concentrations were measured by HPLC. Mitoxantrone plasma concentrations exhibited a tri-exponential profile with systemic clearance of 118 +/- 6.8 mL/min/kg. After dosing, 6.08 +/- 2.32% and 5.69 +/- 0.59% of the dose were excreted into bile in unchanged form after a 3-h collection. CL(b) was 7.20 +/- 4.54 and 7.46 +/- 0.62 mL/min/kg after the two doses. With the co-administration of 10 mg/kg GF-120918, a P-glycoprotein and BCRP inhibitor, PD(b) was reduced to 0.69 +/- 0.07%, suggesting that BCRP or P-glycoprotein may play an important role in the biliary elimination of mitoxantrone. Using QSPKR models developed for the biliary excretion of cations/neutral compounds in rats, CL(b) and PD(b) of mitoxantrone were predicted as 5.18 mL/min/kg and 7.21%, respectively, suggesting that the models could be used to predict the biliary excretion of mitoxantrone. PMID- 20013856 TI - Role of hospitalists in an offsite alternate care center (ACC) for pandemic flu. AB - Recent concerns about an influenza pandemic have highlighted the need to plan for offsite Alternate Care Centers (ACCs). The likelihood of a successful response to patient surges will depend on the local health systems' ability to prepare well in advance of an influenza pandemic. Our health system has worked closely with our state's medical biodefense network to plan the establishment of an ACC for an influenza pandemic. As hospitalists have expanded their roles in their local health systems, they are poised to play a major role in planning for the next influenza pandemic. Hospitalists should work with their health system's administration in developing an ACC plan. PMID- 20013858 TI - Periprocedural antithrombotic management: a review of the literature and practical approach for the hospitalist physician. AB - Many patients who are on long-term antithrombotic therapy (e.g. warfarin and/or antiplatelet agents) must be assessed for temporary discontinuation for a procedure or surgery, making this a salient topic for the hospitalist physician. Discontinuation of antithromhotic therapy can place patients at increased risk of thromboembolic complications while continuing antithrombotic therapy can increase procedure-related bleeding risk. Bridging anticoagulation with heparin or low molecular weight heparins is often used in the periprocedural period, but a great deal of uncertainty exists about how and when to use bridging anticoagulation. Because there is very little Level 1 evidence to define optimal care, both clinical practice and expert consensus guideline opinions vary. For the hospitalist, it is of critical importance to understand the available data, controversies, and management options in order to approach patient care rationally. This review provides a step-wise literature-based discussion addressing the following four questions: (1) What is the optimal management of antiplatelet therapy in the periprocedural period? (2) Are there very low bleeding risk procedures that do not require interruption of oral anticoagulation? (3) Are there low thromboembolic risk populations who do not require periprocedural bridging? (4) How do you manage patients who must discontinue anti-coagulants but are at an increased thrombotic risk? PMID- 20013859 TI - U.S. physician satisfaction: a systematic review. AB - INTRODUCTION: There is concern in the US about the burden and potential ramifications of dissatisfaction among physicians. The purpose of this article is to systematically review the literature on US physician satisfaction. METHODS: A MEDLINE search with the medical subject headings (MeSH) phrases: (physicians OR physician's role OR physician's women) AND (job satisfaction OR career satisfaction OR burnout), limited to humans and abstracts, with 1157 abstracts reviewed. After exclusions by 2 independent reviewers, 97 articles were included. Physician type sampled, sample size/response rate, satisfaction type, and satisfaction results were extracted for each study. Satisfaction trends were extracted from those studies with longitudinal or repeated cross sectional design. Variables associated with satisfaction were extracted from those studies that included multivariate analyses. RESULTS: Physician satisfaction was relatively stable, with small decreases primarily among primary care physicians (PCPs). The major pertinent mediating factors of satisfaction for hospitalists include both physician factors (age and specialty), and job factors (job demands, job control, collegial support, income, and incentives). CONCLUSIONS: The majority of factors associated with satisfaction are modifiable. Tangible recommendations for measuring and diminishing dissatisfaction are given. PMID- 20013860 TI - Flushing out the diagnosis. PMID- 20013861 TI - A new perspective. PMID- 20013862 TI - Cognitive interview techniques reveal specific behaviors and issues that could affect patient satisfaction relative to hospitalists. AB - BACKGROUND: There is a trend toward patient-centered care as a means of improving patient satisfaction. The Centers for Medicare and Medicaid Services (CMS) have made this concept more significant with plans to link reimbursement to patient satisfaction measures such as the Hospital Consumer Assessment of Healthcare Providers and Systems survey (HCAHPS). OBJECTIVES: To generate hypotheses for reasons underlying diminished HCAHPS patient satisfaction survey ratings, with reference to hospitalists. DESIGN, SETTING, PATIENTS: Observational study conducted using a cognitive interview (CI) technique in a 180-bed community hospital on adult medical, surgical, and critical care inpatients. MEASUREMENTS: Mixed qualitative and quantitative study using both standard responses and open ended responses. The standard responses were compiled into raw numbers and percentages and the qualitative responses were evaluated for common themes and other useful information. RESULTS: Notable factors that may affect satisfaction of patients include ability to have all of their questions answered, incomplete discussion of medication side effects, and failure of physicians to listen and form personal connections with them. CONCLUSION: Cognitive interview techniques can be used to provide additional detail regarding patient satisfaction beyond that provided by standard surveys. PMID- 20013863 TI - Inpatient glucose control: a glycemic survey of 126 U.S. hospitals. AB - BACKGROUND: Despite increased awareness of the value of treating inpatient hyperglycemia, little is known about glucose control in U.S. hospitals. METHODS: The Remote Automated Laboratory System-Plus (RALS-Plus Medical Automation Systems, Charlottesville, VA) was used to extract inpatient point-of-care bedside glucose (POC-BG) tests from 126 hospitals for the period January to December 2007. Patient-day-weighted mean POC-BG and hypoglycemia/hyperglycemia rates were calculated for intensive care unit (ICU) and non-ICU areas. The relationship of POC-BG levels with hospital characteristics was determined. RESULTS: A total of 12,559,305 POC-BG measurements were analyzed: 2,935,167 from the ICU and 9,624,138 from the non-ICU. Patient-day-weighted mean POC-BG was 165 mg/dL for ICU and 166 mg/dL for non-ICU. Hospital hyperglycemia (>180 mg/dL) prevalence was 46.0% for ICU and 31.7% for non-ICU. Hospital hypoglycemia (<70 mg/dL) prevalence was low at 10.1% for ICU and 3.5% for non-ICU. For ICU and non-ICU there was a significant relationship between number of beds and patient-day-weighted mean POC BG levels, with larger hospitals (> or = 400 beds) having lower patient-day weighted mean POC-BG per patient day than smaller hospitals (<200 beds, P < 0.001). Rural hospitals had higher POC-BG levels compared to urban and academic hospitals (P < 0.05), and hospitals in the West had the lowest values. CONCLUSIONS: POC-BG data captured through automated data management software can support hospital efforts to monitor the status of inpatient glycemic control. From these data, hospital hyperglycemia is common, hypoglycemia prevalence is low, and POC-BG levels vary by hospital characteristics. Increased hospital participation in data collection and reporting may facilitate the creation of a national benchmarking process for the development of best practices and improved inpatient hyperglycemia management. PMID- 20013864 TI - Clinical utility of abnormal opacity overlying the vertebral column on lateral chest radiography. AB - BACKGROUND: Chest radiography is an important component of the evaluation of patients with complaints referable to the chest. We sought to investigate the clinical utility of one particular finding on the lateral chest radiograph (CXR), namely, radioopacity obscuring the normal superior to inferior progression of vertebral radiolucency. A review of the literature yielded little published evidence to characterize the clinical utility of this finding to date. METHODS: We retrospectively identified 370 patients from a hospital database who underwent both computed tomography (CT) imaging of the chest and lateral chest radiography within 24 hours. We calculated the sensitivity, specificity, and likelihood ratios (LRs) associated with the presence or absence of an abnormal opacity overlying the vertebral column on lateral chest radiography using CT imaging of the chest as the reference standard. We also estimated interobserver and intraobserver reliability of this finding. RESULTS: Abnormal opacity overlying the vertebral column had a sensitivity of 86.9% (95% confidence interval [CI], 82.5%-90.3%) and specificity of 70.4% (95% CI, 59.7%-79.2%) for relevant CT documented lower lobe and associated structural pathology. The associated summary positive LR (LR+) was 2.9 (95% CI, 2.1-4.1) and summary negative LR (LR-) was 0.19 (95% CI, 0.13-0.26). Kappa statistics were indicative of moderate intraobserver and interobserver agreement. CONCLUSIONS: The presence of abnormal opacity overlying the vertebral column on lateral chest radiography increases the probability of lower lobe and associated structural pathology somewhat. The absence of this finding decreased the probability of such pathology to a greater degree. Thus, this finding is useful in differentiating those patients with pathology from those without. PMID- 20013865 TI - Brugada syndrome unmasked by a mosquito. PMID- 20013866 TI - Gastric outlet obstruction as the presenting manifestation of metastatic lobular breast carcinoma. PMID- 20013867 TI - A life threatening complication of anticoagulation prophylaxis-bilateral adrenal hemorrhage. PMID- 20013868 TI - Ontogeny of robusticity of craniofacial traits in modern humans: a study of South American populations. AB - To date, differences in craniofacial robusticity among modern and fossil humans have been primarily addressed by analyzing adult individuals; thus, the developmental basis of such differentiation remains poorly understood. This article aims to analyze the ontogenetic development of craniofacial robusticity in human populations from South America. Geometric morphometric methods were used to describe cranial traits in lateral view by using landmarks and semilandmarks. We compare the patterns of variation among populations obtained with subadults and adults to determine whether population-specific differences are evident at early postnatal ontogeny, compare ontogenetic allometric trajectories to ascertain whether changes in the ontogeny of shape contribute to the differentiation of adult morphologies, and estimate the amount of size change that occurs during growth along each population-specific trajectory. The results obtained indicate that the pattern of interpopulation variation in shape and size is already established at the age of 5 years, meaning that processes acting early during ontogeny contribute to the adult variation. The ontogenetic allometric trajectories are not parallel among all samples, suggesting the divergence in the size-related shape changes. Finally, the extension of ontogenetic trajectories also seems to contribute to shape variation observed among adults. PMID- 20013869 TI - Metabolomics: will it stay? PMID- 20013871 TI - Potential attractants for detecting and removing invading Gambian giant pouched rats (Cricetomys gambianus). AB - BACKGROUND: Native to Africa, Gambian giant pouched rats (Gambian rats; Cricetomys gambianus Waterh.) are a threatening invasive species on a Florida island, Grassy Key. Gambian giant pouched rats shifted from a domestic pet to invading species after suspected release from a pet breeder. Because of the large size of Gambian rats (weighing up to 2.8 kg), they pose a serious threat to native species (particularly nesting species) and agricultural crops, especially if Gambian rats invade mainland Florida. Also, Gambian rats pose a threat from disease, as they were implicated in a monkeypox outbreak in the midwestern United States in 2003. The United States Department of Agriculture's Wildlife Services has initiated eradication and detection efforts in the Florida Keys, but trapping the sparse population of Gambian rats has proven difficult. RESULTS: Fifteen attractants that could be used in traps for capturing or detecting single or paired Gambian rats were tested. It was found that conspecific scents (i.e. feces and urine) from other Gambian rats were the best treatment for attracting single and paired Gambian rats. Single Gambian rats explored more attractant types than paired Gambian rats. CONCLUSIONS: Effective attractants for use with Gambian rats have been identified, and multiple attractant types should be used to capture or detect the sparse population. It is recommended that mainly urine and feces from Gambian rats be used, but peanut butter, anise, ginger and fatty acid scent could also be useful for attracting the currently small population of Gambian rats on Grassy Key. PMID- 20013872 TI - ADAM 12 may be used to reduce the false positive rate of first trimester combined screening for Down syndrome. AB - BACKGROUND: ADAM12 has been shown to be an efficient maternal serum marker for Down syndrome (DS) in the first trimester; but recent studies, using a second generation assay, have not confirmed these findings. We examined the efficiency of a second generation assay for ADAM12. MATERIALS AND METHODS: ADAM12 concentrations were determined in 28 first trimester DS and 503 control pregnancies using a novel Research Delfia ADAM12 kit. Log10MoM distributions of ADAM12 and correlations with other markers were established. Population performance of screening was estimated by Monte Carlo simulation. RESULTS: ADAM12 was significantly reduced in the first trimester in DS pregnancies with a log10MoM of -0.1621 (equivalent to 0.68 MoM) (p < 0.001). The reduction decreased with advancing gestational age. ADAM12 used with PAPP-A + hCG beta + NT (CUB screening) increased the detection rate (DR) from 86% to 89% for a false positive rate (FPR) of 5%. When used for a fixed DR of 90%, the addition of ADAM12 resulted in a 25% reduction of the FPR. CONCLUSION: ADAM12 is a moderately effective DS marker. It is not a cost-effective addition to CUB screening, but may be used to reduce the FPR in selected high-risk cases. PMID- 20013873 TI - Free leptin index and PAPP-A: a first trimester maternal serum screening test for pre-eclampsia. AB - BACKGROUND: Prophylaxis with low-dose aspirin may reduce the risk of pre eclampsia (PE) if introduced in first trimester. The performance of first trimester maternal serum screening for PE using free leptin index (fLI) and PAPP A, where fLI = leptin/leptin soluble receptor was studied. METHODS: First trimester serum samples from 126 PE pregnancies and 289 control pregnancies were studied. fLI and PAPP-A were converted into gestational age and maternal weight independent log MoM values of PAPP-A and fLI. The screening performance of markers was studied by receiver-operator-characteristics curves. The performance of population screening was estimated by Monte Carlo simulation. RESULTS: fLI was significantly (p < 0.001) elevated [mean log MoM 0.2165 (SD: 0.2604)] compared to controls [mean log MoM -0.0368 (SD: 0.3132)] and PAPP-A was significantly (p < 0.001) reduced [mean log MoM -0.0133 (SD: 0.2661)] compared to controls [mean log MoM 0.0474 (SD: 0.2521)] in PE pregnancies. There was no correlation between fLI and PAPP-A in control or PE pregnancies. Combined fLI and PAPP-A screening for PE had estimated population detection rates of 22% and 35% for false positives rates of 6% and 12%, respectively. CONCLUSION: Combining PAPP-A and fLI improves screening performance for PE compared to single marker screening. PMID- 20013874 TI - A case of Tessier number 7 cleft with severe micrognathia: prenatal sonographic and three-dimensional helical computed tomographic images. PMID- 20013875 TI - Lack of patient knowledge regarding hospital medications. AB - BACKGROUND: Patient involvement in preventing inpatient medication errors is predicated upon patient knowledge of their medications. However, there is little published on the accuracy of patient knowledge or understanding of their hospital medications. OBJECTIVE: To assess hospitalized patients' knowledge of their hospital medications and attitudes towards involvement in the medication safety process while hospitalized. METHODS: A cross-sectional study of 50 adult internal medicine inpatients at the University of Colorado Hospital. Patients completed a list of the hospital medications they believed were prescribed to them and a survey of attitudes toward involvement in the medication safety process. The patient-completed hospital medication list was compared to the hospital medication administration record. RESULTS: Ninety-six percent of study patients omitted at least one prescribed hospital medication. On average, patients omitted 6.8 hospital medications. Forty-four percent of patients believed they were receiving at least one hospital medication that was not actually prescribed. Patients < 65 years old omitted 60% of their as needed (PRN) medications whereas patients > or = 65 years old omitted 88% (P = 0.01). Only 28% reported having seen their hospital medication list, although 81% reported this would improve their satisfaction with hospital care. Ninety percent wanted to review their hospital medication list for accuracy and 94% felt patient review of the hospital medication list had the potential to reduce errors. CONCLUSIONS: Our findings suggest that, in contrast to patient preferences, there are significant deficits in patients' knowledge of hospital medications. These results are a call to reexamine how we educate patients regarding their hospital medications. PMID- 20013876 TI - How much do family physicians involve pregnant women in decisions about prenatal screening for Down syndrome? AB - OBJECTIVE: To assess the extent to which family physicians (FPs) involve women in decisions about prenatal screening for Down syndrome. METHODS: Based on transcripts of consultations between 41 FPs and 128 women, two raters independently assessed clinician's efforts to involve women in decisions about prenatal screening for Down syndrome using the French-language version of OPTION. Descriptive statistics of OPTION scores were calculated. Construct validity was assessed by performing a principal factor analysis and by measuring association with consultation duration and FPs sociodemograhics. Internal consistency was assessed with Cronbach's alpha and inter-rater reliability with the intraclass correlation coefficient. RESULTS: The overall mean OPTION score was low: 19 +/- 7 (range = 0 [no involvement] to 100 [high involvement]). One factor accounted for 80% of the variance. Both internal consistency and inter-rater reliability were very good (Cronbach's alpha = 0.73; ICC = 0.76). OPTION scores were lower for residents than for licensed FPs (17 +/- 5 vs 21 +/- 4; p = 0.02) and were positively associated with duration of consultation (r = 0.56; p < 0.001). CONCLUSION: Based on the French-language version of OPTION, which showed satisfactory psychometric properties, FPs studied put minimal efforts to involve women in decisions about prenatal screening for Down syndrome. PMID- 20013877 TI - Effects of fungicides on in vitro spore germination and mycelial growth of the phytopathogens Leptosphaeria maculans and L. biglobosa (phoma stem canker of oilseed rape). AB - BACKGROUND: Phoma stem canker, caused by the coexisting related fungal pathogens Leptosphaeria maculans (Des.) Ces. & de Not and L. biglobosa Shoemaker & H Brun, is a major disease of winter oilseed rape in the UK. Annually, over 90% of UK crops receive at least one foliar application of fungicide, but little is known about the sensitivity of the more damaging L. maculans and the less damaging L. biglobosa to these fungicides. The effects of flusilazole, tebuconazole and Methyl Benzimidazole Carbamate (MBC) fungicides (benomyl and carbendazim) on the germination of ascospores, conidia and germ tube growth of both species were examined. Isolates collected from different oilseed rape crops in England and Wales were assessed for their mycelial growth on fungicide-amended medium, and ED(50) values were calculated. RESULTS: Leptosphaeria maculans and L. biglobosa differed in their sensitivity to fungicides. Conidial germination of L. maculans was more sensitive to these fungicides than that of L. biglobosa. Isolates of L. maculans had smaller ED(50) values for mycelial growth for all fungicides tested than isolates of L. biglobosa. CONCLUSION: These results suggest that fungicide applications might affect the structure of L. maculans/L. biglobosa populations in UK winter oilseed rape crops. PMID- 20013878 TI - Inter-organizational collaboration projects in the public sector: a balance between integration and demarcation. AB - For several years, the development of the Swedish public sector has been accompanied by a discussion about inter-organizational collaboration, which has been examined in several national experiments. The experience, however, indicates significant difficulties in implementing collaboration in local authorities' regular activities. This article argues that organizing inter-organizational collaboration in projects tends to be counterproductive, since the purpose of this collaboration is to increase the integration of local authorities. This article is based on case studies of three different collaboration projects. Each project is analyzed in relation to the way collaboration is organized within the project and how the relationship to the local authorities' activities is designed. The outcome of these studies shows that while collaboration projects increase integration between the responsible authorities, the integration stays within the projects. This is due to the fact that the projects were designed as units separate from the responsible authorities. As a result, the collaboration that occurs in the projects is not implemented in the local authorities' activities, and the viability of the increased integration of different responsible authorities does not extend beyond the projects. PMID- 20013879 TI - Evaluation of the factor structure and psychometric properties of the Brief Symptom Inventory-18 with homebound older adults. AB - OBJECTIVE: Homebound older adults are at high risk for depression and anxiety. Systematic screening may increase identification of these difficulties and facilitate service usage. The purpose of this study was to investigate the factor structure, internal consistency, and concurrent validity of the Brief Symptom Inventory-18 (BSI-18) for use as a screening instrument for depression and anxiety with homebound older adults and to examine if the BSI-18 could be shortened further and exhibit comparable psychometric properties. METHODS: A sample of 142 older adults receiving in-home aging services completed interviews that included the BSI-18 and the depression and anxiety modules of the structured clinical interview for DSM-IV. RESULTS: Confirmatory factor analysis showed that the theorized three-factor, second-order model of the BSI-18 fit the data well (S B X(2) = 136.17; p = 0.36). The depression and anxiety subscales exhibited high internal consistency (alpha > 0.81), whereas the somatic subscale exhibited lower internal consistency (alpha = 0.69). Receiver operator curve (ROC) analyses suggest that the BSI-18 depression and anxiety subscales were able to predict those with DSM-IV diagnoses (Depression AUC = 0.89 p < 0.001; Anxiety AUC = 0.80, p < 0.001). The ROC results suggested adapting a cut score of T = 50 to achieve optimal sensitivity and specificity. The short three-item depression scale exhibited comparable psychometric properties to the full scale, while the three item somatic and anxiety scales exhibited lower internal consistency and sensitivity. CONCLUSIONS: These findings provide initial evidence that the BSI-18 is valid for use with homebound older adults. PMID- 20013880 TI - Methionine restriction up-regulates the expression of the pi class of glutathione S-transferase partially via the extracellular signal-regulated kinase-activator protein-1 signaling pathway initiated by glutathione depletion. AB - Understanding the molecular events underlying gene regulation by amino acids has attracted increasing attention. Here, we explored whether the mechanism by which methionine restriction affects the expression of the pi class of glutathione S transferase (GSTP) is related to oxidative stress initiated by glutathione (GSH) depletion. Rat primary hepatocytes were cultured in an L-15-based medium in the absence or presence of 200 muM L-buthionine sulfoximine (BSO) or in a methionine restricted L-15 medium supplemented with 20 muM L-methionine up to 72 h. BSO and methionine restriction time-dependently induced GSTP mRNA and protein expression in a similar pattern accompanied by a decrease in the cellular GSH level. The phosphorylation of extracellular signal-regulated kinase (ERK), but not of c-Jun NH(2)-terminal kinase and p38, was stimulated by methionine restriction and BSO. Electromobility gel shift assay showed that the DNA-binding activity of nuclear activator protein-1 (AP-1) increased in cells exposed to methionine restriction or BSO. With the ERK inhibitor FR180204, AP-1 activation and GSTP expression were abolished. Moreover, the induction of GSTP by methionine restriction and BSO was reversed by GSH monoethyl ester and N-acetylcysteine. Our results suggest that methionine restriction up-regulates GSTP gene expression, which appears to be initiated by the ERK-AP-1 signaling pathway through GSH depletion in rat hepatocytes. PMID- 20013881 TI - Oleuropein and hydroxytyrosol inhibit MCF-7 breast cancer cell proliferation interfering with ERK1/2 activation. AB - The growth of many breast tumors is stimulated by estradiol (E2), which activates a classic mechanism of regulation of gene expression and signal transduction pathways inducing cell proliferation. Polyphenols of natural origin with chemical similarity to estrogen have been shown to interfere with tumor cell proliferation. The aim of this study was to investigate whether hydroxytyrosol (HT) and oleuropein (OL), two polyphenols contained in extra-virgin olive oil, can affect breast cancer cell proliferation interfering with E2-induced molecular mechanisms. Both HT and OL inhibited proliferation of MCF-7 breast cancer cells. Luciferase gene reporter experiments, using a construct containing estrogen responsive elements able to bind estrogen receptor alpha (ERalpha) and the study of the effects of HT or OL on ERalpha expression, demonstrated that HT and OL are not involved in ERalpha-mediated regulation of gene expression. However, further experiments pointed out that both OL and HT determined a clear inhibition of E2 dependent activation of extracellular regulated kinase1/2 belonging to the mitogen activating protein kinase family. Our study demonstrated that HT and OL can have a chemo-preventive role in breast cancer cell proliferation through the inhibition of estrogen-dependent rapid signals involved in uncontrolled tumor cell growth. PMID- 20013882 TI - The metabolic fate of red wine and grape juice polyphenols in humans assessed by metabolomics. AB - The metabolic impact of polyphenol-rich red wine and grape juice consumption in humans was studied using a metabolomics approach. Fifty-eight men and women participated in a placebo-controlled, double-crossover study in which they consumed during a period of 4 wk, either a polyphenol-rich 2:1 dry mix of red wine and red grape juice extracts (MIX) or only a grape juice extract (GJX). Twenty-four-hour urine samples were collected after each intervention. (1)H NMR spectroscopy was applied for global metabolite profiling, while GC-MS was used for focused profiling of urinary phenolic acids. Urine metabolic profiles after intake of both polyphenol-rich extracts were significantly differentiated from placebo using multilevel partial least squares discriminant analysis. A significant 35% increase in hippuric acid excretion (p<0.001) in urine was measured after the MIX consumption as) or only a red grape juice dry extract (GJX). 24-h urine samples were collected after each intervention. 1H-NMR spectroscopy was applied for global metabolite profiling, while gas chromatography-mass spectrometry (GC-MS) was used for focused profiling of urinary phenolic acids. Urine metabolic profiles after intake of both polyphenol rich extracts were significantly differentiated from placebo using multilevel partial least squares discriminant analysis (ML-PLS-DA). A significant 35% increase in hippuric acid excretion (p<0.001) in urine was measured after the MIX consumption compared with placebo, whereas no change was found after GJX consumption. GC-MS-based metabolomics of urine allowed identification of 18 different phenolic acids, which were significantly elevated following intake of either extract. Syringic acid, 3- and 4-hydroxyhippuric acid and 4 hydroxymandelic acid were the strongest urinary markers for both extracts. MIX and GJX consumption had a slightly different effect on the excreted phenolic acid profile and on endogenous metabolite excretion, possibly reflecting their different polyphenol composition. PMID- 20013883 TI - Binding affinity of tea catechins for HSA: characterization by high-performance affinity chromatography with immobilized albumin column. AB - Catechins are the major polyphenols in green tea leaves. Recent studies have suggested that the catechins form complexes with HSA for transport in human blood, and their binding affinity for albumin is believed to modulate their bioavailability. In this study, the binding affinities of catechins and their analogs were evaluated and the relationship between the chemical structure of each catechin and its binding property were investigated. Comparing these catechins by HPLC analysis with the HSA column, we showed that galloylated catechins have higher binding affinities with HSA than non-galloylated catechins. In addition, pyrogallol-type catechins have a high affinity compared to catechol type catechins. Furthermore, the binding affinity of the catechin with 2,3-trans structure was higher than those of the catechin with 2,3-cis structure. The importance of the hydroxyl group on the galloyl group and B-ring was confirmed using methylated catechins. These results indicate that the most important structural element contributing to HSA binding of tea catechins is the galloyl group, followed by the number of hydroxyl groups on the B-ring and the galloyl group or the configuration at C-2. Our findings provide fundamental information on the relationship between the chemical structure of tea catechins and its biological activity. PMID- 20013884 TI - Pyrrolizidine alkaloids in pollen and pollen products. AB - Recently, 1,2-dehydropyrrolizidine alkaloid (PA) ester alkaloids, found predominantly as their N-oxides (PANOs, pyrrolizidine N-oxides), have been reported in both honey and in pollen obtained directly from PA plants and pollen loads collected by bees, raising the possibility of health risks for consumers of these products. We confirm these findings in regard to floral pollen, using pollen collected directly from flowers of the known PA plants Senecio jacobaea, S. vernalis, Echium vulgare and pollinia of Phalaenopsis hybrids, and we extend analyses of 1,2-unsaturated PAs and 1,2-unsaturated PANOs to include bee-pollen products currently being sold in supermarkets and on the Internet as food supplements. PA content of floral pollen ranged from 0.5 to 5 mg/g. The highest values were observed in pollen obtained from Senecio species. Up to 95% of the PAs are found as PANOs. Detailed studies with S. vernalis revealed unique PA patterns in pollen and flowers. While seneciphylline was the most prominent PA in S. vernalis pollen, the flowers were dominated by senecionine. To analyze trace amounts of 1,2-unsaturated PAs in pollen products, our previously elaborated method consisting of strong cation exchange-SPE, two reduction steps followed by silylation and subsequent capillary high-resolution GC-MS using SIM mode was applied. In total, 55 commercially available pollen products were analyzed. Seventeen (31%) samples contained 1,2-unsaturated PAs in the range from 1.08 to 16.35 microg/g, calculated as retronecine equivalents. The 1,2-unsaturated PA content of pollen products is expressed in terms of a single sum parameter and no background information such as foraged plants, pollen analysis, etc. was needed to analyze the samples. The detection limit of overall procedure and the reliable quantitation limit were 0.003 and 0.01 microg/g, respectively. PMID- 20013885 TI - Lupine, a source of new as well as hidden food allergens. AB - The present review summarizes current knowledge about lupine allergy, potential sensitization routes, cross-reactions between lupine and other legumes, and the respective IgE-binding proteins. Since the 1990 s, lupine flour is used as a substitute for or additive to other flours, mostly wheat flour, in several countries of the EU. In 1994, the first case of an immediate-type allergy after ingestion of lupine flour-containing pasta was reported. Since then, the number of published incidents following ingestion or inhalation of lupine flour is rising. So far, the Lupinus angustifolius beta-conglutin has been designated as the allergen Lup an 1 by the International Union of Immunological Societies Allergen Nomenclature Subcommittee. Initially, publications focussed on the fact that peanut-allergic patients were at risk to develop anaphylaxis to lupine due to cross-reactivity between peanut and lupine. At present, however, the ratio between cases of pre-existing legume allergy (mostly peanut allergy) to de novo sensitization to lupine seed is nearly 1:1. Although in December 2006, lupine and products thereof were included in the EU foodstuff allergen list according to the Commission Directive 2006/142/EC amending Annex IIIA of Directive 2000/13/EC in order to prevent severe reactions caused by "hidden food allergens", the majority of patients and medical personnel are still not aware of raw lupine seed as potentially dangerous food allergen. PMID- 20013886 TI - Effect of pistachio oil on gene expression of IFN-induced protein with tetratricopeptide repeats 2: a biomarker of inflammatory response. AB - When incorporated into the diet, pistachios have a beneficial effect on lipid and lipoprotein profiles. However, little is known about potential anti-inflammatory properties. This study was conducted to determine whether pistachio oil and an organic extract from pistachio oil extract (PE) regulated expression of inflammation-related genes. A mouse macrophage cell line (RAW 264.7 cells) was treated with pistachio oil and gene expression microarray analyses were performed. Pistachio oil significantly affected genes involved in immune response, defense response to bacteria, and gene silencing, of which INF-induced protein with tetratricopeptide repeats 2 (Ifit-2) was the most dramatically reduced. PE reduced the LPS-induced Ifit-2 by 78% and the bioactive molecules contained in PE, linoleic acid, and beta-sitosterol recapitulated this inhibition. Promoter analysis identified two adjacent IFN-stimulated response elements, which lie between -110 and -85bp of the 5'-flanking region of the Ifit 2 promoter, as being responsive to LPS activation and inhibition by PE. Our results indicate that pistachio oil and bioactive molecules present therein decrease Ifit-2 expressions, and due to the sensitivity of this effect, this gene is a potential biomarker for monitoring diet-induced changes in inflammation. PMID- 20013887 TI - Resveratrol bioavailability and toxicity in humans. AB - Numerous data are now available on the beneficial properties of the polyphenolic compound resveratrol including its anti-inflammatory and antitumor effects. However, few studies have been performed with resveratrol in humans, and the results of these studies appear fragmentary and sometimes contradictory due to variations in conditions of administration, protocols and methods of assessment. This review article presents the results of recent studies investigating the pharmacokinetics, bioavailability, and toxicity of resveratrol in humans. Resveratrol is well absorbed, rapidly metabolized, mainly into sulfo and glucuronides conjugates which are eliminated in urine. Resveratrol seems to be well tolerated and no marked toxicity was reported. These data are important in the context of human efficacy studies, and they provide further support for the use of resveratrol as a pharmacological drug in human medicine. PMID- 20013888 TI - Effect of dietary fat on endocannabinoids and related mediators: consequences on energy homeostasis, inflammation and mood. AB - Among the several known fatty acid-derived chemical signals, the endogenous ligands of cannabinoid receptors type-1 and -2, two G-protein-coupled receptors involved in several aspects of mammalian physiology and pathology, are perhaps those the levels of which have proven to be most sensitive to the fatty acid composition of the diet. The two most studied such ligands, known as endocannabinoids, are N-arachidonoyl-ethanolamine and 2-archidonoylglycerol, and are found in tissues together with other N-acyl-ethanolamines and 2 acylglycerols, not all of which activate the cannabinoid receptors, although several of them do exhibit important pharmacological effects. In this review article, we describe literature data indicating that the tissue concentrations of the endocannabinoids and related signalling molecules, and hence the activity of the respective receptors, can be modulated by modifying the fatty acid composition of the diet, and particularly its content in long chain PUFAs or in long chain PUFA precursors. We also discuss the potential impact of these diet induced changes of endocannabinoid tone on three of the major pathological conditions in which cannabinoid receptors have been involved, that is metabolic dysfunctions, inflammation and affective disorders. PMID- 20013889 TI - Pyrrolizidine alkaloids (PAs) in honey and pollen-legal regulation of PA levels in food and animal feed required. AB - Pyrrolizidine alkaloids (PAs) are secondary plant constituents that comprise about 400 different structures and occur in two major forms, a tertiary form and the corresponding N-oxide. PAs containing a 1,2-double bond are pre-toxins and metabolically activated by the action of hepatic P-450 enzymes to toxic pyrroles. Besides the acute toxic effects, the genotoxic and tumorigenicity potential of PAs was demonstrated in some eukaryotic model systems. Recently, the potential PA contamination of food and feeding stuff attracted recurrent great deals of attention. Humans are exposed to these toxins by consumption of herbal medicine, herbal teas, dietary supplements or food containing PA plant material. In numerous studies the potential threat to human health by PAs is stated. In pharmaceuticals, the use of these plants is regulated. Considering the PA concentrations observed especially in authentic honey from PA producing plants and pollen products, the results provoke an international regulation of PAs in food. PMID- 20013893 TI - Contrast-enhanced sonography in the diagnosis of acute mesenteric ischemia: case report. AB - Acute mesenteric ischemia is a serious condition associated with high mortality. Multislice CT and magnetic resonance angiography have proved accurate in diagnosing this pathology, which requires a prompt diagnosis to start appropriate therapy. We report the case of an 87-year-old woman with acute mesenteric ischemia, in whom contrast-enhanced sonography visualized the occluded superior mesenteric artery and the infarcted portion of the intestine. PMID- 20013895 TI - microRNAs in acute myeloid leukemia: expression patterns, correlations with genetic and clinical parameters, and prognostic significance. AB - Acute myeloid leukemia (AML) is a malignant disease of hematopoietic cells whose emergence, course, and prognosis is affected by specific recurrent genetic alterations like chromosome aberrations and point mutations, as well as by changes in the expression of certain genes. In the past 2 years, microRNAs (miRNAs)--a novel class of small RNA molecules involved in posttranscriptional gene regulation--have also been shown to be aberrantly expressed in AML. Furthermore, specific miRNA expression patterns were found to be associated with certain genetic and cytogenetic alterations in this disease, and two studies identified miRNAs whose expression levels were predictive of survival. Interestingly, the results of these analyses showed only very limited congruence. This review summarizes published reports on the expression patterns of miRNAs in AML, and discusses possible reasons for the differences in their results. PMID- 20013896 TI - The transcription factor SP1 regulates centriole function and chromosomal stability through a functional interaction with the mammalian target of rapamycin/raptor complex. AB - Specificity protein 1 (SP1) is an essential transcription factor implicated in the regulation of genes that control multiple cellular processes, including cell cycle, apoptosis, and DNA damage. Very few nontranscriptional roles for SP1 have been reported thus far. Using confocal microscopy and centrosome fractionation, we identified SP1 as a centrosomal protein. Sp1-deficient mouse embryonic fibroblasts and cells depleted of SP1 by RNAi have increased centrosome number associated with centriole splitting, decreased microtubule nucleation, chromosome misalignment, formation of multipolar mitotic spindles and micronuclei, and increased incidence of aneuploidy. Using mass spectrometry, we identified P70S6K, an effector of the mTOR/raptor (mTORC1) kinase complex, as a novel interacting protein of SP1. We found that SP1-deficient cells have increased phosphorylation of the P70S6K effector ribosomal protein S6, suggesting that SP1 participates in the regulation of the mTORC1/P70S6K/S6 signaling pathway. We previously reported that aberrant mTORC1 activation leads to supernumerary centrosomes, a phenotype rescued by the mTORC1 inhibitor rapamycin. Similarly, treatment with rapamycin rescued the multiple centrosome phenotype of SP1-deficient cells. Taken together, these data strongly support the hypothesis that SP1 is involved in the control of centrosome number via regulation of the mTORC1 pathway, and predict that loss of SP1 function can lead to aberrant centriole splitting, deregulated mTORC1 signaling, and aneuploidy, thereby contributing to malignant transformation. PMID- 20013897 TI - Focal 9p instability in hematologic neoplasias revealed by comparative genomic hybridization and single-nucleotide polymorphism microarray analyses. AB - Copy number losses in chromosome arm 9p are well-known aberrations in malignancies, including leukemias. The CDKN2A gene is suggested to play a key role in these aberrations. In this study overviewing 9p losses in hematologic neoplasias, we introduce the term focal 9p instability to indicate multiple areas of copy number loss or homozygous loss within a larger heterozygous one in 9p. We have used microarray comparative genomic hybridization to study patients with acute lymphoblastic leukemia (ALL, n = 140), acute myeloid leukemia (n = 50), chronic lymphocytic leukemia (n = 20), and myelodysplastic syndromes (n = 37). Our results show that 9p instability is restricted to ALL. In total, 58/140 (41%) patients with ALL had a loss in 9p. The 9p instability was detected in 19% of the patients with ALL and always included homozygous loss of CDKN2A along with loss of CDKN2B. Other possibly important genes included MTAP, IFN, MLLT3, JAK2, PTPLAD2, and PAX5. 13/27 (48%) patients with the instability had the BCR/ABL1 fusion gene or other oncogene-activating translocation or structural aberrations. Two patients had homozygous loss of hsa-mir -31, a microRNA known to regulate IKZF1. IKZF1 deletion at 7p12.1 was seen in 10 (37%) patients with the 9p instability. These findings suggest that, in ALL leukemogenesis, loss of CDKN2A and other target genes in the instability region is frequently associated with BCR/ABL1 and IKZF1 dysfunction. The multiple mechanisms leading to 9p instability including physical or epigenetic loss of the target genes, loss of the microRNA cluster, and the role of FRA9G fragile site are discussed. PMID- 20013898 TI - Application of nonparametric quantile regression to body mass index percentile curves from survey data. AB - Increasing rates of overweight among children in the U.S. stimulated interest in obtaining national percentile curves of body size to serve as a benchmark in assessing growth development in clinical and population settings. In 2000, the U.S. Centers for Disease Control and Prevention (CDC) developed conditional percentile curves for body mass index (BMI) for ages 2-20 years. The 2000 CDC BMI for-age curves are partially parametric and only partially incorporated the survey sample weights in the curve estimation. As a result, they may not fully reflect the underlying pattern of BMI-for-age in the population. This motivated us to develop a nonparametric double-kernel-based method and automatic bandwidth selection procedure. We include sample weights in the bandwidth selection, conduct median correction to reduce small-sample smoothing bias, and rescale the bandwidth to make it scale invariant. Using this procedure we re-estimate the national percentile BMI-for-age curves and the prevalence of high-BMI children in the U.S. PMID- 20013902 TI - DNA manipulation by means of insulator-based dielectrophoresis employing direct current electric fields. AB - Electrokinetic techniques offer a great potential for biological particle manipulation. Among these, dielectrophoresis (DEP) has been successfully utilized for the concentration of bioparticles. Traditionally, DEP is performed employing microelectrodes, an approach with attractive characteristics but expensive due to microelectrode fabrication costs. An alternative is insulator-based DEP, a method where non-uniform electric fields are created with arrays of insulating structures. This study presents the concentration of linear DNA particles (pET28b) employing a microchannel, with an array of cylindrical insulating structures and direct current electric fields. Results showed manipulation of DNA particles with a combination of electroosmotic, electrophoretic, and dielectrophoretic forces. Employing suspending media with conductivity of 104 muS/cm and pH of 11.15, under applied fields between 500 and 1500 V/cm, DNA particles were observed to be immobilized due to negative dielectrophoretic trapping. The observation of DNA aggregates that occurred at higher applied fields, and dispersed once the field was removed is also included. Finally, concentration factors varying from 8 to 24 times the feed concentration were measured at 2000 V/cm after concentration time-periods of 20-40 s. The results presented here demonstrate the potential of insulator-based DEP for DNA concentration, and open the possibility for fast DNA manipulation for laboratory and large-scale applications. PMID- 20013903 TI - Electrophoresis microchip with integrated waveguides for simultaneous native UV fluorescence and absorbance detection. AB - Simultaneous label-free detection of UV absorbance and native UV-excited fluorescence in an electrophoresis microchip is presented. UV transparent integrated waveguides launch light at a wavelength of 254 nm from a mercury lamp along the length of a 1-mm long detection cell. Transmitted UV light is collected by another waveguide in the opposite end of the detection cell, while visible fluorescence is collected vertically through the lid of the chip. The background of scattered excitation light is suppressed by detection perpendicular to the excitation, the limited UV transparency of the borosilicate lid and by choosing a PMT insensitive to the excitation light. This way, the need for a fluorescence filter is eliminated. Calibration curves were measured for serotonin, tryptophan, propranolol and acetaminophen, and separations of the four compounds were demonstrated by electrophoresis and MEKC. All compounds could be detected in the micromolar range by absorbance detection, but fluorescence detection improved detection limits for compounds displaying native UV fluorescence up to ten times. The simultaneous detection also proved useful for the identification of compounds with similar retention times and even enables accurate quantification of co eluting compounds. PMID- 20013904 TI - Temperature gradient focusing in miniaturized free-flow electrophoresis devices. AB - Temperature gradient focusing is a method to separate and focus any charged analytes even without accessible isoelectric point, and has been already widely used in CE. In this paper, we demonstrate the application of temperature gradient focusing to free-flow electrophoresis. Besides focusing and separation experiments of proteins, the stability of the temperature gradient under flow conditions and the temperature dependence of fluorescence dyes have also been investigated. PMID- 20013905 TI - Pump-free and low-cost negative pressure sampling device for rapid sample loading in MCE. AB - A pump-free and low-cost negative pressure sampling device for injecting well defined non-biased sample plugs into the separation channel of MCE was developed. It was composed of a pipet bulb, a 3-way electromagnetic valve and a single voltage supply at constant voltage. A sub-atmospheric pressure was created by hand-pressing air out of the pipet bulb and retained in it by switching the 3-way electromagnetic valve at cutoff position. During the sample loading stage, the sub-atmospheric pressure in the pipet bulb was applied via a 3-way electromagnetic valve to the headspace of the sealed sample waste reservoir (SW). A pinched sample plug was formed at the channel intersection in less than 0.5 s. Once the 3-way electromagnetic valve was switched to connect SW to ambient atmosphere to release the vacuum in SW, electrophoresis separation was consequently activated under the electric potentials applied. Experimental results demonstrated the pump-free negative pressure sampling device worked well in a wide vacuum degree ranged from -250 to -30 mbar with a satisfactory analytical precision. The sample consumption for each cycle was calculated to be 51-12 nL under the sampling pressure. Theoretical deduction indicates that the volume of the pipet bulb can be further reduced to 1 mL, which is critical for minimizing the sampling device for MCE. PMID- 20013906 TI - Dry film microchips for miniaturised separations. AB - In this work microfluidic devices were made from the dry film photoresist Ordyl SY330, characterised by optical and electron microscopy and used for electrophoretic separations. A simple and fast microfabrication process was developed for the fabrication of channels that are 50 microm wide and 30 microm in height, requiring only the use of an office laminator, a hot plate, an exposure source and mask and an electric drill to make four microdevices in less than 1 h. The optical properties of the photoresist were studied and the resist showed significant absorbance below 370 nm and 570-630 nm, and had an optical transmission of 80% between 400 and 550 nm. Fluorescence emission over the region of maximum transmission was low allowing these devices to be used for fluorescence detection at 488/512 nm. Electrophoretic separation of APTS and three derivatised sugars was performed in 20 mM phosphate buffer, pH 2.5 with efficiencies of the three sugars of 40,000 plates (2,100,000 plates/m) within 30 s at a field strength of 500 V/cm. The simple fabrication process also allowed microchannels to be easily filled with chromatography particles before sealing, avoiding the challenging task of slurry packing, and the potential of these devices for liquid chromatography was demonstrated by the extraction of fluorescein onto anion exchange particles. PMID- 20013907 TI - Fabrication of PMMA microfluidic chips using disposable agar hydrogel templates. AB - A novel method based on disposable agar hydrogel temples have been developed for the fabrication of PMMA microfluidic chips. Molten agar hydrogel was sandwiched between a glass plate and a PMMA template bearing negative relief of microstructure. After cooling, the negative PMMA template could be easily separated from the solidified agar hydrogel and a layer of agar hydrogel temple bearing high-fidelity positive relief of the microstructure was left on the glass plate. Prepolymerized methyl methacrylate molding solution containing ultraviolet initiator was subsequently sandwiched between the agar hydrogel template and a PMMA plate and was allowed to polymerize under ultraviolet light to fabricate the PMMA channel plate at room temperature. Complete microchips could be obtained by bonding the channel plates with covers. The prepared microfluidic microchips have been successfully employed in the electrophoresis separation and detection of several ions in connection with contactless conductivity detection. PMID- 20013908 TI - Graft linker immobilization for spatial control of protein immobilization inside fused microchips. AB - Fused silica glass microchips have several attractive features for lab-on-a-chip applications; they can be machined with excellent precision down to nanospace; are stable; transparent and can be modified with a range of silanization agents to change channel surface properties. For immobilization, however, ligands must be added after bonding, since the harsh bonding conditions using heat or hydrofluoric acid would remove all prior immobilized ligands. For spatial control over immobilization, UV-mediated immobilization offers several advantages; spots can be created in parallel, the feature size can be made small, and spatial control over patterns and positions is excellent. However, UV sensitive groups are often based on hydrophobic chemical moieties, which unfortunately result in greater non-specific binding of biomolecules, especially proteins. Here, we present techniques in which any -CH(x) (x=1,2,3) containing surface coating can be used as foundation for grafting a hydrophilic linker with a chemical anchor, a carboxyl group, to which proteins and amine containing molecules can be covalently coupled. Hence, the attractive features of many well-known protein and biomolecule repelling polymer coatings can be utilized while achieving site specific immobilization only to pre-determined areas within the bonded microchips. PMID- 20013909 TI - Evaluation of microchip material and surface treatment options for IEF of allergenic milk proteins on microchips. AB - The use of glass and PDMS microchips has been investigated to perform rapid and efficient separation of allergenic whey proteins by IEF. To decrease EOF and to limit protein adsorption, two coating procedures have been compared. The first one consists in immobilizing hydroxypropyl cellulose (HPC) and the second one poly(dimethylacrylamide-co-allyl glycidyl ether) (PDMA-AGE). EOF limitation has been evaluated using frontal electrophoresis of a fluorescent marker of known effective mobility. EOF velocity was decreased by a factor about 100 and 30, respectively. pH gradient formation has been evaluated for each microchip using fluorescent pI markers. It was demonstrated that as expected a coating was essential to avoid pH gradient drift. Both coatings were efficient on glass microchips, but only PDMA-AGE allowed satisfying focusing of pI markers on PDMS microchips. Fluorescent covalent and noncovalent labelings of milk proteins have been compared by IEF on slab-gels. IEF separation of three major allergenic whey proteins [beta-lactoglobulin A (pI 5.25) and B (pI 5.35) and alpha-lactalbumin (pI 4.2-4.5)] was performed in both microchips. Milk proteins were separated with better resolution and shorter analysis time than by classical CIEF. Finally, better resolutions for milk allergens separation were obtained on glass microchips. PMID- 20013910 TI - Microchip CGE linked to immunoprecipitation as an alternative to Western blotting. AB - A novel approach for protein identification is presented, which combines the specificity of an immunoprecipitation approach with the sensitivity of protein detection in microchip CGE. This method involves derivatization of the sample proteins with a fluorescent dye, target protein isolation with specific antibodies and Protein A coated magnetic beads, and automated sizing and quantification of the eluted samples on microchips. The performance of the new technique was demonstrated with glutathion-S-transferase- and polyHistidine tagged target proteins in an Escherichia coli background. A specific detection of target proteins was possible down to 0.001% or 1 ng target protein in a background of 100 microg E. coli protein. With this approach, proteins ranging from 10 to 220 kDa could be identified with a panel of different target-specific antibodies. The reproducibility of the method was very similar to standard microchip CGE methods. In a direct comparison to Western blotting, a similar sensitivity and specificity of both techniques was observed. However, the new approach compares favorably to Western blotting in terms of time-to-result, usability and labor intensity, antibody consumption and access to quantitative data. PMID- 20013911 TI - Rapid discrimination of single-nucleotide mismatches based on reciprocating flow on a compact disc microfluidic device. AB - In this study, a compact disc (CD) microfluidic device based on reciprocating flow is developed for the rapid discrimination of single-nucleotide mismatch (SNM). This device consists of a PDMS CD slab containing eight SNM assay units and a glass substrate immobilized with DNA probe array. The rapid SNM assay is realized by two-step controllable reciprocating flow in the microchannel, resulting from centrifugal force and capillary action as well. A perfect-match probe and three mismatched probes containing a SNM at various positions were designed to investigate the ability of this device to discriminate SNM. Based on the difference between the dissociation kinetics of perfect-match and mismatched duplexes, this CD device was able to discriminate SNM within 400 s. Moreover, only sub-microliter volume DNA samples (350 nL) and washing buffers (350 nL) were required per assay unit. The results indicate that this device has the potential to be used for fast, accurate gene diagnostics, especially for the detection of SNPs. PMID- 20013912 TI - Comparing polyelectrolyte multilayer-coated PMMA microfluidic devices and glass microchips for electrophoretic separations. AB - There is a continuing drive in microfluidics to transfer microchip systems from the more expensive glass microchips to cheaper polymer microchips. Here, we investigate using polyelectrolyte multilayers (PEM) as a coating system for PMMA microchips to improve their functionality. The multilayer system was prepared by layer-to-layer deposition of poly(diallyldimethylammonium) chloride and polystyrene sulfonate. Practical aspects of coating PMMA microchips were explored. The multilayer buildup process was monitored using EOF measurements, and the stability of the PEM was investigated. The performance of the PEM-PMMA microchip was compared with those of a standard glass microchip and a PEM-glass microchip in terms of EOF and separating two fluorescent dyes. Several key findings in the development of the multilayer coating procedure for PMMA chips are also presented. It was found that, with careful preparation, a PEM-PMMA microchip can be prepared that has properties comparable--and in some cases superior--to those of a standard glass microchip. PMID- 20013913 TI - High-fidelity fractionation of ssDNA fragments differing in size by one-base on a spiral-channel electrophoretic chip. AB - For the fractionation of fragments of interest from selective PCR products generated by high coverage gene expression profiling (HiCEP) analysis, high resolution with the ability to discriminate and fractionate fragments differing by one base (base pair) in size is highly required. We report here on a new 4 inch diameter spiral-channel chip device for automatic high-fidelity fractionation. Overlapping DNA fragments of 180, 181 and 182 bases, with only one base difference in size, were successfully fractionated. The collected fragments were PCR amplified, and then evaluated by size checking analysis, DNA sequencing, and homolog search. The high-resolution fractionation has been achieved because of the combined contributions of (i) the high-resolution separation using a 30 cm long spiral channel, (ii) a blocking technique to avoid contamination from unselected fragments during CE, and (iii) precise micro-scale target extraction. Contaminations due to unselected fractions have been greatly decreased to a negligible level by optimization of the extraction position and extraction time corresponding to the targeted segment only. This technique can be adapted to a wide range of applications, such as protein or cell collections where requirements for the high purity are more important than the amount of recovered fractionated material. PMID- 20013914 TI - A microfluidic device for characterizing the invasion of cancer cells in 3-D matrix. AB - A microfluidic device was developed for the study of directed invasion of cancer cells in 3-D matrix with concentration gradient. This device consists of two parallel perfusion channels connected by two cell culture chambers. To mimic extracellular matrix (ECM), gelled basement membrane extract (BME) was used to support 3-D distribution of breast cancer cells (MCF7) in cell culture chambers. A stable linear concentration gradient of epidermal growth factor (EGF) was generated across the chambers by continuous perfusion. Using the device, we investigated MCF7 cell invasion induced by different concentrations of EGF in 3-D matrix. It was found that cancer cells responded to EGF stimulation with forming cellular protrusions and migrating towards high EGF concentration. We further investigated the anti-invasion effect of GM 6001, a matrix metalloproteinase inhibitor. We identified that matrix metalloproteinase inhibition repressed both cellular protrusion formation and cell migration in 3-D matrix. These findings suggest that EGF is able to induce MCF7 cell invasion in 3-D extracellular matrix and this effect is dependent on proteolytic activity. This device is relatively simple to construct and operate. It should be a useful platform for elucidating the mechanism of cancer invasion and screening anti-invasion drugs for cancer therapy. PMID- 20013915 TI - Chip electrophoretic characterization of liposomes with biological lipid composition: Coming closer to a model for viral infection. AB - In first attempts at elucidating the transfer of the RNA genome of a human Rhinovirus through lipid membranes in vitro we made use of liposomes decorated with recombinant receptors. This model system was characterized previously by CE but suffered from the requirement for inclusion of polyethylene glycol-modified lipids for reliable separations [Weiss, V. U., Bilek, G., Pickl-Herk, A., Blaas, D., Kenndler, E., Electrophoresis 2009, 30, 2123-2128.]. We here report the analysis of liposomes with a lipid composition much more similar to that of biological lipid bilayers. We found that vesicles containing and lacking this non physiologic lipid differ significantly in their electrophoretic mobility (by factor 2) although the concentration of charge-bearing lipids in their bilayers is the same. We demonstrate that binding of a human Rhinovirus to the latter liposomes decorated with a cognate receptor can be analysed via electrophoresis on microchips; we support our results with transmission electron microscopy. PMID- 20013917 TI - Feasibility of randomized controlled trials in liver surgery using surgery related mortality or morbidity as endpoint (Br J Surg 2009; 96: 1005-1014). PMID- 20013919 TI - Factors influencing survival after complete resection of pulmonary metastases from colorectal cancer (Br J Surg 2009; 96: 1058-1065). PMID- 20013921 TI - Blumgart anastomosis for pancreaticojejunostomy minimizes severe complications after pancreatic head resection (Br J Surg 2009; 96: 741-750). PMID- 20013924 TI - Commentary. PMID- 20013925 TI - Commentary. PMID- 20013926 TI - Meta-analysis of randomized controlled trials comparing laparoscopic with open mesh repair of recurrent inguinal hernia. AB - BACKGROUND: Although there is plentiful evidence regarding the use of laparoscopic surgery for primary inguinal hernia, there is a paucity of literature concerning its role after recurrence. There has been no quantitative review of the evidence, despite suggestions that pooled analysis of existing data is required. METHODS: Medline, Embase, trial registries, conference proceedings and reference lists were searched for controlled trials of laparoscopic versus conventional open surgery for mesh repair of recurrent hernia. The primary outcomes were recurrence and chronic pain. Secondary outcomes were operating time, visual analogue pain score, superficial wound infection, haematoma or seroma formation, time to return to normal activities and serious complications requiring operation. Pooled odds ratios were calculated for categorical outcomes and weighted mean differences for continuous outcomes. RESULTS: Four trials were included in the analysis. There was no effect on recurrence or chronic pain. Laparoscopic surgery was associated with significantly less postoperative pain, a quicker return to normal activities and fewer wound infections, at the cost of a longer operating time. There was no difference in haematoma formation or the need for additional operations. CONCLUSION: Careful patient selection and surgeons' experience are important in the selection of technique for recurrent inguinal hernia repair. PMID- 20013927 TI - Randomized clinical trial of co-amoxiclav versus no antibiotic prophylaxis in varicose vein surgery. AB - BACKGROUND: Wound infection rates of up to 16 per cent are reported following varicose vein surgery and the value of antibiotic prophylaxis in clean surgery remains unclear. METHODS: Some 443 patients undergoing varicose vein surgery randomly received a single prophylactic dose of 1.2 g co-amoxiclav (219 patients) or no antibiotic (224). Patients completed a wound diary on postoperative days 3, 5, 7, 9 and 10 using an adapted ASEPSIS method of wound assessment, and were reviewed after 14 days. RESULTS: Patients who had prophylaxis had lower ASEPSIS wound scores on days 3, 5 and 7 (P = 0.043, P = 0.032 and P = 0.003 respectively), and lower total ASEPSIS scores (median (interquartile range) 3 (0 9) versus 6 (0-15); P = 0.013). They were less likely to consult their general practitioner (16.0 versus 24.3 per cent; P = 0.040) or to receive postoperative antibiotics (4.7 versus 13.5 per cent; P = 0.002) for wound-related problems. Wound outcomes were worse with higher body mass index (odds ratio (OR) 0.92 (95 per cent confidence interval (c.i.) 0.87 to 0.97); P = 0.005) and current smoking (OR 0.5 (0.3 to 0.9); P = 0.033). Prophylactic antibiotics conferred satisfactory wound healing (OR 2.2 (95 per cent c.i. 1.3 to 3.6); P = 0.003). CONCLUSION: Antibiotic prophylaxis reduced wound-related problems after varicose vein surgery. Registration number ISRCTN12467340 (http://www.controlled-trials.com). PMID- 20013928 TI - Liver resection under total vascular exclusion with or without preceding Pringle manoeuvre. AB - BACKGROUND: Adequate control of bleeding is crucial during liver resection. This study analysed the safety and efficacy of hepatectomy under total hepatic vascular exclusion (THVE) in patients with tumours encroaching or infiltrating the hepatic veins and/or the inferior vena cava (IVC). METHODS: All patients undergoing liver resection with THVE between January 2000 and July 2006 were identified from a prospectively collected database containing 2400 patients. Data on patient demographics, surgical procedure and outcome were collected. RESULTS: A total of 87 patients scheduled for liver resection under THVE were identified, 77 with malignant tumours and ten with benign disease. THVE could not be used in two patients (2 per cent) owing to haemodynamic intolerance during trial clamping. Seventeen patients received simultaneous clamping of the portal triad and vena cava, and 68 had portal triad clamping followed by concomitant portal and vena cava clamping. The mean(s.d.) duration of THVE was 28.3(7.5) and 18.7(5.2) min respectively. Overall postoperative complication and operative mortality rates were 53 and 2 per cent respectively. Mean(s.d.) hospital stay was 16.8(4.7) days. CONCLUSION: Major hepatic resection for tumours encroaching on the hepatic veins or IVC can be carried out under THVE with reasonable morbidity and mortality. PMID- 20013929 TI - Diagnosis and treatment of spontaneous group A streptococcal peritonitis. AB - BACKGROUND: Primary group A streptococcal peritonitis (PSAP) is a rare, fulminant and often fatal infection. The clinical manifestations include diffuse peritoneal signs with toxic shock syndrome and sometimes fasciitis. METHODS: Patients with PSAP diagnosed between December 2002 and December 2006 were studied retrospectively, focusing on the initial presentation, diagnosis, treatment and outcome. RESULTS: Six patients were identified (five women and one man). The clinical presentation was heterogeneous. All six patients had diffuse peritonitis, four had toxic shock syndrome on hospital admission and two patients also had fasciitis. All patients were treated surgically, and the final diagnosis was confirmed after operation. There were no deaths, but two patients had aesthetic sequelae owing to necrotizing fasciitis. CONCLUSION: PSAP is a rare condition, often requiring aggressive surgical treatment. Group A streptococcal peritonitis should be suspected in patients with no radiological evidence of a peritoneal portal of entry and no history of ascites. PMID- 20013930 TI - Clinical outcomes of ileorectal anastomosis for ulcerative colitis. AB - BACKGROUND: The aim of this study was to determine the fate of the rectum, functional results and quality of life after ileorectal anastomosis (IRA) in ulcerative colitis. METHODS: Patients with ulcerative colitis and indeterminate colitis who underwent IRA from 1971 to 2006 were evaluated retrospectively. Twenty-two patients with an IRA were matched by age, sex and follow-up duration with 66 patients with an ileal pouch-anal anastomosis (IPAA) and compared for functional outcomes and quality of life. RESULTS: Eighty-six patients with an IRA were included. Median follow-up was 9 (range 1-36) years. Rectal dysplasia and cancer rates were 17 and 8 per cent respectively. The rectum was resected in 46 patients (53 per cent) because of refractory proctitis in 24, rectal dysplasia in 15 and rectal cancer in seven. The cumulative probability of having a functioning IRA at 10 and 20 years was 74 and 46 per cent respectively. Patients with an IRA had fewer bowel movements (P = 0.020) and less night-time seepage (P = 0.020) but increased urgency (P < 0.001) compared with patients with an IPAA, whereas quality of life was similar. CONCLUSION: In selected patients with ulcerative colitis IRA gives an acceptable quality of life and functional outcome that are comparable to those in patients with an IPAA. Owing to the risk of cancer, surveillance of the rectum is mandatory. PMID- 20013931 TI - Risk modelling of outcome after general and trauma surgery (the IRIS score). AB - BACKGROUND: A practical, easy to use model was developed to stratify risk groups in surgical patients: the Identification of Risk In Surgical patients (IRIS) score. METHODS: Over 15 years an extensive database was constructed in a general surgery unit, containing all patients who underwent general or trauma surgery. A logistic regression model was developed to predict mortality. This model was simplified to the IRIS score to enhance practicality. Receiver operating characteristic (ROC) curve analysis was performed. RESULTS: The database contained a consecutive series of 33 224 patients undergoing surgery. Logistic regression analysis gave the following formula for the probability of mortality: P (mortality) = A/(1 + A), where A = exp (-4.58 + (0.26 x acute admission) + (0.63 x acute operation) + (0.044 x age) + (0.34 x severity of surgery)). The area under the ROC curve (AUC) was 0.92. The IRIS score also included age (divided into quartiles, 0-3 points), acute admission, acute operation and grade of surgery. The AUC predicting postoperative mortality was 0.90. CONCLUSION: The IRIS score accurately predicted mortality after general or trauma surgery. PMID- 20013932 TI - A major trauma centre is a specialty hospital not a hospital of specialties. AB - BACKGROUND: High estimates of preventable death rates have renewed the impetus for national regionalization of trauma care. Institution of a specialist multidisciplinary trauma service and performance improvement programme was hypothesized to have resulted in improved outcomes for severely injured patients. METHODS: This was a comparative analysis of data from the Royal London Hospital (RLH) trauma registry and Trauma Audit and Research Network (England and Wales), 2000-2005. Preventable mortality was evaluated by prospective analysis of the RLH performance improvement programme. RESULTS: Mortality from critical injury at the RLH was 48 per cent lower in 2005 than 2000 (17.9 versus 34.2 per cent; P = 0.001). Overall mortality rates were unchanged for acute hospitals (4.3 versus 4.4 per cent) and other multispecialty hospitals (8.7 versus 7.3 per cent). Secondary transfer mortality in critically injured patients was 53 per cent lower in the regional network than the national average (5.2 versus 11.0 per cent; P = 0.001). Preventable death rates fell from 9 to 2 per cent (P = 0.040) and significant gains were made in critical care and ward bed utilization. CONCLUSION: Institution of a specialist trauma service and performance improvement programme was associated with significant improvements in outcomes that exceeded national variations. PMID- 20013933 TI - Nutritional risk is a clinical predictor of postoperative mortality and morbidity in surgery for colorectal cancer. AB - BACKGROUND: This study investigated whether nutritional risk scores applied at hospital admission predict mortality and complications after colorectal cancer surgery. METHODS: Some 186 patients were studied prospectively. Clinical details, Reilly's Nutrition Risk Score (NRS) and Nutritional Risk Screening 2002 (NRS 2002) score, tumour stage and surgical procedure were recorded. RESULTS: The prevalence of patients at nutritional risk was 31.7 per cent according to Reilly's NRS and 39.3 per cent based on the NRS-2002. Such patients had a higher mortality rate than those not at risk according to Reilly's NRS (8 versus 1.6 per cent; P = 0.033), but not the NRS-2002 (7 versus 1.8 per cent; P = 0.085). Based on the NRS-2002, there was a significant difference in postoperative complication rate between patients at nutritional risk and those not at risk (62 versus 39.8 per cent; P = 0.004) but not if Reilly's NRS was used (58 versus 44.1 per cent; P = 0.086). Nutritional risk was identified as an independent predictor of postoperative complications (odds ratio 2.79; P = 0.002). CONCLUSION: Nutritional risk screening may be able to predict mortality and morbidity after surgery for colorectal cancer. However, the diverse results reflect either the imprecision of the tests or the small sample size. PMID- 20013934 TI - Population-based trend analysis of 2813 patients undergoing laparoscopic sigmoid resection. AB - BACKGROUND: The use of laparoscopic sigmoid resection for diverticular disease has become increasingly popular. The objective of this trend analysis was to assess whether clinical outcomes following laparoscopic sigmoid resection for diverticular disease have improved over the past 10 years. METHODS: The analysis was based on the prospective database of the Swiss Association of Laparoscopic and Thoracoscopic Surgery. Some 2813 patients undergoing elective laparoscopic sigmoid resection for diverticular disease from 1995 to 2006 were included. Unadjusted and risk-adjusted analyses were performed. RESULTS: Over time, there was a significant reduction in the conversion rate (from 27.3 to 8.6 per cent; P(trend) < 0.001), local postoperative complication rate (23.6 to 6.2 per cent; P(trend) = 0.004), general postoperative complication rate (14.6 to 4.9 per cent; P(trend) = 0.024) and reoperation rate (5.5 to 0.6 per cent; P(trend) = 0.015). Postoperative median length of hospital stay significantly decreased from 11 to 7 days (P(trend) < 0.001). CONCLUSION: This first trend analysis in the literature of clinical outcomes after laparoscopic sigmoid resection, based on almost 3000 patients, has provided compelling evidence that rates of postoperative complications, conversion and reoperation, and length of hospital stay have decreased significantly over the past 10 years. PMID- 20013935 TI - Importance of clear resection margins in anorectal malignant melanoma. AB - BACKGROUND: Anorectal melanoma is rare and surgery is the recommended primary treatment. There has been some debate whether abdominoperineal resection (APR) or local excision is most appropriate. The aim of this study was to provide a population-based analysis of symptoms, treatment and outcome. METHODS: From the Swedish National Cancer Registry, 251 patients with anorectal melanoma were identified from 1960 to 1999. Medical reports were collected and reviewed retrospectively. R0 resection was defined by clear macroscopic margins and a pathology report showing a margin greater than 10 mm. Survival was compared with the log rank test, and Cox multivariable analysis was performed. RESULTS: APR and local excision was performed in 66 and 86 patients respectively. Median survival after surgery was 14 months, with no statistically significant difference between the two groups. Seventy-two patients in whom an R0 resection was achieved, irrespective of approach, had a significantly better overall 5-year survival rate than patients with involved margins (19 versus 6 per cent; P < 0.001). Multivariable analysis showed resection status and tumour stage to be independent prognostic variables. CONCLUSION: Both APR and LE seem appropriate for anorectal melanoma provided clear margins can be achieved; prognosis is poor regardless of surgical approach. PMID- 20013936 TI - Adhesions and incisional hernias following laparoscopic versus open surgery for colorectal cancer in the CLASICC trial. AB - BACKGROUND: This study investigated adhesive intestinal obstruction (AIO) and incisional hernia (IH) in patients undergoing laparoscopically assisted and open surgery for colorectal cancer. METHODS: In a case-note review of patients randomized to the Medical Research Council's Conventional versus Laparoscopic Assisted Surgery In Colorectal Cancer (CLASICC) trial, primary and key secondary endpoints were AIO and IH admission rates respectively. RESULTS: Of 411 patients, 11 were admitted for AIO: four (3.1 per cent) of 131 patients in the open arm of the trial versus seven (2.5 per cent) of 280 in the laparoscopic arm (difference 0.6 (95 per cent confidence interval (c.i.) - 2.9 to 4.0) per cent). Thirty-six patients developed IH: 12 (9.2 per cent) after open versus 24 (8.6 per cent) after laparoscopic surgery (difference 0.6 (95 per cent c.i. - 5.3 to 6.5) per cent). Results by actual procedure showed higher AIO and IH rates in the 24.5 per cent of patients who converted from laparoscopic to open surgery (AIO: 2.3, 2.0 and 6 per cent; IH: 8.6, 7.4 and 11 per cent-for open, laparoscopic and converted operations respectively). CONCLUSION: Although this study has not confirmed that laparoscopic surgery reduces rates of AIO and IH after colorectal cancer surgery, trends suggest that a reduction in conversion to open surgery and elimination of port-site hernias may produce such an effect. Registration number for CLASICC trial: ISRCTN74883561 (http://www.controlled-trials.com). PMID- 20013937 TI - Real longitudinal data analysis for real people: building a good enough mixed model. AB - Mixed effects models have become very popular, especially for the analysis of longitudinal data. One challenge is how to build a good enough mixed effects model. In this paper, we suggest a systematic strategy for addressing this challenge and introduce easily implemented practical advice to build mixed effects models. A general discussion of the scientific strategies motivates the recommended five-step procedure for model fitting. The need to model both the mean structure (the fixed effects) and the covariance structure (the random effects and residual error) creates the fundamental flexibility and complexity. Some very practical recommendations help to conquer the complexity. Centering, scaling, and full-rank coding of all the predictor variables radically improve the chances of convergence, computing speed, and numerical accuracy. Applying computational and assumption diagnostics from univariate linear models to mixed model data greatly helps to detect and solve the related computational problems. Applying computational and assumption diagnostics from the univariate linear models to the mixed model data can radically improve the chances of convergence, computing speed, and numerical accuracy. The approach helps to fit more general covariance models, a crucial step in selecting a credible covariance model needed for defensible inference. A detailed demonstration of the recommended strategy is based on data from a published study of a randomized trial of a multicomponent intervention to prevent young adolescents' alcohol use. The discussion highlights a need for additional covariance and inference tools for mixed models. The discussion also highlights the need for improving how scientists and statisticians teach and review the process of finding a good enough mixed model. PMID- 20013938 TI - Sleep-wake functioning along the cancer continuum: focus group results from the Patient-Reported Outcomes Measurement Information System (PROMIS((r))). AB - OBJECTIVE: Cancer and its treatments disturb sleep-wake functioning; however, there is little information available on the characteristics and consequences of sleep problems associated with cancer. As part of an effort to improve measurement of sleep-wake functioning, we explored the scope of difficulties with sleep in a diverse group of patients diagnosed with cancer. METHODS: We conducted 10 focus groups with patients recruited from the Duke University tumor registry and oncology/hematology clinics. Separate groups were held with patients scheduled to begin or currently undergoing treatment for breast, prostate, lung, colorectal, hematological, and other cancer types and with patients who were in posttreatment follow-up. The content of the focus group discussions was transcribed and analyzed for major themes by independent coders. RESULTS: Participants not only reported causes of sleep disturbance common in other populations, such as pain and restless legs, but they also reported causes that may be unique to cancer populations, including abnormal dreams, anxiety about cancer diagnosis and recurrence, night sweats, and problems with sleep positioning. Many participants felt that sleep problems reduced their productivity, concentration, social interactions, and overall quality of life. Many also shared beliefs about the increased importance of sleep when fighting cancer. CONCLUSIONS: The findings underscore the need for interventions that minimize the negative impact of cancer and its treatments on sleep. This study will inform efforts now underway to develop a patient-reported measure of sleep wake functioning that reflects the breadth of concepts considered important by patients with cancer. PMID- 20013939 TI - miR133a regulates cardiomyocyte hypertrophy in diabetes. AB - BACKGROUND: Diabetic cardiomyopathy, characterized by cardiac hypertrophy and contractile dysfunction, eventually leads to heart failure. We have previously shown that alterations of a number of key molecules are involved in producing cardiomyocyte hypertrophy in diabetes. The aim of the present study was to determine whether microRNAs (miRNA) play a role in mediating altered gene expression and structural/functional deficits in the heart in diabetes. METHODS: STZ-induced diabetic mice were haemodynamically investigated after 2 months of diabetes to establish the development of cardiomyopathy. The tissues were then examined for gene expression and microRNA analysis. We further investigated neonatal rat cardiomyocytes to identify the mechanisms of glucose-induced hypertrophy and the potential role of miR133a. RESULTS: Diabetic mice showed myocardial contractile dysfunction and augmented mRNA expression of atrial and brain natriuretic peptides (ANP, BNP), MEF2A and MEF2C, SGK1 and IGF1R compared to age- and sex-matched controls. Cardiac tissues from these mice showed alteration of multiple miRNAs by array analysis including miR133a, which was confirmed by RT-PCR. In vitro exposure of cardiomyocytes to high levels of glucose produced hypertrophic changes and reduced expression of miRNA133a. Finally, transfection of miR133a mimics prevented altered gene expression and hypertrophic changes. CONCLUSION: Data from these studies demonstrate a novel glucose-induced mechanism regulating gene expression and cardiomyocyte hypertrophy in diabetes which is mediated through miR133a. PMID- 20013940 TI - Growth rates of small abdominal aortic aneurysms correlate with clinical events. AB - BACKGROUND: This retrospective analysis of prospectively collected abdominal aortic aneurysm (AAA) screening data aimed to identify predictors of AAA-related events (surgery or death) with a view to better targeting of screening. METHODS: For the interval 1984-2007, data for 1649 subjects with an AAA were collected prospectively as part of the Chichester AAA screening programme. This included serial aortic size measurements, blood pressure, risk factors for arterial disease and concurrent medications. AAA growth rates were adjusted for risk factor confounders using flexible hierarchical modelling. AAA growth distribution was analysed using Silverman's test of multimodality. RESULTS: Some 1231 subjects met the inclusion criteria of having more than one scan and a surveillance interval of over 3 months. AAA growth showed a bimodal pattern with nearly 50 per cent of all aneurysms never progressing to surgery or rupture. Adjusted annual AAA growth rates of at least 2 mm significantly predicted AAA-related events. CONCLUSION: This analysis identified a bimodal growth pattern for AAA, with a significant association between annual AAA growth rate of at least 2 mm and AAA related events. PMID- 20013941 TI - Aspects of observing and claiming allele flips in association studies. AB - Significant allele flipping, where associations for the same disease occur at opposite alleles of the same bi-allelic locus, is increasing. But when is a significant allele flip genuine? We address the statistical issues of claiming and observing genuine allele flips in actual samples. We show that unless an allele flip is genuine, the probability of observing a significant allele flip in samples ascertained similarly from a common population is negligible. We derive expressions for the expected values of commonly used measures of association, which confirm previous findings that the underlying mechanism of a genuine allele flip is variation in the haplotype frequencies and show further how this variation interacts with variation in the genetic effects to impact allele flipping. We show that for association testing at proxy SNPs, common in genome wide association studies, variation in haplotype frequencies must coincide with a reversal in the sign of linkage disequilibrium (LD) to trigger genuine allele flips. Using HapMap data and r, rather than r(2), to highlight previously unobserved effects, we show that unless genetic effects are large, variation in LD is unlikely to cause genuine allele flips in samples drawn from the same population. However, as populations diverge, it is an increasingly viable cause of a genuine allele flip for sufficiently large genetic effect and/or sample sizes. We conclude that evidence of variation in local patterns of LD, ancestral composition of study samples, and environmental exposures between study populations can provide compelling practical evidence in defense of a genuine allele flip. PMID- 20013944 TI - Motion control at the nanoscale. AB - Synthetic nanoscale motors represent a major step in the development of practical nanomachines. This Review summarizes recent progress towards controlling the movement of fuel-driven nanomotors and discusses the challenges and opportunities associated with the achievement of such nanoscale motion control. Regulating the movement of artificial nanomotors often follows nature's elegant and remarkable approach for motion control. Such on-demand control of the movement of artificial nanomotors is essential for performing various tasks and diverse applications. These applications require precise control of the nanomotor direction as well as temporal and spatial regulation of the motor speed. Different approaches for controlling the motion of catalytic nanomotors have been developed recently, including magnetic guidance, thermally driven acceleration, an electrochemical switch, and chemical stimuli (including control of the fuel concentration). Such ability to control the directionality of artificial nanomotors and to regulate their speed offers considerable promise for designing powerful nanomachines capable of operating independently and meeting a wide variety of future technological needs. PMID- 20013942 TI - Gene, region and pathway level analyses in whole-genome studies. AB - In the setting of genome-wide association studies, we propose a method for assigning a measure of significance to pre-defined sets of markers in the genome. The sets can be genes, conserved regions, or groups of genes such as pathways. Using the proposed methods and algorithms, evidence for association between a particular functional unit and a disease status can be obtained not just by the presence of a strong signal from a SNP within it, but also by the combination of several simultaneous weaker signals that are not strongly correlated. This approach has several advantages. First, moderately strong signals from different SNPs are combined to obtain a much stronger signal for the set, therefore increasing power. Second, in combination with methods that provide information on untyped markers, it leads to results that can be readily combined across studies and platforms that might use different SNPs. Third, the results are easy to interpret, since they refer to functional sets of markers that are likely to behave as a unit in their phenotypic effect. Finally, the availability of gene level P-values for association is the first step in developing methods that integrate information from pathways and networks with genome-wide association data, and these can lead to a better understanding of the complex traits genetic architecture. The power of the approach is investigated in simulated and real datasets. Novel Crohn's disease associations are found using the WTCCC data. PMID- 20013945 TI - Human spleen tyrosine kinase (Syk) recombinant expression systems for high throughput assays. AB - Spleen tyrosine kinase (Syk) is an important non-receptor tyrosine kinase and its aberrant regulation is associated with a variety of allergic disorders and autoimmune diseases. To identify small molecule inhibitors of Syk in high throughput assays, recombinant Syk protein is needed in bulk quantity. We studied the expression of recombinant human Syk in three heterologous systems: E. coli, baculovirus expression vector system (BEVS), and the cellular slime mold Dictyostelium discoideum (Dd). Syk activity was higher in the BEVS as compared to the Dd expression host, whereas in E. coli, no activity was observed under our assay conditions. Purified Syk kinase domain protein from BEVS showed concentration dependent inhibition with OXSI-2, a known Syk inhibitor. Molecular modeling and docking studies were performed to understand the binding mode and critical interactions of the inhibitor with catalytic domain of Syk. The BEVS generated Syk kinase domain showed stability upon multiple freeze-thaw cycles and exhibited significantly higher levels of tyrosine phosphorylation at pTyr(525)/Tyr(526) in the Syk activation loop. Based on our data, we conclude that BEVS is the ideal host to produce an active and stable enzyme, which can be successfully employed for screening of Syk inhibitors in a high-throughput system. PMID- 20013946 TI - From experimental setup to bioinformatics: an RNAi screening platform to identify host factors involved in HIV-1 replication. AB - RNA interference (RNAi) has emerged as a powerful technique for studying loss-of function phenotypes by specific down-regulation of gene expression, allowing the investigation of virus-host interactions by large-scale high-throughput RNAi screens. Here we present a robust and sensitive small interfering RNA screening platform consisting of an experimental setup, single-cell image and statistical analysis as well as bioinformatics. The workflow has been established to elucidate host gene functions exploited by viruses, monitoring both suppression and enhancement of viral replication simultaneously by fluorescence microscopy. The platform comprises a two-stage procedure in which potential host factors are first identified in a primary screen and afterwards re-tested in a validation screen to confirm true positive hits. Subsequent bioinformatics allows the identification of cellular genes participating in metabolic pathways and cellular networks utilised by viruses for efficient infection. Our workflow has been used to investigate host factor usage by the human immunodeficiency virus-1 (HIV-1), but can also be adapted to other viruses. Importantly, we expect that the description of the platform will guide further screening approaches for virus host interactions. The ViroQuant-CellNetworks RNAi Screening core facility is an integral part of the recently founded BioQuant centre for systems biology at the University of Heidelberg and will provide service to external users in the near future. PMID- 20013948 TI - On-the-fly targeted selection of labeled peptides in liquid chromatography/mass spectrometry based quantitative proteomics. PMID- 20013949 TI - Applications of electrospray laser desorption ionization mass spectrometry for document examination. AB - We have employed electrospray laser desorption ionization mass spectrometry (ELDI MS) to rapidly characterize certain classes of compounds--the inks within the characters made by inks and inkjet printer on regular paper and the chemical compounds within thermal papers. This ELDI-MS approach allowed the ink and paper samples to be distinguished in terms of their chemical compositions. Sample pretreatment was unnecessary and the documents were practically undamaged after examination. The ink chemicals on the documents were desorbed through laser irradiation (sampling spot area: <100 microm(2)); the desorbed molecules then entered an electrospray plume--prepared from an acidic methanol/water solution (50%)--where they became ionized through fusion or ion-molecule reactions with the charged solvent species and droplets in the plume. PMID- 20013950 TI - Identification of human calculi with time-of-flight secondary ion mass spectrometry. AB - Time-of-flight secondary ion mass spectrometry was used to study four human calculi and to compare the results with those from twelve commercially available urinary calculi minerals including three organic compounds (L-cystine, uric acid and sodium urate). Phase identification of calcium phosphate compounds was carried out by considering the relative ion abundances of [Ca(2)O](+) and [CaPO(2)](+). Deprotonated [M-H](-) and protonated [M+H](+) uric acid were detected and used for component recognition in pure uric acid and in the mixed samples of struvite, calcium oxalate and uric acid. Iodine related to the medical history of a patient was also detected. PMID- 20013951 TI - Reduction of bias in static closed chamber measurement of delta13C in soil CO2 efflux. AB - The (13)C/(12)C ratio of soil CO(2) efflux (delta(e)) is an important parameter in studies of ecosystem C dynamics, where the accuracy of estimated C flux rates depends on the measurement uncertainty of delta(e). The static closed chamber method is frequently used in the determination of delta(e), where the soil CO(2) efflux is accumulated in the headspace of a chamber placed on top of the soil surface. However, it has recently been shown that the estimate of delta(e) obtained by using this method could be significantly biased, which potentially diminish the usefulness of delta(e) for field applications. Here, analytical and numerical models were used to express the bias in delta(e) as mathematical functions of three system parameters: chamber height (H), chamber radius (R(c)), and soil air-filled porosity (theta). These expressions allow optimization of chamber size to yield a bias, which is at a level suitable for each particular application of the method. The numerical model was further used to quantify the effects on the delta(e) bias from (i) various designs for sealing of the chamber to ground, and (ii) inclusion of the commonly used purging step for reduction of the initial headspace CO(2) concentration. The present modeling work provided insights into the effects on the delta(e) bias from retardation and partial chamber bypass of the soil CO(2) efflux. The results presented here supported the continued use of the static closed chamber method for the determination of delta(e), with improved control of the bias component of its measurement uncertainty. PMID- 20013952 TI - Non-disturbing characterization of natural organic matter (NOM) contained in clay rock pore water by mass spectrometry using electrospray and atmospheric pressure chemical ionization modes. AB - We have investigated the composition of the mobile natural organic matter (NOM) present in Callovo-Oxfodian pore water using electrospray ionization mass spectrometry (ESI-MS), atmospheric pressure chemical ionization mass spectrometry (APCI-MS) and emission-excitation matrix (EEM) spectroscopy. The generation of knowledge of the composition, structure and size of mobile NOM is necessary if one wants to understand the interactions of these compounds with heavy metals/radionuclides, in the context of environmental studies, and particularly how the mobility of these trace elements is affected by mobile NOM. The proposed methodology is very sensitive in unambiguously identifying the in situ composition of dissolved NOM in water even at very low NOM concentration, due to innovative non-disturbing water sampling and ionization (ESI/APCI-MS) techniques. It was possible to analyze a quite exhaustive inventory of the small organic compounds of clay pore water without proceeding to any chemical treatment at naturally occurring concentration levels. The structural features observed were mainly acidic compounds and fatty acids as well as aldehydes and amino acids. PMID- 20013953 TI - Combining [13C6]-phenylisothiocyanate and the Edman degradation reaction: a possible breakthrough for absolute quantitative proteomics together with protein identification. AB - This manuscript describes the results of a preliminary experiment performed as 'proof of concept' of a novel approach to absolute quantitation of proteins without the use of standard proteins. Absolute quantitation remains a challenging issue in the proteomics field. Therefore, we propose a combination of [(13)C(6)] phenylisothiocyanate (PITC) and the Edman degradation reaction as a possible breakthrough. [(13)C(6)]-PITC was synthesized from [(13)C(6)]-aniline with O,O' di-2-pyridyl thiocarbonate to prepare [(13)C(6)]-phenylthiohydantoin (PTH)-amino acids as internal standards. Upon the Edman degradation reaction, it has been confirmed that a model protein, bovine serum albumin (BSA), releases the N terminal amino acid quantitatively as PTH-Asp. The standard curve of PTH-Asp against [(13)C(6)]-PTH-Asp showed good linearity (r(2) = 0.9977). BSA could be quantified as PTH-Asp using the standard curve. In addition, the residual des Asp(1)-BSA provided sufficient information for further protein identification. PMID- 20013954 TI - Role of PKCzeta translocation in the development of type 2 diabetes in rats following continuous glucose infusion. AB - AIM: We investigated the molecular mechanisms of hyperglycaemia-induced insulin resistance and type 2 diabetes in rats receiving a continuous glucose infusion (GI). METHODS: Female Wistar rats were infused with either 2.8 mol/L glucose or saline (2 mL/h) for durations varying from 0 to 15 days. Blood samples were analysed daily to determine glucose and insulin dynamics. Subsets of animals were sacrificed and soleus muscles were extracted for determination of protein expression, subcellular location, and activities of insulin-signalling proteins. RESULTS: Rats accommodated this systemic glucose oversupply and developed insulin resistance on day 5 (normoglycaemia/hyperinsulinaemia) and type 2 diabetes on day 15 (hyperglycaemia/normoinsulinaemia). The effect of GI on protein kinase Czeta (PKCzeta) activity was independent of changes in phosphatidylinositol 3-kinase activity, and occurred in parallel with an increase in PDK1 activity. Activated PKCzeta was mainly located in the cytosol after 5 days of GI that was coincident with the translocation of GLUT4 to the plasma membrane, and normoglycaemia. After 15 days of GI, PKCzeta translocated from the cytosol to the plasma membrane with a concomitant decrease in PDK1 activity. This caused an increase in the association between PKCzeta and PKB and a decrease in PDK1-PKB reactions at the plasma membrane, leading to reduced PKB activity. The activity of PKCzeta per se was also compromised. The PKCzeta and PKB activity reduction and the blunted insulin-stimulated GLUT4 translocation eventually led to hyperglycaemia and diabetes. CONCLUSION: Translocation of PKCzeta may play a central role in the development of type 2 diabetes. PMID- 20013955 TI - Adipokinetic hormone (Pyrap-AKH) enhances the effect of a pyrethroid insecticide against the firebug Pyrrhocoris apterus. AB - BACKGROUND: Adipokinetic hormones (AKHs) are insect neuropetides controlling stress situations including those elicited by insecticide treatment. The effect of Pyrap-AKH on the mortality of the firebug Pyrrhocoris apterus (L.) treated with the insecticide permethrin (Ambush 25 EC) was studied. RESULTS: Coinjection of 50 ng permethrin with 80 pmol Pyrap-AKH induced a significant 2.3-fold increase in bug mortality compared with the insecticide alone. The results were confirmed by topical coapplication of both agents (400 ng and 80 pmol respectively). Injections of 50 and 100 ng permethrin elicited a significant increase in the AKH level in CNS and the haemolymph. The results indicate an involvement of AKH in stress response to permethrin. The enhanced effect of insecticide by AKH treatments probably results from the stimulatory role in bug metabolism: carbon dioxide production was increased 3.5- and 2.5-fold respectively 1 and 3 h after permethrin treatment, and 4.3- and 3.4-fold after the permethrin plus AKH cotreatment, compared with the control. CONCLUSION: The elevation of metabolism could intensify the permethrin action by its faster penetration into tissues and by stimulation of biochemically active cells, and could be a reason for enhanced action of permethrin after its cotreatment with Pyrap-AKH. PMID- 20013956 TI - Analysis of phenolic and triterpenoid compounds in licorice and rat plasma by high-performance liquid chromatography diode-array detection, time-of-flight mass spectrometry and quadrupole ion trap mass spectrometry. AB - High-performance liquid chromatography with diode-array detection (HPLC/DAD), time-of-flight mass spectrometry (HPLC/TOFMS) and quadrupole ion trap mass spectrometry (HPLC/QITMS) were used for separation and identification of several compounds in licorice and rat plasma after oral administration of the herbal extract. Three phenolic compounds and one triterpenoid in licorice extract were unambiguously identified by comparing with the standard compounds. A formula database of known compounds in licorice was established, against which the other 42 compounds were identified effectively based on the accurate extract masses and formulae acquired by HPLC/TOFMS. In order to differentiate the isomers, tandem mass spectrometry was also used. The deduced fragmentation behaviors in QITMS were used to distinguish seven groups of isomers in licorice. By means of the three detectors, 46 compounds in licorice were identified. After oral administration of the extract, 25 compounds in rat plasma were detected and identified by comparing and contrasting the compounds measured in licorice with those in the plasma samples by HPLC/TOFMS. It is concluded that a rapid and effective method based on three analytical techniques was established, which is useful for identification of multiple compounds in licorice in vitro and in vivo. The result should be very useful for the quality control and curative mechanism study of licorice. PMID- 20013958 TI - Efficacy of interpersonal therapy-group format adapted to post-traumatic stress disorder: an open-label add-on trial. AB - BACKGROUND: Post-traumatic stress disorder (PTSD) is a highly prevalent condition, yet available treatments demonstrate only modest efficacy. Exposure therapies, considered by many to be the "gold-standard" therapy for PTSD, are poorly tolerated by many patients and show high attrition. We evaluated interpersonal therapy, in a group format, adapted to PTSD (IPT-G PTSD), as an adjunctive treatment for patients who failed to respond to conventional psychopharmacological treatment. METHODS: Research participants included 40 patients who sought treatment through a program on violence in the department of psychiatry of Federal University of Sao Paulo (UNIFESP). They had received conventional psychopharmacological treatment for at least 12 weeks and failed to have an adequate clinical response. After signing an informed consent, approved earlier by the UNIFESP Ethics Review Board, they received a semi-structured diagnostic interview (SCID-I), administered by a trained mental health worker, to confirm the presence of a PTSD diagnosis according to DSM-IV criteria. Other instruments were administered, and patients completed out self-report instruments at baseline, and endpoint to evaluate clinical outcomes. RESULTS: Thirty-three patients completed the trial, but all had at least one second outcome evaluation. There were significant improvements on all measures, with large effect sizes. CONCLUSIONS: IPT-G PTSD was effective not only in decreasing symptoms of PTSD, but also in decreasing symptoms of anxiety and depression. It led to significant improvements in social adjustment and quality of life. It was well tolerated and there were few dropouts. Our results are very preliminary; they need further confirmation through randomized controlled clinical trials. PMID- 20013957 TI - Clinical and subthreshold panic disorder. AB - BACKGROUND: Panic disorder (PD) is a highly prevalent and disabling condition, and subthreshold cases may be even more prevalent. This study examined psychiatric comorbidities, work functioning, and health-care utilization of individuals with subthreshold panic. The purpose of this study was to add to the accumulating evidence of significant comorbidity and disability associated with subthreshold PD. METHODS: Data are drawn from the Health Care for Communities study, a national household survey of the United States' adult, civilian, noninstitutionalized population (N=9585). Data assessing psychiatric disorders, employment and work productivity, and health-care utilization were collected. Seven categories of panic and subthreshold panic were created. RESULTS: The prevalence of clinical and subthreshold panic in the general population was 40%. Subthreshold panic was associated with increased odds of several comorbid disorders, including depression, dysthymia, psychosis, generalized anxiety disorder, bipolar disorder, and alcohol and drug use disorders. Subthreshold panic was also associated with greater likelihood of health-care service utilization but not with the intensity of mental health services. CONCLUSION: Psychiatric comorbidities and health-care utilization are common among patients with subthreshold panic. The relationship between varying degrees of panic symptomology and other mental health problems and indices of functioning impairment warrants further investigation. These results inform further research focusing on the course of subthreshold PD and its impact on quality of life. PMID- 20013959 TI - An adjunctive Management of Depression Program for difficult-to-treat depressed patients and their families. AB - BACKGROUND: The goal of this open-label feasibility trial was to test a short term, adjunctive intervention, the Management of Depression (MoD) Program, to determine if patients with difficult-to-treat forms of depression and their family members could learn to cope more effectively with their illness. METHODS: Nineteen patients meeting The Diagnostic and Statistical Manual IV criteria for major depressive disorder, dysthymia, or chronic/recurrent depression and their family members participated in an open-label study testing the efficacy of the MoD Program. The intervention consisted of nine sessions over 16 weeks, followed by an 8-month maintenance phase. Outcome measures focused on quality of life, psychological and family functioning, and level of depression. RESULTS: Fourteen patients and their family members improved significantly in psychosocial and family functioning, and depression severity (all P-values <.05) by the end of the 16-week intervention. There was also significant improvement in quality of life, psychosocial and family functioning, and depression scores (all P-values<.05) for the 10 patients who completed the maintenance phase. CONCLUSION: The MoD Program is a useful adjunctive intervention that helped patients and their family members deal more effectively with their persisting depression. The disease management approach improved the patient's perceived quality of life and functioning, reduced depressive symptoms, and improved perception of their family's functioning. PMID- 20013962 TI - Efficient aerobic Wacker oxidation of styrenes using palladium bis(isonitrile) catalysts. AB - The palladium-catalyzed aerobic oxidation of alkenes and especially styrenes (Wacker oxidation) by using chiral pseudo C(2)-symmetrical bis(isonitrile) ligands in the absence of further cocatalysts gives rise to methyl ketones in a highly chemoselective manner. The palladium bis(isonitrile) catalyst was characterized by NMR spectroscopy and X-ray structure analysis, revealing a dissymmetric coordination of palladium by the two isonitrile moieties. PMID- 20013961 TI - Influence of gold nanoparticles of varying size in improving the lipase activity within cationic reverse micelles. AB - Herein, we report the effect of gold nanoparticles (GNPs) in enhancing lipase activity in reverse micelles of cetyltrimethylammonium bromide (CTAB)/water/isooctane/n-hexanol. The size and concentration of the nanoparticles were varied and their specific roles were assessed in detail. An overall enhancement of activity was observed in the GNP-doped CTAB reverse micelles. The improvement in activity becomes more prominent with increasing concentration and size of the GNPs (0-52 microM and ca. 3-30 nm, respectively). The observed highest lipase activity (k(2)=1070+/-12 cm(3) g(-1) s(-1)) in GNP-doped CTAB reverse micelles ([GNP]: 52 microm, ca. 20 nm) is 2.5-fold higher than in CTAB reverse micelles without GNPs. Improvement in the lipase activity is only specific to the GNP-doped reverse micellar media, whereas GNP deactivates and structurally deforms the enzyme in aqueous media. The reason for this activation is probably due to the formation of larger-sized reverse micelles in which the GNP acts as a polar core and the surfactants aggregate around the nanoparticle ('GNP pool') instead of only water. Lipase at the augmented interface of the GNP doped reverse micelle showed improved activity because of enhancement in both the substrate and enzyme concentrations and increased flexibility in the lipase conformation. The extent of the activation is greater in the case of the larger sized GNPs. A correlation has been established between the activity of lipase and its secondary structure by using circular dichroism and FTIR spectroscopic analysis. The generalized influence of GNP is verified in the reverse micelles of another surfactant, namely, cetyltripropylammonium bromide (CTPAB). TEM, dynamic light scattering (DLS), and UV/Vis spectroscopic analysis were utilized to characterize the GNPs and the organized aggregates. For the first time, CTAB based reverse micelles have been found to be an excellent host for lipase simply by doping with appropriately sized GNPs. PMID- 20013960 TI - Research article: Antidepressant use among Asians in the United States. AB - OBJECTIVES: We examined the prevalence and predictors of past-year antidepressant use in a nationally representative sample of Asian Americans and non-Latino Whites. METHODS: Analyses of 12-month antidepressant medication use were based on data from the Collaborative Psychiatric Epidemiology Surveys that surveyed Asian (Chinese, Filipino, Vietnamese, and others; N=2,284) and non-Latino White (N=6,696) household residents ages 18 years and older in the 48 contiguous United States and Hawaii. RESULTS: Prevalence rates for 12-month antidepressant use for Asians with major depression ranged from 8.7% among Vietnamese to 17% among Chinese respondents. Compared to non-Latino Whites (32.4%), all Asians (10.9%) meeting criteria for 12-month depressive and anxiety disorders, but especially Filipinos (8.8%) were less likely to report past-year antidepressant use. CONCLUSIONS: We found disparities in past-year antidepressant use among all the examined major Asian groups meeting criteria for 12-month depressive and anxiety disorders. These disparities were not explained by mental health need or socioeconomic factors that enable access to care. PMID- 20013963 TI - Photophysical properties and OLED applications of phosphorescent platinum(II) Schiff base complexes. AB - The syntheses, crystal structures, and detailed investigations of the photophysical properties of phosphorescent platinum(II) Schiff base complexes are presented. All of these complexes exhibit intense absorption bands with lambda(max) in the range 417-546 nm, which are assigned to states of metal-to ligand charge-transfer ((1)MLCT) (1)[Pt(5d)-->pi*(Schiff base)] character mixed with (1)[lone pair(phenoxide)-->pi*(imine)] charge-transfer character. The platinum(II) Schiff base complexes are thermally stable, with decomposition temperatures up to 495 degrees C, and show emission lambda(max) at 541-649 nm in acetonitrile, with emission quantum yields up to 0.27. Measurements of the emission decay times in the temperature range from 130 to 1.5 K give total zero field splitting parameters of the emitting triplet state of 14-28 cm(-1). High performance yellow to red organic light-emitting devices (OLEDs) using these platinum(II) Schiff base complexes have been fabricated with the best efficiency up to 31 cd A(-1) and a device lifetime up to 77 000 h at 500 cd m(-2). PMID- 20013964 TI - Highly enantioselective synthesis of beta-heteroaryl-substituted dihydrochalcones through Friedel-Crafts alkylation of indoles and pyrrole. AB - A highly enantioselective Friedel-Crafts (F-C) alkylation of indoles and pyrrole with chalcone derivatives catalyzed by a chiral N,N'-dioxide-Sc(OTf)(3) complex has been developed that tolerates a wide range of substrates. The reaction proceeds in moderate to excellent yields and high enantioselectivities (85-92 % enantiomeric excess) using 2 mol % (for indole) or 0.5 mol % (for pyrrole) catalyst loading, which showed the potential value of the catalyst system. Meanwhile, a strong positive nonlinear effect was observed. On the basis of the experimental results and previous reports, a possible working model is proposed to explain the origin of the activation and asymmetric induction. PMID- 20013965 TI - Pyrolysis of a three-dimensional Mn(II)/Mn(III) network to give a multifunctional porous manganese oxide material. PMID- 20013966 TI - Evolution of helical mesostructures. AB - An intriguing evolution from a simple internal helix to a hierarchical helical (HH) mesostructure with both internal and external helices or a complicated screwlike and concentric circular (CC) mesostructure is successfully observed. The complicated helical structures are determined by TEM studies and 3D electron tomography. We demonstrate a topological helix-coil transition between the internal and external helices to reveal the origin of the HH mesostructure and the relationship between the straight helical and HH rods. Moreover, the boundary condition of the helix-coil transition is clarified to explain in detail the formation of complex helical structures, such as the screwlike mesostructure. It is proposed that the final structural characteristics are determined exactly by the balance between the decrease in the surface free energy and the maintenance of the hexagonal packing in one individual rod, which explains the formation of unusual CC, HH, and screwlike morphologies in one pot. Our success has opened new opportunities in the characterization of complex porous architectures, thus paving a way to remarkable advances in the fields of synthesis, understanding, and application of novel porous materials. PMID- 20013967 TI - Layered double hydroxide-supported L-methionine-catalyzed chemoselective O methylation of phenols and esterification of carboxylic acids with dimethyl carbonate: a "green" protocol. PMID- 20013968 TI - Chiral auxiliaries as emerging tools for the asymmetric synthesis of octahedral metal complexes. AB - New methods for the stereocontrolled synthesis of octahedral metal complexes are needed in order to fully exploit the stereochemical richness of the octahedron in the fields of catalysis, materials sciences, and life sciences. Whereas a large body of work exists regarding the diastereoselective coordination chemistry with chiral ligands, such efforts are restricted to certain carefully designed chiral ligands which remain in the coordination sphere. The emerging strategy of chiral auxiliary-mediated asymmetric synthesis holds promise to solve the problem of controlling relative and absolute configuration in octahedral metal complexes in a general fashion, thus hopefully in the future providing access to any desired optical active octahedral metal complex without the need for chiral separations. This short review will summarize reported examples of chiral auxiliaries applied to the asymmetric synthesis of octahedral metal complexes. PMID- 20013969 TI - Postsynthetic modification of peptides: chemoselective C-arylation of tryptophan residues. PMID- 20013970 TI - Cyclometallation and hydrogen/deuterium exchange reactions of an arylphosphine ligand upon coordination to {Ir(eta(5)-C(5)Me(5))}. PMID- 20013971 TI - [12]Annulynes from 1,5-hexadiyne and potassium tert-butoxide? Franz sondheimer's hexadienynes! PMID- 20013972 TI - Direct STD NMR identification of beta-galactosidase inhibitors from a virtual dynamic hemithioacetal system. PMID- 20013973 TI - Combined rhodium-catalyzed carbon-hydrogen activation and beta-carbon elimination to access eight-membered rings. PMID- 20013974 TI - Nitrated benzyne derivatives of La@C(82): addition of NO(2) and its positional directing effect on the subsequent addition of benzynes. PMID- 20013975 TI - Template-directed synthesis of nanoplasmonic arrays by intracrystalline metalization of cross-linked lysozyme crystals. PMID- 20013976 TI - Strain and huckel aromaticity: driving forces for a promising new generation of electron acceptors in organic electronics. PMID- 20013977 TI - Lucky seven: characterization of stable T-shaped copper(II) complexes of [32]heptaphyrins. PMID- 20013978 TI - Water-vapor-induced reversible switching of electronic states in an MMX-type chain complex with retention of single crystallinity. PMID- 20013979 TI - Efficient synthesis of aryl fluorides. PMID- 20013980 TI - Biomimetic formation of 2-tropolones by dioxygenase-catalysed ring expansion of substituted 2,4-cyclohexadienones. AB - Substituted 2-tropolone natural products are found in plants and fungi. Their biosynthesis is thought to occur by ring expansion from a cyclohexadienone precursor, but this reaction has not previously been demonstrated experimentally. Treatment of 6-hydroxy-6-hydroxymethylcyclohexa-2,4-dienone with the non-haem iron(II)-dependent extradiol catechol dioxygenase MhpB from Escherichia coli results in the formation of the 2-tropolone ring-expansion product through a pinacol-type rearrangement. Three further substituted cyclohexa-2,4-dienone analogues were prepared, and treatment of each analogue was found to give the substituted 2-tropolone ring-expansion product. This ring expansion could also be effected nonenzymatically by treatment with 1,4,7-triazacyclononane and FeCl(2). This is a novel transformation for non-haem iron-dependent enzymes, and this is the first experimental demonstration of the proposed ring-expansion reaction in tropolone biosynthesis. PMID- 20013981 TI - Protein incorporation in giant lipid vesicles under physiological conditions. PMID- 20013982 TI - Phosphopantetheinylation and specificity of acyl carrier proteins in the mupirocin biosynthetic cluster. AB - Acyl carrier proteins are vital for the biosynthesis of fatty acids and polyketides. The mupirocin biosynthetic cluster of Pseudomonas fluorescens encodes eleven type I ACPs embedded in its multifunctional polyketide synthase (PKS) proteins plus five predicted type II ACPs (mAcpA-E) that are known to be essential for mupirocin biosynthesis by deletion and complementation analysis. MupN is a putative Sfp-type phosphopantetheinyl transferase. Overexpression of three type I and three type II mupirocin ACPs in Escherichia coli, with or without mupN, followed by mass spectroscopy revealed that MupN can modify both mupirocin type I and type II ACPs to their holo-form. The endogenous phosphopantetheinyl transferase of E. coli modified mAcpA but not mAcpC or D. Overexpression of the type II ACPs in macp deletion mutants of the mupirocin producer P. fluorescens 10586 showed that they cannot substitute for each other while hybrids between mAcpA and mAcpB indicated that, at least for mAcpB, the C terminal domain determines functional specificity. Amino acid alignments identified mACPs A and D as having C-terminal extensions. Mutation of these regions generated defective ACPs, the activity of which could be restored by overexpression of the macp genes on separate plasmids. PMID- 20013983 TI - Coulomb physics in spin ice: from magnetic monopoles to magnetic currents. PMID- 20013984 TI - Alignment of carbon nanotubes in thermotropic and lyotropic liquid crystals. AB - The organization of carbon nanotubes is an important but challenging issue. This Minireview focuses on a novel approach based on the use of liquid crystals as host for nanotube dispersion and ordering. With these self-organizing fluids it is possible to control the nanotube orientation on a macroscopic scale, as the orientational order of the liquid crystal, the direction of which can easily be selected and dynamically modulated, is transferred onto embedded carbon nanotubes. Both main classes of liquid crystals, thermotropics and lyotropics, have been used successfully. This reveals an aligning mechanism resulting from the liquid crystallinity per se, independent of specific molecular structures, although these can affect the degree of order and the quality of the tube dispersion. The enormous variety of systems that form liquid crystalline phases provides additional strength and potentialities to the liquid-crystal-based route towards nanotube alignment. The exploration of this new use of liquid crystals has just started, hence we can expect exciting developments in the years to come. PMID- 20013985 TI - Banking a hemiabdominal DIEP flap: oncologic indications. PMID- 20013986 TI - Symptoms of traumatic stress in mothers of children victims of a motor vehicle accident. AB - BACKGROUND: Motor vehicle accidents (MVAs) are the main cause of Posttraumatic stress disorder (PTSD) in industrialized countries. This includes the frequently occurring but understudied situation of parents learning that their children were injured. However, unlike in other types of trauma survivors, little is known about the predictors of PTSD symptoms in mothers whose child has suffered an MVA. METHODS: A group of 72 mothers and 28 fathers were prospectively assessed for peritraumatic distress, peritraumatic dissociation, and PTSD symptoms 1 and 5 weeks after their child had suffered an MVA. RESULTS: Levels of peritraumatic distress and dissociation were comparable to other trauma victims, 18% of the mothers were considered to be suffering from probable PTSD. In mothers, significant positive correlations were found between PTSD symptoms and peritraumatic distress (r=.34) and dissociation (r=.37), whereas mothers' PTSD symptoms were associated with decreased peritraumatic dissociation in fathers (r= .37). Even after controlling for covictim/witness status, peritraumatic distress was a predictor of mothers' PTSD symptoms, explaining 14% of the variance. CONCLUSIONS: Peritraumatic response and PTSD symptoms should be routinely assessed among parents whose child has experienced a traumatic event. PMID- 20013987 TI - Anxiety does not predict response to duloxetine in major depression: results of a pooled analysis of individual patient data from 11 placebo-controlled trials. AB - BACKGROUND: Uncontrolled antidepressant trials suggest that anxious patients with major depressive disorder (MDD) are less responsive to antidepressant treatment than less anxious patients. The objective of this study is to determine whether specific antidepressant effects, estimated by drug-placebo differences, are reduced in anxious depression during treatment of MDD with duloxetine. METHODS: This is a retrospective secondary pooled analysis of all placebo-controlled trials of duloxetine at therapeutic doses conducted by the sponsor in outpatients with nonpsychotic unipolar MDD, using the Hamilton Depression Rating Scale (HAMD). Anxious depression was defined by > or =7 on the anxiety/somatization factor of the HAMD. Response was defined as > or =50% improvement from baseline to endpoint on the HAMD. Remission was defined as an endpoint HAMD< or =7. Analyses were performed in the intent-to-treat sample with at least one post treatment rating. RESULTS: Eleven trials included 2,841 patients of whom 1,326 were classified as anxious and 1,515 as nonanxious. Change on the HAMD was greater with duloxetine than placebo in both anxious (9.91 versus 7.55, P<.001) and nonanxious (6.65 versus 5.23, P<.001) patients. Level of anxiety had no effect on the drug-placebo differences. Response and remission rates were significantly greater in duloxetine than placebo-treated patients and drug placebo differences were unaffected by anxious status. Use of HAMD items psychic and somatic anxiety to define anxious subgroups had similar outcomes. CONCLUSIONS: Duloxetine was more effective than placebo in achieving response and remission in both anxious and nonanxious patients. Anxious status did not affect the magnitude of the drug effect. PMID- 20013988 TI - Inhibitors of cathepsins K and S identified using the DynaMAD virtual screening algorithm. PMID- 20013989 TI - Myrcene as a natural base chemical in sustainable chemistry: a critical review. AB - Currently, a shift towards chemical products derived from renewable, biological feedstocks is observed more and more. However, substantial differences with traditional feedstocks, such as their "hyperfunctionalization," ethical problems caused by competition with foods, and problems with a constant qualitative/quantitative availability of the natural products, occasionally complicate the large-scale market entry of renewable resources. In this context the vast family of terpenes is often not taken into consideration, although the terpenes have been known for hundreds of years as components of essential oils obtained from leaves, flowers, and fruits of many plants. The simple acyclic monoterpenes, particularly the industrially available myrcene, provide a classical chemistry similar to unsaturated hydrocarbons already known from oil and gas. Hence, this Review is aimed at reviving myrcene as a renewable compound suitable for sustainable chemistry in the area of fine chemicals. The versatility of the unsaturated C(10)-hydrocarbon myrcene, leading to products with several different areas of application, is pointed out. PMID- 20013990 TI - Voltammetry in room temperature ionic liquids: comparisons and contrasts with conventional electrochemical solvents. AB - The recent literature is surveyed to explore the nature of voltammetry in room temperature ionic liquids. The extent of similarities with conventional electrochemical solvents is reported and some surprising differences are noted. PMID- 20013991 TI - Multianalyte electrochemical biosensor based on aptamer- and nanoparticle integrated bio-barcode amplification. AB - In the present work, a signal-on electrochemical sensing strategy for the simultaneous detection of adenosine and thrombin is developed based on switching structures of aptamers. An Au electrode as the sensing surface is modified with two kinds of thiolated capture probes complementary to the linker DNA that contains either an adenosine aptamer or thrombin aptamer. The capture probes hybridize with their corresponding linker DNA, which has prehybridized with the reporter DNA loaded onto the gold nanoparticles (AuNPs). The AuNP contained two kinds of bio-barcode DNA: one is complementary to the linker DNA (reporter), whereas the other is not (signal) and is tagged with different metal sulfide nanoparticles. Thus a "sandwich-type" sensing interface is fabricated for adenosine and thrombin. With the introduction of adenosine and thrombin, the aptamer parts bind with their targets and fold to form the complex structures. As a result, the bio-barcoded AuNPs are released into solution. The metal sulfide nanoparticles are measured by anodic stripping voltammetry (ASV), and the concentrations of adenosine and thrombin are proportional to the signal of either metal ion. With the dual amplification of the bio-barcoded AuNP and the preconcentration of metal ions through ASV technology, detection limits as low as 6.6 x 10(-12) M for adenosine and 1.0 x 10(-12) M for thrombin are achieved. The sensor exhibits excellent selectivity and detectability in biological samples. PMID- 20013992 TI - Formation of alpha-lithio siloles from silylated 1,4-dilithio-1,3-butadienes: mechanism and applications. AB - A full account of a useful transformation from silylated 1,4-dilithio-1,3 butadienes to alpha-lithio siloles is described. These lithio siloles formed by this procedure are general, in terms of substitution patterns and synthetic methods, affording diversified silole derivatives. Notably, some structurally complex molecules, such as bridged bis-silole compounds, have been synthesized easily and successfully by applying our protocol. The structure of the alpha lithio silole, which adopts a dimeric fashion through two lithium bridges, was confirmed by X-ray analysis. Furthermore, a possible mechanism of the skeleton rearrangements via E/Z isomerization of 1-silyl-1-lithio alkene and nucleophilic attack on silicon is proposed, and is also proved by experimental investigations. PMID- 20013993 TI - Redox multifunctionality in a series of Pt(II) dithiolene complexes of a tetrathiafulvalene-based diphosphine ligand. AB - The redox-active and chelating diphosphine, 3,4-dimethyl-3',4' bis(diphenylphosphino)-tetrathiafulvalene, denoted as P2, is engaged in a series of platinum complexes, [(P2)Pt(dithiolene)], with different dithiolate ligands, such as 1,2-benzenedithiolate (bdt), 1,3-dithiole-2-thione-4,5-dithiolate (dmit), and 5,6-dihydro-1,4-dithiin-2,3-dithiolate (dddt). The complexes are structurally characterized by X-ray diffraction, together with a model compound derived from bis(diphenylphosphino)ethane, namely, [(dppe)Pt(dddt)]. Four successive reversible electron-transfer processes are found for the [(P2)Pt(dddt)] complex, associated with the two covalently linked but electronically uncoupled electrophores, that is, the TTF core and the platinum dithiolene moiety. The assignments of the different redox processes to either one or the other electrophore is made thanks to the electrochemical properties of the model compound [(dppe)Pt(dddt)] lacking the TTF redox core, and with the help of theoretical calculations (DFT) to understand the nature and energy of the frontier orbitals of the [(P2)Pt(dithiolene)] complexes in their different oxidation states. The first oxidation of the highly electron-rich [(P2)Pt(dddt)] complex can be unambiguously assigned to the redox process affecting the Pt(dddt) moiety rather than the TTF core, a rare example in the coordination chemistry of tetrathiafulvalenes acting as ligands. PMID- 20013994 TI - Synthesis of N-arylated 1,2-dihydroheteroaromatics through the three-component reaction of arynes with N-heteroaromatics and terminal alkynes or ketones. AB - The reaction of benzynes with N-heteroaromatics including quinolines, isoquinolines, and pyridines and various terminal alkynes or ketones with an alpha-hydrogen in the presence of KF and 18-crown-6 in THF at room temperature for 8 h gave various N-arylated 1,2-dihydroheteroaromatics in good to moderate yields. Some of these product structures are found in various naturally occurring and biologically active heterocyclic compounds. The reaction involves an unusual multiple construction of new C--C, C--N, and C--H bonds and the cleavage of a C- H bond in one pot. It is likely that the three-component coupling proceeds through the nucleophilic addition of quinoline to benzyne, which generates a zwitterionic species. The latter then attracts a proton from terminal alkyne (or ketone) to generate an N-arylated quinolinium cation and an acetylide anion. Further reaction of these two ions provides the final substituted 1,2 dihydroquinolines. In the reaction, the terminal alkyne acts first as a proton donor and then as a nucleophile. The application of a three-component coupling reaction product, 1,2-dihydro-2-pyridinyl alkyne in a stereospecific [4+2] Diels Alder cycloaddition reaction with N-phenyl maleimide to give an isoquinuclidine derivative, an important core present in various natural products, is demonstrated. PMID- 20013995 TI - Synthesis and characterization of water-soluble conjugated glycopolymer for fluorescent sensing of concanavalin A. AB - A neutral polyfluorene derivative that contains 20 mol% 2,1,3-benzothiadiazole (BT) is synthesized by Suzuki cross-coupling polymerization. A cationic conjugated polymer A and an alpha-mannose-bearing polymer B are subsequently obtained through different post-polymerization methods. As a result of the charged pendant groups or sugar-bearing groups attached to the polymer side chains, both A and B show good water-solubility. The titration of Concanavalin A (Con A) into polymer aqueous solution leads to different fluorescent responses for polymers A and B. Polymer A does not show any obvious fluorescence change upon interaction with Con A, whereas polymer B shows fluorescence increase in BT emission intensity when Con A is added. This is because of the specific interaction between alpha-mannose and Con A, which induces polymer aggregation, and then facilitates energy transfer from the phenylene-fluorene segments to the BT units. A practical calibration curve ranging from 1 nM to 250 nM is obtained by correlating the changes in BT emission intensity with Con A concentration. The advantage of polymer B-based Con A macromolecular probe is that it shows signal increase upon Con A recognition, which is significantly different from other conjugated polymer-based fluorescence quenching assays. PMID- 20013996 TI - An air stable moisture resistant titanium triflate complex as a Lewis acid catalyst for C-C bond forming reactions. AB - An air and moisture stable C(3)-symmetric titanium(IV) triflate, supported by a tripodal amine-(tris-phenolate) ligand, has been synthesized and characterized by X-ray crystallography and shown to be a good Lewis acid catalyst for a range of aza-Diels-Alder, Diels-Alder, syn aldol, allylation, and alkylation reactions. PMID- 20013997 TI - Phase-transfer-catalyzed asymmetric alkylation of alpha-benzoyloxy-beta-keto esters: stereoselective construction of congested 2,3-dihydroxycarboxylic acid esters. AB - Highly enantioselective phase-transfer-catalyzed alkylation of tert-butyl 2 benzoyloxy-3-oxobutanoate was realized by the use of an N-spiro chiral quaternary ammonium salt, as a complementary approach to the asymmetric hydroxylation of alpha-alkyl-beta-keto esters. The synthetic utility of the alkylated compounds is highlighted by the diastereoselective reduction and alkylation of the remaining ketone moiety to give various enantiomerically enriched congested 2,3 dihydroxycarboxylic acid esters. PMID- 20013998 TI - Rh(II)-catalyzed reaction of alpha-diazocarbonyl compounds bearing beta trichloroacetylamino substituent: C-H insertion versus 1,2-H shift. AB - The Rh(II)-carbene reaction is dramatically affected by the neighboring substituents. If the neighboring substituent is an OH group, a1,2-H shift is the exclusive pathway. If it is an OAc group, a 1,2-acetoxy migration is observed. If it is p-toluenesulfonyl group, 1,3 and 1,5-C-H insertion are the major pathways, and the 1,2-H shift is completely suppressed. If the adjacent substituent is a trichloroacetyl amino group, 1,5-C-H insertion competes with the 1,2-hydride shift, and no 1,3-C-H insertion can be observed. Both electronic and steric factors are responsible for the switching of the Rh(II)-carbene reaction pathway. The highly stereoselective 1,5-C-H insertions in Rh(II)-catalyzed reaction of alpha-diazocarbonyl compounds, bearing beta-trichloroacetylamino substituent, can be utilized as a novel way to synthesize five-membered cyclic beta-amino acid derivatives. PMID- 20013999 TI - Tautomycin's interactions with protein phosphatase 1. AB - It has been a long journey since tautomycin (TTM) was isolated in 1987 and the discovery that it inhibited protein phosphatase 1 (PP1) more strongly than PP2A until finally the cocrystal structure of TTM and PP1 was presented early in 2009. The fact that TTM shows preference to inhibit PP1 over PP2A makes this compound unique among the known PP1 and PP2A inhibitors. A number of groups were involved in work aiming to unravel TTM's interactions with PP1 and by doing so hoping to disentangle the secrets as to why TTM is a better inhibitor of PP1 than PP2A. This Focus Review looks back at the work conducted with TTM in order to establish its point of interaction with PP1 prior to X-ray structure. Finally the conclusions before the X-ray structure are compared with the real situation. PMID- 20014000 TI - Flexible and transparent silicon nanoparticle/polymer composites with stable luminescence. PMID- 20014001 TI - Cyclometallated iridium complexes as sensitizers for dye-sensitized solar cells. PMID- 20014002 TI - Cu(I)/L-proline-catalyzed coupling of substituted 3-iodoprop-2-en-1-ols with 1 alkynes and subsequent cyclization: a convenient approach for synthesizing polysubstituted furans. PMID- 20014003 TI - Platelet graphite nanofibers for electrochemical sensing and biosensing: the influence of graphene sheet orientation. AB - Here, we demonstrate that platelet graphite nanofibers (PGNFs) exhibit fast heterogeneous electron-transfer rates for a wide variety of compounds such as FeCl(3), ferrocyanide, dopamine, uric acid, ascorbic acid, and the reduced form of beta-nicotinamide adenine dinucleotide. The electrochemical properties of PGNFs are superior to those of multiwalled carbon nanotubes (MWCNTs) or graphite microparticles (GMPs). Transmission electron microscopy and Raman spectroscopy reveal that this arises from the unique graphene sheet orientation of such platelet nanofibers, which accounts for their unparalleled high ratio of graphene edge planes versus basal planes. PMID- 20014004 TI - What's in a name? Qualitative description revisited. AB - "Whatever Happened to Qualitative Description?" (Sandelowski, 2000) was written to critique the prevailing tendency in qualitative health research to claim the use of methods that were not actually used and to clarify a methodological approach rarely identified as a distinctive method. The article has generated several misconceptions, most notably that qualitative description requires no interpretation of data. At the root of these misconceptions is the persistent challenge of defining qualitative research methods. Qualitative description is a "distributed residual category" (Bowker & Star, 2000). Cambridge, MA: The MIT Press) in the classification of these methods. Its value lies not only in the knowledge its use can produce, but also as a vehicle for presenting and treating research methods as living entities that resist simple classification. PMID- 20014006 TI - Functional organization of mammalian mitochondrial DNA in nucleoids: history, recent developments, and future challenges. AB - Various proteins involved in replication, repair, and the structural organization of mitochondrial DNA (mtDNA) have been characterized in detail over the past 25 or so years. In addition, in recent years, many proteins were identified with a role in the dynamics of the mitochondrial network. Using advanced imaging and an increasing number of cytological techniques, we have begun to realize that an important aspect to mtDNA maintenance, in both health and disease, is its organization within the dynamic mitochondrial network in discrete protein-DNA complexes usually termed nucleoids. Here, I review recent developments in the study of nucleoid dynamics and proteins. I will discuss the implications of the organization of mtDNA in nucleoids in light of DNA replication, repair, gene expression, segregation, and inheritance. PMID- 20014005 TI - PELP1: A novel therapeutic target for hormonal cancers. AB - Recent studies implicate that the estrogen receptor (ER) coregulator proline-, glutamic acid-, and leucine-rich protein (PELP) 1 as playing critical roles in ER genomic, ER-nongenomic, and ER-signaling cross talk with growth factor signaling pathways. PELP1 expression is deregulated in hormonal cancers and recent studies further elucidated the molecular mechanisms by which PELP1 regulates hormone therapy response. Although PELP1 is important for normal functions of the ER, the possibility to target ER-PELP1 axis appears to be an effective strategy for preventing hormonal carcinogenesis and therapy resistance. Thus, PELP1 may be useful as prognostic marker for hormonal cancers and PELP1 signaling may be useful to generate targeted therapeutics to overcome hormonal therapy resistance. PMID- 20014007 TI - Early life stage and genetic toxicity of stannous chloride on zebrafish embryos and adults: toxic effects of tin on zebrafish. AB - Humans are exposed to stannous chloride (SnCl(2)), known as tin chloride, present in packaged food, soft drinks, biocides, dentifrices, etc. Health effects in children exposed to tin and tin compounds have not been investigated yet. Therefore, we evaluated the possible teratogenic effects and genotoxic of SnCl(2) in zebrafish (Danio rerio) adults and their embryos. In the embryo-larval study, SnCl(2) showed embryo toxicity and developmental delay after exposure to the various concentrations of 10-250 MUM for 120 h. Teratogenic effects including morphological malformations of the embryos and larvae were observed. The embryos exposed to 100 MUM displayed tail deformation at 28 hpf and the larvae exposed to 50 MUM showed reduced body growth, smaller head and eyes, bent trunk, mild pericardial edema, and smaller caudal fin at 96 hpf. The results of the teratological study show that SnCl(2) induced a significant decrease in the number of living embryos and larvae. Regarding the chromosome analysis, SnCl(2) induced a dose-dependent increase in the micronucleus (MN) frequency in peripheral erythrocytes of adult zebrafish. In blood cells, the 25 MUM dose of SnCl(2) caused a nonsignificant increase in the total chromosomal aberrations, but the high doses significantly increased the total number of chromosomal aberrations compared with the control groups. Overall, the results clearly indicate that SnCl(2) is teratogenic and genotoxic to zebrafish. PMID- 20014008 TI - Hsp70/Hsp90 co-chaperones are required for efficient Hsp104-mediated elimination of the yeast [PSI(+)] prion but not for prion propagation. AB - The continued propagation of the yeast [PSI(+)] prion requires the molecular chaperone Hsp104 yet in cells engineered to overexpress Hsp104; prion propagation is impaired leading to the rapid appearance of prion-free [psi(-)] cells. The underlying mechanism of prion loss in such cells is unknown but is assumed to be due to the complete dissolution of the prion aggregates by the ATP-dependent disaggregase activity of this chaperone. To further explore the mechanism, we have sought to identify cellular factors required for prion loss in such cells. Sti1p and Cpr7p are co-chaperones that modulate the activity of Hsp70/Ssa and Hsp90 chaperones and bind to the C-terminus of Hsp104. Neither Sti1p nor Cpr7p is necessary for prion propagation but we show that deletion of the STI1 and CPR7 genes leads to a significant reduction in the generation of [psi(-)] cells by Hsp104 overexpression. Deletion of the STI1 and CPR7 genes does not modify the elimination of [PSI(+)] by guanidine hydrochloride, which inhibits the ATPase activity of Hsp104 but does block elimination of [PSI(+)] by overexpression of either an ATPase-defective mutant of Hsp104 (hsp104(K218T/K620T)) or a 'trap' mutant Hsp104 (hsp104(E285Q/E687Q)) that can bind its substrate but can not release it. These results provide support for the hypothesis that [PSI(+)] elimination by Hsp104 overexpression is not simply a consequence of complete dissolution of the prion aggregates but rather is through a mechanism distinct from the remodelling activity of Hsp104. PMID- 20014009 TI - Intracerebral transplantation of bone marrow-derived mesenchymal stem cells reduces amyloid-beta deposition and rescues memory deficits in Alzheimer's disease mice by modulation of immune responses. AB - Alzheimer's disease (AD) is characterized by the deposition of amyloid-beta peptide (Abeta) and the formation of neurofibrillary tangles. Transplantation of bone marrow-derived mesenchymal stem cells (BM-MSCs) has been suggested as a potential therapeutic approach to prevent various neurodegenerative disorders, including AD. However, the actual therapeutic impact of BM-MSCs and their mechanism of action in AD have not yet been ascertained. The aim of this study was therefore to evaluate the therapeutic effect of BM-MSC transplantation on the neuropathology and memory deficits in amyloid precursor protein (APP) and presenilin one (PS1) double-transgenic mice. Here we show that intracerebral transplantation of BM-MSCs into APP/PS1 mice significantly reduced amyloid beta peptide (Abeta) deposition. Interestingly, these effects were associated with restoration of defective microglial function, as evidenced by increased Abeta degrading factors, decreased inflammatory responses, and elevation of alternatively activated microglial markers. Furthermore, APP/PS1 mice treated with BM-MSCs had decreased tau hyperphosphorylation and improved cognitive function. In conclusion, BM-MSCs can modulate immune/inflammatory responses in AD mice, ameliorate their pathophysiology, and improve the cognitive decline associated with Abeta deposits. These results demonstrate that BM-MSCs are a potential new therapeutic agent for AD. PMID- 20014010 TI - KMT1E mediated H3K9 methylation is required for the maintenance of embryonic stem cells by repressing trophectoderm differentiation. AB - Dynamic regulation of histone methylation by methyltransferases and demethylases plays a central role in regulating the fate of embryonic stem (ES) cells. The histone H3K9 methyltransferase KMT1E, formerly known as ESET or Setdb1, is essential to embryonic development as the ablation of the Setdb1 gene results in peri-implantation lethality and prevents the propagation of ES cells. However, Setdb1-null blastocysts do not display global changes in H3K9 methylation or DNA methylation, arguing against a genome-wide defect. Here we show that conditional deletion of the Setdb1 gene in ES cells results in the upregulation of lineage differentiation markers, especially trophectoderm-specific factors, similar to effects observed upon loss of Oct3/4 expression in ES cells. We demonstrate that KMT1E deficiency in ES cells leads to a decrease in histone H3K9 methylation at and derepression of trophoblast-associated genes such as Cdx2. Furthermore, we find genes that are derepressed upon Setdb1 deletion to overlap with known targets of polycomb mediated repression, suggesting that KMT1E mediated H3K9 methylation acts in concert with polycomb controlled H3K27 methylation. Our studies thus demonstrate an essential role for KMT1E in the control of developmentally regulated gene expression programs in ES cells. PMID- 20014011 TI - Social dynamics and individual plasticity of infant care behavior in cooperatively breeding cotton-top tamarins. AB - Individual variation in infant caretaking behavior is prevalent among marmoset and tamarin monkeys. Although most group members participate in infant care, the timing and amount provided differs greatly. In this study, we quantified general trends in infant carrying behavior by using a longitudinal database that included 11 years of instantaneous scan observations following 80 births of cotton-top tamarins. Using detailed focal observations on a subset of the same families (10 births) we identified influences that affected expression of infant care at the group and individual levels. Fathers were the primary carriers and paternal carry time gradually decreased with increasing infant age. Paternal carry time also decreased significantly with an increasing number of older sibling helpers. Most fathers began to carry on the first day postpartum. However, we report circumstances in which fathers delayed carrying until almost a month postpartum. Fathers retrieved infants the most, although adult brothers' rates of retrievals peaked and surpassed fathers' rates during week 4 postpartum. Fathers delayed rejection of infants until week 4, whereas mothers rejected infants immediately and throughout the eight weeks. Nonetheless, infants climbed onto their mothers more than onto any other family member. Mothers showed a high initial investment in carrying during the first two weeks, decreasing quickly thereafter. Maternal contributions to infant carrying remained low and relatively consistent regardless of group size. However, mothers dramatically increased their infant carrying behavior in families in which fathers were absent. Older siblings cared for infants more than did younger siblings, and brothers retrieved and carried infants more than did sisters. Individual expression of infant care changed to accommodate infant needs and changed according to varying social dynamics and circumstances across litters. PMID- 20014012 TI - Conflict and reconciliation behavior trends of the bottlenose dolphin (Tursiops truncatus). AB - Wild bottlenose dolphin (Tursiops truncatus) populations display societal structures characterized by numerous and frequent changes in group composition, complex social relationships, and high levels of cooperation, attributes also observed in human and nonhuman primate cultures. Maintaining social relationships under such elemental conditions can frequently create conflict--and the opportunity for reconciliation--among group members. The conflict and reconciliation behavior patterns of the chimpanzee (Pan troglodytes) have been studied extensively; trends are well-documented in the wild and in captivity. Apparent cultural similarities have prompted several analogous studies on wild and captive bottlenose dolphins. This research attempted to corroborate previous efforts by analyzing the social behavior of seven captive bottlenose dolphins to determine the effects of sex and age on the frequency of conflict and reconciliation, and to investigate the incidence of consolatory behavior within the group. A total of 3,428 interactions involving focal animals, 414 of which were conflict episodes, were documented during 261 hr of videotaped observations. Although the sample size precluded meaningful statistical evaluation of the influence of sex on conflict and reconciliation, participant age was a significant determinant of conflict frequency. Conversely, age did not impact frequency of reconciliation, which only occurred after 18% of all conflict interactions. Little to no definitive evidence of consolation was apparent within the study group. While results partially support the findings of previous dolphin reconciliation research, extensive behavioral studies of wild populations should be conducted before generating broad comparisons between human terms and nonhuman behavioral interactions. PMID- 20014013 TI - Special issue--the care and welfare of elephants in AZA institutions. PMID- 20014014 TI - New ultrasound-based imaging technologies are claimed to avoid unnecessary breast biopsies, but what is an "unnecessary" image-guided needle biopsy of the breast? PMID- 20014015 TI - Prenatal ultrasound and magnetic resonance imaging depiction of a small sublingual ranula. AB - Prenatal diagnosis of a congenital ranula has rarely been reported. We describe the case of a small ranula depicted on prenatal sonogram and magnetic resonance imaging, in which we could confirm the intact airway. Although the size of the ranula noted in our fetus was the smallest among the cases reported in the English literature, both of these imaging modalities clearly presented typical diagnostic features present on both ultrasound and magnetic resonance imaging. PMID- 20014016 TI - Risk of ischemic stroke in patients with Crohn's disease: a population-based nested case-control study. AB - BACKGROUND: Observational studies have linked Crohn's disease (CD) to an increased risk of venous thromboembolic complications. Case reports of ischemic stroke in CD patients have raised the question of a similar association, but data from observational studies are lacking. METHODS: Using data from the UK General Practice Research Database we conducted a nested case-control analysis within a population-based cohort of 8054 patients with and 161,078 patients without CD. A total of 1748 cases of ischemic stroke were identified to whom 17,348 controls were matched on age, sex, and year of cohort entry. Adjusted odds ratios (ORs) of ischemic stroke associated with CD were calculated by conditional logistic regression. Stratified analyses were performed for age and sex. RESULTS: While CD was not associated with an overall increased risk of ischemic stroke (OR 1.10, 95% confidence interval [CI] 0.85-1.43), stratified analyses revealed an increase in risk in younger patients (<50 years: OR 2.93; 95% CI 1.44-5.98) but not in elderly patients (> or =50 years: OR 0.99; 95% CI 0.75-1.30; P for interaction <0.01). The interaction with age remained statistically significant even after changing the cutoff value for the younger and older age group to 45, 55, or 60 years in a sensitivity analysis. There was no interaction with sex (P = 0.79). CONCLUSIONS: The study indicates that younger patients with CD may be under an increased risk of ischemic stroke. PMID- 20014017 TI - Low-dose naltrexone for treatment of duodenal Crohn's disease in a pediatric patient. PMID- 20014018 TI - Characterization of the novel ST2/IL-33 system in patients with inflammatory bowel disease. AB - BACKGROUND: ST2 has been proposed to be a regulator of inflammation and Th1/Th2 balance. ST2L is the IL-33 membrane receptor and belongs to the IL-1R family. The soluble variant, ST2s, is identical to the extracellular region of ST2L and competes for IL-33 binding, inhibiting receptor signaling. Although ST2s has been associated with inflammatory processes in patients with sepsis, trauma, asthma, and autoimmunity, until now there are no reported studies showing the role of ST2/IL-33 in inflammatory bowel disease (IBD). METHODS: Expression of ST2 and IL 33 was determined in serum and colonic biopsies from IBD patients. ST2 transcript and protein was determined by reverse-transcription polymerase chain reaction (RT PCR) and enzyme-linked immunosorbent assay (ELISA)/immunoblot, respectively, and IL-33 protein by ELISA. Intestinal mucosa localization of ST2 and IL-33 was conducted by immunofluorescence. RESULTS: ST2s transcript in the colonic mucosa was mainly expressed in UC patients rather than Crohn's disease or control; however, ST2L mRNA remained constant in all samples. Total ST2 protein was significantly higher in mucosa samples from patients with active UC, with a predominant induction of ST2s that strongly correlates with serum ST2 levels. Mucosa IL-33 levels were higher in UC patients and serum levels were barely detected in all patient groups. ST2 and IL-33 are both abundantly expressed in the cytoplasm of epithelial cells of control subjects; however, in ulcerative colitis patients ST2 decreases and IL-33 showed cytoplasm-nuclear redistribution. CONCLUSIONS: The novel association between the ST2/IL-33 system and IBD seems to identify that variations in this axis might regulate the inflammatory process in these diseases. PMID- 20014019 TI - Genome wide association (GWA) predictors of anti-TNFalpha therapeutic responsiveness in pediatric inflammatory bowel disease. AB - BACKGROUND: Interindividual variation in response to anti-TNFalpha therapy may be explained by genetic variability in disease pathogenesis or mechanism of action. Recent genome-wide association studies (GWAS) in inflammatory bowel disease (IBD) have increased our understanding of the genetic susceptibility to IBD. The aim was to test associations of known IBD susceptibility loci and novel "pharmacogenetic" GWAS identified loci with primary nonresponse to anti-TNFalpha in pediatric IBD patients and develop a predictive model of primary nonresponse. METHODS: Primary nonresponse was defined using the Harvey Bradshaw Index (HBI) for Crohn's disease (CD) and partial Mayo score for ulcerative colitis (UC). Genotyping was performed using the Illumina Infinium platform. Chi-square analysis tested associations of phenotype and genotype with primary nonresponse. Genetic associations were identified by testing known IBD susceptibility loci and by performing a GWAS for primary nonresponse. Stepwise multiple logistic regression was performed to build predictive models. RESULTS: Nonresponse occurred in 22 of 94 subjects. Six known susceptibility loci were associated with primary nonresponse (P < 0.05). Only the 21q22.2/BRWDI loci remained significant in the predictive model. The most predictive model included 3 novel "pharmacogenetic" GWAS loci, the previously identified BRWD1, pANCA, and a UC diagnosis (R(2) = 0.82 and area under the curve [AUC] = 0.98%). The relative risk of nonresponse increased 15-fold when number of risk factors increased from 0-2 to > or =3. CONCLUSIONS: The combination of phenotype and genotype is most predictive of primary nonresponse to anti-TNFalpha in pediatric IBD. Defining predictors of response to anti-TNFalpha may allow the identification of patients who will not benefit from this class of therapy. PMID- 20014020 TI - Elevated IL-13Ralpha2 in intestinal epithelial cells from ulcerative colitis or colorectal cancer initiates MAPK pathway. AB - BACKGROUND: Chronic inflammation in ulcerative colitis (UC) is a sizeable risk factor for colorectal cancer (CRC). Interleukin-13 (IL-13) is elevated in the UC mucosa and may induce dysregulated signaling in neighboring intestinal epithelial cells (IECs) and thus function as a tumorogenic cytokine. METHODS: Expression of IL-13 receptor chains on IECs obtained from control or chronically inflamed mucosa and colonic tumors was quantified by flow cytometry and immunoblot. IL 13Ralpha1 and IL-13Ralpha2 expression was significantly increased on IEC from UC and CRC patients compared to control and Crohn's disease (CD) subjects. Purified IEC from these subjects and cell lines expressing varying ratios of IL-13Ralpha1 and IL-13Ralpha2 chains were stimulated with IL-13 in vitro to investigate by immunoblot the activation of the signal transducer and activator of transcription 6 (STAT6) and mitogen activated protein kinase (MAPK) signaling pathways. RESULTS: Despite similarly elevated receptor expression in UC and CRC, IL-13 does not activate the STAT6 or MAPK pathways in UC, while in colonic tumors only the STAT6 pathway is activated. Using neutralizing antibodies and cell lines expressing a range of surface densities for IL-13Ralpha1 and IL-3Ralpha2, we demonstrate that IL-13Ralpha2 serves a dual role, initiating MAPK signaling at low concentrations and as an inhibitory, decoy receptor at high IL-13Ralpha2 to IL-13Ralpha1 ratios. CONCLUSIONS: IL-13Ralpha2 is both a decoy receptor and induces MAPK signal transduction, depending on its relative expression and the local concentration of IL-13, which together modulate the balance and intensity of the signaling pathways initiated in UC and CRC. PMID- 20014021 TI - Efficacy and safety of certolizumab pegol induction therapy in an unselected Crohn's disease population: results of the FACTS survey. AB - BACKGROUND: Switzerland was the first country to approve certolizumab pegol (Cimzia, CZP) for the treatment of patients with moderate to severe Crohn's disease (CD) in September 2007. This phase IV study aimed to evaluate the efficacy and safety of CZP in a Swiss multicenter cohort of practice-based patients. METHODS: Baseline and Week 6 evaluation questionnaires were sent to all Swiss gastroenterologists in hospitals and private practices. Disease activity was assessed with the Harvey-Bradshaw Index (HBI) and adverse events were evaluated according to WHO guidelines. RESULTS: Fifty patients (31 women, 19 men) were included; 56% had complicated disease (stricture or fistula) and 52% had undergone prior CD-related surgery. All patients had prior exposure to systemic steroids, 96% to immunomodulators, 78% to infliximab, and 50% to adalimumab. A significant decrease in HBI was observed at Week 6 (versus Week 0) following induction therapy with CZP 400 mg subcutaneously at Weeks 0, 2, and 4 (12.6 +/- 4.7 Week 0 versus 6.2 +/- 4.4 Week 6, P < 0.001). Response and remission rates at Week 6 were 54% and 40%, respectively. We identified 8/11 CD patients undergoing a 50% fistula response (P = 0.021). The frequency of adverse drug reactions attributed to CZP was 6%. CZP was continued in 80% of patients beyond Week 6. CONCLUSIONS: In a population of CD patients with complicated disease behavior, CZP induced a response and remission in 54% and 40% of patients, respectively. This series provides the first evidence of the effectiveness of CZP in perianal fistulizing CD. PMID- 20014022 TI - Localization of the lipopolysaccharide recognition complex in the human healthy and inflamed premature and adult gut. AB - BACKGROUND: Microbiota in the intestinal lumen provide an abundant source of potentially detrimental antigens, including lipopolysaccharide (LPS), a potent immunostimulatory product of Gram-negative bacteria recognized by the host via TLR-4 and MD-2. An aberrant immune response to LPS or other bacterial antigens has been linked to inflammatory bowel disease (IBD) and necrotizing enterocolitis (NEC). METHODS: We investigated which cells express MD-2 in the normal and inflamed ileum from neonates and adults by immunohistochemistry. Moreover, MD-2 and TLR4 mRNA expression in normal adult ileum was studied by reverse transcription polymerase chain reaction (RT-PCR) on cells isolated by laser capture microdissection. RESULTS: Premature infants did not show MD-2 expression either in epithelial cells or in the lamina propria. Similarly, MD-2 was absent in epithelial cells and lamina propria inflammatory cells in preterm infants with NEC. MD-2 protein in the healthy term neonatal and adult ileum was predominantly expressed by Paneth cells and some resident inflammatory cells in the lamina propria. MD-2 and TLR-4 mRNA expression was restricted to crypt cells. Also in IBD, Paneth cells were still the sole MD-2-expressing epithelial cells, whereas inflammatory cells (mainly plasma cells) were responsible for the vast majority of the MD-2 expression. CONCLUSIONS: The absence of MD-2 in the immature neonatal gut suggests impaired LPS sensing, which could predispose neonates to NEC upon microbial colonization of the immature intestine. The apparent expression of MD-2 by Paneth cells supports the critical concept that these cells respond to luminal bacterial products in order to maintain homeostasis with the intestinal microbiota in vivo. PMID- 20014023 TI - In memoriam: Charles Tanford(1921-2009). PMID- 20014024 TI - Helix propensities calculations for amino acids in alanine based peptides using Jarzynski's equality. AB - Jarzynski's equality (Jarzynski, Phys Rev E 1997; 56:5018 and Jarzynski, Phys Rev Lett 1997; 78:2690) relates equilibrium free energy differences between two states A and B to the work done when the system is driven repeatedly and irreversibly from an equilibrium state A to equilibrium state B. We present calculations of helix propensities using a novel procedure based on this equality. In particular, a work probability distribution is built based on combinations of multi-step trajectories that give representative work distributions without requiring computing an unreasonable large number of trajectories between states. A small number of trajectories (15) are used to construct a distribution that contains millions of work values. This distribution is used to calculate DeltaG(AB) using Jarsynski's equation. To apply and test this method, we used as a model system a dodeca-alanine helix, analyzing its extension using mechanical force. This helix was used as the basis of a host guest system in which two of the 12 residues are substituted by some other amino acid (as the guests). The differences between the unfolding free energies of the substituted peptides and the all-alanine peptide provided values for DeltaDeltaG that can be interpreted as the helix propensities of each amino acid. Results show good correlation with the experimental measurements of Baldwin and coworkers (Chakrabartty et al., Protein Sci 1994; 3:843-852). PMID- 20014025 TI - Protein structure prediction begins well but ends badly. AB - The accurate prediction of protein structure, both secondary and tertiary, is an ongoing problem. Over the years, many approaches have been implemented and assessed. Most prediction algorithms start with the entire amino acid sequence and treat all residues in an identical fashion independent of sequence position. Here, we analyze blind prediction data to investigate whether predictive capability varies along the chain. Free modeling results from recent critical assessment of techniques for protein structure prediction (CASP) experiments are evaluated; as is the most up-to-date data from EVA, a fully automated blind test of secondary structure prediction servers. The results demonstrate that structure prediction accuracy is dependent on sequence position. Both secondary structure and tertiary structure predictions are more accurate in regions near the amino(N) terminus when compared with analogous regions near the carboxy(C)-terminus. Eight of 10 secondary structure prediction algorithms assessed by EVA perform significantly better in regions at the N-terminus. CASP data shows a similar bias, with N-terminal fragments being predicted more accurately than fragments from the C-terminus. Two analogous fragments are taken from each model, the N terminal fragment begins at the start of the most N-terminal secondary structure element (SSE), whereas the C-terminal fragment finishes at the end of the most C terminal SSE. Each fragment is locally superimposed onto its respective native fragment. The relative terminal prediction accuracy (RMSD) is calculated on an intramodel basis. At a fragment length of 20 residues, the N-terminal fragment is predicted with greater accuracy in 79% of cases. PMID- 20014026 TI - Formulation development of antibodies using robotic system and high-throughput laboratory (HTL). AB - Since each antibody has its unique physical chemical properties, optimal formulation for one antibody is likely not applicable for the others. To rapidly screen multiple antibody formulations, an automated system was constructed to perform sample preparation, testing, and data management. Using the automatic system, up to 500 liquid formulations can be prepared in deep well microplates and further distributed into standard microplates that can be stored under different stress conditions for degradation studies. In addition, the system can also be used to prepare samples in microplates for different analytical measurements such as UV spectroscopy, turbidity, dynamic light scattering (DLS), SEC-HPLC, RP-HPLC and CEX-HPLC, and automated lab-on-a-chip platform (ALP). The data generated using different techniques in the automatic system were comparable to those of the classical approaches. PMID- 20014027 TI - Solution structure and dynamics of human ubiquitin conjugating enzyme Ube2g2. AB - Ube2g2 is an E2 enzyme which functions as part of the endoplasmic reticulum associated degradation (ERAD) pathway responsible for identification and degradation of misfolded proteins in the endoplasmic reticulum. In tandem with a cognate E3 ligase, Ube2g2 assembles K48-linked polyubiquitin chains and then transfers them to substrate, leading ultimately to proteasomal degradation of the polyubiquitin-tagged substrate. We report here the solution structure and backbone dynamics of Ube2g2 solved by nuclear magnetic resonance spectroscopy. Although the solution structure agrees well with crystallographic structures for the E2 core, catalytically important loops (encompassing residues 95-107 and 130 135) flanking the active site cysteine are poorly defined. (15)N spin relaxation and residual dipolar coupling analysis directly demonstrates that these two loops are highly dynamic in solution. These results suggest that Ube2g2 requires one or more of its protein partners, such as cognate E3, acceptor ubiquitin substrate or thiolester-linked donor ubiquitin, to assume its catalytically relevant conformation. Within the NMR structural ensemble, interactions were observed between His94 and the highly mobile loop residues Asp98 and Asp99, supporting a possible role for His94 as a general base activated by the carboxylate side chains of Asp98 or Asp99. PMID- 20014028 TI - The influence of captive adolescent male chimpanzees on wounding: management and welfare implications. AB - Adolescence, the period lasting from the onset of puberty to the emergence of physical and sexual maturity, is a period of social change for many species including chimpanzees. Several reports have implicitly linked the physiological changes that occur during male chimpanzee adolescence to significant disruption in the social group, which in turn may result in serious agonism and wounding. To assess the association between adolescent males and wounding rates, 38 institutions housing 399 chimpanzees among 59 social groups, recorded all wounds incurred by chimpanzees over a 6-month period. The rate of wounding did not differ between groups with or without adolescent males. Adolescent males received the most wounds, but were no more likely to cause wounds than group members of any other sex-age class. Social groups with multiple adult males experienced lower wounding rates than those with a single adult male. Results indicate that (1) adolescent male chimpanzees may receive, but not inflict, more wounds than chimpanzees in other sex-age classes; and (2) management strategies that support natural social groupings may control and limit group agonism. PMID- 20014029 TI - Osmolytes modulate conformational exchange in solvent-exposed regions of membrane proteins. AB - Site-directed spin labeling (SDSL) was used to investigate local structure and conformational exchange in two bacterial outer-membrane TonB-dependent transporters, BtuB and FecA. Protecting osmolytes, such as polyethylene glycols (PEGs) are known to modulate a substrate-dependent conformational equilibrium in the energy coupling motif (Ton box) of BtuB. Here, we demonstrate that a segment that is N-terminal to the Ton box in BtuB, is in conformational exchange between ordered and disordered states with or without substrate. Protecting osmolytes shift this equilibrium to favor the more ordered, folded state. However, a segment of BtuB that is C-terminal to the Ton box that is not solvent exposed is insensitive to PEGs. Protecting osmolytes also modulate a conformational equilibrium in the Ton box of FecA, with larger molecular weight PEGs producing the largest shifts in the conformational free energy. These data indicate that solvent-exposed regions of these transporters undergo conformational exchange and that regions of these transporters that are involved in protein-protein interactions sample multiple conformational substates. The sensitivity to solute provides an explanation for differences seen between two high-resolution structures of BtuB, which each likely represent one conformation from a subset of states that are normally sampled by the protein. This work also illustrates how SDSL and osmolytes may be used to characterize and quantitate conformational equilibria in membrane proteins. PMID- 20014030 TI - Versatile modes of peptide recognition by the ClpX N domain mediate alternative adaptor-binding specificities in different bacterial species. AB - ClpXP, an AAA+ protease, plays key roles in protein-quality control and many regulatory processes in bacteria. The N-terminal domain of the ClpX component of ClpXP is involved in recognition of many protein substrates, either directly or by binding the SspB adaptor protein, which delivers specific classes of substrates for degradation. Despite very limited sequence homology between the E. coli and C. crescentus SspB orthologs, each of these adaptors can deliver substrates to the ClpXP enzyme from the other bacterial species. We show that the ClpX N domain recognizes different sequence determinants in the ClpX-binding (XB) peptides of C. crescentus SspBalpha and E. coli SspB. The C. crescentus XB determinants span 10 residues and involve interactions with multiple side chains, whereas the E. coli XB determinants span half as many residues with only a few important side chain contacts. These results demonstrate that the N domain of ClpX functions as a highly versatile platform for peptide recognition, allowing the emergence during evolution of alternative adaptor-binding specificities. Our results also reveal highly conserved residues in the XB peptides of both E. coli SspB and C. crescentus SspBalpha that play no detectable role in ClpX-binding or substrate delivery. PMID- 20014032 TI - Getting it right. PMID- 20014031 TI - Adding a Nursing Information Technology Subscale to the Practice Environment Scale of the Nursing Work Index. AB - In the past decade, the use of information technology (IT) to support healthcare delivery has greatly expanded. Introducing new clinical information systems and updating existing systems require continued learning and training among registered nurses (RNs) and other clinicians. Consequently, a 5-item measure was designed as a subscale to the Practice Environment Scale of the Nursing Work Index. This new subscale, the Nursing Information Technology Subscale (NITS) measures RNs' perception of the extent to which IT in their practice environment supports patient care delivery. A psychometric evaluation of the NITS was conducted at 8 Veterans Affairs hospitals. The findings demonstrated that this brief subscale has acceptable reliability as well as convergent and discriminant validity. PMID- 20014034 TI - Dissymmetries in fluorescence excitation and emission from single chiral molecules. AB - Chirality in molecular systems plays profoundly important roles in chemistry and physics. Most chemistry students are introduced to the concept of chirality through demonstrations of the interaction of chiral molecules with polarized light manifested as an "optical rotation" leading to the "(+)" and "(-)" [or dextrorotatory (d-) and levorotatory (l-)] designations of chiral compounds, with the subsequent determination of absolute stereochemical configuration by chemical or physical means enabling application of the familiar "R" and "S" labels. Although the intrinsic molecular parameters that control the dissymmetric light matter interaction in chiral systems are well understood, we have only recently begun to ask questions regarding the role of local molecular environment and hidden heterogeneities associated with the ensemble-averaged molecular chiroptical response. In this mini-review, we discuss some of our recent research on application of single-molecule spectroscopy as a tool for probing heterogeneities and fluctuations of chiroptical dissymmetries in condensed phase. PMID- 20014035 TI - Correlation between R/S enantiomer ratio of lansoprazole and CYP2C19 activity after single oral and enteral administration. AB - The purpose of this study was to investigate whether CYP2C19 activity can be estimated from plasma concentrations of lansoprazole enantiomers 4 h (C(4h)) after single administration by oral and enteral routes. Sixty-nine subjects, 22 homozygous extensive metabolizers (homEMs), 32 heterozygous EMs (hetEMs), and 15 poor metabolizers (PMs), participated in the study. After a single oral or enteral dose of racemic lansoprazole (30 mg), plasma concentrations of lansoprazole enantiomers were measured 4 h postdose. The R/S ratio of lansoprazole at 4 h differed significantly among the three groups (P < 0.0001) regardless of the administration route. The R/S ratio of lansoprazole in CYP2C19 PMs ranged from 3.0 to 13.7, whereas in homEMs and hetEMs the ratio ranged from 8.6 to 90 and 2.1 to 122, respectively. The relationship between (S)-lansoprazole concentration and R/S ratio of lansoprazole at C(4h) is given by the following formula: log(10) [R/S ratio] = 2.2 - 0.64 x log(10) [C(4h) of (S)-lansoprazole] (r = 0.867, P < 0.0001). Thus, phenotyping CYP2C19 using the R/S enantiomer ratio of lansoprazole seems unlikely. However, to obtain a pharmacological effect similar to that in CYP2C19 PMs, we can presume that lansoprazole has a sufficient effect in the patient with an R/S enantiomer ratio at 4 h < or = 13.70 and (S) lansoprazole concentration at 4 h > or = 50 ng/ml. PMID- 20014036 TI - Axially chiral N-(o-aryl)-4-hydroxy-2-oxazolidinone derivatives from diastereoselective reduction of N-(o-aryl)-2,4-oxazolidinediones: thermally interconvertible atropisomers via ring-chain-ring tautomerization. AB - The reduction of the axially chiral N-(o-aryl)-5,5-dimethyl-2,4-oxazolidinediones by NaBH(4) yielded axially chiral N-(o-aryl)-4-hydroxy-5,5-dimethyl-2 oxazolidinone enantiomers having a chiral center at C-4, with 100% diastereoselectivity as has been shown by their (1)H and (13)C NMR spectra and by enantioselective HPLC analysis. The resolved enantiomeric isomers were found to interconvert thermally through an aldehyde intermediate formed upon ring cleavage via a latent ring-chain-ring tautomerization. It was found that the rate of enantiomerization depended on the size and the electronic effect of the ortho substituent present on the aryl ring bonded to the nitrogen of the heterocycle. PMID- 20014037 TI - Borane-mediated asymmetric reduction of acetophenone by enantiopure aminonaphthols and aminoalcohols as catalytic source. AB - Practical, cheap, and stereoselective synthetic methods were applied to the preparation of novel 1-(aminoalkyl)naphthol and gamma-aminoalcohol tridentate ligands. The ligands obtained were conveniently applied with good results as catalytic sources in the borane-mediated enantioselective reduction of acetophenone with borane dimethylsulfide. Conformational analysis through molecular modeling allows the rationalization of observed stereochemical outcomes. PMID- 20014038 TI - Modification of chiral dimethyl tartrate through transesterification: immobilization on POSS and enantioselectivity reversal in sharpless asymmetric epoxidation. AB - Modification of dimethyl tartrate has been investigated through transesterification with aminoalcohols to provide reactive functionalities for the covalent bonding of chiral tartrate to polyhedral oligomeric silsesquioxanes. The transesterification of dimethyl tartrate has been widely studied using different catalytic systems and reaction conditions. Through the proper selection of both the catalytic system and the reaction conditions, it is possible to achieve monosubstituted or bis-substituted tartrate derivatives as sole products. All the intermediate chiral tartrate-derived ligands were successfully used in the homogeneous enantioselective epoxidation of allylic alcohols providing moderate enantiomeric excess over the products. Attached amine groups have been used to support the modified tartrate ligands on to a haloaryl-functionalized silsesquioxane moiety. This final chiral tartrate ligand displays reverse enantioselectivity in the asymmetric epoxidation of allylic alcohols with regard to the starting dimethyl tartrate ligand, both molecules having the same chiral sign. However, the POSS-containing ligand can be easily recovered in almost quantitative yield and reused in asymmetric epoxidation reactions. In addition, recovered silsesquioxane-pendant ligand, though displaying decreasing catalytic activity in recycling epoxidation tests, showed very stable enantioselective behavior. PMID- 20014039 TI - Cloning and characterization of a glycerol-3-phosphate dehydrogenase (NAD+) gene from the halotolerant yeast Pichia farinosa. AB - In this study, a novel glycerol-3-phosphate dehydrogenase (NAD(+)) (EC1.1.1.8) gene (PfGPD) was cloned from halotolerant yeast Pichia farinosa, using degenerate reverse transcription (RT)-PCR and rapid amplification of cDNA ends (RACE) methods. The full-length cDNA of the PfGPD gene is 1403 bp, which has an open reading frame (ORF) encoding 370 amino acids. PfGPD is conserved with other glycerol-3-phosphate dehydrogenase genes and presents a single copy in the P. farinosa genome. Transcriptional analysis revealed that PfGPD gene expression level was high after 2 h induction in a hyperosmotic environment containing 2 M NaCl and returned to normal within 6 h. These results suggest that the PfGPD gene is induced by salt stress. Yeast complementation experiment indicated that PfGPD complements gpd1 mutation in S. cerevisiae. The Accession No. for PfGPD in GenBank is EF601986. PMID- 20014040 TI - Successful clinical implementation of corneal epithelial stem cell therapy for treatment of unilateral limbal stem cell deficiency. AB - The corneal epithelium is maintained by a population of stem cells known as limbal stem cells (LSCs) due to their location in the basal layer of the outer border of the cornea known as the limbus. Treatment of limbal stem cell deficiency (LSCD) has been achieved with transplantation of ex vivo expanded LSCs taken from a small biopsy of limbus. This is a relatively new technique, and as such, specific national or international guidance has yet to be established. Because of the lack of such specific guidance, our group has sought to minimize any risk to the patient by adopting certain modifications to the research methodologies in use at present. These include the replacement of all non-human animal products from the culture system and the production of all reagents and cultures under Good Manufacturing Practice conditions. In addition, for the first time, a strictly defined uniform group of patients with total unilateral LSCD and no other significant ocular conditions has been used to allow the success or failure of treating LSCD to be attributable directly to the proposed stem cell therapy. A prospectively designed study with strict inclusion and exclusion criteria was used to enroll patients from our database of patients with unilateral LSCD. Eight eyes of eight consecutive patients with unilateral total LSCD treated with ex vivo expanded autologous LSC transplant on human amniotic membrane (HAM) with a mean follow-up of 19 (RANGE) months were included in the study. Postoperatively, satisfactory ocular surface reconstruction with a stable corneal epithelium was obtained in all eyes (100%). At last examination, best corrected visual acuity improved in five eyes and remained unchanged in three eyes. Vision impairment and pain scores improved in all patients (p < .05). This study demonstrates that transplantation of autologous limbal epithelial stem cells cultured on HAM without the use of non-human animal cells or products is a safe and effective method of reconstructing the corneal surface and restoring useful vision in patients with unilateral total LSCD. PMID- 20014041 TI - Identification and characterization of the type 2C protein phosphatase Ptc4p in the human fungal pathogen Candida albicans. AB - Type 2C protein phosphatases (PP2C) are monomeric enzymes and their activities require the presence of magnesium or manganese ions. There are seven PP2C genes, named from PTC1 to PTC7, in Saccharomyces cerevisiae. In the current study we identified the CaPTC4 gene in Candida albicans and demonstrated that the CaPtc4p protein is a typical PP2C enzyme, which is highly conserved in fungal species. Deletion of CaPTC4 renders Candida cells sensitive to sodium and potassium ions as well as to antifungal azole drugs. In addition, we have shown that CaPtc4p is localized in the mitochondrion, suggesting that CaPtc4p is likely to be involved in the regulation of a mitochondrial function related to ion homeostasis. PMID- 20014042 TI - Prolonged absorption of antimony(V) by the oral route from non-inclusion meglumine antimoniate-beta-cyclodextrin conjugates. AB - The orally active composition comprising meglumine antimoniate (MA) and beta cyclodextrin (beta-CD) differs markedly from conventional drug-CD complexes, since it combines a water-soluble drug and a hydrophilic CD. In order to obtain insights into the mechanism(s) responsible for the improved oral delivery of the drug, physicochemical and pharmacokinetic studies were carried out. The composition investigated here was prepared at a 7:1 antimony(Sb)/beta-CD molar ratio, a condition that improves its solubility in water and allows the oral administration of a high dose of Sb in large animals. It was characterized by circular dichroism, (1)H-NMR, ESI-MS and photon correlation spectroscopy. Pharmacokinetic data were obtained in Beagle dogs after oral administration of the composition at 100 mg Sb/kg. (1)H-NMR and ESI-MS data supported the formation of non-inclusion complexes between MA and beta-CD. Sub-micron assemblies were also evidenced that slowly dissociate and presumably release the MA drug, upon reconstitution of the composition in water. Pharmacokinetic studies of MA and MA/beta-CD in dogs showed a prolongation of the serum mean residence time of Sb from 4.1 to 6.8 h, upon complexation of MA with beta-CD. Evidence was also obtained that Sb remains essentially under the form of pentavalent Sb-meglumine complex, following gastro-intestinal absorption from the MA/beta-CD composition. In conclusion, the present data support the model that the sustained drug release property of 7:1 MA/beta-CD composition resulted in the prolongation of MA absorption by the oral route and, consequently, in the increase of the drug mean residence time in serum. PMID- 20014043 TI - Amino acid residues involved in ligand preference of the Snf3 transporter-like sensor in Saccharomyces cerevisiae. AB - Snf3 is a plasma membrane protein in Saccharomyces cerevisiae able to sense the presence of glucose. Although the Snf3 protein does not transport sugars, it shares sequence similarity with various glucose transporters from other organisms. We investigated the sugar specificity/preferences of Snf3. The ability of cells to sense sugars in vivo was monitored by following the degradation of the Mth1 protein, an early event in the signal pathway. Our study reveals that Snf3, in addition to glucose, also senses fructose and mannose, as well as the glucose analogues 2-deoxyglucose, 3-O-methylglucoside and 6-deoxyglucose. The signalling proficiency of a non-phosphorylatable analogue strongly supports the notion that sensing through Snf3 does not require sugar phosphorylation. Sequence comparisons of Snf3 to glucose transporters indicated amino acid residues possibly involved in sensing of sugars other than glucose. By site-specific mutagenesis of the structural gene, roles of specific residues in Snf3 could be established. Change of isoleucine-374 to valine in transmembrane segment 7 of Snf3 partially abolished sensing of fructose and mannose, while mutagenesis causing a change of phenylalanine-462 to tyrosine in transmembrane segment 10 of Snf3 abolished sensing of fructose. Neither of these amino acid changes affected the ability of Snf3 to sense glucose, nor did they permit Snf3 to sense galactose. These data indicate a similarity between a ligand binding site of the sensor Snf3 and binding sites used for facilitated hexose transport in the GLUT proteins. PMID- 20014044 TI - Most, but not all, yeast strains in the deletion library contain the [PIN(+)] prion. AB - The yeast deletion library is a collection of over 5100 single gene deletions that has been widely used by the yeast community. The presence of a non-Mendelian element, such as a prion, within this library could affect the outcome of many large-scale genomic studies. We previously showed that the deletion library parent strain contained the [PIN(+)] prion. [PIN(+)] is the misfolded infectious prion form of the Rnq1 protein that displays distinct fluorescent foci in the presence of RNQ1-GFP and exists in different physical conformations, called variants. Here, we show that over 97% of the library deletion strains are [PIN(+)]. Of the 141 remaining strains that have completely (58) or partially (83) lost [PIN(+)], 139 deletions were able to efficiently maintain three different [PIN(+)] variants despite extensive growth and storage at 4 degrees C. One strain, cue2Delta, displayed an alteration in the RNQ1-GFP fluorescent shape, but the Rnq1p prion aggregate shows no biochemical differences from the wild type. Only strains containing a deletion of either HSP104 or RNQ1 are unable to maintain [PIN(+)], indicating that 5153 non-essential genes are not required for [PIN(+)] propagation. PMID- 20014046 TI - Infrared multiple photon dissociation spectroscopy of sodium and potassium chlorate anions. AB - The structures of gas-phase, metal chlorate anions with the formula [M(ClO(3))(2)](-), M = Na and K, were determined using tandem mass spectrometry and infrared multiple photon dissociation (IRMPD) spectroscopy. Structural assignments for both anions are based on comparisons of the experimental vibrational spectra for the two species with those predicted by density functional theory (DFT) and involve conformations that feature either bidentate or tridentate coordination of the cation by chlorate. Our results strongly suggest that a structure in which both chlorate anions are bidentate ligands is preferred for [Na(ClO(3))(2)](-). However, for [K(ClO(3))(2)](-) the best agreement between experimental and theoretical spectra is obtained from a composite of predicted spectra for which the chlorate anions are either both bidentate or both tridentate ligands. In general, we find that the overall accuracy of DFT calculations for prediction of IR spectra is dependent on both functional and basis set, with best agreement achieved using frequencies generated at the B3LYP/6-311+g(3df) level of theory. PMID- 20014045 TI - A novel kanamycin/G418 resistance marker for direct selection of transformants in Escherichia coli and different yeast species. AB - We have developed a set of cloning vectors possessing a modified Tn903 kanamycin resistance gene that enables the selection of both kanamycin-resistant transformants in Escherichia coli and G418-resistant transformants in the yeasts Saccharomyces cerevisiae, Hansenula polymorpha and Pichia pastoris. Expression of this gene in yeast is controlled by the H. polymorpha glyceraldehyde-3-phosphate dehydrogenase promoter, while expression in E. coli is governed by an upstream E. coli lacZ promoter. Applicability of the vectors for gene disruption in H. polymorpha and S. cerevisiae was demonstrated by inactivation of the HpMAL1 and URA3 genes, respectively. One of the vectors possesses a H. polymorpha ARS allowing plasmid maintenance in an episomal state. The small size of the vectors (2-2.5 kb) makes them convenient for routine DNA cloning. In addition, we report a novel approach for construction of gene disruption cassettes. PMID- 20014047 TI - Agents of support: psychometric properties of the Cancer Perceived Agents of Social Support (CPASS) questionnaire. AB - OBJECTIVE: The current study presents the development and the evaluation of the psychometric properties of the Cancer Perceived Agents of Social Support (CPASS). The CPASS is a new self-rating instrument devised in order to enable both cancer patients and their spouses to report on the level of perceived social support they get. The CPASS evaluates the support given by different agents of support (spouse, family, friends and spiritual or religious beliefs) in several dimensions (emotional, cognitive and instrumental). METHODS: The study sample comprised 662 cancer patients and their spouses recruited during a routine medical evaluation from three major cancer centers in Israel. The participants completed the CPASS and two other standardized instruments: The ENRICH Marital Satisfaction Scale (EMS) and the Brief Symptom Inventory (BSI). RESULTS: Principal component analysis confirmed a three-factor structure based upon the agent of support (spouse; friends/family; spiritual/religious beliefs). Cronbach's alpha coefficients for the agent of support indexes were high (0.80 0.95), while Cronbach's alpha levels for the kind of support were lower (0.45 0.72). Smallest Space Analysis (SSA) also confirmed the theoretical structure of the CPASS. Pearson correlation coefficients to the other study variables were high and significant. CONCLUSIONS: As a whole, the CPASS was found to be a valid tool for the current Israeli sample. Theoretical and practical conclusions and socio-cultural implications are discussed. PMID- 20014049 TI - Study of onset time-shift and injection duration in DCE-MRI: a comparison of a reference region model with the general kinetic model. AB - In dynamic contrast-enhanced MR imaging (DCE-MRI), sampling of the arterial input function (AIF) is required for analysis using the general kinetic model (GKM). Alternatively, the recently proposed reference region model (RRM) may be employed to avoid the need of acquiring the AIF. This study aimed to evaluate the influence of the AIF onset-time shift and the injection duration, under various sampling intervals, on physiological parameter estimation in DCE-MRI using the GKM, and to compare the performance between GKM and RRM. Computer simulations were performed to assess the mean error (ME) and coefficient of variation (CoV) of K(trans,TOI) and v(e,TOI) from shifted and dispersed AIF with temporal resolution of 1, 5 and 10 s. With 5-s sampling, the maximal ME of K(trans,TOI) was roughly 22% for the GKM and 0.5% for the RRM. With 10-s sampling, they increased to around 28% and 0.7%, respectively. The maximal MEs of v(e,TOI) for all cases were under 5%. However, owing to the lower SNR in the reference region, the CoV obtained by the RRM were all higher than those by the GKM. The results suggested that with compromised temporal resolution, the RRM was relatively less sensitive to the AIF onset-time shift and the injection duration compared with the GKM. PMID- 20014050 TI - Use of psychotropic drugs and analgesics among users of antiobesity drugs--a population based study. AB - AIM: The aim of the present study was to investigate to what extent patients using prescription antiobesity drugs (orlistat, sibutramine and rimonabant) concomitantly or concurrently used psychotropic drugs and analgesics and the association between this drug use and the patients' gender and age. An additional aim was to investigate the sequence of drug therapy among users of both antiobesity drugs and antidepressants or antipsychotics, respectively. METHODS: Data were retrieved from the Norwegian Prescription Database (NorPD). All patients who had an antiobesity drug (ATC code A08A) dispensed from a Norwegian pharmacy between January 2004 and December 2007 were included in the study. RESULTS: One in four patients using antiobesity drugs had at least on one occasion used a psychotropic drug concomitantly. The most commonly used psychotropic drugs were anxiolytics/hypnotics/sedatives (17.7%) and antidepressants (14.7%). Analgesics were used by 36.2%. A significantly higher percentage of women used anxiolytics/hypnotics/sedatives (18.8% vs. 14.0%, p < 0.0005), antidepressants (16.1% vs. 9.5%, p < 0.0005), antipsychotics (4.0% vs. 2.9%, p < 0.0005) and analgesics (37.8% vs. 30.5%, p < 0.0005) concomitantly with antiobesity drugs when compared to men. One out of ten patients using sibutramine had at least on one occasion used an interacting drug concomitantly. CONCLUSION: Use of psychotropic drugs and analgesics among patients using antiobesity drugs is extensive, especially among women. Clinicians prescribing sibutramine should be more aware of drug interactions with other prescribed drugs. There is still insufficient information on psychiatric disorders among these patients. PMID- 20014051 TI - Topical bovine thrombin: a 21-year review of topical bovine thrombin spontaneous case safety reports submitted to FDA's Adverse Event Reporting System. AB - PURPOSE: To review topical bovine thrombin spontaneous adverse event (AE) reports that were forwarded to the US Food and Drug Administration's (FDA) Adverse Event Reporting System (AERS) between January 1986 and December 2006. METHODS: Forty one spontaneous AE reports were summarized for reported AE profile and chronological reporting patterns. Each AE report was adjudicated by a hematologist for the topical bovine thrombin product that was given and the AE(s) that were reported. AEs were grouped as allergic, coagulopathy/bleeding, and all other AEs combined. Grouped AE serial analyses were carried out using successive 3-year time increments between 1986 (the year an AE report was first noted for a bovine thrombin product) and 2006 (the first full year that was available at the time of initiation of the data summary). MAIN OUTCOME MEASURES: The primary outcome measures were every 3-year trend lines for all-AE reports, all reporters, and topical bovine thrombin brand mentions for 2 AE groups of interest (allergic events and coagulopathy/bleeding events). RESULTS: The all-AE spontaneous reporter trend showed a downward appearance for AE reporting activity that started in 1995-1998 and continued through 2004-2006. The all-AE reports trend showed two potential safety signals that could be identified serially: (1) a prominent 1989-1991 peak that was attributable to allergic events (in particular, anaphylaxis), and (2) a small 1995-2000 broad peak that was attributable in part to coagulopathy/bleeding events. Allergic events were predominantly reported with products approved prior to 1995, were not temporally associated with prior medical literature case reports, and continued to be forwarded to the FDA at low levels up to the end of this study in 2006. Coagulopathy/bleeding events were reported only with products approved prior to 1995, were temporally associated with medical literature case reports, and were not forwarded to the FDA after 2000. CONCLUSIONS: Overall, spontaneous AE reporting for topical bovine thrombin occurs at very low levels, and appears to have been decreasing since 1995. The serial reporting patterns for topical bovine thrombin are best explained as a strong safety signal for allergic events with ongoing, low level reporting, and a weak safety signal for coagulopathy/bleeding events that ceased on or before 2000. Although this descriptive trend analysis cannot measure associations or causation, the coagulopathy/bleeding signal may have been prompted by multiple, antecedent published case reports. The subsequent diminishment of signal attributed to thrombin likewise may coincide with lack of such reporting in larger follow-up clinical trials or, alternatively, in the introduction and growing market share of thrombin brands of greater purity. Currently marketed topical bovine thrombin formulations are rarely volunteered as possible causes of adverse events. PMID- 20014052 TI - MEKC: a powerful tool for the determination of amino acids in a variety of biomatrices. AB - The continued publication from year to year of new MEKC formulations for the analysis of amino acids provides evidence that both CZE and MEKC still have a great power of attraction in this area. The present review intends to cover the literature on MEKC of amino acids from 2007 until present: it has been planned to follow the same format of our previous review (Electrophoresis 2008, 29, 224-236) representing its ideal continuation. In addition to methodological developments, the more recent practical applications of MEKC procedures for the determination of amino acids in different matrices will also be described here as an evidence, once again, of the suitability of this technique on samples of different origin. PMID- 20014053 TI - Recent innovations in protein separation on microchips by electrophoretic methods: an update. AB - Lab-on-a-chip electrophoresis is becoming increasingly useful for protein analysis, thanks to recent developments in this field. This review is an update of the review we published at the start of 2008 [Peng, Y., Pallandre, A., Tran, N. T., Taverna, M., Electrophoresis 2008, 29, 156-177]. The superiority of polymers for the manufacture of analytical microchips has been confirmed. This trend implies several modifications to the processes previously used with glass/silicon chips and requires a better understanding of the interfacial phenomena of these materials. Significant progress in chip-based techniques for protein analysis has been made in the last 2 years. In addition to advances in traditional electrokinetic modes, counter-flow gradient focusing techniques have emerged as useful methods not only for separation, but also for the online preconcentration of samples. This review, with more than 175 references, presents recent advances and novel strategies for EOF measurement, surface treatment, sample pretreatment, detection and innovations relating to the different modes of separation. PMID- 20014054 TI - Recent developments in coupled SPE-CE. AB - This article presents an overview of coupled SPE-CE systems that have been reported in the literature between April 2007 and June 2009. The use of in-line and on-line SPE-CE is covered in this review. Special attention is paid to the use of monoliths and molecularly imprinted polymers in coupled SPE-CE systems. Application-oriented research is discussed in which in-line and on-line SPE-CE systems have been used in biomedical, pharmaceutical, environmental and food analysis. The SPE-CE studies are presented in tables, including information on sample type, SPE sorbent, detection and LOD. Finally, some future developments that may increase the applicability of coupled SPE-CE are highlighted. PMID- 20014055 TI - External validation of abbreviated versions of the activities-specific balance confidence scale in Parkinson's disease. AB - Balance confidence, measured using the Activity-Specific Balance Confidence Scale (ABC-16), can affect quality of life in individuals with Parkinson's disease (PD). Two abbreviated 6-item versions of the ABC-16 have been previously derived to measure balance confidence in an expedited fashion. This study aimed to validate these abbreviated versions and to determine their correlations with measures of balance and functional capacity. Eighty-nine individuals with PD participated in the study. Using the 25th percentile, five items were identified as the lowest scoring items (i.e., lowest balance confidence), and thus a 5-item version of the questionnaire was created. Internal consistency and the relationship with the ABC-16 were good for all abbreviated versions (Cronbach alpha > 0.93; ICC > 0.91). All versions correlated significantly with most measures of balance and function, with no differences between versions. An abbreviation of the ABC-16, using only five items, may prove useful in busy clinical and research settings. PMID- 20014056 TI - Sydenham's chorea in a girl with juvenile idiopathic arthritis treated with anti TNFalpha therapy. PMID- 20014058 TI - Enrichment and analysis of phosphopeptides under different experimental conditions using titanium dioxide affinity chromatography and mass spectrometry. AB - Titanium dioxide metal oxide affinity chromatography (TiO(2)-MOAC) is widely regarded as being more selective than immobilized metal-ion affinity chromatography (IMAC) for phosphopeptide enrichment. However, the widespread application of TiO(2)-MOAC to biological samples is hampered by conflicting reports as to which experimental conditions are optimal. We have evaluated the performance of TiO(2)-MOAC under a wide range of loading and elution conditions. Loading and stringent washing of peptides with strongly acidic solutions ensured highly selective enrichment for phosphopeptides, with minimal carryover of non phosphorylated peptides. Contrary to previous reports, the addition of glycolic acid to the loading solution was found to reduce specificity towards phosphopeptides. Base elution in ammonium hydroxide or ammonium phosphate provided optimal specificity and recovery of phosphorylated peptides. In contrast, elution with phosphoric acid gave incomplete recovery of phosphopeptides, whereas inclusion of 2,5-dihydroxybenzoic acid in the eluant introduced a bias against the recovery of multiply phosphorylated peptides. TiO(2)-MOAC was also found to be intolerant of many reagents commonly used as phosphatase inhibitors during protein purification. However, TiO(2)-MOAC showed higher specificity than immobilized gallium (Ga(3+)), immobilized iron (Fe(3+)), or zirconium dioxide (ZrO(2)) affinity chromatography for phosphopeptide enrichment. Matrix-assisted laser desorption/ionization mass spectrometry (MALDI MS) was more effective in detecting larger, multiply phosphorylated peptides than liquid chromatography/electrospray ionization tandem mass spectrometry (LC/ESI MS/MS), which was more efficient for smaller, singly phosphorylated peptides. PMID- 20014057 TI - Depression and major depressive disorder in patients with Parkinson's disease. AB - The prevalence of depression in Parkinson's disease (PD) varies greatly. In this study, we investigated major depressive disorder (MDD) and depressive symptoms without MDD in patients with PD. The psychopathological characteristics of depressive symptoms were assessed by a psychiatric interview. A total of 105 Japanese patients with PD without dementia were included. The Japanese version of the Beck Depression Inventory-II (BDI-II) with a cutoff score of 13/14 was used to screen for depression. Using a structured interview, a comprehensive psychiatric evaluation of patients with BDI-II scores >13 (high BDI patients) was completed using the criteria of the Diagnostic and Statistical Manual of Mental Disorders (DSM)-IV-TR. Forty patients (38%) had a BDI-II >13, but 29 did not show any depressed mood. Five cases met the criteria for MDD (three current, two past) and one patient was diagnosed with minor depressive disorder. A slight depressed mood that was associated with worrying about PD was seen in 6 of 34 patients without any depressive disorder and fluctuated with aggravation of PD symptoms in two of these patients. For the diagnosis of MDD, the number of positive items from the DSM-IV-TR definition of MDD is most important and useful for differentiating MDD and non-MDD. The low-prevalence rate of MDD in our patient population suggests that PD may be a psychological stressor for MDD, but does not necessarily induce MDD. PMID- 20014060 TI - Painful legs and moving toes following a traumatic medial plantar nerve injury. PMID- 20014061 TI - NMDA antagonist memantine improves levodopa-induced dyskinesias and "on-off" phenomena in Parkinson's disease. PMID- 20014062 TI - Photophobia in essential blepharospasm--a positron emission tomographic study. AB - To localize regional alterations in cerebral glucose metabolism in essential blepharospasm (EB) patients with photophobia. We have studied 22 EB patients by performing positron emission tomography and [(18)F]-fluorodeoxyglucose analysis. The patients were classified into two subgroups, namely, EB with photophobia (P group) and EB without photophobia (NP group), and compared with a healthy control group (n = 44). There were no significant differences between the two patient groups with respect to the severity of motor symptoms or the duration for which the condition persisted. The FDG-PET images were analyzed using the statistical parametric mapping software. As compared to the control group, the P group exhibited significant hypermetabolism in the thalamus (P = 0.002), while the NP group exhibited significant hypometabolism in the dorsal midbrain, especially, in the superior colliculus (P = 0.005). The P group exhibited significant hypermetabolism in the thalamus and the dorsal midbrain as compared to the NP group (P < 0.001). These findings suggest that photophobia in EB patients may be associated with abnormal hyperactivity in the thalamus. Either hyperactivity of the thalamus or hypoactivity of the superior colliculus, or both may be associated with excessive blinking in these patients. PMID- 20014063 TI - Mortality in Parkinson's disease. PMID- 20014064 TI - Myopathy causing camptocormia in idiopathic Parkinson's disease: a multidisciplinary approach. AB - Extreme forward flexion of the spine, named camptocormia (CC), and head drop syndrome (HD) may be among the most disabling symptoms in Parkinson's disease (PD). This study aims to eludicate the etiology of PD-associated CC and HD via a multidisciplinary approach (clinical examination, electromyography, MRI, genetic analysis, muscle morphology) centering on the histology of the paraspinal muscles. We studied 17 patients with the clinical diagnosis of PD and CC or head drop syndrome and six controls. We performed muscle biopsies of paraspinal muscles and deep neck extensor muscles. Mean age at onset of postural abnormality was 66 years and mean latency between onset of parkinsonian symptoms to first signs of CC or head drop was 7 years. The electromyogram of paraspinal muscles was abnormal in 13-14 patients. Histopathology revealed chronic myopathic changes in 14 of 17 biopsies, consisting of abnormal variation in fiber size, increase in internal nuclei, and increase in connective tissue, myofibrillar disarray and similarities to protein surplus myopathies. Interestingly, heterozygous variants in the Parkin gene were found in 2 of 9 investigated patients. We conclude that CC and HD in PD are predominantly myopathic. Aberrant protein aggregation may link PD and CC. PMID- 20014065 TI - Psychogenic movement disorder after a venlafaxine-induced dystonia. PMID- 20014067 TI - Pathogenesis of myeloma bone disease. AB - Multiple myeloma (MM) is the most common cancer to involve bone with up to 90% of patients developing bone lesions. The bone lesions are purely osteolytic in nature and do not heal in the vast majority of patients. Up to 60% of patients develop pathologic fractures over the course of their disease. Bone disease is a hallmark of MM, and myeloma bone disease differs from bone metastasis caused by other tumors. Although myeloma and other osteolytic metastases induce increased osteoclastic bone destruction, in contrast to other tumors, once myeloma tumor burden exceeds 50% in a local area, osteoblast activity is either severely depressed or absent. The basis for this severe imbalance between increased osteoclastic bone resorption and decreased bone formation has been the topic of intensive investigation over the last several years. These studies have helped to identify novel targets for treating myeloma bone disease and will be discussed in this chapter. PMID- 20014066 TI - RAD 001 (everolimus) prevents mTOR and Akt late re-activation in response to imatinib in chronic myeloid leukemia. AB - The mammalian target of rapamycin (mTOR) is one target of BCR-ABL fusion gene of chronic myeloid leukemia (CML). Moreover, it drives a compensatory route to Imatinib mesylate (IM) possibly involved in the progression of leukemic progenitors towards a drug-resistant phenotype. Accordingly, mTOR inhibitors are proposed for combined therapeutic strategies in CML. The major caveat in the use of mTOR inhibitors for cancer therapy comes from the induction of an mTOR phosphatidylinositol 3 kinase (PI3k) feedback loop driving the retrograde activation of Akt. Here we show that the rapamycin derivative RAD 001 (everolimus, Novartis Institutes for Biomedical Research) inhibits mTOR and, more importantly, revokes mTOR late re-activation in response to IM. RAD 001 interferes with the assembly of both mTOR complexes: mTORC1 and mTORC2. The inhibition of mTORC2 results in the de-phosphorylation of Akt at Ser(473) in the hydrophobic motif of C-terminal tail required for Akt full activation and precludes Akt re-phosphorylation in response to IM. Moreover, RAD 001-induced inhibition of Akt causes the de-phosphorylation of tuberous sclerosis tumor suppressor protein TSC2 at 14-3-3 binding sites, TSC2 release from 14-3-3 sigma (restoring its inhibitory function on mTORC1) and nuclear import (promoting the nuclear translocation of cyclin-dependent kinase [CDK] inhibitor p27(Kip1), the stabilization of p27(Kip1) ligand with CDK2, and the G(0)/G(1) arrest). RAD 001 cytotoxicity on cells not expressing the BCR-ABL fusion gene or its p210 protein tyrosine kinase (TK) activity suggests that the inhibition of normal hematopoiesis may represent a drug side effect. PMID- 20014068 TI - Resveratrol derivative, trans-3,5,4'-trimethoxystilbene, exerts antiangiogenic and vascular-disrupting effects in zebrafish through the downregulation of VEGFR2 and cell-cycle modulation. AB - Angiogenesis plays an important role in the development of neoplastic diseases such as cancer. Resveratrol and its derivatives exert antiangiogenic effects, but the mechanisms of their actions remain unclear. The aim of this study was to evaluate the antiangiogenic activity of resveratrol and its derivative trans 3,5,4'-trimethoxystilbene in vitro using human umbilical vein endothelial cells (HUVECs) and in vivo using transgenic zebrafish, and to clarify their mechanisms of action in zebrafish by gene expression analysis of the vascular endothelial growth factor (VEGF) receptor (VEGFR2/KDR) and cell-cycle analysis. trans-3,5,4' Trimethoxystilbene showed significantly more potent antiangiogenic activity than that of resveratrol in both assays. In zebrafish, trans-3,5,4'-trimethoxystilbene caused intersegmental vessel regression and downregulated VEGFR2 mRNA expression. Trans-3,5,4'-trimethoxystilbene also induced G2/M cell-cycle arrest, most specifically in endothelial cells of zebrafish embryos. We propose that the antiangiogenic and vascular-targeting activities of trans-3,5,4' trimethoxystilbene result from the downregulation of VEGFR2 expression and cell cycle arrest at G2/M phase. PMID- 20014069 TI - Human ES and iPS cells as cell sources for the treatment of Parkinson's disease: current state and problems. AB - Cell therapy using human embryonic stem cells (hESCs) is a promising therapeutic option for Parkinson's disease (PD), an incurable neurodegenerative disease. A prerequisite for clinical application of hESCs for PD is an efficient and strict differentiation of hESCs into midbrain dopamine (mDA) neuron-like cells, which would be directly translated into high effectiveness of the therapy with minimum risk of undesirable side effects. Due to fruitful efforts from many laboratories, a variety of strategies for improving efficiency of dopaminergic differentiation from hESCs have been developed, mostly by optimizing culture conditions, genetic modification, and modulating intracellular signaling pathways. The rapid advances in the fields of dopaminergic differentiation of hESCs, prevention of tumor formation, and establishment of safe human induced pluripotent stem cells (hiPSCs) would open the door to highly effective, tumor-free, and immune rejection-free cell therapy for PD in the near future. PMID- 20014070 TI - SAFB1's multiple functions in biological control-lots still to be done! AB - The examination of scaffold attachment factor B1 (SAFB1) and its multiple functions and tasks in cellular processes provides insight into its role in diseases, such as cancer. SAFB1 is a large multi-domain protein with well described functions in transcriptional repression, and RNA splicing. It is ubiquitously expressed, and has been shown to be important in numerous cellular processes including cell growth, stress response, and apoptosis. SAFB1 is part of a protein family with at least two other family members, SAFB2 and the SAFB-like transcriptional modulator SLTM. The goal of this prospect article is to summarize known functions of SAFB1, and its roles in cellular processes, but also to speculate on less well described, novel attributes of SAFB1, such as a potential role in chromatin organization. This timely review shows aspects of SAFB1, which are proving to have a complexity far greater than was previously thought. PMID- 20014071 TI - TremAn: a tool for measuring tremor frequency from video sequences. PMID- 20014072 TI - Deep brain stimulation in a patient with isolated mixed tremor. PMID- 20014073 TI - Depression and sexual adjustment following breast cancer in low-income Hispanic and non-Hispanic White women. AB - OBJECTIVE: Hispanics are the largest and fastest growing ethnic minority group in the United States and breast cancer is the most commonly diagnosed cancer in Hispanic women. However, Hispanics are underrepresented in the psychosocial breast cancer literature. METHODS: This study included 677 low-income women (425 Hispanic, 252 non-Hispanic White) enrolled in the Medi-Cal Breast and Cervical Cancer Treatment Program. Data were gathered through phone interviews conducted in English or Spanish 6 and 18 months following breast cancer diagnosis. We focus on three variables that the literature indicates are salient for breast cancer survivors: sexual function, body image and depression. RESULTS: Results of an ANCOVA indicated worse sexual function for Hispanic women, even after controlling for significant covariates. Hispanics reported significantly less sexual desire, greater difficulty relaxing and enjoying sex, and greater difficulty becoming sexually aroused and having orgasms than non-Hispanic White women. Both Hispanic and non-Hispanic White women endorsed a lack of sexual desire more frequently than problems with sexual function. Body image did not differ between Hispanic and non-Hispanic White women. In all, 38% of Hispanic and 48% of non-Hispanic White women scored above cut-off scores for depressive symptoms. While there was no ethnic difference in depressive symptoms, single women reported more depressive symptoms than partnered women. CONCLUSIONS: Findings suggest that low income breast cancer survivors may experience symptoms of depression more than a year following diagnosis, and that sexual dysfunction may be particularly salient for low-income Hispanic women. PMID- 20014074 TI - Trajectories of psychological distress among Chinese women diagnosed with breast cancer. AB - BACKGROUND: The distinct trajectories of psychological distress over the first year of the diagnosis with breast cancer (BC) and its determinants have not been explored. METHODS: 285 of 405 Chinese women receiving surgery for BC were assessed at 5-day, 1-month, 4-month, and 8-month post-surgery on measures of psychological distress, optimism, treatment decision-making (TDM) difficulties, satisfaction with treatment outcome, satisfaction with medical consultation, and physical symptom distress. Latent growth mixture modelling identified trajectories of psychological response to BC. Multinominal logistic regression compared TDM difficulties, satisfaction with treatment outcome, satisfaction with medical consultation, optimism, and physical symptom distress, by distress pattern adjusted for age, education, employment status, and stage of disease. RESULTS: Four distinct trajectories of distress were identified, namely, resilience (66%), chronic distress (15%), recovered (12%), and delayed-recovery (7%). TDM difficulties, optimism, satisfaction with consultation, and physical symptom distress predicted distress trajectories. Psychologically resilient women had less physical symptom distress at early post-surgery compared with women with other distress patterns. Compared with the resilient group, women in the recovered or chronic distress groups experienced greater TDM difficulties, whereas women in the delayed-recovery group reported greater dissatisfaction with the initial medical consultation. Women in the chronic distress group reported greater pessimistic outlook. CONCLUSION: Optimism and better early post-operative treatment outcomes predicted resilience to distress. Pre-operative interventions helping women to establish a realistic expectation of treatment outcome may minimize disappointment with treatment outcome and resultant distress, whereas post-operative rehabilitation should focus on symptom management. PMID- 20014075 TI - The development and impact of insomnia on cancer survivors: a qualitative analysis. AB - OBJECTIVES: To conduct the first qualitative analysis of the development and impact of insomnia on a cohort of cancer survivors. METHODS: Twenty-one cancer survivors with a history of chronic insomnia contributed to four focus groups held at the University of Glasgow Sleep Research Centre. Participants' perceptions of the onset, evolution and effects of insomnia were elicited and qualitatively explored using content analysis. RESULTS: Most participants reported insomnia onset following cancer diagnosis. Participants who had a pre existing insomnia reported that cancer diagnosis significantly aggravated their sleep complaint. Active cancer treatment was a major contributor to poor sleep quality due to the disruption of normal daily routines. This poor sleep pattern became persistent once active treatment had ceased and participants reported becoming particularly concerned about their sleep when they were discharged into follow-up cancer care. The impact of insomnia was significant for all participants in the study and six major areas emerged as being particularly affected; mood, physical health, relationships, sleep quality, sleep-related behaviour and cognition. CONCLUSIONS: The majority of cancer survivors in this study developed disturbed sleep as a result of cancer diagnosis and their sleep disruption was exacerbated by active cancer treatment. Insomnia also had a significant impact upon quality of life and these effects persisted long beyond the cessation of active anti-cancer therapy. Early identification of insomnia symptoms in cancer care settings must be a priority to ensure that sleep disturbance is not overlooked or poorly managed. PMID- 20014076 TI - Field evaluation of vaporised ethyl formate and carbon dioxide for fumigation of stored wheat. AB - BACKGROUND: Vapormate is a cylinderised non-flammable mixture of ethyl formate (16.7% by weight) and carbon dioxide (CO(2)) that has been developed as a rapid fumigant of stored grain. Four field trials were undertaken on wheat in 50 t farm silos to demonstrate the feasibility of dynamic application. To assess treatment efficacy, each trial tested mixed stages of Rhyzopertha dominica F. (>11,000), Tribolium castaneum (Herbst.) (>1500) and Sitophilus oryzae (L.) (>13,000) in mesh cages positioned through the centre of the grain bulk and on the grain surface. Ethyl formate and CO(2) concentrations were measured in the silo during fumigation and in ambient air outside the 6 m fumigation zone. Application rates of 420, 660 and 940 g m(-3) of ethyl formate/CO(2) formulation and exposure times of 24, 3 and 72 h, respectively, were examined using wheat of 10.4-11.7% moisture content and grain temperatures between 2 and 32 degrees C. RESULTS: All life stages of R. dominica and T. castaneum were fully controlled under all conditions tested, and mortality of all life stages of S. oryzae was greater than 98%. CONCLUSION: Dynamic application of vaporised ethyl formate and CO(2) to 50 t silos proved safe to operators and rapidly effective against stored-grain insects on cold to warm grain. PMID- 20014077 TI - A new highly selective and sensitive assay for fluorescence imaging of *OH in living cells: effectively avoiding the interference of peroxynitrite. AB - A new nonredox fluorescent probe to realize the imaging of hydroxyl radicals (*OH) in living cells was designed and synthesized. The structure comprised the fluorescent dye boron dipyrromethene (BDP) and a 2,2,6,6-tetramethyl-1 piperidinoxyl (TEMPO) unit. This probe could rapidly respond to *OH with a detection limit of 18 pM, and it possessed superior photostability and pH insensitivity. Other reactive oxygen species (ROS) and relevant intracellular components did not interfere. In particular, the important problem of ONOO(-) interference was efficiently avoided. An MTT assay proved that the probe was not very cytotoxic. The probe could penetrate into intact cell membranes to selectively detect intracellular *OH without causing cellular damage in living mice macrophages, normal human liver cells. and human hepatoma cells. These advantageous characteristics make the fluorescent probe potentially useful as a new candidate to detect *OH in broad biosystems. PMID- 20014078 TI - Applications of allylsamarium bromide as a grignard reagent and a single-electron transfer reagent in the one-pot synthesis of dienes and trienes. AB - The utility of allylsamarium bromide, both as a nucleophilic reagent and a single electron transfer (SET) reagent in the reaction of alpha-halo, gamma-halo alpha,beta-unsaturated ketones and esters with allylsamarium bromide is reported for the first time in this paper. From a synthetic point of view, a general, efficient and experimentally simple one-pot method for the preparation of 1,4 dienes and trienes is developed. A possible mechanism of the transformation is proposed. PMID- 20014079 TI - Biological activity of extracellular matrix-associated BMP-2. AB - The critical requirement for matrix-associated bone morphogenetic proteins (BMPs) during induction of bone formation in vivo has long been recognized. However, the role of extracellular matrix (ECM) physisorbed BMPs in inducing the differentiation of resident mesenchymal stem cells into osteoblasts has been ill defined. We therefore used BMP-responsive C2C12s to study the biological activity of collagen type I physisorbed BMP-2. Fibrillar collagen type I scaffolds were loaded with 75 ng BMP-2/microg collagen. Under cell culture conditions, 40% of loaded (125)I-labelled BMP-2 was released within 24 h, whereas the remaining BMP 2 was stably physisorbed for > 7 days. Using these systems suggested that physisorbed BMP-2 is more active than diffusible BMP-2. Thus, the current clinical practice of immobilizing BMPs on collagen type I scaffolds not only prolongs local delivery of the morphogen but could also enhance biological activity at the cellular level. PMID- 20014080 TI - Synthesis of a metal-organic framework material, iron terephthalate, by ultrasound, microwave, and conventional electric heating: a kinetic study. AB - A metal-organic framework material named MIL-53(Fe), iron terephthalate, has been synthesized sovothermally at a relatively low temperature by not only conventional electric (CE) heating, but also by irradiation under ultrasound (US) and microwave (MW) conditions to gain an understanding of the accelerated syntheses induced by US and MW. The kinetics for nucleation and crystal growth were analyzed by measuring the crystallinity of MIL-53(Fe) under various conditions. The nucleation and crystal growth rates were estimated from crystallization curves of the change in crystallinity with reaction time. The activation energies and pre-exponential factors were calculated from Arrhenius plots. It was confirmed that the rate of crystallization (both nucleation and crystal growth) decreases in the order US>MW>>CE, and that the accelerated syntheses under US and MW conditions are due to increased pre-exponential factors rather than decreased activation energies. It is suggested that physical effects such as hot spots are more important than chemical effects in the accelerated syntheses induced by US and MW irradiation. The syntheses were also conducted in two steps to understand quantitatively the acceleration induced by MW and it was found that the acceleration in crystal growth is more important than the acceleration in nucleation, even though both processes are accelerated by MW irradiation. PMID- 20014081 TI - Organocatalytic asymmetric synthesis of organophosphorus compounds. AB - 240 Years have passed since the discovery of elemental phosphorus. During that time organophosphorus chemistry has emerged as an interesting and exciting field of research. Recently organophosphorus chemistry has been raised to a new level. Organophosphorus compounds have found applications in asymmetric organocatalysis for the synthesis of optically active compounds of synthetic or biological importance. The aim of this review article is to present recent contributions to this developing field of chemistry and to point out synthetic advantages of methodologies developed so far. PMID- 20014082 TI - Chemoselectivity and the curious reactivity preferences of functional groups. AB - Achieving high levels of chemoselectivity has been the Achilles' heel of chemical synthesis. The excitement generated by the successful realization of chemoselective strategies underscores the painstaking efforts to define a set of conditions conducive to selection among the available reaction pathways. We discuss in this Review various aspects of chemoselectivity that have been addressed in a range of synthetic methods over the past decade. We have focused on the proposed mechanistic basis of the reactions under consideration in an attempt to categorize them and highlight the key concepts that have been emerging on the basis of these studies. Our overview of recent advances in chemoselective processes suggests that significant progress has been made, but a lot of challenges lie ahead. PMID- 20014083 TI - Binaphthyl-based dicationic peptoids with therapeutic potential. PMID- 20014084 TI - Multiple base-recognition sites in a biological nanopore: two heads are better than one. PMID- 20014085 TI - FRET-based activity biosensors to probe compartmentalized signaling. PMID- 20014086 TI - Haloacetamidine-based inactivators of protein arginine deiminase 4 (PAD4): evidence that general acid catalysis promotes efficient inactivation. AB - Dysregulated protein arginine deiminase (PAD) activity, particularly PAD4, has been suggested to play a role in the onset and progression of numerous human diseases, including rheumatoid arthritis (RA). Given the potential role of PAD4 in RA, we set out to develop inhibitors/inactivators that could be used to modulate PAD activity and disease progression. This effort led to the discovery of two mechanism-based inactivators, denoted F- and Cl-amidine, that inactivate PAD4 by the covalent modification of an active-site cysteine that is critical for catalysis. To gain further insights into the mechanism of inactivation by these compounds, the effect of pH on the rates of inactivation was determined. These results, combined with the results of solvent isotope effect and proton inventory studies, strongly suggest that the inactivation of PAD4 by F- and Cl-amidine proceeds by a multistep mechanism that involves the protonation and stabilization of the tetrahedral intermediate formed upon nucleophilic attack by the active site cysteine, that is, Cys645. Stabilization of this intermediate would help to drive the halide-displacement reaction, which results in the formation of a three membered sulfonium ring that ultimately collapses to form the inactivated enzyme. This finding-that protonation of the tetrahedral intermediate is important for enzyme inactivation-also suggests that, during catalysis, protonation of the analogous intermediate is required for efficient substrate turnover. PMID- 20014087 TI - Optimal configurations of "capped" beta-cyclodextrin dimers in water maximise hydrophobic association. AB - Circular dichroism analysis and proton NMR experiments revealed that solutions of 3-O-(2-methylnaphthyl)-beta-cyclodextrin form different dimer configurations. The exact nature of the dimer configurations were postulated to be of three types in which these capped cyclodextrins (CDs) are orientated in head-to-head and head-to tail arrangements. Here we show from detailed computer simulations and free energy calculations on the configurations that the head-to-head configuration in which the naphthyl groups are mutually inserted into each other's CD cavities is the most favoured configuration. This configuration optimises the hydrophobic association of the naphthyl aromatic groups and the ring cavities as well as forming the most inter-CD hydrogen bonds of the three configurations. PMID- 20014088 TI - Scaffold distributions in bioactive molecules, clinical trials compounds, and drugs. PMID- 20014089 TI - Organoplatinum(II) complexes with chromophore-acceptor dyad studied by ultrafast time-resolved absorption spectroscopy. PMID- 20014090 TI - Luminescence screening assays for the identification of sensitizers for lanthanides based on the controlled formation of ternary lanthanide complexes with DTPA-bisamide ligands. AB - The formation of ternary complexes of lanthanide-diethylenetriamine pentaacetic acid (DTPA)-bisamide complexes with different aromatic acids and their application in luminescent screening assays are presented. The europium complexes of DTPA-bisethylamide (BEA), DTPA-bisbutylamide (BUA), DTPA-bis(2-norbornyl) (NBA), and DTPA-bis(1-adamantyl) (ADA) have been isolated and the sensitization of emission upon 1:1 formation with aromatic acids has been studied by luminescence spectroscopy. The ternary complexes show stronger luminescence with picolinate (PCA) rather than phthalate (PTA) or benzoate (BZA), with the latter forming 1:2 complexes. Isophthalate and dipicolinate sensitizers do not show a 1:1 formation of the ternary complexes. Electrospray mass spectra show characteristic peaks that confirm the formation of the ternary complexes, and NMR spectroscopic studies demonstrated a conformational locking effect upon formation of the ternary complex. It is shown that europium complexes with bulkier amide arms provide complexes with stronger luminescence enhancement that is clearly attributable to the bulkiness of the amide arms. In a luminescence screening experiment, different substitutions on BZA and PCA were examined. Sensitizers with a long alkyl chain show greater luminescent enhancements than the nonsubstituted acids, which can be attributed to an "umbrella" effect of the alkyl chain that protects the europium luminescent center from the quenching of secondary high-energy vibrations, in particular O-H from water molecules. The same effect is presented for the quinoline derivatives as sensitizers. Quinoline derivatives with salicylate binding units were identified as the best sensitizers when combined with EuADA, which is even bulkier than EuNBA, with HQ-3COO showing a 50-fold enhancement of emission upon formation of 1:1 complexes. PMID- 20014091 TI - MoO3 in self-organized TiO2 nanotubes for enhanced photocatalytic activity. PMID- 20014092 TI - Chemistry of renewables. PMID- 20014093 TI - Electrical interaction between cardiomyocyte sheets separated by non cardiomyocyte sheets in heterogeneous tissues. AB - Electrical coupling between cardiomyocytes is important in synchronous beating and normal heart functions. Cardiomyocytes are also electrically coupled to non cardiomyocytes. The electrical interactions between cardiomyocytes and non cardiomyocytes, or those between separated cardiomyocytes, are important for normal heart function because abnormalities of the coupling and variation of the cell population induce pathological heart functions and arrhythmias. In this study the three-dimensional time course of the electrical interaction between two rat neonatal cardiomyocyte sheets separated by non-cardiomyocyte sheets was analysed by a multiple-electrode extracellular recording system. The two cardiomyocyte sheets separated by a single- or double-layered mouse fibroblast NIH3T3 cell sheet coupled electrically at 113 +/- 28 or 287 +/- 87 min after layering, respectively. The time course of the electrical coupling, when the single-layer NIH3T3 cell sheet was inserted, is similar to that of a layered cardiomyocyte sheet. Immunocytological analysis and dye transfer assay suggested the formation of gap junctions at heterocellular junctions of cardiomyocytes and NIH3T3 cells. On the other hand, when a double-layered NIH3T3 cell sheet was inserted, an incomplete electrical coupling of two cardiomyocyte sheets, including a conduction delay, was observed. The electrical coupling of cardiomyocyte sheets was completely blocked (conduction block) by insertion of a triple-layered NIH3T3 cell sheet, a communication-defective HeLa cell sheet or a Ca(2+)-antagonist LaCl(3)-treated cell sheet. These electrophysiological analyses of heterogeneously stacked cell sheets might provide insights into complex electrical conduction systems that resemble those of native or damaged heart and transplanted tissues. PMID- 20014094 TI - Calmin expression in embryos and the adult brain, and its regulation by all-trans retinoic acid. AB - The vitamin A metabolite, all-trans retinoic acid (atRA), is a regulator of nervous system development. Using a subtracted cDNA library constructed from neuroblastoma cells, the atRA-responsive gene calmin (Clmn) was identified (Merrill et al. [2004] Biol Chem 385:605-614). The Clmn transcript is detected very early in rat embryonic development and is sensitive to retinoid status. In vitamin A-deficient embryos, Clmn mRNA is dramatically down-regulated in the neuroepithelium adjacent to the somites, and this expression can be rescued with the addition of atRA. In embryonic day 18.5 embryos, CLMN is detected in regions where newly differentiated neurons are found, including the neural retina and the cortical plate; and in the adult brain, CLMN is most highly expressed in the neuron cell bodies of the hippocampus, cerebellum, and olfactory bulb. Thus, Clmn is sensitive to retinoid status during early gestational stages, and its expression is relegated to postmitotic neuronal cells in the adult rat brain. PMID- 20014095 TI - Relative clinical utility of three Child Symptom Inventory-4 scoring algorithms for differentiating children with autism spectrum disorder vs. attention-deficit hyperactivity disorder. AB - OBJECTIVE: The present study compared three separate Child Symptom Inventory-4 (CSI-4) scoring algorithms for differentiating children with autism spectrum disorder (ASD) from youngsters with attention-deficit/hyperactivity disorder (ADHD). METHOD: Parents/teachers completed the CSI-4, a DSM-IV-referenced rating scale, for 6 to 12-year-old clinical referrals with ASD (N = 186) and ADHD (N = 251). Algorithms were based on either all CSI-4 items (forward logistic regressions) or the 12 DSM-IV symptoms of pervasive developmental disorder (PDD) included in the CSI-4. RESULTS: ROC analyses indicated generally good to excellent values for area under the curve, sensitivity, specificity, and positive predictive power. Algorithms for parent ratings were superior to teacher ratings. The algorithm based solely on PDD symptoms evidenced the greatest generalizability. CONCLUSION: Although algorithms generated from regression analyses produced greater clinical utility for specific samples, the PDD-based algorithm resulted in greater stability across samples. PMID- 20014096 TI - Maintenance of pluripotency in mouse embryonic stem cells cultivated in stirred microcarrier cultures. AB - The development of efficient and reproducible culture systems for embryonic stem (ES) cells is an essential pre-requisite for regenerative medicine. Culture scale up ensuring maintenance of cell pluripotency is a central issue, because large amounts of pluripotent cells must be generated to warrant that differentiated cells deriving thereof are transplanted in great amounts and survive the procedure. This study aimed to develop a robust scalable cell expansion system, using a murine embryonic stem cell line that is feeder-dependent and adapted to serum-free medium, thus representing a more realistic model for human ES cells. We showed that high concentrations of murine ES cells can be obtained in stirred microcarrier-based spinner cultures, with a 10-fold concentration of cells per volume of medium and a 5-fold greater cell concentration per surface area, as compared to static cultures. No differences in terms of pluripotency and differentiation capability were observed between cells grown in traditional static systems and cells that were replated onto the traditional system after being expanded on microcarriers in the stirred system. This was verified by morphological analyses, quantification of cells expressing important pluripotency markers (Oct-4, SSEA-1, and SOX2), karyotype profile, and the ability to form embryoid bodies with similar sizes, and maintaining their intrinsic ability to differentiate into all three germ layers. PMID- 20014097 TI - The formation of the superior and jugular ganglia: insights into the generation of sensory neurons by the neural crest. AB - The superior and jugular ganglia (S/JG) are the proximal ganglia of the IXth and Xth cranial nerves and the sensory neurons of these ganglia are neural crest derived. However, it has been unclear the extent to which their differentiation resembles that of the Dorsal Root Ganglia (DRGs). In the DRGs, neural crest cells undergo neuronal differentiation just after the onset of migration and there is evidence suggesting that these cells are pre-specified towards a sensory fate. We have analysed sensory neuronal differentiation in the S/JG. We show, in keeping with previous studies, that neuronal differentiation initiates long after the cessation of neural crest migration. We also find no evidence for the existence of migratory neural crest cells pre-specified towards a sensory phenotype prior to ganglion formation. Rather our results suggest that sensory neuronal differentiation in the S/JG is the result of localised spatiotemporal cues. PMID- 20014098 TI - The tight junction component Claudin E is required for zebrafish epiboly. AB - Zebrafish epiboly results in the thinning and spreading of the blastoderm to cover the yolk cell and close the blastopore. The extra-embryonic yolk syncytial layer (YSL) tows the blastoderm vegetally during epiboly by means of its tight junction attachments to the enveloping layer (EVL). Claudins are the major transmembrane protein components of tight junctions. Here, we focus on the function of Claudin E (Cldne), which is expressed specifically in the EVL. Morpholino knock-down of cldne produced a highly penetrant epiboly delay. Our analysis suggested that the EVL margin, which is attached to the YSL, was under reduced tension in morphant embryos. We propose that local variation in the strength of EVL-YSL attachment in morphant embryos resulted in slow and uneven advancement of the EVL and blastoderm. Our work is the first to demonstrate that Claudins are important for zebrafish epiboly. PMID- 20014099 TI - High-throughput screening techniques for rapid PEG-based precipitation of IgG4 mAb from clarified cell culture supernatant. AB - Locating optimal protein precipitation conditions for complex biological feed materials is problematic. This article describes the application of a series of high-throughput platforms for the rapid identification and selection of conditions for the precipitation of an IgG(4) monoclonal antibody (mAb) from a complex feedstock using only microliter quantities of material. The approach uses 96-microwell filter plates combined with high-throughput analytical methods and a method for well volume determination for product quantification. The low material, time and resource requirements facilitated the use of a full factorial Design of Experiments (DoE) for the rapid investigation into how critical parameters impact the IgG(4) precipitation. To aid the DoE, a set of preliminary range-finding studies were conducted first. Data collected through this approach describing Polyethylene Glycol (PEG) precipitation of the IgG(4) as a function of mAb concentration, precipitant concentration, and pH are presented. Response surface diagrams were used to explore interactions between parameters and to inform selection of the most favorable conditions for maximum yield and purification. PEG concentrations required for maximum yield and purity were dependant on the IgG(4) concentration; however, concentrations of 14 to 20% w/v, pH 6.5, gave optimal levels of yield and purity. Application of the high throughput approach enabled 1,155 conditions to be examined with less than 1 g of material. The level of insights gained over such a short time frame is indicative of the power of microwell experimentation in allowing the rapid identification of appropriate processing conditions for key bioprocess operations. PMID- 20014101 TI - Mouse ZAR1-like (XM_359149) colocalizes with mRNA processing components and its dominant-negative mutant caused two-cell-stage embryonic arrest. AB - Maternal effect genes and encoding proteins are necessary for nuclear reprogramming and zygotic genome activation. However, the mechanisms that mediate these functions are largely unknown. Here we identified XM_359149, a Zar1-like gene that is predominantly expressed in oocytes and zygotes, which we designated Zar1-like (Zar1l). ZAR1L-EGFP formed multiple cytoplasmic foci in late two-cell stage embryos. Expression of the ZAR1L C-terminus induced two-cell-stage embryonic arrest, accompanied with abnormal methylation of histone H3K4me2/3 and H3K9me2/3, and marked down-regulation of a group of chromatin modification factors including Dppa2, Dppa4, and Piwil2. When ectopically expressed in somatic cells, ZAR1L colocalized with P-body components including EIF2C1(AGO1), EIF2C2(AGO2), DDX6 and LSM14A, and germline-specific chromatoid body components including PIWIL1, PIWIL2, and LIN28. ZAR1L colocalized with ZAR1 and interacted with human LIN28. Our data suggest that ZAR1L and ZAR1 may comprise a novel family of processing-body/chromatoid-body components that potentially function as RNA regulators in early embryos. PMID- 20014102 TI - Wnt signaling has different temporal roles during retinal development. AB - Differentiation of neural retinal precursor (NRP) cells in vertebrates follows an established order of cell-fate determination associated with exit from the cell cycle. Wnt signaling regulates cell cycle in colon carcinoma cells and has been implicated in different aspects of retinal development in various species. To better understand the biological roles of Wnt in the developing retina, we have used a transgenic and pharmacological approach to manipulate the Wnt signaling pathway during retinal development in medaka embryos. With the use of both approaches, we observed that during the early phase of retinal development Wnt signaling regulated cell cycle progression, proliferation, apoptosis, and differentiation of NRP cells. However, during later phases of retinal development, proliferation and apoptosis were not affected by manipulation of Wnt signaling. Instead, Wnt regulated Vsx1 expression, but not the expression of other retinal cell markers tested. Thus, the response of NRP cells to Wnt signaling is stage-dependent. PMID- 20014100 TI - Wnt modulators in the biotech pipeline. AB - The purpose of this review is to provide a better understanding for the LRP co receptor-mediated Wnt pathway signaling. Using proteomics, we have also subdivided the LRP receptor family into six sub-families, encompassing the twelve family members. This review includes a discussion of proteins containing a cystine-knot protein motif (i.e., Sclerostin, Dan, Sostdc1, Vwf, Norrin, Pdgf, Mucin) and discusses how this motif plays a role in mediating Wnt signaling through interactions with LRP. PMID- 20014103 TI - Enhancement of cell recovery for dissociated human embryonic stem cells after cryopreservation. AB - Due to widespread applications of human embryonic stem (hES) cells, it is essential to establish effective protocols for cryopreservation and subsequent culture of hES cells to improve cell recovery. We have developed a new protocol for cryopreservation of dissociated hES cells and subsequent culture. We examined the effects of new formula of freezing solution containing 7.5% dimethylsulfoxide (DMSO) (v/v %) and 2.5% polyethylene glycol (PEG) (w/v %) on cell survival and recovery of hES cells after cryopreservation, and further investigated the role of the combination of Rho-associated kinase (ROCK) inhibitor and p53 inhibitor on cell recovery during the subsequent culture. Compared with the conventional slow freezing method which uses 10% DMSO as a freezing solution and then cultured in the presence of ROCK inhibitor at the first day of culture, we found out that hES cell recovery was significantly enhanced by around 30 % (P < 0.05) by the new freezing solution. Moreover, at the first day of post-thaw culture, the presence of 10 microM ROCK inhibitor (Y-27632) and 1 microM pifithrin-mu together further significantly improved cell recovery by around 20% (P < 0.05) either for feeder dependent or feeder-independent culture. hES cells remained their undifferentiated status after using this novel protocol for cryopreservation and subsequent culture. Furthermore, this protocol is a scalable cryopreservation method for handling large quantities of hES cells. PMID- 20014105 TI - Comparison of extraembryonic expression of Eomes and Cdx2 in pregastrulation chick and mouse embryo unveils regulatory changes along evolution. AB - In the mouse blastocyst, Eomes and Cdx2 are critical for establishing the trophoectoderm, the precursor of the placenta. To better understand how the trophoectoderm lineage arose in mammals during evolution, we examined the expression of their orthologues in the pregastrulation chick embryo and found that, while both genes are expressed in extraembryonic tissues, their temporal pattern of expression differs from what occurs in mouse. Moreover, we failed to detect expression of other genes specific from the mouse trophoectoderm in extraembryonic regions of the chick. Also unlike the mouse, chick Eomes is expressed in primordial germ cells. Finally, conserved noncoding elements in the Eomes genomic region are unable to drive trophoectoderm restricted expression in the mouse blastocyst, but do so in conserved sites of expression such as the forebrain. These results suggest that critical changes in the gene regulatory networks controlling extraembryonic development accompanied the appearance of the trophoectoderm in mammals. PMID- 20014106 TI - Science communications: publishing a scientific paper. AB - Publications are the lifeblood of academic science. They are essential to the scientific community as records of research completed, building blocks for new research, and templates for new ways of thinking. Publications are also essential for individual job security. Frequently they are considered a gauge of productivity that is taken under heavy consideration for hiring, promotion, and funding decisions. Consequently, completed research is of little value unless preserved in publication. Yet publishing can be a challenging prospect, particularly in the face of abundant competition. Here, five panelists, John Fallon, Ph.D., Min Han, Ph.D., Janet Rossant, Ph.D., Cliff Tabin, Ph.D., and Yoshiko Takahashi, Ph.D., offer their advice on manuscript preparation and negotiating the publishing process. PMID- 20014104 TI - pHMA, a pH-sensitive GFP reporter for cell engulfment, in Drosophila embryos, tissues, and cells. AB - Engulfment of apoptotic cells by phagocytosis ensures the removal of unwanted and defective cells. We developed a genetically encoded marker for cell engulfment, pHMA, which consists of the pH-Sensitive derivative of GFP, pHluorin, fused to the actin-binding domain of Moesin. In healthy cells of Drosophila embryos and cultured cells, pHMA resides at the cell cortex. In dying cells, pHMA loses its cortical localization and reports a modest decrease in pH. In embryos, the dying cells lose their apical contacts, then move basally and are ultimately engulfed by neighboring cells or macrophages. The cell corpse material is strongly acidified soon after engulfment and persists in the phagocytic cell for several hours. Changes in the pHMA signal correlate well with increases or decreases in apoptosis. These data show that pHMA is a useful reporter for cell engulfment and can be used in screening for mutations that affect cell engulfment. PMID- 20014107 TI - Active and inactive metabolic pathways in tumor spheroids: determination by GC MS. AB - Active metabolic pathways in three-dimensional cancer-cell cultures are potential chemotherapeutic targets that would be effective throughout tumors. Chaotic vasculature creates cellular regions in tumors with distinct metabolic behavior that are only present in aggregate cell masses. To quantify cancer cell metabolism, transformed mouse fibroblasts were grown as spheroids and fed isotopically labeled culture medium. Metabolite uptake and production rates were measured as functions of time. Gas chromatography-mass spectrometry was used to quantify the extent of labeling on amino acids present in cytoplasmic extracts. The labeling pattern identified several active and inactive metabolic pathways: Glutaminolysis was found to be active, and malic enzyme and gluconeogenesis were inactive. Transformed cells in spheroids were also found to actively synthesize serine, cysteine, alanine, aspartate, glutamate, and proline; and not synthesize glutamine. The activities of these pathways suggest that cancer cells consume glutamine for biosynthesis and not to provide cellular energy. Determining active metabolic pathways indicates how cells direct carbon flow and may lead to the discovery of novel molecular targets for anticancer therapy. PMID- 20014108 TI - Feed development for fed-batch CHO production process by semisteady state analysis. AB - Semisteady state cultures are useful for studying cell physiology and facilitating media development. Two semisteady states with a viable cell density of 5.5 million cells/mL were obtained in CHO cell cultures and compared with a fed-batch mode control. In the first semisteady state, the culture was maintained at 5 mM glucose and 0.5 mM glutamine. The second condition had threefold higher concentrations of both nutrients, which led to a 10% increase in lactate production, a 78% increase in ammonia production, and a 30% reduction in cell growth rate. The differences between the two semisteady states indicate that maintaining relatively low levels of glucose and glutamine can reduce the production of lactate and ammonia. Specific amino acid production and consumption indicated further metabolic differences between the two semisteady states and fed batch mode. The results from this experiment shed light in the feeding strategy for a fed-batch process and feed medium enhancement. The fed-batch process utilizes a feeding strategy whereby the feed added was based on glucose levels in the bioreactor. To evaluate if a fixed feed strategy would improve robustness and process consistency, two alternative feeding strategies were implemented. A constant volume feed of 30% or 40% of the initial culture volume fed over the course of cell culture was evaluated. The results indicate that a constant volumetric-based feed can be more beneficial than a glucose-based feeding strategy. This study demonstrated the applicability of analyzing CHO cultures in semisteady state for feed enhancement and continuous process improvement. PMID- 20014109 TI - Variation and context of yawns in captive chimpanzees (Pan troglodytes). AB - Primate yawns are usually categorized according to context (e.g. as a threat, anxious, or rest yawn), but there has been little consideration of whether these yawns are best regarded as a unitary behavior that only differs with respect to the context in which it is observed. This study examined the context and precise morphology of yawns in a group of 11 captive chimpanzees. Focal video sampling was used to describe the morphology and intensity of 124 yawns using ChimpFACS, a system for coding facial movements. Two distinct forms of yawn were identified, a full yawn and a yawn which is modified by additional actions that reduce the mouth aperture. These modified yawns may indicate some degree of voluntary control over facial movement in chimpanzees and, consequently, multiple functions of yawning according to context. To assess context effects, mean activity levels (resting, locomotion, and grooming) and scratching rates were compared one minute before and after each yawn. Locomotion was significantly increased following both types of yawn, whereas scratching rates significantly increased following modified yawns but decreased following full yawns. In terms of individual differences, males did not yawn more than females, although male yawns were of higher intensity, both in the degree of mouth opening and in the amount of associated head movement. These data indicate that yawning is associated with a change in activity levels in chimpanzees, but only modified yawns may be related to increased arousal. Different types of yawn can therefore be differentiated at the morphological level as well as context level. PMID- 20014110 TI - Motor laterality asymmetry and nonmotor symptoms in Parkinson's disease. AB - BACKGROUND: In patients with Parkinson's disease (PD), asymmetric motor signs provide an interesting model to evaluate whether asymmetric nigrostriatal degeneration can affect neuropsychological function and other nonmotor symptoms (NMS). This study was designed to evaluate the predominant laterality of motor symptoms and its relationship with cognition and other NMS in idiopathic PD. METHODS: Nationwide, longitudinal, and multicenter study (ELEP Registry) using outpatients with PD. Left PD (LPD) and right PD (RPD) was defined based on the motor signs on the SCOPA-motor scale. To include the clinical spectrum of asymmetric PD patients, we considered two groups of patients with mild-moderate and extreme asymmetry. Predominant LPD or RPD with mild-moderate versus extreme asymmetry were compared using the following scales: cognition, psychosis (Parkinson Psychosis Rating Scale), anxiety/depression, sleep (and autonomic dysfunction at baseline and 1 year later. Nonparametric tests were used for comparison. RESULTS: One hundred forty-nine PD patients (74 RPD and 75 LPD) with mild-moderate asymmetry and 90 (47 RPD and 43 LPD) with extreme asymmetry and a mean age of 64.5 (10.4) years were included. Extreme RPD had higher Parkinson Psychosis Rating Scale scores over time (P = 0.005) compared with LPD, but no significant differences were observed between LPD and RPD in terms of other NMS. CONCLUSIONS: These findings suggest that damage to left-hemisphere plays a disproportionately greater role in PD-related psychosis over time. In contrast, motor laterality does not consistently affect other NMS, suggesting that NMS are related to a more widespread brain disorder. PMID- 20014111 TI - A survey of elephant husbandry and foot health in North American zoos. AB - The foot health of elephants in human care is a longstanding concern. In 2001, the AZA Standards for Elephant Management and Care were published recommending husbandry to improve foot health. This article reports the results of a 2006 survey: basic statistics describing facility, husbandry, and foot health attributes are reported and relationships among variables are investigated. Median area available to elephants exceeded Standard recommendations (755 ft(2) per elephant indoor and 10,000 ft(2) outdoor). Concrete makes up 69% of indoor area and natural substrates account for 85% of outdzoor area. Elephants in AZA facilities received an average of 45.5 min/day of exercise, and facilities with a structured exercise plan provided significantly more exercise than did facilities without a structured exercise plan (z=-2.522, P=0.012). Enrichment is important to psychological health and may also stimulate activity beneficial to foot health; 95% of institutions had a structured enrichment program. Preventative foot care was nearly universal, and 100% of facilities performed routine nail and pad trimming. However, foot pathology has not been eradicated; 33% of institutions reported at least one pathology in the previous year. This study found a strong inverse relationship between foot pathology and exercise (chi(2)(3)=24.34, P<0.001). Younger herds were less likely to have a member diagnosed with arthritis (chi(2)(1)=8.90, P=0.003). Lameness was unrelated to age or pathology, and only the presence of arthritis explained lameness (z=-7.81, P<0.001). African elephants seemed to experience lower rates of foot pathology and arthritis than Asian elephants; however, this was explained by differences in age. PMID- 20014112 TI - Differential neuropsychological profiles in Parkinsonian patients with or without vascular lesions. AB - The purpose of this study is to compare the neuropsychological profile of patients affected by parkinsonism and vascular lesions to that in patients with PD alone (PD) and to evaluate whether the brain vascular lesion load is associated with neuropsychological variables. Thirty-six nondemented patients with parkinsonism were divided into 3 groups of 12 patients each, according to both clinical history and the presence of brain vascular lesions and/or dopaminergic denervation as revealed by magnetic resonance and dopamine transporter imaging, respectively. The first group had vascular lesions without dopaminergic denervation (VP group); the second group had vascular lesions and dopaminergic denervation (DD) (VP+DD group); and the third group consisted of patients with dopaminergic denervation (PD group) without vascular lesions. All patients underwent neurological and neuropsychological assessments. The groups differed in disease duration, age at onset, and cerebrovascular risk factors. The VP and VP+DD groups performed worse than the PD group on frontal/executive tasks. Regardless of the presence of dopaminergic denervation, cerebrovascular lesions in hemispheric white matter, basal ganglia, and cerebellum have an important effect in determining early onset and severity of cognitive impairment in patients with parkinsonism. PMID- 20014113 TI - Novel PANK2 gene mutations in korean patient with pantothenate kinase-associated neurodegeneration presenting unilateral dystonic tremor. PMID- 20014114 TI - Decreased survival of subjects with elevated liver function tests during a 28 year follow-up. AB - The long-term survival of subjects with nonalcoholic fatty liver disease (NAFLD) in comparison with both individuals with elevated transaminases attributable to other causes and the general poulation is poorly characterized. This study was undertaken to determine the frequency of NAFLD in a cohort of subjects who underwent liver biopsy from 1980 to 1984 because of elevated liver enzymes, and to assess mortality among subjects with NAFLD in comparison with the general Swedish population. The 256 subjects (61% men) had a mean age of 45 +/- 12 years at the inclusion. Liver biopsies were blindly scored for NAFLD and nonalcoholic steatohepatitis (NASH). Causes of death were ascertained from the national Swedish Cause of Death Registry. Fatty liver was detected in 143 of the 256 subjects, including 25 (10%) with alcoholic fatty liver disease and 118 (46%) exhibiting NAFLD. Of those, 51 (20%) were classified as NASH and 67 (26%) as nonalcoholic bland steatosis. Cirrhosis was present in 9% at inclusion. During the follow-up period, 113 (44%) of the total population and 47 (40%) of the 118 subjects diagnosed with NAFLD died. Of the 113 deaths, 37 were of cardiovascular disease and 16 of liver diseases. Compared with the total Swedish population, adjusted for sex, age, and calendar period, subjects with NAFLD exhibited a 69% increased mortality (standardized mortality ratio [SMR] = 1.69; 95% confidence interval [CI], 1.24-2.25); subjects with bland steatosis, a 55% increase (SMR, 1.55; 95% CI, 0.98-2.32; P = 0.062); and subjects with NASH, 86% (SMR, 1.86; 95% CI, 1.19-2.76; P = 0.007). CONCLUSION: Patients with NASH are at increased risk of death compared with the general population. Liver disease is the third most common cause of death among patients with NAFLD. PMID- 20014115 TI - BDNF levels are not related with levodopa-induced dyskinesias in MPTP monkeys. AB - Levodopa-induced dyskinesias (LIDs) are frequent in parkinsonian patients and may result from an aberrant plasticity. Brain-derived neurotrophic factor (BDNF) represents a likely candidate to subserve neuroadaptive processes encountered in LIDs. We compared striatal BDNF levels measured by ELISA in levodopa-treated 1 methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) monkeys having developed LIDs compared with animals where LIDs were prevented by the addition of CI-1041 (NR1A/2B NMDA receptor antagonist) or low doses of cabergoline (dopamine D2 receptor agonist). We observed reduced striatal BDNF concentrations in levodopa treated MPTP monkeys with or without LIDs, suggesting that levodopa treatment is associated with reduced striatal BDNF levels and is independent of dyskinesias. PMID- 20014116 TI - Baroreflex sensitivity and power spectral analysis during autonomic testing in different extrapyramidal syndromes. AB - Autonomic dysfunction has been frequently demonstrated in patients with extrapyramidal diseases by cardiovascular autonomic testing. In addition to classical testing, we applied the more detailed baroreflex and spectral analysis on three traditional cardiovascular tests in this study to get additional information on autonomic outflow. We recorded continuously blood pressure, electrocardiogram, and respiration in 35 patients with multiple system atrophy, 32 patients with progressive supranuclear palsy, 46 patients with idiopathic Parkinson's disease and in 27 corresponding healthy subjects during cardiovascular autonomic testing (metronomic breathing, Valsalva manoeuvre, head up tilt). Baroreflex and spectral analyses were performed by using trigonometric regressive spectral analysis between and during the manoeuvres. Consistent with previous interpretations, our data showed an increase of sympathetic activity in head-up tilt and Valsalva test in healthy controls. This sympathetic activity was significantly decreased in patients with typical and atypical Parkinson syndromes. Significant modulation of baroreflex activity could be observed especially during metronomic breathing; again it was significantly lower in all patient groups. Baroreflex and spectral parameters could not only differentiate between patients and healthy controls, but also differentiate between clinically symptomatic (with autonomic dysfunction as eg. orthostatic hypotension) and asymptomatic patients. In conclusion, our approach allows the evaluation of autonomic variability during short and nonstationary periods of time and may constitute a useful advance in the assessment of autonomic function in both physiological and pathological conditions. PMID- 20014117 TI - Clinical measures of dysarthria in Friedreich Ataxia. AB - Dysarthria in Friedreich Ataxia (FA) is difficult to quantify. This study evaluated a series of performance measures for speech in 22 patients with genetically confirmed FA and 16 age-matched controls. Tests included the PATA examination, the PATAKA examination, the Oral Motor component of the Boston Aphasia examination, the Boston Cookie Theft description task, and the Assessment of Intelligibility of Dysarthric Speech. All measures, except the Cookie theft description task, demonstrated significantly lower scores for patients with FA when compared with controls and correlated with measures of disease progression. Thus, four of five measures capture speech dysfunction in FA and may provide feasible, inexpensive, quantitative testing for therapeutic monitoring in FA. PMID- 20014118 TI - Minocycline 1-year therapy in multiple-system-atrophy: effect on clinical symptoms and [(11)C] (R)-PK11195 PET (MEMSA-trial). AB - The aim of the study was to investigate the efficacy of the antibiotic minocycline as a drug treatment in patients with Multiple-System-Atrophy Parkinson-type (MSA-P). Sixty-three patients were randomized to minocycline 200 mg/d (n = 32) or a matching placebo (n = 31). The primary outcome variable was the change in the value of the motor score of the Unified Multiple-System-Atrophy Rating-Scale (UMSARSII) from baseline to 48 weeks. Secondary outcome variables included subscores and individual Parkinsonian symptoms as determined by the UMSARS and the Unified-Parkinson's-Disease Rating-Scale (UPDRS). Health-related quality of life (HrQoL) was assessed using the EQ-5D and SF-12. "Progression rate" was assumed to be reflected in the change in motor function over 48 weeks. At 24 weeks and 48 weeks of follow-up, there was a significant deterioration in motor scores in both groups, but neither the change in UMSARSII nor in UPDRSIII differed significantly between treatment groups, i.e. "progression rate" was considered to be similar in both treatment arms. HrQoL did not differ among the two treatment arms. In a small subgroup of patients (n = 8; minocycline = 3, placebo = 5)[(11)C](R)-PK11195-PET was performed. The three patients in the minocycline group had an attenuated mean increase in microglial activation as compared to the placebo group (P = 0.07) and in two of them individually showed decreased [11C](R)-PK11195 binding actually decreased. These preliminary PET-data suggest that minocycline may interfere with microglial activation. The relevance of this observation requires further investigation. This prospective, 48 week, randomized, double-blind, multinational study failed to show a clinical effect of minocycline on symptom severity as assessed by clinical motor function. PMID- 20014119 TI - Proceedings from the 2009 genetic syndromes of the Ras/MAPK pathway: From bedside to bench and back. AB - The RASopathies are a group of genetic syndromes caused by germline mutations in genes that encode components of the Ras/mitogen-activated protein kinase (MAPK) pathway. Some of these syndromes are neurofibromatosis type 1, Noonan syndrome, Costello syndrome, cardio-facio-cutaneous syndrome, LEOPARD syndrome and Legius syndrome. Their common underlying pathogenetic mechanism brings about significant overlap in phenotypic features and includes craniofacial dysmorphology, cardiac, cutaneous, musculoskeletal, GI and ocular abnormalities, and a predisposition to cancer. The proceedings from the symposium "Genetic Syndromes of the Ras/MAPK Pathway: From Bedside to Bench and Back" chronicle the timely and typical research symposium which brought together clinicians, basic scientists, physician scientists, advocate leaders, trainees, students and individuals with Ras syndromes and their families. The goals, to discuss basic science and clinical issues, to set forth a solid framework for future research, to direct translational applications towards therapy and to set forth best practices for individuals with RASopathies were successfully meet with a commitment to begin to move towards clinical trials. PMID- 20014121 TI - Fowler syndrome-a clinical, radiological, and pathological study of 14 cases. AB - We report on 14 fetuses from 10 families with the autosomal recessive syndrome of proliferative vasculopathy and hydranencephaly-hydrocephaly (Fowler syndrome). In four families sibs were affected and in six the parents were consanguineous. Antenatal ultrasonography showed hydrocephaly in all except two fetuses, but hydranencephaly was diagnosed in only one case. Postural abnormalities were seen in 10 fetuses and structural brain abnormalities were suspected in 3. At autopsy the cerebral cortex appeared as a translucent membranous structure (hydranencephaly) in most fetuses. However, in one case, the ventricles were dilated but the cortical mantle was relatively well preserved. Histology of the brain showed the characteristic glomeruloid vascular proliferation of Fowler syndrome in all cases, but with variable extent of involvement of the central nervous system. Dystrophic calcification and necrosis were always present. Extra cranial anomalies included micrognathia (10 fetuses), cleft palate (1 fetus), cystic hygroma (2 fetuses), joint contractures (12 fetuses), and pterygia (11 fetuses). The typical proliferative vasculopathy was never observed outside the central nervous system and karyotypes were normal in the 10 fetuses studied. Fowler syndrome should be considered in the differential diagnosis of lethal multiple pterygium syndrome, fetal akinesia, and hydrocephalus in addition to classical hydranencephaly. Autopsy and study of the brain are essential to differentiate autosomal recessive Fowler syndrome from other causes of hydrocephaly and hydranencephaly, which may have a lower recurrence risk. PMID- 20014120 TI - Maximizing functional photoreceptor differentiation from adult human retinal stem cells. AB - Retinal stem cells (RSCs) are present in the ciliary margin of the adult human eye and can give rise to all retinal cell types. Here we show that modulation of retinal transcription factor gene expression in human RSCs greatly enriches photoreceptor progeny, and that strong enrichment was obtained with the combined transduction of OTX2 and CRX together with the modulation of CHX10. When these genetically modified human RSC progeny are transplanted into mouse eyes, their retinal integration and differentiation is superior to unmodified RSC progeny. Moreover, electrophysiologic and behavioral tests show that these transplanted cells promote functional recovery in transducin mutant mice. This study suggests that gene modulation in human RSCs may provide a source of photoreceptor cells for the treatment of photoreceptor disease. PMID- 20014122 TI - Morphometric analysis and classification of the facial phenotype associated with fetal alcohol syndrome in 5- and 12-year-old children. AB - Landmark-based morphometric analysis holds promise for quantitative assessment of craniofacial dysmorphology. We describe an application of facial shape analysis to characterize the facial anomalies associated with fetal alcohol syndrome (FAS) in a mixed ancestry population. Generalized Procrustes analysis, regression and discriminant function analysis were applied to stereo-photogrammetrically derived 3D coordinates of landmarks taken from 34 subjects (n = 17 FAS and n = 17 normal controls). Four shape analyses were carried out, namely a comparison of the FAS and control facial shapes at age 5, and one at age 12; a comparison of the FAS facial shapes at ages 5 and 12; and a comparison of control facial shapes at ages 5 and 12. The first two analyses showed that the FAS face is characterized by small palpebral fissures, a thin upper lip, and midfacial hypoplasia. Classification of subjects as having FAS using leave-one-out cross-validation showed that the 5-year-old group could be classified with 95.46% accuracy and the 12-year-olds with 80.13% accuracy. The third and fourth analyses revealed that the differences in facial shape between FAS individuals in different age groups were more pronounced than for control individuals, supporting the notion that FAS facial anomalies diminish with age. Geometric morphometric analysis of stereo photogrammetrically derived 3D facial landmarks allows visualization of the facial anomalies associated with FAS, as well as classification of facial shapes. PMID- 20014123 TI - Floating-Harbor syndrome associated with middle ear abnormalities. AB - Floating-Harbor syndrome is a rare syndrome of unknown etiology, which was first described in 1973. A triad of main features characterizes Floating-Harbor syndrome: short stature, characteristic face, and an expressive speech delay. We present a patient in whom the hearing thresholds improved insufficiently after placement of grommets. High-resolution CT scan of the temporal bone showed a prominent soft-tissue thickening suspected of causing fixation of the malleus, and fusion of the malleus head with the body of the incus. To our knowledge this is the first reported abnormal middle ear anatomy in a patient with Floating Harbor syndrome. A conservative treatment with hearing aids was preferred as an initial treatment in favor of a surgical exploration. PMID- 20014124 TI - Mild Wolf-Hirschhorn phenotype in a girl with unbalanced t(4p;12p) translocation without seizures. PMID- 20014125 TI - Cytodiagnosis of benign fibrous histiocytoma of rib and diagnostic dilemma: a case report. AB - Benign fibrous histiocytoma (BFH) of bone is rare in occurrence, and rib is an unusual site. There are limited case reports of this entity in the literature, and cytodiagnosis of this tumor is not described. A 24-year-old man presented with a firm mass and pain in the right lateral chest wall. Radiological investigations (plain radiograph and computed tomography) revealed a lytic bone lesion involving the 5th rib. Radiologically, giant cell tumor (GCT), BFH, and plasmacytoma were suspected. In fine-needle aspiration cytology (FNAC), admixture of benign stromal cells and scattered osteoclast type giant cells were found in the smears. Differential diagnoses of BFH, GCT (non-epiphyseal type), fibrous dysplasia, and aneurysmal bone cyst were made on cytology. Subsequent histologic examination confirmed the diagnosis of BFH. Cytologic diagnosis of BFH of rib is difficult as this tumor may mimic other giant cell containing tumors of bone in FNAC. The final diagnosis should always be made after correlation with histological, radiological, and clinical features. PMID- 20014126 TI - The process of deciding about prophylactic surgery for breast and ovarian cancer: Patient questions, uncertainties, and communication. AB - Many, women who have, or are at risk for, BRCA, 1/2 mutations or breast cancer decline prophylactic surgery, but questions remain as to how they make and experience these decisions. Our methods consisted of interviewing 32 women for 2 hr each; 19 were tested, 20 were symptomatic. Our results showed that these surgical options forced women to confront questions, involving stresses and uncertainties, regarding: implications of test results, prognoses with and without surgery, and effects of surgery (e.g., on self-image). Given these dilemmas, many women turned to doctors who then ranged in what and how they communicated about these issues, and how strongly they offered input. Some patients felt disappointed at provider non-directiveness, while others found providers too directive. Patients turned to family members and friends, who also ranged in how and what they communicated, and whether they agreed with the patient and/or each other. Many women turned to patient communities, but then had to decide how involved to be, and what information to provide or accept. These data suggest that providers and others may need to be more aware of the series of questions, involving stresses and uncertainties, that these women face, and the complex roles providers themselves may play. These data highlight needs for physicians to be able to address these issues flexibly, gauging patient preferences for information and paternalism (vs. autonomy). These data suggest areas for future research: for example, on how providers decide whether, what and how to communicate about these issues, and how these choices shape treatment decisions. PMID- 20014127 TI - Novel cardiac findings in periventricular nodular heterotopia. AB - Periventricular nodular heterotopia (PNH) is a set of neuronal migration disorders that occur during fetal development. Neurons in the brain fail to migrate from the lining of the lateral ventricles to the cortex of the brain. When the neurons fail to migrate, ectopic neuronal nodules form. Epilepsy is a common symptom of PNH. The majority of PNH cases appear to be due to mutations in filamin A, an X-linked gene. Most of the affected individuals are female because affected males typically die in utero. Filamin A anchors integral membrane proteins to the cytoskeleton by binding actin filaments in the cytoplasm. Both animal and human studies indicate that filamin A also plays a role in blood vessel development. In this report, we describe novel cardiac findings in an 18 month-old girl with PNH associated with a nonsense mutation in FLNA, including a dysplastic pulmonary valve and clefting of the mitral valve. These findings broaden the range of cardiac anomalies associated with filamin A mutations to include abnormality of the pulmonary valve and clefting of the mitral valve, consistent with a role for filamin A in valve leaflet development. PMID- 20014128 TI - Importance of intraoperative cytology in the definition of cystic solitary brain lesions. PMID- 20014129 TI - Fine-needle aspiration cytology of subcutaneous toxoplasmosis: A case report. AB - Toxoplasmosis is a common opportunistic infection in patients with AIDS in whom it typically presents as encephalitis, pneumonia, lymphadenitis, and myocarditis. Skin involvement is very rare and, to our best knowledge, Toxoplasma gondii forming a subcutaneous mass has not been reported. Here, we report the findings of an interesting case of subcutaneous toxoplasmosis with the cytological appearance of an inflammatory fibrovascular lesion in a HIV-positive patient and discuss the differential diagnosis. PMID- 20014130 TI - Steady-state level of kit ligand mRNA in goat ovaries and the role of kit ligand in preantral follicle survival and growth in vitro. AB - The aims of this study were to investigate steady-state level of Kit Ligand (KL) mRNA and its effects on in vitro survival and growth of caprine preantral follicles. RT-PCR was used to analyze caprine steady-state level of KL mRNA in primordial, primary, and secondary follicles, and in small (1-3 mm) and large (3 6 mm) antral follicles. Furthermore, ovarian fragments were cultured for 1 or 7 days in Minimal Essential Medium (MEM(+)) supplemented with KL (0, 1, 10, 50, 100, or 200 ng/ml). Noncultured (control) and cultured fragments were processed for histology and transmission electron microscopy (TEM). RT-PCR demonstrated an increase in steady-state level of KL mRNA during the transition from primary to secondary follicles. Small antral follicles had higher steady-state levels of KL mRNA in granulosa and theca cells than large follicles. After 7 days, only 50 ng/ml of KL had maintained the percentage of normal follicles similar to control. After 1 day, all KL concentrations reduced the percentage of primordial follicles and increased the percentage of growing follicles. KL at 10, 50, 100, or 200 ng/ml increased primary follicles, compared to MEM(+) after 7 days. An increase in oocyte and follicular diameter was observed at 50 ng/ml of KL. TEM confirmed ultrastructural integrity of follicles after 7 days at 50 ng/ml of KL. In conclusion, the KL mRNAs were detected in all follicular categories. Furthermore, 50 ng/ml of KL maintained the integrity of caprine preantral follicle cultured for 7 days and stimulated primordial follicle activation and follicle growth. PMID- 20014131 TI - Hypospadias in males with intrauterine growth restriction due to placental insufficiency: the placental role in the embryogenesis of male external genitalia. AB - Our aim was to define the association between early onset intra-uterine growth restriction (IUGR) due to placental insufficiency and hypospadias in males. We prospectively studied a cohort of small-for-gestational age (SGA) male infants with hypospadias managed by a multidisciplinary team over a 5-year period. Thirty SGA male infants were diagnosed with hypospadias/abnormal genitalia after birth, and four of them were diagnosed antenatally. Five cases occurred in the smaller pair of discordant IUGR twins, where the larger co-twin had normal male genitalia. Serial ultrasounds demonstrated features of early-onset IUGR in all cases at a median gestational age of 21 weeks (range 14-31weeks). Twenty-one (70%) pregnancies were subsequently complicated by absent/reversed end-diastolic flow in the umbilical arteries indicating severe IUGR, and 17 (57%) women developed severe pre-eclampsia. There were 27 (90%) live births at a median gestational age of 31 weeks (range 27-37); 23 (77%) of the neonates had birth weights <3rd centile. All newborns had normal male karyotypes. In 62% (18/29) the hypospadias was severe. A correlation was found between the severity of the IUGR and the severity of hypospadias as significantly more infants with severe hypospadias were less than the 3rd centile compared to the mild-moderate hypospadias group: 94% (17/18) versus 55% (6/11), respectively (P = 0.02). In conclusion, SGA male newborns with hypospadias exhibit a high rate of early-onset severe IUGR due to placental insufficiency. Early placental development likely influences male external genitalia formation. Careful sonographic evaluation of the genitalia is advised when early-onset placentally mediated IUGR is found. PMID- 20014132 TI - Severe cleidocranial dysplasia and hypophosphatasia in a child with microdeletion of the C-terminal region of RUNX2. AB - Cleidocranial dysplasia (CCD) is a rare autosomal dominant skeletal dysplasia due to mutations causing haploinsufficiency of RUNX2, an osteoblast transcription factor specific for bone and cartilage. The classic form of CCD is characterized by delayed closure of the fontanels, hypoplastic or aplastic clavicles and dental anomalies. Clinical reports suggest that a subset of patients with CCD have skeletal changes which mimic hypophosphatasia (HPP). Mutations in RUNX2 are detected in approximately 65% of cases of CCD, and microdeletions occur in 13%. We present clinical and radiological features in a 6-year-old child with severe CCD manifested by absence of the clavicles marked calvarial hypomineralization, osteoporosis and progressive kyphoscoliosis. HPP features included Bowdler spurs, severe osteopenia, and low alkaline phosphatase. Following negative mutation analysis of RUNX2, comparative genomic hybridization (CGH) microarray was performed. The result revealed a microdeletion in RUNX2, disrupting the C terminal part of the gene. PMID- 20014133 TI - Analysis of short-term behavioral effects of dietary cholesterol supplementation in Smith-Lemli-Opitz syndrome. AB - Smith-Lemli-Opitz syndrome (SLOS) is an inborn error of cholesterol synthesis due to mutations of 7-dehydrocholesterol reductase (DHCR7). DHCR7 catalyzes the reduction of 7-dehydrocholesterol (7DHC) to yield cholesterol in the final step of cholesterol biosynthesis. Phenotypically patients with SLOS have multiple malformations, cognitive deficits, and behavioral difficulties. Impaired DHCR7 activity results in the accumulation of 7DHC and frequently decreased cholesterol in blood and tissues. Dietary cholesterol supplementation has become standard therapy for SLOS, and anecdotal reports suggest rapid, marked clinical improvement of behavior problems. Although reported in the literature, beneficial behavioral effects of dietary cholesterol supplementation have not been formally documented through a randomized clinical trial. To address this we initiated a double-masked, placebo-controlled, cross-over trial to test the hypothesis that dietary cholesterol supplementation has rapid beneficial effects on behavior. Our primary outcome measure was the hyperactivity subscale of the Aberrant Behavior Checklist (ABC). Hyperactivity is a symptom that has been reported to respond rapidly to dietary cholesterol supplementation. Secondary outcome measures included the total ABC score and other ABC subscale scores. Ten subjects completed this study. Although the trial was done under conditions similar to those reported to induce marked behavioral changes in SLOS patients, we observed no differences between treatment and placebo phases. The results of this study call into question anecdotal reports supporting rapid behavioral benefits previously reported for dietary cholesterol supplementation in SLOS and underscore the need for a larger placebo-controlled trial. PMID- 20014135 TI - Deletion of the CUL4B gene in a boy with mental retardation, minor facial anomalies, short stature, hypogonadism, and ataxia. AB - The CUL4B gene encodes a member of Cullin-RING ubiquitin ligase complex. Point mutations in CUL4B were identified recently in patients with syndromic X-linked mental retardation (XLMR). Here, using oligoarray-based comparative genomic hybridization (array CGH), we identified a de novo deletion of the CUL4B gene in a boy with syndromic mental retardation, minor facial anomalies, short stature, delayed puberty, hypogonadism, relative macrocephaly, gait ataxia, and pes cavus, all manifestations described previously in patients with CUL4B point mutations. Interestingly, our patient also presented with aortic valvular "dysplasia" and vertebral anomalies similar to those seen in Scheuermann disease, both of which may also be part of this syndrome. This report further suggests that point mutations and deletions of the CUL4B gene lead to a recognizable phenotype. The association of facial anomalies, short stature, hypogonadism, and gait ataxia in a mentally retarded boy should prompt molecular analyses of the CUL4B gene. PMID- 20014136 TI - Two- and three-dimensional prenatal sonographic diagnosis of prune-belly syndrome. AB - We report the prenatal diagnosis of 6 cases of Prune-belly syndrome in the 2(nd) trimester. The sonographic diagnosis was based on the findings of oligohydramnios, renal anomalies, and a lower abdominal cystic mass representing the abnormal dilatation of the bladder on conventional 2-dimensional sonographic examination. We discuss the role of Doppler imaging and 3-dimensional sonography as complementary methods to conventional sonography. Four of our 6 cases were confirmed with associated defects. PMID- 20014137 TI - Contrast-enhanced sonography with abdominal virtual sonography in monitoring radiofrequency ablation of hepatocellular carcinoma. AB - BACKGROUND: Contrast-enhanced CT is regarded as the gold standard for monitoring radiofrequency ablation (RFA) of hepatocellular carcinoma (HCC). Recently, 3 dimensional volume data from CT have been used to create cross-sectional multiplanar reconstruction images. Using this technique, we can reconstruct 2 dimensional CT images identical in orientation to ultrasound (US) images, which we call virtual sonographic (VUS) images. The present prospective randomized control trial compared the number of CT scans needed to assess the efficacy of RFA of HCC using VUS-contrast-enhanced ultrasonography (CEUS) versus CT. METHOD: Subjects comprised 50 patients (50 HCCs) treated with US-guided RFA between May 2005 and August 2006, randomized to undergo assessment by CT (Group 1; 25 HCC nodules) or VUS-CEUS (Group 2; 25 HCC nodules). All patients were followed for 1 year. Primary endpoint was whether the number of CT scans could be reduced using VUS-CEUS. RESULT: Mean number of CT scans required was 1.64 +/- 0.7 in Group 1 and 1.1 +/- 0.2 in Group 2 (p < 0.001). CONCLUSION: VUS-CEUS can be used to assess the efficacy of HCC of RFA, with the potential to reduce the number of CT scans required for that purpose. PMID- 20014138 TI - Apolipoprotein E interacts with hepatitis C virus nonstructural protein 5A and determines assembly of infectious particles. AB - Chronic hepatitis C virus (HCV) infection is a major cause of liver disease worldwide. Restriction of HCV infection to human hepatocytes suggests that liver specific host factors play a role in the viral life cycle. Using a yeast-two hybrid system, we identified apolipoprotein E (apoE) as a liver-derived host factor specifically interacting with HCV nonstructural protein 5A (NS5A) but not with other viral proteins. The relevance of apoE-NS5A interaction for viral infection was confirmed by co-immunoprecipitation and co-localization studies of apoE and NS5A in an infectious HCV cell culture model system. Silencing apoE expression resulted in marked inhibition of infectious particle production without affecting viral entry and replication. Analysis of particle production in liver-derived cells with silenced apoE expression showed impairment of infectious particle assembly and release. The functional relevance of the apoE-NS5A interaction for production of viral particles was supported by loss or decrease of apoE-NS5A binding in assembly-defective viral mutants. CONCLUSION: These results suggest that recruitment of apoE by NS5A is important for viral assembly and release of infectious viral particles. These findings have important implications for understanding the HCV life cycle and the development of novel antiviral strategies targeting HCV-lipoprotein interaction. PMID- 20014139 TI - Enhancing glycoprotein sialylation by targeted gene silencing in mammalian cells. AB - Recombinant glycoproteins produced by mammalian cells represent an important category of therapeutic pharmaceuticals used in human health care. Of the numerous sugars moieties found in glycoproteins, the terminal sialic acid is considered particularly important. Sialic acid has been found to influence the solubility, thermal stability, resistance to protease attack, antigenicity, and specific activity of various glycoproteins. In mammalian cells, it is often desirable to maximize the final sialic acid content of a glycoprotein to ensure its quality and consistency as an effective pharmaceutical. In this study, CHO cells overexpressing recombinant human interferon gamma (hIFNgamma) were treated using short interfering RNA (siRNA) and short-hairpin RNA (shRNA) to reduce expression of two newly identified sialidase genes, Neu1 and Neu3. By knocking down expression of Neu3 we achieved a 98% reduction in sialidase function in CHO cells. The recombinant hIFNgamma was examined for sialic acid content that was found to be increased 33% and 26% respectively with samples from cell stationary phase and death phase as compared to control. Here, we demonstrate an effective targeted gene silencing strategy to enhance protein sialylation using RNA interference (RNAi) technology. PMID- 20014140 TI - Effect of cell lysates on retroviral transduction efficiency of cells in suspension culture. AB - Recombinant retroviruses are effective vectors able to integrate transgenes into the target cell's genome to achieve longer-term expression. This study investigates the effect of cell lysis products, a common cell culture by-product, on the transduction of suspension cells by gammaretroviral vectors. Cell lysates derived from human and murine suspension cell lines significantly increased the transduction of human TF-1 and K-562 cell lines by gibbon ape leukemia virus pseudotyped retroviral vectors without altering tropism. The transduction efficiency of TF-1 cells increased as a function of lysate concentration and decreased with increasing target cell concentrations. This was adequately predicted using a saturation equation based on the lysed-to-target cell concentration ratio, R, where: Fold increase = 1+Fold_(Max) (R/(K_(L)+R)). Lysate completely masked the effects of fibronectin when the two were added in combination. With protamine sulfate, the transduction efficiency was increased by lysate to 58% from 20% for protamine sulfate alone. Overall, the presence of cell lysate significantly influenced the outcome of the transduction process, either alone or in the presence of protamine sulfate or fibronectin. PMID- 20014141 TI - Quantification of non-specific binding of magnetic micro- and nanoparticles using cell tracking velocimetry: Implication for magnetic cell separation and detection. AB - The maturation of magnetic cell separation technology places increasing demands on magnetic cell separation performance. While a number of factors can cause sub optimal performance, one of the major challenges can be non-specific binding of magnetic nano- or microparticles to non-targeted cells. Depending on the type of separation, this non-specific binding can have a negative effect on the final purity, the recovery of the targeted cells, or both. In this work, we quantitatively demonstrate that non-specific binding of magnetic nanoparticles can impart a magnetization to cells such that these cells can be retained in a separation column and thus negatively impact the purity of the final product and the recovery of the desired cells. Through experimental data and theoretical arguments, we demonstrate that the number of MACS magnetic particles needed to impart a magnetization that is sufficient to cause non-targeted cells to be retained in the column to be on the order of 500-1,000 nanoparticles. This number of non-specifically bound particles was demonstrated experimentally with an instrument, cell tracking velocimeter, CTV, and it is demonstrated that the sensitivity of the CTV instrument for Fe atoms contained in magnetic nanoparticles on the order of 1 x 10(-15) g/mL of Fe. PMID- 20014142 TI - Micropatterning contractile C2C12 myotubes embedded in a fibrin gel. AB - Contractile C(2)C(12) myotube line patterns embedded in a fibrin gel have been developed to afford a physiologically relevant and stable bioassay system. The C(2)C(12) myotube/fibrin gel system was prepared by transferring a myotube monolayer from a glass substrate to a fibrin gel while retaining the original line patterns of myotubes. To endow the myotubes with contractile activity, a series of electrical pulses was applied through a pair of carbon electrodes placed at either side of a fibrin gel separately. The frequency and magnitude of myotube contraction were functions of the pulse frequency and duration, respectively. We found that the myotubes supported by an elastic fibrin gel maintained their line patterns and contractile activities for a longer period of time (1 week) than myotubes adhered on a conventional culture dish. PMID- 20014143 TI - Sensitive and selective detection of mycoplasma in cell culture samples using cantilever sensors. AB - In this article we report a new biosensor-based method that is more sensitive and rapid than the current approach for detecting mycoplasma in cell culture samples. Piezoelectric-excited millimeter-sized cantilever (PEMC) sensors respond to mass change via resonant frequency change. They are sensitive at femtogram level and can be used directly in liquid for label-free detection. Common cell culture contaminant, Acholeplasma laidlawii was detected in both buffer and cell culture medium. Two different sources (positive control from a commercial kit and ATCC 23206) were analyzed using antibody-immobilized PEMC sensor. Resonant frequency decrease caused by binding of A. laidlawii was monitored in real-time using an impedance analyzer. Positive detection was confirmed by a second antibody binding. The limit of detection (LOD) was lower than 10(3) CFU/mL in cell culture medium using PEMC sensor while parallel ELISA assays showed LOD as 10(7) CFU/mL. This study shows that PEMC sensor can be used for sensitive and rapid mycoplasma detection in cell culture samples. PMID- 20014144 TI - Industrial bioprocesses: beyond routine applications of established methodologies. AB - The subject matter of manuscripts by industrial authors has primarily focused on elements with perceived commercial or regulatory significance. Once published, this information interacted and ultimately influenced manuscripts from authors of other affiliations, creating the rapid advancements that culminated in the current multi-billion dollar worldwide biotechnology industry. This paper discusses trends in "solely industrial" articles published in the specific journal of Biotechnology and Bioengineering over the past five decades of this journal's lifetime. "Solely industrial" articles were defined as papers in which all the authors were affiliated with industry. Data were gathered concerning "solely industrial" article distribution and frequency, authoring companies, subject classification, and category distribution. Selected articles and their impact were related to current and past technology milestones as well as associated challenges. Suggestions for areas of greater emphasis to influence the number and subject matter of "solely industrial" articles for the journal's sixth decade were offered for consideration. PMID- 20014145 TI - Expert testimony and the effects of a biological approach, psychopathy, and juror attitudes in cases of insanity. AB - Amid growing psychological controversy and legal interest surrounding the uses of PCL-R and biological evidence in the legal system, this mock jury study assessed the effects of PCL-R and biological evidence on outcomes in an insanity defense case. A sample of 428 undergraduates read a trial transcript of an insanity defense murder case. Three variables of interest were manipulated: rebuttal illness (no mental illness, personality disorder, or psychopathy), evidentiary basis (biological or psychological), and evidentiary strength (moderately strong or moderately weak). Consistent with the hypotheses, biological evidence was more persuasive than psychological evidence, and the rebuttal was slightly more successful when the prosecution labeled the defendant as a "psychopath" than when they described him simply as "not mentally ill." PMID- 20014146 TI - Challenges in the indirect quantitation of acyl-glucuronide metabolites of a cardiovascular drug from complex biological mixtures in the absence of reference standards. AB - This paper describes the quantitation of acyl-glucuronide metabolites (M26 and M5) of a cardiovascular-drug (torcetrapib) from monkey urine, in the absence of their reference standards. LC/MS/MS assays for M1 and M4 (aglycones of M26 and M5, respectively) were characterized from normal and base-treated urine, as their respective reference standards were available. The in vivo study samples containing M26 and M5 were treated with 1 n sodium hydroxide to hydrolyze them to their respective aglycones. The study samples were assayed for M1 and M4 before and after alkaline hydrolysis and the difference in the concentrations provided an estimate of the urinary levels of M26 and M5. Prior to the main sample analysis, conditions for alkaline hydrolysis of the glucuronides were optimized by incubating pooled study samples. During incubations, a prolonged increase in M4 levels over time was observed, which is inconsistent with the base-hydrolysis of an acyl-glucuronide (expected to hydrolyze rapidly). Possible interference of the metabolite M9 (an ether-glucuronide metabolite isobaric to M4) was investigated to explain this observation using chromatographic and wet-chemistry approaches. The strategies adopted herein established that the LC/MS/MS assay and our approach were reliable. The metabolite exposure was then correlated to toxicological observations to gain initial insights into the physiological role of these metabolites. PMID- 20014147 TI - A theoretical and practical test of geographical profiling with serial vehicle theft in a U.K. context. AB - Geographical profiling is an investigative methodology sometimes employed by the police to predict the residence of an unknown offender from the locations of his/her crimes. The validity of geographical profiling, however, has not been fully explored for certain crime types. This study, therefore, presents a preliminary test of the potential for geographical profiling with a sample of 145 serial vehicle thieves from the U.K. The behavioural assumptions underlying geographical profiling (distance decay and domocentricity) are tested and a simple practical test of profiling using the spatial mean is presented. There is evidence for distance decay but not domocentricity among the spatial behaviour of car thieves from the U.K. A degree of success was achieved when applying the spatial mean on a case-by-case basis. The level of success varied, however, and neither series length in days nor number of crimes could account for the variation. The findings question previously held assumptions regarding geographical profiling and have potential theoretical and practical implications for the study and investigation of vehicle theft in the U.K. PMID- 20014149 TI - Microbiology of third and fourth branchial pouch cysts. AB - OBJECTIVES/HYPOTHESIS: To identify the most common pathogens involved in infections of third and fourth branchial pouch cysts. Third and fourth branchial pouch cyst infections are an uncommon cause of anterior neck abscesses often confused with other entities, such as thyroglossal duct cysts and thyroid abscesses leading to misdiagnosis, recurrence, and increased morbidity related to a delay in diagnosis and appropriate treatment. STUDY DESIGN: Retrospective chart and literature review. METHODS: Retrospective chart review case series of patients presenting to the Bobby R. Alford Department of Otolaryngology at Texas Children's Hospital from July 2004 to July 2008 with third and fourth branchial pouch cysts. RESULTS: A total of 11 patients were identified. All patients had left-sided lesions. Eikenella corrodens was found in 60% of cultures and was the most common organism identified in our patient group. Furthermore, 56% of the organisms isolated were anaerobic. All organisms with the exception of Staphylococcus aureus were identified as oral cavity flora. CONCLUSIONS: Third and fourth branchial pouch cysts provide a communication between the neck and the oral cavity through pyriform sinus tracts. The presence of oral cavity flora in a left anterior neck abscess should raise the suspicion of a branchial pouch anomaly, and subsequently alter therapeutic management. PMID- 20014148 TI - Genomic profiling of Richter's syndrome: recurrent lesions and differences with de novo diffuse large B-cell lymphomas. AB - Richter's syndrome (RS) represents the transformation of chronic lymphocytic leukaemia (CLL) to aggressive lymphoma and is mostly represented by diffuse large B-cell lymphoma (DLBCL), with a post-germinal centre (GC) phenotype, clonally related to the pre-existing CLL. RS has a very poor prognosis and its pathogenetic mechanisms are poorly understood. In order to gain additional hints in RS pathogenesis, we performed a genome-wide DNA profiling study of 13 RS phases and eight matched CLL phases using the Affymetrix Human Mapping 250K NspI SNP arrays. Individual genomic profiles were heterogeneous, with no individual lesions occurring in more than half of the cases. However, several observations suggest that MYC pathway might be involved in RS. The 13q13.3-qter region containing MIRHG1 (MIR-17-92), a cluster of microRNA interacting with c-MYC, was acquired at the time of transformation. The 13q gain was coupled with the gain of c-MYC and loss of TP53. Translocation of c-MYC was acquired at transformation in a fraction of cases and this event appeared mutually exclusive with gain of MIRHG1. MYCN, a c-MYC homologue, was also recurrently gained. By comparing RS with 48 de novo DLBCL, RS presented a significantly lower prevalence of deletions affecting the PRDM1 and TNFAIP3, genes on 6q, known to be associated with a post GC phenotype. In conclusion, the genomic profile of RS seems to differ from what observed in de novo DLBCL and in other transformed DLBCL. Genomic lesions occurring in RS are heterogeneous suggesting the existence of different RS subsets, possibly due to different transforming mechanisms. A deregulation of MYC pathway might represent one of the main transformation events in the pathogenesis of a subset of RS clonally related to the previous CLL. PMID- 20014151 TI - Screening for pharmaco-toxicologically relevant compounds in biosamples using high-resolution mass spectrometry: a 'metabolomic' approach to the discrimination between isomers. AB - High-resolution mass spectrometry (HRMS) enables the identification of a chemical formula of small molecules through the accurate measurement of mass and isotopic pattern. However, the identification of an unknown compound starting from the chemical formula requires additional tools: (1) a database associating chemical formulas to compound names and (2) a way to discriminate between isomers. The aim of this present study is to evaluate the ability of a novel 'metabolomic' approach to reduce the list of candidates with identical chemical formula. Urine/blood/hair samples collected from real positive cases were submitted to a screening procedure using ESI-MS-TOF (positive-ion mode) combined with either capillary electrophoresis or reversed phase liquid chromatography (LC). Detected peaks were searched against a Pharmaco/Toxicologically Relevant Compounds database (ca 50,500 compounds and phase I and phase II metabolites) consisting of a subset of PubChem compounds and a list of candidates was retrieved. Then, starting from the mass of unknown, mass shifts corresponding to pre-defined biotransformations (e.g. demethylation, glucuronidation, etc.) were calculated and corresponding mass chromatograms were extracted from the total ion current (TIC) in order to search for metabolite peaks. For each candidate, the number of different functional groups in the molecule was automatically calculated using E Dragon software (Talete srl, Milan, Italy). Then, the presence of metabolites in the TIC was matched with functional groups data in order to exclude candidates with structures not compatible with observed biotransformations (e.g. loss of methyl from a structure not bearing methyls). The procedure was tested on 108 pharmaco-toxicologically relevant compounds (PTRC) and their phase I metabolites were detected in real positive samples. The mean list length (MLL) of candidates retrieved from the database was 7.01 +/- 4.77 (median, 7; range, 1-28) before the application of the 'metabolomic' approach, and after the application it was reduced to 4.08 +/- 3.11 (median 3, range 1-17). HRMS allows a much broader screening for PTRC than other screening approaches (e.g. library search on mass spectra databases). The 'metabolomic' approach enables the reduction of the list of candidate isomers. PMID- 20014152 TI - A biplanar robot navigation system for the distal locking of intramedullary nails. AB - BACKGROUND: Distal locking is the key procedure of the intramedullary nailing operation. It is difficult to precisely locate the locking path, and intraoperative radiation exposure is also a serious problem. The advantages and disadvantages of different approaches applied in this operation are summarized. METHOD: A biplanar robot navigation system for the distal locking of intramedullary nails is presented. The novel mechanical structure and the surgical process are introduced. Then the positioning accuracy of the robot is experimented and validated. RESULTS: The positioning precision of the biplanar robot is tested, and the average error is 0.24 mm. The tibia sawbone is used for distal locking to test the accuracy of the robot navigation system. The maximal deviation between the drilling centre and the nail hole centre is 0.44 mm. CONCLUSION: The biplanar robot has the advantages of high positioning accuracy, operational stability and safety. It can be used not only for the distal locking of intramedullary nails but also for other orthopaedic surgeries that demand precise positioning. PMID- 20014153 TI - Differential effects of hydrogen peroxide and ascorbic acid on the aerobic thermosensitivity of yeast cells grown under aerobic and anoxic conditions. AB - We have previously demonstrated that in aerobically-grown cells of the yeast Saccharomyces cerevisiae, hydrogen peroxide (H(2)O(2)) increases and ascorbic acid decreases cellular thermosensitivity, as determined by the inducibility of a heat shock (HS)-reporter gene. In this work, we reveal that the aerobic thermosensitivity of anaerobically-grown yeast cells also increases in the presence of H(2)O(2), albeit differentially between cells with two different lipid profiles. In comparison to aerobically-grown fermenting cells treated with the same H(2)O(2) concentration, both these types of anaerobically-grown cells were found to be considerably less sensitive to aerobic heat shock and considerably more thermotolerant. Paradoxically, and in contrast to ascorbate pretreated aerobically-grown yeast cells, when anaerobically-grown cells were heat-shocked aerobically in the presence of the same ascorbic acid concentration, they exhibited increased thermosensitivity and decreased intrinsic thermotolerance with respect to their untreated counterparts. These findings are discussed with respect to what is currently known about the redox and physiological status of yeast cells grown aerobically and cells reoxygenated following anoxic growth. PMID- 20014154 TI - Three-dimensional A-mode ultrasound calibration and registration for robotic orthopaedic knee surgery. AB - BACKGROUND: Registration is a key step for computer-navigated robot-assisted surgery. Registration links the live patient anatomical location to the prescanned CT or MRI images, so that predesigned procedures can be performed accurately. Fiducial markers or mechanical probes are usually used to identify anatomical features or collect data points for registration. This conventional invasive approach is common; however, using ultrasound probes may provide a non invasive alternative. METHODS: This report presents investigations of selecting an A-mode ultrasound transducer, calibrating it, analysing the ultrasound signal and using it to register phantom-sawbones of tibia and femur as well as cadaveric specimens. To ensure accurate registration, the A-mode ultrasound probe is calibrated by a designed calibration system. Detailed mathematical derivation and procedures for the calibration are provided in the Appendix. The calibration and registration experiments were performed in conjunction with MAKO Surgical Corporation's Tactile Guidance System (TGS) at their headquarters and at the South Florida Spine Clinic for cadaveric experiments. RESULTS: Calibration results show that an A-mode ultrasound probe can reach the same accuracy level as a mechanical probe. By using the A-mode ultrasound probe, averaged root mean square errors (RMSE) are <0.5 mm for calibration, <1.0 mm for phantom-sawbones and <2.0 mm for cadaveric specimens. CONCLUSION: The registration results from phantom and cadaveric experiments are suitable for clinical applications. A-mode ultrasound registration is a viable option for registration of the bones in orthopaedic knee surgery but with reduced incision size. PMID- 20014155 TI - Integrating structural and epidemiological research to inform the classification of psychopathology. AB - Given the enormous influence of classification on the major clinical, research, and administrative activities of mental health professionals, understanding the true number and nature of disorders and the reasons for their comorbidity is an important public health priority. However, while studies of latent structure have yielded valuable information about disorder boundaries, their reliance on non representative samples and failure to evaluate the practical implications of structural findings has limited their ability to effect nosological change. Conversely, community epidemiology studies, which inform classification by assessing the implications of diagnostic criteria in representative samples, have been limited by their focus on mental disorders as they are currently conceptualized by the field rather than on correlates and consequences of these disorders as they actually exist in nature. I consider the potential value of integrating systematically the methods of structural research with the methods of epidemiological research, exploring five ways in which these largely independent traditions may profitably be combined to inform the next classifications of mental disorders. By capitalizing on the complementary strengths of structural and epidemiological research, an integrated approach has significant potential to advance understanding of the nature of psychopathology and improve the validity and utility of its diagnosis. PMID- 20014156 TI - Validation of a model for estimating state and local prevalence of serious mental illness. AB - This study addresses an ongoing problem in mental health needs assessment. This involves estimating the prevalence of an identified problem, specifically serious mental illness (SMI), for local areas in a reliable, valid, and cost-effective manner. The aim of the study is the application and testing of a recently introduced methodology from the field of small area estimation to determining SMI rates in the 48 contiguous US states, and in local areas of Massachusetts. It uses 'regression synthetic estimation fitted using area-level covariates', to estimate a model using data from the 2001-2002 replication of the National Comorbidity Study (n = 5593) and apply it, using 2000 STF-3C Census data, to various state and local areas in the United States. The estimates are then compared with independently collected SMI indicators. The estimates show not only face validity and internal consistency, but also predictive validity. The multiple logistic model has a sensitivity of 21.1% and a specificity of 95.1%, based largely on socio-economic disparities. Pearson r validity coefficients for the area estimates range from 0.43 to 0.75. The model generates a national estimate of SMI adults of 5.5%; for the 48 states, rates ranging from 4.7% to 7.0%; and for Massachusetts towns and cities, 1.1% to 7.5%. PMID- 20014157 TI - Genetic association between intronic variants in AS3MT and arsenic methylation efficiency is focused on a large linkage disequilibrium cluster in chromosome 10. AB - Differences in arsenic metabolism are known to play a role in individual variability in arsenic-induced disease susceptibility. Genetic variants in genes relevant to arsenic metabolism are considered to be partially responsible for the variation in arsenic metabolism. Specifically, variants in arsenic (3+ oxidation state) methyltransferase (AS3MT), the key gene in the metabolism of arsenic, have been associated with increased arsenic methylation efficiency. Of particular interest is the fact that different studies have reported that several of the AS3MT single nucleotide polymorphisms (SNPs) are in strong linkage-disequilibrium (LD), which also extends to a nearby gene, CYP17A1. In an effort to characterize the extent of the region in LD, we genotyped 46 SNPs in a 347,000 base region of chromosome 10 that included AS3MT in arsenic-exposed subjects from Mexico. Pairwise LD analysis showed strong LD for these polymorphisms, represented by a mean r(2) of 0.82, spanning a region that includes five genes. Genetic association analysis with arsenic metabolism confirmed the previously observed association between AS3MT variants, including this large cluster of linked polymorphisms, and arsenic methylation efficiency. The existence of a large genomic region sharing strong LD with polymorphisms associated with arsenic metabolism presents a predicament because the observed phenotype cannot be unequivocally assigned to a single SNP or even a single gene. The results reported here should be carefully considered for future genomic association studies involving AS3MT and arsenic metabolism. PMID- 20014158 TI - Efficacy of vaporised ethyl formate/carbon dioxide formulation against stored grain insects: effect of fumigant concentration, exposure time and two grain temperatures. AB - BACKGROUND: The ethyl formate/carbon dioxide (CO(2)) formulation Vapormate is a rapid-acting fumigant being developed for the control of stored-grain insects. The effects have been investigated of concentration, exposure times of 1, 3, 24 and 72 h and two grain temperatures, 15 and 25 degrees C, on its efficacy against mixed-stage cultures of Sitophilus oryzae (L.) Tribolium castaneum (Herbst) and strongly phosphine-resistant Rhyzopertha dominica (F.) strain QRD569. RESULTS: High mortalities (> or = 92%) of mixed-stage cultures of all three species were obtained when grain was fumigated with the formulation (193 g m(-3) ethyl formate) for 1 h. Complete control of R. dominica QRD569 and T. castaneum was achieved with 63 and 76 g m(-3) ethyl formate respectively, with exposure for 24 h, whereas mean mortality of S. oryzae was 86% under the same conditions. Mortalities of S. oryzae juvenile stages were significantly lower than adults under the conditions tested, which was due to pronounced tolerance of mid-stage pupae to the fumigant. Reducing grain temperature from 25 to 15 degrees C had no effect on insect mortality. CONCLUSION: Ethyl formate/CO(2) formulation is highly effective against stored-grain insects over a range of concentrations and exposure times. Efficacious fumigations were conducted in as little as 1 h, and a strongly phosphine-resistant R. dominica strain was readily controlled with the fumigant. PMID- 20014159 TI - Review of the literature on the effects of caring for a patient with cancer. AB - OBJECTIVE: To adequately help family caregivers (FCs) of cancer patients, clinicians need to understand the complexity of the problems and responsibilities associated with cancer patients illness that FCs experience. METHODS: This systematic review identified the types of problems and burdens that FCs of cancer patients experience during the patient's illness. We also analyzed the language caregivers use to communicate their problems and responsibilities related to caregiving for the cancer patient. RESULTS: Of 2845 titles identified, 192 articles met the inclusion criteria and are included in this review. Of these, 164 were research-based. In addition to FC responsibilities and the impact of being a caregiver on daily life, a number of other physical, social, and emotional problems related to caregiving for these FCs were identified. CONCLUSION: A substantial evidence base supports the conclusion that FCs experience many difficult problems and increased responsibilities during and after the patient is undergoing treatment and rehabilitation for cancer. The insights gained from this review will help researchers and clinicians to understand the complexity of problems and responsibilities FCs experience. This understanding may encourage them to include support for FCs as part of total or holistic patient care. However, more research is needed to better understand the variations in caregiving experiences over time; how the caregiving perspective is influenced by different cultural, ethnic, or socioeconomic backgrounds as well as gender and age; and how problems and responsibilities related to caregiving interfere with daily life. PMID- 20014160 TI - Social support buffers the impact of functional impairments on caregiver psychological well-being in the context of brain tumor and other cancers. AB - OBJECTIVE: This study investigated the association between functional impairments of individuals with cancer and caregiver psychological well-being, and examined the moderating effect of social support. METHODS: Sixty-three caregivers (71% female) of individuals with brain tumor (n=27) and other cancers (n=36) were recruited from community services. Caregivers rated their psychological well being on the World Health Organisation Quality of Life measure Brief version, social support on a brief version of the Social Support Questionnaire, and the individuals' functional impairments on the Patient Competency Rating Scale. RESULTS: For caregivers of individuals with brain tumor, better psychological well-being was associated with lower functional impairment in all domains (r(s)=0.33-38, p<0.05), except for cognitive difficulties. For caregivers of individuals with other cancers, better psychological well-being was associated with lower functional impairment in all domains (r(s)=0.30-0.49, p<0.05), with the exception of activities of daily living. For the total caregiver sample, better psychological well-being was significantly correlated with overall functional impairment (r=0.34, p<0.005) and satisfaction with support (r=0.40, p<0.005). Caregivers supporting individuals with greater functional impairment had better psychological well-being if they were highly satisfied with their social support. CONCLUSIONS: Effective social support is particularly important for caregivers who support individuals with poorer functional status, and this study highlights the need to evaluate caregiver social support interventions in the context of cancer. PMID- 20014161 TI - Characterization of an exception to the 'even-electron rule' upon low-energy collision induced decomposition in negative ion electrospray tandem mass spectrometry. AB - Electrospray-generated precursor ions usually follow the 'even-electron rule' and yield 'closed shell' fragment ions. We characterize an exception to the 'even electron rule.' In negative ion electrospray mass spectrometry (ES-MS), 2 (ethoxymethoxy)-3-hydroxyphenol (2-hydroxyl protected pyrogallol) easily formed a deprotonated molecular ion (M-H)(-) at m/z 183. Upon low-energy collision induced decomposition (CID), the m/z 183 precursor yielded a radical ion at m/z 124 as the base peak. The radical anion at m/z 124 was still the major fragment at all tested collision energies between 0 and 50 eV (E(lab)). Supported by computational studies, the appearance of the radical anion at m/z 124 as the major product ion can be attributed to the combination of a low reverse activation barrier and resonance stabilization of the product ions. Furthermore, our data lead to the proposal of a novel alternative radical formation pathway in the protection group removal of pyrogallol. PMID- 20014162 TI - A panel study to evaluate quality of life assessments in patients suffering from allergic rhinitis after treatment with a Chinese herbal nasal drop. AB - Allergic rhinitis impairs quality of life (QOL). To assess the changes in QOL of patients with perennial allergic rhinitis (PAR) after treatment with Allergic Rhinitis Nose Drops (ARND), 35 patients were divided into 2 groups in a randomized, double-blinded and placebo-controlled study, with a cross-over arrangement over 7 weeks, applying ARND or placebo. Group A (n = 20) started with ARND first for 2 weeks followed by a 3-week washout before placebo for the last 2 weeks, while Group B (n = 15) started with placebo first and finished with ARND after washout. The changes in Clinical Symptoms Score (CSS) and QOL were observed. A decrease in CSS was observed in patients of both groups after treatment with ARND, but no change was observed with the placebo. Group A patients also showed significant improvements in complexion and sleep (P < 0.05 for both) after treatment with ARND, but no change with the placebo. Group B patients showed significant improvements in appetite and digestion (P < 0.01) as well as joy (P < 0.05) after cross-over treatment with ARND, but no change with the placebo. ARND may have a therapeutic effect by relieving clinical symptoms and improving the QOL in patients with PAR. PMID- 20014163 TI - In vitro cytotoxic, antiprotozoal and antimicrobial activities of medicinal plants from Vanuatu. AB - Sixty-three extracts obtained from 18 plants traditionally used in the South Pacific archipelago Vanuatu for the treatment of infectious diseases were screened for antimicrobial and antiprotozoal activities. In addition, the extracts were subjected to a detailed analysis on cytotoxic effects toward a panel of human cancer cell lines, designed as a smaller version of the NCI60 screen. Intriguingly, 15 plant extracts exhibited strong cytotoxic effects specific for only one cancer cell line. Extracts of the leaves of Acalypha grandis Benth. significantly affected Plasmodium falciparum without showing obvious effects against the other protozoa tested. The leaves of Gyrocarpus americanus Jacq. displayed significant activity against Trypanosoma b. brucei and the leaves of Tabernaemontana pandacaqui Lam. I as well as the stems of Macropiper latifolium (L.f.) against Trypanosoma cruzi. In contrast none of the extracts showed relevant antibacterial or antifungal activity. PMID- 20014164 TI - A proposed model for dragline spider silk self-assembly: insights from the effect of the repetitive domain size on fiber properties. AB - Dragline spider silk has been intensively studied for its superior qualities as a biomaterial. In previous studies, we made use of the baculovirus mediated expression system for the production of a recombinant Araneus diadematus spider silk dragline ADF4 protein and its self-assembly into intricate fibers in host insect cells. In this study, our aim was to explore the function of the major repetitive domain of the dragline spider silk. Thus, we generated an array of synthetic proteins, each containing a different number of identical repeats up to the largest recombinantly expressed spider silk to date. Study of the self assembly properties of these proteins showed that depending on the increasing number of repeats they give rise to different assembly phenotypes, from a fully soluble protein to bona fide fibers with superior qualities. The different assembly forms, the corresponding chemical resistance properties obtained as well as ultrastructural studies, revealed novel insights concerning the structure and intermolecular interactions of the repetitive and nonrepetitive domains. Based on these observations and current knowledge in the field, we hereby present a comprehensive hypothetical model for the mechanism of dragline silk self-assembly and fiber formation. PMID- 20014165 TI - Complications of low-dose, echo-guided alcohol septal ablation. AB - BACKGROUND: Alcohol septal ablation (ASA) is a catheter-based intervention that has been used as an alternative to surgical myectomy in highly symptomatic patients with hypertrophic obstructive cardiomyopathy (HOCM). METHODS: This retrospective study was designed to evaluate the incidence of major complications in the mid-term follow-up of low-dose (1-2.5 ml of ethanol), echo-guided alcohol septal ablation. RESULTS: A total of 101 consecutive patients (56 +/- 15 years) with highly symptomatic HOCM were enrolled. At 6 months, there was a significant decrease in resting outflow gradient accompanied by reduction in basal septal diameter and improvement in symptoms (P < 0.01). Two patients (2%) experienced procedural ventricular tachycardias terminated by electrical cardioversion. A total of 87 patients (86%) underwent an uneventful postprocedural hospital stay. The postprocedural complete heart block occurred in 10 patients (10%), and subsequent permanent pacemaker was implanted in four cases (4%). Sustained ventricular arrhythmias requiring electrical cardioversion occurred in four patients (4%) within postprocedural hospital stay. Subsequently, ICD was not implanted in any of these cases. The patients were repeatedly examined by Holter ECG monitoring, and in the mid-term follow-up (6-50 months), they stayed asymptomatic and without any ventricular arrhythmias. CONCLUSION: This study demonstrates the same early incidence of complete heart block requiring permanent pacemaker implantation (4%) and sustained ventricular arrhythmias following low dose, echo-guided ASA. PMID- 20014166 TI - Usefulness of prescription monitoring programs for surveillance--analysis of Schedule II opioid prescription data in Massachusetts, 1996-2006. AB - PURPOSE: Electronic prescription monitoring programs (PMPs) have been developed in many states as a public health surveillance tool. We analyze herein 11 years of Massachusetts PMP data to evaluate trends in opioid prescribing, dispensing, and usage. METHODS: Prescription records from the Massachusetts PMP for Schedule II opioids from fiscal year 1996 to 2006 were analyzed. 'Questionable activity' (potential 'doctor shopping') estimates were based on individual use of multiple prescribers and pharmacies, and early refills. RESULTS: The number of prescriptions, doses prescribed, and individuals receiving Schedule II prescription opioids steadily increased from 1996 to 2006. Most individuals (87.5%) used 1-2 prescribers, 1-2 pharmacies, and had no early refills (2006). The greater the number of prescribers used, the greater the number of pharmacies used. When defined as the use of >or=4 prescribers and >or=4 pharmacies, questionable activity accounted for 2748 individuals, 47 953 prescriptions, and 2 966 056 doses (2006). The Schedule II opioid most highly associated with questionable activity was short-acting oxycodone. CONCLUSIONS: PMPs can become a useful public health surveillance tool to monitor the medical and non-medical use of prescription opioids and to inform public health and safety policy. PMID- 20014167 TI - Measuring exacerbations in obstructive lung disease. AB - PURPOSE: Using hospitalization always has been seen as a solid measurement for exacerbation in pharmacoepidemiology, but might lead to an underestimation of disease exacerbation because of a trend towards outpatient care. The aim of this study was to quantify the incidence of different exacerbation markers in obstructive lung disease and to identify predictors for these exacerbation markers. METHODS: We conducted a cohort study using the PHARMO record linkage system, including demographic details and complete medication histories of more than two million community-dwelling residents in the Netherlands from 1985 onwards. Eligible patients were adult users of inhaled corticosteroids (ICS). Outcome parameters were hospitalization and short courses of systemic corticosteroids. Patients were allowed to have multiple exacerbations during follow-up. RESULTS: We identified 5327 patients. During follow-up, 8635 exacerbations occurred in 2332 patients with a trend in time towards treating exacerbations out of the hospital (p-value 0.003). Of all patients with exacerbations, 73% was not hospitalized during follow-up. Exacerbations were associated with high-dose ICS use (adjusted RR 1.4; 95% CI 1.2-1.7) and chronic systemic corticosteroid use (adjusted RR 1.9; 95%CI 1.6-2.2). CONCLUSIONS: Using hospitalization only as exacerbation marker leads to underestimating the exacerbation rate, because of exacerbation treatment out of the hospital. Patients with obstructive lung disease using chronic systemic corticosteroids or high-dose ICS use are more prone to exacerbations. This implies that these patients should be monitored carefully to prevent recurrent exacerbations which are detrimental for their prognosis and quality of life. PMID- 20014168 TI - PAPP-A and free ss-hCG stability in first trimester serum using PerkinElmer AutoDELFIA and DELFIA Xpress systems. AB - BACKGROUND: In this study we aim to investigate the stability of free-beta-hCG and PAPP-A over time in serum and whole blood in typical routine temperatures. METHODS: Serum pools were stored under the following temperatures: 30 degrees C, room temperature, refrigerator temperature and -20 degrees C, for up to 240 days. Stability of the markers in whole blood was examined in a shorter study and compared to serum. Samples were analysed using the AutoDELFIA and DELFIA Xpress analysers. RESULTS: On the AutoDELFIA, considering a 10% change acceptable, PAPP A levels are stable in serum for 142 days at refrigerator temperature, 37 days at room temperature and 20 days at 30 degrees C. Free-beta hCG is stable in serum for 94 days at refrigerator temperature, 3 days at room temperature and 12 h at 30 degrees C. There was no significant change with either analyte after -20 degrees C storage for up to 240 days or after six repeated freeze-thaw cycles. In whole blood, free-beta hCG levels increased more rapidly compared to serum, especially at 30 degrees C. CONCLUSION: Normal handling of samples is only likely to minimally effect the risk assessment of chromosomal anomalies. However, careful attention should be paid to minimise the increase of free-beta hCG levels in samples shipped as whole blood. PMID- 20014169 TI - Second trimester prenatal diagnosis of rhizomelic chondrodysplasia punctata type 1 on ultrasound findings. PMID- 20014170 TI - Signal detection for vaccine side effects that have not been specified in advance. AB - An information-theoretic definition of similarity proposed by Dekang Lin in 1998 appears to have attractive properties for identifying medically similar cases in a large database. Examination of similar cases that appear following vaccination may provide a way of detecting previously unexpected associations. A key element here is the inclusion of shared time-since-vaccination as a dimension of similarity while considering concordance on many other features. The system is potentially much more sensitive to patterns with multiple features than are very large, high-dimension contingency tables. PMID- 20014172 TI - Massive pulmonary embolism and paradoxical migration during surgical embolectomy: role of transesophageal echocardiogram. AB - Patent foramen ovale (PFO) in the setting of venous thromboembolism is associated with paradoxical embolization. We describe a patient who presented with pulmonary embolism, underwent pulmonary embolectomy, and postoperatively developed paradoxical embolization to the lower extremity.A 27-year-old African American male presented to the hospital with shortness of breath and midsternal chest pain along with neck vein distention. A CT scan with contrast showed the presence of a saddle embolus in both pulmonary arteries. The next day, the patient developed right ventricular failure and hypotension. The patient was taken to the operating room for a pulmonary embolectomy. Postoperatively, the patient developed acute left lower extremity ischemia. The origin of the embolus was suspected to be cardiac. A transesophageal echocardiogram (TEE) revealed thrombus on the mitral valve and a PFO with right to left shunt. At this point vascular surgery for revascularization of the left lower extremity was performed. Two days later, the patient was taken for a repeat cardiac surgery and the left-sided thrombus was removed along with a closure of the PFO.This case signifies the importance of complete TEE and a search for PFO in patients with massive pulmonary embolism especially prior to surgical embolectomy because hemodynamic disturbances of pulmonary embolism and surgical embolectomy may cause migration of the thrombus from the right side to the left side of the heart. PMID- 20014173 TI - Growth differentiation factor 15 and coronary collateral formation. AB - BACKGROUND: The coronary collateral circulation can reduce sudden cardiac death,myocardial cell loss,and infarct size.Growth differentiation factor 15(GDF 15),a member of the transforming growth factor-beta (TGF-beta) superfamily,has been reported to have a prognostic predicting value in coronary artery disease. HYPOTHESIS: GDF-15 can be related with the extent of collateral formation. OBJECTIVE: Growth differentiation factor 15 (GDF-15), a member of the transforming growth factor-beta (TGF-beta) superfamily, has been reported to have a prognostic predicting value in coronary artery disease. We sought to investigate whether GDF-15 is related to coronary collateral development in patients with coronary heart disease. METHODS: A cross-sectional study was performed in 201 patients, who were admitted for selective coronary angiography. Patients were divided into 3 groups based on Rentrop's classification of coronary collaterals. Group 1: patients with coronary collateral presence, which was defined by Rentrop's grade 1-3 collateral development. Group 2: patients with grade 0 collateral development. Group 3: control group were patients with a normal coronary angiogram. The levels of plasma GDF-15, asymmetric dimethylarginine (ADMA), and soluble Fms-related tyrosine kinase-1 (sFLT-1) were compared among the 3 groups. RESULTS: There were significant statistical differences in plasma sFLT-1, ADMA, and GDF-15 concentrations among the different collateral groups. The correlations between Rentrop's grade and the cytokines were significant. A positive correlation was found between Rentrop's grade and GDF-15 (r = 0.187, P < 0.05). The correlations between the levels of plasma sFLT 1, ADMA, and Rentrop's grade were significant, with the correlation coefficient of r = 0.181, P < 0.05 (sFLT-1) and r = - 0.646, P < 0.001 (ADMA), respectively. CONCLUSIONS: Our findings suggest that GDF-15 levels increase with the extent of collateral formation. In that case, the patients with a higher level of GDF-15 may predict more severe coronary stenosis, which has a higher probability to develop collaterals. PMID- 20014174 TI - Trends in the non-steroidal anti-inflammatory drug market in six Central-Eastern European countries based on retail information. AB - PURPOSE: The objective of this study was to evaluate the non-steroidal anti inflammatory drug market of six Central and Eastern European countries. Trends and similarities were compared across the examined countries. METHODS: The Intercontinental Marketing Service Health database was used to determine consumption data between the years 2000 and 2007. We applied the anatomical therapeutical chemical-defined daily dose method, focussing on three major non steroidal anti-inflammatory drug groups: conventional non-steroidal anti inflammatory drugs, 'stronger cyclooxygenase 2 inhibitors' (all together as: non cyclooxygenase 2 selective non-steroidal anti-inflammatory drugs) and selective cyclooxygenase 2 inhibitors. The main outcome measure was defined daily dose/1000 inhabitants/day. Different active agents have been distinguished between the three major groups. RESULTS: In total the non-steroidal anti-inflammatory drug group reached a 42.82-74.17 defined daily dose/1000 inhabitants/day volume in 2007, with an average total increase of 25.1% between 2002 and 2007. In the conventional non-steroidal anti-inflammatory drug group, diclofenac and ibuprofen have attained the highest consumption. Our results show a notable increase (325%, 2002-2007) of the 'stronger cyclooxygenase 2 inhibitor group' (nimesulide and meloxicam). Trends of selective cyclooxygenase 2 inhibitor volumes differ within the observed countries. CONCLUSION: Differences in the six countries concerning their NSAID consumption and market trends could not be explained with the inequalities in patient characteristics. The conventional NSAID retail gave the majority of the total NSAID market. The consumption of selective COX2 inhibitors in all of the six countries were much lower than in the US or Australia. The NSAID risk profile in the region is comparable to previous studies in other countries. PMID- 20014175 TI - Acute coronary syndromes: clinical characteristics, management, and outcomes at the American University of Beirut Medical Center, 2002-2005. AB - OBJECTIVES: Data on acute coronary syndromes (ACS) in developing countries is scarce. In this report, we analyze the temporal trends in the management and outcomes of a large series of ACS patients hospitalized at the American University of Beirut Medical Center (AUBMC), a tertiary referral university hospital located in a middle income Middle Eastern developing country. METHODS: A total of 1025 consecutive patients hospitalized and discharged with the diagnosis of ACS were enrolled between 2002 and 2005. The utilization of evidence-based therapies and in-hospital outcomes were determined. RESULTS: The study enrolled 228 patients (22%) with ST-elevation myocardial infarction (STEMI), 275 patients (27%) with non-ST-elevation myocardial infarction (NSTEMI), and 522 patients (51%) with unstable angina. The STEMI group was younger and had a higher percentage of men. The utilization rates of coronary angiography and percutaneous coronary intervention (PCI) were highest in the STEMI group. Comparison to earlier ACS data (1997-1998) from the same hospital, showed an increase in the utilization of reperfusion therapy, coronary angioplasty, bypass surgery, aspirin, beta-blockers, angiotensin-converting enzymes (ACE), angiotensin receptor blockers (ARB), and statins over the past decade (P < .05). This was associated with a significant decrease in hospital mortality (13%-7.7%, P < .01). CONCLUSIONS: This study analyzes one of the largest series of ACS patients reported from a single center in a developing country. The utilization of evidence-based therapies in the management of ACS at AUBMC has improved significantly over the past decade with an associated decrease in hospital mortality. PMID- 20014176 TI - Retracted: Relation of atherosclerotic cardiovascular events to progression of aortic stenosis in older men. PMID- 20014177 TI - A remnant sewing needle in the right ventricle as a cause of chest pain. AB - Sewing needles, albeit a rare case of penetrating cardiac injury, are potentially life-threatening. We report a case of a self-inflicted remnant intracardiac sewing needle which is diagnosed with nonspecific chest pain and successfully extracted 23 months after its insertion. PMID- 20014178 TI - Muscle toxicity with statins. AB - PURPOSE: Statins rarely cause serious muscle toxicity and rhabdomyolysis. The aim of our investigation was to identify and quantify potential risk factors for statin-induced rhabdomyolysis. METHODS: All cases of suspected adverse reactions to statins reported to the Swedish Adverse Drug Reactions Advisory Committee until 15 September 2006 containing the codes myalgia, myopathy, increased serum creatine kinase (CK), myoglobinuria or rhabdomyolysis were included in the study. Cases were classified into different CK categories, where cases with CK levels >10 times the upper limit of normal (ULN) laboratory range were compared with cases with normal CK levels (in some analyses cases with CK not measured were also included as controls). Fisher's test and multiple logistic regression were used to test the degree of association. RESULTS: A total of 338 cases with muscle toxicity were identified. CK had not been measured in 148 cases. Of the remaining 190 cases, 59 were classified as rhabdomyolysis, 62 had CK increases below the level of rhabdomyolysis, 69 had normal CK and 2 contained insufficient information to classify the degree of CK increase. A high statin dose and concomitant interacting drug treatment were over-represented among cases with rhabdomyolysis compared with cases with normal CK. Renal disease and unusual strenuous muscular activity were also associated with an increased risk of rhabdomyolysis when the control group included cases with CK not measured. CONCLUSION: Results from our study support previous studies indicating that the risk of rhabdomyolysis with statin treatment increases with increase in systemic exposure to the statin. Renal disease and unusual strenuous muscular activity may also contribute to an increased risk of rhabdomyolysis. PMID- 20014179 TI - Water-related occupations and diet in two Roman coastal communities (Italy, first to third century AD): correlation between stable carbon and nitrogen isotope values and auricular exostosis prevalence. AB - The reconstruction of dietary patterns in the two Roman imperial age coastal communities of Portus and Velia (I-III AD) by means of stable isotope analysis of bone remains has exposed a certain degree of heterogeneity between and within the two samples. Results do not correlate with any discernible mortuary practices at either site, which might have pointed to differential social status. The present study tests the hypothesis of a possible connection between dietary habits and occupational activities in the two communities. Among skeletal markers of occupation, external auricular exostosis (EAE) has proved to be very informative. Clinical and retrospective epidemiological surveys have revealed a strong positive correlation between EAE development and habitual exposure to cold water. In this study, we show that there is a high rate of occurrence of EAE among adult males in both skeletal samples (21.1% in Portus and 35.3% in Velia). Further, there is a statistically significant higher prevalence of EAE among those individuals at Velia with very high nitrogen isotopic values. This points to fishing (coastal, low-water fishing) as the sea-related occupation most responsible for the onset of the ear pathology. For Portus, where the consumption of foods from sea and river seems to be more widespread through the population, and where the scenario of seaport and fluvial activities was much more complex than in Velia, a close correlation between EAE and fish consumption by fishermen is less easy to establish. PMID- 20014180 TI - Patient focused drug surveillance of elderly patients in nursing homes. AB - PURPOSE: To examine whether patient focused drug surveillance was associated with a higher quality of drug treatment at nursing homes. METHODS: An intervention study in four nursing homes in Sweden and four other nursing homes served as controls. At the intervention nursing homes physicians focused on patients' health status as a baseline for further ongoing medication. The outcomes were mortality, health care consumption, and number of drugs, health status and evaluations as parameters for "quality of drug treatment". RESULTS: There were no significant differences in mortality rates. Health care consumption and polypharmacy were extensive in both groups. There was a significant reduction of number of drugs used per patient at the intervention homes during the study (p < 0.05). Monitoring and evaluation of the effects of medications were significantly more frequent at the intervention homes (p < 0.01). CONCLUSIONS: The intervention resulted in significant positive results in relation to "quality of drug treatment", a shift in health care utilization with concomitant use of fewer drugs. The study showed an extreme shortage of monitoring of health status and surveillance of the effects of drugs in the elderly. PMID- 20014181 TI - Racial disparities in the use of radiotherapy after breast-conserving surgery: a national Medicare study. AB - BACKGROUND: In prior studies, the use of standard breast cancer treatments has varied by race, but previous analyses were not nationally representative. Therefore, in a comprehensive, national cohort of Medicare patients, racial disparities in the use of radiotherapy (RT) after breast-conserving surgery (BCS) for invasive breast cancer were quantified. METHODS: A national Medicare database was used to identify all beneficiaries (age >65 years) treated with BCS for incident invasive breast cancer in 2003. Claims codes identified RT use, and Medicare demographic data indicated race. Logistic regression modeled RT use in white, black, and other-race patients, adjusted for demographic, clinical, and socioeconomic covariates. RESULTS: Of 34,080 women, 91% were white, 6% were black, and 3% were another race. The mean age of the patients was 76 +/- 7 years. Approximately 74% of whites, 65% of blacks, and 66% of other-race patients received RT (P < .001). After covariate adjustment, whites were found to be significantly more likely to receive RT than blacks (odds ratio, 1.48; 95% confidence interval, 1.34-1.63 [P < .001]). Disparities between white and black patients varied by geographic region, with blacks in areas of the northeastern and southern United States demonstrating the lowest rates of RT use (57% in these regions). In patients age <70 years, racial disparities persisted. Specifically, 83% of whites, 73% of blacks, and 78% of other races in this younger group received RT (P < .001). CONCLUSIONS: In this comprehensive national sample of older breast cancer patients, substantial racial disparities were identified in RT use after BCS across much of the United States. Efforts to improve breast cancer care require overcoming these disparities, which exist on a national scale. PMID- 20014182 TI - Neandertal mandibles from the Sima de las Palomas del Cabezo Gordo, Murcia, southeastern Spain. AB - The Middle Paleolithic levels of the Sima de las Palomas have yielded eight partial mandibles (Palomas 1, 6, 7, 23, 49, 59, 80, and 88). Palomas 7, 49, 80, and 88 are immature, and Palomas 49, 59, 80, and 88 are among the latest Neandertals (approximately 40,000 cal BP). Palomas 1 is geologically older (approximately 50,000-60,000 cal BP), and the other three were found ex situ. The mandibles exhibit a suite of characteristics that align them with the Neandertals among later Pleistocene humans, including symphyseal morphology, symphyseal orientation, corpus robusticity, distal mental foramen position, retromolar space presence, wide immature dental arcade, and high-coronoid process with an asymmetrical mandibular notch. However, Palomas 6 lacks a retromolar space, Palomas 59 has a narrow lateral corpus, and Palomas 80 has a mesial mental foramen and open mandibular foramen. The Palomas mandibles therefore help to document that the late Middle Paleolithic of southern Iberia was the product of Neandertals. They also reinforce the presence of variability in both metric and discrete aspects of Neandertal mandibular morphology, both within and across samples, some of which may be temporal and/or geographic in nature. PMID- 20014184 TI - Cancer-related concerns of spouses of women with breast cancer. AB - OBJECTIVE: To describe spouses' reported cancer-related demands attributed to their wife's breast cancer, and to test the construct and predictive validity of a brief standardized measure of these demands. METHODS: Cross-sectional and longitudinal data were obtained from 151 spouses of women newly diagnosed with non-metastatic breast cancer. Descriptive statistics were computed to describe spouses' dominant cancer-related demands, and multivariate regression analyses tested the construct and predictive validity of the standardized measure. RESULTS: Five categories of spouses' cancer-related demands were identified, such as concerns about spouses' own functioning, wife's well-being and response to treatment, couples' sexual activities, the family's and children's well-being, and the spouses' role in supporting their wives. A 33-item short version of the standardized measure of cancer demands demonstrated construct and predictive validity that was comparable to a 123-item version of the same questionnaire. Greater numbers of illness demands occurred when spouses were more depressed and had less confidence in their ability to manage the impact of the cancer (F=18.08 (3,103), p<0.001). Predictive validity was established by the short form's ability to significantly predict the quality of marital communication and spouses' self-efficacy at a 2-month interval. CONCLUSION: The short version of the standardized measure of cancer-related demands shows promise for future application in clinic settings. Additional testing of the questionnaire is warranted. Spouses' breast cancer-related demands deserve attention by providers. In the absence of assisting them, spouses' illness pressures have deleterious consequences for the quality of marital communication and spouses' self confidence. PMID- 20014185 TI - Multidetector computed tomography in an adult patient with silent patent ductus arteriosus. PMID- 20014183 TI - A randomized, controlled, double-blind, pilot study of milk thistle for the treatment of hepatotoxicity in childhood acute lymphoblastic leukemia (ALL). AB - BACKGROUND: Despite limited preclinical and clinical investigations, milk thistle (MT) is often used for the treatment of chemotherapy-associated hepatotoxicity. Limited treatment options exist for chemotherapy-related hepatoxicity. Given the wide use of MT, the authors investigated MT in both the laboratory and a clinical setting. METHODS: In a double-blind study, children with acute lymphoblastic leukemia (ALL) and hepatic toxicity were randomized to MT or placebo orally for 28 days. Liver function tests were evaluated during the study period. To assess MT in vitro, the authors evaluated supratherapeutic concentrations in an ALL cell line. RESULTS: Fifty children were enrolled. No significant differences in frequency of side effects, incidence and severity of toxicities, or infections were observed between groups. There were no significant changes in mean amino alanine transferase (ALT), aspartate amino transferase (AST), or total bilirubin (TB) at Day 28. At Day 56, the MT group had a significantly lower AST (P = .05) and a trend toward a significantly lower ALT (P = .07). Although not significantly different, chemotherapy doses were reduced in 61% of the MT group compared with 72% of the placebo group. In vitro experiments revealed no antagonistic interactions between MT and vincristine or L-asparaginase in CCRF CEM cells. A modest synergistic effect with vincristine was observed. CONCLUSIONS: In children with ALL and liver toxicity, MT was associated with a trend toward significant reductions in liver toxicity. MT did not antagonize the effects of chemotherapy agents used for the treatment of ALL. Future study is needed to determine the most effective dose and duration of MT and its effect on hepatotoxicity and leukemia-free survival. PMID- 20014187 TI - Three year follow-up of the sirolimus-eluting stent and the paclitaxel-eluting stent in daily practice. AB - BACKGROUND: Although the efficacy and safety of Cypher and Taxus stents for particular patients and lesions had been proven by random clinical trials, their long-term comparative effect for unselected Chinese patients in daily practice is still unknown. METHODS: From April 2004 to April 2005, 682 consecutive patients who were implanted with a Cypher stent (C group: n = 456, 57.3 +/- 10.8 years old) or a Taxus stent (T group: n = 226, 57.2 +/- 10.2 years old) in our center were analyzed. A mean 3 year clinical and 7 month angiographic follow-up was performed. RESULTS: There was no difference in major adverse cardiac events (MACE) rates in the C and T groups (16.4% vs 21.7%; P = .113), and there was also no difference in cardiac death or myocardial infarction (MI) rates in both groups (2.6% vs 1.8%; P = .598, 3.3% vs 3.5%; P = .826 respectively), but target vessel revascularization (TVR) was higher in the T group (10.5% vs 16.4%; P = .036). There was no difference in total stent thrombosis or its components in both groups (total: 3.9% vs 3.5%; P = 1.000, early: 0.2% vs 0.9%; P = .256, late: 0.7% vs 1.3%; P = .404, and very late: 3.1% vs 1.3%; P = .201 respectively). A 7 month angiographic follow-up indicated that both in-stent and in-segment restenosis rate were significantly lower in the C group (11.1% vs 21.0%; P = .034 and 12.2% vs 24.0%; P = .018), and in-stent and in-segment late loss were significantly smaller in the C group (0.18 +/- 0.08 mm vs 0.53 +/- 0.38 mm; P = .000 and 0.17 +/- 0.09 mm vs 0.38 +/- 0.19 mm; P = .000). CONCLUSION: Results from this 3 year follow-up, single-center study showed almost the same effective and safe profile for both Cypher and Taxus stents, except for a better reducing restenosis effect in the Cypher stent. PMID- 20014188 TI - Valvular perforation in left-sided native valve infective endocarditis. AB - BACKGROUND: Left-sided native valve infective endocarditis (LNVIE) can result in mitral (MP) and aortic (AP) valve perforation, the prognostic significance of which remains poorly defined. HYPOTHESIS: Valvular perforation is associated with worse outcomes. METHODS: Retrospective review of patients with LNVIE during 1998 2005 was performed to examine characteristics and outcome predictors of LNVIE complicated by valve perforation. Patients were stratified as: group A: MP or AP detected by transesophageal echocardiography (TEE) or surgery; group B: no TEE evidence of MP or AP. RESULTS: A total of 123 patients were included (group A = 47, group B = 76). In group A, 35 patients (74.5%) had MP alone, 11 (23.4%) had AP alone, and 1 patient had both. Severe valvular insufficiency was encountered more in group A (85.1% versus 59.2%, p = 0.003), so was hemodialysis (40.4% versus 17.1%, p = 0.004) and indications for valvular surgery (93.6% versus 77.6%, p = 0.02). Group A had a higher rate of in-hospital death (31.9% versus 15.8%, p = 0.04). Among patients who had an indication for valvular surgery, the in-hospital mortality rate for those who underwent valvular surgery was 16.7% in group A, and 7.9% in group B (p = 0.4), compared to those who did not undergo surgery (71.4% versus 33.3%, p = 0.04). Amongst survivors, hospital stay was on average 9.2 d longer in group A (38.9 versus 29.7 d, p = 0.05). Univariate analysis revealed association between lower survival and valvular perforation (odds ratio [OR]: 0.4, 95% confidence interval [CI]: 0.17-0.95), that was lost after adjusting for hemodialysis. CONCLUSIONS: Valve perforation complicating LNVIE is associated with hemodialysis, severe valvular insufficiency, and significant morbidity and mortality. Compared to conservative management, early surgical intervention is associated with improved survival. PMID- 20014189 TI - Endocarditis complicating central venous catheter bloodstream infections: a unique form of health care associated endocarditis. AB - BACKGROUND: Endocarditis complicating central venous catheter blood stream infection (CVC-BSI) is a serious complication and is being seen with increasing frequency. METHODS: All patients discharged from our institution with International Classification of Disease (ICD-9) codes of endocarditis and CVC-BSI were identified. The medical records of those meeting our inclusion criteria were reviewed. RESULTS: From October 1, 1998 until December 31, 2006, 24 patients were identified with inpatient mortality of 20.8%. Nine cases were nosocomial and 15 were non-nosocomial. The most common comorbidities were diabetes mellitus (45.8%), chronic kidney disease (58.4%), prior valvular abnormalities (37.5%), and multiple prior hospitalizations (65.2%). There were 13 external lines, 9 tunneled lines, and 2 implantable ports. Responsible microorganisms included Staphylococcus aureus in 54.6%, coagulase-negative staphylococci in 37.5%, Candida species (spp.) in 16.6%, and enterococci in 12.5%. Five cases were polymicrobial. The line tip was within the right atrium (RA) in 37.5%, the superior vena cava (SVC)-RA junction in 20.8%, the SVC in 33.3%, and the pulmonary artery in 4.2% of patients. Sites of endocardial involvement were the aortic valve in 6 patients, mitral valve in 7 patients, tricuspid valve in 6 patients, right atrial wall in 11 patients, and pacemaker wire in 2 patients. Isolated right-sided involvement occurred in 50% of cases, isolated left-sided in 33.4%, and bilateral involvement in 16.6%. Transesophageal echocardiography (TEE) was necessary for diagnosis in 10 cases (41.6%). CONCLUSIONS: Endocarditis complicating CVC-BSI more often involves right-sided structures, with catheter tips in or near the right atrium, frequently requires TEE for diagnosis, and has significant inpatient mortality. PMID- 20014190 TI - Membranous septal aneurysm causing right ventricular outflow tract obstruction. PMID- 20014191 TI - The fourth heart sound-easier to feel than to hear. PMID- 20014192 TI - Giant right coronary aneurysm and syncope. PMID- 20014193 TI - Overexpression of activated nuclear factor-kappa B in aorta of patients with coronary atherosclerosis. AB - BACKGROUND: Inflammation is an established risk factor for atherosclerosis. In an inflammatory state, nuclear factor-kappa B (NF-kappaB) is frequently activated as a key transcription activator for the downstream responses. HYPOTHESIS: The aim of this study was to investigate the changes of NF-kappaB in the aorta of patients with coronary atherosclerosis and its association with atherosclerotic risk factors. METHODS: From 2004 to 2005, we collected a small piece of ascending aorta in the bypass procedure from patients (n = 31) undergoing coronary artery bypass graft (CABG) surgery. The expression of NF-kappaB was determined by immunohistochemistry, and its transcriptional activity was evaluated by electrophoretic mobility shift assay. Celiac aortic tissues from 4 subjects without known atherosclerosis through the kidney donation program were taken as control. RESULTS: NF-kappaB was detectable in aortas from CABG patients with the transcriptional activities significantly increased. The relative level of aortic NF-kappaB expression was elevated in patients who were smokers or with hypertension. Spearman correlation revealed that aortic NF-kappaB expression had significant correlation with coronary severity scores (Gensini score, r = 0.608, P < .05). The NF-kappaB expression was positively correlated with the levels of blood glucose, low-density lipoprotein cholesterol, lipoprotein(a), total cholesterol, and non-high-density lipoprotein cholesterol (P < .05); but negatively correlated with high-density lipoprotein cholesterol (P < .05). CONCLUSIONS: Our study demonstrates a highly activated NF-kappaB in aortas from patients with coronary atherosclerosis, which may reflect overall arterial overinflammatory status. The findings of hyperactive NF-kappaB in aortas may provide a diagnostic parameter for the inflammation that is associated with and may cause atherosclerosis. PMID- 20014194 TI - Letter to the editor. Paclitaxel-Induced ST-segment elevations: A manifestation of allergic acute coronary syndrome (Kounis Syndrome) [corrected]. PMID- 20014195 TI - A rare case of Burkitt's lymphoma presenting as a mediastinal and cardiac mass. PMID- 20014196 TI - Osteoblast adhesion and proliferation on poly(3-octylthiophene) thin films. AB - In this study we assessed the suitability of semiconducting P3OT thin films (30 nm) to sustain attachment, spreading, and proliferation of MC3T3-E1 osteoblasts. Cell area correlated with surface wettability: area was larger on the more hydrophilic surface (TCPS) and lower on the more hydrophobic surface (P3OT). Cells were rounder, characterized by higher circularity values, on TCPS and Si compared to P3OT. P3OT proliferation rate at 24 h fell twofold after 48 h, then recovered at 72 h to a value significantly higher than that on TCPS. Presoaking experiments showed no evidence of cytotoxic effects or leachants from P3OT. Overall, we conclude that P3OT is a viable substrate for osteoblast attachment and proliferation. PMID- 20014197 TI - Light-responsive biomaterials: development and applications. AB - Novel biomaterials are beneficial to the growing fields of drug delivery, cell biology, micro-devices, and tissue engineering. With recent advances in chemistry and materials science, light is becoming an attractive option as a method to control biomaterial behavior and properties. In this Feature Article, we explore some of the early and recent advances in the design of light-responsive biomaterials. Particular attention is paid to macromolecular assemblies for drug delivery, multi-component surface patterning for advanced cell assays, and polymer networks that undergo chemical or shape changes upon light exposure. We conclude with some remarks about future directions of the field. PMID- 20014198 TI - Electrospun fibrous scaffolds with multiscale and photopatterned porosity. AB - The structural and mechanical properties of tissue engineered environments are crucial for successful cellular growth and tissue repair. Electrospinning is gaining wide attention for the fabrication of tissue engineered scaffolds, but the small pore sizes of these scaffolds limit cell infiltration and construct vascularization. To address this problem, we have combined electrospinning with photopatterning to create multiscale porous scaffolds. This process retains the fibrous nature of the scaffolds and permits enhanced cellular infiltration and vascularization when compared to unpatterned scaffolds. This is the first time that photopatterning has been utilized with electrospun scaffolds and is only now possible with the electrospinning of reactive macromers. PMID- 20014199 TI - Exact tests using two correlated binomial variables in contemporary cancer clinical trials. AB - New therapy strategies for the treatment of cancer are rapidly emerging because of recent technology advances in genetics and molecular biology. Although newer targeted therapies can improve survival without measurable changes in tumor size, clinical trial conduct has remained nearly unchanged. When potentially efficacious therapies are tested, current clinical trial design and analysis methods may not be suitable for detecting therapeutic effects. We propose an exact method with respect to testing cytostatic cancer treatment using correlated bivariate binomial random variables to simultaneously assess two primary outcomes. The method is easy to implement. It does not increase the sample size over that of the univariate exact test and in most cases reduces the sample size required. Sample size calculations are provided for selected designs. PMID- 20014200 TI - Percutaneous coronary interventions in nonagenarians: in-hospital mortality and outcome at one year follow-up. AB - BACKGROUND: Limited information is available regarding outcome of very elderly patients referred for percutaneous coronary intervention (PCI). PURPOSE: This study aimed to assess acute and intermediate term clinical outcomes among nonagenarians. METHODS: The study included 32 consecutive nonagenarian patients undergoing PCI between January 2001 to August 2006. There were 6 (19%) patients admitted with acute ST-segment elevation myocardial infarction (STEMI), 10 (31%) patients with non-STEMI, and 16 (50%) patients with unstable angina pectoris. Receiver-operator characteristic curve (ROC) analysis was done to define the relationship between heart rate, blood pressure, left ventricle ejection fraction, serum creatinine level, and mortality. RESULTS: Results: Immediate procedure success was achieved in 28 (88%) patients. Cumulative mortality at hospital discharge was 3(9%), at 6 months it was 6 (19%) and remained 6(19%) at 1 year follow-up. CONCLUSION: Hypotension and low ejection fraction correlated with in-hospital mortality and worst clinical outcome. Procedural success does not appear to decline in nonagenarians. PMID- 20014201 TI - Churg-strauss syndrome presenting with cardiac mass and right ventricular obstruction. AB - A 57-year-old woman presenting with asthma, hypereosinophilia, and generalized unspecific symptoms was diagnosed with Churg-Strauss syndrome. Echocardiography revealed a cardiac mass obliterating the right ventricle and severely impaired left ventricular function, which were caused by endomyocardial fibrosis. Cortisone and cyclophosphamide therapy resulted in amelioration of left ventricular function and significant size reduction of the right ventricular mass. PMID- 20014202 TI - The role of communication and support in return to work following cancer-related absence. AB - OBJECTIVE: Many cancer survivors experience difficulties returning to work. However, there have been relatively few attempts to understand why problems with employer support and work adjustment occur. This paper aims to extend previous work in two ways: first, through exploring the way in which communication and support at work effect cancer survivors on their return to work and during the post-return period; and second, by drawing on a research sample working in the United Kingdom. METHODS: In all, 26 cancer survivors took part in a semi structured telephone interview. Interviews were transcribed and analysed using thematic analysis. RESULTS: The analysis revealed three key findings. First, the central role of communication and support from (and between) occupational health, line managers, and colleagues was highlighted. Second, two discrete processes or periods of return to work were identified: the experience of return to work during the initial period of return and the experiences of post-return to work. Third, during the post-return period, the importance of the delayed impact of cancer on the ability to work, the lack of follow-up and monitoring, and the wear off effect of empathy and support were highlighted as contributing to return-to work difficulties. CONCLUSIONS: This qualitative study highlights the importance of communication within the workplace with regard to the return-to-work process and the need to provide better support and guidance to cancer survivors, line managers and colleagues. Research is required in delineating how employers without occupational health or human resources support manage the return-to-work process. PMID- 20014203 TI - Trimmed weighted Simes' test for two one-sided hypotheses with arbitrarily correlated test statistics. AB - The two-sided Simes test is known to control the type I error rate with bivariate normal test statistics. For one-sided hypotheses, control of the type I error rate requires that the correlation between the bivariate normal test statistics is non-negative. In this article, we introduce a trimmed version of the one-sided weighted Simes test for two hypotheses which rejects if (i) the one-sided weighted Simes test rejects and (ii) both p-values are below one minus the respective weighted Bonferroni adjusted level. We show that the trimmed version controls the type I error rate at nominal significance level alpha if (i) the common distribution of test statistics is point symmetric and (ii) the two-sided weighted Simes test at level 2alpha controls the level. These assumptions apply, for instance, to bivariate normal test statistics with arbitrary correlation. In a simulation study, we compare the power of the trimmed weighted Simes test with the power of the weighted Bonferroni test and the untrimmed weighted Simes test. An additional result of this article ensures type I error rate control of the usual weighted Simes test under a weak version of the positive regression dependence condition for the case of two hypotheses. This condition is shown to apply to the two-sided p-values of one- or two-sample t-tests for bivariate normal endpoints with arbitrary correlation and to the corresponding one-sided p values if the correlation is non-negative. The Simes test for such types of bivariate t-tests has not been considered before. According to our main result, the trimmed version of the weighted Simes test then also applies to the one-sided bivariate t-test with arbitrary correlation. PMID- 20014204 TI - Two brothers with myocardial infarction in the absence of atherosclerotic coronary artery disease: spontaneous coronary thrombosis: case reports of two brothers. AB - Myocardial infarction in the absence of significant atherosclerotic coronary artery disease is not uncommonly encountered in clinical practice. This has been more often seen with the current sensitive biomarker assays for myocardial necrosis. Acute illnesses, spontaneous coronary dissection, sepsis, pulmonary embolism and coagulation disorders are some of the common clinical situation where elevated cardiac markers are noted. We describe two brothers presenting with acute myocardial infarction due to thrombus without any obvious cause. PMID- 20014205 TI - Single-photon emission computed tomography myocardial perfusion imaging in the diagnosis of left main disease. AB - BACKGROUND: Left main disease (LMD), defined as > or = 50% lesion stenosis, occurs in 3% to 5% of patients undergoing catheterization. Limited data on the value of single-photon emission computed tomography (SPECT) imaging for diagnosis of LMD exists. HYPOTHESIS: This study sought to evaluate the diagnostic accuracy of SPECT imaging in the diagnosis of LMD. METHODS: A total of 74 consecutive patients with LMD, identified from our catheterization lab database (January 2003 December 2007) with gated exercise (15 patients) or adenosine (59 patients), thallium 201, or Tc-99m SPECT imaging within 6 months of index angiography were included. Group 1 (Gp 1) included 33 patients with isolated LMD. Group 2 (Gp 2) consisted of 41 patients with LMD and 1-vessel disease (6); LMD and 2-vessel disease (24); and LMD and 3-vessel disease (11). RESULTS: Reversible perfusion defects (PD) were absent in 6 (18%) of Gp 1 and 8 (20%) of Gp 2 patients. Among Gp 1 patients, PD in 1-vessel, 2-vessel, 3-vessel distribution were seen in 20 (61%), 5 (15%), and 2 (6%) patients respectively. In comparison, PD in 1-vessel, 2-vessel, 3-vessel distribution were observed in 20 (49%), 12 (29%), and 1 (2%) of Gp 2 patients respectively. Left main (LM) pattern was noted in 6 patients (Gp 1: 2, Gp 2: 4). Transient ischemic dilatation (TID) was encountered in 34 patients (Gp 1: 17, Gp 2: 17) and in 4 patients with normal perfusion scans. CONCLUSIONS: These data represent the largest analysis of patients with "isolated" LMD. Approximately 19% of patients with LMD have no reversible PD on SPECT. While LM pattern is rare, 1-vessel distribution PD and TID represent the most frequent SPECT abnormalities. PMID- 20014206 TI - Natural history of a giant coronary aneurysm with spontaneous dissection. AB - Primary spontaneous coronary artery dissection may appear in young women during the peripartum period or as a result of atherosclerosis. We present a patient with familial hypercholesterolemia who developed a giant aneurysm of the right coronary artery in the setting of atherosclerotic spontaneous coronary dissection over an 8 year period. This report illustrates the association between spontaneous coronary artery dissection and subsequent coronary aneurysm formation. PMID- 20014207 TI - Neutrophil to lymphocyte ratio as a predictor of long-term mortality in African Americans undergoing percutaneous coronary intervention. AB - BACKGROUND: Neutrophil to lymphocyte ratio (N/L ratio) has been shown to predict long-term mortality in patients undergoing percutaneous coronary intervention (PCI). African Americans have been shown to have lower mean neutrophil counts compared to whites. The usefulness of the N/L ratio in predicting long-term mortality in African Americans undergoing PCI is unknown. METHODS: We evaluated a total of 372 African American patients (327 patients with lower N/L ratio [<3.5] and 45 patients with higher N/L ratio [> or =3.5]) who underwent PCI during January 2003 to August 2005. The primary endpoint was all-cause mortality at a median follow-up to 3.6 years. RESULTS: During the median ( +/- SD) follow-up period of 3.6 +/- 1 years, there were a total of 48 deaths. The mortality rate was 10.4% in the group with a lower N/L ratio and 31.1% in the group with a higher N/L ratio (unadjusted p < 0.001). After adjustment for covariates with significant impact on mortality, N/L ratio was still a strong and independent predictor of long-term mortality with a hazard ratio (HR) of 2.1 (95% confidence interval [CI]: 1.1-4; p = 0.02). N/L ratio was also found to be a strong and independent predictor of long-term mortality even when analyzed as a categorical variable with 3 groups (HR of 0.39 for lower tertile compared to the upper tertile, 95% CI: 0.19-0.81; p = 0.012) and as a continuous variable (p = 0.002). CONCLUSION: N/L ratio is a powerful independent predictor of long-term mortality in African Americans undergoing PCI. PMID- 20014208 TI - Serum N-terminal-pro-brain natriuretic peptide level and its clinical implications in patients with atrial fibrillation. AB - BACKGROUND: Brain natriuretic peptide (BNP) is increasingly being used for screening and monitoring of congestive heart failure. However, the role of BNP in patients with atrial fibrillation (AF) and normal left ventricular function has not been determined. This study investigates serum N-terminal pro-brain natriuretic peptide (NT-proBNP) level and its clinical implications in patients with AF. METHODS: Serum NT-proBNP levels were measured by enzyme-linked immunosorbent assay (ELISA) and transthoracic echocardiography was performed in 136 subjects (90 cases with AF and 46 cases with sinus rhythm [SR]). Subjects were excluded if they had a history of myocardial infarction, cardiomyopathy, rheumatic heart disease, or hyperthyroidism that preceded the onset of AF. Controls (n = 30) were from a healthy outpatient primary care population. Potential determinants of serum NT-proBNP levels were identified by univariate and multivariate analyses. RESULTS: Individuals with AF had higher serum NT proBNP levels (689.56 +/- 251.87 fmol/ml) than those with SR (456.11 +/- 148.14 fmol/ml, P < 0.01) and control subjects (415.83 +/- 62.02 fmol/ml, P < 0.01). Individuals with SR and control subjects did not show significant difference at serum NT-proBNP levels (P > 0.05). The regression model of serum NT-proBNP levels and clinical predictors showed that presence of AF, older age, and larger right atrial diameter were independently predictive of higher serum NT-proBNP values. CONCLUSIONS: Patients with AF were associated with increased serum NT-proBNP levels. Examining the change of serum NT-proBNP levels is helpful to evaluate the cardiac function in patients with AF. PMID- 20014209 TI - Clinical implications of defective B-type natriuretic peptide. AB - Our understanding of the natriuretic peptide system continues to evolve rapidly. B-type natriuretic peptide (BNP), originally thought to be a simple volume regulating hormone that is produced in response to cardiac stretch, has been shown to also play important roles in modulating bronchodilation, endothelial function, and cardiac remodeling. Recent data demonstrate that elevated levels of BNP in patients with heart failure do not represent a simple ratcheting up of normal production in response to increased stimulus. Instead, we now know that chronic stimulation of BNP synthesis induces a reversion to fetal gene expression, resulting in production of high molecular weight forms of BNP that are functionally deficient. Standard point-of-care BNP assays are immunoassays that will detect any molecule containing the target epitopes. Consequently, these assays cannot distinguish between defective, high molecular weight forms of BNP and normal, physiologically active BNP. In 2 separate evaluations, mass spectroscopy detected little, if any, normal BNP in patients with heart failure, despite the appearance of high circulating levels of immunoreactive BNP (iBNP) using commercial assays. Therefore, these commercial assays should be considered to be only an indication of myocardial stress. They do not measure physiologic BNP activity. This accounts for the "BNP paradox," namely, that administration of exogenous recombinant human BNP (rhBNP, nesiritide) has substantial clinical and hemodynamic impact in the presence of high levels of circulating iBNP using commercial assays. In addition to its short-term hemodynamic impact, rhBNP may have other important effects in this setting, and further investigation is warranted. PMID- 20014210 TI - A restrictive inflow pattern does not predict implantable cardioverter defibrillator therapy in primary prevention. AB - BACKGROUND: Current guidelines for the use of implantable cardioverter defibrillators (ICDs) are broad and significantly increase the cost of caring for patients with heart failure. In an effort to identify the specific subset of patients who benefit from this therapy, the predictive value of numerous echocardiographic parameters have been studied. Severe diastolic dysfunction has been shown to predict adverse events in a group of patients who received an ICD for secondary prevention, but has not been tested in those who receive ICDs for primary prevention. HYPOTHESIS: We tested the hypothesis that a restrictive mitral inflow pattern on echocardiography will predict the risk of appropriate therapy in this patient population. METHODS: This retrospective study identified 145 consecutive patients who met primary prevention criteria for ICD implantation and had an echo performed no more than 1 year prior to receiving the ICD. A restrictive pattern was defined as a mitral inflow E/A > 2 or a deceleration time < 150 ms. RESULTS: A restrictive pattern was present in 69 patients (40.7% of the group). Appropriate ICD therapy occurred in 8 (11.5%) subjects with a restrictive pattern and 14 (18.4%) with a nonrestrictive pattern over 680 days of average follow-up (P = not significant). Cox regression analysis showed the presence of a restrictive pattern was not helpful in predicting time to first ICD therapy. CONCLUSIONS: In a population of patients who received ICDs for primary prevention, echocardiographic findings of severe diastolic dysfunction were not helpful in targeting the use of ICDs to those at highest risk. PMID- 20014211 TI - Hemoglobin levels predict exercise performance, ST-segment depression, and outcome in patients referred for routine exercise treadmill testing. AB - BACKGROUND: While the role of hemoglobin in heart failure and renal disease has been investigated, little is known about its effect on clinical exercise test performance and mortality in patients referred for routine exercise treadmill testing (ETT). HYPOTHESIS: Patients with low hemoglobin will have poor exercise capacity and would be at increased risk of mortality and cardiovascular (CV) events. METHODS: Clinical variables, laboratory values, and exercise treadmill data were obtained for 1,799 patients referred for routine ETT from 1997 to 2004. All-cause mortality was obtained from the United States Social Security death index and autopsy reports or clinical notes were used to determine CV events and mortality. P values < 0.05 were considered significant. RESULTS: Our population had a mean age of 58 +/- 12 years, 16% had diabetes, 53% had hypertension, 35% had hypercholesterolemia, and 67% had a history of smoking. During follow-up, 10.3% of patients died, 3.9% of patients died of CV causes, and 11.6% had cardiovascular events. Anemic patients (hemoglobin [Hgb] < 13 g/dL) achieved lower metabolic equivalents (METs) than nonanemic patients and had more ST segment depression (15.5% versus 8.6%, p < 0.004). Proportional hazard analysis demonstrated that hemoglobin was significantly associated with all-cause mortality (p < 0.0007), CV mortality (p < 0.009), and CV events (p < 0.01). Kaplan-Meier survival analysis demonstrated that anemic patients had significantly higher mortality and CV events. CONCLUSION: Hemoglobin is significantly associated with exercise performance, ST-segment depression during ETT, mortality, and cardiovascular events. The incorporation of hemoglobin may add diagnostic and prognostic information to ETT. PMID- 20014212 TI - Patients with metabolic syndrome have prolonged corrected QT interval (QTc). AB - BACKGROUND: Prolongation of corrected QT interval (QTc) increases morbidity and mortality and QTc has been found to be longer in patients with diabetes mellitus than in healthy controls. It is still inconclusive whether the metabolic syndrome results in QTc prolongation. HYPOTHESIS: We hypothesized that metabolic syndrome might contribute to risk of QTc prolongation. The hypothesis was tested in a large population. METHODS: A total of 5,815 individuals (men: 1,950, women: 3,865 aged 20-80 years) were enrolled. Metabolic syndrome was defined according to the revised third National Cholesterol Education Program Adult Treatment Panel (NCEP ATP III). QTc was calculated by using Bazett and Fridericia equations and the corrected JT interval (JTc) was derived by subtracting the QRS duration from the QTcB. All individuals had physical examinations, electrocardiograms, echocardiography, and blood tests. RESULTS: Individuals with metabolic syndrome had longer QTcs and JTc than those without metabolic syndrome (439.84 ms versus 430.90 ms in men, 456.37 ms versus 445.12 ms in women, respectively, p < 0.001 using Bazett formula). The more the number of abnormal metabolic parameters they had, the longer the QTcs and JTc they had. Trend analysis indicated that QTcB, QTcF, and JTc were significantly correlated to the number of abnormal metabolic parameters both in men and in women. After being adjusted for conventional risk factors, QTcB, QTcF, and JTc remained negatively associated with serum potassium concentration and positively associated with interventricular septal thickness. CONCLUSIONS: Metabolic syndrome is a risk factor for prolonged QTc, which may further increase cardiovascular morbidity and mortality in the subjects with metabolic syndrome. PMID- 20014213 TI - Myocardial stunning following combined modality combretastatin-based chemotherapy: two case reports and review of the literature. AB - Myocardial stunning, known as stress cardiomyopathy, broken-heart syndrome, transient left ventricular apical ballooning, and Takotsubo cardiomyopathy, has been reported after many extracardiac stressors, but not following chemotherapy. We report 2 cases with characteristic electrocardiographic and echocardiographic features following combined modality therapy with combretastatin, a vascular disrupting agent being studied for treatment of anaplastic thyroid cancer. In 1 patient, an ECG performed per protocol 18 hours after drug initiation showed deep, symmetric T-wave inversions in limb leads I and aVL and precordial leads V(2) through V(6). Echocardiography showed mildly reduced overall left ventricular systolic function with akinesis of the entire apex. The patient had mild elevations of troponin I. Coronary angiography revealed no epicardial coronary artery disease. The electrocardiographic and echocardiographic abnormalities resolved after several weeks. The patient remains stable from a cardiovascular standpoint and has not had a recurrence during follow-up. An electrocardiogram performed per protocol in a second patient showed deep, symmetric T-wave inversions throughout the precordial leads and a prolonged QT interval. Echocardiography showed mildly reduced left ventricular function with hypokinesis of the apical-septal wall. Acute coronary syndrome was ruled out, and both the electrocardiographic and echocardiographic changes resolved at follow up. Although the patient remained pain-free without recurrence of anginal symptoms during long-term follow-up, the patient developed progressive malignancy and died. PMID- 20014216 TI - Designed DNA crystals: triangles with short sticky ends. PMID- 20014219 TI - Editorial: Biotech in the post genomic era. PMID- 20014218 TI - Nurse staffing and post-surgical complications using the present on admission indicator. AB - We evaluated the relationship between registered nurse (RN) staffing and six post surgical complications: pneumonia, septicemia, urinary tract infections, thrombophlebitis, fluid overload, and decubitus ulcers, in a dataset that contained the present on admission (POA) indicator. We analyzed a longitudinal panel of 283 acute care hospitals in California from 1996 to 2001. Using an adaptation of the Quality Health Outcomes Model, we found no statistically significant relationships between RN staffing and the complications. In addition, the signs of the relationships were opposite to those expected. That is, as staffing increased, so did some of the complications. We discuss potential reasons for these anomalous results, including the possibility that increases in RN staffing may result in earlier detection of complications. Other explanations include issues with risk adjustment, the lack of nurse level variables in the model, and issues with the POA indicator itself. PMID- 20014222 TI - Meeting report: European Congress on Biotechnology 2009. PMID- 20014224 TI - Visual stimulation enhances auditory processing in 3-month-old infants and adults. AB - Behavioral work demonstrates human infants are sensitive to a host of intersensory properties and this sensitivity promotes early learning and memory. However, little is known regarding the neural basis of this ability in infants. Using event-related potentials (ERPs) with infants and adults, we show that during passive viewing auditory evoked brain responses are increased with the presence of simultaneous visual stimulation. Results converge with previous adult neuroimaging studies, single-cell recordings in nonhuman animals, and behavioral studies with human infants to provide evidence for an elevated status of multisensory stimulation in infancy. Furthermore, these results may provide a neural marker of multisensory audio-visual processing in infants that can be used to test developmental theories of how information is integrated across the senses to form a unitary perception of the world. PMID- 20014225 TI - The effect of taste familiarity on intake and taste reactivity in infant rats. AB - With infant rats, unlike with adults, increased intake of a taste after mere exposure to this stimulus is not consistently found; this has sometimes been interpreted as a failure by the immature subject to recognize tastes as familiar. We studied the effect of preexposure to a tastant, measuring taste reactivity and intake in 14-day-old rats. Familiarity increased hedonic response to sucrose, but also increased aversive response to quinine and ethanol. With the sucrose-quinine compound, familiarity increased both the hedonic and the aversive reaction to the stimulus. In no case was a differential reactivity to water observed. Significant increased intake after familiarization was only found with quinine or the sucrose quinine compound. Results indicate that in infant rats, and with the present parameters, taste familiarity enhances responsiveness to these stimuli, an effect not always accompanied by detectable changes in intake. PMID- 20014227 TI - Manufacturing antibodies in the plant cell. AB - Plants have long been considered advantageous platforms for large-scale production of antibodies due to their low cost, scalability, and the low chances of pathogen contamination. Much effort has therefore been devoted to efficiently producing mAbs (from nanobodies to secretory antibodies) in plant cells. Several technical difficulties have been encountered and are being overcome. Improvements in production levels have been achieved by manipulation of gene expression and, more efficiently, of cell targeting and protein folding and assembly. Differences in mAb glycosylation patterns between animal and plant cells are being successfully addressed by the elimination and introduction of the appropriate enzyme activities in plant cells. Another relevant battlefield is the dichotomy between production capacity and speed. Classically, stably transformed plant lines have been proposed for large scale mAb production, whereas the use of transient expression systems has always provided production speed at the cost of scalability. However, recent advances in transient expression techniques have brought impressive yield improvements, turning speed and scalability into highly compatible assets. In the era of personalized medicines, the combination of yield and speed, and the advances in glyco-engineering have made the plant cell a serious contender in the field of recombinant antibody production. PMID- 20014229 TI - Biotech round the world: Iceland. PMID- 20014231 TI - Effect of age-dependent exposure to lead on hepatotoxicity and nephrotoxicity in male rats. AB - Lead is known to induce a broad range of physiological, biochemical, and behavioral dysfunctions in laboratory animals and humans. This includes age specific variations in absorption, retention, and tissue distribution of lead. This study was carried out to investigate the effects of chronic exposure to lead (50 mg/L) on liver and kidneys of two different age groups of male rats treated with lead from delivery until puberty period (40 days) and postpuberty period (65 days). For this purpose, the concentrations of thiobarbituric acid reactive substance (TBARS), total thiol groups (SH), and superoxide dismutase (SOD) activity were measured in the liver and kidney of rats. Renal function was analyzed by determining creatinine, acid uric, and urea. Plasma activities of alanine aminotransferase, aspartate aminotransferase, alkaline phosphatase, and albumin were determined spectrophotometrically to evaluate hepatic function. These markers of damage were determined to assess the level of toxicity in these animals. Our results clearly show that the administration of lead produces oxidative damage in liver and kidney, as strongly suggested by the significant increase in TBARS, decrease in total SH, and the alteration of SOD activity. In young lead-exposed animals, lead-induced perturbations on the synthetic function of the liver and the kidney were more pronounced. However, nephropathy is evident for adult lead-exposed animals. It is concluded that lead induces severe hepatic and renal toxicity, which depends on the age of the animals and the target organ. PMID- 20014232 TI - International amphibian micronucleus standardized procedure (ISO 21427-1) for in vivo evaluation of double-walled carbon nanotubes toxicity and genotoxicity in water. AB - Considering the important production of carbon nanotubes (CNTs), it is likely that some of them will contaminate the environment during each step of their life cycle. Nevertheless, there is little known about their potential ecotoxicity. Consequently, the impact of CNTs on the environment must be taken into consideration. This work evaluates the potential impact of well characterized double-walled carbon nanotubes (DWNTs) in the amphibian larvae Xenopus laevis under normalized laboratory conditions according to the International Standard micronucleus assay ISO 21427-1:2006 for 12 days of half-static exposure to 0.1-1 10 and 50 mg L(-1) of DWNTs in water. Two different endpoints were carried out: (i) toxicity (mortality and growth of larvae) and (ii) genotoxicity (induction of micronucleated erythrocytes). Moreover, intestine of larvae were analyzed using Raman spectroscopy. The DWNTs synthetized by catalytic chemical vapor deposition (CCVD) were used as produce (experiment I) and the addition of Gum Arabic (GA) was investigated to improve the stability of the aqueous suspensions (experiment II). The results show growth inhibition in larvae exposed to 10 and 50 mg L(-1) of DWNTs with or without GA. No genotoxicity was evidenced in erythrocytes of larvae exposed to DWNTs, except to 1 mg L(-1) of DWNTs with GA suggesting its potential effect in association with DWNTs at the first nonacutely toxic concentration. The Raman analysis confirmed the presence of DWNTs into the lumen of intestine but not in intestinal tissues and cells, nor in the circulating blood of exposed larvae. PMID- 20014233 TI - Therapy for lysosomal storage disorders. AB - In the last years, much progress has been achieved in the field of lysosomal storage disorders. In the past, no specific treatment was available for the affected patients; management mainly consisted of supportive care and treatment of complications. As orphan drug regulations, however, encouraged development of drugs for these disorders by granting marketing exclusivity for 10 years and other commercial benefits, enzyme replacement therapy became available for lysosomal storage disorders, such as Gaucher disease, Fabry disease, mucopolysaccharidoses type I, II, and VI, and Pompe disease. This review will summarize the efficacy and clinical status of hematopoietic stem cell transplantation, enzyme replacement, and substrate deprivation therapy, and describe new therapeutic perspectives currently under preclinical investigations such as chaperone-mediated therapy, stop-codon read-through therapy, and gene therapy. PMID- 20014234 TI - Mycobacterial stress regulation: The Dps "twin sister" defense mechanism and structure-function relationship. AB - In this work, we have tried to emphasize the connection between mycobacterial growth and regulation of gene expression. Utilization of multiple carbon sources and diauxic growth helps bacteria to regulate gene expression at an optimum level so that the inhospitable conditions encountered during nutrient depletion can be circumvented. These aspects will be discussed with respect to mycobacterial growth in subsequent sections. Identification and characterization of genes induced under such conditions is helpful to understand the physiology of the bacterium. Although it is necessary to compare the total expression profile of proteins as they transit from vegetative growth to stationary phase, at times a lot of insights can be deciphered from the expression pattern of one or two proteins. We have compared the protein expression and sigma factor selectivity of two such proteins in M. smegmatis to understand the differential regulation of genes playing diverse function in the same species. Some newer insights on the structure and function of one of the Dps proteins are also explained. PMID- 20014235 TI - Micro-RNA-375 inhibits lung surfactant secretion by altering cytoskeleton reorganization. AB - Lung surfactant is secreted via exocytosis of lamellar bodies from alveolar epithelial type II cells. Whether micro-RNAs regulate lung surfactant secretion is unknown. Micro-RNA-375 (miR-375) has been shown to be involved in insulin secretion. In this article, we report that the overexpression of miR-375 inhibited lung surfactant secretion. However, miR-125a, miR-30a, miR-1, miR-382 and miR-101 did not influence lung surfactant secretion. miR-375 had no effects on surfactant synthesis or the formation of lamellar bodies. However, miR-375 did abolish the lung surfactant secretagogue-induced disassembly and reassembly of cytoskeleton. Our results suggest that miR-375 regulates surfactant secretion via the reorganization of cytoskeleton. PMID- 20014236 TI - Dissecting the signal transduction pathways triggered by galectin-glycan interactions in physiological and pathological settings. AB - Galectins are a family of evolutionarily conserved animal lectins with pleiotropic functions and widespread distribution. Fifteen members have been identified in a wide variety of cells and tissues. Through recognition of cell surface glycoproteins and glycolipids, these endogenous lectins can trigger a cascade of intracellular signaling pathways capable of modulating cell differentiation, proliferation, survival, and migration. These cellular events are critical in a variety of biological processes including embryogenesis, angiogenesis, neurogenesis, and immunity and are substantially altered during tumorigenesis, neurodegeneration, and inflammation. In addition, galectins can modulate intracellular functions and this effect involves direct interactions with distinct signaling pathways. In this review, we discuss current knowledge on the intracellular signaling pathways triggered by this multifunctional family of beta-galactoside-binding proteins in selected physiological and pathological settings. Understanding the "galectin signalosome" will be essential to delineate rational therapeutic strategies based on the specific control of galectin expression and function. PMID- 20014237 TI - Various aspects of diagnostic quantification. PMID- 20014239 TI - Changes will not be made just for the sake of changing. PMID- 20014238 TI - Exercise echocardiography in asymptomatic survivors of childhood cancer treated with anthracyclines: a prospective follow-up study. AB - BACKGROUND: Exercise echocardiography reveals abnormalities in asymptomatic childhood cancer survivors who previously have been treated with anthracyclines. We determined the added value of monitoring childhood cancer survivors with exercise echocardiography compared to monitoring with resting echocardiography alone to predict anthracycline-induced cardiotoxicity. Secondary aims were to evaluate change in resting cardiac function over 10 years and to determine risk factors for late cardiotoxicity. PROCEDURE: We invited a cohort of 110 originally asymptomatic anthracycline-treated childhood cancer survivors, who had undergone cardiac tests including exercise echocardiography 10.5 years earlier, for new cardiac evaluation. Each subject underwent a resting echocardiogram at both evaluations. At first evaluation a repeat echocardiogram was performed following peak exercise. Resting echocardiographic parameters were converted to z character. RESULTS: Ninety-two of 110 survivors (mean anthracycline dose 307 mg/m(2), mean follow-up time from start of treatment 8.2 years at first and 18.8 years at second evaluation) were evaluated prospectively. Mean resting fractional shortening z-character (RFSz) decreased from -0.18 to -0.93. Higher cumulative anthracycline dose was a risk factor for a lower RFSz at late follow-up (P = 0.0002). Adding exercise fractional shortening (XFS) to a model containing RFSz did not improve prediction of abnormal RFSz at late follow-up. CONCLUSIONS: Monitoring with exercise echocardiography has no added value to monitoring with resting echocardiography alone in predicting late anthracycline-induced cardiotoxicity in childhood cancer survivors. RFSz deteriorates over time, even in originally asymptomatic patients. Previous treatment with higher cumulative anthracycline dose is the main risk factor for a lower RFSz at late follow-up. PMID- 20014241 TI - Crystallographic and mass spectrometric analyses of a tandem GNAT protein from the clavulanic acid biosynthesis pathway. AB - (3R,5R)-Clavulanic acid (CA) is a clinically important inhibitor of Class A beta lactamases. Sequence comparisons suggest that orf14 of the clavulanic acid biosynthesis gene cluster encodes for an acetyl transferase (CBG). Crystallographic studies reveal CBG to be a member of the emerging structural subfamily of tandem Gcn5-related acetyl transferase (GNAT) proteins. Two crystal forms (C2 and P2(1) space groups) of CBG were obtained; in both forms one molecule of acetyl-CoA (AcCoA) was bound to the N-terminal GNAT domain, with the C-terminal domain being unoccupied by a ligand. Mass spectrometric analyzes on CBG demonstrate that, in addition to one strongly bound AcCoA molecule, a second acyl-CoA molecule can bind to CBG. Succinyl-CoA and myristoyl-CoA displayed the strongest binding to the "second" CoA binding site, which is likely in the C terminal GNAT domain. Analysis of the CBG structures, together with those of other tandem GNAT proteins, suggest that the AcCoA in the N-terminal GNAT domain plays a structural role whereas the C-terminal domain is more likely to be directly involved in acetyl transfer. The available crystallographic and mass spectrometric evidence suggests that binding of the second acyl-CoA occurs preferentially to monomeric rather than dimeric CBG. The N-terminal AcCoA binding site and the proposed C-terminal acyl-CoA binding site of CBG are compared with acyl-CoA binding sites of other tandem and single domain GNAT proteins. PMID- 20014242 TI - Stereoselective and multiple carrier-mediated transport of cetirizine across Caco 2 cell monolayers with potential drug interaction. AB - The aim of this study was to explore potential transport mechanisms of cetirizine enantiomers across Caco-2 cells. Cetirizine displayed polarized transport at concentrations ranging from 4.0 to 80.0 microM, with the permeability in the secretory direction being 1.4- to 4.0-fold higher than that in the absorptive direction. Cetirizine enantiomers were transported distinctively different from each other. In the presence of inhibitors of P-glycoprotein (P-gp) and multidrug resistance-associated protein (MRP), the absorptive transport was enhanced and secretory efflux was diminished. When verapamil, indomethacin, or probenecid were present, the difference in the absorptive permeability of R-cetirizine and S cetirizine substantially intensified, whereas quinidine could eliminate. R cetirizine significantly increased the efflux ratio of rhodamine-123 and doxorubicin in a fashion indicative of the upregulation of P-gp and MRP activities. However, S-cetirizine played a role of an inhibitor for P-gp and MRP. Ranitidine modified the absorption of cetirizine enantiomers, suggesting that the potential drug-drug interaction would significantly change the cetirizine pharmacokinetics. In conclusion, the results indicated that there are several efflux transporters including P-gp and MRP participating the absorption and efflux of cetirizine, which showed enantioselectivity in the transmembrane process. In addition, both P-gp and MRP functions could be modulated by cetirizine in chiral discriminative ways. PMID- 20014243 TI - Liquid chromatographic direct resolution of flecainide and its analogs on a chiral stationary phase based on (+)-(18-crown-6)-2,3,11,12-tetracarboxylic acid. AB - Flecainide, an antiarrythmic agent, and its analogs were resolved on a high performance liquid chromatographic chiral stationary phase (CSP) based on (+)-(18 crown-6)-2,3,11,12-tetracarboxylic acid with the use of a mobile phase consisting of methanol-acetonitrile-trifluoroacetic acid-triethylamine (80/20/0.1/0.3, v/v/v/v). The chiral resolution was quite successful, the separation factors (alpha) and the resolutions (R(S)) for 20 analytes including flecainide being in the range of 1.19-1.82 and 1.73-6.80, respectively. The ortho-substituent of the benzoyl group of analytes was found to cause decrease in the retention times of analytes probably because of the conformational deformation of analytes originated from the steric hindrance exerted by the ortho-substituent. PMID- 20014244 TI - Electrostatic chiral distinction: tetrahedral model dimers. AB - Although chiral distinction plays a pervasive role in chemistry, a complete understanding of how this takes place is still lacking. In this work, we expand the earlier described minimal requirement of so called four-point interactions (vide infra). We focus on chiral point charge model systems as a means to aid in the dissection of the underlying, operative principles. We also construct models with defined symmetry characteristics. By considering extensive constellations of diastereomeric complexes, we are able to identify emerging principles for chiral distinction. As previously postulated, all the diastereomeric complexes, regardless of their nominal contact-points, possess a chiral distinction energy. In the comparison of complexes, we find that, contrary to chemical intuition, the magnitude of chiral distinction does not correlate with the stability of the complexes, i.e., consideration of low energy complexes may not be an effective way to evaluate chiral distinction. Similarly, we do not find a correlation between the number of contact-points and chiral distinction. Moreover, favorable interactions and facile chiral distinction appear to be unrelated. We also see some tendency for greater chiral distinction in less symmetric systems, although this may not be general. These studies can now form the basis to fold in higher levels of complexity into the models so as to gain further insights into the nature of chiral distinction. PMID- 20014245 TI - Polarization control of ultrashort mid-IR laser pulses for transient vibrational circular dichroism measurements. AB - Linear dichroism and birefringence artifacts are a major source of concern in transient circular dichroism measurements. They mainly arise from interaction of an imperfectly circular polarized probe beam with a nonisotropic sample. We present in this article a procedure to generate mid-IR pulses of highly symmetric left and right handed circular or elliptical polarization states for transient VCD measurements. An infrared femtosecond laser source is synchronized to the natural frequency of a photo elastic modulator. Residual static birefringence of the modulator and the sample cell can be largely compensated by carefully controlling the arrival time of the ultrashort probe pulses at the modulator. PMID- 20014246 TI - Myokymic discharges in amyotrophic lateral sclerosis (ALS): a rare electrophysiologic finding? AB - Myokymic discharges (MDs) are uncommonly recognized in amyotrophic lateral sclerosis (ALS). The electrophysiologic findings in 96 ALS patients were retrospectively reviewed. MDs were found in 5.2% of patients, in 0.81% of total muscles examined (8.2% cranial muscles vs. 0.15% limb muscles). The higher frequency of MDs in cranial muscles suggests a difference in the metabolic environment or other mechanism of instability of the anterior horn cells in the brainstem compared to the spinal cord. PMID- 20014247 TI - Laser-assisted liposuction using a novel blend of lipid- and water-selective wavelengths. AB - BACKGROUND AND OBJECTIVES: Potential benefits of laser-assisted liposuction include enhanced body shaping and skin tightening with reduced patient downtime. In this clinical study, we investigate whether additional safety and efficacy are achieved with a diode laser system that includes a novel 924 nm lipid-selective wavelength. STUDY DESIGN/MATERIALS AND METHODS: Clinical subjects were treated with a continuous-wave (CW) power 924/975 nm diode laser system in the submental area, abdomen, thighs and flanks. Side effects were documented. Efficacy evaluation included investigator-assessed improvements and subject self assessments. RESULTS: Study investigators observed good to excellent improvement across all subjects (n = 19) by 3 months post-treatment in all four categories evaluated. Facility of tip advancement was rated as "extremely easy" or "easy" in 83% of rated treatments. "Good" to "excellent" skin textural improvement was observed in 83% of rated subjects. Additional benefits included reduced operator fatigue and uniformity of treatment. Patients tolerated the procedure well with minimal and transient side effects. Immediate side effects of erythema, bruising and edema were graded as mild or trace. No long-term side effects were observed. Seventy-eight percent of all subjects were extremely satisfied with the procedure and 89% would do the procedure again. Eighty-nine percent of subjects felt their appearance of unwanted fat was significantly improved with smoother and tighter skin (72%), and 94% would recommend the treatment to family and friends. CONCLUSION: These clinical findings demonstrate that the 924/975 nm diode laser system provides safe and efficacious body shaping, fat reduction and enhanced skin tightening. Ease of movement through the tissue of both the laser system treatment tip and the aspiration cannula resulted in reduced fatigue for the treating physician. Patient benefits included high patient satisfaction, significant reductions in fat, and smoother and tighter skin with minimal downtime. PMID- 20014250 TI - Superior lipolytic effect of the 1,444 nm Nd:YAG laser: comparison with the 1,064 nm Nd:YAG laser. AB - BACKGROUND AND OBJECTIVES: Recently developed laser lipolysis systems have been disappointing because they require more time to remove the same amount of fat than other liposuction methods. A new Nd:YAG laser has been introduced that uses the 1,444 nm wavelength, better absorbed by fat. STUDY DESIGN/MATERIALS AND METHODS: This study consisted of two protocols. The first protocol was an in vivo minipig model. Four 10x10 cm(2) areas were treated on the back of the first minipig. Using the same total energy and power settings (5,000 J, 8 W), both the 1,064 nm and 1,444 nm lasers were used to irradiate the two cephalic areas. The two caudal areas were irradiated with both lasers, using the maximum power settings (12 W with the 1,064 nm laser, 8 W with the 1,444 nm laser). Another minipig was administered a preoperative injection of tumescent solution and treated with the same condition. Measurements of fat volume with computed tomography and histologic exams were conducted. The second experiment involved in vitro human fat. Equal amounts of human fat, harvested by liposuction, were put into test tubes and irradiated with 1,064 nm and 1,444 nm lasers. Oil production was measured from each test tube. RESULTS: A marked reduction in fat volume and more oil vacuoles and giant cells in histology were identified with the 1,444 nm wavelength compared to the 1,064 nm wavelength. Human fat in the in vitro experiments also revealed more oil production following the use of the 1,444 nm laser. CONCLUSION: The 1,444 nm Nd:YAG laser showed a greater lipolytic effect compared to the 1,064 nm Nd:YAG laser in in vivo minipig and in vitro human fat experiments. To achieve a full understanding of the effects of 1,444 nm Nd:YAG laser lipolysis on the human body, in vivo experimentation will be necessary. PMID- 20014251 TI - Discussion of laser-assisted liposuction. AB - There has been a significant advancement in the use of lasers for body contouring over the past several years. This article will review the recent past and discuss the future of this trend. INTRODUCTION: There is much confusion over the definition of procedures using lasers for lipolysis and liposuction. This review article discusses the definitions of suction-assisted liposuction, laser lipolysis, and laser-assisted liposuction in detail. The development of these procedures is then discussed and pertinent articles are reviewed. MATERIALS AND METHODS: Pubmed and FDA.gov web sites were searched for published articles and FDA approved devices employing lasers for targeting fat for body contouring. This information along with the authors' personal experience was used to review and discuss this topic. DISCUSSION: There is a specific difference between suction assisted liposuction, laser-assisted liposuction, and laser lipolysis. This review article clarifies these definitions and highlights the recent articles employing lasers for minimally invasive fat reduction. PMID- 20014252 TI - Non-invasive cryolipolysis for subcutaneous fat reduction does not affect serum lipid levels or liver function tests. AB - BACKGROUND AND OBJECTIVE: Cryolipolysis provides a method of non-invasive fat reduction that significantly reduces subcutaneous fat without injury to adjacent tissues. Preliminary animal and human data have suggested that cryolipolysis has no effect on serum lipid profiles or liver tests. This study was intended to more fully document any effect of this procedure on lipid and liver-related blood tests. STUDY DESIGN/MATERIALS AND METHODS: Forty subjects with fat bulges on their flanks ("love handles") were treated bilaterally with a non-invasive device (Zeltiq Aesthetics, Pleasanton, CA) that precisely cools tissue to achieve a reduction in the fat layer. Serum lipid levels and liver tests were measured prior to treatment, and at 1 day and 1, 4, 8, and 12 weeks post-treatment. RESULTS: No meaningful changes in mean values were observed for any blood lipid level or liver test at any point over the 12-week follow-up period. CONCLUSION: Cryolipolysis, when used for reduction of subcutaneous flank fat, is not associated with changes in serum lipids or liver test results. PMID- 20014253 TI - Low-level laser therapy as a non-invasive approach for body contouring: a randomized, controlled study. AB - BACKGROUND AND OBJECTIVE: Transmission electron microscopic images have demonstrated the formation of transitory pores in adipocyte cell membranes followed by the collapse of adipose cells subsequent to laser irradiation of 635 nm. The objective is to evaluate the application of a 635 nm and 17.5 mW exit power per multiple diode laser for the application of non-invasive body contouring of the waist, hips, and thighs. STUDY DESIGN/PATIENTS AND METHODS: Double-blind, randomized, placebo-controlled trial of a 2-week non-invasive laser treatment conducted from May 2007 to June 2008 across multiple-private practice sites in the United States of America. Sixty-seven volunteers between the ages of 18-65 with a body mass index (BMI) between 25 and 30 kg/m(2) and who satisfied the set inclusion criteria participated. Eight of the 67 subjects did not have circumference measurements recorded at the 2-week post-procedure measurement point. Participants were randomly assigned to receive low-level laser treatments or a matching sham treatment three times per week for 2 weeks. Reduction in the total combined inches of circumference measurements of the waist, hip and bilateral thighs from baseline to the completion of the 2-week procedure administration phase was assessed. RESULTS: Participants in the treatment group demonstrated an overall reduction in total circumference across all three sites of -3.51 in. (P < 0.001) compared with control subjects who revealed a -0.684 reduction (P < 0.071745). Test group participants demonstrated a reduction of 0.98 in. (P < 0.0001) across the waist, -1.05 in. (P < 0.01) across the hip, and 0.85 in. (P < 0.01) and -0.65 in. (P < 0.01) across the right and left thighs from baseline to 2 weeks (end of treatment). At 2 weeks post-procedure, test group subjects demonstrated a gain of 0.31 total inches collectively across all three sites. CONCLUSION: These data suggest that low-level laser therapy can reduce overall circumference measurements of specifically treated regions. PMID- 20014254 TI - A pilot study of the efficacy of a 1,064 and 1,320 nm sequentially firing Nd:YAG laser device for lipolysis and skin tightening. AB - BACKGROUND: Laser-assisted lipolysis with a medium pulsed 1,064 nm Neodymium:Yttrium-Aluminum-Garnet (Nd:YAG) system has been used since FDA approval in October 2006 [1]. Since then, this technology has been advanced to include an additional wavelength (1,320 nm) and an accelerometer designed to improve efficacy and safety. OBJECTIVE: (1) Evaluate the efficacy and safety of a sequentially firing 1,064 and 1,320 nm Nd:YAG laser device for lipolysis. (2) Evaluate the skin tightening effect by photographic documentation and skin measurements. (3) Assess new collagen formation by histologic and scanning electron microscopic studies. METHODS: Twenty subjects with unwanted local adiposities and skin laxity were enrolled. An Nd:YAG laser with sequentially firing wavelengths of 1,064/1,320 nm was used to treat localized areas of body adiposities. Digital photographs were taken before and after treatment, blinded independent observers graded improvement utilizing a percentile evaluation scale and subjects performed self-assessments. Five of the 20 subjects had the following tests performed: (1) Placement of 4 cmx4 cm square India Ink tattoos for measurement of skin tightening, (2) histology and electron microscopy, (3) biopsies prior to the procedure, 3 days and 1 month after the procedure to determine the presence of new collagen markers. RESULTS: Results showed reduction in localized adiposities with no adverse events from use of this device. Independent observers found 76-100% improvement in adiposities in 85% of subjects and 51-75% improvement in 15% of subjects. Of the subset of five patients, India Ink tattoo maps demonstrated an 18% decrease in surface area indicating a significant skin tightening effect. Histology by H&E, Methylene blue stains, and electron microscopy indicated new collagen formation compared to baseline. CONCLUSION: The 1,064 nm Nd:YAG and 1,320 nm Nd:YAG sequentially firing device with an accelerometer appears to be an effective and safe treatment for localized adiposities with the additional benefit of skin tightening. PMID- 20014255 TI - Three-dimensional surface imaging for clinical trials: improved precision and reproducibility in circumference measurements of thighs and abdomens. AB - BACKGROUND AND OBJECTIVES: Numerous body contouring and laser-assisted liposuction devices have recently obtained or are seeking FDA clearance, and assessment of efficacy of these devices is largely based upon qualitative comparisons of before and after photographs and measurable changes in patient circumference. The current standard for measuring body circumference in clinical trials involves the use of a standard measuring tape. These manual measurements introduce human error and may incorrectly support or refute a device or procedure's efficacy. A promising alternative to manual measurements is three dimensional (3D) photography. This technology allows circumference measurements to be performed on 3D digital models. Our objective is to compare the precision and reproducibility of manual versus 3D photographic measurement of body circumference. STUDY DESIGN/MATERIALS AND METHODS: Thirty subjects completed this IRB approved study. Each subject's thighs and abdomen were measured twice by each of two blinded investigators and twice by the 3D system. RESULTS: For right and left thigh circumference, the variance of the replication errors [measurement 1 measurement 2] for the human investigators was 20.5% larger than the variance for the 3D photography system. For abdominal circumference, the variance for the human investigators was 231.3% larger than the variance for the 3D system. CONCLUSIONS: 3D photography is a valuable tool that enables investigators to reliably detect minute changes in body shape; consequently, 3D photography reduces the number of subjects needed to sufficiently power a clinical study. For studies involving abdominal circumference, utilizing 3D photography reduces the number of subjects needed by 1/2 to 2/3. PMID- 20014256 TI - The role of laser tunnels in laser-assisted lipolysis. AB - BACKGROUND AND OBJECTIVES: Laser-assisted lipolysis (LAL) devices are used as adjuncts to liposuction that create laser tunnels to heat the adipose and connective tissue. Available systems vary significantly across choice of wavelengths, power delivery, and tip design. Rationale are developed for optimum laser parameters evaluated with physical principles and in controlled ex vivo tests. STUDY DESIGN/MATERIALS AND METHODS: A computer model for radiation propagation, thermal conduction and coagulation was developed to study laser tunnels formed in human adipose tissue. An ex vivo study with porcine tissue compared laser tunnels created by a device that operates in short-pulse mode with a 0.6 mm diameter fiber emitting lipid non-selective laser wavelengths to a device that operates in continuous-wave (CW) mode with a 1.5 mm diameter fiber emitting lipid- and water-selective laser wavelengths. RESULTS: Photothermolytic heating is the optimum mechanism to control delivery of heat to the tissue. Fiber tip surface power density can be optimized for ease of penetration and good volumetric heating while avoiding extremely high peak temperatures. CW rather than pulsed laser emission also minimizes peak temperature rise that can interfere with tunnel formation. Lipid- or water-selective laser wavelengths with low absorption yield lower peak temperatures and more uniform volume heating, while lipid-selective wavelengths offer greater safety near the dermis. Ex vivo histology demonstrated greater volumetric heating with the CW, lipid-selective device at similar power settings. CONCLUSION: Wavelength, power delivery, and tip design are based on physical principles and together with treatment technique laser tunnel dimensions can be optimized as confirmed in ex vivo histology. The resulting thermal zones provide ease of penetration through adipose tissue and enable treatment uniformity. Based upon principles of fractional skin treatment the thermal zones induce healing responses in adipose tissue with potential to enhance clinical efficacy. PMID- 20014257 TI - Injectable treatments for adipose tissue: terminology, mechanism, and tissue interaction. AB - BACKGROUND: Just as injectable fillers have addressed the need for non-surgical methods to restore desired volume, a number of injectable therapies purpor to play a comparable role to reduce undesired volume. OBJECTIVE: To review published literature on the history, mechanism of action, and tissue interaction of injectable methods that aim to reduce localized collections of fat. RESULTS: Mesotherapy is an injection technique that has medical and cosmetic applications and is often confused with injectable fat loss therapies; injection lipolysis describes non-ablative fat reduction with agents (such as beta-agonists) that activate adipocyte lipolytic pathways; and adipolytic therapy using biologic detergents (such as deoxycholate) leads to permanent adipocyte ablation. None of these therapies have been cleared for use in fat reduction by any regulatory authority worldwide. CONCLUSIONS: The mechanism of action and tissue effects of injectable fat reducing compounds are diverse but are becoming increasingly understood. PMID- 20014258 TI - A comparison of three separate clinical studies evaluating the safety and efficacy of laser-assisted lipolysis using 1,064, 1,320 nm, and a combined 1,064/1,320 nm multiplex device. AB - BACKGROUND: Laser-assisted lipolysis has been suggested to augment traditional liposculpture by improving skin laxity and providing hemostasis. Previous studies have reported improved hemostasis and smoother post-operative appearance with the 1,064 Nd:YAG laser-assisted lipolysis system. METHODS: Three separate pilot studies were performed. In the first study, both arms were treated with tumescent liposculpture. One arm was randomized to treatment with a subcutaneous 1,064 nm Nd:YAG laser. The second study treated multiple sites with half of the area randomized to receive the 1,064 nm versus the 1,320 nm system followed by aspiration at equal power. The third study treated patients using a combined 1,064/1,320 nm multiplex laser system at multiple sites. The endpoint of laser treatment was determined by an external skin surface temperature of 40 degrees C. In all three studies, photographs were compared at 1 week, 1 month, and 3 months post-operatively. RESULTS: In the first study, no significant improvement over tumescent liposculpture alone was noted using the 10 W, 1,064 nm laser. The second study showed no difference using the 10 W, 1,320 nm versus the 10 W, 1,064 nm laser-assisted lipolysis system. Finally, the multiplex 1,064/1,320 nm system appeared to show improvement in skin laxity and fat reduction. Complications included intra-operative thermal burns in 2 of 20 patients using the multiplex system. No complications were noted using the 1,064 or 1,320 nm 10 W systems. CONCLUSIONS: Laser-assisted lipolysis provides an innovative way to address the problem of skin laxity and fat reduction. Clinical results increased dramatically with the combined 1,064/1,320 nm multiplex system. However, caution should be used when exceeding external skin temperatures of 40 degrees C to avoid unwanted thermal burns. Future studies comparing the end temperature and wavelengths independently may help to conclusively delineate the optimal system. PMID- 20014259 TI - Improvement in arm and post-partum abdominal and flank subcutaneous fat deposits and skin laxity using a bipolar radiofrequency, infrared, vacuum and mechanical massage device. AB - BACKGROUND AND OBJECTIVES: Skin laxity of the body is a growing cosmetic concern. Laxity can result from chronological or photoaging and changes in body dimensions during pregnancy or weight loss. The end result is loose, sagging skin, and localized fat deposits. Liposuction and abdominoplasty or brachioplasty are established approaches to these issues. Patient desire for alternatives to surgical correction has spawned the development of non-invasive body contouring devices. The combination of infrared light (IR), bipolar radiofrequency (RF), vacuum and mechanical massage (Velashape, Syneron Medical Ltd, Israel) has demonstrated efficacy in improving skin appearance and circumference of the thighs [Goldberg et al., Derm Surg 2008; 34:204-209; Fisher et al., Derm Surg 2005; 31:1237-1241; Arnoczky and Aksan, J Am Acad Orthop Surg 2000; 8:305-313; Alster and Tanzi, J Cosmetic Laser Therapy 2005; 7:81-85; Wanitphakdeedecha and Manuskiatti, J Cosmet Dermatol 2006; 5:284-288; Nootheti et al., Lasers Surg Med 2006; 38: 908-912], but only anecdotal evidence has supported its use on other anatomic locations. This study was designed to evaluate the efficacy and safety of Velashape on additional body sites and more rigorously examine the technology's impact on upper arm as well as abdominal and flank circumference. STUDY DESIGN AND METHODS: Subjects were 28-70 years old, skin types I-V. Nineteen subjects underwent 5 weekly treatments of the upper arms, and 10 subjects underwent 4 weekly treatments of the abdomen and flanks. Treatments were performed using Velashape. Circumference measurements, photographs, and subject weights were performed prior to treatment and at 1- and 3-month follow-ups. Subjects were asked to record their treatment satisfaction level. RESULTS: Change in arm circumference, at the 5th treatment was statistically significant with a mean loss of 0.625 cm. At 1- and 3-month follow-ups, mean loss was 0.71 and 0.597 cm respectively. Reduction of abdominal circumference at 3rd treatment was statistically significant with a 1.25 cm mean loss. At 1- and 3-month follow-ups, average loss was 1.43 and 1.82 cm respectively. CONCLUSIONS: This study demonstrates with statistical significance, sustainable reduction in circumference and improvement in appearance of arms and abdomen following treatment with Velashape. PMID- 20014260 TI - A high-throughput model for fat graft assessment. AB - BACKGROUND AND OBJECTIVES: Current fat-graft animal models require weeks, to months, for results. The purpose of this study was to develop a model for the rapid identification of adipocyte protectants, using apoptosis-specific fluorescence. The goal of our model was to predict long-term fat graft survival within a 10-day period. STUDY DESIGN/MATERIALS AND METHODS: Human fat was obtained from liposuction aspirates, washed with saline, and centrifuged at 200g. The fat was then treated with one of four agents: P188, polyethylene glycol (PEG 8000), lipoic acid (LA), or saline control. Fat lobules were explanted over a 10 day period, and then at six weeks. The explanted fat was measured for apoptosis. Samples were weighed, sent for histology, measured for DNA content, and examined using confocal microscopy. RESULTS: Fat-grafts demonstrated variable apoptosis over the 10-day period. P188 and LA treated samples demonstrated 11-28% less apoptosis during early engraftment than saline treated controls. This early reduction in apoptosis correlated to a approximately 20% reduction in reabsorption by weight six weeks later. P188 and LA samples demonstrated three times higher DNA content by PICO green analysis when compared to saline controls. PEG 8000 treated samples demonstrated 11% more apoptosis than saline. PEG 8000 treated samples demonstrated an approximately 10% higher level of reabsorption by weight, and two-times higher levels of DNA. Histology of treated samples at six weeks showed architecturally normal fat in P188 and LA treated fat; whereas PEG 8000 had high levels of inflammatory infiltrates, and saline had large amounts of fibrosis. CONCLUSIONS: This model of fat-grafting and early apoptosis can be used to screen agents and grafting methods and predict long-term graft survival. We show that levels of apoptosis within ten days correlate with weight, DNA, and histology, at six weeks. Using this model, long-term adipocyte survival and graft take can be predicted during the first 10 days post-implantation. PMID- 20014261 TI - The use of focused ultrasound for non-invasive body contouring in Asians. AB - INTRODUCTION: Previous studies demonstrated that multiple treatments using focused ultrasound can be effective as an non-invasive method for reducing unwanted localized fat deposits. The objective of the study is to investigate the safety and efficacy of this focused ultrasound device in body contouring in Asians. METHOD: Fifty-three (51 females and 2 males) patients were enrolled into the study. Subjects had up to three treatment sessions with approximately 1-month interval in between treatment. Efficacy was assessed by changes in abdominal circumference, ultrasound fat thickness, and caliper fat thickness. Weight change was monitored to distinguish weight loss induced changes in these measurements. Patient questionnaire was completed after each treatment. The level of pain or discomfort, improvement in body contour and overall satisfaction were graded with a score of 1-5 (1 being the least). Any adverse effects such as erythema, pain during treatment or blistering were recorded. RESULT: The overall satisfaction amongst subjects was poor. Objective measurements by ultrasound, abdominal circumference, and caliper did not show significant difference after treatment. There is a negative correlation between the abdominal fat thickness and number of shots per treatment session. CONCLUSION: Focused ultrasound is not effective for non-invasive body contouring among Southern Asians as compared with Caucasian. Such observation is likely due to smaller body figures. Design modifications can overcome this problem and in doing so, improve clinical outcome. PMID- 20014262 TI - Cryolipolysis for subcutaneous fat layer reduction. AB - BACKGROUND AND OBJECTIVE: Cryolipolysis is a unique non-invasive method for the selective reduction of fat cells with controlled, localized cooling. It is important, therefore, to understand the potential efficacy and safety of this new procedure for fat layer reduction. MATERIALS AND METHODS: A review of the literature associated with cryolipolysis was performed to evaluate the findings from pre-clinical and clinical studies with respect to the mechanism of action, efficacy, and safety. RESULTS: Cryolipolysis has demonstrated efficacy in both human and animal studies. Histology findings also confirm the selective reduction of fat in both humans and animals, with evidence of a gradual thinning of the fat layer over a period of two to four months. Importantly, cryolipolysis has not produced any significant adverse side effects in studies to date and any noted effects have been minor and temporary. CONCLUSION: Although the mechanism of action for cryolipolysis is not yet completely understood, the efficacy and safety of this non-invasive procedure for fat layer reduction has been demonstrated in the studies available to data. Further studies willl assist in identifying the mechanism and elucidate the full potential of this technology to perform safe, non-invasive fat reduction for areas of local fat accumulation. PMID- 20014263 TI - The emerging preclinical and clinical studies regarding fat removal technologies, devices and procedures. Introduction. PMID- 20014264 TI - Two patients with EP300 mutations and facial dysmorphism different from the classic Rubinstein-Taybi syndrome. AB - Rubinstein-Taybi syndrome (RTS) is characterized by mental retardation, broad thumbs and great toes and a recognizable craniofacial phenotype. Causative mutations have been described in the CREBBP and EP300 genes. Here we present a 19 year-old woman and an unrelated 3-year-old boy, both with broad thumbs and halluces, but with facial aspects distinct from those of typical RTS. The woman had a marked learning disability, but no mental retardation. We identified a de novo c.7100delC mutation in EP300 (which predicts p.P2366RfsX35) in the woman and an apparently de novo c.638delG mutation in the boy, which predicts p.G213EfsX6. Mutations in EP300 are a known but rare cause of RTS. Only five other patients have been reported. We propose that individuals with EP300 mutations may exhibit a slightly different phenotype compared to individuals with CREBBP mutations, with milder cognitive impairment, more pronounced microcephaly, absent or mild downslanting of palpebral fissures, distinct arched eyebrows, and greater degree of retrognathia. PMID- 20014265 TI - Controlled volumetric heating of subcutaneous adipose tissue using a novel radiofrequency technology. AB - BACKGROUND AND OBJECTIVE: The objective of the present study was to demonstrate the feasibility of varying the size of the heating volume of subcutaneous adipose tissue using a novel radiofrequency (RF) technology that controls the delivered energy distribution on the skin surface. STUDY DESIGN/MATERIALS AND METHODS: Changes in the distribution of the electric potential at the skin surface due to frequency adjustment of a novel RF device were experimentally characterized on human skin at 500 kHz, 1, 2, 3, and 4 MHz. These measurements were used to model RF-induced electric fields and power absorption. Thermal measurements in ex vivo animal models were used to complement the initial mathematical modeling. RESULTS: At 500 kHz the electric potential on the skin surface was nearly constant across the RF applicator surface. At 4 MHz the electric potential dropped 30% from the center to the edge of the RF applicator. At the centerline of the RF applicator, modeling shows that within a 3 cm subcutaneous fat layer the absorbed power at the bottom layer was 40% less than that at the top for 500 kHz. The absorbed power decreased 80% for 4 MHz. Temperature measurements show uniform heating across a horizontal array of probes with 500 kHz. Temperatures were significantly higher at the center probes for 4 MHz. Cross-sectional radiometric temperature maps show a larger heated tissue cross-section using 500 kHz as opposed to 4 MHz. CONCLUSIONS: As the frequency increases (i) the electric potential at the skin surface decreases from the center to the edge of the RF applicator; (ii) the difference between the power absorbed at the top and bottom of the subcutaneous fat layer increases; (iii) the difference between the power absorbed at the center and the periphery of the exposed subcutaneous fat volume also increases; and, consequently, (iv) the size of the heated subcutaneous fat volume decreases. Such a device when used in humans may allow for differential delivery of heat to varying fat thicknesses and anatomic areas. PMID- 20014266 TI - Highly enantioselective recognition of structurally diverse alpha hydroxycarboxylic acids using a fluorescent sensor. PMID- 20014267 TI - Asymmetric counteranion-directed transition-metal catalysis: enantioselective epoxidation of alkenes with manganese(III) salen phosphate complexes. PMID- 20014269 TI - Synthesis of [8]cycloparaphenylene from a square-shaped tetranuclear platinum complex. PMID- 20014270 TI - In vitro recombination mediated by G-quadruplexes. PMID- 20014268 TI - Rhodium-catalyzed asymmetric intramolecular hydroamination of unactivated alkenes. PMID- 20014271 TI - Structure and binding of peptide-dendrimer ligands to vitamin B12. AB - The third-generation peptide-dendrimer B1 (AcES)8(BEA)4(K-Amb-Y)2BCD-NH2 (B=branching (S)-2,3-diaminopropanoic acid, K=branching lysine, Amb=4-aminomethyl benzoic acid) is the first synthetic model for cobalamin-binding proteins and binds cobalamin strongly (K(a)=5.0 x 10(6) M(-1)) and rapidly (k(2)=346 M(-1) s( 1)) by coordination of cobalt to the cysteine residue at the dendrimer core. A structure-activity relationship study is reported concerning the role of negative charges in binding. Substituting glutamates (E) for glutamines (Q) in the outer branches of B1 to form N3 (AcQS)8(BQA)4(B-Amb-Y)(2)BCD-NH2 leads to stronger (K(a)=12.0 x 10(6) M(-1)) but slower (k(2)=67 M(-1) s(-1)) cobalamin binding. CD and FTIR spectra show that the dendrimers and their cobalamin complexes exist as random-coil structures without aggregation in solution. The hydrodynamic radii of the dendrimers determined by diffusion NMR either remains constant or slightly decreases upon binding to cobalamin; this indicates the formation of compact, presumably hydrophobically collapsed complexes. PMID- 20014272 TI - Mechanistic insights into selectivity control for heterogeneous olefin oxidation: styrene oxidation on Au(111). AB - We demonstrate that intermolecular interactions, controlled by both oxygen and styrene coverage, alter reaction selectivity for styrene oxidation on oxygen covered Au(111). Several partial oxidation products are formed--styrene oxide, acetophenone, benzoic acid, benzeneacetic acid, and phenylketene--in competition with combustion. The maximum ratio of the yields of styrene oxide to the total CO(2) produced is obtained for the maximum styrene coverage for the first two layers (0.28 ML) adsorbed on Au(111) precovered with 0.2 ML of O. Furthermore, our reactivity and infrared studies support a mechanism whereby styrene oxidation proceeds via two oxametallacycle intermediates which, under oxygen-lean conditions, lead to the formation of styrene oxide, acetophenone, and phenylketene. Benzoate, identified on the basis of infrared reflection absorption spectroscopy, is converted into benzoic acid during temperature-programmed reaction. These results demonstrate the ability to tune the epoxidation selectivity using reactant coverages and provide important mechanistic insight into styrene oxidation reactions. PMID- 20014273 TI - Socially biased learning among adult cottontop tamarins (Saguinus oedipus). AB - We presented adult cottontop tamarins (Saguinus oedipus) with a novel foraging task that had been used previously to examine socially biased learning of juvenile observers [Humle & Snowdon, Animal Behaviour 75:267-277, 2008]. The task could be solved in one of two ways, and thus allowed for an analysis of behavioral matching between an observer and a skilled demonstrator (trained to use one of the two methods exclusively). Because the demonstrator was an adult in both this study and the juvenile study, the influence of the observer's age could be isolated and examined, as well as the behavior of demonstrators toward observers of different ages. Our main goals were to (1) compare adults and juveniles acquiring the same task to identify how the age of the observer affects socially biased learning and (2) examine the relationship between socially biased learning and behavioral matching in adults. Although adults spent less time observing the trained demonstrators than did juveniles, the adults were more proficient at solving the task. Furthermore, even though observers did not overtly match the behavior of the demonstrator, observation remained an important factor in the success of these individuals. The findings suggested that adult observers could extract information needed to solve a novel foraging task without explicitly matching the behavior of the demonstrator. Adult observers begged much less than juveniles and demonstrators did not respond to begging from adult. Skill acquisition and the process of socially biased learning are, therefore, age dependent and are influenced by the behavioral interactions between observer and demonstrator. To what extent this holds true for other primates or animal species still needs to be more fully investigated and considered when designing experiments and interpreting results. PMID- 20014274 TI - Gastrointestinal parasites of the chimpanzee population introduced onto Rubondo Island National Park, Tanzania. AB - The release of any species into a novel environment can evoke transmission of parasites that do not normally parasitize the host as well as potentially introducing new parasites into the environment. Species introductions potentially incur such risks, yet little is currently known about the parasite fauna of introduced primate species over the long term. We describe the results of long term monitoring of the intestinal parasite fauna of an unprovisioned, reproducing population of chimpanzees introduced 40 years earlier (1966-1969) onto Rubondo Island in Lake Victoria, Tanzania, a non-native habitat for chimpanzees. Two parasitological surveys (March 1997-October 1998 and October 2002-December 2005) identified Entamoeba spp. including E. coli, Iodamoeba buetschlii, Troglodytella abrassarti, Chilomastix mesnili, Trichuris sp., Anatrichosoma sp., Strongyloides spp., Strongylida fam. gen. sp., Enterobius anthropopitheci, Subulura sp., Ascarididae gen. sp., and Protospirura muricola. The parasite fauna of the Rubondo chimpanzees is similar to wild chimpanzees living in their natural habitats, but Rubondo chimpanzees have a lower prevalence of strongylids (9%, 3.8%) and a higher prevalence of E. anthropopitheci (8.6%, 17.9%) than reported elsewhere. Species prevalence was similar between our two surveys, with the exception of Strongyloides spp. being higher in the first survey. None of these species are considered to pose a serious health risk to chimpanzees, but continued monitoring of the population and surveys of the parasitic fauna of the two coinhabitant primate species and other animals, natural reservoir hosts of some of the same parasites, is important to better understand the dynamics of host-parasite ecology and potential long-term implications for chimpanzees introduced into a new habitat. PMID- 20014275 TI - Ammonia triggers exocytotic release of L-glutamate from cultured rat astrocytes. AB - Ammonia toxicity to the brain involves NMDA receptor overactivation and glutamate excitotoxicity. The mechanisms underlying glutamate release from astrocytes in response to ammonia were addressed in this study. In cultured rat astrocytes, glutamate immunoreactivity (IR) was punctate and partly colocalized with transfected VAMP2-YFP. NH(4)Cl (5 mmol/L) and hypoosmotic exposure (205 mosmol/L) induced a rapid colchicine-sensitive loss of cellular glutamate and glutamate appearance in the extracellular space. The NH(4)Cl-induced glutamate loss from astrocytes was strongly blunted after transfection of the cells with VAMP2 siRNA. Ammonia-induced exocytosis of VAMP2-YFP expressing vesicles was shown by total internal reflection fluorescence microscopy (TIRF-M). Glutamate exocytosis in response to ammonia was sensitive to chelation of Ca(2+), cyclooxygenase inhibition by indomethacin and colchicine. Ammonia triggered the rapid formation of prostanoids, which were identified as upstream events in ammonia-induced glutamate exocytosis. Also, addition of prostaglandin E(2) or of tumor necrosis factor (TNF)-alpha triggered glutamate exocytosis. Inhibition of ammonia-induced glutamate exocytosis after transfection of VAMP2 siRNA inhibited ammonia-induced RNA oxidation. It is concluded that ammonia triggers a prostanoid- and Ca(2+) dependent glutamate exocytosis, which is essential for induction of ammonia induced RNA oxidation. PMID- 20014276 TI - Gelsolin is required for macrophage recruitment during remyelination of the peripheral nervous system. AB - Reorganization of the actin cytoskeleton is necessary for Schwann cell proliferation, migration and for the morphological changes associated with sorting, ensheathing and myelination of axons. Such reorganization requires regulated severing and depolymerization of actin filaments. Gelsolin is an actin filament severing protein expressed in many cell types including Schwann cells. Using Gelsolin knockout mice, we investigated the role of this protein in the myelination and remyelination of the peripheral nervous system. Our results show that although gelsolin is not necessary for developmental myelination, it is required for timely remyelination of the sciatic nerve following crush injury. Gelsolin is necessary for macrophage motility in culture, and its absence is likely to impair the recruitment of macrophages to the injury site. PMID- 20014277 TI - Blueberry supplementation attenuates microglial activation in hippocampal intraocular grafts to aged hosts. AB - Transplantation of central nervous tissue has been proposed as a therapeutic intervention for age-related neurodegenerative diseases and stroke. However, survival of embryonic neuronal cells is hampered by detrimental factors in the aged host brain such as circulating inflammatory cytokines and oxidative stress. We have previously found that supplementation with 2% blueberry in the diet increases graft growth and neuronal survival in intraocular hippocampal grafts to aged hosts. In the present study we explored possible biochemical mechanisms for this increased survival, and we here report decreased microglial activation and astrogliosis in intraocular hippocampal grafts to middle-aged hosts fed a 2% blueberry diet. Markers for astrocytes and for activated microglial cells were both decreased long-term after grafting to blueberry-treated hosts compared with age-matched rats on a control diet. Similar findings were obtained in the host brain, with a reduction in OX-6 immunoreactive microglial cells in the hippocampus of those recipients treated with blueberry. In addition, immunoreactivity for the pro-inflammatory cytokine IL-6 was found to be significantly attenuated in intraocular grafts by the 2% blueberry diet. These studies demonstrate direct effects of blueberry upon microglial activation both during isolated conditions and in the aged host brain and suggest that this nutraceutical can attenuate age-induced inflammation. PMID- 20014278 TI - Oct4 and klf4 reprogram dermal papilla cells into induced pluripotent stem cells. AB - Direct reprogramming of somatic cells into induced pluripotent stem (iPS) cells by only four transcription factors (Oct4, Sox2, Klf4, and c-Myc) has great potential for tissue-specific regenerative therapies, eliminating the ethical issues surrounding the use of embryonic stem cells and the rejection problems of using non-autologous cells. The reprogramming efficiency generally is very low, however, and the problems surrounding the introduction of viral genetic material are only partially investigated. Recent efforts to reduce the number of virally expressed transcription factors succeeded at reprogramming neural stem cells into iPS cells by overexpressing Oct4 alone. However, the relative inaccessibility and difficulty of obtaining neural cells in humans remains to be resolved. Here we report that dermal papilla (DP) cells, which are specialized skin fibroblasts thought to instruct hair follicle stem cells, endogenously express high levels of Sox2 and c-Myc, and that these cells can be reprogrammed into iPS cells with only Oct4 and Klf4. Moreover, we show that DP cells are reprogrammed more efficiently than skin and embryonic fibroblasts. iPS cells derived from DP cells expressed pluripotency genes and differentiated into cells from all germ layers in vitro and widely contributed to chimeric mice in vivo, including the germline. Our work establishes DP cells as an easily accessible source to generate iPS cells with efficiency and with less genetic material. This opens up the possibility of streamlined generation of skin-derived, patient-specific pluripotent stem cells and of ultimately replacing the remaining two factors with small molecules for safe generation of transplantable cells. PMID- 20014280 TI - Analysis of nifedipine absorption from soft gelatin capsules using PBPK modeling and biorelevant dissolution testing. AB - Delayed absorption of nifedipine when administered as a 20 mg immediate release soft gelatin capsule to fasted volunteers has been reported. Physiologically based pharmacokinetic (PBPK) modeling and in vitro dissolution data were used to explore our hypothesis that at high doses of nifedipine it precipitates in the stomach. Plasma concentration-time profiles following different doses of nifedipine were simulated using commercial PBPK software and compared to in vivo data. In vitro dissolution tests were performed with Adalat 10 mg capsules in different volumes of fasted state simulated gastric fluid (FaSSGF). The discrepancy in plasma concentration-time profiles between the different nifedipine doses could be well simulated, assuming protracted dissolution for the 20 mg dose. Nifedipine release from one Adalat 10 capsule in 250 or 500 mL FaSSGF was completed within 15 min whereas when release from two capsules, corresponding to 20 mg nifedipine, was studied in 250 mL FaSSGF, a maximum of about 75% drug dissolved was observed after 15 min followed by a decline in the % dissolved to a final value of approximately 40%. Based on the in silico and in vitro results it can be concluded that the observed prolongation in nifedipine absorption following the 20 mg dose was likely caused by nifedipine precipitation in human stomach. PMID- 20014279 TI - NF kappaB activation in embryonic endothelial progenitor cells enhances neovascularization via PSGL-1 mediated recruitment: novel role for LL37. AB - Embryonal endothelial progenitor cells (eEPCs) are capable of inducing therapeutic angiogenesis in a chronic hind limb model. However, the proportion of eEPCs recruited to the ischemic tissue appears to be a limiting step for the induction of cell-based therapeutic neovascularization. In the present study, we primed eEPCs with the human cathelicidin LL37 (hCAP-18) ex vivo to selectively enhance the eEPC-dependent gain of perfusion in vivo and elucidated the mechanism of action of LL37 on eEPCs. Seven days after femoral artery excision, 5 x 10(6) eEPCs (wt, wild type; p65t, transiently p65 transient; p65s, stable p65 transfected; LL37-eEPCs, LL37 peptide preincubated) were retroinfused into the anterior tibial vein. Recruitment of diI-labeled eEPCs in the ischemic gastrocnemic muscle was investigated 2 days later, whereas collateral growth and perfusion score (obtained by fluorescent microspheres) were assessed at day 7 and day 35 and are given as percentage of day 7 level. Capillary/muscle fiber ratio in the ischemic lower limb was obtained at day 35. Embryonic EPC recruitment in vitro and in vivo was found elevated after LL37 and p65t pretreatment, but not in p65s-eEPCs displaying increased IkappaBalpha or after LL37 in IkappaB-DN overexpressing eEPCs. Using LL37- and p65t-eEPCs, collateral growth (181 +/- 10% and 165 +/- 8%, respectively) surpassed that of wt-eEPCs (135 +/- 7%), increasing perfusion ratio (208 +/- 20% and 210 +/- 17% vs. 142 +/- 12% in wt-eEPCs, respectively), whereas p65s-eEPCs exerted no additive effect (collateral growth 130 +/- 8%; perfusion ratio 155 +/- 15%). Moreover, p65t-eEPC-induced neovascularization was abrogated by blocking antibodies against E-selectin and P selectin glycoprotein ligand-1 (PSGL-1). We conclude that NF kappaB activation by LL37 or transient p65-transfection increases functionally relevant eEPC recruitment to ischemic muscle tissue via induction of PSGL-1 and E-selectin. PMID- 20014281 TI - Enhanced skin permeation of naltrexone by pulsed electromagnetic fields in human skin in vitro. AB - The aim of the present study was to evaluate the skin permeation of naltrexone (NTX) under the influence of a pulsed electromagnetic field (PEMF). The permeation of NTX across human epidermis and a silicone membrane in vitro was monitored during and after application of the PEMF and compared to passive application. Enhancement ratios of NTX human epidermis permeation by PEMF over passive diffusion, calculated based on the AUC of cumulative NTX permeation to the receptor compartment verses time for 0-4 h, 4-8 h, and over the entire experiment (0-8 h) were 6.52, 5.25, and 5.66, respectively. Observation of the curve indicated an initial enhancement of NTX permeation compared to passive delivery whilst the PEMF was active (0-4 h). This was followed by a secondary phase after termination of PEMF energy (4-8 h) in which there was a steady increase in NTX permeation. No significant enhancement of NTX penetration across silicone membrane occurred with PEMF application in comparison to passively applied NTX. In a preliminary experiment PEMF enhanced the penetration of 10 nm gold nanoparticles through the stratum corneum as visualized by multiphoton microscopy. This suggests that the channels through which the nanoparticles move must be larger than the 10 nm diameter of these rigid particles. PMID- 20014282 TI - Leukemia inhibitory factor promotes Hsp90 association with STAT3 in mouse embryonic stem cells. AB - Self-renewal of in vitro cultured mouse embryonic stem (mES) cells is dependent on the presence of leukemia inhibitory factor (LIF). LIF induces overexpression and tyrosine phosphorylation of STAT3 (signal transducer and activator of transcription 3) and its subsequent nuclear translocation. The molecular chaperone heat shock protein 90 (Hsp90) is involved in the activation and maturation of a wide variety of substrate proteins. We investigated the effect of LIF withdrawal on the protein expression levels of STAT3 and Hsp90 and on the interactions between STAT3 and Hsp90. Taken together the data presented here suggest that LIF promotes the interaction of Hsp90 with STAT3 during self renewal, indicating a potentially pivotal role for Hsp90 in the LIF-based maintenance of self-renewal of mouse embryonic stem cells. PMID- 20014283 TI - Poly(DL:lactic acid-castor oil) 3:7-bupivacaine formulation: reducing burst effect prolongs efficacy in vivo. AB - Prolonged analgesia may be achieved using a single injection of slow-release local anesthetic formulation. The study objective was to improve the efficacy of a previously reported formulation comprising 10% bupivacaine in poly(DL:lactic acid co castor oil) 3:7. The polymer was loaded with 15% bupivacaine and injected through a 22G needle close to the sciatic nerve of ICR mice. Sensory and motor nerve blockade were measured. The efficacy and toxicity of the polymer-drug combination were determined. Sixty percent of the incorporated bupivacaine was released during 1 week in vitro. During in vitro release no burst effect was seen, suggesting low toxicity of the formulation. Single injection of 0.1 mL of 15% polymer-bupivacaine formulation caused motor block that lasted 64 h and sensory block that lasted 96 h. The MTD of the polymer-drug formulation was established as 0.175 mL. Microscopic examination of the injection sites revealed reversible nerve inflammation and normal internal organs. The polymer poly(DL:lactic acid co castor oil) 3:7 is a safe carrier for prolonged activity of bupivacaine up to 96 h. The increase of drug load in the formulation reduces the drug release rates due to stronger polymer-drug interactions and higher overall hydrophobicity of the formulation. PMID- 20014284 TI - Micro-CT analysis of matrix-type drug delivery devices and correlation with protein release behaviour. AB - A series of matrix-type drug delivery devices comprising a continuous phase of microporous poly(epsilon-caprolactone) (PCL) and a dispersed phase of protein particles (gelatin) with defined size ranges (45-90, 90-125 and 125-250 microm) were produced by rapidly cooling suspensions in dry ice followed by solvent extraction from the hardened material. High protein loadings (38-44%, w/w) were achieved and highly efficient protein release (90% of the initial load) was obtained over time periods of 3-11 days depending on particle loading and size range. The duration of protein release was extended from 3 to 11 days by reducing the protein load. Quantitative analysis of Micro-CT images identified a three to four times increase in the population of sub-40 microm pores in those matrices which gave rise to accelerated protein release in 24 h (40% rising to 80%) and reduced duration of protein release (11-3 days). Formation of a high density of channels and fissures (connects) between the particles is indicated, which facilitate fluid ingress and diffusion of solubilised protein molecules. Micro-CT analysis also confirmed the uniformity of particle distribution in the matrices and provided measurements of macroporosity within 5-30% of the theoretical value for materials displaying irregular shaped macropores larger than 90 microm. These findings demonstrate the utility of Micro-CT for optimising the formulation and performance of matrix-type delivery devices for macromolecular entities. PMID- 20014285 TI - Effect of encapsulating arginine containing molecules on PLGA: a solid-state NMR study. AB - Design of polymer-drug composites based on the lactide/glycolic acid often rely on the chemical complementarity between the polymer and functional groups in a pharmaceutical guest. We previously characterised decapeptide (AZD)/poly(D,L lactide-co-glycolide) (PLGA) film formulations aiming at localising the interacting groups responsible for the changes in the bulk properties of the polymer matrix and understanding the mechanism of stabilisation of the drug into the polymer matrix. The results suggested interactions to occur between the arginine residue in the peptide and the carbonyl end group of the polymer chains. In order to clarify the role of arginine in directing the drug-polymer interactions, arginine and hexapeptide containing arginine were encapsulated in a PLGA 50/50 polymer. Variable temperature T1 rh H measurements and WISE experiments indicated significant changes in the local dynamics of the polymer chains. These effects were enhanced near and above T(g) suggesting the presence of guests promote the appearance of backbone motion of the polymer chains. The localisation of the interactions on the carbonyl groups of the polymer was further confirmed by the WISE experiments. PMID- 20014286 TI - Effects of nutritional supplements on aggression, rule-breaking, and psychopathology among young adult prisoners. AB - In an earlier study, improvement of dietary status with food supplements led to a reduction in antisocial behavior among prisoners. Based on these earlier findings, a study of the effects of food supplements on aggression, rule breaking, and psychopathology was conducted among young Dutch prisoners. Two hundred and twenty-one young adult prisoners (mean age=21.0, range 18-25 years) received nutritional supplements containing vitamins, minerals, and essential fatty acids or placebos, over a period of 1-3 months. As in the earlier (British) study, reported incidents were significantly reduced (P=.017, one-tailed) in the active condition (n=115), as compared with placebo (n=106). Other assessments, however, revealed no significant reductions in aggressiveness or psychiatric symptoms. As the incidents reported concerned aggressive and rule-breaking behavior as observed by the prison staff, the results are considered to be promising. However, as no significant improvements were found in a number of other (self-reported) outcome measures, the results should be interpreted with caution. PMID- 20014287 TI - Alveolar ridge augmentation in irradiated rabbit mandibles. AB - Oral carcinomas are frequently treated with a nonsegmental mandibulectomy plus radiotherapy. Improving the quality of life of these patients depends on the possibilities for dental rehabilitation. The aim of this study was to increase the alveolar ridge height. Twelve white New Zealand rabbits underwent surgery to produce a 11 x 9 x 7 mm(3) horizontal bicortical full-thickness defect at the alveolar ridge of the left mandible. Six were implanted with a composite associating resorbable collagen membrane filled with micro-macroporous biphasic calcium phosphate granules. After a daily radiation delivery schedule for 4 weeks, a total autologous bone marrow graft was injected percutaneously into the center of the implant. All animals were sacrificed at 16 weeks. Successful osseous colonization was observed in all implants. Significant ridge augmentation was observed (p = 0.0349) in the implanted group compared with the control group. This study contributed to producing an experimental model for oncological mandible defects in rabbits. PMID- 20014288 TI - Growth factor delivery through electrospun nanofibers in scaffolds for tissue engineering applications. AB - Tissue engineering scaffolds should ideally mimic the natural ECM in structure and function. Electrospun nanofibrous scaffolds are easily fabricated and possess a biomimetic nanostructure. Scaffolds can mimic ECM function by acting as a depot for sustained release of growth factors. bFGF, an important growth factor involved in tissue repair and mesenchymal stem cell proliferation and differentiation, is a suitable candidate for sustained delivery from scaffolds. In this study, we present two types of PLGA nanofibers incorporated with bFGF, fabricated using the facile technique of blending and electrospinning (Group I) and by the more complex technique of coaxial electrospinning (Group II). bFGF was randomly dispersed in Group I and distributed as a central core within Group II nanofibers; both scaffolds showed similar protein encapsulation efficiency and release over 1-2 weeks. Although both scaffold groups favored bone marrow stem cell attachment and subsequent proliferation, cells cultured on Group I scaffolds demonstrated increased collagen production and upregulated gene expression of specific ECM proteins, indicating fibroblastic differentiation. The study shows that the electrospinning technique could be used to prolong growth factor release from scaffolds and an appropriately sustained growth factor release profile in combination with a nanofibrous substrate could positively influence stem cell behavior and fate. PMID- 20014289 TI - In vitro uptake of amphiphilic, hydrogel nanoparticles by J774A.1 cells. AB - We here report improved synthesis and in vitro interactions of amphiphilic hydrogel nanoparticles with the macrophage cell line J774A.1. Nanoparticles comprising dispersed hydrophobic nanodomains of poly(propylene glycol) within a continuous phase of hydrophilic poly (ethylene glycol) (PEG) were prepared via inverse emulsion crosslinking polymerization, using acrylated PEG and Pluronic F127 as macromonomer blocks. Functionality and fluorescent labeling were achieved through incorporation of reactive comonomers and a posteriori reaction with fluorescein, respectively. When introduced to a static cell culture of adhered J774A.1 macrophages, the cells internalized these hydrogel nanoparticles in a dose- and time- dependent manner through clathrin-mediated and other pathways. Amphiphilic nanoparticle uptake was however dramatically lower than that of a model system (Fluospheres) and similar to PEG-coated colloids reported in the literature, which are considered "stealth." Our findings support the potential of the nanoparticles presented here as long-circulating drug carriers. PMID- 20014290 TI - Influence of anodization on the adhesion of calcium phosphate coatings on titanium substrates. AB - Electrochemical deposition is an attractive technique for the deposition of calcium phosphate, especially hydroxyapatite, on titanium implants. However, the adhesion of these coatings to the titanium substrates needs to be improved for clinical use. It is demonstrated that anodization of a titanium alloy does marginally increase the adhesion of calcium phosphate coatings. Although scratch test measurements on coatings deposited at a constant potential appear to suggest that adhesion improves with increased thickness of the anodized layer, when a constant current is used to deposit the coatings their adhesion becomes independent of the thickness of the anodized layer. This apparent contradiction is explained by the thicker oxides acting as larger series resistors that reduce the magnitude of the current density when deposition is conducted at a constant potential. The resulting lower current density is responsible for increased adhesion of the calcium phosphate coating. It was also observed that surface roughness affects the interfacial adhesion strength between the coating and the titanium substrate, with a more adherent coating being formed over a rough surface. However, adhesion becomes independent of surface finish at levels smoother than 600 grit, suggesting that mechanical interlocking is not the sole force at play. PMID- 20014291 TI - Poly(vinyl alcohol)/halloysite nanotubes bionanocomposite films: Properties and in vitro osteoblasts and fibroblasts response. AB - In this study, transparent poly(vinyl alcohol) (PVA) and PVA/halloysite nanotubes (HNTs) bionanocomposite films were prepared by solution casting and glutaraldehyde (GA) crosslinking. The surface topography and chemistry of the films were characterized by atomic force microscopy (AFM) and attenuated total reflection Fourier transform infrared (ATR-FTIR) spectroscopy, respectively. Blending with HNTs induced changes in nanotopography and surface chemistry of PVA films. The mechanical properties of PVA were enhanced by the incorporated HNTs. The stain-induced crystallization was confirmed by DSC after tensile test. MC3T3 E1 osteoblast-like and NIH 3T3 fibroblast cells were cultured on neat PVA and PVA/HNTs films to evaluate the effects of surface nanotopography and composition on cell behavior. The observations indicated that MC3T3-E1 cell behavior strongly responded to surface nanotopography. On nanotube-dominant surface, cells exhibited a significantly higher level of adhesion than on neat PVA film, whereas neat PVA showed higher degree of osteoblast proliferation compared with PVA/HNTs. In vitro fibroblasts response demonstrated that both neat PVA and PVA/HNTs nanocomposite films were biocompatible and PVA/HNTs films favored to fibroblasts attach and growth below 7.5 wt % of HNTs incorporated. In summary, these results provided insights into understanding of PVA and PVA/HNTs bionanocomposite films in potential applications in bone tissue engineering and drug delivery systems. PMID- 20014292 TI - Transforming growth factor-beta activated kinase 1 signaling pathways regulate TNF-alpha production by titanium alloy particles in RAW 264.7 cells. AB - Implant particles may induce inflammatory response by activating the nuclear factor kappaB (NFkappaB) and mitogen activated protein kinases (MAPK). Previous studies have shown that these signaling pathways are involved in the transforming growth factor-beta activated kinase 1 (TAK1) in signaling cascades induced by the receptor activator of NFkappaB ligand (RANKL) and tumor necrosis factor-alpha (TNF-alpha) as well as interleukin-1beta (IL-1beta). In this study, the roles of the TAK1 pathway in the production of the pro-inflammatory cytokine TNF-alpha in RAW 264.7 cells exposed to titanium alloy particles were investigated. Endogenous TAK1 was phosphorylated upon simulating RAW 264.7 cells by titanium alloy particles. The critical role for TAK1 in p38MAPK and NFkappaB activation was as well confirmed by the inhibition of p38MAPK and NFkappaB activity following 5Z-7 oxozeaenol, a selective inhibitor of TAK1. Furthermore, it was found that TNF alpha was completely blocked by 5Z-7-oxozeaenol in RAW 264.7 cells. These results suggest that the TAK1-MAPK-NFkappaB signaling pathways may be a potential pharmacological target that can prevent instability for arthroplasty prosthesis. PMID- 20014293 TI - Titanium is not "the most biocompatible metal" under cathodic potential: The relationship between voltage and MC3T3 preosteoblast behavior on electrically polarized cpTi surfaces. AB - An electrochemically controlled system has been developed which allows for cell culture directly on electrically polarized metal surfaces with simultaneous control and assessment of the electrochemical current, potential, and impedance of the interface. This system was utilized in this study to assess the interactions between electrochemically polarized commercially pure titanium (cpTi) and MC3T3 preosteoblast cells. Cells were cultured on CpTi for 24 h at static potentials between -1000 mV and +1000 mV vs. Ag/AgCl and cell morphology (SEM and cell area) and viability (MTT and Live-Dead assay) were assessed along with the electrochemical current densities and surface oxide impedance properties. The results indicate that cathodic polarization in the range of -600 mV to -1000 mV markedly reduces the spreading and viability of cells cultured directly on cpTi within 24 h, while anodic polarization (-300 mV to +1000 mV) out to 72 h shows no difference in cell behavior as compared to the OCP condition. Analysis of the relationship between the cell outcomes and the electrochemical current densities and impedance indicated the presence of voltage-dependent electrochemical thresholds (cathodic current density, i(c) > 1.0 microA/cm(2), R(p) < 10(5) Omega cm(2)) which may control the biocompatibility of cpTi. In addition, these outcomes have direct clinical significance for modular orthopedic implants whose potential can shift, via fretting corrosion, down into the range of potentials exhibiting poor cell behavior. PMID- 20014294 TI - In vivo evaluation of acellular human dermis for abdominal wall repair. AB - Limitations of synthetic biomaterials for abdominal wall repair have led investigators to seek naturally derived matrices, such as human acellular dermis, because of their excellent biocompatibility and their ability to naturally interface with host tissues with minimal tissue response. In this study, we investigated two different biomaterials derived from human dermis (FlexHD acellular dermis and FlexHD acellular dermis-thick) in a rabbit abdominal hernia repair model. One quarter of the abdominal wall was replaced with each biomaterial, and the animals were followed for up to 24 weeks. Rabbit hernias repaired with AlloDerm, a commercially available acellular dermal matrix, and sham operated animals served as controls. Retrieved samples of these implants were assessed grossly and histologically. Collagen production measurements and tension studies were performed. FlexHD acellular dermis, FlexHD acellular dermis thick, and AlloDerm maintained their strength in the rabbit hernia repair model with no incidence of hernia formation or bowel adhesion. The exact size measurements at 24 weeks were 217.0 +/- 20.9% for FlexHD acellular dermis, 200.8 +/- 23.5% for FlexHD acellular dermis-thick, and 209.7 +/- 32.9% for AlloDerm. Macroscopic and microscopic evaluation showed excellent integration and tissue formation. All biomaterials studied harbored cells that produced new collagen fibers, and a six-fold increase in these fibers was observed at 24 weeks. This study shows that acellular biomaterials derived from human dermis are suitable for abdominal hernia repair. PMID- 20014295 TI - Reconstruction of a mandibular defect with autogenous, autoclaved bone grafts and tissue engineering: An in vivo pilot study. AB - Reconstruction of bone defects with autogenous, autoclaved bone grafts has already been described but does have one major insuperable problem-the loss of the ostoinductive potential of the graft. In this study, we investigated if autogenous, autoclaved grafts in combination with tissue engineered bone can overcome this problem. An en-bloc resection was done in the mandible of eight pigs. The grafts were autoclaved and filled with autogenous, osseogen differentiated bone marrow cells and compared with four animals without bone marrow cells. After 120 days, the specimens were qualitatively and quantitatively evaluated by means of microradiography and light microscopy. Within the experimental group, osseous remodeling was detected in all cases and new bone formation was visible. Quantitative assessment of the osseous bridging of the osteotomy sites was significantly higher in the test group in comparison with the control group (p = 0.03). The histological evaluation by means of an osseous integration of the grafts revealed a statistically significant difference between both groups as well (p = 0.01). The results of this study indicate that the method investigated hereby represents a further possibility in the therapy of bony defects, such as those arising as a result of tumor operations. PMID- 20014296 TI - Macrophage and osteoblast responses to biphasic calcium phosphate microparticles. AB - The aim of this work was to investigate in vitro the biological events leading to ectopic bone formation in contact with microporous biphasic calcium phosphate (BCP) ceramics. After implantation, microparticles may arise from their degradation and induce an inflammatory response involving macrophages. The secretion of pro-inflammatory cytokines may affect the differentiation of osteoblasts. Mouse macrophage-like (J774) and osteoblast-like (MC3T3-E1) cells were cultured in the presence of BCP microparticles of different sizes (<20, 40 80, or 80-200 microm). The smallest microparticles decreased the viability of both cell types as measured with LDH and methyl tetrazolium salt assays, and enhanced the secretion of pro-inflammatory cytokines (IL-6 and TNF-alpha) by macrophages after 24 h, as revealed by ELISA. Osteoblastic cells were then cultured for 96 h in the presence of these pro-inflammatory cytokines and their differentiation studied by RT-PCR. MC3T3-E1 cells cultured with TNF-alpha showed a decrease in osterix, PTH receptor (PTHR1), and osteocalcin gene expression. On the contrary, IL-6 enhanced the expression of osterix, Runx2, alkaline phosphatase, and osteocalcin compared with plastic. In conclusion, this study shows that the inflammatory response initiated by BCP microparticles may have both detrimental and beneficial effects on osteogenesis. PMID- 20014297 TI - Biocompatibility of porous polyethylene implants tissue-engineered by extracellular matrix and VEGF. AB - Rapid ingrowth of blood vessels and low inflammatory response are considered major prerequisites for successful implantation of biomaterials in reconstructive surgery. Aim of the present study was to evaluate whether tissue-engineered porous polyethylene (PPE) implants providing extracellular matrix components (ECM) and vascular endothelial growth factor (VEGF) in vivo improve microvascular ingrowth and mechanical integration with regard to initial inflammatory responses. PPE implants (3 x 3 x 0.1 mm(3), pore size approximately 100-200 microm) were tissue-engineered by incorporation of ECM components (GFR-Matrigel) adding recombinant murine VEGF (1 microg/mL) and grafted into dorsal skinfold chamber preparations of C57BL/6 mice. Control animals received uncoated implants or implants coated with ECM components alone (n = 6 per group). Using in vivo fluorescence microscopy angiogenic activity and inflammatory leukocyte endothelial cell interactions were analyzed for 2weeks. Finally, mechanical integration was quantified by measurement of dynamic desintegration strengths at the host-implant border. Functional vessel density, red blood cell velocity, and vessel diameters increased continuously in all groups indicating that rapid microvascular integration of PPE occurred even without incorporation of ECM or VEGF. However, a transient initial inflammatory response with increased leukocyte endothelial cell adherence on day 7 in uncoated control implants was efficiently reduced by incorporation of ECM and VEGF. Measurement of dynamic breaking strengths revealed no significant differences between the groups although there was a tendency to improved mechanical integration in tissue-engineered implants. Therefore, novel tissue- engineered constructs of PPE implants providing ECM and VEGF in high local concentrations can increase biocompatibility especially under unfavorable conditions for implantation. PMID- 20014298 TI - Micro and nano-scale in vitro 3D culture system for cardiac stem cells. AB - Despite the success to prevent or limit cardiovascular diseases, the restoration of the function of a damaged heart remains a formidable challenge. Cardiac stem cells (CSCs), with the capacity to differentiate into cardiomyocytes, hold great potential as a source of cells for regenerative medicine. A major challenge facing the clinical application of differentiated CSCs, however, is theability to generate sufficient numbers of cells with the desired phenotype. We previously established cell lines of CSCs using a c-kit antibody from adult rat hearts for use in regenerative medicine. C-kit -positive cardiac cells are well recognized as CSCs and have the potential to differentiate into cardiomyocytes. Here, before implant these cells in vivo, we first developed three-dimensional culture system (3D) using micro- and nano-scaled material. Sheets of poly(glycolic acid) (PGA) were fabricated by electrospinning. Composites of collagen-PGA were prepared that contained 0, 1.5, 3 or 6 mg of electrospun PGA nanofibers. The nanofibers were added as a sheet that formed a layer within the collagen sponge. The sponges were freeze-dried and then dehydrothermally crosslinked. A scanning electron microscopy (SEM)-based analysis of the surface of the sponges demonstrated a uniform collagenous structure regardless of the amount of PGA nanofibres included. The PGA nanofibers significantly enhanced the compressive strength of the collagen sponge. More CSCs attached to the collagen sponge incorporating 6 mg of PGA nanofibers than the sponge without PGA nanofibers. The attachment and proliferation of CSCs in the 3D culture was enhanced by incubation in a bioreactor perfusion system compared with 3D static and two-dimensional (2D; i.e. tissue culture plates) culture systems. The use of micro- and nano-scale materials in the fabrication of composites together with a 3D culture system is a very promising way to promote the culture of stem cells. (c) 2009 Wiley Periodicals, Inc. J Biomed Mater Res, 2010. PMID- 20014299 TI - QM/MM study of the absorption spectra of DsRed.M1 chromophores. AB - We report geometries and vertical excitation energies for the red and green chromophores of the DsRed.M1 protein in the gas phase and in the solvated protein environment. Geometries are optimized using density functional theory (DFT, B3LYP functional) for the isolated chromophores and combined quantum mechanical/molecular mechanical (QM/MM) methods for the protein (B3LYP/MM). Vertical excitation energies are computed using DFT/MRCI, OM2/MRCI, and TDDFT as QM methods. In the case of the red chromophore, there is a general blue shift in the excitation energies when going from the isolated chromophore to the protein, which is caused both by structural changes and by electrostatic interactions with the environment. For the lowest pipi* transition, these two factors contribute to a similar extent to the overall DFT/MRCI shift of 0.4 eV. An enlargement of the QM region to include active-site residues does not change the DFT/MRCI excitation energies much. The DFT/MRCI results are closest to experiment for both chromophores. OM2/MRCI and TDDFT overestimate the first vertical excitation energy by 0.3-0.5 and 0.2-0.4 eV, respectively, relative to the experimental or DFT/MRCI values. The experimental gap of 0.35 eV between the lowest pipi* excitation energies of the red (cis-acylimine) and green (trans-peptide) forms is well reproduced by DFT/MRCI and TDDFT (0.32 and 0.37 eV, respectively). A histogram spectrum for an equal mixture of the two forms, generated by OM2/MRCI calculations on 450 snapshots along molecular dynamics trajectories, matches the experimental spectrum quite well, with a gap of 0.23 eV and an overall blue shift of about 0.3 eV. DFT/MRCI appears as an attractive choice for calculating excitation energies in fluorescent proteins, without the shortcomings of TDDFT and computationally more affordable than CASSCF-based approaches. PMID- 20014300 TI - Use of interactive sessions and e-learning in teaching anatomy to first-year optometry students. AB - Students enrolled in the Optometry program at the University of Manchester are required to take a functional anatomy course during the first year of their studies. Low mean scores in the written examination of this unit for the past two academic years energized staff to rethink the teaching format. Interactive sessions lasting 20 minutes each were introduced during the two hour lecture sessions. In these sessions students reinforced their anatomical knowledge learned in lectures, through playing games such as anatomy bingo and solving anatomical anagrams. In addition, five e-learning modules were also introduced for students to complete in their own time. A pre- and postcourse questionnaire were distributed to obtain student views on their expectations of the course and interactive sessions. Comparisons were made between written examination results from 2008 to 2009 to written examination results from the previous five academic years to see if the interactive sessions and e-learning modules had any impact on student knowledge. In addition, comparisons were made between student performances on the functional anatomy course with their performance in all of the other assessments taken by the students during their first year of study. Analysis of the questionnaires showed that student's expectations of the course were fulfilled and the interactive sessions were well received by the majority. There was a significant increase (P < or = 0.01) in the mean examination score in 2008-2009 after introduction of the interactive sessions and e-learning modules compared with scores in previous years. The introduction of interactive sessions has increased student enjoyment of the module and along with the e-learning modules have had a positive impact on student examination results. PMID- 20014301 TI - Fluorescent high-content imaging allows the discrimination and quantitation of E LDL-induced lipid droplets and Ox-LDL-generated phospholipidosis in human macrophages. AB - Macrophage foam cells formed during uptake of atherogenic lipoproteins are a hallmark of atherosclerotic lesion development. In this study, human macrophages were incubated with two prototypic atherogenic LDL modifications enzymatically degraded LDL (E-LDL) and oxidized LDL (Ox-LDL) prepared from the same donor LDL. To detect differences in macrophage lipid storage, fluorescent high-content imaging was used. Lipid droplets were stained using Bodipy 493/503, and the fluorescent phospholipid probe NBD-PE was used to detect endolysosomal phospholipidosis in high-content imaging assays. The phospholipidosis assay was validated using phospholipidosis-inducing cationic amphiphilic drugs. In addition, neutral lipids and phospholipidosis were determined using LipidTOX. Images of 96-well cell culture microtiter plates were captured with multichannel laser-based high-content confocal microscopy, and subsequently cell- and well based data were analyzed. E-LDL-loaded macrophages show increased intensity of Bodipy 493/503 and LipidTOX-Green neutral lipid droplet staining and a greater mean area and number of lipid droplets per cell compared to Ox-LDL-loaded and M CSF-differentiated control macrophages. In contrast, Ox-LDL-loaded macrophages show increased intensity of NBD-PE and LipidTOX-Red detectable phospholipidosis in the endolysosomal compartment compared to E-LDL-loaded and M-CSF differentiated macrophages. Treatment with the peroxisome proliferator-activated receptor-gamma agonist pioglitazone leads to lipid droplet induction depending on the lipid loading state of the macrophages. These results indicate that E-LDL preferentially induces lipid droplets, while Ox-LDL provokes endolysosomal phospholipidosis in human macrophages representing two different lipid storage principles. Therefore, fluorescent high-content imaging is a useful tool to discriminate between and quantify lipid storage compartments in macrophages also in response to drugs affecting cellular lipid metabolism. PMID- 20014302 TI - Trends in genetic literature with the use of flow cytometry. PMID- 20014303 TI - Thyroid follicular neoplasm: analysis by fine needle aspiration cytology, frozen section, and histopathology. AB - We performed a retrospective analysis of follicular neoplasm data obtained from frozen section examinations of thyroid nodules. A total of 5,660 patients underwent preoperative neck ultrasonography and fine-needle aspiration cytology (FNAC), surgical treatment, and follow-up at a medical institute. Patients with papillary thyroid microcarcinoma were excluded from this study. In 971 cases, frozen section examination was performed during the surgical treatment of follicular neoplasm that was diagnosed via FNAC. Thyroid malignancies were histologically confirmed in 25.1% of cases (244/971). Among the patients with papillary thyroid carcinoma, 45 were diagnosed with the follicular variant of papillary thyroid carcinomas (27.4%). The diagnostic sensitivity of frozen section for the nonfollicular variant of papillary thyroid carcinoma was better than that for the follicular variant of papillary thyroid carcinoma (89.1% versus 78.9%; P = 0.1023). For 12 cases the diagnosis was atypical follicular adenomas. The diagnostic accuracy of frozen section in cases of follicular neoplasm was 76.9% with a sensitivity of 84.8% and a specificity of 98.9%. In conclusion, our analysis revealed high rates of accuracy when using frozen tissue sections for early diagnosis and treatment of follicular neoplasm; thus, an early decision to extent of surgery prevents a risky follow-up surgery. PMID- 20014305 TI - The prevalence of abnormal cervical cytology in women with infertility. AB - The aim of this study was to compare the frequency of abnormal cervical cytology in women with infertility problems with that of fertile women by using ThinPrep(r) liquid-based Pap TestsTM. A retrospective case-control study for over 2 years was conducted. The cases included all women with infertility problems who had Pap tests during their infertility treatment period. The cases were further subdivided into primary and secondary infertility groups. The control group included all women without infertility problems who had routine Pap tests in the same period. The age and demographic features were adjusted and matched for both groups. Statistical analysis included chi-square test and Fischer exact test. The infertility group (n = 490) showed significantly (P < 0.05) more abnormal cervical squamous epithelial abnormalities (48 positive cases, 9.8%) than the controls (n = 7,150, 216 positive cases, 3%). Women with secondary infertility had more epithelial abnormalities and more high-grade lesions than women with primary infertility. Women with infertility had statistically significant higher frequency of squamous intraepithelial lesions than women without infertility problems of similar age and demographic background. The data suggest that women with infertility might benefit from more frequent cervical cytology screening. PMID- 20014304 TI - Can mobile units improve the strategies for cervical cancer prevention? AB - Cervical cancer is a serious public health problem in women in developing countries because of absence or ineffectiveness of screening programs. Several biases to access medical care and inequity of public health system in a continental country like Brazil limit the implementation of adequate programs to appropriately prevent the cervical cancer. Therefore, the aim of this study was to evaluate the results of applying the mobile unit (MU) for cervical cancer screening. From May 2003 to May 2004, a cervical cancer screening was offered to women aged 20-69 years, residing in 19 municipal districts of the Barretos county region, in Sao Paulo. Out of the 9,560 examination available, 2,964 (31%) women underwent screening. The medium distance traveled by the MU was 45 km. The medium time spent by women in the MU for completion of the questionnaire and doing the exam was 20 minutes. It was observed that 17.0% of women screened had never had the test or had not had it repeated within the last 3 years. The negative response was more common among women aged 20 to 29 years and 60 to 69 years and among women with less schooling and lower socio-economic income (P < 0.05). MU can significantly overcome the chronic deficiency of public health system accessibility offering opportunity to these women to participate in screening programs. PMID- 20014306 TI - Metastatic signet ring cell carcinoma presenting as a thyroid nodule: report of a case with fine-needle aspiration cytology. AB - Metastatic carcinomas to the thyroid are quite rare in daily cytology practice. However, when present they may produce a diagnostic dilemma, particularly when they share some morphologic similarities with primary thyroid lesions and when occurring in patients with occult malignant history. Herein, we report a case of metastatic gastric signet ring cell carcinoma to the thyroid. Our patient presented with an isolated right thyroid nodule, which was clinically considered to be a primary thyroid neoplasm. Fine-needle aspiration (FNA) cytology of the nodule revealed a cellular specimen with cohesive fragments and scattered individual neoplastic cells. The neoplastic cells had enlarged nuclei, fine chromatin, and inconspicuous nucleoli. Nuclear crowding, molding, and grooving were prominent. Intranuclear inclusion-like clearance was identified. Some tumor cells also had eccentric nuclei, creating a signet ring cell appearance. The colloid was scant. These cytological features may be seen in cases of papillary thyroid carcinoma or signet ring cell follicular adenoma; however, the presence of the signet ring cells is unusual in primary thyroid lesions and raises the possibility of a metastatic lesion to the thyroid. In our case, the tumor cells were positive for AE1/AE3, mucicarmine, and periodic acid-Schiff, but negative for thyroglobulin and thyroid transcription factor-1. The patient was also found to have a 3.7-cm mass in the distal esophagus/proximal stomach. Biopsy of this mass showed an invasive signet ring cell carcinoma. The purpose of our study is to discuss the cytological features and the differential diagnosis of this unusual thyroid FNA case. PMID- 20014307 TI - Sarcomatoid collecting duct carcinoma of kidney diagnosed with urine and renal pelvic lavage cytology. AB - A case of sarcomatoid collecting duct carcinoma (CDC) of kidney is presented, in which the diagnosis was made cytologically with voided urine and renal pelvis lavage. Cytology of hemorrhagic voided urine revealed highly atypical adenocarcinoma cells with reminiscent ductal structure, which suggested CDC as the most likely diagnosis. Computed tomography and magnetic resonance imaging demonstrated a left renal tumor, and selective lavage of left renal pelvis yielded spindle-shaped, highly atypical cells that indicated sarcomatoid carcinoma. The diagnosis of renal cancer with urine cytology is challenging because of small number of tumor cells in the urine, which are often associated with degeneration. As the urinary cytologic findings of sarcomatoid CDC have not been reported, the characteristic cytologic findings of sarcomatoid CDC are described in detail, and the differential diagnoses with diagnostic pitfalls were discussed. PMID- 20014308 TI - Assessing for primary oropharyngeal or nasopharyngeal squamous cell carcinoma from fine needle aspiration of cervical lymph node metastases. AB - In fine needle aspirates of cervical lymph nodes with metastatic squamous cell carcinoma (SCC), the site of origin may not be clinically evident. The distinction between oropharyngeal and nasopharyngeal primary SCC has important management consequences. In the current study, we evaluated metastatic SCC for HPV types 16, 18, 31, 33, 51 (by in situ hybridization[ISH]), p16 and ProExC (surrogate HPV markers), and Epstein Barr Virus reported in nasopharyngeal SCC. Forty patients diagnosed between 2004 and 2008, with adequate cell block material were identified. ISH for high risk HPV and EBV (EBER), and immunohistochemistry for p16 and ProExC were performed. Primary site was designated in 31 cases with 26 head and neck including 11 oropharyngeal and 2 nasopharyngeal, and 5 other sites. High risk HPV was detected in 9 cases (22.5%), p16 in 16 (40%), ProExC in 35 (87.5%), and EBER in 2 (5%). All cases with high risk HPV ISH also showed overexpression of p16. The sensitivity for HPV infection by both surrogate markers was 100%; specificity for p16 and ProExC was 78.7 and 16.1%, respectively. Seven (63.6%) oropharyngeal SCC were positive for HPV ISH and negative for EBV; one nasopharyngeal SCC (50%) was EBER positive and HPV negative. HPV and EBER detection can serve as indicators for oropharyngeal and nasopharyngeal primary SCC, respectively, however our data show that only a subset (63.6%) of oropharyngeal SCC are high risk HPV-related. Additionally, despite their high sensitivity for HPV infection, surrogate markers, especially ProExC, lack specificity. PMID- 20014309 TI - Influence of defective bone marrow osteogenesis on fracture repair in an experimental model of senile osteoporosis. AB - Bone marrow osteogenesis in senile osteoporotic bone is impaired and, as such, may have significant implications on the successful outcome of fracture repair. Here we utilize a well-established murine model of senile osteoporosis, the P6 strain of senescence-accelerated mice (SAMP6), to investigate fracture healing in aged osteoporotic bone. A femoral osteotomy was created in SAMP6 and in non osteoporotic age-matched control R1 senescence-resistant mice (SAMR1). The course of fracture healing was evaluated over a period of 42 days using quantitative microCT and histological analysis. The differentiation capabilities of bone mesenchymal progenitor cells derived from SAMP6 and SAMR1 mice was examined, and their osteogenic potential determined. Although preliminary in vitro analysis confirmed that bone marrow-derived stem cells (BMSC) isolated from SAMP6 mice had a reduced osteogenic capacity, no significant deficit in fracture repair as determined by quantitative microCT could be detected. This was supported by histology assessment, where complete bridging of the fracture gap was evident by day 28 and was fully healed day 42 in both SAMP6 and SAMR1 mice. Further in vitro studies revealed that periosteal-derived progenitor cells (PDPC) isolated from SAMP6 mice had an osteogenic potential comparable to that observed in SAMR1 mice. In conclusion, fracture healing in SAMP6 mice is not detrimentally affected by impairment of BMSC osteogenesis, suggesting that bone marrow-mediated repair processes are dispensable for normal bone healing in this senile osteoporotic fracture model. Furthermore, the influence of PDPC in the repair process may partly explain the absence of any detectable deficits in fracture repair in SAMP6 mice. PMID- 20014310 TI - Kinetics of the UV-induced DNA damage response in relation to cell cycle phase. Correlation with DNA replication. AB - It has been reported that exposure to UV light triggers DNA damage response (DDR) seen as induction of gammaH2AX not only in S- but also in G(1)-phase cells. In the present study, in addition to gammaH2AX, we assessed other markers of DDR, namely phosphorylation of ATM on Ser1981, of ATM/ATR substrate on Ser/Thr at SQ/TQ cluster domains and of the tumor suppressor p53 on Ser15, in human pulmonary carcinoma A549 cells irradiated with 50 J/m(2) of UV-B. Phosphorylation of these proteins detected with phospho-specific Abs and measured by laser scanning cytometry in relation the cell cycle phase was found to be selective to S-phase cells. The kinetics of phosphorylation of ATM was strikingly similar to that of ATM/ATR substrate, peaking at 30 min after UV irradiation and followed by rapid dephosphorylation. The peak of H2AX phosphorylation was seen at 2 h and the peak of p53 phosphorylation at 4 h after exposure to UV light. Local high spatial density of these phospho-proteins reported by intensity of maximal pixel of immunofluorescence in the DDR nuclear foci was distinctly more pronounced in the early compared to late portion of S-phase. Exposure of cells to UV following 1 h pulse-labeling of their DNA with 5-ethynyl-2'deoxyuridine (EdU) made it possible to correlate the extent of DNA replication during the pulse with the extent of the UV-induced H2AX phosphorylation within the same cells. This correlation was very strong (R(2) = 0.98) and the cells that did not incorporate EdU showed no evidence of H2AX phosphorylation. The data are consistent with the mechanism in which stalling of DNA replication forks upon collision with the primary UV induced DNA lesions and likely formation of double-strand DNA breaks triggers DDR. The prior reports (including our own) on induction of gammaH2AX in G(1) cells by UV may have erroneously identified cells initiating DNA replication following UV exposure as G(1) cells due to the fact that their DNA content did not significantly differ from that of G(1) cells that had not initiated DNA replication. PMID- 20014311 TI - Atypical glandular cells (AGC): ThinPrep Imaging System (TIS), manual screening (MS), and correlation with Hybrid Capture 2 (HC2) HPV DNA testing. AB - OBJECTIVE: The aim of the study was to determine if the ThinPrep Imaging System (T1S) improves the positive predictive value (PPV) of atypical glandular cell (AGC) diagnosis for identifying HPV-related squamous and/or glandular lesions over manual screening (MS), and if human papilloma virus (HPV)-DNA testing improves the diagnostic yield. MATERIALS AND METHODS: 85 ThinPrep cervical cytology specimens with a diagnosis of AGC by TIS (n = 51) and MS (n = 34) were retrieved. The diagnoses were correlated with corresponding histologic follow-up and high risk (HR)-HPV testing results. RESULTS: The PPV of AGC by TIS and MS for HPV-related squamous lesions were similar. In the MS group, more cases of glandular pathology were identified, however only three represented adenocarcinoma in-situ (AIS), and the remaining ten were endometrial carcinomas (EMCA). CONCLUSIONS: TIS and MS are comparable in the detection of AGC representing squamous histology and the addition of HPV DNA testing does not differentially improve performance. Although the MS group harbored more glandular pathology, the differences in the detection of AIS were not statistically significant. PMID- 20014313 TI - Images in cytology. Sporangia, sporangium-like spherules and mimicking structures in respiratory cytopathology. PMID- 20014312 TI - Intraoperative cytology--role in bone lesions. AB - In spite of becoming an integral part of surgical pathology, very few reports are available regarding the utility of intraoperative cytology (IOC) exclusively for bone lesions. This study was undertaken in a view to fill this lacuna. Sixty bone lesions were evaluated intraoperatively with the help of cytology smears prepared by touch, scrape, or crush technique. The diagnosis made on cytological preparation was compared with histopathological diagnosis taking the latter as gold standard. Different parameters like reasons for Intraoperative consultation, best technique for preparation of smear, average time taken to render a diagnosis, and finally the accuracy of IOC was evaluated. Common reasons for the intraoperative consultation were to make or confirm a diagnosis for proper surgical intervention and to evaluate the surgical resection margin. Scrape was found to be the best method for cytological smear preparation. Average time taken to render a diagnosis was 20 minutes. Sensitivity, specificity, and overall diagnostic accuracy was 96.7, 96.6, and 96.6%, respectively. Cytology can play a valuable role in the intraoperative diagnosis of bone lesions. The method is simple, cheap, quick, and has no complication. It should be undertaken routinely, as a rapid intraoperative diagnosis will expedite timely and proper management of the patients, along with early post operative treatment and thus avoid the aggravating delays. PMID- 20014314 TI - Usefulness of NMP22 as an adjunct to a typical urine cytology and low-grade urothelial carcinoma. AB - The usefulness of urine cytology combined with NMP22 was evaluated for the primary diagnosis of urothelial carcinoma. Of 53 clinically suspected patients, histopathological diagnoses were low-grade urothelial carcinoma (25), high-grade urothelial carcinoma (13), and inflammatory lesions (15). Cytology was positive in 25 and negative in 14 patients. Fourteen of 25 low-grade urothelial carcinoma and 11/13 high-grade urothelial carcinoma were diagnosed correctly on urine cytology. Atypical cells seen in 14 patients were categorized as inconclusive for malignancy. The overall sensitivity of urine cytology was 65.8%, whereas specificity was 100%. NMP22 was positive in 33 patients. Of these 30, 18 low grade and 12 high-grade lesions were true positive. Of the 20 NMP22, eight negative cases were false-negative. Ten of 15 with negative histopathology were also negative for NMP22, three were false-positive, and two showed erratic results. Nine of 14 cases with atypical urine cytology were positive for NMP22. Eight of these showed low-grade carcinoma on histopathology. The sensitivity of BladderChek NMP22 test was 79%, whereas specificity was 80%. NMP22 BladderChek test is a useful adjunct to urine cytology in atypical and low-grade carcinoma. PMID- 20014315 TI - Correlation between ZAP-70, phospho-ZAP-70, and phospho-Syk expression in leukemic cells from patients with CLL. AB - For patients with chronic lymphocytic leukemia (CLL), expression of ZAP-70 in the leukemic cells is an indicator of poor prognosis. However, the mechanism that accounts for this effect is not known. ZAP-70 expression has previously been associated with increased B cell antigen receptor signaling upon surface immunoglobulin ligation in vitro as shown by ZAP-70 and Syk phosphorylation. This finding has led to the suggestion that a more aggressive clinical course is correlated with B cell antigen receptor signaling. Using high resolution immunophenotyping to analyze CLL cells ex vivo (without stimulation in vitro), we have demonstrated CLL cells from all patients express some ZAP-70 and that increased expression of ZAP-70 is correlated with decreased levels of phosphorylated ZAP-70 and phosphorylated Syk measured directly ex vivo. Conversely, high levels of phosphorylated ZAP-70 and phosphorylated Syk are found only in samples with low levels of ZAP-70 expression, and Syk and ZAP-70 phosphorylation appear to be mostly independent of each other. Additionally, Syk phosphorylation is directly correlated with levels of p21(cip1), a cell cycle inhibitor and a p53 target. Together these findings suggest that lower levels of p21(cip1) and/or a defect in p53 activity may account in part for the more aggressive disease course in patients with high levels of ZAP-70 rather than enhanced B cell antigen receptor signaling as has been previously hypothesized. PMID- 20014316 TI - Nerve compression activates selective nociceptive pathways and upregulates peripheral sodium channel expression in Schwann cells. AB - Chronic nerve compression (CNC) injuries, such as carpal tunnel syndrome, are common musculoskeletal conditions that affect patients with debilitating loss of sensory function and pain. Although early detection and treatment are important, our understanding of pain-related molecular mechanisms remains largely unclear. Here we investigate these mechanisms using an animal model for CNC injury. To confirm that CNC injury induces pain, we assessed expression of c-fos, a gene that is rapidly expressed in spinal sensory afferents in response to painful peripheral stimuli, and TNF-alpha and IL-6, two proinflammatory cytokines that are crucial to development of inflammatory-mediated pain. Results show c-fos upregulation 1-2 weeks postinjury in the absence of TNF-alpha or IL-6 expression, indicating increased neural sensitivity without an inflammatory response. This is consistent with previous studies that showed no morphologic evidence of inflammation in the CNC model. Surprisingly, we also found de novo expression of Na(V)1.8, a sodium channel linked to the development of neuropathic pain, in endoneurial Schwann cells following injury. Until now, Na(V)1.8 expression was thought to be restricted to sensory neurons. CNC injury appears to be a unique model of noninflammatory neuropathic pain. Further investigation of the underlying molecular basis could yield promising targets for early diagnosis and treatment. PMID- 20014319 TI - Strontium ranelate treatment enhances hydroxyapatite-coated titanium screws fixation in osteoporotic rats. AB - Increased bone turnover with excessive bone resorption and decreased bone formation is known to impair implant fixation. Strontium ranelate is well known as an effective antiosteoporotic agent by its dual effect of antiresorbing and bone-forming activity. This study was designed to evaluate the effect of systemic strontium ranelate (SR) treatment on fixation of hydroxyapatite (HA)-coated titanium screws in ovariectomized (OVX) rats. Twelve weeks after being OVX (n=30) or sham (n=10) operated, 40 female Sprague-Dawley rats received unilateral implants in the proximal tibiae. The OVX rats were randomly divided into the following groups: OVX, OVX+SRL ("L" refers to low SR dose of 500 mg/kg/day), OVX+SRH ("H" refers to high SR dose of 1000 mg/kg/day).Twelve weeks after treatment, bone blocks with implants were evaluated with micro-CT and biomechanical push-out tests. Compared to OVX animals, SR treatment increased the bone volume ratio by 51.5% and 1.1-fold, the percentage osteointegration by 1.0 fold and 1.9-fold in micro-CT evaluation, and the maximal force by 1.9-fold and 3.3-fold in biomechanical push-out test, for the low and high dose of SR, respectively. Significant correlation between micro-CT and biomechanical properties demonstrated that trabecular parameters played an important role in predicting the biomechanical properties of implant fixation. Our findings suggest that SR treatment can dose-dependently improve HA-coated screw fixation in OVX rats and facilitate the stability of the implant in the osteoporotic bone. PMID- 20014320 TI - Polymethylmethacrylate particles impair osteoprogenitor viability and expression of osteogenic transcription factors Runx2, osterix, and Dlx5. AB - Polymethylmethacrylate (PMMA) particles have been shown to inhibit the differentiation of osteoprogenitor cells, but the mechanism of this inhibitory effect has not been investigated. We hypothesize that the inhibitory effects of PMMA particles involve impairment of osteoprogenitor viability and direct inhibition of transcription factors that regulate osteogenesis. We challenged MC3T3-E1 osteoprogenitors with PMMA particles and examined the effects of these materials on osteoprogenitor viability and expression of transcription factors Runx2, osterix, Dlx5, and Msx2. MC3T3-E1 cells treated with PMMA particles over a 72-h period showed a significant reduction in cell viability and proliferation as indicated by a dose- and time-dependent increase in supernatant levels of lactate dehydrogenase, an intracellular enzyme released from dead cells, a dose-dependent decrease in cell number and BrdU uptake, and the presence of large numbers of positively labeled Annexin V-stained cells. The absence of apoptotic cells on TUNEL assay indicated that cell death occurred by necrosis, not apoptosis. MC3T3 E1 cells challenged with PMMA particles during the first 6 days of differentiation in osteogenic medium showed a significant dose-dependent decrease in the RNA expression of Runx2, osterix, and Dlx5 on all days of measurement, while the RNA expression of Msx2, an antagonist of Dlx5-induced osteogenesis, remained relatively unaffected. These results indicate that PMMA particles impair osteoprogenitor viability and inhibit the expression of transcription factors that promote osteoprogenitor differentiation. PMID- 20014321 TI - Evolution of a precursor. PMID- 20014322 TI - The current status of audiologic rehabilitation for profound unilateral sensorineural hearing loss. AB - OBJECTIVES/HYPOTHESIS: Audiologic rehabilitation of individuals with profound unilateral sensorineural hearing loss (USNHL) has traditionally been limited to the use of air-conduction contralateral routing of sound (CROS) hearing aids. Treatment for these individuals has expanded with new applications of the bone anchored hearing aid (BAHA), transcranial hearing aid (t-CROS), and the cochlear implant. In this article, the authors review the literature that addresses these various treatment options. STUDY DESIGN: Contemporary review RESULTS: Historical information is available that describes the limited efficacy of air-conduction CROS hearing aids in lifting hearing handicap associated with USNHL. Current investigations on providing cross hearing are generally focused on use of the BAHA. Little is known at present whether new developments in hearing aid technology can improve on conventional air-conduction CROS or t-CROS approaches. Interestingly, the cochlear implant seems to be a viable option for individuals with USNHL and tinnitus who also have intact auditory nerve pathways. CONCLUSIONS: There is indication in the literature that BAHA provides greater relief of hearing handicap associated with USNHL than CROS hearing aids; however, both have been found to provide limited patient satisfaction and seemingly fall short of restoring true sound localization. Adequate trials have not been performed comparing BAHA with the best CROS hearing aid technology. Transcranial hearing aids and cochlear implants are experimental methods to treat USNHL and hold promise, although there remains a lack of studies available to fully support this. PMID- 20014323 TI - Model of cap-dependent translation initiation in sea urchin: a step towards the eukaryotic translation regulation network. AB - The large and rapid increase in the rate of protein synthesis following fertilization of the sea urchin egg has long been a paradigm of translational control, an important component of the regulation of gene expression in cells. This translational up-regulation is linked to physiological changes that occur upon fertilization and is necessary for entry into first cell division cycle. Accumulated knowledge on cap-dependent initiation of translation makes it suited and timely to start integrating the data into a system view of biological functions. Using a programming environment for system biology coupled with model validation (named Biocham), we have built an integrative model for cap-dependent initiation of translation. The model is described by abstract rules. It contains 51 reactions involved in 74 molecular complexes. The model proved to be coherent with existing knowledge by using queries based on computational tree logic (CTL) as well as Boolean simulations. The model could simulate the change in translation occurring at fertilization in the sea urchin model. It could also be coupled with an existing model designed for cell-cycle control. Therefore, the cap-dependent translation initiation model can be considered a first step towards the eukaryotic translation regulation network. PMID- 20014324 TI - Slightly modifying pseudoproline dipeptides incorporation strategy enables solid phase synthesis of a 54 AA fragment of caveolin-1 encompassing the intramembrane domain. AB - This work contributes to highlight the benefits of pseudoproline dipeptides introduction in difficult SPPS. We show how a slight modification in the positioning choice conditioned the synthesis achievement of a 54 amino acid long caveolin-1 peptide encompassing the intramembrane domain. Furthermore, we report a side reaction correlated with the coupling steps and generating truncated fragments with a mass deviation of + 42 Da. Considering the need of structural data for membrane proteins, most of which are considered as prevalent therapeutic targets, chemical synthesis provides an interesting alternative pathway to obtain hydrophobic domains by pushing back the frontiers of conventional RP methods of purification. PMID- 20014325 TI - Design, synthesis and biological activity of cell-penetrating peptide-modified octreotide analogs. AB - Four novel octreotide analogs with cell-penetrating peptides (CPPs) at the N terminus or C-terminus were synthesized by a stepwise Fmoc solid-phase synthesis strategy. The synthesized peptides were analyzed and characterized using reverse phase HPLC and MALDI-TOF mass spectrometry. The antiproliferative activity of the analogs was tested in vitro on human gastric (SGC-7901) and hepatocellular cancer (BEL7402) cell lines using the 3-(4,5-dimethylthiazol-2-yl)-2,5 diphenyltetrazolium bromide (MTT) assay. Interestingly, these analogs showed a higher anticancer activities than the parent octreotide except CMTPT03 analog. The results demonstrate that the designed octreotide analogs enhance their anticancer activity after linking together the CPPs to octreotide at the N terminus, and are potential molecules for future use in cancer therapy and drug targeting. PMID- 20014326 TI - Transvaginal cervical length measurement for prediction of preterm birth in women with threatened preterm labor: a meta-analysis. AB - OBJECTIVES: To integrate data on the performance of cervical length measurement for the prediction of preterm birth in symptomatic women. METHODS: MEDLINE, SCOPUS and manual searches for studies with transvaginal ultrasound measurement of the cervical length in symptomatic women were carried out. Random effects models were used for data integration, and pooled test estimates of sensitivity, specificity, and positive and negative likelihood ratios (LR+ and LR-) were calculated along with their 95% CIs. RESULTS: Twenty-eight studies fulfilled the selection criteria. For birth within 1 week from presentation, the pooled sensitivity, specificity, LR+ and LR- of cervical length < 15 mm were 59.9% (95% CI, 52.7-66.8%), 90.5% (95% CI, 89.0-91.9%), 5.71 (95% CI, 3.77-8.65) and 0.51 (95% CI, 0.33-0.80), respectively. The same estimates for studies with presentation at or before 34 + 0 weeks were 71.0% (95% CI, 60.6-79.9%), 89.8% (95% CI, 87.4-91.9%), 5.19 (95% CI, 2.29-11.74) and 0.38 (95% CI, 0.11-1.34), respectively. For prediction of birth before 34 weeks, the pooled sensitivity, specificity, LR+ and LR- of cervical length < 15 mm were 46.2% (95% CI, 34.8 57.8%), 93.7% (95% CI, 90.7-96.0%), 4.31 (95% CI, 2.73-6.82) and 0.63 (95% CI, 0.38-1.04), respectively. There was considerable heterogeneity across studies in most estimates. CONCLUSIONS: Measurement of cervical length in symptomatic women can detect a significant proportion of those who will deliver within 1 week and help to rationalize their management. The considerable heterogeneity across studies may be indicative of methodological flaws, which either were not reported at all or were under-reported. PMID- 20014327 TI - Diagnosis and management of vasa previa: a questionnaire survey. AB - OBJECTIVES: Our aim was to assess the current use of obstetric ultrasound imaging for the diagnosis of asymptomatic vasa previa. We also investigated obstetricians' views on the feasibility of a screening policy and their awareness of risk factors associated with this condition. METHODS: A national postal survey was conducted between March and July 2006. A total of 234 questionnaires were sent to obstetric and fetomaternal consultants across England and Wales. In all, 128 questionnaires were returned, a response rate of 55%. RESULTS: Most respondents (85%) stated that in their hospital they do not report velamentous cord insertions at the anomaly scan. However, 73% occasionally or routinely document the presence of succenturiate lobes. Only 33% of respondents offered transvaginal scanning for the identification of vasa previa within their hospital, whereas only 6% had ever referred women to a tertiary center for this indication. In all, 34% of the respondents did not identify any risk factor for the condition. Most respondents (80%) would offer an elective Cesarean section if vasa previa was suspected antenatally; the majority would perform it at 38 weeks' gestation. However, only 20% of respondents felt that an effective screening policy is possible. CONCLUSIONS: Despite evidence that perinatal death can be prevented by antenatal diagnosis of vasa previa, most obstetricians in England and Wales feel that a screening policy is not possible. The majority of them would offer an elective Cesarean section for vasa previa at around 38 weeks. There is a need to increase awareness and understanding of the major risk factors for this condition. PMID- 20014328 TI - Three-dimensional multiplanar ultrasound is a valuable tool in the study of the fetal profile in the second trimester of pregnancy. AB - OBJECTIVES: To evaluate the additional value of three-dimensional (3D) multiplanar ultrasound in the examination of the fetal profile. METHODS: Two 3D volumes of the fetal head were obtained from 84 fetuses at 22 to 29 weeks' gestation. The volumes were taken starting at the midsagittal plane with the fetus facing the transducer. The success rate and acquisition time to obtain each volume and display the exact midsagittal plane by 3D multiplanar ultrasound were analyzed. The correction angles from the original two-dimensional (2D) profile view to the exact midsagittal plane were noted. Of six measurements, related to the fetal nose and jaws, the success rate and the intraobserver reproducibility between the 2D and the 3D multiplanar ultrasound were compared. RESULTS: In 81 (96.4%) cases we succeeded in obtaining a profile volume, 70% of the volumes being obtained within 10 min. It was possible to define by multiplanar mode the exact midsagittal plane in less than 1 min. The mean rotation necessary to obtain the exact midsagittal plane with 3D multiplanar mode was significantly larger around the y-axis (11.9 degrees ) than around the z-axis (4.3 degrees ) of the fetus. For between 5 and 12% of the six measurements under investigation it was not possible to obtain values with 2D ultrasound. However, 3D ultrasound made these measurements possible in at least one volume. The intraobserver reproducibility was higher with 3D multiplanar ultrasound than with 2D ultrasound, this difference being statistically significant for five of the six measurements. CONCLUSIONS: 3D multiplanar ultrasound improves the topographic depiction of the midsagittal profile view, enables correct measurement of anatomical details and improves intraobserver reproducibility. 3D multiplanar ultrasound is a powerful instrument for investigating the fetal profile. PMID- 20014329 TI - Fetal echocardiography: z-score reference ranges for a large patient population. AB - OBJECTIVES: The main goal was to develop new z-score reference ranges for common fetal echocardiographic measurements from a large referral population. METHODS: A retrospective cross-sectional study of 2735 fetuses was performed for standard biometry (biparietal diameter (BPD) and femoral diaphysis length (FDL)) and an assessment of menstrual age (MA). Standardized fetal echocardiographic measurements included aortic valve annulus and pulmonary valve annulus diameters at end-systole, right and left ventricular diameters at end-diastole, and cardiac circumference from a four-chamber view of the heart during end-diastole. Normal z score ranges were developed for these echocardiographic measurements using MA, BPD and FDL as independent variables. This was accomplished by using first standard regression analysis and then weighted regression of absolute residual values for each parameter in order to adjust for inconstant variance. RESULTS: A simple, linear regression model was the best description of the data in each case and correlations between fetal cardiac measurements and the independent variables were excellent. There was significant heteroscedasticity of standard deviation with increasing gestational age, which also could be modeled with simple linear regression. After this adjustment, the residuals conformed to a normal distribution, validating the calculation and interpretation of z-scores. CONCLUSION: Development of reliable z-scores is possible for common fetal echocardiographic parameters by applying statistical methods that are based on a large sample size and weighted regression of absolute residuals in order to minimize the effect of heteroscedasticity. These normative ranges should be especially useful for the detection and monitoring of suspected fetal cardiac size and growth abnormalities. PMID- 20014330 TI - Retronasal triangle: a sonographic landmark for the screening of cleft palate in the first trimester. AB - OBJECTIVES: To describe a new first-trimester sonographic landmark, the retronasal triangle, which may be useful in the early screening for cleft palate. METHODS: The retronasal triangle, i.e. the three echogenic lines formed by the two frontal processes of the maxilla and the palate visualized in the coronal view of the fetal face posterior to the nose, was evaluated prospectively in 100 consecutive normal fetuses at the time of routine first-trimester sonographic screening at 11 + 0 to 13 + 6 weeks' gestation. In a separate study of five fetuses confirmed postnatally as having a cleft palate, ultrasound images, including multiplanar three-dimensional views, were analyzed retrospectively to review the retronasal triangle. RESULTS: None of the fetuses evaluated prospectively was affected by cleft lip and palate. During their first-trimester scan, the retronasal triangle could not be identified in only two fetuses. Reasons for suboptimal visualization of this area included early gestational age at scanning (11 weeks) and persistent posterior position of the fetal face. Of the five cases with postnatal diagnosis of cleft palate, an abnormal configuration of the retronasal triangle was documented in all cases on analysis of digitally stored three-dimensional volumes. CONCLUSIONS: This study demonstrates the feasibility of incorporating evaluation of the retronasal triangle into the routine evaluation of the fetal anatomy at 11 + 0 to 13 + 6 weeks' gestation. Because fetuses with cleft palate have an abnormal configuration of the retronasal triangle, focused examination of the midface, looking for this area at the time of the nuchal translucency scan, may facilitate the early detection of cleft palate in the first trimester. PMID- 20014331 TI - Study of a lipophilic captopril analogue binding to angiotensin I converting enzyme. AB - Human ACE is a central component of the renin-angiotensin system and a major therapeutic target for cardiovascular diseases. The somatic form of the enzyme (sACE) comprises two homologous metallopeptidase domains (N and C), each bearing a zinc active site with similar but distinct substrate and inhibitor specificities. In this study, we present the biological activity of silacaptopril, a silylated analogue of captopril, and its binding affinity towards ACE. Based on the recently determined crystal structures of both the ACE domains, a series of docking calculations were carried out in order to study the structural characteristics and the binding properties of silacaptopril and its analogues with ACE. PMID- 20014332 TI - Metabolic syndrome and risk of development of chronic kidney disease: the Niigata preventive medicine study. AB - BACKGROUND: The metabolic syndrome consists of a cluster of cardiovascular risk factors, many of which have also been implicated in the genesis of chronic kidney disease. We studied the impact of the metabolic syndrome on chronic kidney disease in general population. METHODS: The Niigata Preventive Medicine Study was community-based prospective observational cohort study based upon the annual health examinations in Japan. We studied the association of the metabolic syndrome with a risk of development of kidney dysfunction and proteinuria in 34 986 participants without baseline kidney disease. RESULTS: The metabolic syndrome was present in 3679 subjects (11%). During a follow-up of 5.8 years, kidney dysfunction developed in 184 subjects with metabolic syndrome (5.0%) and 746 subjects without metabolic syndrome (2.4%). The metabolic syndrome was associated with development of kidney dysfunction (hazard ratio [HR], 2.12). All of the metabolic syndrome components were associated with risk of kidney dysfunction. The risk of kidney dysfunction increased across a number of the fulfilled metabolic syndrome components. The association of metabolic syndrome with kidney dysfunction remained significant in subjects without hypertension, diabetes, or cardiovascular disease (HR, 1.99) and in those < or =60 years without hypertension, diabetes, or cardiovascular disease (HR, 2.11). The metabolic syndrome was similarly associated with the development of proteinuria in all subjects (HR, 1.67), in those without hypertension, diabetes, or cardiovascular disease (HR, 1.64) and in those < or =60 years without hypertension, diabetes, or cardiovascular disease (HR, 2.14). CONCLUSIONS: The metabolic syndrome was associated with kidney disease even in subjects without major classical risk factors for chronic kidney disease. PMID- 20014334 TI - Genomic localization of the Z/EG transgene in the mouse genome. AB - The Z/EG transgenic mouse line, produced by Novak et al., displays tissue specific EGFP expression after Cre-mediated recombination. The autofluorescence of EGFP allows the visualization of cells of interest displaying Cre recombination. The initial construct was designed such that cells without Cre recombination express the beta-galactosidase marker, facilitating counterselection. We used inverse PCR to identify the site of integration of the Z/EG transgene, to improve the efficiency of homozygous Z/EG mouse production. Recombined cells produced large amounts of EGFP protein, resulting in higher levels of fluorescence and therefore greater contrast with nonrecombined cells. We mapped the transgene to the G1 region of chromosome 5. This random insertion was found to have occurred 230-bp upstream from the start codon of the Rasa4 gene. The insertion of the Z/EG transgene in the C57BL/6 genetic background had no effect on Rasa4 expression. Homozygous Z/EG mice therefore had no obvious phenotype. PMID- 20014333 TI - Image-guided radio-frequency gain calibration for high-field MRI. AB - High-field (>= 3T) MRI provides a means to increase the signal-to-noise ratio, due to its higher tissue magnetization compared with 1.5T. However, both the static magnetic field (B(0)) and the transmit radio-frequency (RF) field (B 1+) inhomogeneities are comparatively higher at higher field strengths than those at 1.5T. These challenging factors at high-field strengths make it more difficult to accurately calibrate the transmit RF gain using standard RF calibration procedures. An image-based RF calibration procedure was therefore developed, in order to accurately calibrate the transmit RF gain within a specific region-of interest (ROI). Using a turbo fast low-angle shot (TurboFLASH) pulse sequence with centric k-space reordering, a series of 'saturation-no-recovery' images was acquired by varying the flip angle of the preconditioning pulse. In the resulting images, the signal null occurs in regions where the flip angle of the preconditioning pulse is 90 degrees . For a given ROI, the mean signal can be plotted as a function of the nominal flip angle, and the resulting curve can be used to quantitatively identify the signal null. This image-guided RF calibration procedure was evaluated through phantom and volunteer imaging experiments at 3T and 7T. The image-guided RF calibration results in vitro were consistent with standard B(0) and B 1+ maps. The standard automated RF calibration procedure produced approximately 20% and 15-30% relative error in the transmit RF gain in the left kidney at 3T and brain at 7T, respectively. For initial application, a T(2) mapping pulse sequence was applied at 7T. The T(2) measurements in the thalamus at 7T were 60.6 ms and 48.2 ms using the standard and image-guided RF calibration procedures, respectively. This rapid, image-guided RF calibration procedure can be used to optimally calibrate the flip angle for a given ROI and thus minimize measurement errors for quantitative MRI and MR spectroscopy. PMID- 20014335 TI - The genesis of new and exciting developmental genetics research. PMID- 20014336 TI - Detection of cancer in cervical tissue biopsies using mobile lipid resonances measured with diffusion-weighted (1)H magnetic resonance spectroscopy. AB - The purpose of this study was to implement a diffusion-weighted sequence for visualisation of mobile lipid resonances (MLR) using high resolution magic angle spinning (HR-MAS) (1)H MRS and to evaluate its use in establishing differences between tissues from patients with cervical carcinoma that contain cancer from those that do not. A stimulated echo sequence with bipolar gradients was modified to allow T(1) and T(2) measurements and optimised by recording signal loss in HR MAS spectra as a function of gradient strength in model lipids and tissues. Diffusion coefficients, T(1) and apparent T(2) relaxation times were measured in model lipid systems. MLR profiles were characterised in relation to T(1) and apparent T(2) relaxation in human cervical cancer tissue samples. Diffusion weighted (DW) spectra of cervical biopsies were quantified and peak areas analysed using linear discriminant analysis (LDA). The optimised sequence reduced spectral overlap by suppressing signals originating from low molecular weight metabolites and non-lipid contributions. Significantly improved MLR visualisation allowed visualisation of peaks at 0.9, 1.3, 1.6, 2.0, 2.3, 2.8, 4.3 and 5.3 ppm. MLR analysis of DW spectra showed at least six peaks arising from saturated and unsaturated lipids and those arising from triglycerides. Significant differences in samples containing histologically confirmed cancer were seen for peaks at 0.9 (p < 0.006), 1.3 (p < 0.04), 2.0 (p < 0.03), 2.8 (p < 0.003) and 4.3 ppm (p < 0.0002). LDA analysis of MLR peaks from DW spectra almost completely separated two clusters of cervical biopsies (cancer, 'no-cancer'), reflecting underlying differences in MLR composition. Generated Receiver Operating Characteristic (ROC) curves and calculated area under the curve (0.962) validated high sensitivity and specificity of the technique. Diffusion-weighting of HR-MAS spectroscopic sequences is a useful method for characterising MLR in cancer tissues and displays an accumulation of lipids arising during tumourigenesis and an increase in the unsaturated lipid and triglyceride peaks with respect to saturated MLR. PMID- 20014337 TI - TDP-43, a neuro-pathosignature factor, is essential for early mouse embryogenesis. AB - TDP-43 is a highly conserved and ubiquitously expressed nuclear protein. It has been implicated in the regulation of transcription, alternative splicing, translation, and neuronal plasticity. TDP-43 has also been shown to be a disease signature protein associated with several neurodegenerative diseases including amyotrophic lateral sclerosis. However, the correlation of the physiological functions of TDP-43 with these diseases remains unknown. We have used the gene targeting approach to disrupt the expression of TDP-43 in mouse. Loss of the TDP 43 expression results in peri-implantation lethality of mice between embryonic days (E) 3.5 and 6.5. Blastocysts of the homozygous Tardbp null mutants are morphologically normal, but exhibit defective outgrowth of the inner cell mass in vitro. Our data demonstrate the essential function of TDP-43 in peri-implantation stage during the embryo development, likely because of its involvement in multiple biological processes in a variety of cell types. PMID- 20014342 TI - The zebrafish dyrk1b gene is important for endoderm formation. AB - Nodal-signaling is required for specification of mesoderm, endoderm, establishing left-right asymmetry, and craniofacial development. Wdr68 is a WD40-repeat domain containing protein recently shown to be required for endothelin-1 (edn1) expression and subsequent lower jaw development. Previous reports detected the Wdr68 protein in multiprotein complexes containing mammalian members of the dual specificity tyrosine-regulated kinase (dyrk) family. Here we describe the characterization of the zebrafish dyrk1b homolog. We report the detection of a physical interaction between Dyrk1b and Wdr68. We also found perturbations of nodal signaling in dyrk1b antisense morpholino knockdown (dyrk1b-MO) animals. Specifically, we found reduced expression of lft1 and lft2 (lft1/2) during gastrulation and a near complete loss of the later asymmetric lft1/2 expression domains. Although wdr68-MO animals did not display lft1/2 expression defects during gastrulation, they displayed a near complete loss of the later asymmetric lft1/2 expression domains. While expression of ndr1 was not substantially effected during gastrulation, ndr2 expression was moderately reduced in dyrk1b-MO animals. Analysis of additional downstream components of the nodal signaling pathway in dyrk1b-MO animals revealed modestly expanded expression of the dorsal axial mesoderm marker gsc while the pan-mesodermal marker bik was largely unaffected. The endodermal markers cas and sox17 were also moderately reduced in dyrk1b-MO animals. Notably, and similar to defects previously reported for wdr68 mutant animals, we also found reduced expression of the pharyngeal pouch marker edn1 in dyrk1b-MO animals. Taken together, these data reveal a role for dyrk1b in endoderm formation and craniofacial patterning in the zebrafish. PMID- 20014343 TI - Probing the photothermal effect of gold-based nanocages with surface-enhanced Raman scattering (SERS). PMID- 20014344 TI - Evidence that the C-terminal domain (CtD) autoinhibits neural repression by Drosophila E(spl)M8. AB - Analysis of the retinal defects of a CK2 phosphomimetic variant of E(spl)M8 (M8S(159)D) and the truncated protein M8* encoded by the E(spl)D allele, suggest that the nonphosphorylated CtD "autoinhibits" repression. We have investigated this model by testing for inhibition (in "trans") by the CtD fragment in its nonphosphorylated (M8-CtD) and phosphomimetic (M8SD-CtD) states. In N(+) flies, ectopic M8-CtD compromises lateral inhibition, i.e., elicits supernumerary bristles as with loss of N signaling. This antimorphic activity of M8-CtD strongly rescues the reduced eye and/or bristle loss phenotypes that are elicited by ectopic M8SD or wild type M8. Additionally, the severely reduced eye of N(spl)/Y; E(spl)D/+ flies is also rescued by M8-CtD. Rescue is specific to the time and place, the morphogenetic furrow, where "founding" R8 photoreceptors are specified. In contrast, the phosphomimetic M8SD-CtD that is predicted to be deficient for autoinhibition, exhibits significantly attenuated or negligible activity. These studies provide evidence that autoinhibition by the CtD regulates M8 activity in a phosphorylation-dependent manner. PMID- 20014345 TI - Inducible cardiomyocyte-specific gene disruption directed by the rat Tnnt2 promoter in the mouse. AB - We developed a conditional and inducible gene knockout methodology that allows effective gene deletion in mouse cardiomyocytes. This transgenic mouse line was generated by coinjection of two transgenes, a "reverse" tetracycline-controlled transactivator (rtTA) directed by a rat cardiac troponin T (Tnnt2) promoter and a Cre recombinase driven by a tetracycline-responsive promoter (TetO). Here, Tnnt2 rtTA activated TetO-Cre expression takes place in cardiomyocytes following doxycycline treatment. Using two different mouse Cre reporter lines, we demonstrated that expression of Cre recombinase was specifically and robustly induced in the cardiomyocytes of embryonic or adult hearts following doxycycline induction, thus, allowing cardiomyocyte-specific gene disruption and lineage tracing. We also showed that rtTA expression and doxycycline treatment did not compromise cardiac function. These features make the Tnnt2-rtTA;TetO-Cre transgenic line a valuable genetic tool for analysis of spatiotemporal gene function and cardiomyocyte lineage tracing during developmental and postnatal periods. PMID- 20014349 TI - Identification and functionality of proteomes secreted by rat cardiac stem cells and neonatal cardiomyocytes. AB - In the heart, the proteomes secreted by both cardiac stem cells (CSCs) and cardiac myocytes could act synergistically, but the identification and functionality of the proteins comprising the individual secretomes have not yet been described. In this study, we have identified proteins present in the media obtained from cultured rat CSCs and from cultured neonatal rat ventricular myocytes (NRVMs) and compared them with proteins identified in the media alone. Briefly, 83 unique proteins were identified after analysis by RPLC and MS. In total 49 and 23% were NRVM-specific or CSC-specific proteins, respectively, and 63% of total 83 proteins were integral plasma membrane and/or known secreted proteins. Fifteen proteins met our criteria for paracrine/autocrine factors: (i) robust protein identification, (ii) cell specific and (iii) known to be secreted. Most of these proteins have not been previously linked to stem cells. NRVM specific proteins atrial natriuretic factor (ANP) and connective tissue growth factor, and CSC-specific protein interleukin-1 receptor-like 1 (ST2) were found to affect rat CSC proliferation. These findings suggest that relative concentration of each protein may be crucial for cellular intertalk and for the final outcome of cardiac cell therapy. PMID- 20014350 TI - Hypertonic saline or high volume normal saline for viral bronchiolitis: mechanisms and rationale. PMID- 20014351 TI - Pediatric respiratory medicine--an international perspective. AB - Although Pediatric Respiratory Medicine as a subspecialty has a long tradition and is well established in some countries, there is a wide variation across different regions of the world with regard to e.g. recognition of the discipline, training requirements, training facilities and clinical needs. This review summarizes the situation in North America (US and Canada), South America, Asia, Australia, Israel and Europe with the aim to highlight commonalities and differences and, ultimately, to further support continuous development of paediatric Respiratory Medicine Worldwide. PMID- 20014352 TI - Tackling age-related disease: time for a step forward. PMID- 20014353 TI - Sample size requirement to detect an intervention effect at the end of follow-up in a longitudinal cluster randomized trial. AB - It is often anticipated in a longitudinal cluster randomized clinical trial (cluster-RCT) that the course of outcome over time will diverge between intervention arms. In these situations, testing the significance of a local intervention effect at the end of the trial may be more clinically relevant than evaluating overall mean differences between treatment groups. In this paper, we present a closed-form power function for detecting this local intervention effect based on maximum likelihood estimates from a mixed-effects linear regression model for three-level continuous data. Sample size requirements for the number of units at each data level are derived from the power function. The power function and the corresponding sample size requirements are verified by a simulation study. Importantly, it is shown that sample size requirements computed with the proposed power function are smaller than that required when testing group mean difference using data only at the end of trial and ignoring the course of outcome over the entire study period. PMID- 20014355 TI - Estimation of the effect size in meta-analysis with few studies. AB - Meta-analysis is often conducted with only a small number of studies. Adjustments of the (restricted) maximum likelihood estimator of the effect size are derived and their gains in efficiency are explored. The proposed estimators are applied to three sets of studies. PMID- 20014354 TI - Patient death as a censoring event or competing risk event in models of nursing home placement. AB - Participant death is often observed in studies that examine predictors of events, such as hospitalization or institutionalization, in older adult populations. The Cox proportional hazards modeling of the target event, whereby death is treated as a censoring event, is the standard analysis in this competing risks situation. However, the assumption of noninformative censoring applied to a frequently occurring competing event like death may be invalid and complicate interpretation in terms of the probability of the event. Multiple cause-specific hazard (CSH) models can be estimated, but ambiguities may arise when interpreting covariate effects across multiple CSH models and in terms of the cumulative incidence function (CIF). Alternatively, one can model the proportional hazards of the subdistribution of the CIF and evaluate the covariate effects on the CIF directly. We examine and compare these two approaches with nursing home (NH) placement data from a randomized controlled trial of a counseling and support intervention for spouse-caregivers of patients with Alzheimer's disease. CSHs for NH placement (where death is treated as a censoring event) and death (where NH placement is treated as a censoring event) and subdistribution hazards of the CIF for NH placement are modeled separately. In the presence of multiple covariates, the intervention effect is significant in both approaches, but the interpretation of the covariate effects requires joint evaluation of all estimated models. PMID- 20014356 TI - Application of the Bayesian dynamic survival model in medicine. AB - The Bayesian dynamic survival model (BDSM), a time-varying coefficient survival model from the Bayesian prospective, was proposed in early 1990s but has not been widely used or discussed. In this paper, we describe the model structure of the BDSM and introduce two estimation approaches for BDSMs: the Markov Chain Monte Carlo (MCMC) approach and the linear Bayesian (LB) method. The MCMC approach estimates model parameters through sampling and is computationally intensive. With the newly developed geoadditive survival models and software BayesX, the BDSM is available for general applications. The LB approach is easier in terms of computations but it requires the prespecification of some unknown smoothing parameters. In a simulation study, we use the LB approach to show the effects of smoothing parameters on the performance of the BDSM and propose an ad hoc method for identifying appropriate values for those parameters. We also demonstrate the performance of the MCMC approach compared with the LB approach and a penalized partial likelihood method available in software R packages. A gastric cancer trial is utilized to illustrate the application of the BDSM. PMID- 20014357 TI - Is pediatric pulmonary research training in dire straits. PMID- 20014358 TI - Extent of pharmacovigilance among resident doctors in Edo and Lagos states of Nigeria. AB - BACKGROUND: Systematic attention to adverse drug reactions (ADRs) started with the thalidomide disaster in the early 60s of the 20(th) century. In 1968 ten countries supported a spontaneous reporting system for ADR and collaborated with the WHO pilot research project for international drug monitoring. This survey was conducted to determine the level of awareness of resident doctors about ADRs reporting, the extent of their involvement in pharmacovigilance activities and to build a useful data base that will improve the quality of health care in Nigeria. SETTING: This cross-sectional observational survey was conducted in University of Benin Teaching Hospital (UBTH) and Irrua Specialist Teaching Hospital (ISTH), Lagos University Teaching Hospital (LUTH) and Lagos State University Teaching Hospital (LASUTH) in Edo and Lagos states, Nigeria, respectively. STUDY DESIGN / METHODS: A cross-sectional observational survey was used in this study. Total population under survey being 971 residents with the sample size of 350 statistically determined. RESULTS: The survey conducted revealed that 78.1% (258) of the respondents had inadequate knowledge about pharmacovigilance with 71.2% (235) unaware of the yellow forms for ADR reporting distributed by the National Agency for Food and Drug Administration and Control (NAFDAC). The respondents (92.4%) have observed ADRs in the course of their training and practice, yet only 25.5% of cases were reported. The reported cases (7.3%) were to the National Agency for Food and Drug Administration and Control. The study showed a statistically significant difference between the institution of respondents, knowledge and experience about pharmacovigilance (p < 0.05). CONCLUSION: There is marked under-reporting of cases and a low level of awareness about pharmacovigilance among resident doctors in Edo and Lagos states. This may be what obtains in other states in Nigeria. PMID- 20014359 TI - Use of over-the-counter analgesics and perceived stress among 25-44-year olds. AB - PURPOSE: To examine the association between perceived stress and use of over-the counter analgesics in a representative sample of 25-44-year old adults, and to examine the association across various socio-demographic strata. Furthermore, to examine whether an association between perceived stress and use of over-the counter analgesics attenuates when controlled by potential stress-related pain and discomfort. METHODS: National representative cross-sectional study in Denmark. The study population consisted of men and women aged 25-44 years, n = 4739. The survey was conducted by face-to-face interviews. The outcome measure was use of over-the-counter analgesics (OTCA). The independent variable was perceived stress. Demographic variables and pain and discomfort symptoms were included as covariates. RESULTS: Analyses stratified by socio-demographic factors (gender, education, cohabiting status and whether or not the respondents had children) all showed a significant and graded association between stress and OTCA use. The odds for OTCA use mounted with increasing stress. In analyses adjusted for socio-demographic variables and pain or discomfort the association between stress and OTCA use attenuated somewhat, but remained significant and graded. The crude odds ratio (OR) for OTCA use was 1.36 (1.19-1.55) among participants who sometimes felt stress, and 1.91 (1.58-2.30) among participants who often felt stress, compared to participants without stress. CONCLUSION: There was a significant and graded association between perceived stress and OTCA use. The association was robust across all the examined socio-demographic strata and could not be explained by potential stress-related pain and discomfort. The results indicate that OTCA are used inappropriately to treat feelings of stress. PMID- 20014360 TI - Terms, definitions and measurements to describe the sonographic features of the endometrium and intrauterine lesions: a consensus opinion from the International Endometrial Tumor Analysis (IETA) group. AB - The IETA (International Endometrial Tumor Analysis group) statement is a consensus statement on terms, definitions and measurements that may be used to describe the sonographic features of the endometrium and uterine cavity on gray scale sonography, color flow imaging and sonohysterography. The relationship between the ultrasound features described and the presence or absence of pathology is not known. However, the IETA terms and definitions may form the basis for prospective studies to predict the risk of different endometrial pathologies based on their ultrasound appearance. PMID- 20014361 TI - Iliac crest angle: a novel sonographic parameter for the prediction of Down syndrome risk during the second trimester of pregnancy. AB - OBJECTIVES: To validate a new sonographic technique for the display and measurement of fetal iliac crest angle (ICA), and to determine the relative contribution of standardized fetal pelvic measurements for estimating risk of Down syndrome during the second trimester of pregnancy. METHODS: Three dimensional ultrasonography of the fetal pelvis was performed during a second trimester scan for genetic amniocentesis. A coronal view of the 'virtual pelvis' was obtained by applying a maximum intensity projection algorithm to visualize both iliac wings. Two different ICAs were measured from a coronal projection of the fetal pelvis (ICA-coronal 1 and ICA-coronal 2); the latter approach relied on using ischial tuberosities as reference landmarks. Next, a coronal projection of the fetal pelvis was rotated to demonstrate a rendered view of the axial fetal pelvis. Axial inner (ICA-inner), middle (ICA-middle) and outer (ICA-outer) ICAs were measured. The maximum anterior iliac distance measurement was defined as the widest separation between the most anterior points of the rendered iliac crests. The reproducibility of each ICA method was analyzed using Bland-Altman statistics. Simple and multiple logistic regression analysis determined the relative contribution of each parameter as a prenatal predictor of Down syndrome. RESULTS: Ninety-four normal fetuses and 19 fetuses with Down syndrome were examined. The ICA-middle and ICA-coronal 2 parameters were the most reproducible angle measurements. The mean +/- SD ICA-middle measurement for fetuses with Down syndrome was significantly greater than that for normal subjects (94.5 +/- 9.8 degrees vs. 83.1 +/- 6.5 degrees ; P < 0.001). The mean +/- SD ICA-coronal 2 angle measurement for fetuses with Down syndrome was slightly greater than that for normal subjects (57.9 +/- 5.5 degrees vs. 51.9 +/- 7.1 degrees ; P = 0.0014). A multiple logistic regression model including ICA-middle and ICA-coronal 2 provided a predictive ability of 88.1% based on the area under the receiver operating characteristics curve. This combination had a sensitivity of 94.4% for a false-positive rate of 5% in the detection of Down syndrome. CONCLUSIONS: Standardized iliac crest measurements of the fetal pelvis can be used to identify some fetuses at risk for trisomy 21 during the second trimester of pregnancy. PMID- 20014362 TI - Cognitive profile of Turner syndrome. AB - Turner syndrome (TS) is a relatively common neurogenetic disorder characterized by complete or partial monosomy-X in a phenotypic female. TS is associated with a cognitive profile that typically includes intact intellectual function and verbal abilities with relative weaknesses in visual-spatial, executive, and social cognitive domains. In this report, we review previous and current research related to the cognitive profile of TS. We also discuss how cognitive impairments in this syndrome may reflect integrative rather than modular deficits. For example, the less commonly reported areas of verbal difficulty in TS and certain visual-spatial deficits seem significantly influenced by impairments in executive function and spatially loaded stimuli. We provide a summary of cognitive testing measures used in the assessment of visual-spatial and executive skills, which includes test domain descriptions as well as a comprehensive examination of social cognitive function in TS. This review concludes with a discussion of ecological interpretations regarding the meaning of cognitive deficits in TS at the individual level. PMID- 20014363 TI - Fragile X syndrome -- from genes to cognition. AB - Fragile X syndrome (FXS), a single gene disorder with an expanded CGG allele on the X chromosome, is the most common form of inherited cognitive impairment. The cognitive deficit ranges from mild learning disabilities to severe intellectual disability. The phenotype includes hyperactivity, short attention span, emotional problems including anxiety, social avoidance, poor eye contact, and hyperarousal to sensory stimuli. Imaging studies in FXS have clarified the impact of the FMR1 mutation on brain development and function by documenting structural abnormalities, predominantly in the caudate nucleus and cerebellum, and functional deficits in the caudate, frontal-striatal circuits, and the limbic system. On the basis of current research results, a targeted treatment for FXS will be available in the near future. Currently, a number of psychopharmacological agents are helpful in treating many of the problems in FXS including hyperactivity, attention deficits, anxiety, episodic aggression, and hyperarousal. Although the targeted treatments aim at strengthening synaptic connections, it is essential that these treatments are combined with learning programs that address the cognitive deficits in FXS. PMID- 20014364 TI - X-linked intellectual disability: unique vulnerability of the male genome. AB - X-linked intellectual disability (XLID) accounts for approximately 16% of males with intellectual disability (ID). This is, in part, related to the fact that males have a single X chromosome. Progress in the clinical and molecular characterization of XLID has outpaced progress in the delineation of ID due to genes on the other 22 chromosomes. Almost half of the estimated 200 XLID genes have been identified and another 20% have been regionally mapped. These advances have had immediate benefits for families, allowing for carrier testing, genetic counseling, prenatal diagnosis, and preimplantation genetic diagnosis. Additionally, the combination of clinical delineation with gene identification and the development of gene panels for screening nonsyndromal XLID has been able to limit unproductive laboratory testing. Most importantly for the patients, some of the gene discoveries have pointed to potential strategies for treatment. PMID- 20014365 TI - Introduction: cognitive profiles in sex chromosome disorders. PMID- 20014366 TI - Turner syndrome: neuroimaging findings: structural and functional. AB - Neuroimaging studies of Turner syndrome can advance our understanding of the X chromosome in brain development, and the modulatory influence of endocrine factors. There is increasing evidence from neuroimaging studies that TX individuals have significant differences in the anatomy, function, and metabolism of a number of brain regions; including the parietal lobe; cerebellum, amygdala, hippocampus; and basal ganglia; and perhaps differences in "connectivity" between frontal and parieto-occipital regions. Finally, there is preliminary evidence that genomic imprinting, sex hormones and growth hormone have significant modulatory effects on brain maturation in TS. PMID- 20014367 TI - Social function in multiple X and Y chromosome disorders: XXY, XYY, XXYY, XXXY. AB - Klinefelter syndrome (47,XXY) was initially described in the context of its endocrinologic and physical features; however, subsequent studies have revealed specific impairments in verbal skills and social functioning. Males with sex chromosomal aneuploidies are known to have variability in their developmental profile with the majority presenting with expressive language deficits. As a consequence of language delays, they have an increased likelihood of language based learning disabilities and social-emotional problems that may persist through adulthood. Studies on males with 47,XXY have revealed unique behavioral and social profiles with possible vulnerability to autistic traits. The prevalence of males with more than one extra sex chromosome (e.g., 48,XXYY and 48,XXXY) and an additional Y (e.g., 47,XYY) is less common, but it is important to understand their social functioning as it provides insight into treatment implications. PMID- 20014368 TI - Gene, brain, and behavior relationships in fragile X syndrome: evidence from neuroimaging studies. AB - Fragile X syndrome (FraX) remains the most common inherited cause of intellectual disability and provides a valuable model for studying gene-brain-behavior relationships. Over the past 15 years, structural and functional magnetic resonance imaging studies have emerged with the goal of better understanding the neural pathways contributing to the cognitive and behavioral outcomes seen in individuals with FraX. Specifically, structural MRI studies have established and begun to refine the specific topography of neuroanatomical variation associated with FraX. In addition, functional neuroimaging studies have begun to elucidate the neural underpinnings of many of the unique characteristics of FraX including difficulties with eye gaze, executive functioning, and behavioral inhibition. This review highlights studies with a focus on the relevant gene-brain-behavior connections observed in FraX. The relationship of brain regions and activation patterns to FMRP are discussed as well as the clinical cognitive and behavioral correlates of these neuroimaging findings. PMID- 20014370 TI - Structural and functional neuroimaging in Klinefelter (47,XXY) syndrome: a review of the literature and preliminary results from a functional magnetic resonance imaging study of language. AB - Klinefelter (47,XXY) syndrome (KS), the most common form of sex-chromosomal aneuploidy, is characterized by physical, endocrinologic, and reproductive abnormalities. Individuals with KS also exhibit a cognitive/behavioral phenotype characterized by language and language-based learning disabilities and executive and attentional dysfunction in the setting of normal general intelligence. The underlying neurobiologic mechanisms are just now beginning to be elucidated through structural and functional neuroimaging. Here, we review the literature of structural and functional neural findings in KS identified by neuroimaging and present preliminary results from a functional magnetic resonance imaging study examining brain activity during a verb generation task in KS. PMID- 20014369 TI - The cognitive phenotype in Klinefelter syndrome: a review of the literature including genetic and hormonal factors. AB - Klinefelter syndrome (KS) or 47,XXY occurs in approximately 1 in 650 males. Individuals with KS often present with physical characteristics including tall stature, hypogonadism, and fertility problems. In addition to medical findings, the presence of the extra X chromosome can lead to characteristic cognitive and language deficits of varying severity. While a small, but significant downward shift in mean overall IQ has been reported, the general cognitive abilities of patients with KS are not typically in the intellectual disability range. Most studies support that males with KS have an increased risk of language disorders and reading disabilities. Results of other studies investigating the relationship between verbal and nonverbal/spatial cognitive abilities have been mixed, with differing results based on the age and ascertainment method of the cohort studied. Executive function deficits have been identified in children and adults with KS, however, the research in this area is limited and further investigation of the neuropsychological profile is needed. In this article, we review the strengths and weaknesses of previous cognitive and neuropsychological studies in males with KS in childhood and adulthood, provide historical perspective of these studies, and review what is known about how hormonal and genetic factors influence cognitive features in 47,XXY/KS. PMID- 20014371 TI - An extra X or Y chromosome: contrasting the cognitive and motor phenotypes in childhood in boys with 47,XYY syndrome or 47,XXY Klinefelter syndrome. AB - OBJECTIVE: The goal of this study was to contrast the cognitive phenotypes in boys with 47,XYY (XYY) karyotype and boys with 47,XXY karyotype [Klinefelter syndrome, (KS)], who share an extra copy of the X-Y pseudoautosomal region but differ in their dosage of strictly sex-linked genes. METHODS: Neuropsychological evaluation of general cognitive ability, language, memory, attention, visual spatial abilities, visual-motor skills, and motor function. RESULTS: Study cohort: 21 boys with 47,XYY and 93 boys with 47,XXY (KS), age 4-17 years, and 36 age-matched control boys. Both the XYY and KS groups performed less well, on average, than the controls on tests of general cognitive ability, achievement, language, verbal memory, some aspects of attention, and executive function, and motor function. The boys with XYY on average had more severe and pervasive language impairment, at both simple and complex levels, and the boys with KS on average had greater motor impairment in gross motor function and coordination, especially in running speed and agility. CONCLUSIONS: The results from these large XYY and KS cohorts have important neurocognitive and educational implications. From the neurocognitive standpoint, the presenting findings afford an opportunity to gain insights into brain development in boys with XYY and those with KS. From the educational standpoint, it is critical that boys with XYY or KS receive appropriate educational interventions that target their specific learning challenges. These findings also provide important information for counseling clinicians and families about these disorders. PMID- 20014372 TI - Effects of sex chromosome aneuploidies on brain development: evidence from neuroimaging studies. AB - Variation in the number of sex chromosomes is a relatively common genetic condition, affecting as many as 1/400 individuals. The sex chromosome aneuploidies (SCAs) are associated with characteristic behavioral and cognitive phenotypes, although the degree to which specific individuals are affected can fall within a wide range. Understanding the effects of different dosages of sex chromosome genes on brain development may help to understand the basis for functional differences in affected individuals. It may also be informative regarding how sex chromosomes contribute to typical sexual differentiation. Studies of 47,XXY males make up the bulk of the current literature of neuroimaging studies in individuals with supernumerary sex chromosomes, with a few small studies or case reports of the other SCAs. Findings in 47,XXY males typically include decreased gray and white matter volumes, with most pronounced effects in the frontal and temporal lobes. Functional studies have shown evidence of decreased lateralization. Although the hypogonadism typically found in 47,XXY males may contribute to the decreased brain volume, the observation that 47,XXX females also show decreased brain volume in the presence of normal pubertal maturation suggests a possible direct dosage effect of X chromosome genes. Additional X chromosomes, such as in 49,XXXXY males, are associated with more markedly decreased brain volume and increased incidence of white matter hyperintensities. The limited data regarding effects of having two Y chromosomes (47,XYY) do not find significant differences in brain volume, although there are some reports of increased head size. PMID- 20014374 TI - Defying ring strain: new approaches to cyclopropanes. PMID- 20014373 TI - Treatments for fragile X syndrome: a closer look at the data. AB - Research into the determinants and developmental course of fragile X syndrome (FXS) has made remarkable progress over the last 25 years. However, treatments to ameliorate the symptoms of FXS have been less forthcoming. While there is optimism in the field that the pace of intervention research is quickening, there has been a bias toward psychopharmacological approaches to treatment. A closer look at the data from those investigations reveals a paucity of evidence that medications can improve intellectual and adaptive functioning in FXS, or decrease associated behavioral and/or emotional issues. Work in other related disorders (e.g., autism) has shown that dramatic improvements in intellectual and adaptive functioning, as well as behavioral and emotional problems, can occur if intensive behavioral treatment is begun early in the child's life. It is hoped that future research efforts will evaluate these intensive early intervention strategies in children with FXS, perhaps in combination with pharmacological approaches. PMID- 20014375 TI - Efficient access to extended Yagupolskii-Umemoto-type reagents: triflic acid catalyzed intramolecular cyclization of ortho-ethynylaryltrifluoromethylsulfanes. PMID- 20014376 TI - Gold(I)-catalyzed enantioselective synthesis of pyrazolidines, isoxazolidines, and tetrahydrooxazines. PMID- 20014377 TI - Catalytic enantio- and diastereoselective alkylations with cyclic sulfamidates. PMID- 20014378 TI - Desymmetrization of meso-2-alkene-1,4-diol derivatives through copper(I) catalyzed asymmetric boryl substitution and stereoselective allylation of aldehydes. PMID- 20014379 TI - Dynamics of hippocampal spatial representation in echolocating bats. AB - The "place fields" of hippocampal pyramidal neurons are not static. For example, upon a contextual change in the environment, place fields may "remap" within typical timescales of ~ 1 min. A few studies have shown more rapid dynamics in hippocampal activity, linked to internal processes, such as switches between spatial reference frames or changes within the theta cycle. However, little is known about rapid hippocampal place field dynamics in response to external, sensory stimuli. Here, we studied this question in big brown bats, echolocating mammals in which we can readily measure rapid changes in sensory dynamics (sonar signals), as well as rapid behavioral switches between distal and proximal exploratory modes. First, we show that place field size was modulated by the availability of sensory information, on a timescale of ~ 300 ms: Bat hippocampal place fields were smallest immediately after an echolocation call, but place fields "diffused" with the passage of time after the call, when echo information was no longer arriving. Second, we show rapid modulation of hippocampal place fields as the animal switched between two exploratory modes. Third, we compared place fields and spatial view fields of individual neurons and found that place tuning was much more pronounced than spatial view tuning. In addition, dynamic fluctuations in spatial view tuning were stronger than fluctuations in place tuning. Taken together, these results suggest that spatial representation in mammalian hippocampus can be very rapidly modulated by external sensory and behavioral events. PMID- 20014380 TI - Long-term visuospatial retention unaffected by fornix transection. AB - As part of an earlier experiment (Kwok and Buckley, 2009), six macaque monkeys (three with fornix transection and three unoperated controls) were trained postoperatively to discriminate a total of 104 new concurrent visuospatial conditional problems to criterion. Our experiment measured and compared long-term retention of these problems with two separate one-trial postoperative retention tests administered 3 and 15 months, respectively, after acquisition. All animals showed some degree of forgetting of these problems but all remembered above chance levels, even after 15 months. The amount forgotten by each group did not differ significantly at either time point. These results show that long-term retention of visuospatial information is independent of the fornix. Similarities in resistance to forgetting are drawn between fornix-transected macaques and patients with amnesia and the implications for clinical rehabilitation are discussed. PMID- 20014382 TI - Expression of fatty acid-binding proteins in adult hippocampal neurogenic niche of postischemic monkeys. AB - Intracellular fatty acid (FA) chaperones known as FA-binding proteins (FABPs) are a group of molecules known to participate in cellular metabolic processes such as lipid storage, membrane synthesis, and beta-oxidation or to coordinate transcriptional programs. However, their role in adult neurogenesis still remains obscure. The FABPs expressed in the central nervous system (CNS) are heart-type (FABP3), epidermal-type (FABP5), and brain-type (FABP7). These three FABPs possess a differential affinity for polyunsaturated fatty acids (PUFAs). Recently, we reported that GPR40, a receptor for free FAs and particularly for PUFAs, is expressed in the CNS of adult monkeys and upregulated after transient global brain ischemia in the hippocampal subgranular zone (SGZ), a neurogenic niche in adulthood. The SGZ showed a peak proliferation of progenitor cells and maximal expression of GPR40 during the second week after ischemia. As both FABPs and GPR40 might be closely related to the adult neurogenesis, here, we studied the expression of FABP 3, 5, and 7 in the SGZ, comparing normal and postischemic adult monkeys. Immunoblotting revealed that FABP5 and FABP7, but not FABP3, were significantly increased on day 15 after ischemia when compared with the nonischemic control. Immunohistochemistry showed that FABP5 was almost undetectable in the control SGZ but was abundant on day 15 after ischemia. FABP 3, 5, and 7 were expressed in S-100beta-positive astrocytes and nestin-positive neural progenitors. However, only FABP 5 and 7 were found in bromodeoxyuridine (BrdU)-positive newly generated cells. FABPs were most frequently coexpressed with the S-100beta-positive astrocytes, whereas betaIII-tubulin-or polysialylated neural cell-adhesion molecule (PSA-NCAM)-positive newborn neurons in the vicinity of the astrocytes expressed none of the three FABPs. These results support a role of astrocyte- and/or neural progenitor-derived FABPs as components of the molecular machine regulating the progenitor cell niche in the adult primate brain. PMID- 20014383 TI - Developmental regulation of gene expression and astrocytic processes may explain selective hippocampal vulnerability. AB - The hippocampus plays a central role in the brain network that is essential for memory function. Paradoxically, the hippocampus is also the brain structure that is most sensitive to hypoxic-ischemic episodes. Here, we show that the expression of genes associated with glycolysis and glutamate metabolism in astrocytes and the coverage of excitatory synapses by astrocytic processes undergo significant decreases in the CA1 field of the monkey hippocampus during postnatal development. Given the established role of astrocytes in the regulation of glutamate concentration in the synaptic cleft, our findings suggest that a developmental decrease in astrocytic processes could underlie the selective vulnerability of CA1 during hypoxic-ischemic episodes in adulthood, its decreased susceptibility to febrile seizures with age, as well as contribute to the emergence of selective, adultlike memory function. PMID- 20014381 TI - Developmental profiling of postnatal dentate gyrus progenitors provides evidence for dynamic cell-autonomous regulation. AB - The dentate gyrus of the hippocampus is one of the most prominent regions in the postnatal mammalian brain where neurogenesis continues throughout life. There is tremendous speculation regarding the potential implications of adult hippocampal neurogenesis, though it remains unclear to what extent this ability becomes attenuated during normal aging, and what genetic changes in the progenitor population ensue over time. Using defined elements of the nestin promoter, we developed a transgenic mouse that reliably labels neural stem and early progenitors with green fluorescent protein (GFP). Using a combination of immunohistochemical and flow cytometry techniques, we characterized the progenitor cells within the dentate gyrus and created a developmental profile from postnatal day 7 (P7) until 6 months of age. In addition, we demonstrate that the proliferative potential of these progenitors is controlled at least in part by cell-autonomous cues. Finally, to identify what may underlie these differences, we performed stem cell-specific microarrays on GFP-expressing sorted cells from isolated P7 and postnatal day 28 (P28) dentate gyrus. We identified several differentially expressed genes that may underlie the functional differences that we observe in neurosphere assays from sorted cells and differentiation assays at these different ages. These data suggest that neural progenitors from the dentate gyrus are differentially regulated by cell autonomous factors that change over time. PMID- 20014384 TI - Deletion of the L-type calcium channel Ca(V) 1.3 but not Ca(V) 1.2 results in a diminished sAHP in mouse CA1 pyramidal neurons. AB - Trains of action potentials in CA1 pyramidal neurons are followed by a prolonged calcium-dependent postburst afterhyperpolarization (AHP) that serves to limit further firing to a sustained depolarizing input. A reduction in the AHP accompanies acquisition of several types of learning and increases in the AHP are correlated with age-related cognitive impairment. The AHP develops primarily as the result of activation of outward calcium-activated potassium currents; however, the precise source of calcium for activation of the AHP remains unclear. There is substantial experimental evidence suggesting that calcium influx via voltage-gated L-type calcium channels (L-VGCCs) contributes to the generation of the AHP. Two L-VGCC subtypes are predominately expressed in the hippocampus, Ca(V) 1.2 and Ca(V) 1.3; however, it is not known which L-VGCC subtype is involved in generation of the AHP. This ambiguity is due in large part to the fact that at present there are no subunit-specific agonists or antagonists. Therefore, using mice in which the gene encoding Ca(V) 1.2 or Ca(V) 1.3 was deleted, we sought to determine the impact of alterations in levels of these two L-VCGG subtypes on neuronal excitability. No differences in any AHP measure were seen between neurons from Ca(V) 1.2 knockout mice and controls. However, the total area of the AHP was significantly smaller in neurons from Ca(V) 1.3 knockout mice as compared with neurons from wild-type controls. A significant reduction in the amplitude of the AHP was also seen at the 1 s time point in neurons from Ca(V) 1.3 knockout mice as compared with those from controls. Reductions in both the area and 1 s amplitude suggest the involvement of calcium influx via Ca(V) 1.3 in the slow AHP (sAHP). Thus, the results of our study demonstrate that deletion of Ca(V) 1.3, but not Ca(V) 1.2, significantly impacts the generation of the sAHP. PMID- 20014385 TI - Altered patterning of dentate granule cell mossy fiber inputs onto CA3 pyramidal cells in limbic epilepsy. AB - Impaired gating by hippocampal dentate granule cells may promote the development of limbic epilepsy by facilitating seizure spread through the hippocampal trisynaptic circuit. The second synapse in this circuit, the dentate granule cell?CA3 pyramidal cell connection, may be of particular importance because pathological changes occurring within the dentate likely exert their principal effect on downstream CA3 pyramids. Here, we utilized GFP-expressing mice and immunolabeling for the zinc transporter ZnT-3 to reveal the pre- and postsynaptic components of granule cell?CA3 pyramidal cell synapses following pilocarpine epileptogenesis. Confocal analyses of these terminals revealed that while granule cell presynaptic giant boutons increased in size and complexity 1 month after status epilepticus, individual thorns making up the postsynaptic thorny excrescences of the CA3 pyramidal cells were reduced in number. This reduction, however, was transient, and 3 months after status, thorn density recovered. This recovery was accompanied by a significant change in the distribution of thorns along pyramidal cells dendrites. While thorns in control animals tended to be tightly clustered, thorns in epileptic animals were more evenly distributed. Computational modeling of thorn distributions predicted an increase in the number of boutons required to cover equivalent numbers of thorns in epileptic vs. control mice. Confirming this prediction, ZnT-3 labeling of presynaptic giant boutons apposed to GFP-expressing thorns revealed a near doubling in bouton density, while the number of individual thorns per bouton was reduced by half. Together, these data provide clear evidence of novel plastic changes occurring within the epileptic hippocampus. PMID- 20014387 TI - Hippocampal signals for strong memory when associative memory is available and when it is not. AB - The paired-associate task has been used with functional magnetic resonance imaging (fMRI) in studies that assessed the role of the medial temporal lobe (MTL) subserving recollection and familiarity.Some researchers have interpreted their results to mean that the hippocampus selectively subserves recollection and not familiarity[cf., Eichenbaum et al., (2007) Annu Rev Neurosci 30:123-152]. Yet many of these results confound recollection and familiarity with strong and weak memories, and it is not clear whether the conclusions represent differences between memory processes or memory strength. In the current study, participants were scanned with fMRI during retrieval in a paired-associate task, and a new approach separated the analysis of memory strength from the analysis of memory processes. The data were sorted by confidence level in an old/new task, and the high-confidence responses were compared in categories when associative memory was highly accurate and when it was not available. The results show that high confidence memory produced increased activity in the hippocampus,relative to the level for forgotten pairs, both when associative memory was available and when it was not. Two interpretations are discussed for the behavioral results for when associative memory was not available: one account based on familiarity and the other account based on noncriterial recollection. The conclusion is that recognition of the word-pairs was based on familiarity when associative memory was not available. Together with the fMRI results that activity in two regions associated with cognitive control (left ventrolateral prefrontal cortex and left inferior parietal lobule) was greater when responses were based on associative memory than when based on familiarity, the findings suggest that the hippocampus supports strong memory and that cortical regions make an additional contribution to recollection. PMID- 20014386 TI - Phosphacan and receptor protein tyrosine phosphatase beta expression mediates deafferentation-induced synaptogenesis. AB - This study documents the spatial and temporal expression of three structurally related chondroitin sulfated proteoglycans (CSPGs) during synaptic regeneration induced by brain injury. Using the unilateral entorhinal cortex (EC) lesion model of adaptive synaptogenesis, we documented mRNA and protein profiles of phosphacan and its two splice variants, full length receptor protein tyrosine phosphatase beta (RPTPbeta) and the short transmembrane receptor form (sRPTPbeta), at 2, 7, and 15 days postlesion. We report that whole hippocampal sRPTPbeta protein and mRNA are persistently elevated over the first two weeks after UEC. As predicted, this transmembrane family member was localized adjacent to synaptic sites in the deafferented neuropil and showed increased distribution over that zone following lesion. By contrast, whole hippocampal phosphacan protein was not elevated with deafferentation; however, its mRNA was increased during the period of sprouting and synapse formation (7d). When the zone of synaptic reorganization was sampled using molecular layer/granule cell (ML/GCL) enriched dissections, we observed an increase in phosphacan protein at 7d, concurrent with the observed hippocampal mRNA elevation. Immunohistochemistry also showed a shift in phosphacan distribution from granule cell bodies to the deafferented ML at 2 and 7d postlesion. Phosphacan and sRPTPbeta were not colocalized with glial fibrillary acid protein (GFAP), suggesting that reactive astrocytes were not a major source of either proteoglycan. While transcript for the developmentally prominent full length RPTPbeta was also increased at 2 and 15d, its protein was not detected in our adult samples. These results indicate that phosphacan and RPTPbeta splice variants participate in both the acute degenerative and long-term regenerative phases of reactive synaptogenesis. These results suggest that increase in the transmembrane sRPTPbeta tyrosine phosphatase activity is critical to this plasticity, and that local elevation of extracellular phosphacan influences dendritic organization during synaptogenesis. PMID- 20014388 TI - A case of right low partial jugular vein duplication. PMID- 20014389 TI - The anatomical basis of venographic filling defects of the transverse sinus. AB - Obstruction of the intracranial dural venous sinuses would result in an increase in intracranial dural venous pressure. This intracranial hypertension is not only the result of poor cerebral venous drainage but also life threatening. The aim of this study was to identify the structures, which may show signs of potential venographic filling defect qualities, including trabeculae/septa (also described as "fibrous bands") and arachnoid granulations, which ultimately can lead to increased intracranial dural sinus venous pressure. A total of 102 cadavers and living patients were used for the study. Fifty-three percent of the subjects presented with structures in their transverse sinuses that could be potential venous filling defects. Thirty percent of the subjects presented with arachnoid granulations in the right transverse sinus, which were found to be significantly dominant (Chi-square; p < 0.05). The study also revealed the presence of 1 to 5 septa in 29.4% of the subjects. The septa were found to be more dominant in the central (30%) and lateral (22%) thirds of the right transverse sinuses, while the central third of the left transverse sinus proved to be the least dominant occurring site (8%). In general, the right transverse sinus is highly more significantly dominant in septal occurrence (Chi-square; p < 0.01) than the left transverse sinus. We conclude from the statistical evidence that the right transverse sinus demonstrates significantly more potential venographic filling defects than the left sinus and submit that this information may assist in management options for patients diagnosed with idiopathic intracranial hypertension as well as direct future research. PMID- 20014390 TI - Anatomical variations in the level of origin of the sigmoid colon from the descending colon and the attachment of the sigmoid mesocolon. AB - The origin of the sigmoid colon is considered constant as is the V-shaped attachment of the sigmoid mesocolon attachment. This study was undertaken to establish anatomical variations in the level of origin of the sigmoid colon (590 autopsies; 403 Africans, 91 Indians, and 96 Whites), and the shape of the attachment of the sigmoid mesocolon (211 autopsies, 127 Africans, 47 Indians, and 37 Whites) in different population groups. The low-level origin was significantly less common among Africans compared with the other population groups (P = 0.003) and the high-level origin was significantly more common in Africans (P = 0.003). A midlevel origin was similar in all three groups. The shape of the mesocolon attachment was either straight (94), inverted U-shaped (79), or inverted V-shaped (38). The straight shape was more common in Whites (Whites vs. African and Indian P = 0.003), and the U-shape more common in Africans (African vs. Whites P = 0.042). The distribution of the V-shape was similar. There are anatomical variations in the level of origin of the sigmoid colon from the descending colon as well as in the shape of the attachment of its mesocolon. These variations are population based. PMID- 20014391 TI - About the arterial anatomy of the Achilles tendon (tendo calcaneus). PMID- 20014392 TI - Possible nociceptive structures in the sacroiliac joint cartilage: An immunohistochemical study. AB - The sacroiliac joint (SI joint) is a known source of low back pain. In the absence of validated physical signs and imaging studies, the diagnosis of SI joint pain can be secured by positive response to SI joint intra-articular infiltration with local anesthetics. The current anatomical and histological knowledge concerning intra-articular structures of the sacroiliac joint is insufficient to explain the efficacy of this infiltration. Consequently, this study was undertaken to detect the intra-articular presence of substance P and calcitonin gene-related peptide (CGRP) positive nerve fibers, providing indirect evidence of nociceptive innervation of the SI joint. Free-floating sections, obtained from iliac and sacral cartilage and subchondral bone of the SI joint and adjacent ligamentous tissue, of 10 human cadavers were studied immunohistochemically. Tissue of nine human cadavers showed the presence of substance P and CGRP immunoreactivity in the superficial layer of sacral and iliac cartilage, and the surrounding ligamentous structures. Subchondral bone reacted weakly to the antisera used. These findings support the view that the SI joint may be capable of intra-articular nociception and may explain the positive response to the intra-articular deposition of local anesthetic. PMID- 20014393 TI - Response to the commentary on "Anatomy in the Third Reich: An outline" by G. Aumuller and R. Hildebrand. PMID- 20014395 TI - The spinoglenoid foramen. PMID- 20014396 TI - Imaging study of the long extracranial extension of the inferior petrosal sinus with MSCT. AB - The aim of this study was to explore the anatomic route of Long-Inferior Petrosal Sinuses (IPS) with multi-slice spiral computed tomography, and to provide referenced evidence for the interventional preoperative evaluation for the diagnosis and treatment of skull base and sellar lesions. The route of Long-IPS and its confluence with the internal jugular vein (IJV) and the connection level of 12 IPS were shown with multi-planar reconstruction and curved multi-planar reconstruction, and the IPS length was determined. Combining the results of continuous multi-slice scanning, the diameters of the IPS at the initial segment in the jugular foreman and middle segment and the confluence segment of the IPS IJV junction level were determined. The mean length of the Long-IPS was 66.2 +/- 17.5 mm, and the length was over 60 mm on eight sides and its peak value 100 mm. The mean diameters of the IPS were 2.4 mm +/- 0.7 mm, 2.1 mm +/- 0.4 mm, and 2.1 mm +/- 0.5 mm at the initial, middle, and confluent segments, respectively. Their diameters were equal to or greater than 2 mm at the connection level on eight sides. Furthermore, the diameter was greater than 1.6 mm at the middle and initial segments. The Long-IPS might be used as a route to the intra-cranial IPS. MSCT is helpful for showing the route and variation of the IPS and could be an effective method for preoperative evaluation of the IPS. PMID- 20014397 TI - Communications between the palmar digital branches of the median and ulnar nerves: A study in human fetuses and a review of the literature. AB - In this study, median nerves (MNs) and ulnar nerves (UNs) were dissected in 200 palmar sides of hands (left and right) of 100 (50 male, 50 female) spontaneously aborted fetuses with no detectable malformations. The fetuses, whose gestational ages ranged from 13 to 40 weeks, were dissected under an operating microscope. The MN divided first into a lateral ramus and a medial ramus and then formed a common digital nerve. The first common digital nerve trifurcated in all of the studied cases. The branching patterns were classified into two types (Type 1 and Type 2) based on the relationship with the flexor retinaculum (behind/distal of it). A communication branch between the UNs and MNs in the palmar surface of the hand was found in 59 hands (29.5%). The proper palmar digital nerves were numbered from p1 to p10, starting from the radial half of the thumb to the ulnar half of the little finger, and these nerves exhibited six types of variations. The present data obtained from human fetuses will aid in elucidating the developmental anatomy of the nervous system and provide hand surgeons with a more complete anatomical picture to help them to avoid iatrogenic injuries. PMID- 20014400 TI - Prognostic significance of grading in lung adenocarcinoma. AB - BACKGROUND: Although grading has prognostic significance for many tumor types, a prognostically significant grading system for lung adenocarcinoma has not yet been established. The aim of this study was to evaluate histologic characteristics included in tumor grading systems, establish optimal cutoff values that have the strongest association with overall survival, and develop a grading system incorporating the histopathologic characteristics that the authors found to have prognostic significance in patients with lung adenocarcinoma. METHODS: The authors studied lung adenocarcinomas from 85 consecutive patients, and evaluated the percentage of solid pattern (as a reflection of tumor architecture), the degree of cytologic atypia, and the mitotic count. RESULTS: In univariate analysis, overall survival was associated significantly with sex (P = .045), age (P = .0008), tumor status (P < .0001), lymph node status (P = .02), solid pattern (P = .046), and cytologic atypia (P = .01), but not with mitotic count (P = .26). On the basis of optimal cutoff values, the authors found that a solid pattern > or = 90% and severe cytologic atypia were the best discriminators of worse outcome. A grading score, computed as the sum of the architecture score and cytologic atypia score (2 = well differentiated, 3 = moderately differentiated, 4 = poorly differentiated), was a significant predictor of overall survival in univariate analysis (median overall survival times, 72.4, 39.5, and 8.7 months for well, moderately, and poorly differentiated adenocarcinoma, respectively; P = .0001). Moreover, grading was an independent predictor of survival in multivariate analysis (P = .002). CONCLUSIONS: The authors describe a grading system that incorporates the percentage of solid pattern and degree of the cytologic atypia that is an independent predictor of survival in patients with lung adenocarcinoma. PMID- 20014399 TI - Radiotherapy for patients with the human immunodeficiency virus: are special precautions necessary? AB - Shortly after the onset of the acquired immunodeficiency syndrome (AIDS) epidemic in the 1980s, reports of radiation-associated toxicity in patients with the human immunodeficiency virus (HIV) and AIDS began to appear in the medical literature. Although the majority of reports have focused on AIDS-defining malignancies such as Kaposi sarcoma, greater-than-expected toxicity after a course of radiotherapy or chemoradiotherapy has also been documented in cancers not generally classified as being related to HIV. With improved antiretroviral therapies, HIV patients are living longer and have the potential to develop a variety of HIV-associated and nonassociated malignancies that require treatment, including radiotherapy. This review reports the published data regarding the interactions of HIV, AIDS, and antiretroviral therapy with radiotherapy and implications for the management of malignancies in patients with HIV. PMID- 20014401 TI - The surgical anatomy of the radial nerve and the triceps aponeurosis. AB - The radial nerve passes around the posterior aspect of the humerus where it is prone to injury in both humeral fractures and surgical exploration of this region. We examined 55 cadaveric limbs to determine whether the exact position of the radial nerve could be reliably predicted on the basis of superficial anatomical markings. We found that when there is considerable variability in the position of the nerve in relation to the lateral epicondyle, the nerve consistently passed adjacent to the lateral border of the triceps aponeurosis at a distance of 22-27 (+/-2) mm. It was never found to be closer than 13 (+/-1) mm to the aponeurosis. The lateral border of the triceps aponeurosis is easy to identify and our findings may help avoid iatrogenic injury to the radial nerve during exploration. PMID- 20014402 TI - The iguana/DZIP1 protein is a novel component of the ciliogenic pathway essential for axonemal biogenesis. AB - Cilia play important roles in many developmental and physiological processes. However, the genetic and cell biological control of ciliogenesis remains poorly understood. Here, we show that the zebrafish iguana gene is required for differentiation of primary cilia. iguana encodes a zinc finger and coiled-coil containing protein, previously implicated in Hedgehog signaling. We now argue that aberrant Hedgehog activity in iguana -deficient zebrafish arises from their profound lack of primary cilia. By contrast, the requirement of iguana for motile cilia formation is less obligatory. In the absence of iguana function, basal bodies can migrate to the cell surface and appear to engage with the apical membrane. However, formation of ciliary pits and axonemal outgrowth is completely inhibited. Iguana localizes to the base of primary and motile cilia, in the immediate vicinity or closely associated with the basal bodies. These findings identify the Iguana protein as a novel and critical component of ciliogenesis. PMID- 20014403 TI - Knockdown of the neuronal nitric oxide synthase gene retard the development of the cerebellar granule neurons in vitro. AB - The role of endogenous neuronal nitric oxide synthase (nNOS) gene in the development of cerebellar granule neurons (CGNs) is conflicting. Here, we tested the effect of antisense oligos (AS-ODN) on the endogenous nNOS gene and the development of the CGNs in vitro. The expression of nNOS increased in a development-dependent pattern both in terms of mRNA and protein. AS-ODN down regulated nNOS gene, but in a posttranscriptional manner. Knockdown of nNOS protein decreased the viability of the CGNs from 7 to 13 days in culture (DIC). This activity of AS-ODN was mimicked by nNOS inhibitor I. The antagonist (nNOSi, MK-801, or ODQ) -induced decrease of cell viability was normalized by the provision of the sodium nitroprusside, an NO donor. This study provides direct evidence that endogenous nNOS, mainly by means of its principal product NO, plays an active role in sustaining the survival of developing CGNs at transition from differentiation to maturation. PMID- 20014404 TI - The zebrafish homeobox gene irxl1 is required for brain and pharyngeal arch morphogenesis. AB - Iroquois homeobox-like 1 (irxl1) is a novel member of the TALE superfamily of homeobox genes that is most closely related to the Iroquois class. We have identified the zebrafish irxl1 gene and characterized its structure. The protein contains a homeodomain that shares 100% sequence identity with other vertebrate orthologs. During embryogenesis, irxl1 is expressed from 18 hours postfertilization onward and prominent expression is detected in the pharyngeal arches. Knockdown of irxl1 by morpholinos results in malformed brain and arch structures, which can be partially rescued by cRNA injection. The heads of the morphants become small and flat, and extensions along the anterior posterior/dorso-ventral axes are reduced without affecting regional specification. Loss of irxl1 function also causes deficit in neural crest cells which consequently results in partial loss of craniofacial muscles and severe deformation of arch cartilages. These observations suggest that irxl1 may regulate factors involved in brain and pharyngeal arch development. PMID- 20014405 TI - DNA-adducts in subjects exposed to urban air pollution by benzene and polycyclic aromatic hydrocarbons (PAHs) in Cotonou, Benin. AB - Air pollution effect on humans represents a major public health problem. Exposure to genotoxic compounds in the ambient air is evaluated using different biomarkers. In the present study we assessed DNA-adducts levels in apparently healthy people living and working in the city of Cotonou (Benin) in which exposure to air pollutants such as benzene and polycyclic aromatic hydrocarbons (PAHs) mainly benzo(a)pyrene has been evidenced. Rural inhabitants were enrolled as control group. Taxi-motorbike drivers, street food vendors, and gasoline salesmen were recruited in Cotonou whereas suburban residents were recruited in Godomey, 12 km from Cotonou. We found that taxi-motorbike drivers, roadside residents, street vendors, taxi-motor-bike drivers and gasoline sellers had significantly higher levels of DNA-adducts than suburban and village inhabitants (P < 0.001; post hoc, LSD). Means values were 24.6 +/- 6.4, 23.78 +/- 6.9, 34.7 +/- 9.8, and 37.2 +/- 8.1 in the exposed groups versus 2.1 +/- 0.6 and 3.1 +/- 0.8 adducts/10(8) nucleotides, in the two control groups, respectively. We did not find any significant difference within the high exposure groups and inside low exposure subgroups (namely suburban residents and villagers) because the mean individual exposure values to both PAHs and benzene were similar among subjects exposed in the city of Cotonou and those in suburban and village areas. However, there is significant interindividual variations in adducts levels that may reflect variation of genetic susceptibility factors. Ranges of adduct level/10(8) nucleotides were: 1-69, 1-76, 3-169, 4-124, 0-9, 0-8 adducts/10(8) for taxi motorbike drivers, roadside residents, street vendors, gasoline sellers, suburban and village inhabitants, respectively. Our study demonstrated a clear-cut elevated level of DNA adducts in city residents than in none exposed people (or very low exposure levels people) and designate these city residents groups as people at risks for the chronic diseases possibly caused by benzene and PAHs. PMID- 20014406 TI - POSH misexpression induces caspase-dependent cell death in Drosophila. AB - POSH (Plenty of SH3 domains) is a scaffold for signaling proteins regulating cell survival. Specifically, POSH promotes assembly of a complex including Rac GTPase, mixed lineage kinase (MLK), MKK7, and Jun kinase (JNK). In Drosophila, genetic analysis implicated POSH in Tak1-dependent innate immune response, in part through regulation of JNK signaling. Homologs of the POSH signaling complex components, MLK and MKK7, are essential in Drosophila embryonic dorsal closure. Using a gain-of-function approach, we tested whether POSH plays a role in this process. Ectopic expression of POSH in the embryo causes dorsal closure defects due to apoptosis of the amnioserosa, but ectodermal JNK signaling is normal. Phenotypic consequences of POSH expression were found to be dependent on Drosophila Nc, the caspase-9 homolog, but only partially on Tak1 and not at all on Slpr and Hep. These results suggest that POSH may use different signaling complexes to promote cell death in distinct contexts. PMID- 20014407 TI - Vitamin C coadministration augments bisphenol A, nonylphenol, and octylphenol induced oxidative damage on kidney of rats. AB - The aim of this study was to investigate whether bisphenol A (BPA), nonylphenol (NP), and octylphenol (OP) induce oxidative stress on the kidney tissue of male rats and whether coadministration of vitamin C, an antioxidant, can prevent any possible oxidative stress. The Wistar male rats were divided into seven groups, including control, BPA, NP, OP, BPA+C, NP + C, OP +C. BPA, NP, and OP (25 mg/kg/day) was administered alone; vitamin C (60 mg/kg/day) was administered along with BPA, OP, and NP to the rats for 50 days. There was a decrease in serum concentration of blood urea nitrogen (BUN) in NP and OP groups compared with control group. Vitamin C coadministration with BPA, NP, and OP did not produce significant increase in BUN concentration in BPA +C, NP+ C, and OP + C group as compared with BPA, NP, and OP groups, respectively. The lowest serum creatinine activity and the highest lactate dehydrogenase (LDH) activity was present in kidney of BPA+C, NP+C and OP+C groups compared with BPA, NP, and OP groups. The malondialdehyde (MDA) levels were significantly higher while glutathione (GSH) levels were lower in treatment groups than controls. Furthermore, an increase was observed in MDA levels whereas a decrease was observed in GSH levels in BPA+ C, NP + C, and OP+ C groups compared with BPA, NP, and OP groups, respectively. These finding are in accordance with immunohistochemical staining of MDA and GSH. Histopathological examination of the kidneys of rats in BPA, OP, NP, BPA+ C, NP + C, and OP+ C groups revealed necrotic lesions, congestion, and mononuclear cell infiltration. In conclusion BPA, NP, and OP might induce oxidative damage in kidney of rats. In addition, coadministration of vitamin C with BPA, NP, and OP to male rats augments this damage in the kidney of male rats. PMID- 20014408 TI - EphA4 is necessary for spatially selective peripheral somatosensory topography. AB - Somatosensation is the primary sensory modality employed by rodents in navigating their environments, and mystacial vibrissae on the snout are the primary conveyors of this information to the murine brain. The layout of vibrissae is spatially stereotyped and topographic connections faithfully maintain this layout throughout the neuraxis. Several factors have been shown to influence general vibrissal innervation by trigeminal neurons. Here, the role of a cell surface receptor, EphA4, in directing position-dependent vibrissal innervation is examined. EphA4 is expressed in the ventral region of the presumptive whisker pad and EphA4(-/-) mice lack the ventroposterior-most vibrissae. Analyses reveal that ventral trigeminal axons are abnormal, failing to innervate emerging vibrissae, and resulting in the absence of a select group of vibrissae in EphA4(-/-) mice. EphA4's selective effect on a subset of whiskers implicates cell-based signaling in the establishment of position-dependent connectivity and topography in the peripheral somatosensory system. PMID- 20014409 TI - A study of the subchronic effects of arsenic exposure on the liver tissues of Labeo rohita using Fourier transform infrared technique. AB - In this work, an attempt has been made to study the subchronic effects of arsenic exposure on the biochemical composition; mainly proteins of the liver tissues of Labeo rohita fingerlings by using Fourier transform infrared (FT-IR) spectroscopic technique. The study was carried out using a Perkin Elmer-Spectrum Rx1 spectrometer. Because of arsenic exposure, significant reductions in the intensity as well as area of amide bands have been observed in the liver tissues. The decreased intensity of the amide bands could be interpreted as the result of alteration of the protein synthesis due to the high affinity of metal compounds towards different amino acid residues of proteins. Further, meso-2, 3 dimercaptosuccinic acid (DMSA) treatment shows the recovery of the protein content in the liver tissues. To confirm that the changes observed are only due to the bio-accumulation of arsenic, the concentration of arsenic in the liver tissues of Labeo rohita was determined by using Inductively Coupled Plasma Optical Emission Spectrometry (ICP-OES). It is observed that the arsenic level in the control tissues is found to be below detectable limit, whereas the arsenic exposed liver shows an accumulation of 66.68 +/- 0.43 MUg/g and DMSA treatment reduces the arsenic content to 17.96 +/- 0.19 MUg/g. In conclusion, this study gives clear evidence that the use of FT-IR spectroscopy is a powerful approach to achieve more insight into the protein alterations caused by arsenic. PMID- 20014410 TI - The role of oxidative stress in Sudan IV-induced DNA damage in human liver derived HepG2 cells. AB - OBJECTIVES: We evaluated the role of oxidative stress in Sudan IV-induced DNA damage, using human liver-derived HepG2 cells. METHODS: The DNA damaging effects of Sudan IV in HepG2 cells were evaluated by alkaline single cell gel electrophoresis assay and micronucleus test (MNT). To clarify the underlying mechanisms, we monitored the intracellular generation of reactive oxygen species (ROS) by 2, 7-dichlorofluorescein diacetate assay and the level of oxidative DNA damage by immunoperoxidase staining for 8-hydroxydeoxyguanosine (8-OHdG). Furthermore, the intracellular glutathione (GSH) level was moderated by pretreatment with buthionine-(S,R)-sulfoximine (BSO), a specific GSH synthesis inhibitor. RESULTS: A significant dose-dependent increment in DNA migration was detected at all tested concentrations (25-100 MUM) of Sudan IV. And in the MNT, a significant increase of the frequency of micronuclei was found at higher tested concentrations (50-100 MUM). The data suggested that Sudan IV caused DNA strand breaks and chromosome breaks. In addition, significantly increased levels of ROS, 8-OHdG formation were observed in HepG2 cells. It was also found that depletion of GSH in HepG2 cells with BSO dramatically increased the susceptibility of HepG2 cells to Sudan IV-induced DNA damage. CONCLUSIONS: Based on these data we believe that Sudan IV exerts toxic effects in HepG2 cells, probably through oxidative DNA damage induced by intracellular ROS and depletion of GSH. PMID- 20014411 TI - Computing chiroptical properties with first-principles theoretical methods: background and illustrative examples. AB - This "tutorial style" review outlines the theoretical foundation for computations of chiroptical properties for optically active molecules. The formalism covers electronic and vibrational CD, optical rotation, and Raman optical activity. The focus is on first-principles methods. A dedicated section highlights the strengths and weaknesses of currently popular time-dependent density functional methods. The article also contains a section with input examples and results for a small molecule (trans-2,3-dimethyloxirane). Several representative applications of computational methods roughly from 2001 to mid-2009 are reviewed in detail. PMID- 20014412 TI - Importance of NADPH supply for improved L-valine formation in Corynebacterium glutamicum. AB - Cofactor recycling is known to be crucial for amino acid synthesis. Hence, cofactor supply was now analyzed for L-valine to identify new targets for an improvement of production. The central carbon metabolism was analyzed by stoichiometric modeling to estimate the influence of cofactors and to quantify the theoretical yield of L-valine on glucose. Three different optimal routes for L-valine biosynthesis were identified by elementary mode (EM) analysis. The modes differed mainly in the manner of NADPH regeneration, substantiating that the cofactor supply may be crucial for efficient L-valine production. Although the isocitrate dehydrogenase as an NADPH source within the tricarboxylic acid cycle only enables an L-valine yield of Y(Val/Glc) = 0.5 mol L-valine/mol glucose (mol Val/mol Glc), the pentose phosphate pathway seems to be the most promising NADPH source. Based on the theoretical calculation of EMs, the gene encoding phosphoglucoisomerase (PGI) was deleted to achieve this EM with a theoretical yield Y(Val/Glc) = 0.86 mol Val/mol Glc during the production phase. The intracellular NADPH concentration was significantly increased in the PGI deficient mutant. L-Valine yield increased from 0.49 +/- 0.13 to 0.67 +/- 0.03 mol Val/mol Glc, and, concomitantly, the formation of by-products such as pyruvate was reduced. PMID- 20014413 TI - Noninvasive prenatal diagnosis: past, present, and future. AB - The presence of fetal cells in the maternal circulation was first noted by Georg Schmorl when he documented the presence of multinucleated syncytial giant cells of placental origin in the lung tissue of women who had died from complications of eclampsia. In the intervening century, advances in cellular and molecular biology further elucidated both the physiology and pathophysiology of communication within the fetomaternal unit. This concept is at the foundation of the rapidly expanding field of noninvasive prenatal diagnosis. However, the clinical utility of this phenomenon had been limited until the presence of cell free fetal DNA circulating in the maternal plasma was reported in 1997 and fetal messenger RNA was demonstrated to circulate in the maternal plasma in 2000. These circulating nucleic acids are found free-floating in the maternal plasma, unencumbered by a surrounding fetal cell. The analysis of these 3 fetal markers (fetal cells, cell-free fetal DNA, and fetal messenger RNA) for diagnostic and screening purposes is now being developed. The scope of noninvasive prenatal diagnosis is not limited to only the diagnosis of fetal genetic traits and aneuploidies. Recently, researchers have focused their investigations on the role of cell-free fetal DNA and fetal messenger RNA in preeclampsia, intrauterine growth restriction, and preterm labor. These biomarkers, the result of inherent placental dysfunction or the byproducts of placental trophoblastic apoptosis, may allow for improvements in the diagnosis and management of high-risk pregnancies. PMID- 20014415 TI - Achieving a safety culture in obstetrics. AB - Preventable maternal and neonatal mortalities still occur, despite the wonders of today's technologically advanced healthcare system. Delivering high-quality, consistent care is the goal of every provider. Yet, obstetrical practice, in which unpredictable events and high-risk situations are the norm, is particularly vulnerable to medical errors. Obstetrics departments should be striving for a climate of patient safety, one that includes a just, reporting, and learning culture. This article discusses the various components of a safety culture as well as some of the advances that are being made in the field to improve the quality of care in obstetrics. PMID- 20014416 TI - A comprehensive approach to the treatment of uterine leiomyomata. AB - Leiomyomas (fibroids) are the most common tumors in women, with a prevalence between 30% and 50%. They affect women primarily during their reproductive years, spontaneously regressing after menopause in most women. They may cause significant symptoms of pain, dysmenorrhea, abnormal uterine bleeding, and infertility. Because leiomyomas are so common, treatment should be reserved for those patients with symptoms. Treatment options have recently expanded beyond hysterectomy. Medical therapies, including gonadotropin-releasing hormone agonists and progesterone modulators, have become more widely used. Less invasive options such as uterine fibroid embolization, magnetic resonance imaging-guided focused ultrasound, and radiofrequency ablation are being used to avoid more invasive surgery. Because of limited and negative information regarding these alternatives to surgery, they are not recommended for women desiring future fertility. If surgery is desired or required, often less invasive approaches via hysteroscopy for intracavitary lesions or robot-assisted laparoscopy for patients with a small number of myomas have become preferred options. Treatment should be tailored to the patient. PMID- 20014417 TI - Bloodless surgery in gynecologic oncology. AB - Bloodless medicine and surgery is an evolving field in the practice of medicine designed to avoid allogeneic transfusions. Although this field has largely developed in response to the growing needs of Jehovah's Witness patients refusing transfusions, all patients may potentially benefit from the avoidance of transfusions. The applications of bloodless techniques and strategies in the field of gynecologic oncology have been limited until recently, in part because of the generally large blood loss associated with gynecologic cancer surgeries. However, as advances in our understanding of bloodless surgery have developed and surgical techniques have been refined, the gynecologic oncology patient can now benefit from the avoidance of allogeneic transfusions. This review outlines bloodless surgery as it applies to gynecologic oncology patients and presents a structured approach to successfully managing these complex patients. PMID- 20014418 TI - Obesity and pregnancy: implications and management strategies for providers. AB - Obesity in pregnancy (pregravid body mass > or =30) has been linked to several adverse pregnancy outcomes, including spontaneous abortion, preeclampsia, gestational diabetes, fetal macrosomia, cesarean delivery, and wound complications post-cesarean section. Intrapartum and postpartum management of obese gravidas requires multidisciplinary consultations between obstetricians, anesthesiologists, nurses, and pediatricians in order to improve the pregnancy outcomes of the mother and neonate. The American College of Obstetricians and Gynecologists currently supports risk-reducing strategies for obese pregnant patients, including limiting weight gain to 15 lb (standardized by the Institute of Medicine). Interventions to reduce gestational weight gain may be important modifiable risk factors for maternal and fetal perinatal complications. Interventions have targeted modifications of diet and exercise with educational methods such as radio broadcasts, pamphlets, and counseling. Interventions have also focused on motivational methods, such as individual and group classes, and have been implemented both before conception and immediately after birth. Effective interventions appear to be individualized in approach, but there is a lack of data to support any specific model. Prospective interventional studies are needed to demonstrate the benefits of weight limitation on pregnancy outcomes. PMID- 20014419 TI - Advances and controversies in assisted reproductive technology. AB - As the incidence and awareness of infertility have increased in the last decades, reproductive endocrinologists and scientists have striven to improve the therapeutic options being offered to patients. One of the most advanced and efficient technologies currently being offered is assisted reproductive technology. Among the various techniques that are comprised by assisted reproductive technology, in vitro fertilization is the most widely studied and used, being responsible for approximately 1% of all live births in the United States. As this technology has evolved, many controversies have arisen, and it is the purpose of this article to review what in the authors' opinion are the most current and controversial aspects of the whole process of in vitro fertilization and its related techniques. PMID- 20014420 TI - Application of agent-based system for bioprocess description and process improvement. AB - Modeling plays an important role in bioprocess development for design and scale up. Predictive models can also be used in biopharmaceutical manufacturing to assist decision-making either to maintain process consistency or to identify optimal operating conditions. To predict the whole bioprocess performance, the strong interactions present in a processing sequence must be adequately modeled. Traditionally, bioprocess modeling considers process units separately, which makes it difficult to capture the interactions between units. In this work, a systematic framework is developed to analyze the bioprocesses based on a whole process understanding and considering the interactions between process operations. An agent-based approach is adopted to provide a flexible infrastructure for the necessary integration of process models. This enables the prediction of overall process behavior, which can then be applied during process development or once manufacturing has commenced, in both cases leading to the capacity for fast evaluation of process improvement options. The multi-agent system comprises a process knowledge base, process models, and a group of functional agents. In this system, agent components co-operate with each other in performing their tasks. These include the description of the whole process behavior, evaluating process operating conditions, monitoring of the operating processes, predicting critical process performance, and providing guidance to decision-making when coping with process deviations. During process development, the system can be used to evaluate the design space for process operation. During manufacture, the system can be applied to identify abnormal process operation events and then to provide suggestions as to how best to cope with the deviations. In all cases, the function of the system is to ensure an efficient manufacturing process. The implementation of the agent-based approach is illustrated via selected application scenarios, which demonstrate how such a framework may enable the better integration of process operations by providing a plant-wide process description to facilitate process improvement. PMID- 20014421 TI - Current surgical management of ulcerative colitis. AB - Inflammatory bowel disease is divided into 2 major disease entities: Crohn's disease and ulcerative colitis. Ulcerative colitis is characterized by contiguous inflammation of the colorectal mucosa, always beginning in and involving the rectum and progressing for variable distances proximally within the colon. In ulcerative colitis, medical therapy, which is not curative, is directed at controlling symptoms and reducing the underlying inflammatory process. However, emergent or elective removal of the colon and rectum does cure the disease and also eliminates the possibility of developing a malignancy. Here we present the current surgical treatment of ulcerative colitis and issues in the management of ulcerative colitis. We discuss indications for surgical treatment, elective and emergent operative management, early and late complications of surgery, and functional results. PMID- 20014423 TI - Recent advances in the surgical management of cervical cancer. AB - Recent advances in the surgical management of early cervical cancer, including abdominal, laparoscopic, vaginal, and robotic approaches to radical hysterectomy as well as fertility-sparing radical trachelectomy, are reviewed. The nerve sparing abdominal radical hysterectomy technique allows for a significant reduction in postoperative bladder morbidity. Radical vaginal hysterectomy with laparoscopic lymph node dissection is a well-recognized technique that offers excellent cure rates without abdominal entry as well as reduced postoperative febrile and gastrointestinal morbidity. Total laparoscopic radical hysterectomy is a minimally invasive alternative to the traditional abdominal radical hysterectomy approach and yields a comparable safety profile with a significant reduction in blood loss and hospital stay. Robotic surgery is becoming more widely accepted in the management of gynecologic cancers, including radical hysterectomy for early cervical cancer. Young women desiring to bear children in the future may be candidates for fertility preservation options, and the radical trachelectomy operation has been described and performed with abdominal, vaginal, laparoscopic, and robotic techniques. There are a number of surgical options for the treatment of women with early cervical cancer. The feasibility and safety of some of these techniques have been well established, whereas for others, the oncological outcome data are only preliminary. The decision to use newer techniques should be directed by patient variables as well as the surgeon's training and competence with laparoscopy, robotics, or vaginal surgery. PMID- 20014422 TI - Generation of an EphA4 conditional allele in mice. AB - Ephrins and Eph receptor tyrosine kinases are cell-surface molecules that serve a multitude of functions in cell-cell communication in development, physiology, and disease. EphA4 is a promiscuous member of the EphA subclass of Eph receptors and can bind to both EphrinAs and EphrinBs. In addition to its well-established roles in guiding the development of neuronal connectivity, EphA4 has been implicated for a role in synaptic plasticity, vascular formation, axon regeneration, and central nervous system repair following injury. However, the study of its role in the adult stage has been hampered by confounding developmental defects in EphA4 germline mutants. Here, we report the generation and molecular characterization of an EphA4 conditional allele along with a novel null allele with a knockin fluorescent reporter gene (mCFP). The conditional allele will be useful in ascertaining postdevelopmental and/or cell type-specific function of EphA4 in physiology, injury, and disease. PMID- 20014425 TI - Genomics of cardiovascular disease. AB - As the leading cause of death worldwide and a major cause of disability, cardiovascular disease remains a central focus of basic research, pharmacological treatment, surgical interventions, and long-term care. Inherited, monogenic syndromes have provided insight into pathophysiological mechanisms across the range of cardiovascular diseases. With the advent of post-Human Genome Project resources and technology, there has been a flood of research aimed at genome-wide predisposition markers, pharmacogenetics, and genomic signatures in complex cardiovascular disorders. Genomic research has both further elucidated the impact of genes previously identified in cardiovascular disease development and progression and discovered genomic regions as yet unknown to be associated with cardiovascular outcomes. The promise of personalized medicine lies in combining this genetic information with other biomarkers to tailor preventive and therapeutic strategies to individual patients for effective management, fewer adverse events, and preventive care. PMID- 20014424 TI - Emerging roles of Kruppel-like factor 6 and Kruppel-like factor 6 splice variant 1 in ovarian cancer progression and treatment. AB - Epithelial ovarian cancer is one of the most lethal gynecologic cancers and the fifth most frequent cause of female cancer deaths in the United States. Despite dramatic treatment successes in other cancers through the use of molecular agents targeted against genetically defined events driving cancer development and progression, very few insights into epithelial ovarian cancer have been translated from the laboratory to the clinic. If advances are to be made in the early diagnosis, prevention, and treatment of this disease, it will be critical to characterize the common and private (personalized) genetic defects underlying the development and spread of epithelial ovarian cancer. The tumor suppressor Kruppel-like factor 6 and its alternatively spliced, oncogenic isoform, Kruppel like factor 6 splice variant 1, are members of the Kruppel-like zinc finger transcription factor family of proteins, which have diverse roles in cellular differentiation, development, proliferation, growth-related signal transduction, and apoptosis. Inactivation of Kruppel-like factor 6 and overexpression of Kruppel-like factor 6 splice variant 1 have been associated with the progression of a number of human cancers and even with patient survival. This article summarizes our recent findings demonstrating that a majority of epithelial ovarian cancer tumors have Kruppel-like factor 6 allelic loss and decreased expression coupled with increased expression of Kruppel-like factor 6 splice variant 1. The targeted reduction of Kruppel-like factor 6 in ovarian cancer cell lines results in marked increases in cell proliferation, invasion, tumor growth, angiogenesis, and intraperitoneal dissemination in vivo. In contrast, the inhibition of Kruppel-like factor 6 splice variant 1 decreases cellular proliferation, invasion, angiogenesis, and tumorigenicity; this provides the rationale for its potential therapeutic application. These results and our recent demonstration that the inhibition of Kruppel-like factor 6 splice variant 1 can dramatically prolong survival in a preclinical mouse model of ovarian cancer are reviewed and discussed. PMID- 20014426 TI - Breast magnetic resonance imaging: an overview for nonradiologists. AB - Magnetic resonance imaging is a major component of breast imaging. Many studies have shown that magnetic resonance imaging is the most sensitive imaging method for detecting invasive breast cancer in comparison with mammography, ultrasound, and clinical breast examinations. Evidence-based clinical indications for breast magnetic resonance imaging include screening patients at high risk for breast cancer, including those with breast/ovarian cancer genes (BRCA1 and BRCA2), those who are untested first-degree relatives of carriers of these genes, those whose lifetime risk of developing breast cancer is 20% to 25% or greater, those who had chest radiation when they were 10 to 30 years old, and those who have or are first-degree relatives of people with Li-Fraumeni syndrome, Cowden syndrome, or Bannayan-Riley-Ruvalcaba syndrome. Breast magnetic resonance imaging is performed in conjunction with mammography and does not replace mammography. Outside of the screening population, utilization of breast magnetic resonance imaging for newly diagnosed breast cancer patients and its use as a problem-solving technique for equivocal mammographic or clinical findings remain controversial. An understanding of the current evidence facilitates appropriate utilization of this important medical resource. This article discusses indications for ordering breast magnetic resonance imaging and how to read the breast magnetic resonance imaging report and understand the lexicon used. PMID- 20014427 TI - Chemotherapy for gynecologic cancers. AB - This review highlights significant recent developments and trends in chemotherapy for major gynecologic malignancies, i.e., ovarian cancer, endometrial cancer, uterine sarcomas, gestational trophoblastic neoplasia, and cervical cancer. In ovarian cancer, chemotherapeutic options for early, advanced and recurrent disease are in the adjuvant setting as well as in the neoadjuvant setting are explored. For uterine cancer, adjuvant chemotherapy is employed for high risk epithelial subtypes with early disease, such as uterine papillary serous carcinomas, uterine carcinosarcomas and leiomyosarcomas, advanced stage cases, as well as recurrent disease. The review then proceeds to further discuss the appropriate treatment based on the International Federation of Gynecology and Obstetrics prognostic scoring system for gestational trophoblastic neoplasia. Finally, chemotherapy is utilized in cervical cancer as neo-adjuvant therapy prior to surgery or radiation, as a sensitizer concomitantly with radiation therapy or for the treatment of advanced and recurrent disease. PMID- 20014428 TI - Species-specific interaction of HIV protease inhibitors with accumulation of cholyl-glycylamido-fluorescein (CGamF) in sandwich-cultured hepatocytes. AB - Using sandwich-cultured hepatocytes from rat, dog, pig, and human, we investigated the species-specificity of interaction of HIV protease inhibitors (PI) with in vitro hepatic accumulation of the bile salt analogue cholyl glycylamido-fluorescein (CGamF). Extracellular sodium depletion or coincubation with the OATP/Oatp inhibitors rifampicin and digoxin revealed that about 35% of active CGamF accumulation was mediated by Ntcp/NTCP in rat and human hepatocytes, while the contribution of this sodium-dependent transporter reached 50-60% in dog and pig hepatocytes. One or more sodium-independent transporters, likely belonging to the Oatp/OATP family, constitute a major transport mechanism for CGamF accumulation. Various HIV PI (0.5, 5, 25 microM) exhibited pronounced species differences in their interaction with active CGamF accumulation (1 microM), although some similarity was observed between the dog and human interaction profiles when HIV PI were tested at 0.5 microM. Atazanavir, indinavir, and darunavir were the most potent inhibitors of CGamF accumulation in human hepatocytes. Potent inhibition of CGamF accumulation by ritonavir in rat hepatocytes contrasted with a weak effect in human hepatocytes. Thorough characterization of in vitro disposition of probe substrates in preclinical species compared to human hepatocytes will ultimately support a better insight in species-specific mechanisms underlying drug interactions and drug-mediated toxicity. PMID- 20014429 TI - A mathematical relationship for hydromorphone loading into liposomes with trans membrane ammonium sulfate gradients. AB - We have studied the loading of the opioid hydromorphone into liposomes using ammonium sulfate gradients. Unlike other drugs loaded with this technique, hydromorphone is freely soluble as the sulfate salt, and, consequently, does not precipitate in the liposomes after loading. We have derived a mathematical relationship that can predict the extent of loading based on the ammonium ion content of the liposomes and the amount of drug added for loading. We have adapted and used the Berthelot indophenol assay to measure the amount of ammonium ions in the liposomes. Plots of the inverse of the fraction of hydromorphone loaded versus the amount of hydromorphone added are linear, and the slope should be the inverse of the amount of ammonium ions present in the liposomes. The inverse of the slopes obtained closely correspond to the amount of ammonium ions in the liposomes measured with the Berthelot indophenol assay. We also show that loading can be less than optimal under conditions where osmotically driven loss of ammonium ions or leakage of drug after loading may occur. PMID- 20014430 TI - Enantioselective oxidation of 2-hydroxy carboxylic acids by glycolate oxidase and catalase coexpressed in methylotrophic Pichia pastoris. AB - Glycolate oxidase (GO; (S)-2-hydroxyacid oxidase, EC 1.1.3.15) is a flavin mononucleotide (FMN)-dependent enzyme, which catalyzes the oxidation of 2-hydroxy carboxylic acids to the corresponding 2-keto acids. Catalase has been used as cocatalyst to decompose hydrogen peroxide produced in the reaction, thus limiting peroxide-based side reactions and GO deactivation. GO from spinach and catalase T from Saccharomyces cerevisiae previously coexpressed in Pichia pastoris strain NRRL Y-21001, was permeabilized and used for the oxidation of 3-phenyllactic acid, 3-indolelactic acid, 3-chlorolactic acid, 2-hydroxybutanoic acid, and 2 hydroxydecanoic acid to demonstrate high degree of selectivity to the (S) enantiomers, leaving (R)-isomers intact. The rates of oxidation ranged from 1.3 to 120.0%, relative to the oxidation of lactic acid to pyruvic acid. The best substrates were 3-chlorolactic acid (110%) and 2-hydroxybutanoic acid (120%). Oxidation was carried out with (R)-, (S)-, and (RS)-3-phenyllactic acid, (RS) lactic acid, and (RS)-2-hydroxybutanoic acid in 500 mL scale to characterize the products and stoichiometry of the reaction. All (RS)- and (S)-2-hydroxy acids produced 2-keto acids at close to the theoretical yield in 1-9 h. (R)-3 Phenyllactic acid was not oxidized over a period of 9 h. Addition of exogenous FMN and catalase were not required for this oxidation using double recombinant Pichia pastoris whole cells. As GO is absolutely specific to (S)-enantiomers, it can be used for resolution of racemic 2-hydroxy acids to (R)-2-hydroxy acids as well as for production of 2-keto acids. This is the first report on the selectivity of a broad range of 2-hydroxy acids by GO. PMID- 20014431 TI - Impact of membrane properties on uptake and transcytosis of colloidal nanocarriers across an epithelial cell barrier model. AB - The aim of this study was to investigate the effect of liposomal membrane properties on cellular uptake and transcytosis across a tight Madin-Darby canine kidney (MDCK) cell barrier in vitro. More than 25 small vesicles were prepared by lipid film hydration/extrusion to generate small unilamellar vesicles. The fluorescence marker calcein was encapsulated to mimic hydrophilic drug transport. Marker uptake by MDCK cells seems to be mediated by different mechanisms for the liposomes used. It was mainly depending on membrane fluidity and vesicle charge. Liposomes L2 with a positive charge (325 +/- 3 pmol/well) and vesicles L3 containing the helper lipid dioleylphosphatidylethanolamine (DOPE) in their membrane (216 +/- 42 pmol/well) were taken up to the most. Selected liposomes were tested for their transcytotic transport across a MDCK monolayer. Liposomes L4 containing equimolar DOPE and octadecyl-1,1-dimethylpiperidin-1-ium-4-yl phosphate (OPP) were the most efficient vesicles for transcellular transport resulting in 808 +/- 30 pmol calcein/cm(2) in the basal medium (28.1% of total liposomal marker added). Transcytosis was positively correlated with membrane fluidity in the outer part of the bilayer, as electron paramagnetic resonance measurements revealed. We expect that an increase in membrane fluidity of vesicles should also improve the restricted transport of hydrophilic drugs across the blood-brain barrier. PMID- 20014432 TI - Functionalization of cellulose acetate fibers with engineered cutinases. AB - In the present work, we describe for the first time the specific role of cutinase on surface modification of cellulose acetate fibers. Cutinase exhibits acetyl esterase activity on diacetate and triacetate of 0.010 U and 0.007 U, respectively. An increase on the hydroxyl groups at the fiber surface of 25% for diacetate and 317% for triacetate, after a 24 h treatment, is estimated by an indirect assay. Aiming at further improvement of cutinase affinity toward cellulose acetate, chimeric cutinases are genetically engineered by fusing the 3' end coding sequence with a bacterial or a fungal carbohydrate-binding module and varying the linker DNA sequence. A comparative analysis of these genetic constructions is presented showing that, the superficial regeneration of cellulose hydrophilicity and reactivity on highly substituted cellulose acetates is achieved by chimeric cutinases. PMID- 20014434 TI - Fusion partners can increase the expression of recombinant interleukins via transient transfection in 2936E cells. AB - The expression levels of five secreted target interleukins (IL-11, 15, 17B, 32, and IL23 p19 subunit) were tested with three different fusion partners in 2936E cells. When fused to the N-terminus, human serum albumin (HSA) was found to enhance the expression of both IL-17B and IL-15, cytokines which did not express at measurable levels on their own. Although the crystallizable fragment of an antibody (Fc) was also an effective fusion partner for IL-17B, Fc did not increase expression of IL-15. Fc was superior to HSA for the expression of the p19 subunit of IL-23, but no partner led to measurable levels of IL-32gamma secretion. Glutathione S-transferase (GST) did not enhance the expression of any target and suppressed the production of IL-11, a cytokine which expressed robustly both on its own and when fused to HSA or Fc. Cleavage of the fusion partner was not always possible. The use of HSA or Fc as N-terminal fusions can be an effective technique to express difficult proteins, especially for applications in which the fusion partner need not be removed. PMID- 20014435 TI - Characterization of site-directed mutants of residues R58, R59, D116, W340 and R372 in the active site of E. coli cystathionine beta-lyase. AB - Cystathionine beta-lyase (CBL) catalyzes the hydrolysis of L-cystathionine (L Cth) to produce L-homocysteine, pyruvate, and ammonia. A series of active-site mutants of Escherichia coli CBL (eCBL) was constructed to investigate the roles of residues R58, R59, D116, W340, and R372 in catalysis and inhibition by aminoethoxyvinylglycine (AVG). The effects of these mutations on the k(cat)/K(m) (L-Cth) for the beta-elimination reaction range from a reduction of only 3-fold for D116A and D116N to 6 orders of magnitude for the R372L and R372A mutants. The order of importance of these residues for the hydrolysis of L-Cth is: R372 >> R58 > W340 approximately R59 > D116. Comparison of the kinetic parameters for L-Cth hydrolysis with those for inhibition of eCBL by AVG demonstrates that residue R58 tethers the distal carboxylate group of the substrate and confirms that residues W340 and R372 interact with the alpha-carboxylate moiety. The increase in the pK(a) of the acidic limb and decrease in the pK(a) of the basic limb of the k(cat)/K(m) (L-Cth) versus pH profiles of the R58K and R58A mutants, respectively, support a role for this residue in modulating the pK(a) of an active-site residue. PMID- 20014436 TI - Association of high-dose bifonazole administration during early pregnancy and severe limb reduction defects in the newborn. AB - BACKGROUND: Neonatal limb reduction defects may be caused by exposure to an external agent. The azole derivatives are used in the treatment of systemic and dermal mycoses. Their relative teratogenic risk is still controversial. CASES: We describe two newborns with severe limb defects who were exposed to high doses of oral (an unacceptable route) and/or intravaginal bifonazole during the entire first trimester of pregnancy. CONCLUSION: Although only two cases are insufficient to establish a relationship, our data suggest that maternal intake of bifonazole in early pregnancy poses a risk of morphogenic malformations. The literature suggests several possible mechanisms. PMID- 20014437 TI - Influence of substrate stiffness on the phenotype of heart cells. AB - Adult cardiomyocytes (CM) retain little capacity to regenerate, which motivates efforts to engineer heart tissues that can emulate the functional and mechanical properties of native myocardium. Although the effects of matrix stiffness on individual CM have been explored, less attention was devoted to studies at the monolayer and the tissue level. The purpose of this study was to characterize the influence of substrate mechanical stiffness on the heart cell phenotype and functional properties. Neonatal rat heart cells were seeded onto collagen-coated polyacrylamide (PA) substrates with Young's moduli of 3, 22, 50, and 144 kPa. Collagen-coated glass coverslips without PA represented surfaces with effectively "infinite" stiffness. The local elastic modulus of native neonatal rat heart tissue was measured to range from 4.0 to 11.4 kPa (mean value of 6.8 kPa) and for native adult rat heart tissue from 11.9 to 46.2 kPa (mean value of 25.6 kPa), motivating our choice of the above PA gel stiffness. Overall, by 120 h of cultivation, the lowest stiffness PA substrates (3 kPa) exhibited the lowest excitation threshold (ET; 3.5 +/- 0.3 V/cm), increased troponin I staining (52% positively stained area) but reduced cell density, force of contraction (0.18 +/- 0.1 mN/mm(2)), and cell elongation (aspect ratio = 1.3-1.4). Higher stiffness (144 kPa) PA substrates exhibited reduced troponin I staining (30% positively stained area), increased fibroblast density (70% positively stained area), and poor electrical excitability. Intermediate stiffness PA substrates of stiffness comparable to the native adult rat myocardium (22-50 kPa) were found to be optimal for heart cell morphology and function, with superior elongation (aspect ratio > 4.3), reasonable ET (ranging from 3.95 +/- 0.8 to 4.4 +/- 0.7 V/cm), high contractile force development (ranging from 0.52 +/- 0.2 to 1.60 +/- 0.6 mN/mm(2)), and well-developed striations, all consistent with a differentiated phenotype. PMID- 20014438 TI - Hyperosmotic stress induces autophagy and apoptosis in recombinant Chinese hamster ovary cell culture. AB - During recombinant Chinese hamster ovary (rCHO) cell culture, various events, such as feeding with concentrated nutrient solutions or the addition of base to maintain an optimal pH, increase the osmolality of the medium. To determine the effect of hyperosmotic stress on two types of programmed cell death (PCD), apoptosis and autophagy, of rCHO cells, two rCHO cell lines, producing antibody and erythropoietin, were subjected to hyperosmotic stress resulting from NaCl addition (310-610 mOsm/kg). For both rCHO cell lines, hyperosmolality up to 610 mOsm/kg increased cleaved forms of PARP, caspase-3, caspase-7, and fragmentation of chromosomal DNA, confirming the previous observation that apoptosis was induced by hyperosmotic stress. Concurrently, hyperosmolality increased the level of accumulation of LC3-II, a widely used autophagic marker, which was determined by Western blot analysis and confocal microscopy. When glucose and glutamine concentrations were measured during the cultures, glucose and glutamine concentrations in the culture medium at various osmolalities (310-610 mOsm/kg) showed no significant differences. This result suggests that induction of PCD by hyperosmotic stress occurred independently of nutrient depletion. Taken together, autophagy as well as apoptosis was observed in rCHO cells subjected to hyperosmolality. PMID- 20014439 TI - Semi-permeable membrane retention of synovial fluid lubricants hyaluronan and proteoglycan 4 for a biomimetic bioreactor. AB - Synovial fluid (SF) contains lubricant macromolecules, hyaluronan (HA), and proteoglycan 4 (PRG4). The synovium not only contributes lubricants to SF through secretion by synoviocyte lining cells, but also concentrates lubricants in SF due to its semi-permeable nature. A membrane that recapitulates these synovium functions may be useful in a bioreactor system for generating a bioengineered fluid (BF) similar to native SF. The objectives were to analyze expanded polytetrafluoroethylene membranes with pore sizes of 50 nm, 90 nm, 170 nm, and 3 microm in terms of (1) HA and PRG4 secretion rates by adherent synoviocytes, and (2) the extent of HA and PRG4 retention with or without synoviocytes adherent on the membrane. Experiment 1: Synoviocytes were cultured on tissue culture (TC) plastic or membranes +/- IL-1beta + TGF-beta1 + TNF-alpha, a cytokine combination that stimulates lubricant synthesis. HA and PRG4 secretion rates were assessed by analysis of medium. Experiment 2: Bioreactors were fabricated to provide a BF compartment enclosed by membranes +/- adherent synoviocytes, and an external compartment of nutrient fluid (NF). A solution with HA (1 mg/mL, MW ranging from 30 to 4,000 kDa) or PRG4 (50 microg/mL) was added to the BF compartment, and HA and PRG4 loss into the NF compartment after 2, 8, and 24 h was determined. Lubricant loss kinetics were analyzed to estimate membrane permeability. Experiment 1: Cytokine-regulated HA and PRG4 secretion rates on membranes were comparable to those on TC plastic. Experiment 2: Transport of HA and PRG4 across membranes was lowest with 50 nm membranes and highest with 3 microm membranes, and transport of high MW HA was decreased by adherent synoviocytes (for 50 and 90 nm membranes). The permeability to HA mixtures for 50 nm membranes was approximately 20 x 10(-8) cm/s (- cells) and approximately 5 x 10(-8) cm/s (+ cells), for 90 nm membranes was approximately 35 x 10(-8) cm/s (- cells) and approximately 19 x 10(-8) cm/s (+ cells), for 170 nm membranes was approximately 74 x 10(-8) cm/s (+/- cells), and for 3 microm membranes was approximately 139 x 10(-8) cm/s (+/- cells). The permeability of 450 kDa HA was approximately 40x lower than that of 30 kDa HA for 50 nm membranes, but only approximately 2.5x lower for 3 microm membranes. The permeability of 4,000 kDa HA was approximately 250x lower than that of 30 kDa HA for 50 nm membranes, but only approximately 4x lower for 3 microm membranes. The permeability for PRG4 was approximately 4 x 10( 8) cm/s for 50 nm membranes, approximately 48 x 10(-8) cm/s for 90 nm membranes, approximately 144 x 10(-8) cm/s for 170 nm membranes, and approximately 336 x 10( 8) cm/s for 3 microm membranes. The associated loss across membranes after 24 h ranged from 3% to 92% for HA, and from 3% to 93% for PRG4. These results suggest that semi-permeable membranes may be used in a bioreactor system to modulate lubricant retention in a bioengineered SF, and that synoviocytes adherent on the membranes may serve as both a lubricant source and a barrier for lubricant transport. PMID- 20014440 TI - Aggregation of a multidomain protein: a coagulation mechanism governs aggregation of a model IgG1 antibody under weak thermal stress. AB - Using an IgG1 antibody as a model system, we have studied the mechanisms by which multidomain proteins aggregate at physiological pH when incubated at temperatures just below their lowest thermal transition. In this temperature interval, only minor changes to the protein conformation are observed. Light scattering consistently showed two coupled phases: an initial fast phase followed by several hours of exponential growth of the scattered intensity. This is the exact opposite of the lag-time behavior typically observed in protein fibrillation. Dynamic light scattering showed the rapid formation of an aggregate species with a hydrodynamic radius of about 25 nm, which then increased in size throughout the experiment. Theoretical analysis of our light scattering data showed that the aggregate number density goes through a maximum in time providing compelling evidence for a coagulation mechanism in which aggregates fuse together. Both the analysis as well as size-exclusion chromatography of incubated samples showed the actual increase in aggregate mass to be linear and reach saturation long before all molecules had been converted to aggregates. The CH2 domain is the only domain partly unfolded in the temperature interval studied, suggesting a pivotal role of this least stable domain in the aggregation process. Our results show that for multidomain proteins at temperatures below their thermal denaturation, transient unfolding of a single domain can prime the molecule for aggregation, and that the formation of large aggregates is driven by coagulation. PMID- 20014441 TI - Sensitive multiplex RNA quantification using capillary electrophoresis-based single-strand conformation polymorphism. AB - Quantification of RNA provides information crucial for various biological studies, including analysis of mRNA expression and that of microRNAs. Reverse transcription (RT) coupled with real-time polymerase chain reaction (PCR) is known to be the most accurate method for quantifying nucleic acids, and thus represents the state-of-the-art for RNA quantification. However, the use of real time PCR for RNA quantification is limited to a single target per analytical run because of reductions in quantification power and limitations of fluorescence dyes associated with multiplex applications. Here, we report a novel multiplex RNA quantification method that uses capillary electrophoresis single-strand conformation polymorphism (CE-SSCP) coupled with modified RT and asymmetric PCR. The reverse transcripts of seven in vitro transcribed RNAs were modified with common sequence tags and amplified by asymmetric PCR using primers specific to the common tags. The resulting amplicons were separated and quantified by CE SSCP. A series of experiments using different amounts of RNA demonstrated that the assay had a limit of detection of 2 amol and a dynamic range of approximately 10(5). These results clearly indicate the potential of this method to provide robust and precise multiplex RNA quantification. PMID- 20014442 TI - Hypoxia enhances proliferation of mouse embryonic stem cell-derived neural stem cells. AB - Neural stem (NS) cells can provide a source of material with potential applications for neural drug testing, developmental studies, or novel treatments for neurodegenerative diseases. Herein, the ex vivo expansion of a model system of mouse embryonic stem (mES) cell-derived NS cells was characterized and optimized, cells being cultivated under adherent conditions. Culture was first optimized in terms of initial cell plating density and oxygen concentration, known to strongly influence brain-derived NS cells. To this end, the growth of cells cultured under hypoxic (2%, 5%, and 10% O(2)) and normoxic (20% O(2)) conditions was compared. The results showed that 2-5% oxygen, without affecting multipotency, led to fold increase values in total cell number about twice higher than observed under 20% oxygen (20-fold vs. 10-fold, respectively) this effect being more pronounced when cells were plated at low density. With an optimal cell density of 10(4) cells/cm(2), the maximum growth rates were 1.9 day(-1) under hypoxia versus 1.7 day(-1) under normoxia. Cell division kinetics analysis by flow cytometry based on PKH67 tracking showed that when cultured in hypoxia, cells are at least one divisional generation ahead compared to normoxia. In terms of cell cycle, a larger population in a quiescent G(0) phase was observed in normoxic conditions. The optimization of NS cell culture performed here represents an important step toward the generation of a large number of neural cells from a reduced initial population, envisaging the potential application of these cells in multiple settings. PMID- 20014443 TI - Elucidating residue roles in engineered variants of AraC regulatory protein. AB - The AraC regulatory protein was previously engineered to control gene expression specifically in response to D-arabinose and not the native effector L-arabinose (Tang et al., J Am Chem Soc 2008;130:5267-5271). Mutations were targeted in the ligand-binding pocket and on the AraC N-terminal arm, which plays an important role in maintaining repressing or activating conformations in the absence or presence of effector, respectively. In this study, we analyze the contributions of individual mutations toward the overall mutant functions in an attempt to streamline future AraC design efforts. For a variety of point mutants, we quantify the induced expression response to D-arabinose (level of leaky expression, induction fold, half-maximal dose response, and effector specificity) and the binding affinity of the purified ligand-binding domain for D-arabinose. We find that mutations introduced in the N-terminal arm (design Position 8) strengthen the induction response, most likely by weakening interactions with the DNA-binding domain, but are not involved in ligand binding. Meanwhile, binding pocket mutations occurring further away from the arm (Positions 80 and 82) primarily contribute to maintaining repression in the absence of effector and do not show response to D-arabinose without the accompanying mutations. Combinations of mutations cooperatively couple molecular recognition to transcriptional activation, demonstrating the complexity of the AraC regulatory switch and the power of combinatorial protein design to alter effector specificity while maintaining regulatory function. PMID- 20014444 TI - Mechanism of glyceraldehyde-3-phosphate dehydrogenase inactivation by tyrosine nitration. AB - Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) is a multifaceted protein that is involved in numerous processes including glycolysis, translational silencing, transcriptional regulation of specific genes, and acting as a nitric oxide sensor. The precise mechanism on how GAPDH is targeted to these different roles is unclear but believed to involve specific posttranslational modification to the protein. Numerous studies have demonstrated that GAPDH is a target for tyrosine nitration. However, the site of modification and the molecular consequence have not been defined. Rabbit GAPDH with a reversibly protected catalytic cysteine was nitrated in vitro with tetranitromethane, resulting in complete loss of GAPDH catalytic activity. Nitration was estimated as 0.32 mol of nitrotyrosine residue per mole of GAPDH. Mass spectrometry analysis of nitrated GAPDH indicated that Tyr311 and Tyr317 were the sole sites of nitration. The X-ray crystal structure revealed that the distances between Tyr311 and Tyr317 and the cofactor nicotinamide adenine dinucleotide (NAD(+)) were less than 7.2 and 3.7 A, respectively, implying that nitration of these two residues may affect NAD(+) binding. This possibility was assessed using an NAD(+) binding assay, which showed that nitrated GAPDH was incapable of binding NAD(+). Thus, these results strongly suggest that Tyr311 and Tyr317 nitration prohibits NAD(+) binding, leading to the loss of catalytic activity. PMID- 20014445 TI - Germline humanization of a murine Abeta antibody and crystal structure of the humanized recombinant Fab fragment. AB - Alzheimer's disease is the most common form of dementia, affecting 26 million people worldwide. The Abeta peptide (39-43 amino acids) derived from the proteolytic cleavage of the amyloid precursor protein is one of the main constituents of amyloid plaques associated with disease pathogenesis and therefore a validated target for therapy. Recently, we characterized antibody fragments (Fab and scFvs) derived from the murine monoclonal antibody WO-2, which bind the immunodominant epitope ((3)EFRH(6)) in the Abeta peptide at the N terminus. In vitro, these fragments are able to inhibit fibril formation, disaggregate preformed amyloid fibrils, and protect neuroblastoma cells against oligomer-mediated toxicity. In this study, we describe the humanization of WO-2 using complementary determining region loop grafting onto the human germline gene and the determination of the three-dimensional structure by X-ray crystallography. This humanized version retains a high affinity for the Abeta peptide and therefore is a potential candidate for passive immunotherapy of Alzheimer's disease. PMID- 20014446 TI - Microsurgical free flap in head and neck reconstruction. AB - Microsurgical free flaps are today considered state of the art in head and neck reconstruction after composite tumor resections. Free flaps provide superior functional and aesthetic restoration with less donor-site morbidity. This article details our approach to this challenging and complex procedure. Free tissue transfer can be viewed as consisting of 4 essential stages: (1) defect assessment, (2) preparation of recipient vessels, (3) flap selection and harvest, and (4) flap inset and microsurgical anastomoses. The essential details of each step are highlighted. Meticulous attention to each step is important because each plays a crucial role in the overall success of the procedure. Workhorse flaps in our practice are the anterolateral thigh, radial forearm, fibula, and jejunum flaps. Unique issues related to postoperative care and monitoring of head and neck free flaps are discussed. The management of complications, in particular those threatening flap survival, are reviewed in detail. PMID- 20014447 TI - Integrative molecular characterization of head and neck cancer cell model genomes. AB - BACKGROUND: Cell lines are invaluable model systems for the investigation of cancer. Knowledge of the molecular alterations that exist within cell models is required to define the mechanisms governing cellular phenotypes. METHODS: Five tongue squamous cell carcinomas cell lines and 1 submaxillary salivary gland epidermoid carcinoma cell line were analyzed for copy number and mRNA expression by tiling-path DNA microarrays and Agilent Whole Human Genome Oligoarrays, respectively. RESULTS: Integrative analysis of genetic and expression alterations revealed the molecular landscape of each cell line. Molecular results for individual cell lines and across all samples have been summarized and made available for easy reference. CONCLUSION: Our integrative genomic analyses have defined the DNA and RNA alterations for each individual line. These data will be useful to anyone modeling oral cancer behavior, providing a molecular context that will be useful for deciphering cell phenotypes. PMID- 20014448 TI - The effect of the deliberation process and jurors' prior legal knowledge on the sentence: the role of psychological expertise and crime scene photo. AB - An experiment with simulated juries (N = 198) tested the impact of the deliberation process and two extra legal variables on the determination of sentence. Participants were either social science students without prior instruction in criminal law (prior legal knowledge: low-level group) or future professional magistrates completing their final year of training (high-level group). We manipulated the presence versus absence of (i) a non-diagnostic observation of the defendant by a psychology expert and (ii) a realistic crime scene photograph. After controlling for participants' gender and age, our results show that the high-level group was both less sensitive to the manipulated variables and more severe in their sentence than low-level jurors. We observed a post-deliberation increase in pre-deliberation bias such that the non-diagnostic psychological expertise had a stronger post-deliberation impact on the sentence. Finally, an unexpected effect showed that aggressive responses during the psychological observation tended to operate as exculpatory rather than accusatory evidence. Our results are discussed on the basis of previous research and proposals for future research are made. PMID- 20014451 TI - Real-time observation of Wnt beta-catenin signaling in the chick embryo. AB - A critical mediator of cell-cell signaling events during embryogenesis is the highly conserved Wnt family of secreted proteins. Reporter constructs containing multimerized TCF DNA binding sites have been used to detect Wnt beta-catenin dependent activity during animal development. In this report, we have constructed and compared several TCF green fluorescent protein (GFP) reporter constructs. They contained 3, 8, or 12 TCF binding sites upstream of a minimal promoter driving native or destabilized enhanced GFP (EGFP). We have used the electroporation of somites in the chick embryo as a paradigm to test them in vivo. We have verified that they all respond to Wnt signaling in vivo. We have then assessed their efficiency at reflecting the activity of the Wnt pathway. Using destabilized EGFP reporter constructs, we show that somite cells dynamically regulate Wnt/beta-catenin-dependent signaling, a finding that was confirmed by performing time-lapse video confocal observation of electroporated embryos. PMID- 20014452 TI - Role of transcatheter arterial embolization for massive bleeding from gastroduodenal ulcers. AB - Intractable bleeding from gastric and duodenal ulcers is associated with significant morbidity and mortality. Aggressive treatment with early endoscopic hemostasis is essential for a favourable outcome. In as many as 12%-17% of patients, endoscopy is either not available or unsuccessful. Endovascular therapy with selective catheterization of the culprit vessel and injection of embolic material has emerged as an alternative to emergent operative intervention in high risk patients. There has not been a systematic literature review to assess the role for embolotherapy in the treatment of acute upper gastrointestinal bleeding from gastroduodenal ulcers after failed endoscopic hemostasis. Here, we present an overview of indications, techniques, and clinical outcomes after endovascular embolization of acute peptic-ulcer bleeding. Topics of particular relevance to technical and clinical success are also discussed. Our review shows that transcatheter arterial embolization is a safe alternative to surgery for massive gastroduodenal bleeding that is refractory to endoscopic treatment, can be performed with high technical and clinical success rates, and should be considered the salvage treatment of choice in patients at high surgical risk. PMID- 20014453 TI - Invasive front of colorectal cancer: dynamic interface of pro-/anti-tumor factors. AB - Tumor-host interaction at the invasive front of colorectal cancer represents a critical interface encompassing a dynamic process of de-differentiation of colorectal carcinoma cells known as epithelial mesenchymal transition (EMT). EMT can be identified histologically by the presence of "tumor budding", a feature which can be highly specific for tumors showing an infiltrating tumor growth pattern. Importantly, tumor budding and tumor border configuration have generated considerable interest as additional prognostic factors and are also recognized as such by the International Union Against Cancer. Evidence seems to suggest that the presence of tumor budding or an infiltrating growth pattern is inversely correlated with the presence of immune and inflammatory responses at the invasive tumor front. In fact, several tumor-associated antigens such as CD3, CD4, CD8, CD20, Granzyme B, FOXP3 and other immunological or inflammatory cell types have been identified as potentially prognostic in patients with this disease. Evidence seems to suggest that the balance between pro-tumor (including budding and infiltrating growth pattern) and anti-tumor (immune response or certain inflammatory cell types) factors at the invasive front of colorectal cancer may be decisive in determining tumor progression and the clinical outcome of patients with colorectal cancer. On one hand, the infiltrating tumor border configuration and tumor budding promote progression and dissemination of tumor cells by penetrating the vascular and lymphatic vessels. On the other, the host attempts to fend off this attack by mounting an immune response to protect vascular and lymphatic channels from invasion by tumor buds. Whereas standard pathology reporting of breast and prostate cancer involves additional prognostic features, such as the BRE and Gleason scores, the ratio of pro- and anti-tumor factors could be a promising approach for the future development of a prognostic score for patients with colorectal cancer which could complement tumor node metastasis staging to improve the clinical management of patients with this disease. PMID- 20014454 TI - Colorectal cancer screening in Europe. AB - Colorectal cancer (CRC) is the second most frequent malignant disease in Europe. Every year, 412 000 people are diagnosed with this condition, and 207 000 patients die of it. In 2003, recommendations for screening programs were issued by the Council of the European Union (EU), and these currently serve as the basis for the preparation of European guidelines for CRC screening. The manner in which CRC screening is carried out varies significantly from country to country within the EU, both in terms of organization and the screening test chosen. A screening program of one sort or another has been implemented in 19 of 27 EU countries. The most frequently applied method is testing stool for occult bleeding (fecal occult blood test, FOBT). In recent years, a screening colonoscopy has been introduced, either as the only method (Poland) or the method of choice (Germany, Czech Republic). PMID- 20014455 TI - Progress in researches about focal adhesion kinase in gastrointestinal tract. AB - Focal adhesion kinase (FAK) is a 125-kDa non-receptor protein tyrosine. Growth factors or the clustering of integrins facilitate the rapid phosphorylation of FAK at Tyr-397 and this in turn recruits Src-family protein tyrosine kinases, resulting in the phosphorylation of Tyr-576 and Tyr-577 in the FAK activation loop and full catalytic FAK activation. FAK plays a critical role in the biological processes of normal and cancer cells including the gastrointestinal tract. FAK also plays an important role in the restitution, cell survival and apoptosis and carcinogenesis of the gastrointestinal tract. FAK is over-expressed in cancer cells and its over-expression and elevated activities are associated with motility and invasion of cancer cells. FAK has been proposed as a potential target in cancer therapy. Small molecule inhibitors effectively inhibit the kinase activity of FAK and show a potent inhibitory effect for the proliferation and migration of tumor cells, indicating a high potential for application in cancer therapy. PMID- 20014458 TI - Ketoprofen, peginterferon 2a and ribavirin for genotype 1 chronic hepatitis C: a phase II study. AB - AIM: To evaluate the safety of adding ketoprofen to pegylated-interferon (PEG IFN) with or without ribavirin and the effect on viral kinetics, STAT1 activity and expression of 2'-5'-oligoadenylate synthetase (2'-5'OAS) in genotype 1 chronic hepatitis C in a phase II study. METHODS: Forty-five patients were studied: fifteen were randomized to PEG-IFN plus ribavirin (PR), 16 to PEG-IFN plus ketoprofen and 14 to PR and ketoprofen. The molecular study of IFN-dependent signal transduction was conducted in 9 patients from each group. RESULTS: The combination of ketoprofen and PEG-IFN with or without ribavirin was safe and well tolerated. An early activation of STAT1 was observed in ketoprofen-treated patients, but this activation was less sustained over time. Conversely, ketoprofen plus PEG-IFN and ribavirin induced an early and sustained increase of 2'-5'OAS transcription starting 24 h after the first dose until the 36th wk. These data are consistent with the clinical results, showing a better sustained virological response and a lower relapse rate in patients receiving ketoprofen plus PEG-IFN and ribavirin. CONCLUSION: The addition of ketoprofen to the standard therapy of chronic hepatitis C should be explored in larger randomized clinical studies. PMID- 20014456 TI - TRAIL-induced apoptosis of hepatocellular carcinoma cells is augmented by targeted therapies. AB - AIM: To analyze the effect of chemotherapeutic drugs and specific kinase inhibitors, in combination with the death receptor ligand tumor necrosis factor related apoptosis inducing ligand (TRAIL), on overcoming TRAIL resistance in hepatocellular carcinoma (HCC) and to study the efficacy of agonistic TRAIL antibodies, as well as the commitment of antiapoptotic BCL-2 proteins, in TRAIL induced apoptosis. METHODS: Surface expression of TRAIL receptors (TRAIL-R1-4) and expression levels of the antiapoptotic BCL-2 proteins MCL-1 and BCL-x(L) were analyzed by flow cytometry and Western blotting, respectively. Knock-down of MCL 1 and BCL-x(L) was performed by transfecting specific small interfering RNAs. HCC cells were treated with kinase inhibitors and chemotherapeutic drugs. Apoptosis induction and cell viability were analyzed via flow cytometry and 3-(4,5 Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. RESULTS: TRAIL-R1 and -R2 were profoundly expressed on the HCC cell lines Huh7 and Hep-G2. However, treatment of Huh7 and Hep-G2 with TRAIL and agonistic antibodies only induced minor apoptosis rates. Apoptosis resistance towards TRAIL could be considerably reduced by adding the chemotherapeutic drugs 5-fluorouracil and doxorubicin as well as the kinase inhibitors LY294002 [inhibition of phosphoinositol-3-kinase (PI3K)], AG1478 (epidermal growth factor receptor kinase), PD98059 (MEK1), rapamycin (mammalian target of rapamycin) and the multi-kinase inhibitor Sorafenib. Furthermore, the antiapoptotic BCL-2 proteins MCL-1 and BCL-x(L) play a major role in TRAIL resistance: knock-down by RNA interference increased TRAIL induced apoptosis of HCC cells. Additionally, knock-down of MCL-1 and BCL-x(L) led to a significant sensitization of HCC cells towards inhibition of both c-Jun N-terminal kinase and PI3K. CONCLUSION: Our data identify the blockage of survival kinases, combination with chemotherapeutic drugs and targeting of antiapoptotic BCL-2 proteins as promising ways to overcome TRAIL resistance in HCC. PMID- 20014457 TI - Diarrhoea-predominant irritable bowel syndrome distinguishable by 16S rRNA gene phylotype quantification. AB - AIM: To study whether selected bacterial 16S ribosomal RNA (rRNA) gene phylotypes are capable of distinguishing irritable bowel syndrome (IBS). METHODS: The faecal microbiota of twenty volunteers with IBS, subdivided into eight diarrhoea predominant (IBS-D), eight constipation-predominant (IBS-C) and four mixed symptom-subtype (IBS-M) IBS patients, and fifteen control subjects, were analysed at three time-points with a set of fourteen quantitative real-time polymerase chain reaction assays. All assays targeted 16S rRNA gene phylotypes putatively associated with IBS, based on 16S rRNA gene library sequence analysis. The target phylotypes were affiliated with Actinobacteria, Bacteroidetes and Firmicutes. Eight of the target phylotypes had less than 95% similarity to cultured bacterial species according to their 16S rRNA gene sequence. The data analyses were made with repeated-measures ANCOVA-type modelling of the data and principle component analysis (PCA) with linear mixed-effects models applied to the principal component scores. RESULTS: Bacterial phylotypes Clostridium cocleatum 88%, Clostridium thermosuccinogenes 85%, Coprobacillus catenaformis 91%, Ruminococcus bromii-like, Ruminococcus torques 91%, and R. torques 93% were detected from all samples analysed. A multivariate analysis of the relative quantities of all 14 bacterial 16S rRNA gene phylotypes suggested that the intestinal microbiota of the IBS-D patients differed from other sample groups. The PCA on the first principal component (PC1), explaining 30.36% of the observed variation in the IBS D patient group, was significantly altered from all other sample groups (IBS-D vs control, P = 0.01; IBS-D vs IBS-M, P = 0.00; IBS-D vs IBS-C, P = 0.05). Significant differences were also observed in the levels of distinct phylotypes using relative values in proportion to the total amount of bacteria. A phylotype with 85% similarity to C. thermosuccinogenes was quantified in significantly different quantities among the IBS-D and control subjects (-4.08 +/- 0.90 vs 3.33 +/- 1.16, P = 0.04) and IBS-D and IBS-M subjects (-4.08 +/- 0.90 vs -3.08 +/ 1.38, P = 0.05). Furthermore, a phylotype with 94% similarity to R. torques was more prevalent in IBS-D patients' intestinal microbiota than in that of control subjects (-2.43 +/- 1.49 vs -4.02 +/- 1.63, P = 0.01). A phylotype with 93% similarity to R. torques was associated with control samples when compared with IBS-M (-2.41 +/- 0.53 vs -2.92 +/- 0.56, P = 0.00). Additionally, a R. bromii like phylotype was associated with IBS-C patients in comparison to control subjects (-1.61 +/- 1.83 vs -3.69 +/- 2.42, P = 0.01). All of the above mentioned phylotype specific alterations were independent of the effect of time. CONCLUSION: Significant phylotype level alterations in the intestinal microbiotas of IBS patients were observed, further emphasizing the possible contribution of the gastrointestinal microbiota in IBS. PMID- 20014459 TI - Education-based approach to addressing non-evidence-based practice in preventing NSAID-associated gastrointestinal complications. AB - AIM: To evaluate an evidence-based educational program for improving strategies for prevention of nonsteroidal anti-inflammatory drug (NSAID)-associated gastrointestinal (GI) complications. METHODS: Four hundred and fifty-six specialists replied to a questionnaire that covered issues related to NSAID induced adverse effects. They also collected data from their last five consecutive patients before and after they had attended an evidence-based seminar on GI prevention strategies. RESULTS: Four hundred and forty-one of 456 specialists (96.7%) participated in the survey, and 382 (83.7%) in the education based study that recorded data from 3728 patients. The specialists overestimated the risk of GI complications with NSAIDs, underestimated the GI safety profile of coxibs, but were aware of the risk factors and of the current prevention strategies. Proton pump inhibitors were co-prescribed with NSAIDs in > 80% of patients with and without risk factors. The educational program had little impact on prescribing habits. CONCLUSION: Specialists are informed of advances in NSAID associated adverse effects and have high rates of GI-prevention therapy. Our educational program did not alter these rates. PMID- 20014460 TI - Hyperphosphatemia after sodium phosphate laxatives in low risk patients: prospective study. AB - AIM: To establish the frequency of hyperphosphatemia following the administration of sodium phosphate laxatives in low-risk patients. METHODS: One hundred consecutive ASA I-II individuals aged 35-74 years, who were undergoing colonic cleansing with oral sodium phosphate (OSP) before colonoscopy were recruited for this prospective study. EXCLUSION CRITERIA: congestive heart failure, chronic kidney disease, diabetes, liver cirrhosis, intestinal obstruction, decreased bowel motility, increased bowel permeability, and hyperparathyroidism. The day before colonoscopy, all the participants entered a 24-h period of diet that consisted of 4 L of clear fluids with sugar or honey and 90 mL (60 g) of OSP in two 45-mL doses, 5 h apart. Serum phosphate was measured before and after the administration of the laxative. RESULTS: The main demographic data (mean +/- SD) were: age, 58.9 +/- 8.4 years; height, 163.8 +/- 8.6 cm; weight, 71 +/- 13 kg; body mass index, 26 +/- 4; women, 66%. Serum phosphate increased from 3.74 +/- 0.56 to 5.58 +/- 1.1 mg/dL, which surpassed the normal value (2.5-4.5 mg/dL) in 87% of the patients. The highest serum phosphate was 9.6 mg/dL. Urea and creatinine remained within normal limits. Post-treatment OSP serum phosphate concentration correlated inversely with glomerular filtration rate (P < 0.007, R(2) = 0.0755), total body water (P < 0.001, R(2) = 0.156) and weight (P < 0.013, R(2) = 0.0635). CONCLUSION: In low-risk, well-hydrated patients, the standard dose of OSP-laxative-induced hyperphosphatemia is related to body weight. PMID- 20014461 TI - Carcinoma of the middle bile duct: is bile duct segmental resection appropriate? AB - AIM: To compare survival between bile duct segmental resection (BDSR) and pancreaticoduodenectomy (PD) for treating distal bile duct cancers. METHODS: Retrospective analysis was conducted for 45 patients in a BDSR group and for 149 patients in a PD group. RESULTS: The T-stage (P < 0.001), lymph node invasion (P = 0.010) and tumor differentiation (P = 0.005) were significant prognostic factors in the BDSR group. The 3- and 5-year overall survival rates for the BDSR group and PD group were 51.7% and 36.6%, respectively and 46.0% and 38.1%, respectively (P = 0.099). The BDSR group and PD group did not show any significant difference in survival when this was adjusted for the TNM stage. The 3- and 5-year survival rates were: stage Ia [BDSR (100.0% and 100.0%) vs PD (76.9% and 68.4%) (P = 0.226)]; stage Ib [BDSR (55.8% and 32.6%) vs PD (59.3% and 59.3%) (P = 0.942)]; stage IIb [BDSR (19.2% and 19.2%) vs PD (31.9% and 14.2%) (P = 0.669)]. CONCLUSION: BDSR can be justified as an alternative radical operation for patients with middle bile duct in selected patients with no adjacent organ invasion and resection margin is negative. PMID- 20014462 TI - Percutaneous transgastric computed tomography-guided biopsy of the pancreas using large needles. AB - AIM: To assess the safety, yield and clinical utility of percutaneous transgastric computed tomography (CT)-guided biopsy of pancreatic tumor using large needles, in selected patients. METHODS: We reviewed 34 CT-guided biopsies in patients with pancreas mass, of whom 24 (71%) had a direct path to the mass without passing through a major organ. The needle passed through the liver in one case (3%). Nine passes (26%) were made through the stomach. These nine transgastric biopsies which used a coaxial technique (i.e. a 17-gauge coaxial introducer needle and an 18-gauge biopsy needle) were the basis of this study. Immediate and late follow-up CT images to detect complications were obtained. RESULTS: Tumor tissues were obtained in nine pancreatic biopsies, and histologic specimens for diagnosis were obtained in all cases. One patient, who had a rare sarcomatoid carcinoma, received a second biopsy. One patient had a complication of transient pneumoperitoneum but no subjective complaints. An immediate imaging study and clinical follow-up detected neither hemorrhage nor peritonitis. No delayed procedure-related complication was seen during the survival period of our patients. CONCLUSION: Pancreatic biopsy can be obtained by a transgastric route using a large needle as an alternative method, without complications of peritonitis or bleeding. PMID- 20014463 TI - Clinicopathological and prognostic analysis of 429 patients with intrahepatic cholangiocarcinoma. AB - AIM: To understand the clinicopathological characteristics and treatment selections and improve survival and provide valuable information for patients with intrahepatic cholangiocarcinoma (ICC). METHODS: We retrospectively evaluated 5311 liver cancer patients who received resection between October 1999 and December 2003. Of these, 429 (8.1%) patients were diagnosed with ICC, and their clinicopathological, surgical, and survival characteristics were analyzed. RESULTS: Upper abdominal discomfort or pain (65.0%), no symptoms (12.1%), and hypodynamia (8.2%) were the major causes for medical attention. Laboratory tests showed 198 (46.4%) patients were HBsAg positive, 90 (21.3%) had alpha-fetoprotein > 20 microg/L, 50 (11.9%) carcinoembryonic antigen > 10 microg/L, and 242 (57.5%) carbohydrate antigen 19-9 (CA19-9) > 37 U/mL. Survival data was available for 329 (76.7%) patients and their mean survival time was 12.4 mo. The overall survival of the patients with R0, R1 resection and punching exploration were 18.3, 6.6 and 5.6 mo, respectively. Additionally, CA19-9 > 37 U/mL was associated with lymph node metastases, but inversely associated with cirrhosis. Multivariate analysis indicated that radical resection, lymph node metastases, macroscopic tumor thrombi and size, and CA19-9 were associated with prognosis. CONCLUSION: Surgical radical resection is still the most effective means to cure ICC. Certain laboratory tests (such as CA19-9) can effectively predict the survival of the patients with ICC. PMID- 20014464 TI - Neoadjuvant chemoradiotherapy for resectable esophageal carcinoma: a meta analysis. AB - AIM: To compare neoadjuvant chemoradiotherapy and surgery with surgery alone for resectable esophageal carcinoma. METHODS: We used MEDLINE and EMBASE databases to identify eligible studies and manual searches were done to ensure no studies were missed. Trial validity assessment was performed and a trial quality score was assigned. RESULTS: Eleven randomized controlled trials (RCTs) including 1308 patients were selected. Neoadjuvant chemoradiotherapy significantly improved the overall survival compared with surgery alone. Odds ratio (OR) [95% confidence interval (CI), P value], expressed as neoadjuvant chemoradiotherapy and surgery vs surgery alone, was 1.28 (1.01-1.64, P = 0.05) for 1-year survival, 1.78 (1.20 2.66, P = 0.004) for 3-year survival, and 1.46 (1.07-1.99, P = 0.02) for 5-year survival. Postoperative mortality increased in patients treated by neoadjuvant chemoradiotherapy (OR: 1.68, 95% CI: 1.03-2.73, P = 0.04), but incidence of postoperative complications was similar in two groups (OR: 1.14, 95% CI: 0.88 1.49, P = 0.32). Neoadjuvant chemoradiotherapy lowered the local-regional cancer recurrence (OR: 0.64, 95% CI: 0.41-0.99, P = 0.04), but incidence of distant cancer recurrence was similar (OR: 0.94, 95% CI: 0.68-1.31, P = 0.73). Histological subgroup analysis indicated that esophageal squamous cell carcinoma did not benefit from neoadjuvant chemoradiotherapy, OR (95% CI, P value) was 1.16 (0.85-1.57, P = 0.34) for 1-year survival, 1.34 (0.98-1.82, P = 0.07) for 3-year survival and 1.41 (0.98-2.02, P = 0.06) for 5-year survival. CONCLUSION: Neoadjuvant chemoradiotherapy can raise the survival rate of patients with esophageal adenocarcinoma. PMID- 20014465 TI - Effect of severe acute pancreatitis on pharmacokinetics of Da-Cheng-Qi Decoction components. AB - AIM: To investigate the effect of severe acute pancreatitis (SAP) on pharmacokinetics of Da-Cheng-Qi Decoction (DCQD) components in rats. METHODS: Rats were divided into SAP group and sham-operation group as a control group (n = 6). Rhein, chrysophanol, rheochrysidin, magnolol, hesperidin and naringin in DCQD were quantified in rat serum by high performance liquid chromatography tandem mass spectrometry for studying their pharmacokinetics. RESULTS: Early absorption of each DCQD component was tended to degrade in SAP group after treatment with DCQD by gavage. The C(max) (chrysophanol, P = 0.0059; rheochrysidin, P = 0.0288; magnolol, P = 0.0487; hesperidin, P = 0.0277; naringin, P = 0.0023) and AUC (rhein, P = 0.0186; chrysophanol, P = 0.0013; magnolol, P = 0.001; hesperidin, P = 0.0081; naringin, P = 0.0272) of DCQD component were obviously lower in SAP group than in control group. The T(1/2alpha) of chrysophanol and rheochrysidin (P = 0.0467 and 0.0005, respectively) and T(max) of chrysophanol and rheochrysidin (P = 0.0101 and 0.0037, respectively) lasted longer in SAP group than in control group. CONCLUSION: SAP can significantly impact the absorption of DCQD components in rats and their pharmacokinetic parameters. PMID- 20014466 TI - Complete response to radiation therapy of orbital metastasis from hepatocellular carcinoma. AB - The incidence of hepatocellular carcinoma (HCC) is increasing in the United States, and 50%-75% of patients with HCC will develop metastatic disease. Orbital metastases from HCC are extremely rare. We report the case of a 52-year-old male with known metastatic HCC, who presented with severe proptosis and diplopia. An orbital mass was identified on magnetic resonance imaging (MRI) and confirmed to have hypermetabolic activity on positron emission tomography/computed tomography. He received a palliative course of external beam radiation therapy to the right orbit. Intensity modulated radiation therapy (IMRT) was used to allow sparing of critical normal tissues in close proximity to the tumor. One month after completion of IMRT to 58 Gray in 30 fractions delivered over 6 wk, the patient had a complete clinical, radiologic (MRI) and symptomatic response. The patient continues to have local control in the orbit 1.7 years after therapy completion. All critical normal structures were kept below the tolerance dose using IMRT, and no toxicities were observed. PMID- 20014467 TI - Superior mesenteric artery syndrome in a diabetic patient with acute weight loss. AB - Superior mesenteric artery (SMA) syndrome is an uncommon disease resulting compression of the third portion of the duodenum from the superior mesenteric artery. This disease shares many common manifestations with diabetic gastroparesis, including postprandial fullness, nausea, vomiting, and bloating. Therefore, it is often overlooked in diabetic patients. Here, we report a 41-year old man with poorly controlled diabetic mellitus who developed SMA syndrome due to rapid weight loss. The diagnosis was confirmed by computed tomography and an upper gastrointestinal series. His condition improved after parenteral nutrient, strict sugar control, and gradual weight gain. PMID- 20014468 TI - Aortoesophageal fistula: a case misdiagnosed as esophageal polyp. AB - Aortoesophageal fistula (AEF) is a rare and fatal disorder. It is also a life threatening cause of massive upper gastrointestinal hemorrhage. Thoracic aortic aneurysm is the most common cause of AEF. Management of a patient with this disorder requires rapid diagnosis and immediate intervention, which is considered the best way to save the patient's life. We report a case of AEF misdiagnosed as esophageal polyp. PMID- 20014469 TI - Frequency of alcohol and smoking cessation counseling in hepatitis C patients among internists and gastroenterologists. AB - Given the overwhelming evidence that both alcohol consumption and smoking accelerate the progression of hepatitis C virus (HCV)-induced liver disease, we evaluated the frequency of alcohol and smoking counseling of patients with HCV induced liver disease by their primary care internists and gastroenterologists. One hundred and twenty-three medical records of consecutive patients with HCV induced liver disease referred by an internist to a gastroenterologist for its management were reviewed. Patient gender, race, history of and counseling against alcohol and tobacco use by a physician and a gastroenterologist were obtained. A database was created using Microsoft Excel. There were 105 African-Americans, 12 Caucasians and six patients of other races/ethnicities. Forty-six (37%) patients were daily tobacco users and 34 (28%) patients were daily alcohol consumers. There was a statistically significant difference in the frequencies of alcohol (P = 0.0002) and smoking cessation (P = 0.0022) between gastroenterologists and internists. This study reveals that internists and gastroenterologists, alike, inadequately counsel patients with hepatitis C about tobacco and alcohol use. PMID- 20014470 TI - Analysis of mesenchymal stem cell differentiation in vitro using classification association rule mining. AB - In this paper, data mining is used to analyze the data on the differentiation of mammalian Mesenchymal Stem Cells (MSCs), aiming at discovering known and hidden rules governing MSC differentiation, following the establishment of a web-based public database containing experimental data on the MSC proliferation and differentiation. To this effect, a web-based public interactive database comprising the key parameters which influence the fate and destiny of mammalian MSCs has been constructed and analyzed using Classification Association Rule Mining (CARM) as a data-mining technique. The results show that the proposed approach is technically feasible and performs well with respect to the accuracy of (classification) prediction. Key rules mined from the constructed MSC database are consistent with experimental observations, indicating the validity of the method developed and the first step in the application of data mining to the study of MSCs. PMID- 20014471 TI - On the asymmetry of the residue compositions of the binding sites on protein surfaces. AB - By screening all the ligand binding sites in the Protein Data Bank, we have found that while it is geometrically possible that a loop, formed from a protein chain with residues ZYX, would "impersonate" another chain-loop with residues XYZ by a simple twisting of either the loop or the bound ligand, it almost never happens. This fact is rather surprising, and implies a notable asymmetry, since (i) loops in the folded proteins sometimes can be flexible enough to be twisted, but (ii) ligands are almost always extremely mobile before binding to the protein, therefore they can turn around and bind to residue-sequence ZYX as well. Data availability: The supplementary Table 3 lists the appearances of the residue sequences and their inverses in the binding sites of the whole PDB, and is available at http://www.worldscient.com/jbcb/. PMID- 20014472 TI - Dimension reduction of microarray gene expression data: the accelerated failure time model. AB - The construction of the components of Partial Least Squares (PLS) is based on the maximization of the covariance/correlation between linear combinations of the predictors and the response. However, the usual Pearson correlation is influenced by outliers in the response or in the predictors. To cope with outliers, we replace the Pearson correlation with the Spearman rank correlation in the optimization criteria of PLS. The rank-based method of PLS is insensitive to outlying values in both the predictors and response, and incorporates the censoring information by using an approach of Nguyen and Rocke (2004) and two approaches of reweighting and mean imputation of Datta et al. (2007). The performance of the rank-based approaches of PLS, denoted by Rank-based Modified Partial Least Squares (RMPLS), Rank-based Reweighted Partial Least Squares (RRWPLS), and Rank-based Mean-Imputation Partial Least Squares (RMIPLS), is investigated in a simulation study and on four real datasets, under an Accelerated Failure Time (AFT) model, against their un-ranked counterparts, and several other dimension reduction techniques. The results indicate that RMPLS is a better dimension reduction method than other variants of PLS as well as other considered methods in terms of the minimized cross-validation error of fit and the mean squared error of fit in the presence of outliers in the response, and is comparable to other variants of PLS in the absence of outliers. Supplementary Materials are available at http://www.worldscinet.com/jbcb/ PMID- 20014473 TI - A probabilistic framework to improve microrna target prediction by incorporating proteomics data. AB - Due to the difficulties in identifying microRNA (miRNA) targets experimentally in a high-throughput manner, several computational approaches have been proposed. To this date, most leading algorithms are based on sequence information alone. However, there has been limited overlap between these predictions, implying high false-positive rates, which underlines the limitation of sequence-based approaches. Considering the repressive nature of miRNAs at the mRNA translational level, here we describe a probabilistic model to make predictions by combining sequence complementarity, miRNA expression level, and protein abundance. Our underlying assumption is that, given sequence complementarity between a miRNA and its putative mRNA targets, the miRNA expression level should be high and the protein abundance of the mRNA should be low. Having identified a set of confident predictions, we then built a second probabilistic model to trace back to the mRNA expression of the confident targets to investigate the mechanisms of the miRNA mediated post-transcriptional regulation. Our results suggest that translational repression (which has no effect on mRNA level), instead of mRNA degradation, is the dominant mechanism in miRNA regulation. This observation explained the previously observed discordant correlation between mRNA expression and protein abundance. PMID- 20014474 TI - Sirius PSB: a generic system for analysis of biological sequences. AB - Computational tools are essential components of modern biological research. For example, BLAST searches can be used to identify related proteins based on sequence homology, or when a new genome is sequenced, prediction models can be used to annotate functional sites such as transcription start sites, translation initiation sites and polyadenylation sites and to predict protein localization. Here we present Sirius Prediction Systems Builder (PSB), a new computational tool for sequence analysis, classification and searching. Sirius PSB has four main operations: (1) Building a classifier, (2) Deploying a classifier, (3) Search for proteins similar to query proteins, (4) Preliminary and post-prediction analysis. Sirius PSB supports all these operations via a simple and interactive graphical user interface. Besides being a convenient tool, Sirius PSB has also introduced two novelties in sequence analysis. Firstly, genetic algorithm is used to identify interesting features in the feature space. Secondly, instead of the conventional method of searching for similar proteins via sequence similarity, we introduced searching via features' similarity. To demonstrate the capabilities of Sirius PSB, we have built two prediction models - one for the recognition of Arabidopsis polyadenylation sites and another for the subcellular localization of proteins. Both systems are competitive against current state-of-the-art models based on evaluation of public datasets. More notably, the time and effort required to build each model is greatly reduced with the assistance of Sirius PSB. Furthermore, we show that under certain conditions when BLAST is unable to find related proteins, Sirius PSB can identify functionally related proteins based on their biophysical similarities. Sirius PSB and its related supplements are available at: http://compbio.ddns.comp.nus.edu.sg/~sirius. PMID- 20014475 TI - In silico screening of protein-protein interactions with all-to-all rigid docking and clustering: an application to pathway analysis. AB - We propose a computational screening system of protein-protein interactions using tertiary structure data. Our system combines all-to-all protein docking and clustering to find interacting protein pairs. We tuned our prediction system by applying various parameters and clustering algorithms and succeeded in outperforming previous methods. This method was also applied to a biological pathway estimation problem to show its use in network level analysis. The structural data were collected from the Protein Data Bank, PDB. Then all-to-all docking among target protein structures was conducted using a conventional protein-protein docking software package, ZDOCK. The highest-ranked 2000 decoys were clustered based on structural similarity among the predicted docking forms. The features of generated clusters were analyzed to estimate the biological relevance of protein-protein interactions. Our system achieves a best F-measure value of 0.43 when applied to a subset of general protein-protein docking benchmark data. The same system was applied to protein data in a bacterial chemotaxis pathway, utilizing essentially the same parameter set as the benchmark data. We obtained 0.45 for the F-measure value. The proposed approach to computational PPI detection is a promising methodology for mediating between structural studies and systems biology by utilizing cumulative protein structure data for pathway analysis. PMID- 20014476 TI - Inference of gene regulatory networks using boolean-network inference methods. AB - The modeling of genetic networks especially from microarray and related data has become an important aspect of the biosciences. This review takes a fresh look at a specific family of models used for constructing genetic networks, the so-called Boolean networks. The review outlines the various different types of Boolean network developed to date, from the original Random Boolean Network to the current Probabilistic Boolean Network. In addition, some of the different inference methods available to infer these genetic networks are also examined. Where possible, particular attention is paid to input requirements as well as the efficiency, advantages and drawbacks of each method. Though the Boolean network model is one of many models available for network inference today, it is well established and remains a topic of considerable interest in the field of genetic network inference. Hybrids of Boolean networks with other approaches may well be the way forward in inferring the most informative networks. PMID- 20014477 TI - Exploring the protein landscape in ramachandran space: it's not just psi-phi. AB - Most methods for the structural comparison of proteins utilize molecular coordinates in the three-dimensional physical space. Recently, a group has presented an elegant novel approach based on the characterization of protein shape in terms of backbone torsion angles. They have demonstrated considerable success in direct comparisons with other techniques, and their method lends itself to rapid screening of structural information from rapidly growing databases. We think that the torsion angle approach can be further strengthened by refining the distance notion that forms the basis of the computational scheme. In particular, we are suggesting to compute the distance along the path that minimizes the transition cost between aligned pairs of angles and therefore likely provides a more meaningful representation of distances between points in Ramachandran space. PMID- 20014478 TI - [Advances in application of deep brain stimulation in treatment of neuropsychological diseases]. AB - Deep brain stimulation has drawn more and more concerns as a method to treat neuropsychological diseases. Compared with surgery and other methods using electrical stimulation, deep brain stimulation has advantages of clear targets, high selectivity, reversibility, titratability and non-ablation. A large body of clinical trials has shown that deep brain stimulation targeting various brain structures is able to alleviate the symptoms of Parkinson's disease, epilepsy, chronic pain and depression that are intractable with medicines and other methods, with few complications or side effects. Deep brain stimulation is now emerging as a promising approach for the treatment of resistant neuropsychological diseases. PMID- 20014479 TI - [Effects of clobenpropit and histidine on reinstatement of morphine-induced conditioned place preference]. AB - OBJECTIVE: To investigate the effects of clobenpropit and histidine on reinstatement of morphine-induced conditioned place preference (CPP) in rats. METHODS: The persistence, extinction and reinstatement of morphine-induced CPP were established.In clobenpropit group three different doses of clobenpropit (2, 5 and 10 microg/rat, i.c.v.) were administered 15 min after morphine (1 mg/kg, i.p.) was injected. In histidine group histidine (100, 200, 500 mg/kg) was given 1 h prior to morphine treatment (1 mg/kg i.p). RESULT: The CPP was reinstated by priming injection of 1 mg/kg morphine. Clobenpropit (5, 10 microg/rat) significantly inhabited the reinstatement by a priming dose of morphine-induced CPP compared with the morphine control group; histidine (100, 200, 500 mg/kg) significantly inhibited the reinstatement in a dose-dependent manner. CONCLUSION: Clobenpropit and histidine inhibit the revival of morphine-induced CPP in a dose dependent manner, indicating that endogenous histamine may inhibit relapse of morphine to some extent. PMID- 20014480 TI - [Intranasal administration of low dosage recombinant human erythropoietin inhibits seizure in rats]. AB - OBJECTIVE: To investigate the effect of intranasal administration of low dosage recombinant human erythropoietin (r-HuEPO) on seizure in rats. METHODS: After intranasal or intraperitoneal administration of r-HuEPO, the behavioral and electroencephalographic changes were observed in pentylenetetrazol (PTZ) and maximal electroshock (MES) induced seizure or electrical amygdaloid-kindled seizure of rats. RESULT: Intranasal administration of low dosage r-HuEPO increased the seizure latency, and decreased the seizure grade and duration, and the number of convulsive episodes in PTZ induced seizure, with the most potential dosage of 2.4 IU. Intraperitoneal administration of r-HuEPO (3 000, 4 000 IU/kg) only decreased the seizure duration and number of convulsive episodes. The seizure grade, forelimb or hindlimb extension duration were decreased in MES induced seizure by intranasal administration of 2.4 IU r-HuEPO. In addition, intranasal administration of 2.4 IU r-HuEPO decreased the seizure grade, generalized seizure duration and afterdischarge in electrical amygdaloid-kindled rats stimulated with generalized seizure threshold. CONCLUSION: Intranasal administration of low dosage r-HuEPO can inhibit the seizure in rats. PMID- 20014481 TI - [Delayed neuronal degeneration after intracerebral hemorrhage: the role of iron]. AB - OBJECTIVE: To investigate the occurrence of delayed neuronal degeneration, activation of microglia and nuclear factor-kappa B after rat intracerebral hemorrhage (ICH) and the possible role of iron. METHODS: ICH model was induced by infusion of autologous whole blood into the right basal ganglia. To evaluate the possible role of iron on delayed neuron loss, an iron model by injection of FeCl(2) into hippocampus was also set up. Degeneration of neurons and the activation of microglia and NF-kappa B were detected. RESULT: Both whole blood and iron caused neuron degeneration for at least 14 days were revealed by Fluoro jade C staining. Consistently, activated microglia and NF-kappa B positive cells were also observed in the peri-hematoma area and the ipsilateral hippocampus. CONCLUSION: The iron may participate in the delayed neuron injury followed ICH; the activated microglia and NF-kappa B may be involved in the process of delayed neuronal injury. PMID- 20014482 TI - [Effect of endogenous histamine on ischemic preconditioning induced cerebral ischemic tolerance]. AB - OBJECTIVE: To investigate the effect of endogenous histamine on ischemic preconditioning induced cerebral ischemic tolerance in rats. METHODS: Wild-type (WT) mice and histidine decarboxylase knock-out (HDC-KO) mice were preconditioned by bilateral carotid artery occlusion (BCCAO) for 6, 10,or 14 min and reperfused for 48 h,then subjected to permanent BCCAO and the survival time of WT and HDC-KO mice subjected to permanent BCCAO was observed. Histamine levels in the hypothalamus, hippocampus, striatum and cortex at 0.5 h,5 h or 48 h after 10 min BCCAO were determined with high-performance liquid chromatography. RESULT: Ten minutes ischemic preconditioning significantly prolonged the survival time of WT mice subjected to permanent BCCAO. However,in HDC-KO mice, the ischemic tolerance was not induced with 10 min preconditioning. The histamine levels at 0.5 h or 48 h increased after 10 min preconditioning, but not at 5 h. CONCLUSION: Endogenous histamine in brain may be an essential mediator in ischemic preconditioning induced cerebral ischemic tolerance. PMID- 20014483 TI - [Construction and identification of eukaryotic expression vector of rat GPR17 gene]. AB - OBJECTIVE: To construct the eukaryotic expression vector of rat GPR17 (rGPR17) cDNA,and to identify its function in HEK293 cells. METHODS: Total RNA was extracted from rat brain tissue; full-length GPR17 cDNA was prepared by RT-PCR, and cloned into pcDNA3.1(+) plasmid. The recombinant plasmid was converted into E.coli DH5alpha and confirmed by PCR, double enzyme digestion analysis and DNA sequencing. The recombinant plasmid pcDNA3.1(+)-rGPR17 was transiently transfected into HEK293 cells using Lipofectamin 2000. Expression of rGPR17 gene was confirmed by RT-PCR and immunofluorescence staining. The exogenous LTD(4) enhanced intracellular calcium was measured using Fluo-4. RESULT: RT-PCR, double enzyme digestion analysis and sequencing showed that the rGPR17 gene was cloned into recombinant vector, and the recombinant rGPR17 was expressed after transfection in HKE293 cells. LTD(4) increased intracellular calcium release in the transfected HEK293 cells. CONCLUSION: The eukaryotic expression vector of rGPR17 cDNA has been constructed; it is functionally expressed in HEK293 cells. This work provides a basis for further research of the GPR17 receptor and its antagonists. PMID- 20014484 TI - [Preparation and identification of polyclonal antibody against cysteinyl leukotriene receptor 2]. AB - OBJECTIVE: To prepare and identify a polyclonal antibody against cysteinyl leukotriene receptor (CysLT(2)receptor). METHODS: Rabbits were immunized with KLH coupled CysLT(2) receptor peptide to prepare the polyclonal antibody (pAb). The titer of the pAb in rabbit plasma was detected by indirect ELISA, and the specificity of the pAb was tested by antigen blockade. The tissue distribution of CysLT(2) receptor was detected by Western blot and immunohistochemistry with the prepared pAb. RESULT: The pAb showed a titer higher than 1/1047296, and was specific to CysLT(2) receptor, without cross-reaction with the antigens of CysLT(1) receptor and GPR17. A higher expression of CysLT(2) receptor in kidney, brain and lung of rats and mice was detected by Western blot analysis using the prepared pAb. The molecular weight of CysLT(2) receptor protein was about 40 kD. Immunohistochemical examination showed that CysLT(2) receptor was expressed mainly in the neuron, and partly in astrocytes in rat brain. CONCLUSION: The prepared CysLT(2) receptor pAb has high sensitivity and specificity, and can be used in Western blot and immunohistochemistry. PMID- 20014485 TI - [Method for screening cysteinyl leukotriene receptor 2 antagonists and preliminary screening of compounds]. AB - OBJECTIVE: To establish a method for screening cysteinyl leukotriene receptor 2 (CysLT(2)) antagonists and to preliminarily screen a series of synthetic compounds. METHODS: Rat glioma cell line (C6 cells) highly expressing CysLT(2) receptor was used. Intracellular calcium concentration was measured after stimulation with the agonist LTD(4),which was used to screen compounds with antagonist activity for CysLT(2) receptor. Bay u9773, a CysLT1/CysLT(2) receptor non-selective antagonist, and AP-100984, a CysLT(2) receptor antagonist, were used as control. RESULT: PT-PCR showed a higher expression of CysLT(2) receptor in C6 cells. LTD(4) at 1 mumol/L significantly increased intracellular calcium in C6 cells; the maximal effect was about 37.5% of ATP, a positive stimulus.LTD(4) induced increase of intracellular calcium was blocked by CysLT(2) receptor antagonists, but not by CysLT(1) receptor antagonists. Among the synthetic compounds, D(XW-)1,2,13,23,29 and 30 inhibited LTD(4)-induced increase of intracellular calcium. CONCLUSION: LTD(4)-induced change in intracellular calcium in C6 cells can be used as a screening method for CysLT(2) receptor antagonists. The compounds, D(XW-)1,2,13,23,29 and 30, possess antagonist activity for CysLT(2) receptor. PMID- 20014486 TI - [Reliability and validity of a Chinese version of the Quality of Life in Epilepsy Inventory (QOLIE-31-P)]. AB - OBJECTIVE: To develop and validate a Chinese version of the Patient-Weighted Quality of Life in Epilepsy Questionnaire (QOLIE-31-P). METHODS: The original English version of the QOLIE-31-P was translated into the Chinese language. The inventory was then completed by 200 adult patients with epilepsy; and 49 patients also completed the scale twice within three weeks. Test retest, internal consistency reliabilities, construct validity, and some influential factors for quality of life in adults with epilepsy were assessed. RESULT: Test retest reliability (Pearson's correlation coefficient) for the Chinese version of the QOLIE-31-P ranged from 0.725 to 0.912 (P<0.001), and the internal consistency reliability (Cronbach's alpha coefficient) ranged from 0.627 to 0.898. Factor analysis showed that there were seven factors, which explained the total variance for 64.9%. The coefficient of seizure frequency with the quality of life was 0.81(P<0.05). The QOLIE-31-P score of the patients with tonic clonic seizure was 55.7 +/-16.6, and that of other seizure type was 61.4 +/-18.7(t=-2.568, P<0.05). CONCLUSION: The psychometric properties of the Chinese version of the QOLIE-31-P have satisfactory reliability and validity, and can be applied to assess quality of life in Chinese adult patients with epilepsy. PMID- 20014487 TI - [Change of JNK MAPK and its influence on cardiocyte apoptosis in ischemic postconditioning]. AB - OBJECTIVE: To test whether postconditioning could inhibit the expression of phospho-JNK (P-JNK) mitogen activated protein kinase (MAPK) and study its relation to apoptosis of cardiocyte. METHODS: Sixty rats were randomly divided into six groups: sham, reperfusion injury (R/I), postconditioning (Post), SP600125 (I_JNK), anisomycin and postconditioning (Ani+Post) and anisomycin (Ani) groups. After acute myocardial infarction was induced in rats, placebo solution (DMSO), SP600125 (6 mg/kg) or anisomycin (2 mg/kg) was injected through jugular vein 5 min before reperfusion; 6 h later 3 rats of each group were executed and the hearts were separated to measure the signaling molecules (phospho-JNK, TNF alpha, Caspase-8, Bcl-2/Bax, cytochrome-c). Twenty-two hours later hemodynamic data were measured in the left rats, and then blood samples were taken to determine serum markers of cardiac damage, and hearts were separated to measure the infarction area and cardiocyte apoptosis. RESULT: Postconditioning improved +/-DP/DTmax of left ventricle, limited infarct area, relieved apoptosis and necrosis of cardiocytes, and inhibited the expression of P-JNK (1.12 +/-0.21 Compared with 1.90 +/-0.32, P<0.05). At the same time the levels of TNFalpha Caspase-8, Bax and Cyt-c were lower in Post group than those in R/I group, but Bcl-2 expression levels were higher. I_JNK group presented the similar protection effect of postconditioning [TUNEL index: (6.23 +/-2.43)% Compared with (18.22 +/ 5.10)%, P<0.05; Infarct area: (23.44 +/-6.34)% Compared with (42.31 +/-8.21)%, P<0.05]. On the other hand, Ani+Post group partially lost cardioprotection effect [TUNEL index: (14.12 +/-2.00)% Compared with (18.22 +/-5.10)%,P>0.05; Infarct area: (35.27 +/-5.28)% Compared with (42.31+/-8.21)%,P>0.05], because of the activation of JNK MAPK. CONCLUSION: Postconditioning can inhibit phosphorylation of JNK MAPK, which attenuates cardiocyte apoptosis by both extrinsic and mitochondria pathway. PMID- 20014488 TI - [Relationship between oxidative stress and beta cell lipoapoptosis in rats]. AB - OBJECTIVE: To investigate the effect of beta cell lipoapoptosis after long term high-fat feeding in rats, and to investigate the relationship between oxidative stress, gene expression and beta cell lipoapoptosis. METHODS: Forty-one SD male rats were randomly divided into 2 groups: high-fat diet group (HF group) and control group (NC group). At the end of 28 weeks, the levels of malondialdehyde (MDA) and glutamylcysteinylglycine (GSH) in plasma and pancreatic tissue,the early-phase insulin secretion in beta cells, the beta cell apoptosis (TUNEL technology) and the uncoupling protein 2 (UCP2) gene expression in islets were measured. RESULT: The concentrations of MDA both in plasma and pancreatic tissue were higher in HF group than those in NC group.In contrast, The contents of GSH both in plasma and pancreatic tissue were lower in HF group. Insulin secretion response to glucose load was significantly decreased in HF group (3.0 fold Compared with 5.7 fold, P<0.01). Blood glucose levels at 3 min, 5 min and 10 min during IVGTT were significantly higher in HF group than those in NC group (P<0.05). The frequency of beta cell apoptosis was increased by 40.0% in HF group (P<0.01). The gene expression of UCP2 in islets was increased by 22.4% in HF group (P<0.01). CONCLUSION: The frequency of beta cell apoptosis in high-fat feeding rats is affected by oxidative stress, which results in increasing UCP2 gene expression. PMID- 20014489 TI - [An in vitro hepatic steatosis cell model for study of non-alcoholic fatty liver disease]. AB - OBJECTIVE: To establish an in vitro cell model to investigate hepatic steatosis of non-alcoholic fatty liver disease. METHODS: HepG2 cells cultured in MEM containing 10 % fetal bovine serum were divided into control group and model group. At 7 0%-80 % confluency, HepG2 cells in model group were exposed to a long chain mixture of free fatty acids (oleate and palmitate) for 24 h, cells in control group were subject to fresh medium. Lipid droplets were observed with oil red O stain and electron microscope, triglyceride and malonaldehyde were detected by respective assay kits. RESULT: A large number of lipid droplet were detected in model HepG2 cells; the level of triglyceride increased. However,malonaldehyde did not increase significantly compared with control group. CONCLUSION: A large number of lipid droplet were detected in model HepG2 cells; the level of triglyceride increased. However, malonaldehyde did not increase significantly compared with control group. PMID- 20014490 TI - [Relationship of polymorphism of SG13S114A/T in ALOX5AP gene with atherosclerotic cerebral infarction]. AB - OBJECTIVE: To investigate the relationship of the polymorphism of SG13S114A/T in ALOX5AP gene with atherosclerotic cerebral infarction (ACI). METHODS: By polymerase chain reaction and restriction fragment length polymorphism, polymorphism of SG13S114A/T in ALOX5AP gene in 412 cases with ACI and 368 non-ACI controls were analyzed. RESULT: There were no statistically significant differences in the ALOX5AP gene SG13S114 AA genotype and A allele frequencies between ACI group and control group (P>0.05). CONCLUSION: The results do not support genotype SG13S114 A allele as the risk gene for ACI.control group. PMID- 20014491 TI - [Evaluation of left ventricular systolic asynchrony in patients with uremic myocardiopathy using tissue synchronization imaging]. AB - OBJECTIVE: To evaluate the left ventricular systolic asynchrony in patients with uremic myocardiopathy (UM) using tissue synchronization imaging (TSI). METHODS: Ultrasound system with TSI and Q-analyze software were used. Thirty-five patients with UM were enrolled in the study,and thirty normal subjects were included as the control group. RESULT: The total and mean time to peak velocity (Tc) corrected by the heart rate of all segments in UM group were longer than those in the control group (P<0.05), and the time to peak velocity of most segments in UM group was also longer (P<0.05). Delayed time to peak velocity was found in 175 (175/420) segments in UM group and left ventricular systolic asynchrony was detected in 65.7% (23/35). CONCLUSION: TSI can detect the ventricular systolic asynchrony in patients with uremic myocardiopathy and provide reliable parameters for clinical management. PMID- 20014492 TI - [Recent advances in treatment of glial scar-induced inhibition of nerve regeneration]. AB - Glial scar formed by central nervous system (CNS) injury is the main inhibitory barrier of nerve regeneration. How to promote axonal regeneration after injury,how to accelerate neural network reconstruction and how to improve brain function recovery have become a hot problem to be solved in the field of neuroscience. This article focuses on the recent advances of therapeutic strategies for axonal regeneration. PMID- 20014493 TI - [VEGF and central nervous system diseases]. AB - Vascular endothelial growth factor (VEGF or VEGF-A) is a hypoxia induced angiogenic growth factor that is potent in neurotrophy,neuroprotection, anti apoptosis and cell proliferation. Recent reports suggest that VEGF is related to many central nervous system diseases, such as cerebral ischemic disease, Alzheimer's disease and Parkinson's disease. Further study of the relationship between VEGF and central nervous system diseases,and investigation of VEGF related drugs will shed light on a new way for treatment of central nervous system diseases. PMID- 20014494 TI - [PICK1 is associated with central nervous system diseases]. AB - PICK1 (protein interacting with C kinase 1) contains a PDZ (PSD-95/Dlg/ZO1) domain and a BAR (Bin/amphiphysin/Rvs) domain. Via the PDZ domain, PICK1 interacts directly with more than 40 proteins. Among these interacting proteins, some are important for physiological and pathophysiological processes of central nervous system. In this review, recent findings about how PICK1 is associated with central nervous system diseases are summarize. PMID- 20014495 TI - Strength training, Part II: from theory to practice. PMID- 20014496 TI - Premature heart disease. PMID- 20014497 TI - Calling all men: cell phones and sperm. PMID- 20014498 TI - On call. I've been tracking my PSA ever since I turned 50, 11 years ago. Since I just moved from St. Louis to Denver, my next test will be done in a new lab. Will the change in labs affect my levels? PMID- 20014499 TI - Emerging issues. Study says racial HIV disparities an 'indictment' of U.S. response. PMID- 20014500 TI - Incarceration. HIV/AIDS epidemic in prisons is problematic, researchers say. PMID- 20014502 TI - Sex offender not required to submit to involuntary HIV test. PMID- 20014501 TI - SSI. ALJ did not identify evidence to support denial of benefits. PMID- 20014503 TI - Data trends. Report reveals startling statistics about blacks with HIV. PMID- 20014504 TI - Comparative dental morphology. Preface. PMID- 20014505 TI - The Reverend Thomas Hincks FRS (1818-1899): taxonomist of Bryozoa and Hydrozoa. AB - Thomas Hincks was born 15 July 1818 in Exeter, England. He attended Manchester New College, York, from 1833 to 1839, and received a B.A. from the University of London in 1840. In 1839 he commenced a 30-year career as a cleric, and served with distinction at Unitarian chapels in Ireland and England. Meanwhile, he enthusiastically pursued interests in natural history. A breakdown in his health and permanent voice impairment during 1867-68 while at Mill Hill Chapel, Leeds, forced him reluctantly to resign from active ministry in 1869. He moved to Taunton and later to Clifton, and devoted much of the rest of his life to natural history. Hincks was elected a Fellow of the Royal Society of London in 1872 for noteworthy contributions to natural history. Foremost among his publications in science were "A history of the British hydroid zoophytes" (1868) and "A history of the British marine Polyzoa" (1880). Hincks named 24 families, 52 genera and 360 species and subspecies of invertebrates, mostly Bryozoa and Hydrozoa. Hincks died 25 January 1899 in Clifton, and was buried in Leeds. His important bryozoan and hydroid collections are in the Natural History Museum, London. At least six genera and 13 species of invertebrates are named in his honour. PMID- 20014506 TI - The fate of the bird specimens from Cook's voyages possessed by Sir Joseph Banks. AB - Joseph Banks possessed the greater part of the zoological specimens collected on James Cook's three voyages round the world (1768-1780). In early 1792, Banks divided his zoological collection between John Hunter and the British Museum. It is probable that those donations together comprised most of the zoological specimens then in the possession of Banks, including such bird specimens as remained of those that had been collected by himself and Daniel Solander on Cook's first voyage, and those that had been presented to him from Cook's second and third voyages. The bird specimens included in the Banks donations of 1792 became part of a series of transactions during the succeeding 53 years which involved the British Museum, the Royal College of Surgeons of England, and William Bullock. It is a great pity that, of the extensive collection of bird specimens from Cook's voyages once possessed by Banks, only two are known with any certainty to survive. PMID- 20014507 TI - Charles Wesley Hargitt (1852-1927): American educator and cnidarian biologist. AB - Charles Wesley Hargitt was born near Lawrenceburg, Indiana, USA, and died at Syracuse, New York. After a brief career as a Methodist Episcopal Minister, he carried out graduate studies in biology at Illinois Wesleyan University and Ohio University. He served briefly on the faculty at Moores Hill College and later at Miami University of Ohio before receiving an appointment at Syracuse University. Hargitt spent 36 years at Syracuse, and for 21 years was a trustee of the Marine Biological Laboratory, Woods Hole, Massachusetts. His research encompassed animal behaviour, cell biology, development, ecology, natural history, and taxonomy, as well as education, eugenics, and theology, and he wrote or contributed to more than 100 publications in science. Approximately half of these were on Cnidaria, with 41 of them on Hydrozoa. His most important works in hydrozoan taxonomy were on species of the Woods Hole region, the Philippines, and south China. Hargitt was author of three genera and 48 species and subspecies ascribed to Hydrozoa, seven species of Anthozoa, and one species of Cubozoa. Four species of hydroids are named in his honour. PMID- 20014508 TI - Botany and zoology in the late seventeenth-century Philippines: the work of Georg Josef Camel SJ (1661-1706). AB - Georg Josef Camel (1661-1706) went to the Spanish colony of the Philippine Islands as a Jesuit lay brother in 1687, and he remained there until his death. Throughout his time in the Philippines, Camel collected examples of the flora and fauna, which he drew and described in detail. This paper offers an overview of his life, his publications and the Camel manuscripts, drawings and specimens that are preserved among the Sloane Manuscripts in the British Library and in the Sloane Herbarium at the Natural History Museum, London. It also discusses Camel's links and exchanges with scientifically minded plant collectors and botanists in London, Madras and Batavia. Among those with whom Camel corresponded were John Ray, James Petiver, and the Dutch physician Willem Ten Rhijne. PMID- 20014509 TI - Bute's "Botanical tables": dictated by nature. AB - In the final years of his life, after a long and turbulent political career, John Stuart, third Earl of Bute, was at last free to indulge in one of his passions: botany. The publication of Linnaeus's "Systema naturae" in 1735 threw the botanical world into disarray and academic argument raged throughout Europe. The production of the "Botanical tables" (1785) was an ambitious project to explain Bute's individual view of Linnaeus's system of taxonomy and was particularly composed for the "Fair Sex". Twelve volumes were published privately and presented to family, royalty and botanical colleagues across Europe. The "Botanical tables" were illustrated by the renowned botanical artist, John Miller. The illustrations are both aesthetically pleasing and scientifically correct. In this paper we consider the circumstances of the production of the "Botanical tables" and explore how the original sets of this publication and original material have been dispersed. PMID- 20014510 TI - [States and societies facing collectives and individuals: Sweden during the 19th century]. PMID- 20014511 TI - [An honor or a duty? The second pinkas of the Swarzedz mutual aid society, 1772 1809]. PMID- 20014512 TI - Yellow flashing light on EMRs. PMID- 20014513 TI - CRS. PMID- 20014514 TI - So who's counting? PMID- 20014515 TI - Limited-time opportunity. Arkansas hopes to advance HIT through federal funding. PMID- 20014516 TI - Health care quality and disparities and the health care provider: what we can do. PMID- 20014517 TI - Case report of a 30.8 pound cystosarcoma phyllodes of breast. AB - This is a case report and literature review of a 30.8 pound cystosarcoma phyllodes tumor resected in a 54-year-old female. Starting at age 19, the patient had noted a small breast tumor that continued to grow over the years. Pathology reported a low-grade malignant phyllodes tumor with no nodal involvement. Surgical resection with clear margins still remains the main treatment for this tumor. Radiation may be helpful in those with positive margins, a tumor greater than 4 cm, and those with reoccurrence. PMID- 20014518 TI - Initial infectious disease screening evaluation of the international adoptee. AB - International adoptees are consistently exposed to unbelievable living conditions prior to being adopted that, coupled with malnourished states, substandard medical assessment and care, and unreliable immunizations, place them at significant risk for infectious diseases. In this second in a three-part series of topics related to the care of international adoptees, we discuss recommended infectious disease screening tests that can be provided by the primary care physician once the child arrives in the United States. PMID- 20014519 TI - Legally speaking: EMTALA and pandemics. PMID- 20014520 TI - Big-five personality, social worldviews, and ideological attitudes: further tests of a dual process cognitive-motivational model. AB - In this study, we extended the Dual Process Model of Ideology and Prejudice by incorporating the Five-Factor Model of Personality (N = 924). Disagreeable people tended to view the social world as competitive, which in turn predicted heightened motivations for group-based dominance and superiority (Social Dominance Orientation or SDO), whereas people low in Openness to Experience and high in Conscientiousness directly expressed heightened security-cohesion motivations (Right-Wing Authoritarianism or RWA). Other personality dimensions were weakly associated with RWA, and these effects were mediated by dangerous worldview. Multiple distinct aspects of personality predict SDO and RWA both directly and indirectly through worldviews, but we found little evidence for the possibility that personality alters the extent to which worldviews (once formed) predict SDO and RWA. PMID- 20014521 TI - Back to the Consideration of Future Consequences Scale: time to reconsider? AB - The Consideration of Future Consequences (CFC) Scale is a measure of the extent to which individuals consider and are influenced by the distant outcomes of current behavior. In this study, the authors conducted factor analysis to investigate the factor structure of the 12-item CFC Scale. The authors found evidence for a multiple factor solution including one completely present-oriented factor consisting of all 7 present-oriented items, and one or two future-oriented factors consisting of the remaining future-oriented items. Further evidence indicated that the present-oriented factor and the 12-item CFC Scale perform similarly in terms of internal consistency and convergent validity. The structure and content of the future-oriented factor(s) is unclear. From the findings, the authors raise questions regarding the construct validity of the CFC Scale, the interpretation of its results, and the usefulness of the CFC scale in its current form in applied research. PMID- 20014522 TI - Self-ratings of physical attractiveness in a competitive context: when males are more sensitive to self-perceptions than females. AB - The authors investigated sex differences in the ratings of physical attractiveness in a competitive context. Participants in an Ultimatum Game experiment offered to split a sum of money with their opponents who could either accept or reject the offers; subsequently, physical attractiveness ratings (both self-ratings and of the other) were obtained. The authors found that male participants rated themselves higher on physical attractiveness when facing male opponents than when facing female ones; there was no such difference for the female participants. Furthermore, male participants' self-ratings of physical attractiveness were higher than the ratings provided by their corresponding male opponents. The authors discuss these findings using the tenets of evolutionary psychology pertaining to as male intra-sexual rivalry in competitive contexts. PMID- 20014523 TI - A web replication of Snyder, Decker, and Bersheid's (1977) experiment on the self fulfilling nature of social stereotypes. AB - Authors instructed two groups of male participants to respond to a web personal advertisement that had a photograph of either an attractive or unattractive woman. In the physically attractiveness condition, it was found that the message was longer and contained few spelling and grammatical mistakes than in the condition where the male's participant believe that the personal advertisement came from an unattractive woman. Two undergradutates females who were unaware of the experimental conditions judged that they had more probability to respond to the male's message in the physically attractiveness condition. PMID- 20014528 TI - [Which care plan for a wound?]. PMID- 20014527 TI - [Osteoporosis and pain]. PMID- 20014529 TI - [Improving the care of multiple sclerosis patients]. PMID- 20014530 TI - [Multiple sclerosis physiopathology]. PMID- 20014531 TI - [Multiple sclerosis, from the first signs to the diagnostic consultation]. PMID- 20014532 TI - [Multiple sclerosis, the therapeutic arsenal of today and tomorrow]. PMID- 20014533 TI - [The nurse's role in managing the multiple sclerosis patient]. PMID- 20014534 TI - [Rehabilitation and functional recovery of patients with multiple sclerosis]. PMID- 20014535 TI - [Multiple sclerosis in children]. PMID- 20014536 TI - [A history of multiple sclerosis]. PMID- 20014537 TI - [Association for Research on Multiple Sclerosis, to support research and inform the public]. PMID- 20014538 TI - [A health network for patients with multiple sclerosis]. PMID- 20014539 TI - [Dynamic research shows significant therapeutic advances]. PMID- 20014540 TI - [9/10. Organizing and coordinating nursing interventions (competence 9)]. PMID- 20014541 TI - [2/5. The nurse's role in managing pain]. PMID- 20014542 TI - Lessons learned in implementing a 24/7 public health call center in response to H1N1 in the state of New Jersey. AB - OBJECTIVE: The purpose of this article was to collect, examine, and report the data obtained in response to opening a 24/7 Call Center in response to the H1N1 influenza outbreak in the State of New Jersey in the Spring of 2009. DESIGN: Data log sheets were collected and analyzed based on phone calls received into the State of New Jersey H1N1 Call Center during the initial response to the H1N1 public health emergency from April to May 2009. Data were stratified to examine the types of calls received, where they originated, and the types of organizations/agencies that needed guidance/information during the initial response to the H1N1 public health emergency. Additionally, lessons learned from this operational response were documented. RESULTS: 3,855 calls were received and analyzed during the first 8 days of commencing the H1N1 Call Center. Signs and symptoms were the main category of questions asked, representing 31.2 percent of the call volume. Of the 3,855 calls, 216 (5.6 percent) were from agencies, such as healthcare institutions. Multiple lessons learned were documented from a planning and operations perspective. CONCLUSIONS: Communication to the general public is paramount to ensure accurate information is being conveyed during a public health response. The lessons learned from this operation are currently being utilized in response to the H1N1 influenza outbreak during the Fall of 2009. PMID- 20014543 TI - Resources and constraints for addressing ethical issues in medical humanitarian work: experiences of expatriate healthcare professionals. AB - OBJECTIVE: International nongovernmental organizations frequently provide emergency assistance in settings where armed conflict or natural disaster overwhelm the capacity of local and national agencies to respond to health and related needs of affected communities. Healthcare practice in humanitarian settings presents distinct clinical, logistical, and ethical challenges for clinicians and differs in important ways from clinical practice in the home countries of expatriate healthcare professionals. The aim of this research was to examine the moral experience of healthcare professionals who participate in humanitarian relief work. DESIGN: I conducted a qualitative research study using interpretive description methodology. PARTICIPANTS: Fifteen Canadian healthcare professionals and three human resource or field coordination officers for nongovernmental organizations were interviewed. RESULTS: In this article, I present findings related to expatriate healthcare professionals' experiences of resources and constraints for addressing ethical issues in humanitarian crises. Resources for ethics deliberation and reflection include the following: opportunities for discussion; accessing and understanding local perspectives; access to outside perspectives; attitudes, such as humility, open-mindedness, and reflexivity; and development of good moral "reflexes." Constraints for deliberation and reflection relate to three domains: individual considerations, contextual features of humanitarian relief work, and local team and project factors. CONCLUSION: These findings illuminate the complex nature of ethical reflection, deliberation, and decision-making in humanitarian healthcare practice. Healthcare professionals and relief organizations should seek to build upon resources for addressing ethical issues. When possible, they should minimize the impact of features that function as constraints. PMID- 20014544 TI - Cyclone Nargis in Myanmar: lessons for public health preparedness for cyclones. AB - Recent natural disasters such as the 2004 tsunami, 2008 Sichuan earthquake, and the 2008 Myanmar cyclone have killed more than 100,000 people each. Mortality and morbidity associated with natural disasters are a growing concern, especially because extreme climate events are likely to get increasingly frequent. The authors comment on Cyclone Nargis, claiming an extraordinarily high death toll during its devastating track through the Irrawaddy delta in Myanmar on May 2, 2008 and analyze how and why its mortality pattern differs from other typical postdisaster situations. Underlying factors and preconditions are described and the specificity of the Myanmese context is presented. This leads to lessons how excess mortality can be reduced in future high-ranked cyclones, whose recurrence in this region will only be a matter of time. PMID- 20014545 TI - Disaster preparedness: are retired physicians willing to help? AB - OBJECTIVE: To identify the proportion of retired physicians belonging to a state wide professional association who would be willing to volunteer in the event of a disaster. METHODS: A paper-based, self-administered questionnaire sent to all physicians listed as retired members of the Washington State Medical Association (WSMA). The main questions included whether subjects would be willing to volunteer during a disaster, which tasks they would be most willing to perform, and whether they would be willing to participate in disaster preparedness training. RESULTS: A total of 2,443 surveys were mailed, 2,274 arrived at their destination (169 were undeliverable), and 1,447 were returned (response rate 64 percent). Fifty-four percent of respondents reported they would be willing to perform healthcare tasks during a disaster and 24 percent of respondents said they would possibly be willing to help. Tasks retired physicians were most willing to assist with included minor wound care (85 percent), vaccine administration (74 percent), and starting intravenous lines (71 percent). Fewer respondents indicated willingness to assist with community education (60 percent) or staffing ambulatory clinics (48 percent). Seventy-eight percent indicated they would attend disaster preparedness training. CONCLUSIONS: Healthcare facilities must be prepared to cope with staffing shortages in the event of a disaster and volunteers such as retired physicians could fill crucial roles in a medical response plan. The majority of retired physicians surveyed would be willing to participate. They would be most willing to perform well-defined tasks directly related to patient care. Most would be willing to participate in preparatory training. PMID- 20014546 TI - H1N1 and institutions of higher education. AB - OBJECTIVE: Institutions of Higher Education (IHE) have been preparing for the likely resurgence of Influenza A (H1N1) virus this Fall. Amongst the multitude of factors affecting their preparatory efforts, medical considerations and evidence serve to provide the foundation for many planning decisions. DESIGN: The authors reviewed the relevant medical literature for evidence of effective measures to mitigate the consequences of H1N1. Evidence was reviewed as it pertains to IHE. The authors opted to focus on vaccination, antiviral medications, masks, hand washing, environmental cleaning, and isolation and quarantine. RESULTS: Despite the limited evidence found for the IHE setting, recommendations were made to encourage vaccination, deemphasize the role of antivirals in most IHE students, and provide surgical masks for ill students, as they may leave their living environment, while simultaneously stressing self isolation without quarantine. Additionally, frequent hand washing and high traffic fomite cleaning should be encouraged. CONCLUSION: Preparation for pandemic influenza in the IHE context is very complex and all decisions should be based on the best evidence available. PMID- 20014547 TI - Hospital preparedness in Nebraska: a pandemic influenza survey. AB - Because hospital response to pandemic influenza is a critical component of preparedness, the authors performed a survey of hospitals in Nebraska to assess pandemic influenza preparedness. An anonymous 15-question survey was sent electronically to all 85 hospitals in Nebraska in November 2008 and 48 hospitals (56.5 percent) responded. A large majority of Nebraska hospitals have a pandemic influenza plan (n = 41/48, 85.4 percent), and have begun stockpiling materials (n = 44/47, 93.6 percent). Most have established contact with local public health (n = 42//47, 89.4 percent), and in fact reported that their communication with public health has improved as a result of pandemic influenza planning (n = 39/46, 84.8 percent). This survey illuminates current progress and areas for potential improvement in pandemic influenza planning by Nebraska hospitals. PMID- 20014548 TI - Immunostaining as a diagnostic aid in cytopathologic study of upper urinary tract urothelial carcinoma. AB - OBJECTIVE: To evaluate preoperative diagnosis of low-grade urothelial carcinoma (LGUC) and urothelial neoplasms of unknown malignant potential (UMP UN) of the upper urinary tract (UUT) and its role in disease management, especially in the context of nephron-sparing treatment possibilities. STUDY DESIGN: Wash and brush ureteral specimens of LGUC/UMP UN of the UUT with histopathologic correlation were retrieved at our institution for 7 years and studied along with 7 ureteral specimens from nonneoplastic ureteral lesions. RESULTS: Of 30 specimens from 25 LGUC/UMP UN, 5 were negative for tumor cells and 3 showed cytologic atypia. The remaining 22 contained tumor cells with characteristic features of urothelial carcinoma, including hard and soft criteria. The 4 hard criteria included branching stromal cores, dyshesive cell networks, 3-dimensional papillary clusters with stromal core and atypia associated with CK20-positive cells. The 2 soft criteria were hypercellularity and atypia in CK20-negative cells. All LGUC/UMP UN of the UUT were associated with at least 1 hard criterion or both soft criteria. CONCLUSION: Branching stromal cores, 3-dimensional papillary clusters, dyshesive cell networks and CK20-positive atypia immunostaining appear specific for LGUC/UMP UN of the UUT but are seen in few cases. Combined soft and hard criteria will increase sensitivity to 83%. PMID- 20014549 TI - Diagnostic utility of MOC-31, HBME-1 and MOC-31 mRNA in distinguishing between carcinoma cells and reactive mesothelial cells in pleural effusions. AB - OBJECTIVE: To evaluate the individual and combined diagnostic utility of MOC-31, HBME-1 and MOC-31mRNA in the differentiation of adenocarcinoma cells from reactive mesothelial cells in pleural effusions. STUDY DESIGN: To subject the cells from 293 pleural effusions to immunocytochemical staining for MOC-31 and HBME-1 and reverse transcriptase polymerase chain reaction for the expression of MOC-31. RESULTS: MOC-31 and MOC-31mRNA expression were significantly higher in the cancer cell group and in 3 subgrouped as adenocarcinoma, squamous cell carcinoma and small cell lung carcinoma than in the mesothelial cell group, whereas HBME-1 expression was obviously lower in those (p < 0.01). In single use, MOC-31mRNA had the highest sensitivity (90.8%) and accuracy (91.5%), whereas MOC 31 had the highest specificity (100%). When combinations of markers were evaluated together, MOC-31mRNA and HBME-1 gave a high diagnostic performance: sensitivity of 96.7% and accuracy of 95.2%, respectively. CONCLUSION: The detection of MOC-31, HBME-1 and MOC-31-mRNA is helpful in distinguishing between cancer and reactive mesothelial cells in pleural effusions. MOC-31mRNA could be useful to the diagnosis of pleural micrometastasis. PMID- 20014550 TI - The adjunctive value of CD10 immunostaining on cell block preparations in pelvic endometriosis. AB - OBJECTIVE: To evaluate the utility of CD10 immunostaining on cell block preparations from pelvic washing cytology material obtained during laparoscopy in cases of pelvic endometriosis (PE). STUDY DESIGN: Six premenopausal women presenting with ovarian masses underwent a laparoscopic procedure. In addition to routine cytospin preparations, cell blocks were prepared using the formalin fixed, paraffin-embedded method from the pelvic washings in each case. Immunocytochemistry using monoclonal antibodies for cytokeratin (CK) 7, CK20, CD10, estrogen receptor (ER), progesterone receptor (PR) and CA125 was performed on sections from cell blocks using the streptavidin-biotin peroxidase complex. RESULTS: Clusters of epithelial cells (3 cases) and stromal cells (2 cases) were seen in a background rich in hemosiderin-laden macrophages and poorly preserved mesothelial cells on cytospin smears. CD10 and CK7 were consistently positive in the stromal and epithelial components, respectively, in all cases on cell block sections. Variable staining reactions were noted for ER, PR and CA125. CONCLUSION: CD10 immunostaining is a useful ancillary method in the diagnosis of endometriosis. Its routine use on cell block preparations from pelvic washings in women undergoing a laparoscopic procedure is recommended. PMID- 20014551 TI - Application of cytomorphologic features and immunocytochemistry to cervicovaginal smears. A scoring system to distinguish between endometrial and endocervical adenocarcinoma. AB - OBJECTIVE: To apply cytomorphologic features and a limited panel of immunocytochemistry to cervicovaginal smears to distinguish between primary endocervical adenocarcinoma (ECA) and primary endometrial adenocarcinoma (EMC). STUDY DESIGN: The study was a retrospective analysis of 35 cases, of which 21 were of primary endometrial origin and 14 were of primary endocervical origin. In all cases included in the study, the site of origin of adenocarcinoma was confirmed by examination of the surgical specimen. Twenty-four cytomorphologic characteristics and a limited panel of antibodies (estrogen receptor, carcinoembryonic antigen and vimentin) were applied to conventional cervicovaginal smears to ascertain whether a combination of these would be sufficiently distinctive to allow reliable distinction between ECA and EMC. The slides were scored using defined cytomorphologic characteristics and immunocytochemistry. The score was calculated by using the receiver operating curve (ROC). RESULTS: Statistical analysis identified 7 variables that were significant in distinguishing between ECA and EMC: necrosis, altered blood, histiocytes, strips of cells, palisading of cells, pseudorosettes and positive immunocytochemical staining for carcinoembryonic antigen. Using the ROC, a numerical score < 4 was indicative of an EMC and a score > or = 4 of ECA. CONCLUSION: This study found that the proposed scoring system based on these 7 variables could reliably distinguish between ECA and EMC and assist in definitive management of patients. PMID- 20014552 TI - Atypical bare nuclei in liquid-based cervical cytology and their significance. AB - OBJECTIVE: To investigate the potential significance of atypical bare nuclei in ThinPrep (TP) technique (Cytyc Corp., Boxborough, Massachusetts, U.S.A.), liquid based cervical cytology and their possible association with certain epithelial abnormalities. STUDY DESIGN: Four thousand seven hundred thirty-six TP cervical smears were reviewed over a period of 13 months. Cases in which atypical bare nuclei were detected were selected and were further studied for the clinical, cytomorphologic and histologic features associated with each case. Cases with "typical" bland-looking, free nuclei were excluded. RESULTS: Ten cases were found to have atypical bare nuclei in the reviewed TP cervical smears. Nine were found to be associated with high grade squamous intraepithelial lesions (HSILs) and 6 were nonkeratinizing-type HSIL, 5 of which had endocervical involvement. None were associated with low grade squamous intraepithelial lesions (LSILs). One was associated with endometrial serous papillary carcinoma. CONCLUSION: In this study we tried to illustrate the significance of atypical bare nuclei in TP cervical smears. An association of atypical bare nuclei with HSIL, but not with LSIL, was observed. Attempts were made to explain this association and to suggest the potential management implications for TP cervical smears with such atypical bare nuclei. PMID- 20014553 TI - Does providing previous results change the accuracy of cervical cytology? AB - OBJECTIVE: To determine whether providing previous cytology and histology findings alters the accuracy of conventional cervical cytology reading or changes reading times. STUDY DESIGN: Each of 9 cytologists read 9 batches of 8 routinely referred Pap smears (total, 648 slides), with history (H) and without history (NH), at an interval of no less than 4 weeks. Each batch was read blind to the result of reading under the other strategy and to histology. Histologic cervical intraepithelial neoplasia 2 or more severe was the reference standard. Accuracy of reading was assessed across all thresholds using receiver operating characteristic (ROC) curves and by sensitivity and specificity at a cytology threshold of possible low grade squamous intraepithelial lesion (consistent with atypical squamous cells of undermined significance). RESULTS: Areas under the ROC curve, sensitivities and specificities were similar if read with or without history, except for 1 reader for whom reading with history increased the area under the ROC curve from 0.716 to 0.833 (increase of 0.117, p = 0.017) and the sensitivity from 0.57 to 0.79 (increase of 0.22, p = 0.014), without any significant change in specificity. Accuracy varied between subgroups defined by age and by the severity and timing of previous abnormalities, but the results of the comparison of accuracy in H and NH did not vary by subgroup. Mean reading times were 8.2 (H) and 7.9 (NH) minutes per slide, a difference of 0.34 minutes (p = 0.083). Differences in mean batch times (H-NH) between readers ranged from 0.08 to 1.0 minutes, the largest difference being for the reader whose accuracy increased. CONCLUSION: An accurate history might improve accuracy for some cytologists. PMID- 20014554 TI - A quality control study on cytotechnologist-cytopathologist concordance and its relationship to the number of dots on the slide. AB - OBJECTIVE: To study cytotechnologist (CT)-cytopathologist (CP) concordance for evaluating individual CTs' performance and for quality assurance and educational feedback. STUDY DESIGN: The interpretations of individual CTs were compared with the final interpretations (according to the 2001 Bethesda System) of the CP. Concordance percentages and kappa values were calculated for each CT and correlated with the numbers of dots on each slide, years of experience and percentage of work hours devoted to cytology. RESULTS: A total of 10,453 Pap tests were screened by 9 CTs during one year, out of which 993 (9.5%) were referred to one CP for a final interpretation. Mean concordance between the aggregate CT interpretations and those of the CP was 65.5%. Five CTs had good concordance, 3 had moderately good concordance, and one had surprisingly poor concordance that contrasted with good subjective impressions. No correlation was found between concordance and the average number of dots per slide, screening experience in cervicovaginal cytology or percentage of work hours devoted to cytology. CONCLUSION: Monitoring CT-CP concordance rates can unveil performance issues not detected by subjective impressions. An excessive number of dots per slide may not reflect poor diagnostic precision so much as a lack of confidence in interpretation. PMID- 20014555 TI - Diagnostic efficacy of conventional as compared to liquid-based cytology in thyroid lesions: evaluation of 10,360 fine needle aspiration cytology cases. AB - OBJECTIVE: To evaluate the efficacy of thyroid fine needle aspiration cytology (FNAC) processed by liquid-based cytology (LBC) as compared to conventional smears (CS) in 10,360 consecutive cases in 3 reference periods. STUDY DESIGN: In 1997-1998, 2,394 FNACs had CS; in 2001-2002, 3,463 had both CS and LBC; and in 2004-2005, 4,503 had only LBC. Three arbitrary parameters of efficacy were chosen to compare the two different methods: rate of inadequacy (IR), rate of indeterminacy (INDR) and rate of malignancy (MR). In the same biennia 6,184 thyroidectomy specimens were examined. RESULTS: IR was, respectively, 8.4% in 1997-1998, 18.4% in 2001-2002 and 13.3% in 2004-2005. MR was respectively 2.2%, 2.5% and 2.2%. INDR was 16.6% in 1997-1998, 23.3% in 2001-2002 and 15.9% in 2004 2005. In each biennium, respectively, 16.9%, 27.6% and 28% of malignant neoplasms were histologically diagnosed. CONCLUSION: LBC was used as an alternative method in the cytologic diagnosis for the preoperative selection of thyroid neoplasms, decreasing indeterminate diagnoses without MR changes. PMID- 20014556 TI - Fine needle aspiration cytology of granulomatous mastitis with special emphasis on microbiologic correlation. AB - OBJECTIVE: To describe the cytomorphology of granulomatous mastitis (GM) and to evaluate the causative agents involved, proven on the basis of microbiologic culture results and polymerase chain reaction (PCR) studies. STUDY DESIGN: We retrospectively reviewed the inflammatory breast aspirates reported at King Abdul Aziz Specialist Hospital and Al Hada Armed Forces Hospital, Taif, Kingdom of Saudi Arabia, from January 2000 until March 2008. The pathology reports, clinical information, slides, microbiologic culture results and tuberculosis (TB)/PCR were reviewed. Cases of inflammatory carcinoma or duct ectasia were excluded. RESULTS: A total of 49 cases of inflammatory breast aspirates were identified, of which 15 cases were GM. The microbiologic cultures of all 15 cases were available. Six of 15 cases had positive culture for Brucella melitenses, and 2 cases were positive for TB, which was further confirmed by TB/PCR. The fungal cultures were negative in all the cases. Seven of 15 cases were classified as idiopathic GM. CONCLUSION: Fine needle aspiration cytology was found to be a useful tool in the early diagnosis of GM and its classification if performed by a pathologist as the material can be submitted for culture. The definitive diagnosis can be established by a combination of the cytomorphologic features and microbiologic studies. PMID- 20014557 TI - Cytopathologic analysis of paratracheal masses: a study of 737 cases with clinicoradiologic correlation. AB - OBJECTIVE: To analyze the cytopathologic findings of a series of paratracheal space (PTS) masses in the context of clinicoradiologic correlation. STUDY DESIGN: Retrospective review of our cytopathology files revealed 131 cases of PTS lesions in a 14-year period (1991-2005). Cytologic material was obtained under radiologic guidance. Radiologic findings, clinical data and subsequently performed tissue biopsies were reviewed and correlated. RESULTS: Radiologic imaging disclosed masses in the PTS ranging from 1 to 7 cm. Of the 131 cases, 103 (79%) were deemed diagnostic. Of these, 41 (40%) revealed nonneoplastic lesions, and 62 (60%) yielded malignant neoplasms. Nonneoplastic entities included: 31 (73%) hyperplastic lymph nodes and 10 (24%) sarcoidosis. Of the malignant cases, 45 (73%) were metastatic tumors: adenocarcinoma (ACA) 19, small cell carcinoma 12, squamous cell carcinoma (SQCC) 11 and other tumors, from lung 34, esophagus 4 and other sites. Malignant neoplasms from local spread included lung non-small cell carcinoma 6, SQCC 3 and ACA 3, papillary thyroid carcinoma 3 and other 2. CONCLUSION: Fine needle aspiration (FNA) of PTS has a high diagnostic yield (79%) with a sensitivity of 97% and specificity of 100%. The most common diagnosis is a malignant tumor (60%), with metastatic carcinoma (73%) the most common neoplasm (lung ACA the most common primary source). The most common benign entity is a hyperplastic lymph node (24%). Ancillary studies (immunoctyochemistry, fluorescence in situ hybridization and electron microscopy) were helpful and provided definitive diagnosis in 30% of the initially nondiagnostic FNA samples. PMID- 20014558 TI - Cytologic features of trichoblastoma in fine needle aspiration biopsies. AB - OBJECTIVE: To review the cytologic features of trichoblastoma in order to define criteria that may aid in identification of these tumors at the time of aspiration and allow a definitive diagnosis. STUDY DESIGN: A 58-year-old male presented with a mass lesion on the thigh. On fine needle aspiration, the patient was diagnosed as having a benign skin adnexal tumor. Histology showed the presence of a trichoblastic fibroma, and a retrospective analysis of the cytology was performed. RESULTS: The cytologic features of trichoblastoma resembled a cellular fibroadenoma/phyllodes tumor on aspiration, not previously described in the literature. If the cytomorphology of a skin or subcutaneous aspirate appears to resemble that of a fibroadenoma, the diagnosis of a trichoblastoma should be entertained. Peripheral palisading of nuclei at the edges of the basaloid cell sheets and squamous eddy formation are clues to the diagnosis but may be very focal and could be overlooked. If the tumor occurs in the region of the breast, distinction from a fibroadenoma would be difficult if these additional features were not prominent. CONCLUSION: Knowledge of the cytologic features of trichoblastoma will allow correct management of the patient and prevent misdiagnosis as other benign or malignant tumors. PMID- 20014559 TI - Fine needle aspiration cytology of pilomatrixoma and differential diagnoses. AB - OBJECTIVE: To review the cytologic features of pilomatrixoma to allow a definitive diagnosis at the time of aspiration and correct patient management. STUDY DESIGN: Three patients each presented with a mass lesion and on fine needle aspiration were diagnosed as either having pilomatrixoma or a benign skin adnexal tumor. The diagnosis was confirmed on histology. A retrospective analysis of the cytology was performed. RESULT: The most consistent and predominant features for diagnosis were basaloid cells, anucleate squamous cells and calcification in an inflammatory background, often with giant cells. Only focal evidence of true shadow or "ghost" cells was seen in one case on the Diff-Quik smear. Therefore, lack of shadow cells does not preclude correct diagnosis in the presence of the other relevant cytologic features. If all the major components of pilomatrixoma are present in an aspirate, the diagnosis should not be problematic. However, if one component predominates, this may lead to an erroneous diagnosis of other benign and malignant lesions. CONCLUSION: Knowledge of the cytologic features of pilomatrixoma will allow correct patient diagnosis and management. PMID- 20014560 TI - Low grade fibromyxoid sarcoma: report of a case with epithelioid cell morphology, masquerading as a papillary thyroid carcinoma. AB - BACKGROUND: Low grade fibromyxoid sarcoma (LGFMS) is a rare tumor with a characteristic histologic appearance. Although the spectrum of histologic features that can be seen in LGFMS a well characterized, their description in the cytology literature is lacking. CASE: A 57-year-old male with a past history of Hodgkin's lymphoma presented with 1-year history of dysphagia. Clinical examination and radiologic imaging studies revealed a neck mass suggestive of a thyroid lesion. Fine needle aspiration cytology of the mass revealed moderately cellular smears with several cohesive clusters. Most of the clusters showed cellular crowding with overlapping nuclei. The nuclei ranged from round to oval or elongated shapes with frequent irregular nuclear membranes, powdery to clear nuclear chromatin and inconspicuous nucleoli. Most notably, several intranuclear inclusions and nuclear grooves were noted, leading to a suggestion of a papillary thyroid carcinoma. Histopathologic examination of the mass revealed an LGFMS. CONCLUSION: This report describes a rare case of LGFMS with epithelioid morphology. By virtue of its clinical presentation and cytologic features, the case masqueraded as a papillary thyroid carcinoma and should be added to the list of pitfalls in the diagnosis of papillary thyroid carcinoma. The case also serves to add to the spectrum of morphologic changes that can be seen in LGFMS. PMID- 20014562 TI - Primary mucinous eccrine adenocarcinoma of the scalp: a case report. AB - BACKGROUND: Lesions of primary mucinous carcinoma of the skin present as painless, papular or nodular masses with sizes ranging from 5 mm to 120 mm. Metastatic deposits from undiagnosed visceral and breast adenocarcinoma are virtually indistinguishable microscopically from sweat gland carcinoma and must be considered before a diagnosis of sweat gland carcinoma is made. CASE: A 60 year-old woman presenting with a swelling over the scalp of 1 year's duration was clinically diagnosed to have a sebaceous cyst. Fine needle aspiration cytology of the lesion showed malignant cells in a mucoid background. Histologic examination of the excised mass revealed nests of tumor cells floating in pools of mucin in the dermis. Many of these nests showed tubular lumina. A diagnosis of mucinous eccrine adenocarcinoma was given. CONCLUSION: Primary mucinous adenocarcinoma of the skin is a rare tumor and mimics metastatic adenocarcinoma. The scalp is the second common site for these rumors. Literature search regarding cytologic findings shows our case could be the second case reported so far. PMID- 20014561 TI - Imprint cytologic features in renal cell carcinoma associated with Xp11.2 translocation/TFE3 gene fusion in an adult: a case report. AB - BACKGROUND: Adult-onset renal cell carcinoma (RCC) associated with Xp11.2 translocation/TFE3 gene fusion is a very rare tumor. To date, there are no reports on immunocytochemical study of the primary tumor. We describe such a case that we diagnosed by immunocytochemistry of imprint cytology material. CASE: A 46 year-old man was found to have a mass in the lower pole of the right kidney. Magnetic resonance imaging (MRI) T2-weighted images showed a hypointense area in the tumor, and papillary RCC was suspected. Imprint cytology showed tumor cells that were isolated or arranged in large or small papillary clusters. Irregularly shaped large oval nuclei, finely granular chromatin and a single large nucleolus were noted. Cytoplasm was abundant and admixed with clear and granular eosinophilic patterns and scattered large vacuolated cells. Almost all tumor cells diffusely expressed immunocytochemical reactivity to TFE3 protein. Hyaline nodules were observed in the stroma. Ultrastructurally, neoplastic cells contained rhomboid crystals identical to those of alveolar soft part sarcoma. CONCLUSION: The immunocytochemistry of TFE3 protein may be a powerful tool for accurate diagnosis when RCC associated with Xp11.2 translocation/TFE3 gene fusion is suspected by imprint cytology even in adult-onset cases, and cytotechnologists should accurately recognize cytologic findings of this tumor. PMID- 20014564 TI - Cytologic features and differential diagnosis in a case of extraskeletal mesenchymal chondrosarcoma: a case report. AB - BACKGROUND: Extraskeletal myxoid chondrosarcoma (EMC) is an uncommon tumor. On fine needle aspiration (FNA) it has to be distinguished from other benign and malignant soft tissue lesions. CASE: FNA was done on an 85-year-old man with painful swelling of the forearm. Smears showed fragments comprised of polygonal cells with eccentric nuclei and peripheral fine cytoplasmic vacuoles embedded in a dense, metachromatic matrix. Cell block showed similar cells in a chondromyxoid stroma. The cells were positive for S100 and negative for cytokeratin. CONCLUSION: FNA is a useful tool in the diagnosis of EMC in conjunction with radiology. A preoperative diagnosis can be made due to its distinct cytologic and immunohistochemical features, obviating the need for a biopsy. PMID- 20014563 TI - Ascitic fluid cytology of yolk sac tumor of the ovary: a case report. AB - BACKGROUND: Yolk sac tumor (YST) of the ovary is a rare neoplasm typically affecting children and young women. We describe the cytomorphology of this tumor in ascitic fluid and discuss its differential diagnosis from other neoplasms. CASE: Smear preparations of the ascitic fluid showed a predominance of clusters of malignant cells with vacuolated cytoplasm, mimicking a mucinous adenocarcinoma, and fewer syncytial-like and glandular structures. Hyaline globules were extremely rare. Immunocytochemistry was positive for alpha fetoprotein. CONCLUSION: The diagnosis of YST in ascitic fluid specimens may be challenging and requires the use of immunocytochemistry and the correlation of cytology with the remaining clinical and laboratory data. PMID- 20014565 TI - Fine needle aspiration cytopathology of primary solid adenoid cystic carcinoma of the lung: a case report. AB - BACKGROUND: Adenoid cystic carcinoma (ACC) is a rare, distinctive salivary gland malignant neoplasm arising infrequently as a primary tumor in the lung. Cytomorphology of lung primary ACC has been rarely reported. To the best of our knowledge, cytopathologic features of the solid type of ACC have not been described as a primary lung tumor. CASE: A case of solid-type primary pulmonary ACC arising in the upper segment of the left mainstem bronchus, suggestively diagnosed cytopathologically by bronchoscope-guided fine needle aspiration (FNA), is presented. Cytopathologic differential diagnoses of ACC with other, more common primary neoplasms of the lung are discussed. FNA smears exhibited a normocellular specimen with several large, densely cellular tissue fragments and individual cells, homogeneous and without prominent atypia. Nuclei were round to oval, with finely granular chromatin. Nucleoli were either absent or inconspicious. The nuclear to cytoplasmic ratio was high. Cytoplasm was minimal; chromatin was bland, with rare chromocenters. There was no nuclear molding. Staining the air-dried smears with May-Grunwald-Giemsa stain revealed metachromatic, magenta material not in spherules or cords but as shapeless structures surrounded by tumor cells. CONCLUSION: A preoperative cytopathologic diagnosis of poorly differentiated primary pulmonary ACC can enhance surgical planning and success of tumor resection. PMID- 20014566 TI - The importance of recognizing parasitic organisms in cytologic specimens. PMID- 20014567 TI - Cytologic analysis of the urinary sediment. PMID- 20014568 TI - Clinical significance of reporting benign-appearing endometrial cells in Pap tests in women aged 40 years and over. PMID- 20014569 TI - Diagnosis of pure squamous cell carcinoma of the breast by fine needle aspiration cytology. PMID- 20014570 TI - Acute promyelocytic leukemia with atypical cytologic features. PMID- 20014571 TI - Exploited reefs protected from fishing transform over decades into conservation features otherwise absent from seascapes. AB - Tasmanian reef communities within "no-take" marine protected areas (MPAs) exhibited direct and indirect ecological changes that increasingly manifested over 16 years, eventually transforming into communities not otherwise present in the regional seascape. Data from 14 temperate and subtropical Australian MPAs further demonstrated that ecological changes continue to develop in MPAs over at least two decades, probably much longer. The continent-scale study additionally showed recently established MPAs to be consistently located at sites with low resource value relative to adjacent fished reference areas. This outcome was presumably generated by sociopolitical pressures and planning processes that aim to systematically avoid locations with valuable resources, potentially compromising biodiversity conservation goals. Locations that were formerly highly fished are needed within MPA networks if the networks are to achieve conservation aims associated with (1) safeguarding all regional habitat types, (2) protecting threatened habitats and species, and (3) providing appropriate reference benchmarks for assessing impacts of fishing. Because of long time lags, the ubiquity of fishing impacts, and the relatively recent establishment of MPAs, the full impact of fishing on coastal reefs has yet to be empirically assessed. PMID- 20014572 TI - Declines in predatory fish promote bloom-forming macroalgae. AB - In the Baltic Sea, increased dominance of ephemeral and bloom-forming algae is presently attributed to increased nutrient loads. Simultaneously, coastal predatory fish are in strong decline. Using field data from nine areas covering a 700-km coastline, we examined whether formation of macroalgal blooms could be linked to the composition of the fish community. We then tested whether predator or nutrient availability could explain the field patterns in two small-scale field experiments, by comparing joint effects on algal net production from nutrient enrichment with agricultural fertilizer and exclusion of larger predatory fish with cages. We also manipulated the presence of invertebrate grazers. The abundance of piscivorous fish had a strong negative correlation with the large-scale distribution of bloom-forming macroalgae. Areas with depleted top predator communities displayed massive increases in their prey, small-bodied fish, and high covers of ephemeral algae. Combining the results from the two experiments showed that excluding larger piscivorous fish: (1) increased the abundance of small-bodied predatory fish; (2) changed the size distribution of the dominating grazers, decreasing the smaller gastropod scrapers; and (3) increased the net production of ephemeral macroalgae. Effects of removing top predators and nutrient enrichment were similar and additive, together increasing the abundance of ephemeral algae many times. Predator effects depended on invertebrate grazers; in the absence of invertebrates there were no significant effects of predator exclusion on algal production. Our results provide strong support for regional declines of larger predatory fish in the Baltic Sea promoting algal production by decreasing invertebrate grazer control. This highlights the importance of trophic interactions for ecosystem responses to eutrophication. The view emerges that to achieve management goals for water quality we need to consider the interplay between top-down and bottom-up processes in future ecosystem management of marine resources. PMID- 20014573 TI - Oceanic, riverine, and genetic influences on spring chinook salmon migration timing. AB - Migrating salmonids often return to their spawning habitats in overlapping timing patterns of multiple stocks (populations) collectively called a run that varies in its genetic makeup across and within years. Managers, tasked with developing harvest strategies on these runs, may have preseason estimates of total run size but little information on run timing. Without both it is difficult to assess a run's status in real time. Consequently, to avoid overharvest, managers tend to control the timing of harvest. However, this strategy may inadvertently affect the component stocks disproportionately and therefore the run's diversity. Thus, accurate estimates of run timing are needed to improve management. We developed a model that includes genetic and environmental factors to predict the mean run timing of chinook salmon (Oncorhynchus tshawytscha) at Bonneville Dam on the Columbia River, Oregon, USA. The model predicted mean runtiming (P < 0.00001, r2 = 0.78) by characterizing genetic run timing components from the arrival timing of precocious males returning one year prior to the remainder of the adults and environmental influences of oceanic and riverine flows that impede or advance the run timing. Variations in the relative abundances of the populations in the run explain 62% of the interannual variation in mean run timing while the oceanic and riverine factors combined account for 15.5%. We suggest that when genetic run timing characteristics are preserved in species with multiple maturation strategies the information can be used to improve run time predictions and maintain genetic diversity of harvested species. PMID- 20014574 TI - A fish of many scales: extrapolating sublethal pesticide exposures to the productivity of wild salmon populations. AB - For more than a decade, numerous pesticides have been detected in river systems of the western United States that support anadromous species of Pacific salmon and steelhead. Over the same interval, several declining wild salmon populations have been listed as either threatened or endangered under the U.S. Endangered Species Act (ESA). Because pesticides occur in surface waters that provide critical habitat for ESA-listed stocks, they are an ongoing concern for salmon conservation and recovery throughout California and the Pacific Northwest. Because pesticide exposures are typically sublethal, a key question is whether toxicological effects at (or below) the scale of the individual animal ultimately reduce the productivity and recovery potential of wild populations. In this study we evaluate how the sublethal impacts of pesticides on physiology and behavior can reduce the somatic growth of juvenile chinook salmon (Oncorhynchus tshawytscha) and, by extension, subsequent size-dependent survival when animals migrate to the ocean and overwinter in their first year. Our analyses focused on the organophosphate and carbamate classes of insecticides. These neurotoxic chemicals have been widely detected in aquatic environments. They inhibit acetylcholinesterase, an enzyme in the salmon nervous system that regulates neurotransmitter-mediated signaling at synapses. Based on empirical data, we developed a model that explicitly links sublethal reductions in acetylcholinesterase activity to reductions in feeding behavior, food ration, growth, and size at migration. Individual size was then used to estimate size dependent survival during migration and transition to the sea. Individual survival estimates were then integrated into a life-history population projection matrix and used to calculate population productivity and growth rate. Our results indicate that short-term (i.e., four-day) exposures that are representative of seasonal pesticide use may be sufficient to reduce the growth and size at ocean entry of juvenile chinook. The consequent reduction in individual survival over successive years reduces the intrinsic productivity (lambda) of a modeled ocean type chinook population. Overall, we show that exposures to common pesticides may place important constraints on the recovery of ESA-listed salmon species, and that simple models can be used to extrapolate toxicological impacts across several scales of biological complexity. PMID- 20014575 TI - Identifying and prioritizing ungulate migration routes for landscape-level conservation. AB - As habitat loss and fragmentation increase across ungulate ranges, identifying and prioritizing migration routes for conservation has taken on new urgency. Here we present a general framework using the Brownian bridge movement model (BBMM) that: (1) provides a probabilistic estimate of the migration routes of a sampled population, (2) distinguishes between route segments that function as stopover sites vs. those used primarily as movement corridors, and (3) prioritizes routes for conservation based upon the proportion of the sampled population that uses them. We applied this approach to a migratory mule deer (Odocoileus hemionus) population in a pristine area of southwest Wyoming, USA, where 2000 gas wells and 1609 km of pipelines and roads have been proposed for development. Our analysis clearly delineated where migration routes occurred relative to proposed development and provided guidance for on-the-ground conservation efforts. Mule deer migration routes were characterized by a series of stopover sites where deer spent most of their time, connected by movement corridors through which deer moved quickly. Our findings suggest management strategies that differentiate between stopover sites and movement corridors may be warranted. Because some migration routes were used by more mule deer than others, proportional level of use may provide a reasonable metric by which routes can be prioritized for conservation. The methods we outline should be applicable to a wide range of species that inhabit regions where migration routes are threatened or poorly understood. PMID- 20014576 TI - Predicting avian distributions to evaluate spatiotemporal overlap with locust control operations in eastern Australia. AB - Locusts and grasshoppers cause considerable economic damage to agriculture worldwide. The Australian Plague Locust Commission uses multiple pesticides to control locusts in eastern Australia. Avian exposure to agricultural pesticides is of conservation concern, especially in the case of rare and threatened species. The aim of this study was to evaluate the probability of pesticide exposure of native avian species during operational locust control based on knowledge of species occurrence in areas and times of application. Using presence absence data provided by the Birds Australia Atlas for 1998 to 2002, we developed a series of generalized linear models to predict avian occurrences on a monthly basis in 0.5 degrees grid cells for 280 species over 2 million km2 in eastern Australia. We constructed species-specific models relating occupancy patterns to survey date and location, rainfall, and derived habitat preference. Model complexity depended on the number of observations available. Model output was the probability of occurrence for each species at times and locations of past locust control operations within the 5-year study period. Given the high spatiotemporal variability of locust control events, the variability in predicted bird species presence was high, with 108 of the total 280 species being included at least once in the top 20 predicted species for individual space-time events. The models were evaluated using field surveys collected between 2000 and 2005, at sites with and without locust outbreaks. Model strength varied among species. Some species were under- or over-predicted as times and locations of interest typically did not correspond to those in the prediction data set and certain species were likely attracted to locusts as a food source. Field surveys demonstrated the utility of the spatially explicit species lists derived from the models but also identified the presence of a number of previously unanticipated species. These results also emphasize the need for special consideration of rare and threatened species that are poorly predicted by presence-absence models. This modeling exercise was a useful a priori approach in species risk assessments to identify species present at times and locations of locust control applications, and to discover gaps in our knowledge and need for further focused data collection. PMID- 20014577 TI - Extrapolating population size from the occupancy-abundance relationship and the scaling pattern of occupancy. AB - The estimation of species abundances at regional scales requires a cost-efficient method that can be applied to existing broadscale data. We compared the performance of eight models for estimating species abundance and community structure from presence-absence maps of the southern African avifauna. Six models were based on the intraspecific occupancy-abundance relationship (OAR); the other two on the scaling pattern of species occupancy (SPO), which quantifies the decline in species range size when measured across progressively finer scales. The performance of these models was examined using five tests: the first three compared the predicted community structure against well-documented macroecological patterns; the final two compared published abundance estimates for rare species and the total regional abundance estimate against predicted abundances. Approximately two billion birds were estimated as occurring in South Africa, Lesotho, and Swaziland. SPO models outperformed the OAR models, due to OAR models assuming environmental homogeneity and yielding scale-dependent estimates. Therefore, OAR models should only be applied across small, homogenous areas. By contrast, SPO models are suitable for data at larger spatial scales because they are based on the scale dependence of species range size and incorporate environmental heterogeneity (assuming fractal habitat structure or performing a Bayesian estimate of occupancy). Therefore, SPO models are recommended for assemblage-scale regional abundance estimation based on spatially explicit presence-absence data. PMID- 20014578 TI - Estimating the effects of detection heterogeneity and overdispersion on trends estimated from avian point counts. AB - Point counts are a common method for sampling avian distribution and abundance. Although methods for estimating detection probabilities are available, many analyses use raw counts and do not correct for detectability. We use a removal model of detection within an N-mixture approach to estimate abundance trends corrected for imperfect detection. We compare the corrected trend estimates to those estimated from raw counts for 16 species using 15 years of monitoring data on three national forests in the western Great Lakes, USA. We also tested the effects of overdispersion by modeling both counts and removal mixtures under three statistical distributions: Poisson, zero-inflated Poisson, and negative binomial. For most species, the removal model produced estimates of detection probability that conformed to expectations. For many species, but not all, estimates of trends were similar regardless of statistical distribution or method of analysis. Within a given combination of likelihood (counts vs. mixtures) and statistical distribution, trends usually differed by both stand type and national forest, with species showing declines in some stand types and increases in others. For three species, Brown Creeper, Yellow-rumped Warbler, and Black throated Green Warbler, temporal patterns in detectability resulted in substantial differences in estimated trends under the removal mixtures compared to the analysis of raw counts. Overall, we found that the zero-inflated Poisson was the best distribution for our data, although the Poisson or negative binomial performed better for a few species. The similarity in estimated trends that we observed among counts and removal mixtures was probably a result of both experimental design and sampling effort. First, the study was originally designed to avoid confounding observer effects with habitats or time. Second, our time series is relatively long and our sample sizes within years are large. PMID- 20014579 TI - Uncertainty analysis of least-cost modeling for designing wildlife linkages. AB - Least-cost models for focal species are widely used to design wildlife corridors. To evaluate the least-cost modeling approach used to develop 15 linkage designs in southern California, USA, we assessed robustness of the largest and least constrained linkage. Species experts parameterized models for eight species with weights for four habitat factors (land cover, topographic position, elevation, road density) and resistance values for each class within a factor (e.g., each class of land cover). Each model produced a proposed corridor for that species. We examined the extent to which uncertainty in factor weights and class resistance values affected two key conservation-relevant outputs, namely, the location and modeled resistance to movement of each proposed corridor. To do so, we compared the proposed corridor to 13 alternative corridors created with parameter sets that spanned the plausible ranges of biological uncertainty in these parameters. Models for five species were highly robust (mean overlap 88%, little or no increase in resistance). Although the proposed corridors for the other three focal species overlapped as little as 0% (mean 58%) of the alternative corridors, resistance in the proposed corridors for these three species was rarely higher than resistance in the alternative corridors (mean difference was 0.025 on a scale of 1 10; worst difference was 0.39). As long as the model had the correct rank order of resistance values and factor weights, our results suggest that the predicted corridor is robust to uncertainty. The three carnivore focal species, alone or in combination, were not effective umbrellas for the other focal species. The carnivore corridors failed to overlap the predicted corridors of most other focal species and provided relatively high resistance for the other focal species (mean increase of 2.7 resistance units). Least-cost modelers should conduct uncertainty analysis so that decision-makers can appreciate the potential impact of model uncertainty on conservation decisions. Our approach to uncertainty analysis (which can be called a worst-case scenario approach) is appropriate for complex models in which distribution of the input parameters cannot be specified. PMID- 20014580 TI - Landscape characteristics of a stream and wetland mitigation banking program. AB - In the United States, stream restoration is an increasing part of environmental and land management programs, particularly under the auspices of compensatory mitigation regulations. Markets and regulations surrounding stream mitigation are beginning to mirror those of the well-established wetland mitigation industry. Recent studies have shown that wetland mitigation programs commonly shift wetlands across space from urban to rural areas, thereby changing the functional characteristics and benefits of wetlands in the landscape. However, it is not yet known if stream mitigation mirrors this behavior, and if so, what effects this may have on landscape-scale ecological and hydrological processes. This project addresses three primary research questions. (1) What are the spatial relationships between stream and wetland impact and compensation sites as a result of regulations requiring stream and wetland mitigation in the State of North Carolina? (2) How do stream impacts come about due to the actions of different types of developers, and how do the characteristics of impacts sites compare with compensation sites? (3) To what extent does stream compensation relocate high-quality streams within the river network, and how does this affect localized (intrawatershed) loss or gain of aquatic resources? Using geospatial data collected from the North Carolina Division of Water Quality and the Army Corps of Engineers' Wilmington District, we analyzed the behavior of the North Carolina Ecosystem Enhancement Program in providing stream and wetland mitigation for the State of North Carolina. Our results suggest that this program provides mitigation (1) in different ways for different types of permittees; (2) at great distances (both Euclidean and within the stream network) from original impacts; (3) in significantly different places than impacts within watersheds; and (4) in many cases, in different watersheds from original impacts. Our analysis also reveals problems with regulator data collection, storage, and quality control. These results have significant implications given new federal requirements for ecological consistency within mitigation programs. Our results also indicate some of the landscape-scale implications of using market-based approaches to ecological restoration in general. PMID- 20014581 TI - Trajectories of vegetation-based indicators used to assess wetland restoration progress. AB - Temporal trends in attributes of restored ecosystems have been described conceptually as restoration trajectories. Measures describing the maturity or ecological integrity of a restoration site are often assumed to follow monotonically increasing trajectories over time and to eventually reach an asymptote representative of a reference ecosystem. This assumption of simple, predictable restoration trajectories underpins federal and state policies in the United States that mandate wetland restoration as compensation for wetlands damaged during development. We evaluated the validity of this assumption by tracking changes in 11 indicators of floristic integrity, often used to determine legal compliance, in 29 mitigation wetlands. Each indicator was expressed as a percentile relative to the distribution of that indicator among > 100 naturally occurring reference wetlands. Nonlinear regression was used to fit two alternative restoration trajectories to data from each site: an asymptotic (negative exponential) increase in the indicator over time and a peaked (double exponential) relationship. Depending on the particular indicator, between 48% and 76% of sites displayed trends that were at least moderately well described (R2 > 0.5) by one of the two models. Floristic indicators based on species richness, including native richness, number of native genera, and the floristic quality index, rapidly increased to asymptotes exceeding levels in a majority of reference wetlands. In contrast, indicators based on species composition, including mean coefficient of conservatism and relative importance of perennial species, increased very slowly. Thus, some indicators of restoration progress followed increasing trajectories and achieved or surpassed levels equivalent to high-quality reference sites within five years, whereas others appeared destined to either not reach equivalency or to take much longer than mitigation wetlands are typically monitored. Finally, some indicators of restoration progress, such as relative importance of native species, often increased over the first five to 10 years and then declined, which would result in a misleading assessment of progress if based on typical time scales of monitoring. Therefore, the assumption of simple, rapid, and predictable restoration trajectories that underlies wetland mitigation policy is unrealistic. PMID- 20014582 TI - Relative influence of landscape vs. local factors on plant community assembly in restored wetlands. AB - Ecological restoration often involves only the manipulation of abiotic factors at the local scale. However, processes external to a restoration site determine the range of local conditions within the site, constraining the level of restoration progress that can be achieved by on-site manipulations. We examined the relationship of landscape and local explanatory variables to plant species composition in 28 restored wetlands in Illinois, USA. Using constrained ordination combined with variation partitioning, we determined the independent and joint effects of three spatially hierarchical sets of variables: (1) macroscale landscape features reflecting site setting within regional landscapes, (2) mesoscale landscape features reflecting nearby propagule sources and buffers from disturbances, and (3) local environmental factors. Because the relative influence of landscape- vs. local-scale factors on restoration success may depend on particular restoration goals, we repeated the analyses using three multivariate plant community responses that represented three frequently stated goals: (1) replicating species composition, (2) restoring a particular wetland community type, and (3) constructing sites with high value for plant conservation. Explanatory variables at landscape and local scales had independent and nearly equally strong relationships to plant species composition. In contrast, when species were aggregated based on plant traits, the independent contribution of local predictors was greater than the independent contributions of macroscale or mesoscale landscape predictors, reflecting convergence of plant trait composition in sites with similar local conditions. Local predictors explained a significant amount of variation in plant conservation value among sites, but much of the variation could be explained by large-scale landscape setting, indicating that landscape constraints on local environmental conditions limited the level of floristic conservation value achievable. The appropriate scale at which to focus restoration efforts will vary depending upon restoration objectives. Restoration of particular wetland community types might be successfully achieved through manipulation of local abiotic factors. In contrast, restoration of a particular species assemblage or reconstruction of wetlands with high value for conservation requires consideration of landscape processes and available species pools. PMID- 20014583 TI - Are pine plantations valid tools for restoring Mediterranean forests? An assessment along abiotic and biotic gradients. AB - The ecological impacts of forest plantations are a focus of intense debate, from studies that consider plantations as "biological deserts" to studies showing positive effects on plant diversity and dynamics. This lack of consensus might be influenced by the scarcity of studies that examine how the ecological characteristics of plantations vary along abiotic and biotic gradients. Here we conducted a large-scale assessment of plant regeneration and diversity in plantations of southern Spain. Tree seedling and sapling density, plant species richness, and Shannon's (H') diversity index were analyzed in 442 pine plantation plots covering a wide gradient of climatic conditions, stand density, and distance to natural forests that act as seed sources. Pronounced variation in regeneration and diversity was found in plantation understories along the gradients explored. Low- to mid-altitude plantations showed a diverse and abundant seedling bank dominated by Quercus ilex, whereas high-altitude plantations showed a virtually monospecific seeding bank of Pinus sylvestris. Regeneration was null in plantations with stand densities exceeding 1500 pines/ha. Moderate plantation densities (500-1000 pines/ha) promoted recruitment in comparison to low or null canopy cover, suggesting the existence of facilitative interactions. Quercus ilex recruitment diminished exponentially with distance to the nearest Q. ilex forest. Richness and H' index values showed a hump-shaped distribution along the altitudinal and radiation gradients and decreased monotonically along the stand density gradient. From a management perspective, different strategies will be necessary depending on where a plantation lies along the gradients explored. Active management will be required in high-density plantations with arrested succession and low diversity. Thinning could redirect plantations toward more natural densities where facilitation predominates. Passive management might be recommended for low- to moderate density plantations with active successional dynamics (e.g., toward oak or pine oak forests at low to mid altitudes). Enrichment planting will be required to overcome seed limitation, especially in plantations far from natural forests. We conclude that plantations should be perceived as dynamic systems where successional trajectories and diversity levels are determined by abiotic constraints, complex balances of competitive and facilitative interactions, the spatial configuration of native seed sources, and species life-history traits. PMID- 20014584 TI - Alpha and beta diversity of plants and animals along a tropical land-use gradient. AB - Assessing the overall biological diversity of tropical rain forests is a seemingly insurmountable task for ecologists. Therefore, researchers frequently sample selected taxa that they believe reflect general biodiversity patterns. Usually, these studies focus on the congruence of alpha diversity (the number of species found per sampling unit) between taxa rather than on beta diversity (turnover of species assemblages between sampling units). Such approaches ignore the potential role of habitat heterogeneity that, depending on the taxonomic group considered, can greatly enhance beta diversity at local and landscape scales. We compared alpha and beta diversity of four plant groups (trees, lianas, terrestrial herbs, epiphytic liverworts) and eight animal groups (birds, butterflies, lower canopy ants, lower canopy beetles, dung beetles, bees, wasps, and the parasitoids of the latter two) at 15 sites in Sulawesi, Indonesia, that represented natural rain forest and three types of cacao agroforests differing in management intensity. In total, we recorded 863 species. Patterns of species richness per study site varied strongly between taxonomic groups. Only 13-17% of the variance in species richness of one taxonomic group could be predicted from the species richness of another, and on average 12-18% of the variance of beta diversity of a given group was predicted by that in other groups, although some taxon pairs had higher values (up to 76% for wasps and their parasitoids). The degree of congruence of patterns of alpha diversity was not influenced by sampling completeness, whereas the indicator value for beta diversity improved when using a similarity index that accounts for incomplete sampling. The indication potential of alpha diversity for beta diversity and vice versa was limited within taxa (7-20%) and virtually nil between them (0-4%). We conclude that different taxa can have largely independent patterns of alpha diversity and that patterns of beta diversity can be more congruent. Thus, conservation plans on a landscape scale need to put more emphasis on the high heterogeneity of agroforests and the overarching role of beta diversity shaping overall diversity patterns. PMID- 20014585 TI - Analysis of the predator community of a subterranean herbivorous insect based on polymerase chain reaction. AB - The identity and impact of trophic linkages within subterranean arthropod communities are challenging to establish, a fact that hinders the development of conservation biological control programs of subterranean herbivores. Diabrotica virgifera (the western corn rootworm) is a severe agricultural pest that lives subterraneously during its pre-imaginal stages and succumbs to high levels of pre imaginal mortality from unknown agents. The guts of 1500 field-collected arthropod predators were analyzed for D. virgifera-specific DNA sequences using quantitative polymerase chain reaction (qPCR). These gut analyses were used to generate relative and taxon-specific prey consumption indices for the major predator taxa and to determine relative consumption levels during D. virgifera egg and larval stages by predator feeding guilds. Laboratory feeding assays were used to determine the meal size consumed during 5 min and digestion rates of D. virgifera DNA of four predators abundant in D. virgifera-infested cornfields. More than 17 taxa consumed D. virgifera in the field. Harvestmen and small rove beetles were the most abundant predators captured, and the most frequent predators within the community to consume D. virgifera. The largest proportions of individual species' populations testing positive for D. virgifera DNA were found in ground beetles (Scarites quadriceps and Poecilus chalcites) and spiders, wolf spiders, and predaceous mites. Because of the longer duration of the egg stage, significantly more predators consumed D. virgifera eggs than larvae, but a similar proportion of the predator community fed on eggs and larvae. Predators with sucking mouthparts had a higher consumption index than chewing predators. Laboratory assays confirmed that sucking predators consume more D. virgifera DNA during 5 min than the chewing predators, and all four predators digested this DNA at a similar rate. This research substantiates that a diverse community of soil dwelling and subterranean predators contribute to the high level of mortality incurred by D. virgifera in cornfields (approximately 99% pre-adult mortality). Moreover, qPCR is a useful tool for describing trophic relationships within subterranean food webs, a crucial step in determining the relative contributions of a diverse predator community to the population dynamics of an herbivorous arthropod. PMID- 20014586 TI - The fate of nitrogen in grain cropping systems: a meta-analysis of 15N field experiments. AB - Intensively managed grain farms are saturated with large inputs of nitrogen (N) fertilizer, leading to N losses and environmental degradation. Despite decades of research directed toward reducing N losses from agroecosystems, progress has been minimal, and the currently promoted best management practices are not necessarily the most effective. We investigated the fate of N additions to temperate grain agroecosystems using a meta-analysis of 217 field-scale studies that followed the stable isotope 15N in crops and soil. We compared management practices that alter inorganic fertilizer additions, such as application timing or reduced N fertilizer rates, to practices that re-couple the biogeochemical cycles of carbon (C) and N, such as organic N sources and diversified crop rotations, and analyzed the following response variables: 15N recovery in crops, total recovery of 15N in crops and soil, and crop yield. More of the literature (94%) emphasized crop recovery of 15N than total 15N recovery in crops and soil (58%), though total recovery is a more ecologically appropriate indicator for assessing N losses. Findings show wide differences in the ability of management practices to improve N use efficiency. Practices that aimed to increase crop uptake of commercial fertilizer had a lower impact on total 15N recovery (3-21% increase) than practices that re-coupled C and N cycling (30-42% increase). A majority of studies (66%) were only one growing season long, which poses a particular problem when organic N sources are used because crops recover N from these sources over several years. These short-term studies neglect significant ecological processes that occur over longer time scales. Field-scale mass balance calculations using the 15N data set show that, on average, 43 kg N x ha(-1) x yr(-1) was unaccounted for at the end of one growing season out of 114 kg N x ha(-1) x yr(-1), representing approximately 38% of the total 15N applied. This comprehensive assessment of stable-isotope research on agroecosystem N management can inform the development of policies to mitigate nonpoint source pollution. Nitrogen management practices that most effectively increase N retention are not currently being promoted and are rare on the landscape in the United States. PMID- 20014587 TI - Prairie restoration and carbon sequestration: difficulties quantifying C sources and sinks using a biometric approach. AB - We investigated carbon cycling and ecosystem characteristics among two prairie restoration treatments established in 1987 and adjacent cropland, all part of the Conservation Reserve Program in southwestern Wisconsin, USA. We hypothesized that different plant functional groups (cool-season C3 vs. warm-season C4 grasses) between the two prairie restoration treatments would lead to differences in soil and vegetation characteristics and amount of sequestered carbon, compared to the crop system. We found significant (P < 0.05) differences between the two prairie restoration treatments in soil CO2 respiration and above- and belowground productivity, but no significant differences in long-term (approximately 16-year) carbon sequestration. We used a biometric approach aggregating short-term observations of above- and belowground productivity and CO2 respiration to estimate total net primary production (NPP) and net ecosystem production (NEP) using varied methods suggested in the literature. Net ecosystem production is important because it represents the ecosystem carbon sequestration, which is of interest to land managers and policymakers seeking or regulating credits for ecosystem carbon storage. Such a biometric approach would be attractive because it might offer the ability to rapidly assess the carbon source/sink status of an ecosystem. We concluded that large uncertainties in (1) estimating aboveground NPP, (2) determining belowground NPP, and (3) partitioning soil respiration into microbial and plant components strongly affect the magnitude, and even the sign, of NEP estimates made from aggregating its components. A comparison of these estimates across treatments could not distinguish differences in NEP, nor the absolute sign of the overall carbon balance. Longer-term quantification of carbon stocks in the soil, periodically linked to measurements of individual processes, may offer a more reliable measure of the carbon balance in grassland systems, suitable for assigning credits. PMID- 20014588 TI - Plant species composition and biofuel yields of conservation grasslands. AB - Marginal croplands, such as those in the Conservation Reserve Program (CRP), have been suggested as a source of biomass for biofuel production. However, little is known about the composition of plant species on these conservation grasslands or their potential for ethanol production. Our objective was to assess the potential of CRP and other conservation grasslands for biofuel production, describing the relationships of plant species richness and tall native C4 prairie grass abundance with plant chemical composition and the resulting potential ethanol yield. We determined plant species composition and diversity at multiple scales with the modified Whittaker plot technique, aboveground biomass, plant chemical composition, and potential ethanol yield at 34 sites across the major ecological regions of the northeastern USA. Conservation grasslands with higher numbers of plant species had lower biomass yields and a lower ethanol yield per unit biomass compared with sites with fewer species. Thus, biofuel yield per unit land area decreased by 77% as plant species richness increased from 3 to 12.8 species per m2. We found that, as tall native C4 prairie grass abundance increased from 1.7% to 81.6%, the number of plant species decreased and aboveground biomass per unit land area and ethanol yield per unit biomass increased resulting in a 500% increased biofuel yield per unit land area. Plant species richness and composition are key determinants of biomass and ethanol yields from conservation grasslands and have implications for low-input high-diversity systems. Designing systems to include a large proportion of species with undesirable fermentation characteristics could reduce ethanol yields. PMID- 20014589 TI - The relative importance of disturbance and exotic-plant abundance in California coastal sage scrub. AB - Many ecosystems of conservation concern require some level of disturbance to sustain their species composition and ecological function. However, inappropriate disturbance regimes could favor invasion or expansion of exotic species. In southern California coastal sage scrub (CSS) fire is a natural disturbance, but because of human influence, frequencies may now be unnaturally high. Other anthropogenic disturbances such as grazing also occur in reserve areas. Managers charged with imposing or tolerating fire or other disturbance within their reserves are concerned that habitat quality may be degraded by an increasing abundance of exotic plants. We used vegetation monitoring data from Camp Pendleton, California, USA, to assess the correlation between past disturbances (frequent fire, agriculture, or grazing and mechanical disturbances) and current exotic species abundance in CSS. We found that disturbance history was only modestly related to exotic abundance overall, but fire frequency showed the strongest association. We also examined whether cover and richness of various native plant life forms (woody species, perennial herbs, and annual herbs) were more strongly influenced by disturbance history or by exotic-plant abundance. Native plant responses varied among life forms, but woody species and annual herbs were generally more strongly and negatively associated with exotic abundance than with disturbance. Effective CSS conservation will require developing means to curb the negative impacts of exotic plants, which may abound with or without severe or recent disturbance. Additionally, more focus should be given to understory herbs showing sensitivity to invasion. Though understudied, native herbs comprise the greatest portion of plant diversity in CSS and are critical to preservation of the community as a whole. PMID- 20014590 TI - A global meta-analysis of soil exchangeable cations, pH, carbon, and nitrogen with afforestation. AB - Afforestation, the conversion of non-forested lands to forest plantations, can sequester atmospheric carbon dioxide, but the rapid growth and harvesting of biomass may deplete nutrients and degrade soils if managed improperly. The goal of this study is to evaluate how afforestation affects mineral soil quality, including pH, sodium, exchangeable cations, organic carbon, and nitrogen, and to examine the magnitude of these changes regionally where afforestation rates are high. We also examine potential mechanisms to reduce the impacts of afforestation on soils and to maintain long-term productivity. Across diverse plantation types (153 sites) to a depth of 30 cm of mineral soil, we observed significant decreases in nutrient cations (Ca, K, Mg), increases in sodium (Na), or both with afforestation. Across the data set, afforestation reduced soil concentrations of the macronutrient Ca by 29% on average (P < 0.05). Afforestation by Pinus alone decreased soil K by 23% (P < 0.05). Overall, plantations of all genera also led to a mean 71% increase of soil Na (P < 0.05). Mean pH decreased 0.3 units (P < 0.05) with afforestation. Afforestation caused a 6.7% and 15% (P < 0.05) decrease in soil C and N content respectively, though the effect was driven principally by Pinus plantations (15% and 20% decrease, P < 0.05). Carbon to nitrogen ratios in soils under plantations were 5.7-11.6% higher (P < 0.05). In several regions with high rates of afforestation, cumulative losses of N, Ca, and Mg are likely in the range of tens of millions of metric tons. The decreases indicate that trees take up considerable amounts of nutrients from soils; harvesting this biomass repeatedly could impair long-term soil fertility and productivity in some locations. Based on this study and a review of other literature, we suggest that proper site preparation and sustainable harvest practices, such as avoiding the removal or burning of harvest residue, could minimize the impact of afforestation on soils. These sustainable practices would in turn slow soil compaction, erosion, and organic matter loss, maintaining soil fertility to the greatest extent possible. PMID- 20014591 TI - Not all ski slopes are created equal: disturbance intensity affects ecosystem properties. AB - In mountain regions around the world, downhill ski areas represent a significant source of anthropogenic disturbance while also providing recreation and revenue. Ski-run creation always results in some level of disturbance, but disturbance intensity varies greatly with construction method. Ski runs may be established either by clearing (cutting and removing tall vegetation) or by clearing and then machine-grading (leveling the soil surface with heavy equipment). To quantify how these different intensities of initial disturbance affect ecosystem properties, we extensively surveyed vegetation, soils, and environmental characteristics on cleared ski runs, graded ski runs, and adjacent reference forests across seven large downhill ski resorts in the northern Sierra Nevada, USA. We found that the greater disturbance intensity associated with grading resulted in greater impacts on all ecosystem properties considered, including plant community composition and diversity, soil characteristics relating to processes of nutrient cycling and retention, and measures of erosion potential. We also found that cleared ski runs retained many ecological similarities to reference forests and might even offer some added benefits by possessing greater plant species and functional diversity than either forests or graded runs. Because grading is more damaging to multiple indicators of ecosystem function, clearing rather than grading should be used to create ski slopes wherever practical. PMID- 20014592 TI - Swan-Ganz (pulmonary artery) catheters. PMID- 20014593 TI - Low flows revisited. PMID- 20014594 TI - The efficacy of landiolol for suppressing the hyperdynamic response following laryngoscopy and tracheal intubation: a systematic review. AB - Landiolol is a recently developed, selective short-acting beta1-antagonist. The aim of the study was to evaluate the efficacy of landiolol for suppressing haemodynamic changes induced by laryngoscopy and tracheal intubation (LTI) in Japanese patients. A comprehensive search was undertaken to identify all randomised comparisons of landiolol with placebo that examined effects on haemodynamic responses following LTI. MEDLINE, Cochrane CENTRAL, EMBASE and the Japanese Central Review of Medicine were searched from their date of inception to February 2009. Trials were included in the review if heart rate, systolic blood pressure or mean blood pressure was recorded at three different stages: pre induction, just before intubation and in the post-intubation period. Weighted mean differences and 95% confidence intervals (CI) were calculated for changes in haemodynamic variables between treatment and placebo groups. Seven randomised controlled trials involving 325 patients were included in the study. Of these, five trials that used the same continuous infusion regimen for landiolol (0.125 mg/kg/minute for one minute followed by 0.04 mg/kg/minute) showed efficacy in attenuation of heart rate and blood pressure following LTI (heart rate weighted mean difference: -21.18 bpm, 95% CI -18.59 to -14.20; systolic blood pressure weighted mean difference: -23.03 mmHg, 95% CI -43.59 to -2.47; mean blood pressure weighted mean difference: -16.26 mmHg, 95% CI -23.96 to -8.55). The other two studies used bolus administration of landiolol (0.1 to 0.3 mg/kg), but it was difficult to evaluate the efficacy because of the limited amount of data. Landiolol administration at 0.125 mg/kg/minute for one minute followed by 0.04 mg/kg/minute effectively suppresses the increases in heart rate and blood pressure following LTI. For a bolus regimen of landiolol, further studies are required to determine the efficacy and the optimal dose and timing for suppression of haemodynamic responses following LTI. PMID- 20014595 TI - The rise of simulation in technical skills teaching and the implications for training novices in anaesthesia. AB - Changes in work practices have led to a decline in the opportunities for anaesthetic trainees to learn technical procedures in supervised practice. Efforts to mitigate medical error and other changes have coincided with the development of alternative training methods so that it is increasingly difficult to justify the traditional model of teaching technical procedures. The range of simulators available for training in technical procedures in anaesthesia continues to expand. While simulation has been widely adopted in anaesthesia for crisis management training, there is little documented evidence of its use for technical skills training. The theoretical basis for the use of simulation to aid the acquisition of psychomotor skills and the development of expertise is now well established. In addition, practical frameworks that allow this theory to be applied in a systematic fashion have been developed and successfully used in other specialties. Using the available simulation equipment and educational tools, trainees can be prepared to begin supervised practice having demonstrated adequate procedural knowledge and expertise in simulation. With the use of simulated patients there is also the opportunity to integrate non-technical skills as well where appropriate. This review summarises the justification for the use of simulation in technical skills training in anaesthesia and the educational theory that supports its use, and outlines one of the available frameworks that can be used to aid its application. PMID- 20014596 TI - Derivation and prospective testing of a two-step sevoflurane-O2-N2O low fresh gas flow sequence. AB - Simple vaporiser setting (F(D)) and fresh gas flow (FGF) sequences make the practice of low-flow anaesthesia not only possible but also easy to achieve. We sought to derive a sevoflurane F(D) sequence that maintains the end-expired sevoflurane concentration (F(A)sevo) at 1.3% using the fewest possible number of F(D) adjustments with a previously described O2-N2O FGF sequence that allows early FGF reduction to 0.7 l min(-1). In 18 ASA physical status I to IH patients, F(D) was determined to maintain F(A)sevo at 1.3% with 2 l min(-1) O2 and 4 l min( 1) N2O FGF for three minutes, and with 0.3 and 0.4 l min(-1) thereafter. Using the same FGF sequence, the F(D) schedule that approached the 1.3% F(A)sevo pattern with the fewest possible adjustments was prospectively tested in another 18 patients. The following F(D) sequence approximated the F(D) course well: 2% from zero to three minutes, 2.6% from three to 15 minutes and 2.0% after 15 minutes. When prospectively tested, median (25th; 75th percentile) performance error was 0.8 (-2.9; 5.9)%, absolute performance error 6.7 (3.3; 10.6)%, divergence 18.2 (-5.6; 27.4)%.h(-1) and wobble 4.4 (1.7; 8.1) %. In one patient, FGF had to be temporarily increased for four minutes. One O2/N2O rotameter FGF setting change from 6 to 0.7 l min(-1) at three minutes and two sevoflurane F(D) changes at three and 15 minutes maintained predictable anaesthetic gas concentrations during the first 45 minutes in all but one patient in our study. PMID- 20014597 TI - Continuous intra-articular infusion of ropivacaine after unilateral total knee arthroplasty. AB - Intra-articular infusion of local anaesthetic after joint arthroplasty is attractive in that it is simple and will not cause motor block. However the efficacy of the technique has yet to be established. We enrolled 66 patients scheduled for unilateral total knee arthroplasty under general anaesthesia and single-shot femoral and sciatic nerve blocks. All patients had an intra-articular Painbuster device sited at the end of the procedure. Patients were then randomised to control or one of two treatment arms - low-dose and high-dose ropivacaine. In the control group the balloon was filled with saline, in the low dose group with 0.2% ropivacaine and in the high-dose group 0.375% ropivacaine. The catheters were infused continuously for 48 hours and then removed. Patients were followed up daily for three days to determine pain scores, opioid consumption and subjective assessment of the analgesic efficacy of the catheter Data were analysed for 30 controls and 31 in the treatment arms. Both groups were similar There were no significant differences between the control and treatment groups at all time points after adjustment for age. Patients in the high-dose group had higher pain scores and higher opioid consumption than the control groups from 24 to 48 hours. There were two cases of infection, both in the treatment groups. No positive benefit of intra-articular infusion of local anaesthetic after total knee arthroplasty could be identified. On the contrary there may be negative effects in terms of expense, pain and possibly infection risks. PMID- 20014599 TI - Repeated intrathecal administration of ropivacaine causes neurotoxicity in rats. AB - Previous studies suggest that ropivacaine causes the least neurotoxicity among local anaesthetics. Most data derive from a single injection of ropivacaine into the subarachnoid space. The histological changes and behavioural effects of repeated intrathecal administration have yet to be studied. We examined the possible neurotoxicity of multiple doses of intrathecal ropivacaine in rats. Rats received 0.12 ml/kg body weight ropivacaine in normal saline at concentrations of 0.25%, 0.5%, 0.75% and 1.0% at 90-minute intervals via an implanted intrathecal catheter (ID 0.12 mm, OD 0.35 mm) for 48 hours. At L3, the spinal cord and posterior roots were examined by light and electron microscopy. We performed in situ TUNEL assay to evaluate apoptosis in the spinal cord. Sensory threshold to noxious stimulation along with behavioural change were also studied. Both 0.75% and 1.0% ropivacaine induced neuronal injury characterised by infiltration of inflammatory cells, vacuolation of myelin sheaths and axons, abnormal morphology of neurons and apoptosis in the spinal cord, mainly in posterior roots and the adjacent posterior white matter Compared to controls, the percentage of maximum possible effect did not show any significant differences between the rats treated with variable concentrations of ropivacaine or tested with either heat or mechanical stimulation. As expected, the recovery time to normal ambulation was prolonged as the ropivacaine concentration was increased. Ropivacaine can induce neurotoxicity and trigger apoptosis in a dose-dependent manner after repeated intrathecal administration. Although the clinical safety profile of ropivacaine appears favourable compared with other local anaesthetics, it is possible our findings have clinical significance. PMID- 20014598 TI - Comparison of equipotent doses of ropivacaine-fentanyl and bupivacaine-fentanyl in spinal anaesthesia for lower abdominal surgery. AB - The aim of this randomised, double-blind study was to compare equipotent doses of plain ropivacaine and bupivacaine (19.5 mg and 13 mg respectively), both with fentanyl 20 microg, for spinal anaesthesia in lower abdominal surgery. After written informed consent had been obtained, 52 ASA I to II male patients scheduled for lower abdominal surgery were randomly assigned to receive intrathecal plain ropivacaine 19.5 mg with fentanyl 20 microg (group R, n =26) or plain bupivacaine 13 mg with fentanyl 20 microg (group B, n =26) in 3 ml. The level and duration of sensory block, intensity and duration of motor block, time to mobilise and patient satisfaction were recorded. All patients achieved sensory block to T10 or higher The level of sensory block was significantly higher in group B (T4 [T3 to T7] vs T7 [T4 to T9], P <0.05). There was no difference in the onset time of motor block. The duration of motor block (Bromage score >0) was shorter in group R (139+/-39 minutes vs group B 182+/-46 minutes, P <0.05). The duration and intensity of complete motor block (Bromage score=3) were also shorter in group R (90+/-25 minutes vs 130+/-40 minutes, P <0.05). We conclude that plain ropivacaine 19.5 mg plus fentanyl 20 microg is associated with a lower level of sensory block and a shorter duration of motor block when compared to bupivacaine 13 mg plus fentanyl 20 microg for spinal anaesthesia in lower abdominal surgery. PMID- 20014600 TI - The effects of prostaglandin E1 on interleukin-6, pulmonary function and postoperative recovery in oesophagectomised patients. AB - The inflammatory reactions and tissue response after oesophagectomy are leading causes of postoperative morbidity and mortality. We evaluated the effects of intraoperative infusion of prostaglandin E1 (PGE1) on interleukin-6 (IL-6) levels, (A-a) DO2, pulmonary function and complications. This randomised double blind clinical trial study was performed on patients undergoing transthoracic oesophagectomy due to cancer Thirty patients were randomly allocated to two groups: the PGE1 group (infusion of PGE1 20 ng kg(-1) min(-1)) and a placebo group (infusion of normal saline 0.9%). The infusion was started before induction of anaesthesia and continued until the end of the operation. The groups were comparable in basic characteristics and preoperative pulmonary function. Patients in the PGE1 group were discharged significantly earlier from the intensive care unit (72+/-9 vs 83+/-17 hours) and hospital (13+/-4 vs 18+/-8 days) (P=0.04 and 0.03, respectively). The (A-a) DO2 was significantly less in the PGE1 group at 12 and 24 hours after the operation (P=0.001, P=0.003, respectively). Postoperatively, IL-6 levels were significantly higher in the placebo group than in the PGE1 group. There were no differences in the forced expiratory volume in the first second or forced vital capacity. The findings indicate that infusion of PGE1 attenuates the increase in serum levels of IL-6 in patients undergoing esophagectomy and improves the (A-a) DO2. Stays in the intensive care unit and hospital were shorter in the PGE1 group. However, there were no differences in pulmonary complications. PMID- 20014601 TI - Aminoglycoside clearance is a good estimate of creatinine clearance in intensive care unit patients. AB - The aim of this study was to determine whether creatinine clearance can be estimated as well by clearance of gentamicin/tobramycin as by routine, non invasive estimates in the intensive care unit. The volume of distribution and clearance values for gentamicin/tobramycin were obtained using first order kinetics and an estimate of creatinine clearance derived. Seven estimates of renal function (Cockroft-Gault, MDRD4 and MDRD6 equations, two- and 24-hour urine estimates, two equations utilising Cystatin C concentrations) were compared to the gentamicin/tobramycin clearance estimate in 100 intensive care unit patients. The gentamicin clearance estimate was at least as reliable as other estimates. The two-hour was less reliable than the 24-hour urine estimate. The Cockroft Gault appeared to out-perform the MDRD equation estimates. The MDRD4 was not as reliable as the MDRD6 estimate. Cystatin C estimates appeared not as reliable as the gentamicin estimate of renal function. The gentamicin/tobramycin estimate is at least as good as other estimates and it is available sooner than most others. It should be used in all patients who are prescribed gentamicin. The two-hour urine and MDRD4 estimates should not be used in the intensive care unit. PMID- 20014602 TI - Comparison of forehead Max-Fast pulse oximetry sensor with finger sensor at high positive end-expiratory pressure in adult patients with acute respiratory distress syndrome. AB - In the critical care setting it may be difficult to determine an accurate reading of oxygen saturation from digital sensors as a result of poor peripheral perfusion. Limited evidence suggests that forehead sensors may be more accurate in these patients. We prospectively compared the accuracy of a forehead reflectance sensor (Max-Fast) with a conventional digital sensor in patients with acute respiratory distress syndrome during a high positive end-expiratory pressure (PEEP) recruitment manoeuvre (stepwise recruitment manoeuvre). Sixteen patients with early acute respiratory distress syndrome were enrolled to evaluate the blood oxygen saturation during a stepwise recruitment manoeuvre. PEEP was increased from baseline (range 10 to 18) to 40 cmH2O, then decreased to an optimal level determined by individual titration. Forehead and digital oxygen saturation and arterial blood gases were measured simultaneously before, during and after the stepwise recruitment manoeuvre at five time points. Seventy-three samples were included for analysis from 16 patients. The SaO2 values ranged from 73 to 99.6%. The forehead sensor provided measurements that deviated more from arterial measures than the finger sensor (mean absolute deviations 3.4%, 1.1% respectively, P=0.02). The greater variability in forehead measures taken at maximum PEEP was reflected in the unusually large precision estimates of 4.24% associated with these measures. No absolute differences from arterial measures taken at any other time points were significantly different. The finger sensor is as accurate as the forehead sensor in detecting changes in arterial oxygen saturation in adults with acute respiratory distress syndrome and it may be better at levels of high PEEP such as during recruitment manoeuvres. PMID- 20014603 TI - Cricothyroidotomy: comparison of three different techniques on a porcine airway. AB - We compared three different cricothyroidotomy techniques on a cadaveric porcine airway model to determine the most rapid, successful and safe method in emergency situations. In this observational comparative bench-test, surgical or scalpel cricothyroidotomy (ST), Cook Melker Cricothyrotomy Kit (CM) and Portex Cricothyroidotomy Kit (PCK) were evaluated. After a familiarisation program, four operators performed five procedures using each of the techniques. A dissected pig airway was housed in an airway training manikin. Time to successful completion of the procedure, complications and anatomical variations in the pig larynx were recorded. A patent airway was established in 20 of 20 (100%) attempts with CM technique, 11 of 20 (55%) with ST and six of 20 (30%) with PCK technique. The CM technique was associated with a higher success rate (P <0.001). There was no significant difference (P=0.4) in the median time (interquartile range) taken to achieve a patent airway; CM 50 seconds (40 to 57), ST 47 seconds (41 to 55) and PCK 63 seconds (41 to 150). There was a higher rate of posterior tracheal wall injury with the PCK technique (n=11) and ST technique (n=5). No complications were observed with CM technique. There were no significant anatomical variations in the pig larynxes. We conclude that the CM technique is safe, rapid and has a significantly higher success rate in achieving an artificial airway in this airway model. Overall, all the participants in our study preferred to use this kit in an emergency situation. PMID- 20014604 TI - Incidence of contrast-induced nephropathy in intensive care patients undergoing computerised tomography and prevalence of risk factors. AB - Computerised tomography (CT) with contrast is frequently used in intensive care. Contrast-induced nephropathy (CIN) is an important complication largely studied in stable cardiology patients and can lead to acute renal failure. The aim of this study was to determine the incidence of CIN in an intensive care unit (ICU) setting and describe the prevalence of associated risk factors. We performed a retrospective analysis by review of electronic laboratory database and manual chart review of all patients in two tertiary intensive care units in Newcastle, New South Wales who underwent CT with intravenous contrast during their ICU stay in 2006. CIN was defined as an absolute increment in serum creatinine of 44.2 micromol/l or a relative increment of 25% from baseline at 48 to 72 hours following intravenous contrast. Patients' demographic, biochemical and contrast media data, physiological parameters, fluid and drug administrations and previously described as well as ICU specific risk factors were analysed. We compared CIN positive and CIN negative patients to identify risk factors associated with CIN. In total, 2043 patients were admitted to ICU during 2006 and 509 CT studies were performed. One hundred and forty-one of these included administration of intravenous contrast and 139 charts were reviewed. Sixteen out of 139 patients developed CIN (11.5%). More than 70% of patients had two or more risk factors. Age was the only risk factor found to be significantly associated with the development of CIN in a multivariate analysis (P value 0.04, OR 1.041, 95% confidence interval 1.002 to 1.081). Mortality was higher in CIN positive patients (31 vs 13%, P value 0.068). ICU and hospital length of stay was not significantly different in CIN positive and negative patients and persisting renal impairment was not found in CIN positive survivors. Based on this study, we cannot predict who will develop CIN in ICU using the described risk factors. Further prospective studies are needed to evaluate the incidence and outcomes of CIN in an ICU setting. PMID- 20014605 TI - Predictors and outcome associated with an Enterococcus positive isolate during intensive care unit admission. AB - This study reports the incidence, risk factors and mortality associated with a positive Enterococcus spp. isolate during admission to two tertiary intensive care units participating in an antibiotic cycling study. Incidence was low, with only 4.2% of admissions (36/852) at Royal Brisbane and Women's Hospital and 2.8% (31/1104) at Westmead Hospital developing a positive Enterococcus spp. isolate (P=0.087). A positive enterococcal isolate, while not an independent predictor of mortality (odds ratio [OR]=1.6, 95% confidence interval [CI] 0.80 to 3.2, P=0.18), may be a marker of the underlying severity of illness with higher unadjusted in-hospital mortality (26% or 17/66 vs 14% or 250/1855, P=0.007). Independent risk factors for a positive isolate were use of meropenem/imipenem (OR=5.7, 95% CI 2.4 to 14, P <0.001) and cefepime (OR=2.5, 95% CI 1.2 to 5.3, P=0.017) within 48 hours of intensive care unit admission, the presence of a nasogastric tube (OR=4.1, 95% CI 1.3 to 14, P=0.018), renal replacement therapy (OR=2.2, 95% CI 1.0 to 4.7, P=0.046), operative intervention (OR=1.8, 95% CI 1.0 to 3.2, P=0.035) and age (OR=1.2, 95% CI 1.1 to 1.5, P=0.009). None of these factors, except for the need for renal replacement therapy (OR=6.2, 95% CI 1.4 to 27, P=0.015), was associated with increased mortality. Enterococci-directed empiric therapy in the treatment of sepsis remains of unproven value, although this negative finding must be evaluated against other higher powered studies. PMID- 20014606 TI - An assessment of the population variance of the strong ion gap using Monte Carlo simulation. AB - The strong ion gap is a derived parameter used to quantify clinically significant but unmeasured charged species in human plasma. In the general population the strong ion gap has a mean value of approximately 0.0 mEq/l, but at present no reference range exists. This severely limits its clinical application. In order to establish a reference range, a 95% confidence interval around the population mean must be calculated. As each of the variables that make up the strong ion gap have their own unique and different means and standard deviations, simple methods of pooled variance can not be used to calculate this interval. In place of a direct solution, Monte Carlo methodology was employed using known reference standard deviations to construct a large sample of a simulated population. In addition, the partial correlation between the plasma concentrations of sodium and chloride was included in the calculations. Sample sets of 20,000 were simulated, each giving a normally distributed strong ion gap. A 95% confidence interval of 3.9+/-6.4 mEq/l was derived. Depending on the degree of strong ion correlation, minima and maxima for this reference range were calculated as 3.9+/-5.1 mEq/l and 3.9+/-7.6 mEq/l respectively. Reasons for the positive bias in the mean strong ion gap result are examined briefly. PMID- 20014607 TI - A randomised, single-blind, controlled trial of silicone disposable laryngeal masks during anaesthesia in spontaneously breathing adult patients. AB - We hypothesised that two new silicone disposable laryngeal mask airways, Meditech Systems Limited (MSL) and ProAct (PA), would perform similarly to the Classic Laryngeal Mask Airway (cLMA) in spontaneously breathing adult patients. One hundred and twenty consecutive adult patients were prospectively enrolled and randomly allocated to one of three groups (PA, MSL, cLMA). All patients received a standardised anaesthetic and insertion technique. The overall success rates were 97.5% for all three masks, with one failure of laryngeal mask airway insertion in each group. There was no statistical difference in airway sealing pressures between the PA laryngeal mask airway (17.9+/-5.9 cmH2O), MSL laryngeal mask airway (18.5+/-6.9 cmH2O,) and cLMA (17.6+/-5.6 cmH2O) (P=0.816). There was no statistical difference in insertion times, ease of insertion, cuff pressure and cuff volumes. The MSL and PA disposable laryngeal mask airways provided comparable airway sealing pressures to the reusable cLMA. PMID- 20014608 TI - Audit of performance of size 1.5 ProSeal laryngeal mask airways in infants less than six months undergoing inguinal herniotomy. AB - Many anaesthetists have found the size 1.5 classic Laryngeal Mask Airway unsuitable for use in children under 10 kg, whereas recent studies evaluating the ProSeal Laryngeal Mask Airway (PLMA) show high success rates, even during laparoscopic surgery. Our routine practice has been to use tracheal intubation for inguinal herniotomy in children weighing less than 10 kg. Following the introduction of the PLMA to our hospital, we decided to audit our use of the PLMA 1.5 in this group of patients. We included 20 consecutive infants, aged less than six months and weighing 5 to 10 kg. We recorded patient, anaesthetic and insertion details, device performance data and complications. No aspect of anaesthetic practice was changed by involvement in this audit. The PLMA was inserted successfully at the first attempt in 85% (17/20) of infants. Overall successful insertion occurred in 90% (18/20) and satisfactory airway maintenance was provided for the duration of anaesthesia in 90% (18/20). The mean leak pressure was 24 cmH2O (range 15 to 30 cmH2O). We found the 1.5 PLMA provided a satisfactory airway in 90% of infants. This report adds to the evidence that the PLMA 1.5 can provide a satisfactory alternative to intubation in selected infants. PMID- 20014609 TI - A failure of M-Entropy to correctly detect burst suppression leading to sevoflurane overdosage. AB - Electroencephalogram depth of anaesthesia monitors are increasingly being used, with the aim of reducing awareness during anaesthesia. Most literature concentrates on the ability of these monitors to predict when a patient is likely to be aware. This case report highlights the opposite problem, where the monitor (M-Entropy) indicated an awake state but the patient was in fact deeply anaesthetised. If the anaesthetist is unable to interpret the raw electroencephalogram and understand the limitations of the monitor being used, excessive doses of anaesthetic may be given with potentially serious consequences. PMID- 20014610 TI - Thyroid storm presenting as intra-abdominal sepsis with multi-organ failure requiring intensive care. AB - Thyroid storm is a rare but life-threatening condition manifesting with several clinical presentations. Atypical thyroid storm should be part of the differential diagnosis in patients with multiple organ dysfunction of unknown aetiology. In this case report, delayed recognition of thyroid storm in a young female who presented with acute abdomen increased the risk of poor outcome. Prompt initiation of anti-thyroid therapy once the diagnosis of thyroid storm was established, combined with adequate vital organ support using a goal-directed therapy protocol in the intensive care unit resulted in a good outcome. PMID- 20014611 TI - Regional anaesthesia and analgesia on the front line. AB - Deployment to a combat zone with the military poses many challenges to the anaesthetist. One of these challenges is the safe, rapid and comfortable initial wound management and repatriation of wounded combat soldiers to their home country or tertiary treatment facility for definitive care and rehabilitation. The current conflict in Afghanistan is associated with injury patterns that differ from wars such as Vietnam or Korea. This report describes the experience of an Australian military anaesthetist and the value of regional anaesthesia and analgesia for the care of the wounded combat soldier PMID- 20014612 TI - A technique that may improve the reliability of endobronchial blocker positioning during adult one-lung anaesthesia. AB - We describe a novel technique, previously applied to small children, for adult one-lung anaesthesia in which a single-lumen endotracheal tube is used with an endobronchial balloon blocker The main aims of the technique are to reduce the likelihood of cephalad displacement of the balloon into the trachea and to facilitate directional placement of the endobronchial balloon. We present five illustrative cases of one-lung anaesthesia in patients of adult size, in which the endotracheal tube-endobronchial balloon technique was considered preferable to the use of a double-lumen tube technique. The situations included difficult intubation, need for postoperative ventilation, a tortuous trachea and an unexpected need to perform one-lung anaesthesia. The technique involved deliberate placement of the endotracheal tube tip near the carina to block cephalad dislodgement of the blocker The chance of the balloon blocking the endotracheal tube tip could be further reduced by having the intraluminal endobronchial balloon blocker emerge through the Murphy eye. PMID- 20014613 TI - Brain death in a septic patient: possible relationship with posterior reversible encephalopathy syndrome? AB - A 41-year-old man with a previous kidney transplant was referred for arterial hypertension and acute renal failure. Initial neurological examination was normal. Laboratory data showed a high serum cyclosporine A concentration. A few hours later, he developed generalised tonic-clonic seizures. The brain computed tomogram was not remarkable, but Glasgow Coma Scale score remained at 8. Mechanical ventilation was required for rapidly progressive hypoxaemia related to Staphylococcus aureus pneumonia and septicaemia. Noradrenaline infusion was needed for only nine hours, with no major drop in mean arterial blood pressure. On day three his Glasgow Coma Scale score was 3/15, with fixed dilated pupils. The brain computed tomogram revealed bilateral hypodense lesions in the posterior areas together with cerebral oedema and the patient was subsequently declared brain dead. We discuss the possibility of a posterior reversible encephalopathy syndrome, likely triggered by a gram-positive septicaemia in addition to other risk factors. PMID- 20014614 TI - Intraoperative airway foreign body migration in a child. AB - We report successful anaesthetic management of a foreign body in the airway, which migrated intraoperatively from one bronchus to the other causing life threatening hypoxaemia. PMID- 20014615 TI - Entrapment of the introducing sheath of a pulmonary artery catheter. AB - We report an unusual complication of pulmonary artery catheterisation related to the removal of the pulmonary artery catheter introducing sheath. The sheath had been accidentally tethered to the suture line of the superior vena cava following cardiopulmonary bypass surgery. Physicians and nurses should abandon attempts to remove the pulmonary artery catheter or the introducing sheath after cardiac surgery if any resistance is noted. Minimal force might result in serious morbidity or death. PMID- 20014616 TI - Anaphylaxis and mastocytosis. PMID- 20014617 TI - Artifactual 'atrial flutter' caused by a continuous passive motion device after total knee replacement. PMID- 20014618 TI - Another defective Diprivan syringe. PMID- 20014619 TI - [Once again... media politics]. PMID- 20014620 TI - [Local control treatment of bacteria found present in an infected bed sore by means of a hydropolymer dressing with ionic silver]. AB - Every chronic wound has microorganisms present on its surface and therefore, its contamination is universal and unavoidable, but the majority of these wounds are not infected. It is important, therefore, to refine the diagnosis of infection indentifying the microorganisms which have succeeded in gaining access to the submarginal tissues in order to be able to prevent unnecessary or inappropriate antibiotic treatment and to lessen possible complications. A high level of bacteria present, close to 105 "UFC" per gram of tissue, even though this does not manifest a clinical infection, might interfere with the repair mechanisms and impede cicatrization of the wound. Due to this difficulty to identify in a precise manner the microorganisms which cause an infection, those patients who have sepsis related to bed sores are habitually treated empirically with wide spectrum range antibiotics, potentially more toxic and often more expensive. But it is important to restrict the use of local antibiotics when treating chronic wounds and to prioritize the use of cleaning methods and diverse debridement methods in order to lower the presence of bacteria. The use of systemic antibiotics should be limited to precise indications, considering that today we have at our disposal alternate treatments with topical antibiotics, principally dressings which contain silver. PMID- 20014621 TI - [Emotional support and Bach Flower Therapy]. AB - Bach Flower Therapy is a method to cure and prevent physical and/or emotional disorders by means of treating our emotional unbalances. Primary Health Care Centers are an excellent site to apply this therapy since their main working instrument is an interview focused on patients. Over a 14 month period, a group of patients received this type of treatment in a Primary Health Care Center in Catalonia from a nurse at the center prepared to apply this therapy. The objective of this study is to describe the patients who received it, the identified nursing diagnoses, and the evolution of the physical and/or emotional disorders after treatment has been applied. Patients were enrolled in this therapy Project based on a request by the medical/nursing consultants treating patients or by a patient's own request to participate. 119 persons, of whom 78.15% were female, received personalized therapy until their health condition improved; seven patients had been excluded from this therapy since they only received a rescue remedy. The time period between the first and the last visit varied according to each case and the individual's personality 405 visits took place, an average of 3.4 per patient. Improvement was measured on the EVA scale: 87.4% of those patients who received treatment reported good or very good evolution/results. Bach Therapy appears to be a good alternative to psycho pharmaceuticals, shows itself to be very effective, with fewer secondary effects, and less than 2% of patients evidencing gastric intolerance to this preparation. PMID- 20014622 TI - [Informed consent]. AB - At present times, numerous complaints claiming defects produced at some point in the process of obtaining informed consent are filed in courts of justice; in these complaints there is an underlying comment element which is the roles that health professionals have in these processes. In obtaining this consent, one can see this more as a means to obtain judicial protection for professional practices rather than this process being a respectful exercise for the dignity and freedom which health service patients have. This article reflects on two basic rights related to informed consent: adequately obtaining this consent and the need to protect those people who lack, either partially or totally, the capacity to make this decision by themselves. Afterwards, the author makes some considerations about the necessity to obtain informed consent for nursing practices and treatment. PMID- 20014623 TI - [Having cancer and desiring to become parents; bibliographical review]. AB - Over these recent years, survival by patients who have suffered some kinds of cancer has increased due to advances in diagnostic techniques and in treatment. Many of these patients are youths who have yet to begin to form families or have not completed them. The possibility of having a child in the future is a very important topic for cancer patients. Infertility can be associated with the disease itself or its treatments. On the other hand, the development of assisted reproduction has made it possible to achieve a large number of pregnancies. All these factors have led to recommending to cancer patients who harbor reproductive desires to use methods to preserve their fertility before starting anti-cancer treatment. In spite of these findings, studies document that oncology teams do not treat fertility with patients as frequently as they contemplate other complications. PMID- 20014624 TI - [Informative ethics: quality of services. Thoughts regarding what is going on in our communications media]. PMID- 20014625 TI - [Patients who undergo cryoplasty. Nursing results (NOC), tolerance to this operation]. AB - The medical profession has started to use a new treatment for peripheral vascular disease: cryoplasty. The objective of this article is to study the results of nursing treatment "tolerance to this operation" for patients who undergo this technique, cryoplasty. In order to know what indicators are those most adequate to quantify its evolution, the author chose these indicators: Taking a walk, Length of walk, Tolerance to climbing stairs, Ease in carrying out Daily Life Activities, Resistance by the lower body and speaking ability while exercising. The author gave a pre-and post-cryoplasty grade for each of these indicators on a 5 point Likert evaluation scale. PMID- 20014626 TI - [Patients in need of parenteral nutrition in their homes using a PICC catheter]. AB - The authors presented their information in a poster format at the VI GAVeCeLT National Congress "VI Congresso Nazionale GAVeCeLT (Centro Congressi Europa, Universita Cattolica del Sacro Cuore)". The authors point out that the use of less invasive systems, such as a PICC catheter (peripherally inserted central catheter), produce excellent results, provided the protocols for care and maintenance are correctly applied. PMID- 20014627 TI - [Female stress urinary incontinence during pregnancy. A bibliographical review]. AB - The authors have prepared a brief bibliographical review on female stress urinary incontinence during pregnancy. The high degree of incidence this disorder has and the introduction of the latest sanitary techniques combine to make necessary a profound review which aids medical professionals to realize the magnitude of this problem, as well as to diagnose and treat it effectively. Female stress urinary incontinence during pregnancy is an involuntary urine loss associated to the physical effort which an increase in abdominal pressure provokes. A literary review of articles related to this disorder leaves no doubt that among the risk factors which make a woman be predisposed to suffer female stress urinary incontinence, obstetric trauma as a consequence of pregnancy and birth is a main risk factor. The authors describe a couple of diagnostic tools, a ICIQ-SF questionnaire, and therapeutic tools, the pelvic floor muscle exercise program, whose effectiveness has been sufficiently demonstrated and may prove to have very beneficial clinical uses which can be applied in a nurse's office. PMID- 20014628 TI - [Nurses' patients' clinical record sheets; complying with legal requirements]. AB - This article describes a 100 page long descriptive study based on nurses' clinical record sheets at three hospitals to discover the degree of compliance with legal requirements. Compilation of existing laws and a specialized bibliography provided the variables for this study. These were placed inside a data base which, once the information from each clinical record was introduced, made it possible to draw conclusions. Due to the socio-linguistic context, the language used and the linguistic interferences were also analyzed. This study showed there is a high rate of non-compliance with legal requirements. The impossibility to identify the professional who fills in each record sheet, low legibility the frequent use of abbreviations, initials, and symbols, crossed out illegible words, and unfinished record sheets have been noted in this study. The results of this study pose the possibility to make available in hospitals a list of legal requirements and of abbreviations, initials, and symbols employing a unified meaning. PMID- 20014630 TI - [Andalucia is the first community to allow nurses to prescribe health products and medications]. PMID- 20014629 TI - [Can bladder catheterization in pediatrics cause complications? The case of a urethral dysuria cystograph]. AB - A urethral dysuria cystograph (CUMS (Cistografia Ureteral Miccional Seriada)) is the first diagnostic procedure, by means of x-rays, to evaluate bladder-urethral reflux. It consists of a bladder catheter to administer a radiopaque contrast dye through the ureter. To use the aforementioned technique without any asepsis measures and without knowledge about it can lead to possible complications. By means of a retrospective study using a sample of 181 patients, the authors have evaluated the incidence of possible complications and/or subsequent discomfort due to a urethral dysuria cystograph (CUMS). As final results, by means of an after test telephone call, the authors observed that 96.7% of the children who underwent this technique did not manifest any type of complication nor urinary discomfort. PMID- 20014631 TI - The clinical medicine of Hippocrates. A tool in adolescent medicine. PMID- 20014632 TI - Positive effects, side effects, and adverse events of clinical holistic medicine. A review of Gerda Boyesen's nonpharmaceutical mind-body medicine (biodynamic body psychotherapy) at two centers in the United Kingdom and Germany. AB - To review adverse events of intensive, clinical holistic medicine (CHM) as it is practiced in holistic body-psychotherapy in England and Germany. Gerda Boyesen's "biodynamic body-psychotherapy" (BBP) is an intensive type of holistic mind-body medicine used by Boyesen at two centers. METHODS: About 13,500 patients were treated during 1985-2005 period and studied for side effects and adverse events. The first author worked closely with Boyesen 1995-2005 with full insight in all aspects of the therapy and provided the data on side-effects. RESULTS: Therapy helped chronic patients with physical, psychological, sexual, psychiatric and existential problems to improve health, ability, and quality of life (NNT (number needed to treat) = 1-3). Effective in the treatment of mentally ill patients (schizophrenia, anxiety, poor mental health, low general ability). For retraumatization, brief reactive psychosis, depression, depersonalization and derealization, implanted memories, side effects from manipulations of the body, suicide/suicide attempts, hospitalization for physical and mental health problem during or 90 days after treatment, NNH (number needed to harm) > 13,500. INTERPRETATION: Intensive, holistic non-drug medicine is helpful for physical, sexual, psychological, psychiatric and existential problems and is completely safe for the patient. The therapeutic value TV = NNH/NNT > 5,000. Altogether about 18,000 patients treated with different subtypes of CHM in four different countries have now been evaluated for effects, side effects and adverse events, with similar results. PMID- 20014634 TI - Searching for health information on the Internet by undergraduate students in Phitsanulok, Thailand. AB - Searching the Internet has become an important and rapidly-expanding tool for users with health concerns. OBJECTIVES: This study was designed to describe the accessibility, searching pattern, and implication for obtained health information on the Internet of Thai undergraduate students. METHOD: Questionnaires composed of personal data and accessibility to drug information, were provided to convenience samples of undergraduate students in Phitsanulok. RESULTS: Four hundred questionnaires were completed, 66% females, mean age (SD) 21.58 (2.26) years old (range 17-29). Ninety percent of respondents accessed the Internet; 75% percent had access to the Internet at school in range of 1-3 times per week and 1 3 hours per time. Half the Internet users had access to health information. The most sought health information included general health (65.7%), disease and treatment (49.5%), and nutrition (41.9%). The main reasons for searching on the Internet were easy access (38.5%) and updated information (34.6%). Seventy percent of the Internet-based information seekers used the obtained information in their daily life. CONCLUSION: This study summarizes current Internet searching patterns of undergraduate students in Thailand. The results have implications for constructing realistic simulations of searching behavior and improving the value of websites on health information relevant to youth. PMID- 20014633 TI - Premarital sex and condom use among never married youth in Vietnam. AB - Premarital sex and unprotected sexual intercourse are issues of growing concern in Vietnam. The present study aims to explore the factors associated with the onset and delay of premarital sex among youth in Vietnam. METHODS: A cross sectional in-home survey was undertaken in a suburb of Hanoi, Vietnam among a sample of 2,394 never married youth ages 15 to 24 years in the end of 2003. RESULTS: Cox regression analyses showed that youth who had ever experienced sexual abuse were 35 times more likely to report having had premarital sex. Connectedness to mother likewise was associated with a decreased likelihood of having sex before marriage. A medium level of school connectedness was also a factor associated with the delay of premarital sex (OR = 0.27). Peer social deviance, on the other hand, increased the likelihood by at least 2.6 times. Separate models for each sex confirmed: the protective effect of mother connectedness among females; the negative effect of peer social deviance among males; and the predominant risk of past sexual abuse in both sexes for premarital sex. When factors associated with condom use among sexually active never married youth were explored, males were found more likely to use condom at first sexual intercourse. CONCLUSIONS: These results reaffirm a growing body of literature from around the world that has shown connectedness to a parent to be associated with delay of premarital sex, while peer social deviance and exposure of previous sexual abuse have been both associated with early sexual debut. PMID- 20014635 TI - Sexuality and persons with Down syndrome. A study from Brazil. AB - In recent years, important gains and changes have been observed in the life of teenagers with Down syndrome (DS) with increased inclusion into society. This review will discuss adolescence and sexuality in teenagers with DS from a descriptive study of 50 patients with DS between the ages of 10 and 20 years. The mean age was 13.5 years, 50% females; 86% went to school with 62.2% in school for over six years. Of the patients that attended school, 60% went to special education school and only 10% read and wrote correctly. In an evaluation of autonomy, 66% took showers, 78% performed their physiological needs, 77% intimate hygiene and 76% oral hygiene without help. 42% affirmed being able to do anything that is asked; 22% perform all tasks in the home; 10% felt they were incapable of doing anything and 4% used public transportation without help. 42% of the teenagers masturbated, 24% on a daily basis, 75% in private, and 25% in a public location. 42% had already kissed at a mean age of 12.9 years, mean age of the partner 16.1 years; 26.8% of these partners had DS. 82% found themselves attractive and 33% would not change anything in their appearance. We found that they presented normal development in the exercise of their sexuality, but with important difficulties in their autonomy and difficulties in school, needing careful interventions to make their social interaction the best possible. Their pubertal development was normal and they were satisfied with their body image with future perspectives of working, finding a partner, and living a normal life of getting married and having children. PMID- 20014636 TI - Self-related health, physical activity, BMI and musculoskeletal complaints: a comparison between foreign and Swedish high school students. AB - Physical activities during leisure time and school hours as well as nutritional habits have changed over the past years by adolescents being less physically active and adopting a sedentary life-style. The aim of the present study was to investigate possible differences between foreign and Swedish high school students in terms of self-related health, physical activity, overweight, and possible complaints from the musculoskeletal system. METHODS: 1,090 high school students, 450 with foreign background and 640 with Swedish background, aged 16-26 years answered a questionnaire. RESULTS: A higher percentage of students with foreign background reported poor self-related health compared with students with Swedish background (p = .038). Students with a foreign background were to a greater extent less physically active than students of Swedish background (p = .003). No differences were found between the groups regarding musculoskeletal complaints. Students with foreign background were more often overweight than students with Swedish background and overweight was more frequent among males than females. Physical activity (moderate and high level) was concluded to be a factor with significant positive effect on self-related general health (moderate level p = .042, high level (p < .001), and musculoskeletal complaints were negative factors on self-related general health (p < .001). CONCLUSION: The results suggest that adolescents with foreign background should participate in physical activity to prevent overweight and thereby improve physical health. PMID- 20014637 TI - Screening Belgian university students for Chlamydia trachomatis infection: a feasibility study. AB - In this study we examine the attainability and usefulness of opportunistic screening for Chlamydia trachomatis infection based on self-assessed risk, among university students in Belgium. METHODS: A self-administered questionnaire was filled out by students (n = 243, 77.8% female, 22.2% male), who were asked to assess their own risk, to decide if their participation was useful, and to collect a first-void urine sample. Specimens were refrigerated and delivered to the laboratory on the same day. Screening for C. trachomatis DNA was performed by PCR. A Positive result was confirmed by another amplification assay. RESULTS: Two hundred forty three students took part in the study. One hundred thirteen participants did not meet the inclusion criteria. Ages varied from 18 to 39 years, with a mean age of 21.49 years. The overall prevalence of C. trachomatis infection was 2.9%. The prevalence of C. trachomatis infection in the group of students who met the inclusion criteria was 5.4%. Having a new partner in the past six months and having had more than one partner in the last year were the most frequent reported risk factors in male and female participants. CONCLUSIONS: Screening students is a useful and feasible strategy to diagnose asymptomatic chlamydial infection. However assessing their own risk of infection seemed difficult for students. The overscreening of youngsters not at risk and the limited participation of males should get extra attention. PMID- 20014638 TI - Exploring the neighborhood: a web-based survey on the prevalence and determinants of fear among young adolescent girls and boys. AB - Children's independent everyday mobility can be hindered by fears experienced in their neighborhood. The aim of this study is to assess the prevalence and determinants of such fears among boys and girls in early adolescence, a period when individual freedom is expected to be on the increase. A sample of 7th grade students (age-13 years) in Stockholm County, Sweden, during 2005/06 answered a survey in class (n = 1,008). The relation that gender, housing, family characteristics, individual and peer negative experiences in the neighborhood, parental licensing, and length of stay in the neighborhood have with fear disclosure was assessed through multivariate logistic regression. A total of 60% of the girls and 40% of the boys reported experiencing fears in their neighborhood. Gender differences were significant for all of the most common fears, in particular darkness. When respondents or their friends had been chased, hit, or had something taken from them in their neighborhood, they were more likely to report fear (OR girls 2.3; 95% CI 1.6-4.5; boys 2.8; 95% CI 1.9-4.2). For girls, having one or more parents born outside Sweden was associated with fear. Boys nearly three times more often reported fear if (a) they thought their parents were negative toward adolescent independent mobility in the evening, or (b) they had lived longer than one year in their area. Many young adolescents admitted to experiencing fear in their neighborhood. Fears were more common among girls, and the types and determinants of fear seem to be gender specific. PMID- 20014639 TI - Medication taking behavior of students attending a private university in Bangladesh. AB - The objective of this study was to investigate the patterns of self-reported medication use, including both prescription and OTC drugs, and to assess the possible predictors of self-medication and medication non-compliance (non adherence), for 929 non-medical undergraduate students of the American International University, a private university situated in Dhaka city, Bangladesh. Although a high proportion (69%, n = 644) of students of this university had fallen sick in the last six months before the study, the rate of visiting qualified health practitioners was much lower (53%). A good proportion of the sick students were reported to have practiced self-medication (16%, n = 100) and medication non-adherence (15%, n = 98). The average treatment cost involved in self medication was much lower than that offered by a qualified physician (Tk 463 vs Tk 2546 per case). Those students living with parents were more likely to have visited qualified health practitioners (56%, p < .05), and students whose families kept a well-stocked medicine cabinet at home were more likely to have completed the full course (39%, p < .05) of prescribed medicine. No significant difference was found in the rates of self medication and medication compliance incidence for variables like age groups, gender, residence status, financial level, engagement in part-time jobs etc. The study also showed that antimicrobials are widely available (170 incidents) in the home medicine cabinets of the Dhaka City population. The storage of leftover antibiotics in the home constitutes an alternative potential source of self-medication that can have untoward consequences. Further elaborate studies are required to reveal the true pattern of antibiotic usage in Bangladesh. PMID- 20014640 TI - A longitudinal analysis of teenage drinking patterns. AB - This paper examines the multinomial recurrent outcome 'teenage drinking' within a statistical modeling paradigm. The proposed statistical modeling relates drinking to a set of explanatory variables, which include subjective as well as objective measures. In order to assess the degree to which explanatory variables influence smoking, an adequate statistical model must handle the possibility that substantial variation between respondents will be due to omitted variables, multicollinearity and past behavior. To address these issues, longitudinal data using appropriate statistical modeling are essential. In this paper we demonstrate the application of appropriate longitudinal modeling using data from the Yorkshire (UK) Health Related Behavior Questionnaire. The results suggest there may be substantial heterogeneity due to omitted variables in the data and complex inter-relationships between observed explanatory variables. For example, social cultures of drinking and parental involvement appear to influence heavy drinking and drug use. PMID- 20014641 TI - Sexual behavior and risk of HIV/AIDS among adolescents in public secondary schools in Osogbo, Osun State, Nigeria. AB - Young people are particularly vulnerable to unplanned sexual activities. This study sought to identify the sexual behaviors and risk of HIV among public secondary schools students in Nigeria. METHODS: A cross-sectional survey of 521 students in eight randomly selected public secondary schools. Respondents were selected by a multistage sampling technique from amongst the study population. Using a self-administered, semi-structured, pre-tested questionnaire, data were obtained concerning their sexual behaviors and influencing factors. RESULTS: Most respondents (n = 387, 74.3%) were in late adolescence. Many knew the consequences of premarital sex as unplanned pregnancy, STI/HIV/AIDS, incomplete schooling, and guilt feelings. About 40% of the respondents had been involved in sexual activities with partners who were classmates, neighbors, 'sugar daddies', teachers, or strangers (party-mates or prostitutes). Heterosexual, oral, and anal forms of sex were practiced respectively by 78.1%, 13.3%, and 12.4% of those who were sexually active. Sexual debut was 15.2 +/- 1.62 years. About 36% of those sexually active had more than one partner, and about 14.8% were aware that their partners had other partners. Only 8.6% used a condom on a consistent basis, whereas 41.9% had never used a condom at all. More than half the sexual activities were not pre-planned. The reasons given for engaging in such practices were peer influence, financial reward, drug influence, fun, or experimentation. CONCLUSIONS: Despite their 'above average' level of knowledge of the consequences, the students were still involved in risky sexual behaviors. Behavioral change communications should be intensified among these adolescents. PMID- 20014642 TI - Evaluation of school-based reproductive health education program for adolescent girls. AB - AIM: To evaluate the effectiveness of school-based reproductive health education for adolescent girls on the reproductive knowledge level of the girls. METHODS: This research was carried out as a quasi-experimental study at two vocational girls high schools, one of which was used as the study school and the other as the control school. The study group (97 students) consisted of three classes representing every grade. The control group consisted of students selected likewise (92 students). Reproductive health education was given to students in the study group for 10 weeks; the control group was not subjected to any educational program. The impact of the program was evaluated with reproductive health knowledge test designed for this study. A pretest evaluated baseline knowledge, and a posttest measured the gain in knowledge. RESULTS: Baseline knowledge score of students in study and control group were similar and low (p > 0.05). We found that the reproductive health knowledge level of students in the study group increased significantly after the program of education. Post-test knowledge scores (75.03 +/- 13.82) of the students in the study group were higher than those of the control group (36.65 +/- 14.17). CONCLUSIONS: The results showed students' low baseline knowledge and a good ability to learn. A school based reproductive health education is needed to promote knowledge and prevention in reproductive health among teenagers. PMID- 20014643 TI - Contraceptive knowledge and practice: a survey of under graduates in Ibadan, Nigeria. AB - Despite the mounting reproductive health problems among adolescents in Nigeria, very little has been done at governmental level to address the problem, hence presenting a major health concern and a leading cause of morbidity and mortality. OBJECTIVE: To determine the knowledge, attitude of the undergraduates to contraception and their practice of the various contraceptive options. METHODS: A descriptive cross-sectional study was done among the undergraduates of the Polytechnic of Ibadan and University of Ibadan, Nigeria. Interviewer-administered questionnaires were given to 600 consenting undergraduates. The data were analyzed using SPSS version 11 software. Chi2 and Fisher exact tests were used for categorical variables as appropriate. RESULTS: A large percent (65.1%) of the undergraduates were sexually experienced. Approximately, 90.2% of the respondents who had sex did so before the age of 19 years, 31.8% had more than one sexual partner, and 21.8% had clandestine abortions. All respondents were quite knowledgeable about HIV/AIDS but few (30.2%) of the sexually active took precautions to prevent HIV transmission. The majority (65.4%) were not knowledgeable about contraception, and 10.5% heard about contraception from parents. About 52.3% of the latter cited pharmacy shops as their source of contraceptives. CONCLUSION: The attitudes of the students were below expectations. Specially designated centers for the provision of appropriate contraceptive services to students by trained personnel are needed in the institutions to tackle their reproductive health problems. PMID- 20014644 TI - Personality characteristics and profiles of Greek elementary teachers using the sixteen personality factor questionnaire (16PF). AB - Empirical evidence indicates that a teacher's personality influences the classroom climate, students' behaviors, and their interpersonal relationships. Although the effect of a teacher's personality on students' psychological well being has long been stressed in many studies, very little is known about the actual personality characteristics of Greek in-service teachers. The purpose of this study was to allocate the characteristics that best describe the personality of Greek elementary school teachers (according to the 16 Cattellian primary factors). Our study belongs in the broader research field aiming at describing and understanding the possible foundations of teachers' behavior. The sample consisted of 138 elementary teachers, who completed a standardized Greek version of the 16PF. Our statistical analysis of one-sample t-test along with an effect size calculation revealed that certain personality characteristics described the Greek elementary teacher and clearly distinguishes them from the normative group of the Greek population. Elementary teachers appear to be quite submissive, cautious, with a tendency to oppose or postpone change. They also scored a low tolerance level against fear and arousal, and high tension levels. Elementary teachers seem to respond to events, ideas, and experiences more with feeling than with thinking and find it difficult to control their feelings, which results in getting upset easily. They also seem to pay little attention to how they may appear to others and generally do what they feel like doing. Elementary teachers also scored low on aspiration level. Possible implications of the results are discussed with reference to students' psychological well-being. PMID- 20014645 TI - Residential care centers for persons with intellectual disability in Israel. Trends in the number of nursing care patients during 1998-2007. AB - Nursing care has changed over time and the focus has differed according to the need or the culture involved. One of the first descriptions of nursing of people with intellectual disability (ID) is from the United States at the New York Hospital around 1808. Over the next 200 years, ID nursing developed into a subspecialty in both the United States and England with a curriculum and certification. This process has yet to take place in Israel, but it is a change we would like to see in the coming years. The purpose of this paper was to look at the trend in the number of nursing patients in residential care centers for people with ID in Israel. Data were extracted from the 1998-2007 national annual surveys of all residential care centers in Israel. Over this 10-year period, we found that the number of nursing patients was fairly constant, ranging from 23.9% of the total population in 1998 to 26.5% in 2007. Whereas the percent of nursing patients has remained steady, the burden of nursing has increased. Specifically, due to the aging of this population, residential care centers are now dealing with an increased quantity and complexity of medical problems. As a result, we have experienced a need to establish, on a regional basis, long-term nursing care facilities within our residential care centers. PMID- 20014646 TI - Context-dependent modulation of cutaneous reflex amplitudes during forward and backward leg cycling. AB - We used amplitude modulation of cutaneous reflexes during leg cycling as a paradigm to investigate neural control mechanisms regulating forward (FWD) and backward (BWD) rhythmic limb movement. Our prediction was a simple reversal of reflex modulation during BWD leg cycling and context-dependent reflex modulation. Cutaneous reflexes were evoked by electrical stimulation delivered to the superficial peroneal (SP) and distal tibial (TIB) nerves at the ankle. EMG recordings were collected from muscles acting at the hip, knee, and ankle. Kinematic data were also collected at these joints. Cutaneous reflexes were analyzed according to the phase of movement in which they were evoked. When functional phases (i.e., flexion or extension) of cycling were matched between FWD and BWD, background EMG and reflex modulation patterns were generally similar. The reflex patterns when compared at similar functional phases presented as a simple reversal suggesting FWD and BWD cycling are regulated by similar neural mechanisms. The general reflex regulation of limb trajectory was maintained between cycling directions in accordance with the task requirements of the movement direction. PMID- 20014647 TI - Muscle activation patterns and postural control following stroke. AB - Many stroke survivors have residual sensorimotor deficits that impact negatively on balance and quality of life. The purpose of this review is to provide an overview of the impairments in motor control following stroke and the impact of those impairments on muscle activation patterns during postural control in stroke. Motor control impairments following stroke result in force production that is slow, weak and lacking in precision making it difficult to produce a fast rate of force development with sufficient magnitude to be effective for postural responses. Whether postural perturbations require feedback or feedforward responses, there is impairment to the timing, magnitude and sequencing of muscle activation following stroke. The impairment in muscle activation is dependent on the extent of the motor control impairments and strategies used by the individuals following stroke to compensate for the impairments. The central nervous system uses a variety of mechanisms to improve the muscle activation patterns needed for the recovery of postural responses following stroke. PMID- 20014648 TI - Brief electrical stimulation accelerates axon regeneration in the peripheral nervous system and promotes sensory axon regeneration in the central nervous system. AB - Injured peripheral but not central nerves regenerate their axons but functional recovery is often poor. We demonstrate that prolonged periods of axon separation from targets and Schwann cell denervation eliminate regenerative capacity in the peripheral nervous system (PNS). A substantial delay of 4 weeks for all regenerating axons to cross a site of repair of sectioned nerve contributes to the long period of separation. Findings that 1h 20Hz bipolar electrical stimulation accelerates axon outgrowth across the repair site and the downstream reinnervation of denervated muscles in rats and human patients, provides a new and exciting method to improve functional recovery after nerve injuries. Drugs that elevate neuronal cAMP and activate PKA promote axon outgrowth in vivo and in vitro, mimicking the electrical stimulation effect. Rapid expression of neurotrophic factors and their receptors and then of growth associated proteins thereafter via cAMP, is the likely mechanism by which electrical stimulation accelerates axon outgrowth from the site of injury in both peripheral and central nervous systems. PMID- 20014649 TI - Measurement and modulation of plasticity of the motor system in humans using transcranial magnetic stimulation. AB - Several techniques that involve transcranial magnetic stimulation (TMS) can be used to measure brain plasticity noninvasively in humans. These include paired associative stimulation (PAS), repetitive transcranial magnetic stimulation (rTMS) and theta burst stimulation (TBS). Some of these techniques are based the principle of use dependent plasticity or are designed to mimic protocols used to induce long-term potentiation or depression in animal studies. These studies have been applied to certain neurological and psychiatric disorders to investigate their pathophysiology. For example, PAS induced plasticity is enhanced in dystonia and stroke but is reduced in Huntington's disease and schizophrenia. Furthermore, TMS may be used to modulate brain plasticity and has therapeutic potential in neurological and psychiatric disorders such as stroke, Parkinson's disease, dystonia and depression. PMID- 20014650 TI - Modulation of arm stiffness in relation to instability at the beginning or the end of goal-directed movements. AB - In reaching to a target, stability near the target may be more critical for success than stability far from the target. Consequently, we postulated that high instability near the start would evoke less compensation than high instability near the target. Three stability conditions were implemented using a robot manipulandum: neutral stability everywhere (null field); high instability along the first half of the trajectory decreasing as the target was approached (start unstable); and instability increasing along the first half of the trajectory and remaining high as the target was approached (end unstable). Under the start unstable condition, the stiffness of the arm in the region of highest instability was significantly less than under the end unstable condition. Furthermore, the stability of the system (manipulandum plus arm) was much lower under the start unstable condition than under the null field condition whereas it was similar under the end unstable and null field conditions. PMID- 20014651 TI - Interpersonal postural coordination on rigid and non-rigid surfaces. AB - When two standing people converse with each other there is an increase in their shared postural activity, relative to conversation with different partners. We asked pairs of participants to converse with each other or with experimental confederates while standing on rigid and nonrigid surfaces. On the rigid surface, shared postural activity was greater when participants conversed with each other than when they conversed with confederates. In addition, the strength of interpersonal coupling increased across trials, but only when members of a dyad conversed with each other. On the nonrigid surface, postural sway variability increased, but we found no evidence that shared postural activity was different when participants conversed with each other, as opposed to conversing with confederates. We consider several possible interpretations of these results. PMID- 20014652 TI - Proteomic profiling of the rat cerebral cortex in sleep and waking. AB - Transcriptomic studies have shown that hundreds of genes change their expression levels across the sleep/waking cycle, and found that waking-related and sleep related mRNAs belong to different functional categories. Proteins, however, rather than DNA or RNA, carry out most of the cellular functions, and direct measurements of protein levels and activity are required to assess the effects of behavioral states on the overall functional state of the cell. Here we used surface-enhanced laser desorption-ionization (SELDI), followed by time-of-flight mass spectrometry, to obtain a large-scale profiling of the proteins in the rat cerebral cortex whose expression is affected by sleep, spontaneous waking, short (6 hours) and long (7 days) sleep deprivation. Each of the 94 cortical samples was profiled in duplicate on 4 different ProteinChip Array surfaces using 2 different matrix molecules. Overall, 1055 protein peaks were consistently detected in cortical samples and 15 candidate biomarkers were selected for identification based on significant changes in multiple conditions (conjunction analysis): 8 "sleep" peaks, 4 "waking" peaks, and 4 "long sleep deprivation" peaks. Four candidate biomarkers were purified and positively identified. The 3353 Da candidate sleep marker was identified as the 30 amino acid C-terminal fragment of rat histone H4. This region encompasses the osteogenic growth peptide, but a possible link between sleep and this peptide remains highly speculative. Two peaks associated with short and long sleep deprivation were identified as hemoglobin alpha1/2 and beta, respectively, while another peak associated with long sleep deprivation was identified as cytochrome C. The upregulation of hemoglobins and cytochrome C may be part of a cellular stress response triggered by even short periods of sleep loss. PMID- 20014653 TI - Diffusion tensor MRI and MR spectroscopy in long lasting upper motor neuron involvement in amyotrophic lateral sclerosis. AB - Upper motor neuron (UMN) dysfunction in Amyotrophic Lateral Sclerosis (ALS) is not easy to identify clinically: Diffusion Tensor Imaging (DTI) and single-voxel Magnetic Resonance Spectroscopy (H-MRS) can identify markers of UMN involvement. The aim of this study was to correlate brain DTI and MRS data with clinical parameters in ALS patients (PALS). We studied 32 PALS using Magnetic Resonance Imaging. The subjects were subdivided into definite/probable (D/P) and possible/suspected (P/S). DTI indices included Fractional Anisotropy (FA) and averaged Apparent Diffusion Coefficient (avADC). Anatomical areas were sampled by positioning regions of interest along corticospinal tracts, from the precentral cortex to the bulb. H-MRS voxels were localized bilaterally in precentral regions. D/P-PALS showed significantly lower FA values than healthy controls in almost all regions, whereas P/S-PALS FA values were significantly lower only in the left precentral gray matter (GM), right precentral white matter (WM), cerebral peduncles (CP), left hemipons, and left bulbar pyramid (BP). Significantly higher avADC values were observed in the D/P-PALS right precentral GM, precentral WM, right semioval center-posterior limb of the internal capsule (SC-PLIC), and left CP; and in the precentral WM, right SC-PLIC, left CP, and right hemipons of P/S-PALS. With increasing disability, only D/P-PALS showed significantly reduced FA values in the left precentral WM and hemipons, and increased avADC values in the precentral WM. Significantly lower N acetylaspartate (NAA)/creatine-phosphocreatine complex (Cr) and higher choline (Cho)/Cr and myoinositol (mI)/Cr ratios were found in D/P-PALS, while only higher Cho/Cr and mI/Cr ratios were found in P/S-PALS. Our data highlight the usefulness of DTI and H-MRS in assessing UMN involvement. Given FA sensitivity and specificity, despite the small number of PALS, our findings support its use as a diagnostic marker in D/P-PALS. PMID- 20014654 TI - Changes in medial prefrontal cortex neural responses parallel successful antidepressant combination of venlafaxine and light therapy. AB - Few pilot prospective studies performed BOLD fMRI before and after treatment in order to define the neural correlates of antidepressant response. To determine how antidepressant treatment influences the pattern of neural response to a task targeting the depressive biases in information processing (moral valence decision), eight depressed inpatients were treated with combined venlafaxine and light therapy for four weeks. Brain BOLD functional magnetic resonance imaging on a 3.0 Tesla scanner was performed before and after treatment. Treatment and moral value of the stimuli showed the most significant interaction in right medial frontal gyrus (BA 10), where also clinical status was found to be inversely correlated with response to negative stimuli after treatment. A significant interaction of treatment and valence of the stimuli was also detected in other areas that have been widely associated with the depressive illness. PMID- 20014655 TI - Be(a)ware of spider! An attentional blink study on fear detection. AB - We investigated whether detection of fearful stimuli is independent from attention by using an iconic version of the Attentional Blink Task in arachnophobic individuals. A colored animal icon (Target) and a black spider or butterfly icon (Probe) appeared in close temporal proximity within a stream of distractors, at one of 4 possible time lags. In one task, Probe detection was required; in another one, Target identification was also requested. In this case, competition for attentional resources produces the so-called AB effect, that is the decrease of Probe perception as a function of lag. During spider-Probe detection, arachnophobics showed a reduced AB effect with respect to the butterfly-Probe session. Their spider detection scores were also greater than ratings obtained by non-phobic controls with both Probe types. Thus, fear appears to enhance the probability of consciously perceiving the stimulus even when attention is engaged by a previous demanding event. One may assume that spider Probe is scarcely attention demanding because detection of threat in arachnophobics is increased by rapid amygdala activation of visual areas and/or facilitated by a strong arousal-induced noradrenergic cortical input. Alternatively, an attention capturing mechanism involuntary triggered by the phobic meaning of the stimulus could be hypothesized. PMID- 20014656 TI - Use our name! PMID- 20014657 TI - Patient information. Off to college? Health issues to remember. PMID- 20014658 TI - Game on! Guidance for performing PPEs. PMID- 20014659 TI - Sports physicals in the retail setting. PMID- 20014661 TI - Forced spirometry: an underused but useful tool. PMID- 20014660 TI - Three generations and counting. PMID- 20014662 TI - Determining glycemic control. PMID- 20014663 TI - The power of the PDA. PMID- 20014665 TI - Looking back, staying strong. PMID- 20014666 TI - Understanding liability insurance. PMID- 20014667 TI - Interphalangeal joint reduction. PMID- 20014668 TI - Patient information. Treatment options for hearing loss. PMID- 20014669 TI - Accidental entrepreneur. KC Arnold, NP: Entrepreneur of the Year. Interview by Jill Rollet. PMID- 20014670 TI - One to watch: Juliet Santos, NP. An NP's ambitious vision rises above the crowd. Interview by Sarah Lebo. PMID- 20014671 TI - Menace in the mix. Avoiding drug interactions with food, herbs and nutrients. PMID- 20014673 TI - Taking charge. NPs strategize in the face of opposition. PMID- 20014672 TI - Renewing relationships. Treating sexual pain in women. PMID- 20014674 TI - What now? Responding to relapse in hepatitis C. PMID- 20014675 TI - Suicide in older adults. A priority concern. PMID- 20014676 TI - The primary target. Taking aim at LDL levels to prevent cardiovascular disease. PMID- 20014677 TI - Writing a physical therapy prescription. Key components for optimum outcomes. PMID- 20014679 TI - Benefits of reciprocity. It's hard to ask for help. PMID- 20014678 TI - Outer limits. An overview of otomycosis. PMID- 20014680 TI - Patience for patients. PMID- 20014681 TI - Enjoying the holidays, blood sugar in check. PMID- 20014683 TI - Promoting testicular health. PMID- 20014684 TI - The management of type 2 diabetes mellitus: a novel approach for addressing glycemic and lipid control with colesevelam HCl. PMID- 20014685 TI - The 'liquid' facelift. PMID- 20014686 TI - Treating type 2 diabetes. A specialist's approach. PMID- 20014687 TI - Diabetes in the retail clinic setting. PMID- 20014688 TI - HPV prevention update. New vaccine offers additional options. PMID- 20014689 TI - HPV vaccination for adolescents. An ethics case study. PMID- 20014690 TI - Stress-induced cardiomyopathy in women. PMID- 20014691 TI - Mild cognitive impairment. Routine screening can slow the course. PMID- 20014692 TI - Incontinence in older adults. PMID- 20014693 TI - Anaphylaxis alert. PMID- 20014694 TI - 'I wish I could...'. Strategies for learning new skills. PMID- 20014695 TI - From grocery cart to cellulitis. A reminder about everyday health threats. PMID- 20014696 TI - Sweet dreams. PMID- 20014697 TI - SNF billing clarified. PMID- 20014698 TI - Privacy issues. HIPAA and CMS provide guidance. PMID- 20014699 TI - Colorectal cancer. Risk, screening and referral. PMID- 20014700 TI - Small price for a big dream. NP Entrepreneur of the Year: Teri Bunker, NP. PMID- 20014701 TI - One to watch: Tanya R. Sorrell, NP. Sweetening the sting. PMID- 20014703 TI - Evaluation of cough in adults. PMID- 20014702 TI - Outside the box: Renee Mercer, NP. Tackling an 'alarming' problem. PMID- 20014704 TI - Botulinum toxin A. An alternative treatment for migraine. PMID- 20014705 TI - Addressing a bad review. Offer a mature and professional response. PMID- 20014706 TI - Beyond the clinical. Ethics study can improve practice. PMID- 20014707 TI - Hanging tough in today's economy. PMID- 20014708 TI - The NICU: first site of acute care. PMID- 20014709 TI - More visibility. Retail health improves NPs' forecast. PMID- 20014710 TI - Patient information. Protein's role in your diet: food sources are best. PMID- 20014711 TI - Hypertension management. The cornerstone of heart disease prevention. PMID- 20014712 TI - Hypertension management in a retail setting. PMID- 20014713 TI - Improving cardiac outcomes and patient satisfaction. NP practice in a community hospital. PMID- 20014714 TI - Depression in Chinese immigrants. Overcoming the cultural stronghold on symptom report. PMID- 20014715 TI - Communication is the goal. Helping parents of deaf children. PMID- 20014716 TI - Palliative sedation at the end of life. Exploring a conflict of duty. PMID- 20014717 TI - Medicare's e-prescribing incentive program. An introduction. PMID- 20014718 TI - Referring to another NP. It's more than who you know. PMID- 20014719 TI - When the time comes. Retirement can sneak up on you. PMID- 20014721 TI - Sex differences in gallbladder cancer. PMID- 20014722 TI - Nutritional supplements. Good for your skin, too. PMID- 20014723 TI - Beneath the surface. Psoriasis is more than skin deep. PMID- 20014724 TI - Low back pain in a retail clinic. PMID- 20014726 TI - Metabolic syndrome in Native Americans. PMID- 20014725 TI - Saving face. Strategies to fight father time. PMID- 20014727 TI - Your first NP position. PMID- 20014728 TI - Wrong foot forward. Cautionary tales in job hunting. PMID- 20014729 TI - Finding answers. Make sure your practice is legal. PMID- 20014730 TI - Professional liability insurance. How much do you need? PMID- 20014731 TI - Sticks and stones. It matters what others call you. PMID- 20014732 TI - Mastering a techie trick. PMID- 20014733 TI - Signs of life. PMID- 20014734 TI - First steps to success. Meeting the requirements for practice. PMID- 20014735 TI - Sex differences in obesity. Genetic and hormonal causes at work. PMID- 20014736 TI - Creating a practice Web site. From ISP to URL, here's what you need to know. PMID- 20014737 TI - Pain and anemia after a gunshot wound. Pain syndrome or lead toxicity? PMID- 20014738 TI - Treatina genital warts. Application of trichloroacetic acid. PMID- 20014739 TI - Patient information: Pumping breast milk for baby. A patient guide to expressing milk. PMID- 20014740 TI - A decade of growth. Salaries increase as profession matures. PMID- 20014741 TI - The uninvited guest. An overview of multiple sclerosis. PMID- 20014742 TI - Gastroesophageal reflux in infancy. A common but challenging presentation. PMID- 20014743 TI - Two weddings and a funeral. Everlasting love. PMID- 20014744 TI - The breast cancer navigator. A new role for nurse practitioners. PMID- 20014745 TI - Erectile dysfunction. A predictor of metabolic syndrome? PMID- 20014746 TI - Rituximab for rheumatoid arthritis. Practical guidance for optimizing treatment. PMID- 20014747 TI - Prehypertension. Why should we worry? PMID- 20014748 TI - Toot your own horn. Don't be afraid to take credit earned. PMID- 20014749 TI - 'Nurse' or 'Nurse practitioner'? Our roots run deep. PMID- 20014750 TI - Modulation of the mitochondrial cytochrome bc1 complex activity by chromanols and related compounds. AB - Tocopherols (alpha-, beta-, gamma-, and delta-Toc) and tocopheryl quinones (alpha , beta-, gamma-, and delta-TQ) were recently suggested to modulate mitochondrial electron transfer in mammals. Intriguingly, Tocs and stigmatellin, a potent inhibitor of the mitochondrial cytochrome (cyt) bc(1) complex, possess a common structural feature: the chroman core. Therefore, we studied the interference of Tocs as well as synthetic model compounds (low molecular weight TQ analogues and tetramethyl chromanones) at the mitochondrial cyt bc(1) complex. Enzymatic experiments revealed that besides the inhibitor stigmatellin, among natural vitamin E-related derivatives, gamma-TQ/delta-TQ and, among synthetic compounds, TMC2O (6-hydroxy-4,4,7,8-tetramethyl-chroman-2-one) were most effective in decreasing the cyt bc(1) activities. Stopped-flow photometric and low-temperature electron paramagnetic resonance spectroscopic experiments showed for TMC2O an inhibition of electron transfer to cyt c(1) and a modulation of the environment of the Rieske iron-sulfur protein (ISP). Docking experiments suggest a binding interaction of the 6-OH group and 1-O atom/2-C( horizontal lineO) group of TMC2O with Glu-271 (cyt b) and His-161 (ISP) in the cyt bc(1) complex, respectively. This binding pose is similar but not identical to the potent inhibitor stigmatellin. The data suggest that chroman-2-ones are possible templates for modulatory molecules for the cyt bc(1) target. PMID- 20014751 TI - Influence of substrate complexity on the diastereoselective formation of spiroiminodihydantoin and guanidinohydantoin from chromate oxidation. AB - Chromate is a human carcinogen with a poorly defined mechanism of DNA damage. In vitro and prokaryotic studies have shown that DNA damage may occur via the formation of the hydantoin lesions guanidinohydantoin (Gh) and spiroiminodihydantoin (Sp) from further oxidation of 8-oxo-7,8-dihydroguanine (8oxoG). The unusual structure of these lesions coupled with their enhanced mutagenicity make them attractive for study with regard to their role in chromate induced cancer. We have studied the formation of Gh versus Sp and their associated diastereomers following oxidation by model Cr(V) complexes and from in situ chromate reduction by ascorbate and glutathione. Identification of the two optically assigned diastereomers of Sp (R-Sp and S-Sp) as well as the two diastereomers of Gh (Gh1 and Gh2, not yet optically assigned) was carried out using increasingly sterically hindered substrates (nucleoside --> ssDNA --> dsDNA). Lesion formation and diastereomeric preference were found to be highly oxidant- and substrate-dependent. The Ir(IV)-positive control showed a shift from near equal levels of Gh and Sp and near equal levels of all four diastereomers in the nucleoside to all Gh formation in dsDNA, with a 5-fold enhancement in Gh2 over Gh1. The two model Cr(V) complexes used in this study, Cr(V)-salen and Cr(V) ehba, showed opposite trends going from nucleoside to dsDNA with Cr(V)-salen giving enhanced Sp formation (with mainly R-Sp formed) and the Cr(V)-ehba having an oxidation profile nearly identical to that of Ir(IV). The two chromate reduction systems, Cr(6+)/ascorbate and Cr(6+)/glutathione, designed to model the intracellular reduction of chromate, showed lower levels of oxidation in all substrates. Notable in this group was the shift in the formation of the lesions to essentially all Sp for the Cr(6+)/ascorbate system with the most sterically hindered substrate, dsDNA. These results, when coupled with the known diastereomeric preference for excision of hydantoin lesions by the hNEIL1 enzyme, show the importance of defining both levels of lesion formation and diastereomeric preference of formation with regard to their potential impact on chromate carcinogenesis. PMID- 20014752 TI - Cheminformatics analysis of assertions mined from literature that describe drug induced liver injury in different species. AB - Drug-induced liver injury is one of the main causes of drug attrition. The ability to predict the liver effects of drug candidates from their chemical structures is critical to help guide experimental drug discovery projects toward safer medicines. In this study, we have compiled a data set of 951 compounds reported to produce a wide range of effects in the liver in different species, comprising humans, rodents, and nonrodents. The liver effects for this data set were obtained as assertional metadata, generated from MEDLINE abstracts using a unique combination of lexical and linguistic methods and ontological rules. We have analyzed this data set using conventional cheminformatics approaches and addressed several questions pertaining to cross-species concordance of liver effects, chemical determinants of liver effects in humans, and the prediction of whether a given compound is likely to cause a liver effect in humans. We found that the concordance of liver effects was relatively low (ca. 39-44%) between different species, raising the possibility that species specificity could depend on specific features of chemical structure. Compounds were clustered by their chemical similarity, and similar compounds were examined for the expected similarity of their species-dependent liver effect profiles. In most cases, similar profiles were observed for members of the same cluster, but some compounds appeared as outliers. The outliers were the subject of focused assertion regeneration from MEDLINE as well as other data sources. In some cases, additional biological assertions were identified, which were in line with expectations based on compounds' chemical similarities. The assertions were further converted to binary annotations of underlying chemicals (i.e., liver effect vs no liver effect), and binary quantitative structure-activity relationship (QSAR) models were generated to predict whether a compound would be expected to produce liver effects in humans. Despite the apparent heterogeneity of data, models have shown good predictive power assessed by external 5-fold cross-validation procedures. The external predictive power of binary QSAR models was further confirmed by their application to compounds that were retrieved or studied after the model was developed. To the best of our knowledge, this is the first study for chemical toxicity prediction that applied QSAR modeling and other cheminformatics techniques to observational data generated by the means of automated text mining with limited manual curation, opening up new opportunities for generating and modeling chemical toxicology data. PMID- 20014753 TI - Conformational and solvation studies via computer simulation of the novel large scale diastereoselectively synthesized phosphinic MMP inhibitor RXP03 diluted in selected solvents. AB - Structure-activity relationship studies, regarding the influence of side chains of phosphinic pseudotripeptidic inhibitors on matrix metalloproteinases (MMPs), provided potent and selective inhibitors for this family of structurally and functionally related proteases. Among them, phosphinic pseudopeptide CbzPhepsi[P(O)(OH)CH(2)] phenylpropyl TrpNH(2), known as RXP03, has been extensively used for in vivo and in vitro studies so far. The large quantities of RXP03 required for in vivo studies, as well as the necessity for diastereoisomeric purity, motivated us to further explore and develop an efficient synthetic methodology, which allows separation of the four diastereoisomers of RXP03 based on the astonishing observed differences in solubility of the four isomers in various solvents. This fact prompted us to examine theoretically the conformational differences of these four isomers via computer simulations in the solvents used experimentally. Given the fact that the four examined diastereoisomeric forms of the phosphinic peptides exhibit different behavior in terms of potency and selectivity profiles toward zinc metalloproteases, this theoretical study provides valuable information on the conformation of phosphinic inhibitors and therefore improves the design and synthesis of active structures. The differences in solubility of RXP03 diastereoisomers in the used solvents were examined in terms of intra- and intermolecular structure. It is found that the different solubility of the RRS and RSS diastereoisomers in EtOH is a result of the different number of hydrogen bonds formed by each isomer with EtOH molecules. In the case of SRS and SSS in Et(2)O, their different solubility might be attributed to the different intramolecular hydrogen bonds formed on these diastereoisomers. PMID- 20014754 TI - Higher susceptibility to halothane modulation in open- than in closed-channel alpha4beta2 nAChR revealed by molecular dynamics simulations. AB - The neuronal alpha4beta2 nicotinic acetylcholine receptor (nAChR) is a potential molecular target for general anesthetics. It is unclear, however, whether anesthetic action produces the same effect on the open and closed channels. Computations parallel to our previous open channel study (J. Phys. Chem. B 2009, 113, 12581) were performed on the closed-channel alpha4beta2 nAChR to investigate the conformation-dependent anesthetic effects on channel structures and dynamics. Flexible ligand docking and over 20 ns molecular dynamics simulations revealed similar halothane-binding sites in the closed and open channels. The sites with relatively high binding affinities (approximately -6.0 kcal/mol) were identified at the interface of extracellular (EC) and transmembrane (TM) domains or at the interface between alpha4 and beta2 subunits. Despite similar sites for halothane binding, the closed-channel conformation showed much less sensitivity than the open channel to the structural and dynamical perturbations from halothane. Compared to the systems without anesthetics, the amount of water inside the pore decreased by 22% in the presence of halothane in the open channel but only by 6% in the closed channel. Comparison of the nonbonded interactions at the EC/TM interfaces suggested that the beta2 subunits were more prone than the alpha4 subunits to halothane binding. In addition, our data support the notion that halothane exerts its effect by disturbing the quaternary structure and dynamics of the channel. The study concludes that sensitivity and global dynamics responsiveness of alpha4beta2 nAChR to halothane are conformation dependent. The effect of halothane on the global dynamics of the open-channel conformation might also account for the action of other inhaled general anesthetics. PMID- 20014755 TI - Molecular modeling of two distinct triangular oligomers in amyloid beta-protein. AB - Amyloid-beta (Abeta) peptides exhibit many distinct structural morphology at the early aggregate stage, some of which are biological relevant to the pathogenesis of Alzheimer's disease (AD). Atomic-resolution structures of the early Abeta aggregates and their conformational changes in amyloid aggregation remain elusive. Here, we perform all-atom molecular modeling and dynamics simulations to obtain two stable triangular-like Abeta structures with the lowest packing energy, one corresponding to the Tycko's model (Paravastu, A.; Leapman, R.; Yau, W.; Tycko, R. Proc. Nat. Acad. Soc. U.S.A. 2008, 105, 18349-18354) (referred to C WT model) and the other corresponding to computational model (N-WT model). Both models have the same 3-fold symmetry but distinct beta-sheet organizations in which three Abeta hexamers pack together via either C-terminal beta-strand residues or N-terminal beta-strand residues forming distinct hydrophobic cross section. Structural and energetic comparisons of two 3-fold Abeta oligomers, coupled with structural changes upon the mutations occurring at the interacting interfaces, reveal that although hydrophobic interactions are still dominant forces, electrostatic interactions are more favorable in the N-WT model due to the formation of more and stable intersheet salt bridges, while solvation energy is more favorable in the C-WT model due to more exposed hydrophilic residues to solvent. Both models display many common features similar to other amyloid oligomers and therefore are likely to be biologically relevant. PMID- 20014756 TI - Compound I reactivity defines alkene oxidation selectivity in cytochrome P450cam. AB - Prediction of the chemoselectivity of drug oxidation by the human cytochrome P450 enzymes will aid in the avoidance of adverse drug reactions. The chemoselectivity of alkene oxidation is an important problem to address, as it can result in the formation of epoxides, which can have toxic effects. In this paper the epoxidation and hydroxylation of cyclohexene and propene by the bacterial P450(cam) isoform are modeled with hybrid quantum mechanical/molecular mechanical (QM/MM) methods. Snapshots for QM/MM modeling are chosen from molecular dynamics trajectories, to sample the different conformations of the enzyme-substrate complex. The energy barriers obtained for these processes are in qualitative agreement with experimental work, supporting the use of QM/MM methods in the study of selectivity for this class of enzyme. This work highlights the complexity involved in modeling these systems with QM/MM and the importance in the selection of starting geometries. PMID- 20014757 TI - Entrapment of individual DNA molecules and nanoparticles in porous alumina membranes. AB - Depth-resolved fluorescence imaging allows the motion of single DNA molecules and single nanoparticles at the liquid/solid interface to be recorded in real time. Porous alumina membranes were employed as model chromatographic packing material. Using a suitable pH and ionic strength, adsorptive interactions are suppressed. The effects of 3-dimensional topography, specifically the presence of nanopores, on DNA and nanoparticle migration across the surface are, thus, revealed. The residence times and the number of immobilized DNA molecules or particles increased as the pores size increased. Yet, we found that the pore diameter must be significantly larger than the particle diameter or the DNA short radius before entrapment can occur. Furthermore, the depth distribution of particles does not conform to one-dimensional diffusion in the pores, probably because of collisions with the walls. These observations provide new insights into conventional liquid chromatography as well as size-exclusion chromatography and membrane separations. PMID- 20014758 TI - Reaction of the black tea pigment theaflavin during enzymatic oxidation of tea catechins. AB - Degradation of the black tea pigment theaflavin was examined in detail. Enzymatic oxidation of a mixture of epigallocatechin and epicatechin initially produced theaflavin, while prolonged reaction decreased the product. Addition of ethanol to the reaction mixture at the point when theaflavin began to decrease afforded four new products, together with theanaphthoquinone, a known oxidation product of theaflavin. The structures of the new products were determined by spectroscopic methods. One of the products was an ethanol adduct of a theanaphthoquinone precursor, and this reacted with theaflavin to give two further products. A product generated by coupling of theaflavin with epicatechin quinone was also obtained. The structures of the products indicate that oxidation and coupling with quinones are key reactions in the degradation of theaflavins. The degradation of theaflavin probably contributes to production of thearubigins. PMID- 20014759 TI - Theoretical Raman optical activity study of the beta domain of rat metallothionein. AB - We present the calculated vibrational Raman optical activity (ROA) spectrum of the beta domain of rat metallothionein, which is the by far largest molecule for which full ab initio ROA calculations have been performed up to date with more than 400 atoms. While ROA signatures of regular secondary structure elements like beta-sheets and alpha-helices can be conveniently studied in terms of small model structures, this is no longer possible for more irregular proteins like metallothionein. The only secondary structure elements occurring in the molecule are turns, in particular beta turns. Our calculations reveal that especially bands in the wavenumber range from about 1100 to 1400 cm(-1) may be employed as signatures of such beta turns. This is also found in comparison to experimental data. In addition, good agreement between calculated and experimental spectra is found. PMID- 20014760 TI - Effects of fruit ellagitannin extracts, ellagic acid, and their colonic metabolite, urolithin A, on Wnt signaling. AB - Recent data suggest that ellagitannins (ETs), a class of hydrolyzable tannins found in some fruits and nuts, may have beneficial effects against colon cancer. In the stomach and gut, ETs hydrolyze to release ellagic acid (EA) and are converted by gut microbiota to urolithin A (UA; 3,8-dihydroxy-6H-dibenzopyran-6 one) type metabolites, which may persist in the colon through enterohepatic circulation. However, little is known about the mechanisms of action of either the native compounds or their metabolites on colon carcinogenesis. Components of Wnt signaling pathways are known to play a pivotal role in human colon carcinogenesis, and inappropriate activation of the signaling cascade is observed in 90% of colorectal cancers. This study investigated the effects of UA, EA, and ET-rich fruit extracts on Wnt signaling in a human 293T cell line using a luciferase reporter of canonical Wnt pathway-mediated transcriptional activation. The ET extracts were obtained from strawberry (Fragaria annassa), Jamun berry (Eugenia jambolana), and pomegranate (Punica granatum) fruit and were all standardized to phenolic content (as gallic acid equivalents, GAEs, by the Folin Ciocalteu method) and to EA content (by high-performance liquid chromatography methods): strawberry = 20.5% GAE, 5.0% EA; Jamun berry = 20.5% GAE, 4.2% EA; pomegranate = 55% GAE, 3.5% EA. The ET extracts (IC(50) = 28.0-30.0 microg/mL), EA (IC(50) = 19.0 microg/mL; 63 microM), and UA (IC(50) = 9.0 microg/mL; 39 microM) inhibited Wnt signaling, suggesting that ET-rich foods have potential against colon carcinogenesis and that urolithins are relevant bioactive constituents in the colon. PMID- 20014761 TI - Synthesis, crystal structure, and biological activity of 4-methyl-1,2,3 thiadiazole-containing 1,2,4-triazolo[3,4-b][1,3,4]thiadiazoles. AB - Heterocyclic compounds play an important role as the main sources of lead molecules of agrochemicals. Synthesis and biological activity of thiadiazole containing 1,2,4-triazolo[3,4-b][1,3,4]-thiadiazoles were seldom reported. To find novel lead compounds with various biological activities, a series of 6 substituted-3-(4-methyl-1,2,3-thiadiazolyl)[1,2,4]triazolo[3,4 b][1,3,4]thiadizoles were rationally designed and synthesized according to the principle of combinations of bioactive substructures by the condensation of 3-(4 methyl-1,2,3-thiadiazolyl)-4-amino-1,2,4-triazole-5-thione with various carboxylic acids and phosphorus oxychloride. All newly synthesized compounds were identified by proton nuclear magnetic resonance ((1)H NMR), infrared spectroscopy (IR), electroionization mass spectrometry (EI/MS), and elementary analysis. The crystal structure of 3-(4-methyl-1,2,3-thiadiazolyl)-6-(4 methylphenyl)[1,2,4]triazolo[3,4-b][1,3,4]thiadizole was determined by X-ray diffraction crystallography. In this crystal, two intermolecular hydrogen bonds (N2...H-C12 and N3...H-C13), a weak intermolecular interaction (S...S), and the weak ppi-ppi intermolecular interaction were observed. Fungicide screening indicated that all of the target compounds showed certain extent of growth inhibition against fungi tested. 3-(4-Methyl-1,2,3-thiadiazolyl)-6-n propyl[1,2,4]triazolo[3,4-b][1,3,4]thiadizole and 3-(4-methyl-1,2,3-thiadiazolyl) 6-trichloromethyl[1,2,4]triazolo[3,4-b][1,3,4]thiadizole were found to have potential wide spectrum of fungicide activity. The median effective concentrations (EC(50)) detected for 3-(4-methyl-1,2,3-thiadiazolyl)-6 trichloromethyl[1,2,4]triazolo[3,4-b][1,3,4]thiadizole to six fungi were from 7.28 micromol/L against Pellicularia sasakii (Shirai) to 42.49 micromol/L against Alternaria solani . The results indicated that thiadiazole-containing 1,2,4 triazolo[3,4-b][1,3,4]-thiadiazoles were potential fungicide lead compounds. PMID- 20014762 TI - Combined strategy for phytotoxicity enhancement of benzoxazinones. AB - Fifteen new derivatives of D-DIBOA, including aromatic ring modifications and the addition of side chains in positions C-2 and N-4, were synthesized and their phytotoxicity, selectivity, and structure-activity relationships evaluated. The most active compounds among the derivatives at the C-2 position were 6-Cl-2-Et-D DIBOA and 6-F-2-Et-D-DIBOA. Of the derivatives at N-4, the most active compounds were 6-Cl-4-Pr-D-DIBOA and 6-Cl-4-Val-D-DIBOA. These four compounds showed high levels of inhibition in root length at very low concentrations in all species. The most remarkable result is the 70% inhibition observed for the root length of cress at 100 nM caused by the latter two compounds. These results support our previous research and conclusions regarding the steric, electronic, and solubility requirements to achieve the maximum phytotoxic activity. PMID- 20014763 TI - Synthesis and insecticidal activity of heptafluoroisopropyl-containing benzoylphenylurea structures. AB - Fourteen novel heptafluoroisopropyl-containing benzoylphenylureas were designed and synthesized. Their insecticidal activities against armyworm ( Pseudaletia separata Walker) were examined and compared with the commercial product diflubenzuron. Three compounds (IIi, IIj, and IIk) showed excellent insecticidal effect, and their activity resembled that of diflubenzuron. Compound IIi also showed nearly the same insecticidal activity as novaluron on African cotton leafworm ( Spodoptera littoralis ). Furthermore, results from a field trial indicated that 5% EC IIi exhibited similar efficacy in comparison with chlorfluazuron and hexaflumuron against imported cabbage worm ( Pieris rapae L.) and diamondback moth ( Plutella xylostella ), respectively. PMID- 20014764 TI - Comparable susceptibilities of human 293 cells and insect Tn-5B1-4 cells to photoactivated alpha-terthienyl. AB - The hope is that photoactive compounds acting as potential insecticides will have reduced environmental risk, but that is not necessarily the case. In an attempt to elucidate the risk by which photoactivated alpha-terthienyl (alpha-T) affects human health, the effects of exposure of human 293 cells and insect Tn-5B1-4 cells to photoactivated alpha-T at certain doses were characterized. Photoactivated alpha-T exhibited dose dependence of toxicity and time kinetics of phototoxic activation on the growth of 293 cells (EC(50) = 6.23 microg/mL) and Tn 5B1-4 cells (EC(50) = 3.36 microg/mL). 293 cells appeared to be anchorage independent, inflated, and broken; Tn-5B1-4 cells showed significant necrosis. ROS productions and lipid peroxidation of 293 cells were always lower than that of Tn-5B1-4 cells in the treatments of alpha-T at the same dose. Moreover, photoactivated alpha-T caused nonselective DNA damage in 293 and Tn-5B1-4 cells at a 10 microg/mL dose and induced cell-cycle progression of 293 cells to increase apoptosis of cells and G1 arrest and decrease in S phase cell population, whereas Tn-5B1-4 cells showed S arrest accompanied by a dose dependent decrease in G1 and G2 phase cells at a 5 microg/mL dose. These observations suggest that Tn-5B1-4 cells are more susceptible to the action of photoactivated alpha-T than 293 cells, but photoactivated alpha-T as an efficient insecticide might be a potential factor in human mutagenic progression. PMID- 20014765 TI - Synthesis and biological activity of organotin 4-methyl-1,2,3-thiadiazole-5 carboxylates and benzo[1,2,3]thiadiazole-7-carboxylates. AB - A series of organotin 4-methyl-1,2,3-thiadiazole-5-carboxylates and benzo[1,2,3]thiadiazole-7-carboxylates have been synthesized and characterized by NMR ((1)H, (13)C, and (119)Sn), IR, and elemental analyses. The structure of the dimeric complex {[(BTHCO(2))SnEt(2)](2)O}(2) (BTH represents benzo[1,2,3]thiadiazol-7-yl) has been further confirmed by X-ray diffraction crystallography. Assessment for fungicidal activity indicates that all of the newly synthesized compounds exhibit good growth inhibition against Alternaria solani , Cercospora arachidicola , Gibberella zeae , Physalospora piricola , and Botrytis cinerea . High growth inhibition percentage at 50 microg/mL was obtained in vitro in the case of triorganotin 4-methyl-1,2,3-thiadiazole-5-carboxylates and benzo[1,2,3]thiadiazole-7-carboxylates. The corresponding EC(50) values of these triorganotin carboxylates have been detected, and values of EC(50) as low as 0.12 microg/mL against P. piricola and 0.16 microg/mL against G. zeae, respectively, were observed for triethyltin benzo[1,2,3]thiadiazole-7 carboxylate. PMID- 20014766 TI - Use of accurate mass full scan mass spectrometry for the analysis of anthocyanins in berries and berry-fed tissues. AB - Anthocyanins in extracts from raspberries and blueberries were analyzed by reversed-phase HPLC coupled to a high-resolution Exactive Orbitrap mass spectrometer (HR-MS) with a resolution of 100,000, operated with an electrospray source in the positive ionization mode. As consumption of anthocyanin-rich berry extracts has been associated with improved cognitive function, brain extracts from European greenfinches ( Carduelis chloris ) that had been fed one blackberry daily for a period of 2 weeks were analyzed by both HPLC with traditional tandem MS in the selected reaction monitoring mode and HPLC-HR-MS. Cyanidin-3-O glucoside was detected in the brain extracts by both methods, but because of its high level of selectivity, HR-MS was ca. 200-fold more sensitive. A further advantage of HR-MS is that unlike MS-SRM it enables both targeted and nontargeted compounds to be detected and much lower limits of detection are achieved without compromising the selectivity of the analysis. PMID- 20014767 TI - Reactivity of a disilylene [{PhC(NBu(t))(2)}Si](2) toward bromine: synthesis and characterization of a stable monomeric bromosilylene. AB - The reaction of the disilylene [{PhC(NBu(t))(2)}Si](2) (1) with 1 equiv of bromine in toluene afforded novel monomeric bromosilylene [{PhC(NBu(t))(2)}SiBr] (2). The result shows that the Si(I)-Si(I) bond in 1 was cleaved by bromine. An X ray structure of compound 2 has been determined. PMID- 20014768 TI - Diorganotelluride-catalyzed oxidation of silanes to silanols under atmospheric oxygen. AB - Diorganotellurides efficiently catalyze the aerobic oxidation of organosilanes under photosensitized conditions to afford the corresponding silanols in excellent yield. PMID- 20014769 TI - Silane-controlled diastereoselectivity in the tris(pentafluorophenyl)borane catalyzed reduction of alpha-diketones to silyl-protected 1,2-Diols. AB - B(C(6)F(5))(3)-catalyzed bis(hydrosilylation) of alpha-diketones can give high diastereomeric excess of either meso/anti (small silanes and disilane reagents) or dl/syn (bulky silanes) silyl-protected 1,2-diols. This easily tuned diastereoselectivity is rationalized based on the classic Felkin-Anh model applied to a mechanism relying on Si-H abstraction by the electrophilic borane reagent. PMID- 20014770 TI - Electron transfer photoredox catalysis: intramolecular radical addition to indoles and pyrroles. AB - The utilization of the photoredox catalyst, tris(2,2'-bipyridyl)ruthenium dichloride, and a household light bulb to effect radical cyclizations onto indoles and pyrroles at room temperature is reported. A reactive free radical intermediate is generated via the reduction of an activated C-Br bond by the single electron reductant, Ru(I), generated in a visible light induced photocatalytic cycle. This system represents an expansion of the application of photoredox catalysis in conventional free radical processes. PMID- 20014771 TI - DBU catalysis of N,N'-carbonyldiimidazole-mediated amidations. AB - 1,8-Diazabicyclo[5.4.0]undec-7-ene (DBU) has been found to catalyze the amidation of acyl imidazoles. The rate acceleration is especially evident with traditionally unreactive, electron-deficient anilines. DBU is readily available and offers safety and cost advantages over more commonly employed catalysts such as 1-hydroxybenzotriazole. PMID- 20014772 TI - Intrinsic propensities of amino acid residues in GxG peptides inferred from amide I' band profiles and NMR scalar coupling constants. AB - A reliable intrinsic propensity scale of amino acid residues is indispensable for an assessment of how local conformational distributions in the unfolded state can affect the folding of peptides and proteins. Short host-guest peptides, such as GxG tripeptides, are suitable tools for probing such propensities. To explore the conformational distributions sampled by the central amino acid residue in these motifs, we combined vibrational (IR, Raman, and VCD) with NMR spectroscopy. The data were analyzed in terms of a superposition of two-dimensional Gaussian distribution functions in the Ramachandran space pertaining to subensembles of polyproline II, beta-strand, right- and left-handed helical, and gamma-turn-like conformations. The intrinsic propensities of eight amino acid residues (x = A, V, F, L, S, E, K, and M) in GxG peptides were determined as mole fractions of these subensembles. Our results show that alanine adopts primarily (approximately 80%) a PPII-like conformation, while valine and phenylalanine were found to sample PPII and beta-strand-like conformations equally. The centers of the respective beta-strand distributions generally do not coincide with canonical values of dihedral angles of residues in parallel or antiparallel beta-strands. In fact, the distributions for most residues found in the beta-region significantly overlap the PPII-region. A comparison with earlier reported results for trivaline reveals that the terminal valines increase the beta-strand propensity of the central valine residue even further. Of the remaining investigated amino acids, methionine preferred PPII the most (0.64), and E, S, L, and K exhibit moderate (0.56-0.45) PPII propensities. Residues V, F, S, E, and L sample, to a significant extent, a region between the canonical PPII and (antiparallel) beta strand conformations. This region coincides with the sampling reported for L and V using theoretical predictions (Tran et al. Biochemistry 2005, 44, 11369). The distributions of all investigated residues differ from coil library and computationally predicted distributions in that they do not exhibit a substantial sampling of helical conformations. We conclude that this sampling of helical conformations arises from the context dependence, for example, neighboring residues, in proteins and longer peptides, some of which is long-range. PMID- 20014773 TI - Engineering of poly(butylcyanoacrylate) nanoparticles for the enhancement of the antitumor activity of gemcitabine. PMID- 20014774 TI - The spectral signatures of Frenkel polarons in H- and J-aggregates. AB - Electronic excitations in small aggregates, thin films, and crystals of conjugated organic molecules play a fundamental role in the operation of a wide array of organic-based devices including solar cells, transistors, and light emitting diodes. Such excitations, or excitons, are generally spread out over several molecules: a balance between the delocalizing influence of resonant intermolecular coupling and the localizing influence of static and dynamic disorder determines the coherence range of the exciton. Because of the "soft" nature of organic materials, significant nuclear relaxation in the participating molecules also accompanies the electronic excitations. To properly understand energy or charge transport, one must treat intermolecular (excitonic) coupling, electron-vibrational coupling, and disorder on equal footing. In this Account, we review the key elements of a theoretical approach based on a multiparticle representation that describes electronic excitations in organic materials as vibronic excitations surrounded by a field of vibrational excitations. Such composite excitations are appropriately called Frenkel excitonic polarons. For many conjugated molecules, the bulk of the nuclear reorganization energy following electronic excitation arises from the elongation of a symmetric vinyl stretching mode with energy approximately 1400 cm(-1). To appreciate the impact of aggregation, we study how the vibronic progression of this mode, which dominates the isolated (solvated) molecule absorption and emission spectra, is distorted when molecules are close enough to interact with each other. As we demonstrate in this Account, the nature of the distortion provides a wealth of information about how the molecules are packed, the strength of the excitonic interactions between molecules, the number of molecules that are coherently coupled, and the nature of the disorder. We show that the aggregation-induced deviations from the Poissonian distribution of vibronic peak intensities take on two extremes identified with ideal H- and J-aggregates. The sign of the nearest neighbor electronic coupling, positive for H and negative for J, distinguishes the two basic aggregate forms. For several decades, researchers have known that H aggregates exhibit blue-shifted absorption spectra and are subradiant while J aggregates exhibit the opposite behavior (red-shifted absorption and superradiance). However, the exact inclusion of exciton-vibrational coupling reveals several more distinguishing traits between the two aggregate types: in H(J)-aggregates the ratio of the first two vibronic peak intensities in the absorption spectrum decreases (increases) with increasing excitonic coupling, while the ratio of the 0-0 to 0-1 emission intensities increases (decreases) with disorder and increases (decreases) with increasing temperature. These two extreme behaviors provide the framework for understanding absorption and emission in more complex morphologies, such as herringbone packing in oligo(phenylene vinylene)s, oligothiophenes and polyacene crystals, as well as the polymorphic packing arrangements observed in carotenoids. PMID- 20014775 TI - Molecular scale assessment of methylarsenic sorption on aluminum oxide. AB - Methylated forms of arsenic (As), monomethylarsenate (MMA) and dimethylarsenate (DMA), have historically been used as herbicides and pesticides. Because of their large application to agriculture fields and the toxicity of MMA and DMA, the sorption of methylated As to soil constituents requires investigation. MMA and DMA sorption on amorphous aluminum oxide (AAO) was investigated using both macroscopic batch sorption kinetics and molecular scale extended X-ray absorption fine structure (EXAFS) and Fourier transform infrared (FTIR) spectroscopic techniques. Sorption isotherm studies revealed sorption maxima of 0.183, 0.145, and 0.056 mmol As/mmol Al for arsenate (As(V)), MMA, and DMA, respectively. In the sorption kinetics studies, 100% of added As(V) was sorbed within 5 min, while 78% and 15% of added MMA and DMA were sorbed, respectively. Desorption experiments, using phosphate as a desorbing agent, resulted in 30% release of absorbed As(V), while 48% and 62% of absorbed MMA and DMA, respectively, were released. FTIR and EXAFS studies revealed that MMA and DMA formed mainly bidentate binuclear complexes with AAO. On the basis of these results, it is proposed that increasing methyl group substitution results in decreased As sorption and increased As desorption on AAO. PMID- 20014776 TI - Fractional study of alkali-soluble hemicelluloses obtained by graded ethanol precipitation from sugar cane bagasse. AB - The two hemicellulosic fractions were subsequentially extracted with 5% and 8% NaOH aqueous solution at a solid to liquid ratio of 1:25 (g mL(-1)) at 50 degrees C for 3 h from the water, 1 and 3% NaOH-treated sugar cane bagasse, and subfractionated into six preparations by a graded ethanol precipitation method at concentrations of 15%, 30% and 60% (v/v). Sugar composition and molecular weight analysis showed that, with an increasing concentration of ethanol, hemicellulosic subfractions with both higher Ara/Xyl ratios and higher molecular weights were obtained. In other words, with an increasing ethanol concentration from 15% to 60%, the Ara/Xyl ratios increased from 0.043 in H(1) to 0.088 in H(3) and from 0.040 in H(4) to 0.088 in H(6), and the weight-average molecular weights of hemicellulosic subfractions increased from 42 430 (H(1)) to 85 510 (H(3)) g mol( 1) and from 46 130 (H(4)) to 64 070 (H(6)) g mol(-1), respectively. The results obtained by the analysis of Fourier transform infrared, sugar composition, and (1)H and (13)C nuclear magnetic spectroscopy showed that the alkali-soluble hemicelluloses had a backbone of xylose residues with a beta-(1-->4)-linkage and were branched mainly through arabinofuranosyl units at C-2 and/or C-3 of the main chain, whereas the differences may occur in the distribution of branches along the xylan backbone. PMID- 20014777 TI - Antiviral potential and molecular insight into neuraminidase inhibiting diarylheptanoids from Alpinia katsumadai. AB - At present, neuraminidase (NA) inhibitors are the mainstay of pharmacological strategies to fight against global pandemic influenza. In the search for new antiviral drug leads from nature, the seed extract of Alpinia katsumadai has been phytochemically investigated. Among the six isolated constituents, four diarylheptanoids showed in vitro NA inhibitory activities in low micromolar ranges against human influenza virus A/PR/8/34 of subtype H1N1. The most promising constituent, katsumadain A (4; IC(50) = 1.05 +/- 0.42 microM), also inhibited the NA of four H1N1 swine influenza viruses, with IC(50) values between 0.9 and 1.64 muM, and showed antiviral effects in plaque reduction assays. Considering the flexible loop regions of NA, extensive molecular dynamics (MD) simulations were performed to study the putative binding mechanism of the T shaped diarylheptanoid 4. Docking results showed well-established interactions between the protein and the core of this novel NA-inhibiting natural scaffold, excellent surface complementarity to the simulated binding pocket, and concordance with experimentally derived SAR data. PMID- 20014778 TI - Nonideal mixed micelles of fluorinated and hydrogenous surfactants in aqueous solution. NMR and SANS studies of anionic and nonionic systems. AB - Contrast variation SANS and (19)F chemical shifts were measured for three mixed equimolar micelle systems: sodium perfluorooctanoate (SPFO) and sodiumdecylsulfate (SDeS) in 200 mM NaCl, lithium perfluorononanate (LiPFN) and lithium dodecylsulfate (LiDS) in 200 mM LiCl, and a nonionic system C(8)F(17)C(2)H(4)(OC(2)H(4))(9) and C(12)H(25)(OC(2)H(4))(8) in water, all at 25 degrees C. The chemical shift measurements allow the calculation of the average fraction of nearest neighbors of each kind around the reporter group (the trifluoromethyl group). A preference for like neighbors were found in all systems, smallest in the SDeS/SPFO system and largest in the nonionic system, but in all cases substantially smaller than expected at critical conditions. From the SANS measurements the width of the micelle composition distribution was obtained. For the ionic systems similar values were obtained, showing a broadening compared to ideal mixtures, but not broad enough for demixing or clearly bimodal distributions. In the nonionic system the width was estimated as sigma = 0.18 and 0.22 using two different evaluation methods. These values suggest that the system is close to critical conditions. The lower value refers to a direct modeling of the system, assuming an ellipsoidal shape and a Gaussian composition distribution. The modeling showed the nonionic mixed micelles to be prolate ellipsoids with axial ratio 2.2 and an aggregation number larger than 100, whereas the two ionic systems fitted best to oblate shapes (axial ratios 0.8 and 0.65 for SDeS/SPFO and LiDS/LiPFN, respectively) and aggregation numbers of 60 for both. PMID- 20014779 TI - Concise total syntheses of the Lycopodium alkaloids (+/-)-nankakurines A and B via luciduline. AB - Total syntheses of the Lycopodium alkaloids nankakurines A and B have been accomplished in 6 and 7 steps, respectively, via a sequence that passes through a third Lycopodium alkaloid, luciduline, and forgoes the use of protecting groups on nitrogen. Key features include a short preparation of luciduline followed by a concise and stereoselective aminoallylation/ring-closing metathesis protocol to fashion the spiropiperidine ring common to nankakurines A and B. PMID- 20014780 TI - Highly sensitive matrix-assisted laser desorption ionization-mass spectrometry for high-throughput metabolic profiling. AB - In the present study, a high-throughput and nontargeted metabolomic technique using matrix-assisted laser desorption ionization-mass spectrometry (MALDI-MS) was developed for the rapid analysis of cellular metabolites. Either the detection limit or the linearity between concentrations of several standards and peak intensities was examined, indicating a detection limit lower than 10 fmol/well with a high linearity at low concentrations. To verify the validity of this method, metabolites from human acute lymphoblastic leukemia Jurkat cells were analyzed. Only 2,500 cells suspended in PBS were directly dropped onto a stainless MALDI sample plate, followed by mixing with matrix on the sample plate. Up to 150 metabolite peaks were detected from a single analysis within 90 s. For multivariate analysis of Jurkat cells against drug-treatment, three anticancer drugs were utilized. Principal component analysis of metabolites showed clear independent clusters for cells treated with these anticancer drugs. Furthermore, several metabolites involved in nucleotide synthesis were found to contribute to the separation of each cluster. These data suggest that the high-throughput MALDI MS-based metabolomic technique proposed in the present study can be utilized for drug screening and validation of drug efficacy and safety. PMID- 20014781 TI - Direct arylations of 2H-indazoles on water. AB - The efficient palladium-catalyzed synthesis of a range of substituted 2H Indazoles via C-H arylation is reported. Reactions are performed on water and provide a direct and mild route toward 2,3-diaryl indazoles of widespread biological significance. PMID- 20014782 TI - Total synthesis of the anti-apoptotic agents iso- and bongkrekic acids. AB - The first convergent total synthesis of isobongkrekic acid is reported involving three different stereospecific palladium cross-couplings for the formation of the diene units. Access to bongkrekic acid by this route is also demonstrated. These syntheses involve the formation of several potentially general building blocks. PMID- 20014783 TI - Cyclotetrasiloxanetetrols with methyl groups at silicon: isomers all-cis- and cis trans-cis-[MeSi(O)OH](4). AB - 1,3,5,7-Tetramethyl-1,3,5,7-tetrahydroxycyclotetrasiloxane has been synthesized in good yield from tetrapotassium tetramethylcyclotetrasiloxanolate. Methods for isolation of all-cis-isomer I and cis-trans-cis-isomer II as the only crystalline product have been developed. Both isomers have been characterized by single crystal X-ray diffraction analysis (XRD), elemental analysis, NMR ((1)H, (13)C, (29)Si), and IR. The stability of these compounds in solutions and the solid state has been investigated. PMID- 20014784 TI - Accurate calculated optical properties of substituted quaterphenylene nanofibers. AB - The accurate prediction of both excitation and emission energies of substituted p quaterphenylenes using a variety of established and newly developed density functional methods is evaluated and compared against experimental data, both from single molecules and from nanofibers. For calculation of the UV-vis excitation the MPW1K functional is the best performing method (with the employed TZVP basis set). After a linear scaling factor is applied, mPW2-PLYP, CIS and the very fast INDO/S also reproduce the experimental data correctly. For the fluorescence relaxation energies MPW1K, mPW2-PLYP, and INDO/S give good results, even without scaling. However, mPW2-PLYP involves second-order perturbation to introduce nonlocal electron correlation and therefore requires significantly more resources, so the recommended level of theory for a single methodology to investigate the optical properties of substituted phenylenes and related systems is MPW1K/6-311+G(2d,p), followed by INDO/S as a low-cost alternative. As an extension of a previous work on predicting first hyperpolarisabilities, we can now demonstrate that the chosen approach (HF/6-31G(d)//B3LYP/6-31G(d)) produces data that correlate well with the susceptibilities derived from measurements on nanofibers. PMID- 20014785 TI - Small-angle neutron scattering studies of phospholipid-NSAID adducts. AB - Nonsteroidal anti-inflammatory drugs (NSAIDs) are known to have strong interactions with lipid membranes. Using small-angle neutron scattering, the effect of ibuprofen, a prominent NSAID, on the radius of small unilamellar vesicles of 1,2-dimyristoyl-sn-glycero-3-phosphatidylcholine (DMPC) and their bilayer structure was studied systematically as a function of pH (ranging from 2 to 8) and drug-to-lipid mole ratio (from 0/1 to 0.62/1 mol/mol). Ibuprofen with a pK(a) of approximately 4.6 was found to significantly affect the bilayer structure at all pH values, irrespective of the charge state of the drug. At low pH values, the drug reduces the bilayer thickness, induces fluid-like behavior, and changes headgroup hydration. The incorporation of the drug in the lipid bilayer while affecting the local bilayer structure and hydration of the lipid does not affect the overall stability of the vesicle dispersions over the pH range studied. PMID- 20014786 TI - L-proline-catalyzed three-component domino [3+2+1] annulation for the regio- and diastereoselective synthesis of highly substituted thienothiopyrans containing three or four stereocenters. AB - L-proline-catalyzed three-component reactions of ethyl 2-[(2-oxo-2 arylethyl)sulfonyl]acetate or ethyl 2-[(2-ethoxy-2-oxoethyl)sulfonyl]acetate, aromatic aldehydes, and 5-aryltetrahydro-3-thiophenone furnished a variety of highly substituted thieno[3,2-c]thiopyran derivatives. This facile transformation presumably occurs via a one-pot domino sequence of enamine formation/aldol condensation/Michael addition/6-exo-trig cyclization/elimination and involves the creation in a single operation of three C-C bonds and the generation of three new stereocenters with complete diastereoselectivity in all cases and a fourth one in ca. 7:3 diastereomeric ratio when starting from a 5-substituted tetrahydro-3 thiophenone derivative. PMID- 20014787 TI - Observations on the deprotection of pinanediol and pinacol boronate esters via fluorinated intermediates. AB - Methods for the deprotection of pinanediol and pinacol esters of various boronic acids via fluoroborane intermediates were evaluated. Treatment of the boronate esters with potassium hydrogen difluoride normally gives trifluoroborate salts; in the case of alpha-amido alkyl or o-amido phenyl boronate esters, aqueous workup gives difluoroboranes. Procedures for transformation of both trifluoroborates and difluoroboranes to free boronic acids are described. PMID- 20014788 TI - Crystal structures and properties of large protonated water clusters encapsulated by metal-organic frameworks. AB - A large ionic water cluster H(H(2)O)(28)(+), consisting of a water shell (H(2)O)(26) and an encaged species H(H(2)O)(2)(+) as a center core, was trapped in the well-modulated cavity of a porous metal-organic framework (MOF) {[Co(4)(dpdo)(12)(PMo(12)O(40))(3)](-)}(infinity) and structurally characterized. Degeneration of the protonated water cluster H(H(2)O)(28)(+) into a smaller cluster H(H(2)O)(21)(+) and recovery of H(H(2)O)(28)(+) from the resulting H(H(2)O)(21)(+) cluster in a reversible way demonstrated the unusual stability of the protonated water clusters H(H(2)O)(28)(+) and H(H(2)O)(21)(+) in the robust crystal host. Proton transport and proton/potassium ion exchange through the channels of the crystal host have been investigated by a well-established fluorometry method. X-ray fluorescence experiments and X-ray structural analyses of the exchanged crystals confirmed the occurrence of the proton/potassium ion exchange reaction and the transformation of the protonated water cluster H(H(2)O)(28)(+) to an ionic cluster K(H(2)O)(27)(+). Comparison of the H(+)/K(+) exchange of H(H(2)O)(28)(+) with that of its neighboring protonated water cluster H(H(2)O)(27)(+) suggested that the abundance of hydrogen bonds associated with the hydronium/water cluster in the H(H(2)O)(28)(+) cluster was essential for proton transport through the Grotthuss mechanism. On the basis of the results, our porous network could be described as a synthetic non-peptide ion channel, in terms of not only structural features but also the functions addressed. Direct observation of the structures of various large ionic water clusters trapped by porous MOFs, coupled with the proton/ion-exchange processes and the reversible dehydration/rehydration, provided valuable insights into the aqueous proton transfer and its mobility pertaining to the large protonated water clusters in the condensed phase. PMID- 20014789 TI - Free solution hydrodynamic separation of DNA fragments from 75 to 106,000 base pairs in a single run. AB - Gel electrophoresis is commonly used to separate DNA, but narrow capillaries or microchannels desired for high throughput efficient separations are difficult to fill with gels. We report here that a narrow capillary can be used to hydrodynamically separate a wide size range of DNA fragments in a single run without the need for gels, wall coatings, or an electric field. We also demonstrate that attractive separation is possible in a few minutes and that the separated DNA can be collected into individual fractions that remain viable for amplification via the polymerase chain reaction. PMID- 20014791 TI - A porous coordination network catalyzes an olefin isomerization reaction in the pore. AB - All-trans retinal efficiently diffused into the pore of a porous coordination network consisting of ZnI(2) and an electron-deficient triazine-cored ligand. Enclathrated retinal was isomerized into the 13-cis form and easily replaced with all-trans retinal in solution, thus revealing the catalysis of retinal isomerization by the porous network. PMID- 20014792 TI - Sonochemical formation of methyl hydroperoxide in polar aprotic solvents and its effect on single-walled carbon nanotube dispersion stability. AB - Ultrasonication is a common method for dispersing nanoparticles and colloids. We have found that, under certain conditions, unintended sonochemical reactions can be initiated by the incident ultrasonic energy, yielding unwanted byproducts. In this work, we determined that methyl hydroperoxide can be produced by an autoxidation chain reaction when ultrasonicating polar aprotic solvents containing methyl groups. Methyl radicals were detected during ultrasonication by their interaction with lucigenin, which emits sonochemiluminescence. A colorimetric triiodide test was used to confirm the presence of a hydroperoxide. The concentration of methyl hydroperoxide as a function of the ultrasonication time was measured by titration with NaOH. When above the critical coagulation concentration, this sonochemical byproduct collapses the electrical double layer, disrupting the dispersion stability and lowering the dispersion limits. This is significant when developing ultrasonication processes for dispersion of nanoparticles and colloids. There are no other examples of sonochemically initiated solvent autoxidation destabilizing single-walled carbon nanotube dispersions reported in the literature. PMID- 20014790 TI - Heme-coordinating inhibitors of neuronal nitric oxide synthase. Iron-thioether coordination is stabilized by hydrophobic contacts without increased inhibitor potency. AB - The heme-thioether ligand interaction often occurs between heme iron and native methionine ligands, but thioether-based heme-coordinating (type II) inhibitors are uncommon due to the difficulty in stabilizing the Fe-S bond. Here, a thioether-based inhibitor (3) of neuronal nitric oxide synthase (nNOS) was designed, and its binding was characterized by spectrophotometry and crystallography. A crystal structure of inhibitor 3 coordinated to heme iron was obtained, representing, to our knowledge, the first crystal structure of a thioether inhibitor complexed to any heme enzyme. A series of related potential inhibitors (4-8) also were evaluated. Compounds 4-8 were all found to be type I (non-heme-coordinating) inhibitors of ferric nNOS, but 4 and 6-8 were found to switch to type II upon heme reduction to the ferrous state, reflecting the higher affinity of thioethers for ferrous heme than for ferric heme. Contrary to what has been widely thought, thioether-heme ligation was found not to increase inhibitor potency, illustrating the intrinsic weakness of the thioether-ferric heme linkage. Subtle changes in the alkyl groups attached to the thioether sulfur caused drastic changes in the binding conformation, indicating that hydrophobic contacts play a crucial role in stabilizing the thioether-heme coordination. PMID- 20014793 TI - Delta12-prostaglandin J2 as a product and ligand of human serum albumin: formation of an unusual covalent adduct at His146. AB - Human serum albumin (HSA), the most abundant protein in plasma, has a very unique function, catalyzing the conversion of prostaglandin J(2) (PGJ(2)), a dehydration product of PGD(2), to yield Delta(12)-PGJ(2). These PGD(2) metabolites are actively transported into cells and accumulated in the nuclei, where they act as potent inducers of cell growth inhibition and cell differentiation, and exhibit their own unique spectrum of biological effects. The facts that (i) arachidonic acid metabolites bind to human serum albumin (HSA) and the metabolism of these molecules is altered as a result of binding, (ii) HSA catalyzes the transformation of PGJ(2) into Delta(12)-PGJ(2), and (iii) Delta(12)-PGJ(2) is stable in serum suggest that HSA may bind and stabilize Delta(12)-PGJ(2) in a specific manner. A molecular interaction analysis using surface plasmon resonance (Biacore) indeed suggested the presence of a specific Delta(12)-PGJ(2)-binding site in HSA. To investigate the molecular details of the binding of this PGD(2) metabolite to albumin, we analyzed the cocrystal structure of the HSA-Delta(12) PGJ(2)-myristate complex by X-ray crystallography and found that two Delta(12) PGJ(2) molecules bind to a primary site in subdomain IB of the protein. The electron density results suggested that one of the two Delta(12)-PGJ(2) molecules that specifically bind to the site covalently interacted with a histidine residue (His146). Using nano-LC-MS/MS analysis of the HSA-Delta(12)-PGJ(2) complex, the formation of an unusual Delta(12)-PGJ(2)-histidine adduct at His146 was confirmed. Thus, our crystallographic and mass spectrometric analyses of the HSA Delta(12)-PGJ(2) complex provided intriguing new insights into the molecular details of how this electrophilic ligand interacts with its primary producer and transporter. PMID- 20014795 TI - Crystal structure and magnetism of a well isolated 2D-quantum Heisenberg antiferromagnet, (Quinolinium)(2)CuBr(4).2H(2)O, and its anhydrous form. AB - Reaction of quinoline with HBr and CuBr(2) generates a mixture of two compounds, (quinolinium)(2)CuBr(4).2H(2)O (1) and (quinolinium)(2)CuBr(4) (2) for which single-crystal X-ray structures have been solved. Compound 1 crystallizes in the monoclinic space group C2/c as layers of tetrabromocuprate ions which are separated by intervening layers of quinolinium ions. Compound 2 crystallizes in the triclinic space group P1. Magnetic data analysis reveals that 1 behaves as a 2D-quantum Heisenberg antiferromagnet with 2J/k(B) = -6.17(3) K within the layers. High field magnetization data at low temperatures suggests that T(N) must be less than 1.8 K for 1, yielding a figure of merit |k(B)T(N)/2J| < 0.29, which indicates excellent isolation between the layers. Magnetic exchange in compound 2 was much weaker and was fit to a linear chain antiferromagnet with 2J/k(B) = 1.59(3) K. PMID- 20014796 TI - Synthesis and structural studies of heterobimetallic alkoxide complexes supported by bis(phenolate) ligands: efficient catalysts for ring-opening polymerization of L-lactide. AB - A series of heterobimetallic titanium(IV) complexes [LTi(O(i)Pr)(mu O(i)Pr)(2)Li(THF)(2)], [LTi(O(i)Pr)(mu-O(i)Pr)(2)Na(THF)(2)], [LTi(mu O(i)Pr)(2)Zn(O(i)Pr)(2)], and [LTi(mu-O(i)Pr)(2)Mg(O(i)Pr)(2)] (where L = bidentate bisphenol ligands) have been synthesized and characterized including a structural determination of [L(1)Ti(mu(2)-O(i)Pr)(2)(O(i)Pr)Li(THF)(2)] (1a). These complexes were investigated for their utility in the ring-opening polymerization (ROP) of l-lactide (LA). Polymerization activities have been shown to correlate with the electronic properties of the substituent within the bisphenol ligand. In contrast to monometallic titanium initiator 1e, all the heterobimetallic titanium initiators (Ti-Li, Ti-Na, Ti-Zn, and Ti-Mg) show enhanced catalytic activity toward ring-opening polymerization (ROP) of l-LA. In addition, the use of electron-donating methoxy or methylphenylsulfonyl functional ligands reveals the highest activity. The bisphenol bimetallic complexes give rise to controlled ring-opening polymerization, as shown by the linear relationship between the percentage conversion and the number-average molecular weight. The polymerization kinetics using 2c as an initiator were also studied, and the experimental results indicate that the reaction rate is first-order with respect to both monomer and catalyst concentration with a polymerization rate constant, k = 81.64 M(-1) min(-1). PMID- 20014797 TI - Structural mapping and framework interconversions in 1D, 2D, and 3D divalent metal R,S-hydroxyphosphonoacetate hybrids. AB - Reactions of divalent cations (Mg(2+), Co(2+), Ni(2+), and Zn(2+)) with R,S hydroxyphosphonoacetic acid (HPAA) in aqueous solutions (pH values ranging 1.0 4.0) yielded a range of crystalline hydrated M-HPAA hybrids. One-dimensional (1D) chain compounds were formed at room temperature whereas reactions conducted under hydrothermal conditions resulted in two-dimensional (2D) layered frameworks or, in some cases, three-dimensional (3D) networks incorporating various alkaline cations. 1D phases with compositions [M{HO(3)PCH(OH)CO(2)}(H(2)O)(2)].2H(2)O (M = Mg, Co, and Zn) were isolated. These compounds were dehydrated in liquid water to yield the corresponding [M{HO(3)PCH(OH)CO(2)}(H(2)O)(2)] compounds lacking the lattice water between the 1D chains. [M{HO(3)PCH(OH)CO(2)}(H(2)O)(2)] (M = Mg, Ni, Co, Zn) compounds were formed by crystallization at room temperature (at higher pH values) or also by partial dehydration of 1D compounds with higher hydration degrees. Complete dehydration of these 1D solids at 240-270 degrees C led to 3D phases, [M{HO3PCH(OH)CO(2)}]. The 2D layered compound [Mg{HO(3)PCH(OH)CO(2)}(H(2)O)(2)] was obtained under hydrothermal conditions. For both synthesis methods, addition of alkali metal hydroxides to adjust the pH usually led to mixed phase materials, whereas direct reactions between the metal oxides and the hydroxyphosphonoacetic acid gave single phase materials. On the other hand, adjusting the pH with acetate salts and increasing the ratio M(2+)/HPAA and/or the A(+)/M(2+) ratio (A = Na, K) resulted in 3D networks, where the alkali cations were incorporated within the frameworks for charge compensation. The crystal structures of eight new M(II)-HPAA hybrids are reported herein and the thermal behavior related to dehydration/rehydration of some compounds are studied in detail. PMID- 20014798 TI - Sigma-donor and pi-acceptor properties of phosphorus ligands: an insight from the natural orbitals for chemical valence. AB - The bonding between phosphorus ligands X = PCl(3), PF(3), P(OCH(3))(3), PH(3), PH(2)CH(3), PH(CH(3))(2), P(CH(3))(3) and the metal-containing fragments [Ni(CO)(3)], [Mo(CO)(5)], and [Fe(CO)(4)] have been studied by Natural Orbitals for Chemical Valence (NOCV). The main attention was paid to estimation of donor (Deltaq(d)) /acceptor (Deltaq(bd)) properties of X on the basis of NOCV's charge criterion. All ligands X are found to be both sigma-donors and pi-acceptors. The best sigma-donor and pi-acceptor ligands are P(CH(3))(3) and PY(3) (Y horizontal line F,Cl), respectively, in both the nickel and molybdenum complexes. The NOCV contributions to deformation density show that the sigma-component corresponds to the donation from the lone electron pair of phosphorus, enhanced further by a transfer from ancillary halogen atoms (in the case of PCl(3) and PF(3)) to a bonding region and to oxygen atoms of carbonyls. The pi-bonding is due to the electron transfer from the metal into the empty orbital of X, mostly exhibiting phosphorus 3p character. It was shown that within the molecular orbital framework, the trend for the donor/acceptor strength of X can be explained by the difference in the orbital energies of the orbitals involved in the donation/back donation. Regarding the influence of the metal fragment on the donor/acceptor properties of X, it was demonstrated that the relative order of the phosphorus ligands remains in general intact. The only exception is the P(OCH(3))(3) ligand changing its position in molybdenum series compared to the nickel complexes. However, a change in the metal-containing fragment can influence the magnitude of electron transfer. For the set of phosphorus ligands studied here the effect is much less pronounced than for other ligands studied previously. PMID- 20014799 TI - Origin of high photoluminescence efficiencies in CdSe quantum belts. AB - CdSe quantum belts (QBs) having lengths of 0.5-1.5 microm and thicknesses of 1.5 2.0 nm exhibit high photoluminescence (PL) efficiencies of approximately 30%. Epifluorescence studies establish the PL spectra to be uniform along single QBs, and nearly the same from QB to QB. Photogenerated excitons are shown to be effectively delocalized over the entire QBs by position-selective excitation. Decoration of the QBs with gold nanoparticles indicates a low density of surface trap sites, located primarily on the thin belt edges. The high PL efficiencies and effective exciton delocalization are attributed to the minimization of defective {1100} edge surface area or edge-top/bottom (face) line junctions in QBs relative to quantum wires having roughly isotropic cross sections, for which very low PL quantum efficiencies have been reported. The results suggest that trap sites can be minimized in pseudo-one-dimensional nanocrystals, such that the facile transport of energy and charge along their long axes becomes possible. PMID- 20014800 TI - Stolonoxides E and F, cytotoxic metabolites from the marine ascidian Stolonica socialis. AB - Two new members of the stolonoxide family, stolonoxides E (1) and F (2), were isolated from samples of the marine ascidian Stolonica socialis collected in Cadiz (Spain). Their structures were determined by a combination of techniques, including (+)-HRESIMS, 1D and 2D NMR spectroscopy, and comparison with published data for the structurally related stolonoxides A-D (3-6). Both compounds displayed cytotoxicity against a panel of three human tumor cell lines. PMID- 20014801 TI - Growth pattern of truncated octahedra in al(n) (N H(2)O + NCCCC(-)(OEt)(CN) --> (NCCCCCN)(* ) + EtO(*). A comparison of the neutralization/reionization ((-)NR(+)) and charge reversal ((-)CR(+)) spectra of (NCCCCCN)(*-) indicate that some neutrals NCCCCCN are energized and rearrange to an isomer which decomposes by loss of carbon. An ab initio study at the CCSD(T)/cc-pVTZ//B3LYP/6-311+G(3df) level of theory indicates that (i) triplet NCCCCCN is the ground state with a T/S energy gap of 14.9 kcal mol(-1); (ii) the structures of triplet and singlet NCCCCCN need to be described by molecular obital theory, and a simple valence bond approach cannot be used for this system; and (iii) there are several possible routes by which an energized neutral may lose carbon, but the major route involves the triplet nitrile to isonitrile rearrangement NCCCCCN --> CNCCCCN --> NCCCCN + C. PMID- 20014807 TI - Kinematic slowing and electrostatic guiding of KBr molecules formed by the reactive collision process: K + HBr --> KBr + H. AB - We have generated a beam of translationally cold KBr molecules formed by exoergic reactive collisions in counterpropagating beams of K atoms and HBr molecules. The method relies on the extreme mass ratio of the products and the proper choice of the beam velocities (Liu, N.-N.; Loesch, H. J. Phys. Rev. Lett. 2007, 98, 10300). Here, we report the results of experiments on guiding the slow molecules from the site of their creation to the detector by a linear electrostatic quadrupole field. The device enhances the total intensity by a factor of 2.3 and the intensity at 14.2 m/s (1.4K) by an order of magnitude. The density velocity distributions, deduced directly from the observed time-of-flight profiles, peak at 20 m/s (2.9K). A numerical simulation of the guiding efficiency indicates that the polarization of the nascent molecules is first altered by a sudden change of the quantization axis from parallel to the initial relative velocity to parallel to the fringing field and thereafter follows adiabatically the local field as quantization axis. Drastic differences between the velocity and rotational state distributions of the molecules entering and leaving the energized quadrupole field are predicted. The counterpropagating beams can be used to continuously load an electrostatic trap. The equilibrium density of confined molecules is estimated to 1 x 10(7) cm(-3). PMID- 20014809 TI - Infrared spectra of CH(2)=Zr(H)NC, CH(3)-ZrNC, and eta(2)-Zr(NC)-CH(3) produced by reactions of laser-ablated Zr atoms with acetonitrile. AB - The zirconium methylidene isocyanide, methyl isocyanide, and eta(2)-nitrile-pi complexes are observed in the matrix IR spectra from reactions of laser-ablated Zr atoms and acetonitrile isotopomers. The methylidene CH(2)=Zr(H)NC has a C(1) agostic structure in line with simple early transition-metal methylidenes recently produced from reactions with small alkanes and methyl halides, and the extent of agostic distortion is also comparable. Formation of the isocyanide complexes from acetonitrile is interesting but not surprising according to previous studies of metal reactions with nitrile-containing compounds, and their stabilities over the cyanide species are reproduced by DFT calculations. Observation of the relatively rare nitrile pi-complex and its photodissociation suggests that the reaction proceeds in the order of Zr<--NCCH(3), eta(2)-Zr(NC) CH(3), CH(3)-ZrNC, and CH(2)=Zr(H)NC. The intermediate transition-state structures are also examined. PMID- 20014805 TI - Efficient formation of the tandem thymine glycol/8-oxo-7,8-dihydroguanine lesion in isolated DNA and the mutagenic and cytotoxic properties of the tandem lesions in Escherichia coli cells. AB - Reactive oxygen species can induce the formation of not only single-nucleobase lesions, which have been extensively studied, but also tandem lesions. Herein, we report a high frequency of formation of a type of tandem lesion, where two commonly observed oxidatively induced single-nucleobase lesions, that is, thymidine glycol (Tg) and 8-oxo-7,8-dihydro-2'-deoxyguanosine (8-oxodG), are vicinal to each other in calf thymus DNA upon exposure to Cu(II)/ascorbate along with H(2)O(2) or gamma-rays. We further explored how the tandem lesions perturb the efficiency and fidelity of DNA replication by assessing the replication products formed from the propagation, in Escherichia coli cells, of the single stranded pYMV1 shuttle vectors containing two tandem lesions [5'-(8-oxodG)-Tg-3' and 5'-Tg-(8-oxodG)-3'] or an isolated Tg or 8-oxodG. The bypass efficiencies for the two tandem lesions were approximately one-half of those for the two isolated single-nucleobase lesions. The presence of an adjacent Tg could lead to significant increases in G-->T transversion at the 8-oxodG site as compared to that of a single 8-oxodG lesion; the frequencies of G-->T mutation were approximately 18, 32, and 28% for 8-oxodG that is isolated, in 5'-(8-oxodG)-Tg-3' and in 5'-Tg-(8-oxodG)-3', respectively. Moreover, both pol IV and pol V are involved, in part, in bypassing the Tg, either present alone or as part of the tandem lesions, in E. coli cells. Together, our results support that complex lesions could exert greater cytotoxic and mutagenic effects than when the composing individual lesions are present alone. PMID- 20014810 TI - Rotationally resolved ground state vibrational levels of HC2S studied by two color resonant four-wave mixing. AB - A high-resolution study of the X(2)Pi(3/2) ground state rovibronic energy levels of jet-cooled HC(2)S radical using the double-resonance two-color four-wave mixing technique (TC-RFWM) is reported. The rotational structure of the bands is observed by excitation of selected R-branch lines in the origin band of the A(2)Pi(3/2)-X(2)Pi(3/2) electronic system. The second laser frequency is scanned to transfer the population from the rotational level(s) of the upper state to selected vibronic levels of the ground state. Fourteen rotationally resolved vibrational bands have been recorded for energies up to 1800 cm(-1) above the v'' = 0 X(2)Pi(3/2) electronic ground state. Effective rotational constants and origins are determined for levels that involve fundamental and overtone combinations of the nu(3) (CS stretch), nu(4) (CCH bend), and nu(5) (CCS bend) vibrations. This illustrates the power and advantages of the TC-RFWM approach for the study of the ground state manifold of reactive intermediates produced in low concentrations with high resolution, good signal-to-noise and wide dynamic range. PMID- 20014811 TI - Comment on "New interpretation of the CH stretching vibrations in imidazolium based ionic liquids". PMID- 20014813 TI - Helical C(2) structure of perfluoropentane and the C(2v) structure of perfluoropropane. AB - Saturated hydrocarbons have structures with completely staggered bonds and dihedral angles of 180 degrees . Substituting hydrogen by fluorine results in a slight shift from 180 degrees , giving rise to a helical structure. X-ray diffraction studies on fibers and computational studies on perfluoroalkanes estimate a dihedral angle of about 17 degrees from the trans position. The rotational spectra of perfluoropentane and its three (13)C isotopomers have been observed and assigned using a pulsed-jet Fourier transform microwave spectrometer. The rotational constants for the parent species are A 990.6394(3) MHz, B 314.0002(1) MHz, and C 304.3703(1) MHz, respectively. The determination of an exact dihedral angle has been challenging, as the helical twist has proven to be quite sensitive to the structural inputs and constraints. A series of r(0) structures incorporating various model constraints and a Kraitchman analysis gives a range of 13-19 degrees for the torsional angle. An objective approach, which only assumes overall C(2) symmetry, is to scale the principal coordinates from ab initio models by the square root of the ratio of the observed second moments to the computed second moments. The scaled structures of computed models at various levels of theory reproduce the parent second moments exactly and the (13)C second moments very well, giving a dihedral angle of 17 +/- 1 degrees from trans. The microwave spectrum of perfluoropropane has also been observed and assigned. The rotational constants are A 1678.5982(9) MHz, B 900.1968(10) MHz, and C 955.3216(11) MHz, respectively. Unlike longer perfluoroalkanes, perfluoropropane has a nonhelical, C(2v) structure. Computations are in excellent agreement with experimental results. PMID- 20014812 TI - Wavelength and time-resolved imaging of material ejection in infrared matrix assisted laser desorption. AB - The dynamics of glycerol ablation at atmospheric pressure was studied using fast photography. A mid-infrared optical parametric oscillator was used to irradiate a droplet of glycerol at normal incidence. The wavelength of the infrared source was tunable and was varied between 2.7 and 3.5 microm for the studies. After an adjustable delay, an excimer pumped dye laser was used to illuminate the expanding plume, and the 90 degrees scattered light was imaged with a high-speed CMOS camera. The time delay between the IR and UV lasers was varied up to 1 ms with a particular emphasis in the early stages of plume evolution below 1 micros. The threshold fluence for plume formation varied between 1000 and 6000 J/m(2), and the minimum fluence corresponded to the OH stretch absorption of glycerol near 3.0 microm, which also corresponded to the greatest scattered light signal and duration of material emission. The velocity of the expanding plume was measured and ranged from >300 m/s near the OH stretch absorption to <100 m/s near the 3.4 microm CH stretch. Plume modeling calculations suggest that material removal is driven by phase explosion in the stress confinement regime that is at a maximum near the wavelength of the OH stretch absorption. PMID- 20014814 TI - Theoretical investigation of N-nitrosodimethylamine formation from nitrosation of trimethylamine. AB - Tertiary amines have been demonstrated to be capable of undergoing nitrosative cleavage to produce carcinogenic N-nitrosamines. The reaction mechanism of the nitrosation of trimethylamine (TMA) to produce N-nitrosodimethylamine (NDMA) was investigated at the CBS-QB3 level of theory. The formation of NDMA from TMA was proposed to be initiated by the formation of an iminium ion, Me(2)N(+) horizontal lineCH(2). Two different mechanisms (NOH elimination mechanism and oxidation abstraction mechanism) leading to Me(2)N(+) horizontal lineCH(2) were investigated, and the oxidation abstraction mechanism was found to be mainly operative. This result indicates that the oxidation abstraction mechanism plays an important role in the nitrosation of both N,N-dialkyl aromatic and tertiary aliphatic amines. Starting from the iminium ion, two experimentally proposed mechanisms (pathways 1 and 2) and one new mechanism (pathway 3) were examined. Pathway 1 proposes that the iminium ion undergoes hydrolysis to give dimethylamine (DMA), which then can be further nitrosated to NDMA; pathway 2 proposes that the iminium ion reacts with NO(2)(-) and forms a neutral intermediate, which then collapses to NDMA. In pathway 3, the iminium ion reacts with N(2)O(3) to give NDMA. Calculation results indicate that in aqueous solution pathway 1 is more feasible than pathways 2 and 3; moreover, the transformation from the iminium ion to NDMA is the rate-determining step. This work will be helpful to elucidate the formation mechanisms of the carcinogenic N-nitrosamines from the nitrosation of tertiary amines. PMID- 20014815 TI - Spectroscopic and kinetic properties of HO(2) radicals and the enhancement of the HO(2) self reaction by CH(3)OH and H(2)O. AB - The line center absorption cross sections and the rate constants for self reaction of hydroperoxy radicals (HO(2)) have been examined in the temperature range of 253-323 K using pulsed laser photolysis combined with tunable diode laser absorption in the near-IR region. The transition probed was in the 2nu(1) OH overtone transition at 1506.43 nm. The temperature dependence of the rate constant (k) for the HO(2) + HO(2) reaction was measured relative to the recommended value at 296 K, giving k = (3.95 +/- 0.45) x 10(-13) x exp[(439 +/- 39)/T] cm(3) molecule(-1) s(-1) at a total pressure of 30 Torr (N(2) + O(2)). After normalizing our determination and previous studies at low pressure, we recommend k = (2.45 +/- 0.50) x 10(-13) x exp[(565 +/- 130)/T] cm(3) molecule(-1) s(-1) (0 < P < 30 Torr, 95% confidence limits). The observed rate coefficient, k(obs), increases linearly with CH(3)OH concentration, and the enhancement coefficient (k'), defined by k(obs) = k + k'[CH(3)OH], is found to be (3.90 +/- 1.87) x 10(-35) x exp[(3849 +/- 135)/T] cm(6) molecule(-2) s(-1) at 30 Torr. The analogous water vapor enhancement coefficient (k'') is (1.16 +/- 0.58) x 10(-36) x exp[(4614 +/- 145)/T] cm(6) molecule(-2) s(-1). The pressure-broadened HO(2) absorption cross section is independent of temperature in the range studied. The line center absorption cross sections at 1506.43 nm, after correction for instrumental broadening, are (4.3 +/- 1.1) x 10(-19), (2.8 +/- 0.7) x 10(-19), and (2.0 +/- 0.5) x 10(-19) cm(2)/molecule at total pressures of 0, 30, and 60 Torr, respectively (95% confidence limits). PMID- 20014816 TI - Stripping voltammetric detection of mercury(II) based on a bimetallic Au-Pt inorganic-organic hybrid nanocomposite modified glassy carbon electrode. AB - A new, highly sensitive and selective sensor for the electrochemical assay of Hg(II) by anodic stripping voltammetry has been developed, whereby a glassy carbon electrode is modified with a novel inorganic-organic hybrid nanocomposite, namely, bimetallic Au-Pt nanoparticles/organic nanofibers (labeled as Au PtNPs/NFs). The sensor possesses a three-dimensional (3D) porous network nanoarchitecture, in which the bimetallic Au-Pt NPs serving as metal NP-based microelectrode ensembles are homogenously distributed in the matrix of interlaced organic NFs. The surface structure and composition of the sensor were characterized by scanning electron microscopy (SEM) and X-ray photoelectron spectroscopy (XPS). Its electrochemical performance was systematically investigated. Our results show that such a newly designed, Au-PtNPs/NF nanohybrid modified electrode provides remarkably improved sensitivity and selectivity for the stripping assay of Hg(II). The detection limit is found to be as low as 0.008 ppb (S/N = 3) that is much below the guideline value from the World Health Organization (WHO). Interferences from other heavy metal ions such as Cu(II), Cr(III), Co(II), Fe(II), Zn(II), and Mn(II) ions associated with mercury analysis are effectively inhibited. Toward the goal for practical applications, the sensor was further evaluated by monitoring Hg(II) in tap and river water specimens. PMID- 20014817 TI - Impurity profiling of a chemical weapon precursor for possible forensic signatures by comprehensive two-dimensional gas chromatography/mass spectrometry and chemometrics. AB - In this report we present the feasibility of using analytical and chemometric methodologies to reveal and exploit the chemical impurity profiles from commercial dimethyl methylphosphonate (DMMP) samples to illustrate the type of forensic information that may be obtained from chemical-attack evidence. Using DMMP as a model compound of a toxicant that may be used in a chemical attack, we used comprehensive two-dimensional gas chromatography/time-of-flight mass spectrometry (GC x GC/TOF-MS) to detect and identify trace organic impurities in six samples of commercially acquired DMMP. The GC x GC/TOF-MS data was analyzed to produce impurity profiles for all six DMMP samples using 29 analyte impurities. The use of PARAFAC for the mathematical resolution of overlapped GC x GC peaks ensured clean spectra for the identification of many of the detected analytes by spectral library matching. The use of statistical pairwise comparison revealed that there were trace impurities that were quantitatively similar and different among five of the six DMMP samples. Two of the DMMP samples were revealed to have identical impurity profiles by this approach. The use of nonnegative matrix factorization indicated that there were five distinct DMMP sample types as illustrated by the clustering of the multiple DMMP analyses into five distinct clusters in the scores plots. The two indistinguishable DMMP samples were confirmed by their chemical supplier to be from the same bulk source. Sample information from the other chemical suppliers supported the idea that the other four DMMP samples were likely from different bulk sources. These results demonstrate that the matching of synthesized products from the same source is possible using impurity profiling. In addition, the identified impurities common to all six DMMP samples provide strong evidence that basic route information can be obtained from impurity profiles. Finally, impurities that may be unique to the sole bulk manufacturer of DMMP were found in some of the DMMP samples. PMID- 20014818 TI - Liposomes act as effective biolubricants for friction reduction in human synovial joints. AB - Phospholipids (PL) form the matrix of biological membranes and of the lipoprotein envelope monolayer, and are responsible for many of the unique physicochemical, biochemical, and biological properties of these supermolecular bioassemblies. It was suggested that phospholipids present in the synovial fluid (SF) and on the surface of articular cartilage have major involvement in the low friction of cartilage, which is essential for proper mobility of synovial joints. In pathologies, such as impaired biolubrication (leading to common joint disorders such as osteoarthritis), the level of phospholipids in the SF is reduced. Using a human-sourced cartilage-on-cartilage setup, we studied to what extent and how phospholipids act as highly effective cartilage biolubricants. We found that large multilamellar vesicles (MLV), >800 nm in diameter, composed of 1,2 dimyristoyl-sn-glycero-3-phosphocholine (DMPC) or of a mixture of DMPC and 1,2 dipalmitoyl-sn-glycero-3-phosphocholine (DPPC) are superior lubricants in comparison to MLV composed of other phosphatidylcholines. Introducing cholesterol into liposomes resulted in less effective lubricants. DMPC-MLV was also superior to small unilamellar vesicles (SUV), <100 nm in diameter, composed of DMPC. MLV are superior to SUV due to MLV retention at and near (<200 microm below) the cartilage surface, while SUV penetrate deeper into the cartilage (450-730 microm). Superiority of specific PL compositions is explained by the thermotropic behavior (including compressibility) of the lipid bilayer. Correlating physicochemical properties of the MLV with the friction results suggests that MLV having lipid bilayers in the liquid-disordered phase and having a solid-ordered to liquid-disordered phase transition temperature slightly below physiological temperature are optimal for lubrication. High phospholipid headgroup hydration, high compressibility, and softness are the common denominators of all efficient PL compositions. The high efficiency of DMPC-MLV and DMPC/DPPC-MLV as cartilage lubricants combined with their resistance to degradation at 37 degrees C supports further evaluation of these MLV for treatment of joint impairments related to poor lubrication. This work also demonstrates the relevance of basic physicochemical properties of phospholipids to their activities in biological systems. PMID- 20014819 TI - X-ray diffraction, solution structure, and computational studies on derivatives of (3-sec-butyl-2,3-dihydro-1H-isoquinolin-4-ylidene)acetic acid: compounds with activity as calpain inhibitors. AB - A thorough experimental and computational study of derivatives of (3-sec-butyl 2,3-dihydroisoquinolin-4-ylidene)acetic acid was performed. Some of these compounds are calpain inhibitors and could be useful as therapeutic agents, since this enzyme is a Ca(2+)-dependent cysteine protease involved in a wide variety of metabolic and physiological processes, whose over-activation is associated to several pathological conditions. To gain a better understanding of the structure activity relationships, a structural analysis was carried out with (1)H and (13)C NMR spectroscopy and DFT calculations together with the X-ray diffraction data of three compounds. The solid state structures showed that the crystal packing as well as the intermolecular interactions depend on the substituent nature of the COOR group. Also, the reactivity of the exocyclic double bond was theoretically evaluated, finding that the more reactive compound is the most potent inhibitor of calpain (IC(50) = 25 nM). PMID- 20014820 TI - Carrier profiling of individual Si nanowires by scanning spreading resistance microscopy. AB - Individual silicon nanowires (NWs) doped either by ion implantation or by in situ dopant incorporation during NW growth were investigated by scanning spreading resistance microscopy (SSRM). The carrier profiles across the axial cross sections of the NWs are derived from the measured spreading resistance values and calibrated by the known carrier concentrations of the connected Si substrate or epi-layer. In the case of the phosphorus ion-implanted and subsequently annealed NWs, the SSRM profiles revealed a radial core-shell distribution of the activated dopants. The carrier concentration close to the surface of a phosphorus-doped NW is found to be by a factor of 6-7 higher than the value in the core and on average only 25% of the implanted phosphorus is electrically active. In contrast, for the in situ boron-doped NW the activation rate of the boron atoms is significantly higher than for phosphorus atoms. Some specific features of SSRM experiments of Si NWs are discussed including the possibility of three dimensional measurements. PMID- 20014821 TI - Plasmonic activity of large-area gold nanodot arrays on arbitrary substrates. AB - Highly efficient fabrication of well-ordered, embedded gold nanodot matrices using diffraction mask projection laser ablation is demonstrated. These gold nanodot arrays are ideally generated onto sapphire substrates but do also form onto AlO(x) thin films, enabling the application to arbitrary bulk substrates. Well-ordered gold dots become embedded into the Al(2)O(3) substrate during the process, thus improving their mechanical stability, chemical inertness, and technological compliance. Such substrates may be useful, for example, to enhance solar-cell efficiency by surface plasmons or as convenient, biocompatible focusing elements in nearfield optical tweezers. PMID- 20014823 TI - High-photoluminescence-yield gold nanocubes: for cell imaging and photothermal therapy. AB - Gold nanocubes demonstrate unique optical properties of the high photoluminescence (PL) quantum yield and a remarkably enhanced extinction band at 544 nm. The 4 x 10(-2) PL yield, which is about 200 times higher than that of gold nanorods, allows gold nanocubes to be successfully used in cell imaging of human liver cancer cells (QGY) and human embryo kidney cells (293T) with a common method of single-photon excitation. The high extinction coefficients of gold nanocubes also facilitate them carrying out the photothermal therapy of QGY and 293T cells, showing similar photokilling efficiency as compared to gold nanorods. PMID- 20014824 TI - Size-selective growth and stabilization of small CdSe nanoparticles in aqueous solution. AB - Using cysteine and its derivatives as capping molecules, we investigated the influence of the physical structure and chemical nature of capping molecules on the selective growth and stabilization of small CdSe nanoparticles (NPs) in aqueous solution at room temperature. Our investigations revealed specific roles for each functional group of cysteine, and we could correlate this structure and nature of the capping molecules with the size, size restriction, size distribution, and stability of the NPs. For selective growth and stabilization of the NPs in aqueous solution, their capping molecules should have at least one functional group with strong nucleophilicity as well as another free, charged functional group. Capping molecules acting as a monodentate ligand were more effective than those acting as a bidentate ligand for restricting the NPs to a smaller size, whereas the former was less effective than the latter for getting a narrower NP size distribution. Capping molecules with relatively bulky spatial geometry near the ligand-NP interface resulted in the formation of NPs with poor short- and long-term stabilities, whereas those having relatively compact spatial geometry near the interface led to NPs with at least moderate short-term stability. We saw that capping molecules having relatively compact outermost spatial geometry led to NPs with excellent long-term stability, whereas those having relatively bulky outermost spatial geometry produced NPs with at most only moderate long-term stability. Our results clearly showed general trends for the possibility of selective growth of stable semiconductor NPs with particular sizes in aqueous solution. PMID- 20014822 TI - Synthetic polymer nanoparticles with antibody-like affinity for a hydrophilic peptide. AB - Synthetic polymer nanoparticles with antibody-like affinity for a hydrophilic peptide have been prepared by inverse microemulsion polymerization. Peptide affinity was achieved in part by incorporating the target (imprint) peptide in the polymerization reaction mixture. Incorporation of the imprint peptide assists in the creation of complementary binding sites in the resulting polymer nanoparticle (NP). To orient the imprint peptide at the interface of the water and oil domains during polymerization, the peptide target was coupled with fatty acid chains of varying length. The peptide--NP binding affinities (ca. 90-900 nM) were quantitatively evaluated by a quartz crystal microbalance (QCM). The optimal chain length was established that created high affinity peptide binding sites on the surface of the nanoparticles. This method can be used for the preparation of nanosized synthetic polymers with antibody-like affinity for hydrophilic peptides and proteins ("plastic antibodies"). PMID- 20014825 TI - Arabinogalactan-folic acid-drug conjugate for targeted delivery and target activated release of anticancer drugs to folate receptor-overexpressing cells. AB - Folic acid (FA) is a high affinity ligand (K(d) = 0.1-1 nM) of folate receptors (FRs) responsible for cellular uptake of folates via receptor-mediated endocytosis. FRs are frequently overexpressed in malignant epithelial cells including ovary, brain, kidney, breast, colon, and lung. FR has emerged as a target for the differential-delivery of anticancer chemotherapeutics with several FA-linked therapeutic agents currently undergoing clinical trials. Here we show that by tethering both FA and the anticancer drug methotrexate (MTX) to arabinogalactan (AG), a highly branched natural polysaccharide with unusual water solubility, a targeted biomacromolecular nanovehicle is formed, which can differentially deliver a cytotoxic cargo into FR-overexpressing cells. Moreover, by linking MTX via an endosomally cleavable peptide (GFLG), we demonstrate a target-activated release mechanism. This FA-AG-GFLG-MTX drug conjugate displayed 6.3-fold increased cytotoxic activity to FR-overexpressing cells compared to their FR-lacking counterparts. These findings establish a novel FA-tethered polymeric nanoconjugate for the targeted delivery of antitumor agents into cancer cells overexpressing FR. PMID- 20014826 TI - Total solid-phase synthesis of NOTA-functionalized peptides for PET imaging. AB - A convenient approach to functionalize peptides either at the N-terminal or on a lysine side chain with 1,4,7-triazacyclononane-N,N',N''-triacetic acid (NOTA) chelating unit has been developed on solid support. The chelate was assembled in a two-step process starting with bromo-acetylated peptides. Deprotection and cleavage of the resin-bound NOTA peptides were performed with use of trifluoroacetic acid (TFA) in the presence of thioanisole and water to give free NOTA peptides. PMID- 20014827 TI - Two-photon absorption property of a conjugated polymer: influence of solvent and concentration on its property. AB - The Suzuki coupling reaction was utilized to prepare target conjugated polymer P1, whose main chain consists of 3,5-dicyanopyridine moieties as electron acceptors and both triphenylamino groups and fluorene moieties as donors. This polymer exhibits excellent solubility in organic solvents and high thermal stability (5% of weight loss at 398 degrees C). Detailed spectroscopic studies including absorption and fluorescence emission in solvents of different polarity were also conducted. The influence of the solvent and concentration on its two photon absorption (TPA) property was studied by a two-photon-induced fluorescence (TPIF) method. It was found that the highest two-photon absorption cross section (13,260 GM at a laser wavelength of 940 nm) appeared in 1,2-dichloroethane, a solvent with a moderate polarity. Meanwhile, with the increase of its concentration in chloroform, its TPA cross section value is reduced, implying that its two-photon absorption process in its chloroform solution is not a pure intrachain process. PMID- 20014828 TI - Sorption mechanisms of arsenate during coprecipitation with ferrihydrite in aqueous solution. AB - Dilute arsenate (As(V)) coprecipitation by ferrihydrite was investigated to determine if treatment of acid mine drainage containing dilute As(V) using coprecipitation is feasible. The sorption density obtained at pH 5 and 7 was nearly identical when As(V) was coprecipitated with ferrihydrite, while it was higher at pH 5 when As(V) was adsorbed on the ferrihydrite. The high sorption density of As(V) to ferrihydrite in coprecipitation with 1-h reaction time suggested that coprecipitation occurs via both adsorption and precipitation. Furthermore, the relationship between residual As(V) and sorption density revealed a BET-type isotherm, with a transition point from a low residual As(V) concentration to a high residual As(V) concentration being observed for all initial As(V) concentrations between 0.15 and 0.44 mmol/dm(3) when the initial molar ratio was 0.56 at pH 5 and 7. X-ray diffraction and the zeta potential revealed that the transition point from surface complexation to precipitation was obtained when the initial As/Fe ratio was 0.4 or 0.5. When dilute As(V) was coprecipitated with ferrihydrite at pH 5 and 7, it was primarily adsorbed as a surface complex when the initial molar ratio was As/Fe < 0.4, while a ferric arsenate and surface complex was formed when this ratio was >or= 0.4. PMID- 20014829 TI - Potentiation of the human GABA(A) receptor as a novel mode of action of lower chlorinated non-dioxin-like PCBs. AB - PCBs are still ubiquitous pollutants despite the ban on their industrial and commercial use. To date, risk characterization and assessment of non-dioxin-like PCBs (NDL-PCBs), especially with respect to neurotoxicity, is hampered by a lack of data. Therefore, the effects of six common NDL congeners (PCB28, 52, 101, 138, 153 and 180) on human GABA(A) receptors, expressed in Xenopus oocytes, were investigated using the two-electrode voltage-clamp technique. When coapplied with GABA (at EC(20)), PCB28 and PCB52 concentration-dependently potentiate the GABA(A) receptor-mediated ion current. Though the LOEC for both PCB28 and PCB52 is 0.3 microM, PCB28 is more potent than PCB52 (maximum potentiation at 10 muM amounting to 98.3 +/- 12.5% and 25.5 +/- 1.4%, respectively). Importantly, coapplication of PCB28 (0.3 microM) and PCB52 (10 microM) resulted in an apparently additive potentiation of the GABA(A) response, whereas coapplication of PCB28 (0.3 microM) and PCB153 (10 microM) attenuated the PCB28-induced potentiation. The present results suggest that the potentiation of human GABA(A) receptor function is specific for lower-chlorinated NDL-PCBs and that higher molecular weight PCBs may attenuate this potentiation as a result of competitive binding to human GABA(A) receptors. Nonetheless, this novel mode of action could (partly) underlie the previously recognized NDL-PCB-induced neurobehavioral alterations. PMID- 20014830 TI - Synthesis and reaction of [(Tp(iPr2))LnH2]3 (Ln = Y, Lu) with CO: trinuclear cluster-bound propenolate en route to selective formation of propene. AB - The use of the Tp(iPr(2)) ligand led to the straightforward and high-yield synthesis of rare examples of trinuclear monoligand lanthanide dihydrides, [(Tp(iPr(2)))LnH(2)](3) (Ln = Y, Lu). The Y complex was found to mediate the hydrogenation and coupling of carbon monoxide with exclusive formation of propene via the intermediacy of a cluster-bound propenolate ligand. PMID- 20014831 TI - Binding of coenzyme B induces a major conformational change in the active site of methyl-coenzyme M reductase. AB - Methyl-coenzyme M reductase (MCR) is the key enzyme in methane formation by methanogenic Archaea. It converts the thioether methyl-coenzyme M and the thiol coenzyme B into methane and the heterodisulfide of coenzyme M and coenzyme B. The catalytic mechanism of MCR and the role of its prosthetic group, the nickel hydrocorphin coenzyme F(430), is still disputed, and no intermediates have been observed so far by fast spectroscopic techniques when the enzyme was incubated with the natural substrates. In the presence of the competitive inhibitor coenzyme M instead of methyl-coenzyme M, addition of coenzyme B to the active Ni(I) state MCR(red1) induces two new species called MCR(red2a) and MCR(red2r) which have been characterized by pulse EPR spectroscopy. Here we show that the two MCR(red2) signals can also be induced by the S-methyl- and the S trifluoromethyl analogs of coenzyme B. (19)F-ENDOR data for MCR(red2a) and MCR(red2r) induced by S-CF(3)-coenzyme B show that, upon binding of the coenzyme B analog, the end of the 7-thioheptanoyl chain of coenzyme B moves closer to the nickel center of F(430) by more than 2 A as compared to its position in both, the Ni(I) MCR(red1) form and the X-ray structure of the inactive Ni(II) MCR(ox1 silent) form. The finding that the protein is able to undergo a conformational change upon binding of the second substrate helps to explain the dramatic change in the coordination environment induced in the transition from MCR(red1) to MCR(red2) forms and opens the possibility that nickel coordination geometries other than square planar, tetragonal pyramidal, or elongated octahedral might occur in intermediates of the catalytic cycle. PMID- 20014833 TI - Zero thermal expansion in a flexible, stable framework: tetramethylammonium copper(I) zinc(II) cyanide. AB - Tetramethylammonium copper(I) zinc(II) cyanide, which consists of N(CH(3))(4)(+) ions trapped within a cristobalite-like metal cyanide framework, has been studied by variable-temperature powder and single-crystal X-ray diffraction. Its coefficient of thermal expansion is approximately zero over the temperature range 200-400 K and comparable with the best commercial zero thermal expansion materials. The atomic displacement parameters, apparent bond lengths, and structure of a low-temperature, low-symmetry phase reveal that the low-energy vibrational modes responsible for this behavior maintain approximately rigid Zn coordination tetrahedra but involve significant distortion of their Cu counterparts. PMID- 20014832 TI - The length of the bound fatty acid influences the dynamics of the acyl carrier protein and the stability of the thioester bond. AB - Acyl carrier proteins involved in fatty acid biosynthesis have been shown to exhibit a high degree of conformational flexibility, in that they are able to sequester fatty acid intermediates between 4 and 18 carbons in length. This flexibility has been observed in X-ray and NMR structures of acyl carrier proteins attached to different fatty acids. NMR studies comparing decanoyl-ACP and stearoyl-ACP indicated that ACP exhibits more dynamic motions when bound to longer fatty acids. We have used complementary chemical and NMR methods as an approach to improving our understanding of the effect of fatty acid length on the dynamics of acyl carrier protein. A chemical assay of the accessibility of the acyl thioester to solvent revealed a positive correlation between chain length and rate of hydrolysis. Surprisingly, this linear correlation was biphasic, with accelerated hydrolysis observed for fatty acids longer than 15 carbons. To further understand the motions associated with this acceleration, we collected (15)N relaxation dispersion data for 14:0-, 15:0-, and 16:0-ACP. The greatest dispersions were exhibited by residues that form the entrance to the fatty acid binding pocket. In addition, these dispersions were observed to increase with the length of the fatty acid. Because the exchange rates derived from fitting the data to a two-state model varied from residue to residue, a more complex motional model appears to be required to adequately explain the dynamics. Thus, acyl-ACP offers an interesting system for future investigations of complex protein motions on the micro- and millisecond time scales. PMID- 20014834 TI - A 6-mer photocontrolled oligonucleotide as an effective telomerase inhibitor. AB - Telomerase is a relevant target for cancer therapy. A small 6-mer psoralen containing oligonucleotide is shown to cause efficient photo-cross-linking to human telomere DNA by G-quadruplex formation. The small, photocontrolled oligonucleotide shows effective telomerase inhibition, both in vitro and in vivo, only when light irradiated. Data suggest that it is a promising agent for the treatment of cancer. PMID- 20014835 TI - Long-range charge transfer through DNA by replacing adenine with diaminopurine. AB - A positive charge migrates along DNA mainly via a series of short-range charge transfer (CT) processes between G-C base pairs, which have relatively high HOMO levels. As such, the CT efficiency sharply decreases with the insertion of A-T base pairs between the G-C base pairs. We have previously demonstrated that the CT efficiency through DNA can be dramatically increased by using deazaadenine (Z), an analogue of A, to adjust the HOMO levels of the A-T base pairs closer to those of the G-C base pairs (Nat. Chem. 2009, 1, 156). In the present study, we have expanded this approach to show that the CT efficiency can also be increased by replacing A bases with diaminopurine (D). PMID- 20014837 TI - Probing the limits of the majority-rules principle in a dynamic supramolecular polymer. AB - By systematic variation of the chemical structure of benzene-1,3,5-tricarboxamide (BTA) derivatives, the effect of chemical structure on the amplification of chirality was studied and quantified. In combination with temperature-dependent amplification experiments, the limits of the majority-rules principle were also investigated. For all BTA derivatives a high, constant helix reversal penalty was determined, which is related to the intermolecular hydrogen bonds that are present in all studied derivatives. For asymmetrically substituted BTA derivatives an odd-even effect was found in the degree of chiral amplification when changing the position of the stereogenic center with respect to the amide functionality. It was found that the mismatch penalty could be directly related to the number of stereocenters present in the molecules. Increasing this number from one to three resulted in an increase in this energy penalty while leaving the helix reversal penalty unaffected. For the majority-rules principle this implies that a single stereocenter present in the molecule contains sufficient chiral information at the molecular level to result in a chirally amplified state at the supramolecular level. Further evidence that the mismatch penalty is directly related to the number of stereocenters was obtained from mixed majority rules experiments where two BTA derivatives with different numbers of stereocenters with opposite stereoconfiguration were studied in a majority-rules experiment. Finally, the ultimate limits of chiral amplification for the majority rules principle were investigated, revealing that, given a certain helix reversal penalty, there is an optimum to which the mismatch penalty can be reduced while also enhancing the degree of chiral amplification. Temperature-dependent majority rules experiments could indeed confirm these simulations. These findings show the relevance of both energy penalties when trying to enhance the degree of chiral amplification for the majority-rules principle in a one-dimensional helical supramolecular polymer. PMID- 20014838 TI - Tuning the extent of chiral amplification by temperature in a dynamic supramolecular polymer. AB - Here, we report on the strong amplification of chirality observed in supramolecular polymers consisting of benzene-1,3,5-tricarboxamide monomers and study the chiral amplification phenomena as a function of temperature. To quantify the two chiral amplification phenomena, i.e., the sergeants-and-soldiers principle and the majority-rules principle, we adapted the previously reported sergeants-and-soldiers model, which allowed us to describe both amplification phenomena in terms of two energy penalties: the helix reversal penalty and the mismatch penalty. The former was ascribed to the formation of intermolecular hydrogen bonds and was the larger of the two. The latter was related to steric interactions in the alkyl side chains due to the stereogenic center. With increasing temperature, the helix reversal penalty was little affected and remained rather constant, showing that the intermolecular hydrogen bonds remain intact and are directing the helicity in the stack. The mismatch penalty, however, was found to decrease when the temperature was increased, which resulted in opposite effects on the degree of chiral amplification when comparing the sergeants-and-soldiers and the majority-rules phenomena. While for the former a reduction in mismatch penalty resulted in a decrease in degree of chiral amplification, for the latter it resulted in a stronger chiral amplification effect. By combining the sergeants-and-soldiers and majority-rules phenomena in a diluted majority-rules experiment, we could further determine the effect of temperature on the degree of chiral amplification. Extending the experiments to different concentrations revealed that the relative temperature, i.e., the temperature relative to the critical temperature of elongation, controls the degree of chiral amplification. On the basis of these results, it was possible to generate a general "master curve" independent of concentration to describe the temperature-dependent majority-rules principle. As a result, unprecedented expressions of amplification of chirality are recorded. PMID- 20014839 TI - Highly permeable zeolite imidazolate framework-8 membranes for CO2/CH4 separation. AB - ZIF-8 membranes, a type of zeolite imidazolate framework, were synthesized by secondary seeded growth on tubular alpha-Al(2)O(3) porous supports. The presence of small, highly crystalline, microporous crystals with narrow particle size distribution led to continuous thin membranes. The synthesized novel ZIF-8 membranes displayed unprecedented high CO(2) permeances and relatively high separation indexes for equimolar mixtures of CO(2) and CH(4). PMID- 20014840 TI - Astatine standard redox potentials and speciation in acidic medium. AB - A combined experimental and theoretical approach is used to define astatine (At) speciation in acidic aqueous solution and to answer the two main questions raised from literature data: does At(0) exist in aqueous solution and what is the chemical form of At(+III), if it exists. The experimental approach considers that a given species is characterized by its distribution coefficient (D) experimentally determined in a biphasic system. The change in speciation arising from a change in experimental conditions is observed by a change in D value. The theoretical approach involves quasi-relativistic quantum chemistry calculations. The results show that At at the oxidation state 0 cannot exist in aqueous solution. The three oxidation states present in the range of water stability are At(-I), At(+I), and At(+III) and exist as At(-), At(+), and AtO(+), respectively, in the 1-2 pH range. The standard redox potentials of the At(+)/At(-) and AtO(+)/At(+) couples have been determined, the respective values being 0.36 +/- 0.01 and 0.74 +/- 0.01 V vs NHE. PMID- 20014841 TI - Inverse CeO2/CuO catalyst as an alternative to classical direct configurations for preferential oxidation of CO in hydrogen-rich stream. AB - A novel inverse CeO(2)/CuO catalyst for preferential oxidation of CO in H(2)-rich stream (CO-PROX) has been developed on the basis of a hypothesis extracted from previous work of the group (JACS 2007, 129, 12064). Possible separation of the two competing oxidation reactions involved in the process (of CO and H(2), respectively) is the key to modulation of overall CO-PROX activity and is based on involvement of different sites as most active ones for each of the two reactions. Achievement of large size CuO particles and adequate CeO(2)-CuO interfacial configurations in the inverse catalyst apparently allows appreciable enhancement of the catalytic properties of this kind of system for CO-PROX, constituting an interesting alternative to classic direct configurations so far explored for this process. Reasons for such behavior are analyzed on the basis of operando-XRD, -XAFS, and -DRIFTS studies. PMID- 20014842 TI - Ligandless, anionic, arylpalladium halide intermediates in the Heck reaction. AB - We report the isolation and reactivity of a series of "ligandless," anionic arylpalladium complexes of the general structure [Pd(Ar)Br(2)](2)(2-) by the reaction of ((t)Bu(3)P)Pd(Ar)(Br) and bromide. These anionic complexes insert olefins at room temperature, and these fast insertions indicate that the anionic complexes are kinetically competent to be intermediates in Heck-Mizoroki reactions conducted under "ligandless" conditions (lacking added dative ligand). Kinetic studies showed that the anionic complexes insert olefins much faster than the corresponding neutral, P(t-Bu)(3)-ligated complexes. Addition of halide to the reaction of the neutral complex ((t)Bu(3)P)Pd(Ar)(Br) and styrene led to a significant rate acceleration, suggesting that the anionic complex forms rapidly and reversibly in situ from the neutral species prior to migratory insertion. These data, along with studies on the regioselectivity for reaction of aryl halides with butyl vinyl ether in the presence of the different starting catalysts, are consistent with the intermediacy of the same anionic, arylpalladium intermediates in Heck reactions catalyzed by palladium complexes containing bulky trialkylphosphine ligands as in reactions conducted under ligandless conditions. PMID- 20014843 TI - Larger spontaneous polarization ferroelectric inorganic-organic hybrids: [PbI3](infinity) chains directed organic cations aggregation to Kagome-shaped tubular architecture. AB - Four isostructural inorganic-organic hybrid ferroelectric compounds, assembled from achiral 3-R-benzylidene-1-aminopyridiniums (R = NO(2), Br, Cl, or F for 1-4, respectively) and [PbI(3)](-) anions with the chiral Kagome-shaped tubular aggregating architecture, show larger spontaneous polarizations. PMID- 20014844 TI - Direct probing of spin state dynamics coupled with electronic and structural modifications by picosecond time-resolved XAFS. AB - The first direct observation of the transient spin-state in a disordered magnetic system with time-resolved XAFS is reported. By observing the evolution of the Fe(II) 1s-3d transition, the spin crossover transition from the (1)A(1) low spin state to (5)T(2) high spin state has been directly observed on a picosecond time scale. Moreover, observation of the transient spin state with time-resolved XAFS allows for the investigation of the variations in the electronic state and molecular structure. This unique experimental technique probes the excited states involved in the ultrafast photoinduced reactions in disordered magnetic systems. PMID- 20014845 TI - Optical imaging of mammary and prostate tumors in living animals using a synthetic near infrared zinc(II)-dipicolylamine probe for anionic cell surfaces. AB - In vivo optical imaging shows that a fluorescent imaging probe, comprised of a near-infrared fluorophore attached to an affinity group containing two zinc(II) dipicolylamine (Zn-DPA) units, targets prostate and mammary tumors in two different xenograft animal models. The tumor selectivity is absent with control fluorophores whose structures do not have appended Zn-DPA targeting ligands. Ex vivo biodistribution and histological analyses indicate that the probe is targeting the necrotic regions of the tumors, which is consistent with in vitro microscopy showing selective targeting of the anionic membrane surfaces of dead and dying cells. PMID- 20014847 TI - Role of pyridoxamine in the formation of the Amadori/Heyns compounds and aggregates during the glycation of beta-lactoglobulin with galactose and tagatose. AB - The effect of pyridoxamine on the Maillard reaction during the formation of conjugates of beta-lactoglobulin with galactose and tagatose under controlled conditions (pH 7, 0.44 aw, 40 and 50 degrees C, for 6 days) has been studied, for the first time, by means of the changes in reducing carbohydrates, formation of Amadori or Heyns compounds, and aggregates and browning development. The results showed the formation of interaction products between pyridoxamine and galactose or tagatose either in the presence or in the absence of beta-lactoglobulin, indicating that pyridoxamine competes with the free amino groups of beta lactoglobulin for the carbonyl group of both carbohydrates. Thus, a small inhibitory effect of pyridoxamine on the initial stages of the Maillard reaction was pointed out. Furthermore, much lower aggregation and color formation rates were observed in the conjugates of beta-lactoglobulin galactose/tagatose with pyridoxamine than without this compound, supporting its potent inhibitory effect on the advanced and final stages of the Maillard reaction. These findings reveal the usefulness of food-grade inhibitors of the advanced stages of the Maillard reaction, such as pyridoxamine, that, in combination with mild storage conditions, could lead to the formation of safer neoglycoconjugates without impairing their nutritional quality. PMID- 20014846 TI - Differential tissue distribution of sesaminol triglucoside and its metabolites in rats fed with lignan glycosides from sesame meal with or without nano/submicrosizing. AB - Lignan glycosides are important functional compounds in sesame meal. In the present study, we investigated whether the tissue distribution of nano/submicrosized lignan glycosides from sesame meal (N-LGSM) differs from lignan glycosides from sesame meal (LGSM). LGSM was nano/submicrosized with 0.3 mm zirconia beads as the milling media. The average particle size of the 4% LGSM aqueous suspension reduced rapidly from approximately 2 microm to 200 nm after media milling at an agitation speed of 3600 rpm for 30 min. We examined the tissue distribution of sesaminol triglucoside (ST), the main component in LGSM, in Sprague-Dawley (SD) rats. The concentrations of ST were determined in various tissues and plasma within a 24 h period after oral administration of N-LGSM and LGSM (800 mg/kg of body weight). The results showed that higher concentrations of ST and its metabolites (sesaminol, sesaminol sulfate, and sesaminol glucuronide) were found in N-LGSM compared to those in LGSM in most tissues, especially liver and small intestine. Sesaminol glucuronide was the main metabolite in rats. After 3 h of oral administration, around 70% higher concentration of sesaminol glucuronide was found in N-LGSM compared to that in LGSM. This study clearly showed that LGSM is more bioavailable after nano/submicrosizing. PMID- 20014848 TI - Thermal stability of thaumatin-like protein, chitinase, and invertase isolated from Sauvignon blanc and Semillon juice and their role in haze formation in wine. AB - A thermal unfolding study of thaumatin-like protein, chitinase, and invertase isolated from Vitis vinifera Sauvignon blanc and Semillon juice was undertaken. Differential scanning calorimetry demonstrated that chitinase was a major player in heat-induced haze in unfined wines as it had a low melt temperature, and aggregation was observed. The kinetics of chitinase F1 (Sauvignon blanc) unfolding was studied using circular dichroism spectrometry. Chitinase unfolding conforms to Arrhenius behavior having an activation energy of 320 kJ/mol. This enabled a predictive model for protein stability to be generated, predicting a half-life of 9 years at 15 degrees C, 4.7 days at 30 degrees C, and 17 min at 45 degrees C. Circular dichroism studies indicate that chitinase unfolding follows three steps: an initial irreversible step from the native to an unfolded conformation, a reversible step between a collapsed and an unfolded non-native conformation, followed by irreversible aggregation associated with visible haze formation. PMID- 20014849 TI - Assessment of the bioaccessibility of polybrominated diphenyl ethers in foods and the correlations of the bioaccessibility with nutrient contents. AB - Bioaccessibility of a contaminant in foods is the fraction of the contaminant mobilized from the food matrices during gastrointestinal digestion. In the present paper, the bioaccessibility of polybrominated diphenyl ethers (PBDEs) in 13 types of foods, including fish, meat, rice, flour, and vegetables, was determined using an in vitro digestion method. The bioaccessibility obtained ranged from 2.6% to 41.3%. It was found that the bioaccessibility of PBDEs exhibited positive correlations with fat and carbohydrate contents and negative correlations with protein and dietary fiber contents in the foods. Fat was the most important factor affecting the bioaccessibility of PBDEs. The mechanism underlying the correlations was discussed in view of partition of PBDEs between liquid and solid phases. To our knowledge, this is the first article systematically investigating the bioaccessibility of PBDEs in foods and demonstrating the influence of the food constituents on the bioaccessibility of PBDEs. PMID- 20014850 TI - Activation of molecular oxygen by a dioxygenase pathway by a ruthenium bis bipyridine compound with a proximal selenium site. AB - A ruthenium(II) bipyridine complex with proximal phenylselenium tethers, [Ru](H(2)O)(2), reacted intramolecularly with O(2) in a protic slightly acidic solvent, 1,1,1,3,3,3-hexafluoro-2-propanol (HFIP), to yield an O-O bond cleaved product, [Ru](O)(2), with formation of two Ru-O-Se moieties. This stable compound was isolated, and its structure was determined by X-ray diffraction. The identification of the compound in solution was confirmed by ESI-MS and the (1)H NMR with the associated Curie plot that showed that [Ru](O)(2) was paramagnetic. The magnetic susceptibility was 2.8 mu(B) by Evan's method suggesting a ground state triplet or biradical. DFT calculations, however, predicted a ground state singlet and an oxidized Se atom. Further it was shown that [Ru](O)(2) is a potent oxygen transfer species of both O(2)-derived atoms to triphenylphosphine and a nucleophilic alkene such as 2,3-dimethyl-2-butene in both HFIP and acetonitrile. UV-vis spectroscopy combined with the measured stoichiometry of PPh(3):O(2) = approximately 2 in a catalytic oxidation of PPh(3) suggests a dioxygenase type activation of O(2) with structural identification of the O-O bond cleavage reaction step, formation of [Ru](O)(2) as an intermediate, and the proof that [Ru](O)(2) is a donor of both oxygen atoms. PMID- 20014851 TI - Production of fructooligosaccharides by beta-fructofuranosidases from Aspergillus oryzae KB. AB - Aspergillus oryzae KB produces two types of beta-fructofuranosidases: F1 and F2. F1 produces the fructooligosaccharides (FOSs) 1-kestose, nystose, and fructosyl nystose from sucrose through a transfructosylation action, whereas F2 mainly hydrolyzes sucrose to glucose and fructose. F1 and F2 enzymes were more selectively produced from the KB strain in liquid media with a sucrose concentration>2% and <2%, respectively. Immobilization using an anion-exchange resin (WA-30; polystyrene with tertiary amine) and cross-linking with glutaraldehyde depressed the hydrolysis reaction of F2 (high hydrolyzing enzyme) alone and enhanced the thermal stability of F1 (high transferring enzyme). F1 enzyme produced in the high sucrose medium was immobilized, cross-linked, and packed in a tubular reactor for continuous production of FOSs (24.6% 1-kestose, 21.6% nystose, 5.7% and fructosyl nystose). In a long-term operation in which 60% sucrose was imputed at 55 degrees C, the composition of FOSs produced was 51.9% (transfer ratio: 92%), and production by the immobilized enzyme was maintained for 984 h. PMID- 20014852 TI - Analysis of a Nicotiana tabacum L. genomic region controlling two leaf surface chemistry traits. AB - cis-Abienol and sucrose esters are Nicotiana tabacum leaf surface components that likely influence plant resistance to pests. Their breakdown products also contribute to flavor and aroma characteristics of certain tobacco types. Mapping of genes involved in the biosynthesis of these compounds could permit development of molecular-based tools for generating tobacco types with novel cured leaf chemistry profiles. A doubled haploid mapping population segregating for major genes (Abl and BMVSE) affecting the ability to accumulate cis-abienol and sucrose esters was generated and genotyped with a large set of microsatellite markers. The two genes were found to reside on chromosome A of the N. tabacum genome with a distance of 8.2 cM (centimorgans) between them. Seventeen microsatellite markers were also placed on this linkage group, several of which exhibited complete cosegregation with Abl and BMVSE. Results should aid breeding efforts focused on modification of this aspect of tobacco cured leaf chemistry. PMID- 20014853 TI - Oligothiophene template effects on packings and orientations of C60 molecules on Ag(111) surface. AB - The packing conformations of sexithiophene (6T) and the orientations of the C(60) molecules on top of the preadsorbed 6T monolayer on Ag(111) surface have been investigated by the molecular dynamics simulations (on the basis of molecular mechanics) in conjunction with quantum mechanics calculations of the relative strength of intermolecular and interfacial interactions. It is demonstrated that the flat-lying oligothiophene (nT, n = 4 and 6) monolayer is formed on the Ag(111) surface, and the arrangement of 6T molecules is more ordered than that in 4T film. It is also shown that the underlying 6T stripes make C(60) molecules aggregate in chainlike arrays on the 6T covered Ag(111) surface, showing significant template effects on the directed self-assembly of C(60) cages. For the absorbed C(60) molecule on the 6T prepatterned Ag(111) surface, four typical orientations, hexagon, pentagon, 6:6 bond, and 5:6 bond, are found to appear with populations of 26.3%, 2.7%, 37.5%, and 18.8%, respectively. When the deposition order is changed, the 6T stripes are shown to tilt with corrugation on the underlying C(60) carpet, revealing the important role of the deposition order in modulation of the ordered supramolecular nanostructures. PMID- 20014854 TI - Spatial and developmental profiling of miraculin accumulation in transgenic tomato fruits expressing the miraculin gene constitutively. AB - We previously developed a transgenic tomato that expresses the miraculin gene using a constitutive promoter. In this study, we profiled the developmental and spatial accumulation of the miraculin protein and mRNA in transgenic tomato fruits. Miraculin mRNA expression was almost constant up to orange stage, and then the expression increased at red stage. The miraculin protein accumulated gradually during fruit development and reached its highest level at the overripe stage. At the red stage of fruit, miraculin protein was accumulated at the highest level in the exocarp, and similar in other fruit tissues: mesocarp, dissepiment, upper placenta, lower placenta and jelly. Moreover, the pattern of miraculin accumulation in fruit tissues was the same regardless of genetic background and position at which the miraculin gene was inserted in the genome. We also discuss suitable tomato types expressing miraculin for their commercial use. PMID- 20014855 TI - Quantitative comparison of free and bound volatiles of two commercial tomato cultivars (Solanum lycopersicum L.) during ripening. AB - The changes in the profile of both free and glycosidically bound volatiles were studied in Moneymaker and Raf tomato cultivars during fruit ripening. The concentrations of 20 of 24 and of 27 of 30 compounds detected in the free volatile fraction (FVF) and glycosidically bound fraction (GBF), respectively, differed significantly between cultivars during ripening. Most free and bound volatiles increased during ripening in both cultivars. The contribution of each free volatile compound to the overall aroma was estimated by calculating its log U value, which indicated that only 11 compounds seem to exert a strong influence. Nine volatiles were detected exclusively in the GBF, among them geraniol, beta citronellol, alpha-terpineol, and trans- and cis-linalool oxides. Nine other compounds were found to be more abundant in the GBF than in the FVF, their absolute levels varying between cultivars and stages of ripening. According to the log U values reached, of these nine compounds, linalool, 3-methyl-1-butanol, trans-2-hexenal, eugenol, and 2-phenylethanol may have an impact on tomato aroma upon release from their glycosidic conjugates. Sugars resulting from the enzymatic hydrolysis of the GBF were detected by high-performance liquid chromatography. Rhamnose was the most abundant followed by arabinose, glucose, and xylose. PMID- 20014856 TI - Oral bioavailability, urinary excretion and organ distribution of melamine in Sprague-Dawley rats by high-performance liquid chromatography with tandem mass spectrometry. AB - High-performance liquid chromatography with tandem mass spectrometry (HPLC/MS/MS) was used to determine melamine oral bioavailability (BA) and urinary excretion. Organ distribution after a 14-day consecutive oral melamine administration (100 mg/kg/day, once a day) was also evaluated. A noncompartmental model was utilized to obtain pharmacokinetic parameters. According to the results, the BA of melamine was estimated to be 98.1%. Approximately 63% of administered melamine was recovered in urine within 96 h after a single oral administration (100 mg/kg). The bladder had the highest melamine concentration of all the organs after a 14-day consecutive oral administration of melamine, and almost no melamine was found in the rat brain. This result indicated that the oral absorption of melamine was almost complete and urinary excretion was the major route for its elimination. Repeated exposure to high-dose melamine may result in only slight accumulation in organs. PMID- 20014857 TI - Synthesis and antiplasmodial activity of new indolone N-oxide derivatives. AB - A series of 66 new indolone-N-oxide derivatives was synthesized with three different methods. Compounds were evaluated for in vitro activity against CQ sensitive (3D7), CQ-resistant (FcB1), and CQ and pyrimethamine cross-resistant (K1) strains of Plasmodium falciparum (P.f.), as well as for cytotoxic concentration (CC(50)) on MCF7 and KB human tumor cell lines. Compound 26 (5 methoxy-indolone-N-oxide analogue) had the most potent antiplasmodial activity in vitro (<3 nM on FcB1 and = 1.7 nM on 3D7) with a very satisfactory selectivity index (CC(50) MCF7/IC(50) FcB1: 14623; CC(50) KB/IC(50) 3D7: 198823). In in vivo experiments, compound 1 (dioxymethylene derivatives of the indolone-N-oxide) showed the best antiplasmodial activity against Plasmodium berghei, 62% inhibition of the parasitaemia at 30 mg/kg/day. PMID- 20014858 TI - Multivalent benzoboroxole functionalized polymers as gp120 glycan targeted microbicide entry inhibitors. AB - Microbicides are women-controlled prophylactics for sexually transmitted infections. The most important class of microbicides target HIV-1 and contain antiviral agents formulated for topical vaginal delivery. Identification of new viral entry inhibitors that target the HIV-1 envelope is important because they can inactivate HIV-1 in the vaginal lumen before virions can come in contact with CD4+ cells in the vaginal mucosa. Carbohydrate binding agents (CBAs) demonstrate the ability to act as entry inhibitors due to their ability to bind to glycans and prevent gp120 binding to CD4+ cells. However, as proteins they present significant challenges in regard to economical production and formulation for resource-poor environments. We have synthesized water-soluble polymer CBAs that contain multiple benzoboroxole moieties. A benzoboroxole-functionalized monomer was synthesized and incorporated into linear oligomers with 2 hydroxypropylmethacrylamide (HPMAm) at different feed ratios using free radical polymerization. The benzoboroxole small molecule analogue demonstrated weak affinity for HIV-1BaL gp120 by SPR; however, the 25 mol % functionalized benzoboroxole oligomer demonstrated a 10-fold decrease in the K(D) for gp120, suggesting an increased avidity for the multivalent polymer construct. High molecular weight polymers functionalized with 25, 50, and 75 mol % benzoboroxole were synthesized and tested for their ability to neutralize HIV-1 entry for two HIV-1 clades and both R5 and X4 coreceptor tropism. All three polymers demonstrated activity against all viral strains tested with EC(50)s that decrease from 15000 nM (1500 microg mL(-1)) for the 25 mol % functionalized polymers to 11 nM (1 microg mL(-1)) for the 75 mol % benzoboroxole-functionalized polymers. These polymers exhibited minimal cytotoxicity after 24 h exposure to a human vaginal cell line. PMID- 20014859 TI - Color-tuning mechanism of firefly investigated by multi-configurational perturbation method. AB - This is the first report on a multiconfigurational reference second-order perturbation theory-molecular mechanics study of the color modulation of the observed bioluminescence of the oxyluciferin-luciferase complex of the Japanese genji-botaru firefly using structures according to recent X-ray data. Our theoretical results do not support the experimentally deduced conclusion that the color modulation of the emitted light primarily depends on the size of the compact luciferase protein cavity embedding the excited oxyluciferin molecule. Rather, we find, in agreement with recent experimental observations, that the wavelength of the emitted light depends on the polarity of the microenvironment at the phenol/phenolate terminal of the benzothiazole fragment in oxyluciferin. PMID- 20014860 TI - Comparison of extensive protein fractionation and repetitive LC-MS/MS analyses on depth of analysis for complex proteomes. AB - In-depth, reproducible coverage of complex proteomes is challenging because the complexity of tryptic digests subjected to LC-MS/MS analysis frequently exceeds mass spectrometer analytical capacity, which results in undersampling of data. In this study, we used cancer cell lysates to systematically compare the commonly used GeLC-MS/MS (1-D protein + 1-D peptide separation) method using four repetitive injections (2-D/repetitive) with a 3-D method that included solution isoelectric focusing and involved an equal number of LC-MS/MS runs. The 3-D method detected substantially more unique peptides and proteins, including higher numbers of unique peptides from low-abundance proteins, demonstrating that additional fractionation at the protein level is more effective than repetitive analyses at overcoming LC-MS/MS undersampling. Importantly, more than 90% of the 2-D/repetitive protein identifications were found in the 3-D method data in a direct protein level comparison, and the reproducibility between data sets increased to greater than 96% when factors such as database redundancy and use of rigid scoring thresholds were considered. Hence, high reproducibility of complex proteomes, such as human cancer cell lysates, readily can be achieved when using multidimensional separation methods with good depth of analysis. PMID- 20014861 TI - Production of Fumonisin B2 and B4 by Aspergillus niger on grapes and raisins. AB - The recent discovery of fumonisin production in Aspergillus niger, raises concerns about the presence of these mycotoxins in grapes and raisins as well as other commodities where A. niger is a frequent contaminant. Here we investigate the potential production of fumonisins in A. niger cultured on grapes and raisins. Sixty-six A. niger, 4 A. tubingensis, and 16 A. acidus strains isolated from raisins were tested for fumonisin production on laboratory media. Neither A. tubingensis nor A. acidus strains produced fumonisins, but 77% of A. niger strains did. None of the strains produced ochratoxin A. Ten selected fumonisin producing A. niger strains were further able to produce fumonisin B(2) and fumonisin B(4) on grapes in the range 171-7841 microg fumonisin B(2)/kg and 14 1157 microg fumonisin B(4)/kg. Four selected strains were able to produce fumonisin B(2) (5-6476 microg/kg) and fumonisin B(4) (12-672 microg/kg) on raisins. PMID- 20014862 TI - Application of a novel antioxidative assay in beer analysis and brewing process monitoring. AB - A novel antioxidative assay based on direct current polarography has been developed. Quantification of antioxidative (AO) activity has been based on a decrease of hydrogen peroxide anodic current in the presence of antioxidants. An efficient experimental procedure, without any special pretreatment of analyzed samples, has been applied. Antioxidative activity of different kinds of commercial beers (dark, blond, and alcohol-free), some small-scale made special beers with medicinal herbs and mushroom extracts, extracts themselves, as well as individual phenolic components present in beer has been measured. In addition, changes of AO activity during the full-scale industrial process of beer production have been monitored. A strong correlation between results obtained and total phenolics content has been observed. The assay can be recommended for application in brewing industry, either to survey a process with the aim to optimize relevant technological factors or to analyze quality of final product. PMID- 20014863 TI - Homoleptic heavy alkaline Earth and europium triazenides. AB - The sigma-bond metathesis reaction between PhSiH(3) and the heteroleptic metal pentafluorophenyl compounds [Dmp(Tph)N(3)MC(6)F(5)(thf)(n)] (Dmp = 2,6 Mes(2)C(6)H(3) with Mes = 2,4,6-Me(3)C(6)H(2); Tph = 2-TripC(6)H(4) with Trip = 2,4,6-(i)Pr(3)C(6)H(2); n = 1, 2; M = Sr, Ba, Eu) supported by sterically crowded, biphenyl- and terphenyl-substituented triazenido ligands afforded the first homoleptic stontium, barium, and europium triazenides [M{N(3)Dmp(Tph)}(2)] {M = Sr (2), Ba (4), Eu (5)}. Crystallization of 2 from an n-heptane/1,2 dimethoxyethane mixture gave the complex [Sr{N(3)Dmp(Tph)}(2)(dme)] (3). All new compounds have been characterized by (1)H and (13)C NMR spectroscopy (not 5), elemental analysis, IR spectroscopy (5 only), and X-ray crystallography. In the solid-state structures, the first coordination sphere of the metal cations consists of four nitrogen atoms of the two bidentate triazenide ligands. Additional metal-eta(n)-pi-arene-interactions of different hapticity n (n = 3-6) are observed to the flanking arms of the terphenyl substituents. PMID- 20014864 TI - Tailoring of the nanotexture of mesoporous silica films and their functionalized derivatives for selectively harvesting low molecular weight protein. AB - We present a fast, efficient, and reliable system based on mesoporous silica chips to specifically fractionate and enrich the low molecular weight proteome. Mesoporous silica thin films with tunable features at the nanoscale were fabricated using the triblock copolymer template pathway. Using different templates and concentrations in the precursor solution, various pore size distributions, pore structures, and connectivity were obtained and applied for selective recovery of low mass proteins. In combination with mass spectrometry and statistic analysis, we demonstrated the correlation between the nanophase characteristics of the mesoporous silica thin films and the specificity and efficacy of low mass proteome harvesting. In addition, to overcome the limitations of the prefunctionalization method in polymer selection, plasma ashing was used for the first time for the treatment of the mesoporous silica surface prior to chemical modification. Surface charge modifications by different functional groups resulted in a selective capture of the low molecular weight proteins from serum sample. In conclusion, our study demonstrates that the ability to tune the physicochemical properties of mesoporous silica surfaces, for a selective enrichment of the low molecular weight proteome from complex biological fluids, has the potential to promote proteomic biomarker discovery. PMID- 20014866 TI - Evaluation of di-sansalvamide A derivatives: synthesis, structure-activity relationship, and mechanism of action. AB - Described is the SAR of 18 di-sansalvamide A derivatives and the mechanism of action of the most potent compound. We show that this scaffold is a promising lead in the development of novel cancer therapeutics because it is cytotoxic at nanomolar potency, inhibits a well-established oncogenic target (Hsp90), and does not share structural motifs with current drugs on the market. PMID- 20014867 TI - Discovery of a new and efficient small molecule for neuronal differentiation from mesenchymal stem cell. AB - A new synthetic small molecule, compound 1, which induced a neuronal differentiation from mesenchymal stem cells (MSCs) with an excellent efficiency, was identified. Furthermore the differentiated cell by 1 showed the neural electrophysiological and cholinergic neuron properties. PMID- 20014865 TI - Making the cut: the chemical biology of cytokinesis. AB - Cytokinesis is the last step in the cell cycle, where daughter cells finally separate. It is precisely regulated in both time and space to ensure that each daughter cell receives an equal share of DNA and other cellular materials. Chemical biology approaches have been used very successfully to study the mechanism of cytokinesis. In this review, we discuss the use of small molecule probes to perturb cytokinesis, as well as the role naturally occurring small molecule metabolites such as lipids play during cytokinesis. PMID- 20014868 TI - N-sulfonylanthranilic acid derivatives as allosteric inhibitors of dengue viral RNA-dependent RNA polymerase. AB - A novel class of compounds containing N-sulfonylanthranilic acid was found to specifically inhibit dengue viral polymerase. The structural requirements for inhibition and a preliminary structure-activity relationship are described. A UV cross-linking experiment was used to map the allosteric binding site of the compound on the viral polymerase. PMID- 20014869 TI - Janus kinase 2 inhibitors. Synthesis and characterization of a novel polycyclic azaindole. AB - The synthesis and characterization of a novel polycyclic azaindole based derivative is disclosed, and its binding to JAK2 is described. The compound is further evaluated for its ability to block the EPO/JAK2 signaling cascade in vitro and in vivo. PMID- 20014870 TI - Discovery of 6alpha-ethyl-23(S)-methylcholic acid (S-EMCA, INT-777) as a potent and selective agonist for the TGR5 receptor, a novel target for diabesity. AB - In the framework of the design and development of TGR5 agonists, we reported that the introduction of a C(23)(S)-methyl group in the side chain of bile acids such as chenodeoxycholic acid (CDCA) and 6-ethylchenodeoxycholic acid (6-ECDCA, INT 747) affords selectivity for TGR5. Herein we report further lead optimization efforts that have led to the discovery of 6alpha-ethyl-23(S)-methylcholic acid (S EMCA, INT-777) as a novel potent and selective TGR5 agonist with remarkable in vivo activity. PMID- 20014871 TI - Quantum coherence enabled determination of the energy landscape in light harvesting complex II. AB - The near-unity efficiency of energy transfer in photosynthesis makes photosynthetic light-harvesting complexes a promising avenue for developing new renewable energy technologies. Knowledge of the energy landscape of these complexes is essential in understanding their function, but its experimental determination has proven elusive. Here, the observation of quantum coherence using two-dimensional electronic spectroscopy is employed to directly measure the 14 lowest electronic energy levels in light-harvesting complex II (LHCII), the most abundant antenna complex in plants containing approximately 50% of the world's chlorophyll. We observe that the electronically excited states are relatively evenly distributed, highlighting an important design principle of photosynthetic complexes that explains the observed ultrafast intracomplex energy transfer in LHCII. PMID- 20014872 TI - Should financial incentives be used to differentially reward 'me-too' and innovative drugs? AB - Strategies to change the existing mix of innovative and 'me-too' drugs are intended to increase societal value of a given investment in R&D by providing an incentive for firms to invest in drugs that are more likely to be clinically innovative. How can financial incentives be used to change this mix? Will a strategy have its intended consequence or will it have the unintended outcome of reducing the rate at which the population burden of disease is reduced? The perspective of this review is a country such as Australia, Canada or the UK that has universal health insurance and a drug reimbursement process that is informed by economic evidence. A review of the literature was performed and the views of both the proponents and the opponents of such strategies and the mechanisms by which they could be implemented were summarized. The debate is based largely on hypothesized responses by firms to changes in incentives rather than empirical evidence. The main point of contention is whether a changed mix of new molecular entities (NMEs) increases or decreases the total amount of clinical innovation launched each year. The argument presented in this article is that, despite the limited empirical evidence, it is possible to improve our assessment of the likely costs and consequences of a proposed strategy by appealing to economic theory and observations about the reimbursement process. First, the empirical evidence supporting the view that changing a mix of drugs will improve clinical innovation is based on the average launched drug, not the marginal innovative drug otherwise not developed, and therefore could be misleading. Second, the dynamic and complex nature of evidence of clinical innovation will reduce the feasibility of using contractually based mechanisms to implement such a strategy. Also, a single country is unlikely to have an impact on R&D decisions, and variation in the per capita economic value of new drugs would make multi jurisdiction contracts with one firm difficult to implement. Third, the quality of evidence of the clinical innovation of the lead drug could be reduced if there are fewer or no follow-on drugs. Finally, the existing inefficiencies in the process of displacement to finance new technologies from a capped budget reduces the efficiency with which any additional potential clinical innovation from NMEs will be translated to reduced population burden of disease. The article concludes that it is possible that such a strategy could be costly to implement, and the impact on global burden of disease uncertain in both direction and magnitude. This is likely to be the case even if the average clinical innovation content of innovative NMEs is higher than for me-too NMEs and the mechanisms designed to change the mix of NMEs are effective. Other options to improve the effectiveness with which pharmaceutical clinical innovation reduces burden of disease should be explored, including improved efficiency of both firm R&D and the process of disinvestment to finance new technologies. PMID- 20014873 TI - Adverse drug interactions: moving from perception to action. PMID- 20014874 TI - The cost and incidence of prescribing errors among privately insured HIV patients. AB - BACKGROUND: With the rapid growth in the volume of HIV-related studies that address drug interactions, appropriate medication regimens, and when and how to alter drug regimens, it is challenging for physicians to stay informed. Physicians require knowledge about all drugs taken by HIV patients in order to assess accurately the benefits and risks of various drug combinations. OBJECTIVE: To examine the cost and frequency of antiretroviral prescribing errors among a sample of privately insured patients with HIV disease. METHODS: Data were obtained from the MarketScan Commercial Claims and Encounter Database created by the Medstat Group Inc. The MarketScan database contains claims data for inpatient care, outpatient care, physician services and prescription drugs in benefit plans sponsored by >50 large employers in the US. This study compared data from the 1999-2000 MarketScan database with those from the 2005 MarketScan database. The 2005 MarketScan database includes 12,226 HIV enrollees who received antiretroviral drugs. This study compared the claims experience of HIV patients who filled a prescription for a drug combination that is not recommended by the US Department of Health and Human Services Panel on Antiretroviral Guidelines for Adults and Adolescents with the claims experience of patients who did not receive such a prescription. RESULTS: In the 1999-2000 database the most common inappropriate drug combination involved the co-administration of a protease inhibitor (PI) and the lipid-lowering drug simvastatin, and 1% of patients experienced this type of error. In the 2005 database, only 0.4% of patients (46 of 12,226) experienced an inappropriate combination of simvastatin and a PI while 5.3% of patients (644 of 12,226) received atazanavir and tenofovir without ritonavir (referred to herein as 'boosting errors'). Patients who experienced a boosting error incurred higher annual costs than patients who took ritonavir along with tenofovir and atazanavir ($US 20,927 vs $US 16,704). Because atazanavir was approved by the US FDA in June 2003, medication errors involving atazanavir were not relevant in 1999 and 2000. Overall, it was found that HIV patients were three times as likely to experience an inappropriate drug combination in 2005 than they were in either 1999 or 2000 (5.9% vs 1.9%), and that this increase is attributable to boosting errors. In addition, the prevalence rate of HIV in the 2005 MarketScan database was almost triple that in the 1999 MarketScan database (0.14% vs 0.05%). CONCLUSION: This study indicates that those who provide care to HIV patients must be vigilant in their efforts to provide patients with a drug therapy regimen that minimizes the chance of an adverse reaction and maximizes the potential to control viral replication. PMID- 20014875 TI - Cost effectiveness of rituximab maintenance therapy in follicular lymphoma: long term economic evaluation. AB - BACKGROUND: Rituximab maintenance therapy was shown to significantly extend overall survival (OS) and progression-free survival (PFS) in relapsed/refractory follicular lymphoma (FL) in the pivotal EORTC 20981 trial. OBJECTIVE: To assess the long-term costs and cost effectiveness of rituximab maintenance therapy after induction therapy versus current standard practice (observation) from the French National Health Service perspective. METHODS: A lifetime transition model was developed comparing rituximab maintenance with observation. PFS and OS were obtained from the EORTC 20981 trial with a median follow-up of 28 months and extrapolated from 2-year Kaplan-Meier curves using a Weibull distribution. PFS and OS benefits of rituximab were conservatively assumed to last only 5 years. Utility data were obtained from a multicentre observational study using the EQ-5D questionnaire. Direct medical costs were obtained from French official sources. All costs are reported in euro, year 2006 values. RESULTS: The EORTC 20981 study demonstrated that rituximab maintenance was effective in the management of relapsed/refractory FL. The model results showed that life expectancy and QALYs were increased by 22% and 28%, respectively, in patients treated with rituximab. The incremental cost-effectiveness ratios (ICERs) were euro 7612 per life-year gained and euro 8729 per QALY gained. In a one-way sensitivity analysis, most of the ICERs fell within the range of euro 7000-12,000. The results tend to show that rituximab maintenance therapy may be a cost-effective strategy in the management of relapsed/refractory FL in France, with ICERs below those observed for other therapies in the oncology field. The cost of rituximab was partly offset by the lower cost of relapse due to a longer time in the disease-free health state for patients in the rituximab arm. PMID- 20014876 TI - Cost effectiveness of intensive lipid-lowering treatment for patients with congestive heart failure and coronary heart disease in the US. AB - BACKGROUND: A recent study found fewer hospitalizations for congestive heart failure (CHF) patients receiving high-dose versus low-dose statin therapy. OBJECTIVE: To examine the cost effectiveness of high-dose versus low-dose statin therapy in CHF patients. METHODS: Two scenarios (literature-based [base-case scenario] vs trial-based post-event mortality [alternative scenario]) assessed the cost effectiveness of atorvastatin 80 mg/day (A80) versus atorvastatin 10 mg/day (A10) in patients with both CHF and coronary heart disease (CHD) [CHF/CHD], using a lifetime Markov model. The model predicts treatment-specific probabilities of major and minor cardiovascular events and death, based on clinical trial data. The quality of life and costs were literature based. Measures included costs per life-year saved (LYS) and QALY gained. Health consequences and costs were discounted at 3.0% annually. Analyses were conducted from the payer perspective and valued in $US, year 2006-7 values. RESULTS: Literature-based mortality estimates (base case) increased life-years and QALYs for A80 compared with A10 (incremental cost-effectiveness ratios [ICERs]: $US9600 per LYS; $US13 600 per QALY). At a willingness to pay of $US100 000 per QALY, A80 was cost effective in 80% of simulations. A10 dominated A80 when using trial based mortality estimates (alternative scenario). At a willingness to pay of $US100 000 per QALY, A80 was cost effective in 48% of simulations. CONCLUSIONS: Intensive A80 treatment may be cost effective versus A10 in cardiovascular prevention in CHF/CHD patients in the US, due to projected gains in life expectancy and health-related quality of life. However, the results are highly sensitive to assumptions about the mortality rate in the model. When using the mortality rate observed in the trial, A10 dominates A80. PMID- 20014877 TI - A policy model to evaluate the benefits, risks and costs of warfarin pharmacogenomic testing. AB - BACKGROUND: In 2007, the US FDA added information about pharmacogenomics to the warfarin label based on the influence of the CYP2C9 and VKORC1 genes on anticoagulation-related outcomes. Payers will be facing increasing demand for coverage decisions regarding this technology, but the potential clinical and economic impacts of testing are not clear. OBJECTIVE: To develop a policy model to evaluate the potential outcomes of warfarin pharmacogenomic testing based on the most recently available data. METHODS: A decision-analytic Markov model was developed to assess the addition of genetic testing to anticoagulation clinic standard care for a hypothetical cohort of warfarin patients. The model was based on anticoagulation status (international normalized ratio), a common outcome measure in clinical trials that captures both the benefits and risks of warfarin therapy. Initial estimates of testing effects were derived from a recently completed randomized controlled trial (n = 200). Healthcare cost ($US, year 2007 values) and health-state utility data were obtained from the literature. The perspective was that of a US third-party payer. Probabilistic and one-way sensitivity analyses were performed to explore the range of plausible results. RESULTS: The policy model included thromboembolic events (TEs) and bleeding events and was populated by data from the COUMAGEN trial. The rate of bleeding calculated for standard care approximated bleeding rates found in an independent cohort of warfarin patients. According to our model, pharmacogenomic testing provided an absolute reduction in the incidence of bleeds of 0.17%, but an absolute increase in the incidence of TEs of 0.03%. The improvement in QALYs was small, 0.003, with an increase in total cost of $US162 (year 2007 values). The incremental cost-effectiveness ratio (ICER) ranged from testing dominating to standard care dominating, and the ICER was <$US50,000 per QALY in 46% of simulations. Results were most sensitive to the cost of genotyping and the effect of genotyping. CONCLUSION: Our model, based on initial clinical studies to date, suggests that warfarin pharmacogenomic testing may provide a small clinical benefit with significant uncertainty in economic value. Given the uncertainty in the analysis, further updates will be important as additional clinical data become available. PMID- 20014878 TI - Erlotinib: a pharmacoeconomic review of its use in advanced non-small cell lung cancer. AB - Erlotinib (Tarceva), an oral epidermal growth factor receptor tyrosine kinase inhibitor, is associated with modest improvements in survival in patients with advanced non-small cell lung cancer (NSCLC) who have previously received one or more prior chemotherapy regimens. In a well designed clinical trial in this patient population, median overall survival and progression-free survival were significantly longer in patients receiving erlotinib 150 mg/day than in those receiving placebo. Erlotinib is generally well tolerated, with most adverse events being of mild to moderate severity. A large body of modelled pharmacoeconomic data suggests that second- or third-line erlotinib 150 mg/day is a cost-saving option relative to treatment with the approved second-line intravenous chemotherapies of docetaxel and pemetrexed in patients with advanced NSCLC. In patients who had received at least one prior chemotherapy regimen, erlotinib was predicted to be dominant (i.e. more effective and less costly) or cost saving (i.e. equally effective and less costly) relative to docetaxel or pemetrexed with regard to the cost per QALY or life-year gained in cost effectiveness analyses. Although the effect of erlotinib on overall survival was generally assumed to be equivalent to that of the chemotherapies, the estimated amount of QALYs gained was slightly greater with erlotinib than with docetaxel. In cost-minimization and national budgetary impact analyses, estimated total direct costs with erlotinib were lower than those with docetaxel and pemetrexed, because of the generally lower drug acquisition, administration and adverse event management costs associated with erlotinib. Cost advantages with erlotinib were predicted across analyses, regardless of the type of model developed, specific costs that were included, country that the study was conducted in and year of costing. Sensitivity analyses consistently showed that these results were robust to plausible changes in the key model assumptions. In conclusion, in patients with advanced NSCLC, second- or third-line treatment with erlotinib is clinically effective in improving survival. Available pharmacoeconomic data from several countries, despite some inherent limitations, support the use of erlotinib as a cost-saving treatment relative to chemotherapy with docetaxel or pemetrexed in this patient population. PMID- 20014879 TI - Helicobacter pylori eradication for gastric cancer prevention. PMID- 20014880 TI - World's first delivery of intra-arterial bevacizumab directly into brain tumor of glioblastoma multiforme patients. PMID- 20014881 TI - Decitabine in the treatment of myelodysplastic syndromes. AB - Myelodysplastic syndromes (MDS) are a group of heterogeneous clonal hematopoietic stem cell disorders characterized by ineffective hematopoiesis, peripheral blood cytopenias and a propensity to transform into acute myeloid leukemia. There are few treatment options available for patients with MDS. Studies into the molecular biology of MDS have demonstrated abnormal patterns of DNA methylation that lead to silencing of tumor-suppressor genes. Hypomethylating agents are compounds that have the potential to reverse the aberrant DNA methylation and increase the expression of silenced genes, leading to cellular differentiation and/or apoptosis. Decitabine is a cytidine analogue that has activity as a hypomethylating agent and has been evaluated in the therapy of patients with high risk MDS. Several studies have confirmed the clinical activity of low-dose decitabine in patients with high-risk MDS, leading to responses in approximately 50% of patients, with low treatment-related mortality. Responses have even been seen in patients with high-risk cytogenetic abnormalities, and some studies have demonstrated increased re-expression of genes that were previously silenced by hypermethylation, such as CDKN2B/p15INK4B. There are still some issues concerning the ideal dose and schedule of decitabine for treating patients with MDS. This article focuses on the most recent clinical studies of decitabine for therapy of MDS. PMID- 20014882 TI - Advances in breast cancer treatment: the emerging role of ixabepilone. AB - Although taxanes and anthracyclines have dramatically improved the treatment of breast cancer, resistance to these agents upon continued exposure is almost inevitable. The epothilone ixabepilone was US FDA approved in 2007 based on its demonstrated activity in metastatic breast cancer that is resistant to other approved agents, including taxanes and anthracyclines. Over 2000 patients have now received this agent in clinical trials, clarifying that ixabepilone has efficacy in minimally and heavily pretreated patients and can overcome chemotherapy-induced drug resistance, while maintaining a manageable safety profile. These clinical trials identified a progression-free survival advantage with ixabepilone/capecitabine combination therapy over capecitabine monotherapy. Moreover, certain hard-to-treat subgroups of patients may derive additional benefit from ixabepilone therapy. The objective of this report is to review the updated body of ixabepilone clinical data in breast cancer, as well as key considerations for ixabepilone administration and side-effect management. PMID- 20014883 TI - High-intensity focused ultrasound: where are we and where to from here? AB - High-intensity focused ultrasound (HIFU) has evolved significantly from early work treating cerebral lesions. The ability to treat deep soft-tissue lesions without damaging superficial structures led to it being used for prostate cancer treatment both in the primary and salvage setting. Primary HIFU treatment for prostate cancer leads to 5-year disease free survival rates of up to 70-80% in selected patients with little morbidity; however, comparative studies with established treatment modalities are lacking. Salvage treatment with HIFU leads to significantly more morbidity than primary treatment yet the morbidity appears the same or less than other salvage treatments following external-beam radiation treatment. We believe that with the development of more advanced imaging techniques combined with multimodality prostate imaging that HIFU's future lies in focal treatment of prostate cancer. PMID- 20014884 TI - Image-guided brachytherapy for cervix cancer: from Manchester to Melbourne. AB - Brachytherapy for cervix cancer has evolved rapidly over the last few years through the introduction of image-guided brachytherapy. This has enabled the transition from Manchester-based orthogonal radiograph planning to more conformal treatments, incorporating the concepts of patients' anatomy, tumor characteristics and tumor response with time. The purpose of this article is to outline the evolution of cervix cancer brachytherapy, discuss guidelines and recommendations on image-guided brachytherapy and describe our single-institution conformal image-based brachytherapy experience. PMID- 20014885 TI - First-line treatment of advanced ovarian cancer: current research and perspectives. AB - Epithelial ovarian cancer is the fourth biggest cause of cancer-related death in women. Over recent decades, improvements have been made in treatment outcome in terms of response rate and survival. To date, intensive surgical staging and cytoreduction, followed by primary chemotherapy with the carboplatin-paclitaxel regimen, are considered the gold standard for the management of this disease. Nevertheless, despite good initial response to systemic therapy after optimal debulking surgery, the long-term survival remains poor, with a high risk of recurrence. Furthermore, medical therapy of ovarian cancer impacts quality of life owing to the common occurrence of chemotherapy side effects, such as alopecia, neurotoxicity and fatigue. In order to improve the efficacy and reduce the toxicity of first-line chemotherapy, more than 10,000 women have been involved in worldwide randomized trials in the last 10 years. Several treatment alternatives have been investigated, such as intraperitoneal chemotherapy, alternative doublets and triplet regimens, in the effort to find an optimal first line treatment strategy. In this review we discuss the results of these trials, the recent progresses and the most important ongoing studies, including those with emerging target and biological agents. PMID- 20014886 TI - Update on the sentinel lymph node procedure in vulvar cancer. AB - Currently, standard treatment for early-stage vulvar cancer typically includes wide local excision of the primary tumor and inguinofemoral lymphadenectomy. The morbidity of this treatment is high. The sentinel lymph node (SLN) procedure provides us with a technique for determining the status of the regional lymph nodes with less treatment-related morbidity. Recently, a large multicenter observational study provided level 3 evidence indicating that it appears safe to omit inguinofemoral lymphadenectomy in case of a negative SLN. This review focuses on the different aspects of the SLN procedure in vulvar cancer. PMID- 20014887 TI - Surgical management of chemotherapy-resistant gestational trophoblastic neoplasia. AB - Gestational trophoblastic neoplasia (GTN) are a broad spectrum of placental lesions. Chemotherapy is the primary treatment for GTN and the vast majority of women with GTN are cured with their initial chemotherapy treatment. However, some patients become chemotherapy-resistant and fail to achieve a complete remission following initial chemotherapy and need salvage chemotherapy. A small minority of patients are still unresponsive to salvage multidrug chemotherapy. Currently, adjuvant surgical procedures could be excellent adjuncts to salvage chemotherapy in removing known foci of chemotherapy-resistant disease in selected patients with persistent GTN. This article will review the surgical management of chemotherapy-resistant GTN, focusing on the relevant indication of surgery, factors affecting efficacy and the use of surgical procedures in selected patients. PMID- 20014888 TI - Systemic therapy in recurrent ovarian cancer: current treatment options and new drugs. AB - Most patients with ovarian cancer relapse despite aggressive surgery and platinum taxane-based primary chemotherapy. Further treatment depends on prior response and progression-free interval. Monotherapy is indicated in patients with so called platinum-resistant or -refractory ovarian cancer. The standard treatment for patients with platinum-sensitive recurrent ovarian cancer is platinum-based combination chemotherapy. Cytoreductive surgery is also a treatment option in such patients. Actual treatment options and strategies in recurrent ovarian cancer will also be discussed. Furthermore, this review focuses on new drugs in the treatment of primary and recurrent ovarian cancer. PMID- 20014889 TI - Emerging biological observations in prostate cancer. AB - Emerging biological observations in prostate cancer provide the opportunity for the development of novel approaches to prevention, detection and treatment. Two observations selected for discussion in this review revolve around the mechanisms of action of signaling through the androgen receptor (AR) and the TMPRSS2:ERG chromosomal rearrangement, a fusion protein seen in nearly 50% of prostate cancers. Despite being called androgen-independent, these prostate cancers continue to depend on AR signaling despite low serum androgen levels. AR reactivation in recurrent tumors is hypothesized to occur through multiple mechanisms: AR amplification, AR mutation, active AR signaling (despite low levels of androgen), AR coactivators, ligand-independent AR activation, enhanced local production of androgens, alternative sources of androgen and upregulation in antiapoptotic genes in prostate cancer cells. A major breakthrough in prostate cancer was the identification of recurrent fusions between the androgen-regulated gene, TMPRSS2 and the v-ets erythroblastosis virus E26 oncogene homolog, ERG. This fusion has been identified as a common molecular event in prostate cancer, seen in approximately 50% of primary prostate cancer. It seems clear that this fusion gene plays an early role in prostate cancer development and/or progression, and ongoing work is being performed to elucidate the association between this fusion transcript and cancer aggressiveness. PMID- 20014890 TI - Biomarkers for prostate cancer detection. AB - Since its approval by the US FDA in 1986, prostate-specific antigen (PSA) has been employed to monitor men with a diagnosis of prostate cancer. In 1994, PSA was approved for use in prostate cancer screening and has been employed worldwide. However, due to the limited specificity of PSA for the disease, novel biomarkers are needed for detecting prostate cancer and for determining which cancers need to be treated. This review will discuss the development of new biomarkers for prostate cancer detection and disease prognostication, focusing on recent progress and particular topical issues related to the development and validation of these new markers. PMID- 20014893 TI - Guidance for the classification of carcinogens under the Globally Harmonised System of Classification and Labelling of Chemicals (GHS). AB - The United Nations Conference on Environment and Development (UNCED) has developed criteria for a globally harmonised system of classification and labelling of chemicals (GHS). With regard to carcinogenicity, GHS distinguishes between Category 1 ('known or presumed human carcinogens') and Category 2 ('suspected human carcinogens'). Category 1 carcinogens are divided into Category 1A ('known to have carcinogenic potential for humans'), based largely on human evidence, and 1B ('presumed to have carcinogenic potential for humans'), based largely on experimental animal data. Concerns have been raised that the criteria for applying these carcinogenicity classifications are not sufficiently well defined and potentially allow different conclusions to be drawn. The current document describes an attempt to reduce the potential for diverse conclusions resulting from the GHS classification system through the application of a series of questions during the evaluation of data from experiments with rodents; epidemiological data, which could lead to Category 1A, have not been considered. Answers to each question can lead either to a classification decision or to the next question, but this process should only be implemented in an environment of informed scientific opinion. The scheme is illustrated with five case studies. These questions are: (1) Has a relevant form of the substance been tested? (2) Is the study design relevant to human exposure? (3) Is there a substance-related response? (4) Is the target tissue exposure relevant to humans? (5) Can a mode of action be established? (6) Is the mode of action relevant to humans? (7) What is the potency? PMID- 20014894 TI - Inhibition studies of soybean (Glycine max) urease with heavy metals, sodium salts of mineral acids, boric acid, and boronic acids. AB - Various inhibitors were tested for their inhibitory effects on soybean urease. The K(i) values for boric acid, 4-bromophenylboronic acid, butylboronic acid, and phenylboronic acid were 0.20 +/- 0.05 mM, 0.22 +/- 0.04 mM, 1.50 +/- 0.10 mM, and 2.00 +/- 0.11 mM, respectively. The inhibition was competitive type with boric acid and boronic acids. Heavy metal ions including Ag(+), Hg(2+), and Cu(2+) showed strong inhibition on soybean urease, with the silver ion being a potent inhibitor (IC(50) = 2.3 x 10(-8) mM). Time-dependent inhibition studies exhibited biphasic kinetics with all heavy metal ions. Furthermore, inhibition studies with sodium salts of mineral acids (NaF, NaCl, NaNO(3), and Na(2)SO(4)) showed that only F(-) inhibited soybean urease significantly (IC(50) = 2.9 mM). Competitive type of inhibition was observed for this anion with a K(i) value of 1.30 mM. PMID- 20014895 TI - Sulfonamides and trimethoprim. PMID- 20014897 TI - A simplified three-times weekly daptomycin dosing regimen for chronic hemodialysis patients. AB - Approximately 2.3 million patients worldwide are undergoing chronic renal replacement therapy. In that population, acute infections substantially contribute to the excessive morbidity and mortality. The risk for invasive methicillin-resistant Staphylococcus aureus infections in this population is approximately 100-fold higher than in the general population, therefore dialysis patients currently account for up to approximately 15% of all invasive methicillin-resistant Staphylococcus aureus infections. A simplified three-times weekly dosing regimen for hemodialysis patients now allows for practical, hassle free and effective treatment with daptomycin, which is licensed for the treatment of complicated skin and soft tissue infections, including resistant strains of Staphylococcus aureus and life-threatening Gram-positive infections, vancomycin resistant enterococcal infections and right-sided endocarditis. PMID- 20014898 TI - Community-acquired upper respiratory tract infections and the role of third generation oral cephalosporins. AB - Common community-acquired infections include those of the upper respiratory tract. In the 1990s, the antimicrobial treatment of upper respiratory tract infections focused on penicillin-resistant Streptococcus pneumoniae. However, following the introduction of a pneumococcal conjugate vaccine, a decrease in invasive pneumococcal disease occurred, and in the case of otitis media a shift towards Haemophilus influenzae as the predominant causative pathogen was observed. Future antimicrobial therapy for outpatient upper respiratory tract infections may need to focus on pathogens such as penicillin-susceptible S. pneumoniae, beta-lactamase-producing H. influenzae, beta-lactamase-negative amoxicillin-resistant H. influenzae and Moraxella catarrhalis. In these circumstances, third-generation oral cephalosporins, such as cefixime and cefdinir, could be increasingly used as an optional first-line therapy in community practice for upper respiratory tract infections suspected to be caused by these key pathogens, as an alternative to amoxicillin-clavulanate. PMID- 20014899 TI - Male circumcision for HIV prevention: developments from sub-Saharan Africa. AB - The demonstration of a strong effect of male circumcision on reducing HIV acquisition has provided impetus for this intervention to be adopted more widely in areas of the world where HIV prevalence is high and rates of male circumcision are low. This perspective reviews recent research findings and provides a summary of progress in various countries. PMID- 20014900 TI - A clear and present danger: tick-borne diseases in Europe. AB - Ticks can transmit a variety of viruses, bacteria or parasites that can cause serious infections or conditions in humans and animals. While tick-borne diseases are becoming an increasing and serious problem in Europe, tick-borne diseases are also responsible for major depressions in livestock production and mortality in sub-Saharan Africa, Latin America and Asia. This review will focus on the most important circulating tick-transmitted pathogens in Europe (Borrelia spp., Anaplasma phagocytophilum, Babesia spp., tick-borne encephalitis virus, Rickettsia spp. and Crimean-Congo hemorrhagic fever virus). PMID- 20014901 TI - Is antibiotic prophylaxis effective in preventing urinary tract infections in patients with vesicoureteral reflux? AB - The main concern in the management of children with vesicoureteral reflux (VUR) is the prevention of urinary tract infections and avoidance of renal damage. Medical management has been recommended or preferentially suggested over surgery in all but a few select clinical situations. Prophylactic antibiotics are prescribed routinely in the management of young children with radiological evidence of VUR following an episode of acute pyelonephritis. Prophylaxis is generally maintained until the VUR resolves spontaneously or is corrected surgically. Although the administration of prophylactic antibiotics has been universal in children with VUR, some authors have reported that long-term antibiotic prophylaxis does not fully prevent urinary tract infections or scarring, that antibiotic-related adverse events are known to occur, and that the incidence of pyelonephritis does not increase in spite of prophylactic antibiotic cessation. Recently, four prospective, randomized, controlled trials of antibiotic prophylaxis for preventing pyelonephritis and renal scarring were reported and some placebo-controlled, double-blind prospective studies are ongoing. The goal of this review is to evaluate the treatment of VUR using antibiotic prophylaxis, and its advantages and disadvantages based on appropriate descriptions and studies in the literature. PMID- 20014902 TI - Geographic map and evolution of primary Helicobacter pylori resistance to antibacterial agents. AB - Antibiotic resistance in Helicobacter pylori is the major cause of eradication failure. Primary H. pylori susceptibility patterns, however, are becoming less predictable. Currently, high (> or =20%) clarithromycin resistance rates have been observed in the USA and in developed countries in Europe and Asia, while the highest (> or =80%) metronidazole-resistance rates have been reported in Africa, Asia and South America. Primary quinolone-resistance rates of 10% or more have already been reported in developed countries in Europe and Asia. Primary amoxicillin resistance has been low (0 to <2%) in Europe but higher (6-59%) in Africa, Asia and South America. Similarly, tetracycline resistance has been absent or low (<5%) in most countries and higher (9-27%) in Asia and South America. The increasing clarithromycin and quinolone resistance, and multidrug resistance detected in 0 to less than 5% in Europe and more often (14.2%) in Brazil are worrying. Growing resistance often parallels national antibiotic consumption and may vary within patient groups according to the geographic region, patient's age and sex, type of disease, birthplace, other infections and other factors. The geographic map and evolution of primary H. pylori resistance are clinically important, should be considered when choosing eradication regimens, and should be monitored constantly at national and global levels in an attempt to reach the recently recommended goal of eradication of more than 95%. PMID- 20014903 TI - Multidrug-resistant Pseudomonas aeruginosa and Acinetobacter baumannii: resistance mechanisms and implications for therapy. AB - Pseudomonas aeruginosa and Acinetobacter baumannii are major nosocomial pathogens worldwide. Both are intrinsically resistant to many drugs and are able to become resistant to virtually any antimicrobial agent. An increasing prevalence of infections caused by multidrug-resistant (MDR) isolates has been reported in many countries. The resistance mechanisms of P. aeruginosa and A. baumannii include the production of beta-lactamases, efflux pumps, and target-site or outer membrane modifications. Resistance to multiple drugs is usually the result of the combination of different mechanisms in a single isolate or the action of a single potent resistance mechanism. There are many challenges in the treatment of MDR P. aeruginosa and A. baumannii, especially considering the absence of new antimicrobials in the drug-development pipeline. In this review, we present the major resistance mechanisms of P. aeruginosa and A. baumannii, and discuss how they can affect antimicrobial therapy, considering recent clinical, microbiological, pharmacokinetic and pharmacodynamic findings of the main drugs used to treat MDR isolates. PMID- 20014906 TI - Stereotactic neurosurgical treatment of drug addiction. AB - BACKGROUND: Stereotactic neurosurgery is a well-developed and accepted treatment for movement disorders like Parkinson's disease. OBJECTIVE: Review use of this neurosurgical intervention to treat drug addiction since the 1960s. METHODS: Literature review of case and series reports. RESULTS: Stereotactic neurosurgery has shown promising results in reducing drug addiction, but it has also caused severe side effects. Thus, surgeons have sought to refine the stereotactic lesion procedure and its target over the past 50 years. CONCLUSIONS AND SIGNIFICANCE: Recent developments in stereotactic technique and deep brain stimulation (DBS) may make stereotactic neurosurgery a viable and reversible treatment for drug addiction. PMID- 20014904 TI - Use of vancomycin pharmacokinetic-pharmacodynamic properties in the treatment of MRSA infections. AB - Vancomycin is a commonly used antimicrobial in patients with methicillin resistant Staphylococcus aureus (MRSA) infections. Increasing vancomycin MIC values in MRSA clinical isolates makes the optimization of vancomycin dosing pivotal to its continued use. Unfortunately, limited data exist regarding the optimal pharmacokinetic-pharmacodynamic (PK-PD) goal to improve bacterial killing and clinical outcomes with vancomycin. The hallmark study in this area suggests that achieving an AUC to MIC ratio of over 400 improves the likelihood of achieving these outcomes. Challenges in the implementation of PK-PD-based dosing for vancomycin include current methodologies utilized in microbiology laboratories, as well as intra- and interpatient pharmacokinetic variability. Individualized dosing based on MIC and specific patient factors is important to achieve optimal outcomes from vancomycin therapy. PMID- 20014907 TI - Multiple risk factor model predicting cannabis use and use disorders: a longitudinal study. AB - BACKGROUND: Identification of factors associated with the initiation and continuation of cannabis use is important for any preventive work. OBJECTIVES: This study aimed to examine the early life course predictors of cannabis use and the development of cannabis use disorder in early adulthood. METHODS: Data from Mater Hospital and University of Queensland Study of Pregnancy (MUSP), a population-based prospective birth cohort study. Participants were a cohort of 2,493 young adults who completed the life-time version of the Composite International Diagnostic Interview-computerized version (CIDI-Auto) at the 21 year follow-up, and for whom data were available from previous follow-ups. Ever use and age at first use of cannabis was assessed via self-report, and cannabis use disorder was measured based on Diagnostic and Statistical Manual version IV (DSM-IV) diagnostic criteria. Life course predictors were obtained between the child's birth and the 14-year follow-up. RESULTS: Of 2,493 young adults, 51.4% reported having ever used cannabis, and 21.1% of young adults were classified as having ever had a cannabis use disorder. Child's gender, changes in maternal marital status, maternal smoking, child school performance, childhood sexual abuse, early adolescence smoking and alcohol consumption, and adolescent aggression/delinquency were strongly associated with young adult cannabis use and use disorder. Exposure to multiple risk factors was associated with greater risk of outcomes. CONCLUSIONS: There are a number of strong predictors of cannabis use identified in this study. They suggest that the social context within which children are reared has a major influence on cannabis use and use disorders. PMID- 20014908 TI - A study on Fu-Yuan Pellet, a traditional chinese medicine formula for detoxification of heroin addictions. AB - BACKGROUND: Efforts toward researching effective and safe therapies for the treatment of drug addiction and acute heroin withdrawal syndrome remain important objectives in the field of drug addiction. Traditional Chinese medicine (TCM) is viewed as a potential approach to the treatment of drug addiction, and especially to opiate addiction. OBJECTIVES: The objective is to investigate the efficacy and safety of Fu-Yuan Pellet (FYP), a Chinese traditional medicine formula, for the treatment of acute heroin withdrawal syndrome. METHODS: A multicenter, randomized, double-blind, double-dummy, and positive-controlled trial was conducted at 3 drug abuse treatment centers in China. Patients (n = 225) who met a diagnosis of opiate dependence based on DSM IV classification were recruited for this study, ranging in age from 18 to 55 years. Inclusion criteria included a heroin-positive urinalysis, as measured between 8 to 36 hours from last use of heroin, and total withdrawal syndrome scores above 50 before treatment (actual range 65-140). These patients were treated with either FYP or lofexidine in a fixed schedule of doses for 10 days. The total withdrawal syndrome scores and the daily reduction rate were used to measure the effect of FYP vs. lofexidine. RESULTS: Both treatments significantly reduced withdrawal symptoms by day 3, but there was no significant difference overall between lofexidine and FYP in efficacy or safety. CONCLUSIONS AND SCIENTIFIC SIGNIFICANCE: This clinical trial has shown that FYP is effective in the treatment of moderate-to-severe acute heroin withdrawal and has few adverse effects compared to lofexidine. Further study is warranted to determine whether FYP is similar to lofexidine in its potential for reducing stress induced opiate relapse. PMID- 20014909 TI - Atomoxetine attenuates dextroamphetamine effects in humans. AB - BACKGROUND: Although preclinical studies support the contribution of the noradrenergic system activation in mediating the acute effects of amphetamines, these findings have not been followed up in clinical studies. OBJECTIVES: To examine the effects of atomoxetine, a norepinephrine transporter inhibitor, on subjective, physiological, and plasma cortisol responses to dextroamphetamine in 10 healthy volunteers. METHODS: Subjects were randomly assigned to a sequence of atomoxetine (40 mg/day) or placebo treatments each lasting for 4 days. On Day 4 of each treatment period, responses to a single 20 mg/70 kg dose of dextroamphetamine were assessed. RESULTS: Atomoxetine treatment attenuated dextroamphetamine-induced increases in systolic and diastolic blood pressure and plasma cortisol as well as the self-report ratings of "stimulated," "high," and "good drug effects." CONCLUSIONS: These findings are consistent with previous preclinical studies supporting the role of the noradrenergic system in mediating acute amphetamine responses. SCIENTIFIC SIGNIFICANCE: Atomoxetine's capacity to attenuate some of the physiological and subjective responses to dextroamphetamine supports its potential use for stimulant addiction. PMID- 20014910 TI - Substance abuser impulsivity decreases with a nine-month stay in a therapeutic community. AB - BACKGROUND: Substance abuse continues to be a major public health problem. Keeping substance abusers in treatment is a challenge, and researchers continue to investigate ways to increase retention. OBJECTIVE: The aim of this study was to investigate the relationship between impulsivity in substance abusers and length of stay in the context of therapeutic community. METHODS: The Barratt Impulsiveness Scale- 11 (BIS-11) was used to assess impulsivity in 138 substance abusers at admission and at nine months in a therapeutic community. RESULTS: Impulsivity significantly decreased in subjects who completed nine months in the therapeutic community. Legal stipulation increased length of stay, on average, by three months. On admission, female participants were on average more impulsive than their male counterparts. CONCLUSION: Impulsivity decreased in subjects who remained in therapeutic community for nine months although self-reported impulsivity at baseline did not seem to be associated with retention. SCIENTIFIC SIGNIFICANCE: Therapeutic community factors contribute to a decrease in self reported impulsivity and these factors might be enhanced to increase retention in therapeutic community. PMID- 20014911 TI - Relationships between drinking onset, alcohol use intensity, and nighttime risk behaviors in a college bar district. AB - OBJECTIVE: To identify antecedents of risk behavior events in college bar patrons. METHODS: In this nighttime field study, self-report data and alcohol intoxication readings were collected from patrons immediately upon exiting bars (n = 618). RESULTS: Multilevel structural equation modeling revealed positive associations between age of drinking onset and both alcohol use intensity in the past year and recent bar-going frequency. In turn, alcohol use intensity in the past year was positively associated with bar-going frequency and intoxication at bar exit. An association between drinking onset and bar-going frequency was mediated by alcohol use intensity in the past year. CONCLUSIONS: Discernable paths from age of drinking onset to monthly bar-going frequency and intoxication level after leaving a bar can be identified. The results highlight the critical role of drinking onset in development of college student alcohol abuse. Research is needed to determine whether college bars are environmental pathogens mediating between genetic risk factors and patron risk behavior. PMID- 20014912 TI - Characteristics of opioid-using pregnant women who accept or refuse participation in a clinical trial: screening results from the MOTHER study. AB - BACKGROUND: Although concerns arise about the generalizability of results from Randomized Controlled Trials (RCTs), few studies systematically examine this issue. OBJECTIVES: This study compared the characteristics of 427 opioid-using pregnant women who did (n = 208) and did not consent (n = 219) to enrollment in a multicenter clinical trial of agonist medications (i.e., the MOTHER study). METHODS: Logistic regression models were used to compare consenters and non consenters to examine the effect of screening variables on the likelihood of consenting. RESULTS: Of nine characteristics examined, most differences did not reach statistical significance. Consenting participants were less likely than non consenting women to be currently enrolled in a methadone maintenance program (74.5% vs. 84.5%, p =.01). CONCLUSION AND SCIENTIFIC SIGNIFICANCE: These data show that the recruited sample of drug-dependent pregnant women enrolled in an intensive RCT is representative of the larger population of treated opioid dependent patients and supports the generalizability of randomized controlled trials in this population. PMID- 20014913 TI - D-cycloserine and cocaine cue reactivity: preliminary findings. AB - BACKGROUND: D-cycloserine (DCS), a partial glutamate N-methyl-D-aspartic acid (NMDA) receptor agonist, enhances extinction of conditioned fear responding in rodents and facilitates exposure-based learning in humans with anxiety disorders. OBJECTIVES: This preliminary study investigates DCS pretreatment on response to cocaine cues in cocaine-dependent subjects. METHODS: Ten cocaine-dependent subjects were randomly assigned to receive either 50 mg DCS or matching placebo two hours before each of two 1-hour cocaine cue exposure sessions one day apart. HR and craving ratings were obtained before and during cue exposure sessions. RESULTS: There was a trend towards increased craving to cocaine cues in cocaine dependent individuals after administration of DCS. CONCLUSIONS: The administration of DCS prior to cue exposure sessions may facilitate response activation. SCIENTIFIC SIGNIFICANCE: While facilitation of extinction-based learning by DCS may have therapeutic potential for cocaine dependence, this drug may exhibit a different profile in cocaine-dependent individuals as compared to those with anxiety disorders. PMID- 20014914 TI - Comparison of tuberculin skin testing reactivity in opioid-dependent patients seeking treatment with methadone versus buprenorphine: policy implications for tuberculosis screening. AB - BACKGROUND: Buprenorphine's availability in primary care settings offers increased access to treatment and linkage to primary care for opioid-dependent patients. Currently, tuberculin skin testing (TST) is recommended for patients enrolling in methadone maintenance treatment (MMT), but not for those enrolling in buprenorphine maintenance treatment (BMT). OBJECTIVES: To compare TST screening results in enrollees in BMT and MMT programs and assess the correlates of TST positivity among these subjects. METHODS: A cross-sectional analysis of a retrospective cohort study was conducted to compare concurrent TST results among contemporaneously matched groups of MMT and BMT patients in the same community. RESULTS: TST positivity was approximately 9% in both MMT and BMT settings (p = .27). Increased TST positivity was associated with being Black (AOR = 3.53, CI = 1.28-9.77), Hispanic (AOR = 3.11, CI = 1.12-8.60), and having higher education (AOR = 3.01, CI = 1.20-7.53). CONCLUSIONS: These results confirm a similar high prevalence of TST positivity in opioid-dependent patients enrolling in MMT and BMT programs. Racial and ethnic health disparities remain associated with TST positivity, yet a relationship between higher education and tuberculosis requires further investigation. SCIENTIFIC SIGNIFICANCE: These data suggest the importance of incorporating TST screening in emerging BMT programs as a mechanism to provide increased detection and treatment of tuberculosis infection in opioid-dependent patient populations. PMID- 20014915 TI - Reported reasons for initiating drug use among drug-dependent adolescents and youths in Yunnan, China. AB - BACKGROUND: As an entry point for illicit drug trafficking, Yunnan was hit hard by the twin epidemics of drug abuse and HIV/AIDS. There are no empirical studies regarding the reasons for initiation of drug use in Yunnan. OBJECTIVES: The study investigated the reasons for initiating drug use in drug-dependent adolescents and youths in five regions in Yunnan. METHODS: The data were collected in 2004 2005 from a stratified random sample of 467 registered drug addicts under age 25. Pearson chi-square tests were used to analyze the association of reasons for drug use with sociodemographic factors, knowledge of drug related harm, and access to illicit drugs. RESULTS: Curiosity was the most commonly reported reason for initiating drug use and the majority of the respondents reported having little or no knowledge of drug-related harm. The majority of the drug abusers initiated drug use at the age of 13-14 years or younger. CONCLUSIONS: There is a potential for sustained educational programs to reduce the high prevalence of adolescent drug use in Yunnan. SCIENTIFIC SIGNIFICANCE: This is the first empirical study that has investigated reasons for initiating drug use based on a random sample of registered drug users in a high risk province of China. PMID- 20014916 TI - Healthcare chief executive officers: the best of times; the worst of times. PMID- 20014917 TI - Regulatory polymorphisms in key candidate genes for disease susceptibility and drug response: a mandate for valid genetic biomarkers. PMID- 20014919 TI - Diagnosis of uncertain primary tumors with the Pathwork tissue-of-origin test. AB - Clinical workup of metastatic malignancies of unknown origin is an arduous and expensive process, which is reported to be unsuccessful in up to 30% of cases. Global gene expression-based molecular testing may offer accurate classification of metastatic tumors in which a primary site has not been identified. Recently, the US FDA cleared the Pathwork((R)) tissue-of-origin test, which is a gene expression microarray-based test that quantifies the molecular similarity of tumor specimens to 15 known tissue types. A blinded, multicenter validation on poorly differentiated and undifferentiated tumors showed 87.8% sensitivity and 99.4% specificity in frozen tissue samples. The availability of ancillary gene expression-based molecular tests for tissue of origin determination represents a milestone in cancer patient management as part of the personalized medicine revolution. PMID- 20014920 TI - Geographic ancestry and markers of preterm birth. AB - Several biomarkers associated with spontaneous preterm birth (PTB) have been discovered over the last decade. Many of these markers, such as cytokines, are associated with infection and inflammation. As such, these biomarkers represent biologically plausible candidates for assessing those at risk of PTB. However, in the early association studies of biomarker-pregnancy outcome, the geographic ancestry of subjects was not considered. Based on more recent data, it is becoming increasingly evident that these biomarkers, and a universal approach that uses a single biomarker, fail to provide adequate assessment of risk in all subjects. Rather, recent data support the conclusion that some markers associate in subjects of African descent and another nonoverlapping set associates in subjects of European descent. These data indicate that diagnostic or predictive tests will have to use different biomarkers for different sets of subjects. If this is true, it poses severe restrictions on how to predict outcome or perform tests of association, and may make it impossible to determine risk or provide proper intervention. An alternative is presented that, although not yet proven, may make it possible to use a common set of biomarkers and their relationships to assess risk. PMID- 20014921 TI - Novel clinico-genome network modeling for revolutionizing genotype-phenotype based personalized cancer care. AB - Although cancer heterogeneity, even within individual tumors with different treatment responses of subcloncal cells populations, suggests the need for personalized medicine, most funding and efforts go to conventional single gene based research and comparative-effectiveness research. Cancer arises from changes in the DNA sequence in the genomes of cancer cells. These accelerating somatic mutations dysregulate signaling pathways, including EGFR, Wnt/Notch, Hedgehog and others, with a central role in cell growth, proliferation, survival, angiogenesis and metastasis. All of these genetic alterations can now be discovered using next generation DNA sequencing technology. This high-throughput technology can achieve two major goals: first, to complete the catalogue of driver mutations, including point mutations, rearrangements and copy-number changes, by full and targeted sequencing; and second, to explore the functional role of cancer genes and their interactions by genome-wide RNA, serial analysis of gene expression, microRNAs, protein-DNA interactions, and comprehensive analyses of transcriptomes and interactomes. This review article discusses the challenges, including costs, in completing the catalogue of driver mutations for each cancer type and understanding how cancer genomes operate as whole biological systems. Now high quality clinical treatment and outcomes (death or survival) data from biobanks, and extensive genetics and genomics data for some common tumors, including breast, colorectal and pancreatic cancer, are available. In this article, we will describe how all these clinical and genetics data could be integrated into reverse engineering-based network modeling to approach the extremely complex genotype-phenotype map. This clinico-genome systems model, published for the first time, opens the way for the discovery of new molecular innovations, both predictive markers and therapies, towards personalized treatment of cancer. Instead of the comparative-effectiveness research or personalized medicine debate, harmonization of both can revolutionize cancer management. PMID- 20014922 TI - Discovery of prostate cancer biomarkers by microarray gene expression profiling. AB - Prostate cancer is the most commonly diagnosed malignancy in males in the Western world. This review focuses on advances in biomarker discovery for prostate cancer by microarray profiling of mRNA and microRNA expression. Novel biomarkers are strongly needed to enable more accurate detection of prostate cancer, improve prediction of tumor aggressiveness and facilitate discovery of new therapeutic targets for tailored medicine. Promising molecular markers identified from gene expression profiling studies include AMACR, EZH2, TMPRSS2-ERG, miR-221 and miR 141, which are described in more detail. In addition, a compilation of prognostic gene expression signatures for prediction of prostate cancer patient outcome is provided, and their possible clinical utility is discussed. Furthermore, limitations in the application of microarray-based expression profiling for identification of prostate cancer biomarkers are addressed. PMID- 20014923 TI - Putative molecular signatures for the imaging of prostate cancer. AB - Prostate cancer is the most common cancer among men worldwide and is the second leading cause of death among those over 50 years of age in the USA. However, many men who develop a prostate tumor never exhibit symptoms in the early stage of the disease or even before it spreads to other parts of the body, such as bones and lymph nodes. Therefore, the successful prevention and treatment of prostate cancer relies on the early and accurate detection of the disease. Although prostate-specific antigen has been extensively used as a serum biomarker to detect prostate tumors in the past 20 years, this screening method has suffered from a lack of specificities and sensitivities, despite its wide use. Furthermore, fluorine-18-labeled fluorodeoxyglucose, a radiopharmaceutical useful in the detection (using PET) of various solid tumors, is not accurate for imaging cancer of the prostate. Therefore, there is an underlying necessity to discover upregulated tumor-specific markers that may serve as molecular targets for the imaging of prostate cancer. This review summarizes the most recent advances made in the discovery of tumor-specific signatures that could be useful for imaging and accurate detection of prostate cancer, using the tools of bioinformatics, genomics, proteomics and metabolomics approaches. Also introduced is the recent development of a few promising techniques, such as functional MRI, to facilitate the detection of prostate tumor signatures. PMID- 20014924 TI - Prospects for personalized medicine with inhibitors targeting the RAS and PI3K pathways. AB - Tumor genetic analyses have revealed that the signaling pathways regulated by PI3K and RAS are of fundamental importance in a wide variety of human neoplasms, leading to intensive efforts to develop therapeutics that block signaling through these two key pathways. Both pathways frequently undergo a variety of activating alterations, including oncogenic mutations, amplification events and loss of tumor-suppressor genes that are thought to confer aggressive growth properties and enhance survival on neoplastic cells. An attractive hypothesis is that these alterations provide an indication that a particular tumor is addicted to signaling through the affected pathway, thus may provide ideal candidate predictive biomarkers to target these inhibitors to appropriate patient populations. This review highlights recent preclinical progress made on understanding the predictive value of key pathway alterations in response to targeted therapeutics directed against PI3K, AKT, mTOR, BRAF and MEK, and the prospects for biomarker-driven clinical strategies for such inhibitors. PMID- 20014925 TI - Should microchimerism turn into rejection prophylactics? AB - Non-self cells can circulate in the body of an individual after any sort of contact with an allogeneic source of cells, thus creating a situation of chimerism that can be transient or prolonged over time. This situation may appear after stem cell transplantation, pregnancy, transfusion or transplantation. Concerning transplantation, many hypotheses have been formulated regarding the existence, persistence and role of these circulating cells in the host. We will review the principal hypotheses that have been formulated for years since the first description of non-self circulating cells in mammals to the utilization of artificially induced chimerism protocols for the achievement of tolerance. PMID- 20014929 TI - Future directions in stenting. PMID- 20014930 TI - Corporate influence over planning and presentation of clinical trials: beauty and the beast. PMID- 20014931 TI - Genetic risk factors for venous thrombosis: the end of the beginning? PMID- 20014934 TI - Cabergoline therapy for prolactinomas: is valvular heart disease a real safety concern? AB - Dopamine agonists (DAs) are the first-line therapy for the treatment of hyperprolactinemia, with cabergoline, an ergot-derived selective D(2)-receptor agonist, being the preferred and most widely used drug. Recent studies reported cardiac valve regurgitations in patients with Parkinson's disease treated with high doses of DA, raising concerns about the safety of cabergoline in patients with hyperprolactinemia. To date, seven case-control studies have examined the potential association between cardiac valvular abnormalities and cabergoline therapy in patients with hyperprolactinemia. Overall, a total of 463 patients exposed to low doses of cabergoline (mean cumulative doses: 204-443 mg) for a mean duration of 45-79 months have been included in these studies. Patients in all the studies were asymptomatic without clinical signs of cardiac disease. Six studies did not show any association between cabergoline therapy and clinically relevant valvular regurgitation, whereas one study found an increased rate of moderate tricuspid regurgitation. In this report, we review and discuss the results of these studies and emphasize the limitations of the methodology used in the published literature. The clinical significance of the present findings has yet to be confirmed by future larger prospective studies with rigorous echocardiographic protocols and prolonged duration of follow-up. PMID- 20014933 TI - Cardiovascular MRI in small animals. AB - Imaging studies of cardiovascular disease in small rodents have become a prerequisite in preclinical cardiovascular research. Transgenic and gene-knockout models of cardiovascular diseases enables the investigation of the influence of single genes or groups of genes on disease pathogenesis. In addition, experimental and genetically altered models provide valuable in vivo platforms to investigate the efficacy of novel drugs and contrast agents. Owing to the excellent soft tissue contrast, high spatial and temporal resolution, as well as the tomographic nature of MRI, anatomy and function can be assessed with unique accuracy and reproducibility. Furthermore, using novel targeted MRI contrast agents, molecular changes associated with cardiovascular disease can be investigated in the same imaging session. This review focuses on recent advances in hardware, imaging sequences and probe design. PMID- 20014935 TI - Myocardial viability imaging and revascularization in chronic ischemic left ventricular systolic dysfunction. AB - The high rate of periprocedural morbidity and mortality associated with revascularization in moderate-to-severe left ventricular systolic dysfunction has provided the rationale for noninvasive viability imaging. The most established viability imaging techniques are PET, single-photon emission computed tomography and dobutamine echocardiography. Cardiac MRI is gaining widespread use and accessibility, and computed tomography is emerging as a promising technique. Each imaging modality has unique advantages but also suffers from limitations. Furthermore, evidence of the impact of viability imaging on patient outcomes has generally been limited to observational studies. Uncertainty remains regarding the optimal treatment strategy in this patient subset. This review describes the current status of viability imaging and revascularization in chronic moderate-to severe ischemic left ventricular systolic dysfunction. PMID- 20014936 TI - Left atrial appendage: structure, function, imaging modalities and therapeutic options. AB - The left atrial appendage (LAA) is a common source of cardiac thrombus formation and systemic embolism. It is a 'blind' cul-de-sac and multilobed anatomic structure with variable anatomy. Therefore, it requires detailed evaluation in multiple imaging planes to evaluate for thrombus formation. Transesophageal echocardiography is the most common imaging modality used to rule out LAA thrombus. Doppler imaging enhances understanding of LAA function. 3D imaging of the LAA with live 3D transesophageal echocardiography, computed tomography and MRI may be further utilized for thrombus detection, as well as for sizing, and the development of new transcatheter occluder devices for LAA to prevent thrombus formation is needed. PMID- 20014937 TI - Cardiac magnetic resonance in hypertrophic cardiomyopathy: current state of the art. AB - Hypertrophic cardiomyopathy is a complex disorder with significant heterogeneity in clinical characteristics and natural history. Traditionally, the diagnosis has been based on clinical assessment and echocardiography; however, persistent challenges in its noninvasive evaluation remain. Hence, improved diagnostic techniques could lead to better risk stratification of patients, which would potentially identify patients likely to benefit from effective therapies. Recent studies have demonstrated the increasing utility of cardiac magnetic resonance in the management of this disease. With the increasing utilization of genetics, cardiac magnetic resonance is likely to play an even more important role in discerning the subtle morphologic differences seen in such patients with similar genotypic profiles. PMID- 20014938 TI - Screening for heart disease: C-reactive protein versus coronary artery calcium. AB - The Framingham model has become the standard to assess future risk of coronary artery disease. Several nontraditional risk factors have been suggested to improve risk stratification to provide a new algorithm for predicting coronary heart disease (CHD). Two factors with the most available data include C-reactive protein (CRP) and coronary artery calcification (CAC). CRP, a well-established biomarker of inflammation, has shown very inconsistent data to predict CHD, and no study to date shows improvement in the C-statistic; in addition, reclassification is modest (<10% of patients are reclassified to higher or lower risk categories). CAC seems to hold a greater promise in the prediction of CHD and demonstrates a stronger relationship to future cardiac events, with all studies demonstrating improvement in the C-statistic when added to the Framingham risk score. Measurement of CAC consistently provides reclassification of patients more accurately to the Framingham risk model. Only four studies have evaluated both CAC and CRP in the same cohort for future cardiovascular events, but all of these showed significant prediction for coronary artery disease using CAC and no significant prediction ability for CRP. Current review of the literature available suggests CAC to be more relevant in evaluating CHD in a clinical setting. Given the higher cost of CAC scanning compared with CRP, cost effectiveness studies are still needed. PMID- 20014939 TI - The evolution of reciprocity: social types or social incentives? AB - The vast majority of human beings regularly engage in reciprocal cooperation with nonrelated conspecifics, and yet the current evolutionary understanding of these behaviors is insufficient. Intuitively, reciprocity should evolve if past behavior conveys information about future behavior. But it is not straightforward to understand why this should be an outcome of evolution. Most evolutionary models assume that individuals' past behavior informs others about their stable social type (defector, cooperator, reciprocator, etc.), which makes it sensible to reciprocate. In this article, after describing the central source of difficulty in the evolutionary understanding of reciprocity, I put forward an alternative explanation based on a work by O. Leimar. It consists of taking into account the fact that the payoffs to individuals in social interactions can change through time. This offers a solution because individuals' past behavior then signals their payoffs, which also makes it sensible to reciprocate. Even though the overwhelming majority of evolutionary models implicitly endorse the social types mechanism, I argue that the social incentives mechanism may underlie reciprocity in humans. PMID- 20014940 TI - Synthetic, non-natural analogs of ceramide elevate cellular ceramide, inducing apoptotic death to prostate cancer cells and eradicating tumors in mice. AB - The anticancer effects of synthetic, non-natural analogs of ceramide were tested using human TSU-Pr1 prostate cancer cells in-vitro as well as in-vivo, following their effects on tumors development in mice. When incubated with the cultured cancer cells, the analogs elevated cellular ceramide and induced a cytotoxicity and death by apoptosis. When a ceramide analog was injected intradermally or intraperitoneally into BALB/c-Nude or NOD-SCID mice bearing a human prostate tumor, a considerable regression of the tumor was observed. The synthetic ceramide analogs should thus be further investigated as potential anticancer drugs. PMID- 20014941 TI - Oligonucleotide DNA microarray profiling of lung adenocarcinoma revealed significant downregulation and deletions of vasoactive intestinal peptide receptor 1. AB - The purpose of this study was to find novel gene(s) involved in the development of lung adenocarcinoma (AD). Using DNA microarrays, we identified 31 up-regulated and 8 downregulated genes in 12 AD. Real time PCR was used to measure expression of VIPR1 and SPP1 mRNA and possible losses or gains of genes in 32 AD. We describe significant upregulation of the SPP1 gene, downregulation of VIPR1, and losses of the VIPR1 gene. Our findings complement a proposed VIPR1 tumor suppressor role, in which deletions in the 3p22 chromosome region are an important mechanism leading to loss of the VIPR1 gene. PMID- 20014942 TI - Downregulation of N-cadherin expression inhibits invasiveness, arrests cell cycle and induces cell apoptosis in esophageal squamous cell carcinoma. AB - Some studies have demonstrated that N-cadherin is upregulated in more invasive cancer cell lines and tumors and plays a key role in intercellular adhesion. However, the understanding on the roles N-cadherin plays in esophageal squamous cell carcinoma (ESCC) is still poor. Our data showed that knock-down of N cadherin in ESCC cell line (EC9706) could arrest cell cycle at G0/G1 phase, induce cell apoptosis, reduce the invasiveness in vitro, and inhibit the tumor formation in vivo. These results suggest that N-cadherin is an important factor in the progression and metastasis of ESCC and N-cadherin may serve as a potential molecular target for biotherapy of ESCC. PMID- 20014943 TI - Expression of WISP3 and RhoC genes at mRNA and protein levels in inflammatory and noninflammatory breast cancer in Tunisian patients. AB - Previous studies have shown the expression WISP3 and RhoC in cell lines of inflammatory breast cancer (IBC). The aim in the current study was to compare the expression of both genes, in biopsy samples collected from Tunisian patients with localized or metastatic breast cancer and patients with IBC. We investigated 127 patients enrolled in Salah Azaiez Institute in Tunis. Using the RT-PCR, we showed the phenotype (WISP3-, RhoC+) is significantly associated with IBC tumors, while the (WISP3+, RhoC-)phenotype is mostly associated to non-IBC tumors. The frequencies of these tumor phenotypes are significantly different between these tumor groups (p = 10(- 7); relative risk or RR = 3.25; confidential interval or CI 95% = 1.90-5.53). Immunohistochemical test revealing the presence of WISP3 and RhoC proteins correlates with the expression in the biopsy of their encoding genes as detected by RT-PCR. In conclusion, it appears that WISP3 and RhoC genes expression status defines a molecular signature of IBC. PMID- 20014944 TI - Sorafenib therapy for metastatic renal carcinoma in patients with low cardiac ejection fraction: report of two cases and literature review. AB - Targeted therapies used in the treatment of metastatic renal cell carcinoma (RCC) are known to have the potential for cardiotoxicity and should be used with caution in patients with cardiac comorbidities. A retrospective review identified two RCC cases treated with sorafenib in the context of preexisting cardiomyopathy. Sorafenib therapy resulted in disease stabilization of progressing RCC for both cases, without worsening of cardiac ejection fraction. Further evaluation of the cardiac safety of sorafenib in patients with cardiomyopathy is warranted. PMID- 20014945 TI - Correlation between argyrophilic nucleolar organizer region staining and brain tumor classification and grading. AB - Distinctions between benign and malignant tumors are less evident in the central nervous system than in other tissues. Since the level of cell proliferation is an important feature in tumor grading, we applied AgNOR in 50 cases of brain tumors with different grades and histological origins in order to check its efficiency in discriminating between benign and malignant cases. We found significant differences between the means of grade I (1.76) and grade IV (2.46) tumors. No significant differences were observed considering the same grading with distinct histological types or age of patients, reinforcing the efficiency of AgNOR. PMID- 20014946 TI - IGF-1 partially restores chemotherapy-induced reductions in neural cell proliferation in adult C57BL/6 mice. AB - Chemotherapeutic agents produce persistent difficulties in memory through an unknown mechanism. We tested the hypothesis that chemotherapeutic agents readily able to cross the blood-brain barrier (cyclophosphamide and fluorouracil), as opposed to those not known to readily cross the barrier (paclitaxel and doxorubicin), reduce neural cell proliferation following chemotherapy. We found that 5-bromo-2-deoxyuridine labeling following chemotherapy given to C57BL/6 mice revealed a similar reduction in neural cell proliferation in the dentate gyrus for all four agents. Insulin-like growth factor 1, a molecule implicated in promoting neurogenesis, counteracted the effects of high doses of chemotherapy on neural cell proliferation. PMID- 20014947 TI - Expression and mutation analysis of her2 in head and neck squamous cell carcinoma. AB - We analyzed mutation and expression status of human epidermal growth factor receptor 2 (Her2) in head and neck squamous cell carcinoma (HNSCC) using single strand conformation polymorphism (SSCP) mutation analysis and immunohistochemistry (IHC). Mutations were absent in all 85 cases. Out of 57 cases available for IHC, Her2 protein expression was negative (0) in 40 tumors (70%). Seventeen tumors (29.8%) expressed Her2, among these 13 tumors (22.8%) showed a weak (+1) expression and 4 (7%) showed a moderate expression (+2), none showed a strong (+3) expression. There was not a significant association between expression and any of the patients' clinical variables or prognosis. Our results suggest that Her2 may not be useful as a molecular target in HNSCC. PMID- 20014948 TI - Influenza circulation and the burden of invasive pneumococcal pneumonia during a non-pandemic period in the United States. AB - BACKGROUND: Animal models and data from influenza pandemics suggest that influenza infection predisposes individuals to pneumococcal pneumonia. Influenza may contribute to high winter rates of pneumococcal pneumonia during non-pandemic periods, but the magnitude of this effect is unknown. With use of United States surveillance data during 1995-2006, we estimated the association between influenza circulation and invasive pneumococcal pneumonia rates. METHODS: Weekly invasive pneumococcal pneumonia incidence, defined by isolation of pneumococci from normally sterile sites in persons with clinical or radiographic pneumonia, was estimated from active population-based surveillance in 3 regions of the United States. We used influenza virus data collected by World Health Organization collaborating laboratories in the same 3 regions in seasonally adjusted negative binomial regression models to estimate the influenza-associated fraction of pneumococcal pneumonia. RESULTS: During approximately 185 million person-years of surveillance, we observed 21,239 episodes of invasive pneumococcal pneumonia; 485,691 specimens were tested for influenza. Influenza circulation was associated with 11%-14% of pneumococcal pneumonia during periods of influenza circulation and 5%-6% overall. In 2 of 3 regions, the association was strongest when influenza circulation data were lagged by 1 week. CONCLUSIONS: During recent seasonal influenza epidemics in the United States, a modest but potentially preventable fraction of invasive pneumococcal pneumonia was associated with influenza circulation. PMID- 20014949 TI - Health-related quality of life and survival among HIV-infected patients receiving highly active antiretroviral therapy: a study of patients in the AIDS Therapy Evaluation in the Netherlands (ATHENA) Cohort. AB - BACKGROUND: Previous studies have shown that health-related quality of life (HRQL) predicts survival in patients infected with human immunodeficiency virus (HIV). However, these studies predated the highly active antiretroviral therapy (HAART) era, included only a few patients receiving HAART, or had a limited duration of follow-up. This study investigates whether HRQL predicts survival among HIV-infected patients receiving HAART. METHODS: HIV-infected patients participating in the focus group of the AIDS Therapy Evaluation in the Netherlands (ATHENA) study and starting or already receiving HAART completed the Medical Outcomes Study HIV Health Survey at study entry (1 May 1998 through 31 December 2000). The physical health summary (PHS) and mental health summary (MHS) scores were calculated. All-cause mortality was established at 31 March 2008. Kaplan-Meier analysis and Cox regression models were performed to predict survival. RESULTS: The median follow-up was 8.4 years. Sixty-six patients (11.8%) died during follow-up. We found a significant relation between quartiles of PHS and survival (P < .001, log-rank test). Of patients with a PHS, 26 (20%) died in quartile 1 (indicating worst HRQL), 17 (13%) died in quartile 2, 10 (8%) died in quartile 3, and 5 (4%) died in quartile 4 (indicating best HRQL) (P< .001). The prediction of PHS on survival was independent of other (clinical) parameters (P< .001). No relation was found between MHS and survival (P= .13). CONCLUSION: Patient-reported HRQL predicted survival among HIV-infected patients receiving HAART. This information could be highly useful for physicians in determining the prognosis of their patients. PMID- 20014950 TI - Etiology of community-acquired pneumonia: increased microbiological yield with new diagnostic methods. AB - BACKGROUND: The microbial etiology of community-acquired pneumonia (CAP) is still not well characterized. During the past few years, polymerase chain reaction (PCR)-based methods have been developed for many pathogens causing respiratory tract infections. The aim of this study was to determine the etiology of CAP among adults-especially the occurrence of mixed infections among patients with CAP-by implementing a new diagnostic PCR platform combined with conventional methods. METHODS: Adults admitted to Karolinska University Hospital were studied prospectively during a 12-month period. Microbiological testing methods included culture from blood, sputum, and nasopharyngeal secretion samples; sputum samples analyzed by real-time quantitative PCR for Streptococcus pneumoniae, Haemophilus influenzae, and Moraxella catarrhalis; nasopharyngeal specimens analyzed by use of PCR; serological testing for Mycoplasma pneumoniae, Chlamydophila pneumoniae, and viruses common in the respiratory tract; and urine antigen assays for detection of pneumococcal and Legionella pneumophila antigens. RESULTS: A microbial etiology could be identified for 67% of the patients (n = 124). For patients with complete sampling, a microbiological agent was identified for 89% of the cases. The most frequently detected pathogens were S. pneumoniae (70 patients [38%]) and respiratory virus (53 patients [29%]). Two or more pathogens were present in 43 (35%) of 124 cases with a determined etiology. CONCLUSIONS: By supplementing traditional diagnostic methods with new PCR-based methods, a high microbial yield was achieved. This was especially evident for patients with complete sampling. Mixed infections were frequent (most commonly S. pneumoniae together with a respiratory virus). PMID- 20014951 TI - Global trends in typhoid and paratyphoid Fever. AB - Typhoid and paratyphoid fever continue to be important causes of illness and death, particularly among children and adolescents in south-central and Southeast Asia, where enteric fever is associated with poor sanitation and unsafe food and water. High-quality incidence data from Asia are underpinning efforts to expand access to typhoid vaccines. Efforts are underway to develop vaccines that are immunogenic in infants after a single dose and that can be produced locally in countries of endemicity. The growing importance of Salmonella enterica serotype Paratyphi A in Asia is concerning. Antimicrobial resistance has sequentially emerged to traditional first-line drugs, fluoroquinolones, and third-generation cephalosporins, posing patient treatment challenges. Azithromycin has proven to be an effective alternative for treatment of uncomplicated typhoid fever. The availability of full genome sequences for S. enterica serotype Typhi and S. enterica serotype Paratyphi A confirms their place as monomorphic, human-adapted pathogens vulnerable to control measures if international efforts can be redoubled. PMID- 20014952 TI - Pharmacokinetics and safety of extemporaneously compounded valacyclovir oral suspension in pediatric patients from 1 month through 11 years of age. AB - BACKGROUND: Valacyclovir provides enhanced acyclovir bioavailability in adults, but limited data are available in children. METHODS: Children 1 month through 5 years of age with or at risk for herpesvirus infection received a single 25 mg/kg dose of extemporaneously compounded valacyclovir oral suspension (n = 57), whereas children 1 through 11 years of age received 10 mg/kg valacyclovir oral suspension twice daily for 3-5 days (herpes simplex virus infection) (n = 28) or 20 mg/kg 3 times daily for 5 days (varicella-zoster virus infection) (n = 27). Blood samples for pharmacokinetic analysis were collected during the 6 h after the first dose. Safety was monitored throughout the studies. RESULTS: Dose proportionality in the maximum observed concentration (C(max)) of acyclovir and the area under the concentration-time curve from time zero extrapolated to infinity (AUC(0-infinity)) existed across the 10 to 20 mg/kg valacyclovir dose range. For children 2 through 5 years of age, an increase in dose from 20 to 25 mg/kg resulted in near doubling of the C(max) and AUC(0-infinity). Among infants 1 through 2 months of age receiving 25 mg/kg, the mean AUC(0-infinity) and C(max) were higher ( approximately 60% and 30%, respectively) than those among older infants and children receiving the same dose. Valacyclovir oral suspension was well tolerated. No clinically significant trends were noted in clinical chemical, hematologic, or urinalysis values from screening to follow-up. CONCLUSIONS: Among children 3 months through 11 years of age, the 20 mg/kg dose of this formulation of valacyclovir oral suspension produces favorable acyclovir blood concentrations and is well tolerated. A dosing recommendation cannot be made for infants <3 months of age because of decreased clearance in this age group. Trial registration. ClinicalTrials.gov identifier: NCT00297206 . PMID- 20014953 TI - Is diet quality an overlooked mechanism for Bergmann's rule? AB - Bergmann's rule (body size increases with latitude) has long interested biologists; however, its mechanism remains unclear. An overlooked mechanism (latitudinal variation in plant quality) might help explain Bergmann's rule. We studied three herbivores. In the field, the planthopper Prokelisia and the sea hare Aplysia, but not the long-horned grasshopper Orchelimum, were larger at high latitudes, following Bergmann's rule. In the laboratory, all three species grew larger or faster on high-latitude plants. High-latitude diets increased Prokelisia size and Aplysia growth rates by 8% and 72%, respectively, enough to explain the increase in field body size toward high latitudes. Therefore, latitudinal variation in herbivore body size could be influenced by latitudinal variation in plant quality, which may directly or indirectly also affect body size in detritivores, parasitoids, and predators. Studies of Bergmann's rule should consider the influence of biotic factors on body size in addition to abiotic factors such as temperature and precipitation. PMID- 20014954 TI - All-cause mortality and competing risks of fatal and nonfatal vascular events in the Italian longitudinal study on aging: impact of lipoprotein(a). AB - Among possible determinants of vascular events, the role of high lipoprotein(a) (Lp[a]) serum levels represents a still uncertain independent risk factor in elderly populations. Moreover, the cumulative incidence of nonfatal vascular events due to high Lp(a) serum levels is conditioned by the competing risk of death from any causes that are a function of age. After a 6.3-year median follow up, we tested the competing risks of all-cause mortality, cumulative fatal nonfatal stroke events, cumulative fatal-nonfatal coronary artery disease (CAD) events, and nonfatal stroke or CAD events due to high Lp(a) serum levels in a population-based, prospective study conducted in one of the eight centers of the Italian Longitudinal Study on Aging (ILSA), Casamassima, Bari, Italy. Of 704 elderly individuals (65-84 years), 372 (169 women and 203 men) agreed to participate in the study. As compared with those in the lowest Lp(a) tertile serum levels, subjects in the highest tertile (>20 mg/dL) had a higher partially adjusted risk of nonfatal CAD (hazard ratio, 4.19; 95% confidence interval [CI], 1.36-12.94) and nonfatal stroke (hazard ratio, 3.38; 95% CI, 1.00-11.56). Compared with those in the lowest tertile, subjects in the highest tertile had a higher fully adjusted risk of nonfatal CAD (hazard ratio, 3.41; 95% CI, 1.08 10.78). Finally, overall no statistically significant association was found between Lp(a) and the risk of all-cause mortality, cumulative fatal-nonfatal stroke, and cumulative fatal-nonfatal CAD events. In our population, Lp(a) was not a significant independent predictor of stroke and death from all causes, but it was an independent predictor of nonfatal CAD. Finally competing risk, conditioning the timing and occurrence of vascular events in our study population, could be a correct approach for evaluating the role of Lp(a) lipoprotein in vascular disease among elderly people. PMID- 20014955 TI - Gene-gene interactions of the brain-derived neurotrophic-factor and neurotrophic tyrosine kinase receptor 2 genes in geriatric depression. AB - Brain-derived neurotrophic-factor (BDNF) and its receptor neurotrophic tyrosine kinase receptor 2 (NTRK2) have been implicated in both major depression and cognitive function. This study examines the main effects of single loci and multilocus interactions to test the hypothesis that the BDNF and NTRK2 genes may contribute to the etiology of geriatric depression independently and/or through complex interactions. We genotyped the BDNF gene Val66Met (rs6265) polymorphism and four single-nucleotide polymorphisms (SNPs) (including rs1187323, rs1187329, rs1778929, and rs1545285) in the NTRK2 gene in 155 elderly inpatients diagnosed with major depression and 195 age- and sex-similar control subjects. All patients were assessed with the Hamilton Rating Scale for Depression (HAM-D) for depression severity and the Mini-Mental Status Examination (MMSE) for cognitive function after admission. The genotype distributions of all five SNPs tested were significantly different between depressed patients and control subjects. BDNF rs6265, NTRK2 rs1187323, and NTRK2 rs1778929 (p = 0.0031, 0.002, and 0.0014, respectively) also displayed statistically significant differences in the genotypic tests after Bonferroni correction (p < 0.05/5 = 0.01). In addition, the 2-marker haplotype derived from the rs1187323 and rs1187329 polymorphisms demonstrated a significant difference between geriatric depression and control groups according to haplotype distribution (global p = 0.003). Furthermore, BDNF and NTRK2 interactions were found in the significant 2-locus, 3-locus, 4-locus, and 5-locus gene-gene interaction models (p = 0.014, <0.001, 0.007, and 0.032, respectively) using a generalized multifactor dimensionality reduction (GMDR) method. Analyses using logistic regression models confirmed the gene-gene interactions. The results suggest that the BDNF and NTRK2 genes may contribute to the risk of geriatric depression independently and in an interactive manner. PMID- 20014956 TI - Risperidone-related reversal of primary enuresis: an unusual case report. AB - Occasional reports have documented treatment-emergent enuresis with medication regimens that include risperidone. In contrast to these reports, we present a 12 year-old girl with mild mental retardation and primary nocturnal enuresis who received risperidone (0.5 mg/day) for the isolated symptom of inappropriate smiling. Surprisingly, in addition to reduction in inappropriate smiling, risperidone also substantially decreased the frequency of enuresis. These benefits with risperidone were confirmed in an on-off-on treatment sequence. To our knowledge, this is the first case in literature of primary enuresis responding to low-dose risperidone. Low-dose risperidone may merit study in children with enuresis. Clinical implications and possible mechanisms are discussed. PMID- 20014957 TI - Laboratory reporting of hemostasis assays: the final post-analytical opportunity to reduce errors of clinical diagnosis in hemostasis? AB - The advent of modern instrumentation, with associated improvements in test performance and reliability, together with appropriate internal quality control (IQC) and external quality assurance (EQA) measures, has led to substantial reductions in analytical errors within hemostasis laboratories. Unfortunately, the reporting of incorrect or inappropriate test results still occurs, perhaps even as frequently as in the past. Many of these cases arise due to a variety of events largely outside the control of the laboratories performing the tests. These events are primarily preanalytical, related to sample collection and processing, but can also include post-analytical events related to the reporting and interpretation of test results. The current report provides an overview of these events, as well as guidance for prevention or minimization. In particular, we propose several strategies for the post-analytical reporting of hemostasis assays, and how this may provide the final opportunity to prevent serious clinical errors in diagnosis. This report should be of interest to both the laboratory scientists working in hemostasis and clinicians that request and attempt to interpret the test results. Laboratory scientists are ultimately responsible for these test results, and there is a duty to provide both accurate and precise results to enable clinicians to manage patients appropriately and to avoid the need to recollect and retest. Also, clinicians will not be in a position to best diagnose and manage their patient unless they gain an appreciation of these issues. PMID- 20014958 TI - Stability of serum thyroid hormones following 8-11 years of cold storage. AB - BACKGROUND: Ethnic differences necessitate a need for local reference intervals (RI), but establishing these can be challenging in some cultures that are reluctant to donate blood. Frozen sera are an alternative, but results can be questionable. Between 1998 and 2001, we collected blood samples from 343 healthy pregnant Chinese women (5-41 weeks' gestation), and in 2001 published gestational RI for thyrotropin thyroid stimulating hormone (TSH), free thyroxine (FT4) and free triiodothyronine (FT3) using the ACS180, now obsolete. As a pilot to re establishing RI, TSH, FT4 and FT3 were re-assayed in archived specimens using contemporary assays. METHODS: Thirty archived specimens (-80 degrees C) with ACS180 TSH concentrations ranging from 0.25 to 3.7 mIU/L were measured using the Roche E170, Advia Centaur and Architect i2000 TSH, FT4 and FT3 assays, along with 10 newer contemporary samples. Results were compared to the original, and examined in context of past and present methodological performances in an external quality assurance (EQA) program. RESULTS: All contemporary assays detected significantly lower TSH and increased FT4 and FT3 concentrations in the stored samples. CONCLUSIONS: With reference to methodological performances in EQA, the results obtained with contemporary assays suggest analyte deterioration in specimens and thus their unsuitability for re-establishing RI. PMID- 20014963 TI - Editorial. PMID- 20014960 TI - Pathogenic mechanisms in systemic lupus erythematosus. AB - Systemic lupus erythematosus (SLE) is a chronic inflammatory disease characterized by the dysfunction of T cells, B cells, and dendritic cells and by the production of antinuclear autoantibodies. This editorial provides a synopsis of newly discovered genetic factors and signaling pathways in lupus pathogenesis that are documented in 11 state-of-the-art reviews and original articles. Mitochondrial hyperpolarization underlies mitochondrial dysfunction, depletion of ATP, oxidative stress, abnormal activation, and death signal processing in lupus T cells. The mammalian target of rapamycin, which is a sensor of the mitochondrial transmembrane potential, has been successfully targeted for treatment of SLE with rapamycin or sirolimus in both patients and animal models. Inhibition of oxidative stress, nitric oxide production, expression of endogenous retroviral and repetitive elements such as HRES-1, the long interspersed nuclear elements 1, Trex1, interferon alpha (IFN-alpha), toll-like receptors 7 and 9 (TLR 7/9), high-mobility group B1 protein, extracellular signal-regulated kinase, DNA methyl transferase 1, histone deacetylase, spleen tyrosine kinase, proteasome function, lysosome function, endosome recycling, actin cytoskeleton formation, the nuclear factor kappa B pathway, and activation of cytotoxic T cells showed efficacy in animal models of lupus. Although B cell depletion and blockade of anti-DNA antibodies and T-B cell interaction have shown success in animal models, human studies are currently ongoing to establish the value of several target molecules for treatment of patients with lupus. Ongoing oxidative stress and inflammation lead to accelerated atherosclerosis that emerged as a significant cause of mortality in SLE. PMID- 20014959 TI - Regulation of autoreactive B cell responses to endogenous TLR ligands. AB - Immune complexes containing DNA and RNA are responsible for disease manifestations found in patients with systemic lupus erythematosus (SLE). B cells contribute to SLE pathology through BCR recognition of endogenous DNA- and RNA- associated autoantigens and delivery of these self-constituents to endosomal TLR9 and TLR7, respectively. B cell activation by these pathways leads to production of class-switched DNA- and RNA-reactive autoantibodies, contributing to an inflammatory amplification loop characteristic of disease. Intriguingly, self-DNA and RNA are typically non-stimulatory for TLR9/7 due to the absence of stimulatory sequences or the presence of molecular modifications. Recent evidence from our laboratory and others suggests that B cell activation by BCR/TLR pathways is tightly regulated by surface-expressed receptors on B cells, and the outcome of activation depends on the balance of stimulatory and inhibitory signals. Either IFNalpha engagement of the type I IFN receptor or loss of IgG ligation of the inhibitory FcgammaRIIB receptor promotes B cell activation by weakly stimulatory DNA and RNA TLR ligands. In this context, autoreactive B cells can respond robustly to common autoantigens. These findings have important implications for the role of B cells in vivo in the pathology of SLE. PMID- 20014964 TI - Digital infrared and ultraviolet photography using advanced camera services modified equipment. AB - As part of a series of investigations into the utility of digital SLR cameras for IR and UV photography the authors tested a range of Nikon cameras modified by Advanced Camera Services, together with the company's adapted electronic flash units and camera bracket. PMID- 20014965 TI - From pamphlets to podcasts: health information at Great Ormond Street Hospital for Children NHS trust. AB - This paper outlines the development of information for children and families at Great Ormond Street Hospital for Children NHS Trust. Starting in 1995, guidelines for producing written information have been developed and a regular audit programme carried out, resulting in information of consistently high quality being produced. The 'rules' set out for the provision of written information have now been developed to cover multimedia information, so that technology can be harnessed to develop podcasts and other non-written information materials. Technical processes are explained both for production and delivery. Future plans for multimedia information are also outlined. PMID- 20014967 TI - Mohs micrographic surgery. PMID- 20014968 TI - Videoconferencing: live procedures during conferences--practical problems and solutions. PMID- 20014974 TI - Acute epilepsy exacerbations in patients switched between A-rated anti-epileptic drugs. AB - BACKGROUND: Concerns have been raised over the use of different manufacturers' versions of A-rated antiepileptic drug (AED) formulations in epilepsy patients. OBJECTIVE: To estimate the association between acute epilepsy exacerbations and switching between different A-rated AEDs. METHODS: A nested case-control study was conducted using pharmacy and medical claims data from January 1, 2005 through December 31, 2007. 18-65-year-olds who had an epilepsy diagnosis and received AED therapy during 2005 were eligible for study. Cases were defined as individuals with a documented exacerbation of epilepsy in the form of a 2006 or 2007 inpatient or emergency room claim for epilepsy. Controls were from the same population and matched on baseline epilepsy diagnosis and follow-up time since January 1, 2006. The exposure was a switch between A-rated AEDs in the 90 days prior to the matching date. Conditional logistic regression was used to estimate the odds of an epilepsy exacerbation after a switch controlling for important covariates. RESULTS: A total of 34 216 individuals were eligible for study, of whom 2949 cases were matched to 8847 controls. The unadjusted odds ratio (OR) between a switch and an epilepsy exacerbation was 1.51 (95% CI: 1.29-1.76). After adjusting for potential confounders, the odds ratio was 1.08 (95% CI: 0.91-1.29). Treatment with three or more AEDs or a change in outpatient diagnosis from baseline resulted in ORs of 2.96 (95% CI: 2.48-3.49) and 2.53 (95% CI: 2.28 2.82), respectively. CONCLUSIONS: After addressing potential confounders, no evidence that A-rated switching was associated with increased acute exacerbations of epilepsy was found. Study limitations include potentially incomplete identification of seizures, no information on indication for medication use, and limited information on duration and severity of disease. This study provides additional insight into the relationship between A-rated AED switching and acute exacerbations of epilepsy. PMID- 20014975 TI - RAGE-independent autoreactive B cell activation in response to chromatin and HMGB1/DNA immune complexes. AB - Increasing evidence suggests that the excessive accumulation of apoptotic or necrotic cellular debris may contribute to the pathology of systemic autoimmune disease. HMGB1 is a nuclear DNA-associated protein, which can be released from dying cells thereby triggering inflammatory processes. We have previously shown that IgG2a-reactive B cell receptor (BCR) transgenic AM14 B cells proliferate in response to endogenous chromatin immune complexes (ICs), in the form of the anti nucleosome antibody PL2-3 and cell debris, in a TLR9-dependent manner, and that these ICs contain HMGB1. Activation of AM14 B cells by these chromatin ICs was inhibited by a soluble form of the HMGB1 receptor, RAGE-Fc, suggesting HMGB1-RAGE interaction was important for this response. To further explore the role of HMGB1 in autoreactive B cell activation, we assessed the capacity of purified calf thymus HMGB1 to bind dsDNA fragments and found that HMGB1 bound both CG-rich and CG-poor DNA. However, HMGB1-DNA complexes could not activate AM14 B cells unless HMGB1 was bound by IgG2a and thereby able to engage the BCR. To ascertain the role of RAGE in autoreactive B cell responses to chromatin ICs, we intercrossed AM14 and RAGE-deficient mice. We found that spontaneous and defined DNA ICs activated RAGE+ and RAGE(- ) AM14 B cells to a comparable extent. These results suggest that HMGB1 promotes B cell responses to endogenous TLR9 ligands through a RAGE-independent mechanism. PMID- 20014976 TI - Atherosclerosis in systemic lupus erythematosus. AB - Systemic lupus erythematosus (SLE) is an autoimmune rheumatic disease that has a late mortality phase owing mainly to cardiovascular manifestations. Atherosclerosis itself is characterized by inflammatory components, fulfilling the criteria of Witebsky and Rose for an autoimmune disease. SLE patients have increased risk for cardiovascular events, and these are the result of both atherosclerosis and thromboembolic events. Risk factors for atherosclerosis in SLE include "traditional" risk factors (mainly the Framingham risk factors), as well as disease-related factors including disease duration, steroid therapy, and renal disease, and inflammatory mechanisms that specifically contribute to enhanced atherosclerosis in SLE. These include specific antibodies to beta2GPI; anticardiolipin antibodies; anti-oxidized low-density lipoprotein; and antibodies to heat shock proteins, complement activation, impaired ability to activate TGF beta1, and elevated levels of CRP. These findings stress the importance of surveillance and preventive strategies to control atherosclerosis in SLE. PMID- 20014978 TI - Determination of reactive oxygen species associated with the degeneration of dopaminergic neurons during dopamine metabolism. AB - Oxidative stress is believed to be an important mechanism underlying dopamine induced neuronal damage. This study provides X-band electron spin resonance (ESR) spectroscopic evidence for reactive oxygen species (ROS) generation during dopamine metabolism. The authors induced excess dopamine metabolism in the mouse striatum by bathing it in tyramine-containing perfusate using microdialysis. The addition of tyramine to the perfusate raised the levels of extracellular dopamine and hydrogen peroxide significantly. The ESR signal from hydroxy-TEMPO decayed during tyramine perfusion and treatment with a monoamine-oxidase inhibitor or radical scavenger suppressed the signal decay. Decreases in the number of tyrosine hydroxylase-immunopositive fibres and in dopamine concentration after tyramine perfusion were observed. Moreover, the tyramine-perfused mice showed a marked methamphetamine-induced rotational response. Notably, these effects of tyramine were suppressed by the simultaneous perfusion of hydroxy-TEMPO. These findings indicate that the ROS generation, which was monitored by hydroxy-TEMPO, caused oxidative damage to the dopaminergic neurons. PMID- 20014979 TI - Impact of loss of mobility on instrumental activities of daily living and socioeconomic status in patients with MS. AB - OBJECTIVE: To assess the effects of mobility loss on instrumental activities of daily living (IADL) and socioeconomic status in multiple sclerosis (MS) patients. METHODS: Participants were active registrants in the North American Research Committee on Multiple Sclerosis registry completing the Fall 2006 (IADL analysis, n = 10,396) or Spring 2007 (socioeconomic analysis, n = 8180) surveys. Cross sectional correlations and linear and logistic regression were performed using sociodemographic factors, mobility scales, and Patient Determined Disease Steps as independent variables and IADLs as the response. RESULTS: Mobility loss was significantly correlated with decreased IADL scores (r = -0.74; p < 0.0001); this correlation remained significant after adjustment for covariates. Mobility loss also negatively correlated with employment (r = -0.48 for women; r = -0.50 for men, both p < 0.0001) and annual income (r = -0.29; p < 0.0001). These correlations were all significant even with mild mobility loss. The relationships derived from the regression models suggest that the effect of mobility on employment is greater than the effect of demographic variables, and a small but direct effect on annual income that is independent of effects mediated through employment. The self-reported diagnosis of MS for study inclusion and use of single-item ordinal scales for mobility and disability can potentially be criticized as study limitations, although the diagnosis and the scales were previously validated. CONCLUSION: Mobility loss independently correlated with IADL, and associated with reduced socioeconomic status in people with MS. These correlations were significant with mild mobility loss, supporting early treatment. PMID- 20014977 TI - Autoantibody-dependent and autoantibody-independent roles for B cells in systemic lupus erythematosus: past, present, and future. AB - It has long been known that B cells produce autoantibodies and, thereby, contribute to the pathogenesis of many autoimmune diseases. Systemic lupus erythematosus (SLE), a prototypic systemic autoimmune disorder, is characterized by high-circulating autoantibody titers and immune-complex deposition that can trigger inflammatory damage in multiple organs/organ systems. Although the interest in B cells in SLE has historically focused on their autoantibody production, we now appreciate that B cells have multiple autoantibody-independent roles in SLE as well. B cells can efficiently present antigen and activate T cells, they can augment T cell activation through co-stimulatory interactions, and they can produce numerous cytokines which affect inflammation, lymphogenesis, and immune regulation. Not surprisingly, B cells have become attractive therapeutic targets in SLE. With these points in mind, this review will focus on the autoantibody-dependent and autoantibody-independent roles for B cells in SLE and on therapeutic approaches that target B cells. PMID- 20014980 TI - Iron supplementation in the non-dialysis chronic kidney disease (ND-CKD) patient: oral or intravenous? AB - BACKGROUND: The management of iron-deficiency anaemia in patients with non dialysis chronic kidney disease (ND-CKD) remains controversial, particularly regarding the use of oral versus intravenous iron supplementation. METHODS: A PubMed search from 1970 to February 2009 was conducted to identify relevant research articles. FINDINGS: Iron supplementation is advisable for all iron deficient CKD patients receiving erythropoiesis stimulating agents (ESAs), and intravenous iron may be preferable to oral iron. However, there is also a growing body of data indicating that iron supplementation may avoid or delay the need for ESA therapy in some ND-CKD patients. In each of four randomised trials that included ND-CKD patients without ESA, the haemoglobin response was greater with i.v. versus oral iron. Moreover, some ND-CKD patients who remain anaemic on oral iron may subsequently respond to i.v. iron. Newer preparations (ferric carboxymaltose and ferumoxytol) permit rapid, high-dose administration. In a randomised study, a single 15-minute injection of ferric carboxymaltose, with up to two additional doses as required, resulted in 53.2% of ND-CKD patients achieving > or =1 g/dL increase in haemoglobin by day 56 without ESA, compared to 29.9% of patients given oral iron supplements. Two large, randomised, ongoing trials will address the important question of whether i.v. or oral iron supplementation affects the progression of renal dysfunction. While i.v. iron is more costly than oral iron, the cost differential over time may be lower than widely believed, and i.v. therapy avoids the poor absorption, gastrointestinal intolerance and non-compliance associated with oral preparations. In terms of safety, true anaphylaxis does not occur with modern preparations such as iron sucrose and iron gluconate. The novel preparations ferric carboxymaltose and ferumoxytol do not require a test dose and appear to offer a good safety profile, but long-term safety monitoring is mandatory. CONCLUSIONS: Intravenous iron offers an effective, feasible route towards reducing the heavy burden of iron deficiency anaemia in the non-dialysis CKD patient, even in the absence of ESA therapy. PMID- 20014981 TI - Treatment retention with risperidone long-acting injection: 24-month results from the Electronic Schizophrenia Treatment Adherence Registry (e-STAR) in six countries. AB - OBJECTIVE: To assess treatment retention on risperidone long-acting injection (RLAI) and outcomes in schizophrenia patients for whom 24 months of follow-up data in the electronic Schizophrenia Treatment Adherence Registry (e-STAR) were available. RESEARCH DESIGN AND METHODS: e-STAR is an ongoing, international, multicenter, prospective, observational registry assessing use of antipsychotics in patients with schizophrenia or schizoaffective disorder in a normal clinical practice setting. Parameters were assessed prior to and post-initiation of RLAI. Data presented are from six European countries that enrolled patients in e-STAR after they initiated treatment with RLAI. MAIN OUTCOME MEASURES: Clinical and demographic information were collected at baseline and treatment-related data, including RLAI discontinuation, psychiatric hospitalization and medication utilization, were collected prospectively every 3 months. Data collection continued for 24 months, even for patients who discontinued RLAI therapy. Hospitalization and medication utilization were also collected retrospectively by chart review for the 12-month period prior to RLAI initiation. RESULTS: A total of 1659 patients (mean age, 39.2; 18.3% inpatients) completed the study. Twenty four months after initiating therapy (initial RLAI dose = 33.6 mg) 85% of patients (n = 1410) remained on RLAI (completers) while 15% discontinued therapy. The main reasons for discontinuation were insufficient response (28.5%), patient/family choice (26.1%), adverse events (9.6%) and unacceptable tolerability (6.0%). At baseline, compared to completers, discontinuers were younger (37.4 vs. 39.6 years, p = 0.01), had schizophrenia for a shorter time (10.2 vs. 11.9 years, p = 0.02), had lower Global Assessment of Functioning (GAF) scores (43.5 vs. 48.0, p = 0.0001), higher utilization of benzodiazepines (56.5 vs. 43.3%) and more initiated therapy as inpatients (30 vs. 16%). With RLAI therapy GAF scores improved significantly (p < 0.001) for both groups but the 24 month value for discontinuers was lower than that of completers (55.4 vs. 67.2). Compared to the pre-RLAI initiation period, at 12 months post-initiation completers had greater reductions than discontinuers in the percent of patients hospitalized (66.2% reduction vs. 29.2%) and in the length (68% reduction vs. 0%) and number (80.0 vs. 14.3%) of hospital stays, differences that remained at 24 months. The most common adverse events while patients were taking RLAI were nervous system disorders (6.8%), psychiatric disorders (5.6%), weight increase (3.2%), reproductive system and breast disorders (2.5%) and gastrointestinal disorders (2.1%). CONCLUSIONS: These observational data confirm that RLAI is an effective treatment in schizophrenia and high levels of adherence to therapy offers an opportunity for effective long-term disease management and significant sustained decreases in hospitalization. PMID- 20014982 TI - The obesity paradox and outcome in heart failure: is excess bodyweight truly protective? PMID- 20014984 TI - Recent advances in cardiology. AB - During 2009, multiple major cardiology trials have been presented or published. In this paper, we summarize and place in clinical context the new trial findings regarding anticoagulation (dabigatran), antiplatelet therapy (ticagrelor, clopidogrel, prasugrel and aspirin), percutaneous coronary management (thrombectomy, multivessel/left main disease and biodegradable polymers), medical therapy for coronary disease (ivabradine and rosuvastatin) and management of heart failure (beta-blocker strategy, atrial fibrillation and resynchronization therapy). PMID- 20014985 TI - Evaluation of imatinib mesylate in the treatment of pulmonary arterial hypertension. AB - Imatinib mesylate is a small molecule inhibitor that selectively inhibits the PDGF receptor kinase as well the cKIT and Abl kinases, among other targets. Various studies have implicated the PDGF pathway in the pathogenesis of pulmonary arterial hypertension (PAH). Inhibition with imatinib mesylate has shown efficacy in human case reports and experimental models of PAH. Results from a Phase II trial of imatinib mesylate in PAH did not meet the primary end point but showed improvement in several secondary end points and in a subgroup analysis. As suggested by this study as well as a few case reports, imatinib may be effective in a subset of patients with more severe disease. However, this remains to be further validated through a Phase III study, which is already underway. In conclusion, it appears that imatinib mesylate may hold promise as an adjunct drug in PAH therapy, especially since it is directed at a pathway not previously targeted. PMID- 20014986 TI - Echocardiography in 2009: the future of clinical diagnosis. AB - Over the past 10 years we have observed a tremendous expansion on cardiovascular imaging with breathtaking images observing all aspects of the cardiac anatomy and functions. So much so that each imaging modality has grown to the point that one single individual cannot possess all the required expertise for its optimal use and the need for subspecialization in the various imaging modalities is crucial. Echocardiography has surprised many for its ability to adapt in the ever increasing demand and the discovery of the many new modalities. These include 3D and deformation imaging, as well as contrast utilization during the studies, which will continue to put echocardiography in a league of its own in terms of patients management. Some basic descriptions of the various modalities are summarized and their contribution to a number of clinical scenarios, today and in the future, are explored. PMID- 20014987 TI - Pulmonary vein isolation for the treatment of atrial fibrillation: past, present and future. AB - Atrial fibrillation is the most common sustained cardiac arrhythmia and is associated with impaired quality of life and an increased risk of stroke and mortality. Antiarrhythmic drugs, currently the first-line therapy for patients without permanent atrial fibrillation, fail to prevent recurrences in over 50% and are associated with significant adverse effects. In the majority of cases, atrial fibrillation arises from the repetitive firing of myocytes in muscle sleeves that extend from the left atrium into the pulmonary veins. Pulmonary vein isolation has emerged as an effective treatment in selected patients. In this paper, the past and present roles of pulmonary vein isolation are described and the future developments of this technique are explored. PMID- 20014988 TI - Adjuvant therapy for atrial fibrillation. AB - Atrial fibrillation (AF) is the most common heart rhythm disorder, with increasing prevalence in the aging US population and affecting more than 2.3 million people. Current approaches for managing AF are rate- or rhythm-control strategies, both using anti-thrombotic therapy to prevent thromboembolism. While great advances have been made in understanding the pathophysiology of AF, few new strategies have shown promise in prevention or treatment of AF. Recent data suggest that non-antiarrhythmic medication may be useful in modifying the substrate that allows AF precipitation and perpetuation. This article reviews the data on the role of these agents in the prevention and management of AF as an adjunct to standard therapy. PMID- 20014989 TI - Transcatheter aortic valve implantation: what has been done and what is going to be done. AB - Transcatheter valve implantation is developing into an effective and reproducible therapy for aortic valve stenosis. The origin of this technique was pursued in 1992 when Andersen demonstrated the feasibility of percutaneous implantation of catheter-based valve prosthesis. Since then a lot of technical and device advances have been made and to date, transcatheter aortic valve implantation has became a concrete alternative to surgical replacement. This paper aims to go over all the current devices, from the most widely used to the newest technology, focusing on device description, procedural issues, potential complications and clinical studies currently available in literature. PMID- 20014990 TI - Current controversies in heart failure with a preserved ejection fraction. AB - Heart failure with a preserved ejection fraction is a fascinating and multifaceted condition that has provoked enormous debate and a wealth of mechanistic studies. Controversies exist with regard to its nomenclature. If its nomenclature is questioned, one can be certain the pathogenesis is ill understood. If the pathogenesis is disputed, the diagnosis becomes difficult and inconsistent. These diagnostic challenges result in inappropriate recruitment to clinical trials. Therefore, the trials may be underpowered and difficult to interpret. This paper examines contemporary theories of heart failure with a preserved ejection fraction, clarifies the controversies and attempts to resolve the divergences of opinion. PMID- 20014991 TI - Imaging modalities in cardiac electrophysiology. AB - Cardiac imaging, both noninvasive and invasive, has become a crucial part of evaluating patients during the electrophysiology procedure experience. These anatomical data allow electrophysiologists to not only assess who is an appropriate candidate for each procedure, but also to determine the rate of success from these procedures. This article incorporates a review of the various cardiac imaging techniques available today, with a focus on atrial arrhythmias, ventricular arrhythmias and device therapy. PMID- 20014992 TI - Choosing an angiotensin-receptor blocker: blood pressure lowering, cardiovascular protection or both? AB - Angiotensin-receptor blockers (ARBs) offer superior tolerability to angiotensin converting enzyme inhibitors, and are increasingly used in patient management. ARBs vary in their pharmacological profiles, which results in efficacy differences. Therefore, deciding between ARBs should be evidence-based and related to the specific requirements of the individual patient. For patients with hypertension but at low additional risk, an ARB that provides sustained, powerful 24-h reductions in blood pressure is suitable. For patients at very high additional risk (with heart failure), an ARB with demonstrated efficacy in this patient population is the preferred option. For patients at increased risk, telmisartan should be the ARB of choice based on the results from the Ongoing Telmisartan Alone and in Combination with Ramipril Global End Point Trial (ONTARGET), which demonstrated for the first time that an ARB has equivalent protection to the reference angiotensin-converting enzyme inhibitor in a broad cross-section of at-risk patients but a better side-effect profile. PMID- 20014994 TI - Cost-effectiveness analysis of the use of a high-intensity statin compared to a low-intensity statin in the management of patients with familial hypercholesterolaemia. AB - OBJECTIVES: To estimate, using probabilistic decision-analytic modelling techniques, the cost effectiveness of treating familial hypercholesterolaemia (FH) patients with high-intensity statins compared to treatment with low intensity statins. For the purpose of this economic analysis, and based on their known differences, statins were categorised as high intensity if they produce greater LDL-cholesterol reductions than simvastatin 40 mg (e.g., simvastatin 80 mg and appropriate doses of atorvastatin and rosuvastatin or combination of statins + ezetimibe). METHODS: A lifetime Markov model was developed to estimate the incremental cost per quality adjusted life year (QALY) of treating a hypothetical cohort of 1000 FH patients aged between 20 and 70 years. Baseline coronary heart disease risks reported in the NICE TA 94 on statins, and age adjusted risk of cardiovascular disease reported in the FH population, were used to populate the model. A meta-analysis estimate of the reduction in cardiovascular events from using high-intensity compared with low-intensity statins was obtained from published trials. Results were interpreted using a cost effectiveness threshold of pound20 000/QALY. RESULTS: Fewer cardiovascular events and deaths were predicted to occur in the group treated with higher-intensity statins, and the incremental cost-effectiveness ratio (ICER) was estimated at pound11 103/QALY. The ICER remained below the pound20 000 threshold for 20-39 year-olds and 40-59-year-olds, but rose above this threshold in individuals aged over 60 years. One-way sensitivity analysis showed that results were most sensitive to variation in treatment effect on mortality and the cost of high intensity statins. CONCLUSIONS: Modelling demonstrates that high-intensity statins are cost-effective for the treatment of younger FH patients. If, as is likely, the relative price of high-intensity statins fall in the future as they come off patent, then their cost effectiveness will improve further. PMID- 20014995 TI - Mixed treatment comparison and meta-regression of the efficacy and safety of prostaglandin analogues and comparators for primary open-angle glaucoma and ocular hypertension. AB - OBJECTIVES: Primary open-angle glaucoma (POAG) is a chronic condition characterised by optic neuropathy and vision loss. Elevated intraocular pressure (IOP) can damage the optic nerve and is a risk factor for glaucoma, thus treatment usually comprises topical hypotensives. This analysis aims to address methodological issues associated with the synthesis of glaucoma clinical trial data, given variations in study methodology and IOP measurement. METHODS: Meta regression was used to estimate how IOP varies over time for patients receiving treatment. Relative treatment effects were assessed using a random-effects mixed treatment comparison (MTC) in order to preserve randomisation and avoid selection bias. To produce clinically meaningful outputs, these analyses were combined to obtain the mean on-treatment IOP and the proportion of patients achieving different IOP targets at different time points. A further MTC estimated the probability of hyperaemia events. RESULTS: The analysis showed that after 3 months' treatment, between 58 and 83% of patients will have a > or =20% reduction in IOP and 70-93% of patients will have an absolute IOP <20 mmHg. Latanoprost and bimatoprost were found to produce significantly lower on-treatment IOP compared with timolol (p < 0.05); the difference between latanoprost and bimatoprost was not significant. Travoprost produced a lower mean IOP compared with timolol (not significant). Latanoprost-timolol was found to produce significantly lower IOP than latanoprost alone or beta-blockers. The probability of hyperaemia-type events varied between treatments from 14.8 to 63.03%. Latanoprost had significantly lower odds of hyperaemia than travoprost, bimatoprost, travoprost timolol, or bimatoprost-timolol. CONCLUSION: This analysis suggests that latanoprost and bimatoprost produce a statistically significant reduction in IOP compared with timolol, but are associated with a higher risk of hyperaemia. Out of all the prostaglandins, latanoprost may achieve a good balance between tolerability and IOP efficacy. As with all forms of meta-analysis, the results are based on the assumption that the studies and intervention groupings are sufficiently similar to be compared. PMID- 20014996 TI - In vitro reconstruction of an autologous, watertight, and resistant vesical equivalent. AB - PURPOSE: Currently, bladder repair is performed using gastrointestinal segments; however, this technique has a high morbidity rate, and new alternatives are thus needed. The lack of native or synthetic tissue with similar properties of the bladder led us to develop autologous vesical substitutes entirely made by tissue engineering and without exogenous matrices. Watertight function and mechanical resistance are fundamental for the model. The aim of this study was to determine the structural and functional characteristics of our vesical equivalent (VE). MATERIALS AND METHODS: Porcine VEs are produced in 55 days. The cellular types that make up the vesical wall are extracted and purified simultaneously from a small porcine bladder biopsy. Dermal fibroblasts are extracted and cultured in vitro to form cellular sheets. Endothelial cells were seeded on the fibroblast sheets before their superimposition. Urothelial cells are then seeded onto this cellular construction. VEs are characterized by histology, immunostaining, electron microscopy, and cell viability. Mechanical properties of the reconstructed substitutes are evaluated by uniaxial tensile tests, and tissue absorption is verified with (14)C-urea, which quantifies the degree of impermeability. RESULTS: This process allowed us to obtain a highly structured tissue with a total fusion of the fibroblast layers. As expected, histological observations showed a pseudostratification of the urothelium developing on an organized self-secreted extracellular matrix. Positive markers for cytokeratin 8/18 in immunostaining confirmed the presence of a urinary epithelium. Electron microscopy confirmed the normal aspect of urothelial cells. Our VE's permeability to (14)C-urea was significantly similar to porcine bladder, and characterization of the mechanical properties indicated that our tissue could be suitable for grafting since its ultimate tensile strength compares favorably with a native porcine bladder. CONCLUSION: The construction of a VE using this method seems very promising in meeting the needs in the urological field. Our substitute has proven its efficiency as a barrier to urea and has a sufficient mechanical resistance to support suturing. Additionally, this model is completely autologous, and its possible endothelialization could promote the early vascularization process after grafting and thus significantly reducing inflammation and possible rejection. PMID- 20014998 TI - DJ-1 loss by glutaredoxin but not glutathione depletion triggers Daxx translocation and cell death. AB - Environmental and genetic causes are implicated in the etiopathogenesis of Parkinson's disease (PD), a neurodegenerative movement disorder. DJ-1, a putative gene recessively linked to early onset PD, functions as an antioxidant, transcriptional co-activator, and molecular chaperone. We examined DJ-1 status following global perturbation of protein thiol homeostasis by depleting cellular antioxidant glutathione or downregulating glutaredoxin 1, a thiol disulfide oxidoreductase, wherein both paradigms generate oxidative stress. While these perturbations did not affect expression of DJ-1 mRNA, downregulation of glutaredoxin 1 but not glutathione depletion caused loss of DJ-1 protein, translocation of Daxx (a death-associated protein) from nucleus, and cell death. Overexpression of wild-type DJ-1, but not the cysteine mutants, prevented Daxx translocation and cytotoxicity. Protease inhibitors prevented constitutive DJ-1 loss. Residual DJ-1 was present in reduced state, indicating that DJ-1 when oxidized was degraded through proteolysis. Thus, loss of DJ-1 occurring through its oxidative modification and subsequent proteolysis mediated through dysregulation of thiol disulfide oxidoreductase may contribute to pathogenesis of sporadic PD, thus providing a link between environmental challenges and constitutive levels of this vital protein. PMID- 20014999 TI - Targeting interleukin-21 in immune-mediated pathologies. AB - Interleukin (IL)-21, a cytokine mostly produced by activated CD4+ T cells, has been reported to play an important role in the tissue-damaging immune response in various organs. This pathogenic effect is strictly linked to the ability of IL-21 to control the functional activities of multiple immune and non-immune cells. For instance, IL-21 regulates the differentiation and function of effector CD4+ T helper cells; controls activation, proliferation, and survival of B cells and enhances the cytotoxic activity of CD8+ T cells and NK cells. IL-21 also inhibits the differentiation of inducible regulatory T cells (Tregs) and makes effector CD4+ T cells resistant to the Tregs-mediated immunesuppression. Additionally, IL 21 stimulates epithelial cells and fibroblasts to make chemokines and extracellular matrix proteases, respectively. Consistently, studies from various laboratories have documented the beneficial effect of IL-21 neutralization on the progression of inflammatory diseases in mice. Here we review the present knowledge on the expression and role of IL-21 in immune-mediated pathologies. PMID- 20015002 TI - The type II transmembrane serine protease, matriptase-2: Possible links to cancer? AB - Matriptase-2 is a newly identified member of the Type II Transmembrane Serine Protease (TTSP) family. The expression profile of many members of this family of proteases is frequently altered in cancerous cells and tissues and a number of TTSPs have been linked to cancer progression and development. Matriptase-2 is structurally similar to matriptase-1, a TTSP which has gained recent interest due to its potential to enhance the aggressive nature of cancer cells and its links with a variety of human cancers. Recently, matriptase-2 has been functionally linked to the regulation of iron metabolism; however, there is also evidence to suggest that, as with other members of the TTSPs, matriptase-2 may have a role in cancer development and progression. This article reviews the current literature on matriptase-2, together with its potential roles in physiological and disease states particularly focusing on cancer. PMID- 20015003 TI - Application of biological study for met expression to cancer therapy. AB - Metastasization is an undesirable process in cancer development and may represent the most critical factor in deciding patient prognosis. Organ specificity of the metastasis process suggests the importance of the paracrine factors: one of the most potent paracrine regulators of tumor cell migration is hepatocyte growth factor/scatter factor (HGF/SF). Because the liver-specific growth factor is HGF, its receptor c-Met expression might play a critical role in metastasization to the liver. Activation of HGF/c-Met signaling has been shown to promote cancer cell invasiveness and trigger metastasis though direct involvement of the angiogenic pathway. Given the importance of aberrant HGF/c-Met signaling, several different therapeutic strategies aimed at inhibiting the pathway have been developed and are currently being evaluated in clinical trials. Among these agents, NK4 and AM102 were introduced as HGF inhibitors, and PHA-665752 and Su11274 as c-Met inhibitors and are under study in clinical trials. Further, clinical experience-based study to apply the accumulation of biological knowledge concerning HGF/c-Met to the surgical field is presented. PMID- 20015004 TI - Hepatocyte growth factor activation inhibitors - therapeutic potential in cancer. AB - Hepatocyte growth factor (HGF) plays a plethora of roles in the progression of many invasive and metastatic cancers. The interaction between tumour cells and their surrounding stromal environment remains a crucial factor governing tumour invasion and metastasis. HGF is primarily synthesised by stromal fibroblasts as an inactive precursor known as pro-HGF. A number of proteases have demonstrated the ability to convert pro-HGF into the biologically active form of HGF, although the two main factors responsible are HGF activator (HGFA) and matriptase. The HGF activation inhibitors (HAI-1 and HAI-2) are two novel Kunitz-type serine protease inhibitors that regulate HGFA and matriptase activity to govern the influence of HGF within the body. Deregulation of HAI expression can lead to shift in the HGF activation/inhibition balance ratio in favour of enhanced HGF production. Therefore, these HGF activation inhibitors may have a direct bearing on cancer invasion and metastasis. This review examines the accumulating evidence on the emerging role and therapeutic potential of HAI-1 and HAI-2 in cancer. PMID- 20015005 TI - Anti-cancer approach with NK4: Bivalent action and mechanisms. AB - We identified NK4, the N-terminal and four kringle domains of hepatocyte growth factor (HGF), as a specific inhibitor of HGF. NK4 binds to the Met/HGF receptor, but does not activate the Met receptor, thereby competitively inhibiting the HGF Met pathway. Independent of its inhibition of HGF-Met, NK4 acts as an angiogenesis inhibitor. The angioinhibitory action of NK4 is mediated by perlecan, a multidomain proteoglycan involved in vascular basement membrane assembly. The extracellular binding of NK4 to perlecan inhibits cell-associated assembly of fibronectin, and the impaired fibronectin assembly suppresses integrin-dependent angiogenic responses, i.e., endothelial cell proliferation, migration and tube formation. NK4 or an NK4-like fragment is generated by proteases expressed in inflammatory cells, suggesting regulation of physiological or pathological processes by NK4 or NK4-like fragments. In a variety of cancer models, NK4 exhibited anti-cancer effects due to its bifunctional characteristics, including inhibition of invasion and metastasis, inhibition of angiogenesis-dependent tumor growth, and promotion of survival. Several lines of strategies and different molecules that inhibit the HGF-Met pathway have been developed, including small molecular inhibiters of Met tyrosine kinase. The biological action of NK4 as an angiogenesis inhibitor has definite advantages over other molecules. In addition to the well-acknowledged role of HGF-Met in cancer invasion and metastasis, recent studies indicate that activation of the HGF-Met pathway makes tumor-initiating cells invasive and resistant to chemical and radiation therapy. Treatment with NK4 could offer a new therapeutic option for the inhibition of cancer metastasis and growth, and better outcomes for cancer patients. PMID- 20015007 TI - Discovery of small molecule c-Met inhibitors: Evolution and profiles of clinical candidates. AB - The scatter factor/hepatocyte growth factor (HGF)-c-Met axis is involved in the malignant phenotype of various tumor types via activation of a wide range of autocrine and paracrine processes. Autocrine activation of tumor cell c-Met receptors enhances tumor cell proliferation, angiogenesis, invasion/metastasis and resistance to apoptosis and cytotoxic therapies. In addition, tumor and stroma cell-derived HGF functions as a potent angiogenic factor. Therefore, the HGF-c-Met axis is critically involved in multiple facets of normal cellular growth and homeostasis and activated in a dysregulated manner in a variety of cancers. Consequently, inhibiting the HGF-c-Met axis would be anticipated to have potent anti-tumor effects in many cancers through multiple complimentary mechanisms including increased sensitivity to current cytotoxic chemo-and radiotherapies. The acceptance of c-Met as a tractable target for cancer therapy has fostered intensive drug discovery efforts across the pharmaceutical industry. This research has led to 20 published crystal structures (with and without ligands) that revealed two distinct binding modes for ATP-competitive inhibitors: Type I ligands which assumes a U shape geometry through interactions with both hinge and activation loop residue Y1230, and Type II ligands which adopt a more extended orientation, either binding a conventional DFG-out conformation or protein conformations with varying degrees of 'DFG-out' character. Nearly a dozen small molecule c-Met inhibitors have entered human clinical trials ranging from Type I inhibitors solely selective for c-Met to Type I inhibitors with broader kinase activities to Type II inhibitors with "spectrum selective" kinase activity. The identification, profiles and properties of these clinical candidates are summarized in this review. PMID- 20015006 TI - An orally bioavailable c-Met kinase inhibitor potently inhibits brain tumor malignancy and growth. AB - The receptor tyrosine kinase, c-Met and its ligand hepatocyte growth factor (HGF) are important regulators of malignancy in human cancer including brain tumors. c Met is frequently activated in brain tumors and has emerged as a promising target for molecular therapies. Recently, an orally bioavailable small molecule kinase inhibitor of c-Met (SGX523) was developed by SGX Pharmaceuticals. We tested the effects of this inhibitor on c-Met brain tumor cell activation, c-Met-dependent malignancy, and in vivo glioma xenograft growth. SGX523 potently inhibited c-Met activation and c-Met-dependent signaling at nanomolar concentrations in glioma cells, primary gliomas, glioma stem cells and medulloblastoma cells. SGX523 treatment inhibited c-Met-dependent brain tumor cell proliferation and G1/S cell cycle progression. SGX523 also inhibited brain tumor cell migration and invasion. Furthermore, systemic delivery of SGX523 via oral gavage to mice bearing orthotopic human glioblastoma xenografts led to a significant decrease of in vivo tumor growth. These studies show that c-Met activation and c-Met-dependent brain tumor cell and stem cell malignancy can be inhibited by small molecules. The study also shows for the first time that oral delivery of a small molecule kinase inhibitor of c-Met inhibits intracranial tumor growth. These findings suggest that targeting c-Met with small molecule kinase inhibitors is a promising approach for brain tumor therapy. PMID- 20015008 TI - Hepatocyte growth factor and its receptor signalling complex as targets in cancer therapy. AB - Hepatocyte growth factor (HGF) and it's receptor, cMET, have become the focus on intense scrutiny since its discovery in the late 1980s [1 2] as regards it role in cancer. HGF is now known to be a potent morphogen that can regulate tissue and organ regeneration and modulate cell morphology, it is a motogen that can stimulate cell motility and migration, and is a mitogen able to regulate cell growth and death and as an angiogenic factor [3]. HGF is can induce both angiogenesis and lymphangiogenesis [3a; 4]. That HGF and cMET have such a diversity of functions has led to intense interest in the clinical setting due to their potential in their prognostic aspect and therapeutic implications as imaging tools. This issue will focus on recent work that shows strong indications for the value of HGF and cMET in clinical settings. PMID- 20015009 TI - Silibinin--a promising new treatment for cancer. AB - Silymarin and its major constituent, Silibinin, are extracts from the medicinal plant Silybum marianum (milk thistle) and have traditionally been used for the treatment of liver diseases. Recently, these orally active, flavonoid agents have also been shown to exert significant anti-neoplastic effects in a variety of in vitro and in vivo cancer models, including skin, breast, lung, colon, bladder, prostate and kidney carcinomas. The aim of the present review is to examine the pharmacokinetics, mechanisms, effectiveness and adverse effects of silibinin's anti-cancer actions reported to date in pre-clinical and clinical trials. The review will also discuss the results of current research efforts seeking to determine the extent to which the effectiveness of silibinin as an adjunct cancer treatment is influenced by such factors as histologic subtype, hormonal status, stromal interactions and drug metabolising gene polymorphisms. The results of these studies may help to more precisely target and dose silibinin therapy to optimise clinical outcomes for oncology patients. PMID- 20015010 TI - Gestational trophoblastic neoplasia, an ancient disease: new light and potential therapeutic targets. AB - Gestational trophoblastic neoplasia is a rare malignancy, which can occur after any type of pregnancy. The incidence varies according to the geographical location and ethnic origin. Although most patients with gestational trophoblastic neoplasia are cured by conventional chemotherapy and surgery, some suffer resistant disease and may die. New therapeutic agents are needed to reduce the toxicity associated with conventional chemotherapy and treat those with resistant or refractory disease. Molecular targeted treatment provides an exciting avenue, however, the biology of gestational trophoblastic neoplasia is not well understood. This review briefly summarises the recent advances in understanding the pathogenesis and molecular biology of this group of diseases and sheds light on molecules that could provide potential therapeutic targets. PMID- 20015011 TI - gamma-Secretase as a therapeutic target in Alzheimer's disease. AB - gamma-Secretase is an intramembranous multi-protein complex that cleaves many type-I proteins with critical roles in neuronal function. In Alzheimer's disease (AD) interest in gamma-secretase comes, in part, from the fact that this complex is responsible for the last cleavage step of the amyloid precursor protein (APP) that generates the amyloid-beta peptide (Abeta). Abeta represents the primary component of the amyloid plaque, one of the main pathological hallmarks of AD. Over the last years, considerable efforts have been made to develop drugs to reduce Abeta production with the aim to slow AD progression. Many inhibitors of this protease have been identified, although the clinical use has been limited by concerns about the possible toxicity of these compounds. gamma-secretase inhibitors have been shown to reduce Abeta in vitro and in vivo, but interference with Notch proteolysis causes immunological and gastrointestinal toxicity in animal models. The observation that some nonsteroidal anti-inflammatory drug (NSAID) derivatives are able to specifically lower Abeta42 and the development of inhibitors with Notch-sparing selectivity has revived the interest in gamma secretase as an attractive target for drug intervention in AD. Despite the fact that all clinical trials with NSAIDs or gamma-secretase modulators in AD have failed to show clinical benefit thus far, the main concern is that the Abeta lowering potency of the tested compounds may be too low. Active efforts are being made to develop compounds able to penetrate into the brain to lower Abeta at physiological doses without interfering with the cleavage and function of other critical gamma-secretase substrates. These novel inhibitors and modulators may soon offer hope in the Alzheimer's fight. PMID- 20015012 TI - The epithelial-mesenchymal transition and the estrogen-signaling in ovarian cancer. AB - Epithelial ovarian cancer is the leading cause of death for gynecological cancer in most of the Western world; lethality ensues from the occurrence of occult metastasis within the peritoneal cavity, a process requiring the acquisition of capacity for migration and invasiveness by ovarian tumor cells (metastatic phenotype), and characterized by a complex series of interrelated cellular events. Unlike most carcinomas that dedifferentiate during neoplastic progression with loss of epithelial E-cadherin (epithelial to mesenchymal transition, EMT), ovarian carcinomas undergo transition to a more epithelial phenotype, early in tumor progression, with increased E-cadherin expression. Subsequent reacquisition of mesenchymal features is observed in late-stage tumors, and loss of E-cadherin expression or function is a factor in ovarian cancer progression. Changes in E cadherin expression are indicative of the phenotypic plasticity that occurs in ovarian cancer, with a variety of signal transduction pathways impinging on the regulation of E-cadherin levels or subcellular distribution. Among them, the Snail transcription family, consisting of members SNAIL and SLUG, is thought to be mainly involved in the repression of E-cadherin expression, leading to EMT. E cadherin, SNAIL, and SLUG also represent crucial targets of estrogen signaling. In this review, we discuss recent advances in the understanding of the role of estrogen signaling in the complex network underlying the phenotypic plasticity in ovarian cancer. Insight into the mechanisms involved will allow rational drug designs, aimed at the molecules critical to cellular signaling. PMID- 20015013 TI - Combinatorial design of avian influenza neuraminidase inhibitors containing pyrrolidine core with a reduced susceptibility to viral drug resistance. AB - Using computer-assisted combinatorial chemistry techniques, we have designed a virtual library of anti-influenza agents, analogs of inhibitor A-315675, containing a novel pyrrolidine core, which effectively inhibits both wild type and common oseltamivir-resistant mutant forms of the neuraminidase (NA) subtype N1 of avian influenza virus H5N1. A target-specific Potential of Mean Force (PMF) scoring function parameterized on a training set of 13 known pyrrolidine-based inhibitors of NA and validated on 3 others was used to predict the N1 inhibition constants for the focused library of A-315675 analogs. Nine virtual hits (best pyrrolidine inhibitors designed in the present study) are predicted to exhibit inhibition constants in the low picomolar range, up to 200 fold lower than the parent inhibitor A-315675 while displaying favorable predicted ADME-related properties. Proposed small highly-focused combinatorial subsets composed of R groups most frequently occurring in the 200 most active analogs can be useful as a guide for synthetic and medicinal chemists who are developing a new generation of drugs against the avian influenza virus H5N1 by focusing their attention to this small portion of the chemical space. PMID- 20015014 TI - Fluorescent substrates useful as high-throughput screening tools for ADAM9. AB - Fluorescence resonance energy transfer substrates were designed and tested as substrates for ADAM9. The donor/quencher pair used were 5-carboxy fluorescein (Fam) and 4-(4-dimethyl-aminophenylazo)benzoyl (Dabcyl) since they have been well studied sensitive fluorescent probes. The peptides based on precursor TNF-alpha, Dabcyl-Ser-Pro-Leu-Ala-Gln-Ala-Val-Arg-Ser-Ser-Lys(Fam)- NH2 and Dabcyl-Leu-Ala Gln-Ala-HomoPhe-Arg-Ser-Lys(Fam)- NH2, and C-terminal TGF-alpha, Dabcyl-Glu-His Ala-Asp-Leu-Leu-Ala-Val-Val-Ala-Ala-Lys(Fam)- NH2 cleavage sites were effectively processed by ADAM9 with turnover numbers of 100 +/- 20 x 10(-2) min(-1), 20 +/- 10 x 10(-2) min(-1), and 10 +/- 3 x 10(-2) min(-1). In addition, a peptide based on the 33 kDa cleavage site of the low affinity receptor for IgE, CD23, Dabcyl Leu-Arg-Ala-Glu-Gln-Gln-Arg-Leu-Lys-Ser-Lys(Fam)- NH2 was processed as well but with less efficiency. A more selective substrate for ADAM9 was found based on the betacellulin cleavage site. However, the valine containing precursor TNF-alpha based substrate was used to measure IC50 values of metalloproteinase inhibitors against ADAM9 since it was processed the most efficiently. The tightest binding inhibitor was the Wyeth Aerst compound, TMI-1, with an IC50 of 2.1 +/- 0.3 nM. In addition, GI254023, previously identified as a selective inhibitor of ADAM10, also inhibited ADAM9 with an IC50 of 280 +/- 110 nM. These results demonstrate that sensitive substrates for ADAM9 can be developed that are useful in high throughput screening assays for ADAM9. PMID- 20015015 TI - High throughput calorimetry for evaluating enzymatic reactions generating phosphate. AB - A calorimetric assay is described for the high-throughput screening of enzymes that produce inorganic phosphate. In the current example, cellobiose phosphorylase (EC 2.4.1.20) is tested for its ability to synthesise rare disaccharides. The generated phosphate is measured in a high-throughput calorimeter by coupling the reaction to pyruvate oxidase and catalase. This procedure allows for the simultaneous analysis of 48 reactions in microtiter plate format and has been validated by comparison with a colorimetric phosphate assay. The proposed assay has a coefficient of variation of 3.14% and is useful for screening enzyme libraries for enhanced activity and substrate libraries for enzyme promiscuity. PMID- 20015016 TI - High throughput screening of normal and neoplastic tissue samples. AB - The capacity to rapidly and efficiently elucidate a reliable set of disease specific biomarkers is paramount to enable a future of personalized medicine. High throughput screening methods applied to human clinical samples for the discovery of diagnostic, prognostic, and therapeutic targets address this need. Although the ability to analyze either thousands of markers from one sample or one marker from thousands of samples is the current state of high throughput screening, it would be ideal to have the ability to analyze thousands of markers from thousands of samples to expedite the early discovery phase of biomarkers and their validation. This review summarizes the current state of high throughput screening of tissue specimens and discusses its applications. In addition, the rationale, difficulties, strategies, and development of new technologies to address the need for improved high throughput capabilities are discussed. PMID- 20015017 TI - High-throughput binding analysis determines the binding specificity of ASF/SF2 on alternatively spliced human pre-mRNAs. AB - High-throughput immunoprecipitation studies of transcription factors and splicing factors have revolutionized the fields of transcription and splicing. Recent location studies on Nova1/2 and Fox2 have identified a set of cellular targets of these splicing factors. One problem with identifying binding sites for splicing factors arises from the transient role of RNA in gene expression. The primary role of most splicing factors is to bind pre-mRNA co-transcriptionally and participate in the extremely rapid process of splice site selection and catalysis. Pre-mRNA is a labile species with a steady state level that is three orders of magnitude less abundant than mRNA. As many splicing factors also bind mRNA to some degree, these substrates tend to dominate the output of location studies. Here we present an in-vitro method for screening RNA protein interactions that circumvents these problems. We screen approximately 4000 alternatively spliced exons and the entire Hepatitis C genome for binding of ASF/SF2, the only splicing factor demonstrated to function as an oncogene. From the pre-mRNA sequences returned in this screen we discovered physiologically relevant ASF recognition element motifs. ASF binds two motifs: a C-rich and a purine rich motif. Comparisons with similar data derived from the hnRNP protein PTB reveals little overlap between strong PTB and ASF/SF2 sites. We illustrate how this method could be employed to screen disease alleles with the set of small molecules that have been shown to alter splicing in search for therapies for splicing diseases. PMID- 20015018 TI - Miniaturisation of a high throughput screening assay comparing air displacement and capillary-based nanolitre transfer technologies. AB - The miniaturisation of high-throughput screening (HTS) assays has been a widely debated and researched strategy with the aims of reducing screening costs and increasing speed while not compromising data quality. In this context, liquid handling technologies continue to be improved. A new and promising development is the emergence of nanolitre dispensers, which are capable of direct compound transfer to assay microplates. In this study, we investigated the miniaturisation of a HTS kinase assay and compared the real life performance of current state-of the-art air displacement transfer technology (MiniTrak V System) and a capillary based nanolitre dispenser (CyBi-HummingWell). The robustness and effectiveness of the miniaturised assay formats were compared by testing staurosporine to generate dose-response curves and 340 previously identified active compounds. Using the MiniTrak device, assay miniaturisation was achieved from 18 microL to 6 microL in 384-well and 1536-well plate formats. Utilising the nanolitre dispenser, miniaturisation was performed down to 5 microL in 1536-well plates. The Z' factors obtained for each assay format were consistently above 0.5. The data presented here describe the reproducibility of the results obtained with the two transfer technologies and highlight possible issues for hit identification. PMID- 20015022 TI - Efficient growth inhibition of human osteosarcoma cells using a peptide derived from the MDM-2-binding site of p53. AB - Protein transduction therapy is a promising alternative to gene therapy, but has not previously been investigated for osteosarcoma. We here demonstrate efficient growth inhibition of human osteosarcoma cells using a p53 peptide. Our result suggests that protein transduction therapy has significant potential as a novel therapeutic approach for osteosarcoma. PMID- 20015023 TI - Novel detection system for plant protein production of pharmaceuticals and impact on conformational diseases. AB - tate-of-the-art biochemistry methods in combination with an automated phenotyping method demonstrate the high potential of transgenic tobacco plants in producing properly-folded therapeutic proteins for the treatment of protein-misfolding diseases (e.g., Alzheimer's disease). This molecular farming approach led to highest protein production of hydroponically-grown tobacco compared to other growth substrates generally used in plant cultivation. PMID- 20015024 TI - Importance of a potential salt bridge and hydrophobic core in the function and oligomerization of a small heat shock protein. AB - SsHSP14.1, a novel sHSP from the hyper-thermophilic archaeon Sulfolobus solfataricus (S. solfataricus), is reported herein to function to protect EcoR I from heat-induced inactivation. A predicted salt bridge and hydrophobic interactions were found to be important for this function. PMID- 20015025 TI - Primary structural documentation of the major urinary protein of the Indian commensal rat (Rattus rattus) using a proteomic platform. AB - PURPOSE: A number of proteome studies have been performed recently to identify pheromone-related protein expression and their molecular function using genetically modified rodents' urine. However, no such studies have used Indian commensal rodents; interestingly, in a previous investigation, we confirmed the presence of volatile molecules in commensal rodents urine and these molecules seem to be actively involved in pheromonal communication. Therefore, the present study aims to identify the major urinary protein [MUP] present in commensal rat urine, which will help us to understand the protein's expression pattern and intrinsic properties among the rodents globally. EXPERIMENTAL DESIGN: Initially, the total urinary proteins were separated by 1-D and 2-D electrophoresis and then subsequently analyzed by Matrix Assisted Laser Desorption Ionization-Time of Flight and Mass Spectrometer (MALDI-TOF/MS). Furthermore, they were then fragmented with the aid of a Tandem Mass Spectrometer (TOF/TOF) and the identified sequences aligned and confirmed using similarity with the deduced primary structures of members of the lipocalin superfamily. RESULTS: The SDS-PAGE protein profiles showed distinct proteins with molecular masses of 15, 22.4, 25, 28, 42, 50, 55, 68, and 91 kDa. Of these proteins, the 22.4 kDa protein was considered to be target candidate. When 2D elctrophoresis was carried out, about approximately 50 spots were detected with different masses and various pI ranges. The 22.4 kDa protein was found to have a pI of about 4.9.This 22.4 kDa protein spot was digested and subjected to mass spectrometry; it was identified as rat MUP. The fragmented peptides from the rat MUP at 935, 1026, 1192, and 1303 m/z were further fragmented with the aid of MS/MS and generated de novo sequence and this confirmed this protein to be the MUP present in the urine of commensal rats. CONCLUSION: The present investigation confirms the presence of MUP with a molecular mass of 22.4 kDa in the urine of commensal rats. This protein may be involved in the binding of volatile molecules and opens up a discussion about how volatile and non-volatile molecules in the commensal rats' urine may contribute chemo-communication. PMID- 20015026 TI - One-step purification of functional light-harvesting 2 complex from Rhodobacter sphaeroides. AB - Most common method for isolation of light-harvesting 2 complex (LH2) is very time consuming. In our present work functional LH2 complex was one-step rapidly purified from Rhodobacter sphaeroides by metal-ion affinity chromatography. The purified LH2 complex exhibited high activity in light absorption and energy transfer. Our present work provides an efficient approach for preparation of functional LH2 complex. PMID- 20015029 TI - Novel anti-inflammatory and neuroprotective agents for Parkinson's disease. AB - Parkinson's disease (PD) is a type of motor system disorder that results from the progressive loss of dopaminergic (DAergic) neurons in the substantia nigra (SN) of the midbrain. It is one of the most common neurodegenerative disorders, with an incidence that is second only to Alzheimer's disease (AD). Although replacement of dopamine can temporarily alleviate the symptoms of PD patients, it can not prevent the progression of the disease. Increasing evidence has suggested that neuroinflammation significantly contributes to the progress of PD. Therefore, anti-inflammatory therapy could represent a promising neuroprotective intervention with the potential to delay or prevent onset of the disease. This review summarizes several novel potential agents/candidates that might open new avenues for the treatment of PD. In addition to possessing demonstrated anti inflammatory activities that operate through different molecular mechanisms, these agents exert neuroprotective effects by enhancing the production of neurotrophic factors or interfering with the apoptosis of neurons. PMID- 20015030 TI - Development of nanoparticles for antimicrobial drug delivery. AB - This review focuses on the development of nanoparticle systems for antimicrobial drug delivery. Numerous antimicrobial drugs have been prescribed to kill or inhibit the growth of microbes such as bacteria, fungi and viruses. Even though the therapeutic efficacy of these drugs has been well established, inefficient delivery could result in inadequate therapeutic index and local and systemic side effects including cutaneous irritation, peeling, scaling and gut flora reduction. Nanostructured biomaterials, nanoparticles in particular, have unique physicochemical properties such as ultra small and controllable size, large surface area to mass ratio, high reactivity, and functionalizable structure. These properties can be applied to facilitate the administration of antimicrobial drugs, thereby overcoming some of the limitations in traditional antimicrobial therapeutics. In recent years, encapsulation of antimicrobial drugs in nanoparticle systems has emerged as an innovative and promising alternative that enhances therapeutic effectiveness and minimizes undesirable side effects of the drugs. Here the current progress and challenges in synthesizing nanoparticle platforms for delivering various antimicrobial drugs are reviewed. We also call attention to the need to unite the shared interest between nanoengineers and microbiologists in developing nanotechnology for the treatment of microbial diseases. PMID- 20015031 TI - Antioxidants in treating pathologies involving oxidative damage: an update on medicinal chemistry and biological activity of stobadine and related pyridoindoles. AB - Pathologies involving oxidative stress are indicative of malfunction of endogenous antioxidant capacity. Numerous efforts were made to design and synthesize biologically active antioxidants and free oxygen radical scavenging substances that could improve the endogenous antioxidant status. The antioxidant and reactive-oxygen-species-scavenging activity of STB was well demonstrated in many in vitro and in vivo studies. These properties of STB seem to be closely related to its beneficial effects in models of oxidative-stress-involving pathologies, including myocardial infarction, stroke, neurodegenerative disorders, hypoxic-ischemic tissue injury, diabetic complications, chronic inflammation, etc. STB has a good affinity to lipids and exerts its protective activity against free-radical-mediated damage by preventing lipid peroxidation. Rather than interacting with the radicals initiating lipid peroxidation, STB was shown to act in its propagation stage via scavenging peroxyl and/or alkoxyl radicals. STB was also found to protect proteins, predominantly by a mechanism involving protection of thiol groups and by preventing oxidation of amino acids. The first findings on antioxidant and pharmacodynamic effects of STB, tested in a variety of biological models, were summarized in 1998. Recently, chemical modification of STB, which we considered as the leading structure, led to the synthesis of pyridoindole derivatives with significantly increased intrinsic antiradical activity and overall antioxidant efficacy compared to the parental molecule. The present paper provides a complete overview of the literature published since 1998 on both STB and STB congeners. Moreover, appropriate structural modifications of STB provided the opportunity to modulate lipophilicity and acidobasic behavior, thus optimizing bioavailability of the novel derivatives and attenuating their unwanted sideeffects, with the result of decreased toxicity. Hence, STB congeners might be prospectively used as medicinal antioxidants, i.e. remedies effective in conditions involving oxidative stress mediated injury. PMID- 20015032 TI - Blocking nuclear import of pre-integration complex: an emerging anti-HIV-1 drug discovery paradigm. AB - Although recent progress in highly active antiretroviral therapy (HAART) has provided an effective way to treat AIDS patients, the emergence of drug-resistant HIV-1 strains and drug toxicity during long-term treatment of HIV-infected patients necessitate the search for new targets that can be used to develop novel antiviral agents. One such target is the nuclear import process of the HIV pre integration complex (PIC). The ability of HIV-1 using host cell nuclear import machinery to translocate the viral PIC into the cell nucleus is the critical determinant in the replication of the virus in non-dividing cells, such as macrophages. Compounds inhibiting HIV-1 nuclear import may be attractive candidates for novel anti-HIV development. In this review, we will describe the mechanisms of HIV-1 PIC translocation into the nucleus and the structure-function of the viral and cellular factors involved in this process, as well as several classes of novel anti-HIV compounds which target the nuclear import of HIV-1 PIC and effectively block viral replication. PMID- 20015033 TI - A new frontier in the treatment of cancer: NK-1 receptor antagonists. AB - The past two decades have witnessed an exponential increase in research into cancer. This effort, however, has not been translated into better perspectives as regards the problem, although several fields of research have certainly been promising (the human genome project, gene therapy, the search for new cytostatic agents and stem cell research). New pathways must be opened up to offer future hope to oncologic patients. Thus, there is a need to explore other research initiatives in cancer ways to improve this chronic global problem. Substance P (SP) has a widespread distribution in both the central and peripheral nervous systems. It is known that after binding to the specific neurokinin-1 (NK-1) receptor, SP regulates biological functions related to cancer, such as tumour cell proliferation, angiogenesis, and migration of the tumour cells for invasion and metastasis. By contrast, it is also known that after binding to NK-1 receptors, the NK-1 receptor antagonists specifically inhibit tumour cell proliferation (tumour cells die by apoptosis), angiogenesis and the migration of the tumour cells. It is also known that NK-1 receptors are overexpressed in tumours. All these observations suggest that the SP/NK-1 receptor system could play an important role in the development of cancer and metastasis; that the NK-1 receptor could be a new promising target in the treatment of cancer, and that NK 1 receptor antagonists could improve cancer treatment. PMID- 20015034 TI - Age-related pharmacokinetic and pharmacodynamic changes and related risk of adverse drug reactions. AB - Aging is known to be associated with an increased prevalence of multiple chronic diseases, which frequently causes the use of complex therapeutic regimens. The aging process is characterized by relevant changes in drug handling, physiological reserve, and pharmacodynamic response. Hepatic drug clearance of several drugs decreases with aging, mainly due to reduced blood flow, and hepatocyte mass. Renal function also declines with aging, mainly due to sclerotic changes in the glomeruli. Furthermore, due to reduced muscle mass, older subjects frequently have depressed glomerular filtration rate despite normal serum creatinine, and such a concealed renal insufficiency may impact significantly the clearance of hydrosoluble drugs. Changes in pharmacodynamics are also well documented in the cardiovascular and nervous system. Age-related changes in pharmacokinetics and pharmacodynamics, together with comorbidity and polypharmacy, make elderly patients at special risk for adverse drug reactions, which in turn are cause of relevant health burden and costs. Selected measures can assist in preventing or detecting timely such adverse events. PMID- 20015035 TI - Medicinal chemistry of drugs used in diabetic cardiomyopathy. AB - Diabetes mellitus is a common disease and contributes to a high degree of morbidity and mortality. Cardiovascular complications, including diabetic cardiomyopathy are major causes of morbidity and mortality in diabetic patients. Diabetic cardiomyopathy is a condition that affects the myocardium, primarily. It is not necessarily associated with ischemic heart disease, high blood pressure, valvular or congenital anomalies. The pathology of diabetic cardiomyopathy includes interstitial fibrosis, apoptosis of cardiomyocytes, abnormal energy utilization, small vessel disease and cardiac neuropathy. These pathologies are induced by hyperglycemia and oxidative stress. Biochemical as well as electrolyte changes, especially reduced calcium availability also occurs in the myocardium of diabetic patients. The abnormal structure and biochemistry of the myocardium result in functional problems such as diastolic and systolic dysfunctions, which may cause symptoms of dyspnea and inability to tolerate exercise. No single specific therapeutic agent can treat diabetic cardiomyopathy because once the disease is overt, the management may require a variety of approaches such as risk factors and lifestyle modification, glucose control (insulin, alpha glucosidase inhibitors, sulfonylureas, biguanides, meglitinides, thiazolidinediones and dipeptidyl peptidase 4 (DPP-4) inhibitors); hormones (IGF-1); ACE inhibitors (captopril, enalapril); angiotensin II receptor antagonists (losartan, olmesartan); beta adrenoreceptor antagonists (acebutolol, carvedilol); peptides (adrenomedullin); endothelin-1 receptor antagonists (bosentan, tezosentan); calcium channel blockers (amlodipine, verapamil); antioxidants (methalothionein, alpha tocopherol, alpha lipoic acid) and antihyperlipidemic drugs (simvastatin, fenofibrate, ezetimibe) to effectively treat patients with diabetic cardiomyopathy. PMID- 20015036 TI - Mechanisms of action and medicinal applications of abscisic Acid. AB - Since its discovery in the early 1960's, abscisic acid (ABA) has received considerable attention as an important phytohormone, and more recently, as a candidate medicinal in humans. In plants it has been shown to regulate important physiological processes such as response to drought stress, and dormancy. The discovery of ABA synthesis in animal cells has generated interest in the possible parallels between its role in plant and animal systems. The importance of this molecule has prompted the development of several methods for the chemical synthesis of ABA, which differ significantly from the biosynthesis of ABA in plants through the mevalonic acid pathway. ABA recognition in plants has been shown to occur at both the intra- and extracellularly but little is known about the perception of ABA by animal cells. A few ABA molecular targets have been identified in vitro (e.g., calcium signaling, G protein-coupled receptors) in both plant and animal systems. A unique finding in mammalian systems, however, is that the peroxisome proliferator-activated receptor, PPAR gamma, is upregulated by ABA in both in vitro and in vivo studies. Comparison of the human PPAR gamma gene network with Arabidopsis ABA-related genes reveal important orthologs between these groups. Also, ABA can ameliorate the symptoms of type II diabetes, targeting PPAR gamma in a similar manner as the thiazolidinediones class of anti diabetic drugs. The use of ABA in the treatment of type II diabetes, offers encouragement for further studies concerning the biomedical applications of ABA. PMID- 20015037 TI - Bone mass gain during puberty and adolescence: deconstructing gender characteristics. AB - Primary prevention of osteoporosis must aim at increasing bone mass acquisition before late adolescence. During pubertal years both genders reach peak bone acquisition, though males develop a greater skeletal mass. This dimorphism is largely regulated by endocrine factors, with critical roles played by gonadal steroids, growth hormone and insulin growth factor-1, amongst the most important. Menstrual history is a surrogate for the adequacy of hormonal functioning, nutrition and physical activity that may be a marker of bone status and development in young women. Adequate levels of adrenal, reproductive and pituitary hormones, growth factors and leptin are needed for the initiation and maintenance of regular menstrual cycles as well as for the achievement of peak bone mass. Adequate regular exercise and body composition are also pivotal elements in maintaining normal mechanical bone stimulus during bone growth. Avoidance of carbonated soft drink consumption, or excessive alcohol and any tobacco should be considered as these may interfere reaching adequate bone mass. PMID- 20015038 TI - Chagas disease: progress and new perspectives. AB - Chagas disease, also known as American trypanosomiasis, is caused by infection with the protozoan parasite Trypanosoma cruzi. The Pan American Health Organization (PAHO) estimates that currently 7.7 million of people have Trypanosoma cruzi infection in the 21 endemic countries from the southern and southwestern United States to central Argentina and Chile. The only approved therapeutics for the treatment of Chagas disease are two nitroheterocyclic compounds as a nitrofuran (nifurtimox; Lampit) and a nitroimidazole (benznidazole; Rochagan). However, the anti-Trypanosoma cruzi activities of these compounds were discovered empirically over three decades ago. The treatment of Chagas disease with nifurtimox or benznidazole is unsatisfactory because of their limited efficacy in the prevalent chronic stage of the disease and their toxic side effects. In this context, this article will review the current knowledge of the different aspects involved in this illness, such as Trypanosoma cruzi transmission, physiology and biochemistry of the etiological agent, epidemiological aspects and current treatments for American trypanosomiasis. An important section of this review will focus on the different strategies in drug discovery for Chagas disease, including methodology, in vitro screening studies against whole parasites, novel rationally developed approaches on the basis of the increasing knowledge of the biochemistry of Trypanosoma cruzi and the recent progress in the understanding and validation of several targets for the therapy of Chagas's disease. A summary of the most relevant drug targets such as sterol biosynthesis pathway, cysteine protease pathway, pyrophosphate metabolism and purine salvage pathway will be reviewed. Moreover, recent studies regarding other strategies currently under development including thiol-dependent redox metabolism, lysophospholipid analogues and DNA binders will also be discussed. PMID- 20015039 TI - The roles of microRNAs in heart diseases: a novel important regulator. AB - MicroRNAs (miRNAs) are non-coding single-stranded RNAs with about 21~23 nucleotides in length, which originate from encoding genes in nucleus. miRNAs play an inhibitory role in gene expression in a post-transcriptional level by partially complementary binding to the 3' unstranlated region (UTR) of target mRNAs. Large bodies of evidence have shown that miRNAs were involved in various diseases, such as cancer, infectious diseases, diabetes etc, and rising as critical modulators of pathological processes. Lately, some highlight articles revealed that the altered expression of miRNAs such as miR-1, miR-133, miR-21, miR-208 etc in hearts also contributed to cardiovascular diseases, such as heart ischemia, cardiac hypertrophy, and arrhythmias. Moreover, miRNAs were also identified to regulate heart development. These exciting findings not only improve our understanding of the molecular mechanisms of heart diseases, but also provide a new class of potential molecular targets. miRNAs, for the development of novel agents to treat heart diseases. Here, we summarized the recent discoveries about the role of miRNAs in cardiac physiological and pathological functions, and then discussed about their therapeutic potentials for heart diseases. PMID- 20015040 TI - 11beta-hydroxysteroid dehydrogenase type 1 inhibitors as promising therapeutic drugs for diabetes: status and development. AB - Glucocorticoids (GC) play a fundamental role in controlling physiologic homeostasis and, when present in excess, can have a detrimental impact on glucose control, blood pressure and lipid levels. The oxidoreductase 11beta hydroxysteroid dehydrogenase type 1 (11beta-HSD1) mainly catalyzes the intracellular regeneration of active GCs (cortisol, corticosterone) from inert inactive 11-keto forms (cortisone) in liver, adipose tissue and brain, amplifying local GC action. Multiple lines of evidence have indicated that 11beta-HSD1 mediated intracellular cortisol production may have a pathogenic role in type 2 diabetes and its co-morbidities. The 11beta-HSD1 becomes a novel target for anti type 2 diabetes drug developments, and inhibition of 11beta-HSD1 offers a potential therapy to attenuate the type 2 diabetes. In the past several years, a lot of 11beta-HSD1 inhibitors have been designed, synthesized, screened and discovered. Lowering intracellular glucocorticoid concentrations through administration of small molecule 11beta-HSD1 selective inhibitors, significantly attenuates the signs and symptoms of disease in preclinical animal models and clinical trials of diabetes and metabolic syndrome. Among published inhibitors, DIO-902 from DiObex Inc. and INCB13739 from Incyte Inc. are now being investigated under Phase 2B clinical trials. However, the selectivity of current selective 11beta-HSD1 inhibitors has been just focused on the difference between 11beta-HSD1 and 11beta-HSD2. They inhibit the bi-directional activities of 11beta HSD1, both 11beta-HSD1 reductase (major) and oxidase (minor). In our lab, we have recently found novel chemicals that not only inhibit 11beta-HSD1 reductase activity but also increase its oxidase activity without inhibition against 11beta HSD2. We propose that this dual modulation on 11beta-HSD1 may provide a better therapeutic strategy for type 2 diabetes. PMID- 20015041 TI - Neurobiological effects of Hyperforin and its potential in Alzheimer's disease therapy. AB - St. John's Wort (SJW) has been used medicinally for over 5,000 years. Relatively recently, one of its phloroglucinol derivatives, hyperforin, has emerged as a compound of interest. Hyperforin first gained attention as the constituent of SJW responsible for its antidepressant effects. Since then, several of its neurobiological effects have been described, including neurotransmitter re-uptake inhibition, the ability to increase intracellular sodium and calcium levels, canonical transient receptor potential 6 (TRPC6) activation, N-methyl-D-aspartic acid (NMDA) receptor antagonism as well as antioxidant and anti-inflammatory properties. Until recently, its pharmacological actions outside of depression had not been investigated. However, hyperforin has been shown to have cognitive enhancing and memory facilitating properties. Importantly, it has been shown to have neuroprotective effects against Alzheimer's disease (AD) neuropathology, including the ability to disassemble amyloid-beta (Abeta) aggregates in vitro, decrease astrogliosis and microglia activation, as well as improve spatial memory in vivo. This review will examine some of the early studies involving hyperforin and its effects in the central nervous system (CNS), with an emphasis on its potential use in AD therapy. With further investigation, hyperforin could emerge to be a likely therapeutical candidate in the treatment of this disease. PMID- 20015042 TI - New investigational drugs for the management of acute heart failure syndromes. AB - Acute heart failure syndromes (AHFS) enclose a broad spectrum of conditions with different clinical presentations, heart failure history, pathophysiology, prognosis and treatment. AHFS represent a major public health problem because of their high prevalence, high rates of mortality and readmissions and significant healthcare costs, and a therapeutic challenge for the clinicians because management strategies vary markedly. Traditionally used drugs for the treatment of AHFS, including diuretics, vasodilators and positive inotropics, improve clinical signs and symptoms as well as hemodynamics, but present important limitations, as they fail to reduce and may even increase in-hospital and postdischarge mortality, especially in patients with coronary artery disease. Thus, we need new pharmacological agents to not only improve signs and symptoms and cardiac performance, but also improve both short- and long-term outcomes (hospitalizations/survival). In the last decade, significant efforts have been made to identify new therapeutic targets involved in the genesis/progression of AHFS and to develop new therapeutic strategies that may safely improve outcomes. As a result, several new families of drugs have been developed and are currently studied in experimental models and in Phase II and III clinical trials, in an attempt to define their efficacy and safety profiles as well as their precise role in the treatment of AHFS patients. This review firstly analyzes the main clinical applications and limitations of conventional drugs, and then focuses on the mechanisms of action and effects of recently approved drugs and of new investigational agents on signs, symptoms, hemodynamics and outcomes in AHFS patients. PMID- 20015043 TI - Serotonin 5-HT3 and 5-HT4 ligands: an update of medicinal chemistry research in the last few years. AB - The biogenic amine serotonin (5-hydroxytryptamine, 5-HT) is one of the most studied neurotransmitters in the central nervous system. It acts through the activation of at least fourteen 5-HT receptor subtypes. Over the last two decades, high attention was devoted to the 5-HT(3) and 5-HT(4) receptors due to their colocalization in the gastrointestinal tract and because their ligands are useful in the treatment of intestinal serotonergic system dysfunctions. The focus of this review is to discuss the literature concerning recent advances on 5 HT(3)R and 5-HT(4)R ligands and their structure-activity relationships from a medicinal chemistry perspective. During the last few years, new and significant progresses have been made in the field of novel potent and selective ligands, mixed ligands, agonists, partial agonists, and antagonists, and a number of patents have been filed. Furthermore several ligands targeting the 5-HT(3)R and 5 HT(4)R have been proposed for novel therapeutic indications such as the treatment of various psychiatric disorders. PMID- 20015044 TI - Comparative characterization of experimental and calculated lipophilicity and anti-tumour activity of isochromanone derivatives. AB - Compound lipophilicity connected to ADME(T)(a) has great importance in drug development and it has to be evaluated by the generally used drug developmental process. In addition to the importance of lipophilicity in ADMET, recently it has been reported that lipophilicity of small molecules correlates with their antiproliferative activity because of certain specific hydrophobic and lipophilic interactions. Due to the complexity of ADME(T) parameters an efficient and fast method is needed to characterize the many promising candidate lead molecules as a preselection in order not to be rejected from the latter phase of drug development. In the present paper we provide an overview of the importance of lipophilicity of drug candidates for biological action and for ADME(T) and describe a novel approach for drug-likeness characterization of a molecular library using correlation study between lipophilicity and biological activity. Lipophilicity and molecular characteristics have been measured, predicted and optimized for a diverse library from which the best members have been selected to describe their biological, chemical and drug-likeness properties. Molecules were selected from the family of alpha,beta-unsaturated ketones and thorough HPLC characterization for lipophilicity and morphological, antiproliferative and flow cytometric studies were carried out on them. Based on the results 17 member isochromanone library including E and Z geometric isomers were selected for further characterization. In this focused library linear correlation has been found between the calculated and measured lipophilicity and significant parabolic correlation was found between the antiproliferative effect and lipophilicity. Using our efficient and fast method, from a diverse library, we identified an outstandingly effective inhibitor of A431 tumour cell growth via a PARP(a) cleavage dependent apoptosis. In summary the optimized HPLC analyses of lipophilicity combined with the cell-culture assay, introduced above, resulted in the determination of an optimal lipophilicity range. This optimized lipophilicity range should be used in designing novel antiproliferative compounds. PMID- 20015045 TI - Cytotoxic properties of clofibrate and other peroxisome proliferators: relevance to cancer progression. AB - The biological activity of peroxisome proliferators (PPs) is mediated by a class of receptors, known as PPARs (PP-Activated Receptor), belonging to the nuclear receptor superfamily. Upon ligand binding, PPARs dimerize with retinoid receptors, translocate to the nucleus, recognize specific PP-responsive elements on DNA and transactivate a number of genes. Several processes are regulated by PPARs, such as mitochondrial and peroxisomal fatty acid uptake and beta oxidation, inflammation, intracellular lipid trafficking, cell proliferation and death. In addition, PPARs have been proposed to act as tumor suppressors or as tumor promoters, depending on the circumstances. In particular, PPs have been extensively studied for their hepatocarcinogenic action in rodents, most often ascribed to their antiapoptotic action. Recent evidence, however, has been provided about the antiproliferative, proapoptotic, and differentiation-promoting activities displayed by PPAR ligands. The present review will focus on the cytotoxic effects exerted by several PPs, among which clofibrate, on different types of tumor cells, with particular reference to the mechanisms of cell death and to their relevance to cancer induction and progression. PMID- 20015047 TI - Specific recognition of DNA by small molecules. AB - DNA exhibits many attractive features in the development of drug design and disease treatment, and there are some successful events for drugs targeting to DNA in clinic use. Nowadays, the specific recognition of DNA by small molecules is the crucial factor of the sustainable development of DNA-target drugs, for the serious side-effects of existing drugs. In this review, we discussed the current efforts of DNA-drug specific interaction that concerned with various DNA recognition models, such as abasic, mismatch or bulge site recognition, specific sequence recognition, and secondary structure recognition. Some compounds are found to show a remarkable potential to be an ideal drug. PMID- 20015046 TI - Computational analysis of Human Immunodeficiency Virus (HIV) Type-1 reverse transcriptase crystallographic models based on significant conserved residues found in Highly Active Antiretroviral Therapy (HAART)-treated patients. AB - Reverse transcription of the viral single-stranded (+) RNA genome into double stranded DNA is an essential step in the human immunodeficiency virus' (HIV) life cycle. Although several viral proteins are involved in the regulation and/or efficiency of reverse transcription, the process of retroviral DNA synthesis is entirely dependent on the enzymatic activities of the retroviral reverse transcriptase enzyme (RT). Due to its crucial role in the HIV life-cycle, RT is a primary target for anti-HIV drug development. Nonetheless, drug resistance is the major problem affecting the clinical efficacy of antiretroviral agents. Incomplete pharmacological pressure represents the logical cause and not the consequence of different mutation pathways in RT associated with approved inhibitors resistance. In this review we have analyzed RT Protein Data Bank (PDB) models using our innovative computational approach "GRID Based Pharmacophore Model" (GBPM). This method was applied to clinically relevant RT conserved residues found in a large cohort of HAART treated patients. The PDB entries have been selected among the unbound and the complexed models with DNA and/or inhibitors. Such an approach has revealed itself useful to highlight the mutation effects in the drug-RT recognition as well as in the heterodimer stabilization of the enzyme. Most of the clinical and biochemical evidences already reported in the literature have been rationalized at molecular level via the GBPM computational approach. A definite future application of this method will be the identification of conserved regions of critical macromolecules, such as the HIV-1 RT, to be targeted for the development of innovative therapeutic agents. PMID- 20015048 TI - Atrial fibrillation--advances in drug therapy. AB - The prevalence of atrial fibrillation (AF) continues to increase worldwide. It is projected that by 2050, between 5 and 15 million Americans will have AF. Such predictions necessitate new advances in drug therapy. AF is associated with changes in ionic currents. Atrial fibrosis, changes in gap junctions and cell membranes act in concert to initiate and maintain AF. Significant advances in understanding the pathophysiology of this condition have led to numerous new drug discoveries in the last decade. We discuss newly patented molecules that act as sodium and potassium channel blockers, gap junction and intracellular calcium modulators, stretch receptor antagonists, ischemic ion current inhibitors and adenosine receptor agonists. PMID- 20015049 TI - New strategies and drugs in the treatment of hypertension: monotherapy or combination? AB - Hypertension is one of the major risk factors associated with cardiovascular diseases. A range of blood pressure-lowering agents is available including diuretics, alpha- and beta-blockers, aldosterone antagonists, calcium-channel blockers (CCB), angiotensin-converting enzyme inhibitors (ACEI), angiotensin II receptor blockers (ARB) and direct renin inhibitors (DRI). Most patients require two or more medications to control their blood pressures within normal ranges. When high blood pressure cannot be controlled by low-dose monotherapy, physicians employ either high-dose monotherapy or combination therapy. High-dose ARB monotherapy is more effective for reducing proteinuria against low-dose ARB monotherapy or CCBs. Combination therapy is recommended for hypertension patients to facilitate prompt maintenance of blood pressure. Single-pill combination therapy simplifies treatment and optimizes long-term compliance. Thiazide diuretics such as hydrochlorothiazide (HCTZ), alone or in combination are still widely used as first-line hypertension treatment. Recent studies have shown that double (CCB+ARBs) or triple (CCB+ARBs+HCTZ) combination therapies have a greater lowering efficacy and are better tolerated. Moreover, the use of DRIs has been patented and proven effective in selected categories of hypertensive patients with or without concomitant target organ damage (TOD). PMID- 20015050 TI - Pericellular activation of hepatocyte growth factor by the transmembrane serine proteases matriptase and hepsin, but not by the membrane-associated protease uPA. AB - HGF (hepatocyte growth factor) is a pleiotropic cytokine homologous to the serine protease zymogen plasminogen that requires canonical proteolytic cleavage to gain functional activity. The activating proteases are key components of its regulation, but controversy surrounds their identity. Using quantitative analysis we found no evidence for activation by uPA (urokinase plasminogen activator), despite reports that this is a principal activator of pro-HGF. This was unaffected by a wide range of experimental conditions, including the use of various molecular forms of both HGF and uPA, and the presence of uPAR (uPA receptor) or heparin. In contrast the catalytic domains of the TTSPs (type-II transmembrane serine proteases) matriptase and hepsin were highly efficient activators (50% activation at 0.1 and 3.4 nM respectively), at least four orders of magnitude more efficient than uPA. PS-SCL (positional-scanning synthetic combinatorial peptide libraries) were used to identify consensus sequences for the TTSPs, which in the case of hepsin corresponded to the pro-HGF activation sequence, demonstrating a high specificity for this reaction. Both TTSPs were also found to be efficient activators at the cell surface. Activation of pro-HGF by PC3 prostate carcinoma cells was abolished by both protease inhibition and matriptase-targeting siRNA (small interfering RNA), and scattering of MDCK (Madin Darby canine kidney) cells in the presence of pro-HGF was abolished by inhibition of matriptase. Hepsin-transfected HEK (human embryonic kidney)-293 cells also activated pro-HGF. These observations demonstrate that, in contrast with the uPA/uPAR system, the TTSPs matriptase and hepsin are direct pericellular activators of pro-HGF, and that together these proteins may form a pathway contributing to their involvement in pathological situations, including cancer. PMID- 20015051 TI - Euglena gracilis ascorbate peroxidase forms an intramolecular dimeric structure: its unique molecular characterization. AB - Euglena gracilis lacks a catalase and contains a single APX (ascorbate peroxidase) and enzymes related to the redox cycle of ascorbate in the cytosol. In the present study, a full-length cDNA clone encoding the Euglena APX was isolated and found to contain an open reading frame encoding a protein of 649 amino acids with a calculated molecular mass of 70.5 kDa. Interestingly, the enzyme consisted of two entirely homologous catalytic domains, designated APX-N and APX-C, and an 102 amino acid extension in the N-terminal region, which had a typical class II signal proposed for plastid targeting in Euglena. A computer assisted analysis indicated a novel protein structure with an intramolecular dimeric structure. The analysis of cell fractionation showed that the APX protein is distributed in the cytosol, but not the plastids, suggesting that Euglena APX becomes mature in the cytosol after processing of the precursor. The kinetics of the recombinant mature FL (full-length)-APX and the APX-N and APX-C domains with ascorbate and H2O2 were almost the same as that of the native enzyme. However, the substrate specificity of the mature FL-APX and the native enzyme was different from that of APX-N and APX-C. The mature FL-APX, but not the truncated forms, could reduce alkyl hydroperoxides, suggesting that the dimeric structure is correlated with substrate recognition. In Euglena cells transfected with double-stranded RNA, the silencing of APX expression resulted in a significant increase in the cellular level of H2O2, indicating the physiological importance of APX to the metabolism of H2O2. PMID- 20015052 TI - Growth and survival signalling in B16F10 melanoma cells in 3D culture. AB - The two-way communication between the ECM (extracellular matrix) and the cytoplasm via the integrins has many functions in cancer cells, including the suppression of apoptosis. As cells in a 3D (three-dimensional) architecture resemble the in vivo situation more closely than do cells in more conventional 2D cultures, we have employed a substratum that prevents cell adhesion and induces cell aggregation to determine why highly metastatic B16F10 melanoma cells resist anoikis. We compared the behaviour of B16F10 cells in 2D [on tPS (tissue culture polystyrene)] and 3D culture {on polyHEMA [poly(2-hydroxyethylmethacrylate)]} configurations. For this, we analysed cell morphology, proliferation, apoptosis and the activation status of several proteins involved in cell proliferation and survival [RhoA, FAK (focal adhesion kinase), Akt, ERK1/2 (extracellular-signal regulated kinase 1/2)]. B16F10 cells in 3D architecture were able to proliferate as cell aggregates for 3 days, after which the number of cells decreased. The normal Swiss 3T3 cells used as an anoikis-sensitive control did not proliferate on the anti-adhesive substratum. Rho A was activated in B16F10 aggregates throughout their time in culture, whereas it was not in Swiss 3T3 aggregates. An absence of apoptotic activity was correlated with the proliferation of B16F10 cells in aggregates: caspase 3 was significantly activated only after 3 days in culture on polyHEMA. FAK and Akt were transiently activated, and their inactivation was correlated with the induction of apoptosis. ERK1/2 were activated throughout the 3D culture. No survival protein was activated in Swiss 3T3 aggregates. Data obtained from cells in 3D culture suggest that B16F10 cells are resistant to anoikis through the activation of the FAK and Akt signalling pathways. PMID- 20015053 TI - Melasma in men: a clinical, aetiological and histological study. AB - BACKGROUND: Melasma, a facial hypermelanosis, is more common in women. In Indians, men seem to be frequently involved. There are hardly any studies delineating the clinical, aetiological and histological features of melasma in men and the present study was taken up to fill this lacuna. MATERIALS AND METHODS: A total of 200 patients with melasma were screened for only men having melasma. Data including duration, illnesses, sunlight exposure, use of cosmetics, oil or medication, familial pigmentation, nutritional, parasitic infestations, infections, hepatic disorders, occupation were taken followed by general physical, cutaneous and Woods light examination. Laboratory investigations including skin biopsy were performed. RESULTS: Of 200 patients screened, 41 (20.5%) were men. Their ages ranged from 19 to 53 years. Twenty-four (58.5%) of the patients were outdoor workers. Twelve (29.3%) originally belonged to hilly regions. Clinical patterns were malar in 61%, centrofacial in 29.3% and mandibular in 9.7%. The aetiological factors identified were: sun-exposure in 20 (48.8%), mustard oil usage in 18 (43.9%), family history in 16 (37%), chronic illnesses in five (12.2%) and phenytoin in three (7.3%); of these sun-exposure and family history were statistically significant when compared with those for women. Laboratory investigations revealed anaemia in five (12.2%), giardiasis in two (4.9%), increased leuteinizing hormone (LH) and low testosterone in four (9.7%) men. Skin biopsies in 20 (48.8%) patients revealed features of epidermal melasma in 10 (50%) and a mixed type in nine (45%) patients. CONCLUSIONS: Melasma is frequently observed in Indian men. The main causative factors among the male patients appeared to be sun-exposure and family history. Melasma in men is definitely less common than in women, but shares the same clinicohistopathological characteristics as in women. PMID- 20015054 TI - Albright hereditary osteodystrophy: report of a particular clinical phenotype caused by a novel GNAS mutation. PMID- 20015055 TI - Quality of life in patients with lepromatous leprosy in China. AB - BACKGROUND: Leprosy has an impact on patients' quality of life (QoL). However, there has been no study specifically on the impact of the severest type of leprosy-lepromatous leprosy on QoL. OBJECTIVE: To describe the use of Dermatology Life Quality Index (DLQI) among patients with lepromatous leprosy in China. METHODS: Sixty-four inpatients with lepromatous leprosy of Shangluo hospital and Hanzhong hospital and sixty-four controls (healthy volunteers or patients with other dermatoses) matched for age and gender were asked to complete DLQI questionnaires from 2 September 2008 to 20 December 2008. Extensive data were collected besides DLQI, including demographic data and disease-related characteristics. Absence or presence of disability among patients with lepromatous leprosy was evaluated at the same time. RESULTS: The overall mean DLQI score for lepromatous leprosy (18.78) was higher than that for control (9.00) (P < 0.001). Patients with lepromatous leprosy scored significantly higher for all items (P < 0.001) except Q4 (clothes choice). Controls scored significantly lower for all domains of DLQI. Scores of LL increased markedly with increasing clinical severity, but were not associated with educational level, gender, age and disease duration. The inter-item correlation averaged 0.240 and Cronbach's alpha was 0.759, indicating high internal consistency. CONCLUSIONS: This is the first exclusive study to attempt to measure the impact of lepromatous leprosy on QoL. Lepromatous leprosy has a severe impact on QoL. PMID- 20015056 TI - Methotrexate/narrowband UVB phototherapy combination vs. narrowband UVB phototherapy in the treatment of chronic plaque-type psoriasis--a randomized single-blinded placebo-controlled study. AB - BACKGROUND: Psoriasis is a chronic, recurring inflammatory disease affecting the skin, joints and nails that has a significant negative impact on the quality of life. Efficacy of combination of methotrexate/narrowband ultraviolet B (NBUVB) phototherapy in the treatment of psoriasis has been rarely assessed. OBJECTIVES: To compare the efficacy of methotrexate/NBUVB phototherapy combination vs. NBUVB phototherapy in the treatment of chronic plaque psoriasis. METHODS: Forty patients with chronic plaque-type psoriasis (body surface area involvement >10%) were randomized to receive either methotrexate/NBUVB phototherapy (group A) or placebo/NBUVB phototherapy (group B). End point of treatment was 75% reduction in Psoriasis Area and Severity Index (PASI) Score or upto 6 months, whichever was earlier. Patients were then followed up for a period of 12 weeks for assessment of relapse. RESULTS: Of 40 patients, 37 completed the treatment period and 29 both the treatment period and follow-up. PASI 75 was achieved in 19/20 patients in group A and 14/20 patients in group B (P < 0.04). The mean number of weeks(P = 0.001), the mean cumulative dose of NBUVB (P = 0.001) and the mean number of phototherapy sessions (P = 0.0001) required to achieve PASI 75 were significantly less in group A compared with group B. There was no significant difference in the number of patients who relapsed during the follow-up period (P = 0.68). CONCLUSION: Combination of methotrexate and NBUVB phototherapy provides more rapid clinical improvement compared with NBUVB monotherapy in the treatment for chronic plaque-type psoriasis. PMID- 20015057 TI - Porphyria cutanea tarda induced by olmesartan. PMID- 20015058 TI - Professional risk for skin cancer development in male mountain guides--a cross sectional study. AB - OBJECTIVE: For reasons of their outdoor work, mountain guides (MG) are heavily exposed to ultraviolet radiation during their work. METHODS: A standardized interview and examination were performed on 283 male MG (median 41 years) from Germany, Switzerland and Austria and 309 age-matched controls. The median occupation time as MG was 17 years; 39.9% were working full-time. RESULTS: The incidence of skin cancer and precancerous lesions was obtained. Precancerous lesions as solar keratosis (SK) were significantly more frequent in MG (25.4% vs. 7.4%). There was no skin cancer [BCC, SSC, melanoma (MEL)] in the control subjects. Basal cell carcinoma (BCC) was diagnosed in 20 MG (7.1%) and SSC in four MG (1.4%). There were 10 highly suspicious melanocytic lesions; one MG had a histologically confirmed malignant MEL. Risk factors for SK in the multivariate analysis included occupation (P < 0.0001), age (P < 0.0001) and skin type (P = 0.0002). Within the MG group, age (P < 0.0001) and hair colour (P = 0.0058) were independent risk factors for SK. Severe lifetime sunburns (P = 0.0007) and skin type (P = 0.041) were the significant risk factors for BCC, within the MG group in addition to the number of guiding days (P = 0.010). The risk factor for skin cancer (BCC, SCC and MEL) was the number of heavy sunburns during lifetime (P = 0.0014). CONCLUSION: The present study demonstrates an association between high occupational ultraviolet-exposure and an increased prevalence of precancerous skin lesions and skin cancer. MG may be considered as an example for other outdoor professionals. Skin cancer of outdoor workers is likely to be an occupational disease. Primary and secondary prevention should be enforced. PMID- 20015059 TI - The prevalence of red-cell antigens and antibodies in Malawi. AB - As there were no reliable data in Malawi for the prevalence of red cell alloantibodies or antigens in the population, a study was conducted to screen 1000 patients for the presence of antibodies and to type them for ABO, RhD, C, c, E, e and K antigens and to test 500 donors for these antigens plus Fy(a), Fy(b), Jk(a), Jk(b), S and s. Red cell antibodies were identified in 11 patients [1.1%]; 2 were anti-D, 2 anti-S, 1 anti-Le(a+b) and 6 anti-M, 4 of which were found in non-transfused males suggesting they might be naturally acquired. The antigen frequencies found were similar to those previously published for Central Africa but 98.2% of donors were found to be Fy(a-b-). All patients tested were K negative and only three donors were found to be K positive, one being a Caucasian. Approximately 3.5% of Malawians are D negative, lower than the usual 8% quoted for Black Africans. These data confirm the assumption that pre transfusion antibody screening is not currently required but that use of the indirect antiglobulin test in the cross-match is necessary. Haemolytic disease of the newborn (HDN) appears to be rare, or under reported, in Malawi, and more work is needed to find the real incidence of this condition. PMID- 20015060 TI - The development of a national surveillance system for monitoring blood use and inventory levels at sentinel hospitals in South Korea. AB - We developed a web-based program for a national surveillance system to determine baseline data regarding the supply and demand of blood products at sentinel hospitals in South Korea. Sentinel hospitals were invited to participate in a 1 month pilot-test. The data for receipts and exports of blood from each hospital information system were converted into comma-separated value files according to a specific conversion rule. The daily data from the sites could be transferred to the web-based program server using a semi-automated submission procedure: pressing a key allowed the program to automatically compute the blood inventory level as well as other indices including the minimal inventory ratio (MIR), ideal inventory ratio (IIR), supply index (SI) and utilisation index (UI). The national surveillance system was referred to as the Korean Blood Inventory Monitoring System (KBIMS) and the web-based program for KBIMS was referred to as the Blood Inventory Monitoring System (BMS). A total of 30 256 red blood cell (RBC) units were submitted as receipt data, however, only 83% of the receipt data were submitted to the BMS server as export data (25 093 RBC units). Median values were 2.67 for MIR, 1.08 for IIR, 1.00 for SI, 0.88 for UI and 5.33 for the ideal inventory day. The BMS program was easy to use and is expected to provide a useful tool for monitoring hospital inventory levels. This information will provide baseline data regarding the supply and demand of blood products in South Korea. PMID- 20015061 TI - The NAC transcription factor RIM1 of rice is a new regulator of jasmonate signaling. AB - Jasmonates (JAs) are lipid-derived regulators that play crucial roles in both host immunity and development. We recently identified the NAC transcription factor RIM1 as a host factor involved in multiplication of rice dwarf virus (RDV). Here, we report that RIM1 functions as a transcriptional regulator of JA signaling and is degraded in response to JA treatment via a 26S proteasome dependent pathway. Plants carrying rim1 mutations show a phenotype of root growth inhibition. The expression profiles of the mutants were significantly correlated with those of JA-treated wild-type plants without accumulation of endogenous JA, indicating that RIM1 functions as a component of JA signaling. The expression of genes encoding JA biosynthetic enzymes (lipoxygenase (LOX), allene oxide synthase 2 (AOS2) and OPDA reductase 7 (OPR7)) was up-regulated in the rim1 mutants under normal conditions, and a rapid and massive accumulation of endogenous JA was detected in the mutants after wounding. These results suggest that RIM1 may represent a new molecular link in jasmonate signaling, and may thereby provide new insights into the well-established coronatine-insensitive 1 (COI1)-Jasmonate ZIM-domain (JAZ) JA signaling pathway. PMID- 20015062 TI - The MtSNF4b subunit of the sucrose non-fermenting-related kinase complex connects after-ripening and constitutive defense responses in seeds of Medicago truncatula. AB - Dormant seeds are capable of remaining alive in the hydrated state for extended periods of time without losing vigor, until environmental cues or after-ripening result in the release of dormancy. Here, we investigated the possible role of the regulatory subunit of the sucrose non-fermenting-related kinase complex, MtSNF4b, in dormancy of Medicago truncatula seeds. Expression of MtSNF4b and its involvement in a high-molecular-weight complex are found in dormant seeds, whereas imbibition of fully after-ripened, non-dormant seeds leads to dissociation of the complex. MtSNF4b is capable of complementing the yeast Delta snf4 mutant and of interacting with the MtSnRK1 alpha-subunit in a double hybrid system. Transcriptome analyses on freshly harvested and after-ripened RNAi Mtsnf4b and wild-type embryos implicate MtSNF4b in the defense response in hydrated dormant embryonic tissues, affecting the expression of genes encoding enzymes of flavonoid and phenylpropanoid metabolism, WRKY transcription factors and pathogenesis-related proteins. Silencing MtSNF4b also increased the speed of after-ripening during dry storage, an effect that appears to be related to a change in base water potential. No significant difference in ABA content or sensitivity was detected between mutant and wild-type seeds. Pharmacological studies using hexoses and sugar analogs revealed that mannose restored germination behavior and expression of the genes PAL, CHR and IFR in RNAi Mtsnf4b seeds towards that of the wild-type, suggesting that MtSNF4b might act upstream of sugar-sensing pathways. Overall, the results suggest that MtSNF4b participates in regulation of a constitutively activated defense response in hydrated, dormant seeds. PMID- 20015063 TI - Xylem ionic relations and salinity tolerance in barley. AB - Control of ion loading into the xylem has been repeatedly named as a crucial factor determining plant salt tolerance. In this study we further investigate this issue by applying a range of biophysical [the microelectrode ion flux measurement (MIFE) technique for non-invasive ion flux measurements, the patch clamp technique, membrane potential measurements] and physiological (xylem sap and tissue nutrient analysis, photosynthetic characteristics, stomatal conductance) techniques to barley varieties contrasting in their salt tolerance. We report that restricting Na(+) loading into the xylem is not essential for conferring salinity tolerance in barley, with tolerant varieties showing xylem Na(+) concentrations at least as high as those of sensitive ones. At the same time, tolerant genotypes are capable of maintaining higher xylem K(+)/Na(+) ratios and efficiently sequester the accumulated Na(+) in leaves. The former is achieved by more efficient loading of K(+) into the xylem. We argue that the observed increases in xylem K(+) and Na(+) concentrations in tolerant genotypes are required for efficient osmotic adjustment, needed to support leaf expansion growth. We also provide evidence that K(+)-permeable voltage-sensitive channels are involved in xylem loading and operate in a feedback manner to maintain a constant K(+)/Na(+) ratio in the xylem sap. PMID- 20015064 TI - An efficient system to detect protein ubiquitination by agroinfiltration in Nicotiana benthamiana. AB - The ubiquitination proteasome pathway has been demonstrated to regulate all plant developmental and signaling processes. E3 ligase/substrate-specific interactions and ubiquitination play important roles in this pathway. However, due to technical limitations only a few instances of E3 ligase-substrate binding and protein ubiquitination in plants have been directly evidenced. An efficient in vivo and in vitro ubiquitination assay was developed for analysis of protein ubiquitination reactions by agroinfiltration expression of both substrates and E3 ligases in Nicotiana benthamiana. Using a detailed analysis of the well-known E3 ligase COP1 and its substrate HY5, we demonstrated that this assay allows for fast and reliable detection of the specific interaction between the substrate and the E3 ligase, as well as the effects of MG132 and substrate ubiquitination and degradation. We were able to differentiate between the original and ubiquitinated forms of the substrate in vivo with antibodies to ubiquitin or to the target protein. We also demonstrated that the substrate and E3 ligase proteins expressed by agroinfiltration can be applied to analyze ubiquitination in in vivo or in vitro reactions. In addition, we optimized the conditions for different types of substrate and E3 ligase expression by supplementation with the gene-silencing suppressor p19 and by time-courses of sample collection. Finally, by testing different protein extraction buffers, we found that different types of buffer should be used for different ubiquitination analyses. This method should be adaptable to other protein modification studies. PMID- 20015065 TI - Dynamic regulation of guard cell anion channels by cytosolic free Ca2+ concentration and protein phosphorylation. AB - In guard cells, activation of anion channels (I(anion)) is an early event leading to stomatal closure. Activation of I(anion) has been associated with abscisic acid (ABA) and its elevation of the cytosolic free Ca(2+) concentration ([Ca(2+)](i)). However, the dynamics of the action of [Ca(2+)](i) on I(anion) has never been established, despite its importance for understanding the mechanics of stomatal adaptation to stress. We have quantified the [Ca(2+)](i) dynamics of I(anion) in Vicia faba guard cells, measuring channel current under a voltage clamp while manipulating and recording [Ca(2+)](i) using Fura-2 fluorescence imaging. We found that I(anion) rises with [Ca(2+)](i) only at concentrations substantially above the mean resting value of 125 +/- 13 nm, yielding an apparent K(d) of 720 +/- 65 nm and a Hill coefficient consistent with the binding of three to four Ca(2+) ions to activate the channels. Approximately 30% of guard cells exhibited a baseline of I(anion) activity, but without a dependence of the current on [Ca(2+)](i). The protein phosphatase antagonist okadaic acid increased this current baseline over twofold. Additionally, okadaic acid altered the [Ca(2+)](i) sensitivity of I(anion), displacing the apparent K(d) for [Ca(2+)](i) to 573 +/- 38 nm. These findings support previous evidence for different modes of regulation for I(anion), only one of which depends on [Ca(2+)](i), and they underscore an independence of [Ca(2+)](i) from protein (de-)phosphorylation in controlling I(anion). Most importantly, our results demonstrate a significant displacement of I(anion) sensitivity to higher [Ca(2+)](i) compared with that of the guard cell K(+) channels, implying a capacity for variable dynamics between net osmotic solute uptake and loss. PMID- 20015067 TI - Induced resistance to pests and pathogens in trees. AB - Tree resistance can be enhanced by a variety of biotic and abiotic inducers, including nonpathogenic and pathogenic microbes, and herbivores, resulting in enhanced protection against further biotic injury. Induced resistance (IR) could be a valuable tool in sustainable pest management. IR has been actively studied in herbaceous plant species, and, in recent years, in woody plant species, and is fast emerging as an intriguing, eco-friendly concept for enhancing tree resistance. However, before application of IR becomes possible, there is a need to increase our knowledge of the mechanisms of defence in forest trees. A richer understanding of these phenomena will play a critical role in developing sustainable integrated pest management strategies. This review summarizes our current knowledge of IR in forest trees, focusing on inducible defence mechanisms, systemic induction of resistance and phytohormone signalling networks. We conclude by discussing the potential advantages and limitations of applying IR-based management tools in forest systems. PMID- 20015066 TI - Adaptation of Medicago truncatula to nitrogen limitation is modulated via local and systemic nodule developmental responses. AB - Adaptation of Medicago truncatula to local nitrogen (N) limitation was investigated to provide new insights into local and systemic N signaling. The split-root technique allowed a characterization of the local and systemic responses of NO(3)(-) or N(2)-fed plants to localized N limitation. (15)N and (13)C labeling were used to monitor plant nutrition. Plants expressing pMtENOD11 GUS and the sunn-2 hypernodulating mutant were used to unravel mechanisms involved in these responses. Unlike NO(3)(-)-fed plants, N(2)-fixing plants lacked the ability to compensate rapidly for a localized N limitation by up regulating the N(2)-fixation activity of roots supplied elsewhere with N. However they displayed a long-term response via a growth stimulation of pre-existing nodules, and the generation of new nodules, likely through a decreased abortion rate of early nodulation events. Both these responses involve systemic signaling. The latter response is abolished in the sunn mutant, but the mutation does not prevent the first response. Local but also systemic regulatory mechanisms related to plant N status regulate de novo nodule development in Mt, and SUNN is required for this systemic regulation. By contrast, the stimulation of nodule growth triggered by systemic N signaling does not involve SUNN, indicating SUNN independent signaling. PMID- 20015068 TI - Post-translational protein modification as a tool for transcription reprogramming. AB - Precise modulation of transcription plays a vital role in both development and the response of all higher organisms to their environment. Temporal activation or repression of specific genes is accomplished via a plethora of transcriptional regulators. However, relatively little is known about how the activities of these proteins are controlled. Recent findings indicate that post-translational modifications fine-tune the function of transcription regulators by affecting their localization, conformation or stability. Here, we discuss these regulatory mechanisms in the context of the plant immune response. This system lends itself particularly well to studies of transcriptional regulators as activation of plant immunity is associated with rapid and dramatic reprogramming of the transcriptome. A case study of the plant immune coactivator NPR1 (nonexpressor of pathogenesis-related (PR) genes 1) illustrates that transcription regulator activity may be controlled by redox-based modifications of cysteine thiols (e.g. disulphide bonding and S-nitrosylation), phosphorylation, and ubiquitinylation coupled to protein degradation. Importantly, cross-talk between distinct protein modifications may determine the spatial and temporal activity of transcription regulators that in turn profile the cellular transcriptome. PMID- 20015069 TI - Sesbania rostrata: a case study of natural variation in legume nodulation. AB - Legumes acquired the ability to engage in a symbiotic interaction with soil-borne bacteria and establish a nitrogen-fixing symbiosis in a novel root organ, the nodule. Most legume crops and the model legumes Medicago truncatula and Lotus japonicus are infected intracellularly in root hairs via infection threads that lead the bacteria towards a nodule primordium in the root cortex. This infection process, however, does not reflect the great diversity of infection strategies that are used by leguminous plants. An alternative, intercellular invasion occurs in the semiaquatic legume Sesbania rostrata. Bacteria colonize epidermal fissures at lateral root bases and trigger cortical cell death for infection pocket formation and subsequent intercellular and intracellular infection thread progression towards the primordium. This infection mode evolved as an adaptation to waterlogged conditions that inhibit intracellular invasion. In this review, we discuss the molecular basis for this adaptation and how insights into this process contribute to general knowledge of the rhizobial infection process. PMID- 20015070 TI - Digging deeper: fine-root responses to rising atmospheric CO concentration in forested ecosystems. AB - Experimental evidence from a diverse set of forested ecosystems indicates that CO2 enrichment may lead to deeper rooting distributions. While the causes of greater root production at deeper soil depths under elevated CO2 concentration ([CO2]) require further investigation, altered rooting distributions are expected to affect important ecosystem processes. The depth at which fine roots are produced may influence root chemistry, physiological function, and mycorrhizal infection, leading to altered nitrogen (N) uptake rates and slower turnover. Also, soil processes such as microbial decomposition are slowed at depth in the soil, potentially affecting the rate at which root detritus becomes incorporated into soil organic matter. Deeper rooting distributions under elevated [CO2] provide exciting opportunities to use novel sensors and chemical analyses throughout the soil profile to track the effects of root proliferation on carbon (C) and N cycling. Models do not currently incorporate information on root turnover and C and N cycling at depth in the soil, and modification is necessary to accurately represent processes associated with altered rooting depth distributions. Progress in understanding and modeling the interface between deeper rooting distributions under elevated [CO2] and soil C and N cycling will be critical in projecting the sustainability of forest responses to rising atmospheric [CO2]. PMID- 20015071 TI - The central role of CDE/CHR promoter elements in the regulation of cell cycle dependent gene transcription. AB - The cell cycle-dependent element (CDE) and the cell cycle genes homology region (CHR) control the transcription of genes with maximum expression in G(2) phase and in mitosis. Promoters of these genes are repressed by proteins binding to CDE/CHR elements in G(0) and G(1) phases. Relief from repression begins in S phase and continues into G(2) phase and mitosis. Generally, CDE sites are located four nucleotides upstream of CHR elements in TATA-less promoters of genes such as Cdc25C, Cdc2 and cyclin A. However, expression of some other genes, such as human cyclin B1 and cyclin B2, has been shown to be controlled only by a CHR lacking a functional CDE. To date, it is not fully understood which proteins bind to and control CDE/CHR-containing promoters. Recently, components of the DREAM complex were shown to be involved in CDE/CHR-dependent transcriptional regulation. In addition, the expression of genes regulated by CDE/CHR elements is mostly achieved through CCAAT-boxes, which bind heterotrimeric NF-Y proteins as well as the histone acetyltransferase p300. Importantly, many CDE/CHR promoters are downregulated by the tumor suppressor p53. In this review, we define criteria for CDE/CHR-regulated promoters and propose to distinguish two classes of CDE/CHR regulated genes. The regulation through transcription factors potentially binding to the CDE/CHR is discussed, and recently discovered links to central pathways regulated by E2F, the pRB family and p53 are highlighted. PMID- 20015072 TI - Enzymatic and electron paramagnetic resonance studies of anabolic pyruvate synthesis by pyruvate: ferredoxin oxidoreductase from Hydrogenobacter thermophilus. AB - Pyruvate: ferredoxin oxidoreductase (POR; EC 1.2.7.1) catalyzes the thiamine pyrophosphate-dependent oxidative decarboxylation of pyruvate to form acetyl-CoA and CO(2). The thermophilic, obligate chemolithoautotrophic hydrogen-oxidizing bacterium, Hydrogenobacter thermophilus TK-6, assimilates CO(2) via the reductive tricarboxylic acid cycle. In this cycle, POR acts as pyruvate synthase catalyzing the reverse reaction (i.e. reductive carboxylation of acetyl-CoA) to form pyruvate. The pyruvate synthesis reaction catalyzed by POR is an energetically unfavorable reaction and requires a strong reductant. Moreover, the reducing equivalents must be supplied via its physiological electron mediator, a small iron-sulfur protein ferredoxin. Therefore, the reaction is difficult to demonstrate in vitro and the reaction mechanism has been poorly understood. In the present study, we coupled the decarboxylation of 2-oxoglutarate catalyzed by 2-oxoglutarate: ferredoxin oxidoreductase (EC 1.2.7.3), which generates sufficiently low-potential electrons to reduce ferredoxin, to drive the energy demanding pyruvate synthesis by POR. We demonstrate that H. thermophilus POR catalyzes pyruvate synthesis from acetyl-CoA and CO(2), confirming the operation of the reductive tricarboxylic acid cycle in this bacterium. We also measured the electron paramagnetic resonance spectra of the POR intermediates in both the forward and reverse reactions, and demonstrate the intermediacy of a 2-(1 hydroxyethyl)- or 2-(1-hydroxyethylidene)-thiamine pyrophosphate radical in both reactions. The reaction mechanism of the reductive carboxylation of acetyl-CoA is also discussed. PMID- 20015073 TI - Application of the Fc fusion format to generate tag-free bi-specific diabodies. AB - We previously reported the use of a humanized bi-specific diabody that targets epidermal growth factor receptor and CD3 (hEx3-Db) for cancer immunotherapy. Bacterial expression can be used to express small recombinant antibodies on a large scale; however, their overexpression often results in the formation of insoluble aggregates, and in most cases artificial affinity peptide tags need to be fused to the antibodies for purification by affinity chromatography. Here, we propose a novel method for preparing refined, functional, tag-free bi-specific diabodies from IgG-like bi-specific antibodies (BsAbs) in a mammalian expression system. We created an IgG-like BsAb in which bi-specific diabodies were fused to the human Fc region via a designed human rhinovirus 3C (HRV3C) protease recognition site. The BsAb was purified by protein A affinity chromatography, and the refined tag-free hEx3-Db was efficiently produced from the Fc fusion format by protease digestion. The tag-free hEx3-Db from the Fc fusion format showed a greater inhibition of cancer growth than affinity-tagged hEx3-Db prepared directly from Chinese hamster ovary cells. We also applied our novel method to another small recombinant antibody fragment, hEx3 single-chain diabody (hEx3 scDb), and demonstrated the versatility and advantages of our proposed method compared with papain digestion of hEx3-scDb. This approach may be used for industrial-scale production of functional tag-free small therapeutic antibodies. PMID- 20015074 TI - The myostatin-induced E3 ubiquitin ligase RNF13 negatively regulates the proliferation of chicken myoblasts. AB - The ubiquitin ligase RING finger protein 13 gene (RNF13) was first identified in a screen for genes whose expression is regulated by myostatin in chicken fetal myoblasts. In this study, we demonstrate that the RNF13 gene is broadly expressed in many chicken tissues. The expression of RNF13 gradually decreases during skeletal myogenesis, and myostatin up-regulates RNF13 expression at both the transcriptional and translational levels. Interestingly, ectopic expression of RNF13 inhibits cell proliferation and suppresses the expression of the myogenic genes MyoD and Caveolin-3 in muscle cells. Moreover, recently, we have reported that RNF13 is a RING-type E3 ubiquitin ligase. In this report, we provide experimental evidence to show that mutations disrupting the RING finger abolish the growth-suppressive activity of RNF13, indicating that its E3 ligase activity is required for the inhibition of cell proliferation. Taken together, our findings show that RNF13 functions as an E3 ubiquitin ligase to negatively regulate cell proliferation. PMID- 20015076 TI - Novel L-amino acid oxidase with antibacterial activity against methicillin resistant Staphylococcus aureus isolated from epidermal mucus of the flounder Platichthys stellatus. AB - Fish produce mucus substances as a defensive outer barrier against environmental xenobiotics and predators. Recently, we found a bioactive protein in the mucus layer of the flounder Platichthys stellatus, which showed antibacterial activity against Staphylococcus epidermidis, Staphylococcus aureus and methicillin resistant S. aureus. In this study, we isolated and identified the antibacterial protein from the mucus components of P. stellatus using a series of column chromatography steps. We then performed gel electrophoresis and cDNA cloning to characterize the protein. The antibacterial protein in the mucus had a molecular mass of approximately 52 kDa with an isoelectric point of 5.3, and cDNA sequencing showed that it corresponded completely with the peptide sequence of antibacterial protein from the gill. A BLAST search suggested that the cDNA encoded an antibacterial protein sharing identity with a number of L-amino acid oxidases (LAAOs) and possessing several conserved motifs found in flavoproteins. RT-PCR using a specific primer, and immunohistochemical analysis with anti-LAAO IgG, demonstrated tissue-specific expression and localization in the gill. Moreover, the anti-LAAO IgG was able to neutralize the antibacterial activity of the protein against methicillin-resistant S. aureus. Thus, we demonstrated that this antibacterial protein, identified from P. stellatus-derived epidermal mucus, is a novel LAAO-like protein with antibacterial activity, similar to snake LAAOs. PMID- 20015077 TI - Enhanced sensitivity to hydrogen peroxide-induced apoptosis in Evi1 transformed Rat1 fibroblasts due to repression of carbonic anhydrase III. AB - EVI1 is a nuclear zinc finger protein essential to normal development, which participates in acute myeloid leukaemia progression and transforms Rat1 fibroblasts. In this study we show that enforced expression of Evi1 in Rat1 fibroblasts protects from paclitaxel-induced apoptosis, consistent with previously published studies. Surprisingly, however, these cells show increased sensitivity to hydrogen peroxide (H(2)O(2))-induced apoptosis, demonstrated by elevated caspase 3 catalytic activity. This effect is caused by a reduction in carbonic anhydrase III (caIII) production. caIII transcripts are repressed by 92 97% by Evi1 expression, accompanied by a similar reduction in caIII protein. Reporter assays with the rat caIII gene promoter show repressed activity, demonstrating that Evi1 either directly or indirectly modulates transcription of this gene in Rat1 cells. Targeted knockdown of caIII alone, with Dicer-substrate short inhibitory RNAs, also increases the sensitivity of Rat1 fibroblasts to H(2)O(2), which occurs in the absence of any other changes mediated by Evi1 expression. Enforced expression of caIII in Evi1-expressing Rat1 cells reverts the phenotype, restoring H(2)O(2) resistance. Together these data show that Evi1 represses transcription of caIII gene expression, leading to increased sensitivity to H(2)O(2)-induced apoptosis in Rat1 cells and might suggest the basis for the development of a novel therapeutic strategy for the treatment of leukaemias and solid tumours where EVI1 is overexpressed. PMID- 20015075 TI - Aegyptin displays high-affinity for the von Willebrand factor binding site (RGQOGVMGF) in collagen and inhibits carotid thrombus formation in vivo. AB - Aegyptin is a 30 kDa mosquito salivary gland protein that binds to collagen and inhibits platelet aggregation. We have studied the biophysical properties of aegyptin and its mechanism of action. Light-scattering plot showed that aegyptin has an elongated monomeric form, which explains the apparent molecular mass of 110 kDa estimated by gel-filtration chromatography. Surface plasmon resonance identified the sequence RGQOGVMGF (where O is hydroxyproline) that mediates collagen interaction with von Willebrand factor (vWF) as a high-affinity binding site for aegyptin, with a K(D) of approximately 5 nM. Additionally, aegyptin interacts with the linear peptide RGQPGVMGF and heat-denatured collagen, indicating that the triple helix and hydroxyproline are not a prerequisite for binding. However, aegyptin does not interact with scrambled RGQPGVMGF peptide. Aegyptin also recognizes the peptides (GPO)(10) and GFOGER with low affinity (microM range), which respectively represent glycoprotein VI and integrin alpha2beta1 binding sites in collagen. Truncated forms of aegyptin were engineered, and the C-terminus fragment was shown to interact with collagen and to attenuate platelet aggregation. In addition, aegyptin prevents laser-induced carotid thrombus formation in the presence of Rose Bengal in vivo, without significant bleeding in rats. In conclusion, aegyptin interacts with distinct binding sites in collagen, and is useful tool to inhibit platelet-collagen interaction in vitro and in vivo. PMID- 20015078 TI - From meiosis to postmeiotic events: the secrets of histone disappearance. AB - One of the most obscure phenomena in modern biology is the near genome-wide displacement of histones that occurs during the postmeiotic phases of spermatogenesis in many species. Here we review the literature to show that, during spermatogenic differentiation, three major molecular mechanisms come together to 'prepare' the nucleosomes for facilitated disassembly and histone removal. PMID- 20015080 TI - From meiosis to postmeiotic events: homologous recombination is obligatory but flexible. AB - Sexual reproduction depends on the success of faithful chromosome transmission during meiosis to yield viable gametes. Central to meiosis is the process of recombination between paternal and maternal chromosomes, which boosts the genetic diversity of progeny and ensures normal homologous chromosome segregation. Imperfections in meiotic recombination are the source of de novo germline mutations, abnormal gametes, and infertility. Thus, not surprisingly, cells have developed a variety of mechanisms and tight controls to ensure sufficient and well-distributed recombination events within their genomes, the details of which remain to be fully elucidated. Local and genome-wide studies of normal and genetically engineered cells have uncovered a remarkable stochasticity in the number and positioning of recombination events per chromosome and per cell, which reveals an impressive level of flexibility. In this minireview, we summarize our contemporary understanding of meiotic recombination and its control mechanisms, and address the seemingly paradoxical and poorly understood diversity of recombination sites. Flexibility in the distribution of meiotic recombination events within genomes may reside in regulation at the chromatin level, with histone modifications playing a recently recognized role. PMID- 20015081 TI - From meiosis to postmeiotic events: alignment and recognition of homologous chromosomes in meiosis. AB - Recombination of homologous chromosomes is essential for correct reductional segregation of homologous chromosomes, which characterizes meiosis. To accomplish homologous recombination, chromosomes must find their homologous partners and pair with them within the spatial constraints of the nucleus. Although various mechanisms have developed in different organisms, two major steps are involved in the process of pairing: first, alignment of homologous chromosomes to bring them close to each other for recognition; and second, recognition of the homologous partner of each chromosome so that they can form an intimate pair. Here, we discuss the various mechanisms used for alignment and recognition of homologous chromosomes in meiosis. PMID- 20015079 TI - From meiosis to postmeiotic events: uncovering the molecular roles of the meiosis specific recombinase Dmc1. AB - In meiosis, the accurate segregation of maternal and paternal chromosomes is accomplished by homologous recombination. A central player in meiotic recombination is the Dmc1 recombinase, a member of the RecA/Rad51 recombinase superfamily, which is widely conserved from viruses to humans. Dmc1 is a meiosis specific protein that functions with the ubiquitously expressed homolog, the Rad51 recombinase, which is essential for both mitotic and meiotic recombination. Since its discovery, it has been speculated that Dmc1 is important for unique aspects of meiotic recombination. Understanding the distinctive properties of Dmc1, namely, the features that distinguish it from Rad51, will further clarify the mechanisms of meiotic recombination. Recent structural, biochemical, and genetic findings are now revealing the molecular mechanisms of Dmc1-mediated homologous recombination and its regulation by various recombination mediators. PMID- 20015082 TI - Dynamics in the transmission of genetic information: from meiosis to postmeiotic events. PMID- 20015083 TI - Is anticonvulsant treatment of mania a class effect? Data from randomized clinical trials. AB - Our aim was to evaluate the efficacy and tolerability of anticonvulsant agents for the treatment of acute bipolar mania and ascertain if their effects on mania are a "class" effect. We conducted a systematic review of randomized controlled trials (RCTs) with placebo or active comparator, in acute bipolar mania in order to summarize available data on anticonvulsant treatment of mania/mixed episodes. We searched (PubMed/MEDLINE) with the combination of the words "acute mania" and "clinical trials" with each one of the following words: "anticonvulsants/antiepileptics,""valproic/valproate/divalproex,""carbamazepine," oxcarbazepine,""lamotrigine,""gabapentin,""topiramate,""phenytoin,""zonisamide,"" etigabine,""pregabalin,""tiagabine,""levetiracetam,""licarbazepine,""felbamate," and "vigabatrin." Original articles were found until November 1, 2008. Data from 35 randomized clinical trials suggested that not all anticonvulsants are efficacious for the treatment of acute mania. Valproate showed greater efficacy in reducing manic symptoms, with response rates around 50% compared to a placebo effect of 20-30%. It appears to have a more robust antimanic effect than lithium in rapid cycling and mixed episodes. As valproate, the antimanic effects of carbamazepine have been demonstrated. Evidences did not support the efficacy of the gabapentin, topiramate as well as lamotrigine as monotherapy in acute mania and mixed episodes. Oxcarbazepine data are inconclusive and data regarding other anticonvulsants are not available. Anticonvulsants are not a class when treating mania. While valproate and carbamazepine are significantly more effective than placebo, gabapentin, topiramate, and lamotrigine are not. However, some anticonvulsants may be efficacious in treating some psychiatric comorbidities that are commonly associated to bipolar illness. PMID- 20015084 TI - Effect of long- and short-term treatments with pravastatin on diabetes mellitus and pancreatic fibrosis in the Otsuka-Long-Evans-Tokushima fatty rat. AB - BACKGROUND AND PURPOSE: The effects of statins on diabetes mellitus (DM) are controversial, and their effects on pancreatic fibrosis are poorly defined. We investigated the effect of long- and short-term treatments with pravastatin on the development of DM and pancreatic fibrosis in DM-prone Otsuka-Long-Evans Tokushima Fatty (OLETF) rats. EXPERIMENTAL APPROACH: Male OLETF rats were divided into four groups at 12 weeks of age. The first group received a standard rat diet until the end of the experimental period at age 80 weeks. The second group was given a diet containing 0.05% pravastatin from 12 weeks of age, before the onset of DM and pancreatic fibrosis, and the third group was given the same pravastatin diet from 28 weeks of age, after the onset of DM and pancreatic fibrosis, until age 80 weeks. The fourth group received the same pravastatin diet only for 16 weeks, from 12 to 28 weeks of age, and switched to a standard diet. Progressions of DM and pancreatic fibrosis were evaluated. KEY RESULTS: Long-term treatments with pravastatin, either from 12 or 28 weeks of age, decreased serum glucose concentration and fibrotic area, elevated superoxide dismutase activity and down regulated transforming growth factor-beta1 mRNA in the pancreas. In contrast, after a short-term treatment with pravastatin, these parameters markedly deteriorated after its cessation. CONCLUSIONS AND IMPLICATIONS: The results suggest that long-term treatment with pravastatin improves DM and pancreatic fibrosis via anti-oxidative and anti-fibrotic properties, whereas cessation of pravastatin abolishes these beneficial effects, and accelerates DM and pancreatic fibrosis. PMID- 20015085 TI - Effects of chronic sympatho-inhibition on reflex control of renal blood flow and plasma renin activity in renovascular hypertension. AB - BACKGROUND AND PURPOSE: We determined if chronic sympatho-inhibition with rilmenidine has functional significance for the kidney by altering responses of renal blood flow (RBF) and plasma renin activity (PRA) to stress and acute hypotension in rabbits with renovascular hypertension. EXPERIMENTAL APPROACH: RBF to each kidney and renal sympathetic nerve activity (RSNA) to the left kidney were measured in rabbits in which a renal artery clip induced hypertension (2K1C) and in sham-operated rabbits. After 2 weeks, a subcutaneous minipump was implanted to deliver rilmenidine (2.5 mg.kg(-1).day(-1)) to 2K1C rabbits for 3 weeks. KEY RESULTS: After 5 weeks of renal artery stenosis, mean arterial pressure (MAP) was 23% higher and PRA 3-fold greater than in sham-operated rabbits. Blood flow and renal vascular conductance in the stenosed kidney were lower (-75% and -80%) compared with sham, and higher in the non-clipped kidney (68% and 39%). Responses of RBF and PRA to hypotension were similar in 2K1C and sham rabbits. Airjet stress evoked a greater increase in MAP in 2K1C rabbits than sham controls. Chronic rilmenidine normalized MAP, reduced RSNA and PRA, and did not reduce RBF in the stenosed kidney. Responses of RBF (clipped and non-clipped kidney), RSNA and PRA to hypotension and airjet were little affected by rilmenidine. CONCLUSIONS AND IMPLICATIONS: Our observations suggest that chronic sympatho-inhibition is an effective antihypertensive therapy in renovascular hypertension. It normalizes MAP and reduces basal PRA without compromising blood flow in the stenosed kidney or altering responses of MAP, haemodynamics and PRA to acute hypotension and stress. PMID- 20015086 TI - A unique modulator of endoplasmic reticulum stress-signalling pathways: the novel pharmacological properties of amiloride in glial cells. AB - BACKGROUND AND PURPOSE: Stress on the endoplasmic reticulum (ER) can trigger rescuer responses such as the unfolded protein response (UPR). However, pharmacological modulators of these ER-regulated stress responses are not well understood. In the present study, we found that amiloride, a potassium-sparing diuretic, has unique properties relating to such stress. EXPERIMENTAL APPROACH: We treated mouse primary cultured glial cells with amiloride, in the absence and presence of the ER stress-inducing reagents tunicamycin (Tm) or dithiothreitol, and measured UPR and ER stress-induced cell death. IRE1alpha phosphorylation, eIF2alpha phosphorylation, X-box binding protein 1 (XBP1) splicing, glucose regulated protein 78 (GRP78) and CCAAT/enhancer-binding protein homologous protein (CHOP) expression by reverse transcription-polymerase chain reaction and Western blotting were used to assess UPR and lactate dehydrogenase activity was determined to measure ER stress-induced cell death. KEY RESULTS: Amiloride completely inhibited ER stress-induced activation of IRE1alpha, an ER-localized stress sensor protein, splicing of XBP1, and subsequent expression of GRP78 at the mRNA and protein levels. ER stress induces the phosphorylation of eIF2alpha, leading to the expression of CHOP or an attenuation of translation in cells. Surprisingly, treatment with amiloride alone markedly promoted the phosphorylation but actually inhibited ER stress-induced CHOP expression. Finally, we found that amiloride (200 microM) synergistically enhanced ER stress induced cell death, which was mediated through caspases. On the other hand, a low dose of amiloride (20 microM) significantly prevented Tm-induced cell death. CONCLUSIONS AND IMPLICATIONS: These results suggest that amiloride can modulate UPR. They also suggest amiloride to be an important pharmacological agent and provide basic information for understanding and preventing ER stress-related diseases. PMID- 20015087 TI - The distribution of saponins in vivo affects their synergy with chimeric toxins against tumours expressing human epidermal growth factor receptors in mice. AB - BACKGROUND AND PURPOSE: Certain saponins synergize with antitumour drugs to enhance their efficacy, but the mechanisms underlying this synergy in vivo are not well studied. Here, we describe the distribution of Saponinum album (Spn) from Gypsophila paniculata L. in mice after subcutaneous injection. EXPERIMENTAL APPROACH: The [(3)H]-labelled Spn used for in vivo experiments was biologically active, as it still increased the cytotoxicity of a chimeric toxin in vitro. Distribution of [(3)H]-Spn was measured in BALB/c mice, with or without subcutaneous tumours in the flank. Labelled Spn was subcutaneously injected in the neck, and samples of organs, blood, urine and tumour tissue were analysed for radioactivity, 5-240 min after the injection. KEY RESULTS: The majority of [(3)H] Spn distributed within 10 min throughout the entire animal, with high levels of radioactivity in the urine by 30 min. No preferential accumulation in tumour tissue or other organs was observed. In tumour-bearing mice, using a sequential combination of Spn (given first) and a chimeric toxin against the epidermal growth factor receptor, ErbB1, we tested two different pretreatment times for Spn. There was high antitumour efficacy (66% inhibition of tumour growth) after 60 min pre treatment with Spn, but no significant inhibition after 10 min pre treatment with Spn. CONCLUSIONS AND IMPLICATIONS: [(3)H]-Spn was rapidly cleared from the mice after s.c. injection, and antitumour synergy with chimeric toxins was correlated with the removal of excess Spn from tissues. Disposition of Spn in vivo may critically determine antitumour synergy with chimeric toxins. PMID- 20015088 TI - Mechanisms of Kv2.1 channel inhibition by celecoxib--modification of gating and channel block. AB - BACKGROUND AND PURPOSE: Selective cyclooxygenase-2 (COX-2) inhibitors such as rofecoxib (Vioxx) and celecoxib (Celebrex) were developed as NSAIDs with reduced gastric side effects. Celecoxib has now been shown to affect cellular physiology via an unexpected, COX-independent, pathway - by inhibiting K(v)2.1 and other ion channels. In this study, we investigated the mechanism of the action of celecoxib on K(v)2.1 channels. EXPERIMENTAL APPROACH: The mode of action of celecoxib on rat K(v)2.1 channels was studied by whole-cell patch-clamping to record currents from channels expressed in HEK-293 cells. KEY RESULTS: Celecoxib reduced current through K(v)2.1 channels when applied from the extracellular side. At low concentrations (3 microM, celecoxib led to closed-channel block with relative slowing of activation. At 30 microM, it additionally induced open-channel block that manifested in use-dependent inhibition and slower recovery from inactivation. CONCLUSIONS AND IMPLICATIONS: Celecoxib reduced current through K(v)2.1 channels by modifying gating and inducing closed- and open-channel block, with the three effects manifesting at different concentrations. These data will help to elucidate the mechanisms of action of this widely prescribed drug on ion channels and those underlying its neurological, cardiovascular and other effects. PMID- 20015089 TI - The sphingosine kinase inhibitor N,N-dimethylsphingosine inhibits neointimal hyperplasia. AB - BACKGROUND AND PURPOSE: Sphingosine-1-phosphate and its receptors may be involved in vascular smooth muscle cell (VSMC) proliferation following vascular injury. Here, we evaluate the effect of d-erythro-N,N-dimethylsphingosine (DMS), a sphingosine kinase (SK) inhibitor, on VSMC proliferation, apoptosis and neointimal formation. EXPERIMENTAL APPROACH: Growth responses in vitro to fetal calf serum (FCS) were measured by [(3)H]-thymidine incorporation and extracellular signal-regulated kinase-1/2 (ERK-1/2) activation in quiescent primary cultures of porcine VSMC in the presence and absence of various concentrations of the SK inhibitor DMS. In vivo treatment with DMS was delivered with a local endoluminal catheter, following balloon injury of coronary arteries. The artery intimal formation was investigated by angiography, myography and histomorphometry. KEY RESULTS: In vitro experiments indicated that DMS induced a dose-dependent reduction in [(3)H]-thymidine incorporation and ERK-1/2 activation via a protein kinase C (PKC) independent mechanism with an IC(50) value of 12 +/- 6 and 15 +/- 10 microM respectively. DMS also reduced Akt signalling. Four weeks following in vivo delivery of DMS, complete functional endothelial regeneration was observed in all treatment groups, with significant reduction of intimal formation (vehicle 23.7 +/- 4.6% vs. DMS infusion 8.92 +/- 2.9%, P < 0.05). CONCLUSIONS AND IMPLICATIONS: Taken together, these results demonstrate that local administration of the SK inhibitor, DMS, reduced neointimal formation, and this effect could involve inhibition of ERK-1/2 and Akt signalling, and modulation of smooth muscle growth. PMID- 20015090 TI - Isolation and pharmacological characterization of AdTx1, a natural peptide displaying specific insurmountable antagonism of the alpha1A-adrenoceptor. AB - BACKGROUND AND PURPOSE: Venoms are a rich source of ligands for ion channels, but very little is known about their capacity to modulate G-protein coupled receptor (GPCR) activity. We developed a strategy to identify novel toxins targeting GPCRs. EXPERIMENTAL APPROACH: We studied the interactions of mamba venom fractions with alpha(1)-adrenoceptors in binding experiments with (3)H-prazosin. The active peptide (AdTx1) was sequenced by Edman degradation and mass spectrometry fragmentation. Its synthetic homologue was pharmacologically characterized by binding experiments using cloned receptors and by functional experiments on rabbit isolated prostatic smooth muscle. KEY RESULTS: AdTx1, a 65 amino-acid peptide stabilized by four disulphide bridges, belongs to the three finger-fold peptide family. It has subnanomolar affinity (K(i)= 0.35 nM) and high specificity for the human alpha(1A)-adrenoceptor subtype. We showed high selectivity and affinity (K(d)= 0.6 nM) of radio-labelled AdTx1 in direct binding experiments and revealed a slow association constant (k(on)= 6 x 10(6).M(-1).min( 1)) with an unusually stable alpha(1A)-adrenoceptor/AdTx1 complex (t(1/2diss)= 3.6 h). AdTx1 displayed potent insurmountable antagonism of phenylephrine's actions in vitro (rabbit isolated prostatic muscle) at concentrations of 10 to 100 nM. CONCLUSIONS AND IMPLICATIONS: AdTx1 is the most specific and selective peptide inhibitor for the alpha(1A)-adrenoceptor identified to date. It displays insurmountable antagonism, acting as a potent relaxant of smooth muscle. Its peptidic nature can be exploited to develop new tools, as a radio-labelled-AdTx1 or a fluoro-labelled-AdTx1. Identification of AdTx1 thus offers new perspectives for developing new drugs for treating benign prostatic hyperplasia. PMID- 20015092 TI - The methyl ester of okadaic acid is more potent than okadaic acid in disrupting the actin cytoskeleton and metabolism of primary cultured hepatocytes. AB - BACKGROUND AND PURPOSE: Okadaic acid (OA) and microcystins (MCs) are structurally different toxins with the same mechanism of action, inhibition of serine/threonine protein phosphatases (PPs). Methyl okadaate (MeOk), a methyl ester derivative of OA, was considered almost inactive due to its weak inhibition of PP1 and PP2A. Here, we have investigated the activity and potency of MeOk in hepatic cells in comparison with that of OA and MCs. EXPERIMENTAL APPROACH: We tested the effects of MeOK, OA and microcystin-leucine and arginine (MC-LR) on the metabolic rate, the actin cytoskeleton and glucose uptake in a rat hepatocyte cell line (Clone 9) and in primary cultured rat hepatocytes. PP2A was assayed to compare OA and MeOk activity. KEY RESULTS: MeOk disrupted the actin cytoskeleton and depressed the metabolic rate of both types of rat hepatocytes, being six-fold less potent than OA in Clone 9 cells but nearly six-fold more potent in primary cultured hepatocytes. However, unlike OA, MeOk did not change glucose uptake in these cells, suggesting a weak inhibition of PP2A, as confirmed in direct assays of PP2A activity. CONCLUSIONS AND IMPLICATIONS: Although MeOk was originally described as a weakly bioactive molecule, it clearly depressed the metabolic rate and disrupted the cytoskeleton in primary and immortalized rat hepatocytes. Furthermore, MeOk affected primary hepatocytes at much lower concentrations than those affecting immortalized cells. These effects were unrelated to PP2A inhibition. Our results suggest the risk to public health from MeOk in foodstuffs should be re-evaluated. PMID- 20015093 TI - Decreased expression of Fli-1 in bone marrow-derived haematopoietic cells significantly affects disease development in Murphy Roths Large/lymphoproliferation (MRL/lpr) mice. AB - The transcription factor Fli-1 is implicated in the pathogenesis of both murine and human lupus. Decreased expression of Fli-1 in heterozygous (Fli-1(+/-)) Murphy Roths Large (MRL)/lpr mice resulted in significantly lower kidney pathological scores and markedly increased survival. In this study, bone marrow (BM) transplantation was used to investigate the role of decreased expression of Fli-1 in haematopoietic versus non-haematopoietic cell lineages in autoimmune disease development. Wild-type (WT) MRL/lpr that received BM from Fli-1(+/-) MRL/lpr mice had statistically significantly lower autoantibodies, less proteinuria, reduced renal disease and prolonged survival compared to WT MRL/lpr mice that received BM from WT MRL/lpr mice. Although not statistically significant, Fli-1(+/-) MRL/lpr mice that received BM from WT MRL/lpr mice also had lower autoantibodies and improved survival compared to WT MRL/lpr mice that received BM from WT MRL/lpr mice. Our data indicate that expression of Fli-1 in haematopoietic cell lineages has a significant effect on disease development in MRL/lpr mice. PMID- 20015091 TI - Pharmacology of epigenetics in brain disorders. AB - Epigenetics is a rapidly growing field and holds great promise for a range of human diseases, including brain disorders such as Rett syndrome, anxiety and depressive disorders, schizophrenia, Alzheimer disease and Huntington disease. This review is concerned with the pharmacology of epigenetics to treat disorders of the epigenome whether induced developmentally or manifested/acquired later in life. In particular, we will focus on brain disorders and their treatment by drugs that modify the epigenome. While the use of DNA methyl transferase inhibitors and histone deacetylase inhibitors in in vitro and in vivo models have demonstrated improvements in disease-related deficits, clinical trials in humans have been less promising. We will address recent advances in our understanding of the complexity of the epigenome with its many molecular players, and discuss evidence for a compromised epigenome in the context of an ageing or diseased brain. We will also draw on examples of species differences that may exist between humans and model systems, emphasizing the need for more robust pre clinical testing. Finally, we will discuss fundamental issues to be considered in study design when targeting the epigenome. PMID- 20015094 TI - An alpha-galactosylceramide C20:2 N-acyl variant enhances anti-inflammatory and regulatory T cell-independent responses that prevent type 1 diabetes. AB - Protection from type 1 diabetes (T1D), a T helper type 1 (Th1)-mediated disease, is achievable in non-obese diabetic (NOD) mice by treatment with alpha galactosylceramide (alpha-GalCer) glycolipids that stimulate CD1d-restricted invariant natural killer T (iNK T) cells. While we have reported previously that the C20:2 N-acyl variant of alpha-GalCer elicits a Th2-biased cytokine response and protects NOD mice from T1D more effectively than a form of alpha-GalCer that induces mixed Th1 and Th2 responses, it remained to determine whether this protection is accompanied by heightened anti-inflammatory responses. We show that treatment of NOD mice with C20:2 diminished the activation of 'inflammatory' interleukin (IL)-12 producing CD11c(high)CD8+ myeloid dendritic cells (mDCs) and augmented the function of 'tolerogenic' DCs more effectively than treatment with the prototypical iNKT cell activator KRN7000 (alpha-GalCer C26:0) that induces Th1- and Th2-type responses. These findings correlate with a reduced capacity of C20:2 to sustain the early transactivation of T, B and NK cells. They may also explain our observation that C20:2 activated iNK T cells depend less than KRN7000 activated iNK T cells upon regulation by regulatory T cells for cytokine secretion and protection from T1D. The enhanced anti-inflammatory properties of C20:2 relative to KRN7000 suggest that C20:2 should be evaluated further as a drug to induce iNK T cell-mediated protection from T1D in humans. PMID- 20015095 TI - Unique activation status of peripheral blood mononuclear cells at acute phase of Kawasaki disease. AB - Although Kawasaki disease (KD) is characterized by a marked activation of the immune system with elevations of serum proinflammatory cytokines and chemokines at acute phase, the major sources for these chemical mediators remain controversial. We analysed the activation status of peripheral blood mononuclear cells (PBMCs) by flow cytometry, DNA microarray and quantitative reverse transcription-polymerase chain reaction. The proportions of CD69+ cells in both natural killer cells and gammadeltaT cells at acute-phase KD were significantly higher than those at convalescent-phase KD. Microarray analysis revealed that five genes such as NAIP, IPAF, S100A9, FCGR1A and GCA up-regulated in acute-phase KD and the pathways involved in acute phase KD were related closely to the innate immune system. The relative expression levels of damage-associated molecular pattern molecule (DAMP) (S100A9 and S100A12) genes in PBMCs at acute-phase KD were significantly higher than those at convalescent-phase KD, while those of TNFA, IL1B and IL6 genes were not significantly different between KD patients and healthy controls. Intracellular production of tumour necrosis factor-alpha, interleukin-10 and interferon-gamma in PBMCs was not observed in KD patients. The present data have indicated that PBMCs showed a unique activation status with high expression of DAMP genes but low expression of proinflammatory cytokine genes, and that the innate immune system appears to play a role in the pathogenesis and pathophysiology of KD. PMID- 20015096 TI - Feto-maternal haemorrhage does not account for the differences in cord blood volume obtained from Black and Asian versus Caucasoid donors. PMID- 20015097 TI - Dic(17;18)(p11.2;p11.2) is a recurring abnormality in chronic lymphocytic leukaemia associated with aggressive disease. AB - Interphase cytogenetics are commonly used to identify clonal abnormalities in chronic lymphocytic leukemia (CLL) patients but fail to identify recurrent translocations that ultimately can direct more focused molecular characterization. Given the importance of del(17p13.1) in CLL outcome, we performed an extensive review of 1213 patients undergoing metaphase cytogenetics at our institution and identified 16 (1.3%) with a recurrent unbalanced translocation between the p arms of chromosomes 17 and 18 that results in a dicentric chromosome with loss of much of 17p and 18p. The dic(17;18)(p11.2;p11.2) was associated with a complex (three or more unrelated cytogenetic abnormalities) karyotype in 12 patients (75%) at the time that the abnormality was first identified, and eventually associated with a complex karyotype in 94% of patients. IGHV mutational analysis was un-mutated in 88% of cases where evaluation was possible. Except for one patient who was diagnosed with CLL incidentally during a workup for metastatic tonsillar cancer, all patients identified with dic(17;18)(p11.2;p11.2) met criteria for disease treatment, with a median time from diagnosis to first treatment of 15 months. Our data demonstrate that dic(17;18)(p11.2;p11.2) is a novel recurrent cytogenetic abnormality in CLL associated with early age at diagnosis and accelerated disease progression. Future efforts to identify genes disrupted by this translocation are warranted and ongoing. PMID- 20015098 TI - Comparing intravitreal triamcinolone acetonide and bevacizumab injections for the treatment of diabetic macular oedema: a randomized double-blind study. AB - PURPOSE: To compare the effect of a single intravitreal injection of triamcinolone acetonide and bevacizumab in reducing macular thickness, which was measured by optical coherence tomography (OCT) in patients with diabetic macular oedema (DMO). METHODS: The patients received a single intravitreal injection of 1.25 mg bevacizumab in one randomly selected eye and 4.0 mg triamcinolone acetonide in the contralateral eye. Central foveal thickness measurement (CFT) with OCT was taken at the initial visit and at the 4-week, 12-week and 24-week visits. RESULTS: Eleven patients (22 eyes) were enrolled and statistically analysed. CFT reduced in the eyes treated with triamcinolone and those treated with bevacizumab in weeks 4 and 12 (p < 0.05). At the 24-week follow-up, no significant difference was noted, relative to the initial visit. Comparing the two groups treated with different drugs, a statistically significant difference in CFT in weeks 4 and 12 was noted, with a more significant reduction in triamcinolone-treated eyes (p < 0.05). Regarding visual acuity (VA), patients treated with triamcinolone had improvement in VA at 4-week (p = 0.02) and 12-week follow-up (p = 0.01), while the group treated with bevacizumab had VA improvement at 4 -week follow-up (p = 0.02). Among the eyes treated with triamcinolone, intraocular pressure (IOP) measurement of more than 21 mmHg was found in three eyes (27.3%). CONCLUSIONS: Intravitreal triamcinolone proved to be more efficient in reducing DMO, providing longer lasting visual improvement, relative to bevacizumab. Eyes treated with triamcinolone had the highest percentage increase in IOP. Further studies are needed to corroborate these findings. PMID- 20015099 TI - Cataract and glaucoma in Korean children with chronic glomerulonephritis receiving systemic corticosteroid treatment. PMID- 20015100 TI - The impact of light source on discrimination ability in subjects with age-related macular degeneration. AB - PURPOSE: To examine the influence of light source on letter contrast sensitivity in subjects with age-related macular degeneration (AMD). METHODS: Halogen incandescent bulbs and low-energy fluorescent tubes were tested with 70 subjects with AMD. The subjects' contrast sensitivity was determined in a randomized single-blind crossover study for each light source using photopically illuminated Pelli Robson contrast sensitivity charts. The test subjects' subjective light source preference was also determined. RESULTS: The mean contrast sensitivity for the incandescent light source was 1.28 +/- 0.29 (mean +/- SD), and for the fluorescent light source 1.17 +/- 0.29, p < 0.001. The illuminance was 338 lux (+/- 9) for the incandescent light, and 339 lux (+/- 11) for the fluorescent light. Forty-nine subjects preferred the incandescent light source, while none preferred the fluorescent light source for maximum detail and clarity. Nineteen had no preference. This finding is statistically significant. Fifteen of the 19 subjects without a preference had no difference in contrast sensitivity, which supports their lack of preference. There was no significant difference with regard to sex or order of exposure to light source. Subjects with AMD had significantly reduced contrast sensitivity compared with expected normal values. We found no relationship between visual acuity and contrast sensitivity. CONCLUSION: We are only able to recommend photopic full spectral radiance incandescent light sources to visually impaired subjects for their domestic surroundings. Furthermore, we recommend the use of full spectral radiance light sources for the illumination of Pelli-Robson contrast sensitivity charts. Given equal illuminance, as in our study, the findings show that contrast sensitivity was better by illumination with incandescent light with full spectral radiance compared with fluorescent light with interrupted spectral radiance. PMID- 20015101 TI - Associations between high myopia and class II HLA. PMID- 20015102 TI - Low-dose azathioprine or mercaptopurine in combination with allopurinol can bypass many adverse drug reactions in patients with inflammatory bowel disease. AB - BACKGROUND: The thiopurine drugs, azathioprine and mercaptopurine (MP), are established treatments for IBD. However, therapeutic failure caused by adverse drug reactions occurs frequently. AIM: To study combination of allopurinol with reduced-dose thiopurine in an attempt to avoid adverse drug reactions in the treatment of IBD. METHODS: Patients with drug reactions to full-dose thiopurines were recruited for combination therapy in two IBD centres in this retrospective study. Dosing was guided by measuring thiopurine methyltransferase (for UK patients) or thioguanine nucleotides and methyl-6MP (Australian patients). Response was monitored by clinical activity indices. RESULTS: Of 41 patients, 25 had non-hepatic and 16 had hepatitic reactions. Clinical remission was achieved in 32 patients (78%) with a median follow-up of 41 weeks (range 0.5-400). Patients who did not respond to combination therapy tended to fail early with the same adverse reaction. The relative risk of having an adverse reaction with methyl-6MP in the top interquartile range was 2.7 (1.3-28) times that with methyl 6MP in the lower three quartiles (95% confidence interval). CONCLUSION: The combined experience from our centres is the largest reported experience of this combination therapy strategy in IBD, and the first to provide evidence for benefit in thiopurine and allopurinol co-therapy to avoid non-hepatitic adverse drug reactions. PMID- 20015103 TI - The development and validation of a new coeliac disease quality of life survey (CD-QOL). AB - BACKGROUND: Previous studies on coeliac disease (CD)-related quality of life (QOL) have been limited by their use of a 'generic' rather than coeliac disease specific assessment instruments. AIM: To develop and psychometrically validate a new coeliac disease-specific instrument, the CD-QOL. METHODS: Through a series of focus groups, we elicited items from patients that related to the specific nature of their disease and its impact on their basic needs. Through expert review, cognitive debriefing with patients and pilot testing, a scale was developed, refined and administered to 387 patients on a gluten-free diet from both community-based support groups and a tertiary care referral centre. Finally, a formal validation study was conducted to assess the psychometric properties of the CD-QOL. RESULTS: The final CD-QOL has 20 items across four clinically relevant subscales (Limitations, Dysphoria, Health Concerns, and Inadequate Treatment). The CD-QOL has high internal consistency, reliability, and psychometric validation indicates both convergent and discriminate validity. CONCLUSIONS: The CD-QOL is a reliable and valid measure of coeliac disease related QOL. As a new disease-specific instrument, it is likely to be a useful tool for evaluating patients with this disorder. PMID- 20015104 TI - The pharmacodynamics and pharmacokinetics of S-tenatoprazole-Na 30 mg, 60 mg and 90 mg vs. esomeprazole 40 mg in healthy male subjects. AB - BACKGROUND: Racemic tenatoprazole 40 mg/day provides more prolonged acid suppression than esomeprazole 40 mg/day. AIM: To compare pharmacodynamic and pharmacokinetic profiles of tenatoprazole and esomeprazole. METHODS: A single centre, double-blind, double-dummy, randomized, 4-way, cross-over study was conducted in 32 healthy male subjects. S-tenatoprazole-Na 30, 60 or 90 mg, or esomeprazole 40 mg was administered once daily for 5 days with 10-day washout intervals. The 24-h intragastric pH was recorded at baseline and on day 5 of each period. RESULTS: On day 5, median pH (5.34 +/- 0.45 and 5.19 +/- 0.52 vs. 4.76 +/ 0.82, respectively, P < 0.002) and percentage time with pH > 4 (80 +/- 11 and 77 +/- 12, vs. 63 +/- 11 respectively, P < 0.0001) for 24-h were higher with S tenatoprazole-Na 90 mg and 60 mg than esomeprazole. In nocturnal periods, S tenatoprazole-Na 90 mg, 60 mg and 30 mg were superior to esomeprazole with regard to median pH (5.14 +/- 0.64, 4.94 +/- 0.65, 4.65 +/- 0.86 and 3.69 +/- 1.18 respectively, P < 0.0001) and percentage time with pH > 4 (77 +/- 12, 73 +/- 17, 64 +/- 17 and 46 +/- 17 respectively, P < 0.0001). Proportion of subjects with nocturnal acid breakthrough with S-tenatoprazole-Na 90 mg, 60 mg and 30 mg was significantly less than with esomeprazole (54.8, 43.3, 56.7 and 90.3 respectively, P < 0.04). The proportion of subjects with >16 hrs with pH >4 was significantly higher with S-tenatoprazole-Na 90 mg and 60 mg than with esomeprazole (87.1%, 83.3% and 41.9% respectively, P < 0.02). CONCLUSIONS: S tenatoprazole-Na produced significantly greater and more prolonged dose-dependent 24-h and nocturnal acid suppression than esomeprazole. S-tenatoprazole-Na may provide greater clinical efficacy compared with current PPIs for patients with ineffective once-daily therapy. PMID- 20015105 TI - The effect of rigid cervical collars on internal jugular vein dimensions. AB - OBJECTIVES: Prior research has demonstrated that rigid cervical collars cause an increase in intracranial pressure (ICP). The mechanism for this effect is unclear and one proposed mechanism involves obstruction of venous outflow in the neck. Ultrasound (US) allows assessment of internal jugular vein dimensions and may yield information regarding the mechanism for the increase in ICP seen with rigid collar application. METHODS: Forty-two healthy volunteers underwent US examination of the internal jugular vein before and after cervical collar application. Internal jugular vein cross-sectional areas were compared with and without the cervical collar in place. RESULTS: The cross-sectional area of the internal jugular vein increased significantly (p < 0.0001) after application of the cervical collar. The mean percentage increase in cross-sectional area was 37% (95% confidence interval [CI] = 20% to 53%). CONCLUSIONS: Internal jugular vein cross-sectional area increases after application of a rigid cervical collar. This supports the hypothesis that venous obstruction in the neck may contribute to the increase in ICP seen after rigid collar application. PMID- 20015106 TI - Mobile crisis team intervention to enhance linkage of discharged suicidal emergency department patients to outpatient psychiatric services: a randomized controlled trial. AB - OBJECTIVES: Many suicidal patients treated and released from emergency departments (ED) fail to follow through with subsequent outpatient psychiatric appointments, often presenting back for repeat ED services. Thus, the authors sought to determine whether a mobile crisis team (MCT) intervention would be more effective than standard referral to a hospital-based clinic as a means of establishing near-term clinical contact after ED discharge. This objective was based on the premise that increased attendance at the first outpatient mental health appointment would initiate an ongoing treatment course, with subsequent differential improvements in psychiatric symptoms and functioning for patients successfully linked to care. METHODS: In a rater-blinded, randomized controlled trial, 120 participants who were evaluated for suicidal thoughts, plans, or behaviors, and who were subsequently discharged from an urban ED, were randomized to follow-up either in the community via a MCT or at an outpatient mental health clinic (OPC). Both MCTs and OPCs offered the same structured array of clinical services and referral options. RESULTS: Successful first clinical contact after ED discharge (here described as "linkage" to care) occurred in 39 of 56 (69.6%) participants randomized to the MCT versus 19 of 64 (29.6%) to the OPC (relative risk = 2.35, 95% CI = 1.55-3.56, p < 0.001). However, we detected no significant differences between groups using intention-to-treat analyses in symptom or functional outcome measures, at either 2 weeks or 3 months after enrollment. We also found no significant differences in outcomes between participants who did attend their first prescribed appointment via MCT or OPC versus those who did not. However divided (MCT vs. OPC, present at first appointment vs. no show), groups showed significant improvements but maintained clinically significant levels of dysfunction and continued to rely on ED services at a similar rate in the 6 months after study enrollment. CONCLUSIONS: Community-based mobile outreach was a highly effective method of contacting suicidal patients who were discharged from the ED. However, establishing initial postdischarge contact in the community versus the clinic did not prove more effective at enhancing symptomatic or functional outcomes, nor did successful linkage with outpatient psychiatric care. Overall, participants showed some improvement shortly after ED discharge regardless of outpatient clinical contact, but nonetheless remained significantly symptomatic and at risk for repeated ED presentations. PMID- 20015107 TI - Diagnosing amaurosis fugax with emergency ultrasound. PMID- 20015109 TI - Multiple sclerosis incidence in the Faroe Islands 1986-2007. AB - OBJECTIVE: Epidemiological studies of the isolated Faroese population in 1945 identified a high annual incidence of multiple sclerosis (MS) of 10/100,000. At the time, there was speculation that the disease was brought to the country by British occupation forces resident in the islands from 1940 to 1945. The objective of the current study is to determine the incidence of diagnosis of MS in the Faroe Islands during the period 1986-2007. METHODS: All patients in the Faroe Islands diagnosed with MS from July 1, 1986 to July 1, 2007 are documented in the current longitudinal, prospective study. The diagnosis is based on clinical observation, magnetic resonance imaging scanning, cerebrospinal fluid tests, and visual evoked potential response testing. RESULTS: The incidence of MS during the period 1986-2007 is 4.5/100,000 annually. This is generally of the same order of magnitude as other research findings in Scandinavia and Iceland. The incidence of MS from 1986 to 2007 is about double the incidence in the Faroe Islands for the period from 1940 to 1986, calculated to be 2.7/100,000 annually. CONCLUSION: The observed incidence of MS in the Faroe Islands, where the population is genetically homogeneous and where the diet exposes the population to neuro-toxic contamination, is at the same level as found in other high-risk regions. The former detected epidemics of MS in Faroe Islands seems apparently to have leveled out and could not be recognized in the recent period covered by the present survey. PMID- 20015108 TI - Changing patterns of antiepileptic drug use in pregnant Australian women. AB - OBJECTIVE: To trace the pattern of antiepileptic drug (AED) use in pregnant Australian women annually from 1999 to 2007, and correlate it with the pattern of AED use in the wider community. METHODS: Analysis of data from the Australian Register of AEDs in Pregnancy, related to Australian population data for AED prescriptions. RESULTS: Over the study period, prescribing of carbamazepine, phenytoin and valproate for pregnant women decreased, and prescribing of lamotrigine, topiramate and levetiracetam increased. These changes tended to parallel prescribing trends in the wider community, except for valproate, whose prescribing in the overall community increased as its prescribing, and its dosage prescribed, decreased in pregnancy. Concomitant with this, there was a trend towards fewer births of foetuses with abnormalities. CONCLUSIONS: While otherwise following national AED prescribing trends, Australian prescribers are reducing the use and dose of valproate in pregnant women, likely in recognition of the teratogenic hazards of this drug. PMID- 20015110 TI - Cutaneous infiltrates and peripheral blood immune responses in dogs with immunomodulatory-responsive lymphocytic-plasmacytic pododermatitis. AB - This study characterizes T- and B-lymphocyte responses in the peripheral blood and lesional skin of dogs with immunomodulatory-responsive lymphocytic plasmacytic pododermatitis (ImR-LPP), a term previously proposed to denote a subpopulation of dogs with idiopathic pododermatitis. T-cell (CD3+, CD4+ and CD8+ ) and B-cell (CD21+) counts were significantly increased in both the epidermis and dermis of lesional ImR-LPP skin compared with that in pedal skin from healthy controls. CD3+ , CD4+, CD8+ and CD21+ cells were commonly observed in perivascular sites in the superficial dermis, periadnexally, beneath the dermal epidermal (DE) junction and in the epidermis of lesional ImR-LPP skin. The CD8+ /CD3+ T-cell ratio in peripheral blood was significantly increased in the ImR-LPP group (0.42 versus 0.35 in controls). Serum IgA, IgG and IgM concentrations were all significantly elevated in affected dogs. Lymphocyte stimulation indices in ImR-LPP dogs were comparable with control levels except for a lower response to ionomycin (6.0 versus 11.1). Dogs with ImR-LPP had a higher incidence and mean (semi-quantitative) score for IgA, IgG and IgM deposits in the epidermis, and a significantly increased incidence of dermal IgA+, IgG+ and IgM+ mononuclear inflammatory cells. The results indicate that upregulated T- and B-lymphocyte responses may contribute to the pathogenesis of the skin lesions observed in dogs with ImR-LPP. PMID- 20015111 TI - GGA autoinhibition revisited. AB - The cytosolic adaptors GGA1-3 mediate sorting of transmembrane proteins displaying a C-terminal acidic dileucine motif (DXXLL) in their cytosolic domain. GGA1 and GGA3 contain similar but intrinsic motifs that are believed to serve as autoinhibitory sites activated by the phosphorylation of a serine positioned three residues upstream of the DXXLL motif. In the present study, we have subjected the widely acknowledged concept of GGA1 autoinhibition to a thorough structural and functional examination. We find that (i) the intrinsic motif of GGA1 is inactive, (ii) only C-terminal DXXLL motifs constitute active GGA binding sites, (iii) while aspartates and phosphorylated serines one or two positions upstream of the DXXLL motif increase GGA1 binding, phosphoserines further upstream have little or no influence and (iv) phosphorylation of GGA1 does not affect its conformation or binding to Sortilin and SorLA. Taken together, our findings seem to refute the functional significance of GGA autoinhibition in particular and of intrinsic GGA binding motifs in general. PMID- 20015112 TI - Thiazolidinediones induce Rab7-RILP-MAPK-dependent juxtanuclear lysosome aggregation and reduce tumor cell invasion. AB - Acidic extracellular pH (pHe) has been shown to stimulate peripheral lysosome trafficking, resulting in cathepsin B secretion and tumor invasion. In addition, inhibitors of sodium-proton exchangers (NHE) such as EIPA, cariporide and s3226, as well as the non-specific NHE inhibitor, troglitazone (Tro), blocked these changes. In this paper, we report a differential ability of the thiazolidinedione (TZD) family of compounds to induce a time-dependent retrograde aggregation of lysosomes over the microtubule-organizing center (MTOC) in tumor cells exposed to acidic pHe. This trafficking event depended on microtubules and the MAP-Kinase pathway, but was independent of Rho GTPase activity. Expression of shRNA implicated Rab7 in this process, and subcellular fractionation revealed that levels of Rab7, RILP and Erk1/2 were increased on lysosomes purified from cells treated with Tro. In addition, DN-RILP overexpression studies indicated that this Rab7 effector also played a role in TZD-induced retrograde trafficking. Tro was able to prevent acidic pHe-induced cell invasion. Finally, DU145 prostate tumor cells stably over-expressing WT-RILP, a condition where lysosomes aggregate to the MTOC in the absence of Tro, did not invade in response to acidic pHe, suggesting that the regulation of lysosome trafficking is an inherently important aspect of tumor cell invasion. PMID- 20015113 TI - Divide et impera: the dictum of peroxisomes. AB - Peroxisomes are unique organelles which display properties of autonomous organelles, as they can multiply by fission of pre-existing ones. Peroxisomes, however, can also develop from the endoplasmic reticulum (ER). This process has convincingly been shown in peroxisome-deficient yeast cells, upon reintroduction of the corresponding gene. Whether peroxisomes also are formed from the ER in wild-type cells that contain peroxisomes is still under debate. Also, the existence of vesicular transport pathways between peroxisomes and the ER is still unresolved. Several new proteins and pathways that play a role in peroxisome proliferation have been identified in the last few years. A surprising finding was that proteins well known for their function in mitochondrial fission (Fis1, Dnm1) are responsible for peroxisome fission as well. In this contribution we discuss recent advancements in research on peroxisome proliferation. PMID- 20015114 TI - Trafficking of the phosphoprotein PfCRT to the digestive vacuolar membrane in Plasmodium falciparum. AB - The digestive vacuole plays an important role in the pathophysiology of the human malaria parasite Plasmodium falciparum. It is a terminal degradation organelle involved in the proteolysis of the host erythrocyte's haemoglobin; it is the site of action of several antimalarial drugs and its membrane harbours transporters implicated in drug resistance. How the digestive vacuole recruits residential proteins is largely unknown. Here, we have investigated the mechanism underpinning trafficking of the chloroquine resistance transporter, PfCRT, to the digestive vacuolar membrane. Nested deletion analysis and site-directed mutagenesis identified threonine 416 as a functional residue for sorting PfCRT to its site of residence. Mass spectroscopy demonstrated that threonine 416 can be phosphorylated. Further phosphorylation was detected at serine 411. Our data establish PfCRT as a phosphoprotein and suggest that phosphorylation of threonine 416 is a possible deciding signal for the sorting of PfCRT to the digestive vacuolar membrane. PMID- 20015115 TI - Bilateral diffuse pulmonary infiltration in a heart transplant recipient. AB - Pulmonary complications are not infrequent after heart transplantation. Kaposi sarcoma is a vascular tumor that can involve the skin as well as visceral organs. We describe a case of visceral and cutaneous Kaposi sarcoma that presented with diffuse bilateral pulmonary infiltration and breathlessness 6 month after heart transplantation. Following modulation of the immunosuppressive regimen and addition of chemotherapy, the patient had an excellent response and has had an uneventful 1-year follow-up. PMID- 20015116 TI - Moods and expectancies of female alcohol drinking--an exploratory study. AB - Gaining access to information concerning mood states and expectations of change preceding a typical drinking occasion is important for understanding the trigger factors for drinking, and for alcohol abuse treatment planning. The objective of the present study was twofold: (i) to explore self-reported states of mood and expectancies preceding a typical drinking occasion vs. relations with parents and drinking outcome; and (ii) to investigate if vulnerability factors in terms of personality and health are related to severity of alcohol problems. The population consisted of 50 women attending a Swedish alcohol clinic. Semi structured interviews were conducted. A mixed-methods design was used encompassing qualitative interview-data and quantitative data from questionnaires and medical journals. Nine out of ten patients had a diagnosis of alcohol dependence, and four out of five had parents with dependency problems. As compared to a female norm group, the patients displayed significantly higher anxiety-related traits and irritability. Moods were described by patients as mostly negative and expectancies of change were evenly distributed between reducing, enhancing or flight from feeling. An expectancy of flight when drinking was also related to a positive relation to mother. The findings pointed to the need for differentiating between coping with and expectancies of drinking. Further, a hierarchical cluster analysis resulted in two groups, indicating one group characterized by higher risk values on personality scales and more severe consequences of drinking. The contribution of a treatment design informed through a gender and culture perspective to treatment outcome was discussed. PMID- 20015117 TI - Factorial validity of the Utrecht Work Engagement Scale (UWES) across occupational groups in Norway. AB - The present study investigated the factorial validity of the Utrecht Work Engagement Scale (UWES) among 1266 participants from ten different occupational groups. Confirmatory factor analyses of the total sample, as well as multi-group analyses and analyses of each of the ten occupational groups separately, indicated that a three-dimensional model of both the UWES-17 and the short version, UWES-9, provided a better fit to the data than a one- and two dimensional model. The results of multi-group analyses and analyses of each of the groups separately, indicated that with a few exceptions, the three-factor model of work engagement provided the best fit. Results indicated factorial invariance and the internal consistencies were acceptable. The fit of the UWES-9 was slightly better than the UWES-17. It is concluded that the Norwegian short version may be recommended over the UWES-17. PMID- 20015118 TI - Effect of organic selenium in the diet on sperm quality of boars. AB - The effect of a diet supplemented with organic selenium (Se) on sperm production and quality of boars was investigated. Sixty mature boars from a commercial artificial insemination centre were randomly allocated at Day (D) 0 into Group A and B. Group A received the regular ration supplemented with inorganic Se (0.4 mg/kg feed as Na(2) SeO(3)) whereas Group B was switched to the same diet but with organic Se (0.4 mg/kg fed as Se-yeast). The sperm was investigated during 4 months (D0, D30, D60, D75, D90, D105 and D120). Sperm concentration and motility were objectively measured using a photometer and Computer Assisted Semen Analysis (CASA) respectively. Morphology of the sperm was assessed using eosin-nigrosin staining and the resistance to induction of oxidative stress (production of malonaldehyde, MDA) through thiobarbituric acid reagent species analysis. Additionally, the Se concentration in sperm and blood plasma were measured. Repeated measures analysis of variance (anova) from D60 to 120 (spermatogenesis of approximately 2 months) or anova at D120 (Se concentrations) were used for statistical analysis. The total number of ejaculated sperm was not significantly different between both groups, but boars of Group B had a significantly higher sperm concentration (434.6 vs 514.1 * 10(6) sperm/ml; p < 0.05). Small differences (p < 0.05) were observed between both groups for some CASA parameters, namely straight line velocity (MUm/s) (Group A: 48.3, Group B: 45.1), straightness (%) (Group A: 65.6, Group B: 62.2) and linearity (%) (Group A: 32.2, Group B: 29.3). The sperm of Group B showed more oxidative stress (4.1 vs 4.9 MUmol MDA/l; p < 0.05) compared with those of Group A. No significant differences (p > 0.05) were observed for the other parameters. Under the present study conditions, changing from inorganic Se to organic Se in the diet of boars increased sperm concentration but reduced some motility parameters and resistance to oxidative stress. PMID- 20015119 TI - Misalignments of rest-activity rhythms in inpatients with schizophrenia. AB - AIMS: Rest-activity rhythms of human beings generally synchronize to a 24-h time cue. Very few detailed research studies have examined rest-activity rhythms in patients with schizophrenia. The present study aimed to explore (i) rest-activity rhythms in patients with schizophrenia, and (ii) factors relevant to their rhythm characteristics. METHODS: We selected only inpatients for this research, because the time cue for inpatients was considered more standardized than that of outpatients. Sixteen inpatients with schizophrenia wore an ActiTrac accelerometer based activity monitor (IM Systems Inc., Baltimore, USA) for eight consecutive days to measure their activity. We used a chi(2) periodogram to compute rest activity rhythms from the activity data, whereby the chi(2) value amplitude was regarded as an index of regularity. We conducted non-parametric tests to identify factors relevant to rhythm cycles and patterns. RESULTS: Half of the participants exhibited prolonged rest-activity cycles, and 25% also had irregular rest activity patterns defined by insufficient chi(2) value amplitude, even though they were clearly under a 24-h time cue. Participants with misaligned rest activity rhythms had attended daytime non-medical treatment programs less frequently, and had received more anti-anxiety/hypnotic medications than those with proper rhythms. CONCLUSION: Changes in rest-activity rhythms by optimizing pharmacological and non-pharmacological treatment could improve social adjustment or quality of life in patients with schizophrenia. PMID- 20015120 TI - Mutation screening and assessment of the effect of genetic variations on expression and RNA editing of serotonin receptor 2C in the human brain. AB - AIM: Serotonin receptor 2C (HTR2C) has been postulated as being involved in the etiology or pathophysiology of mental disorders such as bipolar disorder, major depression and schizophrenia. We previously revealed the altered mRNA expression and RNA editing of HTR2C in the postmortem brains of patients with mental disorders. Here we examined the relationship between genetic variations and expression level or RNA editing level of HTR2C in the human brain. METHODS: We performed mutation screening of the HTR2C gene by sequencing all exons, exon intron boundaries, and promoter region in the same cohort used for expression and RNA editing studies (n = 58). Using the detected genetic variations, we examined the relationship between genetic variations and expression or RNA editing level. RESULTS AND CONCLUSION: We did not find novel mutations or single nucleotide polymorphisms that were specific to patients. Genotype and haplotype-based analyses revealed that genetic variations of HTR2C did not account for observed altered expression or RNA editing level of HTR2C in the brain. PMID- 20015121 TI - Functional alterations in protein kinase C beta II expression in melanoma. AB - Protein kinase C (PKC) is a heterogeneous family of serine/threonine protein kinases that have different biological effects in normal and neoplastic melanocytes (MCs). To explore the mechanism behind their differential response to PKC activation, we analyzed the expression profile of all nine PKC isoforms in normal human MCs, HPV16 E6/E7 immortalized MCs, and a panel of melanoma cell lines. We found reduced PKCbeta and increased PKCzeta and PKCiota expression at both the protein and mRNA levels in immortalized MCs and melanoma lines. We focused on PKCbeta as it has been functionally linked to melanin production and oxidative stress response. Re-expression of PKCbeta in melanoma cells inhibited colony formation in soft agar, indicating that PKCbeta loss in melanoma is important for melanoma growth. PKCbetaII, but not PKCbetaI, was localized to the mitochondria, and inhibition of PKCbeta significantly reduced UV-induced reactive oxygen species (ROS) in MCs with high PKCbeta expression. Thus alterations in PKCbeta expression in melanoma contribute to their neoplastic phenotype, possibly by reducing oxidative stress, and may constitute a selective therapeutic target. PMID- 20015122 TI - Endoreplication in Anemia phyllitidis coincides with the development of gametophytes and male sex. AB - Analyses of DNA content using fluorescence microcytophotometry showed that development of Anemia phyllitidis gametophytes coincided with endoreduplication process. The level of this process shown by the number of endopolyploid cells studied at the I-V arbitrarily established cellular gametophyte stages, was 3%, while at the VI-VII and VII* (male stages) were 10.5 and 4%, respectively. This process coincided with decreased mitotic activity of cells and concerned the cells with their profile area between 1100 and 13,000 microm(2). However, the correlation between cell size and its polyploidisation level was detected only for 12% of these cells. Endoreduplication during development of A. phyllitidis gametophytes seems to be connected with the end of cell cycle followed by the exit of cells from the cell cycle and with subsequent switch of proliferation to the postmitotic differentiation and/or to the endocycle. Endoreplication of A. phyllitidis gametophytes is a function of age, size and number of cells as well as type of gametophyte morphogenesis, which probably maintains the functional copies of genes whose number is restricted by elimination of cells from gametophytes by their death. PMID- 20015123 TI - Expression analysis suggests potential roles of microRNAs for phosphate and arbuscular mycorrhizal signaling in Solanum lycopersicum. AB - MicroRNAs (miRNAs) have emerged as a class of gene expression regulators that play crucial roles in many biological processes. Recently, several reports have revealed that micoRNAs participate in regulation of symbiotic interaction between plants and nitrogen-fixing rhizobia bacteria. However, the role of miRNAs in another type of plant-microbe interaction, arbuscular mycorrhizal (AM) symbiosis, has not been documented. We carried out a microarray screen and poly(A)-tailed reverse transcriptase-polymerase chain reaction (RT-PCR) validation for miRNA expression in tomato (Solanum lycopersicum) under varying phosphate (Pi) availability and AM symbiosis conditions. In roots, miRNA158, miRNA862-3p, miRNA319, miRNA394 and miR399 were differentially regulated under three different treatments, Pi sufficient (+P ), Pi deficient (-P) and AM symbiosis (+M ). In leaves, up to 14 miRNAs were up- or down-regulated under either or both of the Pi treatments and AM symbiosis, of which miR158, miR319 and miR399 were responsive to the treatments in both roots and leaves. We detected that miR395, miR779.1, miR840 and miR867 in leaves were specifically responsive to AM symbiosis, which is independent of Pi availability, whereas miR398 in leaves and miR399 in both roots and leaves were Pi starvation induced. Furthermore, miR158 in roots as well as miR837-3p in leaves were responsive to both Pi deprivation and AM colonization. In contrast, miR862-3p in roots was responsive to Pi nutrition, but not to AM symbiosis. Moreover, the group of miRNA consisting miR319 and miR394 in roots and miR158, miR169g*, miR172, miR172b*, miR319, miR771 and miR775 in leaves were up- and down-regulated by Pi starvation, respectively. The data suggest that altered expression of distinct groups of miRNA is an essential component of Pi starvation-induced responses and AM symbiosis. PMID- 20015124 TI - Transition from pediatric to adult diabetes care: smooth or slippery? AB - OBJECTIVES: The purpose of this study is to evaluate the practices of diabetes health care providers concerning the transition from pediatric to adult diabetes care. The information presented here may help increase awareness of the organization of transitional care for young people with diabetes and prevent the loss of follow-up during this vulnerable period in their lives. METHODS: A questionnaire with an explanatory letter was sent to all members (n = 578) of the International Society for Pediatric and Adolescent Diabetes (ISPAD). A follow-up mailing was sent 4 months later. RESULTS: In total, 92 questionnaires (16%) from members representing 36 countries were included in the analysis. In 76% of the centers, youth are seen until the age of 18 yr; 36% of the pediatric centers see adults > 25 yr; 30% report children under the age of 16 receive follow up from adult diabetologists or internists. About half of the programs already have a structured transition process usually targeting youth 16-25 yr of age. The majority of responders propose that preparation for transition starts at least 1 yr prior to leaving the pediatric center. CONCLUSION: Youth with type 1 diabetes often struggle to keep diabetes management a priority and find it challenging to maintain optimal metabolic control. When they graduate from pediatric care, some of these young people opt out of care altogether, only to resurface in the medical system when they develop complications which may have been prevented. Our survey of diabetes health care professionals in 36 countries worldwide shows that the actual transition practices in many places are far from optimal and require improvement. Transitional care should start early and strategies should promote uninterrupted, comprehensive, and accessible adult care. PMID- 20015125 TI - Wolfram syndrome: important implications for pediatricians and pediatric endocrinologists. PMID- 20015126 TI - Entrainment to distinguish orthodromic reciprocating tachycardia from atrioventricular nodal reentry tachycardia in children. AB - BACKGROUND: Studies in adults suggest that after entrainment from the right ventricle, a post-pacing interval (PPI) minus tachycardia cycle length (TCL), when corrected for atrioventricular node delay (cPPI-TCL), is useful to distinguish atrioventricular nodal reentry tachycardia (AVNRT) from orthodromic reciprocating tachycardia (ORT), but this has not been evaluated in children. METHODS: In 100 children undergoing catheter ablation, entrainment of ORT or AVNRT was performed from the right ventricular apex. The atrial-His (AH) interval was measured on the return cycle (post-AH) and during tachycardia just prior to pacing (pre-AH). The cPPI-TCL was calculated as (PPI-TCL) - (post-AH - pre-AH). In the first 50 children, the best cutoff was identified and then validated in the next 50 children. RESULTS: In the first 50 children, cPPI-TCL was longer in AVNRT compared with ORT (122 +/- 19 ms vs 63 +/- 23 ms, P < 0.001). Furthermore, cPPI-TCL exceeded 95 ms in all AVNRT patients, but was less than < 95 ms in 28 of 29 ORT patients. In the next 50 children, a cPPI-TCL < 95 ms was 100% specific for ORT; a cPPI-TCL > 95 ms was 95% specific for AVNRT. There was even greater separation of cPPI-TCL values comparing AVNRT with ORT utilizing a septal accessory pathway. CONCLUSIONS: The cPPI-TCL is a useful technique to distinguish AVNRT from ORT in children. Our data suggest that in children a cPPI-TCL < 95 ms excludes AVNRT, while a value > 95 ms is rarely observed in ORT. This technique is particularly useful to distinguish AVNRT from ORT utilizing a septal accessory pathway. (PACE 2010; 469-474). PMID- 20015127 TI - Local activation time derived from stored EGM is associated with failure of antitachycardia pacing in patients with implantable defibrillator. AB - BACKGROUND: Antitachycardia pacing (ATP) is an effective treatment of ventricular tachycardia (VT). However, persistent failure of ATP in some patients is well recognized. METHODS: A method of deriving the local activation time from stored intracardiac electrograms in implantable cardioverter defibrillators is described. Using a case-control design, the local activation times were compared between patients with persistent unsuccessful ATP with comparable controls with successful ATP. RESULTS: The local activation times during VT in patients with failed ATP were longer at 120-180 ms compared with corresponding control patients with successful ATP (60-80 ms). The local activation time expressed as a proportion of VT cycle length in patients with failed ATP compared with patients with successful ATP were 0.40 +/- 0.08 versus 0.19 +/- 0.08 (P = 0.012). CONCLUSION: A novel method of deriving local activation time is described, and delayed local activation time may explain failure of ATP in terminating VT in some patients. PMID- 20015128 TI - Effects of changes in the L-type calcium current on hysteresis in restitution of action potential duration. AB - BACKGROUND: Electrical restitution and memory of action potential duration (APD) are considered key determinants of stability of activation. Calcium handling also impacts stability of activation via its modification of the above and independently. We determined the effects of reduction in an important calcium current, I(CaL), on hysteresis in restitution of APD, which provides assessment of restitution and memory. METHODS: Transmembrane potentials were recorded from isolated ventricular tissues from pigs. The pacing protocols consisted of sequential and oscillatory changes in diastolic intervals (DI). Two protocols were used, one where DIs ranged between 100 and 700 ms and the other where DIs ranged between 10 and 290 ms. Verapamil (2 microM/L) was used to reduce I(CaL). From the hysteresis observed in restitution, loop area, thickness, overall tilt, and the maximum and minimum delays between DIs and APDs were calculated to obtain measures of memory and restitution. RESULTS: Restitution during sequential and oscillatory changes in DI displayed hysteresis. All measures of hysteresis decreased after reduction in I(CaL). The decreases in area, thickness, and overall tilt were most pronounced (50%, 50%, and 33%). CONCLUSION: Sequential and oscillatory changes in DIs reveal that while the overall slope of restitution of APD decreases with decrease in I(CaL), so do measures of memory. Because slopes and memory are considered to have contrasting effects on stability, our results suggest that predicting effects of alteration in I(CaL) on stability of activation using slope alone may not be adequate. (PACE 2010; 451-459). PMID- 20015129 TI - DDD versus VVIR pacing in patients, ages 70 and over, with complete heart block. AB - BACKGROUND: Dual-chamber pacing is believed to have an advantage over single chamber ventricular pacing. The aim of the study was to determine whether elderly patients with implanted pacemaker for complete atrioventricular block gain significant benefit from dual-chamber (DDD) compared with single-chamber ventricular demand (VVIR). METHODS: The study was designed as a double-blind randomized two-period crossover study-each pacing mode was maintained for 3 months. Thirty patients (eight men, mean age 76.5 +/- 4.3 years) with implanted PM were submitted to a standard protocol, which included an interview, functional class assessment, quality of life (QoL) questionnaires, 6-minute walk test, and transthoracic echocardiographic examinations. QoL was measured by the SF-36. All these parameters were obtained on DDD mode pacing and VVIR mode pacing. Paired data were compared. RESULTS: QoL was significantly different between the two groups and showed the best values in DDD. Overall, no patient preferred VVIR mode, 18 preferred DDD mode, and 12 expressed no preference. No differences in mean walking distances were observed between patients with single-chamber and dual-chamber pacing. VVI pacing elicited marked decrease in left ventricle ejection fraction and significant enlargement of the left atrium. DDD pacing resulted in significant increase of the peak systolic velocities in lateral mitral annulus and septal mitral annulus. Early diastolic velocities on both sides of mitral annulus did not change. CONCLUSION: In active elderly patients with complete heart block, DDD pacing is associated with improved quality of life and systolic ventricular function compared with VVI pacing. PMID- 20015130 TI - Inappropriate biventricular implantable cardioverter defibrillator firing due to cryptogenic double counting. PMID- 20015132 TI - Unusual ECG pattern of right atrial appendage atrial tachycardia in one patient with right pneumonectomy. AB - The right atrial appendage atrial tachycardia (RAA AT) has been previously reported as a rare site in focal AT. We report a patient with a history of a right pneumonectomy who underwent catheter ablation of the AT originating from the RAA. This RAA AT showed unusual P-wave morphology compared with previous reports. We describe the RAA AT following right pneumonectomy using a NavX system (St. Jude Medical, St. Paul, MN, USA). PMID- 20015131 TI - Postural orthostatic tachycardia syndrome: a rare complication following electrical injury. AB - We report on two previously healthy patients who developed severe form of postural orthostatic tachycardia syndrome (POTS) following an electric injury. Both the patients developed symptoms of orthostatic intolerance in the form of dizziness, fatigue, lightheadedness, and palpitations, weeks to months after electrical injury. Orthostatic intolerance produced considerable functional impairment in these patients. Early recognition of POTS when it occurs after an electrical injury allows for prompt evaluation and management to occur. PMID- 20015133 TI - Successful catheter ablation of reentrant junctional tachycardia in a patient with asplenia syndrome before total cavo-pulmonary connection. AB - Asplenia syndrome is commonly associated with complex structural cardiac malformations, and junctional tachycardia (JT), which may compromise hemodynamic status, has been reported in association with asplenia syndrome.(1) We report successful radiofrequency catheter ablation of reentrant JT in a patient with asplenia syndrome. PMID- 20015134 TI - Electrical characteristics of an electronic control device under a physiologic load: a brief report. AB - BACKGROUND: Law enforcement officers use electronic control devices (ECDs), such as the TASER X26 (TASER International, Inc., Scottsdale, AZ, USA), to control resisting subjects. Some of the debate on the safety of the devices has centered on the electrical characteristics of the devices. The electrical characteristics published by TASER International have historically based on discharges into a 400 Omega resistor. There are no studies that the authors are aware of that report the electrical characteristics under a physiologic load. In this study, we make an initial attempt to determine the electrical characteristics of the TASER X26 during a 5-second exposure in human volunteers. METHODS: Subjects received an exposure to the dry, bare chest (top probe), and abdomen (bottom probe) with a standard TASER X26 in the probe deployment mode for 5 seconds. There were 10-11 pulse captures during the 5 seconds. Resistance was calculated using the sum of the absolute values of the instantaneous voltage measurements divided by the sum of the absolute values of the instantaneous current measurements (Ohm's Law). RESULTS: For the eight subjects, the mean spread between top and bottom probes was 12.1 inches (30.7 cm). The mean resistance was 602.3 Omega, with a range of 470.5-691.4 Omega. The resistance decreased slightly over the 5-second discharge with a mean decrease of 8.0%. The mean rectified charge per pulse was 123.0 microC. The mean main phase charge per pulse was 110.5 microC. The mean pulse width was 126.9 micros. The mean voltage per pulse was 580.1 V. The mean current per pulse was 0.97 A. The average peak main phase voltage was 1899.2 V and the average peak main phase current was 3.10 A. CONCLUSIONS: The mean tissue resistance was 602.3 Omega in this study. There was a decrease in resistance of 8% over the 5-second exposure. This physiologic load is different than the 400 Omega laboratory load used historically by the manufacturer. We recommend future characterization of these devices use a physiologic load for reporting electrical characteristics. We also recommend that all the electrical characteristics be reported. PMID- 20015135 TI - Differential manifestation of cardiovascular complaints as a function of utilization of ergogenic supplements. AB - INTRODUCTION: The rate of use of dietary supplements among young adults is significant. While the military makes significant restrictions on the use of certain pharmacologic drugs and actively tests for illegal drugs in a deployed environment, there is a near-unlimited supply of body-enhancing supplements available at military exchanges to deployed personnel. By emphasizing physical performance and providing these for purchase, the military leadership, perhaps unknowingly, endorses the use of these products. Cardiovascular symptoms represent one of the leading nontraumatic causes of aeromedical evacuation from a combat zone. Whether the use of supplements is associated with a differential presentation to cardiovascular complaint is unknown. METHODS: Retrospective review using the US Department of Defense Military Health System data, we identified patients evaluated for cardiovascular complaints of syncope or palpitations while deployed to Iraq and Afghanistan. RESULTS: There were 905 US military personnel who presented with complaint of syncope or palpitations (mean age 31 +/- 10 years, 77% male). There were 83 (9.2%) who self-reported taking an ergogenic supplement. The incidence of reported use of supplements among males was 10.8%, which was significantly higher than its use among females at 3.8% (P = 0.001). In those >30 years, those on supplements had a higher resting pulse (90 +/- 28 vs 79 +/- 24 beats/min, P = 0.032), and the incidence of resting tachycardia was three-fold higher (35.0% vs 11.4%, P = 0.008). Supplement use was seen in 12.3% of those who presented with palpitations, which was significantly higher than those who presented without palpitations (7.8%, P = 0.043). In those taking supplements, symptoms were more likely during exertion (26.5% vs 15.0%, P < 0.001), and immediately postexertional (13.2% vs 4.6%, P < 0.001). An electrocardiogram was suggestive of diagnosis in 103 (16.3%), while head computed tomography, treadmill, and echocardiogram had no diagnostic utility in this patient population. DISCUSSION: In a healthy population serving within a combat zone, there exists a differential expression of disease in those taking supplements. Further study of a prospective nature to determine the impact of supplement use in this environment may allow for a more refined policy toward use and medical evaluation. PMID- 20015136 TI - Termination of electrocution-induced ventricular fibrillation by an implantable cardioverter defibrillator. AB - We describe a case in which an implantable cardioverter defibrillator (ICD) saved a patient's life after he tried to commit suicide by electrocuting himself. Deaths caused by electrocution are usually accidental and rarely suicidal. The amount of current flow is the most important factor in deciding the degree of electrical injury, which may range from transient muscle tremors to death. The ICD is electrically insulated from surrounding electromagnetic interference and the passage of electric current typically does not damage or reprogram the device. In our patient, electromagnetic interference caused by the electric current initially triggered the noise reversion mode, leading to asynchronous pacing. Ventricular fibrillation was detected and terminated only after the electromagnetic interference stopped, as depicted by the intracardiac electrogram. This case is the first documented example of an ICD-aborted fatal electrocution from ventricular fibrillation caused by an unnatural electrical source. (PACE 2010; 510-512). PMID- 20015137 TI - Sedation trends in the 21st century: the transition to dexmedetomidine for radiological imaging studies. AB - Sedation for radiological imaging studies encompasses the majority of all sedation-related procedures outside of the intensive care unit. This review will follow the evolution of pediatric sedation for radiological imaging studies in North America as well as the transition of sedation services from the oversight of radiologists to those of other providers. The evolving options for sedation agents will be reviewed, with attention given to examining the advantages, limitations, and risks of replacing the standard sedatives with dexmedetomidine. PMID- 20015138 TI - Dexmedetomidine: perioperative applications in children. AB - Dexmedetomidine is a highly specific and selective alpha-2-adrenergic agonist with sedative, anxiolytic, and organ protective effects. Its clinical applications in children include premedication, prevention of emergence delirium, as part of multimodal anesthetic regimen and sedation in the pediatric intensive care unit. Its role in neuroprotection in children undergoing anesthesia should be explored. In this review, various uses of dexmedetomidine are discussed in detail. PMID- 20015139 TI - Spinal needle design and size affect the incidence of postdural puncture headache in children. AB - BACKGROUND: In adults, pencil point spinal needles are known to be less traumatic and hence to be superior compared with cutting point needles in respect of postpuncture complications. In children, only a few trials have evaluated the difference in the incidence of postdural puncture headache (PDPH) using spinal needles with different tip designs. The aim of this study was to evaluate the success rate and the incidence of PDPH and backache following spinal anesthesia (SA) with the two types of needles currently in use for children. METHODS: This is a retrospective study of prospectively collected data. The success rate and postpuncture complications of 26G cutting point (Atraucan) spinal needle were compared with 27G pencil point (Pencan) spinal needle in 414 children aged 2-17 years undergoing surgery with SA. RESULTS: Both needles had similar first-attempt success rates: 87% in the cutting point group and 91% in the pencil point group (P = 0.16). Pencil point needles caused less PDPH compared to cutting point needles; 0.4% vs 4.5%, respectively (P = 0.005). Both needles caused similar backache (P = 0.08). No severe neurologic symptom was reported for both needles. CONCLUSION: The data suggest that 27G pencil point spinal needles lead to less PDPH compared to 26G cutting point spinal needles in children. PMID- 20015140 TI - Microgeographic population structure of green swordail fish: genetic differentiation despite abundant migration. AB - Swordtails (Xiphophorus; Poeciliidae) have figured prominently in research on fish mating behaviours, sexual selection, and carcinogenesis, but their population structures and dispersal patterns have been relatively neglected. Using nine microsatellite loci, we estimated genetic differentiation in Xiphophorus helleri within and between adjacent streams in Belize. The genetic data were complemented by a tagging study of movement within one stream. In the absence of physical dispersal barriers (waterfalls), population structure followed an isolation by distance (IBD) pattern. Genetic differentiation (F(ST) up to 0.07) was significant between and within creeks, despite high dispersal in the latter as judged by the tagging data. Such heterogeneity apparently was a result of genetic drift in local demes, due to small population sizes and highly skewed paternity. The IBD pattern was interrupted by waterfalls, boosting F(ST) above 0.30 between adjacent samples across these barriers. Overall, our results are helpful in understanding the interplay of evolutionary forces and population dynamics in a small fish living in a changeable habitat. PMID- 20015141 TI - Changes in genomic methylation patterns during the formation of triploid asexual dandelion lineages. AB - DNA methylation is an epigenetic mechanism that has the potential to affect plant phenotypes and that is responsive to environmental and genomic stresses such as hybridization and polyploidization. We explored de novo methylation variation that arises during the formation of triploid asexual dandelions from diploid sexual mother plants using methylation-sensitive amplified fragment length polymorphism (MS-AFLP) analysis. In dandelions, triploid apomictic asexuals are produced from diploid sexual mothers that are fertilized by polyploid pollen donors. We asked whether the ploidy level change that accompanies the formation of new asexual lineages triggers methylation changes that contribute to heritable epigenetic variation within novel asexual lineages. Comparison of MS-AFLP and AFLP fragment inheritance in a diploid x triploid cross revealed de novo methylation variation between triploid F(1) individuals. Genetically identical offspring of asexual F(1) plants showed modest levels of methylation variation, comparable to background levels as observed among sibs in a long-established asexual lineage. Thus, the cross between ploidy levels triggered de novo methylation variation between asexual lineages, whereas it did not seem to contribute directly to variation within new asexual lineages. The observed background level of methylation variation suggests that considerable autonomous methylation variation could build up within asexual lineages under natural conditions. PMID- 20015142 TI - Deep phylogeographic divisions and long-term persistence of forest invertebrates (Hexapoda: Collembola) in the North-Western Mediterranean basin. AB - The North-Western Mediterranean basin is well known for its high number of relictual endemic taxa, and has been indicated as one of the world's major biodiversity hotspots at the species level. A possible contributing factor may be long-term persistence of populations and their prolonged stability. This study was designed to investigate the phylogeographic structure of three common species of the genus Lepidocyrtus (Hexapoda: Collembola), soil-dwelling arthropods characterized by limited dispersal capabilities and generally associated with forest habitats. We observed a remarkable geographic structure, with numerous deeply divergent genetic lineages occupying islands as well as mainland sites with no apparent gene flow among most sites, even across distances of only tens of kilometres. The reconstructed time frame for the evolution of these lineages suggests divergence between 5 and 15 Ma. This indicates a remarkably ancient origin and long-term persistence of individual lineages over a fine geographic scale despite the occurrence of abrupt sea level and climatic fluctuations in the area. This further suggests that currently recognized morphological species might be a serious underestimation of the true springtail biodiversity within this region. PMID- 20015143 TI - Genome scan in the mosquito Aedes rusticus: population structure and detection of positive selection after insecticide treatment. AB - Identification of genes involved in local adaptation is particularly challenging for species functioning as a network of interconnected populations undergoing frequent extinctions-recolonizations, because populations are submitted to contrasted evolutionary pressures. Using amplified fragment length polymorphism markers, population genetic structure of the mosquito Aedes rusticus was analysed in five geographical areas of the French Rhone-Alpes region. We included a number of sites that were treated with the bio-insecticide Bacillus thuringiensis israelensis (Bti) for more than 15 years. Analysis of molecular variance revealed that most of the genetic variability was found within populations (96%), with no significant variation among geographical areas, although variation among populations within areas (4%) was significant. The global genetic differentiation index F(ST) was low (0.0366 +/- 0.167). However, pairwise F(ST) values were significant and no isolation-by-distance at the regional level was observed, suggesting a metapopulation structure in this species. Bti-treatment had no effect on genetic structure and on within-population genetic diversity. Potential signatures of positive selection associated with Bti-treatment were detected for five loci, even though toxicological bioassays performed on field-collected larvae showed no significant difference in mortality between Bti-treated and nontreated sites. The difficulty of detecting moderate resistance in field collected larvae together with possible differential persistence of toxins in the environment may explain our inability to detect a toxicological response to Bti in treated sites. The evidence for positive selection occurring at several genomic regions suggests a first step towards Bti resistance in natural mosquito populations treated with this bio-insecticide. Furthermore, this signal was detectable using genomic tools before any toxicological evidence for resistance could be identified. PMID- 20015144 TI - Molecular identification of brood-parasitic females reveals an opportunistic reproductive tactic in ruddy ducks. AB - In many taxa, females lay eggs in the nests of other conspecifics. To determine the conditions under which conspecific brood parasitism develops, it is necessary to identify parasitic offspring and the females who produce them; however, for most systems parasitism can be difficult to observe and most genetic approaches have relatively low resolving power. In this study, we used protein fingerprinting from egg albumen and 10 microsatellite loci to genetically match parasitic ducklings to their mothers in a population of ruddy ducks (Oxyura jamaicensis). We found that 67% of nests contained parasitic offspring, and we successfully identified their mothers in 61% of the cases. Of the parasitic females identified, 77% also had nests of their own (i.e. a dual tactic, where females both nest and lay parasitically), and we found no evidence that parasitic females pursued a specialist (parasitism only) tactic. We also found that parasitic egg laying was not influenced by nest loss, predation or female condition. Thus, in contrast to most waterfowl studied to date, female ruddy ducks appear to lay parasitic eggs whenever the opportunity arises. PMID- 20015145 TI - A multi-protein complex controls cAMP signalling and filamentation in the fungal pathogen Candida albicans. AB - Candida albicans is an opportunistic fungal pathogen of humans. The ability of the fungus to grow as both yeast and filamentous forms is essential for its pathogenicity. Morphogenesis of C. albicans is largely regulated through the secondary messenger cAMP, produced by the soluble adenylyl cyclase, Cyr1p. Recent evidence suggests that Cyr1p can be directly stimulated by environmental cues to increase cytoplasmic cAMP levels and thus promote hyphal development. In this issue of Molecular Microbiology, Zou et al. demonstrate that, in response to some environmental cues, Cyr1p functions as part of a tripartite complex additionally involving Cap1p and G-actin. All three proteins in the complex are required to raise cytosolic cAMP levels after stimulation with serum and bacterial peptidoglycan. The formation of such a complex highlights the importance of precise regulation of Cyr1p activity in response to host environmental cues. PMID- 20015146 TI - Horizontally acquired homologues of the nucleoid-associated protein H-NS: implications for gene regulation. AB - H-NS is one of the most intensively studied members of the family of bacterial nucleoid-associated proteins. It is a DNA-binding protein with a preference for A+T-rich DNA sequences, and it represses the transcription of hundreds of genes in Gram-negative bacteria, including pathogens. In most cases where the issue has been investigated, the repressive activity of H-NS is opposed by the intervention of an antagonistically acting DNA-binding protein, a remodelling of local DNA structure, or a combination of these two. H-NS activity can also be modulated by protein-protein interaction with members of the Hha/YdgT protein family, molecules that share partial amino acid sequence similarity to the oligomerization domain of H-NS. Of particular interest is the ability of H-NS to interact with the full-length paralogue StpA or full-length orthologues that have been acquired by horizontal DNA transfer. In this issue of Molecular Microbiology, Muller et al. describe the H-NS orthologue Hfp and present evidence that in bacteria that acquire Hfp the range of activities of H-NS is modified with important implications for the physiology of the bacterium. PMID- 20015147 TI - Induction of the ferritin gene (ftnA) of Escherichia coli by Fe(2+)-Fur is mediated by reversal of H-NS silencing and is RyhB independent. AB - FtnA is the major iron-storage protein of Escherichia coli accounting for < or = 50% of total cellular iron. The FtnA gene (ftnA) is induced by iron in an Fe(2+) Fur-dependent fashion. This effect is reportedly mediated by RyhB, the Fe(2+)-Fur repressed, small, regulatory RNA. However, results presented here show that ftnA iron induction is independent of RyhB and instead involves direct interaction of Fe(2+)-Fur with an 'extended' Fur binding site (containing five tandem Fur boxes) located upstream (-83) of the ftnA promoter. In addition, H-NS acts as a direct repressor of ftnA transcription by binding at multiple sites (I-VI) within, and upstream of, the ftnA promoter. Fur directly competes with H-NS binding at upstream sites (II-IV) and consequently displaces H-NS from the ftnA promoter (sites V-VI) which in turn leads to derepression of ftnA transcription. It is proposed that H-NS binding within the ftnA promoter is facilitated by H-NS occupation of the upstream sites through H-NS oligomerization-induced DNA looping. Consequently, Fur displacement of H-NS from the upstream sites prevents cooperative H-NS binding at the downstream sites within the promoter, thus allowing access to RNA polymerase. This direct activation of ftnA transcription by Fe(2+)-Fur through H-NS antisilencing represents a new mechanism for iron induced gene expression. PMID- 20015148 TI - Acute liver injury upregulates microRNA-491-5p in mice, and its overexpression sensitizes Hep G2 cells for tumour necrosis factor-alpha-induced apoptosis. AB - BACKGROUND: MicroRNAs (miRNAs) have emerged as novel genetic regulators of cell functions such as proliferation, apoptosis and cancer. AIMS: The aim of this study was to evaluate the role of a specific miRNA in modulating hepatic cell functions. METHODS: C57Bl/6 mice were administered anti-fas receptor antibodies to induce liver cell apoptosis. miRNAs were purified from the liver tissue and evaluated using an miRNA microarray. The role of miRNA-491_5p, which was overexpressed in the model, in modulating hepatic cell functions was evaluated. miRNA-491_5p was overexpressed in Hep G2 cells, followed by the addition of tumour necrosis factor (TNF)-alpha, and induction of apoptosis as well as genes involved in apoptosis pathways were evaluated. The effect of miRNA-491_5p target genes on apoptosis was also analysed by inhibiting their expression by siRNA induced gene silencing. RESULTS: Upregulation of miRNA-491_5p was found in a high dose anti-fas receptor antibody group. Overexpression of microRNA-491_5p sensitized Hep G2 cells for TNF-alpha-induced apoptosis, and also caused an inhibition of alpha-fetoprotein, (AFP), heat shock protein-90 (hsp-90) and nuclear factor-kappaB (NF-kappaB). Overexpression of miRNA-491_5p or inhibition of AFP and hsp-90 resulted in an increased apoptosis in TNF-alpha-treated Hep G2 cells. CONCLUSIONS: One of the miRNAs that is associated with the acute liver injury mouse model, miRNA-491_5p, sensitizes Hep G2 cells for TNF-alpha-induced apoptosis, at least in part, by inhibiting AFP, hsp-90 and NF-kappaB. PMID- 20015149 TI - The neglected hepatitis C virus genotypes 4, 5 and 6: an international consensus report. AB - Abstract Hepatitis C virus (HCV) genotypes 4, 5 and 6 represent >20% of all HCV cases worldwide. HCV-4 is mainly seen in Egypt, where it represents 90% of all HCV cases. Antischistosomal therapy was the main cause of contamination there, followed by procedures performed by informal providers and traditional healers such as dental care, wound treatment, circumcision, deliveries, excision and scarification. It is also highly prevalent in sub-Saharan Africa and in the Middle East. In Europe, its prevalence has recently increased particularly among intravenous drug users and in immigrants. HCV-5 is mainly found in South Africa, where it represents 40% of all HCV genotypes, but four pockets of HCV-5 were found in France, Spain, Syria and Belgium and sporadic cases were found elsewhere. The mode of transmission is mainly iatrogenic and transfusion. HCV-6 is found in Hong Kong, Vietnam, Thailand and Myanmar and also in American and Australian from Asian origin. The response to treatment in HCV-4 is intermediate between HCV-1 and HCV-2 and HCV-3. A sustained viral response is achieved in 43 70% with pegylated interferon and ribavirin. It is higher in Egyptians than Europeans and Africans and is negatively related to insulin resistance and to the severity of fibrosis. It increases to >80% with 24 weeks of therapy only if a rapid virological response is achieved. In HCV-5, a sustained virological response is achieved in >60% with 48 weeks of therapy. HCV-6 is also considered an easy-to-treat genotype, leading to a response in 60-85% of cases. PMID- 20015150 TI - An unusual cause of jaundice in pregnancy. PMID- 20015151 TI - Acute phase proteins and bacterial infections in cirrhosis. PMID- 20015152 TI - A levitating tattoo in a hepatitis C patient on treatment. PMID- 20015153 TI - A novel choline cotransporter sequestration compartment in cholinergic neurons revealed by selective endosomal ablation. AB - The sodium-dependent, high affinity choline transporter - choline cotransporter - (ChCoT, aka: cho-1, CHT1, CHT) undergoes constitutive and regulated trafficking between the plasma membrane and cytoplasmic compartments. The pathways and regulatory mechanisms of this trafficking are not well understood. We report herein studies involving selective endosomal ablation to further our understanding of the trafficking of the ChCoT. Selective ablation of early sorting and recycling endosomes resulted in a decrease of approximately 75% of [3H]choline uptake and approximately 70% of [3H]hemicholinium-3 binding. Western blot analysis showed that ablation produced a similar decrease in ChCoTs in the plasma membrane subcellular fraction. The time frame for this loss was approximately 2 h which has been shown to be the constitutive cycling time for ChCoTs in this tissue. Ablation appears to be dependent on the intracellular cycling of transferrin-conjugated horseradish peroxidase and the selective deposition of transferrin-conjugated horseradish peroxidase in early endosomes, both sorting and recycling. Ablated brain slices retained their capacity to recruit via regulated trafficking ChCoTs to the plasma membrane. This recruitment of ChCoTs suggests that the recruitable compartment is distinct from the early endosomes. It will be necessary to do further studies to identify the novel sequestration compartment supportive of the ChCoT regulated trafficking. PMID- 20015155 TI - Macromolecular drug transport into the brain using targeted therapy. AB - The brain forms a vascular barrier system comprised of the blood-brain barrier (BBB) and the blood-CSF barriers. Together they prevent the passage of a number of drugs from the bloodstream into the brain parenchyma, because their molecules are either hydrophilic, too large or both. In many disorders affecting the CNS, these barriers are physically intact, which limits the entry of large molecules with potentially important therapeutic implications. The BBB is the most relevant barrier against drug delivery to the brain as the area of the BBB is about 1000 times larger than that of the blood-CSF barrier. Moreover, the transport through the choroid plexus is directed to the ventricular system, only allowing the transported molecules to access cells near the ventricular and subarachnoid surfaces. This review outlines possible routes for targeted entry of macromolecules like polypeptides, siRNA and cDNA. In the vascular compartment, targeting molecules should interact specifically with proteins expressed exclusively by these barrier cells, and therefore prevent uptake elsewhere in the body. Preferably, the targeting molecule should be conjugated to a drug carrier that allows uptake of a defined cargo. However, evidence for transport of such targetable drug-carrier complexes through the barriers, in particular the BBB, is contentious, and is discussed with emphasis on the different attempts that have evinced transport through the BBB not only from blood-to-endothelium, but also from endothelium-to-brain. PMID- 20015154 TI - Cholesterol modulates ion channels via down-regulation of phosphatidylinositol 4,5-bisphosphate. AB - Ubiquitously expressed Mg(2+)-inhibitory cation (MIC) channels are permeable to Ca2+ and Mg2+ and are essential for cell viability. When membrane cholesterol level was increased by pre-incubating cells with a water-soluble form of cholesterol, the endogenous MIC current in HEK293 cells was negatively regulated. The application of phosphatidylinositol 4,5-bisphosphate (PIP2) recovered MIC current from cholesterol effect. As PIP2 is the direct modulator for MIC channels, high cholesterol content may cause down-regulation of PIP2. To test this possibility, we examined the effect of cholesterol on two exogenously expressed PIP2-sensitive K+ channels: human Ether-a-go-go related gene (HERG) and KCNQ. Enrichment with cholesterol inhibited HERG currents, while inclusion of PIP2 in the pipette solution blocked the cholesterol effect. KCNQ channel was also inhibited by cholesterol. The effects of cholesterol on these channels were blocked by pre-incubating cells with inhibitors for phospholipase C, which may indicate that cholesterol enrichment induces the depletion of PIP2 via phospholipase C activation. Lipid analysis showed that cholesterol enrichment reduced gamma-(32)P incorporation into PIP2 by approximately 35%. Our results suggest that cholesterol may modulate ion channels by changing the levels of PIP2. Thus, an important cross-talk exists among two plasma membrane-enriched lipids, cholesterol and PIP2. PMID- 20015156 TI - Prospective multicenter assessment of interobserver agreement for radiologist interpretation of multidetector computerized tomographic angiography for pulmonary embolism. AB - BACKGROUND: Emergency physicians rely on the interpretation of radiologists to diagnose and exclude pulmonary embolism (PE) on the basis of computerized tomographic pulmonary angiography (CTPA). Few data exist regarding the interobserver reliability of this endpoint. OBJECTIVE: To quantify the degree of agreement in CTPA interpretation between four academic hospitals and an independent reference reading (IRR) laboratory. METHODS: Hospitalized and emergency department patients who had one predefined symptom and sign of PE and underwent 64-slice CTPA were enrolled from four academic hospitals. CTPA results as interpreted by board-certified radiologists from the hospitals were compared against those from the IRR laboratory. CTPAs were read as indeterminate, PE(-) or PE(+), and percentage obstruction was computed by the IRR laboratory, using a published method. Agreement was calculated with weighted Cohen's kappa. RESULTS: We enrolled 492 subjects (63% female, age 54 +/- 1 years, and 16.7% PE(+) at the site hospitals). Overall agreement was 429/492 (87.2%; 95% confidence interval 83.9-90.0). We observed 13 cases (2.6%) of complete discordance, where one reading was PE(+) and the other reading was PE(-). Weighted agreement was 92.3%, with kappa = 0.75. The median percentage obstruction for all patients was 9% (25th-75th percentile interquartile range: 5% to -30%). For CTPAs interpreted at the site hospitals as PE(-) or indeterminate but read as PE(+) by the IRR laboratory, the median of percentage obstruction was 6% (4-7%). CONCLUSION: We found in this sample a good level of agreement, with a weighted kappa of 0.75, but with 2.6% of patients having total discordance. Overall, a large proportion of clots were distal or minimally occlusive clots. PMID- 20015157 TI - 25-hydroxy-vitamin D levels among Callithrix penicillata primate species raised in captivity. AB - BACKGROUND: Animals in captivity should receive adequate sunlight exposure for sufficient generation of vitamin D [25(OH)D]. In the present study, 25(OH)D serum levels of 84 Callithrix penicillata primates were evaluated. OBJECTIVES: To determine 25(OH)D levels of those animals; to evaluate the influence of gender and period of sunlight exposure on their 25(OH)D levels. METHODS: Three groups were evaluated: group 1 (n = 29) on free sunlight exposure; group 2 (n = 34) on partial sunlight exposure; group 3 (n = 21) without sunlight exposure. RESULTS: The obtained 25(OH)D values were: group 1, 121.2 +/- 33.3 ng/ml; group 2, 115.2 +/- 32.2 ng/ml; group 3, 53.3 +/- 10.4 ng/ml. Significant statistical differences were obtained between groups 1 and 3 (p < 0.001) and groups 2 and 3 (p < 0.001); no statistical difference was found between genders. CONCLUSION: Direct sunlight exposure is essential for 25(OH)D sufficiency and it is proposed that the 25(OH)D normal range for captive Callithrix penicillata would be from 104.8 to 137.1 ng/ml (CI = 95%). PMID- 20015158 TI - Magnetic activated cell sorting allows isolation of spermatogonia from adult primate testes and reveals distinct GFRa1-positive subpopulations in men. AB - BACKGROUND: Isolation of spermatogonial stem cells (SSCs) could enable in vitro approaches for exploration of spermatogonial physiology and therapeutic approaches for fertility preservation. SSC isolation from adult testes is difficult due to low cell numbers and lacking cell surface markers. Glial cell derived neurotrophic factor family receptor alpha-1 (GFRalpha1) plays a crucial role for the maintenance of SSCs in rodents and is expressed in monkey spermatogonia. METHODS: Magnetic activated cell sorting was employed for the enrichment of GFRalpha1+ spermatogonia from adult primate testes. RESULTS: Magnetic activated cell sorting of monkey cells enriched GFRalpha1+ cells threefold. 11.4% of GFRalpha1+ cells were recovered. 42.9% of GFRalpha1+ cells were recovered in sorted fractions of human testicular cells, representing a fivefold enrichment. Interestingly, a high degree of morphological heterogeneity among the GFRalpha1+ cells from human testes was observed. CONCLUSIONS: Magnetic activated cell sorting using anti-GFRalpha1 antibodies provides an enrichment strategy for spermatogonia from monkey and human testes. PMID- 20015159 TI - Analysis of humoral immune responses in rhesus macaques vaccinated with attenuated SIVmac239Deltanef and challenged with pathogenic SIVmac251. AB - BACKGROUND: To determine the correlation between protection and humoral immune response against simian immunodeficiency virus (SIVmac251), 11 macaques were immunized with live-attenuated SIVmac239Deltanef either intravenously or via the tonsils and exposed to SIVmac251 after either 6 or 15 months along with unvaccinated controls. RESULTS: Independent of the route of vaccine application, viremia was significantly reduced in vaccinees compared with controls 2 weeks post-challenge. Concomitantly, viremia correlated inversely with SIV-specific IgG, complement-mediated lysis and neutralizing antibodies and these parameters seemed to contribute to reduced viremia. During chronic infection, six monkeys controlled viremia in the circulation (two or fewer infectious units per 10(6) PBMCs) and showed no signs of trapping in lymphatic tissues (Appendix S1). CONCLUSIONS: As no significant differences were observed throughout the study, with respect to the humoral immune response and viremia control, between the two vaccinated cohorts, mucosal immunization strategies are recommended due to more simplified application. PMID- 20015160 TI - Acute coronary syndromes in treatment-naive black South africans with human immunodeficiency virus infection. AB - BACKGROUND: HIV patients on protease inhibitors have greater risk of acute coronary syndromes (ACS) but little is known about treatment-naive patients. METHODS AND RESULTS: Authors conducted a prospective single-center study from Soweto, South Africa, comparing the clinical and angiographic features of treatment-naive HIV positive and negative patients with ACS. Between March 2004 and February 2008, 30 consecutive treatment-naive HIV patients with ACS were compared to the next HIV-negative patient as a 1:1 control. HIV patients were younger (43 +/- 7 vs. 54 +/- 13, P = 0.004) and, besides smoking (73% vs. 33%, P = 0.002), had fewer risk factors than the control group with less hypertension (23% vs. 77%, P = 0.0001), diabetes (3% vs. 23%, P = 0.05), LDL hyperlipidemia (2.2 +/- 0.9 vs. 3.0 +/- 1.2, P = 0.006), and other coronary risk factors (7% vs. 53%, P = 0.0001). HDL was lower in the HIV group (0.8 +/- 0.3 vs. 1.1 +/- 0.4, P = 0.001). Atherosclerotic burden was lower in the HIV group with more normal infarct-related arteries (47% vs. 13%, P = 0.005) but a higher degree of large thrombus burden (43% vs. 17%, P = 0.02). Stents were used to a similar degree in HIV and control patients (30% vs. 37%, P = 0.78) with more target lesion revascularization in the HIV group (56% vs. 0%, P = 0.008). CONCLUSION: Treatment naive HIV patients with ACS are younger and have fewer traditional risk factors than HIV-negative patients. HIV patients have less atherosclerotic but higher thrombotic burden which may imply a prothrombotic state in the pathogenesis of ACS in these patients. PMID- 20015161 TI - Living 'a life like ours': support workers' accounts of substitute decision making in residential care homes for adults with intellectual disabilities. AB - In England and Wales, the Mental Capacity Act 2005 (MCA) provides a new legal framework to regulate substitute decision-making relating to the welfare of adults who lack the capacity to make one or more autonomous decisions about their care and support. Any substitute decision made on behalf of an adult lacking capacity must be in his/her 'best interests'. However, the value of adopting established principles and procedures for substitute decision-making in practice is uncertain, and little is known about the legal or ethical dynamics of social care support, including the day-to-day residential support provided to adults with intellectual disabilities (ID). Methods This paper reports a qualitative, grounded theory analysis of 21 interviews with support workers working in residential care homes for adults with ID, and observations of care practices. Results In contrast to the narrow legal responsibilities placed upon them, it is argued that support workers interpret substitute decision-making within a broad moral account of their care role, orientating their support towards helping residents to live 'a life like ours'. In so doing, support workers describe how they draw on their own values and life experiences to shape the substitute decisions that they make on behalf of residents. Conclusions Support workers' accounts reveal clear discrepancies between the legal regulation of substitute decision-making and the ways that these support workers make sense of their work. Such discrepancies have implications both for the implementation of the MCA, and for the role of support workers' values in the conceptualisation and delivery of 'good' care. PMID- 20015162 TI - A study of PCR inhibition mechanisms using real time PCR. AB - In this project, real time polymerase chain reaction (PCR) was utilized to study the mechanism of PCR inhibition through examination of the effect of amplicon length, melting temperature, and sequence. Specifically designed primers with three different amplicon lengths and three different melting temperatures were used to target a single homozygous allele in the HUMTH01 locus. The effect on amplification efficiency for each primer pair was determined by adding different concentrations of various PCR inhibitors to the reaction mixture. The results show that a variety of inhibition mechanisms can occur during the PCR process depending on the type of co-extracted inhibitor. These include Taq inhibition, DNA template binding, and effects on reaction efficiency. In addition, some inhibitors appear to affect the reaction in more than one manner. Overall we find that amplicon size and melting temperature are important in some inhibition mechanisms and not in others and the key issue in understanding PCR inhibition is determining the identity of the interfering substance. PMID- 20015163 TI - Analysis of pigmented inkjet printer inks and printed documents by laser desorption/mass spectrometry. AB - Anyone with a computer, scanner, and color printer has the capability for creating documents such as identification cards, passports, and counterfeit currency. Laser desorption mass spectrometry (LDMS) has been demonstrated as a powerful tool for colorant analysis. Inkjet printers are now moving largely toward the use of pigments as colorants; their insolubility makes analysis by simpler methods such as thin-layer chromatography no longer an option. Recent developments in pigmented inkjet printer inks, such as gloss optimizers that coat pigment particles, may prohibit colorant analysis by LDMS. We demonstrate here that pigments used in inks from two Epson printers can be detected and analyzed by LDMS. Also, LDMS spectra of various colors created using a 4-cartridge (cyan/magenta/yellow/black, CMYK) inkset are evaluated, to begin to develop an approach for unraveling LDMS data from real samples, to determine the number of inks used by a printer, and the chemical composition of the colorants. PMID- 20015164 TI - Stature estimation from foot length using universal regression formula in a North Indian population. AB - Stature is a significant parameter in establishing identity of an unknown. Conventionally, researchers derive regression formula separately for males and females. Sex, however, may not always be determined accurately, particularly in dismembered remains and thus the need for a universal regression formula for stature estimation irrespective of sex of an individual. The study was carried out in an endogamous group of North India to compare the accuracy of sex-specific regression models for stature estimation from foot length with the models derived when the sex was presumed as unknown. The study reveals that regression equation derived for the latter can estimate stature with reasonable accuracy. Thus, stature can be estimated accurately from foot length by regression analysis even when sex remains unknown. PMID- 20015165 TI - Forensic analysis of explosives using isotope ratio mass spectrometry (IRMS)- part 2: forensic inter-laboratory trial: bulk carbon and nitrogen stable isotopes in a range of chemical compounds (Australia and New Zealand). AB - Comparability of data over time and between laboratories is a key issue for consideration in the development of global databases, and more broadly for quality assurance in general. One mechanism that can be utilized for evaluating traceability is an inter-laboratory trial. This paper addresses an inter laboratory trial conducted across a number of Australian and New Zealand isotope ratio mass spectrometry (IRMS) laboratories. The main objective of this trial was to determine whether IRMS laboratories in these countries would record comparable values for the distributed samples. Four carbon containing and four nitrogen containing compounds were distributed to seven laboratories in Australia and one in New Zealand. The laboratories were requested to analyze the samples using their standard procedures. The data from each laboratory was evaluated collectively using International Standard ISO 13528 (Statistical methods for use in proficiency testing by inter-laboratory comparisons). "Warning signals" were raised against one participant in this trial. "Action signals" requiring corrective action were raised against four participants. These participants reviewed the data and possible sources for the discrepancies. This inter laboratory trial was successful in providing an initial snapshot of the potential for traceability between the participating laboratories. The statistical methods described in this article could be used as a model for others needing to evaluate stable isotope results derived from multiple laboratories, e.g., inter-laboratory trials/proficiency testing. Ongoing trials will be conducted to improve traceability across the Australian and New Zealand IRMS community. PMID- 20015166 TI - Forensic analysis of explosives using isotope ratio mass spectrometry (IRMS)- part 1: instrument validation of the DELTAplusXP IRMS for bulk nitrogen isotope ratio measurements. AB - A significant amount of research has been conducted into the use of stable isotopes to assist in determining the origin of various materials. The research conducted in the forensic field shows the potential of isotope ratio mass spectrometry (IRMS) to provide a level of discrimination not achievable utilizing traditional forensic techniques. Despite the research there have been few, if any, publications addressing the validation and measurement uncertainty of the technique for forensic applications. This study, the first in a planned series, presents validation data for the measurement of bulk nitrogen isotope ratios in ammonium nitrate (AN) using the DELTA(plus)XP (Thermo Finnigan) IRMS instrument equipped with a ConFlo III interface and FlashEA 1112 elemental analyzer (EA). Appropriate laboratory standards, analytical methods and correction calculations were developed and evaluated. A validation protocol was developed in line with the guidelines provided by the National Association of Testing Authorities, Australia (NATA). Performance characteristics including: accuracy, precision/repeatability, reproducibility/ruggedness, robustness, linear range, and measurement uncertainty were evaluated for the measurement of nitrogen isotope ratios in AN. AN (99.5%) and ammonium thiocyanate (99.99+%) were determined to be the most suitable laboratory standards and were calibrated against international standards (certified reference materials). All performance characteristics were within an acceptable range when potential uncertainties, including the manufacturer's uncertainty of the technique and standards, were taken into account. The experiments described in this article could be used as a model for validation of other instruments for similar purposes. Later studies in this series will address the more general issue of demonstrating that the IRMS technique is scientifically sound and fit-for-purpose in the forensic explosives analysis field. PMID- 20015167 TI - Intellectual quotient of juveniles evaluated in a forensic psychiatry clinic after committing a violent crime. AB - The purpose of this preliminary study is to evaluate if there is a difference between the intelligence quotient (IQ) of 27 adolescent defendants referred to the Bellevue Hospital Center Forensic Psychiatry Clinic after committing violent crimes, and those adolescents in the same age group in the general population of the United States, as defined by the norms of the psychometric testing instrument Wechsler Intelligence Scale for Children, 4th edition (WISC-IV). The IQ scores and sub-scores were compared to IQ scores of the general population (mean = 100, SD = 15) using a Z-test. The mean for the Full Scale IQ was 82.93. The means for the subtests which include Processing Speed Index, Perceptual Reasoning Index, Verbal Comprehension Index, and Working Memory Index, were: 78.48, 87.78, 86.70 (p < 0.05), and 90.78 (p = 0.09) respectively. There is a statistically significant difference in the IQ scores of the violent juveniles studied when compared to the general population. PMID- 20015168 TI - Prevalence of intellectual disabilities in Norway: Domestic variance. AB - Based on national registers, the prevalence of intellectual disability (ID) in Norway is estimated to be 0.44 per 100 inhabitants. This study aimed to examine geographic and urban-rural differences in the prevalence of ID in Norway. Methods A survey based on the national register. Financial transfers intended to provide equal services to people with ID are based on these reports. Results A higher prevalence was found in the North region of Norway. A negative correlation between the population density and the prevalence of ID was also found. Conclusion There was considerable geographic and urban-rural differences in the prevalence of ID, which may be attributable to not only the large diversity of services, but also some other factors. The results were discussed with respect to the deinstitutionalisation progress, resource-intensive services and costs. Differences also reflect some problems in diagnosing ID in people having mild ID. PMID- 20015169 TI - Evaluation of JPEG and JPEG2000 compression algorithms for dermatological images. AB - BACKGROUND: Some image compression methods are used to reduce the disc space needed for the image to store and transmit the image efficiently. JPEG is the most frequently used algorithm of compression in medical systems. JPEG compression can be performed at various qualities. There are many other compression algorithms; among these, JPEG2000 is an appropriate candidate to be used in future. OBJECTIVE: To investigate perceived image quality of JPEG and JPEG2000 in 1 : 20, 1 : 30, 1 : 40 and 1 : 50 compression rates. METHODS: In total, photographs of 90 patients were taken in dermatology outpatient clinics. For each patient, a set which is composed of eight compressed images and one uncompressed image has been prepared. Images were shown to dermatologists on two separate 17-inch LCD monitors at the same time, with one as compressed image and the other as uncompressed image. Each dermatologist evaluated 720 image couples in total and defined whether there existed any difference between two images in terms of quality. If there was a difference, they reported the better one. Among four dermatologists, each evaluated 720 image couples in total. RESULTS: Quality rates for JPEG compressions 1 : 20, 1 : 30, 1 : 40 and 1 : 50 were 69%, 35%, 10% and 5% respectively. Quality rates for corresponding JPEG2000 compressions were 77%, 67%, 56% and 53% respectively. CONCLUSION: When JPEG and JPEG2000 algorithms were compared, it was observed that JPEG2000 algorithm was more successful than JPEG for all compression rates. However, loss of image quality is recognizable in some of images in all compression rates. PMID- 20015170 TI - Psoriasis is independently associated with psychiatric morbidity and adverse cardiovascular risk factors, but not with cardiovascular events in a population based sample. AB - BACKGROUND: Psoriasis may significantly reduce quality of life. Previous studies reported an association of psoriasis and cardiovascular risk factors and cardiovascular events. The extent to which psoriasis is associated with psychiatric morbidity and the role of psychiatric comorbidity as a potential confounder of the association between psoriasis and cardiovascular morbidity require further investigation. OBJECTIVES: To study the association between psoriasis, psychiatric morbidity and cardiovascular morbidity. METHODS: Case control study utilizing an interdisciplinary administrative outpatient database from Germany. Patients with confirmed diagnosis of prevalent psoriasis within the study period (2003-2004) (n = 3147, mean age 57 years) were individually matched for age and gender with 3147 controls without psoriasis. The relationship of psoriasis with psychiatric morbidities (depression, stress-related disorders, behaviour disorders and schizophrenic disorders), cardiovascular risk factors (diabetes, hypertension, obesity and dyslipidaemia) and cardiovascular events [myocardial infarction (MI), stroke] was investigated using logistic and linear regression models. RESULTS: Crude analyses suggested an association of psoriasis with depression, stress-related disorders, behaviour disorders and cardiovascular risk factors, but not with MI [odds ratio (OR) 1.14; 95% confidence interval (95% CI) 0.81-1.62] or stroke (OR 0.97; 95% CI 0.61-1.54). Multivariate models controlling for age, gender and consulting behaviour indicated that psoriasis is independently associated with depression (OR 1.49; 95% CI 1.20-1.86), stress related disorders (OR 1.41; 95% CI 1.22-1.62), behaviour disorders (OR 1.58; 95% CI 1.05-2.39), diabetes (OR 1.21 95% CI 1.04-1.40), hypertension (OR 1.34; 95% CI 1.18-1.51), dyslipidaemia (OR 1.29; 95% CI 1.07-1.55), and obesity (OR 1.63; 95% CI 1.39-1.90). For each psychiatric condition, the likelihood of being affected significantly increased with each physician visit due to psoriasis, suggesting that the risk of psychiatric comorbidity increases with the severity of psoriasis. CONCLUSION: Psoriasis appears to be independently associated with major psychiatric disorders and with cardiovascular risk factors, but not with cardiovascular events. PMID- 20015171 TI - Authors' willingness for second-opinion teledermatology in difficult-to-manage cases: 'an online survey'. AB - BACKGROUND: Expert non-availability for an instant difficult-to-manage case may limit the online discussion group to provide second-opinion teledermatology for registered dermatologists. Author-based second-opinion teledermatology (AST) is proposed and assessment of authors' response to answer queries and their willingness to participate in teledermatology consultation is analysed. MATERIALS AND METHODS: Authors of 292 articles from seven non-open access dermatology and three general medical journals addressing difficult-to-manage cases from June 2007 to December 2008 participated in this online dermatology authors' survey. Authors' response to reprint requests, answer literature queries and willingness to offer second-opinion teledermatology were the main outcome measures. Perceived views were also documented. RESULTS: The scores for reprints requests vs. received were (chi(2) = 13.57, n = 292), P = 0.000. The scores for articles read, analysed and literature queries sent vs. authors answered were (chi(2) = 11.09, n = 190), P = 0.001. The scores for authors responded to reprints vs. willing to offer second-opinion teledermatology consultation were (chi(2) = 5.65, n = 190), P = 0.017 and authors responded to reprint requests, answered queries and willing to offer second-opinion (chi(2) = 5.65, n = 190), P = 0.017. CONCLUSION: A dermatologist obtains literature, answered queries and constructive suggestions with additional informative material and pools second-opinions from the authors using their e-mail to deliver quality health care. AST is an alternative to online discussion group for second-opinion teledermatology practice in difficult to-manage cases. Uniform international laws are to be framed to facilitate telemedicine care for a difficult-to-manage case. PMID- 20015172 TI - Sexually transmitted infections and related sociodemographic factors in Lisbon's major Venereology Clinic: a descriptive study of the first 4 months of 2007. AB - OBJECTIVES: To assess the prevalence of sexually transmitted infections (STI) and relationships between them and sociodemographic/behavioural data in the major Venereology Clinic in Lisbon. METHODS: Every patient attending this STI clinic for the first time in the first 17 weeks of 2007 was enrolled in this study. Early syphilis, Chlamydia trachomatis infection, gonorrhoea, human immunodeficiency virus (HIV) first diagnosed and genital warts were considered for the study of statistical associations with sociodemographic/behavioural variables or other STI. Data were analysed with an exact significance level of 5%. RESULTS: A total of 743 patients were included. In women (n = 296), the only significant associations found were for C. trachomatis and being non-Caucasian (OR = 2.13, CI 1.17-3.9) or being younger than 25 years (OR = 1.9, CI 1.31-2.79). Men who have sex with men (176 of the 447 male patients) contributed to 39% of the STI, although more than half of the early syphilis, gonorrhoea and HIV cases were diagnosed in this group. CONCLUSIONS: Despite its limitations, our study allows some insight into the relationships between sociodemographic factors and STI in a selected population attending Lisbon's major Venereology Clinic. The results were similar to those of other European studies and to recent trends in STI, but differ in the association between C. trachomatis infection and non Caucasian women, which is similar to cities with significant African minorities. Attention should be given to the associations found between men who have sex with men and STI such as gonorrhoea, syphilis and HIV, as these associations can lead to serious difficulties in the control of HIV infection in Portugal. PMID- 20015173 TI - Dermoscopic features of Bowen's disease in Asians. AB - BACKGROUND: Previously, dermoscopic features of Bowen's disease (BD) were extensively investigated in two studies, but there were some discrepancies. The dispute necessitated a further study concerning the dermoscopic features of BD. OBJECTIVE: To describe the dermoscopic features of BD in Asians and to assess dermoscopy as a post-treatment monitoring tool of BD. MATERIALS AND METHODS: Dermoscopic examinations of histopathologically diagnosed 26 BD lesions were performed to evaluate for the presence of various dermoscopic features. In addition, the correlating changes of dermoscopic features and histopathological results before and after treatments were assessed in five patients with BD. RESULTS: Dermoscopically, 10 lesions were pigmented and 16 lesions were non pigmented. The most frequent dermoscopic findings of BD were vascular structures (96%) and a scaly surface (96%). Among vascular structure, glomerular vessels were most frequently observed (77%). The other vascular structures in our study were linear irregular vessels, dotted vessels, polymorphous/atypical vessels and arborizing vessels. Among five patients who had been treated with either photodynamic therapy or 5% imiquimod cream, four patients revealed disappearance of dermoscopic vascular structures, but one patient showed remaining vascular structures after treatment. Skin biopsy from treated lesions disclosed clearance of BD in four patients who had no vascular structures but remaining BD in the patient whose dermoscopic finding displayed no disappearance of vascular structures. CONCLUSIONS: Vascular structures, especially glomerular vessels plus a scaly surface, were common dermoscopic findings of BD in Asians. In addition, existence of dermoscopic vascular structures after treatment appears to be associated with residual disease. PMID- 20015174 TI - Genital verrucous carcinoma is associated with lichen sclerosus: a retrospective study and review of the literature. AB - BACKGROUND: The association of lichen sclerosus (LS) with genital squamous cell carcinoma is well recognized. However, the relationship between LS and verrucous carcinoma remains unclear. OBJECTIVE: To evaluate the associations of genital and perianal verrucous carcinomas with LS. METHODS: We conducted a retrospective study on patients with a genital or perianal verrucous carcinoma and reviewed their histopathology specimens and clinical notes. We also conducted a literature review. RESULTS: We identified a total of 13 patients (including 6 women and 7 men) with a genital or perianal verrucous carcinoma. All 5 women with vulval verrucous carcinoma had coexisting LS (5/5), and 1 man with penile verrucous carcinoma had coexisting LS (1/3). In contrast, no coexisting LS was found in all 5 cases of perianal verrucous carcinoma (0/5). Half of the cases of verrucous carcinoma with coexisting LS had recurrences (3/6), while no recurrences were found in those without coexisting LS (0/7). CONCLUSIONS: Our study and review of the literature demonstrate that vulval verrucous carcinoma is strongly associated with LS. In contrast, perianal verrucous carcinoma is not associated with LS. When genital verrucous carcinoma is diagnosed, it is important to consider LS as a potential concomitant diagnosis and offer appropriate treatments and close follow-up to detect recurrence of verrucous carcinoma. PMID- 20015176 TI - Cutaneous sarcoidosis: updates in the pathogenesis. AB - Sarcoidosis is a multiorgan granulomatous disease in which the skin is one of the frequently involved target organs. Cutaneous involvement occurs in a third of patients with sarcoidosis and has protean manifestations. More than a century has passed since the initial description of sarcoidosis, but its cause continues to be an enigma. Recent studies have introduced several new insights into the pathogenesis of this disease. The aim of this literature review was to provide a comprehensive overview on the current updates in the pathogenesis of sarcoidosis. This review has revealed that several genetic polymorphisms are associated with an increased risk of developing sarcoidosis, suggesting that genetic susceptibility to sarcoidosis is probably polygenic. Environmental factors may also modify the susceptibility to sarcoidosis. Evidence favouring an infectious aetiology has been accumulating, but the results of studies are conflicting. The current concept is that the pathogenesis of sarcoidosis involves a T-helper-1 mediated immune response to environmental antigens in a genetically susceptible host. The studies carried out on sarcoidosis have largely focused on the pulmonary aspects and have been mainly conducted by respiratory physicians. In contrast, research conducted on the cutaneous aspects of sarcoidosis is comparatively limited. Although tremendous advances have been made, there is a significant gap between the vast knowledge accumulated on sarcoidosis in recent years and the understanding of this disease. PMID- 20015177 TI - Microbiological analysis of epidermal growth factor receptor inhibitor therapy associated paronychia. AB - BACKGROUND: Paronychia is a well-known, but difficult to treat cutaneous toxicity associated with epidermal growth factor receptor (EGFR) inhibitor therapy. Although bacterial and fungal infections as well as mechanical trauma may play a role as co-pathogens, there is no good basis for an empirical antimicrobial chemotherapy in these patients. MATERIALS AND METHODS: We retrospectively analysed the microbiological results and resistance analysis of 42 cases of EGFR inhibitor-associated paronychia induced by cetuximab. RESULTS: We identified 20 different species, among these 72% Gram-positive bacteria, 23% Gram-negative bacteria and 5%Candida species. About half of the microbes identified may be considered as residential bacterial flora of the skin, but isolation of microbes from paronychia may indicate a pathogenic relevance for this type of reaction. Eight of our patients were treated with oral antibiotics, whereas two patients received oral antimycotic therapy. All other cases of paronychia were controlled using topical antiseptic, antibiotic and antimycotic agents. CONCLUSION: Empirical oral antibiotic treatment may be performed with oral cephalosporines, ciprofloxacin, levofloxacin or moxifloxacin, as these antimicrobials have high in vitro activity against the majority of the isolated microorganisms and reach high concentrations in the relevant tissue. PMID- 20015178 TI - Caveolae in Fabry telangiectagia. PMID- 20015179 TI - A novel deletion mutation in P2RY5/LPA(6) gene cause autosomal recessive woolly hair with hypotrichosis. PMID- 20015180 TI - Who uses sunbeds? A systematic literature review of risk groups in developed countries. AB - Skin cancer is caused by ultraviolet radiation (UVR). Indoor tanning is a totally avoidable risk behaviour. This review addresses the specific characteristics of sunbed users and the differences in motivation and risk perception compared with non-users. This review is based solely on empirical original articles. Based on literature searches with widely used reference databases ('PubMed', 'OVID', 'Social Citation Index', 'ERIC--Educational Resources Information Center', 'Web of Science' and the 'International Bibliography of the Social Sciences'), we included studies from developed nations with a publication date between 1 January 2000 and 12 August 2008. All studies were selected, classified and coded simultaneously by both authors on a blinded basis. All searches were performed on 13 and 14 August 2008. In accordance with the QUOROM and the MOOSE Statements, we identified 16 original studies. The typical sunbed user is female, between 17 and 30 years old, and tends to live a comparatively unhealthy lifestyle: Users smoke cigarettes and drink alcohol more frequently and eat less healthy food than non users. Users are characterized by a lack of knowledge about health risks of UVR, and prompted by the frequent use of sunbeds by friends or family members and the experience of positive emotions and relaxation by indoor tanning. This review is the first systematic review on risk groups among sunbed users that has been published in a scientific journal. There is still a lack of information among users, particularly among young people regarding the safety of solariums. PMID- 20015181 TI - Ineffectiveness of sun awareness posters in dermatology clinics. AB - BACKGROUND: Although sun awareness posters have been used in doctors' offices and clinics for decades to promote sun protective behaviour, there is no evidence of their usefulness. OBJECTIVES: To investigate whether sun awareness posters lead to inquiry of skin cancer and sun protection measures. METHOD: Patients considered at risk for skin cancer seen at a dermatology clinic were randomly asked to complete a questionnaire designed to assess the effectiveness of three different sun awareness posters placed in patient rooms. The posters were selected on the basis of their catchy slogan and eye-appealing images, and included those featuring parental interest, sex appeal and informative advice. RESULTS: Only half of the patients noticed the posters (50.6%). The poster with sex appeal garnered the most attention (67.8%), followed by the informative poster (49.2%) and the parental interest poster (35.8%) (P < 0.001). Although patients who noticed the sun awareness poster inquired about cutaneous cancers and sun protection practices twice as often as those who did not notice the poster, only one-tenth of such inquiries were attributed to the poster ( approximately 5% of the target population). As reported in the questionnaire, the posters themselves were less effective than the advice of physicians in influencing patient attitudes towards sun protection measures. CONCLUSION: Organizations that produce and disseminate posters should consider beyond focus groups when they design their posters and should consider field testing their products to ensure that they are reaching the targeted audience and are having the expected beneficial effect, otherwise their posters are simply decorative. PMID- 20015182 TI - Hydrogenosomes of laboratory-induced metronidazole-resistant Trichomonas vaginalis lines are downsized while those from clinically metronidazole-resistant isolates are not. AB - Trichomonas vaginalis is the most common sexually transmitted protozoan in the world and its resistance to metronidazole is increasing. The purpose of this study was to demonstrate that clinical metronidazole resistance in T. vaginalis does not occur via the same mechanism as laboratory-induced metronidazole resistance--that is, via hydrogenosome down sizing. Ultrathin sections of this parasite were examined using transmission electron microscopy and the size and area of the cell and hydrogenosomes were compared between drug-resistant laboratory lines and clinically resistant isolates. Clinical metronidazole resistant T. vaginalis had similar-sized hydrogenosomes as a metronidazole sensitive isolate. Inducing metronidazole resistance in both of these isolates caused down sizing of hydrogenosomes. Inducing toyocamycin resistance did not cause any ultrastructural changes to the cell or to the hydrogenosome. No correlation between hydrogenosome number and the drug-resistant status of T. vaginalis isolates and lines was observed. This report demonstrates that clinical metronidazole resistance is not associated with down-sized hydrogenosomes, thus indicating that an alternative resistance mechanism is used by T. vaginalis. PMID- 20015183 TI - Characterization of acanthamoeba isolates from dust of a public hospital in Curitiba, Parana, Brazil. AB - Occurrence of Acanthamoeba in the hospital environment may represent a health risk for patients, since these organisms can cause severe opportunistic illness, such as keratitis, and also can harbor pathogenic agents. We analyzed the dust from some environments of a public hospital in Curitiba, Parana State, Brazil. Two distinct populations of Acanthamoeba were isolated in five locations and morphologically classified as group I and group II according to Pussard and Pons. Isolates were identified as Acanthamoeba by PCR using primers to amplify a region of 18S rDNA, which showed variation in the product length among the isolates. A cloned culture of group II showed greater growth at 37 degrees C and in media with 0.1, 0.5, and 1.0 M mannitol, which are the physiological characteristics of pathogenic Acanthamoeba. Monitoring the presence of Acanthamoeba in hospital units, as well as evaluating the pathogenicity of the isolates, can be an approach to alert the health professionals to improve the disinfection procedures and minimize the risks of treating this problematic disease caused by this protozoan. PMID- 20015184 TI - Variability of wax ester fermentation in natural and bleached Euglena gracilis Strains in response to oxygen and the elongase inhibitor flufenacet. AB - Euglena gracilis is able to synthesize adenosine triphosphate under anaerobic conditions through a malonyl-independent fatty acid synthesis leading to wax ester fermentation. Mitochondrial fatty acid synthesis uses acetyl-CoA and propionyl-CoA as C2- and C3-donors for de novo synthesis of even- and odd numbered fatty acids, respectively. Euglena's wax ester fermentation has only been described in the E. gracilis strain 1224-5/25 Z. Here we investigate eight E. gracilis strains isolated in 1932-1958 from different localities in Europe and two bleached substrains of E. gracilis 1224-5/25, obtained by treatment with streptomycin and ofloxacin, and examine their anaerobic growth, wax ester fermentation, and wax ester composition. Under ambient oxygen levels, all strains accumulated wax esters in concentrations between 0.3% and 3.5% of the dry weight, but the strains revealed marked differences in wax ester accumulation with respect to anaerobic growth. Most fermenting strains tested showed increased wax ester synthesis under anaerobic conditions as well as the increased synthesis of odd-numbered fatty acids and alcohols suggesting an activation of the mitochondrial fatty acid biosynthesis pathway. Addition of the elongase inhibitor flufenacet to the growth medium specifically reduced the accumulation of odd numbered fatty acids and alcohols and tended to increase the overall yield of anaerobic wax esters. PMID- 20015185 TI - Conserved meiotic genes point to sex in the choanoflagellates. AB - The choanoflagellates are a widespread group of heterotrophic aquatic nanoflagellates, which have recently been confirmed as the sister-group to Metazoa. Asexual reproduction is the only mode of cell division that has been observed within the group; at present the range of reproductive modes, as well as the ploidy level, within choanoflagellates are unknown. The recent discovery of long terminal repeat retrotransposons within the genome of Monosiga brevicollis suggests that this species also has sexual stages in its life cycle because asexual organisms cannot tolerate retrotransposons due to the rapid accumulation of deleterious mutations caused by their transposition. We screened the M. brevicollis genome for known eukaryotic meiotic genes, using a recently established "meiosis detection toolkit" of 19 genes. Eighteen of these genes were identified, none of which appears to be a pseudogene. Four of the genes were also identified in expressed sequence tag data from the distantly related Monosiga ovata. The presence of these meiosis-specific genes provides evidence for meiosis, and by implication sex, within this important group of protists. PMID- 20015186 TI - Morphology, ultrastructure, and small subunit rDNA phylogeny of the marine heterotrophic flagellate Goniomonas aff. amphinema. AB - Marine goniomonads have a worldwide distribution but ultrastructural information has not been available so far. An isolate of the heterotrophic marine nanoflagellate Goniomonas (G. aff. amphinema) from North Wales (UK) has been studied, providing information on its morphology and cellular structure using video, electron, laser scanning confocal microscopy (LSCM), and atomic force microscopy. Here, we describe a new feature, a granular area, potentially involved in particle capture and feeding. The binding of the lectin wheat germ agglutinin to the granular area of cells with discharged ejectisomes indicates the adhesive nature of this novel feature. The presence of a microtubular intracellular cytopharynx, apparently also used for feeding, has been revealed by LSCM. The small subunit rRNA gene of the isolate has been sequenced (1,788 bp). Phylogenetic results corroborate significant genetic divergence within the marine members of Goniomonas. This work highlights the need for integrated morphological, ultrastructural, and molecular investigation when describing and studying heterotrophic nanoflagellates. PMID- 20015187 TI - Psoriatic arthritis: a review. AB - Psoriatic arthritis (PsA) is an immunologically triggered, chronic inflammatory arthropathy, which can have a lasting influence on the quality of life of affected individuals. An early diagnosis is essential in order to institute adequate therapy. Both dermatologists and rheumatologists should be involved in the diagnosis and management of the disorder. Mild forms can be managed with NSAID and systemic corticosteroids. In acute forms with a confirmed diagnosis and oligo- to polyarticular involvement, disease-modifying therapy with DMARD (Disease-Modifying Antirheumatic Drugs) is indicated. New studies of PsA show that tumor necrosis factor (TNF) plays a central role in mediating inflammation. For this reason, TNF-alpha antagonists have become more and more important as a second-line therapy for PsA. PMID- 20015188 TI - Borrelia in granuloma annulare, morphea and lichen sclerosus: a PCR-based study and review of the literature. AB - BACKGROUND: Morphea, granuloma annulare (GA) and lichen sclerosus et atrophicans (LSA) have also been suggested to be linked to Borrelia infection. Previous studies based on serologic data or detection of Borrelia by immunohistochemistry and polymerase chain reaction (PCR) reported contradictory results. Thus, we examined skin biopsies of morphea, GA and LSA by PCR to assess the prevalence of Borrelia DNA in an endemic area and to compare our results with data in the literature. METHODS: Amplification of DNA sequences of Borrelia burgdorferi sensu lato by nested PCR from formalin-fixed and paraffin-embedded skin biopsies of morphea, GA and LSA, followed by automated sequencing of amplification products. PCR-based studies on Borrelia species in these disorders published until July 2009 were retrieved by a literature search. RESULTS: Borrelia DNA was detected in 3 of 112 skin biopsies (2.7%) including one of 49 morphea biopsies (2.0%), one of 48 GA biopsies (2.1%) and one of 15 LSA biopsies (6.6%). Amplification products belonged to B. burgdorferi sensu stricto in two cases available for sequence analysis. CONCLUSIONS: The results of our and most of other PCR-based studies do not argue for a significant association of B. burgdorferi sensu lato with morphea, GA, LSA. PMID- 20015189 TI - Formalin deposition as artifact in biopsies from patients affected by a new variant of endemic pemphigus foliaceus in El Bagre, Colombia, South America. AB - BACKGROUND: Most autoimmune diseases occur sporadically; however, endemic pemphigus foliaceus (EPF) is present in specific locales restricted to some geographic rural regions mostly in South America, Central America and in Tunisia (Africa). Its geographic restriction makes it an invaluable natural model for studying how the environment, genetic background and host response contribute to the development of autoimmunity. We described a new variant of EPF in El Bagre, Colombia, (El Bagre-EPF). When we examined the skin biopsies from 10 patients and controls from the endemic area, we detected in a systematic manner several types of pigmentation, sometimes intracellular, and sometimes in the extracellular matrix in most biopsies. AIM: We aim to determine the nature of this pigment in these skin biopsies. METHODS: We studied 10 patients and 10 controls matched by sex, age and work activity living in the endemic area by routine hematoxylin and eosin (H&E). RESULTS: We were unable to find any bacteriological or parasitic organism. Specifically, we searched for several tropical disease agents as possible causative agents of this pigment. Iron stains and melanin pigment bleaching techniques failed to determine the etiology of this pigment. We then tried the removal of formalin pigment using picric acid. The pigment was removed after very strong treatment with different acids including picric acid. CONCLUSIONS: Formalin pigment shares many properties with hemozoin. In this case, the authors recommend the use of neutral buffered formalin to prevent the formation of formalin pigment especially after long periods of fixation when taking biopsies under extreme temperature and environmental humidity. PMID- 20015190 TI - Melanocytic nevi in pregnancy: histologic features and Ki-67 proliferation index. AB - BACKGROUND: Changes in the clinical appearance of benign dermal nevi during pregnancy may be concerning for malignant transformation. Because the hormonal milieu of pregnancy has not proven to alter their banal behavior, histologic characterization is needed to prevent over-diagnosis and unnecessary treatment. METHODS: Dermal nevi excised from pregnant women (n = 16) were compared with nevi from location- and aged-matched control patients (n = 15). Histologic features and Ki-67 proliferation index were evaluated. RESULTS: Nevi in pregnancy were more likely to have dermal mitotic figures (62.5% vs. 13.3%, p = 0.028) and higher mitotic rates (1.44 vs. 0.20 mitoses/mm(2), p = 0.0027) than control nevi. A distinctive histologic entity, termed superficial micronodules of pregnancy (SMOPs), was observed more frequently in the nevi of pregnancy (81.3% vs. 26.7%, p = 0.040), and showed consistent immunoreactivity for HMB45. There was a trend toward higher Ki-67 proliferation index in the nevi of pregnancy (3.0% vs. 1.0%, p = 0.073). Prominent multinucleated melanocytes were seen only in controls. There was no significant difference in pigmentation or irritation changes between groups. CONCLUSIONS: Dermal nevi removed during pregnancy show characteristic histologic features including increased dermal mitoses, superficial micronodules of pregnancy (SMOPs), and trend toward increased Ki-67 proliferation index. PMID- 20015191 TI - Emotional and behavioural difficulties in young children with and without developmental delay: a bi-national perspective. AB - BACKGROUND: Relatively high rates of emotional and behavioural difficulties have been reported among children with intellectual or developmental disabilities. Little is known about nature of the relationship between cognitive development and psychopathology in very young children. METHODS: Secondary analysis of data from the first two waves of the UK's Millennium Cohort Study (n = 11,389) and the first two waves of the Longitudinal Study of Australian Children (n = 4,606). RESULTS: Two- to three-year- old children with developmental delay show significantly higher rates of emotional and behavioural difficulties than their peers. These differences appear to reflect a general association between cognitive performance and emotional/behavioural difficulties in the bottom half of the ability spectrum and threshold-based discontinuities within the bottom 3% of the distribution. In the UK, but to a much lesser extent in Australia, higher rates of emotional and behavioural difficulties among children with developmental delay may be partially attributed to greater risk of exposure to adverse socio economic circumstances. CONCLUSIONS: The results highlight the potential value of targeted preventative interventions for young children with developmental delay. PMID- 20015192 TI - Developmental phenotypes and causal pathways in attention deficit/hyperactivity disorder: potential targets for early intervention? AB - Early intervention approaches have rarely been implemented for the prevention of attention deficit/hyperactivity disorder (ADHD). In this paper we explore whether such an approach may represent an important new direction for therapeutic innovation. We propose that such an approach is most likely to be of value when grounded in and informed by developmental models of the dynamic, complex and heterogeneous nature of the condition. First, we set out a rationale for early intervention grounded in the science of ADHD viewed through developmental models. Second, we re-examine the concept of disorder-onset from the perspective of developmental trajectories and phenotypes. Third, we examine potential causal pathways to ADHD with regard to originating risk, pathophysiological mediators, environmental moderators and developmental continuities. Finally, we explore the potential value of strategies for identifying young children at risk for ADHD, and implementing interventions in ways that can target these underlying pathogenic processes. The utility of such an approach represents an important area for future research but still requires 'proof of concept'. Therefore prior to widespread clinical implementation, far greater knowledge is required of (i) developmental pathways into ADHD, (ii) the value of identifying neuropsychological mediators of these pathways, and (iii) the extent to which targeting mediating mechanisms will improve treatment outcomes for children with ADHD. PMID- 20015193 TI - Five- to six-year outcome and its prediction for children with ODD/CD treated with parent training. AB - BACKGROUND: While short-term effects of parent training (PT) have been extensively evaluated, long-term outcome and present predictors of a diagnosis for children with ODD/CD treated with parent training are very limited. METHOD: In the present study, diagnostic status as outcome and predictors of treatment response were examined in a 5-6-year follow-up. Out of 99 children who had been treated in a randomised controlled trial evaluating the effects of The Incredible Years parent training (PT) or combined parent training and child treatment (PT+CT) programme, 54.5% participated in the 5-6-year follow-up study. Their diagnostic status was determined with the Kiddie-SADS interview. RESULTS: While all children qualified for a diagnosis of ODD/CD before treatment, 5-6 years later, two-thirds no longer received such a diagnosis, the same proportion as found at the 1-year follow-up. The most powerful pre-treatment predictors of diagnostic status at the 5-6-year follow-up were living with mother only and being a girl. At post-treatment the most powerful predictor was found to be high levels of child externalising problems. CONCLUSION: The findings of the study support the maintenance of positive long-term results for young children treated with parent training because of serious conduct problems, and identify characteristics of children and families in need of added support to parent training programmes. PMID- 20015194 TI - A novel pathway for receptor-mediated post-translational activation of inducible nitric oxide synthase. AB - Inducible nitric oxide synthase (iNOS) is a major source of nitric oxide during inflammation whose activity is thought to be controlled primarily at the expression level. The B1 kinin receptor (B1R) post-translationally activates iNOS beyond its basal activity via extracellular signal regulated kinase (ERK) mediated phosphorylation of Ser(745) . Here we identified the signalling pathway causing iNOS activation in cytokine-treated endothelial cells or HEK293 cells transfected with iNOS and B1R. To allow kinetic measurements of nitric oxide release, we used a sensitive porphyrinic microsensor (response time = 10 msec.; 1 nM detection limit). B1Rs signalled through Galphai coupling as ERK and iNOS activation were inhibited by pertussis toxin. Furthermore, transfection of constitutively active mutant Galphai Q204L but not Galphaq Q209L resulted in high basal iNOS-derived nitric oxide. G-betagamma subunits were also necessary as transfection with the beta-adrenergic receptor kinase C-terminus inhibited the response. B1R-dependent iNOS activation was also inhibited by Src family kinase inhibitor PP2 and trans-fection with dominant negative Src. Other ERK-MAP kinase members were involved as the response was inhibited by dominant negative H-Ras, Raf kinase inhibitor, ERK activation inhibitor and MEK inhibitor PD98059. In contrast, PI3 kinase inhibitor LY94002, calcium chelator 1,2-bis-(o-Aminophenoxy) ethane-N,N,N',N'-tetraacetic acid, tetraacetoxymethyl ester (BAPTA-AM), protein kinase C inhibitor calphostin C and protein kinase C activator PMA had no effect. Angiotensin converting enzyme inhibitor enalaprilat also directly activated B1Rs to generate high output nitric oxide via the same pathway. These studies reveal a new mechanism for generating receptor-regulated high output nitric oxide in inflamed endothelium that may play an important role in the development of vascular inflammation. PMID- 20015195 TI - The kinetochore protein Cenp-F is a potential novel target for zoledronic acid in breast cancer cells. AB - The anti-resorptive agent zoledronic acid inhibits key enzymes in the mevalonate pathway, disrupting post-translational modification and thereby correct protein localization and function. Inhibition of prenylation may also be responsible for the reported anti-tumour effects of zoledronic acid, but the specific molecular targets have not been identified. Cenp-F/mitosin, a kinetochore-associated protein involved in the correct separation of chromosomes during mitosis, has been shown to undergo post-translational prenylation and may therefore be a novel target contributing to the anti-tumour effects of zoledronic acid. We investigated whether zoledronic acid causes loss of Cenp-F from the kinetochore in breast cancer cells, to determine if the reported anti-tumour effects may be mediated by impairing correct chromosome separation. MDA-MB-436, MDA-MB-231 and MCF-7 breast cancer cells and MCF-10A non-malignant breast epithelial cells were treated with zoledronic acid in vitro, and the effect on Cenp-F localization was analysed by immunoflourescence microscopy. Zoledronic acid caused loss of Cenp-F from the kinetochore, accompanied by an increase in the number of cells in pro-, /prometa- and metaphase in all of the cancer cell lines. There was also a significant increase in the number of lagging chromosomes in mitotic cells. The effects of zoledronic acid could be reversed by inclusion of an intermediary of the mevalonate pathway, showing that the loss of Cenp-F from the kinetochore was caused by the inhibition of farnesylation. In contrast, no effect was seen on Cenp-F in non-malignant MCF-10A cells. This is the first report showing a specific effect of zoledronic acid on a protein involved in the regulation of chromosome segregation, identifying Cenp-F as a potential new molecular target for NBPs in tumour cells. PMID- 20015197 TI - Low-dose taxotere enhances the ability of sorafenib to induce apoptosis in gastric cancer models. AB - Despite the low efficacy of conventional antitumour drugs, chemotherapy remains an essential tool in controlling advanced gastric and oesophageal cancers. We aimed to provide a biological rationale based on the sorafenib-taxotere interaction for the clinical treatment of gastric cancer. In vitro experiments were performed on four human gastric cancer cell lines (GK2, AKG, KKP and NCI N87). Cytotoxicity was evaluated by sulforhodamine B (SRB) assay, cell cycle perturbations, apoptosis and mitotic catastrophe were assessed by flow cytometric and microscopic analyses, and protein expression was studied by Western blot. In the in vivo experiments, nude mice xenografted with the most resistant line were treated with sorafenib and docetaxel singly or in association. Sorafenib inhibited cell growth (IG(50) values ranged from 3.4 to 8.1 MUM) and caused down regulation of MAP-K/ERK phosphorylation and of mcl-1 and p-bad expression after a 48-hr exposure. Apoptosis induction was associated with caspase-3 and -9 activation and mitochondrial membrane depolarization. The drug combination enhanced apoptosis (up to 80%) and produced a synergistic interaction when low doses of the taxane preceded administration of the antityrosine kinase. This synergism was probably due to the induction of an anomalous multidiploid G0-G1 peak and to consequent mitotic catastrophe, which increased sensitivity to sorafenib. Consistent with in vitro results, the docetaxel-sorafenib sequence exhibited high therapeutic efficacy in NCI-N87 mouse xenografts producing tumour weight inhibition (> 65%), tumour growth delay (up to 25 days) and increased mouse survival (30%). Our findings suggest the potential clinical usefulness of treatment with sorafenib and docetaxel for advanced gastric cancer. PMID- 20015196 TI - Tumour hypoxia induces a metabolic shift causing acidosis: a common feature in cancer. AB - Maintenance of cellular pH homeostasis is fundamental to life. A number of key intracellular pH (pHi) regulating systems including the Na(+)/H(+) exchangers, the proton pump, the monocarboxylate transporters, the HCO(3)(-) transporters and exchangers and the membrane-associated and cytosolic carbonic anhydrases cooperate in maintaining a pHi that is permissive for cell survival. A common feature of tumours is acidosis caused by hypoxia (low oxygen tension). In addition to oncogene activation and transformation, hypoxia is responsible for inducing acidosis through a shift in cellular metabolism that generates a high acid load in the tumour microenvironment. However, hypoxia and oncogene activation also allow cells to adapt to the potentially toxic effects of an excess in acidosis. Hypoxia does so by inducing the activity of a transcription factor the hypoxia-inducible factor (HIF), and particularly HIF-1, that in turn enhances the expression of a number of pHi-regulating systems that cope with acidosis. In this review, we will focus on the characterization and function of some of the hypoxia-inducible pH-regulating systems and their induction by hypoxic stress. It is essential to understand the fundamentals of pH regulation to meet the challenge consisting in targeting tumour metabolism and acidosis as an anti-tumour approach. We will summarize strategies that take advantage of intracellular and extracellular pH regulation to target the primary tumour and metastatic growth, and to turn around resistance to chemotherapy and radiotherapy. PMID- 20015199 TI - Temporal relationship of autophagy and apoptosis in neurons challenged by low molecular weight beta-amyloid peptide. AB - Alzheimer's disease (AD) is an aging-related progressive neurodegenerative disorder. Previous studies suggested that various soluble Abeta species are neurotoxic and able to activate apoptosis and autophagy, the type I and type II programmed cell death, respectively. However, the sequential and functional relationships between these two cellular events remain elusive. Here we report that low molecular weight Abeta triggered cleavage of caspase 3 and poly (ADP ribose) polymerase to cause neuronal apoptosis in rat cortical neurons. On the other hand, Abeta activated autophagy by inducing autophagic vesicle formation and autophagy related gene 12 (ATG12), and up-regulated the lysoso-mal machinery for the degradation of autophagosomes. Moreover, we demonstrated that activation of autophagy by Abeta preceded that of apoptosis, with death associated protein kinase phosphorylation as the potential molecular link. More importantly, under Abeta toxicity, neurons exhibiting high level of autophagosome formation were absent of apoptotic features, and inhibition of autophagy by 3-methylade-nine advanced neuronal apoptosis, suggesting that autophagy can protect neurons from Abeta-induced apoptosis. PMID- 20015198 TI - Clinical outcomes in non-small-cell lung cancer patients with EGFR mutations: pooled analysis. AB - Non-small-cell lung cancer (NSCLC) with mutations in the epidermal growth factor receptor (EGFR) is a distinct subgroup of NSCLCs that is particularly responsive to EGFR tyrosine-kinase inhibitors (TKIs). A weighted pooled analysis of available studies was performed to evaluate clinical outcome in patients with EGFR-mutated NSCLC who were treated with chemotherapy or EGFR TKIs. Median progression-free survival (PFS) times were pooled from prospective or retrospective studies that evaluated chemotherapy or single-agent EGFR TKIs (erlotinib or gefitinib) in patients with NSCLC and EGFR mutations. Among the studies identified for inclusion in the analysis, 12 evaluated erlotinib (365 patients), 39 evaluated gefitinib (1069 patients) and 9 evaluated chemotherapy (375 patients). Across all studies, the most common EGFR mutations were deletions in exon 19 and the L858R substitution in exon 21. In the weighted pooled analysis, the overall median PFS was 13.2 months with erlotinib, 9.8 months with gefitinib and 5.9 months with chemotherapy. Using a two-sided permutation, erlotinib and gefitinib produced a longer median PFS versus chemotherapy, both individually (P= 0.000 and P= 0.002, respectively) and as a combined group (EGFR TKI versus chemotherapy, P= 0.000). EGFR TKIs appear to be the most effective treatment for patients with advanced EGFR-mutant NSCLC. Ongoing prospective trials comparing the efficacy of first-line chemotherapy and EGFR TKIs in EGFR mutant disease should provide further insight into the most appropriate way to treat this specific group of patients. PMID- 20015200 TI - TFPI-2 silencing increases tumour progression and promotes metalloproteinase 1 and 3 induction through tumour-stromal cell interactions. AB - Tissue factor pathway inhibitor-2 (TFPI-2) is a potent inhibitor of plasmin which activates matrix metalloproteinases (MMPs) involved in degradation of the extracellular matrix. Its secretion in the tumour microenvironment makes TFPI-2 a potential inhibitor of tumour invasion and metastasis. As demonstrated in aggressive cancers, TFPI-2 is frequently down-regulated in cancer cells, but the mechanisms involved in the inhibition of tumour progression remained unclear. We showed in this study that stable TFPI-2 down-regulation in the National Cancer Institute (NCI)-H460 non-small cell lung cancer cell line using specific micro interfering micro-interfering RNA promoted tumour progression in a nude mice orthotopic model that resulted in an increase in cell invasion. Moreover, TFPI-2 down-regulation enhanced cell adhesion to collagen IV and laminin via an increase in alpha(1) integrin on cell surface, and increased MMP expression (mainly MMP-1 and -3) contributing to cancer cell invasion through basement membrane components. This study also reveals for the first time that pulmonary fibroblasts incubated with conditioned media from TFPI-2 silencing cancer cells exhibited increased expression of MMPs, particularly MMP-1, -3 and -7, that are likely involved in lung cancer cell invasion through the surrounding stromal tissue, thus enhancing formation of metastases. PMID- 20015201 TI - Expression of NPP1 is regulated during atheromatous plaque calcification. AB - Mutations of the ENPP1 gene encoding ecto-nucleotide pyrophosphatase/phosphodiesterase 1 (NPP1) are associated with medial calcification in infancy. While the inhibitory role of matrix proteins such as osteopontin (OPN) with respect to atherosclerotic plaque calcification has been established, the role of NPP1 in plaque calcification is not known. We assessed the degree of plaque calcification (computed tomography), NPP1 and OPN localization (immunohistochemistry) and expression (RT-PCR) in a cohort of 45 patients undergoing carotid endatherectomy for significant stenosis of the internal carotid artery and in normal arteries (N= 50). We correlated NPP1 and OPN expression levels to the degree of plaque calcification, to pro-atherogenic factors and statin therapy. NPP1 was demonstrated in the base and in the shoulder of atherosclerotic plaques. Compared to normal arteries and non-calcified plaques, in calcified plaques NPP1 mRNA was decreased (P < 0.0001). OPN mRNA levels were up-regulated in carotid atheroma. NPP1 and OPN expression levels positively correlated with the degree of plaque calcification (R= 0.54, P= 0.00019 and R= 0.46, P= 0.017, respectively) and with risk factors of atherosclerosis. Expression of the calcification inhibitor NPP1 is down-regulated in calcified atherosclerotic plaques. Our correlation data point to a counter active mechanism, which in the end turns out to be insufficient to prevent further progression of calcification. PMID- 20015204 TI - Ferroportin 1 is expressed basolaterally in rat kidney proximal tubule cells and iron excess increases its membrane trafficking. AB - Ferroportin 1 (FPN1) is an iron export protein expressed in liver and duodenum, as well as in reticuloendothelial macrophages. Previously, we have shown that divalent metal transporter 1 (DMT1) is expressed in late endosomes and lysosomes of the kidney proximal tubule (PT), the nephron segment responsible for the majority of solute reabsorption. We suggested that following receptor mediated endocytosis of transferrin filtered by the glomerulus, DMT1 exports iron liberated from transferrin into the cytosol. FPN1 is also expressed in the kidney yet its role remains obscure. As a first step towards determining the role of renal FPN1, we localized FPN1 in the PT. FPN1 was found to be located in association with the basolateral PT membrane and within the cytosolic compartment. FPN1 was not expressed on the apical brush-border membrane of PT cells. These data support a role for FPN1 in vectorial export of iron out of PT cells. Furthermore, under conditions of iron loading of cultured PT cells, FPN1 was trafficked to the plasma membrane suggesting a coordinated cellular response to export excess iron and limit cellular iron concentrations. PMID- 20015203 TI - Small leucine-rich proteoglycans in atherosclerotic lesions: novel targets of chronic statin treatment? AB - Small leucine-rich proteoglycans (SLRPs), such as decorin and biglycan, regulate the assembly and turnover of collagenous matrix. The aim of the study was to analyse the effect of chronic rosuvastatin treatment on decorin, biglycan and the collagen matrix in ApoE-deficient mice. Twenty-week-old male ApoE-deficient mice received normal chow or 20 mg rosuvastatin/kg * day for 32 weeks. Subsequently, matrix composition was analysed by histochemistry and immunostaining at the aortic root and in innominate arteries of ApoE deficient mice as well as in human carotid endarterectomy specimens. Immunoblotting of proteoglycans was performed from aortic extracts of ApoE-deficient mice. Immunohistochemistry and immunoblotting revealed strongly increased decorin and biglycan deposition in atherosclerotic plaques at the aortic root and in innominate arteries. In contrast, versican and perlecan expression was not changed by rosuvastatin. Furthermore, matrix metalloproteinase 2 and gelatinolytic activity were decreased in response to rosuvastatin and a condensed collagen-rich matrix was formed. In carotid endarterectomy specimens of statin-treated patients increased decorin and biglycan accumulation was detected as well. Drug treatment did not change low density lipoprotein (LDL) plasma levels in ApoE-deficient mice and did not significantly affect lipid retention at the aortic root level as demonstrated by oil-red O staining and immunohistochemistry of LDL. Long-term treatment with rosuvastatin caused pronounced remodelling of atherosclerotic plaque matrix characterized specifically by enrichment with SLRPs and formation of a condensed collagen matrix. Therefore, decorin and biglycan might represent novel targets of statin treatment that contribute to a stable plaque phenotype. PMID- 20015205 TI - Evidence for the prevention of enthesitis in HLA-B27/hbeta(2)m transgenic rats treated with a monoclonal antibody against TNF-alpha. AB - Transgenic rats with high expression of HLA-B27 and human beta(2) -microglobulin (B27TR) develop a multisystem inflammatory disease resembling human inflammatory bowel disease (IBD) and spondyloarthropaties (SpA). Tumour necrosis factor alpha (TNF-alpha) has a crucial role in sustaining chronic inflammation in the gut and joints. The aim of this work was to evaluate whether TNF-alpha blockade could prevent or reduce the inflammation of peripheral joints in B27TR. A first group of 9-week-old B27TR received an anti-TNF-alpha monoclonal antibody (mAb) or an isotypic IgG2a,k up to the age of 18 weeks. An untreated group was monitored up to the age of 18 weeks and then randomly assigned to a 9-week treatment with anti TNF-alpha mAb or IgG2a,k. Each rat was monitored for clinical IBD and peripheral joint manifestations. After sacrifice the colon and hind paws were examined for macroscopical and microscopical pathological changes. Early TNF-alpha blockade prevented, and late treatment improved IBD signs in B27TR. Erythema, oedema, inflammatory infiltrate close to the tendons and enthesis, proliferating chondrocyte-like cells, signs of new endochondral bone ossification and bone erosion were observed in peripheral joints of four out of six IgG2a,k-treated B27TR, both at 18 and 27 weeks. Immunopositivity for phosphorylated Smad1/5/8 indicated that the process of joint remodelling was activated in B27TR. Some entheses showed chondroid nodules. Anti-TNF-alpha treatment reduced inflammation and preserved the enthesis organization in most animals. Occasional and transient erythema and oedema were still present in three of six of the late anti-TNF-alpha treated animals. Smad1/5/8 signalling was not inhibited by late anti-TNF-alpha treatment. In B27TR, articular involvement follows IBD onset and develops at entheses. Early TNF-alpha blockade prevents the onset of IBD and consequently the development of enthesitis in peripheral joints in the B27TR model of human SpA. PMID- 20015206 TI - Multiplex PCR detection of slowly-evolving trypanosomatids and neogregarines in bumblebees using broad-range primers. AB - AIMS: The aims of this study were to design universal markers for different protozoan parasites of Bombus spp. based on the phylogenetic position of two important bumblebee parasites Crithidia bombi and Apicystis bombi. METHODS AND RESULTS: Standard PCR and extraction techniques were used to amplify and sequence 18S rDNA. Phylogenetic analysis of the rDNA was performed in order to predict the parasite range of the primers. CONCLUSIONS: Crithidia bombi phylogenetically clusters with the trypanosomatids with slowly-evolving SSU-rRNA sequences (SE), while A. bombi is the closest sister group of Mattesia. A multiplex was designed containing an internal control and two broad-range primer pairs, detecting C. bombi and other SE trypanosomatids and also A. bombi and other neogregarines. SIGNIFICANCE AND IMPACT OF THE STUDY: Sequence data generated will further improve the current systematics of insect trypanosomatids and gregarines that remain troublesome. Broad-range markers for bumblebee parasites are necessary tools enabling the screening of commercially imported colonies and thus controlling their worldwide distribution and to discover related emerging parasites. PMID- 20015202 TI - Growth factors in ischemic stroke. AB - Data from pre-clinical and clinical studies provide evidence that colony stimulating factors (CSFs) and other growth factors (GFs) can improve stroke outcome by reducing stroke damage through their anti-apoptotic and anti inflammatory effects, and by promoting angiogenesis and neurogenesis. This review provides a critical and up-to-date literature review on CSF use in stroke. We searched for experimental and clinical studies on haemopoietic GFs such as granulocyte CSF, erythropoietin, granulocyte-macrophage colony-stimulating factor, stem cell factor (SCF), vascular endothelial GF, stromal cell-derived factor-1alpha and SCF in ischemic stroke. We also considered studies on insulin like growth factor-1 and neurotrophins. Despite promising results from animal models, the lack of data in human beings hampers efficacy assessments of GFs on stroke outcome. We provide a comprehensive and critical view of the present knowledge about GFs and stroke, and an overview of ongoing and future prospects. PMID- 20015207 TI - Dry thermal resistance of Bacillus anthracis (Sterne) spores and spores of other Bacillus species: implications for biological agent destruction via waste incineration. AB - AIMS: To obtain needed data on the dry thermal resistance of Bacillus anthracis spores and other Bacillus species for waste incinerator applications. METHODS AND RESULTS: Tests were conducted in a pilot-scale incinerator utilizing biological indicators comprised of spores of Geobacillus stearothermophilus, Bacillus atrophaeus and B. anthracis (Sterne) and embedded in building material bundles. Tests were also conducted in a dry heat oven to determine the destruction kinetics for the same species. In the pilot-scale incinerator tests, B. atrophaeus and G. stearothermophilus demonstrated similar thermal sensitivity, but B. anthracis (Sterne) was less thermally resistant than G. stearothermophilus. For the dry heat oven tests conducted at 175 degrees C, the D values were 0.4, 0.2 and 0.3 min for B. atrophaeus, B. anthracis (Sterne) and G. stearothermophilus, respectively. CONCLUSIONS: Bacillus anthracis (Sterne) possesses similar or less dry heat resistance compared to B. atrophaeus and G. stearothermophilus. SIGNIFICANCE AND IMPACT OF THE STUDY: Previous studies have demonstrated conditions under which bacterial spores may survive in an incinerator environment. The data from this study may assist in the selection of surrogates or indicator micro-organisms to ensure B. anthracis spores embedded in building materials are completely inactivated in an incinerator. PMID- 20015208 TI - Analysis of copper corrosion in compacted bentonite clay as a function of clay density and growth conditions for sulfate-reducing bacteria. AB - AIMS: To investigate the relationships between sulfate-reducing bacteria (SRB), growth conditions, bentonite densities and copper sulfide generation under circumstances relevant to underground, high-level radioactive waste repositories. METHODS AND RESULTS: Experiments took place 450 m underground, connected under in situ pressure to groundwater containing SRB. The microbial reduction of sulfate to sulfide and subsequent corrosion of copper test plates buried in compacted bentonite were analysed using radioactive sulfur (35SO4(2-)) as tracer. Mass distribution of copper sulfide on the plates indicated a diffusive process. The relationship between average diffusion coefficients (Ds) and tested density (rho) was linear. Ds (m2 s(-1))=-0.004xrho (kg m(-3))+8.2, decreasing by 0.2 Ds units per 50 kg m(-3) increase in density, from 1.2x10(-11) m2 s(-1) at 1750 kg m(-3) to 0.2x10(-11) m2 s(-1) at 2000 kg m(-3). CONCLUSIONS: It is possible that sulfide corrosion of waste canisters in future radioactive waste repositories depends mainly on sulfide concentration at the boundary between groundwater and the buffer, which in turn depends on SRB growth conditions (e.g., sulfate accessibility, carbon availability and electron donors) and geochemical parameters (e.g., presence of ferrous iron, which immobilizes sulfide). Maintaining high bentonite density is also important in mitigating canister corrosion. SIGNIFICANCE AND IMPACT OF THE STUDY: The sulfide diffusion coefficients can be used in safety calculations regarding waste canister corrosion. The work supports findings that microbial activity in compacted bentonite will be restricted. The study emphasizes the importance of growth conditions for sulfate reduction at the groundwater boundary of the bentonite buffer and linked sulfide production. PMID- 20015209 TI - The feasibility of developing a risk assessment for the impact of climate change on the emergence of Crimean-Congo haemorrhagic fever in livestock in Europe: a review. AB - Crimean-Congo haemorrhagic fever virus (CCHFV) is one of the most widespread of all medically important arboviruses with ticks of the Hyalomma spp. serving as the main vectors. Infection of livestock by CCHFV serves as a route of exposure to humans, as a reservoir of disease and as a route of importation. This study discusses the pathways and data requirements for a qualitative risk assessment for the emergence of CCHFV in livestock in Europe. A risk map approach is proposed based on layers that include the potential routes of release (e.g. by migrating birds carrying infected ticks) together with the main components for exposure, namely the distributions of the tick vectors, the small vertebrate host reservoirs and the livestock. A layer on landscape fragmentation serves as a surrogate for proximity of livestock to the tick cycle. Although the impact of climate change on the emergence of CCHF is not clear, comparing the distribution of risk factors in each layer currently with those predicted in the 2080s with climate change can be used to speculate how potential high-risk areas may shift. According to the risk pathway, transstadial and/or transovarial transmission in the tick vector are crucial for CCHFV spread. Vector competence and tick vector switching, however, remain critical factors for CCHFV colonization of new regions in Europe. The species of migratory bird is also an important consideration in the release assessment with greater abundance and biodiversity of ground-dwelling birds in southern Europe than in northern Europe. PMID- 20015210 TI - Testis response to low doses of cadmium in Wistar rats. AB - Although it is well known that cadmium (Cd) causes adverse effects on male rat reproductive organs, few studies have quantified alterations caused by its low doses. Quantification of these alterations, especially in the testis, was measured using morphometry. A single dose of cadmium chloride (1 or 1.2 mg/kg BW) was injected i.p. in adult rats, killed after 7 or 56 days. The lower dose caused slight alterations as measured by morphometrical analysis. The higher dose caused significant reduction in testis and epididymis weight, gonadossomatic index and length of seminiferous tubule (ST) after 7 and 56 days. Cadmium significantly reduced the ST diameter after 56 days. Decreased volume density of ST, after 7 and 56 days, was accompanied by an increase in interstitium volume density. The damage caused by the dose of 1.2 mg/kg can be clearly observed with light microscope. After 7 days, the tubule lumens were filled with degenerated germ cells and multinucleated spermatid aggregates. Vacuolization of the seminiferous epithelium was also observed. After 56 days, increased damage resulted in vacuolated ST, consisting only of Sertoli cells. Scanning electron microscopy examination of the testis showed that, in the group cadmium treated (1.2 mg/kg) and killed after 56 days, the interstitial tissue presents a compact and fibrous appearance with absence of fenestrae. The seminiferous epithelium height diminished and the absence of spermatozoa can be noted. The results show that a very small difference of Cd dose causes a sudden increase in testicular damage, apparently overpowering this tissue's natural defences. PMID- 20015211 TI - Aquagenic acrokeratoderma. AB - BACKGROUND: Aquagenic acrokeratoderma (AA) is a rare condition with female predilection that occurs after brief water exposure and disappears minutes to an hour after drying. The pathogenesis remains unclear. METHODS: Four Chinese patients with AA were reported and analyzed. RESULTS: There were 2 males and 2 females (age range: 14-33 years) who presented with a 2-week to more than 10-year history of small white papules coalescing into edematous plaques on the hands; lesions appeared within 5-10 minutes of water exposure, began to regress in 3-20 minutes and disappeared within 5 minutes to one hour after drying. Warm water provoked the lesions more rapidly than cold water. Lesions of a female patient could be triggered by detergent. In another female patient, lesions also involved the feet and were associated with palmoplantar erythema and hyperhidrosis. Biopsy from the lesion of one patient after water exposure revealed hyperkeratosis, mild hypergranulosis, and dilatated eccrine ducts. Biopsy from the lesion of another patient after drying showed normal epidermis and dermis. Two patients were treated with topical formalin 3% in alcohol, and two with 3% potassium aluminium sulfate solution with partial relief without any adverse effects. CONCLUSION: AA may occur in both males and females and may involve the feet. Warm water triggers the lesions more rapidly than cold. Topical formalin 3% in alcohol and 3% potassium aluminium sulfate solution may be optional therapy. PMID- 20015212 TI - The effect of seasonality, density and climate on the population dynamics of Montana deer mice, important reservoir hosts for Sin Nombre hantavirus. AB - 1. Since Sin Nombre virus was discovered in the U.S. in 1993, longitudinal studies of the rodent reservoir host, the deer mouse (Peromyscus maniculatus) have demonstrated a qualitative correlation among mouse population dynamics and risk of hantavirus pulmonary syndrome (HPS) in humans, indicating the importance of understanding deer mouse population dynamics for evaluating risk of HPS. 2. Using capture-mark-recapture statistical methods on a 15-year data set from Montana, we estimated deer mouse survival, maturation and recruitment rates and tested the relative importance of seasonality, population density and local climate in explaining temporal variation in deer mouse demography. 3. From these estimates, we designed a population model to simulate deer mouse population dynamics given climatic variables and compared the model to observed patterns. 4. Month, precipitation 5 months previously, temperature 5 months previously and to a lesser extent precipitation and temperature in the current month, were important in determining deer mouse survival. Month, the sum of precipitation over the last 4 months, and the sum of the temperature over the last 4 months were important in determining recruitment rates. Survival was more important in determining the growth rate of the population than recruitment. 5. While climatic drivers appear to have a complex influence on dynamics, our forecasts were good. Our quantitative model may allow public health officials to better predict increased human risk from basic climatic data. PMID- 20015213 TI - How do landscape composition and configuration, organic farming and fallow strips affect the diversity of bees, wasps and their parasitoids? AB - 1. Habitat destruction and increasing land use intensity result in habitat loss, fragmentation and degradation, and subsequently in the loss of species diversity. The fact that these factors are often highly confounded makes disentangling their effects extremely difficult, if not impossible, and their relative impact on species loss is mostly speculative. 2. In a two-year study, we analysed the relative importance of changed landscape composition (increased areas of cropped habitats), reduced habitat connectivity and reduced habitat quality on nest colonization of cavity-nesting bees, wasps and their parasitoids. We selected 23 pairs of conventional and organic wheat fields in the centre of landscape circles (500 m radius) differing in edge densities (landscape configuration) and % non crop habitats (landscape composition). Standardized trap nests were established in the field centres and in neighbouring permanent fallow strips (making a total of 92 nesting sites). 3. Factors at all three scales affected nest colonization. While bees were enhanced by high proportions of non-crop habitat in the landscape, wasps profited from high edge densities, supporting our hypothesis that wasps are enhanced by connecting corridors. Colonization of herbivore predating wasps was lower in field centres than in fallow strips for conventional sites, but not for organic sites, indicating a fallow-like connectivity value of organic fields. The relative importance of habitat type and farming system varied among functional groups suggesting that their perception of crop-non-crop boundaries or the availability of their food resources differed. 4. Local and landscape effects on parasitoids were mainly mediated by their hosts. Parasitism rates were marginally affected by local factors. A specialist parasitoid was more sensitive to high land use intensity than its host, whereas generalist parasitoids were less sensitive. 5. We conclude that the conversion of cropland into non-crop habitat may not be a sufficiently successful strategy to enhance wasps or other species that suffer more from isolation than from habitat loss. Interestingly, habitat connectivity appeared to be enhanced by both higher edge densities and by organic field management. Thus, we conclude that high proportions of conventionally managed and large crop fields threaten pollination and biological control services at a landscape scale. PMID- 20015214 TI - Breast lump detection: who is more accurate, patients or their GPs? AB - AIMS: A breast lump is the most commonly presenting complaint given by GPs when making a referral to the local breast unit. This does not necessarily mean that this was the breast symptom for which the woman consulted her GP. This study aimed to assess the accuracy of women and their GPs in detecting true breast lumps. MATERIALS AND METHODS: Between October 2008 and March 2009, the patients' symptoms, the signs elicited by the GP and the examination findings of the specialist breast clinician (BC) were recorded and compared for every consecutive new referral to the symptomatic breast clinic. A true lump was considered to present if it was detectable by the BC. RESULTS: Of the 282 referrals, 228 (81%) were for a lump. Women were 95% sensitive and 59% specific in detecting a true lump. GPs were 98% sensitive and 34% specific. Forty-six patients were referred by their GP with a lump but denied that one was present in themselves. Cancer was diagnosed in four of these women, three of whom suffered from dementia. DISCUSSION: Patients' and GPs' breast assessments are highly sensitive in detecting breast lumps. The specificity of GPs' assessments is low. The results reflect the high level of awareness that breast cancer usually presents as a lump and the priority given to not missing cancer. In the subgroup of cognitively unimpaired women referred with a lump, but who themselves deny that one is present, the patient will be correct and the GP incorrect in their assessment in 96% and 95% of patients respectively. CONCLUSION: If a woman consults her GP with breast symptoms, but has not found a lump, it is safe for her GP not to find one either. Such patients can be reassured and reviewed after a month with a view to referral, if symptoms persist. PMID- 20015216 TI - A novel point mutation in severe haemophilia A: a further proof of genotype phenotype correlation. PMID- 20015215 TI - Polymorphisms in genes involved in autoimmune disease and the risk of FVIII inhibitor development in Italian patients with haemophilia A. AB - One of the most severe and important complication in the treatment of patients with haemophilia A is the formation of neutralizing antibodies (FVIII inhibitors) that inhibit the clotting activity of substituted FVIII. Both genetic and environmental factors influence the susceptibility of patients to develop inhibitors. The objective of this study was to evaluate whether polymorphisms in different genes involved in the regulation of the immune system may confer susceptibility to inhibitor development in patients with HA. We analysed the distribution of polymorphisms in the CTLA4, PTPN22, IL10, TNFalpha, FOXP3 and IRF5 genes that have been reported to be associated with a number of autoimmune disease. In addition, we evaluated the distribution of IL10 haplotypes in haemophilic patients and healthy controls to assess whether specific polymorphisms in IL10 gene were associated to the risk of inhibitor development. We focused on a cohort of Italian unrelated haemophilic patients with and without a history of inhibitors. Genotyping was carried out with standard methods including RFLP, real time PCR and direct DNA sequencing. Our data show that, considering single nucleotide variations, genotype frequencies in patients with inhibitors were not significantly different from those observed in patients without inhibitors, suggesting a lack of association between these polymorphisms and the development of inhibitors. Moreover, no relationship was found between specific combinations of IL10 alleles and the antibody production. Previous contradictory association studies may depend on the different genetic background of the population examined. Further studies may contribute to a clearer understanding of this process. PMID- 20015217 TI - Characterization of the genetic basis of FXI deficiency in two Turkish patients. PMID- 20015218 TI - An open clinical study assessing the efficacy and safety of Factor IX Grifols, a high-purity Factor IX concentrate, in patients with severe haemophilia B. AB - SUMMARY: Factor IX Grifols is a new high-purity plasma-derived FIX concentrate with two specific pathogen elimination steps. Until this study was performed, there were no detailed reports with an adequate number of patients on the clinical evaluation of this product. To determine the efficacy and safety of Factor IX Grifols for replacement therapy in previously treated patients with severe haemophilia B, this open, multicentre and non-randomized study included 25 male subjects over the age of 12 with severe haemophilia B. Patients underwent prophylaxis and treatment of bleeding episodes with Factor IX Grifols for 1 year. The clinical efficacy and safety of this product were assessed. Forty percent of the patients were children and adolescents (12-17 years old). During the 12 months follow-up, 1 446 000 IU of Factor IX Grifols were administered in 961 infusions (range 12-83 infusions per patient): 31% for prophylaxis and 69% for bleeding episodes. Only five major bleeding events were reported in two patients. These haemorrhages were successfully treated with a mean of 2900 IU per bleed (range 1500-4000 IU), and 1-3 infusions per bleed. The average time elapsed from the first infusion to resolution of bleeding was 43 h (median). Overall, haemostasis was rated as excellent or good by the investigator in 96% of the infusions. No product-related adverse events were reported. Factor IX Grifols is an effective and safe Factor IX concentrate and can be considered as a first line option for replacement therapy in haemophilia B patients. PMID- 20015219 TI - Persistence of CCR5 usage among primary human immunodeficiency virus isolates of individuals receiving intermittent interleukin-2. AB - OBJECTIVE: To investigate the impact of intermittent interleukin-2 (IL-2) plus combination antiretroviral therapy (cART) on HIV-1 entry co-receptor use. METHODS: Primary HIV-1 isolates were obtained from 54 HIV-1-positive individuals at baseline and after 12 months using co-cultivation of peripheral blood mononuclear cells (PBMC) with activated PBMC of HIV-negative healthy donors. HIV 1 co-receptor use was determined on U87-CD4 cells. RESULTS: Fourteen out of the 21 (67%) IL-2-treated individuals harbouring a primary CCR5-dependent (R5) HIV-1 isolate at baseline confirmed an R5 virus isolation after 12 months in contrast to 3 out of 7 (43%) of those receiving cART only. After 12 months, only 1 R5X4 HIV-1 isolate was obtained from 21 cART+IL-2-treated individuals infected with an R5 virus at entry (5%) vs. 2/7 (29%) patients receiving cART alone, as confirmed by a 5-year follow-up on some individuals. CONCLUSIONS: Intermittent IL-2 administration plus cART may prevent evolution towards CXCR4 usage in individuals infected with R5 HIV-1. PMID- 20015220 TI - Attitudes towards new HIV biomedical prevention technologies among a cohort of HIV-negative gay men in Sydney, Australia. AB - OBJECTIVES: The aim of the study was to explore the awareness of rectal microbicides, the use of pre-exposure prophylaxis (PREP) and the willingness to participate in biomedical HIV prevention trials in a cohort of HIV-negative gay men. METHODS: In a community-based cohort study, HIV-negative homosexually active men in Sydney, Australia were questioned about awareness of rectal microbicides, use of PREP, and willingness to participate in trials of such products. Predictors of awareness and willingness to participate were analysed by logistic regression. Use of PREP was examined prospectively. RESULTS: Overall, 14% had heard of rectal microbicides. Older (P=0.05) and university-educated men (P=0.001) were more likely to have knowledge of rectal microbicides. Almost one quarter (24%) of men reported that they were likely/very likely to participate in rectal microbicide trials. Among those men with definite opinions on participation, awareness of rectal microbicides was significantly associated with unwillingness to participate [odds ratio (OR) 0.78, 95% confidence interval (CI) 0.65-0.93, P=0.007]. Willingness to participate in trials using antiretroviral drugs (ARVs) to prevent HIV infection was reported by 43% of men, and was higher among those who reported unprotected anal intercourse (UAI) with HIV-positive partners (OR 1.88, 95% CI 0.99-3.56). There was no evidence of current PREP use. CONCLUSIONS: This study demonstrates that Australian gay men have had little experience with PREP use and rectal microbicides. About half would be willing to consider participation in trials using ARVs to prevent HIV infection. Extensive community education and consultation would be required before PREP or rectal microbicides could be trialled in populations of gay Australian men. PMID- 20015221 TI - Ambiguous hydraulic heads and 14C activities in transient regional flow. AB - A regional flow and transport model is used to explore the implications of significant variability in Pleistocene and Holocene climates on hydraulic heads and (14)C activity. Simulations involve a 39 km slice of the Death Valley Flow System through Yucca Mountain toward the Amargosa Desert. The long-time scale over which infiltration has changed (tens-of-thousands of years) is matched by the large physical extent of the flow system (many tens-of-kilometers). Estimated paleo-infiltration rates were estimated using a juniper pollen percentage that extends from the last interglacial (LIG) period (approximately 120 kyrbp) to present. Flow and (14)C transport simulations show that groundwater flow changes markedly as a function of paleoclimate. At the last glacial maximum (LGM, 21 kyrbp), the recharge to the flow system was about an order-of-magnitude higher than present, and water table was more than 100 m higher. With large basin time constants, flow is complicated because hydraulic heads at a given location reflect conditions of the past, but at another location the flow may reflect present conditions. This complexity is also manifested by processes that depend on flow, for example (14)C transport. Without a model that accounts for the historical transients in recharge for at least the last 20,000 years, there is no simple way to deconvolve the (14)C dates to explain patterns of flow. PMID- 20015222 TI - Crude oil at the bemidji site: 25 years of monitoring, modeling, and understanding. AB - The fate of hydrocarbons in the subsurface near Bemidji, Minnesota, has been investigated by a multidisciplinary group of scientists for over a quarter century. Research at Bemidji has involved extensive investigations of multiphase flow and transport, volatilization, dissolution, geochemical interactions, microbial populations, and biodegradation with the goal of providing an improved understanding of the natural processes limiting the extent of hydrocarbon contamination. A considerable volume of oil remains in the subsurface today despite 30 years of natural attenuation and 5 years of pump-and-skim remediation. Studies at Bemidji were among the first to document the importance of anaerobic biodegradation processes for hydrocarbon removal and remediation by natural attenuation. Spatial variability of hydraulic properties was observed to influence subsurface oil and water flow, vapor diffusion, and the progression of biodegradation. Pore-scale capillary pressure-saturation hysteresis and the presence of fine-grained sediments impeded oil flow, causing entrapment and relatively large residual oil saturations. Hydrocarbon attenuation and plume extent was a function of groundwater flow, compound-specific volatilization, dissolution and biodegradation rates, and availability of electron acceptors. Simulation of hydrocarbon fate and transport affirmed concepts developed from field observations, and provided estimates of field-scale reaction rates and hydrocarbon mass balance. Long-term field studies at Bemidji have illustrated that the fate of hydrocarbons evolves with time, and a snap-shot study of a hydrocarbon plume may not provide information that is of relevance to the long term behavior of the plume during natural attenuation. PMID- 20015223 TI - Reliability of a method for evaluating porosity in denture base resins. AB - BACKGROUND: The method of porosity analysis by water absorption has been carried out by the storage of the specimens in pure water, but it does not exclude the potential plasticising effect of the water generating unreal values of porosity. OBJECTIVE: The present study evaluated the reliability of this method of porosity analysis in polymethylmethacrylate denture base resins by the determination of the most satisfactory solution for storage (S), where the plasticising effect was excluded. MATERIALS AND METHODS: Two specimen shapes (rectangular and maxillary denture base) and two denture base resins, water bath-polymerised (Classico) and microwave-polymerised (Acron MC) were used. Saturated anhydrous calcium chloride solutions (25%, 50%, 75%) and distilled water were used for specimen storage. Sorption isotherms were used to determine S. Porosity factor (PF) and diffusion coefficient (D) were calculated within S and for the groups stored in distilled water. anova and Tukey tests were performed to identify significant differences in PF results and Kruskal-Wallis test and Dunn multiple comparison post hoc test, for D results (alpha=0.05). RESULTS: For Acron MC denture base shape, FP results were 0.24% (S 50%) and 1.37% (distilled water); for rectangular shape FP was 0.35% (S 75%) and 0.19% (distilled water). For Classico denture base shape, FP results were 0.54% (S 75%) and 1.21% (distilled water); for rectangular shape FP was 0.7% (S 50%) and 1.32% (distilled water). FP results were similar in S and distilled water only for Acron MC rectangular shape (p>0.05). D results in distilled water were statistically higher than S for all groups. CONCLUSIONS: The results of the study suggest that an adequate solution for storing specimens must be used to measure porosity by water absorption, based on excluding the plasticising effect. PMID- 20015224 TI - Formation of 100S ribosomes in Staphylococcus aureus by the hibernation promoting factor homolog SaHPF. AB - In the stationary growth phase of Escherichia coli, the 70S ribosomes are dimerized by the ribosome modulation factor (RMF) and hibernation promoting factor (HPF) proteins to form 100S ribosomes, which lose translational activity. In this study we found 100S ribosomes in the gram-positive bacterium Staphylococcus aureus, which has an HPF homolog (named SaHPF) but no RMF homolog. Unlike in E. coli, 100S ribosomes exist in all growth phases of S. aureus, with the highest levels at the transition from the exponential phase to the stationary phase. To find the key factors involved in 100S formation, we analyzed proteins associated with crude ribosomes using radical-free and highly reducing 2-D PAGE and MALDI TOF/MS. Only the SaHPF levels changed in parallel with the changes in 100S levels. SaHPF bound preferentially to 70S components in 100S ribosomes, with a molar ratio of 1 : 1 relative to the 70S, but some SaHPF was also detected in free 70S ribosomes. High-salt washing of the crude ribosomes released SaHPF and dissociated the 100S ribosomes to their 70S components. When these 70S components were incubated with purified SaHPF in vitro, they re-associated to form 100S. These results suggest that SaHPF is a key protein involved in 100S ribosome formation in S. aureus. PMID- 20015225 TI - Indispensable function for embryogenesis, expression and regulation of the nonspecific form of the 5-aminolevulinate synthase gene in mouse. AB - The first step of heme biosynthesis in animals is catalyzed by 5-aminolevulinate synthase (ALAS), which controls heme supply in various tissues. To clarify the roles that the nonspecific isoform of ALAS (ALAS-N) plays in vivo, we prepared a green fluorescent protein (GFP) knock-in mouse line in which the Alas1 gene (encoding ALAS-N) is replaced with a gfp gene. We found that mice bearing a homozygous knock-in allele (Alas1(GFP/GFP)) were lethal by embryonic day 8.5, demonstrating that ALAS-N is essential for early embryogenesis. Fluorescence microscopic and flow cytometric analyses of heterozygous mouse (Alas1(+/GFP)) tissues showed that the Alas1 expression level differs substantially in tissues; Alas1 is highly expressed in testis Leydig cells, exocrine glands (including submandibular and parotid glands), endocrine glands (such as adrenal and thyroid glands) and hematopoietic lineage cells (including neutrophils and eosinophils). Quantitative analyses of GFP mRNA and ALAS-N mRNA in various tissues of Alas1(+/GFP) mice suggested that the destabilization of ALAS-N mRNA was not uniform in the various tissues. These results thus lay bare that elaborate control of the endogenous heme supply operates in various mouse tissues through regulation of the ALAS-N expression level and that this control is essential for heme homeostasis in animals. PMID- 20015226 TI - Effects of induced hyperthermia on pharmacokinetics of ropivacaine in rats. AB - Ropivacaine is a local anaesthetic used for epidural anaesthesia and postoperative pain relief. Hyperthermia is a very common sign of infection associated with variations in physiological parameters, which may influence drugs pharmacokinetics. The aim of this study was to determine the effects of induced hyperthermia on ropivacaine pharmacokinetics in rats. Two groups of six rats were given a single subcutaneous ropivacaine injection. Hyperthermia-induced animals were placed in a water bath to obtain a stable mean core temperature of 39.7 degrees C. After blood samples collection, ropivacaine serum concentrations and pharmacokinetic parameters were determined. Two other groups of six rats were sacrificed 30 min after ropivacaine injection to determine serum and tissues (brain and heart) concentrations. Our results (median +/- inter quartile range) reveal a significant increase of the total apparent clearance (0.0151 +/- 0.000800 L/min vs. 0.0134 +/- 0.00134 L/min), apparent volume of distribution (V(d)) (2.19 +/- 0.27 L vs. 1.57 +/- 0.73 L) and a significant decrease in exposure (488 +/- 50.6 mgxmin/L vs. 572 +/- 110 mgxmin/L) in induced-hyperthermia group. We observed a significant increase in brain ropivacaine concentration in hyperthermic rats (8.39 +/- 8.42 microg/g vs. 3.48 +/- 3.26 microg/g) and no significant difference between cardiac concentrations in the two groups (5.38 +/- 4.83 microg/g vs. 3.73 +/- 2.44 microg/g). Results suggest a higher tissular distribution of ropivacaine and an increase in blood-brain barrier permeability during hyperthermia. The hyperthermia-induced increase in V(d) could be responsible for an increase in cerebral ropivacaine toxicity. These experimental data provide a basis for future clinical investigations in relation to local anaesthetic use in hyperthermic patients. PMID- 20015227 TI - Antispasmodic effects of essential oil of Pterodon polygalaeflorus and its main constituent beta-caryophyllene on rat isolated ileum. AB - This study investigates the effects of essential oil of Pterodon polygalaeflorus (EOPP) and beta-caryophyllene (beta-CAR). EOPP and beta-CAR relaxed the basal tone of ileum smooth muscle in a concentration-dependent manner (IC(50) s = 394.35 +/- 62.12 and 68.65 +/- 9.51 MUg/mL respectively), an effect that was unaltered by hexamethonium, L-nitroarginine methyl ester or indomethacin. Both EOPP and beta-CAR evoked a concentration-dependent relaxation of ileum pre contracted with KCl with an IC(50) value of 107.78 +/- 10.47 and 17.35 +/- 0.75 MUg/mL, respectively. EOPP and beta-CAR inhibited the contractions induced by acetylcholine (ACh) and by KCl. In ileal preparations, the CaCl(2) -induced contractions were reduced by EOPP (300 MUg/mL) and beta-CAR (100 MUg/mL). Furthermore, CaCl(2) -induced contractions were also reduced by EOPP (300 MUg/mL) and beta-CAR (100 MUg/mL) in ileal preparations pretreated with ACh under Ca(2+) free condition and in the presence of verapamil. EOPP (100 and 300 MUg/mL) and beta-CAR (30 and 100 MUg/mL) reduced the ACh-induced contractions of isolated rat ileum under Ca(2+) -free conditions. In the presence of high KCl and Ca(2+) -free conditions, EOPP (300 MUg/mL) and beta-CAR (100 MUg/mL) reduced the contractions induced by barium. A similar effect was also observed with verapamil. It is concluded that (i) beta-CAR is an important constituent involved in the myorelaxant and antispasmodic effects induced by EOPP; (ii) the inhibitory effect on intestinal contractility is myogenic and seems mainly mediated through an intracellular mechanism. However, the ability of EOPP and beta-CAR to decrease Ca(2+) influx through cytoplasmic membrane could not be discounted. PMID- 20015228 TI - Antagonist-D2S-dopamine receptor interactions in intact recombinant Chinese hamster ovary cells [corrected]. AB - D(2)-type dopamine receptors are major recognition sites for antipsychotic drugs. There are two splice variants: D(2S) and D(2L) with an additional 29 amino acid sequence in the third intracellular loop. Only little comparative information is hitherto available about their pharmacological properties and none of these studies dealt with intact cell systems. This prompted us to investigate the binding properties of [(3)H]-raclopride, a hydrophilic benzamide, and [(3)H] spiperone, a highly hydrophobic butyrophenone, to intact CHO cells expressing recombinant human D(2L)-receptors. Presently, we have repeated and extended this experimental approach to the human D(2S)-receptors in the same cell system. Except for a slower dissociation of [(3)H]-spiperone from D(2S), the binding properties of these and other antagonists were not significantly different for both isoforms (P > 0.05). The very slow dissociation of the atypical antipsychotic clozapine was surprising in light of its low affinity. Two experiments pointed out the existence of non-competitive interactions between raclopride and spiperone for D(2S) as well as D(2L) (A. Packeu, J. P. De Backer & G. Vauquelin, in preparation). Alongside the different physicochemical properties of these ligands, this finding fits with a model wherein the hydrophilic raclopride approaches the D(2L)-receptor from the aqueous phase, while the hydrophobic spiperone approaches the receptor by lateral diffusion between the membrane lipids. These different modes of approach could imply the existence of topologically distinct ligand binding sites at D(2)-receptors. PMID- 20015229 TI - Diversity of pufM genes, involved in aerobic anoxygenic photosynthesis, in the bacterial communities associated with colonial ascidians. AB - Ascidians are invertebrate filter feeders widely distributed in benthic marine environments. A total of 14 different ascidian species were collected from the Western Mediterranean and their bacterial communities were analyzed by denaturing gradient gel electrophoresis (DGGE) of 16S rRNA gene. Results showed that ascidian tissues harbored Bacteria belonging to Gamma- and Alphaproteobacteria classes, some of them phylogenetically related to known aerobic anoxygenic phototrophs (AAPs), such as Roseobacter sp. In addition, hierarchical cluster analysis of DGGE patterns showed a large variability in the bacterial diversity among the different ascidians analyzed, which indicates that they would harbor different bacterial communities. Furthermore, pufM genes, involved in aerobic anoxygenic photosynthesis in marine and freshwater systems, were widely detected within the ascidians analyzed, because nine out of 14 species had pufM genes inside their tissues. The pufM gene was only detected in those specimens that inhabited shallow waters (<77 m of depth). Most pufM gene sequences were very closely related to that of uncultured marine bacteria. Thus, our results suggest that the association of ascidians with bacteria related to AAPs could be a general phenomenon and that ascidian-associated microbiota could use the light that penetrates through the tunic tissue as an energy source. PMID- 20015230 TI - Highlights from the first meeting of the Europe-Africa frontier research conference series infectious diseases: from basic to translational research. PMID- 20015231 TI - PCR-based plasmid typing in Enterococcus faecium strains reveals widely distributed pRE25-, pRUM-, pIP501- and pHTbeta-related replicons associated with glycopeptide resistance and stabilizing toxin-antitoxin systems. AB - A PCR-based typing scheme was applied to identify plasmids in an epidemiologically and geographically diverse strain collection of Enterococcus faecium (n=93). Replicon types of pRE25 (n=56), pRUM (n=41), pIP501 (n=17) and pHTbeta (n=14) were observed in 83% of the strains, while pS86, pCF10, pAM373, pMBB1 or pEF418 were not detected. Furthermore, 61% of the strains contained the axe-txe (n=42) or/and the omega-epsilon-zeta (n=18) plasmid stabilization loci. Sequence analyses divided the omega-epsilon-zeta operon into two distinct phylogenetic groups. The present typing scheme accounted for about 60% of the total number of plasmids detected by S1 nuclease analyses, which revealed zero to seven plasmids (10 kb to >200 kb) per isolate. Interestingly, strains belonging to the clinically important clonal complex 17 (CC17) yielded a significantly higher number of plasmids (3.1) and pRUM replicons (74%) than non-CC17 strains (2.2% and 35%, respectively). A prevalent genetic linkage between the pRUM replicon type and axe-txe was demonstrated by cohybridization analyses. The vanA resistance determinant was associated with all four replicon types, but we also confirmed the genetic linkage of vanA to unknown transferable replicons. PCR based replicon typing, linked to the detection of other important plasmid-encoded traits, seems to be a feasible tool for tracing disseminating resistance plasmids stably maintained in various environments. PMID- 20015232 TI - Neuroprotective effects of naturally occurring polyphenols on quinolinic acid induced excitotoxicity in human neurons. AB - Quinolinic acid (QUIN) excitotoxicity is mediated by elevated intracellular Ca(2+) levels, and nitric oxide-mediated oxidative stress, resulting in DNA damage, poly(ADP-ribose) polymerase (PARP) activation, NAD(+) depletion and cell death. We evaluated the effect of a series of polyphenolic compounds [i.e. epigallocatechin gallate (EPCG), catechin hydrate, curcumin, apigenin, naringenin and gallotannin] with antioxidant properties on QUIN-induced excitotoxicity on primary cultures of human neurons. We showed that the polyphenols, EPCG, catechin hydrate and curcumin can attenuate QUIN-induced excitotoxicity to a greater extent than apigenin, naringenin and gallotannin. Both EPCG and curcumin were able to attenuate QUIN-induced Ca(2+) influx and neuronal nitric oxide synthase (nNOS) activity to a greater extent compared with apigenin, naringenin and gallotannin. Although Ca(2+) influx was not attenuated by catechin hydrate, nNOS activity was reduced, probably through direct inhibition of the enzyme. All polyphenols reduced the oxidative effects of increased nitric oxide production, thereby reducing the formation of 3-nitrotyrosine and poly (ADP-ribose) polymerase activity and, hence, preventing NAD(+) depletion and cell death. In addition to the well-known antioxidant properties of these natural phytochemicals, the inhibitory effect of some of these compounds on specific excitotoxic processes, such as Ca(2+) influx, provides additional evidence for the beneficial health effects of polyphenols in excitable tissue, particularly within the central nervous system. PMID- 20015233 TI - Restriction point control of the mammalian cell cycle via the cyclin E/Cdk2:p27 complex. AB - Numerous top-down kinetic models have been constructed to describe the cell cycle. These models have typically been constructed, validated and analyzed using model species (molecular intermediates and proteins) and phenotypic observations, and therefore do not focus on the individual model processes (reaction steps). We have developed a method to: (a) quantify the importance of each of the reaction steps in a kinetic model for the positioning of a switch point [i.e. the restriction point (RP)]; (b) relate this control of reaction steps to their effects on molecular species, using sensitivity and co-control analysis; and thereby (c) go beyond a correlation towards a causal relationship between molecular species and effects. The method is generic and can be applied to responses of any type, but is most useful for the analysis of dynamic and emergent responses such as switch points in the cell cycle. The strength of the analysis is illustrated for an existing mammalian cell cycle model focusing on the RP [Novak B, Tyson J (2004) J Theor Biol230, 563-579]. The reactions in the model with the highest RP control were those involved in: (a) the interplay between retinoblastoma protein and E2F transcription factor; (b) those synthesizing the delayed response genes and cyclin D/Cdk4 in response to growth signals; (c) the E2F-dependent cyclin E/Cdk2 synthesis reaction; as well as (d) p27 formation reactions. Nine of the 23 intermediates were shown to have a good correlation between their concentration control and RP control. Sensitivity and co-control analysis indicated that the strongest control of the RP is mediated via the cyclin E/Cdk2:p27 complex concentration. Any perturbation of the RP could be related to a change in the concentration of this complex; apparent effects of other molecular species were indirect and always worked through cyclin E/Cdk2:p27, indicating a causal relationship between this complex and the positioning of the RP. PMID- 20015234 TI - Does mate limitation in self-incompatible species promote the evolution of selfing? The case of Leavenworthia alabamica. AB - Genetic diversity at the S-locus controlling self-incompatibility (SI) is often high because of negative frequency-dependent selection. In species with highly patchy spatial distributions, genetic drift can overwhelm balancing selection and cause stochastic loss of S-alleles. Natural selection may favor the breakdown of SI in populations with few S-alleles because low S-allele diversity constrains the seed production of self-incompatible plants. We estimated S-allele diversity, effective population sizes, and migration rates in Leavenworthia alabamica, a self-incompatible mustard species restricted to discrete habitat patches in rocky glades. Patterns of polymorphism were investigated at the S-locus and 15 neutral microsatellites in three large and three small populations with 100-fold variation in glade size. Populations on larger glades maintained more S-alleles, but all populations were estimated to harbor at least 20 S-alleles, and mate availabilities typically exceeded 0.80, which is consistent with little mate limitation in nature. Estimates of the effective size (N(e)) in each population ranged from 600 to 1600, and estimated rates of migration (m) ranged from 3 x 10( 4) to nearly 1 x 10(-3). According to theoretical models, there is limited opportunity for genetic drift to reduce S-allele diversity in populations with these attributes. Although pollinators or resources limit seed production in small glades, limited S-allele diversity does not appear to be a factor promoting the incipient breakdown of SI in populations of this species that were studied. PMID- 20015235 TI - Pronounced within-individual plasticity in sperm morphometry across social environments. AB - Sperm morphometry (i.e., size and shape) and function are important determinants of male reproductive success and are thought to be under stabilizing selection. However, recent studies suggest that sperm morphometry can be a phenotypically plastic trait, which can be adjusted to varying conditions. We tested whether different behavioral strategies in aggression between aggressive red and nonaggressive black males of the color polymorphic Gouldian finch (Erythrura gouldiae) can influence sperm morphometry. We show pronounced within-individual phenotypic plasticity in sperm morphometry of male Gouldian finches in three different social environments. Both red and black males placed in intermediate to high competitive environments (high frequency of red males) increased the relative length of their sperm midpiece. By contrast, red males placed in low to intermediate competitive environments (higher frequency of black males) increased the length of the sperm flagellum. Significant changes in stress and sex steroid hormone levels (in response to the competitive environment) appear to influence sperm traits in red but not in black males, suggesting that changes in hormonal levels are not solely responsible for the observed changes in sperm morphometry. These findings imply that males can adjust sperm morphometry across social environments. PMID- 20015236 TI - Sexual selection can increase the effect of random genetic drift--a quantitative genetic model of polymorphism in Oophaga pumilio, the strawberry poison-dart frog. AB - The variation in color pattern between populations of the poison-dart frog Oophaga pumilio across the Bocas del Toro archipelago in Panama is suggested to be due to sexual selection, as two other nonsexually selecting Dendrobatid species found in the same habitat and range do not exhibit this variation. We theoretically test this assertion using a quantitative genetic sexual selection model incorporating aposematic coloration and random drift. We find that sexual selection could cause the observed variation via a novel process we call "coupled drift." Within our model, for certain parameter values, sexual selection forces frog color to closely follow the evolution of female preference. Any between population variation in preference due to genetic drift is passed on to color. If female preference in O. pumilio is strongly affected by drift, whereas color in the nonsexually selecting Dendrobatid species is not, coupled drift will cause increased between-population phenotypic variation. However, with different parameter values, coupled drift will result in between-population variation in color being suppressed compared to its neutral value, or in little or no effect. We suggest that coupled drift is a novel theoretical process that could have a role linking sexual selection with speciation both in O. pumilio, and perhaps more generally. PMID- 20015237 TI - Influence of the indirect effects of guppies on life-history evolution in Rivulus hartii. AB - Early theories of life-history evolution predict that increased predation on young/small individuals selects for delayed maturation and decreased reproductive effort, but such theory only considers changes in mortality. Predators reduce prey abundance and increase food to survivors. Theory that incorporates such indirect effects yields different predictions. Trinidadian killifish, Rivulus hartii, inhabit communities with and without guppies. Guppies prey on young Rivulus and Rivulus densities decline and growth rates increase when guppies are present. Prior work showed that Rivulus phenotypes from communities with guppies matured earlier and had higher fecundity, consistent with theories that incorporate indirect effects. Here we examined the genetic basis of these differences by rearing 2nd generation, laboratory-born Rivulus from sites with and without guppies under two food levels that match natural differences in growth. Many locality x food interactions were significant, often reversing the relationship between communities. Such interactions imply that there are fitness trade-offs associated with adaptation to high or low resource environments. On high food, Rivulus from localities with guppies matured earlier, produced many small eggs, and exhibited increased reproductive investment; these differences reversed on low food. Our results suggest that indirect effects mold Rivulus evolution and thereby highlight connections between community processes and evolutionary change. PMID- 20015238 TI - The shape of contention: adaptation, history, and contingency in ungulate mandibles. AB - Mandibles and teeth of ungulates have been extensively studied to discern the functional significance of their design. Grazing ungulates have deeper mandibles, longer coronoid processes, flatter incisor arcades, and more hypsodont molars in comparison to browsers. If the functional significance of both mandible and teeth shapes is well-established, it remains uncertain to what extent mandible shapes are really adapted to grazing, meaning that they evolved either to serve their current biological function or just as a structural requirement to accommodate higher crowned molars. Here, we address this question by studying the contribution of phylogeny, hypsodonty, and body size to mandibular shape variation. The mandible shape appeared to be significantly influenced by hypsodonty but not by body size. Interestingly, hypsodonty-related changes influenced the tooth row in artiodactyls and perissodactyls significantly but in the opposite directions, which is ultimately related to their different digestive strategies. Yet, we obtained a strong phylogenetic effect in perissodactyls, suggesting that their mandible shape should be strongly inherited. The strength of this effect was not significant within artiodactyls (where hypsodonty explained much more variance in mandible shape). Digestive strategy is deemed to interplay with hypsodonty to produce different paths of adaptation to particular diets in ungulates. PMID- 20015239 TI - Size as a line of least resistance II: direct selection on size or correlated response due to constraints? AB - Evolutionary change in New World Monkey (NWM) skulls occurred primarily along the line of least resistance defined by size (including allometric) variation (g(max)). Although the direction of evolution was aligned with this axis, it was not clear whether this macroevolutionary pattern results from the conservation of within population genetic covariance patterns (long-term constraint) or long-term selection along a size dimension, or whether both, constraints and selection, were inextricably involved. Furthermore, G-matrix stability can also be a consequence of selection, which implies that both, constraints embodied in g(max) and evolutionary changes observed on the trait averages, would be influenced by selection. Here, we describe a combination of approaches that allows one to test whether any particular instance of size evolution is a correlated by-product due to constraints (g(max)) or is due to direct selection on size and apply it to NWM lineages as a case study. The approach is based on comparing the direction and amount of evolutionary change produced by two different simulated sets of net selection gradients (beta), a size (isometric and allometric size) and a nonsize set. Using this approach it is possible to distinguish between the two hypotheses (indirect size evolution due to constraints or direct selection on size), because although both may produce an evolutionary response aligned with g(max), the amount of change produced by random selection operating through the variance/covariance patterns (constraints hypothesis) will be much smaller than that produced by selection on size (selection hypothesis). Furthermore, the alignment of simulated evolutionary changes with g(max) when selection is not on size is not as tight as when selection is actually on size, allowing a statistical test of whether a particular observed case of evolution along the line of least resistance is the result of selection along it or not. Also, with matrix diagonalization (principal components [PC]) it is possible to calculate directly the net-selection gradient on size alone (first PC [PC1]) by dividing the amount of phenotypic difference between any two populations by the amount of variation in PC1, which allows one to benchmark whether selection was on size or not. PMID- 20015240 TI - Avian brood parasitism: information use and variation in egg-rejection behavior. AB - Hosts of avian brood parasites often vary in their response to parasitized clutches: they may eject one or several eggs, desert the nest, or accept all the eggs. Focusing on hosts exposed to single-egg parasitism by an evicting brood parasite, we construct an optimality model that includes all these behavioral options and use it to explore variation in rejection behavior. We particularly consider the influence of egg mimicry and external cues (observations of adult parasites near the nest) on optimal choice of rejection behavior. We find that several rejection responses will be present in a host population under a wide range of conditions. Ejection of multiple eggs tends to be adaptive when egg mimicry is fairly accurate, external cues provide reliable information of the risk of parasitism, and the expected success of renesting is low. If the perceived risk of parasitism is high, ejection of one or a few eggs may be the optimal rejection response even in cases in which hosts cannot discriminate between eggs. This may have consequences for the long-term outcome of the coevolutionary chase between hosts and parasites. We propose an alternative evolutionary pathway by which egg ejection may first arise as a defense against brood parasitism. PMID- 20015241 TI - Treatment response to bortezomib in multiple myeloma correlates with plasma hepatocyte growth factor concentration and bone marrow thrombospondin concentration. AB - Multiple myeloma (MM) is associated with increased rate of bone marrow angiogenesis. Increased concentration of hepatocyte growth factor (HGF) is associated with poor prognosis in patients treated with conventional chemotherapy or thalidomide. We have shown previously that decreased level of thrombospondin, an angiogenesis inhibitor, correlates with poor response to high-dose chemotherapy. The aim of our current study was to evaluate association between therapeutic response to bortezomib and thrombospondin and HGF levels. Peripheral blood plasma concentration of HGF and bone marrow plasma concentration of thrombospondin were measured in patients with MM prior to the initiation of bortezomib therapy. Overall, 58 patients were enrolled, 44/58 (76%) of them with relapsed disease. Treatment outcomes were analyzed for possible associations with pretreatment HGF and thrombospondin levels. Patients who achieved complete response had significantly higher pretreatment HGF levels and lower pretreatment thrombospondin levels than others. More than 70% of patients with low pretreatment HGF and high pretreatment thrombospondin concentrations achieved very good partial response or complete response, in contrast to only 20% of patients with high HGF and low thrombospondin levels. High pretreatment thrombospondin and low pretreatment HGF concentrations are associated with therapeutic response to bortezomib in patients with MM. PMID- 20015242 TI - Activity and safety of combination chemotherapy with methotrexate, ifosfamide, l asparaginase and dexamethasone (MILD) for refractory lymphoid malignancies: a pilot study. AB - Optimal salvage chemotherapy has not been established for lymphoid malignancy, which is refractory to the conventional cyclophosphamide, doxorubicin, vincristine, and prednisone regimen. To explore an effective regimen, we conducted a phase I pilot study of combination chemotherapy with methotrexate, ifosfamide, l-asparaginase and dexamethasone (MILD), which are unaffected by MDR1 encoded P-glycoprotein. A total of 18 patients with lethal lymphoid malignancy were enrolled over a 2-yr period. The median age was 63 yr. Eleven patients had T/NK-cell malignancies, six had B-cell malignancies, and one was diagnosed with a blastic plasmacytoid dendritic cell neoplasm. Patients aged >/=60 and <60 yr were planned to receive a set of starting doses of methotrexate and ifosfamide, which should induce myelosuppression. Eleven patients completed two courses of MILD therapy. Treatment-related death because of systemic mucormycosis was observed in one patient. Major treatment-related adverse events were grade 3 or more hematologic toxicities, which included lymphopenia corresponding to dose-limiting toxicity. The most common grade 3 non-hematologic toxicity was febrile neutropenia. Of the 14 evaluated patients, three achieved a complete response, and four showed a partial response. The overall response rate was 57%. It was very interesting that all of seven responders had T/NK-cell malignancies. MILD therapy was feasible and presented acceptable toxicity in patients with refractory or lethal lymphoid malignancies. The efficacy for T/NK-cell malignancies should be further evaluated. PMID- 20015244 TI - Circulating mononuclear cells nuclear factor-kappa B activity, plasma xanthine oxidase, and low grade inflammatory markers in adult patients with familial hypercholesterolaemia. AB - BACKGROUND: Few data are available on circulating mononuclear cells nuclear factor-kappa B (NF-kB) activity and plasma xanthine oxidase (XO) activity in heterozygous familial hypercholesterolaemia (FH). The goal of the study was to analyse circulating mononuclear cells NF-kB and plasma XO activities in FH patients. MATERIALS AND METHODS: Thirty FH index patients and 30 normoglycaemic normocholesterolaemic controls matched by age, gender, body mass index, abdominal circumference and homeostasis model assessment index were studied. Plasma XO and inflammatory markers were measured by standard methods. NF-kB was assayed in circulating mononuclear cells. RESULTS: Familial hypercholesterolaemia patients showed a significantly higher NF-kB (75.0 +/- 20.7 vs. 42.7 +/- 16.8 relative luminiscence units) and XO (0.44 +/- 0.13 vs. 0.32 +/- 0.09 mU mL(-1)) activities than controls. In addition, interleukin-1, interleukin-6, high sensitivity C reactive protein (hsCRP) and oxidized LDL (LDL-ox) were also significantly higher in FH patients. In the total group (FH and controls), XO was significantly associated with LDL-cholesterol (LDL-C), apolipoprotein B (apoB), NF-kB and hsCPR, and NF-kB activity was significantly associated with XO, hsCPR, LDL-ox, LDL-C and apoB plasma values. Using multiple regression analysis, XO was independently associated with hsCPR and NF-kB, and NF-kB activity in circulating mononuclear cells was independently associated with apoB and LDL-ox plasma values. CONCLUSION: Familial hypercholesterolaemia patients show increased activities of NF-kB and XO, and higher values of low grade inflammatory markers related to atherosclerosis. NF-kB activity was independently associated with apoB plasma values. These data could explain in part the high cardiovascular disease risk present in these patients. PMID- 20015245 TI - Selection of two lines of mice based on latency to onset of methionine sulfoximine seizures. AB - PURPOSE: In various animals methionine sulfoximine (MSO) induces tonic-clonic seizures resembling the most striking form of human epilepsies. The aim of the present study was to select two lines of mice based upon differences in their latency to MSO-dependent seizures, in order to characterize them. METHODS: Random crosses involving eight inbred mice strains were used to generate the starting population in which the first MSO challenge (75 mg/kg, i.p.) was performed. Two groups of 16 breeding pairs were established by mating mice having the shortest (MSO-Fast) and the longest (MSO-Slow) convulsion latencies. Mating and selection by latency to MSO (75 mg/kg, i.p.) was carried out over six generations. RESULTS: MSO-Fast mice presented a significantly shorter MSO latency, and were more susceptible to MSO than MSO-Slow ones were. Electroencephalography (EEG) alterations were observed during the preconvulsive period when MSO-Fast mice were submitted to 75 mg/kg of MSO, and MSO-Slow ones to 200 mg/kg. Using another convulsant, kainic acid, the latency to convulse of MSO-Fast mice was significantly shorter than that of the MSO-Slow ones, whereas no difference was observed in response to pentylenetetrazole (PTZ). MSO-dependent convulsions were completely antagonized by MK-801, and partially by valproic acid, suggesting a preferential involvement of glutamatergic pathways. DISCUSSION: The model that we have developed for MSO "sensitive" and "resistant" mice could allow for a better understanding of MSO mechanisms of epileptogenesis, and it may also constitute a useful approach for therapeutic actions of drugs. PMID- 20015246 TI - Patterns of analgesia for fractured neck of femur in Australian emergency departments. AB - OBJECTIVES: Fractured neck of femur is a common ED problem and poor pain management in this patient group can contribute significantly to their morbidity. The present study aims to describe current practices for managing pain in patients with fractured neck of femur in Australian ED and to identify real or potential barriers to providing analgesia. METHODS: Hospitals were invited to participate in a retrospective medical chart audit of patients with fractured neck of femur. At each site, 20 cases were randomly selected from the previous 12 months. Patient demographics, timing, type and method of analgesia in ED, use of pain scales and perceived barriers to analgesia were extracted from the medical chart. RESULTS: Data on 646 patients were collected from 36 hospitals in five Australian states. Most patients were elderly with a preponderance of women. One hundred and eighty-five (28.6%) patients had no record of analgesia administration in the ED and almost half of these had also not received prehospital analgesia. Intravenous morphine was the most frequently used analgesic and only 45 patients received a nerve block in the ED. The median time to first analgesia was 75 min after ED arrival. The most commonly reported barriers reported were cognitive impairment and language difficulties. CONCLUSIONS: Oligoanalgesia for fracture neck of femur in Australian ED is common and time to analgesia tends to be relatively slow. Regional techniques are infrequently used despite their recognized efficacy. Strategies for improving pain management in this cohort of ED patients need to be explored. PMID- 20015247 TI - Head injuries related to sports and recreation activities in school-age children and adolescents: data from a referral centre in Victoria, Australia. AB - OBJECTIVES: Head injuries (HI) in children are common and even mild HI can lead to ongoing cognitive and behavioural changes. We set out to determine the causes of sport-related HI in school-age children presenting to a large urban ED as a basis for future interventions. METHOD: Identification and medical record review of all sport-related HI in children aged 6-16 years at a tertiary children's hospital ED in Victoria, Australia, over a 1 year period. Information was collected on demographics, injury variables and radiology findings. HI were classified as mild, moderate and severe based on GCS and radiography reports. RESULTS: Over 12 months there were 406 HI in school-age children. Seventy per cent were male. A large number of HI (129; 33%) were related to sports. Of these, most were classified as mild and 13% were classified as moderate or severe. Among a range of sports, Australian Rules football was associated with more than 30% of all HI attributable to a sport and recreation cause. Equestrian activities were the main cause of moderate HI. CONCLUSION: The present study identified sports as a major cause of HI in the Victorian paediatric emergency setting with Australian Rules football the most commonly involved sport. Further prevention initiatives should consider targeting Australian Rules football and equestrian activities. PMID- 20015248 TI - Lack of effect on blood alcohol level of swabbing venepuncture sites with 70% isopropyl alcohol. AB - OBJECTIVE: It is standard practice to clean the skin using a non-alcohol containing swab before forensic blood alcohol sampling, because of the belief that the use of an alcohol-containing swab will contaminate the sample. The present study aimed to determine whether cleaning the skin with 70% isopropyl alcohol swabs, before venepuncture, alters measured blood alcohol level (BAL). METHODS: Volunteers aged >18 years had paired venous blood tests, which were drawn within 2 min of each other. One arm was swabbed with a 70% isopropyl alcohol swab and allowed to dry before venepuncture. The other was swabbed with saline, and these concurrent samples were used as controls. BAL was tested using the enzymatic method. Pathologists analysing the samples were blinded to the swabbing technique used. The mean differences and standard deviations of each of the paired samples were analysed using Student's t-test. RESULTS: Fifty-six paired venous blood samples were obtained from volunteers. Mean BAL in the isopropyl alcohol-swabbed group was 3.27 mg/dL with a standard deviation of 1.14 mg/dL. Mean BAL in the saline-swabbed group was 3.41 mg/dL with a standard deviation of 1.11 mg/dL. The mean difference was 0.14 mg/dL, with a standard error of 0.157. There was no statistically significant difference between the groups. CONCLUSIONS: The present study demonstrated that the use of 70% isopropyl alcohol swabs does not significantly affect BAL when used before venepuncture. This has implications that challenge current forensic blood alcohol sample acquisition. PMID- 20015249 TI - Extraordinarily widespread and fantastically complex: comparative biology of endosymbiotic bacterial and fungal mutualists of insects. AB - Endosymbiosis is a pervasive, powerful force in arthropod evolution. In the recent literature, bacterial symbionts of insects have been shown to function as reproductive manipulators, nutritional mutualists and as defenders of their hosts. Fungi, like bacteria, are also frequently associated with insects. Initial estimates suggest that insect-fungal endosymbionts are hyperdiverse, yet there has been comparatively little research investigating the roles that fungi play in their insect hosts. In many systems in which the bacterial symbionts are well characterized, the possible presence of fungi has been routinely ignored. Why has there been so little research on this important group of symbionts? Here, we explore the differences between fungal and bacterial endosymbiotic insect mutualists. We make predictions about why a bacterium or fungus might be found associated with an insect host given particular ecological, physiological, or evolutionary conditions. We also touch on the various hurdles for studying fungal vs. bacterial endosymbionts and potential future research directions. PMID- 20015250 TI - Cerebral palsy and assisted reproductive technologies: a case-control study. AB - AIM: To determine whether assisted reproductive technologies (ART) were more likely to be the method of conception in singletons with cerebral palsy (CP) than in those without CP. METHOD: Singletons with CP born between 1991 and 2004 were selected from the Victorian Cerebral Palsy Register and matched for birth year to two singletons randomly selected from the Victorian Perinatal Data Collection Unit. Data from both sources were linked to records from three ART centres. Conditional logistic regression was used to assess the association between CP and aspects of conception using ART. Multivariate models were adjusted for parity, previous miscarriages, sex, gestational age, birthweight, and weight for gestational age. RESULTS: We identified 1241 singletons with CP (males n=721 [58%], females n=420 [42/100]; motor type: spastic [87%; unilateral 37%; bilateral 63%], ataxic n=60 [5/100], dyskinetic n=46 [4/100], hypotonic n=29 [2/100], unknown n=25 [2/100]. Gross Motor Function Classification System levels were I n=363 [29/100], II n=297 [24/100], III n=137 [11/100], IV n=160 [13/100], V n=192 [15/100], and unknown n=92 [7/100]). Sixteen (1.3/100) of the children with CP and 25 (1.0/100) of 2482 children without CP were conceived using ART. There was no significant increase in the odds of children with CP being conceived using ART (adjusted odds ratio 1.19, 95% confidence interval (CI) 0.63, 2.24) nor in the odds of them being conceived by a subfertile couple without ART (adjusted odds ratio 2.7, 95% CI 0.87, 8.36). INTERPRETATION: Singleton conception using ART is not strongly associated with an increased risk of CP. PMID- 20015252 TI - Probable trigeminal autonomic cephalgia in a 3-month-old male infant. AB - To my knowledge trigeminal autonomic cephalgias (TACs) have not previously been reported in infancy. The diagnosis is dependent on an accurate history, including parents noting any physical signs at the time of the episode. Obtaining a clear history can be challenging when such symptoms occur in preverbal children. Similarly, physical signs, being transient, may have resolved by the time the parents take the child to a doctor. In addition, the investigations may also be normal. In such circumstances, taking a photograph during an episode can confirm the diagnosis. I describe a case of probable trigeminal autonomic cephalgia starting in a 3-month-old male infant who presented with screaming episodes associated with characteristic changes seen on his face. Investigations, including cranial magnetic resonance imaging, electroencephalography, and urinary catecholamines, were normal. The diagnosis was confirmed from a photograph taken by the parents at the time of the attack. As the condition is very rare in young children, there is little information available in the literature on using treatment for prophylaxis or for aborting acute episodes in this age group. PMID- 20015251 TI - Expectation of life and unexpected death in open spina bifida: a 40-year complete, non-selective, longitudinal cohort study. AB - AIM: The aim of our study was to investigate survival and causes of death in a complete cohort of open spina bifida at the mean age of 40 years. METHOD: We conducted a community-based, prospective study of 117 consecutive infants (50 males, 67 females) with open spina bifida whose backs were closed non-selectively within 48 hours of birth between 1963 and 1971 at Addenbrooke's Hospital, Cambridge, UK. Of those who survived to age 1 year, 89% (82 out of 92) had a cerebrospinal fluid shunt. In 2007, all survivors were surveyed by postal questionnaire backed up by telephone interview. Details of deaths were obtained from the Office for National Statistics, medical records, and autopsy reports, and Kaplan-Meier survival curves were constructed. RESULTS: One in three of the cohort (40/117) died before the age of 5 years. A further 26% (31/117) died during the next 35 years, over 10 times the national average. Half the deaths (16/31) after the age of 5 were sudden and unexpected. All occurred in the community and were followed by a coroner's autopsy. The most frequent causes of these unexpected deaths were epilepsy, pulmonary embolus, acute hydrocephalus, and acute renal sepsis. The prognosis for survival was strikingly poor in those with the most extensive neurological deficit. Only 17% (7/42) of those born with a high sensory level (above T11) survived to the mean age of 40 years, compared with 61% (23/38) of those with a low sensory level (below L3; p=0.001). INTERPRETATION: Doctors and care planners need to be aware that, contrary to previous suggestions, there is continuing high mortality throughout adult life in individuals with open spina bifida, and many deaths are unexpected. PMID- 20015253 TI - Developmental coordination disorder, sex, and activity deficit over time: a longitudinal analysis of participation trajectories in children with and without coordination difficulties. AB - AIM: Children with developmental coordination disorder (DCD) are known to participate in active play less than typically developing children. However, it is not known whether the activity deficit between children with and without DCD widens or diminishes over time. METHOD: Data were obtained from a large, prospective cohort study of children (baseline n=2278, total n=2470). Motor coordination was assessed for 2083 students using the short form of the Bruininks Oseretsky Test of Motor Proficiency. Participation in organized and free-play activities was assessed using a participation questionnaire on five occasions over 3 years. Mixed-effects modelling was used to examine differences in participation over time between children with probable DCD (pDCD, n=111, 46 males, 65 females) and their typically developing peers (n=1972, 1016 males, 956 females). The mean age for the whole sample was 9 years 11 months (SD 5 mo) at assessment 1, 10 years 5 months (SD 5 mo) at assessment 2, 10 years 11 months (SD 5 mo) at assessment 3, 11 years 4 months (SD 4 mo) at assessment 4, and 11 years 11 months (SD 4 mo) at assessment 5. RESULTS: Children with pDCD reported less participation in organized and free-play activities than their typically developing peers, and these differences persisted over time. Among males, the gap in participation in free-play activities between those with DCD and typically developing children diminished substantially over time; among females, it increased slightly. INTERPRETATION: DCD is associated with a persistent activity deficit in children. Its effect on participation appears to be particularly serious among females but may diminish with time among males. PMID- 20015254 TI - Can the Child Behavior Checklist be used to screen for motor impairment? AB - AIM: It has been suggested that one approach to identifying motor impairment in children is to use the Child Behavior Checklist (CBCL) as a screening tool. The current study examined the validity of the CBCL in identifying motor impairment. METHOD: A total of 398 children, 206 females and 192 males, aged from 3 years 9 months to 14 years 10 months were assessed on the McCarron Assessment of Neuromuscular Development to determine their motor ability. Parents completed the CBCL. RESULTS: The 'Clumsy' item on the CBCL was found to predict motor ability independent of the child's age, sex, and scores on other items of the CBCL. However, the sensitivity of the 'Clumsy' item in terms of identifying motor impairment was found to be a low 16.7% compared with specificity of 93.2%. The item 'Not liked' was also found to be a significant predictor of motor impairment. INTERPRETATION: Although the 'Clumsy' and 'Not liked' items were found to have a relationship with motor ability, they should not be relied upon to categorize children as motor impaired versus not impaired. It is possible that these items may be better indicators of motor impairment in children with developmental disorders such as attention-deficit-hyperactivity disorder, but clinical samples are needed to address this. PMID- 20015255 TI - Communities contain closely related species during ecosystem disturbance. AB - Predicting community and species responses to disturbance is complicated by incomplete knowledge about species traits. A phylogenetic framework should partially solve this problem, as trait similarity is generally correlated with species relatedness, closely related species should have similar sensitivities to disturbance. Disturbance should thus result in community assemblages of closely related species. We tested this hypothesis with 18 disturbed and 16 reference whole-lake, long-term zooplankton data sets. Regardless of disturbance type, communities generally contained more closely related species when disturbed. This effect was independent of species richness, evenness, and abundance. Communities already under stress (i.e., those in acidic lakes) changed most when disturbed. Species sensitivities to specific disturbances were phylogenetically conserved, were independent of body size, and could be predicted by the sensitivities of close relatives within the same community. Phylogenetic relatedness can effectively act as a proxy for missing trait information when predicting community and species responses to disturbance. PMID- 20015256 TI - Bronchial brushing cytology of a pulmonary fetal adenocarcinoma with a poorly differentiated component. PMID- 20015257 TI - Endobronchial ultrasound-guided transbronchial needle aspiration cytology: a state of the art review. AB - Endobronchial ultrasound-guided transbronchial needle aspiration (EBUS-TBNA) is a recently developed, accurate, safe and cost-effective technique that allows sampling of mediastinal lymph nodes and peribronchial lesions including pulmonary and mediastinal lesions. Its major indications are the nodal staging of non-small cell carcinomas of the lung, their restaging after chemotherapy and/or radiation, the diagnosis of sarcoidosis and of metastases from extrathoracic malignancies, and the diagnosis of mediastinal lymphadenopathy and masses of unknown aetiology. From our experience at the University of Minnesota and a comprehensive review of the literature, we discuss technical aspects of the procedure, its advantages and limitations in comparison with other methods of sampling mediastinal lymph nodes, focusing on the role of the cytopathologist in ensuring the effectiveness of the procedure. An algorithmic approach to the cytological diagnosis, starting with the determination of the adequacy of the sample, is also presented. PMID- 20015258 TI - Fine needle aspiration cytology of oncocytic carcinoma of the submandibular gland with pre-existing oncocytoma: a case report. PMID- 20015259 TI - Measuring and monitoring illegal use of natural resources. AB - Illegal use of natural resources is a threat to biodiversity globally, but research on illegal activities has methodological challenges. We examined 100 studies that empirically identify targeted resources, techniques used to procure resources illegally, locations of illegal activities, characteristics of typical violators, incentives driving illegal use of resources, magnitude of the problem of illegal use (e.g., quantities used), or frequency of illegal activity. We based our evaluation of the methods used in these studies on their ability to provide these empirical data, relative labor demands, training and technology requirements, and levels of uncontrollable bias. We evaluated eight different methods: law-enforcement records, indirect observation, self-reporting, direct observation, direct questioning, randomized response technique (a survey method designed to improve accuracy of responses to sensitive questions), forensics, and modeling. Different situations favored different methods, each with distinct advantages and limitations. Six context-specific factors-location of resource use (in situ vs. ex situ), budget, technology and training capacity, ease of detection of illegal activity, scope of illegal activity (limited vs. widespread), and researchers' willingness to accept bias in results-help narrow the choice of methods. Several methodological concerns applied to any study of illegal resource use: regular monitoring can detect trends; modeling can incorporate sampling error and data uncertainties; researchers must manage levels of bias that vary between methods; triangulation of results from multiple methods can improve accuracy. No method is a panacea, but a combination of techniques can help address the lack of data on illegal activity. Researchers empirically compared results from different methods in only four studies, and no one has compared more than two methods simultaneously. Conservation would benefit from more research focused on: methods comparisons that include cost effectiveness, time efficiency, and statistical rigor; unique applications of the eight techniques currently in use; and testing of new methods. PMID- 20015260 TI - Proposed definition of environmental damage illustrated by the cases of genetically modified crops and invasive species. AB - The introduction of non-native plant species and the release of genetically modified (GM) crops can induce environmental changes at gene to ecosystem levels. Regulatory frameworks such as the Convention on Biological Diversity or the EU Deliberate Release Directive aim to prevent environmental damage but do not define the term. Although ecologists and conservationists often refer to environmental effects of GM crops or invasive species as damage, most authors do not disclose their normative assumptions or explain why some environmental impacts are regarded as detrimental and others are not. Thus far, a concise definition of environmental damage is missing and is necessary for a transparent assessment of environmental effects or risks. Therefore, we suggest defining environmental damage as a significant adverse effect on a biotic or abiotic conservation resource (i.e., a biotic or abiotic natural resource that is protected by conservational or environmental legislation) that has an impact on the value of the conservation resource, the conservation resource as an ecosystem component, or the sustainable use of the conservation resource. This definition relies on three normative assumptions: only concrete effects on a conservation resource can be damages; only adverse effects that lead to a decrease in the value of the conservation resource can be damages; and only significant adverse effects constitute damage to a conservation resource. Applying this definition within the framework of environmental risk assessment requires further normative determinations, for example, selection of a threshold to distinguish between adverse and significant adverse effects and approaches for assessing the environmental value of conservation resources. Such determinations, however, are not part of the definition of environmental damage. Rather they are part of the definition's operationalization through assessment procedures, which must be grounded in a comprehensible definition of environmental damage. PMID- 20015261 TI - Genetic structure of herpetofauna on Halmahera Island, Indonesia: implications for Aketajawe-Lolobata National Park. AB - Genetic variation within species--a priority for biodiversity conservation--is influenced by natural selection, demography, and stochastic events such as genetic drift. We evaluated the role of these factors in 14 codistributed species of reptiles and amphibians on the Indonesian island of Halmahera by testing whether their molecular variation was correlated with geographic distance, ecology, riverine barriers, or Halmahera's paleoisland precursors. We found support for isolation by distance effects in four species. Two of these four were also significantly affected either by rivers or by ecology. A fifth species was significantly affected by ecology and a sixth was significantly affected by Halmahera's paleoislands. These findings--the most comprehensive survey of multispecies genetic variation on Halmahera to date--bode well for the efficacy of the recently established Aketajawe-Lolobata National Park in conserving a substantial component of vertebrate genetic variation on this island. Future success of conservation efforts will depend crucially, of course, on funding for and enforcement of conservation management of this park. PMID- 20015262 TI - Experimental test of postfire management in pine forests: impact of salvage logging versus partial cutting and nonintervention on bird-species assemblages. AB - There is an intense debate about the effects of postfire salvage logging versus nonintervention policies on regeneration of forest communities, but scant information from experimental studies is available. We manipulated a burned forest area on a Mediterranean mountain to experimentally analyze the effect of salvage logging on bird-species abundance, diversity, and assemblage composition. We used a randomized block design with three plots of approximately 25 ha each, established along an elevational gradient in a recently burned area in Sierra Nevada Natural and National Park (southeastern Spain). Three replicates of three treatments differing in postfire burned wood management were established per plot: salvage logging, nonintervention, and an intermediate degree of intervention (felling and lopping most of the trees but leaving all the biomass). Starting 1 year after the fire, we used point sampling to monitor bird abundance in each treatment for 2 consecutive years during the breeding and winter seasons (720 censuses total). Postfire burned-wood management altered species assemblages. Salvage logged areas had species typical of open- and early successional habitats. Bird species that inhabit forests were still present in the unsalvaged treatments even though trees were burned, but were almost absent in salvage-logged areas. Indeed, the main dispersers of mid- and late successional shrubs and trees, such as thrushes (Turdus spp.) and the European Jay (Garrulus glandarius) were almost restricted to unsalvaged treatments. Salvage logging might thus hamper the natural regeneration of the forest through its impact on assemblages of bird species. Moreover, salvage logging reduced species abundance by 50% and richness by 40%, approximately. The highest diversity at the landscape level (gamma diversity) resulted from a combination of all treatments. Salvage logging may be positive for bird conservation if combined in a mosaic with other, less-aggressive postfire management, but stand-wide management with harvest operations has undesirable conservation effects. PMID- 20015263 TI - Potential effects of United States-Mexico border hardening on ecological and human communities in the Malpai Borderlands. PMID- 20015264 TI - On target? Strategies and progress in the development of therapies for colorectal cancer targeted against WNT signalling. AB - Chemotherapy is an integral part of treatment for colorectal cancer (CRC), especially in the context of metastatic cancer. Targeted therapies in the form of monoclonal antibodies directed against the endothelial growth factor receptor or the vascular endothelial growth factor have met with success, and have demonstrated the advantages of molecularly targeted therapy in colorectal cancer. Nevertheless, CRC remains a major cause of death, which demonstrates the urgent need for improved treatment strategies. The pathway activated by the Wingless type mouse mammary tumour virus integration site (WNT) family members is constitutively active and promotes cancer growth in the majority of CRCs. As a result, there has been interest in developing therapeutics that circumvent it either by inhibiting WNT-mediated transcription or by inactivating the target genes. This review considers the current therapies approved for use in CRC and discusses the progress with therapies designed to target the WNT signalling pathway. PMID- 20015265 TI - Cleft closure for the treatment of pilonidal sinus disease. AB - AIM: Treatments for pilonidal sinus disease are numerous and prone to failure. In complex disease, the morbidity is high. In contrast with complex operations, the cleft closure procedure can be done simply and successfully with better cosmetic results. We present the results of a single-centre experience of this procedure. METHOD: One hundred and fifty patients had the operation; most were treated as a day case and many were operated under local anaesthetic. RESULTS: Primary healing occurred in 83 (60%) of 139 patients. Recurrences requiring surgery have been seen in 5.3%. The long-term cosmetic appearance has been noted to approach normality. CONCLUSION: Cleft closure is a simple and highly effective operation to treat recurrent or extensive pilonidal sinus disease. PMID- 20015266 TI - Iatrogenic perforation at colonic imaging. AB - AIM: Adequate colonic imaging is generally an invasive procedure with attendant risks, particularly perforation. Endoscopy, barium enema and computed tomographic colonography (CTC) are the main techniques for investigating patients with symptoms of, or screening for, colorectal cancer. The potential complications of these investigations have to be weighed against the benefits. This article reviews the literature on the incidence, presentation and management of iatrogenic colonic perforation at colonic imaging. METHOD: A literature review of relevant studies was undertaken using PubMed, Cochrane library and personal archives of references. Manual cross-referencing was performed, and relevant references from selected articles were reviewed. Studies reporting complications of endoscopy, barium enema and CT colonography were included in this review. RESULTS: Twenty-four studies were identified comprising 640,433 colonoscopies, with iatrogenic perforation recorded in 585 patients (0.06%). The reported perforation rate with double-contrast barium enema was between 0.02 and 0.24%. Serious complications with CTC were infrequent, though nine perforations were reported in a case series of 24,365 patients (0.036%) undergoing CTC. CONCLUSION: Perforation remains an infrequent and almost certainly under-reported, complication of all colonic imaging modalities. Risk awareness, early diagnosis and active management of iatrogenic perforation minimizes an adverse outcome. PMID- 20015267 TI - Impact of diagnostic procedures on patient management and hospitalization cost during the 2000 and 2005 enterovirus epidemics in Marseilles, France. AB - Enteroviruses are frequent aetiological agents of central nervous system infections in humans. In 2000 and 2005, two large outbreaks of Echovirus 30 (a member of species human enterovirus B) were observed in the University Hospitals of Marseilles (France). Between the two epidemics, the diagnostic protocols for enterovirus infection were modified, moving from viral cultures and classic RT PCR in 2000 to real-time RT-PCR in 2005. We compared some viral and epidemiological characteristics of the 2000 and 2005 outbreaks with special attention to diagnostic procedures and to the subsequent clinical management of patients. Despite similar virological and epidemiological characteristics during both outbreaks, our results show that real-time RT-PCR techniques used in 2005 noticeably shortened the period of time necessary to deliver diagnostic results and suggest that this was associated with a decrease in the duration of hospitalization for positive cases. In conclusion, this study suggests that the improvement of enterovirus diagnosis had a major financial impact on the management of the 2005 epidemic in Marseilles and may constitute an interesting example of how new diagnostic methods in microbiology can be self-financed through improvement in patient management. PMID- 20015268 TI - Short-term renal outcomes in African American and Caucasian donors following live kidney donation. AB - INTRODUCTION: Although African Americans (AA) are considered higher risk kidney donors than Caucasians, limited data are available regarding outcomes of AA donors. METHODS: We performed a single-center retrospective review of all kidney donors from 1993 to 2007 and evaluated race/ethnic differences in post-donation changes in renal function, incident proteinuria, and systolic blood pressure (SBP) using linear mixed models. RESULTS: A total of 336 kidney donors (63 AA, 263 Caucasian, 10 other) were evaluated. Before donation, AA had higher serum creatinine concentrations, estimated glomerular filtration rate (GFR) values, and SBP levels than Caucasians. No significant changes in SBP or renal function were observed between the two groups within the first year after donation, although results were limited by incomplete follow-up. CONCLUSION: AA had higher pre donation serum creatinine, GFR, and SBP values compared to Caucasians; however, the degree of change in renal function and blood pressure did not differ between groups following kidney donation. Although long-term studies are needed, our study suggests that AA and Caucasians experience similar short-term consequences after donation. The incomplete data available on donor outcomes in our center and in prior publications also indicates a global need to implement systems for structured follow-up of live kidney donors. PMID- 20015269 TI - Sharing kidneys across donor-service area boundaries with sensitized candidates can be influenced by HLA C. AB - The United Network for Organ Sharing (UNOS) implemented the virtual crossmatch system in UNet as a way to improve the likelihood of a negative crossmatch when kidneys are shared with HLA-sensitized candidates across donor service area (DSA) boundaries. The role of HLA C in that process is not universally appreciated. We recently experienced an unexpected positive flow T and B cell crossmatch for an imported, HLA zero-mismatched kidney because of donor-specific HLA C antibodies and transplanted it into the backup candidate. HLA C locus antigens were not typed by the OPO's laboratory that sent the kidney so the UNet virtual crossmatch could not "strike" our candidate from the UNOS match run. HLA C locus typing data of donors for kidneys our DSA imported from other DSAs revealed that C typing was not performed in 23% (14/60) and was discrepant with our molecular type for 10% (6/60) and was concordant in 67% (40/60) of cases. The rate of positive donor specific crossmatches was higher (83%) for HLA C discrepantly typed donors than for concordantly typed donors (44%). Sensitization for HLA C (42%) is less frequent than for A (80%) or B (83%) locus antigens but the immunogenicity of C locus antigens in patients who make C locus antibodies is equivalent in black and white patients. Finally, the transplant rate of imported kidneys into class I sensitized candidates was 24%, and C locus-sensitized candidates comprised 55% of those transplanted. PMID- 20015270 TI - Preservation of the donor pancreas for whole pancreas and islet transplantation. AB - Whole pancreas and islet cell transplantation are both reliant upon the procurement and preservation of a high quality donor pancreas for a successful outcome. In the climate of a reducing donor pool it is imperative that donor optimization, meticulous surgical retrieval and evidence based methods of preservation are practiced to ensure optimal graft quality. Moreover expanded criteria donors and novel methods of pancreas preservation have the potential to expand the number of usable grafts and increase the availability of these transplant modalities to suitable patients with diabetes. This article provides a review of the current literature surrounding donor management, surgical technique and the various technologies of organ preservation applicable to the donor pancreas. PMID- 20015271 TI - Anti-PR3 immune responses induce segmental and necrotizing glomerulonephritis. AB - Wegener's granulomatosis (WG) is a life-threatening autoimmune vasculitis that affects lungs, kidneys and other organs. A hallmark of WG is the presence of classic anti-neutrophil cytoplasmic antibodies (c-ANCA) against self-proteinase 3 (PR3). Little is known about the role of these antibodies and PR3-specific immune responses in disease development. In this study, we demonstrate that PR3-specific autoimmune responses are pathogenic in non-obese diabetic (NOD) mice with an impaired regulatory arm of the immune response. Immunization of autoimmunity prone NOD mice with rmPR3 (recombinant mouse PR3) in complete Freund's adjuvant (CFA) resulted in high levels of c-ANCA, without detectable disease development. However, when splenocytes from these immunized mice were transferred into immunodeficient NOD-severe combined immunodeficiency (SCID) mice, the recipient mice developed vasculitis and severe segmental and necrotizing glomerulonephritis. No disease developed in NOD-SCID mice that received splenocytes from the CFA-alone-immunized donors (controls), indicating that disease development depends upon PR3-specific immune responses. In contrast to the pathology observed in NOD-SCID mice, no disease was observed when splenocytes from rmPR3-immunized C57BL/6 mice were transferred into immunodeficient C57BL/6 RAG-1(-/-) mice, suggesting that complex and probably multi-genetic factors play a role in the regulation of disease development. PMID- 20015272 TI - Decrease of blood dendritic cells and increase of tissue-infiltrating dendritic cells are involved in the induction of Sjogren's syndrome but not in the maintenance. AB - We have demonstrated previously that, in primary Sjogren's syndrome (SS), immature myeloid dendritic cells (DCs) are decreased in blood and mature myeloid DCs are accumulated in salivary glands, suggesting recruitment of the myeloid DCs from blood to salivary glands. To verify whether this finding is universal in patients of not only primary SS but also secondary SS, in this study we analysed the blood DCs of secondary SS patients. We examined 24 secondary SS and 29 primary SS patients. A direct correlation between the decreased number of myeloid DCs and the duration of Sicca syndrome in primary and secondary SS was observed; namely, the reduction of myeloid DCs in blood was restored spontaneously with duration time of Sicca syndrome. We also examined the immunohistochemical staining of salivary glands of SS patients with monoclonal antibodies against fascin, CD11c and human leucocyte antigen DR (HLA-DR). Fascin(+) or CD11c(+)/HLA DR(+) mononuclear cells were present in the salivary glands of secondary SS patients, as in primary SS. However, fascin(+) mononuclear cells were barely detected in the salivary glands of a chronic phase of SS patients. We also found a negative correlation between the frequency of blood myeloid DCs and salivary gland-infiltrating DCs in secondary SS patients, as well as primary SS. Our results suggest that the reduction of blood myeloid DCs and preferential trafficking of myeloid DCs into salivary glands is a common event in the early stage of SS. Myeloid DCs may play essential roles in the pathogenesis of Sicca syndrome of SS by initiating T helper cell immune responses. PMID- 20015273 TI - Low numbers of CD8+ T lymphocytes in hereditary haemochromatosis are explained by a decrease of the most mature CD8+ effector memory T cells. AB - Low CD8(+) T lymphocyte numbers have long been described in hereditary haemochromatosis (HH). Recently, two conserved haplotypes localized near the microsatellite D6S105 at the major histocompatibility complex (MHC) class I region were described predicting the clinical expression of HH and the CD8(+) T lymphocyte numbers. The A-A-T haplotype was associated with a severe clinical expression of HH and low CD8(+) T lymphocyte numbers, while the G-G-G haplotype was associated with a milder clinical expression of HH and high CD8(+) T lymphocyte numbers. As CD8(+) T lymphocytes are a very heterogeneous population, in this study we analysed the CD8(+) subpopulations of naive, central memory (T(CM)) and effector memory (T(EM)), and further subsets of CD8(+) T(EM) cells in 47 HH patients and 68 controls. In addition, association studies were conducted between the conserved haplotypes and the CD8(+) T cell subpopulations in HH. Variations of the numbers of naive and central memory cells with age were similar between HH patients and controls. For T(EM) cells and the T(EM) CD27(-)CD28(-) subset no effect of age was observed in HH [R(2) = 0.001, not significant (n.s.) and R(2) = 0.01, n.s., respectively] contrasting with the increasing of these subpopulations with age in controls (R(2) = 0.09, P = 0.017 and R(2) = 0.22, P = 0.0005, respectively). Interestingly, patients homozygous for the A-A-T haplotype have lower numbers of CD8(+) T(EM) cells due especially to lower numbers of T(EM) CD27(-)CD28(-) (0.206 +/- 0.119 and 0.066 +/- 0.067 x 10(6) cells/ml, respectively) than patients carrying the G-G-G haplotype (0.358 +/- 0.195 and 0.246 +/- 0.202 x 10(6) cells/ml, respectively). This may suggest an inability of HH patients to differentiate the CD8(+) T cells into the most mature phenotype. PMID- 20015274 TI - Immunological and clinical profile of adult patients with selective immunoglobulin subclass deficiency: response to intravenous immunoglobulin therapy. AB - Selective immunoglobulin (Ig)G3 subclass deficiency in adults, especially its immunological profile, has not been described previously in detail. Therefore, a retrospective chart review was conducted to characterize the immune profile and clinical manifestations in adult patients with selective IgG3 deficiency. We reviewed the charts of 17 adult patients attending our subspeciality immunology clinic with a diagnosis of selective IgG3 deficiency. The following immunological test results were recorded: lymphocyte subsets, proliferative response to mitogens (phytohaemagglutinin, concanavalin A, pokeweed mitogen) and soluble antigens (mumps, Candida albicans, tetanus toxoid), specific antibody response to tetanus toxoid and pneumococcal antigens, neutrophil oxidative burst and natural killer cell cytotoxicity. In addition, we recorded information about the types of infections and other associated diseases, and response to intravenous immunoglobulin therapy (IVIG). In the majority of patients, lymphocyte subsets were normal. Proliferative responses to mitogens and antigens were decreased in 33% and 40% of patients, respectively. Specific antibody responses to tetanus were normal; however, responses to various pneumococcal serotypes were impaired in a subset of patients. Patients suffered from recurrent upper respiratory tract infections, which usually decreased in frequency and severity following treatment with IVIG. The majority of these patients also had concurrent atopic diseases in the form of allergic rhinitis or asthma. Selective IgG3 subclass deficiency should be considered in adults with recurrent upper respiratory tract infections with or without allergic rhinitis or asthma, who may have normal levels of total IgG. IVIG appears to be an effective therapy. PMID- 20015275 TI - From statistical significance to clinical relevance. PMID- 20015276 TI - A prostacyclin agonist with thromboxane inhibitory activity for airway allergic inflammation in mice. AB - BACKGROUND: ONO-1301 is a novel drug that acts as a prostacyclin agonist with thromboxane A(2) (TxA(2)) synthase inhibitory activity. We investigated the effect of ONO-1301 on development of airway allergic inflammation. METHODS: Mice sensitized and challenged to ovalbumin (OVA) received ONO-1301, OKY-046 (TxA(2) synthase inhibitor), beraprost, a prostacyclin receptor (IP) agonist, ONO-1301 plus CAY10449 (selective IP antagonist) or vehicle during the challenge period. Twenty-four hours after the OVA challenge, airway hyperresponsiveness (AHR) to methacholine was assessed and bronchoalveolar lavage was performed. Lung specimens were excised for goblet cell staining and analysis of lung dendritic cells (DCs). Bone marrow-derived dendritic cells (BMDCs) were generated, in the presence or absence of drugs, for analysis of DC function. RESULTS: Mice that received ONO-1301 showed significantly lower AHR, airway eosinophilia, T-helper type 2 cytokine levels, mucus production and lung DCs numbers than vehicle treated mice. These effects of ONO-1301 were mostly reversed by CAY10449. BMDCs treated with ONO-1301 alone showed lower DC functions, such as expression of costimulatory factors or stimulation to spleen T cells. CONCLUSIONS: These data suggest that ONO-1301 may suppress AHR and airway allergic inflammation through modulation of DCs, mainly mediated through the IP receptor. This agent may be effective as an anti-inflammatory drug in the treatment of asthma. PMID- 20015277 TI - Serum lactoferrin level as a serologic biomarker for allergic rhinitis. AB - BACKGROUND: Allergic rhinitis (AR) is a very common disease and a risk factor for allergic asthma. The discovery of new biomarkers for the early detection of AR would improve the clinical outcomes and reduce socio-economic burden. We sought to identify a novel serologic marker for detection of AR using a proteomic approach. METHODS: To identify the proteins involved in AR, comparative proteomics was applied using nasal lavage fluids (NLFs) taken before and after a nasal provocation test (NPT) with Dermatophagoides pteronyssinus (Dpt) in a subject with AR sensitized to Dpt. The clinical relevance of the identified proteins was evaluated by ELISA using NLFs and sera from the three study groups: Dpt-sensitive AR; asymptomatic Dpt-sensitive controls; and non-atopic healthy controls. The sensitivities and specificities of the candidate proteins for predicting AR were determined using receiver operating characteristic (ROC) curves. RESULTS: In proteomic analysis, lactoferrin expression was up-regulated after NPT. The validation study using ELISA showed a significantly lower serum lactoferrin level in the AR group than those of the other two groups (P<0.05, respectively). To discriminate between subjects with or without AR, the optimal serum cut-off level of lactoferrin was set at <307 ng/mL using the ROC curve. The sensitivity and specificity for predicting AR were 81.4% and 58%. When combined with serum Dpt-specific IgE level, the sensitivity and specificity for predicting AR were 76.7% and 79.2%. CONCLUSION: These results suggest that the serum lactoferrin level is associated with the phenotype of Dpt-sensitive AR, and in combination with the serum Dpt-specific IgE level, may be a potential serologic marker for early detection of AR. PMID- 20015278 TI - Factors influencing parenting in early childhood: a prospective longitudinal study focusing on change. AB - BACKGROUND: Parenting influences child outcomes but does not occur in a vacuum. It is influenced by socio-economic resources, parental health, and child characteristics. Our aim was to investigate the relative importance of these influences by exploring the relationship between changing parental health and socio-economic circumstances and changes in parenting. METHODS: Data collected from the Avon Longitudinal Study of Parents and Children were used to develop an eight-item parenting measure at 8 and 33 months. The measure covered warmth, support, rejection, and control and proved valid and reliable. Regression analysis examined changes in financial circumstance, housing tenure, marital status, social support, maternal health and depression, and their influence on parenting score. The final model controlled for maternal age, education, and baseline depression. RESULTS: Most mothers reported warm, supportive parenting at both times. Maternal depression was the only variable for which both positive and negative change was associated with changes in parenting score. Less depression was associated with better parenting scores and more depression with worse parenting scores. Improvements in social support and maternal general health were both associated with improved parenting scores, but for neither of these variables was deterioration associated with deterioration in parenting scores. Worsening financial circumstances predicted deterioration in parenting score, but improvements were not predictive of improvements in parenting. CONCLUSIONS: Programmes aiming to improve parental health and social support are likely to return greater dividends with regard to improving parenting than programmes that aim to reduce family poverty. PMID- 20015279 TI - Parents' descriptions and experiences of young children recently diagnosed with intellectual disability. AB - AIM: The aim of the present study was to explore the variation of parents' descriptions and experiences of their child that was recently identified to have an intellectual disability (ID). METHODS: The study applied interpretative phenomenological analysis and analysis of narrative style looking at content and form of parental narratives. Data was collected from nine fathers and eight mothers through semi-structured interviews within 6 months following diagnosis. RESULTS: Analysis revealed three factors indicating the parents' level of processing: (1) emotional expressions regarding the child - varying between limited (distanced or idealized) and balanced/affectionate; (2) experience of the disability - varying between preoccupation and acceptance; and (3) time orientation - varying in terms of flexibility and temporal focus. CONCLUSIONS: Although parents of children with ID describe negative emotions in relation to the child and the disability, most of these parents also describe positive emotions that seemed to balance the negative experiences. PMID- 20015280 TI - Newborn hearing screening programme outcomes in a research hospital from Turkey. AB - BACKGROUND: Universal neonatal hearing screening programmes are encouraged to define and manage hearing loss in early ages of life. The aim of this study is to introduce our 14-month three-step hearing screening programme results with 16 975 births in Turkey. METHODS: In healthy neonates, Transient Evoked Otoacoustic Emission (TEOAE) is served as the initial screening in the first day of life. In newborns that did not meet pass criteria TEOAE was repeated in 10-day period. If the second test was 'refer' again, the screening was completed with auditory brainstem response (ABR). Additionally, ABR was performed for the neonates with neonatal intensive care unit (NICU) requirement and at high audiologic risk. Neonates who failed the screening test with ABR were referred for further evaluation. RESULTS: A total of 15 323 newborns and 1652 NICU infants were tested. The screening coverage was 94.4%; 14 521 neonates (94.7%) passed the first screening step (TEOAE), while 802 (5.2%) neonate failed. In total, 322 (40.1%) of the neonates out of 802 was subjected to the second TEOAE after 10 days have failed and ABR was applied. From the neonates participated the third step (ABR) totalling 1974, 43 (2.17%) of neonates obtained a 'refer' response. Out of these 43 neonates, 17 neonates were (39.5%) NICU infants. From the 43 neonates, 38 cases (88.4%) were found to have hearing impairment. The false positive rate for first step screening with TEOAE was 4.9%; second step with TEOAE was 1.85% and for ABR was 0.25%. CONCLUSIONS: It is apparent that three step national hearing screening programme which has been applied for the latest years in Turkey is an accurate and non-invasive method to determine the congenital hearing loss. In the future, screening programmes could be rearranged with two steps as initial with TEOAE and retest with ABR and the coverage of the screening programme can be extended. PMID- 20015281 TI - Vitiligo is more than skin deep: a survey of members of the Vitiligo Society. AB - BACKGROUND: Vitiligo is a common, largely acquired skin disease of unknown aetiology, which causes a variable amount of skin and hair depigmentation in affected people. It affects over half a million people in the UK alone, and a massive 50 million people worldwide. AIM: To quantify the psychosocial burden of vitiligo in the UK, by estimating its effect on daily life. METHODS: All members (n = 1790) of the Vitiligo Society, a UK national patient support group, were sent a questionnaire. Survey questions included demographics, disease-related characteristics, effect of vitiligo on daily life, and psychosocial support measures for patients with vitiligo. Results. In total, 520 (29% of members) responded, of which 354 (68%) were women. Vitiligo affected the hands in 414 (80%) and the face in 394 (76%) of the respondents. Over half (56.6%) of respondents indicated that vitiligo moderately or severely affects their quality of life (QOL). Finding a cure or effective lasting treatment was the main priority for most affected respondents. Most respondents obtain information about their disease from nonmedical sources: 431 (83%) from the Vitiligo Society and 129 (25%) from the internet, compared with 61 (12.5%) from dermatologists. CONCLUSION: Vitiligo is a skin condition that moderately or severely affects the QOL of most patients. Although most patients look for a cure or long-lasting treatment, only 12.5% of respondents to our survey had obtained information from a dermatologist. Vitiligo is a common condition that affects more than the skin, and has profound psychosocial implications for affected patients. PMID- 20015282 TI - Cutaneous lupus erythematosus: clinical and immunological study of 308 patients stratified by gender. AB - BACKGROUND: Numerous studies involving systemic lupus erythematosus (SLE) have attempted to identify gender differences in patients with lupus erythematosus (LE). However, few reports on cutaneous lupus erythematous (CLE) have identified gender differences. AIM: To analyse and compare the prevalence and characteristics of the main clinical and immunological features of male and female patients with CLE. METHODS: The medical records of 103 (33.4%) male and 205 (66.6%) female patients with CLE who were treated as inpatients or outpatients between January 1985 and December 2000 were retrospectively studied. All patients were reviewed in detail stratified by a predefined protocol. RESULTS: Female patients had a higher prevalence of Raynaud's phenomenon (P < 0.01), chilblain lupus (P = 0.005), arthralgias (P = 0.001) and SLE (P < 0.01). Female patients were also more likely to have an increased erythrocyte sedimentation rate (P < 0.005), higher levels of antinuclear antibodies (P < 0.001) and decreased levels of C3 (P < 0.001), C4 (P < 0.01) and CH50 (P < 0.01). There was a higher prevalence of clinical and laboratory abnormalities in female patients who had both SLE and CLE than in male patients with both conditions. Conclusions. In our series, differences in the expression of CLE existed between male and female patients with respect to the type of lesions, systemic features, and immunological findings. PMID- 20015283 TI - Acquired perforating dermatosis associated with pregnancy. AB - Acquired perforating collagenosis (APC) is a rare condition in which altered collagen and elastin fibres undergo transepidermal elimination. The pathogenesis is unknown but it is usually associated with underlying systemic disease, most commonly diabetes mellitus and renal impairment. We describe a case of an acquired perforating disorder occurring in pregnancy. APC is a diagnosis that can easily be missed, especially in atypical cases such as ours, and clinicians and histopathologists need to be aware of this condition to avoid unnecessary patient morbidity. PMID- 20015285 TI - Unilateral generalized morphoea: could vibration be a stimulating factor? PMID- 20015284 TI - Facial corticosteroid addictive dermatitis in Guiyang City, China. AB - Facial corticosteroid addictive dermatitis (FCAD) has rapidly emerged as a health problem in China in recent years, because of the uncontrolled use of topical steroids. In total, 312 patients with FCAD from Guiyang, China were entered into a study. In this study, FCAD was classified into five types: dermatitis (n = 109; 34.9%), acne (n = 78; 25%), rosacea (n = 64; 20.5%), angiotelectasia (n = 30; 9.6%) and dermotrophia/hyperpigmentation (n = 31; 9.9%). Improper advertisement by manufacturers, availability of steroids without prescription, improper prescribing by doctors, and patient misuse of medication has resulted in the rapid emergence of FCAD in China. Public awareness and physician and patient education of the potential side-effects of topical corticosteroids should be improved. PMID- 20015286 TI - Two cases of palmar petechiae as a presentation of dermatitis herpetiformis. PMID- 20015287 TI - Lactobacillus rhamnosus GG induces tumor regression in mice bearing orthotopic bladder tumors. AB - The present gold standard for bladder cancer is Mycobacterium bovis, Bacillus Calmette Guerin (BCG) immunotherapy. But it has a non-responder rate of 30-50% and side effects are common. Lactobacillus casei strain Shirota has been reported to reduce the incidence of recurrence in bladder cancer patients and to cure tumor-bearing mice. Our aim was to determine if Lactobacillus rhamnosus GG (LGG) could be as efficacious as BCG in a murine model of bladder cancer. MB49 bladder cancer cells secreting human prostate-specific antigen were implanted orthotopically in female C57BL/6 mice and urinary prostate-specific antigen levels were used as a marker of tumor growth. Mice were treated with either live or lyophilized LGG given via intravesical instillation, or both oral and intravesical LGG given once a week for a period of 6 weeks starting at day 4 after tumor implantation. A comparison of LGG and BCG immunotherapy was also carried out. LGG therapy (live or lyophilized) significantly (P = 0.006) increased the number of cured mice. Cytokine arrays and immune cell recruitment analysis revealed differences between untreated, treated, cured, and tumor bearing mice. LGG therapy restored XCL1 levels to those in healthy bladders. LGG also recruited large numbers of neutrophils and macrophages to the tumor site. Intravesical LGG and BCG immunotherapy had cure rates of 89 and 77%, respectively, compared with 20% in untreated mice. LGG has the potential to replace BCG immunotherapy for the treatment of bladder cancer. PMID- 20015288 TI - Genomic landscape of meningiomas. AB - Meningiomas are one of the most common adult brain tumors. For most patients, surgical excision is curative. However, up to 20% recur. Currently, the molecular determinants predicting recurrence and malignant transformation are lacking. We performed retrospective global genetic and genomic analysis of 85 meningioma samples of various grades. Copy number alterations were assessed by 100K single nucleotide polymorphism arrays and correlated with gene expression, proliferation indices and clinical outcome. In addition to chromosome 22q loss, which was detected in the majority of clinical samples, chromosome 6q and 14q loss was significantly more common in recurrent tumors and was associated with anaplastic histology. Five "classes" of meningiomas were detected by gene expression analysis that correlated with copy number alterations, recurrent status and malignant histology. These classes more accurately identified recurrent tumors relative to Ki-67 index and extent of surgical resection, and highlight substantial expression heterogeneity between meningiomas. These data offer the most complete description of the genomic landscape of meningiomas, and provide broad genomic information that may be used to further stratify meningioma patients into prognostic risk groups. PMID- 20015289 TI - Vasopressin synthesis by the magnocellular neurons is different in the supraoptic nucleus and in the paraventricular nucleus in human and experimental septic shock. AB - Impaired arginine vasopressin (AVP) synthesis and release by the neurohypophyseal system, which includes the neurohypophysis and magnocellular neurons of the paraventricular and supraoptic nuclei, have been postulated in septic shock, but changes in this system have never been assessed in human septic shock, and only partially experimentally. We investigated AVP synthesis and release by the neurohypophyseal system in 9 patients who died from septic shock and 10 controls, and in 20 rats with fecal peritonitis-induced sepsis and 8 sham-operation controls. Ten rats died spontaneously from septic shock, and the others were sacrificed. In patients with septic shock, as in rats that died spontaneously following sepsis induction, AVP immunohistochemical expression was decreased in the neurohypophysis and supraoptic magnocellular neurons, whereas it was increased in the paraventricular magnocellular neurons. No significant change was observed in AVP messenger RiboNucleic Acid (mRNA) expression assessed by in situ hybridization in either paraventricular or supraoptic magnocellular cells. This study shows that both in human and experimental septic shock, AVP posttranscriptional synthesis and transport are differently modified in the magnocellular neurons of the supraoptic and paraventricular nuclei. This may account for the inappropriate AVP release in septic shock and suggests that distinct pathogenic mechanisms operate in these nuclei. PMID- 20015291 TI - Functional characterization of nonsynonymous single nucleotide polymorphisms in the human organic anion transporter 4 (hOAT4). AB - BACKGROUND AND PURPOSE: The human organic anion transporter (hOAT) family of transmembrane carrier proteins mediate the cellular flux of anionic substances, including certain hormones and anti-cancer drugs. hOAT4 is highly expressed at the apical membrane of the renal tubular cell and facilitates drug re-absorption in the kidney. In the present study, the impact of 10 nonsynonymous single nucleotide polymorphisms (SNPs) of hOAT4 on transport function in COS-7 cells was characterized. EXPERIMENTAL APPROACH: Transport uptake assay was used to assess the function of the variant transporters. Cell surface biotinylation and western blot analysis were used to investigate the expression characteristics of the transporter proteins. Comparative modelling was used to interpret the influence of nonsynonymous changes in terms of hOAT4 structure. KEY RESULTS: Four naturally occurring hOAT4 variants (L29P, R48Y, V155G and T392I) exhibited a significant loss of function. Substitution of leucine-29, which is a conserved residue in OATs, with a proline residue, impaired the synthesis or the apparent stability of the transporter and membrane insertion was disrupted in the R48Y variant. In the case of the V155G and T392I variants, impaired function was due to decreased affinity of the transporter for oestrone sulphate and impaired transporter substrate turnover respectively. The T392I variant was inhibited more extensively than the wild-type transporter by the cationic substrate tetraethyl ammonium. CONCLUSIONS AND IMPLICATIONS: Several naturally occurring SNPs encode variant hOAT4s that may impair the renal tubular re-absorption of important drug substrates. PMID- 20015290 TI - The accumulation and metabolism of zidovudine in 3T3-F442A pre-adipocytes. AB - BACKGROUND AND PURPOSE: Cultured pre-adipocytes accumulate and metabolize zidovudine (ZDV), but its mode of accumulation into these cells is unclear. We investigated the mode of accumulation of [(3)H]-ZDV, and the impact of changes in external pH and modulators of drug transporters on its accumulation and metabolism. EXPERIMENTAL APPROACH: The initial rate and steady-state accumulation of [(3)H]-ZDV were measured in 3T3-F442A cells. P-glycoprotein (P-gp) expression was detected by Western blotting. External pH was varied, and modulators of intracellular pH and drug transporters were used to study the mode of accumulation of ZDV. Phosphorylated ZDV metabolites were detected by high performance liquid chromatography. KEY RESULTS: Intracellular accumulation of ZDV was rapid, reaching equilibrium within 20 min; nigericin increased accumulation by 1.9-fold, but this did not alter the generation of ZDV mono-, di- and triphosphate. The accumulation and metabolism were pH dependent, being maximal at pH 7.4 and least at pH 5.1. Monensin, carbonyl cyanide p-trifluoromethoxy) phenyl hydrazone, brefeldin A, bafilomycin A1 and concanamycin A increased accumulation; 2-deoxyglucose, dipyridamole, thymidine and tetraphenylphosphonium inhibited accumulation. The accumulation was saturable; the derived K(d) and capacity of binding were 250 nmol per 10(6) cells and 265 nM respectively. 3T3-F442A cells express P-gp; inhibitors of P-gp (XR9576 and verapamil), P-gp/BCRP (GF120918), multidrug resistance protein (MRP) (MK571) and MRP/OATP (probenecid) increased the accumulation of ZDV. Saquinavir, ritonavir, amprenavir and lopinavir increased accumulation. CONCLUSIONS AND IMPLICATIONS: The accumulation of ZDV in 3T3-F442A cells was rapid, energy dependent, saturable and pH sensitive. Western blot analysis showed that 3T3-F442A cells express P-gp, and direct inhibition assays suggest that ZDV is a substrate of P-gp and MRP. PMID- 20015292 TI - Structure-function relationships of the N-terminus of receptor activity-modifying proteins. AB - The receptor activity-modifying proteins (RAMPs) are a family of three single transmembrane proteins that have been identified as accessory proteins to some G protein-coupled receptors (GPCRs). They can regulate their pharmacology, forward trafficking and recycling, depending on the GPCR. The best characterized receptor complexes formed by RAMPs and GPCRs are the calcitonin peptide family receptors. The association of RAMP1 with the calcitonin receptor-like receptor (CL) constitutes the calcitonin gene-related peptide receptor, whereas RAMP2 or 3 with CL generates adrenomedullin receptors. In this case, the RAMPs substantially alter the pharmacology and trafficking properties of this GPCR. Amylin receptor subtypes are formed from calcitonin receptor (CTR) interactions with RAMPs. Although the RAMPs themselves are not responsive to calcitonin peptide family ligands, there is clear evidence that they participate in ligand binding, although it is still unclear whether this is by directly participating in binding or through allosteric modulation of CL or CTR. A considerable amount of mutagenesis data have now been generated on RAMPs to try and identify the residues that play a role in ligand interactions, and to also identify which residues in RAMPs interact with CL and CTR. This review will focus on RAMP mutagenesis studies with CL, summarizing and discussing the available data in association with current RAMP models and structures. The data reveal key regions in RAMPs that are important for ligand binding and receptor interactions. PMID- 20015293 TI - Modulatory effects of neuropsychopharmaca on intracellular pH of hippocampal neurones in vitro. AB - BACKGROUND AND PURPOSE: The intracellular pH (pHi) of neurones is tightly regulated by, for example, membrane-bound acid-exchangers and loaders. Nevertheless, excessive bioelectric activity lowers steady-state pHi. In turn, even a moderate acidification can inhibit neuronal activity, a process believed to be part of a negative feedback loop controlling neuronal excitation. As moclobemide, an antidepressant, and also some antiepileptic drugs can reduce neuronal pHi in hippocampus slices in vitro, we screened a panel of currently used neuropsychopharmaca for comparable effects. EXPERIMENTAL APPROACH: BCECF-AM loaded hippocampal slices were superfused with 16 different neuroleptics, antidepressants and antiepileptics under bicarbonate-buffered conditions. Changes in steady-state pHi of CA3 neurones were measured fluorometrically. KEY RESULTS: The antipsychotics haloperidol, clozapine, ziprasidone, and the antidepressants amitriptyline, doxepin, trimipramine, citalopram, mirtazapine, as well as the anticonvulsive drug tiagabine reversibly reduced the steady-state pHi by up to 0.35 pH-units in concentrations of 5-50 microM. In contrast, venlafaxine, the anticonvulsants carbamazepine, clonazepam, gabapentin, lamotrigine, zonisamide, and the mood stabilizer lithium had no effect on neuronal pHi. CONCLUSION AND IMPLICATIONS: These data substantiate the view that clinically relevant concentrations of neuroleptics and antidepressants can mediate changes in neuronal pHi, which may contribute to their pharmacological mode of action. Effects on pHi should be taken into account when therapeutic or even harmful effects of these drugs are evaluated. PMID- 20015294 TI - Mechanism of the anti-obesity effects induced by a novel melanin-concentrating hormone 1-receptor antagonist in mice. AB - BACKGROUND AND PURPOSE: Melanin-concentrating hormone (MCH) is an orexigenic neuropeptide expressed in the lateral hypothalamus that is involved in feeding and body weight regulation. Intracerebroventricular infusion of a peptidic MCH1 receptor antagonist ameliorated obesity in murine models. Recently, small molecule MCH1 receptor antagonists have been developed and characterized for the treatment of obesity. However, little is known of the mechanism of the anti obesity effects of MCH1 receptor antagonists. EXPERIMENTAL APPROACH: To examine the mechanisms of action of the anti-obesity effect of MCH1 receptor antagonists more precisely, we conducted a pair-feeding study in mice with diet-induced obesity (DIO), chronically treated with an orally active and highly selective MCH1 receptor antagonist and examined changes in mRNA expression levels in liver, brown and white adipose tissues. We also assessed the acute effects of the MCH1 receptor antagonist in energy expenditure under thermoneutral conditions. KEY RESULTS: Treatment with the MCH1 receptor antagonist at 30 mg.kg(-1) for 1 month moderately suppressed feeding and significantly reduced body weight by 24%. In contrast, pair-feeding resulted in a smaller weight reduction of 10%. Treatment with the MCH1 receptor antagonist resulted in a higher body temperature compared with the pair-fed group. TaqMan and calorimetry data suggested that the MCH1 receptor antagonist also stimulated thermogenesis. CONCLUSIONS AND IMPLICATIONS: Our results indicate that an MCH1 receptor antagonist caused anti-obesity effects im mice by acting on both energy intake and energy expenditure. PMID- 20015295 TI - Advances in imaging of new targets for pharmacological intervention in stroke: real-time tracking of T-cells in the ischaemic brain. AB - BACKGROUND AND PURPOSE: T-cells may play a role in the evolution of ischaemic damage and repair, but the ability to image these cells in the living brain after a stroke has been limited. We aim to extend the technique of real-time in situ brain imaging of T-cells, previously shown in models of immunological diseases, to models of experimental stroke. EXPERIMENTAL APPROACH: Male C57BL6 mice (6-8 weeks) (n= 3) received a total of 2-5 x 10(6) carboxyfluorescein diacetate succinimidyl ester (CFSE)-labelled lymphocytes from donor C57BL6 mice via i.v. injection by adoptive transfer. Twenty-four hours later, recipient mice underwent permanent left distal middle cerebral artery occlusion (MCAO) by electrocoagulation or by sham surgery under isoflurane anaesthesia. Female hCD2 green fluorescent protein (GFP) transgenic mice that exhibit GFP-labelled T-cells underwent MCAO. At 24 or 48 h post-MCAO, a sagittal brain slice (1500 microm thick) containing cortical branches of the occluded middle cerebral artery (MCA) was dissected and used for multiphoton laser scanning microscopy (MPLSM). KEY RESULTS: Our results provide direct observations for the first time of dynamic T cell behaviour in living brain tissue in real time and herein proved the feasibility of MPLSM for ex vivo live imaging of immune response after experimental stroke. CONCLUSIONS AND IMPLICATIONS: It is hoped that these advances in the imaging of immune cells will provide information that can be harnessed to a therapeutic advantage. PMID- 20015296 TI - NS309 restores EDHF-type relaxation in mesenteric small arteries from type 2 diabetic ZDF rats. AB - BACKGROUND AND PURPOSE: The endothelium-derived hyperpolarizing factor (EDHF) type relaxation in mesenteric small arteries from 21 week old Zucker lean (ZL) and Zucker diabetic fatty (ZDF) rats was investigated using (6,7-dichloro-1H indole-2,3-dione 3-oxime) (NS309), a potent activator of small-conductance, calcium-activated potassium channel (SK(Ca)) and intermediate-conductance, calcium-activated potassium channel (IK(Ca)). EXPERIMENTAL APPROACH: In the presence of inhibitors of cyclooxygenase and nitric oxide synthase [indomethacin and N(omega)-nitro-L-arginine methyl ester (l-NAME), respectively], acetylcholine (ACh)-induced hyperpolarization and EDHF-type relaxation were investigated under isometric conditions in the wire myograph using 0.5 and 1 microM NS309 and/or selective blockers of SK(Ca) and IK(Ca) channels. Membrane potential was recorded with glass microelectrodes, and changes in the intracellular calcium concentration of endothelial cells were visualized by confocal microscopy. SK(Ca) expression was assessed by Western blotting. KEY RESULTS: In arteries from ZDF rats, ACh-induced relaxation and membrane hyperpolarization were attenuated and, compared with arteries from ZL rats, NS309 was less potent at causing relaxation. Incubation with 0.5 microM NS309 did not increase ACh-induced relaxation in arteries from ZDF rats significantly. However, 1 microM NS309 restored it (both in the absence and in the presence of indomethacin and l-NAME) without changing endothelial intracellular calcium concentration. The restored EDHF-type relaxation was more sensitive to TRAM-34 (1-[(2-chlorophenyl) diphenylmethyl]-1H pyrazole) (1 microM) than to apamin. Expression of the SK(Ca) channel was unaltered. CONCLUSIONS AND IMPLICATIONS: The attenuated EDHF-type relaxation in mesenteric small arteries from ZDF rats can be restored by NS309 without changes in the intracellular calcium concentration of endothelial cells. These results may have clinical implications for the treatment of endothelial dysfunction in overweight type 2 diabetic patients. PMID- 20015297 TI - Dopamine transporter down-regulation following repeated cocaine: implications for 3,4-methylenedioxymethamphetamine-induced acute effects and long-term neurotoxicity in mice. AB - BACKGROUND AND PURPOSE: 3,4-Methylenedioxymethamphetamine (MDMA) and cocaine are two widely abused psychostimulant drugs targeting the dopamine transporter (DAT). DAT availability regulates dopamine neurotransmission and uptake of MDMA-derived neurotoxic metabolites. We aimed to determine the effect of cocaine pre-exposure on the acute and long-term effects of MDMA in mice. EXPERIMENTAL APPROACH: Mice received a course of cocaine (20 mg*kg(-1), x2 for 3 days) followed by MDMA (20 mg*kg(-1), x2, 3 h apart). Locomotor activity, extracellular dopamine levels and dopaminergic neurotoxicity were determined. Furthermore, following the course of cocaine, DAT density in striatal plasma membrane and endosome fractions was measured. KEY RESULTS: Four days after the course of cocaine, challenge with MDMA attenuated the MDMA-induced striatal dopaminergic neurotoxicity. Co administration of the protein kinase C (PKC) inhibitor NPC 15437 prevented cocaine protection. At the same time, after the course of cocaine, DAT density was reduced in the plasma membrane and increased in the endosome fraction, and this effect was prevented by NPC 15437. The course of cocaine potentiated the MDMA-induced increase in extracellular dopamine and locomotor activity, following challenge 4 days later, compared with those pretreated with saline. CONCLUSIONS AND IMPLICATIONS: Repeated cocaine treatment followed by withdrawal protected against MDMA-induced dopaminergic neurotoxicity by internalizing DAT via a mechanism which may involve PKC. Furthermore, repeated cocaine followed by withdrawal induced behavioural and neurochemical sensitization to MDMA, measures which could be indicative of increased rewarding effects of MDMA. PMID- 20015299 TI - Mixed phenotype acute leukaemia with giant inclusions. PMID- 20015298 TI - Bimolecular fluorescence complementation: lighting up seven transmembrane domain receptor signalling networks. AB - There is increasing complexity in the organization of seven transmembrane domain (7TM) receptor signalling pathways, and in the ability of their ligands to modulate and direct this signalling. Underlying these events is a network of protein interactions between the 7TM receptors themselves and associated effectors, such as G proteins and beta-arrestins. Bimolecular fluorescence complementation, or BiFC, is a technique capable of detecting these protein protein events essential for 7TM receptor function. Fluorescent proteins, such as those from Aequorea victoria, are split into two non-fluorescent halves, which then tag the proteins under study. On association, these fragments refold and regenerate a mature fluorescent protein, producing a BiFC signal indicative of complex formation. Here, we review the experimental criteria for successful application of BiFC, considered in the context of 7TM receptor signalling events such as receptor dimerization, G protein and beta-arrestin signalling. The advantages and limitations of BiFC imaging are compared with alternative resonance energy transfer techniques. We show that the essential simplicity of the fluorescent BiFC measurement allows high-content and advanced imaging applications, and that it can probe more complex multi-protein interactions alone or in combination with resonance energy transfer. These capabilities suggest that BiFC techniques will become ever more useful in the analysis of ligand and 7TM receptor pharmacology at the molecular level of protein-protein interactions. PMID- 20015300 TI - Graft-versus-lymphoma effect. PMID- 20015301 TI - Bone marrow signal during PET imaging in Hodgkin lymphoma. PMID- 20015302 TI - Identifying practices and ideas to improve the implementation of maternal mortality reduction programmes: findings from five South Asian countries. AB - OBJECTIVE: The successful implementation of programmes to reduce maternal mortality is constrained by a 'know-do' gap: the disparity between what is known and the application of that knowledge in policy and practice. This study identified innovations, practices and ideas aimed to improve project and programme implementation. DESIGN: Cross-sectional. SETTING: Five South Asian countries: Afghanistan, Bangladesh, India, Nepal and Pakistan. SAMPLE: Sixteen projects and programmes, and 100 key informants. METHODS: In-depth review of documents, key informant interviews and focus-group discussions. MAIN OUTCOME MEASURES: Innovations and ideas to improve programme implementation, and their perceived effects. RESULTS: Delegation of duties to intermediate-level health workers, incentivisation of health workers, providing the means to overcome financial barriers for accessing care, quality improvements and knowledge transfer were examples of ideas put into practice to improve programme implementation. There was a perception that these improved service use and availability, but objective evidence was lacking. CONCLUSIONS: Some innovations, practices and ideas are supported by evidence of effect, and could be replicated, whereas others have not been formally evaluated. Testing of these innovations is required before more widespread adoption can be recommended, although experiences should be shared to narrow the 'know-do' gap, even though the evidence on beneficial effects remains unclear. PMID- 20015303 TI - The association between Mycoplasma genitalium and pelvic inflammatory disease after termination of pregnancy. AB - The prevalence and complications of Mycoplasma genitalium and Chlamydia trachomatis infections among women undergoing termination of pregnancy were studied in this nested case-control study at Malmo University Hospital, Sweden, during 2003 to 2007. The study comprised 2079 women presenting for termination of pregnancy. Forty-nine women with M. genitalium infection and 51 women with C. trachomatis infection, together with 168 negative control women, were evaluated. The prevalences of M. genitalium and C. trachomatis were 2.5% and 2.8%, respectively. The M. genitalium was strongly associated with post-termination pelvic inflammatory disease (odds ratio 6.29, 95% CI 1.56-25.2). The increased risk for pelvic inflammatory disease associated with M. genitalium infection after termination of pregnancy suggests a causal relationship. PMID- 20015304 TI - Birth outcomes in primiparous women who were raped as adults: a matched controlled study. AB - OBJECTIVE: To compare the duration of labour and the birth outcome in a group of primiparous women who had been raped after the age of 16, with a control group from the same birth cohort. DESIGN: Cohort study. SETTING: University Hospital of North Norway. SAMPLE: Fifty women raped as adults and 150 controls. METHODS: Data about birth outcomes in the first pregnancy were collected from the patient files and data concerning the assault were obtained in a subsequent pregnancy through consultations with the women who had been raped. Birth outcomes in the group of women who had been raped were compared with matched controls using a multivariable logistic regression model. MAIN OUTCOME MEASURES: Caesarean section, operative vaginal delivery and duration of labour. RESULTS: During their first delivery, the women who had been raped had an increased risk for caesarean section (adjusted odds ratio 15.7, 95% CI 5.0-49.1) and for assisted vaginal delivery (adjusted odds ratio 13.1, 95% CI 4.9-34.5) when compared with controls. The group of women who had been raped had a longer second stage of labour than the control group (120 versus 55 minutes, P < 0.01). They were more often single mothers, unemployed and smokers, and had a higher body mass index and more previous pregnancy terminations and miscarriages than the control group. CONCLUSIONS: The women who had been raped had a longer second stage of labour, and an increased risk of caesarean section and operative vaginal delivery compared with controls from the general birth cohort. These findings indicate that the consequences for delivery for women who had been raped as adults could be specific and may warrant particular attention. The birth experience of women who had been raped should also be illuminated in future studies. PMID- 20015305 TI - A 2-year observational study to determine the efficacy of a novel single incision sling procedure (Minitape) for female stress urinary incontinence. AB - OBJECTIVE: Assessment of the 2 year outcome of the Minitape procedure. DESIGN: A prospective observational study of women undergoing the Minitape procedure for urodynamic stress incontinence. Setting Two tertiary referral urogynaecology units in the north of England. POPULATION: Sixty women between November 2002 and March 2006. METHODS: Women attended a research clinic where they completed a standardised 1 hour pad test and were examined. Women were assessed preoperatively and postoperatively at 6 months, 1 year and 2 years. MAIN OUTCOME MEASURES: Success was determined by a negative 1 hour pad test (gain of <1 g) and no desire for further treatment for stress urinary incontinence. RESULTS: All procedures were completed with local anaesthesia, with no additional sedation in 82% of cases. Intra-operative and immediate postoperative complications were rare. Twelve women (20%) experienced mesh complications, half of which were considered to be serious adverse events requiring exit from the study. At 2 years following Minitape insertion, six women (10%) were defined as cured. CONCLUSIONS: Although feasible to perform, this procedure is associated with a substantially lower cure rate than that published previously for other procedures. Cure rates decline over the 2 year follow-up period, especially during the first 6 months. PMID- 20015306 TI - Different profiles of circulating angiogenic factors and adipocytokines between early- and late-onset pre-eclampsia. AB - OBJECTIVE: Circulating angiogenic factors have been shown to be important in the pathophysiology of pre-eclampsia. Blood levels of adipocytokines differ in pre eclampsia relative to controls and may also play an important role in disease pathogenesis. Differences in the circulating levels of these molecules were compared between matched normotensive controls and women with pre-eclampsia with onset before or at/after 32 weeks, and according to whether the women were of normal weight (18.5 < body mass index < 25) or overweight. DESIGN: A cross sectional study of 110 pregnant Japanese women who visited the Department of Obstetrics and Gynecology, Okayama University Hospital, Okayama, Japan. SETTING: Tertiary referral centre serving 2000 births. METHODS: Serum concentrations of soluble fms-like tyrosine kinase 1 (sFlt-1), placental growth factor (PlGF), soluble endoglin (sEng), adiponectin and leptin were measured in women with pre eclampsia and in normotensive controls matched for age, gestational week, parity and body mass index. Main outcome measures Serum levels of sFlt-1, PlGF, the sFlt 1/PlGF ratio, sEng, adiponectin and leptin. RESULTS: The sFlt-1/PlGF ratio in early-onset pre-eclampsia was significantly higher than that in late-onset pre eclampsia (112.0 +/- 30.2 versus 45.4 +/- 43.8, P = 0.037). There was a significant elevation of leptin in both subtypes relative to controls (early: 58.6 +/- 18.3 ng/ml versus 26.0 +/- 6.7 ng/ml, P = 0.001; late: 39.5 +/- 9.2 ng/ml versus 22.0 +/- 4.3 ng/ml, P = 0.005), but adiponectin was increased only in late-onset pre-eclampsia (36.5 +/- 13.4 microg/ml versus 12.0 +/- 4.3 microg/ml, P = 0.003). Significant differences in angiogenic factors and adiponectin were found between normal and overweight women only in late-onset pre eclampsia. CONCLUSIONS: These data suggest that there are different profiles of angiogenic factors and adipocytokines between women who develop early- and late onset pre-eclampsia. PMID- 20015307 TI - Management challenges of a live 12-week sub-hepatic intra-abdominal pregnancy. PMID- 20015308 TI - Preterm delivery and risk of subsequent cardiovascular morbidity and type-II diabetes in the mother. AB - OBJECTIVE: Preterm delivery has been shown to be associated with subsequent maternal cardiovascular morbidity. However, the impact of the severity and recurrence of preterm delivery on the risk of specific cardiovascular events and the metabolic syndrome in the mother, have not been investigated. DESIGN: National registry-based retrospective cohort study. SETTING: Women delivering in Denmark from 1978 to 2007. POPULATION: Women with a first singleton delivery (n = 782 287), and with a first and second singleton delivery (n = 536 419). METHODS: Cox proportional hazard models, with the gestational age stratified into four groups as primary exposure. We made adjustments for maternal age, year of delivery, hypertensive pregnancy disorders, fetal growth deviation, placental abruption and stillbirth. MAIN OUTCOME MEASURES: Subsequent maternal hypertension, ischaemic heart diseases, thromboembolism and type-II diabetes. RESULTS: After a first delivery at 32-36 completed weeks of gestation, the adjusted risk of subsequent type-II diabetes increased 1.89-fold (1.69-2.10) and the risk of thromboembolism increased 1.42-fold (1.24-1.62). Women having a preterm delivery in the first pregnancy and a term delivery in the second had a 1.58-fold (1.34-1.86) increased risk of type-II diabetes and a 1.18-fold (0.96 1.44) increased risk of thromboembolism. Women having two preterm deliveries had a 2.30-fold (1.71-3.10) increased risk of type-II diabetes and a 1.80-fold (1.29 2.50) increased risk of thromboembolism. CONCLUSIONS: Preterm delivery is independent of other pregnancy complications associated with subsequent maternal overt type-II diabetes and thromboembolism. The recurrence of preterm delivery will augment these risks. PMID- 20015309 TI - Fertility-sparing laparoscopic radical trachelectomy for young women with early stage cervical cancer. AB - OBJECTIVE: To report the results of laparoscopic radical trachelectomy (LRT) with respect to surgical, oncological and reproductive outcomes. DESIGN: Retrospective analysis. SETTING: University of Ulsan College of Medicine, Asan Medical Centre, Seoul, Korea. SAMPLE: Thirty-two consecutive patients who wish to preserve fertility with early stage cervical cancer. METHODS: Demographic, clinicopathologic, surgical, and follow-up data were obtained from patients' medical records. All patients agreed to telephone interviews to assess their menstrual and obstetrical outcomes. MAIN OUTCOME MEASURES: Surgical parameters, perioperative complication, disease-free survival, overall survival, return and pattern of menstruation and pregnancy rate. RESULTS: Thirty-two consecutive patients who wish to preserve fertility with early stage cervical cancer were offered LRT. In five patients, the planned LRT procedures were abandoned during the operations because of lymph node metastasis or parametrial involvement on frozen section. The mean age was 29 years (range, 22-37 years). The mean tumour size was 1.7 cm in diameter (range, 0.4-3.5 cm). The mean operating time was 290 min (range, 120-520 min) and the mean estimated blood loss was 332 ml (range, 50 1000 ml). Perioperative transfusion was required in six patients. There were no perioperative complications requiring further management. After a median follow up time of 31 months (range, 1-58 months), there was one recurrence and death from disease. Regular menstruation returned in 24 patients. Six patients attempted to conceive, and three succeeded. CONCLUSIONS: Laparoscopic radical trachelectomy may be a safe and useful alternative to radical hysterectomy for women with early cervical cancer who want to preserve their fertility. PMID- 20015310 TI - Urban-rural differences of gynaecological malignancies in Egypt (1999-2002). AB - OBJECTIVE: In previous studies, we have shown a three to four times higher urban incidence of breast cancer and estrogen receptor-positive breast cancers in the Gharbiah Province of Egypt. We investigated the urban-rural incidence differences of gynaecologic malignancies (uterine, ovarian and cervical cancers) to explore if they show the same trend that we found for breast cancer. DESIGN: Cancer registry-based incidence comparison. SETTING: Gharbiah population-based cancer registry (GPCR), Tanta, Egypt. SAMPLE: All patients with uterine, ovarian and cervical cancer in GPCR from 1999 to 2002. METHODS: We calculated uterine, ovarian and cervical cancer incidence from 1999 to 2002. For each of the three cancers, we calculated the overall and age-specific rates for the province as a whole, and by urban-rural status, as well as for the eight districts of the province. RESULTS: Incidence of all three cancer sites was higher in urban than in rural areas. Uterine cancer showed the highest urban-rural incidence rate ratio (IRR = 6.07, 95% CI = 4.17, 8.85). Uterine cancer also showed the highest urban incidence in the oldest age group (70+ age category, IRR = 14.39, 95% CI = 4.24, 48.87) and in developed districts (Tanta, IRR = 4.14, 95% CI = 0.41, 42.04). Incidence rates by groups of cancer sites showed an increasing gradient of urban incidence for cancers related to hormonal aetiology, mainly of the breast and uterus (IRR = 4.96, 95% CI = 2.86, 8.61). CONCLUSIONS: The higher urban incidence of uterine cancer, coupled with our previous findings of higher incidence of breast cancer and estrogen receptor positive breast cancer in urban areas in this region, may be suggestive of possible higher exposure to environmental estrogenic compounds, such as xenoestrogens, in urban areas. PMID- 20015311 TI - Floral evolution in the Annonaceae: hypotheses of homeotic mutations and functional convergence. AB - The recent publication of hypotheses explaining the homeotic control of floral organ identity together with the availability of increasingly comprehensive and well-resolved molecular phylogenies presents an ideal opportunity for reassessing current knowledge of floral diversity and evolution in the Annonaceae. This review summarizes currently available information on selected aspects of floral structure and function, including: changes in the number of perianth whorls and the number of perianth parts per whorl; the evolution of sympetaly; the diversity and evolution of pollination chambers (with a novel classification of seven main structural forms of floral chamber based on the different arrangement, size and shape of petals); the evolution of perianth glands; floral unisexuality and hypotheses explaining the unexpectedly high frequency of occurrence of androdioecy; the origin and possible function of inner and outer staminodes; the evolution of stamen connective diversity and theca septation; and the origin of 'true' syncarpy and functionally equivalent extragynoecial compita. In each case, current ideas on the origin, evolution and function are discussed. The information presented in this review enables two main conclusions to be drawn. The first is that changes in the homeotic control of floral organ identity may have had a profound impact on floral structure in several disparate lineages in the family. This is most obvious in Fenerivia, in which a centrifugal shift of floral organ identity has occurred, and in Dasymaschalon, in which a reverse (centripetal) shift has occurred. Other genera that have gained or lost entire perianth whorls are likely to have undergone similar homeotic changes. Attention is also drawn to the extensive functional convergence in Annonaceae flowers, with widespread homoplasy in many characters that have previously been emphasized in higher-level classifications. PMID- 20015312 TI - Spermatozoa of tapeworms (Platyhelminthes, Eucestoda): advances in ultrastructural and phylogenetic studies. AB - New data on spermiogenesis and the ultrastructure of spermatozoa of 'true' tapeworms (Eucestoda) are summarized. Since 2001, more than 50 species belonging to most orders of the Eucestoda have been studied or reinvestigated, particularly members of the Caryophyllidea, Spathebothriidea, Diphyllobothriidea, Bothriocephalidea, Trypanorhyncha, Tetraphyllidea, Proteocephalidea, and Cyclophyllidea. A new classification of spermatozoa of eucestodes into seven basic types is proposed and a key to their identification is given. For the first time, a phylogenetic tree inferred from spermatological characters is provided. New information obtained in the last decade has made it possible to fill numerous gaps in the character data matrix, enabling us to carry out a more reliable analysis of the evolution of ultrastructural characters of sperm and spermiogenesis in eucestodes. The tree is broadly congruent with those based on morphological and molecular data, indicating that convergent evolution of sperm characters in cestodes may not be as common as in other invertebrate taxa. The main gaps in the current knowledge of spermatological characters are mapped and topics for future research are outlined, with special emphasis on those characters that might provide additional information about the evolution of tapeworms and their spermatozoa. Future studies should be focused on representatives of those major groups (families and orders) in which molecular data indicate paraphyly or polyphyly (e.g. 'Tetraphyllidea' and Trypanorhyncha) and on those that have a key phylogenetic position among eucestodes (e.g. Diphyllidea, 'Tetraphyllidea', Lecanicephalidea, Nippotaeniidea). PMID- 20015313 TI - Resource pulses and mammalian dynamics: conceptual models for hummock grasslands and other Australian desert habitats. AB - Resources are produced in pulses in many terrestrial environments, and have important effects on the population dynamics and assemblage structure of animals that consume them. Resource-pulsing is particularly dramatic in Australian desert environments owing to marked spatial and temporal variability in rainfall, and thus primary productivity. Here, we first review how Australia's desert mammals respond to fluctuations in resource production, and evaluate the merits of three currently accepted models (the ecological refuge, predator refuge and fire-mosaic models) as explanations of the observed dynamics. We then integrate elements of these models into a novel state-and-transition model and apply it to well-studied small mammal assemblages that inhabit the vast hummock grassland, or spinifex, landscapes of the continental inland. The model has four states that are defined by differences in species composition and abundance, and eight transitions or processes that prompt shifts from one state to another. Using this model as a template, we construct three further models to explain mammalian dynamics in cracking soil habitats of the Lake Eyre Basin, gibber plains of the Channel Country, and the chenopod shrublands of arid southern Australia. As non equilibrium concepts that recognise the strongly intermittent nature of resource pulsing in arid Australia, state-and-transition models provide useful descriptors of both spatial and temporal patterns in mammal assemblages. The models should help managers to identify when and where to implement interventions to conserve native mammals, such as control burns, reduced grazing or predator management. The models also should improve understanding of the potential effects of future climate change on mammal assemblages in arid environments in general. We conclude by proposing several tests that could be used to refine the models and guide further research. PMID- 20015314 TI - Biological monitoring of non-thermal effects of mobile phone radiation: recent approaches and challenges. AB - This review describes recent developments in analysing the influence of radio frequency electromagnetic fields (RF-EMFs ) on biological systems by monitoring the cellular stress response as well as overall gene expression. Recent data on the initiation and modulation of the classical cellular stress response by RF EMFs, comprising expression of heat shock proteins and stimulation of stress activated protein kinases, are summarised and evaluated. Since isothermic RF-EMF exposure is assumed rather than proven there are clear limitations in using the stress response to describe non-thermal effects of RF-EMFs. In particular, further experiments are needed to characterise better the threshold of the thermal heat shock response and the homogeneity of the cellular response in the whole sample for each biological system used. Before then, it is proposed that the absence of the classical stress response can define isothermal experimental conditions and qualifies other biological effects of RF-EMFs detected under these conditions to be of non-thermal origin. To minimise the probability that by making this assumption valuable insights into the nature of biological effects of RF-EMFs could be lost, proteotoxic non-thermal RF-EMF effects should also be monitored by measuring activities of labile intracellular enzymes and/or levels of their metabolites before the threshold for the heat shock response is reached. In addition, non-thermal induction of the stress response via promoter elements distinct from the heat shock element (HSE) should be analysed using HSE-mutated heat shock promoter reporter constructs. Screening for non-thermal RF-EMF effects in the absence of a classical stress response should be performed by transcriptomics and proteomics. Recent approaches demonstrate that due to their high-throughput characteristics, these methods inherently generate false positive results and require statistical evaluation based on quantitative expression analysis from a sufficient number of independent experiments with identical parameters. In future approaches, positive results must be confirmed by independent quantitative methods and should also be evaluated in vivo to prove possible non-thermal effects of RF-EMFs on living beings. If successful, this strategy should contribute to identification of new underlying molecular mechanisms of interaction between RF-EMFs and living beings distinct from absorption of thermal energy. PMID- 20015315 TI - The ghosts of Gondwana and Laurasia in modern liverwort distributions. AB - Recent advances in phylogenetics and, in particular, molecular dating, indicate that transoceanic dispersal has played an important role in shaping plant and animal distributions, obscuring any effect of tectonic history. Taxonomic sampling in biogeographic studies is, however, systematically biased towards vertebrates and higher plants and the possibility remains that a much stronger signature of ancient vicariance might be evident among other organisms, particularly among basal land plants. Here, an explicit Bayesian model-based approach was used to investigate global-scale biogeographic patterns among liverwort genera and to determine whether the patterns identified are consistent with the expectations of vicariance or dispersal scenarios. The distribution of each genus was mapped onto the phylograms describing the floristic affinities among areas in order to define the synapomorphic transitions supporting the observed groupings. The probabilities of change in a branch were calculated by implementing the Markov model of BayesTraits. The consistent ambiguity in ancestral state reconstructions returned by the unconstrained, two-rate model indicated that the overall signal in the data was weak, leading us to test the performance of competing, explicit models. The analyses resolved clades of geographic areas that are mostly consistent with the kingdoms traditionally identified for plants and animals, but with strikingly lower rates of endemism. The major split observed in the phylograms is into almost entirely Laurasian and Gondwanan clades. Other patterns recovered by the analyses, including Wallace's line and the South Atlantic Disjunction, have also traditionally been interpreted in terms of vicariance. These observations contrast with the idea that, in spore dispersed organisms like bryophytes and pteridophytes, dispersal obscures evidence of vicariance. However, some discrepancies between the liverwort trees and expectations from a continental drift scenario were observed, such as the sister-group relationship of the Australian and New Zealand floras, which is supported by the co-occurrence of many genera, often endemic to these two areas. Together with an interpretation of the results within a phylogenetic context, our analyses suggest that patterns, which are at first sight consistent with an ancient vicariance hypothesis, may, in fact, conceal a complex mixture of relictual distributions and more recent, asymmetrical dispersal events. Our results provide a framework for testing specific evolutionary hypotheses concerning the extremely low levels of endemism in bryophytes and in particular, the significance of dispersal and cryptic diversification. PMID- 20015316 TI - Body size variation in insects: a macroecological perspective. AB - Body size is a key feature of organisms and varies continuously because of the effects of natural selection on the size-dependency of resource acquisition and mortality rates. This review provides a critical and synthetic overview of body size variation in insects from a predominantly macroecological (large-scale temporal and spatial) perspective. Because of the importance of understanding the proximate determinants of adult size, it commences with a brief summary of the physiological mechanisms underlying adult body size and its variation, based mostly on findings for the model species Drosophila melanogaster and Manduca sexta. Variation in nutrition and temperature have variable effects on critical weight, the interval to cessation of growth (or terminal growth period) and growth rates, so influencing final adult size. Ontogenetic and phylogenetic variation in size, compensatory growth, scaling at the intra- and interspecific levels, sexual size dimorphism, and body size optimisation are then reviewed in light of their influences on individual and species body size frequency distributions. Explicit attention is given to evolutionary trends, including gigantism, Cope's rule and the rates at which size change has taken place, and to temporal ecological trends such as variation in size with succession and size selectivity during the invasion process. Large-scale spatial variation in size at the intraspecific, interspecific and assemblage levels is considered, with special attention being given to the mechanisms proposed to underlie clinal variation in adult body size. Finally, areas particularly in need of additional research are identified. PMID- 20015317 TI - A framework for comparing pollinator performance: effectiveness and efficiency. AB - Measuring pollinator performance has become increasingly important with emerging needs for risk assessment in conservation and sustainable agriculture that require multi-year and multi-site comparisons across studies. However, comparing pollinator performance across studies is difficult because of the diversity of concepts and disparate methods in use. Our review of the literature shows many unresolved ambiguities. Two different assessment concepts predominate: the first estimates stigmatic pollen deposition and the underlying pollinator behaviour parameters, while the second estimates the pollinator's contribution to plant reproductive success, for example in terms of seed set. Both concepts include a number of parameters combined in diverse ways and named under a diversity of synonyms and homonyms. However, these concepts are overlapping because pollen deposition success is the most frequently used proxy for assessing the pollinator's contribution to plant reproductive success. We analyse the diverse concepts and methods in the context of a new proposed conceptual framework with a modular approach based on pollen deposition, visit frequency, and contribution to seed set relative to the plant's maximum female reproductive potential. A system of equations is proposed to optimize the balance between idealised theoretical concepts and practical operational methods. Our framework permits comparisons over a range of floral phenotypes, and spatial and temporal scales, because scaling up is based on the same fundamental unit of analysis, the single visit. PMID- 20015318 TI - Socioeconomic inequalities in coronary heart disease risk in older age: contribution of established and novel coronary risk factors. AB - BACKGROUND: Evidence on socioeconomic inequalities in coronary heart disease (CHD) and their pathways in the elderly is limited. Little is also known about the contributions that novel coronary risk factors (particularly inflammatory/hemostatic markers) make to socioeconomic inequalities in CHD. OBJECTIVES: To examine the extent of socioeconomic inequalities in CHD in older age, and the contributions (relative and absolute) of established and novel coronary risk factors. METHODS: A population-based cohort of 3761 British men aged 60-79 years was followed up for 6.5 years for CHD mortality and incidence (fatal and non-fatal). Social class was based on longest-held occupation recorded at 40-59 years. RESULTS: There was a graded relationship between social class and CHD incidence. The hazard ratio for CHD incidence comparing social class V (unskilled workers) with social class I (professionals) was 2.70 [95% confidence interval (CI) 1.37-5.35; P-value for trend = 0.008]. This was reduced to 2.14 (95% CI 1.06-4.33; P-value for trend = 0.11) after adjustment for behavioral factors (cigarette smoking, physical activity, body mass index, and alcohol consumption), which explained 38% of the relative risk gradient (41% of absolute risk). Additional adjustment for inflammatory markers (C-reactive protein, interleukin-6, and von Willebrand factor) explained 55% of the relative risk gradient (59% of absolute risk). Blood pressure and lipids made little difference to these estimates; results were similar for CHD mortality. CONCLUSIONS: Socioeconomic inequalities in CHD persist in the elderly and are at least partly explained by behavioral risk factors; novel (inflammatory) coronary risk markers made some further contribution. Reducing inequalities in behavioral factors (especially cigarette smoking) could reduce these social inequalities by at least one-third. PMID- 20015319 TI - Evaluation of anti-activated protein C antibody development in patients with severe sepsis from four clinical studies with drotrecogin alpha (activated). AB - BACKGROUND: Drotrecogin alpha (activated) (DAA) is a recombinant human activated protein C (APC), which is an antithrombotic protein. OBJECTIVES: To evaluate the development of anti-APC antibodies in severe sepsis patients in DAA clinical studies. PATIENTS AND METHODS: Serum and plasma samples were collected in placebo controlled studies (PROWESS, ADDRESS) and studies where all patients were DAA treated (ENHANCE, XPRESS). An enzyme-linked immunosorbent assay detecting anti APC IgA/IgG/IgM antibodies was used. IgG isolated from plasma of positive samples was tested for neutralizing activity against DAA-induced prolongation of activated partial thromboplastin time. RESULTS: The proportions of patients with negative baseline but positive postbaseline anti-APC antibodies were 1.5% (27/1855) and 1.6% (24/1493) in the DAA and placebo cohorts, respectively (P = 0.72 for the difference). Of the 27 DAA and 24 placebo patients with positive anti-APC antibodies, all but one (DAA) were alive at day 28, and all but seven (four DAA and three placebo) were alive at hospital discharge, including eight (five DAA and three placebo) patients who tested positive for anti-APC neutralizing antibodies. Two of the 51 patients who tested positive for the development of anti-APC antibodies experienced a thrombotic event (one DAA, one placebo). In ADDRESS, no anti-APC antibody was detected in the six DAA-treated patients who had received a previous course of DAA therapy. CONCLUSIONS: The proportion of patients with anti-APC antibodies was low and was similar between DAA-treated and placebo-treated patients. No relationship between anti-APC antibody development and adverse reactions was observed. There was no evidence that the anti-APC antibodies detected represented a specific immune response to DAA therapy. PMID- 20015320 TI - Heparin-induced multiple electrode aggregometry: a potential tool for improvement of heparin-induced thrombocytopenia diagnosis. PMID- 20015321 TI - Enhanced post-clopidogrel platelet reactivity in diabetic patients is independently related to plasma fibrinogen level but not to glycemic control. PMID- 20015323 TI - Molecular characterization of upper urinary tract tumours. AB - OBJECTIVES: To assess gene-expression patterns of BIRC5, FGFR3, IGF2, KRT20, UPK2, EBF1, CDH1, FXYD3, HTERT, TP53, AGR2, HER2 and VEGF, widely known markers of bladder urothelial carcinoma (UC) in upper tract UC, and to determine their value as prognostic factors of tumour progression and cancer-specific survival. PATIENTS AND METHODS: The study included 83 formalin-fixed paraffin-embedded tissue specimens (68 and 15 from patients with UTUC and controls, respectively) collected between 1990 and 2004. Thirteen bladder cancer-related genes were selected from previous reports and analysed by quantitative real-time reverse transcriptase-polymerase chain reaction (qRT-PCR) in all samples. RESULTS: Six genes were over-expressed (BIRC5, FGFR3, KRT20, UPK2, FXYD3 and hTERT) and three under-expressed (AGR2, TP53 and VEGF) in the tumour group (P < 0.05). For four genes (IGF2, EBF1, CDH1 and HER2) there was no statistically significant difference between the tumour and control groups. Overall, 21 patients developed tumour progression and 13 died from UTUC after a mean follow-up of 35.24 months. The 5-year disease-free progression and cancer-specific survival rates were 65.8% and 72.9%, respectively. In a multivariate regression analysis, the independent predictive variable for tumour progression and cancer-specific survival was pathological stage (hazard ratio 3.60, P < 0.001; and 3.73, P < 0.005, respectively), but none of the studied genes were identified as prognostic factors of tumour progression or cancer-specific survival. CONCLUSIONS: Our data suggest that bladder cancer and UTUC share some characteristics, but have differences in gene expression. None of BIRC5, FGFR3, IGF2, KRT20, UPK2, EBF1, CDH1, FXYD3, HTERT, TP53, AGR2, HER2 and VEGF were correlated either tumour progression or survival. PMID- 20015324 TI - Bladder malignancies in children aged <18 years: results from the Surveillance, Epidemiology and End Results database. AB - OBJECTIVE: To present descriptive statistics for bladder tumours in children, calculated from the Surveillance, Epidemiology and End Results (SEER) database, as bladder malignancies are relatively uncommon in children, causing difficulty in understanding their incidence and survival. PATIENTS AND METHODS: The SEER database was interrogated to report the incidence of bladder malignancies in children from birth to 18 years old. Race and sex differences were assessed. The 5-year survival by disease stage at diagnosis and 5-year conditional survival after surviving for 1-3 years is reported for 1973-2003. The incidence and survival rates for bladder embryonal rhabdomyosarcoma were further characterized. RESULTS: We identified 140 cases of bladder cancers in the selected cohort. Papillary urothelial neoplasm of low malignant potential (PUNLMP) and rhabdomyosarcoma comprised 50.7% and 36.4% of the tumours, respectively. The incidence of bladder malignancies significantly increased between 1973 and 2003. Conditional survival calculated for 1 and 2 years after disease diagnosis was 93.6% and 97.5%. Fifty-one cases of embryonal rhabdomyosarcoma were identified. The male to female incidence ratio was approximately 2:1 for these tumours. The 5 year survival rates were 50-80% over the past three decades. CONCLUSIONS: We present the most contemporary survey of the SEER database for the incidence and survival for bladder tumours in children. Depending on the patient's age, PUNLMP and rhabdomyosarcoma predominated in different proportions. The 2-3-year conditional survival and overall survival were excellent. We also identified improving survival for embryonal bladder rhabdomyosarcoma with time. PMID- 20015325 TI - Clinical and inflammatory features of occupational asthma caused by persulphate salts in comparison with asthma associated with occupational rhinitis. AB - BACKGROUND: The relationships between asthma and rhinitis are still a crucial point in respiratory allergy and have scarcely been analysed in occupational setting. We aimed to compare the clinical and inflammatory features of subjects with occupational asthma only (OA) to subjects with OA associated to occupational rhinitis (OAR) caused by persulphate salts. METHODS: The clinical charts of 26 subjects diagnosed in our Unit as respiratory allergy caused by ammonium persulphate (AP), confirmed by specific inhalation challenge (SIC), were reviewed. Twenty-two out of twenty-six patients underwent pre-SIC-induced sputum challenge test (IS) and 24/26 underwent nasal secretion collection and processing. RESULTS: Twelve out of twenty-six patients received a diagnosis of OA only and 14/26 of OAR. Duration of exposure before diagnosis, latency period between the beginning of exposure and asthma symptom onset, basal FEV(1), airway reactivity to methacholine and asthma severity did not differ in the two groups. Eosinophilic inflammation of upper and lower airways characterized both groups. Eosinophil percentage in IS tended to be higher in OAR [11.9 (5.575-13.925)%] than in OA-only [2.95 (0.225-12.5)%] (P = 0.31). Eosinophilia in nasal secretions was present both in subjects with OAR [55 (46-71)%] and in subjects with OA-only [38 (15-73.5)%], without any significant difference. DISCUSSION: Our results indicate that OA because of ammonium persulphate coexists with occupational rhinitis in half of the patients. Unexpectedly, rhinitis did not seem to have an impact on the natural history of asthma. The finding of nasal inflammation in subjects with OA-only without clinical manifestations of rhinitis supports the united airway disease concept in occupational respiratory allergy as a result of persulphates. PMID- 20015326 TI - Relationship of serum cholesterol levels to atopy in the US population. AB - BACKGROUND: Cholesterol promotes Th2 immunity and allergic inflammation in rodents; whether this occurs in humans is unclear. Reports of both direct and inverse associations between serum cholesterol and atopy in different populations suggest that race and/or other demographic variables may modify these relationships. AIMS OF THE STUDY: To determine the relationships between levels of three serum cholesterol measures [total cholesterol (TC), high density lipoprotein-cholesterol (HDL-C), and non-HDL-C] and atopy in a sample representative of the US population. METHODS: Cross-sectional study of 6854 participants aged > or =6 years from the 2005-2006 National Health and Nutrition Examination Survey. RESULTS: In the overall population, adjusted odds ratios (AORs) per two-standard deviation increase in TC and non-HDL-C for biochemical atopy (defined as > or =1 allergen-specific IgE to 19 allergens) were 1.17 [95% confidence interval (CI), 1.00-1.38] and 1.19 (95% CI, 1.03-1.39), respectively. Interactions by race were noted for the two relationships (interaction P = 0.004 and P = 0.009, respectively) with non-Hispanic Whites (NHWs) having direct relationships [TC: AOR 1.27 (95% CI, 1.03-1.57); non-HDL-C: AOR 1.27 (95% CI, 1.03-1.56)] and non-Hispanic Blacks (NHBs) inverse relationships [TC: AOR 0.77 (95% CI, 0.62-0.95); non-HDL-C: AOR 0.86 (95% CI, 0.69-1.08)]. The adjusted HDL-C atopy relationship was nonsignificant for NHWs and inverse for NHBs [AOR 0.77 (95% CI, 0.61-0.96)]. Relationships were independent of body mass index and serum C-reactive protein and unmodified by corticosteroid or statin usage. Results were similar using current hay fever/allergy as the atopy outcome. CONCLUSIONS: There are marked inter-racial differences in the relationship between serum cholesterol and atopy in the US population. PMID- 20015327 TI - Temporal and spatial expression of tumor-associated antigen RCAS1 in pregnant mouse uterus. AB - PROBLEM: The tumor-associated antigen RCAS1 (receptor-binding cancer antigen expressed on SiSo cells) is considered to play a role in the inhibition of maternal immune response during pregnancy, and participates in the initiation of labor and placental detachment. The aim of our study was to investigate the expression of RCAS1 protein in the uteri of normal pregnant mice. METHOD: of study Uteri with fetuses were collected from pregnant ICR mice on days 1.5, 3.5, 5.5, 7.5, and 9.5 p.c., and uterine and placental tissues were obtained separately on days 11.5, 13.5, 15.5, and 17.5 p.c. Samples were examined using real-time (RT)-PCR, Western blotting, and immunohistochemical analyses. RESULTS: In normal pregnant mice, RCAS1 protein mRNA was significantly increased on day 7.5 p.c. Antigen localization was detected in the placenta, decidua, and fetus. CONCLUSION: The results of this study suggest the importance of day 7.5 p.c. for RCAS1 protein expression in connection with placentation as a possible target for future in vivo studies. PMID- 20015328 TI - Circulating and decidual Th17 cell levels in healthy pregnancy. AB - PROBLEM: The Th1/Th2 paradigm has recently been reconstituted to include a third population, Th17 cells. It has been reported that Th2 type immunity is predominantly present in normal pregnancy. However, the level of Th17 cells during pregnancy is still unclear. We investigated the level of peripheral Th17 cells in healthy pregnancy subjects. METHOD: of study To evaluate the levels of Th17 cells, we investigated the proportion of peripheral blood mononuclear cells that produced IL-17 in the first, second, and third trimester pregnancy subjects using flow cytometry. We further studied the proportion of decidual lymphocytes that produced IL-17 in early pregnant subjects. RESULTS: Most of the IL-17 producing cells were CD4(+) T cells. The number of circulating Th17 cells did not change during pregnancy. In a paired t-test of early normal pregnant subjects, the proportion of IL-17(+) decidual lymphocytes was significantly higher than that of peripheral blood lymphocytes. CONCLUSION: Th17 levels in peripheral blood lymphocytes do not change during normal pregnancy. PMID- 20015329 TI - Association study of vascular endothelial growth factor and polymorphisms of its gene with ectopic pregnancy. AB - PROBLEM: In ectopic pregnancy, increased levels of vascular endothelial growth factor are present. The aims of this study were to determine the association between -634C/G, -460T/C, and +936C/T vascular endothelial growth factor (VEGF) polymorphisms and ectopic pregnancy, and to determine whether serum levels of VEGF were affected by genetic factors. METHOD: of study This is a case-control study wherein 74 women with a history of ectopic pregnancy in a tertiary care center were compared to 134 post-menopausal controls with two pregnancies and no ectopic pregnancy for the genotyping of VEGF polymorphisms. For 35 patients with the diagnosis of ectopic pregnancy, serum concentrations of VEGF were obtained before the treatment. Genotyping of VEGF (-634C/G, -460T/C, and +936C/T) polymorphisms was performed by PCR, followed by endonuclease digestion. ELISA was performed to evaluate the VEGF serum levels. RESULTS: The -634C/G, -460T/C, and +936C/T VEGF polymorphisms were not associated with ectopic pregnancy (P = 0.170, P = 0.285, and P = 0.700, respectively). The serum levels of VEGF were not associated with the genotype of -634C/G, -460T/C, and +936C/T VEGF polymorphisms (P = 0.702; P = 0.347, and P = 0.256, respectively). CONCLUSION: There was no association between ectopic pregnancy and -634C/G, -460T/C, and +936C/T VEGF polymorphisms. There was no correlation between VEGF genotype and the expression of VEGF in blood samples. PMID- 20015330 TI - Female genital tract secretions inhibit herpes simplex virus infection: correlation with soluble mucosal immune mediators and impact of hormonal contraception. AB - PROBLEM: Female genital tract secretions inhibit herpes simplex virus (HSV) infection, however, the intra- and inter-subject variability, contribution of specific mediators, and impact of reproductive hormones have not been defined. METHOD: of study Cervicovaginal lavage (CVL) (n = 89) obtained from nine cyclers and seven women on hormonal contraception (HC), who completed between three and eight weekly visits, were examined for anti-herpes simplex virus activity and concentrations of mediators. RESULTS: The CVL inhibited HSV infection by a mean value of approximately 57% during the follicular or luteal phase, but only by 36% in hormonal contraceptive users. Human neutrophil peptides 1-3 (HNP1-3) (P = 0.03), IL-8 (P = 0.003), lactoferrin (P = 0.005), lysozyme (P = 0.003), IgA (P = 0.002), and IgG (P = 0.02) correlated with antiviral activity. Intra-subject and inter-subject variability was observed, suggesting that factors other than hormones contribute to innate defense. CONCLUSION: Endogenous antimicrobial activity may provide a biomarker of healthy mucosal immunity and may be reduced in the setting of HC. However, larger prospective studies are needed. PMID- 20015331 TI - Evidence of mitochondrial dysfunction in obese adolescents. AB - AIM: Although obesity and weight gain generally are anticipated to be caused by an imbalance between energy intake and energy expenditure, the significance of thyroid hormones (TH) remains unclear. Examination of mitochondrial function may reflect intracellular thyroid hormone effect and elucidate whether a lower metabolic rate is present. METHODS: In a group of 34 obese adolescents (age <16 years and body mass index above the age-related 95th percentile), and an age- and gender-matched group of 32 lean adolescent, thyroid stimulating hormone (TSH) and basal oxygen consumption were measured and mitochondrial function in peripheral blood monocytes was determined by flow cytometry. RESULTS: Significant increase in TSH (3.06 +/- 1.56 mU/L vs. 2.33 +/- 0.91 mU/L, p < 0.05) and a decrease in VO2 (129 +/- 16 mL O2/m(2)*min vs. 146 +/- 15 mL O2/m(2)*min, p < 0.05) were observed in obese adolescents compared with lean adolescents. Flow cytometry analysis demonstrated a lower mitochondrial mass (6385 +/- 1962 a.u. vs. 7608 +/- 2328 a.u., p < 0.05) and mitochondrial membrane potential (11426 +/- 3861 a.u. vs. 14017 +/- 5536 a.u., p < 0.05) in obese adolescents compared with lean adolescents. These results are even more pronounced in adolescents with obese mothers. CONCLUSION: In obese adolescents, the increased TSH and lowered VO2 propose a lowered basal metabolic rate and the impaired mitochondrial function suggests a decreased thyroid hormone stimulation of mitochondrial energy production. The maternal in-heritage is suggestive of a basal metabolic defect or mitochondrial resistance for TH. PMID- 20015332 TI - Development of fibroblast cell lines from the cow used to sequence the bovine genome. PMID- 20015333 TI - Characterization of the porcine transferrin gene (TF) and its association with disease severity following an experimental Actinobacillus pleuropneumoniae infection. AB - Transferrin (TF)-mediated provision of iron is essential for a productive infection by many bacterial pathogens, and iron-depletion of TF is a first line defence against bacterial infections. Therefore, the transferrin (TF) gene can be considered a candidate gene for disease resistance. We obtained the complete DNA sequence of the porcine TF gene, which spans 40 kb and contains 17 exons. We identified polymorphisms on a panel of 10 different pig breeds. Comparative intra and interbreed sequence analysis revealed 62 polymorphisms in the TF gene including one microsatellite. Ten polymorphisms were located in the coding sequence of the TF gene. Four SNPs (c.902A>T, c.980G>A, c.1417A>G, c.1810A>C) were predicted to cause amino acid exchanges (p.Lys301Ile, p.Arg327Lys, p.Lys473Glu, p.Asn604His). We performed association analyses using six selected TF markers and 116 pigs experimentally infected with Actinobacillus pleuropneumoniae serotype 7. The analysis showed breed-specific TF allele frequencies. In German Landrace, we found evidence for a possible association of the severity of A. pleuropneumoniae infection with TF genotypes. PMID- 20015334 TI - Pioneering bacterial and algal communities and potential extracellular enzyme activities of stream biofilms. AB - Microbial biofilms are important for the turnover of organic matter in small streams. A rapid colonization of the epilithic surface will become more important given the predicted increase of flood events. Here, we elucidated the pioneering community structure and activity of stream biofilms. Colonization of glass slides exposed in a small stream for 1, 4, 8, 12, and 24 h was compared with those exposed for 7 days or 5 months. Forty thousand microbial cells and 10 algae cm( 2) attached to the glass slides within 1 h of exposure. Catalyzed reporter deposition-FISH demonstrated that the pioneer community that settled within 12 h was dominated by Cytophago-Flavobacteria. Later stages were characterized by an enrichment of Gammaproteobacteria and Betaproteobacteria. However, a major fraction of the detected bacterial cells could not be identified beyond the domain level. Green algae dominated the pioneering algal groups, but were outnumbered by filamentous algae after the attachment period. Potential activity of alkaline phosphatase was already detected after 4 h, beta-glucosidase after 8 h, and beta-xylosidase only after 7 days of biofilm formation. Thus, biofilm formation occurred rapidly and the functionality of the assemblages was evident within a few hours. However, the ratios of beta-xylosidase : beta-glucosidase suggested that the initial biofilms relied more on autochthonous than on allochthonous carbon sources in contrast to mature biofilms. PMID- 20015335 TI - Molecular diversity of arbuscular mycorrhizal fungi and their distribution patterns related to host-plants and habitats in a hot and arid ecosystem, southwest China. AB - The communities of arbuscular mycorrhizal fungi (AMF) colonizing the roots of Bothriochloa pertusa, Cajanus cajan and Heteropogon contortus in a fallow land (FL) and an undisturbed land (UL) were characterized. The large subunit rDNA genes of AMF from roots were amplified and cloned. A total of 2353 clones were screened by restriction fragment length polymorphism, and 428 clones were subsequently sequenced. A total of 393 AMF sequences, which were grouped into 100 operational taxonomic units, were obtained. Phylogenetic analysis revealed that the AMF sequences belonged to Glomus, Acaulospora and Scutellospora, and that Glomus was the dominant genus. Of the 393 AMF sequences, 81% were novel. The diversity of AMF colonizing the same plant species was higher in the UL than in the FL, which confirmed strongly from the molecular evidence that soil disturbance reduced AMF population and species richness. The results revealed that AMF communities were significantly different among host-plant species and between the two habitats. The similarity of AMF communities colonizing different plant species within a habitat was higher than that of the same plant species from different habitats. The molecular evidence supported our previous hypothesis based on morphological analyses that AMF communities were more influenced by habitats compared with host preference. PMID- 20015336 TI - A weak DD-carboxypeptidase activity explains the inability of PBP 6 to substitute for PBP 5 in maintaining normal cell shape in Escherichia coli. AB - Penicillin-binding protein (PBP) 5 plays a critical role in maintaining normal cellular morphology in mutants of Escherichia coli lacking multiple PBPs. The most closely related homologue, PBP 6, is 65% identical to PBP 5, but is unable to substitute for PBP 5 in returning these mutants to their wild-type shape. The relevant differences between PBPs 5 and 6 are localized in a 20-amino acid stretch of domain I in these proteins, which includes the canonical KTG motif at the active site. We determined how these differences affected the enzymatic properties of PBPs 5 and 6 toward beta-lactam binding and the binding and hydrolysis of two peptide substrates. We also investigated the enzymatic properties of recombinant fusion proteins in which active site segments were swapped between PBPs 5 and 6. The results suggest that the in vivo physiological role of PBP 5 is distinguished from PBP 6 by the higher degree of DD carboxypeptidase activity of the former. PMID- 20015337 TI - Increased synthesis of signaling molecules coincides with reversible inhibition of nucleolytic activity during postirradiation recovery of Deinococcus radiodurans. AB - Deinococcus radiodurans tolerates extensive DNA damage and exhibits differential expression of various genes associated with the growth of the organism and DNA repair. In cells treated with gamma radiation, the levels of cyclic AMP (cAMP) and ATP increased rapidly by differentially regulating adenylyl cyclase (AC) and 2'3' cAMP phosphodiesterase. The levels of cAMP, ATP, AC and protein kinases were high when phosphodiesterase activity was low. These cells exhibited in vivo inhibition of nucleolytic function by reversible protein phosphorylation and contained the comparatively higher levels of total phosphoproteins. We suggest that Deinococcus, a prokaryote, uses DNA damage-induced signaling mechanism as evidenced by gamma radiation-induced synthesis of secondary messengers and signaling enzymes. PMID- 20015338 TI - Identification of a gene coding for a deglycosylating enzyme in Hypocrea jecorina. AB - An enzyme with mannosyl glycoprotein endo-N-acetyl-beta-D-glucosaminidase (ENGase)-type activity was partially purified from the extracellular medium of the mould Hypocrea jecorina (Trichoderma reesei). Internal peptides were generated and used to identify the gene in the T. reesei genome. The active enzyme is processed both at the N- and at the C-terminus. High-mannose-type glycoproteins are good substrates, whereas complex-type glycans are not hydrolysed. The enzyme represents the first fungal member of glycoside hydrolase family 18 with ENGase-type activity. Bacterial ENGases and the fungal chitinases belonging to the same family show very low homology with Endo T. Database searches identify several highly homologous genes in fungi and the activity is also found within other Trichoderma species. This ENGase activity, not coregulated with cellulase production, could be responsible for the extensive N deglycosylation observed for several T. reesei cellulases. PMID- 20015339 TI - Diffuse xanthomatosis as a presenting feature of multiple myeloma. PMID- 20015340 TI - Structural insight into the membrane insertion of tail-anchored proteins by Get3. AB - Tail anchored (TA) proteins, which are important for numerous cellular processes, are defined by a single transmembrane domain (TMD) near the C-terminus. The membrane insertion of TA proteins is mediated by the highly conserved ATPase Get3. Here we report the crystal structures of Get3 in ADP-bound and nucleotide free forms at 3.0 A and 2.8 A resolutions, respectively. Get3 consists of a nucleotide binding domain and a helical domain. Both structures exhibit a Zn(2+) mediated homodimer in a head-to-head orientation, representing an open dimer conformation. Our cross-link experiments indicated the closed dimer-stimulating ATP hydrolysis, which might be coupled with TA-protein release. Further, our coexpression-based binding assays using a model TA protein Sec22p revealed the direct interaction between the helical domain of Get3 and the Sec22p TMD. This interaction is independent of ATP and dimer formation. Finally, we propose a structural mechanism that links ATP hydrolysis with the TA-protein insertion mediated by the conserved DTAPTGH motif. PMID- 20015342 TI - Foreword to special section: groundwater in Africa. PMID- 20015341 TI - Genetic disruptions of Drosophila Pavlovian learning leave extinction learning intact. AB - Individuals who experience traumatic events may develop persistent posttraumatic stress disorder. Patients with this disorder are commonly treated with exposure therapy, which has had limited long-term success. In experimental neurobiology, fear extinction is a model for exposure therapy. In this behavioral paradigm, animals are repeatedly exposed in a safe environment to the fearful stimulus, which leads to greatly reduced fear. Studying animal models of extinction already has lead to better therapeutic strategies and development of new candidate drugs. Lack of a powerful genetic model of extinction, however, has limited progress in identifying underlying molecular and genetic factors. In this study, we established a robust behavioral paradigm to study the short-term effect (acquisition) of extinction in Drosophila melanogaster. We focused on the extinction of olfactory aversive 1-day memory with a task that has been the main workhorse for genetics of memory in flies. Using this paradigm, we show that extinction can inhibit each of two genetically distinct forms of consolidated memory. We then used a series of single-gene mutants with known impact on associative learning to examine the effects on extinction. We find that extinction is intact in each of these mutants, suggesting that extinction learning relies on different molecular mechanisms than does Pavlovian learning. PMID- 20015343 TI - Modeling basin- and plume-scale processes of CO2 storage for full-scale deployment. AB - Integrated modeling of basin- and plume-scale processes induced by full-scale deployment of CO(2) storage was applied to the Mt. Simon Aquifer in the Illinois Basin. A three-dimensional mesh was generated with local refinement around 20 injection sites, with approximately 30 km spacing. A total annual injection rate of 100 Mt CO(2) over 50 years was used. The CO(2)-brine flow at the plume scale and the single-phase flow at the basin scale were simulated. Simulation results show the overall shape of a CO(2) plume consisting of a typical gravity-override subplume in the bottom injection zone of high injectivity and a pyramid-shaped subplume in the overlying multilayered Mt. Simon, indicating the important role of a secondary seal with relatively low-permeability and high-entry capillary pressure. The secondary-seal effect is manifested by retarded upward CO(2) migration as a result of multiple secondary seals, coupled with lateral preferential CO(2) viscous fingering through high-permeability layers. The plume width varies from 9.0 to 13.5 km at 200 years, indicating the slow CO(2) migration and no plume interference between storage sites. On the basin scale, pressure perturbations propagate quickly away from injection centers, interfere after less than 1 year, and eventually reach basin margins. The simulated pressure buildup of 35 bar in the injection area is not expected to affect caprock geomechanical integrity. Moderate pressure buildup is observed in Mt. Simon in northern Illinois. However, its impact on groundwater resources is less than the hydraulic drawdown induced by long-term extensive pumping from overlying freshwater aquifers. PMID- 20015344 TI - Understanding the impacts of anisotropy on the extent of drawdown. PMID- 20015345 TI - HIV prevention is not enough: child survival in the context of prevention of mother to child HIV transmission. AB - Clinical and epidemiologic research has identified increasingly effective interventions to reduce mother to child HIV transmission in resource-limited settings These scientific breakthroughs have been implemented in some programmes, although much remains to be done to improve coverage and quality of these programmes. But prevention of HIV transmission is not enough. It is necessary also to consider ways to improve maternal health and protect child survival.A win win approach is to ensure that all pregnant and lactating women with CD4 counts of <350 cells/mm3 have access to antiretroviral therapy. On its own, this approach will substantially improve maternal health and markedly reduce mother to child HIV transmission during pregnancy and delivery and through breastfeeding. This approach can be combined with additional interventions for women with higher CD4 counts, either extended prophylaxis to infants or extended regimens of antiretroviral drugs to women, to reduce transmission even further.Attempts to encourage women to abstain from all breastfeeding or to shorten the optimal duration of breastfeeding have led to increases in mortality among both uninfected and infected children. A better approach is to support breastfeeding while strengthening programmes to provide antiretroviral therapy for pregnant and lactating women who need it and offering antiretroviral drug interventions through the duration of breastfeeding. This will lead to reduced HIV transmission and will protect the health of women without compromising the health and well being of infants and young children. PMID- 20015346 TI - The adequacy of policy responses to the treatment needs of South Africans living with HIV (1999-2008): a case study. AB - INTRODUCTION: South Africa has the largest HIV/AIDS epidemic of any country in the world. CASE DESCRIPTION: National antiretroviral therapy (ART) policy is examined over the period of 1999 to 2008, which coincided with the government of President Thabo Mbeki and his Minister of Health, Dr Manto Tshabalala-Msimang. The movement towards a national ART programme in South Africa was an ambitious undertaking, the likes of which had not been contemplated before in public health in Africa. DISCUSSION AND EVALUATION: One million AIDS-ill individuals were targeted to be enrolled in the ART programme by 2007/08. Fewer than 50% of eligible individuals were enrolled. This failure resulted from lack of political commitment and inadequate public health system capacity. The human and economic costs of this failure are large and sobering. CONCLUSIONS: The total lost benefits of ART not reaching the people who need it are estimated at 3.8 million life years for the period, 2000 to 2005. The economic cost of those lost life years over this period has been estimated at more than US$15 billion. PMID- 20015347 TI - Psychoeducation for depression, anxiety and psychological distress: a meta analysis. AB - BACKGROUND: Given the high prevalence and burden associated with depression and anxiety disorders and the existence of treatment barriers, there is a clear need for brief, inexpensive and effective interventions such as passive psychoeducational interventions. There are no published meta-analyses of the effectiveness of passive psychoeducation in reducing symptoms of depression, anxiety or psychological distress. METHODS: Cochrane, PsycInfo and PubMed databases were searched in September 2008. Additional materials were obtained from reference lists. Papers describing passive psychoeducational interventions for depression, anxiety and psychological distress were included if the research design was a randomized controlled trial and incorporated an attention placebo, no intervention or waitlist comparison group. RESULTS: In total, 9010 abstracts were identified. Of these, five papers which described four research studies targeting passive psychoeducation for depression and psychological distress met the inclusion criteria. The pooled standardized-effect size (four studies, four comparisons) for reduced symptoms of depression and psychological distress at post-intervention was d = 0.20 (95% confidence interval: 0.01-0.40; Z = 2.04; P = 0.04; the number needed to treat: 9). Heterogeneity was not significant among the studies (I2 = 32.77, Q:4.46; P = 0.22). CONCLUSIONS: Although it is commonly believed that psychoeducation interventions are ineffective, this meta-analysis revealed that brief passive psychoeducational interventions for depression and psychological distress can reduce symptoms. Brief passive psychoeducation interventions are easy to implement, can be applied immediately and are not expensive. They may offer a first-step intervention for those experiencing psychological distress or depression and might serve as an initial intervention in primary care or community models. The findings suggest that the quality of psychoeducation may be important. PMID- 20015348 TI - A novel Bifidobacterium infantis-mediated TK/GCV suicide gene therapy system exhibits antitumor activity in a rat model of bladder cancer. AB - Bladder cancer is the ninth most common malignancy in the world. Successful clinical management remains a challenge. In order To search for novel targeted and efficacious treatment, we sought to investigate anti-tumor activity of BI-TK suicide gene therapy system in a rat model of bladder tumors. We first constructed and tested an anaerobic Bifidobacterium infantis-mediated thymidine kinase (BI-TK) suicide gene therapy system. To test the in vivo efficacy of this system, we established a rat model of bladder tumors, which was induced by N methyl-nitrosourea perfusion. Bifidobacterium infantis containing the HSV-TK (i.e., BI-TK) were constructed by transformation of recombinant plasmid pGEX - TK. The engineered BI-TK was injected into tumor-bearing rats via tail vein, followed by intraperitoneal injection of ganciclovir (GCV). Using the rat model of bladder tumors, we found that bladder tumor burdens were significantly lower in the rats treated with BI-TK/GCV group than that treated with normal saline control group (p <0.05). While various degrees of apoptosis of the tumor cells were detected in all groups using in situ TUNEL assay, apoptosis was mostly notable in the BI-TK/GCV treatment group. Immunohistochemical staining further demonstrated that the BI-TK/GCV treatment group had the highest level of caspase3 protein expression than that of the empty plasmid group and normal saline group (p < 0.05). Thus, our results demonstrate that the Bifidobacterium infantis mediated TK/GCV suicide gene therapy system can effectively inhibit rat bladder tumor growth, possibly through increasing caspase 3 expression and inducing apoptosis. PMID- 20015349 TI - Genome-wide nucleosome mapping of Plasmodium falciparum reveals histone-rich coding and histone-poor intergenic regions and chromatin remodeling of core and subtelomeric genes. AB - BACKGROUND: Epigenetic modifications of histones and regulation of chromatin structure have been implicated in regulation of virulence gene families in P. falciparum. To better understand chromatin-mediated gene regulation, we used a high-density oligonucleotide microarray to map the position and enrichment of nucleosomes across the entire genome of P. falciparum at three time points of the intra-erythrocytic developmental cycle (IDC) in vitro. We used an unmodified histone H4 antibody for chromatin immunoprecipitation of nucleosome-bound DNA. RESULTS: We observed generally low nucleosomal occupancy of intergenic regions and higher occupancy of protein coding regions. In contract to the overall small fluctuation of nucleosomal occupancy in most coding regions throughout the IDC, subtelomeric genes encoding surface proteins such as var and rif, as well as some core chromosomal genes such as transcription factors, showed large changes in chromatin structure. Telomeres harbored a region with the highest nucleosomal occupancy of the genome and also exhibited large changes with higher nucleosomal occupancy at schizont stages. While many of these subtelomeric genes were previously shown to be modified by H3K9 trimethylation, we also identified some housekeeping genes in core chromosome regions that showed extensive changes in chromatin structure but do not contain this modification. tRNA and basal transcription factor genes showed low nucleosomal occupancy at all times, suggesting of an open chromatin structure that might be permissive for constitutively high levels of expression. Generally, nucleosomal occupancy was not correlated with the steady-state mRNA levels. Several var genes were exceptions: the var gene with the highest expression level showed the lowest nucleosomal occupancy, and selection of parasites for var2CSA expression resulted in lower nucleosomal occupancy at the var2CSA locus. We identified nucleosome free regions in intergenic regions that may serve as transcription start sites or transcription factor binding sites. Using the nucleosomal occupancy data as the baseline, we further mapped the genome-wide enrichment of H3K9 acetylation and detected general enrichment of this mark in intergenic regions. CONCLUSIONS: These data on nucleosome enrichment changes add to our understanding of the influence of chromatin structure on the regulation of gene expression. Histones are generally enriched in coding regions, and relatively poor in intergenic regions. Histone enrichment patterns allow for identification of new putative gene-coding regions. Most genes do not show correlation between chromatin structure and steady-state mRNA levels, indicating the dominant roles of other regulatory mechanisms. We present a genome-wide nucleosomal occupancy map, which can be used as a reference for future experiments of histone modification mapping. PMID- 20015350 TI - The sonic hedgehog signaling pathway is reactivated in human renal cell carcinoma and plays orchestral role in tumor growth. AB - BACKGROUND: Human clear cell renal cell carcinoma (CRCC) remains resistant to therapies. Recent advances in Hypoxia Inducible Factors (HIF) molecular network led to targeted therapies, but unfortunately with only limited clinical significance. Elucidating the molecular processes involved in kidney tumorigenesis and resistance is central to the development of improved therapies, not only for kidney cancer but for many, if not all, cancer types. The oncogenic PI3K/Akt, NF-kB and MAPK pathways are critical for tumorigenesis. The sonic hedgehog (SHH) signaling pathway is crucial to normal development. RESULTS: By quantitative RT-PCR and immunoblot, we report that the SHH signaling pathway is constitutively reactivated in tumors independently of the von Hippel-Lindau (VHL) tumor suppressor gene expression which is inactivated in the majority of CRCC. The inhibition of the SHH signaling pathway by the specific inhibitor cyclopamine abolished CRCC cell growth as assessed by cell counting, BrdU incorporation studies, fluorescence-activated cell sorting and beta-galactosidase staining. Importantly, inhibition of the SHH pathway induced tumor regression in nude mice through inhibition of cell proliferation and neo-vascularization, and induction of apoptosis but not senescence assessed by in vivo studies, immunoblot and immunohistochemistry. Gli1, cyclin D1, Pax2, Lim1, VEGF, and TGF-beta were exclusively expressed in tumors and were shown to be regulated by SHH, as evidenced by immunoblot after SHH inhibition. Using specific inhibitors and immunoblot, the activation of the oncogenic PI3K/Akt, NF-kB and MAPK pathways was decreased by SHH inhibition. CONCLUSIONS: These findings support targeting SHH for the treatment of CRCC and pave the way for innovative and additional investigations in a broad range of cancers. PMID- 20015351 TI - A dose-controlled system for air-liquid interface cell exposure and application to zinc oxide nanoparticles. AB - BACKGROUND: Engineered nanoparticles are becoming increasingly ubiquitous and their toxicological effects on human health, as well as on the ecosystem, have become a concern. Since initial contact with nanoparticles occurs at the epithelium in the lungs (or skin, or eyes), in vitro cell studies with nanoparticles require dose-controlled systems for delivery of nanoparticles to epithelial cells cultured at the air-liquid interface. RESULTS: A novel air liquid interface cell exposure system (ALICE) for nanoparticles in liquids is presented and validated. The ALICE generates a dense cloud of droplets with a vibrating membrane nebulizer and utilizes combined cloud settling and single particle sedimentation for fast (~10 min; entire exposure), repeatable (<12%), low-stress and efficient delivery of nanoparticles, or dissolved substances, to cells cultured at the air-liquid interface. Validation with various types of nanoparticles (Au, ZnO and carbon black nanoparticles) and solutes (such as NaCl) showed that the ALICE provided spatially uniform deposition (<1.6% variability) and had no adverse effect on the viability of a widely used alveolar human epithelial-like cell line (A549). The cell deposited dose can be controlled with a quartz crystal microbalance (QCM) over a dynamic range of at least 0.02-200 mug/cm(2). The cell-specific deposition efficiency is currently limited to 0.072 (7.2% for two commercially available 6-er transwell plates), but a deposition efficiency of up to 0.57 (57%) is possible for better cell coverage of the exposure chamber. Dose-response measurements with ZnO nanoparticles (0.3-8.5 mug/cm(2)) showed significant differences in mRNA expression of pro-inflammatory (IL-8) and oxidative stress (HO-1) markers when comparing submerged and air liquid interface exposures. Both exposure methods showed no cellular response below 1 mug/cm(2 )ZnO, which indicates that ZnO nanoparticles are not toxic at occupationally allowed exposure levels. CONCLUSION: The ALICE is a useful tool for dose-controlled nanoparticle (or solute) exposure of cells at the air-liquid interface. Significant differences between cellular response after ZnO nanoparticle exposure under submerged and air-liquid interface conditions suggest that pharmaceutical and toxicological studies with inhaled (nano-)particles should be performed under the more realistic air-liquid interface, rather than submerged cell conditions. PMID- 20015352 TI - Global transcriptional response after exposure of fission yeast cells to ultraviolet light. AB - BACKGROUND: In many cell types, including the fission yeast Schizosaccharomyces pombe, a set of checkpoints are induced by perturbations of the cell cycle or by DNA damage. Many of the checkpoint responses include a substantial change of the transcriptional pattern. As part of characterising a novel G1/S checkpoint in fission yeast we have investigated whether a transcriptional response is induced after irradiation with ultraviolet light. RESULTS: Microarray analyses were used to measure the global transcription levels of all open reading frames of fission yeast after 254 nm ultraviolet irradiation, which is known to induce a G1/S checkpoint. We discovered a surprisingly weak transcriptional response, which is quite unlike the marked changes detected after some other types of treatment and in several other checkpoints. Interestingly, the alterations in gene expression after ultraviolet irradiation were not similar to those observed after ionising radiation or oxidative stress. Pathway analysis suggests that there is little systematic transcriptional response to the irradiation by ultraviolet light, but a marked, coordinated transcriptional response was noted on progression of the cells from G1 to S phase. CONCLUSION: There is little response in fission yeast to ultraviolet light at the transcriptional level. Amongst the genes induced or repressed after ultraviolet irradiation we found none that are likely to be involved in the G1/S checkpoint mechanism, suggesting that the checkpoint is not dependent upon transcriptional regulation. PMID- 20015353 TI - Risk factors for childhood obesity at age 5: analysis of the millennium cohort study. AB - BACKGROUND: Weight at age 5 is a predictor for future health of the individual. This study examines risk factors for childhood obesity with a focus on ethnicity. METHODS: Data from the Millennium Cohort study were used. 17,561 singleton children of White/European (n = 15,062), Asian (n = 1,845) or African (n = 654) background were selected. Logistic regression and likelihood ratio tests were used to examine factors associated with obesity at age 5. All participants were interviewed in their own homes. The main exposures examined included; Birth weight, sedentary lifestyle, family health behaviours, ethnicity, education and income. RESULTS: Children with a sedentary lifestyle, large at birth, with high risk family health behaviours (overweight mothers, smoking near the child, missing breakfast) and from a family with low income or low educational attainment, were more likely to be obese regardless of ethnicity. Feeding solid food before 3 months was associated with obesity in higher income White/European families. Even when controlling for socioeconomic status, ethnic background is an important independent risk factor for childhood obesity [Odds ratio of obesity; was 1.7 (95%CI: 1.2-2.3) for Asian and 2.7 (95%CI: 1.9-3.9) for African children, compared to White/European]. The final adjusted model suggests that increasing income does not have a great impact on lowering obesity levels, but that higher academic qualifications are associated with lower obesity levels [Odds of obesity: 0.63 (95%CI: 0.52-0.77) if primary carer leaves school after age 16 compared at age 16]. CONCLUSIONS: Education of the primary carer is an important modifiable factor which can be targeted to address rising obesity levels in children. Interventions should be family centred supporting and showing people how they can implement lifestyle changes in their family. PMID- 20015354 TI - Better assessment of physical function: item improvement is neglected but essential. AB - INTRODUCTION: Physical function is a key component of patient-reported outcome (PRO) assessment in rheumatology. Modern psychometric methods, such as Item Response Theory (IRT) and Computerized Adaptive Testing, can materially improve measurement precision at the item level. We present the qualitative and quantitative item-evaluation process for developing the Patient Reported Outcomes Measurement Information System (PROMIS) Physical Function item bank. METHODS: The process was stepwise: we searched extensively to identify extant Physical Function items and then classified and selectively reduced the item pool. We evaluated retained items for content, clarity, relevance and comprehension, reading level, and translation ease by experts and patient surveys, focus groups, and cognitive interviews. We then assessed items by using classic test theory and IRT, used confirmatory factor analyses to estimate item parameters, and graded response modeling for parameter estimation. We retained the 20 Legacy (original) Health Assessment Questionnaire Disability Index (HAQ-DI) and the 10 SF-36's PF 10 items for comparison. Subjects were from rheumatoid arthritis, osteoarthritis, and healthy aging cohorts (n = 1,100) and a national Internet sample of 21,133 subjects. RESULTS: We identified 1,860 items. After qualitative and quantitative evaluation, 124 newly developed PROMIS items composed the PROMIS item bank, which included revised Legacy items with good fit that met IRT model assumptions. Results showed that the clearest and best-understood items were simple, in the present tense, and straightforward. Basic tasks (like dressing) were more relevant and important versus complex ones (like dancing). Revised HAQ-DI and PF 10 items with five response options had higher item-information content than did comparable original Legacy items with fewer response options. IRT analyses showed that the Physical Function domain satisfied general criteria for unidimensionality with one-, two-, three-, and four-factor models having comparable model fits. Correlations between factors in the test data sets were > 0.90. CONCLUSIONS: Item improvement must underlie attempts to improve outcome assessment. The clear, personally important and relevant, ability-framed items in the PROMIS Physical Function item bank perform well in PRO assessment. They will benefit from further study and application in a wider variety of rheumatic diseases in diverse clinical groups, including those at the extremes of physical functioning, and in different administration modes. PMID- 20015355 TI - Identification of horse chestnut coat color genotype using SNaPshot. AB - BACKGROUND: The Cantabrian Coast horse breeds of the Iberian Peninsula have mainly black or bay colored coats, but alleles responsible for a chestnut coat color run in these breeds and occasionally, chestnut horses are born. Chestnut coat color is caused by two recessive alleles, e and e(a), of the melanocortin-1 receptor gene, whereas the presence of the dominant, wild-type E allele produces black or bay coat horses. Because black or bay colored coats are considered as the purebred phenotype for most of the breeds from this region, it is important to have a fast and reliable method to detect alleles causing chestnut coat color in horses. FINDINGS: In order to assess coat color genotype in reproductive animals with a view to avoiding those bearing chestnut alleles, we have developed a reliable, fast and cost-effective screening device which involves Single Nucleotide Polymorphism (SNP) detection based on SNaPshot((R) )(Applied Biosystems) methodology. We have applied this method to four native breeds from the Iberian Cantabrian Coast: Pottoka and Jaca Navarra pony breeds, in which only black or bay coats are acceptable, and Euskal Herriko Mendiko Zaldia and Burguete heavy breeds, in which chestnut coats are acceptable. The frequency of the chestnut alleles ranged between f = 0.156-0.322 in pony breeds and between f = 0.604-0.716 in heavy breeds. CONCLUSIONS: This study demonstrates the usefulness of the DNA methodology reported herein as a device for identifying chestnut alleles; the methodology constitutes a valuable tool for breeders to decrease the incidence of chestnut animals among Cantabrian Coast pony breeds. PMID- 20015356 TI - The relationship between chronic type III acromioclavicular joint dislocation and cervical spine pain. AB - BACKGROUND: This study was aimed at evaluating whether or not patients with chronic type III acromioclavicular dislocation develop cervical spine pain and degenerative changes more frequently than normal subjects. METHODS: The cervical spine of 34 patients with chronic type III AC dislocation was radiographically evaluated. Osteophytosis presence was registered and the narrowing of the intervertebral disc and cervical lordosis were evaluated. Subjective cervical symptoms were investigated using the Northwick Park Neck Pain Questionnaire (NPQ). One-hundred healthy volunteers were recruited as a control group. RESULTS: The rate and distribution of osteophytosis and narrowed intervertebral disc were similar in both of the groups. Patients with chronic AC dislocation had a lower value of cervical lordosis. NPQ score was 17.3% in patients with AC separation (100% = the worst result) and 2.2% in the control group (p < 0.05). An inverse significant nonparametric correlation was found between the NPQ value and the lordosis degree in the AC dislocation group (p = 0.001) wheras results were not correlated (p = 0.27) in the control group. CONCLUSIONS: Our study shows that chronic type III AC dislocation does not interfere with osteophytes formation or intervertebral disc narrowing, but that it may predispose cervical hypolordosis. The higher average NPQ values were observed in patients with chronic AC dislocation, especially in those that developed cervical hypolordosis. PMID- 20015357 TI - The incidence of sub-optimal sedation in the ICU: a systematic review. AB - INTRODUCTION: Patients in intensive care units (ICUs) are generally sedated for prolonged periods. Over-sedation and under-sedation both have negative effects on patient safety and resource use. We conducted a systematic review of the literature in order to establish the incidence of sub-optimal sedation (both over and under-sedation) in ICUs. METHODS: We searched Medline, Embase and CINAHL (Cumulative Index to Nursing and Allied Health Literature) online literature databases from 1988 to 15 May 2008 and hand-searched conferences. English language studies set in the ICU, in sedated adult humans on mechanical ventilation, which reported the incidence of sub-optimal sedation, were included. All abstracts were reviewed twice by two independent reviewers, with all conflicts resolved by a third reviewer, to check that they met the review inclusion criteria. Full papers of all included studies were retrieved and were again reviewed twice against inclusion criteria. Data were doubly extracted. Study aims, design, population, comparisons made, and data on the incidence of sub-optimal, optimal, over-sedation or under-sedation were extracted. RESULTS: There was considerable variation between included studies in the definition of optimal sedation and in the scale or method used to assess sedation. Across all included studies, a substantial incidence of sub-optimal sedation was reported, with a greater tendency toward over-sedation. CONCLUSIONS: Our review suggests that improvements in the consistent definition and measurement of sedation may improve the quality of care of patients within the ICU. PMID- 20015358 TI - Phytalgic, a food supplement, vs placebo in patients with osteoarthritis of the knee or hip: a randomised double-blind placebo-controlled clinical trial. AB - INTRODUCTION: The medicinal treatment of osteoarthritis (OA) is mostly symptomatic to relieve pain and incapacity with analgesics and non-steroidal anti inflammatory drugs (NSAIDs), drugs with well-known risks. Complementary medicines might reduce the symptoms of OA and decrease the need for NSAIDs. This study tested the effects of a food supplement, Phytalgic, on pain and function in patients with osteoarthritis and their use of analgesic and NSAIDs. METHODS: A randomized double-blind parallel-groups clinical trial compared Phytalgic (fish oil, vitamin E, Urtica dioica) to a placebo for three months, in 81 patients with OA of the knee or hip using NSAIDs and/or analgesics regularly. The main outcome measures were use of NSAIDs (in Defined Daily Doses per day - DDD/day) or analgesics (in 500 mg paracetamol-equivalent tablets per week (PET/week) measured each month, and Western Ontario-McMaster University Osteo-Arthritis Index (WOMAC) function scales. RESULTS: After three months of treatment, the mean use of analgesics in the active arm (6.5 PET/week) vs. the placebo arm (16.5) was significantly different (P < 0.001) with a group mean difference of -10.0 (95% CI: -4.9 to -15.1). That of NSAIDs in the active arm (0.4 DDD/day) vs the placebo arm (1.0 DDD/day) was significantly different (P = 0.02) with a group mean difference of - 0.7 DDD/day (95% CI: -0.2 to -1.2). Mean WOMAC scores for pain, stiffness and function in the active arm (respectively 86.5, 41.4 and 301.6) vs the placebo arm (resp. 235.3, 96.3 and 746.5) were significantly different (P < 0.001) with group mean differences respectively of -148.8 (95% CI: -97.7 to 199.9), -54.9 (95% CI: -27.9 to -81.9) and -444.8 (95% CI: -269.1 to -620.4). CONCLUSIONS: The food supplement tested appeared to decrease the need for analgesics and NSAIDs and improve the symptoms of osteoarthritis. TRIAL REGISTRATION: Clinicaltrials.gov NCT00666523. PMID- 20015359 TI - Molecular evolution of sex-biased genes in the Drosophila ananassae subgroup. AB - BACKGROUND: Genes with sex-biased expression often show rapid molecular evolution between species. Previous population genetic and comparative genomic studies of Drosophila melanogaster and D. simulans revealed that male-biased genes have especially high rates of adaptive evolution. To test if this is also the case for other lineages within the melanogaster group, we investigated gene expression in D. ananassae, a species that occurs in structured populations in tropical and subtropical regions. We used custom-made microarrays and published microarray data to characterize the sex-biased expression of 129 D. ananassae genes whose D. melanogaster orthologs had been classified previously as male-biased, female biased, or unbiased in their expression and had been studied extensively at the population-genetic level. For 43 of these genes we surveyed DNA sequence polymorphism in a natural population of D. ananassae and determined divergence to the sister species D. atripex and D. phaeopleura. RESULTS: Sex-biased expression is generally conserved between D. melanogaster and D. ananassae, with the majority of genes exhibiting the same bias in the two species. However, about one third of the genes have either gained or lost sex-biased expression in one of the species and a small proportion of genes (approximately 4%) have changed bias from one sex to the other. The male-biased genes of D. ananassae show evidence of positive selection acting at the protein level. However, the signal of adaptive protein evolution for male-biased genes is not as strong in D. ananassae as it is in D. melanogaster and is limited to genes with conserved male-biased expression in both species. Within D. ananassae, a significant signal of adaptive evolution is also detected for female-biased and unbiased genes. CONCLUSIONS: Our findings extend previous observations of widespread adaptive protein evolution to an independent Drosophila lineage, the D. ananassae subgroup. However, the rate of adaptive evolution is not greater for male-biased genes than for female-biased or unbiased genes, which suggests that there are differences in sex-biased gene evolution between the two lineages. PMID- 20015360 TI - Human synthetic lethal inference as potential anti-cancer target gene detection. AB - BACKGROUND: Two genes are called synthetic lethal (SL) if mutation of either alone is not lethal, but mutation of both leads to death or a significant decrease in organism's fitness. The detection of SL gene pairs constitutes a promising alternative for anti-cancer therapy. As cancer cells exhibit a large number of mutations, the identification of these mutated genes' SL partners may provide specific anti-cancer drug candidates, with minor perturbations to the healthy cells. Since existent SL data is mainly restricted to yeast screenings, the road towards human SL candidates is limited to inference methods. RESULTS: In the present work, we use phylogenetic analysis and database manipulation (BioGRID for interactions, Ensembl and NCBI for homology, Gene Ontology for GO attributes) in order to reconstruct the phylogenetically-inferred SL gene network for human. In addition, available data on cancer mutated genes (COSMIC and Cancer Gene Census databases) as well as on existent approved drugs (DrugBank database) supports our selection of cancer-therapy candidates. CONCLUSIONS: Our work provides a complementary alternative to the current methods for drug discovering and gene target identification in anti-cancer research. Novel SL screening analysis and the use of highly curated databases would contribute to improve the results of this methodology. PMID- 20015361 TI - Identifying the Tuskegee Syphilis Study: implications of results from recall and recognition questions. AB - BACKGROUND: This analysis assessed whether Blacks, Whites and Puerto-Rican (PR) Hispanics differed in their ability to identify the Tuskegee Syphilis Study (TSS) via open-ended questions following lead-in recognition and recall questions. METHODS: The Tuskegee Legacy Project (TLP) Questionnaire was administered via a Random-Digit Dial (RDD) telephone survey to a stratified random sample of Black, White and PR Hispanic adults in three U.S. cities. RESULTS: The TLP Questionnaire was administered to 1,162 adults (356 African-Americans, 313 PR Hispanics, and 493 non-Hispanic Whites) in San Juan, PR, Baltimore, MD and New York City, NY. Recall question data revealed: 1) that 89% or more of Blacks, Whites, and PR Hispanics were not able to name or definitely identify the Tuskegee Syphilis Study by giving study attributes; and, 2) that Blacks were the most likely to provide an open-ended answer that identified the Tuskegee Syphilis Study as compared to Whites and PR Hispanics (11.5% vs 6.3% vs 2.9%, respectively) (p /=19, who underwent emergency surgery for acute complications (5 for an abdominal compartment syndrome and 1 for septic shock) followed by continuous perioperative peritoneal lavage and postoperative CVVDH. CVVDH was started within 12 hours after surgery and maintained for at least 72 hours, until the multiorgan dysfunction syndrome improved. Samples were collected from serum, peritoneal lavage fluid and CVVDH dialysate for cytokine assay. Apache II scores were measured daily and their association with cytokine levels was assessed. RESULTS: All six patients tolerated CVVDH well, and the procedure lasted a mean 6 days (range, 3-12). Five patients survived and one died of Acinetobacter infection after surgery (mortality rate 16.6%). The mean APACHE II score was >/= 19 (range 19-22) before laparotomy and decreased significantly during peritoneal lavage and postoperative CVVDH (P = 0.013 by matched-pairs Students t-test). The decrease in cytokine concentrations in serum and lavage fluid was associated with the decrease in APACHE II scores and high interleukin 6 (IL-6) and tumor necrosis factor (TNF) concentrations in the hemofiltrate. CONCLUSION: In critically ill patients with abdominal compartment syndrome, septic shock or high APACHE II scores related to severe acute pancreatitis, combining emergency laparotomy with continuous perioperative peritoneal lavage followed by postoperative CVVHD effectively reduces the local and systemic cytokines responsible for multiorgan dysfunction syndrome thus improving patients' outcome. PMID- 20015377 TI - Assessment of sperm quality traits in relation to fertility in boar semen. AB - BACKGROUND: Several studies have been published where sperm plasma membrane integrity correlated to fertility. In this study we describe a simple fluorometer based assay where we monitored the fluorescence intensity of artificially membrane-ruptured spermatozoa with a fixed time staining with fluorescent DNA dyes. METHODS: Membrane-impermeant fluorescent dyes Hoechst 33258 (H258) and propidium iodide (PI) were used to measure the fluorescence of the nucleus in artificially membrane ruptured spermatozoa and membrane-permeant dye Hoechst 33342 (H342) was used to measure fluorescence of intact spermatozoa. The concentration of spermatozoa in insemination doses varied from 31.2x10(6)/ml to 50x10(6)/ml and the average value was 35x10(6)/ml. Each boar was represented by three consecutive ejaculates, collected at weekly intervals. Nonreturn rate within 60 days of first insemination (NR%) and litter size (total number of piglets born) of multiparous farrowings were used as fertility measures. RESULTS: Sperm fluorescence intensity of H258 and H342, but not the fluorescence intensity of PI-stained spermatozoa correlated significantly with the litter size of multiparous farrowings, values being r=-0.68 (P<0.01) for H258, r=-0.69 (P<0.01) for H342 and r=-0.38, (P=0.11) for PI. CONCLUSIONS: The increase in fluorescence values of membrane-ruptured H258 and unruptured H342-stained spermatozoa in boar AI doses can be associated with smaller litter size after AI. This finding indicates that the fluorescence properties of the sperm nucleus could be used to select for AI doses with greater fertilizing potential. PMID- 20015378 TI - A simulating analysis of the effects of increased joint stiffness on muscle loading in a thumb. AB - BACKGROUND: The development of osteoarthritis (OA) in the hand results in increased joint stiffness, which in turn affects the grip strength. The goal of the present study is to theoretically analyze the muscle forces in a thumb in response to the increased joint stiffness. METHODS: The thumb was modeled as a linkage system consisting of a trapezium, a metacarpal bone, a proximal and a distal phalanx. Nine muscles were included in the model: flexor pollicis longus (FPL), extensor pollicis longus (EPL), extensor pollicis brevis (EPB), abductor pollicis longus (APL), flexor pollicis brevis (FPB), abductor pollicis brevis (APB), the transverse head of the adductor pollicis (ADPt), the oblique head of the adductor pollicis (ADPo), and opponens pollicis (OPP). Numerical tests were performed using an inverse dynamic approach. The joints were prescribed to an angular motion at one degree-of-freedom (DOF) each time with all other DOFs of the joints being mechanically constrained, while the muscle forces in response to the joint motions were predicted. The normal joint stiffness was assumed to be 0.05, 0.10, and 0.15 N m/rad for interphalangeal (IP), metacarpophalangeal (MCP), and carpometacarpal (CMC) joint, respectively. The joint stiffness was assumed to increase by 50% and 100%, simulating the biomechanical consequences of OA. RESULTS: Our simulations indicated that the increase in joint stiffness induced substantial increases in muscle forces, especially in the EPL and FPL muscles in response to IP, MCP, or CMC extension/flexion motions. CONCLUSIONS: Because the strength of the muscles in the fingers is limited, the muscles will not be able to overcome joint resistance if joint stiffness is increased to its limit due to OA. This may contribute to the reduced range of motion typically seen in OA. PMID- 20015379 TI - Impact of genome assembly status on ChIP-Seq and ChIP-PET data mapping. AB - BACKGROUND: ChIP-Seq and ChIP-PET can potentially be used with any genome for genome wide profiling of protein-DNA interaction sites. Unfortunately, it is probable that most genome assemblies will never reach the quality of the human genome assembly. Therefore, it remains to be determined whether ChIP-Seq and ChIP PET are practicable with genome sequences other than a few (e.g. human and mouse). FINDINGS: Here, we used in silico simulations to assess the impact of completeness or fragmentation of genome assemblies on ChIP-Seq and ChIP-PET data mapping. CONCLUSIONS: Most currently published genome assemblies are suitable for mapping the short sequence tags produced by ChIP-Seq or ChIP-PET. PMID- 20015380 TI - Identification of novel aspartic proteases from Strongyloides ratti and characterisation of their evolutionary relationships, stage-specific expression and molecular structure. AB - BACKGROUND: Aspartic proteases are known to play an important role in the biology of nematode parasitism. This role is best characterised in blood-feeding nematodes, where they digest haemoglobin, but they are also likely to play important roles in the biology of nematode parasites that do not feed on blood. In the present work, we investigate the evolution and expression of aspartic proteases in Strongyloides ratti, which permits a unique comparison between parasitic and free-living adult forms within its life-cycle. RESULTS: We identified eight transcribed aspartic protease sequences and a further two genomic sequences and compared these to homologues in Caenorhabditis elegans and other nematode species. Phylogenetic analysis demonstrated a complex pattern of gene evolution, such that some S. ratti sequences had a one-to-one correspondence with orthologues of C. elegans but that lineage-specific expansions have occurred for other aspartic proteases in these two nematodes. These gene duplication events may have contributed to the adaptation of the two species to their different lifestyles. Among the set of S. ratti aspartic proteases were two closely-related isoforms that showed differential expression during different life stages: ASP-2A is highly expressed in parasitic females while ASP-2B is predominantly found in free-living adults. Molecular modelling of the ASP-2 isoforms reveals that their substrate specificities are likely to be very similar, but that ASP-2B is more electrostatically negative over its entire molecular surface than ASP-2A. This characteristic may be related to different pH values of the environments in which these two isoforms operate. CONCLUSIONS: We have demonstrated that S. ratti provides a powerful model to explore the genetic adaptations associated with parasitic versus free-living life-styles. We have discovered gene duplication of aspartic protease genes in Strongyloides and identified a pair of paralogues differentially expressed in either the parasitic or the free-living phase of the nematode life-cycle, consistent with an adaptive role for aspartic proteases in the evolution of nematode parasitism. PMID- 20015381 TI - RNA interference as a gene silencing therapy for mutant MYOC protein in primary open angle glaucoma. AB - BACKGROUND: Primary open-angle glaucoma (POAG) is the most common form of glaucoma which is an irreversible blind leading disease and lacks effective remedies. In recent years, POAG has been linked to the gene MYOC encoding myocilin that has been identified to harbor causal mutations. A variety of studies show that the mutant myocilin acts by gain of function. The mutant MYOC protein induces endoplasmic reticulum (ER) stress and the resultant unfolded protein response (UPR) induces apoptosis in the trabecular meshwork cells, which then leads to an increase in resistance to aqueous humor outflow, elevated intraocular pressure (IOP), and, ultimately, glaucoma. Culturing human trabecular meshwork (HTM) cells at a condition facilitating protein folding promotes secretion of mutant myocilin, normalizes cell morphology and reverses cell lethality. PRESENTATION OF THE HYPOTHESIS: We speculate that a complete elimination of mutant myocilin expression in trabecular meshwork cells is safe and that gives the possibility of avoiding the POAG phenotype. TESTING THE HYPOTHESIS: We propose RNA interference (RNAi) as a gene silencing therapy to eliminate the mutant myocilin proteins in the trabecular meshwork cells, either in a mutation-dependent or mutation-independent way due to the different engineering of the small interfering (si) RNA. IMPLICATIONS OF THE HYPOTHESIS: The RNAi strategy can reverse the pathological process of trabecular meshwork cells and thus treat the POAG caused by myocilin gene mutation. This strategy can also be applicable to many protein-misfolding diseases caused by gain-of-function mutant proteins. PMID- 20015382 TI - The protein tyrosine phosphatase receptor type R gene is an early and frequent target of silencing in human colorectal tumorigenesis. AB - BACKGROUND: Tumor development in the human colon is commonly accompanied by epigenetic changes, such as DNA methylation and chromatin modifications. These alterations result in significant, inheritable changes in gene expression that contribute to the selection of tumor cells with enhanced survival potential. RESULTS: A recent high-throughput gene expression analysis conducted by our group identified numerous genes whose transcription was markedly diminished in colorectal tumors. One of these, the protein-tyrosine phosphatase receptor type R (PTPRR) gene, was dramatically downregulated from the earliest stages of cellular transformation. Here, we show that levels of both major PTPRR transcript variants are markedly decreased (compared with normal mucosal levels) in precancerous and cancerous colorectal tumors, as well in colorectal cancer cell lines. The expression of the PTPRR-1 isoform was inactivated in colorectal cancer cells as a result of de novo CpG island methylation and enrichment of transcription repressive histone-tail marks, mainly H3K27me3. De novo methylation of the PTPRR 1 transcription start site was demonstrated in 29/36 (80%) colorectal adenomas, 42/44 (95%) colorectal adenocarcinomas, and 8/8 (100%) liver metastases associated with the latter tumors. CONCLUSIONS: Epigenetic downregulation of PTPRR seems to be an early alteration in colorectal cell transformation, which is maintained during the clonal selection associated with tumor progression. It may represent a preliminary step in the constitutive activation of the RAS/RAF/MAPK/ERK signalling, an effect that will later be consolidated by mutations in genes encoding key components of this pathway. PMID- 20015384 TI - Base composition, selection, and phylogenetic significance of indels in the recombination activating gene-1 in vertebrates. AB - BACKGROUND: The Recombination Activating Proteins, RAG1 and RAG2, play a crucial role in the immune response in vertebrates. Among the nuclear markers currently used for phylogenetic purposes, Rag1 has especially enjoyed enormous popularity, since it successfully contributed to elucidating the relationships among and within a large variety of vertebrate lineages. We here report on a comparative investigation of the genetic variation, base composition, presence of indels, and selection in Rag1 in different vertebrate lineages (Actinopterygii, Amphibia, Aves, Chondrichthyes, Crocodylia, Lepidosauria, Mammalia, and Testudines) through the analysis of 582 sequences obtained from Genbank. We also analyze possible differences between distinct parts of the gene with different type of protein functions. RESULTS: In the vertebrate lineages studied, Rag1 is over 3 kb long. We observed a high level of heterogeneity in base composition at the 3(rd )codon position in some of the studied vertebrate lineages and in some specific taxa. This result is also paralleled by taxonomic differences in the GC content at the same codon position. Moreover, positive selection occurs at some sites in Aves, Lepidosauria and Testudines. Indels, which are often used as phylogenetic characters, are more informative across vertebrates in the 5' than in the 3'-end of the gene. When the entire gene is considered, the use of indels as phylogenetic character only recovers one major vertebrate clade, the Actinopterygii. However, in numerous cases insertions or deletions are specific to a monophyletic group. CONCLUSIONS: Rag1 is a phylogenetic marker of undoubted quality. Our study points to the need of carrying out a preliminary investigation on the base composition and the possible existence of sites under selection of this gene within the groups studied to avoid misleading resolution. The gene shows highly heterogeneous base composition, which affects some taxa in particular and contains sites under positive selection in some vertebrate lineages in the 5'-end. The first part of the gene (5'-end) is more variable than the second (3'-end), and less affected by a heterogeneous base composition. However, in some vertebrate lineages the 5'-end of the gene is not yet widely used for phylogenetic studies. PMID- 20015383 TI - Sequence determinants of human microsatellite variability. AB - BACKGROUND: Microsatellite loci are frequently used in genomic studies of DNA sequence repeats and in population studies of genetic variability. To investigate the effect of sequence properties of microsatellites on their level of variability we have analyzed genotypes at 627 microsatellite loci in 1,048 worldwide individuals from the HGDP-CEPH cell line panel together with the DNA sequences of these microsatellites in the human RefSeq database. RESULTS: Calibrating PCR fragment lengths in individual genotypes by using the RefSeq sequence enabled us to infer repeat number in the HGDP-CEPH dataset and to calculate the mean number of repeats (as opposed to the mean PCR fragment length), under the assumption that differences in PCR fragment length reflect differences in the numbers of repeats in the embedded repeat sequences. We find the mean and maximum numbers of repeats across individuals to be positively correlated with heterozygosity. The size and composition of the repeat unit of a microsatellite are also important factors in predicting heterozygosity, with tetra-nucleotide repeat units high in G/C content leading to higher heterozygosity. Finally, we find that microsatellites containing more separate sets of repeated motifs generally have higher heterozygosity. CONCLUSIONS: These results suggest that sequence properties of microsatellites have a significant impact in determining the features of human microsatellite variability. PMID- 20015385 TI - Increased epithelial stem cell traits in advanced endometrial endometrioid carcinoma. AB - BACKGROUND: It has been recognized cancer cells acquire characters reminiscent of those of normal stem cells, and the degree of stem cell gene expression correlates with patient prognosis. Lgr5(+) or CD133(+) epithelial stem cells (EpiSCs) have recently been identified and these cells are susceptible to neoplastic transformation. It is unclear, however, whether genes enriched in EpiSCs also contribute in tumor malignancy. Endometrial endometrioid carcinoma (EEC) is a dominant type of the endometrial cancers and is still among the most common female cancers. Clinically endometrial carcinoma is classified into 4 FIGO stages by the degree of tumor invasion and metastasis, and the survival rate is low in patients with higher stages of tumors. Identifying genes shared between advanced tumors and stem cells will not only unmask the mechanisms of tumor malignancy but also provide novel therapeutic targets. RESULTS: To identify EpiSC genes in late (stages III-IV) EECs, a molecular signature distinguishing early (stages I-II) and late EECs was first identified to delineate late EECs at the genomics level. ERBB2 and CCR1 were genes activated in late EECs, while APBA2 (MINT2) and CDK inhibitor p16 tumor suppressors in early EECs. MAPK pathway was significantly up in late EECs, indicating drugs targeting this canonical pathway might be useful for treating advanced EECs. A six-gene mini-signature was further identified to differentiate early from advanced EECs in both the training and testing datasets. Advanced, invasive EECs possessed a clear EpiSC gene expression pattern, explaining partly why these tumors are more malignant. CONCLUSIONS: Our work provides new insights into the pathogenesis of EECs and reveals a previously unknown link between adult stem cells and the histopathological traits of EECs. Shared EpiSC genes in late EECs may contribute to the stem cell-like phenotypes shown by advanced tumors and hold the potential of being candidate therapeutic targets and novel prognosis biomarkers. PMID- 20015386 TI - Prediction of protein-protein interaction sites using an ensemble method. AB - BACKGROUND: Prediction of protein-protein interaction sites is one of the most challenging and intriguing problems in the field of computational biology. Although much progress has been achieved by using various machine learning methods and a variety of available features, the problem is still far from being solved. RESULTS: In this paper, an ensemble method is proposed, which combines bootstrap resampling technique, SVM-based fusion classifiers and weighted voting strategy, to overcome the imbalanced problem and effectively utilize a wide variety of features. We evaluate the ensemble classifier using a dataset extracted from 99 polypeptide chains with 10-fold cross validation, and get a AUC score of 0.86, with a sensitivity of 0.76 and a specificity of 0.78, which are better than that of the existing methods. To improve the usefulness of the proposed method, two special ensemble classifiers are designed to handle the cases of missing homologues and structural information respectively, and the performance is still encouraging. The robustness of the ensemble method is also evaluated by effectively classifying interaction sites from surface residues as well as from all residues in proteins. Moreover, we demonstrate the applicability of the proposed method to identify interaction sites from the non-structural proteins (NS) of the influenza A virus, which may be utilized as potential drug target sites. CONCLUSION: Our experimental results show that the ensemble classifiers are quite effective in predicting protein interaction sites. The Sub EnClassifiers with resampling technique can alleviate the imbalanced problem and the combination of Sub-EnClassifiers with a wide variety of feature groups can significantly improve prediction performance. PMID- 20015387 TI - Concerns about anti-angiogenic treatment in patients with glioblastoma multiforme. AB - BACKGROUND: The relevance of angiogenesis inhibition in the treatment of glioblastoma multiforme (GBM) should be considered in the unique context of malignant brain tumours. Although patients benefit greatly from reduced cerebral oedema and intracranial pressure, this important clinical improvement on its own may not be considered as an anti-tumour effect. DISCUSSION: GBM can be roughly separated into an angiogenic component, and an invasive or migratory component. Although this latter component seems inert to anti-angiogenic therapy, it is of major importance for disease progression and survival. We reviewed all relevant literature. Published data support that clinical symptoms are tempered by anti angiogenic treatment, but that tumour invasion continues. Unfortunately, current imaging modalities are affected by anti-angiogenic treatment too, making it even harder to define tumour margins. To illustrate this we present MRI, biopsy and autopsy specimens from bevacizumab-treated patients.Moreover, while treatment of other tumour types may be improved by combining chemotherapy with anti-angiogenic drugs, inhibiting angiogenesis in GBM may antagonise the efficacy of chemotherapeutic drugs by normalising the blood-brain barrier function. SUMMARY: Although angiogenesis inhibition is of considerable value for symptom reduction in GBM patients, lack of proof of a true anti-tumour effect raises concerns about the place of this type of therapy in the treatment of GBM. PMID- 20015388 TI - One nutritional symbiosis begat another: phylogenetic evidence that the ant tribe Camponotini acquired Blochmannia by tending sap-feeding insects. AB - BACKGROUND: Bacterial endosymbiosis has a recurring significance in the evolution of insects. An estimated 10-20% of insect species depend on bacterial associates for their nutrition and reproductive viability. Members of the ant tribe Camponotini, the focus of this study, possess a stable, intracellular bacterial mutualist. The bacterium, Blochmannia, was first discovered in Camponotus and has since been documented in a distinct subgenus of Camponotus, Colobopsis, and in the related genus Polyrhachis. However, the distribution of Blochmannia throughout the Camponotini remains in question. Documenting the true host range of this bacterial mutualist is an important first step toward understanding the various ecological contexts in which it has evolved, and toward identifying its closest bacterial relatives. In this study, we performed a molecular screen, based on PCR amplification of 16S rDNA, to identify bacterial associates of diverse Camponotini species. RESULTS: Phylogenetic analyses of 16S rDNA gave four important insights: (i) Blochmannia occurs in a broad range of Camponotini genera including Calomyrmex, Echinopla, and Opisthopsis, and did not occur in outgroups related to this tribe (e.g., Notostigma). This suggests that the mutualism originated in the ancestor of the tribe Camponotini. (ii) The known bacteriocyte associated symbionts of ants, in Formica, Plagiolepis, and the Camponotini, arose independently. (iii) Blochmannia is nestled within a diverse clade of endosymbionts of sap-feeding hemipteran insects, such as mealybugs, aphids, and psyllids. In our analyses, a group of secondary symbionts of mealybugs are the closest relatives of Blochmannia. (iv) Blochmannia has cospeciated with its known hosts, although deep divergences at the genus level remain uncertain. CONCLUSIONS: The Blochmannia mutualism occurs in Calomyrmex, Echinopla, and Opisthopsis, in addition to Camponotus, and probably originated in the ancestral lineage leading to the Camponotini. This significant expansion of its known host range implies that the mutualism is more ancient and ecologically diverse than previously documented. Blochmannia is most closely related to endosymbionts of sap-feeding hemipterans, which ants tend for their carbohydrate-rich honeydew. Based on phylogenetic results, we propose Camponotini might have originally acquired this bacterial mutualist through a nutritional symbiosis with other insects. PMID- 20015389 TI - In silico method for systematic analysis of feature importance in microRNA-mRNA interactions. AB - BACKGROUND: MicroRNA (miRNA), which is short non-coding RNA, plays a pivotal role in the regulation of many biological processes and affects the stability and/or translation of mRNA. Recently, machine learning algorithms were developed to predict potential miRNA targets. Most of these methods are robust but are not sensitive to redundant or irrelevant features. Despite their good performance, the relative importance of each feature is still unclear. With increasing experimental data becoming available, research interest has shifted from higher prediction performance to uncovering the mechanism of microRNA-mRNA interactions. RESULTS: Systematic analysis of sequence, structural and positional features was carried out for two different data sets. The dominant functional features were distinguished from uninformative features in single and hybrid feature sets. Models were developed using only statistically significant sequence, structural and positional features, resulting in area under the receiver operating curves (AUC) values of 0.919, 0.927 and 0.969 for one data set and of 0.926, 0.874 and 0.954 for another data set, respectively. Hybrid models were developed by combining various features and achieved AUC of 0.978 and 0.970 for two different data sets. Functional miRNA information is well reflected in these features, which are expected to be valuable in understanding the mechanism of microRNA-mRNA interactions and in designing experiments. CONCLUSIONS: Differing from previous approaches, this study focused on systematic analysis of all types of features. Statistically significant features were identified and used to construct models that yield similar accuracy to previous studies in a shorter computation time. PMID- 20015390 TI - Evidence of uneven selective pressure on different subsets of the conserved human genome; implications for the significance of intronic and intergenic DNA. AB - BACKGROUND: Human genetic variation produces the wide range of phenotypic differences that make us individual. However, little is known about the distribution of variation in the most conserved functional regions of the human genome. We examined whether different subsets of the conserved human genome have been subjected to similar levels of selective constraint within the human population. We used set theory and high performance computing to carry out an analysis of the density of Single Nucleotide Polymorphisms (SNPs) within the evolutionary conserved human genome, at three different selective stringencies, intersected with exonic, intronic and intergenic coordinates. RESULTS: We demonstrate that SNP density across the genome is significantly reduced in conserved human sequences. Unexpectedly, we further demonstrate that, despite being conserved to the same degree, SNP density differs significantly between conserved subsets. Thus, both the conserved exonic and intronic genomes contain a significantly reduced density of SNPs compared to the conserved intergenic component. Furthermore the intronic and exonic subsets contain almost identical densities of SNPs indicating that they have been constrained to the same degree. CONCLUSION: Our findings suggest the presence of a selective linkage between the exonic and intronic subsets and ascribes increased significance to the role of introns in human health. In addition, the identification of increased plasticity within the conserved intergenic subset suggests an important role for this subset in the adaptation and diversification of the human population. PMID- 20015391 TI - Documentation of body mass index and control of associated risk factors in a large primary care network. AB - BACKGROUND: Body mass index (BMI) will be a reportable health measure in the United States (US) through implementation of Healthcare Effectiveness Data and Information Set (HEDIS) guidelines. We evaluated current documentation of BMI, and documentation and control of associated risk factors by BMI category, based on electronic health records from a 12-clinic primary care network. METHODS: We conducted a cross-sectional analysis of 79,947 active network patients greater than 18 years of age seen between 7/05 - 12/06. We defined BMI category as normal weight (NW, 18-24.9 kg/m2), overweight (OW, 25-29.9), and obese (OB, >or= 30). We measured documentation (yes/no) and control (above/below) of the following three risk factors: blood pressure (BP) or = 4 degrees was calculated. RESULTS: Exact examinations of the hallux angle could be conducted on a total of 1,579 individual feet. Only 23.9% out of 1,579 feet presented a straight position of the great toe. The others were characterized by lateral deviations (valgus position) at different degrees, equalling 10 degrees or greater in 14.2% of the children's feet.88.8% of 808 children examined wore indoor footwear that was of insufficient length, and 69.4% of 812 children wore outdoor shoes that were too short. A significant relationship was observed between the lengthwise fit of the shoes and the hallux angle: the shorter the shoe, the higher the value of the hallux angle. The relative risk (odds ratio) of a lateral hallux deviation of > or = 4 degrees in children wearing shoes of insufficient length was significantly increased. CONCLUSIONS: There is a significant relationship between the hallux angle in children and footwear that is too short in length. The fact that the majority of the children examined were wearing shoes of insufficient length makes the issue particularly significant. Our results emphasize the importance of ensuring that children's footwear fits properly. PMID- 20015411 TI - Insecticide resistance in Anopheles gambiae: data from the first year of a multi country study highlight the extent of the problem. AB - BACKGROUND: Insecticide resistance in malaria vectors is a growing concern in many countries which requires immediate attention because of the limited chemical arsenal available for vector control. The current extent and distribution of this resistance in many parts of the continent is unknown and yet such information is essential for the planning of effective malaria control interventions. METHODS: In 2008, a network was established, with financial support from WHO/TDR, to investigate the extent of insecticide resistance in malaria vectors in five African countries. Here, the results of bioassays on Anopheles gambiae sensu lato from two rounds of monitoring from 12 sentinel sites in three of the partner countries are reported. RESULTS: Resistance is very heterogeneous even over relatively small distances. Furthermore, in some sites, large differences in mortality rates were observed during the course of the malaria transmission season. Using WHO diagnostic doses, all populations from Burkina Faso and Chad and two of the four populations from Sudan were classified as resistant to permethrin and/or deltamethrin. Very high frequencies of DDT resistance were found in urban areas in Burkina Faso and Sudan and in a cotton-growing district in Chad. In areas where both An. gambiae s.s. and Anopheles arabiensis were present, resistance was found in both species, although generally at a higher frequency in An gambiae s.s. Anopheles gambiae s.l. remains largely susceptible to the organophosphate fenitrothion and the carbamate bendiocarb in the majority of the sentinel sites with the exception of two sites in Burkina Faso. In the cotton-growing region of Soumousso in Burkina Faso, the vector population is resistant to all four classes of insecticide available for malaria control. CONCLUSIONS: Possible factors influencing the frequency of resistant individuals observed in the sentinel sites are discussed. The results of this study highlight the importance of standardized longitudinal insecticide resistance monitoring and the urgent need for studies to monitor the impact of this resistance on malaria vector control activities. PMID- 20015413 TI - [The scope of prenatal diagnostic testing for chromosomal aberrations: broad or narrow? Ethical considerations on the choice of tests]. AB - In the Netherlands prenatal diagnosis after screening for chromosomal abnormalities is done by karyotyping and is restricted to pregnant women with an increased risk of a child with trisomy 21, 18 or 13. However, karyotyping will detect a wider range of chromosomal abnormalities. Replacing karyotyping by rapid aneuploidy diagnosis (RAD) - a test with a more narrow scope - is currently under discussion. A possible drawback of RAD is that some rare but clinically relevant abnormalities may be missed. A possible advantage is that pregnant women will not be confronted with outcomes that the screening was not initially directed at. Each delineation of the scope of prenatal testing implies a normative choice that requires justification. Moral principles can be invoked for both narrowing down and further broadening of the scope of testing. PMID- 20015412 TI - Solution structure of the equine infectious anemia virus p9 protein: a rationalization of its different ALIX binding requirements compared to the analogous HIV-p6 protein. AB - BACKGROUND: The equine infection anemia virus (EIAV) p9 Gag protein contains the late (L-) domain required for efficient virus release of nascent virions from the cell membrane of infected cell. RESULTS: In the present study the p9 protein and N- and C-terminal fragments (residues 1-21 and 22-51, respectively) were chemically synthesized and used for structural analyses. Circular dichroism and 1H-NMR spectroscopy provide the first molecular insight into the secondary structure and folding of this 51-amino acid protein under different solution conditions. Qualitative 1H-chemical shift and NOE data indicate that in a pure aqueous environment p9 favors an unstructured state. In its most structured state under hydrophobic conditions, p9 adopts a stable helical structure within the C terminus. Quantitative NOE data further revealed that this alpha-helix extends from Ser-27 to Ser-48, while the N-terminal residues remain unstructured. The structural elements identified for p9 differ substantially from that of the functional homologous HIV-1 p6 protein. CONCLUSIONS: These structural differences are discussed in the context of the different types of L-domains regulating distinct cellular pathways in virus budding. EIAV p9 mediates virus release by recruiting the ALG2-interacting protein X (ALIX) via the YPDL-motif to the site of virus budding, the counterpart of the YPXnL-motif found in p6. However, p6 contains an additional PTAP L-domain that promotes HIV-1 release by binding to the tumor susceptibility gene 101 (Tsg101). The notion that structures found in p9 differ form that of p6 further support the idea that different mechanisms regulate binding of ALIX to primary versus secondary L-domains types. PMID- 20015414 TI - [Missed malignant melanoma: legal considerations]. AB - Malignant melanoma may be missed both clinically and histopathologically. The appearance of metastases after local excision of a skin lesion which was not recognized as melanoma indicates a diagnostic error. In a lawsuit three things have to be proven: that the doctor has behaved negligently, that the patient has been damaged and that there is a causal relation between the two. After local excision of a lesion that subsequently proves to be a metastasized melanoma, no damage ('loss of chance') occurs because neither local surgery nor adjuvant therapy influences disease-free survival. These particularities are of importance in legal judgments. PMID- 20015415 TI - [Frailty in the elderly]. AB - Older people differ in their level of multimorbidity, functional dependence and need for assistance. Frailty is a recently constructed syndromatic entity used to characterize vulnerable older people with an increased risk of functional decline, institutionalization and death. This is illustrated by the case history of a 97-year-old woman. In addition to somatic multimorbidity, she suffered from fatigue, weight loss, diminished strength and mood, fear of falling and a decreased gait velocity, without a clear link to a disease. The term 'frailty' is useful in such cases and its application is quickly expanding. Frailty highlights the need to individualize and integrate guidelines for treatment, and to prevent adverse outcomes by choosing health care interventions fit for such frail elderly. However, in practice the frailty syndrome is defined by diverse sets of criteria. As frailty is used to predict different adverse outcomes, a single definition is not possible. Awareness of the advantages and pitfalls of the frailty concept may lead to fruitful clinical application. PMID- 20015417 TI - [Life expectancy increasing, but gap between people of higher and lower educational level remains]. AB - OBJECTIVE: To determine if since 1997 there has been a change in the gap in life expectancy between those of higher and lower educational level. DESIGN: Statistical analysis of register and survey data. METHOD: Linkage of data from municipal population registers and the Labour Force Survey of Statistics Netherlands. RESULTS: Life expectancy of both men and women was positively correlated with level of education. The life expectancy of Dutch men increased by about two years in the period 1997-2005, irrespective of level of education. Women gained about one year, but life expectancy among those of both lower and higher levels of education stagnated. During the study period the gap in life expectancy between these educational extremes remained almost constant at about 5 to 6 years. CONCLUSIONS: Our findings point to both favourable and unfavourable trends. All population groups seemed to have profited from a recent relatively strong increase in life expectancy which is clearly a positive development. Less favourable was the fact that the gap between those of lower and higher educational level had not decreased. PMID- 20015418 TI - [New understanding of the onset of atherosclerosis--angiogenesis and hypoxia play a crucial role]. AB - Clinical signs of atherosclerosis, such as heart attack and stroke, are often caused by rupture of the cap of an atherosclerotic plaque, with thrombus formation as a consequence. The risk of rupture depends on the formation of microvessels (angiogenesis) in the plaque. The fragility of the microvascular endothelium causes hyperpermeability, which leads to intraplaque haemorrhage. Angiogenesis is stimulated by hypoxia, oxidative stress and the production of hypoxia-inducible transcription factors. Hypoxia is primarily caused by increased oxygen consumption of inflammatory cells, while plaque thickness, which reduces oxygen diffusion, contributes to a limited extent. A vicious circle of hypoxia, (incomplete) angiogenesis and inflammation occurs deep in the plaque, which enhances plaque growth and the risk of plaque rupture. By non-invasive imaging of plaque hypoxia and angiogenesis, plaques at risk of rupture may be identified. Therapeutic interventions for plaque angiogenesis and hypoxia require further investigation. PMID- 20015419 TI - [Phyllodes tumour: a rare, rapidly growing breast tumour]. AB - A 40-year-old woman presented at the breast outpatient clinic with a giant tumour of her left breast. The size, rapid growth and radiological characteristics of the lesion led us to suspect a phyllodes tumour. A histological examination of a needle biopsy confirmed this diagnosis. An additional CT scan revealed no signs of metastases. We performed a mastectomy during which a tumour measuring 48 x 33 x 25 cm was resected. Histological examination revealed a borderline phyllodes tumour. Phyllodes tumours are rare fibroepithelial neoplasms of the breast and pre-operatively these are often difficult to differentiate from fibroadenomas. Phyllodes tumours have a variable clinical course with the ability to metastasize and a propensity to recur locally. Complete excision with wide margins is essential to prevent local recurrence. In our case, the surgical margins were limited and our patient was therefore treated with postoperative radiation therapy. PMID- 20015420 TI - [A man with a swelling of his navel]. AB - A patient with liver cirrhosis and portal hypertension demonstrated the rerouting of blood through the veins of the abdominal wall. PMID- 20015421 TI - Public Health Nutrition for this decade. PMID- 20015423 TI - Running away with the facts on food and fatness. PMID- 20015424 TI - Letter to the editor. PMID- 20015425 TI - Systematic review of the effectiveness and cost-effectiveness of weight management schemes for the under fives: a short report. AB - OBJECTIVE: To search for, review and synthesise studies of the effectiveness and cost-effectiveness of weight management schemes for the under fives. DATA SOURCES: MEDLINE [Ovid], MEDLINE In-Process [Ovid], EMBASE [Ovid], CAB [Ovid], Health Management Information Consortium [Ovid], The Cochrane Database of Systematic Reviews, Cochrane Register of Controlled Trials, Science Citation Index Expanded [Web of Science], Conference Proceedings Citation Index [The Web of Science], Database of Abstract Reviews [CRD; Centre for Reviews and Dissemination], HTA [CRD], PsycINFO [Ebsco], NHS CRD. These databases were searched from 1990 to February 2009. Supplementary internet searches were additionally conducted. REVIEW METHODS: Relevant clinical effectiveness studies were identified in two stages. Titles and abstracts returned by the search strategy were examined independently by three researchers and screened for possible inclusion. Disagreements were resolved by discussion. Full texts of the identified studies were obtained. Three researchers examined these independently for inclusion or exclusion, and disagreements were again resolved by discussion. RESULTS: One of the randomised controlled trials (RCTs) was from the UK. It measured the effects of a physical activity intervention for children in nurseries combined with home-based health education for their parents; this was compared to usual care. The main outcome measure was body mass index (BMI); secondary measures were weight and physical activity. At the 12-month follow-up, no statistically significant differences were found between the groups on any measure. However, a trend, favouring the intervention, was found for BMI and weight. The other two RCTs were from the USA. The larger trial investigated the effects of a combined preschool and home intervention in African American and Latino communities. Nutrition education and physical activity programmes were aimed at under fives in preschool. The home component consisted of related health education and homework for the parents, who received a small financial reward on completion. The 1- and 2-year results for the African American sites showed a significantly slower rate of increase in BMI than for results at baseline, for the intervention group than for the control group. However, in the Latino communities no such differences were found. The second US trial was a much smaller home-based parental education programme in Native American communities in the USA and Canada. The intervention consisted of a parental skills course for parents to improve their children's diet and physical activity. This was compared with a course providing skills to improve child behaviour. Follow-up was at 16 weeks and showed no significant differences between groups in BMI. CONCLUSIONS: No controlled trials addressing the issue of treating obesity or evidence of cost effectiveness studies in the under fives' population were found. From the three prevention studies, apart from the larger US trial, the interventions showed no statistically significant differences in BMI and weight between the intervention and control groups (although there was some evidence of positive trends for BMI and weight). It should also be noted that these conclusions are based on only three dissimilar studies, thereby making the drawing of firm conclusions difficult. Research is urgently needed in further well-designed UK-based RCTs of weight management schemes aimed at the prevention of obesity, that combine with cost-effectiveness studies targeted at preschool children with long-term follow up. PMID- 20015426 TI - [Current problems and countermeasures in treatment of complex anal fistula]. AB - The high recurrent rate and insufficient protection of anal function in treatment of anal fistula were reviewed in this article. To solve the problems in the treatment of complex fistula-in-ano including ignorance of preoperative assessment and lack of consciousness of preserving anal function, the authors raised the countermeasures to carry out the "gold standard" of preoperative assessment and the system of medical delivery, form the new principle of incision of anal sphincters, collect the high-level evidence-based medicine proof, improve the healing rate and significantly reduce the postoperative continence disturbance, so as to develop a new guideline for the treatment of fistula-in ano. PMID- 20015427 TI - [Progress in surgical treatment of perianal abscess]. AB - There were more and more patients suffered from perianal abscess. The patient's condition will become even worse if the patients cannot be treated properly. Selecting proper operation method according to special condition is the key point of increasing curative effect, decreasing suffering and relapse rate and promoting quality of life of patients. The authors summarized operation methods, including abscess incision drainage, radical incision and thread-drawing, and staging incision and thread-drawing, etc. in treating perianal abscess in clinic. PMID- 20015428 TI - [Recognition and thoughts for diagnosis and treatment of chronic prostatitis in integrated traditional Chinese and Western medicine]. AB - The etiology, pathogenesis, and diagnostic criteria of chronic prostatitis were reviewed in this article. Based on clinical practice, the authors systematically discussed the thoughts and methods for the treatment of chronic prostatitis by integrated traditional Chinese and Western medicine. Meanwhile, advice on disputed problems in clinical study of prostatits were given, such as curative effect estimation value of the number of leukocytes in expressed prostatic secretion (EPS) and bacterial culture in EPS, the opportunity and treatment course of antibiotics, National Institutes of Health chronic prostatitis symptom index, classification of syndromes of traditional Chinese medicine (TCM), TCM symptom score, and clinical study period. PMID- 20015429 TI - [Design characteristics of clinical surgery trial based on treatment program of tunnel thread-drawing method for anal fistula: a prospective randomized controlled multicenter trial]. AB - BACKGROUND: Basic principles of clinical trials of nonpharmacologic treatment are similar to those of pharmacologic treatment, but its some special characteristics should be discussed. OBJECTIVE: To explore the design characteristics of clinical surgery trial through the example of tunnel thread-drawing therapy for simple anal fistula. DESIGN, SETTING, PARTICIPANTS AND INTERVENTIONS: The clinical trial was designed as a prospective, controlled, randomized multicenter trial. The patients came from Longhua Hospital, Yueyang Hospital of Integrated Traditional Chinese and Western medicine, and Affiliated Hospital of Jianxi University of Traditional Chinese Medicine. Stratified random was performed according to the subtype of anal fistula including low and high anal fistula. Full analysis set was applied to analyze the baseline data, and per protocol set was used in efficacy and safety analysis. The intervention was tunnel thread-drawing method. Classical therapy of thread-drawing method was employed as positive control. MAIN OUTCOME MEASURES: The primary outcomes were course of recovery and cure rate. Quality of life score was used as secondary outcome and anal maximum constriction pressure was considered as safety outcome. RESULTS: Optimal efficiency testing method was used to estimate sample size. A total of 244 subjects were recruited and 236 subjects completed the trial. There was no significant difference in the cure rate of the low and high anal fistula between the treatment group and control group. There was a significant difference in the course of recovery in the patients with low and high anal fistula between treatment group and control group (P<0.01). To the patients with low anal fistula, the course of recovery in the treatment group was (22.26 + or - 8.67) d, and the course of recovery in the control group was (31.41+ or - 11.39) d. To the patients with high anal fistula, the course of recovery in the treatment group was (24.73 + or - 8.15) d, and the course of recovery in the control group was (32.20 + or - 12.60) d. There was no significant difference in the scores of quality of life in the patients with low anal fistula between the treatment group and control group (P>0.05). The trial showed that the tunnel thread-drawing therapy was significantly more effective than classical method for improving the anal sphincter function and patient satisfaction with treatment in the patients with high anal fistula. However the other items of quality of life in the two groups did not show significant difference. There was no any adverse event report in each group. There was no significant difference in the anal maximum constriction pressure in the treatment group before and after the operation. CONCLUSION: The trial shows that the tunnel thread-drawing therapy for simple anal fistula can shorten the course of recovery and improve the patients' quality of life. The training about surgical intervention and clinical implementation program is important in clinical surgery trial. Blind is absolutely difficult to implement and placebo cannot be used in this kind of trial. PMID- 20015430 TI - [Tunnel thread-drawing therapy in treating patients with horseshoe-shaped low position perianal abscess: a randomized controlled trial]. AB - BACKGROUND: Tunnel thread-drawing therapy is an effective treatment for complex anal fistula, and this therapy may be used for horseshoe-shaped low-position perianal abscess. OBJECTIVE: To analyze the therapeutic effects of tunnel thread drawing therapy in treatment of horseshoe-shaped low-position perianal abscess. DESIGN, SETTING, PARTICIPANTS AND INTERVENTIONS: A total of 68 patients with horseshoe-shaped low-position perianal abscess were from Department of Anorectal Surgery, Shanghai Tianshan Traditional Chinese Medicine Hospital, and were randomly divided into treatment group and control group. There were 34 cases in each group. The patients in the treatment group received thread-drawing therapy and the patients in the control group received multiple incisions and drainage. MAIN OUTCOME MEASURES: Lasting time of postoperative pain, healing time of incision, average area of the wound scar, anal function and postoperative recurrence were observed in the two groups. Late postoperative follow-up was from half a year to three years. RESULTS: The lasting time of postoperative pain and healing time of incision in the treatment group were shortened as compared with those in the control group (P<0.01). There was no significant difference in postoperative recurrence rate between the two groups (P>0.05). CONCLUSION: There is a significant curative effect of tunnel thread-drawing therapy in treatment of horseshoe-shaped low-position perianal abscess. PMID- 20015431 TI - [Yunchang Capsule in treatment of functional constipation: a randomized, double blinded controlled, multicenter trial]. AB - BACKGROUND: Although there are some Chinese herbal medicines in treatment of constipation, but no multi-center randomized controlled trials have been carried out to prove their effectiveness. OBJECTIVE: To evaluate the safety and efficacy of Yunchang Capsule in treatment of functional constipation with deficiency of both qi and yin and internal accumulation of poisonous pathogenic factors syndrome, and to explore the clinical dosage. DESIGN, SETTING, PARTICIPANTS AND INTERVENTIONS: A randomized, double-blinded controlled, multicenter trial was conducted. A total of 240 patients with functional constipation from West China Hospital of Sichuan University, Affiliated Hospital of Chengdu University of Traditional Chinese Medicine, Affiliated Hospital of Jiangxi University of Traditional Chinese Medicine, the First Affiliated Hospital of Tianjing University of Traditional Chinese Medicine and Fujian Academy of Traditional Chinese Medicine were randomly divided into three groups: low dose group (80 cases), high dose group (80 cases) and control group (80 cases). Patients in the low dose group were treated with two pills (0.35 g/pill) of Yunchang Capsule and one pill of Yunchang Capsule simulant for three times daily; patients in the high dose group were treated with three pills (0.35 g/pill) of Yunchang Capsule for three times daily; and patients in the control group were treated with three pills (0.35 g/pill) of Biantong Capsule for three times daily. The therapeutic course was 14 days. MAIN OUTCOME MEASURES: Clinical symptoms, syndromes, and adverse effects were observed before and after the treatment, and blood, urine and stool tests, hepatorenal function and electrocardiogram were also examined. RESULTS: Two cases were excluded, eleven cases were lost to follow-up, and there were 234 patients entered to intention-to-treat (ITT) analysis. After the treatment, the therapeutic effects were calculated by full analysis set (FAS) and per-protocol population set (PPS) analysis respectively. The effects on functional constipation in FAS showed the response rates in the low dose, high dose and control groups were 86.25% (69/80), 82.90% (63/76), and 70.52% (55/78) respectively, and PPS analysis showed the response rates were 85.71% (66/77), 83.56% (61/73), and 70.13% (54/77) respectively. There were no significant differences among the three groups (P>0.05). The effects on traditional Chinese medicine syndrome in FAS showed the response rates in the low dose, high dose and control groups were 78.75% (63/80), 69.74% (53/76), and 67.95% (53/78) respectively, and PPS analysis showed the response rates were 77.92% (60/77), 69.87%(51/73), and 67.53% (52/77) respectively. There were also no significant differences among the three groups (P>0.05). No severe adverse events were observed. CONCLUSION: Both low dose and high dose of Yunchang Capsule are effective and safe in treatment of functional constipation with deficiency of both qi and yin and internal accumulation of poisonous pathogenic factors syndrome. PMID- 20015432 TI - [Kneading and dispersing manipulation in treatment of early-stage acute mastitis: a randomized controlled trial]. AB - BACKGROUND: Acute mastitis is an acute infectious disease of breast. Antibiotic treatment is often unable to obtain a good effect, and we should actively look for a safe and effective non-drug therapy. OBJECTIVE: To validate the clinical efficacy of kneading and dispersing manipulation in treatment of early-stage acute mastitis. DESIGN, SETTING, PARTICIPANTS AND INTERVENTIONS: According to the multicenter randomized controlled trial design, 198 cases of acute mastitis from Yueyang Hospital of Integrated Traditional Chinese Medicine, Longhua Hospital, and Shanghai Yangpu Maternity and Child Health Hospital were randomly divided into treatment group and control group. There were 99 cases in each group. Patients in the treatment group were only treated with manipulation, and cefradine was orally administered to patients in the control group. MAIN OUTCOME MEASURES: The local breast lump size, clinical symptoms and the adverse reactions in the two groups were observed before and after the treatment. RESULTS: The total response rates in the treatment and control group were 95.92% (94/98) and 80% (76/95) respectively. There was a significant difference in the total response rate between the two groups (P<0.05). There were significant differences in the score of breast lump size, and the score of signs and symptoms between the two groups (P<0.05). CONCLUSION: Kneading and dispersing manipulation has certain effects on early-stage acute mastitis, and the therapy is safe and repeatable. PMID- 20015433 TI - [Influence of integrated traditional Chinese and Western medicine therapy on serum resistin levels in patients with severe acute pancreatitis: a randomized controlled trial]. AB - BACKGROUND: Resistin level is high in patients with severe acute pancreatitis (SAP), and resistin is expected to be a new marker for evaluating the severity of acute pancreatitis. OBJECTIVE: To explore the influence of integrated traditional Chinese and Western medicine therapy on serum resistin levels in SAP patients. DESIGN, SETTING, PARTICIPANTS AND INTERVENTIONS: Twenty-eight SAP patients meeting inclusion criteria from Department of Integrated Traditional Chinese and Western Medicine, West China Hospital, Sichuan University were included, and the patients were randomly divided into treatment group and placebo group. There were 13 patients in the treatment group and 15 patients in the placebo group. Patients in the treatment group were given traditional Chinese herbal medicine in addition to the conventional treatment. Patients in the placebo group were given placebo in addition to the conventional treatment. MAIN OUTCOME MEASURES: The serum resistin levels on admission, and days 1, 3, 5, and 7 after the admission were detected. RESULTS: The serum resistin levels on admission in all the patients were higher than normal level, and there was no significant difference between the two groups (P>0.05). On days 1, 3, 5, and 7 after admission, the resistin levels in the treatment group were (3.29 + or - 1.66) microg/L, (3.71 + or - 1.05) microg/L, (3.08 + or - 1.47) microg/L and (3.62 + or - 1.67) microg/L, and in the control group (5.16 + or - 1.93) microg/L, (5.07 + or - 1.53) microg/L, (4.88 + or - 1.47) microg/L and (5.12 + or - 1.48) microg/L, respectively. The resistin levels were lower in the treatment group than in the control group (P<0.05). CONCLUSION: Serum resistin level in SAP patients can be decreased by integrated traditional Chinese medicine and Western medicine therapy. PMID- 20015434 TI - [Clinical study of syndrome differentiation standards for chronic skin ulcer of lower limbs]. AB - OBJECTIVE: To establish evidence and standards for syndrome differentiation of patients with chronic skin ulcer of lower limbs, which can be used to supervise and further improve the clinical treatment. The evidence and standards would also lay a foundation for the integrative evaluation system of clinical effects. METHODS: This is a prospective study of clinical case collection and analysis. The syndrome factors of chronic skin ulcer of lower limbs were collected from clinical cases, and then the essential factors of syndromes were chosen to make cluster analysis and principal component analysis. After syndrome classification, specialist consultation was performed. Forecasting syndrome models were established on the basis of the statistical analysis and the specialist consultation. RESULTS: The syndrome factors in 338 cases of chronic skin ulcer of lower limbs were clustered into four syndrome categories including deficiency of both qi and blood, spleen deficiency with dampness encumbrance, qi deficiency with blood stasis and blood stagnation with dampness-heat. The deficiency of both qi and blood, and qi deficiency with blood stasis were merged into qi deficiency with blood stasis by expert revision. The primary and secondary symptoms were identified by principal component analysis and average ranking determined by experts. The qi deficiency with blood stasis, spleen deficiency with dampness encumbrance, and blood stagnation with dampness-heat could reflect the clinical syndrome characteristics in patients with chronic skin ulcer of lower limbs. CONCLUSION: Chronic skin ulcer of lower limbs can be differentiated in three syndrome types, and differentiation of local symptoms of the ulcer is very important in syndrome differentiation of chronic skin ulcer of lower limbs. PMID- 20015435 TI - [Effects and action mechanisms of Buyang Huanwu Decoction in wound healing of chronic skin ulcers of rats]. AB - OBJECTIVE: To explore the effects and action mechanisms of Buyang Huanwu Decoction, a compound traditional Chinese herbal medicine, in wound healing of chronic skin ulcers of rats. METHODS: A total of 54 male SD rats with back full thickness skin lesion were used in this study. Except for the normal control group, the rats were injected with hydrocortisone for inducing chronic skin ulcers, and were randomly divided into untreated group and Buyang Huanwu Decoction group. The rate and the time of wound healing were observed. Immunohistochemical method and image analytical method were used to test the expression of vascular endothelial growth factor (VEGF) and microvessel count (MVC) in granulation tissue of chronic skin ulcers in the rats. RESULTS: The rate of wound healing was significantly lowered in the untreated group as compared with the normal control group (P<0.01, P<0.05), and the time of wound healing was obviously longer too (P<0.01). In the Buyang Huanwu Decoction group, the rate of wound healing was significantly higher than that in the untreated group (P<0.01), and the time of wound healing was obviously shorter than the latter's (P<0.01). In the untreated group, the expression of VEGF and MVC were significantly lower than those in the normal control group (P<0.01); the expression of VEGF and MVC were obviously increased in the Buyang Huanwu Decoction group as compared with those in the untreated group (P<0.01). CONCLUSION: Buyang Huanwu Decoction has the effect to promote the wound healing for chronic skin ulcers of rats by regulating the expression of VEGF in granulation tissue of chronic skin ulcers. PMID- 20015436 TI - [Effects of oral and external application of recipes for promoting granulation in rats with chronic skin ulcers]. AB - OBJECTIVE: To investigate the efficacy of oral, external and oral plus external application of recipes for promoting granulation on chronic skin ulcers in rats. METHODS: Sixty SD male rats were randomly divided into normal group, untreated group, oral group, external group and combined group (n=12). Chronic skin ulcer was induced by hydrocortisone injection and back skin incision. After oral administration of Fuzheng Huyu Recipe or (and) external application of Baiyu ointment, the compound traditional Chinese medicines for wound healing, healing time and healing rate of the ulcers were observed. RESULTS: The healing rate was lower and the healing time was longer in the untreated group than in the normal group (P<0.01). The healing rates of the oral group and the combined group were higher than those of the untreated group at all observation points (P<0.05), and the healing rate of the external group was higher on the 3rd and 14th day after model establishment (P<0.05). On the 7th and 10th day after model establishment, the healing rates of the oral group and the combined group were higher than those of the external group (P<0.05), and the healing time of the combined group was shorter than that of the oral group and external group (P<0.05). CONCLUSION: Both orally and externally administered wound healing recipes can promote the restoration of chronic refractory ulcers; the effect of combined administration is superior to single administration. Oral, external and combined application can all improve the wound healing in inflammatory phase. Oral and combined application can obviously improve the wound healing in granulation phase and epithelial phase. PMID- 20015437 TI - [Effects of mild moxibustion on angiogenesis and microcirculation in wound repair after operation of anal fistula in rats]. AB - OBJECTIVE: To observe the effects of mild moxibustion on angiogenesis and microcirculation in wound repair after operation of anal fistula, and to explore the mechanism of action. METHODS: Seventy-two SD male rats were made to bear an "acute, open, bleeding, infected" wound surface and were divided randomly into mild moxibustion group, microwave group and control group (24 rats per group). The wound surfaces of the treatment groups were treated for 15 min with mild moxibustion or microwave medical instrument from the first day after operation, and the control group was not given any treatment. At the first, third, seventh and fourteenth days after operation (1, 3, 7 and 14 days of treatment), six rats in each group were sacrificed after examining the blood flow of the wound surface, and the samples of granulation tissues were collected. The numbers of CD34 and positive expression of vascular endothelial growth factor (VEGF) in granulation tissues were detected. RESULTS: In the early and middle stages of wound healing, mild moxibustion could obviously increase the expression of VEGF, the number of CD34, and the blood flow of the wound surface. And the expression of VEGF, the number of CD34, and the blood flow of the wound surface began to increase at the third day, and reached the peak level at the seventh day, and then began to decrease. In the late stage of wound healing (day 14), mild moxibustion could decrease the expression of VEGF, the number of CD34, and decrease the blood flow of the wound surface. There were significant differences between the mild moxibustion group and the control group in the blood flow of the wound surface, the expression of VEGF and the numbers of CD34 in granulation tissues (P<0.05) after treatment, and except the number of CD34 at 14-day treatment, there were also differences between the mild moxibustion group and the microwave group. CONCLUSION: Mild moxibustion has better regulative actions on blood flow of the wound surface and the number of CD34 and positive expression of VEGF in granulation tissues, and can regulate microcirculation in wound surface and promote the wound healing. PMID- 20015438 TI - [Effects of Yanggan Lidan Granule on insulin resistance in guinea pigs with induced cholesterol gallstones]. AB - OBJECTIVE: To observe the effects of Yanggan Lidan Granule (YGLDG), a compound traditional Chinese herbal medicine, on insulin resistance in guinea pigs with induced cholesterol gallstones. METHODS: Eighty guinea pigs were randomly divided into normal control group, untreated group, YGLDG group and ursodeoxycholic acid (UDCA) group, with 20 guinea pigs in each group. Except the normal control group, gallstones were induced by high-cholesterol diet in the guinea pigs. The guinea pigs in the normal control group and the untreated group were administered with normal saline. UDCA and YGLDG were given to the guinea pigs in the corresponding groups for seven weeks. Eight guinea pigs of each group were used to measure the glucose infusion rate (GIR) by using hyperinsulinemic-euglycemic clamp technique. At the end the guinea pigs were killed and their gallstone formation was observed. RESULTS: The gallstones in guinea pigs were identified as cholesterol stones by qualitative analysis through infrared spectrum. The incidence rate of cholelithiasis of the untreated group was 82.35% . The GIR of guinea pigs in the untreated group was obviously lowered down as compared with the normal control group. Compared with the untreated group, the GIRs of the YGLDG group and the UDCA group were obviously increased, especially in the YGLDG group. CONCLUSION: YGLDG may improve insulin resistance in guinea pigs with cholesterol gallstones by elevating GIR obviously. PMID- 20015439 TI - [Effects of Qingshi Cream on chronic dermatitis-eczema in mice]. AB - OBJECTIVE: To explore the effects of Qingshi Cream, a compound traditional Chinese herbal medicine, on chronic dermatitis-eczema in mice induced by 2,4 dinitrochlorobenzene (DNCB). METHODS: Thirty BALB/C mice were randomly divided into vaseline group, 0.1% mometasone furoate cream group and Qingshi Cream group. Right ears of BALB/C mice were repeatedly challenged with 0.1% DNCB every three days for five times and previously sensitized with 7% DNCB to induce chronic dermatitis-eczema. Mice in different groups were applied Qingshi Cream, 0.1% mometasone furoate cream and vaseline respectively after each challenge. Weight difference of two ears, pathological change of right ear and dermal inflammatory cell number were used to assess the effects of the drugs. RESULTS: After the 5th challenge, weight differences of two ears in the 0.1% mometasone furoate cream group and the Qingshi Cream group were significantly decreased as compared with that in the vaseline group. Changes such as ear swelling thickening and cellular infiltration in dermis were observed, and these features seemed to be more significant in the vaseline group as compared with the 0.1% mometasone furoate cream group and the Qingshi Cream group. CONCLUSION: Qingshi Cream has an obvious effect in treatment of chronic dermatitis-eczema in mice. PMID- 20015440 TI - [Effects of emodin on cell apoptosis of intestinal mucosa and serum leptin in rats with severe acute pancreatitis]. AB - OBJECTIVE: To explore the mechanisms of emodin in protecting intestinal mucosal barrier in rat with severe acute pancreatitis (SAP). METHODS: Sixty SD rats were randomly divided into three groups: sham-operation group, untreated group, and emodin group. SAP in rats of the untreated group and the emodin group was induced by retrograde pumping of 3.0% sodium cholate to the common bile duct. Specimens were obtained 24 hours after the severe acute pancreatitis was induced. Serum level of leptin, serum activity of amylase and plasma content of endotoxin were measured. Ileum mucosa from ileocecal junction was observed by light microscopy and electron microscopy to measure pathological and ultrastructural changes. Apoptosis of ileum mucosal cells was detected by terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling method, and expression of Bax in ileum mucosal cells was measured by immunohistochemical method. RESULTS: Compared with the sham-operation group, there was significant increase in the levels of leptin, endotoxin, the activity of amylase, apoptosis index and Bax expression in the untreated group (P<0.01). Compared with the untreated group, the level of endotoxin, apoptotic index and Bax expression level in the emodin group were significantly reduced (P<0.01) and the leptin level was increased (P<0.05). More severe pathological changes appeared in the untreated group than in the sham operation group under the light and electron microscopes; meanwhile less severe damage was observed in the emodin group as compared with the untreated group. CONCLUSION: Emodin can inhibit the apoptosis of intestinal mucosa cells and up regulate the serum leptin content to protect the intestina1 barrier function and prevent the translocation of bacteria and endotoxin. PMID- 20015441 TI - [Effects of Astragalus injection and its ingredients on proliferation and Akt phosphorylation of breast cancer cell lines]. AB - OBJECTIVE: To observe the effects of Astragalus injection, astragaloside IV and formononetin on proliferation and Akt phosphorylation of basal-like human breast carcinoma cell lines MDA-MB-468 and MDA-MB-231. METHODS: The effects of different concentrations of Astragalus injection, astragaloside IV and formononetin on proliferation of breast cancer cell lines were assayed by methyl thiazolyl tetrazolium (MTT) assay, and their effects on phospho-Akt were assayed by in-cell Western blot method. RESULTS: The results of the MTT assay showed that the best concentrations of Astragalus injection, astragaloside IV, formononetin and astragaloside IV plus formononetin were 1 g/mL, 80 microg/mL, 40 microg/mL and 10 microg/mL plus 40 microg/mL respectively. After 1- or 2-day culture, Astragalus injection, astragaloside IV, formononetin and astragaloside IV plus formononetin decreased the expressions of p-Akt (Thr 308) and p-Akt (Ser 473) in MDA-MB-468 cells. Formononetin and astragaloside IV plus formononetin down-regulated the expression of p-Akt (Thr 308) protein in MDA-MB-231 cells after 1- and 2-day culture, but had no effects on the expression of p-Akt (Ser 473) protein in MDA MB-231 cells. CONCLUSION: Astragalus injection, astragaloside IV and formononetin can inhibit proliferation of breast cancer cell lines MDA-MB-468 and MDA-MB-231, and the antiproliferation effects vary according to their concentrations. And the antiproliferation mechanisms may be related to their down-regulation effects on Akt phosphorylation. PMID- 20015442 TI - [Segmenting jugged dissection and ligation combined with additional ligation in treating circular mixed hemorrhoids: a report of 63 cases]. PMID- 20015443 TI - [Research advances in the mechanisms of antitumor activity of Epimedium brevicornum Maxim. and its effective components]. PMID- 20015444 TI - [Statistical analysis for data of multidimensional contingency table with SAS software package (part two)]. PMID- 20015445 TI - Comparative study on WHO Western Pacific Region and World Federation of Chinese Medicine Societies international standard terminologies on traditional medicine: an analysis of the Diagnostics (part 4). PMID- 20015446 TI - Understanding adolescents' intentions to have the HPV vaccine. AB - From September 2008, older adolescent girls in the UK are eligible to have the HPV vaccine as part of the HPV vaccination 'catch-up' programme. Six months before they were due to be offered the vaccine 16-18 year old girls (N=617) from eight further education/sixth-form colleges reported their intentions to have the HPV vaccine and reasons for this decision. Most intended to have the vaccine (around 70%). Some intentions were influenced by perceptions of risk. Many participants wanted more information and a small minority were scared of needles. Some 'non-intenders' were opposed to vaccinations altogether. The HPV vaccine is likely to be popular but non-intenders' views may be difficult to change. PMID- 20015447 TI - High-performance liquid chromatography assay with fluorescence detection for the evaluation of inhibitors against human recombinant monoacylglycerol lipase. AB - A fluorescent assay for the evaluation of inhibitors of monoacylglycerol lipase (MAGL) is described. 1,3-Dihydroxypropan-2-yl 4-pyren-1-ylbutanoate was designed and synthesized as novel fluorogenic substrate. Activity of human recombinant MAGL was determined in the presence of the surfactant Triton X-100 without further sample cleanup by measuring the amount of 4-pyren-1-ylbutanoic acid released by the enzyme with reversed-phase high-performance liquid chromatography (HPLC) and fluorescence detection. The known covalent binding MAGL inhibitors methyl arachidonyl fluorophosphonate (MAFP), 4-nitrophenyl 4-[bis(1,3-benzodioxol 5-yl)hydroxymethyl]piperidine-1-carboxylate (JZL184), and [4-(5-methoxy-2-oxo 1,3,4-oxadiazol-3-yl)-2-methylphenyl]carbamic acid benzyl ester (CAY10499) were used to validate the test system. Applying an incubation time of 15 min, the IC(50) values obtained for these compounds were 0.16, 3.7, and 1.1 microM, respectively. A prolongation of the incubation to 45 min results in a two- to threefold decrease of the IC(50) values. PMID- 20015448 TI - Vitamin E and rutin synergistically inhibit expression of vascular endothelial growth factor through down-regulation of binding activity of activator protein-1 in human promyelocytic leukemia (HL-60) cells. AB - Reactive oxygen species (ROS) are strong inducers of the angiogenic hormone vascular endothelial growth factor (VEGF). Although, rutin (R) in combination with vitamin E (VE) has been shown to synergistically inhibit oxidative damage, it is unclear whether the combination of R and VE (R+VE) inhibits VEGF secretion in tumor cells. Using a human promyelocytic leukemia (HL-60) cell line, we showed that R in combination with VE synergistically decreased the expressions of VEGF protein and mRNA. We also demonstrated that R+VE significantly decreased the binding capacity of nuclear factor-activator protein-1 (AP-1) to the VEGF gene promoter and decreased the expression of c-Jun protein. Furthermore, we demonstrated that R+VE synergistically reduced insulin receptor substrate-1 (IRS 1) protein expression in HL-60 cells. The decrease of ROS was only partially associated with the decrease of VEGF secreted (r(2)=0.12, P=0.083). Thus, the present results indicate that R in combination with VE attenuates VEGF expression in HL-60 cells and that this effect is mediated by a decreased binding activity of AP-1 through down-regulation of protein expression of insulin-like growth factor 1 receptor (IGF1-R)/IRS-1, while the antioxidant activity of R+VE appears to play a minor role. PMID- 20015449 TI - Toxicity of smoke extracts towards A549 lung cells: role of acrolein and suppression by carbonyl scavengers. AB - The noxious 3-carbon electrophile acrolein forms on combustion of diverse organic matter including synthetic polymers such as polyethylene. While known to play a key role in smoke inhalation injury (SII), the molecular basis for the pulmonary toxicity of high dose acrolein-containing smoke is unclear. As a result, drug interventions in SII are poorly directed against pathogenetic smoke toxicants such as acrolein. The first aim of this study was to confirm a role for acrolein in the acute toxicity of smoke extracts towards A549 lung cells by monitoring adduction of known acrolein targets and the expression of acrolein-inducible genes. A second aim was to evaluate carbonyl scavengers for their abilities to protect cell targets and block smoke extract toxicity. Extracts were prepared by bubbling smoke released by smouldering polyethylene through a buffered saline trap. Acrolein levels in the extracts were estimated via HPLC after derivatisation with 2,4-dinitrophenylhydrazine. Extracts were highly toxic towards A549 cells, eliciting greater ATP depletion than an equivalent concentration of acrolein alone. The toxicity was accompanied by pronounced carbonylation of several cytoskeletal targets, namely vimentin and keratins-7, -8 and -18. Western blotting revealed that polyethylene combustion products also upregulated several acrolein-responsive protein markers, including GADD45beta, NQO1, HMOX, Hsp70, Nur77 and Egr1. Several carbonyl scavengers (bisulfite, d penicillamine, hydralazine and 1-hydrazinoisoquinoline) strongly attenuated smoke extract toxicity, with bisulfite suppressing both the adduction and cross-linking of intermediate filament targets. Bisulfite also suppressed the cytotoxicity of smoke extracts when detected using real-time monitoring of cellular impedance. These findings confirm a key role for acrolein in smoke cytotoxicity and suggest drugs that block acrolein toxicity deserve further investigation as possible interventions against SII. PMID- 20015450 TI - Seasonal reproduction of female round stingrays (Urobatis halleri): steroid hormone profiles and assessing reproductive state. AB - This study characterizes the seasonal reproductive cycle of female round stingrays (Urobatis halleri) in an open coastal site at Seal Beach, CA and a protected estuary at the Seal Beach National Wildlife Refuge (SBNWR). Female round stingrays were sampled from August 2004 to July 2006, and assessed for reproductive parameters (GSI, maximum ova diameter, pregnancy status) and sex steroid (estradiol (E(2)), progesterone (P(4)) and testosterone (T)) concentrations. E(2) and P(4) increased at the time of ovulation (June and July) and remained elevated until parturition (October and November); recently partruded females were observed until November. Mature females were absent from Seal Beach in August and September, the same time period that abundance of mature females peaked in the SBNWR. This aggregation of predominantly mature females in the upper reaches of the SBNWR was seasonal, and was observed from April to September. To better understand the aggregation behavior, sex steroid hormones were assayed in SBNWR females. In July and August, E(2) and P(4) concentrations in females at the SBNWR were 1.5-fold and 2-fold higher, respectively, than concentrations in mature females at Seal Beach, and correlated with elevated water temperature in the estuary. Pregnancy was confirmed in aggregating females by increased levels of E(2) and P(4) and the presence of developing embryos. Our data suggest that coastal estuaries may play a crucial role in round stingray reproduction, perhaps by providing a thermal refuge for pregnant females during gestation. PMID- 20015451 TI - Expression and T3 regulation of thyroid hormone- and sex steroid-related genes during Silurana (Xenopus) tropicalis early development. AB - In amphibians, thyroid hormones (THs) are the primary regulators of metamorphosis; however, their physiological role during embryogenesis remains unclear. First, we established complete developmental profiles for TH receptors (tr alpha and tr beta), deiodinases (dio; types 1, 2, 3), estrogen receptors (er alpha and er beta) and androgen receptor (ar) mRNA levels during embryogenesis and early larval stages in Silurana (Xenopus) tropicalis (from Nieuwkoop and Faber (NF) stage 2 until NF 46). Real-time RT-PCR analyses in whole embryos and larvae revealed that all transcripts except tr alpha were detected throughout development; tr alpha only appears after gastrulation. The first significant increase in the expression of tralpha and tr beta was observed before hatching, between NF 21 and NF 27 (2.5- and 11-fold, respectively). In order to test if these genes could be regulated by THs during early larval development, embryos were exposed to triiodothyronine (T3; 0.5, 5.0, 50 nM) from NF 27 to NF 46. T3 exposure caused a dose-dependent increase relative to control in the expression of tr alpha, tr beta, dio (types 2 and 3), ar, and 5 alpha-reductase type 1 in whole larvae. These results indicate that in S. tropicalis, tr and dio can be induced by T3 as early as NF 46, a response that had only been characterized later during frog metamorphosis. In addition, T3 also affected androgen-related gene expression, supporting our hypothesis that THs are involved in male development in frogs. PMID- 20015452 TI - Basolateral amygdala and morphine-induced taste avoidance in the rat. AB - The present experiment examined the influence of excitotoxic lesions of the basolateral amygdala (BLA) on morphine-induced saccharin avoidance. Neurologically intact subjects rapidly learned to avoid drinking the taste conditioned stimulus (CS), an effect that was sustained throughout the experiment. Although the BLA-lesioned (BLAX) rats showed CS avoidance over the first few trials, the effect was not sustained. That is, by the end of the experiment, the BLAX rats were drinking the same amount of saccharin after seven saccharin-morphine trials as they did on the first trial (i.e., prior to the morphine injections). Potential interpretations of the results are discussed including a disruption of the mechanism that governs drug-induced taste avoidance in normal subjects and the more rapid development of tolerance in BLAX rats. PMID- 20015453 TI - Membrane localization, Caveolin-3 association and rapid actions of vitamin D receptor in cardiac myocytes. AB - The active form of vitamin D, 1alpha, 25-dihydroxyvitamin D(3) (1,25(OH)(2)D(3)), mediates both genomic and rapid non-genomic actions in heart cells. We have previously shown that the vitamin D receptor (VDR) is located in the t-tubular structure of cardiomyocytes. Here we show that VDR specifically interacts with Caveolin-3 in the t-tubules and sarcolemma of adult rat cardiac myocytes. Co immunoprecipitation studies using VDR antibodies revealed that Caveolin-3 specifically co-precipitates with the VDR and similarly the VDR is co precipitated with Caveolin-3 antibody. Confocal immuno-fluorescence microscopy analysis also showed co-localization of VDR and Caveolin-3 in t-tubules and sarcolemma. The non-genomic effects of the functional VDR were studied in electrically stimulated myocytes isolated from adult rat hearts. Sarcomere shortening and re-lengthening were measured in 1,25(OH)(2)D(3) treated cardiac myocytes. A 1nM treatment decreased peak shortening within minutes, suggesting a rapid effect through the membrane-bound VDR. This novel finding of the interaction between VDR and Caveolin-3 is fundamentally important in understanding 1,25(OH)(2)D(3) signal transduction in heart cells and provides further evidence that VDR plays a role in regulation of heart structure and function. PMID- 20015454 TI - Porcine Flt3 ligand and its receptor: generation of dendritic cells and identification of a new marker for porcine dendritic cells. AB - Based on the known importance of Flt3 ligand (Flt3L) for the development of mouse dendritic cells (DCs), the present study compared the phenotype and function of DC derived from porcine bone marrow haematopoietic cells using either granulocyte macrophage colony-stimulating factor or Flt3L (GMCSF-DC and Flt3L-DC, respectively). To this end, porcine Flt3L was cloned resulting in the identification of three isoforms of Flt3L. Compared to GMCSF-DC which were uniformly CD14(+), Flt3L-DC had a more diverse phenotype comprised of CD172a( )CD11a(-) progenitor cells, CD172a(+)CD14(-)CD163(-) DC and CD172a(+)CD14(+)CD163(+) DC. In addition, only the Flt3L-DC contained interferon producing plasmacytoid DC, although their frequency was low. Only the CD14(-) Flt3L-DC responded to TLR2, -3, -4, -7 and -9 agonists by upregulating CD80/86. This population of DC was also more potent in T-cell stimulation assays when compared to the CD14(+) counterpart. Interestingly, Flt3 was not only highly expressed on DC precursors, but also found on Flt3L-DC but not on GMCSF-DC or monocyte-derived DC. Furthermore, also DC circulating in the blood but not monocytes or other leukocytes expressed this receptor. Taken together, our study demonstrates that Flt3L-DCs are more suitable to study the interaction of pathogens with DC. Moreover, we show that also in the pig Flt3 remains expressed in a restricted manner on DC originating from a bone marrow DC precursors, typically representing steady-state DC in lymphoid tissue and blood. PMID- 20015455 TI - Self-reflection and the brain: a theoretical review and meta-analysis of neuroimaging studies with implications for schizophrenia. AB - Several studies have investigated the neural correlates of self-reflection. In the paradigm most commonly used to address this concept, a subject is presented with trait adjectives or sentences and asked whether they describe him or her. Functional neuroimaging research has revealed a set of regions known as Cortical Midline Structures (CMS) appearing to be critically involved in self-reflection processes. Furthermore, it has been shown that patients suffering damage to the CMS, have difficulties in properly evaluating the problems they encounter and often overestimate their capacities and performance. Building on previous work, a meta-analysis of published fMRI and PET studies on self-reflection was conducted. The results showed that two areas within the medial prefrontal cortex (MPFC) are important in reflective processing, namely the ventral (v) and dorsal (d) MPFC. In this paper a model is proposed in which the vMPFC is responsible for tagging information relevant for 'self', whereas the dMPFC is responsible for evaluation and decision-making processes in self- and other-referential processing. Finally, implications of the model for schizophrenia and lack of insight are noted. PMID- 20015456 TI - Opioid modulation of song in male zebra finches (Taenopygia guttata). AB - Endogenous opioids are known to modulate motivated behaviors. Male zebra finches produce a highly motivated behavior (directed song) to court females and also sing in isolation (undirected song). We found that adult male zebra finches sang significantly fewer directed and undirected songs after administration of low doses (2.5 mg/kg body weight) of the general opioid antagonist naloxone, even though the order of syllables in songs was not altered. Surprisingly, high doses of naloxone (10 mg/kg body weight) dramatically decreased the production of undirected songs but had no significant effects on directed songs. There were no changes in the number of calls during directed or undirected song, movement, stereotyped behaviors including pecking and preening, feeding or drinking behaviors after naloxone administration. We also found that treating zebra finches with naloxone led to a decrease in tonality (goodness of pitch), frequency and amplitude modulation and an increase in the length of intersyllable intervals. Our results suggest that the opioid system can differentially modulate directed and undirected song as well as the acoustic characteristics of birdsong, perhaps by acting on different components of the song control system. PMID- 20015458 TI - Flux duality in nonlinear GMA systems: implications for metabolic engineering. AB - Pathway models in biotechnology are customarily designed with metabolite concentrations as the primary, dependent variables, whereas fluxes are derived quantities that are secondarily computed from the primary variables. In other fields of mathematics, such as graph theory and linear systems analysis, it has proven useful to complement primal network model designs in terms of vertices (pools) and edges (processes) with dual designs, where the roles of the primary and secondary quantities are interchanged. The conversion from primal to dual systems is fairly easy in linear systems, but it is unclear to what degree it is possible in nonlinear systems. In this article, we present a method to transform nonlinear primal models of pathways or other biotechnological processes that conform to the Generalized Mass Action (GMA) structure within the formalism of Biochemical Systems Theory (BST) into dual models that focus on fluxes, rather than pools. Interestingly, this transformation is relatively straightforward, once some notational issues are streamlined, and the resulting dual system is again in the format of a GMA system. The transformation is illustrated with the example of glycolysis in Saccharomyces cerevisiae, a well known organism in the food industry. The results suggest how rewriting a model in terms of its fluxes helps bridge the gap between flux balance and dynamic models and also offers a view of the investigated system that is complementary to that of the original model. This dual view is important, because fluxes are sometimes more relevant for the behavior of a biotechnological system than (metabolite) pools. The dual system furthermore offers a systematic approach toward understanding the dynamical constraints under which the system operates. PMID- 20015457 TI - Early postnatal parathion exposure in rats causes sex-selective cognitive impairment and neurotransmitter defects which emerge in aging. AB - Developmental exposure of rats to the organophosphate (OP) pesticides leads to altered neurobehavioral function in juvenile and young adult stages. The current study was conducted to determine whether effects of neonatal parathion exposure on cognitive performance persist in older adult and aged rats, and the relationship of behavioral changes to underlying cholinergic and serotonergic mechanisms. We administered parathion to rat pups on postnatal days 1-4, at doses spanning the threshold for the initial signs of systemic toxicity and for barely detectable cholinesterase inhibition (0.1 or 0.2 mg/kg/day). Beginning at 14 months of age and continuing until 19 months, the rats were trained in the 16-arm radial maze. Controls showed the normal sex difference in this spatial learning and memory task, with the males committing significantly fewer working memory errors than females. Neonatal parathion exposure eliminated the sex difference primarily by causing impairment in males. In association with the effects on cognitive performance, neonatal parathion exposure elicited widespread abnormalities in indices of serotonergic (5HT) and cholinergic synaptic function, characterized by upregulation of 5HT(2) receptors and the 5HT transporter, deficits in choline acetyltransferase activity and nicotinic cholinergic receptors, and increases in hemicholinium-3 binding to the presynaptic choline transporter. Within-animal correlations between behavior and neurochemistry indicated a specific correlation between working memory performance and hippocampal hemicholinium-3 binding; parathion exposure eliminated this relationship. Like the behavioral effects, males showed greater effects of parathion on neurochemical parameters. This study demonstrates the sex-selective, long-term behavioral alterations caused by otherwise nontoxic neonatal exposure to parathion, with effects increasingly expressed with aging. PMID- 20015459 TI - The sea anemone Bunodosoma caissarum toxin BcIII modulates the sodium current kinetics of rat dorsal root ganglia neurons and is displaced in a voltage dependent manner. AB - Sea anemone toxins bind to site 3 of the sodium channels, which is partially formed by the extracellular linker connecting S3 and S4 segments of domain IV, slowing down the inactivation process. In this work we have characterized the actions of BcIII, a sea anemone polypeptide toxin isolated from Bunodosoma caissarum, on neuronal sodium currents using the patch clamp technique. Neurons of the dorsal root ganglia of Wistar rats (P5-9) in primary culture were used for this study (n=65). The main effects of BcIII were a concentration-dependent increase in the sodium current inactivation time course (IC(50)=2.8 microM) as well as an increase in the current peak amplitude. BcIII did not modify the voltage at which 50% of the channels are activated or inactivated, nor the reversal potential of sodium current. BcIII shows a voltage-dependent action. A progressive acceleration of sodium current fast inactivation with longer conditioning pulses was observed, which was steeper as more depolarizing were the prepulses. The same was observed for other two anemone toxins (CgNa, from Condylactis gigantea and ATX-II, from Anemonia viridis). These results suggest that the binding affinity of sea anemone toxins may be reduced in a voltage dependent manner, as has been described for alpha-scorpion toxins. PMID- 20015460 TI - Antimicrobial peptides from the skin secretions of the South-East Asian frog Hylarana erythraea (Ranidae). AB - Peptidomic analysis of norepinephrine-stimulated skin secretions of the South East Asian frog Hylarana erythraea (formerly Rana erythraea partim) has led to the identification of multiple peptides with antimicrobial activity. Structural characterization of the peptides demonstrated that they belong to the brevinin-1 (3), brevinin-2 (2), esculentin-2 (4), and temporin (1) families. The values in parentheses indicate the number of paralogs. In addition, a peptide (GVIKSVLKGVAKTVALG ML.NH(2)) was isolated that shows some structural similarity to the brevinin-2-related peptides (B2RP) previously isolated from North American frogs of the genus Lithobates. A synthetic replicate of the species B2RP showed broad-spectrum growth inhibitory activity against reference strains of Escherichia coli (MIC=12.5 microM), Staphylococcus aureus (MIC=12.5 microM) and Candida albicans (MIC=50 microM) and was active against multidrug-resistant clinical isolates of Acetinobacter baumannii (MIC in the range 6-12.5 microM). The hemolytic activity of the peptide was relatively low (LC(50)=280 microM). Phylogenetic analysis based upon the amino acid sequences of 47 brevinin-2 peptides from 17 Asian species belonging to the family Ranidae provides support for the placement of H. erythraea in the genus Hylarana. PMID- 20015461 TI - Obsessive-compulsive disorder with poor insight: a three-year prospective study. AB - Available evidence about the relationship between poor insight and other clinical characteristics in patients with obsessive-compulsive disorder (OCD) is inconclusive and conflicting. There is also a paucity of data on the long-term course and treatment outcome of OCD patients with poor insight. The present study reports the findings of a relatively large sample (n=106) of outpatients fulfilling DSM-IV criteria for OCD, treated with serotonin reuptake inhibitors (SRIs) and prospectively followed up for 3 years. Baseline information was collected on demographic and clinical characteristics, using standardized instruments. Insight was assessed by means of the Brown Assessment of Beliefs Scale (BABS). Eighty-three patients were followed prospectively and evaluated systematically by validated measures of psychopathology. Compared to their good insight counterparts, the OCD patients with poor insight (22%) showed a greater severity of obsessive-compulsive and depressive symptomatology; an earlier age at onset; a higher rate of schizophrenia spectrum disorder in their first-degree relatives; a higher comorbidity with schizotypal personality disorder. During the follow-up period, poor insight OCD patients were less likely to achieve at least a partial remission of obsessive-compulsive symptoms; required a significantly greater number of therapeutic trials; received more frequently augmentation with antipsychotics. The results suggest that the specifier "poor insight" helps to identify a subgroup of patients at the more severe end of OCD spectrum, characterized by a more complex clinical presentation, a diminished response to standard pharmacological interventions, and a poorer prognosis. Further research is needed to identify alternative strategies for the management of these patients. PMID- 20015462 TI - Association between glycogen synthase kinase-3beta gene polymorphisms and major depression and suicidal behavior in a Korean population. AB - BACKGROUND: Glycogen synthase kinase (GSK)-3beta plays a key role in the phosphorylation and regulation of metabolic enzymes and many transcription factors. Several lines of evidence implicate GSK-3beta in the pathophysiology of mood disorders and susceptibility to suicidal behavior. In this study, we aimed to investigate the GSK-3beta gene's association with major depressive disorder (MDD) and suicidal behavior. METHODS: One hundred seventy suicidal depressed patients and 147 non-suicidal depressed patients who met DSM-IV criteria for MDD were recruited. One hundred sixty-four healthy volunteers recruited by local advertisement served as controls. Patients and normal controls were genotyped for GSK-3beta -1727A/T and -50C/T. Haplotype trend regression (HTR) analysis was used for the evaluation of haplotype association. RESULTS: The genotype distributions of -1727A/T and -50C/T were in agreement with Hardy-Weinberg equilibrium. The results showed that the alleles, genotypes, and haplotypes of the two SNPs do not differ between suicidal MDD subjects, non-suicidal MDD subjects, and normal controls. There was no difference in the haplotype frequency combination between the three groups. CONCLUSION: Our study suggests that two promoter polymorphisms of the GSK-3beta gene may not be related to the pathogenesis of MDD and the risk of suicidal behavior in Korean depressive patients. Further studies with larger sample sizes and different populations are needed. PMID- 20015463 TI - Subchronic mycotoxicoses in Wistar rats: assessment of the in vivo and in vitro genotoxicity induced by fumonisins and aflatoxin B(1), and oxidative stress biomarkers status. AB - Some evidence suggests that fumonisin B(1) (FB(1)), a worldwide toxic contaminant of grains produced by Fusarium verticillioides, exhibits an oxidative stress mediated genotoxicity. We studied the DNA damage (by the alkaline comet and the micronucleus tests) and biomarkers of cellular oxidative stress (malondialdehyde, MDA; catalase, CAT; and superoxide dismutase, SOD) in spleen mononuclear cells of male Wistar rats subchronically (90 days) fed on a control experimental diet (CED) or poisoned with experimental diets contaminated with a culture material containing 100 ppm of FB(1) (FED), with 40 ppb of aflatoxin B(1) (a common toxic co-contaminant in cereals, AFB(1)ED), and with a mixture of both toxins (MED). The DNA damage was found in 13.7%, 81.7%, 98.0% and 99.3% (comet assay) and in 2.8%, 7.0%, 10.8% and 8.8% (micronucleus technique) in groups CED, FED, AFB(1)ED and MED, respectively. The MDA levels as well as the CAT and SOD activities were increased in all the poisoned animals. A similar behavior was observed in cells exposed in vitro to the toxins. These data support the hypothesis of an oxidative stress mediated genotoxicity induced by FB(1). Furthermore, the extent of DNA damage assessed by the comet assay suggests a possible protective effect of the fumonisins-AFB(1) mixtures in vitro against the genotoxicity induced individually by the toxins. PMID- 20015464 TI - T3 acutely increases GH mRNA translation rate and GH secretion in hypothyroid rats. AB - Cytoskeleton controls the stability of transcripts, by mechanisms that involve mRNAs and eEF1A attachment to it. Besides, it plays a key role in protein synthesis and secretion, which seems to be impaired in somatotrophs of hypothyroid rats, whose cytoskeleton is disarranged. This study investigated the: eEF1A and GH mRNA binding to cytoskeleton plus GH mRNA translation rate and GH secretion, in sham-operated and thyroidectomized rats treated with T3 or saline, and killed 30min thereafter. Thyroidectomy reduced: (a) pituitary F-actin content, and eEF1A plus GH mRNA binding to it; (b) GH mRNA recruitment to polysome; and (c) liver IGF-I mRNA expression, indicating that GH mRNA stability and translation rate, as well as GH secretion were impaired. T3 acutely reversed all these changes, which points toward a nongenomic action of T3 on cytoskeleton rearrangement, which might contribute to the increase on GH mRNA translation rate and GH secretion. PMID- 20015465 TI - Expression of IFNalpha-inducible genes and modulation of HLA-DR and thyroid stimulating hormone receptors in Graves' disease. AB - Interferon alpha (IFNalpha) plays an important role in the pathogenesis of different autoimmune diseases. IFNalpha is widely used for the treatment of chronic viral infections, particularly chronic hepatitis C virus infection; however, several case reports have emerged describing autoimmune conditions, such as Graves' disease (GD), that have developed in the patients receiving IFNalpha. The mechanism by which IFNalpha is involved in GD remains poorly understood. We investigated the expression of IFNalpha and IFNalpha-inducible genes (IFIGs) in GD and found that IFIGs were overexpressed in 60% of 54 clinical diagnostic GD patients. These elevated IFIGs correlated with serological levels of autoantibody to thyroid stimulating hormone receptor (TSHR). Recombinant human IFNalpha stimulated primary cultured thyrocytes resulted in not only high level expression of IFIGs, but also, more importantly, expression of MHC-II antigens (HLA-DR3 and HLA-DR5) and TSHR in GD subjects. Furthermore, thyroid gland tissues from GD patients over express HLA-DR, TSHR and IFNalpha receptors at both messenger RNA and protein levels. Taken together, these data indicated that in GD patients, IFNalpha can function on thyroid tissue to induce a number of genes, particularly MHC class II molecules which may enhance autoantigen presentation of TSHR on thyrocytes. PMID- 20015466 TI - Selective D1 agonism but not D2 antagonism is reflected in cAMP and cGMP levels in rat CSF. AB - Cyclic adenosine 3'5'-monophosphate (cAMP) and cyclic guanosine 3'5' monophosphate (cGMP) serve as second messengers in several cellular pathways within the central nervous system. In various neurological and psychiatric disorders with known deficits in neurotransmission function, CSF levels of cAMP and/or cGMP in patients were studied. Very little information is currently available on cAMP and cGMP levels in CSF of animals. Moreover, this is the first study on the effects of pharmacological treatment on cAMP and cGMP levels in rat CSF. Effect of systemic treatment with a D1 receptor agonist SKF82958 and a D2 receptor antagonist haloperidol on cAMP and cGMP levels, as well as baseline cAMP and cGMP levels in CSF of rats was determined. A significantly increased cAMP and cGMP level in cisternal CSF of rats systemically treated with the D1 receptor agonist SKF82958 was observed, while when treated with the D2 antagonist haloperidol, no effect on cAMP and only a slight decrease of cGMP was observed after treatment with the highest dose. Determining cAMP and/or cGMP in CSF of experimental animals can serve as a useful tool to study neural processes affected by disease and treatment. PMID- 20015467 TI - Context affects the numerical semantic congruity effect in rhesus monkeys (Macaca mulatta). AB - Do monkeys anchor their numerical judgments based on the context in which their choices are presented? We addressed this question by varying the numerical range across sessions while macaque monkeys made ordinal judgments. Monkeys were trained to make a conditional discrimination whereby they were reinforced for ordering arrays of dots in ascending or descending numerical order, dependent on a color cue. Monkeys were tested using two ranges of numerosities that converged on a single pair. Similar to the findings of Cantlon and Brannon (2005), we found a semantic congruity effect whereby decision time was systematically influenced by the congruity between the cue (ascending or descending) and the relative Numerical Magnitude of the stimuli within each range. Furthermore, monkeys showed a context effect, such that decision time for a given pair was dependent on whether it was a relatively small or large set of values compared to the other values presented in that session. This finding suggests that, similar to humans, the semantic congruity effect observed in monkeys is anchored by the context. Thus our data provide further evidence for the existence of a shared numerical comparison process in monkeys and humans. PMID- 20015468 TI - Learning and anticipatory behaviour in a "sit-and-wait" predator: the Atlantic halibut. AB - We studied the learning capacities and anticipatory behaviour in a "sit-and-wait" predatory fish, the Atlantic halibut, Hippoglossus hippoglossus. In Experiment 1 two groups of halibut received series of light flashes (conditioned stimulus, CS) that started before delivery of food (unconditioned stimulus, US) and persisted until after food delivery, i.e. delay conditioning. Control groups received unpaired CS and US presentations. The anticipatory behaviour of delay conditioned halibut consisted mainly of take-offs towards the surface shortly after onset of the CS. In Experiment 2 six groups of halibut were trained in three trace conditioning procedures: Two groups with 20s, two groups with 60s and two groups with 120s trace interval. Learning was evident in the 20 and 60s trace groups and in one of the 120s trace groups. In contrast to delay conditioning the anticipatory behaviour of trace conditioned halibut was characterized by subtle movements near the tank floor with orientation towards the CS. The cautious responses of halibut after trace conditioning differed markedly from what is observed in other fish species and are suggested to reflect a "sit-and-wait" foraging strategy that requires the predator to remain undetected until the prey is within lunging range. PMID- 20015469 TI - Ototoxic drugs: difference in sensitivity between mice and guinea pigs. AB - The development of experimental animal models has played an invaluable role in understanding the mechanisms of neurosensory deafness and in devising effective treatments. The purpose of this study was to develop an adult mouse model of ototoxic drug-induced hearing loss and to compare the ototoxicity in the adult mouse to that in the well-described guinea pig model. Mice are a powerful model organism, especially due to the large availability of antibodies, probes and genetic mutants. In this study, mice (n=114) and guinea pigs (n=35) underwent systemic treatment with either kanamycin or cisplatin. Auditory brainstem responses showed a significant threshold shift in guinea pigs 2 weeks after the beginning of the ototoxic treatment, while there was no significant hearing impairment recorded in mice. Hair cells and neuronal loss were correlated with hearing function in both guinea pigs and mice. These results indicate that the mouse is not a good model for ototoxicity, which should be taken into consideration in all further investigations concerning ototoxicity-induced hearing loss. PMID- 20015470 TI - Blending of agglomerates into powders 1: Quantification of abrasion rate. AB - A very common situation in the pharmaceutical arena is that a small amount of cohesive drug substance needs to be distributed in a large bulk of free-flowing filler such as lactose. The key topic of attention is that aggregates of a cohesive drug substance need to be sufficiently broken up in an acceptable time frame. This implies that there is need for a better mechanistic understanding of the blending process and the reduction in size of the aggregates. The purpose of this study is to obtain more insight in the mechanisms that lead to the break up of assemblies of powder particles in a moving powder bed. The break up of aggregates was studied by application of so-called brittle Calibrated Test Particles (bCTPs). These are well-defined aggregates with brittle fracture properties. The dominant mechanism of the break up of these aggregates is abrasion by multiple impacts. There is evidence of a relationship between strength (expressed as porosity) of the bCTPs and rate of abrasion. This is often a slow process and the rate is determined not only by the (mechanical) properties of the agglomerates and process conditions, but also by the particle size distribution of the bulk filler. PMID- 20015471 TI - Fuzheng Huayu recipe and vitamin E reverse renal interstitial fibrosis through counteracting TGF-beta1-induced epithelial-to-mesenchymal transition. AB - AIM: To investigate the mechanism of action of Fuzheng Huayu recipe (FZHY) and vitamin E (Vit E) against renal interstitial fibrosis related to transforming growth factor-beta1 (TGF-beta1) mediated tubular epithelial-to-mesenchymal transition. MATERIALS AND METHODS: Renal interstitial fibrosis was induced by administration of HgCl(2) at a dose of 8 mg/kg body weight once a day for 9 weeks. Rats were randomly divided into four groups: normal, model, FZHY, and Vit E group. Rats in the latter two groups were treated with the FZHY recipe and Vit E respectively. HK-2 cells were treated with TGF-beta1 for 24h, followed by incubation with either SB-431542 (a potent and specific inhibitor of TbetaR-I kinase) or FZHY drug-containing serum for another 24h. Hyp content in rat kidney tissue was assayed with Jamall's method and collagen deposition in kidney was visualized using Masson stain. Protein expression of TGF-beta1, TbetaR-I, Smad2, p-Smad2, Smad3, and p-Smad3 was analyzed by Western blotting. Protein expression and the location of Smad3 in kidney was assayed by immunohistochemistry, E cadherin, cytokeratin 18 (CK-18), alpha-SMA and TGF-beta1 by immunofluorescent stain. RESULTS: FZHY and Vit E inhibited renal collagen deposition and reduced Hyp content significantly. They upregulated E-cadherin protein expression and down-regulated the protein expression of alpha-SMA, TGF-beta1, p-Smad2, p-Smad3, and TbetaR-I. Lastly, they inhibited the nuclear translocation of Smad3 in fibrotic kidney tissue. FZHY drug-containing serum significantly upregulated the expression of CK-18 and down-regulated the expression of alpha-SMA, TbetaR-I, p Smad2/3 in TGF-beta1 stimulated HK-2 cells. CONCLUSION: The mechanism of action of FZHY and Vit E against renal interstitial fibrosis is related to the reversal of tubular EMT induced by TGF-beta1. PMID- 20015472 TI - Metabolic intervention of aflatoxin B1 toxicity by curcumin. AB - ETHNO PHARMACOLOGICAL RELEVANCE: Curcumin, bioactive principle of turmeric (Curcuma longa Linn) is an important constituent of Indian traditional medicine. Turmeric has been known to possess several therapeutic properties. AIM OF THE STUDY: The modulatory effect of dietary curcumin (0.05%, w/w) on drug metabolizing and general marker enzymes of liver and formation of AFB(1)-adducts (DNA and protein) due to dietary AFB(1) exposure for a period of 6 weeks in a rodent model, have been evaluated. MATERIALS AND METHODS: Drug metabolizing enzymes CYP1A1, GSHT, UGT1A and general marker enzymes (LDH, ALT, AST, ALP and gamma-GT) of liver were estimated by standardized methods. Aflatoxin adducts (DNA and protein) were quantitated by indirect competitive ELISA. RESULTS: Dietary curcumin enhanced GSHT (p<0.001) and UGT1A1 (p<0.05) activity and significantly reduced the activity of CYP1A1 (p<0.001), in rats exposed to aflatoxin B(1). Supplementation of curcumin in the diet normalized the altered activities of LDH and ALT. At molecular level, curcumin significantly reduced AFB(1)-N(7)-guanine adduct (p<0.001) excretion in the urine, DNA adduct (p<0.05) in the liver and albumin adduct (p<0.001) in the serum. CONCLUSION: The experimental results substantiates that curcumin intervention ameliorates the AFB(1) induced toxicity. PMID- 20015473 TI - Shu-Gan-Liang-Xue Decoction, a Chinese herbal formula, down-regulates the expression of steroid sulfatase genes in human breast carcinoma MCF-7 cells. AB - ETHNOPHARMACOLOGICAL RELEVANCE: Shu-Gan-Liang-Xue Decoction (SGLXD), a traditional Chinese herbal formula, has been used to ameliorate hot flushes symptom in breast cancer patients for decades. AIM OF THE STUDY: Steroid sulfatase (STS) has a crucial role in regulating the estrogen biosynthesis within breast tumors. We aimed to investigate whether SGLXD could modulate STS expression in human breast carcinoma MCF-7 cells. MATERIALS AND METHODS: By semi quantitative/quantitative reverse transcription-PCR, we investigated the transcript level of STS in MCF-7 cells treated with various concentrations of SGLXD. By using transient cotransfection with estrogen dependent plasmid pERE-TK Luc and internal control plasmid pRL-TK in MCF-7 cells and dual luciferase reporter (DLR) based bioluminescent measurements, we evaluated the enzymatic activity of STS after SGLXD treatment. RESULTS: By RT-PCR and real time PCR, the mRNA level of STS was decreased by SGLXD treatment, in the dose-dependent manner, compared to negative control (p<0.01). By DLR assay, different concentrations of SGLXD significantly inhibited the enzymatic activity of STS in MCF-7 cells dose dependently (p<0.05). CONCLUSIONS: SGLXD could act as a selective estrogen enzyme modulator by down-regulating the STS expression in MCF-7 cells. PMID- 20015474 TI - Long-term effects of high lipid and high energy diet on serum lipid, brain fatty acid composition, and memory and learning ability in mice. AB - OBJECTIVE: It is well known that high lipid and high energy diet is harmful to health. But the different effects of high lipid diet composed of either saturated fatty acids or unsaturated fatty acids have not been distinguished. METHOD: Eighteen pregnant C57BL/6j (22-25g) mice were randomly divided into three groups of six each and fed with chow or high lipid diet composed of either flaxseed oil (chow diet 84%, cholesterol 0.2%, flaxseed oil 15.8%) or lard fat (chow diet 84%, cholesterol 0.2%, lard fat 15.8%). After weaning, the offspring were fed the same diet as their mothers were fed during the experiment, and their spatial memory and learning ability were evaluated by Morris water maze when they were 8 weeks old. Next, the blood and tissues were sampled when they were 9 weeks old. Serum lipids were determined using kits, and brain fatty acids were measured using a gas chromatograph. RESULTS: Compared to chow diet (control), high flaxseed oil diet (HFO) increased high density lipoprotein cholesterol level (HDL-C) in the mothers but not in offspring; high lard fat diet (HLF) increased serum total cholesterol level (TC) and low density lipoprotein cholesterol level (LDL-C) both in mothers and offspring. Brain fatty acids profile was altered by HLF compared with chow diet. Polyunsaturated fatty acids and long-chain polyunsaturated fatty acids content were significantly lower in the HLF group than in the control group, but saturated fatty acids content were significantly higher in HLF group than those in control group. The changed fatty acids composition affected the spatial memory and learning ability of adult offspring. CONCLUSIONS: A long-term high lard diet increased offspring serum TC and LDL-C levels and affected the brain's fatty acid composition, and memory and learning ability. The polyunsaturated fatty acid content of the brain may be correlated with serum cholesterol levels. PMID- 20015475 TI - Prostaglandin E2 activates cAMP response element-binding protein in glioma cells via a signaling pathway involving PKA-dependent inhibition of ERK. AB - Prostaglandin E(2) (PGE(2)) plays a critical role in influencing the biological behavior of tumor cells. We previously demonstrated that PGE(2) stimulates human glioma cell growth via activation of protein kinase A (PKA) type II. This study was undertaken to further elucidate the intracellular pathways activated by PGE(2) downstream to PKA. Stimulation of U87-MG glioma cells with PGE(2) increased phosphorylation of the cyclic-AMP response element (CRE) binding protein CREB at Ser-133 and CREB-driven transcription in a dose- and time dependent manner. Expression of dominant CREB constructs that interfere with CREB phosphorylation at Ser-133 or with its binding to the CRE site markedly decreased PGE(2)-induced CREB activation. Inhibition of PKA by H-89 or expression of a dominant negative PKA construct attenuated PGE(2)-induced CREB activation. Moreover, inhibition of PKA type II decreased PGE(2)-induced CREB-dependent transcription by 45% compared to vehicle-treated cells. To investigate the involvement of additional signaling pathways, U87-MG cells were pretreated with wortmannin or LY294002 to inhibit the PI3-kinase/AKT pathway. Both inhibitors had no effect on PGE(2)-induced CREB phosphorylation and transcriptional activity, suggesting that PGE(2) activates CREB in a PI3-kinase/AKT independent manner. Challenge of U87-MG cells with PGE(2), at concentrations that induced maximal CREB activation, or with forskolin inhibited extracellular signal-regulated kinase (ERK) phosphorylation. Pretreatment of U87-MG cells with the ERK inhibitor PD98059, accentuated ERK inhibition and increased CREB phosphorylation at Ser-133 and CREB-driven transcription stimulated by PGE(2), suggesting that inhibition of ERK contributes to PGE(2)-induced CREB activation. Inhibition of ERK by PGE(2) or by forskolin was rescued by treatment of cells with H-89 or by the dominant negative PKA construct. Moreover, PGE(2) or forskolin inhibited phosphorylation of Raf-1 phosphorylation at Ser-338. Challenge of U87-MG cells with 11-deoxy PGE(1) increased CREB-driven transcription and stimulated cell growth, while other PGE(2) analogues had no effect. Together our results reveal a novel signaling pathway whereby PGE(2) signals through PKA to inhibit ERK and increase CREB transcriptional activity. PMID- 20015476 TI - DNA damage in earthworms from highly contaminated soils: assessing resistance to arsenic toxicity by use of the Comet assay. AB - Earthworms native to the former mine site of Devon Great Consols (DGC), UK reside in soils highly contaminated with arsenic (As). These earthworms are considered to have developed a resistance to As toxicity. The mechanisms underlying this resistance however, remain unclear. In the present study, non-resistant, commercially sourced Lumbricus terrestris were exposed to a typical DGC soil in laboratory mesocosms. The earthworms bio-accumulated As from the soil and incurred DNA-damage levels significantly above those observed in the control mesocosm (assessed using the Comet assay). A dose response was observed between DNA damage (% tail DNA) and As concentration in soil (control, 98, 183, 236, 324 and 436mgkg(-1)). As-resistant earthworms (Lumbricus rubellus, Dendrodrilus rubidus and L. terrestris) collected from contaminated soils at DGC (203 to 9025mgkg(-1) As) had also bio-accumulated high levels of As from their host soils, yet demonstrated low levels of DNA damage compared with earthworms from uncontaminated sites. The results demonstrate that the As-contaminated soils at DGC are genotoxic to non-native earthworms and much less so to earthworms native to DGC, thus providing further evidence of an acquired resistance to As toxicity in the native earthworms. PMID- 20015477 TI - The effect of oxidative stress on nucleotide-excision repair in colon tissue of newborn piglets. AB - Nucleotide-excision repair (NER) is important for the maintenance of genomic integrity and to prevent the onset of carcinogenesis. Oxidative stress was previously found to inhibit NER in vitro, and dietary antioxidants could thus protect DNA not only by reducing levels of oxidative DNA damage, but also by protecting NER against oxidative stress-induced inhibition. To obtain further insight in the relation between oxidative stress and NER activity in vivo, oxidative stress was induced in newborn piglets by means of intra-muscular injection of iron (200mg) at day 3 after birth. Indeed, injection of iron significantly increased several markers of oxidative stress, such as 8-oxo-7,8 dihydro-2'-deoxyguanosine (8-oxodG) levels in colon DNA and urinary excretion of 8-oxo-7,8-dihydroguanine (8-oxoGua). In parallel, the influence of maternal supplementation with an antioxidant-enriched diet was investigated in their offspring. Supplementation resulted in reduced iron concentrations in the colon (P=0.004) at day 7 and a 40% reduction of 8-oxodG in colon DNA (P=0.044) at day 14 after birth. NER capacity in animals that did not receive antioxidants was significantly reduced to 32% at day 7 compared with the initial NER capacity on day 1 after birth. This reduction in NER capacity was less pronounced in antioxidant-supplemented piglets (69%). Overall, these data indicate that NER can be reduced by oxidative stress in vivo, which can be compensated for by antioxidant supplementation. PMID- 20015478 TI - Evaluation of probabilistic and logical inference for a SNP annotation system. AB - Genome wide association studies (GWAS) are an important approach to understanding the genetic mechanisms behind human diseases. Single nucleotide polymorphisms (SNPs) are the predominant markers used in genome wide association studies, and the ability to predict which SNPs are likely to be functional is important for both a priori and a posteriori analyses of GWA studies. This article describes the design, implementation and evaluation of a family of systems for the purpose of identifying SNPs that may cause a change in phenotypic outcomes. The methods described in this article characterize the feasibility of combinations of logical and probabilistic inference with federated data integration for both point and regional SNP annotation and analysis. Evaluations of the methods demonstrate the overall strong predictive value of logical, and logical with probabilistic, inference applied to the domain of SNP annotation. PMID- 20015479 TI - Naive predictions of motion and orientation in mirrors: From what we see to what we expect reflections to do. AB - The study aimed to investigate naive beliefs regarding the dynamic and static behavior of reflections. In the first three experiments, participants in the study made predictions about the correspondence between real and reflected movements or about the orientation of the reflection of a static object placed in front of a mirror. In Experiments 1 and 2, paper-and-pencil tasks were used and in Experiment 3 participants were asked to make their predictions while imagining that they were facing a mirror. Results revealed that a percentage of undergraduates (ranging from 25% to 35%) were unable to make correct predictions. We classified the errors into types and found that responses either conform to the belief that reflections do the same or that they do the opposite. This suggests an oversimplification of the geometry of mirror reflections in two directions: participants either generalize what they see when movements are parallel to the mirror or what they see when movements are orthogonal to the mirror. Findings from Experiment 4 confirmed that these two expectations fit in with what people perceive in mirrors. Findings from Experiment 5 confirmed that this is also in agreement with the relationship perceived when looking at similar movements and orientations "outside" mirrors. PMID- 20015480 TI - Reducing control selection errors associated with underground bolting equipment. AB - Selecting the incorrect control during the operation of underground bolting and drilling equipment causes serious injuries. Shape coding and the layout of dual control banks are two aspects of control design which require further examination. The aims of this research were: (i) to determine whether arbitrary shape coding was effective in reducing selection error rates in a virtual analogy of roof-bolting; and (ii) to determine whether any advantages exist for mirror or place layouts for dual control situations in this situation. Two experiments were conducted to address these questions. No benefits of arbitrary shape coding were evident while control location remained constant. When control location was altered, shape coding did provide a significant reduction in selection error rate. No differences between mirror or place arrangements were detected and this question remains open. PMID- 20015482 TI - Gene-environment interactions and response to social intrusion in male and female rhesus macaques. AB - BACKGROUND: Genetic factors interact with environmental stressors to moderate risk for human psychopathology, but sex may also be an important mediating factor. Different strategies for coping with environmental stressors have evolved in males and females, and these differences may underlie the differential prevalence of certain types of psychopathology in the two sexes. In this study, we investigated the possibility of sex-specific gene-environment interactions in a nonhuman primate model of response to social threat. METHODS: Rhesus macaques (77 males and 106 females) were exposed to an unfamiliar conspecific. Using factor analysis, we identified three behavioral factors characterizing the response to social threat. Monkeys were genotyped for the serotonin transporter linked polymorphism (5-HTTLPR), and the effects of genotype, early life stress, and sex on behavioral responses were evaluated. RESULTS: Factor analysis produced five factors: High-Risk Aggression, Impulsivity/Novelty-Seeking, Gregariousness/Boldness, Harm Avoidance, and Redirected Aggression. Overall, males displayed higher levels of High-Risk Aggression and Gregariousness/Boldness than females. Levels of High-Risk Aggression in males carrying the s allele were significantly higher if they were also exposed to early adversity in the form of peer rearing. CONCLUSIONS: Our findings support those from studies in humans suggesting that males are more vulnerable to externalizing or aggression-related disorders. The results highlight the importance of interactions that exist among behavior, genes, and the environment and suggest that sex differences in vulnerability to psychopathology may be grounded in our evolutionary history. PMID- 20015481 TI - Neuroplasticity in the mesolimbic system induced by natural reward and subsequent reward abstinence. AB - BACKGROUND: Natural reward and drugs of abuse converge on the mesolimbic system, where drugs of abuse induce neuronal alterations. Here, we tested plasticity in this system after natural reward and the subsequent impact on drug responses. METHODS: Effects of sexual experience in male rats on behavioral sensitization and conditioned place preference associated with d-amphetamine (AMPH) and Golgi impregnated dendrites and spines of nucleus accumbens (NAc) cells were determined. Moreover, the impact of abstinence from sexual behavior in experienced males on these parameters was tested. RESULTS: First, repeated sexual behavior induced a sensitized locomotor response to AMPH compared with sexually naive control subjects observed 1, 7, and 28 days after last mating session. Second, sexually experienced animals formed a conditioned place preference for lower doses of AMPH than sexually naive males, indicative of enhanced reward value of AMPH. Finally, Golgi-Cox analysis demonstrated increased numbers of dendrites and spines in the NAc core and shell with sexual experience. The latter two alterations were dependent on a period of abstinence of 7-10 days. CONCLUSIONS: Sexual experience induces functional and morphological alterations in the mesolimbic system similar to repeated exposure to psychostimulants. Moreover, abstinence from sexual behavior after repeated mating was essential for increased reward for drugs and dendritic arbors of NAc neurons, suggesting that the loss of sexual reward might also contribute to neuroplasticity of the mesolimbic system. These results suggest that some alterations in the mesolimbic system are common for natural and drug reward and might play a role in general reinforcement. PMID- 20015483 TI - Hippocampal changes associated with early-life adversity and vulnerability to depression. AB - BACKGROUND: Smaller hippocampal volume has been reported in some adult and pediatric studies of unipolar major depressive disorder. It is not clear whether the smaller hippocampal volume precedes or is a consequence of the illness. Early life adversity is associated with both smaller hippocampal volume and increased vulnerability to depressive disorder. Hippocampal changes may mediate the relationship between early-life adversity and depressive illness in a subset of patients. However, there are no reports of longitudinal clinical studies that have examined this issue. METHODS: Thirty adolescents with unipolar major depressive disorder, 22 adolescent volunteers with no personal history of a psychiatric illness including depression but who were at high risk for developing depression by virtue of parental depression (high-risk group), and 35 adolescent volunteers with no personal or family history of a psychiatric disorder (control subjects) underwent volumetric magnetic resonance imaging studies. Information was also gathered on early and recent adverse experiences with standard interviews. The participants were followed for up to 5 years to assess the onset and clinical course of depression. RESULTS: Depressed and high-risk groups had significantly smaller left and right hippocampal volumes than control subjects. Higher levels of early-life adversity were associated with smaller hippocampal volumes. Smaller hippocampal volume partially mediated the effect of early-life adversity on depression during longitudinal follow-up. CONCLUSIONS: Smaller hippocampal volume in adolescents at high risk for depression suggests that it may be a vulnerability marker for the illness. Early-life adversity may interact with genetic vulnerability to induce hippocampal changes, potentially increasing the risk for depressive disorder. PMID- 20015484 TI - Slc25a12 disruption alters myelination and neurofilaments: a model for a hypomyelination syndrome and childhood neurodevelopmental disorders. AB - BACKGROUND: SLC25A12, a susceptibility gene for autism spectrum disorders that is mutated in a neurodevelopmental syndrome, encodes a mitochondrial aspartate glutamate carrier (aspartate-glutamate carrier isoform 1 [AGC1]). AGC1 is an important component of the malate/aspartate shuttle, a crucial system supporting oxidative phosphorylation and adenosine triphosphate production. METHODS: We characterized mice with a disruption of the Slc25a12 gene, followed by confirmatory in vitro studies. RESULTS: Slc25a12-knockout mice, which showed no AGC1 by immunoblotting, were born normally but displayed delayed development and died around 3 weeks after birth. In postnatal day 13 to 14 knockout brains, the brains were smaller with no obvious alteration in gross structure. However, we found a reduction in myelin basic protein (MBP)-positive fibers, consistent with a previous report. Furthermore, the neocortex of knockout mice contained abnormal neurofilamentous accumulations in neurons, suggesting defective axonal transport and/or neurodegeneration. Slice cultures prepared from knockout mice also showed a myelination defect, and reduction of Slc25a12 in rat primary oligodendrocytes led to a cell-autonomous reduction in MBP expression. Myelin deficits in slice cultures from knockout mice could be reversed by administration of pyruvate, indicating that reduction in AGC1 activity leads to reduced production of aspartate/N-acetylaspartate and/or alterations in the dihydronicotinamide adenine dinucleotide/nicotinamide adenine dinucleotide(+) ratio, resulting in myelin defects. CONCLUSIONS: Our data implicate AGC1 activity in myelination and in neuronal structure and indicate that while loss of AGC1 leads to hypomyelination and neuronal changes, subtle alterations in AGC1 expression could affect brain development, contributing to increased autism susceptibility. PMID- 20015485 TI - Anatomical alterations and symptom-related functional activity in obsessive compulsive disorder are correlated in the lateral orbitofrontal cortex. PMID- 20015486 TI - A meta-analysis of cytokines in major depression. AB - BACKGROUND: Major depression occurs in 4.4% to 20% of the general population. Studies suggest that major depression is accompanied by immune dysregulation and activation of the inflammatory response system (IRS). Our objective was to quantitatively summarize the data on concentrations of specific cytokines in patients diagnosed with a major depressive episode and controls. METHODS: We performed a meta-analysis of studies measuring cytokine concentration in patients with major depression, with a database search of the English literature (to August 2009) and a manual search of references. RESULTS: Twenty-four studies involving unstimulated measurements of cytokines in patients meeting DSM criteria for major depression were included in the meta-analysis; 13 for tumor necrosis factor (TNF)-alpha, 9 for interleukin (IL)-1beta, 16 for IL-6, 5 for IL-4, 5 for IL-2, 4 for IL-8, 6 for IL-10, and 4 for interferon (IFN)-gamma. There were significantly higher concentrations of TNF-alpha (p < .00001), weighted mean difference (WMD) (95% confidence interval) 3.97 pg/mL (2.24 to 5.71), in depressed subjects compared with control subjects (438 depressed/350 nondepressed). Also, IL-6 concentrations were significantly higher (p < .00001) in depressed subjects compared with control subjects (492 depressed/400 nondepressed) with an overall WMD of 1.78 pg/mL (1.23 to 2.33). There were no significant differences among depressed and nondepressed subjects for the other cytokines studied. CONCLUSIONS: This meta-analysis reports significantly higher concentrations of the proinflammatory cytokines TNF-alpha and IL-6 in depressed subjects compared with control subjects. While both positive and negative results have been reported in individual studies, this meta-analytic result strengthens evidence that depression is accompanied by activation of the IRS. PMID- 20015487 TI - Cross-modal facilitation in speech prosody. AB - Speech prosody has traditionally been considered solely in terms of its auditory features, yet correlated visual features exist, such as head and eyebrow movements. This study investigated the extent to which visual prosodic features are able to affect the perception of the auditory features. Participants were presented with videos of a speaker pronouncing two words, with visual features of emphasis on one of these words. For each trial, participants saw one video where the two words were identical in both pitch and amplitude, and another video where there was a difference in either pitch or amplitude that was congruent or incongruent with the visual changes. Participants were asked to decide which video contained the sound difference. Thresholds were obtained for the congruent and incongruent videos, and for an auditory-alone condition. It was found that the congruent thresholds were better than the incongruent thresholds for both pitch and amplitude changes. Interestingly, the congruent thresholds for amplitude were better than for the auditory-alone condition, which implies that the visual features improve sensitivity to loudness changes. These results demonstrate that visual stimuli can affect auditory thresholds for changes in pitch and amplitude, and furthermore support the view that visual prosodic features enhance speech processing. PMID- 20015488 TI - Evidence of seasonal variation in longitudinal growth of height in a sample of boys from Stuttgart Carlsschule, 1771-1793, using combined principal component analysis and maximum likelihood principle. AB - Recent progress in modelling individual growth has been achieved by combining the principal component analysis and the maximum likelihood principle. This combination models growth even in incomplete sets of data and in data obtained at irregular intervals. We re-analysed late 18th century longitudinal growth of German boys from the boarding school Carlsschule in Stuttgart. The boys, aged 6 23 years, were measured at irregular 3-12 monthly intervals during the period 1771-1793. At the age of 18 years, mean height was 1652 mm, but height variation was large. The shortest boy reached 1474 mm, the tallest 1826 mm. Measured height closely paralleled modelled height, with mean difference of 4 mm, SD 7 mm. Seasonal height variation was found. Low growth rates occurred in spring and high growth rates in summer and autumn. The present study demonstrates that combining the principal component analysis and the maximum likelihood principle enables growth modelling in historic height data also. PMID- 20015489 TI - Characteristics and outcomes of patients discharged home from the Emergency Department following trauma team activation. AB - OBJECTIVE: Past research on trauma teams has largely focused on the outcomes of severely injured patients. Few studies have looked at patients who have activated the trauma team but are discharged home directly from the Emergency Department. The aim of this study was to examine the characteristics and outcomes of these patients following discharge. METHODS: All adult Emergency Department discharged trauma patients who were contactable by telephone 7-14 days post-discharge and spoke English were eligible for the study. A 10-min questionnaire was conducted covering their perceptions of Emergency Department care, return to activities, discharge and follow-up care, missed injuries and pain management. Data were also collected on their age, sex, injuries and length of stay in the Emergency Department. RESULTS: Over the 169-day study period, 158 trauma patients were discharged from Liverpool hospital, which formed 30.1% of all patients treated by the trauma team. Of these, 106 patients were contactable and 100 completed the follow-up questionnaire. They suffered mainly minor injuries but stayed a median 341 min in the Department. Most patients (87%) reported that their health had impacted on their daily activities and about half of all full-time workers remained off work for 1 week or more. A small number of patients had missed fractures or other serious injuries. Two-third of patients visited a medical practitioner after discharge and 8 required further specialist and/or in-patient care. CONCLUSION: Most trauma patients discharged from the Emergency Department continue to suffer significant morbidity after their departure from hospital and require further medical care. A small number of patients also had significant missed injuries. This suggests that more comprehensive discharge and follow-up care for these patients is warranted. PMID- 20015490 TI - Transabdominal amnioinfusion in preterm premature rupture of membranes. AB - OBJECTIVE: To evaluate the effect of transabdominal amnioinfusion on prolongation of pregnancy, and maternal and neonatal outcomes in preterm premature rupture of membranes (pPROM). METHODS: We conducted a prospective randomized controlled study of women with pPROM during singleton live pregnancy-between 26 and 33+6weeks-whose amniotic fluid index (AFI) was less than the 5th percentile. The study group underwent transabdominal amnioinfusion at admission and then weekly if their AFI fell below the 5th percentile again. The control group received expectant management. RESULTS: The difference in the mean interval from pPROM to delivery between the groups was not statistically significant. Neonatal and maternal outcomes were significantly improved in the study group compared with the control group (fetal distress [10% vs 37%]; early neonatal sepsis [17% vs 63%]; neonatal mortality [17% vs 63%]; spontaneous delivery [83% vs 53%]; and postpartum sepsis [7% vs 33%]). CONCLUSION: Transabdominal amnioinfusion reduced fetal distress, early neonatal sepsis, and neonatal mortality. In the study group, more participants delivered spontaneously and there were fewer cases of postpartum sepsis, although the pPROM-delivery interval was not increased. PMID- 20015491 TI - Urinary incontinence in hospital-based nurses working in Turkey. AB - OBJECTIVE: To determine the changes in quality of life due to urinary incontinence in female hospital-based nurses working in Turkey and whether professional help was sought by the nurses. Another aim was to determine the prevalence of urinary incontinence and its risk factors in working nurses. METHODS: Data were collected through administration of a questionnaire and the Incontinence Quality-of-Life Instrument. The data were analyzed via chi(2) test, t test, one-way analysis of variance, and binary logistic regression analysis. RESULTS: The prevalence of urinary incontinence was 21.5%. Explanatory variables for developing urinary incontinence were age, parity, and recurrent urinary tract infections. Nurses with stress incontinence or urge incontinence had better quality of life than did those with mixed incontinence. CONCLUSION: In the present study, urinary incontinence was common in working nurses, and had a negative impact on their general quality of life. PMID- 20015493 TI - An imbalanced OPG/TRAIL ratio is associated to severe acute myocardial infarction. AB - BACKGROUND: This study was designed to evaluate the biological significance of simultaneous changes in the circulating levels of osteoprotegerin (OPG) and TNF related apoptosis inducing ligand (TRAIL) in patients with coronary artery disease (CAD), and, in particular, with acute myocardial infarction (AMI). METHODS: Total levels of OPG and TRAIL were measured by ELISA in patients with AMI (n=113), unstable angina (UA, n=21) and healthy controls (n=120). RESULTS: Since OPG was elevated during the acute phase (first 12-24-48h) after AMI and in patients with UA with respect to healthy controls, while TRAIL was decreased in acute AMI patients, CAD patients were characterized by an increased OPG/TRAIL ratio. Moreover, the OPG/TRAIL ratio was significantly (p<0.05) higher in the acute AMI patients who developed heart failure (HF) than in those who did not develop HF in the follow-up. CONCLUSIONS: An impaired OPG/TRAIL ratio after AMI is related to a higher risk of HF. PMID- 20015492 TI - The association of uncarboxylated matrix Gla protein with mitral annular calcification differs by diabetes status: The Heart and Soul study. AB - OBJECTIVE: Mitral annular calcification (MAC) and aortic stenosis (AS) are associated with systemic calcification and cardiovascular disease (CVD) events. Matrix Gla protein (MGP) is an inhibitor of vascular calcification and lower levels of its precursor - uncarboxylated MGP (ucMGP) - are associated with vascular calcification in pilot studies. METHODS AND RESULTS: In this cross sectional study of 839 outpatients with stable CVD, we measured serum ucMGP, and evaluated MAC and AS by echocardiography. The association of ucMGP with MAC differed by diabetes status (interaction P<0.001). Among participants without diabetes (n=615), higher ucMGP (per standard deviation [1178nM] increase) was associated with lower odds of MAC (odds ratio [OR] 0.73; 95% confidence interval [CI] 0.55-0.97) in models adjusted for traditional CVD risk factors, C-reactive protein, and kidney function. Among persons with diabetes (n=221), higher ucMGP was associated with higher odds of MAC (OR 1.89; 95% CI 1.29-2.78). Results were qualitatively similar for the association of ucMGP with AS although not statistically significant. CONCLUSIONS: Among outpatients with stable CVD, higher ucMGP is associated with lower odds of MAC in persons without diabetes, and higher odds of MAC in persons with diabetes. Future studies should determine whether ucMGP levels are associated with CVD events, and whether such associations differ by diabetes status. PMID- 20015494 TI - Effect of acetabular labral tears, repair and resection on hip cartilage strain: A 7T MR study. AB - BACKGROUND: Tears of the acetabular labrum are frequently present in patients with groin pain. While it is clear that the labrum contributes to the surface area articulating with the femoral head, it is not clear whether labral repair yields different load distribution in the hip compared to labral resection. PURPOSE: Determine whether labral repair reduces cartilage strain more effectively than labral resection. METHODS: Six human cadaveric hips (mean age 37 years) were loaded in a simulated single-leg stance within the bore of a 7T MR scanner. After cartilage had reached a steady-state thickness distribution, a scan of the cartilage was acquired with a voxel size of 0.1x0.1x0.3mm. This method was repeated for each of six specimens when the labrum was intact, after a surgically simulated labral tear, after an arthroscopic labral repair and after labral resection. Cartilage thickness and strain in an anterosuperior region of interest were measured from the MR scans. A paired t-test was used to compare mean and maximum cartilage strain when the labrum was intact vs. torn, torn vs. repaired and repaired vs. resected. Three-dimensional patterns of cartilage strain distribution were qualitatively compared for the different labral conditions. RESULTS: For the number of specimens tested we found no change in mean and maximum cartilage strain, and little obvious change in the pattern of cartilage strain distribution after a simulated labral tear. Labral repair caused a 2% decrease in mean cartilage strain compared to a torn labrum (p=0.014). Labral resection caused a 4% and 6% increase in mean and maximum cartilage strain, respectively, compared to labral repair (p=0.02), and the cartilage strain distribution was elevated throughout the region of interest. CONCLUSION: Based on our ex vivo findings of increased cartilage strain after labral resection when compared to labral repair, we have demonstrated the associated consequences to the mechanical environment of the cartilage following surgical treatment of the labrum. PMID- 20015495 TI - Joint moment contributions to swing knee extension acceleration during gait in children with spastic hemiplegic cerebral palsy. AB - Inadequate peak knee extension during the swing phase of gait is a major deficit in individuals with spastic cerebral palsy (CP). The biomechanical mechanisms responsible for knee extension have not been thoroughly examined in CP. The purpose of this study was to assess the contributions of joint moments and gravity to knee extension acceleration during swing in children with spastic hemiplegic CP. Six children with spastic hemiplegic CP were recruited (age=13.4+/ 4.8 years). Gait data were collected using an eight-camera system. Induced acceleration analysis was performed for each limb during swing. Average joint moment and gravity contributions to swing knee extension acceleration were calculated. Total swing and stance joint moment contributions were compared between the hemiplegic and non-hemiplegic limbs using paired t-tests (p<0.05). Swing limb joint moment contributions from the hemiplegic limb decelerated swing knee extension significantly more than those of the non-hemiplegic limb and resulted in significantly reduced knee extension acceleration. Total stance limb joint moment contributions were not statistically different. Swing limb joint moment contributions that decelerated knee extension appeared to be the primary cause of inadequate knee extension acceleration during swing. Stance limb muscle strength did not appear to be the limiting factor in achieving adequate knee extension in children with CP. Recent research has shown that the ability to extend the knee during swing is dependent on the selective voluntary motor control of the limb. Data from individual participants support this concept. PMID- 20015496 TI - Separation and preconcentration of persistent organic pollutants by cloud point extraction. AB - Persistent organic pollutants (POPs) are recognized as a class of poisonous compounds which pose risks of causing adverse effects to human health and the environment. Thus, it is very important to detect POPs in environmental and biological samples. The identification and determination of very low levels of POPs in complex matrices is extremely difficult. Recently a promising environmentally benign extraction and preconcentration methodology based on cloud point extraction (CPE) has emerged as an efficient sample pretreatment technique for the determination of trace/ultra-trace POPs in complex matrices. The purpose of this paper is to review the past and latest use of CPE for preconcentrating POPs and its coupling to different contemporary instrumental methods of analysis. First, the comparison of various extraction techniques for POPs is described. Next, the general concept, influence factors and other methods associated with CPE technique are outlined and described. Last, the hyphenations of CPE to various instrumental methods for their determination are summarized and discussed. PMID- 20015497 TI - Trace determination of sulphur mustard and related compounds in water by headspace-trap gas chromatography-mass spectrometry. AB - A method for trace determination of sulphur mustard (HD) and some of its cyclic decomposition compounds in water samples has been developed using headspace-trap in combination with gas chromatography-mass spectrometry (GC-MS). Factorial design was used for optimisation of the method. The trap technology allows enrichment and focusing of the analytes on an adsorbent, hence the technique offers better sensitivity compared to conventional static headspace. A detection limit of 1ng/ml was achieved for HD, while the cyclic sulphur compounds 1,4 thioxane, 1,3-dithiolane and 1,4-dithiane could be detected at a level of 0.1ng/ml. The method was validated for the stable cyclic compounds in the concentration range from the limit of quantification (LOQ: 0.2-0.4ng/ml) to hundred times LOQ. The within and between assay precisions at hundred times LOQ were 1-2% and 7-8% relative standard deviation, respectively. This technique requires almost no sample handling, and the total time for sampling and analysis was less than 1h. The method was successfully employed for muddy river water and sea water samples. PMID- 20015498 TI - Further investigations of the effect of pressure on retention in ultra-high pressure liquid chromatography. AB - In this study, we investigated further the large increases in retention with pressure that we observed previously in RP-LC especially for ionised solutes. These findings were initially confirmed on a conventional silica C(18) column, which gave extremely similar results to the hybrid C(18) phase originally used. Large increases in retention factor of approximately 50% for a pressure increase of 500 bar were also shown for high MW polar but neutral solutes. However, experiments with the same bases in ionised and non-ionised forms suggest that somewhat greater pressure-induced retention increases are found for ionised solutes. Retention increases with pressure were found to be considerably smaller for a C(1) column compared with a C(18) column; decreases in retention with increasing pressure were noted for ionised bases when using a bare silica column in the hydrophilic interaction chromatography (HILIC) mode. These observations are consistent with the partial loss of the solvation layer in RP-LC as the solute is forced into the hydrophobic environment of the stationary phase, and consequent reduction in the solute molar volume, while the water layer on the surface of a HILIC packing increases the hydration of a basic analyte. Finally, retention changes with pressure in RP-LC can also be observed at a mobile phase pH close to the solute pK(a), due to changes in pK(a) with pressure. However, this effect has no influence on the results of most of our studies. PMID- 20015499 TI - Development of the custom polymeric materials specific for aflatoxin B1 and ochratoxin A for application with the ToxiQuant T1 sensor tool. AB - Two polymers were computationally designed with affinity to two of the most abundant mycotoxins aflatoxin B1 (AFB1) and ochratoxin A (OTA) for application in the ToxiQuant T1 System. The principle of quantification of AFB1 and OTA using the ToxiQuant T1 instrument comprised of a fluorimetric analysis of mycotoxins adsorbed on the polymer upon exposure to UV light. High affinity of the developed resins allowed the adsorption of both toxins as discrete bands on the top of the cartridge with detection limit as low as 1ng quantity of mycotoxins. PMID- 20015500 TI - High-efficiency hydrophilic interaction chromatography by coupling 25 cm x 4.6mm ID x 5 microm silica columns and operation at 80 degrees C. AB - Recently, hydrophilic interaction chromatography (HILIC) has emerged as a valuable orthogonal tool to reversed-phase liquid chromatography (RP-LC) as it allows for resolution of highly polar ionisable compounds. The relationships between separation efficiency, column length and speed of analysis for 4.6 mm ID x 5 microm silica particle columns in HILIC are demonstrated using kinetic plots. The kinetic plots constructed for conventional pressure systems operating at 350 bar and at 30 degrees C and 80 degrees C are confirmed using experimental data for different column lengths. Efficiencies of more than 130,000 theoretical plates could be achieved by connecting up to six columns of 25 cm. As expected, a significant gain in analysis speed without loss of efficiency could be obtained by operating at 80 degrees C compared to 30 degrees C. The advantages of using long columns in HILIC in combination with elevated column temperature for the pharmaceutical industry are illustrated using test mixtures comprised of commercially available ionisable compounds (including some containing functional groups with potential genotoxic typical structural alerts) as well as real polar ionisable pharmaceuticals. PMID- 20015501 TI - Multi-wall carbon nanotubes bonding on silica-hydride surfaces for open-tubular capillary electrochromatography. AB - Prepared multi-wall carbon nanotube (MWNT) materials, including untreated MWNT, HNO(3)-treated MWNT and HNO(3)-HCl-treated MWNT were covalently attached onto a silica-hydride-modified capillary by hydrosilation, using the abundant double bonds between the pentagon carbons in the MWNT structure. These MWNT-incorporated capillaries were characterized by SEM, ATR-IR and electroosmotic flow (EOF) measurements in phosphate buffers with a pH range of 3.7-9.3 and in the mixtures of acetonitrile modifier. The untreated capillary was assumed to carry some carboxylate groups formed on the non-acid-treated MWNTs, as it had higher EOF values than the hydride capillary. As the MWNTs were treated with HNO(3) and HCl solutions, the capillaries had increasingly higher EOF values. To examine the existence of an electrochromatography mechanism in the modified capillaries, a mixture of nucleosides and thymine was probed to check the velocity factor and retention factor. In addition to the pi-pi interaction between the probe solutes and the MWNT immobilized stationary phases; a reversed-phase mechanism could contribute to the chromatographic retention. For acidic tetracyclines, increasing the loadability of MWNTs resulted in a high retention factor and improved the separation resolution. PMID- 20015502 TI - Profiling analysis of volatile compounds from fruits using comprehensive two dimensional gas chromatography and image processing techniques. AB - An image processing approach originating from the proteomics field has been transferred successfully to the processing of data obtained with comprehensive two-dimensional gas chromatographic separations data. The approach described here has proven to be a useful analytical tool for unbiased pattern comparison or profiling analyses, as demonstrated with the differentiation of volatile patterns ("aroma") from fruits such as apples, pears, and quince fruit. These volatile patterns were generated by headspace solid phase microextraction coupled to comprehensive two-dimensional gas chromatography (HS-SPME-GC x GC). The data obtained from GC x GC chromatograms were used as contour plots which were then converted to gray-scale images and analyzed utilizing a workflow derived from 2D gel-based proteomics. Run-to-run variations between GC x GC chromatograms, respectively their contour plots, have been compensated by image warping. The GC x GC images were then merged into a fusion image yielding a defined and project wide spot (peak) consensus pattern. Within detected spot boundaries of this consensus pattern, relative quantities of the volatiles from each GC x GC image have been calculated, resulting in more than 700 gap free volatile profiles over all samples. These profiles have been used for multivariate statistical analysis and allowed clustering of comparable sample origins and prediction of unknown samples. At present state of development, the advantage of using mass spectrometric detection can only be realized by data processing off-line from the identified software packages. However, such information provides a substantial basis for identification of statistically relevant compounds or for a targeted analysis. PMID- 20015503 TI - Rapid simultaneous determination of dexamethasone and betamethasone in milk by liquid chromatography tandem mass spectrometry with isotope dilution. AB - A simple, sensitive and reliable analytical method for the rapid simultaneous determination of dexamethasone and betamethasone in milk by high performance liquid chromatography-negative electrospray ionization tandem mass spectrometry (HPLC-NESI-MS/MS) with isotope dilution was developed. Samples were directly purified through C(18) cartridge. Then the eluate was dried under nitrogen and residues were dissolved in mobile phase. Samples were analyzed by HPLC-MS/MS on a Hypercarb graphite column with a mixture of acetonitrile-water-formic acid as mobile phase. The samples were quantified using dexamethasone-D(4) as an internal standard. The procedure was validated according to the European Union regulation 2002/657/EC determining specificity, decision limit (CCalpha), detection capability (CCbeta), trueness, precision, linearity and stability. The method is demonstrated to be suitable for the determination of dexamethasone and betamethasone in milk. The total time required for the analysis of one sample was about 35 min. PMID- 20015504 TI - Strategies for the on-line preconcentration and separation of hypolipidaemic drugs using micellar electrokinetic chromatography. AB - Three strategies were investigated for the simultaneous separation and on-line preconcentration of charged and neutral hypolipidaemic drugs in micellar electrokinetic chromatography (MEKC). A background electrolyte (BGE) consisting of 20 mM ammonium bicarbonate buffer (pH 8.50) and 50 mM sodium dodecyl sulfate (SDS) was used for the separation and on-line preconcentration of the drugs. The efficiencies of sweeping, analyte focusing by micelle collapse (AFMC), and simultaneous field-amplified sample stacking (FASS) and sweeping, were compared for the preconcentration of eight hypolipidaemic drugs in different conductivity sample matrices. When compared with a hydrodynamic injection (5 s at 50 mbar, 0.51% of capillary volume to detection window) of drug mixture prepared in the separation BGE, improvements of detection sensitivity of 60-, 83-, and 80-fold were obtained with sweeping, AFMC and simultaneous FASS and sweeping, respectively, giving limits of detection (LODs) of 50, 36, and 38 microg/L, respectively. The studied techniques showed suitability for focusing different types of analytes having different values of retention factor (k). This is the first report for the separation of different types of hypolipidaemic drugs by capillary electrophoresis (CE). The three methods were validated then applied for the analysis of target analytes in wastewater samples from Hobart city. PMID- 20015505 TI - Dynamic interconversion of chiral oxime compounds in gas chromatography. AB - A number of chiral oxime compounds have been synthesised and their gas chromatographic analysis on both a polyethelene glycol phase column and two chiral column phases was investigated. Of particular interest to this work is the observation of dynamic interconversion behaviour, both in a single dimensional analysis, and by using comprehensive two-dimensional gas chromatography (GCxGC). A number of non-chiral compounds were studied as a means to understand the nature of the behaviour observed. As expected, the achiral compound on both the wax column and the chiral column generated two isomeric compounds-the E and Z isomers. On the wax column, a characteristic interconversion zone representing the dynamic process was observed, with extent of interconversion dependent on the conditions used. For the chiral compounds, two isomers and the interconversion zone were exhibited on the wax column, however on the chiral column 4 isomeric peaks were found-the (R) and (S) enantiomers of each of the E and Z isomers. In the case of the chiral column, the extent of interconversion was negligible, and this appears to correlate with the use of low polarity columns. In order to encourage dynamic interconversion, a polyethylene glycol column was coupled to the chiral column, by placing it either before or after the chiral column. In this case a monitor detector was employed between the two columns in order to isolate the effects of the first column from the behaviour on the second. In a further study, the most appropriate column arrangement from the earlier study was placed into a comprehensive two-dimensional gas chromatography instrument, with a wax-phase column in the second dimension. The unique location of peaks for each of the molecules in 2D space and patterns for the interconversion processes is interpreted phenomenologically. PMID- 20015506 TI - Thermo-responsive polymer brush-grafted porous polystyrene beads for all-aqueous chromatography. AB - Poly(N-isopropylacrylamide) (PIPAAm) brush-grafted porous polystyrene beads with variable grafted polymer densities were prepared using surface-initiated atom transfer radical polymerization (ATRP) for applications in thermo-responsive chromatography. Utilization of these grafted beads as a stationary phase in aqueous chromatographic analysis of insulin provides a graft density-dependent analyte retention behavior. The separations calibration curve on PIPAAm-grafted polystyrene was obtained using pullulan standards and exhibited inflection points attributed to analyte diffusion into bead pores and partitioning into grafted PIPAAm brush surfaces. Presence of these inflection points supports a separation mechanism where insulin penetrates pores in polystyrene beads and hydrophobically interacts with PIPAAm brushes grafted within the pores. Control of PIPAAm brush graft density on polystyrene facilitates effective aqueous phase separation of peptides based on thermally modulated hydrophobic interactions with grafted PIPAAm within stationary phase pores. These results indicated that PIPAAm brush grafted porous polystyrene beads prepared by surface-initiated ATRP was effective stationary phase of thermo-responsive chromatography for aqueous phase peptide separations. PMID- 20015507 TI - On- and off column enantiomerization of 4,4'-bisquinolin-2-ones: A comparison of Auto-, DHPLcy2k and DCXplorer calculated thermodynamic data generated by dynamic high, performance liquid chromatography with theoretically calculated data. AB - Fast semipreparative HPLC enantioseparation of four axially chiral biscarbostyrils (4,4'-bisquinoline-2-ones) using ULMO as a pi-acidic Pirkle type chiral stationary phase leads to two racemizing pairs (1,2; k(obs) 1.6x10(-4) and 3.0x10(-4)s(-1) at 28 degrees C) and two stable ones (3,4). 3 was stabilized by a crown ether linkage from pos. 6 to 6', and 4 had sterically demanding bromo substituents in pos. 3 and 3'. On-column generated temperature-dependent chromatograms of 1 and 2 were fitted with Auto-DHPLCy2k and DCXplorer. For cpd 2 both programs delivered similar DeltaG values of 90 and 93kJ/mol, well comparable with the 99kJ/mol calculated with the B3LYP/6-31G (d) procedure. At temperatures of high conversion DCXplorer delivered inconsistent series of rate constants for the more tailing and less resolved tetramethoxy derivative 1. We connect this problem with an almost impossible halfwidth calculation of tailing peak pairs which are weakly resolved. However, this problem could be observed only in the case of tetramethoxy derivative 1. Stochastic generated data of Auto-DHPLCy2k could be used at a lower percentage of conversion only while the theoretical plate model did not deliver useful data at temperatures of very low conversion but fitted well high conversion chromatogram series of 1 and 2. PMID- 20015508 TI - Liquid chromatography/mass spectrometry analysis of perfluoroalkyl carboxylic acids and perfluorooctanesulfonate in bivalve shells: extraction method optimization. AB - Different extraction methods, including extraction by organic solvents with and without acetic acid digestion, and mixed inorganic acid digestion coupled with solid phase extraction (SPE), were developed for the analysis of perfluorinated carboxylic acids (PFCAs) and perfluorooctanesulfonate (PFOS) in bivalve shells. The extracts were separated, identified and quantified by liquid chromatography electrospray ionization-tandem mass spectrometry (LC-ESI-MS/MS). The method utilizing mixed acid digestion coupled with SPE performed more efficiently than other extraction methods. Matrix recoveries of the optimized methods ranged from 92% to 104%, with limits of detection of 0.05-0.43ng/g. The optimized method was successfully applied to the analysis of PFCAs and PFOS in shell samples of two bivalves from Bohai Bay, China. PFCAs and PFOS concentrations in the shells ranged from 0.3 ng/g to 4.1 ng/g, 1-50 times lower than those in the soft tissues of bivalves for most target analytes. No relationship between PFCAs and PFOS in shells and in soft tissues was found; this is explained by the different contaminant uptake mechanism of shells and soft tissues. PMID- 20015509 TI - Gas chromatographic-mass spectrometric analysis of volatiles obtained by four different techniques from Salvia rosifolia Sm., and evaluation for biological activity. AB - Four different isolation techniques, conventional hydrodistillation (HD), microwave-assisted hydrodistillation (MWHD), microdistillation (MD) and micro steam distillation-solid-phase microextraction (MSD-SPME), have been used to analyze the volatile constituents from the aerial parts of Salvia rosifolia Sm. by gas chromatography and gas chromatography coupled to mass spectrometry. HD and MWHD techniques produced quantitatively (yield, 0.39% and 0.40%) and qualitatively (aromatic profile) similar essential oils. alpha-Pinene (15.7 34.8%), 1,8-cineole (16.6-25.1%), beta-pinene (6.7-13.5%), beta-caryophyllene (1.4-5.0%) and caryophyllene oxide (1.4-4.4%) were identified as major constituents of this Turkish endemic species. Besides, the hydrodistilled oil of S. rosifolia was evaluated for antibacterial, antifungal, anticancer, antioxidant and cytotoxic activities. The hydrodistilled oil of S. rosifolia showed antibacterial activity against Methicillin-resistant Staphylococcus aureus (MRSA) with a MIC value of 125microg/mL. Other human pathogenic microorganisms (Escherichia coli, Pseudomonas aeruginosa, Enterobacter aerogenes, Salmonella typhimurium, Staphylococcus epidermidis, Candida albicans) were also inhibited within a moderate range (MIC=125-1000microg/mL). Antifungal activity of the oil was also observed against the strawberry anthracnose-causing fungal plant pathogens Colletotrichum acutatum, C. fragariae and C. gloeosporioides. No cytotoxicity was observed for S. rosifolia oil up to 25mg/mL against malignant melanoma, epidermal, ductal and ovary carcinoma. PMID- 20015510 TI - Trace determination of pharmaceuticals and other wastewater-derived micropollutants by solid phase extraction and gas chromatography/mass spectrometry. AB - The presence of pharmaceuticals and other wastewater-derived micropollutants in surface and groundwaters is receiving intense public and scientific attention. Yet simple GC/MS methods that would enable measurement of a wide range of such compounds are scarce. This paper describes a GC/MS method for the simultaneous determination of 13 pharmaceuticals (acetaminophen, albuterol, allopurinol, amitriptyline, brompheniramine, carbamazepine, carisoprodol, ciclopirox, diazepam, fenofibrate, metoprolol, primidone, and terbinafine) and 5 wastewater derived contaminants (caffeine, diethyltoluamide, n-butylbenzene sulfonamide, n nonylphenol, and n-octylphenol) by solid phase extraction (SPE) and derivatization with BSTFA. The method was applied to the analysis of raw and treated sewage samples obtained from a wastewater treatment plant located in the mid-Atlantic United States. All analytes were detected in untreated sewage, and 14 of the 18 analytes were detected in treated sewage. PMID- 20015511 TI - Introducing porous graphitized carbon liquid chromatography with evaporative light scattering and mass spectrometry detection into cell wall oligosaccharide analysis. AB - Separation and characterization of complex mixtures of oligosaccharides is quite difficult and, depending on elution conditions, structural information is often lost. Therefore, the use of a porous-graphitized-carbon (PGC)-HPLC-ELSD-MS(n) method as analytical tool for the analysis of oligosaccharides derived from plant cell wall polysaccharides has been investigated. It is demonstrated that PGC-HPLC can be widely used for neutral and acidic oligosaccharides derived from cell wall polysaccharides. Furthermore, it is a non-modifying technique that enables the characterization of cell wall oligosaccharides carrying, e.g. acetyl groups and methylesters. Neutral oligosaccharides are separated based on their size as well as on their type of linkage and resulting 3D-structure. Series of the planar beta (1,4)-xylo- and beta-(1,4)-gluco-oligosaccharides are retained much more by the PGC material than the series of beta-(1,4)-galacto-, beta-(1,4)-manno- and alpha (1,4)-gluco-oligosaccharides. Charged oligomers such as alpha-(1,4)-galacturonic acid oligosaccharides are strongly retained and are eluted only after addition of trifluoroacetic acid depending on their net charge. Online-MS-coupling using a 1:1 splitter enables quantitative detection of ELSD as well as simple identification of many oligosaccharides, even when separation of oligosaccharides within a complex mixture is not complete. Consequently, PGC-HPLC-separation in combination with MS-detection gives a powerful tool to identify a wide range of neutral and acidic oligosaccharides derived from various cell wall polysaccharides. PMID- 20015512 TI - Determination of tetracyclines in surface water and milk by the magnesium hydroxide coprecipitation method. AB - A simple coprecipitation method was developed for the determination of tetracyclines (TCs) in surface water and milk by high-performance liquid chromatography with diode-array detection (HPLC-DAD). Magnesium ion was added into the surface water or the acetonitrile (MeCN) extract of milk. After alkalinization, magnesium hydroxide precipitates which had been formed can be separated from the matrix solution easily by centrifuging and then a dissolution step was performed by adding a small amount of acid. The final solution could be introduced directly into HPLC system for the determination of the analytes. Under optimal conditions, recoveries for the analysis of spiked surface water samples ranged from 83.6% to 95.1% with relative standard deviation of 2.0-5.5%. For milk samples, relative recoveries were 95.9-104.6% with relative standard deviation of 3.4-6.7%. The enrichment factors ranged from 41.5 to 48.1 for 10 mL water samples, and from 3.6 to 4.4 for 1 mL MeCN extracts of milk. Limits of detection ranged from 0.13 to 0.51 ng/mL, and from 3.0 to 8.5 ng/g for four TCs in surface water and milk samples, respectively. PMID- 20015513 TI - Small angle scattering model for Pickering emulsions and raspberry particles. AB - Pickering emulsions, raspberry particles and other colloidal particle complexes are often characterized using small angle scattering techniques. The present work derives an analytical scattering model that accounts for the self-correlation of a spherical core and surface adsorbed particles as well as the particle-particle and core-particle correlation terms characteristic of Pickering emulsions and raspberry particles. It is shown that contrast matching of the scattering length density is not essential to obtain meaningful information as long as the scattering contrasts of all phases are precisely known. The derived equations are useful for analyzing data and planning experiments for Small Angle Neutron Scattering (SANS) and Small Angle X-ray Scattering (SAXS) involving these colloidal systems. PMID- 20015514 TI - Infants' recognition of objects using canonical color. AB - We explored infants' ability to recognize the canonical colors of daily objects, including two color-specific objects (human face and fruit) and a non-color specific object (flower), by using a preferential looking technique. A total of 58 infants between 5 and 8 months of age were tested with a stimulus composed of two color pictures of an object placed side by side: a correctly colored picture (e.g., red strawberry) and an inappropriately colored picture (e.g., green-blue strawberry). The results showed that, overall, the 6- to 8-month-olds showed preference for the correctly colored pictures for color-specific objects, whereas they did not show preference for the correctly colored pictures for the non-color specific object. The 5-month-olds showed no significant preference for the correctly colored pictures for all object conditions. These findings imply that the recognition of canonical color for objects emerges at 6 months of age. PMID- 20015516 TI - The combination of elevated BNP and AF as a predictor of no early recanalization after IV-t-PA in acute ischemic stroke. AB - BACKGROUND AND PURPOSE: In acute stroke patients treated with intravenous tissue plasminogen activator (t-PA), early recanalization can improve patient outcome. Heart failure may result in reduction of brain perfusion, which limits the ability of the blood stream to wash out emboli. Brain natriuretic peptide (BNP) is used as a biological marker of heart failure. Most stroke patients with atrial fibrillation (AF) have elevated BNP levels. We investigated the relationships of plasma BNP levels before t-PA infusion and AF with early recanalization after t PA infusion. METHODS: Patients with a major brain artery occlusion were studied prospectively. MRAs were performed before and within 60 min after t-PA infusion. The relationship between BNP levels before t-PA infusion and the presence of AF with early recanalization was examined. RESULTS: Seventy-nine patients (49 men; mean age, 75.5+/-10.4 years; ICA occlusion in 25 patients, M1 in 32, M2 in 13, PCA in 3, and BA in 6) were enrolled. Follow-up MRA within 60 min after t-PA infusion revealed recanalization in 35 (44.3%) patients and no recanalization in 44 (55.7%). Patients with AF (57.1% vs. 75.0%, P=0.0294) and BNP>150pg/dl (39.0% vs. 73.7%, P=0.0019) less frequently had early recanalization than those without AF and with BNP< or =150pg/dl. The combination of AF and BNP>150pg/ml was a useful predictor for no early recanalization (positive predictive value, 79.4%; negative predictive value, 62.2%; sensitivity, 61.4%; specificity, 80.0%). CONCLUSION: The presence of AF and elevated BNP was associated with no early recanalization after IV-t-PA therapy. We should need further study to ascertain its predictive ability. PMID- 20015515 TI - Selective alterations of the CB1 receptors and the fatty acid amide hydrolase in the ventral striatum of alcoholics and suicides. AB - Recent studies in rodents have suggested a role for the central endocannabinoid system in the regulation of mood and alcohol related behaviors. Alcohol use disorder is often associated with suicidal behavior. In the present study, we examined whether abnormalities in the endocannabinoid system in the ventral striatum are associated with alcohol dependence and suicide. The levels of CB1 receptors, receptor-mediated G-protein signaling, and activity and level of the fatty acid amide hydrolase (FAAH) were analyzed postmortem in the ventral striatum of alcohol-dependent nonsuicides (CA, n=9), alcohol-dependent suicides (AS, n=9) and nonpsychiatric controls (C, n=9). All subjects underwent a psychological autopsy, and toxicological and neuropathological examinations. The levels of the CB1 receptors and the CB1 receptor-mediated G-protein signaling were significantly lower in the ventral striatum of CA compared to the control group. However, these parameters were elevated in AS when compared to CA group. The activity of FAAH enzyme was lower in CA compared to the control group while it was found to be significantly higher in AS compared with CA group. These findings suggest that alcohol dependence is associated with the downregulation of the CB1 receptors, while suicide is linked to the upregulation of these receptors in the ventral striatum. Alteration in the activity of FAAH enzyme that regulates the anandamide (AEA) content might in turn explain differences in the CB1 receptor function in alcohol dependence and suicide. These findings may have etiological and therapeutic implications for the treatment of alcohol addiction and suicidal behavior. PMID- 20015517 TI - Microbiological water quality at non-human influenced reference beaches in southern California during wet weather. AB - Although urban wet weather discharges may have elevated concentrations of fecal indicator bacteria impacting water quality at swimming beaches, not all of these bacteria may arise from human sources. In this study, the contribution of non human fecal indicator bacteria was quantified by sampling coastal reference beaches in southern California. Samples were collected at beaches near stormwater discharges from undeveloped watersheds and analyzed for total coliform, Escherichia coli, and enterococci. Surfzone samples exceeded water quality thresholds >10 times more frequently during wet weather than dry weather. Exceedences were greatest <24h following rainfall, then steadily declined on successive days. Early season storms exceeded thresholds more frequently, and by greater magnitude, compared to late season storms. Large storms exceeded thresholds more frequently than smaller-sized storms, partly due to the breaching of sand berms. When discharges did reach the surf zone, bacterial concentrations in the wave wash were correlated with watershed bacterial flux. PMID- 20015518 TI - alpha-Lipoic acid increases energy expenditure by enhancing adenosine monophosphate-activated protein kinase-peroxisome proliferator-activated receptor gamma coactivator-1alpha signaling in the skeletal muscle of aged mice. AB - Skeletal muscle mitochondrial dysfunction is associated with aging and diabetes, which decreases respiratory capacity and increases reactive oxygen species. Lipoic acid (LA) possesses antioxidative and antidiabetic properties. Metabolic action of LA is mediated by activation of adenosine monophosphate-activated protein kinase (AMPK), a cellular energy sensor that can regulate peroxisome proliferator-activated receptor-gamma coactivator-1alpha (PGC-1alpha), a master regulator of mitochondrial biogenesis. We hypothesized that LA improves energy metabolism and mitochondrial biogenesis by enhancing AMPK-PGC-1alpha signaling in the skeletal muscle of aged mice. C57BL/6 mice (24 months old, male) were supplemented with or without alpha-LA (0.75% in drinking water) for 1 month. In addition, metabolic action and cellular signaling of LA were studied in cultured mouse myoblastoma C2C12 cells. Lipoic acid supplementation improved body composition, glucose tolerance, and energy expenditure in the aged mice. Lipoic acid increased skeletal muscle mitochondrial biogenesis with increased phosphorylation of AMPK and messenger RNA expression of PGC-1alpha and glucose transporter-4. Besides body fat mass, LA decreased lean mass and attenuated phosphorylation of mammalian target of rapamycin (mTOR) signaling in the skeletal muscle. In cultured C2C12 cells, LA increased glucose uptake and palmitate beta oxidation, but decreased protein synthesis, which was associated with increased phosphorylation of AMPK and expression of PGC-1alpha and glucose transporter-4, and attenuated phosphorylation of mTOR and p70S6 kinase. We conclude that LA improves skeletal muscle energy metabolism in the aged mouse possibly through enhancing AMPK-PGC-1alpha-mediated mitochondrial biogenesis and function. Moreover, LA increases lean mass loss possibly by suppressing protein synthesis in the skeletal muscle by down-regulating the mTOR signaling pathway. Thus, LA may be a promising supplement for treatment of obesity and/or insulin resistance in older patients. PMID- 20015519 TI - Calcium deficiency in the early stages after weaning is associated with the enhancement of a low level of adrenaline-stimulated lipolysis and reduction of adiponectin release in isolated rat mesenteric adipocytes. AB - Dysregulation of visceral adipocytes increases the incidence of metabolic syndrome. Higher production of nonesterified fatty acid and changes in adipocytokine release may trigger insulin resistance. Many studies have suggested that calcium (Ca) deficiency is associated with insulin resistance; however, the mechanisms are poorly understood. We examined the effects of Ca deficiency on adrenaline-induced lipolysis and adipocytokine release in the early stages after weaning using freshly isolated adipocytes from mesenteric fat tissue of 3-week old male Sprague-Dawley rats fed a normal-Ca (5 g/kg diet) or low-Ca (1 g/kg diet) diet for 4 weeks. The release rate of nonesterified fatty acid in the mesenteric adipocytes after stimulation with a low level of adrenaline (0.2 microg/mL) was much higher in the Ca-deficient group than in the control group. In contrast, adiponectin release in the mesenteric fat cells was lower in Ca deficient rats. Leptin and tumor necrosis factor-alpha secretion showed a similar tendency without significant intergroup differences, and monocyte chemoattractant protein-1 release was not affected by Ca deficiency. We found that Ca deficiency reduced the average size of fat cells through a large increase in the number of cells slightly smaller than the average size, which may be associated with the changes in the properties of the mesenteric adipose tissue. Our present results suggest that a low intake of Ca in the early stages after weaning is associated with changes in the properties of mesenteric adipocytes, which may be linked to insulin resistance in the future. PMID- 20015520 TI - Urgency of reassessment of role of obesity indices for metabolic risks. AB - The definition of metabolic syndrome places emphasis on health care for persons at risk. However, whether an obesity index should be a mandatory component of the definition and whether obesity indices can identify metabolic risks satisfactorily require further exploration. Therefore, we investigated the effectiveness of various anthropometric obesity indices in identifying the clustering of 2 or more American Heart Association (AHA)/National Heart, Lung, and Blood Institute (NHLBI)/International Diabetes Federation (IDF)-defined metabolic risk factors (hypertension, hyperglycemia, hypertriglyceridemia, and low high-density lipoprotein cholesterol) for metabolic syndrome and those of other metabolic risk factors (high low-density lipoprotein cholesterol, hyperuricemia, high gamma-glutamyltransferase, fatty liver) in 6141 men and 2137 women. The anthropometric indices were the following: (1) for both sexes--various levels of waist-to-height ratio (WHtR) including 0.5 and body mass index (BMI) of 23 and 25 kg/m(2); (2) for men and women individually--waist circumference (W) 90/80 cm (AHA/NHLBI/IDF for ethnic groups), W 85/90 cm (Japan Society for the Study of Obesity), and combined W and BMI: W 85/90 cm and/or BMI 25 kg/m(2) (Japanese government). The results showed the following: (1) The optimal value for WHtR was 0.5 for AHA/NHLBI/IDF-defined risk factors and approximately 0.5 for other risk factors in both sexes. (2) The sensitivities of various proposed obesity indices for identifying clustering of defined and other risk factors varied between 74.4% (WHtR 0.5) and 36.3% (BMI 25) and between 80.5% (WHtR 0.5) and 43.7% (BMI 25) in men, and varied between 65.6% (WHtR 0.5) and 16.8% (W 90 cm) and between 82.3% (WHtR 0.5) and 28.2% (W 90 cm) in women. Because the sensitivities of many anthropometric indices were very low, a reassessment of the effectiveness of obesity indices in evaluating metabolic risks and especially their suitability as a single mandatory component of metabolic syndrome is urgently needed. However, WHtR 0.5 provides a very useful algorithm for screening persons at risk. PMID- 20015521 TI - Cholesterol absorption decreases after Roux-en-Y gastric bypass but not after gastric banding. AB - The differences in cholesterol metabolism after the 2 most common forms of obesity surgery, Roux-en-Y gastric bypass (RYGB) and gastric banding (GB), have not been well characterized. In this study, effects of RYGB and GB on cholesterol absorption and synthesis were investigated. To this aim, 1-year follow-up of cholesterol metabolism in 2 nonrandomized cohorts undergoing either RYGB (n = 29; age, 45.2 +/- 7.7 years; body mass index [BMI], 46.0 +/- 6.1 kg/m(2)) or GB (n = 26; age, 45.9 +/- 8.6 years; BMI, 50.1 +/- 7.7 kg/m(2)) was performed in a university hospital center specializing in the treatment of morbid obesity. Serum markers of cholesterol synthesis (cholestenol, desmosterol, and lathosterol) and cholesterol absorption (campesterol, sitosterol, avenasterol, and cholestanol) were measured preoperatively and at follow-up and expressed as ratios to cholesterol. As expected based on observed weight loss (25% after RYGB and 17% after GB, P < .001 between groups), both operations decreased serum levels of cholesterol synthesis markers by 12% to 28% (all Ps < .001). A decrease in cholesterol absorption markers was only observed after RYGB (-26% for sitosterol) and not after GB (+16%, P = 2 x 10(-6) for difference between the groups). The difference in sitosterol ratio between the groups remained significant after adjustment for age, BMI, fasting insulin levels, and nutritional status (P = 2 x 10(-4)), indicating a specific effect related to RYGB. We conclude that decrease in cholesterol absorption is a novel beneficial effect of RYGB. Together with an improved control of blood glucose, this may contribute to a better cardiovascular risk profile after RYGB. PMID- 20015522 TI - Fetuin-A and atherosclerotic calcified plaque in patients with type 2 diabetes mellitus. AB - Fetuin-A is a multifunctional circulating glycoprotein. Among its roles, inhibition of ectopic calcification is a prominent feature. Low fetuin-A levels in dialysis patients are associated with cardiovascular mortality, possibly via accelerating vascular calcification. However, except for dialyzed conditions, a correlation between fetuin-A levels and vascular calcification remains controversial. Furthermore, any inhibitory effect of fetuin-A on atherosclerotic calcified plaques (CPs) remains unclear compared with its effect on medial artery calcification that is often found in dialyzed patients. Therefore, we examined the association between fetuin-A levels and atherosclerotic CPs. For this study, 416 consecutive patients with type 2 diabetes mellitus and without renal dysfunction were examined. We measured serum fetuin-A levels and investigated for the presence of CP in the common carotid and femoral arteries using ultrasonography. Fetuin-A levels were significantly lower in patients with CP than those without CP (262.6 +/- 56.7 and 281.5 +/- 64.6 microg/mL, respectively; P = .001). Multivariate logistic regression analysis showed that fetuin-A levels were inversely associated with the presence of CP (odds ratio = 0.753; 95% confidence interval, 0.608-0.933; P = .010). These results suggest that fetuin-A may inhibit the calcification of atherosclerotic plaques independently of the dialyzed condition. PMID- 20015523 TI - Preliminary report: leucine supplementation enhances glutamate dehydrogenase expression and restores glucose-induced insulin secretion in protein-malnourished rats. AB - Low-protein diet impairs insulin secretion in response to nutrients and may induce several metabolic disorders including diabetes, obesity, and cardiovascular disease. In the present study, the influence of leucine supplementation on glutamate dehydrogenase (GDH) expression and glucose-induced insulin secretion (GIIS) was investigated in malnourished rats. Four groups were fed with different diets for 12 weeks: a normal-protein diet (17%) without or with leucine supplementation or a low (6%)-protein diet without (LP) or with leucine supplementation (LPL). Leucine (1.5%) was supplied in the drinking water. Western blotting analysis revealed reduced GDH expression in LP, whereas LPL displayed improved GDH expression, similar to control. The GIIS and leucine induced insulin release were also enhanced in LPL compared with LP and similar to those observed in rats fed a normal-protein diet without leucine supplementation. In addition, GDH allosteric activators produced an increased insulin secretion in LPL. These findings indicate that leucine supplementation was able to increase GDH expression leading to GIIS restoration, probably by improved leucine metabolic pathways. PMID- 20015525 TI - Effects of sitagliptin or metformin added to pioglitazone monotherapy in poorly controlled type 2 diabetes mellitus patients. AB - The aim of the study was to compare the effects of the addition of sitagliptin or metformin to pioglitazone monotherapy in poorly controlled type 2 diabetes mellitus patients on body weight, glycemic control, beta-cell function, insulin resistance, and inflammatory state parameters. One hundred fifty-one patients with uncontrolled type 2 diabetes mellitus (glycated hemoglobin [HbA(1c)] >7.5%) in therapy with pioglitazone 30 mg/d were enrolled in this study. We randomized patients to take pioglitazone 30 mg plus sitagliptin 100 mg once a day, or pioglitazone 15 mg plus metformin 850 mg twice a day. We evaluated at baseline and after 3, 6, 9, and 12 months these parameters: body weight, body mass index, HbA(1c), fasting plasma glucose (FPG), postprandial plasma glucose (PPG), fasting plasma insulin (FPI), homeostasis model assessment insulin resistance index (HOMA IR), homeostasis model assessment beta-cell function index, fasting plasma proinsulin (Pr), Pr/FPI ratio, adiponectin, resistin (R), tumor necrosis factor alpha (TNF-alpha), and high-sensitivity C-reactive protein. A decrease of body weight and body mass index was observed with metformin, but not with sitagliptin, at the end of the study. We observed a comparable significant decrease of HbA(1c), FPG, and PPG and a significant increase of homeostasis model assessment beta-cell function index compared with baseline in both groups without any significant differences between the 2 groups. Fasting plasma insulin, fasting plasma Pr, Pr/FPI ratio, and HOMA-IR values were decreased in both groups even if the values obtained with metformin were significantly lower than the values obtained with sitagliptin. There were no significant variations of ADN, R, or TNF alpha with sitagliptin, whereas a significant increase of ADN and a significant decrease of R and TNF-alpha values were recorded with metformin. A significant decrease of high-sensitivity C-reactive protein value was obtained in both groups without any significant differences between the 2 groups. There was a significant correlation between HOMA-IR decrease and ADN increase, and between HOMA-IR decrease and R and TNF-alpha decrease in pioglitazone plus metformin group after the treatment. The addition of both sitagliptin or metformin to pioglitazone gave an improvement of HbA(1c), FPG, and PPG; but metformin led also to a decrease of body weight and to a faster and better improvement of insulin resistance and inflammatory state parameters, even if sitagliptin produced a better protection of beta-cell function. PMID- 20015524 TI - Exercise training improves cardiovascular autonomic modulation in response to glucose ingestion in obese adults with and without type 2 diabetes mellitus. AB - This study examined the effect of aerobic exercise training on vagal and sympathetic influences on the modulations of heart rate and systolic blood pressure in response to an oral glucose load in obese individuals with and without type 2 diabetes mellitus (T2D). Beat-to-beat arterial pressure and continuous electrocardiogram were measured after a 12-hour overnight fast and in response to glucose ingestion (75 g dextrose) in obese subjects with (T2D group, n = 23) and without (OB group, n = 36) T2D before and after 16 weeks of aerobic exercise training at moderate intensity. Autonomic modulation was assessed using spectral analysis of systolic blood pressure variability (BPV), heart rate variability (HRV), and analysis of baroreflex sensitivity (BRS). Glucose ingestion significantly increased low-frequency (LF(SBP)), low-frequency HRV (LF(RRI)), and the ratio of low- to high-frequency components of HRV (LF(RRI)/HF(RRI)), and decreased the high-frequency power (HF(RRI)) (P < .05). Exercise training increased LF(RRI) and LF(RRI)/HF(RRI) responses, and reduced HF(RRI) and LF(SBP) to glucose ingestion in both groups (P < .05), but increased fasted BRS in the OB group only (P < .05); glucose intake had no effect on BRS (P > .05). In conclusion, a 16-week exercise training program improved cardiac autonomic modulation in response to an oral glucose load in obese adults, independently of diabetes status, and in the absence of remarkable changes in body weight, body composition, fitness level, and glycemic control. PMID- 20015526 TI - Outcome of repeat hepatectomy in patients with hepatocellular carcinoma aged 75 years and older. AB - BACKGROUND: We sought to evaluate the influence of age on the outcome of repeat hepatectomies in patients > or = 75 years with recurrent hepatocellular carcinoma (HCC). METHODS: We studied 121 curative repeat hepatectomies retrospectively. Among the 121 patients, 100, 20, and 1 received second, third, and fourth hepatectomies, respectively. The short-term surgical results of a younger group (<75 years; n = 88) and those of an elderly group (> or = 75 years; n = 33) were compared. The long-term prognosis of the patients who underwent second hepatectomies was also compared between a younger group (<75 years; n = 77) and an elderly group (> or = 75 years; n = 23). RESULTS: The patients in the elderly group displayed more comorbid conditions pre-operatively, including hypertension and cardiovascular diseases, than the younger group (P < .05); however, there was no significant difference in the incidence of postoperative complications or the duration of postoperative hospital stay. The long-term prognosis in the elderly group was almost identical to that in the younger group. The 3-year overall survival rates for the younger group and the elderly group were 83 vs 73% (P = .51). Disease-free, 3-year survival rates for the younger group and the elderly group were 35% vs 38% (P = .88). CONCLUSION: Our findings suggest that advanced age by itself does not have an adverse effect on operative outcomes, including postoperative complications and long-term prognosis. Repeat hepatectomy may, therefore, be justified for recurrent HCC in selected elderly patients. PMID- 20015527 TI - Unknown etiology aortic aneurysm complicated with multiple vertebral erosions and aortoenteric fistula. PMID- 20015528 TI - Effect of heat-shock protein-90 (HSP90) inhibition on human hepatocytes and on liver regeneration in experimental models. AB - BACKGROUND: Targeting heat shock protein 90 (HSP90) has gained great interest for cancer therapy. However, in view of novel multimodality therapy approaches for treating hepatic metastases, concerns have raised regarding the impact of targeted therapies on liver regeneration and repair. In this study, we investigated the impact of HSP90 inhibition on liver regeneration in murine models. METHODS: Effects of HSP90 inhibition on the activation of signaling intermediates, expression of vascular endothelial growth factor (VEGF), and hepatocyte growth factor (HGF) were investigated in primary human hepatocytes (PHHs) in vitro. Effects of HSP90 inhibition on liver regeneration and repair were determined in a murine hepatectomy model and in a model with acute carbon tetrachloride (CCl(4))-induced liver damage. RESULTS: Inhibition of HSP90 effectively diminished the constitutive phosphorylation of Akt, Erk, and STAT3 in PHHs. Conversely, inhibition of HSP90 significantly increased the expression of both VEGF and HGF mRNA, and induced HSP70 protein in PHH cultures in vitro. In vivo, HSP90 inhibition significantly upregulated constitutive VEGF mRNA and HSP70 in murine livers and did not impair liver re-growth after 70% hepatectomy. Furthermore, BrdUrd-staining and histological quantification of necrotic areas revealed that HSP90 inhibition did not impair liver regeneration following partial hepatectomy, or liver repair that occurs after toxic liver injury with CCl(4). CONCLUSION: Targeting HSP90 does not negatively affect the multifactorial process of liver regeneration and repair in vivo. Hence, the use of inhibitors to HSP90 appears to be a valid option for neoadjuvant therapy of liver metastases when subsequent surgery is intended. PMID- 20015529 TI - Methodologies for establishing validity in surgical simulation studies. AB - BACKGROUND: Validating assessment tools in surgical simulation training is critical to objectively measuring skills. Most reviews do not elicit methodologies for conducting rigorous validation studies. Our study reports current methodological approaches and proposes benchmark criteria for establishing validity in surgical simulation studies. METHODS: We conducted a systematic review of studies establishing validity. A PubMed search was performed with the following keywords: "validity/validation," "simulation," "surgery," and "technical skills." Descriptors were tabulated for 29 methodological variables by 2 reviewers. RESULTS: A total of 83 studies were included in the review. Of these studies, 60% targeted construct, 24% targeted concurrent, and 5% looked at predictive validity. Less than half (45%) of all the studies reported reliability data. Most studies (82%) were conducted in a single institution with a mean of 37 subjects recruited. Only half of the studies provided rationale for task selection. Data sources included simulator-generated measures (34%), performance assessment by human evaluators (33%), motion tracking (6%), and combined modes (28%). In studies using human evaluators, videotaping was a common (48%) blinding technique; however, 34% of the studies did not blind evaluators. Commonly reported outcomes included task time (86%), economy of motion (51%), technical errors (48%), and number of movements (25%). CONCLUSION: The typical validation study comes from a single institution with a small sample size, lacks clear justification for task selection, omits reliability reporting, and poses potential bias in study design. The lack of standardized validation methodologies creates challenges for training centers that survey the literature to determine the appropriate method for their local settings. PMID- 20015530 TI - Oxidative transformation of micropollutants during municipal wastewater treatment: comparison of kinetic aspects of selective (chlorine, chlorine dioxide, ferrate VI, and ozone) and non-selective oxidants (hydroxyl radical). AB - Chemical oxidation processes have been widely applied to water treatment and may serve as a tool to minimize the release of micropollutants (e.g. pharmaceuticals and endocrine disruptors) from municipal wastewater effluents into the aquatic environment. The potential of several oxidants for the transformation of selected micropollutants such as atenolol, carbamazepine, 17 alpha-ethinylestradiol (EE2), ibuprofen, and sulfamethoxazole was assessed and compared. The oxidants include chlorine, chlorine dioxide, ferrate(VI), and ozone as selective oxidants versus hydroxyl radicals as non-selective oxidant. Second-order rate constants (k) for the reaction of each oxidant show that the selective oxidants react only with some electron-rich organic moieties (ERMs), such as phenols, anilines, olefins, and deprotonated-amines. In contrast, hydroxyl radicals show a nearly diffusion controlled reactivity with almost all organic moieties (k>or=10(9)M(-1) s(-1)). Due to a competition for oxidants between a target micropollutant and wastewater matrix (i.e. effluent organic matter, EfOM), a higher reaction rate with a target micropollutant does not necessarily translate into more efficient transformation. For example, transformation efficiencies of EE2, a phenolic micropollutant, in a selected wastewater effluent at pH 8 varied only within a factor of 7 among the selective oxidants, even though the corresponding k for the reaction of each selective oxidant with EE2 varied over four orders of magnitude. In addition, for the selective oxidants, the competition disappears rapidly after the ERMs present in EfOM are consumed. In contrast, for hydroxyl radicals, the competition remains practically the same during the entire oxidation. Therefore, for a given oxidant dose, the selective oxidants were more efficient than hydroxyl radicals for transforming ERMs-containing micropollutants, while hydroxyl radicals are capable of transforming micropollutants even without ERMs. Besides EfOM, ammonia, nitrite, and bromide were found to affect the micropollutant transformation efficiency during chlorine or ozone treatment. PMID- 20015531 TI - Extracellular polymeric substances (EPS) in upflow anaerobic sludge blanket (UASB) reactors operated under high salinity conditions. AB - Considering the importance of stable and well-functioning granular sludge in anaerobic high-rate reactors, a series of experiments were conducted to determine the production and composition of EPS in high sodium concentration wastewaters pertaining to anaerobic granule properties. The UASB reactors were fed with either fully acidified substrate (FAS) consisting of an acetate medium (reactor R1) or partly acidified substrate (PAS) consisting of acetate, gelatine and starch medium (reactors R2, R3, and R4). For EPS extraction, the cation exchange resin (CER) method was used. Strength and particle size distribution were determined by assessing the formation of fines sludge under conditions of high shear rate and by laser diffraction, respectively. Batch tests were performed in 0.25L bottles to study Ca(2+) leaching from anaerobic granular sludge when incubated in 20g Na(+)/L in the absence of feeding for 30 days. Results show a steady increase in the bulk liquid Ca(2+) concentration during the incubation period. UASB reactor results show that the amounts of extracted proteins were higher from reactors R2 and R3, fed with PAS compared to the sludge samples from reactor R1, fed with FAS. Strikingly, the amount of extracted proteins also increased for all reactor sludges, irrespective of the Na(+) concentration applied in the feed, i.e. 10 or 20gNa(+)/L. PAS grown granular sludges showed an important increase in particle size during the operation of the UASB reactors. Results also show that, addition of 1gCa(2+)/L to the high salinity wastewater increases the granules' strength. PMID- 20015532 TI - L-glyceraldehyde 3-phosphate reductase from Escherichia coli is a heme binding protein. AB - Recently, we reported that YghZ from Escherichia coli functions as an efficient L glyceraldehyde 3-phosphate reductase (Gpr). Here we show that Gpr co-purifies with a b-type heme cofactor. Gpr associates with heme in a 1:1 stoichiometry to form a complex that is characterized by a K(d) value of 5.8+/-0.2 microM in the absence of NADPH and a K(d) value of 11+/-1.3 microM in the presence of saturating NADPH. The absorbance spectrum of reconstituted Gpr indicates that heme is bound in a hexacoordinate low-spin state under both oxidizing and reducing conditions. The physiological function of heme association with Gpr is unclear, as the L-glyceraldehyde 3-phosphate reductase activity of Gpr does not require the presence of the cofactor. Bioinformatics analysis reveals that Gpr clusters with a family of putative monooxygenases in several organisms, suggesting that Gpr may act as a heme-dependent monooxygenase. The discovery that Gpr associates with heme is interesting because Gpr shares 35% amino acid identity with the mammalian voltage-gated K+ channel beta-subunit, an NADPH dependent oxidoreductase that endows certain voltage-gated K+ channels with hemoprotein-like, O2-sensing properties. To date the molecular origin of O2 sensing by voltage-gated K+ channels is unknown and the results presented herein suggest a role for heme in this process. PMID- 20015533 TI - Dicarboxylic acid transport through hematite-coated sand. AB - To better understand the behavior of low molecular weight organic acids in subsurface environments, the transport of three dicarboxylic acids (phthalic, maleic, and fumaric acid) in water-saturated columns packed with a hematite coated sand was investigated in single and binary organic acid systems. Experiments were conducted at a single ionic strength (0.1M) and at two pH values (4.1 and 5.3). In single-acid systems, the order of breakthrough at both pH values was fumaric acid, followed by maleic acid, and then phthalic acid. The shape of the breakthrough curves for the acids at the two pH values were similar except at pH 5.3 phthalic acid showed two adsorption fronts. The initial front only partially broke through, whereas the second front proceeded to complete breakthrough. This behavior resulted from a marked pH increase during phthalic acid adsorption and suggests that the single-acid systems behaved as dual component systems with the organic acid and hydrogen ion as variables. The breakthrough curves for the binary organic acid systems showed organic acids with a higher adsorption affinity (e.g., phthalic acid) competitively displace organic acids with a lower adsorption affinity (e.g., fumaric acid). The dual-component effect observed for phthalic acid in the single-acid systems was suppressed in the mixed acid systems, perhaps reflecting pH changes that accompanied the desorption of the weakly-binding acids. These results may provide an important step toward further elucidating the processes controlling organic acid fractionation in the subsurface. PMID- 20015534 TI - Distribution of perfluoroalkyl compounds in seawater from northern Europe, Atlantic Ocean, and Southern Ocean. AB - The global distribution of perfluoroalkyl compounds (PFCs) were investigated in surface water samples collected onboard the Polarstern in Northern Europe, Atlantic and Southern Ocean (52 degrees N-69 degrees S) in 2008. The water samples were solid-phase extracted with Oasis WAX cartridges and analysed using the high-performance liquid chromatography interfaced to tandem mass spectrometry. Concentrations of various PFCs, including C(4), C(6), C(8) perfluoroalkyl sulfonates (PFSAs), perfluorooctane sulfinate (PFOSi), C(5)-C(12) perfluoroalkyl carboxylic acids (PFCA) and perfluorooctane sulfonamide (FOSA) were quantified. Perfluorooctane sulfonate (PFOS) and perfluorooctanoic acid (PFOA) were the predominant compounds with a maximum concentration of 232 and 223pgL(-1), respectively. Results indicate that industrial areas like the European Continent act as source of PFCs, while ocean water is an important as a sink as well as the transport medium of these compounds. Interestingly, in the equator area the summation operatorPFC concentration increased, which indicates that there exists an atmospheric or other unknown input source of PFCs. In the Southern Ocean only PFOS was detected which could be caused by atmospheric transport of its precursors. PMID- 20015535 TI - Micropore surface area of alkali-soluble plant macromolecules (humic acids) drives their decomposition rates in soil. AB - Previous studies suggested that micropore surface area (MSA) of alkali-soluble bio-macromolecules of aerial plant residues of maize constitutes an important factor that explains their humification in soil, that is, preservation against biological degradation. On the other hand, root plant residue contributes to the soil humus balance, as well. Following the experimental design used in a previous paper published in this journal, this study shows that the biochemical recalcitrance of the alkali-soluble acid-insoluble fraction of the root plant material, contributed to the root maize humification of both Wild-type maize plants and its corresponding mutant brown midrib (bm3), this latter characterized by reduced lignin content. Humic acids (HAs) existed in root (root-HAs) were less degraded in soil than corresponding HAs existed in shoot (shoot-HAs): shoot-HAs bm3 (48%)>shoot-HAs Wild-type (37%)>root-HAs Wild-type (33%)>root-HAs bm3 (22%) (degradability shown in parenthesis). These differences were related to the MSA of HAs, that is, root-HAs having a higher MSA than shoot-HAs: shoot-HAs bm3 (41.43+/-1.2m(2)g(-1))484 days, CI>534 days, and CI>584 days were built to explore factors of long CIs. These cut-offs were selected to represent calving-to-conception intervals of >200 days, >250 days, and >300 days. BLV-seropositive cows had higher odds of having a CI>484 days compared to BLV-seronegative cows, and BLV serostatus interacted with lactation number in this model, with 1st lactation seropositive cows being more likely to have a CI>484 days than older seropositive cows. NC seropositive cows had a 1.27 times higher odds of exhibiting a CI>484 days, a 1.37 times higher odds of a CI>534 days, and a 1.54 times higher odds of a CI>584 days, compared to NC-seronegative cows. Neither BVDV nor MAP seropositivity showed any significant effect in these models. For the FSC models, a first service was classified successful (pregnancy=1) if it was the cow's last service and she calved 270-290 days later. A mixed logistic regression model of FSC revealed an interaction between NC and BVDV-seropositivity at the herd level, with odds ratios of 0.64, 1.06 and 0.85 for NC-seropositive cows (compared to NC seronegative cows) in BVDV-seronegative, BVDV-seropositive and BVDV-missing herds, respectively. BLV and MAP seropositivity had no significant impact on FSC. All models controlled for herd-clustering effects, and included parity, linear score of somatic cell counts, peak milk, and province to control for confounding. The overall FSC was 51%, the average CI was 393 days, and 18%, 9% and 5% of lactations had CI>484 days, >534 days, and >584 days, respectively. PMID- 20015557 TI - Lack of evidence for fecal shedding of Mycobacterium avium subsp. paratuberculosis in calves born to fecal culture positive dams. AB - The objective was to detect presence of calves excreting Mycobacterium avium subsp. paratuberculosis (MAP) in their feces as a consequence of being born to MAP fecal culture positive (vs. negative) dams. For each cow that was about to calf, approximately 10g of feces was collected manually by the herdsmen from the rectum using a disposable plastic examination sleeve within 48-72h prior to actual calving. Between 1 and 3d of birth, herd personnel collected approximately 10g of fecal samples followed by monthly visits to the farm at which time 10g of fecal samples were again collected by study investigators from each calf at approximately 30, 60 and 90d of age. Mycobacterium avium subsp. paratuberculosis was recovered from 8% (5/60) of the cows that gave birth to calves. However, MAP was not recovered from any of the fecal samples (0/240) collected from study calves. Findings of the present study suggest lack of evidence for fecal excretion of MAP in calves born to fecal culture positive (vs. negative) dams in a heavily infected herd. PMID- 20015558 TI - A description of the management of itinerant grazing ducks in the Mekong river delta of Vietnam. AB - A cross-sectional survey of management practices of itinerant grazing ducks (known as field running ducks) was conducted during March 2008 in four districts of Can Tho and Bac Lieu provinces in the Mekong River Delta, Vietnam. The objective was to characterize the demographic structure of the itinerant grazing duck population and the management practices which might be related to the continuing outbreaks of H5N1 avian influenza in this region. Broiler duck flocks were owned by 55% of survey respondents, and layer flocks by 53%. Greater than 80% of ducks within 93% of villages and 99% of duck flocks were reported to have been vaccinated against H5N1 avian influenza, whereas only 19% of villages and 60% of chicken flocks had greater than 80% of chickens vaccinated. Fifty-nine percent of duck owners moved their ducks outside their home communes, whilst 37% and 28% of owners moved their ducks outside their home district and province, respectively. Larger flocks were more likely to be run outside their home district compared with smaller flocks. After adjusting for the effect of flock production type and district, the odds of an out-of-district field running duck flock movement was increased by a factor of 7.24 (95% CI 2.89-19.24) for households with flocks of more than 800 ducks, compared with flocks of less than 250. Most households sold ducks to traders (72%) or to neighbours (33%), whereas less than 20% sold their birds through markets. The findings of this study suggest that surveillance strategies for field running duck flocks should focus on layer flocks as well as larger flocks as they are more likely to be moved outside of their home district, facilitating long-distance disease spread. PMID- 20015559 TI - Monopoly pricing of an antibiotic subject to bacterial resistance. AB - We develop a dynamic bio-economic model of bacterial resistance and disease transmission in which we characterize the pricing policy of a monopolist who is protected by a patent. After expiration, the monopolist behaves competitively in a generic industry having open access to the common pool of antibiotic efficacy and infection. The monopolist manages endogenously the levels of antibiotic efficacy as well as the infected population, which represent quality and market size respectively and achieves, at least temporarily, higher such levels than a hypothetically myopic monopolist who does not take into account the dynamic externalities. The pricing policy and the biological system is characterized by the turnpike property. Before the patent vanishes, the monopolist behaves more and more myopically, leading to a continuous decrease in the price of the antibiotic. Once the generic industry takes over, a discontinuous fall in price occurs. Whether a prolongation of the patent is socially desirable depends on the relative levels of antibiotic efficacy and infection. PMID- 20015560 TI - A flexible two-part random effects model for correlated medical costs. AB - In this paper, we propose a flexible "two-part" random effects model (Olsen and Schafer, 2001; Tooze et al., 2002) for correlated medical cost data. Typically, medical cost data are right-skewed, involve a substantial proportion of zero values, and may exhibit heteroscedasticity. In many cases, such data are also obtained in hierarchical form, e.g., on patients served by the same physician. The proposed model specification therefore consists of two generalized linear mixed models (GLMM), linked together by correlated random effects. Respectively, and conditionally on the random effects and covariates, we model the odds of cost being positive (Part I) using a GLMM with a logistic link and the mean cost (Part II) given that costs were actually incurred using a generalized gamma regression model with random effects and a scale parameter that is allowed to depend on covariates (cf., Manning et al., 2005). The class of generalized gamma distributions is very flexible and includes the lognormal, gamma, inverse gamma and Weibull distributions as special cases. We demonstrate how to carry out estimation using the Gaussian quadrature techniques conveniently implemented in SAS Proc NLMIXED. The proposed model is used to analyze pharmacy cost data on 56,245 adult patients clustered within 239 physicians in a mid-western U.S. managed care organization. PMID- 20015561 TI - Zinc finger nuclease technology heralds a new era in mammalian transgenesis. AB - Non-mouse mammalian transgenesis is limited by its overall inefficiency and technical hurdles. Recent years have seen the emergence of two approaches that are applicable to most mammals. The first, based on lentivirus vectors, allows efficient generation of transgenic founders, most of them expressing the transgene. The second, recently applied to produce transgenic fish and mammals, takes advantage of the design of specific 'DNA-scissors' for efficient introduction of subtle mutations in potentially any region of the genome. This review focuses on the potential of this latter technology to modify mammalian genomes without the need to apply challenging and less-efficient protocols. We highlight the complementary aims of these new approaches and the as-yet unexplored possibilities offered by their combination. PMID- 20015562 TI - Influence of the hot-fill water-spray-cooling process after continuous pasteurization on the number of decimal reductions and on Alicyclobacillus acidoterrestris CRA 7152 growth in orange juice stored at 35 degrees C. AB - In this study, the influence of the hot-fill water-spray-cooling process after continuous pasteurization on the number of decimal reductions (gamma) and growth parameters (lag time; lambda, ratio N(f)/N(o); kappa, maximum growth rate; mu) of Alicyclobacillus acidoterrestris CRA 7152 in orange juice stored at 35 degrees C were investigated. Two different inoculum levels of A. acidoterrestris CRA 7152 (10(2) and 10(3) spores/mL) in orange juice (11(0)Brix, pH 3.7) and a Microthermics UHT-HTST pilot plant were used to simulate industrial conditions. Results have shown that regardless of the inoculum level (10(2) or 10(3) spores/mL), the pasteurization processes were unable to cause even 1 gamma. Predictive modeling using the Baranyi model showed that only kappa and time to reach 10(4)spores/mL (t10(4) - time to juice spoilage) were affected by the spore inoculum used (p<0.05). It has been concluded that A. acidoterrestris was able to survive the hot-fill process and to grow and spoil orange juice in 5-6 days when the final storage temperature was 35 degrees C. PMID- 20015563 TI - Influence of Origanum vulgare L. essential oil on enterotoxin production, membrane permeability and surface characteristics of Staphylococcus aureus. AB - This study evaluated the influence of the essential oil from Origanum vulgare L. on the enterotoxin production, membrane permeability and cell surface characteristics of Staphylococcus aureus. The suppression of enterotoxin production occurred totally in the broth added with the essential oil at subinhibitory concentrations (0.3 and 0.15 microL/mL). Loss of 260-nm-absorbing material and potassium ions occurred immediately after addition of the essential oil at 0.6 and 1.2 microL/mL and followed up to 120 min. Electron microscopy of essential oil-treated cells revealed the formation of roles in the cell surfaces and loss of cytoplasm material. According to these results, O. vulgare essential oil could be rationally applied in food products both to inhibit the growth of S. aureus and to suppress the synthesis of staphylococcal enterotoxins. PMID- 20015564 TI - A novel glucokinase gene mutation and its effect on glycemic/C-peptide fluctuations in a patient with maturity-onset diabetes of the young type 2. AB - Maturity-onset diabetes of the young (MODY) is a group of disorders accounting for 2-5% of diabetes; MODY2 is caused by inactivating GCK mutations. We report a case of MODY2 caused by a novel GCK mutation and demonstrate differential glycemic/C-peptide responses to treatment with insulin, no medication, and an oral sulfonylurea. PMID- 20015565 TI - The expression level of non-alcoholic fatty liver disease-related gene PNPLA3 in hepatocytes is highly influenced by hepatic lipid status. AB - BACKGROUND & AIMS: Recent studies have suggested that variations in PNPLA3 are associated with non-alcoholic fatty liver disease (NAFLD). To gain insight into the potential function of PNPLA3 in liver, we have determined the effect of metabolic shifts on the hepatic expression profile of PNPLA3 in mice. METHODS: PNPLA3 expression in wild-type C57BL/6 and NAFLD-susceptible LDL receptor knockout (LDLR-/-) mice was determined using microarray and real-time PCR analysis. RESULTS: PNPLA3 expression in livers is 50- to 100-fold lower as compared to (cardiac) muscle and adipose tissue in regular chow diet-fed mice. Feeding a Western-type diet stimulated hepatic relative PNPLA3 expression level 23-fold (p<0.001) both in C57BL/6 mice and LDLR-/- mice, suggesting that PNPLA3 does become an important player in hepatic lipid metabolism under conditions of lipid excess. Subjecting mice to fasting fully reversed the effect of the Western type diet on hepatic PNPLA3 expression. Under these conditions, the expression level of PNPLA3 in adipose tissue is also decreased 90% (p<0.001). Cellular distribution analysis revealed that PNPLA3 is expressed in hepatocytes but not in liver endothelial and Kupffer cells. Microarray-based gene profiling showed that the expression level of PNPLA3 in hepatocytes is correlated with that of genes associated with the lipogenic pathway such as ME1, SPOT14, and SCD1. CONCLUSIONS: It appears that the NAFLD-related gene PNPLA3 is highly responsive to metabolic changes in hepatocytes within the liver and its relative change in expression level suggests an essential function in lipogenesis. PMID- 20015567 TI - Hepatitis C virions subvert natural killer cell activation to generate a cytokine environment permissive for infection. AB - BACKGROUND & AIMS: Hepatitis C virus (HCV) is remarkably successful in establishing persistent infections due to its ability to evade host immune responses through a combination of mechanisms including modulation of interferon (IFN) signalling in infected cells, interference with effector cell function of the immune system and continual viral genetic variation. We have previously demonstrated that natural killer (NK) cells can be inhibited in vitro by recombinant HCV glycoprotein E2 via cross-linking of CD81, a cellular co-receptor for the virus. METHODS: Taking advantage of the recently established tissue culture system for HCV, we have studied the effects of CD81 engagement by the HCV envelope glycoprotein E2 when the protein is part of complete, infectious viral particles. Specifically, we asked whether exposure to HCV viral particles (HCVcc) affects activation of NK cells and whether altered NK cell activation, in turn, impacts on HCV infectivity. RESULTS: We found that immobilized HCVcc, unlike soluble HCVcc, inhibited IFN-gamma production by interleukin (IL)-12 activated NK cells, and that this effect was mediated by engagement of cellular CD81 by HCV virion displayed E2. In contrast, NK-production of IL-8 was increased in the presence of HCV. The cytokines produced by IL-12 activated NK cells strongly reduced the establishment of productive HCV infection. Importantly, NK-cell derived cytokines secreted in the presence of HCVcc showed a diminished antiviral effect that correlated with IFN-gamma reduction, while IL-8 concentrations had no impact on HCV infectivity. CONCLUSIONS: Exposure to HCVcc modulates the pattern of cytokines produced by NK cells, leading to reduced antiviral activity. PMID- 20015568 TI - Disinvestment for re-allocation: a process to identify priorities in healthcare. AB - Resource scarcity and increasing service demand lead health systems to cope with choices within constrained budgets. The aim of the paper is to describe the study carried out in the Tuscan Health System in Italy on how to set priorities in the disinvestment process for re-allocation. The analysis was based on 2007 data benchmarking of the Tuscan Health System with an impact on the level of resources used. For each indicator, the first step was to estimate the gap between the performance of each Health Authority (HA) and the best performance or the regional average. The second step was to measure this gap in terms of financial value. The results of the analysis demonstrated that, at the regional level, 2-7% of the healthcare budget can be re-allocated if all the institutions achieve the regional average or the best practice. The implications of this study can be useful for policy makers and the HA top management. In the context of resource scarcity, it allows managers to identify the areas where the institutions can achieve a higher level of efficiency without negative effects on quality of care and instead re-allocate resources toward services with more value for patients. PMID- 20015569 TI - An approach to health system strengthening in the Union of Myanmar. AB - OBJECTIVES: In 2007 and 2008, Myanmar developed a health system strengthening (HSS) strategy and proposal through funding support from the Global Alliance for Vaccines and Immunization (GAVI). The aim of this paper is to identify critical success factors in the development of the HSS strategy in Myanmar. METHODS: The main source of information for this review includes international and national literature, and participant observation by the authors in the health systems analysis and HSS strategy development in Myanmar between 2007 and 2009. RESULTS: Critical success factors in the development of the HSS strategy included evidence based development of the strategy through a sector analysis, and a long-term approach to strategy development with wide stakeholder participation. This contributed to important strategy breakthroughs in the areas of health planning, health financing, human resource management and civil society partnerships. CONCLUSION: Implementation of the HSS strategy in Myanmar should position the MOH and partners well to implement challenging system reforms in the areas of health planning, financing and human resource management in the coming years, as well as support more coordinated efforts for relief and recovery effort following the Nargis natural disaster in 2008. These innovations in Myanmar, with evidence of similar breakthroughs in other countries of the Asian region including North Korea, Cambodia, Nepal and Sri Lanka, provides promising evidence of the potential of the HSS approach as an emerging health development paradigm, particularly in relation to responding to the issue of "within country" inequities in access to health care. PMID- 20015570 TI - Assessing erectile neurogenic dysfunction from heart rate variability through a Generalized Linear Mixed Model framework. AB - BACKGROUND: The low (LF) vs. high (HF) frequency energy ratio, computed from the spectral decomposition of heart beat intervals, has become a major tool in cardiac autonomic system control and sympatho-vagal balance studies. The (statistical) distributions of response variables designed from ratios of two quantities, such as the LF/HF ratio, are likely to non-normal, hence preventing e.g., from a relevant use of the t-test. Even using a non-parametric formulation, the solution may be not appropriate as the test statistics do not account for correlation and heteroskedasticity, such as those that can be observed when several measures are taken from the same patient. OBJECTIVES: The analyses for such type of data require the application of statistical models which do not assume a priori independence. In this spirit, the present contribution proposes the use of the Generalized Linear Mixed Models (GLMMs) framework to assess differences between groups of measures performed over classes of patients. METHODS: Statistical linear mixed models allow the inclusion of at least one random effect, besides the error term, which induces correlation between observations from the same subject. Moreover, by using GLMM, practitioners could assume any probability distribution, within the exponential family, for the data, and naturally model heteroskedasticity. Here, the sympatho-vagal balance expressed as LF/HF ratio of patients suffering neurogenic erectile dysfunction under three different body positions was analyzed in a case-control protocol by means of a GLMM under gamma and Gaussian distributed responses assumptions. RESULTS: The gamma GLMM model was compared with the normal linear mixed model (LMM) approach conducted using raw and log transformed data. Both raw GLMM gamma and log transformed LMM allow better inference for factor effects, including correlations between observations from the same patient under different body position compared to the raw LMM. The gamma GLMM provides a more natural distribution assumption of a response expressed as a ratio. CONCLUSIONS: A gamma distribution assumption intrinsically models quadratic relationships between the expected value and the variance of the data avoiding prior data transformation. SAS and R source code are available on request. PMID- 20015571 TI - Effect of a virtual reality interface on the learning curve and on the accuracy of a surgical planner for total hip replacement. AB - The aim of this study is to evaluate the performance of a non-conventional input and output device (virtual reality) in a total hip replacement surgical planner. A test was performed asking five users to position a cup in a defined position. Every user performed the task using three different hardware configurations: (I) conventional mouse and monitor, (II) mouse and auto-stereoscopic monitor, and (III) 12-DOF tracker (haptic device) and auto-stereoscopic monitor. The results were evaluated in terms of root mean square error of the obtained position with respect to the target one and in terms of learning curve. The results showed that the examined VR technology does not show a sufficient positioning accuracy to be considered for clinical assessment. PMID- 20015572 TI - Oral Chinese herbal medicine (CHM) as an adjuvant treatment during chemotherapy for non-small cell lung cancer: A systematic review. AB - BACKGROUND: Non-small cell lung cancer (NSCLC) remains a major global health problem because of its prevalence and poor prognosis. Treatment options are limited and there is a need to explore alternatives. This systematic review evaluates the role of Chinese herbal medicine (CHM) in association with chemotherapy for NSCLC. METHODS: English and Chinese databases were searched for RCTs comparing CHM with conventional biomedical treatment or placebo. Papers were reviewed systematically and data were analysed using standard Cochrane software Revman 5. RESULTS: Fifteen Chinese trials involving 862 participants met the inclusion criteria. All trials were of poor quality with a considerable risk of bias. There was a significant improvement in quality of life (QoL) (increased Karnofsky Performance Status) (RR 1.83, 95% CI 1.41-2.38, p<0.00001 for both stages III, IV only NSCLC and all stages NSCLC) and less anaemia (RR 0.37, 95% CI 0.15-0.91, p=0.03 for stages III, IV only NSCLC; p=0.005 for all stages NSCLC) and neutropenia (RR 0.42, 95% CI 0.22-0.82, p=0.01 for stages III, IV only NSCLC; p<0.00001 for all stages NSCLC) when CHM is combined with chemotherapy compared to chemotherapy alone. There was no significant difference in short term effectiveness and limited inconclusive data concerning long term survival. Five promising herbs have been identified. CONCLUSION: It is possible that oral CHM used in conjunction with chemotherapy may improve QoL in NSCLC. This needs to be examined further with more rigorous methodology. PMID- 20015573 TI - A biogeochemical model of contaminant fate and transport in river waters and sediments. AB - A quasi-two-dimensional model is presented for simulating transport and transformation of contaminant species in river waters and sediments, taking into account the effect of both biotic and abiotic geochemical reactions on the contaminant fate and mobility. The model considers the downstream transport of dissolved and sediment-associated species, and the mass transfer with bed sediments due to erosion and resuspension, using linked advection-dispersion reaction equations. The model also couples both equations to the reactive transport within bed sediment phases. This is done by the use of a set of vertical one-dimensional columns representing sediment layers that take into account the reactive transport of chemicals, burial, sorption/desorption to/from the solid phase, and the diffusive transport of aqueous species. Kinetically controlled reversible solid-water mass exchange models are adopted to simulate interactions between suspended sediments and bulk water, as well as the mass exchange between bed sediments and pore water. An innovative multi-time step approach is used to model the fully kinetic nonlinear reaction terms using a non iterative explicit method. This approach enables the model to handle fast and near-equilibrium reactions without a significant increase in computational burden. At the end, two demonstration cases are simulated using the model, including transport of a sorbing, non-reactive trace metal and nitrogen cycling, both in the Colusa Basin Drain in the Central Valley of California. PMID- 20015574 TI - Experimental and modeling investigation of multicomponent reactive transport in porous media. AB - We present an experimental and modeling study of solute transport in porous media in the presence of mixing-induced precipitation of a solid phase. Conservative and reactive transport experiments were performed in a quasi-two-dimensional laboratory flow cell, filled with homogeneous and heterogeneous porous media. Conservative experiments were performed by injecting solutions containing sodium chloride and calcium chloride into the domain. In reactive transport experiments, inlet solutions of calcium chloride and sodium carbonate were injected in parallel, resulting in calcium carbonate precipitation where the solutions mix. Experimental results were used as a benchmark to examine the performance of a reactive transport numerical model. Good agreement between model predictions and experimental results was obtained for the conservative transport experiments. The reactive transport experiments featured the formation of a calcium carbonate mineral phase within the mixing zone between the two solutions, which controlled the spatial evolution of calcium carbonate in the domain. Numerical simulations performed on high resolution grids for both the homogeneous and heterogeneous porous systems underestimated clogging of the system. Although qualitative agreement between model results and experimental observations was obtained, accurate model predictions of the spatial evolution of calcium concentrations at sample points within the flow cell could not be achieved. PMID- 20015575 TI - Fractal analysis of amyloid plaques in Alzheimer's disease patients and mouse models. AB - The varied morphological and biochemical forms in which amyloid deposits in brain of Alzheimer's disease (AD) patients are complex and their mechanisms of formation are not completely understood. Here we investigated the ability of fractal dimension (FD) to differentiate between the textures of commonly observed amyloid plaques in sporadic and familial AD patients and aged-control individuals as well as in transgenic mouse models of amyloidosis. Studying more than 6000 amyloid plaques immunostained for total Abeta (Abetat), Abeta40 or Abeta42, we show here that Abeta40 FD could efficiently differentiate between (i) AD patients and aged-control individuals (P<0.001); (ii) sporadic and familial AD due to presenilin-1 or APP (A692G) mutations (P<0.001); and (iii) three transgenic mouse models of different genotypes (P<0.001). Furthermore, while diffuse and dense core plaques present in humans and transgenic mice had comparable FDs, both Abetat and Abeta42 FD could also differentiate diffuse plaques from other plaque types in both species (P<0.001). Our data suggest that plaque FD could be a valuable tool for objective, computer-oriented AD diagnosis as well as for genotype-phenotype correlations of AD. PMID- 20015576 TI - Differential recognition of vascular and parenchymal beta amyloid deposition. AB - By phage display, llama-derived heavy chain antibody fragments were selected from non-immune and immune libraries and tested for their affinity and specificity for beta amyloid by phage-ELISA, immunohistochemistry and surface plasmon resonance. We identified eight distinct heavy chain antibody fragments specific for beta amyloid. While three of them recognized vascular and parenchymal beta amyloid deposits, the remaining five heavy chain antibody fragments recognized vascular beta amyloid specifically, failing to bind to parenchymal beta amyloid. These heavy chain antibody fragments, selected from different libraries, demonstrated differential affinity for different epitopes when used for immunohistochemistry. These observations indicate that the llama heavy chain antibody fragments are the first immunologic probes with the ability to differentiate between parenchymal and vascular beta amyloid aggregates. This indicates that vascular and parenchymal beta amyloid deposits are heterogeneous in epitope presence/availability. The properties of these heavy chain antibody fragments make them potential candidates for use in in vivo differential diagnosis of Alzheimer disease and cerebral amyloid angiopathy. Continued use and characterization of these reagents will be necessary to fully understand the performance of these immunoreagents. PMID- 20015577 TI - [Optimization of sentinel lymph node biopsy in breast cancer by intraoperative axillary palpation]. AB - INTRODUCTION: Sentinel node biopsy (SNB) by radioisotopes is a widely accepted and reliable surgical method for staging breast cancer in patients with unknown positive axillary lymph nodes involvement. The main limitation of this method is due to the appearance of false negatives that may be caused by tumor lymph node blockage of the sentinel lymph node and uptake in the neighboring lymph nodes. Infiltered sentinel nodes are generally increased in size and firm. Thus, they can be detected by intraoperative palpation, even when there is no uptake by the radiotracer. AIM: To reduce the false negative rates by applying intraoperative axillary palpation after SNB. METHOD: Over a two-year period, we complemented the SNB in 168 patients with careful intraoperative axillary palpation, detecting and removing all the palpable suspicious lymph nodes (SLN) that were analyzed as sentinel nodes RESULTS: In 32 out of 168 patients, 50 palpable SLN were found. In 3 out of 32 patients, 4 infiltrated SLNs were demonstrated with negative SNB and positive axillary lymphadenectomy. Thus, intraoperative palpation avoided false negative results. In one patient, one palpable SLN with tumor involvement was observed and SNB was also positive. In the remaining 28 patients, the histological analysis of 45 SLN was negative for tumor but SNB was positive in 3 patients. CONCLUSION: Intraoperative axillary palpation, once the SNB was done, reduced the false negative rate. Thus, we consider that it should be included as one more part of this procedure. PMID- 20015578 TI - New pH-dependent complexes, from mononuclear Pd(II) monomer to heteronuclear [Pd(II),K(I)]Polymer: DNA cleavage and cytotoxicity in vitro. AB - Two novel complexes, namely, [Pd(pdc)(2)].(dmp).6H(2)O (1) and [Pd(pdc)(2)K(H(2)O)(5).3H(2)O]n (2) [H(2)pdc=pyridine-2,3-dicarboxylate acid, dmp=2,9-Dimethyl-1,10-phenanthroline] were synthesized and characterized as pH dependent products. Gel electrophoresis assay demonstrated the ability of the complexes to cleave the pBR 322 plasmid DNA. Values of IC(50) calculated for complexes 1 and 2 show that the two complexes exhibit good cytotoxic activity against different cell lines tested in general, especially more effective against HL-60 and A2780(cisR) cell lines. PMID- 20015579 TI - Educating for ethical leadership. AB - In this article we consider the nature of ethical leadership in nursing. An appreciation of the basis of such leadership requires an understanding of responsibility and of key intellectual and ethical qualities or virtues. We examine some of the educational and practice strategies to promote ethical leadership. We argue that there are different levels of ethical leadership. All members of the nursing workforce are ethical leaders in so far as they demonstrate a commitment to ethical practice in their everyday work and act as ethical role models for others. Nurse managers are responsible for influencing their team and for acting as arbiters between organisational and professional values. Nurse educators are role models and ethical leaders as they ensure that the explicit and hidden curriculum demonstrate a commitment to professional values. Nurses who assume political roles have an obligation to lead on ethical agenda compatible with the values of nursing. PMID- 20015580 TI - Update on melanoma: the present position. AB - Cutaneous melanoma is becoming increasingly common among people with fair skins, and this has been blamed on exposure to ultraviolet radiation from sunbathing, the use of sun beds, and holidays abroad. The key to controlling the epidemic of melanoma is prevention, but also important are the organisation of the skin cancer service, surgical advances, and the development of neoadjuvant treatments. Because there are many of these tumours in the head and neck region, head and neck surgeons are increasingly involved in the management of such patients. We review the current philosophy in the management of melanoma. PMID- 20015581 TI - Effects of foot orthoses on gait patterns of flat feet patients. AB - BACKGROUND: Although foot orthotics are widely prescribed for the treatment of flatfoot, the biomechanical effects of such devices are not yet fully clear. Accordingly, this study conducted an experimental investigation to evaluate the effects of orthoses on the gait patterns of patients with flatfoot during level walking. METHODS: Eleven adults with flatfoot deformities were recruited. For each participant, kinematic and kinetic data were measured under three test conditions, i.e. walking barefoot, walking with shoes, and walking with shoes and insoles. During each test, the participants' gaits patterns were recorded and analyzed using a motion analysis system, two Kistler force plates and EVaRT software. FINDINGS: The results showed that walking with shoes and insoles and walking with shoes conditions increased the peak ankle dorsiflexion angle and moment, and also reduced the peak ankle plantarflexion angle and moment. Furthermore, walking with shoes and insoles and walking with shoes conditions increased the peak knee varus moment. The effects of the orthoese on knee and hip were minimal and no significant differences were observed between walking with shoes and insoles and walking with shoes. INTERPRETATION: The results suggested that the foot insoles and shoes developed in this study might benefit the ankle joint in patients with flat feet. In view of the minimal changes between walking with shoes and insoles and walking with shoes, further studies may be required to clarify the interaction between the foot and the insole/shoe. PMID- 20015582 TI - Roll-over shapes of the ankle-foot and knee-ankle-foot systems of able-bodied children. AB - BACKGROUND: The roll-over shape is the effective rocker shape that a lower limb system conforms to during a step. The roll-over shape concept has been explored in detail in adults and has been successfully used in the design, evaluation, and alignment of lower limb prostheses and orthoses. No such analysis exists for the pediatric population. Therefore, the purpose of this study was to investigate the ankle-foot and knee-ankle-foot roll-over shapes in able-bodied children, values that could serve as tools for design and evaluation of lower limb pediatric prostheses and orthoses. METHODS: This study describes a multi-center retrospective review of existing motion analysis data (n=153 from three centers). Roll-over shapes were calculated by transforming center of pressure data from a laboratory-based coordinate system into two body-based coordinate systems. Roll over shapes were then characterized using a circular arc model. Best-fit radii of roll-over shapes for children in three age groups (3-7 years, 8-11 years, and 12 17 years) were compared using the Kruskal-Wallis test. FINDINGS: No significant changes were found in roll-over shape radii between the three age groups (P=0.54 for ankle-foot roll-over shape radii; P=0.12 for knee-ankle-foot roll-over shape radii). The weighted mean of median radii for ankle-foot and knee-ankle-foot roll over shapes from the three centers were approximately 22% and 17% of body stature, values similar to those seen in adults. INTERPRETATION: Children produce nearly circular knee-ankle-foot roll-over shapes at a young age that are similar to those seen in adults when scaled by body stature. PMID- 20015583 TI - Does nitrogen deposition increase forest production? The role of phosphorus. AB - Effects of elevated N deposition on forest aboveground biomass were evaluated using long-term data from N addition experiments and from forest observation plots in Switzerland. N addition experiments with saplings were established both on calcareous and on acidic soils, in 3 plots with Fagus sylvatica and in 4 plots with Picea abies. The treatments were conducted during 15 years and consisted of additions of dry NH4NO3 at rates of 0, 10, 20, 40, 80, and 160 kg N ha(-1) yr( 1). The same tree species were observed in permanent forest observation plots covering the time span between 1984 and 2007, at modeled N deposition rates of 12 46 kg N ha(-1) yr(-1). Experimental N addition resulted in either no change or in a decreased shoot growth and in a reduced phosphorus concentration in the foliage in all experimental plots. In the forest, a decrease of foliar P concentration was observed between 1984 and 2007, resulting in insufficient concentrations in 71% and 67% of the Fagus and Picea plots, respectively, and in an increasing N:P ratio in Fagus. Stem increment decreased during the observation period even if corrected for age. Forest observations suggest an increasing P limitation in Swiss forests especially in Fagus which is accompanied by a growth decrease whereas the N addition experiments support the hypothesis that elevated N deposition is an important cause for this development. PMID- 20015584 TI - Emotion-regulation strategies across psychopathology: A meta-analytic review. AB - We examined the relationships between six emotion-regulation strategies (acceptance, avoidance, problem solving, reappraisal, rumination, and suppression) and symptoms of four psychopathologies (anxiety, depression, eating, and substance-related disorders). We combined 241 effect sizes from 114 studies that examined the relationships between dispositional emotion regulation and psychopathology. We focused on dispositional emotion regulation in order to assess patterns of responding to emotion over time. First, we examined the relationship between each regulatory strategy and psychopathology across the four disorders. We found a large effect size for rumination, medium to large for avoidance, problem solving, and suppression, and small to medium for reappraisal and acceptance. These results are surprising, given the prominence of reappraisal and acceptance in treatment models, such as cognitive-behavioral therapy and acceptance-based treatments, respectively. Second, we examined the relationship between each regulatory strategy and each of the four psychopathology groups. We found that internalizing disorders were more consistently associated with regulatory strategies than externalizing disorders. Lastly, many of our analyses showed that whether the sample came from a clinical or normative population significantly moderated the relationships. This finding underscores the importance of adopting a multi-sample approach to the study of psychopathology. PMID- 20015585 TI - How to explain receptivity to conjunction-fallacy inhibition training: evidence from the Iowa gambling task. AB - Intuitive predictions and judgments under conditions of uncertainty are often mediated by judgment heuristics that sometimes lead to biases. Using the classical conjunction bias example, the present study examines the relationship between receptivity to metacognitive executive training and emotion-based learning ability indexed by Iowa Gambling Task (IGT) performance. After completing a computerised version of the IGT, participants were trained to avoid conjunction bias on a frequency judgment task derived from the works of Tversky and Kahneman. Pre- and post-test performances were assessed via another probability judgment task. Results clearly showed that participants who produced a biased answer despite the experimental training (individual patterns of the biased --> biased type) mainly had less emotion-based learning ability in IGT. Better emotion-based learning ability was observed in participants whose response pattern was biased --> logical. These findings argue in favour of the capacity of the human mind/brain to overcome reasoning bias when trained under executive programming conditions and as a function of emotional warning sensitivity. PMID- 20015586 TI - [Radiologic findings of pulmonary involvement of type B Niemann-Pick disease]. PMID- 20015587 TI - Efficient prehospital induction of therapeutic hypothermia via intraosseous infusion. PMID- 20015588 TI - Increase in plasma glucagon, a factor in hyperglycemia, is related to neurological outcome in postcardiac-arrest patients. AB - AIM OF THE STUDY: In postcardiac-arrest (PCA) patients, hyperglycemia is a factor reflecting an unfavorable outcome, and might be caused by the inflammation and stress of "sepsis-like" syndrome. In this study, plasma glucagon, a representative glycogenolytic and gluconeogenic hormone, was measured and assessed the correlation for neurological outcome in PCA patients. METHODS: This study was a retrospective, single-medical-center analysis, conducted in the intensive care unit of a university hospital. Twenty-four sequential PCA patients were included. Plasma samples were collected from the patients on days 1, 2, and 3 after the return of spontaneous circulation (ROSC). Glucagon was compared in patients with favorable and unfavorable neurological outcomes. RESULTS: At all time points, plasma glucagon was significantly higher in patients with an unfavorable outcome (P<0.05). Glucagon on day 1 had remarkable sensitivity (88.2%) and specificity (85.8%) as an indicator of outcome, and correlated with the collapse-ROSC interval, the start of cardiopulmonary resuscitation (CPR)-ROSC interval, and the epinephrine dose during CPR. CONCLUSIONS: Plasma glucagon reflects unfavorable outcomes in PCA patients, and might be related to ischemic and reperfusion stress. PMID- 20015589 TI - Evaluation of an extensively hydrolysed casein formula (Damira 2000) in children with allergy to cow's milk proteins. AB - OBJECTIVE: To assess the allergenic capacity of a new extensively hydrolysed casein formula (Damira 2000) in vivo in children with allergy to cow's milk, and to conduct an immunochemical evaluation of the product. PATIENTS AND METHODS: The study comprised 67 children (1 month-7 years) with allergy to cow's milk proteins (ACMP). Skin testing was made with whole milk, milk formula for infants, the study hydrolysate and the milk fractions (alpha-lactoalbumin, beta-lactoglobulin and casein). Specific IgE against these allergens, and oral provocation test were also performed. Immunochemical evaluation of the product was carried out with gel filtration chromatography, 4-15% acrylamide gradient gel electrophoresis, specific IgE quantification versus the casein hydrolysate and a study of its allergenic potency. RESULTS: The hydrolysate was tolerated by 66 of the 67 patients (98.5%) with ACMP. Biochemical analysis of the product confirmed the absence of traces of whole milk proteins. Specific IgE against the hydrolysate proved negative in all cases, and it was unable to inhibit FEIA even at concentrations 10 times greater than those used in the whole milk inhibition control. Likewise, no immunoblotting inhibition was recorded. CONCLUSIONS: The new extensively hydrolysed casein product is safe and well tolerated by most children with ACMP. However, as with other extensive hydrolysates, some highly sensitised patients may present clinical manifestations. Controlled tolerance testing is therefore advised, under specialised medical supervision. PMID- 20015590 TI - Open to debate. The motion: GnRH antagonists are the new way forward in hormonal therapy. PMID- 20015591 TI - Consequence assessment of indoor dispersion of sarin--a hypothetical scenario. AB - The objective of this study is to provide a hypothetical scenario of indoor dispersion of the highly toxic nerve agent sarin in a large building, which can be used as a starting point for discussion, planning, training and exercises for emergency services and responsible authorities. The indoor dispersion has been simulated using a Computational Fluid Dynamics (CFD) approach. Possible consequences have been calculated based on concentration and dose profiles. Mild intoxication effects appear within minutes, while serious injuries and fatalities occur approximately 20 min after release. Anticipated key emergency response challenges are: (i) the time factor due to rapid onset of symptoms, (ii) the large number of casualties, and (iii) the contaminated hazard scene. PMID- 20015592 TI - Industrial hazardous waste management in Turkey: current state of the field and primary challenges. AB - A holistic evaluation of a country's hazardous waste management (HWM) practices is useful in identifying the necessary actions to focus on. Based on an analysis of industrial hazardous waste (HW) generation in Turkey, this paper attempts to critically evaluate and report current Turkish HWM practices and discuss the primary challenges to be addressed. The generation of industrial HW for Turkey reported in 2004 was 1.195 million tons, which accounted for 7% of the total industrial solid waste (ISW) generated by the manufacturing industry, and for nearly 4.9% of the total solid waste generated in the country. The HW generated by the top five manufacturing product categories--basic metals, chemicals and chemical products, food and beverages, coke and refined petroleum, motor vehicles and trailers--accounted for 89.0% of total industrial HW. 21% of the HW generated in 2004 was recycled or reused, and 6% was sold or donated, whereas 73% was sent to ultimate disposal. 67% of the HW sent to ultimate disposal was disposed of at municipal landfills. The total capacity of the existing regional HW facilities is 212,500 tons/year, which accounts for about 24% of the HW to be disposed. Turkey has identified the HW problem in the country and enacted legislation, designated a lead agency, and promulgated rules and regulations. Several new initiatives are planned for improving HW management nationally; however, some HWM problems will be persistent due to previous and existing industrial development plans. These development policies led to the concentration of industry in regions marked by precious agricultural fields and high population density. This occurred because the government previously exhibited a default prioritization towards industrial development, leading to insufficient implementation of regulations on HW generators. Some of the problems may also be rooted in other countries that allow illegal trans boundary HW movements despite international regulations. PMID- 20015593 TI - Treatment of the pregnant mother with cancer: a systematic review on the use of cytotoxic, endocrine, targeted agents and immunotherapy during pregnancy. Part I: Solid tumors. AB - The association of cancer and pregnancy is increasingly encountered nowadays in clinical practice. Due to the relative rarity of the situation, it lacks a systematized approach. Different systemic therapies are used in managing cancer with uncertainty regarding the potential hazards they could pose on the pregnancy and/or the fetus. We have performed a systematic review of literature to identify all reports addressing cancer patients who were exposed to any of the known systemic therapies during the course of the pregnancy. The results were discussed in two parts; part I addresses pregnant patients with solid tumors while part I for those with hematological malignancies. In part I, we identified different solid tumors diagnosed and treated during the course of pregnancy. Breast cancer was the most commonly treated followed by ovarian cancer. Other tumors were treated as well including lung cancer, cervical cancer, sarcoma and melanomas. It is important to acknowledge the intent of therapy (palliative vs. curative) and the patients has to be properly counseled to reach an informed decision. We aim to provide a more robust consensus on how to approach these cases and provide a higher degree of evidence to support the safety of applying certain management strategies over the other. PMID- 20015594 TI - SRC kinase inhibition: targeting bone metastases and tumor growth in prostate and breast cancer. AB - Prostate and breast cancer cells preferentially metastasize to bone, whereupon a complex interaction between metastatic tumor cells, osteoclasts, and osteoblasts results in the development of bone lesions that cause significant pain and patient morbidity. For patients with bone lesions, the goals of treatment are to decrease tumor growth, prevent further metastases, and inhibit tumor-associated bone pathology. Preclinical data suggest that SRC, a nonreceptor tyrosine kinase, is an important signaling molecule during the processes of osteoclast-mediated bone resorption, tumor growth, and metastasis, and that SRC has a role in hormone receptor signaling and resistance. As such, SRC represents a logical target for the treatment of advanced metastatic prostate or breast cancer. SRC-targeting agents, including dasatinib, saracatinib, and bosutinib, are currently in clinical development for patients with solid tumors. Preliminary data from phase 1/2 trials, including tumor responses and bone-specific activity in patients with prostate or breast cancer, demonstrate that SRC inhibitors have potential in the clinical setting. Data arising from ongoing and future clinical trials will confirm whether SRC inhibitors provide clinical benefits for patients with advanced disease. PMID- 20015595 TI - Decrease of CD4(+)FOXP3(+) T regulatory cells in the peripheral blood of human subjects undergoing a mental stressor. AB - We have previously shown that acute psychological stress alerts the adaptive immune response causing an increase in antigen-experienced effector T cells in the peripheral blood. T regulatory cells (Tregs) play a central role in maintaining self-tolerance and controlling autoimmune responses. Here, we analyzed for the first time the behaviour of Tregs in the context of a stress induced activation of the adaptive immune response. 31 healthy young males underwent a brief laboratory stressor and, in a crossover design, served as their own unstressed controls. We quantified effects of acute stress on CD4(+)FOXP3(+) T regulatory cells and other T cell subpopulations using flow cytometry. In addition, the expression of Treg-related effector molecules and stress hormone receptors were analyzed in the subjects' peripheral T cells. We confirmed our previous observation of a stress-induced decrease in CD45RA(+)CCR7(+) "naive" and CD45RA(-)CCR7+ "central memory" T cells while CD45RA(-)-CCR7(-) "memory effector" and CD45RA(+)CCR7(-) "terminally differentiated" effector T cells remained stable or increased. Importantly, we found acute psychological stress to cause a concomitant decrease in CD4(+)FOXP3(+) Tregs and in CD4(+) T cells expressing Treg-related effector molecules cytotoxic T-lymphocyte antigen-4 (CTLA-4) and latency associated peptide (LAP). Finally, we observed beta(1)-adrenergic and glucorticoid alpha receptors to be overexpressed in Tregs, suggesting that these molecules might mediate stress-related effects on Tregs. In conclusion, inhibiting components of the adaptive immune response, like Tregs, are down regulated during a stress-induced activation of the adaptive immune response. In situations of chronic stress, this scenario might result in an exacerbation of inflammatory conditions such as autoimmune diseases. PMID- 20015596 TI - Five-year-olds' cortisol reactions to an attachment story completion task. AB - An earlier study showed that negative parent-child interactions elicit cortisol increases in 5-year-old children (Smeekens et al., 2007a). In the present study, we examined whether an "imaginary" parent-child interaction situation, in which the child used a parent and child doll to complete a set of more or less stressful story beginnings, also elicited cortisol increases in the children. In a sample of 89 five-year-old children (47 boys), an attachment story completion task (Verschueren and Marcoen, 1994) was administered during a school visit. Saliva samples to assess children's cortisol levels were collected right after arrival of the experimenter at the school (T0, baseline), immediately prior (T1, pretest) and 20min after (T2, posttest) the completion of the task. Hierarchical regression analysis indicated that children who displayed more insecure stories (i.e., stories depicting the parent as unavailable, unsupportive, or hostile) showed larger relative cortisol increases, but only if they had highly negative parent-child interactions 1 week earlier. Children who did not have negative interactions with the parent showed no cortisol increases when playing insecure stories. The results suggest that it is not the playing of insecure stories itself but perhaps the emotional memories of negative parent-child interactions triggered while playing insecure stories that elicit cortisol increases in children. This underscores the profound impact that negative interactions with parents may have on children's functioning and development, not only at the behavioural and emotional level, but also at the level of physiological arousal and regulation. Results also suggest that a secure attachment to one parent may act as a buffer against negative effects of low-quality interactions with the other parent, but more research is needed to more thoroughly test this assumption. PMID- 20015597 TI - Honey bee sting and venom offering active as well as passive immunization could reduce swine flu pandemic A (H1N1). PMID- 20015598 TI - Decreasing temporal lobe dose with five-field intensity-modulated radiotherapy for treatment of pituitary macroadenomas. AB - PURPOSE: To compare temporal lobe dose delivered by three pituitary macroadenoma irradiation techniques: three-field three-dimensional conformal radiotherapy (3D CRT), three-field intensity-modulated radiotherapy (3F IMRT), and a proposed novel alternative of five-field IMRT (5F IMRT). METHODS AND MATERIALS: Computed tomography-based external beam radiotherapy planning was performed for 15 pituitary macroadenoma patients treated at New York University between 2002 and 2007 using: 3D-CRT (two lateral, one midline superior anterior oblique [SAO] beams), 3F IMRT (same beam angles), and 5F IMRT (same beam angles with additional right SAO and left SAO beams). Prescription dose was 45 Gy. Target volumes were: gross tumor volume (GTV) = macroadenoma, clinical target volume (CTV) = GTV, and planning target volume = CTV + 0.5 cm. Structure contouring was performed by two radiation oncologists guided by an expert neuroradiologist. RESULTS: Five-field IMRT yielded significantly decreased temporal lobe dose delivery compared with 3D CRT and 3F IMRT. Temporal lobe sparing with 5F IMRT was most pronounced at intermediate doses: mean V25Gy (% of total temporal lobe volume receiving >=25 Gy) of 13% vs. 28% vs. 29% for right temporal lobe and 14% vs. 29% vs. 30% for left temporal lobe for 5F IMRT, 3D-CRT, and 3F IMRT, respectively (p < 10(-7) for 5F IMRT vs. 3D-CRT and 5F IMRT vs. 3F IMRT). Five-field IMRT plans did not compromise target coverage, exceed normal tissue dose constraints, or increase estimated brain integral dose. CONCLUSIONS: Five-field IMRT irradiation technique results in a statistically significant decrease in the dose to the temporal lobes and may thus help prevent neurocognitive sequelae in irradiated pituitary macroadenoma patients. PMID- 20015599 TI - Sustained release methylphenidate for the treatment of ADHD in amphetamine abusers: a pilot study. AB - The efficacy of stimulant treatment in patients with substance use disorders and comorbid attention deficit hyperactivity disorder (ADHD) has been tested for cocaine and alcohol dependence but so far no studies have been conducted in amphetamine dependent individuals. The present trial was a pilot study aiming to test the feasibility of treating amphetamine dependent patients with comorbid ADHD with central stimulant medication. The study was a double-blind, placebo controlled trial with parallel groups design comparing the efficacy of a fixed dose (72mg) of OROS methylphenidate (MPH) with placebo (PL) in reducing ADHD symptoms in currently abstinent adults with amphetamine dependence and ADHD. Twenty-four treatment seeking patients who met the DSM IV criteria for amphetamine dependence and ADHD were randomized to MPH/PL. The trial was conducted at an outpatient facility with twice weekly visits, measuring ADHD symptoms and drug use. Patients rated their ADHD symptoms on a weekly basis and provided supervised urine specimens for drug toxicology twice weekly. All patients participated in weekly sessions of a skills training programme. Both the groups significantly reduced their self-rated ADHD symptoms during the 12-week treatment but there was no difference between the two treatment arms. Drug use, both measured by urine toxicology and self-report did not differ between the groups. No difference was found between the two groups with regards to craving for amphetamine or in retention in treatment. Larger studies with higher doses combined with individual dosage and longer follow-up periods are warranted. PMID- 20015600 TI - Gabapentin use in hyperemesis gravidarum: a pilot study. AB - Among 7 subjects with hyperemesis gravidarum (HG), we found gabapentin therapy to be associated with mean reductions in nausea and emesis from Baseline to Days 12 14 of 80% and 94%, respectively. There have been 2 congenital defects among 7 exposed infants. Gabapentin may be effective in the treatment of HG. PMID- 20015601 TI - Sexual dysfunction and the ageing male. AB - Male sexuality in older age is an important issue but is not fully understood. This review aims to clarify the normal ageing process, the sexual behaviour of ageing men and the prevalence of sexual dysfunction. It identifies conditions affecting male sexuality in older age and highlights areas where more extensive research is required. PMID- 20015602 TI - The potential clinical value of FDG-PET for recurrent renal cell carcinoma. AB - PURPOSE: The clinical value of positron emission tomography (PET) using (18)F fluorodeoxyglucose (FDG) for follow-up or suspected recurrence of renal cell carcinoma (RCC) has not been fully evaluated. The purpose of this study was to assess the diagnostic performance of FDG-PET for postoperative assessment in patients with RCC. METHODS: We reviewed 28 scans in 23 patients who had undergone FDG-PET scans after surgery for RCC. Diagnostic accuracy of visually interpreted PET was evaluated based on final diagnoses obtained histologically or by clinical follow-up at least 6 months. Also, additional information over CT, influence on treatment decisions, and the accuracy of FDG uptake as a predictor of survival were assessed. RESULTS: Recurrence of renal carcinoma was histologically (n=15) or clinically (n=6) confirmed in 21 of 28 cases. Overall, the sensitivity, specificity, and diagnostic accuracy using FDG-PET were 81%, 71%, and 79%, respectively. In papillary RCC, the sensitivity was 100%; however, that was 75% in clear cell RCC in patient-basis. PET correctly detected local recurrence and metastases in all cases in the peritoneum, bone, muscle and adrenal gland. Additional information was obtained from scans in 6 cases (21%), which influenced therapeutic management in 3 cases (11%). Cumulative survival rates over 5 years in the PET-positive vs. the PET-negative group were 46% vs. 83%, respectively (p=0.17). CONCLUSIONS: FDG-PET would be useful for postoperative surveillance in patients with RCC, although its impact on treatment decisions may be limited. Further investigations are necessary to conclude whether PET has a prognostic value. PMID- 20015603 TI - Evaluation of the therapeutic efficacy of high-intensity focused ultrasound ablation of hepatocellular carcinoma by three-dimensional sonography with a perflubutane-based contrast agent. AB - OBJECTIVE: We performed contrast-enhanced three-dimensional sonography (CE 3D US) with a perflubutane-based contrast agent to immediately evaluate the completeness of ablation of small hepatocellular carcinoma (HCC) lesions by extracorporeal high-intensity focused ultrasound (HIFU). SUBJECTS AND METHODS: Twenty-one HCC lesions were treated by a single ultrasound-guided HIFU ablation session, and CE 3D US was performed before, immediately after, and 1 week, and 1 month after HIFU, and contrast-enhanced CT (CE CT) or contrast-enhanced MRI (CE MRI) was performed before HIFU, 1 week and 1 month after HIFU, and during the follow-up period. RESULTS: Immediately and 1 month after HIFU, 17 lesions were evaluated as adequately ablated by CE 3D US, and the other 4 lesions as residual tumors. One month after HIFU, 18 were evaluated as adequately ablated by CE CT or CE MRI, and the other 3 as residual tumors. The evaluation by CE 3D US immediately after HIFU and by CE CT or CE MRI 1 month after HIFU was concordant with 20 lesions. The kappa value for agreement between the findings of CE 3D US and other modalities by two blinded observers was 0.83. When the 1-month CE CT or CE MRI findings were used as the reference standard, the sensitivity, specificity, and accuracy of CE 3D US immediately after HIFU for the diagnosis of the adequate ablation were 100%, 75%, and 95%, respectively. CONCLUSION: CE 3D US appears to be a useful method for immediate evaluation of therapeutic efficacy of HIFU ablation of HCC lesions. PMID- 20015604 TI - Proteomic identification of human serum biomarkers in diabetes mellitus type 2. AB - Discovery of protein biomarkers in different diseases is an important area of research in the field of proteomics. We have described the levels of protein biomarkers specific to diabetes mellitus type 2 in the local population of Pakistan using proteomic technology. Type 2 diabetic patients, age and sex matched normal healthy controls were recruited from Sheikh Zayed Hospital, Lahore, Pakistan. Plasma proteins were analysed by 2D liquid chromatographic system in which samples were initially fractionated by chromatofocusing and the selected fractions were further analysed by reverse-phase high performance liquid chromatography. The proteins which showed variation between test and control samples were identified by MALDI-TOF analysis. All the samples belonging to the control and diabetic groups were then analyzed by ELISA and estimated four proteins which were found to vary. Levels of apolipoprotein A-I was found to decrease by -6.4% while apolipoprotein E, leptin and C reactive protein (CRP) were increased by +802, +842 and +872%, respectively, in the diabetic patients as compared to the controls. The discovery of these marker proteins might thus provide an adjunctive method for early detection of risk for this disease. PMID- 20015605 TI - A randomized experimental study of gender-responsive substance abuse treatment for women in prison. AB - This experimental pilot study compared postrelease outcomes for 115 women who participated in prison-based substance abuse treatment. Women were randomized to a gender-responsive treatment (GRT) program using manualized curricula (Helping Women Recover and Beyond Trauma) or a standard prison-based therapeutic community. Data were collected from the participants at prison program entry and 6 and 12 months after release. Bivariate and multivariate analyses were conducted. Results indicate that both groups improved in psychological well being; however, GRT participants had greater reductions in drug use, were more likely to remain in residential aftercare longer (2.6 vs. 1.8 months, p < .05), and were less likely to have been reincarcerated within 12 months after parole (31% vs. 45%, respectively; a 67% reduction in odds for the experimental group, p < .05). Findings show the beneficial effects of treatment components oriented toward women's needs and support the integration of GRT in prison programs for women. PMID- 20015606 TI - Structural-level factors affecting implementation of the methadone maintenance therapy program in China. AB - This study identifies structural-level factors influencing methadone maintenance therapy (MMT) program implementation in China. Twenty-eight service providers and 560 randomly selected clients from 28 MMT clinics in the study area underwent face-to-face interviews. Number of clients, retention rate, coverage, and structural-level factors were collected from a survey of service providers. Individual-level factors and self-reported illicit drug use information were obtained from clients. Urine specimens were collected from the client participants to test for heroin use. Clinics affiliated with the local Centers for Disease Control and Prevention (CDC) had more clients, higher retention rates, and broader coverage than those not affiliated with the CDC. Longer operating hours, incentive for compliant clients, and comprehensive services were positively associated with client recruitment and coverage. Comprehensive services and incentives for compliant clients were negatively associated with concurrent illicit drug use. Comprehensive services should be incorporated into the MMT program. Extended operating hours and incentives for compliant clients should be implemented. PMID- 20015607 TI - A review of computer-based alcohol problem services designed for the general public. AB - This review summarizes the literature on computer-based drinking assessment and intervention programs evaluated using members of the general public. The primary aim was to summarize the demand, usage, and effectiveness of these services. A systematic search of the literature identified seven online drinking assessments and eight computerized interventions that were evaluated using members of the general public. Internet assessment users tend to be in their early 30s, are more often male, tend to be at risk for or are experiencing alcohol-related problems, more fully explore assessment sites, and are more likely to enroll in interventions linked to these sites when their drinking problem is more severe. Although dropout from computer-based interventions is often very high and treatment models vary widely, program completers appear to show improvements. PMID- 20015608 TI - Smoking cessation interventions among individuals in methadone maintenance: a brief review. AB - BACKGROUND: Individuals in methadone maintenance treatment are motivated to quit smoking and are interested in smoking cessation treatment. However, few studies have assessed the efficacy of smoking cessation treatment for individuals undergoing methadone maintenance. The purpose of this article is to review interventions for tobacco use cessation among individuals in methadone maintenance with a particular emphasis on the components of such interventions and their effect on smoking cessation/reduction and drug use. METHODS: A comprehensive search of six databases in June 2008 retrieved 584 research studies, which addressed smoking cessation interventions among individuals in substance use treatment. Of the retrieved articles, eight studies addressed smoking cessation among individuals undergoing methadone maintenance treatment. These studies were examined for effect of intervention on smoking cessation/reduction, gender differences in treatment outcomes, and the effect of the smoking cessation intervention on substance use behaviors. RESULTS: Few studies demonstrated successful smoking abstinence among individuals in methadone maintenance treatment. Most interventions were associated with significant smoking reduction from baseline. Few studies assessed differences between men and women in smoking cessation treatment outcome. Smoking cessation treatment was not associated with increased substance use. CONCLUSIONS: To date, interventions among individuals in methadone treatment have been largely unsuccessful in achieving sustained smoking abstinence. However, smoking cessation treatment does not worsen substance use. Future studies are necessary to determine intervention designs and components that can enhance smoking cessation among individuals in methadone maintenance. PMID- 20015609 TI - Recruited bone marrow cells expressing the EP3 prostaglandin E receptor subtype enhance angiogenesis during chronic inflammation. AB - Chronic inflammation, which is characterized by the proliferation of granulation tissues, is known to be regulated by angiogenesis. Recent results suggest that bone marrow-derived (BM-derived) hematopoietic cells regulate angiogenesis in vivo. We previously reported that the angiogenesis occurring during chronic inflammation is enhanced in response to the endogenous prostaglandins (PGs) derived from an inducible cyclooxygenase-2 (COX-2). In the present study, we examined the role of BM-derived cells expressing an E-type PG receptor subtype, EP3, in sponge-induced angiogenesis. The replacement of wild-type (WT) BM with BM cells (BMCs) from green fluorescent protein (GFP) transgenic mice revealed that the formation of granulation tissue around the sponge implants developed via the recruitment of BMCs. This recruitment was enhanced by topical injections of vascular endothelial growth factor (VEGF)-A, and a VEGF-dependent increase in the recruitment of BMCs was inhibited by a COX-2 inhibitor, celecoxib. FACS analysis of the granulation tissues after treatment with collagenase revealed that the Mac 1-positive macrophage fraction was enhanced by topical injections of VEGF-A, and that this increased recruitment of Mac-1-positive BMCs was inhibited by celecoxib. Selective knockdown of EP3 performed by BM transplantation with BMCs isolated from EP3 knockout (EP3) mice reduced sponge-induced angiogenesis, as estimated by mean vascular number and CD31 expression in the granulation tissues. This reduction in angiogenesis in EP3(-/-) BM chimeric mice was accompanied by reductions in the recruitment of BMCs, especially of Mac-1-positive cells and Gr 1-positive cells. These results indicate that the recruited bone marrow cells that express the EP3 receptor have a significant role in enhancing angiogenesis during chronic proliferative inflammation. PMID- 20015610 TI - [Obliterating arterial disease of the legs in women]. AB - The widespread use of the ankle-brachial index (ABI) enabled to reveal the high prevalence of lower extremities peripheral arterial disease (PAD) in women, even higher than in men in several series. Nonetheless, the use of a same ABI threshold for men and women may overestimate the prevalence of subclinical PAD in women. Conversely, the clinical manifestations of PAD are more often atypical in women, with as a result, a delayed diagnosis and management. The level of association between cardiovascular disease risk factors and PAD is similar in both genders. To date, there is no evidence for any association between PAD and different hormonal events in women. Despite an inverse association between hormonal replacement therapy (HRT) after menopause and PAD in epidemiological studies, the trials failed to evidence any protective effect of HRT to prevent PAD. Women with PAD are advised to discontinue HRT in case of limb revascularization. The long-term local and functional prognosis of PAD seems more severe in women, but this is partly explained by a delayed onset of the disease, occurring at a later age. Similarly, the general cardiovascular prognosis PAD is also poorer in women than in men. PMID- 20015611 TI - [Aortic disease in women]. AB - Abdominal aortic disease in women is associated with atheromatous processes much more often than with inflammatory arterial diseases (such as Takayasu or Horton). Intramural hematomas and atheromatous ulcers of the aorta, albeit rare, affect as many women as men and have variable outcomes: some are resolved with heparin treatment, and others are fatal. Atherosclerotic stenoses and occlusions limited to the level of the aortoiliac bifurcation are increasingly prevalent and difficult to treat in young women, in part because of their addiction to smoking. After being more or less ignored for a long time, the particularity of atheromatous abdominal aortic aneurysms (AAA) in women is now being recognized. AAA in women are less frequent than in men, but grow faster and have a higher risk of rupture; moreover, cardiovascular disease is generally detected and managed less often in women. Overall mortality from AAA in women is similar to that of breast cancer. The two major risk factors for AAA are, as in men, a direct family history of AAA and smoking. The diagnostic standards for AAA, the criteria for defining progression, and the indications for surgery are probably not the same as in men, and the smaller initial caliber of women's aortas must be taken into account. Most guidelines today recommend ultrasound screening for AAA for women older than 50 years with a family history (in a 1(st)degree relative), women aged 60-75 years who are hypertensive or smoke, and smokers older than 75 years without serious comorbidity and with a life expectancy essentially normal for their age. Monitoring patients with a small AAA (anteroposterior diameter < 40-45 mm) must not be limited only to the aneurysm, but must also include comprehensive management to eliminate modifiable risk factors and thus to reduce cardiovascular and surgical risk (by better preparing the patient for this possibility) as well as to slow the progression of the aneurysm and decrease the risk of its rupture. PMID- 20015612 TI - A study on the use of imputation methods for experimentation with Radial Basis Function Network classifiers handling missing attribute values: the good synergy between RBFNs and EventCovering method. AB - The presence of Missing Values in a data set can affect the performance of a classifier constructed using that data set as a training sample. Several methods have been proposed to treat missing data and the one used more frequently is the imputation of the Missing Values of an instance. In this paper, we analyze the improvement of performance on Radial Basis Function Networks by means of the use of several imputation methods in the classification task with missing values. The study has been conducted using data sets with real Missing Values, and data sets with artificial Missing Values. The results obtained show that EventCovering offers a very good synergy with Radial Basis Function Networks. It allows us to overcome the negative impact of the presence of Missing Values to a certain degree. PMID- 20015613 TI - Drugs and autoimmunity--a contemporary review and mechanistic approach. AB - Drug-induced autoimmunity is an idiosyncratic, non-IgE immune related drug reaction. Interestingly, although many drugs have been reported to induce autoantibodies, only a few have a definitive association with drug-induced autoimmune disease. The prototype disease is drug-induced lupus and the typical drug for drug-induced lupus is minocycline. The production of autoantibodies and the induction of symptoms in drug-induced lupus results from a variety of mechanisms, which can include suppression of central or peripheral tolerance, alteration of gene transcription in T and B cells, abnormal cytokine and/or cytokine receptor balance and function, chromatin structure modification and antigen modification. Multiple mechanisms may apply for different drugs, and understanding the pharmacological actions of these agents helps us decipher the etiology. For example, DNA hypomethylation may occur with hydralazine, which leads to increased transcription, increased LFA-1, the generation of autoreactive T cells and a breakdown in peripheral tolerance. Frequently, more than one pathway may be involved. Interestingly, most patients with newly formed autoantibodies resulting from drugs do not develop clinical disease. Nonetheless, the explosion in the use of biological modifiers has been associated with production of autoantibodies, an observation that illustrates the complex nature of these interactions, in that these agents are frequently used to treat autoimmunity, yet may produce autoimmune diseases themselves. PMID- 20015614 TI - Surgical management of bisphosphonate induced osteonecrosis of the jaws. AB - Recent literature contains numerous articles on osteonecrosis of the jaws (ONJ) associated with bisphosphonate treatment (BPT), with most advocating a conservative approach to management. A prospective study was designed to review the surgical management of cases of ONJ that did not respond to conservative management. Forty patients, referred with ONJ that did not respond to conservative management, were treated surgically and followed up for 6 months to 4 years. Four patients were taking i.v. BPT as part of their bone cancer management and 16 were taking oral BPT for osteoporosis. The surgical management of ONJ involved antibiotic therapy, surgical debridement of all necrotic bone and tension-free primary closure. All 40 cases healed uneventfully with no wound breakdown during follow-up. Most of the literature supports the conservative management of ONJ, but the condition leaves the patient debilitated. Many cases do not respond to conservative management and the infection and bone destruction is progressive. The conservative management of ONJ is to be supported, but this prospective study has shown that those cases that do not respond may be managed surgically. It should be recognized that while the results of this paper are encouraging, some cases will be resistant to all treatments. PMID- 20015615 TI - Evidences for pharmacokinetic interaction of riluzole and topiramate with pilocarpine in pilocarpine-induced seizures in rats. AB - In this study we investigated the effectiveness of two antiepileptic drugs: riluzole and topiramate against pilocarpine-induced seizures, which are considered to be a model of intractable epilepsy commonly used to investigate the antiepileptic effect of drugs and mechanisms of epileptogenesis. Seizures and status epilepticus were induced by pilocarpine in adult male Wistar rats. Riluzole (1-4mg/kg) administered intraperitoneally before pilocarpine dose dependently protected rats against seizures with the anticonvulsant ED(50) value (50% effective anticonvulsant dose) of 1.8 (1.3-2.6)mg/kg. In contrast, riluzole at 8 and 12mg/kg administered after the onset of pilocarpine-induced seizures affected neither status epilepticus nor mortality of rats. Topiramate significantly enhanced convulsive action of pilocarpine, lowering the convulsant CD(50) value (50% effective convulsant dose) of pilocarpine from 350.8 (329.2 373.8) to 246.4 (218.6-278.2)mg/kg. Riluzole (4mg/kg) lowered plasma and brain concentration of pilocarpine administered at a dose of 400mg/kg from 168.0+/-8.6 to 75.3+/-19.9microg/ml and from 193.7+/-6.6 to 97.0+/-26.1microg/g, respectively. Topiramate (200mg/kg) increased plasma and brain concentration of pilocarpine administered at a dose of 300mg/kg from 78.1+/-2.9 to 106.0+/ 6.8microg/ml and from 138.4+/-5.0 to 155.2+/-5.1microg/g, respectively. It seems that both anticonvulsant effect exerted by riluzole and proconvulsant effect exerted by topiramate in pilocarpine model of seizures are due to a pharmacokinetic interaction. Therefore, we postulate that the concentration of pilocarpine should be measured routinely whenever the anticonvulsant effect of drugs is determined in the pilocarpine model of seizures. PMID- 20015616 TI - Distribution of EFHC1 or Myoclonin 1 in mouse neural structures. AB - EFHC1, a gene mutated in juvenile myoclonic epilepsy, encodes EFHC1, a protein with three DM10 domains and one EF-hand motif. We recently demonstrated that this molecule is a microtubule-associated protein (MAP) implicated in neuronal migration. Because some controversies persist about the precise localization in the CNS, we studied the neuroanatomical distribution of EFHC1 in mature and developing mouse brain. In the adult, low mRNA expression was detected in several brain structures such as cortex, striatum, hippocampus and cerebellum. At E16, EFHC1 mRNA was shown to be expressed in cortex and not only in cells lining ventricles. Using a purified polyclonal antibody, EFHC1 staining was observed in all cortical layers, in piriform cortex, in hippocampus and in Purkinje cells of cerebellum. In the cortex, the majority of EFHC1 positive cells correspond to neurons, however some glial cells were also stained. In agreement with a previous study, we demonstrated strong EFHC1 expression in cilia of ependymal cells lining cerebral ventricles. Moreover, at E16, the protein was observed at the borders of brain ventricles, in choroid plexus, but also, although to a lesser extent, in piriform and neocortex. In these latter structures, the pattern of expression seems to correspond to the extensions of the radial glia fibers as demonstrated by BLBP immunostaining. Finally, we confirmed that EFHC1 was also expressed and co-localized with the mitotic spindle of neural stem cells. These results confirm that EFHC1 is a protein with a likely low expression level in mouse brain but detectable both in adult and embryonic brain supporting our previous data and hypothesis that EFHC1 could play an important role during brain development. As discussed, this opens the door to a new conceptual approach viewing some idiopathic generalized epilepsies as developmental diseases instead of classical channelopathies. PMID- 20015617 TI - Effects of levetiracetam and valproic acid monotherapy on sex-steroid hormones in prepubertal children--results from a pilot study. AB - The influence of levetiracetam (LEV) and valproic acid (VPA) monotherapy on sex steroid hormone profile was investigated in thirty prepubertal children. VPA treated children showed greatest androstendione concentrations when compared to LEV treated children (p=0.016) and to controls (p=0.011). All other reproductive endocrine hormones were similar among groups. In conclusion, LEV does not seem to induce changes in reproductive endocrine functions as well as clinically relevant endocrine side effects in prepubertal children. PMID- 20015618 TI - Activity of aminocandin (IP960; HMR3270) compared with amphotericin B, itraconazole, caspofungin and micafungin in neutropenic murine models of disseminated infection caused by itraconazole-susceptible and -resistant strains of Aspergillus fumigatus. AB - Aminocandin (IP960; HMR3270; NXL201) is a new echinocandin with broad-spectrum in vitro activity against Aspergillus and Candida spp. We compared the activity of aminocandin with that of amphotericin B (AmB), itraconazole (ITC) and caspofungin (CAS) in murine models of disseminated aspergillosis against three strains of A. fumigatus, two of which were fully susceptible (AF293 and A1163) and one was resistant to ITC (AF91). Mice were rendered temporarily neutropenic or persistently neutropenic with cyclophosphamide and were infected intravenously 3 days later. Temporarily neutropenic mice were treated with either intraperitoneal (i.p.) AmB (5mg/kg/dose), oral (p.o.) ITC (25mg/kg/dose), intravenous (i.v.) aminocandin (0.25-10mg/kg/dose), i.p. aminocandin (1mg/kg/dose) or solvent control for 9 days. Mice were euthanised 11 days post infection and the kidneys and liver were removed for quantitative culture. Following infection with AF293, only aminocandin 5mg/kg i.v. yielded 100% survival. Aminocandin 1mg/kg i.v., AmB 5mg/kg i.p. or ITC 25mg/kg p.o. were equivalent (P>0.05). Aminocandin 5mg/kg was superior to aminocandin 0.25mg/kg (P<0.0001) as well as all controls (P<0.0001) in reducing mortality. Following infection with AF91, only aminocandin at 5mg/kg and 1mg/kg i.v. yielded 100% survival, which was superior to ITC, aminocandin 0.25mg/kg and controls (all P<0.0001). In the persistently neutropenic model with A1163, aminocandin, CAS and micafungin (2-10mg/kg) were all effective at prolonging survival, with some impact on reducing culture burdens, even with alternate-day dosing (4mg/kg). The only fungicidal regimen was aminocandin 5mg/kg, which sterilised 40% and 50% of mice following infection with AF293 and AF91, respectively. Aminocandin at doses of > or =1mg/kg is highly effective in reducing mortality and organ burden in disseminated infection caused by ITC susceptible and -resistant A. fumigatus. PMID- 20015620 TI - A mouse model of anxiety molecularly characterized by altered protein networks in the brain proteome. AB - Recently, several attempts have been made to describe changes related to certain anxiety states in the proteome of experimental animal models. However, these studies are restricted by limitations regarding the number and correct identification of separated proteins. Moreover, the application of a systems biology approach to discover the molecular mechanisms of anxiety requires genetically homogenous inbred animal models. Therefore, we developed a novel mouse model of anxiety using a combination of crossbreeding (inbred for 35 generations) and behavioral selection. We found significant changes in 82 proteins in the total brain proteome compared to the control proteome. Thirty four of these proteins had been previously identified in other anxiety, depression or repeated psychosocial stress studies. The identified proteins are associated with different cellular functions, including synaptic transmission, metabolism, proteolysis, protein biosynthesis and folding, cytoskeletal proteins, brain development and neurogenesis, oxidative stress, signal transduction. Our proteomics data suggest that alterations in serotonin receptor-associated proteins, in the carbohydrate metabolism, in the cellular redox system and in synaptic docking are all involved in anxiety. PMID- 20015619 TI - Potential anxiogenic effects of cannabinoid CB1 receptor antagonists/inverse agonists in rats: comparisons between AM4113, AM251, and the benzodiazepine inverse agonist FG-7142. AB - Cannabinoid CB1 inverse agonists suppress food-motivated behaviors, but may also induce psychiatric effects such as depression and anxiety. To evaluate behaviors potentially related to anxiety, the present experiments assessed the CB1 inverse agonist AM251 (2.0-8.0mg/kg), the CB1 antagonist AM4113 (3.0-12.0mg/kg), and the benzodiazepine inverse agonist FG-7142 (10.0-20.0mg/kg), using the open field test and the elevated plus maze. Although all three drugs affected open field behavior, these effects were largely due to actions on locomotion. In the elevated plus maze, FG-7142 and AM251 both produced anxiogenic effects. FG-7142 and AM251 also significantly increased c-Fos activity in the amygdala and nucleus accumbens shell. In contrast, AM4113 failed to affect performance in the plus maze, and did not induce c-Fos immunoreactivity. The weak effects of AM4113 are consistent with biochemical data showing that AM4113 induces little or no intrinsic cellular activity. This research may lead to the development of novel appetite suppressants with reduced anxiogenic effects. PMID- 20015621 TI - Electrochemical investigation of tryptophan at gold nanoparticles modified electrode in the presence of sodium dodecylbenzene sulfonate. AB - Glassy carbon electrode modified with gold nanoparticles was prepared using electrodeposition at constant potential of -0.20V, and characterized with scanning electron microscopy and electrochemical techniques. Voltammetry was employed to study the electrochemical behaviors of tryptophan at the Au nanoparticles modified electrode in the presence of sodium dodecylbenzene sulfonate. The results showed that the Au nanoparticles modified electrode had good performance for the electrochemical oxidation of tryptophan. Compared with the unmodified electrode, the Au nanoparticles modified electrode improved the adsorption of tryptophan on the nanointerface and amplified its current response. The oxidation peak potential showed a negative shift of 50mV in the presence of sodium dodecylbenzene sulfonate indicating that the electron transfer between the electrode and bulk solution of tryptophan was facilitated. Experimental parameters for tryptophan determination, such as deposition time, pH value, and accumulation conditions have been optimized. The oxidation peak current was linearly dependent on the tryptophan concentration and a calibration curve was obtained in the concentration range of 9.0x10(-8)molL(-1) to 5.0x10(-5)molL(-1) with detection limit of 8.0x10(-8)molL(-1) (S/N=3). PMID- 20015622 TI - Biogenic synthesis of Au and Ag nanoparticles by Indian propolis and its constituents. AB - In an attempt to find natural, environmentally benign, green-chemical agents for the synthesis of metal nanoparticles, we have demonstrated for the first time the excellent efficiency of ethanol and water extracts of a natural, non-toxic material, Indian propolis and two of its chemical constituents, pinocembrin and galangin in the rapid synthesis of stable Ag and Au nanoparticles having wide spectrum of fascinating morphologies. Both of these two extracts were found to be extremely efficient in the synthesis of Ag and Au nanoparticles under alkaline condition. For a given metal ion precursor, the kinetics of particle synthesis were remarkably similar in all the cases, as it is evident from the absorption spectra monitored over time. Moreover they exhibited similar redox behavior under alkaline condition (pH approximately 10.62). The efficiency of the ethanol and water extracts of Indian propolis towards Ag and Au nanoparticles synthesis was compared with that of naturally occurring hydroxyflavonoids, pinocembrin and galangin isolated from Indian propolis; which are equally efficient in the rapid synthesis of Ag and Au nanoparticles and stabilization of the resultant particles. PMID- 20015623 TI - Mobile phone use within an oncology department. PMID- 20015624 TI - Sickle cell crisis in a patient receiving capecitabine chemotherapy. PMID- 20015625 TI - Biodiversity of streptomycetes isolated from a succession sequence at a post mining site and their evidence in Miocene lacustrine sediment. AB - The genetic diversity of streptomycetes in colliery spoil heaps (Sokolov, Czech Republic) was investigated by restriction pattern analysis of 16S-internal transcribed spacer rDNA and 16S sequences. We sampled freshly excavated Miocene sediment (17-19-million-year-old) and four sites of primary succession (initial, early, middle, and late stages; aged 1-44 years) on the same sediment. Active bacteria were present even in fresh Miocene sediment, and the relative proportion of actinomycetes among total bacterial and their genetic diversity increased significantly with the age of the sampling site. The replacement of pioneer species by late succession species during succession was observed. Plate assays of Streptomyces strains revealed 27% antibiotic-producing strains. Screening for nonribosomal peptide synthases and type I polyketide synthases systems suggested that 90% and 55% streptomycetes, respectively, are putative producers of biologically active compounds. The frequencies of tetracycline-, amoxicillin-, and chloramphenicol-resistant streptomycetes were 6%, 9%, and 15%, respectively. These findings document the occurrence of genetic elements encoding antibiotic resistance genes and the production of antibiotics by streptomycetes located in pristine environments. Our results indicate key roles for ancient streptomycetes related to S. microflavus, S. spororaveus, and S. flavofuscus in pioneering community development in freshly excavated substrates. PMID- 20015626 TI - Coordination to gallium(III) strongly enhances the potency of 2-pyridineformamide thiosemicarbazones against Cryptococcus opportunistic fungi. AB - A family of 2-pyridineformamide-derived thiosemicarbazones and their gallium(III) complexes were tested against several isolates of pathogenic Cryptococcus strains. On complexation the antifungal activity significantly increases, suggesting coordination to gallium(III) to be an interesting strategy of antifungal dose reduction. PMID- 20015627 TI - Characterization of a cytochrome c gene located at the gene cluster for chlorate respiration in Ideonella dechloratans. AB - Anaerobic chlorate respiration requires electron transport from the bacterial inner membrane to the soluble periplasmic chlorate reductase. We have recently demonstrated that soluble c cytochromes function as electron carriers for chlorate reduction in Ideonella dechloratans (Smedja Backlund et al. 2009). In the present work, we describe a gene encoding soluble c-type cytochrome [cyt; GenBank ID: EU768872] located close to the gene cluster for chlorate reduction in I. dechloratans. The predicted amino acid sequence does not match any of the peptide masses or partial sequences obtained earlier from periplasmic c cytochromes. The gene, without the predicted signal sequence, was expressed heterologously in E. coli and the recombinant protein was purified, refolded and reconstituted with heme. The reconstituted protein shows spectral properties characteristic for c cytochromes, with an absorption maximum at 553 nm for the alpha band in the reduced state. Pyridine hemochrome analysis demonstrates the formation of covalently bound heme. PMID- 20015628 TI - Functional characterization of KanP, a methyltransferase from the kanamycin biosynthetic gene cluster of Streptomyces kanamyceticus. AB - KanP, a putative methyltransferase, is located in the kanamycin biosynthetic gene cluster of Streptomyces kanamyceticus ATCC12853. Amino acid sequence analysis of KanP revealed the presence of S-adenosyl-L-methionine binding motifs, which are present in other O-methyltransferases. The kanP gene was expressed in Escherichia coli BL21 (DE3) to generate the E. coli KANP recombinant strain. The conversion of external quercetin to methylated quercetin in the culture extract of E. coli KANP proved the function of kanP as S-adenosyl-L-methionine-dependent methyltransferase. This is the first report concerning the identification of an O methyltransferase gene from the kanamycin gene cluster. The resistant activity assay and RT-PCR analysis demonstrated the leeway for obtaining methylated kanamycin derivatives from the wild-type strain of kanamycin producer. PMID- 20015629 TI - Isolation of heterotrophic bacteria from Palk Bay sediments showing heavy metal tolerance and antibiotic production. AB - Analysis of culturable and unculturable bacteria and their potential bioactive compounds from Palk Bay is yet to be explored. The present study for the first time characterizes the culturable bacteria from Palk Bay sediment using 16S rRNA gene sequencing. The characterized bacteria were also screened for antibacterial activity against human and aquaculture pathogens. In the 16S rRNA gene sequence analysis characterized that most of the bacteria were affiliated to members of Firmicutes and less with Gammaproteobacteria, Actinobacteria and Alphaproteobacteria. A high portion of (39%) of the bacteria showed antibacterial activity against both Gram positive and Gram negative test strains. The antibiotics from the strain S6-05 were partially purified using solvent extraction followed by TLC and silica column and further characterized by IR analysis. Two active principles A and B showed difference in the activity against Gram positive and Gram negative pathogens. But in the synergistic application they showed excellent activity against all the test pathogens. This study provides the first evidence on the existence of certain Bacillus species in the marine environment, namely Bacillus arsenicus, Bacillus indicus, Bacillus boroniphilus, Bacillus cibi and Bacillus niabensis which also had antibacterial activity. Several of the isolates exhibited tolerance to arsenic and boron to a new level of 25 and 100 mM, respectively. The current study reveals the fact that a great deal remains in the bacterial diversity of Palk Bay region. PMID- 20015630 TI - Chest low-dose computed tomography in neutropenic acute myeloid leukaemia patients. AB - BACKGROUND: We aimed to compare chest low-dose computed tomography (LDCT) with chest radiography (CXR) in the assessment of febrile acute myeloid leukaemia neutropenic patients. METHODS: A prospective non-randomized study was carried out between 30 May, 2003 and 3 June, 2004 in consecutive neutropenic patients who required imaging of the thorax and were treated for acute myeloid leukaemia. Each patient had a baseline 2-view chest radiograph followed by LDCT. Both the CXR and the LDCT studies were blindly and independently reviewed by two chest radiologists. RESULTS: Forty patients were enrolled: 24 male and 16 female, mean age 53.5 years (range 18-83) and an average neutrophil count of 0.78 x 10(9)/L. Patients had CXR within a mean of 40 min from the LDCT. Overall, 31 (77.5%) of 40 CXR were abnormal, whereas LDCT detected abnormalities in 38 (95%) of 40 patients. LDCT demonstrated three times the number of lung nodules as CXR and twice as many ground-glass opacities. Lung consolidation was detected similarly using both techniques, but LDCT demonstrated more extensive and multi-focal consolidation. The majority of nodules detected only on LDCT were subcentimetre in diameter. The additional information provided by LDCT led to an alteration in the clinical management of 11 (27.5%) of 40 patients. CONCLUSION: LDCT is a useful tool in the initial investigation of suspected pulmonary complication in neutropenic patients. This is supported by the additional information it provides to the CXR with reduced radiation when compared to conventional CT. PMID- 20015631 TI - Potent antiplatelet activity of sesamol in an in vitro and in vivo model: pivotal roles of cyclic AMP and p38 mitogen-activated protein kinase. AB - Sesamol is a potent phenolic antioxidant which possesses antimutagenic, antihepatotoxic and antiaging properties. Platelet activation is relevant to a variety of acute thrombotic events and coronary heart diseases. There have been few studies on the effect of sesamol on platelets. Therefore, the aim of this study was to systematically examine the detailed mechanisms of sesamol in preventing platelet activation in vitro and in vivo. Sesamol (2.5-5 MUM) exhibited more potent activity of inhibiting platelet aggregation stimulated by collagen than other agonists. Sesamol inhibited collagen-stimulated platelet activation accompanied by [Ca(2+)](i) mobilization, thromboxane A(2) (TxA(2)) formation, and phospholipase C (PLC)gamma2, protein kinase C (PKC) and mitogen activated protein kinase (MAPK) phosphorylation in washed platelets. Sesamol markedly increased cAMP and cGMP levels, endothelial nitric oxide synthase (eNOS) expression and NO release, as well as vasodilator-stimulated phosphoprotein (VASP) phosphorylation. SQ22536, an inhibitor of adenylate cyclase, markedly reversed the sesamol-mediated inhibitory effects on platelet aggregation and p38 MAPK phosphorylation, and sesamol-mediated stimulatory effects on VASP and eNOS phosphorylation, and NO release. Sesamol also reduced hydroxyl radical (OH(?)) formation in platelets. In an in vivo study, sesamol (5 mg/kg) significantly prolonged platelet plug formation in mice. The most important findings of this study demonstrate for the first time that sesamol possesses potent antiplatelet activity, which may involve activation of the cAMP-eNOS/NO-cGMP pathway, resulting in inhibition of the PLCgamma2-PKC-p38 MAPK-TxA(2) cascade, and, finally, inhibition of platelet aggregation. Sesamol treatment may represent a novel approach to lowering the risk of or improving function in thromboembolism related disorders. PMID- 20015632 TI - Applying decision-making tools to national e-waste recycling policy: an example of Analytic Hierarchy Process. AB - As policy making is in essence a process of discussion, decision-making tools have in many cases been proposed to resolve the differences of opinion among the different parties. In our project that sought to promote a country's performance in recycling, we used the Analytic Hierarchy Process (AHP) to evaluate the possibilities and determine the priority of the addition of new mandatory recycled waste, also referred to as Due Recycled Wastes, from candidate waste appliances. The evaluation process started with the collection of data based on telephone interviews and field investigations to understand the behavior of consumers as well as their overall opinions regarding the disposal of certain waste appliances. With the data serving as background information, the research team then implemented the Analytic Hierarchy Process using the information that formed an incomplete hierarchy structure in order to determine the priority for recycling. Since the number of objects to be evaluated exceeded the number that the AHP researchers had suggested, we reclassified the objects into four groups and added one more level of pair-wise comparisons, which substantially reduced the inconsistency in the judgment of the AHP participants. The project was found to serve as a flexible and achievable application of AHP to the environmental policy-making process. In addition, based on the project's outcomes derived from the project as a whole, the research team drew conclusions regarding the government's need to take back 15 of the items evaluated, and suggested instruments that could be used or recycling regulations that could be changed in the future. Further analysis on the top three items recommended by the results of the evaluation for recycling, namely, Compact Disks, Cellular Phones and Computer Keyboards, was then conducted to clarify their concrete feasibility. After the trial period for recycling ordered by the Taiwan Environmental Protection Administration, only Computer Keyboards were referred to as the new Due Recycled Waste and their producers started to take responsibility for recycling them from 2007 onwards. PMID- 20015633 TI - Antitumour activity of NK012, SN-38-incorporating polymeric micelles, in hypovascular orthotopic pancreatic tumour. AB - Human pancreatic cancer is refractory to chemotherapy partly because of blockage to penetration of anticancer agents. This issue must be taken into account particularly for the drug delivery system (DDS). The aim of the present study is to investigate how NK012 (SN-38-incorporating polymeric micelles) categorised as DDS exerts its antitumour effect in an orthotopic pancreatic tumour model compared with gemcitabine and irinotecan hydrochloride (CPT-11), a low-molecular weight prodrug of a 7-ethyl-10-hydroxy-camptothecin (SN-38). The maximum tolerated doses (MTDs) of NK012 (30 mg/kg/d), CPT-11 (66.7 mg/kg/d) and gemcitabine (16.5mg/kg/d) were administered to mice bearing human pancreatic cancer cell (SUIT-2) xenografts implanted orthotopically. Antitumour effects of these compounds were evaluated. Drug distribution within the tumour was examined by fluorescence microscopy and high performance liquid chromatography (HPLC). NK012 exerted potent antitumour effects compared with CPT-11 and gemcitabine. A high concentration of NK012 and SN-38 released from NK012 had been observed until 192h. On the other hand, SN-38 converted from CPT-11 was detected only 1h postinjection. Fluorescence from NK012 was detected up to 48h, whereas that from CPT-11 almost disappeared by 24h postinjection. NK012 appeared to exert potent antitumour activity against intractable stroma-rich orthotopic pancreatic tumour xenografts due to its sufficient accumulation followed by the effective sustained release of SN-38 from NK012. PMID- 20015634 TI - Is early stage endometrial cancer safely treated by laparoscopy? Complications of a multicenter study and review of recent literature. AB - BACKGROUND: To compare the complications after total laparoscopic hysterectomy (LPS) and abdominal hysterectomy with lymphadenectomy (LPT) for early stage endometrial cancer in a series of 226 women and to assess the disease-free survival and the recurrence rate. PATIENTS AND METHODS: Two hundred and twenty six patients with clinical stage I endometrial cancer were enrolled in a multicenter study and underwent surgical staging consisting of inspecting the intraperitoneal cavity, peritoneal washing, total hysterectomy, bilateral salpingo-oophorectomy, and in all cases we performed systematic bilateral pelvic lymphadenectomy by LPS or LPT approach. RESULTS: One patient of the LPS group had an uretero-vaginal fistula and another patient had an ureteral stricture temporarily treated with a stent. One patient of the LPS group had a bowel perforation due to dense adhesions with the peritoneum under the umbilicus, resolved with a bowel resection and an end-to-end anastomosis. In three patients of the LPS group we observed a vaginal cuff dehiscence and in one case a pelvic lymphocyst was reported. CONCLUSIONS: The low intraoperative and postoperative complications rate, observed in the LPS group, highlights the feasibility, safety and efficacy of this surgical approach. The operating time was longer in the LPS group but the recurrence rate and the complication rate appear similar and not more than what is traditionally expected with the LPT approach, although further studies and cost-benefit analyses are required to determine whether the use of LPS improves the outcome over standard LPT and whether the advantages of this technique could be extended to a larger proportion of patients. PMID- 20015635 TI - Inadequate pain relief and consequences in oncological elderly patients. AB - Elderly patients with cancer are particularly burdened with pain, which has an impact on physical, psychological and cognitive symptoms, and consequently, on the overall quality of life. Here, the existing literature on pain and its consequences in elderly patients with cancer is reviewed, in order to understand the impact of cancer pain and its related symptoms, and the importance of its correct assessment and management, in the geriatric population. From the literature, it emerges that cancer pain has a complex and multidimensional phenomenology in this population, and it is often underestimated and consequently untreated. Furthermore, elderly cancer patients are at higher risk of suffering from pain. Aetiology of cancer pain in elderly patients is still an emergent issue, and immunological findings on the link between pain, cancer and aging may help enlighten the pathophysiological mechanisms underlying pain in elderly cancer patients. Particularly, immune dysfunction may represent a common pathogenic ground of pain and its more common related symptoms (i.e. depression and cognitive decline) in elderly cancer patients. Appropriate pain relief represents a challenge in oncological research, in order to improve patients' and caregivers' quality of life. PMID- 20015636 TI - Starch composites reinforced by bamboo cellulosic crystals. AB - Using a method of combined HNO(3)-KClO(3) treatment and sulfuric acid hydrolysis, bamboo cellulose crystals (BCCs) were prepared and used to reinforce glycerol plasticized starch. The structure and morphology of BCCs were investigated using X-ray diffraction, electron microscopy, and solid-state (13)C NMR. Results showed that BCCs were of typical cellulose I structure, and the morphology was dependent on its concentration in the suspension. BCC of 50-100 nm were assembled into leaf nervations at low concentration (i.e. 0.1 wt.% of solids), but congregated into a micro-sized "flower" geometry at high concentration (i.e. 10.0 wt.% of solids). Tensile strength and Young's modulus of the starch/BCC composite films (SBC) were enhanced by the incorporation of the crystals due to reinforcement of BCCs and reduction of water uptake. BCCs at the optimal 8% loading level exhibited a higher reinforcing efficiency for plasticized starch plastic than any other loading level. PMID- 20015637 TI - Improvement of a two-stage fermentation process for docosahexaenoic acid production by Aurantiochytrium limacinum SR21 applying statistical experimental designs and data analysis. AB - Statistical screening experimental designs were applied to identify the significant culture variables for biomass production of Aurantiochytrium limacinum SR21 and their optimal levels were found using a combination of Artificial Neural Networks, genetic algorithms and graphical analysis. The biomass value obtained (40.3g cell dry weight l(-1)) employing the selected culture conditions agreed with that predicted by the model. Subsequently, two significant culture conditions for docosahexaenoic acid (DHA) production were determined, finding that an inoculum of 10% (v/v), obtained from the previous (statistically optimized) stage, should be used in a DHA production medium having a molar C:N ratio of 55:1, to reach a production of 7.8 g DHA l(-1) d(-1). The production step was thereafter scaled in a 3.5l bioreactor, and DHA productivity of 3.7 g l(-1) d(-1) was obtained. This two-stage strategy: statistically optimized inoculum production (fist step) and a DHA production step, is presented for the first time to optimize a bioprocess conducive to the obtention of microbial DHA. PMID- 20015638 TI - Composition and biological activity of the essential oil from leaves of Plinia cerrocampanensis, a new source of alpha-bisabolol. AB - The essential oil from fresh leaves of Plinia cerrocampanensis Barrie (Myrtaceae), obtained by hydrodistillation, was analysed by GC-FID and GC-MS. Forty components, representing more than 91% of the oil, were identified. Oxygenated sesquiterpenes represented the main fraction with alpha-bisabolol (42.8%) as the major constituent, making this plant a new and good source of this substance. Biological activity of the essential oil was evaluated against several bacterial and fungal strains as well as larvae from Aedes aegypti. The highest activity was found against Staphylococcus aureus, Pseudomonas aeruginosa, Microsporum gypseum, Trichophyton mentagrophytes and Trichophyton rubrum with MIC values from 32 to 125 microg/ml. The essential oil also showed potent inhibitory and bactericidal activities against three H. pylori strains, with MIC and MBC values of 62.5 microg/ml, and caused 100% mortality of A. aegypti larvae at a concentration of 500 microg/ml. PMID- 20015639 TI - A kinetic study of autohydrogenotrophic denitrification at the optimum pH and sodium bicarbonate dose. AB - In this study the kinetics of autohydrogenotrophic denitrification was studied under optimum solution pH and bicarbonate concentration. The optimal pH and bicarbonate concentration were firstly obtained using a design of experiment (DOE) methodology. For this purpose a total of 11 experiments were carried out. Sodium bicarbonate concentrations ranging of 20-2000 mg/L and pH values from 6.5 to 8.5 were used in the optimization runs. It was found that the pH has a more pronounced effect on the denitrification process as compared to the bicarbonate dose. The developed quadratic model predicted the optimum conditions at pH 8 and 1100 mg NaHCO(3)/L. Using these optimal conditions, the kinetics of denitrification for nitrate and nitrite degradation were investigated in separate experiments. Both processes were found to follow a zero order kinetic model. The ultimate specific degradation rates for nitrate and nitrite remediation were 29.60 mg NO(3)(-)-N/g MLVSS/L and 34.85 mg NO(3)(-)-N/g MLVSS/L respectively, when hydrogen was supplied every 0.5h. PMID- 20015640 TI - Chelating and radical scavenging activities of soy protein hydrolysates prepared from microbial proteases and their effect on meat lipid peroxidation. AB - Three commercial microbial proteases, neutral protease from Bacillus subtilis (NP), validase from Aspergillus oryze (Val), and alkaline protease from Bacillus licheniformis (AP), were investigated for producing antioxidant hydrolysates from soy protein. The resulting hydrolysates were fractioned by sequential ultrafiltration and their antioxidant properties were examined. All the 12 hydrolysate fractions showed noticeable oxygen radical absorbance capacity (ORAC) but significantly varied from 23.8 to 83.8 micromol Trolox equivalents (TE)/g. The hydrolysates also possessed significantly different 1,1-diphenyl-2 picrylhydrazyl (DPPH()) scavenging activities and transition metal chelating activities. Three fractions with strong antioxidant activities, NP-F1, Val-F1, and AP-F3, were incorporated into ground beef to determine their dose-response on lipid peroxidation during 15-day storage. AP-F3 and NP-F1 but not Val-F1 significantly reduced meat lipid peroxidation by 20.1% and 12.9%, respectively. Our results suggested that the commercial microbial proteases such as B. subtilis and B. licheniformis could be used to produce effective antioxidant hydrolysates from food proteins. PMID- 20015641 TI - Long-chain fatty acids inhibition and adaptation process in anaerobic thermophilic digestion: batch tests, microbial community structure and mathematical modelling. AB - Biomass samples taken during the continuous operation of thermophilic anaerobic digestors fed with manure and exposed to successive inhibitory pulses of long chain fatty acids (LCFA) were characterized in terms of specific metabolic activities and 16S rDNA DGGE profiling of the microbial community structure. Improvement of hydrogenotrophic and acidogenic (beta-oxidation) activity rates was detected upon successive LCFA pulses, while different inhibition effects over specific anaerobic trophic groups were observed. Bioreactor recovery capacity and biomass adaptation to LCFA inhibition were verified. Population profiles of eubacterial and archaeal 16S rDNA genes revealed that no significant shift on microbial community composition took place upon biomass exposure to LCFA. DNA sequencing of predominant DGGE bands showed close phylogenetic affinity to ribotypes characteristic from specific beta-oxidation bacterial genera (Syntrophomonas and Clostridium), while a single predominant syntrophic archaeae was related with the genus Methanosarcina. The hypothesis that biomass adaptation was fundamentally of physiological nature was tested using mathematical modelling, taking the IWA ADM1 as general model. New kinetics considering the relation between LCFA inhibitory substrate concentration and specific biomass content, as an approximation to the adsorption process, improved the model fitting and provided a better insight on the physical nature of the LCFA inhibition process. PMID- 20015642 TI - Sub-critical water hydrolysis of hog hair for amino acid production. AB - A recycling method using sub-critical water hydrolysis to convert hog hair from slaughterhouses into amino acids was developed. The influence of the reaction parameters such as temperature, time of reaction and initial substrate concentration were investigated in a batch reactor. The quality and quantity of amino acids in hydrolysates were determined and 17 kinds of amino acids were obtained. Under the tested conditions, the highest amino acid yield (325 mg/g protein) was reached at an initial substrate concentration of 10 g/l, a temperature of 250 degrees C and a reaction time of 60 min. A large amount of low molecular weight amino acids, such alanine and glycine, was observed at these operating conditions. Sub-critical water hydrolysis was confirmed as an effective and practical process to recover amino acids from hog hair waste. PMID- 20015643 TI - Dye decolorization and detoxification by laccase immobilized on porous glass beads. AB - The decolorization and detoxification of textile dyes by fungal laccase immobilized on porous glass beads were evaluated. Anthraquinone (Reactive blue 19 and Dispersed blue 3) and indigoid (Acid blue 74) dyes were degraded more rapidly than the azo dyes (Acid red 27 and Reactive black 5). There was no dye sorption to the enzyme bed when decolorization rates were high (>12 microM dye/U-h) but at moderate rates (8 to>0.06 microM/U-h), there was a transient color which disappeared upon prolonged exposure. With Reactive black 5, permanent adsorption occurred most likely because laccase had been totally inactivated. Although laccase treatment was more efficient at decolorizing the anthraquinone dyes, their toxicity (as determined by the Microtox assay) increased while the less efficiently decolorized solutions of azo and indigoid dyes became less toxic. These results demonstrate the potential and limitations of using immobilized laccase to enzymatically decolorize a range of different dye classes and reduce dye toxicity in a single step. PMID- 20015644 TI - Key issues in life cycle assessment of ethanol production from lignocellulosic biomass: Challenges and perspectives. AB - Progressive depletion of conventional fossil fuels with increasing energy consumption and greenhouse gas (GHG) emissions have led to a move towards renewable and sustainable energy sources. Lignocellulosic biomass is available in massive quantities and provides enormous potential for bioethanol production. However, to ascertain optimal biofuel strategies, it is necessary to take into account environmental impacts from cradle to grave. Life cycle assessment (LCA) techniques allow detailed analysis of material and energy fluxes on regional and global scales. This includes indirect inputs to the production process and associated wastes and emissions, and the downstream fate of products in the future. At the same time if not used properly, LCA can lead to incorrect and inappropriate actions on the part of industry and/or policy makers. This paper aims to list key issues for quantifying the use of resources and releases to the environment associated with the entire life cycle of lignocellulosic bioethanol production. PMID- 20015645 TI - Novel small molecule bradykinin B1 receptor antagonists. Part 1: benzamides and semicarbazides. AB - The synthesis and SAR of two series of bradykinin B(1) receptor antagonists is described. The benzamide moiety proved to be a suitable replacement for the aryl ester functionality of biaryl based antagonists. In addition, it was found that semicarbazides can effectively replace cyclopropyl amino acids. The compounds with the best overall profile were biaryl semicarbazides which display high antagonistic activity, low Caco-2 efflux and high oral bioavailability in the rat. PMID- 20015646 TI - Inhibition of Yersinia protein tyrosine phosphatase by phosphonate derivatives of calixarenes. AB - Inhibition of Yersinia protein tyrosine phosphatase by calix[4]arene mono-, bis-, and tetrakis(methylenebisphosphonic) acids as well as calix[4]arene and thiacalix[4]arene tetrakis(methylphosphonic) acids have been investigated. The kinetic studies revealed that some compounds in this class are potent competitive inhibitors of Yersinia PTP with inhibition constants in the low micromolar range. The binding modes of macrocyclic phosphonate derivatives in the enzyme active center have been explained using computational docking approach. The results obtained indicate that calix[4]arenes are promising scaffolds for the development of inhibitors of Yersinia PTP. PMID- 20015647 TI - Conversion of 4-cyanomethyl-pyrazole-3-carboxamides into CB1 antagonists with lowered propensity to pass the blood-brain-barrier. AB - A series of amides, amidines and amidoximes have been made from the corresponding nitrile compounds, to provide potent antagonists and inverse agonists for the CB1 receptor with considerably lower lipophiliciy, higher polar surface area and improved plasma/brain ratios compared to the centrally acting rimonabant. Extensive investigations of ADME and in vivo pharmacological properties led to selection of the amide series and specifically the 4-(4-fluorophenyl)piperidin-4 ol derivative D4. A clear improvement in the peripheral profile over rimonabant was seen, although some contribution of central effect on the pronounced weight reduction in obese mice cannot be ruled out. PMID- 20015648 TI - Discovery and SAR of novel, potent and selective hexahydrobenzonaphthyridinone inhibitors of poly(ADP-ribose)polymerase-1 (PARP-1). AB - A novel hexahydrobenzonaphthyridinone PARP-1 pharmacophore is reported, subsequent SAR exploration around this scaffold led to selective PARP-1 inhibitors with low nanomolar enzyme potency, displaying good cellular activity and promising rat PK properties. PMID- 20015649 TI - Biaryl piperidines as potent and selective delta opioid receptor ligands. AB - The design and synthesis of novel opiates are reported. Based on the message address principle a novel class of 4,4- and 3,3-biaryl piperidines was designed and synthesized. Biological evaluation confirmed that these compounds exhibit high affinity and selectivity for the delta opioid receptor. Key structure activity relationships that influence affinity, selectivity, functional activity and clearance are reported. PMID- 20015650 TI - Protection of a single-cysteine redox switch from oxidative destruction: On the functional role of sulfenyl amide formation in the redox-regulated enzyme PTP1B. AB - Model reactions offer a chemical mechanism by which formation of a sulfenyl amide residue at the active site of the redox-regulated protein tyrosine phosphatase PTP1B protects the cysteine redox switch in this enzyme against irreversible oxidative destruction. The results suggest that 'overoxidation' of the sulfenyl amide redox switch to the sulfinyl amide in proteins is a chemically reversible event, because the sulfinyl amide can be easily returned to the native cysteine thiol residue via reactions with cellular thiols. PMID- 20015651 TI - Novel small molecule bradykinin B1 receptor antagonists. Part 2: 5-membered diaminoheterocycles. AB - Efforts to find new bradykinin B(1) receptor antagonists identified 2 aminobenzimidazole as a novel core. Subsequent transformation into five-membered diaminoheterocycle derivatives and their synthesis and SAR is described. This resulted in compounds with low nanomolar activity. PMID- 20015652 TI - Breast cancer with non-inflammatory skin involvement: current data on an underreported entity and its problematic classification. AB - We evaluated 166 breast cancer cases with non-inflammatory skin involvement (NISI), which were classified in the TNM classification as T4b. The distribution of tumour sizes and stages was: < or =3 cm:24.1%, 3.1-5 cm:21.7%, 5.1-10 cm:33.1%, >10 cm:21.1%; stages:I/II:21.0%, III:43.4%, IV:35.6%. To assess the impact of NISI on axillary lymph node involvement (ALNI), we analyzed a sub-group of 50 patients with tumours < or =5 cm and compared them with a matched control group. NISI was found to be associated with increased ALNI (HR, 2.66; 95%CI, 1.59 4.63; p<0.0001). According to the inherent rules of tumour classification, only tumours with similar morphologic extent and prognostic significance should be combined. Since there is a high grade of heterogeneity, this basic tenet is clearly violated regarding breast cancer with NISI. Our proposal is to eliminate these tumours from the T4 category and to classify them simply by size (T1-3). Due to its prognostic significance, NISI should be indicated by an optional descriptor (e.g. S1). PMID- 20015653 TI - RNA secondary structural determinants of miRNA precursor processing in Arabidopsis. AB - MicroRNAs (miRNAs) are excised from hairpin structures within primary miRNAs (pri miRNAs). Most animal pri-miRNAs are processed by two cleavages, the first at a loop-distal site approximately 11 nucleotides (nt) from the end of the hairpin and the second approximately 22 nt beyond the first. To identify RNA structural determinants of miRNA processing in plants, we analyzed the functional consequences of changing the secondary structure of the lower (loop-distal), middle (miRNA:miRNA(*)), and upper (loop-proximal) stems of the hairpin in two different pri-miRNAs. Closing bulges immediately below the loop-distal cleavage sites increased the accumulation of accurately cleaved precursor miRNAs but decreased the abundance of the mature miRNAs. A pri-miRNA variant with an unpaired lower stem was not processed, and variants with a perfectly paired middle or upper stem were processed normally. Bioinformatic analysis of pri-miRNA structures, together with physical mapping of initial cleavage sites and in vitro processing of pri-miRNA, reveals that the first, loop-distal cleavage is often at a distance of approximately 15 nt from an unpaired region. Hence, a common determinant of the rate and location of the initial pri-miRNA cleavage is an imperfectly base-paired duplex of approximately 15 nt between the miRNA:miRNA(*) duplex and either a less structured region of the lower stem or its end. PMID- 20015654 TI - Structure determinants for accurate processing of miR172a in Arabidopsis thaliana. AB - Plant microRNAs (miRNAs) are processed by the RNase III-like enzyme DICER-LIKE1 acting in concert with the double-stranded RNA-binding protein HYPONASTIC LEAVES1 and the zinc finger protein SERRATE. Together, they excise a miRNA/miRNA( *) duplex with a 2 nucleotide 3' overhang from the primary miRNA (pri-miRNA) transcript. pri-miRNAs include a partially self-complementary foldback or stem loop, which gives rise to the mature miRNA. In animals, pri-miRNAs are very similar, with a stereotypic position of the miRNA within the foldback. Accordingly, rules for miRNA excision from the precursor are quite simple in animals. In contrast, how miRNA sequences are recognized in the structurally much more diverse foldbacks of plants is unknown. We have performed an extensive in vivo structure-function analysis of Arabidopsis thaliana pri-miRNA 172a (pri miR172a). A junction of single-stranded and double-stranded RNA 15 nucleotides proximal from the miRNA/miRNA(*) duplex appears to be essential for accurate miR172a processing. This attribute is found in several other but not all plant miRNA foldbacks. In addition, we have identified features of the distal foldback structure important for miR172a processing. Our ability to engineer de novo a functional minimal miRNA precursor highlights that we have discovered several elements both necessary and sufficient for accurate miRNA processing. PMID- 20015655 TI - Phase retrieval low energy electron microscopy. AB - We consider the utility of phase-retrieval methods in low energy electron microscopy (LEEM). Computer simulations are presented, demonstrating recovery of the terraced height profile of atomic steps. This recovery uses phase retrieval to decode a single LEEM image, incorporating the effects of defocus, spherical aberration and chromatic aberration. The ability of the method, to obtain temporal sequences of evolving step profiles from a single LEEM movie, is discussed. PMID- 20015656 TI - Elemental analysis of two Egyptian phosphate rock mines by instrumental neutron activation analysis and atomic absorption spectrometry. AB - Because of the increasing use of phosphate in industry world wide, it is interesting to investigate the elemental concentration in phosphate ores. The present work determines the elemental analysis of two different types of phosphate ores, containing different amounts of P(2)O(5) to check the level of the radioactive elements U and Th and of stable environmental pollutants like Cr, Zn in phosphate raw material. In addition, rare earth and other elements are analysed by the INAA method. This knowledge is found to be important in the assessment of possible radiological hazards to human health, since these materials may be used as building material or as phosphatic fertilizers and animal feed ingredients. PMID- 20015657 TI - Discordant findings in patients with non-small-cell lung cancer: absolutely normal bone scans versus disseminated bone metastases on positron-emission tomography/computed tomography. AB - OBJECTIVE: At present, metastatic bone involvement is usually assessed using bone scintigraphy, which has a high sensitivity but a poor specificity. The objective of our study was to compare the sensibility of the 2-deoxy-2-[18F] fluoro-d glucose positron emission tomography/computed tomography (F-18 FDG PET/CT) for the detection of bone metastasis in patients with non-small-cell lung cancer (NSCLC) whose technetium 99m methylenediphosphonate (Tc-99m MDP) bone scans were absolutely normal. MATERIAL AND METHODS: This study based on the retrospective analysis of 95 consecutive patients with histologically proven NSCLC who underwent F-18 FDG PET/CT and Tc-99m MDP bone scan at the Eskisehir Osmangazi University School of Medicine, Department of Nuclear Medicine between November 2006 and October 2008. Nineteen patients (19 of 95, 20%) with absolutely normal Tc-99m bone scan versus multiple high-grade F-18 FDG avid bony metastases on F-18 FDG PET/CT were selected for the review. Their ages ranged from 46 to 73 years (15 males and four females; mean: 57.2 years). RESULTS: Nine patients had squamous cell carcinoma, six had adenocarcinoma, three had large cell carcinoma and one had adenosquamous cell carcinoma. Tc-99m MDP bone scan that did not reveal bony abnormalities or radiotracer uptake was characteristic of benign disease (defined as absolutely normal) in these patients. Whereas, F-18 FDG PET/CT not only showed extremely disseminated heterogeneous nest-like high-grade FDG avid metastatic foci within the marrow cavity of the upper and lower thoracic spine, lumbar spine, pelvis, rib cages and bilateral proximal long bones, but also showed disseminated osteolytic bony metastases in these areas. CONCLUSION: Discordant findings of skeletal metastasis between Tc-99m MDP bone scans and F-18 FDG PET/CT imaging may be seen in 20% of the patients with NSCLC. F-18 FDG PET/CT could detect metastatic bone involvement more accurately than bone scintigraphy. Bone scans are insensitive to early bone marrow neoplastic infiltration. Assessment of glucose metabolism with FDG PET/CT can represent a more powerful tool to detect early bone metastases in lung cancer than with traditional bone scans. PMID- 20015658 TI - Aortic valve insufficiency with extensive cusp pathology: repair with a pericardial patch. AB - The novel repair technique for aortic insufficiency with extensive cusp pathology using autologous pericardium is described. The technique has been used in 19 consecutive patients offering excellent functional results with a large coaptation area. PMID- 20015659 TI - Expansion of beta-cell mass in response to pregnancy. AB - Inadequate beta-cell mass can lead to insulin insufficiency and diabetes. During times of prolonged metabolic demand for insulin, the endocrine pancreas can respond by increasing beta-cell mass, both by increasing cell size and by changing the balance between beta-cell proliferation and apoptosis. In this paper, we review recent advances in our understanding of the mechanisms that control the adaptive expansion of beta-cell mass, focusing on the islet's response to pregnancy, a physiological state of insulin resistance. Functional characterization of factors controlling both beta-cell proliferation and survival might not only lead to the development of successful therapeutic strategies to enhance the response of the beta-cell to increased metabolic loads, but also improve islet transplantation regimens. PMID- 20015661 TI - Local serum levels of vascular endothelial growth factor in infantile hemangioma: intriguing mechanism of endothelial growth. AB - The pathogenesis of hemangiomas still remains poorly understood. Dysregulation of angiogenesis has been proposed to play a central role in hemangioma pathogenesis. The aim of our study was to determine the peripheral and local serum levels of VEGF in patients with hemangiomas and vascular malformations. MATERIAL AND METHODS: The study group consisted of 52 children with infantile hemangioma (33 with proliferative lesions, 19 with involuting lesions), 14 children with vascular malformations and 36 healthy children. VEGF serum levels were analyzed by an ELISA assay and the values between the groups were compared. RESULTS: The serum peripheral VEGF concentrations in children with proliferative hemangiomas were significantly higher than in patients with involuting hemangiomas, vascular malformations and controls. There was no correlation between the measured cytokine level, hemangioma size, and the age of the patients. The local serum VEGF levels in 29 children with hemangiomas were distinctly lower than in the peripheral blood, both in 20 proliferating hemangiomas (p<0.0001) and 9 involuting ones (p=0.007); and the difference between females and males was non significant (NS p=0.06). CONCLUSIONS: (1) VEGF serum levels vary in the different phases of hemangioma growth and may help to distinguish hemangiomas from vascular malformations; (2) obtained local results may support the intrinsic theory of endothelial cell proliferation in hemangiomas. PMID- 20015660 TI - Thyroid hormone crosstalk with nuclear receptor signaling in metabolic regulation. AB - Thyroid hormone influences diverse metabolic pathways important in lipid and glucose metabolism, lipolysis and regulation of body weight. Recently, it has been recognized that thyroid hormone receptor interacts with transcription factors that predominantly respond to nutrient signals including the peroxisome proliferator-activated receptors, liver X receptor and others. Crosstalk between thyroid hormone signaling and these nutrient responsive factors occurs through a variety of mechanisms: competition for retinoid X receptor heterodimer partners, DNA binding sites and transcriptional cofactors. This review focuses on the mechanisms of interaction of thyroid hormone signaling with other metabolic pathways and the importance of understanding these interactions to develop therapeutic agents for treatment of metabolic disorders, such as dyslipidemias, obesity and diabetes. PMID- 20015662 TI - Transcriptional regulation of cytokines and oxidative stress by gallic acid in human THP-1 monocytes. AB - Increased inflammation/prooxidation has been linked not only to Type 2 diabetes but also in prediabetes state. In this study we investigated hyperglycemia mediated proinflammatory/prooxidant effects in THP-1 monocytes and tested whether gallic acid could attenuate changes in gene expression induced by high-glucose. Cells were treated either with 5.5mM glucose or 25mM glucose in the absence and presence of gallic acid. While oxidative DNA damage was assessed by COMET assay, GSH and GSSG levels were estimated fluorimetrically. Gene expression patterns were determined by RT-PCR. Cells treated with high-glucose showed increased DNA damage and glutathione depletion and this was attenuated in the presence of gallic acid. High-glucose treated cells exhibited increased mRNA expression of TNF-alpha, IL-6, NADPH oxidase and TXNIP and gallic acid attenuated these proinflammatory and prooxidant effects. Cells treated with high-glucose revealed a deficiency in mounting SOCS-3 expression and gallic acid upregulates this feedback regulatory signal. Gallic acid attenuates DNA damage, maintains glutathione turnover, and suppresses hyperglycemia-induced activation of proinflammatory and prooxidant gene expression. Gallic acid beneficially modulate transcription of functionally diverse groups of genes and its regulation of SOCS 3 and TXNIP signals is a newly identified mechanism that has therapeutic implications. PMID- 20015663 TI - Roles of endothelin-1 and selected proinflammatory cytokines in the pathogenesis of proliferative diabetic retinopathy: Analysis of vitreous samples. AB - PURPOSE: To investigate the roles of endothelin-1(ET-1), TNF-alpha, IL-6 in the pathogenesis of proliferative diabetic retinopathy (PDR) in type 2 diabetes. METHODS: Vitreous and blood serum samples were collected during vitrectomy from 19 patients with PDR and 15 patients who underwent vitrectomy for other reasons. The concentrations of ET-1, TNF-alpha, IL-6, vWF, sE-selectin were determined by ELISA. RESULTS: Intraocular and serous concentrations of ET-1, TNF-alpha, IL-6, vWF, sE-selectin were higher in patients with PDR than in the control group. The vitreous ET-1/plasma ET-1 ratios the group of diabetic patients and in the control group were similar. Also TNF-alpha, IL-6 vitreous/plasma ratio were not statistically different between the analysed groups. Correlation between intraocular ET-1 and TNF-alpha concentrations in patients with PDR and between the increases in both factors in the vitreous and HbA(1)c concentration were shown. In the vitreous the increase in vWF depended on elevated levels of vWF in the serum. E-selectin concentration correlated with diastolic blood pressure. CONCLUSION: These data provide evidence of the activation of the local synthesis of ET-1, TNF-alpha, IL-6 in PDR. The relationship between the increase in vitreous ET-1, TNF-alpha concentrations and HbA(1)c concentration is a important confirmation of the necessity to optimise diabetes treatment. PMID- 20015664 TI - On the nature of cognitive control and endogenous orienting: a response to Chica and Bartolomeo (2010). AB - Chica and Bartolemeo (Unconscious strategies? Commentary on Risko and Stolz (2010): The proportion valid effect in covert orienting: Strategic control or implicit learning? Consciousness and Cognition,19, 443-444.) agree that our results (Risko, E. F., & Stolz, J. A. (2010). The proportion valid effect in covert orienting: Strategic control or implicit learning? Consciousness and Cognition,19, 432-442.) are consistent with an implicit learning account of the proportion valid effect. Nevertheless, they raise two general issues (1) that an explicit strategy might be operative in other contexts and (2) that orienting in response to implicit knowledge is endogenous. In our response, we address each of these issues and further discuss the concepts of endogenous orienting and cognitive control. PMID- 20015665 TI - Treatment of metastatic prostate adenocarcinoma to the calcaneus. AB - Metastatic skeletal adenocarcinoma is an all too common and unfortunate complication of advanced oncologic states. Mortality rates are usually elevated when bony metastasis are evident, as this signifies advanced disease. The foot and ankle are uncommon sites for metastatic deposits, but may occur. As such, the foot and ankle surgeon must be aware of the potential for such disease, and be able to proceed with an imaging and medical work-up of the patient with foot and ankle skeletal metastasis. The goal of treatment is pain relief and the preservation of functional ambulation, which may greatly enhance the quality of remaining life for patients. A team approach is mandatory to manage the patients with metastatic disease. We present a case of an elderly male with a known history of prostate cancer, who presented with unrelenting heel pain, which upon diagnostic work-up, proved to be progressive calcaneal as well as axial metastasis after a brief period of clinical remission. Operative management coupled with palliative radiation and bisphosphonate therapy provided symptomatic pain relief and maintenance of functional ambulation. PMID- 20015666 TI - A review of 51 talonavicular joint arthrodeses for flexible pes valgus deformity. AB - The purpose of this study was to evaluate the radiographic and clinical outcomes of isolated talonavicular arthrodesis in the treatment of the flexible pes valgus foot type. Retrospectively, 51 consecutive isolated talonavicular arthrodeses in 41 patients were evaluated. The mean patient age was 47 (range 9 to 72) years, and the mean follow-up duration was 43.3 (range 11 to 113) months. The mean 10-cm categorical pain score before the surgery was 7.60 +/- 2.37, and this improved to 1.90 +/- 2.38 postoperatively, and this difference was statistically significant (P < .001). We also measured the preoperative and postoperative percent of talar uncovering, calcaneocuboid angle, Kite's angle, talar dome height, calcaneal inclination angle, and Meary's angle, and observed statistically significant (P < .001) improvements in all of these. Fifty-one (100%) of the cases progressed to radiographic osseous union, although 2 (3.92%) of the cases were considered delayed unions. Moreover, 4 (7.84%) of the cases displayed juxta-articular arthrosis postoperatively, and 2 (3.9%) cases developed transfer pain to the lateral column. The authors concluded that isolated talonavicular arthrodesis is a safe and effective procedure for the treatment of painful pes valgus deformity. PMID- 20015667 TI - Reimplantation of a totally extruded talus: a case report. AB - Total extrusion of the talus is an unusual injury, and the obvious risks of reimplantation of the extruded bone include infection and avascular necrosis. In this article, the authors present the case of a 34-year-old man who sustained an open ankle injury with complete extrusion of the talus. The talus was recovered at the scene of the accident, and subsequently reimplanted along with ankle stabilization with pins and an external fixator. At 6 weeks following the osseous surgery, final soft tissue reconstruction with a suralis flap was performed. At 3 years after the injury, radiographs revealed spontaneous fusion of the tibiotalar and subtalar joints, and the clinical examination and history indicated satisfactory weight-bearing function of the involved foot and ankle. The definitive treatment of this serious lower extremity injury remains controversial, and the use of large allogeneic bone grafts, vascularized bone grafts, and tibiocalcaneal fusion, as well as reimplantation of the extruded talus have been recommended. PMID- 20015668 TI - The positive effect of immunosuppression on adaptation of venous allografts to arterialisation in rats. AB - OBJECTIVES AND DESIGN: We investigated whether immunosuppression was necessary for transplanted allogeneic veins to adapt to arterialisation. We used a transplant rat model with or without immunosuppression. MATERIAL AND METHODS: Iliolumbar veins from Lewis (LEW) or Brown-Norway (BN) rats were transplanted into the abdominal aorta of isogeneic (LEW to LEW; group A) or allogeneic (BN to LEW; groups B and C) rats. Group C had daily intramuscular injections of 0.2mgkg( 1) FK506. Light microscope evaluations of grafts were performed at 30 days following transplantation. We determined the presence of endothelial cells, the intensity of intimal proliferation and the degree of infiltration by Lewis major histocompatibility complex (MHC) class II positive, CD4-positive and CD8-positive cells into the adventitia. RESULTS: Groups A and C displayed similar results in intimal thickness (12.7+/-7.0microm vs. 15.0+/-8.4 mum, respectively) and degree of adventitial infiltration by MHC class II positive (16.6+/-7.5 vs. 14.6+/-6.2, respectively), CD8-positive (0.8+/-1.7 vs. 1.8+/-2.6, respectively) and CD4 positive (12.5+/-7.7 vs. 5.8+/-4.6, respectively) cells. In contrast, allogeneic rats without immunosuppression (group B) showed infiltration of host immunocompetent cells and destruction of the venous wall with no histological signs of arterialisation. CONCLUSION: Immunosuppressive therapy is necessary for venous allograft adaptation to arterialisation in rats. PMID- 20015669 TI - Tail biting: an important and undesirable behaviour of growing pigs. PMID- 20015670 TI - Movement disorders in neuro-metabolic diseases. AB - Inborn errors of metabolism (IEM) are a group of genetic disorders characterized by dysfunction of an enzyme or other protein involved in cellular metabolism.(1) Most IEMs involve the nervous system (neuro-metabolic diseases or NMD). NMD often present with a complex clinical picture: psychomotor retardation and/or regression, pyramidal signs, ataxia, hypotonia and epilepsy and movement disorders.(1) Movement disorders are more frequently part of this complex picture than a predominant symptom, however in some instances the clinical picture may be summarized in an invalidating movement disorder.(2) On a phenomenology basis, one can distinguish eight main types of movement disorders: dystonia and athetosis, chorea, tremor with or without parkinsonism, ballismus, myoclonus, tics and stereotypies. Most of these abnormal involuntary movements generate from a dysfunction or a lesion in the basal ganglia, excepting myoclonus, the origin of which can vary (cortical, brainstem, basal ganglia, spinal and even peripheral nervous system).(3) Classically the most frequently observed movement disorders in NMD are: dystonia, myoclonus, chorea, tremor and parkinsonism (Fig. 1). The primary goal of this article is, departing from the literature and a large personal series, to describe the types of movement disorders most frequently observed in NMD and to discuss their clinical value in the setting of specific types of NMD. PMID- 20015671 TI - DANGLE: A Bayesian inferential method for predicting protein backbone dihedral angles and secondary structure. AB - This paper introduces DANGLE, a new algorithm that employs Bayesian inference to estimate the likelihood of all possible values of the backbone dihedral angles phi and psi for each residue in a query protein, based on observed chemical shifts and the conformational preferences of each amino acid type. The method provides robust estimates of phi and psi within realistic boundary ranges, an indication of the degeneracy in the relationship between shift measurements and conformation at each site, and faithful secondary structure state assignments. When a simple degeneracy-based filtering procedure is applied, DANGLE offers an ideal compromise between accuracy and coverage when compared with other shift based dihedral angle prediction methods. In addition, per residue analysis of shift/structure degeneracy has potential to be a useful new approach for studying the properties of unfolded proteins, with sufficient sensitivity to identify regions of residual structure in the acid denatured state of apomyoglobin. PMID- 20015672 TI - Dependence of sonochemical parameters on the platinization of rutile titania - an observation of a pronounced increase in photocatalytic efficiencies. AB - Using a standing wave sonochemical reactor (SWSR), the influences of parameters of ultrasonic power input, sonication time, sonication temperature and the amount of propanol (which generates the reducing radicals) were systemically investigated to ascertain and optimize the best conditions for the sonochemical reduction of Pt from its precursor hexachloroplatinic acid and then its deposition on rutile TiO(2) (platinization of rutile titania) catalysts. Catalytic activity of the prepared platinized catalysts was tested in the reaction of methyl orange degradation. The results of photocatalytic activity study in the degradation of methyl orange further demonstrate that sonochemically as-prepared Pt/TiO(2) catalysts show a pronounced increase ( approximately 2 times) in photodegradation, even with a deposition of small amounts of platinum (1.4wt.%), as compared to the unsupported or naked rutile titania. Although there are various parameters that influence the sonochemical platinization of rutile titania, the present optimization results clearly indicate that the best photocatalytic degradation of methyl orange can be obtained when the experimental conditions of the preparation were with an input power of 50W, an initial hexachloroplatinic acid volume of 70ml (which results into 1.4wt.% Pt on TiO(2)), sonication time of 90min, 0.18g of propanol and a temperature of 10 degrees C were adopted. The method of ultrasound application to prepare metal supported semiconductors has many advantages such as convenience, safety and high efficiency. Furthermore, it is hopeful that this optimization study can also be extended to the generation of similar metal supported semiconductors. PMID- 20015673 TI - Effect of ultrasound processing on anthocyanins and color of red grape juice. AB - Grape juice samples were sonicated with processing variables of amplitude level (24.4-61.0microm) and treatment time (0-10min) at a constant frequency of 20kHz and pulse durations of 5s on and 5s off. A full factorial experimental design with regression modeling was employed to investigate the main effects of amplitude level and treatment time on anthocyanins and color parameters. Significant effects of sonication on major anthocyanins cyanidin-3-O-glucosides (CA), malvanidin-3-O-glucosides (MA) and delphinidin-3-O-glucosides (DA), color values (L*, a*, b*) and color index (CI) were observed. Prediction models were found to be significant (p<0.05) with low standard errors and high coefficients of determination (R(2)). Model predictions for critical quality parameters of anthocyanins (CA; MA; DA), color values (L*, a*, b*), TCD and CI inactivation were closely correlated to the experimental results obtained. Significant retention of anthocyanin content in grape juice was observed for CA (97.5 %); MA (48.2 %) and DA (80.9%) during sonication. CI and other color combinations (L*a*b*, L*a*/b* and L*b*/a*) were found to be strongly correlated with anthocyanin content. This study shows that sonication could be employed for as a preservation technique for fruit juice processing where anthocyanin retention is desired. PMID- 20015674 TI - Auditory evoked potentials for monitoring during anaesthesia: a study of data quality. AB - The auditory evoked potential termed the middle latency response (MLR) has been suggested as an indicator of adequacy of anaesthesia during surgery. However, the response is small and must be extracted from high levels of background noise. A key consideration in using the MLR for clinical monitoring is whether data quality is sufficient to detect small changes. The aim of this study was to investigate the quality of the MLR recorded during anaesthesia, as a rigorous analysis of data quality is lacking in many studies. MLR recordings from patients sedated in intensive care after cardiac surgery were compared to recordings from a reference group of young volunteers with normal hearing. Data quality was measured with the F(sp) parameter. A bootstrap analysis was used to measure statistical response presence and to detect within-subject changes during clinical anaesthesia. Noise levels were high in the normative group probably due to myogenic and EEG activity. With 5 Hz click stimulation, MLR presence in the normative group was below 30%. Response presence improved using stimulation paradigms with chirps or maximum length sequences and reached 100% with a combination of maximum length sequences and chirps. F(sp) values generally improved during anaesthesia as noise levels reduced and MLR presence was 100% for MLS click stimulation. Changes in the MLR amplitude with propofol infusion rate were small. Some within-subject changes in MLR amplitude were detected using the bootstrap analysis, but 100% detection was not possible. CONCLUSION: Obtaining good quality MLR data in awake subjects is challenging. Data quality improves during clinical anaesthesia and with advanced stimulation methods, but reliable detection of changes in the MLR for clinical monitoring remains a challenge. PMID- 20015675 TI - An automatic geometrical and statistical method to detect acoustic shadows in intraoperative ultrasound brain images. AB - In ultrasound images, acoustic shadows appear as regions of low signal intensity linked to boundaries with very high acoustic impedance differences. Acoustic shadows can be viewed either as informative features to detect lesions or calcifications, or as damageable artifacts for image processing tasks such as segmentation, registration or 3D reconstruction. In both cases, the detection of these acoustic shadows is useful. This paper proposes a new method to detect these shadows that combines a geometrical approach to estimate the B-scans shape, followed by a statistical test based on a dedicated modeling of ultrasound image statistics. Results demonstrate that the combined geometrical-statistical technique is more robust and yields better results than the previous statistical technique. Integration of regularization over time further improves robustness. Application of the procedure results in (1) improved 3D reconstructions with fewer artifacts, and (2) reduced mean registration error of tracked intraoperative brain ultrasound images. PMID- 20015676 TI - Estimating zero-strain states of very soft tissue under gravity loading using digital image correlation. AB - This paper presents several experimental techniques and concepts in the process of measuring mechanical properties of very soft tissue in an ex vivo tensile test. Gravitational body force on very soft tissue causes pre-compression and results in a non-uniform initial deformation. The global digital image correlation technique is used to measure the full-field deformation behavior of liver tissue in uniaxial tension testing. A maximum stretching band is observed in the incremental strain field when a region of tissue passes from compression and enters a state of tension. A new method for estimating the zero-strain state is proposed: the zero strain position is close to, but ahead of the position of the maximum stretching band, or in other words, the tangent of a nominal stress stretch curve reaches minimum at lambda greater or similar 1. The approach, to identify zero strain by using maximum incremental strain, can be implemented in other types of image-based soft tissue analysis. The experimental results of 10 samples from seven porcine livers are presented and material parameters for the Ogden model fit are obtained. The finite element simulation based on the fitted model confirms the effect of gravity on the deformation of very soft tissue and validates our approach. PMID- 20015677 TI - What should be compared in comparative mental time travel? PMID- 20015678 TI - Nutritional immunity beyond iron: a role for manganese and zinc. AB - Vertebrates sequester iron from invading pathogens, and conversely, pathogens express a variety of factors to steal iron from the host. Recent work has demonstrated that in addition to iron, vertebrates sequester zinc and manganese both intracellularly and extracellularly to protect against infection. Intracellularly, vertebrates utilize the ZIP/ZnT families of transporters to manipulate zinc levels, as well as Nramp1 to manipulate manganese levels. Extracellularly, the S100 protein calprotectin sequesters manganese and potentially zinc to inhibit microbial growth. To circumvent these defenses, bacteria possess high affinity transporters to import specific nutrient metals. Limiting the availability of zinc and manganese as a mechanism to defend against infection expands the spectrum of nutritional immunity and further establishes metal sequestration as a key defense against microbial invaders. PMID- 20015679 TI - Baeyer-Villiger monooxygenases: recent advances and future challenges. AB - Baeyer-Villiger monooxygenases For many enzyme classes, a wealth of information on, for example, structure and mechanism has been generated in the last few decades. While the first Baeyer-Villiger monooxygenases (BVMOs) were already isolated more than 30 years ago, detailed data on these enzymes were lacking until recently. Over the last years several major scientific breakthroughs, including the elucidation of BVMO crystal structures and the identification of numerous novel BVMOs, have boosted the research on BVMOs. This has led to intensified biocatalytic explorations of novel BVMOs and structure-inspired enzyme redesign. This review provides an overview on the recently gained knowledge on BVMOs and sketches the outlook for future industrial applications of these unique oxidative biocatalysts. PMID- 20015680 TI - Type III secretion systems shape up as they ship out. AB - Virulence associated protein type III secretion systems (T3SSs) are intricately structured organic nanosyringes that achieve the translocation of bacterial proteins from the prokaryotic cytoplasm across three membranes into the host cytosol. The substrates for these systems number in the hundreds, with remarkably diverse biological activities, modulating host cell biology for the benefit of the pathogen. Although there has been tremendous progress on the structure and function of the T3SS substrates, there has been comparatively little progress on the much more highly conserved secretion apparatus itself. This review summarizes recent advances in the field of structural microbiology that have begun to address this shortcoming, finally bringing to bear the power of structural biology to this central virulence system of Gram-negative bacterial pathogens. PMID- 20015681 TI - Experimental and theoretical study of a new carbazole derivative having terminal benzimidazole rings. AB - A novel intramolecular donor-acceptor compound has been synthesized and characterized. This compound was a symmetrical A-pi-D-pi-A type molecule containing two benzimidazole rings as two electron acceptors (A) and an N ethylcarbazole group as electron donors (D). The absorption and emission spectra of the compound were determined by experimental methods in solution and were computed by using the density functional theory (DFT) and the time-dependent density functional theory (TDDFT) in gas phase and in chloroform solution. The calculated absorption and emission wavelengths were in good agreement with the experimental ones. The fluorescence quantum yields and fluorescence lifetimes of the compound in several solvents have been studied by means of steady state and time resolved fluorescence. The results showed the compound had high quantum yield. The cross-section of two-photon absorption (TPA) of the compound was measured by using femtosecond laser in dichloromethane solution. The result indicated the cross-section maximum of two-photon absorption was 430 GM at 600 nm. These results made the compound of great interest as a new fluorescent probe and photoluminescence material. PMID- 20015682 TI - Study on fluorometric discrimination of phytoplankton based on time-series vectors of wavelet transform. AB - The feasibility of using time domain of wavelet transform as characteristics to establish a fluorometric discrimination method of phytoplankton was discussed. Twelve phytoplankton species belonging to nine genera of five divisions were studied. Five steps were introduced: firstly, the feasibility of utilizing 3D fluorescence spectra (3D-FS) to discriminate phytoplankton was discussed; the relative standard deviation (RSD) and included angle cosine (IAC) were used as the test criterion. 3D-FS had such potentials, for most RSD were <5% and most IAC were >0.990. Secondly, the 3D-FS were decomposed by db7 wavelet and time-series vectors (TSVs) were generated. Thirdly, the optimal characteristic spectra (OCS) were selected from the TSV by Bayesian linear discriminant analysis (BLDA). The ability of OCS to classify phytoplankton was tested, and the correct classification ratios (CCRs) at different levels were obtained. Most CCRs were 90 100% at the species level. They were >98% at the genus level, and >99% at the division level. Fourthly, the growth and light stability of the OCS were tested. Both stabilities were high with lower RSD (<3%) and higher IAC (>0.999) compared with 3D-FS. Fifthly, a "database of reference spectra" consisting of 46 reference spectra was established by hierarchical cluster analysis (HCA). Based on this, the discrimination method of phytoplankton species was established by nonnegative least squares (NNLSs). Most reference spectra were representative to phytoplankton species; and had moderate anti-noise ability: With noise 98% at the genus level and >99% at the division level. 20% noise was a larger interference which made CDRs down to 85% at the genus level and to 99% at the division level. A fluorometric discrimination method of phytoplankton could be established based on TSV of wavelet transform. PMID- 20015683 TI - Nonradiative inter- and intramolecular energy transfer from the aromatic donor anisole to a synthesized photoswitchable acceptor system. AB - We report steady state and time resolved fluorescence measurements on acetonitrile (ACN) solutions of the model compounds, energy donor anisole (A) and a photoswitchable acceptor N,N'-1,2-phenylene di-p-tosylamide (B) and the multichromophore (M) where A and B are connected by a spacer containing both rigid triple (acetylenic) and flexible methylene bonds. Both steady state and time correlated single photon counting measurements demonstrate that though intermolecular energy transfer, of Forster type, between the donor and acceptor moieties occurs with rate 10(8)s(-1) but when these two reacting components are linked by a spacer (multichromophore, M) the observed transfer rate ( approximately 10(11)s(-1)) enhances. This seemingly indicates that the imposition of the spacer by inserting a triple bond may facilitate in the propagation of electronic excitation energy through bond. The time resolved fluorescence measurements along with the theoretical predictions using Configuration interaction singles (CIS) method by using 6-31G (d,p) basis set, implemented in the Gaussian package indicate the formations of the two excited conformers of B. The experimental findings made from the steady state and time resolved fluorescence measurements demonstrate that, though two different isomeric species of the acceptor B are formed in the excited singlet states, the prevailing singlet-singlet nonradiative energy transfer route was found from the donor A to the relatively longer-lived isomeric species of B. PMID- 20015684 TI - Development and performance of a new recombinant virus phenotypic entry assay to determine HIV-1 coreceptor usage. AB - BACKGROUND: Clinical trials of CCR5 antagonists have relied on the phenotypic determination of HIV-1 coreceptor usage. Few phenotypic assays are available, with few data on their concordance, and none has been designed to determine tropism from cell-associated HIV-1 DNA. OBJECTIVES: To assess the performance of the new Toulouse Tropism Test (TTT) phenotypic assay to characterize HIV-1 tropism using blood plasma and peripheral blood mononuclear cells (PBMC). STUDY DESIGN: 434 plasma and 168 PBMC samples were tested with the TTT assay. We determined the correlation between our assay results on plasma samples and those of the commercial Trofile assay. RESULTS: The TTT assay determined the tropism of 97% of samples after successful amplification of the env gene. It performed well on both cell samples and plasma samples with various HIV-1 loads and subtypes. It detected 0.5% of minor CXCR4-using variants in the virus population. The TTT and the Trofile assays were >90% concordant for predicting HIV-1 tropism. CONCLUSION: We have validated a new recombinant virus phenotypic assay for determining HIV-1 tropism using both plasma and cell samples from patients who are candidates for treatment with CCR5 antagonists. PMID- 20015685 TI - Impaired sleep quality in fibromyalgia: Detection and quantification with ECG based cardiopulmonary coupling spectrograms. PMID- 20015686 TI - Bioinformatics and molecular modelling approaches to GPCR oligomerization. AB - The elusive nature of the structure and function of the G-protein coupled receptor (GPCR) dimer or oligomer has led to a variety of computational studies, most of which have been directed primarily towards understanding structure. Here we review some of the recent studies based on sequence analysis and docking experiments and the recent developments in GPCR structure that have underpinned dimerization studies. In addition, we review recent nanosecond molecular dynamics simulations and coarse-grained methods for investigating the dynamic consequences of dimerization. The strengths and weaknesses of these complementary methods are discussed. The consensus of a variety of studies is that several transmembrane helices are involved in the dimerization/oligomerization interface(s); computation has been particularly effective in elucidating the experiments that seem to indicate a key role for transmembrane helix 4. PMID- 20015688 TI - Opening the crypt: current facts and hypotheses on the function of cryptopatches. AB - Cryptopatches, small aggregates of lymphoid cells found in the intestinal lamina propria, have been assigned many functions specific to gut immunity. Populated with seemingly immature lymphoid cells and dendritic cells, it has been suggested that cryptopatches maturate intraepithelial lymphocytes, Th17 cells, IL-22 producing NKp46(+) cells, and lymphoid tissues in response to the gut microbiota. Some of these issues, however, remain hotly debated. Therefore, cryptopatches are coming to the forefront of gut immunology and warrant a comprehensive discussion of their role in the development of the immune system. PMID- 20015687 TI - Oligomerization of G-protein-coupled receptors: a reality. PMID- 20015689 TI - Anatomy for nurses: providing students with the best learning experience. AB - A good understanding of human anatomy is clinically relevant for many health care professionals. A detailed knowledge of where, as well as how, to palpate, percuss or auscultate is essential for adequate assessment, provision of treatment and ongoing care of patients. Thus, the linking of human anatomy and clinical assessment is essential. However, Australian nursing students are often excluded from experiences, such as exposure to cadaveric material, which might enable them to gain a good working knowledge of internal human anatomy. Herein systematic student feedback from nursing students who were provided with a single exposure to prosected human anatomical material and were able to manipulate and interact with that material is presented, in the context of the integrative educational aims and goals of this learning and teaching activity. Thus, this study aims to add to the literature informing ongoing dialog regarding the best set of tools to teach anatomy to students of nursing. PMID- 20015690 TI - Learning features in computer simulation skills training. AB - New simulation tools imply new opportunities to teach skills and train health care professionals. The aim of this study was to investigate the learning gained from computer simulation skills training. The study was designed for optimal educational settings, which benefit student-centred learning. Twenty-four second year undergraduate nursing students practised intravenous catheterization with the computer simulation program CathSim. Questionnaires were answered before and after the skills training, and after the skills examination. When using CathSim, the students appreciated the variation in patient cases, the immediate feedback, and a better understanding of anatomy, but they missed having an arm model to hold. We concluded that CathSim was useful in the students' learning process and skills training when appropriately integrated into the curriculum. Learning features to be aware of when organizing curricula with simulators are motivation, realism, variation, meaningfulness and feedback. PMID- 20015691 TI - Nurse education in a resource limited environment: an evaluation of an educational teaching package on intramuscular injections, in Blantyre, Malawi. AB - This paper describes the implementation and evaluation of a pilot nursing education package of best practice guidelines involving intramuscular injections in a developing world setting. We briefly examine what is known about intramuscular injection techniques through reviewing the evidence on the topic, and disseminate best practice guidelines, in order to update and improve nursing technique in the paediatric departments of The Queen Elizabeth Central Hospital. METHODOLOGY: A teaching package of safe intramuscular injection techniques was implemented in October/November 2008. All paediatric wards were involved, and consisted of a 30 min structured package, with theory and then practical sessions. The effectiveness of the intervention was evaluated by a knowledge based questionnaires and non-participant observations of intramuscular injections taking place, to see if new techniques were being used. RESULTS: Theory based learning tested was at a high level and showed the nurses had a good scientific and evidence based understanding for their practice change. A convenience sample of 223 intramuscular injections was observed over December and January. Overall 188 (84.3%) took place in the thigh, the remainder in other locations. CONCLUSIONS: Theory with practical teaching can have an impact upon the care given, possibly reducing the complications associated with intramuscular injections. Further plans include ensuring sustainability of the programme and extension of the project to other developing countries. PMID- 20015692 TI - Patients perspective on endocarditis--an intermezzo in life. AB - BACKGROUND: Infective endocaditis (IE) is a life threatening disease with a mortality rate of 20-25%. There have been no previous reports, which describe the experience of health of patients suffering from IE. AIM: This study aimed to describe patients with IE's experience of health. This was done by having patients describe their experiences both before and during hospital admission, their experience of their physical symptoms, and their expectations for future health. METHODS: Qualitative interviews were conducted with 10 patients with IE (age 27-75) and transcribed. The analysis consisted of three levels. RESULTS: The overall concept that emerged was that IE is perceived as an intermezzo in life. The interviewees explain that a sudden unexpected physical change occurs that is difficult to understand and interpret. The concept "intermezzo in life" can be expounded as the following themes: Presage and appearance of IE, Reaction to IE, Living through IE, The little life with IE, Body change and Loved ones at a distance. CONCLUSION: IE is found to be a time of uncertainty and impenetrableness, where many things change, not least the perception of the body and self. PMID- 20015693 TI - Clinical activity of pemphigus vulgaris relates to IgE autoantibodies against desmoglein 3. AB - Pemphigus vulgaris (PV) is a severe autoimmune bullous skin disease and is primarily associated with IgG against desmoglein 3 (dsg3), a desmosomal adhesion protein. In light of the recent association of autoreactive T helper (Th) 2 cells with active PV, the present study sought to relate the occurrence of Th2 regulated dsg3-specific autoantibody subtypes, i.e. IgE and IgG4, in 93 well characterized PV patients. Patients with acute onset PV (n=37) showed the highest concentrations of serum IgE and IgG4 autoantibodies, which were significantly lower in PV patients in remission (n=14). Furthermore, there was a strong correlation between dsg3-reactive IgE and IgG4 in acute onset, but not in chronic active (n=42) or remittent patients. Additionally, intercellular IgE deposits were detected in the epidermis of acute onset PV. Thus, dsg3-specific IgE and IgG4 autoantibodies are related to acute onset disease which provides additional support to the concept that PV is a Th2-driven autoimmune disorder. PMID- 20015694 TI - Suppressive influences of IFN-alpha on IL-17 expression in human CD4+ T cells. AB - We examined the direct effects of IFN-alpha on the development of Th17 with a system using immobilized anti-CD3, which permits activation of CD4+ T cells in the complete absence of accessory cells. Highly purified CD4+ T cells obtained from healthy donors were stimulated with immobilized anti-CD3 with or without IFN alpha. IFN-alpha suppressed the production of IL-17 of immobilized anti-CD3 stimulated CD4+ T cells in a dose-response manner. Accordingly, IFN-alpha inhibited IL-17 mRNA expression in immobilized anti-CD3-stimulated CD4+ T cells. IFN-alpha did not affect the production of TGF-beta or IL-6, but inhibited RORC mRNA expression of anti-CD3-stimulated CD4+ T cells. These results indicate that IFN-alpha suppresses IL-17 expression and Th17 differentiation through down regulation of RORC mRNA expression. It is therefore suggested that these effects might play a role in the mode of action of IFN-alpha in the treatment of various inflammatory diseases. PMID- 20015695 TI - Sunitinib impairs the proliferation and function of human peripheral T cell and prevents T-cell-mediated immune response in mice. AB - Sunitinib (sunitinib malate; SU11248; SUTENT) is a novel multi-targeted receptor tyrosine kinase inhibitor currently approved for the treatment of metastatic renal cell carcinoma. To analyze the possible use of this compound in combination with immunotherapeutic approaches, we investigated the effects of sunitinib on the human peripheral T cells and the induction of primary immune responses in mice. Sunitinib inhibited the proliferation of primary human T cells from normal healthy volunteers as well as from renal cell carcinoma (RCC) and other cancer patients. The inhibition was recoverable after drug withdrawal because sunitinib did not induce T-cell apoptosis even at 0.8 muM. In addition, sunitinib led to accumulation in G(0)/G(1) phase of the cell cycle, inhibition of cytokine production, downregulation of activation markers expression and blockade of Zap 70 signaling in the T cells. Sunitinib significantly reduced the ear swelling induced by picryl chloride in mice. In light of these findings, the effects of sunitinib on the immune system should be emphasized for the therapy of metastatic renal cell carcinoma patients to avoid the impairment of T lymphocytes. PMID- 20015697 TI - Increased trapezius pain sensitivity is not associated with increased tissue hardness. AB - Fatiguing exercise can affect muscle pain sensitivity and muscle hardness, as seen with work-related neck and shoulder pain. Objective methods to assess muscle pain sensitivity are important because the reliability of manual assessment is generally poor. The aim of this study was (1) to compare coexistence of tender points identified by manual palpation and pressure algometry or hardness assessments and (2) to examine the influence of exercise on muscle pain sensitivity and hardness. Fourteen sites in the upper trapezius muscle were selected for assessments in 12 healthy subjects. Pressure pain thresholds and muscle hardness were examined by computer-controlled pressure algometry at baseline, immediately after static or dynamic exercise, and 20 minutes after static or dynamic exercise. Before recording of pressure pain thresholds, the trapezius muscle was examined for tender points by manual palpation. Two sites with low pressure pain thresholds were typical locations for tender points, and these were the least hard sites. However, manually detected tender points were often (29%) not colocalized with most sensitive sites according to the pressure algometry. A heterogeneous distribution of pressure pain sensitivity and muscle hardness was found in the upper trapezius. The short duration of exercise until exhaustion did not change muscle sensitivity or muscle hardness in asymptomatic muscles. PERSPECTIVE: This study confirms clinical findings with heterogeniosity in pain sensitivity and hardness across the upper trapezius muscle. Developments of new techniques that objectively can identify tender points are important, but thus far, manual palpation is best clinical practice. PMID- 20015696 TI - Smoking status and pain level among head and neck cancer patients. AB - Smoking is a risk factor for cancer of the upper aerodigestive tract with recidivism rates high even after diagnosis. Nicotine, a major product in tobacco, is a complex drug with multiple characteristics including analgesic properties. The goal of the study was to examine pain levels in the context of smoking status among patients recently diagnosed with cancer of the upper aerodigestive tract who have not yet received any treatment including radiation, surgery, or chemotherapy. A convenience sample of 112 newly diagnosed head and neck cancer patients (78 men and 34 women) was recruited from clinics at the University of Florida. Smoking rates were: 32% never smoked, 34% former smokers, 34% current smokers. Among current smokers, 62% reported plans to quit in the next 3 months and 38% had tried to quit more than 3 times in the past 5 years. Current smokers reported higher general (sensory and affective) and oral pain levels (spontaneous and functional) and pain-related interference than did never and former smokers (all F's > 8. and P's < .0001) even after controlling for stage of diagnosis. In addition, current smokers reported significantly greater interference from the pain (F(2,73) = 10.5 P < .0001). PERSPECTIVE: This study highlights the importance of understanding self-reported pain in cancer patients who continue to smoke. When pain is elevated, smokers may be motivated to use tobacco as a means of reducing pain, which in turn reinforces smoking behavior. Tobacco cessation programs should include pain management as a component of treatment. PMID- 20015698 TI - A principal components analysis of negative affect-related constructs relevant to pain: evidence for a three component structure. AB - A number of negative affect-related constructs are important in pain. Some are general, such as anxiety, depression and negative affectivity, whereas others are more specifically pain-related (eg, fear of pain, pain anxiety, and pain catastrophizing). In addition, some more specific fear-related constructs, such as anxiety sensitivity, illness/injury sensitivity, and fear of negative evaluation have emerged as important to pain. Although these various constructs are considered conceptually separate, there is likely to be overlap between them. Since the extent of this overlap is unknown, the aim of the current study was to investigate these constructs in 1 sample in order to identify their common and unique features. Frequently used psychological measures were completed by 508 pain-free participants. Principal components analysis resulted in the extraction of three components: 1) General distress; 2) Fear of pain from injury/insult; and 3) Cognitive intrusion of pain. The results presented here suggest that there is indeed commonality between constructs, which may be due to either an overlap between items within measures or to close conceptual relatedness. The implications of these core dimensions are discussed with reference to future research and theory. PERSPECTIVE: This article explores the relationships between various negative-affect pain-related measures and discusses the results from a principal components analysis. The findings show that some questionnaires may measure the same latent construct. A measure could be developed to measure these 3 core components more concisely for both clinical and research purposes. PMID- 20015699 TI - Spinal nerve ligation in mouse upregulates TRPV1 heat function in injured IB4 positive nociceptors. AB - Peripheral nerve injury leads to neuropathic pain, but the underlying mechanisms are not clear. The TRPV1 channel expressed by nociceptors is one receptor for noxious heat and inflammatory molecules. Lumbar 4 (L4) spinal nerve ligation (SNL) in mice induced persistent heat hyperalgesia 4 to 10 days after injury. The heat hypersensitivity was completely reversed by the TRPV1 antagonist A-425619. Furthermore, DRG neurons were isolated from the injured L4 ganglia or adjacent L3 ganglia 4 to 10 days after L4 SNL. Whole-cell patch-clamp recordings were performed and heat stimuli (22 degrees C to 50 degrees C/3 s) were applied to the soma. Neurons were classified by soma size and isolectin-B4 (IB4) binding. Among directly injured L4 neurons, SNL increased the percentage of small-diameter IB4 positive neurons that were heat-sensitive from 13% (naive controls) to 56% and conversely decreased the proportion of small IB4-negative neurons that were heat sensitive from 66% (naive controls) to 34%. There was no change in IB4 binding in neurons from the injured ganglia. Surprisingly, in neurons from the adjacent L3 ganglia, SNL had no effect on the heat responsiveness of either IB4-positive or negative small neurons. Also, SNL had no effect on heat responses in medium-large diameter neurons from either the injured or adjacent ganglia. PERSPECTIVE: TRPV1 function is upregulated in IB4-positive sensory neurons, and TRPV1 is responsible for the behavioral heat hypersensitivity in the spinal nerve ligation model. Because IB4-positive neurons may contribute to the emotional perception of pain, TRPV1 antagonists, targeting both sensory and affective pain components, could have broad analgesic effects. PMID- 20015701 TI - Tonic pain abolishes cortical habituation of visual evoked potentials in healthy subjects. AB - We investigated changes in visual cortex excitability by analyzing visual evoked potential (VEP) habituation in healthy subjects during tonic pain evoked by the cold-pressor test (CPT). We tested VEP amplitude habituation (slope of the linear regression line for N1-P1 amplitude from the 1st to 6th block of 100 sweeps) in 19 healthy volunteers during 4 experimental conditions: baseline; no-pain (hand held in warm water, 25 degrees C); pain (hand held in cold water, 2-4 degrees C); and the after-effects of tonic pain. During baseline and no-pain sessions, VEPs habituated normally across the 6 consecutive blocks (mean slope -.28 and -.18%), whereas during pain and its after-effects they failed to decrease (0%, and .11%). Tonic pain induced by the CPT abolishes normal VEP habituation and the lack of habituation persists after the CPT is stopped. Tonic pain probably abolishes VEP habituation by acting on brainstem neural structures which modulate thalamo-cortical activation thereby changing visual cortex excitability. PERSPECTIVE: This study shows that tonic pain alters visual cortex excitability, a brain region unrelated to pain processing. These changes probably reflect defensive strategies against pain. Extending the study from healthy volunteers to patients with migraine between attacks would offer the opportunity to investigate visual cortical excitability under conditions when baseline habituation is absent. PMID- 20015700 TI - ASIC1 and ASIC3 play different roles in the development of Hyperalgesia after inflammatory muscle injury. AB - Acid-sensing ion channels (ASICs) respond to acidosis that normally occurs after inflammation. We examined the expression of ASIC1, ASIC2, and ASIC3 mRNAs in lumbar dorsal root ganglion neurons before and 24 hours after carrageenan-induced muscle inflammation. Muscle inflammation causes bilateral increases of ASIC2 and ASIC3 but not ASIC1 (neither ASIC1a nor ASIC1b) mRNA, suggesting differential regulation of ASIC1 versus ASIC2 and ASIC3 mRNA. Similar mRNA increases were observed after inflammation in knockout mice: ASIC2 mRNA increases in ASIC3-/- mice; ASIC2 and ASIC3 mRNAs increase in ASIC1-/- mice. Prior behavioral studies in ASIC3-/- mice showed deficits in secondary hyperalgesia (increased response to noxious stimuli outside the site of injury) but not primary hyperalgesia (increased response to noxious stimuli at the site of injury). In this study, we show that ASIC1-/- mice do not develop primary muscle hyperalgesia but develop secondary paw hyperalgesia. In contrast, and as expected, ASIC3-/- mice develop primary muscle hyperalgesia but do not develop secondary paw hyperalgesia. The pharmacological utility of the nonselective ASIC inhibitor A-317567, given locally, was tested. A-317567 reverses both the primary and the secondary hyperalgesia induced by carrageenan muscle inflammation. Thus, peripherally located ASIC1 and ASIC3 play different roles in the development of hyperalgesia after muscle inflammation. PERSPECTIVE: This study shows changes in ASIC mRNA expression and behavioral hyperalgesia of C57Bl/6 (wild type), ASIC1-/-, and ASIC3-/- mice before and after the induction of muscle inflammation. A-317567 was effective in reversing hyperalgesia in these animals, suggesting the potential of ASICs as therapeutic targets for muscle inflammatory pain. PMID- 20015703 TI - Emotional regulation and acute pain perception in women. AB - Emotional regulation is an important variable in the experience of pain. Currently, there are no experimental investigations of the relation between emotional regulation and pain. The goal of the present study work was to analyze differences in pain perception and mood generated by the cold-pressor (CPT) experimental paradigm in women with high and low emotional regulation. Two groups of women were formed as a function of their level of emotional regulation: women with high emotional repair (N = 24) and women with low emotional repair (N = 28), all of whom performed the CPT. The results show that the women with a high score in emotional repair reported having experienced less sensory pain and affective pain during the immersion, as well as a more positive affective state before beginning the task. During the experimental task, they also reported a better mood, thus displaying lower impact of the experience of pain. PERSPECTIVE: Emotional regulation is proposed as a key element to manage the emotional reaction that accompanies the experience of acute pain experimentally induced by the CPT experimental paradigm in a sample of healthy women. PMID- 20015702 TI - Evaluation of nurses' self-insight into their pain assessment and treatment decisions. AB - Research generally indicates that providers demonstrate modest insight into their clinical decision processes. In a previous study utilizing virtual human (VH) technology, we found that patient demographic characteristics and facial expressions of pain were statistically significant predictors of many nurses' pain-related decisions. The current study examined the correspondence between the statistically identified and self-reported influences of contextual information on pain-related decisions. Fifty-four nurses viewed vignettes containing a video of a VH patient and text describing a postsurgical context. VH sex, race, age, and facial expression varied across vignettes. Participants made pain-assessment and treatment decisions on visual analogue scales. Participants subsequently indicated the information they relied on when making decisions. None of the participants reported using VH sex, race, or age in their decision process. Statistical modeling indicated that 28 to 54% of participants (depending on the decision) used VH demographic cues. 76% of participants demonstrated concordance between their reported and actual use of the VH facial expression cue. Vital signs, text-based clinical summary, and VH movement were also reported as influential factors. These data suggest that biases may be prominent in practitioner decision-making about pain, but that providers have minimal awareness of and/or a lack of willingness to acknowledge this bias. PERSPECTIVE: The current study highlights the complexity of provider decision-making about pain management. The VH technology could be used in future research and education applications aimed at improving the care of all persons in pain. PMID- 20015704 TI - Capsaicin-induced central sensitization evokes segmental increases in trigger point sensitivity in humans. AB - This study investigated whether inducing central sensitization evokes segmental increases in trigger point pressure sensitivity. We evoked central sensitization at the C(5) segment and validated its presence via mechanical cutaneous sensitivity (brush allodynia) testing. Trigger point pressure sensitivity was quantified using the pain pressure threshold (PPT) value. A 50 cm(2) area of the C(5) dermatome at the right lateral elbow was pretreated with 45 degrees heat for 10 minutes. Test subjects (n = 20) then received topical capsaicin cream (0.075%; Medicis, Toronto, Canada) to the C(5) dermatome, whereas control subjects (n = 20) received a topical placebo cream (Biotherm Massage, Montreal, Canada). PPT readings were recorded from the infraspinatus (C(5,6)) and gluteus medius (L(4,5)S(1)) trigger points at zero (pre-intervention), 10, 20, and 30 minutes after intervention; all PPT readings were normalized to pre-intervention (baseline) values. The difference between the PPT readings at the 2 trigger point sites represents the direct influence of segmental mechanisms on the trigger point sensitivity at the infraspinatus site (PPT(seg)). Test subjects demonstrated statistically significant increases in Total Allodynia scores and significant decreases in PPT(seg) at 10, 20, and 30 minutes after application, when compared with control subjects. These results demonstrate that increases in central sensitization evoke increases in trigger point pressure sensitivity in segmentally related muscles. PERSPECTIVE: Myofascial pain is the most common form of musculoskeletal pain. Myofascial trigger points play an important role in the clinical manifestation of myofascial pain syndrome. Elucidating the role of central sensitization in the pathophysiology of trigger points is fundamental to developing optimal strategies in the management of myofascial pain syndrome. PMID- 20015705 TI - Preliminary psychometric properties of the Chinese version of the Chronic Pain Coping Inventory (ChCPCI) in a Hong Kong Chinese population. AB - The Chronic Pain Coping Inventory (CPCI) is a frequently used measure that assesses 8 categories of coping strategies that patients might use to cope with chronic pain. Despite its good psychometric properties and widespread use, the instrument has not been tested for its applicability and validity in non-Western populations, such as among Chinese. This study evaluated the reliability and validity of a Chinese translation of the 42-item CPCI (ChCPCI-42) in a sample of Chinese patients with chronic pain (n = 208). In addition to the ChCPCI-42, the patients were administered the Chronic Pain Grade (CPG) questionnaire, the Pain Catastrophizing Scale (PCS), the Centre for Epidemiological Studies-Depression Scale (CES-D), and questions assessing sociodemographic characteristics. Results of confirmatory factor analyses revealed that of the ChCPCI-42 8 scales, 6 demonstrated acceptable-to-good data-model fit (CFI >or= 0.90) and 2 demonstrated medium fit (CFI >or= 0.85). The 8 scales demonstrated adequate to good internal consistency (Cronbach alpha, 0.69 to 0.79) and correlated with CES-D, PCS, pain intensity, and disability in expected directions. Results of hierarchical multiple regression analyses showed that the ChCPCI-42 scales predicted concurrent depression (F (8,177) = 3.07, P < .01) and pain disability (F (1, 179) = 4.35, P < .001) scores, the Task Persistence scale being the strongest unique predictor among the 8 scales. The findings support the factorial validity and reliability of a 42-item CPCI that can be used among Chinese patients with chronic pain. PERSPECTIVE: The report outlines the first validation of the CPCI for use in Hong Kong Chinese. This makes available a suitable instrument for chronic pain research in the Southern Chinese population and will help to elucidate similarities and differences in pain coping between Chinese and other ethnic groups. PMID- 20015706 TI - The impact of adolescent chronic pain on functioning: disentangling the complex role of anxiety. AB - A number of adolescents with chronic pain have clinically significant disability across physical, social, and academic activities, and pain severity only explains a portion of the variance in functioning. Thus, it is important to identify therapeutic options to improve adolescents' functioning. In contrast to studies with adults with chronic pain, research in pediatric pain has not consistently found anxiety to be a good predictor of pain-related disability. The present study evaluated pain, anxiety, and functioning in 222 adolescents with chronic pain. Results indicated that pain was consistently and linearly related to disability across measures of physical and social functioning, school attendance, and physician visits. The relation between anxiety and functioning was complex; increased anxiety was related to poorer physical and social functioning and was related to fewer physician visits, although it was not associated with school attendance. Additional analyses revealed that anxiety serves to moderate the relation between pain and functioning. Specifically, at high anxiety, pain was not related to functioning, but at low anxiety, pain consistently predicted disability. In other words, highly anxious adolescents were functioning poorly regardless of the level of pain. The moderating role of anxiety highlights a number of research and clinical possibilities to explore with adolescents with chronic pain-related disability. PERSPECTIVE: Data suggest that high anxiety is associated with poor functioning irrespective of pain intensity. At low anxiety, higher pain predicted greater disability. Anxiety is important to assess when investigating potential reasons for pain-related disability. PMID- 20015707 TI - Thermal nociception is decreased by hypocretin-1 and an adenosine A1 receptor agonist microinjected into the pontine reticular formation of Sprague Dawley rat. AB - Clinical and preclinical data concur that sleep disruption causes hyperalgesia, but the brain mechanisms through which sleep and pain interact remain poorly understood. Evidence that pontine components of the ascending reticular activating system modulate sleep and nociception encouraged the present study testing the hypothesis that hypocretin-1 (orexin-A) and an adenosine receptor agonist administered into the pontine reticular nucleus, oral part (PnO) each alter thermal nociception. Adult male rats (n = 23) were implanted with microinjection guide tubes aimed for the PnO. The PnO was microinjected with saline (control), hypocretin-1, the adenosine A(1) receptor agonist N(6)-p sulfophenyladenosine (SPA), the hypocretin receptor-1 antagonist N-(2-Methyl-6 benzoxazolyl)-N''-1,5-naphthyridin-4-yl-urea (SB-334867), and hypocretin-1 plus SB-334867. As an index of antinociceptive behavior, the latency (in seconds) to paw withdrawal away from a thermal stimulus was measured following each microinjection. Compared to control, antinociception was significantly increased by hypocretin-1 and by SPA. SB-334867 increased nociceptive responsiveness, and administration of hypocretin-1 plus SB-334867 blocked the antinociception caused by hypocretin-1. These results suggest for the first time that hypocretin receptors in rat PnO modulate nociception. PERSPECTIVE: Widely distributed and overlapping neural networks regulate states of sleep and pain. Specifying the brain regions and neurotransmitters through which pain and sleep interact is an essential step for developing adjunctive therapies that diminish pain without disrupting states of sleep and wakefulness. PMID- 20015708 TI - Translation to Portuguese and validation of the Douleur Neuropathique 4 questionnaire. AB - The Douleur Neuropathique 4 (DN4) questionnaire was developed by the French Neuropathic Pain Group and is a simple and objective tool, with the ability to distinguish nociceptive from neuropathic pain. The purpose of this work was to validate the DN4 questionnaire in the Portuguese language in order to allow its use in clinical and research settings. A double-blind, accuracy study was conducted, consisting of translation, back-translation, literal evaluation, semantic equivalence, and communication with the target population. The Portuguese version of the questionnaire was applied in a sample of 101 patients with neuropathic (N = 42) or nociceptive pain (N = 59), ranked according to medical diagnosis. The reproducibility, reliability and validity of the instrument were analyzed, and showed a high diagnostic power for this version of the DN4 questionnaire. The Portuguese version of the DN4 questionnaire presented good validity and reliability, allowing it to identify neuropathic pain and neuropathic characteristics of mixed pain syndromes. PERSPECTIVE: This article presents the first validated neuropathic pain questionnaire in the Portuguese language and represents a useful tool in the assessment of neuropathic pain both in the clinical setting and in population-based studies. The sensible and quick format of this instrument are key factors that will contribute to its widespread use, permitting a true recognition of patients with neuropathic pain. PMID- 20015709 TI - Application of displacement chromatography for the analysis of a lipid raft proteome. AB - Defining membrane proteomes is fundamental to understand the role of membrane proteins in biological processes and to find new targets for drug development. Usually multidimensional chromatography using step or gradient elution is applied for the separation of tryptic peptides of membrane proteins prior to their mass spectrometric analysis. Displacement chromatography (DC) offers several advantages that are helpful for proteome analysis. However, DC has so far been applied for proteomic investigations only in few cases. In this study we therefore applied DC in a multidimensional LC-MS approach for the separation and identification of membrane proteins located in cholesterol-enriched membrane microdomains (lipid rafts) obtained from rat kidney by density gradient centrifugation. The tryptic peptides were separated on a cation-exchange column in the displacement mode with spermine used as displacer. Fractions obtained from DC were analyzed using an HPLC-chip system coupled to an electrospray-ionization ion-trap mass spectrometer. This procedure yielded more than 400 highly significant peptide spectrum matches and led to the identification of more than 140 reliable protein hits within an established rat kidney lipid raft proteome. The majority of identified proteins were membrane proteins. In sum, our results demonstrate that DC is a suitable alternative to gradient elution separations for the identification of proteins via a multidimensional LC-MS approach. PMID- 20015710 TI - Simultaneous quantification of haemoglobin adducts of ethylene oxide, propylene oxide, acrylonitrile, acrylamide and glycidamide in human blood by isotope dilution GC/NCI-MS/MS. AB - Haemoglobin adducts are highly valuable biomarkers of cumulative exposure to carcinogenic substances. We have developed and applied an analytical method for the simultaneous quantification of five haemoglobin adducts of important occupational and environmental carcinogens. The N-terminal adducts were determined with gas chromatography as pentafluorophenylthiohydantoine derivatives according to the modified Edman-procedure and subsequent acetonization of the glycidamide adduct N-(R,S)-2-hydroxy-2-carbamoylethylvaline (GAVal). The use of self-synthesized labelled internal standards in combination with tandem mass spectrometry using negative chemical ionisation guarantees both high accuracy and sensitivity of our determination. The limit of detection for N-2 hydroxyethylvaline (HEVal), N-(R,S)-2-hydroxypropylvaline (HPVal), N-2 carbamoylethylvaline (AAVal) and N-(R,S)-2-hydroxy-2-carbamoylethylvaline (GAVal) was 2 pmol/g globin, for N-2-cyanoethylvaline (CEVal) it was determined as 0.5 pmol/g globin, which was sufficient to determine the background levels of these adducts in the non-smoking general population. The between-day-precision for all analytes using a human blood sample as quality control material ranged from 4.7 to 12.3%. We investigated blood samples of a small group (n=104) of non-smoking persons of the general population for the background levels of these haemoglobin adducts. The median values for HEVal, HPVal, CEVal, AAVal and GAVal in a group of 92 non-smoking persons were 18.1, 4.1, <0.5, 29.9 and 35.2 pmol/g globin, respectively. The adduct levels in 12 persons reporting exposure to passive smoke at home were similar for most adducts with median values of 17.2, 4.1, 1.0, 24.9 and 29.7 pmol/g globin for HEVal, HPVal, CEVal, AAVal and GAVal, respectively. Our results point to an elevated uptake of acrylonitrile caused by passive smoking as indicated by higher levels of the corresponding haemoglobin adduct CEVal. PMID- 20015711 TI - Determination of D-saccharic acid-1,4-lactone from brewed kombucha broth by high performance capillary electrophoresis. AB - Kombucha is a health tonic. D-saccharic acid-1,4-lactone (DSL), a component of kombucha, inhibits the activity of glucuronidase, an enzyme indirectly related with cancers. To date, there is no efficient method to determine the content of DSL in kombucha samples. In this paper, we report a rapid and simple method for the separation and determination of DSL in kombucha samples, using the high performance capillary electrophoresis (HPCE) method with diode array detection (DAD). With optimized conditions, DSL can be separated in a 50 cm length capillary at a separation voltage of 20 kV in 40 mmol/L borax buffer (pH 6.5) containing 30 mmol/L SDS and 15% methanol (v/v). Quantitative evaluation of DSL was determined by ultraviolet absorption at lambda=190 nm. The relationship between the peak areas and the DSL concentrations, in a specified working range with linear response, was determined by first-order polynomial regression over the range 50-1500 microg/mL with a detection limit of 17.5 microg/mL. Our method demonstrated excellent reproducibility and accuracy with relative standard deviations (RSD) of less than 5% DSL content (n=5). This is the first report to determine DSL by HPCE. We have successfully applied this method to determine DSL in kombucha samples in various fermented conditions. PMID- 20015712 TI - The 'vacsplint' for hands. PMID- 20015713 TI - Ludwig's angina as an extremely unusual complication for direct microlaryngoscopy. AB - BACKGROUND: An extremely rare case that to our knowledge has not been reported before is described, in which a patient had a Ludwig's angina as a complication of direct microlaryngoscopy. METHODS: We report a Ludwig's angina after a direct microlaryngoscopy for a Reinke's edema, due to erosion on the internal face of the mandible produced by compression of the laryngoscope. RESULTS: The patient underwent placement of 2 drainages, intraoral and cervical, and several incisions on the floor of the mouth, with intravenous corticosteroids and antibiotics and with resolution of the illness without performing tracheostomy. CONCLUSIONS: Ludwig's angina is an extremely rare complication of microlaryngoscopy, but it is potentially life-threatening. Early diagnosis and treatment resulted in survival of the patient without complications. PMID- 20015714 TI - Novel presentation of a fourth branchial cleft anomaly in a male infant. AB - Fourth branchial cleft anomalies are rare congenital disorders of the neck. We describe a case involving a unique presentation of this entity as well as a review of the literature concerning its management. PMID- 20015715 TI - An unusual case of ingestion of a moth cocoon in a 14-month-old girl. AB - We present a case report of a 14-month-old girl who ingested a moth cocoon, which resulted in dramatic symptoms of irritability, drooling, and anorexia. Direct laryngoscopy, bronchoscopy, and esophagoscopy under general anesthesia revealed copious, tenaciously adherent, barbed hairs embedded in her tongue and buccal mucosa. Removal of the hairs with irrigation, suction, and brushing was unsuccessful and was eventually abandoned. In the following 48 hours, the girl recovered uneventfully with supportive care. The hairs were subsequently identified as coming from the hickory tussock moth (Lepidoptera: Arctiidae: Lophocampa caryae), which is ubiquitously distributed throughout much of North America. This is the first detailed case report of ingestion of an L caryae cocoon. PMID- 20015716 TI - Intracranial spread of chronic middle ear suppuration. AB - OBJECTIVE: The aim of the study was to review the pathogenesis and the result of management of the intracranial complications of chronic middle ear suppuration. METHODS: This was a retrospective review of charts of 32 cases with intracranial complications due to chronic middle ear infection managed between 1993 and 2007. The symptoms, clinical findings, and medical and surgical management were reviewed and analyzed. RESULTS: There were 10 (31.2%) patients in the age group of 0 to 10 years, 9 (28.1%) patients in the age group of 11 to 18 years, and 13 (40.6%) patients older than 18 years. Males were involved twice as much as females. Among the 32 patients, 18 (56.3%) had a single intracranial complication, whereas 14 (43.7%) had multiple intracranial complications. Among all the intracranial complications in the 32 patients, otitic meningitis was the commonest intracranial complication and was seen in 14 (43.7%) patients; it was followed by lateral sinus thrombosis in 10 (31.2%), cerebellar abscess in 6 (18.7%), epidural abscess in 7 (21.8%), and perisinus abscess in 5 (15.6%). Other less common but serious intracranial complications encountered were cerebral abscess and interhemispheric abscess in 2 (6.2%) each, and subdural abscess, otitic hydrocephalus, and otogenic cavernous sinus thrombosis in 1 (3.1%) each. Upon admission, all patients received a combination of parenteral antibiotics. Canal wall down mastoidectomy was performed in all but 1 patient. In addition, lateral sinus was explored in 13 (40.6%) and cerebellar abscesses were drained in 5 (15.6%) patients. The overall mortality rate of 31.2% was found in our series. CONCLUSION: The prognosis was worse with delayed presentation because of overwhelming intracranial infection due to multiple pathways of extension from chronic otitis media. Infected thrombus in the dural venous sinus should be removed to prevent dissemination of septic emboli. PMID- 20015717 TI - Intratumoral delivery of docetaxel enhances antitumor activity of Ad-p53 in murine head and neck cancer xenograft model. AB - PURPOSE: The aim of this study is to determine the ability of intratumorally delivered docetaxel to enhance the antitumor activity of adenovirus-mediated delivery of p53 (Ad-p53) in murine head and neck cancer xenograft model. MATERIALS AND METHODS: A xenograft head and neck squamous cell carcinoma mouse model was used. Mice were randomized into 4 groups of 6 mice receiving 6 weeks of biweekly intratumoral injection of (a) diluent, (b) Ad-p53 (1 x 10(10) viral particles per injection), (c) docetaxel (1 mg/kg per injection), and (d) combination of Ad-p53 (1 x 10(10) viral particles per injection) and docetaxel (1 mg/kg per injection). Tumor size, weight, toxicity, and overall and disease-free survival rates were determined. RESULTS: Intratumoral treatments with either docetaxel alone or Ad-p53 alone resulted in statistically significant antitumor activity and improved survival compared with control group. Furthermore, combined delivery of Ad-p53 and docetaxel resulted in a statistically significant reduction in tumor weight when compared to treatment with either Ad-p53 or docetaxel alone. CONCLUSION: Intratumoral delivery of docetaxel enhanced the antitumor effect of Ad-p53 in murine head and neck cancer xenograft model. The result of this preclinical in vivo study is promising and supports further clinical testing to evaluate efficacy of combined intratumoral docetaxel and Ad p53 in treatment of head and neck cancer. PMID- 20015718 TI - Necrotizing fasciitis secondary to bisphosphonate-induced osteonecrosis of the jaw. AB - Osteonecrosis of the jaw is an uncommon consequence of biphosphonate therapy. This has most commonly been a bone complication with little if any soft tissue involvement. An unusual case of necrotizing fasciitis with extensive soft tissue infection stemming from a prolonged case of osteonecrosis of the jaw presented. The management of this patient (aggressive surgical debridement and prolonged wound care) is reviewed as well as the review of the underlying processes. PMID- 20015719 TI - Localized amyloidosis of the nasal and paranasal mucosa: a rare pathology. PMID- 20015720 TI - Epiglottis reconstruction with free radial forearm flap after supraglottic laryngectomy. AB - A bilobed free radial forearm flap was designed to reconstruct a defect in the epiglottis and tongue base in 2 patients who underwent supraglottic laryngectomy. The flap was initially sutured in the shape of the epiglottis to prevent aspiration during deglutition. Six months after surgery, after a full course of radiation therapy, the flap had flattened and underwent atrophy, but the patients still had good voice production and were able to swallow well without any aspiration. Regardless of the final shape of the reconstructed epiglottis, it will suffice to prevent aspiration if the flap is large enough to occlude the tracheal outlet. PMID- 20015721 TI - Oculocardiac reflex during the endoscopic sinus surgery. AB - The oculocardiac reflex is developed by surgical or nonsurgical stimulations to the eyeball including manipulation of extraocular muscles. The authors came across a case of severe arrhythmia due to oculocardiac reflex caused by the stimulation of orbital tissue from the injury of lamina papyracea during endoscopic sinus surgery. Previous heart evaluations of the patient of this case suggested no abnormalities in the heart. This case shows that the oculocardiac reflex can occur during endoscopic sinus surgery but is not limited as the complication from ocular surgery. PMID- 20015722 TI - Endonasal laser-assisted microscopic dacryocystorhinostomy: surgical technique and follow-up results. AB - PURPOSE: Endonasal dacryocystorhinostomy is known as an increasingly attractive and effective approach for the surgical treatment of nasal duct obstruction with minimal complications and best cosmetic consequences. In a relatively large-scale case-series study over a 5-year period, we describe the surgical technique and 12 month follow-up results of microscopic laser dacryocystorhinostomy with particular regard to the effect of various pre-/postoperational factors (ie, patients' sex, age, symptoms chronicity, previous interventions, duration of silicone intubation) on the surgical outcome. MATERIALS AND METHODS: A total of 162 cases in 151 patients with chronic epiphora, mucocele, or recurrent episodes of dacryocystitis were included in the study. Endonasal laser dacryocystorhinostomy was performed using a surgical microscope with transcanalicular lacrimal sac illumination. The laser types used were potassium titanyl-phosphate and neodymium:yttrium-aluminum-garnet for ablation of nasal mucosa and application to bone, respectively. Patients were evaluated 6 months and 1 year later. Data were analyzed by chi(2) tests. RESULTS: There were no major complications during or after the operations. Complete cure occurred in 89.5% (after 6 months) and 74.2% (after 1 year) of the cases. Anatomical patency was shown by lacrimal system irrigation with fluorescein in 81.5% of the cases after the 12-month follow-up. It was found that patients younger than 55 years, with symptoms lasting less than 1 year, and without history of nasal problems, had significantly higher surgical success rates (P < .05). Moreover, rates of failure were significantly lower in cases whose canaliculi were intubated for 5 to 6 months (P < .05). CONCLUSIONS: Endonasal microscopic laser dacryocystorhinostomy is a safe and minimally invasive procedure with reasonable results. It has many advantages over external or other conventional approaches. Successful results could be further enhanced by more wisely selecting the patients and by silicone extubation after 6 months. PMID- 20015723 TI - Difference in outcome of botulinum toxin treatment of essential palatal tremor in children and adults. AB - PURPOSE: The objective of this clinical study was to investigate the history and clinical findings in 10 patients having an essential palatal tremor. Furthermore, a botulinum toxin A (BTA) therapy in 5 cases was carried out, and the outcome was analyzed. MATERIALS AND METHODS: Seven adult and 3 pediatric patients with essential palatal tremor were examined at presentation, before and after start of treatment, and every 3 months or when symptoms recurred. Findings were documented by endoscopic video recordings, electromyography, tympanometry, and ear canal microphone recording. The BTA injections were performed in local or general anesthesia, under elecromyographic guidance. RESULTS: The BTA therapy in all 5 patients was successful. Surprisingly, 2 of these patients, aged 10 and 6 years, remained in remission for several years after a single successful injection. CONCLUSION: Botulinum toxin therapy is a safe and effective treatment of essential palatal tremor and seems to be especially useful in pediatric patients. The long lasting effect in children hints toward a pathophysiologic difference between pediatric and adult essential palatal tremor. PMID- 20015724 TI - Tympanometry in infants with middle ear effusion having been identified using spiral computerized tomography. AB - PURPOSE: This study was carried out to evaluate the diagnostic value of 226 and 1000 Hz probe-tone tympanometry in infants with effusion in the middle ear. METHODS: For this study, we recruited 52 infants with ages 42 days to 6 months as clinical subjects in a hearing-ability screening program. After a spiral computerized tomography (CT) scan of the patients, we tested their hearing using tympanometry of 2 probe-tone frequencies: 226 and 1000 Hz. The patients were divided into 2 groups according to the results of the CT scan: group 1 patients had normal middle ears without fluid, and group 2 patients had ears with fluid. We recorded the tympanograms and their percentage of every type and compared the tympanometric results with CT to get the concordance rate between tympanometry and CT diagnose while obtaining the normal values of 1000 Hz tympanometric measures. RESULTS: The 226 Hz probe-tone tympanograms of middle ears with fluid differed greatly from those without. At 226 Hz, their tympanograms were single peaked tympanograms (51.06%), double-peaked tympanograms (44.68%), flat tympanograms (2.13%), and negative-pressure tympanograms (2.13%) for the group with normal middle ears, but single-peaked tympanograms (77.19%), double-peaked tympanograms (19.30%), and flat tympanograms (3.51%) for the group with middle ear effusion. The 1000 Hz probe-tone tympanograms included single-peaked or flat type tympanograms in both the normal middle ear group and the group with middle ear effusion. The group with normal middle ears was identified by spiral CT, and its tympanograms mostly had a single peak (97.87%) during 1000 Hz tympanometry. Tympanograms of the middle ear effusion group mostly had a flat curve (98.25%). When the Liden/Jerger classification system was used to evaluate the tympanograms, normal tympanograms were single-peaked, and flat tympanograms indicated middle ear effusion. According to this standard, the concordance rate between the 1000 Hz tympanometry (98.08%) and CT diagnosis was higher than when 226 Hz tympanometry (25%) (P < .05) was performed, and the value of kappa was equal to 0.961 between 1000 Hz tympanometry and CT diagnosis. CONCLUSIONS: In clinical practice, 1000 Hz tympanometry, not 226 Hz, is recommended to determine the presence of middle ear fluid in infants younger than 6 months. PMID- 20015726 TI - The dilemma of midline destructive lesions: a case series and diagnostic review. AB - BACKGROUND: Midline destructive lesions (MDLs) of the nose are a diagnostic dilemma due to an extensive differential diagnosis and vague presenting signs and symptoms. Etiologies may be neoplastic, autoimmune, traumatic, infectious, or unknown. STUDY DESIGN: Case series and review of the literature were done. METHODS: Medical records of 8 patients presenting with an MDL were reviewed. RESULTS: Each patient received nasal endoscopy, computed tomography scan of the sinuses, laboratory workup, culture (aerobes, anaerobes, fungus, and acid-fast bacilli), and biopsy with flow cytometry. Laboratory tests included complete blood count, basic metabolic panel, erythrocyte sedimentation rate, angiotensin converting enzyme, antineutrophil antibodies, rheumatoid factor, anti-Ro and anti La antibodies, Epstein-Barr virus antibodies, coccidiomycosis serology, HIV antibodies, fluorescent treponemal antibody absorption, classic antineutrophil cytoplasmic antibodies, perinuclear antineutrophil cytoplasmic antibody, proteinase 3, and myeloperoxidase. Choice of diagnostic study was individualized for each patient. Two patients were diagnosed with natural killer/T-cell lymphoma, 2 were diagnosed with Wegener's granulomatosis, and 4 remained idiopathic, despite the extensive workup. A diagnostic algorithm to aid in the approach to MDLs is presented. CONCLUSIONS: The diagnosis of MDLs remains difficult but is aided by a systematic approach and familiarity with multiple diagnostic techniques. It is imperative to take multiple tissue specimens from various sites, send them fresh, and communicate suspicion of lymphoma. Despite diagnostic advances and improved understanding of the diseases underlying MDLs, an etiology is often not identified. PMID- 20015725 TI - Schwannoma of the hard palate. AB - Schwannomas are rare, benign neoplasms that can arise from any cranial, peripheral, or autonomic nerve that contains Schwann cells. Approximately 25% to 45% of all schwannomas occur in the head and neck. They occur most commonly in the eighth cranial nerve, but it has been reported that 20% to 58% arise in the oral cavity, with approximately 10% of these located on the hard palate. We report a case of schwannoma of the hard palate, present important pathologic considerations for diagnosis, and provide a review of the literature regarding extracranial schwannomas. PMID- 20015727 TI - Lidocaine spray vs tetracaine solution for transnasal fiber-optic laryngoscopy. AB - STATEMENT OF PROBLEM: The aim of this study was to evaluate the efficacy of lidocaine spray 10%, compared with tetracaine 2% solution, as a local anesthetic for patients undergoing transnasal fiber-optic laryngoscopy. METHOD OF STUDY: A prospective study was conducted on patients undergoing transnasal fiber-optic laryngoscopy. Microsurgical sponges were applied in each side of the nose for 10 minutes before laryngoscopy. Patients were randomly classified into group A and group B, in which tetracaine 2% solution and lidocaine spray 10% were used, respectively. Patients were asked to evaluate the severity of pain during the procedure by a visual analog scale. Patients data, pain score, and potential complications were placed in a database and statistically assessed. MAIN RESULTS: Our series consisted of 48 patients. Statistical analysis showed significant lower mean nasal discomfort score in favor of the tetracaine group (2.29 vs 3.04 [P < .001]). No tetracaine complications or side effects occurred. PRINCIPAL CONCLUSION: Neurosurgical sponge application of tetracaine 2% solution is an easy, safe, inexpensive, and effective analgesia for transnasal fiber-optic laryngoscopy. PMID- 20015728 TI - Complications in pediatric cochlear implants. AB - OBJECTIVE: The purpose of this study is to retrospectively review the complications of pediatric patients undergoing cochlear implantation at a tertiary referral center. METHOD: Institutional review board permission was obtained. A retrospective analysis of all pediatric patients (younger than 18 years) who underwent primary cochlear implantation was performed from January 2001 to December 2005. The patients were reviewed for demographic information, type of hearing loss, cochlear implant device, and complications including implant failure, meningitis, hematoma, implant extrusion, cerebrospinal fluid leak, facial palsy, and wound infection. RESULTS: One hundred sixty-five patient records were reviewed. Twenty-nine patients were lost to follow-up or were revision cases; therefore, 136 records were analyzed. Of the patients, 53.5% were male. The most common etiology of hearing loss was nonsyndromic, nongenetic, congenital sensorineural hearing loss (60.6%). Other less common etiology of hearing loss included TORCH (toxoplasmosis, other infections, rubella, cytomegalovirus, herpes simplex virus) (8.3%), connexin mutation (5.8%), and enlarged vestibular aqueduct (6.5%). All patients had a follow-up of 3 years. There were no intraoperative complications. The most common complications were flap infections (2.6%) and immediate postoperative hematomas (1.9%). Flap problems mostly occurred within 2 weeks of implantation. Within the study period, there was only one device failure (0.7%). CONCLUSIONS: Cochlear implantation in children continues to be reliable and safe in experienced hands, with a low percentage of severe complications. The patients should have a lifetime follow up. PMID- 20015729 TI - Endoscopic above and below approach with frontal septotomy in a patient with frontal mucocele: a contralateral bypass drainage procedure through the frontal septum. AB - Frontal sinus has complex anatomy and is the most difficult sinus to dissect under the nasal endoscope. In case of difficult accessibility through the frontal recess, we can make a detour to more invasive and external procedures to treat chronic or intractable frontal sinus diseases. However, these approaches usually need advanced surgical skills and sometimes can result in minor and/or major complications. Therefore, we developed a new surgical technique to treat frontal mucocele in a patient with severe new bone formation at the frontal recess and presented our experiences with literature review. PMID- 20015730 TI - Bacteriology and antimicrobial susceptibility of pediatric chronic rhinosinusitis: a 6-year result of maxillary sinus punctures. AB - PURPOSE: Few studies in the past decade have focused on antimicrobial resistance of bacteria in pediatric rhinosinusitis. This study aimed to characterize organisms cultured from pediatric chronic rhinosinusitis, as well as current resistance patterns of pathogens. MATERIALS AND METHODS: The study was conducted from January 2001 to December 2006. Children with radiograph-proven chronic rhinosinusitis underwent maxillary sinus punctures to obtain pathogens and for analysis of antibiotic resistance. RESULTS: The total 295 cultures obtained from 165 children yielded 399 isolates. The most common isolates were alpha-hemolytic Streptococcus (20.8%), Haemophilus influenzae (19.5%), Streptococcus pneumoniae (14.0%), coagulase-negative Staphylococcus (13.0%), and Staphylococcus aureus (9.3%). Anaerobes accounted for 8.0% of all isolates. Susceptibility rates of H influenzae for ampicillin and co-trimoxazole were 44.7% and 42.1%, respectively, in the first 3 years of the study and 25% and 40%, respectively, in the next 3 years. Susceptibility rates of S pneumoniae were 83.3% for penicillin, 0% for erythromycin, and 33.3% for clindamycin in the first 3 years and 73.7%, 5.3%, and 28.9%, respectively, in the latter 3 years. CONCLUSION: This study showed a different pattern of antibiotic resistance in pediatric chronic rhinosinusitis as compared with previous studies in both children and adults. The resistance rate of H influenzae for ampicillin appears to be a growing problem in pediatric rhinosinusitis. PMID- 20015731 TI - Visual recovery after direct traumatic optic neuropathy. AB - Direct traumatic optic neuropathy is a rare complication of endoscopic sinus surgery and can result in irreversible damage to the optic nerve. We report a case of direct traumatic optic neuropathy after transnasal endoscopic orbital decompression for Graves's disease in a patient who experienced near-complete recovery of vision. We discuss possible mechanisms. PMID- 20015732 TI - A new method for osteotomies in oncologic nasal surgery: Piezosurgery. AB - OBJECTIVE: Piezosurgery is a recently developed system for cutting bone without necrosis and nonmineralized tissues damage. The aim of this work has been to test Piezosurgery as a new bony scalpel in nasal surgery. METHODS: In this nonrandomized study, we have performed Piezosurgery in the excision of malignant nasal tumors through a paralateronasal approach. We have used Piezosurgery on 10 patients affected by nasal adenocarcinoma. The piezoelectric device uses low frequency ultrasonic waves (24.7-29.5 kHz); the applied power can be modulated between 2.8 and 16 W and is programmed in accordance to the density of the bone. The equipment consists of 2 hand pieces, 2 inserts, and 2 peristaltic pumps; the microvibrations that are created in the piezoelectric hand piece cause the inserts to vibrate linearly between 60 and 210 mum. RESULTS: In all the patients, Piezosurgery provided excellent control without bleeding and harming effects on the adjacent structures. No patients experienced adverse effects. CONCLUSION: Piezosurgery is a new and revolutionary osteotomy technique using the microvibrations of scalpels at ultrasonic frequency, so that soft tissue will not be damaged even upon accidental contact with the cutting tip. The safety of Piezosurgery as regards soft tissues was confirmed. No adverse effects were detected during unintentional contact with the tumor, nerve, vessel, and mucoperiosteum; this renders the piezoelectric device ideal for this application. PMID- 20015733 TI - Cochlear dysfunction in hyperuricemia: otoacoustic emission analysis. AB - PURPOSE: The objective of this study is to provide evidence that primary hyperuricemia is associated with cochlear dysfunction as other metabolic diseases that interfere with cell metabolism. MATERIALS AND METHODS: Cochlear function was evaluated in 25 subjects with asymptomatic hyperuricemia using routine diagnostic audiometry along with transient evoked and distortion product otoacoustic emissions (TEOAE and DPOAE, respectively). To support the notion that vascular compromise was a significant underlying factor for such cochlear dysfunction, we assessed vascular anatomical and functional states through measuring the common carotid artery intima-media thickness and flow velocity of the basal intracranial vessels. RESULTS: Compared with control subjects, reduced response levels of TEOAEs (P < .01) and amplitudes of DPOAEs (P < .001) were detected at higher frequencies. The reduced DPOAE levels at 5 kHz and TEOAEs at 4 kHz correlated significantly with uric acid (P < .05; P < .01), patients' age (P < .06; P < .05), duration since diagnosis of hyperuricemia (P < .05; P < .001), common carotid artery intima-media thickness (P < .05), mean flow velocities of middle cerebral arteries (P < .05), and vertebral arteries (P < .01). Multivariate analysis showed that the abnormalities at higher frequencies were significantly correlated with the duration and degree of hyperuricemia. CONCLUSIONS: These data suggest that subclinical changes in cochlear function are associated with hyperuricemia. They support the usefulness of otoacoustic emissions in early detection of cochlear dysfunction. It is possible that hyperuricemia could be accompanied by increased stiffness and/or compromise of blood supply of the outer hair cells, which will impair their electromotile response. PMID- 20015734 TI - Changing trends of peritonsillar abscess. AB - OBJECTIVE: This retrospective, cohort study aims to assess the changing characteristics of peritonsillar abscess (PTA). METHOD: Data were obtained from PTA patient records admitted to a secondary hospital over a 10-year period. RESULTS: A total of 427 patients, aged 31.6 +/- 15.2 years (range, 3-91), were treated for PTA, reflecting an incidence of 0.9/10 000/y. Forty-seven (11%) patients had more than one episode. There was no sex, seasonal, or side predominance. Thirteen (3%) patients developed complications. One hundred four (24.4%) patients were 40 years or older, had a longer hospital stay, and were prone to complications. One hundred two (23.8%) patients did not have an anteceding pharyngotonsillitis. Smoking was more common among patients with PTA as compared with the general population and was associated with more complications. A total of 283 (66.2%) patients developed PTA in spite of prior antibiotic therapy; 51.1% of smokers that received prior antibiotics had a higher incidence of Streptococcus viridans isolates. CONCLUSION: Peritonsillar abscess may have changed its characteristics: affecting more older patients having a worse and longer course and PTA evolvement without anteceding tonsillitis or in spite of a prior adequate antibiotic therapy. Smoking may be a predisposing factor. PMID- 20015735 TI - Different patterns of second primary malignancy in patients with squamous cell carcinoma of larynx and hypopharynx. AB - PURPOSE: The aim of the present study was to compare the incidence, patterns, and survival of second primary malignancy (SPM) in patients with squamous cell carcinoma of the larynx (LSCC) and hypopharynx (HPSCC). METHODS: We retrospectively review the medical record of 581 previously untreated patients with LSCC (392 cases) and HPSCC (189 cases) who received primary surgery with or without postoperative radiotherapy from 1990 to 2000. Data including age, sex, risk factors, subsites and TNM stage of primary tumor, treatment, site and incidence of SPM, and prognosis were collected from medical charts. RESULTS: Groups with HPSCC had a higher incidence (4.2% vs 2.9% annual rate) and shorter median time (30 vs 59 months) developing SPM rather than LSCC. Fifty-five percent of the SPM occurred in the respiratory axis in LSCC, and 66% developed in the digestive axis in HPSCC. The factors influencing the appearance of SPM included early T stage, tobacco use, and less tumor recurrence. Long-term survival was poorer in those with than without SPM (38% vs 49% at 10 years). CONCLUSIONS: There is a tendency for SPM to occur in the respiratory axis (lung and larynx) in LSCC and in the digestive axis (oral cavity, pharynx, and esophagus) in HPSCC. This information is important for posttreatment follow-up. PMID- 20015736 TI - Radiotherapy-induced common carotid pseudoaneurysm presenting with initially occult upper airway hemorrhage and successfully treated by endovascular stent graft. AB - Radiation induced carotid vasculopathy may present as steno-occlusive disease or less commonly as a pseudoaneurysm. The latter most often presents with a pulsatile mass but is a potential cause of life threatening hemorrhage. We present a case of a small common carotid artery (CCA) pseudoaneurysm that was initially dismissed as the cause of the patients presenting epistaxis given its small size and location. After standard bilateral internal maxillary artery embolizations failed to prevent significant subsequent pharyngeal and tracheal blood loss and serial imaging demonstrated a progressive enlargement of the pseudoaneurysm, a stent graft was successfully placed across the lesion. At five months post stenting, follow-up imaging of the neck showed a stable obliteration of the pseudoaneurysm, good arterial patency, and the patient remained free of recurrent hemorrhage. This case demonstrates that even a small carotid pseudoaneurysm, can present with pharyngeal hemorrhage and should be treated aggressively--with endovascular stent grafting being a preferred treatment modality for arterial lesions in the irradiated neck. PMID- 20015738 TI - Test-retest tinnitus characteristics in patients with noise-induced hearing loss. AB - PURPOSE: The purpose of the study was to examine the test-retest value of tinnitus pitch and loudness in patients with tinnitus and noise-induced hearing loss (NIHL). MATERIALS AND METHODS: The study sample consisted of 30 patients of mean age 35 +/- 6.7 years with long-standing tinnitus and hearing loss due to exposure to noise during military service. Ten patients had unilateral tinnitus, and 20 had bilateral tinnitus. All presented with a typical NIHL audiogram on the affected side(s). None of the patients was receiving drug therapy. RESULTS: There was no statistically significant difference in tinnitus pitch or loudness between the 2 tests for the whole group and separately in patients with unilateral or bilateral tinnitus. CONCLUSION: Subjective testing of pitch and loudness of tinnitus secondary to NIHL is accurate and reproducible, making it a valuable tool for diagnosis and follow-up. The lack of differences between patients with unilateral or bilateral tinnitus indicates that both types may be managed in a similar manner. PMID- 20015737 TI - Operative findings in the frontal recess at time of revision surgery. AB - OBJECTIVE: Endoscopic sinus surgery is the gold standard for the treatment of medically refractory chronic rhinosinusitis. There is, however, a population of patients for whom persistent disease is a problem. Of all the sinuses, the frontal sinus is the most likely to have recurrent obstruction. We evaluated the findings causing frontal recess obstruction at the time of revision surgery. STUDY DESIGN AND SETTING: A retrospective review was performed in a tertiary care academic otolaryngology department. RESULTS: Findings obstructing the frontal recess at the time of revision sinus surgery were reviewed. Two hundred eighty nine frontal sinuses were included. Seven findings were identified: mucosal disease (67%), retained ethmoid cells (53%), lateralized middle turbinates (30%), retained agger nasi cells (13%), scar (12%), retained frontal cells (8%), and neoosteogenesis (7%). Most frontal recesses had multiple etiologies for failure listed above, with an average of 1.6. CONCLUSIONS: Multiple findings can be identified that contribute to frontal recess obstruction requiring revision sinus surgery. A comprehensive approach to address all factors is necessary to prevent surgical failure among patients presenting for endoscopic frontal sinus surgery. PMID- 20015739 TI - Otophyma: a rare and frequently misdiagnosed entity. AB - Otophyma is the term used for sebaceous gland hypertrophy and hyperplasia of the ear. It is usually the end stage of rosacea--a common, chronic, cutaneous disorder of unknown etiology. We hereby present a rare case of otophyma with its clinical features, its etiology, diagnosis, and review of medical and surgical therapy. This patient remained undiagnosed and mismanaged for many years before she was finally diagnosed with this rare condition and successfully managed by surgical technique. To our knowledge, this is the second case of otophyma presented in the otolaryngological literature. PMID- 20015740 TI - Pneumocele of the frontal sinus producing orbital roof defect: case report and review of literature. AB - Type III frontal recess air cell as a cause of frontal sinus pneumocele has not been previously reported in literature. A 31-year-old woman with chronic history of sinusitis presented with pressure in the left eye on blowing the nose. Computed tomography examination of the orbits and paranasal sinuses with coronal and sagittal reformatted images showed abnormal collection of gas in the soft tissues at the superior aspect of the left orbit contiguous with the overlying left frontal sinus through a large defect in the orbital roof and a type III frontal recess air cell narrowing the left frontal recess. PMID- 20015741 TI - Myospherulosis from antibiotic ointment in the postoperative mastoid space. PMID- 20015742 TI - Reconstruction of 2 mandible defects with one fibular flap. PMID- 20015743 TI - Impact of preepiglottic space tumor involvement on concurrent chemoradiation therapy. AB - PURPOSE: The aim of the study was to determine the prognostic impact of preepiglottic space (PES) involvement on local failure after concurrent chemoradiation therapy for squamous cell carcinoma. MATERIALS AND METHODS: Retrospective chart review of patients who underwent concurrent chemoradiation therapy for T3 or T4 laryngeal, T4 hypopharyngeal, and T3 or T4 oropharyngeal squamous cell carcinoma were eligible for inclusion. Patients were then stratified by the presence or absence of PES tumor involvement. A multivariate analysis was performed on the presence of recurrence using the following pretreatment variables: PES involvement, tumor extent, pathologic cell differentiation, lymph node involvement, age, and sex. RESULTS: A total of 102 patients were included in the study. Twenty-seven (28%) patients had documented PES involvement. Mean follow-up for all patients was 46 months. Involvement of the PES was not significantly associated with local tumor persistence or recurrence (P = .69). No other variables significantly impacted tumor recurrence. CONCLUSION: Preepiglottic space involvement does not negatively impact local tumor control after concurrent chemoradiation therapy. PMID- 20015744 TI - Submandibular gland injury and delayed airway compromise caused by a seat belt. AB - Isolated blunt injury to the submandibular gland (SMG) is rare owing to the protection afforded by the overlying mandible. Like other causes of submandibular swelling, glandular trauma can lead to life-threatening airway compromise, and this possibility should be considered in patients presenting with acute pain and fullness in the jaw and neck after trauma. We describe a patient with SMG disruption caused by a seat belt shoulder strap, discuss the diagnosis and treatment, and present a review of the literature. PMID- 20015746 TI - Subcutaneous emphysema and pneumomediastinum after tonsillectomy. AB - Many patients and their families regard tonsillectomy as a minor operation because it is performed so commonly. However, although tonsillectomy is indeed a relatively safe surgical procedure, in extremely rare cases, it can be complicated by subcutaneous emphysema and pneumomediastinum. Although these complications resolve spontaneously in most cases, a few conclude in tension pneumothorax and other fatal complications. The mechanism by which subcutaneous emphysema and pneumomediastinum develop after tonsillectomy is poorly understood. We experienced a case in which subcutaneous emphysema, pneumomediastinum, and a deep defect in the tonsillar fossa were observed. The passage of air was shown by radiology and histopathology. Consequently, the case is reported here. PMID- 20015747 TI - Outcomes of septoplasty in young adults: the Nasal Obstruction Septoplasty Effectiveness study. AB - The goal of this study was to compare the outcome of septoplasty in our patients with previous reports. We found some different outcomes of septoplasty at our center compared with the reports in the literature. One of the major differences between this and previous studies is in the mean age of patients undergoing surgery, 22.44 years in our study vs more than 40 years in most studies. In this study, 86 patients with septal deviation were asked using an outcomes instrument (the Nasal Obstruction Symptom Evaluation scale) before and 3 and 6 months after septoplasty. Seventy-seven patients (89.5%) reported a subjective improvement in their nasal obstruction, which is more than the experience of most authors. There was a significant improvement in mean Nasal Obstruction Symptom Evaluation score at 3 months after septoplasty, and some symptom improvement continued to 6 months. We conclude that younger patients who have nasal obstruction with septal deviation benefit more from septoplasty. PMID- 20015748 TI - Magnetic resonance imaging screening in acoustic neuroma. AB - OBJECTIVES: Magnetic resonance imaging (MRI) is the definitive investigation for detection of an acoustic neuroma. It is however an expensive resource, and pick up rate of a tumor can be as low as 1% of all patients scanned. This study aims to examine referral patterns for MRI screening for patients presenting with asymmetrical sensorineural hearing loss (ASHL). A second aim was to suggest appropriate screening criteria. METHOD: All 132 MRI scans performed for ASHL in the year 2005 were reviewed retrospectively along with their case records and audiograms. In addition, MRI scans and case records were reviewed for the last 30 patients diagnosed with acoustic neuromas. Information was analyzed using 2 published protocols and additional frequency-specific defined criteria. RESULTS: Two acoustic neuromas were picked up out of 132 scans performed. Of the scans performed for ASHL, a third did not fit with any of the published criteria. Of the 30 positive scans for a tumor, the patients/audiograms revealed that 10% did not fit the published criteria despite the patients having no other audiovestibular symptoms. CONCLUSIONS: There appears to be no universally accepted guidelines on screening in ASHL with clinical acumen being used by most ENT consultants in this region. Applying protocols may reduce the amount of scans performed, but up to 10% of tumors may be missed by this approach. PMID- 20015749 TI - Polyarteritis nodosa: a human temporal bone study. AB - PURPOSE: Polyarteritis nodosa is one of the common forms of vasculitis with multiorgan involvement. Hearing loss may be the presenting symptom of this disease. The aim of this histopathologic study was to evaluate temporal bone changes in polyarteritis nodosa and assess the correlation between otologic manifestations and histopathologic findings. MATERIALS AND METHODS: A retrospective human temporal bone analysis was performed at an otopathology laboratory in a tertiary academic medical center. Three cases (5 temporal bones) with pathologically confirmed diagnosis of polyarteritis nodosa were selected for study. The processed temporal bone sections were studied under light microscopy. RESULTS: Two of the 3 cases presented with hearing loss, and one of them had facial palsy. Audiometric data available for one patient demonstrated a bilateral, rapidly progressive, mixed hearing loss. Histopathologically, we found generalized vasculitis involving small- and medium-sized arteries in all cases. Of the 5 temporal bones available for study, 3 had chronic otitis media and 2 had serous otitis media. Arteritis was seen in all middle ears. There was disruption of the organ of Corti and spiral ganglion cells in 4 temporal bones. One temporal bone showed fibrosis and osteogenesis in the scala tympani of the basal turn. Vasculitis of stylomastoid and branches of labyrinthine artery were also found in temporal bones. CONCLUSIONS: Significant temporal bone findings were seen in patients with polyarteritis nodosa, including otitis media, cochlear damage, neo osteogenesis, and middle and inner ear vasculitis. This temporal bone series also suggests an interesting relationship between sensorineural hearing loss and labyrinthine vasculitis. PMID- 20015750 TI - Treatment of pneumocephalus after endoscopic sinus and microscopic skull base surgery. AB - BACKGROUND: Pneumocephalus is a rare complication of endoscopic sinus surgery (ESS) and microscopic skull base surgery (MSBS). Postoperatively, patients often present with headache and altered mental status. Unrepaired leaks are associated with an increased risk of ascending meningitis. Standard treatment of pneumocephalus after ESS or MSBS has not been addressed in the literature. METHODS: The study involved a retrospective review of patients at an academic tertiary care center with pneumocephalus after ESS or MSBS. RESULTS: Ten cases of pneumocephalus were identified, 8 after ESS and 2 after MSBS. Seven ESS defects were very small (<3 mm). The remaining three had defects more than 1 cm. Six of the 8 ESS patients had spontaneous resolution of their cerebrospinal fluid (CSF) leak and pneumocephalus, whereas all patients with larger defects failed conservative therapy with lumbar drainage. Lumbar drainage worsened the pneumocephalus in the MSBS patients. Despite resolution of pneumocephalus in many patients, all were recommended to undergo endoscopic exploration due to concern for increased risk of meningitis in unrepaired defects. Endoscopic repair was necessary in 8 cases. Repairs required a mucosal graft with or without a bone graft. Many spontaneously healed defects were found to be very tenuous. In 2 cases, patients were found to have dense scar over the defect not requiring repair. CONCLUSIONS: In patients with pneumocephalus after ESS or MSBS, consideration should be given to endoscopic exploration and repair of the defect with mucosal grafting. Even if spontaneous resolution has occurred, there may be increased risk of ascending meningitis through the thin or incompletely regenerated mucosa. PMID- 20015751 TI - Hidden unilateral agenesis of the frontal sinus: human cadaver study of a potential surgical pitfall. AB - PURPOSE: The aim of the study was to attract attention to the surgical significance of unilateral agenesis of the frontal sinus hidden by the overlapping expansion of the contralateral sinus toward the agenetic side. MATERIALS AND METHODS: Retrospective review of endoscopic transnasal sinus dissections of 55 human cadavers (42, formalin fixated; 13, fresh frozen) was done in a tertiary care academic medical center. Surgical and radiologic findings were noted. RESULTS: Absence of right frontal sinus ostium in the presence of a connection between the right and left frontal sinuses was demonstrated in 2 (3.6%) cadavers. An absent and an incomplete septum between the frontal sinuses were also noted in these cadavers. No accompanying abnormality of other sinuses was found, and no evidence of previous sinus surgery was noted in these 2 cadavers. CONCLUSIONS: If one of the frontal sinus ostia cannot be found during sinus surgery, although this sinus and its recess can be seen on the thick-sliced coronal computed tomographic (CT) scans, keep in mind that it may be (3.6%) an agenetic frontal sinus hidden by the extensive pneumatization of the contralateral sinus that is crossing the midline. It may not be possible to foresee this variant preoperatively by endoscopic examinations or thick-sliced CT scans. If there is suspicion, thin-sliced CT scans with reconstruction will be ideal to confirm the agenesis of the frontal sinus and to avoid complications. In the presence of such variant of frontal sinus, 1-sided successful frontal sinusotomy is adequate because this sinus or cell will already be drained through the treated frontal recess. PMID- 20015752 TI - Malleus handle: determinant of success in ossiculoplasty. AB - OBJECTIVE: The purpose of this study is to identify prognostic factors affecting outcome in ossicular chain reconstruction (OCR). STUDY DESIGN AND SETTING: This study is a retrospective case series of electronic database at an academic institution. MATERIALS AND METHODS: We reviewed 209 cases of chronic supportive otitis media performed from January 2000 through December 2007 and collected demographic, clinical, audiologic, and outcome information. Univariate analyses of group differences in terms of postoperative air-bone gap (ABG) changes were evaluated by analysis of variance. Multiple regression analyses were used to examine the relationship between postoperative ABG and the independent variables. RESULTS: There were 105 cases of OCR the met the inclusion criteria (44 primary and 61 revision tympanoplasties), with an average follow-up of 19 months. The diagnoses were chronic suppurative otitis media without cholesteatoma in 36 cases and cholesteatoma in 69 cases. The mean preoperative ABG was 34 +/- 15 dB, and the mean postoperative ABG was 20 +/- 14 dB (P < 0.001). Of the independent variables analyzed, the type of procedure (ie, OCR performed during second-look tympanoplasty vs canal wall up vs canal wall down), preoperative ABG, and status of malleus handle were predictive of the success of OCR. CONCLUSIONS: Favorable prognostic factors in OCR include smaller preoperative ABG and the presence of an intact malleus handle. PMID- 20015753 TI - Biofilms and chronic otitis media: an initial exploration into the role of biofilms in the pathogenesis of chronic otitis media. AB - PURPOSE: The aim of the study was to compare the extent of biofilm infection in percentage of mucosal surface area of adenoids removed from children with otitis media with effusion (OME) vs those with recurrent acute otitis media (RAOM) and obstructive sleep apnea (OSA). MATERIALS AND METHODS: Comparative microanatomical investigation of adenoid mucosa using scanning electron microscopy obtained from 30 children with OME, RAOM, and OSA was used in this study. Seventeen males and 13 females ranging in age from 9 months to 10 years were included in this study. Percentage of biofilm surface area involvement was the main measure. RESULTS: Adenoids removed from patients with OME had moderately dense mature biofilms covering the mucosal surface with a mean of 27.7% of their mucosal surface covered with mature biofilms. These results were distinct from results obtained from patients diagnosed with RAOM and OSA with means of 97.6% and 0.10% of their mucosal surfaces covered with mature biofilms, respectively. These differences were statistically significant at P < .0001. CONCLUSIONS: Adenoids removed from patients with OME were characterized by distinctly different percentage of biofilm mucosal surface area coverage, with significantly more biofilm presence than OSA patients but significantly less biofilm presence than RAOM patients. Although previous investigations have supported a dominant role of nasopharyngeal biofilms in RAOM pathogenesis, these results suggest nasopharyngeal biofilms may play a different role in the pathogenesis of OME and that this clinical entity may be more multifactorial in nature. PMID- 20015754 TI - The oncologic safety and functional preservation of supraglottic partial laryngectomy. AB - OBJECTIVE: The purpose of this study was to evaluate the appropriateness of the conservative surgery as the initial treatment of supraglottic cancer by the comparative study of supraglottic partial laryngectomy with radiation therapy. METHODS: We analyzed 48 patients diagnosed as having supraglottic cancer who underwent supraglottic partial laryngectomy and 20 patients treated with radiation therapy as initial treatment from 1991 to 2005. Oncologic safety was evaluated by comparison of survival rates. Swallowing test was performed with videoesophagofluoroscopy under the category of the pharyngeal phase and the presence or absence of aspiration. Phonologic analysis was performed with acoustic waveform analysis and stroboscopy. RESULTS: Positive resection margin was observed in 12 of the patients who received supraglottic partial laryngectomy (SPL), and its relationship with the primary subsite was evaluated. For the patients in stages I and II, the overall survival rate in each of the SPL group and the radiotherapy (RTx) group was 87% and 80%, respectively. For the patients in stages III and IV, the overall survival rate was 87% and 80%, respectively. On evaluation of swallowing and phonation function, no significant difference was noted between the SPL group and the RTx group. CONCLUSION: This study suggests that for patients with supraglottic cancer, SPL provides acceptable survival, swallowing, and phonologic results in comparison with radiation therapy. PMID- 20015755 TI - Spontaneous tracheal rupture caused by vomiting. AB - OBJECTIVES: To present a pediatric case of spontaneous tracheal rupture caused solely by vomiting and to discuss its diagnosis and management. STUDY DESIGN: Case presentation and literature review. RESULTS: A 14-year-old girl with a new diagnosis of type 1 diabetes mellitus presented with respiratory distress. History was significant only for 4 days of violent vomiting, and she was diagnosed with diabetic ketoacidosis. Examination revealed tachypnea and considerable subcutaneous air overlying the upper chest and neck; chest x-ray showed pneumomediastinum. A swallow study showed no evidence of an esophageal tear. Computed tomography of the chest showed a posterior tracheal tear 4.5 cm distal to the cricoid cartilage. The patient's ketoacidosis was controlled, and supplemental oxygen was administered temporarily to promote absorption of the extravasated air. Serial chest x-rays showed complete resolution within 5 days and intubation was not required. CONCLUSIONS: To our knowledge, this is the first reported pediatric case of tracheal rupture secondary to vomiting. Previous reports of spontaneous tracheal rupture have been related to paroxysmal coughing. In similar cases of subcutaneous emphysema and pneumomediastinum after retching, an esophageal rupture should be ruled out. Conservative management was successful in this case. PMID- 20015756 TI - Atypical Cogan's syndrome: a case report. AB - Cogan's syndrome is a rare presumed autoimmune disorder characterized by nonsyphilitic interstitial keratitis and progressive audiovestibular symptoms. The initial report by David G. Cogan in 1945 was modified by Haynes et al in 1980 who proposed diagnostic criteria for patients with other ocular or vestibular symptoms and suggested this to be atypical Cogan's syndrome. In a more typical presentation of Cogan's syndrome, ocular and audiovestibular signs and symptoms usually appear alone and are bilateral. We report a case of 50-year-old woman with an atypical Cogan's syndrome manifested by unusual relatively rapid clinical deterioration. PMID- 20015757 TI - The use of complementary and alternative medicine by patients attending a general otolaryngology clinic: can we afford to ignore it? AB - BACKGROUND: There is growing interest in use of complementary and alternative medicine (CAM) among the general population. Little information is available, however, on CAM use in adults attending an otolaryngology outpatient clinic in the UK. OBJECTIVE: The purpose of this article is to study the prevalence and pattern of CAM use among adult patients attending the ear, nose, and throat (ENT) clinic in a UK teaching hospital. STUDY DESIGN: A cross-sectional study was used. SUBJECTS AND METHODS: All patients on their primary visit to an ENT clinic were asked to complete an anonymous questionnaire for a 14-week period from October 2005 to January 2006. RESULTS: Based on 1366 completed questionnaires, 53% (728/1366) were female and 47% (638/1366) were male. Twenty-nine percent (395/1366) were older than 60 years, and 56% (763/1366) were married. Twenty percent (275/1366) had a university education. Sixty-one percent (833/1366) had used CAM, almost 36% in the preceding 12 months. The popular remedies were cod liver oil (368/833), garlic (197/833), cranberry (181/833), aloe vera (176/833), primrose oil (174/833), and Echinacea (163/833). Nonherbal therapies included massage (230/833), acupuncture (186/833), aromatherapy (135/833), chiropractic (121/833), reflexology (119/833), and homeopathy (110/833). Seventeen percent (143/833) used CAM for their current illness. Only 8% (64/833) found CAM ineffective; yet, 57% (473/833) would recommend CAM to others. Fifty-one percent (421/833) failed to inform their primary physician of their CAM use. CONCLUSION: Despite concerns over CAM efficacy, safety, and cost effectiveness, use of CAM is popular among patients attending an ENT clinic. Their use is not generally related to their presenting illness. Otolaryngologists should be aware of current trends in CAM use when managing patients, including possible interactions with other medication. PMID- 20015758 TI - Carotid artery blow-out during mastoidectomy: management using endovascular coils. PMID- 20015759 TI - Endoscopic management of a massive nasal osteoma presenting as an orbital abscess. PMID- 20015760 TI - Endoscopic treatment of tracheocele in pediatric patients. AB - Acquired tracheal pouch known also as tracheocele is a rare air-filled diverticulum of tracheal pars membranacea. This disease may be due to esophageal or tracheal surgery, orotracheal intubation, or increased intralunimal pressure through a weak area of tracheal wall. When symptomatic and medical therapy is insufficient, this disorder must be treated surgically. Several surgical methods ranging from open neck or thoracic surgery to endoscopic managements have been reported. We report the case history of a 7-year-old boy affected by recurrent pneumonia due to tracheal pouch. The patient underwent successful brushing of tracheocele plus fibrin glue application by rigid tracheobroncoscopy. Furthermore, particular emphasis on endoscopic treatments of tracheal diverticulum is also presented. PMID- 20015761 TI - Toxic epidermal necrolysis caused by cetuximab plus minocycline in head and neck cancer. AB - We present a unique case of toxic epidermal necrolysis (TEN) caused by cetuximab plus minocycline during chemoradiation for recurrent squamous cell carcinoma of the head and neck (SCCHN). In April 2008, a 49-year-old man was diagnosed with recurrent SCCHN. He received surgery and postoperative adjuvant therapy with cetuximab plus chemoradiation. Because of exacerbation of acneform rash by cetuximab, minocycline was added thereafter. However, TEN appeared after the fifth course of weekly cetuximab, and cetuximab plus minocycline was assumed to be the cause. Cetuximab plus minocycline carries a risk of evoking TEN, and the side effects of chemoradiation for SCCHN mask some early signs of TEN. Noticing the transformation of skin lesions makes early diagnosis possible, and prompt cessation of the causative agent is essential for patients with TEN. We review the literature on cetuximab, minocycline, and TEN and discuss the impact of TEN on cancer therapy. PMID- 20015763 TI - Imaging findings in a case with cholesteatoma in complete aural atresia. AB - We report a case of cholesteatoma in congenital aural atresia in which preoperative imaging analysis provided useful information. A 30-year-old man with total aural atresia had had ear symptoms on his right side for 10 years. Evaluation by computerized tomography and magnetic resonance imaging suggested that the mass in the atretic external auditory canal and the anterior part of the mastoid was cholesteatoma and that the mass in the posterior part of the mastoid was inflammatory granulation tissue and chronic osteomyelitis. Three-dimensional fluid-attenuated inversion recovery images showed increased signals in the labyrinth. The extent of the cholesteatoma revealed during surgery was consistent with the preoperative image findings. Imaging analysis provides useful information for management of congenital aural atresia, and combined examination with computerized tomography and magnetic resonance imaging could be beneficial for preoperative evaluation of state of ears in cases of congenital aural atresia. PMID- 20015762 TI - Malignant transformation of a highly aggressive human papillomavirus type 11 associated recurrent respiratory papillomatosis. AB - OBJECTIVE: The objective is to present an uncommon case of squamous cell carcinoma (SCC) arising from extensive recurrent respiratory papillomatosis (RRP) involving the upper and lower airway and temporal bone. STUDY DESIGN: This is a case report and a review of the literature. METHODS: We describe a case of a 24 year-old woman with a history of human papillomavirus (HPV) type 11 since childhood originating in the larynx and trachea, then progressing to involve the distal pulmonary alveoli and right middle ear through the eustachian tube. Papillomatous growth was treated with multiple surgeries including laser cytoreduction of laryngotracheal papillomatosis and radical mastoidectomy, followed by a trial of chemotherapy. Despite this aggressive treatment regimen, papillomatous growth progressed with recurrence in the right eustachian tube, middle ear, and mastoid eventually extending to involve the calvaria and scalp. RESULTS: The patient underwent a composite resection of involved tissues, including the scalp, auricle, and lateral temporal bone, with reconstruction using a latissimus dorsi free flap. Final pathologic analysis revealed an extensive infiltrative well-differentiated SCC arising from the papilloma. A review of the literature on aggressive respiratory papillomatosis suggests that malignant transformation of juvenile-onset RRP occurs exclusively in cases positive for HPV-11. CONCLUSIONS: We report an unusual case of SCC originating from extensive RRP involving the airway, temporal bone, and scalp and describe the medical and surgical management. Although the incidence of juvenile-onset RRP transformation to SCC is very low, the presence of HPV-11 as a risk factor for malignant transformation of RRP is becoming evident. PMID- 20015764 TI - Sinonasal teratocarcinosarcoma. AB - Sinonasal teratocarcinosarcoma (SNTCS) is a rare and aggressive neoplasm characterized by the combination of malignant teratoma and carcinosarcoma. Sinonasal teratocarcinosarcoma usually is found mainly among adults, and the common original sites of SNTCS were nasal cavities and paranasal sinuses. There are fewer than 50 reported SNTCS cases in the literature. The most common cause of treatment failure is local recurrence. Distant metastasis of SNTCS is seldom described because of the invasive character of the cancer. We report on a 52-year old man with SNTCS involving the left sinonasal cavity. Recurrent cervical metastasis developed in the 18th, 23rd, and 28th month after initial sinonasal surgery. Despite multiple aggressive neck surgeries and postoperative adjuvant radiotherapy, the patient died of lung metastasis 43 months after the initial surgery. Our patient was younger than other reported patients with SNTCS; besides, good locoregional control and lung metastasis have not been reported previously. An aggressive elective neck dissection should be performed in the early disease stage, and more attention should be given to the soft tissue surrounding any possible lymphadenopathy. This may decrease the risk of lower cervical lymph node or distant metastasis in patients with SNTCS. PMID- 20015765 TI - Deep neck abscess due to Acinetobacter baumannii infection. AB - Acinetobacter baumannii strains are isolated in up to 1% of nosocomial infections mostly from intensive care units immunocompromised patients and are associated with high mortality rates. A baumannii infections include pneumonia, urinary tract infection, endocarditis, skin and soft-tissue infections, surgical-site infection, meningitis, osteomyelitis, and septicemia. We report an extremely rare case of deep neck abscess due to multidrug-resistant A baumannii infection. The isolate strain was analyzed by a repetitive sequence-based polymerase chain reaction typing method: the isolate profile was compared with other strains obtained from isolates recovered in the hospital in that period. Our patient underwent 2 neck explorations and antibiotic treatment (tigecycline 50 mg, twice per day). Five weeks after admission, the patient was discharged in good general conditions. Considering the other obtained strains, 4 different profiles were identified, one as prominent (profile A, 18 isolates), the index case (B), and 2 others (C, D) as divergent. PMID- 20015766 TI - Influence of audit on clinical practice: multidisciplinary team data documentation for cutaneous head and neck malignancy. AB - BACKGROUND: Prognosis of cutaneous head and neck malignant neoplasms is related to type, site, and histologic feature but may also be influenced by delays in the referral process. Treatment aims to address the primary lesion and when necessary the secondary disease but at the same time maintaining quality of life and minimizing cosmetic deformity. The management plan should ideally be the "product" of a multidisciplinary team (MDT) approach of dermatologists, surgeons, oncologists, pathologists, and radiologists. AIMS: The purposes of the study were as follows: (i) to review current documentation practice of cutaneous head and neck malignancies in a district hospital, (ii) to generate an MDT-approved data collection proforma, and finally (iii) to reaudit documentation of head and neck skin lesions based on the "new" standardized proforma. The MDT-approved proforma was instituted in an effort not only to collect data in a coherent and structured way but also to achieve early diagnosis and avoid delays from urgent referral to treatment with the aim of curing disease and restoring patients to as near-normal appearance and quality of life as possible. METHODS: The notes of patients with head and neck cutaneous lesions were prospectively audited for a 6-month period (August 2006-January 2007). The presence and absence of a minimal standardized data set required for MDT discussion and review were recorded. RESULTS: Documentation was deficient and a need for change in practice emerged. An MDT approved data collection standardized proforma was designed and distributed throughout the hospital departments involved in management of patients with head and neck cutaneous lesions. The notes of these patients were prospectively reaudited through a second cycle for a 6-month period (February 2007-August 2007). The completion and deficiency of sections of the MDT-approved data collection standardized proforma were recorded, resulting in closure of the audit loop. CONCLUSIONS: The data documentation for head and neck cutaneous lesions has significantly improved after the implementation of an MDT-approved data collection standardized proforma. This has resulted in improved patient quality of care and progression on appropriate management plans. PMID- 20015767 TI - Bone morphogenetic protein-2/4 and bone morphogenetic protein receptor type IA expression in metastatic and nonmetastatic oral squamous cell carcinoma. AB - PURPOSE: The study aimed to analyze the expression of bone morphogenetic protein 2/4 (BMP-2/4) and its receptor BMPR-IA (BMP receptor type IA) in metastatic and nonmetastatic oral squamous cell carcinoma (OSCC) and its implications for disease prognosis. MATERIALS AND METHODS: The experimental group included 16 cases of OSCC without metastasis and 7 cases of OSCC with metastasis. The presence or absence of nodal metastasis was used as a parameter for the evaluation of disease prognosis. Ten cases of oral fibroepithelial hyperplasia were selected as the control group. The expression of BMP-2/4 and BMPR-IA was analyzed by immunohistochemistry. RESULTS: In the experimental group with metastasis, strong expression of BMP-2/4 was observed in most cases (71.4%), whereas BMPR-IA exhibited weak expression (85.7%). In the experimental group without metastasis, there was strong expression of BMP-2/4 (62.5%) and BMPR-IA (100%). A significant association was observed between the prognosis of OSCC and the intensity of BMP-2/4 staining (P = .002). Weak immunoreactivity to BMP-2/4 and BMPR-IA was observed in all control specimens. CONCLUSIONS: The results suggest that strong expression of BMP-2/4, associated with low expression of BMPR IA, observed in metastatic OSCC has a prognostic value, with the loss of responsiveness to BMPs through the loss of expression of their receptors being indicative of the development of metastasis. PMID- 20015768 TI - The importance of vascular endothelial growth factor in the healing of acute tympanic membrane perforation. AB - PURPOSE: The purpose of this study was to determine the more important growth factor expression between basic fibroblast growth factor (bFGF) and vascular endothelial growth factor (VEGF) in the healing of acute tympanic membrane (TM) perforation. MATERIALS AND METHODS: Bilateral perforations of the TM were created in 12 rats. The TM perforations in the right ears were treated with dexamethasone, and left ears were designated as the control group. The TM was examined for the growth factor expression immunohistochemically in the epithelial and fibrous layers according to the rate of TM perforation healing. RESULTS: The mean spontaneous healing time of the TM perforations was 11.0 +/- 2.0 days. However, dexamethasone-treated group showed no evidence of closure. The bFGF and VEGF expression were significantly correlated with the rate of healing of acute TM perforations. The VEGF expression was decreased both in the epithelial and fibrous layers, but bFGF expression was decreased only in the epithelial layer in the dexamethasone-treated group. The VEGF was expressed to a lesser degree than bFGF in the dexamethasone-treated group. CONCLUSION: Vascular endothelial growth factor is the more specific and important growth factor than bFGF in the healing of acute TM perforation. PMID- 20015769 TI - Recurrent contralateral abducens nerve palsy in acute unilateral sphenoiditis. AB - OBJECTIVE: We describe a patient who presented with recurrent, contralateral abducens nerve palsy resulting from acute sphenoiditis. METHODS: Patient medical record and imaging studies were reviewed. A literature review of complications of sphenoiditis was performed. RESULTS: Our patient presented with symptoms consistent with an upper respiratory tract infection, as well as severe headache, disequilibrium, and diplopia. Physical examination demonstrated left-sided sixth cranial nerve palsy. Imaging studies were consistent with right sphenoid sinus inflammatory disease. Her medical history was significant for a similar episode 9 years previously, which was managed medically. The current episode was treated acutely with antibiotics and prednisone. Upon resolution of her symptoms, she underwent an endoscopic sphenoidotomy. CONCLUSIONS: Although cranial nerve palsies associated with isolated sphenoiditis have been reported, we describe a unique case of recurrent isolated sphenoiditis causing contralateral abducens nerve palsy. Acute management can be medical, with surgical therapy reserved for refractory cases or to prevent future episodes. PMID- 20015770 TI - Prognostic value of metastatic lymph node ratio in node-positive head and neck carcinomas. AB - PURPOSE: The purpose of the study was to determine the prognostic significance of the ratio between metastatic and examined lymph nodes to the survival rate of patients with squamous cell carcinoma of the head and neck. MATERIALS AND METHODS: The study included 142 patients in whom metastatic lymph nodes were observed in neck dissection specimens. The number of metastatic lymph nodes and the ratio of metastatic lymph nodes (RMLN) were compared as prognostic factors affecting overall survival (OS) and disease-free survival (DFS) rates. Survival curves were made using Kaplan-Meier analysis and were assessed by the log-rank test and Cox regression method. RESULTS: The median number of metastatic lymph nodes and RMLN for the entire study population were 2 and 4%, respectively. The OS and DFS rates were analyzed in the groups according to such stratification. The result of analysis of OS and DFS showed a statistically significant difference between patients with RMLN of at least 4% and those with RMLN less than 4% (hazard ratio, 3.4 and 2.7; P = .015 and P = .001, respectively). CONCLUSIONS: The ratio of metastatic lymph nodes has a significant impact on the survival period. PMID- 20015771 TI - Foreign body aspiration in children: the effects of delayed diagnosis. AB - BACKGROUND: Foreign body aspiration is common in children, but there are no clinical clues that can determine or rule out the diagnosis before entering the operating room for bronchoscopy. The purposes of our study were to define the clinical characteristics of foreign body aspiration in the pediatric population and to evaluate the significance of delay in its diagnosis and treatment. METHODS AND RESULTS: The study used a retrospective review of 136 charts of children up to the age of 16 who underwent bronchoscopy for foreign body removal for 10 years in Ha'Emek Medical Center (Afula, Israel). An adult witness of an aspiration episode (most frequent presenting symptom), a child younger than 2 years, and an abnormal plain chest radiography were found to be significant predictors of foreign body aspiration. Estimated risk for foreign body aspiration was significantly higher in children who were younger than 2 years with an eyewitness for the aspiration episode and abnormal chest radiography (odds ratio, 5.6, with confidence interval from 2.0 to 15.6). The rate of complication was 2-fold higher in patients who arrived at the hospital 2 days or more after the aspiration compared with patients who arrived earlier. The rate of complication was 2-fold higher in patients who underwent bronchoscopy 24 hours or more after arrival at the emergency department compared with patients who underwent bronchoscopy within the first 24 hours. CONCLUSION: Delayed arrival of a child with a suspected foreign body aspiration at the hospital and delayed bronchoscopy were found to be related to a higher rate of complication. PMID- 20015772 TI - Type 2 ossiculoplasty: prognostic determination of hearing results by middle ear risk index. AB - PURPOSE: The aims of this study were to investigate the prognostic impact of middle ear risk index on the postoperative hearing results in cases with type 2 ossiculoplasty; to compare the middle ear risk index results among primary, staged, and revision cases; and to compare the results of the prostheses used in ossicular reconstruction. MATERIAL AND METHODS: Records of 293 patients who had canal wall up tympanomasteidectomy and type 2 ossiculoplasty due to chronic otitis media between November 1995 and November 2007 were reviewed retrospectively. RESULTS: The mean preoperative air-bone gap was 32.6 dB, and it decreased to 15.2 dB after a mean follow-up period of 26.8 months postoperatively. The mean change of air-bone gap was 17.4 dB. Postoperative air bone gap was 20 dB or less in 79% of the cases. The patients with dry perforations were in the low-risk group, and 91% of them had an air-bone gap of 20 dB or less. This value was 86% in the ones with intact malleus. The patients who had primary surgery were found in moderate risk group, whereas staged and revision groups were in the high-risk group. The air-bone gap was 20 dB or less in 84%, 78%, and 59%, respectively, of those groups. The difference between the primary and the revision groups reached a statistical significance. CONCLUSIONS: We had the best ossicular reconstruction results with glass ionomer cement, whereas the worst results were obtained with allograft partial ossicular replacement prostheses. We determined that risk-reducing factors such as dry ear, minimal ossicular chain defect, and intact malleus were important to have successful results. The middle ear risk index is a valuable tool for the surgeon to judge the risks and the probability success of the procedure as well as to make a good patient selection. PMID- 20015773 TI - Skull base infection presenting with multiple lower cranial nerve palsies. AB - Diseases of the temporal bone causing lower cranial nerve palsies are uncommon. In the presence of bony erosion, they are highly suggestive of a malignant process. However, when there is a clear history of otitis externa in an immunocompromised or diabetic patient, a diagnosis of osteomyelitis and secondary inflammatory mass should be considered. We report 4 separate cases of infective skull base lesions causing multiple lower cranial nerve palsies in elderly patients who were not immunocompromised or diabetic, highlighting that this condition is not exclusive to this population. PMID- 20015774 TI - Benefit from bimodal hearing in a group of prelingually deafened adult cochlear implant users. AB - PURPOSE: People who receive a unilateral cochlear implant find speech perception in acoustically complex situations very challenging, in part because they do not have access to binaural hearing. For cochlear implant patients with some residual hearing in the nonimplanted ear, bilateral auditory input can be obtained by the use of a cochlear implant and a contralateral conventional hearing aid. This condition is referred to as "bimodal hearing." MATERIALS AND METHODS: We evaluated the benefits from bimodal stimulation in a group of 10 prelingually deafened adult cochlear implant users, submitted to unilateral cochlear implantation at the ENT Unit of the University of Pisa. RESULTS: Of 10 patients, 9 decided to continue using bimodal stimulation and demonstrated improvements in speech perception both in quiet and in noise condition from bimodal hearing, in comparison to the cochlear implant alone condition. CONCLUSIONS: Our results show that bimodal hearing offers some advantages in speech understanding in quiet and noise conditions also in prelingually deafened adults. Moreover, most of our patients reported advantages from bimodal hearing in localizing sound and in perceiving a more natural sound. PMID- 20015775 TI - Rhinoscleroma: case report. PMID- 20015776 TI - When the bell tolls on Bell's palsy: finding occult malignancy in acute-onset facial paralysis. AB - PURPOSE: This study reports 4 cases of occult parotid malignancy presenting with sudden-onset facial paralysis to demonstrate that failure to regain tone 6 months after onset distinguishes these patients from Bell's palsy patients with delayed recovery and to propose a diagnostic algorithm for this subset of patients. MATERIALS AND METHODS: A case series of 4 patients with occult parotid malignancies presenting with acute-onset unilateral facial paralysis is reported. RESULTS: Initial imaging on all 4 patients did not demonstrate a parotid mass. Diagnostic delays ranged from 7 to 36 months from time of onset of facial paralysis to time of diagnosis of parotid malignancy. Additional physical examination findings, especially failure to regain tone, as well as properly protocolled radiologic studies reviewed with dedicated head and neck radiologists, were helpful in arriving at the diagnosis. CONCLUSION: An algorithm to minimize diagnostic delays in this subset of acute facial paralysis patients is presented. Careful attention to facial tone, in addition to movement, is important in the diagnostic evaluation of acute-onset facial paralysis. PMID- 20015777 TI - Hemangiomas of the external auditory canal. AB - Hemangiomas of the external auditory canal are rare with only 16 cases reported in the literature. We report the case of a 62-year-old woman who presented with bleeding from the external auditory canal. Physical examination and imaging were consistent with a hemangioma. Initial diagnostic evaluation failed to reveal the full extent of the lesion resulting in recurrence after primary local excision. After recurrence, the hemangioma was managed successfully with embolization and a more extensive resection. In addition, we review the relevant literature with respect to epidemiology, presentation, evaluation, pathology, and management options for hemangiomas arising in the external auditory canal. PMID- 20015778 TI - Endoscopic approach for excision of juvenile nasopharyngeal angiofibroma: complications and outcomes. AB - The purposes of this study are to report the efficacy of the endoscopic approach for juvenile nasopharyngeal angiofibroma (JNA) and to compare its related intra- and postoperative complications with findings from traditional approaches in the literatures. This study is a retrospective report of 47 cases of JNA that were treated with nasal endoscopic surgery between 1998 and 2005. According to the staging system by Radkowski et al (Arch Otolaryngol Head Neck Surg. 1996;122:122 129), the staging of the included patients were the following: 21 in stages IA to IIB, 22 in IIC, 3 in IIIA, and 1 in IIIB. Five patients were embolized before surgery. The mean blood hemorrhage in embolized patients was 770 mL, whereas in nonembolized patients, it was 1403.6 mL. In the follow-up period (mean, 2.5 years), the recurrence was found in 9 patients (19.1%), and mean time of recurrence was 17 months after surgery. The rupture of cavernous sinus occurred in 2 cases with no mortality. The mean hospital stay was 3.1 days in all cases and 1.8 days in embolized patients. The findings of this study demonstrate that endoscopic resection of JNA is a safe and effective technique because of decrease in blood loss, hospitalization, and recurrence rate, especially in tumors that are not extended through intracranial space. It is therefore strongly recommended that this modality is implemented as the first surgical step for tumors with stages I to IIIA of the Radkowski's staging system. PMID- 20015779 TI - Cochlear implantation in patients with bilateral cochlear trauma. AB - PURPOSE: Temporal bone fracture, which involves the otic capsule, can lead to complete loss of auditory and vestibular functions, whereas the patients without fractures may experience profound sensorineural hearing loss due to cochlear concussion. Cochlear implant is indicated in profound sensorineural hearing loss due to cochlear trauma but who still have an intact auditory nerve. MATERIAL AND METHODS: This is a retrospective review study. We report 5 cases of postlingually deafened patients caused by cochlear trauma, who underwent cochlear implantation. Preoperative and postoperative hearing performance will be presented. These patients are cochlear implanted after the cochlear trauma in our department between 2001 and 2006. RESULTS: All patients performed very well with their implants, obtained open-set speech understanding. They all became good telephone users after implantation. Their performance in speech understanding was comparable to standard postlingual adult patients implanted. CONCLUSION: Cochlear implantation is an effective aural rehabilitation in profound sensorineural hearing loss caused by temporal bone trauma. Preoperative temporal bone computed tomography, magnetic resonance imaging, and promontorium stimulation testing are necessary to make decision for the surgery and to determine the side to be implanted. Surgery could be challenging and complicated because of anatomical irregularity. Moreover, fibrosis and partial or total ossification within the cochlea must be expected. PMID- 20015780 TI - Airway implications of congenital sternal agenesis. AB - Congenital agenesis of the sternum is an extremely unusual malformation rarely encountered by the practicing Otolaryngologist. It typically arises in conjunction with other midline ventral congenital anomalies, including abdominal, diaphragmatic, and cardiac malformations. We report a case series of two patients managed with tracheotomy placement due to prolonged intubation. The first patient was a 63-day-old infant born at 34 weeks gestation with dysmorphic features, cleft lip and palate, and skeletal dysplasia, including absence of the sternum. The second patient was a 31-day-old infant born with ectopic cordis and diaphragmatic hernia. The inadequate closure of the anterior chest wall secondary to manubrium malformation or a variant contributes significantly to a child's inability to generate adequate ventilatory pressures. As a result, airway management must be considered not only to ensure airway patency but also an appropriate physiological environment to allow for adequate air exchange in the lungs. PMID- 20015781 TI - Repair of large nasal septal perforation with titanium membrane: report of 10 cases. AB - OBJECTIVE: The aim of the study was to study a new treatment to repair large nasal septal perforation with medical titanium membrane. METHOD: Ten patients with septal perforation underwent repair with open rhinoplasty approach using titanium membrane. RESULTS: The perforation of nasal septum in 10 patients was closed satisfactorily. During our followup period (mean, 1 year), the titanium membrane was mucosalized, and this technique led to persistent closing of perforation. CONCLUSION: Repair of septal perforation with titanium membrane yields excellent results. PMID- 20015782 TI - Epistaxis due to Wegener's granulomatosis in a pediatric patient. AB - Epistaxis is a common problem in children that typically is not severe and seldom requires hospitalization. The nose is a highly vascular structure with a large surface area; subsequently, it is highly predisposed to bleeding. Childhood vasculitides are very rare and are commonly diagnosed by characteristic lesions on imaging studies along with syndrome recognition by clinicians. We present a case of recurrent epistaxis that persisted over 3 months due to Wegener's granulomatosis in an adolescent that was misdiagnosed as a benign hemorrhage from Kiesselbach's plexus. PMID- 20015783 TI - Cochlear implantation in a patient with superficial siderosis: an update. AB - OBJECTIVE: The purpose of this study is to present follow-up on a previously reported case of successful cochlear implantation in a patient with superficial siderosis. STUDY DESIGN: Retrospective case review. RESULTS: For the first 6 years after implantation, the patient had maintained a successful result; however, she developed a progressive decline in the benefit from her implant. CONCLUSION: Benefit from cochlear implants in patients with superficial siderosis is variable and may not be long standing. PMID- 20015784 TI - Radiologic appearance of chronic parapharyngeal Teflon granuloma. AB - Although Teflon has been used for almost 5 decades to provide tissue augmentation in various surgical indications, including head and neck surgery, its use has significantly declined in the last 2 decades, primarily because of its implication in granuloma formation. Teflon granulomas have been shown to cause false positives on positron emission tomography imaging and have been reported to have a characteristic magnetic resonance imaging (MRI) appearance. We report a patient with a large chronic Teflon granuloma of the parapharyngeal space that caused significant bony erosion of the atlas vertebra. The lesion's MRI signal characteristics were indistinguishable from those of surrounding tissues, while it showed characteristic hyperdensity on computed tomography due to the presence of fluorine atoms within Teflon. As MRI may supersede or replace computed tomography for a number of indications, and as Teflon has been used in large numbers of patients whose records may not always be available, knowledge of these findings has clinical relevance. PMID- 20015785 TI - Somatostatin in medical management of chyle fistula after neck dissection for papillary thyroid carcinoma. AB - The use of somastostatin and its analogues in the treatment of chyle fistula is a new approach and has been documented in a few cases. In this study, we present the case of a male patient with chyle fistula that was stopped completely within 24 hours after the somatostatin therapy. PMID- 20015786 TI - "Hide and seek" with antineutrophil cytoplasmic antibodies. PMID- 20015787 TI - Device-related risk of fire in oropharyngeal surgery: a mechanical model. AB - OBJECTIVE: The aim of this study was to evaluate the fire risk for an electrosurgical device (Bovie) as compared with a bipolar radiofrequency ablation wand (Coblator) in a mechanical model of oropharyngeal surgery. METHODS: An endotracheal tube was inserted into the cranial end of a degutted, whole raw chicken through which 100% oxygen was piped at 10 L/min. An electrosurgical device (15 W, coagulate mode) and bipolar radiofrequency ablation wand (9, 7, and 3 in ablate mode; 5 and 3 in coagulate mode) were activated in the central cavity. All experimental conditions were tested for 4 minutes, or until a positive result was achieved. All trials were repeated to ensure accuracy. RESULTS: Ignition was obtained with a sustained fire when using the electrosurgical device for between 25 and 80 seconds. Under all 5 experimental conditions, no ignition or sustained fire could be produced using the radiofrequency ablation wand. After 20 sustained minutes of continuous plasma ablation in the chicken cavity without ignition, the electrosurgical device was able to ignite a fire in 25 seconds in the same cavity. CONCLUSIONS: Although electrosurgical devices present a significant risk of fire during open cavity surgery in oxygen-enriched environments, that risk seems to be eliminated with bipolar radiofrequency plasma ablation. PMID- 20015788 TI - A randomized prospective study of oral levofloxacin vs intravenous flomoxef prophylaxis in postoperative infection after endoscopic sinus surgery. AB - OBJECTIVE: The clinical efficacy and cost effectiveness of oral antimicrobial prophylaxis with levofloxacin (LVFX) on endoscopic sinus surgery (ESS) was evaluated. MATERIALS AND METHODS: Ninety-three patients undergoing ESS were prospectively enrolled in the present study. The patients were randomly divided into 2 groups, LVFX and flomoxef (FMOX). Two hundred milligrams of LVFX was orally given 2 hours before the start of surgery and 6 hours after the end of surgery, which was followed by the administration of 200 mg every 12 hours for 2 days. One gram of FMOX was dissolved in 100 ml of physiological saline and given intravenously at the induction of anesthesia and 6 hours after the end of surgery, followed by infusion twice daily for 2 days. RESULTS: There were no statistically significant differences between 2 groups in terms of age, sex, systemic complications, surgical procedures, the duration of the operation, the length of hospitalization, the amount of blood loss, body temperature, or the number of white blood cells or C-reactive protein. Although no statistical significance was observed in the bacterial resistance between the two antibiotics, LVFX seems to show a low rate of resistance pattern change as compared to FMOX. The present study demonstrated that no patients treated with LVFX or FMOX were afflicted with postsurgical infection. CONCLUSION: Oral administration of LVFX is a simple, cost-effective and safe alternative to intravenous prophylaxis in ESS based on clinical efficacy and bacteriological study. PMID- 20015789 TI - Oscillopsia in labyrinthine defective patients: comparison of objective and subjective measures. AB - OBJECTIVE: To compare the oscillopsia sensation in vestibular defective patients, using a specific handicap questionnaire and a specific Visual Analog Scale, with objective measure of the vertical vestibulo-ocular reflex efficiency in the pitch plane, using the computerized Dynamic Visual Acuity (DVA) test and Gaze Stabilization Test (GST). DESIGN: Controlled retrospective study. SETTING: Day hospital in ENT Rehabilitation Unit. SUBJECTS: Sixty-five subjects: 35 controls (12 men and 23 women; mean age, 50.77 +/- 13.39 years) and 30 patients with chronic dizziness: 18 with unilateral vestibular hypofunction (7 men and 11 women; mean age, 55.50 +/- 12.72 years) and 12 with bilateral hypofunction (7 men and 5 women; mean age, 57.25 +/- 9.18 years). MAIN MEASURES: Computerize vertical DVA and GST; subjective Visual Analog Scale, Oscillopsia Score questionnaire. RESULTS: Instrumental tests had different means between subject groups; vertical DVA results and subjective measures were significantly correlated. CONCLUSIONS: Vertical DVA and GST test in up and down direction are able to separate healthy and vestibular patients. Moreover, the DVA test in down direction differentiates patients with unilateral vestibular hypofunction and with bilateral vestibular hypofunction. These results show that vertical DVA test can be used for the assessment of the visual field instability referred to as disabling. PMID- 20015790 TI - Expression of the receptor activator for nuclear factor-kappaB ligand and osteoprotegerin in chronic otitis media. AB - BACKGROUND: The receptor activator for nuclear factor-kappaB ligand (RANKL) and osteoprotegerin (OPG) are the key factors controlling the osteoclast and osteoblast action in the bone. PURPOSE: The study objective was to investigate the expression level of RANKL and OPG in cholesteatoma and granulation tissue, and to assess the relationship between their expression levels and osteolysis. MATERIAL AND METHODS: Patients with chronic otitis media with cholesteatoma (n = 28) and without cholesteatoma (n = 24) treated surgically at the Department of Otolaryngology of the Medical University of Gdansk were included in the study. RANKL and OPG expressions were analyzed by immunohistochemistry and Western blot. RESULTS: RANKL and OPG were expressed in all cholesteatoma and granulation tissues. RANKL expression was mainly observed in cholesteatoma subepithelial stroma, whereas OPG-positive cells originated from the epithelium. The number of OPG-positive cells in the normal skin was significantly higher than in cholesteatoma tissues. The RANKL protein level in cholesteatoma tissues was 1.8- and 1.5-fold higher than in the auditory canal skin and granulation tissues, respectively. The number of RANKL-positive cells in cholesteatoma tissues was significantly higher than in the normal skin. No substantial differences were found in average OPG protein levels between cholesteatoma tissues and the normal auditory canal skin. The ratio of RANKL/OPG was significantly higher in cholesteatoma tissues (2.93 +/- 0.79) than in the skin samples (1.36 +/- 0.34). CONCLUSIONS: Altered ratio of RANKL/OPG protein level in cholesteatoma tissues suggests that these proteins might be somehow involved in the pathogenesis of cholesteatoma. However, to resolve this issue a study on a larger group of patients should be conducted. PMID- 20015791 TI - Shifting trends: mastoiditis from a surgical to a medical disease. AB - OBJECTIVE: The aim of this study is to review the outcome of pediatric patients suffering from acute mastoiditis treated conservatively and to correlate this to the evolution of our understanding of the shift in which mastoiditis has been transformed from a surgical to a medial disease. METHODS: We performed a retrospective review patient files hospitalized in our tertiary-care center between 2005-2008. We examined the data concerning the infection which included: presenting signs/symptoms, prior otologic history, treatment (including both surgical and conservative) prior to hospitalization and during hospitalization, computed tomography (CT), hospital duration, complications and overall outcome. This data was analyzed and compared between different patients who underwent different treatment strategies. RESULTS: Fifty-one patients were included in this retrospective review. Initially, forty-nine patients admitted to our hospital were treated conservatively. This treatment included intra-venous antibiotics, myringtomy and if needed subperiosteal abscess incision and drainage. Only 2 patients underwent CT scanning on admission. Further on, during hospitalization 4 additional patients underwent CT scanning due to continued fever or progression of local disease. All four CT scans showed no intra-cerebral complications, and so all continued with conservative treatment. CONCLUSION: Most cases of acute mastoidits may be treated with a conservative therapy regime. This regime, in our opinion, should include three branches: the first intravenous antibiotic therapy using a broad spectrum antibiotic. The second is myringotomy and the third branch is incision and drainage of subperiosteal abscess when needed. PMID- 20015792 TI - Benign paroxysmal positional vertigo after decompression sickness: a first case report and review of the literature. AB - Benign paroxysmal positional vertigo is a common cause of vertigo. We describe a previously unreported case of this clinical entity in a young, fit recreational water diver, having experienced decompression illness. Full recovery occurred after hyperbaric recompression therapy, and he remained symptom free on 6-week follow-up. We review the literature and discuss the pathogenesis of benign paroxysmal positional vertigo, proposing that semicircular canal nitrogen bubble formation could have been the primary etiological event leading to this condition. PMID- 20015793 TI - Surgical outcomes and histology findings after tonsillectomy in children with periodic fever, aphthous stomatitis, pharyngitis, and cervical adenitis syndrome. AB - PURPOSE: This study aimed to evaluate (a) specific histologic findings in children with periodic fever, aphthous stomatitis, pharyngitis, and cervical adenitis (PFAPA) syndrome who had tonsillectomy and (b) to assess any improvement of symptoms after tonsillectomy with or without adenoidectomy. MATERIALS AND METHODS: This project is a retrospective study performed at "Aghia Sophia" Children's Hospital (Athens, Greece), between May 2007 and July 2008. Nine children were recruited into this study. Patients with defined diagnostic criteria for PFAPA syndrome till 14 years of age undergoing tonsillectomy with or without adenoidectomy were included in the study. RESULTS: Nine children met our inclusion criteria in the PFAPA group, of which 5 were male (55.56%) and 4 were female (44.44%), with ages ranging between 2.5 and 5 years at the age of surgery (mean, 3.4 years). The length of follow-up ranged from 6 to 19 months. Eight (88.89%) of 9 patients had complete remission of symptoms immediately after surgery. The histologic and immunohistochemical examination showed features of chronic tonsillar inflammation. CONCLUSIONS: (a) No specific findings were observed in the tonsils of PFAPA patients; (b) we could not find any difference in tonsillar histology between PFAPA and chronic tonsillar inflammation; and (c) tonsillectomy is an effective treatment for PFAPA syndrome, improving patients and parental quality of life. PMID- 20015794 TI - Novel Epstein-Barr virus immunoglobulin G-based approach for the specific detection of nasopharyngeal carcinoma. AB - PURPOSE: This study was designed to find a reliable Epstein-Barr virus (EBV) immunoglobulin (Ig) G-based diagnostic/screening test for nasopharyngeal carcinoma (NPC) able to demarcate between the NPC-related seropositivity of EBV IgG antibodies and that of other head and neck cancer (HNCA) and control groups. The NPC-associated immunosuppression affects EBV IgA much more than IgG, leading to inconsistent detection of NPC using EBV IgA antibodies. MATERIALS AND METHODS: One hundred twenty-two HNCA patients, 42 NPC, 66 laryngeal carcinoma, and 14 hypopharyngeal carcinoma and 3 groups of 100 control subjects were enrolled in this study. Enzyme-linked immunosorbent assay (ELISA) was used to find a specific cutoff value for the NPC-related seropositivity of EBV IgG antibodies. RESULTS: NPC group showed higher serum level of EBV IgG antibodies than control and other HNCA groups (P < .05). However, the traditional cutoff value, mean + 2 SDs of control subjects, failed to demarcate the seropositives of NPC patients from those of healthy population (P > .05). The new cutoff value, mean + 2 SDs of the seropositives group of control subjects who had already been grouped by the traditional cutoff value, proved successful. It succeeded to demarcate between the NPC-related EBV IgG seropositivity and that issued from the persistent, latent, or reactivated EBV infection in the population (P < .05). The sensitivity/specificity of NPC detection by the new cutoff-based ELISA kit, 76.19% and 86%, was close or higher than that of EBV IgA antibodies. CONCLUSION: EBV IgG-based ELISA could be used for the diagnosis of NPC using a new cutoff threshold that excludes the population baseline of EBV IgG seropositivity. PMID- 20015796 TI - Spurious decline in intraoperative parathyroid hormone: false positives in parathyroid surgery. AB - OBJECTIVES: The aims of this study were to (1) describe a false-positive result using a highly sensitive intraoperative parathyroid hormone (PTH) assay in an adult patient with primary hyperparathyroidism and (2) discuss the potential pitfalls of revision parathyroid surgery and the implication of various localization techniques described in the literature. METHODS: A case report is described from a tertiary care university hospital. A literature review detailing diagnostic tools used to improve outcomes in parathyroid surgery is presented. The potential inaccuracies of intraoperative PTH assays are discussed. RESULTS: We present a 71-year-old woman with primary hyperparathyroidism who was referred to our institution for revision surgery. The patient had preoperative sestamibi imaging that localized a right inferior parathyroid lesion. Intraoperatively, a specimen consistent with parathyroid tissue was removed and sent for frozen section. The intraoperative PTH levels were noted to decrease from 154 pg/mL (preincision) to 28 pg/mL (20 minutes postexcision). The frozen section results were consistent with a lymph node. This stimulated a 4-gland exploration, which confirmed normal left superior and inferior parathyroid glands. A 1.5-cm right retroesophageal parathyroid was subsequently discovered and excised. Final intraoperative PTH levels were 20 pg/mL. CONCLUSION: Rapid PTH assays have become the mainstay of parathyroid surgery at many institutions; however, despite their accuracy, false-positive results are known to occur. We present a case of an inaccurate decline in intraoperative PTH and use this case report as a means to highlight some potential pitfalls of the test. PMID- 20015795 TI - Cochlear implantation in late-implanted adults with prelingual deafness. AB - PURPOSE: The purpose of this study was to examine the effect of cochlear implantation (CI) on prelingually deafened participants who were implanted as adults. The effect of the CI was examined with regard to the following variables: communication, family, social skills, education, and work satisfaction with one's life, loneliness, and self-esteem. MATERIALS AND METHODS: Thirty-eight adults participated. Four self-report questionnaires were used at 2 points in time: before and after CI. RESULTS: The research findings show significant differences in the reports of most variables before and after implantation. The participants felt better with regard to communication, social skills, education, and work and satisfaction with one's life after implantation in comparison to their feelings before implantation. Furthermore, they felt less lonely after implantation. However, there were no significant differences before and after implantation regarding their feelings within the family and regarding their self-esteem. CONCLUSIONS: The results demonstrated the need to evaluate the benefits resulting from the CI not only with traditional clinical measures but with additional measures as well. Furthermore, they demonstrated the benefit of the CI on the positive psychosociological implications of prelingually deafened adults. PMID- 20015797 TI - Perineural invasion detected by high-field 3.0-T magnetic resonance imaging. AB - The presence of perineural invasion (PNI) with cutaneous squamous cell carcinoma portends a poor prognosis. PNI may be detected radiographically or clinically, with motor or sensory deficits, and is confirmed by histologic evaluation. Recent interest has grown regarding the role of imaging in determining the presence of PNI and its preoperative implications. We report on a patient with cutaneous squamous cell carcinoma invading the parotid gland with clinical evidence of facial nerve weakness. On standard 1.5 Tesla (T) magnetic resonance imaging (MRI), bilateral parotid glands were symmetric; however, a second high-field 3-T MRI revealed asymmetric enhancement of the left facial nerve at the stylomastoid foramen and extending throughout the left parotid gland. PNI was later confirmed on histopathology. The presurgical determination of PNI was essential in our mapping the exact tumor location and in our surgical planning. PMID- 20015798 TI - Sarcoidosis of the submandibular gland: A systematic review. AB - INTRODUCTION: Submandibular gland sarcoidosis is rare and little is known about its clinical presentation besides the usual neck swelling. The aim of the study was to extract clinical knowledge on submandibular sarcoidosis from the literature. METHODS: A systematic review was performed using a search in Medline with the key-words "sarcoidosis," "submandibular," "submaxillary." RESULTS: Forty six articles fitting the search criteria were found, whereas 31 had to be excluded because they did not report submandibular gland sarcoidosis. Twenty cases of submandibular gland sarcoidosis were considered suitable for analysis. Almost all reported cases concerned female patients. In some cases submandibular gland's swelling is the first and only manifestation of the disease. CONCLUSION: Sarcoidosis should be considered in the differential diagnosis of all progressive and painless swellings of the submandibular gland, especially in women. Rarely, it may be the first manifestation of the disease. PMID- 20015799 TI - Clinicopathologic similarities between Mikulicz disease and Kuttner tumor. AB - PURPOSE: Recent studies have revealed that Mikulicz disease (MD) differs from Sjogren syndrome and is an immunoglobulin G(4) (IgG(4))-related systemic disease. Kuttner tumor (KT) is also reported to be an IgG(4)-related disease. In this study, we examined the clinicopathologic and serologic findings in MD (39 patients) and KT (6 patients) and attempted to discern the similarities between MD and KT. MATERIALS AND METHODS: We diagnosed 39 patients with MD and 6 patients with KT. We analyzed the clinicopathologic and serologic findings (IgG subclasses) in 39 patients with MD and 6 patients with KT. Submandibular and labial salivary gland specimens obtained from patients with MD and KT were stained with anti-IgG(4) antibodies. RESULTS: The mean IgG(4) concentration (+/ SD) was 931.1 +/- 796.2 mg/dL in patients with MD and 756.2 +/- 449.2 mg/dL in patients with KT. Abundant infiltration of IgG(4)-positive plasmacytes into the salivary glands was observed in both patients with MD and patients with KT. CONCLUSION: The serologic and histopathologic findings in MD and KT are very similar, and these 2 conditions may be IgG(4)-related systemic diseases. PMID- 20015800 TI - Cystic masses of the supraclavicular fossa: clinical features and diagnostic strategies. AB - PURPOSE: Cystic masses of the supraclavicular fossa (SCF) are uncommon. The diverse anatomical structures within the SCF create an extended differential diagnosis for any mass arising in the SCF. This study describes the presenting symptoms, radiologic findings, medical and surgical management, and posttreatment outcomes of various cystic mass presenting in the SCF. A review of the literature and diagnostic and therapeutic algorithms are also provided. METHODS: A retrospective study of an academic tertiary care head and neck cancer center was done. Seven cases of treated cystic masses of the SCF were identified. Diagnostic and therapeutic interventions are described. RESULTS: Cross-sectional imaging and needle aspiration or biopsy were obtained in all 7 cases. Aspirates accurately differentiated benign from malignant lesions in 6 cases, identified the offending pathology in 3 cases, and provided inoculum for culture-directed antibiotic therapy in 1 case. Surgical intervention was used for definitive therapy in 6 cases. All patients achieved complete resolution of signs and symptoms of the mass. CONCLUSIONS: Effective evaluation of a cystic lesion within the SCF mandates a thorough understanding of the anatomy and differential diagnosis. A well-defined algorithm allows successful management of benign and malignant lesions in the SCF. PMID- 20015801 TI - Harmonic Scalpel vs "cold knife" dissection in uvulopalatopharyngoplasty. AB - PURPOSE: The aim of this study has been to compare the Harmonic Scalpel (HS) and the "cold knife" dissection in the treatment of snoring by uvulopalatopharyngoplasty (UPPP). MATERIALS AND METHODS: The investigation included 40 adult males with snoring. Patients were assigned to two homogeneous groups (A and B) and UPPP was performed using HS or "cold knife" dissection in groups A and B, respectively. Before and 6 months after surgery, each patient was studied using the apnea-hypopnea index and a visual analogue scale of snore levels. Postoperative assessment of pain was evaluated on postoperative days 1 and 10. All the patients underwent to a perioperative evaluation concerning the duration of surgical dissection, the amount of intraoperative blood loss and days of hospitalization. RESULTS: In group A, the HS provided excellent control without side effects on the adjacent structures and postoperative complications. Harmonic Scalpel's group experienced shorter operation time, lesser blood loss, fewer days of hospitalization and lower postoperative pain. CONCLUSIONS: The use of the HS in UPPP is safe and confers some advantages over conventional methods of UPPP: its use led to diminished bleeding, shorter operation time, lesser pain, and better wound healing in the postoperative period. PMID- 20015802 TI - Age-related changes in the hyoepiglottic ligament: functional implications based on histopathologic study. AB - PURPOSE: The study aimed to identify age-related changes in the hyoepiglottic ligament associated with function of the epiglottis during swallowing and respiration. MATERIALS AND METHODS: Normal postmortem laryngeal tissue samples were obtained at autopsy from 20 individuals with no history of laryngeal disease. The subjects were divided into 2 groups: those aged 81-91 years (elderly group, n = 11) and those aged 31-48 years (non-elderly group, n = 9). Specimens were subjected to Elastica van Gieson and hematoxylin-eosin staining, and characteristics of the hyoepiglottic ligament were compared between groups. RESULTS: The hyoepiglottic ligament extended from the epiglottis to both lingual muscles and the hyoid bone (pars lingualis and pars hyoideus). The numbers of muscle fibers (P < .001), collagenous fibers (P < .01), and elastic fibers (P < .001) were significantly decreased in the elderly group in comparison to those in the non-elderly group. CONCLUSION: Age-related changes in the hyoepiglottic ligament appear to be associated with aspiration, obstructive sleep apnea syndrome, and acquired laryngomalacia in the elderly. PMID- 20015803 TI - Endoscopic evaluation of middle ear ventilation route blockage. AB - OBJECTIVES: To describe middle ear ventilation route blockage, relieved during middle ear endoscopic surgery, and to analyze its association with mastoid hypopneumatization/sclerotization. STUDY DESIGN: Prospective case series with intraoperative analyses, and with a case-control computed tomographic scan comparison. METHODS: Intraoperative findings during endoscopic middle ear surgery are described. Patients with middle ear ventilation route blockage were included in the study group (22 patients), while patients without middle ear ventilation route blockage were included in the control group (16 patients). An intra-patient and inter-group comparison of evaluated mastoid pneumatization was performed from the preoperative computed tomographic scans. RESULTS: Middle ear ventilation route blockage was classified into three types (A, B, C) according to intraoperative findings. Intrapatient and intergroup comparisons showed that the presence of blockages of middle ear ventilation trajectories is associated with a statistically significantly higher prevalence of hypopneumatization/sclerotization of the mastoid in the study group, a typical sign of middle ear dysventilation pathologies. CONCLUSIONS: Intraoperative evaluation of the middle ear anatomy during endoscopic surgery for inflammatory pathology allows us to clearly visualize the presence of anatomic blockages of the middle ear ventilation trajectories. These blockages might provoke a sectorial dysventilation of the middle ear, with consequent reduction of pneumatization of the mastoid. Further studies will be able to clarify to what extent selective dysventilation phenomena could be a principal factor in influencing middle ear pressure homeostasis. PMID- 20015804 TI - Tonsillectomy for the treatment of tonsillitis-induced immunoglobulin A nephropathy. AB - Tubular occlusion from red blood cell casts secondary to immunoglobulin A nephropathy (IgAN) is a rare, serious complication of tonsillitis that can cause acute renal failure, also referred to as acute kidney injury. IgAN is the most common primary glomerulonephritis with up to 20% of cases resulting in renal failure worldwide. Tonsillectomy is an effective treatment option for patients suffering from IgAN secondary to recurrent acute tonsillitis. Tonsillectomy alone or in combination with additional medical modalities improves renal function and can have a positive effect on long-term renal survival. PMID- 20015805 TI - A systematic algorithm for the management of lower lip asymmetry. AB - PURPOSE: An asymmetric smile, caused by loss of function of the lip depressors, can be functionally and cosmetically debilitating. Although some surgeons report excellent results with muscle transfer to the lower lip, many facial reanimation surgeons find that dynamic techniques do not consistently address the lower lip. Our objectives were to retrospectively review our outcomes after treatment of the asymmetric lower lip, and to propose a progressive, stepwise algorithm for the management of lower lip asymmetry in facial paralysis. MATERIAL/METHODS: Retrospective chart review was performed on all patients treated in a multidisciplinary facial nerve center with lower lip asymmetry over an eighteen month period. Treatment ranged from a temporary trial of lidocaine, to chemodenervation with botulinum toxin, to pedicled digastric muscle transfer, and/or resection of the nonparetic depressor labii inferioris (DLI). RESULTS: Fifty-seven patients were treated with chemodenervation with botulinum toxin, four with anterior belly of the digastric transfer, and 3 with DLI resection. All patients with DLI resection had undergone chemodenervation to the contralateral lower lip with botulinum toxin and were pleased with the appearance of their smile. CONCLUSIONS: We have found that lower lip asymmetry is optimally managed by adherence to a standardized protocol that offers patients insight into the likely outcome of chemodenervation or surgery and progresses systematically from the reversible to the irreversible. We present our algorithm for the management of the asymmetric lower lip, which reflects this graduated approach and has resulted in high patient satisfaction. PMID- 20015806 TI - Low-dose computed tomography of the paranasal sinuses: radiation doses and reliability analysis. AB - PURPOSE: The study aimed to (1) optimize the radiation doses of computed tomography (CT) of paranasal sinuses, (2) compare the radiation doses of different CT protocols with that of plain radiography, and (3) evaluate the reliability of low-dose CT in the detection of pathology and characterization of the detected pathology. MATERIALS AND METHODS: A head phantom was examined with different scan parameters to define a cutoff value to which the radiation dose can be reduced without negative impact on image quality. Kruskal-Wallis test and Wilcoxon W test were performed to compare the effective doses of the plain radiography in 30 patients with that of 3 different CT protocols in a total of 90 patients. The interobserver and intraobserver agreement in the detection of pathologic findings and in characterization of the pathology was estimated by calculating kappa value. RESULTS: The effective doses of plain radiography and low-dose CT were 0.098 and 0.045 mSv, respectively (P < .001). The effective dose of standard CT of sinuses (0.371 mSv) was 3.8 times higher than that of plain radiography and 8.2 times higher than that of low-dose CT (P < .001). The interobserver and intraobserver agreement on CT with regard to detection of pathology and pathology characterization was almost perfect (kappa values 0.81-1) compared to fair (kappa values 0.38-0.39) in plain radiography. CONCLUSIONS: The here proposed low-dose CT means significant dose reduction and is a reliable method in the investigation of the paranasal sinuses. PMID- 20015807 TI - Endoscope-assisted intra-oral resection of the external thyroglossal duct cyst. AB - OBJECTIVE: Surgical removal of a thyroglossal duct cyst (TGDC) and its tract is usually accomplished through an external neck incision, including the removal of the middle part of hyoid bone and a block of tissues extending to the foramen cecum. However, this procedure inevitably results in neck scarring. We implemented a modified approach to TGDC removal in a 28-year-old woman through the floor of the mouth using an endoscope system. CASE REPORT: Here, we describe the detailed procedure of the endoscope-assisted intra-oral resection for TGDC. The total operative time was 130 minutes. The patient complained of swelling and pain in the floor of the mouth for 2 days, but did not require any intervention. Follow-up imaging studies confirmed no recurrence (18 months) without any sequelae, and the patient was satisfied with her surgical outcome. CONCLUSION: The intra-oral approach through the floor of the mouth is a technically feasible alternative surgical option that allows for complete removal of a TGDC without the neck scar. PMID- 20015808 TI - Antioxidant micronutrient impact on hearing disorders: concept, rationale, and evidence. AB - PURPOSE: Although auditory disorders are complex conditions, device-related modalities dominate current treatment. However, dysfunction from the central cortex to the inner ear apparatus is increasingly thought to be related to biochemical pathway abnormalities and to free radical-induced oxidative damage and chronic inflammation. Therefore, considering appropriate biologic therapy as an adjunct to standard care against these damaging factors may provide rational expansion of treatment options for otolaryngologists and audiologists. METHODS: This review outlines the biologic concepts related to some auditory and vestibular conditions and details the current rationale for utilizing antioxidants for a spectrum of hearing disorders. The strategy is based on the authors' collective experience in antioxidant science and supported with published research, pilot animal data and preliminary clinical observations. RESULTS: A comprehensive micronutrient approach was developed to exploit these pathways, and demonstrated safety and efficacy against oxidative damage and inflammation and clinically relevant neuroprotection. Cooperative research with Department of Defense institutions used prospective, randomized designs to show (1) reduction in oxidative damage measured in plasma and urine over six months, (2) protection against oxidative damage during 12 weeks of intense military training, (3) protection against inflammation after total body blast exposure (rodents), (4) strong neuroprotection against chemically-induced Parkinson's disease (rodents), (5) nerve VIII function improvement after concussive head injury in military personnel, and (6) tinnitus improvement in majority of patients after 90-day evaluation. CONCLUSION: This systematic review of biologic strategies against hearing disorders combined with new animal and human observations may provide a rational basis for expanding current practice paradigms. PMID- 20015809 TI - A prolonged buried fish bone mimicking Ludwig angina. AB - Most migrated foreign bodies in the neck were removed immediately in patients with persistent symptoms. It is a rare condition that a fish bone was buried for a prolonged time in the tongue with little discomfort. We report a unique case of an ingested fish bone lodged in the tongue for 16 months until infection ensued. Ludwig angina was considered first because the patient had fever, odynophagia, swelling of the tongue, and mouth floor. The fish bone buried in the tongue was incidentally found on the computed tomography scan and successfully removed by surgical exploration. Although dental infection is the most common underlying cause in Ludwig angina, embedded foreign body should be considered as one of the pathogenesis. On the other hand, computed tomography scan can be useful in identifying extraluminal migration of fish bones in the neck. PMID- 20015810 TI - Simvastatin and Ginkgo biloba in the treatment of subacute tinnitus: a retrospective study of 94 patients. AB - OBJECTIVES: Studies suggest that hypercholesterolemia promotes the development of inner ear disorders such as tinnitus. However, the underlying pathomechanisms are still not clearly defined. METHODS: A retrospective study was performed to assess whether a reduction of serum cholesterol by 3-hydroxy-3-methylglutaryl coenzyme A reductase inhibitors may result in a relief of subacute tinnitus. Remission rates of 58 patients were investigated after 4 months of treatment with simvastatin (40 mg). Results were compared to treatment with Ginkgo biloba (120 mg; n = 36) as control group. Differences between tinnitus score at the day of first treatment and after 4 months were used as main outcome measure. RESULTS: After treatment with simvastatin or G biloba, tinnitus score decreased from 41.3 +/- 10.4 to 37.4 +/- 17.3 and from 44.7 +/- 11.2 to 41.2 +/- 8.7, respectively. However, independently of the treatment regimen, differences of tinnitus scores were considered not significant. CONCLUSIONS: After administration of simvastatin over 4 months, this retrospective study has shown no significant efficacy in treatment of subacute tinnitus. For a more conclusive answer, further prospective, double blind, and placebo-controlled studies with a larger number of patients are needed. PMID- 20015811 TI - Isolated malleus-handle fracture surgical repair using tragal cartilage. AB - The isolated malleus-handle fracture is a rare ossicular injury and tends to be overlooked when the tympanic membrane appears normal. Various surgical attempts have been made to correct this fracture; however, these techniques largely relied on xenograft implantation materials; the autologous cartilaginous graft application has never been reported. Herein we describe a simple, rapid, and effective method that uses available tragal cartilaginous graft to repair an isolated malleus-handle fracture. Our approach offers a reliable choice to restore continuity of the ossicular chain and produce a more satisfying, subjective hearing ability in this uncommon fracture situation. PMID- 20015812 TI - Comparison between topical honey and mafenide acetate in treatment of auricular burn. AB - The auricle is a frequently injured part of the head and neck during thermal injury leading to ear deformity. The burned ear represents one of the most difficult problems for reconstructive surgeons. Mafenide acetate is a topical agent used routinely for these patients, but it has some disadvantages including painful application and allergic rash. Some authors have reported the healing effect and antibacterial activity of honey. The study reported here was undertaken to compare the effect of honey and mafenide acetate on auricular burn in rabbit. In our study, although the pathologic score of the honey group was better than that of the mafenide group both on 14 and 21 days after burning, it was not statistically significant. In the mafenide acetate group, deep complication of burn (chondritis) was significantly lower than that of the honey group. In conclusion, in contrast to healing and antibiotic activity reported for honey, it may have failure in preventing deep bacterial complications of wound (like chondritis). So in deep wounds, the use of honey as dressing is not recommended. PMID- 20015814 TI - Competition and cooperation between nurse anesthetists and anesthesiologists: win win, win-loss, or loss-loss game in Taiwan? PMID- 20015813 TI - Unilesional pemphigus vulgaris of the scalp after cochlear implantation. PMID- 20015815 TI - Anesthesia does not increase the rate of bowel perforation during colonoscopy: a retrospective study. AB - OBJECTIVE: To compare the complication rates of bowel perforation during colonoscopy performed with or without anesthesia. METHODS: We retrospectively analyzed 9501 case records of colonoscopy performed at the Chi Mei Medical Center between 2000 and 2004, and compared the rates of bowel perforation between patients receiving anesthesia during the procedure versus those that did not receive anesthesia. Poisson distribution was used for statistical analysis. RESULTS: Only one case of perforation was found in the non-anesthetized group (n = 2460) compared with two cases in the anesthetized group (n = 7041). No statistical difference was found (p = 0.6173; risk ratio = 0.6988; 95% confidence interval by Byar's method = 0.063-7.705). CONCLUSION: During colonoscopy, anesthesia per se does not affect the rate of bowel perforation. PMID- 20015816 TI - Optimizing epidural fentanyl loading dose for early labor pain. AB - OBJECTIVE: Sufficient sensory blockade between L1 and T10 is required to relieve visceral pain during early labor. We examined whether the addition of fentanyl to a loading dose of 0.0625% bupivacaine could provide dose-dependent analgesic effects on early-stage labor pain. METHODS: Sixty parturients who requested epidural analgesia for labor pain were enrolled and randomly allocated to one of three groups. Group A (n = 20) received 10 mL of 0.0625% epidural bupivacaine as a loading dose alone. Group B (n = 20) received the same bupivacaine loading dose in combination with 2 microg/mL fentanyl. Group C (n = 20) received the same loading bupivacaine dose plus 4 microg/mL fentanyl. All patients received diluted bupivacaine plus 2 microg/mL fentanyl at a rate of 10 mL/hr as a maintenance dose. Fifteen minutes later, we recorded the highest cephalic and lowest caudal anesthetized dermatomes, side effects, and the number of patients who asked for supplemental analgesia. RESULTS: The highest anesthetized cephalic dermatome was at the level of T12 (T9-L1) in Group A, T9 (T8-T12) in Group B and T7 (T5-T9) in Group C (p < 0.05 among the three groups). Eleven patients (55%) requested supplemental bupivacaine for inadequate analgesia in Group A, six in Group B (30%), and none in Group C (0%). Pruritus was reported by seven (35%) patients in Group B and eight (40%) patients in Group C, but none in Group A. CONCLUSION: The addition of fentanyl to epidural bupivacaine dose-dependently increased the analgesic effect and higher loading doses of fentanyl increased the dermatomic coverage. We suggest that 0.0625% bupivacaine plus 4 microg/mL fentanyl is the ideal loading dose to provide the greatest segmental analgesia for early labor pain with minimal side effects. PMID- 20015817 TI - Optimal duration of urinary catheterization after thoracotomy in patients under postoperative patient-controlled epidural analgesia. AB - BACKGROUND: Epidural analgesia is widely used for efficient pain relief after major surgery. However, it may cause urinary retention, leading to delayed removal of bladder catheters with prolonged patient discomfort. Using a specific regimen in patient controlled epidural analgesia (PCEA), we examined the optimal duration of urinary catheterization in patients undergoing major thoracic surgery. METHODS: Seventy-eight patients scheduled for elective thoracotomy were prospectively randomized into two groups: Group 1, removal of the transurethral catheter on the first postoperative day (n = 38); Group 2, removal of the catheter after discontinuation of PCEA (n = 40). The PCEA regimen was a mixture containing low-dose morphine, bupivacaine and neostigmine and was given for 3 days after surgery in all subjects. Micturition problems, pain scores assessed by the visual analog scale (VAS), and side effects were evaluated during and after PCEA treatment. RESULTS: The average duration of urinary drainage after surgery was 30.2 + or - 5.1 hours and 78.5 + or - 7.3 hours in Groups 1 and 2, respectively. After removal of the bladder catheter, no patient in either group required re-catheterization for urinary retention or encountered catheter-related infection. VAS scores were significantly lower in Group 1 at rest and at 24, 36 and 48 hours after cessation of PCEA. VAS scores were significantly higher in Group 2 patients, possibly due to catheter-induced pain related to prolonged catheterization. CONCLUSION: Routine continuous bladder catheterization may not necessarily be required after thoracotomy in patients with ongoing continuous thoracic epidural analgesia. PMID- 20015818 TI - Patient satisfaction with anesthesia care in Taiwan: development and validation of a pilot version of a psychometric questionnaire. AB - BACKGROUND: To date, no pilot questionnaire of perioperative anesthetic care has been developed based on psychometric methodology in Taiwan. We describe the development and qualitative and quantitative validation of a pilot version of a psychometric questionnaire designed to measure patient satisfaction with perioperative anesthetic care in Taiwan. METHODS: A rigorous protocol was followed and involved expert consultation, literature review, development of the interview guide, semi-structured in-depth interviews, pretest and application of Aiken's two quantitative methods to determine the content validity coefficient (V value) and homogeneity reliability coefficient (H value) of each item, and the questionnaire as a whole, to ensure the pilot questionnaire showed high-content validity and reliable homogeneity. RESULTS: Our final pilot questionnaire contained six dimensions with 32 items; five of the domains were similar to those reported in the literature. One dimension entitled anesthesia-related sequelae was new. The V values for the 32 items ranged from 0.80 to 0.97 (p < 0.01) and the mean value (V) of the questionnaire as a whole was 0.90 (p < 0.01). The H values ranged from 0.55 to 0.88 (p < 0.01) and the mean H value (H) of the questionnaire as a whole was 0.71 (p < 0.01). CONCLUSION: We have developed a valid and reliable pilot questionnaire to measure patient satisfaction with perioperative anesthetic care in Taiwan. The final version of the pilot questionnaire is a 32-item instrument with six dimensions, one of which, anesthesia-related sequelae, is new. PMID- 20015819 TI - Analysis of manpower and career characteristics of nurse anesthetists in Taiwan: results of a cross-sectional survey of 113 institutes. AB - BACKGROUND: Manpower and the quality of nurse anesthetists (NA) have become critical concerns in Taiwan over the past few decades because of increasing clinical demand and the supervision of NAs by anesthesiologists. To understand manpower distribution, clinical load, job description and limitations, and job satisfaction of NAs, we conducted a cross-sectional survey in Taiwan in 2005. METHODS: The structure of the questionnaire was initially developed by a drafting group that included members of the Taiwan Society of Anesthesiologists and the Taiwan Association of Nurse Anesthetists. The validity and reliability of the questionnaire was evaluated by specialists. The survey contained questions regarding the demographic characteristics of manpower (anesthesiologist/NA ratio), clinical work load, present job roles, professional expectations, job satisfaction, and reasons for career transfer. The questionnaires were mailed to the superintendents or matrons of NAs, and the administrators of anesthesiology departments across 228 institutions with different accreditation levels and 1953 NA staff between February 1 and December 31, 2005. RESULTS: The validity and reliability of the questionnaire for the department chief and anesthesiology nursing staff was 0.8 and 0.7, respectively. Questionnaires were returned by 113 executives (39 anesthesiology department directors, 74 NA superintendents or matrons) with a response rate of 49.6%, and from 1452 NAs with a response rate of 74.3%. The average clinical load (2002-2004) for the anesthesiologists was 1500 1700 cases/year and 350-380 cases/year for the NAs. The manpower ratio of attending anesthesiologists to NAs was 1:4.3, while the medical centers held the highest ratio. The job stipulation for NAs in Taiwan was compatible with that in the United States and there was a high consistency of opinions between the directors and NA superintendents or matrons. The average rate of career transfer was relatively low (5.5%). From the executives' view, the concerns regarding management of NAs included limited staff capacity, recruiting difficulty, shortage of staff and undefined roles for NAs. On the other hand, NAs showed relatively high job satisfaction and work acceptance and a lower turnover rate in comparison with the general nursing staff. CONCLUSION: This study represents the first large-scale assessment of the distribution, clinical load, and job satisfaction for NAs in Taiwan. The roles of NAs, which include preoperative preliminary preparation and postoperative intensive care, need to be more well defined. To improve the quality of NAs and anesthetic care in Taiwan, it is vital to establish an official accreditation system and formal education programs, to institute well-defined and standardized job descriptions, and to improve resource allocation for NAs. PMID- 20015820 TI - Iatrogenic left internal iliac artery perforation during lumbar discectomy. AB - Iatrogenic intra-abdominal vascular injury can result from lumbar discectomy via the posterior approach. Although it is well known and documented in the literature, few anesthesiologists have personal experience with this life threatening incident. Here, we report a patient who sustained perforation of the left internal iliac artery at the L(4-5) level during posterior lumbar discectomy. The patient experienced refractory hypotension with tachycardia at the end of surgery, even with prompt fluid resuscitation and medical treatment. Abdominal distension and tenderness of the left lower abdominal quadrant were also noted. Emergency laparotomy was performed by the consulting vascular surgeon and revealed perforation of the left internal iliac artery. The vascular injury was successfully repaired. It is important that, as anesthesiologists, we must be aware of this potentially fatal complication. Prompt diagnosis and immediate laparotomy to control hemorrhage can result in favorable outcomes. PMID- 20015821 TI - Temporomandibular joint dislocation during orotracheal extubation. AB - Temporomandibular joint (TMJ) dislocation can occur whenever the mouth is opened wide during upper airway manipulation, even without external force. In the perioperative period, the majority of TMJ dislocations occur during anesthetic induction. We report the occurrence and management of bilateral TMJ dislocation upon orotracheal extubation in a 35-year-old woman. At the end of an otherwise unremarkable uterine myomectomy under general anesthesia, with the patient having regained consciousness in the operating room, we asked the patient to open her mouth so we could extubate her. Immediately after orotracheal extubation, however, the patient was found to be unable to close her mouth. The diagnosis of bilateral TMJ dislocation was made. The bilateral TMJ dislocation was reduced with the traditional transoral approach under morphine analgesia/sedation. The dislocation possibly occurred in consequence of her compliance with our request to open her mouth excessively before she was fully awake. As TMJ dislocation is a possible complication of upper airway manipulation, anesthesiologists should be prepared for its occurrence and to manage it competently. PMID- 20015822 TI - Eisenmenger's syndrome in pregnancy: use of Proseal laryngeal mask airway (PLMA) and epidural analgesia for elective cesarean section. AB - We describe the successful anesthetic management of cesarean section in a patient with Eisenmenger's syndrome secondary to an atrial septal defect. Although conception is discouraged in women with Eisenmenger's syndrome, in inevitable circumstances, careful and meticulous planning of anesthesia can help the parturient survive the ordeal of a cesarean section. The cardiac output must be maintained and systemic vascular resistance must not be allowed to fall. This should ensure that there is minimal change in the right to left shunt. In our patient, the scenario of Eisenmenger's syndrome was complicated by biventricular hypertrophy. We achieved the goals in our patient by using general anesthesia with the Proseal laryngeal mask airway, a combination of ketamine and propofol, and epidural analgesia. PMID- 20015823 TI - General anesthesia in a juvenile with attention-deficit hyperactivity disorder accompanied by long-term use of methylphenidate (Concerta). AB - Methylphenidate, a central stimulant, is used in the treatment of individuals who have attention-deficit hyperactivity disorder (ADHD). ADHD is a notorious worldwide disorder with a prevalence rate of 8-12% in schoolchildren, which is characterized by hyperactivity, impulsivity, and inattention. Currently, there have been few reports in the anesthetic literature examining ADHD patients who have had long-term use of methylphenidate, especially the extended-release formulation. Here, we report a case of a 14-year-old boy with ADHD treated chronically with the long-acting form of methylphenidate (Concerta), and who was scheduled to receive orthopedic surgery under general anesthesia. No significant problems or fluctuations in hemodynamics were encountered during anesthesia induction, maintenance, and emergence. The patient made an uncomplicated recovery and was discharged 3 days later without incident. PMID- 20015824 TI - Recurrent attacks of post-intubation right upper lobe atelectasis. AB - A 22-year-old woman with no history of asthma developed an acute recurrent attack of severe bronchoconstriction and right upper lobe atelectasis immediately after laryngoscopy and endotracheal intubation. The first attack had taken place 2 months earlier under identical circumstances. Induction of anesthesia for tracheal intubation was achieved using propofol, fentanyl, and rocuronium. Bronchial obstruction and bronchial intubation were excluded by bronchoscopy. The atelectasis was quickly resolved with mechanical ventilation and spasmolytic treatment on both occasions. The surgical procedure could proceed soon after resolution of the atelectasis. PMID- 20015825 TI - Critical debate on establishing a scheme for nurse anesthetists in Japan. PMID- 20015826 TI - Distribution of oxytocin-immunoreactive neuronal elements in the rat spinal cord. AB - We investigated the distribution of oxytocin in rat spinal cord using immunocytochemistry and radioimmunoassay (RIA). Each segment of the spinal cord from cervical to coccygeal contained oxytocin-immunoreactive fibers. The Rexed laminae I and II of the dorsal horn showed moderate to intense immunoreactivity. A dense network was found around the central canal where some fibers apposed the ependyma. The autonomic centers of the spinal cord at the thoracolumbar and sacral segments were heavily innervated. Few fibers were found around the motoneurons. In the white matter, the immunoreactivity was localized mainly in the dorsal part of the lateral funiculus, in the pars funicularis of the nucleus intermediolateralis and in a longitudinal network of the lateral funiculus below the spinal cord surface. Some fibers from this network entered the pia mater. RIA measurements revealed that the cervical spinal cord had lower oxytocin content than that found in either the thoracic, lumbar, sacral or coccygeal region. Our results show that the distribution of oxytocin-immunoreactive fibers in the spinal cord correlates with anatomic locations related to nociceptive, autonomic and motor functions. We assume that oxytocin-containing axons play a role in secreting oxytocin directly into the liquor space of the spinal cord. PMID- 20015827 TI - Spatial pattern analysis of nitrergic neurons in the myenteric plexus of the duodenum of different mammalian species. AB - Nitrergic myenteric neurons are especially susceptible to the development of neuropathy in functional gastrointestinal disorders. Investigations of the similarities and dissimilarities in the organization of nitrergic neurons in the various mammalian species are therefore important in an effort to determine the extent to which the results obtained in different animal models can be generalized. In the present work, the density and the spatial organization of the nitrergic neurons in the myenteric plexus of the duodenum were investigated in 7 mammalian species. After nicotinamide adenine dinucleotide phosphate diaphorase (NADPH-d) histochemistry, the Plexus Pattern Analysis software (PPAs) was applied to count the nuclei of nitrergic neurons, calculate the proportions of the areas covered by the plexus and perform randomization analysis. All 7 species exhibited a large population of nitrergic myenteric neurons, with densities in the range 12 56 cells/mm 2 . The distribution patterns of these neurons differed markedly in the different species, however, the rat was the only species in which the nitrergic neurons appeared to be randomly distributed. The PPAs in conjunction with NADPH-d histochemistry proved to be a simple and fast tool with which to reveal similarities and dissimilarities in the spatial arrangement of the nitrergic neurons in the different species. PMID- 20015828 TI - Altered open field behavior in rats induced by acute administration of 3 nitropropionic acid: possible glutamatergic and dopaminergic involvement. AB - 3-nitropropionic acid (3-NP), a substance used for modelling Huntington's disease, was given to male Wistar rats in a single 20 mg/kg b.w. dose, and the resulting behavioral alterations in spontaneous locomotor activity were measured after 30 minutes. To detect the involvement of neurotransmitter systems in this immediate effect, the NMDA antagonist MK-801 (0.8 mg/kg); as well as an agonist, quinpirole (QP, 5 mg/kg) and an antagonist, sulpiride (SP, 80 mg/kg) of the dopamine D2 receptors, were given before 3-NP to separate groups of rats. Controls were given saline. All substances were injected ip. 3-NP decreased the rats' locomotor, especially vertical, activity, whereas local activity was increased. Based on the further changes of 3-NP effects in the combination groups it could be concluded that dopaminergic rather than glutamatergic mechanisms were possibly involved in the acute behavioral effect of 3-NP. PMID- 20015829 TI - Influence of up-regulated renin-angiotensin system on the exploration, anxiety related behavior and object recognition. AB - The renin-angiotensin system (RAS) plays an important role in the development of hypertension and has serious consequences on behaviour. The aim of our study was to investigate the effect of hypertension, induced by up-regulated RAS, on the exploration, anxiety-related behaviour and object recognition in laboratory rats. In the experiment, 12 weeks old normotensive Sprague-Dawley (SD) and hypertensive TGR(mREN2)27 (TGR) male rats with up-regulated RAS were used. In the open-field test, the TGR rats were less active in ambulating, rearing and sniffing and more active in self-grooming and urinating than SD ones. In the elevated plus-maze test, the TGR rats showed lower frequency of total arm entries, closed arm entries and higher frequency of defecation than in controls. In the emergence test, TGR rats did not show significant differences. In the novel object recognition task, the TGR rats spent less time with exploration of both familiar and unfamiliar objects but preferred the novel object over the familiar one and exhibited higher percentage of the total exploring time spent with novel object exploration than SD rats. Our results indicate that the TGR rats are less actively exploring, show some modifications of emotional/anxiety-related behavior and exhibited better recognition abilities. PMID- 20015830 TI - Disruption of mitochondrial membrane potential by ferulenol and restoration by propolis extract: antiapoptotic role of propolis. AB - This paper reports an investigation of the ability of propolis extract (a resinous substance collected by honeybees from various plant sources) to restore the collapse of mitochondrial membrane potential induced by ferulenol, a sesquiterpene prenylated coumarin derivative isolated from the plant Ferula vesceritensis . We show that ferulenol was able to induce the permeability transition pore (PTP) opening. This effect is caused by the interaction of the compound with the mitochondrial respiratory chain, more particularly by the fall of membrane potential and the inhibition of complex II. We have previously demonstrated that this inhibition results from a limitation of electron transfers involved in the respiratory chain and initiated by the reduction of ubiquinone. We hypothesized that the protective effect of propolis could be due to a direct action on mitochondrial functions. So we have investigated in vitro the mitochondrial effects of Algerian propolis using rat liver mitochondria, by analysing their effects on membrane potential, mitochondrial respiration and mitochondrial swelling. We show that propolis extract was able to restore the fall of mitochondrial membrane potential. Taken together these data reveal that propolis extract may be an interesting inhibitor of PTP and provide an additional mechanism by which the natural product propolis extract may restore the mitochondrial membrane potential and to prevent apoptotic process. PMID- 20015831 TI - Effects of melatonin administration on plasma leptin concentration and adipose tissue leptin secretion in mice. AB - Both melatonin and leptin show a circadian variation in circulating levels and participate in energy metabolism. An interrelationship between these two hormones has thus been proposed. In addition, melatonin has been shown to be capable of influencing circulating leptin concentration. However, whether melatonin will increase or decrease leptin production is still uncertain. This study was undertaken to examine the effect of melatonin on leptin production using male C57BL/6 adult mice treated with or without daily melatonin supplements (10 mug/mL) in drinking water for 1 month. In addition, in vitro experiments using adipose tissue fragments derived from epididymal fat pads of adult mice incubated with or without melatonin (1 nM) administration were also conducted. The results showed that melatonin-supplemented mice had significantly higher plasma leptin levels than control mice. However, melatonin incubation did not cause any marked changes in the amount of leptin secreted from adipose tissue fragments. Our findings from this study indicate that melatonin does not affect leptin secretion via mouse adipose tissue. Nevertheless, melatonin could still influence leptinemia indirectly via regulatory effects in intact animals. PMID- 20015832 TI - Toxic nuclear effects of the organophosphorus insecticide Dichlorvos (DDVP) in human peripheral blood lymphocytes. AB - Organophosphorus insecticides are used worldwide in the control of agricultural, household and veterinary pests. Dichlorvos (2,2-dichlorovinyl dimethyl phosphate) is a commonly used organophosphorus insecticide. In this study, in order to elucidate the toxic nuclear effects of dichlorvos, freshly isolated human peripheral blood lymphocytes were incubated with 5, 10, 20, 40, 80 and 100 microg/mL of dichlorvos. According to the results, dichlorvos induced micronuclei, decreased the mitotic and replication indexes. It is a genotoxic product causing chromosomal damage (an increase in micronucleus) and cell death (decrease in mitotic and replication indexes). PMID- 20015833 TI - Photosystem II functionality and antioxidant system changes during leaf rolling in post-stress emerging Ctenanthe setosa exposed to drought. AB - We studied the changes in antioxidant system and chlorophyll fluorescence parameters in post-stress emerging Ctenanthe setosa (Rosc.) Eichler (Marantaceae) plants (PSE plants) having reduced leaf area under drought stress causing leaf rolling and re-watering. PSE plants were compared to primary stressed plants (PS) in previous studies. The parameters were measured at different visual leaf rolling scores from 1 to 4 (1 is unrolled, 4 is tightly rolled and the others is intermediate form). Water potentials and stomatal conductance of leaves were gradually decreased during leaf rolling. Similarly, maximum quantum efficiency of open PS II center and quantum yield of PS II decreased during the rolling period. Non-photochemical quenching of chlorophyll fluorescence decreased at score 2 then increased while photochemical quenching did not change during leaf rolling. Electron transport rate decreased only at score 4 but approximately reached to score 1 level after re-watering. Superoxide dismutase activity was not constant at all leaf rolling scores. Ascorbate peroxidase, catalase and glutathione reductase activities generally tended to increase during leaf rolling. Lipid peroxidation and H 2 O 2 content increased at score 2 but decreased at the later scores. On the other hand, O 2 .- production increased during the rolling period. After re-watering of the plants having score 4 of leaf rolling, antioxidant enzyme activities were lower than those of score 1. Other physiological parameters also tended to reach the value of score 1. The results indicated that PSE plants gained drought tolerance by reducing leaf area effectively induced their antioxidant systems and protected the photosynthesis under drought stress similar to PS plants. PMID- 20015834 TI - Interactive effects of salinity, nitrate, light, and seed weight on the germination of the halophyte Crithmum maritimum. AB - Interaction of salinity, nitrate, light, and seed weight on the germination of Crithmum maritimum was investigated. Seeds of three size categories were germinated at 0-200 mM NaCl with either 0, 5 or 20 mM KNO 3 . Experiments were done under darkness, white light, or red light. Regardless of seed weight, germination was maximal in distilled water. Under salinity, the smallest seeds showed the highest germination percentage. Salt impact was amplified by darkness, but was mitigated by nitrate supply, red light and their combination. At the same PPFD, germination of T2 seeds was higher, when exposed to red light than under white light, suggesting that germination was more influenced by the light type than by the PPFD. As a whole, not only salinity, nutrient availability, seed weight, and light, but also their interaction may control the germination of this halophyte. PMID- 20015835 TI - Improved alliin yield in somatic embryos of Allium sativum L. (cv. Yamuna safed) as analyzed by HPTLC. AB - Direct somatic embryo formation (without intervening callus) from garlic clove basal tissue was induced in which the influence of plant growth regulators (PGRs) on various explants was examined. Medium added with 2.0 mg/l 6-benzylaminopurine (BAP) and 0.5 mg/l 2,4-dichlorophenoxyacetic acid (2,4-D) were the most effective PGR combination for somatic embryo induction. It induced embryos directly in 85.5% of the basal clove explant. Callus induction was also obtained from other parts of explant and 2.0 mg/l 2,4-D induced callusing in 86.5% of the inoculated explants. Protein, amino acid and alliin content were measured in callus and in embryos. Somatic embryos had more soluble protein and free amino acid compared to callus. HPTLC analysis revealed that alliin was significantly high in somatic embryos compared to undifferentiated callus tissue; the content was even more in older embryos. The present study of Allium indicates that the event of morphogenetic development including in vitro embryogeny can effectively be analysed by monitoring the changes of biochemical profiles. PMID- 20015836 TI - Political challenges in randomized controlled trials of heroin-assisted treatment for severe heroin addiction: a Spanish experience. PMID- 20015838 TI - Glucocorticoid receptor over-expression promotes human small cell lung cancer apoptosis in vivo and thereby slows tumor growth. AB - Small cell lung cancer (SCLC) is an aggressive tumor, associated with ectopic ACTH syndrome. We have shown that SCLC cells are glucocorticoid receptor (GR) deficient, and that restoration of GR expression confers glucocorticoid sensitivity and induces apoptosis in vitro. To determine the effects of GR expression in vivo, we characterized a mouse SCLC xenograft model that secretes ACTH precursor peptides, and so drives high circulating corticosterone concentrations (analogous to the ectopic ACTH syndrome). Infection of SCLC xenografts with GR-expressing adenovirus significantly slowed tumor growth compared with control virus infection. Time to fourfold initial tumor volume increased from a median of 9 days to 16 days (P=0.05; n=7 per group). Post-mortem analysis of GR-expressing tumors revealed a threefold increase in apoptotic (TUNEL positive) cells (P<0.01). Infection with the GR-expressing adenovirus caused a significant reduction in Bcl-2 and Bcl-xL transcripts. Furthermore, in both the GR-expressing adenovirus-infected cells and tumors, a significant number of uninfected cells underwent apoptosis, supporting a bystander cell killing effect. Therefore, GR expression is pro-apoptotic for human SCLCs in vivo, as well as in vitro, suggesting that loss of GR confers a survival advantage to SCLCs. PMID- 20015839 TI - Apathy and depression in Parkinson disease. AB - The purpose of this study was to investigate the prevalence and clinical correlates of apathy and depression in Parkinson disease (PD), and to clarify whether apathy can be dissociated from depression. One hundred fifty patients with PD completed the Beck Depression Inventory Second Edition (BDI-II), Starkstein's Apathy Scale (AS), and a quality of life (QOL) battery. Hoehn and Yahr (HY) staging, the Unified Parkinson's Disease Rating Scale (UPDRS), and the Mini-Mental State Examination (MMSE) were performed on the same day. Apathy (AS score > or = 16) was diagnosed in 60% of patients and depression (BDI-II score > or = 14) in 56%. Apathy coexisted with depression in 43% of patients, compared with depression without apathy in 13% and apathy without depression in 17%. Apathy scale score was significantly correlated with UPDRS scores, HY stage, and age, whereas BDI-II score was correlated only with UPDRS scores. Both AS and BDI II scores were negatively correlated with QOL. However, multiple regression analysis revealed that depression was strongly and negatively associated with emotional well-being and communication, whereas apathy was mainly associated with cognition and stigma. These findings suggest that apathy and depression may be separable in PD, although both are common in patients with PD and are associated with QOL. PMID- 20015840 TI - Reproductive-aged women's knowledge and attitudes regarding infant-feeding practices: an experimental evaluation. AB - Using an experimental design, this study assessed knowledge, attitudes, and intentions regarding infant feeding practices among Canadian female university undergraduates (N = 285). Participants completed a survey of knowledge, attitudes, and intentions with respect to infant-feeding practices. Two versions of the survey were randomly distributed to participants: one containing a photograph of a woman breastfeeding her infant (n = 131) and the other containing a photograph of the same women bottle-feeding her infant (n = 154). Findings indicated that the majority of the sample had been breastfed (84%) and intend to breastfeed their own offspring (97%). The intention to breastfeed future offspring was predicted by knowledge, attitudes, and perceptions of social norms. Participants reported more positive attitudes toward visual depictions of breastfeeding compared with bottle-feeding but less positive views of breastfeeding in public compared with bottle-feeding in public. Participants also significantly underestimated the health benefits and optimal duration of breastfeeding. The findings highlight gaps in knowledge that may contribute to premature cessation of breastfeeding among Canadian women and suggest the need for breastfeeding education. PMID- 20015841 TI - The relationship between breastfeeding test weights and postpartum breastfeeding rates. AB - This secondary analysis was conducted to determine the relationship between test weights and days of breastfeeding. Test weights were performed with a scale (accurate to 2 grams) before and after feeding. Days of breastfeeding was measured by self-reporting. All test weights were interrelated and were associated with higher days of breastfeeding. Test weights at day 2 to 4 and at 2 weeks were not correlated significantly with either intention or self-efficacy at any time. However, 6-week test weights were correlated with intention at all 3 times, and self-efficacy at 6 weeks. Women with higher breastfeeding self efficacy scores tended to report higher intention to breastfeed for 6 months. PMID- 20015842 TI - Akt downregulation by flavin oxidase-induced ROS generation mediates dose dependent endothelial cell damage elicited by natural antioxidants. AB - High intake of natural antioxidants (NA) from plant-derived foods and beverages is thought to provide cardiovascular benefits. The endothelium plays a pivotal role in cardiovascular homeostasis, and for this reason, the molecular events resulting from NA actions on endothelial cells (ECs) are actively investigated. Here, we show the direct impact of two NA, coumaric acid and resveratrol, on intracellular reactive oxygen species levels, protein carbonylation, and cell physiology in human ECs. While at lower doses, both NA promoted antioxidant effects, at moderately high doses, NA elicited a dose-dependent pro-oxidant effect, which was followed by apoptosis, cell damage, and phospho-Akt downregulation. NA-induced pro-oxidant effects were counteracted by N-acetyl cysteine and diphenyleneiodonium (DPI), suggesting a role for flavin oxidases in NA-induced toxicity. DPI also prevented NA-induced phospho-Akt downregulation indicating that Akt can work downstream of flavin oxidases in mediating cellular responses to NA. Stimulation of phospho-Akt by insulin dramatically counteracted NA-induced cell death, an effect abolished by Akt inhibition further suggesting that mechanistically Akt regulates cell survival in response to NA-induced stress. Although further studies are required to better characterize the molecular mechanism of NA-induced cell toxicity, our study is the first to show in a human vascular model that moderately high doses of NA can induce cell damage mediated by flavoproteins and the Akt pathway. PMID- 20015843 TI - Effects of 10 cigarette smoke condensates on primary human airway epithelial cells by comparative gene and cytokine expression studies. AB - Cigarettes vary in tobacco blend, filter ventilation, additives, and other physical and chemical properties, but little is known regarding potential differences in toxicity to a smoker's airway epithelia. We compared changes in gene expression and cytokine production in primary normal human bronchial epithelial cells following treatment for 18 h with cigarette smoke condensates (CSCs) prepared from five commercial and four research cigarettes, at doses of approximately 4 microg/ml nicotine. Nine of the CSCs were produced under a standard International Organization for Standardization smoking machine regimen and one was produced by a more intense smoking machine regimen. Isolated messenger RNA (mRNA) was analyzed by microarray hybridization, and media was analyzed for secreted cytokines and chemokines. Twenty-one genes were differentially expressed by at least 9 of the 10 CSCs by more than twofold, including genes encoding detoxifying and antioxidant proteins. Cytochrome P450, family 1, subfamily A, polypeptide 1 (CYP1A1) and NAD(P)H dehydrogenase, quinone 1 (NQO-1) were selected for validation with quantitative real-time PCR (qRT-PCR) and Western blot analyses. NQO-1 expression determined with microarrays, qRT-PCR, and Western blotting differed among the CSC types, with good correlation among the different assays. CYP1A1 mRNA levels varied substantially, but there was little correlation with the protein levels. For each CSC, the three most induced and three most repressed genes were identified. These genes may be useful as markers of exposure to that particular cigarette type. Furthermore, differences in interleukin-8 secretion were observed. These studies lay the foundation for future investigations to analyze differences in the responses of in vivo systems to tobacco products marketed with claims of reduced exposure or reduced harm. PMID- 20015848 TI - Effect of cardiac resynchronization therapy on regional left ventricular function: a speckle tracking strain analysis. AB - AIMS: The mechanisms associated with cardiac resynchronization therapy (CRT) benefits have been elucidated in part, however little is known about the effects of biventricular pacing (BVP) on regional myocardial contractility. METHODS AND RESULTS: Twenty patients with conventional CRT criteria were studied before and after device implantation. Speckle tracking strain analysis was performed from standard bidimensional acquisitions during different pacing configurations. Longitudinal and transverse strains were measured for the six basal left ventricular (LV) segments. Acute CRT improved LV end-diastolic and end-systolic volumes without significantly modifying LV ejection fraction. Moreover, CRT produced significant changes in regional strain. When compared with spontaneous rhythm, simultaneous BVP caused a significant increase in longitudinal septal strain (-4.8 vs. -7.3%, P = 0.003) and a significant decrease in longitudinal lateral strain (-8.2 vs. -5.9%, P = 0.03). Simultaneous BVP also induced a significant decrease in transverse lateral strain from 17.9 to 10.9% (P = 0.004). CONCLUSION: In this acute echocardiographic study, BVP decreased lateral wall deformation while improving septal wall contraction, thus potentially improving LV contractile pattern. PMID- 20015849 TI - Incidence of left ventricular hypertrophy in children with kidney disease: impact of method of indexation of left ventricular mass. AB - AIMS: Patients with chronic kidney disease are at high risk of cardiovascular morbidity and mortality. Increased left ventricular mass (LVM) has been shown to be an adverse prognostic factor. LVM may be indexed for body size by different methods related to height, weight, or body surface area (BSA). Our Null hypothesis was that different methods of indexation would not influence categorization as to whether LVM was within normal limits or increased. METHODS AND RESULTS: Prospective study in children with renal disease. M-mode echocardiography assessed by single investigator blinded to medical therapy. Three different partition values and two different published z-scores were used: Method 1: indexation of LVM to the allometric height in metres raised to the power of 2.7 (m(2.7)), value of 38.6 g/m(2.7) denotes 95th percentile. Method 2: indexation of LVM by body weight, ratio of LVM in gm/kg of >3.0 indicating LVH. Method 3: indexation by BSA, 88.9 g/m(2) represents 95th percentile. Z-score based methods from recent studies with z-score >1.65 denoting LVH, Method 4: z score indexed either for BSA or Method 5, z-score indexed for height(2.7). One hundred and twenty-three echocardiograms were performed in 80 patients with a mean +/- SD age of 13.1 +/- 3.1 years, height 147.2 +/- 15.3 cm, weight 46.4 +/- 15.8 kg, and body mass index 20.8 +/- 4.1 kg/m(2). Method 1, LVH was observed in 68 (55.3%) studies. Method 2, 32 (26%) studies had LVM/kg >3.0. Method 3, 51 studies (41.5%) demonstrated LVH. Method 4, 22 (17.9%) studies demonstrated LVH and Method 5, 25 studies (20%) demonstrated LVH (chi(2) test, P < 0.001). CONCLUSION: Different methods of indexation have a profound influence on the categorization of children with respect to LVH. This will have a major impact on the number of patients who are treated as per current guidelines especially in high-risk groups. PMID- 20015850 TI - Disparities in emptying velocity within the left atrial appendage. AB - AIMS: Pulsed Doppler measurement of left atrial appendage (LAA) emptying velocity, a marker of left atrium contractile function, has been shown to predict success of cardioversion, thrombo-embolic risk, and maintenance of sinus rhythm after cardioversion and pulmonary vein isolation. However, in the published literature, emptying velocity measurement location is not uniform, and no standard currently exists. We assessed the hypothesis that emptying velocity when acquired near the LAA orifice differs from that at the LAA apex. METHODS AND RESULTS: The study group comprised 44 patients (32 in sinus rhythm and 12 in atrial fibrillation) who were able to complete a non-emergent transoesophageal echocardiography. Pulsed Doppler recordings were obtained with the sample volume first positioned 1 cm from the LAA orifice, and then positioned within 1 cm of the LAA apex. At each location, we calculated the average of the peak end diastolic LAA emptying velocity from five consecutive cardiac cycles. LAA orifice emptying velocity was higher than the apex emptying velocity in all patients. The median velocity at the orifice was 72 cm/s, which was 45% higher than the median velocity at the apex (43 cm/s, P < 0.001). Lower LAA emptying velocity at the orifice was associated with a larger discrepancy between orifice and apex velocities. The ratio of orifice to apex velocity did not vary with orifice velocity. Multivariate analysis demonstrated that clinical patient characteristics were not significant predictors of the discrepancy between orifice and apex velocities. CONCLUSION: LAA emptying velocity is greater at the LAA orifice compared with the LAA apex. Higher, more easily measured velocity and greater variability observed with orifice measurements make it the location of choice for research and clinical applications. PMID- 20015851 TI - Coronary sinus dilatation in an elderly patient with dyspnoea. PMID- 20015852 TI - Motor neuron disease: systematic reviews of treatment for ALS and SMA. AB - INTRODUCTION: There is no curative treatment for the common motor neuron diseases, amyotrophic lateral sclerosis (ALS) and spinal muscular atrophy. Nevertheless, there is an increasing volume of published studies. This review assesses the current evidence for treatment of these conditions. SOURCES OF DATA: Primarily, the systematic reviews of the Cochrane Collaboration, with additional reference to other systematic reviews and online sites. AREAS OF AGREEMENT: Riluzole remains the only medication with demonstrated efficacy and regulatory approval for the treatment of ALS. AREAS OF CONTROVERSY, GROWING POINTS, AND AREAS TIMELY FOR DEVELOPING RESEARCH: The design of clinical trials and the publication of unsatisfactory studies, in both human and animal models, continue to cause confusion in advising on patient management. Improvements in trial design, critical assessment of studies for publication and avoidance of bias towards publication of positive results are needed. A better understanding of pathogenesis should lead to more potent interventions. PMID- 20015854 TI - Of proverbs and prevention: aspiration and its consequences in older patients. PMID- 20015855 TI - The association between choice stepping reaction time and falls in older adults- a path analysis model. AB - BACKGROUND: choice stepping reaction time (CSRT) is a functional measure that has been shown to significantly discriminate older fallers from non-fallers. OBJECTIVE: to investigate how physiological and cognitive factors mediate the association between CSRT performance and multiple falls by use of path analysis. METHODS: 294 retirement-village residents, aged 62-95 years, undertook CSRT tests, requiring them to step onto one of four randomly illuminated panels, in addition to physiological and cognitive tests. Number of falls was collected during 1-year follow-up. RESULTS: 79 participants (27%) reported two or more falls during the follow-up period. Regression analyses indicated CSRT was able to predict multiple falls by a factor of 1.76 for each SD change. The path analysis model revealed that the association between CSRT and multiple falls was mediated entirely by the physiological parameters reaction time and balance (postural sway) performance. These two parameters were in turn mediated over a physiological path (by quadriceps strength and visual contrast sensitivity) and a cognitive path (cognitive processing). CONCLUSIONS: this study provides an example of how path analysis can reveal mediators for the association between a functional measure and falls. Our model identified inter-relationships (with relative weights) between physiological and cognitive factors, CSRT and multiple falls. PMID- 20015856 TI - DNA barcodes for 1/1000 of the animal kingdom. AB - This study reports DNA barcodes for more than 1300 Lepidoptera species from the eastern half of North America, establishing that 99.3 per cent of these species possess diagnostic barcode sequences. Intraspecific divergences averaged just 0.43 per cent among this assemblage, but most values were lower. The mean was elevated by deep barcode divergences (greater than 2%) in 5.1 per cent of the species, often involving the sympatric occurrence of two barcode clusters. A few of these cases have been analysed in detail, revealing species overlooked by the current taxonomic system. This study also provided a large-scale test of the extent of regional divergence in barcode sequences, indicating that geographical differentiation in the Lepidoptera of eastern North America is small, even when comparisons involve populations as much as 2800 km apart. The present results affirm that a highly effective system for the identification of Lepidoptera in this region can be built with few records per species because of the limited intra-specific variation. As most terrestrial and marine taxa are likely to possess a similar pattern of population structure, an effective DNA-based identification system can be developed with modest effort. PMID- 20015857 TI - Editorial 2010. PMID- 20015858 TI - Stable isotope views on ecosystem function: challenging or challenged? AB - Stable isotopes and their potential for detecting various and complex ecosystem processes are attracting an increasing number of scientists. Progress is challenging, particularly under global change scenarios, but some established views have been challenged. The IX meeting of the Spanish Association of Terrestrial Ecology (AAET, Ubeda, 18-22 October 2009) hosted a symposium on the ecology of stable isotopes where the linear mixing model approach of partitioning sinks and sources of carbon and water fluxes within an ecosystem was challenged, and new applications of stable isotopes for the study of plant interactions were evaluated. Discussion was also centred on the need for networks that monitor ecological processes using stable isotopes and key ideas for fostering future research with isotopes. PMID- 20015859 TI - Brain activity in an awake chimpanzee in response to the sound of her own name. AB - The brain activity of a fully awake chimpanzee being presented with her name was investigated. Event-related potentials (ERPs) were measured for each of the following auditory stimuli: the vocal sound of the subject's own name (SON), the vocal sound of a familiar name of another group member, the vocal sound of an unfamiliar name and a non-vocal sound. Some differences in ERP waveforms were detected between kinds of stimuli at latencies at which P3 and Nc components are typically observed in humans. Following stimulus onset, an Nc-like negative shift at approximately 500 ms latency was observed, particularly in response to SON. Such specific ERP patterns suggest that the chimpanzee processes her name differently from other sounds. PMID- 20015860 TI - A banned variety was the mother of several major wine grapes. AB - A number of widely grown varieties of Vitis vinifera ssp. sativa, the grape used for wine production, are known to have resulted from crosses between Pinot noir and Gouais blanc, although it is not known which was the maternal parent in these crosses. We have analysed microsatellites and a single nucleotide polymorphism (SNP) in chloroplast DNA from these two varieties and twelve progeny strains, including Chardonnay, Gamay noir and Aligote. The results demonstrate that Gouais blanc was the maternal parent for nine of these strains, including Chardonnay, Gamay noir and Aligote. This is a striking conclusion, as Gouais is generally considered a highly inferior variety, and its cultivation was banned for many years in parts of Europe. PMID- 20015861 TI - Evolution of ultraviolet vision in shorebirds (Charadriiformes). AB - Diurnal birds belong to one of two classes of colour vision. These are distinguished by the maximum absorbance wavelengths of the SWS1 visual pigment sensitive to violet (VS) and ultraviolet (UVS). Shifts between the classes have been rare events during avian evolution. Gulls (Laridae) are the only shorebirds (Charadriiformes) previously reported to have the UVS type of opsin, but too few species have been sampled to infer that gulls are unique among shorebirds or that Laridae is monomorphic for this trait. We have sequenced the SWS1 opsin gene in a broader sample of species. We confirm that cysteine in the key amino acid position 90, characteristic of the UVS class, has been conserved throughout gull evolution but also that the terns Anous minutus, A. tenuirostris and Gygis alba, and the skimmer Rynchops niger carry this trait. Terns, excluding Anous and Gygis, share the VS conferring serine in position 90 with other shorebirds but it is translated from a codon more similar to that found in UVS shorebirds. The most parsimonious interpretation of these findings, based on a molecular gene tree, is a single VS to UVS shift and a subsequent reversal in one lineage. PMID- 20015862 TI - Human papillomavirus type 16 E5 protein inhibits hydrogen-peroxide-induced apoptosis by stimulating ubiquitin-proteasome-mediated degradation of Bax in human cervical cancer cells. AB - To investigate the mechanism by which the human papillomavirus (HPV) E5 protein contributes to the carcinogenesis of uterine cervical cancer, we studied the effect of HPV E5 on apoptosis of cervical cancer cells and its underlying mechanism. Expression of HPV16 E5 protein inhibited hydrogen peroxide-induced apoptosis in C-33A cervical cancer cells. E5 decreased the expression of Bax protein, and exogenous expression of Bax abolished the anti-apoptotic effect of E5. Knockdown of E5 by small interfering RNA sensitized CaSki cervical cancer cells to hydrogen peroxide-induced apoptosis with concurrent increase in Bax expression. Transient expression of E5 significantly increased the degradation rate of Bax protein by inducing the ubiquitination. The E5-induced decrease in Bax expression was inhibited by a cyclooxygenase-2 (COX-2) inhibitor, prostaglandin E2 (PGE(2)) receptor antagonists and cyclic adenosine monophosphate dependent protein kinase (PKA) inhibitor. Treatment with PGE(2) decreased the expression of Bax and inhibited hydrogen peroxide-induced apoptosis of C-33A cells. We concluded that HPV16 E5 protein inhibits hydrogen peroxide-induced apoptosis of cervical cancer cells by stimulating the ubiquitin-proteasome mediated degradation of Bax protein, and the pathway involves COX-2, PGE(2) and PKA. This finding suggests the possibility that HPV 16 E5 protein contributes to cervical carcinogenesis by inhibiting apoptosis of transformed cervical epithelial cells. PMID- 20015863 TI - IQGAP1 and vimentin are key regulator genes in naturally occurring hepatotumorigenesis induced by oxidative stress. AB - To identify key genes involved in the complex multistep process of hepatotumorigenesis, we reduced multivariate clinicopathological variables by using the Long-Evans Cinnamon rat, a model with naturally occurring and oxidative stress-induced hepatotumorigenesis. Gene expression patterns were analyzed serially by profiling liver tissues from rats of a naive status (4 weeks old), through to those with chronic hepatitis (26 and 39 weeks old) to tumor development (67 weeks old). Of 31 099 probe sets used for microarray analysis, 87 were identified as being upregulated in a stepwise manner during disease progression and tumor development. Quantitative real-time reverse transcription polymerase chain reaction and statistical analyses verified that IQGAP1 and vimentin mRNA expression levels increased significantly throughout hepatotumorigenesis. A hierarchical clustering algorithm showed both genes clustered together and in the same cluster group. Immunohistochemical and western blot analyses showed similar increases in protein levels of IAGAP1 and vimentin. Finally, pathway analyses using text-mining technology with more comprehensive and recent gene-gene interaction data identified IQGAP1 and vimentin as important nodes in underlying gene regulatory networks. These findings enhance our understanding of the multistep hepatotumorigenesis and identification of target molecules for novel treatments. PMID- 20015864 TI - Downregulation of ANP32B, a novel substrate of caspase-3, enhances caspase-3 activation and apoptosis induction in myeloid leukemic cells. AB - The acidic leucine-rich nuclear phosphoprotein 32 (ANP32)B has been reported to regulate gene expression by acting as a histone chaperone or modulate messenger RNA trafficking by serving as a HuR ligand. However, its exact cellular functions are poorly understood. By utilizing a proteomics-based approach, in this work, we identify that the human ANP32B protein is cleaved during apoptosis induction by NSC606985, a novel camptothecin analog. Further investigation shows that various apoptosis inducers cause a decrease of full-length ANP32B in multiple cell lines with a concomitant increase of an approximately 17 kDa fragment. The proteolytic cleavage of ANP32B is inhibited by a specific caspase-3 inhibitor Z-DEVD-fmk, and it cannot be seen in NSC606985-induced death of caspase-3-deficient MCF-7 cells. In vitro caspase cleavage assay and mutagenesis experiment reveal that ANP32B is a direct substrate of caspase-3 and it is primarily cleaved at the sequence of Ala-Glu-Val-Asp, after Asp-163. Additionally, the reduced expression of endogenous ANP32B by specific small interfering RNA enhances caspase-3 activation and apoptosis induction by NSC606985 and etoposide. These results suggest that ANP32B is a novel substrate for caspase-3 and acts as a negative regulator for apoptosis, the mechanism of which remains to be explored. PMID- 20015865 TI - Gene expression profile in a glioma cell line resistant to cell death induced by the chimeric tumor suppressor-1 (CTS-1), a dominant-positive variant of p53--the role of NFkappaB. AB - The identification of genes involved in carcinogenesis and tumor progression is of great interest since these genes might be feasible as candidates for new tumor targeted therapy strategies. Chimeric tumor suppressor-1 (CTS-1), an artificial synthetic variant of p53, resists common p53 inactivation and could therefore be defined as a dominant-positive p53 variant. Overexpression of CTS-1 induces caspase-independent cell death. We used whole-genome microarray expression analysis in a parental (229(P)) and a CTS-1-resistant glioma cell line (229(Res)) to analyze alterations in gene expression in Ad-CTS-1-infected and in uninfected parental and resistant cells. In total, 700 genes were differentially expressed in infected and 313 genes in uninfected 229(Res) versus 229(P) cells. Ingenuity Pathway Analysis determined a variety of differentially expressed genes in Ad-CTS 1-infected cells that were members of intracellular networks with central tumor involved players such as nuclear factor-kappaB (NFkappaB), protein kinase B/AKT or transforming growth factor-beta. Here we focused on the function of NFkappaB in Ad-CTS-1-mediated cell death in glioma. NFkappaB was activated in Ad-CTS-1 infected 229(P) but not 229(Res) cells. NFkappaB activation was accompanied by the induction of cell death in parental cells. Inhibition of NFkappaB activity by expression of an IkappaB super repressor or upregulation of the NFkappaB-linked gene Bex protected parental cells to Ad-CTS-1-induced cell death, whereas knockdown of Bex sensitized both parental and resistant cells. Taken together, these data suggest that activation of the normally antiapoptotic protein NFkappaB does not always necessarily protect cells from apoptosis but, in the glioma cell lines tested so far, and in an environment where p53 is constitutively active, also leads to the induction of cell death. PMID- 20015866 TI - Polymerization by DNA polymerase eta is blocked by cis diamminedichloroplatinum(II) 1,3-d(GpTpG) cross-link: implications for cytotoxic effects in nucleotide excision repair-negative tumor cells. AB - cis-Diamminedichloroplatinum(II) (cisplatin) forms DNA adducts that interfere with replication and transcription. The most common adducts formed in vivo are 1,2-intrastrand d(GpG) cross-links (Pt-GG) and d(ApG) cross-links (Pt-AG), with minor amounts of 1,3-d(GpNpG) cross-links (Pt-GNG), interstrand cross-links and monoadducts. Although the relative contribution of these different adducts to toxicity is not known, literature implicates that Pt-GG and Pt-AG adducts block replication. Thus, nucleotide excision repair (NER), by which platinum adducts are excised, and translesion DNA synthesis (TLS), which permits adduct bypass, are thought to be associated with cisplatin resistance. Recent studies have reported that the clinical benefit from platinum-based chemotherapy is high if tumor cells express low levels of NER factors. To investigate the role of platinum-DNA adducts in mediating tumor cell survival by TLS, we examined whether 1,3-intrastrand d(GpTpG) platinum cross-links (Pt-GTG), which probably exist in NER-negative tumor cells but not in NER-positive tumor cells, are bypassed by the translesion DNA polymerase eta (pol eta), which is known to bypass Pt-GG. We show that pol eta can incorporate the correct deoxycytidine triphosphate opposite the first 3'-cross-linked G of Pt-GTG but cannot insert any nucleotides opposite the second intact T or the third 5'-cross-linked G of the adducts, thereby suggesting that TLS does not facilitate replication past Pt-GTG adducts. Thus, our findings implicate Pt-GNG adducts as mediating the cytotoxicity of platinum-DNA adducts in NER-negative tumors in vivo. PMID- 20015868 TI - Determinants of participation in colorectal cancer screening with faecal occult blood testing. AB - BACKGROUND: Colorectal cancer is one of the most common cancers in men and women. Participation rates in faecal occult blood testing (FOBT) screening activities are, however, relatively low. In terms of lowering the colorectal cancer mortality, high participation rates are essential, and therefore it is important to understand the barriers to FOBT screening. METHODS: We undertook a systematic search through PUBMED, Medline, EMBASE and PsycINFO in order to identify studies that provide information on socio-demographic determinants of participation in FOBT screening. RESULTS: FOBT participation varied considerably across countries, but they have rarely been above 60%. The use of other health-care services was in most studies a strong determinant for participation in screening with FOBT. There was a tendency to higher participation among women than among men and among married as opposed to not married, but determinants varied across countries and test settings. There was no systematic variation in participation across age groups. CONCLUSION: The participation pattern depends in part on local circumstances, which makes it difficult to point to a general strategy for increasing the uptake in FOBT screening. This stresses the need for monitoring of individual screening programmes and developing information strategies targeted to the local participation pattern. PMID- 20015867 TI - The Bmi-1 polycomb protein antagonizes the (-)-epigallocatechin-3-gallate dependent suppression of skin cancer cell survival. AB - The polycomb group (PcG) proteins are epigenetic regulators of gene expression that enhance cell survival. This regulation is achieved via action of two multiprotein PcG complexes--PRC2 (EED) and PRC1 [B-cell-specific Moloney murine leukemia virus integration site 1 (Bmi-1)]. These complexes modulate gene expression by increasing histone methylation and reducing acetylation--leading to a closed chromatin conformation. Activity of these proteins is associated with increased cell proliferation and survival. We show increased expression of key PcG proteins in immortalized keratinocytes and skin cancer cell lines. We examine the role of two key PcG proteins, Bmi-1 and enhancer of zeste homolog 2 (Ezh2), and the impact of the active agent in green tea, (-)-epigallocatechin-3-gallate (EGCG), on the function of these regulators. EGCG treatment of SCC-13 cells reduces Bmi-1 and Ezh2 level and this is associated with reduced cell survival. The reduction in survival is associated with a global reduction in histone H3 lysine 27 trimethylation, a hallmark of PRC2 complex action. This change in PcG protein expression is associated with reduced expression of key proteins that enhance progression through the cell cycle [cyclin-dependent kinase (cdk)1, cdk2, cdk4, cyclin D1, cyclin E, cyclin A and cyclin B1] and increased expression of proteins that inhibit cell cycle progression (p21 and p27). Apoptosis is also enhanced, as evidenced by increased caspase 9, 8 and 3 cleavage and increased poly(adenosine diphosphate ribose) polymerase cleavage. EGCG treatment also increases Bax and suppresses Bcl-xL expression. Vector-mediated enhanced Bmi-1 expression reverses these EGCG-dependent changes. These findings suggest that green tea polyphenols reduce skin tumor cell survival by influencing PcG-mediated epigenetic regulatory mechanisms. PMID- 20015869 TI - Production of viable piglets for the first time using sperm derived from ectopic testicular xenografts. AB - Xenografting of testicular tissue into immunodeficient mice is known to be a valuable tool for facilitating the development of immature germ cells present in mammalian gonads. Spermatogenesis in xenografts and/or in vitro embryonic development to the blastocyst stage after ICSI of xenogeneic sperm has already been reported in large animals, including pigs; however, development of the embryos to term has not yet been confirmed. Therefore, in pigs, we evaluated the in vivo developmental ability of oocytes injected after ICSI of xenogeneic sperm. Testicular tissues prepared from neonatal piglets, which contain seminiferous cords consisting of only gonocytes/spermatogonia, were transplanted under the back skin of castrated nude mice. Between 133 and 280 days after xenografting, morphologically normal sperm were recovered, and a single spermatozoon was then injected into an in vitro matured porcine oocyte. After ICSI, the oocytes were electrostimulated and transferred into estrus-synchronized recipients. Two out of 23 recipient gilts gave birth to six piglets. Here, we describe for the first time that oocytes fertilized with a sperm from ectopic xenografts have the ability to develop to viable offspring in large mammals. PMID- 20015870 TI - Physiological and glycomic characterization of N-acetylglucosaminyltransferase IVa and -IVb double deficient mice. AB - N-Acetylglucosaminyltransferase-IV (GnT-IV) has two isoenzymes, GnT-IVa and GnT IVb, which initiate the GlcNAcbeta1-4 branch synthesis on the Manalpha1-3 arm of the N-glycan core thereby increasing N-glycan branch complexity and conferring endogenous lectin binding epitopes. To elucidate the physiological significance of GnT-IV, we engineered and characterized GnT-IVb-deficient mice and further generated GnT-IVa/-IVb double deficient mice. In wild-type mice, GnT-IVa expression is restricted to gastrointestinal tissues, whereas GnT-IVb is broadly expressed among organs. GnT-IVb deficiency induced aberrant GnT-IVa expression corresponding to the GnT-IVb distribution pattern that might be attributed to increased Ets-1, which conceivably activates the Mgat4a promoter, and thereafter preserved apparent GnT-IV activity. The compensative GnT-IVa expression might contribute to amelioration of the GnT-IVb-deficient phenotype. GnT-IVb deficiency showed mild phenotypic alterations in hematopoietic cell populations and hemostasis. GnT-IVa/-IVb double deficiency completely abolished GnT-IV activity that resulted in the disappearance of the GlcNAcbeta1-4 branch on the Manalpha1-3 arm that was confirmed by MALDI-TOF MS and GC-MS linkage analyses. Comprehensive glycomic analyses revealed that the abundance of terminal moieties was preserved in GnT-IVa/-IVb double deficiency that was due to the elevated expression of glycosyltransferases regarding synthesis of terminal moieties. Thereby, this may maintain the expression of glycan ligands for endogenous lectins and prevent cellular dysfunctions. The fact that the phenotype of GnT-IVa/-IVb double deficiency largely overlapped that of GnT-IVa single deficiency can be attributed to the induced glycomic compensation. This is the first report that mammalian organs have highly organized glycomic compensation systems to preserve N-glycan branch complexity. PMID- 20015872 TI - A novel XIAP mutation in a Japanese boy with recurrent pancytopenia and splenomegaly. PMID- 20015871 TI - Pharmacogenetic risk factors for altered bone mineral density and body composition in pediatric acute lymphoblastic leukemia. AB - BACKGROUND: This study investigates pharmacogenetic risk factors for bone mineral (apparent) density (BM(A)D) and body composition in pediatric acute lymphoblastic leukemia DESIGN AND METHODS: We determined the influence of SNPs in 4 genes (vitamin-D receptor (VDR: BsmI/ApaI/TaqI and Cdx-2/GATA), collagen type I alpha 1 (SpI), estrogen receptor 1 (ESR1: PvuII/XbaI), glucocorticoid receptor (BclI)) on body composition, BM(A)D and fracture risk during dexamethasone-based pediatric acute lymphoblastic leukemia treatment. Body composition and BMD were measured repeatedly during and after treatment using dual energy X-ray absorptiometry. RESULTS: Non-carriers of VDR 5'-end (Cdx-2/GATA) haplotype 3 revealed a significant larger fat gain than carriers (Delta%fat: non-carriers: +1.76SDS, carriers: +0.77SDS, P<0.001). At diagnosis and during therapy, lumbar spine BMD was significantly higher in non-carriers of VDR 5'-end (Cdx-2/GATA) haplotype 3 than in carriers. The other SNPs did not influence BMD or fracture risk during/after treatment. The year after treatment completion, lean body mass increased in non-carriers of ESR1 (PvuII/XbaI) haplotype 3 and decreased in carriers (Delta lean body mass: non-carriers:+0.28SDS, carriers: -0.55SDS, P<0.01). CONCLUSIONS: Only VDR 5'-end (Cdx-2/GATA) haplotype 3 was identified as protective factor against excessive fat gain and as a risk factor for lower lumbar spine BMD during treatment. Carrying ESR1 (PvuII/XbaI) haplotype 3 negatively influenced recovery of lean body mass after pediatric acute lymphoblastic leukemia treatment. PMID- 20015873 TI - Intercellular adhesion molecule-4 and CD36 are implicated in the abnormal adhesiveness of sickle cell SAD mouse erythrocytes to endothelium. AB - BACKGROUND: Abnormal adhesiveness of red blood cells to endothelium has been implicated in vaso-occlusive crisis of sickle cell disease. The present study examined whether the SAD mouse model exhibits the same abnormalities of red blood cell adhesion as those found in human sickle cell disease. DESIGN AND METHODS: The repertoire of adhesive molecules on murine erythrocytes and bEnd.3 microvascular endothelial cells was determined by flow cytometry using monoclonal antibodies or by western blotting. Adhesion was investigated in dynamic conditions and measured at different shear stresses. RESULTS: CD36, CD47 and intercellular adhesion molecular-4, but not Lutheran blood group antigen/basal cell adhesion molecule, are present on mouse mature erythrocytes. alpha(4)beta(1) are not expressed on SAD and wild type reticulocytes. Endothelial bEnd.3 cells express alpha(V)beta(3), alpha(4)beta(1), CD47, vascular cell adhesion molecule 1, and Lutheran blood group antigen/basal cell adhesion molecule, but not CD36. Adhesion of SAD red cells is: (i) 2- to 3-fold higher than that of wild type red cells; (ii) further increased on platelet activating factor-activated endothelium; (iii) not stimulated by epinephrine; (iv) inhibited after treating the endothelium with a peptide reproducing one of the binding sequences of mouse intercellular adhesion molecular-4, or with mon-oclonal antibody against murine alpha(v) integrin; and (v) inhibited after pretreatment of red blood cells with anti-mouse CD36 monoclonal antibodies. The combination of treatments with intercellular adhesion molecular-4 peptide and anti-CD36 monoclonal antibodies eliminates excess adhesion of SAD red cells. The phosphorylation state of intercellular adhesion molecular-4 and CD36 is probably not involved in the over adhesiveness of SAD erythrocytes. CONCLUSIONS: Intercellular adhesion molecular 4/alpha(v)beta(3) and CD36/thrombospondin interactions might contribute to the abnormally high adhesiveness of SAD red cells. The SAD mouse is a valuable animal model for investigating adhesion processes of sickle cell disease. PMID- 20015874 TI - Myelin protein zero is naturally processed in the B cells of monoclonal gammopathy of undetermined significance of immunoglobulin M isotype: aberrant triggering of a patient's T cells. AB - BACKGROUND: Monoclonal gammopathy of undetermined significance of immunoglobulin M isotype is a condition with clonally expanded B cells, recently suggested to have an infectious origin. This monoclonal gammopathy is frequently associated with polyneuropathy and antibodies against myelin protein zero, whereas the role of the T cells remains largely unknown. We analyzed protein zero-specific B cells, as antigen-presenting cells, and their capacity to activate T helper cells. DESIGN AND METHODS: We used a well-characterized monoclonal gammopathy of undetermined significance-derived B-cell line, TJ2, expressing anti-protein zero immunoglobulin M. The ability of TJ2 cells to bind, endocytose, process, and present protein zero was investigated by receptor-clustering and immunofluorescence. The activation of protein zero-specific autologous T cells was studied by measuring interleukin-2 and interferon-gamma with flow cytometry, immunobeads, and enzyme-linked immunospot assays. RESULTS: Surface-receptor clustering and endocytosis of receptor-ligand (immunoglobulin M/protein zero) complexes were pronounced after exposure to protein zero. Naturally processed or synthetic protein zero peptide (194-208)-pulsed TJ2 cells significantly induced interleukin-2 secretion from autologous T cells compared to control antigen pulsed cells (P<0.001). The numbers of interferon-gamma-producing T helper cells, including CD4(+)/CD8(+) cells, were also significantly increased (P=0.0152). Affinity-isolated naturally processed myelin peptides were potent interferon gamma stimulators for autologous peripheral blood mononuclear cells, but not for control peripheral blood mononuclear cells. CONCLUSIONS: We show for the first time that myelin protein zero is naturally processed in B cells from monoclonal gammopathy of undetermined significance of immunoglobulin M isotype, acting as aberrant antigen-presenting cells in activation of a patient's T helper cells. Our findings cast new light on the important role of autoreactive protein zero specific B cells in the induction of the pathogenic T-cell responses found in nerve lesions of patients with monoclonal gammopathy of undetermined significance with peripheral neuropathy. PMID- 20015875 TI - Early hemorrhagic death before starting therapy in acute promyelocytic leukemia: association with high WBC count, late diagnosis and delayed treatment initiation. PMID- 20015876 TI - Meta-analysis on hypomethylating agents in myelodysplastic syndromes. PMID- 20015877 TI - C/EBPbeta expression in ALK-positive anaplastic large cell lymphomas is required for cell proliferation and is induced by the STAT3 signaling pathway. AB - BACKGROUND: Anaplastic lymphoma kinase (ALK)-positive anaplastic large cell lymphoma is characterized by the t(2;5) chromosomal translocation, resulting in the expression of a fusion protein formed of nucleophosmin (NPM) and ALK. Recently, we reported the abnormal expression of the transcription factor CCAAT/enhancer binding protein-beta (C/EBPbeta) in ALK-positive anaplastic large cell lymphomas, and demonstrated its dependence on NPM-ALK activity. DESIGN AND METHODS: In this study, the role of C/EBPbeta in proliferation and survival of ALK-positive anaplastic large cell lymphomas was investigated, as well as the mechanism of its expression and activity. Highly effective short hairpin RNA sequences and/or pharmacological inhibitors were used to abrogate the expression or activity of C/EBPbeta, signal transducer and activator of transcription 3 (STAT3), AKT, extracellular signal-related kinase 1/2 (ERK1/2) and mammalian target of rapamycin (mTOR). RESULTS: Interference with C/EBPbeta expression resulted in a dramatic decrease in cell proliferation in ALK-positive anaplastic large cell lymphomas, with a mild induction of apoptosis after 6 days. Down regulation of STAT3 resulted in a marked decrease in C/EBPbeta mRNA and protein levels with impairment in cell proliferation and viability, underscoring the important role of these two proteins in ALK-mediated oncogenesis. Additionally, we demonstrated that reduction of ERK1/2 activity led to C/EBPbeta Thr(235) dephosphorylation and moderate growth retardation. The AKT/mTOR signaling pathway did not have any influence on C/EBPbeta expression or C/EBPbeta phosphorylation. CONCLUSIONS: These findings reveal the convergence of STAT3 and ERK1/2 signaling pathways activated by NPM-ALK in mediating the regulation of C/EBPbeta expression, a transcription factor central to NPM-ALK transformation. PMID- 20015878 TI - Hypoxia-inducible factor-2 is a novel regulator of aberrant CXCL12 expression in multiple myeloma plasma cells. AB - BACKGROUND: Multiple myeloma is an incurable malignancy of bone marrow plasma cells. Progression of multiple myeloma is accompanied by an increase in bone marrow angiogenesis. Studies from our laboratory suggest a role for the CXCL12 chemokine in this process, with circulating levels of CXCL12 correlating with bone marrow angiogenesis in patients with multiple myeloma. While the mechanisms responsible for aberrant plasma cell expression of CXCL12 remain to be determined, studies in other systems suggest a role for hypoxia and hypoxia inducible transcription factors. DESIGN AND METHODS: The expression of hypoxia inducible factor protein was examined in patients' bone marrow biopsy specimens using immunohistochemistry. The hypoxic regulation of CXCL12 was examined in multiple myeloma plasma cell lines using polymerase chain reaction and western blotting. The role of hypoxia-inducible factors-1 and -2 in the regulation of CXCL12 expression was examined using over-expression and short hairpin RNA knockdown constructs, electrophoretic mobility shift assays and chromatin immunoprecipitation. The contribution of CXCL12 to hypoxia-induced angiogenesis was examined in vivo using a subcutaneous murine model of neovascularization. RESULTS: Strong hypoxia-inducible factor-2 protein expression was detected in CD138(+) multiple myeloma plasma cells in patients' biopsy specimens. Prolonged exposure to hypoxia strongly up-regulated CXCL12 expression in multiple myeloma plasma cells and hypoxia-inducible factor-2 was found to play a key role in this response. Promoter analyses revealed increased hypoxia-inducible factor-2 binding to the CXCL12 promoter under hypoxic conditions. Over-expression of hypoxia inducible factor in multiple myeloma plasma cells strongly induced in vivo angiogenesis, and administration of a CXCL12 antagonist decreased hypoxia inducible factor-induced angiogenesis. CONCLUSIONS: Hypoxia-inducible factor-2 is a newly identified regulator of CXCL12 expression in multiple myeloma plasma cells and a major contributor to multiple myeloma plasma cell-induced angiogenesis. Targeting the hypoxic niche, and more specifically hypoxia inducible factor-2, may represent a viable strategy to inhibit angiogenesis in multiple myeloma and progression of this disease. PMID- 20015879 TI - Cryohydrocytosis: increased activity of cation carriers in red cells from a patient with a band 3 mutation. AB - BACKGROUND: Cryohydrocytosis is an inherited dominant hemolytic anemia characterized by mutations in a transmembrane segment of the anion exchanger (band 3 protein). Transfection experiments performed in Xenopus oocytes suggested that these mutations may convert the anion exchanger into a non-selective cation channel. The present study was performed to characterize so far unexplored ion transport pathways that may render erythrocytes of a single cryohydrocytosis patient cation-leaky. DESIGN AND METHODS: Cold-induced changes in cell volume were monitored using ektacytometry and density gradient centrifugation. Kinetics, temperature and inhibitor-dependence of the cation and water movements in the cryohydrocytosis patient's erythrocytes were studied using radioactive tracers and flame photometry. Response of the membrane potential of the patient's erythrocyte membrane to the presence of ionophores and blockers of anion and cation channels was assessed. RESULTS: In the cold, the cryohydrocytosis patient's erythrocytes swelled in KCl-containing, but not in NaCl-containing or KNO(3)-containing media indicating that volume changes were mediated by an anion coupled cation transporter. In NaCl-containing medium the net HOE-642-sensitive Na(+)/K(+) exchange prevailed, whereas in KCl-containing medium swelling was mediated by a chloride-dependent K(+) uptake. Unidirectional K(+) influx measurements showed that the patient's cells have abnormally high activities of the cation-proton exchanger and the K(+),Cl(-) co-transporter, which can account for the observed net movements of cations. Finally, neither chloride nor cation conductance in the patient's erythrocytes differed from that of healthy donors. Conclusions These results suggest that cross-talk between the mutated band 3 and other transporters might increase the cation permeability in cryohydrocytosis. PMID- 20015880 TI - Regulation of PTEN by CK2 and Notch1 in primary T-cell acute lymphoblastic leukemia: rationale for combined use of CK2- and gamma-secretase inhibitors. AB - T-cell acute lymphoblastic leukemia (T-ALL) patients frequently display NOTCH1 activating mutations and Notch can transcriptionally down-regulate the tumor suppressor PTEN. However, it is not clear whether NOTCH1 mutations associate with decreased PTEN expression in primary T-ALL. Here, we compared patients with or without NOTCH1 mutations and report that the former presented higher MYC transcript levels and decreased PTEN mRNA expression. We recently showed that T ALL cells frequently display CK2-mediated PTEN phosphorylation, resulting in PTEN protein stabilization and concomitant functional inactivation. Accordingly, the T ALL samples analyzed, irrespectively of their NOTCH1 mutational status, expressed significantly higher PTEN protein levels than normal controls. To evaluate the integrated functional impact of Notch transcriptional and CK2 post-translational inactivation of PTEN, we treated T-ALL cells with both the gamma-secretase inhibitor DAPT and the CK2 inhibitors DRB/TBB. Our data suggest that combined use of gamma-secretase and CK2 inhibitors may have therapeutic potential in T-ALL. PMID- 20015881 TI - Can antigen-specific regulatory T cells protect against graft versus host disease and spare anti-malignancy alloresponse? AB - Allogeneic hematopoietic cell transplantation provides effective control of hematopoietic malignancies, but with an associated risk of graft-versus-host disease (GVHD) related morbidity and mortality. Several advances in hematopoietic cell transplantation including high resolution HLA typing, development of reduced intensity conditioning regimens, infectious prophylaxis and treatment, and novel immunosuppressive agents have resulted in improved outcomes and improved access to transplantation, but GVHD remains a major obstacle. This clinico-pathological syndrome, mediated by donor alloreactive T cells, occurs often despite prophylactic immunosuppressive therapy. Regulatory T cells, a suppressive subset of the T-cell repertoire, may offer promise as a novel cellular therapy for more effective prevention of GVHD. While advances have been made in pre-clinical experimental animals, several challenges remain in the translation of this work to human trials. Strategies to effectively produce ex vivo expanded alloantigen specific regulatory T cells specific for ubiquitous alloantigens but sparing hematopoietic- or tumor-associated antigens hold promise to prevent GVHD while allowing a preserved graft versus malignancy effect. PMID- 20015882 TI - High-resolution single-nucleotide polymorphism array-profiling in myeloproliferative neoplasms identifies novel genomic aberrations. AB - Single-nucleotide polymorphism arrays allow for genome-wide profiling of copy number alterations and copy-neutral runs of homozygosity at high resolution. To identify novel genetic lesions in myeloproliferative neoplasms, a large series of 151 clinically well characterized patients was analyzed in our study. Copy-number alterations were rare in essential thrombocythemia and polycythemia vera. In contrast, approximately one third of myelofibrosis patients exhibited small genomic losses (less than 5 Mb). In 2 secondary myelofibrosis cases the tumor suppressor gene NF1 in 17q11.2 was affected. Sequencing analyses revealed a mutation in the remaining NF1 allele of one patient. In terms of copy-neutral aberrations, no chromosomes other than 9p were recurrently affected. In conclusion, novel genomic aberrations were identified in our study, in particular in patients with myelofibrosis. Further analyses on single-gene level are necessary to uncover the mechanisms that are involved in the pathogenesis of myeloproliferative neoplasms. PMID- 20015883 TI - Can cytoplasmic nucleophosmin be detected by immunocytochemical staining of cell smears in acute myeloid leukemia? AB - Mutations in the C-terminal region of nucleophosmin in acute myeloid leukemia (AML) result in aberrant cytoplasmic nucleophosmin (cNPM) in leukemic blast cells which is detectable by immunocytochemistry in bone marrow trephine (BMT) biopsy sections. We tested whether cNPM is detectable by immunocytochemistry in air dried smears of AML with nucleophosmin1 (NPM1) mutations. An immunoalkaline phosphatase method was developed using the OCI-AML3 cell line, known to have mutated NPM1, and assessed on blood and marrow smears of 60 AML cases. NPM was detectable in all blast cell nucleoli and cNPM in 21 of 31 of NPM1 mutated and 15 of 29 wild-type cases. Paired air-dried smears and BMT biopsies from the same case (mutated and wild-type) gave discrepancies in cNPM expression and there was no correlation in 10 of 22 cases. Due to the high false positive and negative rates for cNPM in cell smears, this method should not be used as a surrogate for NPM1 mutations in AML. PMID- 20015884 TI - Impact of additional chromosomal aberrations and BCR-ABL kinase domain mutations on the response to nilotinib in Philadelphia chromosome-positive chronic myeloid leukemia. AB - BACKGROUND: Additional chromosomal aberrations in Philadelphia chromosome positive chronic myeloid leukemia are non-random and strongly associated with disease progression, but their prognostic impact and effect on treatment response is not clear. Point mutations in the BCR-ABL kinase domain are probably the most common mechanisms of imatinib resistance. DESIGN AND METHODS: We assessed the influence of additional chromosomal aberrations and BCR-ABL kinase domain mutations on the response to the second-generation tyrosine kinase inhibitor nilotinib after imatinib-failure. Standard cytogenetic analysis of metaphases was performed to detect additional chromosomal aberrations and the BCR-ABL kinase domain was sequenced to detect point mutations. RESULTS: Among 53 patients with a median follow-up of 16 months, of whom 38, 5 and 10 were in chronic phase, accelerated phase and blast crisis, respectively, 19 (36%) had additional chromosomal aberrations and 20 (38%) had BCR-ABL kinase domain mutations. The 2 year overall survival rate of all patients with-out additional chromosomal aberrations (89%) was higher than that of patients with such aberrations (54%) (P=0.0025). Among patients with chronic phase disease, overall survival at 2 years was 100% and 62% for patients without or with additional chromosomal aberrations, respectively (P=0.0024). BCR-ABL kinase domain mutations were associated with lower remission rates in response to nilotinib, with 9 of 20 (45%) of these patients achieving a major cytogenetic remission as compared to 26 of 33 (79%) patients without mutations (P<0.05). However, overall survival was not affected by BCR-ABL kinase domain mutations. CONCLUSIONS: Whereas BCR-ABL kinase domain mutations may confer more specific resistance to nilotinib, which will predominantly affect response rates, the presence of additional chromosomal aberrations may reflect genetic instability and, therefore, intrinsic aggressiveness of the disease which will be less amenable to subsequent alternative treatments and thus negatively affect overall survival. Conventional cytogenetic analyses remain mandatory during follow-up of patients with chronic myeloid leukemia under tyrosine kinase inhibitor therapy. PMID- 20015885 TI - Cancer-testis antigens MAGE-C1/CT7 and MAGE-A3 promote the survival of multiple myeloma cells. AB - BACKGROUND: Multiple myeloma is a life-threatening disease and despite the introduction of stem cell transplantation and novel agents such as thalidomide, lenalidomide, and bortezomib most patients will relapse and develop chemoresistant disease. Therefore, alternative therapeutic modes for myeloma are needed and cancer-testis antigens such as MAGE-C1/CT7 and MAGE-A3 have been suggested to represent a class of tumor-specific proteins particularly suited for targeted immunotherapies. Surprisingly, the biological role of cancer-testis genes in myeloma remains poorly understood. DESIGN AND METHODS: We performed the first investigation of the function of two cancer-testis antigens most commonly expressed in myeloma, MAGE-C1/CT7 and MAGE-A3, using an RNA interference-based gene silencing model in myeloma cell lines. Functional assays were used to determine changes in proliferation, cell adhesion, chemosensitivity, colony formation, and apoptosis resulting from gene-specific silencing. RESULTS: We show that the investigated genes are not involved in regulating cell proliferation or adhesion; however, they play an important role in promoting the survival of myeloma cells. Accordingly, knock-down of MAGE-C1/CT7 and MAGE-A3 led to the induction of apoptosis in the malignant plasma cells and, importantly, both genes were also essential for the survival of clonogenic myeloma precursors. Finally, silencing of cancer-testis genes further improved the response of myeloma cells to conventional therapies. CONCLUSIONS: Cancer-testis antigens such as MAGE C1/CT7 and MAGE-A3 play an important role in promoting the survival of myeloma cells and clonogenic precursors by reducing the rate of spontaneous and chemotherapy-induced apoptosis and might, therefore, represent attractive targets for novel myeloma-specific therapies. PMID- 20015887 TI - CD5 expression identifies a subset of splenic marginal zone lymphomas with higher lymphocytosis: a clinico-pathological, cytogenetic and molecular study of 24 cases. AB - BACKGROUND: Classically, splenic marginal zone B-cell lymphoma is characterized by the absence of CD5 expression. Cases of apparent splenic marginal zone B-cell lymphoma showing CD5 expression, as diagnosed by blood studies, have been described; however, in the absence of histological evidence, the correct diagnosis of these cases is controversial because of possible confusion with other CD5-positive small B-cell neoplasms. DESIGN AND METHODS: We report a series of 24 CD5-positive, t(11;14)-negative cases of splenic marginal zone B-cell lymphoma diagnosed by flow cytometry studies of blood and histologically proven on spleen sections. Clinical data as well as morphological, immunological, cytogenetic and molecular characteristics were assessed to evaluate the similarities and differences of these cases with those of classical CD5-negative splenic marginal zone B-cell lymphoma. RESULTS: The CD5 expression detected in blood by flow cytometry was confirmed in most cases by immunohistochemistry on spleen sections. In general, cases of CD5-positive and CD5-negative splenic marginal zone B-cell lymphoma did not appear different and, in particular, they showed similar karyotypic changes such as 7q deletion, trisomy 3, trisomy 18 and biased IGHV usage (i.e. VH1-2). The main differences were a higher lymphocyte count at diagnosis (8.15x10(9)/L versus 3.90x10(9)/L; P=0.005) and more frequent diffuse bone marrow infiltration (34% versus 8%; P=0.03) in the CD5-positive group. A tendency to a more mutated IGHV status in the CD5 positive cases was observed (80% versus 54.5%; (P=0.11). No significant differences in outcome were found in relation to CD5 expression. CONCLUSIONS: This study confirms the existence of cases of CD5-positive splenic marginal zone B-cell lymphoma and shows that these cases are closely related to classical splenic marginal zone lymphoma. Whether or not CD5-positive splenic marginal zone B-cell lymphoma constitutes a true subset obviously requires the study of more cases. PMID- 20015886 TI - Efficacy of escalated imatinib combined with cytarabine in newly diagnosed patients with chronic myeloid leukemia. AB - BACKGROUND: In order to improve the molecular response rate and prevent resistance to treatment, combination therapy with different dosages of imatinib and cytarabine was studied in newly diagnosed patients with chronic myeloid leukemia in the HOVON-51 study. DESIGN AND METHODS: Having reported feasibility previously, we hereby report the efficacy of escalated imatinib (200 mg, 400 mg, 600 mg or 800 mg) in combination with two cycles of intravenous cytarabine (200 mg/m(2) or 1000 mg/m(2) days 1 to 7) in 162 patients with chronic myeloid leukemia. RESULTS: With a median follow-up of 55 months, the 5-year cumulative incidences of complete cytogenetic response, major molecular response, and complete molecular response were 89%, 71%, and 53%, respectively. A higher Sokal risk score was inversely associated with complete cytogenetic response (hazard ratio of 0.63; 95% confidence interval, 0.50-0.79, P<0.001). A higher dose of imatinib and a higher dose of cytarabine were associated with increased complete molecular response with hazard ratios of 1.60 (95% confidence interval, 0.96 2.68, P=0.07) and 1.66 (95% confidence interval, 1.02-2.72, P=0.04), respectively. Progression-free survival and overall survival rates at 5 years were 92% and 96%, respectively. Achieving a major molecular response at 1 year was associated with complete absence of progression and a probability of achieving a complete molecular response of 89%. CONCLUSIONS: The addition of intravenous cytarabine to imatinib as upfront therapy for patients with chronic myeloid leukemia is associated with a high rate of complete molecular responses (Clinicaltrials.Gov Identifier: NCT00028847). PMID- 20015888 TI - UNC13D is the predominant causative gene with recurrent splicing mutations in Korean patients with familial hemophagocytic lymphohistiocytosis. AB - BACKGROUND: Familial hemophagocytic lymphohistiocytosis is a fatal disease characterized by immune dysregulation from defective function of cytotoxic lymphocytes. Three causative genes have been identified for this autosomal recessive disorder (PRF1, UNC13D, and STX11). We investigated the molecular genetics of familial hemophagocytic lymphohistiocytosis in Korea. DESIGN AND METHODS: Pediatric patients who fulfilled the HLH-2004 criteria were recruited from the Korean Registry for Histiocytosis. Molecular genetic studies were performed on the patients' DNA samples by direct sequencing of all coding exons and flanking sequences of PRF1, UNC13D, and STX11. RESULTS: Forty patients were studied and familial hemophagocytic lymphohistiocytosis mutations were identified in nine; eight patients had UNC13D mutations (89%) and one had a mutation in PRF1. No patient had a STX11 mutation. Notably, four patients had only one UNC13D mutant allele, suggesting that the other mutation was missed by conventional direct sequencing. All UNC13D mutations were deleterious in nature. One known splicing mutation, c.754-1G>C, was recurrent, accounting for 58% of all the mutant alleles (7/12). Five UNC13D mutations were novel (p.Gln98X, p.Glu565SerfsX7, c.1993-2A>G, c.2367+1G>A, and c.2954+5G>A). The one patient with PRF1 mutation was homozygous for a frameshift mutation (p.Leu364GlufsX93), which was previously reported to be the most frequent PRF1 mutation in Japan. CONCLUSIONS: This is the first investigation on the molecular genetics of familial hemophagocytic lymphohistiocytosis in Korea. The data showed that UNC13D is the predominant causative gene in the Korean population. The identification of mutations missed by conventional sequencing would better delineate the mutation spectrum and help to establish the optimal molecular diagnostic strategy for familial hemophagocytic lymphohistiocytosis in Korea, which might need an RNA based screening strategy. PMID- 20015889 TI - Mouse mesenchymal stem cells can support human hematopoiesis both in vitro and in vivo: the crucial role of neural cell adhesion molecule. AB - BACKGROUND: We previously established a mesenchymal stem cell line (FMS/PA6-P) from the bone marrow adherent cells of fetal mice. The cell line expresses a higher level of neural cell adhesion molecule and shows greater hematopoiesis supporting capacity in mice than other murine stromal cell lines. DESIGN AND METHODS: Since there is 94% homology between human and murine neural cell adhesion molecule, we examined whether FMS/PA6-P cells support human hematopoiesis and whether neural cell adhesion molecules expressed on FMS/PA6-P cells contribute greatly to the human hematopoiesis-supporting ability of the cell line. RESULTS: When lineage-negative cord blood mononuclear cells were co cultured on the FMS/PA6-P cells, a significantly greater hematopoietic stem cell enriched population (CD34(+)CD38(-) cells) was obtained than in the culture without the FMS/PA6-P cells. Moreover, when lineage-negative cord blood mononuclear cells were cultured on FMS/PA6-P cells and transplanted into SCID mice, a significantly larger proportion of human CD45(+) cells and CD34(+)CD38(-) cells were detected in the bone marrow of SCID mice than in the bone marrow of SCID mice that had received lineage-negative cord blood mononuclear cells cultured without FMS/PA6-P cells. Furthermore, we found that direct cell-to-cell contact between the lineage-negative cord blood mononuclear cells and the FMS/PA6 P cells was essential for the maximum expansion of the mononuclear cells. The addition of anti-mouse neural cell adhesion molecule antibody to the culture significantly inhibited their contact and the proliferation of lineage-negative cord blood mononuclear cells. CONCLUSIONS: These findings suggest that neural cell adhesion molecules expressed on FMS/PA6-P cells play a crucial role in the human hematopoiesis-supporting ability of the cell line. PMID- 20015891 TI - A mechanism of ineffective erythropoiesis in beta-thalassemia/Hb E disease. AB - BACKGROUND: Cells respond to stress stimuli through a number of response pathways, of which one of the most important and well characterized is the unfolded protein response. Despite a large body of work which suggests that stress in erythroblasts may play a pivotal role in the pathogenesis of beta thalassemia/Hb E disease, this pathway remains uninvestigated. DESIGN AND METHODS: Day 10 erythroblasts from normal controls and beta-thalassemia/Hb E patients were subjected to internal (treatment with tunicamycin) and external (serum and growth factor withdrawal) stress stimuli and the activation of the unfolded protein response pathway was investigated. RESULTS: Normal erythroblasts responded to both internal and external stress by activating the unfolded protein response (UPR) pathway while in contrast, erythroblasts from beta-thalassemia/Hb E patients only showed activation of the unfolded protein response pathway in response to internal stress. This was reflected by a markedly increased induction of apoptosis in serum and growth factor deprived beta-thalassemia/Hb E erythroblasts as compared to control cells. Modulation of the levels of intracellular Ca(2+) in thalassemic erythroblasts restored UPR activation during serum deprivation and significantly reduced the level of serum deprivation induced apoptosis to control levels. CONCLUSIONS: These results suggest the failure of thalassemic erythroblasts to cope with cellular stress caused by an impaired UPR function as a result of high Ca(2+) levels may exacerbate thalassemic cell death during erythropoiesis. PMID- 20015892 TI - Constitutional mismatch repair deficiency and childhood leukemia/lymphoma--report on a novel biallelic MSH6 mutation. AB - Biallelic mutations of mismatch repair genes cause constitutional mismatch repair deficiency associated with an increased risk for childhood leukemia/lymphoma. We report on a case with constitutional mismatch repair deficiency caused by a novel MSH6 mutation leading to a T-cell lymphoma and colonic adenocarcinoma at six and 13 years of age, respectively. A review of the literature on hematologic malignancies in constitutional mismatch repair deficiency showed that in almost half of the 47 known constitutional mismatch repair deficiency families, at least one individual is affected by a hematologic malignancy, predominantly T-cell lymphomas. However, diagnosing constitutional mismatch repair deficiency may be difficult when the first child is affected by leukemia/lymphoma, but identification of the causative germline mutation is of vital importance: (i) to identify relatives at risk and exclude an increased risk in non-mutation carriers; (ii) to prevent hematopoietic stem cell transplantation from sibling donors also carrying a biallelic germline mutation; and (iii) to implement effective surveillance programs for mutation carriers, that may reduce constitutional mismatch repair deficiency-associated mortality. PMID- 20015890 TI - Daily practice management of myelodysplastic syndromes in France: data from 907 patients in a one-week cross-sectional study by the Groupe Francophone des Myelodysplasies. AB - BACKGROUND: There is little published information on the everyday clinical management of myelodysplastic syndromes in real world practice. DESIGN AND METHODS: We conducted a cross-sectional study of all patients with myelodysplastic syndromes attending 74 French centers in a 1-week period for inpatient admission, day-hospital care or outpatient visits. RESULTS: Nine hundred and seven patients were included; 67.3% had lower-risk myelodysplastic syndromes (International Prognostic Scoring System: low or intermediate-1). Karyotype had been analyzed in 82.5% of the cases and was more often of intermediate or poor risk in patients under 65 years old compared with those who were older. Red blood cell transfusions accounted for as many as 31.4% of the admissions. Endogenous erythropoietin level was less than 500 IU/L in 88% of the patients tested. Erythroid stimulating agents had been or were being used in 36.8% of the lower risk patients, iron chelation in 31% of lower risk patients requiring red blood cell transfusions and lenalidomide in 41% of lower risk patients with del 5q. High-dose chemotherapy, hypomethylating agents, low dose cytarabine and allogeneic stem cell transplantation had been or were being used in 14.8%, 31.1%, 8.8% and 5.1%, respectively, of higher-risk patients. CONCLUSIONS: Karyotype is now assessed in most patients with myelodysplastic syndromes, and patients under 65 years old may have more aggressive disease. Apart from erythroid-stimulating agents and, in higher-risk myelodysplastic syndromes, hypomethylating agents, specific treatments are used in a minority of patients with myelodysplastic syndromes and red blood cell transfusions still represent the major reason for hospital admission. PMID- 20015893 TI - Molecular analysis of 42 patients with congenital dyserythropoietic anemia type II: new mutations in the SEC23B gene and a search for a genotype-phenotype relationship. AB - BACKGROUND: The most frequent form of congenital dyserythropoietic anemia is the type II form. Recently it was shown that the vast majority of patients with congenital dyserythropoietic anemia type II carry mutations in the SEC23B gene. Here we established the molecular basis of 42 cases of congenital dyserythropoietic anemia type II and attempted to define a genotype-phenotype relationship. DESIGN AND METHODS: SEC23B gene sequencing analysis was performed to assess the diversity and incidence of each mutation in 42 patients with congenital dyserythropoietic anemia type II (25 described exclusively in this work), from the Italian and the French Registries, and the relationship of these mutations with the clinical presentation. To this purpose, we divided the patients into two groups: (i) patients with two missense mutations and (ii) patients with one nonsense and one missense mutation. RESULTS: We found 22 mutations of uneven frequency, including seven novel mutations. Compound heterozygosity for a missense and a nonsense mutation tended to produce a more severe clinical presentation, a lower reticulocyte count, a higher serum ferritin level, and, in some cases, more pronounced transfusion needs, than homozygosity or compound heterozygosity for two missense mutations. Homozygosity or compound heterozygosity for two nonsense mutations was never found. CONCLUSIONS: This study allowed us to determine the most frequent mutations in patients with congenital dyserythropoietic anemia type II. Correlations between the mutations and various biological parameters suggested that the association of one missense mutation and one nonsense mutation was significantly more deleterious that the association of two missense mutations. However, there was an overlap between the two categories. PMID- 20015894 TI - Mitotic recombination and compound-heterozygous mutations are predominant NF1 inactivating mechanisms in children with juvenile myelomonocytic leukemia and neurofibromatosis type 1. AB - Children with neurofibromatosis type 1 (NF-1), being constitutionally deficient for one allele of the NF1 gene, are at greatly increased risk of juvenile myelomonocytic leukemia (JMML). NF1 is a negative regulator of RAS pathway activity, which has a central role in JMML. To further clarify the role of biallelic NF1 gene inactivation in the pathogenesis of JMML, we investigated the somatic NF1 lesion in 10 samples from children with JMML/NF-1. We report that two thirds of somatic events involved loss of heterozygosity (LOH) at the NF1 locus, predominantly caused by segmental uniparental disomy of large parts of chromosome arm 17q. One-third of leukemias showed compound-heterozygous NF1-inactivating mutations. A minority of cases exhibited somatic interstitial deletions. The findings reinforce the emerging role of somatic mitotic recombination as a leukemogenic mechanism. In addition, they support the concept that biallelic NF1 inactivation in hematopoietic progenitor cells is required for transformation to JMML in children with NF-1. PMID- 20015895 TI - "Silent virtuous teachers": anatomical dissection in Taiwan. AB - Steven C Lin, Julia Hsu, and Victoria Y Fan describe how the dissection course at Tzu Chi College of Medicine in Taiwan has motivated and inspired people to donate their bodies for scientific endeavours, an otherwise culturally avoided gesture in the East. PMID- 20015897 TI - A gold elixir of youth in the 16th century French court. PMID- 20015898 TI - Django's Hand. PMID- 20015904 TI - Wards of the roses. PMID- 20015903 TI - Topical intranasal corticosteroids in 4-11 year old children with persistent bilateral otitis media with effusion in primary care: double blind randomised placebo controlled trial. AB - OBJECTIVE: To determine the clinical effectiveness of topical intranasal corticosteroids in children with bilateral otitis media with effusion. DESIGN: Double blind randomised placebo controlled trial. SETTING: 76 Medical Research Council General Practice Research Framework practices throughout the United Kingdom, between 2004 and 2007. PARTICIPANTS: 217 children aged 4-11 years who had at least one practice recorded episode of otitis media or a related ear problem in the previous 12 months, and with bilateral otitis media with effusion confirmed by a research nurse using otoscopy plus micro-tympanometry (B/B or B/C2, modified Jerger types). INTERVENTION: Mometasone furoate 50 microg or placebo spray given once daily into each nostril for three months. MAIN OUTCOME MEASURES: Proportions of children cured of bilateral otitis media with effusion assessed with tympanometry (C1 or A type) at one month (primary end point), three months, and nine months; adverse events; three month diary symptoms. Results 41% (39/96) of the topical steroid group and 45% (44/98) of the placebo group were cured in one or both ears at one month (difference favouring placebo 4.3% (95% confidence interval -9.3% to 18.1%). Poisson regression was done with adjustment for four pre-specified covariates (clinical severity, P=0.003; atopy, P=0.67; age, P=0.92; season, P=0.71). The adjusted relative risk at one month was 0.97 (95% confidence interval 0.74 to 1.26). At three months, 58% of the topical steroid group and 52% of the placebo group were cured (relative risk 1.23, 0.84 to 1.80). Diary symptoms did not differ between the two groups, and no significant harms were reported. CONCLUSIONS: Topical steroids are unlikely to be an effective treatment for otitis media with effusion in general practice. High rates of natural resolution occurred by 1-3 months. TRIAL REGISTRATION: Current Controlled Trials ISRCTN38988331; National Research Register NO575123823; MREC 03/11/073. PMID- 20015906 TI - Where have all the hospital flowers gone? PMID- 20015907 TI - Lying obliquely--a clinical sign of cognitive impairment: cross sectional observational study. AB - OBJECTIVE: To determine if failure to spontaneously orient the body along the longitudinal axis of a hospital bed when asked to lie down is associated with cognitive impairment in older patients. DESIGN: Cross sectional observational study. SETTING: Neurology department of a university hospital in Germany. PARTICIPANTS: Convenience sample of 110 older (>or=60 years) inpatients with neurological conditions and 23 staff neurologists. MAIN OUTCOME MEASURES: The main outcome measure was the association between the angle of the body axis and the results of three cognitive screening tests (mini-mental state examination, DemTect, and clock drawing test). Staff doctors were shown photographs of a model taken at a natural viewing able to determine their subjective perspective of what constitutes oblique. RESULTS: 110 neurological inpatients (mean age 70.9 (SD 6.8) years) were included after exclusions. Evidence of cognitive impairment was found in 34, with scores indicating dementia in eight, according to the mini-mental state examination, and in 11 according to the DemTect. Across all patients, the mean angular deviation of the body axis from the longitudinal axis of the bed (range 0-23 degrees ) correlated linearly with the mini-mental state examination (r=-0.480), DemTect (r=-0.527), and the clock drawing test (r=-0.552) scores (P<0.001 for all), even after removing age as a covariate. Overall, 90% of staff neurologists considered a minimal body angle of 7 degrees to be oblique. Angular deviation of at least 7 degrees predicted cognitive impairment according to the three different tests, with specificities between 89% and 96% and sensitivities between 27% and 50%. CONCLUSION: Clinicians might suspect cognitive impairment in mobile older inpatients with neurological disorders who spontaneously position themselves obliquely when asked to lie on a bed. PMID- 20015908 TI - Stigma and prejudice in Tintin. PMID- 20015909 TI - The Spanish influenza pandemic seen through the BMJ's eyes: observations and unanswered questions. PMID- 20015910 TI - In praise of the physical examination. PMID- 20015912 TI - Pattern of oseltamivir ingestions reported to Texas poison centers. AB - During serious influenza outbreaks, the number of oseltamivir exposures reported to poison centers might be expected to increase. This investigation describes the pattern of oseltamivir ingestions reported to Texas poison centers during 2000 2008. Of 298 total ingestions, 91.9% occurred in December-March, 76.8% involved patients aged 0-19 years, 72.5% resulted from therapeutic error, 90.0% were managed on-site, and 80.0% had no effect. The most frequently reported adverse clinical effects were vomiting (7.5%), nausea (3.8%), and abdominal pain (3.8%). Oseltamivir ingestions were reported to Texas poison centers primarily during periods of influenza outbreak. Most involved children, resulted from therapeutic error, and were managed on-site without serious outcome. PMID- 20015913 TI - Why does the provision of home mechanical ventilation vary so widely? AB - There is wide variation in the provision of home mechanical ventilation (HMV) throughout Europe, but the provision of home mechanical ventilation can also vary within countries. In 2008, the overall prevalence of HMV in Norway was 19.9/100,000, and there were huge regional differences in treatment prevalence. The aim of this study is to find explanations for these differences. We gathered multidisciplinary respondents involved in HMV treatment from five hospitals in five different counties to six focus group conversations to explore respondents' views of their experiences systematically. We based the analysis on grounded theory. We found that uneven distribution of ''enthusiasm'' between hospitals seems to be an important factor in the geographical distribution of HMV. Furthermore, we found that the three subcategories, ''high competence,'' ''spreading competence,'' and ''multidisciplinary collaboration,'' are developed and used systematically in counties with ''enthusiasm.'' This culture is the main category, which might explain the differences, and is described as ''wise enthusiasm.'' The last subcategory is ''individual attitudes'' about HMV among decision-making physicians. The most important factor is most likely the uneven distribution of highly skilled enthusiasm between hospitals. Individual attitudes about HMV among the decision makers may also explain why the provision of HMV varies so widely. Data describing regional differences in the prevalence of HMV within countries is lacking. Further research is needed to identify these differences to ensure equality of provision of HMV. PMID- 20015914 TI - Impending cardiac tamponade as the initial presentation of thymoma in a systemic lupus erythematosus patient. PMID- 20015915 TI - Clinical associations with autoantibody reactivities to individual components of U1 small nuclear ribonucleoprotein. AB - The reactivities to individual U1 small nuclear ribonucleoprotein (snRNP) components and their relationship to clinical features in patients with anti-U1 snRNP antibodies were examined. We evaluated 114 patients with connective tissue disease whose sera were positive for anti-U1 snRNP antibodies, but negative for anti-Sm antibodies. Antibodies to the U1 snRNP polypeptides 70K, A, and C were detected using subunit-specific enzyme-linked immunosorbent assays and antibodies to U1 small nuclear RNA (snRNA) were identified by an immunoprecipitation assay using deproteinized HeLa cell extracts. The clinical features were retrospectively obtained by chart review and prospectively collected after study entry. The pattern of antibody reactivities to U1 snRNP components varied among patients. The frequency of anti-70K, anti-A, anti-C, and anti-U1 snRNA antibodies was 60%, 86%, 74%, and 46%, respectively. There was no relationship between each reactivity and the clinical findings, but the presence of reactivities to increasing numbers of U1 snRNP components was correlated with sclerodactyly, shortness of the sublingual frenulum, esophageal dysfunction, and a lack of persistent proteinurea (p < 0.05 for all comparisons). The detection of autoantibody reactivities to individual components of the U1 snRNP particle is potentially useful for predicting the clinical course in patients with connective tissue disease and anti-U1 snRNP antibodies. PMID- 20015916 TI - Cardiac autonomic profile in rheumatoid arthritis and systemic lupus erythematosus. AB - Neurological involvement is a well-documented issue in patients with systemic lupus erythematosus (SLE) and rheumatoid arthritis (RA). However, little is known about the involvement of the autonomic nervous system. This study was conducted to investigate autonomic nervous system dysfunction in patients with RA and SLE. Twenty-six RA patients, 38 SLE patients and 40 healthy controls were recruited from our in- and out-patient departments. Heart rate variability (HRV) parameters (the power of the high- [HF] and low-frequency [LF] band of haemodynamic time series, the ratio between low- and high-frequency components [LF/HF ratio], the power spectral density), baroreflex sensitivity (BRS) and beat-to-beat blood pressures were assessed by a novel non-invasive haemodynamic monitoring tool (Task Force Monitor [TFM], CNSystems Medizintechnik GmbH, Graz, Austria). Autonomic nervous system dysfunction was determined according to classical Ewing autonomic test battery. Furthermore, we implemented a secondary autonomic test score by modifying the Ewing test battery with additional criteria. Both the classical and modified Ewing test batteries have revealed that the frequencies of autonomic neuropathy were significantly higher in patient groups compared with controls (p < 0.001). Evaluation by TFM revealed that deterioration of sophisticated autonomic parameters (such as HRV and BRS) were more pronounced in the patient groups compared with controls. There was a significant association between BRS and Ewing test scores and abnormal BRS results were more frequent in patients with autonomic dysfunction according to Ewing test batteries. No relation was found between autonomic neuropathy and disease duration, disease activity and autoantibody positivity. Consequently, we believe that further large scale studies investigating cardiovascular autonomic neuropathy in rheumatic diseases should be carried out to verify our findings and manifest clinical consequences beyond these results. PMID- 20015917 TI - Exploring differences in referrals to a hospice at home service in two socio economically distinct areas of Manchester, UK. AB - In order to provide equitable access to hospice at home palliative care services, it is important to identify the socio-economic factors associated with poorer access. In this population-based study we aimed to test the inverse care law by exploring how socio-economic status and other key demographic indicators were associated with referral rates in two distinct areas (Salford and Trafford) served by the same service. Secondary data from the UK National Census 2001, North West Cancer Intelligence Service (2004) and hospice at home service referral data (2004-06) was collated for both areas. Descriptive analysis profiled electoral ward characteristics whilst simple correlations and regression modelling estimated associations with referral rates. Referral rates were lower and cancer mortality higher in the most deprived areas (Salford). Referral rates were significantly associated with deprivation, particularly multiple deprivation, but not significantly associated with cancer mortality (service model and resources available were held constant). At the population level, the socio-economic characteristics of those referred to hospice at home rather than service provision strongly predicted referral rates. This has implications for the allocation and targeting of resources and contributes important findings to future work exploring equitable access at organizational and professional levels. PMID- 20015918 TI - Gender imbalance in pediatric palliative care research samples. AB - We assessed the sampling performance of research on parental perspectives in pediatric palliative care and examined if and how gender imbalance was treated. We undertook a systematic review of parental perspectives research in pediatric palliative care using MEDLINE, CINAHL, and PsycINFO. Study selection inclusion criteria were: (1) published between 1988 and 2008; (2) in English; (3) conducted in North America; (4) focused on parents of children aged 0-18 years who were expected to die or had died; (5) had 'parent' in the title; and (6) focused on parents' experiences or on parents' perspectives regarding the child's illness/death. Keyword searches produced a list of 2103 studies, of which 45 met the criteria for inclusion. The ratio of mothers to fathers participating in the studies was examined. We found that there has been an increase in research on parental perspectives in pediatric palliative care over the last 5 years, but what constitutes 'parental' in this literature continues to be primarily 'maternal'. Mothers constituted 75% of the overall sample of parents. In only four studies was the gender imbalance addressed as one of the limitations of the study. There is a growing interest in parental perspectives in pediatric palliative care, but the research does not equally reflect the experiences and needs of mothers and fathers. Gender can shape experiences of both parenthood and grief; balanced gender sampling and accurate analysis is essential for research on 'parental perspectives'. Gender imbalance in research samples, designs, recruitment strategies, and data gathering methods must be addressed. PMID- 20015919 TI - How do nurses assess and manage breakthrough pain in specialist palliative care inpatient units? A multicentre study. AB - The aim of this qualitative study was to gain a better understanding of how nurses working on inpatient specialist palliative care units assess and manage breakthrough pain. Thematic analysis of semi-structured interviews with fifteen nurses from five different specialist palliative care units in the UK was undertaken. Themes identified have been broadly categorized into four main areas: defining breakthrough pain, assessing breakthrough pain, managing breakthrough pain, and attitudes/teamwork. Nurses had difficulty defining breakthrough pain as a distinct pain subtype and were often unable to differentiate it from poorly controlled background pain. This study highlights significant training needs and suggests that the theoretical work and recently published consensus recommendations around breakthrough pain now need to be translated into day-to day clinical practice. PMID- 20015920 TI - Symptom severity and distress in advanced cancer. AB - We determined the relationship between symptom severity and distress for multiple cancer symptoms, and examined patient demographic influences on severity and distress in advanced cancer. A Cochran-Armitage trend test determined whether symptom distress increased with severity. Chi-square, Fisher's exact test and logistic regression analysis examined moderate/severe ('clinically important') and distressful symptoms by age (65), gender, primary site group, and ECOG performance status. Forty-six symptoms were analyzed in 181 individuals. More than 50% of individuals with clinically important symptoms rated them as distressful. The median percentage of individuals with mild but still distressful symptoms was 25%, with a range of 0% (bad dreams) to 73% (sore mouth). In both univariate and multivariate analysis, younger ( 20 mm) was identified by 95% of patients. Reduced use of rescue medication (p < 0.001, SLF 400 microg) and improved global assessment of treatment (p = 0.0146, SLF 400 microg) confirmed these differences as clinically important. Nausea and dizziness were the most common treatment-related adverse effects. SLF appears to be a fast, effective and well-tolerated treatment for BTP. PMID- 20015923 TI - Benchmarking in-patient medication errors in specialist palliative care. PMID- 20015924 TI - Warm air and wicking fabric for apyrexial paraneoplastic sweats. PMID- 20015925 TI - Pharmacokinetics of vancomycin in adult cancer patients. AB - BACKGROUND AND OBJECTIVES: gram-positive infections are prevalent among cancer patients and vancomycin therapy is often initiated empirically. A typical vancomycin pharmacokinetics is observed in such patients. The aim of the study was to evaluate the pharmacokinetics of vancomycin in this patient population and compare it to that of normal population. METHOD AND RESULTS: the pharmacokinetics of vancomycin was examined retrospectively in two groups of patients - 18 cancer patients (age 43.4 +/- 22.1 years) and 13 patients without cancer (age 48.5 +/- 20.2 years). Following the administration of intermittent intravenous infusion of 15 mg/kg of vancomycin, peak and trough vancomycin serum concentration were determined after the third dose or at steady state as per standard of care. Vancomycin data were analyzed according to a one-compartment open model. Pharmacokinetic parameters such as clearance (CL), volume of distribution (Vd), and K elimination (ke) were calculated. Both Vd and CL were significantly higher in the cancer group (Mean Vd was 70 +/- 45 L in the cancer group and 31.1 +/- 8.3 L in the noncancer group, p-value 0.002; CL mean was 110.1 +/- 42 mL/min in the cancer group and 71.2 +/- 22.2 mL/min in the noncancer group, p-value 0.005). There was no significant difference in K elimination and half-life (t(1/2)). CONCLUSION: cancer patients may require higher than usual dosing regimens to ensure optimal therapeutic concentrations, since vancomycin CL and Vd is significantly higher in these patients, a dosing schedule as high as 60 mg/kg/day may be needed for cancer patients. PMID- 20015926 TI - Arsenic trioxide and olanzapine co-administration: case analysis. AB - QTc prolongation is associated with arsenic trioxide (ATO) treatment of acute promyelocytic leukemia (APL). Olanzapine was safely co-administered with ATO to treat co-morbid psychiatric diagnoses. It is important to closely monitor for drug-drug interactions and cumulative drug adverse effects in patients receiving oncology agents and psychotropics. Further research is indicated to determine risk/benefit profiles of psychotropics co-administered with ATO. In light of current limited data, co-administration of psychotropics with ATO should be reported presenting both instances wherein no interactions are noted and those with adverse effects. PMID- 20015927 TI - Chemotherapy dose intensity reductions due to adverse drug reactions in an oncology outpatient setting. AB - OBJECTIVE: the aim of this study was to establish the incidence of adverse drug reactions (ADRs) that prevent administration of planned dose intensity antineoplastic chemotherapy in clinical practice. METHODS: a retrospective cohort study was made of adult oncology patients who received intravenous chemotherapy in an outpatient ward during 2005. Dose delays, dose reductions, dose omissions, or treatment discontinuations with respect to the planned chemotherapy regimen were obtained, and causes were evaluated to identify ADRs. RESULTS: a total of 532 patients were included, involving the administration of 631 chemotherapy regimens. The total number of cycles evaluated was 3553. The incidence of nonadherence to planned treatment due to ADRs was 12.9% cycles (95% CI: 11.8 14.0). Differences according to patient's age were found, patients younger than 65 years showed higher rates of nonadherence due to ADRs. Myelosuppression was the main cause of dose delays, and gastrointestinal and neurological toxicity were reported as the main causes of dose reductions. CONCLUSIONS: the incidence of ADRs preventing planned dose intensity administration has been estimated. Follow-up of nonadherence to antineoplastic chemotherapy prescriptions can be used to evaluate tolerance in the clinical setting. Continuous evaluation of security in clinical practice provides information on benefits outweighing the deleterious effects in specific patient populations where the available data are limited. PMID- 20015928 TI - Atypical presentation of acute neurotoxicity secondary to oxaliplatin. AB - Neurotoxicity is the main and dose-limiting toxicity of oxaliplatin. It may produce two different syndromes, acute and chronic. We describe here a case of a patient with an acute syndrome with the particularity of affecting only contralateral hemibody to arm of infusion. A 62-year-old female diagnosed with stage IV colon cancer, underwent palliative treatment with combination of oxaliplatin (130 mg/m( 2) on day 1), capecitabine (1.250 mg/m(2) bid on days 1 to 14 every 3 weeks), and bevacizumab. Thirty minutes after cycle 1 oxaliplatin infusion, which was into the left arm, she experienced right hemibody paresthesia with muscle cramping of her right calf. She associated dysphonia and painful jamming sensation in her right upper limb with difficulty to release grip. She noted also undulating movements under the skin of her right lower extremity. She was unable to stand or walk. She was given intravenous magnesium sulfate and calcium gluconate and after 3 h all her symptoms were solved. Subsequent doses were reduced by 25% and the infusions were prolonged to 3 h and the patient tolerated well except minimal paresthesia in her right hand lasting few minutes. PMID- 20015929 TI - Review and revision of clinical practice of using G-CSF after autologous and allogeneic hematopoietic stem cell transplantation at UCSD. AB - There was no consensus on the optimal use of G-CSF after hematopoietic stem cell transplantation. In this study, the practice of using G-CSF, based on the CD34(+) cell number, at the University of California, San Diego Blood and Marrow Transplant Unit (UCSD BMT) was evaluated by performing a five-year retrospective analysis of data from patients undergoing autologous and allogeneic transplantation. Various outcomes, such as time to neutrophil and platelet engraftment and length of post-transplant hospital stay are assessed in relation to use of G-CSF and number of CD34(+) cells infused. It has been found that the use of G-CSF is associated with faster neutrophil engraftment and shorter length of post-transplant hospital stay without affecting time to platelet engraftment in patients undergoing autologous transplantation. In addition, the number of CD34(+) cells do not influence outcomes in autologous and allogeneic transplant patients if they are treated with G-CSF. As a result of this evaluation, the G CSF protocol at UCSD BMT Unit is revised. The main change is to implement the use of G-CSF in all patients undergoing autologous transplantation regardless of the number of CD34( +) cells. No changes in the allogeneic transplantation protocol are made as a result of this analysis. PMID- 20015930 TI - Fatal hepatic and renal toxicity as a complication of trabectedin therapy for radiation-induced sarcoma. AB - Trabectedin therapy was prescribed for a patient with radiation-induced sarcoma. Two doses of trabectedin were given before therapy was discontinued with the patient experiencing renal and liver failure. Despite discontinuing trabectedin the patient continued to experience increases in liver transaminases, bilirubin, blood urea nitrogen, and serum creatinine. Hemodialysis was initiated with no improvement. With all other causes being ruled out, trabectedin likely caused hepatic and renal failure leading to death in this patient. Recent literature suggests that patients may benefit from prophylactic dexamethasone as a means of reducing hepatic toxicity. PMID- 20015931 TI - Mutations of p53 gene in breast cancer in the Egyptian province of Dakahliya. AB - BACKGROUND: Breast cancer is the most common malignancy among females worldwide. Molecular analysis of p53 is likely to have value in diagnosis, prognosis, and treatment of breast cancer. OBJECTIVE: To study the frequency and spectrum of p53 gene mutations in breast cancer patients residing Al Dakahliya district in the north of Egypt. MATERIALS AND METHODS: Thirty patients with cancer breast as well as 10 controls were evaluated for p53 status by flow-cytometry, PCR-SSCP, and sequencing analysis. RESULTS: P53 mutations were evident in five breast cancer patients (17%) including two missense mutations (A218 T and R279 G) in exon 6, 8; nonsense mutations (S297stop and Y159stop) in exon 8, 5, respectively, and frame shift mutation (M133 fs) in exon 5. p53 mutations were associated with invasive ductal carcinoma, large tumor size, and advanced disease stage CONCLUSION: p53 gene mutations is potentially responsible for pathogenesis and clinical aggressiveness of breast cancer in our locality. PMID- 20015932 TI - Current challenges in European oncology pharmacy practice. AB - BACKGROUND: The demand for pharmacy cancer services is expected to at least double over the next 10 years, as the population ages and new treatments are introduced. Safe and efficient handling of cytotoxic products minimises risks to staff and reduces medication errors. OBJECTIVES: To identify and describe strategies for coping safely and effectively with heavier workloads in the hospital oncology pharmacy, currently and in the future. METHODS: The PubMed database was searched for literature on approaches to safe handling of antineoplastic agents and to decreasing medication errors in the hospital pharmacy. Articles that were judged to be of prime importance to the hospital oncologist were reviewed. These safety concepts are put into the context of contemporary hospital oncology pharmacy practice through discussion of key issues, including increased demand, the role of the pharmacist in determining the hospital formulary, and growth in patient preferences for oral chemotherapy. Recommendations on best practices are also provided, based on relevant literature and author experience. CONCLUSIONS: Efficient, safe hospital pharmacy operations can be aided by capacity planning, dose banding, and knowledge of novel products and procedures that can reduce risks to health while increasing the number of patients who are safely treated. Consideration may also be given to the economic role of oncology pharmacists in formulary development. PMID- 20015933 TI - Successful administration of a low dose of calcium nadroparin in patients suffering from pulmonary embolism and brain metastases: a report of two cases. AB - OBJECTIVE: To focus on the optimal management of thromboembolic complication in patients who have undergone chemotherapy with concomitant brain metastases and referred to a Division of Clinical Oncology. BACKGROUND: Thromboembolic diseases are common events in cancer patients due to clotting activation by tumor cells. On the other hand, brain metastases are common complication of systemic cancers. Postmortem studies show that a quarter of patients dying from cancer have intracranial metastases. Brain metastases and pulmonary embolism are life threatening conditions because of the risk of fatal endocranic hypertension and severe dyspnea. Calcium nadroparin is a low molecular weight heparin usually administered in patient with venous thromboembolism at a dose level of 180 IU/kg/daily. CASE SUMMARIES: The authors report the cases of two patients with intracranial metastases and pulmonary embolism-related dyspnea successfully treated with low dose of calcium nadroparin. A patient suffering from metastatic breast cancer and another one with metastatic nonsmall cell lung cancer were recently referred to our department because of severe dyspnea occurring during chemotherapy treatment. Both patients had cerebellar intracranial metastases. Massive pulmonary embolisms were shown by means of the computerized tomography. Despite the administration of a lower heparin dose than the usual one, around three-quarters of the calcium nadroparin daily conventional dose, quickly regressed dyspnea. Significant pulmonary embolism regression was revealed with computerized tomography scan within 8 weeks from the beginning of the thromboembolic complications. None of the patients showed any heparin treatment related complications. CONCLUSION: The authors conclude that, with regard to cancer patients carrying brain metastases who require anti-coagulant therapy, increased risk of intracranial hemorrhage should be kept in mind. An initial low molecular weight heparin dose reduction could be effective, and safely administered, also in case of pulmonary embolism with severe dyspnea. PMID- 20015934 TI - Colour, turbidity and COD removal from old landfill leachate by coagulation flocculation treatment. AB - The application of different coagulants and flocculants to leachate from an old landfill to determine the optimum conditions for removal of organic matter, colour and turbidity is studied. Ferric chloride, aluminium sulphate, aluminium polychloride (PAX) and polyacrylamide polyelectrolytes were tested. Higher pollutant removals (73% COD, 98% colour and 100% turbidity) were obtained using ferric chloride at pH 5.0-5.5 and for a dosage of 0.6 g Fe l(- 1). The volume of sludge generated after centrifugation represents about 4.0-4.6% when ferric chloride or aluminium sulphate is used, and 10% when employing aluminium polychloride. When flocculants were also added, the results obtained were similar to those found when adding only coagulants, although a considerable increase in the settling rate was obtained. PMID- 20015936 TI - On-site production of crude glucoamylase for kitchen waste hydrolysis. AB - Kitchen waste from dining rooms accounts for a considerable proportion of municipal solid garbage, and economical recycle ways are needed to be developed. This study investigated glucoamylase production from kitchen waste and the feasibility of kitchen waste hydrolysis by the crude enzymes produced. The key problems of high water content and poor porosity in kitchen waste for glucoamylase production under solid-state fermentation could be solved readily by the addition of corn stover or paddy husk. As a support medium, corn stover was better than paddy husk. Smashed kitchen waste (sKW) mixed with corn stover in the ratio of 3.75 : 1 (dry basis) produced 1838 U g(-1) of glucoamylase by Aspergillus niger UV-60 within 96 h. The enzyme productivity from kitchen waste was over two-fold higher than that from wheat bran with additional nutrients. Without any recovery treatment, the produced glucoamylase could be used directly to hydrolyse sKW slurry. The optimum enzyme dose 8% (crude enzyme/kichen waste, w/w) was not too big, and was sufficient to hydrolyse 10% (dry basis) sKW slurry to produce a maximum amount of reducing sugar of 55.4 g L(-1). PMID- 20015937 TI - Vermicomposting of olive oil mill wastewaters. AB - The disposal of olive oil mill wastewaters (OMW) represents a substantial environmental problem in Italy. A vermicompost process could be an alternative and valid method for the management of OMW. In a laboratory experiment, the OMW were absorbed onto a ligno-cellulosic solid matrix and 30 adult earthworms of Eisenia fetida specie were added. The experiment was carried out for 13 weeks. The number of earthworms increased throughout the experimental period and after 2 weeks about 90% of the earthworms had become sexually mature. The decrease in total organic carbon (about 35%), C : N ratio (from 31.2 to 12.3) and biochemical parameters (hydrolytic enzymes averagely 40% and dehydrogenase 23%), and the increase in humification rate (pyrophosphate extractable carbon (PEC) from 17.6 to 33.3 mg g(-1), and PEC : water-soluble carbon from 1.76 to 2.97) indicated the mineralization and the stabilization of organic matter at the end of the vermicomposting process. At the end of the experiment, the extracellular beta glucosidase, phosphatase, urease and protease activities, measured in the pyrophosphate extract of the vermicompost, were found to be always higher or equal to that measured at the beginning of the vermicomposting process, suggesting that the enzymes bound to humic matter resisted biological attack and environmental stress. Moreover, the results obtained from the phyto-test showed that the OMW lose their toxicity and stimulate plant germination and growth. PMID- 20015938 TI - Evaluation of biological landfill leachate treatment incorporating struvite precipitation and powdered activated carbon addition. AB - Direct application of aerobic biodegradation for leachate treatment is not feasible due to high concentrations of nitrogen and chemical oxygen demand. Several potential leachate treatment schemes incorporating struvite precipitation as pretreatment and two types of activated sludge processes (conventional activated sludge and batch decant reactor (intermittent cycle extended aeration system [ICEAS process]) with and without addition of powdered activated carbon (PAC) were evaluated in this study. The hydraulic retention times (HRT) of 6 h and 12 h were applied in the biological stages of each process. Treatment schemes incorporating PAC addition showed acceptable results. In the conventional activated sludge process, total chemical oxygen demand (TCOD), soluble chemical oxygen demand (SCOD), NH(3), and P removal rates were 87, 84, 98.3, and 94%, respectively, with HRT of 6 h and 95.8, 95.1, 99.1, and 98.7% with HRT of 12 h. For the ICEAS process, removal rates were 89.3, 87.9, 98.2, and 94% with HRT of 6 h and 95.8, 95, 99.2, and 98.7% with HRT of 12 h. On the basis of these results, it can be concluded that struvite precipitation, followed by activated sludge process with PAC addition, can be a strong alternative leachate treatment, achieving the standards set for effluent discharges to receiving waters. PMID- 20015939 TI - Extracellular glutathione peroxidase (Gpx3) binds specifically to basement membranes of mouse renal cortex tubule cells. AB - Glutathione peroxidase-3 (Gpx3), also known as plasma or extracellular glutathione peroxidase, is a selenoprotein secreted primarily by kidney proximal convoluted tubule cells. In this study Gpx3(-/-) mice have been produced and immunocytochemical techniques have been developed to investigate Gpx3 metabolism. Gpx3(-/-) mice maintained the same whole-body content and urinary excretion of selenium as did Gpx3(+/+) mice. They tolerated selenium deficiency without observable ill effects. The simultaneous knockout of Gpx3 and selenoprotein P revealed that these two selenoproteins account for >97% of plasma selenium. Immunocytochemistry experiments demonstrated that Gpx3 binds selectively, both in vivo and in vitro, to basement membranes of renal cortical proximal and distal convoluted tubules. Based on calculations using selenium content, the kidney pool of Gpx3 is over twice as large as the plasma pool. These data indicate that Gpx3 does not serve in the regulation of selenium metabolism. The specific binding of a large pool of Gpx3 to basement membranes in the kidney cortex strongly suggests a need for glutathione peroxidase activity in the cortical peritubular space. PMID- 20015940 TI - Functional characterization of transient receptor potential channels in mouse urothelial cells. AB - The bladder urothelium is currently believed to be a sensory structure, contributing to mechano- and chemosensation in the bladder. Transient receptor potential (TRP) cation channels act as polymodal sensors and may underlie some of the receptive properties of urothelial cells. However, the exact TRP channel expression profile of urothelial cells is unclear. In this study, we have performed a systematic analysis of the molecular and functional expression of various TRP channels in mouse urothelium. Urothelial cells from control and trpv4 /- mice were isolated, cultured (12-48 h), and used for quantitative real-time PCR, immunocytochemistry, calcium imaging, and whole cell patch-clamp experiments. At the mRNA level, TRPV4, TRPV2, and TRPM7 were the most abundantly expressed TRP genes. Immunohistochemistry showed a clear expression of TRPV4 in the plasma membrane, whereas TRPV2 was more prominent in the cytoplasm. TRPM7 was detected in the plasma membrane as well as cytoplasmic vesicles. Calcium imaging and patch-clamp experiments using TRP channel agonists and antagonists provided evidence for the functional expression of TRPV4, TRPV2, and TRPM7 but not of TRPA1, TRPV1, and TRPM8. In conclusion, we have demonstrated functional expression of TRPV4, TRPV2, and TRPM7 in mouse urothelial cells. These channels may contribute to the (mechano)sensory function of the urothelial layer and represent potential targets for the treatment of bladder dysfunction. PMID- 20015941 TI - Disparate effects on renal and oxidative parameters following RAGE deletion, AGE accumulation inhibition, or dietary AGE control in experimental diabetic nephropathy. AB - Advanced glycation end products (AGEs) and the receptor for AGEs (RAGE) generate ROS, and therefore this study evaluated the effects of RAGE deletion, decreasing AGE accumulation, or lowering dietary AGE content on oxidative parameters in diabetic nephropathy (DN). Control and diabetic male wild-type and RAGE-deficient (RAGE-/-) mice were fed high- or low-AGE diets, with two groups given the inhibitor of AGE accumulation, alagebrium chloride, and followed for 24 wk. Diabetic RAGE-/- mice were protected against albuminuria, hyperfiltration, glomerulosclerosis, decreased renal mitochondrial ATP production, and excess generation of both mitochondrial and cytosolic superoxide. Whereas glomerulosclerosis, tubulointerstitial expansion, and hyperfiltration were improved in diabetic mice treated with alagebrium, there was no effect on urinary albumin excretion. Both diabetic RAGE-/- and alagebrium-treated mice had an attenuation of renal RAGE expression and decreased renal and urinary AGE (carboxymethyllysine) levels. Low-AGE diets did not confer renoprotection, lower the AGE burden or renal RAGE expression, or improve cytosolic or mitochondrial superoxide generation. Renal uncoupling protein-2 gene expression and mitochondrial membrane potential were attenuated by all therapeutic interventions in diabetic mice. In the present study, diverse approaches to block the AGE-RAGE axis had disparate effects on DN, which has potential clinical implications for the way this axis should be targeted in humans. PMID- 20015942 TI - Troglitazone ameliorates high glucose-induced EMT and dysfunction of SGLTs through PI3K/Akt, GSK-3beta, Snail1, and beta-catenin in renal proximal tubule cells. AB - Peroxisome proliferator-activated receptor-gamma (PPARgamma) agonists ameliorate renal fibrotic lesions in diabetic nephropathy. However, the effects of the agonists on the epithelial-mesenchymal transition (EMT) linked to membrane transport dysfunction are unknown. The present study aimed to verify the effects of the PPARgamma agonist troglitazone on high glucose (HG)-induced EMT in primary cultured renal proximal tubular epithelial cells (PTCs). HG (25 mM) as well as hydrogen peroxide (H(2)O(2)) and transforming growth factor-beta1 (TGF-beta1) decreased expression of epithelial cell marker E-cadherin and increased the expression of the mesenchymal markers vimentin and alpha-smooth muscle actin (alpha-SMA). HG, H(2)O(2), and TGF-beta1 decreased Na(+)/H(+) exchangers (NHEs) or Na(+)-glucose cotransporters (SGLTs) and glucose uptake, showing membrane transport dysfunction. HG stimulated the production of cellular reactive oxygen species (ROS), and antioxidants blocked the HG-induced increase in phosphatidylinositol 3-kinase (PI3K)/Akt activation. Antioxidants and inhibitors of PI3K/Akt reversed HG-induced EMT protein expression. Inhibition of PI3K/Akt also blocked HG-induced glycogen synthase kinase-3beta (GSK-3beta) phosphorylation. HG and lithium chloride (GSK-3beta inhibitor) blocked Snail1 and beta-catenin activation. Moreover, transfection with Snail1 or beta-catenin small interfering RNA (siRNA) reversed HG-induced EMT protein expression. Importantly, HG decreased PPARgamma activation and troglitazone reversed HG-induced expression of PI3K/Akt, GSK-3beta, Snail1, and beta-catenin as well as EMT proteins. Finally, inhibitors of PI3K/Akt, Snail1/beta-catenin siRNA, and troglitazone blocking the HG-induced EMT restored glucose uptake in PTCs. In conclusion, HG induces EMT through ROS, PI3K/Akt, GSK-3beta, Snail, and beta-catenin. Subsequently, HG-induced EMT may result in SGLT dysfunction that is restored by the PPARgamma agonist troglitazone in primary cultured PTCs. PMID- 20015944 TI - Soluble Flt-1 gene therapy ameliorates albuminuria but accelerates tubulointerstitial injury in diabetic mice. AB - VEGF is recognized as a major mediator in the development of diabetic nephropathy. Soluble Flt-1 (sFlt-1) is the endogenous inhibitor of VEGF, and recently genetic overexpression of sFlt-1 in the podocyte was shown to be protective in murine diabetic nephropathy. In this study, we performed a translational study to determine whether an intramuscular gene transfer of sFlt-1 can prevent the progression of renal disease in diabetic db/db mice. Adeno associated virus-1 (AAV1) encoding human sFlt-1 in two different doses was intramuscularly administrated in db/db and wild-type mice. The sFlt-1-AAV1 treatment significantly increased serum sFlt-1 level at 4 and 8 wk. A dose that was developed in this study caused minimal abnormalities in normal mice but reduced albuminuria in diabetic db/db mice. In renal histology, sFlt-1 treatment at this dose had minimal effects on mesangial expansion in diabetic mice, whereas podocyte injury was significantly improved, at 8 wk. Unfortunately, tubulointerstitial injury was markedly exacerbated by sFlt-1 treatment in association with a reduction in endogenous VEGF expression and peritubular capillary loss. In conclusion, gene therapy with sFlt-1-AAV1 protects podocytes but accelerates tubulointerstitial injury in diabetic db/db mice. These data suggest systemic overexpression of sFlt-1 will not likely be useful for treating diabetic nephropathy. PMID- 20015943 TI - HIVAN phenotype: consequence of epithelial mesenchymal transdifferentiation. AB - Human immunodeficiency virus (HIV)-1-associated nephropathy (HIVAN) is characterized by proliferation of glomerular and tubular epithelial cells. We studied the role of epithelial mesenchymal transdifferentiation (EMT) in the development of HIVAN phenotype. Renal cortical sections from six FVB/N (control) and six Tg26 (HIVAN) mice were immunolabeled for PCNA, alpha-smooth muscle actin (alpha-SMA), fibroblast-specific protein-1 (FSP1), CD3, and F4/80. Since periglomerular cells (PGCs) and peritubular cells (PTCs) did not show any labeling for CD3 and F4/80 but showed labeling for alpha-SMA or FSP1, it appears that these were myofibroblasts that migrated from either glomerular or tubular sites, respectively. Occurrence of EMT was also supported by diminished expression of E-cadherin by renal epithelial cells in Tg26 mice. Interestingly, Tg26 mice also showed enhanced renal tissue expression of ZEB2; henceforth, it appears that transcription of molecules required for maintenance of de novo renal epithelial cell phenotype was suppressed. To evaluate the role of ANG II, Tg26 mice in groups of three were administered either normal saline or telmisartan (an AT1 receptor blocker) for 2 wk, followed by evaluation for renal cell EMT. Renal cortical section of Tg26 mice showed a sevenfold increase (P < 0.001) in parietal epithelial cell (PEC)-PGC and a threefold increase (P < 0.01) in tubular cell (TC)-PTC proliferation (PCNA-positive cells). Similarly, both PECs-PGCs and TCs PTCs in Tg26 mice showed enhanced expression of alpha-SMA and FSP1. Both PECs and podocytes contributed to the glomerular proliferative phenotype, but the contribution of PECs was much greater. Telmisartan-receiving Tg26 mice (TRM) showed attenuated number of proliferating PECs-PGCs and TCs-PTCs compared with saline-receiving Tg26 mice (SRM). Similarly, TRM showed diminished expression of alpha-SMA and FSP1 by both PECs-PGCs and TCs-PTCs compared with SRM. We conclude that EMT contributes to the manifestation of the proliferative phenotype in HIVAN mice. PMID- 20015945 TI - Connective tissue growth factor (CTGF/CCN2) is increased in peritoneal dialysis patients with high peritoneal solute transport rate. AB - Peritoneal fibrosis (PF) is an important complication of peritoneal dialysis (PD) therapy that often occurs in association with peritoneal high transport rate and ultrafiltration failure (UFF). To study the possible pathogenic role of connective tissue growth factor (CTGF) in the relationship of PF and UFF, dialysate CTGF contents (n = 178) and tissue CTGF expression (n = 61) were investigated by ELISA, real-time PCR, immunohistochemistry, and in situ hybridization. CTGF production with and without TGF-beta1 stimulation in human peritoneal mesothelial cells (HPMC) from the spent patients' peritoneal dialysate (n = 32) was studied in vitro. The dialysate-to-plasma ratio for creatinine (D/P Cr) was positively correlated to dialysate CTGF concentration and estimated local peritoneal production of CTGF. CTGF mRNA expression was 11.4-fold higher in peritoneal membranes with UFF than in pre-PD renal failure peritoneum and was correlated with thickness of the peritoneum. CTGF protein and mRNA were detected in mesothelium and in fibroblast-like cells. In cultured HPMC, TGF-beta(1) induced expression of CTGF mRNA was increased at 12 and 24 h and was correlated with D/P Cr. In contrast, bone morphogenic protein-4 mRNA expression was inversely correlated with D/P Cr. Our results suggest that high peritoneal transport state is associated with fibrosis and increased peritoneal CTGF expression and production by mesothelial cells, which can be stimulated by TGF beta1. Dialysate CTGF concentration could be a biomarker for both peritoneal fibrosis and membrane function. Functional alteration of mesothelial cells may be involved in progression of peritoneal fibrosis in high transport state. PMID- 20015946 TI - NHE3 function and phosphorylation are regulated by a calyculin A-sensitive phosphatase. AB - Na+/H+ exchanger 3 (NHE3) is phosphorylated and regulated by multiple kinases, including PKA, SGK1, and CK2; however, the role of phosphatases in the dephosphorylation and regulation of NHE3 remains unknown. The purpose of this study was to determine whether serine/threonine phosphatases alter NHE3 activity and phosphorylation and, if so, at which sites. To this end, we first examined the effects of calyculin A [a combined protein phosphatase 1 (PP1) and PP2A inhibitor] and okadaic acid (a PP2A inhibitor) on general and site-specific NHE3 phosphorylation. Calyculin A induced a phosphorylation-dependent NHE3 gel mobility shift and increased NHE3 phosphorylation at serines 552 and 605. No change in NHE3 phosphorylation was detected after okadaic acid treatment. An NHE3 gel mobility shift was also evident in calyculin A-treated COS-7 cells transfected with either wild-type or mutant (S552A, S605G, S661A, S716A) rat NHE3. Since the NHE3 gel mobility shift occurred despite mutation of known phosphorylation sites, novel sites of phosphorylation must also exist. Next, we assayed NHE3 activity in response to calyculin A and okadaic acid and found that calyculin A induced a 24% inhibition of NHE3 activity, whereas okadaic acid had no effect. When all known NHE3 phosphorylation sites were mutated, calyculin A induced a stimulation of NHE3 activity, demonstrating a functional significance for the novel phosphorylation sites. Finally, we established that the PP1 catalytic subunit can directly dephosphorylate immunopurified NHE3 in vitro. In conclusion, our data demonstrate that a calyculin A-sensitive phosphatase, most likely PP1, is involved in the regulation and dephosphorylation of NHE3 at known and novel sites. PMID- 20015947 TI - Mixture-model based estimation of gene expression variance from public database improves identification of differentially expressed genes in small sized microarray data. AB - MOTIVATION: The small number of samples in many microarray experiments is a challenge for the correct identification of differentially expressed gens (DEGs) by conventional statistical means. Information from public microarray databases can help more efficient identification of DEGs. To model various experimental conditions of a public microarray database, we applied Gaussian mixture model and extracted bi- or tri-modal distributions of gene expression. Prior variance of Baldi's Bayesian framework was estimate for the analysis of the small sample sized datasets. RESULTS: First, we estimated the prior variance of a gene expression by pooling variances obtained from mixture modeling of large samples in the public microarray database. Then, using the prior variance, we identified DEGs in small sample-sized test datasets using the Baldi's framework. For benchmark study, we generated test datasets having several samples from relatively large datasets. Our proposed method outperformed other benchmark methods in terms of detecting gold-standard DEGs from the test datasets. The results may be a challenging evidence for usage of public microarray databases in microarray data analysis. PMID- 20015948 TI - r2cat: synteny plots and comparative assembly. AB - SUMMARY: Recent parallel pyrosequencing methods and the increasing number of finished genomes encourage the sequencing and investigation of closely related strains. Although the sequencing itself becomes easier and cheaper with each machine generation, the finishing of the genomes remains difficult. Instead of the desired whole genomic sequence, a set of contigs is the result of the assembly. In this applications note, we present the tool r2cat (related reference contig arrangement tool) that helps in the task of comparative assembly and also provides an interactive visualization for synteny inspection. PMID- 20015949 TI - RNAsnoop: efficient target prediction for H/ACA snoRNAs. AB - MOTIVATION: Small nucleolar RNAs are an abundant class of non-coding RNAs that guide chemical modifications of rRNAs, snRNAs and some mRNAs. In the case of many 'orphan' snoRNAs, the targeted nucleotides remain unknown, however. The box H/ACA subclass determines uridine residues that are to be converted into pseudouridines via specific complementary binding in a well-defined secondary structure configuration that is outside the scope of common RNA (co-)folding algorithms. RESULTS: RNAsnoop implements a dynamic programming algorithm that computes thermodynamically optimal H/ACA-RNA interactions in an efficient scanning variant. Complemented by an support vector machine (SVM)-based machine learning approach to distinguish true binding sites from spurious solutions and a system to evaluate comparative information, it presents an efficient and reliable tool for the prediction of H/ACA snoRNA target sites. We apply RNAsnoop to identify the snoRNAs that are responsible for several of the remaining 'orphan' pseudouridine modifications in human rRNAs, and we assign a target to one of the five orphan H/ACA snoRNAs in Drosophila. AVAILABILITY: The C source code of RNAsnoop is freely available at http://www.tbi.univie.ac.at/ -htafer/RNAsnoop PMID- 20015950 TI - Human papillomavirus-associated oropharyngeal cancer: a new clinical entity. AB - The incidence of oropharyngeal cancers is rising worldwide in both nonsmokers and nondrinkers. Epidemiology studies suggest a strong association between human papillomavirus (HPV) 16 infection, changing sexual behavior and cancer development. Despite initial presentation with locally advanced disease and poorly differentiated histology, HPV-associated oropharyngeal carcinoma is associated with a good prognosis because its response to chemotherapy and radiation. Clinicians should be aware of the risk of oropharyngeal cancer in young people to avoid unnecessary delay in diagnosis and treatment. A history of oral sex should be elicited in young patients with enlarged neck nodes and/or tonsillar masses. PMID- 20015951 TI - Healthcare and medical graduates of 2009: their reactions to four key proposals in the Scottish Government's strategy for tackling alcohol misuse. AB - AIMS: This study compares the views of final year medical, and nursing and allied health professional (NAHP) students in relation to four governmental proposals impacting on the sale and purchase of alcohol. METHODS: Against a background of political will to address alcohol abuse in Scotland and moves within the National Health Service promoting a shifting of professional roles, self-completed questionnaires were administered in spring 2009 through course websites and lectures to final year medical and NAHP students. RESULTS: Questionnaires were returned by 406 NAHPs and 121 medical students. Over three quarters of all students agreed with the proposed change to reduce the drink driving limit to 50 mg/100 ml blood. Less support was evident for the raising of the minimum legal purchase age for off-sales (37%), the banning of below cost price promotions of alcohol (47%) and minimum retail pricing (37%). However, there were differences between the NAHP and medical students in the case of the final two proposals; over 60% of the medical students agreed they would have a positive impact. For NAHPs, figures were 41% and 31%, respectively. CONCLUSIONS: Support for four key proposals outlined by the Scottish Government to address alcohol misuse varied. Only the suggestion to lower the drink driving limit received backing overall and within students in these professions. Effectiveness of proposed restrictions on the price of alcohol was less well regarded except by medical students. Evident gaps in knowledge around health guidelines, and the finding that almost half of NAHPs disagreed that they had the appropriate knowledge to advise patients about responsible drinking advice and alcohol misuse problems, suggest a need for improved undergraduate education and continued professional development with respect to public health aspects of alcohol use. PMID- 20015952 TI - European lactase persistence genotype shows evidence of association with increase in body mass index. AB - The global prevalence of obesity has increased significantly in recent decades, mainly due to excess calorie intake and increasingly sedentary lifestyle. Here, we test the association between obesity measured by body mass index (BMI) and one of the best-known genetic variants showing strong selective pressure: the functional variant in the cis-regulatory element of the lactase gene. We tested this variant since it is presumed to provide nutritional advantage in specific physical and cultural environments. We genetically defined lactase persistence (LP) in 31 720 individuals from eight European population-based studies and one family study by genotyping or imputing the European LP variant (rs4988235). We performed a meta-analysis by pooling the beta-coefficient estimates of the relationship between rs4988235 and BMI from the nine studies and found that the carriers of the allele responsible for LP among Europeans showed higher BMI (P = 7.9 x 10(-5)). Since this locus has been shown to be prone to population stratification, we paid special attention to reveal any population substructure which might be responsible for the association signal. The best evidence of exclusion of stratification came from the Dutch family sample which is robust for stratification. In this study, we highlight issues in model selection in the genome-wide association studies and problems in imputation of these special genomic regions. PMID- 20015954 TI - Comparative gene expression analysis of avian embryonic facial structures reveals new candidates for human craniofacial disorders. AB - Mammals and birds have common embryological facial structures, and appear to employ the same molecular genetic developmental toolkit. We utilized natural variation found in bird beaks to investigate what genes drive vertebrate facial morphogenesis. We employed cross-species microarrays to describe the molecular genetic signatures, developmental signaling pathways and the spectrum of transcription factor (TF) gene expression changes that differ between cranial neural crest cells in the developing beaks of ducks, quails and chickens. Surprisingly, we observed that the neural crest cells established a species specific TF gene expression profile that predates morphological differences between the species. A total of 232 genes were differentially expressed between the three species. Twenty-two of these genes, including Fgfr2, Jagged2, Msx2, Satb2 and Tgfb3, have been previously implicated in a variety of mammalian craniofacial defects. Seventy-two of the differentially expressed genes overlap with un-cloned loci for human craniofacial disorders, suggesting that our data will provide a valuable candidate gene resource for human craniofacial genetics. The most dramatic changes between species were in the Wnt signaling pathway, including a 20-fold up-regulation of Dkk2, Fzd1 and Wnt1 in the duck compared with the other two species. We functionally validated these changes by demonstrating that spatial domains of Wnt activity differ in avian beaks, and that Wnt signals regulate Bmp pathway activity and promote regional growth in facial prominences. This study is the first of its kind, extending on previous work in Darwin's finches and provides the first large-scale insights into cross species facial morphogenesis. PMID- 20015953 TI - Genetic modifiers of abnormal organelle biogenesis in a Drosophila model of BLOC 1 deficiency. AB - Biogenesis of lysosome-related organelles complex 1 (BLOC-1) is a protein complex formed by the products of eight distinct genes. Loss-of-function mutations in two of these genes, DTNBP1 and BLOC1S3, cause Hermansky-Pudlak syndrome, a human disorder characterized by defective biogenesis of lysosome-related organelles. In addition, haplotype variants within the same two genes have been postulated to increase the risk of developing schizophrenia. However, the molecular function of BLOC-1 remains unknown. Here, we have generated a fly model of BLOC-1 deficiency. Mutant flies lacking the conserved Blos1 subunit displayed eye pigmentation defects due to abnormal pigment granules, which are lysosome-related organelles, as well as abnormal glutamatergic transmission and behavior. Epistatic analyses revealed that BLOC-1 function in pigment granule biogenesis requires the activities of BLOC-2 and a putative Rab guanine-nucleotide-exchange factor named Claret. The eye pigmentation phenotype was modified by misexpression of proteins involved in intracellular protein trafficking; in particular, the phenotype was partially ameliorated by Rab11 and strongly enhanced by the clathrin-disassembly factor, Auxilin. These observations validate Drosophila melanogaster as a powerful model for the study of BLOC-1 function and its interactions with modifier genes. PMID- 20015955 TI - Interaction between Sdo1p and Btn1p in the Saccharomyces cerevisiae model for Batten disease. AB - Juvenile Batten disease is an autosomal recessive pediatric neurodegenerative disorder caused by mutations in the CLN3 gene. The CLN3 protein primarily resides in the lysosomal membrane, but its function is unknown. We demonstrate that CLN3 interacts with SBDS, the protein mutated in Shwachman-Bodian-Diamond syndrome patients. We demonstrate that this protein-protein interaction is conserved between Btn1p and Sdo1p, the respective yeast Saccharomyces cerevisiae orthologs of CLN3 and SBDS. It was previously shown that deletion of BTN1 results in alterations in vacuolar pH and vacuolar (H(+))-ATPase (V-ATPase)-dependent H(+) transport and ATP hydrolysis. Here, we report that an SDO1 deletion strain has decreased vacuolar pH and V-ATPase-dependent H(+) transport and ATP hydrolysis. These alterations result from decreased V-ATPase subunit expression. Overexpression of BTN1 or the presence of ionophore carbonyl cyanide m chlorophenil hydrazone (CCCP) causes decreased growth in yeast lacking SDO1. In fact, in normal cells, overexpression of BTN1 mirrors the effect of CCCP, with both resulting in increased vacuolar pH due to alterations in the coupling of V ATPase-dependent H(+) transport and ATP hydrolysis. Thus, we propose that Sdo1p and SBDS work to regulate Btn1p and CLN3, respectively. This report highlights a novel mechanism for controlling vacuole/lysosome homeostasis by the ribosome maturation pathway that may contribute to the cellular abnormalities associated with juvenile Batten disease and Shwachman-Bodian-Diamond syndrome. PMID- 20015956 TI - Relative tissue expression of homologous torsinB correlates with the neuronal specific importance of DYT1 dystonia-associated torsinA. AB - A three base-pair deletion in the widely expressed TOR1A gene causes the childhood onset, neurological disease of DYT1 dystonia. Mouse Tor1a gene knockout also specifically affects the developing nervous system. However, in both cases, the basis of neuronal tissue specificity is unknown. TorsinA is one of four predicted mammalian torsin ATPases associated with assorted cellular activities (AAA+) proteins, raising the possibility that expression of a functionally homologous torsin compensates for torsinA loss in non-neuronal tissues. We find that all four mammalian torsins are endoplasmic reticulum resident glycoproteins. TorsinA, torsinB and torsin2 are all present in large M(r) complexes, which suggests that each assembles into an oligomeric AAA+ enzyme. Introducing a mutation (WB(EQ)) that typically stabilizes AAA+ proteins in a substrate-bound state causes torsinA and torsinB to associate with a shared nuclear envelope (NE) binding partner and this NE localization requires the torsinA interacting protein, lamina associated polypeptide 1. Although torsin proteins are widely expressed in the adult mouse, we identified that embryonic neuronal tissues contain relatively low torsinB levels. Therefore, our results reveal that torsinB expression inversely correlates with the cell and developmental requirement for torsinA. In conclusion, multiple cell types appear to utilize torsin AAA+ proteins and differential expression of torsinB may contribute to both the neuronal specific importance of torsinA and the symptom specificity of DYT1 dystonia. PMID- 20015957 TI - Specific saposin C deficiency: CNS impairment and acid beta-glucosidase effects in the mouse. AB - Saposins A, B, C and D are derived from a common precursor, prosaposin (psap). The few patients with saposin C deficiency develop a Gaucher disease-like central nervous system (CNS) phenotype attributed to diminished glucosylceramide (GC) cleavage activity by acid beta-glucosidase (GCase). The in vivo effects of saposin C were examined by creating mice with selective absence of saposin C (C-/ ) using a knock-in point mutation (cysteine-to-proline) in exon 11 of the psap gene. In C-/- mice, prosaposin and saposins A, B and D proteins were present at near wild-type levels, but the saposin C protein was absent. By 1 year, the C-/- mice exhibited weakness of the hind limbs and progressive ataxia. Decreased neuromotor activity and impaired hippocampal long-term potentiation were evident. Foamy storage cells were observed in dorsal root ganglion and there was progressive loss of cerebellar Purkinje cells and atrophy of cerebellar granule cells. Ultrastructural analyses revealed inclusions in axonal processes in the spinal cord, sciatic nerve and brain, but no excess of multivesicular bodies. Activated microglial cells and astrocytes were present in thalamus, brain stem, cerebellum and spinal cord, indicating regional pro-inflammatory responses. No storage cells were found in visceral organs of these mice. The absence of saposin C led to moderate increases in GC and lactosylceramide (LacCer) and their deacylated analogues. These results support the view that saposin C has multiple roles in glycosphingolipid (GSL) catabolism as well as a prominent function in CNS and axonal integrity independent of its role as an optimizer/stabilizer of GCase. PMID- 20015958 TI - Loss of IGF2 imprinting is associated with abrogation of long-range intrachromosomal interactions in human cancer cells. AB - Nuclear architecture and chromatin geography are important factors in the regulation of gene expression, as these components may play a vital epigenetic role both in normal physiology as well as in the initiation and progression of malignancies. Using a modification of the chromosome conformation capture (3C) technique, we examined long-range chromatin interactions of the imprinted human IGF2 gene. We demonstrate that numerous intrachromosomal interactions occur along both parental alleles in normal tissues, where the IGF2 is paternally expressed, as well as in normal liver where gene expression is biallelic. Long-range and allele-specific interactions occur between the IGF2/H19 imprinting control region 1 (ICR1) and ICR2, a region which regulates an imprinted gene cluster nearly a megabase distant from IGF2. Loss of genomic imprinting is a common epigenetic event in cancer, and long-range interactions have not been examined in malignant cells. In cancer cell lines in which IGF2 imprinting is maintained (MOI), essentially all of the 3C interactions seen in normal cells were preserved. However, in cells in which IGF2 imprinting was lost (LOI), nearly all of the long range chromatin interactions involving IGF2 were abrogated. A three-dimensional computer model depicts the physical interactions between the IGF2 promoter and ICR1 in MOI cells, while the model of LOI lung cancer cells is flattened with few long-range interactions. This dramatic change in the three-dimension configuration of the chromatin at the IGF2 locus in LOI cancer cells suggests that the loss of imprinting may lead to a variety of changes in gene expression in addition to changes in IGF2 transcription. PMID- 20015959 TI - A cis-regulatory site downregulates PTHLH in translocation t(8;12)(q13;p11.2) and leads to Brachydactyly Type E. AB - Parathyroid hormone-like hormone (PTHLH) is an important chondrogenic regulator; however, the gene has not been directly linked to human disease. We studied a family with autosomal-dominant Brachydactyly Type E (BDE) and identified a t(8;12)(q13;p11.2) translocation with breakpoints (BPs) upstream of PTHLH on chromosome 12p11.2 and a disrupted KCNB2 on 8q13. We sequenced the BPs and identified a highly conserved Activator protein 1 (AP-1) motif on 12p11.2, together with a C-ets-1 motif translocated from 8q13. AP-1 and C-ets-1 bound in vitro and in vivo at the derivative chromosome 8 breakpoint [der(8) BP], but were differently enriched between the wild-type and BP allele. We differentiated fibroblasts from BDE patients into chondrogenic cells and found that PTHLH and its targets, ADAMTS-7 and ADAMTS-12 were downregulated along with impaired chondrogenic differentiation. We next used human and murine chondrocytes and observed that the AP-1 motif stimulated, whereas der(8) BP or C-ets-1 decreased, PTHLH promoter activity. These results are the first to identify a cis-directed PTHLH downregulation as primary cause of human chondrodysplasia. PMID- 20015960 TI - Traditional Medicine to Modern Pharmacogenomics: Ayurveda Prakriti Type and CYP2C19 Gene Polymorphism Associated with the Metabolic Variability. AB - Traditional Indian medicine-Ayurveda-classifies the human population into three major constituents or Prakriti known as Vata, Pitta and Kapha types. Earlier, we have demonstrated a proof of concept to support genetic basis for Prakriti. The descriptions in Ayurveda indicate that individuals with Pitta Prakriti are fast metabolizers while those of Kapha Prakriti are slow metabolizers. We hypothesized that different Prakriti may have different drug metabolism rates associated with drug metabolizing enzyme (DME) polymorphism. We did CYP2C19 (Phase I DME) genotyping in 132 unrelated healthy subjects of either sex by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) technique. We observed significant association between CYP2C19 genotype and major classes of Prakriti types. The extensive metabolizer (EM) genotype (*1/*1, *1/*2, *1/*3) was found to be predominant in Pitta Prakriti (91%). Genotype (*1/*3) specific for EM group was present only in Pitta Prakriti. Poor metabolizer (PM) genotype (*2/*2, *2/*3, *3/*3) was highest (31%) in Kapha Prakriti when compared with Vata (12%) and Pitta Prakriti (9%). Genotype (*2/*3) which is typical for PM group was significant in Kapha Prakriti (odds ratio = 3.5, P = .008). We observed interesting correlations between CYP2C19 genotypes and Prakriti with fast and slow metabolism being one of the major distinguishing and differentiating characteristics. These observations are likely to have significant impact on phenotype-genotype correlation, drug discovery, pharmacogenomics and personalized medicine. PMID- 20015961 TI - Infections and autoimmunity: the multifaceted relationship. AB - Multiple factors are thought to contribute to the development of immune response to self, including differences in genotypes, hormonal milieu, and environmental factors. This review focuses on the pivotal role of infection in the induction of autoimmune disorders. Although the development of autoimmune phenomena linked to infections is a common finding, the onset of autoimmune diseases is a rare event, arising from a combination of genetic susceptibility and environmental factors. There are several mechanisms through which pathogens can initiate or perpetuate autoimmunity. Some of them are antigen-specific, including molecular mimicry, expression of modified, cryptic, or new antigenic determinants, and superantigens. Others are nonspecific and collectively known as "bystander activation." They include enhanced processing and presentation of self-antigens, immune cell activation, cytokine release, and cell apoptosis/necrosis. Infections may also trigger organ-specific autoimmune diseases, but studies carried out until now have provided conflicting and inconclusive results regarding the role of viral and bacterial agents. Infections and autoimmune diseases have multifaceted and multidirectional relationships. It has been suggested recently that infections cannot only induce or precipitate autoimmune diseases, but they may also protect from autoimmunity or even abrogate an ongoing autoimmune process depending on the interaction between microorganisms and host. Therefore, we should look at microorganisms, not only as causes of infections but also as potential agents able to modulate the immune system. On the other hand, numerous evidences have emerged regarding the higher susceptibility of autoimmune patients to infections, possibly as a result of immunosuppressive therapy and treatment with biologic agents. PMID- 20015962 TI - Public concerns and behaviours towards solid waste management in Italy. AB - A self-administered questionnaire investigated knowledge, perceptions of the risks to health associated with solid waste management, and practices about waste management in a random sample of 1181 adults in Italy. Perceived risk of developing cancer due to solid waste burning was significantly higher in females, younger, with an educational level lower than university and who believed that improper waste management is linked to cancer. Respondents who had visited a physician at least once in the last year for fear of contracting a disease due to the non-correct waste management had an educational level lower than university, have modified dietary habits for fear of contracting disease due to improper waste management, believe that improper waste management is linked to allergies, perceive a higher risk of contracting infectious disease due to improper waste management and have participated in education/information activities on waste management. Those who more frequently perform with regularity differentiate household waste collection had a university educational level, perceived a higher risk of developing cancer due to solid waste burning, had received information about waste collection and did not need information about waste management. Educational programmes are needed to modify public concern about adverse health effects of domestic waste. PMID- 20015964 TI - Prevalence and characteristics of diabetes among homeless people attending shelters in Paris, France, 2006. AB - Chronic diseases are frequent in homeless people. The aims of this study were to estimate the prevalence of diabetes in people living in shelters and to describe the characteristics of homeless people previously diagnosed with diabetes. Diabetes screening was systematically performed in nine shelters in Paris, from October to December 2006. We found a high prevalence of diabetes comparable with those of the general population, but a high frequency of major complications, in spite of a relatively young age, short duration since diabetes diagnosis and moderate glycaemic control. This study underlines the burden of diabetes among precarious people and supports the development of more effective strategies to improve diabetes management in this population, especially regarding podiatric care. PMID- 20015965 TI - Comparing actual to estimated base rates of "abnormal" scores on neuropsychological test batteries: implications for interpretation. AB - Clinicians can use the prevalence of low scores to help interpret test performance. However, this information is limited for most test batteries. In 2007, Crawford, Garthwaite, and Gault presented Monte Carlo simulation software for estimating the base rates of low scores for any battery of tests. The purpose of this study is to examine the accuracy of a Monte Carlo simulation program for estimating the base rates of low scores. Base rates of low scores were: (a) calculated from large normative samples (actual base rates) for the Neuropsychological Assessment Battery and the Wechsler Adult Intelligence Scale III/Wechsler Memory Scale-Third Edition and compared to (b) Monte Carlo estimations (estimated base rates). Monte Carlo estimations of the base rates of low scores had good accuracy when compared with the actual base rates of low scores for the two batteries. However, estimated base rates lose considerable accuracy in those with low or high intelligence. Monte Carlo simulation software is a potential option for clinicians to compute the base rates of low scores for any battery with published intercorrelations. However, the Monte Carlo program underestimates the base rates for those with low intelligence and overestimates the base rates for those with high intelligence. PMID- 20015966 TI - Neuropsychological features of indigent murder defendants and death row inmates in relation to homicidal aspects of their crimes. AB - Neuropsychological features of 77 indigent murder defendants and death row inmates were examined in relation to criminal variables underlying their homicidal acts. Clinically, the sample was characterized by elevated rates of developmental disorders (49%), personality disorders (54%), Axis I psychiatric disorders (45%), substance abuse (86%), and history of violence (43%). By statute, killing more than one person is an aggravating factor in many jurisdictions that renders a murder defendant eligible for the death penalty. Individuals who committed a single murder were characterized by executive dysfunction, lower intelligence, slower speed of information processing, and a higher frequency of developmental disorders (58%), relative to those charged and/or convicted of killing two or more people, who were characterized by a significantly higher rate of personality disorders (79%) and a lower rate of developmental disorders (34%). Additionally, using the FBI criminal classification system for categorizing homicide by motive, a significant difference in the frequency of psychosis was found among subgroups associated with the following motives: Criminal enterprise; personal cause; and sex. The collective neuropsychological profile of the sample revealed that executive functions were significantly decreased, relative to memory functions, with over half of the sample (55%) demonstrating executive dysfunction. PMID- 20015967 TI - Reproductive isolation between 2 karyotypes in natural populations of the leaf beetle Chrysolina aurichalcea. AB - Japanese populations of the leaf beetle, Chrysolina aurichalcea, have 2 main karyotypes, Type 31 and Type 41, which greatly differ in chromosome number and form. We examined their geographical distribution and reproductive isolation in southern Hokkaido, northern Japan. Type 41 was widely distributed in the survey area, whereas Type 31 was restricted to a narrow portion. A transect across each karyotypic habitat revealed that both karyotypes co-occurred in a approximately 5 km wide zone, with few hybrids or hybrid derivatives. Assessment of the fine scale distribution showed that the 2 karyotypes do not occupy separate microhabitats within the sympatric zone. Karyotype combinations of copulating pairs in the sympatric zone showed no evidence of assortative mating according to karyotype. These findings suggest that postmating isolation between the 2 karyotypes is strong, whereas premating isolation is weak. It does not agree with results from previous studies, which reported a substantial degree of both post- and premating isolation between the karyotypes. Possible causes of the discrepancy in premating isolation are discussed. PMID- 20015968 TI - Differential binding of Escherichia coli McrA protein to DNA sequences that contain the dinucleotide m5CpG. AB - The Escherichia coli McrA protein, a putative C(5)-methylcytosine/C(5)-hydroxyl methylcytosine-specific nuclease, binds DNA with symmetrically methylated HpaII sequences (Cm5CGG), but its precise recognition sequence remains undefined. To determine McrA's binding specificity, we cloned and expressed recombinant McrA with a C-terminal StrepII tag (rMcrA-S) to facilitate protein purification and affinity capture of human DNA fragments with m5C residues. Sequence analysis of a subset of these fragments and electrophoretic mobility shift assays with model methylated and unmethylated oligonucleotides suggest that N(Y > R) m5CGR is the canonical binding site for rMcrA-S. In addition to binding HpaII-methylated double-stranded DNA, rMcrA-S binds DNA containing a single, hemimethylated HpaII site; however, it does not bind if A, C, T or U is placed across from the m5C residue, but does if I is opposite the m5C. These results provide the first systematic analysis of McrA's in vitro binding specificity. PMID- 20015969 TI - Functional interplay between chromatin remodeling complexes RSC, SWI/SNF and ISWI in regulation of yeast heat shock genes. AB - Chromatin remodeling is an essential part of transcription initiation. We show that at heat shock gene promoters functional interactions between individual ATP dependent chromatin remodeling complexes play critical role in both nucleosome displacement and Pol II recruitment. Using HSP12, HSP82 and SSA4 gene promoters as reporters, we demonstrated that while inactivation of SNF2, a critical ATPase of the SWI/SNF complex, primarily affects the HSP12 promoter, depletion of STH1- a SNF2 homolog from the RSC complex reduces histone displacement and abolishes the Pol II recruitment at all three promoters. From these results, we conclude that redundancy between SWI/SNF and RSC complexes is only partial and likely is affecting different chromatin remodeling steps. While inactivation of other individual ATP-dependent chromatin remodeling complexes negligibly affects reporter promoters, combinatorial inactivation of SNF2 and ISW1 has a synergistic effect by diminishing histone loss during heat induction and eliminating Pol II recruitment. Importantly, it also eliminates preloading of HSF on HSP82 and SSA4 promoters before heat shock and diminishes HSF binding during heat shock. These observations suggest that prior action of chromatin remodeling complexes is necessary for the activator binding. PMID- 20015970 TI - The Sanger FASTQ file format for sequences with quality scores, and the Solexa/Illumina FASTQ variants. AB - FASTQ has emerged as a common file format for sharing sequencing read data combining both the sequence and an associated per base quality score, despite lacking any formal definition to date, and existing in at least three incompatible variants. This article defines the FASTQ format, covering the original Sanger standard, the Solexa/Illumina variants and conversion between them, based on publicly available information such as the MAQ documentation and conventions recently agreed by the Open Bioinformatics Foundation projects Biopython, BioPerl, BioRuby, BioJava and EMBOSS. Being an open access publication, it is hoped that this description, with the example files provided as Supplementary Data, will serve in future as a reference for this important file format. PMID- 20015971 TI - Definition of the interacting interfaces of Apobec3G and HIV-1 Vif using MAPPIT mutagenesis analysis. AB - The host restriction factor Apobec3G is a cytidine deaminase that incorporates into HIV-1 virions and interferes with viral replication. The HIV-1 accessory protein Vif subverts Apobec3G by targeting it for proteasomal degradation. We propose a model in which Apobec3G N-terminal domains symmetrically interact via a head-to-head interface containing residues 122 RLYYFW 127. To validate this model and to characterize the Apobec3G-Apobec3G and the Apobec3G-Vif interactions, the mammalian protein-protein interaction trap two-hybrid technique was used. Mutations in the head-to-head interface abrogate the Apobec3G-Apobec3G interaction. All mutations that inhibit Apobec3G-Apobec3G binding also inhibit the Apobec3G-Vif interaction, indicating that the head-to head interface plays an important role in the interaction with Vif. Only the D128K, P129A and T32Q mutations specifically affect the Apobec3G-Vif association. In our model, D128, P129 and T32 cluster at the edge of the head-to-head interface, possibly forming a Vif binding site composed of two Apobec3G molecules. We propose that Vif either binds at the Apobec3G head-to-head interface or associates with an RNA-stabilized Apobec3G oligomer. PMID- 20015972 TI - PANTHER version 7: improved phylogenetic trees, orthologs and collaboration with the Gene Ontology Consortium. AB - Protein Analysis THrough Evolutionary Relationships (PANTHER) is a comprehensive software system for inferring the functions of genes based on their evolutionary relationships. Phylogenetic trees of gene families form the basis for PANTHER and these trees are annotated with ontology terms describing the evolution of gene function from ancestral to modern day genes. One of the main applications of PANTHER is in accurate prediction of the functions of uncharacterized genes, based on their evolutionary relationships to genes with functions known from experiment. The PANTHER website, freely available at http://www.pantherdb.org, also includes software tools for analyzing genomic data relative to known and inferred gene functions. Since 2007, there have been several new developments to PANTHER: (i) improved phylogenetic trees, explicitly representing speciation and gene duplication events, (ii) identification of gene orthologs, including least diverged orthologs (best one-to-one pairs), (iii) coverage of more genomes (48 genomes, up to 87% of genes in each genome; see http://www.pantherdb.org/panther/summaryStats.jsp), (iv) improved support for alternative database identifiers for genes, proteins and microarray probes and (v) adoption of the SBGN standard for display of biological pathways. In addition, PANTHER trees are being annotated with gene function as part of the Gene Ontology Reference Genome project, resulting in an increasing number of curated functional annotations. PMID- 20015973 TI - Nicotine-patch therapy on mucocutaneous lesions of Behcet's disease: a case series. AB - OBJECTIVE: We report the use of nicotine-patch therapy on active mucocutaneous lesions of Behcet's disease (BD). METHODS: Five BD ex-smoker patients with refractory active mucocutaneous manifestations were treated with nicotine patches for 6 months. RESULTS: Four out of five patients quickly responded to nicotine patch therapy and experienced a complete regression of mucocutaneous lesions. Other manifestations of BD did not respond and new manifestations appeared during this treatment. One patient had no benefit from therapy but on restarting smoking it was promptly effective. CONCLUSIONS: Mucocutaneous lesions associated with BD may be modulated by smoking. Both smoking and nicotine-replacement therapy may be efficacious not only on oral aphthae, but also on other mucocutaneous manifestations, whereas the efficacy in the treatment and prevention of other systemic manifestations of BD is not proven. At least in ex-smokers, nicotine in its pure form is well tolerated and its use could be justified in selected cases of BD with predominant and recurrent refractory mucocutaneous manifestations. PMID- 20015974 TI - Long-term outcome of systemic sclerosis-associated pulmonary arterial hypertension treated with bosentan as first-line monotherapy followed or not by the addition of prostanoids or sildenafil. AB - OBJECTIVE: Data on long-term efficacy of bosentan, an oral dual ET receptor antagonist, in SSc-associated pulmonary arterial hypertension (SSc-PAH) are lacking. We aimed to describe the long-term outcome of SSc-PAH treated with first line monotherapy bosentan followed or not by the addition of prostanoids or sildenafil. METHODS: A prospective analysis of 49 consecutive SSc-PAH patients treated with first-line bosentan was performed. New York Heart Association (NYHA) functional class, 6-min walk distance (6MWD) and haemodynamics were assessed at baseline and after 4 and 12 months. RESULTS: At 4 months, significant improvements in NYHA functional class and haemodynamics were observed with stabilization at 1 year. There was no significant improvement in 6MWD. Overall survival estimates were 80, 56 and 51% at 1, 2 and 3 years, respectively, and were significantly worse than those in a cohort of patients with idiopathic PAH (92, 89 and 79% at 1, 2 and 3 years, respectively; P < 0.0001). Twenty-three patients (47%) died after a mean follow-up of 23 (18) months. In multivariate analysis, baseline and 4-month NYHA functional class and 4-month cardiac index were independent factors associated with overall survival. CONCLUSIONS: In our cohort of consecutive SSc-PAH patients treated with first-line bosentan, improvement in NYHA functional class and haemodynamics was significant after 4 months of treatment and stabilized afterwards. One-year overall survival rate was higher than previously reported in historical series. However, long-term prognosis remains poor. Our study underlines the importance of haemodynamic evaluation 4 months after the start of treatment to provide strong parameters associated with survival-like cardiac index. PMID- 20015975 TI - Lateral back pain identifies prevalent vertebral fractures in post-menopausal women: cross-sectional analysis of a primary care-based cohort. AB - OBJECTIVE: Vertebral fractures (VFs) are frequently under-recognized, reflecting their lack of diagnostic clinical features. For example, although VFs are associated with back pain, this is also common in the general population. To establish whether back pain can be used to recognize patients with VF, we investigated the site of pain in people with and without VFs using a simple tool. METHODS: A cohort of 504 post-menopausal women was recruited from primary care in South West UK. Back pain was assessed by self-completion of the Margolis pain diagram, and analysis was modified to assess whether pain was mid-line or lateral. VFs were diagnosed by the algorithm-based qualitative method on radiographs. A cross-sectional analysis was carried out to assess the association between back pain and VFs. RESULTS: Three hundred and twenty-two women (64.1%) reported back pain over the last 12 months. Thirty seven (7.3%) had one or more VFs. In women with back pain, the presence of lateral waist area pain was associated with a 4.5-fold increased risk of VFs [odds ratio (OR) 4.48; 95% CI 2.02, 9.94; P < 0.001]. CONCLUSIONS: In post-menopausal women with back pain, the presence of lateral waist pain, as shown on the Margolis pain diagram, may identify women at higher risk of prevalent VF. PMID- 20015976 TI - The RIG-I-like receptor IFIH1/MDA5 is a dermatomyositis-specific autoantigen identified by the anti-CADM-140 antibody. AB - OBJECTIVES: Various autoantibodies are detected in the sera of PM/DM patients. Some of them are specific to PM/DM patients and closely associated with clinical manifestations of the diseases. Recently, the anti-CADM-140 antibody was reported to be found specifically in clinically amyopathic DM (C-ADM) patients and to be associated with acute interstitial lung disease (ILD). We assessed the clinical significance of the anti-CADM-140 antibody and then investigated the autoantigen recognized by the anti-CADM-140 antibody. METHODS: Autoantibodies were screened in 192 patients with various CTDs and 21 healthy controls using immunoprecipitation with [(35)S]methionine-labelled HeLa cells. Immunoabsorbent column chromatography was used to purify an autoantigen that was subsequently subjected to peptide mass fingerprinting. RESULTS: The anti-CADM-140 antibody was revealed to be specific to DM. Most of the anti-CADM-140-positive patients were C ADM although some of them showed apparent myositis. The anti-CADM-140-positive patients frequently showed hyperferritinaemia and acute progressive ILD with poor prognosis. The anti-CADM-140 antibody was shown to recognize IFN induced with helicase C domain protein 1 (IFIH1), also known as the melanoma differentiation associated gene 5 (MDA5), which is one of the RIG-I-like receptors and plays a role in innate immune responses. CONCLUSION: The anti-CADM-140 antibody was a marker of DM and intractable ILD and recognized IFIH1/MDA5, which is involved in innate immunity. These findings may give a new insight into the pathogenesis of DM. PMID- 20015977 TI - Extracellular ubiquitin inhibits beta-AR-stimulated apoptosis in cardiac myocytes: role of GSK-3beta and mitochondrial pathways. AB - AIMS: Beta-adrenergic receptor (beta-AR) stimulation induces apoptosis in adult rat ventricular myocytes (ARVMs) via the activation of glycogen synthase kinase 3beta (GSK-3beta) and mitochondrial pathways. However, beta-AR stimulation induces apoptosis only in a fraction ( approximately 15-20%) of ARVMs. We hypothesized that ARVMs may secrete/release a survival factor(s) which protects 80-85% of cells from apoptosis. METHODS AND RESULTS: Using two-dimensional gel electrophoresis followed by MALDI TOF and MS/MS, we identified ubiquitin (Ub) in the conditioned media of ARVMs treated with beta-AR agonist (isoproterenol). Western blot analysis confirmed increased Ub levels in the conditioned media 3 and 6 h after beta-AR stimulation. Inhibition of beta1-AR and beta2-AR subtypes inhibited beta-AR-stimulated increases in extracellular levels of Ub, whereas activation of adenylyl cyclase using forskolin mimicked the effects of beta-AR stimulation. Incubation of cells with exogenous biotinylated Ub followed by western blot analysis of the cell lysates showed uptake of extracellular Ub into cells, which was found to be higher after beta-AR stimulation (1.9 +/- 0.4-fold; P < 0.05 vs. control, n = 6). Pre-treatment with Ub inhibited beta-AR-stimulated increases in apoptosis. Inhibition of phosphoinositide 3-kinase using wortmannin and LY-294002 prevented anti-apoptotic effects of extracellular Ub. Ub pre treatment inhibited beta-AR-stimulated activation of GSK-3beta and c-Jun N terminal kinase (JNK) and increases in the levels of cytosolic cytochrome c. The use of methylated Ub suggested that the anti-apoptotic effects of extracellular Ub are mediated via monoubiquitination. CONCLUSION: beta-AR stimulation increases levels of Ub in the conditioned media. Extracellular Ub plays a protective role in beta-AR-stimulated apoptosis, possibly via the inactivation of GSK-3beta/JNK and mitochondrial pathways. PMID- 20015978 TI - Novel functions of the alphavirus nonstructural protein nsP3 C-terminal region. AB - The functions of the alphavirus-encoded nonstructural protein nsP3 during infection are poorly understood. In contrast, nsP1, nsP2, and nsP4 have known enzymatic activities and functions. A functional analysis of the C-terminal region of nsP3 of Semliki Forest virus revealed the presence of a degradation signal that overlaps with a sequence element located between nsP3 and nsP4 that is required for proteolytic processing. This element was responsible for the short half-life (1 h) of individually expressed nsP3, and it also was functionally transferable to other proteins. Inducible cell lines were used to express native nsP3 or truncated mutants. The removal of 10 C-terminal amino acid (aa) residues from nsP3 increased the half-life of the protein approximately 8 fold. While the deletion of 30 C-terminal aa residues resulted in a similar stabilization, this deletion also changed the cellular localization of nsP3. This truncated mutant no longer exhibited a punctate localization in the cytoplasm, but instead filamentous stretches could be formed around the nuclei of induced cells, suggesting the existence of an additional functional element upstream of the degradation signal. C-terminally truncated uncleavable polyprotein P12(CA)3del30 was localized diffusely, which is in contrast to P12(CA)3, which is known to be associated with vesicle membranes. The induction of nsP3 or its truncated forms reduced the efficiency of virus multiplication in corresponding cells by affecting different steps of the infection cycle. The expression of nsP3 or a mutant lacking the 10 C-terminal aa residues repressed the establishment of infection, while the expression of nsP3 lacking 30 C-terminal aa residues led to the reduced synthesis of subgenomic RNA. PMID- 20015979 TI - RNA-dependent RNA polymerase 6 delays accumulation and precludes meristem invasion of a viroid that replicates in the nucleus. AB - The detection of viroid-derived small RNAs (vd-sRNAs) similar to the small interfering RNAs (siRNAs, 21 to 24 nucleotides [nt]) in plants infected by nuclear-replicating members of the family Pospiviroidae (type species, Potato spindle tuber viroid [PSTVd]) indicates that they are inducers and targets of the RNA-silencing machinery of their hosts. RNA-dependent RNA polymerase 6 (RDR6) catalyzes an amplification circuit producing the double-stranded precursors of secondary siRNAs. Recently, the role of RDR6 in restricting systemic spread of certain RNA viruses and precluding their invasion of the apical growing tip has been documented using RDR6-silenced Nicotiana benthamiana (NbRDR6i) plants. Here we show that RDR6 is also engaged in regulating PSTVd levels: accumulation of PSTVd genomic RNA was increased in NbRDR6i plants with respect to the wild-type controls (Nbwt) early in infection, whereas this difference decreased or disappeared in later infection stages. Moreover, in situ hybridization revealed that RDR6 is involved in restricting PSTVd access in floral and vegetative meristems, thus providing firm genetic evidence for an antiviroid RNA silencing mechanism. RNA gel blot hybridization and deep sequencing showed in wt and RDR6i backgrounds that PSTVd sRNAs (i) accumulate to levels paralleling their genomic RNA, (ii) display similar patterns with prevailing 22- or 21-nt plus-strand species, and (iii) adopt strand-specific hot spot profiles along the genomic RNA. Therefore, the surveillance mechanism restraining entry of some RNA viruses into meristems likely also controls PSTVd access in N. benthamiana. Unexpectedly, deep sequencing also disclosed in NbRDR6i plants a profile of RDR6-derived siRNA dominated by 21-nt plus-strand species mapping within a narrow window of the hairpin RNA stem expressed transgenically for silencing RDR6, indicating that minus-strand siRNAs silencing the NbRDR6 mRNA represent a minor fraction of the total siRNA population. PMID- 20015980 TI - Combined administration with DNA encoding vesicular stomatitis virus G protein enhances DNA vaccine potency. AB - DNA vaccines have recently emerged at the forefront of approaches to harness the immune system in the prevention and treatment of viral infections, as well as the prevention and treatment of cancers. However, these vaccines suffer from limited efficacy since they often fail to produce significant antigen-specific CD8(+) T cell responses. We report here a novel concept for DNA vaccine design that exploits the unique and powerful ability of viral fusogenic membrane glycoproteins (FMGs) to couple concentrated antigen transfer to dendritic cells (DCs) with local induction of the acute inflammatory response. Intramuscular administration into mice by electroporation technology of a plasmid containing the FMG gene from vesicular stomatitis virus (VSV-G)-together with DNA encoding the E7 protein of human papillomavirus type 16, a model cervical cancer antigen elicited robust E7-specific CD8(+) T-cell responses, as well as therapeutic control of E7-expressing tumors. This effect could potentially be mediated through the immunogenic form of cellular fusion and necrosis induced by VSV-G, which in a concerted fashion provokes leukocyte infiltration into the inoculation site, enhances cross-presentation of antigen to DCs, and stimulates them to mature efficiently. Thus, the incorporation of FMGs into DNA vaccines holds promise for the successful control of viral infections and cancers in the clinic. PMID- 20015981 TI - Inhibition of sterol biosynthesis reduces tombusvirus replication in yeast and plants. AB - The replication of plus-strand RNA viruses depends on subcellular membranes. Recent genome-wide screens have revealed that the sterol biosynthesis genes ERG25 and ERG4 affected the replication of Tomato bushy stunt virus (TBSV) in a yeast model host. To further our understanding of the role of sterols in TBSV replication, we demonstrate that the downregulation of ERG25 or the inhibition of the activity of Erg25p with an inhibitor (6-amino-2-n-pentylthiobenzothiazole; APB) leads to a 3- to 5-fold reduction in TBSV replication in yeast. In addition, the sterol biosynthesis inhibitor lovastatin reduced TBSV replication by 4-fold, confirming the importance of sterols in viral replication. We also show reduced stability for the p92(pol) viral replication protein as well as a decrease in the in vitro activity of the tombusvirus replicase when isolated from APB-treated yeast. Moreover, APB treatment inhibits TBSV RNA accumulation in plant protoplasts and in Nicotiana benthamiana leaves. The inhibitory effect of APB on TBSV replication can be complemented by exogenous stigmasterol, the main plant sterol, suggesting that sterols are required for TBSV replication. The silencing of SMO1 and SMO2 genes, which are orthologs of ERG25, in N. benthamiana reduced TBSV RNA accumulation but had a lesser inhibitory effect on the unrelated Tobacco mosaic virus, suggesting that various viruses show different levels of dependence on sterol biosynthesis for their replication. PMID- 20015982 TI - A systemic neutrophil response precedes robust CD8(+) T-cell activation during natural respiratory syncytial virus infection in infants. AB - Severe primary respiratory syncytial virus (RSV) infections are characterized by bronchiolitis accompanied by wheezing. Controversy exists as to whether infants suffer from virus-induced lung pathology or from excessive immune responses. Furthermore, detailed knowledge about the development of primary T-cell responses to viral infections in infants is lacking. We studied the dynamics of innate neutrophil and adaptive T-cell responses in peripheral blood in relation to the viral load and parameters of disease in infants admitted to the intensive care unit with severe RSV infection. Analysis of primary T-cell responses showed substantial CD8(+) T-cell activation, which peaked during convalescence. A strong neutrophil response, characterized by mobilization of bone marrow-derived neutrophil precursors, preceded the peak in T-cell activation. The kinetics of this neutrophil response followed the peak of clinical symptoms and the viral load with a 2- to 3-day delay. From the sequence of events, we conclude that CD8(+) T-cell responses, initiated during primary RSV infections, are unlikely to contribute to disease when it is most severe. The mobilization of precursor neutrophils might reflect the strong neutrophil influx into the airways, which is a characteristic feature during RSV infections and might be an integral pathogenic process in the disease. PMID- 20015983 TI - Induction of protective immunity against murine gammaherpesvirus 68 infection in the absence of viral latency. AB - Human gammaherpesviruses, Epstein-Barr virus, and human herpesvirus 8/Kaposi's sarcoma-associated herpesvirus are important pathogens associated with diseases, including lymphomas and other malignancies. Murine gammaherpesvirus 68 (MHV-68) is used as an experimental model system to study the host immune control of infection and explore novel vaccine strategies based on latency-deficient live viruses. We studied the properties and the potential of a recombinant MHV-68 (AC RTA) in which the genes required for persistent infection were replaced by a constitutively expressed viral transcription activator, RTA, which dictates the virus to lytic replication. After intranasal infection of mice, replication of AC RTA in the lung was attenuated, and no AC-RTA virus or viral DNA was detected in the isolated splenocytes, indicating a lack of latency in the spleen. Infection of the AC-RTA virus elicited both cellular immune responses and virus-specific IgG at a level comparable to that elicited by infection of the wild-type virus. Importantly, vaccination of AC-RTA was able to protect mice against subsequent challenge by the wild-type MHV-68. AC-RTA provides a vaccine strategy for preventing infection of human gammaherpesviruses. Furthermore, our results suggest that immunity to the major latent antigens is not required for protection. PMID- 20015984 TI - Compartmentalization and clonal amplification of HIV-1 variants in the cerebrospinal fluid during primary infection. AB - Human immunodeficiency virus type 1 (HIV-1)-associated dementia (HAD) is a severe neurological disease that affects a subset of HIV-1-infected individuals. Increased compartmentalization has been reported between blood and cerebrospinal fluid (CSF) HIV-1 populations in subjects with HAD, but it is still not known when compartmentalization arises during the course of infection. To assess HIV-1 genetic compartmentalization early during infection, we compared HIV-1 populations in the peripheral blood and CSF in 11 primary infection subjects, with analysis of longitudinal samples over the first 18 months for a subset of subjects. We used heteroduplex tracking assays targeting the variable regions of env and single-genome amplification and sequence analysis of the full-length env gene to identify CSF-compartmentalized variants and to examine viral genotypes within the compartmentalized populations. For most subjects, HIV-1 populations were equilibrated between the blood and CSF compartments. However, compartmentalized HIV-1 populations were detected in the CSF of three primary infection subjects, and longitudinal analysis of one subject revealed that compartmentalization during primary HIV-1 infection was resolved. Clonal amplification of specific HIV-1 variants was identified in the CSF population of one primary infection subject. Our data show that compartmentalization can occur in the central nervous system (CNS) of subjects in primary HIV-1 infection in part through persistence of the putative transmitted parental variant or via viral genetic adaptation to the CNS environment. The presence of distinct HIV-1 populations in the CSF indicates that independent HIV-1 replication can occur in the CNS, even early after HIV-1 transmission. PMID- 20015985 TI - Regulation of porcine endogenous retrovirus release by porcine and human tetherins. AB - The risk of transmission of porcine endogenous retrovirus (PERV) is one of the major safety issues in xenotransplantation. Human tetherin, recently described as an antiviral protein able to inhibit the release of enveloped viruses, and its porcine homologue were shown to inhibit PERV release from producer cells, establishing themselves as candidate molecules to suppress PERV production in porcine xenografts by animal engineering. PMID- 20015986 TI - ICP27 phosphorylation site mutants display altered functional interactions with cellular export factors Aly/REF and TAP/NXF1 but are able to bind herpes simplex virus 1 RNA. AB - Herpes simplex virus 1 (HSV-1) protein ICP27 is a multifunctional regulatory protein that is phosphorylated. Phosphorylation can affect protein localization, protein interactions, and protein function. The major sites of ICP27 that are phosphorylated are serine residues 16 and 18, within a CK2 site adjacent to a leucine-rich region required for ICP27 export, and serine 114, within a PKA site in the nuclear localization signal. Viral mutants bearing serine-to-alanine or glutamic acid substitutions at these sites are defective in viral replication and gene expression. To determine which interactions of ICP27 are impaired, we analyzed the subcellular localization of ICP27 and its colocalization with cellular RNA export factors Aly/REF and TAP/NXF1. In cells infected with phosphorylation site mutants, ICP27 was confined to the nucleus even at very late times after infection. ICP27 did not colocalize with Aly/REF or TAP/NXF1, and overexpression of TAP/NXF1 did not promote the export of ICP27 to the cytoplasm. However, in vitro binding experiments showed that mutant ICP27 was able to bind to the same RNA substrates as the wild type. Nuclear magnetic resonance (NMR) analysis of the N terminus of ICP27 from amino acids 1 to 160, compared to mutants with triple substitutions to alanine or glutamic acid, showed that the mutations affected the overall conformation of the N terminus, such that mutant ICP27 was more flexible and unfolded. These results indicate that these changes in the structure of ICP27 altered in vivo protein interactions that occur in the N terminus but did not prevent RNA binding. PMID- 20015987 TI - Comparative immunogenicity of subtype a Human Immunodeficiency Virus type 1 envelope exhibiting differential exposure of conserved neutralization epitopes. AB - Development of broadly cross-reactive neutralizing antibodies (NAbs) remains a major goal of HIV-1 vaccine development, but most candidate envelope immunogens have had limited ability to cross-neutralize heterologous strains. To evaluate the immunogenicity of subtype A variants of HIV-1, rabbits were immunized with pairs of closely related subtype A envelopes from the same individual. In each immunogen pair, one variant was readily neutralized by a variety of monoclonal antibodies and plasma antibodies, while the other was neutralization resistant, suggesting differences in the exposures of key epitopes. The breadth of the antibody response was evaluated against subtype A, B, C, and D variants of HIV-1. The specificity of the immunogen-derived neutralizing antibody response was also compared to that of the infected individuals from whom these variants were cloned. None of the immunogens produced broad neutralizing antibodies in immunized animals, and most of the neutralizing antibodies were directed to the variable loops, particularly the V3 loop. No detectable antibodies to either of the potentially exposed conserved epitopes, the membrane proximal external region, or the CD4 binding site were found with immunized rabbits. In contrast, relatively little of the neutralizing activity within the plasma samples of the infected individuals was directed to linear epitopes within the variable loops. These data indicate that immunogens designed to expose conserved regions did not enhance generation of broadly neutralizing antibodies in comparison with the immunogens that failed to expose those regions using this immunization approach. PMID- 20015988 TI - Augmented latent membrane protein 1 expression from Epstein-Barr virus episomes with minimal terminal repeats. AB - The major oncogene of the Epstein-Barr virus (EBV), latent membrane protein 1 (LMP1), can be expressed from either of two promoters, ED-L1 or L1-TR, producing mRNAs of 2.8 kb or 3.5 kb, respectively. L1-TR, active in nasopharyngeal carcinoma and Hodgkin's lymphoma, is located within the first of a highly variable reiteration of terminal repeat (TR) sequences that are joined by random recombination upon circularization of the linear genome at entry into cells. To determine whether the resultant TR number affects LMP1 promoter activity, we isolated single-cell clones bearing episomes of distinct TR numbers (6TR to 12TR) from epithelial cells newly infected with EBV. LMP1 mRNA levels correlated directly with the quantity of LMP1 protein expressed but varied inversely to TR number. Unexpectedly, the 3.5-kb transcript predominated only at lower TR reiterations. Diminished L1-TR activity in the context of a higher TR count was confirmed with a green fluorescent protein (GFP) reporter construct driven by L1 TR. Various levels of LMP1, expressed from virus isogenic in all but TR number, produced divergent morphological and growth phenotypes in each cell clone. Abundant LMP1 in 6TR cells yielded a relatively cytostatic state compared to the proliferative one produced by intermediate and smaller amounts in 8TR and 12TR clones. These findings suggest that the diversification of TR number, inherent in a round of EBV reactivation and reinfection, may itself be a component of the oncogenic process. The replicative burst preceding onset of many EBV-linked cancers may increase the likelihood that LMP1 levels compatible with clonal outgrowth are achieved in a subset of infected cells. PMID- 20015989 TI - Testing the balanced electrostatic interaction hypothesis of hepatitis B virus DNA synthesis by using an in vivo charge rebalance approach. AB - Previously, a charge balance hypothesis was proposed to explain hepatitis B virus (HBV) capsid stability, assembly, RNA encapsidation, and DNA replication. This hypothesis emphasized the importance of a balanced electrostatic interaction between the positive charge from the arginine-rich domain (ARD) of the core protein (HBc) and the negative charge from the encapsidated nucleic acid. It remains unclear if any of the negative charge involved in this electrostatic interaction could come from the HBc protein per se, in addition to the encapsidated nucleic acid. HBc ARD IV mutant 173GG and ARD II mutant 173RR/R157A/R158A are arginine deficient and replication defective. Not surprisingly, the replication defect of ARD IV mutant 173GG can be rescued by restoring positively charged amino acids at the adjacent positions 174 and 175. However, most interestingly, it can be at least partially rescued by reducing negatively charged residues in the assembly domain, such as by glutamic acid-to alanine (E-to-A) substitutions at position 46 or 117 and to a much lesser extent at position 113. Similar results were obtained for ARD II mutant 173RR/R157A/R158A. These amino acids are located on the inner surfaces of HBc icosahedral particles, and their acidic side chains point toward the capsid interior. For HBV DNA synthesis, the relative amount of positive versus negative charge in the electrostatic interactions is more important than the absolute amount of positive or negative charge. These results support the concept that balanced electrostatic interaction is important during the viral life cycle. PMID- 20015990 TI - Xenotropic murine leukemia virus-related virus establishes an efficient spreading infection and exhibits enhanced transcriptional activity in prostate carcinoma cells. AB - Xenotropic murine leukemia virus-related virus (XMRV) is a novel human gammaretrovirus discovered in association with human prostate tumors. XMRV was first identified in prostate stromal cells surrounding the tumors of patients carrying a mutation in the HPC1 gene locus. To determine the tropism of XMRV in cell culture, we tested the ability of XMRV to spread and replicate in various prostate and nonprostate cell lines. We found that although the expression of XMRV viral proteins and the spread of infectious virus were minimal in a variety of cell lines, XMRV displayed robust expression and infection in LNCaP prostate tumor cells. The transcriptional activity of the XMRV long terminal repeat (LTR) was found to be higher than the Moloney murine leukemia virus LTRs in both LNCaP and WPMY-1 (simian virus 40-transformed prostate stromal cells). The U3 promoter of XMRV and a glucocorticoid response element (GRE) within the U3 were required for the transcriptional activity in LNCaP cells. Coexpression of the androgen receptor and stimulation with dihydrotestosterone stimulated XMRV-LTR-dependent transcription in 293T cells, and the GRE was required for this activity. These data suggest that XMRV may replicate more efficiently in LNCaP cells in part due to the transcriptional environment in LNCaP cells. PMID- 20015991 TI - ICP27 phosphorylation site mutants are defective in herpes simplex virus 1 replication and gene expression. AB - Herpes simplex virus 1 (HSV-1) protein ICP27 is a multifunctional regulatory protein that is posttranslationally modified by phosphorylation during viral infection. ICP27 has been shown to be phosphorylated on three serine residues, specifically serine residues 16 and 18, which are within casein kinase 2 (CK2) sites, and serine residue 114, which is within a protein kinase A (PKA) site. Phosphorylation is an important regulatory mechanism that is reversible and controls many signaling pathways, protein-protein interactions, and protein subcellular localization. To determine the role of phosphorylation in modulating the activities of ICP27, we constructed phosphorylation site mutations at each of the three serine residues. Single, double, and triple viral mutants were created in which alanine or glutamic acid was substituted for serines 16, 18, and 114. ICP27 phosphorylation site mutants were defective in viral replication and viral gene expression. Notably, ICP4-containing replication compartment formation was severely compromised, with the appearance of small ring-like structures that persisted even at late times after infection. Neither the colocalization of ICP27 with RNA polymerase II nor the formation of Hsc70 nuclear foci was observed during infection with the phosphorylation site mutants, both of which occur during wild-type HSV-1 infection. These data indicate that several key events in which ICP27 plays a role are curtailed during infection with ICP27 phosphorylation site mutants. PMID- 20015993 TI - Stability of murine cytomegalovirus genome after in vitro and in vivo passage. AB - While large DNA viruses are thought to have low mutation rates, only a small fraction of their genomes have been analyzed at the single-nucleotide level. Here, we defined the genetic stability of murine cytomegalovirus (MCMV) by whole genome sequencing. Independently assembled sequences of three sister plaques showed only two single-base-pair substitutions after in vitro passage. In vivo passaged MCMV likewise demonstrated low mutation rates, comparable to those after in vitro passage, indicating high genome stability of MCMV at the single nucleotide level in the absence of obvious selection pressure. PMID- 20015992 TI - The potent anti-HIV activity of CXCL12gamma correlates with efficient CXCR4 binding and internalization. AB - We previously demonstrated that the naturally occurring splice variant stromal cell-derived factor 1gamma/CXCL12gamma is the most potent CXCL12 isoform in blocking X4 HIV-1, with weak chemotactic activity. A conserved BBXB domain (B for basic and X for any residue) located in the N terminus ((24)KHLK(27)) is found in all six isoforms of CXCL12. To determine whether the potent antiviral activity of CXCL12gamma is due to the presence of the extra C-terminal BBXB domains, we mutated each domain individually as well as in combination. Although binding of CXCL12gamma to heparan sulfate proteoglycan (HSPG) was 10-fold higher than that observed with CXCL12alpha, the results did not demonstrate a direct correlation between HSPG binding and the potent antiviral activity. CXCL12gamma mutants lacking the conserved BBXB domain (designated gammaB1) showed increased binding to HSPG but reduced anti-HIV activity. In contrast, the mutants lacking the C terminal second and/or third BBXB domain but retaining the conserved domain (designated B2, B3, and B23) showed decreased binding to HSPG but increased anti HIV activity. The B2, B3, and B23 mutants were associated with enhanced CXCR4 binding, receptor internalization, and restored chemotaxis. Internalization of CXCR4 was more potent with CXCL12gamma than with CXCL12alpha and was significantly reduced when the conserved BBXB domain was mutated. We concluded that the observed potent anti-HIV-1 activity of CXCL12gamma is due to increased affinity for CXCR4 and to efficient receptor internalization. PMID- 20015994 TI - Proteomic analysis of interactions between a deep-sea thermophilic bacteriophage and its host at high temperature. AB - The virus-host interaction is essential to understanding the role that viruses play in ecological and geochemical processes in deep-sea vent ecosystems. Virus induced changes in cellular gene expression and host physiology have been studied extensively. However, the molecular mechanism of interaction between a bacteriophage and its host at high temperature remains poorly understood. In the present study, the virus-induced gene expression profile of Geobacillus sp. E263, a thermophile isolated from a deep-sea hydrothermal ecosystem, was characterized. Based on proteomic analysis and random arbitrarily primed PCR (RAP-PCR) of Geobacillus sp. E263 cultured under non-bacteriophage GVE2 infection and GVE2 infection conditions, there were two types of protein/gene profiles in response to GVE2 infection. Twenty differentially expressed genes and proteins were revealed that could be grouped into 3 different categories based on cellular function, suggesting a coordinated response to infection. These differentially expressed genes and proteins were further confirmed by Northern blot analysis. To characterize the host proteins in response to virus infection, aspartate aminotransferase (AST) was inactivated to construct the AST mutant of Geobacillus sp. E263. The results showed that the AST protein was essential in virus infection. Thus, transcriptional and proteomic analyses and functional analysis revealed previously unknown host responses to deep-sea thermophilic virus infection. PMID- 20015995 TI - HIV-1 Nef inhibits ruffles, induces filopodia, and modulates migration of infected lymphocytes. AB - The HIV-1 Nef protein is a pathogenic factor modulating the behavior of infected cells. Nef induces actin cytoskeleton changes and impairs cell migration toward chemokines. We further characterized the morphology, cytoskeleton dynamics, and motility of HIV-1-infected lymphocytes. By using scanning electron microscopy, confocal immunofluorescence microscopy, and ImageStream technology, which combines flow cytometry and automated imaging, we report that HIV-1 induces a characteristic remodeling of the actin cytoskeleton. In infected lymphocytes, ruffle formation is inhibited, whereas long, thin filopodium-like protrusions are induced. Cells infected with HIV with nef deleted display a normal phenotype, and Nef expression alone, in the absence of other viral proteins, induces morphological changes. We also used an innovative imaging system to immobilize and visualize living individual cells in suspension. When combined with confocal "axial tomography," this technique greatly enhances three-dimensional optical resolution. With this technique, we confirmed the induction of long filopodium like structures in unfixed Nef-expressing lymphocytes. The cytoskeleton reorganization induced by Nef is associated with an important impairment of cell movements. The adhesion and spreading of infected cells to fibronectin, their spontaneous motility, and their migration toward chemokines (CXCL12, CCL3, and CCL19) were all significantly decreased. Therefore, Nef induces complex effects on the lymphocyte actin cytoskeleton and cellular morphology, which likely impacts the capacity of infected cells to circulate and to encounter and communicate with bystander cells. PMID- 20015996 TI - Targeted delivery of small interfering RNA to human dendritic cells to suppress dengue virus infection and associated proinflammatory cytokine production. AB - Dengue is a common arthropod-borne flaviviral infection in the tropics, for which there is no vaccine or specific antiviral drug. The infection is often associated with serious complications such as dengue hemorrhagic fever (DHF) or dengue shock syndrome (DSS), in which both viral and host factors have been implicated. RNA interference (RNAi) is a potent antiviral strategy and a potential therapeutic option for dengue if a feasible strategy can be developed for delivery of small interfering RNA (siRNA) to dendritic cells (DCs) and macrophages, the major in vivo targets of the virus and also the source of proinflammatory cytokines. Here we show that a dendritic cell-targeting 12-mer peptide (DC3) fused to nona-D arginine (9dR) residues (DC3-9dR) delivers siRNA and knocks down endogenous gene expression in heterogenous DC subsets, (monocyte-derived DCs [MDDCs], CD34(+) hematopoietic stem cell [HSC])-derived Langerhans DCs, and peripheral blood DCs). Moreover, DC3-9dR-mediated delivery of siRNA targeting a highly conserved sequence in the dengue virus envelope gene (siFvE(D)) effectively suppressed dengue virus replication in MDDCs and macrophages. In addition, DC-specific delivery of siRNA targeting the acute-phase cytokine tumor necrosis factor alpha (TNF-alpha), which plays a major role in dengue pathogenesis, either alone or in combination with an antiviral siRNA, significantly reduced virus-induced production of the cytokine in MDDCs. Finally to validate the strategy in vivo, we tested the ability of the peptide to target human DCs in the NOD/SCID/IL-2Rgamma( /-) mouse model engrafted with human CD34(+) hematopoietic stem cells (HuHSC mice). Treatment of mice by intravenous (i.v.) injection of DC3-9dR-complexed siRNA targeting TNF-alpha effectively suppressed poly(I:C)-induced TNF-alpha production by DCs. Thus, DC3-9dR can deliver siRNA to DCs both in vitro and in vivo, and this delivery approach holds promise as a therapeutic strategy to simultaneously suppress virus replication and curb virus-induced detrimental host immune responses in dengue infection. PMID- 20015997 TI - Role of untranslated regions in regulation of gene expression, replication, and pathogenicity of Newcastle disease virus expressing green fluorescent protein. AB - To gain insight into the role of untranslated regions (UTRs) in regulation of foreign gene expression, replication, and pathogenicity of Newcastle disease virus (NDV), a green fluorescent protein (GFP) gene flanked by 5' and 3' UTRs of each NDV gene was individually expressed by recombinant NDVs. UTRs of each gene modulated GFP expression positively or negatively. In particular, UTRs of the M and F genes enhanced levels of GFP expression at the junction of the P and M genes without altering replication of NDV, suggesting that UTRs could be used for enhanced expression of a foreign gene by NDV. PMID- 20015998 TI - Genetic and pathobiologic characterization of pandemic H1N1 2009 influenza viruses from a naturally infected swine herd. AB - Since its initial identification in Mexico and the United States, concerns have been raised that the novel H1N1 influenza virus might cause a pandemic of severity comparable to that of the 1918 pandemic. In late April 2009, viruses phylogenetically related to pandemic H1N1 influenza virus were isolated from an outbreak on a Canadian pig farm. This outbreak also had epidemiological links to a suspected human case. Experimental infections carried out in pigs using one of the swine isolates from this outbreak and the human isolate A/Mexico/InDRE4487/2009 showed differences in virus recovery from the lower respiratory tract. Virus was consistently isolated from the lungs of pigs infected with A/Mexico/InDRE4487/2009, while only one pig infected with A/swine/Alberta/OTH-33-8/2008 yielded live virus from the lung, despite comparable amounts of viral RNA and antigen in both groups of pigs. Clinical disease resembled other influenza virus infections in swine, albeit with somewhat prolonged virus antigen detection and delayed viral-RNA clearance from the lungs. There was also a noteworthy amount of genotypic variability among the viruses isolated from the pigs on the farm. This, along with the somewhat irregular pathobiological characteristics observed in experimentally infected animals, suggests that although the virus may be of swine origin, significant viral evolution may still be ongoing. PMID- 20015999 TI - Genomewide association analysis of respiratory syncytial virus infection in mice. AB - Respiratory syncytial virus (RSV) is the major cause of lower respiratory tract infection in infants, with about half being infected in their first year of life. Yet only 2 to 3% of infants are hospitalized for RSV infection, suggesting that individual susceptibility contributes to disease severity. Previously, we determined that AKR/J (susceptible) mice developed high lung RSV titers and showed delayed weight recovery, whereas C57BL/6J (resistant) mice demonstrated low lung RSV titers and rapid weight recovery. In addition, we have reported that gene-targeted mice lacking the cystic fibrosis transmembrane conductance regulator (Cftr; ATP-binding cassette subfamily C, member 7) are susceptible to RSV infection. For this report, recombinant backcross and F2 progeny derived from C57BL/6J and AKR/J mice were infected with RSV, their lung titers were measured, and quantitative trait locus (QTL) analysis was performed. A major QTL, designated Rsvs1, was identified on proximal mouse chromosome 6 in both recombinant populations. Microarray analysis comparing lung transcripts of the parental strains during infection identified several candidate genes that mapped to the Rsvs1 interval, including Cftr. These findings add to our understanding of individual RSV susceptibility and strongly support a modifier role for CFTR in RSV infection, a significant cause of respiratory morbidity in infants with cystic fibrosis. PMID- 20016000 TI - Activation of the N-terminally truncated form of the Stk receptor tyrosine kinase Sf-Stk by Friend virus-encoded gp55 is mediated by cysteine residues in the ecotropic domain of gp55 and the extracellular domain of Sf-Stk. AB - Friend virus induces an erythroleukemia in susceptible mice that is initiated by the interaction of the Friend virus-encoded glycoprotein gp55 with the erythropoietin (Epo) receptor and the product of the host Fv2 gene, a naturally occurring truncated form of the Stk receptor tyrosine kinase (Sf-Stk). We have previously demonstrated that the activation of Sf-Stk, recruitment of a Grb2/Gab2/Stat3 signaling complex, and induction of Pu.1 expression by Stat3 are required for the development of the early stage of Friend disease both in vitro and in vivo. Here we demonstrate that the interaction of gp55 with Sf-Stk is dependent on cysteine residues in the ecotropic domain of gp55 and the extracellular domain of Sf-Stk. Point mutation of these cysteine residues or deletion of these domains inhibits the ability of gp55 to interact with Sf-Stk, resulting in the inability of these proteins to promote the Epo-independent growth of erythroid progenitor cells. We also demonstrate that the interaction of gp55 with Sf-Stk does not promote dimerization of Sf-Stk but results in enhanced phosphorylation of Sf-Stk and the relocalization of Sf-Stk from the cytosol to the plasma membrane. Finally, we demonstrate that a constitutively active form of Sf-Stk (Sf-StkM330T), as well as its human counterpart, Sf-Ron, promotes Epo independent colony formation in the absence of gp55 and that this response is also dependent on the cysteines in the extracellular domains of Sf-StkM330T and Sf-Ron. These data suggest that the cysteines in the extracellular domains of Sf Stk and Sf-Ron may also mediate the interaction of these truncated receptors with other cellular factors that regulate their ability to promote cytokine independent growth. PMID- 20016001 TI - Connectivity analysis reveals a cortical network for eye gaze perception. AB - Haxby et al. (Haxby JV, Hoffman EA, Gobbini MI. 2000. The distributed human neural system for face perception. Trends Cogn Sci. 4:223-233.) proposed that eye gaze processing results from an interaction between a "core" face-specific system involved in visual analysis and an "extended" system involved in spatial attention, more generally. However, the full gaze perception network has remained poorly specified. In the context of a functional magnetic resonance imaging study, we used psychophysiological interactions (PPIs) to identify brain regions that showed differential connectivity (correlation) with core face perception structures (posterior superior temporal sulcus [pSTS] and fusiform gyrus [FG]) when viewing gaze shifts relative to control eye movements (opening/closing the eyes). The PPIs identified altered connectivity between the pSTS and MT/V5, intraparietal sulcus, frontal eye fields, superior temporal gyrus (STG), supramarginal gyrus, and middle frontal gyrus (MFG). The FG showed altered connectivity with the same areas of the STG and MFG, demonstrating the contribution of both dorsal and ventral core face areas to gaze perception. We propose that this network provides an interactive system that alerts us to seen changes in other agents' gaze direction, makes us aware of their altered focus of spatial attention, and prepares a corresponding shift in our own attention. PMID- 20016002 TI - Mkl transcription cofactors regulate structural plasticity in hippocampal neurons. AB - Expressed throughout the central nervous system, the myocardin-related, megakaryoblastic acute leukemia 1 and 2 (Mkl1/2) are transcriptional cofactors that can be found tethered in the cytoplasm to monomeric actin but on synaptic activation translocate to the nucleus and associate with transcription factors such as serum response factor (SRF) to regulate expression of structural genes. This implies a potential role for Mkls in linking synaptic activity, through gene expression control, to neuronal structural plasticity. Here, we present evidence that Mkls, particularly Mkl2, are powerful regulators of neuronal structure in vitro. Moreover, using the passive avoidance-conditioning paradigm, we identify learning-associated alterations of neuronal Mkl expression that appear to contribute to 2 phases of gene regulation during memory consolidation in the hippocampus. Gene regulation immediately after learning includes Egr2 and may be facilitated by downregulation of Mkls likely releasing ternary complex factor regulated SRF activity. The second transcriptional phase occurs later at the 3-h postavoidance time point when Mkl accumulates in the nucleus of hippocampal neurons and there is enhanced transcription of Mkl-dependent structural genes that may contribute to the elaboration of new, memory-associated synapses known to appear over the subsequent 3-h period. PMID- 20016003 TI - Semantic, factual, and social language comprehension in adolescents with autism: an FMRI study. AB - Language in high-functioning autism is characterized by pragmatic and semantic deficits, and people with autism have a reduced tendency to integrate information. Because the left and right inferior frontal (LIF and RIF) regions are implicated with integration of speaker information, world knowledge, and semantic knowledge, we hypothesized that abnormal functioning of the LIF and RIF regions might contribute to pragmatic and semantic language deficits in autism. Brain activation of sixteen 12- to 18-year-old, high-functioning autistic participants was measured with functional magnetic resonance imaging during sentence comprehension and compared with that of twenty-six matched controls. The content of the pragmatic sentence was congruent or incongruent with respect to the speaker characteristics (male/female, child/adult, and upper class/lower class). The semantic- and world-knowledge sentences were congruent or incongruent with respect to semantic expectancies and factual expectancies about the world, respectively. In the semantic-knowledge and world-knowledge condition, activation of the LIF region did not differ between groups. In sentences that required integration of speaker information, the autism group showed abnormally reduced activation of the LIF region. The results suggest that people with autism may recruit the LIF region in a different manner in tasks that demand integration of social information. PMID- 20016004 TI - Janus-faced enzymes yeast Tgl3p and Tgl5p catalyze lipase and acyltransferase reactions. AB - In the yeast, mobilization of triacylglycerols (TAGs) is facilitated by the three TAG lipases Tgl3p, Tgl4p, and Tgl5p. Motif search analysis, however, indicated that Tgl3p and Tgl5p do not only contain the TAG lipase motif GXSXG but also an H (X)(4)-D acyltransferase motif. Interestingly, lipid analysis revealed that deletion of TGL3 resulted in a decrease and overexpression of TGL3 in an increase of glycerophospholipids. Similar results were obtained with TGL5. Therefore, we tested purified Tgl3p and Tgl5p for acyltransferase activity. Indeed, both enzymes not only exhibited lipase activity but also catalyzed acylation of lysophosphatidylethanolamine and lysophosphatidic acid, respectively. Experiments using variants of Tgl3p created by site-directed mutagenesis clearly demonstrated that the two enzymatic activities act independently of each other. We also showed that Tgl3p is important for efficient sporulation of yeast cells, but rather through its acyltransferase than lipase activity. In summary, our results demonstrate that yeast Tgl3p and Tgl5p play a dual role in lipid metabolism contributing to both anabolic and catabolic processes. PMID- 20016005 TI - Compartment-specific synthesis of phosphatidylethanolamine is required for normal heavy metal resistance. AB - Control of lipid composition of membranes is crucial to ensure normal cellular functions. Saccharomyces cerevisiae has two different phosphatidylserine decarboxylase enzymes (Psd1 and Psd2) that catalyze formation of phosphatidylethanolamine. The mitochondrial Psd1 provides roughly 70% of the phosphatidylethanolamine (PE) biosynthesis in the cell with Psd2 carrying out the remainder. Here, we demonstrate that loss of Psd2 causes cells to acquire sensitivity to cadmium even though Psd1 remains intact. This cadmium sensitivity results from loss of normal activity of a vacuolar ATP-binding cassette transporter protein called Ycf1. Measurement of phospholipid levels indicates that loss of Psd2 causes a specific reduction in vacuolar membrane PE levels, whereas total PE levels are not significantly affected. The presence of a phosphatidylinositol transfer protein called Pdr17 is required for Psd2 function and normal cadmium tolerance. We demonstrate that Pdr17 and Psd2 form a complex in vivo that seems essential for maintenance of vacuolar PE levels. Finally, we refine the localization of Psd2 to the endosome arguing that this enzyme controls vacuolar membrane phospholipid content by regulating phospholipids in compartments that will eventually give rise to the vacuole. Disturbance of this regulation of intracellular phospholipid balance leads to selective loss of membrane protein function in the vacuole. PMID- 20016006 TI - The beta subunit of the heterotrimeric G protein triggers the Kluyveromyces lactis pheromone response pathway in the absence of the gamma subunit. AB - The Kluyveromyces lactis heterotrimeric G protein is a canonical Galphabetagamma complex; however, in contrast to Saccharomyces cerevisiae, where the Ggamma subunit is essential for mating, disruption of the KlGgamma gene yielded cells with almost intact mating capacity. Expression of a nonfarnesylated Ggamma, which behaves as a dominant-negative in S. cerevisiae, did not affect mating in wild type and DeltaGgamma cells of K. lactis. In contrast to the moderate sterility shown by the single DeltaKlGalpha, the double DeltaKlGalpha DeltaKlGgamma mutant displayed full sterility. A partial sterile phenotype of the DeltaKlGgamma mutant was obtained in conditions where the KlGbeta subunit interacted defectively with the Galpha subunit. The addition of a CCAAX motif to the C-end of KlGbeta, partially suppressed the lack of both KlGalpha and KlGgamma subunits. In cells lacking KlGgamma, the KlGbeta subunit cofractionated with KlGalpha in the plasma membrane, but in the DeltaKlGalpha DeltaKlGgamma strain was located in the cytosol. When the KlGbeta-KlGalpha interaction was affected in the DeltaKlGgamma mutant, most KlGbeta fractionated to the cytosol. In contrast to the generic model of G-protein function, the Gbeta subunit of K. lactis has the capacity to attach to the membrane and to activate mating effectors in absence of the Ggamma subunit. PMID- 20016007 TI - Live-cell imaging in Caenorhabditis elegans reveals the distinct roles of dynamin self-assembly and guanosine triphosphate hydrolysis in the removal of apoptotic cells. AB - Dynamins are large GTPases that oligomerize along membranes. Dynamin's membrane fission activity is believed to underlie many of its physiological functions in membrane trafficking. Previously, we reported that DYN-1 (Caenorhabditis elegans dynamin) drove the engulfment and degradation of apoptotic cells through promoting the recruitment and fusion of intracellular vesicles to phagocytic cups and phagosomes, an activity distinct from dynamin's well-known membrane fission activity. Here, we have detected the oligomerization of DYN-1 in living C. elegans embryos and identified DYN-1 mutations that abolish DYN-1's oligomerization or GTPase activities. Specifically, abolishing self-assembly destroys DYN-1's association with the surfaces of extending pseudopods and maturing phagosomes, whereas inactivating guanosine triphosphate (GTP) binding blocks the dissociation of DYN-1 from these membranes. Abolishing the self assembly or GTPase activities of DYN-1 leads to common as well as differential phagosomal maturation defects. Whereas both types of mutations cause delays in the transient enrichment of the RAB-5 GTPase to phagosomal surfaces, only the self-assembly mutation but not GTP binding mutation causes failure in recruiting the RAB-7 GTPase to phagosomal surfaces. We propose that during cell corpse removal, dynamin's self-assembly and GTP hydrolysis activities establish a precise dynamic control of DYN-1's transient association to its target membranes and that this control mechanism underlies the dynamic recruitment of downstream effectors to target membranes. PMID- 20016008 TI - Two isoforms of Npap60 (Nup50) differentially regulate nuclear protein import. AB - Npap60 (Nup50) is a nucleoporin that binds directly to importin alpha. In humans, there are two Npap60 isoforms: the long (Npap60L) and short (Npap60S) forms. In this study, we provide both in vitro and in vivo evidence that Npap60L and Npap60S function differently in nuclear protein import. In vitro binding assays revealed that Npap60S stabilizes the binding of importin alpha to classical NLS cargo, whereas Npap60L promotes the release of NLS-cargo from importin alpha. In vivo time-lapse experiments showed that when the Npap60 protein level is controlled, allowing CAS to efficiently promote the dissociation of the Npap60/importin alpha complex, Npap60S and Npap60L suppress and accelerate the nuclear import of NLS-cargo, respectively. These results demonstrate that Npap60L and Npap60S have opposing functions and suggest that Npap60L and Npap60S levels must be carefully controlled for efficient nuclear import of classical NLS-cargo in humans. This study provides novel evidence that nucleoporin expression levels regulate nuclear import efficiency. PMID- 20016009 TI - GRASP and IPCEF promote ARF-to-Rac signaling and cell migration by coordinating the association of ARNO/cytohesin 2 with Dock180. AB - ARFs are small GTPases that regulate vesicular trafficking, cell shape, and movement. ARFs are subject to extensive regulation by a large number of accessory proteins. The many different accessory proteins are likely specialized to regulate ARF signaling during particular processes. ARNO/cytohesin 2 is an ARF activating protein that promotes cell migration and cell shape changes. We report here that protein-protein interactions mediated by the coiled-coil domain of ARNO are required for ARNO induced motility. ARNO lacking the coiled-coil domain does not promote migration and does not induce ARF-dependent Rac activation. We find that the coiled-coil domain promotes the assembly of a multiprotein complex containing both ARNO and the Rac-activating protein Dock180. Knockdown of either GRASP/Tamalin or IPCEF, two proteins known to bind to the coiled-coil of ARNO, prevents the association of ARNO and Dock180 and prevents ARNO-induced Rac activation. These data suggest that scaffold proteins can regulate ARF dependent processes by biasing ARF signaling toward particular outputs. PMID- 20016011 TI - Effectiveness of a large-scale iron-fortified milk distribution program on anemia and iron deficiency in low-income young children in Mexico. AB - BACKGROUND: Anemia during infancy impairs neurodevelopment. Little information has been published about the effectiveness of large-scale programs on anemia and iron-deficiency prevention. OBJECTIVE: The objective was to assess the effectiveness of a large-scale program that distributes subsidized iron-fortified milk in Mexico on anemia and iron deficiency in children aged 12-30 mo. DESIGN: A double-blinded, group-randomized effectiveness trial was conducted in 12 milk distribution clusters assigned to consume iron-fortified (FM; n = 7) or nonfortified (NFM; n = 5) milk. A daily portion of FM contained 5.28 mg Fe (ferrous gluconate) and 48 mg sodium ascorbate. RESULTS: Overall treatment effects were documented at 6 and 12 mo for anemia and for iron deficiency assessed by both serum ferritin (SF) and serum soluble transferrin receptor (sTfR) (interaction: P < 0.10). Differential effects at 6 mo (P = 0.004) and 12 mo (P = 0.664) were documented only for sTfR. Estimated prevalences (EPs) of anemia (hemoglobin < 110 g/L) from baseline to 6 and 12 mo decreased from 42.6% to 19.7% and 9.4%, respectively, in the NFM group (n = 210) and from 44.5% to 12.7% and 4.0%, respectively, in the FM group (n = 357). EPs of SF < 12 mug/L from baseline to 6 and 12 mo changed from 36.0% to 41.8% and 17.1%, respectively, in the NFM group (n = 43) and from 29.8% to 18.6% and 5.7%, respectively, in the FM group (n = 144). EPs of sTfR > 3.3 mg/L from baseline to 6 and 12 mo decreased from 16.2% to 8.3% and 2.0%, respectively, in the NFM group (n = 114) and from 15.5% to 0.7% and 1.1%, respectively, in the FM group. CONCLUSION: A large-scale iron-fortified subsidized-milk program was effective at reducing the rates of anemia and iron deficiency in Mexican children during 12 mo of implementation. This trial was registered at clinicaltrials.gov as NCT00508131. PMID- 20016010 TI - Consumption of sodium and salted foods in relation to cancer and cardiovascular disease: the Japan Public Health Center-based Prospective Study. AB - BACKGROUND: Although the influence of salt, per se, on the risk of cancer or cardiovascular disease (CVD) might differ from that of salt-preserved foods, few studies have simultaneously examined the effects of sodium and salted foods on the risk of either cancer or CVD. OBJECTIVE: We simultaneously examined associations between sodium and salted food consumption and the risk of cancer and CVD. DESIGN: During 1995-1998, a validated food-frequency questionnaire was administered to 77,500 men and women aged 45-74 y. During up to 598,763 person years of follow-up until the end of 2004, 4476 cases of cancer and 2066 cases of CVD were identified. RESULTS: Higher consumption of sodium was associated with a higher risk of CVD but not with the risk of total cancer: multivariate hazard ratios for the highest compared with lowest quintiles of intake were 1.19 (95% CI: 1.01, 1.40; P for trend: 0.06) for CVD and 1.04 (95% CI: 0.93, 1.16; P for trend: 0.63) for total cancer. Higher consumption of salted fish roe was associated with higher risk of total cancer, and higher consumption of cooking and table salt was associated with higher risk of CVD. Similar results were seen for the risk of gastric or colorectal cancer and stroke. CONCLUSIONS: Sodium intake as a whole salt equivalent may not increase the risk of cancer but may increase that of CVD. In contrast, salted food intake may increase the risk of cancer. Our findings support the notion that sodium and salted foods have differential influences on the development of cancer and CVD. PMID- 20016012 TI - Effects of daily snack food intake on food reinforcement depend on body mass index and energy density. AB - BACKGROUND: The reinforcing value of food plays a role in food consumption. We have shown previously that daily intake of a high-energy-density (HED) snack food decreases food reinforcement and food liking in nonobese women but increases food reinforcement and decreases food liking in obese women. OBJECTIVE: These previous studies were conducted with the use of only HED snack foods. The purpose of this study was to determine whether these effects generalize to low-energy-density (LED) foods. DESIGN: Participants (n = 53) had food reinforcement and food liking tested at baseline and then again after 2 wk of daily consumption of 60-g portions of an HED (n = 26) or an LED (n = 27) snack food. RESULTS: We observed a decrease in food reinforcement in women with a lower body mass index (BMI) and an increase in food reinforcement in women with a higher BMI after 14 d of consumption of an HED snack food. Food liking decreased in all women, regardless of BMI, after repeated consumption of HED foods. Conversely, all women, regardless of BMI, showed a decrease in food reinforcement after 14 d of LED snack food consumption. Women with a lower BMI who consumed LED snacks also showed a decrease in liking, but women with a higher BMI who consumed LED foods reported no change in liking. CONCLUSIONS: These findings suggest that changes in food reinforcement after daily snack food intake are influenced by both BMI and the energy density of the foods. In addition, changes in food reinforcement cannot be explained by changes in food liking. PMID- 20016013 TI - Nonheme-iron absorption in first-degree relatives is highly correlated: a stable isotope study in mother-child pairs. AB - BACKGROUND: Iron absorption in humans is highly variable even after iron status and dietary components that influence iron absorption are controlled for. Inherited factors may help explain this variance. OBJECTIVE: Our objective was to compare nonheme-iron absorption from a noninhibitory, stable-isotope-labeled test meal in preschool-aged children and their mothers. DESIGN: We provided 72 test meals based on degermed maize flour and milk powder and fortified with [(57)Fe]ferrous fumarate or [(58)Fe]ferrous sulfate to healthy Mexican preschool children [n = 18; mean (+/-SD) age: 3.6 +/- 1.0 y] and their mothers [n = 18; mean (+/-SD) age: 28.0 +/- 5.2 y]. Iron absorption was calculated on the basis of incorporation of isotopes into erythrocytes after 14 d and was adjusted for differences in iron status. RESULTS: There was a wide variation in iron absorption from the test meals: in the mothers and children, the median fractional absorption of ferrous sulfate was 22.55% (range: 1.65-54.83%) and 5.51% (range: 2.23-17.20%), respectively (P < 0.0001). After adjustment for serum ferritin, the significant difference in absorption between mothers and their children disappeared. Despite this broad range of iron absorption, corrected fractional iron absorption from the ferrous fumarate-fortified (r(2) = 0.582) and the ferrous sulfate-fortified test meals (r(2) = 0.557) was strongly correlated in mothers and their children (P < 0.0001). There was a striking positive correlation between the mean corrected fractional iron absorption from both test meals in mothers and their children (r(2) = 0.782, P < 0.0001). In regression analyses that included age, sex, and hemoglobin, the only significant predictor of corrected fractional iron absorption in children was corrected fractional iron absorption in their mothers (standardized beta = 0.884, P < 0.001). CONCLUSIONS: Nonheme-iron absorption exhibits a strong familial tendency. After differences in meal matrix and serum ferritin are accounted for, these data suggest that inheritance and/or shared environmental factors explain most of the variance in dietary iron absorption. PMID- 20016014 TI - Consumption of restaurant foods and incidence of type 2 diabetes in African American women. AB - BACKGROUND: Type 2 diabetes is a major problem in Western nations. Profound secular changes in the food environment and eating habits may play a role. In particular, consumption of foods prepared outside the home has greatly increased. OBJECTIVE: We investigated the relation of restaurant meal consumption to incidence of type 2 diabetes among African American women with the use of data from the prospective Black Women's Health Study. DESIGN: The participants have completed mailed follow-up questionnaires every 2 y since 1995, including food frequency questionnaires that asked about the frequency of eating restaurant meals of various types. Cox proportional hazards models were used to calculate incidence rate ratios and 95% CIs for the association of type 2 diabetes incidence with various categories of consumption of each restaurant food relative to the lowest category, with adjustment for diabetes risk factors. RESULTS: Among 44,072 participants aged 30-69 y and free of diabetes at baseline, 2873 incident cases of type 2 diabetes occurred during 10 y of follow-up. Consumption of restaurant meals of hamburgers, fried chicken, fried fish, and Chinese food were independently associated with an increased risk of type 2 diabetes. Incidence rate ratios for > or = 2 such meals per week relative to none were 1.40 (95% CI: 1.14, 1.73) for hamburgers and 1.68 (95% CI: 1.36, 2.08) for fried chicken. Control for body mass index greatly reduced the estimates, which suggests that the associations are mediated through weight gain and obesity. CONCLUSION: The present study has identified a risk factor for type 2 diabetes that may be readily modifiable by dietary changes. PMID- 20016015 TI - Dietary fiber and subsequent changes in body weight and waist circumference in European men and women. AB - BACKGROUND: Dietary fiber may play a role in obesity prevention. Until now, the role that fiber from different sources plays in weight change had rarely been studied. OBJECTIVE: Our aim was to investigate the association of total dietary fiber, cereal fiber, and fruit and vegetable fiber with changes in weight and waist circumference. DESIGN: We conducted a prospective cohort study with 89,432 European participants, aged 20-78 y, who were free of cancer, cardiovascular disease, and diabetes at baseline and who were followed for an average of 6.5 y. Dietary information was collected by using validated country-specific food frequency questionnaires. Multiple linear regression analysis was performed in each center studied, and estimates were combined by using random-effects meta analyses. Adjustments were made for follow-up duration, other dietary variables, and baseline anthropometric, demographic, and lifestyle factors. RESULTS: Total fiber was inversely associated with subsequent weight and waist circumference change. For a 10-g/d higher total fiber intake, the pooled estimate was -39 g/y (95% CI: -71, -7 g/y) for weight change and -0.08 cm/y (95% CI: -0.11, -0.05 cm/y) for waist circumference change. A 10-g/d higher fiber intake from cereals was associated with -77 g/y (95% CI: -127, -26 g/y) weight change and -0.10 cm/y (95% CI: -0.18, -0.02 cm/y) waist circumference change. Fruit and vegetable fiber was not associated with weight change but had a similar association with waist circumference change when compared with intake of total dietary fiber and cereal fiber. CONCLUSION: Our finding may support a beneficial role of higher intake of dietary fiber, especially cereal fiber, in prevention of body-weight and waist circumference gain. PMID- 20016016 TI - How many struggle to get by in retirement? AB - PURPOSE: This study compared the official poverty rate for adults aged 65 years and older with alternative measures that portray the true resources and needs of older adults. DESIGN AND METHODS: The analysis used data from the 2004 Health and Retirement Study on income, assets, in-kind transfers, and out-of-pocket medical expenses. It also incorporated the effects of federal and state income taxes on net incomes, and accounted for the potential annuity value of assets in measuring resources. RESULTS: The alternative poverty measures that account for out-of pocket health spending estimate that between 0.3 and 1.5 million more older adults live in poverty than acknowledged by the official measure. IMPLICATIONS: These results highlight the need to update poverty measures so that they better reflect older adults' circumstances. This will become increasingly important as policymakers consider reforms to improve solvency in the Social Security and Medicare programs that may require benefit cuts or increased cost sharing. PMID- 20016017 TI - New therapeutic strategies for raltegravir. AB - Raltegravir (Isentress) is the first approved HIV integrase inhibitor. Agents in this class target a different viral enzyme compared with agents inhibiting reverse transcriptase and protease. A wide number of patients are currently susceptible to integrase inhibitors, including heavily antiretroviral-experienced patients harbouring drug-resistant viruses. The good tolerability and convenience of raltegravir have recently begun to be appreciated, leading to the consideration of other indications for the drug. Data recently released using the drug as first-line therapy or in switch strategies are very promising and the role of raltegravir in intensification therapy is currently under investigation. Altogether, the current information supports a broad use of raltegravir beyond its initial approval for antiretroviral-experienced HIV-infected patients. PMID- 20016018 TI - Association between plasma levels of eotaxin (CCL-11) and treatment response to interferon-alpha and ribavirin in HIV/HCV co-infected patients. AB - OBJECTIVES: To analyse the association between plasma chemokine levels at baseline and virological response to interferon-alpha (IFN-alpha) + ribavirin in human immunodeficiency virus (HIV)/hepatitis C virus (HCV) co-infected patients. METHODS: We carried out a retrospective study in 109 patients. Chemokines were measured using Multiplex kits using a Luminex 100 Analyzer. Logistic regression was used to evaluate the association between plasma chemokine levels before HCV therapy and virological response at weeks 48 and 72 after starting HCV therapy. RESULTS: Fifty-seven patients out of 103 achieved end of treatment virological response (ETR). In patients achieving ETR, the baseline levels of eotaxin, MCP-1 and MCP-3 were higher than non-responder (NR) patients. Similarly, 51 patients out of 106 achieved sustained virological response (SVR). In patients achieving SVR, the baseline levels of eotaxin and MCP-1 were higher than in NR patients. Plasma levels of eotaxin, MCP-1 and MCP-3 had a significant positive association with ETR, as well as eotaxin and MCP-1 with SVR. However, after stepwise multivariate logistic regression, eotaxin was the only chemokine selected capable of predicting ETR and SVR with odds ratio (OR) of 1.016 (95% CI: 1.004-1.029) and 1.015 (95% CI: 1.002-1.027) for ETR and SVR, respectively. CONCLUSIONS: Our preliminary data suggest that plasma eotaxin levels prior to HCV antiviral therapy may be useful in predicting virological response to HCV treatment with IFN-alpha + ribavirin in HIV/HCV co-infected patients. Further experimental research is necessary to corroborate this hypothesis. PMID- 20016020 TI - Caspofungin-non-susceptible Candida isolates in cancer patients. AB - OBJECTIVES: To identify the frequency of caspofungin-non-susceptible Candida isolates in cancer patients with candidiasis. METHODS: We reviewed the in vitro susceptibilities (M27-A3 CLSI method) of 650 Candida spp. associated with invasive candidiasis episodes in 582 hospitalized cancer patients (2005-08). RESULTS AND CONCLUSIONS: We identified seven caspofungin-non-susceptible Candida strains (three Candida tropicalis, two Candida glabrata and two Candida albicans) from 650 Candida isolates (1%). C. tropicalis (three out of seven) was the most common non-susceptible species isolated. All patients responded to a change of antifungal therapy. Further surveillance should focus on the potential broader emergence of echinocandin resistance, as the clinical use of this antifungal class continues to expand in cancer patients. PMID- 20016021 TI - Ligand-induced regulation and localization of cannabinoid CB1 and dopamine D2L receptor heterodimers. AB - The cannabinoid CB(1) (CB(1)) and dopamine D(2) (D(2)) receptors are coexpressed in the basal ganglia, an area of the brain involved in such processes as cognition, motor function, and emotional control. Several lines of evidence suggest that CB(1) and D(2) receptors may oligomerize, providing a unique pharmacology in vitro and in vivo. However, limited information exists on the regulation of CB(1) and D(2) receptor dimers. We used a novel technique, multicolor bimolecular fluorescence complementation (MBiFC) to examine the subcellular localization of CB(1)-D(2L) heterodimers as well as D(2L)-D(2L) homodimers in a neuronal cell model, Cath. a differentiated cells. MBiFC was then used to explore the effects of persistent ligand treatment on receptor dimerization at the plasma membrane and intracellularly. Persistent (20-h) agonist treatment resulted in increased formation of CB(1)-D(2L) heterodimers relative to the D(2L)-D(2L) homodimers. The effects of the D(2) agonist quinpirole were restricted to the intracellular compartment and may reflect increased D(2L) receptor expression. In contrast, treatment with the CB(1) receptor agonist (2)-cis-3-[2-hydroxy-4-(1,1-dimethylheptyl)phenyl]-trans-4-(3 hydroxypropyl) cyclohexanol (CP55, 940) produced increases in both membrane and intracellular CB(1)-D(2L) heterodimers independently of alterations in CB(1) receptor expression. The effects of CB(1) receptor activation were attenuated by the CB(1) antagonist 1-(2,4-dichlorophenyl)-5-(4-iodophenyl)-4-methyl-N-4 morpholinyl-1H-pyrazole-3-carboxamide (AM281) and were both time- and dose dependent. The effects of CB(1) activation were examined further by combining MBiFC with a constitutively active CB(1) receptor mutant, CB(1)T210I. These studies demonstrated that the expression of CB(1)T210I increased intracellular CB(1)-D(2L) heterodimer formation. In summary, agonist-induced modulation of CB(1)-D(2L) oligomerization may have physiological implications in diseases such as Parkinson's disease and drug abuse. PMID- 20016023 TI - Editor's choice: citation classics. PMID- 20016022 TI - Hepatic mitochondrial DNA depletion after an alcohol binge in mice: probable role of peroxynitrite and modulation by manganese superoxide dismutase. AB - Alcohol consumption increases reactive oxygen species (ROS) formation, which can damage mitochondrial DNA (mtDNA) and alter mitochondrial function. To test whether manganese superoxide dismutase (MnSOD) modulates acute alcohol-induced mitochondrial alterations, transgenic MnSOD-overexpressing (MnSOD(+++)) mice, heterozygous knockout (MnSOD(+/-)) mice, and wild-type (WT) littermates were sacrificed 2 or 24 h after intragastric ethanol administration (5 g/kg). Alcohol administration further increased MnSOD activity in MnSOD(+++) mice, but further decreased it in MnSOD(+/-) mice. In WT mice, alcohol administration transiently increased mitochondrial ROS formation, decreased mitochondrial glutathione, depleted and damaged mtDNA, and decreased complex I and V activities; alcohol durably increased inducible nitric-oxide synthase (NOS) expression, plasma nitrites/nitrates, and the nitration of tyrosine residues in complex V proteins. These effects were prevented in MnSOD(+++) mice and prolonged in MnSOD(+/-) mice. In alcoholized WT or MnSOD(+/-) mice, mtDNA depletion and the nitration of tyrosine residues in complex I and V proteins were prevented or attenuated by cotreatment with tempol (4-hydroxy-2,2,6,6-tetramethylpiperidine-N-oxyl), a superoxide scavenger; N(omega)-nitro-l-arginine methyl ester and N-[3 (aminomethyl)benzyl]acetamidine (1,400W), two NOS inhibitors; or uric acid, a peroxynitrite scavenger. In conclusion, MnSOD overexpression prevents, and MnSOD deficiency prolongs, mtDNA depletion after an acute alcohol binge in mice. The protective effects of MnSOD, tempol, NOS inhibitors, and uric acid point out a role of the superoxide anion reacting with NO to form mtDNA-damaging peroxynitrite. PMID- 20016024 TI - Children born after preimplantation genetic diagnosis show no increase in congenital anomalies. PMID- 20016025 TI - Arcuate nucleus proopiomelanocortin neurons mediate the acute anorectic actions of leukemia inhibitory factor via gp130. AB - The proinflammatory cytokine leukemia inhibitory factor (LIF) is induced in disease states and is known to inhibit food intake when administered centrally. However, the neural pathways underlying this effect are not well understood. We demonstrate that LIF acutely inhibits food intake by directly activating pro opiomelanocortin (POMC) neurons in the arcuate nucleus of the hypothalamus. We show that arcuate POMC neurons express the LIF-R, and that LIF stimulates the release of the anorexigenic peptide, alpha-MSH from ex vivo hypothalami. Transgenic mice lacking gp130, the signal transducing subunit of the LIF-R complex, specifically in POMC neurons fail to respond to LIF. Furthermore, LIF does not stimulate the release of alpha-MSH from the transgenic hypothalamic explants. These findings indicate that POMC neurons mediate the acute anorectic actions of central LIF administration and provide a mechanistic link between inflammation and food intake. PMID- 20016026 TI - Nanoparticles and the immune system. AB - Today nanotechnology is finding growing applications in industry, biology, and medicine. The clear benefits of using nanosized products in various biological and medical applications are often challenged by concerns about the lack of adequate data regarding their toxicity. One area of interest involves the interactions between nanoparticles and the components of the immune system. Nanoparticles can be engineered to either avoid immune system recognition or specifically inhibit or enhance the immune responses. We review herein reported observations on nanoparticle-mediated immunostimulation and immunosuppression, focusing on possible theories regarding how manipulation of particle physicochemical properties can influence their interaction with immune cells to attain desirable immunomodulation and avoid undesirable immunotoxicity. PMID- 20016027 TI - Nanoparticles for molecular imaging--an overview. AB - Molecular imaging is a technique for quantifying physiological changes in vivo using imaging probes, or beacons, which can be detected noninvasively. This field of study has advanced rapidly in recent years, in part due to the application of nanotechnology. The versatility of different imaging modalities has been significantly enhanced by innovative nanoparticle development. These nanoprobes can be used to image specific cells and tissues within a whole organism. Some of the nanoparticles under development may be useful to measure biological processes associated with human disease and help monitor how these change with treatment. This review highlights some of the recent advances in nanoparticles for molecular imaging. It also addresses issues that arise with the use of nanoparticles. Whereas much of the technology remains at an experimental stage, the potential for enhancing disease diagnosis and treatment is considerable. PMID- 20016028 TI - A tiny touch: activation of cell signaling pathways with magnetic nanoparticles. AB - Magnetic nanoparticles can be coated with specific ligands that enable them to bind to receptors on a cell's surface. When a magnetic field is applied, it pulls on the particles so that they deliver nanoscale forces at the ligand-receptor bond. It has been observed that mechanical stimulation in this manner can activate cellular signaling pathways that are known as mechanotransduction pathways. Integrin receptors, stretch-activated ion channels, focal adhesions, and the cytoskeleton are key players in activating these pathways, but there is still much we do not know about how these mechanosensors work. Current evidence indicates that applied forces at these structures can activate Ca(2+) signaling, Src family protein kinase, MAPK, and RhoGTPase pathways. The techniques of magnetic twisting and magnetic tweezers, which use magnetic particles to apply forces to cells, afford a fine degree of control over how cells are stimulated and hold much promise in elucidating the fundamentals of mechanotransduction. The particles are generally not harmful to cellular health, and their nanoscale dimensions make them advantageous for probing a cell's molecular-scale sensory structures. This review highlights the basic aspects of magnetic nanoparticles, magnetic particle techniques and the structures and pathways that are involved in mechanotransduction. PMID- 20016029 TI - 7alpha-hydroxytestosterone affects 1 beta-hydroxysteroid dehydrogenase 1 direction in rat Leydig cells. AB - The cytochrome P450 2A1 (CYP2A1) is a P450 enzyme that catalyzes the metabolism of testosterone. CYP2A1 has been reported to be present in rat testis. However, its developmental changes and function have not been well characterized. The purpose of this study was to measure the abundance of CYP2A1 (Cyp2a1) mRNA in the developing rat testis and Leydig cells and examine the effects of its product, 7 alpha-hydroxytestosterone (7HT), on an important enzyme, 11 beta-hydroxysteroid dehydrogenase type 1 (11 beta-HSD1) that interconverts active corticosterone and inactive 11-dehydrocorticosterone. As detected by real-time PCR, Cyp2a1 was found to be present exclusively in the Leydig cell. CYP2A1 activity in adult Leydig cells was 5-fold higher than those in progenitor or immature Leydig cells. 7HT competitively suppressed 11 beta-HSD1 oxidase and reductase activities in rat testis microsome with inhibitory constant of 1.2 and 2.9 mum, respectively. In intact Leydig cells, 7HT did not inhibit 11 beta-HSD1 reductase activity, but it stimulated its reductase activity. Thus, at 100 nm and higher concentrations, 7HT significantly switched 11 beta-HSD1 oxidoreductase activities toward reductase. The present data shows that 7HT, the product formed by CYP2A1 from testosterone, regulates the direction of 11 beta-HSD1 activity in rat Leydig cells. PMID- 20016030 TI - Nutritional programming affects hypothalamic organization and early response to leptin. AB - Nutritional programming, taking place in utero or early after birth, is closely linked with metabolic and appetite disorders in adulthood. Following the hypothesis that nutritional programming impacts hypothalamic neuronal organization, we report on discrepancies of multiple molecular and cellular early events that take place in the hypothalamus of rats submitted to intrauterine growth restriction (IUGR). Expression screening performed on hypothalami from IUGR rats at birth and at postnatal d 12 identified changes in gene expression of neurodevelopmental process (cell differentiation and cytoskeleton organization). Additionally, a slight reduction of agouti-related protein and a strong reduction of alpha-MSH-immunoreactive efferent fibers were demonstrated in the paraventricular nucleus of IUGR rats. Rapid catch-up growth of IUGR rats, 5 d after birth, had a positive effect on neurodevelopmental factors and on neuronal projections emanating from the arcuate nucleus. The molecular and cellular anomalies detected in IUGR rats can be related to the reduced and delayed plasma leptin surge from d 0-16 when compared with control and IUGR rats with catch-up growth. However, the ability of leptin to activate intracellular signaling in arcuate nucleus neurons was not reduced in IUGR rats. Other mechanism such as epigenetic regulation of the major appetite-regulating neuropeptides genes was analyzed in parallel with their mRNA expression during postnatal development. This study reveals the importance of an early catch-up growth that reduces abnormal organization of hypothalamic pathways involved in energy homeostasis, whereas protein restriction, maintained during postnatal development leads to an important immaturity of the hypothalamus. PMID- 20016031 TI - Up-regulating the heme oxygenase system with hemin improves insulin sensitivity and glucose metabolism in adult spontaneously hypertensive rats. AB - Accumulating clinical evidence indicates that impaired glucose tolerance is a common phenomenon in essential hypertension. Although recent evidence underscores the role of heme-oxygenase (HO) in diabetes, its effects on insulin sensitivity and glucose metabolism in spontaneously hypertensive rat (SHR), a model of essential hypertension with characteristics of metabolic syndrome including insulin resistance/impaired glucose metabolism remains largely unclear. Here we report the effects of the HO inducer, hemin, and the HO blocker, chromium mesoporphyrin on insulin sensitivity and glucose metabolism in SHRs. Adult SHRs were severely hypertensive but normoglycemic. Hemin therapy lowered blood pressure, increased plasma insulin, decreased glycemia, and enhanced insulin sensitivity by improving glucose tolerance (ip glucose tolerance test) and insulin tolerance (ip insulin tolerance test) but reduced insulin resistance (homeostasis model assessment index). These effects were accompanied by increased gastrocnemius muscle HO-1, HO activity, cGMP, cAMP alongside antioxidants including bilirubin, ferritin, superoxide dismutase, catalase, and the total antioxidant capacity, whereas oxidative/inflammatory mediators like 8 isoprostane, nuclear-factor-kappaB, activating-protein-1, activating-protein-2, c Jun-NH2-terminal-kinase, and heme were abated. Furthermore, hemin reduced proteinuria/albuminuria and enhanced the depressed levels of adiponectin, AMP activated protein-kinase, and glucose transporter-4 in SHRs, suggesting that although SHRs are normoglycemic, insulin signaling and renal function may be impaired. Contrarily, the HO inhibitor chromium-mesoporphyrin exacerbated oxidative stress, aggravated insulin resistance, glucose tolerance, insulin tolerance and nephropathy. Hemin also enhanced HO signaling in Wistar Kyoto and Sprague Dawley rats and increased insulin sensitivity albeit less intensely than in SHRs, suggesting greater selectivity of HO in SHRs with dysfunctional insulin signaling. These results suggest that perturbations of insulin signaling may be a forerunner to hyperglycemia in essential hypertension. By concomitantly potentiating insulin-sensitizing agents, suppressing insulin/glucose intolerance, and abating oxidative stress, HO inducers may prevent metabolic and cardiovascular complications in essential hypertension. PMID- 20016032 TI - The unintended consequences of staffing mandates in Florida nursing homes: impacts on indirect-care staff. AB - Research on nursing staff ratios and quality of care in nursing homes prompted Florida to implement minimum nursing staff ratios for certified nursing assistants (CNAs) in 2001. Using the contingency theory, the authors investigated the response to this mandate and its potential effects on indirect-care staff. This study used the Online Survey, Certification, and Reporting (OSCAR) staffing data for freestanding Florida nursing homes between the years 1999 and 2004. Piecewise regression growth curve models were investigated to test whether the percentage of Medicaid residents is associated with change in indirect-care staffing levels. The number of indirect-care staff hours per 100 residents declined significantly following the mandated increase in nursing staff, particularly among facilities with a low percentage of Medicaid residents. This may have stemmed from a partial transfer of indirect-care to CNAs and was exacerbated in facilities that received less additional reimbursement to pay for CNA increases. PMID- 20016033 TI - Acute canine distemper encephalitis is associated with rapid neuronal loss and local immune activation. AB - For most virus infections of the central nervous system (CNS), immune-mediated damage, the route of inoculation and death of infected cells all contribute to the pathology observed. To investigate the role of these factors in early canine distemper neuropathogenesis, we infected ferrets either intranasally or intraperitoneally with the neurovirulent canine distemper virus strain Snyder Hill. Regardless of the route of inoculation, the virus primarily targeted the olfactory bulb, brainstem, hippocampus and cerebellum, whereas only occasional foci were detected in the cortex. The infection led to widespread neuronal loss, which correlated with the clinical signs observed. Increased numbers of activated microglia, reactive gliosis and different pro-inflammatory cytokines were detected in the infected areas, suggesting that the presence and ultimate death of infected cells at early times after infection trigger strong local immune activation, despite the observed systemic immunosuppression. PMID- 20016034 TI - RNA elements within the 5' untranslated region of the West Nile virus genome are critical for RNA synthesis and virus replication. AB - RNA elements within the flavivirus genome may play essential regulatory roles during virus replication. Here, recombinant West Nile virus (WNV) NS5 protein was used in combination with WNV subgenomic RNA templates to establish in vitro RNA dependent RNA polymerase and RNA-binding assays. These assays identified mutations in the stem-loop A (SLA) region of the 5' untranslated region (5'UTR) altering NS5 RNA synthesis and RNA-binding capability. These mutations were then introduced into the full-length WNV genome by reverse genetics. Further analysis of the mutant viruses showed that deletion of nt 46-60, which disrupted the stem and side loop of SLA, greatly compromised virus replication, whereas mutations that destroyed the top loop of SLA required for RNA synthesis in vitro did not significantly alter virus replication. These results suggest that SLA present in the 5'UTR of WNV is essential for RNA synthesis in vitro and for virus replication. PMID- 20016035 TI - Role of host-specific amino acids in the pathogenicity of avian H5N1 influenza viruses in mice. AB - Recent large-scale sequence analyses revealed 'signature' amino acids at specific positions in viral proteins that distinguish human influenza viruses from avian viruses. To determine the role of these host lineage-specific amino acids in the pathogenicity of H5N1 avian influenza viruses, we generated mutant viruses possessing signature amino acids in the PB2, PA and NP proteins of human influenza isolates ('human-like amino acids') in the genetic background of an avian H5N1 virus, and tested their pathogenicity in mice. We found that some of these mutants exhibited enhanced pathogenicity in mice, suggesting the involvement of these host lineage-specific amino acids in the pathogenicity of H5N1 avian influenza viruses in mammals. PMID- 20016037 TI - Intraspecies diversity of SARS-like coronaviruses in Rhinolophus sinicus and its implications for the origin of SARS coronaviruses in humans. AB - The Chinese rufous horseshoe bat (Rhinolophus sinicus) has been suggested to carry the direct ancestor of severe acute respiratory syndrome (SARS) coronavirus (SCoV), and the diversity of SARS-like CoVs (SLCoV) within this Rhinolophus species is therefore worth investigating. Here, we demonstrate the remarkable diversity of SLCoVs in R. sinicus and identify a strain with the same pattern of phylogenetic incongruence (i.e. an indication of recombination) as reported previously in another SLCoV strain. Moreover, this strain possesses a distinctive 579 nt deletion in the nsp3 region that was also found in a human SCoV from the late-phase epidemic. Phylogenetic analysis of the Orf1 region suggested that the human SCoVs are phylogenetically closer to SLCoVs in R. sinicus than to SLCoVs in other Rhinolophus species. These findings reveal a closer evolutionary linkage between SCoV in humans and SLCoVs in R. sinicus, defining the scope of surveillance to search for the direct ancestor of human SCoVs. PMID- 20016036 TI - Emergence of H5N1 avian influenza viruses with reduced sensitivity to neuraminidase inhibitors and novel reassortants in Lao People's Democratic Republic. AB - Pandemic influenza viruses can emerge through continuous evolution and the acquisition of specific mutations or through reassortment. This study assessed the pandemic potential of H5N1 viruses isolated from poultry outbreaks occurring from July 2006 to September 2008 in the Lao People's Democratic Republic (PDR). We analyzed 29 viruses isolated from chickens and ducks and two from fatal human cases in 2007. Prior to 2008, all H5N1 isolates in Lao PDR were from clade 2.3.4; however, clade 2.3.2 was introduced in September 2008. Of greatest concern was the circulation of three isolates that showed reduced sensitivity to the neuraminidase (NA) inhibitor oseltamivir in an enzyme inhibition assay, each with different NA mutations - V116A, I222L and K150N, and a previously unreported S246N mutation. In addition, six isolates had an S31N mutation in the M2 protein, which conferred resistance to amantadine not previously reported in clade 2.3.4 viruses. Two H5N1 reassortants were isolated whose polymerase genes, PB1 and PB2, were homologous to those of Eurasian viruses giving rise to a novel H5N1 genotype, genotype P. All H5N1 viruses retained avian-like receptor specificity, but four had altered affinities for alpha2,3-linked sialic acid. This study shows that, in a genetically similar population of H5N1 viruses in Lao PDR, mutants emerged with natural resistance to antivirals and altered affinities for alpha2,3 linked sialic acids, together with reassortants with polymerase genes homologous to Eurasian viruses. These changes may contribute to the emergence of a pandemic influenza strain and are critical in devising surveillance strategies. PMID- 20016038 TI - Phylogeny and genotyping of recent avian low-pathogenic H5 subtype influenza viruses from French ducks. AB - H5 low-pathogenic avian influenza virus (LPAIV) has the potential to become highly pathogenic and to cause serious problems in animal and public health. AIV surveillance and characterization in both wild and domestic species is therefore necessary. In order to acquire molecular information and to identify possible reassortments in French viruses, we analysed the entire genome of five H5N3, three H5N2 and two H5N1 LPAIV, isolated in France between 2002 and 2008 mostly from captive ducks (free-range commercial poultry or decoy ducks). Some of the genome sequences showed atypical characteristics, such as an insertion of 1 aa in the PB1 protein of one H5N3, a highly truncated PB1-F2 protein (11 aa in length instead of 90 aa) in one H5N2, and an insertion of 8 aa in the NS1 protein of H5N1. These two last molecular characteristics have not been described previously. Phylogenetic analysis demonstrated that all genes of French LPAIV, except the closely related matrix protein genes, clustered within the Eurasian avian influenzavirus lineage and fell into at least two phylogenetic subgroups. In addition, the French H5 LPAIV were segregated into eight genotypes, suggesting that many reassortment events have occurred in H5 LPAIV in Europe. However, it is not known whether the reassortment events have occurred in wild waterfowl and/or in captive birds in direct or indirect contact with wild birds. PMID- 20016039 TI - Bovine herpesvirus-1 US3 protein kinase: critical residues and involvement in the phosphorylation of VP22. AB - The US3 gene product of bovine herpesvirus-1 (BoHV-1) is a protein kinase that is expressed early during infection and capable of autophosphorylation. By examining differentially labelled US3 moieties by co-immunoprecipitation, we demonstrated that the protein kinase interacts with itself in vitro, which supports autophosphorylation by US3. Based on its homology to other serine/threonine protein kinases, we defined two highly conserved lysines in US3, at position 195 within the ATP-binding pocket and at position 282 within the catalytic loop; altering either residue resulted in kinase-dead mutants, demonstrating that these two residues are critical for the catalytic activity of BoHV-1 US3. During immunoprecipitation experiments, US3 interacted weakly with VP22, another tegument protein of BoHV-1. Furthermore, VP22 co-localized with US3 inside the nucleus in BoHV-1-infected cells. In vitro kinase assays demonstrated that VP22 is phosphorylated not only by US3, but also by the cellular casein kinase 2 (CK2) protein. The selective CK2 protein kinase inhibitor, 2-dimethylamino-4,5,6,7 tetrabromo-1H-benzimidazole (DMAT) and the less specific CK2 inhibitor Kenpaullone reduced VP22 phosphorylation, while CK1, protein kinase C or protein kinase A inhibitors did not affect phosphorylation. When US3 was included with VP22 in the kinase assay in the presence of DMAT, a low level of VP22 phosphorylation was observed. These data demonstrate that BoHV-1 VP22 interacts with both CK2 and US3, and that CK2 is the major kinase phosphorylating VP22, with US3 playing a minor role. PMID- 20016040 TI - Complete genome sequence and taxonomic position of anguillid herpesvirus 1. AB - Eel herpesvirus or anguillid herpesvirus 1 (AngHV1) frequently causes disease in freshwater eels. The complete genome sequence of AngHV1 and its taxonomic position within the family Alloherpesviridae were determined. Shotgun sequencing revealed a 249 kbp genome including an 11 kbp terminal direct repeat that contains 7 of the 136 predicted protein-coding open reading frames. Twelve of these genes are conserved among other members of the family Alloherpesviridae and another 28 genes have clear homologues in cyprinid herpesvirus 3. Phylogenetic analyses based on amino acid sequences of five conserved genes, including the ATPase subunit of the terminase, confirm the position of AngHV1 within the family Alloherpesviridae, where it is most closely related to the cyprinid herpesviruses. Our analyses support a recent proposal to subdivide the family Alloherpesviridae into two sister clades, one containing AngHV1 and the cyprinid herpesviruses and the other containing Ictalurid herpesvirus 1 and the ranid herpesviruses. PMID- 20016041 TI - Minireview: Won't get fooled again: the nonmetabolic roles of peroxisome proliferator-activated receptors (PPARs) in the heart. AB - The peroxisome proliferator-activated receptor (PPAR) transcription factors are nuclear receptors initially identified for their key role in regulating metabolic processes. Recent studies designed to identify the role of PPARalpha, -beta, and gamma in vivo uncovered extrametabolic roles that may be less well known in the heart. In this review, we describe what is known about these extrametabolic roles of PPARs, including regulation of cardiac inflammation, extracellular matrix remodeling, oxidative stress, and regulation of cardiac hypertrophy. Lastly, we discuss the emerging role of PPARs in cell cycle regulation and angiogenesis in noncardiac systems that may be applicable to heart biology. Although this review primarily discusses the extrametabolic role of PPARalpha, the most studied PPAR isoform in the heart, we highlight where possible what is known about the unique and overlapping roles of the PPAR isoforms in terms of metabolic function. PMID- 20016042 TI - Cellular immune responses of preterm infants after vaccination with whole-cell or acellular pertussis vaccines. AB - Based on studies reporting specific antibody titers, it is recommended to vaccinate preterm infants against Bordetella pertussis according to their chronological age. However, as specific T-cell responses also are involved in the protection against B. pertussis, we have determined whether highly preterm infants (<31 weeks) are able to mount these immune responses during vaccination. Forty-eight premature infants were vaccinated at 2, 3, and 4 months of their chronological age with an acellular (Pa; n = 24) or a whole-cell (Pw; n = 24) tetravalent diphtheria-tetanus-pertussis-polio vaccine, and blood samples were collected at 2, 3, and 6 months of age. Most of the Pa- and Pw-vaccinated infants developed at 3 or 6 months of age a gamma interferon (IFN-gamma) response to the B. pertussis antigens, accompanied by an interleukin-5 (IL-5) and IL-13 secretion for the Pa-vaccinated infants. No association was found between a very low infant birth weight, the occurrence of severe infections, and corticosteroid treatment or the administration of gammaglobulins with a low level of antigen-induced IFN gamma secretion. We conclude that like full-term infants, most preterm infants are able to mount a specific cellular immune response to the administration of the first doses of an acellular or a whole-cell pertussis vaccine. PMID- 20016043 TI - Francisella tularensis T-cell antigen identification using humanized HLA-DR4 transgenic mice. AB - There is no licensed vaccine against the intracellular pathogen Francisella tularensis. The use of conventional mouse strains to screen protective vaccine antigens may be problematic, given the differences in the major histocompatibility complex (MHC) binding properties between murine and human antigen-presenting cells. We used engineered humanized mice that lack endogenous MHC class II alleles but that express a human HLA allele (HLA-DR4 transgenic [tg] mice) to identify potential subunit vaccine candidates. Specifically, we applied a biochemical and immunological screening approach with bioinformatics to select putative F. tularensis subsp. novicida T-cell-reactive antigens using humanized HLA-DR4 tg mice. Cell wall- and membrane-associated proteins were extracted with Triton X-114 detergent and were separated by fractionation with a Rotofor apparatus and whole-gel elution. A series of proteins were identified from fractions that stimulated antigen-specific gamma interferon (IFN-gamma) production, and these were further downselected by the use of bioinformatics and HLA-DR4 binding algorithms. We further examined the validity of this combinatorial approach with one of the identified proteins, a 19-kDa Francisella tularensis outer membrane protein (designated Francisella outer membrane protein B [FopB]; FTN_0119). FopB was shown to be a T-cell antigen by a specific IFN gamma recall assay with purified CD4(+) T cells from F. tularensis subsp. novicida DeltaiglC-primed HLA-DR4 tg mice and cells of a human B-cell line expressing HLA-DR4 (DRB1*0401) functioning as antigen-presenting cells. Intranasal immunization of HLA-DR4 tg mice with the single antigen FopB conferred significant protection against lethal pulmonary challenge with an F. tularensis subsp. holarctica live vaccine strain. These results demonstrate the value of combining functional biochemical and immunological screening with humanized HLA DR4 tg mice to map HLA-DR4-restricted Francisella CD4(+) T-cell epitopes. PMID- 20016044 TI - Evaluation of a caprine arthritis-encephalitis virus/maedi-visna virus indirect enzyme-linked immunosorbent assay in the serological diagnosis of ovine progressive pneumonia virus in U.S. sheep. AB - A caprine arthritis-encephalitis virus (CAEV)/maedi-visna virus (MVV) indirect enzyme-linked immunosorbent assay (iELISA) was validated with samples from U.S. sheep and by the use of radioimmunoprecipitation as the standard for comparison. The sensitivity and the specificity were 86.0% (+ or - 5.8%) and 95.9% (+ or - 2.9%), respectively. The iELISA format and phylogenetic differences based on the MVV gag sequence contribute to the reduced sensitivity. PMID- 20016046 TI - Accuracy of AccessBio Immunoglobulin M and Total Antibody Rapid Immunochromatographic Assays for the Diagnosis of Acute Scrub Typhus Infection. AB - Using archived samples, we assessed the diagnostic capacity of a rapid immunochromatographic test (ICT) for the detection of Orientia tsutsugamushi IgM and total antibodies to aid with the diagnosis of acute scrub typhus infection in febrile patients in Laos. The sensitivity and the specificity of the ICT for the detection of IgM were 96.8% (121/125 samples; 95% confidence interval [CI], 92.1 to 99.1%) and 93.3% (98/105 samples; 95% CI, 86.7 to 97.3%), respectively. For the detection of total antibodies, the sensitivity was 97.6% (122/125 samples; 95% CI, 93.1 to 99.5%), but the specificity was much lower, at 71.4% (75/105 samples; 95% CI, 61.8 to 79.8%). PMID- 20016045 TI - Rational design and evaluation of a multiepitope chimeric fusion protein with the potential for leprosy diagnosis. AB - Despite the reduction in the number of leprosy cases registered worldwide as a result of the widespread use of multidrug therapy, the number of new cases detected each year remains stable in many countries. This indicates that Mycobacterium leprae, the causative agent of leprosy, is still being transmitted and that, without an earlier diagnosis, transmission will continue and infection will remain a health problem. The current means of diagnosis of leprosy is based on the appearance of clinical symptoms, which in many cases occur after significant and irreversible nerve damage has occurred. Our recent work identified several recombinant antigens that are specifically recognized by leprosy patients. The goal of the present study was to produce and validate the reactivity of a chimeric fusion protein that possesses the antibody binding properties of several of these proteins. The availability of such a chimeric fusion protein will simplify future test development and reduce production costs. We first identified the antibody binding regions within our top five antigen candidates by performing enzyme-linked immunosorbent assays with overlapping peptides representing the amino acid sequences of each protein. Having identified these regions, we generated a fusion construct of these components (protein advances diagnostic of leprosy [PADL]) and demonstrated that the PADL protein retains the antibody reactivity of the component antigens. PADL was able to complement a protein that we previously produced (the leprosy IDRI [Infectious Disease Research Institute] diagnostic 1 [LID-1] protein) to permit the improved diagnosis of multibacillary leprosy and that had a good ability to discriminate patients with multibacillary leprosy from control individuals. A serological diagnostic test consisting of these antigens could be applied within leprosy control programs to reduce transmission and to limit the appearance of leprosy associated disabilities and stigmatizing deformities by directing treatment. PMID- 20016047 TI - Adipokine profile and urinary albumin excretion in isolated growth hormone deficiency. AB - BACKGROUND: GH deficiency (GHD) is often associated with cardiovascular risk factors, including abdominal fat accumulation, hypercholesterolemia, and increased C-reactive protein. Despite the presence of these risk factors, adults with congenital lifetime isolated GHD (IGHD) due to an inactivating mutation in the GHRH receptor gene do not have premature atherosclerosis. OBJECTIVE: The aim was to study the serum levels of adiponectin and leptin (antiatherogenic and atherogenic adipokine, respectively), and the urinary albumin excretion (UAE) in these IGHD individuals. DESIGN AND PATIENTS: We conducted a cross-sectional study of 20 IGHD individuals (seven males; age, 50.8 +/- 14.6 yr) and 22 control subjects (eight males; age, 49.9 +/- 11.5 yr). MAIN OUTCOME MEASURES: Anthropometric factors, body composition, blood pressure, serum adiponectin, leptin, and UAE were measured. RESULTS: Adiponectin was higher [12.8 (7.1) vs. 9.7 (5) ng/ml; P = 0.041] in IGHD subjects, whereas no difference was observed in leptin [7.3 (6.3) vs. 9.3 (18.7 ng/ml] and UAE [8.6 (13.8) vs. 8.5 (11.1) microg/min]. CONCLUSIONS: Subjects with lifetime untreated IGHD have an adipokine profile with high adiponectin and normal leptin levels that may delay vascular damage and lesions of the renal endothelium. PMID- 20016048 TI - Are there any sensitive and specific sex steroid markers for polycystic ovary syndrome? AB - CONTEXT: Despite the high prevalence of hyperandrogenemia, the principal biochemical abnormality in women with polycystic ovary syndrome (PCOS), a definitive endocrine marker for PCOS has so far not been identified. OBJECTIVE: To identify a tentative diagnostic marker for PCOS, we compared serum levels of sex steroids, their precursors, and main metabolites in women with PCOS and controls. DESIGN AND METHODS: In this cross-sectional study of 74 women with PCOS and 31 controls, we used gas and liquid chromatography/mass spectrometry to analyze serum sex steroid precursors, estrogens, androgens, and glucuronidated androgen metabolites; performed immunoassays of SHBG, LH, and FSH; and calculated the LH/FSH ratio. RESULTS: Androgens and estrogens, sex steroid precursors, and glucuronidated androgen metabolites were higher in women with PCOS than in controls. In multivariate logistic regression analyses, estrone and free testosterone were independently associated with PCOS. The odds ratios per sd increase were 24.2 for estrone [95% confidence interval (CI), 4.0-144.7] and 12.8 for free testosterone (95% CI, 3.1-53.4). In receiver operating characteristic analyses, the area under curve was 0.93 for estrone (95% CI, 0.88-0.98) and 0.91 for free testosterone (95% CI, 0.86-0.97), indicating high sensitivity and specificity. CONCLUSION: Women with PCOS have elevated levels of sex steroid precursors, estrogens, androgens, and glucuronidated androgen metabolites as measured with a specific and sensitive mass spectrometry-based technique. The combination of elevated estrone (>50 pg/ml) and free testosterone (>3.3 pg/ml) appeared to discriminate with high sensitivity and specificity between women with and without PCOS. PMID- 20016049 TI - Increased circulating pro-inflammatory cytokines and Th17 lymphocytes in Hashimoto's thyroiditis. AB - CONTEXT: Th17 lymphocytes play an important role in different chronic inflammatory and autoimmune conditions. AIM: The aim of the study was to explore the status of Th17 cells in patients with autoimmune thyroid diseases (AITD). DESIGN: We assessed the serum levels and in vitro synthesis of IL-17 and IL-22 and of different cytokines (IL-6, IL-15, and IL-23) involved in the differentiation of Th17 cells in the peripheral blood and thyroid glands of 26 patients with AITD, eight with Graves' disease, and 18 with Hashimoto's thyroiditis (HT) as well as 10 healthy controls. RESULTS: We found enhanced levels of T cells synthesizing IL-17 and IL-22 in the peripheral blood from AITD patients, mainly in those with HT. In addition, a stronger expression of IL-17 and IL-22 and an enhanced number of IL-23R(+) cells was detected in thyroid glands from HT patients compared with Graves' disease or controls. Furthermore, increased concentrations of IL-6 and IL-15 were detected in sera from HT patients, whereas serum levels of IL-23 tended to be higher in these patients. Finally, an enhanced in vitro differentiation of T lymphocytes into Th17 cells induced by IL-23/IL-6 was observed in AITD patients. Accordingly, a strong induction of RORC2 gene was detected in lymphocytes from HT patients when stimulated with IL-23. CONCLUSION: Our results indicate that there is an increased differentiation of Th17 lymphocytes and an enhanced synthesis of Th17 cytokines in AITD, mainly in HT. These phenomena may have an important role in the pathogenesis of thyroid autoimmunity. PMID- 20016050 TI - Development of a disease-specific quality of life questionnaire in Addison's disease. AB - CONTEXT: Patients with Addison's disease reproducibly self-report impairment in specific dimensions of general well-being questionnaires, suggesting particular deficiencies in health-related quality-of-life (HRQoL). OBJECTIVE: We sought to develop an Addison's disease-specific questionnaire (AddiQoL) that could better quantify altered well-being and treatment effects. Design, Setting, Patients, Intervention, and Outcomes: We reviewed the literature to identify HRQoL issues in Addison's disease and interviewed patients and their partners in-depth to explore various symptom domains. A list of items was generated, and nine expert clinicians and five expert patients assessed the list for impact and clarity. A preliminary questionnaire was presented to 100 Addison's outpatients; the number of items was reduced after analysis of the distribution of the responses. The final questionnaire responses were assessed by Cronbach's alpha and Rasch analysis. RESULTS AND INTERPRETATION: Published studies of HRQoL in Addison's disease indicated reduced vitality and general health perception and limitations in physical and emotional functioning. In-depth interviews of 14 patients and seven partners emphasized the impact of the disease on the emotional domain. Seventy HRQoL items were generated; after the expert consultation process and pretesting in 100 patients, the number of items was reduced to 36. Eighty-six patients completed the final questionnaire; the responses showed high internal consistency with Cronbach's alpha 0.95 and Person Separation Index 0.94 (Rasch analysis). CONCLUSIONS: We envisage AddiQoL having utility in trials of hormone replacement and management of patients with Addison's disease, analogous to similar questionnaires in GH deficiency (AGHDA) and acromegaly (AcroQoL). PMID- 20016051 TI - Associations among metabolic syndrome, ischemia, inflammatory, oxidatives, and lipids biomarkers. AB - CONTEXT: Metabolic syndrome (MS) is described as a cluster of cardiometabolic risk factors. Studies suggest that ischemia-modified albumin (IMA) is a biomarker of cardiovascular diseases. IMA levels could be associated with cardiometabolic risks and represent a possible indication of microvascular dysfunction in MS patients. OBJECTIVE: To confirm this possible association, we evaluated the association between IMA levels and MS. DESIGN: We performed a case-control study (32 healthy individuals and 74 subjects with MS) to evaluate the association between MS, IMA, and other biomarkers [high-sensitivity C-reactive protein (hs CRP), oxidized low-density lipoprotein (OxLDL), oxidized low-density lipoprotein autoantibodies (anti-OxLDL), IL-6, lipid profile, and glucose]. RESULTS: The MS group showed higher levels of IMA (0.618 +/- 0.1355) as well as higher levels of hs-CRP, OxLDL, anti-OxLDL, and IL-6 than did control subjects (IMA = 0.338 +/- 0.0486) (P < 0.01). Multivariate analysis showed that IMA and MS association was independent of sex, age, diabetes mellitus 2, and hypercholesterolemia. CONCLUSION: We found an association between IMA and MS. Additional studies including prospective genetic variation approaches need to be performed to help elucidate this association between IMA and MS and its potential clinical role. PMID- 20016052 TI - Pharmacokinetics, metabolism, and excretion of anacetrapib, a novel inhibitor of the cholesteryl ester transfer protein, in rats and rhesus monkeys. AB - The pharmacokinetics and metabolism of anacetrapib (MK-0859), a novel cholesteryl ester transfer protein inhibitor, were examined in rats and rhesus monkeys. Anacetrapib exhibited a low clearance in both species and a moderate oral bioavailability of approximately 38% in rats and approximately 13% in monkeys. The area under the plasma concentration-time curve in both species increased in a less than dose-proportional manner over an oral dose range of 1 to 500 mg/kg. After oral administration of [(14)C]anacetrapib at 10 mg/kg, approximately 80 and 90% of the radioactive dose was recovered over 48 h postdose from rats and monkeys, respectively. The majority of the administered radioactive dose was excreted unchanged in feces in both species. Biliary excretion of radioactivity accounted for approximately 15% and urinary excretion for less than 2% of the dose. Thirteen metabolites, resulting from oxidative and secondary glucuronic acid conjugation, were identified in rat and monkey bile. The main metabolic pathways consisted of O-demethylation (M1) and hydroxylation on the biphenyl moiety (M2) and hydroxylation on the isopropyl side chain (M3); these hydroxylations were followed by O-glucuronidation of these metabolites. A glutathione adduct (M9), an olefin metabolite (M10), and a propionic acid metabolite (M11) also were identified. In addition to parent anacetrapib, M1, M2, and M3 metabolites were detected in rat but not in monkey plasma. Overall, it appears that anacetrapib exhibits a low-to-moderate degree of absorption after oral dosing and majority of the absorbed dose is eliminated via oxidation to a series of hydroxylated metabolites that undergo conjugation with glucuronic acid before excretion into bile. PMID- 20016053 TI - Metabolism and excretion of anacetrapib, a novel inhibitor of the cholesteryl ester transfer protein, in humans. AB - Anacetrapib is a novel cholesteryl ester transfer protein inhibitor being developed for the treatment of primary hypercholesterolemia and mixed dyslipidemia. The absorption, distribution, metabolism, and excretion of anacetrapib were investigated in an open-label study in which six healthy male subjects received a single oral dose of 150 mg and 165 microCi of [(14)C]anacetrapib. Plasma, urine, and fecal samples were collected at predetermined times for up to 14 days postdose and were analyzed for total radioactivity, the parent compound, and metabolites. The majority of the administered radioactivity (87%) was eliminated by fecal excretion, with negligible amounts present in urine (0.1%). The peak level of radioactivity in plasma (approximately 2 microM equivalents of [(14)C]anacetrapib) was achieved approximately 4 h postdose. The parent compound was the major radioactive component (79-94% of total radioactivity) in both plasma and feces. Three oxidative metabolites, M1, M2, and M3, were detected in plasma and feces and were identified as the O-demethylated species (M1) and two secondary hydroxylated derivatives of M1 (M2 and M3). Each metabolite was detected at low levels, representing or=1 year and 60 parent participants were recruited from 2 pediatric diabetes centers. Mentors were trained to provide social support (home visits and phone calls) for 12 months to families in the experimental arm (32 mothers). Control group parents (28 mothers) received the phone number of an experienced parent (not trained to give social support) to call as needed. Findings Mothers in the experimental and control arms differed at baseline only in birth order of the child with T1DM. The 2 groups did not differ significantly at 3, 6, or 12 months in parent concern, confidence, worry, impact on the family, or perceived social support. Mothers in the experimental arm identified the parent mentor as someone they would seek for advice and issues regarding growth and development, sleep, eating habits, and identification of community agencies. Parent mentors consistently referred mothers to health care providers for advice on medications and treatments but helped them incorporate this advice into day-to-day management. CONCLUSION: Mothers in the experimental arm valued the mentors' help in adjusting to the diagnosis, but this value was not measured by the study instruments. Focus group research is under way to clarify the concept of parent mentor social support and to develop a social support measurement tool. PMID- 20016059 TI - Wolves in sheep's clothing: SDO asymmetrically predicts perceived ethnic victimization among white and Latino students across three years. AB - Dominant groups have claimed to be the targets of discrimination on several historical occasions during violent intergroup conflict and genocide.The authors argue that perceptions of ethnic victimization among members of dominant groups express social dominance motives and thus may be recruited for the enforcement of group hierarchy. They examine the antecedents of perceived ethnic victimization among dominants, following 561 college students over 3 years from freshman year to graduation year. Using longitudinal, cross-lagged structural equation modeling, the authors show that social dominance orientation (SDO) positively predicts perceived ethnic victimization among Whites but not among Latinos, whereas victimization does not predict SDO over time. In contrast, ethnic identity and victimization reciprocally predicted each other longitudinally with equal strength among White and Latino students. SDO is not merely a reflection of contextualized social identity concerns but a psychological, relational motivation that undergirds intergroup attitudes across extended periods of time and interacts with the context of group dominance. PMID- 20016060 TI - JCS in 2010 - ringing in the changes. PMID- 20016061 TI - Visualizing cellular processes at the molecular level by cryo-electron tomography. AB - The cellular landscape rapidly changes throughout the biological processes that transpire within a cell. For example, the cytoskeleton is remodeled within fractions of a second. Therefore, reliable structural analysis of the cell requires approaches that allow for instantaneous arrest of functional states of a given process while offering the best possible preservation of the delicate cellular structure. Electron tomography of vitrified but otherwise unaltered cells (cryo-ET) has proven to be the method of choice for three-dimensional (3D) reconstruction of cellular architecture at a resolution of 4-6 nm. Through the use of cryo-ET, the 3D organization of macromolecular complexes and organelles can be studied in their native environment in the cell. In this Commentary, we focus on the application of cryo-ET to study eukaryotic cells - in particular, the cytoskeletal-driven processes that are involved in cell movements, filopodia protrusion and viral entry. Finally, we demonstrate the potential of cryo-ET to determine structures of macromolecular complexes in situ, such as the nuclear pore complex. PMID- 20016062 TI - The intersections between O-GlcNAcylation and phosphorylation: implications for multiple signaling pathways. AB - A paradigm-changing discovery in biology came about when it was found that nuclear and cytosolic proteins could be dynamically glycosylated with a single O linked beta-N-acetylglucosamine (O-GlcNAc) moiety. O-GlcNAcylation is akin to phosphorylation: it occurs on serine and/or threonine side chains of proteins, and cycles rapidly upon cellular activation. O-GlcNAc and phosphate show a complex interplay: they can either competitively occupy a single site or proximal sites, or noncompetitively occupy different sites on a substrate. Phosphorylation regulates O-GlcNAc-cycling enzymes and, conversely, O-GlcNAcylation controls phosphate-cycling enzymes. Such crosstalk is evident in all compartments of the cell, a finding that is congruent with the fundamental role of O-GlcNAc in regulating nutrient- and stress-induced signal transduction. O-GlcNAc transferase is recruited to the plasma membrane in response to insulin and is targeted to substrates by forming transient holoenzyme complexes that have different specificities. Cytosolic O-GlcNAcylation is important for the proper transduction of signaling cascades such as the NFkappaB pathway, whereas nuclear O-GlcNAc is crucial for regulating the activity of numerous transcription factors. This Commentary focuses on recent findings supporting an emerging concept that continuous crosstalk between phosphorylation and O-GlcNAcylation is essential for the control of vital cellular processes and for understanding the mechanisms that underlie certain neuropathologies. PMID- 20016063 TI - Modulation of Wnt signaling by the nuclear localization of cellular FLIP-L. AB - Cellular FLIP (cFLIP) inhibits the apoptosis signaling initiated by death receptor ligation. We previously reported that a long form of cFLIP (cFLIP-L) enhances Wnt signaling via inhibition of beta-catenin ubiquitylation. In this report, we present evidence that cFLIP-L translocates into the nucleus, which could have a role in modulation of Wnt signaling. cFLIP-L has a functional bipartite nuclear localization signal (NLS) at the C-terminus. Wild-type cFLIP-L (wt-FLIP-L) localizes in both the nucleus and cytoplasm, whereas NLS-mutated cFLIP-L localizes predominantly in the cytoplasm. cFLIP-L also has a nuclear export signal (NES) near the NLS, and leptomycin B, an inhibitor of CRM1 dependent nuclear export, increases the nuclear accumulation of cFLIP-L, suggesting that it shuttles between the nucleus and cytoplasm. Expression of mutant cFLIP-L proteins with a deletion or mutations in the NLS and NES confers resistance to Fas-mediated apoptosis, as does wt-FLIP-L, but they do not enhance Wnt signaling, which suggests an important role of the C-terminus of cFLIP-L in Wnt-signaling modulation. When wt-FLIP-L is expressed in the cytoplasm by conjugation with exogenous NES (NES-FLIP-L), Wnt signaling is not enhanced, whereas the NES-FLIP-L increases cytoplasmic beta-catenin as efficiently as wt FLIP-L. cFLIP-L physically interacts with the reporter plasmid for Wnt signaling, but not with the control plasmid. These results suggest a role for nuclear cFLIP L in the modulation of Wnt signaling. PMID- 20016064 TI - Selective and uncoupled role of substrate elasticity in the regulation of replication and transcription in epithelial cells. AB - Actin cytoskeleton forms a physical connection between the extracellular matrix, adhesion complexes and nuclear architecture. Because tissue stiffness plays key roles in adhesion and cytoskeletal organization, an important open question concerns the influence of substrate elasticity on replication and transcription. To answer this major question, polyelectrolyte multilayer films were used as substrate models with apparent elastic moduli ranging from 0 to 500 kPa. The sequential relationship between Rac1, vinculin adhesion assembly, and replication becomes efficient at above 200 kPa because activation of Rac1 leads to vinculin assembly, actin fiber formation and, subsequently, to initiation of replication. An optimal window of elasticity (200 kPa) is required for activation of focal adhesion kinase through auto-phosphorylation of tyrosine 397. Transcription, including nuclear recruitment of heterogeneous nuclear ribonucleoprotein A1 (hnRNP A1), occurred above 50 kPa. Actin fiber and focal adhesion signaling are not required for transcription. Above 50 kPa, transcription was correlated with alphav-integrin engagement together with histone H3 hyperacetylation and chromatin decondensation, allowing little cell spreading. By contrast, soft substrate (below 50 kPa) promoted morphological changes characteristic of apoptosis, including cell rounding, nucleus condensation, loss of focal adhesions and exposure of phosphatidylserine at the outer cell surface. On the basis of our data, we propose a selective and uncoupled contribution from the substrate elasticity to the regulation of replication and transcription activities for an epithelial cell model. PMID- 20016065 TI - Human RBMY regulates germline-specific splicing events by modulating the function of the serine/arginine-rich proteins 9G8 and Tra2-{beta}. AB - RBMY is a male germline RNA binding protein and potential alternative splicing regulator, but the lack of a convenient biological system has made its cellular functions elusive. We found that human RBMY fused to green fluorescent protein was strictly nuclear in transfected cells, but spatially enriched in areas around nuclear speckles with some components of the exon junction complex (EJC). Human RBMY (hRBMY) and the EJC components Magoh and Y14 also physically interacted but, unlike these two proteins, hRBMY protein did not shuttle to the cytoplasm. In addition, it relocalised into nucleolar caps after inhibition of RNA polymerase II transcription. Protein interactions were also detected between RBMY and splicing factors 9G8 and transformer-2 protein homolog beta (Tra2-beta), mediated by multiple regions of the RBMY protein that contain serine/arginine-rich dipeptides, but not by the single region lacking such dipeptides. These interactions modulated the splicing of several pre-mRNAs regulated by 9G8 and Tra2-beta. Importantly, ectopic expression of hRBMY stimulated the inclusion of a testis-enriched exon from the Acinus gene, whereas 9G8 and Tra2-beta repressed this exon. We propose that hRBMY associates with regions of the nucleus enriched in nascent RNA and participates in the regulation of specific splicing events in the germline by modulating the activity of constitutively expressed splicing factors. PMID- 20016067 TI - Sonic-hedgehog-mediated proliferation requires the localization of PKA to the cilium base. AB - Cerebellar granular neuronal precursors (CGNPs) proliferate in response to the mitogenic activity of Sonic hedgehog (Shh), and this proliferation is negatively regulated by activation of cAMP-dependent protein kinase (PKA). In the basal state, the PKA catalytic subunits (C-PKA) are inactive because of their association with the regulatory subunits (R-PKA). As the level of cAMP increases, it binds to R-PKA, displacing and thereby activating the C-PKA. Here we report that, in the presence of Shh, inactive C-PKA accumulates at the cilium base of proliferative CGNPs whereas removal of Shh triggers the activation of PKA at this particular location. Furthermore, we demonstrate that the anchoring of the PKA holoenzyme to the cilium base is mediated by the specific binding of the type II PKA regulatory subunit (RII-PKA) to the A-kinase anchoring proteins (AKAPs). Disruption of the interaction between RII-PKA and AKAPs inhibits Shh activity and, therefore, blocks proliferation of CGNP cultures. Collectively, these results demonstrate that the pool of PKA localized to the cilium base of CGNP plays an essential role in the integration of Shh signal transduction. PMID- 20016066 TI - Integrin activation by Fam38A uses a novel mechanism of R-Ras targeting to the endoplasmic reticulum. AB - The integrin family of heterodimeric cell-surface receptors are fundamental in cell-cell and cell-matrix adhesion. Changes to either integrin-ligand affinity or integrin gene expression are central to a variety of disease processes, including inflammation, cardiovascular disease and cancer. In screening for novel activators of integrin-ligand affinity we identified the previously uncharacterised multi-transmembrane domain protein Fam38A, located at the endoplasmic reticulum (ER). siRNA knockdown of Fam38A in epithelial cells inactivates endogenous beta1 integrin, reducing cell adhesion. Fam38A mediates integrin activation by recruiting the small GTPase R-Ras to the ER, which activates the calcium-activated protease calpain by increasing Ca(2+) release from cytoplasmic stores. Fam38A-induced integrin activation is blocked by inhibition of either R-Ras or calpain activity, or by siRNA knockdown of talin, a well-described calpain substrate. This highlights a novel mechanism for integrin activation by Fam38A, utilising calpain and R-Ras signalling from the ER. These data represent the first description of a novel spatial regulator of R-Ras, of an alternative integrin activation-suppression pathway based on direct relocalisation of R-Ras to the ER, and of a mechanism linking R-Ras and calpain signalling from the ER with modulation of integrin-ligand affinity. PMID- 20016068 TI - Long nuclear-retained non-coding RNAs and allele-specific higher-order chromatin organization at imprinted snoRNA gene arrays. AB - The imprinted Snurf-Snrpn domain, also referred to as the Prader-Willi syndrome region, contains two approximately 100-200 kb arrays of repeated small nucleolar (sno)RNAs processed from introns of long, paternally expressed non-protein-coding RNAs whose biogenesis and functions are poorly understood. We provide evidence that C/D snoRNAs do not derive from a single transcript as previously envisaged, but rather from (at least) two independent transcription units. We show that spliced snoRNA host-gene transcripts accumulate near their transcription sites as structurally constrained RNA species that are prevented from diffusing, as well as multiple stable nucleoplasmic RNA foci dispersed in the entire nucleus but not in the nucleolus. Chromatin structure at these repeated arrays displays an outstanding parent-of-origin-specific higher-order organization: the transcriptionally active allele is revealed as extended DNA FISH signals whereas the genetically identical, silent allele is visualized as singlet DNA FISH signals. A similar allele-specific chromatin organization is documented for snoRNA gene arrays at the imprinted Dlk1-Dio3 domain. Our findings have repercussions for understanding the spatial organization of gene expression and the intra-nuclear fate of non-coding RNAs in the context of nuclear architecture. PMID- 20016069 TI - Uncoupling of the spindle-checkpoint and chromosome-congression functions of BubR1. AB - The BubR1 checkpoint protein performs multiple functions in mitosis. We have carried out a functional analysis of conserved motifs of human BubR1 (also known as BUB1B) and demonstrate that spindle assembly checkpoint (SAC) and chromosome attachment functions can be uncoupled from each other. Mutation of five proline directed serine phosphorylation sites, identified in vivo by mass spectrometry, essentially abolishes attachment of chromosomes to the spindle but has no effect on SAC functionality. By contrast, mutation of the two conserved KEN boxes required for SAC function does not impact chromosome congression. Interestingly, the contribution of the two KEN-box motifs is not equal. Cdc20 associates with the N-terminal but not C-terminal KEN box, and mutation of the N-terminal KEN motif results in more severe acceleration of mitotic timing. Moreover, the two KEN motifs are not sufficient for maximal binding of Cdc20 and APC/C, which also requires sequences in the BubR1 C-terminus. Finally, mutation of the GLEBS motif causes loss of Bub3 interaction and mislocalization of BubR1 from the kinetochore; concomitantly, BubR1 phosphorylation as well as SAC activity and chromosome congression are impaired, indicating that the GLEBS motif is strictly required for both major functions of human BubR1. PMID- 20016070 TI - Distinct pools of cAMP centre on different isoforms of adenylyl cyclase in pituitary-derived GH3B6 cells. AB - Microdomains have been proposed to explain specificity in the myriad of possible cellular targets of cAMP. Local differences in cAMP levels can be generated by phosphodiesterases, which control the diffusion of cAMP. Here, we address the possibility that adenylyl cyclases, the source of cAMP, can be primary architects of such microdomains. Distinctly regulated adenylyl cyclases often contribute to total cAMP levels in endogenous cellular settings, making it virtually impossible to determine the contribution of a specific isoform. To investigate cAMP dynamics with high precision at the single-isoform level, we developed a targeted version of Epac2-camps, a cAMP sensor, in which the sensor was tagged to a catalytically inactive version of the Ca(2+)-stimulable adenylyl cyclase 8 (AC8). This sensor, and less stringently targeted versions of Epac2-camps, revealed opposite regulation of cAMP synthesis in response to Ca(2+) in GH(3)B(6) pituitary cells. Ca(2+) release triggered by thyrotropin-releasing hormone stimulated the minor endogenous AC8 species. cAMP levels were decreased by inhibition of AC5 and AC6, and simultaneous activation of phosphodiesterases, in different compartments of the same cell. These findings demonstrate the existence of distinct adenylyl cyclase-centered cAMP microdomains in live cells and open the door to their molecular micro-dissection. PMID- 20016071 TI - Direct demonstration of discrete Ca2+ microdomains associated with different isoforms of adenylyl cyclase. AB - Ca(2+)-sensitive adenylyl cyclases (ACs) orchestrate dynamic interplay between Ca(2+) and cAMP that is a crucial feature of cellular homeostasis. Significantly, these ACs are highly selective for capacitative Ca(2+) entry (CCE) over other modes of Ca(2+) increase. To directly address the possibility that these ACs reside in discrete Ca(2+) microdomains, we tethered a Ca(2+) sensor, GCaMP2, to the N-terminus of Ca(2+)-stimulated AC8. GCaMP2-AC8 measurements were compared with global, plasma membrane (PM)-targeted or Ca(2+)-insensitive AC2-targeted GCaMP2. In intact cells, GCaMP2-AC8 responded rapidly to CCE, but was largely unresponsive to other types of Ca(2+) rise. The global GCaMP2, PM-targeted GCaMP2 and GCaMP2-AC2 sensors reported large Ca(2+) fluxes during Ca(2+) mobilization and non-specific Ca(2+) entry, but were less responsive to CCE than GCaMP2-AC8. Our data reveal that different AC isoforms localize to distinct Ca(2+) microdomains within the plasma membrane. AC2, which is regulated via protein kinase C, resides in a microdomain that is exposed to a range of widespread Ca(2+) signals seen throughout the cytosol. By contrast, a unique Ca(2+) microdomain surrounds AC8 that promotes selectivity for Ca(2+) signals arising from CCE, and optimizes CCE-mediated cAMP synthesis. This direct demonstration of discrete compartmentalized Ca(2+) signals associated with specific signalling proteins provides a remarkable insight into the functional organization of signalling microdomains. PMID- 20016072 TI - Modulation of cell spreading and cell-substrate adhesion dynamics by dystroglycan. AB - Dystroglycan is a ubiquitously expressed cell adhesion protein. Its principal role has been determined as a component of the dystrophin-glycoprotein complex of muscle, where it constitutes a key component of the costameric cell adhesion system. To investigate more fundamental aspects of dystroglycan function in cell adhesion, we examined the role of dystroglycan in the dynamics and assembly of cellular adhesions in myoblasts. We show that beta-dystroglycan is recruited to adhesion structures and, based on staining for vinculin, that overexpression or depletion of dystroglycan affects both size and number of fibrillar adhesions. Knockdown of dystroglycan increases the size and number of adhesions, whereas overexpression decreases the number of adhesions. Dystroglycan knockdown or overexpression affects the ability of cells to adhere to different substrates, and has effects on cell migration that are consistent with effects on the formation of fibrillar adhesions. Using an SH3 domain proteomic screen, we identified vinexin as a binding partner for dystroglycan. Furthermore, we show that dystroglycan can interact indirectly with vinculin by binding to the vinculin-binding protein vinexin, and that this interaction has a role in dystroglycan-mediated cell adhesion and spreading. For the first time, we also demonstrate unequivocally that beta-dystroglycan is a resident of focal adhesions. PMID- 20016073 TI - N-WASP is a novel regulator of hair-follicle cycling that controls antiproliferative TGF{beta} pathways. AB - N-WASP is a cytoplasmic molecule mediating Arp2/3 nucleated actin polymerization. Mice with a keratinocyte-specific deletion of the gene encoding N-WASP showed normal interfollicular epidermis, but delayed hair-follicle morphogenesis and abnormal hair-follicle cycling, associated with cyclic alopecia and prolonged catagen and telogen phases. The delayed anagen onset correlated with an increased expression of the cell-cycle inhibitor p21CIP, and increased activity of the TGFbeta pathway, a known inducer of p21CIP expression. Primary N-WASP-null keratinocytes showed reduced growth compared with control cells and enhanced expression of the gene encoding the cell-cycle inhibitor p15INK4B, a TGFbeta target gene. Inhibition of TGFbeta signaling blocked overexpression of p15INK4B and restored proliferation of N-WASP-deficient keratinocytes in vitro. However, induction of N-WASP gene deletion in vitro did not result in obvious changes in TGFbeta signaling or growth of keratinocytes, indicating that the in vivo environment is required for the phenotype development. These data identify the actin nucleation regulator N-WASP as a novel element of hair-cycle control that modulates the antiproliferative and pro-apoptotic TGFbeta pathway in keratinocytes in vivo and in vitro. PMID- 20016074 TI - Integral membrane proteins Brr6 and Apq12 link assembly of the nuclear pore complex to lipid homeostasis in the endoplasmic reticulum. AB - Cells of Saccharomyces cerevisiae lacking Apq12, a nuclear envelope (NE) endoplasmic reticulum (ER) integral membrane protein, are defective in assembly of nuclear pore complexes (NPCs), possibly because of defects in regulating membrane fluidity. We identified BRR6, which encodes an essential integral membrane protein of the NE-ER, as a dosage suppressor of apq12 Delta. Cells carrying the temperature-sensitive brr6-1 allele have been shown to have defects in nucleoporin localization, mRNA metabolism and nuclear transport. Electron microscopy revealed that brr6-1 cells have gross NE abnormalities and proliferation of the ER. brr6-1 cells were hypersensitive to compounds that affect membrane biophysical properties and to inhibitors of lipid biosynthetic pathways, and displayed strong genetic interactions with genes encoding non essential lipid biosynthetic enzymes. Strikingly, brr6-1 cells accumulated, in or near the NE, elevated levels of the two classes of neutral lipids, steryl esters and triacylglycerols, and over-accumulated sterols when they were provided exogenously. Although neutral lipid synthesis is dispensable in wild-type cells, viability of brr6-1 cells was fully dependent on neutral lipid production. These data indicate that Brr6 has an essential function in regulating lipid homeostasis in the NE-ER, thereby impacting NPC formation and nucleocytoplasmic transport. PMID- 20016075 TI - Activity-dependent anchoring of importin alpha at the synapse involves regulated binding to the cytoplasmic tail of the NR1-1a subunit of the NMDA receptor. AB - Synaptic plasticity, the capacity of neurons to change the strength of their connections with experience, provides a mechanism for learning and memory in the brain. Long-term plasticity requires new transcription, indicating that synaptically generated signals must be transported to the nucleus. Previous studies have described a role for importin nuclear transport adaptors in mediating the retrograde transport of signals from synapse to nucleus during plasticity. Here, we investigated the possibility that stimulus-induced translocation of importins from synapse to nucleus involves activity-dependent anchoring of importins at the synapse. We show that importin alpha binds to a nuclear localization signal (NLS) present in the cytoplasmic tail of NR1-1a. This interaction is disrupted by activation of NMDA receptors in cultured neurons and by stimuli that trigger late-phase, but not early-phase, long-term potentiation of CA3-CA1 synapses in acute hippocampal slices. In vitro PKC phosphorylation of GST-NR1-1a abolishes its ability to bind importin alpha in brain lysates, and the interaction of importin alpha and NR1 in neurons is modulated by PKC activity. Together, our results indicate that importin alpha is tethered at the postsynaptic density by binding to the NLS present in NR1-1a. This interaction is activity dependent, with importin alpha being released following NMDA receptor activation and phosphorylation rendering it available to bind soluble cargoes and transport them to the nucleus during transcription-dependent forms of neuronal plasticity. PMID- 20016077 TI - ALCAM regulates mediolateral retinotopic mapping in the superior colliculus. AB - ALCAM [activated leukocyte cell adhesion molecule (BEN/SC-1/DM-GRASP)] is a transmembrane recognition molecule of the Ig superfamily (IgSF) containing five Ig domains (two V-type, three C2-type). Although broadly expressed in the nervous and immune systems, few of its developmental functions have been elucidated. Because ALCAM has been suggested to interact with the IgSF adhesion molecule L1, a determinant of retinocollicular mapping, we hypothesized that ALCAM might direct topographic targeting to the superior colliculus (SC) by serving as a substrate within the SC for L1 on incoming retinal ganglion cell (RGC) axons. ALCAM was expressed in the SC during RGC axon targeting and on RGC axons as they formed the optic nerve; however, it was downregulated distally on RGC axons as they entered the SC. Axon tracing with DiI revealed pronounced mistargeting of RGC axons from the temporal retina half of ALCAM null mice to abnormally lateral sites in the contralateral SC, in which these axons formed multiple ectopic termination zones. ALCAM null mutant axons were specifically compromised in medial orientation of interstitial branches, which is known to require the ankyrin binding function of L1. As a substrate, ALCAM-Fc protein promoted L1 dependent attachment of acutely dissociated retinal cells and an L1-expressing, ALCAM-negative cell line, consistent with an ALCAM-L1 heterophilic molecular interaction. Together, these results suggest a model in which ALCAM in the SC interacts with L1 on RGC axons to promote medial extension of RGC axon branches important for mediolateral axon targeting in the formation of retinocollicular maps. PMID- 20016076 TI - Selection and maintenance of spatial information by frontal eye field neurons. AB - Voluntary attention is often allocated according to internally maintained goals. Recent evidence indicates that the frontal eye field (FEF) participates in the deployment of spatial attention, even in the absence of saccadic eye movements. In addition, many FEF neurons maintain persistent representations of impending saccades. However, the role of persistent activity in the general maintenance of spatial information, and its relationship to spatial attention, has not been explored. We recorded the responses of single FEF neurons in monkeys trained to remember cued locations in order to detect changes in targets embedded among distracters in a task that did not involve saccades. We found that FEF neurons persistently encoded the cued location throughout the trial during the delay period, when no visual stimuli were present, and during visual discrimination. Furthermore, FEF activity reliably predicted whether monkeys would detect the target change. Population analyses revealed that FEF neurons with persistent activity were more effective at selecting the target from among distracters than neurons lacking persistent activity. These results demonstrate that FEF neurons maintain spatial information in the absence of saccade preparation and suggest that this maintenance contributes to the selection of relevant visual stimuli. PMID- 20016078 TI - Netrin-1-dependent spinal interneuron subtypes are required for the formation of left-right alternating locomotor circuitry. AB - Neuronal circuits in the spinal cord that produce the rhythmic and coordinated activities necessary for limb movements are referred to as locomotor central pattern generators (CPGs). The identities and preceding development of neurons essential for coordination between left and right limbs are not yet known. We show that the ventral floor plate chemoattractant Netrin-1 preferentially guides dorsally originating subtypes of commissural interneurons, the majority of which are inhibitory. In contrast, the excitatory and ventralmost V3 subtype of interneurons have a normal number of commissural fibers in Netrin-1 mutant mice, thus being entirely independent of Netrin-1-mediated attraction. This selective loss of commissural fibers in Netrin-1 mutant mice resulted in an abnormal circuitry manifested by a complete switch from alternating to synchronous fictive locomotor activity suggesting that the most ventral-originating excitatory commissural interneurons are an important component of a left-right synchrony circuit in the locomotor CPG. Thus, during development, Netrin-1 plays a critical role for the establishment of a functional balanced CPG. PMID- 20016079 TI - Neural and behavioral discrimination of sound duration by cats. AB - Sound duration is important for distinguishing auditory object. Previous studies on the neural representation of duration have usually lacked psychophysical data obtained from the same species; hence, the correspondence between neural and behavioral discrimination of duration remains obscure. We addressed this issue in cats by using the signal detection theory to investigate both neural activities in the primary auditory cortex (A1) and the cat's behavioral performance. We found that 320 ms duration can be well discriminated from 10 ms duration by some A1 neurons with specific response patterns: sustained response extended proportionally with the increase of stimulus duration and the On-Off response synchronizing stimulus onset and offset. Neurons with only On response cannot discriminate duration. The discrimination performance of both sustained and On Off responses deteriorated as the target duration decreased from 320 to 20 ms and the percentage of discriminative neurons (correct rate >0.75) decreased from 40 to 2%. Compared with the psychophysical results, we found that the psychometric functions of cats well matched the neurometric functions of most sustained response neurons and a small number of On-Off-response neurons. Pooling the spikes of multiple units improved neural discrimination, which may be attributable to the salience (noise reduction) of the responses in pooled data. Our results suggest that the sustained and Off responses of A1 neurons underlie the duration discrimination behavior of cats. PMID- 20016080 TI - Amyloid precursor protein regulates Cav1.2 L-type calcium channel levels and function to influence GABAergic short-term plasticity. AB - Amyloid precursor protein (APP) has been strongly implicated in the pathogenesis of Alzheimer's disease (AD). Although impaired synaptic function is believed to be an early and causative event in AD, how APP physiologically regulates synaptic properties remains poorly understood. Here, we report a critical role for APP in the regulation of L-type calcium channels (LTCC) in GABAergic inhibitory neurons in striatum and hippocampus. APP deletion in mice leads to an increase in the levels of Ca(v)1.2, the pore-forming subunit of LTCCs, and subsequent increases in GABAergic calcium currents (I(Ca(2+))) that can be reversed by reintroduction of APP. Upregulated levels of Ca(v)1.2 result in reduced GABAergic paired-pulse inhibition and increased GABAergic post-tetanic potentiation in both striatal and hippocampal neurons, indicating that APP modulates synaptic properties of GABAergic neurons by regulating Ca(v)1.2. Furthermore, APP physically interacts with Ca(v)1.2, suggesting a mechanism in which loss of APP leads to an inappropriate accumulation and aberrant activity of Ca(v)1.2. These results provide a direct link between APP and calcium signaling and might help explain how altered APP regulation leads to changes in synaptic function that occur with AD. PMID- 20016081 TI - Serotonin affects association of aversive outcomes to past actions. AB - Impairment in the serotonergic system has been linked to action choices that are less advantageous in a long run. Such impulsive choices can be caused by a deficit in linking a given reward or punishment with past actions. Here, we tested the effect of manipulation of the serotonergic system by tryptophan depletion and loading on learning the association of current rewards and punishments with past actions. We observed slower associative learning when actions were followed by a delayed punishment in the low serotonergic condition. Furthermore, a model-based analysis revealed a positive correlation between the length of the memory trace for aversive choices and subjects' blood tryptophan concentration. Our results suggest that the serotonergic system regulates the time scale of retrospective association of punishments to past actions. PMID- 20016082 TI - Frontal feedback-related potentials in nonhuman primates: modulation during learning and under haloperidol. AB - Feedback monitoring and adaptation of performance involve a medial reward system including medial frontal cortical areas, the medial striatum, and the dopaminergic system. A considerable amount of data has been obtained on frontal surface feedback-related potentials (FRPs) in humans and on the correlate of outcome monitoring with single unit activity in monkeys. However, work is needed to bridge knowledge obtained in the two species. The present work describes FRPs in monkeys, using chronic recordings, during a trial and error task. We show that frontal FRPs are differentially sensitive to successes and failures and can be observed over long-term periods. In addition, using the dopamine antagonist haloperidol we observe a selective effect on FRP amplitude that is absent for pure sensory-related potentials. These results describe frontal dopaminergic dependent FRPs in monkeys and corroborate a human-monkey homology for performance monitoring signals. PMID- 20016083 TI - Age- and gender-related differences in the cortical anatomical network. AB - Neuroanatomical differences attributable to aging and gender have been well documented, and these differences may be associated with differences in behaviors and cognitive performance. However, little is known about the dynamic organization of anatomical connectivity within the cerebral cortex, which may underlie population differences in brain function. In this study, we investigated age and sex effects on the anatomical connectivity patterns of 95 normal subjects ranging in age from 19 to 85 years. Using the connectivity probability derived from diffusion magnetic resonance imaging tractography, we characterized the cerebral cortex as a weighted network of connected regions. This approach captures the underlying organization of anatomical connectivity for each subject at a regional level. Advanced graph theoretical analysis revealed that the resulting cortical networks exhibited "small-world" character (i.e., efficient information transfer both at local and global scale). In particular, the precuneus and posterior cingulate gyrus were consistently observed as centrally connected regions, independent of age and sex. Additional analysis revealed a reduction in overall cortical connectivity with age. There were also changes in the underlying network organization that resulted in decreased local efficiency, and also a shift of regional efficiency from the parietal and occipital to frontal and temporal neocortex in older brains. In addition, women showed greater overall cortical connectivity and the underlying organization of their cortical networks was more efficient, both locally and globally. There were also distributed regional differences in efficiency between sexes. Our results provide new insights into the substrates that underlie behavioral and cognitive differences in aging and sex. PMID- 20016084 TI - Transplanted neural precursors enhance host brain-derived myelin regeneration. AB - In multiple sclerosis lesions resident oligodendrocyte progenitor cells (OPCs) are present, but fail to remyelinate. In the current study we examined whether neural precursor cell (NPC) transplantation can facilitate host brain-derived remyelination. We used the chronic cuprizone-induced demyelination model in aged mice, in which slow remyelination follows cuprizone removal. NPCs were transplanted to the lateral ventricles (intracerebroventricular) of cuprizone induced demyelinated brains. In this experimental setup, transplanted cells remained mostly in the periventricular area in an undifferentiated state. The extent of demyelination, remyelination, and proliferation of host brain regenerative cell population were examined at 1 week posttransplantation in the splenium of the corpus callosum, which was devoid of any transplanted cells. Transplantation of NPCs, but not of control, human embryonic kidney cells, significantly enhanced remyelination compared with sham-operated mice. Remyelination was performed exclusively by host brain OPCs. The proregenerative effect of transplanted NPCs was related to an increase in the proliferation of host brain OPCs. To examine the mechanism that underlies the proregenerative effect of NPCs in vitro, we used an NPC-OPC coculture system. These experiments indicated that NPCs induced the proliferation of OPCs and facilitated their differentiation into mature oligodendrocytes. The mitogenic effect of NPCs was mediated by platelet-derived growth factor-AA and fibroblast growth factor-2. In conclusion, NPC transplantation enhances host-derived myelin regeneration following chronic demyelination. This trophic effect may stimulate resident OPCs to overcome the remyelination failure in multiple sclerosis. PMID- 20016085 TI - Novel mediators of amyloid precursor protein signaling. AB - Multiple recent reports implicate amyloid precursor protein (APP) signaling in the pathogenesis of Alzheimer's disease, but the APP-dependent signaling network involved has not been defined. Here, we report a novel consensus sequence for interaction with the PDZ-1 and PDZ-2 domains of the APP-interacting proteins Mint1, Mint2, and Mint3 (X11alpha, X11beta, and X11gamma), and multiple novel interactors for these proteins, with the finding that transcriptional coactivators are highly represented among these interactors. Furthermore, we show that Mint3 interaction with a set of the transcriptional coactivators leads to nuclear localization and transactivation, whereas interaction of the same set with Mint1 or Mint2 prevents nuclear localization and transactivation. These results define new mediators of the signal transduction network mediated by APP. PMID- 20016086 TI - Deficiency of the 65 kDa isoform of glutamic acid decarboxylase impairs extinction of cued but not contextual fear memory. AB - Extinction procedures are clinically relevant for reducing pathological fear, and the mechanisms of fear regulation are a subject of intense research. The amygdala, hippocampus, and prefrontal cortex (PFC) have all been suggested to be key brain areas in extinction of conditioned fear. GABA has particularly been implicated in extinction learning, and the 65 kDa isoform of glutamic acid decarboxylase (GAD65) may be important in elevating GABA levels in response to environmental signals. Extinction of conditioned fear was examined in Gad65(-/-) mice while recording local field potentials from the amygdala, hippocampus, and PFC simultaneously while monitoring behavior. Gad65(-/-) mice showed generalization of cued fear, as reported previously, and impaired extinction of cued fear, such that fear remained high across extinction training. This endurance in cued fear was associated with theta frequency synchronization between the amygdala and hippocampus. Extinction of contextual fear, however, was unaltered in Gad65(-/-) mice when compared with wild-type littermates. The data imply that GAD65 plays a critical role in regulating cued fear responses during extinction learning and that, during this process, GABAergic signaling is involved in modulating synchronized activity between the amygdala and hippocampus. In view of the more pronounced effect on cued versus contextual fear extinction, these influences may rely more on GABAergic mechanisms in the amygdala. PMID- 20016087 TI - Orientation discrimination performance is predicted by GABA concentration and gamma oscillation frequency in human primary visual cortex. AB - Neuronal orientation selectivity has been shown in animal models to require corticocortical network cooperation and to be dependent on the presence of GABAergic inhibition. However, it is not known whether variability in these fundamental neurophysiological parameters leads to variability in behavioral performance. Here, using a combination of magnetic resonance spectroscopy, magnetoencephalography, and visual psychophysics, we show that individual performance on a visual orientation discrimination task is correlated with both the resting concentration of GABA and the frequency of stimulus-induced gamma oscillations in human visual cortex. Behaviorally, a strong oblique effect was found, with the mean angular threshold for oblique discrimination being five times higher than that for vertically oriented stimuli. Similarly, we found an oblique effect for the dependency of performance on neurophysiological parameters. Orientation detection thresholds were significantly negatively correlated with visual cortex GABA concentration for obliquely oriented patterns (r = -0.65, p < 0.015) but did not reach significance for vertically oriented stimuli (r = -0.39, p = 0.2). Similarly, thresholds for obliquely oriented stimuli were negatively correlated with gamma oscillation frequency (r = -0.65, p < 0.017), but thresholds for vertical orientations were not (r = -0.02, p = 0.9). Gamma oscillation frequency was positively correlated with GABA concentration in primary visual cortex (r = 0.67, p < 0.013). These results confirm the importance of GABAergic inhibition in orientation selectivity and demonstrate, for the first time, that interindividual performance on a simple visual task is linked to neurotransmitter concentration. The results also suggest a key role for GABAergic gamma oscillations in visual discrimination tasks. PMID- 20016088 TI - Overlapping and distinct neural systems code for subjective value during intertemporal and risky decision making. AB - During decision making, valuation of different types of rewards may involve partially distinct neural systems, but efficient choice behavior requires a common neural coding of stimulus value. We addressed this issue by measuring neural activity with functional magnetic resonance imaging while volunteers processed delayed and probabilistic decision options. Behaviorally, participants discounted both types of rewards in a hyperbolic manner, and discount rates, reflecting individual preferences, varied considerably between participants. Ventral striatum and orbitofrontal cortex showed a domain-general coding of subjective value regardless of whether rewards were delayed or probabilistic, strongly implicating these regions in the implementation of a common neural currency of value. In contrast, fronto-polar and lateral parietal cortex, as well as a region in the posterior cingulate cortex only correlated with the value of delayed rewards, whereas superior parietal cortex and middle occipital areas only represented the value of probabilistic rewards. These results suggest a mechanism for the neural coding of subjective value in the human brain that is based on the combination of domain-general and domain-specific valuation networks. PMID- 20016089 TI - Rescue of motor coordination by Purkinje cell-targeted restoration of Kv3.3 channels in Kcnc3-null mice requires Kcnc1. AB - The role of cerebellar Kv3.1 and Kv3.3 channels in motor coordination was examined with an emphasis on the deep cerebellar nuclei (DCN). Kv3 channel subunits encoded by Kcnc genes are distinguished by rapid activation and deactivation kinetics that support high-frequency, narrow action potential firing. Previously we reported that increased lateral deviation while ambulating and slips while traversing a narrow beam of ataxic Kcnc3-null mice were corrected by restoration of Kv3.3 channels specifically to Purkinje cells, whereas Kcnc3 mutant mice additionally lacking one Kcnc1 allele were partially rescued. Here, we report mice lacking all Kcnc1 and Kcnc3 alleles exhibit no such rescue. For Purkinje cell output to reach the rest of the brain it must be conveyed by neurons of the DCN or vestibular nuclei. As Kcnc1, but not Kcnc3, alleles are lost, mutant mice exhibit increasing gait ataxia accompanied by spike broadening and deceleration in DCN neurons, suggesting the facet of coordination rescued by Purkinje-cell-restricted Kv3.3 restoration in mice lacking just Kcnc3 is hypermetria, while gait ataxia emerges when additionally Kcnc1 alleles are lost. Thus, fast repolarization in Purkinje cells appears important for normal movement velocity, whereas DCN neurons are a prime candidate locus where fast repolarization is necessary for normal gait patterning. PMID- 20016091 TI - Diminutive digits discern delicate details: fingertip size and the sex difference in tactile spatial acuity. AB - We have observed that passive tactile spatial acuity, the ability to resolve the spatial structure of surfaces pressed upon the skin, differs subtly but consistently between the sexes, with women able to perceive finer surface detail than men. Eschewing complex central explanations, we hypothesized that this sex difference in somatosensory perception might result from simple physical differences between the fingers of women and men. To investigate, we tested 50 women and 50 men on a tactile grating orientation task and measured the surface area of the participants' index fingertips. In subsets of participants, we additionally measured finger skin compliance and optically imaged the fingerprint microstructure to count sweat pores. We show here that tactile perception improves with decreasing finger size, and that this correlation fully explains the better perception of women, who on average have smaller fingers than men. Indeed, when sex and finger size are both considered in statistical analyses, only finger size predicts tactile acuity. Thus, a man and a woman with fingers of equal size will, on average, enjoy equal tactile acuity. We further show that sweat pores, and presumably the Merkel receptors beneath them, are packed more densely in smaller fingers. PMID- 20016092 TI - Electrophysiology of object naming in primary progressive aphasia. AB - Primary progressive aphasia (PPA), a selective neurodegeneration of the language network, frequently causes object naming impairments. We examined the N400 event related potential (ERP) to explore interactions between object recognition and word processing in 20 PPA patients and 15 controls. Participants viewed photographs of objects, each followed by a word that was either a match to the object, a semantically related mismatch, or an unrelated mismatch. Patients judged whether word-object pairs matched with high accuracy (94% PPA group; 98% control group), but they failed to exhibit the normal N400 category effect (N400c), defined as a larger N400 to unrelated versus related mismatch words. In contrast, the N400 mismatch effect (N400m), defined as a larger N400 to mismatch than match words, was observed in both groups. N400m magnitude was positively correlated with neuropsychological measures of word comprehension but not fluency or grammatical competence, and therefore reflected the semantic component of naming. After ERP testing, patients were asked to name the same set of objects aloud. Trials with objects that could not be named were found to lack an N400m, although the name had been correctly recognized at the matching stage. Even accurate overt naming did not necessarily imply normal semantic processing, as shown by the absent N400c. The N400m was preserved in one patient with postsemantic anomia, who could write the names of objects she could not verbalize. N400 analyses can thus help dissect the multiple cognitive mechanisms that contribute to object naming failures in PPA. PMID- 20016090 TI - Early-life stress disrupts attachment learning: the role of amygdala corticosterone, locus ceruleus corticotropin releasing hormone, and olfactory bulb norepinephrine. AB - Infant rats require maternal odor learning to guide pups' proximity-seeking of the mother and nursing. Maternal odor learning occurs using a simple learning circuit including robust olfactory bulb norepinephrine (NE), release from the locus ceruleus (LC), and amygdala suppression by low corticosterone (CORT). Early life stress increases NE but also CORT, and we questioned whether early-life stress disrupted attachment learning and its neural correlates [2-deoxyglucose (2 DG) autoradiography]. Neonatal rats were normally reared or stressed-reared during the first 6 d of life by providing the mother with insufficient bedding for nest building and were odor-0.5 mA shock conditioned at 7 d old. Normally reared paired pups exhibited typical odor approach learning and associated olfactory bulb enhanced 2-DG uptake. However, stressed-reared pups showed odor avoidance learning and both olfactory bulb and amygdala 2-DG uptake enhancement. Furthermore, stressed-reared pups had elevated CORT levels, and systemic CORT antagonist injection reestablished the age-appropriate odor-preference learning, enhanced olfactory bulb, and attenuated amygdala 2-DG. We also assessed the neural mechanism for stressed-reared pups' abnormal behavior in a more controlled environment by injecting normally reared pups with CORT. This was sufficient to produce odor aversion, as well as dual amygdala and olfactory bulb enhanced 2-DG uptake. Moreover, we assessed a unique cascade of neural events for the aberrant effects of stress rearing: the amygdala-LC-olfactory bulb pathway. Intra-amygdala CORT or intra-LC corticotropin releasing hormone (CRH) infusion supported aversion learning with intra-LC CRH infusion associated with increased olfactory bulb NE (microdialysis). These results suggest that early-life stress disturbs attachment behavior via a unique cascade of events (amygdala-LC-olfactory bulb). PMID- 20016094 TI - Cooperative and competitive interactions facilitate stereo computations in macaque primary visual cortex. AB - Inferring depth from binocular disparities is a difficult problem for the visual system because local features in the left- and right-eye images must be matched correctly to solve this "stereo correspondence problem." Cortical architecture and computational studies suggest that lateral interactions among neurons could help resolve local uncertainty about disparity encoded in individual neurons by incorporating contextual constraints. We found that correlated activity among pairs of neurons in primary visual cortex depended both on disparity-tuning relationships and the stimuli displayed within the receptive fields of the neurons. Nearby pairs of neurons with distinct disparity tuning exhibited a decrease in spike correlation at competing disparities soon after response onset. Distant neuronal pairs of similar disparity tuning exhibited an increase in spike correlation at mutually preferred disparities. The observed correlated activity and response dynamics suggests that local competitive and distant cooperative interactions improve disparity tuning of individual neurons over time. Such interactions could represent a neural substrate for the principal constraints underlying cooperative stereo algorithms. PMID- 20016093 TI - Synchronous and asynchronous transmitter release at nicotinic synapses are differentially regulated by postsynaptic PSD-95 proteins. AB - The rate and timing of information transfer at neuronal synapses are critical for determining synaptic efficacy and higher network function. Both synchronous and asynchronous neurotransmitter release shape the pattern of synaptic influences on a neuron. The PSD-95 family of postsynaptic scaffolding proteins, in addition to organizing postsynaptic components at glutamate synapses, acts transcellularly to regulate synchronous glutamate release. Here we show that PSD-95 family members at nicotinic synapses on chick ciliary ganglion neurons in culture execute multiple functions to enhance transmission. Together, endogenous PSD-95 and SAP102 in the postsynaptic cell appear to regulate transcellularly the synchronous release of transmitter from presynaptic terminals onto the neuron while stabilizing postsynaptic nicotinic receptor clusters under the release sites. Endogenous SAP97, in contrast, has no effect on receptor clusters but acts transcellularly from the postsynaptic cell through N-cadherin to enhance asynchronous release. These separate and parallel regulatory pathways allow postsynaptic scaffold proteins to dictate the pattern of cholinergic input a neuron receives; they also require balancing of PSD-95 protein levels to avoid disruptive competition that can occur through common binding domains. PMID- 20016095 TI - Direct interaction of GABAB receptors with M2 muscarinic receptors enhances muscarinic signaling. AB - Downregulation of G-protein-coupled receptors (GPCRs) provides an important mechanism for reducing neurotransmitter signaling during sustained stimulation. Chronic stimulation of M(2) muscarinic receptors (M(2)Rs) causes internalization of M(2)R and G-protein-activated inwardly rectifying potassium (GIRK) channels in neuronal PC12 cells, resulting in loss of function. Here, we show that coexpression of GABA(B) R2 receptors (GBR2s) rescues both surface expression and function of M(2)R, including M(2)R-induced activation of GIRKs and inhibition of cAMP production. GBR2 showed significant association with M(2)R at the plasma membrane but not other GPCRs (M(1)R, mu-opioid receptor), as detected by fluorescence resonance energy transfer measured with total internal reflection fluorescence microscopy. Unique regions of the proximal C-terminal domains of GBR2 and M(2)R mediate specific binding between M(2)R and GBR2. In the brain, GBR2, but not GBR1, biochemically coprecipitates with M(2)R and overlaps with M(2)R expression in cortical neurons. This novel heteromeric association between M(2)R and GBR2 provides a possible mechanism for altering muscarinic signaling in the brain and represents a previously unrecognized role for GBR2. PMID- 20016096 TI - A novel allele of myosin VIIa reveals a critical function for the C-terminal FERM domain for melanosome transport in retinal pigment epithelial cells. AB - Mutations in the head and tail domains of the motor protein myosin VIIA (MYO7A) cause deaf-blindness (Usher syndrome type 1B, USH1B) and nonsyndromic deafness (DFNB2, DFNA11). The head domain binds to F-actin and serves as the MYO7A motor domain, but little is known about the function of the tail domain. In a genetic screen, we have identified polka mice, which carry a mutation (c.5742 + 5G > A) that affects splicing of the MYO7A transcript and truncates the MYO7A tail domain at the C-terminal FERM domain. In the inner ear, expression of the truncated MYO7A protein is severely reduced, leading to defects in hair cell development. In retinal pigment epithelial (RPE) cells, the truncated MYO7A protein is expressed at comparative levels to wild-type protein but fails to associate with and transport melanosomes. We conclude that the C-terminal FERM domain of MYO7A is critical for melanosome transport in RPE cells. Our findings also suggest that MYO7A mutations can lead to tissue-specific effects on protein levels, which may explain why some mutations in MYO7A lead to deafness without retinal impairment. PMID- 20016097 TI - Neuroprotective role of haptoglobin after intracerebral hemorrhage. AB - After intracerebral hemorrhage (ICH), the brain parenchyma is exposed to blood containing red blood cells (RBCs) and consequently to its lysis products. Iron rich hemoglobin (Hb) is the most abundant protein in RBCs. When released into the brain parenchyma during hemolysis, Hb becomes a central mediator of cytotoxicity. Our study indicates that haptoglobin (Hp), an acute-phase response protein primarily synthesized in the liver and known to bind and neutralize Hb in the bloodstream, is also expressed in brain in which it plays an important role in defending neurons from damage induced by hemolytic products after ICH. We demonstrate that the Hb-induced hypohaptoglobinemia aggravates ICH-induced brain damage while pharmacologic intervention with sulforaphane to induce brain Hp is linked to a reduction in brain damage. In agreement with these findings, Hp deficiency worsens whereas Hp overexpression alleviates ICH-mediated brain injury. We also identified that oligodendroglia are the primary source of brain derived Hp among brain cells and that oligodendroglia-released Hp plays protective roles against Hb-mediated toxicity to neurons and oligodendrocytes. We conclude that Hp, particularly the brain-derived Hp, plays cytoprotective roles and represents a potential therapeutic target for ICH treatment. PMID- 20016098 TI - Sustained morphine-induced sensitization and loss of diffuse noxious inhibitory controls in dura-sensitive medullary dorsal horn neurons. AB - Overuse of medications used to treat migraine headache can produce a chronic daily headache, termed medication overuse headache (MOH). Although "overuse" of opioids, triptans, and over-the-counter analgesics can all produce MOH, the neuronal mechanisms remain unknown. Headache pain is likely to be produced by stimulation of primary afferent neurons that innervate the intracranial vasculature and the resulting activation of medullary dorsal horn (MDH) neurons. The present study compared the receptive field properties of MDH dura-sensitive neurons in rats treated with morphine to those given vehicle. Animals were implanted with osmotic minipumps or pellets for sustained subcutaneous administration of morphine or vehicle 6-7 d before recording from dura-sensitive neurons. Electrical and mechanical activation thresholds from the dura were significantly lower in chronic morphine-treated animals when compared to vehicle controls. In addition, sustained morphine increased the cutaneous receptive field sizes. The presence of diffuse noxious inhibitory controls (DNICs) was examined by placing the tail in 55 degrees C water during concomitant noxious thermal stimulation of the cutaneous receptive field, usually located in the ophthalmic region. The DNIC stimulus produced significant inhibition of heat-evoked activity in vehicle- but not chronic morphine-treated animals. Inactivation of the rostral ventromedial medulla with 4% lidocaine reinstated DNICs in chronic morphine treated animals. These results are consistent with studies demonstrating a loss of DNICs in patients that suffer from chronic daily headache and may partially explain why overuse of medication used to treat migraine can induce headaches. PMID- 20016099 TI - GABA transporter-1 activity modulates hippocampal theta oscillation and theta burst stimulation-induced long-term potentiation. AB - The network oscillation and synaptic plasticity are known to be regulated by GABAergic inhibition, but how they are affected by changes in the GABA transporter activity remains unclear. Here we show that in the CA1 region of mouse hippocampus, pharmacological blockade or genetic deletion of GABA transporter-1 (GAT1) specifically impaired long-term potentiation (LTP) induced by theta burst stimulation, but had no effect on LTP induced by high-frequency stimulation or long-term depression induced by low-frequency stimulation. The extent of LTP impairment depended on the precise burst frequency, with significant impairment at 3-7 Hz that correlated with the time course of elevated GABAergic inhibition caused by GAT1 disruption. Furthermore, in vivo electrophysiological recordings showed that GAT1 gene deletion reduced the frequency of hippocampal theta oscillation. Moreover, behavioral studies showed that GAT1 knock-out mice also exhibited impaired hippocampus-dependent learning and memory. Together, these results have highlighted the important link between GABAergic inhibition and hippocampal theta oscillation, both of which are critical for synaptic plasticity and learning behaviors. PMID- 20016100 TI - Unexpected lack of hypersensitivity in LRRK2 knock-out mice to MPTP (1-methyl-4 phenyl-1,2,3,6-tetrahydropyridine). AB - Mutations in the leucine-rich repeat kinase 2 (LRRK2) gene are the most common known cause of Parkinson's disease (PD). Whether loss of LRRK2 function accounts for neurodegeneration of dopamine neurons in PD is not known, nor is it known whether LRRK2 kinase activity modulates the susceptibility of dopamine (DA) neurons to the selective dopaminergic toxin, 1-methyl-4-phenyl-1,2,3,6 tetrahydropyridine (MPTP). To better understand the role of LRRK2 in DA neuronal survival and its role in the susceptibility of DA neurons to MPTP, we generated LRRK2 knock-out (KO) mice lacking the kinase domain of LRRK2. Here, we show that LRRK2 KO mice are viable and have no major abnormalities and live to adulthood. The dopaminergic system is normal in LRRK2 KO mice as assessed via HPLC for DA and its metabolites and via stereologic assessment of DA neuron number in young and aged mice. Importantly, there is no significant difference in the susceptibility of LRRK2 KO and wild-type mice to MPTP. These results suggest that LRRK2 plays little if any role in the development and survival of DA neurons under physiologic conditions. Thus, PD due to LRRK2 mutations are likely not due to a loss of function. Moreover, LRRK2 is not required for the susceptibility of DA neurons to MPTP. PMID- 20016101 TI - Endolymphatic sodium homeostasis by extramacular epithelium of the saccule. AB - The saccule is a vestibular sensory organ that depends upon regulation of its luminal fluid, endolymph, for normal transduction of linear acceleration into afferent neural transmission. Previous studies suggested that endolymph in the saccule was merely derived from cochlear endolymph. We developed and used a preparation of isolated mouse saccule to measure transepithelial currents from the extramacular epithelium with a current density probe. The direction and pharmacology of transepithelial current was consistent with Na(+) absorption by the epithelial Na(+) channel (ENaC) and was blocked by the ENaC-specific inhibitors benzamil and amiloride. Involvement of Na(+),K(+)-ATPase and K(+) channels was demonstrated by reduction of the current by ouabain and the K(+) channel blockers Ba(2+), XE991, and 4-AP. Glucocorticoids upregulated the current via glucocorticoid receptors. Dexamethasone stimulated the current after 24 h and the stimulation was blocked by mifepristone but not spironolactone. No acute response was observed to elevated cAMP in the presence of amiloride nor to bumetanide, a blocker of Na(+),K(+),2Cl(-) cotransporter. The results are consistent with a canonical model of corticosteroid-regulated Na(+) absorption that includes entry of luminal Na(+) through apical membrane Na(+) channels and active basolateral exit of Na(+) via a Na(+) pump, with recycling of K(+) at the basolateral membrane via K(+)-permeable channels. These observations provide our first understanding of the active role played by saccular epithelium in the local regulation of the [Na(+)] of endolymph for maintenance of our sense of balance. PMID- 20016103 TI - Preparation to inhibit a response complements response inhibition during performance of a stop-signal task. AB - Inhibition of inappropriate responses is an essential executive function needed for adaptation to changing environments. In stop-signal tasks, which are often used to investigate response inhibition, subjects make "go" responses while they prepare to stop at a suddenly given "stop" signal. However, the preparatory processes ongoing before response inhibition have rarely been investigated, and it remains unclear how the preparation contributes to response inhibition. In the present study, a stop-signal task was designed so that the extent of the preparation could be estimated using behavioral and neuroimaging measures. Specifically, in addition to the conventional go trials where preparation to stop was required ("uncertain-go" trials), another type of go trial was introduced where a stop-signal was never given and such preparation was unnecessary ("certain-go" trials). An index reflecting the "preparation cost" was then calculated by subtracting the reaction times in the certain-go trials from those in the uncertain-go trials. It was revealed that the stop signal reaction time, a common index used to evaluate the efficiency of response inhibition, decreased as the preparation cost increased, indicating greater preparation supports more efficient inhibition. In addition, imaging data showed that response inhibition recruited frontoparietal regions (the contrast "stop vs uncertain-go") and that preparation recruited most of the inhibition-related frontoparietal regions (the contrast "uncertain-go vs certain-go"). It was also revealed that the inhibition related activity declined as the preparation cost increased. These behavioral and imaging results suggest preparation makes a complementary contribution to response inhibition during performance of a stop-signal task. PMID- 20016102 TI - The small GTPase Rac1 regulates auditory hair cell morphogenesis. AB - Morphogenesis of sensory hair cells, in particular their mechanotransduction organelle, the stereociliary bundle, requires highly organized remodeling of the actin cytoskeleton. The roles of Rho family small GTPases during this process remain unknown. Here we show that deletion of Rac1 in the otic epithelium resulted in severe defects in cochlear epithelial morphogenesis. The mutant cochlea was severely shortened with a reduced number of auditory hair cells and cellular organization of the auditory sensory epithelium was abnormal. Rac1 mutant hair cells also displayed defects in planar cell polarity and morphogenesis of the stereociliary bundle, including bundle fragmentation or deformation, and mispositioning or absence of the kinocilium. We further demonstrate that a Rac-PAK (p21-activated kinase) signaling pathway mediates kinocilium-stereocilia interactions and is required for cohesion of the stereociliary bundle. Together, these results reveal a critical function of Rac1 in morphogenesis of the auditory sensory epithelium and stereociliary bundle. PMID- 20016104 TI - Activity-dependent vesicular monoamine transporter-mediated depletion of the nucleus supports somatic release by serotonin neurons. AB - Packaging by the vesicular monoamine transporter (VMAT) is essential for mood controlling serotonin transmission but has not been assayed during activity. Here, two-photon imaging of the fluorescent serotonin analog 5,7 dihydroxytryptamine and three-photon imaging of endogenous serotonin were used to study vesicular packaging as it supports release from the soma of serotonin neurons. Glutamate receptor activation in dorsal raphe brain slice evoked somatic release that was mediated solely by vesicle exocytosis. This release was accompanied by VMAT-mediated serotonin depletion from the nucleus, a large compartment free of monoaminergic degradation pathways that has not been implicated in neurotransmission previously. Finally, while some monoamine packaged at rest was held in reserve, monoamine packaged during stimulation was released completely. Hence, somatic vesicles loaded by VMAT during activity rapidly undergo exocytosis. In the absence of active zones and with limited neurotransmitter reuptake, somatic release by serotonin neurons is supported by recruitment from a large pool of extravesicular serotonin in the nucleus and cytoplasm, and preferential release of the newly packaged transmitter. PMID- 20016105 TI - An intrinsic neuronal oscillator underlies dopaminergic neuron bursting. AB - Dopaminergic neurons of the ventral midbrain fire high-frequency bursts when animals are presented with unexpected rewards, or stimuli that predict reward. To identify the afferents that can initiate bursting and establish therapeutic strategies for diseases affected by altered bursting, a mechanistic understanding of bursting is essential. Our results show that bursting is initiated by a specific interaction between the voltage sensitivity of NMDA receptors and voltage-gated ion channels that results in the activation of an intrinsic, action potential-independent, high-frequency membrane potential oscillation. We further show that the NMDA receptor is uniquely suited for this because of the rapid kinetics and voltage dependence imparted to it by Mg(2+) ion block and unblock. This mechanism explains the discrete nature of bursting in dopaminergic cells and demonstrates how synaptic signals may be reshaped by local intrinsic properties of a neuron before influencing action potential generation. PMID- 20016106 TI - Auditory cortical activity after intracortical microstimulation and its role for sensory processing and learning. AB - Several studies have shown that animals can learn to make specific use of intracortical microstimulation (ICMS) of sensory cortex within behavioral tasks. Here, we investigate how the focal, artificial activation by ICMS leads to a meaningful, behaviorally interpretable signal. In natural learning, this involves large-scale activity patterns in widespread brain-networks. We therefore trained gerbils to discriminate closely neighboring ICMS sites within primary auditory cortex producing evoked responses largely overlapping in space. In parallel, during training, we recorded electrocorticograms (ECoGs) at high spatial resolution. Applying a multivariate classification procedure, we identified late spatial patterns that emerged with discrimination learning from the ongoing poststimulus ECoG. These patterns contained information about the preceding conditioned stimulus, and were associated with a subsequent correct behavioral response by the animal. Thereby, relevant pattern information was mainly carried by neuron populations outside the range of the lateral spatial spread of ICMS evoked cortical activation (approximately 1.2 mm). This demonstrates that the stimulated cortical area not only encoded information about the stimulation sites by its focal, stimulus-driven activation, but also provided meaningful signals in its ongoing activity related to the interpretation of ICMS learned by the animal. This involved the stimulated area as a whole, and apparently required large-scale integration in the brain. However, ICMS locally interfered with the ongoing cortical dynamics by suppressing pattern formation near the stimulation sites. The interaction between ICMS and ongoing cortical activity has several implications for the design of ICMS protocols and cortical neuroprostheses, since the meaningful interpretation of ICMS depends on this interaction. PMID- 20016107 TI - Adult plasticity in multisensory neurons: short-term experience-dependent changes in the superior colliculus. AB - Multisensory neurons in the superior colliculus (SC) have the capability to integrate signals that belong to the same event, despite being conveyed by different senses. They develop this capability during early life as experience is gained with the statistics of cross-modal events. These adaptations prepare the SC to deal with the cross-modal events that are likely to be encountered throughout life. Here, we found that neurons in the adult SC can also adapt to experience with sequentially ordered cross-modal (visual-auditory or auditory visual) cues, and that they do so over short periods of time (minutes), as if adapting to a particular stimulus configuration. This short-term plasticity was evident as a rapid increase in the magnitude and duration of responses to the first stimulus, and a shortening of the latency and increase in magnitude of the responses to the second stimulus when they are presented in sequence. The result was that the two responses appeared to merge. These changes were stable in the absence of experience with competing stimulus configurations, outlasted the exposure period, and could not be induced by equivalent experience with sequential within-modal (visual-visual or auditory-auditory) stimuli. A parsimonious interpretation is that the additional SC activity provided by the second stimulus became associated with, and increased the potency of, the afferents responding to the preceding stimulus. This interpretation is consistent with the principle of spike-timing-dependent plasticity, which may provide the basic mechanism for short term or long term plasticity and be operative in both the adult and neonatal SC. PMID- 20016108 TI - Nurr1 is required for maintenance of maturing and adult midbrain dopamine neurons. AB - Transcription factors involved in the specification and differentiation of neurons often continue to be expressed in the adult brain, but remarkably little is known about their late functions. Nurr1, one such transcription factor, is essential for early differentiation of midbrain dopamine (mDA) neurons but continues to be expressed into adulthood. In Parkinson's disease, Nurr1 expression is diminished and mutations in the Nurr1 gene have been identified in rare cases of disease; however, the significance of these observations remains unclear. Here, a mouse strain for conditional targeting of the Nurr1 gene was generated, and Nurr1 was ablated either at late stages of mDA neuron development by crossing with mice carrying Cre under control of the dopamine transporter locus or in the adult brain by transduction of adeno-associated virus Cre encoding vectors. Nurr1 deficiency in maturing mDA neurons resulted in rapid loss of striatal DA, loss of mDA neuron markers, and neuron degeneration. In contrast, a more slowly progressing loss of striatal DA and mDA neuron markers was observed after ablation in the adult brain. As in Parkinson's disease, neurons of the substantia nigra compacta were more vulnerable than cells in the ventral tegmental area when Nurr1 was ablated at late embryogenesis. The results show that developmental pathways play key roles for the maintenance of terminally differentiated neurons and suggest that disrupted function of Nurr1 and other developmental transcription factors may contribute to neurodegenerative disease. PMID- 20016110 TI - Choking vs. clutch performance: a study of sport performance under pressure. AB - Choking research in sport has suggested that an athlete's tendency to choke, versus give a better than usual (i.e., "clutch") performance depends on his or her personality, as well as on situational influences, such as a reliance on explicit (versus implicit) knowledge when pressured. The current study integrated these hypotheses and tested a structural equation model (SEM) to predict sport performance under pressure. Two hundred and one participants attempted two sets of 15 basketball free throws, and were videotaped during their second set of shots as a manipulation of pressure. Results of the model suggest that "reinvesting" attention in the task leads to greater anxiety (cognitive and somatic), which then predicts a higher level of self-focus; self-focus, then, did not lead to improved performance under pressure, whereas feelings of self reported "perceived control" did help performance. Implications for measurement of these constructs, and their relationships with performance, are discussed. PMID- 20016111 TI - Perfectionism, fear of failure, and affective responses to success and failure: the central role of fear of experiencing shame and embarrassment. AB - This study investigated how aspects of perfectionism in athletes (N = 388) related to the fears of failure proposed by Conroy et al. (2002), and how perfectionism and fears of failure predicted positive and negative affect after imagined success and failure in sports competitions. Results showed that perfectionistic personal standards showed a negative relationship with fear of experiencing shame and embarrassment and a positive relationship with positive affect after success, whereas perfectionistic concern over mistakes and perceived parental pressure showed a positive relationship with fear of experiencing shame and embarrassment and with negative affect after failure. Moreover, fear of experiencing shame and embarrassment fully mediated the relationship between perfectionistic concern and negative affect and between coach pressure and negative affect. The findings demonstrate that fear of experiencing shame and embarrassment is central in the relationship between perfectionism and fear of failure, and that perfectionistic concern about mistakes and perceived coach pressure are aspects of perfectionism that predict fear of experiencing shame and embarrassment and negative affect after failure. PMID- 20016109 TI - Emx1-lineage progenitors differentially contribute to neural diversity in the striatum and amygdala. AB - In the developing mammalian basal telencephalon, neural progenitors from the subpallium generate the majority of inhibitory medium spiny neurons (MSNs) in the striatum, while both pallial- and subpallial-derived progenitors contribute to excitatory and inhibitory neuronal diversity in the amygdala. Using a combination of approaches, including genetic fate mapping, cell birth dating, cell migration assays, and electrophysiology, we find that cells derived from the Emx1 lineage contribute to two distinct neuronal populations in the mature basal forebrain: inhibitory MSNs in the striatum and functionally distinct subclasses of excitatory neurons in the amygdala. Our cell birth-dating studies reveal that these two populations are born at different times during early neurogenesis, with the amygdala population born before the MSNs. In the striatum, Emx1-lineage neurons represent a unique subpopulation of MSNs: they are disproportionately localized to the dorsal striatum, are found in dopamine receiving, reelin positive patches, and are born throughout striatal neurogenesis. In addition, our data suggest that a subpopulation of these Emx1-lineage cells originate in the pallium and subsequently migrate to the developing striatum and amygdala. Our intersectional fate-mapping analysis further reveals that Emx1-lineage cells that coexpress Dlx exclusively generate MSNs but do not contribute to the excitatory neurons in the amygdala. Thus, both the timing of neurogenesis and differential combinatorial gene expression appear to be key determinants of striatal versus amygdala fate decisions of Emx1-lineage cells. PMID- 20016112 TI - Effect of acute exercise on cognitive control required during an Eriksen flanker task. AB - This study aimed to determine how cognitive control, engaged in a task requiring selective inhibition, is affected by acute steady-state exercise. An adapted version of the Eriksen flanker task, involving three types of trials that varied according to their level of congruency (congruent trials, stimulus-incongruent trials, and response-incongruent trials) was performed during 2 periods of 20-min cycling at a carefully controlled intensity (50% of maximal aerobic power). The results indicated that moderate exercise improves reaction time (RT) performance on the Eriksen flanker task. This facilitating effect appeared to be neither dependent on the nature of the interference (stimulus level conflict vs. response level conflict) nor on the amount of cognitive control engaged in the task (congruent vs. incongruent trials). Distributional RT analyses did not highlight any sign of impairment in the efficiency of cognitive control. PMID- 20016113 TI - Exploring the dose-response relationship between resistance exercise intensity and cognitive function. AB - The purpose of this study was to explore the dose-response relationship between resistance exercise intensity and cognitive performance. Sixty-eight participants were randomly assigned into control, 40%, 70%, or 100% of 10-repetition maximal resistance exercise groups. Participants were tested on Day 1 (baseline) and on Day 2 (measures were taken relative to performance of the treatment). Heart rate, ratings of perceived exertion, self-reported arousal, and affect were assessed on both days. Cognitive performance was assessed on Day 1 and before and following treatment on Day 2. Results from regression analyses indicated that there is a significant linear effect of exercise intensity on information processing speed, and a significant quadratic trend for exercise intensity on executive function. Thus, there is a dose-response relationship between the intensity of resistance exercise and cognitive performance such that high-intensity exercise benefits speed of processing, but moderate intensity exercise is most beneficial for executive function. PMID- 20016114 TI - Evidence of subliminally primed motivational orientations: the effects of unconscious motivational processes on the performance of a new motor task. AB - The aim of this study was to examine whether motivational orientations for a new motor task could be triggered by unconscious determinants. Participants were primed with subliminal words depicting an autonomous, a neutral, or a controlled motivation during an initial unrelated task, followed by working on an unknown motor task. Behavioral, physiological, and self-reported indicators of motivation for this task were assessed. Overall, results indicated a significant impact of the priming condition on all these indicators; whereas the priming of autonomous motivation led to positive outcomes, the priming of controlled motivation led to negative outcomes when compared with the neutral condition. Implications regarding the priming of unconscious determinants of motivation for sport and exercise are discussed. PMID- 20016115 TI - Knee bioelectric impedance assessment in healthy/with osteoarthritis subjects. AB - The present study analyzes parameters estimated by bioelectric impedance spectroscopy (BIS) in subjects with healthy and with osteoarthritis (OA) knees. Thirty-two male volunteers, members of the Parachute Military Infantry Brigade of Rio de Janeiro, Brazil, participated in the study (62 knee joints). Clinical specialists used the Dejour scale for OA classification and divided the subjects into a control (without OA) and a pathological group (with different degrees of OA). BIS data were obtained in a standing position using a BIS technique based on the current response to a step voltage excitation. Differences between groups were measured by means of a Wilcoxon-Mann-Whitney test. Results indicate that raw bioimpedance parameters seem to be sensitive to the physiological changes associated with OA. Thus, data indicate that extra-cellular resistance (Re) and reactance of the equivalent capacitance (Xcx) increase according to the disease intensity (p < 0.001). In conclusion, the BIS technique seems to be able to provide the objective and non-invasive basis for helping the diagnosis of knee OA. PMID- 20016116 TI - Full-field optical coherence tomography for the rapid estimation of epidermal thickness: study of patients with diabetes mellitus type 1. AB - Changes in morphology of the skin are an important factor that can affect non invasive measurements performed through this organ, in particular for glucose monitoring in e.g. patients with diabetes mellitus. A characterization technique for non-contact in vivo profiling of the superficial skin layers can be beneficial for evaluation of the performance of such measurement systems. We applied a full-field optical coherence tomography (OCT) system followed by the fully automatic processing for this task. With the developed procedure, non invasive quantification of the skin morphology can be performed within a few minutes. The dorsal skin of the upper arm of 22 patients with Type 1 Diabetes Mellitus was investigated with an OCT system and with a commercially available dermatological laser scanning confocal microscope (CM) as a reference method. The estimates of epidermal thickness from OCT were compared with the results of expert-assisted analysis of confocal images. The highest correlation with the CM measurements has been obtained for the distance from the entrance peak to the first minimum of the OCT reflection profile (R2 = 0.657, p < 0.0001). In this specific patient group, we have observed a statistically significant correlation of the subjects' body mass index with the distance from the entrance peak to the dermal reflection peak in the OCT profile (p = 0.010). Furthermore, the same OCT parameter is negatively correlated with age with marginal statistical significance (p = 0.062). At the same time, no relation of diabetes-related parameters (duration of disease and concentration of glycated haemoglobin) to the skin morphology observed with the OCT and CM was found. PMID- 20016117 TI - The effect of improved modelling of plasma clearance in paediatric patients with expanded body spaces on estimation of the glomerular filtration rate. AB - The glomerular filtration rate (GFR) is used clinically to assess renal function. The most accurate estimation technique is tracer clearance where deterministic compartment pharmacokinetic models are most widely used. The aim of this study was to assess the viability of alternative pharmacokinetic models to describe tracer clearance, and in turn, measure GFR. This study was carried out on 126 clearance datasets obtained from 44 patients with large solid tumours; these were fitted to four pharmacokinetic models with superiority of model determined by Akaike Information Criteria. A fractal model was found to be superior to the best deterministic compartment model (70% of datasets, P < 0.0020) as was a gamma distributed residence time model (93% of datasets, P < 0.0020); both models also gave greater mean weighted coefficients of determination than deterministic compartment models. These results suggest that gamma-distributed residence time and fractal models better describe tracer clearance than deterministic compartment models and therefore should allow more accurate estimation of GFR. PMID- 20016118 TI - Change of body movement coordination during cervical proprioceptive disturbances with increased age. AB - BACKGROUND: To date, there are very few studies on postural stability in older adults using body movement recordings to capture the postural movement pattern. Moreover, the importance of proprioception at key areas such as the calf or neck on the postural movement pattern in older adults has rarely been investigated. OBJECTIVE: To investigate whether the body movement coordination strategy to calf or neck vibration was affected by aging. METHODS: Body movement measurements were taken at five locations (ankle, knee, hip, shoulder and head) from 18 younger (mean age 29.1 years) and 16 older (mean age 71.5 years) adult subjects using a 3D movement measuring system while subjected to 50 s of pseudo-random calf or neck vibratory stimulation pulses with eyes open or closed. The positions from the knee, hip, shoulder and head markers were correlated against one another to give an indication of the body coordination. RESULTS: During quiet standing, older adults had greater correlation between the head and trunk than the young. There was an age effect in the body movement coordination strategy. Older adults had a different movement pattern with neck vibration involving mainly more independent knee movements, indicating balance difficulty. CONCLUSIONS: Neck vibration affects the movement pattern in older adults more compared with younger adults and calf vibration, suggesting that, the regulation of body orientation in older adults is more difficult, especially during cervical proprioceptive disturbances. PMID- 20016119 TI - A link between impaired lung function and increased cardiac stress. AB - BACKGROUND: Patients with impaired lung function often have systemic inflammation. C-reactive protein (CRP) and N-terminal pro-brain natriuretic peptide (NT-proBNP) are markers for inflammation and cardiac stress, respectively. OBJECTIVES: To evaluate the association between both markers and the potential impacts of lung disease on this relationship. METHODS: CRP and NT proBNP were prospectively measured in 697 consecutive outpatients (57.5 +/- 16.4 years) with chronic dyspnea. The patients were stratified into quartiles according to CRP levels (quartile 1: median CRP 0.35 mg/l; quartile 2: 1.50 mg/l; quartile 3: 3.62 mg/l; quartile 4: 10.90 mg/l) and classified into 2 categories based on the presence (n = 176) or absence (n = 521) of heart disease. RESULTS: Patients with at least moderately severe airway obstruction and those with interstitial lung disease had higher CRP values than patients without lung disease (median 3.50 vs. 4.34 vs. 1.80 mg/l, respectively; p < 0.01). In patients without heart disease, NT-proBNP values increased from CRP quartiles 1-3 to quartile 4 (median 47.4 vs. 82.1 pg/ml; p < 0.01) after adjusting for important covariates such as age, sex, body mass index, renal function and arterial hypertension. Likewise, the values for NT-proBNP were lower in CRP quartiles 1-3 than in quartile 4 (median 212.0 vs. 647.7 pg/ml; p < 0.01) in patients with heart disease after additional adjustment for the type of cardiac disorder. Lung disease had no direct effect on the relationship between CRP and NT-proBNP. CONCLUSION: Systemic inflammation that originates in the lung places an excess burden on the heart, which may contribute to the functional impairment of patients with advanced pulmonary disease. PMID- 20016120 TI - NIPAL4/ichthyin is expressed in the granular layer of human epidermis and mutated in two Pakistani families with autosomal recessive ichthyosis. AB - BACKGROUND: Autosomal recessive congenital ichthyosis (ARCI) can be divided into 3 types including lamellar ichthyosis (OMIM 242304), nonbullous congenital ichthyosiform erythroderma (OMIM 242100) and harlequin ichthyosis (OMIM 242500). The last type is uncommon since newborns with harlequin ichthyosis usually die shortly after birth. Several genes have been linked to ARCI, but these represent only 60% of the known genetic causes of this condition. METHODS: After having performed a linkage analysis, we analyzed the DNA of 2 consanguineous Pakistani families with ARCI for NIPAL4 mutations and performed in situ hybridization (ISH) for NIPAL4 mRNA in the epidermis. RESULTS: The haplotype analysis revealed a linkage to chromosome 5, and we identified a recurrent missense mutation, p.A176D, in affected individuals from both families. We also determined by ISH that NIPAL4 mRNA is highly expressed in the granular cell layer of the epidermis, consistent with the ARCI phenotype. CONCLUSION: Our results expand the spectrum of the clinical manifestations of the NIPAL4 gene and further extend our understanding of its molecular function. PMID- 20016121 TI - Temporal phase relation of circadian neural oscillations alters RFamide-related peptide-3 and testicular function in the mouse. AB - In order to study the effect of the temporal synergism of neural oscillations on reproductive regulation and the response of RFamide-related peptide-3 (RFRP-3; a mammalian ortholog of avian gonadotropin-inhibitory hormone), expression of immunoreactive RFRP-3 in the neurons of the dorsomedial nucleus of the hypothalamus was monitored in sexually immature and mature laboratory mice (study I). In study II, the effects of serotonin and dopamine precursors (5 hydroxytryptophan and L-dihydroxyphenylalanine; injected daily, 8 or 12 h apart, for 13 days in 3-week-old mice) on testicular activity and immunoreactive RFRP-3 neurons were studied until 24 days after treatment. Results indicate high levels of expression of immunoreactive RFRP-3 in the sexually immature and 8-hour mice (simulating gonadal suppression), while a low level was noted in mature and 12 hour mice (simulating gonadal stimulation). These findings not only suggest the modulation of gonadal development in mice (during the course of puberty attainment) by changing the temporal phase relation of serotonergic and dopaminergic oscillations (as in some seasonally breeding species), but also demonstrate an inverse correlation of RFRP-3 neurons and gonadal activity in both control and experimental conditions. PMID- 20016122 TI - Prevalence of percutaneous endoscopic gastrostomy in nursing home residents--a nationwide survey in Germany. AB - BACKGROUND: Despite extensive discussion about percutaneous endoscopic gastrostomy (PEG) in nursing home residents, no prevalence data exist in most countries. OBJECTIVE: This study aimed to determine the prevalence of PEG in German nursing homes, as well as nursing home characteristics that may influence this prevalence. Additionally, we documented the timing of PEG insertion, whether pre- or post-admission to the nursing home, and the proportion of residents receiving oral nutrition in addition to tube feeds. METHODS: A 26-item questionnaire was sent to 6,995 German nursing homes. Responses were anonymous. RESULTS: Of the 6,995 questionnaires sent out, 807 (11.5%) were completed and returned. Of the 66,507 total residents in the responding nursing homes, there were 4,390 residents with PEG (6.6%). The median PEG prevalence was 5.6% (5th 95th percentile 1.0-16.3%). Of all PEGs inserted, 55.3% were inserted before and 44.7% were inserted after nursing home admission. Oral beverage intake in addition to tube feeding was reported in 48.8% of residents with a PEG, and 43.5% of patients also received solid food. There were no significant differences in PEG prevalence between the federal states of Germany. The location (rural vs. urban) of the nursing home, its proprietor and food delivery method had no significant influence on PEG prevalence. With regard only to PEG tubes inserted after nursing home admission, all the factors stated above showed no significant influence on PEG prevalence. Weak positive correlations between PEG prevalence and the size of the nursing home as well as the number of total and certified nursing staff per resident could be detected, reflecting the heavy burden of care associated with residents requiring artificial nutrition. CONCLUSIONS: Compared to other countries, Germany can be assumed to have an average PEG prevalence in nursing home residents. No specific nursing home characteristics could be identified that affect PEG prevalence. PMID- 20016123 TI - Research on ageing, health and gender: A long and winding road. Reply to Mansdotter's 'Further thoughts on gender and lifetime health'. AB - This contribution is a reply to Dr. Mansdotter's comments on our discussion paper 'Health and well-being in old age: the pertinence of a gender mainstreaming approach in research' published in Gerontology [Gerontology 2010 (in press)]. Even though the comments are interesting and comprehensible, they cannot be left unanswered, this primarily because they are based on weak empirical evidence. (1) It is broadly uncontested that gender is not static. However, the conclusion that the more egalitarian division of parental duties can be viewed as an indicator for reduction of the gender gap in longevity and health is highly speculative. There is not enough empirical evidence to substantiate this position. (2) The 'caring hypothesis' proposed by Mansdotter, which holds that caring fathers develop less risky lifestyles and increased worries, is a possible, but not a sufficient explanation for gender convergence of physical and psychological health in future generations. Such a convergence seems to be heavily co determined by the changing lifestyles of women. (3) From a lifespan developmental perspective, androgyny does not mean gender equality, but a necessary openness of an individual for the positive traits of the opposing gender role, an essential trait for successful ageing. (4) Mansdotter's doubts concerning the implementation of gender mainstreaming in gerontological research and practice because of society's limited resources are not comprehensible. Exactly because economical resources are limited, and exactly because men and women have different resources and disadvantages due to their specific bio-psycho-social realities, the most efficient way of dealing with the gender gaps in health is with a differentialapproach. (5) The concluding recommendation of Mansdotter for more openness as a scientific position regarding the impact of gender roles on human health and well-being stands in contrast to her claim for normative standpoints and prioritization of either women or men in health promotion. We certainly need openness as a scientific position, but what we urgently need are good theoretical frameworks and more interdisciplinary and longitudinal approaches, which help to overcome the thin empirical base we have. This is a challenge and a chance for future research. PMID- 20016124 TI - Metacognitive training at home: does it improve older adults' learning? AB - BACKGROUND: Previous research has described the success of an intervention aimed at improving older adults' ability to regulate their learning. This metacognitive approach involves teaching older adults to allocate their study time more efficiently by testing themselves and restudying items that are less well learned. OBJECTIVE: Although this type of memory intervention has shown promise, training older adults to test themselves in the laboratory can be very time intensive. Thus, the purpose of the present study is to transport the self testing training method from the laboratory to home use. METHODS: A standard intervention design was used that included a pretraining session, multiple training sessions, and a posttraining session. Participants were randomly assigned to either the training group (n = 29) or the waiting list control group (n = 27). Moreover, we screened participants for whether they used the self testing strategy during their pretraining test session. RESULTS: Compared to the performance of the control group, the training group displayed significant gains, which demonstrates that older adults can benefit from training themselves to use these skills at home. Moreover, the results of the present study indicate that this metacognitive approach can effectively improve older adults' learning, even in those who spontaneously self-test prior to training. CONCLUSIONS: Training metacognitive skills, such as self-testing and efficient study allocation, can improve the ability to learn new information in healthy older adults. More importantly, older adult clients can be supplied with an at-home training manual, which will ease the burden on practitioners. PMID- 20016125 TI - Medical imaging of mummies and bog bodies--a mini-review. AB - Mummies are human remains with preservation of non-bony tissue. Mummification by natural influences results in so-called natural mummies, while mummification induced by active (human) intervention results in so-called artificial mummies, although many cultures practiced burial rites which to some degree involved both natural and artificial mummification. Since they are so uniquely well-preserved, mummies may give many insights into mortuary practices and burial rites. Specifically, the presence of soft tissues may expand the scope of paleopathological studies. Many recent mummy studies have focused on the development and application of non-destructive methods for examining mummies, especially radiography and CT scanning with advanced 3D visualizations. Indeed, the development of commercially available CT scanners in the 1970s meant that for the first time the 3D internal structure of mummies and bog bodies could be studied non-destructively. This article describes the history of mummy radiography and CT scanning, and some of the problems and opportunities involved in applying these techniques, derived for clinical use, on naturally and artificially preserved ancient human bodies. Unless severely degraded, bone is quite readily visualized, but accurate imaging of preserved soft tissues, and pathological lesions therein, may require considerable post-image capture processing of CT data. PMID- 20016126 TI - Nail matrix melanoma in situ: conservative surgical management. AB - BACKGROUND: Nail unit melanoma (NUM) is a rare variant of acral lentiginous melanoma. The differential diagnosis is wide but an acquired brown streak in the nail of a fair-skinned person must be considered a potential melanoma. Dermatoscopy helps clinicians to more accurately decide if a nail apparatus biopsy is necessary. METHODS: We report the case of a 61-year-old Caucasian woman with melanonychia occupying the central portion of the right thumbnail plate with 1 year of evolution. Dermatoscopy showed a brown pigmentation overlaid by longitudinal irregular lines. An excisional biopsy was performed, and pathological examination revealed melanoma in situ. For safety reasons, the nail unit was totally removed down to the phalangeal bone 3 weeks later, and a full thickness skin graft taken from the arm was used for reconstruction. CONCLUSION: NUMs pose a difficult treatment challenge. Wide excision with phalanx amputation is not satisfactory for patients with in situ and early invasive melanoma. Full thickness skin grafting after total nail unit excision is a simple procedure providing a good functional and cosmetic outcome. PMID- 20016127 TI - Ectoparasites from feral pigeons affecting humans. AB - Feral pigeons pose a considerable health risk to the human population. They are vectors of infectious diseases and source of antigens causing allergic diseases. Breeding and roosting sites of pigeons harbor parasites that may infest humans. In the present article, a concomitant parasitization of a young female with 3 different ectoparasites, the bedbug Cimex lectularius, the pigeon tick Argas reflexus and the red mite Dermanyssus gallinae, is reported. The parasites invaded the apartment from a balcony used as roost by feral pigeons and infested the patient continuously over a period of more than 2 months. To our knowledge this case presents the first record of a coincidental infestation of a single patient with several ectoparasite species deriving from feral pigeons. Additionally we report general symptoms in the patient probably caused by the high number of stings. Dermatologists should be aware of the possibility of an infestation with ectoparasites deriving from feral pigeons. In a review we give an overview on the most important ectoparasites transmitted from feral pigeons to humans and their importance for the dermatologist. PMID- 20016128 TI - Molecular characterization of the DDX3Y gene and its homologs in cattle. AB - DDX3Y (also known as DBY) is a member of the DEAD box protein family, which is involved in ATP-dependent RNA unwinding, needed in a variety of cellular processes including splicing, ribosome biogenesis and RNA degradation. In the human, DDX3Y is located in the AZFa interval in the Y chromosome. Deletion of the AZFa region has been shown to disrupt spermatogenesis, causing subfertility and infertility in otherwise healthy men. Here, we report the characterization of the bovine (b) DDX3Y gene and its homologs DDX3X and PL10. We found 2 transcripts for the bDDX3Y (bDDX3Y-L and -S), which correspond to the long and short transcripts of the human DDX3Y and mouse Ddx3y gene. The 2 transcripts are identical except for a 3-bp (AGT) insertion at the position of nt 2025 and an expanded 3'UTR (nt 2155-2769) in bDDX3Y-L. The bDDX3Y-S encodes a peptide of 660 amino acids (aa), while the bDDX3Y-L encodes a peptide of 661 aa as the result of an additional serine (S) insertion at the position of aa 634. Both bDDX3Y isoforms contain the conserved DEAD-box motif. The bDDX3Y is composed of 17 exons. The homologous gene on the X chromosome, bDDX3X, is highly conserved to the Y-copy at mRNA (83%) and protein (88%) levels as well as in the genomic structure. The autosomal copy, bPL10, mapped on BTA15, is a processed pseudogene with a similarity of 88.1% to bDDX3Y and 93.7% to bDDX3X mRNA, suggesting that PL10 is a retroposon of DDX3X. RT-PCR analyses showed that bDDX3Y-L, -S, bDDX3X and bPL10 were all widely expressed with predominant expression in testis and brain. Testicular section in situ hybridization revealed that sense and anti-sense RNAs of bDDX3Y-L, -S, and bDDX3X were expressed in interstitial cells. These results together with the finding that the pseudogene bPL10 is transcriptionally active in this study provide a basis for further investigating the DDX3 gene function in spermatogenesis, male fertility and gene evolution in mammals. PMID- 20016129 TI - Molecular cytogenetic characterization of a new wheat-Thinopyrum intermedium partial amphiploid resistant to powdery mildew and stripe rust. AB - A partial amphiploid, TE253, derived from crosses between the common wheat (Triticum aestivum L.) cultivar Yannong 15 and Thinopyrum intermedium (Host) Barkworth & D.R. Dewey was characterized by cytogenetic observations, disease resistance tests and genomic in situ hybridization (GISH). Mitotic observations showed that most plants of TE253 had 56 chromosomes, but a few had 54 or 55 chromosomes. The chromosomes in most pollen mother cells of plants with 2n = 56 formed 28 bivalents. Univalents (0.89 per cell) and tetravalents (0.087 per cell) occasionally occurred at meiotic metaphase I, showing a high degree of cytogenetic stability. After inoculation with the powdery mildew and stripe rust pathogens, Yannong 15 was highly susceptible, whereas TE253 and Th. intermedium were immune to both diseases. This indicated that the resistance of TE253 to powdery mildew and stripe rust was derived from Th. intermedium. GISH analysis using St-genomic DNA from Pseudoroegneria strigosa (M. Bieb) A. Love as a probe and ABD-genomic DNA from Chinese Spring wheat as a blocker demonstrated that TE253 consisted of 2 St-genome chromosomes, 8 J(S)-genome chromosomes, 2 SAT J chromosomes and 2 J-St translocated chromosomes. Line TE253 is a new partial amphiploid with resistance to both powdery mildew and stripe rust and can be used as a source of resistance genes in wheat improvement. PMID- 20016130 TI - A marsupial, Trichosurus vulpecula, DDX4/VASAGene (TvDDX4) of the DEAD box protein family: molecular conservation and germline expression. AB - The DDX4/VASA gene plays an important role in germ cell development in animals. We cloned and characterized a marsupial DDX4/VASA homolog (TvDDX4, 2,769 bps) from the possum and examined its expression in adult tissues at mRNA and protein levels. The isolated cDNA had a deduced 704 amino acid residues with significant homology to DDX4 from other animals, including mouse (86%) and human (87%). The DDX4 transcript was detected in the ovary and testis, and was undetectable in somatic tissues. The recombinant possum DDX4 protein (TvDDX4) was successfully produced in a bacterial expression system and used in polyclonal antibody generation. The recombinant TvDDX4 was detected by antibody against human DDX4 and mouse antibody against TvDDX4, but native possum TvDDX4 was only recognized by the possum antibody in the ovary and testis. Our results suggest a structural and functional conservation of DDX4 in marsupials and in mammals in the therian branches. PMID- 20016131 TI - Order out of chaos in the hybrid plant nucleus. AB - The plant nucleus is a highly ordered and dynamic structure, with a considerable level of variation between species in terms of genome size, genome organisation, chromosome territories and patterns associated with developmental changes. Diploids naturally represent the simplest state of affairs, but in the plant world more than 70% of species may have been involved in polyploidisation events at some stage during their evolution. Autopolyploids have multiple sets of chromosomes from a single species, and aside from the complexities of meiosis we may expect them to accommodate their polysomic state as well as their disomic relatives. Allopolyploids are at the other extreme, with multiple sets of chromosomes from 2 or more species, embedded in the cytoplasm of the maternal parent following hybridisation, and this presents the nucleus of nascent allopolyploids with certain zones of conflict. Nature has found ways to make the accommodation, and recent developments in molecular analysis have now opened a window for the experimenter to view the process of this adjustment, and to see how rapidly it takes place and what processes are involved. The nature of the resolution of nuclear conflicts in diploid hybrids and in allopolyploids is discussed. PMID- 20016132 TI - Sedation during flexible bronchoscopy in patients with pre-existing respiratory failure: Midazolam versus Midazolam plus Alfentanil. AB - BACKGROUND: The use of sedation during flexible bronchoscopy (FB) is undisputed; however, the combination of benzodiazepines and opiates, although reasonable, is suggested to cause hypoventilation, particularly in patients with pre-existing respiratory failure. OBJECTIVES: To assess respiratory function during FB. METHODS: Transcutaneous PCO(2 )(PtcCO(2)), oxygen saturation, patients' tolerance, time after FB until recovery and application of drug dosage were assessed in patients receiving either midazolam with alfentanil (n = 15) or midazolam alone (n = 15) for sedation for FB. RESULTS: There were no differences in PtcCO(2) values during FB between the two groups (all p > 0.05). However, PtcCO(2 )significantly increased over time in both groups (both p < 0.001; RM ANOVA on ranks). Minimum oxygen saturation (SaO(2)) [89 (interquartile range 79.8/92.8) vs. 86 (interquartile range 82.3/87.8)%; p = 0.46] and the duration until recovery, i.e., achieving an ALDRETE score of > or =9 [30 (interquartile range 10/90) vs. 10 (interquartile range 10/105) min; p = 0.68] were comparable for monosedation and combined sedation, respectively. The total amount of midazolam [4.0 (interquartile range 4.0/4.0) vs. 2.0 (interquartile range 2.0/2.0) mg; p < 0.001] was lower in patients receiving combined sedation. Significantly lower scores for pain and asphyxia, and a clear tendency to less nausea and cough were reported by patients receiving combined sedation. CONCLUSIONS: Combined sedation during FB produced a comparable degree of desaturation and hypoventilation, and is associated with a comparable time to full recovery compared to monosedation in patients with pre-existing respiratory failure. Importantly, FB using combined sedation is better tolerated by patients despite only 50% midazolam consumption. PMID- 20016133 TI - Copy number variation and missense mutations of the agouti signaling protein (ASIP) gene in goat breeds with different coat colors. AB - In goats, classical genetic studies reported a large number of alleles at the Agouti locus with effects on coat color and pattern distribution. From these early studies, the dominant A(Wt) (white/tan) allele was suggested to cause the white color of the Saanen breed. Here, we sequenced the coding region of the goat ASIP gene in 6 goat breeds (Girgentana, Maltese, Derivata di Siria, Murciano Granadina, Camosciata delle Alpi, and Saanen), with different coat colors and patterns. Five single nucleotide polymorphisms (SNPs) were identified, 3 of which caused missense mutations in conserved positions of the cysteine-rich carboxy terminal domain of the protein (p.Ala96Gly, p.Cys126Gly, and p.Val128Gly). Allele and genotype frequencies suggested that these mutations are not associated or not completely associated with coat color in the investigated goat breeds. Moreover, genotyping and sequencing results, deviation from Hardy-Weinberg equilibrium, as well as allele copy number evaluation from semiquantitative fluorescent multiplex PCR, indicated the presence of copy number variation (CNV) in all investigated breeds. To confirm the presence of CNV and evaluate its extension, we applied a bovine-goat cross-species array comparative genome hybridization (aCGH) experiment using a custom tiling array based on bovine chromosome 13. aCGH results obtained for 8 goat DNA samples confirmed the presence of CNV affecting a region of less that 100 kb including the ASIP and AHCY genes. In Girgentana and Saanen breeds, this CNV might cause the A(Wt) allele, as already suggested for a similar structural mutation in sheep affecting the ASIP and AHCY genes, providing evidence for a recurrent interspecies CNV. However, other mechanisms may also be involved in determining coat color in these 2 breeds. PMID- 20016134 TI - Alveolar cell senescence exacerbates pulmonary inflammation in patients with chronic obstructive pulmonary disease. AB - BACKGROUND: Alveolar cell senescence is accelerated in patients with chronic obstructive pulmonary disease (COPD). OBJECTIVES: We tested the hypothesis that alveolar cell senescence contributes to the chronic inflammation that affects the lungs of COPD patients. METHODS: We exposed alveolar type II-like epithelial (A549) cells to a G-quadruplex-interacting telomerase inhibitor in vitro to induce cellular senescence and analyzed the production of proinflammatory cytokines and the activation of NF-kappaB. Human dermal microvascular endothelial cells (HDMECs) were serially passaged to induce replicative senescence. We also immunostained human lung tissue sections obtained from COPD patients, asymptomatic smokers and asymptomatic nonsmokers and examined correlations between type II cell senescence and inflammation. RESULTS: Senescent A549 cells and HDMECs, whether stimulated with lipopolysaccharide or not, produced greater amounts of IL-6, IL-8 and TNF-alpha, which paralleled NF-kappaB activation, than did presenescent cells. There were positive correlations between the percentages of senescent type II cells that expressed p16(INK4a) and the percentages of type II cells that expressed phosphorylated NF-kappaB. The lung tissue of the COPD patients contained higher percentages of proinflammatory senescent type II cells that co-expressed p16(INK4a) and phosphorylated NF-kappaB than the tissue from asymptomatic smokers and asymptomatic nonsmokers. Higher percentages of p16(INK4a)-positive senescent type II cells than of p16(INK4a)-negative presenescent type II cells were positive for phosphorylated NF-kappaB. CONCLUSIONS: Senescence of alveolar epithelial cells is associated with functional alterations of the cells to a proinflammatory phenotype and may contribute to the pathogenesis of COPD. PMID- 20016135 TI - Growth hormone internalization in mitochondria decreases respiratory chain activity. AB - Growth hormone (GH) is a signaling molecule regulating cell proliferation, differentiation and metabolism via activation of specific cell surface receptors and subsequent triggering of signal transduction pathways. This is associated with GH/GH receptor internalization and accumulation of GH in several subcellular compartments, including mitochondria. To assess the functional relevance of such mitochondrial accumulation, we first confirmed the occurrence of mitochondrial GH uptake ex vivo as early as 10 min after (125)I-GH injection to the rats. We next showed that intact (125)I-GH accumulates in mitochondrial fractions in vitro in a specific, rapid and saturable manner with an apparent affinity (K(d)) of 1.44 nM. At the electron-microscopic level, immunoreactive GH density within mitochondria increased after in vitro hormone incubation, without any modification of the sub mitochondrial distribution pattern. The presence of GH in the inter-membrane space and at the inner membrane seen by electron microscopy was confirmed by SDS PAGE and autoradiography after mitochondrial fractioning thus suggesting the involvement of GH in the respiration control. To test this hypothesis further, we performed polarographic and spectrophotometric assays on isolated mitochondria. These assays pointed to a direct, selective and dose-dependent effect of GH on the inhibition of succinate dehydrogenase and cytochrome C oxidase activities. The latter inhibition was in contrast with indirect, GH receptor-initiated stimulation of cytochrome C oxidase activity observed in GH-treated whole BRL cells transfected to express this receptor. Altogether, these data show that GH is specifically imported in mitochondria, where it operates a direct metabolic effect, independently of cell surface receptors and signal transduction. PMID- 20016136 TI - Reversible ductus arteriosus constriction due to maternal indomethacin after fetal intervention for hypoplastic left heart syndrome with intact/restrictive atrial septum. AB - OBJECTIVE: Fetal cardiac intervention (FCI) has been performed at our center in selected fetuses with complex congenital heart disease since 2000. Most interventions are performed in fetuses with a ductus arteriosus (DA)-dependent circulation. Indomethacin promotes closure of the DA in newborns and in fetal life, a potentially life threatening complication in fetuses with ductus dependent congenital heart disease. METHODS: We reviewed our experience with FCI with a focus on the frequency, features, and clinical course of ductal constriction. Fetuses undergoing FCI receive comprehensive pre- and postoperative cardiac and cerebral ultrasound evaluation, approximately 24 hours before and after the procedure, including imaging of DA flow and Doppler assessment of the umbilical artery and vein, ductus venosus, and, since 2004, the middle cerebral artery. RESULTS: Among 113 fetuses that underwent FCI, 24 of which were older than 28 0/7 weeks gestation, 2 were found to have DA constriction due to indomethacin therapy within 24 hours of intervention. Both of these were 30-week fetuses with hypoplastic left heart syndrome and restrictive or intact atrial septum. The DA was stenotic by spectral and color Doppler, and middle cerebral and umbilical artery pulsatility indexes were depressed. After discontinuation of indomethacin, the Doppler indices improved or normalized. CONCLUSION: Close echocardiographic monitoring of fetal Doppler flow velocities is very important after fetal intervention and indomethacin treatment, as the consequences of DA constriction in a fetus with hypoplastic left heart syndrome are potentially lethal. Sonographic evaluation should include measurement of cerebral and umbilical arterial flow velocities as well as color and spectral Doppler interrogation of the DA. PMID- 20016138 TI - Are bronchioles fueling burning alveoli in lung fibrosis? PMID- 20016139 TI - Differentiation of Y chromosome in the X(1)X(1)X(2)X(2)/X(1)X(2)Y sex chromosome system of Hoplias malabaricus (Characiformes, Erythrinidae). AB - Six populations of Hoplias malabaricus from the Lower Paranapanema River were studied and showed 2n = 40 with metacentric/submetacentric chromosomes for females and 2n = 39 with m/sm chromosomes for males, thereby revealing a multiple sex chromosome system of the X(1)X(1)X(2)X(2)/X(1)X(2)Y type. The Y chromosome was the largest metacentric chromosome in all karyotypes. The heterochromatic blocks were located in a pericentromeric region of all pairs and in telomeric regions in some other pairs. A conspicuous pericentromeric C-band was detected in a single pair, equivalent to the X(1) chromosome. No markers on X(2) chromosomes were detected, but they could be identified by their size. The females displayed the same distribution of C and chromomycin A(3) bands and 18S rDNA sites among the populations. However, the males exhibited differences in both number and position of Giemsa C-bands on the Y chromosome, which were also evident after Chromomycin A(3) banding and FISH with the 18S rDNA probe. These cytogenetic tools allowed for the identification of possible mechanisms involved in the differentiation and evolution of the Y chromosome. PMID- 20016140 TI - Development of a reliable PCR-RFLP assay for investigation of the JAK2 rs10974944 SNP, which might predispose to the acquisition of somatic mutation JAK2(V617F). PMID- 20016137 TI - Telomeres and immunological diseases of aging. AB - A defining feature of the eukaryotic genome is the presence of linear chromosomes. This arrangement, however, poses several challenges with regard to chromosomal replication and maintenance. To prevent the loss of coding sequences and to suppress gross chromosomal rearrangements, linear chromosomes are capped by repetitive nucleoprotein structures, called telomeres. Each cell division results in a progressive shortening of telomeres that, below a certain threshold, promotes genome instability, senescence, and apoptosis. Telomeric erosion, maintenance, and repair take center stage in determining cell fate. Cells of the immune system are under enormous proliferative demand, stressing telomeric intactness. Lymphocytes are capable of upregulating telomerase, an enzyme that can elongate telomeric sequences and, thus, prolong cellular lifespan. Therefore, telomere dynamics are critical in preserving immune function and have become a focus for studies of immunosenescence and autoimmunity. In this review, we describe the role of telomeric nucleoproteins in shaping telomere architecture and in suppressing DNA damage responses. We summarize new insights into the regulation of telomerase activity, hereditary disorders associated with telomere dysfunction, the role of telomere loss in immune aging, and the impact of telomere dysfunction in chronic inflammatory disease. PMID- 20016141 TI - Three hemoglobin variants caused by a single alpha-chain gene mutation in a Chinese family. PMID- 20016142 TI - Differential effects of paricalcitol and calcitriol on intestinal calcium absorption in hemodialysis patients. AB - BACKGROUND/AIMS: Increased parathyroid activity associated with chronic kidney disease is often managed with calcitriol, which can elevate serum calcium (Ca) by increasing bone resorption and intestinal absorption, whereas paricalcitol promotes less bone resorption. This study compared intestinal Ca absorption in hemodialysis patients treated with calcitriol versus paricalcitol (dose ratio 1:3). METHODS: Patients (n = 22) aged > or =20 years, on maintenance hemodialysis for > or =2 months with intact parathyroid hormone (iPTH) levels of >200 pg/ml were enrolled in a single-center, double-blind, active-controlled, randomized, crossover trial. Mean fractional intestinal Ca absorption (+/-SE) was measured by the single-tracer method ((42)Ca) and evaluated with an analysis of variance crossover model. RESULTS: Mean fractional intestinal Ca absorption was significantly lower after paricalcitol (0.135 +/- 0.006) versus calcitriol treatment (0.158 +/- 0.006, p = 0.022), a 0.023 difference in absolute Ca absorption fraction. Overall Ca absorption was low in the study population, indicating that regulation of Ca absorption may be dysfunctional. There were no significant differences in serum PTH, Ca, phosphorus (P), or Ca x P. CONCLUSION: Overall, paricalcitol-treated patients absorbed approximately 14% less Ca compared with calcitriol-treated patients with similar effects on PTH. In hemodialysis patients, paricalcitol may provide a benefit by lowering the Ca available for removal by dialysis and/or for deposit in bone or soft tissues. PMID- 20016143 TI - Alterations in the renin-angiotensin system in a rat model of human preeclampsia. AB - BACKGROUND/AIMS: Preeclampsia is a hypertensive disorder unique to pregnancy in which elevated levels of marinobufagenin (MBG) have been reported. The renin angiotensin system (RAS) may also play a role in the pathogenesis of preeclampsia. The aim of our study was to evaluate the status of the RAS in a rat model of preeclampsia characterized by hypertension, proteinuria, excessive weight gain and intrauterine growth restriction. METHODS: We evaluated the components of the RAS in 5 groups of animals: nonpregnant control; normal pregnant (NP); pregnant rats which received injections of desoxycorticosterone acetate and 0.9% saline as their drinking water (PDS); normal pregnant rats injected with MBG (NPM), and PDS rats to which resibufogenin (RBG) had been administered (PDSR). RBG is an antagonist of MBG differing in structure from MBG only in the absence of a hydroxyl group in the beta-5 position. RESULTS: Plasma levels of active renin, renin and Ang II were significantly lower in PDS and NPM compared to NP and PDSR rats (p < 0.05). However, placental levels of these components were increased significantly in PDS and NPM compared to NP and PDSR rats (p < 0.05). Placental AT(1) receptor expression was significantly higher in PDS and NPM compared to NP and PDSR rats (p < 0.05). CONCLUSIONS: (1) The peripheral RAS is downregulated, and the uteroplacental RAS is upregulated in this rat model of preeclampsia; (2) MBG is involved in the causation of these alterations, and (3) RBG prevents these changes. PMID- 20016144 TI - N-acetylcysteine amide protects renal proximal tubular epithelial cells against iohexol-induced apoptosis by blocking p38 MAPK and iNOS signaling. AB - BACKGROUND: The pathogenesis of contrast-induced nephropathy (CIN) is still poorly understood and apoptosis via oxidative stress has been proposed as one possible mechanism. We therefore studied the apoptotic signaling mechanism in CIN and also tested whether the new antioxidant N-acetylcysteine amide (NACA) could prevent CIN. METHODS: LLC-PK1 cells were exposed to a widely used contrast agent, iohexol (IH). Cytotoxicity was assessed with morphology and 3-(4,5 dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. Cell death was analyzed by the DNA content analysis and PARP cleavage. Protein expression was assessed with Western blotting. RESULTS: We observed cell death with apoptotic features in a dose- and time-dependent manner. Initiation of IH-induced apoptosis was mediated by upregulation of Bax and downregulation of Bcl-2 and Mcl-1, which was preceded by p38 MAPK activation and iNOS induction. Inhibitors of p38 MAPK and iNOS partially abolished IH-induced apoptosis. Furthermore, we found pretreatment with NACA partially protected cells from IH-induced death by reverting the expression of Bcl-2, Mc1-1 and Bax expression through inhibition of p38 MAPK and iNOS pathway. CONCLUSIONS: This study demonstrates that apoptosis occurs during CIN. Apoptosis is associated with activations of p38 MAPK and iNOS. Pretreatment with the antioxidant NACA could prevent IH-induced cell death by blocking the p38 MAPK/iNOS signaling pathway. PMID- 20016145 TI - Effect of food restriction on energy expenditure of monosodium glutamate-induced obese rats. AB - The neonatal administration of monosodium glutamate (MSG) to rodents leads to obesity in the adult animal, characterized by increased fat storages. Chronic food restriction is known to induce reduction in body energy expenditure, as an adaptive mechanism to save energy. Our purpose was to examine whether obesity can alter the mechanism of energy conservation in food-restricted animals. Newborn female Wistar rats were injected either MSG (obese) or saline (control). At the age of 90 days, the animals were fed daily ad libitum (control and MSG) or restricted (50%) (control-restricted and MSG-restricted). After 30 days the animals were sacrificed and the energy balance was determined by calorimetric analysis. Some parameters of energy balance and body composition were affected by MSG treatment as well as food restriction. The percent reduction of the energy expenditure and fat content in MSG-restricted animals was lower than control restricted animals, when compared with their respective ad libitum groups. These results indicate that all food-restricted animals were able to develop the mechanism of energy conservation, regardless of the obesity, but it was less efficient in MSG-obese animals. PMID- 20016146 TI - Ultrasound measurement of total body fat in obese adolescents. AB - BACKGROUND/AIMS: To compare body fat (BF) measurements obtained with a new ultrasound method with those assessed by dual-energy X-ray absorptiometry (DEXA) in obese adolescents. METHODS: In 94 adolescents (57 females and 37 males) aged 12-19 years and body mass index (BMI) exceeding 30 kg.m(-2), the z-score BMI for age was 6.7 (adolescent girls) and 6.6 (adolescent boys) >97th percentile. BF was measured using DEXA and a method based on ultrasound measurements, body weight, height, abdominal circumference and mid-thigh circumference. RESULTS: Obesity class I was noted in 39%, II in 28% and III in 33% of the patients. BF by ultrasound correlated closely with BF by DEXA, in both females (r = 0.958) and males (r = 0.981), with standard errors of the estimates (SEE) being 2.9 and 2.5 kg, respectively. The ultrasound method was more accurate than the skinfold technique (n = 24; SEE: 2.2 vs. 6.5 kg, respectively). In 13 adolescents who had marked weight loss after 6 months of treatment, the decrease in DEXA-measured BF correlated closely with the decrease in ultrasound-measured BF (r = 0.95). CONCLUSIONS: Our innovative portable ultrasound technique has advantages in terms of reliability, reproducibility, accuracy and costs for screening and monitoring obese adolescents. A patent application has been submitted. Our method should prove valuable for epidemiological studies. PMID- 20016147 TI - Ameliorative effects of stabilized rice bran on type 2 diabetes patients. AB - BACKGROUND/AIM: While it has been demonstrated that rice bran might lower the cholesterol level in hypercholesterolemic individuals, its effects on the levels of adiponectin and glycated hemoglobin (HbA1c) in patients with type 2 diabetes mellitus remain unknown. METHODS: Twenty-eight volunteers with type 2 diabetes were randomly divided into 2 groups, one of which received a dietary supplement of 20 g of stabilized rice bran and the other placebo once daily for 12 weeks. Parameters such as the level of HbA1c, glucose, insulin, homeostasis model assessment for estimation of relative insulin resistance, high-density and low density lipoprotein (LDL) cholesterol and adiponectin were evaluated. RESULTS: At the end of the study period, postprandial glucose and the area under the glucose curve of the rice bran group were significantly lower than baseline levels by 14.4 and 15.7%, respectively. Compared to baseline, the HbA1c values in the rice bran group were also significantly lower. Serum total cholesterol and LDL cholesterol concentrations in the rice bran group were 9.2 and 13.7% lower, respectively, than in the placebo group. The plasma free fatty acid and adiponectin concentrations were 20% lower and 40% higher in the rice bran group compared to the placebo group. CONCLUSIONS: This study demonstrated that stabilized rice bran can lower the level of HbA1c and blood lipids and increase blood adiponectin concentrations in type 2 diabetic subjects. In light of this, we conclude that stabilized rice bran may represent an important functional nutrient to ameliorate lipid and glycemic anomalies in type 2 diabetic subjects. PMID- 20016148 TI - Blood volume- and blood temperature-controlled hemodialysis in critically ill patients: a 6-month, case-matched, open-label study. AB - BACKGROUND/AIMS: Online monitoring devices for intermittent hemodialysis have been developed to improve intradialytic cardiovascular stability in chronic dialysis patients. We propose to test blood volume and blood temperature biofeedback control systems in critically ill patients with acute kidney injury (AKI). METHODS: Twenty patients were prospectively dialyzed with machines equipped with online monitoring new devices and compared with 42 historical controls. In addition to feasibility and safety appraisals, efficacy was assessed by the intradialytic hypotension rate. RESULTS: During the study period, controlled temperature was assessed by a mean Delta(max) body temperature of -0.3 +/- 0.2 degrees C, and all but 7% of dialysis sessions achieved relative blood volume-guided ultrafiltration profiling without increased dialysis-related complications. This was associated with decreased intradialytic hypotension rate. CONCLUSION: The combination of active controlled body temperature and profiled ultrafiltration by online monitoring systems is feasible and safe in critically ill AKI patients, and suggests possibilities for improvement in intradialytic hemodynamic stability. PMID- 20016149 TI - Endothelial bioreactor ameliorates endotoxemia sepsis in swine. AB - INTRODUCTION: Severe sepsis involves a complex response including the activation of lots of cells, inflammatory mediators, and the hemostatic system. Central to this process is an alteration of endothelial cell function. Therefore, we investigated whether an endothelial bioreactor (EBR) would provide a new therapeutic approach to this clinical disorder. METHODS: EBR was constructed using a cartridge which contained with nonwoven polytetrafluoroethylene seeded with porcine iliac artery endothelial cell (PIEC). Pigs were intravenously administered with 0.25 mg/kg lipopolysaccharide and immediately placed in an extracorporeal circuit with EBR or sham controls. RESULTS: Compared with the sham group, EBR therapy resulted in significantly higher mean arterial blood pressure and significantly lower plasma von Willebrand factor, endothelin-1 and scores of lung injury. These alterations were associated with a significant survival advantage in the EBR group. CONCLUSIONS: Timely intervention with EC therapy in a tissue-engineered bioreactor may improve cardiovascular performance and alter the natural history of endotoxemia sepsis. PMID- 20016150 TI - Middle-molecule clearance at 20 and 35 ml/kg/h in continuous venovenous hemodiafiltration. AB - BACKGROUND: Of 5 clinical trials testing dose response of continuous renal replacement therapy (CRRT) in acute kidney injury, 2 showed a benefit, 2 showed none, and 1 appeared equivocal. However, blood-membrane interactions may dominate macromolecule transport in continuous venovenous hemodiafiltration, reducing the impact of dose adjustment. The dosing arms in the Acute Renal Failure Trial Network (ATN) study may have delivered similar clearances for middle molecules. METHODS: We simulated the 2 CRRT doses in the ATN study using a synthetic polydisperse macromolecular probe in bovine blood. Clearance of tracers between 10 and 100 kDa molecular weight was measured during 6 h of therapy. RESULTS: Middle-molecule clearance differed by less than 2 ml/min between the 2 dosing arms. CONCLUSION: The CRRT prescription used in the ATN study appears to have achieved dose separation for small molecules while holding middle-molecule clearance nearly constant. This may explain the outcome difference between the ATN study and earlier studies, and suggests subsequent trial designs. PMID- 20016151 TI - Molecular and evolutionary cytogenetics of domestic animals. Preface. PMID- 20016152 TI - Parvathi K. Basrur. PMID- 20016153 TI - Satellite DNA in the karyotype evolution of domestic animals--clinical considerations. AB - Eukaryotic genomes contain far more DNA than needed for coding proteins. Some of these additional DNA sequences comprise non-coding repetitive DNA sequences, mostly satellite DNAs and also transposable elements usually located at the heterochromatic regions of chromosomes. Satellite DNAs consist of tandemly repeated DNA sequences inhabiting the mammalian genome, typically organized in long arrays of hundreds or thousands of copies. Different important functions have been ascribed to satellite DNA, from the imperative centromeric function in mitosis and meiosis to the recent discovery of its involvement in regulatory functions via satellite transcripts. Moreover, satellite DNAs, among other repetitive sequences, are believed to be the 'engine' triggering mammalian genome evolution. Repetitive DNAs are, most likely, the genetic factors responsible for promoting genomic plasticity and therefore higher rates of chromosome mutation. Furthermore, constitutive heterochromatin regions are thought to be 'hotspots' for structural chromosome rearrangements. A considerable collection of evidences places these sequences in the landscape of mammalian evolution. However, the mechanisms that could explain how this alliance between chromosome evolution and satellite DNA is made are still enigmatic and subject of debate. Throughout the mammalian taxa, different patterns of chromosome evolution have been widely registered from heterochromatin additions/eliminations, Robertsonian translocations, whole-arm reciprocal translocations to tandem translocations; the fact is genome's repetitive fraction is playing a central role in mammalian genome structuring. Throughout this review we will focus on the evidences that associate satellite DNAs and constitutive heterochromatin to the process of chromosome evolution and consequently to domestic species genome's remodeling. PMID- 20016154 TI - Contribution of radiation hybrids to genome mapping in domestic animals. AB - Radiation hybrid mapping has emerged in the end of the 1990 s as a successful and complementary approach to map genomes, essentially because of its ability to bridge the gaps between genetic and clone-based physical maps, but also using comparative mapping approaches, between 'gene-rich' and 'gene-poor' maps. Since its early development in human, radiation hybrid mapping played a pivotal role in the process of mapping animal genomes, especially mammalian ones. We review here all the different steps involved in radiation hybrid mapping from the constitution of panels to the construction of maps. A description of its contribution to whole genome maps with a special emphasis on domestic animals will also be presented. Finally, current applications of radiation hybrid mapping in the context of whole genome assemblies will be described. PMID- 20016155 TI - Fluorescence in situ hybridization applied to domestic animal cytogenetics. AB - The aim of this article is not to present an exhaustive review of molecular cytogenetics applications in domestic animal species, but more to illustrate the considerable contribution of these approaches in diagnostics and research in economically important species. A short presentation of the main applications of molecular cytogenetics in humans points out the domains in which it has become an essential tool and underlines the specificities attached to this species in comparison to farm animals. This article is devoted to outlining the current resources available in domestic species and to some examples of fluorescence in situ hybridization applications in the cattle, pig, horse and avian species. From a clinical point of view, these examples illustrate the advantages of FISH for the study of chromosomal abnormalities (identification, characterization and estimation of their effects). Other applications of molecular cytogenetics are also illustrated, particularly ZOO-FISH, an approach which allows the determination of chromosome homologies between species. Finally, a specific emphasis was placed on the usefulness of molecular cytogenetics for the analysis of species such as poultry, which harbour a complex karyotype. PMID- 20016156 TI - Chromosome evolution in domestic bovids as revealed by chromosome banding and FISH-mapping techniques. AB - The present review summarizes the basic cytogenetic information available pertaining to the most important Bovidae species, namely cattle, buffalo, sheep and goat, with the aim of tracing their evolutionary relationships and to provide - for the first time - the hypothetical ancestral karyotype of the Bovinae Caprinae subfamilies, also in relation to the other nondomestic species which are included in this important taxonomic family. Evolution of the Bovinae-Caprinae autosomes and gonosomes is discussed on the basis of the most recent advances in chromosome banding, linkage studies, FISH-mapping and molecular information. PMID- 20016158 TI - Molecular cytogenetics and comparative mapping in goats (Capra hircus, 2n = 60). AB - Few goat genome analysis projects have been developed in the last 10 years. The aim of this review was to compile and update all available cytogenetic mapping data, according to the last goat chromosome nomenclature, as well as human and cattle whole genome sequences. In particular, human regions homologous to most of the FISH-mapped microsatellites were identified in silico. This new goat cytogenetic map made it possible to refine delineation of conserved segments relative to the human and cattle genomic sequence. These improvements did not lead to detection of major new rearrangements within ruminants but confirmed the good conservation of synteny and the numerous intrachromosomal rearrangements observed between goats and humans. PMID- 20016157 TI - Molecular cytogenetics and gene mapping in sheep (Ovis aries, 2n = 54). AB - The development of a completely annotated sheep genome sequence is a key need for understanding the phylogenetic relationships and genetic diversity among the many different sheep breeds worldwide and for identifying genes controlling economically and physiologically important traits. The ovine genome sequence assembly will be crucial for developing optimized breeding programs based on highly productive, healthy sheep phenotypes that are adapted to modern breeding and production conditions. Scientists and breeders around the globe have been contributing to this goal by generating genomic and cDNA libraries, performing genome-wide and trait-associated analyses of polymorphism, expression analysis, genome sequencing, and by developing virtual and physical comparative maps. The International Sheep Genomics Consortium (ISGC), an informal network of sheep genomics researchers, is playing a major role in coordinating many of these activities. In addition to serving as an essential tool for monitoring chromosome abnormalities in specific sheep populations, ovine molecular cytogenetics provides physical anchors which link and order genome regions, such as sequence contigs, genes and polymorphic DNA markers to ovine chromosomes. Likewise, molecular cytogenetics can contribute to the process of defining evolutionary breakpoints between related species. The selective expansion of the sheep cytogenetic map, using loci to connect maps and identify chromosome bands, can substantially contribute to improving the quality of the annotated sheep genome sequence and will also accelerate its assembly. Furthermore, identifying major morphological chromosome anomalies and micro-rearrangements, such as gene duplications or deletions, that might occur between different sheep breeds and other Ovis species will also be important to understand the diversity of sheep chromosome structure and its implications for cross-breeding. To date, 566 loci have been assigned to specific chromosome regions in sheep and the new cytogenetic map is presented as part of this review. This review will also summarize the current cytogenomic status of the sheep genome, describe current activities in the sheep cytogenomics research sector, and will discuss the cytogenomics data in context with other major sheep genomics projects. PMID- 20016159 TI - Comparative genomics of 3 farm canids in relation to the dog. AB - There are 3 canids besides the dog (Canis familiaris): the red fox (Vulpes vulpes), arctic fox (Alopex lagopus) and Chinese raccoon dog (Nyctereutes procyonoides procyonoides), which have been extensively studied with the use of cytogenetic and molecular genetics techniques. These 3 species are considered as farm fur-bearing animals. In addition, they are also useful models in comparative genomic studies of the canids. In this review genome organization, karyotype evolution, comparative marker maps, DNA polymorphism and similarity of selected gene sequences of the 3 farm species are discussed in relation to the dog. Also the nature and variability of the B chromosomes, present in the red fox and the Chinese raccoon dog, were considered. These comparative analyses showed that among the studied canids the Chinese raccoon dog is phylogenetically the closest species to the dog. On the other hand, the most advanced linkage and cytogenetic marker maps of the red fox genome facilitate genome scanning studies with the aim to search for chromosome locations of QTL regions for behavior and production traits. PMID- 20016160 TI - Fragile sites in domestic animal chromosomes: molecular insights and challenges. AB - Fragile sites are intriguing cytogenetic phenomena that have been extensively investigated in human and laboratory animal chromosomes over the past 40 years, but domestic animal species have been studied sporadically. Interest in the field has been recently renewed as increasing numbers of fragile site regions are cloned and characterized at the molecular level. Despite much effort, mechanisms by which specific DNA sequence confers fragility to a chromosome region remain unclear. Recent advancements in sequencing and mapping of domestic animal genomes provide tools for molecular characterization of fragile sites in animal chromosomes. Future comparative efforts may contribute insight into both the mechanisms that underlie chromosome fragility, and forces that drive rearrangements observed throughout evolution, and somatic changes that can result in disease or impair fertility. PMID- 20016161 TI - Classical and molecular cytogenetics of disorders of sex development in domestic animals. AB - The association of abnormal chromosome constitutions and disorders of sex development in domestic animals has been recorded since the beginnings of conventional cytogenetic analysis. Deviated karyotypes consisting of abnormal sex chromosome sets (e.g. aneuploidy) and/or the coexistence of cells with different sex chromosome constitutions (e.g. mosaicism or chimerism) in an individual seem to be the main causes of anomalies of sex determination and sex differentiation. Molecular cytogenetics and genetics have increased our understanding of these pathologies, where human and mouse models have provided a substantial amount of knowledge, leading to the discovery of a number of genes implicated in mammalian sex determination and differentiation. Additionally, other genes, which appeared to be involved in ovary differentiation, have been found by investigations in domestic species such as the goat. In this paper, we present an overview of the biology of mammalian sex development as a scientific background for better understanding the body of knowledge of the clinical cytogenetics of disorders of sex development in domestic animals. An attempt to summarize of what has been described in that particular subject of veterinary medicine for each of the main mammalian domestic species is presented here. PMID- 20016162 TI - Comparative RH maps of the river buffalo and bovine Y chromosomes. AB - Radiation hybrid maps were constructed for river buffalo and cattle Y chromosomes. A total of 41 cattle-derived Y-chromosome molecular markers were selected and tested with 2 previously described 5,000-rad whole-genome radiation hybrid (RH) panels (river buffalo - BBURH(5000) and cattle - BTARH(5000)) for generation of maps. Among the initial 41 selected markers, a subset of 26 markers generated PCR products suitable for scoring with the BBURH(5000) panel. Of these, 19 markers (73%) were distributed in 1 linkage group spanning 341.3 cR. Retention frequencies (RF) for individual markers ranged from 17.8% for SMCY to 56.7% for BTY1, with an average RF of 37.6%. From the selected markers, 37 generated reliable scores using the BTARH(5000) panel. The newly constructed BTAY RH map contains 28 markers distributed within 1 linkage group. Twenty-four of these markers had been previously mapped on BTAY using a 7,000-rad cattle-hamster WG-RH panel and 4 markers were mapped for the first time (ZFY, SeqRep, RepSeqS4 and BTY1). The length of the BTAY RH map was estimated to be 602.4 cR. Retention frequencies for individual mapped markers ranged from 10% (INRA126) to 63.3% (SeqRep), with an average RF of 35.3%. RH marker positions along the Y chromosome were compared between BBUY and BTAY, which revealed differences in the order of some of the markers. The BBUY pseudoautosomal region (PAR) is delineated by 3 BTAY PAR markers (MAF45, TGLA325 and UMN2008). These markers are telomeric in both species but are not found in the same order. Here we have demonstrated the effective use of bovine Y chromosome markers for the development of the first BBUY RH map. Likewise, these set of markers can be used for comparative assessment of Y chromosomes in other members of the Bovidae family. PMID- 20016163 TI - Characterization of the bovine pseudoautosomal region and comparison with sheep, goat, and other mammalian pseudoautosomal regions. AB - The pseudoautosomal region (PAR) is a small region of sequence homology between mammalian X and Y chromosomes and is needed for sex chromosome segregation in male meiosis. The region, though studied as yet in only a few species, shows considerable variation in size and gene content. We have constructed a medium density gene map for the cattle PAR and the adjacent X-specific region by isolating and mapping 18 BAC clones which contain 20 PAR- and 5 X-specific genes. One BAC clone containing TBL1XY and GPR143 spanned the recently demarcated bovine pseudoautosomal boundary (PAB). Comparing the gene map of cattle PAR with the high-resolution maps of human, horse, and dog PAR allowed to estimate that the size of cattle PAR is approximately 5-9 Mb. BAC end sequence analysis showed that there is a gradient of decreasing GC content from PARter towards the PAB which is consistent with findings in human, mouse, and horse. The 20 PAR- and 5 X-specific cattle genes were mapped also in goat and sheep, showing that PAR in the 3 species is similar in size, gene content, and gene order. For the first time the PAB was determined in goat sex chromosomes. Comparison of cattle, goat, and sheep PAR with homologous regions on human and horse X chromosomes showed a high degree of linkage conservation between all species. However, the most terminal human, horse, and dog PAR gene, PLCXD1, is X-specific in ruminants. Since the human/horse linkage group containing PLCXD1 is of ancestral origin, the location of PLCXD1 can be considered as a de novo event in ruminant sex chromosome evolution. The gene map of the cattle PAR adds to our knowledge about the comparative organization and evolution of the eutherian PAR and aids the sequencing, sequence assembly, and annotation of the terminal region of BTAXq. PMID- 20016164 TI - Polymorphic heterochromatic segments in Japanese quail microchromosomes. AB - Using highly extended lampbrush chromosomes from diplotene oocytes, we have examined the distribution of heterochromatin protein 1 beta (HP1 beta) and histone H3 modifications on chicken (Gallus gallus) and Japanese quail (Coturnix japonica) (2n = 78) microchromosomes. Acrocentric microchromosomes of chicken and submetacentric microchromosomes of quail differ in several morphological features. In addition to pericentromeric and subtelomeric blocks of constitutive heterochromatin, which are enriched in HP1 beta protein and repressive histone modifications, not completely condensed but heterochromatic segments were found to be an attribute of the short arms of submetacentric microchromosomes in Japanese quail. These heterochromatic regions are variable in length and do not form chiasmata in female germ cells. Dissimilarity in the centromere positions in chicken and Japanese quail microchromosomes is proposed to be due to the accumulation of repetitive sequences on the short arms of quail microchromosomes. Transcriptional activation of polymorphic heterochromatic segments of quail microchromosomes during the lampbrush stage is demonstrated. PMID- 20016165 TI - An appraisal of nuclear organisation in interphase embryonic fibroblasts of chicken, turkey and duck. AB - Determining the nuclear 'addresses' of chromosome territories is a well documented means of assaying for nuclear organisation in many cell types and species. Data in avian species are however limited at best, despite the pivotal role played by birds (particularly chickens) in agriculture, and as model organisms in developmental biology. That is, studies have hitherto focussed mostly on mammals (especially humans) and have demonstrated the importance of chromosome territory positioning in embryology, disease and evolution. Thus a detailed study of nuclear organisation in many species, many cell types and many developmental stages in birds is warranted, however, before this is achieved, 'baseline' needs to be established to determine precisely the relative locations of chromosome territories in at least 1 cell type of at least 1 bird. With this in mind we hybridised FISH probes from chicken chromosomes 1-28 to embryonic fibroblast nuclei, determining nuclear addresses using a newly developed plug-in to the image analysis package ImageJ. In our experience, evenly spaced representative BAC clones yielded more consistent results than hybridisation of chromosome paints. Results suggested that chromosome territory distribution best fitted a chromosome size-based (rather than gene density-based) pattern. Identical BAC clones were then hybridised to turkey and duck in a comparative genomic strategy. Observations were consistent with those seen in chicken (although, less well-defined in duck), providing preliminary evidence of conservation throughout evolution. PMID- 20016166 TI - Phylogeny of horse chromosome 5q in the genus Equus and centromere repositioning. AB - Horses, asses and zebras belong to the genus Equus and are the only extant species of the family Equidae in the order Perissodactyla. In a previous work we demonstrated that a key factor in the rapid karyotypic evolution of this genus was evolutionary centromere repositioning, that is, the shift of the centromeric function to a new position without alteration of the order of markers along the chromosome. In search of previously undiscovered evolutionarily new centromeres, we traced the phylogeny of horse chromosome 5, analyzing the order of BAC markers, derived from a horse genomic library, in 7 Equus species (E. caballus, E. hemionus onager, E. kiang, E. asinus, E. grevyi, E. burchelli and E. zebra hartmannae). This analysis showed that repositioned centromeres are present in E. asinus (domestic donkey, EAS) chromosome 16 and in E. burchelli (Burchell's zebra, EBU) chromosome 17, confirming that centromere repositioning is a strikingly frequent phenomenon in this genus. The observation that the neocentromeres in EAS16 and EBU17 are in the same chromosomal position suggests that they may derive from the same event and therefore, E. asinus and E. burchelli may be more closely related than previously proposed; alternatively, 2 centromere repositioning events, involving the same chromosomal region, may have occurred independently in different lineages, pointing to the possible existence of hot spots for neocentromere formation. Our comparative analysis also showed that, while E. caballus chromosome 5 seems to represent the ancestral configuration, centric fission followed by independent fusion events gave rise to 3 different submetacentric chromosomes in other Equus lineages. PMID- 20016167 TI - A comparison of coding sequence and cytogenetic localization of the myostatin gene in the dog, red fox, arctic fox and Chinese raccoon dog. AB - The gene encoding myostatin (MSTN), due to its crucial function for growth of skeletal muscle mass, is an important candidate for muscularity. In this study we analyzed the nucleotide sequence and FISH localization of this gene in 4 canids, including 3 farm species. The nucleotide sequence of the MSTN coding fragment turned out to be highly conserved, since its identity among the studied species was very high and varied between 99.4 and 99.7%. Only 1, widely spread, silent single nucleotide polymorphism (SNP) was found in exon 1 of the Chinese raccoon dog. The MSTN gene was localized close to the centromere in one-armed chromosomes of the dog (37q11) and bi-armed chromosomes of the red fox (16p11) and arctic fox (10q11), with an exception of the Chinese raccoon dog chromosome (2q14-q21). This chromosome is orthologous to 3 canine chromosomes and thus the MSTN was found more interstitially. Our results are in agreement with the hypothesis that karyotypes of the canids evolved mainly through centric fusion/fission events, while tandem fusions occurred rarely. PMID- 20016168 TI - Comparative cytogenetic and molecular studies of DM domain genes in pig and cattle. AB - Despite great differences in the biology of sex determination, the DM domain genes are thought to regulate sexual development in both invertebrates and vertebrates. Although they seem to represent the first known example of the conservation of metazoan sexual pathways, the structure-function relationships of this gene family have not yet been clearly determined. Our previous and current studies on the DMRT (Doublesex and mab-3 related transcription factor) gene cluster, DMRT1, DMRT2, DMRT3, in pig and cattle, have revealed conserved linkage to bovine (BTA8q17) and swine (SSC1q21) autosomes. However, the DMRT cluster seems to harbour the genes in different orders in both cattle (Entrez database) and pig (IMpRH map), compared to other vertebrates investigated so far. Moreover, we found that expression of DMRT1 (considered as the testicular regulator so far) is not restricted to gonads in adult animals. Beside ovary and testis, DMRT1 expression was detectable in swine kidney and bovine liver, heart, skeletal muscle, kidney, lung, spleen and showed no differences between the sexes. Swine and bovine DMRT3 expression was strongly restricted to the adult testis, while DMRT2 homologues have a wider expression pattern, as occurs in other mammals. The significance of this expression pattern of DMRT genes in adult pig and cattle remains unclear. It might shed new light on mammalian DM domain genes evolution and livestock intersexuality. PMID- 20016169 TI - Comparative mapping and genomic annotation of the bovine oncosuppressor gene WWOX. AB - WWOX (WW domain-containing oxidoreductase) is the gene mapping at FRA16D HSA16q23.1, the second most active common fragile site in the human genome. In this study we characterized at a detailed molecular level WWOX in the bovine genome. First, we sequenced cDNA from various tissues and obtained evidence in support of a 9-exon structure for the gene, similar to the human gene. Then, we recovered BACs using exon tags and annotated the gene to a >1-Mb genomic region of BTA18 using the Btau 4.0 genome assembly as a reference, thus resolving an issue related to exon 9, which is not included in the genomic annotation of the gene in the Entrez database. Finally, BACs spanning WWOX were used as FISH probes to obtain comparative mapping of the gene in Bos taurus, Bubalus bubalis, Ovis aries and Capra hircus to BTA18q12.1, BBU18q13, OAR14q12.1 and CHI18q12.1, respectively. Our data show that the chromosomal location of WWOX is conserved between man and 4 major domesticated species. Moreover, the annotation of the bovine gene also suggests a highly conserved genomic arrangement, including number and size of introns. PMID- 20016170 TI - Analysis using sperm-FISH of a putative interchromosomal effect in boars carrying reciprocal translocations. AB - The occurrence of interchromosomal effects (ICE) in reciprocal translocation carriers still remains contradictory in the human literature. We used the pig as an animal model to investigate whether the structure of the reciprocal translocations as well as the size and/or type of the chromosomes not involved in the rearrangement may influence the occurrence and the extent of ICE. Analyses of chromosomal sperm content by fluorescence in situ hybridization (FISH) using whole-chromosome painting probes for 7 chromosomes (1, 10, 11, 13, 18, X and Y) were carried out on sperm samples of 2 boars with normal semen parameters carrying different balanced reciprocal translocations: 38, XY, t(3;15)(q27;q13) or 38, XY, t(12;14)(q13;q21). One fertile boar with normal karyotype was also studied as a control. Aneuploidy rates for the 7 chromosomes were estimated by scoring 10,000 to 20,000 spermatozoa for each probe combination. No significant ICE was found except for chromosome 1 in the case of the t(3;15) translocation. Even if statistically significant, this ICE remained very weak and should have very little impact on the reproductive performance of the carrier boar. The size and/or type of chromosomes not involved in the translocation do not seem to have a major influence on the occurrence of ICE. The structure of the translocation could play a role, but complementary studies should be carried out to confirm this assumption. PMID- 20016171 TI - Macrocephaly in bull spermatozoa is associated with nuclear vacuoles, diploidy and alteration of chromatin condensation. AB - Spermatozoa from 2 dairy AI (artificial insemination) bulls (A and B), identified by their abnormal spermiogram with cells depicting frequent macrocephaly, double tails and nuclear vacuoles, were case-investigated and compared to normal spermatozoa from a control AI sire (C). Head sizes were measured and morphological abnormalities scored using brightfield and differential interference contrast microscopy. The degree of sperm maturation and of resistance to acid-induced DNA denaturation in situ were determined after uploading of acridine orange using flow cytometry of 5,000 cells/sample. Nuclear fragmentation, i.e. the ratio of red to total (red + green) fluorescence, reached 7.1% and 31% in bulls A and B, compared to 2% in bull C. The proportion of immature spermatozoa, i.e. those with incomplete histone-protamine exchange and depicting higher green fluorescence compared to the main population of the control bull, reached 9.54% in A and 7.75% in B, compared to only 0.47% in the control. In the second part of this study the previously unknown chromosomal constitution of large-headed spermatozoa of bull A was investigated by fluorescence in situ hybridization using an X-Y painting probe set. The 7.5% XY bearing cells and the presence of diploid spermatozoa detected by flow cytometry indicate a meiotic arrest in the first division in bull A, becoming the first proven case of association of macrocephaly and M1 diploidy. The diverse approaches used for the investigation of spermatozoal DNA provide insights into the etiology of macrocephaly. PMID- 20016172 TI - Investigation of chromosome aneuploidies in early porcine embryos using comparative genomic hybridization. AB - Although numerical chromosome errors are known to be prevalent in early human embryos and are likely to be a considerable factor influencing the mortality of early embryos and implantation failure, in domestic animals data about the frequency and nature of errors is limited. The objectives of this study were to investigate the whole chromosome set of in vivo obtained early pig embryos, applying methods of whole genome amplification and comparative genomic hybridization (CGH) and to contribute to the comprehensive understanding of the topic. The embryos were collected from gilts 72 h after insemination. Further, they were lysed and underwent whole genome amplification by multiple displacement amplification. In a subsequent CGH, amplified DNA was compared to control DNA using different fluorescent labeling and hybridization to male pig mitoses. 11 (14.3%) of the 77 pig embryos examined were observed to be aneuploid. We found chromosome errors comprising loss/gain of one or a few chromosomes (10.4%) but also extensive chromosome imbalances (3.9%). Chromosomes 8, 11, 12, 13, 17, and X were most frequently involved in aneuploidies, when compared to chromosomes 2, 9, and 18, which were rarely involved in chromosome errors. PMID- 20016173 TI - X-Y sperm aneuploidy in 2 cattle (Bos taurus) breeds as determined by dual color fluorescent in situ hybridization (FISH). AB - The present study was undertaken to investigate aneuploidy rates in the sperm populations of 2 cattle (Bos taurus) breeds by using dual color fluorescent in situ hybridization (FISH) with Xcen and Y chromosome-specific painting probes, obtained by chromosome microdissection and DOP-PCR. Frozen semen from 10 Italian Friesian and 10 Italian Brown testing bulls was used for the investigation. For each bull, more than 5,000 sperm were analyzed, for a total of 52,586 and 51,342 sperm cells for the 2 breeds, respectively. The present study revealed - in both breeds - a preponderance of the Y-bearing sperm compared to the X-bearing sperm. Within each breed, a statistically significant variation in the various classes of aneuploidy (XX, YY and XY) was found: differences were found in the Friesian breed among the 3 diploidy classes, and in the Brown breed, among the 3 disomy classes (p < 0.05) as well as among the 3 diploidy classes (p < 0.01). However, the 2 breeds did not differ significantly in the overall mean rates of X-Y aneuploidy (disomy + diploidy) which amounts to 0.162% in the Italian Friesian and 0.142% in the Italian Brown. When meiosis I (MI) and II (MII) errors were compared, statistically significant differences (p < 0.01) were found in the disomy classes and in both breeds, whereas the differences between diploidy classes were not significant. Compared to humans, a lower level of aneuploidy has been found in the domestic species analyzed so far. The present study contributes to the establishment of a baseline level of aneuploidy in the sperm populations of 2 cattle breeds which could be used for monitoring future trends of reproductive health, especially in relation to environmental changes and mutagens. PMID- 20016174 TI - Doxorubicin cardiomyopathy. AB - Established doxorubicin cardiomyopathy is a lethal disease. When congestive heart failure develops, mortality is approximately 50%. Extensive research has been done to understand the mechanism and pathophysiology of doxorubicin cardiomyopathy, and considerable knowledge and experience has been gained. Unfortunately, no effective treatment for established doxorubicin cardiomyopathy is presently available. Extensive research has been done and is being done to discover preventive treatments. However an effective and clinically applicable preventive treatment is yet to be discovered. PMID- 20016175 TI - Six months of daily high-dose xylitol in high-risk schoolchildren: a randomized clinical trial on plaque pH and salivary mutans streptococci. AB - A randomized clinical trial was designed to evaluate the effect of daily high dose xylitol chewing gum on plaque pH and salivary mutans streptococci (MS) in a sample of schoolchildren at high risk of caries. The study was performed on 204 subjects (acceptance rate 88.3%). Inclusion criteria were: >1 and <4 carious lesions, and a salivary MS concentration >10(5) CFU/ml. Subjects were randomly assigned to the xylitol or control group. Study design included one examination at baseline (t(0)), one after 3 months of chewing (t(1)), one after 6 months of chewing (t(2)) and the last 3 months after the end of chewing period (t(3)). Plaque pH was assessed using the MicroTouch technique, following a sucrose challenge. The area under the curve (AUC(5.7) and AUC(6.2)) was recorded. Whole saliva was collected in sterile vials and MS CFU/ml were counted. Data were analysed using repeated-measures ANOVA. The main result was that plaque acidogenicity was reduced in both groups. The differences between treatments were statistically significant both for plaque pH and MS concentration; the interaction term for treatment and time was statistically significant (p < 0.01). At t(2), the xylitol group children with a salivary MS concentration >10(5) and those with < or =10(5) showed significantly lower AUC(5.7) and AUC(6.2) values than the control group. These results suggest that the long-term use of high-dose non-sucrose chewing gums had beneficial effects on plaque pH, and that this effect was statistically greater when using xylitol chewing gums, both on plaque pH and MS salivary concentration. PMID- 20016176 TI - Effect of various rinsing protocols after use of amine fluoride/stannous fluoride toothpaste on the bacterial composition of dental plaque. AB - This clinical study evaluated the effect of different oral hygiene protocols on the bacterial composition of dental plaque. After a 2-week period of using fluoride-free toothpaste, 30 participants followed three 1-week experimental protocols, each followed by 2-week fluoride-free washout periods in a randomized crossover examiner-blind controlled trial. The 1-week experimental protocols comprised the use of AmF/SnF(2) toothpaste twice daily, after which participants either (1) rinsed with tap water, (2) did not rinse but only spat out the toothpaste, or (3) rinsed with an AmF/SnF(2) mouthwash. In the fluoride-free washout periods, the participants brushed their teeth with fluoride-free toothpaste without further instructions. Six hours after the last brushing (+/- rinsing) of each period, buccal plaque samples in the upper molar region were taken. The microbiota composition of the plaque samples was analyzed by checkerboard DNA:DNA hybridization. A statistically significant reduction was found in the total amount of DNA of the 39 major plaque species measured, and in the proportions of some acid-producing bacterial strains after the period having used the AmF/SnF(2) toothpaste + AmF/SnF(2) mouthrinsing. The results indicate that using the AmF/SnF(2) toothpaste and rinse combination could result in plaque of lower cariogenicity. PMID- 20016177 TI - Association between caries experience and body mass index in 12-year-old French children. AB - The prevalence of overweight and obesity reached 19.7% in 12-year-old French children in the year 2005. Recently, nationwide programs have been broadly implemented in France to reduce the overconsumption of sugars, salt and fat. The aims of this study were to assess the distribution of body mass index (BMI) and D(3+4)MFT index in a sample of 12-year-old French children, and to compare several regression models in order to analyze the association between these two indices. A cross-sectional study was conducted in Montpellier, France, and the height, weight, D(3+4)MFT, sugar and soft drink consumption were recorded in a randomly selected sample of 835 schoolchildren. In order to analyze the association between BMI and DMFT, four models of regression were tested: logistic, Poisson, zero-inflated Poisson (ZIP) and zero-inflated negative binomial (ZINB). The mean BMI was 18.9 for the whole sample and the corresponding DMFT value was 1.47. The caries prevalence was 51.7%. The best fitted models for testing the association between BMI and DMFT were ZIP and ZINB models. They showed a significant association between DMFT and sugar consumption, but not with BMI. As a result of the best fitted models (ZIP and ZINB), where BMI was not statistically associated with DMFT, we conclude, within the limits of a cross sectional survey, that there is no association between these two variables. PMID- 20016178 TI - Comparison of cross-sectional hardness and transverse microradiography of artificial carious enamel lesions induced by different demineralising solutions and gels. AB - The aims of this study were: (1) to correlate surface (SH) and cross-sectional hardness (CSH) with microradiographic parameters of artificial enamel lesions; (2) to compare lesions prepared by different protocols. Fifty bovine enamel specimens were allocated by stratified randomisation according to their initial SH values to five groups and lesions produced by different methods: MC gel (methylcellulose gel/lactic acid, pH 4.6, 14 days); PA gel (polyacrylic acid/lactic acid/hydroxyapatite, pH 4.8, 16 h); MHDP (undersaturated lactate buffer/methyl diphosphonate, pH 5.0, 6 days); buffer (undersaturated acetate buffer/fluoride, pH 5.0, 16 h), and pH cycling (7 days). SH of the lesions (SH(1)) was measured. The specimens were longitudinally sectioned and transverse microradiography (TMR) and CSH measured at 10- to 220-microm depth from the surface. Overall, there was a medium correlation but non-linear and variable relationship between mineral content and radicalCSH. radicalSH(1) was weakly to moderately correlated with surface layer properties, weakly correlated with lesion depth but uncorrelated with integrated mineral loss. MHDP lesions showed the highest subsurface mineral loss, followed by pH cycling, buffer, PA gel and MC gel lesions. The conclusions were: (1) CSH, as an alternative to TMR, does not estimate mineral content very accurately, but gives information about mechanical properties of lesions; (2) SH should not be used to analyse lesions; (3) artificial caries lesions produced by the protocols differ, especially considering the method of analysis. PMID- 20016179 TI - Randomized equivalence trial of intensive and semiannual applications of fluoride varnish in the primary dentition. AB - For children in the primary dentition with high caries prevalence the standard semiannual application of fluoride varnish may not be successful in preventing tooth decay. Oftentimes this population is mobile and does not receive consistent preventive care. This trial tested whether an intensive fluoride 5% sodium varnish regimen (three applications/2 weeks) applied annually has an equivalent effect on caries progression in the primary dentition compared to single applications applied semiannually. This study was a randomized clinical trial with two treatment groups. All participants (n = 600; mean age +/- SD = 55.3 +/- 4.6 months) received three varnish applications (active varnish or placebo) at semiannual visits over 3 years. Once per year the intensive-treatment group received one set of three active treatments and three placebo treatments 6 months later, each time within 2 weeks. The standard group received one active and two placebo treatments every 6 months. Children were assessed clinically at baseline and 12, 24 and 36 months after the initiation of the study. The mean (SD) numbers of newly decayed primary tooth surfaces observed over 3 years were 9.8 (8.6) and 7.4 (7.7) in the intensive and standard groups, respectively. The adjusted rate ratio was 1.13 (95% CI = 0.94-1.37, p = 0.20). In conclusion, the trial failed to demonstrate clear evidence of a difference in efficacy. However, differences of up to 36% greater rates of caries in the intensive group could not be ruled out, thus equivalence of the treatments cannot be concluded. PMID- 20016180 TI - Cross-sectional hardness of enamel from human teeth at different posteruptive ages. AB - This study evaluated, by cross-sectional hardness (CSH), enamel maturation at different depths in sound human teeth at different posteruptive ages (12 per age group): unerupted (molars; control), 2-3 years (premolars), 4-10 years (premolars) and >10 years (molars). Cross-sectional Knoop hardness was measured at 10, 30, 50, 70, 90, 110, 220 and 330 mm depth from the surface. The data were subjected to two-way ANOVA followed by Bonferroni's pairwise tests (a = 0.05). The CSH significantly increased with posteruptive age, indicating that the time of exposure in the oral cavity might result in deep enamel maturation. PMID- 20016182 TI - Genetic variation in N-methyl-D-aspartate receptor subunit NR3A but not NR3B influences susceptibility to Alzheimer's disease. AB - BACKGROUND: The administration of memantine, an N-methyl-D-aspartate (NMDA) receptor antagonist, has clinically improved the cognitive function of patients with Alzheimer's disease (AD), indicating that a disturbance in glutamatergic transmission might be involved in a predisposition to developing the disease. AIM: The potential association of polymorphisms in NMDA receptor subunits NR3A and NR3B, encoded by the GRIN3A and GRIN3B genes, with AD was investigated. METHODS: We performed a case-control study. Two single nucleotide polymorphisms, 3104 G/A (rs10989563) and 3723 G/A (rs3739722), in the GRIN3A gene and 2 GRIN3B gene polymorphisms, 1210 C/T (rs4807399) and 1730 C/T (rs2240158), were studied. RESULTS: Upon genotyping of the exonic polymorphism in the GRIN3A gene, the G allele was present at a higher rate than the A allele at position 3723 in AD patients compared with normal groups (p < 0.05). Three haplotypes (designated Ht1 3) were identified from these 2 polymorphisms (3104 G/A and 3723 G/A), and the distribution of Ht2 (AG) differed between AD patients and controls (p < 0.05). Additionally, from the 2 GRIN3B gene variants 1210 C/T and 1730 C/T analyzed, no strong association with AD was observed. CONCLUSION: These observations suggest that the genetic variation of the NR3A, but not NR3B, subunit of the NMDA receptor may be a risk factor for AD pathogenesis among the Taiwanese population. PMID- 20016183 TI - Personality features in subjective cognitive impairment and mild cognitive impairment--early indicators of dementia? AB - OBJECTIVES: The purpose of the present study was to investigate patterns of personality in patients with subjective cognitive impairment (SCI) and mild cognitive impairment (MCI), compared to healthy controls. METHODS: We assessed24 patients with SCI, 35 patients with MCI and 26 healthy controls with the self report questionnaire Swedish Universities Scales of Personality measuring aspects of neuroticism/anxiety proneness, extraversion, and aggression-hostility. RESULTS: Patients with SCI and MCI showed significantly more Somatic Trait Anxiety, Psychic Trait Anxiety and Stress Susceptibility than healthy controls. Moreover, there was a significant increase in Detachment in patients with MCI and a significant decrease in Adventure Seeking in patients with SCI, relative to healthy controls. CONCLUSIONS: Patients with SCI and MCI presented specific patterns of personality alterations with higher scores in traits related to anxiety proneness and aggression-hostility and lower in traits of extraversion. In most subscales differences followed a sequential pattern with gradually increasing scores from healthy controls, to patients with SCI and further to MCI. The groups differed in amount and type of symptoms, suggesting that patterns of personality may be related to degree of cognitive impairment. PMID- 20016184 TI - Vitamin E use is associated with improved survival in an Alzheimer's disease cohort. AB - BACKGROUND: Vitamin E at a dose of 2,000 IU per day has been shown to delay Alzheimer's disease (AD) progression, but recent studies have questioned the safety of this dose level and the overall efficacy of vitamin E in AD treatment. METHODS: We analyzed the survival history of 847 probable or mixed AD patients followed in a research center between 1990 and the censoring date of December 31, 2004. Standard practice during this period was to recommend vitamin E at 1,000 IU twice daily to all patients. We used Cox proportional hazards modeling to assess the association of vitamin E alone, or in combination with a cholinesterase inhibitor (ChEI), with all-cause mortality, adjusting for important covariates. Approximately two thirds of the patients took vitamin E with a ChEI, 10% took vitamin E alone, and 15% took no antidementia drug. RESULTS: The adjusted hazard ratio (HR) associated with vitamin E (with or without a ChEI) was 0.71 (95% CI: 0.57-0.89; p = 0.003). Compared to the no drug treatment group, the HR for vitamin E alone or with another drug was 0.77 (95% CI: 0.60-1.0); the HR for ChEI use alone was 1.2 (95% CI: 0.87-1.60). CONCLUSION: The results do not support a concern over increased mortality with high-dose vitamin E supplementation. PMID- 20016185 TI - Single- and multiple-domain amnestic mild cognitive impairment: two sides of the same coin? AB - BACKGROUND: Amnestic mild cognitive impairment (aMCI) is considered a transition stage between normal aging and Alzheimer's disease (AD). Two main clinical subtypes of aMCI have been identified: (1) single-domain aMCI (aMCI-SD), with isolated episodic memory impairments, and (2) multiple-domain aMCI (aMCI-MD), with episodic memory impairments and deficits in one or more other cognitive domains. AIMS: To map the pattern of gray matter (GM) atrophy associated with aMCI-SD, aMCI-MD and mild AD. METHODS: A group of aMCI-SD, aMCI-MD characterized by executive function disorders, mild AD and cognitively unimpaired subjects underwent a comprehensive neuropsychological assessment and a high-definition MR brain scan. Voxel-based morphometry analysis was used to characterize the GM tissue loss in each patient group, and the common pattern of GM atrophy in aMCI SD and aMCI-MD. RESULTS: aMCI-SD and aMCI-MD are characterized by a common pattern of GM atrophy within the medial temporal cortex, predisposing to AD and correlating with the severity of verbal memory symptoms. Moreover, the pattern of GM atrophy observed in aMCI-SD, aMCI-MD and mild AD revealed that, from an anatomical point of view, these three clinical syndromes could represent three severity points along the continuum between normal aging and AD. PMID- 20016186 TI - Comparing outcomes of 'voluntary' and 'quasi-compulsory' treatment of substance dependence in Europe. AB - AIM: This study evaluates quasi-compulsory drug treatment (QCT) arrangements for substance-dependent offenders receiving treatment instead of imprisonment in comparison to voluntary treatment within five European countries. METHODS: Participants were interviewed with the European Addiction Severity Index, the ASI crime module, questions on perception of pressure and self-efficacy, and the Readiness-to-Change Questionnaire at treatment entry and after 6, 12, and 18 months. RESULTS: Reductions in substance use and crime as well as improvements in health and social integration were observed in QCT and voluntary treatment groups. After controlling for various factors, subjects in the QCT and the comparison group showed similar reductions in substance use and crime over time. Study retention was comparable in both groups. CONCLUSION: QCT is as effective as voluntary treatment provided in the same services in reducing substance use and crime. PMID- 20016187 TI - Fertility-sparing approach in a teenager with uterine tumor diagnosed as a sarcoma on imaging. AB - BACKGROUND: Uterine tumors are extremely rare in teenage patients. Although magnetic resonance imaging (MRI) can help distinguish between leiomyoma and sarcoma, it may also seriously mislead the diagnosis. CASE REPORT: An 18-year-old female patient presented with marked anemia, and a rapidly growing uterine tumor that strongly suggested uterine sarcoma on MRI. In order to avoid dispensable radical surgery, explorative laparotomy with tumor biopsy was performed after balancing risk/benefit ratio. Extemporaneous pathologic examination revealed a benign leiomyoma, and the patient subsequently underwent complete conservative myomectomy. CONCLUSION: Preserving fertility is an important issue in young patients with uterine tumors, and tumor biopsy may overcome MRI false positive results, as illustrated in this case. Conservative approach deserves further evaluation in this specific setting. PMID- 20016188 TI - Ultrasound experience substantially impacts on diagnostic performance and confidence when adnexal masses are classified using pattern recognition. AB - AIM: To determine how accurately and confidently examiners with different levels of ultrasound experience can classify adnexal masses as benign or malignant and suggest a specific histological diagnosis when evaluating ultrasound images using pattern recognition. METHODS: Ultrasound images of selected adnexal masses were evaluated by 3 expert sonologists, 2 senior and 4 junior trainees. They were instructed to classify the masses using pattern recognition as benign or malignant, to state the level of confidence with which this classification was made and to suggest a specific histological diagnosis. Sensitivity, specificity, accuracy and positive and negative likelihood ratios (LR+ and LR-) with regard to malignancy were calculated. The area under the receiver operating characteristic curve (AUC) of pattern recognition was calculated by using six levels of diagnostic confidence. RESULTS: 166 masses were examined, of which 42% were malignant. Sensitivity with regard to malignancy ranged from 80 to 86% for the experts, was 70 and 84% for the 2 senior trainees and ranged from 70 to 86% for the junior trainees. The specificity of the experts ranged from 79 to 91%, was 77 and 89% for the senior trainees and ranged from 59 to 83% for the junior trainees. The experts were uncertain about their diagnosis in 4-13% of the cases, the senior trainees in 15-20% and the junior trainees in 67-100% of the cases. The AUCs ranged from 0.861 to 0.922 for the experts, were 0.842 and 0.855 for the senior trainees, and ranged from 0.726 to 0.795 for the junior trainees. The experts suggested a correct specific histological diagnosis in 69-77% of the cases. All 6 trainees did so significantly less often (22-42% of the cases). CONCLUSION: Expert sonologists can accurately classify adnexal masses as benign or malignant and can successfully predict the specific histological diagnosis in many cases. Whilst less experienced operators perform reasonably well when predicting the benign or malignant nature of the mass, they do so with a very low level of diagnostic confidence and are unable to state the likely histology of a mass in most cases. PMID- 20016189 TI - Plasma adrenomedullin levels during multiple pregnancy. AB - BACKGROUND: To estimate the normal level of adrenomedullin (AM) during multiple pregnancy. METHODS: The study population consisted of 5 women with normal cycles, 130 women with normal pregnancy between 6 and 40 weeks of gestation, 93 women with twin pregnancy and 42 women with triplet pregnancy. RESULTS: Total AM concentration in the first trimester (13.7 +/- 0.58 fmol/ml), second trimester (26.8 +/- 1.13 fmol/ml) and third trimester (37.8 +/- 1.32 fmol/ml) in pregnant women was significantly higher than that in nonpregnant women (8.0 +/- 0.71 fmol/ml). In each trimester, the maternal plasma concentrations of triplet pregnant women were significantly higher than in twin and singleton-pregnant women. Umbilical venous AM levels (29.9 +/- 2.63 fmol/ml) were higher than umbilical arterial AM (20.2 +/- 2.04 fmol/ml). CONCLUSION: Since alterations in the AM concentration in maternal plasma may mediate compensatory vascular responses in the uterine circulation, it might be useful to know the normal level of AM during multiple pregnancy in order to detect abnormalities during multiple pregnancy. PMID- 20016190 TI - Timing of group B streptococcus screening in pregnancy: a systematic review. AB - BACKGROUND: Group B streptococcus (GBS) is an important cause of neonatal sepsis. Guidelines advise to collect cultures at 35-37 weeks' gestation and to administer intrapartum antibiotic prophylaxis in case of GBS-positive cultures, as well as in all preterm deliveries. Improved effectiveness of antenatal cultures might help to further decrease GBS early-onset disease. OBJECTIVE: To determine the best timing of antenatal cultures, which may help establish optimal prevention of perinatal GBS infection in both term and preterm neonates. METHODS: PubMed and EMBASE databases were searched for relevant articles published from 1966 to February 2009. Nine articles were included. Information about study features and predictive values of antenatal cultures were abstracted. RESULTS: Positive predictive values for antenatal GBS cultures ranged from 43 to 100% (mean 69%) and negative predictive values from 80 to 100% (mean 94%). GBS cultures collected in late pregnancy had high positive predictive values for colonization during delivery. The negative predictive value was high and relatively constant regardless of GA. CONCLUSIONS: This systematic review confirms recommendations to screen pregnant women for colonization of GBS at 35-37 weeks' gestation, but one should be aware of the limitations of screening, with 6% of GBS carriers remaining undetected in antenatal cultures. PMID- 20016191 TI - Topical 'Sydney' propolis protects against UV-radiation-induced inflammation, lipid peroxidation and immune suppression in mouse skin. AB - BACKGROUND: Propolis is a honeybee product that has been used in traditional medicine for antioxidant, immune-stimulating, anti-inflammatory and anti-cancer effects. Here, the potential of the topical application of a crude ethanolic extract of Sydney propolis to protect against UV-radiation-induced impairments associated with an increased risk of photocarcinogenesis has been tested in the hairless mouse. METHODS: Solutions providing between 10 and 200 mg/kg propolis were applied to the skin following UV irradiation. The inflammation from exposure to UV (290-400 nm) was quantitated by measurement of increased skinfold thickness; lipid peroxidation was assayed by the induction of thiobarbituric acid reactive species in the skin; immune function was measured by the contact hypersensitivity (CHS) reaction and supported by the changes in epidermal cytokine expression. RESULTS: Propolis protected significantly and dose dependently against both sunburn oedema and the suppression of CHS, and (at 100 mg/kg) against lipid peroxidation. The overexpression of IL-10 and the depletion of IL-12 characteristic of photoimmune suppression were markedly reduced by propolis. Further, the upregulation of IL-6 was decreased, and the associated induction of haem oxygenase was shown to play a role in propolis skin protection. CONCLUSIONS: Sydney propolis was able to effectively reduce cutaneous inflammation, immunosuppression and lipid peroxidation induced by UV exposure. It is concluded that Sydney propolis might have strong beneficial protective effects against photodamage and skin cancer development in humans. PMID- 20016193 TI - Association of plasma soluble CTLA-4 with lung function and gene polymorphism in Chinese asthmatic children. AB - BACKGROUND: Cytotoxic T lymphocyte antigen 4 (CTLA-4) is known to downregulate the T(H)2 immune response. Recent studies have suggested an association of CTLA-4 polymorphisms with allergic diseases. We investigated the effects of single nucleotide polymorphisms (SNPs) of CTLA-4 on asthma traits and plasma sCTLA-4 in 298 Chinese asthmatic children and 175 controls. METHODS: Plasma sCTLA-4, total and allergen-specific IgE concentrations were measured by enzyme immunoassay. Six SNPs, namely -1147CT, +49AG, CT60, JO31, JO30 and JO27_1, in CTLA-4 were genotyped by restriction fragment length polymorphism. RESULTS: Plasma sCTLA-4 was negatively associated with FEV(1)/FVC (r = -0.146, p = 0.036) among our asthmatic patients. Analysis of locus-locus interaction by generalized multifactor dimensionality reduction showed that -1147CT was the best model for plasma sCTLA-4 with a cross-validation consistency of 10 out of 10 and a prediction error of 40.9% (p < 0.001). Multivariate regression analysis confirmed the association between plasma sCTLA-4 concentration with -1147CT among the 6 SNPs tested (p = 0.002) after adjustment for gender and age. The plasma sCTLA-4 concentration was significantly lower in patients homozygous for the C allele than in T allele carriers (p = 0.001). There was also a significant association between the most common haplotypes with low sCTLA-4 in asthmatics. We could not find any significant association between plasma total IgE, atopy and lung function with the 6 SNPs after Bonferroni correction. CONCLUSIONS: Plasma sCTLA-4 is associated with lung function and -1147CT polymorphism in Chinese asthmatic children. This may help to identify CTLA-4 signaling as a potential therapeutic target in asthma. PMID- 20016192 TI - Intranasal CpG therapy attenuated experimental fungal asthma in a TLR9-dependent and -independent manner. AB - BACKGROUND: CpG administration abolishes airway inflammation and remodeling in acute models of allergic airway disease. METHODS: Herein, we investigated the therapeutic effect of CpG in a chronic fungal model of asthma. TLR9+/+ and TLR9-/ mice were sensitized to soluble Aspergillus fumigatus antigens and challenged with live A. fumigatus conidia. Mice were treated with intraperitoneal (IP) or intranasal (IN) CpG, or left untreated 14-28 days after conidium challenge. All features of allergic airway disease were attenuated in TLR9+/+ mice treated with IN CpG, including airway hyperresponsiveness (AHR), mucus production, and peribronchial fibrosis. RESULTS: TLR9-/- mice treated with IN CpG exhibited attenuated airway remodeling but not AHR. Whole-lung IL-12 levels were significantly elevated in both TLR9+/+ and TLR9-/- mice receiving IN CpG but not in either group receiving IP CpG. Whole-lung IL-10 levels were significantly elevated in IN CpG-treated TLR9+/+ mice but not in TLR9-/- mice receiving IN CpG. Increased whole-lung transcript and protein levels of the scavenger receptors SR A and MARCO were observed in TLR9-/- mice compared with TLR9+/+ mice, possibly accounting for the CpG responsiveness in the knockout group. CONCLUSIONS: Together, these data show that IN CpG has a therapeutic effect during established fungal asthma, which is TLR9 dependent and independent. PMID- 20016194 TI - Hydrolyzed Konjac glucomannan suppresses IgE production in mice B cells. AB - BACKGROUND: Oral administration of pulverized Konjac glucomannan (KGM) reduces increased plasma IgE and the amount of epsilon-germline transcript (epsilonGT) in the spleen, as well as preventing the development of dermatitis in mice. To elucidate the mechanism of action of pulverized KGM, we solubilized KGM and studied its effect on IgE in vitro and in vivo. METHODS: Solubilized KGM was prepared by acid hydrolysis, and we analyzed the effective molecular size for the suppression of IgE production and epsilonGT in vitro and the level of plasma IgE induced by immunization with ovalbumin in BALB/c mice. RESULTS: The production of IgE and epsilonGT in splenic cells, but not purified B cells, was inhibited by hydrolyzed KGM (KGM hydrolyzed with 0.25 N HCl; H-KGM) at the optimal size of between 10 and 500 kDa. However, no effect was observed when H-KGM was substituted with unhydrolyzed KGM in vitro. IgE production from purified B cells cocultured with purified monocytes, but not with purified T cells, was inhibited by H-KGM. The release of IFNgamma in cultures of monocytes but in purified B cells with or without T cells was enhanced in the presence of H-KGM. Injection of mice with H-KGM also suppressed the production of plasma IgE and IgG1 but not IgG2a in vivo. CONCLUSION: KGM at an optimal size prevents germline class switching and IgE production both in vitro and in vivo. H-KGM may be useful as a tool to study the mechanism of action of KGM and as a dietary supplement to prevent atopic diseases. PMID- 20016195 TI - A Toll-like receptor 2/6 agonist reduces allergic airway inflammation in chronic respiratory sensitisation to Timothy grass pollen antigens. AB - BACKGROUND: The hygiene hypothesis negatively correlates the microbial burden of the environment with the prevalence of T helper type 2 (Th2)-related disorders, e.g. allergy and asthma. This is explained by Th1 triggering through pathogen associated molecular patterns via Toll-like receptors (TLRs). In this study, the biological effects of a TLR2/6 agonist as a potential treatment of allergic inflammation are explored. METHODS: In a model of chronic allergic airway inflammation induced by intranasal administration of Timothy grass pollen allergen extract, early TLR agonism and/or interferon (IFN)-gamma administration was compared to the therapeutic and immune-modulating effects of dexamethasone with regard to the cellular inflammation and cytokine profiles. RESULTS: Eosinophilic inflammation was clearly reduced by TLR2/6 agonism. This effect was also seen without simultaneous administration of IFN-gamma. However, lymphocyte counts were not affected among the different treatment groups. More precise determination of the lymphocyte-mediated immune reaction showed that TLR2/6 agonism induced neither CD4+foxp3+ regulatory T cells in draining lymph nodes nor a pronounced Th1 immune response. In contrast, dexamethasone reduced both sensitisation as well as allergic inflammation and, in addition, CD11c+ antigen presenting cells in lymph nodes. Our data clearly point to the potential to rebalance Th2-skewed allergic immune responses by therapeutic TLR2/6 agonist administration. CONCLUSION: The use of the TLR2/6 agonist is a promising therapeutic approach in diseases with an imbalance in T cell responses, such as allergy and asthma. PMID- 20016196 TI - Regulation of CXCR/IL-8 in human airway epithelial cells. AB - BACKGROUND: Severe asthma is characterized by neutrophilic inflammation and high levels of interleukin (IL)-8. Airway epithelial cells play a pivotal role in the pathogenesis and chronicity of asthma. The objective of this work was to determine whether CXC receptors were involved in human small airway epithelial cell (SAEC) activity by incubating them with IL-8; the investigation also included a proteomic approach. METHODS: IL-6 and intercellular adhesion molecule 1 (ICAM-1) were assessed by ELISA and flow cytometry, respectively. CXCR-1 and CXCR-2 receptor mRNA and protein expressions were analyzed by RT-PCR, immunocytochemistry and flow cytometry. Cells were incubated with different concentrations (0-100 ng/ml) of IL-8. The involvement of both receptors was assessed using specific antibodies. RESULTS: Only the CXCR-1 receptor was expressed in SAECs. IL-8 (50 ng/ml, 12 h) induced the release of IL-6 and had no effect on ICAM-1. Supernatants analyzed by surface enhanced laser desorption ionization time of flight mass spectrometry (SELDI-TOF MS) showed very weak differences in peptide profiles. Interestingly, 4,820-m/z peptide release was detected in the presence of IL-8 and abolished by CXCR-1 antibody. DISCUSSION: The present study illustrated the fact that IL-8 mediated by CXCR-1 increased IL 6. We also highlight the usefulness of SELDI ProteinChip technology to confirm the potential variation of peptide profile. Moreover, we were able to detect the 4,820-m/z peptide secreted in vitro by human airway epithelial cells induced by IL-8 via CXCR-1 receptor. Determination of the protein secretion profile in response to inflammatory stimuli could be an important therapeutic strategy in severe asthma. PMID- 20016197 TI - Occupational endotoxin exposure reduces the risk of atopic sensitization but increases the risk of bronchial hyperresponsiveness. AB - BACKGROUND: Microbial exposures in both childhood and adult life are protective against atopy, allergic rhinitis and atopic asthma. In adults, this protective effect is paralleled by an increased prevalence of non-atopic asthma. This study was undertaken to investigate associations between occupational endotoxin exposure and atopic sensitization and bronchial hyperresponsiveness to methacholine (BHR) in agricultural workers. In addition, the role of atopy in endotoxin-related respiratory effects was studied. METHODS: Data were available for 427 farmers and agricultural industry workers, for whom airborne endotoxin exposure levels were estimated by 249 personal exposure measurements. Atopy was assessed as specific serum IgE to common inhalant allergens, and respiratory symptoms and personal characteristics by standardized questionnaires. BHR was determined in a subset of 113 subjects. Associations were adjusted for age, sex, smoking and living on a farm during childhood. RESULTS: Endotoxin exposure was positively associated with BHR and wheeze (p < 0.05). In contrast, endotoxin exposure was inversely associated with atopy and IgE to grass pollen (p < 0.001). The proportions of wheeze and BHR that were attributable to atopy were only 16.6 and 32.8%, respectively. CONCLUSIONS: High endotoxin exposure is a risk factor for BHR and wheeze, which were characterized by a predominantly non-atopic phenotype. At the same time, endotoxin exposure is related to a reduced risk of atopy and IgE to grass pollen in adults. It is unlikely that this is entirely a result of healthy worker selection, as significant inverse associations between endotoxin and IgE to grass pollen were found regardless of reported allergic symptoms. PMID- 20016198 TI - Exposure of intestinal epithelial cells to UV-killed Lactobacillus GG but not Bifidobacterium breve enhances the effector immune response in vitro. AB - BACKGROUND: Intestinal bacteria and intestinal epithelial cells (IEC) may modulate the mucosal immune response. In this study, immune modulation by Lactobacillus GG (LGG) and Bifidobacterium breve (Bb1, Bb2) in the presence or absence of IEC was addressed in an in vitro transwell co-culture model. METHODS: UV-killed LGG,Bb1, Bb2 or Toll-like receptor (TLR) 2 or nucleotide oligomerization domain (NOD) 2 ligands were added directly to unstimulated or anti-CD3/CD28-stimulated PBMC, or applied apically to human IEC (HT-29) co cultured with PBMC. A mixture of live bacteria was used as reference. The effect on T helper 1 (IFN-gamma, IL-12), T helper 2 (IL-13), inflammatory (TNF-alpha) and regulatory (IL-10) cytokine secretion was determined. RESULTS: Both UV-killed LGG and Bb enhanced IL-12, IFN-gamma, TNF-alpha and IL-10, and reduced IL-13 secretion when added directly to stimulated PBMC, similar to live bacteria. IEC reduced IL-13, IFN-gamma and IL-10 secretion by stimulated PBMC. Apically added LGG, TLR2 and NOD2 ligands,but not Bb, enhanced IFN-gamma, IL-12 and/or TNF-alpha secretion. Bacteria did not induce cytokine secretion when added to HT 29/unstimulated PBMC co-cultures, whereas direct incubations with PBMC did. CONCLUSION: UV-killed LGG as well as Bb supported a T helper 1 and/or regulatory phenotype when added directly to activated PBMC, similar to live bacteria. In contrast, LGG, TLR2 or NOD2 ligands - but not Bb - enhanced T helper 1 type cytokine secretion when added to IEC, while IL-10 secretion remained suppressed. Co-cultures combining IEC and PBMC may reveal differences between bacterial strains relevant for the in vivo situation. PMID- 20016199 TI - Human leukocyte antigen (DR1)-DQB1*0501 and (DR15)-DQB1*0602 haplotypes are associated with humoral responses to early food allergens in children. AB - BACKGROUND: Infants' immunological responses to cow's milk (CM) proteins, which in 2-3% result in allergy, may partially depend on genetic factors. We evaluated whether genes with immunological functions, i.e. human leukocyte antigen (HLA) II, the protein tyrosine phosphatase, non-receptor type 22 (PTPN22) and filaggrin, modulate immune responses to dietary antigens. METHODS: We analyzed 14 HLA class II haplotypes, the PTPN22 1858 SNP (R620W allele) and 5 known filaggrin null mutations from blood samples of 87 patients with CM allergy (CMA) and 76 control subjects (age 8.0-9.3 years). Serum levels of IgA, IgG, IgG1 and IgG4 antibodies to beta-lactoglobulin, alpha-casein and ovalbumin were measured with enzyme-linked immunosorbent assay, levels of IgE antibodies to CM, ovalbumin and birch with UniCap (Phadia, Uppsala, Sweden). RESULTS: In children with CMA, the HLA (DR15)-DQB1*0602 haplotype was associated with high levels of beta lactoglobulin-specific total IgG (p < 0.001) and IgG4 (p < 0.001) and alpha casein-specific total IgG (p = 0.003) and IgG4 (p = 0.002), but not among control subjects. (DR1/10)-DQB1*0501 was associated with lower levels of beta lactoglobulin-specific total IgG (p < 0.001) and IgG4 (p < 0.001), ovalbumin specific total IgG (p = 0.002) and IgG4 (p < 0.001), particularly in control subjects (p < 0.001). Six children with eczema (3 with CMA) had the filaggrin mutation del22824. PTPN22 was not associated with specific antibody responses or CMA. CONCLUSION: The HLA II, but not PTPN22 or filaggrin, genotype modulates humoral responses to early food allergens, whereas none of these genes was associated with CMA. PMID- 20016200 TI - Anaphylaxis to wheat flour-derived foodstuffs and the lipid transfer protein syndrome: a potential role of wheat lipid transfer protein Tri a 14. AB - BACKGROUND: Food allergy to wheat-derived foodstuffs is on the rise. Tri a 14, a wheat flour lipid transfer protein (LTP) allergen, has been described as a major allergen associated with baker's asthma and wheat food allergy. Cross-reactivity among LTP allergens leads to the so-called 'LTP syndrome'. METHODS: Eight adult patients showing anaphylaxis after ingestion of wheat-derived foodstuffs were selected. A homemade wheat extract, purified natural (n) and recombinant (r) Tri a 14, and peach fruit and Artemisia pollen LTP allergens Pru p 3 and Art v 3 were subjected to skin prick test, specific IgE determination (ELISA) and IgE immunodetection assays. RESULTS: All tests were positive in the 8 selected patients with the homemade extract. Positive skin prick test responses to nTri a 14, Pru p 3 and Art v 3 were found in 5/8, 6/8 and 4/4 patients, respectively. Specific IgE determined by ELISA assays was detected in 6 to nTri a 14 and rTri a 14, in 4 to Pru p 3 and in 3 to Art v 3 out of 8 individual sera tested, whereas all these sera showed IgE binding to nTri a 14 and Pru p 3 in immunodetection after SDS-PAGE separation. CONCLUSIONS: Tri a 14 seems to be a relevant allergen in patients with anaphylaxis after ingestion of wheat flour foodstuffs, according to in vitro and in vivo results. Clinical history of the analyzed patients, together with sensitization to peach Pru p 3 and Artemisia pollen Art v 3, suggests that 6 of them suffer from LTP syndrome. PMID- 20016201 TI - The diabetic antigen glutamic acid decarboxylase (GAD 65) in the human peripheral blood. AB - BACKGROUND: Glutamic acid decarboxylase (GAD 65) is a diabetes-associated antigen which is generally considered to be strictly intracellular. In order to better understand autoimmunity, this study demonstrates the appearance of GAD 65 in the peripheral human blood and presents implications for the diagnosis and therapy of some autoimmune diseases. METHODS: The GAD 65 molecules are detected by their interaction with monoclonal antibodies labeled with dyes in an experimental setup with fluorescence correlation spectroscopy (FCS). These interactions result in changes in Brownian motion measured as fluorescence fluctuations. Sera from 153 patients with diabetes mellitus type 1 and controls were investigated. To enable the representation of the molecule as a model for further discussions, we present structural visualizations of its hydrophobic properties, leading to possible interactions with the cell membrane lipids and epitope locations. RESULTS: The GAD65 antigen could be measured with a sensitivity of 2.65 microg/ml in 'clean systems' resulting from spiking experiments and human sera. The GAD 65 antigen could be identified in 8 patient sera: 4 children with diabetes mellitus type 1 and 4 adults initially taken as controls but who retrospectively showed signs of autoimmunity. CONCLUSION: We conclude that these findings are of significance for the concept of autoimmunity, i.e. in an initial step the immune system is primed by its accessibility to GAD 65. Our experimental results may also be important for the therapy of diabetes mellitus type 1 and other autoimmune diseases by the passive administration of GAD 65 antibodies. PMID- 20016202 TI - Site-specific connexin phosphorylation is associated with reduced heterocellular communication between smooth muscle and endothelium. AB - BACKGROUND/AIMS: Myoendothelial junctions (MEJs) represent a specialized signaling domain between vascular smooth muscle cells (VSMC) and endothelial cells (EC). The functional consequences of phosphorylation state of the connexins (Cx) at the MEJ have not been explored. METHODS/RESULTS: Application of adenosine 3',5'-cyclic monophosphate sodium (pCPT) to mouse cremasteric arterioles reduces the detection of connexin 43 (Cx43) phosphorylated at its carboxyl terminal serine 368 site (S368) at the MEJ in vivo. After single-cell microinjection of a VSMC in mouse cremaster arterioles, only in the presence of pCPT was dye transfer to EC observed. We used a vascular cell co-culture (VCCC) and applied the phorbol ester 12-O-tetradecanoylphorbol 13-acetate (PMA) or fibroblast growth factor-2 (FGF-2) to induce phosphorylation of Cx43 S368. This phosphorylation event was associated with a significant reduction in dye transfer and calcium communication. Using a novel method to monitor increases in intracellular calcium across the in vitro MEJ, we noted that PMA and FGF-2 both inhibited movement of inositol 1,4,5-triphosphate (IP(3)), but to a lesser extent Ca(2+). CONCLUSION: These data indicate that site-specific connexin phosphorylation at the MEJ can potentially regulate the movement of solutes between EC and VSMC in the vessel wall. PMID- 20016203 TI - Senescence of cultured porcine coronary arterial endothelial cells is associated with accelerated oxidative stress and activation of NFkB. AB - AIMS: Endothelial dysfunction occurs following multiple passaging in vitro,but the molecular mechanisms involved remain unidentified. The present study defined the genomic changes related to dysfunction in cultured senescent endothelial cells. METHODS AND RESULTS: Senescent cells were produced by multiple passaging of porcine coronary arterial endothelial cells for up to 4 weeks. Genomic and proteomic studies on cultured cells at the first passage (P1) and the fourth passage (P4) were performed. Senescence and decreased NO production were observed in cells and several signaling pathways - such as IFN/STAT, IGF, TGF-beta, cytoskeleton rearrangement and lipid metabolism - were altered at P4, as judged from the microarray analysis. The basal and stimulated (by TNF-alpha) levels of NFkappaB were augmented in senescent cells in electrophoretic mobility shift assays in association with increased oxidative stress, increased p53 protein stability, and activated apoptotic pathways. The increased oxidative stress was alleviated by treatment with the superoxide dismutase mimetic MnTMPyP. CONCLUSIONS: After multiple passaging in vitro, porcine coronary endothelial cells exhibited dysfunction and senescence associated with reduced proliferative capacity, increased oxidative stress, and activation of the NFkappaB and p53 signaling pathways. PMID- 20016204 TI - Effect of matrix metalloproteinase-9 knockout on vein graft remodelling in mice. AB - Long-term success in vein grafting for bypassing arteries blocked by atherosclerosis is limited by migration and proliferation of smooth muscle cells to form a neointima. Matrix metalloproteinases (MMPs), in particular MMP-2 and MMP-9, are implicated in neointimal formation by freeing smooth muscle cells from the cell-matrix contacts that normally restrict migration. We investigated the role of MMP-9 in vein grafts directly, using knockout mice. Vein grafts in MMP-9( /-) and wild-type mice had similar luminal and graft areas at 1, 4 and 8 weeks after engraftment, increasing with time. There was a relationship between the perimeter of the external elastic lamina and graft thickness (indicating graft remodelling) in MMP-9(-/-) mice at 1 week after surgery not apparent in control mice until later (r(2) = 0.933 for MMP-9(-/-) mice, r(2) = 0.040 for wild-type mice). Grafts in MMP-9(-/-) mice had 6-fold more pro- and active MMP-2 (p = 0.013, p = 0.026) than grafts in wild-type mice. Grafts from MMP-9(-/-) mice also had more collagen (p = 0.046 at 8 weeks), without any difference in cell number. Thus, while a lack of MMP-9 did not alter vein graft wall area or cellularity, grafts from MMP-9(-/-)mice accumulated more collagen and had earlier linear expansive remodelling, possibly due to an early compensatory increase in MMP-2. PMID- 20016206 TI - Identification of a 'snapshot' of co-expressed angiogenic markers in laser dissected vessels from unstable carotid plaques with targeted arrays. AB - BACKGROUND/AIMS: Angiogenesis is a feature of the atherogenic process, with intimal neovascularisation arising from vessels in the adventitia, adjacent to a plaque. Immature, leaky blood vessels from unstable plaques proliferate abnormally and, being poorly invested with smooth muscle cells, may contribute to instability of the plaque by facilitation of inflammatory cell infiltration and haemorrhagic complications. METHODS: We used laser-capture microdissection to isolate angiogenic areas of the extracellular matrix (containing CD105/flt-1 positive, fragile thin-walled vessels) and non-angiogenic vascular areas (CD105 negative, with smooth muscle cell covering) of complicated endarterectomy plaques, and specifically designed angiogenesis-TaqMan real-time PCR microarrays to identify gene expression. RESULTS: Important pro-angiogenic components, including Notch-3, delta-like-4 (DLL4), Tie-2, angiopoietin-1 (Angio-1) and receptor for advanced glycation end products (RAGE), and one anti-angiogenic factor, endostatin, were up-regulated in these regions. Immunohistochemistry demonstrated localisation within intimal, active (CD105-positive) microvessels and co-localisation of Notch-3 and DLL4/Tie-2 and Angio-1 in the same vessels indicating multiple/synergistic signalling mechanisms associated with vessel development. CONCLUSION: These data, although providing only a snapshot of information, demonstrate that plaque vascularisation occurs in the presence of multiple angiogenically active factors. Knowledge of their combined effects could help in the formulation of novel therapeutics designed to stabilise or prevent their formation in the treatment of atherosclerosis. PMID- 20016205 TI - Stainless steel ions stimulate increased thrombospondin-1-dependent TGF-beta activation by vascular smooth muscle cells: implications for in-stent restenosis. AB - BACKGROUND/AIMS: Despite advances in stent design, in-stent restenosis (ISR) remains a significant clinical problem. All implant metals exhibit corrosion, which results in release of metal ions. Stainless steel (SS), a metal alloy widely used in stents, releases ions to the vessel wall and induces reactive oxygen species, inflammation and fibroproliferative responses. The molecular mechanisms are unknown. TGF-beta is known to be involved in the fibroproliferative responses of vascular smooth muscle cells (VSMCs) in restenosis, and TGF-beta antagonists attenuate ISR. We hypothesized that SS ions induce the latent TGF-beta activator, thrombospondin-1 (TSP1), through altered oxidative signaling to stimulate increased TGF-beta activation and VSMC phenotype change. METHODS: VSMCs were treated with SS metal ion cocktails, and morphology, TSP1, extracellular matrix production, desmin and TGF-beta activity were assessed by immunoblotting. RESULTS: SS ions stimulate the synthetic phenotype, increased TGF-beta activity, TSP1, increased extracellular matrix and downregulation of desmin in VSMCs. Furthermore, SS ions increase hydrogen peroxide and decrease cGMP-dependent protein kinase (PKG) signaling, a known repressor of TSP1 transcription. Catalase blocks SS ion attenuation of PKG signaling and increased TSP1 expression. CONCLUSIONS: These data suggest that ions from stent alloy corrosion contribute to ISR through stimulation of TSP1-dependent TGF-beta activation. PMID- 20016207 TI - Autonomous effects of shear stress and cyclic circumferential stretch regarding endothelial dysfunction and oxidative stress: an ex vivo arterial model. AB - Cyclic circumferential stretch and shear stress caused by pulsatile blood flow work in concert, yet are very different stimuli capable of independently mediating endothelial function by modulating eNOS expression, oxidative stress (via production of superoxide anion) and NO bioavailability. Porcine carotid arteries were perfused using an ex vivo arterial support system for 72 h. Groups we created by combining normal (5%) and reduced (1%) stretch with high shear (6 +/- 3 dynes/cm(2)) and oscillatory shear (0.3 +/- 3 dynes/cm(2)) stress while maintaining a pulse pressure of 80 +/- 10 mm Hg. Oscillatory flow and reduced stretch both proved detrimental to endothelial function, whereas oscillatory flow alone dominated total endogenous vascular wall superoxide anion production. Yet, when superoxide anion production was analyzed in just the endothelial region, we observed that it was modulated more significantly by reduced cyclic stretch than by oscillatory shear, emphasizing an important distinction between shear- and stretch-mediated effects to the vascular wall. Western blotting analysis of eNOS and nitrotyrosine proved that they too are more significantly negatively modulated by oscillatory flow than by reduced stretch. These findings point out how shear and stretch stimulate regions of the vascular wall differently, affecting NO bioavailability and contributing to vascular disease. PMID- 20016208 TI - Porphyromonas gingivalis antigens and interleukin-6 stimulate the production of monocyte chemoattractant protein-1 via the upregulation of early growth response 1 transcription in human coronary artery endothelial cells. AB - BACKGROUND: Individuals with periodontitis have elevated serum levels of IL-6 and C-reactive protein and have been reported to have a significantly increased risk of developing cardiovascular disease. The transcription factor early growth response factor 1 (Egr-1) has been shown to play an important role in the development and progression of atherosclerosis. However, it is not known whether periodontal infection affects the expression of Egr-1 and subsequent endothelial cells expression of monocyte chemoattractant protein (MCP)-1, a key molecule of leukocyte chemoattraction into vessels. METHODS: Human coronary artery endothelial cells (HCAECs) were stimulated with either sonicated extracts from Porphyromonas gingivalis strains 381 or SU63, or a combination of IL-6 and soluble IL-6 receptor (IL-6/sIL-6R). The expression of Egr-1, and subsequently MCP-1, was then analyzed. The role of Egr-1 on MCP-1 expression was analyzed by siRNA transfection. RESULTS: Both P. gingivalis antigens and IL-6/sIL-6R stimulations upregulated the expression of Egr-1, with a more robust effect by IL 6/sIL-6R. Increased expression of Egr-1 coincided with MCP-1 production, and Egr 1 downregulation by siRNA suppressed this effect. CONCLUSION: These results clearly suggest that periodontal infection has the potential to affect HCAECs and hence contribute to the development of subsequent atherosclerosis. PMID- 20016209 TI - Thrombus versus wall biological activities in experimental aortic aneurysms. AB - BACKGROUND/AIM: The intraluminal thrombus (ILT) is considered to participate in abdominal aortic aneurysm (AAA) evolution. To assess whether this role proceeds via ILT influence on biological activity of the AAA wall, we studied the relationships between the levels of some relevant proteases and microparticles (MP) released by ILT versus wall in rat experimental AAAs. METHODS AND RESULTS: Two weeks after elastase perfusion, ILT and AAA wall were incubated in cell culture medium and studies were performed on conditioned media. As shown by gelatin zymography, ILT released higher amounts of MMP9 than the wall, whereas the level of MMP2 activation (active/pro) was similar. Levels of elastase and urokinase plasminogen activator, plasmin and MPs, determined, respectively, by casein zymography, substrate hydrolysis and flow cytometry, were higher in ILT than in wall. Aneurysm diameter positively correlated with wall MMP9 levels, MMP2 activation, plasmin activity and MP release. Moreover, wall and ILT levels of pro and active forms of MMP2, elastase and plasmin were positively correlated. Wall levels of MMP2 activation and plasmin activity also correlated with ILT weight. CONCLUSION: The present data suggest that, in this experimental model, ILT may contribute to AAA evolution via its influence on the level of aneurysmal wall protease activity. PMID- 20016210 TI - Role of gender in the associations of microalbuminuria with inflammatory markers in hypertensive subjects: a cross-sectional study. AB - BACKGROUND: Though the association between microalbuminuria (MA) and inflammatory markers has been studied, the possible gender differences in these associations have not yet been analyzed. Our study aims to analyze the role of gender in the associations of MA and inflammatory markers. METHODS: 1,060 hypertensive patients were assessed for MA (albumin-creatinine ratio), plasma levels of HsCRP (high sensitivity C-reactive protein), IL-18, and sCD40L (soluble CD40 ligand). Patients with diabetes mellitus, metabolic syndrome and overt nephropathy were excluded. RESULTS: Mean age was 46 +/- 9.6 years, with 560 males and 500 females. The prevalence of MA was 35.6% (n = 378). MA was associated with HsCRP (OR: 2.13, CI: 1.155-3.168, p = 0.001) and sCD40L (OR: 2.35, CI: 1.014-3.912, p = 0.013) in the premenopausal females, whereas in males (OR: 1.83, CI: 1.037-3.920, p = 0.023) and postmenopausal females (OR: 2.31, CI: 1.688-3.274, p = 0.031) MA was associated only with HsCRP and not with sCD40L or IL-18. CONCLUSIONS: Association between MA and HsCRP is consistent in all hypertensive patients. However, MA is associated with sCD40L only in premenopausal females and not in males and postmenopausal females. PMID- 20016211 TI - Angiotensin receptor blocker reduces proteinuria independently of blood pressure in children already treated with Angiotensin-converting enzyme inhibitors. AB - BACKGROUND/AIMS: Dual blockade of the renin-angiotensin system with angiotensin converting enzyme inhibitors (ACEI) and angiotensin receptor blockers (ARB) has higher antiproteinuric effects than single blockade in adults. In children, little is known on dual blockade of the renin-angiotensin system. The study investigates whether adding an ARB to proteinuric children already on ACEI reduces proteinuria. METHODS: A total of 10 children (median age 13.3 years) with chronic kidney disease and persistent proteinuria despite maximal dose of ACEI were included. Losartan was given at an initial dose 0.8 mg/kg/day. Proteinuria, blood pressure (BP) and renal function (glomerular filtration rate) were measured. RESULTS: Mean proteinuria decreased from 484 +/- 290 mg/mmol creatinine to 223 +/- 197 after 1-3 months of losartan treatment and remained stable at 234 +/- 153, 224 +/- 177 and 195 +/- 133 after 3-6, 6-12 months and at the last follow-up check (median 1.9 years, p < 0.05 for all visits vs. before treatment). The median percentage decrease in proteinuria was 66, 56, 44 and 66% during the study periods. No significant change in BP, glomerular filtration rate or serum potassium was observed. One child complained of rash, which led to discontinuation of losartan. CONCLUSION: Adding an ARB to current ACEI treatment can further reduce proteinuria in children with chronic kidney disease without affecting BP. PMID- 20016212 TI - Immunolocalization and mRNA expression of bone morphogenetic protein-6 in human clear cell renal carcinoma. AB - Bone morphogenetic proteins (BMPs) are members of the transforming growth factor beta superfamily of proteins. Dysregulation of BMP signaling has been suggested in the carcinogenesis of different organs. We determined BMP-6 mRNA and protein expression in localized human clear cell renal carcinoma (CCRC), obtained from 20 patients who underwent nephrectomy, by the real-time polymerase chain reaction and immunohistochemistry. 15/20 patients exhibited higher BMP-6 mRNA expression in malignant than in healthy renal tissue relative to the PBGD expression (p < 0.05). Immunostaining intensity for BMP-6 in healthy renal tissue ranged from 0 to 2 (average 0.9), as well as in renal clear cell carcinoma (average 1.1). Seven of 20 (35%) healthy tissue samples failed to stain with BMP-6 antibody, compared to 2/20 (10%) tumor samples (p < 0.05). BMP-6 immunostaining was positive in 18/20 CCRC samples. Staining was localized in the cytoplasm and/or membrane of malignant cells. Malignant tissue had significantly higher BMP-6 mRNA expression than healthy tissue. There was no significant correlation between BMP-6 mRNA and protein expression with disease presentation, disease progression and patients' characteristics. Long-term follow-up of our patients is needed to determine the possible role of increased expression of BMP-6 in CCRC. PMID- 20016213 TI - Peritoneal dialysis and hemodialysis in systemic lupus erythematosus patients: comparison of clinical outcomes. AB - BACKGROUND: This study compared peritoneal dialysis (PD) and hemodialysis (HD) outcomes between female systemic lupus erythematosus (SLE) patients with end stage renal disease (ESRD) due to lupus nephropathy. METHODS: 22 female SLE patients undergoing PD were compared with 14 female SLE patients receiving HD. Clinical outcomes and infective complications were reviewed. RESULTS: The overall mortality rate was much higher in the PD group (6/22) than in the HD group (1/14) (p = 0.027). PD patients had higher C-reactive protein level (37.1 +/- 41.4 vs. 6.7 +/- 9.5 mg/l, p = 0.037) and numbers of infectious episode (PD vs. HD: 1 episode per 33.16 patient-months vs. 1 episode per 118.26 patient-months, respectively, p = 0.046). Before the end of the observation period, HD patients had higher serum albumin (3.8 +/- 0.2 vs. 3.3 +/- 0.6 g/dl, p = 0.01). CONCLUSIONS: In female SLE patients with ESRD due to lupus nephropathy, clinical outcomes are better after undergoing HD than after undergoing PD. PMID- 20016214 TI - Telephone interview for cognitive status. PMID- 20016215 TI - Evaluation of the usability of a brief computerized cognitive screening test in older people for epidemiological studies. AB - BACKGROUND: Computerized cognitive testing has the potential to be an effective way to assess and monitor cognition in large neuroepidemiological studies. CogState is a game-like computerized test with demonstrated validity and reliability that has shown sensitivity to decline in older individuals over time. This study aimed to evaluate the serial usability of the test specifically within an older community cohort. METHODS: The test battery was administered to healthy volunteers aged 50 years and above at 3-month intervals over 12 months in a community setting. Test usability was examined in terms of acceptability, efficiency and stability. RESULTS: Of 301 subjects (age: 61.9 +/- 7.2 years), 87% completed the study. In addition, 85% completed the first test within the allowed time and passed integrity criteria with their performance improving and stabilizing at subsequent visits. The computerized battery required 15 min for administration on average, allowing 263 patients to be assessed on 5 occasions by 2 assessors. All tasks showed stability and a high test-retest reliability with serial administration. CONCLUSIONS: This computerized test was shown to have good acceptability, efficiency and stability for the repeated assessment of cognitive function in older people. Together with its demonstrated sensitivity to cognitive impairment and cognitive change, these data suggest that it would be a useful tool for application in neuroepidemiological studies. PMID- 20016216 TI - Incidence of Parkinson's disease in Bulgaria. AB - The incidence of Parkinson's disease (PD) was studied over a 3-year period (2002 2004) in the district of Plovdiv, Bulgaria with a mean population of 713,090. Of 663 possible cases, 244 patients diagnosed with PD were included in the study. The diagnosis PD was based on UK PD Society Brain Bank Criteria. The average crude incidence rate of PD was 11.44/ 100,000 person-years. After age adjustment to the general Bulgarian population, incidence was 11.65/100,000: 17.46 for men; 7.24 for women; 12.97 for the urban group and 9.55 for the rural group. Age specific incidence was 0.67/100,000 in the age range 40-44, reached a maximum of 112.73/100,000 for males and 52.58/100,000 for females in the age range 75-79 years and declined in the elderly. The present study estimates the incidence of PD in Bulgaria, which is similar to that in other European countries. The incidence rate in the Bulgarian population is twofold higher in men compared to women and slightly higher in the urban population compared with the rural population. PMID- 20016217 TI - Metabolic syndrome and cognition in patients with manifest atherosclerotic disease: the SMART study. AB - BACKGROUND: It is unclear whether the metabolic syndrome (MetS) increases risk of cognitive dysfunction beyond the level expected from its individual components. We examined the association of MetS with cognitive dysfunction and assessed whether MetS increased risk of cognitive dysfunction more than that of the sum of its individual components. METHODS: Data on 823 participants were used from the SMART-study, a cohort study among patients with atherosclerotic disease. MetS was defined according to the NCEP-ATPIII-criteria. Neuropsychological tests assessing memory, executive, and visuospatial functioning were performed. Regression analyses were performed to assess the association of MetS and its individual components with cognitive dysfunction. To examine whether MetS increased risk of cognitive dysfunction beyond its individual components we tested whether there was interaction on an additive scale by calculating the relative excess risk due to interaction (RERI). RESULTS: MetS was associated with increased risk of memory (OR 2.0, 95% CI 1.1-3.3) and visuospatial dysfunction (OR 2.3, 95% CI 1.4-2.7) but not with executive dysfunction. However, risk of memory and visuospatial dysfunction for having all MetS components was not greater than that of the sum of the individual components (RERI 0.2 and -0.9). CONCLUSIONS: In this population, MetS is related to increased risk of cognitive dysfunction but not more than that of the sum of its individual components. PMID- 20016218 TI - Mortality and predictors of death 1 month and 3 years after first-ever ischemic stroke: data from the first national acute stroke Israeli survey (NASIS 2004). AB - BACKGROUND: Despite declining age-adjusted stroke mortality rates, the disease remains the third most common cause of death in Israel. Based on a national survey, we examined mortality rates during the first 3 years after a first-ever acute ischemic stroke (IS) and the major predictors of short-term (1 month) and long-term (3 years) mortality. METHODS: In the National Acute Stroke Israeli Survey (NASIS 2004), data were collected on all hospitalized stroke patients in Israel during a 2-month period. Mortality rates for first-ever IS were assessed at 1 month and 3 years and predictors of death were evaluated using the Cox proportional hazard model. RESULTS: A total of 1,079 first-ever IS patients were included. Survival data were complete for over 99% of patients. Cumulative mortality rates were 9.9% at 1 month and 31.1% at 3 years. Of the survivors at 1 month, 23.5% did not survive for 3 years. At 1 month, the hazard ratio (HR) for death significantly increased with stroke severity. One-month mortality was also associated with a decreased level of consciousness (HR 2.9, 95% CI 1.7-5.1), total anterior circulation infarction (TACI); HR 4.9, 95% CI 1.6-15.2), temperature on admission (HR 1.5, 95% CI 1.1-2.1 per 1 degrees C), age (HR 1.04, 95% CI 1.02-1.07 per year) and glucose levels on admission (HR 1.003, 95% CI 1.001-1.006 per 1 mg/dl). Age-adjusted proportions of diabetes and chronic heart failure were considerably higher in the deceased compared with survivors at 3 years (48 vs. 38 and 21 vs. 9%, respectively). In the multivariate survival analyses, predictors of death at 1 month also predicted death at 3 years; however, history of dementia (HR 1.5, 95% CI 1.0-2.4), diabetes (HR 1.6, 95% CI 1.0-2.4), peripheral artery disease (HR 1.7, 95% CI 1.1-2.8), chronic heart failure (HR 1.6, 95% CI 1.1-2.4) and malignancy (HR 1.7, 95% CI 1.1-2.7) were additional predictors of long-term mortality for patients surviving the first month after stroke. CONCLUSIONS: Approximately one third of patients did not survive 3 years after the first-ever IS. While age and markers of severe stroke were the major predictors of death at 1 month, comorbidities and variables associated with atherosclerotic vascular disease predicted long-term mortality. Improved control of these factors can potentially reduce long-term mortality in stroke victims. PMID- 20016219 TI - Prediction and prevention of death after first-ever ischemic stroke. PMID- 20016220 TI - Hypertension increases the probability of Alzheimer's disease and of mild cognitive impairment in an Arab community in northern Israel. AB - BACKGROUND: We aimed to determine whether vascular risk factors are associated with Alzheimer's disease (AD) and mild cognitive impairment (MCI) in an elderly Arab population. METHODS: An Arabic-speaking team performed a door-to-door survey of consecutive residents aged > or =65 years. We estimated the odds of AD or MCI versus normal controls as a function of age, gender, education and presence of vascular factors by multinomial logistic regression with interactions. RESULTS: Out of 767 subjects (54% men), 444 were cognitively normal, 234 had MCI and 89 had AD. AD was significantly associated with hypertension (p = 0.01; OR = 2.08; 95% CI: 1.18-3.65), age (p < 0.0001; OR = 1.19; 95% CI: 1.14-1.24), female gender (p = 0.0016; OR = 3.06; 95% CI: 1.53-6.15) and education (p = 0.0002; OR = 0.75; 95% CI: 0.65-0.88). MCI was significantly associated with hypertension (p = 0.0042; OR = 1.69; 95% CI: 1.25-2.44), age (p < 0.0001; OR = 1.06; 95% CI: 1.03 1.09) and education (p < 0.0001; OR = 0.76; 95% CI: 0.71-0.83), but not with gender. CONCLUSIONS: Hypertension, older age and low education significantly increase the probability of AD and MCI. The effect of hypertension on the odds of AD versus controls is over and above the effects of age, gender and education. For MCI versus controls there is no gender effect, and the effect of hypertension is over and above the effects of age and education. PMID- 20016221 TI - Cdc42 regulates myocardin-related transcription factor nuclear shuttling and alpha-smooth muscle actin promoter activity during renal tubular epithelial mesenchymal transition. AB - BACKGROUND/AIMS: Epithelial-mesenchymal transition of tubular cells into alpha smooth muscle actin (SMA)-expressing myofibroblasts is a central mechanism in tubulointerstitial fibrosis. Previously, a 'two-hit' model was proposed for epithelial-mesenchymal transition wherein an initial injury of the intercellular contacts and TGF-beta(1) are both required for SMA protein expression in LLC-PK1 cells. The Rho-Rho kinase-myosin light chain-myocardin-related transcription factor (MRTF)-serum response factor (SRF) pathway and Rac1, p21-activated kinase (PAK) and p38 were described as important regulators of MRTF localization and SMA expression. Cdc42 is another small G protein situated upstream of PAK and p38, and is activated upon cell contact disassembly. Here, we investigated its potential role in the regulation of MRTF nuclear shuttling and in the regulation of the SMA promoter. RESULTS: Transfection of a constitutive active (CA) Cdc42 construct alone induced the activation of the SMA promoter. The dominant negative (DN) Cdc42 construct prevented the activation of the promoter induced by cell contact disassembly, and reduced the combined effect of cell contact disruption and TGF-beta(1). SRF showed a marked nuclear accumulation in CA Cdc42-transfected cells. Cdc42 induced the nuclear translocation of MRTF, while DN Cdc42 inhibited its nuclear translocation induced by cell contact disassembly. Blocking PAK, MRTF and p38 by the corresponding DN constructs blunted the effects of CA Cdc42 on the SMA promoter. CONCLUSION: Cdc42 is involved in the regulation of SMA promoter activation through PAK, p38, MRTF and SRF. Cdc42 may be an important regulator of MRTF cellular localization. PMID- 20016222 TI - Response of chicken ductus arteriosus to hypercarbic and normocarbic acidosis. AB - BACKGROUND: Changes in pH can have profound effects on vascular tone and reactivity, but their influence on the ductus arteriosus (DA) remains unknown. OBJECTIVE: To analyzethe effects of hypercarbic and normocarbic acidosis in the reactivity of the chicken DA. METHODS: DA rings from 19-day chicken fetuses (total incubation time, 21 days) were mounted in a wire myograph for isometric tension recording. RESULTS: In DA rings (pulmonary side) stimulated with O(2), norepinephrine (NE), KCl, or U46619, changes from control conditions (5% CO(2), 24 mM NaHCO(3), pH 7.4) to 7.5% CO(2) (pH 7.25) or 10% CO(2) (pH 7.14) induced a concentration-dependent relaxation that reached 43.0% (SD 21.3) of the O(2)-, 28.6% (SD 23.1) of the NE-, 10.4% (SD 18.7) of the KCl-, and 6.8% (SD 12.6) of the U46619-induced contraction. Hypercarbic-acidosis-induced relaxation was impaired by the non-selective K(+) channel blocker tetraethylammonium or the BK(Ca) channel inhibitor iberiotoxin. Normocarbic acidosis (5% CO(2), 12 mM NaHCO(3), pH 7.13) induced transient relaxation of the DA, which was not affected by the presence of tetraethylammonium or iberiotoxin. Euhydric hypercarbia (10% CO(2), 48 mM NaHCO(3), pH 7.46) induced a transient contraction of the DA. CONCLUSIONS: Our results indicate that the chicken DA is very sensitive to changes in extracellular pH, and that stimulation of BK(Ca) channels may account for the ductal-relaxing effects of hypercarbic acidosis. PMID- 20016223 TI - Genetic association analysis of functional polymorphisms in neuronal nitric oxide synthase 1 gene (NOS1) and mood disorders and fluvoxamine response in major depressive disorder in the Japanese population. AB - BACKGROUND/AIM: Nitric oxide has been reported to play a role in neural transmitter release and N-methyl-D-aspartate receptor activation, as well as to be related to oxidative stress. Abnormalities in both of these mechanisms are thought to be involved in the pathophysiology of mood disorders including major depressive disorder (MDD) and bipolar disorder (BP). In addition, several lines of evidence support an association between abnormalities in neuronal nitric oxide synthases (nNOS) and mood disorders. Therefore, we studied the association of nNOS gene (NOS1) and mood disorders and the efficacy of fluvoxamine treatment in Japanese MDD patients. MATERIALS AND METHODS: Using a single nucleotide polymorphism (SNP; rs41279104, also called ex1c), we conducted a genetic association analysis of case-control samples (325 MDD patients, 154 BP patients and 807 controls) in the Japanese population. In addition, we performed an association analysis between NOS1 and the efficacy of fluvoxamine treatment in 117 MDD patients. We defined a clinical response as a decrease of more than 50% in baseline SIGH-D (Structured Interview Guide for the Hamilton Rating Scale for Depression) score within 8 weeks, and clinical remission as an SIGH-D score of less than 7 at 8 weeks. RESULTS: We did not detect a significant association between NOS1 and MDD, BP or the fluvoxamine therapeutic response in MDD in allele/genotype-wise analysis. CONCLUSIONS: We did not detect an association between only one marker (rs41279104) in NOS1 and Japanese mood disorder patients and fluvoxamine response, but sample sizes were probably too small to allow a meaningful test. Moreover, because we did not perform an association analysis based on linkage disequilibrium and a mutation scan of NOS1, a replication of the study using a larger sample and based on linkage disequilibrium may be required for conclusive results. PMID- 20016224 TI - Dopaminergic pathway gene polymorphisms and genetic susceptibility to schizophrenia among north Indians. AB - OBJECTIVE: Understanding the etiology and pathogenesis of schizophrenia has been difficult due to the complex inheritance patterns, genetic heterogeneity and varied multiple nonlinear interactions between genes. Several lines of evidence indicate the involvement of neurotransmitter dopamine in the pathophysiology of this disorder. To analyze such a possible role of dopaminergic pathway gene polymorphisms, we used a case-control approach. METHOD: We genotyped a total of 31 potential single nucleotide polymorphism/variable number of tandem repeat markers from 9 candidate genes including the dopamine receptors and metabolizing enzymes (synthesis and degradation) in 215 schizophrenia cases and 215 healthy controls from North India. RESULTS: A nominally significant allelic association was observed in case of the catechol-O-methyltransferase rs362204 -/G (p = 0.028) marker whereas nominally significant genotypic associations were seen for tyrosine hydroxylase rs6356 A/G (p = 0.04) and dopamine beta-hydroxylase rs1108580 A/G (p = 0.025) following the case-control approach. Several significant haplotypic associations were observed from dopamine beta-hydroxylase, catechol-O-methyltransferase, and dopamine receptor D(2) genes. A significant interaction of tyrosine hydroxylase rs6356 A/G and catechol-O-methyltransferase rs362204 -/G markers was also observed following binary logistic regression analysis (p = 0.009). CONCLUSIONS: A contribution of dopaminergic pathway gene polymorphisms to schizophrenia seems possible in our sample set. However, considering the marginal levels of associations, interpopulation comparisons and replicate studies are warranted. PMID- 20016225 TI - Association of brain-derived neurotrophic factor genetic Val66Met polymorphism with severity of depression, efficacy of fluoxetine and its side effects in Chinese major depressive patients. AB - BACKGROUND: Preclinical studies have shown that brain-derived neurotrophic factor (BDNF) may be involved in antidepressant action, and the BDNF gene has been suggested to be involved in the pharmacological treatment of major depressive disorder (MDD). In this study, the relationship between BDNF Val66Met polymorphism (Single Nucleotide Polymorphism Database ID: rs6265) and severity of depression, efficacy of fluoxetine and its side effects was tested in Chinese patients with MDD. METHODS: Patients with MDD took the oral selective serotonin reuptake inhibitor (SSRI) fluoxetine (20 mg/day) for 6 weeks. Its clinical efficacy and side effects were measured by the 17-item Hamilton Rating Scale for Depression and the Treatment-Emergent Symptoms Scale (TESS), respectively. The patients were genotyped for Val66Met polymorphism of the BDNF gene. RESULTS: In the multivariate regression analysis, there was no significant association between severity of depression and BDNF Val66Met polymorphism. There was no association between efficacy of fluoxetine and BDNF Val66Met polymorphism, but there was a marginal positive suggestion that heterozygous patients tended to have a better remission with fluoxetine in comparison with homozygous analogs. Insomnia and decreased sexual desire, side effects of fluoxetine, may have an association with the BDNF Val66Met polymorphism, and Met allele carriers showed a lower incidence of these side effects. CONCLUSIONS: These results indicate that there was a lack of association between severity of depression and BDNF Val66Met polymorphism in Chinese patients with MDD. The BDNF Val66Met polymorphism may play a major role in the efficacy and side effects of SSRI (fluoxetine) in Chinese patients with MDD. PMID- 20016226 TI - Sustained attention and planning deficits but intact attentional set-shifting in neuroleptic-naive first-episode schizophrenia patients. AB - INTRODUCTION: The nature of deficits in tests of sustained attention, planning and attentional set-shifting has not been investigated in neuroleptic-naive first episode (FE) schizophrenia patients. Based on previous literature of chronic and medicated FE schizophrenia patients, we predicted that the neuroleptic-naive patients would show deficits in these cognitive processes. METHODS: Twenty-nine neuroleptic-naive FE schizophrenia patients and 33 healthy controls - matched by age, gender, and nicotine consumption - performed 3 tests from the Cambridge Automated Neuropsychological Test Battery (CANTAB) thought to measure these cognitive processes: the Rapid Visual Information Processing task (RVIP, sustained attention), the Stockings of Cambridge task (SOC, planning), and the Intradimensional/Extradimensional set-shifting task (IDED, attention shifting). RESULTS: The patients were significantly impaired in the sensitivity index (A') of the RVIP, and in the number of problems solved with minimum moves on the SOC. Nevertheless, the groups did not differ regarding the number of participants who failed at the crucial extradimensional shift stage of the IDED. CONCLUSION: Sustained attention and planning abilities are already impaired in neuroleptic naive FE schizophrenia patients, whereas set-shifting abilities as measured with the IDED task seem to be intact at illness onset. Since chronic schizophrenia patients have been shown to have impaired IDED performance, we tentatively propose that IDED performance deteriorates over time with illness chronicity and/or medication. PMID- 20016227 TI - Oxaliplatin-induced lhermitte's phenomenon as a manifestation of severe generalized neurotoxicity. AB - OBJECTIVES: Lhermitte's phenomenon, characterized by 'electric-shock' sensations precipitated by neck flexion, may develop during oxaliplatin treatment. Limited cases have been described previously and the pathophysiology underlying Lhermitte's phenomenon in oxaliplatin-treated patients has not been established. METHODS: Patients who developed Lhermitte's phenomenon during oxaliplatin therapy were investigated by neurological examination, neurotoxicity grading and conventional nerve conduction studies (NCS). Structural (magnetic resonance imaging) and functional (somatosensory evoked potentials) spinal assessment was also undertaken. Sensory nerve excitability recordings were performed longitudinally across treatment to investigate ion channel function. RESULTS: Five oxaliplatin-treated patients reported Lhermitte's phenomenon, with a mean cumulative dose of 861 +/- 84 mg/m(2) oxaliplatin (range 574-1,100 mg/m(2)). NCS revealed severe sensory neuropathy in all patients. There was no evidence of structural or functional spinal cord damage. Nerve excitability studies revealed progressive alterations in sensory excitability throughout treatment, consistent with oxaliplatin-induced nerve dysfunction. In patients with Lhermitte's phenomenon, refractoriness was reduced to -14.4% (confidence interval, CI: -20.5 to -8.4%) by late treatment, a significantly greater reduction than in oxaliplatin-treated patients who did not develop Lhermitte's phenomenon (-2.7%; CI: -7.6 to 2.2; p = 0.013). CONCLUSIONS: Lhermitte's phenomenon represents a severe presentation of oxaliplatin-induced neurotoxicity, associated with generalized nerve dysfunction in the absence of structural spinal abnormalities. PMID- 20016228 TI - A phase II feasibility study of weekly paclitaxel in heavily pretreated advanced gastric cancer patients with poor performance status. AB - PURPOSE: We investigated the efficacy and safety of weekly paclitaxel monotherapy in previously treated patients with advanced gastric cancer (AGC) and poor performance status (PS) according to the Eastern Cooperative Oncology Group (ECOG). PATIENTS AND METHODS: Patients with evaluable disease who failed at least one previous chemotherapy and had a PS of 2-3, received paclitaxel 70 mg/m(2) on days 1, 8 and 15 every 4 weeks. RESULTS: The median overall survival (OS) was 5.5 months (95% confidence interval, CI, 3.3-7.8) and progression-free survival was 2.1 months (95% CI, 1.2-3.0). The overall response rate was 3.8% and the disease control rate was 25.0%. Treatment-related toxicities were tolerable. OS was 7.1 (95% CI, 5.4-9.5) and 3.7 months (95% CI, 2.1-5.3) for patients with PS-ECOG 2 and 3, respectively (p < 0.001). When evaluated according to the previous treatment, OS was 5.1 (95% CI, 3.3-7.0) and 6.5 months (95% CI, 3.8-9.3) for patients receiving two and three or more lines of treatment, respectively (p = 0.815). With multivariate analysis, PS was a significant factor for OS. CONCLUSION: Survival in patients treated with weekly paclitaxel monotherapy was comparable to other second- or third-line chemotherapies for AGC, with acceptable toxicities in previously treated patients with poor PS. PMID- 20016229 TI - Predicting therapy outcome with quantitative PET: what is needed and what can be done? PMID- 20016230 TI - Differential therapy for diffuse large B-cell lymphoma with different cells of origin. PMID- 20016231 TI - Malignant melanoma and pregnancy. PMID- 20016232 TI - The effect of biweekly CHOP and standard CHOP in different subgroups of diffuse large B-cell lymphoma. AB - BACKGROUND: Diffuse large B-cell lymphoma (DLBCL) is the most common type of non Hodgkin's lymphoma. Conventional chemotherapy (CHOP: cyclophosphamide, doxorubicin, vincristine, prednisone) seems unsatisfactory, so modifications of CHOP are used to improve the efficacy. DLBCL is a highly variable tumor, with different responses to therapy. The aim of this study is to explore the association between immunophenotype and treatment response. PATIENTS AND METHODS: We analyzed the expression of Bcl-6, CD10, and MUM1 in 130 cases of DLBCL using immunohistochemistry. The cases were subdivided into germinal center B-cell-like (GCB) and non-GCB subtypes, and were randomly assigned to receive either 6-8 cycles of CHOP every 2 weeks or standard CHOP every 3 weeks. RESULTS: After a median followup duration of 40 months, 3-year overall survival of biweekly CHOP was better than standard CHOP in the non-GCB group (58.3 vs. 38.6%, p < 0.05). However, the therapeutic effect of biweekly CHOP and standard CHOP was not different in the GCB group (64.8 vs. 57.5%, p > 0.05). CONCLUSIONS: Immunohistochemistry analysis of different subgroups is useful to find the suitable therapy. Biweekly CHOP showed higher efficacy than standard treatment in the non-GCB subgroup. PMID- 20016233 TI - Limited predictive value of FDG-PET for response assessment in the preoperative treatment of esophageal cancer: results of a prospective multi-center trial (SAKK 75/02). AB - BACKGROUND: Only responding patients benefit from preoperative therapy for locally advanced esophageal carcinoma. Early detection of non-responders may avoid futile treatment and delayed surgery. PATIENTS AND METHODS: In a multi center phase ll trial, patients with resectable, locally advanced esophageal carcinoma were treated with 2 cycles of induction chemotherapy followed by chemoradiotherapy (CRT) and surgery. Positron emission tomography with 2[fluorine 18]fluoro-2-deoxy-d-glucose (FDG-PET) was performed at baseline and after induction chemotherapy. The metabolic response was correlated with tumor regression grade (TRG). A decrease in FDG tumor uptake of less than 40% was prospectively hypothesized as a predictor for histopathological non-response (TRG > 2) after CRT. RESULTS: 45 patients were included. The median decrease in FDG tumor uptake after chemotherapy correlated well with TRG after completion of CRT (p = 0.021). For an individual patient, less than 40% decrease in FDG tumor uptake after induction chemotherapy predicted histopathological non-response after completion of CRT, with a sensitivity of 68% and a specificity of 52% (positive predictive value 58%, negative predictive value 63%). CONCLUSIONS: Metabolic response correlated with histopathology after preoperative therapy. However, FDG-PET did not predict non-response after induction chemotherapy with sufficient clinical accuracy to justify withdrawal of subsequent CRT and selection of patients to proceed directly to surgery. PMID- 20016234 TI - Detection of lymphovascular invasion in vulvar cancer by D2-40 (podoplanin) as a predictor for inguinal lymph node metastases. AB - BACKGROUND: Lymphatic vessel invasion (LVI) plays a major role in the spread of vulvar cancer and predicts regional lymph node metastasis. D2-40, a monoclonal immunohistochemical marker might be able to increase the detection rate of LVI compared to conventional hematoxylin-eosin (HE) staining. The aim of the study was to evaluate the suitability of D2-40 for the prediction of regional lymph node metastases. PATIENTS AND METHODS: Immunohistochemical staining with D2-40 was performed on formalin-fixed, paraffin-embedded tissue sections of 32 patients with squamous cell carcinoma of the vulva. Slides were screened for the presence of LVI. Correlation with clinico-pathological features including LVI as retrieved by routine HE-stained sections was assessed. RESULTS: Immunostaining with D2-40 significantly (p = 0.019) increased the frequency of detection of lymphatic invasion compared to conventional HE staining. LVI was correctly identified by D2 40 (D2-40+ LVI) in 65.6% of tumor specimens as compared to 40.6% by routine HE staining (HE+ LVI). D2-40+ LVI significantly (p = 0.026) predicted inguinal lymph node metastases. CONCLUSIONS: Immunostaining with D2-40 significantly increased the frequency of detection of lymphatic invasion compared to conventional HE staining in squamous cell carcinomas of the vulva. D2-40+ LVI is a strong predictor for inguinal lymph node metastases. PMID- 20016235 TI - Single-center management of 136 patients with cancer of unknown primary site (CUP syndrome) over a period of 10 years. AB - BACKGROUND: Cancer of unknown primary site (CUP syndrome) is a particular challenge in oncology which occurs in about 5-10% of cancer patients. Here, we investigated clinicopathological and prognostic factors in patients with CUP syndrome in a retrospective analysis. PATIENTS AND METHODS: 136 patients with CUP syndrome who were treated in our Department of Oncology and Hematology were analyzed over a period of 10 years. Clinical and histopathological characteristics, response to chemotherapy, survival and prognostic factors were investigated in a retrospective analysis. RESULTS: 83 of the patients (61%) received first-line chemotherapy, which induced an overall response rate of 19%. Altogether 37 different chemotherapy regimens were used. Median overall survival of all patients was 7.9 months. In multivariate Cox regression analysis, gender, Karnofsky performance status, treatment modality and extent of disease were identified as independent prognostic factors. CONCLUSIONS: Our analysis showed a poor prognosis for patients with CUP syndrome. The response rate to chemotherapy was low with no significant benefit for any of the investigated cytotoxic agents. Newer diagnostic and therapeutical approaches might contribute to an improvement of prognosis, and their value is currently investigated in prospective studies. PMID- 20016236 TI - Advanced metastatic melanoma during pregnancy: a multidisciplinary challenge. AB - BACKGROUND: Metastatic melanoma during pregnancy represents a life-threatening situation not only for the mother but also for the fetus due to aggressive therapy and potential maternal-fetal metastasis. CASE REPORT: We report the case of a 37-year-old woman with advanced metastatic malignant melanoma during her first pregnancy, with a review of the literature. In this case, a tight and primarily interdisciplinary obstetrical and dermatological case management enabled the delivery of a small but healthy premature infant in the 29th week of gestation by planned Cesarean section. However, due to progressive disease, the mother died only 10 weeks after the delivery of the baby. CONCLUSION: Sufficient perinatal and oncologic experience provided, diagnostic and surgical interventions as well as radiotherapy and chemotherapy in metastatic melanoma disease are feasible and relatively safe even during pregnancy. PMID- 20016237 TI - Ovarian metastasis of a lymphoma presenting as primary ovarian cancer. AB - BACKGROUND: Ovarian metastases are being found in a variety of different malignant diseases. CASE REPORT: A clinical case of a woman with a pelvic tumor and elevated CA125 levels, and suspected primary ovarian carcinoma is presented. Surgery revealed a metastasis of a B-cell non-Hodgkin's lymphoma. CONCLUSIONS: Literature research shows that elevated CA125 levels are often found in lymphomas. PMID- 20016238 TI - Cisplatin-associated occlusion of the internal carotid artery. AB - BACKGROUND: Cisplatin is a highly effective antineoplastic agent used also in germ cell cancer. Thromboembolic complications like pulmonary embolism, myocardial infarction and stroke have occasionally been reported. CASE REPORT: A 46-year-old man with seminoma and without any cerebrovascular risk factors developed an acute stroke with complete right-sided hemiparesis and global aphasia, during the second cycle of cisplatin-based chemotherapy. After exclusion of an intracerebral bleeding, a systemic thrombolysis was performed. Further diagnostics revealed an acute occlusion of the left carotid internal artery with infarction in the territory of the middle cerebral artery. The patient slowly recovered; however, a moderate aphasia and partial hemiparesis persisted whereas the tumor was in complete remission. Months later, the patient developed symptomatic epilepsy. CONCLUSIONS: Ischemic stroke associated with cisplatin based chemotherapy is rare, but may be disabling or even fatal. With regard to the literature, several precautions are discussed to minimize the risk of these major side effects. PMID- 20016239 TI - Prostate cancer with metastasis to the omentum and massive ascites: a rare manifestation of a common disease. AB - BACKGROUND: The most common sites of metastasis in prostate cancer include bone and regional lymph nodes followed by lung, liver, and brain. Peritoneal metastasis without skeletal involvement is extremely rare. CASE REPORT: We present herein a patient with hormone refractory prostate cancer with peritoneal metastasis accompanied by ascites but without bone metastasis. The patient initially experienced an excellent response to docetaxel-based chemotherapy. CONCLUSIONS: Prostate cancer can present with distant metastasis in unexpected sites. The lack of skeletal involvement does not exclude the possibility of distant metastases. The presence of ascites may indicate peritoneal disease which could be responsive to current standard chemotherapy. PMID- 20016240 TI - Preoperative detection of cervical lymph node metastases in papillary thyroid cancer: a surgical perspective. AB - Despite its relatively benign biological behavior, papillary thyroid cancer is frequently associated with cervical lymph node metastases at the time of diagnosis. These metastases have a limited impact on overall survival, but are recognized as a significant risk factor for locoregional recurrence of the disease. This may significantly alter quality of life, and may require further therapeutic interventions which may be associated with increased morbidity. Therefore, preoperative identification of cervical lymph node metastases is of particular importance and allows optimal and effective treatment at the time of initial surgery. Clinical examination remains important but lacks sensitivity. Neck ultrasonography is currently the most useful method to detect cervical lymphadenopathy. Fine-needle aspiration (for cytology and thyroglobulin measurement), usually under ultrasonographic guidance, may confirm the diagnosis of lymph node metastases. Other imaging methods (including computed tomography, magnetic resonance imaging, positron emission tomography) should be used selectively. A compartment-oriented cervical lymph node dissection should be performed at the time of thyroidectomy if preoperative evaluation reveals cervical lymphadenopathy. PMID- 20016242 TI - Three-dimensional computed tomography imaging of the eustachian tube lumen in patients with patulous eustachian tube. AB - BACKGROUND: The eustachian tube (ET) has a complicated anatomy, which is related to middle ear pathology. The anatomy of the ET has been well investigated in cadavers, but may not accurately reflect that in living subjects. Three dimensional (3D) computed tomography (CT) was used to examine the ET in patients with patulous ET. METHOD: The subjects were 35 patients (17 males, aged 59 +/- 16 years, and 18 females, aged 44 +/- 15 years) with patulous ET which could be continuously traced from the pharyngeal orifice to the tympanic orifice (53 ears). CT was performed in the sitting position at rest, followed by 3D reconstruction. The course of the ET from the middle ear to the pharynx was visualized in 3D surface images. Morphological measurements of the curvature angle (angle C) and the inclination angle (angle I) were determined from 3D coordinates on the CT images. RESULTS: The full length of the ET could be visualized in most patients suffering from patulous ET in the sitting position. Angle C was 21.7 +/- 5.8 degrees in the patients aged <40 years (n = 20), which is significantly larger than 16.7 +/- 6.8 degrees in the patients aged >59 years (n = 18; Student's t test, p < 0.05). Angle I was 26.5 +/- 4.2 degrees in the patients aged <40 years (n = 20), also significantly larger than 21.6 +/- 5.8 degrees in the patients aged >59 years (n = 18; Student's t test, p < 0.05). CONCLUSION: CT in the sitting position is a potential diagnostic tool of the ET and middle ear pathology. PMID- 20016243 TI - Lipid biology. AB - The field of 'lipidology' has been building momentum in recent years and continues to evolve with the field of medicine as a whole. The field has been growing rapidly in response to unfortunate demographic trends that place an increasing number of people, in particular in the United States, at risk for coronary heart disease and epidemic levels of obesity, which coincide with a surge in new cases of diabetes mellitus. In addition, one of the major causes of pancreatitis in developed countries includes hyperlipidemia. Therefore, lipid biology is important in the context of the pancreas, both during our normal digestive process and its impact on diseases affecting the pancreas, such as diabetes, pancreatitis, and cancer. PMID- 20016244 TI - Primers on molecular pathways - lipoxygenases: their role as an oncogenic pathway in pancreatic cancer. AB - Different evidence supports a functional role of enzymes involved in lipid metabolic pathways, such as lipoxygenases (LOXs) and their metabolite derivatives, in carcinogenesis. LOX enzymes catalyze the dioxygenation of arachidonic acid into hydroxyperoxyeicosatetraenoic acids, which is followed by their conversion to their corresponding eicosanoids as hydroxyeicosatetraenoic acids, leukotrienes, lipoxins and hepoxilins, which in turn act as cellular messengers. Subcellular LOX enzyme localization varies according to the LOX and cellular type regulating different cell functions. LOX enzymes or their products may exert their biological effects in different modes, either intracellular or in other cells. Numerous clinical studies on expression of LOXs in human tumors as well as in animal models indicate different roles of distinct LOX isoforms in carcinogenesis. In fact, different LOXs exhibit either protumorigenic or antitumorigenic activities and modulate the tumor response in a tissue-specific manner. Moreover, the LOX pathways are involved in the spread and metastasis of several cancers, including pancreas, through the activation of several cellular signaling pathways which modify gene expression affecting cellular proliferation, survival, migration and extracellular matrix production. In this review we focus on the important role and different mechanisms of action of LOX pathways in the regulation of pancreatic cancer initiation and progression. A novel approach for pancreatic cancer chemoprevention would involve targeting LOX activities, alone or in combination with other pathways as a major anticancer strategy. PMID- 20016245 TI - Puerarin inhibits adhesion molecule expression in tnf-alpha-stimulated human endothelial cells via modulation of the nuclear factor kappaB pathway. AB - BACKGROUND: The isoflavone puerarin is the most abundant isoflavone-C-glucoside extracted from the root (radix puerariae) of the plant Pueraria lobata and possesses many biological activities. In this report, the ability of puerarin to modulate intercellular adhesion molecule 1 (ICAM-1), vascular cell adhesion molecule 1 (VCAM-1) and endothelial leukocyte adhesion molecule 1 (E-selectin), and to induce changes in the nuclear factor kappaB (NF-kappaB) pathway in human umbilical vein endothelial cells (HUVECs) was examined. METHODS: The protein and mRNA levels of tumor-necrosis-factor-alpha (TNF-alpha)-induced ICAM-1, VCAM-1 and E-selectin were determined in HUVECs. Inhibitor kappaB (I-kappaB) phosphorylation and p65 NF-kappaB expression in HUVECs were also examined. RESULTS: Puerarin inhibited the expression of TNF-alpha-induced ICAM-1, VCAM-1 and E-selectin proteins and mRNAs in HUVECs. Subsequently, we determined that the inhibition of ICAM-1, VCAM-1 and E-selectin expression was due to a dose-dependent suppression of phosphorylation and degradation of I-kappaB, which resulted in a reduction of p65 NF-kappaB nuclear translocation. CONCLUSION: These data suggested that the effect of puerarin-mediated inhibition of TNF-alpha-induced ICAM-1, VCAM-1 and E selectin expression is attributed to suppressed NF-kappaB activation on the transcriptional level. PMID- 20016246 TI - Evidence for the involvement of glutamatergic and neurokinin 1 receptors in the antinociception elicited by tramadol in mice. AB - This study was designed to evaluate the role of spinal glutamatergic receptors and neurokinin 1-mediated pathway in the antinociception elicited by tramadol in mice. Tramadol (1-50 mg/kg), given intraperitoneally (i.p.), produced significant inhibition of the biting behavior induced by intrathecal injection (i.t.) of glutamate (175 nmol/site), alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA; 135 pmol/site), (+/-)-1-aminocyclopentane-trans-1,3-dicarboxylic acid (trans-ACPD; 50 nmol/site) and substance P (SP, 135 ng/site). Tramadol injected by the i.t. route (25-400 nmol/site) also produced inhibition of glutamate-, AMPA , trans-ACPD- and SP-induced biting response. Pretreatment with tramadol by the i.p. and i.t. routes had no significant effect against the kainate- and N-methyl D-aspartic acid (NMDA)-mediated biting response in mice. PMID- 20016247 TI - Validation of cyanoacrylate method for collection of stratum corneum in human skin for lipid analysis. AB - BACKGROUND AND OBJECTIVE: Lipids in the stratum corneum (SC) are of major importance for the skin barrier function. Many different methods have been used for the collection of SC for the analysis of SC lipids. The objective of the present study was to validate the cyanoacrylate method for the collection of SC in relation to lipid analysis. METHODS: The results of the lipid analysis (ceramide/cholesterol and ceramide profile) of SC samples obtained by the cyanoacrylate method were compared to the results of the lipid analysis of mechanically removed SC samples. The intra- and interindividual variations in lipid composition were assessed when using the cyanoacrylate method, and lipid compositions in cyanoacrylate samples and samples taken from different depths of SC were compared. RESULTS: No statistically significant differences were found between mean values of lipids from the mechanically removed total thickness of the SC and cyanoacrylate samples. With respect to the cyanoacrylate samples, the intraindividual variation was significantly smaller than the interindividual variation, and the results did not indicate a change in lipid profile related to the depth of SC. The results clearly indicate that the cyanoacrylate method used for obtaining SC for lipid analysis is a useful and valid method for the purpose. PMID- 20016248 TI - Different pathways are involved in arsenic-trioxide-induced cell proliferation and growth inhibition in human keratinocytes. AB - BACKGROUND: Arsenic is a carcinogen that is associated with an increased risk of human skin cancer. On the other hand, arsenic trioxide (As(2)O(3)) has potential anticancer activity against a wide range of carcinomas. The mechanisms involved in these two opposing processes remain unclear. METHODS: We used normal human keratinocytes (NHK), the human keratinocyte HaCaT cell line and human epidermal carcinoma cells (A431 cell line) to investigate potential pathways involved in the effects on cell proliferation and growth inhibition by different concentrations of As(2)O(3). RESULTS: At low concentrations (0.5-32 nM), As(2)O(3) enhanced keratinocyte proliferation and regulated the expression of about 172 genes. Among them, cell cycling pathway genes (including CDK4 and E2F1) were significantly upregulated. At high concentrations (0.5-10 microM), As(2)O(3) inhibited cell growth in NHK and HaCaT cells, but not in A431 cells. As(2)O(3) significantly induced NHK and HaCaT apoptosis through the activation of caspase 3, as well as cell cycle arrest at the G2-M phase. CONCLUSION: Our data suggest that different pathways are involved in As(2)O(3)-mediated proliferation and growth inhibition. In addition, skin carcinoma cells were resistant to As(2)O(3) induced cell growth inhibition and apoptosis when compared to NHK and HaCaT cells. Therefore, As(2)O(3) may not be appropriate for treatment of skin carcinomas. PMID- 20016249 TI - Effect of helium-neon laser irradiation on hair follicle growth cycle of Swiss albino mice. AB - We report the results of a study carried out to investigate the effect of helium neon (He-Ne) laser (632.8 nm) irradiation on the hair follicle growth cycle of testosterone-treated and untreated mice. Both histology and optical coherence tomography (OCT) were used for the measurement of hair follicle length and the relative percentage of hair follicles in different growth phases. A positive correlation (R = 0.96) was observed for the lengths of hair follicles measured by both methods. Further, the ratios of the lengths of hair follicles in the anagen and catagen phases obtained by both methods were nearly the same. However, the length of the hair follicles measured by both methods differed by a factor of 1.6, with histology showing smaller lengths. He-Ne laser irradiation (at approximately 1 J/cm(2)) of the skin of both the control and the testosterone treated mice was observed to lead to a significant increase (p < 0.05) in % anagen, indicating stimulation of hair growth. The study also demonstrates that OCT can be used to monitor the hair follicle growth cycle, and thus hair follicle disorders or treatment efficacy during alopecia. PMID- 20016250 TI - Bacterial burden of worn therapeutic silver textiles for neurodermitis patients and evaluation of efficacy of washing. AB - To reduce pruritus and colonization with Staphylococcus aureus, textiles containing silver are increasingly used as therapeutic option for patients with atopic dermatitis (AD). While wearing such textiles, the contained silver is in close contact with the patient's skin. The silver serves two purposes: to reduce bacterial colonization of the skin, and to prevent contamination of the textile with ensuing growth of microorganisms. It is unknown whether the silver impregnation is able to reduce bacterial contamination of the textile during wearing and to prevent bacterial growth within the textile. The aim of this study was to investigate the bacterial contamination in textiles containing silver versus placebo worn by patients with AD and to determine the efficacy of processing worn textiles by manual and machine-based washing. Additionally, the effect of silver textiles on S. aureus and total bacterial counts colonizing the skin of AD patients was analyzed. The reduction factor of silver textile compared to placebo was 0.5 log steps against S. aureus and 0.4 log steps against total bacteria. Silver textiles exhibited significantly less S. aureus as well as total bacterial colonization after 2 days of wearing without washing, as compared with a placebo textile. On placebo textiles 385.6 +/- 63.5 CFU total bacteria and 236.5 +/- 49.9 CFU S. aureus, and on silver textiles 279.9 +/- 78.7 CFU total bacteria and 119.3 +/- 39.4 CFU S. aureus were found on the inner side of the textiles facing the neurodermitis lesions. However, the unexpectedly high residual contamination despite the silver exposure represents a potential risk as recontamination source of S. aureus that could maintain the proinflammatory process in AD. This contamination is nearly completely eliminated by machine based washing at 60 degrees C using conventional washing powder. AD patients wearing silver textiles should change their used clothes at least daily and wash them in a washing machine at 60 degrees C. PMID- 20016252 TI - The Phenion full-thickness skin model for percutaneous absorption testing. AB - In recent years many efforts have been made to replace dermal toxicity testing of chemicals in the animal by in vitro assays. As a member of a German research consortium, we have previously contributed to the validation of an in vitro test protocol for percutaneous absorption studies on the basis of reconstructed human epidermis and both human and pig skin ex vivo. Aiming to assess the barrier properties of a newly developed reconstructed skin model, this protocol has now been transferred to the Phenion Full-Thickness Skin Model (FT model). The permeation of testosterone and caffeine was quantified in parallel to that of pig skin using Franz-type diffusion cells. In addition, the permeation of benzoic acid and nicotine was studied. As expected, the FT model is more permeable than pig skin, yet its barrier properties are well in accordance with those of reconstructed human epidermis when compared to previous data. In fact, the FT model most efficiently retards testosterone as the compound of highest lipophilicity, which can be explained by an additional uptake by a reservoir formed by the dermis equivalent. Thus, the structure closely parallels human skin. In consequence, the Phenion FT model appears to be suitable for percutaneous absorption studies in hazard analysis and should be subjected to a catch-up validation study. PMID- 20016251 TI - Expression of Ca2+ -activated K+ channels in human dermal fibroblasts and their roles in apoptosis. AB - BACKGROUNDS/AIMS: Cell proliferation and apoptosis are responsible for maintaining normal tissue homeostasis, and K(+) currents play important roles in regulating the physiological balance between them. This function of Ca(2+) activated K(+) (K(Ca)) channels has been demonstrated in many types of tissues, but not in dermal fibroblasts. We investigated the expression of K(Ca) channels and their effects on proliferation and apoptosis in human dermal fibroblasts. METHODS: We used discoidin domain receptor 2 immunostaining to identify human dermal fibroblasts, and reverse transcription polymerase chain reaction, Western blot analysis and electrophysiological patch clamp recordings to evaluate the expression and characteristics of 3 members of the K(Ca) channel family, large conductance K(Ca) (BK), intermediate-conductance K(Ca) (IK) and small-conductance K(Ca) channels. We also used the 3-(4,5-dimethyl-2-yl)-2,5-diphenyltetrazolium bromide assay, flow cytometry, Hoechst 33258 staining and Depsipher staining to investigate the effects of K(Ca) channels on cell proliferation and the mechanisms involved. RESULTS AND CONCLUSIONS: All 3 members of the K(Ca) channel family were found in fibroblasts. 1,3-Dihydro-1-[2-hydroxy-5 (trifluoromethyl)phenyl]-5-(trifluoromethyl)-2H-benzimidazol-2-one (NS1619, a BK channel activator) or 1-ethyl-2-benzimidazolinone (EBIO, an IK channel activator) decreased the proliferation of fibroblasts and induced apoptotic changes by mitochondrial membrane potential disruption. However, a pan-caspase inhibitor (Z VAD-fmk) failed to prevent the apoptotic changes. Our findings indicate that 3 types of functional K(Ca) channels are expressed in human dermal fibroblasts and are involved in apoptosis of the cells through the mitochondrial apoptotic pathway, but seemingly in a caspase-independent manner. PMID- 20016253 TI - Pre-treatment with Aloe vera juice does not enhance the in vitro permeation of ketoprofen across skin. AB - BACKGROUND/AIMS: The potential of pre-treating skin with Aloe vera juice as a penetration enhancer was evaluated in vitro using ketoprofen as model permeant. METHODS: To excised porcine skin mounted in Franz diffusion cells was applied either: (1) commercial Aloe vera; (2) commercial Aloe vera followed by massaging; (3) previously boiled commercial Aloe vera; (4) water (negative control); (5) tea tree oil (positive control). After 1 h, the pre-treatment was removed and the skin dosed with a saturated solution of ketoprofen in polyethylene glycol 400; the appearance of drug in the receptor phase was then monitored by HPLC. RESULTS: No statistically significant differences in the transdermal delivery of ketoprofen were observed between water and all the Aloe vera pre-treatments (p > 0.05). The tea tree oil pre-treatment was significantly different to all others (p < 0.05). CONCLUSION: Aloe vera appears to have no value as a penetration enhancer when used as a pre-treatment, although the data indirectly support the mechanism of action proposed previously, work when used 'within-vehicle'. Handling household products containing Aloe vera appears not to leave the user at elevated risk of subsequent absorption of exogenous chemicals. PMID- 20016254 TI - The influence of Hfq and ribonucleases on the stability of the small non-coding RNA OxyS and its target rpoS in E. coli is growth phase dependent. AB - OxyS is one of at least three small non-coding RNAs, which affect rpoS expression. It is induced under oxidative stress and reduces the levels of the stationary phase sigma factor RpoS. We analyzed the turn-over of OxyS and rpoS mRNA in early exponential and in stationary growth phase in different E. coli strains to learn more about the mechanisms of processing and about a possible impact of processing on growth-dependent regulation. We could not attribute a major role of RNase E, RNase III, PNPase or RNase II on OxyS turn-over in exponential growth phase. Only the simultaneous lack of RNase E, PNPase and RNase II activity resulted in some stabilization of OxyS in exponential growth phase, implying the action of multiple ribonucleases on OxyS turn-over. A major role of RNase E on OxyS stability was observed in stationary phase and was dependent on the presence of the RNA binding protein Hfq and of DsrA, one of the other small RNAs binding to rpoS mRNA. Our data also confirm a role of RNase III in rpoS turn over, however, only in exponential growth phase.We conclude that OxyS and rpoS mRNA processing is influenced by different RNases and additional factors like Hfq and DsrA and that the impact of these factors is strongly dependent on growth phase. PMID- 20016255 TI - New connections identify Sch9 as a central node in ribosome biosynthesis. PMID- 20016256 TI - POPX adds to cell motility. PMID- 20016257 TI - The specific roles of mitotic cyclins revealed. PMID- 20016258 TI - Real time with caspase-2. PMID- 20016259 TI - Interaction between Rae1 and cohesin subunit SMC1 is required for proper spindle formation. PMID- 20016260 TI - Cdc48 connects with eIF3. PMID- 20016261 TI - A novel role for Chk2 after DNA damage in mitosis? PMID- 20016262 TI - Cell death or survival: the complex choice of the retinoblastoma tumor suppressor protein. PMID- 20016263 TI - Cell polarity, Notch signaling and neurogenesis. PMID- 20016264 TI - Mechanism of MAPK-targeted gene expression unraveled in plants. PMID- 20016265 TI - Wnt signaling and specification of the respiratory endoderm. PMID- 20016266 TI - Emerging role of FKBP51 in AKT kinase/protein kinase B signaling. PMID- 20016267 TI - A novel parkin-mediated transcriptional function links p53 to familial Parkinson's disease. PMID- 20016268 TI - TOS1 is circularly permuted 1,3-beta-glucanase. PMID- 20016269 TI - Isogenic mutant human cells: a new tool for personalized cancer medicine. PMID- 20016270 TI - The long unwinding road: XPB and XPD helicases in damaged DNA opening. AB - The mammalian nucleotide excision repair (NER) pathway removes dangerous bulky adducts from genomic DNA. Failure to eliminate these lesions can lead to oncogenesis, developmental abnormalities and accelerated ageing. TFIIH is a central NER factor that opens the damaged DNA through the action of its two helicases (XPB and XPD) prior to incision. Here we review our recently published data that suggest specific and distinct roles for these two helicases in NER. We also discuss the regulation of XPB and XPD enzymatic activities within TFIIH and repair complexes, and show that mutations impeding enzyme-regulator interaction contribute to genetic disorders. Understanding the fundamental molecular mechanism regulating NER is a crucial aspect of cancer therapy since the resistance to chemotherapy treatment relies on the capacities of the cell to eliminate drug-induced DNA lesions. PMID- 20016271 TI - Inhibition of cancer progression by rifampicin: involvement of antiangiogenic and anti-tumor effects. AB - Very recently, we reported that an antituberculous agent, rifampicin, exerts potent inhibitory effects on hepatic tumors when administered orally at low doses. The in vitro effects of rifampicin to rapidly downregulate angiogenesis and mitogenesis-related genes in cultured endothelial cells are reminiscent of endostatin, one of the most well-studied angiogenesis inhibitors. However, rifampicin at the expected hepatobiliary concentrations after low oral doses showed more complete antiproliferative effects on endothelial cells, which make rifampicin favorable as an adjunct anticancer regimen. Since rifampicin undergoes hepatic accumulation resulting from the enterohepatic circulation, it may be especially beneficial for targeting hepatobiliary tumors. In the present paper, we extend our observations on the antiangiogenic properties of rifampicin and further elaborate on its direct antitumor effects on a variety of human cancer derived cells. PMID- 20016272 TI - Genome wide identification of Plasmodium falciparum helicases: a comparison with human host. AB - Helicases are enzymes which catalyze the unwinding of nucleic acid substrate in an energy-dependent manner. These are characterized by the presence of nine well defined conserved motifs and are essential for almost all the processes involving nucleic acids. Plasmodium falciparum causes the most virulent form of malaria. The control of malaria is becoming complicated due to the spread of resistance of both the mosquito vector and the parasite to insecticides and anti-malarial drugs. Helicases could be used as feasible drug target for control of malaria. The P. falciparum genome is completely sequenced but the annotation is still in progress. To identify members of various well defined helicase families, I used the bioinformatics approach and helicase domain sequences to search the P. falciparum genome sequence database. In addition to the homologues for a number of human helicases, some novel parasite specific helicases were also identified. I describe the members of DEAD-box, DEAH box, RuvB, Superkiller family, RecQ and repair helicases from P. falciparum. The detailed studies of these helicases will help in identifying a specific enzyme, which could be used as potential target to control the replication and transmission of the malaria parasite. PMID- 20016273 TI - Opening the DNA repair toolbox: localization of DNA double strand breaks to the nuclear periphery. AB - Efficient repair of DNA double strand breaks is essential for cells to avoid increased mutation rates, genomic instability, and even cell death. Consequently, cells have evolved multiple mechanisms for rapidly repairing these DNA lesions, including error-free homologous recombination as well as error-prone pathways such as nonhomologous end joining. What happens to DSBs that are repaired inefficiently or not at all? Recently, several studies in budding yeast have shown that these more recalcitrant DSBs are localized to the nuclear periphery through interactions between the nuclear envelope protein, Mps3, and proteins associated with DSB chromatin. Why these DSBs are tethered to the nuclear periphery is still not clear, though the current view is that alternative repair pathways may be activated at the periphery in a final attempt to repair the lesion. In this Extra View, we discuss these recent reports, and we show that the Est1 component of the telomerase machinery plays an essential role in anchoring DSB chromatin to the nuclear envelope protein, Mps3. PMID- 20016275 TI - Yes-associated protein 1 regulates the stability of DeltaNp63alpha. AB - Regulation of DeltaNp63alpha can be achieved at the transcriptional and post translational levels, with the latter being greatly influenced by external stimuli such as DNA damaging agents. In this present study, we demonstrated that DeltaNp63alpha physical interacts with Yes-associated protein 1 (Yap1). Ectopic expression of Yap1 decreased the stability and the half-like of DeltaNp63alpha and resulted in ubiquitin-dependent degradation of DeltaNp63alpha. Further, the cisplatin mediated degradation of DeltaNp63alpha is attenuated with the downregulation of Yap1. Thus, our results strongly suggest that Yap1 plays a role in the regulation of endogenous DeltaNp63alpha levels and is likely to contribute to the regulation of DeltaNp63alpha, in physiological conditions. PMID- 20016274 TI - TERRA, CpG methylation and telomere heterochromatin: lessons from ICF syndrome cells. AB - Self-reinforcing negative feedback loops are commonly observed in biological systems. RNA-mediated negative feedback loops have been described in the formation of heterochromatin at centromeres in fission yeast and the inactive X chromosome in mammalian cells. The telomere repeat-containing RNA (TERRA) has also been implicated in the formation of telomeric heterochromatin through a self reinforcing negative feedback loop. In cells derived from human ICF syndrome, TERRA levels are abnormally elevated and telomeres are abnormally shortened. We now show that telomere heterochromatin is also abnormal in ICF cells. We propose that ICF cells fail to reinforce the TERRA-dependent negative feedback loop as a result of the inability to establish heterochromatin at subtelomeres. This failure is likely due to the lack of DNMT3b and DNA methylation, which is a genetic lesion associated with ICF syndrome. Failure of this feedback mechanism leads to catastrophic telomere dysfunction and chromosome instability. PMID- 20016276 TI - During hypoxia, HUMMR joins the mitochondrial dance. AB - Mitochondrial distribution is integrally related to cellular function. Highly polarized cells, such as neurons, likely depend on mitochondrial transport to maintain proper synaptic function and neurite plasticity. In some cases, mitochondrial transport is also required for cellular migration and proper calcium signaling in non-neuronal cells. Over the past few years, much progress has been made in identifying proteins that control mitochondrial transport and distribution. Miro and Milton, which are two outer mitochondrial membrane proteins, tether mitochondria to kinesin motor proteins. Our recent work identified a novel protein, HUMMR, which interacts with Miro. While present in normoxia, HUMMR protein abundance is markedly induced by hypoxia through a HIF-1 dependent mechanism. Knock down of HUMMR function diminishes the number of mitochondria in the axon, an effect that was more prominent in neurons exposed to hypoxia. Interestingly, in hypoxic neurons, knock down of HUMMR also diminished the number of anterograde moving mitochondria, but increased the number of retrograde moving mitochondria. Thus, HUMMR is a protein which biases mitochondrial movement in the anterograde direction in response to hypoxia. The implication for this biased transport of mitochondria during hypoxia on neuronal function and viability is yet to be discerned. Regardless, since hypoxia is prominent during ischemia and in solid tumors, HUMMR likely contributes to mitochondrial distribution under these conditions. As such, HUMMR may influence cellular function that is dependent upon the correct mitochondrial localization. PMID- 20016277 TI - The multiple roles of Bub1 in chromosome segregation during mitosis and meiosis. AB - Aneuploidy, any deviation from an exact multiple of the haploid number of chromosomes, is a common occurrence in cancer and represents the most frequent chromosomal disorder in newborns. Eukaryotes have evolved mechanisms to assure the fidelity of chromosome segregation during cell division that include a multiplicity of checks and controls. One of the main cell division control mechanisms is the spindle assembly checkpoint (SAC) that monitors the proper attachment of chromosomes to spindle fibers and prevents anaphase until all kinetochores are properly attached. The mammalian SAC is composed of at least 14 evolutionary-conserved proteins that work in a coordinated fashion to monitor the establishment of amphitelic attachment of all chromosomes before allowing cell division to occur. Among the SAC proteins, the budding uninhibited by benzimidazole protein 1 (Bub1), is a highly conserved protein of prominent importance for the proper functioning of the SAC. Studies have revealed many roles for Bub1 in both mitosis and meiosis, including the localization of other SAC proteins to the kinetochore, SAC signaling, metaphase congression and the protection of sister chromatid cohesion. Recent data show striking sex specific differences in the response of germ cells to alterations in Bub1 activity. Proper Bub1 functioning is particularly important during oogenesis in preventing the generation of aneuploid gametes that can have detrimental effects on the health status of the fetus and the newborn. These data suggest that Bub1 is a master regulator of SAC and chromosomal segregation in both mitosis and meiosis. Elucidating its many essential functions in regulating proper chromosome segregation can have important consequences for preventing tumorigenesis and developmental abnormalities. PMID- 20016278 TI - Ligand-driven activation of the notch pathway in T-ALL and solid tumors: why Not(ch)? AB - The Notch pathway is an evolutionally conserved cell-cell interaction signalling system involved in several key aspects of cell life, ranging from differentiation and proliferation to apoptosis. As such, it plays an important role in development, homeostasis, angiogenesis and various diseases. Over-activation of the Notch pathway has often been reported in cancer, leading to a variety of effects including increased proliferation, protection from apoptosis and maintenance of cancer initiating cells. Additionally, this signalling pathway has also been involved in tumor angiogenesis. The clearest example of oncogenic Notch signalling is observed in T acute lymphoblastic leukemia (T-ALL), an aggressive neoplasm of immature T-cells, due to genetic alterations leading to ligand independent increased Notch1 receptor signalling. In solid tumors, however, extrinsic regulation through canonical cell-cell interactions appears to drive activation of the pathway. We recently found that triggering of Notch3 signalling by DLL4 expressed on angiogenic endothelial cells promotes escape of T-ALL cells from tumor dormancy in a xenograft model. This observation discloses un unsuspected role for ligand-dependent regulation of Notch receptors in T-ALL cells, suggesting that blocking extrinsic Notch activation by anti-DLL4 or other ligand-targeted drugs could represent a novel therapeutic approach for this aggressive malignancy. PMID- 20016279 TI - Tumor suppression by ARF: gatekeeper and caretaker. AB - ARF is a vital tumor suppressor and its loss contributes significantly to cancer. The frequency in which ARF is mutated, deleted or silenced is second to the loss of p53. The most documented and widely accepted activity of ARF is mediated through its activation of the p53 transcriptional program by inhibiting MDM2 function. However, several lines of evidence have surfaced demonstrating that ARF possesses p53-independent functions. One of these p53-independent functions is ARF's regulation of the E2F family. The E2F/DP transcription factor is critical for cell cycle progression. The balance between activator and repressor E2Fs regulates the expression of E2F target genes and thus cell proliferation as well as other cellular functions such as checkpoint, chromosome assembly and repair. Through its ability to bind directly to DP1, ARF can cause dissociation of both activator and repressor E2Fs. While the regulation of the activator E2Fs is related to cell cycle arrest, there is evidence that the regulation of the repressors, E2F4 and E2F5, is significant in maintaining genomic stability. PMID- 20016280 TI - The roles of nitric oxide synthase and eIF2alpha kinases in regulation of cell cycle upon UVB-irradiation. AB - In response to ultraviolet light (UV)-induced damage, cells initiate cellular recovery mechanisms including activation of repair genes and redistribution of cell cycle phases. While most studies have focused on DNA damage-inducible transcriptional regulation of cell cycle checkpoints, translational regulation also plays an important role in control of cell cycle progression upon UV irradiation. UV-irradiation activates two kinases, PERK and GCN2, which phosphorylate the alpha subunit of eukaryotic initiation factor 2 (eIF2alpha) and subsequently inhibit protein synthesis. We recently identified an upstream regulator, nitric oxide synthase (NOS), which controls the activation of both PERK and GCN2 upon UVB-irradiation. Our data suggested that UVB induces NOS activation and NO(.) production, which reacts with superoxide (O(2)(*-)) to form peroxynitrite (ONOO(-)) and activate PERK. The NO(*) production also leads to L Arg depletion and GCN2 activation. The elevation of nitric oxide and activation of PERK/GCN2 have been shown to play roles in regulation of cell cycle upon UVB irradiation. In the present study, we show that the cell cycle phases were redistributed by inhibition of NOS activation or reduction of oxidative stress upon UVB irradiation, indicating the roles of NO(*) and its oxidative products in regulation of cell cycle. We also demonstrate that both PERK and GCN2 were involved in regulation of cell cycle upon UVB-irradiation, but the regulation is independent of eIF2alpha phosphorylation. While the mechanism for UVB-induced cell cycle control is yet to be unraveled, we here discuss the differential roles of NOS, PERK and GCN2 in regulation of cell cycle upon UVB-irradiation. PMID- 20016281 TI - Int6 and Moe1 interact with Cdc48 to regulate ERAD and proper chromosome segregation. AB - Int6/eIF3e is implicated in tumorigenesis, but its molecular functions remain unclear. We have studied its fission yeast homolog Yin6, reporting that it regulates proteolysis by controlling the assembly/localization of proteasomes, and binds directly to another conserved protein, Moe1. In the present study, we isolated Cdc48 as a Moe1-binding protein from a yeast two-hybrid screen, and confirmed biochemically that they form a stable complex in fission yeast. Overexpressing Moe1 or Yin6 partially rescued phenotypes of cdc48 mutants; conversely, overexpressing Cdc48 partially rescued phenotypes of moe1 or yin6 mutants. Mutants defective in both Cdc48 and the Yin6-Moe1 complex showed growth defects that were far more severe than either alone. These double mutants were severely deficient in endoplasmic reticulum associated degradation (ERAD), as they were hypersensitive to accumulation of misfolded proteins. In addition, their chromosomes showed frequent defects in spindle attachment and segregation- these mitotic defects correlated with Ase1 and Bir1/survivin mislocalization. These results suggest that Cdc48, Yin6 and Moe1 act in the same protein complex to concertedly control ERAD and chromosome segregation. Many of these properties are evolutionarily conserved in humans, since human Cdc48 rescued the lethality of the yeast cdc48Delta mutant, and Int6 and Moe1/eIF3d bind Cdc48 in human cells. PMID- 20016282 TI - Vitamin D: Proteases, protease inhibitors and cancer. AB - The active vitamin D metabolite 1alpha,25-dihydroxyvitamin D(3) (1,25(OH)(2)D(3), Calcitriol) is a major regulator of gene expression in higher organisms. Protein abundance is an endpoint of gene expression that results from the balance between induction and degradation and is essential for adequate cell function. Proteins are degraded by proteases whose activity is in turn controlled by a number of endogenous protease inhibitors. 1,25(OH)(2)D(3) regulates several proteases and protease inhibitors in different cell types, putatively contributing to its regulatory effects of cell physiology. We have recently shown that 1,25(OH)(2)D(3) strongly induces the expression of cystatin D, an inhibitor of several cysteine proteases of the cathepsin family. Cystatin D induction may contribute to the antitumor effect of 1,25(OH)(2)D(3) against colon cancer by mechanisms that are both dependent and independent of cathepsin inhibition. Transcriptomic studies suggest that 1,25(OH)(2)D(3) also modulates the function of the ubiquitin-proteasome system. Thus, proteases and protease inhibitors are candidates to mediate to a certain extent the complex action of 1,25(OH)(2)D(3) in cancer cells. PMID- 20016283 TI - Processing of O6-methylguanine into DNA double-strand breaks requires two rounds of replication whereas apoptosis is also induced in subsequent cell cycles. AB - The DNA adduct O(6)-methylguanine (O(6)MeG) induced by environmental genotoxins and anticancer drugs is a highly mutagenic, genotoxic and apoptotic lesion. Apoptosis induced by O(6)MeG requires mismatch repair (MMR) and proliferation. Models of O(6)MeG-triggered cell death postulate that O(6)MeG/T mispairs activate MMR giving rise to either direct genotoxic signaling or secondary lesions that trigger apoptotic signaling in the 2(nd) replication cycle. To test these hypotheses, we used a highly synchronized cell system competent and deficient for the repair of O(6)MeG adducts, which were induced by the S(N)1 methylating agent N-methyl-N'-nitro-N-nitrosoguanidine (MNNG). We show that DNA double-strand breaks (DSBs) are formed in response to O(6)MeG at high level in the 2nd S/G(2) phase of the cell cycle. This is accompanied by ATR and Chk1 phosphorylation, G(2)/M arrest and late caspase activation. Although cells undergo apoptosis out of the 2nd G(2)/M-phase, the majority of them recovers and undergoes apoptosis after passing through additional replication cycles. The late apoptotic effects were completely abolished by O(6)-methylguanine-DNA methyltransferase, indicating that non-repaired O(6)MeG is carried over into subsequent generations, eliciting there a late apoptotic response. We also demonstrate that with a low, non-toxic dose of MNNG the passage of cells through the 1st and 2nd S-phase is not delayed, although the dose is able to induce excessive sister chromatid exchanges. This suggests that a significant amount of O(6)MeG can be tolerated by recombination, which is a fast and highly efficient process preventing from S-phase blockage, DSB formation and cell death. PMID- 20016284 TI - The Drosophila retinoblastoma protein induces apoptosis in proliferating but not in post-mitotic cells. AB - The retinoblastoma protein, pRb, plays important roles in many processes implicated in cell fate decisions, including cell cycle, differentiation and apoptosis. In cell cycle regulation, pRb interacts principally with the E2F transcription factor family members to inhibit the transcription of many genes controlling cell cycle progression. In this study, we focused on the role of pRb in apoptosis, which is much less clear than its role in cell cycle regulation. Indeed, pRb has been found to be either pro- or anti-apoptotic. To clarify how the proliferative status of the cells impacts the role of pRb in apoptosis, we used Drosophila to induce RBF (the pRb fly homologue) expression in different cellular and developmental contexts. We found that RBF expression induces apoptosis in different proliferative tissues in a caspase-dependent manner, whereas this effect was not observed in differentiated post-mitotic cells. Furthermore, RBF-induced apoptosis in proliferating cells was inhibited by co expression of dE2F1, an antagonistic partner of RBF in cell cycle regulation. These results are in agreement with the view that the apoptotic properties of pRb are tightly linked to, and are probably a consequence of, an effect on cell cycle progression. Moreover, we show for the first time that RBF has a direct anti apoptotic effect on Dmp53-induced cell death in post-mitotic cells only. Taken together, these data clearly show that RBF can exert a dual role in the control of apoptotic processes, and that its properties depend on the proliferative status of the cells. PMID- 20016285 TI - Single cell nanoparticle tracking to model cell cycle dynamics and compartmental inheritance. AB - Single cell encoding with quantum dots as live cell optical tracers for deriving proliferation parameters has been developed using modelling to investigate cell cycle and proliferative outputs of human osteosarcoma cells undergoing mitotic bypass and endocycle routing. A computer-based simulation of the evolving cell population provides information on the dilution and segregation of nanoparticle dose cell by cell division and allows quantitative assessment of patterns of division, at both single cell and including whole population level cell cycle routing, with no a-priori knowledge of the population proliferation potential. The output therefore provides a unique mitotic distribution function that represents a convolution of cell cycle kinetics (cell division) and the partitioning coefficient for the labelled cell compartment (daughter-daughter inheritance or lineage asymmetry). The current study has shown that the cellular quantum dot fluorescence reduced over time as the particles were diluted by the process of cell division and had the properties of a non-random highly asymmetric event. Asymmetric nanoparticle segregation in the endosomal compartment has major implications on cell-fate determining signaling pathways and could lead to an understanding of the origins of unique proliferation and drug-resistance characteristics within a tumour cell lineage. PMID- 20016286 TI - The POPX2 phosphatase regulates cancer cell motility and invasiveness. AB - Rho GTP ases play major roles in the regulation of actin cytoskeleton, cell movement and cell cycle. PAK, one of the effector kinases of these small GTP ases, has long been associated with different types of cancer. Therefore, it is likely that deregulation of PAK activity or expression may contribute to the development of cancer. POP X2, a PP 2C serine/threonine phosphatase, is known to dephosphorylate PAK and downregulate its activity. We find that POPX2 is expressed in a wide variety of tumour cell lines, the levels being highest in the more invasive MDA-MB-231 and lowest in the non-invasive MCF7 breast cancer lines. We show that silencing of POPX2 reduces the amount of stress fibers and focal adhesions in both cells lines. Interestingly, POPX2 deficiency dramatically reduces cell motility and invasiveness in MDA-MB-231 cells, and cell motility in MCF7 cells. Conversely, overexpression of POP X2 in MDA-MB-231 and MCF7 cells increased their motility. The silencing of POP X2 also inhibits the expression of beta1 integrin implying that POP X2 may modulate cell attachment to the extra cellular matrix, as reflected in diminished initial colonization of POPX2 knockdown cells in nude mice. Based on these results, we propose a mechanism by which POP X2 regulates the invasive behavior of the cells. PMID- 20016287 TI - Metformin extends life span of HER-2/neu transgenic mice and in combination with melatonin inhibits growth of transplantable tumors in vivo. AB - Population studies have shown that treatment with the antidiabetic biguanide metformin significantly reduced cancer risk. In our animal studies, metformin delayed the onset of mammary adenocarcinoma (MAC) in transgenic HER-2/neu mice but not the onset of spontaneous mammary tumors in female SHR mice. Pineal hormone also inhibits mammary carcinoma development in HER2/neu transgenic mice as well as in female SHR mice. Here we demonstrated that a combination of metformin and melatonin significantly inhibits growth of transplanted tumors in mice. Metformin (0.5 mg/ml in drinking water) increased mean life span by 8% and MAC latency by 13.2% (p < 0.05) in HER2/neu mice. The treatment with melatonin alone (2 mg/L in drinking water during the night time) or combined treatment with metformin (0.5 mg/ ml in drinking water during the day time) + melatonin (2 mg/L in drinking water during the night time) did not influence mammary carcinogenesis in the mice. The treatment metformin alone inhibited the growth of transplantable HER2 mammary carcinoma in FVB/N male mice by 46% at the 45(th) day after transplantation (p < 0.001). The combined treatment with metformin + melatonin significantly suppressed Ehrlich tumor growth (by 40%, p < 0.001). These results suggest that metformin may be useful in prevention and treatment of breast cancer. PMID- 20016288 TI - Interferon-alpha2b induces p21cip1/waf1 degradation and cell proliferation in HeLa cells. AB - Type I interferons (IFNs) are a family of cytokines that exhibit various biological activities. Besides their roles in immune response, IFNs have been known to modulate cell proliferation and to induce apoptosis. Thus, IFNs are used as an antitumor agent against certain types of cancer, but it is unclear why many other cancers are not influenced by IFNs. Here, we found that IFN-alpha2b, a subfamily of IFN-alpha, enhanced proliferation of HeLa cells, a cell line derived from human cervical cancer. IFN-alpha2b was rather inhibitory on the growth of other types of cervical cancer cells including those positive for HPV. Among the proliferation- and the apoptosis-related genes, p21(cip1/waf1) (p21) was upregulated by IFN-alpha2b, whereas p53, p27 or BCL-2 associated X protein (BAX) was not affected. IFN-alpha2b did not alter promoter activities of p21 but did prolong the decay of p21 mRNA. In contrast, the level of p21 protein was lowered by IFN-alpha2b, and half-life analysis of p21 protein revealed that IFN-alpha2b enhances p21 protein instability in HeLa cells. Pretreatment of the cells with MG132, a proteasome inhibitor, abolished the IFN-alpha2b-mediated p21 degradation, suggesting that IFN-alpha2b accelerated the ubiquitin-proteasome dependent degradation of p21. Consistent with these results, IFN-alpha2b increased S-phase cell cycle distribution in HeLa cells. In addition, IFN-alpha2b liberated the cells from G(1)-phase arrest by 5-fluorouracil (5-FU) and from G(2) phase arrest by paclitaxel. These results provide a novel role of Type I IFNs in cell cycle regulation and may define an importance of individualized IFN-based therapy against specific types of cancer. PMID- 20016289 TI - Induction of papillary carcinoma in human ovarian surface epithelial cells using combined genetic elements and peritoneal microenvironment. AB - Papillary differentiation is one of the most common histological features of ovarian cancer, although the underlying mechanism that leads to such differentiation is not known. We hypothesized that human ovarian surface epithelial cells can be transformed into carcinoma with papillary differentiation by overexpressing HER2/neu in these cells. Mice were injected either subcutaneously or intraperitoneally with two immortalized human ovarian surface epithelial cell lines after enforced expression of HER-2/neu. Mice subcutaneously injected with tumor cells from either the T29Nt or T80Nt developed undifferentiated carcinomas. In contrast, mice injected intraperitoneally with T29Nt cells developed papillary carcinoma, and those injected intraperitoneally with T80Nt cells developed undifferentiated carcinoma. Our results demonstrate that ovarian surface epithelial cells can develop into papillary carcinoma in mice, and that the induction of papillary differentiation depends not only on specific genetic modifications but also on the tumor microenvironment and epithelial cell type from ovary from different patients. PMID- 20016290 TI - HIF-1alpha dysfunction in diabetes. AB - Diabetic wounds are a significant public health burden, with slow or nonhealing diabetic foot ulcers representing the leading cause of non-traumatic lower limb amputation in developed countries. These wounds heal poorly as a result of compromised blood vessel formation in response to ischemia. We have recently shown that this impairment in neovascularization results from a high glucose induced defect in transactivation of hypoxia-inducible factor-1alpha (HIF 1alpha), the transcription factor regulating vascular endothelial growth factor (VEGF) expression. HIF-1 dysfunction is the end result of reactive oxygen species induced modification of its coactivator p300 by the glycolytic metabolite methylglyoxal. Use of the iron chelator-antioxidant deferoxamine (DFO) reversed these effects and normalized healing of humanized diabetic wounds in mice. Here, we present additional data demonstrating that HIF-1alpha activity, not stability, is impaired in the high glucose environment. We demonstrate that high glucose induced impairments in HIF-1alpha transactivation persist even in the setting of constitutive HIF-1alpha protein overexpression. Further, we show that high glucose-induced hydroxylation of the C-terminal transactivation domain of HIF 1alpha (the primary pathway regulating HIF-1alpha/p300 binding) does not alter HIF-1alpha activity. We extend our study of DFO's therapeutic efficacy in the treatment of impaired wound healing by demonstrating improvements in tissue viability in diabetic mice with DFO-induced increases in VEGF expression and vascular proliferation. Since DFO has been in clinical use for decades, the potential of this drug to treat a variety of ischemic conditions in humans can be evaluated relatively quickly. PMID- 20016292 TI - Role of exercise in the prevention and management of lymphedema after breast cancer. PMID- 20016293 TI - Neural correlates of motor learning, transfer of learning, and learning to learn. AB - Recent studies on the neural bases of sensorimotor adaptation demonstrate that the cerebellar and striatal thalamocortical pathways contribute to early learning. Transfer of learning involves a reduction in the contribution of early learning networks and increased reliance on the cerebellum. The neural correlates of learning to learn remain to be determined but likely involve enhanced functioning of the general aspects of early learning. PMID- 20016294 TI - Skeletal muscle injury versus adaptation with aging: novel insights on perplexing paradigms. AB - A growing body of data supports a view that skeletal muscle's response after mechanical loading does not always result in the classically reported "injury response." Furthermore, current evidence supports a model of muscle adaptation and/or maladaptation, distinct from overt injury, in which myofiber degeneration and inflammation do not contribute as significantly as once reported even in aged populations. PMID- 20016295 TI - Balancing lymphedema risk: exercise versus deconditioning for breast cancer survivors. AB - Lymphedema, a common and feared negative effect of breast cancer treatment, is generally described by arm swelling and dysfunction. Risk averse clinical recommendations guided survivors to avoid the use of the affected arm. This may lead to deconditioning and, ironically, the very outcome women seek to avoid. Recently published studies run counter to these guidelines. PMID- 20016296 TI - Physical activity and hormonal regulation of appetite: sex differences and weight control. AB - Physical activity is an important contributor to regulation of energy balance and body composition. In this article, we separate the impact of exercise from the confounding influence of energy imbalance and highlight sex differences in hormonal and appetite responses to physical activity. The evolving story may influence our thinking regarding the use of physical activity to manage body composition. PMID- 20016297 TI - Sympathetic actions on the skeletal muscle. AB - The sympathetic nervous system (SNS) modulates several functions in skeletal muscle fibers, including metabolism, ionic transport across the membrane, and contractility. These actions, together with the sympathetic control of other organ systems, support intense motor activity. However, some SNS actions on skeletal muscles may not always be functionally advantageous. Implications for motor control and sport performance are discussed. PMID- 20016298 TI - Task-specific sex differences in muscle fatigue: is there a common underlying cause? PMID- 20016300 TI - Quadriceps inhibition may not be a predecessor of posttraumatic osteoarthritis associated with ACL injury. PMID- 20016302 TI - Looking forward with a grateful heart. PMID- 20016303 TI - Role of caveolin and heat shock protein 70 interaction in the antioxidative effects of an angiotensin II type 1 receptor blocker in spontaneously hypertensive rats proximal tubules. PMID- 20016304 TI - Spironolactone for all hypertensive patients? PMID- 20016305 TI - Enhance MSSE's online presence. PMID- 20016306 TI - Midportion Achilles tendinopathy--a cardiovascular disease? PMID- 20016308 TI - Polycystic ovary syndrome and estrogen metabolism in female athletes. PMID- 20016311 TI - If the shoe fits...don't get too comfortable! PMID- 20016312 TI - Kidney disease: easing pain at end of life. PMID- 20016314 TI - Blame the name for a med error? PMID- 20016318 TI - Caring for a patient after amputation. PMID- 20016320 TI - Making the connection. PMID- 20016321 TI - Guillain-Barre syndrome: a nursing nightmare that usually ends well. PMID- 20016323 TI - Uterine cancer. PMID- 20016324 TI - Understanding acute pancreatitis. PMID- 20016326 TI - Good news about writing for your local newspaper. PMID- 20016327 TI - Get the lowdown on ankle and foot ulcers. PMID- 20016328 TI - Caring for a patient with schizophrenia in a med-surg unit. PMID- 20016329 TI - Obstructed tracheostomy tubes: clearing the air. PMID- 20016330 TI - Managing pulmonary arterial hypertension: a heartwarming success story. PMID- 20016331 TI - Flushing out the truth about diabetes insipidus. PMID- 20016332 TI - Speaking up for older patients with hearing loss. PMID- 20016333 TI - Avoiding the sting of malaria. PMID- 20016334 TI - Where has all the meperidine gone? PMID- 20016335 TI - Documenting a patient's violent behavior. PMID- 20016336 TI - Insights on death & dying. PMID- 20016337 TI - Combination drugs... PMID- 20016338 TI - Acute dehydration. PMID- 20016340 TI - NAON Foundation ... building the future of orthopaedic nursing. PMID- 20016339 TI - With change comes great opportunities. PMID- 20016342 TI - Adhesive capsulitis: a case study. AB - Adhesive capsulitis (AC), also known as frozen shoulder, is a common and painful musculoskeletal disorder. The shoulder is a complex joint comprising 3 bones and multiple muscle groups, ligaments, and tendons. Accompanying the inflammation of the joint is stiffness that greatly restricts the patient's motion and strength. AC is a treatable condition requiring months of daily physical therapy. The reports that 1 in 7 Americans has a musculoskeletal impairment including shoulder problems that accounted for 14 million visits to physicians' offices in 2003. It is imperative that practitioners accurately assess and diagnose the patient with shoulder pain to facilitate an appropriate plan of care. This paper will discuss the case of a patient with AC including differential diagnoses, pathophysiology, and treatment options. PMID- 20016344 TI - Intraoperative neuromonitoring in pediatric spinal deformity surgery. AB - Pediatric spinal deformity includes a broad range of disorders with differing causes, natural histories, and treatments. Spinal deformity surgery is performed to arrest the progression of and correct coronal and sagittal plane deformities including scoliosis and kyphosis. It is also undertaken to improve cosmesis and decrease the pain that can be associated with certain types of spinal deformity. Iatrogenic spinal cord injury remains the most feared complication of corrective spine surgery. Intraoperative neuromonitoring may be the single innovation that has had the greatest impact in lowering neurological complication rates over the last 2 decades. It is currently recommended in pediatric spinal deformity surgery whenever cord-level spinal instrumentation is planned and reliable signals can be anticipated. This article will briefly discuss common types of pediatric spinal deformity and corrective spine surgery. The use of intraoperative neuromonitoring in pediatric spine surgery will also be reviewed. As our abilities to correct more complex spinal deformities continue to improve, the importance of more advanced methods to decrease the risk of intraoperative neurological complications will also continue to increase. PMID- 20016346 TI - Relationship between patient trust of nursing staff, postoperative pain, and discharge functional outcomes following a total knee arthroplasty. AB - PURPOSE: This correlational study examined the relationship between patient trust of nurses, level of postoperative pain, and discharge functional outcome following total knee arthroplasty. SAMPLE: The study included 68 participants. METHODS: Participants completed the Trust subscale of the Patient's Opinion of Nursing Care to assess trust of nursing staff. Numeric Analog Scale pain scores measured level of pain. Functional outcome was obtained from knee flexion gain scores. FINDINGS: Average trust score was 47.5, average Numeric Analog Scale pain score was 5.0, and average gain score was 26.8. Analysis suggested no statistically significant relationships between patient trust of staff nurses, level of postoperative pain, and discharge functional outcome. PMID- 20016347 TI - Relationship between dental caries and total joint arthroplasty at a Veterans Administration hospital. AB - It has previously been suspected that patients who undergo extensive dental procedures after having had a total joint arthroplasty (TJA) within the preceding 2 years may be at risk for seeding their TJA with infection if prophylaxis antibiotic coverage is not implemented one hour prior to the dental procedure. A review of the literature was performed to determine whether there may be other infections which may be accountable for TJA infections within the first 2 years following a TJA. Patients with systemic diseases, those who undergo extensive dental procedures, those with bladder or skin infection were found to be at risk for development of a TJA if prophylactic antibiotic therapy was not implemented in a timely manner. PMID- 20016348 TI - Mental status and surgical methods in patients with femoral neck fracture. AB - BACKGROUND: About one third of hip-fractured patients have dementia and thus may have difficulties adhering to postoperative instructions. Hip replacement is the most common treatment when a femoral neck fracture is displaced in healthy older people, whereas for those with dementia and other severe comorbidities, internal fixation (IF) is generally recommended. PURPOSE: To evaluate complications, functional outcome, and mortality for both surgical methods, IF and hemiarthroplasty (HAP), in older patients suffering from femoral neck fracture with or without dementia. SAMPLE: One hundred eighty patients, aged 70 years or older, who were operated on using IF (n = 69) in undisplaced femoral neck fracture and HAP (n = 111) if the fractures were displaced. DATA COLLECTION: Mental state was assessed using the Mini-Mental State Examination and Organic Brain Syndrome scale, and dementia and delirium were diagnosed according to the Diagnostic and Statistical Manual of Mental Disorders (Fourth Edition) criteria. Outcomes of mortality, complications, functional ability, and quality of life were measured. FINDINGS: There was no difference in complications or mortality at 4 months and 1 year for the IF or HAP groups. Patients with and without dementia, operated on with HAP, had a better functional outcome after 1 year than those operated on with IF. The result of this study indicates that dementia per se is not a reason for disqualifying those patients from the most appropriate surgical method. PMID- 20016349 TI - Ankle sprains and fractures in adults. AB - The orthopaedic nurse will undoubtedly care for patients with ankle injuries. Ankle sprains and fractures are common injuries seen in orthopaedic practices in the United States. To provide comprehensive nursing care of these injuries, nurses should understand ankle biomechanics as well as anatomy. Knowledge of fracture classification schemes, pathology of injury, and treatment modalities is also important. Fracture classification schemes vary as do treatment modalities. Ankle sprains and fractures can be treated operatively or nonoperatively, and orthopaedic nurses play an important role in the care of patients with such ankle injuries. PMID- 20016351 TI - Hip and knee arthroplasties. PMID- 20016353 TI - Analysis of parental and nurse weight estimates of children in the pediatric emergency department. AB - OBJECTIVE: To evaluate the accuracy of parent and triage nurse estimates of children's weights in the pediatric emergency department. METHODS: A convenience sample of parents was surveyed before triage regarding their child's chief complaint, when and where the child was last weighed, and an estimate of their child's weight. Children unable to be weighed or with possible dehydration or external orthopedic devices were excluded. Nurses also estimated children's weights before weighing them and were blinded to parent estimates. All nurses were experienced in the department, had participated in a triage course, and had completed a background survey. RESULTS: Nine hundred forty-two patients presenting to pediatric emergency department triage were enrolled, resulting in 812 surveys. Parents were more accurate at weight estimation than nurses, with 79% of parents and 83% of legal guardians estimating weights within 10%. Only 55% of nurse estimates were within 10%. Parents and nurses tended to underestimate patients' weights (P < 0.0001). Parent estimates were not affected by a child's age or sex or the location the child was last weighed (P > 0.05). Accuracy of nurse estimates was not related to nurse education or experience (P > 0.05). Parent estimates of weight were more accurate than nurse estimates 74.6% of the time. CONCLUSIONS: Parents were more accurate at estimating children's weights than triage nurses but were within 10% of the children's actual weights only 79% of the time. Nurse estimates were highly inaccurate. Other methods to estimate patient weights should be used when actual patient weights are unobtainable. PMID- 20016354 TI - Bruising in infants: those with a bruise may be abused. AB - Bruising in the young infant is rare, and if present, this may be a manifestation of physical child abuse. Early signs of abuse, such as bruising, are often overlooked or their significance goes unrecognized resulting in poor patient outcomes. In such cases, the opportunity to intervene and potentially prevent repeat injury is lost, and the child is placed back in harm's way. This brief report presents 3 cases of nonmobile infants who presented to health care providers with bruising before a subsequent fatal or near-fatal event. These cases emphasize the importance of including abusive trauma in the differential diagnosis of an infant with a bruise or a history of easy bruising and the importance of initiating a thorough trauma evaluation immediately and concomitantly with any other workup for the causes of bruising in the noncruising infant. PMID- 20016355 TI - Gastric artery apoplexy presenting as an acute abdomen in an adolescent. AB - Nontraumatic abdominal apoplexy, which is the occurrence of hemorrhage into the peritoneal cavity, is uncommon in the pediatric literature. Adult case reports of nontraumatic abdominal apoplexy include visceral artery aneurysms from the aorta, splenic, celiac, hepatic, gastric, and gastroepiploic arteries as well as gynecologic pathology. We identified 2 case reports of spontaneous abdominal apoplexy due to injury of the short gastric arteries in adult patients with histories of vomiting after alcohol consumption. This is the first reported case of nontraumatic abdominal apoplexy due to injury to the short gastric arteries in a pediatric patient. PMID- 20016356 TI - Atrial fibrillation after taser exposure in a previously healthy adolescent. AB - We are reporting a previously healthy adolescent who developed atrial fibrillation after being tased. He has a structurally normal heart on echocardiogram, normal electrolyte level and thyroid function test results, and a urine toxicology screen positive for marijuana. The patient ultimately required external defibrillation to convert his cardiac rhythm to normal sinus rhythm and has had no recurrent arrhythmias since hospital discharge (approximately 1 year). This is the first reported case of atrial fibrillation developing after a Taser shot, occurring in an adolescent without other risk factors. This case illustrates the arrhythmogenic potential of a Taser in otherwise healthy young individuals, and further study of occurrence of Taser-induced arrhythmias is warranted. PMID- 20016357 TI - Bedside ultrasound diagnosis of pulmonary contusion. AB - A 10-year-old boy presented to the emergency department after being struck by a van while crossing the street. He complained of right side chest pain, and a chest radiography was suggestive of pulmonary contusion. The treating physician performed a bedside ultrasound that revealed a right-sided pulmonary contusion that was subsequently confirmed on computed tomography of the thorax. The sonographic features of pulmonary contusion are described, and the possible role of lung sonography in the assessment of pediatric thoracic trauma is discussed. PMID- 20016358 TI - Spontaneous vaginal drainage of a pelvic abscess: an unusual presentation of perforated appendicitis. AB - We report a child with the unusual presentation of spontaneous vaginal drainage of a pelvic abscess after nonoperative management of perforated appendicitis. Although drainage through the rectum has been previously described, this is the first report of spontaneous transvaginal drainage of a pelvic abscess from appendicitis in a child. PMID- 20016359 TI - Hypophosphatemia-induced seizure in a child with diabetic ketoacidosis. AB - We report an unusual case of hypophosphatemia-related seizure in a child with diabetic ketoacidosis (DKA). A 1-year-old type 1 diabetic boy with hyperglycemia, ketoacidosis, and dehydration was admitted to the pediatric intensive care unit. After having received fluid replacement using isotonic solution with added potassium and continuous intravenous insulin administration according to the protocol for DKA, the patient was conscious, awake, and fed with breast milk. After 20 hours of pediatric intensive care unit stay, he presented 2 tonic-clonic seizures followed by apnea. One hour later, he had cardiorespiratory arrest, requiring cardiovascular support and mechanical ventilation. Serum phosphorus concentration was 1.0 mg/dL, and severe hypophosphatemia was diagnosed. Subsequent to intravenous phosphate replacement, he showed improved neurological and hemodynamic statuses. No other cause of cerebral complication was found. He had no neurologic lesions and was discharged. Although hypophosphatemia is a common complication of DKA treatment, phosphate supplementation has not been routinely recommended in the treatment of DKA. Early recognition and treatment of severe hypophosphatemia in the treatment of DKA are important to reduce the risk of neurological complications. PMID- 20016360 TI - Discitis and epidural abscess after dental extraction in a pediatric patient: a case report. AB - Neck pain is a relatively common pediatric complaint. Most is of benign etiology. We present an unusual case of epidural abscess with associated discitis in the cervical spine of a 12-year-old after a dental extraction. This potentially dangerous complication was diagnosed before neurological complications developed and was successfully treated with antibiotics alone, preventing the need for surgery. An extensive search of the literature has revealed only 5 cases of epidural abscess secondary to dental sepsis, and to the author's knowledge, this is the first reported case in the pediatric population. We present the case and a brief review of the literature on this subject. PMID- 20016361 TI - Pediatric emergency medicine: legal briefs. PMID- 20016362 TI - A 19-month-old girl with recurrent illness: over the hills and through the woods. PMID- 20016363 TI - ECGs in the ED. PMID- 20016365 TI - A phase II study of paclitaxel poliglumex in combination with transdermal estradiol for the treatment of metastatic castration-resistant prostate cancer after docetaxel chemotherapy. AB - Taxanes remain the only agents to extend survival in castration-resistant metastatic prostate cancer, but their impact on the natural history of this disease is modest. We sought to test the hypothesis that increased delivery of taxane chemotherapy to the tumor through the use of a macromolecular polymer-drug conjugate of paclitaxel modulated by estradiol could extend the utility of this class of drugs. Patients with metastatic adenocarcinoma of the prostate who progressed despite standard hormonal therapy and after docetaxel-containing chemotherapy were treated with transdermal estradiol (0.2 mg/24 h) for 4 weeks followed by the same dose of transdermal estradiol and paclitaxel poliglumex (PPX; 150 mg/m intravenous) every 28 days. The primary objective was to determine the level of activity of the regimen measured using a fraction of patients who experienced a confirmed decline in serum prostate-specific antigen (PSA) of 50% or more. A two-stage phase II study designed to identify a response rate of > or =25% required three responders among 21 patients in the first stage. Twenty-one patients who received a median of two earlier chemotherapy regimens were enrolled in the trial between March 2007 and May 2008. During the estradiol-only treatment phase, no patient had a PSA decline in excess of 50% and lesser PSA declines that ranged from 8.8 to 34.1% were seen in five patients. No patients achieved a > or =50% PSA decline following the addition of PPX and there were no responses in measurable disease. The median time to progression was 4 weeks. In conclusion, this regimen of low-dose transdermal estradiol induction followed by PPX does not have activity in taxane pretreated patients with castration-resistant prostate cancer. PMID- 20016366 TI - Sorafenib improves the survival of patients with advanced hepatocellular carcinoma: a meta-analysis of randomized trials. AB - There is no effective systemic therapy for patients with advanced hepatocellular carcinoma (HCC) except liver transplantation. Sorafenib, a multikinase inhibitor, has been shown to significantly increase overall survival (OS) in a randomized, placebo-controlled, phase III trial of patients with HCC (SHARP). The aim of this study was to evaluate the effectiveness of sorafenib for advanced HCC by carrying out a meta-analysis of randomized controlled trials that compared sorafenib-based therapy with other agent-based therapy. Randomized controlled trials comparing sorafenib or combined chemotherapy with placebo or combined chemotherapy in advanced HCC between 2000 and 2008 were identified and the data were extracted from reports. Outcomes analyzed were objective response rate, time to progression (TTP), OS, and toxicity. The summary hazard ratios (HRs), odds ratios, and their 95% confidence intervals (CIs) for mortality, objective response rate, and toxicity were estimated. All statistical tests were two-sided. Three trials including 924 patients were identified. Sorafenib-based chemotherapy was also associated with a 79% prolongation of TPP (HR = 0.58, 95% CI = 0.49-0.69, P<0.001), and a 37.3% increase in OS (HR = 0.66, 95% CI = 0.55-0.78, P<0.001). Despite significant increases in the frequencies of hand-foot syndrome and diarrhea in patients receiving sorafenib-containing chemotherapy, no significant difference in other toxic events was observed. This meta-analysis suggests that sorafenib-based chemotherapy is superior to placebo-based chemotherapy in terms of TPP and OS without increase in severe toxic effects. PMID- 20016367 TI - Aurora kinase inhibitors as anti-cancer therapy. AB - Aurora kinases are serine and threonine kinases that function as key regulators of the mitosis process. There are three distinct human aurora kinases known as Aurora A, Aurora B, and Aurora C. Aurora A and Aurora B are overexpressed in a number of human cancers including non-small cell lung cancer, glioblastomas, and upper gastrointestinal adenocarcinomas. Given their association with tumorigenesis, both Aurora A and Aurora B have been targeted for cancer therapy. Currently, a number of selective and nonselective aurora kinase inhibitors are being tested in preclinical and clinical settings as anti-tumor agents. We review the biology of human aurora kinases, followed by an overview of inhibitors undergoing current clinical investigations. PMID- 20016368 TI - Targeted therapy for nonsmall cell lung cancer: focusing on angiogenesis, the epidermal growth factor receptor and multikinase inhibitors. AB - Chemotherapy used to be the only available option to fight advanced nonsmall cell lung cancer. Platinum-based medication combined with taxanes, vinca alkaloids, and antimetabolites improved patient survival rates. Unfortunately, neoplasmatic diseases remain a global killer because chemotherapy benefits have reached a plateau and most patients are diagnosed at the metastatic stage. The urgent need for therapeutic agents, along with advances in the knowledge of the molecular events of oncogenesis, has resulted in the development of medication that specifically targets processes and pathways critical for tumor growth, such as angiogenesis and the epidermal growth factor receptor. Initially, inhibiting these pathways managed to prolong patient survival, although not to the extent desired. Moreover, targeted therapy combined with conventional cytotoxic agents has shown no superiority to chemotherapy alone in terms of patient survival. Hence, numerous multidynamic agents have appeared in the hope that they might help fight nonsmall cell lung cancer. However, no group of patients who will hopefully gain maximum benefit from such interventions has been clearly identified yet. This paper presents current evidence with regard to such novel agents and angiogenesis and epidermal growth factor inhibitors. PMID- 20016369 TI - Thymidine phosphorylase expression in metastatic sites is predictive for response in patients with colorectal cancer treated with continuous oral capecitabine and biweekly oxaliplatin. AB - The primary objective of this study was to determine the activity and safety profile of biweekly oxaliplatin combined with continuous oral capecitabine in the first-line treatment of metastatic colorectal cancer. A secondary endpoint was to investigate the correlation between thymidylate synthase and thymidine phosphorylase (TP) expression in metastatic tissues and tumor response. Forty-one patients received oral capecitabine 1331 mg/m every day combined with intravenous oxaliplatin 85 mg/m every 2 weeks. The overall response rate was 58.5% [95% confidence interval (CI): 43.3-73.6%], the median progression-free survival 9.4 months (95% CI: 7.7-11.2 months) and the median survival 22.3 months (95% CI: 16.1-27.5 months). There were no grade 4 toxicities, and grade 3 toxicity was also uncommon. High TP expression in metastatic tissue was significantly associated with response to treatment (P=0.019), and also with a trend towards a better median progression-free survival and overall survival compared with patients expressing low TP (P=0.056; P=0.073). This study suggests that biweekly oxaliplatin and continuous oral capecitabine is an active and well-tolerated chemotherapy regimen in the first-line treatment of metastatic colorectal cancer. Moreover, these findings add to a growing body of evidence that patients with high levels of intratumoral TP expression are the ideal candidates for capecitabine-based chemotherapy. PMID- 20016370 TI - Interleukin-2 treatment effect on imatinib pharmacokinetic, P-gp and BCRP expression in mice. AB - The aim of this study was to investigate the effect that recombinant interleukin 2 (rIL-2) (0.16 MUI/injection) had on the pharmacokinetics of imatinib (IM) in plasma. In this study, IM was given orally to mice at a dose of 150 mg/kg once a day for 11 days (from day 1 to 11) either alone or in combination with intraperitoneal injections of rIL-2 twice a day from day 8 to 11. Pharmacokinetic parameters were determined using WinNonLin software. Areas under the curve were compared using Bailer's method. The repeated administration of the rIL-2+IM combination was shown to have two pharmacokinetic advantages compared with repeated IM doses alone. In addition to the pharmacodynamic interest of this treatment, we found that the combined treatment significantly increased the IM Cmax (P<0.05) and significantly increased the IM trough concentration (C(24 h)) (P<0.01), which was always above the minimum therapeutic IM concentration (1 mumol/l) in plasma. Those pharmacokinetic modifications may be explained, in part, by a decrease in the P-glycoprotein expression in the three intestinal segments of the mice (duodenum, P<0.01; jejunum, P<0.05; and ileum, P<0.05) and a decrease in BCRP expression in the duodenum segment (P<0.05) due to rIL-2. In another experiment, we found a significant induction of intestinal P-glycoprotein expression in mice that had been given IM orally (150 mg/kg) twice a day for 11 days. It would be interesting to further investigate the IM disposition associated with rIL-2 treatment for clinical applications. PMID- 20016371 TI - Effective antitumor activity of paclitaxel-loaded poly (epsilon caprolactone)/pluronic F68 nanoparticles after intratumoral delivery into the murine breast cancer model. AB - A paclitaxel-loaded poly (epsilon]-caprolactone)(PCL)/pluronic F68 (F68) nanoparticle formulation was prepared as an intratumoral delivery system to assess its potential for future neoadjuvant chemotherapy application in the treatment of breast cancer. Paclitaxel-loaded nanoparticles were prepared by a solvent evaporation method using the self-synthesized PCL/F68 compound. Prepared nanoparticles were spherical with a rough, porous surface. As described in our earlier study, F68 was incorporated into the PCL matrix as both a pore-forming agent and to enhance drug release from the particles. A murine breast cancer model has shown that when using equivalent paclitaxel doses, paclitaxel-loaded PCL/F68 nanoparticles administered by a single intratumoral injection were more efficient in impeding tumor development than conventional paclitaxel injections administered by multiple intraperitoneal injections. In conclusion, paclitaxel loaded PCL/F68 nanoparticles can be delivered intratumorally and they effectively prevent tumor cell growth and establishment in a localized area. This treatment shows promise as a future neoadjuvant chemotherapy application in the treatment of breast cancer. PMID- 20016372 TI - Pulmonary toxicities of biologics: a review. AB - With the advancement of research in cancer treatment more and more drugs are being introduced for the treatment of cancer. In this review study, we have tried to look at some of the relatively newly introduced drugs, commonly referred to as biologics. The aim of this study was to review the very rare but fatal pulmonary toxicities (mostly interstitial lung disease) caused by these drugs. The drugs that were reviewed are rituximab, cetuximab, bevacizumab, alemtuzumab, and trastuzumab. This review basically aims at presenting a basic introduction (mechanism of action and indications of use) of these drugs followed by a summary of the incidence, various clinical presentations, diagnosis, treatment options, and outcome of patients around the world who presented with pulmonary toxicities caused by these drugs. PMID- 20016373 TI - Impact of early life stress on the reinforcing and behavioral-stimulant effects of psychostimulants in rhesus monkeys. AB - Early life stress has effects on behavior and stress reactivity, which are linked to enhanced sensitivity to stimulants in rodents. This study investigated whether rhesus monkeys that experienced early life stress would show altered sensitivity to the reinforcing effects of stimulants as compared with controls. Control (n=5) and maternally separated (n=4) monkeys were trained to self-administer cocaine (0.1 mg/kg/injection) under a second-order schedule of intravenous drug delivery. The rate of acquisition and subsequent dose-effect determinations for cocaine (0.01-1.0 mg/kg/injection) and amphetamine (0.003-0.3 mg/kg/injection) provided complementary measures of reinforcing effectiveness. In addition, stimulant induced increases in home cage activity and dopamine D2 receptor binding potential were quantified with positron emission tomography neuroimaging. Compared with controls, maternally separated monkeys showed lower responding during the acquisition of self-administration and in the dose-response curves for both stimulants, and significantly lower response rates during maintenance of cocaine self-administration. Maternally separated monkeys also failed to exhibit stimulant-induced increases in motor activity. Groups did not differ in dopamine D2 receptor binding potential in the caudate nucleus or the putamen. Taken together, the results of this study do not provide support for early life stress leading to enhanced vulnerability to stimulant use in the nonhuman primate model employed. PMID- 20016374 TI - Depressed expression of angiogenic growth factors in the subacute phase of myocardial ischemia: a mechanism behind the remodeling plateau? AB - BACKGROUND AND AIMS: To investigate whether, in the subacute phase of acute myocardial infarction, in the peri-infarcted area the expressions of the vascular endothelial growth factor (VEGF-A) and angiopoietin (Ang) ligand receptors are depressed, and whether overexpression of these angiogens counteracts a downregulation of myocardial function. METHODS: Acute myocardial infarction was induced by left anterior descending artery ligation and overexpression through injection of human VEGF-A165 and Ang-1 plasmids. The capillary and arteriolar densities, Akt-1 phosphorylation and citrate synthase activity were measured concurrent with the expression of VEGF-A, VEGFR1 and R2, Ang-1, Ang-2 and Tie-2. RESULTS: One day after AMI, VEGR-2 was unchanged but all other measured factors in the two families were upregulated. After day 2, the Ang-2 expression increased but other measured factors decreased. After gene transfer, the vascular supply, Akt phosphorylation and citrate synthase activity were higher in the peri infarcted area, where also the endogenous angiogenic growth factor expressions were increased. CONCLUSION: A rapid decrease in angiogenic stimulating factors occurs in the subacute phase of AMI and is related to a progressive decrease in myocardial contraction. A negative consequence of such a circuit is a successive reduction in the vascular supply and contractility in areas with reduced perfusion. These negative adaptations can be counteracted by angiogen overexpression. PMID- 20016375 TI - Increased O-linked beta-N-acetylglucosamine levels on proteins improves survival, reduces inflammation and organ damage 24 hours after trauma-hemorrhage in rats. AB - OBJECTIVE: To evaluate the effects of O-linked beta-N-acetylglucosamine (O GlcNAc) levels on survival, inflammation, and organ damage 24 hrs after trauma hemorrhage. We have previously shown that increasing protein O-GlcNAc levels by different mechanisms reduced inflammatory responses and improved organ function 2 hrs after trauma-hemorrhage. DESIGN: Prospective, randomized, controlled study. SETTING: Animal research laboratory. SUBJECTS: Male, adult Sprague-Dawley rats. INTERVENTIONS: Overnight fasted animals were subjected to either sham surgery or trauma-hemorrhage and during the resuscitation phase received glucosamine (270 mg/kg) to increase O-GlcNAc synthesis or O-(2-acetamido-2-deoxy-D glucopyranosylidene) amino N-phenyl carbamate (PUGNAc, 7 mg/kg) to inhibit O GlcNAc removal, or mannitol as control. MEASUREMENTS AND MAIN RESULTS: Survival was followed up for 24 hrs. Surviving rats were euthanized and inflammatory responses, and end organ injuries were assessed. Both glucosamine and PUGNAc increased 24-hr survival compared with controls (control: 53%, GN: 85%, PUGNAc: 86%, log-rank test, p < .05). PUGNAc attenuated the trauma-hemorrhage-induced increase in serum interleukin-6 (sham surgery: 8 +/- 6, control: 181 +/- 36, PUGNAc: 42 +/- 22 pg/mL, p < .05), alanine transaminase (sham surgery: 95 +/- 14, control: 297 +/- 56, PUGNAc: 126 +/- 21 IU, p < .05), aspartate transaminase (sham surgery: 536 +/- 110, control: 1661 +/- 215, PUGNAc: 897 +/- 155 IU, p < .05), and lactate dehydrogenase (sham surgery: 160 +/- 18, control: 1499 +/- 311, PUGNAc: 357 +/- 99 IU, p < .05); however, glucosamine had no effect on these serum parameters. Furthermore, PUGNAc but not glucosamine maintained O-GlcNAc levels in liver and lung and significantly attenuated the NF-kappaB DNA activation in the liver. In the liver and heart, increased inducible nitric oxide synthase expression was also attenuated in the PUGNAc-treated group. CONCLUSIONS: These results demonstrate that increasing O-GlcNAc with either glucosamine or PUGNAc improved 24-hr survival after trauma-hemorrhage. However, only PUGNAc treatment attenuated significantly the subsequent tissue injury and inflammatory responses, suggesting that inhibition of O-GlcNAc removal may represent a new therapeutic approach for the treatment of hypovolemic shock. PMID- 20016377 TI - Perceived barriers to therapeutic hypothermia for patients resuscitated from cardiac arrest: a qualitative study of emergency department and critical care workers. AB - OBJECTIVE: To identify the barriers to implementation of mild therapeutic hypothermia for adult survivors of cardiac arrest. Despite scientific evidence to support therapeutic hypothermia for resuscitated cardiac arrest patients, it is inconsistently and at times inadequately used. DESIGN: Qualitative study, using semistructured interviews. SETTING: A stratified random sample of 14 sites from an established network of 43 hospitals, including both community and tertiary care centers in Southern Ontario, Canada. PARTICIPANTS: Twenty-one intensive care unit and emergency department physicians and nurses. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: Purposive sampling was used to interview individuals who were most likely to be involved in the implementation and evaluation of the hypothermia protocol. All interviews were conducted by telephone by a clinician and a qualitative researcher. Interviews were recorded electronically and transcribed unless the participant declined to have the interview recorded. Untranscribed interviews were recorded as field notes and as direct quotations. New interviews were conducted until thematic saturation occurred. The analysis was completed through three phases of coding. Respondents identified lack of familiarity and availability of concrete therapeutic hypothermia protocols and process issues as the most frequent barriers. Process concerns included availability of equipment, equipment costs, and high workload demands for emergency nurses. Other barriers identified were variable nursing awareness, variable staff uptake, lack of agreement with supporting evidence, lack of interdisciplinary collaboration between the intensive care unit and emergency department, lack of interprofessional education between nurses and physicians, and challenges inherent in applying an intervention infrequently. CONCLUSIONS: This study demonstrated that the systematic adoption of a new intervention, therapeutic hypothermia, is met with interdependent generic, local, and individual barriers. A working awareness of the types of barriers that exist at multiple sites will assist in targeting specific knowledge translation strategies to improve adherence to evidence-based practice. PMID- 20016376 TI - Defining normal adrenal function testing in the intensive care unit setting: a canine study. AB - OBJECTIVE: To determine whether intensive care medicine therapies and testing influence hypothalamic-pituitary-adrenal test results. It is routine in intensive care medicine to measure hypothalamic-pituitary-adrenal function, commonly utilizing the adrenocorticotropic hormone stimulation test to diagnose absolute or relative adrenal insufficiency. DESIGN: Prospective, 96-hr animal study. SETTING: Research laboratory. SUBJECTS: Twenty-four healthy canines. INTERVENTIONS: Animals were randomized into two groups--awake and unrestrained or treated with intensive care medicine therapies, including sedation, intubation, and mechanical ventilation. Animals were further randomized to receive dexamethasone (or placebo) or undergo either a total of four or seven adrenocorticotropic hormone stimulation tests over 96 hrs. MEASUREMENTS AND MAIN RESULTS: Sedation, intubation, and mechanical ventilation transiently increased both basal and postadrenocorticotropic hormone total and free cortisol concentrations >2-fold as compared with baseline for the first 24 hrs (p < or = .05 for both). Performance of seven stimulation tests increased both basal and postadrenocorticotropic hormone total and free cortisol concentrations from baseline by >1.5-fold for the duration of the 96-hr study (p < or = .05). Neither sedation, intubation, and mechanical ventilation nor the performance of more stimulation tests affected delta cortisol measurements (total or free cortisol, p = NS). In contrast, dexamethasone suppressed basal total cortisol concentrations by >2-fold (p < or = .005) at all time points and transiently increased delta total cortisol by approximately 35% during the first 24 hrs of the study (p < or = .05). CONCLUSIONS: Total and free cortisol measurements--whether pre- or post- adrenocorticotropic hormone or as a calculated delta--were altered by intensive care therapies or frequent adrenocorticotropic hormone stimulation testing with one exception. Delta free cortisol was the only hypothalamic-pituitary-adrenal measurement unaffected by sedation, intubation, and mechanical ventilation, completion of more adrenocorticotropic hormone stimulation tests, or dexamethasone therapy. These findings support the need to determine normal ranges for hypothalamic-pituitary-adrenal testing in subjects receiving intensive care medicine before establishing laboratory criteria for the diagnosis of relative adrenal insufficiency. PMID- 20016378 TI - Anti-N-methyl-D-aspartate receptor antibodies: a potentially treatable cause of encephalitis in the intensive care unit. AB - OBJECTIVE: To report the occurrence of an unusual neurologic disorder requiring admission to the intensive care unit. DESIGN: Analysis of an observational cohort study of 31 patients with encephalitis admitted over a 4-yr period. SETTING: Neurologic intensive care unit in a tertiary referral center. PATIENTS: We identified N-methyl-D-aspartate receptor antibodies in six patients (two male and four female). All seropositive patients presented with a psychiatric prodrome, before developing seizures and obtundation requiring intensive care unit admission. They exhibited limb and truncal stereotypies and orofacial dyskinesias upon weaning sedation. Two patients had ovarian tumors. INTERVENTIONS: Patients were treated with sedation, antiepileptic drugs, and immunotherapy. One patient received a magnesium infusion and ketamine. MEASUREMENTS AND MAIN RESULTS: N methyl-D-aspartate receptor antibodies were identified in serum samples by an immunofluorescent cell-based assay. Three patients made a good but slow recovery; two were left with severe neurologic deficits; and one died after return to the referring hospital. These patients accounted for approximately 20% of all patients admitted with encephalitis to this referral center. CONCLUSIONS: N methyl-D-aspartate receptor antibodies should be tested in patients with hyperkinetic encephalitis and neuropsychiatric prodrome admitted to the intensive care unit. The disorder is probably not rare and is potentially treatable. PMID- 20016379 TI - Intensive care unit alarms--how many do we need? AB - OBJECTIVE: To validate cardiovascular alarms in critically ill patients in an experimental setting by generating a database of physiologic data and clinical alarm annotations, and report the current rate of alarms and their clinical validity. Currently, monitoring of physiologic parameters in critically ill patients is performed by alarm systems with high sensitivity, but low specificity. As a consequence, a multitude of alarms with potentially negative impact on the quality of care is generated. DESIGN: Prospective, observational, clinical study. SETTING: Medical intensive care unit of a university hospital. DATA SOURCE: Data from different medical intensive care unit patients were collected between January 2006 and May 2007. MEASUREMENTS AND MAIN RESULTS: Physiologic data at 1-sec intervals, monitor alarms, and alarm settings were extracted from the surveillance network. Video recordings were annotated with respect to alarm relevance and technical validity by an experienced physician. During 982 hrs of observation, 5934 alarms were annotated, corresponding to six alarms per hour. About 40% of all alarms did not correctly describe the patient condition and were classified as technically false; 68% of those were caused by manipulation. Only 885 (15%) of all alarms were considered clinically relevant. Most of the generated alarms were threshold alarms (70%) and were related to arterial blood pressure (45%). CONCLUSION: This study used a new approach of off line, video-based physician annotations, showing that even with modern monitoring systems most alarms are not clinically relevant. As the majority of alarms are simple threshold alarms, statistical methods may be suitable to help reduce the number of false-positive alarms. Our study is also intended to develop a reference database of annotated monitoring alarms for further application to alarm algorithm research. PMID- 20016380 TI - Passive leg raising is predictive of fluid responsiveness in spontaneously breathing patients with severe sepsis or acute pancreatitis. AB - OBJECTIVE: Rapid fluid loading is standard treatment for hypovolemia. Because volume expansion does not always improve hemodynamic status, predictive parameters of fluid responsiveness are needed. Passive leg raising is a reversible maneuver that mimics rapid volume expansion. Passive leg raising induced changes in stroke volume and its surrogates are reliable predictive indices of volume expansion responsiveness for mechanically ventilated patients. We hypothesized that the hemodynamic response to passive leg raising indicates fluid responsiveness in nonintubated patients without mechanical ventilation. DESIGN: Prospective study. SETTING: Intensive care unit of a general hospital. PATIENTS: We investigated consecutive nonintubated patients, without mechanical ventilation, considered for volume expansion. INTERVENTIONS: We assessed hemodynamic status at baseline, after passive leg raising, and after volume expansion (500 mL 6% hydroxyethyl starch infusion over 30 mins). MEASUREMENTS AND MAIN RESULTS: We measured stroke volume using transthoracic echocardiography, radial pulse pressure using an arterial catheter, and peak velocity of femoral artery flow using continuous Doppler. We calculated changes in stroke volume, pulse pressure, and velocity of femoral artery flow induced by passive leg raising (respectively, Deltastroke volume, Deltapulse pressure, and Deltavelocity of femoral artery flow). Among 34 patients included in this study, 14 had a stroke volume increase of >or=15% after volume expansion (responders). All patients included in the study had severe sepsis (n = 28; 82%) or acute pancreatitis (n = 6; 18%). The Deltastroke volume >or=10% predicted fluid responsiveness with sensitivity of 86% and specificity of 90%. The Deltapulse pressure >or=9% predicted fluid responsiveness with sensitivity of 79% and specificity of 85%. The Deltavelocity of femoral artery flow >or=8% predicted fluid responsiveness with sensitivity of 86% and specificity of 80%. CONCLUSIONS: Changes in stroke volume, radial pulse pressure, and peak velocity of femoral artery flow induced by passive leg raising are accurate and interchangeable indices for predicting fluid responsiveness in nonintubated patients with severe sepsis or acute pancreatitis. PMID- 20016381 TI - Impact of the administration of probiotics on the incidence of ventilator associated pneumonia: a meta-analysis of randomized controlled trials. AB - BACKGROUND: Previous reviews showed no benefit for the administration of probiotics in critically ill patients, but they did not focus on ventilator associated pneumonia. DESIGN: Meta-analysis of randomized controlled trials comparing probiotics and control in patients undergoing mechanical ventilation and reporting on incidence of ventilator-associated pneumonia. METHODS: PubMed, Scopus, Current Contents, Cochrane Central Register of Controlled Trials, and reference lists were searched. Weighted mean differences, pooled odds ratios, and 95% confidence intervals were calculated, implementing both the Mantel-Haenszel fixed effect and the DerSimonian-Laird random effects model. RESULTS: Five randomized controlled trials were included. Administration of probiotics, compared with control, was beneficial in terms of incidence of ventilator associated pneumonia (689 patients; fixed effect model: odds ratio, 0.61; 95% confidence interval, 0.41-0.91; random effects model: odds ratio, 0.55, 95% confidence interval, 0.31-0.98), length of intensive care unit stay (fixed effect model: weighted mean difference, -0.99 days; 95% confidence interval, -1.37- 0.61), and colonization of the respiratory tract with Pseudomonas aeruginosa (odds ratio, 0.35; 95% confidence interval, 0.13-0.93). However, no difference was revealed between comparators regarding intensive care unit mortality (odds ratio, 0.75; 95% confidence interval, 0.47-1.21), in-hospital mortality (odds ratio, 0.75; 95% confidence interval, 0.46-1.24), duration of mechanical ventilation (weighted mean difference, -0.01 days; 95% confidence interval, -0.31 -0.29), and diarrhea (odds ratio, 0.61; 95% confidence interval, 0.28-1.34). CONCLUSION: Administration of probiotics is associated with lower incidence of ventilator-associated pneumonia than control. Given the increasing antimicrobial resistance, this promising strategy deserves consideration in future studies, which should have active surveillance for probiotic-induced diseases. PMID- 20016382 TI - Caveolin-1 and Hsp70 interaction in microdissected proximal tubules from spontaneously hypertensive rats as an effect of Losartan. AB - BACKGROUND: Caveolin is required to traffic the AT1 receptor through the exocytic pathway. The chaperone Hsp70 regulates a diverse set of signaling pathways via their interactions with proteins. METHOD: Here we examined the AT1 receptor antagonist Losartan effect on caveolin-1 and Hsp70 protein association in spontaneously hypertensive rat (SHR) proximal tubules. Hsp70 involvement in Losartan oxidative stress regulation was also studied. Five-week-old SHRs were randomized for receiving Losartan (40 mg/kg per day) (SHRLos) or no treatment (SHRH2O) during 6 weeks. Wistar-Kyoto rats (WKY) were normotensive controls. RESULTS: By western blotting, the relative abundance of caveolin-1 was two-fold higher in microdissected proximal tubule membrane fractions from treated SHRs vs. WKYH2O. Hsp70 membrane translocation was demonstrated in SHRLos through out the up-regulation of Hsp70 expression in microdissected proximal tubule membrane fractions when compared with WKYH2O (P < 0.001). Conversely, decreased Hsp70 protein levels were shown in microdissected proximal tubule cytosol fraction from SHRLos (P < 0.01). Interaction between caveolin-1 and Hsp70 was further determined by coimmunoprecipitation and by immunofluorescence co-localization in SHRLos proximal tubule membranes. After membrane translocation of Hsp70, the decreased NADPH oxidase activity (RFU/microprot per min incubation) near controls demonstrated on microdissected proximal tubule membranes from SHRLos vs. SHRH2O (P < 0.01) was reversed by the preincubation with anti-Hsp70 antibody. In addition, interaction between Hsp70 and Nox4 was determined by the coimmunoprecipitation strategy showing that membrane overexpression of Hsp70 was associated with decreased Nox4 after Losartan treatment in SHRs. CONCLUSION: After Losartan administration interaction of caveolin-1 and Hsp70 was shown in SHR proximal tubules. Translocation of Hsp70 to proximal tubule membranes in SHRLos might exert a cytoprotective effect by down-regulation of NADPH subunits Nox4. PMID- 20016383 TI - Differential effects of alcohol intoxication on S100B levels following traumatic brain injury. AB - BACKGROUND: In an acute care setting, evaluation of traumatic brain injury (TBI) is often complicated by alcohol intoxication. The purpose of this study is to evaluate the clinical utility of the protein S100B as a biochemical marker for identifying brain injury in patients who are intoxicated at the time of injury. METHODS: The study participants were 160 patients who presented to a large urban Level I Trauma Centre in Vancouver, Canada. Patients were classified into four clinical groups (medical controls, trauma controls, mild TBI, and definite TBI) and two day-of-injury alcohol intoxication groups (i.e., sober and intoxicated). Blood samples were collected via venipuncture in heparinized tubes within 8 hours of injury. Measures of S100B concentration were obtained using a commercially available assay kit (Sangtec 100 Elisa). RESULTS: For those patients who were sober at the time of injury, higher S100B levels were associated with TBI when compared with other physical injuries and general medical complaints. However, for patients who were intoxicated at the time of injury, there were uniformly low S100B levels across all clinical groups. CONCLUSIONS: Although there seems to be a strong association between S100B levels and TBI, further research is required to establish the clinical role of S100B in patients with suspected TBI, particularly in patients whose clinical presentation is complicated by alcohol intoxication. PMID- 20016384 TI - Identifying targets for potential interventions to reduce rural trauma deaths: a population-based analysis. AB - BACKGROUND: Rural environments have consistently been characterized by high injury mortality rates. Although injury prevention efforts might be directed to reduce the frequency or severity of injury in rural environments, it is plausible that interventions directed to improve injury care in the rural settings might also play a significant role in reducing mortality. To test this hypothesis, we set out to examine the relationship between rurality and the setting in which patient death was most likely to occur. METHODS: This is a population-based retrospective cohort study evaluating all trauma deaths occurring in the province of Ontario, Canada, over the interval 2002 to 2003. Patient cohorts were defined by their potential to access trauma center care using two different approaches, rurality and timely access to trauma center care. RESULTS: There were 3,486 deaths over the study interval, yielding an overall injury mortality rate of 14.6 per 100,000 person-years. Overall, more than half of deaths occurred before reaching an emergency department (ED). Prehospital deaths were twice as likely in the most rural locations and in those with limited access to timely trauma center care. However, among patients surviving long enough to reach hospital, there was a threefold increase in the risk of ED death among those injured in a region with limited access to trauma center care. CONCLUSIONS: We demonstrate that a significant proportion of deaths occur in rural EDs. This study provides new insights into rural trauma deaths and suggests the potential value of targeted interventions at the policy and provider level to improve the delivery of preliminary trauma care in rural environments. PMID- 20016385 TI - Endoscopic retrograde cholangiopancreatography in patients with pancreatic trauma. AB - BACKGROUND: : Pancreatic injury occurs in from 3% to 12% of patients with abdominal trauma. In many instances, a lack of impressive findings in the first 24 hours leads to a delay in diagnosis. Because pancreatic duct disruption is the major cause of traumatic pancreatitis, we evaluated our experience with endoscopic retrograde cholangiopancreatography (ERCP) in patients suspected of having of having pancreatic injury. METHODS: : We reviewed the medical records of 26 patients evaluated perioperatively by ERCP for suspected pancreatic duct injury. The examinations were performed in the endoscopy suite or radiography special procedures or operating rooms under direct fluoroscopic control using fiberoptic or videooptic duodenoscopes. RESULTS: : Seventeen men and nine women with a mean age of 32.8 +/- 2.2 years suffered severe abdominal trauma. ERCP was performed in these patients a mean of 19 +/- 11.3 days after trauma. Seven patients underwent ERCP just before or at laparotomy. Eight of 26 (31%) patients were found to have intact pancreatic and bile ducts, whereas 18 (69%) patients had substantial findings unsuspected by pre-ERCP imaging. Nine of these 18 patients with documented ductal injury underwent endoscopic treatment alone without further surgical intervention, including pancreatic sphincterotomies and/or pancreatic ductal stenting. CONCLUSIONS: : ERCP is feasible and strongly indicated in the care of many patients with pancreatic trauma. Patient care and overall surgical and hospital needs may be substantially impacted by the use of both diagnostic and therapeutic endoscopic retrograde colongiopancreatography. PMID- 20016387 TI - Progressive displacement of scapula fractures. AB - BACKGROUND: Historically, minimally to moderately displaced extra-articular scapula fractures have been managed conservatively. It is possible that operative criteria could be met for a given patient if progressive displacement were to occur. We hypothesize that certain displaced scapula fractures progressively displace in early postinjury period. METHODS: A consecutive series of 49 patients with operatively managed extra-articular scapula fractures was evaluated to identify patients initially managed nonoperatively and later operatively because of progressive displacement. Three examiners then independently analyzed radiographs to measure the displacement between injury films and the films used to determine operative indications. A standard method for radiographic measurement of deformity was developed and was used to determine the degree of deformity at injury and at follow-up time when operative care was decided. RESULTS: Eight patients met inclusion criteria. All sustained a specific extra articular transverse fracture pattern Arbeitsgemeinschaft fur Osteosynthefragen/Orthopaedic Trauma Association (AO/OTA) Type A3, Ada-Miller Type IIC). Mean time interval between injury and repeat radiographs was 11.3 (range, 9-15) days. All patients displayed progressive displacement in at least one of the four measured parameters (medial/lateral displacement, angulation, translation, and glenopolar angle), according to the three independent examiners. Predominantly "good" inter- and intraobserver scores were achieved. CONCLUSIONS: Certain extra-articular scapula fractures may displace in postinjury period. We recommend close monitoring of conservatively treated fractures to screen for progressive displacement. PMID- 20016386 TI - Factors affecting the development of hypothalamus and pituitary lesions in fatal closed head injury: a prospective study. AB - OBJECTIVES: To determine the factors affecting the development of pituitary and hypothalamic lesions after fatal closed head injury. MATERIALS: Thirty-four patients with severe closed head injury succumbing to the effects of brain trauma before or during admission, whether managed conservatively or surgically, formed the study group. Clinical parameters, injury to death interval, radiologic data, and management details were taken into consideration. Autopsy was performed within 48 hours of death; hypothalamus and pituitary were carefully removed and evaluated for the presence of lesions on hematoxylin and eosin and immunohistochemical staining. RESULTS: Patients were categorized into early death group (n = 11, those succumbing before/on admission) and late death group (n = 23, those succumbing after admission). Histopathologic evaluation of pituitary revealed capsular hemorrhages in 50%, posterior pituitary hemorrhage in 25%, anterior pituitary infarct in 21.8%, and anterior pituitary hemorrhage in 6.2% patients. Hypothalamic hemorrhage was observed in 65.2% patients and infarcts in 17.3%. Lesions in hypothalamus and pituitary were significantly related to the presence of ventricular compression on computed tomography scan and survival of >24 hours after injury (p < 0.05). Capsular hemorrhage, anterior pituitary hemorrhage, and posterior pituitary hemorrhage were present in 40%, 10%, and 30% of the patients in the early death group when compared with 54.5%, 4.5%, and 22.7% of the patients in the late death group. Anterior pituitary infarcts were present in 10% of the patients with early deaths and 27.3% patients in the late death group. Hypothalamic hemorrhages were present in 44.4% of patients in early death and 78.6% in late death groups. Hypothalamic infarcts (40%) were present in the late death group only. Two patients (25%) in the early death group and 11 (84.6%) in the late death group had lesions in pituitary as well as hypothalamus (p < 0.05). CONCLUSIONS: Presence of ventricular compression on computed tomography scan and survival >24 hours after severe head injury has a significant correlation with the development of hypothalamic and pituitary lesions. Secondary insults account for a proportion of pituitary and hypothalamic lesions after trauma, which may be amenable to prevention by early intervention to treat raised intracranial pressure (ICP)/herniation. PMID- 20016388 TI - Prone or lateral? Use of the Kocher-Langenbeck approach to treat acetabular fractures. AB - PURPOSE: This retrospective, institutional review board approved study with no patients excluded was designed to test the hypothesis that the prone position is advantageous for repair of acetabular fractures, via a posterior approach. METHODS: In 104 consecutive cases fixed by open reduction and internal fixation using the Kocher-Langenbeck approach, 50 were performed in the prone position and 54 in the lateral position by four attending surgeons at a level I trauma center. These cases were assigned to each surgeon according to the call schedule and positioned on the operating table depending on the preference of the assigned surgeon. Arbeitsgemeinschaft fuer Osteosynthese/Association for the Study of Internal Fixation (AO/ASIF) classification and radiologic outcome were evaluated by standard radiographs of the hip and computerized tomography scans, demographic data were taken from the medical records. Except for the time from injury to surgery (p = 0.003), both groups were comparable, nevertheless the fractures were more severe in the prone one. RESULTS: With equivalent radiologic outcomes according to Matta, Brooker, Epstein, and Helfet between both groups, a significantly higher rate of infection (p = 0.017) and need for revision surgery (p = 0.009) were found in the prone group. CONCLUSION: No advantage to either position for the posterior approach to acetabular fractures could be found. Because most of the severe fractures were performed prone, we propose that the larger number of more difficult fractures in this group may cause an increased likelihood of loss of reduction. The higher infection rate in the prone group may be caused by the longer inpatient wait for definitive fixation, leading to a higher risk of nosocomial colonization. PMID- 20016389 TI - Frequent computed tomography scanning due to incomplete three-view X-ray imaging of the cervical spine. AB - BACKGROUND: Conventional C-spine imaging (3-view series) is still widely used in trauma patients, although the utilization of computed tomography (CT) scanning is increasing. The aim of this study was to analyze the value of conventional radiography and the frequency of subsequent CT scanning due to incompleteness of three-view series of the C-spine in adult blunt trauma patients. METHODS: We analyzed the data of a prospectively collected database including all patients between November 2005 and November 2007 treated in the trauma resuscitating room. We assessed the reasons for subsequent CT scanning after the three-view series according to the following classification: inevaluability, incompletion, evaluation of findings on three-view series or evaluation of unexplained, and persistent clinical symptoms. Furthermore, we evaluated possible predictors for incompleteness. RESULTS: Of 1,283 blunt trauma patients, 88 C-spine injuries were diagnosed with an overall incidence of 6.9%. One hundred fifty-nine patients (12%) had their C-spine cleared based on the NEXUS criteria and 12 died before C spine imaging could be performed. A total of 717 patients were primarily evaluated with three-view series and 395 patients primarily with CT scanning. Within the population with primarily three-view series, 249 (35%) were repeatedly incomplete and 16 (2%) were inevaluable. In the majority of the incomplete three view series, no apparent reason could be determined. However, the presence of clavicular fractures (resulting in incomplete radiographs in 68% vs. 34% without a fracture; p < 0.001) and rib fractures (56% vs. 34%; p = 0.008) were associated with incomplete three-view series. CONCLUSION: In more than one third of the patients primarily assessed with three-view X-ray series of the C-spine, the results are incomplete or inevaluable necessitating CT scanning. Although the majority of the incomplete series remain unexplained, we advise CT scanning in patients having clavicular and rib fractures because this increases the likelihood of obtaining incomplete three-view series. PMID- 20016390 TI - The value of sequential computed tomography scanning in anticoagulated patients suffering from minor head injury. AB - BACKGROUND: Since 1999, the Italian guidelines have been used at our department for the management of patients with mild head injury (MHI). According to these guidelines, a computed tomography (CT) scan should be obtained in all patients with coagulopathy and these should routinely undergo strict observation during the first 24 hours after injury; in addition they should have a control CT scan before discharge. With the increased use of anticoagulant therapy in the elderly population, admitting patients in such treatment with a MHI to the emergency rooms has become very common. The aim of our study was to evaluate the need of performing a control CT scan in patients on anticoagulation treatment who showed neither intracranial pathology on the first CT-scan nor neurologic worsening during the observation period. METHODS: We prospectively analyzed the course of all patients on anticoagulation treatment consecutively admitted to our unit between October 2005 and December 2006 who suffered from a MHI and showed a normal initial CT scan. All patients underwent strict observation during the first 24 hours after admission and had a control CT scan performed before discharge. RESULTS: One hundred thirty-seven patients were included in this study. Only two patients (1.4%) showed hemorrhagic changes. However, neither of them developed concomitant neurologic worsening nor needed admitting or surgery. CONCLUSION: According with our data, patients on anticoagulation treatment suffering from MHI could be managed with strict neurologic observation without routinely performing a control CT scan that can be reserved for the rare patients showing new clinical symptoms. PMID- 20016391 TI - A biomechanical comparison of a biodegradable volar locked plate with two titanium volar locked plates in a distal radius fracture model. AB - BACKGROUND: Volar plating is commonly used in the management of distal radius fractures; bioresorbable plates have attractive features. We compared a bioresorbable plate with a latest generation and an established locked titanium plate. METHODS: Twenty-four fresh-frozen radii (12 pairs) were assigned to three mean bone mineral density-matched groups of eight radii each. A standardized extraarticular distal radius fracture was created and plated using one implant type per group. Postplating stiffness and displacement were studied in a first axial-loading test (15 cycles at 250 N). Next, biodegradation was simulated by 4 weeks' immersion in phosphate-buffered saline, followed by a second axial test. Finally, the specimens underwent cyclic loading (2,400 cycles at 250 N). RESULTS: It is clear from the initial test that the LCP plate was significantly stiffer and displaced less than the bioresorbable plate. The outcome of the postimmersion tests is that one bioresorbable plate failed early on after 4 weeks' immersion, and the remaining bioresorbable plates and the T plates did not differ significantly. Cyclic tests conclude that the LCP plate was significantly superior to the other systems. One T plate and four of the bioresorbable plates failed, but none of the LCP plates failed. In the bioresorbable constructs, stability, time to failure, and bone mineral density were significantly correlated. CONCLUSIONS: The LCP plate was biomechanically superior and may be generally recommended for the volar plating of distal radius fractures. Except one plate failure, the bioresorbable plate was similar to the T plate in the quasi-static tests and should, therefore, be considered for clinical studies, with patient selection confined, initially, only to candidates with good bone stock quality. PMID- 20016392 TI - B16 melanoma secretomes and in vitro invasiveness: syntenin as an invasion modulator. AB - To characterize proteins involved in melanoma dissemination, protein profiles from B16F10 and B16Bl6 cells were compared, as only B16Bl6 cells give pulmonary metastases after subcutaneous graft. As B16F10 and B16Bl6 cells had the same invasive capacities in vitro, we wondered whether their extracellular content could be different and correlate with their metastatic properties. We have shown that B16F10 and B16Bl6 culture cell supernatants have different modulatory effects on HT1080 fibrosarcoma cell invasion in Matrigel-coated chambers. B16Bl6 supernatants significantly enhanced HT1080 in vitro invasion as compared with B16F10 ones, suggesting differences in their protein profiles. Indeed, proteomic analysis allowed the identification of 18 differential proteins. Among the proteins with a higher concentration in B16Bl6 supernanants, lactate dehydrogenase B, M2 pyruvate kinase, cathepsin D, and galectin 1 were involved in the melanoma aggressiveness signature. Interestingly, several Gag retroviral proteins, as well as syntenin, were found mainly in the B16F10 secretome. Although its intracellular form is known as an aggressive melanoma marker, we show for the first time that syntenin was actively secreted and could reduce the invasion process, probably by protein interactions in the B16 model. PMID- 20016393 TI - Early visual-evoked potential acuity and future behavioral acuity in cortical visual impairment. AB - PURPOSE: Cortical Visual Impairment (CVI) is bilateral visual impairment caused by damage to the posterior visual pathway. Both preferential looking and sweep visual-evoked potential (VEP) can be used to measure visual acuity. The purpose of this study was to determine if an early VEP measure of acuity is related to a young patient's future behavioral acuity. METHODS: The visual acuity of 33 patients with CVI was assessed using the sweep VEP and a behavioral measure on two occasions. The median age of the patients at the initial visit was 4.8 years (range: 1.3 to 19.2 years), and they were followed for an average of 6.9 years (SD: 3.5 years). RESULTS: The mean initial VEP acuity was 20/135 (0.735 logMAR), and the mean initial behavioral acuity was 20/475 (1.242 logMAR). The average difference between the two initial measures of acuity was 0.55 log unit, with the behavioral measure reporting a poorer visual acuity in all patients. However, the mean final behavioral acuity was 20/150 (0.741 logMAR), and the average difference between the initial VEP acuity and the final behavioral acuity was only 0.01 log unit. Therefore, the initial VEP measure was not statistically different from the final behavioral measure (t = 0.11; dF = 32; p = 0.45). CONCLUSIONS: Even though the initial VEP measure was much better than the initial behavioral measure, the initial VEP measure was similar to the behavioral visual acuity measured approximately 7 years later. Sweep VEP testing can be used as a predictive tool for at least the lower limit of future behavioral acuity in young patients with CVI. PMID- 20016395 TI - Infectious complications of peripherally inserted central venous catheters in children. AB - BACKGROUND: Peripherally inserted central venous catheters (PICCs) are frequently used in infants and children. However, only limited data are available on current infectious and noninfectious complications of PICCs in this patient population. The aim of the study is to define the current rate of infectious and noninfectious complications of PICCs, the causative agents, and to define possible risk factors associated with these complications. METHODS: A prospective surveillance study was conducted at the Schneider Children Medical Center of Israel. All patients in whom a PICC was inserted between August 2004 and October 2006 were included. Demographic, clinical, and microbiologic data were collected by a questionnaire completed by the anesthesiologist at the time of insertion and removal of each catheter. Items covered epidemiologic, clinical, and microbiologic data. RESULTS: A total of 279 PICCs were inserted in 221 patients. There were no complications at insertion. Mean dwell time was 30 days. One hundred seventy-seven (63%) of all PICC placements were free of complications. Twenty-six catheters (9.3%) were dislodged accidentally; 38 (13.6%) were removed for mechanical problems: tears in 5, leaks in 12, and obstructions in 21; 38 (13.6%) were removed for an infectious complication: phlebitis in 13 (4.6%, 1.5/1000 PICC days), exit-site infection in 10 (3.5%, 1.1/1000 PICC days), PICC associated bloodstream infection in 12 (4.3%, 1.4/1000 PICC days), and PICC related bloodstream infection in 4 (1.4%, 0.4/1000 PICC days); 15 more were removed for presumed infection (5.3%, 1.7/1000 PICC days). On multivariate analysis, composite indication for PICC use and older patient age were significantly associated with infectious complications. CONCLUSIONS: PICCs are safe and may be used for prolonged periods. In our center, the rates of infectious complications are lower than for tunneled central venous catheters. Accidental dislodgement is not uncommon and may be prevented by use of sutures, occlusive dressing, and education of patients, families, and medical staff. PMID- 20016394 TI - Characteristics of children with Shigella encephalopathy: experience from a large urban diarrhea treatment center in Bangladesh. AB - BACKGROUND: Encephalopathy is a serious neurologic complication from shigellosis often with fatal outcome but data on simple clinical and sociodemographic predicting factors are limited. We sought to identify those factors in children. METHODS: In this unmatched case-control design, children of both sexes, aged 0 to 15 years having positive stool culture for Shigella from 1997 to 2006 were studied. Children with Shigella encephalopathy constituted the cases (n = 29), whereas randomly selected children with shigellosis having no encephalopathy constituted controls (n = 87). RESULTS: The case-fatality was significantly higher among the cases than among controls (7% vs. 0%, RR: 4.2, CI: 3.0-5.9, P = 0.013). In logistic regression analysis, children with Shigella encephalopathy more often had an illiterate father (OR: 5.4, 95% CI: 1.1-27.1, P = 0.038), stopped breast-feeding in the neonatal period (OR: 41.3, 95% CI: 4.2-403.1, P = 0.001), had dehydrating diarrhea (OR: 9.9, 95% CI: 2.1-45.9, P = 0.004) with a duration of <1 day (OR: 29.4, 95% CI: 2.1-43.2, P = 0.004), and were more likely to be severely stunted (OR: 6.4, 95% CI: 1.2-34.9, P = 0.033). CONCLUSION: Education of parents about the value of exclusive breast-feeding and of prompt hydration in children with Shigella is critical to minimize morbidity and deaths. PMID- 20016396 TI - Nonvertical, nonsexual transmission of human immunodeficiency virus in children. AB - In a cohort of 450 human immunodeficiency virus (HIV)-infected children followed up in Maputo, Mozambique, 22 were born to HIV-uninfected mothers and had no history of sexual abuse. A case record review of these nonvertically, nonsexually infected children as well as a case-control study strongly suggests health care as possible source of infection for many of these children. This facet of the pediatric HIV epidemic should not be overlooked. PMID- 20016397 TI - Population based external validation of a European predictive model for respiratory syncytial virus hospitalization of premature infants born 33 to 35 weeks of gestational age. AB - Prospectively collected population-based data on 2529 Danish infants born at 33 to 35 weeks of gestation were used to validate an European predictive model of respiratory syncytial virus (RSV) hospitalization. The model was found to be robust with a diagnostic accuracy of 65.9% to distinguish between RSV hospitalized versus non-RSV-hospitalized Danish infants born at 33 to 35 weeks of gestation. PMID- 20016398 TI - Heterozygous variants of multidrug and toxin extrusions (MATE1 and MATE2-K) have little influence on the disposition of metformin in diabetic patients. AB - Multidrug and toxin extrusions (MATE1/SLC47A1 and MATE2-K/SLC47A2) play important roles in the renal excretion of metformin. We have previously identified the nonsynonymous MATE variants with functional defects at low allelic frequencies. The purpose of this study was to evaluate the effects of heterozygous MATE variants on the disposition of metformin in mice and humans. Pharmacokinetic parameters of metformin in Mate1(+ or -) heterozygous mice were comparable with those in Mate1(+ or +) wild-type mice. Among 48 Japanese diabetic patients, seven patients carried heterozygous MATE variant and no patient carried homozygous MATE variant. There was no significant difference in oral clearance of metformin with or without heterozygous MATE variants. In addition, creatinine clearance, but not heterozygous MATE variants, significantly improved the model fit of metformin clearance by statistical analysis using the nonlinear mixed-effects modeling program. In conclusion, heterozygous MATE variants could not influence the disposition of metformin in diabetic patients. PMID- 20016399 TI - No association between regulator of G-protein signaling 9 (RGS9) and schizophrenia in a Jewish population. PMID- 20016400 TI - Lack of genetic association of the PLA2G4C locus with schizophrenia in case control samples. PMID- 20016401 TI - Association between major depressive disorder and a functional polymorphism of the 5-hydroxytryptamine (serotonin) transporter gene: a meta-analysis. AB - OBJECTIVES: A functional polymorphism in the promoter region of the 5 hydroxytryptamine (serotonin) transporter (5-HTT) gene, termed 5-HTTLPR, alters transcription of the 5-HTT gene. The short variation (S allele) produces less transcriptional efficiency of serotonin, which can partly account for psychiatric disorders. Despite strong biological plausibility, the relationship between 5 HTTLPR and the risk of major depressive disorder (MDD) is unclear. To elucidate the relationship, we applied meta-analysis techniques to molecular studies of 5 HTTLPR and MDD. METHODS: A total of 22 articles were identified from MEDLINE through March 2008, using the search keywords 'depression,' '5-HTTLPR', and 'polymorphism.' The authors assessed the evidence of genotypic association using STATA Version 8.2. RESULTS: Summary frequencies of the S allele of 5-HTTLPR among Caucasians and Asians based on the random effects model were 42.1% [95% confidence interval (CI) = 40.5-43.6] and 76.8% (95% CI = 73.9-79.7), respectively. The distribution of the S allele was significantly different between Asians and Caucasians (P<0.001). The SS genotype was significantly associated with an increased risk of MDD among Caucasian populations (odds ratio = 1.41, 95% CI = 1.15-1.72), although there was no significant association among Asians. CONCLUSION: Although the summary risk for developing MDD in individuals with the 'at-risk' SS genotype of 5-HTTLPR may be small, MDD is such a common disease that even a small increase in risk translates to a large number of excess MDD cases in the population. Thus, 5-HTT may be a candidate MDD susceptibility gene. PMID- 20016402 TI - Adenosine-mediated attenuation of the innate immune response for only those who need it? The tailored potential of pentoxifylline. PMID- 20016403 TI - Proteomic analysis of post-hemorrhagic shock mesenteric lymph. AB - Recent studies have documented the association of mesenteric lymphatic route with adult respiratory distress syndrome and multiple organ failure after hemorrhagic shock. However, the mediators and mechanisms of the toxic effects of mesenteric lymph remain unclear. This study aimed to identify mediators or biomarkers in the mesenteric lymph through comparative proteomic analysis. Fourteen mature male Sprague-Dawley rats were randomly divided and subjected to trauma (laparotomy) plus hemorrhagic shock or trauma plus sham shock. Mesenteric lymph samples were collected before shock and at 3 h after resuscitation from hemorrhagic shock (or sham shock). To investigate changes in proteome profiles between preshock and 3-h postshock (or 3-h post-sham shock) mesenteric lymph samples, two-dimensional gel electrophoresis and matrix-assisted laser desorption ionization time-of-flight mass spectrometry were performed. We found a more than 2-fold change in abundance of 31 protein spots in the lymph samples. Mass spectrometry analyses identified 12 distinct proteins. Four proteins were consistently upregulated in the 3-h postshock lymph samples, including serum albumin precursor, two isoforms of cytoplasmic actin, complement C3 precursor, and major urinary protein precursor. Two proteins, including haptoglobin and one unidentified protein, were consistently downregulated. The deregulation of these proteins was confirmed by Western blots. Most of these altered proteins are functionally implicated in tissue inflammation. The findings of this study provide a starting point for investigating the functions of these proteins in hemorrhagic shock-induced lung injury and hold great promise for the development of potential therapeutic interventions. PMID- 20016404 TI - Hyperoxia suppresses excessive superoxide anion radical generation in blood, oxidative stress, early inflammation, and endothelial injury in forebrain ischemia/reperfusion rats: laboratory study. AB - This study used an electrochemical O2. sensor to investigate the effects of hyperoxia on generation of the superoxide radical (O2.) in the jugular vein during forebrain I/R in rats. Twenty-eight male Wistar rats were allocated to a sham group (n = 7; sham-treated rats with inspired oxygen fraction [FiO2] of 0.4), a hemorrhagic shock and reperfusion (HS/R) group (n = 7; HS without carotid artery occlusion and reperfusion with FiO2 of 0.4), a normoxia group (n = 7; forebrain ischemia produced by bilateral carotid arteries occlusion with HS and reperfusion with FiO2 of 0.4), and a hyperoxia group (n = 7; forebrain ischemia with FiO2 of 0.4 and reperfusion with FiO2 of 1.0). The jugular venous O2. current was measured for 10 min during forebrain ischemia and for 120 min after reperfusion. The O2. current increased gradually during forebrain ischemia in the three groups other than the sham group. Immediately after reperfusion, the current showed a marked increase in the normoxia group and a pronounced decrease in the hyperoxia group. Levels of brain and plasma malondialdehyde, high-mobility group box 1 protein, and intercellular adhesion molecule 1 were significantly attenuated in the hyperoxia group relative to those in the normoxia group. In conclusion, hyperoxia suppressed jugular venous O2. generation and malondialdehyde, high-mobility group box 1, and intercellular adhesion molecule 1 in the brain and plasma in the acute phase of cerebral I/R. Thus, the administration of 100% oxygen immediately after reperfusion suppresses oxidative stress and early inflammation in cerebral I/R. PMID- 20016405 TI - Early increase in arterial lactate concentration under epinephrine infusion is associated with a better prognosis during shock. AB - To determine whether an epinephrine-induced early increase in arterial lactate concentration can prognosticate the outcome during shock state, we conducted a retrospective study in a 16-bed medical intensive care unit of a teaching hospital in France. One hundred consecutive patients admitted because of a shock state irrespective of etiology and treated with epinephrine were included. Patients were not enrolled if they received epinephrine administration before intensive care unit admission. Sequential arterial lactate measurements were performed at the time of epinephrine infusion (H0) and 4 h later (H4) in which Deltalactate was defined as (100 x [arterial lactate(H4)-arterial lactate(H0)]/arterial lactate(H0)) and expressed as a percentage. Etiology of shock was septic (82%), cardiogenic (10%), or hemorrhagic (8%). Twenty-eight-day mortality rate was 72%. At admission, arterial lactate concentration was elevated (4.96 +/- 3.8 mmol/L) and was further increased upon epinephrine administration, reaching a peak at H4 (8.22 +/- 3.66). When patients were stratified according to their outcome, nonsurvivors displayed the same pattern as survivors, although with a significant upward shift in values (ANOVA, P = 0.0003). The Sequential Organ Failure Assessment score and Deltalactate were the only variables associated with the 28-day risk of death, with an odds ratio of 1.32 (95% confidence interval [CI], 1.06-1.65; P = 0.01) and 0.99 (95% CI, 0.99-0.99; P = 0.03), respectively, in multivariate analysis. At a value of 100%, Deltalactate predicted death, with a 71% sensitivity (95% CI, 51%-87%) and a 67% specificity (95% CI, 43%-85%). Kaplan-Meier survival analysis confirmed this finding, with a 52.4% death rate among patients with Deltalactate greater than 100 comparatively to 84.7% when Deltalactate was less than 100 (log-rank test, P = 0.0002). An adapted response (lactate production) to a pharmacological trigger (epinephrine) is associated with better prognosis during shock of various etiologies. PMID- 20016407 TI - Polymorphisms in PARP, IL1B, IL4, IL10, C1INH, DEFB1, and DEFA4 in meningococcal disease in three populations. AB - The pathogenesis of meningococcal infections involves activation of the complement system, proinflammatory and anti-inflammatory mediators, antimicrobial peptides, and apoptosis. We hypothesized that variations in genes encoding these products are involved in the susceptibility to and severity of pediatric meningococcal infections. Polymorphisms in poly (adenosine diphosphate-ribose) polymerase (PARP), serine protease C1 inhibitor (C1INH), IL4, IL10 and IL1B, alpha-defensin 4, and beta-defensin 1 (DEFB1) were analyzed in two independent Caucasian case control cohorts from the United Kingdom and the Netherlands and in a family-based transmission disequilibrium test cohort from the UK. In the UK case control cohort, the DEFB1 -44 G/G homozygous genotype was overrepresented in patients with meningococcal disease compared with the G/C and C/C genotypes when combined (odds ratio, 1.57; 95% confidence interval, 1.12-2.20). The transmission disequilibrium test analysis did not confirm this, but did find an association and linkage of the IL4 -524 and the C1INH 480 polymorphisms with susceptibility to meningococcal infection. Hematological failure was present more often in UK patients with the DEFB1 -44 G/G genotype compared with the C allele carriers (odds ratio, 2.17; 95% confidence interval, 1.22-3.85). Additional studies are necessary to elucidate the conflicting results obtained for the DEFB1, IL4, and C1INH polymorphisms and their role in susceptibility to and severity of meningococcal disease. PMID- 20016408 TI - Chronic inflammatory diseases in children are more severe in girls. AB - In humans and animal models, females express higher immune reactivity and more robust inflammatory responses. We analyzed the expression of current inflammatory markers in 149 children (74 girls and 75 boys) with three chronic inflammatory diseases: 50 with asthma, 47 with cystic fibrosis, and 52 with sickle cell anemia to evaluate the potential differences in clinical response according to sex. Data including temperature, neutrophil count (NC), and C-reactive protein were recorded for each patient at several time points according to his/her disease. In asthma, NC was higher in girls than in males (P < 0.02), as were doses of cortisone (P < 0.04) or inhaled bronchodilators (P < 0.01) received at recovery. In cystic fibrosis, NC became significantly higher in girls at age 5 years (P < 0.003), whereas episodes of infection and antibiotic administration were already significantly more frequent in girls at age 2 years (P < 0.02 and P < 0.05, respectively). In sickle cell anemia, the number of crises since diagnosis and number of acute chest syndrome episodes were significantly higher in girls (P < 0.01 and P < 0.05, respectively). Our study extends the documentation of a relationship between sex, inflammatory markers, and clinical outcome in prepubescent children, suggesting a genetic predetermination is more likely than hormonal influence. PMID- 20016409 TI - Role of natural killer dendritic cells in host resistance against Pseudomonas aeruginosa infection after thermal injury in mice. AB - The contributions of dendritic cells (DCs) and natural killer dendritic cells (NKDCs) on host antibacterial innate immunities have been described. We have previously reported that mice with partial-thickness burn injuries (PT-burn mice) are resistant to burn wound infections, whereas mice with full-thickness burn injuries (FT-burn mice) are susceptible. In this study, the effect of burn stress on the appearance and properties of DCs and NKDCs was investigated in two different murine models of thermal injury. Dendritic cells isolated from PT-burn mice produced CCL3 and IL-12, whereas these soluble factors were not produced by DCs from FT-burn mice. As compared with unburned mouse controls, a large number of NKDCs were isolated from the DC preparations from PT-burn mice, whereas fewer NKDCs were detected in the DC preparations from FT-burn mice. Nonobese diabetic severe combined immunodeficiency mice inoculated with NKDCs were shown to be resistant against a lethal s.c. Pseudomonas aeruginosa infection. These results strongly suggest that NKDCs influenced by partial-thickness burn injury play a role on the resistance of PT-burn mice to P. aeruginosa s.c. infection. PMID- 20016410 TI - Llama heavy-chain antibody fragments efficiently remove toxic shock syndrome toxin 1 from plasma in vitro but not in experimental porcine septic shock. AB - Staphylococcus aureus produces the superantigen toxic shock syndrome toxin 1 (TSST-1). When the bacterium invades the human circulation, this toxin can induce life-threatening gram-positive sepsis. Current sepsis treatment does not remove bacterial toxins. Variable domains of llama heavy-chain antibodies (VHH) against toxic shock syndrome toxin 1 ([alpha]-TSST-1 VHH) were previously found to be effective in vitro. We hypothesized that removing TSST-1 with [alpha]-TSST-1 VHH hemofiltration filters would ameliorate experimental sepsis in pigs. After assessing in vitro whether timely removing TSST-1 interrupted TSST-1-induced mononuclear cell TNF-[alpha] production, VHH-coated filters were applied in a porcine sepsis model. Clinical course, survival, plasma interferon [gamma], and TSST-1 levels were similar with and without VHH-coated filters as were TSST-1 concentrations before and after the VHH filter. Plasma TSST-1 levels were much lower than anticipated from the distribution of the amount of infused TSST-1, suggesting compartmentalization to space or adhesion to surface not accessible to hemofiltration or pheresis techniques. Removing TSST-1 from plasma was feasible in vitro. However, the [alpha]-TSST-1 VHH adsorption filter-based technique was ineffective in vivo, indicating that improvement of VHH-based hemofiltration is required. Sequestration likely prevented the adequate removal of TSST-1. The latter warrants further investigation of TSST-1 distribution and clearance in vivo. PMID- 20016412 TI - Endotracheal suctioning is basic intensive care or is it?: commentary on article by Copnell et al. on page 405. PMID- 20016413 TI - Anesthesia-induced neurodegeneration in fetal rat brains. AB - We investigated the extent of isoflurane-induced neurodegeneration on the fetuses of pregnant rats exposed in utero. Pregnant rats at gestational d 21 were divided into three experimental groups. Rats in the control group spontaneously breathed 100% oxygen for 1 h. Rats in the treatment groups breathed either 1.3 or 3% isoflurane in 100% oxygen through an endotracheal tube, with mechanical ventilation for 1 h. Rat pups were delivered by cesarian section 6 h after treatment, and fetal blood was sampled from the left ventricle of each fetal heart and evaluated for S100beta. Fetal brains were then evaluated for apoptosis, using caspase-3 immunohistochemistry in the CA1 region of the hippocampus and the retrosplenial cortex (RS). The 3% isoflurane treatment group showed significantly higher levels of S100beta levels and significantly increased average densities of total caspase-3-positive cells in the CA1 hippocampus and RS cortex compared with the control and the 1.3% isoflurane groups. There were no differences in S100beta levels or densities of caspase-3-positive cells between the control and 1.3% isoflurane groups. Isoflurane at a concentration of 3% for 1 h increased neurodegeneration in the hippocampal CA1 area and the retrosplenial cortex in the developing brain of fetal rats. PMID- 20016415 TI - Pandemic Influenza A (H1N1). PMID- 20016417 TI - Letter to editor on "Myocardial inflammation, cellular death, and viral detection in sudden infant death caused by SIDS, suffocation, or myocarditis". PMID- 20016418 TI - New direction for the Southern Medical Journal. PMID- 20016419 TI - The metabolic syndrome: underdiagnosed and undertreated. PMID- 20016420 TI - Mechanical ventilation simplified. PMID- 20016421 TI - Rectal stents as an alternative to surgery. PMID- 20016422 TI - Alternative approaches to pancreatic duct injury. PMID- 20016423 TI - Comparative role of angiotensin receptor blockers versus other agents in the management of hypertension, cardiovascular disease and nephropathy. PMID- 20016425 TI - Mass casualties from acute inhalation of chlorine gas. AB - OBJECTIVES: Chlorine gas is a potent pulmonary irritant that affects the mucous membranes and induces severe disturbances of pulmonary gas exchange within minutes of inhalation. The present study evaluated an extraordinary type of mass inhalational exposure. MATERIAL AND METHODS: Clinical reports of 25 soldiers who were admitted to the emergency department of Maresal Cakmak Military Hospital, Erzurum were retrospectively evaluated. All patients were exposed to chlorine gas as a result of mixing sodium hypochlorite with hydrochloric acid during cleaning activities. RESULTS: All patients were male and the mean age of patients was 22.04+/-2.98 years. The main symptoms were coughing and dyspnea in 18 patients (72%). Forced expiratory volume in 1 second (FEV1) and FEV1/forced volume capacity (FVC) ratio were found to be normal in all patients but FVC and peak expiratory flow (PEF) were below the normal range (80%) in 9 patients (36%). All patients received warmed humidified oxygen combined with nebulized salbutamol. Inhaled budesonide and nebulized sodium bicarbonate were ordered additionally for 19 patients (76%). Thirteen patients (52%) were discharged from the emergency department and 12 patients (48%) were hospitalized. No mortality was observed. CONCLUSION: Chlorine gas is a potent pulmonary irritant that causes acute damage in both the upper and lower respiratory tract. We suggest that inhaled steroids combined with nebulized sodium bicarbonate could be a safe and effective alternative for the treatment of symptomatic patients. Education of the public about the dangers of mixing of hypochlorite bleach with acidic cleaning agents is also very important. PMID- 20016424 TI - The association between Chlamydia pneumoniae and metabolic syndrome in Taiwanese adults. AB - OBJECTIVE: Both Chlamydia pneumoniae (Cp) seropositivity and metabolic syndrome (MeS) have been identified as risk factors for atherosclerosis and cardiovascular disease (CVD), but the association between Cp seropositivity and MeS has not been extensively studied. BACKGROUND: We recruited participants from Taiwanese adults who received an annual health examination at a medical facility. We measured anti Cp IgG antibodies (Cp-IgG) by enzyme-linked immunosorbent assay and defined Cp IgG seropositivity using 20 RU/mL as the cutoff point. RESULTS: In the 3633 study participants, the overall Cp-IgG seropositive rate was 45.5%. Male gender, older age, white collar jobs, smoking, diabetes mellitus, and hypertension were associated with a higher Cp-IgG seropositive rate (P<0.05). Participants with MeS had a higher Cp-IgG seropositive rate (62.8% vs. 37.2%, P<0.001). Multiple logistic regression revealed that positive Cp-IgG (odds ratio [OR]=1.4, 95% confidence interval [CI]: 1.1-1.9), male gender (OR=5.0, 95% CI: 3.3-7.6) and older age(OR=2.6, 95% CI: 1.1-6.1 for age>or=40 years old) were independent predictors for MeS. CONCLUSION: There is a positive association between Cp seropositivity and MeS, which supports the role of infection in the pathogenesis of atherosclerosis and CVD. PMID- 20016426 TI - Epiploic appendagitis: the great mimicker. AB - Epiploic appendagitis is a benign, self-limited, clinicopathological, and radiological entity that has become more commonly recognized. The clinical presentation resembles a variety of other medical and surgical intra-abdominal conditions and may easily be confused with many of them. Computed tomography (CT) demonstrates characteristic features that help make the diagnosis in most cases, thus avoiding unneeded treatment or surgery. Here, we discuss a case series of five patients who presented with abdominal pain and were ultimately diagnosed with epiploic appendagitis. PMID- 20016427 TI - Prayers in the clinic: how pediatric physicians respond. AB - BACKGROUND: Physicians and researchers have recently paid increased attention to prayer in physician-patient interactions. Research focuses more on attitudinal questions about whether physicians and/or patients think prayer is relevant than on actual data about when and how prayer comes up in the clinic and how physicians respond. We focus on pediatric physicians to investigate: 1) how prayer enters clinical contexts and 2) how physicians respond. METHODS: We examined in-depth interviews with 30 academic pediatricians and pediatric oncologists. All of these physicians were employed by the most highly ranked hospitals according to US News and World Report. RESULTS: In close to 100% of cases when the subject of prayer came up in clinical contexts, it was patients and families who raised it. Patients and families mostly talked about prayer in response to a seriously ill or dying child. When it was raised, pediatric physicians responded to prayer by participating; accommodating but not participating; reframing; and directing families to other resources. CONCLUSIONS: Physicians wanted to respect patients and families around the topic of prayer. They negotiated between patient/family requests, the specific situation, and their own comfort levels to respond in one of four ways. Their four responses allowed researchers to generate hypotheses about the independent variables that influence how pediatric physicians respond to prayer. Asking how prayer actually came up in clinical situations rather than how patients and/or physicians thought it should be raised, better informs ongoing conversations about the significance of prayer in physician-patient interactions. PMID- 20016428 TI - High levels of autoantibodies against catalase and superoxide dismutase in nasopharyngeal carcinoma. AB - INTRODUCTION: Autoantibodies against the antioxidant enzymes have been described in Epstein-Barr virus-associated diseases. Here, we hypothesize that Epstein-Barr virus (EBV), which is associated with nasopharyngeal carcinoma (NPC), induces anticatalase and/or antisuperoxide dismutase autoantibodies that inhibit catalase and/or superoxide dismutase activities and thereby contribute to the oxidative stress status described in this pathology. METHODS: Using a standard enzyme linked immunosorbent assay (ELISA), the levels of immunoglobulin G (IgG), and M (IgM) directed against catalase and superoxide dismutase in the sera of 30 NPC patients and 30 healthy control individuals were evaluated. The antioxidative profile was tested among the same patients by measuring serum catalase and superoxide dismutase activities. To investigate the implication of EBV in the establishment of autoantibody production in NPC patients, a correlation study between serological testing for EBV viral capsid antigen immunoglobulin G (VCA IgG) and autoantibodies against both enzymes was undertaken. FINDINGS: The levels of IgG against superoxide dismutase and catalase were found to be increased in sera patients compared to controls (P<0.001). NPC patients exhibited decreased catalase (P<0.001) and superoxide dismutase activities (P<0.001) in their sera. However, a positive correlation between superoxide dismutase IgM antibody and IgG antibody titers to VCA (P<0.05, r=0.483, n=21) was found. A positive correlation between catalase (IgM) antibodies and IgG antibody titers to VCA (P<0.05, r=0.546, n=30) was also found. CONCLUSION: High levels of anticatalase and antisuperoxide dismutase antibodies in the sera of NPC patients were found. PMID- 20016430 TI - Bronchus-associated lymphoid tissue lymphomas. AB - BACKGROUND: : Bronchus-associated Lymphoid Tissue (BALT) lymphomas are a rare type of extranodal marginal zone lymphomas. They comprise 1% of lymphomas and more than two-thirds of all primary non-Hodgkin lymphoma (NHL) of the lung. BALT lymphomas arise from the bronchus-associated lymphoid tissue. METHODS: This report describes five cases of BALT lymphoma and discusses the pathogenesis, diagnosis, prognosis and treatment of BALT lymphomas. RESULTS: In our cohort of patients, patients were managed with surgery, watchful waiting, chemotherapy, immunotherapy, and chemoimmunotherapy. The outcomes are excellent and projected 5 year survival is 100%. DISCUSSION: BALT lymphomas are associated with chronic inflammation, and they are often asymptomatic. They have an indolent course and the survival outcome is excellent with different treatment modalities such as surgery, watchful waiting, radiotherapy, chemotherapy, immunotherapy or chemoimmunotherapy. PMID- 20016431 TI - Surgical margins in breast conservation therapy: how much should we excise? AB - BACKGROUND: Breast conservation therapy (BCT) has become the standard of treatment for early stage breast cancer, and the surgical margin was one of the important factors that affected risk of local recurrence. This review looks at the safe margin for BCT in early stage invasive breast cancer and ductal carcinoma in situ (DCIS). METHODS: Published literature abstracted in Medline was searched using the gateway site from the US National Library of Medicine. CONCLUSIONS: A positive margin is associated with increased risk of local recurrence after BCT for invasive breast cancer and DCIS. However there was no cut off for the margin width and the significance of a close margin remains controversial. It was generally accepted that the risk of local recurrence was low if the margin was >or=10 mm while margins that were <2 mm were considered inadequate. The surgeon needs to balance the risk between local recurrence and cosmesis in planning BCT so that the prognosis is not compromised. PMID- 20016432 TI - Basic invasive mechanical ventilation. AB - Invasive mechanical ventilation is a lifesaving intervention for patients with respiratory failure. The most commonly used modes of mechanical ventilation are assist-control, synchronized intermittent mandatory ventilation, and pressure support ventilation. When employed as a diagnostic tool, the ventilator provides data on the static compliance of the respiratory system and airway resistance. The clinical scenario and the data obtained from the ventilator allow the clinician to provide effective and safe invasive mechanical ventilation through manipulation of the ventilator settings. While life-sustaining in many circumstances, mechanical ventilation may also be toxic and should be withdrawn when clinically appropriate. PMID- 20016433 TI - Narcolepsy: clinical decision making for the primary care physician. AB - Narcolepsy is a serious neurological condition in which patients are overcome by persistent, excessive feelings of fatigue and drowsiness. In addition to chronic fatigue, patients with narcolepsy often succumb to intermittent, uncontrollable periods where they abruptly fall asleep during waking hours. In addition to episodic bouts of daytime sleeping, narcoleptics also exhibit cataplexy, sleep paralysis, and hypnagogic and hypnopompic hallucinations. Unfortunately, many individuals with narcolepsy remain undiagnosed and therefore, untreated, posing a risk to themselves and those around them. There is currently no cure for this lifelong disease. Nonetheless, narcolepsy can be effectively managed with medications, lifestyle changes, and the peripheral support of individuals such as family members, coworkers, and other casual relations. PMID- 20016434 TI - Pancreatic injury. AB - Injury to the pancreas, because of its retroperitoneal location, is a rare occurrence, most commonly seen with penetrating injuries (gun shot or stab wounds). Blunt trauma to the pancreas accounts for only 25% of the cases. Pancreatic injuries are associated with high morbidity and mortality due to accompanying vascular and duodenal injuries. Pancreatic injuries are not always easy to diagnose resulting in life threatening complications. Physical examination as well as serum amylase is not diagnostic following blunt trauma. Computed tomography (CT) scan can delineate the injury or transaction of the pancreas. Endoscopic retrograde pancreaticography (ERCP) is the main diagnostic modality for evaluation of the main pancreatic duct. Unrecognized ductal injury leads to pancreatic pseudocyst, fistula, abscess, and other complications. Management depends upon the severity of the pancreatic injury as well as associated injuries. Damage control surgery in hemodynamic unstable patients reduces morbidity and mortality. PMID- 20016435 TI - Palliation of malignant rectal obstruction from invasive prostate cancer with multiple overlapping self-expanding metal stents. AB - Self-expandable metal stents (SEMS) are used for colonic neoplastic and extracolonic metastatic obstruction relief. Limited data exists on their use for locally invasive prostate cancer. We describe a unique approach using overlapping SEMS to alleviate a rectosigmoid obstruction from locally invasive prostate cancer. A patient with locally advanced prostate cancer presented with obstipation and lymphedema. Placement of overlapping rectosigmoid SEMS was performed, relieving the visualized rectosigmoid obstruction. PMID- 20016436 TI - Prolonged respiratory illness after single overnight continuous positive airway pressure humidification: endotoxin as the suspect. AB - A patient developed a prolonged respiratory illness after a single overnight use of tap water to humidify air supplied by a constant positive airway pressure (CPAP) device, which she had previously used for six years without difficulty. During those years, she used only distilled water for this purpose, as instructed by her sleep specialist. Analysis of the well water supplying her home showed no microorganisms, metals or other analytes likely to have caused her illness, but endotoxin was found at concentrations well above that recommended by the U.S. Pharmacopeia, as a maximum in water which may be inhaled as an aerosol. PMID- 20016437 TI - Brugada syndrome unmasked by lithium. AB - A 38-year-old man was brought by emergency medical service after resuscitation following cardiac arrest. The patient was found pulseless with a wide complex tachycardia. The patient had bipolar disorder and was on lithium, lamotrigine, and ziprasidone. His electrolytes and lithium levels were normal. An electrocardiogram (EKG) was performed the next day and showed type 1 Brugada pattern. Lithium was held. Electrophysiologists made a diagnosis of drug-unmasked Brugada syndrome. Lithium can unmask Brugada syndrome through its ability to block sodium channels, even at subtherapeutic concentrations. Physicians need to be aware of this potentially fatal drug effect and should monitor EKGs of patients on lithium. PMID- 20016438 TI - Priapism and risperidone. AB - A patient taking oral risperidone while using cocaine and alcohol presented with priapism shortly after long-acting, injectable risperidone was prescribed. Another case of priapism was diagnosed in a sickle cell disease patient who was on oral risperidone. A penectomy due to necrosis was required in the first case, while the other patient regained erectile function. Physicians should be aware of priapism-inducing/predisposing factors, such as sickle cell disease, certain medicines or drugs of abuse, and antipsychotic medications. Risperidone is an established cause of priapism. It occurs especially in males with sickle cell disease; use of antipsychotic drugs and/or cocaine further increases risk. PMID- 20016439 TI - Nonvenereal sclerosing lymphangitis of the penis. AB - A 30-year-old man presented to the genitourinary medicine clinic with the sudden appearance of an asymptomatic, 2 mm wide, flesh-colored, firm, and cord-like lesion encircling the coronal sulcus circumferentially following vigorous sexual intercourse with his wife. An initial diagnosis of sexually transmitted disease associated penile swelling was entertained. He was given antibiotics for this condition. Failure of resolution and negative screening tests for sexually transmitted diseases prompted a skin biopsy and extensive literature search, with an ultimate diagnosis of nonvenereal sclerosing lymphangitis of the penis, a rare, self-limiting condition. Reassurance and abstinence of sexual intercourse for a few weeks led to complete recovery. PMID- 20016440 TI - Primary biliary cirrhosis in an elderly man with normal liver tests. AB - Primary biliary cirrhosis (PBC) is a cholestatic liver disease that usually presents in middle-aged women. Women are about ten times more commonly affected than men. The case of a 78-year-old man with PBC who presented with pruritus is discussed. Liver biopsy and antimitochondrial antibody serology were diagnostic of early-stage PBC. PMID- 20016441 TI - Pericarditis revealing a relapse of sarcoidosis. PMID- 20016442 TI - Regarding "The third wave: medical tourism in the 21st century". PMID- 20016443 TI - Dilated cardiomyopathy associated with Bechterew disease and myopathy. PMID- 20016444 TI - Cervical spinal epidural abscess complicated with cerebral salt wasting. PMID- 20016445 TI - Heyde syndrome: a common but less recognized complex of aortic stenosis and bleeding intestinal angiodysplasia. PMID- 20016446 TI - Metastases with hypoadrenalism: diagnostic parallels and diagnostic traps. PMID- 20016447 TI - Multiple faces of sarcoidosis mimicking an underlying disease: don't forget immunoglobulin G. PMID- 20016449 TI - Developments of magnetic resonance imaging and spectroscopy in the field of molecular imaging. PMID- 20016450 TI - Magnetic resonance imaging and spectroscopy methods for molecular imaging. AB - Magnetic resonance imaging (MRI), one of the most powerful imaging modalities available for clinical diagnosis, has contributed significantly to phenotyping of transgenic organisms and to cellular imaging and is now gaining importance in the field of molecular imaging. Its advantage is the ability to provide in vivo information with high resolution and good soft tissue contrast as compared to established other molecular imaging methods. MRI can non-invasively report on cell localisation and migration with detailed anatomical background information, which is of great interest in cellular therapies. Recent technological advances and contrast generation strategies aim to bring MRI beyond cellular imaging to the detection of functional changes in vivo. MR based monitoring of molecular processes, requires the development of contrast agents and targeting methods as well as improvements in the methods sensitivity. Here, an overview is provided on advanced MR technologies and contrast generation strategies for this purpose. This includes MRI and MR spectroscopic methods for molecular imaging and various approaches for targeted and responsive contrast generation to visualize functional changes of particular cells. A description of different methods is provided, as well as the potentials and challenges of MR techniques for the visualization of molecular processes in vivo. PMID- 20016451 TI - Lanthanide complexes for magnetic resonance and optical molecular imaging. AB - Lanthanide complexes are more and more used in biomedical imaging as contrast agents (CA). The development of these paramagnetic complexes as CA for medical magnetic resonance imaging (MRI) and luminescent probes for optical imaging is very complementary. Gd complexes are well known as CA for MRI and Eu/Tb complexes are often used in microscopy or fluorescence imaging. Each imaging technique has its limitation: low sensitivity but high spatial resolution for MRI and limited penetration but high sensitivity for optical imaging. A bimodal agent can be used for these two methods and give more informations, they can be visualized simultaneously by light and MR imaging. Such compounds are based on the coordination chemistry of the lanthanide ions with an organic ligand to form a stable complex and on the properties of the lanthanide ions. Gd complexes with a chromophore allows also the luminescent detection. This review describes the properties of the lanthanide ions and of their complexes and gives some typical applications of the complexes. The luminescence properties show high quantum yield and long luminescence lifetimes. The relaxometric data of the Gd complexes are comparable or higher than commercial and clinically Gd-DTPA derivatives. PMID- 20016452 TI - Hyperpolarized agents for advanced MRI investigations. AB - The strong signal enhancement attainable by hyperpolarization methods has allowed the detection of heteronuclei in magnetic resonance imaging (MRI), allowing to obtain high quality images with very high signal to noise ratios in few seconds. The four methods to produce hyperpolarized molecules, i.e. the "brute force" approach, optical pumping of noble gases, parahydrogen induced polarization (PHIP) and dynamic nuclear polarization (DNP), are reported. The applications of hyperpolarized probes to MRI range from vascular imaging to interventional imaging and perfusion studies, up to the emerging and challenging field of molecular/metabolic imaging. In fact, the high signal intensities achievable by using hyperpolarized molecules make it possible to detect and image the metabolic products formed upon the administration of the hyperpolarized agent. The most striking examples are surveyed, including the use of hyperpolarized 13C-pyruvate in tumor diagnosis and stadiation, and in myocardium perfusion and activity studies, as well as the recently reported proposal of using 13C-bicarbonate as agent for pH-mapping in vivo. PMID- 20016453 TI - Proton magnetic resonance spectroscopy imaging in the study of human brain cancer. AB - Magnetic resonance spectroscopic imaging (MRSI) is a non-invasive imaging technique that provides metabolic information on brain tumor. This biochemical information can be processed and presented as density maps of several metabolites, among them N-acetylaspartate (marker of neuronal viability), choline (marker of membrane turnover), creatine (related to the energy state of the cells), myo-Inositol (exclusively found in astrocytes), lipids and lactate (observed in necrosis and other pathological processes) which mean relevant information in the context of brain tumors. Thus, this technique is a multiparametrical molecular imaging method that can complete the magnetic resonance imaging (MRI) study enabling the detection of biochemical patterns of different features and aspects of brain tumors. In this article, the role of MRSI as a molecular imaging technique to provide biochemical information on human brain tumors is reviewed. The most frequent questions and situations in the study of human brain tumors in clinical settings will be considered, as well as the distinction of neoplastic lesions from non neoplastic, the tumor type identification, the study of heterogeneity and infiltration of normal appearing white matter and the therapy following with detection of side effects. The great amount of data in MRSI acquisition compared to the single voxel techniques requires the use of automated methods of quantification, but the possibility to obtain self-reference in the non-affected areas allows different strategies for data handling and interpretation, as presented in the literature. The combination of MRSI with other physiological MRI techniques and positron emission tomography is also included in this review. PMID- 20016454 TI - 13C NMR tracers in neurochemistry: implications for molecular imaging. AB - An overview of 13C nuclear magnetic resonance (NMR) spectroscopy methods and their applications in the study of the metabolism of brain cells in vitro and in the in vivo brain is presented as well as their implications for modern molecular imaging techniques. Various topics will be discussed, such as general properties of the 13C NMR spectrum, 13C NMR spectroscopy acquisition protocols, determination of fractional 13C enrichment, 13C(2H) NMR methodologies, and the use of 13C hyperpolarized substrates for NMR spectroscopy and imaging. Some illustrative applications are described, both in vitro and in vivo. PMID- 20016455 TI - Differences in hippocampal metabolism between amnestic and non-amnestic MCI subjects: automated FDG-PET image analysis. AB - AIM: The aim of this study was to assess whether 18F-fluorodeoxyglucose positron emission tomography differentiates amnestic (aMCI) from single-non-amnestic mild cognitive impairment (snaMCI) with executive dysfunction. METHODS: Sixteen aMCI subjects (62% females, age 75+/-8 years) and 14 snaMCI subjects (71% females, age 74+/-6 years) underwent [18F]FDG-PET and clinical follow-up. Comparisons between MCI subgroups and with seven cognitively normal elderly subjects were performed using SPM2. RESULTS: At baseline aMCI and snaMCI exhibited a similar pattern of hypometabolism, mostly in the posterior cingulate gyrus, as compared with controls. In the comparison between the MCI subtypes, the aMCI subjects showed reduced metabolism in the medial temporal lobes (MTL) (hippocampus, fusiform gyrus and amygdala). At follow-up 12 aMCI developed Alzheimer's disease (AD), while snaMCI had a heterogeneous course, including five subjects who developed Lewy body dementia. CONCLUSIONS: The patterns of altered brain metabolism in aMCI and snaMCI subjects compared to controls are similar and do not provide evidence for making clinical distinctions between them. Comparison between the two MCI subtypes showed MTL hypometabolism in aMCI subjects, possibly reflecting the fact that most had prodromal AD. PMID- 20016456 TI - A practical dead time correction method in planar activity quantification for dosimetry during radionuclide therapy. AB - AIM: Gamma camera saturation is the first quantification problem in dosimetric studies following therapeutic administrations of 131I labeled radiopharmaceuticals. A new approach for dead time correction (DTC) is here proposed. It employs planar whole-body (WB) images without the need of standard radionuclide sources or of preliminary phantom calibrations. METHODS: Step and shoot WB acquisitions of the patient are required. A program was developed to compensate for the image discontinuities ("Continuity DTC method") between two adjacent static fields of view (FOVs) caused by different dead time count losses. For its validation, authors used two 99mTc 6 GBq phantom scans after administration of six patients with 131I labeled agents with different statistics and ten clinical scans taken between 16 h and 48 h after administration of 131I labeled agents, whose activity ranged from 4 to 10 GBq. The deviation from true decay corrected counts on phantoms and the constancy of monitor point-source counts in different patients' FOVs (root mean square error and maximum deviation) served as figures of merit. The accuracy of absorbed dose calculation was also estimated by comparison with the standard source correction method, computing the area under the time activity curve (AUC) of six lesions. RESULTS: With respect to the true phantom counts, corrected images gave excellent results, giving a 6% maximum deviation. For what concerns the other figures of merit, continuity DTC reduced the average root mean square error from 36% to 2% and the mean maximum deviation from 50% to 2%, on phantom, while from 51% to 32/28% (absence/presence of triple energy window scatter correction) and from 72% to 21/14% on patients. Mean compensation of AUC gave a correction of +56% with our method, while +78% with standard source method. CONCLUSIONS: The "Continuity DTC method" is a useful tool in dosimetry during nuclear medicine treatment, showing good accuracy. Moreover, since it does not require the use of any source, it provides with several advantages in terms of practicability and applicability, with respect to the standard source method and to methods based on the count rate characteristic curve. PMID- 20016457 TI - Decreased [99mTc]Sestamibi uptake with dobutamine versus dipyridamole stress. AB - AIM: Animal studies suggest an interference of dobutamine on [99mTc]sestamibi uptake. In this study dobutamine was compared to dipyridamole rest-stress [99mTc]sestamibi uptake ratio (UR). METHODS: Twenty-five patients with suspect coronary artery disease (CAD) underwent rest, dobutamine, and dipyridamole [99mTc]sestamibi SPECT at 24-h intervals and coronary angiography. UR was calculated separately for each coronary territory considering injected dose and acquisition delay. RESULTS: There were 38 CAD territories in 20 patients. On a patient basis, dipyridamole SPECT sensitivity was 85%, versus 70% for dobutamine. On a territory basis, sensitivity was 66% versus 42% (P<0.05), and specificity 92% versus 86%, respectively for dipyridamole versus dobutamine. In the 38 CAD territories, dipyridamole UR was -4.1+/-29.4%, and dobutamine UR was -13.1+/ 19.9% (P<0.05). In the 37 no-CAD territories, UR was 34+/-23.6% for dipyridamole and -0.4+/-17.8% for dobutamine (P<0.0001). UR difference between CAD versus no CAD territories was larger using dipyridamole (P<0.0001) than dobutamine (P<0.005). CONCLUSIONS: The UR comparison confirms that [99mTc]sestamibi uptake underestimates the blood flow heterogeneity induced by dobutamine more than that produced by dipyridamole. PMID- 20016459 TI - Extraction of the EPP component from the surface EMG. AB - A surface electromyogram (EMG), especially when recorded near the neuromuscular junction, is expected to contain the endplate potential (EPP) component which can be extracted with an appropriate signal filter. Two factors are important: the EMG must be recorded in monopolar fashion, and the recording must be done so the low frequency signal corresponding the EPP is not eliminated. This report explains how to extract the EPP component from the EMG of the masseter muscle in a human subject. The surface EMG is recorded from eight sites using traditional disc electrodes aligned along over the muscle, with equal inter-electrode distance from the zygomatic arch to the angle of mandible in response to quick gum clenching. A reference electrode is placed on the tip of the nose. The EPP component is extracted from the raw EMGs by applying a high-cut digital filter (2nd dimension Butterworth filter) with a range of 10-35 Hz. When the filter is set to 10 Hz, the extracted EPP wave deflects either negative or positive depending on the recording site. The difference in the polarity reflects the sink source relation of the end plate current, with the site showing the most negative deflection corresponding to the neuromuscular junction. In the case of the masseter muscle, the neuromuscular junction is estimated to be located in the inferior portion close to the angle of mandible. The EPP component exhibits an interesting oscillation when the cut-off frequency of the high-cut digital filter is set to 30 Hz. The EPP oscillation indicates that muscle contraction is adjusted in an intermittent manner. Abnormal tremors accompanying various sorts of diseases may be substantially due to this EPP oscillation, which becomes slower and is difficult to cease. PMID- 20016461 TI - A super-agonist of growth hormone-releasing hormone causes rapid improvement of nutritional status in patients with chronic kidney disease. AB - Chronic kidney disease is frequently associated with protein-energy wasting related to chronic inflammation and a resistance to anabolic hormones such as insulin and growth hormone (GH). In this study, we determined whether a new GH releasing hormone super-agonist (AKL-0707) improved the anabolism and nutritional status of nondialyzed patients with stage 4-5 chronic kidney disease randomized to twice daily injections of the super-agonist or placebo. After 28 days, this treatment significantly increased 24-h GH secretion by almost 400%, without altering the frequency or rhythmicity of secretory bursts or fractional pulsatile GH release, and doubled the serum insulin-like growth factor-1 level. There was a significant change in the Subjective Global Assessment from 'mildly to moderately malnourished' to 'well-nourished' in 6 of 9 patients receiving AKL-0707 but in none of 10 placebo-treated patients. By dual-energy X-ray absorptiometry, both the mean fat-free mass and the body mineral content increased, but fat mass decreased, all significantly. In the AKL-0707-treated group, both serum urea and normalized protein equivalent of nitrogen appearance significantly decreased with no change in dietary protein intake, indicating a protein anabolic effect of treatment. Thus, our study shows that stimulation of endogenous GH secretion by AKL-0707 overcomes uremic catabolism of patients with advanced chronic kidney disease. PMID- 20016462 TI - Novel aspects of complement in kidney injury. AB - Complement activation is integral to the development and progression of multiple forms of kidney disease. The liver is the principal source of serum complement, but various kidney cell types and bone marrow-derived immune cells can produce a full array of complement proteins. Locally produced and activated complement yields cleavage products that function as vital intermediaries, amplifying inflammation in ischemia-reperfusion injury and transplant rejection, among other pathological states. Additional new studies indicate that during cognate T-cell antigen presenting cell interactions, both cell types produce alternative pathway complement components. The resultant activation products have an essential role in T-cell activation, expansion, and differentiation, which in turn has a profound impact on the development of immune-mediated kidney disease. The recognition of an expanded role for kidney cell-derived and immune cell-produced complement as pathogenic to the kidney supports the need for future studies to test the efficacy of complement inhibitors in the prevention and/or treatment of selected kidney diseases. PMID- 20016463 TI - Mutations in components of complement influence the outcome of Factor I associated atypical hemolytic uremic syndrome. AB - Genetic studies have shown that mutations of complement inhibitors such as membrane cofactor protein, Factors H, I, or B and C3 predispose patients to atypical hemolytic uremic syndrome (aHUS). Factor I is a circulating serine protease that inhibits complement by degrading C3b and up to now only a few mutations in the CFI gene have been characterized. In a large cohort of 202 patients with aHUS, we identified 23 patients carrying exonic mutations in CFI. Their overall clinical outcome was unfavorable, as half died or developed end stage renal disease after their first syndrome episode. Eight patients with CFI mutations carried at least one additional known genetic risk factor for aHUS, such as a mutation in MCP, CFH, C3 or CFB; a compound heterozygous second mutation in CFI; or mutations in both the MCP and CFH genes. Five patients exhibited homozygous deletion of the Factor H-related protein 1 (CFHR-1) gene. Ten patients with aHUS had one mutation in their CFI gene (Factor I-aHUS), resulting in a quantitative or functional Factor I deficiency. Patients with a complete deletion of the CFHR-1 gene had a significantly higher risk of a bad prognosis compared with those with one Factor I mutation as their unique vulnerability feature. Our results emphasize the necessity of genetic screening for all susceptibility factors in patients with aHUS. PMID- 20016464 TI - Atorvastatin attenuates murine anti-glomerular basement membrane glomerulonephritis. AB - Statins mediate many of their protective effects by lowering lipids as well as by modulating inflammation. Here, we studied their potential immunomodulatory role in renal inflammation using an autoimmune mouse model of anti-glomerular basement membrane glomerulonephritis. Oral treatment with Atorvastatin dramatically reduced albuminuria and histological changes in the kidneys as compared to vehicle-treated control animals. There was a significant decrease in the Th1 and Th17 response in the regional lymph nodes draining the kidneys. This systemic effect was accompanied by decreased infiltration of the kidneys with inflammatory CD4(+) T and Th17 cells, macrophages, and neutrophils in statin-treated mice. Regulatory T cells were not altered in their number, FoxP3 expression, or suppressive capacity, but their interleukin-10 production was significantly increased by statin treatment. Hence, Atorvastatin systemically and locally decreased the Th1 and Th17 response, thereby protecting the mice against anti glomerular basement membrane glomerulonephritis. Whether statins can be used to treat human autoimmune renal diseases will require more direct studies. PMID- 20016465 TI - Verapamil inhibits calcification and matrix vesicle activity of bovine vascular smooth muscle cells. AB - Calcium channel activity in vascular smooth muscle cells is a critical component during vascular calcification and formation of matrix vesicles. Here, we examined whether the blockade of L-type calcium channels inhibits these functions. Bovine vascular smooth muscle cells or rat aorta organ cultures were incubated in media known to promote calcification and treated with the L-type calcium channel inhibitors verapamil, nifedipine, or nimodipine. The phenylalkylamine, verapamil, significantly decreased calcification of the vascular smooth muscle cells and rat aorta, in a dose-dependent manner, whereas the dihydropyridines, nifedipine and nimodipine, had no effect. Furthermore, verapamil, but not nifedipine, significantly decreased the alkaline phosphatase activity of bovine vascular smooth muscle cells. Verapamil pretreatment of the cells also inhibited matrix vesicle alkaline phosphatase activity and reduced the ability of these matrix vesicles to subsequently calcify on a type I collagen extracellular matrix scaffold. As L-type channels are blocked by verapamil and dihydropyridines, we suggest that verapamil inhibits vascular smooth muscle mineralization and matrix vesicle activity by mechanisms other than the simple blockade of this calcium channel activity. PMID- 20016466 TI - Genotype-phenotype correlation in primary hyperoxaluria type 1: the p.Gly170Arg AGXT mutation is associated with a better outcome. AB - We sought to ascertain the long-term outcome and genotype-phenotype correlations available for primary hyperoxaluria type 1 in a large retrospective cohort study. We examined the clinical history of 155 patients (129 families primarily from Western Europe, North Africa, or the Middle East) as well as the enzymatic or genetic diagnosis. The median age at first symptom was 4 years, and at diagnosis 7.7 years, at which time 43% had reached end-stage renal disease. Presentations included: (1) early nephrocalcinosis and infantile renal failure, (2) recurrent urolithiasis and progressive renal failure diagnosed during childhood, (3) late onset with occasional stone passage diagnosed in adulthood, (4) diagnosis occurring on post-transplantation recurrence, and (5) family screening. The cumulative patient survival was 95, 86, and 74% at ages 10, 30, and 50 years, respectively, with the cumulative renal survival of 81, 59, 41, and 10% at ages 10, 20, 30, and 50 years, respectively; 72 patients had undergone a total of 97 transplantations. Among the 136 patients with DNA analysis, the most common mutation was p.Gly170Arg (allelic frequency 21.5%), with a median age at end stage renal disease of 47 years for homozygotes, 35 years for heterozygotes, and 21 years for other mutations. Our results underscore the severe prognosis of primary hyperoxaluria type 1 and the necessity for early diagnosis and treatment, as well as confirm a better prognosis of the p.Gly170Arg mutation. PMID- 20016467 TI - Blood pressure in chronic kidney disease stage 5D-report from a Kidney Disease: Improving Global Outcomes controversies conference. AB - Management of blood pressure (BP) in patients with chronic kidney disease receiving dialysis (stage 5D) provides a significant challenge for healthcare professionals. The association between BP and cardiovascular disease risk has been well studied in the general population; however, in dialysis patients, physiological and dialysis-related mechanisms influencing BP are complex, and the associated risk is poorly understood. In stage 5D, BP is determined by the complex interplay of fluid volume and prescription of post-dialysis target weight, sodium load, the renin-angiotensin and sympathetic nervous systems, and diverse exogenous factors, such as administration of erythropoiesis-stimulating agents, the type and timing of administration of antihypertensive drugs, and dialysate composition. Management of BP in this population requires both generally applicable plans and individualization in order to determine the BP target and the treatment regimen. This report summarizes the deliberations and recommendations of a conference sponsored by the Kidney Disease: Improving Global Outcomes (KDIGO) to address the following questions: (1) what is the optimal BP treatment target in relation to end-organ damage and outcomes in dialysis patients; (2) how should antihypertensive drugs be used in dialysis patients; and (3) what nonpharmacological therapies can be considered in achieving BP targets? The conference report will augment the KDIGO clinical practice guideline on blood pressure in chronic kidney disease stages 1-5, which is currently under development. PMID- 20016468 TI - Parathyroid cell resistance to fibroblast growth factor 23 in secondary hyperparathyroidism of chronic kidney disease. AB - Although fibroblast growth factor 23 (FGF23) acting through its receptor Klotho FGFR1c decreases parathyroid hormone expression, this hormone is increased in chronic kidney disease despite an elevated serum FGF23. We measured possible factors that might contribute to the resistance of parathyroid glands to FGF23 in rats with the dietary adenine-induced model of chronic kidney disease. Quantitative immunohistochemical and reverse transcription-PCR analysis using laser capture microscopy showed that both Klotho and FGFR1 protein and mRNA levels were decreased in histological sections of the parathyroid glands. Recombinant FGF23 failed to decrease serum parathyroid hormone levels or activate the mitogen-activated protein kinase signaling pathway in the glands of rats with advanced experimental chronic kidney disease. In parathyroid gland organ culture, the addition of FGF23 decreased parathyroid hormone secretion and mRNA levels in control animals or rats with early but not advanced chronic kidney disease. Our results show that because of a downregulation of the Klotho-FGFR1c receptor complex, an increase of circulating FGF23 does not decrease parathyroid hormone levels in established chronic kidney disease. This in vivo resistance is sustained in parathyroid organ culture in vitro. PMID- 20016469 TI - Aberrantly glycosylated IgA1 induces mesangial cells to produce platelet activating factor that mediates nephrin loss in cultured podocytes. AB - The reaction of mesangial cells with aberrantly glycosylated IgA1 has been implicated in the etiology of IgA nephropathy (IgAN). Tumor necrosis factor, which is assumed to mediate the interaction between mesangial cells and podocytes, also induces the expression of platelet-activating factor (PAF). In this study, we determined whether PAF affects the expression of nephrin (an adhesion molecule critical to glomerular permselectivity) and cytoskeletal F actin organization in podocytes. We treated human mesangial cells with atypically glycosylated IgA1 either prepared in vitro or derived from the sera of patients with IgAN. We then prepared conditioned media from these cells and added them to cultured human podocytes in the presence of PAF receptor antagonists. Podocytes transfected to overexpress acetylhydrolase, the main catabolic enzyme of PAF, served as controls. Downregulation of nephrin expression and F-actin reorganization occurred when podocytes were cultured with mesangial cell conditioned medium. Preincubation of podocytes with a PAF receptor antagonist prevented the loss and redistribution of nephrin. In control podocytes overexpressing acetylhydrolase, nephrin loss was abrogated. Our results suggest that atypically glycosylated IgA-induced PAF from mesangial cells is a mediator of podocyte changes, which, when more directly tested elsewhere, were found to be associated with proteinuria. Hence, it is possible that these in vitro findings may be relevant to the proteinuria of IgAN. PMID- 20016470 TI - Hypoxia-inducible factor-2alpha-expressing interstitial fibroblasts are the only renal cells that express erythropoietin under hypoxia-inducible factor stabilization. AB - The adaptation of erythropoietin production to oxygen supply is determined by the abundance of hypoxia-inducible factor (HIF), a regulation that is induced by a prolyl hydroxylase. To identify cells that express HIF subunits (HIF-1alpha and HIF-2alpha) and erythropoietin, we treated Sprague-Dawley rats with the prolyl hydroxylase inhibitor FG-4497 for 6 h to induce HIF-dependent erythropoietin transcription. The kidneys were analyzed for colocalization of erythropoietin mRNA with HIF-1alpha and/or HIF-2alpha protein along with cell-specific identification markers. FG-4497 treatment strongly induced erythropoietin mRNA exclusively in cortical interstitial fibroblasts. Accumulation of HIF-2alpha was observed in these fibroblasts and in endothelial and glomerular cells, whereas HIF-1alpha was induced only in tubular epithelia. A large proportion (over 90% in the juxtamedullary cortex) of erythropoietin-expressing cells coexpressed HIF 2alpha. No colocalization of erythropoietin and HIF-1alpha was found. Hence, we conclude that in the adult kidney, HIF-2alpha and erythropoietin mRNA colocalize only in cortical interstitial fibroblasts, which makes them the key cell type for renal erythropoietin synthesis as regulated by HIF-2alpha. PMID- 20016471 TI - Plasma interleukin-6 is independently associated with mortality in both hemodialysis and pre-dialysis patients with chronic kidney disease. AB - Chronic inflammation associated with chronic kidney disease predicts all-cause and cardiovascular mortality in hemodialysis patients. Here we sought to evaluate the association between plasma levels of the inflammatory mediator interleukin-6 (IL-6) and mortality and aortic calcification/stiffness in 125 patients at different stages (2-5D) of chronic kidney disease. Using multivariate linear regression, we found that plasma IL-6 was independently associated with C reactive protein, albumin and the stage of chronic kidney disease, but not the aortic calcification score or pulse wave velocity. During follow-up studies (median of 829 days), 38 patients died, 22 from cardiovascular events. Plasma IL 6 significantly predicted overall and cardiovascular mortality; this association persisted after multiple adjustments or restricting the analysis to pre-dialysis patients. Moreover, IL-6 was a significantly better predictor of mortality than C reactive protein, albumin or tumor necrosis factor-alpha. Hence, plasma IL-6 independently predicted overall and cardiovascular mortality in patients at different stages of chronic kidney disease; however, whether lowering plasma IL-6 will affect the outcome of chronic kidney disease will require more direct evaluation. PMID- 20016472 TI - Dissociation of embryonic kidneys followed by reaggregation allows the formation of renal tissues. AB - Here we describe a novel method in which embryonic kidneys are dissociated into single-cell suspensions and then reaggregated to form organotypic renal structures. Kidney cell reaggregates were transiently cultured with small molecule Rho kinase inhibitors, which caused ureteric bud structures to form and induced formation of nephrons. These structures displayed normal morphology, expressed appropriate differentiation markers, and were connected at their distal ends to the ureteric buds, thus forming artificial tissues very similar to those found in normal embryonic kidneys. Using this culture method, it was straightforward to make fine-grained chimeras by mixing different cell types or by mixing cells transfected with different constructs before reaggregation. Chimeric renal cultures were formed using mixtures of unmarked normal host embryonic kidney cells and CellTracker-marked WT1 siRNA-carrying cells to test the hypothesis that WT1 is important to a cell's ability to contribute to nephron formation. We found a significant reduction in the ability of WT1 knockdown cells to contribute to nephron formation. This dissociation and reaggregation procedure can also be applied to embryonic lungs and to form coarse-grained hybrid tissues from mixtures of lung and kidney cells. Overall, our protocol allows very simple mixing of cells from different sources or cells subjected to different pretreatments to make fine-grained, highly dispersed chimera tissues. PMID- 20016473 TI - Host immune response to antibiotic perturbation of the microbiota. AB - Mammals are superorganisms, being a composite of mammalian and microbial cells existing in symbiosis. Although the microbiota is not essential for life, commensal and intestinal epithelial cell interactions are critical for the maturation of the immune system. Antibiotic treatment alters this delicate balance by causing compositional changes in the intestinal microbiota, and may lead to a homeostatic imbalance through alterations in expression of IEC tight junction proteins, mucin, antimicrobial peptides, and cytokines. Dysregulation of the homeostasis between mammals and their intestinal symbionts has been shown to predispose the host to enteric infection, and may lead to development of inflammatory bowel diseases. PMID- 20016474 TI - The influence of TSLP on the allergic response. AB - Exposure to allergens first occurs at body surfaces in direct contact with the environment such as the skin, airways, and gastrointestinal tract, and compelling evidence suggests that allergic inflammatory responses are profoundly influenced by the products of epithelial cells located at these sites. One such product is thymic stromal lymphopoietin (TSLP), which is capable of affecting multiple cell lineages involved in allergic reactions. In this review we discuss recent work that has provided insight into the role TSLP plays in both aberrant and protective allergic inflammatory responses, as well as regulation, associations with disease, sources, and functions of this important cytokine. PMID- 20016479 TI - Comment on "Gut-associated lymphoid tissue contains the molecular machinery to support T-cell-dependent and T-cell-independent class switch recombination". PMID- 20016481 TI - 'Allogeneic marrow transplantation in children with acute leukemia: a practice whose time has gone': twenty years later. PMID- 20016484 TI - Three-dimensional structure determination from a single view. AB - The ability to determine the structure of matter in three dimensions has profoundly advanced our understanding of nature. Traditionally, the most widely used schemes for three-dimensional (3D) structure determination of an object are implemented by acquiring multiple measurements over various sample orientations, as in the case of crystallography and tomography, or by scanning a series of thin sections through the sample, as in confocal microscopy. Here we present a 3D imaging modality, termed ankylography (derived from the Greek words ankylos meaning 'curved' and graphein meaning 'writing'), which under certain circumstances enables complete 3D structure determination from a single exposure using a monochromatic incident beam. We demonstrate that when the diffraction pattern of a finite object is sampled at a sufficiently fine scale on the Ewald sphere, the 3D structure of the object is in principle determined by the 2D spherical pattern. We confirm the theoretical analysis by performing 3D numerical reconstructions of a sodium silicate glass structure at 2 A resolution, and a single poliovirus at 2-3 nm resolution, from 2D spherical diffraction patterns alone. Using diffraction data from a soft X-ray laser, we also provide a preliminary demonstration that ankylography is experimentally feasible by obtaining a 3D image of a test object from a single 2D diffraction pattern. With further development, this approach of obtaining complete 3D structure information from a single view could find broad applications in the physical and life sciences. PMID- 20016485 TI - A comprehensive catalogue of somatic mutations from a human cancer genome. AB - All cancers carry somatic mutations. A subset of these somatic alterations, termed driver mutations, confer selective growth advantage and are implicated in cancer development, whereas the remainder are passengers. Here we have sequenced the genomes of a malignant melanoma and a lymphoblastoid cell line from the same person, providing the first comprehensive catalogue of somatic mutations from an individual cancer. The catalogue provides remarkable insights into the forces that have shaped this cancer genome. The dominant mutational signature reflects DNA damage due to ultraviolet light exposure, a known risk factor for malignant melanoma, whereas the uneven distribution of mutations across the genome, with a lower prevalence in gene footprints, indicates that DNA repair has been preferentially deployed towards transcribed regions. The results illustrate the power of a cancer genome sequence to reveal traces of the DNA damage, repair, mutation and selection processes that were operative years before the cancer became symptomatic. PMID- 20016486 TI - Coots use hatch order to learn to recognize and reject conspecific brood parasitic chicks. AB - Avian brood parasites and their hosts provide model systems for investigating links between recognition, learning, and their fitness consequences. One major evolutionary puzzle has continued to capture the attention of naturalists for centuries: why do hosts of brood parasites generally fail to recognize parasitic offspring after they have hatched from the egg, even when the host and parasitic chicks differ to almost comic degrees? One prominent theory to explain this pattern proposes that the costs of mistakenly learning to recognize the wrong offspring make recognition maladaptive. Here we show that American coots, Fulica americana, can recognize and reject parasitic chicks in their brood by using learned cues, despite the fact that the hosts and the brood parasites are of the same species. A series of chick cross-fostering experiments confirm that coots use first-hatched chicks in a brood as referents to learn to recognize their own chicks and then discriminate against later-hatched parasitic chicks in the same brood. When experimentally provided with the wrong reference chicks, coots can be induced to discriminate against their own offspring, confirming that the learning errors proposed by theory can exist. However, learning based on hatching order is reliable in naturally parasitized coot nests because host eggs hatch predictably ahead of parasite eggs. Conversely, a lack of reliable information may help to explain why the evolution of chick recognition is not more common in hosts of most interspecific brood parasites. PMID- 20016487 TI - Identification of sister chromatids by DNA template strand sequences. AB - It is generally assumed that sister chromatids are genetically and functionally identical and that segregation to daughter cells is a random process. However, functional differences between sister chromatids regulate daughter cell fate in yeast and sister chromatid segregation is not random in Escherichia coli. Differentiated sister chromatids, coupled with non-random segregation, have been proposed to regulate cell fate during the development of multicellular organisms. This hypothesis has not been tested because molecular features to reliably distinguish between sister chromatids are not obvious. Here we show that parental 'Watson' and 'Crick' DNA template strands can be identified in sister chromatids of murine metaphase chromosomes using CO-FISH (chromosome orientation fluorescence in situ hybridization) with unidirectional probes specific for centromeric and telomeric repeats. All chromosomes were found to have a uniform orientation with the 5' end of the short arm on the same strand as T-rich major satellite repeats. The invariable orientation of repetitive DNA was used to differentially label sister chromatids and directly study mitotic segregation patterns in different cell types. Whereas sister chromatids appeared to be randomly distributed between daughter cells in cultured lung fibroblasts and embryonic stem cells, significant non-random sister chromatid segregation was observed in a subset of colon crypt epithelial cells, including cells outside positions reported for colon stem cells. Our results establish that DNA template sequences can be used to distinguish sister chromatids and follow their mitotic segregation in vivo. PMID- 20016489 TI - Embracing html. PMID- 20016488 TI - A small-cell lung cancer genome with complex signatures of tobacco exposure. AB - Cancer is driven by mutation. Worldwide, tobacco smoking is the principal lifestyle exposure that causes cancer, exerting carcinogenicity through >60 chemicals that bind and mutate DNA. Using massively parallel sequencing technology, we sequenced a small-cell lung cancer cell line, NCI-H209, to explore the mutational burden associated with tobacco smoking. A total of 22,910 somatic substitutions were identified, including 134 in coding exons. Multiple mutation signatures testify to the cocktail of carcinogens in tobacco smoke and their proclivities for particular bases and surrounding sequence context. Effects of transcription-coupled repair and a second, more general, expression-linked repair pathway were evident. We identified a tandem duplication that duplicates exons 3 8 of CHD7 in frame, and another two lines carrying PVT1-CHD7 fusion genes, indicating that CHD7 may be recurrently rearranged in this disease. These findings illustrate the potential for next-generation sequencing to provide unprecedented insights into mutational processes, cellular repair pathways and gene networks associated with cancer. PMID- 20016491 TI - Biochemical networks: the evolution of gene annotation. PMID- 20016492 TI - Synthetic biology: an RNP switch raises a roadblock. PMID- 20016493 TI - Protein folding: sticky N17 speeds huntingtin pile-up. PMID- 20016494 TI - Follow the leader: the use of leader peptides to guide natural product biosynthesis. AB - The avalanche of genomic information in the past decade has revealed that natural product biosynthesis using the ribosomal machinery is much more widespread than originally anticipated. Nearly all of these compounds are crafted through post translational modifications of a larger precursor peptide that often contains the marching orders for the biosynthetic enzymes. We review here the available information for how the peptide sequences in the precursors govern the post translational tailoring processes for several classes of natural products. In addition, we highlight the great potential these leader peptide-directed biosynthetic systems offer for engineering conformationally restrained and pharmacophore-rich products with structural diversity that greatly expands the proteinogenic repertoire. PMID- 20016495 TI - Synthetic translational regulation by an L7Ae-kink-turn RNP switch. AB - The regulation of cell signaling pathways and the reconstruction of genetic circuits are important aspects of bioengineering research. Both of these goals require molecular devices to transmit information from an input biomacromolecule to the desired outputs. Here, we show that an RNA-protein (RNP)-containing L7Ae kink-turn interaction can be used to construct translational regulators under control of an input protein that regulates the expression of desired output proteins. We built a system in which L7Ae, an archaeal ribosomal protein, regulates the translation of a designed mRNA in vitro and in human cells. The translational regulator composed of the RNP might provide new therapeutic strategies based on the detection, repair or rewiring of intrinsic cellular defects, and it may also serve as an invaluable tool for the dissection of the behavior of complex, higher-order circuits in the cell. PMID- 20016496 TI - Motivations for seeking treatment for ED: the woman's perspective. AB - There is increasing evidence that partners have a major role in treatment-seeking behavior for men with ED. This study investigated the motivations of 100 females for seeking medical treatment for their partner's ED. Semi-structured interviews focused on reasons for seeking treatment from the female's perspective. The themes that emerged from the data centered on the importance of sex in the relationship, with closeness and intimacy frequently being seen as more important than sex. The second major theme related to hopes that females had in relation to the medication, particularly in relation to increasing their partner's confidence and reducing his sexual frustration. Enhancement of the relationship as well as improving the female's own feelings of self-doubt and sexual frustration were also mentioned. The results of this study show the multi-faceted nature of the motivations that females express in terms of seeking help for their partner's ED. PMID- 20016497 TI - Replication of LCE3C-LCE3B CNV as a risk factor for psoriasis and analysis of interaction with other genetic risk factors. AB - Recently, a deletion of two late cornified envelope (LCE) genes within the epidermal differentiation complex on chromosome 1 was shown to be overrepresented in 1,426 psoriasis vulgaris (PsV) patients of European ancestry. In this study, we report a confirmation of this finding in 1,354 PsV patients and 937 control individuals of German origin. We found an allele frequency of the deletion of 70.9% in PsV patients and of 64.9% in control individuals (chi(2)=17.44, P=2.97 x 10(-5), odds ratio (95% confidence interval)=1.31 (1.15-1.48)). The overall copy number of the two LCE genes had no influence on the age of onset, but we observed a dosage effect at the genotype level. There was no evidence of statistically significant interaction with copy number of the beta-defensin cluster on 8p23.1 or with an IL-23R pathway variant in a combined data set of German and Dutch individuals, whereas evidence for interaction with the PSORS1 risk allele in German individuals was marginal and did not remain significant after correction for multiple testing. Our study confirms the recently published finding that the deletion of the two LCE genes is a susceptibility factor for PsV with dosage effect, while, because of power limitation, no final conclusion regarding interaction with other PsV risk factors can be made at this stage. PMID- 20016498 TI - Basic fibroblast growth factor regulates persistent ERK oscillations in premalignant but not malignant JB6 cells. AB - The regulation of extracellular signal-regulated kinase (ERK) oscillations in the context of wound healing and carcinogenesis have been investigated in premalignant and malignant JB6 mouse epidermal cells stimulated with basic fibroblast growth factor (bFGF) and 12-O-tetradecanoyl phorbol-13-acetate (TPA). In premalignant JB6 cells, bFGF stimulation (1) increases cellular phospho-ERK and phospho-c-Jun levels, (2) increases serum-dependent cell proliferation, (3) induces an apparent epithelial-to-mesenchymal transition, and (4) induces the persistent nuclear-cytosolic oscillation of an ERK1-green fluorescent protein (ERK1-GFP) chimera. In contrast, TPA induces persistent activation of ERK in the absence of oscillations and does not induce efficient migration. Treatment of malignant or transformed JB6 cells with bFGF is associated with a transient nuclear translocation of ERK1-GFP but not oscillations or efficient cell migration. Our data suggest that bFGF regulates ERK oscillations in premalignant but not malignant JB6 cells. PMID- 20016499 TI - No increased risk of cancer after coal tar treatment in patients with psoriasis or eczema. AB - Coal tar is an effective treatment for psoriasis and eczema, but it contains several carcinogenic compounds. Occupational and animal studies have shown an increased risk of cancer after exposure to coal tar. Many dermatologists have abandoned this treatment for safety reasons, although the risk of cancer after coal tar in dermatological practice is unclear. This large cohort study included 13,200 patients with psoriasis and eczema. Information on skin disease and treatment, risk factors, and cancer occurrence was retrieved from medical files, questionnaires, and medical registries. Proportional hazards regression was used to evaluate differences in cancer risk by treatment modality. Patients treated with coal tar were compared with a reference category of patients treated with dermatocorticosteroids (assumed to carry no increased cancer risk). The median exposure to coal tar ointments was 6 months (range 1-300 months). Coal tar did not increase the risk of non-skin malignancies (hazard ratio (HR) 0.92; 95% confidence interval (CI) 0.78-1.09), or the risk of skin cancer (HR 1.09; 95% CI 0.69-1.72). This study has sufficient power to show that coal tar treatment is not associated with an increased risk of cancer. These results indicate that coal tar can be maintained as a safe treatment in dermatological practice. PMID- 20016500 TI - Inflammatory monocytes are a reservoir for Merkel cell polyomavirus. AB - Merkel cell polyomavirus (MCPyV) is a recently discovered virus that is implicated in the oncogenesis of Merkel cell carcinoma (MCC). The route of dissemination and the reservoir(s) of MCPyV within the human body have not yet been identified. In this study we describe two patients with multiple MCPyV positive inflammatory and neoplastic skin lesions at different anatomic sites. Patient 1 was suffering from psoriasis for many years and was diagnosed with MCC 7 years before this study. Patient 2 had developed numerous non-melanoma skin cancer lesions under post-transplant immunosuppression. In both patients, MCPyV DNA was detected in whole blood and in urine using PCR and direct sequencing of PCR products. When we analyzed different blood compartments, we found MCPyV exclusively in cell-free serum and in blood monocytes, but not in lymphocytes or granulocytes. Upon separate analysis of resident (CD14(lo)CD16(+)) and inflammatory (CD14(+)CD16(-)) monocytes, we detected MCPyV exclusively in inflammatory, but not in resident monocytes. Our findings raise the possibility that MCPyV persists in inflammatory monocytes and spreads along the migration routes of inflammatory monocytes. This points to intervention strategies to contain MCPyV. Moreover, blood or urine tests may serve as ancillary tests to confirm MCPyV infection in a clinical setting. PMID- 20016501 TI - PLEC1 mutations underlie adult-onset dilated cardiomyopathy in epidermolysis bullosa simplex with muscular dystrophy. PMID- 20016502 TI - Lighting the way forward. PMID- 20016503 TI - Levees of immunological tolerance. AB - Immunological tolerance guards against spurious immune responses to body constituents. Tolerance encompasses a network of mechanisms: central and peripheral, cell-autonomous and cell-interactive. Our understanding of these mechanisms has improved greatly over recent years, often reflecting new insights into the processes underlying particular autoimmune diseases. Yet it is possible that important tolerance mechanisms remain to be discovered, perhaps an explanation for the so-far disappointing clinical translation to the prevention and cure of autoimmune diseases. PMID- 20016504 TI - Regulatory T cells exert checks and balances on self tolerance and autoimmunity. AB - Immunological self tolerance is maintained at least in part by regulatory T (T(reg)) cells that actively and dominantly control potentially hazardous self reactive T cells in the periphery. Antigens that stimulate self-reactive T cells may also activate natural T(reg) cells, thereby maintaining dominant self tolerance. Conversely, genetic anomalies or environmental agents that specifically or predominantly affect T(reg) cells cause or predispose to autoimmunity. With recent advances in our understanding of T(reg) cell development in the thymus and periphery and the molecular mechanism of T(reg) cell-mediated suppression, new ways of treating immunological diseases by targeting T(reg) cells at the cellular and molecular levels are envisaged. PMID- 20016505 TI - Checkpoints in lymphocyte development and autoimmune disease. AB - Antigen receptor-controlled checkpoints in B lymphocyte development are crucial for the prevention of autoimmune diseases such as systemic lupus erythematosus. Checkpoints at the stage of pre-B cell receptor (pre-BCR) and BCR expression can eliminate certain autoreactive BCRs either by deletion of or anergy induction in cells expressing autoreactive BCRs or by receptor editing. For T cells, the picture is more complex because there are regulatory T (T(reg)) cells that mediate dominant tolerance, which differs from the recessive tolerance mediated by deletion and anergy. Negative selection of thymocytes may be as essential as T(reg) cell generation in preventing autoimmune diseases such as type 1 diabetes, but supporting evidence is scarce. Here we discuss several scenarios in which failures at developmental checkpoints result in autoimmunity. PMID- 20016506 TI - Mechanisms maintaining peripheral tolerance. AB - The presentation of self-peptide-MHC complexes in the periphery to potentially autoreactive T cells that have escaped negative selection in the thymus poses an important problem to the immune system. In this review, I discuss data that reveal barriers preventing peripheral T cell recognition of self-peptide-MHC complexes, as well as the physiological mechanisms that ensure the elimination or functional inactivation (anergy) of T cells that do come to recognize self peptide-MHC and threaten the health of the individual. PMID- 20016508 TI - Autoimmunity: increasing suspects in the CD4+ T cell lineup. AB - Chronic reactivity of CD4(+) T cells to autoantigens and to components of the commensal flora drive destructive inflammation in a variety of mouse models of autoimmunity. Insight gained using these models is empowering translational research into human disease. Immunologists are trying to assign disease culpability to one of the ever-growing number of T helper (T(H)) cell subsets. Although recent discovery of the interleukin 17-producing T(H)-17 lineage appeared to supplant the pre-eminence of T(H)1 cells in promoting autoimmunity, the newest data defy simple paradigms. Here we speculate on the respective contributions to autoimmunity made by an increasingly complex list of T(H) subsets and argue that the T(H)1 phenotype may be staging a comeback. PMID- 20016507 TI - Influence of microbial environment on autoimmunity. AB - During protective immune responses, the adaptive arm of the immune system requires activation by signals provided by innate immunity and driven by microbial stimuli. Whether the same rules apply to autoimmune diseases involving clonal self-reactive T and B lymphocytes--a process referred to here as 'adaptive autoimmunity'--is not quite clear. Nevertheless, in these diseases, the innate adaptive connection is likely to be influenced by the microbial environment. This review integrates the results of experiments analyzing autoimmunity in sterile versus nonsterile conditions and experiments testing the role of innate immune receptor signaling in autoimmunity. It proposes that autoimmune diseases can be divided into two groups, the pathogenesis of which either follows the rules of innate-adaptive connection or does not. PMID- 20016509 TI - Mixed results with modulation of TH-17 cells in human autoimmune diseases. AB - The outcomes of clinical trials provide the most convincing data to clarify the role of particular cytokines in the pathogenesis of human diseases. The immunology community, for a variety of practical reasons, spends most of its research time and funds on studies in model systems, mainly mice. In this perspective I discuss results of clinical trials assessing the effect of blocking the differentiation and/or function of interleukin-17-producing CD4(+) T cells on human autoimmune disease, and devote more limited attention to corroborating preclinical studies from animal models. Thus far, these outcomes in human trials have been mixed, with notable success in psoriasis and Crohn's disease but a negative result in relapsing-remitting multiple sclerosis. PMID- 20016511 TI - Glimpsing the real CD4+ T cell response. PMID- 20016512 TI - Defensins keep the peace too. PMID- 20016513 TI - Taking T cells beyond the diffraction limit. PMID- 20016520 TI - Spironolactone reduces severity of obstructive sleep apnoea in patients with resistant hypertension: a preliminary report. AB - Obstructive sleep apnoea (OSA) and hyperaldosteronism are very common in subjects with resistant hypertension. We hypothesized that aldosterone-mediated chronic fluid retention may influence OSA severity in patients with resistant hypertension. We tested this in an open-label evaluation by assessing the changes in the severity of OSA in patients with resistant hypertension after treatment with spironolactone. Subjects with resistant hypertension (clinical blood pressure (BP) >or=140/90 mm Hg on >or=3 antihypertensive medications, including a thiazide diuretic and OSA (defined as an apnoea-hypopnoea index (AHI) >or=15) had full diagnostic, polysomnography before and 8 weeks after spironolactone (25-50 mg a day) was added to their ongoing antihypertensive therapy. In all, 12 patients (mean age 56 years and body mass index 36.8 kg m(-2)) were evaluated. After treatment with spironolactone, the AHI (39.8+/-19.5 vs 22.0+/-6.8 events/h; P<0.05) and hypoxic index (13.6+/-10.8 vs 6.7+/-6.6 events/h; P<0.05), weight and clinic and ambulatory BP were significantly reduced. Plasma renin activity (PRA) and serum creatinine were significantly higher. This study provides preliminary evidence that treatment with a mineralocorticoid receptor antagonist substantially reduces the severity of OSA. If confirmed in a randomized assessment, it will support aldosterone-mediated chronic fluid retention as an important mediator of OSA severity in patients with resistant hypertension. PMID- 20016521 TI - Carotid atherosclerosis in elderly hypertensive patients: potential role of endothelin and plasma antioxidant capacity. AB - Endothelin-1 (ET-1) and oxidative stress are involved in the development of hypertension-induced cardiovascular complications. The aim of this study was to evaluate the relationship between plasma ET-1 level and plasma antioxidant capacity and carotid atherosclerosis. In 61 treated patients with hypertension (44 women, 35 diabetics, mean age 72.4+/-7.2 years) medical histories, ambulatory blood pressure, blood tests (glucose, creatinine, cholesterol, haemoglobin A1c (HbA1c), ET-1) and common carotid artery intima-media thickness (CCA-IMT) measurement were carried out. Plasma antioxidant capacity was assessed by the ferric-reducing ability of plasma (FRAP). Subjects with diabetes presented with higher concentrations of glucose (7.01+/-2.3 vs 5.14+/-0.6 mmol l(-1), P<0.001), HbA1c (7.75+/-2.1 vs 6.1+/-1.2%, P<0.001) and ET-1 (1.36+/-0.53 vs 1.01+/-0.4 pg ml(-1), P<0.01), and lower cholesterol level (5.02+/-0.8 vs 5.86+/-1.3 mmol l( 1), P<0.01). A significant positive correlation between CCA-IMT and ET-1 plasma concentration (r=0.40, P<0.001) and reverse relationship between CCA-IMT and FRAP (r=-0.36, P<0.01) was observed. In a stepwise regression analysis, after adjustment for all confounders, CCA-IMT was independently influenced by age, systolic blood pressure (SBP), HbA1c and ET-1. When FRAP was included in the regression model, CCA-IMT was significantly influenced by age, FRAP, HbA1c and SBP. ET-1 promotes the increase in CCA-IMT contributing to the development of end organ damage. Plasma antioxidant capacity may modulate this deleterious effect, but whether better antioxidant defence may prevent against the development of atherosclerosis remains to be elucidated. PMID- 20016522 TI - Total arterial compliance: the future of cardiovascular risk assessment? PMID- 20016523 TI - Comparison of the efficacy of candesartan and losartan: a meta-analysis of trials in the treatment of hypertension. AB - Informed by the findings from prospective observational studies and randomized outcome trials, guidelines for the management of hypertension acknowledge that the benefit of treatment can be attributed largely to blood pressure (BP) reduction. Therefore, quantification of differential BP lowering of different agents within classes of anti-hypertensives is of practical importance. The objective of this analysis was to compare the efficacy of candesartan and losartan with respect to reduction in systolic and diastolic BP (SBP and DBP). A systematic literature search of databases from 1980 to 1 October 2008 identified 13 studies in which candesartan and losartan were compared in randomized trials in hypertensive patients. Data from 4066 patients were included in the analysis using a random effect model. Mean changes in SBP and DBP were compared for each drug alone and after stratification for dose and for combination with hydrochlorothiazide (HCTZ). On the basis of all the data, the weighted mean difference favoured candesartan-3.22 mm Hg (95% confidence interval (CI) 2.16, 4.29) for SBP and 2.21 mm Hg (95% CI 1.34, 3.07) for DBP. These findings were consistent when analyses according to dose and combination with HCTZ were carried out. Thus, it can be concluded that at currently recommended doses, candesartan is more effective than losartan in lowering BP. PMID- 20016524 TI - Mid-life blood pressure levels and the 8-year incidence of type 2 diabetes mellitus: the Rancho Bernardo Study. AB - Type 2 diabetes mellitus (T2DM) and hypertension frequently occur together. We examined whether blood pressure (BP) levels predict 8-year incident diabetes. Participants were community-dwelling older adults who had BP measured twice and an oral glucose tolerance test at baseline and again 8.3 years later. At baseline, participants were classified as normotensive (systolic blood pressure (SBP) <120 mm Hg and diastolic blood pressure (DBP) <80 mm Hg; n=242); prehypertensive (SBP>or=120 and <140 mm Hg or DBP>or=80 and <90 mm Hg; n=426); or hypertensive (SBP>or=140 mm Hg or DBP>or=90 mm Hg or using anti-hypertensive medication; n=457). There were 1125 participants (mean age 66.0 years; 44.3% men) who attended the baseline and follow-up visit, of whom 85 had new onset T2DM. Participants who developed T2DM had higher mean body mass index (BMI) and BP levels than those who did not develop diabetes. In logistic regression models adjusted for age, sex, BMI, and physical activity, the odds of incident T2DM was greater in prehypertensives (odds ratio (OR) 2.32 95% confidence interval (CI) 1.05-5.1, P=0.03) and hypertensives (OR 3.5 95% CI 1.50-8.0, P=0.002) compared with normotensives. Excluding participants who used anti-hypertensive medications did not change results. In conclusion, mid-life hypertension and prehypertension predicted future diabetes, independent of BMI. Glucose surveillance should be encouraged in adults with prehypertension or hypertension. PMID- 20016525 TI - Identical pattern of cerebral hypoperfusion during different types of syncope. AB - Syncope is caused by insufficient oxygen supply to the brain. There have been attempts to classify syncope on the basis of defects in the venous system, arterial system (that is impaired systemic vascular resistance) or a combination of the two (that is mixed). We examined the hypothesis that a comparable decrease in cerebral perfusion would be evident at pre-syncope irrespective of the category of dysfunction. Young healthy volunteers (N=37) participated. The protocol consisted of 15 min supine rest, followed by 60 degrees head-up tilt and lower body suction in increments of -10 mm Hg for 5 min each until pre-syncope. Beat-to-beat blood pressure (BP) (Finometer or intra-arterial), cardiac output (Finometer), middle cerebral artery blood velocity (MCAv), end-tidal CO(2) and cerebral oxygenation were monitored continuously. At pre-syncope, mixed dysfunction was common (21 out of 37 participants), followed by venular dysfunction (15 out of 37 participants). In the venular and mixed groups, comparable orthostatic tolerance and declines in BP (-37 vs -43% from baseline, respectively), end-tidal PCO(2), MCAv (-35 vs -38%) and cerebral oxygenation (-5 vs -7%) were evident despite distinct mechanisms purportedly being responsible for the hypotension. Although different determinants of hypotension do exist, cerebral hypoperfusion occurs to a similar extent. PMID- 20016526 TI - Arterial stiffening in hypertension: beyond blood pressure levels. PMID- 20016528 TI - Long-term results of Dutch Childhood Oncology Group studies for children with acute lymphoblastic leukemia from 1984 to 2004. AB - The Dutch Childhood Oncology Group (DCOG) has used two treatment strategies for children with acute lymphoblastic leukemia (ALL) based on Pinkel's St Jude Total Therapy or the Berlin-Frankfurt-Munster (BFM) backbone. In four successive protocols, 1734 children were treated. Studies ALL-6 and ALL-9 followed the Total Therapy approach; cranial irradiation was replaced by medium-dose methotrexate infusions and prolonged triple intrathecal therapy; dexamethasone was used instead of prednisone. Studies ALL-7 and ALL-8 had a BFM backbone, including more intensive remission induction, early reinduction and maintenance therapy without vincristine and prednisone pulses. The 5-year event-free survival and overall survival increased from 65.4 to 80.6% (P<0.001) and from 78.7 to 86.4% (P=0.07) in ALL-7 and ALL-9, respectively. In ALL-7 and ALL-8 National Cancer Institute (NCI) high-risk criteria, male gender, T-lineage ALL and high white blood cells (WBCs) predict poor outcome. In ALL-9 NCI criteria, gender, WBC >100 x 109/l, and T-lineage ALL have prognostic impact. We conclude that the chemotherapy-only approach in children with ALL in Total Therapy-based strategies and BFM-backbone treatment does not jeopardize survival and preserves cognitive functioning. This experience is implemented in the current DCOG-ALL-10 study using a BFM backbone and minimal residual disease-based stratification. PMID- 20016527 TI - Long-term results of the pediatric oncology group studies for childhood acute lymphoblastic leukemia 1984-2001: a report from the children's oncology group. AB - From 1984 to 2001, the Pediatric Oncology Group (POG) conducted 12 acute lymphoblastic leukemia (ALL) studies. Ten-year event-free survival (EFS) for patients >12 months of age with B-precursor ALL on acute leukemia in children 14, 15 and 16 series were 66.7+/-1.2%, 68.1+/-1.4% and 73.2+/-2.1%, respectively. Intermediate dose methotrexate (ID MTX; 1 g/m(2)) improved outcomes for standard risk patients (10-year EFS 77.5+/-2.7% vs 66.3+/-3.1% for oral MTX). Neither MTX intensification (2.5 g/m(2)) nor addition of cytosine arabinoside/daunomycin/teniposide improved outcomes for higher risk patients. Intermediate dose mercaptopurine (1 g/m(2)) failed to improve outcomes for either group. Ten-year EFS for patients with T-cell ALL, POG 8704 and 9404 were 49.1+/ 3.1% and 72.2+/-4.7%, respectively. Intensive asparaginase (10-year EFS 61.8 vs 42.7%) and high-dose MTX (5 g/m(2)) (10-year EFS 78.0 vs 65.8%) improved outcomes. There was a non-significant improvement in EFS for infants (10-year EFS 17.7+/-7.2-31.9+/-8.3%). Prognostic indicators for B-precursor ALL were age and WBC at diagnosis, gender, central nervous system disease, DNA index and cytogenetic abnormalities. Only gender was prognostic in T-cell ALL. In infants, WBC and MLL translocation were linked to inferior outcome. PMID- 20016529 TI - Temporal changes in the incidence and pattern of central nervous system relapses in children with acute lymphoblastic leukaemia treated on four consecutive Medical Research Council trials, 1985-2001. AB - Despite the success of contemporary treatment protocols in childhood acute lymphoblastic leukaemia (ALL), relapse within the central nervous system (CNS) remains a challenge. To better understand this phenomenon, we have analysed the changes in incidence and pattern of CNS relapses in 5564 children enrolled in four successive Medical Research Council-ALL trials between 1985 and 2001. Changes in the incidence and pattern of CNS relapses were examined and the relationship with patient characteristics was assessed. The factors affecting outcome after relapse were determined. Overall, relapses declined by 49%. Decreases occurred primarily in non-CNS and combined relapses with a progressive shift towards later (> or =30 months from diagnosis) relapses (P<0.0001). Although isolated CNS relapses declined, the proportional incidence and timing of relapse remained unchanged. Age and presenting white blood cell (WBC) count were risk factors for CNS relapse. On multivariate analysis, the time to relapse and the trial period influenced outcomes after relapse. Relapse trends differed within biological subtypes. In ETV6-RUNX1 ALL, relapse patterns mirrored overall trends whereas in high hyperdiploidy (HH) ALL, these seem to have plateaued over the latter two trial periods. Intensive systemic and intrathecal chemotherapy have decreased the overall CNS relapse rates and changed the patterns of recurrence. The heterogeneity of therapeutic response in the biological subtypes suggests room for further optimization using currently available chemotherapy. PMID- 20016530 TI - Cooperative study group for childhood acute lymphoblastic leukaemia (COALL): long term results of trials 82,85,89,92 and 97. AB - In this study, the long-term outcome of 1818 patients treated in five consecutive clinical trials (the cooperative study group for childhood acute lymphoblastic leukaemia (COALL) 82, 85, 89, 92 and 97) from 24 cooperating centres in Germany is reported. The probability of event-free survival (pEFS) improved significantly from the first two trials conducted in the 1980s (COALL 82 and COALL 85) to the three trials conducted in the 1990s (COALL 89, 92 and 97) (P=0.001). Through all COALL studies, age > or =10 years and initial white blood cell count (WBC) > or =50 x 10(9)/l and pro-B immunophenotype were of significant prognostic relevance. A refinement of risk assessment has been achieved by in vitro drug sensitivity testing in COALL 92 and 97. In patients with very sensitive leukaemic cells, therapy could be reduced without loss of efficacy. In COALL 97, a further improvement in risk stratification was gained by the molecular assessment of minimal residual disease (MRD) under treatment, which proved to have a superior prognostic effect when compared with in vitro drug sensitivity testing. Importantly, the gradual reduction in central nervous system (CNS) irradiation led to a decreased incidence of brain tumours as a second malignancy. In general, the prevention of treatment-related late effects will be one of the major issues in future studies. It remains to be shown whether prolonged infusions of anthracyclines, which have been implemented into the COALL studies after equal efficacy compared with short-time infusions was confirmed, will be associated with fewer cardiac late effects. Another way to prevent late effects may be a more refined risk assessment allowing for a reduction in cumulative treatment burden. A great challenge in the future will be to improve the overall treatment results, which very likely can only be achieved by the identification of molecularly defined subgroups to which novel, rational therapeutic strategies can be applied. PMID- 20016531 TI - Long-term results of the children's cancer group studies for childhood acute lymphoblastic leukemia 1983-2002: a Children's Oncology Group Report. AB - The Children's Cancer Group enrolled 13 298 young people age <21 years on 1 of 16 protocols between 1983 and 2002. Outcomes were examined in three time periods, 1983-1988, 1989-1995, 1996-2002. Over the three intervals, 10-year event-free survival (EFS) for Rome/National Cancer Institute standard risk (SR) and higher risk (HR) B-precursor patients was 68 and 58%, 77 and 63%, and 78 and 67%, respectively, whereas for SR and HR T-cell patients, EFS was 65 and 56%, 78 and 68%, and 70 and 72%, respectively. Five-year EFS for infants was 36, 38, and 43%, respectively. Seminal randomized studies led to a number of important findings. Stronger post-induction intensification improved outcome for both SR and HR patients. With improved systemic therapy, additional intrathecal (IT) methotrexate effectively replaced cranial radiation. For SR patients receiving three-drug induction, iso-toxic substitution of dexamethasone for prednisone improved EFS. Pegylated asparaginase safely and effectively replaced native asparaginase. Thus, rational therapy modifications yielded better outcomes for both SR and HR patients. These trials provide the platforms for current Children's Oncology Group trials. PMID- 20016532 TI - Mutant Wilms' tumor 1 (WT1) mRNA with premature termination codons in acute myeloid leukemia (AML) is sensitive to nonsense-mediated RNA decay (NMD). PMID- 20016533 TI - Identification of acquired copy number alterations and uniparental disomies in cytogenetically normal acute myeloid leukemia using high-resolution single nucleotide polymorphism analysis. AB - Recent advances in genome-wide single-nucleotide polymorphism (SNP) analyses have revealed previously unrecognized microdeletions and uniparental disomy (UPD) in a broad spectrum of human cancers. As acute myeloid leukemia (AML) represents a genetically heterogeneous disease, this technology might prove helpful, especially for cytogenetically normal AML (CN-AML) cases. Thus, we performed high resolution SNP analyses in 157 adult cases of CN-AML. Regions of acquired UPDs were identified in 12% of cases and in the most frequently affected chromosomes, 6p, 11p and 13q. Notably, acquired UPD was invariably associated with mutations in nucleophosmin 1 (NPM1) or CCAAT/enhancer binding protein-alpha (CEBPA) that impair hematopoietic differentiation (P=0.008), suggesting that UPDs may preferentially target genes that are essential for proliferation and survival of hematopoietic progenitors. Acquired copy number alterations (CNAs) were detected in 49% of cases with losses found in two or more cases affecting, for example, chromosome bands 3p13-p14.1 and 12p13. Furthermore, we identified two cases with a cryptic t(6;11) as well as several non-recurrent aberrations pointing to leukemia-relevant regions. With regard to clinical outcome, there seemed to be an association between UPD 11p and UPD 13q cases with overall survival. These data show the potential of high-resolution SNP analysis for identifying genomic regions of potential pathogenic and clinical relevance in AML. PMID- 20016534 TI - Long-term results of the Israeli National Studies in childhood acute lymphoblastic leukemia: INS 84, 89 and 98. PMID- 20016535 TI - Long-term results of treatment of childhood acute lymphoblastic leukemia in the Czech Republic. PMID- 20016536 TI - Long-term results of the Italian Association of Pediatric Hematology and Oncology (AIEOP) Studies 82, 87, 88, 91 and 95 for childhood acute lymphoblastic leukemia. AB - We analyzed the long-term outcome of 4865 patients treated in Studies 82, 87, 88, 91 and 95 for childhood acute lymphoblastic leukemia (ALL) of the Italian Association of Pediatric Hematology and Oncology (AIEOP). Treatment was characterized by progressive intensification of systemic therapy and reduction of cranial radiotherapy. A progressive improvement of results with reduction of isolated central nervous system relapse rate was obtained. Ten-year event-free survival increased from 53% in Study 82 to 72% in Study 95, whereas survival improved from 64 to 82%. Since 1991, all patients were treated according to Berlin-Frankfurt-Muenster (BFM) ALL treatment strategy. In Study 91, reduced treatment intensity (25%) yielded inferior results, but intensification of maintenance with high-dose (HD)-L-asparaginase (randomized) allowed to compensate for this disadvantage; in high-risk patients (HR, 15%), substitution of intensive polychemotherapy blocks for conventional BFM backbone failed to improve results. A marked improvement of results was obtained in HR patients when conventional BFM therapy was intensified with three polychemotherapy blocks and double delayed intensification (Study 95). The introduction of minimal residual disease monitoring and evaluation of common randomized questions by AIEOP and BFM groups in the protocol AIEOP-BFM-ALL 2000 are expected to further ameliorate treatment of children with ALL. PMID- 20016537 TI - Long-term results of Dana-Farber Cancer Institute ALL Consortium protocols for children with newly diagnosed acute lymphoblastic leukemia (1985-2000). AB - The Dana-Farber Cancer Institute (DFCI) acute lymphoblastic leukemia (ALL) Consortium has been conducting multi-institutional clinical trials in childhood ALL since 1981. The treatment backbone has included 20-30 consecutive weeks of asparaginase during intensification and frequent vincristine/corticosteroid pulses during the continuation phase. Between 1985 and 2000, 1457 children aged 0 18 years were treated on four consecutive protocols: 85-01 (1985-1987), 87-01 (1987-1991), 91-01 (1991-1955) and 95-01 (1996-2000). The 10-year event-free survival (EFS)+/-s.e. by protocol was 77.9+/-2.8% (85-01), 74.2+/-2.3% (87-01), 80.8+/-2.1% (91-01) and 80.5+/-1.8% (95-01). Approximately 82% of patients treated in the 1980s and 88% treated in the 1990s were long-term survivors. Both EFS and overall survival (OS) rates were significantly higher for patients treated in the 1990s compared with the 1980s (P=0.05 and 0.01, respectively). On the two protocols conducted in the 1990s, EFS was 79-85% for T-cell ALL patients and 75-78% for adolescents (age 10-18 years). Results of randomized studies revealed that dexrazoxane prevented acute cardiac injury without adversely affecting EFS or OS in high-risk (HR) patients, and frequently dosed intrathecal chemotherapy was an effective substitute for cranial radiation in standard-risk (SR) patients. Current studies continue to focus on improving efficacy while minimizing acute and late toxicities. PMID- 20016538 TI - Long-term results of Taiwan Pediatric Oncology Group studies 1997 and 2002 for childhood acute lymphoblastic leukemia. AB - The long-term outcome of 1390 children with acute lymphoblastic leukemia (ALL), treated in two successive clinical trials (Taiwan Pediatric Oncology Group (TPOG) ALL-97 and TPOG-ALL-2002) between 1997 and 2007, is reported. The event-free survival improved significantly (P=0.0004) over this period, 69.3+/-1.9% in 1997 2001 to 77.4+/-1.7% in 2002-2007. A randomized trial in TPOG-97 testing L asparaginase versus epidoxorubicin in combination with vincristine and prednisolone for remission induction in standard-risk (SR; low-risk) patients yielded similar outcomes. Another randomized trial, in TPOG-2002, showed that for SR patients, two reinduction courses did not improve long-term outcome over one course. Decreasing use of prophylactic cranial irradiation in the period 1997 2008 was not associated with increased rates of CNS relapse, prompting complete omission of prophylactic cranial irradiation from TPOG protocols, beginning in 2009. Decreased use of etoposide and cranial irradiation likely contributed to the low incidence of second cancers. High-risk B-lineage ALL, T-cell, CD10 negativity, t(9;22), infant, and higher leukocyte count were consistently adverse factors, whereas hyperdiploidy >50 was a consistently favorable factor. Higher leukocyte count and t(9;22) retained prognostic significance in both TPOG-97 and TPOG-2002 by multivariate analysis. Although long-term outcome in TPOG clinical trials is comparable with results being reported worldwide, the persistent strength of certain prognostic variables and the lower frequencies of favorable outcome predictors, such as ETV6-RUNX1 and hyperdiploidy >50, in Taiwanese children warrant renewed effort to cure a higher proportion of patients while preserving their quality of life. PMID- 20016539 TI - Long-term results of the Japanese Childhood Cancer and Leukemia Study Group studies 811, 841, 874 and 911 on childhood acute lymphoblastic leukemia. AB - We analyzed the long-term outcomes of 1021 patients with acute lymphoblastic leukemia (ALL), enrolled in four successive clinical trials (ALL811, ALL841, ALL874 and ALL911) between 1981 and 1993. All patients received risk-adopted therapy according to leukocyte count and age at the time of diagnosis. The median follow-up durations of the four studies were 17.8 years in ALL811, 15.5 years in ALL841, 11.9 years in ALL874 and 15.8 years in ALL911. Patients' event-free survival (EFS) and overall survival (OS) rates at 12 years were 41.0 and 54.3% in ALL811, 50.2 and 60.2% in ALL841, 57.3 and 64.7% in ALL874, and 63.4 and 71.7% in ALL911, respectively. Thus, cure can become a reality for about 70% of children with ALL. There is, however, still a significant difference in survival outcomes according to risk group. Late effects were observed in 70 patients out of 834 (8.4%); hepatitis and short stature were most commonly reported. Reduction of late adverse effects for all patients and development of new treatment strategies for very-high-risk patients are major issues for upcoming trials to address. PMID- 20016541 TI - Reply to Bridges. PMID- 20016540 TI - Single-cycle viral gene expression, rather than progressive replication and oncolysis, is required for VSV therapy of B16 melanoma. AB - A fully intact immune system would be expected to hinder the efficacy of oncolytic virotherapy by inhibiting viral replication. Simultaneously, however, it may also enhance antitumor therapy through initiation of proinflammatory, antiviral cytokine responses at the tumor site. The aim of this study was to investigate the role of a fully intact immune system on the antitumor efficacy of an oncolytic virus. In this respect, injection of oncolytic vesicular stomatitis virus (VSV) into subcutaneous B16ova melanomas in C57Bl/6 mice leads to tumor regression, but it is not associated with viral replicative burst in the tumor. In contrast, intratumoral delivery of VSV induces an acute proinflammatory reaction, which quickly resolves concomitantly with virus clearance. Consistent with the hypothesis that therapy may not be dependent on the ability of VSV to undergo progressive rounds of replication, a single-cycle VSV is equally effective as a fully replication-competent VSV, whereas inactivated viruses do not generate therapy. Even though therapy is dependent on host CD8+ and natural killer cells, these effects are not associated with interferon-gamma-dependent responses against either the virus or tumor. There is, however, a strong correlation between viral gene expression, induction of proinflammatory reaction in the tumor and in vivo therapy. Overall, our results suggest that acute innate antiviral immune response, which rapidly clears VSV from B16ova tumors, is associated with the therapy observed in this model. Therefore, the antiviral immune response to an oncolytic virus mediates an intricate balance between safety, restriction of oncolysis and, potentially, significant immune-mediated antitumor therapy. PMID- 20016542 TI - Lentiviral gene ontology (LeGO) vectors equipped with novel drug-selectable fluorescent proteins: new building blocks for cell marking and multi-gene analysis. AB - Vector-encoded fluorescent proteins (FPs) facilitate unambiguous identification or sorting of gene-modified cells by fluorescence-activated cell sorting (FACS). Exploiting this feature, we have recently developed lentiviral gene ontology (LeGO) vectors (www.LentiGO-Vectors.de) for multi-gene analysis in different target cells. In this study, we extend the LeGO principle by introducing 10 different drug-selectable FPs created by fusing one of the five selection marker (protecting against blasticidin, hygromycin, neomycin, puromycin and zeocin) and one of the five FP genes (Cerulean, eGFP, Venus, dTomato and mCherry). All tested fusion proteins allowed both fluorescence-mediated detection and drug-mediated selection of LeGO-transduced cells. Newly generated codon-optimized hygromycin- and neomycin-resistance genes showed improved expression as compared with their ancestors. New LeGO constructs were produced at titers >10(6) per ml (for non concentrated supernatants). We show efficient combinatorial marking and selection of various cells, including mesenchymal stem cells, simultaneously transduced with different LeGO constructs. Inclusion of the cytomegalovirus early enhancer/chicken beta-actin promoter into LeGO vectors facilitated robust transgene expression in and selection of neural stem cells and their differentiated progeny. We suppose that the new drug-selectable markers combining advantages of FACS and drug selection are well suited for numerous applications and vector systems. Their inclusion into LeGO vectors opens new possibilities for (stem) cell tracking and functional multi-gene analysis. PMID- 20016546 TI - Mind the gap. PMID- 20016547 TI - Credit where credit is due. PMID- 20016548 TI - A class of their own. PMID- 20016556 TI - Journal club. A cancer researcher ponders a fundamental connection between nutrients and gene expression. PMID- 20016562 TI - Budget win for climate probe. PMID- 20016564 TI - Royal Institution faces cash crisis. PMID- 20016565 TI - UK research funding proposal is 'irresponsible'. PMID- 20016566 TI - Satellites beam in biomass estimates. PMID- 20016567 TI - Hope for Japan's key projects. PMID- 20016568 TI - Modellers claim wars are predictable. PMID- 20016569 TI - Consent issue dogs stem-cell approval. PMID- 20016570 TI - French research wins huge cash boost. PMID- 20016571 TI - Genetics: Watching science at work. PMID- 20016573 TI - World view: Out of service. PMID- 20016572 TI - Human genomics: The genome finishers. PMID- 20016574 TI - UK defence group's structure could limit its usefulness. PMID- 20016575 TI - Opening dialogue between the recent and the long ago. PMID- 20016576 TI - Iran's scientists condemn instances of plagiarism. PMID- 20016577 TI - Geothermal quake risks must be faced. PMID- 20016582 TI - Extrasolar planets: Water world larger than Earth. PMID- 20016583 TI - DNA replication: prime-time looping. PMID- 20016585 TI - Global change: Interglacial and future sea level. PMID- 20016586 TI - DNA repair: A heavyweight joins the fray. PMID- 20016587 TI - Nanotechnology: Soggy origami. PMID- 20016588 TI - Neuroscience: New tricks and old spines. PMID- 20016589 TI - Microscopy: Photons and electrons team up. PMID- 20016590 TI - Obituary: Claude Levi-Strauss (1908-2009). PMID- 20016591 TI - Probabilistic assessment of sea level during the last interglacial stage. AB - With polar temperatures approximately 3-5 degrees C warmer than today, the last interglacial stage (approximately 125 kyr ago) serves as a partial analogue for 1 2 degrees C global warming scenarios. Geological records from several sites indicate that local sea levels during the last interglacial were higher than today, but because local sea levels differ from global sea level, accurately reconstructing past global sea level requires an integrated analysis of globally distributed data sets. Here we present an extensive compilation of local sea level indicators and a statistical approach for estimating global sea level, local sea levels, ice sheet volumes and their associated uncertainties. We find a 95% probability that global sea level peaked at least 6.6 m higher than today during the last interglacial; it is likely (67% probability) to have exceeded 8.0 m but is unlikely (33% probability) to have exceeded 9.4 m. When global sea level was close to its current level (>or=-10 m), the millennial average rate of global sea level rise is very likely to have exceeded 5.6 m kyr(-1) but is unlikely to have exceeded 9.2 m kyr(-1). Our analysis extends previous last interglacial sea level studies by integrating literature observations within a probabilistic framework that accounts for the physics of sea level change. The results highlight the long-term vulnerability of ice sheets to even relatively low levels of sustained global warming. PMID- 20016593 TI - Growth landscape formed by perception and import of glucose in yeast. AB - An important challenge in systems biology is to quantitatively describe microbial growth using a few measurable parameters that capture the essence of this complex phenomenon. Two key events at the cell membrane-extracellular glucose sensing and uptake-initiate the budding yeast's growth on glucose. However, conventional growth models focus almost exclusively on glucose uptake. Here we present results from growth-rate experiments that cannot be explained by focusing on glucose uptake alone. By imposing a glucose uptake rate independent of the sensed extracellular glucose level, we show that despite increasing both the sensed glucose concentration and uptake rate, the cell's growth rate can decrease or even approach zero. We resolve this puzzle by showing that the interaction between glucose perception and import, not their individual actions, determines the central features of growth, and characterize this interaction using a quantitative model. Disrupting this interaction by knocking out two key glucose sensors significantly changes the cell's growth rate, yet uptake rates are unchanged. This is due to a decrease in burden that glucose perception places on the cells. Our work shows that glucose perception and import are separate and pivotal modules of yeast growth, the interaction of which can be precisely tuned and measured. PMID- 20016594 TI - The SUMO modification pathway is involved in the BRCA1 response to genotoxic stress. AB - Mutations in BRCA1 are associated with a high risk of breast and ovarian cancer. BRCA1 participates in the DNA damage response and acts as a ubiquitin ligase. However, its regulation remains poorly understood. Here we report that BRCA1 is modified by small ubiquitin-like modifier (SUMO) in response to genotoxic stress, and co-localizes at sites of DNA damage with SUMO1, SUMO2/3 and the SUMO conjugating enzyme Ubc9. PIAS SUMO E3 ligases co-localize with and modulate SUMO modification of BRCA1, and are required for BRCA1 ubiquitin ligase activity in cells. In vitro SUMO modification of the BRCA1/BARD1 heterodimer greatly increases its ligase activity, identifying it as a SUMO-regulated ubiquitin ligase (SRUbL). Further, PIAS SUMO ligases are required for complete accumulation of double-stranded DNA (dsDNA) damage-repair proteins subsequent to RNF8 accrual, and for proficient double-strand break repair. These data demonstrate that the SUMOylation pathway plays a significant role in mammalian DNA damage response. PMID- 20016592 TI - Parental origin of sequence variants associated with complex diseases. AB - Effects of susceptibility variants may depend on from which parent they are inherited. Although many associations between sequence variants and human traits have been discovered through genome-wide associations, the impact of parental origin has largely been ignored. Here we show that for 38,167 Icelanders genotyped using single nucleotide polymorphism (SNP) chips, the parental origin of most alleles can be determined. For this we used a combination of genealogy and long-range phasing. We then focused on SNPs that associate with diseases and are within 500 kilobases of known imprinted genes. Seven independent SNP associations were examined. Five-one with breast cancer, one with basal-cell carcinoma and three with type 2 diabetes-have parental-origin-specific associations. These variants are located in two genomic regions, 11p15 and 7q32, each harbouring a cluster of imprinted genes. Furthermore, we observed a novel association between the SNP rs2334499 at 11p15 and type 2 diabetes. Here the allele that confers risk when paternally inherited is protective when maternally transmitted. We identified a differentially methylated CTCF-binding site at 11p15 and demonstrated correlation of rs2334499 with decreased methylation of that site. PMID- 20016595 TI - A super-Earth transiting a nearby low-mass star. AB - A decade ago, the detection of the first transiting extrasolar planet provided a direct constraint on its composition and opened the door to spectroscopic investigations of extrasolar planetary atmospheres. Because such characterization studies are feasible only for transiting systems that are both nearby and for which the planet-to-star radius ratio is relatively large, nearby small stars have been surveyed intensively. Doppler studies and microlensing have uncovered a population of planets with minimum masses of 1.9-10 times the Earth's mass (M[symbol:see text]), called super-Earths. The first constraint on the bulk composition of this novel class of planets was afforded by CoRoT-7b (refs 8, 9), but the distance and size of its star preclude atmospheric studies in the foreseeable future. Here we report observations of the transiting planet GJ 1214b, which has a mass of 6.55M[symbol:see text]), and a radius 2.68 times Earth's radius (R[symbol:see text]), indicating that it is intermediate in stature between Earth and the ice giants of the Solar System. We find that the planetary mass and radius are consistent with a composition of primarily water enshrouded by a hydrogen-helium envelope that is only 0.05% of the mass of the planet. The atmosphere is probably escaping hydrodynamically, indicating that it has undergone significant evolution during its history. The star is small and only 13 parsecs away, so the planetary atmosphere is amenable to study with current observatories. PMID- 20016596 TI - A single sub-kilometre Kuiper belt object from a stellar occultation in archival data. AB - The Kuiper belt is a remnant of the primordial Solar System. Measurements of its size distribution constrain its accretion and collisional history, and the importance of material strength of Kuiper belt objects. Small, sub-kilometre sized, Kuiper belt objects elude direct detection, but the signature of their occultations of background stars should be detectable. Observations at both optical and X-ray wavelengths claim to have detected such occultations, but their implied abundances are inconsistent with each other and far exceed theoretical expectations. Here we report an analysis of archival data that reveals an occultation by a body with an approximately 500-metre radius at a distance of 45 astronomical units. The probability of this event arising from random statistical fluctuations within our data set is about two per cent. Our survey yields a surface density of Kuiper belt objects with radii exceeding 250 metres of 2.1( 1.7)(+4.8) x 10(7) deg(-2), ruling out inferred surface densities from previous claimed detections by more than 5sigma. The detection of only one event reveals a deficit of sub-kilometre-sized Kuiper belt objects compared to a population extrapolated from objects with radii exceeding 50 kilometres. This implies that sub-kilometre-sized objects are undergoing collisional erosion, just like debris disks observed around other stars. PMID- 20016597 TI - Photon-by-photon feedback control of a single-atom trajectory. AB - Feedback is one of the most powerful techniques for the control of classical systems. An extension into the quantum domain is desirable as it could allow the production of non-trivial quantum states and protection against decoherence. The difficulties associated with quantum, as opposed to classical, feedback arise from the quantum measurement process-in particular the quantum projection noise and the limited measurement rate-as well as from quantum fluctuations perturbing the evolution in a driven open system. Here we demonstrate real-time feedback control of the motion of a single atom trapped in an optical cavity. Individual probe photons carrying information about the atomic position activate a dipole laser that steers the atom on timescales 70 times shorter than the atom's oscillation period in the trap. Depending on the specific implementation, the trapping time is increased by a factor of more than four owing to feedback cooling, which can remove almost all the kinetic energy of the atom in a quarter of an oscillation period. Our results show that the detected photon flux reflects the atomic motion, and thus mark a step towards the exploration of the quantum trajectory of a single atom at the standard quantum limit. PMID- 20016598 TI - Photon-induced near-field electron microscopy. AB - In materials science and biology, optical near-field microscopies enable spatial resolutions beyond the diffraction limit, but they cannot provide the atomic scale imaging capabilities of electron microscopy. Given the nature of interactions between electrons and photons, and considering their connections through nanostructures, it should be possible to achieve imaging of evanescent electromagnetic fields with electron pulses when such fields are resolved in both space (nanometre and below) and time (femtosecond). Here we report the development of photon-induced near-field electron microscopy (PINEM), and the associated phenomena. We show that the precise spatiotemporal overlap of femtosecond single-electron packets with intense optical pulses at a nanostructure (individual carbon nanotube or silver nanowire in this instance) results in the direct absorption of integer multiples of photon quanta (nhomega) by the relativistic electrons accelerated to 200 keV. By energy-filtering only those electrons resulting from this absorption, it is possible to image directly in space the near-field electric field distribution, obtain the temporal behaviour of the field on the femtosecond timescale, and map its spatial polarization dependence. We believe that the observation of the photon-induced near-field effect in ultrafast electron microscopy demonstrates the potential for many applications, including those of direct space-time imaging of localized fields at interfaces and visualization of phenomena related to photonics, plasmonics and nanostructures. PMID- 20016599 TI - Fault zone fabric and fault weakness. AB - Geological and geophysical evidence suggests that some crustal faults are weak compared to laboratory measurements of frictional strength. Explanations for fault weakness include the presence of weak minerals, high fluid pressures within the fault core and dynamic processes such as normal stress reduction, acoustic fluidization or extreme weakening at high slip velocity. Dynamic weakening mechanisms can explain some observations; however, creep and aseismic slip are thought to occur on weak faults, and quasi-static weakening mechanisms are required to initiate frictional slip on mis-oriented faults, at high angles to the tectonic stress field. Moreover, the maintenance of high fluid pressures requires specialized conditions and weak mineral phases are not present in sufficient abundance to satisfy weak fault models, so weak faults remain largely unexplained. Here we provide laboratory evidence for a brittle, frictional weakening mechanism based on common fault zone fabrics. We report on the frictional strength of intact fault rocks sheared in their in situ geometry. Samples with well-developed foliation are extremely weak compared to their powdered equivalents. Micro- and nano-structural studies show that frictional sliding occurs along very fine-grained foliations composed of phyllosilicates (talc and smectite). When the same rocks are powdered, frictional strength is high, consistent with cataclastic processes. Our data show that fault weakness can occur in cases where weak mineral phases constitute only a small percentage of the total fault rock and that low friction results from slip on a network of weak phyllosilicate-rich surfaces that define the rock fabric. The widespread documentation of foliated fault rocks along mature faults in different tectonic settings and from many different protoliths suggests that this mechanism could be a viable explanation for fault weakening in the brittle crust. PMID- 20016600 TI - Common ecology quantifies human insurgency. AB - Many collective human activities, including violence, have been shown to exhibit universal patterns. The size distributions of casualties both in whole wars from 1816 to 1980 and terrorist attacks have separately been shown to follow approximate power-law distributions. However, the possibility of universal patterns ranging across wars in the size distribution or timing of within conflict events has barely been explored. Here we show that the sizes and timing of violent events within different insurgent conflicts exhibit remarkable similarities. We propose a unified model of human insurgency that reproduces these commonalities, and explains conflict-specific variations quantitatively in terms of underlying rules of engagement. Our model treats each insurgent population as an ecology of dynamically evolving, self-organized groups following common decision-making processes. Our model is consistent with several recent hypotheses about modern insurgency, is robust to many generalizations, and establishes a quantitative connection between human insurgency, global terrorism and ecology. Its similarity to financial market models provides a surprising link between violent and non-violent forms of human behaviour. PMID- 20016601 TI - Division and apoptosis of E2f-deficient retinal progenitors. AB - The activating E2f transcription factors (E2f1, E2f2 and E2f3) induce transcription and are widely viewed as essential positive cell cycle regulators. Indeed, they drive cells out of quiescence, and the 'cancer cell cycle' in Rb1 null cells is E2f-dependent. Absence of activating E2fs in flies or mammalian fibroblasts causes cell cycle arrest, but this block is alleviated by removing repressive E2f or the tumour suppressor p53, respectively. Thus, whether activating E2fs are indispensable for normal division is an area of debate. Activating E2fs are also well known pro-apoptotic factors, providing a defence against oncogenesis, yet E2f1 can limit irradiation-induced apoptosis. In flies this occurs through repression of hid (also called Wrinkled; Smac/Diablo in mammals). However, in mammals the mechanism is unclear because Smac/Diablo is induced, not repressed, by E2f1, and in keratinocytes survival is promoted indirectly through induction of DNA repair targets. Thus, a direct pro-survival function for E2f1-3 and/or its relevance beyond irradiation has not been established. To address E2f1-3 function in normal cells in vivo we focused on the mouse retina, which is a relatively simple central nervous system component that can be manipulated genetically without compromising viability and has provided considerable insight into development and cancer. Here we show that unlike fibroblasts, E2f1-3 null retinal progenitor cells or activated Muller glia can divide. We attribute this effect to functional interchangeability with Mycn. However, loss of activating E2fs caused downregulation of the p53 deacetylase Sirt1, p53 hyperacetylation and elevated apoptosis, establishing a novel E2f Sirt1-p53 survival axis in vivo. Thus, activating E2fs are not universally required for normal mammalian cell division, but have an unexpected pro-survival role in development. PMID- 20016602 TI - E2f1-3 switch from activators in progenitor cells to repressors in differentiating cells. AB - In the established model of mammalian cell cycle control, the retinoblastoma protein (Rb) functions to restrict cells from entering S phase by binding and sequestering E2f activators (E2f1, E2f2 and E2f3), which are invariably portrayed as the ultimate effectors of a transcriptional program that commit cells to enter and progress through S phase. Using a panel of tissue-specific cre-transgenic mice and conditional E2f alleles we examined the effects of E2f1, E2f2 and E2f3 triple deficiency in murine embryonic stem cells, embryos and small intestines. We show that in normal dividing progenitor cells E2f1-3 function as transcriptional activators, but contrary to the current view, are dispensable for cell division and instead are necessary for cell survival. In differentiating cells E2f1-3 function in a complex with Rb as repressors to silence E2f targets and facilitate exit from the cell cycle. The inactivation of Rb in differentiating cells resulted in a switch of E2f1-3 from repressors to activators, leading to the superactivation of E2f responsive targets and ectopic cell divisions. Loss of E2f1-3 completely suppressed these phenotypes caused by Rb deficiency. This work contextualizes the activator versus repressor functions of E2f1-3 in vivo, revealing distinct roles in dividing versus differentiating cells and in normal versus cancer-like cell cycles. PMID- 20016606 TI - Optical gain in DNA-DCM for lasing in photonic materials. AB - We present a detailed study of the gain length in an active medium obtained by doping of DNA strands with 4-(dicyanomethylene)-2-methyl-6-(4 dimethylaminostyryl)-4H-pyran dye molecules. The superior thermal stability of the composite and its low quenching permit one to obtain an optical gain coefficient larger than 300 cm(-1). We also demonstrate that such an active material is feasible for the infiltration into photonic nanostructures, allowing one to obtain fluorescent photonic crystals and promising lasing properties. PMID- 20016607 TI - Tunable-diode-laser spectroscopy of C(2)H(2) using a 3.03 microm GaInAsSb/AlGaInAsSb distributed-feedback laser. AB - The mid-IR region beyond 3 microm is very attractive for gas sensing, since the fundamental absorption bands of several hydrocarbons are located in this range. We demonstrated, for the first time to our knowledge, the use of a novel GaInAsSb/AlGaInAsSb distributed-feedback laser emitting around 3.03 microm in a tunable-diode laser-spectroscopy application. The laser operates in continuous mode at room temperature with excellent single-mode and tuning properties. A comparison of the measurement results was made with the recently updated data on C212H(2) found in the HITRAN 2008 compilation. A good agreement was found between the measurements and the database. Wavelength modulation spectroscopy of acetylene at ambient conditions was made, and a sensitivity of 18 ppb (parts per billion) per meter at an integration time of 3 s corresponding to a relative absorption of 5 x 10(-6) was obtained. The optimum detection limit of the acetylene measurement in this wavelength modulation spectroscopy setup was better than 1.5 ppb m at an integration time of 600 seconds. PMID- 20016603 TI - Mammalian SUMO E3-ligases PIAS1 and PIAS4 promote responses to DNA double-strand breaks. AB - DNA double-strand breaks (DSBs) are highly cytotoxic lesions that are generated by ionizing radiation and various DNA-damaging chemicals. Following DSB formation, cells activate the DNA-damage response (DDR) protein kinases ATM, ATR and DNA-PK (also known as PRKDC). These then trigger histone H2AX (also known as H2AFX) phosphorylation and the accumulation of proteins such as MDC1, 53BP1 (also known as TP53BP1), BRCA1, CtIP (also known as RBBP8), RNF8 and RNF168/RIDDLIN into ionizing radiation-induced foci (IRIF) that amplify DSB signalling and promote DSB repair. Attachment of small ubiquitin-related modifier (SUMO) to target proteins controls diverse cellular functions. Here, we show that SUMO1, SUMO2 and SUMO3 accumulate at DSB sites in mammalian cells, with SUMO1 and SUMO2/3 accrual requiring the E3 ligase enzymes PIAS4 and PIAS1. We also establish that PIAS1 and PIAS4 are recruited to damage sites via mechanisms requiring their SAP domains, and are needed for the productive association of 53BP1, BRCA1 and RNF168 with such regions. Furthermore, we show that PIAS1 and PIAS4 promote DSB repair and confer ionizing radiation resistance. Finally, we establish that PIAS1 and PIAS4 are required for effective ubiquitin-adduct formation mediated by RNF8, RNF168 and BRCA1 at sites of DNA damage. These findings thus identify PIAS1 and PIAS4 as components of the DDR and reveal how protein recruitment to DSB sites is controlled by coordinated SUMOylation and ubiquitylation. PMID- 20016608 TI - Improved metal-semiconductor position detector with oscillating lateral photovoltaic effect. AB - The most outstanding feature of a lateral photovoltaic effect (LPE) is its linear response to the light position. We report an oscillating LPE in a modified metal semiconductor (MS) structure, in which the lateral photovoltage (LPV) will oscillate with light position. Compared with the linear LPV in that conventional MS structure, this effect shows a controllable oscillation with sharp change with light position, opening a new method of detecting very small displacement with much higher precision and sensitivity. PMID- 20016609 TI - Optical fields of the lowest modes in a uniformly active thin subwavelength spiral microcavity. AB - A numerical study is presented of several lowest in frequency modes in a spiral microlaser. The modes in an arbitrarily shaped active cavity are considered as solutions to the two-dimensional eigenproblem for the Muller boundary-integral equations. After discretization using the Nystrom-type algorithm, the eigenvalues are found in terms of frequency and material-gain threshold. PMID- 20016610 TI - Bloch-mode extraction from near-field data in periodic waveguides. AB - We demonstrate that the spatial profiles of both propagating and evanescent Bloch modes in a periodic structure can be extracted from a single measurement of an electric field at the specified optical wavelength. We develop a systematic extraction procedure by extending the concepts of high-resolution spectral methods previously developed for temporal data series to take into account the symmetry properties of Bloch modes simultaneously at all spatial locations. We illustrate the application of our method to a photonic crystal waveguide interface and confirm its robustness in the presence of noise. PMID- 20016611 TI - Quantitative analysis of forward and backward second-harmonic images of collagen fibers using Fourier transform second-harmonic-generation microscopy. AB - Fourier transform second-harmonic generation (SHG) microscopy has been applied to quantitatively compare the information content between SHG images obtained from the forward and backward direction for three tissue types: porcine tendon, sclera, and ear cartilage. Both signal types yield consistent information on the preferred orientation of collagen fibers. For all specimens, the Fourier transform of the forward and backward SHG images produces several overlapping peaks in the magnitude spectrum at various depths into the tissues, indicating that some information present in the forward SHG images can be extracted from the backward SHG images. This study highlights the potential of backward SHG microscopy for medical diagnostics. PMID- 20016612 TI - Variable optical activation of human cone photoreceptors visualized using a short coherence light source. AB - It has been shown that after a visible stimulus, optical oscillations of nearly all cone photoreceptors can be observed using long coherence length light and in a few cones using short coherence length light. Here, we show that after exposure to a visible stimulus, a short coherence length imaging source reveals light evoked oscillation signals in a large number of cones. More than 80% of cones in a given retinal area are activated (modulation in the reflectance signal) after stimulation, and the pattern of their activation can be subjectively classified into one of four categories. The application of light-evoked signal detection techniques for in vivo retinal imaging may prove useful for assessing the functional status of cones in normal and diseased retinae. PMID- 20016613 TI - Si rib waveguide photodetector with an ordered array of Ge islands for 1.5 microm. AB - This work demonstrates a rib waveguide photodetector based on a vertical Si p-i-n junction with Ge islands operating in the spectral region around lambda=1.55 microm at room temperature. A vertical stack of four layers of Ge islands is grown by molecular beam epitaxy on a silicon-on-insulator. Each layer is organized in a two-dimensional square grid with a period of 460 nm. The spectral response of the detector extends well beyond 1.6 mum at 300 K. The absorption length of approximately 135 microm (at 1/e decrease of intensity) at lambda=1.55 microm along the waveguide allows for relatively small-size devices for all-on one-platform integration. PMID- 20016614 TI - Control of the optical Kerr effect in chirped-pulse-amplification systems using model-based phase shaping. AB - Using phase shaping, the impact of the Kerr effect in a fiber-based chirped-pulse amplification (CPA) system is experimentally controlled. The technique is based on an analytical model describing the spectral phase owing to self-phase modulation in CPA systems. The method relies neither on complex phase measurements nor on time-consuming optimization routines. Nearly transform limited pulses with energies as high as 1 mJ are produced, and a B integral being as high as 8 rad is accumulated in the main amplifier. The value of the B integral is determined by the method itself. PMID- 20016615 TI - Accelerating beams. AB - We demonstrate that any two-dimensional accelerating beam can be described in a canonical form in Fourier space. In particular, we demonstrate that there is a one-to-one correspondence between complex functions in the real line (the line spectrum) and accelerating beams. An arbitrary line spectrum can be used to generate novel accelerating beams with diverse transverse shapes. The line spectra for the special cases of the families of Airy and accelerating parabolic beams are provided. PMID- 20016616 TI - Photonic scanning receiver using an electrically tuned fiber Bragg grating. AB - A 5-cm-long electrically tuned fiber Bragg grating is used to filter a microwave signal on an optical carrier at 1.55 mum. A chirped distributed-feedback structure is employed, with a transmission bandwidth of 54 MHz and relative optical carrier rejection of >30 dB for rf frequencies >2 GHz. The rapid monotonic sweep of the Bragg wavelength is translated into a fast-frequency sweep for rf analysis. PMID- 20016617 TI - Photothermal measurement of absorption and scattering losses in thin films excited by surface plasmons. AB - We present a novel noncontact, photothermal technique, based on the focus error signal of a commercial CD pickup head that allows direct determination of absorption in thin films. Combined with extinction methods, this technique yields the scattering contribution to the losses. Surface plasmon polaritons are excited using the Kretschmann configuration in thin Au films of varying thickness. By measuring the extinction and absorption simultaneously, it is shown that dielectric constants and thickness retrieval leads to inconsistencies if the model does not account for scattering. PMID- 20016618 TI - Monolithic double-grating phase mask for large-period highly coherent grating printing. AB - A monolithic double-grating phase mask comprising three short-pitch grating sections of spatial frequencies k(1) and k(2) collocated at one side of a substrate produces a large-period interferogram without higher harmonics to print in a photoresist film a latent grating of small spatial frequency equal to twice k(2)-k(1). When incorporated in a write-on-the-fly scheme, the elements permit the fabrication of unlimitedly long gratings. PMID- 20016619 TI - Demonstration of a large-size real-time full-color three-dimensional display. AB - A large-size and full-color three-dimensional (3D) display system without the need for special eyeglasses is demonstrated. With a specially fabricated holographic functional screen with a size of 1.8x1.3 m(2), the system including optimally designed camera-projector arrays and a video server can display the fully continuous, natural 3D scene with more than 1 m image depth in real time. We explain the operating principle and present experimental results. PMID- 20016620 TI - Plasma waveguide array induced by filament interaction. AB - We demonstrate that interference-assisted coalescence of two noncollinearly overlapped filaments creates a wavelength-scale periodic plasma density modulation to guide the input pulses equivalently as a photonic crystal plasma waveguide. The periodic self-channeling is evidenced by the direct observation of the filament coalescence, which reveals wavelength-scale spatial widths and periodicity dependent on the crossing angles and intensity ratios between the incident filaments. PMID- 20016621 TI - Common-path laser interferometer. AB - A common-path laser interferometer is introduced. Its most important feature is that two optical arms are entirely in a common path. Laser Doppler and laser polarization interference technologies are used in the proposed optical system. An experiment setup is established. The primary experimental result shows that this common path method is very feasible. The resolution is 0.72 nm within a 0.01 mm range, and the highest velocity is 0.56 mm/s. There is much room to improve its performance. PMID- 20016622 TI - Simulation study of surface-plasmon-resonance electro-optic light modulator based on a polymer grating coupler. AB - An electro-optic light modulator design based on a grating-coupled surface plasmon-resonance structure is numerically investigated using finite-difference time-domain simulation. The thickness effect of a dielectric layer acting as a waveguide on metal structures is investigated. The results show that the new structure has much higher modulation index than the conventional one and can implement an electro-optic modulator with low operating voltage. PMID- 20016623 TI - Onset of the optical damage in CaF(2) optics caused by deep-UV lasers. AB - The exterior sides of calcium fluoride (CaF(2)) outcoupling mirrors are damaged by ArF laser light irradiation with high pulse-energy densities (80 mJ/cm(2)). The damage is generated by a partial alteration of the CaF(2) substrate to calcite. The CaF(2) decomposition is driven by photochemical processes due to the UV light and the presence of water vapor and is supported by elevated temperatures within the laser beam transmitting area. Small filaments act as starting points for the decomposition process, where kerogenous carbon and calcite can occur. PMID- 20016624 TI - On-chip tunable long-period grating devices based on liquid crystal photonic bandgap fibers. AB - We design and fabricate an on-chip tunable long-period grating device by integrating a liquid crystal photonic bandgap fiber on silicon structures. The transmission axis of the device can be electrically rotated in steps of 45 degrees as well as switched on and off with the response time in the millisecond range. The strength of the loss peak is controlled electrically, and the spectral position of the loss peak is thermally tunable. This compact design results in a stable grating and permits this device to be more easily applied in practical systems. PMID- 20016625 TI - Double-locked semiconductor laser for radio-over-fiber uplink transmission. AB - The nonlinear dynamics of an optically injected semiconductor laser are explored for radio-over-fiber uplink transmission. Under optical injection locking, the laser at the base station is operated in the period-one oscillation state, where its intensity oscillates at a tunable microwave frequency. When the oscillation is tuned to the subcarrier frequency, it is further locked by the uplink microwave signal. By simply using an ordinary 2.5-Gbps-grade semiconductor laser, uplink transmission of the phase-shift keying (PSK) signal at a subcarrier of 16 GHz with bit-error rate of less than 10(-11) is demonstrated experimentally. Microwave PSK to optical PSK is achieved at the double-locked laser, which allows all-optical demodulation without any high-speed microwave electronics. PMID- 20016626 TI - All-optical tunable pulse frequency chirp via slow light. AB - We theoretically investigate slow light via stimulated Raman scattering, paying special attention to the picosecond regime where chromatic dispersion and cross phase modulation must be considered. In addition to the control of the Raman pulse walk-off, we demonstrate that the cross-phase-modulation-induced frequency chirp can also be all-optically tuned via Raman slow light. We further demonstrate that this new implication is a consequence of the fact that the group velocity is significantly more affected than the phase velocity in slow-light media. PMID- 20016627 TI - Hysteresis of dissipative soliton molecules in mode-locked fiber lasers. AB - We numerically demonstrate bistability of a robust dissipative soliton molecule with independently evolving phase in a normal dispersion mode-locked fiber laser. It is shown that the separation of the peaks exhibits a hysteretic behavior as a function of the pump energy. Physically this corresponds to the coexistence of two bound-state branches, one with chaotic dynamics and another one with periodic dynamics. The crucial role of gain saturation for the appearance of the hysteresis is revealed. We observe the period-doubling route to chaos. PMID- 20016628 TI - Bend insensitive optical fiber with ultralow bending loss in the visible wavelength band. AB - We report experimental development of a single-mode optical fiber optimized for a wavelength band of 400 nm to 700 nm, which showed negligible bending loss of 0.09 dB/loop at 400 nm and 0.23 dB/loop at 633 nm for a loop diameter of 10 mm. PMID- 20016629 TI - Nanocrystalline LaOCl:Tb(3+)/Sm(3+) as promising phosphors for full-color field emission displays. AB - Nanocrystalline LaOCl:Tb(3+)/Sm(3+) phosphors were synthesized by a Pechini-type sol-gel process. Under UV and electron-beam excitation, LaOCl:Tb(3+)/Sm(3+) show the characteristic emission of Tb(3+) ((5)D(3,4)-->(7)F(6),...2) and Sm(3+) ((4)G(5/2)-->(6)H(5/2,7/2,9/2)), respectively. In particular, the cathodoluminescence (CL) color of LaOCl:Tb(3+) can be tuned from blue to green by changing Tb(3+)-doped concentration, and their CL intensities (brightness) are higher than those of commercial products Y(2)SiO(5):Ce(3+) and ZnO:Zn, respectively. White CL can be realized by codoping with Tb(3+) and Sm(3+) in a single-phase LaOCl host. The obtained white light is very close to the standard white light. These phosphors are promising for application in field-emission displays. PMID- 20016630 TI - Broadband, high-power, continuous-wave, mid-infrared source using extended phase matching bandwidth in MgO:PPLN. AB - We report a compact and viable source of broadband, high-power, cw, mid-IR radiation based on a singly resonant optical parametric oscillator (SRO) pumped by a wide-bandwidth cw Yb fiber laser centered at 1060 nm. By exploiting the extended phase-matching bandwidth in a 50 mm crystal of MgO:PPLN and a ring SRO cavity, we obtain 5.3 W of broadband idler output for 25.5 W of pump at >80% depletion, transferring a pump bandwidth of 73.9 cm(-1) to an idler spectrum spread across an equal bandwidth centered at 3454 nm. By deploying output coupling of the signal, we generate 11.2 W of total power at 44% extraction efficiency with a pump depletion of >73% at the maximum available pump power. Measurements of transverse modal power confirm Gaussian distribution of signal and idler beams. PMID- 20016631 TI - Spectral broadening of mid-infrared femtosecond pulses in GaAs. AB - We achieved efficient spectral broadening for mid-IR pulses of few-microjoule energy. The spectral bandwidth of the femtosecond pulses at the center wavelength of 5000 nm increased from 540 nm to 2060 nm (from 220 to 910 cm(-1) in frequency) by nonlinear propagation in a gallium arsenide single crystal. The spectral broadening was accompanied by nonlinear absorption loss of 25%. The demonstrated scheme should be available at any operation wavelength within the material transparency range and provides a useful tool in nonlinear vibrational spectroscopy. PMID- 20016632 TI - Picosecond optical vortex converted from multigigahertz self-mode-locked high order Hermite-Gaussian Nd:GdVO(4) lasers. AB - We report on a gigahertz self-mode-locked high-order Hermite-Gaussian (HG) Nd:GdVO(4) laser. With a pump power of 2.2 W, the average output power for the TEM(0,m) modes from m=9 to m=0 are among 350-780 mW at a repetition rate of 3.5 GHz. The mode-locked pulse width is in the range of 20-25 ps for various HG TEM(0,m) modes. With a simple cylindrical-lens converter, the mode-locked HG beams are converted to generate picosecond optical vortex pulses. PMID- 20016633 TI - Enhancement of semiconducting single-wall carbon-nanotube photoluminescence. AB - Photoluminescence properties of semiconducting single-wall carbon-nanotube (s SWNT) thin films with different metallic single-wall carbon-nanotube (m-SWNT) concentrations are reported. s-SWNT purified samples are obtained by polymer assisted selective extraction. We show that the presence of a few m-SWNTs in the sample generates a drastic quenching of the emission. Therefore, the highly purified s-SWNT film is a strongly luminescent material and a good candidate for future applications in photonics, such as near-IR emitters, modulators, and detectors. PMID- 20016634 TI - Suppression of FM-to-AM conversion in third-harmonic generation at the retracing point of a crystal. AB - FM-to-AM conversion can cause many negative effects (e.g., reducing of margin against damage to the optics) on the performance of a third-harmonic conversion system. In this Letter, the FM-to-AM conversion effect in third-harmonic generation is investigated both at and away from the retracing point of Type II potassium dihydrogen phosphate crystal. Obtained results indicate that the FM-to AM conversion can be suppressed effectively when the crystal works at the retracing point. PMID- 20016635 TI - Sub-two-cycle pulses by soliton self-compression in highly nonlinear photonic crystal fibers. AB - We demonstrate the direct generation of sub-two-cycle pulses by soliton self compression of femtosecond pulses from a Ti:sapphire laser at 85 MHz using a 4.85 mm-long highly nonlinear photonic crystal fiber. Sub-nanojoule, 41 fs input pulses were compressed down to 4.6 fs without additional phase compensation schemes. To our knowledge, these are the shortest pulses obtained by soliton effect compression of a laser oscillator. Efficient, near-dispersionless collimation of the fiber output was achieved with a simple lens and an octave spanning double-chirped mirror pair. The full electric field of the compressed pulses was retrieved with a genetic algorithm applied to spectral and interferometric autocorrelation measurements, and the results are well described by numerical simulations. PMID- 20016636 TI - Spatiotemporal amplitude and phase retrieval of space-time coupled ultrashort pulses using the Shackled-FROG technique. AB - We demonstrate the validity of the Shackled-frequency-resolved-optical-gating technique for the complete characterization, both in space and in time, of ultrashort optical pulses that present strong angular dispersion. Combining a simple imaging grating with a Hartmann-Shack sensor and standard frequency resolved-optical-gating detection at a single spatial position, we are able to retrieve the full spatiotemporal structure of a tilted pulse. PMID- 20016637 TI - Gas-phase single-shot thermometry at 1 kHz using fs-CARS spectroscopy. AB - Single-laser-shot temperature measurements at a data rate of 1 kHz employing femtosecond coherent anti-Stokes Raman scattering (fs-CARS) spectroscopy of N(2) are demonstrated. The measurements are performed using a chirped-probe pulse to map the time-dependent frequency-spread dephasing of the Raman coherence, which is created by approximately 80-fs pump and Stokes beams, into the spectrum of the coherent anti-Stokes Raman scattering signal pulse. Temperature is determined from the spectral shape of the fs-CARS signal for probe delays of approximately 2 ps with respect to the pump-Stokes excitation. The accuracy and precision of the measurements for the 300-2400 K range are found to be approximately 1%-6% and approximately 1.5%-3%, respectively. PMID- 20016638 TI - Second-harmonic generation with magnetic-field controllability. AB - Based on density functional theory with the generalized gradient approximation plus on-site Coulomb repulsion method, we study the magnetic-ordering dependence of second-harmonic generation (SHG) in a polar magnet BiCoO(3). The large second order optical susceptibility, which can reach 3.7x10(-7) esu, exhibits a strong magnetic-ordering dependence, giving rise to magnetic-field controllable SHG response in polar magnets. PMID- 20016639 TI - Compressed sensing pulse-echo mode terahertz reflectance tomography. AB - We demonstrate pulse-echo mode terahertz (THz) reflectance tomography, where scattered THz waveforms are measured using a high-resolution asynchronous-optical sampling THz time domain spectroscopy (AOS THz-TDS) technique, and 3-D tomographic reconstruction is accomplished using a compressed sensing approach. One of the main advantages of the proposed system is a significant reduction of acquisition time without sacrificing the reconstruction quality, thanks to the sufficient incoherency in the pulse-echo mode-sensing matrix and the fast sampling scheme in AOS THz-TDS. PMID- 20016640 TI - Cross-spectral purity of electromagnetic fields. AB - We extend Mandel's scalar-wave concept of cross-spectral purity to electromagnetic fields. We show that in the electromagnetic case, assumptions similar to the scalar cross-spectral purity lead to a reduction formula, analogous with the one introduced by Mandel. We also derive a condition that shows that the absolute value of the normalized zeroth two-point Stokes parameter of two cross-spectrally pure electromagnetic fields is the same for every frequency component of the field. In analogy with the scalar theory we further introduce a measure of the cross-spectral purity of two electromagnetic fields, namely, the degree of electromagnetic cross-spectral purity. PMID- 20016641 TI - Compensating loss with gain in slow-light propagation along slab waveguide with anisotropic metamaterial cladding. AB - We explore slow-light propagation in a dielectric slab waveguide with anisotropic metamaterial cladding, which could be realized through multilayer structure with alternating metal and dielectric films. We show that although the loss in the realistic metamaterial will destroy the zero-energy velocity condition, the loss effect can be fully compensated by incorporating gain in the core material, which recovers the stopping and storing of light propagation in the waveguide. We demonstrate full-wave electromagnetic simulations on the realistic waveguide that validate our theoretical analysis. PMID- 20016642 TI - Time gating of heralded single photons for atomic memories. AB - We demonstrate a method for time gating the standard heralded cw spontaneous parametric downconverted single-photon source by using pulsed pumping of an optical parametric oscillator below threshold. The narrow bandwidth, high purity, high spectral brightness, and pseudodeterministic character make the source highly suitable for light-atom interfaces with atomic memories. PMID- 20016643 TI - Dual-comb electric-field cross-correlation technique for optical arbitrary waveform characterization. AB - We present an electric-field cross-correlation technique that uses a pair of frequency combs to sweep phase and group delays independently without a mechanical stage. We demonstrate this technique for characterization of optical arbitrary waveforms composed of ~30 spectral lines from a 10 GHz frequency comb. Rapid data acquisition (tens of microseconds) enables interferometric spectral phase measurement of pulses subject to propagation over 20 km of optical fiber. PMID- 20016644 TI - Microring quadrature modulators. AB - A high-Q microring integrated with a dual-2x2-Mach-Zehnder interferometer coupler is proposed as a low-chirp, low-distortion, low-drive-power, quadrature modulator capable of M-ary quadrature amplitude modulation and phase-shift keyed outputs. The modulation rate can greatly exceed the resonator linewidth and is limited only by the coupler response or free spectral range. PMID- 20016645 TI - Waveguide-saturable absorber fabricated by femtosecond pulses in YAG:Cr(4+) crystal for Q-switched operation of Yb-fiber laser. AB - A waveguide-saturable absorber with low propagation loss is fabricated by femtosecond pulses in YAG:Cr(4+) crystal. Q-switch operation of a Yb fiber laser with the new saturable absorber having absorption saturation parameters similar to the bulk YAG:Cr(4+) crystal is demonstrated. PMID- 20016646 TI - Solid-state ring laser gyro behaving like its helium-neon counterpart at low rotation rates. AB - Nonlinear couplings induced by crystal diffusion and spatial inhomogeneities of the gain have been suppressed over a broad range of angular velocities in a solid state ring laser gyro by vibrating the gain crystal at 168 kHz and 0.4 microm along the laser cavity axis. This device behaves in the same way as a typical helium-neon ring laser gyro, with a zone of frequency lock-in (or dead band) resulting from the backscattering of light on the cavity mirrors. Furthermore, it is shown that the level of angular random-walk noise in the presence of mechanical dithering depends only on the quality of the cavity mirrors, as is the case with typical helium-neon ring laser gyros. PMID- 20016647 TI - Hopf-Ranada linked and knotted light beam solution viewed as a null electromagnetic field. AB - The Hopf-Ranada linked and knotted light beam solution, which has been interpreted physically and extended analytically by Irvine and Bouwmeester recently, is viewed in this Letter as a null electromagnetic field. It is shown, in particular, that the Hopf-Ranada solution is a variant of a luminal null electromagnetic wave due originally to Robinson and Troutman and reported by Bialynicki-Birula recently. This analogy is motivated by means of a method due to Whittaker and Bateman, and a relationship to well-known scalar luminal localized waves is examined. PMID- 20016648 TI - Side-hole fiber sensor based on surface plasmon resonance. AB - Surface plasmon resonance (SPR) is demonstrated in a microstructured optical fiber sensor for the first time (to our knowledge). SPR features were observed at 560 and 620 nm when sample fluids of refractive indices n=1.38 and n=1.41, respectively, were applied to the sensor. This study also identifies a new approach to improve the resolution of fiber SPR sensors. PMID- 20016649 TI - Experimental realization of plasmonic filters for multispectral and dual polarization optical detection. AB - We report the design and implementation of a new class of plasmonic filters that are both wavelength and polarization selective. The plasmonic filter consists of a five-layer metallic photonic crystal structure and operates inside the photonic bandgap regime. We show that by manipulating the middle layer geometry alone, it is possible to tune the Fabry-Perot resonance over a broad spectral range (lambda=1.25to1.63 microm) and in a monolithic fashion. Furthermore, we show that the resonance has a definite polarization character that is determined by the orientation of the first layer grating. The plasmonic filter may be integrated with an array of photodetectors for high-throughput spectral and polarimetric imaging applications. PMID- 20016650 TI - Fast responsive nonvolatile holographic storage in LiNbO(3) triply doped with Zr, Fe, and Mn. AB - Iron and manganese doubly doped LiNbO(3) (LN:Fe,Mn) has been suggested for nonvolatile photorefractive recording; however, its response time is still of the order of minutes. Here we present results on LiNbO(3) triply doped with zirconium, iron, and manganese (LN:Zr,Fe,Mn). The codoping with Zr eliminates undesirable intrinsic traps, which strongly enhances the charge transition speed. The response time of LN:Zr,Fe,Mn for nonvolatile holographic storage shortens to only 0.95 s (wavelength of 532 nm and intensity of 400 mW/cm(2)), and the sensitivity reaches 1.31 cm/J. Thus it seems that we have found an excellent recording medium for practical holographic storage devices. PMID- 20016652 TI - Repetition-rate-selective, wavelength-tunable mode-locked laser at up to 640 GHz. AB - We demonstrate a tunable passively mode-locked fiber laser with a selectable repetition rate of up to 640 GHz. The mode-locking mechanism is based on dissipative four-wave mixing in combination with a programmable optical processor as the spectral filter. We achieve up to 20 nm wavelength tunability and present mode-locked operation at repetition rates between 40 and 640 GHz. Measurements of the power spectra using a cross-phase modulation technique confirm the mode locking. PMID- 20016651 TI - Intravital imaging of amyloid plaques in a transgenic mouse model using optical resolution photoacoustic microscopy. AB - We report optical-resolution photoacoustic microscopy (OR-PAM) for in vivo imaging of amyloid plaques in an Alzheimer's disease mouse model. Validation using conventional fluorescence microscopy and multiphoton microscopy shows that OR-PAM has sufficient sensitivity and spatial resolution to identify amyloid plaques in living brains. In addition, with dual-wavelength OR-PAM, the three dimensional morphology of amyloid plaques and the surrounding microvasculature are imaged simultaneously through a cranial window without angiographic contrast agents. OR-PAM, capable of providing both exogenous molecular contrast and endogenous hemoglobin contrast, has the potential to serve as a new technology for in vivo microscopic observations of cerebral plaque deposits. PMID- 20016653 TI - Plasmon-enhanced total-internal-reflection fluorescence by momentum-mismatched surface nanostructures. AB - We investigate optimum plasmon-enhanced total-internal-reflection fluorescence imaging by metallic thin films and nanostructures. The enhancement is based on the mismatch between the conditions of plasmon resonance and maximal near-field intensity. We have calculated plasmon-associated near-field and far-field characteristics using rigorous coupled-wave analysis. Near-field intensity was experimentally measured with fluorescent beads on silver thin films, nanogratings, and nanoislands. The results for nanostructure-based plasmon excitation confirm that momentum mismatching when exciting plasmons can increase the consequent emission of fluorescence substantially. The improvement can be critical depending on the specific structure. PMID- 20016654 TI - Observation of nonlinear Airy-like beam evolution in lithium niobate. AB - We report the observation of Gaussian beam fragmentation into Airy-like waveforms during nonlinear propagation. The effect is supported by the high-intensity photovoltaic nonlinearity arising in unbiased pure congruent lithium niobate. The process is found to occur when the nonlinear response is dominated by the nonlocal effects associated with the charge-displacement process. PMID- 20016655 TI - Polarization conversion spectroscopy of hybrid modes. AB - Enhanced polarization conversion in reflection for the Otto and Kretschmann configurations is introduced as a new method for hybrid-mode spectroscopy. Polarization conversion in reflection occurs when hybrid modes are excited in a guiding structure composed of at least one anisotropic medium. In contrast to a dark dip, in this case modes are associated with a peak in the converted reflectance spectrum, increasing the detection sensitivity and avoiding confusion with reflection dips associated with other processes, such as transmission. PMID- 20016656 TI - Phase-modulation method for AWG phase-error measurement in the frequency domain. AB - We report a phase-modulation method for measuring arrayed waveguide grating (AWG) phase error in the frequency domain. By combining the method with a digital sampling technique that we have already reported, we can measure the phase error within an accuracy of +/-0.055 rad for the center 90% waveguides in the array even when no carrier frequencies are generated in the beat signal from the interferometer. PMID- 20016657 TI - Multiple-image encryption and multiplexing using a modified Gerchberg-Saxton algorithm and phase modulation in Fresnel-transform domain. AB - What we believe to be a new technique, based on a modified Gerchberg-Saxton algorithm (MGSA) and a phase modulation scheme in the Fresnel-transform domain, is proposed to reduce cross talks existing in multiple-image encryption and multiplexing. First, each plain image is encoded and multiplexed into a phase function by using the MGSA and a different wavelength/position parameter. Then all the created phase functions are phase modulated to result in different shift amounts of the reconstruction images before being combined together into a single phase-only function. Simulation results show that the cross talks between multiplexed images have been significantly reduced, compared with prior methods [Opt. Lett.30, 1306 (2005); J. Opt. A8, 391 (2006)], thus presenting high promise in increasing the multiplexing capacity and encrypting grayscale and color images. PMID- 20016658 TI - Flexible fabrication of three-dimensional optical-domain photonic crystals using a combination of single-laser-exposure diffractive-optics lithography and template inversion. AB - We demonstrate inversion of three-dimensional photonic crystal templates fabricated in a large area with diffractive-optics lithography. A custom-designed two-dimensional diffractive optical element was used to generate highly uniform, bicontinuous, three-dimensional photonic crystal templates in a single-laser exposure. Chemical vapor deposition successfully infiltrated the thick periodic polymer structure to deposit amorphous silica and thereby define an all-silica inverted photonic crystal after polymer removal, as confirmed by focused ion-beam milling and energy-dispersive x-ray spectroscopy. The diffractive-optics lithography permitted a large number of uniform layers to form that manifested in the recording of a strong -28 dB stopband in the telecom band. PMID- 20016659 TI - Blue-light-emitting ZnSe random laser. AB - We observed blue-light-emitting random laser action with coherent feedback in ZnSe particles. Below the threshold excitation, we observed a broad spontaneous emission band peaking at approximately 470 nm. Above the threshold, several discrete lasing lines appear at the center of the spontaneous emission band (approximately 475 nm). The linewidth of each lasing emission is less than 0.4 nm, which is about 40 times narrower than that of the spontaneous emission band. Although the lasing intensity of each line varies from pulse to pulse, the peak wavelengths do not shift significantly. The lasing emission was also found to radiate in all directions. PMID- 20016660 TI - Controllable optical demultiplexing using continuously tunable optical parametric delay at 160 Gbit/s with <0.1 ps resolution. AB - We experimentally demonstrate the use of continuously tunable optical parametric delay at 160 Gbit/s, based on conversion/dispersion for demultiplexing a 160 Gbit/s signal to 40 Gbit/s. A 15.2 ns delay is shown for a 160 Gbit/s signal, with <0.1 ps resolution, achieved by inserting cascaded acousto-optic modulators after the pump laser for delay fine tuning. PMID- 20016661 TI - Chemoselective Three-Component Coupling via A Tandem Pd Catalyzed Boron-Heck and Suzuki Reaction. AB - Reported herein is a new approach to prepare biaryl derivatives via a tandem Pd catalyzed boron-Heck and Suzuki reaction. This one-pot tandem process avoided purification or addition of extra catalyst between steps. The resulting biaryl compounds can be prepared with substrate variability in good to moderate yields. PMID- 20016662 TI - Confidence Intervals for A Common Mean with Missing Data with Applications in AIDS Study. AB - In practical data analysis, nonresponse phenomenon frequently occurs. In this paper, we propose an empirical likelihood based confidence interval for a common mean by combining the imputed data, assuming that data are missing completely at random. Simulation studies show that such confidence intervals perform well, even the missing proportion is high. Our method is applied to an analysis of a real data set from an AIDS clinic trial study. PMID- 20016663 TI - Photophysical Characterization of Imidazolium-Substituted Pd(II), In(III), and Zn(II) Porphyrins as Photosensitizers for Photodynamic Therapy. PMID- 20016664 TI - Fluorescence lifetime properties of near-infrared cyanine dyes in relation to their structures. AB - Structurally diverse near-infrared (NIR) absorbing polymethine dyes were prepared and their fluorescence lifetimes (FLT) were evaluated in relation to their structural features. Comparative FLT analysis based on the modification of methine chain length and heterocyclic system showed that indolium or benz[e]indolium heptamethine dyes exhibited longer FLT than the benz[c,d]indolium trimethine dye. Modification of heterocyclic system alone with an intact chain length showed that indolium-based heptamethine dyes showed approximately 30% longer FLT than the benz[e]indolium-based dyes. In general, the FLT of polymethine dyes increased from polar to non-polar solvents. In addition, correlation study between the theoretical and the experimental FLT for indocyanine green (ICG) suggests that the lack of structural rigidity for these cyanine dyes is primarily responsible for the loss of the excited state energy via non-radiative pathway. PMID- 20016665 TI - Using an Approximate Bayesian Bootstrap to Multiply Impute Nonignorable Missing Data. AB - An Approximate Bayesian Bootstrap (ABB) offers advantages in incorporating appropriate uncertainty when imputing missing data, but most implementations of the ABB have lacked the ability to handle nonignorable missing data where the probability of missingness depends on unobserved values. This paper outlines a strategy for using an ABB to multiply impute nonignorable missing data. The method allows the user to draw inferences and perform sensitivity analyses when the missing data mechanism cannot automatically be assumed to be ignorable. Results from imputing missing values in a longitudinal depression treatment trial as well as a simulation study are presented to demonstrate the method's performance. We show that a procedure that uses a different type of ABB for each imputed data set accounts for appropriate uncertainty and provides nominal coverage. PMID- 20016666 TI - Quantitative ToF-SIMS Studies of Protein Drug Release from Biodegradable Polymer Drug Delivery Membranes. AB - Biodegradable polymers are of interest in developing strategies to control protein drug delivery. The protein that was used in this study is Keratinocyte Growth Factor (KGF) which is a protein involved in the re-epithelialization process. The protein is stabilized in the biodegradable polymer matrix during formulation and over the course of polymer degradation with the use of an ionic surfactant Aerosol-OT (AOT) which will encapsulate the protein in an aqueous environment. The release kinetics of the protein from the surface of these materials requires precise timing which is a crucial factor in the efficacy of this drug delivery system.Time of Flight Secondary Ion Mass Spectrometry (ToF SIMS) was used in the same capacity to identify the molecular ion peak of the surfactant and polymer and use this to determine surface concentration. In the polymer matrix, the surfactant molecular ion peak was observed in the positive and negative mode at m/z 467 and 421, respectively. These peaks were determined to be [AOT + Na+] and [AOT-Na+]-. These methods are used to identify the surfactant and protein from the polymer matrix and are used to measure the rate of surface accumulation. The second step was to compare this accumulation rate with the release rate of the protein into an aqueous solution during the degradation of the biodegradable film. This rate is compared to that from fluorescence spectroscopy measurements using the protein autofluorescence from that released into aqueous solution. PMID- 20016667 TI - Improving the performance of predictive process modeling for large datasets. AB - Advances in Geographical Information Systems (GIS) and Global Positioning Systems (GPS) enable accurate geocoding of locations where scientific data are collected. This has encouraged collection of large spatial datasets in many fields and has generated considerable interest in statistical modeling for location-referenced spatial data. The setting where the number of locations yielding observations is too large to fit the desired hierarchical spatial random effects models using Markov chain Monte Carlo methods is considered. This problem is exacerbated in spatial-temporal and multivariate settings where many observations occur at each location. The recently proposed predictive process, motivated by kriging ideas, aims to maintain the richness of desired hierarchical spatial modeling specifications in the presence of large datasets. A shortcoming of the original formulation of the predictive process is that it induces a positive bias in the non-spatial error term of the models. A modified predictive process is proposed to address this problem. The predictive process approach is knot-based leading to questions regarding knot design. An algorithm is designed to achieve approximately optimal spatial placement of knots. Detailed illustrations of the modified predictive process using multivariate spatial regression with both a simulated and a real dataset are offered. PMID- 20016668 TI - Growth and Survival of Acid-Resistant and Non-Acid-Resistant Shiga-Toxin Producing Escherichia coli Strains during the Manufacture and Ripening of Camembert Cheese. AB - Growth and survival of acid-resistant (AR) and non-acid-resistant (NAR) Shiga toxin-producing Escherichia coli (STEC) strains were investigated during the manufacture and ripening of microfiltered milk Camembert cheeses. The induction of acid resistance of the STEC strains in cheeses was also studied. Six different mixtures of AR and/or NAR STEC strains were inoculated separately into microfiltered milk at a level of 10(3) CFU mL(-1). The STEC counts (AR and NAR) initially increased by 1 to 2 log(10) CFU g(-1) during cheese-making. Thereafter, the populations stabilized during salting/drying and then decreased during the early stages of ripening. Exposing the STEC strains in artificially inoculated cheeses to simulated gastric fluid (SGF - pH: 2.0) reduced the number of NAR strains to undetectable levels within 40 minutes, versus 120 minutes for the AR STEC strains. AR and NAR STEC were able to survive during the manufacture and ripening of Camembert cheese prepared from microfiltered milk with no evidence of induced acid tolerance in NAR STEC strains. PMID- 20016669 TI - Characteristics of Community-Associated Methicillin-Resistant Staphylococcus aureus (CA-MRSA) Strains Isolated from Skin and Soft-Tissue Infections in Uruguay. AB - We analyzed 90 nonduplicates community-associated methicillin-resistant S. aureus (CA-MRSA) strains isolated from skin and soft-tissue infections. All strains were mecA positive. Twenty-four of the 90 strains showed inducible macrolide lincosamide-streptogramin B resistance. All strains produced alpha-toxin; 96% and 100% of them displayed positive results for lukS-F and cna genes, respectively. Eigthy-five strains expressed capsular polysaccharide serotype 8. Six different pulsotypes were discriminated by pulsed-field gel electrophoresis (PFGE) and three predominant groups of CA-MRSA strains (1, 2, and 4) were identified, in agreement with phenotypic and genotypic characteristics. Strains of group 1 (pulsotype A, CP8+, and Panton-Valentine leukocidin (PVL)+) were the most frequently recovered and exhibited a PFGE band pattern identical to other CA-MRSA strains previously isolated in Uruguay and Brazil. Three years after the first local CA-MRSA report, these strains are still producing skin and soft-tissue infections demonstrating the stability over time of this community-associated emerging pathogen. PMID- 20016670 TI - Investigations on the effects of dietary essential oils and different husbandry conditions on the gut ecology in piglets after weaning. AB - Essential oils (EO) are being considered as possible alternatives to in-feed antibiotic growth promoters in pig nutrition. The effects of an EO mixture consisting of limonene, eugenol and pinene (10.0, 2.0, and 4.8 mg/kg diet, resp.) on gut physiology and ecology were studied in piglets. The experiment was conducted at low (commercial farm) and high hygienic conditions (experimental farm), to elucidate interactions between EO supplementation and husbandry methods. Piglets were weaned at 28 days of age, when they were offered either a control diet (C) or C with EO. Four piglets were sacrificed in each group on day 29, 30, 33 and 39. Digesta from the third distal part of the small intestine and from the colon were sampled and analysed for pH, dry matter, lactic acid, short chain fatty acids and ammonia concentrations. Enterobacteria, enterococci, lactobacilli and yeast counts were obtained by plating. Genomic DNA was extracted from digesta and polymerase chain reaction-denaturing gradient gel electrophoresis was performed. Individual microbial communities were identified at each farm. Age affected the intestinal parameters. No effects of the EO with exception for a significant reduction in colon bacterial diversity at 39 days of age could be recorded at experimental farm. PMID- 20016672 TI - The Differential Gene Expression Pattern of Mycobacterium tuberculosis in Response to Capreomycin and PA-824 versus First-Line TB Drugs Reveals Stress- and PE/PPE-Related Drug Targets. AB - Tuberculosis is a leading infectious disease causing millions of deaths each year. How to eradicate mycobacterial persistence has become a central research focus for developing next-generation TB drugs. Yet, the knowledge in this area is fundamentally limited and only a few drugs, notably capreomycin and PA-824, have been shown to be active against non-replicating persistent TB bacilli. In this study, we performed a new bioinformatics analysis on microarray-based gene expression data obtained from the public domain to explore genes that were differentially induced by drugs between the group of capreomycin and PA-824 and the group of mainly the first-line TB drugs. Our study has identified 42 genes specifically induced by capreomycin and PA-824. Many of these genes are related to stress responses. In terms of the distribution of identified genes in a specific category relative to the whole genome, only the categories of PE/PPE and conserved hypotheticals have statistical significance. Six among the 42 genes identified in this study are on the list of the top 100 persistence targets selected by the TB Structural Genomics Consortium. Further biological elucidation of their roles in mycobacterial persistence is warranted. PMID- 20016671 TI - Epithelial Cell Gene Expression Induced by Intracellular Staphylococcus aureus. AB - HEp-2 cell monolayers were cocultured with intracellular Staphylococcus aureus, and changes in gene expression were profiled using DNA microarrays. Intracellular S. aureus affected genes involved in cellular stress responses, signal transduction, inflammation, apoptosis, fibrosis, and cholesterol biosynthesis. Transcription of stress response and signal transduction-related genes including atf3, sgk, map2k1, map2k3, arhb, and arhe was increased. In addition, elevated transcription of proinflammatory genes was observed for tnfa, il1b, il6, il8, cxcl1, ccl20, cox2, and pai1. Genes involved in proapoptosis and fibrosis were also affected at transcriptional level by intracellular S. aureus. Notably, intracellular S. aureus induced strong transcriptional down-regulation of several cholesterol biosynthesis genes. These results suggest that epithelial cells respond to intracellular S. aureus by inducing genes affecting immunity and in repairing damage caused by the organism, and are consistent with the possibility that the organism exploits an intracellular environment to subvert host immunity and promote colonization. PMID- 20016673 TI - Comprehensive and rapid real-time PCR analysis of 21 foodborne outbreaks. AB - A set of four duplex SYBR Green I PCR (SG-PCR) assay combined with DNA extraction using QIAamp DNA Stool Mini kit was evaluated for the detection of foodborne bacteria from 21 foodborne outbreaks. The causative pathogens were detected in almost all cases in 2 hours or less. The first run was for the detection of 8 main foodborne pathogens in 5 stool specimens within 2 hours and the second run was for the detection of other unusual suspect pathogens within a further 45 minutes. After 2 to 4 days, the causative agents were isolated and identified. The results proved that for comprehensive and rapid molecular diagnosis in foodborne outbreaks, Duplex SG-PCR assay is not only very useful, but is also economically viable for one-step differentiation of causative pathogens in fecal specimens obtained from symptomatic patients. This then allows for effective diagnosis and management of foodborne outbreaks. PMID- 20016674 TI - Molecular Typing of Neisseria gonorrhoeae Isolates by Opa-Typing and Ribotyping in New Delhi, India. AB - Control and preventive measures for gonococcal infections are based on precise epidemiological characteristics of N. gonorrhoeae isolates. In the present study the potential utility of opa-typing and ribotyping for molecular epidemiological study of consecutive gonococcal strains was determined. Sixty gonococcal isolates were subjected to ribotyping with two restriction enzymes, AvaII and HincII, and opa-typing with TaqI and HpaII for epidemiological characterization of gonococcal population. Ribotyping with AvaII yielded 6 ribotype patterns while twelve RFLP patterns were observed with HincII. Opa-typing of the 60 isolates revealed a total 54 opa-types, which 48 were unique and 6 formed clusters. Fifty-two opa types were observed with TaqI-digested PCR product while opa-typing with HpaII demonstrated 54 opa-types. The opa-types from isolates that were epidemiologically unrelated were distinct, whereas those from the sexual contacts were identical. The results showed that opa-typing is highly useful for characterizing gonococcal strains from sexual contacts and has more discriminatory than ribotyping that could differentiate between gonococci of the same ribotype. The technique even with a single restriction enzyme has a high level of discrimination (99.9%) between epidemiologically unrelated isolates. In conclusion, the molecular methods such as opa-typing and ribotyping can be used for epidemiological characterization of gonococcal strains. PMID- 20016675 TI - Soybean Lectin Enhances Biofilm Formation by Bradyrhizobium japonicum in the Absence of Plants. AB - Soybean lectin (SBL) purified from soybean seeds by affinity chromatography strongly bound to Bradyrhizobium japonicum USDA 110 cell surface. This lectin enhanced biofilm formation by B. japonicum in a concentration-dependent manner. Presence of galactose during biofilm formation had different effects in the presence or absence of SBL. Biofilms were completely inhibited in the presence of both SBL and galactose, while in the absence of SBL, galactose was less inhibitory. SBL was very stable, since its agglutinating activity of B. japonicum cells as well as of human group A+ erythrocytes was resistant to preincubation for one week at 60 degrees C. Hence, we propose that plant remnants might constitute a source of this lectin, which might remain active in soil and thus favor B. japonicum biofilm formation in the interval between soybean crop seasons. PMID- 20016676 TI - Production of a Blue Pigment (Glaukothalin) by Marine Rheinheimera spp. AB - Two gamma-Proteobacteria strains, that is, HP1 and HP9, which both produce a diffusible deep blue pigment, were isolated from the German Wadden Sea and from the Oresund, Denmark, respectively. Both strains affiliate with the genus Rheinheimera. Small amounts of the pigment could be extracted from HP1 grown in a 50 L fermenter and were purified chromatographically. Chemical analysis of the pigment including NMR and mass spectrometry led to a molecular formula of C(34)H(56)N(4)O(4) (m.w. 584.85) which has not yet been reported in literature. The molecule is highly symmetrically and consists of two heterocyclic halves to which aliphatic side chains are attached. The pigment has been named glaukothalin due to its blue color and its marine origin (glaukos, gr. = blue, thalatta, gr. = sea). Production of glaukothalin on MB2216 agar plates by our Rheinheimera strains is affected in the presence of other bacterial strains either increasing or decreasing pigment production. The addition of a single amino acid, arginine (5 gl(-1)), greatly increases pigment production by our Rheinheimera strains. Even though the production of glaukothalin leads to inhibitory activity against three bacterial strains from marine particles, our Rheinheimera isolates are inhibited by various bacteria of different phylogenetic groups. The ecological role of glaukothalin production by Rheinheimera strains, however, remains largely unknown. PMID- 20016677 TI - Lactobacillus rhamnosus GG Suppresses Meningitic E. coli K1 Penetration across Human Intestinal Epithelial Cells In Vitro and Protects Neonatal Rats against Experimental Hematogenous Meningitis. AB - The purpose of this study was to examine prophylactic efficacy of probiotics in neonatal sepsis and meningitis caused by E. coli K1. The potential inhibitory effect of Lactobacillus rhamnosus GG (LGG) on meningitic E. coli K1 infection was examined by using (i) in vitro inhibition assays with E44 (a CSF isolate from a newborn baby with E. coli meningitis), and (ii) the neonatal rat model of E. coli sepsis and meningitis. The in vitro studies demonstrated that LGG blocked E44 adhesion, invasion, and transcytosis in a dose-dependent manner. A significant reduction in the levels of pathogen colonization, E. coli bacteremia, and meningitis was observed in the LGG-treated neonatal rats, as assessed by viable cultures, compared to the levels in the control group. In conclusion, probiotic LGG strongly suppresses meningitic E. coli pathogens in vitro and in vivo. The results support the use of probiotic strains such as LGG for prophylaxis of neonatal sepsis and meningitis. PMID- 20016678 TI - Enterobiasis in ectopic locations mimicking tumor-like lesions. AB - Both the clinical and the histopathological diagnostic difficulties of oxyuriasis in unusual sites and their importance from a clinical point of view are pointed out. The authors report two ectoptic cases of enterobiasis observed in Northern Italy, one located in a fallopian tube of a 57-year-old woman and the other in a perianal subcutaneous tissue of a 59-year-old man, mimicking tumor-like lesions. The authors take advantage of the occasion to focus the attention of the medical world on this subject, lamenting the scarce importance given to this parasitosis in university courses of medical schools and in medical textbooks as it is incorrectly considered "out-of-fashion." PMID- 20016679 TI - Shortened Time to Identify Staphylococcus Species from Blood Cultures and Methicillin Resistance Testing Using CHROMAgar. AB - The ability to rapidly differentiate coagulase-negative staphylococcus (CoNS) from Staphylococcus aureus and to determine methicillin resistance is important as it affects the decision to treat empiric antibiotic selection. The objective of this study was to evaluate CHROMagar S. aureus and CHROMagar MRSA (Becton Dickinson) for rapid identification of Staphylococcus spp. directly from blood cultures. Consecutive blood culture bottles (BacT Alert 3D SA and SN, bioMerieux) growing gram-positive cocci in clusters were evaluated. An aliquot was plated onto CHROMagar MRSA (C-MRSA) and CHROMagar S. aureus (C-SA) plates, which were read at 12 to 16 hours. C-SA correctly identified 147/147 S. aureus (100% sensitivity); 2 CoNS were misidentified as S. aureus (98% specificity). C-MRSA correctly identified 74/77 MRSA (96% sensitivity). None of the MSSA isolates grew on C-MRSA (100% specificity). In conclusion, CHROMagar is a rapid and sensitive method to distinguish MRSA, MSSA, and coagulase-negative Staphylococcus and may decrease time of reporting positive results. PMID- 20016680 TI - Simvastatin does not affect vitamin d status, but low vitamin d levels are associated with dyslipidemia: results from a randomised, controlled trial. AB - Objectives. Statin drugs act as inhibitors of the 3-hydroxy-3methylglutaryl coenzyme A (HMG-CoA) reductase enzyme early in the mevalonate pathway, thereby reducing the endogenous cholesterol synthesis. In recent studies, it has been suggested from epidemiological data that statins also may improve vitamin D status, as measured by increased plasma 25-hydroxyvitamin D (25OHD) levels. We now report the results from a randomised controlled trial on effects of simvastatin on plasma 25OHD levels. Design and Methods. We randomised 82 healthy postmenopausal women to one year of treatment with either simvastatin 40 mg/d or placebo and performed measurement at baseline and after 26 and 52 weeks of treatment. The study was completed by 77 subjects. Results. Compared with placebo, plasma levels of cholesterol and low-density lipoproteins decreased in response to treatment with simvastatin, but our study showed no effect of simvastatin on vitamin D status. However, plasma levels of triglycerides were inversely associated with tertiles of plasma 25OHD levels and changes in plasma triglycerides levels correlated inversely with seasonal changes in vitamin D status. Conclusion. Our data do not support a pharmacological effect of statins on vitamin D status, but do suggest that vitamin D may influence plasma lipid profile and thus be of importance to cardiovascular health. PMID- 20016681 TI - Vitamin d and chronic pain in immigrant and ethnic minority patients investigation of the relationship and comparison with native Western populations. AB - Vitamin D deficiency has been implicated in chronic pain. Immigrant and ethnic minority populations have been shown to have lower vitamin D levels than native Western populations and often to be vitamin D deficient. This systematic review investigates the relationship between vitamin D and chronic pain in immigrant and ethnic minority populations. Included were studies reporting on 25-OH vitamin D levels in immigrant/ethnic minority populations affected by chronic pain, and/or reporting on the treatment of chronic pain with vitamin D preparations in such populations. We found that 25-OH vitamin D levels were low and often deficient in immigrant/ethnic minority populations. Vitamin D levels depended on the latitude of the study location and hence sunlight exposure. There was insufficient evidence to reach a verdict on the value of treating chronic pain in immigrant/ethnic minority patients with vitamin D preparations because the studies were few, small, and of low quality. PMID- 20016682 TI - Vitamin d levels and lipid response to atorvastatin. AB - Adequate vitamin D levels are necessary for good vascular health. 1,25 dihydroxycholecalciferol activates CYP3A4, an enzyme of the cytochrome P450 system, which metabolizes atorvastatin to its main metabolites. The objective of this study was to evaluate the response of cholesterol and triglycerides to atorvastatin according to vitamin D levels. Sixty-three patients with acute myocardial infarction treated with low and high doses of atorvastatin were included. Levels of total cholesterol, triglycerides, HDL cholesterol, and LDL cholesterol were measured at baseline and at 12 months of follow-up. Baseline levels of 25-hydroxyvitamin D (25-OHD) were classified as deficient (<30 nmol/L), insufficient (30-50 nmol/L), and normal (>50 nmol/L). In patients with 25-OHD <30 nmol/L, there were no significant changes in levels of total cholesterol (173 +/- 47 mg/dL versus 164 +/- 51 mg/dL), triglycerides (151 +/- 49 mg/dL versus 177 +/- 94 mg/dL), and LDL cholesterol (111 +/- 48 mg/dL versus 92 45 +/- mg/dL); whereas patients with insufficient (30-50 nmol/L) and normal vitamin D (>50 nmol/L) had a good response to atorvastatin. We suggest that vitamin D concentrations >30 nmol/L may be required for atorvastatin to reduce lipid levels in patients with acute myocardial infarction. PMID- 20016683 TI - The Prevalence of Vitamin D Deficiency Is Similar between Thyroid Nodule and Thyroid Cancer Patients. AB - Introduction. There are reported associations between vitamin D deficiency and breast, prostate, and colon cancer, but the relationship in thyroid cancer has not been evaluated. Methods. We evaluated serum calcium, creatinine, albumin, and 25-hydroxy vitamin D (25-OH-D) in 42 thyroid nodule, 45 thyroid cancer in remission, and 24 active thyroid cancer patients. Results. 25-OH-D was not different between groups. The percent with 25-OH-D levels <75 nmol/L was not significantly different between groups and was not affected by season of measurement, age, or cancer stage. Multivariate regression showed a BMI of >/=30 kg/m(2) to be the only significant predictor of vitamin D deficiency. Conclusions. Rates of vitamin D deficiency are similar in thyroid nodules and thyroid cancer, although higher than the general population. This is different than previous studies for other cancers, which show higher rates of vitamin D deficiency. BMI was the only predictor of vitamin D deficiency. PMID- 20016684 TI - Structural basis for the aminoacid composition of proteins from halophilic archea. AB - Proteins from halophilic organisms, which live in extreme saline conditions, have evolved to remain folded at very high ionic strengths. The surfaces of halophilic proteins show a biased amino acid composition with a high prevalence of aspartic and glutamic acids, a low frequency of lysine, and a high occurrence of amino acids with a low hydrophobic character. Using extensive mutational studies on the protein surfaces, we show that it is possible to decrease the salt dependence of a typical halophilic protein to the level of a mesophilic form and engineer a protein from a mesophilic organism into an obligate halophilic form. NMR studies demonstrate complete preservation of the three-dimensional structure of extreme mutants and confirm that salt dependency is conferred exclusively by surface residues. In spite of the statistically established fact that most halophilic proteins are strongly acidic, analysis of a very large number of mutants showed that the effect of salt on protein stability is largely independent of the total protein charge. Conversely, we quantitatively demonstrate that halophilicity is directly related to a decrease in the accessible surface area. PMID- 20016685 TI - ZBED6, a novel transcription factor derived from a domesticated DNA transposon regulates IGF2 expression and muscle growth. AB - A single nucleotide substitution in intron 3 of IGF2 in pigs abrogates a binding site for a repressor and leads to a 3-fold up-regulation of IGF2 in skeletal muscle. The mutation has major effects on muscle growth, size of the heart, and fat deposition. Here, we have identified the repressor and find that the protein, named ZBED6, is previously unknown, specific for placental mammals, and derived from an exapted DNA transposon. Silencing of Zbed6 in mouse C2C12 myoblasts affected Igf2 expression, cell proliferation, wound healing, and myotube formation. Chromatin immunoprecipitation (ChIP) sequencing using C2C12 cells identified about 2,500 ZBED6 binding sites in the genome, and the deduced consensus motif gave a perfect match with the established binding site in Igf2. Genes associated with ZBED6 binding sites showed a highly significant enrichment for certain Gene Ontology classifications, including development and transcriptional regulation. The phenotypic effects in mutant pigs and ZBED6 silenced C2C12 myoblasts, the extreme sequence conservation, its nucleolar localization, the broad tissue distribution, and the many target genes with essential biological functions suggest that ZBED6 is an important transcription factor in placental mammals, affecting development, cell proliferation, and growth. PMID- 20016686 TI - Influenza in Africa. PMID- 20016687 TI - Ionic and pH effects on the osmotic properties and structure of polyelectrolyte gels. AB - We investigate the effects of salt concentration and pH on neutralized poly(acrylic acid) (PAA) gels in near physiological salt solutions. Either adding calcium ions or decreasing the pH are found to induce reversible volume transitions but the nature of these transitions seems to be different. For example, the osmotic pressure exhibits a simple power law dependence on the concentration as the transition is approached in both systems, but the power law exponent n is substantially different in the two cases. On decreasing the pH the value of n gradually increases from 2.1 (at pH = 7) to 3.2 (at pH = 1). By contrast, n decreases with increasing calcium ion concentration from 2.1 (in 100 mM NaCl solution) to 1.6 (0.8 mM CaCl(2) in 100 mM NaCl solution). In both systems, a strong increase of the small-angle neutron scattering intensity (SANS) is observed near the volume transition. The SANS results reveal that calcium ions favor the formation of linearly aligned regions in PAA gels. PMID- 20016688 TI - School playground surfacing and arm fractures in children: a cluster randomized trial comparing sand to wood chip surfaces. AB - BACKGROUND: The risk of playground injuries, especially fractures, is prevalent in children, and can result in emergency room treatment and hospital admissions. Fall height and surface area are major determinants of playground fall injury risk. The primary objective was to determine if there was a difference in playground upper extremity fracture rates in school playgrounds with wood fibre surfacing versus granite sand surfacing. Secondary objectives were to determine if there were differences in overall playground injury rates or in head injury rates in school playgrounds with wood fibre surfacing compared to school playgrounds with granite sand surfacing. METHODS AND FINDINGS: The cluster randomized trial comprised 37 elementary schools in the Toronto District School Board in Toronto, Canada with a total of 15,074 students. Each school received qualified funding for installation of new playground equipment and surfacing. The risk of arm fracture from playground falls onto granitic sand versus onto engineered wood fibre surfaces was compared, with an outcome measure of estimated arm fracture rate per 100,000 student-months. Schools were randomly assigned by computer generated list to receive either a granitic sand or an engineered wood fibre playground surface (Fibar), and were not blinded. Schools were visited to ascertain details of the playground and surface actually installed and to observe the exposure to play and to periodically monitor the depth of the surfacing material. Injury data, including details of circumstance and diagnosis, were collected at each school by a prospective surveillance system with confirmation of injury details through a validated telephone interview with parents and also through collection (with consent) of medical reports regarding treated injuries. All schools were recruited together at the beginning of the trial, which is now closed after 2.5 years of injury data collection. Compliant schools included 12 schools randomized to Fibar that installed Fibar and seven schools randomized to sand that installed sand. Noncompliant schools were added to the analysis to complete a cohort type analysis by treatment received (two schools that were randomized to Fibar but installed sand and seven schools that were randomized to sand but installed Fibar). Among compliant schools, an arm fracture rate of 1.9 (95% confidence interval [CI] 0.04-6.9) per 100,000 student-months was observed for falls into sand, compared with an arm fracture rate of 9.4 (95% CI 3.7-21.4) for falls onto Fibar surfaces (p< or =0.04905). Among all schools, the arm fracture rate was 4.5 (95% CI 0.26-15.9) per 100,000 student-months for falls into sand compared with 12.9 (95% CI 5.1-30.1) for falls onto Fibar surfaces. No serious head injuries and no fatalities were observed in either group. CONCLUSIONS: Granitic sand playground surfaces reduce the risk of arm fractures from playground falls when compared with engineered wood fibre surfaces. Upgrading playground surfacing standards to reflect this information will prevent arm fractures. TRIAL REGISTRATION: Current Controlled Trials ISRCTN02647424. PMID- 20016689 TI - A "one health" approach to address emerging zoonoses: the HALI project in Tanzania. PMID- 20016691 TI - Introduction of oral vitamin D supplementation and the rise of the allergy pandemic. AB - The history of the allergy pandemic is well documented, enabling us to put the vitamin D hypothesis into its historical context. The purpose of this study is to compare the prevalence of rickets, vitamin D supply, and allergy prevalence at 50 year intervals by means of a retrospective analysis of the literature since 1880. English cities in 1880 were characterized by an extremely high rickets prevalence, the beginning of commercial cod liver oil production, and the near absence of any allergic diseases. By 1930 hay fever prevalence had risen to about 3% in English-speaking countries where cod liver oil was preferentially used for the treatment of rickets. In 1980 vitamin D was used nation-wide in all industrialized countries as supplement to industrial baby food, thus eradicating nearly all cases of rickets. At the same time the allergy prevalence reached an all-time high, affecting about 30% of the population. Time trends are therefore compatible with the vitamin D hypothesis although direct conclusions cannot be drawn. It is interesting, however, to note that there are at least two earlier research papers linking synthesized vitamin D intake and allergy (Reed 1930 and Selye 1962) published prior to the modern vitamin D hypothesis first proposed in 1999. PMID- 20016690 TI - Imitators of exercise-induced bronchoconstriction. AB - Exercise-induced bronchoconstriction (EIB) is described by transient narrowing of the airways after exercise. It occurs in approximately 10% of the general population, while athletes may show a higher prevalence, especially in cold weather and ice rink athletes. Diagnosis of EIB is often made on the basis of self-reported symptoms without objective lung function tests, however, the presence of EIB can not be accurately determined on the basis of symptoms and may be under-, over-, or misdiagnosed. The goal of this review is to describe other clinical entities that mimic asthma or EIB symptoms and can be confused with EIB. PMID- 20016692 TI - Treating rhinitis in the older population: special considerations. AB - Rhinitis in the elderly is a common but often neglected condition. Structural changes in the nose associated with aging, predisposes the elderly to rhinitis. There are a number of specific factors that affect medical treatment of the elderly including polypharmacy, cognitive dysfunction, changes in body composition, impairment of liver and renal function and the cost of medications in the face of limited resources. Rhinitis in the elderly can be placed in several categories and treatment should be appropriate for each condition. The most important aim is to moisten the nasal mucosa since the nose of the elderly is so dry. Great caution should be used in treatment with first generation antihistamines and decongestants. Medications generally well tolerated by the elderly are second generation antihistamines, intra-nasal anti-inflammatory agents, leukotriene modifiers and iprapropium nasal spray. PMID- 20016693 TI - Platelet release of Vascular Endothelial Growth Factor (VEGF) in patients undergoing chemotherapy for breast cancer. AB - BACKGROUND: Venous thromboembolism (VTE) following breast cancer chemotherapy is common. Chemotherapy-induced alterations in markers of haemostasis occur during chemotherapy. In this study we investigated the changes in serum and plasma VEGF, together with platelet release of VEGF and related these to the development of VTE at 3 months. METHODS: Serum and plasma VEGF, together with platelet release of VEGF were measured prior to chemotherapy and at 24 hours; four-, eight days and three months following commencement of chemotherapy in early and advanced breast cancer patients and in age and sex matched controls. Duplex ultrasound imaging was performed after one month or if symptomatic. RESULTS: Of 123 patients 9.8% developed VTE within three months. Serum and plasma VEGF were increased in advanced breast cancer as was platelet release of VEGF. Prior to chemotherapy a 100 microg/ml increase in serum VEGF was associated with a 40% increased risk of VTE, while a 10 microg/ml increase in plasma VEGF was associated with a 20% increased risk of VTE. Serum VEGF showed a different response to chemotherapy in those who developed VTE. CONCLUSION: A group of patients at risk of VTE could be identified, allowing targeted thromboprophylaxis. Whether or not the response in VEGF during chemotherapy has any angiogenic significance remains to be elucidated. PMID- 20016694 TI - Novel insights into the differential functions of Notch ligands in vascular formation. AB - The Notch signaling pathway is a critical component of vascular formation and morphogenesis in both development and disease. Compelling evidence indicates that Notch signaling is required for the induction of arterial-cell fate during development and for the selection of endothelial tip and stalk cells during sprouting angiogenesis. In mammals, two of the four Notch receptors (Notch1 and Notch4) and three of the five Notch ligands (Jagged1, Dll1, and Dll4) are predominantly expressed in vascular endothelial cells and are important for many aspects of vascular biology. During arterial cell-fate selection and angiogenesis, the roles of Notch1 and Notch4 are thought to be similar, and the function of Dll4 is well-characterized. However, the molecular mechanisms that determine the functional similarities and differences of Notch ligands in vascular endothelial cells remain largely unknown; consequently, additional research is needed to elucidate the ligand-specific functions and mechanisms associated with Notch activation in the vascular endothelium. Results from recent studies indicate that Dll1 and Dll4 have distinct roles in the specification and maintenance of arterial cell identity, while Dll4 and Jagged1 have opposing functions in tip- and stalk-cell selection during sprouting angiogenesis. This review will focus on the newly discovered, distinct functions of several Notch ligands in the regulation of blood vessel formation and will provide perspectives for future research in the field. PMID- 20016695 TI - Blockade of vascular angiogenesis by Aspergillus usamii var. shirousamii transformed Angelicae Gigantis Radix and Zizyphus jujuba. AB - The matrix metalloproteinases (MMP) play an important role in tumor invasion, angiogenesis and inflammatory tissue destruction. Increased expression of MMP was observed in benign tissue hyperplasia and in atherosclerotic lesions. Invasive cancer cells utilize MMP to degrade the extracellular matrix and vascular basement membrane during metastasis, where MMP-2 has been implicated in the development and dissemination of malignancies. The present study attempted to examine the antiangiogenic activity of the medicinal herbs of Aspergillus usamii var. shirousamii-transformed Angelicae Gigantis Radix and Zizyphus jujube (tAgR and tZj) with respect to MMP-2 production and endothelial motility in phorbol 12 myristate 13-acetate (PMA)- or VEGF-exposed human umbilical vein endothelial cells (HUVEC). Nontoxic tAgR and tZj substantially suppressed PMA-induced MMP-2 secretion. In addition, 25 microg/mL tAgR and tZj prevented vascular endothelial growth factor-stimulated endothelial cell transmigration and tube formation. The results reveal that tAgR and tZj dampened endothelial MMP-2 production leading to endothelial transmigration and tube formation. tAgR and tZj-mediated inhibition of endothelial MMP may boost a therapeutic efficacy during vascular angiogenesis. PMID- 20016696 TI - Intestinal absorption of aloin, aloe-emodin, and aloesin; A comparative study using two in vitro absorption models. AB - Aloe products are one of the top selling health-functional foods in Korea, however the adequate level of intake to achieve desirable effects are not well understood. The objective of this study was to determine the intestinal uptake and metabolism of physiologically active aloe components using in vitro intestinal absorption model. The Caco-2 cell monolayer and the everted gut sac were incubated with 5-50 microM of aloin, aloe-emodin, and aloesin. The basolateral appearance of test compounds and their glucuronosyl or sulfated forms were quantified using HPLC. The % absorption of aloin, aloe-emodin, and aloesin was ranged from 5.51% to 6.60%, 6.60% to 11.32%, and 7.61% to 13.64%, respectively. Up to 18.15%, 18.18%, and 38.86% of aloin, aloe-emodin, and aloesin, respectively, was absorbed as glucuronidated or sulfated form. These results suggest that a significant amount is transformed during absorption. The absorption rate of test compounds except aloesin was similar in two models; more aloesin was absorbed in the everted gut sac than in the Caco-2 monolayer. These results provide information to establish adequate intake level of aloe supplements to maintain effective plasma level. PMID- 20016697 TI - Effect of Chlorella intake on Cadmium metabolism in rats. AB - This study was performed to investigate the effect of chlorella on cadmium (Cd) toxicity in Cd- administered rats. Sixty male Sprague-Dawley rats (14 week-old) were blocked into 6 groups. Cadmium chloride was given at levels of 0 or 325 mg (Cd: 0, 160 ppm), and chlorella powder at levels of 0, 3 and 5%. Cadmium was accumulated in blood and tissues (liver, kidney and small intestine) in the Cd exposed groups, while the accumulation of Cd was decreased in the Cd-exposed chlorella groups. Fecal and urinary Cd excretions were remarkably increased in Cd exposed chlorella groups. Thus, cadmium retention ratio and absorption rate were decreased in the Cd exposed chlorella groups. Urinary and serum creatinine, and creatinine clearance were not changed in experimental animals. In addition, metallothionein (MT) synthesis in tissues was increased by Cd administration. The Cd-exposed chlorella groups indicated lower MT concentration compared to the Cd exposed groups. Moreover, glomerular filtration rate (GFR) was not changed by dietary chlorella and Cd administration. According to the results above, this study could suggest that Cd toxicity can be alleviated by increasing Cd excretion through feces. Therefore, when exposed to Cd, chlorella is an appropriate source which counteracts heavy metal poisoning, to decrease the damage of tissues by decreasing cadmium absorption. PMID- 20016698 TI - Hypoglycemic effect of Chlorella vulgaris intake in type 2 diabetic Goto-Kakizaki and normal Wistar rats. AB - The aim of this study was to examine the hypoglycemic effect of chlorella in 6 week-old type 2 diabetic Goto-Kakizaki (GK, n=30) rats and 6 week-old normal Wistar (n=30) rats. Animals were randomly assigned to 3 groups respectively, and were fed three different experimental diets containing 0%, 3% or 5% (w/w) chlorella for 8 weeks. In diabetic GK rats, the insulinogenic-indices were not significantly different among the groups. The concentrations of fasting plasma glucagon and hepatic triglyceride, and the insulin/glucagon ratios of the GK-3% chlorella and GK-5% chlorella groups were significantly lower than those of the GK-control group. The HOMA-index and the concentrations of fasting blood glucose and plasma insulin of the GK-3% chlorella and GK-5% chlorella groups were slightly lower than those of the GK-control group. In normal Wistar rats, the insulinogenic-indices were not significantly different among the normal groups, but that of the Wistar-5% chlorella group was slightly higher than the other groups. The concentrations of fasting blood glucose and plasma insulin, and the HOMA-index of the Wistar-5% chlorella group were a little higher, and the fasting plasma glucagon concentration and the insulin/glucagon ratio of the Wistar-5% chlorella group were significantly higher than those of the Wistar-control and Wistar-3% chlorella groups. In conclusion, this study shows that the glucose stimulated insulin secretion was not affected by the intake of chlorella, which could be beneficial, however, in improving insulin sensitivity in type 2 diabetic GK and normal Wistar rats. PMID- 20016699 TI - The effect of seamustard on blood lipid profiles and glucose level of rats fed diet with different energy composition. AB - Recently, Korean people are consuming seaweeds almost 3.5 times more now than three decades ago. It is well known that seaweeds contain lots of soluble dietary fiber in addition to micronutrients such as beta-carotene, iodine and some bioactive components. Seaweeds are considered to be effective for preventing chronic diseases including obesity, diabetes mellitus, atherosclerosis, cancer or constipation. This study was conducted to investigate the effect of seamustard intake on body weight gain, blood glucose level and lipid profiles in rats fed diets with different energy nutrient composition. Male Sprague-Dawley rats (average initial weight 103.7 g) were divided into groups for two experiments as follows; Control, M2.5 & M5 groups (Exp. I) and M5, M10, HCM5, HCM10, HFM5 & HFM10 groups (Exp. II). The rats were fed diet and water ad libitum for 4 weeks. In general, there was no significant difference in blood glucose and triglyceride concentration among groups. In Exp. I, serum LDL-cholesterol level of rats fed diet with 5% seamustard powder (M5) was significantly lower than that of control group, while HDL-cholesterol level, TC/LDL ratio and weight of adrenal gland were higher. In Exp. II, food intake, body weight gain and EER of high fat diet with 10% seamustard group (HFM10) were the lowest among groups. Except gastrocnemius muscle, all organ weights of HFM10 group were the lowest. Fecal cholesterol excretion and serum LDL-cholesterol concentration of HFM10 group were the highest, while serum HDL-cholesterol level was the lowest among groups. Interestingly, HDL-cholesterol concentration was the highest in HCM5 group among groups. From these results, it was suggested that seamustard intake might be more effective for body weight control, but not for improving blood lipid profiles in high fat diet than in high carbohydrate diet. PMID- 20016700 TI - Effects of gender and gonadectomy on growth and plasma cholesterol levels in pigs. AB - We conducted two studies to determine the effect of gender, gonadectomy (GDX) on growth and plasma cholesterol levels in pigs. In experiment 1, five sham-operated and five GDX female Landrace pigs (26 kg) were allowed to have free access to water and feed up to market weight (approximately 100 kg). Body weight and feed consumption were recorded biweekly, and daily body weight gain, daily feed intake and feed efficiency (gain/feed) were calculated during the feeding period. In experiment 2, 10 male (26 kg) and 10 female (26 kg) Landrace pigs were used; five male and five female pigs were assigned to sham-operated or GDX. Pigs were allowed to have free access to water and a diet without added cholesterol (Table 1) until they were 6 months old (male 104 and female 98 kg) and thereafter they were fed a hypercholesterolemic diet (Table 1) containing 0.5% cholesterol and 0.1% cholate for 10 days. GDX of female pigs increased average daily gain (P<0.05), compared with their sham-operated counterparts during the growing finishing period, but had no effect (P>0.05) on feed efficiency. Plasma cholesterol levels in pigs fed a hypercholesterolemic diet for 10 days were much higher (P<0.05) in females than in males (161 vs 104 mg/100 mL plasma), and were increased by GDX only in male pigs. HDL-cholesterol/LDL+VLDL-cholesterol ratio appeared to be higher in males than in females, and was not influenced by GDX in either sex. Results suggested that the lower growth rate of female pigs than their male counterparts is attributable to the ovarian activity, and the lower plasma cholesterol level in male than in female pigs fed a hypercholesterolemic diet is due to the testicular activity. PMID- 20016701 TI - Folate nutrition is related to neuropsychological functions in the elderly. AB - We investigated the nutritional state of B vitamins and the neuropsychological functions in 25 subjects, aged 63.1 +/- 6.3 years, residing in rural areas of Korea. Nutritional states of thiamin, riboflavin, and pyridoxine were assessed enzymatically in the erythrocytes, and folate concentrations were measured microbiologically in the plasma and erythrocytes. A battery of composite neuropsychological test was administered to the subjects. Plasma folate was correlated with the total intelligence score (p=0.049). Folate levels in the erythrocytes were correlated with the performance intelligence scores such as block design (p=0.017) and picture arrangement (p=0.016). The red cell folate was correlated with memory scores such as general memory (p=0.009) and delayed recall (p=0.000). Although it did not reach statistical significance, verbal memory (p=0.053) was highly correlated with the red cell folate. The red cell folate was also correlated positively with the percent of conceptual level response number score (p=0.029), and negatively with the grooved pegboard test score for the non dominant hand (p=0.010). Fine motor coordination was also influenced by folate nutrition, as finger tapping scores in both hands were significantly correlated with red cell folate (dominant hand; p=0.026, non-dominant hand; p=0.004). Other B vitamins such as thiamin, riboflavin, and vitamin B(6) were not as strongly correlated with neuropsychological function test scores as folate was. These results suggest that folate nutrition influences neuropsychological function test scores significantly in humans. Further studies are needed to explore the relationship between folate or other vitamin B nutrition and neuropsychological functions and the implications thereof. PMID- 20016702 TI - Comparison of foodservice management performance level between dietitians and non dietitians in senior centers using IPA. AB - This study investigated the management importance and performance level of foodservice managers at senior centers. Using the survey, perceived importance and performance levels of seven foodservice management areas were evaluated and analyzed. Data showed the foodservice facilities were being managed by dietitians (61.6%) or non-dietitians (38.9%). The result indicated that overall importance level (3.43) was higher than performance level (3.02) (p<.01). As of the IPA result, dietitians and non-dietitians had different perspectives in terms of managing the eight categories of foodservice areas. The differences in the IPA results between the two groups may reflect bias attributable to the respondents' degrees of knowledge and professional preparation. The research findings could enhance our understanding of importance of hiring professional dietitians to operate foodservice at senior centers and find out which management area should be concentrated for more effective foodservice management. PMID- 20016703 TI - Benefits, barriers, self-efficacy and knowledge regarding healthy foods; perception of African Americans living in eastern North Carolina. AB - African Americans in the United States suffer from many health disparities such as obesity, diabetes or hypertension. Lifestyle factors including diet and physical activity play an important role in prevention of these health conditions. The purpose of this research project was to assess beliefs, barriers and self-efficacy of eating a healthy diet and self efficacy of shopping for foods such as whole grains or foods designated as low fat or low sodium. Additionally, the objective was to assess beliefs about healthfulness, appropriate consumption, and protective aspect of specific foods including fruits, vegetables, and whole grains. The assessment was done using a survey instrument developed for this study. Data collection took place at two church locations. Data were obtained from 57 African Americans, mean age 50 years old (SD 12.70) completed the survey. The majority of respondents (58.1%) were females and most (75%) had at least some college education. Generally, benefits of eating healthy foods received considerably higher scores compared to barriers of eating healthy. A belief that healthy foods would help to take care of one's body received the highest mean score while a belief that healthy foods are too expensive had the highest score from all barriers. The results showed high self efficacy of eating and purchasing healthy foods, high awareness of knowledge regarding foods associated with disease prevention but low awareness of recommendations for fruits and vegetables. The high scores for benefits, self efficacy and knowledge regarding eating healthy foods did not translate into the perception of intake of such foods. Most participants believed that they do not eat enough of healthy foods. Interventions design to help African Americans make dietary changes should be culturally relevant and should involved working on a community level utilizing messages that are familiar and relevant to African Americans. PMID- 20016704 TI - Mechanisms of amino acid sensing in mTOR signaling pathway. AB - Amino acids are fundamental nutrients for protein synthesis and cell growth (increase in cell size). Recently, many compelling evidences have shown that the level of amino acids is sensed by extra- or intra-cellular amino acids sensor(s) and regulates protein synthesis/degradation. Mammalian target of rapamycin complex 1 (mTORC1) is placed in a central position in cell growth regulation and dysregulation of mTOR signaling pathway has been implicated in many serious human diseases including cancer, diabetes, and tissue hypertrophy. Although amino acids are the most potent activator of mTORC1, how amino acids activate mTOR signaling pathway is still largely unknown. This is partly because of the diversity of amino acids themselves including structure and metabolism. In this review, current proposed amino acid sensing mechanisms to regulate mTORC1 and the evidences pro/against the proposed models are discussed. PMID- 20016705 TI - Effects of retinoic acid isomers on apoptosis and enzymatic antioxidant system in human breast cancer cells. AB - Retinoic acids (RAs) modulate growth, differentiation, and apoptosis in normal, pre-malignant & malignant cells. In the present study, the effects of RA isomers (all-trans RA, 13-cis RA, and 9-cis RA) on the cell signal transduction of human breast cancer cells have been studied. The relationship between RAs and an enzymatic antioxidant system was also determined. Estrogen-receptor (ER) positive MCF-7 and ER-negative MDA-MB-231 human breast cancer cells were treated with different doses of each RA isomers, all-trans RA, 13-cis RA, or 9-cis RA. Treatment of RA isomers inhibited cell viability and induced apoptosis of MCF-7 cells as a result of increased caspase activity in cytoplasm and cytochrome C released from mitochondria. All-trans RA was the most effective RA isomer in both cell growth inhibition and induction of apoptosis in MCF-7 cells. However, no significant effect of RA isomers was observed on the cell growth or apoptosis in ER-negative MDA-MB-231 cells. In addition, activities of antioxidant enzymes such as catalase and glutathione peroxidase were decreased effectively after treatment of RA in MCF-7 cells, whereas SOD activity was rarely affected. Thus, the present data suggest that all-trans RA is the most potential inducer of apoptosis and modulator of antioxidant enzymes among RA isomers in MCF-7 human breast cancer cells. PMID- 20016706 TI - The inhibition of inflammatory molecule expression on 3T3-L1 adipocytes by berberine is not mediated by leptin signaling. AB - In our previous study, we have shown that berberine has both anti-adipogenic and anti-inflammatory effects on 3T3-L1 adipocytes, and the anti-adipogenic effect is due to the down-regulation of adipogenic enzymes and transcription factors. Here we focused more on anti-inflammatory effect of berberine using real time RT-PCR and found it changes expressions of adipokines. We hypothesized that anti adipogenicity of berberine mediates anti-inflammtory effect and explored leptin as a candidate mediator of this signaling. We studied this hypothesis by western blot analysis, but our results showed that berberine has no effect on the phosphorylations of STAT-3 and ERK which have important roles on leptin signaling. These results led us to conclude that the anti-inflammatory effect of berberine is not mediated by the inhibition of leptin signal transduction. Moreover, we have found that berberine down-regulates NF-kappaB signaling, one of the inflammation-related signaling pathway, through western blot analysis. Taken together, the anti-inflammatory effect of berberine is not mediated by leptin, and berberine induces anti-inflammatory effect independent of leptin signaling. PMID- 20016707 TI - Effect of Chlorella vulgaris intake on cadmium detoxification in rats fed cadmium. AB - The aim of this study was to investigate if dietary Chlorella vulgaris (chlorella) intake would be effective on cadmium (Cd) detoxification in rats fed dietary Cd. Fourteen-week old male Sprague-Dawley (SD) rats weighing 415.0 +/- 1.6 g were randomly divided into two groups and fed slightly modified American Institute of Nutrition-93 Growing (AIN-93G) diet without (n=10) or with (n=40) dietary Cd (200 ppm) for 8 weeks. To confirm alteration by dietary Cd intake, twenty rats fed AIN-93G diet without (n=10) and with (n=10) dietary Cd were sacrificed and compared. Other thirty rats were randomly blocked into three groups and fed slightly modified AIN-93G diets replacing 0 (n=10), 5 (n=10) or 10% (n=10) chlorella of total kg diet for 4 weeks. Daily food intake, body weight change, body weight gain/calorie intake, organ weight (liver, spleen, and kidney), perirenal fat pad and epididymal fat pad weights were measured. To examine Cd detoxification, urinary Cd excretion and metallothonein (MT) concentrations in kidney and intestine were measured. Food intake, calorie intake, body weight change, body weight gain/calorie intake, organ weight and fat pad weights were decreased by dietary Cd intake. Urinary Cd excretion and MT concentrations in kidney and small intestine were increased by dietary Cd. After given Cd containing diet, food intake, calorie intake, body weight change, body weight gain/calorie intake, organ weights and fat pad weights were not influenced by dietary chlorella intake. Renal MT synthesis tended to be higher in a dose dependent manner, but not significantly. And chlorella intake did not significantly facilitate renal and intestinal MT synthesis and urinary Cd excretion. These findings suggest that, after stopping cadmium supply, chlorella supplementation, regardless of its percentage, might not improve cadmium detoxification from the body in growing rats. PMID- 20016708 TI - Diets with corn oil and/or low protein increase acute acetaminophen hepatotoxicity compared to diets with beef tallow in a rat model. AB - It has been reported that dietary polyunsaturated fats (PUFA) increase liver injury in response to ethanol feeding. We tested the hypothesis that diets rich in linoleic acid (18:2n-6) would affect acute liver injury after acetaminophen injection and that protein restriction might exacerbate the liver injury. We examined effects of feeding diets with either 15% (wt/wt) corn oil or 14% beef tallow and 1% corn oil for six weeks with either 6 or 20 g/100 g protein on acute hepatotoxicity. After the feeding period, liver injury was induced by injecting either with 600 mg/kg body weight acetaminophen suspended in gum arabic-based vehicle, or with vehicle alone during fasting status. Samples of liver and plasma were taken for analyses of hepatic glutathione (GSH) levels and liver-specific enzymes [(Glutamate-pyruvate transaminase (GPT) and glutamate-oxaloacetate transaminase (GOT)], respectively. Whereas GSH level was significantly lower in only group fed 15% corn oil with 6 g/100 g protein among acetaminophen-treated groups, activities of GPT and GOT were significantly elevated in all groups except the one fed beef tallow with 20 g/100 g protein, suggesting low protein might exacerbate drug-induced hepatotoxicity. The feeding regimens changed the ratio of 18:2n-6 to oleic acid (18:1n-9) in total liver lipids approximately five fold, and produced modest changes in arachidonic acid (20:4n-6). We conclude that diets with high 18:2n-6 promote acetaminophen-induced liver injury compared to diets with more saturated fatty acids (SFA). In addition, protein restriction appeared to exacerbate the liver injury. PMID- 20016709 TI - Effects of dietary fish oil and trans fat on rat aorta histopathology and cardiovascular risk markers. AB - Fish oil and shortening have been suggested to have opposite effects on cardiovascular disease (CVD). This study investigated the effect of shortening and fish oil on CVD risk factors and aorta histopathology, and the association between risk factors and aorta histopathology. Male Wister rats (n=30) were fed an AIN-93G diet containing 20% fat in the form of fish oil, shortening, or soybean oil for 4 weeks. Total cholesterol (TC), triacylglyceride (TG), and C reactive protein levels were significantly (P<0.001) lower in the fish oil than in soybean oil and shortening groups. HDL-cholesterol concentrations were significantly different (P<0.001) between groups. In addition, LDL-cholesterol levels were significantly (P<0.001) lower in the fish oil and shortening groups than in the soybean oil group. Insulin and glucose concentrations did not differ among groups. Effect of dietary fat on tissue fatty acid composition significantly differed in abdominal fat and brain compared with RBC, heart, kidney and liver. The aortic wall was significantly (P=0.02) thinner in the fish oil group than in the soybean oil and shortening groups. The aortic wall thickness was positively correlated with TG and TC, but negatively with EPA + DHA levels of all tissues. These results suggested that fish oil had protective effects on aorta histopathology by hypolipidemic action in this rat model. PMID- 20016710 TI - Effects of taurine supplementation on bone mineral density in ovariectomized rats fed calcium deficient diet. AB - Taurine supplementation has been shown to have a beneficial effect on femur bone mineral content in ovariectomized rats. It therefore seemed desirable to find out whether the beneficial effect of taurine on ovariectomized rats fed calcium deficient diet could also be reproduced. Forty female Sprague-Dawley rats were divided into two groups. One group was OVX and the other group received sham operation (SHAM), and received either control diet or a taurine supplemented diet for 6 weeks. All rats were fed on calcium deficient diet (AIN-93: 50% level of calcium) and deionized water. Bone mineral density (BMD) and bone mineral content (BMC) were measured in spine and femur. The serum and urine concentrations of calcium and phosphorus were determined. Bone formation was measured by serum osteocalcin and alkaline phosphatase (ALP) concentrations. Bone resorption rate was measured by deoxypyridinoline (DPD) crosslinks immunoassay and corrected for creatinine. Urinary calcium and phosphorus excretion, osteocalcin in blood and cross link value were not significantly different among the groups. Within the OVX group, the taurine supplemented group had not higher femur bone mineral content than the control group. This study established the need for a study on the taurine effect on bone with different calcium levels. PMID- 20016711 TI - Red beet (Beta vulgaris L.) leaf supplementation improves antioxidant status in C57BL/6J mice fed high fat high cholesterol diet. AB - The effect of diet supplemented with red beet (Beta vulgaris L.) leaf on antioxidant status of plasma and tissue was investigated in C57BL/6J mice. The mice were randomly divided into two groups after one-week acclimation, and fed a high fat (20%) and high cholesterol (1%) diet without (control group) or with 8% freeze-dried red beet leaf (RBL group) for 4 weeks. In RBL mice, lipid peroxidation determined as 2-thiobarbituric acid-reactive substances (TBARS value) was significantly reduced in the plasma and selected organs (liver, heart, and kidney). Levels of antioxidants (glutathione and beta-carotene) and the activities of antioxidant enzyme (glutathione peroxidase) in plasma and liver were considerably increased, suggesting that antioxidant defenses were improved by RBL diet. Comet parameters such as tail DNA (%), tail extent moment, olive tail moment and tail length were significantly reduced by 25.1%, 49.4%, 35.4%, and 23.7%, respectively, in plasma lymphocyte DNA of RBL mice compared with control mice, and indicated the increased resistance of lymphocyte DNA to oxidative damage. In addition, the RBL diet controlled body weight together with a significant reduction of fat pad (retroperitoneal, epididymal, inguinal fat, and total fat). Therefore, the present study suggested that the supplementation of 8% red beet leaf in high fat high cholesterol diet could prevent lipid peroxidation and improve antioxidant defense system in the plasma and tissue of C57BL/6J mice. PMID- 20016712 TI - Effects of dietary supplementation of high-dose folic acid on biomarkers of methylating reaction in vitamin B(12)-deficient rats. AB - Folate is generally considered as a safe water-soluble vitamin for supplementation. However, we do not have enough information to confirm the potential effects and safety of folate supplementation and the interaction with vitamin B(12) deficiency. It has been hypothesized that a greater methyl group supply could lead to compensation for vitamin B(12) deficiency. On this basis, the present study was conducted to examine the effects of high-dose folic acid (FA) supplementation on biomarkers involved in the methionine cycle in vitamin B(12)-deficient rats. Sprague-Dawley rats were fed diets containing either 0 or 100 microg (daily dietary requirement) vitamin B(12)/kg diet with either 2 mg (daily dietary requirement) or 100 mg FA/kg diet for six weeks. Vitamin B(12) deficiency resulted in increased plasma homocysteine (p<0.01), which was normalized by dietary supplementation of high-dose FA (p<0.01). However, FA supplementation and vitamin B(12) deficiency did not alter hepatic and brain S adenosylmethionine (SAM) and S-adenosylhomocysteine (SAH) concentrations and hepatic DNA methylation. These results indicated that supplementation of high dose FA improved homocysteinemia in vitamin B(12)-deficiency but did not change SAM and SAH, the main biomarkers of methylating reaction. PMID- 20016713 TI - Comparison of time course changes in blood glucose, insulin and lipids between high carbohydrate and high fat meals in healthy young women. AB - Few studies have examined short term responses to the different contents of carbohydrate or fat in the meal, although long term effects of the high fat meal have been considered as compound risk factor for metabolic disorders. The aim of this study was to investigate the postprandial changes of plasma glucose, insulin and lipids upon intakes of high carbohydrate or high fat meal in young healthy women. Subjects were randomly assigned to either the high carbohydrate meal (HCM, 75% carbohydrate, n=13) or the high fat meal (HFM, 60% fat, n=12) groups. The meals were prepared as isocaloric typical Korean menu. Blood samples were obtained prior to and 30, 60, 90, 120, 180 and 240 minute after the meal. There were no significant differences on fasting blood parameters including glucose, insulin, lipids concentrations between the groups prior to the test. The HCM had higher blood glucose and insulin concentrations, reached the peak at 30 min and maintained for 240 min compared to the HFM (P<0.05). The HFM had higher plasma triglyceride (TG) and free fatty acid (FFA) concentrations, reached the peak at 120 min and maintained for 240 min compared to the HCM (P<0.05). It is concluded that macronutrients content in the meal may be an important determinant of postprandial substrate utilization in healthy women. PMID- 20016715 TI - Relationships of family support, diet therapy practice and blood glucose control in typeII diabetic patients. AB - The purpose of this study was to investigate the degree of family support for diabetic patients and the diet therapy practice of patients themselves, and to analyze the relationship between family support and diet therapy practice and blood glucose control, and thus to prepare basic data for the development of effective education programs to improve blood glucose control in diabetic patients. The study subjects were 82 patients with type II diabetes, aged over 20 in the Chungbuk area. The gender distribution of subjects was 52.4% males and 47.6% females, and BMI showed 29.3% overweight and 35.3% obesity. Among the 82 study subjects, the relationship between diet therapy related family support and blood glucose control was examined in 67 subjects who answered practicing diet therapy, and the results showed that the family support score of a group with excellent blood glucose control was significantly higher than those of groups with fair or poor control (p<0.001) and the correlation between the two factors was very high (r=0.341, p<0.001). For the relationship between diet therapy practice by patients themselves and blood glucose control, diet therapy practice of a group with excellent blood glucose control was significantly higher than those of other groups (fair or poor control groups) (p<0.001) and the correlation between two factors was very high (r=0.304, p<0.001). For other factors influencing blood glucose control, a group with diabetes education showed significantly better blood glucose control compared to other groups without education (p<0.05). From the above results, diet therapy practice by patients, family support, and the necessity of diabetes education were confirmed to control blood glucose of diabetic patients. In conclusion, development and operation of education program should include not only patients but also their family members. PMID- 20016714 TI - The factors affecting pregnancy outcomes in the second trimester pregnant women. AB - Adequate nutrient intake during pregnancy is important to fetal and maternal health. The purpose of this study was to investigate the factors affecting birth weight and gestational age and to provide basic data to promote more favorable pregnancy outcomes. Data were collected from 234 pregnant women at two hospitals in Seoul. Demographic characteristics, anthropometric measurements and health related habits were obtained using a questionnaire at the hospital visit during the second trimester. Dietary intakes were estimated by 24 hour recall at the hospital visit during the second trimester. Data on pregnancy outcomes, including birth weights and gestational ages, were obtained from hospital records after delivery. Birth weights were divided into a low birth weight group (birth weight<3.1 kg), a normal birth weight group (3.1-3.6 kg) and a high birth weight group (>3.6 kg). Gestational ages were divided into tertiles according to the gestational age of the subjects: group 1 (<38.53 weeks), group 2 (38.53-40.00 weeks) and group 3 (>40.00 weeks). The number of family members was significantly lower in the low birth weight group than in the normal birth weight group (p<0.05). In the low birth weight group, pregnancy weight was significantly lower than in the high birth weight group (p<0.05). Health related habits were not significantly different among any of the groups. Intakes of fiber, phosphorous, iron, vitamin B(6) and folic acid were significantly higher in the high birth weight group than the low birth weight group (p<0.05). Gestational age was not significantly affected by nutrient intakes, but birth weight was affected by nutrient intake in the results of this study. Therefore, the adequacy of nutrient intake is important for the improvement of pregnancy outcomes. PMID- 20016716 TI - Antioxidant effect of garlic and aged black garlic in animal model of type 2 diabetes mellitus. AB - Hyperglycemia in the diabetic state increases oxidative stress and antioxidant therapy can be strongly correlated with decreased risks for diabetic complications. The purpose of this study is to determine antioxidant effect of garlic and aged black garlic in animal model of type 2 diabetes. The antioxidant activity of garlic and aged black garlic was measured as the activity in scavenging free radicals by the trolox equivalent antioxidant capacity (TEAC) assay. Three week-old db/db mice were fed AIN-93G diet or diet containing 5% freeze-dried garlic or aged black garlic for 7 weeks after 1 week of adaptation. Hepatic levels of lipid peroxides and activities of antioxidant enzymes were measured. TEAC values of garlic and aged black garlic were 13.3 +/- 0.5 and 59.2 +/- 0.8 micromol/g wet weight, respectively. Consumption of aged black garlic significantly decreased hepatic thiobarbituric acid reactive substances (TBARS) level compared with the garlic group which showed lower TBARS level than control group (p<0.05). Activities of superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) of garlic and aged black garlic group were significantly elevated compared to the control group. Catalase (CAT) activity of aged black garlic group was increased compared with the control group. These results show that aged black garlic exerts stronger antioxidant activity than garlic in vitro and in vivo, suggesting garlic and aged black garlic, to a greater extent, could be useful in preventing diabetic complications. PMID- 20016717 TI - Dietary patterns associated with hypertension among Korean males. AB - The objectives of this study were to identify the dietary patterns associated with hypertension among Korean males. Data from the 2001 Korean National Health and Nutrition Survey of 1,869 men aged 20-65 years were used for the analysis. As an initial analysis, a factor analysis was applied to identify major dietary patterns among the subjects. Then logistic regression analysis was conducted to identify the pattern related with hypertension. As a result of the initial analysis, three major dietary patterns were identified. Dietary pattern 1 (traditional) was heavily loaded with vegetables, fish and cereal. Dietary pattern 2 (Western) was loaded with fast foods, bread, meats and dairy products. Dietary pattern 3 (Drinker) was loaded with mostly pork, beer and soju (Korean liquor). From the second stage of the analysis, there was a tendency of positive association between traditional patterns and hypertension risks. However, the tendency did not meet statistical significance level (p<0.05). In summary, unlikely findings from European and American studies, vegetables rich traditional dietary patterns did not show any protective effect on hypertension in Korean males. The Korean dietary practice, which is consuming salted vegetables instead of fresh vegetables, might have played a role in these findings. However, the full explanation of the findings remained to be answered with further investigation since none of the dietary patterns identified showed any statistical significance. PMID- 20016718 TI - In vitro inhibition of 10-formyltetrahydrofolate dehydrogenase activity by acetaldehyde. AB - Alcoholism has been associated with folate deficiency in humans and laboratory animals. Previous study showed that ethanol feeding reduces the dehydrogenase and hydrolase activity of 10-formyltetrahydrofolate dehydrogenase (FDH) in rat liver. Hepatic ethanol metabolism generates acetaldehyde and acetate. The mechanisms by which ethanol and its metabolites produce toxicity within the liver cells are unknown. We purified FDH from rat liver and investigated the effect of ethanol, acetaldehyde and acetate on the enzyme in vitro. Hepatic FDH activity was not reduced by ethanol or acetate directly. However, acetaldehyde was observed to reduce the dehydrogenase activity of FDH in a dose- and time-dependent manner with an apparent IC(50) of 4 mM, while the hydrolase activity of FDH was not affected by acetaldehyde in vitro. These results suggest that the inhibition of hepatic FDH dehydrogenase activity induced by acetadehyde may play a role in ethanol toxicity. PMID- 20016719 TI - Pancreatic lipase inhibitory activity of taraxacum officinale in vitro and in vivo. AB - Obesity has become a worldwide health problem. Orlistat, an inhibitor of pancreatic lipase, is currently approved as an anti-obesity drug. However, gastrointestinal side effects caused by Orlistat may limit its use. In this study the inhibitory activities of dandelion (Taraxacum officinale) against pancreatic lipase in vitro and in vivo were measured to determine its possible use as a natural anti-obesity agent. The inhibitory activities of the 95% ethanol extract of T. officinale and Orlistat were measured using 4-methylumbelliferyl oleate (4 MU oleate) as a substrate at concentrations of 250, 125, 100, 25, 12.5 and 4 microg/ml. To determine pancreatic lipase inhibitory activity in vivo, mice (n=16) were orally administered with corn oil emulsion (5 ml/kg) alone or with the 95% ethanol extract of T. officinale (400 mg/kg) following an overnight fast. Plasma triglyceride levels were measured at 0, 90, 180, and 240 min after treatment and incremental areas under the response curves (AUC) were calculated. The 95% ethanol extract of T. officinale and Orlistat, inhibited, porcine pancreatic lipase activity by 86.3% and 95.7% at a concentration of 250 microg/ml, respectively. T. officinale extract showed dose-dependent inhibition with the IC(50) of 78.2 microg/ml. A single oral dose of the extract significantly inhibited increases in plasma triglyceride levels at 90 and 180 min and reduced AUC of plasma triglyceride response curve (p<0.05). The results indicate that T. officinale exhibits inhibitory activities against pancreatic lipase in vitro and in vivo. Further studies to elucidate anti-obesity effects of chronic consumption of T. officinale and to identify the active components responsible for inhibitory activity against pancreatic lipase are necessary. PMID- 20016720 TI - Effect of Chlorella vulgaris on lipid metabolism in Wistar rats fed high fat diet. AB - This study was performed to investigate effects of Chlorella vulgaris on lipid metabolism in rats fed high fat diet. Sixty 6-week-old male Wistar rats were divided into two groups; normal diet group and high fat diet group, then the rats in each group were further divided into three subgroups and fed 0%, 5% and 10% (w/w) chlorella-containing diets, respectively, and raised for 9 weeks. Serum aspartate aminotransferase (AST) and alanine aminotransferase (ALT) activity and total protein and albumin concentration were not different among groups. Serum total lipids and liver TG concentration were significantly lower in 5% and 10% chlorella groups than 0% chlorella group in high fat diet groups (p<0.05). Serum TG, serum total cholesterol, liver total lipid and liver total cholesterol concentrations were significantly lower in 10% chlorella groups than 0% chlorella group in high fat diet groups (p<0.05). Fecal total lipid, TG and total cholesterol excretions were significantly higher in 5% and 10% chlorella groups than 0% chlorella groups in normal diet and high fat diet groups, respectively (p<0.05). These results suggest that Chlorella vulgaris is effective for prevention of dyslipidemia which may be due to the modulation of lipid metabolism and increased fecal excretion of lipid. PMID- 20016721 TI - Effect of onion and beet on plasma and liver lipids, platelet aggregation, and erythrocyte Na efflux in simvastatin treated hypercholesterolmic rats. AB - This study was purposed to investigate the effect of onion or beet on plasma and liver lipids, erythrocyte Na efflux channels and platelet aggregation in simvastatin (SIM) treated hypercholesterolemic rats. Forty Sprague Dawley rats were divided into four groups and fed 0.5% cholesterol based diets containing 2 mg/kg BW simvastatin or simvastatin with 5% onion or beet powder. Plasma total cholesterol was significantly increased in SIM group compared with the control (p<0.01), and the elevated plasma total cholesterol of SIM group was significantly decreased in SIM-onion and SIM-beet groups (p<0.05). HDL cholesterol in SIM-beet group was significantly increased compared with other groups (p<0.05). Platelet aggregation in both the maximum and initial slope was significantly decreased in SIM group compared with SIM-onion group (p<0.05). Na-K ATPase was significantly decreased in SIM group compared with the control, SIM onion and SIM-beet groups (p<0.05). Na passive leak was significantly increased in all groups treated with SIM compared with the control (p<0.05). The total Na efflux was decreased in SIM group and increased in SIM-onion group and the difference between these two groups was significant (p<0.05). There was no difference in intracellular Na among groups. In present study, simvastatin, a HMG CoA reductase inhibitor at dose of 2mg/kg BW/day rather increased plasma total cholesterol in rats, inferring that the action mechanism of simvastatin on cholesterol metabolism differ between rat and human. Onion and beet play favorable roles in cardiovascular system by restoring the reduced Na efflux through Na-K ATPase and Na-K cotransport in SIM treated rats. PMID- 20016722 TI - Effects of soybean isoflavone extract on the plasma lipid profiles and antioxidant enzyme activity in streptozotocin-induced diabetic rats. AB - The present study evaluated the effects of various dosages of soybean isoflavone extract on lipid profiles, lipid peroxidation and antioxidant activities in streptozotocin-induced diabetic rats. The one normal control group was fed an AIN 76-based experimental diet and four diabetic groups were fed the same diet, supplemented with four different levels of soybean isoflavone extract for seven weeks. The daily dosages of pure isoflavone for four diabetic groups were set to be 0 mg (diabetic control), 0.5 mg (ISO-I), 3.0 mg (ISO-II) and 30.0 mg (ISO-III) per kilogram of body weight, respectively. The plasma total cholesterol levels and the TBA-reactive substances contents in the liver and kidney were significantly lowered in ISO-II and ISO-III groups compared to those in the diabetic control group. The levels of plasma HDL-cholesterol, plasma vitamin A and hepatic superoxide dismutase were significantly increased in those two groups compared with the diabetic control group. The present study demonstrated the possibility that the diets supplemented with 3.0 mg and 30.0 mg of soybean isoflavone extract may have beneficial effects on the plasma lipids, tissue lipid peroxidation and partly on antioxidant system in diabetic animals and there were no significant differences between the ISO-II and ISO-III groups. The results suggest that the effective daily dosage level of isoflavone for improving lipid metabolism in diabetic rats may be above 3.0 mg per kilogram body weight. PMID- 20016723 TI - Grape seed extract (Vitis vinifera) partially reverses high fat diet-induced obesity in C57BL/6J mice. AB - The aim of the present study was to assess the anti-obesity effects of grape seed extract (GSE) supplement in C57BL/6J mice. Thirty mice were divided into three groups; normal diet control group (ND), high fat diet control group (HD) and high fat diet plus grape seed extract supplemented group (HD+GSE). Results were as follows: 1. GSE supplement reduced the weight gain in mice fed high fat diets; epididymal and back fat weights were lower compared to non-supplemented HD group. 2. Blood lipid concentrations were lower in the HD+GSE group than in the HD group. Serum HDL-C concentrations were higher in the HD+GSE group compared with the other groups. 3. The concentrations of acid-insoluble acylcarnitines (AIAC) in serum and liver were higher in the HD+GSE group than in the HD group. 4. GSE supplementation increased mRNA levels of lipolytic genes such as carnitine palmitoyltransferase-1 (CPT-1) and decreased mRNA levels of lipogenic genes such as acetyl CoA carboxylase (ACC). These findings suggest that grape seed extract supplements in high fat diet might normalize body weight, epididymal and back fat weights, lipid concentrations, and carnitine levels through controlling lipid metabolism. PMID- 20016724 TI - Pre- and post-initiation modulating effects of green tea ingestion on rat hepatocarcinogenesis. AB - The purpose of this study was to investigate the effects of green tea ingestion on hepatocarcinogenesis before and after its initiation. Male Sprague-Dawley rats were fed an AIN76A diet with or without green tea. Initiation was induced by a single dose (200 mg/kg) of diethylnitrosamine at week 4 and 0.02% (w/w) 2 acetylaminofluorene was supplied in the diets. The control group had free access to water for 13 weeks (CTR13). Tea infusion was provided from the beginning of the experiment for 13 weeks (PRE13) or from the post-initiation stage until week 13 (POST13). Three other groups (CTR24, PRE24 and POST24) were added to examine the longer-term effects (24 weeks) with the same experimental design. The percentage area of liver sections that were positive for hepatic placental glutathione S-transferase (GST-P), which was used as a marker of preneoplastic lesions, was smaller in PRE13 (20.2 +/- 5.0%, mean +/- SD) and POST13 (26.0 +/- 4.8%) than in CTR13 (33.2 +/- 5.8%, p<0.05). Over the longer period, the GST-P lesions were significantly smaller for both PRE24 and POST24 (21.6 +/- 8.5% and 22.2 +/- 4.0%, respectively) than for CTR24 (28.6 +/- 5.1%, p<0.05), but there was no significant difference between PRE24 and POST24. The liver content of thiobarbituric acid reactive substances was significantly lower in the tea groups than in the controls (p<0.05). However, no significant differences were observed among groups of GST activity. The results show that tea consumption exhibits a stronger short-term initiation-inhibiting ability in liver carcinogenesis, but over a longer period, the preventive effects of green tea ingestion do not differ in post- and pre-initiation. PMID- 20016725 TI - Effect of butanol fraction from Cassia tora L. seeds on glycemic control and insulin secretion in diabetic rats. AB - Cassia tora L. seeds have previously been reported to reduce blood glucose level in human and animals with diabetes. In the present study, the effects of Cassia tora L. seed butanol fraction (CATO) were studied on postprandial glucose control and insulin secretion from the pancreas of the normal and diabetic rats. Diabetes was induced by an i.p. injection of Streptozotocin (55 mg/kg BW) into the male Sprague-Dawley rats. The postprandial glucose control was monitored during a 240 min-period using a maltose loading test. In normal rats, rats fed CATO (20 mg/100 g BW/d) showed lower postprandial glucose levels in all the levels from 30 min up to 180 min than those in the control rats without CATO (p<0.05). In diabetic rats, those levels in the CATO group seemed to be lower during the 30~180 min, but only glucose level at 30 min showed significant difference compared to that in the control group. Moreover, CATO delayed the peak time of the glucose rise in both normal and diabetic rats in the glucose curves. On the other hand, when CATO was administered orally to the diabetic rats for 5 days, 12 hr fasting serum glucose level was decreased in the diabetic rats (p<0.05). Degree of a decrease in 12 hr fasting serum insulin levels was significantly less in the diabetic CATO rats as compared to diabetic control rats. On the last day of feeding, beta cells of the pancreas were stimulated by 200 mg/dL glucose through a 40 min-pancreas perfusion. Amounts of the insulin secreted from the pancreas during the first phase (11~20 min) and the second phase (21~40 min) in the CATO fed diabetic rats were significantly greater than those in the diabetic control group (p<0.05). These findings indicated that constituents of Cassia tora L. seeds have beneficial effect on postprandial blood glucose control which may be partially mediated by stimulated insulin secretion from the pancreas of the diabetic rats. PMID- 20016726 TI - Antioxidant and antidiabetic activities of extracts from Cirsium japonicum roots. AB - This study investigated the antioxidant activity of methanol (MeOH) and water extracts from roots of Cirsium japonicumin vitro. MeOH extract showed a stronger free radical scavenging activity than water extract. However, both of extracts showed a concentration dependent hydroxyl radical scavenging activity, reducing power and metal chelating ability. MeOH extract had greater phenolic and flavonoid contents than water extract. The antidiabetic activity of these two extracts was evaluated by the alpha-glucosidase inhibition assay. The water extract showed a considerable alpha-glucosidase inhibitory activity. To our knowledge, this may be the first time to report the antioxidant and antidiabetic activities in Cirsium japonicum roots. PMID- 20016727 TI - Effect of fructose or sucrose feeding with different levels on oral glucose tolerance test in normal and type 2 diabetic rats. AB - This study was designed to determine whether acute fructose or sucrose administration at different levels (0.05 g/kg, 0.1 g/kg or 0.4 g/kg body weight) might affect oral glucose tolerance test (OGTT) in normal and type 2 diabetic rats. In OGTT, there were no significant differences in glucose responses between acute fructose- and sucrose-administered groups. However, in normal rats, the AUCs of the blood glucose response for the fructose-administered groups tended to be lower than those of the control and sucrose-administered groups. The AUCs of the lower levels fructoseor sucrose-administered groups tended to be smaller than those of higher levels fructose- or sucrose-administered groups. In type 2 diabetic rats, only the AUC of the lowest level of fructose-administered (0.05 g/kg body weight) group was slightly smaller than that of the control group. The AUCs of fructose-administered groups tended to be smaller than those of the sucrose-administered groups, and the AUCs of lower levels fructose-administered groups tended to be smaller than those fed higher levels of fructose. We concluded from this experiment that fructose has tendency to be more effective in blood glucose regulation than sucrose, and moreover, that smaller amount of fructose is preferred to larger amount. Specifically, our experiments indicated that the fructose level of 0.05 g/kg body weight as dietary supplement was the most effective amount for blood glucose regulation from the pool of 0.05 g/kg, 0.1 g/kg and 0.4 g/kg body weights. Therefore, our results suggest the use of fructose as the substitute sweetener for sucrose, which may be beneficial for blood glucose regulation. PMID- 20016728 TI - Mothers' perceptions of children's food behaviors: use of focus group interview study. AB - Children's food behaviors have been expressed in a various ways because of recent changes in their family environment. Thus, this study was performed to investigate in-depth qualitative research on the mother's perception on children's food behaviors by focused group interview. This study was designed in four steps of planning, collection of participants, process, and analysis. Participants for the focus group interview were recruited and sampled from households with elementary school students in the Seoul and Gyeonggido areas. Groups were divided by total income and education expense levels. 1) High income household: It is better to improve currently existing web sites for nutrition education. 2) Mid income household: Easy, practical, and inexpensive off-line cooking class/nutrition education classes for mothers are needed. Nutrition programs for children should be developed through mass media and be promoted in the broadcasting circle. 3) Low income household: Motivation is required for mothers' education and the serious nutritional problems of children should be informed through mass media and home correspondence from school. And interesting educational materials should be developed for children to read whenever they want. PMID- 20016729 TI - Acculturation, meal frequency, eating-out, and body weight in Korean Americans. AB - Consuming regular meals has been studied in relation to better health, while higher regularity of eating-out has been linked to obesity. This study examined whether acculturation was associated with regularity of meals, eating-out, and overweight in Korean Americans. Pre-tested questionnaires were mailed to a U.S. national sample with Korean American surnames, and 55% of the deliverable sample responded, producing 356 usable questionnaires. Acculturation was measured using a two-culture matrix model and Gordon's theoretical work, and showed there were three distinct groups (acculturated, bicultural, and traditional). Only 36% reported that they regularly ate three meals a day. Breakfast was the least frequent meal of the day with 43% reporting eating breakfast everyday. More than half (58%) reported that they usually eat out or get take-out food at least once a week. After controlling for age, sex, income, education, and working status, higher acculturation was related to greater regularity of eating-out, but not meal regularity. A total of 28% of men and 6% of women were overweight (BMI>25), and there were significant and positive relationships between body weight status and acculturation in men but not women. However, no significant relationships between frequency of meals and eating-out and overweight status were present. This study did not find significant relationships of meal regularity and eating out with body weight, however, given the positive relationship between acculturation and eating-out among the subjects and the well-established relationship between eating-out and obesity, nutrition education about skipping meals and eating-for Korean Americans may be useful to prevent such relationships from developing. PMID- 20016730 TI - Soy food intake behavior by socio-demographic characteristics of Korean housewives. AB - In this study, the soy food intake behaviors including perception and intake frequency of soybean foods by sociodemographic variables were analyzed in housewives. The perception of housewives for soy foods showed that soybean paste, soybean curd, and Dambuk were high in the descending order for nutritional quality and health promotion effect, and soybean paste received the highest score in taste and flavor. Soybean sprouts received the highest evaluation score in the economic aspect. In the aspect of safe food, soybean paste received the highest evaluation score, as mush as a traditional food. The analysis of perception by sociodemographic variables showed that soybean curd, Dambuk, and soybean sprouts had higher perceptions as education level increased, and soy milk had higher perceptions in subjects with younger age and with employment. In the intake frequency, more than 50% of the subjects had soybean curd, soybean sprouts, and soybean paste more than once a week. The analysis for correlation between the intake frequency of soy foods and the degree of perception showed that taste and flavor had high correlation with the intake frequency of soy foods except soybean sprouts. The intake frequency of soybean paste, Dambuk, and soy milk had positive correlations to familiarity and that of soy milk had positive correlations to nutrition and health perception, and those of soybeam paste, soybean sprouts, and soy milk had positive correlations to safe food perception. From the above results, housewives in Korea had very high perceptions to nutritional quality and health promotion effect of soy foods and the degree of perception and accompanied intake frequency had significant differences by age, education level, and economic level among sociodemographic variables. PMID- 20016731 TI - Glycemic control of type 2 diabetic patients after short-term zinc supplementation. AB - This study was carried out to determine whether a short-term zinc supplementation contributes to beneficial changes in glycemic control among type 2 diabetic patients. Seventy-six diabetic subjects and 72 normal adults participated in this study. Subjects were divided into supplemented and control groups. Forty-four diabetic patients and 34 normal subjects were supplemented with 50 mg zinc daily as zinc gluconate for 4 weeks. Zinc status was assessed from fasting plasma levels and urinary excretion. The effects of zinc supplementation on fasting blood glucose, HbA(1c), insulin, and C-peptide were measured at the beginning of the study and after 4 weeks of supplementation. The changes in glycemic control indicators were compared between diabetic groups, classified by baseline HbA(1c) levels, and by diabetic duration. At baseline, the incidence of marginal zinc deficiency in the diabetic group, as determined by plasma zinc level, was approximately twice as high as in the normal adult group. The changes of HbA(1c) concentration, and fasting blood glucose following supplementation were not statistically significant in diabetic subjects. In normal subjects, a significant decrease of HbA(1c) occurred only in the zinc supplemented group. No significant changes were observed for serum insulin and C-peptide in diabetic as well as normal subjects. However, when the changes were compared by baseline HbA(1c) level, we found that diabetic subjects with HbA(1c) >/= 7.5% showed significantly improved levels of HbA(1c) and fasting glucose after Zn supplementation. While such improvement in fasting blood glucose was significant among diabetics with shorter diabetic duration, significant levels of increase in serum insulin and C peptide were observed in zinc supplemented subjects with longer diabetic duration. Fasting blood glucose was significantly decreased, whereas serum insulin and C-peptide were increased in diabetics with marginal zinc status. Therefore, we suggest that Zn supplementation for a short-term period may improve glycemic control in diabetic patients with higher HbA(1c) levels and marginal zinc status. PMID- 20016732 TI - The association of PBX1 polymorphisms with overweight/obesity and metabolic alterations in the Korean population. AB - Pre-B-cell leukemia transcription factor 1 (PBX1), which is located on chromosome 1q23, was recently reported to be associated with type 2 diabetes mellitus. We examined whether single nucleotide polymorphisms (SNPs) of the PBX1 gene are associated with overweight/obesity in a Korean population. We genotyped 66 SNPs in the PBX1 gene and investigated their association with clinical phenotypes found in 214 overweight/obese subjects and 160 control subjects using the Affymetrix Targeted Genotyping chip array. Seven SNPs (g.+75186C>T, g.+78350C>A, g.+80646C>T, g.+138004C>T, g.+185219G>A, g.+191272A>C, and g.+265317T>A) were associated with the risk of obesity in three models (codominant, dominant, and recessive) (P=0.007-0.05). Haplotype 1 (CAC) and 3 (TAC) of block 3 and haplotype 2 (GGAAT) of block 10 were also strongly associated with the risk of obesity. In the control group, subjects that had homozygote for the major allele for both g.+185219G>A and g.+191272A>C showed lower high density lipoprotein-cholesterol (HDL-C) level compared to those possessing the minor allele, suggesting that the association between the homozygote for the major allele for both g.+185219G>A and g.+191272A>C and HDL-C is attributable to the increased risk of obesity. This study suggests that the PBX1 gene is a possible risk factor in overweight/obese patients. PMID- 20016733 TI - A randomized study to establish the effects of spirulina in type 2 diabetes mellitus patients. AB - Spirulina is a microscopic and filamentous cyanobacterium that contains essential amino acids, essential fatty acids, vitamins, minerals and anti-oxidative components. The purpose of this study was to examine effects of spirulina intervention in Korean patients with type 2 diabetes. The subjects were 37 type 2 diabetic patients who visited a diabetic clinic in Seoul and randomly assigned into spirulina (8 g/day) or control group. During the intervention period of 12 weeks, subjects were asked to keep usual diet and prohibited to take any functional foods or dietary supplements. Spirulina supplementation for 12 weeks did not affect anthropometric parameters, however, lowered plasma triglycerides level significantly (p<0.05). Spirulina supplementation also resulted in a significant reduction in plasma malondialdehyde level (p<0.05) and an increase in plasma adiponectin level (p<0.1). The lipid lowering effect of spirulina supplementation was different according to serum lipid levels of the subjects before entering the intervention. The subjects with higher initial triglyceride level showed higher reduction in plasma triglyceride and blood pressure. The subjects with higher initial total cholesterol and LDL-cholesterol level showed higher reduction in plasma concentrations of total cholesterol, LDL-cholesterol, IL-6, and blood pressure. It seems that spirulina supplementation is more effective in subjects with dyslipidemia. This study provides the evidence for beneficial effects of spirulina supplementation on blood lipid profiles, inflammatory variables, and antioxidant capacity in Korean patients with type 2 diabetes. The results suggest that spirulina is a promising agent as a functional food for diabetes management. PMID- 20016734 TI - Nutritional status and the role of diabetes mellitus in hemodialysis patients. AB - This study was aimed to investigate the nutritional status and the role of diabetes mellitus in hemodialysis (HD) patients. Anthropometric, biochemical, and dietary assessments for HD 110 patients (46 males and 64 females) were conducted. Mean body mass index (BMI) was 22.1 kg/m(2) and prevalence of underweight (BMI<18.5 kg/m(2)) was 12%. The hypoalbuminemia (<3.5 g/dl) was found in 15.5% of the subject, and hypocholesterolemia (<150 mg/dl) in 46.4%. About half (50.9%) patients had anemia (hemoglobin: <11.0 g/dL). High prevalence of hyperphosphatemia (66.4%) and hyperkalemia (43.5%) was also observed. More than 60 percent of subjects were below the recommended intake levels of energy (30-35 kcal/kg IBW) and protein (1.2 g/kg IBW). The proportions of subjects taking less than estimated average requirements for calcium, vitamin B(1), vitamin B(2), vitamin C, and folate were more than 50%, whereas, about 20% of the subjects were above the recommended intake of phosphorus and potassium. Diabetes mellitus was the main cause of ESRD (45.5%). The diabetic ESRD patients showed higher BMI and less HD adequacy than nondiabetic patients. Diabetic patients also showed lower HDL-cholesterol levels. Diabetic ESRD patients had less energy from fat and a greater percentage of calories from carbohydrates. In conclusion, active nutrition monitoring is needed to improve the nutritional status of HD patients. A follow-up study is needed to document a causal relation between diabetes and its impact on morbidity and mortality in ESRD patients. PMID- 20016735 TI - A study on nutrition knowledge and dietary behavior of elementary school children in Seoul. AB - The purpose of this study was to investigate the nutrition and diet related knowledge, attitude, and behavior of elementary school children in Seoul. The subjects included were 439 (male 236, female 203) elementary school children in the 4th to the 6th grades. The statistical analysis was conducted using SPSS 12.0 program. The average obesity index (OI) was 104.98 and 99.82 for male and female subjects, respectively. The average percentage of underweight, normal, overweight and obese of subjects was 33.7%, 32.8%, 12.3%, and 19.4%, respectively. The percentage of the underweight group of female subjects was higher than that of the male subjects. The percentage of the obese group of male subjects was higher than that of the female subjects. The average score of nutrition knowledge, nutrition attitude and dietary behavior was 6.8, 7.44, and 7.34, respectively. Dietary behavior of male subjects was positively correlated with parents' education levels, monthly household income and nutrition attitude. Dietary behavior of female subjects was positively correlated with monthly household income, nutrition knowledge and nutrition attitude. Dietary behavior of female subjects was positively correlated with obesity index (OI). Proper nutrition education and intervention are required for the improvement of elementary school children's nutrition knowledge, nutrition attitudes and dietary behaviors. PMID- 20016736 TI - Effects of various metal ions on the gene expression of iron exporter ferroportin 1 in J774 macrophages. AB - Macrophages play a key role in iron metabolism by recycling iron through erythrophagocytosis. Ferroportin-1 (FPN1) is a transporter protein that is known to mediate iron export from macrophages. Since divalent metals often interact with iron metabolism, we examined if divalent metals could regulate the expression of FPN1 in macrophages. J774 macrophage cells were treated with copper, manganese, zinc, or cobalt at 10, 50, or 100 microM for 16 to 24 h. Then, FPN1 mRNA and protein levels were determined by quantitative real-time PCR and Western blot analyses, respectively. In addition, effects of divalent metals on FPN1 promoter activity were examined by luciferase reporter assays. Results showed that copper significantly increased FPN1 mRNA levels in a dose-dependent manner. The copper-induced expression of FPN1 mRNA was associated with a corresponding increase in FPN1 protein levels. Also, copper directly stimulated the activity of FPN1 promoter-driven reporter construct. In contrast, manganese and zinc had no effect on the FPN1 gene expression in J774 cells. Interestingly, cobalt treatment in J774 cells decreased FPN1 protein levels without affecting FPN1 mRNA levels. In conclusion, our study results demonstrate that divalent metals differentially regulate FPN1 expression in macrophages and indicate a potential interaction of divalent metals with the FPN1-mediated iron export in macrophages. PMID- 20016737 TI - Kaempferol induced the apoptosis via cell cycle arrest in human breast cancer MDA MB-453 cells. AB - The aim of present study was to investigate the effects of kaempferol on cellular proliferation and cell cycle arrest and explore the mechanism for these effects in human breast carcinoma MDA-MB-453 cells. Cells were treated with kaempferol at various concentrations (ranging from 1 to 200 microM) for 24 and 48 hrs. Kaempferol significantly inhibited cancer cell growth in cells exposed to 50 and 10 microM of kaempferol and incubated for 24 and 48 hrs, respectively. Exposure to kaempferol resulted in cell cycle arrest at the G2/M phase. Of the G2/M-phase related proteins, kaempferol down-regulated CDK1 and cyclin A and B in cells exposed to kaempferol. In addition, small DNA fragments at the sub-G0 phase were increased by up to 23.12 and 31.90% at 10 and 50 microM incubated for 24 and 48 hrs, respectively. The kaempferol-induced apoptosis was associated with the up regulation of p53. In addition, the phosphorylation of p53 at the Ser-15 residue was observed with kaempferol. Kaempferol inhibits cell proliferation by disrupting the cell cycle, which is strongly associated with the induction of arrest at G2/M phase and may induce apoptosis via p53 phosphorylation in human breast carcinoma MDA-MB-453 cells. PMID- 20016739 TI - Predictors of workplace absenteeism in cancer care workers. PMID- 20016738 TI - Isomer specificity of conjugated linoleic acid (CLA): 9E,11E-CLA. AB - Conjugated linoleic acids (CLA) were identified in 1980's, since then it has been intensively studied due to its various beneficial health effects such as anti inflammatory, anti-atherogenic, anti-carcinogenic and anti-diabetic/obesity effects. Isomer specificity of a number of CLA isomers, especially predominant isomer 9Z,11E- and 10E,12Z-CLA, is now recognized. However, the less prevalent CLA isomers have not been well characterized. Recently, studies have reported the distinctively different effects of 9E,11E-CLA in colon cancer cells, endothelial cells, and macrophage cells compared to the rest of CLA isomers. In this review, various effects of CLAs, especially anti-inflammatory and anti-atherogenic effects, will be discussed with focusing on the isomer-specific effects and potential mechanism of action of CLA. At last, recent studies about 9E,11E-CLA in in vitro and animal models will be discussed. PMID- 20016740 TI - Bogarting that joint might decrease oral HPV among cannabis users. AB - Human papilloma virus (HPV) has many known strains, two of the most well studied perhaps being the high-risk types 16 and 18. These strains have attracted more interest because they are known to disrupt tumour-suppressor genes that control the cell cycle, rendering those genes less effective at keeping cell division in check.Within the last decade, an increase in oral hpv-linked cancers of the throat and tongue has been attributed to exposure and contraction of hpv through oral sex, most notably in younger people. An understudied and arguably equal contributor to oral hpv infection might be indirect contact with an infected person. Presented here is a brief but important perspective on the relationship between cannabis use and oral cancer. The development of oral cancer is not a result of smoking cannabis per se; rather, it is hypothesized to be a result of contracting hpv through various forms of sharing and passing joints and other smoking apparatuses. Therefore, it is hypothesized that bogarting (and not passing) joints might decrease oral hpv among cannabis smokers. Future research should therefore investigate the prevalence of oral hpv in cannabis smokers to better understand its epidemiology. PMID- 20016741 TI - Relative dose intensity delivered to patients with early breast cancer: Canadian experience. AB - Adjuvant chemotherapy for early breast cancer improves disease-free and overall survival in pre- and postmenopausal women. The importance of maintaining relative dose intensity (RDI) is well-known; however, little information is available from routine clinical practice regarding how well dose intensity is maintained with modern chemotherapy regimens.In a retrospective review of patients undergoing chemotherapy for early breast cancer at a single institution in Canada from January 2006 to November 2007, a total of 263 patients received one of the following regimens:AC-T [doxorubicin (Adriamycin: Pharmacia, Kalamazoo, MI, U.S.A.)-cyclophosphamide, paclitaxel (Taxol: Bristol-Myers Squibb, Princeton, NJ, U.S.A.)]FEC-100 (5-fluorouracil-epirubicin-cyclophosphamide)FEC-D (5-fluorouracil epirubicin-cyclophosphamide, docetaxel)Overall, only 14.4% of patients had a RDI less than 85%. Dose delay or reduction (or both) occurred in 46%, 37%, and 20% of patients receiving fec-100, ac-t, and fec-d respectively. Optimal RDI was delivered to 96%, 95%, and 70.7% of patients for ac-t, fec-d and fec-100 regimens respectively. Patients over 65 years of age accounted for 14% of the total cohort and were more likely to receive a suboptimal RDI than were patients younger than 65 years of age (35% vs. 6.6%).Optimal chemotherapy RDI (>85%) for early breast cancer can be achieved at an academic cancer centre. This goal is less often accomplished in elderly patients, and thus a proactive approach is required for managing toxicity in that population. PMID- 20016742 TI - Generalizability of toxicity data from oncology clinical trials to clinical practice: toxicity of irinotecan-based regimens in patients with metastatic colorectal cancer. AB - BACKGROUND: The relevance of oncology trial results to clinical practice depends on whether the trial participants are similar to the actual population of patients receiving treatment for the malignancy and whether the patients are treated similarly in both circumstances. Chemotherapy treatments may be more toxic in patients of advanced age and poor performance status-patients typically excluded from clinical trials. METHODS: In a retrospective chart review that included all non-trial patients with metastatic colorectal cancer treated with irinotecan-based chemotherapy from January 2004 to September 2006 at our institution, we quantified and subsequently compared the toxicity rates of the irinotecan regimens in clinical practice with published toxicity rates from corresponding phase iii clinical trials. The primary endpoint was the incidence of grades 3 and 4 diarrhea. RESULTS: The study included 203 patients, and the irinotecan regimens considered included FOLFIRI [irinotecan, leucovorin, 5 fluorouracil (5fu)],IFL (bevacizumab, irinotecan, 5FU, leucovorin),XELIRI (capecitabine, 3-weekly irinotecan), andirinotecan monotherapy. The rates of grades 3 and 4 diarrhea for FOLFIRI, IFL, XELIRI, and irinotecan monotherapy in clinical practice were 10%, 15%, 17%, and 21% as compared with 10%, 23%, 20%, and 31% respectively in clinical trials. When only patients meeting trial performance status and age criteria were analyzed, the rates of grades 3 and 4 diarrhea by regimen were 11%, 20%, 19%, and 26% respectively. CONCLUSIONS: Overall, the toxicity rates for FOLFIRI and irinotecan monotherapy in non-trial patients were not statistically different from the rates quoted in published clinical trials. PMID- 20016743 TI - Impact of radiation avoidance on survival and neurocognitive outcome in infant medulloblastoma. AB - PURPOSE: Concerns about radiotherapy-related neurocognitive sequelae in young children have led to deferral or avoidance of radiation in contemporary treatment for this fragile group of patients. We compared survival and neurocognitive outcome in two groups of infants with medulloblastoma who received adjuvant conventional craniospinal irradiation (CSI) or reduced or no radiotherapy during an era of change in the philosophy of infant medulloblastoma treatment. PATIENTS AND METHODS: From 1985 to 2007, 29 patients 3 years of age or younger were diagnosed and treated with curative intent in our institution. Children treated before 1994 received adjuvant radiation with chemotherapy; subsequently, radiation was prescribed essentially for disease progression or relapse. RESULTS: Median age at diagnosis was 24 months (range: 1-36 months); 15 patients (52%) presented with metastatic disease at diagnosis. As part of initial treatment, 8 children received adjuvant radiotherapy with chemotherapy, and 21 children received postoperative chemotherapy only. Five children treated with chemotherapy alone are in prolonged remission. The 5-year event-free and overall survivals were 35.9% +/- 9.8% and 50.2% +/- 9.6% respectively. Extent of resection, metastatic status, and desmoplastic histology were not found to be significant prognostic factors. On serial neurocognitive evaluations, patients treated with chemotherapy with or without reduced radiotherapy demonstrated improvement of intellectual function over time. Patients treated with conventional csi exhibited significantly lower intelligence quotient scores and academic performance, with the exception of receptive vocabulary. CONCLUSIONS: Avoidance of conventional CSI in treatment of very young children with medulloblastoma appears to be associated with a preserved neurocognitive profile. Neurocognitive evaluation should be integrated into the primary objectives of future infant protocols. PMID- 20016744 TI - Cancer diagnostic assessment programs: standards for the organization of care in Ontario. AB - BACKGROUND: Improving access to better, more efficient, and rapid cancer diagnosis is a necessary component of a high-quality cancer system. How diagnostic services ought to be organized, structured, and evaluated is less understood and studied. Our objective was to address this gap. METHODS: As a quality initiative of Cancer Care Ontario's Program in Evidence-Based Care, the Diagnostic Assessment Standards Panel, with representation from clinical oncology experts, institutional and clinical administrative leaders, health service researchers, and methodologists, conducted a systematic review and a targeted environmental scan of the unpublished literature. Standards were developed based on expert consensus opinion informed by the identified evidence. Through external review, clinicians and administrators across Ontario were given the opportunity to provide feedback. RESULTS: The body of evidence consists of thirty-five published studies and fifteen unpublished guidance documents. The evidence and consensus opinion consistently favoured an organized, centralized system with multidisciplinary team membership as the optimal approach for the delivery of diagnostic cancer assessment services. Independent external stakeholders agreed (with higher mean values, maximum 5, indicating stronger agreement) that DAP standards are needed (mean: 4.6), that standards should be formally approved (mean: 4.3), and importantly, that standards reflect an effective approach that will lead to quality improvements in the cancer system (mean: 4.5) and in patient care (mean: 4.3). INTERPRETATION: Based on the best available evidence, standards for the organization of DAPS are offered. There is clear need to integrate formal and comprehensive evaluation strategies with the implementation of the standards to advance this field. PMID- 20016745 TI - Clinician-patient communication: evidence-based recommendations to guide practice in cancer. AB - GOALS OF WORK: To develop recommendations for effective communication between cancer health care providers and patients based on a systematic review of methods of clinician-patient communication that may affect patient outcomes associated with distress at critical points in the course of cancer care. MATERIALS AND METHODS: A systematic review of the literature was conducted, and evidence-based recommendations were formulated to guide clinician-patient communication in cancer care. A formal external review was conducted to validate the relevance of these recommendations. MAIN RESULTS: Recommendations for communication in cancer care are presented, based on guidelines from the Australian National Breast Cancer Centre and the Australian National Cancer Control Initiative,an updated systematic review of the research evidence, and a consensus by the Clinician Patient Communications Working Panel of the Program in Evidence-Based Care of Cancer Care Ontario. The recommendations were sent to 110 Ontario practitioners for external review: 33 responded (30% response rate). Most of these respondents (87%) agreed with the draft recommendations and approved of their use as a practice guideline (90%). A condensed version of the recommendations, including 10 key points, was also created. CONCLUSIONS: There is evidence to support general clinician-patient communication approaches, although the preferences of cancer patients regarding such communication exhibit individual and cultural variability. Recommendations are provided, based on evidence, the consensus of an expert panel, and feedback from a survey of external practitioners. Evidence evaluating the role of decision aids and strategies to facilitate better communication is inconsistent, although such tools may be of value for some patients. PMID- 20016746 TI - Colorectal Cancer Association of Canada consensus meeting: raising the standards of care for early-stage rectal cancer. AB - The purpose of the meeting reported here was to develop a set of national evidence-based standards for assessing and managing patients with potentially resectable rectal cancer. This report represents the consensus of the multidisciplinary group of Canadian rectal cancer experts attending that meeting. PMID- 20016747 TI - Partial cancerous changes eventually developing into superficial spreading cancer over 18 months. AB - In this report, we describe a patient presenting with the superficial spreading type of early gastric cancer (EGC) accompanied by cancerous ulcers. Disease progression and treatment outcome are discussed. After symptoms persisted for more than 1 year, the patient underwent total gastrectomy with D2 lymph node dissection. The patient was diagnosed with superficial spreading cancer (SSC), accompanied by an extensive iic lesions. The progression of this patient suggests that the co-occurrence of cancerous ulcers may contribute to EGC development to some extent. As is known, EGC often develops into advanced gastric cancer with time. However, in our case, we observed a process during which partial cancerous changes developed into ssc over 18 months. Superficial spreading cancer should be considered an EGC variant, which may have the ability to spread superficially along the stomach wall without invading the muscularis propria. But we speculate that, if gene expression changes for some reason, the malignant SSC cells may acquire the ability to grow deeply into the stomach wall. Eventually, Borrmann type IV gastric cancer may develop. PMID- 20016748 TI - Identification of Anoectochilus based on rDNA ITS sequences alignment and SELDI TOF-MS. AB - The internal transcribed spacer (ITS) sequences alignment and proteomic difference of Anoectochilus interspecies have been studied by means of ITS molecular identification and surface enhanced laser desorption ionization time of flight mass spectrography. Results showed that variety certification on Anoectochilus by ITS sequences can not determine species, and there is proteomic difference among Anoectochilus interspecies. Moreover, proteomic finger printings of five Anoectochilus species have been established for identifying species, and genetic relationships of five species within Anoectochilus have been deduced according to proteomic differences among five species. PMID- 20016749 TI - Initiation and persistence to statin treatment in patients with diabetes receiving glucose-lowering medications 1997- 2006. AB - AIMS: Since 2001 guidelines recommend statin treatment in most patients with diabetes. We investigated secular changes in initiation and persistence to statin treatment during a 10-year period in a nationwide cohort of patients initiating glucose-lowering medication (GLM). METHODS: All Danish citizens 30 years and older who claimed prescriptions of GLM between 1997 and 2006 were identified from nationwide registers of drug dispensing from pharmacies and hospitalizations, and followed until 2006. Statin treatment was registered if a prescription was claimed during the period. By logistic regression we analyzed factors related to initiation and persistence to statin treatment. RESULTS: In total 128,106 patients were included. In 1997 only 7% of the patients receiving GLM claimed statins within the first year after GLM initiation. Despite increasing statin prescriptions the following years, only 62% were using statins at the end of follow up. The chance of ever receiving statins was lowest if not initiated within 180-days following the first purchase of GLM (OR 0.75, 95% CI 0.74-0.76). A previous myocardial infarction was associated with increased statin treatment (OR 4.51; 95% CI 4.31 - 4.71), while low income was associated with lower use of statins (OR 0.68; 95%CI 0.66-0.72). Between 75-85 % of the patients who initiated statins treatment were persistent to treatment by 2007. CONCLUSIONS: In spite of increasing use of statins in diabetes patients over time, many patients remain untreated. Early initiation of statin treatment in diabetic patients and focus on patients with low socioeconomic status is needed to give long-term benefits. PMID- 20016750 TI - Application of One Sided t-tests and a Generalized Experiment Wise Error Rate to High-Density Oligonucleotide Microarray Experiments: An Example Using Arabidopsis. AB - MOTIVATION: A formidable challenge in the analysis of microarray data is the identification of those genes that exhibit differential expression. The objectives of this research were to examine the utility of simple ANOVA, one sided t tests, natural log transformation, and a generalized experiment wise error rate methodology for analysis of such experiments. As a test case, we analyzed a Affymetrix GeneChip microarray experiment designed to test for the effect of a CHD3 chromatin remodeling factor, PICKLE, and an inhibitor of the plant hormone gibberellin (GA), on the expression of 8256 Arabidopsis thaliana genes. RESULTS: The GFWER(k) is defined as the probability of rejecting k or more true null hypothesis at a given p level. Computing probabilities by GFWER(k) was shown to be simple to apply and, depending on the value of k, can greatly increase power. A k value as small as 2 or 3 was concluded to be adequate for large or small experiments respectively. A one sided t-test along with GFWER(2)=.05 identified 43 genes as exhibiting PICKLE-dependent expression. Expression of all 43 genes was re-examined by qRT-PCR, of which 36 (83.7%) were confirmed to exhibit PICKLE-dependent expression. PMID- 20016751 TI - Prognostic Significance and Clinicopathological Associations of COX-2 SNP in Patients with Nonsmall Cell Lung Cancer. AB - Background. To further improve the screening, diagnosis, and therapy of patients with nonsmall cell lung cancer (NSCLC) additional diagnostic tools are urgently needed. Gene expression of Cyclooxygenase-2 (COX-2) has been linked to prognosis in patients with NSCLC. The role of the COX-2 926G>C Single Nucleotide Polymorphism (SNP) in patients with NSCLC remains unclear. The aim of this study was to investigate the potential of the COX-2 926G>C SNP as a molecular marker in this disease. Methods. COX-2 926G>C SNP was analyzed in surgically resected tumor tissue of 85 patients with NSCLC using a PCR-based RFLP technique. Results. The COX-2 926G>C SNP genotypes were detected with the following frequencies: GG n = 62 (73%), GC n = 20 (23%), CC n = 3 (4%). There were no associations between COX 2 SNP genotype and histology, grading or gender detectable. COX-2 SNP was significantly associated with tumor stage (P = .032) and lymph node status (P = .016, Chi-square test). With a median followup of 85.9 months, the median survival was 59.7 months. There were no associations seen between the COX-2 SNP genotype and patients prognosis. Conclusions. The COX-2 926G>C SNP is detectable at a high frequency in patients with NSCLC. The COX-2 926G>C SNP genotype is not a prognostic molecular marker in this disease. However, patients with the GC or CC genotype seem more susceptible to lymph node metastases and higher tumor stage than patients with the GG genotype. The results suggest COX-2 926G>C SNP as a molecular marker for lymph node involvement in this disease. PMID- 20016752 TI - Significance of Circulating Tumor Cells Detected by the CellSearch System in Patients with Metastatic Breast Colorectal and Prostate Cancer. AB - The increasing number of treatment options for patients with metastatic carcinomas has created a concomitant need for new methods to monitor their use. Ideally, these modalities would be noninvasive, be independent of treatment, and provide quantitative real-time analysis of tumor activity in a variety of carcinomas. Assessment of circulating tumor cells (CTCs) shed into the blood during metastasis may satisfy this need. We developed the CellSearch System to enumerate CTC from 7.5 mL of venous blood. In this review we compare the outcomes from three prospective multicenter studies investigating the use of CTC to monitor patients undergoing treatment for metastatic breast (MBC), colorectal (MCRC), or prostate cancer (MPC) and review the CTC definition used in these studies. Evaluation of CTC at anytime during the course of disease allows assessment of patient prognosis and is predictive of overall survival. PMID- 20016753 TI - A computationally efficient formal optimization of regional myocardial contractility in a sheep with left ventricular aneurysm. AB - A non-invasive method for estimating regional myocardial contractility in vivo would be of great value in the design and evaluation of new surgical and medical strategies to treat and/or prevent infarction-induced heart failure. As a first step towards developing such a method, an explicit finite element (FE) model based formal optimization of regional myocardial contractility in a sheep with left ventricular (LV) aneurysm was performed using tagged magnetic resonance (MR) images and cardiac catheterization pressures. From the tagged MR images, 3 dimensional (3D) myocardial strains, LV volumes and geometry for the animal specific 3D FE model of the LV were calculated, while the LV pressures provided physiological loading conditions. Active material parameters (T(max_B) and T(max_R)) in the non-infarcted myocardium adjacent to the aneurysm (borderzone) and in myocardium remote from the aneurysm were estimated by minimizing the errors between FE model-predicted and measured systolic strains and LV volumes using the successive response surface method for optimization. The significant depression in optimized T(max_B) relative to T(max_R) was confirmed by direct ex vivo force measurements from skinned fiber preparations. The optimized values of T(max_B) and T(max_R) were not overly sensitive to the passive material parameters specified. The computation time of less than 5 hours associated with our proposed method for estimating regional myocardial contractility in vivo makes it a potentially very useful clinical tool. PMID- 20016755 TI - Oxidative reactions of tetrahydrobenzimidazole derivatives with N sulfonyloxaziridines. AB - An investigation of the utility of N-sulfonyloxaziridines to effect the oxidative rearrangement of tetrahydrobenzimidazoles to spiro fused 5-imidazolones is reported. In addition to the anticipated rearrangement manifold, it was found that 2-amino substituted derivatives afford products resulting from rearrangement, or alternatively from addition of methanol or water depending on the nature of the N-substituents and reaction conditions. PMID- 20016754 TI - Hypophosphatemia, hyperphosphaturia, and bisphosphonate treatment are associated with survival beyond infancy in generalized arterial calcification of infancy. AB - BACKGROUND: Generalized arterial calcification of infancy has been reported to be frequently lethal, and the efficiency of any therapy, including bisphosphonates, is unknown. A phosphate-poor diet markedly increases survival of NPP1 null mice, a model of generalized arterial calcification of infancy. METHODS AND RESULTS: We performed a multicenter genetic study and retrospective observational analysis of 55 subjects affected by generalized arterial calcification of infancy to identify prognostic factors. Nineteen (34%) patients survived the critical period of infancy. In all 8 surviving patients tested, hypophosphatemia due to reduced renal tubular phosphate reabsorption developed during childhood. Eleven of 17 (65%) patients treated with bisphosphonates survived. Of 26 patients who survived their first day of life and were not treated with bisphosphonates only 8 (31%) patients survived beyond infancy. Forty different homozygous or compound heterozygous mutations, including 16 novel mutations in ENPP1, were found in 41 (75%) of the 55 patients. Twenty-nine (71%) of these 41 patients died in infancy (median, 30 days). Seven of the 14 (50%) patients without ENPP1 mutations died in infancy (median, 9 days). When present on both alleles, the mutation p.P305T was associated with death in infancy in all 5 cases; otherwise, no clear genotype phenotype correlation was seen. CONCLUSION: ENPP1 coding region mutations are associated with generalized arterial calcification of infancy in approximately 75% of subjects. Except for the p.P305T mutation, which was universally lethal when present on both alleles, the identified ENPP1 mutations per se have no discernable effect on survival. However, survival seems to be associated with hypophosphatemia linked with hyperphosphaturia and also with bisphosphonate treatment. PMID- 20016757 TI - Bayesian Analysis of Multivariate Latent Curve Models With Nonlinear Longitudinal Latent Effects. AB - In longitudinal studies, investigators often measure multiple variables at multiple time points and are interested in investigating individual differences in patterns of change on those variables. Furthermore, in behavioral, social, psychological, and medical research, investigators often deal with latent variables that cannot be observed directly and should be measured by 2 or more manifest variables. Longitudinal latent variables occur when the corresponding manifest variables are measured at multiple time points. Our primary interests are in studying the dynamic change of longitudinal latent variables and exploring the possible interactive effect among the latent variables.Much of the existing research in longitudinal studies focuses on studying change in a single observed variable at different time points. In this article, we propose a novel latent curve model (LCM) for studying the dynamic change of multivariate manifest and latent variables and their linear and interaction relationships. The proposed LCM has the following useful features: First, it can handle multivariate variables for exploring the dynamic change of their relationships, whereas conventional LCMs usually consider change in a univariate variable. Second, it accommodates both first- and second-order latent variables and their interactions to explore how changes in latent attributes interact to produce a joint effect on the growth of an outcome variable. Third, it accommodates both continuous and ordered categorical data, and missing data. PMID- 20016756 TI - FRET and mechanobiology. AB - Since the development of green fluorescent protein (GFP) and other fluorescent proteins (FPs) with distinct colors, genetically-encoded probes and biosensors have been widely applied to visualize the molecular localization and activities in live cells. In particular, biosensors based on fluorescence resonance energy transfer (FRET) have significantly advanced our understanding of the dynamic molecular hierarchy at subcellular levels. These biosensors have also been extensively applied in recent years to study how cells perceive the mechanical environment and transmit it into intracellular molecular signals (i.e. mechanotransduction). In this review, we will first provide a brief introduction of the recent development of FPs. Different FRET biosensors based on FPs will then be described. The last part of the review will be dedicated to the introduction of examples applying FRET biosensors to visualize mechanotransduction in live cells. In summary, the integration of FRET technology and the different cutting-edge mechanical stimulation systems can provide powerful tools to allow the elucidation of the mechanisms regulating mechanobiology at cellular and molecular levels in normal and pathophysiological conditions. PMID- 20016758 TI - Mother and Adolescent Reports of Interparental Discord among Parents of Adolescents with and without Attention-Deficit Hyperactivity Disorder. AB - Evidence is scarce regarding the prevalence of interparental discord in families of adolescents with attention-deficit hyperactivity disorder (ADHD). Using data collected from adolescents with childhood ADHD and comorbid oppositional-defiant disorder (ODD; n = 46) or conduct disorder (CD; n =23), with childhood ADHD-only (n = 26), and without ADHD (n = 88), and their mothers, maternal and adolescent reports of interparental discord were compared. Adolescents with ADHD+CD reported witnessing more frequent and unresolved interparental conflict than adolescents without ADHD and with ADHD-Only. Adolescents with ADHD+CD also indicated more frequent conflict than adolescents with ADHD+ODD. However, differences in conflict resolution were nonsignificant when household income was covaried and maternal ratings of interparental discord did not differ across groups. Findings highlight the potential utility of adolescents with ADHD as informants of interparental relationship quality. PMID- 20016759 TI - Patterns of Competence and Adjustment Among Adolescents from Authoritative, Authoritarian, Indulgent, and Neglectful Homes: A Replication in a Sample of Serious Juvenile Offenders. AB - The correlates of authoritative, authoritarian, indulgent, and neglectful parenting were examined within a sample of 1,355 14- to 18-year-olds adjudicated of serious criminal offenses. The sample is composed primarily of poor, ethnic minority youth living in impoverished urban neighborhoods. As has been found in community samples, juvenile offenders who describe their parents as authoritative are more psychosocially mature, more academically competent, less prone to internalized distress, and less prone to externalizing problems than their peers,whereas those who describe their parents as neglectful are less mature, less competent, and more troubled. Juvenile offenders who characterize their parents as either authoritarian or indulgent typically score somewhere between the two extremes, although those from authoritarian homes are consistently better functioning than those from indulgent homes. These patterns did not vary as a function of adolescents' ethnicity or gender. PMID- 20016760 TI - WOUND-HEALING PROPERTIES OF TRANSFORMING GROWTH FACTOR beta (TGF-beta) INDUCIBLE EARLY GENE 1 (TIEG1) KNOCKOUT MICE. AB - Transforming growth factor beta (TGF-beta) has a broad effect on wound healing, but many questions remain about the regulation of TGF-beta during the healing process. TGF-beta inducible early gene 1 (TIEG1) is a primary response gene for TGF-beta that controls the activities of the TGF-beta/Smad pathway, the primary TGF-beta signaling pathway. The purpose of this study was to investigate the role of TIEG1 in cutaneous wound healing using TIEG1 knockout mice. The wound healing in TIEG1 knockout mice and wild-type controls was evaluated by wound breaking strength, Western blot, and histology at postoperative days 3, 7, and 14. Although re-epithelialization of both groups was similarly complete at day 7, the TIEG1 knockout mice had a significantly lower wound breaking strength than the controls at postoperative day 14. These results suggest that TIEG1 expression may be an important factor involved in the initiation and support of normal cutaneous wound healing. PMID- 20016761 TI - Making the most of Papua New Guinea's biodiversity: Establishment of an integrated set of programs that link botanical survey with pharmacological assessment in "The Land of the Unexpected" AB - An integrated and coordinated set of programs has been established to meet ICBG goals in Papua New Guinea (PNG). Here we give an overview of the PNG ICBG and focus on the key elements and major steps taken to establish a program necessary for the pharmacological assessment of botanicals and traditional medicines in PNG and, by extrapolation, in other developing countries. PMID- 20016763 TI - Synthesis and Characterization of Ruthenium(II) Phenanthroline Complexes Containing Quaternary Amine Substituents. AB - A series of mixed ligand complexes of ruthenium(II) complexes containing 5 methylphenanthroline and trimethylamino-5-methylphenanthroline have been synthesized to investigate the impact of the quaternary amine on the photophysical properties. Thermal stability studies indicate that the quaternary amine group is stable with respect to hydrolysis. Mass spectral analysis of the complexes revealed only fragments consistent with homolytic cleavage of the amines and no parent ions were observed. Both electrochemical and photophysical investigations indicate that the quaternary amine has little or no impact on the properties of the complex when compared to the complexes lacking the amine. PMID- 20016762 TI - Sox2 and Oct-3/4: a versatile pair of master regulators that orchestrate the self renewal and pluripotency of embryonic stem cells. AB - During the past 10 years, remarkable progress has been made in understanding the transcriptional mechanisms that control the biology of stem cells. Given the importance of stem cells in development, regenerative medicine, and cancer, it is no surprise that the pace of discovery continues to accelerate--paradigm-shifting models proposed only a few years ago are quickly giving way to even more sophisticated models of regulation. This review summarizes some of the major advances made in delineating the roles of two transcription factors, Sox2 and Oct 3/4, in stem cell biology. Additionally, unanswered questions related to their mechanisms of action are discussed. When viewed together, it is evident that Sox2 and Oct-3/4 exhibit the major properties expected of master regulators. They are each essential for mammalian development, they help regulate the transcription of other genes that are essential for development, and they influence their own transcription by both positive and negative feedback loops. Moreover, small changes in the levels of either Sox2 or Oct-3/4 trigger the differentiation of embryonic stem (ES) cells. Thus, each functions as a molecular rheostat to control the self-renewal and pluripotency of ES cells. Overall, understanding how Sox2 and Oct-3/4 function mechanistically will not only provide important insights into stem cells in general, but should also have a significant impact on our understanding of induced pluripotent stem (iPS) cells and, hence, the emerging field of regenerative medicine. PMID- 20016764 TI - COMMUNITY VIOLENCE AND EXTERNALIZING PROBLEMS: MODERATING EFFECTS OF RACE AND RELIGIOSITY IN EMERGING ADULTHOOD. AB - The present study examined racial differences in the relationship between exposure to community violence and public and private religiosity in predicting externalizing problems among at-risk emerging adults. Participants were 178 African American and 163 European American emerging adults at risk for exposure to community violence. Exposure to community violence related to more externalizing problems. Greater public religious affiliation buffered the relationship between community violence and substance abuse, for both African American and European emerging adults. In addition, more privately religious African American emerging adults engaged in less deviant behavior when exposed to higher levels of community violence. European Americans were not protected by private religiosity. PMID- 20016765 TI - Micromanaging Palate Development. AB - Development of the facial skeleton is one of the most intriguing and intricate events that occur during human development. Most of the bone, cartilage and connective tissue that compose the face and neck arise from a class of cells, referred to as neural crest cells, which are initially located at some distance from the facial primordium. A complex set of events regulated by specific gene products direct the formation, migration and differentiation of these cells, leading to what is viewed as "prototypical" adult facial features. These basic developmental processes are recapitulated during the formation of the palate, termed palatogenesis. In this review, we summarize the basic embryology leading to palate formation, discuss mechanisms that can lead to palatal dysmorphologies and highlight a new interaction that has recently been demonstrated to play a role in palate development. This interaction, involving small non-coding RNAs referred to as microRNAs, not only establishes a new level of regulation to cellular development, but may also serve as attractive targets for future efforts directed at clinical treatment of birth defect syndromes. PMID- 20016766 TI - Optoacoustic generation of high frequency sound for 3-D ultrasonic imaging in medicine. AB - The most common form of optoacoustic generation is thermoelasticity. Thermoelastic transduction is easy to implement and can be very broadband. However, its major drawback has always been poor conversion efficiency when a metallic film is used as the transducer. We have investigated two alternate structures for high efficiency, one based on a thin polymer film and the other using a two-dimensional nanostructure. PMID- 20016767 TI - A Case Study in Pharmacologic Colon Imaging Using Principal Curves in Single Photon Emission Computed Tomography. AB - In this manuscript we are concerned with functional imaging of the colon to assess the kinetics of microbicide lubricants. The overarching goal is to understand the distribution of the lubricants in the colon. Such information is crucial for understanding the potential impact of the microbicide on HIV viral transmission. The experiment was conducted by imaging a radiolabeled lubricant distributed in the subject's colon. The tracer imaging was conducted via single photon emission computed tomography (SPECT), a non-invasive, in-vivo functional imaging technique. We develop a novel principal curve algorithm to construct a three dimensional curve through the colon images. The developed algorithm is tested and debugged on several difficult two-dimensional images of familiar curves where the original principal curve algorithm does not apply. The final curve fit to the colon data is compared with experimental sigmoidoscope collection. PMID- 20016768 TI - DIFFEOMORPHIC SURFACE FLOWS: A NOVEL METHOD OF SURFACE EVOLUTION. AB - We describe a new class of surface flows, diffeomorphic surface flows, induced by restricting diffeomorphic flows of the ambient Euclidean space to a surface. Different from classical surface PDE flows such as mean curvature flow, diffeomorphic surface flows are solutions of integro-differential equations in a group of diffeomorphisms. They have the potential advantage of being both topology-invariant and singularity free, which can be useful in computational anatomy and computer graphics. We first derive the Euler-Lagrange equation of the elastic energy for general diffeomorphic surface flows, which can be regarded as a smoothed version of the corresponding classical surface flows. Then we focus on diffeomorphic mean curvature flow. We prove the short-time existence and uniqueness of the flow, and study the long-time existence of the flow for surfaces of revolution. We present numerical experiments on synthetic and cortical surfaces from neuroimaging studies in schizophrenia and auditory disorders. Finally we discuss unresolved issues and potential applications. PMID- 20016769 TI - Meeting the Challenges of Neuroimaging Genetics. AB - As research encompassing neuroimaging and genetics gains momentum, extraordinary information will be uncovered on the genetic architecture of the human brain. However, there are significant challenges to be addressed first. Not the least of these challenges is to accomplish the sample size necessary to detect subtle genetic influences on the morphometry and function of the healthy brain. Aside from sample size, image acquisition and analysis methods need to be refined in order to ensure optimum sensitivity to genetic and complementary environmental influences. Then there is the vexing issue of interpreting the resulting data. We describe how researchers from the east coast of Australia and the west coast of America have embarked upon a collaboration to meet these challenges using data currently being collected from a large-scale twin study, and offer some opinions about future directions in the field. PMID- 20016770 TI - History of Violence as a Predictor of HIV Risk among Multi-Ethnic, Urban Youth in the Southwest. AB - This community-based exploratory study examined the effects of a history of violence, ethnic identification, and acculturation status on HIV risk among a majority Latino sample of youth living in a large metropolitan area of the Southwest in the United States. The participants reported high rates of violence and attitudes that put them at risk for HIV/AIDS infection. They participated in 1 of 2 prevention interventions offered by a local non-governmental organization. The first intervention was tailored for adjudicated youth (N=49) who were either institutionalized or were returning to the community after involvement with the criminal justice system. The second intervention targeted youth (N=32) who were homeless/runaway and/or self-identified as gay, lesbian, bisexual, or transgender (GLBT). T-tests and linear regression were used to determine the differences between youth reporting a history of violence by type of perpetrator, its relationship with HIV risk, and the role of ethnic identification and acculturation status as potential protective factors. Violence by a family member was the most common type of violence reported, with a history of violence positively related to HIV risk. Ethnic identification and linguistic acculturation had a protective effect against HIV risk among the homeless and GLBT youth but not among the adjudicated youth. PMID- 20016771 TI - Relapse to drug seeking following prolonged abstinence: the role of environmental stimuli. AB - Successful treatment of drug addiction must involve relapse prevention informed by our understanding of the neurobiological bases of drug relapse. In humans, exposure to drug-associated environmental stimuli can elicit drug craving and relapse. Because exposure to drug-paired stimuli similarly induces drug-seeking behavior in laboratory animals, several animal models of drug relapse have been developed. Here, we review animal models of cue-induced drug relapse and critically evaluate their validity and utility in addressing human relapse behaviors. PMID- 20016772 TI - Reward Comparison: The Achilles' heel and hope for addiction. AB - In the words of the late Charles Flaherty, reward comparison is commonplace. Rats and man, it appears, compare all rewards and this capacity likely contributes to our ability to select the most appropriate reward/behavior (food, water, salt, sex), at the most ideal level (e.g., a certain sweetness), at any given time. A second advantage of our predisposition for reward comparison is that the availability of rich alternative rewards can protect against our becoming addicted to any single reward/behavior. Thus, the potential protective effects of natural rewards/enrichment are addressed. Despite this, behavior can become inflexible when, through the development of addiction, stress, drug, or cues elicit craving, withdrawal, and ultimately, drug-seeking. Drug-seeking corresponds with a 'window of inopportunity', when even potent natural rewards have little or no impact on behavior. During this time, there is a unitary solution to the need state, and that solution is drug. The present animal model explores this 'window of inopportunity' when natural rewards are devalued and drug-seeking is engaged and considers a mode of possible intervention. PMID- 20016774 TI - Importance of basophil activation testing in insect venom allergy. AB - BACKGROUND: Venom immunotherapy (VIT) is the only effective treatment for prevention of serious allergic reactions to bee and wasp stings in sensitized individuals. However, there are still many questions and controversies regarding immunotherapy, like selection of the appropriate allergen, safety and long term efficacy. METHODS: Literature review was performed to address the role of basophil activation test (BAT) in diagnosis of venom allergy. RESULTS: In patients with positive skin tests or specific IgE to both honeybee and wasp venom, IgE inhibition test can identify sensitizing allergen only in around 15% and basophil activation test increases the identification rate to around one third of double positive patients. BAT is also diagnostic in majority of patients with systemic reactions after insect stings and no detectable IgE. High basophil sensitivity to allergen is associated with a risk of side effects during VIT. Persistence of high basophil sensitivity also predicts a treatment failure of VIT. CONCLUSION: BAT is a useful tool for better selection of allergen for immunotherapy, for identification of patients prone to side effects and patients who might be treatment failures. However, long term studies are needed to evaluate the accuracy of the test. PMID- 20016775 TI - Hodgkin's lymphoma presenting with markedly elevated IgE: a case report. AB - BACKGROUND: Markedly elevated IgE as a manifestation of a lymphoproliferative disorder has been only rarely reported. CASE PRESENTATION: We present the case of a 22 year old female referred to the adult Allergy & Clinical Immunology clinic for an extremely elevated IgE level, eventually diagnosed with Hodgkin's lymphoma. She had no history of atopy, recurrent infections, eczema or periodontal disease; stool was negative for ova & parasites. Chest X-ray revealed large bilateral anterior mediastinal masses that demonstrated prominent uptake on gallium scan. Mediastinal lymph node biopsy was consistent with Hodgkin's lymphoma, nodular sclerosing subtype, grade I/II. CONCLUSION: Although uncommon, markedly elevated IgE may be a manifestation of a malignant process, most notably both Hodgkin's and Non-Hodgkin's lymphomas. This diagnosis should be considered in evaluating an otherwise unexplained elevation of IgE. PMID- 20016773 TI - Stress and Rodent Models of Drug Addiction: Role of VTA-Accumbens-PFC-Amygdala Circuit. AB - Stress can trigger, intensify, and prolong drug consumption, as well as reinstate previously extinguished drug-taking behavior by directly impacting a neural circuit often referred to as a reward pathways. Animal models of drug abuse have been used to understand these neural circuits mediating stress-induced drug intake and relapse through examination of cellular and subcellular molecular mechanisms. Several types of intermittent stressors have been shown to induce cross-sensitization to psychomotor stimulants, enhance conditioned place preference under most conditions, increase self-administration of cocaine and amphetamine and induce reinstatement of heroin and cocaine seeking via activation of the mesocorticolimbic dopamine system. PMID- 20016776 TI - How safe are the biologicals in treating asthma and rhinitis? AB - A number of biological agents are available or being investigated for the treatment of asthma and rhinitis. The safety profiles of these biologic agents, which may modify allergic and immunological diseases, are still being elucidated. Subcutaneous allergen immunotherapy, the oldest biologic agent in current use, has the highest of frequency of the most serious and life-threatening reaction, anaphylaxis. It is also one of the only disease modifying interventions for allergic rhinitis and asthma. Efforts to seek safer and more effective allergen immunotherapy treatment have led to investigations of alternate routes of delivery and modified immunotherapy formulations. Sublingual immunotherapy appears to be associated with a lower, but not zero, risk of anaphylaxis. No fatalities have been reported to date with sublingual immunotherapy. Immunotherapy with modified formulations containing Th1 adjuvants, DNA sequences containing a CpG motif (CpG) and 3-deacylated monophospholipid A, appears to provide the benefits of subcutaneous immunotherapy with a single course of 4 to 6 preseasonal injections. There were no serious treatment-related adverse events or anaphylaxis in the clinical trials of these two immunotherapy adjuvants. Omalizumab, a monoclonal antibody against IgE, has been associated with a small risk of anaphylaxis, affecting 0.09% to 0.2% of patients. It may also be associated with a higher risk of geohelminth infection in patients at high risk for parasitic infections but it does not appear to affect the response to treatment or severity of the infection. Clinical trials with other biologic agents that have targeted IL-4/IL-13, or IL-5, have not demonstrated any definite serious treatment-related adverse events. However, these clinical trials were generally done in small populations of asthma patients, which may be too small for uncommon side effects to be identified. There is conflicting information about the safety TNF-alpha blocking agents, which have been primarily used in the treatment of rheumatoid arthritis, with serious infections, cardiovascular disease and malignancies being the most frequent serious adverse events. An unfavorable risk-benefit profile led to early discontinuation of a TNF-blocking agent in a double-blind placebo controlled of severe asthmatics. In summary, the risk of anaphylaxis and other treatment-related serious events with of all of the biological agents in this review were relatively small. However, most of the clinical trials were done in relatively small patient populations and were of relatively short duration. Long term studies in large patient populations may help clarify the risk-benefit profile of these biologic agents in the treatment of asthma. PMID- 20016777 TI - Determinants in early life for asthma development. AB - A reliable screening test in newborns for the subsequent development of bronchial asthma (BA) has not been found yet. This is mainly due to the complexity of BA, being made up by different types and underlying mechanisms. In different studies, a number of risk factors for BA have been identified. These include a positive family history of BA, passive smoking (also during pregnancy), prematurity (including pulmonary infections, RDS and BPD), early viral respiratory infections (such as RSV-bronchiolitis), male gender, early lung function abnormalities and atopic constitution. The major risk factor for persistent BA is an underlying allergic constitution. Therefore, early symptoms and markers of allergy (i.e. The Allergic March) and a positive family history for allergy should be considered as important risk factors for the development of BA. PMID- 20016778 TI - Computer modeling describes gravity-related adaptation in cell cultures. AB - Questions about the changes of biological systems in response to hostile environmental factors are important but not easy to answer. Often, the traditional description with differential equations is difficult due to the overwhelming complexity of the living systems. Another way to describe complex systems is by simulating them with phenomenological models such as the well-known evolutionary agent-based model (EABM). Here we developed an EABM to simulate cell colonies as a multi-agent system that adapts to hyper-gravity in starvation conditions. In the model, the cell's heritable characteristics are generated and transferred randomly to offspring cells. After a qualitative validation of the model at normal gravity, we simulate cellular growth in hyper-gravity conditions. The obtained data are consistent with previously confirmed theoretical and experimental findings for bacterial behavior in environmental changes, including the experimental data from the microgravity Atlantis and the Hypergravity 3000 experiments. Our results demonstrate that it is possible to utilize an EABM with realistic qualitative description to examine the effects of hypergravity and starvation on complex cellular entities. PMID- 20016779 TI - Smoking is associated with shortened airway cilia. AB - BACKGROUND: Whereas cilia damage and reduced cilia beat frequency have been implicated as causative of reduced mucociliary clearance in smokers, theoretically mucociliary clearance could also be affected by cilia length. Based on models of mucociliary clearance predicting that cilia length must exceed the 6 7 microm airway surface fluid depth to generate force in the mucus layer, we hypothesized that cilia height may be decreased in airway epithelium of normal smokers compared to nonsmokers. METHODOLOGY/PRINCIPAL FINDINGS: Cilia length in normal nonsmokers and smokers was evaluated in aldehyde-fixed, paraffin-embedded endobronchial biopsies, and air-dried and hydrated samples were brushed from human airway epithelium via fiberoptic bronchoscopy. In 28 endobronchial biopsies, healthy smoker cilia length was reduced by 15% compared to nonsmokers (p<0.05). In 39 air-dried samples of airway epithelial cells, smoker cilia length was reduced by 13% compared to nonsmokers (p<0.0001). Analysis of the length of individual, detached cilia in 27 samples showed that smoker cilia length was reduced by 9% compared to nonsmokers (p<0.05). Finally, in 16 fully hydrated, unfixed samples, smoker cilia length was reduced 7% compared to nonsmokers (p<0.05). Using genome-wide analysis of airway epithelial gene expression we identified 6 cilia-related genes whose expression levels were significantly reduced in healthy smokers compared to healthy nonsmokers. CONCLUSIONS/SIGNIFICANCE: Models predict that a reduction in cilia length would reduce mucociliary clearance, suggesting that smoking-associated shorter airway epithelial cilia play a significant role in the pathogenesis of smoking-induced lung disease. PMID- 20016780 TI - Birth weight in relation to leisure time physical activity in adolescence and adulthood: meta-analysis of results from 13 nordic cohorts. AB - BACKGROUND: Prenatal life exposures, potentially manifested as altered birth size, may influence the later risk of major chronic diseases through direct biologic effects on disease processes, but also by modifying adult behaviors such as physical activity that may influence later disease risk. METHODS/PRINCIPAL FINDINGS: We investigated the association between birth weight and leisure time physical activity (LTPA) in 43,482 adolescents and adults from 13 Nordic cohorts. Random effects meta-analyses were performed on categorical estimates from cohort , age-, sex- and birth weight specific analyses. Birth weight showed a reverse U shaped association with later LTPA; within the range of normal weight the association was negligible but weights below and above this range were associated with a lower probability of undertaking LTPA. Compared with the reference category (3.26-3.75 kg), the birth weight categories of 1.26-1.75, 1.76-2.25, 2.26-2.75, and 4.76-5.25 kg, had odds ratios of 0.67 (95% confidence interval: 0.47, 0.94), 0.72 (0.59, 0.88), 0.89 (0.79, 0.99), and 0.65 (0.50, 0.86), respectively. The shape and strength of the birth weight-LTPA association was virtually independent of sex, age, gestational age, educational level, concurrent body mass index, and smoking. CONCLUSIONS/SIGNIFICANCE: The association between birth weight and undertaking LTPA is very weak within the normal birth weight range, but both low and high birth weights are associated with a lower probability of undertaking LTPA, which hence may be a mediator between prenatal influences and later disease risk. PMID- 20016781 TI - A theoretical model for ROP localisation by auxin in Arabidopsis root hair cells. AB - BACKGROUND: Local activation of Rho GTPases is important for many functions including cell polarity, morphology, movement, and growth. Although a number of molecules affecting Rho-of-Plants small GTPase (ROP) signalling are known, it remains unclear how ROP activity becomes spatially organised. Arabidopsis root hair cells produce patches of ROP at consistent and predictable subcellular locations, where root hair growth subsequently occurs. METHODOLOGY/PRINCIPAL FINDINGS: We present a mathematical model to show how interaction of the plant hormone auxin with ROPs could spontaneously lead to localised patches of active ROP via a Turing or Turing-like mechanism. Our results suggest that correct positioning of the ROP patch depends on the cell length, low diffusion of active ROP, a gradient in auxin concentration, and ROP levels. Our theory provides a unique explanation linking the molecular biology to the root hair phenotypes of multiple mutants and transgenic lines, including OX-ROP, CA-rop, aux1, axr3, tip1, eto1, etr1, and the triple mutant aux1 ein2 gnom(eb). CONCLUSIONS/SIGNIFICANCE: We show how interactions between Rho GTPases (in this case ROPs) and regulatory molecules (in this case auxin) could produce characteristic subcellular patterning that subsequently affects cell shape. This has important implications for research on the morphogenesis of plants and other eukaryotes. Our results also illustrate how gradient-regulated Turing systems provide a particularly robust and flexible mechanism for pattern formation. PMID- 20016782 TI - Split-CreERT2: temporal control of DNA recombination mediated by split-Cre protein fragment complementation. AB - BACKGROUND: DNA recombination technologies such as the Cre/LoxP system advance modern biological research by allowing conditional gene regulation in vivo. However, the precise targeting of a particular cell type at a given time point has remained challenging since spatial specificity has so far depended exclusively on the promoter driving Cre recombinase expression. We have recently established split-Cre that allows DNA recombination to be controlled by coincidental activity of two promoters, thereby increasing spatial specificity of Cre-mediated DNA recombination. To allow temporal control of split-Cre-mediated DNA recombination we have now extended split-Cre by fusing split-Cre proteins with the tamoxifen inducible ERT2 domain derived from CreERT2. METHODOLOGY/PRINCIPAL FINDINGS: In the split-CreERT2 system, Cre-mediated DNA recombination is controlled by two expression cassettes as well as the time of tamoxifen application. By using two independent Cre-dependent reporters in cultured cells, the combination of NCre-ERT2+ERT2-CCre was identified as having the most favorable properties of all constructs tested, showing an induction ratio of about 10 and EC(50)-values for 4-hydroxy-tamoxifen of 10 nM to 70 nM. CONCLUSIONS/SIGNIFICANCE: These characteristics of split-CreERT2 in vitro indicate that split-CreERT2 will be well suited for inducing DNA recombination in living mice harboring LoxP-flanked alleles. In this way, split-CreERT2 will provide a new tool of modern genetics allowing spatial and temporal precise genetic access to cell populations defined by the simultaneous activity of two promoters. PMID- 20016783 TI - TGF-beta and IL-10 production by HIV-specific CD8+ T cells is regulated by CTLA-4 signaling on CD4+ T cells. AB - Immune dysregulation in HIV-1 infection is associated with increased expression of inhibitory molecules such as CTLA-4, TGF-beta, and IL-10. In this study we examined one potential mechanism for regulating TGF-beta and IL-10 expression by HIV-specific suppressor CD8+ T cells. No overlap between TGF-beta, IL-10, and IFN gamma cytokine production by HIV-specific CD8+ T cells was observed. TGF-beta positive and IL-10 positive cells were FOXP3 negative, CD25 negative, and displayed a heterogeneous surface expression of CD127. TGF-beta and IL-10 positive CD8+ T cells did not express CTLA-4. Nevertheless, CTLA-4 blockade resulted in a significant decrease in HIV-specific TGF-beta positive and IL-10 positive CD8+ T cell responses, and a concomitant increase in HIV-specific IFN gamma positive CD8+ T cell responses. Depletion of CD4+ T cells abrogated the impact of CTLA-4 on HIV-specific TGF-beta positive and IL-10 positive CD8+ T cells. Our study suggests that CTLA-4 Signaling on CD4+ T cells regulates the inhibitory functions of the HIV-specific suppressor CD8+ T cells. PMID- 20016784 TI - N-terminal deletion of peptide:N-glycanase results in enhanced deglycosylation activity. AB - Peptide:N-glycanase catalyzes the detachment of N-linked glycan chains from glycopeptides or glycoproteins by hydrolyzing the beta-aspartylglucosaminyl bond. Peptide:N-glycanase in yeast binds to Rad23p through its N-terminus. In this study, the complex formed between Peptide:N-glycanase and Rad23p was found to exhibit enhanced deglycosylation activity, which suggests an important role for this enzyme in the misfolded glycoprotein degradation pathway in vivo. To investigate the role of this enzyme in this pathway, we made stepwise deletions of the N-terminal helices of peptide:N-glycanase. Enzymatic analysis of the deletion mutants showed that deletion of the N-terminal H1 helix (Png1p-DeltaH1) enhanced the deglycosylation activity of N-glycanase towards denatured glycoproteins. In addition, this mutant exhibited high deglycosylation activity towards native glycoproteins. Dynamic simulations of the wild type and N-terminal H1 deletion mutant implied that Png1p-DeltaH1 is more flexible than wild type Png1p. The efficient deglycosylation of Png1p-DeltaH1 towards native and non native glycoproteins offers a potential biotechnological application. PMID- 20016785 TI - Genetic variance in the spinocerebellar ataxia type 2 (ATXN2) gene in children with severe early onset obesity. AB - BACKGROUND: Expansion of a CAG repeat in the coding region of exon 1 in the ATXN2 gene located in human chromosome 12q24.1 causes the neurodegenerative disease spinocerebellar ataxia type 2 (SCA2). In contrast to other polyglutamine (polyQ) disorders, the SCA2 repeat is not highly polymorphic in central European (CEU) controls with Q22 representing 90% of alleles, and Q23 contributing between 5-7% of alleles. Recently, the ATXN2 CAG repeat has been identified as a target of adaptive selection in the CEU population. Mouse lines deficient for atxn2 develop marked hyperphagia and obesity raising the possibility that loss-of-function mutations in the ATXN2 gene may be related to energy balance in humans. Some linkage studies of obesity related phenotypes such as antipsychotic induced weight gain have reported significant lod scores on chromosome 12q24. We tested the hypothesis that rare loss-of-function ATXN2 variants cause obesity analogous to rare mutations in the leptin, leptin receptor and MC4R genes. METHODOLOGY/PRINCIPAL FINDINGS: We sequenced the coding region of ATXN2 including intron-exon boundaries in 92 severely obese children with a body mass index (BMI) >3.2 standard deviations above age- and gender-adjusted means. We confirmed five previously identified single nucleotide polymorphisms (SNPs) and three new SNPs resulting in two synonymous substitutions and one intronic polymorphism. Alleles encoding >Q22 were overrepresented in our sample of obese children and contributed 15% of alleles in children identified by their parents as white. SNP rs695872 closely flanking the CAG repeat showed a greatly increased frequency of C/C homozygotes and G/C heterozygotes compared with reported frequencies in the CEU population. CONCLUSIONS/SIGNIFICANCE: Although we did not identify variants leading to novel amino acid substitutions, nonsense or frameshift mutations, this study warrants further examination of variation in the ATXN2 gene in obesity and related phenotypes in a larger case-control study with emphasis on rs695872 and CAG repeat structure. PMID- 20016786 TI - Phosphorylation-mediated control of histone chaperone ASF1 levels by Tousled-like kinases. AB - Histone chaperones are at the hub of a diverse interaction networks integrating a plethora of chromatin modifying activities. Histone H3/H4 chaperone ASF1 is a target for cell-cycle regulated Tousled-like kinases (TLKs) and both proteins cooperate during chromatin replication. However, the precise role of post translational modification of ASF1 remained unclear. Here, we identify the TLK phosphorylation sites for both Drosophila and human ASF1 proteins. Loss of TLK mediated phosphorylation triggers hASF1a and dASF1 degradation by proteasome dependent and independent mechanisms respectively. Consistent with this notion, introduction of phosphorylation-mimicking mutants inhibits hASF1a and dASF1 degradation. Human hASF1b is also targeted for proteasome-dependent degradation, but its stability is not affected by phosphorylation indicating that other mechanisms are likely to be involved in control of hASF1b levels. Together, these results suggest that ASF1 cellular levels are tightly controlled by distinct pathways and provide a molecular mechanism for post-translational regulation of dASF1 and hASF1a by TLK kinases. PMID- 20016787 TI - Paraganglioma: carotid body tumor. PMID- 20016788 TI - High grade transformation in adenoid cystic carcinoma of the parotid: report of a case with cytologic, histologic and immunohistochemical study. AB - Adenoid cystic carcinoma (ACC) constitutes about 4% of salivary epithelial tumors and is the second common malignant epithelial salivary gland tumor involving both the major and minor salivary glands. High grade transformation in ACC is a recently recognized entity with only a few cases reported in literature. We report the first case of ACC with high grade transformation involving the parotid. A 54-year-old man with a history of right parotid painful swelling from 1.5 years presented with recent increase in size of the swelling and facial paresis of 2 months duration. The locally invasive salivary neoplasm was composed predominantly of an undifferentiated carcinoma with foci of conventional ACC occupying less than 10% of tumor area. Immunohistochemical study of the undifferentiated component as compared to conventional ACC showed greater over expression of p53 and Ki-67. Her-2/Neu was negative in both components. Recognition of occasional clusters of basaloid cells and hyaline globules in association with larger poorly differentiated malignant cell population in aspiration smears can help in cytological diagnosis. The acquisition of high proliferation index and over-expression of p53 may be the probable factors involved in the pathogenesis of high grade transformation in a conventional ACC. PMID- 20016795 TI - Changes in Family Size Intentions Across Young Adulthood: A Life-Course Perspective. AB - Recent years have witnessed a resurgence in the interest in family size intentions and ideals in developed societies, partially stemming from the idea that realized fertility in these societies is lower than intended fertility. This paper addresses the question of the stability of family size intentions. Based on Heckhausen's life-span theory of control, it is hypothesized that young adults' family size intentions are likely to change as a result of their experiences in the family and occupational life domains. To study this issue, data are used from a Dutch panel survey in which respondents are questioned on their family size intentions six times over the course of 18 years. The results show that family size intentions are not stable, but are adjusted as people age. On average, the adjustment is downward, but some people do not adjust their intentions or even adjust them upwards. Much of this difference in age patterns can be explained by changes in the partner, educational, and occupational careers of young adults. Not finding a suitable partner and pursuing a career-for women-are important factors. But also the timing of the fertility career itself is of major importance. If respondents postpone having children until their thirties, they are much more likely to adjust their intentions downwards than if they start their childbearing career earlier. PMID- 20016796 TI - Reducing adverse self-medication behaviors in older adults with the Next Generation Personal Education Program (PEP-NG): Design and methodology. AB - A randomized controlled efficacy trial targeting older adults with hypertension is providing a tailored education intervention with a Next Generation Personal Education Program (PEP-NG) in primary care practices in New England. Ten participating advanced practice registered nurses (APRNs) completed online knowledge and self-efficacy measures pre-onsite training and twice more after completing a continuing education program. Patient participants self-refer in response to study recruitment brochures and posters. Twenty-four participants from each APRN practice (total N = 240) are randomly assigned by the PEP-NG software to either control (data collection and four routine APRN visits) or tailored intervention (PEP-NG interface and four focused APRN visits) conditions. Patients access the PEP-NG interface via wireless tablet and use a stylus to answer demographic, knowledge, and self-efficacy questions as well as prescription and over-the-counter self-medication practice questions. The PEP-NG analyzes patient-reported information and delivers tailored educational content. Patients' outcome measures are self-reported antihypertensive medication adherence, blood pressure, knowledge and self-efficacy concerning potential adverse self-medication practices, adverse self-medication behavior "risk" score and satisfaction with the PEP-NG and APRN provider relationship. APRN outcome measures are knowledge and self-efficacy concerning adverse self-medication practices, self-efficacy for communicating with older adults and satisfaction with the PEP-NG. Time-motion and cost-benefit analyses will be conducted. PMID- 20016797 TI - Patient preferences and satisfaction in the treatment of rheumatoid arthritis with biologic therapy. AB - Significant advances in the treatment of rheumatoid arthritis (RA) have been made over the past 10 years with the introduction of biologic therapies, such as the TNF inhibitors. With these medications, many patients with RA have seen significant improvement in symptoms, function, and quality of life. However, with the introduction of the biologics, decision-making for this chronic disease that affects up to 1% of the population has become even more complex. Patient preferences for mode and frequency of administration, and for certain risks vs benefits as well as medication beliefs are central to uptake and adherence to these medications. This review examines the current literature on patient satisfaction, adherence, and preference for biologic therapy in RA. PMID- 20016798 TI - Patient perspectives in the development and use of long-acting antipsychotics in schizophrenia: focus on olanzapine long-acting injection. AB - Schizophrenia is a chronic mental disorder generally treated with antipsychotic medication. However, non-adherence and partial adherence to antipsychotic medication treatment is common and long-acting injectable "depot" preparations of antipsychotic medications have been used as an alternative to oral medication therapy for patients for whom adherence is a clinically significant problem, as well as for the sake of convenience and in response to patient preference. Olanzapine long-acting injection (OLAI) is a new treatment option and has been approved by several regulatory agencies for the treatment of schizophrenia. OLAI is a crystalline salt formulation of olanzapine and pamoic acid. Efficacy was established in 2 double-blind randomized clinical trials of OLAI for the treatment of acute schizophrenia and for the maintenance of response. The therapeutic OLAI dosages are 150 mg q2 weeks, 210 mg q2 weeks, 300 mg q2 weeks or q4 weeks, and 405 mg q4 weeks, administered by deep intramuscular gluteal injection with a 19-gauge needle. Injection volume ranges from 1 to 2.7 mL. OLAI has essentially the same general tolerability as that of oral olanzapine; however with the depot there is the additional risk of a post-injection delirium sedation syndrome occurring at a rate of 0.07% of injections, requiring a risk management plan that includes observing the patient for 3 hours post injection. PMID- 20016799 TI - Genetic Variation of PPARs. PMID- 20016800 TI - Skeletal muscle sorbitol levels in diabetic rats with and without insulin therapy and endurance exercise training. AB - Sorbitol accumulation is postulated to play a role in skeletal muscle dysfunction associated with diabetes. The purpose of this study was to determine the effects of insulin and of endurance exercise on skeletal muscle sorbitol levels in streptozotocin-induced diabetic rats. Rats were assigned to one experimental group (control sedentary, control exercise, diabetic sedentary, diabetic exercise, diabetic sedentary no-insulin). Diabetic rats received daily subcutaneous insulin. The exercise-trained rats ran on a treadmill (1 hour, 5X/wk, for 12 weeks). Skeletal muscle sorbitol levels were the highest in the diabetic sedentary no-insulin group. Diabetic sedentary rats receiving insulin had similar sorbitol levels to control sedentary rats. Endurance exercise did not significantly affect sorbitol levels. These results indicate that insulin treatment lowers sorbitol in skeletal muscle; therefore sorbitol accumulation is probably not related to muscle dysfunction in insulin-treated diabetic individuals. Endurance exercise did not influence intramuscular sorbitol values as strongly as insulin. PMID- 20016801 TI - Procalcitonin as a Biomarker for a Bacterial Infection on Hospital Admission: A Critical Appraisal in a Cohort of Travellers with Fever after a Stay in (Sub)tropics. AB - Fever in a returned traveller may be the manifestation of a self-limiting, trivial infection but it can also presage an infection that can be rapidly progressive and lethal. We studied the diagnostic accuracy of procalcitonin (PCT) as a biomarker for a bacterial cause of fever in a cohort of 157 consecutive travellers with fever after a stay in the (sub)tropics. Elevated procalcitonin levels were observed not only in about 50% of travellers with proven bacterial infection, but also in a significant proportion of travellers with a likely infection. Using a cutoff point of 0.5 ng/mL, procalcitonin had a sensitivity of 0.52 and a specificity of 0.76 for a bacterial cause of fever on admission. Interestingly, only 1 out of 16 patients with a proven viral infection had a marginally elevated PCT concentration on admission, suggesting that an increased PCT level likely excludes a viral infection as the cause of fever. However, the diagnostic accuracy of this semiquantitative procalcitonin test for a bacterial cause of fever on admission is too poor to advocate its use in the initial clinical evaluation of fever in a setting of ill-returned travellers. PMID- 20016802 TI - Insights into the Role of GILT in HLA Class II Antigen Processing and Presentation by Melanoma. AB - Metastatic melanoma is one of the deadliest of skin cancers and is increasing in incidence. Since current treatment regimens are ineffective at controlling and/or curing the disease, novel approaches, such as immunotherapy, for treating this malignant disease are being explored. In this review, we discuss potential melanoma antigens (Ags) and their role in utilizing the HLA class II pathway to elicit tumor Ag-specific CD4+ T cell responses in order to effectively induce long-lasting CD8+ antitumor memory. We also discuss the role of endolysosomal cathepsins and Gamma-Interferon-inducible Lysosomal Thiol reductase (GILT) in Ag processing and presentation, and at enhancing CD4+ T cell recognition of melanoma cells. This review also summarizes our current knowledge on GILT and highlights a novel mechanism of GILT-mediated immune responses against melanoma cells. At the end, we propose a strategy employing GILT in the development of a potential whole cell vaccine for combating metastatic melanoma. PMID- 20016803 TI - Peroxisome proliferator-activated receptor gamma polymorphisms and coronary heart disease. AB - Single nucleotide polymorphisms (SNPs) in the peroxisome proliferator-activated receptor gamma (PPARG) gene have been associated with cardiovascular risk factors, particularly obesity and diabetes. We assessed the relationship between 4 PPARG SNPs (C-681G, C-689T, Pro12Ala, and C1431T) and coronary heart disease (CHD) in the PRIME (249 cases/494 controls, only men) and ADVANCE (1,076 cases/805 controls, men or women) studies. In PRIME, homozygote individuals for the minor allele of the PPARG C-689T, Pro12Ala, and C1431T SNPs tended to have a higher risk of CHD than homozygote individuals for the frequent allele (adjusted OR [95% CI] = 3.43 [0.96-12.27], P = .058, 3.41 [0.95-12.22], P = .060 and 5.10 [0.99-26.37], P = .050, resp.). No such association could be detected in ADVANCE. Haplotype distributions were similar in cases and control in both studies. A meta analysis on the Pro12Ala SNP, based on our data and 11 other published association studies (6,898 CHD cases/11,287 controls), revealed that there was no evidence for a significant association under the dominant model (OR = 0.99 [0.92 1.07], P = .82). However, there was a borderline association under the recessive model (OR = 1.29 [0.99-1.67], P = .06) that became significant when considering men only (OR = 1.73 [1.20-2.48], P = .003). In conclusion, the PPARG Ala12Ala genotype might be associated with a higher CHD risk in men but further confirmation studies are needed. PMID- 20016804 TI - Sentinel node detection in head and neck malignancies: innovations in radioguided surgery. AB - Sentinel node mapping is becoming a routine procedure for staging of various malignancies, because it can determine lymph node status more precisely. Due to anatomical problems, localizing sentinel nodes in the head and neck region on the basis of conventional images can be difficult. New diagnostic tools can provide better visualization of sentinel nodes. In an attempt to keep up with possible scientific progress, this article reviews new and innovative tools for sentinel node localization in this specific area. The overview comprises a short introduction of the sentinel node procedure as well as indications in the head and neck region. Then the results of SPECT/CT for sentinel node detection are described. Finally, a portable gamma camera to enable intraoperative real-time imaging with improved sentinel node detection is described. PMID- 20016805 TI - Can henna prevent ulceration in diabetic feet at high risk? PMID- 20016806 TI - Regulation of Translational Efficiency by Disparate 5' UTRs of PPARgamma Splice Variants. AB - The PPAR-gamma gene encodes for at least 7 unique transcripts due to alternative splicing of five exons in the 5'-untranslated region (UTR). The translated region is encoded by exons 1-6, which are identical in all isoforms. This study investigated the role of the 5'-UTR in regulating the efficiency with which the message is translated to protein. A coupled in vitro transcription-translation assay demonstrated that PPAR-gamma1, -gamma2, and -gamma5 are efficiently translated, whereas PPAR-gamma4 and -gamma7 are poorly translated. An in vivo reporter gene assay using each 5'-UTR upstream of the firefly luciferase gene showed that the 5'-UTRs for PPAR-gamma1, -gamma2, and -gamma4 enhanced translation, whereas the 5'-UTRs for PPAR-gamma5 and -gamma7 inhibited translation. Models of RNA secondary structure, obtained by the mfold software, were used to explain the mechanism of regulation by each 5'-UTR. In general, it was found that the translational efficiency was inversely correlated with the stability of the mRNA secondary structure, the presence of base-pairing in the consensus Kozak sequence, the number of start codons in the 5'-UTR, and the length of the 5'-UTR. A better understanding of posttranscriptional regulation of translation will allow modulation of protein levels without altering transcription. PMID- 20016807 TI - Vascular Endothelial Growth Factor plus Epidermal Growth Factor Receptor Dual Targeted Therapy in Metastatic Colorectal Cancer: Synergy or Antagonism? AB - There has been an intensive effort to develop novel therapies for the treatment of metastatic colorectal cancer (mCRC). The anti-epidermal growth factor receptor (EGFR) antibodies panitumumab and cetuximab and the anti-vascular endothelial growth factor (VEGF) antibody bevacizumab have demonstrated clinical efficacy and acceptable toxicity in the treatment of mCRC as single agents or in combination with chemotherapy. Recent clinical trials have explored the efficacy and safety of treatment regimens incorporating chemotherapy in combination with bevacizumab and either panitumumab or cetuximab in patients with mCRC. Results from the BOND 2 trial, which investigated cetuximab, bevacizumab, and chemotherapy in mCRC, provided support for this therapeutic approach. Two large randomized phase 3 trials were initiated to evaluate firstline treatment of mCRC. The Panitumumab Advanced Colorectal Cancer Evaluation (PACCE) study investigated the efficacy and safety of oxaliplatin- or irinotecan-based chemotherapy and bevacizumab with or without panitumumab; CAIRO2 assessed the efficacy and safety of capecitabine/oxaliplatin and bevacizumab with or without cetuximab. In both trials, the combination of bevacizumab, an EGFR-specific antibody, and chemotherapy in first-line treatment of mCRC was associated with increased toxicity and no improvement in patient outcome. These results suggest that these specific combinations should not be used in first-line mCRC outside investigational studies. PMID- 20016808 TI - Impact of rescue-thrombolysis during cardiopulmonary resuscitation in patients with pulmonary embolism. AB - BACKGROUND: Cardiac arrest in patients with pulmonary embolism (PE) is associated with high morbidity and mortality. Thrombolysis is expected to improve the outcome in these patients. However studies evaluating rescue-thrombolysis in patients with PE are missing, mainly due to the difficulties of clinical diagnosis of PE. We aimed to determine the success influencing factors of thrombolysis during resuscitation in patients with PE. METHODOLOGY/PRINCIPAL FINDINGS: We analyzed retrospectively the outcome of 104 consecutive patients with confirmed (n = 63) or highly suspected (n = 41) PE and monitored cardiac arrest. In all patients rtPA was administrated for thrombolysis during cardiopulmonary resuscitation. In 40 of the 104 patients (38.5%) a return of spontaneous circulation (ROSC) could be achieved successfully. Patients with ROSC received thrombolysis significantly earlier after CPR onset compared to patients without ROSC (13.6+/-1.2 min versus 24.6+/-0.8 min; p<0.001). 19 patients (47.5%) out of the 40 patients with initially successful resuscitation survived to hospital discharge. In patients with hospital discharge thrombolysis therapy was begun with a significantly shorter delay after cardiac arrest compared to all other patients (11.0+/-1.3 vs. 22.5+/-0.9 min; p<0.001). CONCLUSION: Rescue thrombolysis should be considered and started in patients with PE and cardiac arrest, as soon as possible after cardiac arrest onset. PMID- 20016809 TI - Novel Nucleoside Analogues with Fluorophores Replacing the DNA Base. AB - We describe the preparation and fluorescence properties of a set of new nucleosides in which a known hydrocarbon or oligothiophene fluorophore replaces the DNA base at C(1) of the deoxyribose moiety (see 3a - f). These compounds are potentially useful as probes in the study of the structure and dynamics of nucleic acids and their complexes with proteins. In addition, they may find use as fluorescent labels for nucleic-acid-based biomedical diagnostics methods. The fluorophores conjugated to deoxyribose at C(1) in the alpha-d-form include terphenyl, stilbene, terthiophene, benzoterthiophene, and pyrene. Also included is a non-fluorescent spacer in which cyclohexene replaces the DNA base. The nucleosides are derived from brominated fluorophore precursors and Hoffer's 2 deoxy-3,5-di-O-(p-toluoyl)-d-ribofuranosyl chloride. The emission maxima of the free nucleosides range from 345 to 536 nm. Also described are the 5' (dimethoxytrityl) 3'-O-phosphoramidite derivatives 5a - f, suitable for incorporation into oligonucleotides by automated synthesizers. PMID- 20016810 TI - Application of an Impedimetric Technique for the Detection of Lytic Infection of Salmonella spp. by Specific Phages. AB - This study was performed to evaluate the adaption of the impedimetric method to detect the lytic infection by Salmonella-specific bacteriophages and to provide a higher selectivity to this rapid method in detecting Salmonella spp. by using specific agents. Three bacteriophages and twelve strains of Salmonella spp. were tested. Each of the twelve strains was used separately to inoculate TSB together with each one of the phages. The inoculum concentration was between 10(6) and 10(7) cfu/mL, at a cell: phage ratio of 1 : 100. From the sample analysis, based on conductance (G) measurements (37 degrees C), the infection could be detected, by observation of both detection-time delay and distinct curve trends. The main conclusions were that kinetic detection by impedance microbiology with phage typing constitutes a method of determining whether a test microorganism is sensitive to the bacteriophage and a method to evaluate whether a lytic bacteriophage is present in a sample, by affecting bacterial growth rate/metabolic change. PMID- 20016811 TI - Antifungal activity of selected indigenous pseudomonas and bacillus from the soybean rhizosphere. AB - The purpose of this study was to isolate and select indigenous soil Pseudomonas and Bacillus bacteria capable of developing multiple mechanisms of action related to the biocontrol of phytopathogenic fungi affecting soybean crops. The screening procedure consisted of antagonism tests against a panel of phytopathogenic fungi, taxonomic identification, detection by PCR of several genes related to antifungal activity, in vitro detection of the antifungal products, and root colonization assays. Two isolates, identified and designated as Pseudomonas fluorescens BNM296 and Bacillus amyloliquefaciens BNM340, were selected for further studies. These isolates protected plants against the damping-off caused by Pythium ultimum and were able to increase the seedling emergence rate after inoculation of soybean seeds with each bacterium. Also, the shoot nitrogen content was higher in plants when seeds were inoculated with BNM296. The polyphasic approach of this work allowed us to select two indigenous bacterial strains that promoted the early development of soybean plants. PMID- 20016812 TI - Study of the Effect of Methyl Jasmonate Concentration on Aflatoxin B(1) Biosynthesis by Aspergillus parasiticus in Yeast Extract Sucrose Medium. AB - Aflatoxin B(1) (AFB(1)) is a carcinogenic metabolite produced by certain Aspergillus species on agricultural commodities. AFB(1) biosynthesis is affected by jasmonic acid and also by its methylester (MeJA), a plant growth regulator derived from linoleic acid. This study reports the effect of MeJA on the growth of A. parasiticus and AFB(1) output in yeast extract sucrose (YES) medium when added at three different concentrations; namely, 10(-2) M, 10(-4) M, and 10(-6) M. AFB(1) determination was performed by immunoaffinity and HPLC. MeJA at 10(-4) and 10(-6) M concentrations had no significant effect on mycelial growth but did affect AFB(1) production after the 7th day of incubation; on the 12th day, AFB(1) production was increased by 212.7% and 141.6% compared to the control samples (addition of 10(-6) M and 10(-4) M MeJA, resp.). Treatment of A. parasiticus cultures with 10(-2) M MeJA inhibited mycelial growth and AFB(1) production as well. These results suggest that the effect of MeJA on AFB(1) biosynthesis by A. parasiticus depends on the MeJA concentration used. PMID- 20016813 TI - Neural stem cells as a novel platform for tumor-specific delivery of therapeutic antibodies. AB - BACKGROUND: Recombinant monoclonal antibodies have emerged as important tools for cancer therapy. Despite the promise shown by antibody-based therapies, the large molecular size of antibodies limits their ability to efficiently penetrate solid tumors and precludes efficient crossing of the blood-brain-barrier into the central nervous system (CNS). Consequently, poorly vascularized solid tumors and CNS metastases cannot be effectively treated by intravenously-injected antibodies. The inherent tumor-tropic properties of human neural stem cells (NSCs) can potentially be harnessed to overcome these obstacles and significantly improve cancer immunotherapy. Intravenously-delivered NSCs preferentially migrate to primary and metastatic tumor sites within and outside the CNS. Therefore, we hypothesized that NSCs could serve as an ideal cellular delivery platform for targeting antibodies to malignant tumors. METHODS AND FINDINGS: As proof-of concept, we selected Herceptin (trastuzumab), a monoclonal antibody widely used to treat HER2-overexpressing breast cancer. HER2 overexpression in breast cancer is highly correlated with CNS metastases, which are inaccessible to trastuzumab therapy. Therefore, NSC-mediated delivery of trastuzumab may improve its therapeutic efficacy. Here we report, for the first time, that human NSCs can be genetically modified to secrete anti-HER2 immunoglobulin molecules. These NSC secreted antibodies assemble properly, possess tumor cell-binding affinity and specificity, and can effectively inhibit the proliferation of HER2-overexpressing breast cancer cells in vitro. We also demonstrate that immunoglobulin-secreting NSCs exhibit preferential tropism to tumor cells in vivo, and can deliver antibodies to human breast cancer xenografts in mice. CONCLUSIONS: Taken together, these results suggest that NSCs modified to secrete HER2-targeting antibodies constitute a promising novel platform for targeted cancer immunotherapy. Specifically, this NSC-mediated antibody delivery system has the potential to significantly improve clinical outcome for patients with HER2 overexpressing breast cancer. PMID- 20016814 TI - Refuge or reservoir? The potential impacts of the biofuel crop Miscanthus x giganteus on a major pest of maize. AB - BACKGROUND: Interest in the cultivation of biomass crops like the C4 grass Miscanthus x giganteus (Miscanthus) is increasing as global demand for biofuel grows. In the US, Miscanthus is promoted as a crop well-suited to the Corn Belt where it could be cultivated on marginal land interposed with maize and soybean. Interactions (direct and indirect) of Miscanthus, maize, and the major Corn Belt pest of maize, the western corn rootworm, (Diabrotica virgifera virgifera LeConte, WCR) are unknown. Adding a perennial grass/biomass crop to this system is concerning since WCR is adapted to the continuous availability of its grass host, maize (Zea mays). METHODOLOGY/PRINCIPAL FINDINGS: In a greenhouse and field study, we investigated WCR development and oviposition on Miscanthus. The suitability of Miscanthus for WCR development varied across different WCR populations. Data trends indicate that WCR populations that express behavioural resistance to crop rotation performed as well on Miscanthus as on maize. Over the entire study, total adult WCR emergence from Miscanthus (212 WCR) was 29.6% of that from maize (717 WCR). Adult dry weight was 75-80% that of WCR from maize; female emergence patterns on Miscanthus were similar to females developing on maize. There was no difference in the mean no. of WCR eggs laid at the base of Miscanthus and maize in the field. CONCLUSIONS/SIGNIFICANCE: Field oviposition and significant WCR emergence from Miscanthus raises many questions about the nature of likely interactions between Miscanthus, maize and WCR and the potential for Miscanthus to act as a refuge or reservoir for Corn Belt WCR. Responsible consideration of the benefits and risks associated with Corn Belt Miscanthus are critical to protecting an agroecosystem that we depend on for food, feed, and increasingly, fuel. Implications for European agroecosystems in which Miscanthus is being proposed are also discussed in light of the WCR's recent invasion into Europe. PMID- 20016815 TI - Voltage-dependent modulation of cardiac ryanodine receptors (RyR2) by protamine. AB - It has been reported that protamine (>10 microg/ml) blocks single skeletal RyR1 channels and inhibits RyR1-mediated Ca2+ release from sarcoplasmic reticulum microsomes. We extended these studies to cardiac RyR2 reconstituted into planar lipid bilayers. We found that protamine (0.02-20 microg/ml) added to the cytosolic surface of fully activated RyR2 affected channel activity in a voltage dependent manner. At membrane voltage (V(m); SR lumen-cytosol) = 0 mV, protamine induced conductance transitions to several intermediate states (substates) as well as full block of RyR2. At V(m)>10 mV, the substate with the highest level of conductance was predominant. Increasing V(m) from 0 to +80 mV, decreased the number of transitions and residence of the channel in this substate. The drop in current amplitude (full opening to substate) had the same magnitude at 0 and +80 mV despite the approximately 3-fold increase in amplitude of the full opening. This is more similar to rectification of channel conductance induced by other polycations than to the action of selective conductance modifiers (ryanoids, imperatoxin). A distinctive effect of protamine (which might be shared with polylysines and histones but not with non-peptidic polycations) is the activation of RyR2 in the presence of nanomolar cytosolic Ca2+ and millimolar Mg2+ levels. Our results suggest that RyRs would be subject to dual modulation (activation and block) by polycationic domains of neighboring proteins via electrostatic interactions. Understanding these interactions could be important as such anomalies may be associated with the increased RyR2-mediated Ca2+ leak observed in cardiac diseases. PMID- 20016816 TI - Secreted Gaussia luciferase as a biomarker for monitoring tumor progression and treatment response of systemic metastases. AB - BACKGROUND: Currently, only few techniques are available for quantifying systemic metastases in preclinical model. Thus techniques that can sensitively detect metastatic colonization and assess treatment response in real-time are urgently needed. To this end, we engineered tumor cells to express a naturally secreted Gaussia luciferase (Gluc), and investigated its use as a circulating biomarker for monitoring viable metastatic or primary tumor growth and their treatment responses. METHODOLOGY/PRINCIPAL FINDINGS: We first developed orthotopic primary and metastatic breast tumors with derivative of MDA-MB-231 cells expressing Gluc. We then correlated tumor burden with Gluc activity in the blood and urine along with bioluminescent imaging (BLI). Second, we utilized blood Gluc assay to monitor treatment response to lapatinib in an experimental model of systemic metastasis. We observed good correlation between the primary tumor volume and Gluc concentration in blood (R(2) = 0.84) and urine (R(2) = 0.55) in the breast tumor model. The correlation deviated as a primary tumor grew due to a reduction in viable tumor fraction. This was also supported by our mathematical models for tumor growth to compare the total and viable tumor burden in our model. In the experimental metastasis model, we found numerous brain metastases as well as systemic metastases including bone and lungs. Importantly, blood Gluc assay revealed early growth of metastatic tumors before BLI could visualize their presence. Using secreted Gluc, we localized systemic metastases by BLI and quantitatively monitored the total viable metastatic tumor burden by blood Gluc assay during the course of treatment with lapatinib, a dual tyrosine kinase inhibitor of EGFR and HER2. CONCLUSION/SIGNIFICANCE: We demonstrated secreted Gluc assay accurately reflects the amount of viable cancer cells in primary and metastatic tumors. Blood Gluc activity not only tracks metastatic tumor progression but also serves as a longitudinal biomarker for tumor response to treatments. PMID- 20016817 TI - An engineered yeast efficiently secreting penicillin. AB - This study aimed at developing an alternative host for the production of penicillin (PEN). As yet, the industrial production of this beta-lactam antibiotic is confined to the filamentous fungus Penicillium chrysogenum. As such, the yeast Hansenula polymorpha, a recognized producer of pharmaceuticals, represents an attractive alternative. Introduction of the P. chrysogenum gene encoding the non-ribosomal peptide synthetase (NRPS) delta-(L-alpha-aminoadipyl) L-cysteinyl-D-valine synthetase (ACVS) in H. polymorpha, resulted in the production of active ACVS enzyme, when co-expressed with the Bacillus subtilis sfp gene encoding a phosphopantetheinyl transferase that activated ACVS. This represents the first example of the functional expression of a non-ribosomal peptide synthetase in yeast. Co-expression with the P. chrysogenum genes encoding the cytosolic enzyme isopenicillin N synthase as well as the two peroxisomal enzymes isopenicillin N acyl transferase (IAT) and phenylacetyl CoA ligase (PCL) resulted in production of biologically active PEN, which was efficiently secreted. The amount of secreted PEN was similar to that produced by the original P. chrysogenum NRRL1951 strain (approx. 1 mg/L). PEN production was decreased over two-fold in a yeast strain lacking peroxisomes, indicating that the peroxisomal localization of IAT and PCL is important for efficient PEN production. The breakthroughs of this work enable exploration of new yeast-based cell factories for the production of (novel) beta-lactam antibiotics as well as other natural and semi-synthetic peptides (e.g. immunosuppressive and cytostatic agents), whose production involves NRPS's. PMID- 20016818 TI - Eukaryotic initiation factor 2B (eIF2B) GEF activity as a diagnostic tool for EIF2B-related disorders. AB - BACKGROUND: In recent years, the phenotypes of leukodystrophies linked to mutations in the eukaryotic initiation factor 2B genes have been extended, classically called CACH/VWM (Childhood ataxia with cntral hypomyelination/vanishing white matter disorder). The large clinical spectrum observed from the more severe antenatal forms responsible for fetal death to milder adult forms with an onset after 16 years old and restricted to slow cognitive impairment have lead to the concept of eIF2B-related disorders. The typical MRI pattern with a diffuse CSF-like aspect of the cerebral white matter can lack particularly in the adult forms whereas an increasing number of patients with clinical and MRI criteria for CACH/VWM disease but without eIF2B mutations are found. Then we propose the use of biochemical markers to help in this difficult diagnosis. The biochemical diagnosis of eIF2B-related disorder is difficult as no marker, except the recently described asialotransferrin/transferrin ratio measured in cerebrospinal fluid, has been proposed and validated until now. Decreased eIF2B GEF activity has been previously reported in lymphoblastoid cell lines from 30 eIF2B-mutated patients. Our objective was to evaluate further the utility of this marker and to validate eIF2B GEF activity in a larger cohort as a specific diagnostic test for eIF2B related disorders. METHODOLOGY/PRINCIPAL FINDINGS: We performed eIF2B GEF activity assays in cells from 63 patients presenting with different clinical forms and eIF2B mutations in comparison to controls but also to patients with defined leukodystrophies or CACH/VWM-like diseases without eIF2B mutations. We found a significant decrease of GEF activity in cells from eIF2B-mutated patients with 100% specificity and 89% sensitivity when the activity threshold was set at < or =77.5%. CONCLUSION: These results validate the measurement of eIF2B GEF activity in patients' transformed-lymphocytes as an important tool for the diagnosis of eIF2B-related disorders. PMID- 20016819 TI - Molecular exploration of the first-century Tomb of the Shroud in Akeldama, Jerusalem. AB - The Tomb of the Shroud is a first-century C.E. tomb discovered in Akeldama, Jerusalem, Israel that had been illegally entered and looted. The investigation of this tomb by an interdisciplinary team of researchers began in 2000. More than twenty stone ossuaries for collecting human bones were found, along with textiles from a burial shroud, hair and skeletal remains. The research presented here focuses on genetic analysis of the bioarchaeological remains from the tomb using mitochondrial DNA to examine familial relationships of the individuals within the tomb and molecular screening for the presence of disease. There are three mitochondrial haplotypes shared between a number of the remains analyzed suggesting a possible family tomb. There were two pathogens genetically detected within the collection of osteological samples, these were Mycobacterium tuberculosis and Mycobacterium leprae. The Tomb of the Shroud is one of very few examples of a preserved shrouded human burial and the only example of a plaster sealed loculus with remains genetically confirmed to have belonged to a shrouded male individual that suffered from tuberculosis and leprosy dating to the first century C.E. This is the earliest case of leprosy with a confirmed date in which M. leprae DNA was detected. PMID- 20016820 TI - Quantifying the extent of North American mammal extinction relative to the pre anthropogenic baseline. AB - Earth has experienced five major extinction events in the past 450 million years. Many scientists suggest we are now witnessing a sixth, driven by human impacts. However, it has been difficult to quantify the real extent of the current extinction episode, either for a given taxonomic group at the continental scale or for the worldwide biota, largely because comparisons of pre-anthropogenic and anthropogenic biodiversity baselines have been unavailable. Here, we compute those baselines for mammals of temperate North America, using a sampling standardized rich fossil record to reconstruct species-area relationships for a series of time slices ranging from 30 million to 500 years ago. We show that shortly after humans first arrived in North America, mammalian diversity dropped to become at least 15%-42% too low compared to the "normal" diversity baseline that had existed for millions of years. While the Holocene reduction in North American mammal diversity has long been recognized qualitatively, our results provide a quantitative measure that clarifies how significant the diversity reduction actually was. If mass extinctions are defined as loss of at least 75% of species on a global scale, our data suggest that North American mammals had already progressed one-fifth to more than halfway (depending on biogeographic province) towards that benchmark, even before industrialized society began to affect them. Data currently are not available to make similar quantitative estimates for other continents, but qualitative declines in Holocene mammal diversity are also widely recognized in South America, Eurasia, and Australia. Extending our methodology to mammals in these areas, as well as to other taxa where possible, would provide a reasonable way to assess the magnitude of global extinction, the biodiversity impact of extinctions of currently threatened species, and the efficacy of conservation efforts into the future. PMID- 20016821 TI - Antigen diversity in the parasitic bacterium Anaplasma phagocytophilum arises from selectively-represented, spatially clustered functional pseudogenes. AB - Anaplasma phagocytophilum is a tick-transmitted bacterial pathogen of humans and other animals, and is an obligate intracellular parasite. Throughout the course of infection, hosts acquire temporary resistance to granulocytic anaplasmosis as they develop immunity specific for the major antigen, major surface protein 2 (Msp2). However, the bacterium then utilizes a novel recombination mechanism shuffling functional pseudogenes sequentially into an expression cassette with conserved 5' and 3' ends, bypassing host immunity. Approximately 100 pseudogenes are present in the only fully sequenced human-origin HZ genome, representing the possibility for almost unlimited antigenic diversity. In the present study, we identified a select group of 20% of the A. phagocytophilum HZ msp2 pseudogenes that have matched preferentially to human, canine, and equine expression cassettes. Pseudogenes cluster predominantly in one spatial run limited to a single genomic island in less than 50% of the genome but phylogenetically related pseudogenes are neither necessarily located in close proximity on the genome nor share similar percent identity with expression cassettes. Pseudogenes near the expression cassette (and the origin) are more likely to be expressed than those farther away. Taken together, these findings suggest that there may be natural selection pressure to retain pseudogenes in one cluster near the putative origin of replication, even though global recombination shuffles pseudogenes around the genome, separating pseudogenes that share genetic origins as well as those with similar identities. PMID- 20016822 TI - Spatial and temporal analysis of gene expression during growth and fusion of the mouse facial prominences. AB - Orofacial malformations resulting from genetic and/or environmental causes are frequent human birth defects yet their etiology is often unclear because of insufficient information concerning the molecular, cellular and morphogenetic processes responsible for normal facial development. We have, therefore, derived a comprehensive expression dataset for mouse orofacial development, interrogating three distinct regions - the mandibular, maxillary and frontonasal prominences. To capture the dynamic changes in the transcriptome during face formation, we sampled five time points between E10.5-E12.5, spanning the developmental period from establishment of the prominences to their fusion to form the mature facial platform. Seven independent biological replicates were used for each sample ensuring robustness and quality of the dataset. Here, we provide a general overview of the dataset, characterizing aspects of gene expression changes at both the spatial and temporal level. Considerable coordinate regulation occurs across the three prominences during this period of facial growth and morphogenesis, with a switch from expression of genes involved in cell proliferation to those associated with differentiation. An accompanying shift in the expression of polycomb and trithorax genes presumably maintains appropriate patterns of gene expression in precursor or differentiated cells, respectively. Superimposed on the many coordinated changes are prominence-specific differences in the expression of genes encoding transcription factors, extracellular matrix components, and signaling molecules. Thus, the elaboration of each prominence will be driven by particular combinations of transcription factors coupled with specific cell:cell and cell:matrix interactions. The dataset also reveals several prominence-specific genes not previously associated with orofacial development, a subset of which we externally validate. Several of these latter genes are components of bidirectional transcription units that likely share cis-acting sequences with well-characterized genes. Overall, our studies provide a valuable resource for probing orofacial development and a robust dataset for bioinformatic analysis of spatial and temporal gene expression changes during embryogenesis. PMID- 20016823 TI - Cranial anatomy of the earliest marsupials and the origin of opossums. AB - BACKGROUND: The early evolution of living marsupials is poorly understood in part because the early offshoots of this group are known almost exclusively from jaws and teeth. Filling this gap is essential for a better understanding of the phylogenetic relationships among living marsupials, the biogeographic pathways that led to their current distribution as well as the successive evolutionary steps that led to their current diversity, habits and various specializations that distinguish them from placental mammals. METHODOLOGY/PRINCIPAL FINDINGS: Here we report the first skull of a 55 million year old peradectid marsupial from the early Eocene of North America and exceptionally preserved skeletons of an Oligocene herpetotheriid, both representing critical groups to understand early marsupial evolution. A comprehensive phylogenetic cladistic analysis of Marsupialia including the new findings and close relatives of marsupials show that peradectids are the sister group of living opossums and herpetotheriids are the sister group of all living marsupials. CONCLUSIONS/SIGNIFICANCE: The results imply that North America played an important role in early Cenozoic marsupial evolutionary history and may have even been the center of origin of living marsupials and opossums. New data from the herpetotheriid postcranium support the view that the ancestral morphotype of Marsupialia was more terrestrial than opossums are. The resolution of the phylogenetic position of peradectids reveals an older calibration point for molecular estimates of divergence times among living marsupials than those currently used. PMID- 20016824 TI - Kisspeptin signalling in the hypothalamic arcuate nucleus regulates GnRH pulse generator frequency in the rat. AB - BACKGROUND: Kisspeptin and its G protein-coupled receptor (GPR) 54 are essential for activation of the hypothalamo-pituitary-gonadal axis. In the rat, the kisspeptin neurons critical for gonadotropin secretion are located in the hypothalamic arcuate (ARC) and anteroventral periventricular (AVPV) nuclei. As the ARC is known to be the site of the gonadotropin-releasing hormone (GnRH) pulse generator we explored whether kisspeptin-GPR54 signalling in the ARC regulates GnRH pulses. METHODOLOGY/PRINCIPAL FINDINGS: We examined the effects of kisspeptin-10 or a selective kisspeptin antagonist administration intra-ARC or intra-medial preoptic area (mPOA), (which includes the AVPV), on pulsatile luteinizing hormone (LH) secretion in the rat. Ovariectomized rats with subcutaneous 17beta-estradiol capsules were chronically implanted with bilateral intra-ARC or intra-mPOA cannulae, or intra-cerebroventricular (icv) cannulae and intravenous catheters. Blood samples were collected every 5 min for 5-8 h for LH measurement. After 2 h of control blood sampling, kisspeptin-10 or kisspeptin antagonist was administered via pre-implanted cannulae. Intranuclear administration of kisspeptin-10 resulted in a dose-dependent increase in circulating levels of LH lasting approximately 1 h, before recovering to a normal pulsatile pattern of circulating LH. Both icv and intra-ARC administration of kisspeptin antagonist suppressed LH pulse frequency profoundly. However, intra mPOA administration of kisspeptin antagonist did not affect pulsatile LH secretion. CONCLUSIONS/SIGNIFICANCE: These data are the first to identify the arcuate nucleus as a key site for kisspeptin modulation of LH pulse frequency, supporting the notion that kisspeptin-GPR54 signalling in this region of the mediobasal hypothalamus is a critical neural component of the hypothalamic GnRH pulse generator. PMID- 20016825 TI - Testosterone administration decreases generosity in the ultimatum game. AB - How do human beings decide when to be selfish or selfless? In this study, we gave testosterone to 25 men to establish its impact on prosocial behaviors in a double blind within-subjects design. We also confirmed participants' testosterone levels before and after treatment through blood draws. Using the Ultimatum Game from behavioral economics, we find that men with artificially raised T, compared to themselves on placebo, were 27% less generous towards strangers with money they controlled (95% CI placebo: (1.70, 2.72); 95% CI T: (.98, 2.30)). This effect scales with a man's level of total-, free-, and dihydro-testosterone (DHT). Men in the lowest decile of DHT were 560% more generous than men in the highest decile of DHT. We also found that men with elevated testosterone were more likely to use their own money punish those who were ungenerous toward them. Our results continue to hold after controlling for altruism. We conclude that elevated testosterone causes men to behave antisocially. PMID- 20016826 TI - Protection of Salvia miltiorrhizae to the spleen and thymus of rats with severe acute pancreatitis or obstructive jaundice. AB - OBJECTIVE: To investigate the therapeutic effects and mechanism of Salvia miltiorrhizae in the treatment of SAP and OJ. METHODS: A total of 288 rats were used for SAP- and OJ-associated experiments. The rats were randomly divided into sham-operated group, model control group and treated group. The mortality rates of rats, contents of endotoxin and PLA(2) in blood, pathological changes of different indexes in spleen and thymus were observed. RESULTS: The contents of endotoxin and PLA2 in treated group were significantly lower than those in model control group.The pathological severity scores of spleen and thymus of SAP rats as well as that of spleen of OJ rats in treated groups were significantly lower than those in model control groups (P < .05). The staining intensity as well as the product of the staining intensity and positive rate of Bax protein of spleen in model control group were significantly higher than those in treated groups (P < .01) , and the apoptosis index of spleen in treated group was significantly lower than that in model control group (P < .01). CONCLUSION: Salvia miltiorrhizae exerts protective effects on the spleen and thymus of SAP rats and spleen of OJ rats. PMID- 20016827 TI - Applied climate-change analysis: the climate wizard tool. AB - BACKGROUND: Although the message of "global climate change" is catalyzing international action, it is local and regional changes that directly affect people and ecosystems and are of immediate concern to scientists, managers, and policy makers. A major barrier preventing informed climate-change adaptation planning is the difficulty accessing, analyzing, and interpreting climate-change information. To address this problem, we developed a powerful, yet easy to use, web-based tool called Climate Wizard (http://ClimateWizard.org) that provides non climate specialists with simple analyses and innovative graphical depictions for conveying how climate has and is projected to change within specific geographic areas throughout the world. METHODOLOGY/PRINCIPAL FINDINGS: To demonstrate the Climate Wizard, we explored historic trends and future departures (anomalies) in temperature and precipitation globally, and within specific latitudinal zones and countries. We found the greatest temperature increases during 1951-2002 occurred in northern hemisphere countries (especially during January-April), but the latitude of greatest temperature change varied throughout the year, sinusoidally ranging from approximately 50 degrees N during February-March to 10 degrees N during August-September. Precipitation decreases occurred most commonly in countries between 0-20 degrees N, and increases mostly occurred outside of this latitudinal region. Similarly, a quantile ensemble analysis based on projections from 16 General Circulation Models (GCMs) for 2070-2099 identified the median projected change within countries, which showed both latitudinal and regional patterns in projected temperature and precipitation change. CONCLUSIONS/SIGNIFICANCE: The results of these analyses are consistent with those reported by the Intergovernmental Panel on Climate Change, but at the same time, they provide examples of how Climate Wizard can be used to explore regionally- and temporally-specific analyses of climate change. Moreover, Climate Wizard is not a static product, but rather a data analysis framework designed to be used for climate change impact and adaption planning, which can be expanded to include other information, such as downscaled future projections of hydrology, soil moisture, wildfire, vegetation, marine conditions, disease, and agricultural productivity. PMID- 20016828 TI - A large scale analysis of information-theoretic network complexity measures using chemical structures. AB - This paper aims to investigate information-theoretic network complexity measures which have already been intensely used in mathematical- and medicinal chemistry including drug design. Numerous such measures have been developed so far but many of them lack a meaningful interpretation, e.g., we want to examine which kind of structural information they detect. Therefore, our main contribution is to shed light on the relatedness between some selected information measures for graphs by performing a large scale analysis using chemical networks. Starting from several sets containing real and synthetic chemical structures represented by graphs, we study the relatedness between a classical (partition-based) complexity measure called the topological information content of a graph and some others inferred by a different paradigm leading to partition-independent measures. Moreover, we evaluate the uniqueness of network complexity measures numerically. Generally, a high uniqueness is an important and desirable property when designing novel topological descriptors having the potential to be applied to large chemical databases. PMID- 20016829 TI - Culture modulates eye-movements to visual novelty. AB - BACKGROUND: When viewing complex scenes, East Asians attend more to contexts whereas Westerners attend more to objects, reflecting cultural differences in holistic and analytic visual processing styles respectively. This eye-tracking study investigated more specific mechanisms and the robustness of these cultural biases in visual processing when salient changes in the objects and backgrounds occur in complex pictures. METHODOLOGY/PRINCIPAL FINDINGS: Chinese Singaporean (East Asian) and Caucasian US (Western) participants passively viewed pictures containing selectively changing objects and background scenes that strongly captured participants' attention in a data-driven manner. We found that although participants from both groups responded to object changes in the pictures, there was still evidence for cultural divergence in eye-movements. The number of object fixations in the US participants was more affected by object change than in the Singapore participants. Additionally, despite the picture manipulations, US participants consistently maintained longer durations for both object and background fixations, with eye-movements that generally remained within the focal objects. In contrast, Singapore participants had shorter fixation durations with eye-movements that alternated more between objects and backgrounds. CONCLUSIONS/SIGNIFICANCE: The results demonstrate a robust cultural bias in visual processing even when external stimuli draw attention in an opposite manner to the cultural bias. These findings also extend previous studies by revealing more specific, but consistent, effects of culture on the different aspects of visual attention as measured by fixation duration, number of fixations, and saccades between objects and backgrounds. PMID- 20016830 TI - Chagas disease, migration and community settlement patterns in Arequipa, Peru. AB - BACKGROUND: Chagas disease is one of the most important neglected tropical diseases in the Americas. Vectorborne transmission of Chagas disease has been historically rare in urban settings. However, in marginal communities near the city of Arequipa, Peru, urban transmission cycles have become established. We examined the history of migration and settlement patterns in these communities, and their connections to Chagas disease transmission. METHODOLOGY/PRINCIPAL FINDINGS: This was a qualitative study that employed focus group discussions and in-depth interviews. Five focus groups and 50 in-depth interviews were carried out with 94 community members from three shantytowns and two traditional towns near Arequipa, Peru. Focus groups utilized participatory methodologies to explore the community's mobility patterns and the historical and current presence of triatomine vectors. In-depth interviews based on event history calendars explored participants' migration patterns and experience with Chagas disease and vectors. Focus group data were analyzed using participatory analysis methodologies, and interview data were coded and analyzed using a grounded theory approach. Entomologic data were provided by an ongoing vector control campaign. We found that migrants to shantytowns in Arequipa were unlikely to have brought triatomines to the city upon arrival. Frequent seasonal moves, however, took shantytown residents to valleys surrounding Arequipa where vectors are prevalent. In addition, the pattern of settlement of shantytowns and the practice of raising domestic animals by residents creates a favorable environment for vector proliferation and dispersal. Finally, we uncovered a phenomenon of population loss and replacement by low-income migrants in one traditional town, which created the human settlement pattern of a new shantytown within this traditional community. CONCLUSIONS/SIGNIFICANCE: The pattern of human migration is therefore an important underlying determinant of Chagas disease risk in and around Arequipa. Frequent seasonal migration by residents of peri-urban shantytowns provides a path of entry of vectors into these communities. Changing demographic dynamics of traditional towns are also leading to favorable conditions for Chagas disease transmission. Control programs must include surveillance for infestation in communities assumed to be free of vectors. PMID- 20016831 TI - Dopamine D2 receptor stimulation potentiates PolyQ-Huntingtin-induced mouse striatal neuron dysfunctions via Rho/ROCK-II activation. AB - BACKGROUND: Huntington's disease (HD) is a polyglutamine-expanded related neurodegenerative disease. Despite the ubiquitous expression of expanded, polyQ Huntingtin (ExpHtt) in the brain, striatal neurons present a higher susceptibility to the mutation. A commonly admitted hypothesis is that Dopaminergic inputs participate to this vulnerability. We previously showed that D2 receptor stimulation increased aggregate formation and neuronal death induced by ExpHtt in primary striatal neurons in culture, and chronic D2 antagonist treatment protects striatal dysfunctions induced by ExpHtt in a lentiviral induced model system in vivo. The present work was designed to elucidate the signalling pathways involved, downstream D2 receptor (D2R) stimulation, in striatal vulnerability to ExpHtt. METHODOLOGY/PRINCIPAL FINDINGS: Using primary striatal neurons in culture, transfected with a tagged-GFP version of human exon 1 ExpHtt, and siRNAs against D2R or D1R, we confirm that DA potentiates neuronal dysfunctions via D2R but not D1R stimulation. We demonstrate that D2 agonist treatment induces neuritic retraction and growth cone collapse in Htt- and ExpHtt expressing neurons. We then tested a possible involvement of the Rho/ROCK signalling pathway, which plays a key role in the dynamic of the cytoskeleton, in these processes. The pharmacological inhibitors of ROCK (Y27632 and Hydroxyfasudil), as well as siRNAs against ROCK-II, reversed D2-related effects on neuritic retraction and growth cone collapse. We show a coupling between D2 receptor stimulation and Rho activation, as well as hyperphosphorylation of Cofilin, a downstream effector of ROCK-II pathway. Importantly, D2 agonist mediated potentiation of aggregate formation and neuronal death induced by ExpHtt, was totally reversed by Y27632 and Hydroxyfasudil and ROCK-II siRNAs. CONCLUSIONS/SIGNIFICANCE: Our data provide the first demonstration that D2R induced vulnerability in HD is critically linked to the activation of the Rho/ROCK signalling pathway. The inclusion of Rho/ROCK inhibitors could be an interesting therapeutic option aimed at forestalling the onset of the disease. PMID- 20016832 TI - Release from cross-orientation suppression facilitates 3D shape perception. AB - Cross-orientation suppression (COS) in striate cortex has been implicated in the efficient encoding of visual stimuli. We show that release from COS facilitates the decoding of 3-D shape. In planar surfaces overlaid with textures, slanting the surface can increase the visibility of the component parallel to the slant. Since this component provides the orientation flows that signify 3-D shape, the enhancement of visibility facilitates 3-D slant perception. Contrast thresholds reveal that this enhancement results from a decrease in COS when 3-D slant creates a frequency mismatch between texture components. We show that coupling compressive nonlinearities in LGN neurons with expansive nonlinearities in cortical neurons can model the frequency-specific component of suppression. PMID- 20016834 TI - Efficient assembly and secretion of recombinant subviral particles of the four dengue serotypes using native prM and E proteins. AB - BACKGROUND: Flavivirus infected cells produce infectious virions and subviral particles, both of which are formed by the assembly of prM and E envelope proteins and are believed to undergo the same maturation process. Dengue recombinant subviral particles have been produced in cell cultures with either modified or chimeric proteins but not using the native forms of prM and E. METHODOLOGY/PRINCIPAL FINDINGS: We have used a codon optimization strategy to obtain an efficient expression of native viral proteins and production of recombinant subviral particles (RSPs) for all four dengue virus (DV) serotypes. A stable HeLa cell line expressing DV1 prME was established (HeLa-prME) and RSPs were analyzed by immunofluorescence and transmission electron microscopy. We found that E protein is mainly present in the endoplasmic reticulum (ER) where assembly of RSPs could be observed. Biochemical characterization of DV1 RSPs secretion revealed both prM protein cleavage and homodimerization of E proteins before their release into the supernatant, indicating that RSPs undergo a similar maturation process as dengue virus. Pulse chase experiment showed that 8 hours are required for the secretion of DV1 RSPs. We have used HeLa-prME to develop a semi-quantitative assay and screened a human siRNA library targeting genes involved in membrane trafficking. Knockdown of 23 genes resulted in a significant reduction in DV RSP secretion, whereas for 22 others we observed an increase of RSP levels in cell supernatant. CONCLUSIONS/SIGNIFICANCE: Our data describe the efficient production of RSPs containing native prM and E envelope proteins for all dengue serotypes. Dengue RSPs and corresponding producing cell lines are safe and novel tools that can be used in the study of viral egress as well as in the development of vaccine and drugs against dengue virus. PMID- 20016833 TI - Mitochondrial mislocalization underlies Abeta42-induced neuronal dysfunction in a Drosophila model of Alzheimer's disease. AB - The amyloid-beta 42 (Abeta42) is thought to play a central role in the pathogenesis of Alzheimer's disease (AD). However, the molecular mechanisms by which Abeta42 induces neuronal dysfunction and degeneration remain elusive. Mitochondrial dysfunctions are implicated in AD brains. Whether mitochondrial dysfunctions are merely a consequence of AD pathology, or are early seminal events in AD pathogenesis remains to be determined. Here, we show that Abeta42 induces mitochondrial mislocalization, which contributes to Abeta42-induced neuronal dysfunction in a transgenic Drosophila model. In the Abeta42 fly brain, mitochondria were reduced in axons and dendrites, and accumulated in the somata without severe mitochondrial damage or neurodegeneration. In contrast, organization of microtubule or global axonal transport was not significantly altered at this stage. Abeta42-induced behavioral defects were exacerbated by genetic reductions in mitochondrial transport, and were modulated by cAMP levels and PKA activity. Levels of putative PKA substrate phosphoproteins were reduced in the Abeta42 fly brains. Importantly, perturbations in mitochondrial transport in neurons were sufficient to disrupt PKA signaling and induce late-onset behavioral deficits, suggesting a mechanism whereby mitochondrial mislocalization contributes to Abeta42-induced neuronal dysfunction. These results demonstrate that mislocalization of mitochondria underlies the pathogenic effects of Abeta42 in vivo. PMID- 20016835 TI - Rhodopsin molecular evolution in mammals inhabiting low light environments. AB - The ecological radiation of mammals to inhabit a variety of light environments is largely attributed to adaptive changes in their visual systems. Visual capabilities are conferred by anatomical features of the eyes as well as the combination and properties of their constituent light sensitive pigments. To test whether evolutionary switches to different niches characterized by dim-light conditions coincided with molecular adaptation of the rod pigment rhodopsin, we sequenced the rhodopsin gene in twenty-two mammals including several bats and subterranean mole-rats. We compared these to thirty-seven published mammal rhodopsin sequences, from species with divergent visual ecologies, including nocturnal, diurnal and aquatic groups. All taxa possessed an intact functional rhodopsin; however, phylogenetic tree reconstruction recovered a gene tree in which rodents were not monophyletic, and also in which echolocating bats formed a monophyletic group. These conflicts with the species tree appear to stem from accelerated evolution in these groups, both of which inhabit low light environments. Selection tests confirmed divergent selection pressures in the clades of subterranean rodents and bats, as well as in marine mammals that live in turbid conditions. We also found evidence of divergent selection pressures among groups of bats with different sensory modalities based on vision and echolocation. Sliding window analyses suggest most changes occur in transmembrane domains, particularly obvious within the pinnipeds; however, we found no obvious pattern between photopic niche and predicted spectral sensitivity based on known critical amino acids. This study indicates that the independent evolution of rhodopsin vision in ecologically specialised groups of mammals has involved molecular evolution at the sequence level, though such changes might not mediate spectral sensitivity directly. PMID- 20016836 TI - Finding of the low molecular weight inhibitors of resuscitation promoting factor enzymatic and resuscitation activity. AB - BACKGROUND: Resuscitation promoting factors (RPF) are secreted proteins involved in reactivation of dormant actinobacteria, including Mycobacterium tuberculosis. They have been considered as prospective targets for the development of new anti tuberculosis drugs preventing reactivation of dormant tubercle bacilli, generally associated with latent tuberculosis. However, no inhibitors of Rpf activity have been reported so far. The goal of this study was to find low molecular weight compounds inhibiting the enzymatic and biological activities of Rpfs. METHODOLOGY/PRINCIPAL FINDINGS: Here we describe a novel class of 2 nitrophenylthiocyanates (NPT) compounds that inhibit muralytic activity of Rpfs with IC(50) 1-7 microg/ml. Fluorescence studies revealed interaction of active NPTs with the internal regions of the Rpf molecule. Candidate inhibitors of Rpf enzymatic activity showed a bacteriostatic effect on growth of Micrococcus luteus (in which Rpf is essential for growth protein) at concentrations close to IC(50). The candidate compounds suppressed resuscitation of dormant ("non-culturable") cells of M. smegmatis at 1 microg/ml or delayed resuscitation of dormant M. tuberculosis obtained in laboratory conditions at 10 microg/ml. However, they did not inhibit growth of active mycobacteria under these concentrations. CONCLUSIONS/SIGNIFICANCE: NPT are the first example of low molecular weight compounds that inhibit the enzymatic and biological activities of Rpf proteins. PMID- 20016837 TI - Stable patterns of gene expression regulating carbohydrate metabolism determined by geographic ancestry. AB - BACKGROUND: Individuals of African descent in the United States suffer disproportionately from diseases with a metabolic etiology (obesity, metabolic syndrome, and diabetes), and from the pathological consequences of these disorders (hypertension and cardiovascular disease). METHODOLOGY/PRINCIPAL FINDINGS: Using a combination of genetic/genomic and bioinformatics approaches, we identified a large number of genes that were both differentially expressed between American subjects self-identified to be of either African or European ancestry and that also contained single nucleotide polymorphisms that distinguish distantly related ancestral populations. Several of these genes control the metabolism of simple carbohydrates and are direct targets for the SREBP1, a metabolic transcription factor also differentially expressed between our study populations. CONCLUSIONS/SIGNIFICANCE: These data support the concept of stable patterns of gene transcription unique to a geographic ancestral lineage. Differences in expression of several carbohydrate metabolism genes suggest both genetic and transcriptional mechanisms contribute to these patterns and may play a role in exacerbating the disproportionate levels of obesity, diabetes, and cardiovascular disease observed in Americans with African ancestry. PMID- 20016838 TI - Immediate risk for cardiovascular events and suicide following a prostate cancer diagnosis: prospective cohort study. AB - BACKGROUND: Stressful life events have been shown to be associated with altered risk of various health consequences. The aim of the present study was to investigate whether the emotional stress evoked by a prostate cancer diagnosis increases the immediate risks of cardiovascular events and suicide. METHODS AND FINDINGS: We conducted a prospective cohort study by following all men in Sweden who were 30 y or older (n = 4,305,358) for a diagnosis of prostate cancer (n = 168,584) and their subsequent occurrence of cardiovascular events and suicide between January 1, 1961 and December 31, 2004. We used Poisson regression models to calculate relative risks (RRs) and 95% confidence intervals (CIs) of cardiovascular events and suicide among men who had prostate cancer diagnosed within 1 y to men without any cancer diagnosis. The risks of cardiovascular events and suicide were elevated during the first year after prostate cancer diagnosis, particularly during the first week. Before 1987, the RR of fatal cardiovascular events was 11.2 (95% CI 10.4-12.1) during the first week and 1.9 (95% CI 1.9-2.0) during the first year after diagnosis. From 1987, the RR for cardiovascular events, nonfatal and fatal combined, was 2.8 (95% CI 2.5-3.2) during the first week and 1.3 (95% CI 1.3-1.3) during the first year after diagnosis. While the RR of cardiovascular events declined, the RR of suicide was stable over the entire study period: 8.4 (95% CI 1.9-22.7) during the first week and 2.6 (95% CI 2.1-3.0) during the first year after diagnosis. Men 54 y or younger at cancer diagnosis demonstrated the highest RRs of both cardiovascular events and suicide. A limitation of the present study is the lack of tumor stage data, which precluded possibilities of investigating the potential impact of the disease severity on the relationship between a recent diagnosis of prostate cancer and the risks of cardiovascular events and suicide. In addition, we cannot exclude residual confounding as a possible explanation. CONCLUSIONS: Men newly diagnosed with prostate cancer are at increased risks for cardiovascular events and suicide. Future studies with detailed disease characteristic data are warranted. PMID- 20016839 TI - Th17: a new participant in gut dysfunction in mice infected with Trichinella spiralis. AB - Trichinella spiralis infection in rodents is a well-known model of intestinal inflammation associated with hypermotility. Our aim was to elucidate if Th17 cells were involved in the development of gastrointestinal hypermotility in this experimental model. Intestinal inflammation was observed by hematoxylin-eosin (HE) staining. Jejunal smooth muscle contractility was investigated in response to acetylcholine (Ach). The effects of IL-17 on jejunum smooth muscle contractility were explored. Flow cytometry was used to analyze the proportion of Th17 cells in jejunum. The levels of IL-17, IL-23, and TGF-beta1 in jejunum were measured by Western blot. Our results showed that the inflammation in jejunum was severe at 2 weeks postinfection (PI), which was not discernible at 8 weeks PI. Jejunal smooth muscle contractility was increased at 2 weeks PI and kept higher at 12 weeks PI. The proportion of Th17 cells and the expression of IL-17 were upregulated in jejunum at 2 weeks PI and normalized at 8 weeks PI. When jejunual smooth muscle strips were cultured with IL-17, contractions elicited by Ach were enhanced in a concentration-dependent manner. Our data suggest that Th17 cells are increased during acute infection with Trichinella spiralis and IL-17 may contribute to jejunal muscle contractility in mice. PMID- 20016840 TI - Cardiac glycosides induce cell death in human cells by inhibiting general protein synthesis. AB - BACKGROUND: Cardiac glycosides are Na(+)/K(+)-pump inhibitors widely used to treat heart failure. They are also highly cytotoxic, and studies have suggested specific anti-tumor activity leading to current clinical trials in cancer patients. However, a definitive demonstration of this putative anti-cancer activity and the underlying molecular mechanism has remained elusive. METHODOLOGY/PRINCIPAL FINDINGS: Using an unbiased transcriptomics approach, we found that cardiac glycosides inhibit general protein synthesis. Protein synthesis inhibition and cytotoxicity were not specific for cancer cells as they were observed in both primary and cancer cell lines. These effects were dependent on the Na(+)/K(+)-pump as they were rescued by expression of a cardiac glycoside resistant Na(+)/K(+)-pump. Unlike human cells, rodent cells are largely resistant to cardiac glycosides in vitro and mice were found to tolerate extremely high levels. CONCLUSIONS/SIGNIFICANCE: The physiological difference between human and mouse explains the previously observed sensitivity of human cancer cells in mouse xenograft experiments. Thus, published mouse xenograft models used to support anti-tumor activity for these drugs require reevaluation. Our finding that cardiac glycosides inhibit protein synthesis provides a mechanism for the cytotoxicity of CGs and raises concerns about ongoing clinical trials to test CGs as anti-cancer agents in humans. PMID- 20016841 TI - Possible fruit protein effects on primate communities in madagascar and the neotropics. AB - BACKGROUND: The ecological factors contributing to the evolution of tropical vertebrate communities are still poorly understood. Primate communities of the tropical Americas have fewer folivorous but more frugivorous genera than tropical regions of the Old World and especially many more frugivorous genera than Madagascar. Reasons for this phenomenon are largely unexplored. We developed the hypothesis that Neotropical fruits have higher protein concentrations than fruits from Madagascar and that the higher representation of frugivorous genera in the Neotropics is linked to high protein concentrations in fruits. Low fruit protein concentrations in Madagascar would restrict the evolution of frugivores in Malagasy communities. METHODOLOGY/PRINCIPAL FINDINGS: We reviewed the literature for nitrogen concentrations in fruits from the Neotropics and from Madagascar, and analyzed fruits from an additional six sites in the Neotropics and six sites in Madagascar. Fruits from the Neotropical sites contain significantly more nitrogen than fruits from the Madagascar sites. Nitrogen concentrations in New World fruits are above the concentrations to satisfy nitrogen requirements of primates, while they are at the lower end or below the concentrations to cover primate protein needs in Madagascar. CONCLUSIONS/SIGNIFICANCE: Fruits at most sites in the Neotropics contain enough protein to satisfy the protein needs of primates. Thus, selection pressure to develop new adaptations for foods that are difficult to digest (such as leaves) may have been lower in the Neotropics than in Madagascar. The low nitrogen concentrations in fruits from Madagascar may contribute to the almost complete absence of frugivorous primate species on this island. PMID- 20016842 TI - Extracellular tumor-related mRNA in plasma of lymphoma patients and survival implications. AB - BACKGROUND: We studied anomalous extracellular mRNAs in plasma from patients with diffuse large B-cell lymphoma (DLBCL) and their survival implications. mRNAs studied have been reported in the literature as markers of poor (BCL2, CCND2, MYC) and favorable outcome (LMO2, BCL6, FN1) in tumors. These markers were also analyzed in lymphoma tissues to test possible associations with their presence in plasma. METHODOLOGY/PRINCIPAL FINDINGS: mRNA from 42 plasma samples and 12 tumors from patients with DLBCL was analyzed by real-time PCR. Samples post-treatment were studied. The immunohistochemistry of BCL2 and BCL6 was defined. Presence of circulating tumor cells was determined by analyzing the clonality of the immunoglobulin heavy-chain genes by PCR. In DLBCL, MYC mRNA was associated with short overall survival. mRNA targets with unfavorable outcome in tumors were associated with characteristics indicative of poor prognosis, with partial treatment response and with short progression-free survival in patients with complete response. In patients with low IPI score, unfavorable mRNA targets were related to shorter overall survival, partial response, high LDH levels and death. mRNA disappeared in post-treatment samples of patients with complete response, and persisted in those with partial response or death. No associations were found between circulating tumor cells and plasma mRNA. Absence of BCL6 protein in tumors was associated with presence of unfavorable plasma mRNA. CONCLUSIONS/SIGNIFICANCE: Through a non-invasive procedure, tumor-derived mRNAs can be obtained in plasma. mRNA detected in plasma did not proceed from circulating tumor cells. In our study, unfavorable targets in plasma were associated with poor prognosis in B-cell lymphomas, mainly MYC mRNA. Moreover, the unfavorable targets in plasma could help us to classify patients with poor outcome within the good prognosis group according to IPI. PMID- 20016843 TI - Neural population-level memory traces in the mouse hippocampus. AB - One of the fundamental goals in neurosciences is to elucidate the formation and retrieval of brain's associative memory traces in real-time. Here, we describe real-time neural ensemble transient dynamics in the mouse hippocampal CA1 region and demonstrate their relationships with behavioral performances during both learning and recall. We employed the classic trace fear conditioning paradigm involving a neutral tone followed by a mild foot-shock 20 seconds later. Our large-scale recording and decoding methods revealed that conditioned tone responses and tone-shock association patterns were not present in CA1 during the first pairing, but emerged quickly after multiple pairings. These encoding patterns showed increased immediate-replay, correlating tightly with increased immediate-freezing during learning. Moreover, during contextual recall, these patterns reappeared in tandem six-to-fourteen times per minute, again correlating tightly with behavioral recall. Upon traced tone recall, while various fear memories were retrieved, the shock traces exhibited a unique recall-peak around the 20-second trace interval, further signifying the memory of time for the expected shock. Therefore, our study has revealed various real-time associative memory traces during learning and recall in CA1, and demonstrates that real-time memory traces can be decoded on a moment-to-moment basis over any single trial. PMID- 20016844 TI - The secret life of oilbirds: new insights into the movement ecology of a unique avian frugivore. AB - BACKGROUND: Steatornis caripensis (the oilbird) is a very unusual bird. It supposedly never sees daylight, roosting in huge aggregations in caves during the day and bringing back fruit to the cave at night. As a consequence a large number of the seeds from the fruit they feed upon germinate in the cave and spoil. METHODOLOGY/PRINCIPAL FINDINGS: Here we use newly developed GPS/acceleration loggers with remote UHF readout to show that several assumptions about the behaviour of Steatornis caripensis need to be revised. On average, they spend only every 3(rd) day in a cave, individuals spent most days sitting quietly in trees in the rainforest where they regurgitate seeds. CONCLUSIONS/SIGNIFICANCE: This provides new data on the extent of seed dispersal and the movement ecology of Steatornis caripensis. It suggests that Steatornis caripensis is perhaps the most important long-distance seed disperser in Neotropical forests. We also show that colony-living comes with high activity costs to individuals. PMID- 20016845 TI - Review of extended-release niacin/laropiprant fixed combination in the treatment of mixed dyslipidemia and primary hypercholesterolemia. AB - Although statins reduce cardiovascular morbidity and mortality further risk reduction is needed. In this respect low HDL-cholesterol concentrations and/or elevated triglyceride concentrations may be potential treatment targets. Niacin (nicotinic acid) is an effective drug which increases the plasma concentration of high-density lipoprotein (HDL)-cholesterol and decreases the concentration of low density lipoprotein (LDL)-cholesterol, triglycerides and lipoprotein(a). Clinical studies indicate that niacin can significantly reduce the risk for cardiovascular events. However, niacin is not very commonly used because of significant side effects (especially flushing). Laropiprant is a potent selective antagonist of PGD2-receptor subtype-1 and can thus reduce niacin-induced flushing. Although the addition of laropiprant will reduce the frequency of flushing, it will not completely eliminate this side effect. Laropiprant does not change the effect of niacin on lipids or other side effects of niacin (ie, gastro-intestinal problems, glucose elevation). The combination of niacin with laropiprant may therefore enable use of niacin at higher doses and therefore exploit the full potential of the drug. Endpoint studies that will be published over the next few years will show whether this treatment modality also translates into clinical effect in patients treated with statins. Until publication of these studies niacin/laropiprant should be used only in high-risk patients not achieving lipid goals on statins. PMID- 20016846 TI - Spatial predictions of Rhodesian Human African Trypanosomiasis (sleeping sickness) prevalence in Kaberamaido and Dokolo, two newly affected districts of Uganda. AB - The continued northwards spread of Rhodesian sleeping sickness or Human African Trypanosomiasis (HAT) within Uganda is raising concerns of overlap with the Gambian form of the disease. Disease convergence would result in compromised diagnosis and treatment for HAT. Spatial determinants for HAT are poorly understood across small areas. This study examines the relationships between Rhodesian HAT and several environmental, climatic and social factors in two newly affected districts, Kaberamaido and Dokolo. A one-step logistic regression analysis of HAT prevalence and a two-step logistic regression method permitted separate analysis of both HAT occurrence and HAT prevalence. Both the occurrence and prevalence of HAT were negatively correlated with distance to the closest livestock market in all models. The significance of distance to the closest livestock market strongly indicates that HAT may have been introduced to this previously unaffected area via the movement of infected, untreated livestock from endemic areas. This illustrates the importance of the animal reservoir in disease transmission, and highlights the need for trypanosomiasis control in livestock and the stringent implementation of regulations requiring the treatment of cattle prior to sale at livestock markets to prevent any further spread of Rhodesian HAT within Uganda. PMID- 20016847 TI - Discrepant prevalence and incidence of Leishmania infection between two neighboring villages in Central Mali based on Leishmanin skin test surveys. AB - Apart from a single report, the last publication of cutaneous leishmaniasis (CL) in Mali dates back more than 20 years. The absence of information on the current status of CL in Mali led us to conduct a cohort study in Kemena and Sougoula, two villages in Central Mali from which cases of CL have been recently diagnosed by Mali's reference dermatology center in Bamako. In May 2006, we determined the baseline prevalence of Leishmania infection in the two villages using the leishmanin skin test (LST). LST-negative individuals were then re-tested over two consecutive years to estimate the annual incidence of Leishmania infection. The prevalence of Leishmania infection was significantly higher in Kemena than in Sougoula (45.4% vs. 19.9%; OR: 3.36, CI: 2.66-4.18). The annual incidence of Leishmania infection was also significantly higher in Kemena (18.5% and 17% for 2007 and 2008, respectively) than in Sougoula (5.7% for both years). These data demonstrate that the risk of Leishmania infection was stable in both villages and confirm the initial observation of a significantly higher risk of infection in Kemena (OR: 3.78; CI: 2.45-6.18 in 2007; and OR: 3.36; CI: 1.95-5.8 in 2008; P<0.005). The absence of spatial clustering of LST-positive individuals in both villages indicated that transmission may be occurring anywhere within the villages. Although Kemena and Sougoula are only 5 km apart and share epidemiologic characteristics such as stable transmission and random distribution of LST-positive individuals, they differ markedly in the prevalence and annual incidence of Leishmania infection. Here we establish ongoing transmission of Leishmania in Kemena and Sougoula, Central Mali, and are currently investigating the underlying factors that may be responsible for the discrepant infection rates we observed between them. TRIAL REGISTRATION: ClinicalTrials.gov NCT00344084. PMID- 20016848 TI - Human probing behavior of Aedes aegypti when infected with a life-shortening strain of Wolbachia. AB - BACKGROUND: Mosquitoes are vectors of many serious pathogens in tropical and sub tropical countries. Current control strategies almost entirely rely upon insecticides, which increasingly face the problems of high cost, increasing mosquito resistance and negative effects on non-target organisms. Alternative strategies include the proposed use of inherited life-shortening agents, such as the Wolbachia bacterium. By shortening mosquito vector lifespan, Wolbachia could potentially reduce the vectorial capacity of mosquito populations. We have recently been able to stably transinfect Aedes aegypti mosquitoes with the life shortening Wolbachia strain wMelPop, and are assessing various aspects of its interaction with the mosquito host to determine its likely impact on pathogen transmission as well as its potential ability to invade A. aegypti populations. METHODOLOGY/PRINCIPAL FINDINGS: Here we have examined the probing behavior of Wolbachia-infected mosquitoes in an attempt to understand both the broader impact of Wolbachia infection on mosquito biology and, in particular, vectorial capacity. The probing behavior of wMelPop-infected mosquitoes at four adult ages was examined and compared to uninfected controls during video-recorded feeding trials on a human hand. Wolbachia-positive insects, from 15 days of age, showed a drastic increase in the time spent pre-probing and probing relative to uninfected controls. Two other important features for blood feeding, saliva volume and apyrase content of saliva, were also studied. CONCLUSIONS/SIGNIFICANCE: As A. aegypti infected with wMelPop age, they show increasing difficulty in completing the process of blood feeding effectively and efficiently. Wolbachia-infected mosquitoes on average produced smaller volumes of saliva that still contained the same amount of apyrase activity as uninfected mosquitoes. These effects on blood feeding behavior may reduce vectorial capacity and point to underlying physiological changes in Wolbachia-infected mosquitoes. PMID- 20016849 TI - The catalytic subunit of protein phosphatase 1 gamma regulates thrombin-induced murine platelet alpha(IIb)beta(3) function. AB - BACKGROUND: Hemostasis and thrombosis are regulated by agonist-induced activation of platelet integrin alpha(IIb)beta(3). Integrin activation, in turn is mediated by cellular signaling via protein kinases and protein phosphatases. Although the catalytic subunit of protein phosphatase 1 (PP1c) interacts with alpha(IIb)beta(3), the role of PP1c in platelet reactivity is unclear. METHODOLOGY/PRINCIPAL FINDINGS: Using gamma isoform of PP1c deficient mice (PP1cgamma(-/-)), we show that the platelets have moderately decreased soluble fibrinogen binding and aggregation to low concentrations of thrombin or protease activated receptor 4 (PAR4)-activating peptide but not to adenosine diphosphate (ADP), collagen or collagen-related peptide (CRP). Thrombin-stimulated PP1cgamma( /-) platelets showed decreased alpha(IIb)beta(3) activation despite comparable levels of alpha(IIb)beta(3), PAR3, PAR4 expression and normal granule secretion. Functions regulated by outside-in integrin alpha(IIb)beta(3) signaling like adhesion to immobilized fibrinogen and clot retraction were not altered in PP1cgamma(-/-) platelets. Thrombus formation induced by a light/dye injury in the cremaster muscle venules was significantly delayed in PP1cgamma(-/-) mice. Phosphorylation of glycogen synthase kinase (GSK3)beta-serine 9 that promotes platelet function, was reduced in thrombin-stimulated PP1cgamma(-/-) platelets by an AKT independent mechanism. Inhibition of GSK3beta partially abolished the difference in fibrinogen binding between thrombin-stimulated wild type and PP1cgamma(-/-) platelets. CONCLUSIONS/SIGNIFICANCE: These studies illustrate a role for PP1cgamma in maintaining GSK3beta-serine9 phosphorylation downstream of thrombin signaling and promoting thrombus formation via fibrinogen binding and platelet aggregation. PMID- 20016850 TI - Clinical and financial outcomes due to methicillin resistant Staphylococcus aureus surgical site infection: a multi-center matched outcomes study. AB - BACKGROUND: The clinical and financial outcomes of SSIs directly attributable to MRSA and methicillin-resistance are largely uncharacterized. Previously published data have provided conflicting conclusions. METHODOLOGY: We conducted a multi center matched outcomes study of 659 surgical patients. Patients with SSI due to MRSA were compared with two groups: matched uninfected control patients and patients with SSI due to MSSA. Four outcomes were analyzed for the 90-day period following diagnosis of the SSI: mortality, readmission, duration of hospitalization, and hospital charges. Attributable outcomes were determined by logistic and linear regression. PRINCIPAL FINDINGS: In total, 150 patients with SSI due to MRSA were compared to 231 uninfected controls and 128 patients with SSI due to MSSA. SSI due to MRSA was independently predictive of readmission within 90 days (OR = 35.0, 95% CI 17.3-70.7), death within 90 days (OR = 7.27, 95% CI 2.83-18.7), and led to 23 days (95% CI 19.7-26.3) of additional hospitalization and $61,681 (95% 23,352-100,011) of additional charges compared with uninfected controls. Methicillin-resistance was not independently associated with increased mortality (OR = 1.72, 95% CI 0.70-4.20) nor likelihood of readmission (OR = 0.43, 95% CI 0.21-0.89) but was associated with 5.5 days (95% CI 1.97-9.11) of additional hospitalization and $24,113 (95% 4,521-43,704) of additional charges. CONCLUSIONS/SIGNIFICANCE: The attributable impact of S. aureus and methicillin-resistance on outcomes of surgical patients is substantial. Preventing a single case of SSI due to MRSA can save hospitals as much as $60,000. PMID- 20016851 TI - Deletion of RAGE causes hyperactivity and increased sensitivity to auditory stimuli in mice. AB - The receptor for advanced glycation end-products (RAGE) is a multi-ligand receptor that belongs to the immunoglobulin superfamily of cell surface receptors. In diabetes and Alzheimer's disease, pathological progression is accelerated by activation of RAGE. However, how RAGE influences gross behavioral activity patterns in basal condition has not been addressed to date. In search for a functional role of RAGE in normal mice, a series of standard behavioral tests were performed on adult RAGE knockout (KO) mice. We observed a solid increase of home cage activity in RAGE KO. In addition, auditory startle response assessment resulted in a higher sensitivity to auditory signal and increased prepulse inhibition in KO mice. There were no significant differences between KO and wild types in behavioral tests for spatial memory and anxiety, as tested by Morris water maze, classical fear conditioning, and elevated plus maze. Our results raise a possibility that systemic therapeutic treatments to occlude RAGE activation may have adverse effects on general activity levels or sensitivity to auditory stimuli. PMID- 20016852 TI - Genetic variation of the human urinary tract innate immune response and asymptomatic bacteriuria in women. AB - BACKGROUND: Although several studies suggest that genetic factors are associated with human UTI susceptibility, the role of DNA variation in regulating early in vivo urine inflammatory responses has not been fully examined. We examined whether candidate gene polymorphisms were associated with altered urine inflammatory profiles in asymptomatic women with or without bacteriuria. METHODOLOGY: We conducted a cross-sectional analysis of asymptomatic bacteriuria (ASB) in 1,261 asymptomatic women ages 18-49 years originally enrolled as participants in a population-based case-control study of recurrent UTI and pyelonephritis. We genotyped polymorphisms in CXCR1, CXCR2, TLR1, TLR2, TLR4, TLR5, and TIRAP in women with and without ASB. We collected urine samples and measured levels of uropathogenic bacteria, neutrophils, and chemokines. PRINCIPAL FINDINGS: Polymorphism TLR2_G2258A, a variant associated with decreased lipopeptide-induced signaling, was associated with increased ASB risk (odds ratio 3.44, 95%CI; 1.65-7.17). Three CXCR1 polymorphisms were associated with ASB caused by gram-positive organisms. ASB was associated with urinary CXCL-8 levels, but not CXCL-5, CXCL-6, or sICAM-1 (P< or =0.0001). Urinary levels of CXCL-8 and CXCL-6, but not ICAM-1, were associated with higher neutrophil levels (P< or =0.0001). In addition, polymorphism CXCR1_G827C was associated with increased CXCL-8 levels in women with ASB (P = 0.004). CONCLUSIONS: TLR2 and CXCR1 polymorphisms were associated with ASB and a CXCR1 variant was associated with urine CXCL-8 levels. These results suggest that genetic factors are associated with early in vivo human bladder immune responses prior to the development of symptomatic UTIs. PMID- 20016854 TI - Diabodies: molecular engineering and therapeutic applications. AB - Bispecific antibodies are capable of interacting with two different antigens and when selected properly, can redirect cytotoxic effector cells to tumor cells for effective killing. These antibodies are therefore of great interest in the research and development of cancer treatment. Over the last two decades, many different bispecific antibody-derived molecular formats have been described, among which diabodies represent an important class of engineered molecules that possess tumor-targeting function. Since diabodies were first introduced in the early 1990s, extensive efforts have been made to optimize their physicochemical and key functional properties, as well as to provide in vivo proof of concept of their antitumor efficacy in animal models. With the clinical validation of the T cell-retargeting mechanism for cancer therapy currently in place, there is renewed interest in this bispecific class of biologic molecules, with additional novel formats being described in recent years. Even with the remaining challenges of the manufacturing yields and drug-like properties, diabodies and their derivatives remain viable therapeutic modalities that warrant further consideration and development. PMID- 20016853 TI - Dendritic cell modulation as a new interventional approach for the treatment of asthma. AB - Dendritic cells (DCs) have a central role in the immune system, as they control the adaptive immune response and mediate both protective immunity and the maintenance of immune tolerance to self antigens. In this review, we will summarize the recent advances regarding the subsets of DCs and how they regulate the differentiation of naive CD4(+) T cells towards different populations of T helper cells. We will particularly describe the role of DCs in the development and regulation of allergic diseases and asthma, and discuss the capacity of DCs to induce proallergenic Th2 cells versus regulatory T cells. Undoubtedly, tolerogenic DCs play a crucial role in the induction of regulatory cells. Understanding the biology of these cells will help us design novel strategies to cure or prevent allergic diseases and asthma. PMID- 20016855 TI - Receptorome screening: a powerful, facile approach to better understand and engineer drugs. AB - Receptorome screening is the process of characterizing one or more compounds for pharmacological activity (e.g., inhibition of radioligand binding, positive or negative efficacy) at a large panel of 'targets' (e.g., biologically relevant recombinant G protein-coupled receptors, ion channels and small-molecule transporters). Recently, receptorome profiles have led to mechanistic insights into the actions -both intentional and adverse- of several drugs. In the present review, I discuss in detail how receptorome screening increased our understanding of three drugs (salvinorin A, amisulpride and fenfluramine) and a class of medications (atypical antipsychotics). The cases presented suggest that receptorome screening of current medications, as well as investigational drugs and future compounds destined for use in humans, might enable us to predict salutary effects, as well as side effects, at the preclinical stage, which would have important economic and public health implications. PMID- 20016856 TI - Beta-arrestin signaling complex as a target for antidepressants and as a depression marker. AB - Beta-arrestins uncouple G protein-coupled receptors (GPCRs) from G proteins and promote their internalization, leading to desensitization and downregulation and serving as negative regulators of GPCR signaling. beta-Arrestins also function as scaffold proteins, interacting with several cytoplasmic proteins and linking GPCRs to intracellular signaling pathways such as the mitogen-activated protein kinase (MAPK) cascade. Recent work has also revealed that beta-arrestins translocate from the cytoplasm to the nucleus and associate with transcription factors such as histone acetyltransferase p300 and cyclic adenosine monophosphate (cAMP)-responsive element-binding protein. These substances also interact with regulators of transcription factors. We review findings on the effects of antidepressants on beta-arrestins and the plethora of antidepressant effects on signal transduction elements in which beta-arrestins serve as signaling scaffold proteins, focusing on the three major groups of MAPKs: extracellular signal regulated kinases, c-Jun N-terminal kinases and p38 MAPKs, and on transcription factors and cofactors of which beta-arrestins mediate transcription regulation. PMID- 20016858 TI - Molecule of the month. XOMA-052. PMID- 20016857 TI - Magnetic resonance imaging biomarkers of ischemic stroke: criteria for the validation of primary imaging biomarkers. AB - Ischemic stroke is associated with a high rate of disability and death. Establishing valid biomarkers could help accelerate the approval of promising new therapies for stroke. Whereas many serum biomarkers have been evaluated, possible imaging biomarkers of stroke lack validation. Magnetic resonance imaging (MRI) is a very sensitive technique to study acute stroke and MRI parameters have been established to assess the outcome of acute stroke. This review reassesses the criteria for the validation of MRI biomarkers of acute ischemic stroke (MRI-BAS). Seven criteria were used to review the validity of the main MRI-BAS: vascular status, lesion volume, reversibility on diffusion-weighted imaging, perfusion alteration, penumbra studied with diffusion-perfusion mismatch, clinical diffusion mismatch, diffusion-angiography mismatch and hemorrhagic transformation. We analyzed the definitions of these biomarkers and the extent to which each fulfills the criteria for validation and found that few MRI-BAS have been fully validated. Further studies should help to improve the validation of current MRI-BAS and develop new biomarkers. PMID- 20016859 TI - Chronicles in drug discovery. PMID- 20016860 TI - An overview on stem cell research: a report on the 7th Annual Meeting of the International Society for Stem Cell Research. AB - The focus of the seventh annual meeting of the International Society for Stem Cell Research was placed on several topics, such as cancer stem cells, stem cell transplantation, tissue regeneration, biology of embryonic and adult stem cells of various species and production of induced pluripotent stem cells via reprogramming of mammalian somatic cells. This report provides an overview of stem cell research, based on the proceedings of the meeting. PMID- 20016866 TI - Quality of life of asthmatic adolescents: assessment of asthma severity, comorbidity, and life style. AB - OBJECTIVE: To assess the quality of life (QoL) of asthmatic adolescents and its association with asthma severity, chronic diseases, and life style. METHOD: Cross sectional study involving 210 female and male asthmatic adolescents between 12 and 21 years old. The Paediatric Asthma Quality of Life Questionnaire, a self administered instrument, was used. Explanatory variables were: allergic diseases, use of medication, passive smoking, occupational status, asthma severity, and life style. The analyses considered the dichotomous outcome (good/poor QoL) based on the mean scores. Generalized linear models (log-binomial) were used to calculate the crude and adjusted prevalence ratios (PR). RESULTS: Forty-six per cent of the girls and 57% of the boys had poor QoL. There was no correlation between chronic diseases and poor QoL. Low educational level, use of medication, passive smoking, and occupational factors had a statistically significant association (p < 0.05) with poor QoL. The adjusted analysis showed that severe asthma (PR = 1.53; 95%CI 1.12-2.11), use of medication (PR = 1.58; 95%CI 1.09 2.28), having asthma diagnosis established less than 5 years ago (PR = 1.30; 95%CI 0.97-1.86), passive smoking (PR = 1.38; 95%CI 1.35-2.00), and being employed (PR = 1.30; 95%CI 0.96-1.74) were associated with poor QoL. CONCLUSION: A multidisciplinary team needs to face the challenge of providing good QoL with the purpose of making these patients better adapted to society and to their own needs. PMID- 20016867 TI - Pacifier habit: history and multidisciplinary view. AB - OBJECTIVES: To review the history of pacifiers and to compile a multidisciplinary literature review, searching for pros and cons with the purpose of providing health professionals with arguments when parents request guidance. SOURCES: History and art books, as well as non-medical literature and museums were used in the historical survey. Multidisciplinary data were collected from MEDLINE, LILACS, SciELO, and The Cochrane Library. Search criteria were: the keyword "pacifiers" present in articles published in the last 5 years that included abstract and were written in Portuguese, English, or Spanish. SUMMARY OF THE FINDINGS: There is evidence that their precursors have been used since the Neolithic Period to calm down children. Small balls made of fabric containing food were portrayed in paintings. Other balls made of non-perishable material persisted throughout time. Pacifiers have been used to stimulate sucking or to coordinate this reflex, promoting an earlier beginning of the oral feeding of newborns. Some authors suggest that pacifiers reduce the incidence of the sudden death syndrome, but the topic is controversial. Pacifiers prevent the establishment of breastfeeding and lead to weaning. Their use may cause suffocation, poisoning, or allergies and increases the risk of caries, infections, and intestinal parasitic diseases. Harmful effects are related to frequency, duration, and intensity of the habit. It should be discontinued by the age of 3 or 4 in order not to affect speech and dentition. CONCLUSIONS: There are more harmful effects than benefits. It is advisable that health professionals inform parents of the pros and cons of pacifiers so that they can make a conscious decision regarding its use. PMID- 20016868 TI - Stroke in children. AB - OBJECTIVES: To summarize 1) the definitions and epidemiological features of stroke in children; 2) the main risk factors that can lead to stroke in pediatrics and neonatology; 3) the main pathophysiological features involved in the genesis of brain injury in stroke; 4) the clinical manifestations and imaging diagnosis; and 5) the latest recommendations concerning the support measures, treatment, and prophylaxis of stroke in children. SOURCES: A review of the literature published in PubMed, EMBASE, and SciELO databases using the search terms stroke, pediatrics, and neonatology was performed, including relevant references from the chosen texts. SUMMARY OF THE FINDINGS: Stroke in children are rare conditions, with incidence rates among two and eight per 100,000 children up to 14 years, and most cases show an underlying disease such as heart diseases, prothrombotic conditions, sickle-cell disease, and vascular malformations. There are no specific guidelines currently in place for the treatment of stroke in children, although central elements include support treatment, monitoring, and anticoagulation as secondary prevention in certain cases. Prognosis depends on the extent of brain damage and the underlying disease but recurrence rates are high in most cases. CONCLUSIONS: Early diagnosis of stroke in children is very important and pediatricians should be aware of the lack of specificity of the symptoms to avoid late sequelae and improve life quality. PMID- 20016869 TI - High prevalence of children colonized with penicillin-resistant Streptococcus pneumoniae in public day-care centers. AB - OBJECTIVES: To investigate the prevalence of Streptococcus pneumoniae (pneumococci) in the nasopharynx of healthy children enrolled in public day-care centers of the municipality of Umuarama, state of Parana, Brazil. The susceptibility of the pneumococcal strains to antimicrobial agents was also studied. METHODS: Nasopharyngeal specimens from 212 children were collected from April to October 2008. After the specimens were seeded in blood agar and incubated at 37 degrees C for 24-48 hours, the colonies suspected of belonging to S. pneumoniae were identified using alpha-hemolysis, optochin sensitivity, and bile solubility test. Penicillin susceptibility was investigated using the disk diffusion and dilution tests. Susceptibility to the other antimicrobial agents indicated for the treatment of pneumococcal infections was investigated using the disk diffusion test. RESULTS: The prevalence of nasopharyngeal pneumococci was 43.4% (92/212), with higher rates in children between 2 and 5 years old (p = 0.0005). There was no significant difference between sexes. Intermediate and full resistance to penicillin were found in 34.8 (32/92) and 22.8% (21/92) isolates, respectively. Sixty-seven strains (72.8%) were resistant to sulfamethoxazole trimethoprim, eight (8.7%) were resistant to erythromycin, and six (6.5%) to tetracycline. One strain was resistant to clindamycin (1.1%) and another was resistant to chloramphenicol (1.1%). All strains were sensitive to levofloxacin, ofloxacin, rifampicin, telithromycin, linezolid, and vancomycin. Nine strains were considered multiresistant because they were resistant to three or more classes of antimicrobial agents. CONCLUSIONS: The present study detected a high prevalence of healthy children colonized with penicillin-resistant S. pneumoniae strains who may be important reservoirs of this pathogen in the community. PMID- 20016870 TI - Oral vs. intravenous empirical antimicrobial therapy in febrile neutropenic patients receiving childhood cancer chemotherapy. AB - OBJECTIVE: To compare the use of intravenous vs. oral antibiotic therapy. METHODS: All febrile neutropenic patients younger than 18 years old with low risk of complications and receiving chemotherapy were selected. The study was conducted from 2002 to 2005 at the Pediatric Oncology Unit of Hospital de Clinicas de Porto Alegre, Porto Alegre, Brazil. Patients were divided into group A and group B and were randomly assigned to receive oral or intravenous therapy. The empirical antimicrobial treatment used for group A consisted in oral ciprofloxacin plus amoxicillin-clavulanate and intravenous placebo, and group B received cefepime and oral placebo. RESULTS: A total of 91 consecutive episodes of febrile neutropenia in 58 children were included in the study. For patients of group A, treatment failure rate was 51.2%; the mean length of hospital stay was 8 days (range 2-10 days). For patients treated with intravenous antibiotic therapy, treatment failure rate was 45.8%; the mean length of hospital stay was 7 days (range 3-10 days). CONCLUSION: There was no difference in the outcome in oral vs. intravenous therapy. There is need of larger randomized trials before oral empirical therapy administered to this population should be considered the new standard of treatment. PMID- 20016871 TI - Nutritional quality and food expenditure in preschool children. AB - OBJECTIVE: To assess correlations between the cost and the nutritional quality of the diets of preschool children from low socioeconomic status families, taking into account intakes of micronutrients and foods with high concentrations of sugars and fats. METHODS: This was a cross-sectional study undertaken with 346 children aged 3 to 4 years recruited for the "Ten steps in action" (BRATSA I) study, who comprise a nested cohort from the randomized field study. Two 24-hour dietary recall surveys were conducted. Expenditure on food was calculated by taking the price of each food, as verified at a number of different establishments, and adjusting it for the quantity eaten. RESULTS: Mean expenditure on food for one child was R$ 100.17+/-34.1 per month. There was a positive correlation between intakes of iron (r = 0.115; p = 0.033), zinc (r = 0.214; p < 0.001), and vitamins A (r = 0.197; p < 0.001) and C (r = 0.162; p < 0.001), adjusted to 1,000 kcal, and expenditure on food/1,000 kcal. There were no significant relationships between expenditure on food/1,000 kcal and risk of overweight (p = 0.208) or intake of foods with a high fat or sugar content (p = 0.894 and p = 0.964). CONCLUSIONS: The study found that consumption of energy provided by fat and sugar-rich foods was not associated with expenditure on feeding these 3-to-4-year-olds. In contrast, nutritional quality, assessed in the form of essential micronutrient intakes, demonstrated a positive correlation with food costs. PMID- 20016872 TI - Impact of divorce on the quality of life in school-age children. AB - OBJECTIVE: To assess psychosocial quality of life in school-age children of divorced parents. METHODS: A cross-sectional survey was conducted at the pediatric outpatient clinic of a community hospital. Children 5 to 12 years old from married families and divorced families were included. Child quality of life was assessed through maternal reports using a Child Health Questionnaire-Parent Form 50. A multiple linear regression model was constructed including clinically relevant variables significant on univariate analysis (beta coefficient and 95%CI). RESULTS: Three hundred and thirty families were invited to participate and 313 completed the questionnaire. Univariate analysis showed that quality of life was significantly associated with parental separation, child sex, time spent with the father, standard of living, and maternal education. In a multiple linear regression model, quality of life scores decreased in boys -4.5 (-6.8 to -2.3) and increased for time spent with the father 0.09 (0.01 to 0.2). In divorced families, multiple linear regression showed that quality of life scores increased when parents had separated by mutual agreement 6.1 (2.7 to 9.4), when the mother had university level education 5.9 (1.7 to 10.1) and for each year elapsed since separation 0.6 (0.2 to 1.1), whereas scores decreased in boys -5.4 (-9.5 to -1.3) and for each one-year increment of maternal age -0.4 (-0.7 to -0.05). CONCLUSION: Children's psychosocial quality of life was affected by divorce. The Child Health Questionnaire can be useful to detect a decline in the psychosocial quality of life. PMID- 20016873 TI - Hospitalization of children for asthma in the Brazilian Amazon: trend and spatial distribution. AB - OBJECTIVE: To analyze the distribution of hospitalizations of children for asthma in the states comprising the Brazilian Amazon, as well as the variations of hospitalization according to climatic seasonality. METHODS: A descriptive study of the distribution of hospitalizations for asthma of individuals aged 0 to 14 years living in the Brazilian Amazon, according to trend, spatial distribution, and climatic seasonality over the period from 2001 to 2007. We used the database of authorizations for hospitalizations (AIH) of the Hospital Information System (SIH/SUS) of the Brazilian Ministry of Health. RESULTS: The spatial distribution of hospitalizations for asthma resembles the configuration of the "arc of deforestation," mainly in the states of Rondonia and Maranhao. The distribution of annual hospitalizations for asthma decreased from 8.1 to 2.6 hospitalizations/1,000 inhabitants during the period. There was a peak of hospitalizations in March and May in all states. The seasonal differences were on average 10%, with the highest rates during the rainy season. CONCLUSION: We concluded that hospitalizations for asthma are more frequent in the months of more intense rainfall, with greater magnitude in the states comprising the "arc of deforestation" in the Brazilian Amazon, especially Rondonia and Maranhao. PMID- 20016874 TI - Glycemic control in Brazilian youth with type 1 diabetes. PMID- 20016875 TI - Amelioration of hyperglycemia by intestinal overexpression of glucagon-like peptide-1 in mice. AB - To investigate whether the local production of glucagon-like peptide-1 (GLP-1) in the intestine can differentiate intestinal stem/progenitor cells into insulin producing cells, we intra-intestinally injected a recombinant adenovirus expressing GLP-1 (rAd-GLP-1) into diabetic mice. There were no significant differences in body weight or food intake between rAd-GLP-1- and rAd-betaGAL treated control mice. rAd-GLP-1-treated mice showed intestinal insulin mRNA expression, insulin- and glucagon-positive cells in the intestine, and significantly increased serum insulin, but not glucagon. rAd-GLP-1 injection significantly reduced blood glucose levels and improved glucose tolerance compared with controls. Expression of transcription factors related to beta cell differentiation, neurogenin 3 (ngn3) and neurogenin differentiation factor (NeuroD), was detected in the intestine at 2 weeks after rAd-GLP-1 injection. We suggest that expression of GLP-1 in the intestine by intra-intestinal delivery of rAd-GLP-1 may induce differentiation of intestinal stem/progenitor cells into insulin-producing cells, mediated by ngn3 and NeuroD expression, contributing to lowered blood glucose levels in diabetic mice. PMID- 20016876 TI - The environmental challenge for analytical sciences. AB - In this paper the major elements of the European Union's policy on environmental protection and sustainable development and the resulting challenges for analytical sciences are presented. The priority issues dealt with are: Sustainable management of natural resources: air, water and soil Climate change and clean energy Global development cooperation Analytical sciences are required to provide policy-relevant information for the development and implementation of European Union legislation and form a strong pillar for a sustainable evolution of our region and our planet. It shows what information needs to be provided, how the necessary quality levels can be achieved and what new approaches, e.g. combining measurements and modelling, or earth observations with in situ chemical/physical measurements, need to be taken to achieve an integrated assessment of the state of the environment and to develop approaches for sustainable development. PMID- 20016877 TI - Study on elution behavior of poly(amidoamine) dendrimers and their interaction with bovine serum albumin in asymmetrical flow field-flow fractionation. AB - Polyamidoamine (PAMAM) dendrimers have an amine surface and an ethylenediamine core and are of great interest in various applications such as in drug delivery. Physiochemical properties of PAMAM dendrimers vary with pH. At neutral to basic pH, PAMAM dendrimers are either weakly charged or uncharged and tend to adsorb on to the neutral packing material, making chromatographic separation of the dendrimers difficult. Asymmetrical flow field-flow fractionation (AsFlFFF) was tested as an alternative to the chromatographic techniques for separation of the PAMAM dendrimers. AsFlFFF provided generation-based separation of the dendrimers even at neutral and basic pH. The elution time increased gradually as the generation number (and thus the size) increased. Separation of impurities such as generational or missing-arm impurities and aggregates from the main population was also achieved. Electrostatic and hydrophobic interactions (e.g., repulsive elecrostatic interaction among the dendrimer molecules or attractive hydrophobic interaction between the dendrimer molecules and the membrane) may result in an inaccurate size measurement. Careful optimization of experimental conditions such as the flow rate, pH, and the salt concentration may be required to minimize the interactions with the membrane. AsFlFFF was also tested for a study on the interaction between the PAMAM dendrimers and proteins. AsFlFFF was able to show the growth in the size of bovine serum albumin (BSA) when BSA is mixed with increasing amounts of PAMAM dendrimers. Results suggest that, with proper optimization, AsFlFFF could become a useful tool for separation and characterization of large charged molecules such as PAMAM dendrimers. PMID- 20016878 TI - A rapid and universal bacteria-counting approach using CdSe/ZnS/SiO2 composite nanoparticles as fluorescence probe. AB - In this paper, a rapid, simple, and sensitive method was described for detection of the total bacterial count using SiO(2)-coated CdSe/ZnS quantum dots (QDs) as a fluorescence marker that covalently coupled with bacteria using glutaraldehyde as the crosslinker. Highly luminescent CdSe/ZnS were prepared by applying cadmium oxide and zinc stearate as precursors instead of pyrophoric organometallic precursors. A reverse-microemulsion technique was used to synthesize CdSe/ZnS/SiO(2) composite nanoparticles with a SiO(2) surface coating. Our results showed that CdSe/ZnS/SiO(2) composite nanoparticles prepared with this method possessed highly luminescent, biologically functional, and monodispersive characteristics, and could successfully be covalently conjugated with the bacteria. As a demonstration, it was found that the method had higher sensitivity and could count bacteria in 3 x 10(2) CFU/mL, lower than the conventional plate counting and organic dye-based method. A linear relationship of the fluorescence peak intensity (Y) and the total bacterial count (X) was established in the range of 3 x 10(2)-10(7) CFU/mL using the equation Y = 374.82X-938.27 (R = 0.99574). The results of the determination for the total count of bacteria in seven real samples were identical with the conventional plate count method, and the standard deviation was satisfactory. PMID- 20016879 TI - Use of pJANUS-02-001 as a calibrator plasmid for Roundup Ready soybean event GTS 40-3-2 detection: an interlaboratory trial assessment. AB - Owing to the labelling requirements of food and feed products containing materials derived from genetically modified organisms, quantitative detection methods have to be developed for this purpose, including the necessary certified reference materials and calibrator standards. To date, for most genetically modified organisms authorized in the European Union, certified reference materials derived from seed powders are being developed. Here, an assessment has been made on the feasibility of using plasmid DNA as an alternative calibrator for the quantitative detection of genetically modified organisms. For this, a dual-target plasmid, designated as pJANUS-02-001, comprising part of a junction region of genetically modified soybean event GTS-40-3-2 and the endogenous soybean-specific lectin gene was constructed. The dynamic range, efficiency and limit of detection for the soybean event GTS-40-3-2 real-time quantitative polymerase chain reaction (Q-PCR) system described by Terry et al. (J AOAC Int 85(4):938-944, 2002) were shown to be similar for in house produced homozygous genomic DNA from leaf tissue of soybean event GTS-40-3-2 and for plasmid pJANUS 02-001 DNA backgrounds. The performance of this real-time Q-PCR system using both types of DNA templates as calibrator standards in quantitative DNA analysis was further assessed in an interlaboratory trial. Statistical analysis and fuzzy logic-based interpretation were performed on critical method parameters (as defined by the European Network of GMO Laboratories and the Community Reference Laboratory for GM Food and Feed guidelines) and demonstrated that the plasmid pJANUS-02-001 DNA represents a valuable alternative to genomic DNA as a calibrator for the quantification of soybean event GTS-40-3-2 in food and feed products. PMID- 20016880 TI - Identification of biochemical changes in lactovegetarian urine using 1H NMR spectroscopy and pattern recognition. AB - A vegetarian diet has been demonstrated to have a profound influence on human metabolism as well as to aid the prevention of several chronic diseases relative to an omnivorous diet. However, there have been no systematic metabolomic studies on all of the biochemical changes induced in human subjects by long-term vegetarianism. In this study, (1)H NMR spectroscopy in combination with multivariate statistical analysis was applied to explore the variability in the metabolic urinary profiles of healthy populations from four groups: lactovegetarian male (VEGMALE), lactovegetarian female (VEGFEMALE), omnivorous male (OMNMALE), and omnivorous female (OMNFEMALE). Differences in metabolic profiles were examined in relation to diet and gender by principal component analysis (PCA) and spectral integrals. It was found that the most influential low molecular weight metabolites responsible for the differences between the diet groups were N-acetyl glycoprotein (NAG), succinate, citrate, trimethylamine-N oxide (TMAO), taurine, glycine, hippurate, phenylalanine, methylhistidine and formate, whereas for the differences in gender groups the most discriminatory metabolites were NAG, succinate, creatinine, arginine, TMAO, taurine, hippurate, mannitol, phenylalanine, and methylhistidine. The results from the PCA of all four groups indicated that diet plays a greater role in influencing metabolite differences than gender. As an exploration, this work shows the potential of metabolomics when applied to nutritional and physiological studies, and it will aid further studies. PMID- 20016881 TI - Applications of TGA in quality control of SWCNTs. AB - Carbon nanotubes exhibit a range of chemistries, including mixtures of different nanotube diameters, lengths, and chiralities coupled with various concentrations of metallic and non-nanotube-carbon impurities. The performance of a given material for a specific application depends on the chemistry, which is dictated in large part by the manufacturing process. Here, thermogravimetric analysis is utilized as a bulk characterization method for determining nanotube quality after manufacturing. The application of thermogravimetric analysis for quantifying basic nanotube chemistry is described (e.g., carbon-to-metal content, homogeneity). In addition, extension of the method to analyze specific nanotube properties (i.e., length and diameter) is reported. Results indicate that thermogravimetric analysis is sufficiently sensitive to enable quality control at both the macro-scale (carbon-to-metal ratio) and nano-scale (single-walled to multi-walled) and can detect subtle modifications in manufacturing processes. PMID- 20016882 TI - Use of the quartz crystal microbalance to monitor ligand-induced conformational rearrangements in HIV-1 envelope protein gp120. AB - We evaluated the potential of a quartz crystal microbalance with dissipation monitoring (QCM-D) to provide a sensitive, label-free method for detecting the conformational rearrangement of glycoprotein gp120 upon binding to different ligands. This glycoprotein is normally found on the envelope of the HIV-1 virus and is involved in viral entry into host cells. It was immobilized on the surface of the sensing element of the QCM-D and was exposed to individual solutions of several different small-molecule inhibitors as well as to a solution of a soluble form of the host cell receptor to which gp120 binds. Instrument responses to ligand-triggered changes were in qualitative agreement with conformational changes as suggested by other biophysical methods. PMID- 20016883 TI - Characterization of gold nanorods in vivo by integrated analytical techniques: their uptake, retention, and chemical forms. AB - Integrated analytical techniques were used to study the tissue distribution and structural information of gold nanorods (Au NRs) in Sprague-Dawley rats through tail intravenous injection. Before in vivo experiments were conducted, careful characterization of Au NRs was performed. The zeta potential proved that adsorption of bovine serum albumin on Au NRs turned the surface charges from positive to negative as in an in vitro simulation. The biodistribution of Au NRs was investigated quantitatively by inductively coupled plasma mass spectrometry at different time points after injection. As target tissues, both liver and spleen were chosen to further demonstrate the intracellular localization of Au NRs by the combination of transmission electron microscopy and energy-dispersive X-ray spectroscopy. Moreover, synchrotron-radiation-based X-ray absorption spectroscopy was employed and it was observed that long-term retention of Au NRs in liver and spleen did not induce obvious changes in the oxidation states of gold. Therefore, the present systematic method can provide important information about the fates of Au NRs in vivo and can also be extended to study the biological effects of other metallic nanomaterials in the future. PMID- 20016884 TI - Imaging techniques for acute ischemic stroke: nice gadgets or essential tools for effective treatment? PMID- 20016885 TI - Complications and outcomes following extracorporeal shock wave lithotripsy: a prospective study of 3,241 patients. AB - Extracorporeal shock wave lithotripsy (SWL) has become the least invasive treatment modality with high success rates for urinary calculi; however, its established efficacy has been associated with a number of side effects and complications. This study sought to further evaluate the incidence rate and management of the post-SWL complications and also the efficiency of procedure in a large scale of patients. During a 51-month period, 3,241 consecutive adult patients with the mean age of 38.1 years (range 15-75) and urinary calculi (>or=4 mm) underwent SWL at our referral center and were followed for 3 months prospectively. Overall, 3,614 stones [kidneys (83.5%), ureters (15.8%) and bladder (0.7%)] in 3,241 patients were treated requiring 7,245 SWL sessions. Stone-free state occurred in 71.5% calculi and success rate in 79.8% patients. The re-treatment was necessary in 37.2% patients. Auxiliary procedure and efficiency quotient were 5.6% and 0.50, respectively. SWL success rate decreased as the stone size increased (P < 0.0001). The stone-free rate was correlated with the location of the stone. During the study period, 4,075 complications occurred in our patients. Colicky pain (40%) was the most frequent symptom followed by gross hematuria (32%) and steinstrasse (24.2%). Symptomatic bacteriuria developed in 9.7% patients; Escherichia coli (30.4%) was the most causative organism. In conclusion, the complication rate following SWL was high in our study; however, the majority was mild and managed conservatively or with the minimal intervention. Moreover, the management of urinary calculi in adults using SWL was proved to be safe and efficient, particularly for ureteral stones <10 mm, renal pelvic stones <20 mm, and bladder stones <30 mm. PMID- 20016886 TI - Current and historically used pesticides in residential soil from 11 homes in Atlanta, Georgia, USA. AB - We used a multiresidue, gas chromatography/mass spectrometry-based method to measure seven pyrethroid, five organophosphorus (OP), and six organochlorine pesticides in soil collected from 11 Atlanta homes in 2006. Our objective was to collect preliminary data for a larger study of pesticide exposures among Atlanta children. The pyrethroid insecticides (cis- and trans-permethrin, bioallethrin) were the most commonly detected analytes, giving evidence of widespread outdoor use among our study homes. Our pyrethroid insecticide detection frequencies were higher than those reported in a recent study of Ohio and North Carolina homes; however, our maximum values were approximately half of those reported. We detected the target OP pesticides in only a few samples, but we found two restricted-use OP pesticides--methyl parathion and terbufos--and thus possible evidence of illegal residential use or environmental persistence in soil. We also detected dichlorodiphenyltrichloroethane (DDT) and dichlorodiphenyldichloroethane (DDE) in samples from six homes. Although our small sample size limits comparison to other studies, our results provide evidence that residential soil is a potential source of human exposure to both current and historically used pesticides. PMID- 20016887 TI - Cd, Pb and Zn oral bioaccessibility of urban soils contaminated in the past by atmospheric emissions from two lead and zinc smelters. AB - Ingestion of dust or soil particles could pose a potential health risk due to long-term metal trace element (MTE) exposure. Twenty-seven urban topsoil samples (kitchen garden and lawn) were collected and analyzed for Cd, Pb and Zn using the unified Bioaccessibility Research Group of Europe (BARGE) method (UBM) test to estimate the human bioaccessibility of these elements. The quantities of Cd, Pb and Zn extracted from soils indicated, on average, 68, 62 and 47% bioaccessibility, respectively, in the gastric phase and 31, 32 and 23% bioaccessibility, respectively, in the gastro-intestinal phase. Significant positive correlations were observed between concentrations extracted with UBM and total MTE contents. Stepwise multiple regression analysis showed that human bioaccessibility was also affected by some physico-chemical soil parameters (i.e. total nitrogen, carbonates, clay contents and pH). The unified test presents some valuable data for risk assessment. Indeed, the incorporation of oral bioaccessible concentrations into risk estimations could give more realistic information for health risk assessment. PMID- 20016888 TI - Articular cartilage and labral lesions of the glenohumeral joint: diagnostic performance of 3D water-excitation true FISP MR arthrography. AB - OBJECTIVE: To evaluate the diagnostic performance of MR arthrography in the detection of articular cartilage and labral lesions of the glenohumeral joint using a transverse 3D water-excitation true fast imaging with steady-state precession (FISP) sequence. MATERIALS AND METHODS: Seventy-five shoulders were included retrospectively. Shoulder arthroscopy was performed within 6 months of MR arthrography. MR images were evaluated separately by two radiologists. They were blinded to clinical and arthroscopic information. Arthroscopy served as the reference standard. RESULTS: For the detection of humeral cartilage lesions, sensitivities and specificities were 86% (12/14)/89% (50/56) for observer 1 and 93%/86% for observer 2) for the transverse true FISP sequence and 64%/86% (50%/82% for observer 2) for the coronal intermediate-weighted spin-echo images. The corresponding values for the glenoidal cartilage were 60% (6/10)/88% (51/58) (80%/76% for observer 2) and 70%/86% (60%/74% for observer 2) respectively. For the detection of abnormalities of the anterior labrum (only assessed on true FISP images) the values were 94% (15/16)/84% (36/43) (88%/79% for observer 2). The corresponding values for the posterior labrum were 67% (8/12)/77% (36/47) (observer 2: 25%/74%). The kappa values for the grading of the humeral and glenoidal cartilage lesions were 0.81 and 0.55 for true FISP images compared with 0.49 and 0.43 for intermediate-weighted fast spin-echo images. Kappa values for true FISP evaluation of the anterior and posterior part of the labrum were 0.81 and 0.70. CONCLUSION: Transverse 3D true FISP MR arthrography images are useful for the difficult diagnosis of glenohumeral cartilage lesions and suitable for detecting labral abnormalities. PMID- 20016889 TI - The added value of multislice SPECT/CT in patients with equivocal bony metastasis from carcinoma of the prostate. AB - PURPOSE: The purpose of this study was to investigate the additional value of single photon emission computed tomography/computed tomography (SPECT/CT) over whole-body planar bone scintigraphy and SPECT in prostate cancer patients in terms of diagnostic confidence, inter-reviewer agreement and the possible impact on the clinical management. METHODS: This was a retrospective review of 40 consecutive prostate cancer patients (mean age 71 years) who underwent (99m)Tc methylene diphosphonate (MDP) whole-body planar bone scintigraphy, SPECT and SPECT/CT between April 2006 and April 2008. The images were evaluated by two independent reviewers; inter-reviewer agreement was evaluated using a weighted kappa score. Each focus of abnormal increased tracer uptake was recorded using a 4-point diagnostic confidence scale. Institutional Review Board approval was obtained. RESULTS: Fifty lesions on planar bone scintigraphy in the 40 patients were evaluated. On reporting the planar study and SPECT scans, reviewers rated 61% of lesions as equivocal. On reporting the SPECT/CT scans only 8% of lesions were rated as equivocal, 24% were rated as malignant and 68% as benign. Weighted kappa scores for inter-reviewer agreement were 0.43 for bone scintigraphy, 0.56 for SPECT and 0.87 for SPECT/CT. All were significant at p < 0.0001. Follow-up imaging confirmed the SPECT/CT diagnoses in 14 patients. CONCLUSION: The addition of SPECT/CT resulted in a significant reduction of equivocal reports; a definitive diagnosis was given in the majority of the patients due to the improved diagnostic confidence compared to planar or SPECT imaging alone in prostate cancer patients with suspected bone metastases. PMID- 20016891 TI - Active inflammation in 18F-methylcholine PET/CT. PMID- 20016890 TI - Dose-response assessment of tariquidar and elacridar and regional quantification of P-glycoprotein inhibition at the rat blood-brain barrier using (R) [(11)C]verapamil PET. AB - PURPOSE: Overactivity of the multidrug efflux transporter P-glycoprotein (P-gp) at the blood-brain barrier (BBB) is believed to play an important role in resistance to central nervous system drug treatment. (R)-[(11)C]verapamil (VPM) PET can be used to measure the function of P-gp at the BBB, but low brain uptake of VPM hampers the mapping of regional differences in cerebral P-gp function and expression. The aim of this study was to evaluate the dose-response relationship of two potent P-gp inhibitors and to investigate if increased brain uptake of VPM mediated by P-gp inhibition can be used to assess regional differences in P-gp activity. METHODS: Two groups of Sprague-Dawley rats (n = 12) underwent single VPM PET scans at 120 min after administration of different doses of the P-gp inhibitors tariquidar and elacridar. In an additional six rats, paired VPM PET scans were performed before and after administration of 3 mg/kg tariquidar. RESULTS: Inhibitor administration resulted in an up to 11-fold increase in VPM brain distribution volumes (DV) with half-maximum effective dose (ED(50)) values of 3.0 +/- 0.2 and 1.2 +/- 0.1 mg/kg for tariquidar and elacridar, respectively. In paired PET scans, 3 mg/kg tariquidar resulted in regionally different enhancement of brain activity distribution, with lowest DV in cerebellum and highest DV in thalamus. CONCLUSION: Our data show that tariquidar and elacridar are able to increase VPM brain distribution in rat brain up to 11-fold over baseline at maximum effective doses, with elacridar being about three times more potent than tariquidar. Regional differences in tariquidar-induced modulation of VPM brain uptake point to regional differences in cerebral P-gp function and expression in rat brain. PMID- 20016892 TI - Efficacy of using a standard activity of (131)I-MIBG therapy in patients with disseminated neuroendocrine tumours. AB - PURPOSE: The aim of this analysis was to evaluate the response to standard activity of (131)I-meta-iodobenzylguanidine (MIBG) in patients with disseminated neuroendocrine tumours (NETs), comparing overall survival of patients with symptomatic response, tumour size (as assessed by CT) and relevant plasma tumour markers. METHODS: A retrospective review of patients who had undergone (131)I MIBG treatment between March 2001 and December 2006 was carried out. The administered activity of (131)I-MIBG was 5.5 GBq (NETs) and 7 GBq (phaeochromocytoma). Three cycles of treatment were planned with an interval of 10-12 weeks. A pre-therapy scan with (123)I-MIBG was performed to ascertain appropriate biodistribution. RESULTS: Thirty-eight patients were identified. Only two patients developed significant bone marrow suppression. Symptomatic response: data were available in 37 of 38 patients: 15 patients had improved symptoms, 19 had no improvement in symptoms and 3 were asymptomatic. In those with a symptomatic response, the median overall survival was 58 months vs no response of 20.0 months (p = 0.001). CT response: in those with stable disease, the median overall survival was 58 months compared with progressive disease of 16.0 months. The difference between these groups was significant (p = 0.006). Hormonal response: this was available in only 20 of 38 patients. The median overall survival was the same for patients that had increased hormone levels and patients that had stable/decreased hormone levels (48 months). CONCLUSION: Standard activity (131)I-MIBG is well tolerated. Symptomatic response to treatment is a significant predictor of overall survival. Whilst CT response also appears to predict survival, hormonal levels do not appear to correlate with survival. PMID- 20016893 TI - Pulmonary artery and femoral vein tumour thromboembolism in a patient with osteogenic sarcoma demonstrated by FDG PET/CT. PMID- 20016894 TI - Significance of cardiac sympathetic nervous system abnormality for predicting vascular events in patients with idiopathic paroxysmal atrial fibrillation. AB - PURPOSE: Neuronal system activity plays an important role for the prognosis of patients with atrial fibrillation (AF). Using (123)I metaiodobenzylguanidine ((123)I-MIBG) scintigraphy, we investigated whether a cardiac sympathetic nervous system (SNS) abnormality would be associated with an increased risk of vascular events in patients with paroxysmal AF. METHODS: (123)I-MIBG scintigraphy was performed in 69 consecutive patients (67 + or - 13 years, 62% men) with paroxysmal AF who did not have structural heart disease. SNS integrity was assessed from the heart to mediastinum (H/M) ratio on delayed imaging. Serum concentration of C-reactive protein (CRP) was measured before (123)I-MIBG study. RESULTS: During a mean of 4.5 + or - 3.6 years follow-up, 19 patients had myocardial infarction, stroke or heart failure (range: 0.2-11.5 years). SNS abnormality (H/M ratio <2.7) and high CRP (> or = 0.3 mg/dl) were associated with the vascular events (58.3% in 14 of 24 patients with SNS abnormality vs 11.1% in 5 of 45 patients without SNS abnormality, p < 0.0001, 52.4% in 11 of 21 patients with high CRP vs 16.7% in 8 of 48 patients without high CRP, p < 0.0001). After adjustment for potential confounding variables such as age, left atrial dimension and left ventricular function, SNS abnormality was an independent predictor of vascular events with a hazard ratio of 4.1 [95% confidence interval (CI): 1.3 12.6, p = 0.014]. Further, SNS abnormality had an incremental and additive prognostic power in combination with high CRP with an adjusted hazard ratio of 4.1 (95% CI: 1.5-10.9, p = 0.006). CONCLUSION: SNS abnormality is predictive of vascular events in patients with idiopathic paroxysmal AF. PMID- 20016895 TI - In vivo detection of prion amyloid plaques using [(11)C]BF-227 PET. AB - PURPOSE: In vivo detection of pathological prion protein (PrP) in the brain is potentially useful for the diagnosis of transmissible spongiform encephalopathies (TSEs). However, there are no non-invasive ante-mortem means for detection of pathological PrP deposition in the brain. The purpose of this study is to evaluate the amyloid imaging tracer BF-227 with positron emission tomography (PET) for the non-invasive detection of PrP amyloid in the brain. METHODS: The binding ability of BF-227 to PrP amyloid was investigated using autoradiography and fluorescence microscopy. Five patients with TSEs, including three patients with Gerstmann-Straussler-Scheinker disease (GSS) and two patients with sporadic Creutzfeldt-Jakob disease (CJD), underwent [(11)C]BF-227 PET scans. Results were compared with data from 10 normal controls and 17 patients with Alzheimer's disease (AD). The regional to pons standardized uptake value ratio was calculated as an index of BF-227 retention. RESULTS: Binding of BF-227 to PrP plaques was confirmed using brain samples from autopsy-confirmed GSS cases. In clinical PET study, significantly higher retention of BF-227 was detected in the cerebellum, thalamus and lateral temporal cortex of GSS patients compared to that in the corresponding tissues of normal controls. GSS patients also showed higher retention of BF-227 in the cerebellum, thalamus and medial temporal cortex compared to AD patients. In contrast, the two CJD patients showed no obvious retention of BF-227 in the brain. CONCLUSION: Although [(11)C]BF-227 is a non specific imaging marker of cerebral amyloidosis, it is useful for in vivo detection of PrP plaques in the human brain in GSS, based on the regional distribution of the tracer. PET amyloid imaging might provide a means for both early diagnosis and non-invasive disease monitoring of certain forms of TSEs. PMID- 20016896 TI - Long-term prognostic value of early poststress (99m)Tc-tetrofosmin lung uptake during exercise (SPECT) myocardial perfusion imaging. AB - PURPOSE: The aim of this study was to determine the long-term prognostic value of early poststress lung/heart ratio (LHR) of (99m)Tc-tetrofosmin radioactivity. METHODS: We studied 276 patients (aged 62.2 + or - 8.9 years, 168 men) with stress/rest (99m)Tc-tetrofosmin myocardial gated-SPECT and coronary angiography. To evaluate myocardial ischaemia, we calculated the summed stress score, summed rest score and summed difference score indices. For the eLHR calculation, an anterior image was acquired, 4-6 min after radiotracer injection at stress (eLHR was defined as mean counts per pixel in the lung region of interest divided by the mean counts per pixel in the myocardial region of interest). Cardiovascular death and nonfatal myocardial infarction were considered as hard cardiac events, and late revascularization procedures as soft cardiac events. The Cox proportional hazards model in a stepwise method was used to determine the independent predictors for hard and soft cardiac events. RESULTS: During the follow-up period hard cardiac events occurred in 28 patients (10.1%) and soft cardiac events in 32 patients (11.6%). Implying multiple Cox regression analysis, eLHR was found to be a significant independent predictor for both soft and hard cardiac events. The hazard ratio (for a 0.1 unit increase) was 4.41 (95% CI 1.52 12.73, p=0.006) for soft cardiac events and 4.22 (95% CI 2.07-8.62, p<0.001) for hard cardiac events. The other significant prognostic factors were use of beta blockers, the summed stress score and the use of nitrates for soft events, and exercise duration and the summed stress score for hard cardiac events. CONCLUSION: Early poststress (99m)Tc-tetrofosmin LHR has an independent and powerful value in predicting hard and soft cardiac events. PMID- 20016897 TI - Aortoesophageal fistula and aortic pseudoaneurysm induced by swallowed fish bone: a report of two cases. AB - Esophageal perforation caused by accidental swallowing of fish bones can lead to rare complications, such as aortoesophageal fistula accompanied by aortic pseudoaneurysm, which can be fatal if not properly handled. We report two rare cases of aortoesophageal fistula and aortic pseudoaneurysm caused by esophagus perforation after accidental swallow of fish bone; the patients also had purulent mediastinitis and esophagitis. The treatment of aortic pseudoaneurysm was successful in both cases, with one patient undergoing surgical resection and aortic neoplasty and the other patient undergoing endovascular stent graft placement. Long-term antibiotic treatment was administered to both patients after surgery. There were no postsurgical complications, and the patients recovered without incident. PMID- 20016898 TI - Absence of cardiac siderosis despite hepatic iron overload in Italian patients with thalassemia intermedia: an MRI T2* study. AB - Cardiac involvement in patients with thalassemia intermedia (TI) is characterized by a high-output state and pulmonary hypertension, with systolic left ventricle function usually being preserved. Myocardial iron overload in patients with TI has not been extensively studied. We conducted a cross-sectional study of 49 Italian patients with TI. Patient charts were reviewed and data collected for transfusion and iron chelation history, status of the spleen, and comorbid illnesses or infections. Blood samples were obtained for assessment of hemoglobin, serum ferritin, and liver enzyme levels. Doppler echocardiography was done for all patients. Cardiac and hepatic iron levels were measured by magnetic resonance imaging T2*. The mean age was 40.5 +/- 8.3 years, with a male to female ratio of 29:20. A total of 34 (69.4%) patients were splenectomized, and four patients had evidence of hepatitis C infection. Around 45% of patients were transfusion naive while the rest received infrequent (47%) or regular (8%) transfusions. A total of 31 (63.3%) patients were maintained on iron chelation therapy. None of the patients had evidence of heart failure. Mean serum ferritin and liver iron concentration were 1,060.2 ng/ml and 8.2 mg Fe per gram dry weight, respectively. None of the patients had evidence of cardiac iron overload (mean cardiac T2* = 38.7 +/- 11.0 ms). There were no statistically significant correlation between cardiac T2* values and liver iron concentration, serum ferritin, or any patient, disease, or treatment-related parameters. Patients with TI show absence of cardiac iron overload even if hepatic iron accumulation is significant. PMID- 20016899 TI - [Accreditation in pathology. Systematic presentation and documentation of activities in pathology]. AB - In the last ten years, almost 60 departments of surgical pathology were accredited in Germany according to DIN EN ISO/IEC 17020. Accreditation in pathology was accompanied by the adoption of a highly dynamic process, which requires staff to be more aware of quality and to introduce a quality-oriented system. The accreditation body in turn needed to consider all the duties, responsibilities and processes in surgical pathology and was supported in this regard by the Sector Committee of Pathology and Neuropathology of the DGA German Association for Accreditation. In this review we illustrate the various problems associated with accreditation in surgical pathology by answering eight questions that have arisen over many years of activity. These include the registration and appropriate presentation of the scope of accreditation, the terminology, as well as the extent and depth of documentation. A department of pathology applying for accreditation is required to document the entire step-wise process leading to a surgical pathological diagnosis. PMID- 20016900 TI - Feasibility of contrast material volume reduction in coronary artery imaging using 320-slice volume CT. AB - OBJECTIVE: To assess reduced volumes of contrast agent on image quality for coronary computed tomography angiography (CCTA) by using single-beat cardiac imaging with 320-slice CT. MATERIALS AND METHODS: Forty consecutive male patients (mean age: 55.8 years) undergoing CCTA with body weight or=55% were included. Image acquisition protocol was standardized (120 kV, 400 mA, and prospective ECG-triggered single beat nonspiral CCTA). Patients were randomly assigned to one of four groups (G1: received 40 ml, G2: 50 ml, G3: 60 ml, G4: 70 ml). Groups were compared with respect to aortic attenuation, image noise, and image quality. RESULTS: CT values (mean +/- standard deviation) in the aortic root were measured as 423 +/- 38 HU in G1, and 471 +/- 68, 463 +/- 60, and 476 +/- 78 HU in G2-4, respectively. There were no statistically significant differences in attenuation among the groups (P > 0.068). All 40 CT datasets were rated diagnostic, and image noise and image quality were not statistically different among groups. CONCLUSION: Using 320 slice volume CT, diagnostic image quality can be achieved with 40 ml of contrast material in CCTA in patients with normal body weight, cardiac function, and low heart rate. PMID- 20016901 TI - Thoracic causes of acute abdominal pain. AB - The origin of abdominal pain may be extra-abdominal, caused by a thoracic illness. This article illustrates the various thoracic disorders that may present with acute abdominal pain. An erroneous focus on the abdomen alone can easily lead to misdiagnosis and incorrect treatment. In cases of unexplained acute abdominal pain, radiologists should be aware of also viewing beyond the borders of the abdomen. The key to most of these thoracic diagnoses is detection of pulmonary consolidation, pleural fluid or pericardial fluid. PMID- 20016902 TI - Image-based clinical decision support for transrectal ultrasound in the diagnosis of prostate cancer: comparison of multiple logistic regression, artificial neural network, and support vector machine. AB - PURPOSE: We developed a multiple logistic regression model, an artificial neural network (ANN), and a support vector machine (SVM) model to predict the outcome of a prostate biopsy, and compared the accuracies of each model. METHOD: One thousand and seventy-seven consecutive patients who had undergone transrectal ultrasound (TRUS)-guided prostate biopsy were enrolled in the study. Clinical decision models were constructed from the input data of age, digital rectal examination findings, prostate-specific antigen (PSA), PSA density (PSAD), PSAD in transitional zone, and TRUS findings. The patients were divided into the training and test groups in a randomized fashion. Areas under the receiver operating characteristic (ROC) curve (AUC, Az) were calculated to summarize the overall performance of each decision model for the task of prostate cancer prediction. RESULTS: The Az values of the ROC curves for the use of multiple logistic regression analysis, ANN, and the SVM were 0.768, 0.778, and 0.847, respectively. Pairwise comparison of the ROC curves determined that the performance of the SVM was superior to that of the ANN or the multiple logistic regression model. CONCLUSION: Image-based clinical decision support models allow patients to be informed of the actual probability of having a prostate cancer. PMID- 20016904 TI - US-guided transcutaneous tru-cut biopsy of laryngo-hypopharyngeal lesions. AB - OBJECTIVE: To evaluate the feasibility and performance of ultrasound-guided transcutaneous tru-cut biopsy (USGTCB) in selected patients (with stenosis of airways or difficult intubation or contraindication to general anaesthesia) with untreated or previously treated suspicious laryngo-hypopharyngeal masses. METHODS: Biopsies were performed with a free-hand technique by a single radiologist. Thirty-six USGTCBs were scheduled in 34 patients (24 males, 10 females; age range 47-95 years). Two USGTCBs were not performed, as lesions were not detectable: therefore, 16 USGTCBs were performed for an untreated mass suspicious for malignancy and 18 were performed for a mass suspicious for recurrence after radiotherapy alone, or associated with endoscopic laser surgery or chemotherapy. RESULTS: USGTCB diagnosed 25 squamous cell carcinomas (73.5%) and nine benign lesions (26.5%); no false positives and two false negatives were reported, both in patients previously treated with radiotherapy. The sensitivity, specificity, positive and negative predictive value of the technique was 92.5%, 100%, 100% and 77.7% respectively, with no major complications. CONCLUSION: Although biopsy under microlaryngoscopy remains the "gold-standard" technique, USGTCB is feasible, carries the advantages of avoiding general anaesthesia, is suitable for outpatients and is cost-effective. If applied to selected patients, it could be considered for the histological diagnosis of both primary and recurrent laryngo-hypopharyngeal masses. PMID- 20016903 TI - Automatic postprocessing for the assessment of quantitative human myocardial perfusion using MRI. AB - OBJECTIVE: Quantitative determination of myocardial perfusion currently involves time-consuming postprocessing. This retrospective study presents automatic postprocessing consisting of image registration and image segmentation to obtain regional signal intensity time courses and quantitative perfusion values. METHODS: The automatic postprocessing was tested in 75 examinations in volunteers and patients, 57 at rest and 18 under adenosine-induced stress, and compared with a manual evaluation. In a substudy consisting of 10 examinations, the interobserver variability of the manual evaluation was investigated. RESULTS: Manual evaluation resulted in perfusion values with a median of 0.70 ml/g/min ranging from 0.03 to 3.68 ml/g/min. For all 75 examinations, the variability (standard deviation of the differences) between automatic and manual evaluation was 0.34 ml/g/min. Interobserver variability was of a similar order, 0.35 ml/g/min for all measurements. CONCLUSIONS: Automatic evaluation was successfully applied to all datasets giving results equivalent to manual evaluation. The time of user interaction for one single slice could be reduced from 25 min for manual evaluation to less than 1 min using the automatic algorithm. This reduction may allow quantitative magnetic resonance perfusion imaging to become a routine clinical procedure. PMID- 20016905 TI - Multidetector computed tomography hepatic findings in children with end-stage biliary atresia. AB - OBJECTIVES: To illustrate the multidetector computed tomography (MDCT) findings in patients with end-stage biliary atresia (BA). METHODS: The study group consists of 45 consecutive patients with BA who underwent MDCT before liver transplantation from February 2005 to February 2008. Mean age was 36 months, 24 patients were female, and 22 had undergone a previous Kasai procedure. RESULTS: MDCT detected a total of 15 hepatocellular nodules in 7 patients. Intrahepatic biliary cysts were detected in 14 patients and were significantly associated with a Kasai procedure. Intrahepatic porto-systemic shunts were found in 14 patients (31%), intrahepatic communicating vessels between hepatic veins were found in 24 patients (53%). Anatomical variants of hepatic artery were detected in 21 patients. Seven patients (15%) had portal vein thrombosis; in 12 cases (26%) portal vein diameter was 3 mm or less. CONCLUSION: MDCT can provide accurate morphological and vascular assessment of patients with end-stage biliary atresia and should be used for precise surgical planning. To the best of our knowledge this is one of the first studies to show the presence of numerous veno-venous communications in BA. PMID- 20016906 TI - Assessment of vascular invasion by bone and soft tissue tumours of the limbs: usefulness of MDCT angiography. AB - OBJECTIVE: To evaluate the accuracy of computed tomography angiography (CTA) in predicting arterial encasement by limb tumours, by comparing CTA with surgical findings (gold standard). METHODS: Preoperative CTA images of 55 arteries in 48 patients were assessed for arterial status: cross-sectional CTA images were scored as showing a fat plane between artery and tumour (score 0), slight contact between artery and tumour (score 1), partial arterial encasement (score 2) or total arterial encasement (score 3). Reformatted CTA images were assessed for arterial displacement, rigid wall, stenosis or occlusion. At surgery, arteries were classified as free or surgically encased; 45 arteries were free and 10 were surgically encased. RESULTS: Multivariate logistic regression identified the axial CTA score as a relevant predictor for arterial encasement and subsequent vascular intervention during surgery. All sites where CTA showed a fat plane between the tumour and the artery were classified as free at surgery (n = 28/28). The sensitivity of total arterial encasement on CTA (score 3) was 90%, specificity 93%, accuracy 93% and positive likelihood ratio 13.5. CONCLUSION: CTA evidence of total arterial encasement is a highly specific indication of arterial encasement. The presence of fat between the tumour and the artery on CTA rules out arterial involvement at surgery. PMID- 20016907 TI - [Deficits of routine occupational therapy services in rheumatic diseases: results of the RheumaDat study]. AB - The purpose of the present study is to describe the frequency and potential determinants of occupational therapy routine services in patients with rheumatic diseases. The study is based on a secondary analysis of the RheumaDat database. The results of the survey on 907 patients were analyzed regarding frequency of occupational therapy routine services and related sociodemographic and disease specific variables. Approximately 8% of the study population received occupational therapy, patients with rheumatoid arthritis representing the largest proportion of these at 13%. Only patients with osteoarthritis showed a clear treatment pattern. Patients from this group receiving occupational therapy were older, had a longer duration of disease and were more affected. No clear appraisal of the therapy status in the rheumatoid arthritis or fibromyalgia group could be made. The results indicate shortages and the lack of a clear system in occupational therapy routine services in patients with different rheumatic diseases. PMID- 20016908 TI - Altered sleep architecture and higher incidence of subsyndromal depression in low endogenous melatonin secretors. AB - Melatonin secretion is synchronized to the sleep/wake cycle and has been suggested to have somnogenic properties. Sleep/wake cycle disruption and alterations in the secretary pattern of melatonin is present in various psychiatric disorders. The objective of this study was to investigate the sleep architecture and the presence of depression in individuals with low endogenous melatonin levels. The study included 16 participants (mean age 30.3 +/- 14.9 years). The first night of testing included psychiatric evaluation followed by melatonin secretion profile evaluation by Dim Light Melatonin Onset test and then standard montage polysomnographic testing. On the second night, only polysomnographic testing was carried out with an imposed sleep period of 8 h. Low endogenous melatonin secretors (LEMS) showed no discernible peaks in melatonin secretion compared to normal secretors (controls). LEMS demonstrated significant alterations in rapid eye movement sleep but not in non-rapid eye movement sleep along with poor sleep initiation and quality compared to controls. 55.6% of the low melatonin secretors group presented with subsyndromal depression. Melatonin has significant bearing on sleep architecture and a lack of melatonin may desynchronize endogenous rhythms allowing subsyndromal depression to manifest. PMID- 20016909 TI - B chromosome ancestry revealed by histone genes in the migratory locust. AB - In addition to the standard set of chromosomes (A), about 15% of eukaryote genomes carry B chromosomes. In most cases, B chromosomes behave as genomic parasites being detrimental for the individuals carrying them and prospering in natural populations because of transmission advantages (drive). B chromosomes are mostly made up of repetitive DNA sequences, especially ribosomal DNA (rDNA), satellite DNA and mobile elements. In only two cases have B chromosomes been shown to carry protein-coding genes. Although some B chromosomes seem to have derived from interspecific hybridisation, the most likely source of B chromosomes is the host genome itself, but the specific A chromosome being the B ancestor has not been identified in any B-containing species. Here, we provide strong evidence for B chromosome ancestry in the migratory locust, based on the location of genes for the H3 and H4 histones in the B chromosome and a single A chromosome pair (i.e. the eighth in order of decreasing size). The high DNA sequence similarity of A and B chromosome H3-H4 genes supports B-origin from chromosome 8. The higher variation shown by B sequences, compared to A sequences, suggests that B chromosome sequences are most likely inactive and thus less subjected to purifying selection. Estimates of time of divergence for histone genes from A and B chromosomes suggest that B chromosomes are quite old (>750,000 years), showing the B-chromosome ability to persist in natural populations for long periods of time. PMID- 20016910 TI - Changing causes of enucleation over the past 60 years. AB - BACKGROUND: The indications for enucleation have changed significantly over the past 60 years. We conducted a clinicopathologic study of enucleated globes to determine how and why the indications for enucleation have changed over time. METHODS: This retrospective review examined the pathology reports for 3,264 enucleated globes submitted to the Doheny Eye Institute between 1950 and 2006. Three years per decade were examined to generate a representative pool of specimens for each decade. Although the data for the 2000s were only available up to 2006, the data for this decade are drawn from 3 sample years as are all other decades. Pathology reports were reviewed for demographic information (age, sex, and ethnicity), clinical history prior to enucleation, and pathologic findings and diagnoses. Specimens were grouped according to the reason for enucleation into the following categories: atrophic/phthisis bulbi, congenital, glaucoma, infection, longstanding retinal detachment, trauma, tumor, uveitis, and other. RESULTS: During the study period, there were 3,264 enucleated globes. Overall, the total number of enucleations decreased over time from a peak of 1,014 in the 1960s to 275 in the 2000s. Glaucoma was the most common reason for enucleation during the 1950s (23%, 127 globes) and 1960s (31%, 315 globes). However, glaucoma steadily decreased over the following decades, and was responsible for only 8% (23 globes) of enucleations in the 2000s. Neovascular glaucoma (including glaucoma secondary to retinal vein occlusion and diabetic neovascularization) accounted for 21% (27 globes) of enucleations in the 1950s. By the 2000s, this number was 57% (13 globes). Trauma-related glaucoma accounted for 34% (43 globes) of all enucleations due to glaucoma in the 1950s, and 0% (0 globes) in the 2000s. Enucleation of globes with intraocular neoplasms accounted for 14% (79 globes) of total enucleations in the 1950s, 33% (120 globes) in the 1990s, and 51% (141 globes) in the 2000s. Uveal melanoma was the main intraocular neoplasm in the 1950s (77%, 60 globes), and retinoblastoma was the primary tumor in the enucleated globes of the 2000s (69%, 97 globes). CONCLUSIONS: Improved medical and surgical treatment of conditions that lead to end-stage eye disease have led to a decrease in total enucleated globes. This is particularly evident for glaucoma. Changing demographics in Los Angeles and referral patterns are most likely responsible for the increase in retinoblastoma. The absolute number of enucleations secondary to neoplasms has not decreased over time, despite an increase in globe-conserving treatments such as chemotherapy and radioactive plaques. PMID- 20016911 TI - The influence of gestational age on the dynamic behavior of other risk factors associated with retinopathy of prematurity (ROP). AB - BACKGROUND: Improved survival of preterm neonates has increased the incidence of retinopathy of prematurity (ROP) in many middle-income countries. AIM: This study aimed to verify the main risk factors for the development of ROP according to different gestational age (GA) groups. METHODS: A prospective cohort study including infants weighing < or =1,500 g or GA < or =32 weeks at birth was conducted. The main clinical outcomes were the occurrence of any stage of ROP and severe ROP. The perinatal variables considered for the study were: birth weight; GA; gender; to be small for GA (SGA); weight gain from birth to the sixth week of life; use of oxygen in mechanical ventilation or nasal CPAP; multiple gestations; therapeutic use of surfactant, indomethacin, and erythropoietin; occurrence of sepsis, meningitis, intraventricular hemorrhage, and patent ductus arteriosus; need for and volume of blood transfusion; and 10-min Apgar score. The patients were divided into three groups according to GA: (group 1) infants of GA < or =28 weeks at birth (n = 100); (group 2) infants of GA = 29-31 weeks at birth (n = 215); and (group 3) infants of GA > or =32 weeks at birth (n = 152). RESULTS: A total of 467 newborn infants were included. Mean BW and GA in the total cohort were 1,216.5 g (+/-278.3) and 30.3 weeks (+/-2.2), respectively. Gestational age groups were not matched for BW and SGA. Any stage of ROP occurred in 111 patients (23.8%) and 24 (5.1%) patients developed severe ROP. Only BW and volume of blood transfusion were significant factors for the occurrence of any stage of ROP in all groups. In group 1, GA, the twin situation, and use of erythropoietin were statistically significant factors. In group 2, only GA and need for blood transfusion were significant. In group 3, use of oxygen in mechanical ventilation, sepsis, and need for blood transfusion were significant for ROP onset. The logistic regression determined that patients in groups 2 and 3 were less likely to develop ROP than patients in group 1. CONCLUSIONS: Patients in groups 1 and 2 developed ROP due to general immaturity, whereas bigger babies, of GA > or =32 weeks, developed ROP because they were "sicker" babies with more comorbidities. PMID- 20016912 TI - Conserved intron positions in FGFR genes reflect the modular structure of FGFR and reveal stepwise addition of domains to an already complex ancestral FGFR. AB - We have analyzed the evolution of fibroblast growth factor receptor (FGFR) tyrosine kinase genes throughout a wide range of animal phyla. No evidence for an FGFR gene was found in Porifera, but we tentatively identified an FGFR gene in the placozoan Trichoplax adhaerens. The gene encodes a protein with three immunoglobulin-like domains, a single-pass transmembrane, and a split tyrosine kinase domain. By superimposing intron positions of 20 FGFR genes from Placozoa, Cnidaria, Protostomia, and Deuterostomia over the respective protein domain structure, we identified ten ancestral introns and three conserved intron groups. Our analysis shows (1) that the position of ancestral introns correlates to the modular structure of FGFRs, (2) that the acidic domain very likely evolved in the last common ancestor of triploblasts, (3) that splicing of IgIII was enabled by a triploblast-specific insertion, and (4) that IgI is subject to substantial loss or duplication particularly in quickly evolving genomes. Moreover, intron positions in the catalytic domain of FGFRs map to the borders of protein subdomains highly conserved in other serine/threonine kinases. Nevertheless, these introns were introduced in metazoan receptor tyrosine kinases exclusively. Our data support the view that protein evolution dating back to the Cambrian explosion took place in such a short time window that only subtle changes in the domain structure are detectable in extant representatives of animal phyla. We propose that the first multidomain FGFR originated in the last common ancestor of Placozoa, Cnidaria, and Bilateria. Additional domains were introduced mainly in the ancestor of triploblasts and in the Ecdysozoa. PMID- 20016913 TI - The epidemiology of childhood poisonings in Cyprus. AB - Information on childhood poisonings in Cyprus is limited. Our objective was to examine the epidemiology of poisonings among children in Cyprus. All children up to 15 years of age admitted for poisoning to the Archbishop Makarios Hospital in Nicosia, Cyprus between 2005 and 2008 were included in our study. All hospital poisoning records were reviewed. A total of 257 children were admitted for poisoning. The mean age of children was 3.1 years, of which 83.7% were below the age of 5 years old, while 53% were boys. The poisoning hospitalizations accounted for about 3% of all admissions to the pediatric department during the study period (4 years). The annual cumulative incidence of childhood poisoning hospitalizations was 116 per 100,000 children. Medications accounted for 46.1% of all poisonings, the most frequent cause being paracetamol (9.8%), cardiovascular medications (5.3%), antitussive medications (4.5%), and other painkillers (4.1%). Another 37.6% of hospitalizations involved household products such as household cleaning products (11.8%), petroleum products (11.0%), and rodenticides (5.7%). Among children who ingested petroleum distillates, 55.6% developed clinical symptomatology. The vast majority of cases were accidental (93.8%). Suicidal cases involved children 8-14 years old, mainly girls, and the most frequent poisoning ingested was paracetamol (46.7%). Poisoning hospitalizations represent an important cause of morbidity among children in Cyprus. Preventive strategies should include the education of caregivers on the handling of medications and household products as well as legislation requiring child-resistant packaging for all medications and household products including petroleum distillates. PMID- 20016914 TI - Severe hypernatremia in newborns due to salting. AB - The aim of this study was to evaluate the etiology, clinical, and laboratory findings and prognostic features of life-threatening hypernatremic newborns secondary to salting. Ten severely hypernatremic newborns (four females) with a mean age of 6.5 +/- 2.6 days were followed up. Nine of them were full term, and one was preterm. It was noticeable that 60% of them were small for gestational age. In the laboratory investigation, five uremias were detected. It was interesting to find in the etiologic history that 40% of the patients had been salted just after birth. Twenty percent of them had also hyperbilirubinemia and kernicterus, 20% had neonatal convulsion, and 50% had dehydration. Two of the hypernatremic newborns died during the study; the others were followed up. One case had spasticity and developmental disability at the 3rd month, and another one had developmental disability at the 6th month of ages. As a conclusion, although salting of newborns is not so frequent, it could be seen in rural places of our country, and this may be one of the reasons for serious hypernatremia in newborns whose skin integrity have not been formed completely. These cases should be treated carefully. PMID- 20016915 TI - Purpura fulminans in a newborn infant with galactosemia. AB - An 11-day-old neonate presented with purpura fulminans and was subsequently diagnosed with galactosemia. Neonatal purpura fulminans occurs predominantly in patients suffering from inherited protein C deficiency or disseminated intravascular coagulation associated with septicemia. Hemostatic changes in patients with liver disease may result in bleeding or, rarely, thrombosis. We suppose that in the present patient, deficiency of protein C, secondary to liver disease, was responsible for the development of purpura fulminans. Treatment consisted of blood and blood products and galactose-free formula. The patient recovered with residual mild psychomotor retardation and the lesions with minimal scarring. In conclusion, galactosemia also should be kept in mind as an uncommon cause of purpura fulminans in newborn infants. PMID- 20016916 TI - The effect of colchicine on physical growth in children with familial mediterranean fever. AB - Familial Mediterranean fever (FMF) is an autosomal recessive disease, characterized by recurrent, self-limited attacks of fever with serositis involving the peritoneum, pleura, and joints. There is very scarce information on physical growth of affected children. The aim of this study was to determine whether there is significant improvement in growth parameters in FMF patients after colchicine treatment. Patient files were retrospectively evaluated and patients that used colchicine for more than 1 year were included in the study. Demographic features, clinical findings before and after colchicine therapy, duration and dosage of therapy, weight, height, parentally adjusted height, and body mass index before and after colchicine therapy were noted and transformed into standard deviation scores (SDS). The study group consisted of 50 FMF (25 male and 25 female) patients. Median age at the time of diagnosis was 6.5 years. Median follow-up period was 3.6 (1-12.5) years. Mean height SDS increased from 0.19 +/- 1.01 to 0.13 +/- 0.99 (p = 0.026), and mean parentally adjusted height increased from -0.18 +/- 1.23 to 0.13 +/- 1.24 (p = 0.027), and both of them were found to be statistically significant. Mean body mass index SDS increased from 0.61 +/- 1.32 to -0.32 +/- 1.33, but this improvement was statistically insignificant (p = 0.18). In this study, we found that colchicine significantly improved height development in FMF patients. PMID- 20016917 TI - The Michel Benoist and Robert Mulholland yearly European Spine Journal Review: a survey of the "medical" articles in the European Spine Journal, 2009. PMID- 20016918 TI - Continuous thrombolysis and repeated thrombectomy with the Penumbra System in a child with hemorrhagic sinus thrombosis: technical note. AB - Cerebral venous sinus thrombosis (CVST) is an uncommon but potentially lethal event. When showing a malignant clinical course despite anticoagulant therapy, new endovascular techniques and technology allow the possibility of more aggressive thrombolysis and thrombectomy. The authors present a case of recanalization of an extensive cerebral thrombosis using a new endovascular thromboaspiration device, the Penumbra System. BACKGROUND AND PURPOSE: CVST in children is a rare but potentially lethal disorder. When showing a malignant progression despite anticoagulant therapy, the use of local thrombolytics and thrombectomy procedures might provide therapeutic solution. METHODS: A 16-year old girl presented with signs and symptoms of a hemorrhagic cerebral sinus thrombosis and showed progression despite full dose anticoagulant therapy. The evolution being attributed to increasing mass effect, decompressive surgery was performed without clinical improvement. Further brain imaging demonstrated progression of the sinus thrombosis. Repeated local thrombectomy with the Penumbra thrombectomy system and continuous, relatively low dose local thrombolysis was performed during a period of 60 h, resulting in recanalization and clinical improvement. The methodology of combined thromboaspiration and thrombolysis is described. CONCLUSIONS: We present a novel method for revascularization using the Penumbra System in combination with long-term, relatively low dose thrombolysis, in the setting of both clinically and morphologically progressive, hemorrhagic sinus thrombosis. The procedure proved to be both safe and effective and may be considered in the future for patients with progressive sinovenous thrombosis despite adequate medical therapy. PMID- 20016919 TI - Genetic diversity of the S10 RNA segment of field and vaccine strains of bluetongue virus from the P. R. China. AB - Bluetongue virus (BTV) infection of ruminants is endemic throughout tropical and subtropical regions of the world. However, the molecular epidemiology of BTV infection in China has not yet been reported. In this study, the S10 gene segments from 30 BTV isolates, one attenuated BTV strain, one vaccine BTV strain, and one South Africa BTV prototype strain, were sequenced. Phylogenetic analysis of the S10 genes showed that Chinese BTV isolates could be classified into two phyletic subgroups, and the clustering of Chinese BTV viruses was dependent on their geographical origin and the number of generations for which they had been propagated, rather than their host species or year of isolation. PMID- 20016920 TI - Structure and genome organization of the large RNA of iris yellow spot virus (genus Tospovirus, family Bunyaviridae). AB - The structure and organization of the large (L) RNA of iris yellow spot virus (IYSV) was determined, and with this report, the complete genomic sequence of IYSV of the genus Tospovirus, family Bunyaviridae has been elucidated. The L RNA of IYSV was 8,880 nucleotides in length and contained a single open reading frame in the viral complementary (vc) strand. The primary translation product of 331.17 kDa shared many of the features of the viral RNA-dependent RNA polymerase (RdRp) coded by L RNAs of known tospoviruses. The 5' and 3' termini of IYSV L RNA (vc) contain two untranslated regions of 33 and 226 nucleotides, respectively, and both termini have conserved terminal nucleotides, another common feature of tospovirus genomic RNAs. Conserved motifs characteristic of RdRps of members of the family Bunyaviridae were present in the IYSV RdRp. PMID- 20016921 TI - Identification of cellular factors binding to acetylated HIV-1 integrase. AB - The viral protein integrase (IN) catalyzes the integration of the HIV-1 cDNA into the host cellular genome. We have recently demonstrated that IN is acetylated by a cellular histone acetyltransferase, p300, which modifies three lysines located in the C-terminus of the viral factor (Cereseto et al. in EMBO J 24:3070-3081, 2005). This modification enhances IN catalytic activity, as demonstrated by in vitro assays. Consistently, mutations introduced in the targeted lysines greatly decrease the efficiency of HIV-1 integration. Acetylation was proven to regulate protein functions by modulating protein-protein interactions. HIV-1 to efficiently complete its replication steps, including the integration reaction, requires interacting with numerous cellular factors. Therefore, we sought to investigate whether acetylation might modulate the interaction between IN and the cellular factors. To this aim we performed a yeast two-hybrid screening that differs from the screenings so far performed (Rain et al. in Methods 47:291-297, 2009; Studamire and Goff in Retrovirology 5:48, 2008) for using as bait IN constitutively acetylated. From this analysis we have identified thirteen cellular factors involved in transcription, chromatin remodeling, nuclear transport, RNA binding, protein synthesis regulation and microtubule organization. To validate these interactions, binding assays were performed showing that acetylation increases the affinity of IN with specific factors. Nevertheless, few two-hybrid hits bind with the same affinity the acetylated and the unmodified IN. These results further underlie the relevance of IN post translational modification by acetylation in HIV-1 replication cycle. PMID- 20016922 TI - Natural history of Charcot-Marie-Tooth 2: 2-year follow-up of muscle strength, walking ability and quality of life. AB - Charcot-Marie-Tooth (CMT) disease is the most frequent inherited neuropathy, no therapies are available at the moment but clinical trials are ongoing. For that reason it is very important to know the natural history of the disease. We report the results of the natural history of clinical features and quality of life (QoL) in patients with CMT2. Twenty patients were enrolled. At recruitment and at follow-up (2 years), all patients underwent neurological evaluation, QoL and disability assessments. The study-end evaluation took place 20-28 months after the baseline evaluation. During the 2-year follow-up period, CMT2 patients showed a mild reduction of strength of distal muscles of upper limbs and proximal muscles of lower limbs, a worsening sensory function and a mild increase in walking disability. However, there was no relevant worsening of QoL, except for a mild deterioration of one mental health domain. PMID- 20016923 TI - Influence of different power outputs of erbium, chromium:yttrium-scandium-gallium garnet laser and acid etching on shear bond strengths of a dual-cure resin cement to enamel. AB - The purpose of the study was to determine if the irradiation of enamel with laser of different output powers might be viable alternatives to acid etching for the bonding of resin luting agents. Seventy-seven maxillary central incisors, extracted for periodontal reasons, were used. The enamel was etched with an erbium, chromium:yttrium-scandium-gallium-garnet (Er,Cr:YSGG) laser operated at one of six power outputs (0.5 W, 0.75 W, 1 W, 1.5 W, 1.75 W and 2 W) or with 38% phosphoric acid. Seventy teeth were used for the bond strength experiments, and the remaining seven (one specimen for each group) were used for scanning electron microscopy (SEM) to determine the topography and morphology of the treated enamel surface. The acid-etched group yielded the highest mean of shear bond strength (13.5 +/- 2.8 MPa). The means of the shear bond strength for the teeth irradiated at 0.5 W, 0.75 W, 1 W, 1.5 W , 1.75 W and 2 W laser were 3.28 +/- 2.4 MPa, 5.44 +/- 3.4 MPa, 8.8 +/- 4.5 MPa, 10.2 +/- 4.0 MPa, 11.4 +/- 4.8 MPa and 11.9 +/- 4.3 MPa, respectively. Laser irradiation at 1.5 W, 1.75 W and 2 W produced a type III acid-etched pattern similar to that produced by acid etching. No significant enamel surface etching was obtained by 0.5 W or 0.75 W laser irradiation. Irradiation at 0.5 W and 0.75 W produced a type V acid-etched pattern. We concluded that the mean shear bond strength and enamel surface etching obtained with Er,Cr:YSGG laser (operated at 1.5 W and 1.75 W for 15 s) is comparable to that obtained with acid etching. PMID- 20016924 TI - Renal involvement in leprosy: report of progression from diffuse proliferative to crescentic glomerulonephritis. AB - Renal involvement in leprosy has been reported rarely in the literature. Acute kidney injury in patients with leprosy is uncommon and may occur due to acute tubular necrosis, drug-induced interstitial nephritis and rarely crescentic glomerulonephritis. The latter with histologic confirmation of the diagnosis has been reported in very few cases of leprosy. A 25-year-old male, on therapy for multibacillary leprosy, was found to have deranged renal functions on evaluation for a history of nausea, vomiting, swelling and episode of haematuria. Kidney biopsy was performed twice over a period of 2 weeks, showing progression from diffuse proliferative glomerulonephritis to crescentic glomerulonephritis, pauci immune in nature. The patient was treated aggressively with intravenous steroids, following which his renal functions stabilized. Crescentic glomerulonephritis, an extremely rare phenomenon in leprosy, should be considered in these patients presenting with features of acute kidney injury. Timely performed renal biopsy assists in accurate diagnosis and appropriate management of the patient, hence preserving renal parenchyma. Rapid progression from diffuse proliferative glomerulonephritis to crescentic glomerulonephritis in a patient with leprosy is described herein for the first time in the literature. PMID- 20016925 TI - Femoral fracture following knee ligament reconstruction surgery due to an unpredictable complication of bioabsorbable screw fixation: a case report and review of literature. AB - We report an unusual case of femoral fracture from minimal trauma, due to the rapid disappearance of a bioabsorbable interference screw used for reconstruction of the posterolateral corner of the knee. The literature on bone tunnel fractures following knee ligament reconstruction surgery is also reviewed. PMID- 20016926 TI - Deficiencies in elements involved in TLR4-receptor signalling in RBL-2H3 cells. AB - OBJECTIVE: RBL-2H3 cells express Toll-like receptors, including TLR4. This study aims to assess various aspects of the TLR4 pathway. METHODS: RBL-2H3 cells were indirectly stained for cell surface TLR4, 25 CD14 and intracellular MyD88 proteins and analysed through flow cytometry for single-colour staining. RESULTS: While TLR4-receptors are expressed in RBL-2H3 cells, associated elements involved in the signaling pathway, CD14 and MyD88, are not. CONCLUSION: Care should be taken if RBL-2H3 cells are used to study aspects of the innate immune system in mast cells. PMID- 20016927 TI - Efficacy and safety of sorafenib in a subset of patients with advanced soft tissue sarcoma from a Phase II randomized discontinuation trial. AB - AIM: Phase II multi-disease randomized discontinuation trial to assess the safety and efficacy of sorafenib including patients with advanced soft tissue sarcoma (STS). METHODS: Sorafenib (400 mg twice daily) was initially administered for 12 weeks. Patients with: >=25% tumour shrinkage continued sorafenib; >=25% tumour growth discontinued; other patients were randomized and received sorafenib or placebo. RESULTS: Twenty-six patients (median age 55 years) were enrolled. Common drug-related adverse events, including fatigue, hand-foot skin reaction, rash or gastrointestinal disturbances, were manageable, reversible and generally low grade. Fatigue, skin toxicity, nausea, diarrhoea and hypertension occurred at grade >=3 in 19% of patients. After 12 weeks eight (31%) patients had not progressed. Three patients who experienced tumour shrinkage and continued on sorafenib, and five (19%) were randomized either to continue sorafenib or to receive placebo. Of the three patients randomized to sorafenib, one achieved a partial response and two had SD. Overall one patient achieved a partial response and three further patients achieved minor responses. CONCLUSIONS: There was evidence of disease activity in STS as defined by tumor regressions including one objective partial response. Further investigation in STS is warranted. PMID- 20016928 TI - Evaluation of cell viability, DNA damage, and cell death in normal human dermal fibroblast cells induced by functionalized multiwalled carbon nanotube. AB - Multiwalled carbon nanotubes (MWCNTs) are an example of a carbon-based nanomaterial that has won enormous popularity in nanotechnology. Due to their unusual one-dimensional hollow nanostructure and unique physicochemical properties, they are highly desirable for use within the commercial, environmental, and medical sectors. Despite their wide application, there is a lack of information concerning their impact on human health and the environment. While nanotechnology looms large with commercial promise and potential benefit, an equally large issue is the evaluation of potential effects on humans and other biological systems. Our research is focused on cellular response to purified functionalized MWCNT in normal human dermal fibroblast cells. Three exposure concentrations (40, 200, and 400 microg/ml) of functionalized MWCNT and control (Tween-80 + 0.9% saline) were used in this study. Following exposure to MWCNT, cytotoxicity, genotoxicity, and apoptosis assays were performed using standard protocols. Our results demonstrated a dose-dependent toxicity with functionalized MWCNT. It was found to be toxic and induced massive loss of cell viability through DNA damage and programmed cell death of all doses compared to control. Our results demonstrate that carbon nanotubes indeed can be very toxic at sufficiently high concentrations from environmental and occupational exposure and that careful monitoring of toxicity studies is essential for risk assessment. PMID- 20016929 TI - Selenium restores defective beta-adrenergic receptor response of thoracic aorta in diabetic rats. AB - Increased oxidative stress is one of the basic contributors to the development of the cardiovascular complications in diabetes. Both endothelial and vascular smooth muscle cell dysfunctions are the main sign involved in the pathogenesis of diabetic cardiovascular dysfunction. Matrix metalloproteinases (MMPs) are expressed in the vasculature, and participate in tissue remodeling under pathological conditions such as increased oxidative stress, whereas little is known about effect of hyperglycemia on regulation of MMPs in vascular system. Therefore, we aimed to evaluate the effect of an antioxidant, sodium selenate treatment (0.3 mg/kg for 4 weeks) on function of streptozotocin-diabetic rat aorta. Sodium selenate treatment improved significantly impaired isoproterenol induced relaxation responses and contraction responses of the aortic strips, and exhibited marked protection against diabetes-induced degenerative changes in the smooth muscle cell morphology. Biochemical data showed that sodium selenate treatment induced a significant regulation of MMP-2 activity and protein loss as well as normalization of increased levels of tissue nitrite and protein thiol oxidation. In addition, this treatment restored diabetes-induced increased levels of endothelin-1, PKC, and cAMP production in the aortic tissue. Taken together, our data demonstrate that these beneficial effects of sodium selenate treatment in diabetics are related to be not only inhibition of increased oxidative stress but also prevention of both receptor- and smooth muscle-mediated dysfunction of vasculature, in part, via regulation of MMP-2. Such an observation provides evidence for potential therapeutic usage of selenium compounds for the amelioration of vascular disorders in diabetes. PMID- 20016930 TI - Elemol and amyris oil repel the ticks Ixodes scapularis and Amblyomma americanum (Acari: Ixodidae) in laboratory bioassays. AB - The essential oil from Amyris balsamifera (Rutaceae) and elemol, a principal constituent of the essential oil of Osage orange, Maclura pomifera (Moraceae) were evaluated in in vitro and in vivo laboratory bioassays for repellent activity against host-seeking nymphs of the blacklegged tick, Ixodes scapularis, and the lone star tick, Amblyomma americanum. Both bioassays took advantage of the tendency of these host-seeking ticks to climb slender vertical surfaces. In one bioassay, the central portion of a vertical strip of filter paper was treated with test solution and ticks placed or allowed to crawl onto the untreated lower portion. In the other bioassay, a strip of organdy cloth treated with test solution was doubly wrapped (treatment on outer layer) around the middle phalanx of a forefinger and ticks released on the fingertip. Both amyris oil and elemol were repellent to both species of ticks. Elemol did not differ significantly in effectiveness against A. americanum from the widely used repellent deet. At 2 and 4 h after application to filter paper, 827 microg amyris oil/cm(2) paper repelled 80 and 55%, respectively, of A. americanum nymphs. Ixodes scapularis was repelled by lower concentrations of amyris oil and elemol than A. americanum. PMID- 20016931 TI - A novel fluorescent silver ion biosensor based on nucleic acid molecular "light switch". AB - As one of nucleic acid molecular "light switch", Ru(bipy)(2)(dppx)(2+) is a good probe for the determination of double-helical DNA, which displays intense fluorescence when double-helical DNA is present. However, the fluorescence of Ru(bipy)(2)(dppx)(2+) is quenched when Ag(+) is added to the Ru(bipy)(2)(dppx)(2+)-DNA system. Based on the quenching of the fluorescence of Ru(bipy)(2)(dppx)(2+)-DNA system by Ag(+), a simple, rapid and specific method for Ag(+) determination was proposed. In the optimum conditions, Ag(+) concentration versus Ru(bipy)(2)(dppx)(2+) fluorescence intensity gave a linear response in the range from 0.2 to 6.0 microM with a detection limit (3sigma) of 3.2 x 10(-8) M. The proposed method has been applied to determine the Ag(+) in water samples and sulfadiazine silver cream successfully. Because of the intense fluorescence of Ru(bipy)(2)dppx(2+) when DNA is present, the interaction between Ag(+) and DNA was confirmed by fluorescence microscopy and the phenomenon of the fluorescence images agreed well with the results. The possible mechanism of the reaction was also discussed by circular dichroism spectra and isothermal titration calorimetry. PMID- 20016932 TI - A long wavelength excitable fluorophore; chloro phenyl imino propenyl aniline (CPIPA) for selective sensing of Hg (II). AB - In this study, a very sensitive and highly selective irreversible optical chemical sensor (optode) for mercury ions was described. The sensing scheme was based on the interaction of Hg (II) with a newly synthesized fluoroionophore; chloro phenyl imino propenyl aniline (CPIPA) in plasticized PVC membrane. The sensor membranes were tested for the determination of mercury ion in aqueous solutions by batch and flow-through methods. The optodes allow determination of Hg (II) in the working range of 1.0 x 10(-9)-1.0 x 10(-5) M with a detection limit of 4.3 ppb. The sensor exhibited excellent selectivity for Hg (II) with respect to several common alkali, alkaline earth and transition metal ions. The association constant of the 1:1 complex formation for Hg (II) was found to be K(a) = 1.86 x 10(5) M(-1). The CPIPA exhibited high fluorescence quantum yield, long excitation and emission wavelength and high Stokes' shift values in the solid matrix which makes it compatible with solid state optics. PMID- 20016933 TI - Inhibition of N-acetylgalactosamine 4-sulfate 6-O-sulfotransferase by beta-D-4-O sulfo-N-acetylgalactosaminides bearing various hydrophobic aglycons. AB - N-acetylgalactosamine 4-sulfate 6-O-sulfotransferase (GalNAc4S-6ST) transfers sulfate to position 6 of GalNAc(4SO4) residues of chondroitin sulfate to yield chondroitin sulfate E (CS-E). We have previously demonstrated that phenyl-beta-D GalNAc(4SO4) could serve as an acceptor for GalNAc4S-6ST, thereby inhibiting GalNAc4S-6ST competitively. In this paper we compared the inhibitory effects of various glycosides in which various hydrophobic aglycons were attached to D GalNAc(4SO4) via ss anomeric configuration. p-Nitrophenyl-beta-D-GalNAc(4SO4) and p-chlorophenyl-beta-D-GalNAc(4SO4) were stronger inhibitors than phenyl-beta-D GalNAc(4SO4). Among inhibitors examined here, 3-estradiol-beta-D-GalNAc(4SO4) was the strongest inhibitor; the Ki of 3-estradiol-beta-D-GalNAc(4SO4) for the competitive inhibition was 0.008 mM, which was much lower than the Ki of phenyl beta-D-GalNAc(4SO4), 0.98 mM. In contrast, 7-estradiol-beta-D-GalNAc(4SO4) showed only weak inhibition to GalNAc4S-6ST. 3-Estradiol-beta-D-GalNAc(4SO4) did not inhibit chondroitin 6-sulfotransferase and chondroitin 4-sulfotransferase under the concentration where GalNAc4S-6ST was inhibited by 90%. When 3-estradiol-beta D-GalNAc(4SO4) was added to the culture medium of chondrosarcoma cells expressing human GalNAc4S-6ST, a significant, albeit small, reduction in the cellular synthesis of CS-E was observed. These results suggest that estradiol group of 3 estradiol-beta-D-GalNAc(4SO4) may enhance the inhibitory activity of the glycoside through increasing the affinity to the enzyme and may allow the glycosides to diffuse at a low efficiency into the cells to inhibit cellular synthesis of CS-E. PMID- 20016934 TI - Flexibility in community pharmacy: a qualitative study of business models and cognitive services. AB - OBJECTIVE: To identify the capacity of current pharmacy business models, and the dimensions of organisational flexibility within them, to integrate products and services as well as the perceptions of viability of these models. METHODS: Fifty seven semi-structured interviews were conducted with community pharmacy owners or managers and support staff in 30 pharmacies across Australia. A framework of organisational flexibility was used to analyse their capacity to integrate services and perceptions of viability. Data were analysed using the method of constant comparison by two independent researchers. RESULTS: The study found that Australian community pharmacies have used the four types of flexibility to build capacity in distinct ways and react to changes in the local environment. This capacity building was manifested in four emerging business models which integrate services to varying degrees: classic community pharmacy, retail destination pharmacy, health care solution pharmacy and networked pharmacy. The perception of viability is less focused on dispensing medications and more focused on differentiating pharmacies through either a retail or services focus. Strategic flexibility appeared to offer pharmacies the ability to integrate and sustainably deliver services more successfully than other types, as exhibited by health care solution and networked pharmacies. CONCLUSION: Active support and encouragement to transition from being dependent on dispensing to implementing services is needed. The study showed that pharmacies where services were implemented and showed success are those strategically differentiating their businesses to become focused health care providers. This holistic approach should inevitably influence the sustainability of services. PMID- 20016935 TI - Ways of asking, ways of telling: a methodological comparison of ethnographic and research diagnostic interviews. AB - The interpretive understanding that can be derived from interviews is highly influenced by methods of data collection, be they structured or semistructured, ethnographic, clinical, life-history or survey interviews. This article responds to calls for research into the interview process by analyzing data produced by two distinctly different types of interview, a semistructured ethnographic interview and the Structured Clinical Interview for DSM, conducted with participants in the Navajo Healing Project. We examine how the two interview genres shape the context of researcher-respondent interaction and, in turn, influence how patients articulate their lives and their experience in terms of illness, causality, social environment, temporality and self/identity. We discuss the manner in which the two interviews impose narrative constraints on interviewers and respondents, with significant implications for understanding the jointly constructed nature of the interview process. The argument demonstrates both divergence and complementarity in the construction of knowledge by means of these interviewing methods. PMID- 20016936 TI - Utility of the twelve-item World Health Organization Disability Assessment Schedule II (WHO-DAS II) for discriminating depression "caseness" and severity in Spanish primary care patients. AB - PURPOSE: The 12-item WHO-DAS II was developed to assess the activity limitations and participation restrictions experienced by individuals irrespective of medical diagnosis. In this paper we examine the known-groups' validity of the instrument by evaluating its ability to discriminate between patients with/without major depression, patients with depression with/without medical comorbidity, and patients with depression with different depression severity. METHOD: The participants were 3,615 PC patients from 17 regions of Spain, with a first-time diagnosis of major depressive episode according to the general practitioner. The 12-item WHO-DAS II, the PHQ-9, and a chronic medical conditions checklist were administered during the consultation. RESULTS: The statistical analyses indicated that the 12-item WHO-DAS II was able to discriminate between patients with/without depression and between those with different depression severity. The ROC analysis revealed that with a cutoff score >or=50, the instrument correctly classified 70.4% of the sample (area under the ROC curve = .76; sensitivity = 71.4%; specificity = 67.6%). CONCLUSIONS: Overall, our results support the discriminant validity of the 12-item WHO-DAS II for major depression, being quite recommendable its use in epidemiological research. PMID- 20016937 TI - Non-specific phytohormonal induction of AtMYB44 and suppression of jasmonate responsive gene activation in Arabidopsis thaliana. AB - The Arabidopsis thaliana transcription factor gene AtMYB44 was induced within 10 min by treatment with methyl jasmonate (MeJA). Wound-induced expression of the gene was observed in local leaves, but not in distal leaves, illustrating jasmonate-independent induction at wound sites. AtMYB44 expression was not abolished in Arabidopsis mutants insensitive to jasmonate (coi1), ethylene (etr1), or abscisic acid (abi3-1) when treated with the corresponding hormones. Moreover, various growth hormones and sugars also induced rapid AtMYB44 transcript accumulation. Thus, AtMYB44 gene activation appears to not be induced by any specific hormone. MeJA-induced activation of jasmonate-responsive genes such as JR2, VSP, LOXII, and AOS was attenuated in transgenic Arabidopsis plants overexpressing the gene (35S:AtMYB44), but significantly enhanced in atmyb44 knockout mutants. The 35S:MYB44 and atmyb44 plants did not show defectiveness in MeJA-induced primary root growth inhibition, indicating that the differences in jasmonate-responsive gene expression observed was not due to alterations in the jasmonate signaling pathway. 35S:AtMYB44 seedlings exhibited slightly elevated chlorophyll levels and less jasmonate- induced anthocyanin accumulation, demonstrating suppression of jasmonate-mediated responses and enhancement of ABA mediated responses. These observations support the hypothesis of mutual antagonistic actions between jasmonate- and abscisic acid-mediated signaling pathways. PMID- 20016938 TI - Mining of serum glycoproteins by an indirect approach using cell line secretome. AB - Glycosylation is the most important and abundant post-translational modification in serum proteome. Several specific types of glycan epitopes have been shown to be associated with various types of disease. Direct analysis of serum glycoproteins is challenging due to its wide dynamic range. Alternatively, glycoproteins can be discovered in the secretome of model cell lines and then confirmed in blood. However, there has been little experimental evidence showing cell line secretome as a tractable target for the study of serum glycoproteins. We used a hydrazine-based glycocapture method to selectively enrich glycoproteins from the secretome of the breast cancer cell line Hs578T. A total of 132 glycoproteins were identified by nanoLC-MS/MS analysis. Among the identified proteins, we selected 13 proteins that had one or more N-glycosylation motifs in the matched peptides, which were included in the Secreted Protein Database but not yet in the Plasma Proteome Database (PPD), and whose antibodies were commercially available. Nine out of the 13 selected proteins were detected from human blood plasma by western analysis. Furthermore, eight proteins were also detected from the plasma by targeted LC-MS/MS, which had never been previously identified by data-dependent LC-MS/MS. Our results provide novel proteins that should be enrolled in PPD and suggest that analysis of cell line secretome with subfractionation is an efficient strategy for discovering disease-relevant serum proteins. PMID- 20016939 TI - Hoxc8 represses BMP-induced expression of Smad6. AB - Proper regulation of bone morphogenetic protein (BMP) signaling is critical for correct patterning and morphogenesis of various tissues and organs. A well known feedback mechanism is a BMP-mediated induction of Smad6, an inhibitor of BMP signaling. Hoxc8, one of the Hox family transcription factors, has also been shown to negatively regulate BMP-mediated gene expression. Here we add another level of Hoxc8 regulation on BMP signaling. Our results show that Hoxc8, when over-expressed in C3H10T1/2 or C2C12 cells, suppressed basal Smad6 promoter activity and its mRNA expression. Activation of Smad6 transcription either by BMP2 treatment or Smad1 over-expression was also abolished by Hoxc8. When chromatin was precipitated from mouse embryos with anti-Smad1 or anti-Hoxc8 antibody, Smad6 promoter sequence was enriched, suggesting that Hoxc8 proteins make complexes with Smad1 in the Smad6 promoter region. Yet, a lack of Hox binding motifs in the Smad6 promoter sequence suggests that instead of directly binding to the DNA, Hoxc8 may regulate Smad6 expression via an indirect mechanism. Our results suggest that the Smad6-mediated negative feedback mechanism on BMP signaling may be balanced by the repression of Smad6 expression by Hoxc8. PMID- 20016940 TI - Overexpression of BrMORN, a novel 'membrane occupation and recognition nexus' motif protein gene from Chinese cabbage, promotes vegetative growth and seed production in Arabidopsis. AB - Proteins that contain membrane occupation and recognition nexus (MORN) motifs regulate various aspects of cellular metabolism by localizing proteins in different cellular organelles. The full-length Brassica rapa MORN motif protein (BrMORN) cDNA consists of 1,510 bp encoding 502 deduced amino acids with a predicted molecular mass of 55.8 kDa and an isoelectric point of 9.72. BrMORN is a novel protein composed of two N-terminal transmembrane helices and seven C terminal MORN motifs and it appears to be localized on the plastid envelope. BrMORN expression was relatively high in actively-growing tissues, but low in mature tissues and under some abiotic stresses. Arabidopsis thaliana plants overexpressing BrMORN showed an enhanced rate of growth, hypocotyl elongation, and increases in the size of vegetative organs and seed productivity under normal growth conditions. In addition, cell size in Arabidopsis plants overexpressing BrMORN was 24% larger than that of wild-type plants, implying that the increase in the size of vegetative organs is due to cell enlargement. The increased size of the vegetative organs also led to increased seed production. Our data suggest that the MORN motif of BrMORN may act at the plastid envelope and facilitate plant growth via cell enlargement. PMID- 20016941 TI - Identification of an Arabidopsis Nodulin-related protein in heat stress. AB - We identified a Nodulin-related protein 1 (NRP1) encoded by At2g03440, which was previously reported to be RPS2 interacting protein in yeast-two-hybrid assay. Northern blotting showed that AtNRP1 expression was suppressed by heat stress (42 degrees C) and induced by low temperature (4 degrees C) treatment. Strong GUS staining was observed in the sites of meristematic tissues of pAtNRP1:: GUS transgenic plants, such as shoot apex and root tips, young leaf veins, stamens and stigmas of flowers, and abscission layers of young siliques. To study AtNRP1 biological functions, we have characterized both loss-of-function T-DNA insertion and transgenic overexpression plants for AtNRP1 in Arabidopsis. The T-DNA insertion mutants displayed no obvious difference as compared to wild-type Arabidopsis under heat stress, but the significant enhanced susceptibility to heat stress was revealed in two independent AtNRP1-overexpressing transgenic lines. Further study found that the decreased thermotolerance in AtNRP1 overexpressing lines accompanied significantly decreased accumulation of ABA after heat treatment, which was probably due to AtNRP1 playing a role in negative feedback regulation of the ABA synthesis pathway. These results support the viewpoint that the application of ABA inhibits nodulation and nodulin-related gene expression and threaten adverse ambient temperature can impact the nodulin related gene expression. PMID- 20016942 TI - IFN-gamma down-regulates TGF-beta1-induced IgA expression through Stat1 and p300 signaling. AB - IFN-gamma has been shown to either up- or down-regulate the expression of specific TGF-beta1-induced target genes. We investigated the effect of IFN-gamma on TGF-beta1-induced IgA isotype expression. We found that IFN-gamma inhibited not only TGF-beta1-induced germ-line (GL) alpha transcription, but also IgA secretion by TGF-beta1-stimulated murine B cells. Overexpression of Stat1 diminished TGF-beta1-induced, Smad3/4-and Runx3-mediated GL alpha promoter activity. Overexpression of p300 also increased the promoter activity, while its effect was abrogated by co-transfected Stat1. Stat1 interfered with the Smad3:p300 interaction, likely due to a stronger Stat1:p300 binding affinity. These results indicate that Stat1 can inhibit GL alpha transcription through binding to p300. Further, overexpression of SOCS1, a JAK inhibitor, diminished the antagonistic effect of IFN-gamma on TGF-beta1-induced GL alpha transcription and IgA secretion. These results indicate that JAK/Stat1-mediated IFN-gamma signaling antagonizes TGF-beta1-induced GL alpha transcription, mainly through deprivation of p300 from Smad3, resulting in decreased IgA synthesis. PMID- 20016943 TI - Mycobacterium tuberculosis-induced expression of Leukotactin-1 is mediated by the PI3-K/PDK1/Akt signaling pathway. AB - Chemokines function in the migration of circulating leukocytes to regions of inflammation, and have been implicated in chronic inflammatory conditions including mycobacterial infection. We investigated whether Leukotactin-1 (Lkn-1), a novel member of the CC-chemokines, is involved in the immune response of macrophages against Mycobacterium tuberculosis (MTB). In PMA-differentiated THP-1 cells, MTB infection increased mRNA expression of Lkn-1 in a dose-dependent manner. Lkn-1 induction peaked 12 h after infection, then declined gradually and returned to its basal level at 72 h. Secretion of Lkn-1 was elevated by MTB infection. The increase in expression and secretion of Lkn-1 caused by MTB was reduced in cells treated with inhibitors of phosphatidylinositol 3-kinase (PI3 K), 3-phosphoinositide-dependent kinase 1 (PDK1) and Akt. MTB-induced Akt phosphorylation was blocked by treatment with a PI3-K inhibitor or a PDK1 inhibitor, implying that PI3-K, PDK1, and Akt are associated with the signaling pathway that up-regulates Lkn-1 in response to MTB. These results suggest that Lkn-1 is novel member of the group of chemokines that is released by macrophages infected with MTB. PMID- 20016944 TI - Identification of single nucleotide polymorphisms in the TNFRSF17 gene and their association with gastrointestinal disorders. AB - TNFRSF17 is preferentially expressed in mature B lymphocytes, and may be important for the development of B cells. TNFRSF17 is selected as a candidate susceptibility gene to IBD pathogenesis by our cDNA microarray analysis, and we showed the specific expression of TNFRSF17 in resting and activated CD19(+) cells obtained from human blood. We identified four SNPs (g-1729G>A, g.2295T>C, g.2445G>A and g.2493G>A) and one variation site (g.894delT) in the TNFRSF17 gene using direct sequencing analysis. In addition, the association of the genotype and allelic frequencies of these SNPs was studied in healthy controls and in patients with ulcerative colitis (UC) or irritable bowel syndrome (IBS). Although, the genotype and allelic frequencies of these SNPs, in the UC and IBS patients, were not significantly different from those in the healthy controls, the distribution of the AAG, GGA, AGG and AAA haplotypes, of the SNPs (g. 1729G>A, g.2445G> A and g.2493G>A) associated with the TNFRSF17 gene, in the UC patients, were notably different from those of the healthy controls (P = 0.002, 0.002, 4.7E-4 and 3.3E-6, respectively). Moreover, the frequencies of the AAG, AGG, GAG and GAA haplotypes were significantly different in the IBS patients compared to the healthy controls (P = 4.2E-5, 4.4E-17, 1.8E-22 and 1.6E-10, respectively). These results suggest that the haplotypes of the TNFRSF17 polymorphisms might be associated with UC and IBS susceptibility. PMID- 20016945 TI - Silencing of NbCEP1 encoding a chloroplast envelope protein containing 15 leucine rich-repeats disrupts chloroplast biogenesis in Nicotiana benthamiana. AB - We characterized the physiological functions of Nicotiana benthamiana Chloroplast Envelope Protein 1 (NbCEP1) in Nicotiana benthamiana. NbCEP1 contains a chloroplast transit peptide and a single transmembrane domain at the N terminus, and most of its protein coding region is comprised of 15 leucine-rich-repeats (LRRs). The NbCEP1 gene is expressed in both aerial and underground plant tissues, and is induced by light. A GFP fusion protein of full length NbCEP1 was targeted to the chloroplast envelope and co-localized with OEP7:RFP, a marker protein for the chloroplast envelope. A fusion protein consisting of GFP and the NbCEP1 transit peptide mainly localized in the chloroplast stroma. Reduction of NbCEP1 expression by virus-induced gene silencing resulted in a leaf yellowing phenotype without much affecting overall plant growth. At the cellular level, depletion of NbCEP1 severely influenced chloroplast development, reducing both the number and size of the chloroplasts. Interestingly, mitochondrial development was also impaired, possibly an indirect effect of chloroplast ablation. A deficiency in NbCEP1 activity decreased the chlorophyll and carotenoid levels. Our results suggest that NbCEP1 plays a critical function, possibly through protein-protein interactions mediated by its LRRs, in chloroplast development in N. benthamiana. PMID- 20016946 TI - Fine mapping and candidate gene analysis of the floury endosperm gene, FLO(a), in rice. AB - In addition to its role as an energy source for plants, animals and humans, starch is also an environmentally friendly alternative to fossil fuels. In rice, the eating and cooking quality of the grain is determined by its starch properties. The floury endosperm of rice has been explored as an agronomical trait in breeding and genetics studies. In the present study, we characterized a floury endosperm mutant, flo(a), derived from treatment of Oryza sativa ssp. japonica cultivar Hwacheong with MNU. The innermost endosperm of the flo(a) mutant exhibited floury characteristics while the outer layer of the endosperm appeared normal. Starch granules in the flo(a) mutant formed a loosely-packed crystalline structure and X-ray diffraction revealed that the overall crystallinity of the starch was decreased compared to wild-type. The FLO(a) gene was isolated via a map-based cloning approach and predicted to encode the tetratricopeptide repeat domain-containing protein, OsTPR. Three mutant alleles contain a nucleotide substitution that generated one stop codon or one splice site, respectively, which presumably disrupts the interaction of the functionally conserved TPR motifs. Taken together, our map-based cloning approach pinpointed an OsTPR as a strong candidate of FLO(a), and the proteins that contain TPR motifs might play a significant role in rice starch biosynthetic pathways. PMID- 20016947 TI - Mesenchymal stem cells modified with stromal cell-derived factor 1 alpha improve cardiac remodeling via paracrine activation of hepatocyte growth factor in a rat model of myocardial infarction. AB - Mesenchymal stem cells (MSCs) are a promising source for cell-based treatment of myocardial infarction (MI), but existing strategies are restricted by low cell survival and engraftment. We examined whether SDF-1 transfection improve MSC viability and paracrine action in infarcted hearts. We found SDF-1-modified MSCs effectively expressed SDF-1 for at least 21 days after exposure to hypoxia. The apoptosis of Ad-SDF-1-MSCs was 42% of that seen in Ad-EGFP-MSCs and 53% of untreated MSCs. In the infarcted hearts, the number of DAPI-labeling cells in the Ad-SDF-1-MSC group was 5-fold that in the Ad-EGFP-MSC group. Importantly, expression of antifibrotic factor, HGF, was detected in cultured MSCs, and HGF expression levels were higher in Ad-SDF-MSC-treated hearts, compared with Ad-EGFP MSC or control hearts. Compared with the control group, Ad-SDF-MSC transplantation significantly decreased the expression of collagens I and III and matrix metalloproteinase 2 and 9, but heart function was improved in d-SDF-MSC treated animals. In conclusion, SDF-1-modified MSCs enhanced the tolerance of engrafted MSCs to hypoxic injury in vitro and improved their viability in infarcted hearts, thus helping preserve the contractile function and attenuate left ventricle (LV) remodeling, and this may be at least partly mediated by enhanced paracrine signaling from MSCs via antifibrotic factors such as HGF. PMID- 20016948 TI - Capacitive facial movement detection for human-computer interaction to click by frowning and lifting eyebrows: assistive technology. AB - A capacitive facial movement detection method designed for human-computer interaction is presented. Some point-and-click interfaces use facial electromyography for clicking. The presented method provides a contactless alternative. Electrodes with no galvanic coupling to the face are used to form electric fields. Changes in the electric fields due to facial movements are detected by measuring capacitances between the electrodes. A prototype device for measuring a capacitance signal affected by frowning and lifting eyebrows was constructed. A commercial integrated circuit for capacitive touch sensors is used in the measurement. The applied movement detection algorithm uses an adaptive approach to provide operation capability in noisy and dynamic environments. Experimentation with 10 test subjects proved that, under controlled circumstances, the movements are detected with good efficiency, but characterizing the movements into frowns and eyebrow lifts is more problematic. Integration with a two-dimensional (2D) pointing solution and further experiments are still required. PMID- 20016949 TI - Ambulatory human motion tracking by fusion of inertial and magnetic sensing with adaptive actuation. AB - Over the last years, inertial sensing has proven to be a suitable ambulatory alternative to traditional human motion tracking based on optical position measurement systems, which are generally restricted to a laboratory environment. Besides many advantages, a major drawback is the inherent drift caused by integration of acceleration and angular velocity to obtain position and orientation. In addition, inertial sensing cannot be used to estimate relative positions and orientations of sensors with respect to each other. In order to overcome these drawbacks, this study presents an Extended Kalman Filter for fusion of inertial and magnetic sensing that is used to estimate relative positions and orientations. In between magnetic updates, change of position and orientation are estimated using inertial sensors. The system decides to perform a magnetic update only if the estimated uncertainty associated with the relative position and orientation exceeds a predefined threshold. The filter is able to provide a stable and accurate estimation of relative position and orientation for several types of movements, as indicated by the average rms error being 0.033 m for the position and 3.6 degrees for the orientation. PMID- 20016950 TI - Reexamination of risk criteria in dengue patients using the self-organizing map. AB - Even though the World Health Organization criteria's for classifying the dengue infection have been used for long time, recent studies declare that several difficulties have been faced by the clinicians to apply these criteria. Accordingly, many studies have proposed modified criteria to identify the risk in dengue patients based on statistical analysis techniques. None of these studies utilized the powerfulness of the self-organized map (SOM) in visualizing, understanding, and exploring the complexity in multivariable data. Therefore, this study utilized the clustering of the SOM technique to identify the risk criteria in 195 dengue patients. The new risk criteria were defined as: platelet count less than or equal 40,000 cells per mm(3), hematocrit concentration great than or equal 25% and aspartate aminotransferase (AST) rose by fivefold the normal upper limit for AST/alanine aminotransfansferase (ALT) rose by fivefold the normal upper limit for ALT. The clusters analysis indicated that any dengue patient fulfills any two of the risk criteria is consider as high risk dengue patient. PMID- 20016951 TI - From the editors' desk. PMID- 20016952 TI - Assessment of the contributions of clinician educators. PMID- 20016953 TI - Introducing exercises in clinical reasoning. PMID- 20016954 TI - Medicare Part D--lessons learned and guidance for health-care reform. PMID- 20016955 TI - Caring for Ivan Ilyich. AB - For over a century, Leo Tolstoy's The Death of Ivan Ilych has been one of the most influential examinations of how we come to terms with our own mortality. Of the many who care for Ivan Ilych, only the uneducated peasant, Gerasim, is able to help him find meaning and resolution before death. An excerpt that describes Gerasim's key interaction with Ivan Ilych is provided. Analysis of the text reveals how cultural values may hinder a patient's ability to confront mortality and how unique social barriers inhibit different caretakers' ability to care for a dying patient. PMID- 20016956 TI - Juvenile osteochondritis of femoral condyles: treatment with transchondral drilling. Analysis of 40 cases. AB - INTRODUCTION: Osteochondritis dissecans (OCD) of the femoral condyle is a rare lesion. MATERIALS AND METHODS: A retrospective study (level IV evidence) analyzing a series of 40 pediatric cases with juvenile femoral condyles osteochondritis treated by arthroscopic multiple transchondral drilling between February 1999 and June 2008 was undertaken. This lesion affected the medial condyle in 87.5% of cases. The average age at treatment was 13.4 years. Our study took into account the location of the lesion and its radiological evolutionary stage. The average follow up was 14.8 months. The postoperative evaluation was based on the clinical and radiological scores of Hughston. RESULTS: Good clinical and radiological results in 97.5 and 95% of cases, respectively were obtained, with a significant correlation (P < 0.001) between clinical scores and radiological Hughston scores. The closed nature of the growth plate during surgery has a significant deleterious effect (P < 0.001) on the clinical and radiological score of Hughston. CONCLUSION: All patients presenting juvenile condylar osteochondritis with open growth plate during treatment had good clinical and radiological results, confirming the validity and effectiveness of multiple transchondral drilling in this type of lesion. PMID- 20016958 TI - Capsaicin receptors are colocalized with sweet/bitter receptors in the taste sensing cells of circumvallate papillae. AB - We examined co-localization of vanilloid receptor (VR1) with sweet receptors T1R2, T1R3, or bitter receptor T2R6 in taste receptor cells of rat circumvallate papillae. Tissue sections of rat circumvallate papillae were doubly reacted with anti-VR1 antibodies and anti-T1R2, anti-T1R3 or anti-T2R6 antibodies, using double-immunofluorescence histochemistry technique. Localizations of VR1, T1Rs and T2R6 in the vallate taste cells containing alpha-gustducin were also examined. VR1 immunoreactivities (-ir) were observed in subsets of taste cells in the circumvallate papillae, and 96-99% of the vallate taste cells exhibiting T1R2 , T1R3- or T2R6-ir co-exhibited VR1-ir. Approximately half of T2R6-ir cells (~49%), and 50-58% of T1Rs-ir cells, co-exhibited alpha-gustducin-ir in the vallate taste buds. About 58% of VR1-ir cells in the vallate exhibited alpha gustducin-ir as well. Results support the idea that capsaicin may interact with the transduction pathways of sweet and bitter taste stimuli, possibly in mediation of its receptor VR1 localized in taste receptor cells. Additionally, the partial co-localization of alpha-gustducin with VR1 suggests that a tentative modulatory function of capsaicin in sweet and bitter transductions in the rat circumvallate comprises of both alpha-gustducin-mediated and non-mediated transduction pathways. PMID- 20016957 TI - Consensus guidance for banking and supply of human embryonic stem cell lines for research purposes. PMID- 20016959 TI - History of blood transfusion before 1990 is associated with increased risk for cancer mortality independently of liver disease: a prospective long-term follow up study. AB - OBJECTIVES: The aim of this work is to investigate the association between transfusion history and cancer mortality in a prospective follow-up study. METHODS: We conducted a prospective cohort study in four areas of Akita Prefecture, Japan, in 10,451 individuals (4,401 men and 6,050 women, aged 40-79 years) without history of cancer. The subjects were followed until 31 December 2003 and the number of deaths from cancer was recorded. RESULTS: After mean follow-up of 12.76 years (140,259 person-years), 520 individuals (333 men and 187 women) died of cancer. History of blood transfusion before 1990 was mildly but significantly associated with overall cancer mortality (hazard ratio = 1.75, 95% confidence interval: 1.32-2.18) and nonliver cancer mortality (HR = 1.68, 95% CI: 1.25-2.26). This significant association remained unchanged after excluding deaths that occurred within 5 years of baseline for overall cancer mortality (HR = 1.47, 95% CI: 1.04-2.09) and for nonliver cancer mortality (HR = 1.43, 95% CI: 1.00-2.04). The significant association for nonliver cancer mortality was confirmed in subjects with no smoking history and/or alcohol consumption (HR = 2.01, 95% CI: 1.35-3.00). Site-specific analysis showed a possible association between transfusion history and death from pancreatic cancer. CONCLUSIONS: History of blood transfusion before 1990 was found to be associated with increased risk for cancer mortality and was independent of liver diseases. The mechanism of the association between blood transfusion and cancer mortality warrants further research. PMID- 20016960 TI - Pheno- and genotypic features of Epstein-Barr virus associated B-cell lymphoproliferations in peripheral T-cell lymphomas. AB - Among the 300 peripheral T-cell lymphomas (PTCL) searched for EBV positive non resting B-cells by EBER in situ hybridization 12 have been identified with various forms of EBV-driven B-cell proliferation. This could be categorized into three major forms. i. In the first form scattered immature, mononuclear B-cells of immuno-, centroblastic type with CD20+. CD30+ CD45+, LMP1+ phenotype, reactive appearance and polyclonal immunoglobulin heavy chains gene rearrangement (IgH-R) were admixed to the PTCL cells. ii. The second form mimicked diffuse large B-cell lymphoma as homogenous sheets, largely demarcated from the PTCL, of mononuclear, immature B-cell of CD20+, CD30+, CD45+, LMP1+, EBNA-2+ phenotype but with lack of monoclonal IgH-R were present. iii. In the third form scattered Hodgkin-Reed Sternberg (HRS) type of cells were noticed which exhibited the CD15+/-, CD20-/+, CD30+, CD45-, LMP1+, EBNA-2- phenotype and in 50% showed clonal IgH gene rearrangement in whole tissue DNA extract. The IgH associated transcription factors' (OCT2, BOB.1/OBF.1, PU.1) expression patterns in these cells corresponded to those of HRS cells in cHL. Based on analysis of 65 PTCLs, we have identified in the positive cases a highly significant increase of EBV+ small, reactive, resting B-cell compartment (75.9 / 100 HPF in PTCL vs. 1.5 / 100 HPF in control lymph nodes) likely to be due to the decreased immune surveillance. This progressive accumulation of EBV+ by-stander B-cell population in PTCLs might be the source of various B-cell proliferations, which in any form represent major diagnostic pitfalls and require a careful differential diagnostic procedure. PMID- 20016961 TI - Vasculogenic mimicry: a new prognostic sign of gastric adenocarcinoma. AB - Vasculogenic mimicry (VM) has been generally recognized as a new pattern of tumor neovascularization. It presents in many human malignancies. Till now, there is no report about VM in gastric adenocarcinoma (GAC). In this study, we collected 173 paraffin-embedded human GAC samples, with detailed follow-up and clinicopathologic data. CD31/ periodic acid-Schiff (PAS) double staining, immunohistochemical staining of CK8 & 18 and laminin were performed to validate the existence of VM in GAC. Microvascular density (MVD) and vasulogenic mimicry density (VMD) were counted respectively. VM was observed in 40 of the 173 GAC patients, especially in poorly differentiated GAC (P = 0.014). Patients with VM were prone to hematogenous metastasis and distant recurrence compared with patients without VM (P = 0.020, 0.029). Higher VMD values was also associated with hematogenous metastasis (P = 0.003). Immunohistochemical staining index (SI) of hypoxia-inducible factor 1alpha (HIF-1alpha), vascular endothelial growth factor (VEGF), matrix metalloproteinase (MMP)-2, and MMP-9 were compared between the VM and non-VM group. The SI of four factors were all higher in the VM group than those of non-VM group (P = 0.000, 0.000, 0.004, 0.009, respectively). The Kaplan-Meier survival analysis showed that the VM group has shorter life span compared with non-VM group (P = 0.022). Cox proportional hazards model indicated that the presence of VM and TNM stage were independent predictors of poor prognosis (P = 0.039 and 0.004) for GAC. In conclusion, VM exists in GAC, especially in poorly differentiated GAC. Additionally, it is an unfavorable prognostic indictor for GAC. Hypoxia may play a role in VM formation in GAC. PMID- 20016962 TI - Proliferating activity in paget disease of the nipple. AB - Paget disease of the nipple is a rare disease characterized by the presence of malignant glandular cells within the squamous epithelium of the nipple. The most common hypothesis to explain the development of Paget disease is an intraepithelial epidermotropic migration of malignant epithelial cells originating from an underlying intraductal carcinoma. If the immunohistochemical properties of the Paget cells in the nipple have been extensively studied, their proliferating characteristics remain paradoxically poorly studied. In the present study we have investigated the proliferating activity of Paget cells in the nipple by using double stain immunohistochemistry with both Ki67 (a protein which is expressed in all active parts of the cell cycle) and cytokeratin 7 (a highly sensitive marker of Paget cells). Ten cases of Paget disease and in their associated intraductal carcinomas (n = 10) and/or invasive carcinomas (n = 4) were tested. The mean Ki67 index was in Paget disease (26% +/- 10), in intraductal carcinomas (23% +/- 8) and/or in invasive carcinomas (20% +/- 8) (p > 0,05). This is the first report to convincingly demonstrate by specific double stain immunohistochemistry that Paget disease and underlying intraductal carcinomas share a close proliferating activity. PMID- 20016963 TI - Structural changes associated with progression of motor deficits in spinocerebellar ataxia 17. AB - Spinocerebellar ataxia (SCA17) is a rare genetic disorder characterized by a variety of neuropsychiatric symptoms. Recently, using magnetic resonance imaging (MRI) voxel-based morphometry (VBM), several specific functional-structural correlations comprising differential degeneration related to motor and psychiatric symptoms were reported in patients with SCA17. To investigate gray matter volume (GMV) changes over time and its association to clinical neuropsychiatric symptomatology, nine SCA17 mutation carriers and nine matched healthy individuals underwent a detailed neuropsychiatric clinical examination and a high-resolution T1-weighted volume MRI scan, both at baseline and follow-up after 18 months. Follow-up images revealed a progressive GMV reduction in specific degeneration patterns. In contrast to healthy controls, SCA17 patients showed a greater atrophy not only in cerebellar regions but also in cortical structures such as the limbic system (parahippocampus, cingulate) and parietal precuneus. Clinically, progression of motor symptoms was more pronounced than that of psychiatric symptoms. Correlation with the clinical motor scores revealed a progressive reduction of GMV in cerebellar and cerebral motor networks, whereas correlation with psychiatric scores displayed a more widespread GMV impairment in frontal, limbic, parietal, and also cerebellar structures. Interestingly, changes in global functioning were correlated with bilateral atrophy within the para /hippocampus. While there was a good temporal association between worsening of motor symptoms and progression in cerebral and cortical neurodegeneration, the progression in psychiatric related neurodegeneration seemed to be more widespread and complex, showing progressive atrophy that preceded the further development of clinical psychiatric symptoms. PMID- 20016964 TI - Multiunit recording of the cerebellar cortex, inferior olive, and fastigial nucleus during copulation in naive and sexually experienced male rats. AB - The sexual behavior of male rats constitutes a natural model to study learning of motor skills at the level of the central nervous system. We previously showed that sexual behavior increases Fos expression in granule cells at lobules 6 to 9 of the vermis cerebellum. Herein, we obtained multiunit recordings of lobules 6a and 7 during the training period of naive subjects, and during consecutive ejaculations of expert males. Recordings from both lobules and the inferior olive showed that the maximum amplitude of mount, intromission, and ejaculation signals were similar, but sexual behavior during training tests produced a decrease in the amplitude for mount and intromission signals. The fastigial nucleus showed an inverse mirror-like response. Thus, the cerebellum is involved in the neural basis of sexual behavior and the learning of appropriate behavioral displays during copulation, with a wiring that involves the cerebellar cortex, inferior olive, and fastigial nucleus. PMID- 20016965 TI - Establishment and characterization of human osteosarcoma cell lines with different pulmonary metastatic potentials. AB - To develop an investigative tool for the study of human osteosarcoma (OSA), we established a human OSA cell line, namely, SOSP-9607, which exhibits a potential for spontaneous pulmonary metastasis. Subsequently, we screened two related sublines (F5M2 and F4) that have different pulmonary metastatic potentials. An in vivo orthotopic transplantation assay confirmed spontaneous pulmonary metastasis in all mice (100%) transplanted with the more aggressive OSA cells (F5M2) and a lesser degree of metastases with smaller nodules in 33.3% mice transplanted with the less aggressive OSA cell subline (F4). In mice transplanted with F5M2 cells, death from metastasis occurred at a median of 71 days; however, in mice transplanted with F4, no death occurred even after 120 days. Therefore, the F5M2 and F4 sublines, which originated from the same parent cell line, differed with respect to metastasis-related properties such as proliferating ability and invasiveness. Hence, these well-characterized human OSA sublines can be used as valuable models for comparative studies of genetic determinants of OSA in the future. PMID- 20016966 TI - Tissue and host tropism of influenza viruses: importance of quantitative analysis. AB - It is generally accepted that human influenza viruses preferentially bind to cell surface glycoproteins/glycolipids containing sialic acids in alpha2,6-linkage; while avian and equine influenza viruses preferentially bind to those containing sialic acids in alpha2,3-linkage. Even though this generalized view is accurate for H3 subtype isolates, it may not be accurate and absolute for all subtypes of influenza A viruses and, therefore, needs to be reevaluated carefully and realistically. Some of the studies published in major scientific journals on the subject of tissue tropism of influenza viruses are inconsistent and caused confusion in the scientific community. One of the reasons for the inconsistency is that most studies were quantitative descriptions of sialic acid receptor distributions based on lectin or influenza virus immunohistochemistry results with limited numbers of stained cells. In addition, recent studies indicate that alpha2,3- and alpha2,6-linked sialic acids are not the sole receptors determining tissue and host tropism of influenza viruses. In fact, determinants for tissue and host tropism of human, avian and animal influenza viruses are more complex than what has been generally accepted. Other factors, such as glycan topology, concentration of invading viruses, local density of receptors, lipid raft microdomains, coreceptors or sialic acid-independent receptors, may also be important. To more efficiently control the global spread of pandemic influenza such as the current circulating influenza A H1N1, it is crucial to clarify the determinants for tissue and host tropism of influenza viruses through quantitative analysis of experimental results. In this review, I will comment on some conflicting issues related to tissue and host tropism of influenza viruses, discuss the importance of quantitative analysis of lectin and influenza virus immunohistochemistry results and point out directions for future studies in this area, which should lead to a better understanding of tissue and host tropism of influenza viruses. PMID- 20016967 TI - Mechanisms of microRNA-mediated gene regulation. AB - microRNAs (miRNAs) are identified as a class of non-protein regulators and a new source for broad control of gene expression in eukaryotes. The past years have witnessed substantial progress in understanding miRNA functions and mechanisms, although a few controversies remain. Various hypotheses and models have been suggested for the mechanisms of miRNA repression, including translational inhibition at the level of initiation or elongation, rapid degradation of the nascent peptide, mRNA degradation, and mRNA sequestration into P bodies (processing bodies) and SGs (stress granules) for degradation or/and storage. Recently, some noncanonical miRNA regulation, such as miRNA activation and de repression of miRNA inhibition, have been uncovered. This review discusses some recent advances about how miRNAs regulate their targets and various modes of miRNA function. PMID- 20016968 TI - Circulating miRNA and cancer diagnosis. AB - miRNAs are a class of small RNA molecules with regulatory function, and play an important role in tumor development and progression. It has been demonstrated that tumor-derived miRNAs exist in the circulating nucleic acids of cancer patients. This phenomenon implies that detection of the circulating miRNA may be an effective method for non-invasive diagnosis of cancer. In this review, we summarize the applications of the circulating miRNA as biomarkers in cancer diagnosis, as well as the latest research progress in this area. PMID- 20016969 TI - Progress in miRNA target prediction and identification. AB - Recently, the identification of miRNA targets has received much attention. The strategies to determine miRNA targets include bioinformatic prediction and experimental assays. The bioinformatic prediction methods are mainly based on the confirmed rules of interaction between miRNAs and their targets, and are carried out by programs, such as miRanda, TargetScan, TargetScanS, RNAhybrid, DIANA microT, PicTar, RNA22 and FindTar, which follow well-known principles. The experimental assays to find miRNA targets employ immunoprecipitation of AGO proteins to identify interacting mRNAs, or the analysis of mRNA or protein levels to identify genes which can be regulated by miRNAs. The improvement of current bioinformatic and experimental assays and the development of novel assays will enable greater efficiency in the identification of miRNA targets and thus facilitate miRNA research. This paper describes progress in the prediction and identification of miRNA targets. PMID- 20016970 TI - Expression of hIGF-I in the silk glands of transgenic silkworms and in transformed silkworm cells. AB - To express human insulin-like growth factor-I (hIGF-I) in transformed Bombyx mori cultured cells and silk glands, the transgenic vector pigA3GFP-hIGF-ie-neo was constructed with a neomycin resistance gene driven by the baculovirus ie-1 promoter, and with the hIGF-I gene under the control of the silkworm sericin promoter Ser-1. The stably transformed BmN cells expressing hIGF-I were selected by using the antibiotic G418 at a final concentration of 700-800 microg/mL after the BmN cells were transfected with the piggyBac vector and the helper plasmid. The specific band of hIGF-I was detected in the transformed cells by Western blot. The expression level of hIGF-I, determined by ELISA, was about 7800 pg in 5x10(5) cells. Analysis of the chromosomal insertion sites by inverse PCR showed that exogenous DNA could be inserted into the cell genome randomly or at TTAA target sequence specifically for piggyBac element transposition. The transgenic vector pigA3GFP-hIGF-ie-neo was transferred into the eggs using sperm-mediated gene transfer. Finally, two transgenic silkworms were obtained after screening for the neo and gfp genes and verified by PCR and dot hybridization. The expression level of hIGF-I determined by ELISA was about 2440 pg/g of silk gland of the transgenic silkworms of the G1 generation. PMID- 20016971 TI - Temporal allocation of metabolic tolerance in the body of beet armyworm in response to three gossypol-cotton cultivars. AB - The nutrient composition and enzyme activities in larvae of the beet armyworm, Spodoptera exigua (Hubner), fed on high, medium or low gossypol cotton cultivars were examined at different time intervals. Significantly lower free fatty acid was observed in larvae fed for 6 h on high gossypol 'M9101' compared to larvae fed on the low (ZMS13) and intermediate (HZ401) gossypol cultivars. Significantly higher trypsin activity was observed in larvae fed on high gossypol 'M9101' for 24 h compared to those fed for 1, 4 and 6 h. Significantly higher catalase and total superoxide dismutase enzyme activities were observed in larvae of S. exigua fed on high gossypol 'M9101' compared with low gossypol cultivars 'ZMS13' and 'HZ401' for 1, 4, 6 and 24 h. However, significantly lower carboxylesterase and acetylcholinesterase enzyme activities were found in larvae fed on high gossypol 'M9101' compared with the other cultivars for 1, 4, 6 and 24 h. The interaction between cotton variety and beet armyworm infestation time significantly affected the carboxylesterase enzyme activity in S. exigua. The characterization of the effects of plant allelochemicals on herbivorous larvae is important for aiding understanding of plant-insect interaction as well as in devising solutions to pest problems by breeding plant resistance, identifying metabolic targets for insecticide development, etc. PMID- 20016972 TI - Characterization of interspecific hybrids and backcross progenies from a cross between Oryza minuta and Oryza sativa. AB - Oryza minuta, a tetraploid wild relative of cultivated rice, is an important source for the genetic improvement. Interspecific hybrids were obtained from the cross of O. sativa L. (IR24) and O. minuta (Acc. No. 101133) with 5.58% crossability, which ranged from 0.11% to 1.62% in the backcross generations. The chromosome numbers of the backcross progenies were 24 to 48. Seven yield-related traits of the parents, hybrid F(1), and backcross progenies were evaluated. Simple sequence repeat markers analysis showed that the polymorphism ratio of SSR bands between IR24 and Acc. No. 101133 was 93.2%. The average donor segment number, length, donor genome size, and percentage of donor genome of 92 BC(3)F(1) plants (2n=24) were 24.1, 17.8 cM, 438.4 cM and 26.2%, respectively. They were complex variation and uneven among the chromosomes. These introgression lines could be used to identify the favorable genes of O. minuta and provide a new platform for the genetic improvement of cultivated rice. PMID- 20016973 TI - Silence of Synaptotagmin VII inhibits release of dense core vesicles in PC12 cells. AB - Synaptotagmin VII (Syt VII), which has a higher Ca(2+) affinity and slower disassembly kinetics with lipid than Syt I and Syt IX, was regarded as being uninvolved in synaptic vesicle (SV) exocytosis but instead possibly as a calcium sensor for the slower kinetic phase of dense core vesicles (DCVs) release. By using high temporal resolution capacitance and amperometry measurements, it was demonstrated that the knockdown of endogenous Syt VII attenuated the fusion of DCV with the plasma membrane, reduced the amplitude of the exocytotic burst of the Ca(2+)-triggered DCV release without affecting the slope of the sustained component, and blocked the fusion pore expansion. This suggests that Syt VII is the Ca(2+) sensor of DCV fusion machinery and is an essential factor for the establishment and maintenance of the pool size of releasable DCVs in PC12 cells. PMID- 20016974 TI - Phylogeographic analysis of human papillomavirus 58. AB - Human papillomavirus 58 (HPV58) is one type of HPV with high risk of causing cervical cancer. Unusually high prevalence of HPV58 has been reported in Asia, Africa and some other areas. However, due to the scattered distribution of global data, in addition to the lack of data of some HPV58 high-incidence nations and regions, like Mainland China, a comprehensive analysis of the global geographical distribution of HPV58 remains blank so far. In this study, HPV58 from the human cervical cancer tissue was detected in Mainland China, and 14 new HPV58-E6/L1 gene sequences were obtained. Moreover, phylogeographic analysis has been conducted combining the HPV58 sequences that have been deposited in GenBank since 1985. The study result shows that the sequences detected from the Shanghai, Jiangsu and Sichuan areas are homologous with those found in the past from Hong Kong and Xi'an, China, as well as Japan and other Southeast Asian areas. Furthermore, Western Africa is considered to be the "root" source of the HPV58 variant, while Mainland China and Southeast Asia are "transit points" and the new sources of HPV58 after receiving the isolates from the "root" source; like HPV16 and HPV18, the HPV58 might also be one of the major HPV types associated with the development and spread of cervical cancer. PMID- 20016975 TI - Global transcriptional profiles of Trichophyton rubrum in response to Flucytosine. AB - Trichophyton rubrum (T.rubrum) is one of the most common human fungal pathogens that cause chronic infections of the skin and nails. To identify antifungal responsive genes, cDNA microarray analysis was performed for T. rubrum to reveal global transcriptional profiles of drug-specific responses to 5-Flucytosine (5 FC). cDNA microarray was constructed from the T. rubrum expressed sequence tag (ESTs) database, the minimum inhibitory concentration (MIC) of 5-FC was determined, and microarray hybridization and data analysis were applied. The expression pattern of 7 genes observed by microarray was confirmed by the quantitative real-time reverser transcription polymerase chain reaction (RT-PCR). Data analysis indicated that a total of 474 genes were found differentially expressed, 196 showed an increase in expression and 278 showed a decrease in expression. Marked down-regulation of genes involved in nucleotide metabolism (such as CDC21), transcription (such as E2F1), and RNA processing (such as SGN1, RIM4 and NOP1) was observed. Other genes involved in signal transduction, chaperones, inorganic ion transport, secondary metabolite biosynthesis, amino acid transport, lipid transport and potential drug resistance mechanism were also affected by 5-FC. Quantitative real-time RT-PCR of the selected genes confirmed the reliability of the microarray results. This is the first analysis of transcriptional profiles in response to 5-FC for T. rubrum. The findings may be valuable for the identification of genes involved in mechanisms of action and mechanisms of antifungal drug resistance of 5-FC. PMID- 20016976 TI - The three-dimensional structure of Infectious flacherie virus capsid determined by cryo-electron microscopy. AB - Cryo-electron microscopy and image reconstruction were used to determine the three-dimensional structure of Infectious flacherie virus (IFV). 5047 particles were selected for the final reconstruction. The FSC curve showed that the resolution of this capsid structure was 18 A. The structure is a psuedo T=3 (P=3) icosahedral capsid with a diameter of 302.4 A and a single shell thickness of 15 A. The density map showed that IFV has a smooth surface without any prominent protrude or depression. Comparison of the IFV structure with those of the insect picorna-like virus-Cricket paralysis virus (CrPV)and human picornavirus-Human rhinovirus 14 (HRV 14) revealed that the IFV structure resembles the CrPV structure. The "Rossmann canyon" is absent in both IFV and CrPV particles. The polypeptide topology of IFV VP2, IFV VP3 was predicted and the subunit location at the capsid surface was further analyzed. PMID- 20016977 TI - Expression of the difference between the Cold (Han) and Hot (Re) natures of traditional Chinese medicines (Strobal and Rhubarb) based on the cold/hot plate differentiating assay. AB - In this study, objective differences between the Cold (Han) and Hot (Re) nature of traditional Chinese medicines, e.g. Strobal and Rhubarb, are determined by using a cold/hot plate differentiation technology. A novel, self-designed cold/hot plate differentiating instrument, with methodological study, was used to investigate the intervention of Strobal and Rhubarb on the temperature tropism of mice. Compared with the ICR and BALB/c mice, it was found that KM mice on the cold/hot plate were more sensitive to the change of temperature, within the tolerant temperature range of 15-40 degrees C. The temperature tropism behavior of mice is influenced by treatment with Rhubarb and Strobal, as is the activity of ATPase in liver tissue. These trends are consistent with the definition of the Cold/Hot nature of Chinese medicines based on traditional Chinese medicinal theory. This study showed that the differences of the Cold/Hot nature of traditional Chinese medicines. might be objectively represented by the temperature tropism of animal by means of cold/hot differentiating assay. PMID- 20016978 TI - Configural processing of different topologically structured figures: an ERP study. AB - According to Chen's theory, topological differences are perceived faster than feature differences in early visual perception. We hypothesized that topological perception is caused by the sensitivity in discriminating figures with and without "holes". An ERP experiment was conducted utilizing a passive paradigm to investigate the differences in perceiving figures with "hole" and with "no-hole". The results showed differences in N170 components between figures with "holes" and with "no-holes". The inversion of the "hole" could influence the latency of N170, but the inversion of the "no-hole" could not, which indicated that global features are processed first in the "hole" perception whilst local features are given priority to the "no-hole" perception. This result was similar to studies concerning face and non-face objects, suggesting a configural processing of the "hole". PMID- 20016980 TI - Soil functional diversity analysis of a bauxite-mined restoration chronosequence. AB - Soil microorganisms are sensitive to environmental perturbations such that changes in microbial community structure and function can provide early signs of anthropogenic disturbances and even predict restoration success. We evaluated the bacterial functional diversity of un-mined and three chronosequence sites at various stages of rehabilitation (0, 10, and 20 years old) located in the Mocho Mountains of Jamaica. Samples were collected during the dry and wet seasons and analyzed for metal concentrations, microbial biomass carbon, bacterial numbers, and functional responses of soil microbiota using community-level physiological profile (CLPP) assays. Regardless of the season, un-mined soils consisted of higher microbial biomass and numbers than any of the rehabilitated sites. Additionally, the number and rate of substrates utilized and substrate evenness (the distribution of color development between the substrates) were significantly greater in the un-mined soils with carbohydrates being preferentially utilized than amino acids, polymers, carboxylic acids, and esters. To some extent, functional responses varied with the seasons but the least physiological activity was shown by the site rehabilitated in 1987 indicating long-term perturbation to this ecosystem. Small subunit ribosomal DNA (SSUrDNA)-denaturing gradient-gel electrophoresis analyses on the microbiota collected from the most preferred CLPP substrates followed by taxonomic analyses showed Proteobacteria, specifically the gamma-proteobacteria, as the most functionally active phyla, indicating a propensity of this phyla to out-compete other groups under the prevailing conditions. Additionally, multivariate statistical analyses, Shannon's diversity, and evenness indices, principal component analysis, biplot and un-weighted-pair group method with arithmetic averages dendrograms further confirmed that un-mined sites were distinctly different from the rehabilitated soils. PMID- 20016979 TI - Regulation of pH During Amelogenesis. AB - During amelogenesis, extracellular matrix proteins interact with growing hydroxyapatite crystals to create one of the most architecturally complex biological tissues. The process of enamel formation is a unique biomineralizing system characterized first by an increase in crystallite length during the secretory phase of amelogenesis, followed by a vast increase in crystallite width and thickness in the later maturation phase when organic complexes are enzymatically removed. Crystal growth is modulated by changes in the pH of the enamel microenvironment that is critical for proper enamel biomineralization. Whereas the genetic bases for most abnormal enamel phenotypes (amelogenesis imperfecta) are generally associated with mutations to enamel matrix specific genes, mutations to genes involved in pH regulation may result in severely affected enamel structure, highlighting the importance of pH regulation for normal enamel development. This review summarizes the intra- and extracellular mechanisms employed by the enamel-forming cells, ameloblasts, to maintain pH homeostasis and, also, discusses the enamel phenotypes associated with disruptions to genes involved in pH regulation. PMID- 20016981 TI - Estimation by PLFA of microbial community structure associated with the rhizosphere of Lygeum spartum and Piptatherum miliaceum growing in semiarid mine tailings. AB - The objective of this study was to compare the microbial community composition and biomass associated with the rhizosphere of a perennial gramineous species (Lygeum spartum L.) with that of an annual (Piptatherum miliaceum L.), both growing in semiarid mine tailings. We also established their relationship with the contents of potentially toxic metals as well as with indicators of soil quality. The total phospholipid fatty acid (PLFA) amount was significantly higher in the rhizosphere soil of the annual species than in the rhizosphere soil of the perennial species. The fungal/bacterial PLFA ratio was significantly greater in the perennial species compared to the annual species. The fatty acid 16:1omega5c, the fungal/bacterial PLFA ratio and monounsaturated/saturated PLFA ratio were correlated negatively with the soluble contents of toxic metals. The cyc/prec (cy17:0 + cy19:0/16:1omega7 + 18:1omega7) ratio was correlated positively with the soluble contents of Pb, Zn, Al, Ni, Cd, and Cu. The results of the PLFA analysis for profiling microbial communities and their stress status of both the plant species indicate that perennial and annual gramineous species appear equally suitable for use in programmes of revegetation of semiarid mine tailings. PMID- 20016982 TI - Implementing municipal tree planting: Los Angeles million-tree initiative. AB - Urban forests are increasingly being seen as an important infrastructure that can help cities remediate their environmental impacts. This work reports on the first steps in implementing a million tree program in Los Angeles and the ways such a biogenic-living-infrastructure has been approached. Numbers of studies have been done to quantify the benefits of urban forests, but little has been written on the process of implementing urban tree planting programs. The investigative methods were primarily qualitative, involving interviews, attending meetings and conducting literature reviews. Results indicate that multiple nonprofit and city agency programs are involved in planting and maintaining trees and this has required coordination among groups that here-to-fore were unaccustomed to having to collaborate. The main finding that emerge from this research is that the implementation of such a program in Los Angeles is more complicated than it may seem due to several interacting factors: the need to rely on multiple public and private organizations to put trees into the ground and to maintain them; coordination of these multiple efforts must be centralized, but requires a great deal of time and effort and maybe resisted by some of the partners; funding for planting and long term maintenance must be pieced together from multiple sources; acceptance of trees by residents varies by neighborhood as does tree canopy cover; appropriate nursery supply can be limited; the location of the program within the city administration is determined by who initiates the program. PMID- 20016983 TI - Remotely-sensed active fire data for protected area management: eight-year patterns in the Manas National Park, India. AB - The Terai-Duar savanna and grasslands, which once extended along most of the Himalayan foothills, now only remain in a number of protected areas. Within these localities, grassland burning is a major issue, but data on frequency and distribution of fires are limited. Here, we analysed the incidence of active fires, which only occur during the dry season (Nov.-Mar.), within a significant area of Terai grasslands: the Manas National Park (MNP), India. We obtained locations of 781 fires during the 2000-2008 dry seasons, from the Fire Information for Resource Management System (FIRMS) that delivers global MODIS hotspot/fire locations using remote sensing and GIS technologies. Annual number of fires rose significantly from around 20 at the start of the study period to over 90 after 2002, with most (85%) detected between December and January. Over half of the fires occurred in tall grasslands, but fire density was highest in wetland and riverine vegetation, dry at the time. Most burning took place near rivers, roads and the park boundary, suggesting anthropogenic origins. A kernel density map of all recorded fires indicated three heavily burnt areas in the MNP, all within the tall grasslands. Our study demonstrates, despite some technical caveats linked to fire detection technology, which is improving, that remote fire data can be a practical tool in understanding fire concentration and burning temporal patterns in highly vulnerable habitats, useful in guiding management. PMID- 20016984 TI - A framework for exploring integrated learning systems for the governance and management of public protected areas. AB - This article discusses how the concept of integrated learning systems provides a useful means of exploring the functional linkages between the governance and management of public protected areas. It presents a conceptual framework of an integrated learning system that explicitly incorporates learning processes in governance and management subsystems. The framework is premised on the assumption that an understanding of an integrated learning system is essential if we are to successfully promote learning across multiple scales as a fundamental component of adaptability in the governance and management of protected areas. The framework is used to illustrate real-world situations that reflect the nature and substance of the linkages between governance and management. Drawing on lessons from North America and Africa, the article demonstrates that the establishment and maintenance of an integrated learning system take place in a complex context which links elements of governance learning and management learning subsystems. The degree to which the two subsystems are coupled influences the performance of an integrated learning system and ultimately adaptability. Such performance is largely determined by how integrated learning processes allow for the systematic testing of societal assumptions (beliefs, values, and public interest) to enable society and protected area agencies to adapt and learn in the face of social and ecological change. It is argued that an integrated perspective provides a potentially useful framework for explaining and improving shared understanding around which the concept of adaptability is structured and implemented. PMID- 20016985 TI - Evaluating the suitability of management strategies of pure Norway spruce forests in the Black Forest area of southwest Germany for adaptation to or mitigation of climate change. AB - The study deals with the problem of evaluating management strategies for pure stands of Norway spruce (Picea abies Karst) to balance adaptation to and mitigation of climate change, taking into account multiple objectives of a forest owner. A simulation and optimization approach was used to evaluate the management of a 1000 ha model Age-Class forest, representing the age-class distribution of an area of 66,000 ha of pure Norway spruce forests in the Black Forest region of Southwest Germany. Eight silvicultural scenarios comprising five forest conversion schemes which were interpreted as "adaptation" strategies which aims at increasing the proportion of Beech, that is expected to better cope with climate change than the existing Norway spruce, and three conventional strategies including a "Do-nothing" alternative classified as "mitigation", trying to keep rather higher levels of growing stock of spruce, were simulated using the empirical growth simulator BWINPro-S. A linear programming approach was adapted to simultaneously maximize the net present values of carbon sequestration and timber production subject to the two constraints of wood even flow and partial protection of the oldest (nature protection). The optimized plan, with the global utility of 11,687 /ha in forty years, allocated a combination of silvicultural scenarios to the entire forest area. Overall, strategies classified as "mitigation" were favored, while strategies falling into the "adaptation" category were limited to the youngest age-classes in the optimal solution. Carbon sequestration of the "Do-nothing" alternative was between 1.72 and 1.85 million tons higher than the other alternatives for the entire forest area while the differences between the adaptation and mitigation approaches were approximately 133,000 tons. Sensitivity analysis showed that a carbon price of 21 /t is the threshold at which carbon sequestration is promoted, while an interest rate of above 2% would decrease the amount of carbon. PMID- 20016986 TI - Social capital in an outdoor recreation context. AB - This study examined social capital development in three all-terrain vehicles (ATV) clubs in Maine using an adapted version of Lin's (2001) social capital theory model. The structural components of social capital identified included collective assets and individual assets in the form of normative behavior and trust relationships. Also identified were counter-norms for individual ATV riders identified as having divergent norms from club members. The second component of social capital is access to and mobilization of network contacts and resources. Access networks in the context of the ATV clubs studied were identified as community and landowner relations while mobilization of resources was existent in club membership attempts toward self-governance and efforts of the statewide "umbrella" organization. Instrumental outcomes benefit society and expressive outcomes benefit the individual. Both types of returns are present in the data suggesting that ATV clubs are creating social capital. This is important information to clubs who desire to market themselves, improve their reputations, and enhance their volunteer association. It is of further interest to state governments who fund clubs through trail grants as proof that a return on investment is being realized. Theoretical and applied implications for these and other types of recreation-based volunteer associations (e.g., clubs, friends groups, advocacy groups) are presented. PMID- 20016987 TI - Behcet's disease associated with malignancy in Korea: a single center experience. AB - The aim of this study was to determine the clinical characteristics of Behcet's disease (BD) complicated with malignancy in Korea. Of 1,769 patients with BD in our hospital, 32 patients (1.8%, 21 in solid cancer, 11 in hematologic malignancy) developed cancer. In 10 of the 32 subjects (31.3%), malignancy was diagnosed before or concomitantly with BD. Twelve cases (37.5%) occurred within the first 2 years of disease and 9 cases (28.1%) occurred 5 years after the diagnosis of BD. Myelodysplastic syndrome (MDS) was the most common disease (n = 7) followed by thyroid cancer (n = 4), breast cancer, cervix cancer, stomach cancer, rectal cancer, hepatoma, aplastic anemia (n = 3, each), renal cell cancer, endometrial cancer, lymphoma (n = 1, each). There were no significant differences in the clinical characteristic between patients with or without malignancy. Intestinal involvement were more frequent in patients with malignancy than those without, but was not statistically significant (p = 0.083). Our results demonstrate that MDS and thyroid cancer are the most common hematologic disease and solid cancer associated with BD, respectively. Further studies will be required to ascertain the pathogenic link between BD and malignancy and the prevalence of malignancy in BD. PMID- 20016988 TI - Dual energy computed tomography for quantification of tissue urate deposits in tophaceous gout: help from modern physics in the management of an ancient disease. AB - Gout has been recognized for centuries but is also a modern day scourge. It is the most common type of inflammatory arthritis in men and appears to be increasing in both incidence and prevalence (Arromdee et al. in J Rheumatol 29(11):2403-2406, 2002). Despite these facts, few advances have been made in the diagnosis and treatment of gout for over 50 years. Difficult cases of gout challenge available therapeutic options. It is only recently that the Food and Drug Administration has approved febuxostat as a treatment option for patients intolerant of allopurinol. We describe a difficult case of tophaceous gout notable for several reasons: utilization of rasburicase as uricolytic treatment to dramatically reduce tissue urate burden; treatment of gout flares with interleukin-1beta inhibition; and quantification of tissue urate with novel dual energy computed tomography technology before and after uricolytic therapy. PMID- 20016989 TI - A noncontact footpad thickness assay to evaluate rheumatoid disease. AB - Measuring soft tissue thickness is an important step in rheumatoid disease research. The severity of mouse footpad swelling can be used as an indicator of disease progression. A noncontact footpad thickness assay, simplified geometry measurement system (SGMS), was developed that was able to reduce both intra- and interobserver variances during measurements. Three materials with five objects each were used in this study: hard blocks, soft sponges and mouse footpads. Thicknesses were measured using calipers or the SGMS. In the measurement of the hard block, there was no difference in measurement errors between calipers and SGMS. For the mouse footpad thickness, there was significant difference in intraobserver variances among three observers and a significant difference of interobserver variances between calipers and SGMS. In conclusions, this noncontact assay is reliable and highly reproducible for the assessment of inflammatory reactions when results are expressed as a gradual increase in footpad thickness. PMID- 20016990 TI - Nicotinic receptor channelopathies and epilepsy. AB - Characterized by sudden episodes called seizures, epilepsy was recognized long ago as a neurological disorder that can have multiple forms ranging from benign to life threatening depending upon its severity. Although several evidences indicated that genes play an important role in at least half of the patients, it is only with the advances in molecular biology and genetics that the puzzle about oligogenic and monogenic epilepsies slowly starts to unfold. The finding of an association between a monogenic form of epilepsy and a mutation in the gene encoding the neuronal nicotinic acetylcholine receptor subunit CHRNA4 marked, in 1995, a turning point in our understanding of epilepsy. It also marked the first step towards the today widely acknowledged concept of epilepsies as channelopathies. Several mutations in nicotinic acetylcholine receptor genes have, since then, been identified, and the functional properties of these mutated receptors were characterized. In this work, we review, in the light of the latest discoveries, the effects caused by the mutations on the physiological properties of the receptors and the impact of such mutations on neuronal network functions. PMID- 20016991 TI - Differential expression of CHS7 and CHS8 genes in soybean. AB - Chalcone synthase (CHS) catalyzes the first reaction specific for flavonoid and isoflavonoid biosynthesis. The soybean genome consists of nine copies of CHS genes (CHS1-CHS9) and a duplicate copy of CHS1. Even though the soybean CHS gene family members share a high degree of sequence similarity, they play different roles during plant development or in response to environmental stimuli. Our previous work on the comparison of a global gene expression in two soybean cultivars that differ in the level of total isoflavonoid accumulation has denoted the involvement of CHS7 and CHS8 genes in isoflavonoid synthesis. We have extended our effort to understand expression patterns of these two genes in soybean and in transgenic Arabidopsis. Promoter regions of CHS7 and CHS8 genes were isolated and in silico analysis performed to investigate potential transcription factor binding sites (TFBSs). The TFBSs were verified by DNase I footprint analysis. Some unique and several common TFBSs were identified in CHS7 and CHS8 promoters. We cloned beta-glucuronidase (GUS) under CHS7 and CHS8 promoters and monitored the tissue-specific GUS expression in transformed Arabidopsis. Differential GUS activity was observed in young leaves, roots, and mature pod walls of transgenic CHS7 promoter-GUS and CHS8 promoter-GUS plants. The tissue-specific expression patterns of CHS7 and CHS8 genes were determined in soybean by quantitative RT-PCR. Both CHS7 and CHS8 genes were expressed at higher levels in roots; however, overall expression pattern of these genes varied in different tissues. The results suggest that the structural diversity within CHS7 and CHS8 promoters may lead into differential activation of these genes by different inducers as well as developmental stage- and tissue-specific differences in gene expression. PMID- 20016992 TI - Rikkunshito, a traditional Japanese medicine, may relieve abdominal symptoms in rats with experimental esophagitis by improving the barrier function of epithelial cells in esophageal mucosa. AB - BACKGROUND: A traditional Japanese medicine, rikkunshito, has been reported to relieve dyspepsia symptoms. We investigated the effect of rikkunshito on RE induced abdominal dyspepsia, and performed experiments to elucidate the mechanism of that effect. METHODS: RE model rats were prepared using 8-week-old male Wistar rats, and rikkunshito was administered in drinking water. Voluntary movement was used as an index of RE-induced abdominal dyspepsia, which was monitored by an infrared sensor. On the tenth day after surgery, the total area of esophageal erosion was measured, and samples of nonerosive mucosa were collected. Using those samples, intercellular spaces of epithelial mucosa were examined by transmission electron microscopy, and the NP-40-soluble and -insoluble levels of the tight junction proteins claudin-1, -3 and -4 and their mRNAs were determined. RESULTS: Rikkunshito did not reduce the average total area of erosive lesions in the esophageal mucosa of RE model rats. On day 10, voluntary movement was significantly decreased in the RE model rats and rikkunshito significantly increased it. Nonerosive esophageal mucosa from RE rats showed dilation of intercellular spaces in epithelium, and significantly decreased claudin-3 mRNA and protein levels. Rikkunshito significantly suppressed intercellular space dilation and significantly increased the level of NP-40-insoluble claudin-3, but it did not affect the mRNA level, suggesting that it promoted tight junction formation by facilitating the translocation of proteins. CONCLUSION: Rikkunshito increased voluntary movement in RE model rats. This may have been because rikkunshito ameliorated the symptoms of RE by improving the barrier function of esophageal mucosa. PMID- 20016993 TI - CLE peptide signaling during plant development. AB - Peptide signaling in plants is a rapid developing area of research which focuses on so called peptide hormones. These signaling molecules are utilized for inter cellular communication in different developmental processes, beside the usage of the more well-known phytohormones. Probably the best studied peptide ligands in plants are the CLAVATA3 (CLV3)/ENDOSPERM SURROUNDING REGION (ESR)-related (CLE) proteins. This family of signaling polypeptides is comprised of 32 members in Arabidopsis and, with the exception of the presence of related proteins in some parasitic worms, is restricted to the plant kingdom. CLV3 is one of the founding CLE genes and is involved in stem cell niche maintenance in apical meristems during plant development. While the CLV signaling pathway is well characterized with the identification of three receptors and a stem-cell-promoting transcription factor as target, the functioning of other family members is not or poorly understood. The recent discoveries of a new type of receptor involved in CLV signaling and a functional pathway for CLE40 in root development mark the rapid progress that is made in the area of CLE peptide signaling. This review gives an overview how CLE peptides are used as signaling molecules, and how they are involved in cell-to-cell communication in concert with different known and unknown receptors in a range of developmental processes during plant development. PMID- 20016994 TI - A newly described vancomycin-resistant ST412 Enterococcus faecium predominant in Greek hospitals. AB - A total of 359 vancomycin-resistant enterococci (344 Enterococcus faecium and 15 E. faecalis) collected during 2007 from eight tertiary-care hospitals in Greece were analysed for genotypic characteristics. Four common clones, ST412, ST203, ST16 and ST17, were identified among E. faecium and one clone, ST28, among E. faecalis strains. PMID- 20016995 TI - Catheter-related Corynebacterium bacteremia: should the catheter be removed and vancomycin administered? AB - The purpose of this study was to determine the need for central venous catheter removal in patients with corynebacterial catheter-related bloodstream infections and the impact of central venous catheter retention on response to systemic antibiotic therapy and relapse. We searched the microbiology laboratory database and patients' medical records at our institution between January 2000 and December 2006. We identified 98 patients with corynebacteria infection. Most of the episodes (94%) were catheter-related. Removing the catheter did not affect the outcome of treatment, particularly when an active non-glycopeptide antibiotic was used. All Corynebacterium species isolates were susceptible to vancomycin, 54/55 (98%) to linezolid, 80/95 (84%) to rifampin, and 69/85 (81%) to tetracycline. The median duration of antibiotic therapy was 12 days (range, 0 28), and vancomycin was the most commonly used antibiotic (64%). There was a trend toward earlier fever resolution in patients treated with non-glycopeptide antibiotics compared to vancomycin, particularly if the catheter was not removed. Central venous catheter removal might not be necessary in patients with corynebacterial catheter related bloodstream infection, particularly if systemic therapy consists of non-glycopeptide antibiotics. Treatment with a systemic active antibiotic over a 7-day period appears to be adequate for resolution of the infection. PMID- 20016996 TI - A rhodopsin-like protein in Cyanophora paradoxa: gene sequence and protein immunolocalization. AB - Here, we report the DNA sequence of the rhodopsin gene in the alga Cyanophora paradoxa (Glaucophyta). The primers were designed according to the conserved regions of prokaryotic and eukaryotic rhodopsin-like proteins deposited in the GenBank. The sequence consists of 1,272 bp comprised of 5 introns. The correspondent protein, named Cyanophopsin, showed high identity to rhodopsin-like proteins of Archea, Bacteria, Fungi, and Algae. At the N-terminal, the protein is characterized by a region with no transmembrane alpha-helices (80 aa), followed by a region with 7alpha-helices (219 aa) and a shorter 35-aa C-terminal region. The DNA sequence of the N-terminal region was expressed in E. coli and the recombinant purified peptide was used as antigen in hens to obtain polyclonal antibodies. Indirect immunofluorescence in C. paradoxa cells showed a marked labeling of the muroplast (aka cyanelle) membrane. PMID- 20016998 TI - Genome-wide association studies of bladder cancer risk: a field synopsis of progress and potential applications. AB - The advent of the genome era after the completion of the Human Genome Project has resulted in intensive efforts to identify all genetic variants that modify human health and disease, including cancer. The development of genome-wide association study (GWAS) approach has facilitated this goal by unbiased examination of the entire human genome for disease association. Here, we review some of the GWAS data, particularly for bladder cancer, and assess their significance in risk prediction and prognosis. A mechanistic understanding of the risk association through functional studies and phenotypic assays is also discussed. The ultimate goal is the development of a comprehensive risk prediction model which integrates genetic, environment, and person risk factors to benefit disease diagnosis, prevention, and treatment. PMID- 20016997 TI - Platelets and innate immunity. AB - Although platelets are best known as primary mediators of hemostasis, this function intimately associates them with inflammatory processes, and it has been increasingly recognized that platelets play an active role in both innate and adaptive immunity. For example, platelet adhesive interactions with leukocytes and endothelial cells via P-selectin can lead to several pro-inflammatory events, including leukocyte rolling and activation, production of cytokine cascades, and recruitment of the leukocytes to sites of tissue damage. Superimposed on this, platelets express immunologically-related molecules such as CD40L and Toll-like receptors that have been shown to functionally modulate innate immunity. Furthermore, platelets themselves can interact with microorganisms, and several viruses have been shown to cross-react immunologically with platelet antigens. This review discusses the central role that platelets play in inflammation, linking them with varied pathological conditions, such as atherosclerosis, sepsis, and immune thrombocytopenic purpura, and suggests that platelets also act as primary mediators of our innate defences. PMID- 20016999 TI - A method of detecting carbonyl compounds in tree leaves in China. AB - BACKGROUND, AIM, AND SCOPE: Carbonyl compounds have been paid more and more attention because some carbonyl species have been proven to be carcinogenic or a risk for human health. Plant leaves are both an important emission source and an important sink of carbonyl compounds. But the research on carbonyl compounds from plant leaves is very scarce. In order to make an approach to the emission mechanism of plant leaves, a new method was established to extract carbonyl compounds from fresh plant leaves. MATERIALS, METHODS, AND RESULTS: The procedure combining derivatization with ultrasonication was developed for the fast extraction of carbonyl compounds from tree leaves. Fresh leaves (< 0.01 g) were minced and ultrasonicated in acidic 2,4-dinitrophenylhydrazine (DNPH) acetonitrile solution for 30 min and then holding 30 min to allow aldehydes and ketones in leaves to react completely with DNPH. CONCLUSIONS: The extraction process was performed under room temperature and only took 60 min. The advantages of this method were very little sample preparation, requiring short treatment time and usual equipment. Four greening trees, i.e., camphor tree (Cinnamomum camphora), sweet olive (Osmanthus fragrans), cedar (Cedrus deodara), and dawn redwood (Metasequoia glyptostroboides), were selected and extracted by this method. Seven carbonyl compounds, including formaldehyde, acetaldehyde, acetone, acrolein, p-tolualdehyde, m/o-tolualdehyde, and hexaldehyde were determined and quantified. The most common carbonyl species of the four tree leaves were formaldehyde, acrolein, and m/o-tolualdehyde. They accounted for 67.3% in cedar, 50.8% in sweet olive, 45.8% in dawn redwood, and 44.6% in camphor tree, respectively. Camphor tree had the highest leaf level of m/o-tolualdehyde with 15.0 +/- 3.4 microg g(-1)(fresh leaf weight), which indicated that camphor tree may be a bioindicator of the level of tolualdehyde or xylene in the atmosphere. By analyzing carbonyl compounds from different tree leaves, it is not only helpful for further studying the relationship between sink and emission of carbonyls from plants, but also helpful for exploring optimum plant population in urban greening. PMID- 20017000 TI - Occurrence and sources of selected phenolic endocrine disruptors in Ria de Aveiro, Portugal. AB - BACKGROUND, AIM AND SCOPE: Ria de Aveiro (Portugal) is a shallow coastal lagoon of high economic and ecological importance. Hardly any data on its chemical pollution by polar organic pollutants are available in literature. This study focused on the presence and sources of a series of phenolic endocrine-disrupting compounds (EDCs) in this area, including parabens, alkylphenolic compounds and bisphenol-A (BPA). A number of possible sources of pollution are present in the area, including the large harbours present in the lagoon, the city of Aveiro and the rivers discharging into the area. A recently constructed submarine wastewater outfall, located a few kilometres from the lagoon inlet has also been suggested as a possible source of pollution to Ria de Aveiro in several publications. The aim of the current field study was to investigate the occurrence and main sources of phenolic endocrine disruptors in Ria de Aveiro. MATERIALS AND METHODS: An extensive sampling campaign was performed, with surface water and wastewater grab samples taken at over 50 locations, in duplicate on different days. Samples were treated using solid phase extraction and analysed by liquid chromatography tandem mass spectrometry. RESULTS AND DISCUSSION: Concentrations in lagoon water were generally low: not exceeding 20 ng/L for most analytes. Levels in river water exceeded those in the lagoon by a factor 3 to 500 (o-phenylphenol (PhP) and nonylphenoxy ethoxy acetic acids (A9PEC), respectively), with concentrations up to 700 ng/L for BPA and 7,300 ng/L for A9PEC. Samples from the harbours showed EDC levels similar to those in the rest of the lagoon, but in the city of Aveiro, elevated concentrations were observed for alkylphenol ethoxylates (A9PEO), A9PEC, PhP and BPA. Wastewater effluents showed low levels for parabens, whilst alkylphenolic compounds reached several micrograms per litre. The effluents are discharged into the ocean via a submarine outfall, but as marine water near the outfall showed slightly elevated concentrations only for A9PEO, it does not seem to be a significant source of these EDCs for the area. CONCLUSIONS: All the studied phenolic EDCs were detected in the study area, with high levels found in some of the rivers discharging into the lagoon, and generally low concentrations in the lagoon itself. The main sources for all investigated EDCs were the rivers Caster and Antua which discharge into the lagoon. The city of Aveiro was identified as a secondary source. As the tidal water exchange volume is much larger than the freshwater input from the rivers, concentrations of phenolic EDCs remained low in the lagoon. PMID- 20017001 TI - Diagnostic accuracy of 64-slice computed tomography coronary angiography in a large population of patients without revascularisation: registry data and review of multicentre trials. AB - PURPOSE: This study was undertaken to evaluate the diagnostic accuracy of computed tomography coronary angiography (CT-CA) for the detection of significant coronary artery stenosis (> or =50% lumen reduction) compared with conventional coronary angiography (CCA) in a registry and to review major multicentre trials. MATERIALS AND METHODS: A total of 1,372 patients (882 men, 490 women; mean age 59.3+/-11.9 years) in sinus rhythm were studied with CT-CA (64-slice technology) and CCA. The diagnostic accuracy of CT-CA was evaluated against quantitative CCA as a reference standard for coronary artery stenosis. Positive and negative likelihood ratios and inter- and intraobserver agreement were calculated. RESULTS: The prevalence of disease was 53%. CCA demonstrated the absence of significant coronary artery disease in 46.6% (639/1372), single-vessel disease in 24.7% (337/1372) and multivessel disease in 28.9% (396/1372) of patients. In per patient analysis sensitivity, specificity and positive and negative predictive value of CT-CA were 99% [confidence interval (CI) 97-99], 92% (CI 89-94), 94% (CI 91-95) and 99% (CI 97-99), respectively. Per-patient and per-segment likelihood ratios (LR+=12.4 and LR-=0.011; LR+=18.3 and LR-=0.064, respectively), were good. Inter- and intraobserver variability was 0.78 and 0.85, respectively. CONCLUSIONS: CT-CA is a reliable diagnostic modality both in terms of sensitivity and negative predictive value. Differences in trial results are also due to the different parameters used for patient inclusion. PMID- 20017002 TI - Dual-time-point [18F]-FDG PET/CT in the diagnostic evaluation of suspicious breast lesions. AB - PURPOSE: The authors sought to evaluate whether the reacquisition of images 3 h after administration of radiotracer improves the sensitivity of fluorine-18 fluorodeoxyglucose positron emission tomography computed tomography ([(18)F]-FDG PET/CT) in patients with suspicious breast lesions. MATERIALS AND METHODS: Forty eight patients with 59 breast lesions underwent an [(18)F]-FDG PET/CT study in the prone position with a dual-time-point acquisition performed in the early phase 1 h after FDG administration (PET-1) and in the delayed phase 3 h after FDG administration (PET-2). Both examinations were evaluated qualitatively and semiquantitatively with calculation of the mean percentage variation of the standard uptake values (Delta% SUV(max)) between PET-1 and PET-2. All lesions with an SUV(max) >or=2.5 at PET-1 or a reduction in SUV between PET-1 and PET-2 were considered benign. The definitive histopathological diagnosis was available for all patients included in the study. RESULTS: The dual-time-point acquisition of [(18)F]-FDG PET/CT displayed an accuracy of 85% for lesions with an SUV(max) >or=2.5 and/or positive Delta% SUV(max), with sensitivity and specificity values of 81% and 100% compared with 69%, 63% (both p<0.001) and 100% (p=n.s.), respectively, for the single-time-point acquisition. Malignant lesions showed an increase in FDG uptake between PET-1 and PET-2, with a Delta% SUV(max) of 10+/-7 (p<0.04). In contrast, benign lesions showed a decrease in SUV between PET-1 and PET-2, with a Delta% SUV(max) of -21+/-7 (p<0.001). CONCLUSIONS: The delayed repeat acquisition of PET images improves the accuracy of [(18)F]-FDG PET/CT in patients with suspicious breast lesions with respect to the single-time-point acquisition. In addition, malignant breast lesions displayed an increase in FDG uptake over time, whereas benign lesions showed a reduction. These variations in FDG uptake between PET-1 and PET-2 are a reliable parameter that can be used for differentiating between benign and malignant breast lesions. PMID- 20017003 TI - Role of 64-slice cardiac computed tomography in the evaluation of patients with non-ST-elevation acute coronary syndrome. AB - PURPOSE: This study was done to evaluate the feasibility, sensitivity and specificity of 64-slice computed tomography (CT) in identifying haemodynamically significant (>50%) coronary artery stenoses in patients with suspected acute coronary syndrome (ACS) by correlating the CT findings with the clinical event and data provided by conventional coronary angiography (CCA). MATERIALS AND METHODS: Sixty-four patients (38 men and 26 women; mean age 65 years; range+/-10 years) presenting to our hospital's emergency department with a clinical suspicion of ACS were studied with 64-slice CT followed by CCA within 24 h of arrival. RESULTS: Two patients (3.1%) were excluded from the analysis due to artefacts. Per-patient analysis in the remaining 62 patients identified 24 cases (38.7%) of negative CT findings (no stenoses or stenoses <50%), 35 cases of positive CT findings (56.4%) with identification of the culprit lesion, two cases in which the culprit lesion was not identified and one patient with unconfirmed stenosis. Sensitivity and specificity were 94.6% and 96%, respectively. Per vessel analysis (186 vessels) revealed 17 non-evaluable vessels (9.1%) due to motion artefacts, 61 vessels (32.8%) with stenosis >50%, seven overestimated vessels (3.7%) due to extensive calcifications, three vessels (1.6%) with underestimated stenosis and 98 vessels (52.6%) without stenosis. Sensitivity and specificity were 95.3% and 93.3%, respectively. CONCLUSIONS: In this type of emergency, coronary CT angiography could lead to considerably lower healthcare costs by identifying patients without coronary disease and allowing immediate discharge without any need for further diagnostic procedures. PMID- 20017004 TI - Impact of contrast material volume on quantitative assessment of reperfused acute myocardial infarction using delayed-enhancement 64-slice CT: experience in a porcine model. AB - PURPOSE: Our purpose in this study was to compare the impact of contrast material volume in delayed-enhancement computer tomography (CT) imaging for assessing acute reperfused myocardial infarction. MATERIALS AND METHODS: In five domestic pigs (20-30 kg), the circumflex coronary artery (CX) was balloon-occluded for 2 h followed by reperfusion. After 5 days, CT imaging was performed after intravenous administration of iodinated contrast material (Iomeprol 400 mgI/ml; Bracco, Italy). A 64-slice multidetector CT (MDCT) (Sensation 64, Siemens) scanner was used for imaging, with standard angiography characteristics. Three scans were performed: first, coronary angiography at first pass with 1.25 gI/kg of contrast material (ART); and remaining delayed-enhancement (DE(1)-DE(2)) 15 min after administration of 1.25 (DE(1)) and 15 min after additional administration of 2.50 gI/kg (=total 3.75 gI/kg - DE(2)). Mean heart rate decreased to 51+/-9 bpm after intravenous administration of Zatebradine (10 mg/kg). Data sets were reconstructed during the end-diastolic phase of the cardiac cycle. Areas of infarction-enhanced (DE), no-reflow (no-reflow) and remote myocardial [remote left ventricle (LV)] were manually contoured. CT attenuation values (Hounsfield units) were measured using five regions of interest: DE, no-reflow, remote LV, left ventricular cavity (lumen LV) and in air. Differences, correlations, signal to-noise ratio (SNR) and contrast-to-noise ratio (CNR) were calculated. RESULTS: We found significant differences between the attenuation of DE, no-reflow and remote LV (p<0.001). DE and no-reflow size were assessed accurately with DEMDCT. In particular, SNR and CNR showed higher values in DE(2) (approximately 6.0 and 3.5, respectively; r(2)=0.90) vs. DE(1) (approximately 4.0 and 2.2, respectively; r(2)=0.85). CONCLUSIONS: The increase of contrast material volume determines a significant improvement in myocardial infarction image quality with DE-MDCT. PMID- 20017005 TI - Computed tomography and thymoma: distinctive findings in invasive and noninvasive thymoma and predictive features of recurrence. AB - PURPOSE: Our goal was to assess the computed tomography (CT) imaging findings of thymoma and to correlate these features with Masaoka staging system and prognosis. MATERIALS AND METHODS: CT findings of thymoma were analysed in 58 patients who had undergone surgery between January 2002 and September 2007. All cases were classified according to the Masaoka staging system. The presence of various CT findings was correlated with tumour invasiveness and recurrence. In statistical analysis, a p value <0.05 was interpreted as significant. RESULTS: The study found 26 noninvasive thymomas and 32 invasive thymomas. Invasive thymomas were more likely to be greater in size (p<0.01), with lobulated or irregular contours (p<0.02), a necrotic or cystic component (p<0.04), foci of calcification (p<0.05) and heterogeneous contrast enhancement (p<0.01) than were noninvasive thymomas. Disease progression developed in nine of 58 patients. Tumour recurrence and metastasis correlated with greater size (p<0.04), lobulated or irregular contours (p<0.01), complete mediastinal fat obliteration (p<0.01), great vessel invasion (p<0.01) and pleural implants (p<0.02). CONCLUSIONS: CT is useful in differentiating invasive from noninvasive thymomas and plays an important role in evaluating and treating these patients for multimodal therapy with neoadjuvant approaches. Moreover, CT findings may serve as predictors of postoperative recurrence or metastasis. PMID- 20017006 TI - Infratentorial lesion volume correlates with sensory functional system in multiple sclerosis patients: a 3.0-Tesla MRI study. AB - PURPOSE: This study sought to correlate lesion volume in infratentorial areas using 3.0-T proton-density (PD)-weighted images with disability scales and appropriate functional system scores in patients with multiple sclerosis (MS). MATERIALS AND METHODS: We examined 20 consecutive patients (13 women and 7 men) with a median age of 47 years (range 26-70). Neurological examination included the Expanded Disability Status Scale and its functional systems, the Barthel Index (BI) and the Rivermead Mobility Index (RMI). MRI scans were performed on a system operating at 3.0 T using a quadrature birdcage head coil. Acquired images imported as Digital Imaging and Communication in Medicine (DICOM) files, and the region of interest (ROI) files were converted to Neuroimaging Informatics Technology Initiative (NIfTI) format and normalised to the Montreal Neurological Institute (MNI) standard template. An automated segmentation algorithm was used to distinguish between supratentorial and infratentorial areas. Normalisation to the magnetisation-prepared rapid acquisition with gradient echo (MPRAGE) T1 weighted sequence allowed lesion volume estimation in the different anatomical areas. RESULTS: A significant correlation was found between infratentorial lesion volume and the sensory functional system score (rho=0.76, p=0.002). No significant correlation was found between supratentorial lesion volume and Expanded Disability Status Scale (EDSS), RMI and BI scores. CONCLUSIONS: The described method, by means of anatomical assignment of MS lesions, allows detection of significant correlation coefficients between clinical and MRI lesion burden in MS patients at the infratentorial level. PMID- 20017007 TI - Low-dose unenhanced CT protocols according to individual body size for evaluating suspected renal colic: cumulative radiation exposures. AB - PURPOSE: The aim of this study was to assess the radiation dose of dose-reduced unenhanced abdominal multidetector computed tomography (MDCT) scan protocols for suspected renal colic in patients within normal weight range and overweight-obese patients and to record the cumulative dose of repeated examinations. MATERIALS AND METHODS: Over a 2-year period, we performed 1,026 unenhanced CT examinations for urolithiasis; among these, 675 were performed on 636 patients referred from the emergency department. Patients were divided into two groups on the basis of body mass index (BMI): normal weight (BMI <25 kg/m(2) group 1); overweight and obese (BMI >25 kg/m(2) group 2). For patients in group 1 and group 2, the protocols of our 64-row scanner prescribe tube current settings at 70 mAs and 150 mAs, respectively. The dose-length product (DLP) estimated by using the manufacturer's software was converted into effective dose (ED). RESULTS: Mean DLP and ED were 177 and 345 mGy/cm and 2.4 and 4.8 mSv for group 1 and group 2, respectively. A subset of 25 patients (3.7%) underwent two or more examinations, with estimated ED ranging from 4.8 to 19.2 mSv. CONCLUSIONS: Although radiation dose is nearly double in overweight-obese patients undergoing MDCT, it remains lower than that delivered by a standard-dose protocol. Patients with flank pain, who are often young, are at increased risk for serial CT examinations. Use of a low-dose protocol is mandatory in both normal-weight and obese patients to minimise radiation exposure. PMID- 20017008 TI - Inflammatory breast cancer: MR imaging findings. AB - PURPOSE: This paper describes the magnetic resonance (MR) imaging features of primary inflammatory breast cancer (IBC). MATERIALS AND METHODS: Two radiologists reviewed the MR examinations of 14 women with a pathological diagnosis of IBC. Images were assessed for skin thickening, oedema, nipple retraction, architectural distortion, type and extent of parenchymal and cutaneous enhancement and enhancement kinetics over time, axillary and internal mammary lymphadenopathy, pectoral muscle enhancement and additional findings. RESULTS: Skin thickening was identified in eight patients (58%), oedema in nine (64%), nipple retraction in two (14%), architectural distortion in eight (58%), mass like enhancement in five (36%), non-mass-like enhancement in nine (64%) with washout enhancement curve in 12 (86%) and plateau curve in two (14%), axillary lymphadenopathy in 12 (86%) and internal mammary artery lymphadenopathy in two (14%), and pectoral muscle enhancement in one (7%). Additional findings included increased breast volume in two patients (14%), prepectoral fluid in four (28%) and hypertrophic internal mammary artery in three (21%). CONCLUSIONS: The most characteristic MR findings of IBC are skin thickening, oedema, architectural distortion, mass-like enhancement with washout curve and axillary lymphadenopathy; less frequent ones are nipple retraction, mass-like enhancement and internal mammary lymphadenopathy. Prepectoral fluid is frequent but is not a sign of infiltration. PMID- 20017009 TI - Agenesis of the corpus callosum in fetuses with mild ventriculomegaly: role of MR imaging. AB - PURPOSE: We evaluated the role of magnetic resonance (MR) imaging in the diagnosis of corpus callosum agenesis - isolated or associated with other anomalies - in fetuses with mild cerebral ventriculomegaly, as depicted at prenatal sonography. MATERIAL AND METHODS: Between January 2005 and March 2007, 33 fetuses with a mean gestational age of 28.9 weeks (range 17-37) and mild ventriculomegaly diagnosed at prenatal sonography were included in this prospective study. All fetuses underwent MR imaging according to the following protocol: half-Fourier T2-weighted images along the three orthogonal plane according to the longitudinal axis of the mother, and subsequently three orthogonal planes were acquired according to the fetal brain. Quantitative image analysis included the size of the transverse diameter of the lateral ventricles, in the axial plane, and the thickness of the adjacent cerebral cortex. Qualitative image analysis included morphology of the lateral ventricles (normal, parallel pattern colpocephaly), signal intensity changes of the fetal brain, interruption of the germinative matrix, agenesis of the corpus callosum (complete/partial) and associated malformations. Postnatal physical examination and diagnostic imaging, as well as surgery, were the standard of diagnosis. RESULTS: Mean axial diameter of the lateral ventricle was 11.6 mm (range 10-15 mm), and mean thickness of the adjacent cerebral cortex was 2.1 mm (range 1.8-3 mm); 23/33 fetuses (70%) showed normal morphology of the lateral ventricles, and 8/33 (24%) showed abnormal morphology (parallel pattern, colpocephaly). The entire corpus callosum was visualised in 20/33 fetuses (60%). In 8/33 fetuses (25%), partial agenesis was diagnosed, whereas in 5/33 (15%), there was hypogenesis. In 6/13 fetuses (46%), isolated corpus callosum agenesis was detected, and two cases of hypogenesis of the corpus callosum were misinterpreted - overestimated in one case and underestimated in another. CONCLUSIONS: MR imaging may prove to be a useful second-line imaging modality in the prenatal diagnosis of corpus callosum agenesis in fetuses with mild ventriculomegaly. PMID- 20017010 TI - Effects of two variants of Roux-en-Y Gastric bypass on metabolism behaviour: focus on plasma ghrelin concentrations over a 2-year follow-up. AB - BACKGROUND: To study the effects of two variants of Roux-en-Y gastric bypass (RYGBP) on plasma ghrelin concentrations according to different exposure of gastric fundus to the nutrient pathway. METHODS: A prospective longitudinal 2 year follow-up study was conducted. Ninety-six morbidly obese (MO) patients (age range: 41.6 +/- 9.6 years, body mass index: 53 +/- 9.5 kg/m(2)) were assigned to two bariatric surgical (BS) procedures: one that preserves food contact with gastric fundus (ringed RYGBP, n = 50) and the other that avoids it (modified RYGBP, n = 46). Different anthropometric and biochemical parameters were studied, focusing on ghrelin concentrations at baseline and 6, 12, and 24 months post-BS. RESULTS: At 24 months post-BS, all metabolic parameters studied had improved in all patients compared with those at 1-year follow-up and baseline (p < 0.05). However, high-density lipoprotein cholesterol concentrations took 2 years to normalise in 80% of patients, interleukin-6 decreased significantly in relation to baseline only after 2 years from BS (p < 0.001), and tumour necrosis factor alpha concentrations did not significantly decrease during the 2 years of follow up. Plasma ghrelin concentrations increased in both surgical groups compare to baseline during the first year (24.6% in modified RYGBP and 36.62% in ringed RYGBP) and remained stable at the second year of follow-up, with no statistical differences between groups. CONCLUSIONS: In the second year of follow-up after BS, morbidity continued to improve in MO patients despite a lesser weight loss in relation to the first year. An increase in plasma ghrelin concentrations was found, regardless of nutrient contact with gastric fundus. Furthermore, changes in plasma ghrelin concentrations appeared to be independent of weight loss. PMID- 20017011 TI - Colon delivery of budesonide: evaluation of chitosan-chondroitin sulfate interpolymer complex. AB - The present study was aimed at formulating tablets comprising of coating susceptible to microbial enzyme degradation for releasing budesonide in the colon. Tablets prepared by using Avicel pH 102 as diluent and Eudragit L100-55 as binder were coated to a weight gain of 10% w/w employing aqueous mixtures containing chitosan (CH) and chondroitin sulfate (CS). The interpolymer complex between CH and CS was characterized using Fourier transform infrared (FTIR) and differential scanning calorimetery (DSC) studies. The tablets were evaluated for release of budesonide through in vitro in vivo studies. Formation of bonds between -COO(-) and -OSO3(-) groups of CS and -NH3+ groups of CH was evident in the FTIR spectra of these interpolymer complexed (IPC) films. The DSC thermograms of these films revealed one endothermic transition between 190 degrees C and 205 degrees C, suggesting the formation of new bonds in the IPC. The pH sensitive swelling exhibited by these films was observed to be a function of CH concentration. Tablets coated with aqueous mixtures containing 40:60 or 50:50 ratio of CH/CS totally prevented the release of budesonide in pH 1.2 buffer. The peaks (FTIR) and endothermic transitions (DSC) characteristic of interpolymer complexation were observed to remain unaffected after sequential exposure of the films to pH 1.2 and pH 7.4 buffer IP. This proved the versatility of these IPC films for colon delivery. C (max) of 1,168.99 and 1,174.2 ng/mL, respectively, at 12 and 8 h post-oral dosing of tablets coated with 40:60 or 50:50 ratio of CH/CS was observed in rats. The aqueous CH/CS (40:60) coating could provide a facile method for delivering budesonide to the colon. PMID- 20017012 TI - Structural biology of NCAM. PMID- 20017013 TI - Extracellular protein interactions mediated by the neural cell adhesion molecule, NCAM: heterophilic interactions between NCAM and cell adhesion molecules, extracellular matrix proteins, and viruses. PMID- 20017014 TI - Intracelluar ligands of NCAM. PMID- 20017015 TI - NCAM and the FGF-receptor. PMID- 20017016 TI - The role of ATP in the regulation of NCAM function. PMID- 20017017 TI - Polysialylation of NCAM. PMID- 20017018 TI - Structural basis for the polysialylation of the neural cell adhesion molecule. PMID- 20017019 TI - The role of PSA-NCAM in adult neurogenesis. PMID- 20017020 TI - Use of PSA-NCAM in repair of the central nervous system. PMID- 20017021 TI - Signaling pathways involved in NCAM-induced neurite outgrowth. PMID- 20017023 TI - The neural cell adhesion molecule NCAM and lipid rafts. PMID- 20017022 TI - Role of the growth-associated protein GAP-43 in NCAM-mediated neurite outgrowth. PMID- 20017024 TI - The neural cell adhesion molecule and epidermal growth factor receptor: signaling crosstalk. PMID- 20017025 TI - Biosynthesis of NCAM. PMID- 20017026 TI - Soluble NCAM. PMID- 20017027 TI - Role of NCAM in spine dynamics and synaptogenesis. PMID- 20017028 TI - NCAM in long-term potentiation and learning. PMID- 20017029 TI - Role of NCAM in emotion and learning. PMID- 20017030 TI - NCAM in neuropsychiatric and neurodegenerative disorders. PMID- 20017031 TI - Neural cell adhesion molecule in cancer: expression and mechanisms. PMID- 20017032 TI - NCAM mimetic peptides: an update. PMID- 20017033 TI - Synthetic NCAM-derived ligands of the fibroblast growth factor receptor. PMID- 20017034 TI - Dendritic spine and synapse morphological alterations induced by a neural cell adhesion molecule mimetic. PMID- 20017035 TI - Fasciclin II: the NCAM ortholog in Drosophila melanogaster. PMID- 20017036 TI - The neural cell adhesion molecule NCAM2/OCAM/RNCAM, a close relative to NCAM. PMID- 20017037 TI - Honoring Dr. Elisabeth Bock. PMID- 20017038 TI - Abductive inference and delusional belief. AB - Delusional beliefs have sometimes been considered as rational inferences from abnormal experiences. We explore this idea in more detail, making the following points. First, the abnormalities of cognition that initially prompt the entertaining of a delusional belief are not always conscious and since we prefer to restrict the term "experience" to consciousness we refer to "abnormal data" rather than "abnormal experience". Second, we argue that in relation to many delusions (we consider seven) one can clearly identify what the abnormal cognitive data are which prompted the delusion and what the neuropsychological impairment is which is responsible for the occurrence of these data; but one can equally clearly point to cases where this impairment is present but delusion is not. So the impairment is not sufficient for delusion to occur: a second cognitive impairment, one that affects the ability to evaluate beliefs, must also be present. Third (and this is the main thrust of our paper), we consider in detail what the nature of the inference is that leads from the abnormal data to the belief. This is not deductive inference and it is not inference by enumerative induction; it is abductive inference. We offer a Bayesian account of abductive inference and apply it to the explanation of delusional belief. PMID- 20017039 TI - The misidentification syndromes as mindreading disorders. AB - The patient with Capgras' syndrome claims that people very familiar to him have been replaced by impostors. I argue that this disorder is due to the destruction of a representation that the patient has of the mind of the familiar person. This creates the appearance of a familiar body and face, but without the familiar personality, beliefs, and thoughts. The posterior site of damage in Capgras' is often reported to be the temporoparietal junction, an area that has a role in the mindreading system, a connected system of cortical areas that allow us to attribute mental states to others. Just as the Capgras' patient claims that that man is not his father, the patient with asomatognosia claims that his arm is not really his. A similar account applies here, in that a nearby brain area, the supramarginal gyrus, is damaged. This area works in concert with the temporoparietal junction and other areas to produce a large representation of a mind inside a body situated in an environment. Damage to the mind-representing part of this system (coupled with damage to executive processes in the prefrontal lobes) causes Capgras' syndrome, whereas damage to the body-representing part of this system (also coupled with executive damage) causes asomatognosia. PMID- 20017040 TI - Kaupapa Maori Action Research to improve heart disease services in Aotearoa, New Zealand. AB - Action Research can be a powerful tool for change and improvement in health services for indigenous people when utilised within an appropriate framework. The project Maori Utilisation & Experience of Ischaemic Heart Disease Management illustrates this convergence in its use of Kaupapa Maori Action Research methods in its efforts to improve the health and well-being of Maori within the northern region of Aotearoa/New Zealand. We outline the research processes and outcomes obtained through the application of 'by Maori for Maori' approaches to understanding Maori pathways and barriers to care for ischaemic heart disease. Maori understandings of their illness and experiences of treatment, and healthcare providers' perspectives on care of Maori with ischaemic heart disease, were combined into Maori-led actions to improve service provision. The paper examines critical factors in an action research approach to health service innovations and implications for efforts to reduce entrenched health disparities. PMID- 20017041 TI - Economic valuations in global health. AB - This paper deals with three issues concerning economic valuation in global health. First, we argue that the economic value of health delivery in resource limited settings is not fully captured through the adding up of successive assessments of individual interventions. Second, we suggest that economic valuations can be used to identify social barriers to the success of health technologies in resource-limited settings. Third, we briefly discuss new directions for research on economic valuations given the interdependency between poor health and economic impoverishment. PMID- 20017042 TI - Changes in substrate access did not affect early feather-pecking behavior in two strains of laying hen chicks. AB - Feather pecking, commonly found in flocks of laying hens (Gallus gallus), is detrimental to bird welfare. Thought to cause this problem is the normal housing of layers without a floor substrate. Some evidence suggests that early substrate access decreases later feather pecking. However, there has been little research on the immediate effects of a change in substrate availability on bird welfare, although environmental modifications like this are often done when brooding and rearing laying hen chicks. To investigate this, the behavior of two strains of laying hen chicks was recorded for 4 weeks. The study kept the birds on either wire or peat moss for 14 days and then switched half the chicks to the other flooring. Early feather pecking was not significantly different for birds started on peat moss and switched to wire than for birds only on wire (p > .05). Because moving chicks from peat moss to wire did not cause additional welfare problems, the study recommends that chicks be kept on a substrate when young as feather pecking levels are lower and immediate welfare is improved compared with birds kept only on wire. PMID- 20017043 TI - Characteristics of relinquished dogs and their owners at 14 rehoming centers in the United Kingdom. AB - In the United Kingdom, each year many companion animal (pet) caregivers (owners) hand over dogs to shelters for rehoming. Studies conducted in the United States and Australia have shown that accommodation issues and problematic behaviors are the most common reasons for dogs to be relinquished. The purpose of this study was to provide a clearer understanding of common characteristics of relinquished dogs in the United Kingdom. A descriptive study conducted during 2005 collected data on 2,806 dogs relinquished to Dogs Trust's rehoming centers in the United Kingdom. The most common reasons for dogs to be relinquished were their problematic behaviors and their need for more attention than the owner could provide. The results of this study identify several common characteristics of relinquished dogs as well as differences between its results and those found in studies conducted in the United States and Australia. An improved knowledge of characteristics of relinquished dogs should inform the development of strategies for improved management of the rehoming process. PMID- 20017044 TI - Behavior of a solitary sociable female bottlenose dolphin (Tursiops truncatus) off the coast of Kent, Southeast England. AB - This article provides a report of the behavior of a solitary sociable dolphin studied on the southeast coast of England in 2007. This is the first study of its kind in which behavior of such a nonhuman animal was systematically studied. By the time of this study, this young female was highly interactive with people in the water. People accompanied the dolphin for 18.4% of the 100 hr of observation, and their presence changed her behavior. The study recorded 39 different behaviors; feeding and resting behaviors declined in frequency in the presence of people. In addition, the dolphin exhibited behavior possibly hazardous to people in the water, which included preventing swimmers from leaving the water. The dolphin received several wounds, at least one of which was life-threatening. This article discusses the welfare implications for such animals. PMID- 20017045 TI - The effects of chronic exposure to common bedding materials on the metabolic rate and overall health of male CD-1 mice. AB - Anecdotes and personal Web pages claim that cedar and pine beddings cause respiratory distress in rodents, although no previous research could be found to support these claims. There have, however, been published studies of respiratory distress in cedar and pine mill workers. That research links exposure to wood dust to asthma and to bronchial and alveolar damage in humans. This study looks at the effects of 3 types of bedding (CareFRESH Original, cedar, and pine) on the growth, food intake, oxygen consumption, IgE antibody concentrations, and general appearance and behavior in male CD-1 mice. Mice who were housed on these beddings for approximately 4 months did not show significant differences in any of these variables. This suggests that these 3 materials provide equally healthy substrates for long-term rearing of mice and possibly other rodents. PMID- 20017046 TI - The effects of husbandry training on stereotypic pacing in captive African wild dogs (Lycaon pictus). AB - To examine the effects of operant conditioning on stereotypic pacing in 3 female African wild dogs located at the Franklin Park Zoo in Boston, this study made recordings of pacing behavior immediately following individual sessions of husbandry training and 2 no-training conditions. The study found significant differences in the percentage of observations spent in stereotypic pacing behaviors for all 3 dogs among the 3 different conditions. The authors discuss the data in terms of the contribution of motivated tasks to the effects and the role of food deprivation in the expression of stereotypic pacing. The study suggests that even short periods of training may improve the African wild dogs' welfare by reducing stereotypic pacing following the conditioning sessions. PMID- 20017047 TI - Emergency and disaster planning at Ohio animal shelters. AB - Results of a cross-sectional study to determine the level of emergency and disaster response planning at Ohio nonhuman animal shelters and the role Ohio agencies have in emergency and disaster response planning in their communities indicated a lack of preparedness coupled with underutilization of the agencies as a resource. A total of 115 agencies (68%) responded to a standardized survey mailed to 170 Ohio agencies. Most (68%) agencies agreed that emergency and disaster response planning was important to their organization, although only 13% of agencies had completed a written emergency and disaster response plan. The majority (80%) of agencies indicated they would provide critical resources in an emergency or disaster in their community. Only 38 (33%) of the responding agencies were aware of the PETS Act of 2006. Although many agencies indicated the importance of an emergency and disaster plan, there may be insufficient resources, including time and proper training, available to ensure plans are developed. Improved coordination among veterinarians, local veterinary medical associations, emergency preparedness agencies, and animal shelters would enhance the relief efforts in a crisis. PMID- 20017048 TI - Laboratory rodent welfare: thinking outside the cage. AB - This commentary presents the case against housing rats and mice in laboratory cages; the commentary bases its case on their sentience, natural history, and the varied detriments of laboratory conditions. The commentary gives 5 arguments to support this position: (a) rats and mice have a high degree of sentience and can suffer, (b) laboratory environments cause suffering, (c) rats and mice in the wild have discrete behavioral needs, (d) rats and mice bred for many generations in the laboratory retain these needs, and (e) these needs are not met in laboratory cages. PMID- 20017049 TI - Animal welfare beyond the cage...and beyond the evidence? AB - In "Laboratory Rodent Welfare: Thinking Outside the Cage," Balcombe (2010/this issue) suggests that laboratory cage housing is damaging to rats and mice because it does not meet their evolved needs and may damage their psychological and physical health. The article also indicates that larger and more enriched spaces reduce aggression and mortality and improve the health and friendliness of rodents in the laboratory. Remarkably, many of the studies cited as supporting these assertions fail to provide data bearing on the issues involved or may even report findings opposite to those described by Balcombe, whereas many studies that are highly relevant to these issues are not cited or described. Moreover, although the "evolved needs" of rats and mice are presented as the basis for an analysis of rodent welfare, the important and well-documented changes in needs- or motivation-related behaviors of a rodent in the laboratory (due to human selection over hundreds of generations) is ignored. This pattern of disconnections between data and conclusions is so pervasive as to demolish the scientific value of the exposition. PMID- 20017050 TI - Thinking outside our cages. AB - Researchers seem to be stuck reiterating the now-familiar argument that barren boxes are bad for welfare and that rodents are due ethical consideration. But the prerequisites for real progress are new kinds of arguments, new types of data, and removal of very real practical and cultural obstacles to implementation of meaningful enrichment. We must discover what we have to do to effectively change the practices of people who have care and control of rodents in the laboratory, not just husbandry staff but those who develop the institution's protocols, job descriptions, and resourcing. Researchers are inventers of information, and like any inventor we should experience no satisfaction until our ideas are fully implemented-and we must be an active participant in that process. If we are asking animal caretakers to make deep, paradigmatic changes in their thinking, it is imperative that we in turn develop an emotionally positive understanding of areas important to them. For unless the welfare advocates truly understand the issues such as budgets, biosecurity, and branding, why should the people responsible for those subjects listen to us? PMID- 20017051 TI - Toward genuine rodent welfare: response to reviewer comments. PMID- 20017053 TI - Physiological monitoring in firefighter ensembles: wearable plethysmographic sensor vest versus standard equipment. AB - We evaluated the accuracy of a wearable sensor vest for real-time monitoring of physiological responses to treadmill exercise. Ten subjects in standard firefighter ensembles, treadmill exercising at 50% VO(2) max, had heart rate (HR), respiratory rate (RR), skin temperature (T(sk)), oxygen saturation (SaO(2)), tidal volume (V(T)), and minute ventilation (V(E)) recorded concurrently by a wearable plethysmographic sensor vest and standard laboratory physiological monitoring equipment for comparison. A high degree of correlation was noted for most of the measured variables [HR (r = 0.99), RR (r = 0.98), T(sk) (r = 0.98), V(E) (r = 0.88), and SaO(2) (r = 0.79)]. V(T) (r = 0.60) had a moderate correlation, although a paired differences analysis showed a mean paired difference of -0.03 L. This mean paired difference represents a 1.92% variation for V(T). Data from the wearable sensor vest is comparable to data captured from standard laboratory physiological monitoring equipment on subjects wearing standard firefighter ensembles while exercising at a moderate work rate. This study demonstrates the accuracy of the wearable sensor technology for these physiological parameters under these conditions and suggests that it could be useful for actual field studies of firefighters in traditional firefighting gear. PMID- 20017054 TI - Nanoparticle emission assessment technique (NEAT) for the identification and measurement of potential inhalation exposure to engineered nanomaterials--part A. AB - There are currently no exposure limits specific to engineered nanomaterial nor any national or international consensus standards on measurement techniques for nanomaterials in the workplace. However, facilities engaged in the production and use of engineered nanomaterials have expressed an interest in learning whether the potential for worker exposure exists. To assist with answering this question, the National Institute for Occupational Safety and Health established a nanotechnology field research team whose primary goal was to visit facilities and evaluate the potential for release of nanomaterials and worker exposure. The team identified numerous techniques to measure airborne nanomaterials with respect to particle size, mass, surface area, number concentration, and composition. However, some of these techniques lack specificity and field portability and are difficult to use and expensive when applied to routine exposure assessment. This article describes the nanoparticle emission assessment technique (NEAT) that uses a combination of measurement techniques and instruments to assess potential inhalation exposures in facilities that handle or produce engineered nanomaterials. The NEAT utilizes portable direct-reading instrumentation supplemented by a pair of filter-based air samples (source-specific and personal breathing zone). The use of the filter-based samples are crucial for identification purposes because particle counters are generally insensitive to particle source or composition and make it difficult to differentiate between incidental and process-related nanomaterials using number concentration alone. Results from using the NEAT at 12 facilities are presented in the companion article (Part B) in this issue. PMID- 20017055 TI - The impact of Ontario smoke-free legislation on secondhand smoke in enclosed public places. AB - Many studies have evaluated the impact of indoor smoking bans on secondhand smoke (SHS) exposure. No studies have assessed the impact of a smoking ban on SHS in enclosed areas outside separately ventilated, designated smoking rooms (DSRs). This study evaluated the overall impact of the Smoke-Free Ontario Act implemented May 31, 2006, on SHS in bars and coffee shops and the impact of banning DSRs on SHS outside DSRs. Air particulate matter (PM) and carcinogenic particulate polycyclic aromatic hydrocarbons (PPAH) were measured in May 2006 before the ban inside and outside DSRs in Toronto venues (13 coffee shops and 14 bars) that allowed smoking only in DSRs, and in Windsor venues (10 coffee shops and 10 bars) where smoking was allowed in shared spaces. Measurements were repeated 2 months post-ban. Air quality index values (AQIs) were calculated. Mixed model analysis was applied, taking into account measurement errors for repeated measures. Post ban, mean PM and PPAH levels were reduced by 87% (from 494 to 67 mm(2)/m(3)) and 94% (from 196 to 11 ng/m(3)), respectively, inside Toronto DSRs. Mean PM and PPAH levels were reduced by 10% (from 124 to 111 mm(2)/m(3)) and 46% (from 45 to 24 ng/m(3)), respectively, outside Toronto DSRs. In all Windsor venues, mean PM and PPAH levels were reduced by 83% (from 488 to 81 mm(2)/m(3)) and 90% (from 107 to 10 ng/m(3)), respectively. All reductions were statistically significant (p < 0.0001). In Toronto venues, the AQI was reduced from the "very unhealthy" range inside DSRs and the "moderate" range outside Toronto DSRs to the "good" range, and in Windsor venues from the "unhealthy for sensitive groups" range to the "good" range post-ban. Pre-ban PPAH levels including those outside Toronto DSRs may be associated with cardiovascular injury. DSRs did not provide adequate protection from SHS. The Smoke-Free Ontario Act produced a significant and firm reduction in SHS exposure in venues both with and without DSRs. PMID- 20017056 TI - Wearing the wrong size latex surgical gloves impairs manual dexterity. AB - Universal precautions mandate that health care workers wear gloves when dealing with patients, often in situations requiring a high level of technical skill. Although it seems obvious that wearing the wrong size gloves could impair or prolong tasks involving manual dexterity, the issue has not been formally studied. We tested the hypothesis that wearing the wrong size gloves impairs manual dexterity. We administered a grooved pegboard test to 20 healthy, paid, volunteer health care workers. The subjects performed the test with bare hands and while wearing their preferred size of latex surgical gloves, gloves that were a full size smaller, and gloves that were a full size larger. Each subject did three runs with each size glove and three runs with bare hands. The time necessary to insert pegs was measured with a stopwatch. Peg insertion time was not affected by wearing preferred size gloves (vs. bare-handed) but was increased 7-10% by gloves that were either too small or too large (both effects: P < 0.05 vs. preferred size; both P < 0.001 vs. bare-handed). The subjects reported that the too-small gloves limited hand motion or hurt their hands, whereas the too large gloves were clumsy but comfortable. Health care workers should wear gloves that fit properly when doing tasks that require manual dexterity. If the preferred size is unavailable, wearing gloves that are too large seems the best alternative. PMID- 20017057 TI - Development of a personal bioaerosol sampler based on a conical cyclone with recirculating liquid film. AB - This article describes the development of a novel, high-performance personal aerosol sampler intended to monitor occupational air pollution, specifically, microbial constituents. This prototype sampler has a horizontally positioned conical cyclone with recirculating liquid film and an ejection supply of adsorptive liquid into the inlet nozzle. Airborne pollutants were collected in the adsorptive liquid, thus improving the survivability of microbiological aerosol samples. Experimental modules of different dimensions were first evaluated. Based on the test results, a prototype sampler was fabricated and tested. Evaluation of the collection efficiency of the prototype unit indicated a higher than 90% collection efficiency for particles > 1.0 microm. The 50% cutoff diameter was between 0.70-0.75 microm. For assessment of the sampling process effect on the collected microorganisms, Bacillus thuringiensis was tested at a concentration of about 1.0 x 10(6) cells per cm(3). The viability in the prototype sampler decreased to 78% after 60 min of operation. PMID- 20017058 TI - Pupillometry reveals processing load during spoken language comprehension. AB - This study investigated processing effort by measuring peoples' pupil diameter as they listened to sentences containing a temporary syntactic ambiguity. In the first experiment, we manipulated prosody. The results showed that when prosodic structure conflicted with syntactic structure, pupil diameter reliably increased. In the second experiment, we manipulated both prosody and visual context. The results showed that when visual context was consistent with the correct interpretation, prosody had very little effect on processing effort. However, when visual context was inconsistent with the correct interpretation, prosody had a large effect on processing effort. The interaction between visual context and prosody shows that visual context has an effect on online processing and that it can modulate the influence of linguistic sources of information, such as prosody. Pupillometry is a sensitive measure of processing effort during spoken language comprehension. PMID- 20017059 TI - The five assessment issues you meet when you go to heaven. AB - In this article, based on an invited Master Lecture to the Society for Personality Assessment, I describe my wishes and predictions for the future of assessment. These include an outline and elaboration of the optimally written psychological report, use of actuarial procedures, treatment planning, model for a revised Rorschach, and use of higher technology procedures. I present a case to illustrate how the various issues can be translated into a client evaluation. PMID- 20017060 TI - Integrating methods to optimize circumplex description and comparison of groups. AB - Using the interpersonal circumplex as an exemplar, this article serves as a methodological primer for integrating techniques of group description and comparison when employing circumplex-based assessment instruments. Circular statistics (Mardia & Jupp, 1999) and the structural summary method (Gurtman & Balakrishnan, 1998) each offer unique and incrementally useful information when applied to group-level data on circumplex measures. Circular statistics offer a set of parameters that are conceptually similar to their linear equivalents (i.e., mean, variance, and confidence intervals). In interpersonal circumplex models, these parameters each provide specific information regarding substantive theme and group homogeneity and allow for the statistical comparison of groups based on the geometry of the circular model. In a similar fashion, the structural summary method for circumplex data provides a set of parameters that complement circular statistics by offering measures of the interpersonal prototypicality of the group profile, levels of profile differentiation and elevation, and a weighted measure of substantive theme. Used in conjunction, these methods offer more information than is available using either in isolation. We provide 4 examples to demonstrate the complementary information the 2 methods provide for assessments employing interpersonal circumplex measures. These examples will allow investigators to generalize the methods to other personality assessment domains in which circumplex models are utilized, such as emotion and vocational preference. [Supplementary materials are available for this article. Go to the publisher's online edition of the Journal of Personality Assessment for the following free supplemental resources: an Excel file that calculates the circular statistics and structural summary information described in this article using manually entered octant scores from up to 500 participants.] PMID- 20017061 TI - Anatomy of the White Bear Suppression Inventory (WBSI): a review of previous findings and a new approach. AB - The White Bear Suppression Inventory (WBSI; Wegner & Zanakos, 1994) was originally designed to assess people's inclination toward thought suppression. In this article, we provide a detailed review of previous findings on the structure of this instrument and present a study that took a new statistical approach. It involved an exploratory factor analysis of the French WBSI using the weighted least squares mean and variance estimator as well as parametric item response theory analyses. Results clearly supported a 2-factor structure with a "suppression" and an "intrusion" dimension. Follow-up regression analyses revealed that intrusion significantly predicted anxiety and depression scores, whereas suppression did not. PMID- 20017062 TI - An increase in early adolescent undercontrol is associated with the use of denial. AB - This is a longitudinal study of change in undercontrol and its relation to the use of defense mechanisms with participants from the Berkeley Guidance Study of the Institute of Human Development, University of California, Berkeley. I predicted that use of the immature defense of Denial, but not Projection or Identification, would be related at early adolescence to an increase in undercontrol as assessed from 2 independent measures. The assessment of Ego Undercontrol indicated that the majority of children decreased with age, but for those who increased at early adolescence, the increase was significantly related to the use of Denial. Similarly, assessment of Externalizing Behavior Problems at early adolescence indicated that an increase in Externalizing Problems was related to the use of Denial. In addition to indicating psychological immaturity, the use of Denial prevents these children from recognizing the negative impact of their undercontrolled behavior. PMID- 20017063 TI - The HEXACO-60: a short measure of the major dimensions of personality. AB - We describe the HEXACO-60, a short personality inventory that assesses the 6 dimensions of the HEXACO model of personality structure. We selected the 10 items of each of the 6 scales from the longer HEXACO Personality Inventory-Revised (Ashton & Lee, 2008; Lee & Ashton, 2004, 2006), with the aim of representing the broad range of content that defines each dimension. In self-report data from samples of college students and community adults, the scales showed reasonably high levels of internal consistency reliability and rather low interscale correlations. Correlations of the HEXACO-60 scales with measures of the Big Five factors were consistent with theoretical expectations, and convergent correlations between self-reports and observer reports on the HEXACO-60 scales were high, averaging above .50. We recommend the HEXACO-60 for use in personality assessment contexts in which administration time is limited. PMID- 20017064 TI - Impact of card rotation on the frequency of Rorschach reflection responses. AB - In this article, we describe the impact of Rorschach (Exner, 2003) card rotation and orientation preference on reflection responses. We anticipated exposure to sideways-orientated cards would facilitate landscape-type reflections, particularly for cards people find appealing to view sideways. When we examined 4 experimental conditions using an undergraduate sample, results in Experiments 1 (n = 123) and 2 (n = 38) showed that viewing the cards sideways produced a large increase in reflections. In Experiment 3 (n = 69), we examined preferences to view each card in a particular orientation. Cards producing higher rates of landscape reflections in the experimental conditions that encouraged turning were strongly correlated with preferences to view those cards sideways. The results imply reflections are in part a function of stimulus properties from viewing the card in a rotated orientation and not just the personal characteristics of the test taker. PMID- 20017065 TI - The interrater reliability of Elizur's hostility systems and Holt's aggression variables: a meta-analytical review. AB - We provide a meta-analytic review of interrater reliability for scoring the 2 most commonly studied Rorschach (2003) aggression measures: (a) The Elizur (1949) Hostility Scale and its main derivative scoring systems (Holtzman, Thorpe, Swartz, & Herron, 1961; Murstein, 1956) and (b) Holt's (1977, 2005) aggression variables. Substantial reliability was observed for both Elizur's hostility score (e.g., weighted mean summary score correlation = .91, N = 1,279) and Holt's aggression variables (e.g., weighted mean summary score correlation = .84, N = 226). These meta-analytic data suggest that like the contemporary variables included in Exner's Comprehensive System or Gacono and Meloy's (1994) extended aggression scores, the historically important Elizur scoring systems and Holt aggression variables can be scored reliably. [Supplementary materials are available for this article. Go to the publisher's online edition of the Journal of Personality Assessment for the following free supplemental resources: a document of results examining publication source, scoring system, amount of rater training, and severity of aggressive pathology in the sample as potential moderators of reliability.] PMID- 20017066 TI - Self-reported attachment patterns and Rorschach-related scores of ego boundary, defensive processes, and thinking disorders. AB - In this study, we addressed associations between self-reported attachment scales (anxiety and avoidance) and Rorschach (1921/1942) indexes indicating ego-boundary perception (barrier and penetration), use of projective identification, devaluation and splitting defenses, and Comprehensive System (Exner, 2005) scores that represent boundary blurring (incongruous and fabulized combinations). In this study, we extended the sample and findings described by Berant, Mikulincer, Shaver, and Segal (2005) using a nonclinical sample of 89 Israeli adults. We found significant associations between attachment orientations and Rorschach indexes dynamically related to anxiety. We also found a trend toward association between attachment orientation and avoidance. We discuss the theoretical and clinical implications of these findings. PMID- 20017067 TI - Are implicit motives "visible" to well-acquainted others? AB - In this study, we compared self-ratings and informant ratings of personality as predictors of implicit motives, need for achievement (nAch), and need for affiliation (nAff). A total of 120 participants wrote creative stories to 5 images from the Picture Story Exercise (Smith, 1992) and completed the Revised NEO Personality Inventory (Costa & McCrae, 1992). Three well-acquainted informants rated each participant's personality with the Big Five Inventory (John & Srivastava, 1999) and Saucier's (1994) Mini-Markers. Consistent with the study hypotheses, peer ratings of Conscientiousness significantly predicted nAch scores after controlling for word count and self-rated Conscientiousness. Contrary to hypotheses, peer ratings and self-ratings in all 5 domains did not significantly predict nAff scores. The findings are considered in the interest of bridging the gap between trait and motive concepts in personality assessment. PMID- 20017068 TI - Validating a new measure of self-complexity. AB - The construct of self-complexity has been recognized as important for understanding psychological adjustment. However, research in this area has been impeded by measurement problems. In particular, the most commonly used measure, Linville's (1985, 1987) H statistic, cannot assess this construct appropriately. In this article, we present a new measure designed to tap two components of self complexity separately: the number of self-aspects and the average distinction among them. We conducted a validation study with 131 Chinese college students. The results indicate that (a) this new measure is not confounded with conscious social desirability bias, (b) it has incremental validity beyond existing measures, and (c) the two components as assessed by this new measure are relatively independent and have different psychological functions. We discuss the implications of these findings for future studies of self-complexity and clinical practice. PMID- 20017069 TI - Assessing measurement and predictive invariance of the Toronto Alexithymia Scale 20 in U.S. Anglo and U.S. Hispanic student samples. AB - We collected data from a predominately Anglo American student sample in the Southeastern United States and a predominately Hispanic student sample in the Southwestern United States. Along with an assessment of internal consistency reliability, we examined measurement invariance of the Toronto Alexithymia Scale 20 (TAS-20) using confirmatory factor analysis. We also assessed the predictive invariance of the TAS-20. Results indicate that 2 of the 3 TAS-20 subscales demonstrated satisfactory internal consistency reliability across samples. Items from the TAS-20 subscales demonstrated measurement invariance of the latent means. The relationship between 2 measures of emotional dysfunction and the TAS 20 also demonstrated slope and intercept invariance, indicating equivalent validity. PMID- 20017071 TI - Association between paraoxonase activity and late saphenous vein graft occlusion in patients with coronary artery bypass grafting. AB - BACKGROUND: Coronary vein graft disease is an important contributor to the morbidity after coronary artery bypass grafting (CABG). Late occlusion of the graft is a serious complication that limits the use of the saphenous vein as a coronary bypass conduit. It is frequently encountered in old, degenerated vein grafts with advanced atherosclerotic plaque formation. Paraoxonase-1 (PON-1) is an HDL-bound enzyme which has anti-atherogenic properties and protects LDL cholesterol from oxidative modification. AIM: To examine the association between PON-1 activity and late saphenous vein graft occlusion. METHODS: Thirty-eight patients who had at least one occluded saphenous vein graft (group 1; 12 females, 26 males) and 41 patients who had a patent saphenous vein graft (group 2; 7 females, 34 males) were enrolled in this study. Paraoxonase activity was measured spectrophotometrically. RESULTS: The mean PON-1 activity in group 1 was significantly lower than in group 2 (74.1 +/- 52.1 vs. 114.4 +/- 90.9 U/l, p = 0.02). The mean platelet volume was significantly higher in group 1 than group 2 (8.8 +/- 1.6 vs. 8.2 +/- 1.1 fl, p = 0.04). Multiple logistic regression analysis showed that only PON-1 activity (beta = 0.011, p = 0.042) was an independent predictor of late occlusion of a saphenous vein graft. CONCLUSIONS: Our results show that PON-1 activity is lower in patients with late saphenous vein graft occlusion. Reduced PON-1 activity may lead to acceleration of saphenous vein graft occlusion. PMID- 20017072 TI - M-mode speckle tracking--a novel echocardiographic approach to assess left ventricular torsional deformation. AB - BACKGROUND: Speckle tracking echocardiography (STE) requires special image processing for complex assessment of left ventricular (LV) function, including strain and rotation. AIM: To evaluate a novel M-mode STE technique as a readily applicable approach, providing potential insights into LV deformation. METHODS: Fifty one patients (mean age 52 +/- 14 years, 24 women) with normal or impaired LV function (mean LVEF 51 +/- 16%, range between 14% and 66%) were studied. Left ventricular rotation at mitral valve and apical level was measured using dedicated conventional STE software. Grey-scale short axis digital loops at mitral valve level were also used to obtain M-mode STE images, presenting the movement of LV wall speckles along the cursor, with the LV cavity left out. Then, the distance of peak systolic shift (PSS) was measured for one selected speckle. RESULTS: Mean rotation at mitral valve level was 7.4 +/- 3.1 degrees , apical rotation was 7.6 +/- 6.4 degrees and LV torsion was 14.9 +/- 7.1 degrees . Measurement of PSS with a novel M-mode STE approach was feasible in all patients, producing a mean value of 7 +/- 2 mm. Subsequently, two subsets with PSS of less than 7 mm and PSS equal to or above 7 mm showed a significant difference between mean LV torsion of 10.7 degrees and 17.0 degrees , respectively (p = 0.002). In the ROC analysis, PSS cut-off value of less than or equal to 7 mm yielded a sensitivity of 92% and a specificity of 61% to predict LV torsion < 10.7 degrees (p < 0.0001). CONCLUSIONS: M-mode STE appears to be a feasible approach to detect movement of speckles and to measure PSS within the LV inferior and septal wall in short axis view images. PSS cut-off value of less than 7 mm indicates significantly lower LV torsion. Further studies are required to assess the potential role of M-mode STE in cardiac imaging. PMID- 20017073 TI - Quality of life in young, professionally active men undergoing on-pump coronary artery bypass grafting--short-term follow-up results. AB - BACKGROUND: The impact of coronary artery bypass grafting (CABG) on the quality of life (QoL) is one of the important measures of the efficacy of the procedure. This issue in young, professionally active male patients has not been extensively studied. AIM: To assess QoL before and after on-pump CABG, and before and after cardiac rehabilitation in young men with low operative risk. METHODS: The study group comprised 50 men aged 54.4 +/- 5.6 years who were professionally active before the surgery. The QoL was assessed on the basis of the MacNew questionnaire (in points). We analysed QoL changes and effects of basic demographic and peri operative data on QoL during a short-term follow-up. RESULTS: All components of QoL deteriorated shortly after CABG: emotional - from 4.97 +/- 0.96 to 4.66 +/- 1.0 (p = 0.03); physical - from 4.49 +/- 1.1 to 4.2 +/- 1.2 (p = 0.02); and social - from 4.68 +/- 1.0 to 4.47 +/- 1.1 (p = 0.1). Pre-operative physical and social QoL positively correlated with age (r = 0.45 and r = 0.37, respectively) and left ventricular ejection fraction (LVEF) (r = 0.49 and r = 0.48, respectively). However, there was a negative impact of history of myocardial infarction on physical QoL (p < 0.05). A negative influence of cardiopulmonary bypass time (r = -0.45) and cross-aortic clamp time (r = -0.36) on physical QoL was also noted. The QoL values were also influenced by class of angina symptoms (R = -0.33 / -0.42), total drainage (r = -0.11 / -0.34) and quantity of grafts (R = -0.35 / -0.42). During rehabilitation, QoL significantly improved: emotional - from 5.29 +/- 0.92 to 5.96 +/- 0.9 (p = 0.01); physical - from 4.66 +/- 1.1 to 5.42 +/- 1.2 (p < 0.01); and social - from 4.69 +/- 1.2 to 5.65 +/- 1.1 (p < 0.01). The QoL during rehabilitation was correlated with baseline peri-operative risk (for logistic EuroSCORE algorithm r = -0.21 / -0.31 and for EuroSCORE R = 0.47 / -0.89). Significant determinants of some components of QoL were also LVEF (r= 0.26 / 0.47), morphological blood parameters (r = 0.37 / 0.43), baseline CCS class (R = 0.31 / 0.58), age (r = -0.41 / -0.83), and extent of surgery defined by cardiopulmonary bypass time, cross-aortic clamp duration and total drainage. CONCLUSIONS: Quality of life in young, professionally active men significantly deteriorates a few days after on-pump CABG but systematically improves during the next weeks, particularly after rehabilitation. Pre-operative QoL correlates positively with age and LVEF, and negatively with a history of myocardial infarction. Age, pre-operative risk, angina symptoms and the extent of surgery have negative effects on physical QoL after CABG during short-term observation. PMID- 20017074 TI - Investigation of SERPINE1 genetic polymorphism in Macedonian patients with occlusive artery disease and deep vein thrombosis. AB - BACKGROUND: Raised SERPINE1 plasma levels are related to a 1-bp guanine deletion/insertion (4G5G) polymorphism in the promoter of the SERPINE1 (plasminogen activator inhibitor 1 - PAI1) gene. Evidence suggested that the plasma levels of SERPINE1 modulate the risk of coronary artery disease; furthermore, that the 4G5G polymorphism affects the expression of the SERPINE1 gene. AIM: To analyse association of SERPINE1 polymorphism with occlusive artery disease (OAD) and deep venous thrombosis (DVT) in Macedonians in order to investigate its role as a part of candidate genes in different vascular diseases in Macedonians. METHODS: Investigated groups consisted of 82 healthy patients, 75 with OAD, and 66 with DVT. Blood samples were collected after written informed consent was obtained, and DNA was isolated from peripheral blood leukocytes. Identification of SERPINE1 polymorphism was done with CVD StripAssay (ViennaLab, Labordiagnostica GmbH, Austria). The population genetics analysis package, PyPop, was used for analysis of the SERPINE1 data. Pearson's P-values, crude odds ratio and Wald's 95% CI were calculated with Bonferroni corrected p value. RESULTS: The frequency of 4G allele for SERPINE1 was 0.538 for DVT, 0.555 for healthy participants, and 0.607 for OAD. The frequency of 5G allele for SERPINE1 was the smallest in patients with OAD (0.393) and was higher in healthy participants (0.445), and patients with DVT (0.462). Test of neutrality (Fnd) showed negative value, but was significantly different from 0 for SERPINE1 in healthy participants (p of F = 0.041) and in patients with DVT (p of F = 0.030). SERPINE1 genotypes in healthy participants and patients with OAD were not in Hardy Weinberg proportions (p = 0.019 and 0.001, respectively). No association between SERPINE1 polymorphisms and OAD or DVT was found. CONCLUSION: There is no significant relationship between SERPINE1 polymorphisms and occlusive artery disease or deep venous thrombosis in Macedonian population. PMID- 20017075 TI - Analysis of 12-lead electrocardiogram in top competitive professional athletes in the light of recent guidelines. AB - BACKGROUND: One of the most important aims of modern sports cardiology is prevention of sudden cardiac death among athletes. Adequate pre-participation screening is a crucial part of prevention, however, current ACC, AHA or ESC guidelines are not uniform in this context. There is recently ongoing discussion on implementation of 12-lead ECG to the screening protocol. AIM: To assess the prevalence of alterations of resting 12-lead ECG in a population of top-level professional athletes - members of the Polish Olympic Team - using recently accepted criteria. METHODS: During the period of intensive training before the Summer Olympic Games in Beijing (2008), a 12-lead, resting ECG was performed in 73 members (20 women and 53 men) of the Polish Olympic Team. Commonly accepted criteria were used to assess the ECG, and alterations were divided into two groups according to recent publications: group I - 'benign', common - thought to be consistent with the athlete's heart syndrome (i.e.: sinus bradycardia, 1st degree atrioventricular block, early repolarisation, right bundle branch hemiblock, isolated signs of left ventricular hypertrophy); and group II - 'suspected', uncommon - which may occur due to organic heart disease (i.e. complete bundle branch block, ventricular arrhythmia, inverse T wave or pathological QRS axis deviation). RESULTS: Completely normal ECG was present in 11% of those examined, common (group I) findings were observed in 65% and 'suspected' (group II) in 23%. The most commonly occurring 'benign' findings were bradycardia incomplete, right bundle branch block and isolated left ventricular hypertrophy, found in 75, 71 and 41%, respectively. From 'suspected' (group II) the most frequent was left posterior fascicular hemiblock, present in 10% of those examined; other findings were complete right bundle branch block, left atrial hypertrophy, inverse T waves and left anterior fascicular hemiblock in single cases. CONCLUSIONS: 1. Most of the observed alterations in resting ECG of professional athletes belong to the 'common' group and result from adaptation to exercise. 2. Frequent occurrence of left posterior fascicular hemiblock, which is thought to be 'potentially malignant', requires further investigation. PMID- 20017076 TI - The twiddler's plus syndrome--a case report. AB - We present a case of a 43-year-old male with an extreme form of the twiddler's syndrome - almost complete lead dislodgement associated with their fracture. PMID- 20017077 TI - [Myocardial infarction after carbon monoxide poisoning]. AB - Carbon monoxide (CO) exposure is a common cause of unintentional poisoning. Clinical manifestation usually involve central nervous and cardio-vascular systems as brain and heart are very sensitive to hypoxia. Myocardial infarction pathogenesis during CO intoxication is complex. Generalised tissue hypoxia and increased thrombotic tendency may play an important role. A case of ST-segment elevation myocardial infarction following CO poisoning in a 58-year-old male is presented. PMID- 20017078 TI - [Severe mitral regurgitation following blunt trauma of the chest as a cause of over heart failure]. AB - A case of a 33-year-old previously healthy man who due to a car accident suffered from severe multiorgan injuries, is presented. Two months following the injury, the patient developed symptoms of severe heart failure. A significant mitral regurgitation was confirmed together with a torn chordate tendineae being the main cause of the valve dysfunction. Mitral valve plasty was performed using a Carpentier Edwards mitral ring. After 14 months, the patient was selected for reoperation due to the endocarditis process which also affected the mitral ring. The patient had a St. Jude Medical 27-mm mitral valve implanted. PMID- 20017079 TI - [ST-segment elevation myocardial infarction in a 78-year-old patient with post traumatic central nervous system haemorrhage]. AB - A case of a 78-year-old patient with ST-segment elevation myocardial infarction and concomitant post-traumatic central nervous system hemorrhage is described. The patient underwent successful primary percutaneous coronary intervention with aspiration catheter without antiplatelet and antithrombothic treatment. Dissimilarities in acute coronary syndromes treatment in patients with major bleeding (central nervous system hemorrhage) are discussed. PMID- 20017080 TI - [Proinflammatory capacity of adipose tissue--a new insights in the pathophysiology of atherosclerosis]. PMID- 20017081 TI - [The importance of periodontal treatment in patients with cardiovascular diseases]. PMID- 20017082 TI - [Recommendations for the standardization and interpretation of the electrocardiogram according to the American Heart Association Electrocardiography and Arrhythmias Committee and the Heart Rhythm Society 2007 and 2009 - new standards]. PMID- 20017083 TI - [Unusual left-to-right shunt]. AB - A case of a 58-year-old woman with restrictive cardiomyopathy and unusual interatrial left-to-right shunt is presented. Transthoracic echocardiography suggested the coronary artery fistula draining into right atrium. Transoesophageal echocardiography revealed patent foramen ovale with concomitant left-to-right shunt. Increased pressure within the both enlarged atria caused the inefficiency of the valve of PFO and interatrial leak leading to 'acquired form' of an atrial septal defect. PMID- 20017084 TI - [Optical coherence tomography-guided stent implantation]. AB - A case of 51-year-old woman who was admitted to Department of Cardiology and Internal Medicine in Bydgoszcz due to unstable angina pectoris with elevated level of troponin I is presented. The patient underwent optical coherence tomography-guided angioplasty of the proximal left anterior descending artery with an implantation of everolimus-eluting stent. Optical coherence tomography (OCT) visualized a plaque or thrombus prolaps between stent struts that was undetectable by intravascular ultrasound. A postdylatation with a bigger balloon was performed. Final OCT imaging revealed a well apposed and expanded coronary stent without any tissue prolaps between struts. PMID- 20017085 TI - [Sixty four-slice computed tomography unmasked unexpected source of atrial arrythmias--a case report]. AB - We describe a 23-year-old man with drug refractory, left atrial arrhythmias (paroxysmal atrial tachycardia, runs, pairs and single extra beats) in the area of remnant of left upper pulmonary vein ostium (in fact, this vein). The superior drained into vena innominata/vena cava superior. The patient underwent three unsuccessful catheter ablations in another center. Computed tomography scan of left atrium and pulmonary veins revealed this rare congenital anomaly. This information was crucial to perform successful ablation in the area of remnant and distinguish left atrial appendage from the remnant area. During 6 month follow-up the patient remained free of arrhythmia. PMID- 20017086 TI - [POL-PAVTI--Polish report on transcatheter pulmonary artery valve implantation of Melody-Medtronic prosthesis in the first 14 patients in Poland]. AB - AIM: To assess the early results of the pulmonary artery valve transcatheter implantation (PAVTI) in pts included into POL-PAVTI registry. Detailed medical and economic analyses were performed. METHODS: Pulmonary artery valve implantation was performed in 14 pts (9 men), aged 16-31 (mean 24.6 +/- 4.8) years, with pulmonary homograft dysfunction after total repair of tetralogy of Fallot (4 pts), pulmonary atresia (2 pts), pulmonary stenosis (1 pt), common arterial trunk type I (1 pt), Ross procedure (3 pts) and TGA - Rastelli operation (3 pts). Eleven pts underwent in the past 2-5 surgical or/and catheter interventions. Indication for PAVTI was based on clinical evaluation and echocardiographic studies. Assessment of morphological and functional features of the right ventricle (RV) and homograft with the use of cardiac magnetic resonance (CMR) was performed in 10 cases. Pulmonary stenosis (max. pulmonary gradient 32 119, mean 72 +/- 28 mmHg) was observed in 13 pts and/or significant pulmonary regurgitation in 10 pts. The procedure was performed in general anesthesia. The deployment of a valved stent in the pulmonary valve position was preceded by a metal stent implantation. Results were evaluated by echocardiography two days after the procedure and one month later. Four patients were evaluated 6 months after procedure. RESULTS: Time of the procedure varied 60-190 (mean 127 +/- 35) min, time of fluoroscopy ranged 12-31 (mean 21 +/- 11) min. PAVTI was successfully performed in all pts without serious complications. Patients were discharged from the hospital 48-293 (mean 120 +/- 71) h after procedure. Significant reduction of pulmonary gradient after the procedure assessed by echocardiography was observed on the second day (20-60, mean 38 +/- 12 mmHg, p < 0.0001) and one month (19-52, mean 34 +/- 9 mmHg, p < 0.0001). Mild pulmonary regurgitation was observed in 2 pts. In 5 pts evaluated 6 months after procedure haemodynamic parameters were unchanged; no late complications were observed. Average cost of the procedure including a price of the valve (82 000 PLN) was 98 000 PLN. CONCLUSIONS: Pulmonary artery valve transvascular implantation is an effective and safe method of non-surgical treatment for patients with homograft dysfunction. Cost-effectiveness is approvable. PMID- 20017087 TI - [Percutaneous transfemoral aortic CoreValve(R) implantations in high risk patients--another Polish good experience]. PMID- 20017088 TI - Robotic versus manual control in magnetic steering of an endoscopic capsule. AB - BACKGROUND AND STUDY AIMS: Capsular endoscopy holds promise for the improved inspection of the gastrointestinal tract. However, this technique is limited by a lack of controlled capsule locomotion. Magnetic steering has been proposed by the main worldwide suppliers of commercial capsular endoscopes and by several research groups. The present study evaluates and discusses how robotics may improve diagnostic outcomes compared with manual magnetic steering of an endoscopic capsule. MATERIALS AND METHODS: An endoscopic capsule prototype incorporating permanent magnets was deployed in an ex vivo colon segment. An operator controlled the external driving magnet manually or with robotic assistance. The capsule was maneuvered through the colon, visualizing and contacting targets installed on the colon wall. Procedure completion time and number of targets reached were collected for each trial to quantitatively compare manual versus robotic magnetic steering ( T-test analysis with P = 0.01). Then, through a set of in vivo animal trials, the efficacy of both approaches was qualitatively assessed. RESULTS: In ex vivo conditions, robotic-assisted control was superior to manual control in terms of targets reached (87 % +/- 13 % vs 37 % +/- 14 %). Manual steering demonstrated faster trial completion time (201 +/- 24 seconds vs 423 +/- 48 seconds). Under in vivo conditions, the robotic approach confirmed higher precision of movement and better reliability compared with manual control. CONCLUSIONS: Robotic control for magnetic steering of a capsular endoscope was demonstrated to be more precise and reliable than manual operation. Validation of the proposed robotic system paves the way for automation of capsular endoscopy and advanced endoscopic techniques. PMID- 20017089 TI - [Burn treatment in the renaissance by Fabricius Hildanus--a historical appraisal]. AB - A first comprehensive textbook on burns treatment was written in 1607 by Fabry of Hilden (Fabricius Hildanus). This monograph describes cause, diagnosis, treatment, and complications of burn injuries. Besides a variety of topical ointments with promising herbal ingredients like onion and camphor, surgical procedures like necrectomies, escharotomies and syndactely treatment are described for the first time ever. Scar management including splinting devices is another interesting topic. Some of the therapeutic procedures are still valid today. Thus this renaissance piece of medical writing belongs to the pedigree of surgery. PMID- 20017090 TI - Role of short chain fatty acids in mesenteric ischemia reperfusion injury in rats. AB - AIM: The aim of this study was to evaluate the role of short chain fatty acid administration in mesenteric ischemia reperfusion in rats. METHODS: Sixteen 60 day-old male Wistar rats were divided into two groups of eight each. In group I (n=8) normal saline (10 ml/kg) was injected into the ileal lumen. Ischemia reperfusion was achieved by clamping the superior mesenteric artery (SMA) for 30 min and then releasing it for another 30 min. The ileum was harvested for histopathological examination. In group II (n=8) a mixture of short chain fatty acids (10 ml/kg) was injected into the ileal lumen. Ischemia reperfusion was achieved in a similar fashion and the ileum harvested for histopathological examination. Injury was graded according to Chiu's score. RESULTS: The ileum in rats from group II showed a lower injury score (0.013+/-0.354) compared to ileum from rats in group I (4.631+/-0.521), and this difference was significant (p<0.001). CONCLUSION: Short chain fatty acids significantly decreased the degree of reperfusion injury in the gut in a rat model of mesenteric ischemia reperfusion. PMID- 20017091 TI - Teratoid Wilms' tumor with hypertension treated with partial nephrectomy: case report with literature review. PMID- 20017092 TI - Robot-assisted laparoscopic hemihysterectomy for a rare genitourinary malformation with associated duplication of the inferior vena cava--a case report. PMID- 20017093 TI - ["We owe our patients the endoprosthesis registry" (interview by Dr. Bernhard Epping)]. PMID- 20017095 TI - [Conservative symptomatic treatment--NSAIDs: protection or damage to the cartilage?]. PMID- 20017096 TI - [Conservative treatment of femoral head necrosis--alendronate as a treatment for avascular femoral head necrosis]. PMID- 20017097 TI - [Subtalar arthrodesis--heel bone fractures--always operate first?]. PMID- 20017098 TI - [Shaft fractures of the upper and lower leg--operating day and night?]. PMID- 20017099 TI - Endoscopic anatomy for transnasal transsphenoidal pituitary surgery in the presence of a persistent trigeminal artery. AB - OBJECTIVE: Endoscopic procedures are becoming increasingly important for transnasal transsphenoidal approaches to the skull base and particularly for pituitary surgery. A persistent trigeminal artery (PTA) is rare. Its presence, if it goes unnoticed or if the surgeon is not aware of such a variant, may endanger the success of surgery. METHOD: During an endoscopic inspection using a supraorbital approach in a fresh cadaveric specimen in which the arteries had been injected with latex glue, the presence of an anomalous intracranial artery, suggestive for PTA, was disclosed. The specimen was then fixed and a CT scan with 3D reconstruction of the circle of Willis was done to evaluate the imaging of such an anatomical variation. Thereafter an endoscopic transsphenoidal approach to the pituitary fossa was performed, to verify the endoscopic anatomy. RESULTS: The performed CT scan allowed visualization of the entire course of the anomalous vessel, confirming a PTA. During the endoscopic transsphenoidal approach, the presence of the vascular anomaly, altering the bony bulging of the internal carotid artery on the lateral side of the sphenoidal roof, was disclosed. The parasellar course of the PTA could be exposed by drilling the overhanging bone. The presence of the anatomical variant did not interfere with surgical manoeuvres and the procedure, simulating a transsphenoidal approach to the pituitary, could be safely completed. CONCLUSION: Variants such as PTA are rare and routine preoperative imaging for pituitary procedures does not always include studies to detect this vascular anomaly. The occasional intraoperative detection of a PTA during an endoscopic transsphenoidal procedure can be managed; almost any surgical manipulation is possible and pituitary surgery can be successfully completed, provided the surgeon is aware of the possible existence of this variant and its irregular anatomical course. However, the presence of a PTA may have dramatic consequences if surgery is directed to the lateral parasellar region, as for intracavernous lesions; in these cases a complete neuroradiological study including MRI-angiography and possibly CT-angiography is advised. PMID- 20017100 TI - Nandrolone decanoate increases satellite cell numbers in the chicken pectoralis muscle. AB - The anabolic androgenic steroid nandrolone decanoate has minimal androgenic effects and, thus, is widely used to induce muscle hypertrophy in both patients and athletes. Although increases in satellite cell numbers and satellite cells giving rise to new myonuclei are associated with hypertrophy in many experimental models, the relationship between nandrolone and satellite cells is poorly understood. Here we test the hypothesis that nandrolone administration is associated with an increase in satellite cell numbers in muscle. Nandrolone was injected at weekly intervals for four weeks into the right pectoralis muscle of female white leghorn chickens aged 63 days post hatch. Age/size/sex matched control birds received saline injections. The contralateral pectoralis was excised for study from each control and nandrolone treated bird. An antibody against Pax7 and immunocytochemical techniques were used to identify satellite cells. Nandrolone significantly increased mean pectoralis mass by approximately 22%, and mean fiber diameter by about 24%. All satellite cell indices that were quantified increased significantly in chicken pectoralis with administration of nandrolone. Nandrolone injected birds had on average higher satellite cell frequencies (#SC nuclei/all nuclei within basal lamina), number of satellite cells per millimeter of fiber, and satellite cell concentrations (closer together). Myonuclei were further apart (less concentrated) in nandrolone injected muscle. However, an overall increase in myonuclear numbers was revealed by a significantly greater mean number of myonuclei per millimeter of fiber in nandrolone injected muscle. Our results suggest that satellite cells may be key cellular vectors for nandrolone induced muscle fiber hypertrophy. PMID- 20017101 TI - The effects of eicosapentaenoic acid on the endothelium of the carotid artery of rabbits on a high-cholesterol diet. AB - The preventive and therapeutic effects of eicosapentaenoic acid (EPA) on diet induced hyperlipidemia in rabbits have been investigated. Eighteen New Zealand rabbits were randomly divided into three groups of 6 subjects each; experimental group-I (EG-I) was administered a cholesterol rich diet, experimental group-II (EG-II) was treated with EPA (300 mg/kg/d) following a cholesterol-rich diet and the control group (CG) had a standard diet. Blood samples were collected at day 0 and at the 4th and 12th weeks of EG-II to obtain serum levels of total cholesterol (TC), high density lipid-cholesterol (HDL-C), low density lipid cholesterol (LDL-C) and triglyceride (TG). From each group tissue samples were collected from the carotid artery for immunohistochemistry and electron microscopy. Our results showed that EPA could significantly lower (p<0.001) serum TC, LDL-C, HDL-C and TG levels with a reduction of 35%; 55%; 44% and 51%, respectively. Scanning and transmission electron microscopy results revealed that endothelial damage was more prominent in EG-I when compared to EG-II. The ruptured endothelial lining and damaged cellular surface was increased in EG-I when compared to EG-II. Ultrastructural observations showed that after EPA treatment, the degeneration and cellular surface damage on the endothelium were also decreased. These biochemical and ultrastructural results suggest that EPA is a potential drug which significantly lowers the serum lipid profile and partially repairs endothelial dysfunction due to hyperlipidemia. PMID- 20017102 TI - Morphometric study of healthy jejunal and ileal mucosa in adult and aged subjects. AB - Small intestine mucosa is often affected with malabsorptive, autoimmune and inflammatory pathological processes. However, morphometric data on the healthy human small intestine mucosa, especially ileum, are scarce. We aimed to obtain histoquantitative data on the healthy jejunal and ileal mucosa and assess the effects of gender and ageing on these parameters. Computer-aided morphometric analysis was performed on 24 jejunal and 25 ileal biopsy samples collected upon routine endoscopy screening of healthy persons with a family history of intestinal malignancy. Subjects were distributed in four groups according to age and sex: adult (<60 years) and elderly (>60 years) males, and adult (<60 years) and elderly (>60 years) females. Results were statistically analyzed with Mann Whitney U test. Jejunal mucosal thickness was significantly reduced in elderly subjects (p<0.05), especially in elderly females compared to adult ones (p<0.05). Jejunal villi were significantly wider in adult than in the elderly subjects (p<0.05), whereas ileal villi were significantly wider in elderly compared to adult subjects (p<0.01) and in male compared to female subjects (p<0.05). No statistically significant differences were found in other histoquantitative parameters (mucosa epithelium height, crypt numerical density, villous height, crypts and villous perimeter, diameter and epithelium height) of jejunal and ileal mucosa. This study provides complete morphometric data on the healthy human jejunum and the first relevant data on the healthy ileal mucosa, thus representing a valuable morphometric reference for future histoquantitative studies of human small bowel mucosa in both healthy and disease affected individuals. PMID- 20017103 TI - The distribution of vasotocin and mesotocin immunoreactivity in the hypothalamic magnocellular neurosecretory nuclei of the Saharan herbivorous lizard, Uromastix acanthinurus Bell, 1825 (Sauria-Agamidae). AB - An immunohistochemical study of the magnocellular neurosecretory nuclei was performed in the hypothalamus of the desert lizard Uromastix acanthinurus using polyclonal antibodies against arginine vasotocin (AVT), mesotocin (MST) and neurophysins I and II (NpI, NpII). AVT- and MST-immunoreactivities were localized in individual neurons of the supraoptic, periventricular, and paraventricular nuclei and in scattered neurosecretory cells. The supraoptic nuclei (SONs) can be subdivided into rostral, medial and caudal portions. The rostral portion of the SONs was called the SON-ventral aggregation (V SON) because the neurosecretory neurons are present in the ventral part of the hypothalamus along the optic chiasma (OC). Their perikarya and fibres were only AVT-ir. The medial part of the SONs was constituted of two clusters of neurosecretory neurons located in the two lateral ends of the OC to form the SON-lateral aggregations (L SON). In the caudal end of the last one, some MST-ir perikarya appeared. The caudal part of the SONs was constituted of a dorso-lateral aggregation (D SON) of ir-neurons spreading over the lateral forebrain bundle (LFB). AVT- and MST- perikarya were observed in this caudal portion of the SONs, AVT-ir neurons being more numerous. AVTergic and MSTergic magnocellular neurons were present in the periventricular nuclei (PeVNs). Parvocellular and magnocellular AVT- and MST-ir were observed in the paraventricular nuclei (PVNs). The fibres emerging from the magnocellular neurons which belong to these nuclei and the scattered cells ran along the hypothalamic floor and entered the median eminence (ME) to end in the neural lobe of hypophysis. As a rule, immunoreactivity was also observed in all the regions of the forebrain with vasotocinergic and mesotocinergic perikarya and fibres. The immunoreactive distribution was similar to that described in other reptiles. PMID- 20017104 TI - Expression and distribution of the intermediate filament protein nestin and other stem cell related molecules in the human olfactory epithelium. AB - The olfactory epithelium (OE) is unique in regenerating throughout life and thus is an attractive target for examining neurogenesis. The nestin protein was shown to be expressed in the OE of rodents and is suggested to be essentially involved in the process of regeneration. Here we report the expression and distribution of nestin in the human OE at RNA and protein level. Moreover, we analysed the expression profiles in dependence on age and olfactory capacity. After sinus surgery, biopsies were taken from the olfactory epithelium of 16 patients aged 20 80 years with documented differences in their olfactory function. Our studies revealed that nestin is constantly detectable in the apical protuberances of sustentacular cells within the human OE of healthy adults. Its expression is not dependent on age, but rather appears to be related to the olfactory function, as a comparison with specimens obtained from patients suffering either from persistent anosmia or hyposmia suggests. Particularly, in the course of dystrophy, often accompanied with impaired olfaction, nestin expression was occasionally decreased. Contrarily, the expression of the p75-NGFR protein, a marker for human OE basal cells, was not altered, indicating that at least in the tested samples olfactory impairment is not connected with abnormalities at the basal cell level. These observations emphasize an essential role of nestin for the process of regeneration, and also highlight this factor as a candidate marker for sustentacular cells in the human olfactory epithelium. PMID- 20017105 TI - Tubular epithelial cell and podocyte apoptosis with de novo sirolimus based immunosuppression in renal allograft recipients with DGF. AB - Sirolimus is associated with prolonged delayed graft function (DGF) following renal transplantation and exacerbation of proteinuria. We assessed renal allograft biopsies from DGF patients treated with de novo sirolimus (n = 10) for renal tubular cell and podocyte apoptosis and expression of activated caspase-3, Bcl-2, and mTOR and compared them to biopsies from DGF patients not receiving sirolimus (n = 15). Both groups received mycophenolate mofetil, prednisone and antibody induction. Apoptosis was assessed using terminal deoxynucleodidyl transferase mediated dUTP nick end labeling (TUNEL) staining. Caspase-3, Bcl-2, and mTOR expression were assessed by immunohistochemistry. Sirolimus treated patients had 334+/-69 TUNEL positive cells per 5 high power fields compared to 5.5+/-2.9 TUNEL positive cells in control patients (p<0.001). The number of TUNEL positive cells correlated with tubular architectural disruption. Expression of activated caspase-3, Bcl-2, or activated mTOR did not differ between groups. 60% of biopsies from sirolimus treated patients compared to 7% of biopsies from controls showed diffuse podocyte apoptosis (p = 0.007). There was no podocyte expression of activated mTOR, activated caspase-3, or Bcl-2 in either group. These data suggest that DGF patients treated with sirolimus have increased renal tubular cell apoptosis and podocyte apoptosis. PMID- 20017106 TI - In vitro antigen-specific cytotoxic T cell response against esophageal carcinoma cells induced by HPV18E7-transfected dendritic cells. AB - Human Papillomavirus (HPV)-associated esophageal carcinoma (EC) is a high incidence tumor worldwide. Dendritic cell (DC)-based tumor vaccine is considered an alternative therapy to treat EC. Here we developed a DC-based vaccine by transfecting cord blood CD34+ stem cell-derived DC with HPV18E7 gene, observed its biological characteristics and the antigen-specific T-cell cytotoxicity on EC cells induced by HPV18E7-DC in vitro. Our results showed that 1) HPV18E7 gene transfer did not change the typical morphology of mature DC, 2) the representative phenotypes of mature DC (CD80, CD86, and CD83) were highly expressed in HPV18E7- DC (81.6%, 80.5%, and 86.6%, respectively), 3) the expression level of 18E7 protein in HPV18E7-DC was 47.5%, and 4) the specific cytotoxicity against EC cells was significantly higher than that in controls (p<0.01). This study indicates the possibility of a DC-based immunotherapy in HPV associated EC. PMID- 20017107 TI - Lectin-binding pattern of Senegalese sole Solea senegalensis (Kaup) testis. AB - The localization and characterization of oligosaccharide sequences in the testis of Senegalese sole Solea senegalensis was investigated using 12 lectins in combination with KOH saponification and sialidase digestion (K-s). The interstitial compartment contained all the sugar residues investigated, those bearing oligosaccharides terminating with sialic acid (Neu5Ac) alpha2,3Galbeta1,4GlcNAc, Neu5AcGalNAcalpha1,3(LFucbeta1,2) Galbeta1,3/4GlcNAcbeta1 and GalNAcalpha1,3(LFuc1,2) Galbeta1,3/4GlcNAcbeta1 being more abundant in the medullar region than in the cortex. The melano-macrophage centres found in the interstitial compartment displayed glycans terminating with Galbeta1,3GalNAc. The basal lamina separating the germinal and interstitial compartments exhibited glycans with terminal/internal mannose, internal betaGlcNAc, and terminal Neu5Acalpha2,6Gal/GalNAc, and Neu5AcGalbeta1,3GalNAc, Galbeta1,3GalNAc (PNA), Galbeta1,4GlcNAc, GalNAc, alphaGal, and alphaL-Fuc. In the germinal compartment, the Sertoli cells expressed only glycans terminating with Neu5Acalpha2,3Galbeta1,4GlcNAc in the apical and supra-nuclear lateral surface of the spermatonial cysts located in the distal part of the seminiferous lobules. Primary spermatocytes exhibited oligosaccharides terminating with Galbeta1,3GalNAc and alphaGalNAc in the cytoplasm and nucleus, respectively. The spermatids contained highly mannosylated glycans terminating with GalNac, alphaGal, and alphaL-Fuc. The head of spermatozoa expressed a more complex glycosylation pattern characterized by the additional presence of oligosaccharides terminating with Neu5Acalpha2,3Galbeta1,4GlcNAc, Neu5AcGalbeta1,3GalNAc, Neu5AcGalNAcalpha1,3(LFuca1,2)Galbeta1,3/4GlcNAcbeta1, GalNAcalpha1,3(LFucalpha1,2)Galbeta1,3/4GlcNAcbeta1. The comparison with previous lectin histochemical studies carried out in other fish species reveals a specific glycosylation pattern of Senegalese sole testicular structures and spermatozoa head. PMID- 20017108 TI - Small cell carcinoma of the urinary bladder. AB - Small cell carcinoma of the urinary bladder (SCCUB) is a rare and aggressive cancer of the bladder. SCCUB is part of neuroendocrine family of tumors that affect several organ systems including respiratory, gastrointestinal and male and female genitourinary tract. SCCUB affect males predominantly with common risk factors include smoking, bladder calculi, bladder manipulation, and chronic cystitis. Prognosis of SCCUB remains poor due to high metastatic potential and lack of symptoms in earlier stages of the disease. Pathogenesis of the disease is linked to loss of genetic material, hypermethylation of tumor suppressors and at times amplification of the chromosomal regions carrying oncogenes. Majority of cases are treated with local resection of the tumor with neoadjuvant or adjuvant platinum-based chemotherapy regimen. Radiation therapy is used as alternative to radical cystectomy or as palliative measure. This article provides epidemiology, molecular pathogenesis, histochemistry, and current management options for SCCUB. Furthermore we reviewed all recent studies involving advancement in targeted molecular therapy for neuroendocrine tumors. PMID- 20017109 TI - Recent progress in the etiopathogenesis of pediatric biliary disease, particularly Caroli's disease with congenital hepatic fibrosis and biliary atresia. AB - Recent progress in elucidating the etiopathogenesis of pediatric biliary diseases, particularly Caroli's disease with congenital hepatic fibrosis (CHF) and biliary atresia (BA), is reviewed. The former is characterized by multiple saccular dilatations of the intrahepatic bile ducts. An animal model of this disease, the PCK rat, is being extensively studied. PCK rats and Calori's disease with CHF belong to autosomal recessive polycystic kidney disease (ARPKD) with ductal plate malformation. Mutations of PKHD1 have been identified in ARPKD, and fibrocystin, a product of PKHD1 located in the cilia of bile ducts is lacking in the pathologic intrahepatic bile ducts of ARPKD. Disordered cell kinetics, including apoptosis of biliary epithelial cells (BECs), may be significantly related to ductal plate malformation, and laminin and type IV collagen were immunohistochemically reduced in the basement membrane of intrahepatic bile ducts of ARPKD, and such a reduction is an additional factor for the dilatation of bile ducts. Abundant connective tissue growth factor retained diffusely in heparan sulfate proteoglycan in the fibrous portal tracts are responsible for non resolving hepatic fibrosis. In addition, pathologic BECs of ARPKD may acquire mesenchymal features and participate in progressive hepatic fibrosis by producing extracellular matrix molecules. In an animal model of BA, an initial virus induced, T-cell mediated autoimmune-mediated cholangiopathy has been reported. In human BA, virus-induced apoptosis of BECs by a TNF-related apoptosis-inducing ligand followed by the progressive obliteration of bile ducts is also suggested, and epithelial mesenchymal transition of BECs induced by viral infection may be involved in the fibrotic process in sclerosing cholangitis. However, the role of viral infections in the affected tissues is controversial. Comprehensive and analytical studies of ARPKD and BA using human materials and animal models may lead to the clarification of their etiopathogenesis and open the way for new therapeutic strategies. PMID- 20017110 TI - BDNF function and intracellular signaling in neurons. AB - Brain-derived neurotrophic factor (BDNF) and its receptor, TrkB, are broadly expressed in the developing and adult mammalian brain. BDNF/TrkB-stimulated intracellular signaling is critical for neuronal survival, morphogenesis, and plasticity. It is well known that binding of BDNF to TrkB elicits various intracellular signaling pathways, including mitogen-activated protein kinase/extracellular signal-regulated protein kinase (MAPK/ERK), phospholipase Cg (PLCg), and phosphoinositide 3-kinase (PI3K) pathways, and that BDNF exerts biological effects on neurons via activation of similar mechanisms. In addition to TrkB, a low-affinity receptor p75 is also involved in neuronal survival and plasticity. BDNF affects neurons positively or negatively through various intracellular signaling pathways triggered by activation of TrkB or p75. From a clinical standpoint, roles of BDNF have been implicated in the pathophysiology of various brain diseases. The stress-induced steroid hormone, glucocorticoid, and BDNF are putatively associated with the pathophysiology of depression. Recent reports, including our studies, demonstrate possible crosstalk between glucocorticoid- and BDNF/TrkB-mediated signaling. Here, we present a broad overview of the current knowledge concerning BDNF action and associated intracellular signaling as it relates to neuronal protection, synaptic function, and morphological change. Furthermore, understanding the secretion and intracellular dynamics of BDNF proteins is critical as the fate of secreted BDNF may contribute to differences in neuronal response. PMID- 20017111 TI - Contemporary approaches for processing and handling of radical prostactomy specimens. AB - Standardized protocols for processing radical prostatectomy specimens are critical for superior patient management. It provides accurate information to the clinician in a reliable and consistent format to enhance patient care and prognosis. In recent years, processing protocols have been proposed by various authoritative groups, with similar suggestions for most parts of the practice guidelines; however, discrepancy in processing approaches still exists. Standardization improves the quality and consistency of pathology reports. In this review article, we incorporate the processing schemes for radical prostatectomy addressed in literature and propose a comprehensively standardized approach to evaluate radical prostatectomy specimens. PMID- 20017112 TI - An overview on the diversity of cellular organelles during the germ cell cycle. AB - In mammals, germ cells undergo a long journey from specification until sexual maturation. During this journey, which takes place during the entire life cycle of mammals, the germ cells dynamically change their morphology, their expression profile and also the number and character of their cellular bodies. The focus of this review will be the diversity of cellular organelles present in the nucleus and cytoplasm at the different phases of germ cell development. We discuss how these organelles associate and behave to form a multitude of bodies that have long been observed by scientists, and how their presence or absence is used to characterize different stages of germ cell development. These organelles include the female Barr body, polar bodies and Balbiani body; and the male sex body and chromatoid body. It is concluded that compartmentalization of organelles and molecules in the cytoplasm (in particular of mitochondria and RNAs) and of the sex chromosomes in the nucleus seems to be important for regulating germ cell development throughout the life cycle. PMID- 20017113 TI - Application of a stress and coping model to positive and negative adjustment outcomes in colorectal cancer caregiving. AB - This study examined relations between stress and coping predictors and negative (distress) and positive (positive affect, life satisfaction, positive states of mind, health) adjustment outcomes in colorectal cancer caregivers. A total of 622 caregivers and their care-recipients completed questionnaires and interviews. Predictors included caregiving parameters, social support, appraisal (stress, control, challenge), and coping strategies (problem-solving, avoidance, substance use, religion, humor, seeking social support). Regression analyses indicated that after controlling for the effects of relevant caregiving parameters (caregiver age, gender and paid employment status, care-recipient quality of life, cancer stage), better caregiver adjustment was related to higher social support, lower stress and higher challenge appraisals, and less reliance on avoidance and substance use coping, as hypothesized. Stress appraisal emerged as the strongest and most consistent predictor of all adjustment outcomes. Findings support the utility of the stress and coping model of adjustment to caregiving in colorectal cancer. PMID- 20017114 TI - Use of formal and informal mental health resources by cancer survivors: differences between rural and nonrural survivors and a preliminary test of the theory of planned behavior. AB - OBJECTIVE: Previous research has identified rural residence as a risk factor for poorer mental health (MH) outcomes in cancer survivors. This may be due to less use of various MH resources due to poorer access and less favorable attitudes and social norms related to MH resource utilization. The present study sought to examine use of MH resources in rural and nonrural survivors and identify factors associated with MH resource use. METHODS: Cancer survivors (n=113, 1-5 years postdiagnosis) completed a questionnaire packet and telephone interview. Accessibility and postdiagnosis use of various formal and informal MH resources were assessed along with constructs potentially linked to use of MH resources by the Theory of Planned Behavior (TPB; personal attitude, social norm, perceived behavioral control). RESULTS: Results indicated no widespread differences between rural and nonrural cancer survivors in MH resource use although some evidence suggested poorer accessibility and less use of mental health professionals and cancer support groups among rural survivors. In general, rural survivors reported less favorable personal attitudes and social norms regarding MH resource use. TPB constructs accounted for a significant portion of variance in use of most MH resources with personal attitudes generally being the strongest predictor of MH resource use. CONCLUSIONS: Additional research is needed to expand the search for factors, particularly modifiable factors, which might account for disparities in MH outcomes between rural and nonrural survivors. PMID- 20017115 TI - Transitioning childhood cancer survivors to adult-centered healthcare: insights from parents, adolescent, and young adult survivors. AB - OBJECTIVE: To determine Latino adolescent and young adults (AYA) cancer survivors' perceived barriers or facilitators to transition from pediatric to adult-centered survivorship care and to also assess the parents' perspective of care. METHODS: Partnering with a community-based organization that serves Latino survivors, we conducted a qualitative, constant comparative analytic approach exploring in-depth themes that have salience for Latino pediatric cancer survivors seeking care in the adult healthcare setting. Twenty-seven Latino AYA survivors (>or=15 years of age) completed key informant interviews and 21 Latino parents participated in focus groups. RESULTS: Both AYA survivors and parents identified two major facilitative factors for survivorship care: Involvement of the nuclear family in the AYA's survivorship care in the adult healthcare setting and including symptom communication in late effects discussions. Barriers to care included: perceived stigma of a cancer history and continued emotional trauma related to discussions about the childhood cancer experience. CONCLUSIONS: Barriers to survivorship care include cancer stigma for both patient and nuclear family, which can impact on seeking survivorship care due to constraints placed on discussions because it remains difficult to discuss 'cancer' years later. Future research can evaluate if these findings are unique to Latino childhood cancer survivors or are found in other populations of AYA cancer survivors transitioning to adult-centered healthcare. This community-based participatory research collaboration also highlights the opportunity to learn about the needs of childhood cancer survivors from the lens of community leaders serving culturally diverse populations. PMID- 20017116 TI - Grading the commercial optical biosensor literature-Class of 2008: 'The Mighty Binders'. AB - Optical biosensor technology continues to be the method of choice for label-free, real-time interaction analysis. But when it comes to improving the quality of the biosensor literature, education should be fundamental. Of the 1413 articles published in 2008, less than 30% would pass the requirements for high-school chemistry. To teach by example, we spotlight 10 papers that illustrate how to implement the technology properly. Then we grade every paper published in 2008 on a scale from A to F and outline what features make a biosensor article fabulous, middling or abysmal. To help improve the quality of published data, we focus on a few experimental, analysis and presentation mistakes that are alarmingly common. With the literature as a guide, we want to ensure that no user is left behind. PMID- 20017117 TI - Semiblind spatial ICA of fMRI using spatial constraints. AB - Independent component analysis (ICA) utilizing prior information, also called semiblind ICA, has demonstrated considerable promise in the analysis of functional magnetic resonance imaging (fMRI). So far, temporal information about fMRI has been used in temporal ICA or spatial ICA as additional constraints to improve estimation of task-related components. Considering that prior information about spatial patterns is also available, a semiblind spatial ICA algorithm utilizing the spatial information was proposed within the framework of constrained ICA with fixed-point learning. The proposed approach was first tested with synthetic fMRI-like data, and then was applied to real fMRI data from 11 subjects performing a visuomotor task. Three components of interest including two task-related components and the "default mode" component were automatically extracted, and atlas-defined masks were used as the spatial constraints. The default mode network, a set of regions that appear correlated in particular in the absence of tasks or external stimuli and is of increasing interest in fMRI studies, was found to be greatly improved when incorporating spatial prior information. Results from simulation and real fMRI data demonstrate that the proposed algorithm can improve ICA performance compared to a different semiblind ICA algorithm and a standard blind ICA algorithm. PMID- 20017118 TI - Late-postnatal cannabinoid exposure persistently increases FoxP2 expression within zebra finch striatum. AB - Prior work has shown that cannabinoid exposure of zebra finches during sensorimotor stages of vocal development alters song patterns produced in adulthood. We are currently working to identify physiological substrates for this altered song learning. FoxP2 is a transcription factor associated with altered vocal development in both zebra finches and humans. This protein shows a distinct pattern of expression within Area X of striatum that coincides with peak expression of CB(1) cannabinoid receptors during sensorimotor learning. Coincident expression in a brain region essential for song learning led us to test for a potential signaling interaction. We have found that cannabinoid agonists acutely increase expression of FoxP2 throughout striatum. When administered during sensorimotor song learning, cannabinoids increase basal levels of striatal FoxP2 expression in adulthood. Thus, song-altering cannabinoid treatments are associated with persistent increases in basal expression of FoxP2 in zebra finch striatum. PMID- 20017119 TI - Maternal hypoxia increases the activity of MMPs and decreases the expression of TIMPs in the brain of neonatal rats. AB - A recent study has shown that increased activity of matrix metalloproteinases-2 and metalloproteinases-9 (MMP-2 and MMP-9) has detrimental effect on the brain after neonatal hypoxia. The present study determined the effect of maternal hypoxia on neuronal survivability and the activity of MMP-2 and MMP-9, as well as the expression of tissue inhibitors of metalloproteinase 1 and 2 (TIMP-1 and TIMP 2) in the brain of neonatal rats. Pregnant rats were exposed to 10.5% oxygen for 6 days from the gestation day 15 to day 21. Pups were sacrificed at day 0, 4, 7, 14, and 21 after birth. Body weight and brain weight of the pups were measured at each time point. The activity of MMP-2 and MMP-9 and the protein abundance of TIMP-1 and TIMP-2 were determined by zymography and Western blotting, respectively. The tissue distribution of MMPs was examined by immunofluorescence staining. The neuronal death was detected by Nissl staining. Maternal hypoxia caused significant decreases in body and brain size, increased activity of MMP-2 at day 0, and increased MMP-9 at day 0 and 4. The increased activity of the MMPs was accompanied by an overall tendency towards a reduced expression of TIMPs at all ages with the significance observed for TIMPs at day 0, 4, and 7. Immunofluorescence analysis showed an increased expression of MMP-2, MMP-9 in the hippocampus at day 0 and 4. Nissl staining revealed significant cell death in the hippocampus at day 0, 4, and 7. Functional tests showed worse neurobehavioral outcomes in the hypoxic animals. PMID- 20017121 TI - Systematic LC-MS/MS bioanalytical method development that incorporates plasma phospholipids risk avoidance, usage of incurred sample and well thought-out chromatography. AB - This treatise summarizes the underlying principle and the road map for systematic LC-MS/MS bioanalytical method development. The three themes that have recently emerged as central to building quality during method development-phospholipids, incurred sample and sound chromatographic considerations-are the main focus of this article. In order to reduce the bioanalytical risk associated with plasma phospholipids, a dual approach involving extraction and chromatography is recommended. The use of incurred sample during method development is essential to avoid interference arising from drug-related components. This is different from the current practice of incurred sample reanalysis, which tests reproducibility during method application. LC column/mobile phase optimization is needed to achieve appropriate peak shape, sensitivity and the separation of the analyte from interfering metabolites and phospholipids. Related to sound chromatographic considerations, we lay out facts and myths related to UPLC, vis-a-vis HPLC. Incorporation of quality during method development avoids the costly experience of finding out by chance about the invalidity of a method after it has been used in support of a number of pivotal clinical and non-clinical studies. To this end, we put forth an outline of a protocol for a systematic LC-MS/MS bioanalytical method development. PMID- 20017120 TI - Analysis of endogenous glutathione-adducts and their metabolites. AB - The ability to conduct validated analyses of glutathione (GSH)-adducts and their metabolites is critically important in order to establish whether they play a role in cellular biochemical or pathophysiological processes. The use of stable isotope dilution (SID) methodology in combination with liquid chromatography tandem mass spectrometry (LC-MS/MS) provides the highest bioanalytical specificity possible for such analyses. Quantitative studies normally require the high sensitivity that can be obtained by the use of multiple reaction monitoring (MRM)/MS rather than the much less sensitive but more specific full scanning methodology. The method employs a parent ion corresponding to the intact molecule together with a prominent product ion that obtained by collision induced dissociation. Using SID LC-MRM/MS, analytes must have the same relative LC retention time to the heavy isotope internal standard established during the validation procedure, the correct parent ion and the correct product ion. This level of specificity cannot be attained with any other bioanalytical technique employed for biomarker analysis. This review will describe the application of SID LC-MR/MS methodology for the analysis of GSH-adducts and their metabolites. It will also discuss potential future directions for the use of this methodology for rigorous determination of their utility as disease and exposure biomarkers. PMID- 20017122 TI - Dried blood spot sampling in combination with LC-MS/MS for quantitative analysis of small molecules. AB - The collection of whole blood samples on paper, known as dried blood spot (DBS), dates back to the early 1960s in newborn screening for inherited metabolic disorders. DBS offers a number of advantages over conventional blood collection. As a less invasive sampling method, DBS offers simpler sample collection and storage and easier transfer, with reduced infection risk of various pathogens, and requires a smaller blood volume. To date, DBS-LC-MS/MS has emerged as an important method for quantitative analysis of small molecules. Despite the increasing popularity of DBS-LC-MS/MS, the method has its limitations in assay sensitivity due to the small sample size. Sample quality is often a concern. Systematic assessment on the potential impact of various blood sample properties on accurate quantification of analyte of interest is necessary. Whereas most analytes may be stable on DBS, unstable compounds present another challenge for DBS as enzyme inhibitors cannot be conveniently mixed during sample collection. Improvements on the chemistry of DBS card are desirable. In addition to capturing many representative DBS-LS-MS/MS applications, this review highlights some important aspects of developing and validating a rugged DBS-LC-MS/MS method for quantitative analysis of small molecules along with DBS sample collection, processing and storage. PMID- 20017124 TI - Fast determination of the optimal rotational matrix for macromolecular superpositions. AB - Finding the rotational matrix that minimizes the sum of squared deviations between two vectors is an important problem in bioinformatics and crystallography. Traditional algorithms involve the inversion or decomposition of a 3 x 3 or 4 x 4 matrix, which can be computationally expensive and numerically unstable in certain cases. Here, we present a simple and robust algorithm to rapidly determine the optimal rotation using a Newton-Raphson quaternion-based method and an adjoint matrix. Our method is at least an order of magnitude more efficient than conventional inversion/decomposition methods, and it should be particularly useful for high-throughput analyses of molecular conformations. PMID- 20017125 TI - A new unconventional halogen bond C-X...H-M between HCCX (X = Cl and Br) and HMH (M = Be and Mg): an ab initio study. AB - In this article, a new type of halogen-bonded complex YCCX...HMY (X = Cl, Br; M = Be, Mg; Y = H, F, CH(3)) has been predicted and characterized at the MP2/aug-cc pVTZ level. We named it as halogen-hydride halogen bonding. In each YCCX...HMY complex, a halogen bond is formed between the positively charged X atom and the negatively charged H atom. This new kind of halogen bond has similar characteristics to the conventional halogen bond, such as the elongation of the C X bond and the red shift of the C-X stretch frequency upon complexation. The interaction strength of this type of halogen bond is in a range of 3.34-10.52 kJ/mol, which is smaller than that of dihydrogen bond and conventional halogen bond. The nature of the electrostatic interaction in this type of halogen bond has also been unveiled by means of the natural bond orbital, atoms in molecules, and energy decomposition analyses. PMID- 20017126 TI - Selective oxidation of styrene on an oxygen-adsorbed Cu(111): a comparison with Au(111). AB - The reaction mechanism for the styrene selective oxidation on the oxygen preadsorbed Cu(111) surface has been studied by the density functional theory calculation with the periodic slab model. The calculated result indicated that the process includes two steps: forming the oxametallacycle intermediate (OMMS) and then producing the products. In addition, it was found that the second step, from OMMS to the product, is the rate-controlling step, which is similar to the previous work of ethylene selective oxidation. The present result indicated that the selectivity towards the formation of styrene epoxide on Cu(111) is much higher than that on Au(111). More importantly, we found that the mechanism via the OMMS (2) (i.e., the preadsorbed atomic oxygen bound to the CH(2) group involved in C(6)H(5)-CH=CH(2)) to produce styrene epoxide is kinetically favored than that of OMMS (1). We also found that the selectivity toward the styrene epoxide formation on Cu(2)O is similar to that of Cu(111). PMID- 20017127 TI - A rapid and simple chemiluminescence method for screening levels of inosine and hypoxanthine in non-traumatic chest pain patients. AB - A rapid and simple chemiluminescence method was developed for detection of inosine and hypoxanthine in human plasma. The method utilized a microplate luminometer with direct injectors to automatically dispense reagents during sample analysis. Enzymatic conversions of inosine to hypoxanthine, followed by hypoxanthine to xanthine to uric acid, generated superoxide anion radicals as a useful metabolic by-product. The free radicals react with Pholasin((r)) , a sensitive photoprotein used for chemiluminescence detection, to produce measurable blue-green light. The use of Pholasin((r)) and a chemiluminescence signal enhancer, Adjuvant-KTM, eliminated the need for plasma clean-up steps prior to analysis. The method used 20 MUL of heparinized plasma, with complete analysis of total hypoxanthine levels (inosine is metabolized to hypoxanthine using purine nucleoside phosphorylase) in approximately 3.7 min. The rapid chemiluminescence method demonstrated the capability of differentiating total hypoxanthine levels between healthy individuals, and patients presenting with non traumatic chest pain and potential acute cardiac ischemia. The results support the potential use of chemiluminescence methodology as a diagnostic tool to rapidly screen for elevated levels of inosine and hypoxanthine in human plasma, potential biomarkers of acute cardiac ischemia. PMID- 20017128 TI - Sensitive determination of epinephrine in pharmaceutical preparation by flow injection coupled with chemiluminescence detection and mechanism study. AB - A novel, rapid and sensitive method was described for the determination of epinephrine (EP) using flow injection analysis coupled with chemiluminescence (CL) detection, which based on EP enhanced the weak CL emission of luminol-KIO(4) system in NaOH solution. Parameters affecting the CL intensity and reproducibility were optimized systematically. Under the optimized experiment conditions, the net CL intensity was proportional to the concentration of EP in the range of 5.0 * 10(-8) to 1.5 * 10(-6) mol/L with a detection limit of 1.9 * 10(-9) mol/L. The relative standard deviation (RSD) was found to be 0.7% for 13 replicate determinations of 3.0 * 10(-7) mol/L EP. The applicability of the proposed method was illustrated in the determination of EP in pharmaceutical preparation. The recoveries of EP at different levels in EP hydrochloride injection were between 95.4 and 104.7%. One assay procedure takes only 27 s, and the sampling rate was calculated about to be 130 samples/h. The possible mechanism of the enhanced CL intensity was studied by examining CL spectra and UV vis spectra. PMID- 20017129 TI - Effect of concentrating and exposing the bioluminescent bacteria to the non luminescent allo-bacterial extracellular products on their luminescence. AB - Bioluminescence is a biochemical process occurring in many organisms. Bacterial bioluminescence has been investigated extensively that lead to many applications of such knowledge. Quorum sensing in the bioluminescent bacteria is a chemical signal process to recognize the strength of its own population to start luminescence in harmony. There is a mechanism in these bacteria to also recognize inter-species strength. When there is a higher number of these bacteria, the possibility and frequency of cell-cell physical contact will be high. In this study, the physical proximity was artificially enhanced between cells and the effect on luminescence in the concentrated cells in the normal culture medium and in the presence of other non-bacterial cell-free supernatants was investigated. The role of such physical contact in the quorum sensing in the bioluminescence is not known. Increase in the luminescence of V. fischeri when concentrated shows that the presence of physical proximity facilitates the quorum sensing for their bioluminescence. PMID- 20017130 TI - L-Cysteine capped CdTe-CdS core-shell quantum dots: preparation, characterization and immuno-labeling of HeLa cells. AB - Functionalized CdTe-CdS core-shell quantum dots (QDs) were synthesized in aqueous solution via water-bathing combined hydrothermal method using L-cysteine (L-Cys) as a stabilizer. This method possesses both the advantages of water-bathing and hydrothermal methods for preparing high-quality QDs with markedly reduced synthesis time, and better stability than a lone hydrothermal method. The QDs were characterized by transmission electronic microscopy and powder X-ray diffraction and X-ray photoelectron spectroscopy. The CdTe-CdS QDs with core shell structure showed both enhanced fluorescence and better photo stability than nude CdTe QDs. After conjugating with antibody rabbit anti-CEACAM8 (CD67), the as prepared l-Cys capped CdTe-CdS QDs were successfully used as fluorescent probes for the direct immuno-labeling and imaging of HeLa cells. It was indicated that this kind of QD would have application potential in bio-labeling and cell imaging. PMID- 20017131 TI - Synthesis and luminescence properties of Zn(Cu(0.01) Cd(0.02) Mg(0.02))S phosphor. AB - To improve the luminescence properties of ZnS : Cu, a multi-doped method was used to prepare Zn(Cu(0.01) Cd(0.02) Mg(0.02))S in this paper. As a new designed semiconductor compound, particular properties of Zn(Cu(0.01) Cd(0.02) Mg(0.02))S were investigated, especially luminescence intensity and lifetime. Structure and compositions of Zn(Cu(0.01) Cd(0.02) Mg(0.02))S phosphor were analyzed by XRD and TEM respectively. Luminescence intensity and luminescence lifetime of the prepared Zn(Cu(0.01) Cd(0.02) Mg(0.02))S phosphor were investigated by luminescence spectrometry. As a result, compared with pure ZnS or doped ZnS phosphors, Zn(Cu(0.01) Cd(0.02) Mg(0.02))S showed remarkably improved luminescence properties. The optimum ratio of three dopants was obtained for solving luminescence problems. PMID- 20017133 TI - The SRI24 multichannel atlas of normal adult human brain structure. AB - This article describes the SRI24 atlas, a new standard reference system of normal human brain anatomy, that was created using template-free population registration of high-resolution magnetic resonance images acquired at 3T in a group of 24 normal control subjects. The atlas comprises anatomical channels (T1, T2, and proton density weighted), diffusion-related channels (fractional anisotropy, mean diffusivity, longitudinal diffusivity, mean diffusion-weighted image), tissue channels (CSF probability, gray matter probability, white matter probability, tissue labels), and two cortical parcellation maps. The SRI24 atlas enables multichannel atlas-to-subject image registration. It is uniquely versatile in that it is equally suited for the two fundamentally different atlas applications: label propagation and spatial normalization. Label propagation, herein demonstrated using diffusion tensor image fiber tracking, is enabled by the increased sharpness of the SRI24 atlas compared with other available atlases. Spatial normalization, herein demonstrated using data from a young-old group comparison study, is enabled by its unbiased average population shape property. For both propagation and normalization, we also report the results of quantitative comparisons with seven other published atlases: Colin27, MNI152, ICBM452 (warp5 and air12), and LPBA40 (SPM5, FLIRT, AIR). Our results suggest that the SRI24 atlas, although based on 3T MR data, allows equally accurate spatial normalization of data acquired at 1.5T as the comparison atlases, all of which are based on 1.5T data. Furthermore, the SRI24 atlas is as suitable for label propagation as the comparison atlases and detailed enough to allow delineation of anatomical structures for this purpose directly in the atlas. PMID- 20017134 TI - Amygdala damage affects event-related potentials for fearful faces at specific time windows. AB - The amygdala is known to influence processing of threat-related stimuli in distant brain regions, including visual cortex. The time-course of these distant influences is unknown, although this information is important for resolving debates over likely pathways mediating an apparent rapidity in emotional processing. To address this, we recorded event-related potentials (ERPs) to seen fearful face expressions, in preoperative patients with medial temporal lobe epilepsy who had varying degrees of amygdala pathology, plus healthy volunteers. We found that amygdala damage diminished ERPs for fearful versus neutral faces within the P1 time-range, approximately 100-150 ms, and for a later component at approximately 500-600 ms. Individual severity of amygdala damage determined the magnitude of both these effects, consistent with a causal amygdala role. By contrast, amygdala damage did not affect explicit perception of fearful expressions nor a distinct emotional ERP effect at 150-250 ms. These results demonstrate two distinct time-points at which the amygdala influences fear processing. The data also demonstrate that while not all aspects of expression processing are disrupted by amygdala damage, there is a crucial impact on an early P1 component. These findings are consistent with the existence of multiple processing stages or routes for fearful faces that vary in their dependence on amygdala function. PMID- 20017135 TI - Determination of side-chain-rotamer and side-chain and backbone virtual-bond stretching potentials of mean force from AM1 energy surfaces of terminally blocked amino-acid residues, for coarse-grained simulations of protein structure and folding. II. Results, comparison with statistical potentials, and implementation in the UNRES force field. AB - Using the harmonic-approximation approach of the accompanying article and AM1 energy surfaces of terminally blocked amino-acid residues, we determined physics based side-chain rotamer potentials and the side-chain virtual-bond-deformation potentials of 19 natural amino-acid residues with side chains. The potentials were approximated by analytical formulas and implemented in the UNRES mesoscopic dynamics program. For comparison, the corresponding statistical potentials were determined from 19,682 high-resolution protein structures. The low free-energy region of both the AM1-derived and the statistical potentials is determined by the valence geometry and the L-chirality, and its size increases with side-chain flexibility and decreases with increasing virtual-bond-angle theta. The differences between the free energies of rotamers are greater for the AM1-derived potentials compared with the statistical potentials and, for alanine and other residues with small side chains, a region corresponding to the C(ax)(7) conformation has remarkably low free-energy for the AM1-derived potentials, as opposed to the statistical potentials. These differences probably result from the interactions between neighboring residues and indicate the need for introduction of cooperative terms accounting for the coupling between side-chain rotamer and backbone interactions. Both AM1-derived and statistical virtual-bond-deformation potentials are multimodal for flexible side chains and are topologically similar; however, the regions of minima of the statistical potentials are much narrower, which probably results from imposing restraints in structure determination. The force field with the new potentials was preliminarily optimized using the FBP WW domain (1E0L) and the engrailed homeodomain (1ENH) as training proteins and assessed to be reasonably transferable. PMID- 20017136 TI - Isoalvaxanthone inhibits colon cancer cell proliferation, migration and invasion through inactivating Rac1 and AP-1. AB - Isoalvaxanthone (IAX) is a bioactive xanthone isolated from Cudrania cochinchinensis (Lour.). However, the function and mechanism of this compound in cancer migration and invasion have not been elucidated to date. In this study, we found that IAX could suppress various steps of tumor metastasis including proliferation, migration and invasion in a dose-dependent manner on colorectal cancer cells. Especially matrix metalloproteinase 2 (MMP-2), the pivotal factor in cancer invasion, was suppressed both on activation and expression after treated with IAX. To understand the underlying mechanism of IAX on the inhibitory effect of proliferation, migration and invasion, we demonstrated that IAX could significantly inhibit the activation of Rac1 but has undetectable effect on GTP RhoA, GTP-Cdc42 and the phosphorylation of ERK1/2, p38 MAPK and JNK. Moreover, IAX showed little influence on the transcriptional activity of nuclear transcription factor kappaB (NF-kappaB) but strongly inhibited that of activator protein-1 (AP-1), which is the downstream transcriptional factor of Rac1. Together, our results indicate that IAX exerts anticancer effect in SW620 cells by targeting MMP-2 via regulating the activity of Rac1 and AP-1. These results are the first to reveal the function of IAX in tumor metastasis and its underlying molecular mechanism, thus suggest IAX to be a promising antimetastatic agent. PMID- 20017137 TI - Cooperation of tumor-derived HBx mutants and p53-249(ser) mutant in regulating cell proliferation, anchorage-independent growth and aneuploidy in a telomerase immortalized normal human hepatocyte-derived cell line. AB - Hepatocellular carcinoma (HCC) is a common cancer, and hepatitis B virus (HBV) is a major etiological agent. Convincing epidemiological and experimental evidence also links HCC to aflatoxin, a naturally occurring mycotoxin that produces a signature p53-249(ser) mutation. Recently, we have reported that tumor-derived HBx variants encoded by HBV exhibited attenuated transactivation and proapoptotic functions but retained their ability to block p53-mediated apoptosis. These results indicate that mutations in HBx may contribute to the development of HCC. In this study, we determined whether tumor-derived HBx mutants along, or in cooperation with p53-249(ser), could alter cell proliferation and chromosome stability of normal human hepatocytes. To test this hypothesis, we established a telomerase immortalized normal human hepatocycte line HHT4 that exhibited a near diploid karyotype and expressed many hepatocyte-specific genes. We found that overexpression one of the tumor-derived HBx mutants, CT, significantly increased colony forming efficiency (CFE) while its corresponding wild-type allele CNT significantly decreased CFE in HHT4 cells. p53-249(ser) rescued CNT-mediated inhibition of colony formation. Although HHT4 cells lacked an anchorage independent growth capability as they did not form any colonies in soft agar, the CT-expressing HHT4 cells could form colonies, which could be significantly enhanced by p53-249(ser). Induction of aneuploidy could be observed in HHT4 cells expressing CT, but additionally recurring chromosome abnormalities could only be detected in cells coexpressing CT and p53-249(ser). Our results are consistent with the hypothesis that certain mutations in HBx and p53 at codon 249 may cooperate in contributing to liver carcinogenesis. PMID- 20017138 TI - Core 2 N-acetylglucosaminyltransferase-1 expression induces aggressive potential of testicular germ cell tumor. AB - We studied orchiectomy specimens from 130 patients immuhistochemically with testicular germ cell tumor (TGCT) using anti-core 2 N acetylglucosaminyltransferase-1 (C2GnT-1) antibody. The incidence of C2GnT-1 positivity in stage I disease (29.5%, 21/71) was significantly lower than that in higher stages (84.7%, 50/59) (P < 0.001, chi(2) test). This significant difference was also found when the cases were divided into seminoma and NSGCT according to histopathological classification. Kaplan-Meier plots and the log rank test showed that in the patients with stage I seminoma, C2GnT-1-positive cases had a higher risk for recurrence (P < 0.001). This was also the case with the patients with stage I NSGCT (P < 0.001). To determine whether C2GnT-1 promotes aggressive behavior of cancer cells, a C2GnT-1-negative human TGCT cell line, JKT-1, was stably transfected with a mammalian expression vector containing C2GnT-1 cDNA. In vitro assays revealed that JKT-1-C2 cells are more invasive than mock transfectants, although there are no differences in proliferation activity. When orthotopically inoculated into athymic nude mice, JKT-1-C2 cells produced larger testicular tumors extending to the retroperitoneum with mesenteric metastasis, while mock transfectants produced small tumors without metastasis (P < 0.01, Mann-Whitney's U-test). When injected via the tail vein, JKT-1-C2 cells produced a number of metastatic lung foci. In contrast, mock transfectants produced a small number of nodules (p < 0.01, Mann-Whitney's U-test). These results strongly suggest that C2GnT-1 enhances the metastatic potential of TGCT and may be a reliable biomarker for aggressive potential of TGCT. PMID- 20017139 TI - MicroRNA miR-146a is induced by HTLV-1 tax and increases the growth of HTLV-1 infected T-cells. AB - Human T-cell leukemia virus type 1 (HTLV-1) is the causative agent of adult T cell leukemia (ATL), which is an aggressive and fatal CD4(+) T cell malignancy. MicroRNA (miRNA), a novel class of RNA that regulates gene expression, is involved in many cellular processes such as growth, development and apoptosis. It has recently been linked to several cancer phenotypes. However, aberrant miRNA expression and its pathologic significance in ATL are not well documented. Here, we investigated the role of miRNAs in HTLV-1-related leukemogenesis. The results showed that miR-146a was upregulated in HTLV-1-infected T-cell lines compared to uninfected T-cell lines. Tax-induced miR-146a expression in a NF-kappaB-dependent manner and inhibited the expression of gene harboring the target sequence of miR 146a on its 3'UTR. Inhibition of miR-146a function by anti-miRNA inhibitor reduced the proliferation of HTLV-1-infected T-cell lines but not that of uninfected T-cell lines. Moreover, overexpression of miR-146a enhanced the growth of an HTLV-1-infected T-cell line. Our findings suggest that miR-146a is a potentially suitable therapeutic target of ATL. PMID- 20017140 TI - Evidence of heterogeneity within colorectal liver metastases for allelic losses, mRNA level expression and in vitro response to chemotherapeutic agents. AB - A goal of oncology is to predict chemosensitivity of tumors. This approach assumes that in a patient all tumor deposits are homogeneous. We have tested the heterogeneity between several samples of the same liver metastasis (LM; intrametastatic heterogeneity) or between multiple LM (intermetastatic heterogeneity) from colorectal cancer in a single patient. In 16 untreated patients, several fragments of LM and nontumorous liver were collected. Heterogeneity to anticancer drug treatment was assessed in vitro on primary tissue cultures on poly-HEMA-coated surface with or without the topoisomerase-I inhibitor metabolite SN-38. Heterogeneity of response to SN-38 was observed in 55% of cases from one fragment to another in the same LM and in 64% of cases from one LM to another in the same patient. Allelic losses were characterized on 5q, 8p, 17p, 18q, 22q using 29 microsatellites markers. Seven patients (58%) had a perfect homogeneity for allelic losses in their LM whereas 3 (21%) had intrametastatic and 2 (18%) had intermetastatic heterogeneity. The analysis of gene expression was carried out by real time RT-PCR quantification using specific probes for TS, TOPO1, ERCC1, and CES2. Level expression of genes tested appeared heterogeneous with average variations of 57(+ or - 23)%, 52(+ or - 18)%, 53(+ or 18)%, 56(+ or - 16)% for TS, TOPO1, ERCC1, and CES2 respectively for intermetastatic variability and 47(+ or - 26)%, 36(+ or - 14)%, 38(+ or - 19)%, and 56(+ or - 29)%, respectively for intrametastatic variability. Our results demonstrate intermetastatic and intrametastatic heterogeneity suggesting that pretherapeutic analysis of a single tumor biopsy is likely to lead to a misinterpretation of sensitivity to anticancer treatment. PMID- 20017141 TI - Synthesis and thermal expansion of copper nanotubes and nanowires with Y- and step-shaped topologies. PMID- 20017142 TI - Low-density lipoprotein (LDL) receptor mRNA and protein may enable LDL to replace bovine serum albumin during the in vitro swine embryo development. PMID- 20017144 TI - Proteomic analysis of doxorubicin-induced changes in the proteome of HepG2cells combining 2-D DIGE and LC-MS/MS approaches. AB - HepG-2 cells are widely used as a cell model to investigate hepatocellular carcinomas and the effect of anticancer drugs such as doxorubicin, an effective antineoplastic agent, which has broad antitumoral activity against many solid and hematological malignancies. To investigate the effect of doxorubicin on the protein pattern, we used complementary proteomic workflows including 2-D gel based and gel-free methods. The analysis of crude HepG2 cell extracts by 2-D DIGE provided data on 1835 protein spots which was then complemented by MS-centered analysis of stable isotope labeling by amino acids in cell culture-labeled cells. The monitoring of more than 1300 distinct proteins, including proteins of the membrane fraction provides the most comprehensive overview on the proteome of the widely used model cell line HepG2. Of the proteins monitored in total, 155 displayed doxorubicin-induced changes in abundance. Functional analysis revealed major influences of doxorubicin on proteins involved in protein synthesis, DNA damage control, electron transport/mitochondrial function, and tumor growth. The strongest decrease in level was found for proteins involved in DNA replication and protein synthesis, whereas proteins with a function in DNA damage control and oxidative stress management displayed increased levels following treatment with doxorubicin compared with control cells. Furthermore, the doxorubicin-associated increase in levels of multiple forms of keratins 8, 18, and 19 and other structural proteins revealed an influence on the cytoskeleton network. PMID- 20017143 TI - Sperm-borne protein, PAWP, initiates zygotic development in Xenopus laevis by eliciting intracellular calcium release. AB - We previously reported postacrosomal sheath WW domain binding protein (PAWP) as a candidate sperm borne, oocyte-activating factor. PAWP enters the oocyte during fertilization and induces oocyte activation events including meiotic resumption, pronuclear formation, and egg cleavage. However, in order to provide proof that PAWP is a primary initiator of zygotic development it is imperative to show that PAWP initiates intracellular calcium signaling, which is considered essential for oocyte activation. Utilizing Xenopus laevis as our model, we injected recombinant PAWP or Xenopus sperm into metaphase II-arrested oocytes and observed a significant rise in intracellular calcium levels over controls. Concurring intensities and durations of PAWP and sperm-induced calcium waves, detected by infrared two-photon laser-scanning fluorescence microscopy, were prevented by coinjection of a competitive PPGY-containing peptide derived from PAWP but not by the point-mutated form of this peptide. This study also correlates PAWP and sperm induced calcium release with meiotic resumption in Xenopus. The similar mode of oocyte activation, and the ability of the competitive peptide in blocking both sperm- and PAWP-induced calcium release, provide evidence for the first time that sperm-anchored PAWP is a primary initiator of zygotic development. PMID- 20017145 TI - Proteomic analysis of the phytopathogenic soilborne fungus Verticillium dahliae reveals differential protein expression in isolates that differ in aggressiveness. AB - Verticillium dahliae is a soilborne fungus that causes a vascular wilt disease of plants and losses in a broad range of economically important crops worldwide. In this study, we compared the proteomes of highly (Vd1396-9) and weakly (Vs06-14) aggressive isolates of V. dahliae to identify protein factors that may contribute to pathogenicity. Twenty-five protein spots were consistently observed as differential in the proteome profiles of the two isolates. The protein sequences in the spots were identified by LC-ESI-MS/MS and MASCOT database searches. Some of the identified sequences shared homology with fungal proteins that have roles in stress response, colonization, melanin biosynthesis, microsclerotia formation, antibiotic resistance, and fungal penetration. These are important functions for infection of the host and survival of the pathogen in soil. One protein found only in the highly aggressive isolate was identified as isochorismatase hydrolase, a potential plant-defense suppressor. This enzyme may inhibit the production of salicylic acid, which is important for plant defense response signaling. Other sequences corresponding to potential pathogenicity factors were identified in the highly aggressive isolate. This work indicates that, in combination with functional genomics, proteomics-based analyses can provide additional insights into pathogenesis and potential management strategies for this disease. PMID- 20017147 TI - A celebration of protein-protein interactions. PMID- 20017148 TI - Glucose-regulated protein 78: a new partner of p53 in trophoblast. AB - Although wild-type p53 protein is overexpressed in first trimester trophoblast, it is inactive towards its target genes Metalloproteinase 2 and 9. This seems to be due to a complex mechanism of inactivation and stabilization of p53 relying on the formation of protein complexes involving the N-terminus of p53. To detect the proteins associated with this sequence, we incubated biotinylated p53 N-terminal peptide in cytotrophoblastic cell medium 24 h before lysis of cells. We purified the proteins retained on biotinylated peptide using a neutravidin affinity column. Proteins were then identified by peptide mass finger printing followed or not by peptide fragmentation sequencing. Among these proteins, we identified glucose-regulated protein 78 (GRP78) and verified its interaction with p53 in trophoblastic cells by immunoprecipitation and Western blot analysis. Moreover, the decreased expression of GRP78 induced by GRP78siRNA or versipelostatin decreased the formation of high molecular weight p53 complexes and p53 monomer and increased trophoblastic invasion. These results suggest that GRP78 is involved in inactivation and stabilization of p53 and in the regulation of trophoblastic invasion. PMID- 20017149 TI - Visions: the art of science. PMID- 20017154 TI - Molecular characteristics and physiological functions of major royal jelly protein 1 oligomer. AB - Royal jelly contains numerous components, including proteins. Major royal jelly protein (MRJP) 1 is the most abundant protein among the soluble royal jelly proteins. In its physiological state, MRJP 1 exists as a monomer and/or oligomer. This study focuses the molecular characteristics and functions of MRJP 1 oligomer. MRJP 1 oligomer purified using HPLC techniques was subjected to the following analyses. The molecular weight of MRJP 1 oligomer was found to be 290 kDa using blue native-PAGE. MRJP 1 oligomer was separated into 55 and 5 kDa spots on 2-D blue native/SDS-PAGE. The 55 kDa protein was identified as MRJP 1 monomer by proteome analysis, whereas the 5 kDa protein was identified as Apisimin by N terminal amino acid sequencing, and this protein may function as a subunit joining protein within MRJP 1 oligomer. We also found that the oligomeric form included noncovalent bonds and was stable under heat treatment at 56 degrees C. Furthermore, MRJP 1 oligomer dose dependently enhanced and sustained cell proliferation in the human lymphoid cell line Jurkat. In conclusion, MRJP 1 oligomer is a heat-resistant protein comprising MRJP 1 monomer and Apisimin, and has cell proliferation activity. These findings will contribute to further studies analyzing the effects of MRJP 1 in humans. PMID- 20017155 TI - Protein profiling in human sera for identification of potential lung cancer biomarkers using antibody microarray. AB - To identify potential biomarkers of lung cancer (LC), profiling of proteins in sera obtained from healthy and LC patients was determined using an antibody microarray. Based on our previous study on mRNA expression profiles between patients with LC and healthy persons, 19 proteins of interest were selected as targets for fabrication of an antibody microarray. Antibody to each protein and five nonspecific control antibodies were spotted onto a hydrogel-coated glass slide and used for profiling of proteins in sera of LC patients in a two-color fluorescence assay. Forty-eight human sera samples were analyzed, and expression profiling of proteins were represented by the internally normalized ratio method. Six proteins were distinctly down-regulated in sera of LC patients; this observation was validated by Wilcoxon test, false discovery rate, and Western blotting. Blind test of other 32 human sera using the antibody microarray followed by hierarchical clustering analysis revealed an approximate sensitivity of 88%, specificity of 80%, and an accuracy of 84%, respectively, in classifying the sera, which supports the potential of the six identified proteins as biomarkers for the prognosis of lung cancer. PMID- 20017156 TI - Membrane protein analysis using an improved peptic in-solution digestion protocol. AB - In the proteomic analysis of membrane proteins, less-specific proteases have become a promising tool to overcome fundamental limitations of trypsin with its unique specificity for basic residues. Pepsin is well-known to be utilized for specific applications that require acidic conditions, but in terms of membrane protein identification and characterization, it has been disregarded for the most part. This work presents an optimization of an existing peptic digest protocol for the analysis of membrane proteins using bacteriorhodopsin from purple membranes as reference. PMID- 20017157 TI - Liver tumor ablation: percutaneous and open approaches. AB - The global incidence of liver cancer is greater than a million cases a year. Surgical resection where applicable is still the standard of care for these patients. Various liver-directed regional therapies have been developed in an effort to treat the vast majority of unresectable liver tumors. This article reviews the principles behind various ablation therapies currently available for malignant liver tumors and their outcomes. PMID- 20017158 TI - Hepatic ablation for neuroendocrine tumor metastases. AB - The indolent nature of neuroendocrine tumors and their proclivity to be hormonally active warrant aggressive treatment for advanced stage disease with hepatic metastases. Cytoreduction has been associated with improved symptom control as well as prolonged survival compared with those treated with medical therapy. The primary modalities of cytoreduction employed are resection, ablation, and embolization. In particular, radiofrequency ablation has been utilized with good results and minimal morbidity for treating patients with advanced neuroendocrine disease. PMID- 20017159 TI - Renal ablative therapy: radiofrequency ablation and cryoablation. AB - Widespread use of cross-sectional imaging has contributed to an increase in diagnosis of small renal masses. While extirpation has historically been the "gold standard" for managing such tumors, thermal ablation is increasingly utilized as primary surgical therapy. Contemporary series suggest that ablative procedures maintain oncologic outcomes similar to extirpation while minimizing patient morbidity. As more renal masses are diagnosed in elderly or comorbid patients, ablation will likely assume a central role in management strategies. PMID- 20017160 TI - 3D compressed sensing for highly accelerated hyperpolarized (13)C MRSI with in vivo applications to transgenic mouse models of cancer. AB - High polarization of nuclear spins in liquid state through hyperpolarized technology utilizing dynamic nuclear polarization has enabled the direct monitoring of (13)C metabolites in vivo at a high signal-to-noise ratio. Acquisition time limitations due to T(1) decay of the hyperpolarized signal require accelerated imaging methods, such as compressed sensing, for optimal speed and spatial coverage. In this paper, the design and testing of a new echo planar (13)C three-dimensional magnetic resonance spectroscopic imaging (MRSI) compressed sensing sequence is presented. The sequence provides up to a factor of 7.53 in acceleration with minimal reconstruction artifacts. The key to the design is employing x and y gradient blips during a fly-back readout to pseudorandomly undersample k(f)-k(x)-k(y) space. The design was validated in simulations and phantom experiments where the limits of undersampling and the effects of noise on the compressed sensing nonlinear reconstruction were tested. Finally, this new pulse sequence was applied in vivo in preclinical studies involving transgenic prostate cancer and transgenic liver cancer murine models to obtain much higher spatial and temporal resolution than possible with conventional echo-planar spectroscopic imaging methods. PMID- 20017161 TI - Parallel excitation in the human brain at 9.4 T counteracting k-space errors with RF pulse design. AB - Multidimensional spatially selective radiofrequency (RF) pulses have been proposed as a method to mitigate transmit B1 inhomogeneity in MR experiments. These RF pulses, however, have been considered impractical for many years because they typically require very long RF pulse durations. The recent development of parallel excitation techniques makes it possible to design multidimensional RF pulses that are short enough for use in actual experiments. However, hardware and experimental imperfections can still severely alter the excitation patterns obtained with these accelerated pulses. In this note, we report at 9.4 T on a human eight-channel transmit system, substantial improvements in two-dimensional excitation pattern accuracy obtained when measuring k-space trajectories prior to parallel transmit RF pulse design (acceleration x4). Excitation patterns based on numerical simulations closely reproducing the experimental conditions were in good agreement with the experimental results. PMID- 20017162 TI - Radiofrequency and microwave ablation of lung tumors. AB - Lung cancer is the most common cause of cancer-related mortality in the United States. Surgical resection remains the standard treatment for early stage, non small cell lung cancer, and limited pulmonary metastases. However, many patients with resectable disease have significant co-morbidities which preclude surgical resection. Radiofrequency and microwave ablation are emerging modalities of treatment for these high-risk patients. Here, we review the principles, common devices in use, and treatment results of ablative therapy for lung tumors. PMID- 20017163 TI - Radial-template approach for accurate density representation in computational quantum theory. AB - A method for improved representation of electronic charge and spin densities for molecular and solid state systems is presented, based upon extensions of least squares fits to quantum mechanical "true" densities using basis functions of limited support. Attention is given to optimization of radial degrees of fit freedom, and the design of fit functions permitting rapid analytic manipulation and calculation of properties, such as Coulomb potentials. The method is demonstrated for covalent CO and for a large metal-organic crystalline structure. PMID- 20017164 TI - POU5F1P1, a putative cancer susceptibility gene, is overexpressed in prostatic carcinoma. AB - BACKGROUND: Association between genetic variants located on human chromosome 8q24.21 with an increased risk for prostatic carcinoma has been well established. POU5F1P1, a processed pseudogene homologous to the pluripotency factor OCT4, is the only sequence with coding capacity in this region. The objective of this study was to investigate the POU5F1P1 expression in prostatic carcinoma and carcinoma surrounding prostatic tissue. METHODS: RT-PCR and real-time PCR was used to measure the expression of POU5F1P1 relative to the expression of HPRT1 in cell lines, prostatic carcinoma and carcinoma surrounding prostatic tissue. The structure of the POU5F1P1 mRNA and the promoter sequence were elucidated by 5' RACE experiments. The POU5F1P1 protein was shown with immunohistochemistry on prostate tissue. RESULTS: POU5F1P1 was found to be the only member of the POU5F1 family to be expressed in prostate with over-expression in prostatic carcinoma compared to surrounding prostatic tissue probably because of an increased density of expressing cells. The POU5F1P1 expression is driven by a variety of promoter structures scattered over a genomic region of 860 kB. CONCLUSIONS: The over expression of POU5F1P1 in prostatic carcinoma in addition to its genomic location and the putative function of its gene product render POU5F1P1 a good candidate to harbour functional genetic variants which modulate prostatic cancer susceptibility. PMID- 20017165 TI - The developmental expression profile of PAX2 in the murine prostate. AB - BACKGROUND: Nine transcription factors comprise the PAX gene family that regulate organogenesis. The urogenital system of PAX2 null male mice fails to develop properly. PAX2 is overexpressed in PC3 cells. Therefore, PAX2 is implicated in both prostate organogenesis and cancer. However, the expression pattern/profile of PAX2 in the prostate is unknown. METHODS: PAX2/5/8 expression was surveyed in E16.5 male urogenital sinus (UGS) by RT-PCR. Prostate samples from 10 developmental stages in C3H male mice were used in quantitative reverse transcript PCR (Q-PCR) and Western blotting (WB). RT-PCR and WB measured PAX2 expression in prostatic lobes or UGS layers, to identify local-regional expression patterns. Cytoplasmic versus nuclear expression was examined by WB. A castration series in adult C3H male mice and R1881 treatment in serum-free LNCaP cells examined androgen control of PAX2. RESULTS: PAX2 mRNA levels are higher in early developmental stages as compared to postpubertal prostates. RT-PCR and/or WB indicated a dorsal epithelial-nuclear localization of PAX2. PAX2 mRNA and protein increase postcastration. R1881 decreases expression of PAX2 mRNA in LNCaP cells as compared to controls. CONCLUSIONS: The expression profile of PAX2 indicates that it may regulate early, androgen-independent stages of murine prostate development, particularly for dorsally derived prostate glands. PAX2 expression appears to be associated with a dorsally localized epithelial cell population that is castration insensitive and retains proliferative and differentiative potential. Such a population of cells may represent a subset of stem-like cells having some characteristics in common with castrate-resistant prostate cancer cells. PMID- 20017166 TI - Controversies in the treatment of high-risk prostate cancer--what is the optimal combination of hormonal therapy and radiotherapy: a review of literature. AB - BACKGROUND: In high-risk prostate carcinoma, there is controversy whether these patients should be treated with escalated-dose (> or =74 Gy) or conventional-dose radiotherapy (<74 Gy) combined with hormonal therapy. Furthermore, the issue of the optimal duration and timing of hormonal therapy are not well crystallized. PATIENTS AND METHODS: A search for evidence from randomized- and large non randomized studies in order to address these issues, was therefore initiated. For this purpose, MedLine, EMbase, and PubMed and the data base of the Dutch randomized dose-escalation trial, were consulted. RESULTS AND CONCLUSIONS: From this search it was concluded that the benefit of hormonal therapy in combination with conventional-dose radiotherapy (<74 Gy) in high-risk prostate cancer is evident (Level 2 evidence); Levels 2 and 3 evidence were provided by several studies supporting the use of escalated-dose radiotherapy in high-risk prostate cancer. For the combination of hormonal therapy with escalated-dose radiotherapy in these patients, there is Level 2 evidence for moderately escalated dose (74 Gy) and high escalated dose (> or =78 Gy). The optimal duration and timing of hormonal therapy are not well defined. More randomized-controlled trials and meta analyses are therefore needed to clearly determine the independent role of dose escalation in high-risk patients treated with hormonal therapy and the optimal duration and timing of hormonal therapy. PMID- 20017167 TI - Retrospective analysis of prostate cancer recurrence potential with tissue metabolomic profiles. AB - BACKGROUND: In clinical care of prostate cancer patients, an improved method to assess the risk of recurrence after surgical treatment is urgently needed. We aim to retrospectively evaluate the ability of ex vivo tissue magnetic-resonance spectroscopy-based metabolomic profiles to estimate the risk of recurrence. METHODS: PCa recurrence is defined biochemically as the detection of serum PSA after radical prostatectomy. Sixteen consecutive PCa-recurrent cases, those with an initial PSA increase of 0.69 +/- 0.26 ng/ml monitored 47.7 +/- 2.6 months after prostatectomy were paired by age and Gleason score with cases without recurrence of the same pathological and clinical stages (n = 16/each). We analyzed ex vivo intact-tissue spectroscopy results from these 48 individuals at the time of prostatectomy at 14T. From these spectra, we identified the 27 most common and intense spectral metabolic regions for statistical analyses. RESULTS: Principal component analysis (PCA) on these spectral regions from cases of clinical-stage-matched groups with and without recurrence identified four pathology-related principal components. Canonical analysis of these four and the first nine principal components for cases in the two groups defined metabolomic profiles as the canonical score that can differentiate the two groups with statistical significance. By applying the coefficients from PCA and canonical analysis to the pathological-stage-matched groups, recurrence was predicted with an accuracy of 78%. CONCLUSIONS: Results indicate the potential of tissue metabolomic profiles measured with ex vivo spectroscopy to identify PCa aggressiveness in terms of cancer recurrence. With further study, this may greatly contribute to the future design of clinical strategy for personalized treatment of PCa patients. PMID- 20017168 TI - On the chirality of self-assembled DNA octahedra. PMID- 20017170 TI - Highly enantioselective hydrophosphonylation of aldehydes: base-enhanced aluminum salalen catalysis. PMID- 20017169 TI - Determinants of ligand affinity and heme reactivity in H-NOX domains. PMID- 20017171 TI - Platinum sintering on H-ZSM-5 followed by chemometrics of CO adsorption and 2D pressure-jump IR spectroscopy of adsorbed species. PMID- 20017173 TI - The oxyallyl diradical: observation of the singlet and triplet state by negative ion photoelectron spectroscopy. PMID- 20017172 TI - Direct detection of oxygen ligation to the Mn(4)Ca cluster of photosystem II by X ray emission spectroscopy. PMID- 20017174 TI - Multiresponsive reversible gels based on charge-driven assembly. PMID- 20017175 TI - DHTP ligands for the highly ortho-selective, palladium-catalyzed cross-coupling of dihaloarenes with grignard reagents: a conformational approach for catalyst improvement. PMID- 20017176 TI - Time-resolved in situ diffraction study of the solvothermal crystallization of some prototypical metal-organic frameworks. PMID- 20017177 TI - Effects of partial confinement on the specificity of monomolecular alkane reactions for acid sites in side pockets of mordenite. PMID- 20017178 TI - Ion- and liquid-assisted grinding: improved mechanochemical synthesis of metal organic frameworks reveals salt inclusion and anion templating. PMID- 20017179 TI - Peptoid-Peptide hybrid backbone architectures. AB - Peptidomimetic oligomers and foldamers have received considerable attention for over a decade, with beta-peptides and the so-called peptoids (N-alkylglycine oligomers) representing prominent examples of such architectures. Lately, hybrid or mixed backbones consisting of both alpha- and beta-amino acids (alpha/beta peptides) have been investigated in some detail as well. The present Minireview is a survey of the literature concerning hybrid structures of alpha-amino acids and peptoids, including beta-peptoids (N-alkyl-beta-alanine oligomers), and is intended to give an overview of this area of research within the field of peptidomimetic science. PMID- 20017180 TI - Structural water drives self-assembly of organic rosette nanotubes and holds host atoms in the channel. AB - We reveal how water solvent determines the self-assembly pathway and stability of organic rosette nanotubes (RNTs) and show their possible functions, using three dimensional molecular theory of solvation (a.k.a. 3D-RISM). Structural water molecules penetrate the pockets on the RNT outer surface, form a wetting monolayer in the RNT channel and bridge RNT rosettes. We predict that the inner water shell might stabilize rare gas atoms inside the RNT channel, and envision molecular devices with RNT channels transporting water or holding guest molecules for targeted delivery. PMID- 20017181 TI - Chemical thermodynamics of ultrasound speed in solutions and liquid mixtures. AB - A comprehensive formalism is developed to treat thermodynamically speed of ultrasound data for solutions and liquid mixtures. For solutions, the apparent speed of ultrasound of a solute is introduced and proposed to take the place of empirically defined quantities. The partial speed of ultrasound of a solute is defined and related to the partial molar volume and partial molar isentropic compression. For liquid mixtures, the concept of speed of sound before mixing pure liquids is presented and used to define the change in speed of ultrasound upon ideal mixing, which is predicted to be generally a negative quantity. A new thermodynamic equation is derived linking the values for excess speed of ultrasound, excess molar volume and excess molar isentropic compression of a mixture, and its applications are discussed. Ideal and excess apparent speeds of ultrasound, as well as ideal and excess partial speeds of ultrasound, are defined for substances making up a liquid mixture. Accurate speeds of ultrasound in 31 mixtures of water with the amphiphile 2-(ethylamino)ethanol at 293.15 K are reported. These data are used to demonstrate the ability of the apparent speed of ultrasound to describe the impact of solutes on sonic properties of solutions and the advantages of analysing thermodynamic properties of binary liquid mixtures in terms of the dependence on composition of Balankina's ratios between excess and ideal values. It is concluded that the new thermodynamic functions defined for speeds of ultrasound in solutions and liquid mixtures give, at the least, equivalent information on molecular aspects to the usual functions related to the isentropic compressibility, without needing density data for this purpose. PMID- 20017182 TI - Electrochemical machining of gold microstructures in LiCl/dimethyl sulfoxide. AB - LiCl/dimethyl sulfoxide (DMSO) electrolytes were applied for the electrochemical micromachining of Au. Upon the application of short potential pulses in the nanosecond range to a small carbon-fiber electrode, three-dimensional microstructures with high aspect ratios were fabricated. We achieved machining resolutions down to about 100 nm. In order to find appropriate machining parameters, that is, tool and workpiece rest potentials, the electrochemical behavior of Au in LiCl/DMSO solutions with and without addition of water was studied by cyclic voltammetry. In waterless electrolyte Au dissolves predominantly as Au(I), whereas upon the addition of water the formation of Au(III) becomes increasingly important. Because of the low conductivity of LiCl/DMSO compared with aqueous electrolytes, high machining precision is obtained with moderately short pulses. Furthermore, the redeposition of dissolved Au can be effectively avoided, since Au dissolution in LiCl/DMSO is highly irreversible. Both observations render LiCl/DMSO an appropriate electrolyte for the routine electrochemical micromachining of Au. PMID- 20017183 TI - Fibrinogen nanofibril growth and self-assembly on Au (1,1,1) surface in the absence of thrombin. PMID- 20017184 TI - Mechanism of the aerobic oxidation of alpha-pinene. AB - A combined experimental and theoretical approach is used to study the thermal autoxidation of alpha-pinene. Four different types of peroxyl radicals are generated; the verbenyl peroxyl radical being the most abundant one. The peroxyl radicals propagate a long radical chain, implying that chain termination does not play an important role in the production of the products. Two distinct types of propagation steps are active in parallel: the abstraction of allylic H atoms and the addition to the unsaturated C=C bond. The efficiency for both pathways appears to depend on the structure of the peroxyl radical. The latter step yields the corresponding epoxide product, as well as alkoxyl radicals. Under the investigated reaction conditions the alkoxyl radicals seem to produce both the alcohol and ketone products, the ketone presumably being formed upon the abstraction of the weakly bonded alphaH atom by O2. This mechanism explains the predominantly primary nature of all quantified products. At higher conversion, co oxidation of the hydroperoxide products constitutes an additional, albeit small, source of alcohol and ketone products. PMID- 20017185 TI - Endothelial IQGAP1 regulates efficient lymphocyte transendothelial migration. AB - Leukocyte movement from the blood to the tissues is a fundamental process in acute and chronic inflammatory diseases. While the role of endothelial cells (EC) to recruit leukocytes to sites of inflammation is well known, the mechanisms that control remodeling of EC shape and adhesive contacts during leukocyte transendothelial migration (TEM) are not completely understood. We studied the role of IQGAP1, an adaptor protein that binds to filamentous-actin and microtubules (MT) at interendothelial junctions, during lymphocyte TEM. EC IQGAP1 knockdown decreases MT tethered to the adherens junction, and decreases lymphocyte TEM to approximately 70% (p<0.05) versus control. Similarly, loss of adherens junction-associated MT induced by brief nocodazole (ND) treatment decreases lymphocyte TEM to approximately 65% of control (p<0.01). Confocal microscopy imaging indicates that EC IQGAP1 knockdown and MT depolymerization both result in lymphocyte accumulation above the vascular endothelial cadherin (VE-cadherin) junctions and reduces the fraction of adherent lymphocytes that complete diapedesis across interendothelial cell junctions. However, we observe no change in VE-cadherin gap formation underlying adherent lymphocytes among control, IQGAP1 knockdown, or MT depolymerised EC monolayers. These data indicate that IQGAP1 contributes to MT stability at endothelial junctions. Further, IQGAP1 is involved in junction remodeling required for efficient lymphocyte diapedesis, independent of VE-cadherin gap formation. PMID- 20017186 TI - Case-control study of pleural mesothelioma in workers with social security in Mexico. AB - BACKGROUND: Environmental and occupational exposure to asbestos in Mexico in the past has been a cause of deaths and health damages. Its magnitude is unknown to date. Our objective was to identify the proportion of cases of malignant pleural mesothelioma (MPM) that can be attributed to and occupational exposure to asbestos. METHODS: We carried out a case-control study of MPM in 472 workers insured by the Mexican Institute of Social Security, all Valley of Mexico residents, with 119 incident cases and 353 controls. Cases were histologically confirmed. Participants were questioned concerning their occupational history and sociodemographic data. Assignment to one of the four exposures was performed qualitatively by an expert hygienist. Odds ratios (ORs) and attributable risks (ARs) were calculated using a non-conditional logistic regression model. RESULTS: A total of 80.6% of cases and 31.5% of controls had occupational exposure to asbestos. ORs were adjusted for age and gender and by exposure category, and exhibited an increase with probability of exposure as follows: 3.7(95% CI 1.3 10.4) for the likely category and 14.3(95% CI 8-26) for the certain category; AR in the group occupationally exposed to asbestos was 83.2%, and the population AR was 44%. CONCLUSIONS: Our results show that the relationship between industrial uses of all forms of asbestos is generating an increase in mesothelioma-related diseases and deaths among Mexican workers. As a public health policy, Mexico should prohibit the use of asbestos in all production processes with the aim of controlling the epidemic and preventing the occurrence of new cases of MPM. PMID- 20017187 TI - Follow-up of neck and shoulder pain among sewing machine operators: The Los Angeles garment study. AB - BACKGROUND: The aim of the present study is to explore factors affecting or modifying self-reported neck/shoulder pain in sewing machine operators. METHODS: We investigated self-report neck/shoulder pain in 247 workers who participated in a 4-month prospective intervention study for musculoskeletal disorders. All participants were immigrants. We examine the influence of individual and work related factors on changes in neck/shoulder pain during follow-up employing linear mixed models with time-spline functions. RESULTS: We observed a dramatic decline (72%) in self-reported pain intensity in the first month of follow-up, followed by a small increase from the first to fourth month (4% per month). Workers who perceived and reported their physical workload as high or worked overtime experienced less overall pain reduction. Higher baseline pain intensity, being of Hispanic ethnicity (vs. Asian), and taking cumulative daily rest time during work of 35 min or more allowing for muscles to rest were associated with a larger pain reduction in the first month, but not thereafter. CONCLUSION: Our findings indicate that some work-related factors may be of clinical relevance for reducing neck/shoulder pain. Having lower physical workloads and less overtime work should be considered when treating patients or planning workplace interventions for managing work-related musculoskeletal disorders in this underserved immigrant population. PMID- 20017189 TI - Hearing problems among a cohort of nationally certified EMS professionals. AB - PURPOSE: The objectives of this study were to estimate the prevalence of hearing problems among a national cohort of emergency medical service (EMS) professionals, determine factors associated with hearing problems, and estimate the percentage of EMS professionals who utilize hearing protection. METHODS: Utilizing results from a questionnaire, individuals who reported hearing problems were compared to individuals who had not. Multivariable logistic regression was performed to identify variables associated with hearing problems. Finally, items regarding use of hearing protection were assessed to estimate the percentage of usage among EMS professionals. RESULTS: In total, 1,058 (57%) participants responded to the questionnaire. Of those, 1,024 (97%) who completed the hearing problems question were utilized for analysis. There were 153 (14.9%) cases of self-reported hearing problems. The final logistic regression model included lifetime occupational noise exposure 0.99 (95% CI = 0.9997-1.0002), report of previous back problems (odds ratio (OR) = 2.74, 95% CI = 1.8340-4.1042), large community size (OR = 1.67, 95% CI = 1.1211-2.4843), and minority status (OR = 0.61, 95% CI = 0.3719-0.9867). Finally, 213 (20.8%) individuals reported utilizing some form of hearing protection at their main EMS job. CONCLUSION: The results from this analysis are the first national estimates of the prevalence of self reported hearing problems among EMS professionals. This study was also the first to estimate the percentage of EMS professionals who self reported the utilization of hearing protection. PMID- 20017188 TI - Modified vaccinia Ankara-expressing Ag85A, a novel tuberculosis vaccine, is safe in adolescents and children, and induces polyfunctional CD4+ T cells. AB - Modified vaccinia Ankara-expressing Ag85A (MVA85A) is a new tuberculosis (TB) vaccine aimed at enhancing immunity induced by BCG. We investigated the safety and immunogenicity of MVA85A in healthy adolescents and children from a TB endemic region, who received BCG at birth. Twelve adolescents and 24 children were vaccinated and followed up for 12 or 6 months, respectively. Adverse events were documented and vaccine-induced immune responses assessed by IFN-gamma ELISpot and intracellular cytokine staining. The vaccine was well tolerated and there were no vaccine-related serious adverse events. MVA85A induced potent and durable T-cell responses. Multiple CD4+ T-cell subsets, based on expression of IFN-gamma, TNF-alpha, IL-2, IL-17 and GM-CSF, were induced. Polyfunctional CD4+ T cells co-expressing IFN-gamma, TNF-alpha and IL-2 dominated the response in both age groups. A novel CD4+ cell subset co-expressing these three Th1 cytokines and IL-17 was induced in adolescents, while a novel CD4+ T-cell subset co-expressing Th1 cytokines and GM-CSF was induced in children. Ag-specific CD8+ T cells were not detected. We conclude that in adolescents and children MVA85A safely induces the type of immunity thought to be important in protection against TB. This includes induction of novel Th1-cell populations that have not been previously described in humans. PMID- 20017190 TI - Tapasin edits peptides on MHC class I molecules by accelerating peptide exchange. AB - The endoplasmic reticulum (ER) protein tapasin is essential for the loading of high-affinity peptides onto MHC class I molecules. It mediates peptide editing, i.e. the binding of peptides of successively higher affinity until class I molecules pass ER quality control and exit to the cell surface. The molecular mechanism of action of tapasin is unknown. We describe here the reconstitution of tapasin-mediated peptide editing on class I molecules in the lumen of microsomal membranes. We find that in a competitive situation between high- and low-affinity peptides, tapasin mediates the binding of the high-affinity peptide to class I by accelerating the dissociation of the peptide from an unstable intermediate of the binding reaction. PMID- 20017192 TI - IL-4 induces protection of vascular endothelial cells against killing by complement and melittin through lipid biosynthesis. AB - We have shown previously that cytokines IL-4 and IL-13 induce protection in porcine vascular endothelial cells (EC) against killing by the membrane attack complex (MAC) of human complement. This protection is intrinsic, not due to changes in complement regulatory proteins, and requires activation of Akt and sterol receptor element binding protein-1 (SREBP-1), which regulates fatty acid and phospholipid synthesis. Here we report that, compared to EC incubated in medium, IL-4-treated EC had a profound reduction in complement-mediated ATP loss and in killing assessed by vital dye uptake, but only a slight reduction in permeability disruption measured by calcein release. While controls exposed to complement lost mitochondrial membrane potential and subsequently died, protected EC maintained mitochondrial morphology and membrane potential, and remained alive. SREBP-1 and fatty acid synthase activation were required for protection and fatty acid and phospholipid synthesis, including cardiolipin, were increased after IL-4 stimulation, without increase in cholesterol content or cell proliferation. IL-4 also induced protection of EC from killing by the channel forming protein melittin, similar to protection observed for the MAC. We conclude that IL-4 induced activation of Akt/SREBP-1/lipid biosynthesis in EC, resulting in protection against MAC and melittin, in association with mitochondrial protection. PMID- 20017191 TI - C5a receptor-deficient dendritic cells promote induction of Treg and Th17 cells. AB - C5a is a proinflammatory mediator that has recently been shown to regulate adaptive immune responses. Here we demonstrate that C5a receptor (C5aR) signaling in DC affects the development of Treg and Th17 cells. Genetic ablation or pharmacological targeting of the C5aR in spleen-derived DC results in increased production of TGF-beta leading to de novo differentiation of Foxp3(+) Treg within 12 h after co-incubation with CD4(+) T cells from DO11.10/RAG2(-/-) mice. Stimulation of C5aR(-/-) DC with OVA and TLR2 ligand Pam(3)CSK(4) increased TGF beta production and induced high levels of IL-6 and IL-23 but only minor amounts of IL-12 leading to differentiation of Th cells producing IL-17A and IL-21. Th17 differentiation was also found in vivo after adoptive transfer of CD4(+) Th cell into C5aR(-/-) mice immunized with OVA and Pam(3)CSK(4). The altered cytokine production of C5aR(-/-) DC was associated with low steady state MHC class II expression and an impaired ability to upregulate CD86 and CD40 in response to TLR2. Our data suggest critical roles for C5aR in Treg and Th17-cell differentiation through regulation of DC function. PMID- 20017193 TI - Thioredoxin suppresses airway inflammation independently of systemic Th1/Th2 immune modulation. AB - Oxidative stress plays an important role in the pathogenesis of asthma via the upregulation of local inflammatory mediators and/or promoting Th2-skewing during Ag sensitization. Thioredoxin (TRX), a 12 kDa redox-active protein with antioxidative property, has been recently shown to play a protective role in various inflammatory diseases. Using a mouse model of asthma, we show here that IL-13 and eotaxin production are decreased in TRX-Tg mice leading to reduced eosinophils recruitment and mucus metaplasia. The reduction in airway inflammation occurs without the attenuation of systemic Th2 immunity in that comparable levels of Th2-type cytokines and Ig were detected in LN and serum, respectively, from TRX-Tg and WT mice. Likewise, CD4(+) T cells from both strains of mice developed similar Th1 and Th2 responses in vitro. Asthmatic lungs of TRX Tg and WT mice contained similar amounts of GATA-3(+) and Foxp3(+) T cells. Finally, production of MIF, an upstream modulator of airway inflammation, was significantly reduced in the lungs of TRX-Tg mice. Our data suggest that TRX suppresses airway inflammation by inhibiting MIF production thereby limiting the downstream recruitment of eosinophils to the lung independently of modulating systemic Th1/Th2 immunity. PMID- 20017194 TI - GITR contributes to the systemic adjuvanticity of the Escherichia coli heat labile enterotoxin. AB - The Escherichia coli heat-labile enterotoxin (LT) possesses a powerful mucosal and systemic adjuvant effect. However, little is known about the cellular and molecular basis of the immunostimulatory activity of LT at the mucosal level, and even less information is available on the mechanisms underlying its systemic adjuvant activity. In this study, we show that distinct mechanisms are responsible for the parenteral and mucosal adjuvanticity of LT. Indeed, the systemic administration of LT upregulates the expression of glucocorticoid induced TNFR-related protein (GITR), but not other activation markers, in naive T cells. Using WT and GITR-deficient mice and LT and its enzymatically inactive mutant LTK63 as adjuvants, we show that the induction of GITR expression in T cells accounts for the systemic immunostimulatory capacity of LT, which requires an intact enzymatic activity. In contrast, the mucosal administration of LT does not induce GITR expression on Peyer's patche T cells and accordingly no differences are observed in the mucosal adjuvanticity of LT between WT and GITR deficient mice. Altogether, our results demonstrate the distinct effect of LT after parenteral administration when compared with the mucosal delivery, and describe a new mechanism of LT adjuvanticity related to its ability to induce the expression of GITR in CD4(+) T cells. PMID- 20017195 TI - Energy addiction and lymphocyte differentiation: a new role for the liver kinase B1 kinase. AB - Lymphocyte development is a process in which proliferation is coupled to differentiation. In order to undergo efficient proliferation, lymphocytes must coordinate the entry into cell cycle with increased metabolism. The signaling pathways, like those downstream of antigen and cytokine receptors, and specific regulators that directly control cell metabolism are only beginning to be defined. A study in this issue of the European Journal of Immunology, demonstrates that the liver kinase B1 (LKB1) is a regulator of cellular metabolism. Deficiency in LKB1 results in a block in T-cell differentiation, increased apoptosis, as well as impaired survival of mature T cells. This study highlights the importance of LKB1 in T-cell development and function, but as discussed in this Commentary, a number of intriguing questions concerning the regulation and additional functions of LKB1 remain. PMID- 20017196 TI - Mycobacterium tuberculosis Rv1419 encodes a secreted 13 kDa lectin with immunological reactivity during human tuberculosis. AB - In this study, we have identified a secreted 13 kDa lectin from Mtb (Mtb, Mycobacterium tuberculosis; sMTL-13) by homology search of a non-redundant lectin database. Bioinformatic analysis revealed that sMTL-13 belongs to the ricin-type beta-trefoil family of proteins containing a Sec-type signal peptide present in Mtb complex species, but not in non-tuberculous mycobacteria. Following heterologous expression of sMTL-13 and generation of an mAb (clone 276.B7/IgG1kappa), we confirmed that this lectin is present in culture filtrate proteins from Mtb H37Rv, but not in non-tuberculous mycobacteria-derived culture filtrate proteins. In addition, sMTL-13 leads to an increased IFN-gamma production by PBMC from active tuberculosis (ATB) patients. Furthermore, sera from ATB patients displayed high titers of IgG Ab against sMTL-13, a response found to be decreased following successful anti-tuberculosis therapy. Together, our findings reveal a secreted 13 kDa ricin-like lectin from Mtb, which is immunologically recognized during ATB and could serve as a biomarker of disease treatment. PMID- 20017197 TI - Intrathecal immune responses to EBV in early MS. AB - EBV has been consistently associated with MS, but its signature in the CNS has rarely been examined. In this study, we assessed EBV-specific humoral and cellular immune responses in the cerebrospinal fluid (CSF) of patients with early MS, other inflammatory neurological diseases (OIND) and non-inflammatory neurological diseases (NIND). The neurotropic herpesvirus CMV served as a control. Virus-specific humoral immune responses were assessed in 123 consecutive patients and the intrathecal recruitment of virus-specific antibodies was expressed as antibody indexes. Cellular immune responses tested in the blood of 55/123 patients were positive in 46/55. The CD8(+) CTL responses of these 46 patients were assessed in the blood and CSF using a CFSE-based CTL assay. We found that viral capsid antigen and EBV-encoded nuclear antigen-1, but not CMV IgG antibody indexes, were increased in early MS as compared with OIND and NIND patients. There was also intrathecal enrichment in EBV-, but not CMV-specific, CD8(+) CTL in early MS patients. By contrast, OIND and NIND patients did not recruit EBV- nor CMV-specific CD8(+) CTL in the CSF. Our data, showing a high EBV , but not CMV-specific intrathecal immune response, strengthen the association between EBV and MS, in particular at the onset of the disease. PMID- 20017198 TI - Effects of self-reported health conditions and pesticide exposures on probability of follow-up in a prospective cohort study. AB - BACKGROUND: We investigated the potential for selection bias due to non participation in the follow-up of a large prospective cohort study. METHODS: Licensed pesticide applicators (52,395 private; 4,916 commercial) in the Agricultural Health Study provided demographic, health, and pesticide exposure information at enrollment (1993-1997) and in a 5-year follow-up telephone interview. Factors associated with non-participation in the follow-up were identified using multiple logistic regression. Potential for selection bias was evaluated by comparing exposure-disease associations between the entire cohort and the follow-up subset. RESULTS: Sixty-six percent of private and 60% of commercial applicators completed the follow-up interview. Private and commercial applicators who did not complete the follow-up reported at enrollment younger age, less education, lower body mass index, poorer health behaviors but fewer health conditions, and lower pesticide use. Estimates of exposure-disease associations calculated with and without non-participants did not indicate strong selection bias. CONCLUSIONS: Differences between non-participants and participants in the follow-up interview were generally small, and we did not find significant evidence of selection bias. However, the extent of bias may depend on the specific exposure and outcome under study. PMID- 20017199 TI - Functions and therapeutic value of focal adhesion kinase signaling during hepatocellular carcinoma development and progression. PMID- 20017200 TI - Huh-7: a human "hemochromatotic" cell line. AB - Hereditary hemochromatosis (HC) is commonly associated with homozygosity for the cysteine-to-tyrosine substitution at position 282 (C282Y) of the HFE protein. This mutation prevents HFE from binding beta(2)-microglobulin (beta(2)M) and reaching the cell surface. We have discovered that a widely used hepatoma cell line, Huh-7, carries a HFE mutation similar to that associated with human HC. By HFE gene sequencing of Huh-7 genomic DNA, we found a TAC nucleotide deletion (c. 691_693del) responsible for loss of a tyrosine at position 231 (p. Y231del) of the HFE protein. This mutation affects a conserved hydrophobic region in a loop connecting two beta strands that make up the alpha3 domain of HFE, not far from the 282 site. HFE was detected by western blot in HepG2 but not in Huh-7 cell membrane fractions. In WRL-68 cells expressing wild-type HFE, the HFE protein was largely found at the plasma membrane where it colocalizes with beta(2)M. On the contrary, the HFE-Y231del mutant, similarly to an exogenously expressed HFE-C282Y mutant, failed to reach the plasma membrane and did not colocalize with membrane expressed beta(2)M. C282Y mutant HFE in HC is associated with inadequate hepcidin expression. We found that Huh-7 cells display lower hepcidin messenger RNA levels as compared to HepG2 cells, which carry a wild-type HFE. Interestingly, hepcidin messenger RNA levels increased significantly in Huh-7 cells stably expressing exogenous wild-type HFE at the plasma membrane. CONCLUSION: Huh-7 cells may represent a novel and valuable tool to investigate the role of altered HFE traffic in iron metabolism and pathogenesis of human HFE HC. PMID- 20017201 TI - Free deep circumflex iliac artery vascularised bone flap for reconstruction of the distal radius: planning with CT angiography. AB - Distal radius fractures in the younger population are often comminuted and intra articular, which can increase the complexity of their management. In addition, these patients tend to place high demands on their wrists, and the prevention of functional arthritis necessitates excellent anatomical reduction. Complicated cases such as these are often limited in their management options. We present a complex case of distal radius fracture and bone loss in which initial therapy with nonvascularized bone graft failed, and osteomyelitis was a further complicating factor. With the aid of preoperative planning with computed tomographic angiography (CTA), a deep circumflex iliac artery (DCIA) bone flap was able to be assessed as a reconstructive option. The use of preoperative CTA, the first description of such imaging in this role, was able to delineate the bone to be harvested, confirm its vascular supply, and plan flap harvest. The use of a vascularized bone flap in this setting was thus undertaken and was able to provide an autologous anatomical support for the wrist while reducing the risk of recurrent infection and still preserving internal fixation. This unique application of the free DCIA bone flap was potentiated by CTA, achieving complete healing and good functional outcomes. PMID- 20017202 TI - Failed digital nerve reconstruction by foreign body reaction to Neurolac nerve conduit. PMID- 20017203 TI - alphaPS1betaPS integrin receptors regulate the differential distribution of Sog fragments in polarized epithelia. AB - Bone morphogenetic proteins (BMPs) have important functions during epithelial development. In Drosophila, extracellular Short gastrulation (Sog) limits the action of the BMP family member Decapentaplegic (Dpp). We have shown that Integrin receptors regulate Sog activity and distribution during pupal wing development to direct placement of wing veins. Here, we show that Integrins perform a similar function in the follicular epithelium, impacting Dpp function during oogenesis and embryonic development. As reported for the wing, this effect is specific to mew, which codes for alphaPS1 integrin. Sog is subject to cleavage by metalloproteases, generating fragments with different properties. We also show that Integrins regulate the distribution of C- and N-terminal Sog fragments in both epithelia, suggesting they may regulate the quality of BMP outputs. Our data indicate that alphaPS1betaPS integrin receptors regulate the amount and type of Sog fragments available for diffusion in the extracellular space during oogenesis and pupal wing development. PMID- 20017206 TI - The role of entrepreneurial activities in academic pharmaceutical science research. AB - Academic pharmaceutical science research is expanding further and further from the University setting to encompass the for-profit private company setting. This parallels the National Institutes of Health momentum to include multiple funding opportunities for University and private company collaboration. It has been recognized that the nonprofit and for-profit combination research model can accelerate the commercialization of pharmaceutical products, and therefore more efficiently improve human health. Entrepreneurial activities require unique considerations in the University environment, but can be modeled after the commercialization expansion of the academic healthcare enterprise. Challenges and barriers exist to starting a company as an entrepreneurial faculty member, but the rewards to one's personal and professional lives are incomparable. PMID- 20017204 TI - Initial testing of topotecan by the pediatric preclinical testing program. AB - BACKGROUND: Topotecan is a small molecule DNA topoisomerase I poison, that has been successful in clinical trials against pediatric solid tumors and leukemias. Topotecan was evaluated against the Pediatric Preclinical Testing Program (PPTP) tumor panels as part of a validation process for these preclinical models. PROCEDURES: In vivo three measures of antitumor activity were used: (1) an objective response measure modeled after the clinical setting; (2) a treated to control (T/C) tumor volume measure; and (3) a time to event (fourfold increase in tumor volume for solid tumor models, or > or =25% human CD45+ cells in the peripheral blood for acute lymphoblastic leukemia, ALL models) measure based on the median event-free survival (EFS) of treated and control animals for each xenograft. RESULTS: Topotecan inhibited cell growth in vitro with IC(50) values between 0.71 and 489 nM. Topotecan significantly increased EFS in 32 of 37 (87%) solid tumor xenografts and in all 8 of the ALL xenografts. Seventy-five percent of solid tumors met EFS T/C activity criteria for intermediate (n = 17) or high activity (n = 7). Objective responses were noted in eight solid tumor xenografts (Wilms, rhabdomyosarcoma, Ewing sarcoma, neuroblastoma). Among the six neuroblastomas, three achieved a PR. For the ALL panel, two maintained CRs, three CRs, and two PRs were observed. CONCLUSIONS: Topotecan demonstrated broad activity in vitro and in vivo against both the solid tumor and ALL panels, with significant tumor growth delay generated in all the panels. These results further demonstrate the validity of the PPTP panel for preclinical testing of new drugs. PMID- 20017207 TI - Topography of the chorda tympani nerve and the tensor tympani muscle in carnivores provides a new synapomorphy for Herpestidae (Carnivora, Mammalia). AB - The topographical relationship of the chorda tympani nerve (chorda tympani) to the tensor tympani muscle in the middle ear of carnivores provides new phylogenetic information. The examination of histological serial sections of 16 carnivore species representing most families revealed two distinct character states concerning the course of the chorda tympani: a hypotensoric state with the nerve running below the insertion tendon of the tensor tympani muscle, and an epitensoric state with the nerve running above the tendon. The shift from the plesiomorphic hypotensoric chorda tympani to the apomorphic epitensoric condition occurred once in carnivore phylogeny: Only in the herpestid species under study does the chorda tympani cross above the tensor tympani muscle. Therefore, we introduce the epitensoric pattern as a new synapomorphy for herpestids. Within the herpestids we find the following structural distinctions: Herpestes javanicus and Galerella sanguinea have a chorda tympani running in a sulcus directly above the insertion of the tensor tympani muscle, whereas in the eusocial herpestid species Suricata suricatta and Mungos mungo the chorda tympani lies far above the insertion of the muscle. PMID- 20017208 TI - Patterned assembly and neurogenesis in the chick dorsal root ganglion. AB - The birth of small-diameter TrkA+ neurons that mediate pain and thermoreception begins approximately 24 hours after the cessation of neural crest cell migration from progenitors residing in the nascent dorsal root ganglion. Although multiple geographically distinct progenitor pools have been proposed, this study is the first to comprehensively characterize the derivation of small-diameter neurons. In the developing chick embryo we identify novel patterns in neural crest cell migration and colonization that sculpt the incipient ganglion into a postmitotic neuronal core encapsulated by a layer of proliferative progenitor cells. Furthermore, we show that this outer progenitor layer is composed of three spatially, temporally, and molecularly distinct progenitor zones, two of which give rise to distinct populations of TrkA+ neurons. PMID- 20017209 TI - Expression of the paralogous tyrosine hydroxylase encoding genes th1 and th2 reveals the full complement of dopaminergic and noradrenergic neurons in zebrafish larval and juvenile brain. AB - The development of dopaminergic and noradrenergic neurons has received much attention based on their modulatory effect on many behavioral circuits and their involvement in neurodegenerative diseases. The zebrafish (Danio rerio) has emerged as a new model organism with which to study development and function of catecholaminergic systems. Tyrosine hydroxylase is the entry enzyme into catecholamine biosynthesis and is frequently used as a marker for catecholaminergic neurons. A genome duplication at the base of teleost evolution resulted in two paralogous zebrafish tyrosine hydroxylase-encoding genes, th1 and th2, the expression of which has been described previously only for th1. Here we investigate the expression of th2 in the brain of embryonic and juvenile zebrafish. We optimized whole-mount in situ hybridization protocols to detect gene expression in the anatomical three-dimensional context of whole juvenile brains. To confirm whether th2-expressing cells may indeed use dopamine as a neurotransmitter, we also included expression of dopamine beta hydroxylase, dopa decarboxylase, and dopamine transporter in our analysis. Our data provide the first complete account of catecholaminergic neurons in the zebrafish embryonic and juvenile brain. We identified four major th2-expressing neuronal groups that likely use dopamine as transmitter in the zebrafish diencephalon, including neurons of the posterior preoptic nucleus, the paraventricular organ, and the nuclei of the lateral and posterior recesses in the caudal hypothalamus. th2 expression in the latter two groups resolves a previously reported discrepancy, in which strong dopamine but little tyrosine hydroxylase immunoreactivity had been detected in the caudal hypothalamus. Our data also confirm that there are no mesencephalic DA neurons in zebrafish. PMID- 20017210 TI - Genetic dissection of dopaminergic and noradrenergic contributions to catecholaminergic tracts in early larval zebrafish. AB - The catecholamines dopamine and noradrenaline provide some of the major neuromodulatory systems with far-ranging projections in the brain and spinal cord of vertebrates. However, development of these complex systems is only partially understood. Zebrafish provide an excellent model for genetic analysis of neuronal specification and axonal projections in vertebrates. Here, we analyze the ontogeny of the catecholaminergic projections in zebrafish embryos and larvae up to the fifth day of development and establish the basic scaffold of catecholaminergic connectivity. The earliest dopaminergic diencephalospinal projections do not navigate along the zebrafish primary neuron axonal scaffold but establish their own tracts at defined ventrolateral positions. By using genetic tools, we study quantitative and qualitative contributions of noradrenergic and defined dopaminergic groups to the catecholaminergic scaffold. Suppression of Tfap2a activity allows us to eliminate noradrenergic contributions, and depletion of Otp activity deletes mammalian A11-like Otp dependent ventral diencephalic dopaminergic groups. This analysis reveals a predominant contribution of Otp-dependent dopaminergic neurons to diencephalospinal as well as hypothalamic catecholaminergic tracts. In contrast, noradrenergic projections make only a minor contribution to hindbrain and spinal catecholaminergic tracts. Furthermore, we can demonstrate that, in zebrafish larvae, ascending catecholaminergic projections to the telencephalon are generated exclusively by Otp-dependent diencephalic dopaminergic neurons as well as by hindbrain noradrenergic groups. Our data reveal the Otp-dependent A11-type dopaminergic neurons as the by far most prominent dopaminergic system in larval zebrafish. These findings are consistent with a hypothesis that Otp-dependent dopaminergic neurons establish the major modulatory system for somatomotor and somatosensory circuits in larval fish. PMID- 20017211 TI - Distribution of leptin-sensitive cells in the postnatal and adult mouse brain. AB - Leptin plays a pivotal role in the regulation of energy homeostasis and neuroendocrine functions, and increasing evidence indicates that leptin acts on the brain to mediate many of these effects. Recent data have also suggested that leptin influences brain development during early postnatal life. Here we examined the distribution of cells that express mRNA encoding the long form of the leptin receptor (LepRb) in postnatal and adult mouse brains by using in situ hybridization. In both adults and neonates, LepRb mRNA was largely restricted to regions known to control energy balance. Labeled cells were found in the arcuate, ventromedial, and dorsomedial nuclei of the hypothalamus as well as in the lateral hypothalamic area. Heavily labeled cells were also found in the median preoptic and ventral premammillary nuclei, two hypothalamic nuclei that are known to control reproduction. Moreover, during postnatal and adult life, clearly labeled cells were found in extrahypothalamic autonomic control sites such as the nucleus of the tractus solitarius. Importantly, this receptor can induce intracellular signaling because peripheral injection of leptin caused STAT3 phosphorylation in most sites in which LepRb mRNA was expressed. LepRb mRNA was also transiently elevated in certain regions of the postnatal mouse brain, such as the cortex, hippocampus, and laterodorsal nucleus of the thalamus. Taken together, these observations are consistent with the proposed roles of leptin in feeding and neuroendocrine regulation. They also identify regions where LepRb mRNA is expressed during early postnatal life and suggest new roles for leptin in the nervous system during development. PMID- 20017212 TI - Fragmentation study of iridoid glycosides and phenylpropanoid glycosides in Radix Scrophulariae by rapid resolution liquid chromatography with diode-array detection and electrospray ionization time-of-flight mass spectrometry. AB - Rapid resolution liquid chromatography (RRLC) coupled with diode array detection (DAD) and electrospray ionization time-of-flight mass spectrometry (ESI-TOF MS) method was applied to the mass spectral study of a series of naturally occurring iridoid glycosides and phenylpropanoid glycosides in Radix Scrophulariae, which provides higher speed and increased sensitivity without loss of resolution. With dynamic adjustment as the key role of the fragmentor voltage and confirmed with authentic standards, valuable structural information regarding the nature of both the glycoside skeletons was thus obtained. Most compositions were found to possess organic acid moiety such as cinnamoyl, caffeoyl and ferulyol. Besides extensive fragmentation of the carbohydrate moiety, losses of the hydroxyl and glucose residue units showed in the spectra, permitting the exploration of the skeleton and the identity of substituents in the molecule. Ten major iridoid glycosides and 10 phenylpropanoid glycosides were identified or tentatively characterized based on their retention times, UV and TOF MS data. The major fragmentation pathways of PGs in Radix Scrophulariae obtained through the MS data was schemed systematically for the first time, which provides a reference for other PGs derivatives. PMID- 20017213 TI - Enantiomeric separation of 2-arylpropionic acid nonsteroidal anti-inflammatory drugs by HPLC with hydroxypropyl-beta-cyclodextrin as chiral mobile phase additive. AB - The enantio-separations of eight 2-arylpropionic acid nonsteroidal anti inflammatory drugs (2-APA NSAIDs) were established using reversed-phase high performance liquid chromatography with hydroxypropyl-beta-cyclodextrin (HP-beta CD) as chiral mobile phase additive for studying the stereoselective skin permeation of suprofen, ketoprofen, naproxen, indoprofen, fenoprofen, furbiprofen, ibuprofen and carprofen. The effects of the mobile phase composition, concentration of HP-beta-CD and column temperature on retention and enantioselective separation were investigated. With 2-APA NSAIDs as acidic analytes, the retention times and resolutions of the enantiomers were strongly related to the pH of the mobile phase. In addition, both the concentration of HP beta-CD and temperature had a great effect on retention time, but only a slight or almost no effect on resolutions of the analytes. Enantioseparations were achieved on a Shimpack CLC-ODS (150 x 4.6 mm i.d., 5 microm) column. The mobile phase was a mixture of methanol and phosphate buffer (pH 4.0-5.5, 20 mM) containing 25 mM HP-beta-CD. This method was flexible, simple and economically advantageous over the use of chiral stationary phase, and was successfully applied to the enantioselective determination of the racemic 2-APA NSAIDs in an enantioselective skin permeation study. PMID- 20017214 TI - Crystal structure of NADH:rubredoxin oxidoreductase from Clostridium acetobutylicum: a key component of the dioxygen scavenging system in obligatory anaerobes. PMID- 20017215 TI - De novo backbone scaffolds for protein design. AB - In recent years, there have been significant advances in the field of computational protein design including the successful computational design of enzymes based on backbone scaffolds from experimentally solved structures. It is likely that large-scale sampling of protein backbone conformations will become necessary as further progress is made on more complicated systems. Removing the constraint of having to use scaffolds based on known protein backbones is a potential method of solving the problem. With this application in mind, we describe a method to systematically construct a large number of de novo backbone structures from idealized topological forms in a top-down hierarchical approach. The structural properties of these novel backbone scaffolds were analyzed and compared with a set of high-resolution experimental structures from the protein data bank (PDB). It was found that the Ramachandran plot distribution and relative gamma- and beta-turn frequencies were similar to those found in the PDB. The de novo scaffolds were sequence designed with RosettaDesign, and the energy distributions and amino acid compositions were comparable with the results for redesigned experimentally solved backbones. PMID- 20017216 TI - DynaDock: A new molecular dynamics-based algorithm for protein-peptide docking including receptor flexibility. AB - Molecular docking programs play an important role in drug development and many well-established methods exist. However, there are two situations for which the performance of most approaches is still not satisfactory, namely inclusion of receptor flexibility and docking of large, flexible ligands like peptides. In this publication a new approach is presented for docking peptides into flexible receptors. For this purpose a two step procedure was developed: first, the protein-peptide conformational space is scanned and approximate ligand poses are identified and second, the identified ligand poses are refined by a new molecular dynamics-based method, optimized potential molecular dynamics (OPMD). The OPMD approach uses soft-core potentials for the protein-peptide interactions and applies a new optimization scheme to the soft-core potential. Comparison with refinement results obtained by conventional molecular dynamics and a soft-core scaling approach shows significant improvements in the sampling capability for the OPMD method. Thus, the number of starting poses needed for successful refinement is much lower than for the other methods. The algorithm was evaluated on 15 protein-peptide complexes with 2-16mer peptides. Docking poses with peptide RMSD values <2.10 A from the equilibrated experimental structures were obtained in all cases. For four systems docking into the unbound receptor structures was performed, leading to peptide RMSD values <2.12 A. Using a specifically fitted scoring function in 11 of 15 cases the best scoring poses featured a peptide RMSD < or = 2.10 A. PMID- 20017217 TI - Dicistronic regulation of fluorescent proteins in the budding yeast Saccharomyces cerevisiae. AB - Fluorescent proteins are convenient tools for measuring protein expression levels in the budding yeast Saccharomyces cerevisiae. Co-expression of proteins from distinct vectors has been seen by fluorescence microscopy; however, the expression of two fluorescent proteins on the same vector would allow for monitoring of linked events. We engineered constructs to allow dicistronic expression of red and green fluorescent proteins and found that expression levels of the proteins correlated with their order in the DNA sequence, with the protein encoded by the 5'-gene more highly expressed. To increase expression levels of the second gene, we tested four regulatory elements inserted between the two genes: the IRES sequences for the YAP1 and p150 genes, and the promoters for the TEF1 gene from both S. cerevisiae and Ashbya gossypii. We generated constructs encoding the truncated ADH1 promoter driving expression of the red protein, yeast enhanced Cherry, followed by a regulatory element driving expression of the green protein, yeast-enhanced GFP. Three of the four regulatory elements successfully enhanced expression of the second gene in our dicistronic construct. We have developed a method to express two genes simultaneously from one vector. Both genes are codon-optimized to produce high protein levels in yeast, and the protein products can be visualized by microscopy or flow cytometry. With this method of regulation, the two genes can be driven in a dicistronic manner, with one protein marking cells harbouring the vector and the other protein free to mark any event of interest. PMID- 20017218 TI - Paralympians--unknown heroes next door. PMID- 20017219 TI - Ensuring the integrity of the published record: Croatian Medical Journal endorses guidance on retractions in scientific journals by Committee on Publication Ethics (COPE). PMID- 20017220 TI - Retractions: guidance from the Committee on Publication Ethics (COPE). PMID- 20017221 TI - Alteration in TWIST expression: possible role in paclitaxel-induced apoptosis in human laryngeal carcinoma Hep-2 cell line. AB - AIM: To explore the relationship between alteration in the expression of TWIST, highly conserved transcription factor from the basic helix-loop-helix family, and apoptosis of Hep-2 cells induced by chemotherapeutic agent paclitaxel. METHODS: Morphological changes of Hep-2 cells were observed by acridine orange cytochemistry staining. Viability of Hep-2 cells treated with various concentrations of paclitaxel was examined by cell proliferation assay. Apoptosis was examined by flow cytometry. The mRNA and protein expression of TWIST in response to paclitaxel at 24 hours, 48 hours, and 72 hours was examined by reverse transcription-polymerase chain reaction (RT-PCR) and Western blotting, respectively. RESULTS: Typical morphological changes of apoptotic cells at 24 hours, 48 hours, or 72 hours after treatment wiyth paclitaxel (10x10(-9) mol/L) were observed. The cell survival rates significantly decreased in a concentration and time-dependent manner (P=0.001). Paclitaxel-induced apoptosis increased with culture time (22.6+/-5.3% after 24 hours, 38.7+/-7.9% after 48 hours, and 52.4+/ 14.3% after 72 hours; P=0.002). Both mRNA and protein expression of TWIST was markedly decreased at both mRNA levels and protein levels, at 24 hours, 48 hours, and 72 hours in the paclitaxel-induced apoptosis of Hep-2 cells (P<0.001). CONCLUSION: TWIST, which has a significantly decreased expression in response to paclitaxel in Hep-2 cells, may play a pivotal role in paclitaxel-induced apoptosis of Hep-2 cells. PMID- 20017222 TI - Predictors of increased left ventricular filling pressure in dialysis patients with preserved left ventricular ejection fraction. AB - AIM: To study the left and right ventricular function and to assess the predictors of increased left ventricular (LV) filling pressure in dialysis patients with preserved LV ejection fraction. METHODS: This study included 63 consecutive patients (age 57+/-14 years, 57% women) with end-stage renal failure. Echocardiography, including tissue Doppler measurements, was performed in all patients. Based on the median value of the ratio of transmitral early diastolic velocity to early myocardial velocity (E/E' ratio), patients were divided into 2 groups: the group with high filling pressure (E/E'>10.16) and the group with low filling pressure (E/E'< or =10.16). RESULTS: Compared with patients with low filling pressure, the group of patients with high filling pressure included a higher proportion of diabetic patients (41% vs 13%, P=0.022) and had greater LV mass index (211+/-77 vs 172+/-71 g/m3, P=0.04), lower LV lateral long axis amplitude (1.4+/-0.3 vs 1.6+/-0.3 cm, P=0.01), lower E wave (84+/-19 vs 64+/ 18cm/s, P<0.001), higher systolic myocardial velocity (S': 8.6+/-1. 5 vs 7.0+/ 1.3 cm/s, P<0.001), and lower diastolic myocardial velocities (E': 6.3+/-1.9 vs 9.5+/-2.9 cm/s, P<0.001; A': 8.4+/-1.9 vs 9.7+/-2.5 cm/s, P=0.018). Multivariate analysis identified LV systolic myocardial velocity - S' wave (adjusted odds ratio, 1.909; 95% confidence interval, 1.060-3.439; P=0.031) and age (1.053; 1.001-1.108; P=0.048) as the only independent predictors of high LV filling pressure in dialysis patients. CONCLUSIONS: In dialysis patients with preserved left ventricular ejection fraction, reduced systolic myocardial velocity and elderly age are independent predictors of increased left ventricular filling pressure. PMID- 20017223 TI - ABO blood groups and genetic risk factors for thrombosis in Croatian population. AB - AIM: To assess the association between ABO blood group genotypes and genetic risk factors for thrombosis (FV Leiden, prothrombin G20210A, and methylenetetrahydrofolate reductase C677T mutations) in the Croatian population and to determine whether genetic predisposition to thrombotic risk is higher in non-OO blood group genotypes than in OO blood group genotypes. METHODS: The study included 154 patients with thrombosis and 200 asymptomatic blood donors as a control group. Genotyping to 5 common alleles of ABO blood groups was performed by polymerase chain reaction with sequence specific primers (PCR-SSP). FV Leiden was determined by PCR-SSP, while prothrombin and methylenetetrahydrofolate reductase were determined by PCR and restriction fragment length polymorphism (PCR-RFLP). RESULTS: There was an association between non-OO blood group genotypes and the risk of thrombosis (odds ratio [OR] 2.08, 95% confidence interval [CI], 1.32-3.27). The strongest association with thrombotic risk was recorded for A1B/A2B blood group genotypes (OR, 2.73; 95% CI, 1.10-6.74), followed by BB/O1B/O2B (OR, 2.29; 95% CI, 1.25-4.21) and O1A1/O2A1 (OR, 1.95; 95% CI, 1.15-3.31). FV Leiden increased the risk of thrombosis 31-fold in the group of OO carriers and fourfold in the group of non-OO carriers. There was no significant difference in the risk of thrombosis between OO and non-OO blood groups associated with prothrombin mutation. Non-OO carriers positive for methylenetetrahydrofolate reductase had a 5.7 times greater risk of thrombosis than that recorded in OO carriers negative for methylenetetrahydrofolate reductase. CONCLUSION: Study results confirmed the association of non-OO blood group genotypes with an increased risk of thrombosis in Croatia. PMID- 20017224 TI - Higher serum uric acid on admission is associated with higher short-term mortality and poorer long-term survival after myocardial infarction: retrospective prognostic study. AB - AIM: To assess serum uric acid (SUA) levels determined on admission as a potential predictor of short-term mortality and long-term survival in acute myocardial infarction (AMI) patients. METHOD: Data for this retrospective prognostic study were drawn from the patient database of the Varazdin County General Hospital in Varazdin, Croatia. We included consecutive patients with verified AMI admitted within 48 hours since the symptom onset during the period between 1 January 1996 and 31 December 2001. Long-term survival/mortality data were collected through direct contacts with patients and search of the community death registries. Relative risks (RR) and hazard ratios (HR) by 10 micromol/L increase in SUA were determined using modified Poisson regression with robust error variance and proportional hazard regression, respectively. RESULTS: A total of 621 patients (age 27-90 years, 64.7% men, 77.5% AMI with ST elevation, SUA 63 993 micromol/L) were included. Higher SUA on admission was independently associated with higher in-hospital mortality (RR, 1.016; 95% confidence interval [CI], 1.001-1.031, P=0.043) and higher thirty-day mortality (RR, 1.016; 95% CI, 1.003-1.029, P=0.018). Considered covariates were demographics, pre-index event cardiovascular morbidity and treatment, on-admission serum creatinine, total cholesterol and triglycerides, AMI characteristics, and peak creatine phosphokinase. Higher SUA on admission was also independently associated with poorer long-term survival (ie, higher all-cause mortality) (HR, 1.105; 95% CI, 1.020-1.195, P=0.010). Considered covariates were demographics, laboratory variables on admission, AMI characteristics, peak creatine phosphokinase, acute complications, and treatment at discharge. CONCLUSION: Higher serum uric acid determined on admission is associated with higher in-hospital mortality and thirty-day mortality and poorer long-term survival after AMI. PMID- 20017225 TI - Knowledge and practices of obtaining informed consent for medical procedures among specialist physicians: questionnaire study in 6 Croatian hospitals. AB - AIM. To assess physicians' knowledge and practices for obtaining patients' informed consent to medical procedures. METHODS. An anonymous and voluntary survey of knowledge and practices for obtaining informed consent was conducted among 470 physicians (63% response rate) working in 6 hospitals: 93 specialists in anesthesiology, 166 in internal medicine, and 211 in surgery. RESULTS. Only 54% physicians were acquainted with the fact that the procedure for obtaining consent was regulated by the law. Internists and surgeons were better informed than anesthesiologists (P=0.024). More than a half of respondents (66%) were familiar with the fact that a law on patient rights was passed in Croatia; there were no differences among different specialties (P=0.638). Only 38% of the physicians were fully informed about the procedure of obtaining consent. Internists and surgeons provided detailed information to the patient in 33% of the cases and anesthesiologists in 16% of the cases (P<0.050). Internists reported spending more time on informing the patient than anesthesiologists and surgeons (P<0.001). There were no differences in knowledge and practices for obtaining informed consent between physicians working in university and those working in community hospitals (P> or =0.05 for all questions). CONCLUSION. Physicians in Croatia have no formal education on informed consent and implement the informed consent process in a rather formal manner, regardless of the type of hospital or medical specialty. Systemic approach at education and training at the national level is needed to improve the informed consent process. PMID- 20017228 TI - Potential problems in building social model of disability. PMID- 20017226 TI - Revolving-door patients in a public psychiatric hospital in Israel: cross sectional study. AB - AIM. To study social, demographic, clinical, and forensic profiles of frequently re-hospitalized (revolving-door) psychiatric patients. METHODS. The study included all patients (n=183) who were admitted to our hospital 3 or more times during a 2-year period from 1999 through 2000. We compared these patients to 2 control groups of patients who were admitted to our hospital in the same period. For comparison of forensic data, we compared them with all non revolving-door patients (n=1056) registered in the computerized hospital database and for comparison of medical and clinical data we compared them with a random sample of non revolving-door patients (n=98). The sample was sufficiently large to yield high statistical power (above 98%). We collected data on the legal status of the hospitalizations (voluntary or involuntary) and social, demographic, clinical, and forensic information from the forensic and medical records of revolving-door and non revolving-door patients. RESULTS. In the period 1999-2000, 183 revolving door patients accounted for 771 (37.8%, 4.2 admissions per patient) and 1056 non revolving-door patients accounted for 1264 (62.5%, 1.2 admissions per patient) of the 2035 admissions to our hospital. Involuntary hospitalizations accounted for 23.9% of revolving-door and 76.0% of non revolving-door admissions. Revolving door patients had significantly shorter mean interval between hospitalizations, showed less violence, and were usually discharged contrary to medical advice. We found no differences in sex, marital status, age, ethnicity, diagnoses, illegal drug and alcohol use, or previous suicide-attempts between the groups. CONCLUSIONS. Revolving-door patients are not necessarily hospitalized for longer time periods and do not have more involuntarily admissions. The main difference between revolving-door and non revolving-door patients is greater self-management of the hospitalization process by shortening the time between voluntary re admission and discharge against medical advice. PMID- 20017227 TI - Illness perception and cardiovascular risk factors in patients with type 2 diabetes: cross-sectional questionnaire study. AB - AIM. To investigate illness perception in patients with type 2 diabetes mellitus and its association with the degree of control over relevant cardiovascular risk factors. METHODS. A cross-sectional questionnaire-based study was performed from June 2007 to March 2008. A stratified random sample of 46 Croatian general practitioners was asked to select, using systematic sampling, the first 6 patients with type 2 diabetes mellitus aged > or =18 years who visited them for consultation during the study period. Data on 250 patients included patient illness perception assessment (Brief Illness Perception Questionnaire, IPQ), cardiovascular risk factors, and socio-demographic data. RESULTS. The patients' mean age was 63.0+/-10.9 years and mean duration of diabetes was 9.3+/-7.8 years. The patients' illness perception assessment on an 11-point (0 to 10) scale showed the highest median scores (interquartile range): 10 (8 to 10) for "timeline" and 8 (7 to 9) for "treatment control," followed by 7 (5 to 8) for "personal control," 7 (5 to 9) for "understanding," 5 (3 to 7) for "consequences," 6 (4 to 7) for "concern," and 5 (2 to 7) for "emotional response." The lowest score was 3 (1 to 5) for "identity." Multivariate logistic regression showed that the Brief IPQ item "concern" (P<0.001) was a significant predictor of body mass index; "personal control" (P<0.001) and "concern" (P=0.048) were significant predictors of fasting blood glucose; "treatment control" (P=0.009) was a significant predictor of total cholesterol; and "understanding" (P=0.010) was a significant predictor of blood pressure. CONCLUSION. As patients' beliefs seem to be associated with the degree of control over cardiovascular risk factors, they should be included in routine clinical assessments. PMID- 20017229 TI - Paradigm change from curative to predictive medicine: novel strategic trends in Europe. PMID- 20017230 TI - My Africa. Poverty, health, disease, and medical journalism. PMID- 20017231 TI - From Adamovec to Dubrovnik: an extended adventure, 1971-1992. PMID- 20017232 TI - Patient voice. Christmas 2009. PMID- 20017233 TI - Information from your family doctor. Electronic fetal monitoring. PMID- 20017234 TI - Northern exposure. Interview by Laura Gater. PMID- 20017235 TI - Information from your family doctor. Hematospermia: blood in the semen. PMID- 20017236 TI - Information from your family doctor. Hip impingement: what you should know. PMID- 20017237 TI - Information from your family doctor. Latex allergy. PMID- 20017238 TI - [The 112th annual meeting of the Japanese Ophthalmological Society. Yokohama, Japan. April 17-20, 2008]. PMID- 20017239 TI - [The 45th Annual Meeting of the Japanese Society of Nuclear Medicine. Sandai, Japan. November 4-6, 2007. Abstracts]. PMID- 20017240 TI - [The 96th Annual Meeting of the Japanese Urological Association. Yokohama, Japan. April 25-27, 2008. Abstracts]. PMID- 20017241 TI - [The 49th Annual Meeting of the Japanese Society of Neurology. Yokohama, Japan. May 15-17, 2008. Abstracts]. PMID- 20017242 TI - [Regional Meeting of the Japanese Radiological Society. 2006. Abstracts]. PMID- 20017243 TI - [The 70th Annual Meeting of the Japanese Society of Hematology. Kyoto, Japan. October 10-12, 2008. Abstracts]. PMID- 20017245 TI - [The 50th Congress of the Japanese Geriatric Society. Chiba, Japan. June 19-21, 2008. Abstracts]. PMID- 20017244 TI - Nursing leaders focus on future. PMID- 20017246 TI - [The 108th Annual Meeting of the Japanese Surgical Society. Nagasaki, Japan. May 15-17, 2008. Abstracts]. PMID- 20017247 TI - [The 81st Annual Meeting of the Japanese Society of Bacteriology, Kyoto, Japan. March 24-26, 2008. Abstracts]. PMID- 20017248 TI - [The 83rd Annual Meeting of the Japanese Society of Tuberculosis. Tokyo, Japan. April 24-25, 2008. Abstracts]. PMID- 20017249 TI - [The 50th Annual Meeting of the Japanese Society of Child Neurology. Tokyo, Japan. May 28-31, 2008. Abstracts]. PMID- 20017250 TI - [Abstracts of the 113th Annual Meeting of the Japanese Association of Anatomists. Oita, Japan. March 27-29, 2008. Abstracts]. PMID- 20017251 TI - [The 20th Spring meeting of the Japanese Society of Allergology. Tokyo, Japan. June 12-14, 2008. Abstracts]. PMID- 20017252 TI - [The 21st Spring Meeting of the Japanese Society of Allergology. Gifu, Japan. June 4-6, 2009. Abstracts]. PMID- 20017253 TI - [The 48th Annual Meeting of the Japanese Respiratory Society. Kobe, Japan. June 15-17, 2008. Abstracts]. PMID- 20017254 TI - Abstracts of the 9th Zagreb International Medical Summit (ZIMS) for Medical Students and Young Doctors. November 12-15, 2009. Zagreb, Croatia. PMID- 20017255 TI - Abstracts of the 8th Zagreb International Medical Summit (ZIMS) for Medical Students and Young Doctors. November 6-9, 2008. Zagreb, Croatia. PMID- 20017256 TI - [The 109th Annual Meeting of the Japanese Surgical Society. Fukuoka, Japan. April 2-4, 2009. Abstracts]. PMID- 20017257 TI - [The 67th Annual Meeting of the Japanese Radiological Society. Yokohama, Japan. April 4-6, 2008. Abstracts]. PMID- 20017258 TI - Freud's dreams of reason: the Kantian structure of psychoanalysis. AB - Freud (and later commentators) have failed to explain how the origins of psychoanalytical theory began with a positivist investment without recognizing a dual epistemological commitment: simply, Freud engaged positivism because he believed it generally equated with empiricism, which he valued, and he rejected "philosophy," and, more specifically, Kantianism, because of the associated transcendental qualities of its epistemology. But this simple dismissal belies a deep investment in Kant's formulation of human reason, in which rationality escapes natural cause and thereby bestows humans with cognitive and moral autonomy. Freud also segregated human rationality: he divided the mind between (1) an unconscious grounded in the biological and thus subject to its own laws, and (2) a faculty of autonomous reason, lodged in consciousness and free of natural forces to become the repository of interpretation and free will. Psychoanalysis thus rests upon a basic Kantian construction, whereby reason, through the aid of analytic techniques, provides a detached scrutiny of the natural world, i.e. the unconscious mental domain. Further, sovereign reason becomes the instrument of self-knowing in the pursuit of human perfection. Herein lies the philosophical foundation of psychoanalytic theory, a beguiling paradox in which natural cause and autonomous reason - determinism and freedom - are conjoined despite their apparent logical exclusion. PMID- 20017259 TI - [The 79th Annual Meeting of the Japanese Society of Hygiene. Tokyo, Japan. March 29- April 1, 2009. Abstracts]. PMID- 20017260 TI - [The 82nd Annual Meeting of the Japanese Society of Bacteriology. Nagoya, Japan. March 12-14, 2009. Abstracts]. PMID- 20017261 TI - [The 44th Autumn Meeting of the Japanese Radiological Society. Fukushima, Japan. October 22-24, 2008. Abstracts]. PMID- 20017262 TI - [The 106th Meeting of the Japanese Society of Internal Medicine. Tokyo, Japan. April 10-12, 2009. Abstracts]. PMID- 20017263 TI - [Abstracts of the 114th Annual Meeting of the Japanese Association of Anatomists. Okayama, Japan. March 28-30, 2009]. PMID- 20017264 TI - [The 58th General Meeting of the Japanese Society of Allergology. Tokyo, Japan. November 27-29, 2008. Abstracts]. PMID- 20017266 TI - [The ICF and its meaning for child and adolescent psychiatry]. PMID- 20017265 TI - Sociology and positivism in 19th-century France: the vicissitudes of the Societe de sociologie (1872-4). AB - Little is known about the world's first sociological society, Emile Littre's Societe de Sociologie (1872-4). This article, based on prosopographic research, offers an interpretation of the foundation, political-intellectual orientation and early demise of the society. As indicated by recruitment and texts by its founding members, the Societe de Sociologie was in fact conceived more as a political club than a learned society. Guided in this by Littre's heterodox positivism and the redefinition of sociology he proposed around 1870, the Societe de Sociologie was intended first and foremost to accompany intellectually the political changes that Littre considered imperative in the early years of the Third Republic (1870-1940). This expectation found little echo among the members of the society, and it seems possible that Littre himself and his closest associates were the ones to interrupt the society's meetings. Some of its members' general studies on the status of the social sciences and their main divisions were continued in the framework of the journal "La Philosophie positive" (1867-83), but the authors most committed to those studies were on the margins of the Littre network. Neither the dominant positivist republicanism, centered around Littre and Dubost, nor the general sociology of the more peripheral members of the network (Mesmer, Roberty, Vitry) represented an important intellectual contribution to the formation of academic sociology in France. Given that the Societe de Sociologie did contribute to diffuse the project of a sociological science and developed forms of sociology coherent enough to be rejected by the pioneers of university sociology, the group constitutes a significant case of failure in the history of the discipline. PMID- 20017267 TI - [On the trail of medical crimes: Keith Mant (1919-2000) and his debut as a forensic pathologist]. PMID- 20017268 TI - [Whole meal for the Fuhrer: the history of coarse wheat bread politics in the Third Reich]. PMID- 20017269 TI - [The industrial revolution of the sugar industry in Cuba, Brazil, and Argentina: technology and social change, ca. 1870-1930]. PMID- 20017270 TI - [Property, production, and labor in the southern Andean piedmont: the Zegada farms in the eastern valleys of Jujuy at the end of the colonial period]. PMID- 20017271 TI - Irish migrants in Paris hospitals, 1702-30: extracts from the registers of Bicetre, La Charite, La Pitie and La Salpetriere. PMID- 20017272 TI - [Forty years of papal discourse on health, 1939-78: the views of Pius XII, John XXIII, and Paul VI on the world of health]. PMID- 20017273 TI - Urfa and its environs in 1560s. PMID- 20017274 TI - The Brooke Hospital for animals. PMID- 20017275 TI - [The policy of population increase and the improvement of men: population discourse in the political and economic theory of the German Enlightenment]. PMID- 20017276 TI - Divorce and remarriage in Austria-Hungary: the second marriage of Franz Conrad von Hotzendorf. PMID- 20017277 TI - [The different faces of the regular garden in England, 17th-18th centuries]. PMID- 20017278 TI - [The great urban axes of 18th-century Warsaw and the creativity of Andre Lenotre]. PMID- 20017279 TI - [Tradition and modernization in Chinese marriage and family law]. PMID- 20017280 TI - [Social change in modern China and the development of women's education]. PMID- 20017281 TI - [The "Polish Versailles": the Bialystok of Jan Klemens Branicki]. PMID- 20017282 TI - [Drug counterfeiting and counter measures. Study by the Hungarian National Alliance Against Drug Counterfeiting]. PMID- 20017283 TI - Treatment of lymphoblastic lymphoma in adults. AB - Lymphoblastic lymphoma is a rare disease in adults, primarily affecting patients in their late teens and early 20s. Optimal treatment strategies have been slow to emerge because of the rarity of this disease and the variable distinction in the clinical literature between this condition and acute lymphoblastic leukemia. Although these two conditions are now regarded as a single entity in the WHO Classification of Lymphoid Neoplasms, treatment approaches have developed separately, and recent molecular data suggest that there may be important biologic differences between these conditions that may justify a different treatment approach. Most published data support the use of intensive multiagent chemotherapy induction followed by a consolidation and maintenance phase. Optimal consolidation remains unclear, although there is no clear role of stem cell transplantation after intensive remission induction therapy based on current evidence. Emerging molecular data have identified potential new therapeutic targets with supporting preclinical data. PMID- 20017284 TI - Further considerations about lymphoblastic lymphoma. PMID- 20017285 TI - Diagnosing and treating chronic lymphocytic leukemia in 2009. AB - Over the past decade, major breakthroughs have been made in both the molecular understanding and the treatment of chronic lymphocytic leukemia (CLL). In this article, old and new concepts of CLL biology are explored and insights into the relevance of the newer prognostic factors are discussed. The therapeutic landscape has changed dramatically with the advent of purine analogs, monoclonal antibodies, and combination therapy. As opposed to older agents, these new therapies commonly yield complete remissions. This improvement has spurred a debate as to new goals in treating CLL patients. PMID- 20017286 TI - Interesting times in the diagnosis and treatment of CLL. PMID- 20017287 TI - New menus for CLL treatment. PMID- 20017288 TI - Managing colorectal cancer liver metastases. AB - The treatment of resectable colorectal cancer metastases to the liver has undergone changes with the addition of active chemotherapeutic agents, innovations and definition in the surgical procedures, understanding of the benefits and toxicities of the surgical and chemotherapeutic (cytotoxic and biologic) interventions, and use of the team approach. Patients are initially evaluated for the overall risk of their disease, which includes the standard parameters for disease recurrence and blends in disease and patient comorbidities and likelihood of surgical success. Advanced imaging techniques are mandatory in the initial evaluation. Rather than approaching the patient with sequential, independent therapies and handoff from specialist to specialist, a continuous interaction is required. This article outlines the initial consultation, required team components, surgical decision-making, and use of cytotoxic and biologic agents. The implication is that the best outcomes can only be achieved with the use of all modalities. PMID- 20017289 TI - The multidisciplinary approach to colorectal cancer liver metastases. PMID- 20017290 TI - Changing management strategies for hepatic colorectal metastasis. PMID- 20017291 TI - Intensity-modulated radiation therapy for anal cancer. AB - The contemporary treatment of anal cancer is combined-modality therapy with radiation therapy, fluorouracil, and mitomycin. This therapy results in long-term disease-free survival and sphincter preservation in the majority of patients. Tempering these positive results is the high rate of treatment-related morbidity associated with chemoradiation therapy for anal cancer. The use of intensity modulated radiation therapy (IMRT) has the potential to reduce acute and chronic treatment-related toxicity, minimize treatment breaks, and potentially improve disease-related outcomes by permitting radiation dose escalation in selected cases. PMID- 20017292 TI - Jury still out on whether advanced technology can improve the outcomes of patients with anal canal cancer. PMID- 20017293 TI - Advanced radiation technology in the treatment of anal cancer. PMID- 20017294 TI - Hormone replacement and breast cancer risk: reconsidering the data. PMID- 20017295 TI - Complementary therapies, herbs, and other OTC agents: Boswellia (Boswellia serrata). PMID- 20017296 TI - [A novel mutation of the SLC34A2 gene in a Chinese pedigree with pulmonary alveolar microlithiasis]. AB - OBJECTIVE: To identify the mutation of solute carrier family 34 member 2 (SLC34A2) gene in a Chinese family with pulmonary alveolar microlithiasis (PAM). METHODS: Genomic DNA was extracted from the family members. DNA sequencing was carried out to confirm the mutation detected by polymerase chain reaction-single strand conformation polymorphisms (PCR-SSCP). The fragments with variation were screened in 100 healthy controls by PCR-SSCP. RESULTS: In both patients of the family, a homozygous mutation of the SLC34A2 gene was identified in exon 8 (c.A910T), resulting in a premature stop codon. In addition, a homozygous single nucleotide polymorphism (SNP) was found in intron 2 in both patients and the daughter of proband. CONCLUSION: A novel homozygous mutation in SLC34A2 gene, leading to a premature stop codon therefore a truncated protein, was probably responsible for the PAM in this family. The SNP in intron 2 needs further study. PMID- 20017297 TI - [Detection of microdeletion in Williams syndrome by multiplex ligation-dependent probe amplification]. AB - OBJECTIVE: To establish a method of multiplex ligation-dependent probe amplification (MLPA) for clinical screening of Williams syndrome (WS) and for routine use in WS diagnosis. METHODS: Probes for MLPA were designed according to the frequent deletion regions, and used to screen the two patients suspected with Williams syndrome, and the density of the bands were analyzed with software. Linkage analysis using polymorphic markers was performed to confirm the positive result of MLPA. RESULTS: The MLPA data indicated that the two children had possible microdeletions in the WS critical region. The deletions were confirmed and both were maternal origin by polymorphism analysis. CONCLUSION: MLPA is a quick and convenient method for detecting deletion or duplication mutations. It can provide reliable and helpful information for clinical diagnose of Williams syndrome. PMID- 20017298 TI - [Establishment of immortalized cell lines and genetic stability evaluation for a family with spinocerebellar ataxia type 2]. AB - OBJECTIVE: Immortalized cell lines of spinocerebellar ataxia type 2 (SCA2) with Parkinson disease symptoms were established in order to provide experimental material for future study. METHODS: The immortalized cell lines were constructed by using Epstein Barr virus and cyclosporine A. Microsatellite markers were detected to see whether there is any change between the cell lines and the original blood samples, and the genetic stability of the cell lines were evaluated. RESULTS: Twenty-five immortalized cell lines were established successfully from the family and the microsatellite markers were unchanged. CONCLUSION: The karyotypes of the immortal cell lines were normal and the cell lines were genetically stable. PMID- 20017299 TI - [DRB1 * 1454: the common allele of human leukocyte antigen-DRB1 * 14 in Chinese Han populations]. AB - OBJECTIVE: To investigate and compare the distribution of HLA-DRB1 * 14 alleles between the southern and northern Chinese Han populations. METHODS: Human leukocyte antigen (HLA)-DRB1 alleles of 436 southern and 713 northern Chinese Han bone marrow volunteers were genotyped by polymerase chain reaction (PCR)-sequence based-typing (SBT) method, among them the DRB1 * 1401/1439/1454 ambiguous allele pairs were identified using DRB1 * 14 high-resolution PCR-sequence specific primer (SSP) kits. Also, the clinic samples previously reported as DRB1 * 1401 were re-genotyped using the same PCR-SSP kits. The allelic distribution of DRB1 * 14 in southern and northern Chinese Han population were compared by chi-square test method. RESULTS: Eighty-one ambiguous allele pairs of DRB1 * 1401/1439/1454 and 54 clinic samples previously reported as DRB1 * 1401 were all identified as DRB1 * 1454. Among the 436 Southern Han donors, six subtypes of DRB1 * 14 allele were observed including DRB1 * 1454 (69.57%), DRB1 * 1402 (1.45%), DRB1* 1403 (1.45%), DRB1 * 1404 (4.35%), DRB1 * 1405 (20.29%) and DRB1 * 1407 (2.90%). In the 713 northern Han donors, a total of seven subtypes were observed including DRB1 * 1454 (35.48%), DRB1 * 1403 (12.90%), DRB1 * 1404 (6.45%), DRB1 * 1405 (37.63%), DRB1 * 1407 (4.30%), DRB1 * 1411 (1.08%) and DRB1 * 1412 (2.15%). CONCLUSION: DRB1 * 1454 and DRB1 * 1405 were the most common alleles of HLA-DRB1 * 14 in Chinese Han populations. The distribution of HLA-DRB1 * 14 differ significantly between the southern and northern Chinese Han population, while DRB1 * 1405 showed similar distribution pattern in the two populations but DRB1 * 1454 had higher frequency in southern than in northern Chinese Han population. PMID- 20017300 TI - [Construction of recombinant adenovirus vector expressing shRNAs targeting COX-2, AKT1 and PIK3R1 gene and its inhibition effect on proliferation of human gastric adenocarcinoma]. AB - OBJECTIVE: To construct a recombinant adenovirus vector that expresses small hairpin RNAs (shRNA) against COX-2, AKT1 and PIK3R1 gene and to evaluate its potential for suppressing the cell proliferation of human gastric adenocarcinoma SGC701 cell in vitro and in vivo, which will enable the development of a gene therapy protocol for the treatment of human gastric adenocarcinoma. METHODS: Three strips of shRNA targeting AKT1, COX-2 and PIK3R1, was subcloned into adenovirus expression vector. After verification, it was amplified and titered. The recombinant adenovirus expression vector was infected into human gastric adenocarcinoma SGC7901 cells in vitro and the infected cells were injected in nude mice. The mRNA and protein expression levels of AKT1, COX-2 and PIK3R1 were determined by real-time PCR and Western blot respectively. Cell proliferation in vitro was determined by methyl thiazolyltetrazolium (MTT) assay and flow cytometry, tumor growth in vivo was measured by volume of tumor in nude mice. RESULTS: Restriction digestion and sequencing analysis showed that the rAd5-C-A-P adenovirus expression vector was constructed successfully. It significantly inhibited the expression of AKT1, COX-2 and PIK3R1, and cell growth was inhibited over 70% as indicated by MTT assay and accompanied with G0/G1 phase arrest. Cell growth on matrigel matrix showed that the rAd5-C-A-P transfected cells were detached from the matrix or grew in a scattered clustering pattern, indicating poor cell growth activities in 2-D matrigel. Tumor growth in nude mice in the C + A + P group was inhibited (P<0.01). CONCLUSION: shRNA targeting COX-2, AKT1 and PIK3R1 down regulated significantly the expression of the three genes in a sequence-specific manner, exerted proliferation inhibition effect on SGC7901 cells in vitro and in vivo. PMID- 20017301 TI - [Association of serotonin and norepinephrine transporter gene polymorphisms with the susceptibility to depression]. AB - OBJECTIVE: To determine whether the serotonin transporter (5-HTT) and norepinephrine transporter (NET) gene polymorphisms were associated with the susceptibility to depression. METHODS: Five hundred and seventy-nine patients with depression, evaluated using a 17-item Hamilton Depression Rating Scale for Depression (HAMD), and 437 healthy controls, all of Chinese Han origin, were genotyped by polymerase chain reaction. RESULTS: Both genotype distributions (P=0.033) and allele frequencies (P=0.023, OR = 1.250, 95% CI = 1.031-1.517) of NET-T182C were significantly different between patients and controls, where the T allele was associated with the onset of depression. Both NET-T182C T- and 5- HTTLPR L-carriers had higher baseline HAMD scores (P=0.032 and 0.023, respectively). There was an interaction between NET-T182C and 5-HTTLPR, where the combined genotype distributions were associated with both onset of depression (P=0.006) and the baseline HAMD scores (P=0.007). CONCLUSION: This study suggested a positive relationship between the NET-T182C polymorphism and the susceptibility to depression, and a positive relationship between NET-T182C/5 HTTLPR polymorphisms and the severity of depression. PMID- 20017302 TI - [Application studies on the gene diagnosis and carrier detection of hemophilia A by using polymerase chain reaction-conformation sensitive gel electrophoresis]. AB - OBJECTIVE: To establish a simple, rapid and easy method for screening the gene mutation in hemophilia A, which was further applied to a direct diagnosis and carrier detection at gene level. METHODS: Twenty-four clinically diagnosed hemophilia pedigrees, including all the hemophilia patients and female members, were tested for the introns 22 and 1 in factor VIII gene by using inversion polymerase chain reaction (PCR) and regular PCR techniques. All the 26 exons of factor VIII gene were consecutively screened in the 17 patients manifesting non inverted sequences in intron 22 by using PCR, subsequently all the 37 amplicons resulted from 26 exons were analyzed by conformation sensitive gel electrophoresis (CSGE), finally the mutated exons were subjected to sequencing verification. According to the mutation results, mothers and twin sisters of the hemophilia probands were tested by CSGE or subjected to nucleotide sequencing directly, to ascertain if those individuals had the same mutation or were the carriers of disease-causing gene. RESULTS: Intron 22 inversion was detected in 7 hemophilia probands out of 24 hemophilia pedigrees, intron 1 inversion was not detected in these pedigrees. Single-base mutations distributed in different exons of factor VIII gene were detected in 13 pedigrees with family history and 3 sporadic pedigrees, diagnosed as non-inverted 22 intron patients. By comprehensive usage of PCR-CSGE and nucleotide sequencing, the positive rate and the diagnosable rate of gene diagnosis or carrier detection in the 24 hemophilia pedigrees was 94.12% and 100% respectively. CONCLUSION: PCR-CSGE is a highly sensitive and special assay for detecting single base mutation. By integrated utilization of introns 22 and 1 of factor VIII gene detection and PCR-CSGE genotyping, combining with nucleotide sequencing, a direct diagnosis of all hemophilia pedigrees be could nearly make at gene level, including the sporadic families. This method might be used to screen new mutation theoretically and ascertain the mutation type. It is a simple, rapid and low-cost method, possessing unique advantages in direct diagnosis of hemophilia A and carrier screening. It should have important application value in hemophilia diagnosis. PMID- 20017303 TI - [Effect of YB-1 gene knockdown on human leukemia cell line K562/A02]. AB - OBJECTIVE: To investigate the potential effects of YB-1 gene knockdown on gene expression profile, cell growth and apoptosis in leukemia cell line K562/A02. METHODS: The recombinant eukaryotic expression plasmid containing YB-1 short hairpin RNA (shRNA) or random-sequence (HK) were transfected into K562/A02 cells by lipofectamine mediation. cDNA microarray was performed to explore the alteration of gene expression profile when YB-1 gene expression was decreased. Expression of CARD8 and RHOC genes were verified by semi-quantitative reverse transcription-PCR (RT-PCR). The proliferative ability of the cells was determined by methyl thiazolyltetrazolium (MTT) assay and cell cycle analysis. Cell apoptosis was assayed by Annexin V-FITC/PI double labeled flow cytometry. RESULTS: The levels of YB-1 mRNA and protein decreased dramatically in three positively transfected cells when compared with untransfected K562/A02 cells or K562/A02-HK thansfected cells. Gene expression profile was altered by transfection of YB-1 shRNA into K562/A02 cells. Among 47,000 genes on the microarray, 252 genes were detected to have changes, with 143 down-regulated and 109 up-regulated. They were functionally related to cell cycle progression, gene replication, metabolism, cell apoptosis, cell signal transduction, etc. An increase in CARD8 gene expression and a decrease in RHOC gene expression have been confirmed by RT-PCR in K562/A02-YBX13 cells. The introduction of exogenous YB-1 shRNA gene into K562/A02 cells resulted in decreased proliferation, higher G1, lower G2 and S ratio in cell cycle distribution in comparison with the control groups. Annexin V/PI detection indicated higher Annexin V+ ratio in the three positively transfected cells 24 hours after cells were treated with 0.5 micromol/L of As2O3. CONCLUSION: Down-regulation of YB-1 gene by shRNA-YB-1 can alter the gene expression profile in K562/A02 cells, leading to change of cell proliferation and apoptosis. PMID- 20017304 TI - [Detection of the CAG trinucleotide repeats of MJD1 gene by recombinant DNA technology]. AB - OBJECTIVE: To establish a stable, accurate and intuitive method for detecting the CAG trinucleotide repeats of MJD1 gene. METHODS: The CAG trinucleotide polymorphism of the MJD1 gene was analyzed by recombinant DNA technology and DNA sequencing in 35 spinocerebellar ataxia 3/Machado-Joseph disease (SCA3/MJD) patients from Mainland China. RESULTS: The range of the CAG repeat of the 35 patients was 65-81 (mean = 72.96 +/- 4.24). The CAG repeats contained two CAAs and one AAG variations in the CAG motif in all the patients and majority of the healthy controls. There was a CGG/GGG polymorphism at the 3' end of the CAG repeat. The GGG allele was consistently associated with smaller CAG repeats in healthy controls. On the other hand, the CGG allele consistently existed in the patients. CONCLUSION: Recombinant DNA technology can stably, accurately and intuitively detect the CAG trinucleotide repeat of the MJD1 gene. It should be used as a major technique to diagnose the SCA3/MJD and analyze the polymorphism of CAG sequence. PMID- 20017305 TI - [Noninvasive prenatal diagnosis of single gene disorders through cell-free fetal DNA in maternal blood]. AB - The discovery of cell-free fetal DNA (cff-DNA) in maternal plasma offered a new way to noninvasive prenatal diagnosis for single gene disorders. In the past decade, many techniques such as real-time PCR, pyrophosphorolysis-activated polymerization, mass spectrum and digital PCR have been developed for noninvasive prenatal diagnosis. In this review, the author discuss the principles, applications, advantages and disadvantages of these techniques. PMID- 20017306 TI - [Advances in molecular genetic study of myopia]. AB - Myopia is an important cause of blindness, in which an image is focused in front of the retina. Genetic factors have been implicated in the pathogenesis of myopia. Based on the molecular genetic study, some genetic loci linked to myopia have been mapped, but no disease-causing gene has been identified. Here authors review the genetic study on myopia, including gene mapping and candidate gene screening. PMID- 20017307 TI - [Mutation analysis of the PAH gene in patients with phenylketonuria in Gansu province]. AB - OBJECTIVE: To characterize the mutations of the phenylalanine hydroxylase (PAH) gene in patients with phenylketonuria in Gansu province. METHODS: Mutations of the PAH gene were detected in exons 3, 5, 6, 7, 11 and 12 with flaking introns of PAH gene by PCR and DNA sequencing. RESULTS: Mutations were identified in 45/58 alleles (detection rate: 96.4%), in total of 18 variants. Among them IVS12+5G>C was a novel mutation. The most frequent mutations were R243Q (22.7%), V399V (12.1%), EX6-96A>G (5.2%), R413P (5.2%) and IVS4-1G>A (5.2%), followed by Y356X (3.4%), R111X (3.4%) and INS7+2T>A (3.4%). CONCLUSION: The mutations of the phenylalanine hydroxylase gene in patients with phenylketonuria in Gansu province were similar to that in other areas of China, with obvious difference in mutation rate of some mutations. PMID- 20017308 TI - [Association of IL-10 gene polymorphisms with gastroduodenal diseases in Hubei Han population]. AB - OBJECTIVE: To study the distribution of IL-10 gene polymorphisms in patients with gastroduodenal diseases in Hubei Han population and their association with helicobacter pylori (Hp) infection. METHODS: Six hundred and five patients with gastroduodenal diseases (220 gastric cancer, 196 chronic gastritis and 189 gastroduodenal ulcer) and 624 healthy controls were genotyped by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method for IL-10 1082, -819, -592 gene polymorphisms. Hp infection status was determined by ELISA. RESULTS: (1) There was significant difference of IL-10 -1082 AG+GG genotypes between gastric cancer group and gastric cancer-free and healthy control groups (P<0.05). (2) There was no significant difference of IL-10 -592 and IL-10 -819 gene polymorphisms among gastric cancer, gastric cancer-free and healthy control groups (P>0.05). (3) The frequency of IL-10 -1082 AG+GG genotypes in the Hp positive gastric cancer patients was significantly higher than that of control groups (P<0.05). CONCLUSIONS: (1) Genotypes AG+GG of IL-10 -1082 were associated with gastric cancer in Hubei Han population. (2) The IL-10 -1082 AG+GG genotypes were associated with Hp infection in patients with gastric cancer. PMID- 20017309 TI - [Genetic defect in Chinese azoospermic patients and their relationship with reproductive hormones]. AB - OBJECTIVE: To investigate the incidence of abnormal karyotypes and Y chromosome microdeletion in Chinese men with azoospermia, and the relationship with reproductive hormones. METHODS: Four hundred and eighty nine cases of azoospermic patients and 20 fertile men were studied. Karyotypes and Y chromosome microdeletion were analyzed by G-banding and mutiplex polymerase chain reaction, respectively. Chemiluminescene immunoassay technique was applied to measure the serum levels of follicle-stimulating hormone (FSH), luteinizing hormone (LH), testosterone (T), and prolactine (PRL). RESULTS: Chromosome abnormalities were found in 102 out of 489 azoospermic patients (20.86%), among them 86 (84.31%) cases had sex chromosome abnormalities, with 73 cases being Klinefelter syndrome. Y chromosome microdeletions were detected in 58 (11.86%) cases out of the 489 patients, and deletion of the AZFc region was the leading group (63.8% of all deletions), followed by AZFbc (19.0%), AZFabc (10.3%), AZFb or AZFa (3.4%). FSH, LH levels were significantly increased and T level was decreased in azoospermic patients compared with the fertile men group (P<0.01). Furthermore, in the azoospermic patients with Klinefelter syndrome or AZFabc microdeletions, FSH and LH levels were increased more significantly, and were statistically different from azoospermic patients with normal karotype or without Y chromosome microdeletion (P<0.05). CONCLUSION: In the Chinese men with azoospermia, the incidence of abnormal karyotype and Y chromosome microdeletion were similar to those described previously in other populations. In azoospermia with Klinefelter syndrome or AZFabc microdeletions, FSH and LH levels increased markedly indicating the protracted stimulation of gonadotrophs due to lack of androgen feedback. PMID- 20017310 TI - [Association of cytotoxic T lymphocyte antigen-4 promoter C-658T polymorphism with ulcerative colitis in Chinese]. AB - OBJECTIVE: To investigate the association of gene polymorphism of cytotoxic T lymphocyte-associated antigen 4 (CTLA-4) with ulcerative colitis (UC) in Chinese. METHODS: One hundred and seventeen patients with UC and 246 healthy controls were genotyped for the polymorphisms of C-658T in the promoter and C61T at the 3' untranslated region of the CTLA-4 gene using polymerase chain reaction restriction fragment length polymorphism (PCR-RFLP) and polymerase chain reaction single strand conformation polymorphism (PCR-SSCP), respectively. The genotype and allele frequencies of the two groups were calculated and compared by chi square test. RESULTS: The frequency of TT+CT genotype at the CTLA-4 gene C-658T in the promoter was significantly higher in UC patients than that in healthy controls (P=0.015). The frequency of the T allele at this locus was also significantly higher in UC patients than that in the controls (P=0.033). The frequencies of TT genotype and T allele at the C-658T locus were highly associated with extensive colitis in UC patients (P=0.037, and P=0.0067, respectively). CONCLUSION: The T allele of CTLA-4 promoter C-658T locus was highly associated with UC in Chinese Han of central China. PMID- 20017311 TI - [Study on the association of the CRP gene +1444C/T polymorphism with symptomatic carotid artery stenosis]. AB - OBJECTIVE: To investigate the potential association of the C-reactive protein (CRP) gene +1444C/T polymorphism with symptomatic carotid artery stenosis. METHODS: Polymerase chain reaction-restriction fragment length polymorphism was used for the detection of CRP +1444C/T genotypes in 192 patients with symptomatic carotid artery stenosis and 197 healthy controls. Serum high sensitivity-CRP (hs CRP) levels were measured by routine method. RESULTS: No TT genotype was detected in this study. Patients with >70% stenosis had higher CC genotype compared with those with <70% stenosis after adjusting for major cerebrovascular risk factors (OR: 2.958; 95% CI: 1.198 - 7.305; P=0.019). CRP levels were significantly higher in patients than in controls. Subgroup analysis according to clinical characteristics (single or double stenosis; >70% or <70% stenosis) did not show difference in CRP levels. There was no significant difference in the prevalence of CT genotype between patients and controls, or between single and double stenosis (P>0.05). CONCLUSION: The CRP +1444 CC genotype is a risk factor for >70% carotid artery stenosis. The serum CRP level is associated with the presence of carotid stenosis. However, it is not associated with the number and severity of stenosis. PMID- 20017312 TI - [Application of multicolor primed in situ labeling in simultaneous identification of chromosomes 18, X and Y in uncultured amniocytes]. AB - OBJECTIVE: To establish a multicolor primed in situ labeling (PRINS) protocol for chromosome detection in uncultured amniocytes. METHODS: Chromosomes 18, X and Y in uncultured amniocytes were simultaneously detected by using the non-ddNTP blocking multicolor PRINS procedure. RESULTS: Within 7 h, the 3 chromosomes were simultaneously marked in the same uncultured amniocyte. The chromosome signals were successfully detected in 69 uncultured samples of amniotic fluid. The results were consistent with that obtained by chromosomes in cultured amniocytes. CONCLUSION: This multicolor protocol was high throughput, fast, simple, sensitive and reliable in diagnosing chromosome abnormalities in uncultured amniocytes. PMID- 20017313 TI - [Trisomy 8 in chronic lymphocytic leukemia]. AB - OBJECTIVE: To investigate the incidence of trisomy 8 in chronic lymphocytic leukemia (CLL) and its significance in prognosis. METHODS: A panel of probes and fluorescence in situ hybridization (FISH) were used to detect trisomy 8 in 151 CLL patients combined with chromosome karyotype analysis. RESULTS: There were 2 patients (1.3%) with trisomy 8 in the 151 CLL patients, and the number of trisomy 8 cells was 8% and 10% respectively. The karyotypes were 47,XY,+8[2]/49,XY,+14,+20,+21[2]/ 46,XY[16], and 47,XX,+8[2]/46,XX[18], respectively. CONCLUSION: Trisomy 8 was rare in CLL, and its significance in prognosis of CLL still remains unknown. PMID- 20017314 TI - [Study on the differentially expressed genes of steroid-resistant and steroid sensitive nephrotic syndrome]. AB - OBJECTIVE: To identify the genes differentially expressed among the steroid resistant nephrotic syndrome (SRNS), steroid-sensitive nephrotic syndrome (SSNS) and normal children, and understand the molecular mechanism of SRNS. METHODS: Affymetrix microarray technology was used to obtain such a profile. The differentially expressed genes among these groups were identified based on signal to-noise ratios by GCOS software; real-time PCR analysis was performed to confirm the microarray results. RESULTS: There were 157 genes differentially expressed among these groups. The genes up-regulated both in SRNS and SSNS were involved primarily in ionic transportation, immuno-signal transduction and apoptosis. In particular, CLNS1A gene was down regulated in SRNS but up regulated in SSNS. CONCLUSION: Several differentially expressed genes, such as CLNS1A and HLA-DRB4 were found to be closely related to the pathogenesis of SRNS and SSNS. This DNA microarray analysis has provided some important clue to the molecular mechanism of SRNS. PMID- 20017315 TI - [Association of the D358A polymorphism of IL6R gene with type 2 diabetes in Hubei Han Chinese]. AB - OBJECTIVE: To investigate the association of the D358A polymorphism of interleukin 6 receptor( IL6R ) gene with type 2 diabetes mellitus (T2DM) in Hubei Han Chinese. METHODS: The single nucleotide polymorphism D358A of the IL6R gene was genotyped in 581 T2DM patients and 353 healthy controls. Meta-analysis was used to assess the reported association between the IL6R D358A and T2DM. RESULTS: The frequencies of CC genotype and C allele of IL6R D358A in the patients were significantly lower than those in controls (CC: 13.4% vs. 20.7%, P=0.003; C: 36.0% vs. 41.8%, P=0.012), with the CC genotype being a protective factor for T2DM (OR=0.595, P=0.003). Logistic regression analysis suggested that the CC genotype was significantly associated with T2DM after adjusted for age, sex, blood pressure and obesity (OR=0.615, 95% CI: 0.407- 0.928, P=0.021). Meta analysis of 6 studies indicated that there existed genetic heterogeneity, and the CC genotype was associated with lower risk of T2DM (P=0.009, OR=0.64, 95% CI: 0.48-0.85). CONCLUSION: IL6R is a susceptibility gene for T2DM in Han Chinese population of Hubei, and the CC genotype may serve as a genetic protective factor of T2DM. PMID- 20017316 TI - [Studies on the mutation and polymorphism of the TPMT gene in Chinese children with acute leukemia]. AB - OBJECTIVE: To investigate the allelic frequencies and distribution of single nucleotide polymorphisms within the coding region (cSNPs) of thiopurine S methyltransferase gene (TPMT) in Chinese children with acute leukemia (AL) and healthy controls, in order to provide genetic references for individual chemotherapy for AL patients by studying the relationship between the cSNP in human TPMT and chemotherapeutic effect of thiopurine drugs. METHODS: The bone marrow samples from 53 children with AL and peripheral blood samples from 115 healthy children were obtained to prepare complementary DNAs (cDNAs). The cDNAs were analyzed for the polymorphisms in the TPMT gene by reverse transcriptase polymerase chain reaction (RT-PCR)-denaturing gradient gel electrophoresis (DGGE) and DNA sequencing. The distribution of each genotype was evaluated. RESULTS: Two novel heterozygote mutations, 210C>T (C70C, silent) and 622T>C (F208L), were identified in the coding region of the TPMT in a single sample, respectively. The mother of the child with mutation 622T>C was confirmed as the same genotype by DGGE and sequencing (NCBI_ss accession numbers 107796292 and 107795933). Two known polymorphisms, 474T>C (silent) and 719A>G (T240C), were identified. The allelic frequencies were 14.2%, 2.83% and 17.0%, 3.04% in the AL children and control children respectively, with the total allelic frequencies of 16.2% (first reported in the Chinese Han population) and 2.99% respectively. No association with susceptibility to disease was observed. CONCLUSION: Two novel mutations and two known polymorphisms were identified in Chinese children by RT-PCR-DGGE combined with DNA sequencing, which provides the first step to identify genetic markers for predicting variability in response to and toxicity of thiopurine drugs. PMID- 20017317 TI - [Analysis of the small supernumerary marker chromosome in Turner syndrome with 45, X/46, X, + mar karyotype]. AB - OBJECTIVE: To identify the origin and study the morphology of small supernumerary marker chromosome (sSMC) in Turner syndrome with 45, X/46, X, + mar karyotype. METHODS: Using the conventional chromosome G-banding technique, 10 cases of Turner syndrome with 45, X/46, X, + mar chromosome karyotype were obtained, dual color fluorescence in situ hybridization was applied to study the origin and morphology of the sSMC. RESULTS: In the 10 cases of Turner syndrome with 45, X/46, X, + mar karyotype, the sSMC of 7 cases was derived from X chromosome [sSMC(X)], the sSMC of 2 cases was derived from Y chromosome [sSMC(Y)] and the remaining 1 case was derived from the autosome. There were 4 cases of ring(r) chromosomes and 3 of centric minutes (min) in the 7 sSMC (X) cases. In the 2 sSMC(Y), one case was dicentric (dic) and the other was centric minute (min). The sSMC originated from the autosome was a centric minute (min). CONCLUSION: The origin of sSMC of Turner syndrome with 45, X/46, X, + mar karyotype was almost all from sex chromosomes, and rarely from autosomes. sSMC can exist as isodicentric, ring, or centric minute. The molecular cytogenetic features of the sSMC can provide useful information for genetic counseling, prenatal diagnosis and treatment of the Turner syndrome patients with a 45, X/46, X, + mar karyotype. PMID- 20017318 TI - [Genetic polymorphisms of fifteen short tandem repeat loci in Guilin Han population]. AB - OBJECTIVE: To investigate the genetic polymorphisms of 15 short tandem repeat (STRs) in Guilin Han population. METHODS: DNA was extracted by Chelex method, and 15 STRs were analyzed using AmpFISTR Identifiler kit. RESULTS: Four rare alleles, namely FGA * 10, D2S1338 * 10, D3S1358 * 16.2 and D3S1358 * 17.2, were observed. The combined match probability and exclusion probability for the 15 STRs were 2.89 x 10(-17) and 0.9999993, respectively. CONCLUSION: These STRs have good discrimination power and exclusion probability in Guilin Han population. PMID- 20017319 TI - [Polymorphism of fifteen short tandem repeat loci in Tibetan of Changdu area]. AB - OBJECTIVE: To investigate the genetic polymorphism of 15 autosomal short tandem repeat (STR) loci, i.e. D5S818, FGA, D8S1179, D21S11, D7S820, CSF1PO, D3S1358, TH01, D13S317, D16S539, D2S1338, D19S433, vWA, TPOX and D18S51, in Tibetan population of Changdu area, and to analyze the genetic relationship between this population and other Asian populations. METHODS: The 15 loci were amplified simultaneously using a multiplex PCR typing system. Amplified STR fragments were analyzed with an ABI PRISM 3100 Avant DNA Analyzer. Arlequin software version 3.1 was used to obtain allele frequencies and polymorphism parameters. RESULTS: There were 135 STR alleles in the 15 STRs of Changdu Tibetan, with allele frequencies ranging from 0.0065 to 0.5455. The average heterozygosity was 0.7340, the discrimination power was higher than 0.8 except for TPOX and TH01. The accumulative discrimination power was more than 0.9999998, and the accumulative non-parental exclusion probability was 0.99999997. CONCLUSION: The 15 STR loci of Changdu Tibetan possess characteristics of high genetic diversity. They can be employed in minority genetics investigation, and individual and paternity test in forensic medicine. PMID- 20017320 TI - [Blazing new areas of microinvasive arthroscopic surgery]. PMID- 20017321 TI - [Anatomic and clinical study on 4-bone-tunnel double-bundle anterior cruciate ligament]. AB - OBJECTIVE: To conduct a primary clinical analysis on 411 cases of arthroscopic 4 bone-tunnel double-bundle anterior cruciate ligament (ACL) reconstruction done by one operator on the basis of anatomic measurements of cadaver knees. METHODS: Twenty-two adult male Chinese cadaver knees of 25-45 years old were dissected. The sizes, distributions and positions of femoral and tibia footprints of anterior medial bundle (AMB) and posterior lateral bundle (PLB) of ACL and their fiber's orientations were measured and recorded. The anatomy and measurement results were used by one operator in 413 double-bundle ACL reconstruction knees of 411 cases with arthroscopic 4 bone-tunnel technique. Among them, 297 males and 114 females, there were 222 left knees and 191 right knees. The average age was 27.61 +/- 7.23 years old. RESULTS: The anatomy results showed the locations and sizes of AMB and PLB footprints in both femoral and tibia sides were different for different individuals. It was suggested to locate AMB femoral tunnel 7.95 +/- 1.40 mm inferior to 12:00 o'clock position on the posterior edge of the lateral wall of intercondylar fossa with 1 mm thick of the posterior wall of finished AMB femoral tunnel. With the knee flexed to 90 degrees, the PLB tunnel was 5.05 +/- 0.76 mm superior to the lower cartilage edge of the lateral wall of intercondylar fossa, 8.60 +/- 1.52 mm away from the anterior cartilage edge and 8.65 +/- 1.54 mm from the posterior edge. The anatomic orientations of tibia and femoral tunnels were also measured. The study results were used in arthroscopic double bundle ACL reconstructions of 413 knees. In double-bundle ACL reconstruction operations, the grafts were used in the way from 2-strip double-bundle to 9-strip double-bundle. For most cases, Endo-Button was used for femoral fixation and bio absorbable interference screw and staple were used for tibia fixation. For the patients adopting autologous semitendinosus and gracilis tendons (STG), the diameter of PLB bone tunnels was 5-6 mm in 95.9% cases, while that of AMB bone tunnel 5-7 mm in 72.4% cases. After a 28-month follow-up in 75 cases, the results showed that double-bundle ACL reconstruction could better reconstruct the stability of knees. CONCLUSION: In 4-bone-tunnel double-bundle arthroscopic ACL reconstruction, both bone tunnel positions and their orientations should be determined according to the anatomic measurement results. Since the PLB diameters of 95.9% cases were 5-6mm, considering the operative outcome of ACL revision, the double bundle ACL reconstruction technique was safe. A better knee stability could be reconstructed by this technique. PMID- 20017322 TI - [Gait analysis associated with anterior cruciate ligament reconstruction]. AB - OBJECTIVE: To determine how selected gait parameters, measured by three dimensional To determine how selected gait parameters, measured by three dimensional motion capture system, may change as a result of anterior cruciate ligament (ACL) deficiency and following ACL reconstruction. METHODS: The study was performed on 29 ACL-deficient subjects prior to and at 3, 6, 9 and 12 months after ACL reconstructive surgery by the four stranded hamstring tendon technique. The group was examined at an average of 2.6 months after injury (range: 2 weeks 16 months). But at presurgery, gait analysis was performed using the DorealSoft DVMC-8801 three-dimensional motion capture system. Kinematic data were recorded for the lower extremities. The results obtained from the injured subjects were compared with those of 58 individuals without ACL damage. Each subject was asked to walk on a motorized treadmill (ECON USA) with different slopes and perform after a 6 minute familiarization time-at least 2 minutes of walking at a constant speed of 1.9 km/h. RESULTS: The ACL-deficient patients exhibited a quadriceps avoidance pattern prior to and at 3 months post-surgery. In the operated individuals, the spatial-temporal parameters and the knee angle had already regained a normal pattern for the ACL-deficient extremity during gait at 6 months post-surgery. However, the relative ACL movement parameter-which describes the tibial translation into the direction of ACL-and the angular acceleration showed no significant statistical difference as compared with the values of healthy control group at just 9 months post-surgery. CONCLUSION: The ACL surgical repair significantly alters lower-extremity gait patterns, and that the establishment of pre-injury gait patterns takes at least 9 months to occur. 3D gait analysis for assessment of patients undergoing anterior cruciate ligament reconstruction surgery can precisely reflect knee biomechanical changes, and assist to explore optimal treatment approach and postoperative rehabilitation methods. PMID- 20017323 TI - [Second-look arthroscopy study after anterior cruciate ligament reconstruction: effect of residual anterior cruciate ligament tissue upon accelerating revascularization of allologous bone-patellar tendon-bone grafting]. AB - OBJECTIVE: To explore via an arthroscopic second-look the effect of preserved residual anterior cruciate ligament (ACL) tissue upon revascularization of allologous bone-patellar tendon-bone (B-PT-B) grafting. METHODS: Twenty-eight patients with allologous B-PT-B ACL reconstruction were recruited. Patients were divided into 2 groups according to the amount of residual ACL tissue. Group A: ACL residual tissue was cleared; Group B: ACL residual tissue was preserved. Patients were advised to undergo arthroscopic second-look at 6 months after ACL reconstruction. During second surgery, ACL photos were taken during 10 minutes after tourniquet was released. Photoshop was used to analyze red chroma of ACL tissue. The percentage area of three degrees of red chroma was calculated and a standard curve plotted. RESULTS: Fourteen cases in group A and 13 cases in group B were examined by second-look arthroscopy. According to Photoshop analysis, ACL graft in group B had better red chroma appearance than group A. Two peaks were observed on grade three red chroma curve. CONCLUSION: Residual ACL tissue can be preserved during allologous B-PB-B reconstruction of ACL At 6 months post reconstruction, the revascularization of graft with preserved residual ACL tissue was much better than the other grafts. Small vessels were found on surface of all ACL grafts at 6 months post-reconstruction. PMID- 20017324 TI - [RIGIDfix tibial and femur cross pin system used for hamstring grafted anterior cruciate ligament reconstruction]. AB - OBJECTIVE: To evaluate the curative effect of RIGIDfix tibial and femur cross pin system used for hamstring grafted reconstruction of anterior cruciate ligament (ACL) in arthroscopy. METHODS: Thirty two cases with ACL ruptures were reconstructed arthroscopically with hamstring grafts and the grafts fixed with RIGIDfix cross-pin on both femoral and tibial sides. They included 18 males and 14 females with a mean age of 28 (20-45) years old; the mean time from injury to operation was 3.6 (2.5-6) months. And each knee was checked by MRI and X-ray preoperatively. Through arthroscopy, we found 19 ACL ruptured from femoral attachment point, 13 from tibial point, 3 cases combined with medial meniscus injury and 4 cases with articular cartilage injury of medial femoral condyle. After semitendinosus or gracilis harvesting, the hamstring grafts were pre tensioned and woven, the diameter of 4 or 5 strands grafts was 7-8 mm. To position and drill tibial tunnel on ACL stump of tibial crest, and to drill femoral tunnel at 10:00 to 11 o'clock of femoral intercondylar fossa transtibial with knee flexed to 90 degree, the depth of femoral tunnel was 30 mm. The RIGIDfix guide was inserted through tibial tunnel up into the femoral tunnel to drill the sleeve and interlocking Trocar across the lateral femur and keep the two sleeves fixed to the lateral femur. Insert RIGIDfix guide into the tibial tunnel, keeping the top of guide 2-3 mm beneath the endostoma of tibial tunnel, drill the sleeve and interlocking Trocar across the lateral tibia and keep the two sleeve fixed to the lateral tibia. The graft was pulled into the tunnel (the 30 mm mark on the graft should be at the edge of femoral hole) to insert the RIGIDfix cross pins from femoral tunnel to tibial tunnel, while inserting the second, third, fourth cross pin, the graft should be kept under tension. Then a knot was tied through tibial bony bridge using the Enthibond thread switched to the end of tendon grafts. We finally observed the tension of tendon grafts and the impingement of fossa intercondylic under arthroscopy. RESULTS: With a 16 month follow-up evaluation, all of the patients' injured knees were stable and the average Lysholm knee score increased from 62.5 to 94.5. Rulermetr device values were less than 2 mm of sagittal displacement in 28 patients and 4 mm in 4. Postoperative Lachman was negative in 30 patients and weakly positive in 2. According to the IKDC scores, 30 patients reported normal function, 2 reported nearly normal function and none reported abnormal or severely abnormal function. CONCLUSION: The grafting method of fixing both femur and tibia sides with absorbable cross pins is feasible. In this way, the graft is stabilized to allow for reconstruction. A surgeon should refrain from dissecting the tendon and enlarging the tunnel so as to promote the healing of tendon and bone. PMID- 20017325 TI - [Effect of maternal antepartum psychological therapy upon early infant temperament]. AB - OBJECTIVE: To find out whether there was any change in early infant temperament after mothers had received group psychological therapy on depression and anxiety during pregnancy period. METHODS: A total of 800 subjects meeting the inclusion criteria, without the exclusion criteria and willing to sign the informed consent were recruited randomly from Shanghai International Peace Maternity & Child Health Hospital in their l6th-20th weeks of pregnancy. They were randomized into the therapy group and the control group by the doll randomization table. Women in the therapy group would have a group psychological therapy for 6 times, 1.5 hours each time, while the control group not. The group psychological therapy included therapist introduction and participatory discussion. The therapy concerned the antepartum and postpartum depression, the risk factors concerned with antepartum and postpartum depression, antepartum and postpartum anxiety, psychological defense theory, reflex training and spouse lesson. Mothers reported their babies' infant temperament by filling the early infant temperament questionnaire (EITQ) within 3 months post-delivery. RESULTS: The percent of easy infant temperament type was 87.9% in the psychological therapy group and 81.7% in the hard group. And there was significant difference between two groups (chi2 = 4.530, upsilon=1, P = 0.033). And there were significant differences in the dimensions of approach withdrawal and threshold of responsiveness in infant temperament between two groups. CONCLUSION: The maternal antepartum psychological therapy can increase the ratio of easy infant temperament. And it has effects upon the temperament dimensions of approach-withdrawal and threshold of responsiveness. PMID- 20017327 TI - [CT guided radio frequency ablation as a treatment of pelvis tumor]. AB - OBJECTIVE: To explore the clinical value of CT guided radio frequency ablation in the treatment of pelvis tumor. METHODS: 11 males and 8 females ranging from 25 to 49 years old (average 37) were enrolled in this study, all the 19 patients were suffering from pelvis tumor including giant cell tumor of bone (2/19), Ewing's sarcoma (2/19), malignant fibrous histiocytoma (3/19) and metastatic diseases (12/19). The average diameter of the lesions were 6.2 cm. The predominant clinical symptom of the patients was pain in the local region. All the patients were treated by CT guided radio frequency ablation. Single pole or multiple poles radio frequency ablation device was used according to the characteristics of the tumor tissue. Time of therapy was 5-15 min. Superposition was performed if necessary. RESULTS: All the patients received PET-CT scan or enhanced CT scan 1 month after the treatment, CR, PR was seen in 10 and 7 cases respectively. Total pain relief was seen in 10 patients, partial pain relief was seen in 8 patients, and gently pain relief was seen in 1 patient. CONCLUSION: CT guided CT guided radio frequency ablation is an effective methods in the treatment of pelvis tumor. PMID- 20017326 TI - [Early diagnosis of endometrial disorder in women with postmenopausal bleeding by three-dimensional transvaginal sonography and hysterosonography]. AB - OBJECTIVE: To evaluate the value of three-dimensional transvaginal sonography and hysterosonography in the diagnosis of endometrial lesion in women with postmenopausal bleeding. METHODS: A total of 74 patients with postmenopausal bleeding were prospectively studied by two- and three-dimensional transvaginal sonography. Endometrial thickness and volume were measured by 2D and 3D ultrasound respectively. Endometrial thickness was found by hysterosonography at greater than 5 mm in 32 cases. Changes of endometrium and compliance of muscularis uteri were observed. The results were correlated with histopathological findings. RESULTS: When 10 ml of endometrial volume was set as the cut-off point of endometrial abnormalities in postmenopausal women with bleeding, the sensitivity and false positive rate were 95.4%, 21.2% whereas 100%, 18.2% were obtained respectively when it was used to predict endometrial malignancy and premalignancy. The accurate rate of 2D-HSG diagnosing endometrial disorders and endometrial malignancy and pre-malignancy was 96. 8% and 100% respectively. Changes of endometrium and compliance of muscularis uteri showed a significant difference between the groups of benign and malignancy after the injection of fluid into uterine cavity (P < 0.05). CONCLUSION: 3D-TVS and 2D-HSG are reliable methods in screening endometrial lesions. 2D-HSG should be regarded as an important tool in diagnosing and differentiating endometrial lesions especially in early stages of this malignant disease. Changes of endometrium and compliance of muscularis uteri may help to differentiate benign disorders from endometrial cancer. PMID- 20017328 TI - [Transcallosal-interforniceal approach treatment of hypothalamic astrocytoma in children]. AB - OBJECTIVE: To explore the clinical features, diagnosis and efficiency of transcallosal-interforniceal approach treatment of hypothalamic astrocytoma in children. METHODS: A total of 42 pediatric cases of hypothalamic astrocytoma were operated by transcallosal-interforniceal approach. All cases received post operative radiotherapy and follow-up visits. The clinical and imaging data were analyzed retrospectively. RESULTS: High ICP and hypothalamic lesions were the main clinical manifestations. All patients suffering from hydrocephalus were operated by transcallosal-interforniceal approach. Seventeen cases received subtotal resection and 25 in great part. None died from operation. All cases received post-operative radiotherapy. Twenty-nine cases were followed up for 1 to 7 years. Among which, 25 led a normal life and 4 stayed in a fair condition. CONCLUSIONS: Hypothalamic astrocytoma in children have some characteristic in clinical manifestations and neuroradiological examinations. Transcallosal interforniceal approach is an effective method of treating hypothalamic astrocytoma in children. Total removal is not imperative. And radiotherapy should be performed. PMID- 20017329 TI - [Characteristics of abnormal glucose tolerance in patients with polycystic ovary syndrome]. AB - OBJECTIVE: To analyze the characteristics of abnormal glucose tolerance in patients with polycystic ovary syndrome (PCOS). METHODS: A 75 g oral glucose tolerance test (OGTT) with plasma glucose and serum insulin (INS) levels was performed in 257 PCOS patients. The states of glucose tolerance were classified by fasting plasma glucose (FPG) and plasma glucose at 120 minutes according to the Word Health Organization (WHO) criterion. RESULTS: According to the WHO criteria, 69.3% were classified as normal glucose tolerance (NGT) and 30.7% as with abnormal glucose metabolism, with 26.1% of them presenting impaired glucose tolerance (IGT) and 4.7% diabetes mellitus (DM). Age, BMI, fasting glucose or 2 h glucose concentration, fasting insulin or 2 h glucose concentration and homeostasis model assessment-insulin resistance (HOMA-IR) increased gradually among NGT, IGT and 2DM. QUICKI and the ratio of fasting glucose/ fasting insulin decreased gradually. The prevalence of abnormal glucose tolerance was significantly higher after 30 years old. There was significant correlation between abnormal glucose tolerance and age or BMI. A positive correlation was found between BMI and fasting glucose or 2-h glucose concentration. CONCLUSION: Abnormal glucose tolerance was correlated with age and BMI in PCOS patients. Insulin resistance is the mechanistic basis of abnormal glucose tolerance. Thus PCOS patients should be screened by glucose tolerance at an early time and a regular interval. PMID- 20017330 TI - [Different treatment modalities of patients with stage III lung cancer]. AB - OBJECTIVE: In order to improve the survival-rates and study the long-term therapeutic effect of stage III lung cancer patients with different treatment modalities. METHODS: From 1995 to 2004, 1921 patients of stage III lung cancer treated at our hospital were retrospectively analyzed to compare the long-term therapeutic effect of different treatment modalities. There were 606 cases in the combined therapy group (mainly surgical with combined treatment of chemotherapy & radiotherapy), 317 cases in the surgery alone group and 998 cases in the non surgical group. The 1, 3, 5-year survival rates of these three groups were 66.0% (400/606) vs. 62.7% (199/317) vs. 51.2% (511/998), 32.34% (196/606) vs. 21.45% (68/317) vs. 8.15% (81/998), 19.31% (117/606) vs. 13.2% (42/317) vs. 4.2% (42/998) respectively. In comparison, 1, 3, 5-year survival rates of the combined therapy group was higher than the surgery alone and non-surgical groups; Significant differences in 1-year survival rates were found between the combined therapy group and the surgery alone group or the non-surgical group (P < 0.01) and significant differences in 3, 5-year survival rates among these three groups. CONCLUSION: The combined therapy of radical resection with chemotherapy and radiotherapy is the best treatment modality for stage III lung cancer. PMID- 20017331 TI - [A multicenter randomized controlled study of temozolomide in 97 patients with malignant brain glioma]. AB - OBJECTIVE: To investigate the efficacy and safety of temozolomide (TMZ) and lomustine (CCNU) in malignant brain gliomas. METHODS: In this multicenter, randomized, double-blind, positive-controlled study, patients with malignant glioma of a histological grade III/IV were randomly divided into two groups. In experiment group, TMZ capsule 150 mg/m2 was given orally on days 1 through 5 and CCNU placebo 130 mg/m2 while on day 1. In control group, CCNU capsule 130 mg/m2 was taken orally on day 1 and TMZ placebo 150 mg/m2 while on days 1 through 5. In both groups drugs were given on the first 5 days of 28 day therapeutic course. Every patient was administered at least 3 cycles continuously. RESULTS: 97 patients were enrolled in the clinical trial, 86 cases of which were evaluable for efficacy. The basic conditions of patients in both groups were comparable. After 12 weeks, the response rates in TMZ and CCNU group were 35.71% and 9.09% (P < 0.01) respectively, and the clinical benefit rates in TMZ and CCNU group were 90.48% and 75.00% (P < 0.05) respectively. No obvious improvement was showed in quality of life and neural symptoms in both groups. The common side effects of TMZ were nausea and vomiting with grade I/II. CONCLUSION: TMZ capsule can produce higher efficacy than CCNU in the treatment for refractory malignant brain glioma patients with an acceptable safety profile , indicating that TMZ could be an ideal chemotherapy selection for malignant brain glioma. PMID- 20017332 TI - [Experimental study of implantation of neurotropin-3 modified olfactory ensheathing cells in experimental allergic encephalomyelitis]. AB - OBJECTIVE: To investigate the therapeutic effect of neurotrophin-3 (NT-3) modified olfactory ensheathing cell (OEC) upon experimental allergic encephalomyelitis (EAE). METHODS: OEC-NT-3 gene engineering cell, constructed by neurotrophin-3 transinfecting GEC inducted by retrovirus, was transplanted into lateral ventricle. The migration and distribution were observed and compared with control group and OEC transplantation group. Then myelin repairing and axon regeneration were evaluated from conical somatosensory evoked potential (CSEP), function score and ultrastructural morphology. RESULTS: (1) OEC-NT-3 could survive, migrate within axons and spread diffusely away from the focus at Day 28 post-transplantation; (2) as compared with other two groups, more nerve fibers, better myelin repair and more distinct myelin structure were observed in the transgene group; (3) as compared with other two groups, the latent time was obviously shortened and the amplitude higher in the transplantation group (P < 0.05); (4) the transcription level of NT-3mRNA in the transgene group was significantly higher than the GEC group and the contrast group (212.32 +/- 16.14) x 10(-2) vs. (1.98 +/- 0.19) x 10(-2), (1.23 +/- 0.13) x 10(-2) (P < 0.01). CONCLUSION: OEC-NT-3 cell expresses NT-3 stably and effectively in EAE. It may contribute to the repairing of myelin and the regeneration of axon. PMID- 20017333 TI - [Gene expression of OAZ pathway in the patients of systemic lupus erythematosus]. AB - OBJECTIVE: To explore the role of Olf/EBF associated zinc finger protein (OAZ) pathway in the RNA expression level in patients with systemic lupus erythematosus (SLE). METHODS: The expression levels of OAZ, BMP6, BMP4, Id3, Smad6, EHZF genes were valuated in bone marrow progenitor cells of 5 SLE patients and 5 normal subjects and replicated in peripheral blood cells of 30 SLE patients and 20 normal individuals by using real-time quantitative PCR technique. Relationships of the expression levels of OAZ, BMP6, BMP4, Id3 mRNA with disease activity and other clinical indices were analyzed. RESULTS: The expression levels of OAZ and Id3 mRNA (deltaCt) in the bone marrow (10.6 +/- 0.5, 5.8 +/- 3.2) and peripheral blood (14.1 +/- 2.7, 7.5 +/- 1.8) of SLE patients significantly increased than those observed in normal controls (16.5 +/- 0.9, 10.4 +/- 2.6, 16.1 +/- 2.2, 9.5 +/- 1.7), which was found to negatively correlate with SLEDAI score, renal lesion index , titers of anti-dsDNA and anti-RNA antibodies, but positively correlate with serum complement C3. Expression levels of BMP4 and BMP6 were differentially expressed in peripheral blood cells but not bone marrow progenitor cells. CONCLUSIONS: OAZ pathway is involved in the pathogenesis of SLE. Further investigation is required for the understanding of the function of these abnormally expressed genes. PMID- 20017334 TI - [Effects of radiotherapy upon progression of crescentic glomerulonephritis in rats]. AB - OBJECTIVE: To study the effects of radiotherapy upon progression of crescentic glomerulonephritis in rats. METHODS: Male SD rats were divided into three groups: (1) control (n=12), sham-operation; (2) crescentic glomerulonephritis (n=23), intravenously inject with nephrotoxic serum (NTS); (3) radiotherapy (n=55), a single low-dose irradiation of 0.5 Gy X-ray to both kidneys at Days 6, 13, 20 and 27 after NTS injection, and sacrificed at different time points among control and crescentic glomerulonephritis rats. Radiotherapy rats have received local kidney irradiation at Days 6, 13, 20 and 27 after bolus NTS injection and would be referred to as NTS7dRa1d, NTS14dRa1d, NTS21dRa1d and NTS28dRa1d, respectively. RESULTS: For NTS7dRa1d and NTS14dRa1d rats of radiotherapy, the levels of serum creatinine, glomerular hypercellularity, crescents and global sclerosis were significantly lower at Days 8 (P < 0.05), 15, and 22 post-irradiation as compared with group of crescentic glomerulonephritis of similar time intervals (P < 0.01). The extent of tubulointerstitial damage was also reduced, and radiotherapy associated histological improvements were accompanied by reduced macrophage infiltration in glomeruli and interstitium. The numbers of PCNA- and ED1-positive cells were reduced in the kidneys at Day 1 postirradiation in NTS7dRa1d and NTS14dRa1d rats as compared with group of crescentic glomerulonephritis at similar time intervals (P < 0.05). A larger number of TUNEL-positive cells were noted at Day 1 postirradiation in rats irradiated at Days 6 & 13 after NTS injection as compared with group of crescentic glomerulonephritis at similar time intervals (P < 0.05). With regards to immunostaining for macrophages ED1 and TUNEL, serial sections of irradiated nephritic kidney showed that fewer ED1 positive macrophages were stained for TUNEL. As evaluated expression of active caspases 3 & 7 was noted in irradiated kidneys as compared with the corresponding group of crescentic glomerulonephritis at similar time intervals. Western blot analysis showed marked increase in the expression of active caspase 3 & 7 in irradiated kidneys as compared with NTS injection only the expression of a marked increase in the expression of p53 protein, closely related to radiation-induced apoptosis, was also observed in irradiated kidneys as compared with NTS injection only. CONCLUSION: Radiotherapy inhibits the progression of experimental crescentic glomerulonephritis through inducing apoptosis by a p53-dependent pathway. PMID- 20017335 TI - [Effect of propofol upon ammonia-induced neocortical astrocyte swelling and aquaporin-4 expression]. AB - OBJECTIVE: To investigate the effect of propofol upon ammonia-induced neocortical astrocyte swelling and aquaporin-4 expression in rat astrocyte. METHODS: Methods Astrocytes were isolated from newborn Sprague Dawley rats. After a 3-week culture, cell immunofluorescence was employed to label glial fibrillary acidic protein, the specific protein of astrocyte. Astrocytes were cultured with NH4Cl (5 mmol/L) for 6, 12, 24 and 48h. Astrocytes were pretreated by propofol and DMSO respectively for 30 min and then exposed to NH4Cl for 24h. The expression of AQP4 was detected by the Western blot; cell morphology assessed by light microscopy and cell viability measured by MTF reduction assay. RESULTS: The expression of AQP4 was elevated after a 12h exposure to ammonia, lasted to 48h and peaked at 24h. Astrocytes were found significantly swelling in the NH4Cl-24h and DMSO pretreated groups as compared with the propofol pretreated group. The over expression of aquaporin-4 was attenuated by propofol pretreatment and the cell viability of astrocytes in the propofol pretreated group was higher than that in the NH4Cl-24h group (P < 0.05). CONCLUSION: Pretreatment of astrocytes with propofol can inhibit the over-expression of AQP4, relieve cellular swelling and reduce the ammonia-induced decline in cell viability of astrocytes. PMID- 20017336 TI - [Electrophysiological characteristics of neurons in rat substantia nigra by visual patch clamp technique]. AB - OBJECTIVE: To establish the brain slices of substantia nigra and striatum in rats and study the properties and functional significance of substantia nigra neurons. METHODS: The technique of infrared differential interference contrast (IR-DIC) and CCD-Camera system with visual patch clamp whole-cell recording were combined. One hundred healthy substantia nigra neurons were located and the measurements of active potential and Ih current recorded and analyzed. RESULTS: Visual patch clamp technique could be used to make a direct localization of substantia nigra and to identify the active neurons. Spontaneous active potential was shown in 94/100 neurons. Among them, 85 (Group A) had a frequency less than 10 Hz and 9 more than 10Hz (Group B). The means were (5.0 +/- 2.7) Hz and (14.0 +/- 2.6) Hz. respectively. As compared with the former, the latter had no obvious hyperpolarization during active potential period. Apparent hyperpolarizing current could be induced in 92.9% (79/85) of Group A neurons (-120 mV, 47.5 pA +/ 5.8 pA). They were classified as dopaminergic neurons. However, the Group B neurons had no or little Ih current and were classified as GABA neurons (P < 0.001). CONCLUSION: The types of nigra neurons can be distinguished by observing the fundamental electrical characteristic of dopaminergic neurons in nigrostriatal brain slices under visual inspections. And it may provide rationales for an in-depth knowledge of the occurrence and transmission of signal pathways between substantia nigra and PMID- 20017337 TI - Achieving excellence through employee commitment. PMID- 20017338 TI - Nurturing residents' spirits. PMID- 20017339 TI - HIT management made easy. PMID- 20017340 TI - Nutrition under a microscope. PMID- 20017341 TI - Single fuel source offers savings. PMID- 20017342 TI - [Non-medicamentous methods for the treatment of metabolic syndrome]. AB - Non-medicamenous methods for the treatment of metabolic syndrome (MS) are discussed. Diagnosis of MS is a priority problem of modern medicine and healthcare because of its important role in pathogenesis of type 2 diabetes, essential hypertension, and atherosclerosis complicated by coronary heart disease, i.e. major causes of mortality worldwide. Non-medicamentous treatment permits to diminish the total amount of pharmaceuticals prescribed to the patients and to improve the overall result of drug therapy. These methods include remedial gymnastics, physiotherapy, reflexotherapy, balneological treatment, and psychotherapy. Different modalities of non-medicamentous therapy are considered with special reference to the methods for lowering body weight as an earliest and most dangerous manifestation of MS. Indications and contraindications to the use of concrete therapeutic modalities are described. PMID- 20017343 TI - [Syndrome of chronic duodenal obstruction: modern view of the problem]. AB - This paper reviews data on the prevalence and clinical significance of syndrome of chronic duodenal obstruction (CDO), duodenal anatomy and physiology. The definition of CDO is proposed along with classification of its organic and Functional forms, their characteristics, and clinical manifestations at the stages of compensation, sub- and decompensation. Instrumental and laboratory diagnostic methods are discussed, their effectiveness and informative value are assessed. Comprehensive review of the principles of treatment of functional and organic forms of CDO (nutritional treatment, physiotherapy, drug therapy, remedial gymnastics, surgery) is presented. PMID- 20017344 TI - [Preventive strategy for atrial fibrillation in arterial hypertension]. AB - The aim of the study was to evaluate effectiveness and safety of angiotensin II receptor blockers and inhibitors of angiotensin-converting enzyme (ECE) for protective therapy following medicamentous cardioversion with propafen at a loading dose of 600 mg in patients with paroxysmal atrial fibrillation and arterial hypertension. 101 patients were divided in 2 groups. Group 1 included 75 patients who received ACE inhibitor lisinopril (10 mg BID) after recovery of sinus rhythm by propanorm. 26 patients of group 2 were treated with angiotensin II receptor blocker candesartan (8 mg daily). Combined treatment with angiotensin II receptor blocker and propafenone leads to cardioversion faster than therapy with ACR inhibitor. It is concluded that alternative approach to the maintenance of sinus rhythm using angiotensin II receptor blockers has advantages over traditional anti-arrhythmic therapy; it is well tolerated by the patients and produced no serious side effects. PMID- 20017345 TI - [Arterial hypertension and type 2 diabetes mellitus: clinical evaluation of hemodynamic characteristics, possibility of correction]. AB - The study was designed to evaluate effect of enalapril and telmisartan on hemodynamic characteristics and diastolic function (DF) of left ventricle (LV) in patients with type 2 diabetes and arterial hypertension (AH). It included 64 patients aged 54.3 +/- 5.2 years. Those in group 1 (n = 31) were given enalapril (enap), patients of group 2 (n = 33) were treated with telmisartan (micardis). Examination included 24 hour AP monitoring, Holter ECG monitoring, and echocardiography. Compensation of metabolic disorders was evaluated from fasting and postprandial blood glucose and HbAc1 levels. Impaired LV DF was the main feature of affected myocardium in patients with DM2 and elevated AP in the absence of contractility disturbance. Enalapril therapy ensured the desired level of systolic and diastolic AP in 77 and 64.5% of the patients respectively in association with a decreased number of non-dippers and night-peakers in 45.4% of the observations in the absence of changes in HbAc1 level and LV DF. Treatment with telmisartan ensured within 24 weeks efficacious control of systolic AP and normalization of its daily profile in 87.5% patients with pathological circadian rhythm, besides improvement of carbohydrate metabolism and LV DF. PMID- 20017346 TI - [Drug therapy of diastolic cardiac insufficiency in patients with type 2 diabetes and concomitant essential arterial hypertension]. AB - Therapeutic efficiency of spirapryl and its combination with carvedilol in the treatment of diastolic cardiac insufficiency in patients with type 2 diabetes and essential arterial hypertension (AH). Criteria for inclusion in the study of 86 patients (20 men and 66 women) were diastolic cardiac insufficiency diagnosed as recommended by the Working Group of European Society of Cardiologists (2002). The patients were divided into 2 groups receiving either spirapryl (n = 42) or spirapryl with carvedilol (alpha, beta-adenoblocker). The patients were examined before and 24 weeks after therapy for the evaluation of their clinical condition, tolerance of physical activity, transmitral and transcuspidal diastolic flows (ECG and Doppler-ECG). All therapeutic modalities significantly improved the patients" conditions, diastolic function, and ECG/ECG characteristics, but the best results were obtained with combined therapy that had beneficial effect on transmitral and transcuspidal diastolic flows (P < 0.01) and tolerance of physical exercises (P < 0.05). It is concluded that combined treatment with spirapryl and its combination with carvedilol is more efficient for the management of diastolic cardiac insufficiency than carvedilol monotherapy. PMID- 20017347 TI - [Diastolic dysfunction of left ventricle in psoriatic patients with concomitant opisthorchiasis before and after antihelminthic therapy]. AB - The aim of the study was to evaluate left ventricular diastolic function in psoriatic patients with chronic opisthrochosis (CO) before and after antihelminthic therapy. It included 75 patients with psoriasis and CO, 50 with psoriasis alone, 50 with CO, and 20 healthy subjects. All patients underwent standard echocardiographic examination in the M-, B-, and Doppler mode. The study revealed impaired active relaxation of the left ventricle manifest as increased isovolumic relaxation time more pronounced in patients with concurrent CO and psoriasis than in those with psoriasis alone. Antihelminthic therapy did not cause deterioration of the diastolic function in these patients and should be recommended in case of combined pathology. PMID- 20017348 TI - [Scintigraphic evaluation of cerebral circulation in prognosis of cerebrovascular complications of coronary artery bypass grafting]. AB - The aim of the work was to study the possibility of using single-photon emission computed tomography (OECT) of brain using 99m Tc-hexamethyl propylene aminoxime for the evaluation of long-term prognosis of the development of neurocognitive deficit after coronary artery bypass grafting (CBG) and elucidation of relationship between variations of cognitive function and regional cerebral circulation. Surgical myocardial revascularization under artificial circulation was performed in 22 patients with CHD. All of them were examined for the assessment of neurologic and neuropsychological status, and OECT data before, 14 days and 6 months after CBg. Early cognitive deficit developed in 96% of the patients and persisted for 6 months in 55%. Cognitive dysfunction was manifest as post-operative impairment of short- and long-term memory, and general efficiency of remembering. Visual memory and psychomotor speed also deteriorated. Cerebral circulation decreased in frontal and parietal lobes of the right hemisphere. Deterioration of long-term memory 6 months postoperatively correlated with the early circulation decrease of in the right and left temporal lobes. PMID- 20017349 TI - [Radionuclide renoscintigraphy for the evaluation of nephroprotective action of acetylcysteine]. AB - Radionuclide renoscintigraphy was used to evaluate efficiency of nephroprotection by N- acetylcholine in 36 patients subjected to coronaroangiography (CAG). The patients were randomly allocated to 2 groups, one undergoing CAG without nephroprotection, the other CAG in combination with N-acetylcholine nephroprotector. Dynamic radionuclide renoscintigraphy with 99m Tc-dtpa was performed before and 2-3 days after CAG; the results were used to estimate filtration and evacuation efficiency of the kidneys. Patients of group 1 showed negative dynamics of the renal function following CAG including decreased mean glomerular filtration rate and slowed down blood clearance. Mean half-time of 99mTc elimination from the pelvicalyceal system and parenchyma of the left kidney as well as its cortical retention index in bot kidneys increased. The filtration activity of the kidneys did not significantly change after CAF in patients of group 2; it was slightly impaired only in 5 (28%) of them. More patients suffered deterioration of 99mTc elimination but on the whole changes in the evacuation activity of both parenchyma and pelvicalyceal system were statistically insignificant. It is concluded that acetylcholine has nephroprotective action in patients subjected to radionuclide renoscintigraphy. PMID- 20017350 TI - [Peculiarities of the treatment of obstructive pulmonary disease in patients with combined pathology]. AB - Effects of combined drug therapy were evaluated in 97 patients during a year Group 1 comprised 32 patients with chronic obstructive pulmonary disease (COPD), group 2 (n = 34) included patients with COPD and angina of effort, group 3 (n = 31) patients with CHD. Broncholytic therapy was prescribed in compliance with GOLD (2006) recommendations. Patients in groups 2 and 3 continued to receive antianginal therapy started before the onset of the study. Combined therapy included ACE inhibitor enalapril. Positive effect of inpatient treatment of COPD on lung function was transient and subsided with time. Specifically, forced expiratory volume in the 1st second and forced pulmonary vital capacity decreased below the acceptable physiological bounds. BODE index tended to drop too because exercise tolerance increased while dyspnea index by MMRC decreased despite impaired FEV1 and tendency toward a fall in BMI. Echocardiography revealed diminished size of both ventricles and improved left ventricular systolic function. However, heart remodeling progressed with time. Adequate therapy resulted in the reduction of CHD functional class and produced positive antianginal, antihypertensive, and antiarrhythmic effects. The treatment was well tolerated by the patients. It is concluded that combined therapy of COPD including enalapril improves respiratory symptoms and decreases manifestations of concomitant pathology. PMID- 20017351 TI - [Diagnostic criteria for different clinical variants of diverticular disease]. AB - New diagnostic criteria for different variants of diverticular disease (DD) are considered based on the examination of 110 patients. Patients of group 1 (n = 77) showed no signs of diverticulitis in contrast to group 2 (n = 33). Two control groups comprised 38 patients with chronic hypomotor colitis and 25 practically healthy subjects respectively. Dynamic observations included clinical and endoscopic examination supplemented by morphological and immunohistochemical Studies. It was shown that DD developed in association with undifferentiated dysplasia of connective tissue, increased number of P substance-inducing colonocytes, and lowered density of vasointestinal peptide-reactive cells. Diagnostically significant criteria in patients with diverticulitis were grade II III intestinal dysbiosis and increased number of mast cells in rectosigmoid nucosa. PMID- 20017352 TI - [Splenorectal blood flow in chronic viral hepatitis B and C]. AB - The aim of the study was to compare results of dopplerography characterizing hepatic and splenic blood flow at different stages of chronic viral hepatitis B and C and correlate them with histological findings and sclerosis. The study included 79 patients of whom 45 had chronic hepatitis B (CHB) and 34 hepatitis C (CHC). The most sensitive dopplerographic characteristics proved to be congestion index, liver vascular index, and hepatic hypertension index that started to change significantly at minimal activity of hepatitis and greatly deteriorated after development of liver cirrhosis. In both cases venous blood flow was affected more seriously than arterial one. Hepatic hyperperfusion progressed faster in CHC than in CHB. Dopplerographic characteristics correlated with histologic activity index, its components, and sclerosis index. Hence, the possibility of using dopplerographic studies for the evaluation of hepatic and splenic vasculature and indirect assessment of morphological changes in the liver. PMID- 20017353 TI - [Psychosomatic relationships in patients with primary fibromyalgia syndrome]. AB - This clinical and psychological study involved 100 patients aged 24-51 years with primary fibromyalgia syndrome (PFS). The patents exhibited clinical manifestations of neurotic disorders, low level of subjective control, dysadaptive attitude toward the disease, and an intense spectrum of protective psychological mechanisms. The main clinical manifestations of PFS correlated with psychological status of the patients. It is concluded that objective evaluation of clinical conditions of PFS patients requires integrated psychosomatic assessment of their health status. PMID- 20017354 TI - [Neurologic manifestations of HIV infection]. AB - The sole universally recognized diagnostic criterion for the affection of central nervous system by immunodeficiency virus is AIDS-dementia complex. Its main pathognomonic feature is progressive intellect failure although most patients do not experience intellect deficit. Instead, they present with other disorders in the central nervous system. This article focuses on their early diagnosis. PMID- 20017355 TI - [Use of prestarium (perindopril) in patients with polymorbidity syndrome (AH, DM, nephropathy)]. AB - The study included 20 patients aged 61-86 (mean 73.1 +/- 3.1) years with stage II hypertensive disease, type 2 diabetes and nephropathy with signs of grade I chronic renal insufficiency (CRI). The major parameters measured were AP, HR, laboratory characteristics of carbohydrate metabolism and renal function. Prestarium given at a dose of 4 mg BID had not only antihypertensive but also negative chronotropic effect, decreased energy consumption by myocardium, and normalized daily AP profile (24 hr AP monitoring). Adequate control of diabetes ensured close-to-normal carbohydrate metabolism. Prestarium therapy did not cause further aggravation of renal insufficiency in patients with grade I CRI, nor did it alter creatinine and urea levels. PMID- 20017356 TI - [Comparative analysis of effectiveness and safety of diclofenac and nimesulide in patients with early rheumatoid arthritis]. AB - The aim of the work was to evaluate effectiveness and safety of diclofenac and nimesulide in patients with early RA. The open clinical study included 80 outpatients (mean age 49.0 +/- 11.1 years) within the first year after development RA (mean duration of the disease 4.9 +/- 3.1 months prior to the onset of basic therapy). The patients were divided into 2 groups of 40 persons each depending on the type of non-steroidal anti-inflammatory agents they received. Patients in group 1 were treated with 100 mg of diclofenac daily and in group 2 with 100 mg BID of generic nimesulide preparation (nais, Dr. Reddy Co.). Duration of therapy was 30 days in both cases. The two drugs were roughly identical in terms of therapeutic effect although nimesulide produced fewer side effects. Subjective complaints of gastrointestinal problems were recorded in 11 (27.5%) patients f group 1 and in 8 (20%) of group 2. It is concluded that timely prescription of proton pump inhibitors permits to avoid premature withdrawal of non-steroidal anti-inflammatory agents at patient discretion. PMID- 20017357 TI - [Use of three-dimensional ultrasound visualization in differential diagnosis of pancreatic head diseases]. AB - Retrospective analysis of 3D-reconstruction of ultrasound images is discussed with reference to the diagnosis of pancreatic disorders, such as pseudotumorous pancreatitis and pancreatic head neoplasm, based on the data extracted from case records collected in the Central Clinical Hospital No 1, OJSC RZhD, for the period of 2004-2008. In addition, comparative analysis is presented of different beam diagnostics methods for the examination of hepatobiliary system (classical ultrasound, 3D-reconstruction of ultrasound images, CT, endoscopic techniques, e.g. retrograde cholangiopancreatography). Control group comprised 20 patients without signs of gastrointestinal pathology, group 2 included 24 patients with pseudotumorous pancreatitis, group 3 54 patients with pancreatic head neoplasm (up to 3 cm). It was shown that 3D-images facilitate detection of pathologic processes in the pancreas. Minimal slice thickness permits to visualize small objects, segmentation of irregularly shaped tumorous foci and their volume. This method gives better idea of the relationship between the tumour and other tissues and vessels than other diagnostic tools and thereby makes easier the choice of treatment strategy and prognosis of its outcome. Also, it improves the quality of information about arterial and venous circulation and permits to differentiate between benign and malignant tumours. PMID- 20017359 TI - [Genotyping of the Bacillus anthracis vaccine strains using the multiloci VNTR analysis]. AB - The genotyping variety of 5 known anthracis vaccine strains using 18 variable loci of the chromosomal localization taken from a microbe culture collection of 48 Research Institute of Ministry of Defense was revealed in the research. The stability of the VNTR-loci was shown to be inherited from the B. anthracis strains with common origin and an opportunity of their gene-identification application. The gene profile of each analyzed vaccine strain using every 18 polymorphic loci was determined and the amplification products were sequenced. The variation of electrophoretic mobility of the amplifiers was found to be caused by the presence of the replication elements with various numbers of copies in their structure. PMID- 20017358 TI - [Poorly differentiated stomach adenocarcinoma combined with jejunal stromal tumour. Clinical study]. AB - Gastrointestinal stromal tumours (GIST) first described in 1983 are the commonest mesenchymal neoplsams of the digestive tract. The main pathogenetic mechanism is hypersecretion of tyrosin kinase receptor c-kit (CD-117) by GIST cells and its hyperactivation. A rare clinical case of GIST measuring 1.2 CM with concomitant stomach cancer (poorly differentiated carcinoma) is reported. The tumours were discovered intraoperatively. To our knowledge, data on such cases are practically lacking in the literature. PMID- 20017360 TI - [Antibacterial effects of silver ions: effect on gram-negative bacteria growth and biofilm formation]. AB - Minimal inhibiting AgNO3 concentration (MICs) in the gram-negative bacteria Escherichia coli K12, Serratia proteamaculans 94, and Serratia liquefaciens MG1 were found to be on the average within the range of 0.075-0.3 microg/ml, and for Pseudomonas aeruginosa PAO1 and P. chlororaphis 449, 0.15-0.3 microg/ml. Biofilm formation in Escherichia coli AB1157 and S. Proteamaculans 94 was completely inhibited at an AgNO3 concentration of 0.3 microg/ml, and in Pseudomonas aeruginosa PAO1, at 0.6 microg/mlAgNO3. Mutations in E. coli genes encoding for global regulators of gene expression, such as sigma S and sigma N subunits of RNA polymerase, catabolite repression protein CRP, and Lon protease, had no marked effect on the sensitivity of cells to silver. The wild-type E. coli strains and strains deficient in excision repair (uvrA, uvrB), SOS-repair or recombination (recA, lexA, recBC, recF mutants) did not differ in their silver sensitivity. This suggests that the sensitivity of bacteria to silver does not correlate with DNA lesions that could be repaired by these repair and recombination systems. E. coli mutant strains deficient in porins OmpF or OmpC, were 3-4-fold more resistant to silver ions as compared with the wild-type strain. Experiments with pME6863 plasmid harboring the gene of N-acyl-homoserine lactonase AiiA demonstrated that Quorum Sensing regulation (QS) did not participate in the control of S. proteamaculans 94 and P. chlororaphis 449 silver sensitivity. The same conclusion was drawn from the comparison of AgNO3 MICs for the S. liquefaciens wild-type strain and a mutant strain deficient in QS. PMID- 20017361 TI - [Hepatitis C virus genotyping using 5 nuclease real-time PCR and probes with oligodeoxyinosine linkers]. AB - Hepatitis C virus (HCV) infection is a major cause of severe liver disease including liver cirrhosis and hepatocellular carcinoma. Genotyping became fundamental in the clinical assessment of patients with hepatitis C because the genotype of the hepatitis C virus determines the chance of therapeutic response and duration of treatment. We developed a new real-time PCR assay for genotyping of HCV with increased specificity due to a novel approach to dual-labeled probe design using oligodeoxyinosine linkers. The assay allows genotypes 1, 2, 3 to be distinguished and genotypes 4-6 with high specificity to be blocked. The analytical sensitivity (150 IU/ml) can be implemented. Of the 285 clinical samples genotyped using the developed assay, 45% were genotype 1; 6%, genotype 2; 49%, genotype 3. No discordant results with 5-UTR sequencing and commercial genotyping assays were obtained. PMID- 20017362 TI - [Human recombinant interferon-B constructed on the basis of affinity-binding domain technology]. AB - Fusion gene consisting of dextran-binding domain from Leuconostoc mesenteroides subsp. Mesenteroides (DBD) and human recombinant interferon-beta (IFN-beta) incorporated between the nucleotide sequence encoding for the recognition site of human enteropeptidase (DDDDK) was installed and constructed in Escherichia coli. The overproducing strain of the chimeric protein DBD-IFN-beta consisting of the IFN-beta, spacer including 10 GS-repeats, human enteropeptidase recognition site, and dextran-binding domain from Leuconostoc mesenteroides was constructed. Free human recombinant interferon-beta was obtained as a result of treatment of the chimeric protein DBD-IFN-beta immobilized on sephadex G-25 with human enteropeptidase. The ability of free and immobilized protein to protect human cells from viral infection was demonstrated. The developed approach can be used for purification of the recombinant proteins with different biological activity and possible construction of new immunostimulating and antiviral drugs, growth factors, anti-cancer drugs, etc. PMID- 20017364 TI - [Pre-morbid borderline state in teenagers]. AB - There was effectuated a clinical-epidemiological, follow-up research of frequency, phenomenological peculiarities and dynamics of pre-existing disease borderline disorders among teenagers-schoolers. There were defined several factors, eintraechtigening on forming stable neurotic and personal disorders. PMID- 20017363 TI - [Psychological and medical rehabilitation of servicemen in sanatorium-treatment institutes of the Defense Ministry of Russian Federation during the current stage of building of the Armed Forces]. AB - Defense and promotion of health of strong and almost strong persons--is one of priorities tasks of health services, including military medicine. Federal law from the 8.11.2008 No 203-FL has made changes in federal law from 1998 No 76FL "About status of military servicemen". These changes allow, in conditions of existing of indications, effectuating psycho-medical rehabilitation less then 30 days free. To execute this federal law the Defense Ministry issued a law No 385 from 8.05.2008 "About psycho-medical rehabilitation of military servicemen of the Defense Ministry of Russian Federation". The law defines catalogue of categories of military servicemen, who are liable to psycho-medical rehabilitation and order of it's effectuating. The article is devoted to comment this law. There was examined in details the role of sanatorium stage in system of medical rehabilitation, and organization of it's effectuating in conditions of sanatorium. PMID- 20017365 TI - [Traumatic stress and personal characteristics of participants in combat operations]. AB - The article presents an analyze of influence of combat traumatic stress on personal characteristics of combat-operations participant of the Afghan war. The research has shown that there are differences in personal peculiarities by veterans, having different psychological pathology. The structure of these violations is interdependent with effective crossing of consequences of psycho trauma. Pathological reactions (chronic post-traumatic stress disorder) is aligned with decrease of psychological functions, emotional disorders (uneasiness, emotional instability, tension) and personal peculiarities (noncomformity, difficultness with understanding of life, low indexes of moral and willed qualities of I-conception). For effective negotiation of consequences of post-traumatic stress it's necessary to solve problems, aligned with understanding of life (finding of purposes and feeling of control under the life) and change attitude to your self in aspect of moral and willed component. PMID- 20017366 TI - [The role of airport evacuation receivers in the system of treatment-evacuation measures]. AB - There was validated the necessity of deployment of inairdrome evacuation receiver for the purpose of evacuation of ill and wounded persons by evacuation transport during organization of massive evacuation. Also there was presented organizational-staff structure of evacuation receiver, material support and principal scheme of deployment. There was deployed one of variants of evacuation receiver (for 100 ill and wounded persons) during tactic-special training exercises. It was concluded that inairdrome evacuation receiver have to be a mobile unit of medical service, which is able to be transported by air transport in areas of massive evacuation of ill and wounded persons. It's proposed structure is recommended as a module for more powerful evacuation receiver. PMID- 20017367 TI - [Extraperitoneal complications in gun-shot penetrating wounds of the stomach]. AB - The article after a short literature review presents data of analyze of progression of extra-peritoneal visceral pathology by wounded persons with penetrating wound of stomach. There was described frequency of morbidity of extra peritoneal organs in dependence on flow of peritonitis. Were marked differences in frequency of progression of peritonitis in conditions of fragment and ballistic wounds. In course of effectuated research was marked dependence between frequency of lethal outcomes and flow of peritonitis and added diseases of respiratory organs, uropoiesis, cardiovascular system. There was effectuated a comparative analysis of frequency of appearance of pathology of marked systems in dependence on localization of ballistic wound. Also there was shortly described a clinical-laboratorial presentation in conditions of appearance of diseases of extra-peritoneal organs. PMID- 20017368 TI - [Prophylaxis of acute respiratory viral infections in organized communities]. AB - There are presented the data of placebocontrolled research, which was realized for the purpose of analyze of effectiveness and safety of using of agent "Kagocel" for prophylaxis of acute respiratory virus infections among military servicemen of call-up service. Among persons, got kagocel, was marked a low number of ARVI in comparison with persons, got placebo. The highest prophylactic effect was marked for the first 2 month of getting the agent. A mild disease (ARVI) flow was marked among persons, got placebo. It has approved it self in low frequency of complications. There was shown a grand economic benefit of prophylactic using of kogocel course of treatment. PMID- 20017369 TI - [Treatment of multiple carbuncles of the kidneys in military servicemen]. AB - The article presents the analyzes of medical histories of 16 patients, treated from suppurative diseases of burgeon (13 carbunculosis, 1 nephrepostasis, 2 abscesses of paranephric cellulose) from 1997 to 2009. Effectuated examinations and treatment of patients with suppurative diseases of burgeon speak to insufficient educating work of the medical service of a military unit in the sphere of prophylaxis if suppurative and catarrhal diseases, what leads to delayed call for medical aid. The second place by level of negative influence of appearance of supparative diseases of urinary organs is taken by conditions of service in northern climate and not full adaptation of service men to these conditions. Changing of food ration also leads to decreasing of immunity and organism general-defense status in conditions of extreme conditions of military service. The article presents indications to operative and conservative treatment of carbunculosis of burgeon. PMID- 20017370 TI - [Medical-physiological characteristics of combat training of nuclear-power submarine crews]. AB - The article presents an observe of general questions of peculiarities of military professional activity of submarine staff These questions are defining value in ideology of medical supply of submarine troops of NAVY in now-days conditions. The article also presents the statistics of morbidity in long termed sails for last forty years, it's dynamics by different categories of sail staff, on different stages of combat training activity in dependence of perioditation of work cycle of submarine staff The authors have examined modern condition of medical supply of submarines; have presented statistics of quality indexes of health of submarine staff The authors have formed main problems of medical supply of submarines and have proposed ways of their solving on modern stage. PMID- 20017371 TI - [Il'ia Vasil'evich Buial'skii--collector of medical rarities]. AB - The article presents the analyze of activity of academician of Saint-Petersburg Medical-surgical Academy Buyal'sky I.V. in sphere of collecting of memorials of medical science and practice, creation of a specialized museum of models of surgical instruments near a factory, and his activity as a painter-aquarellist. The author has paid a special attention to analyze of Buyal'sky catalogs of 1854 and 1867. of Surgical museum, consisting of things from so-called "Cabinet of professor Buyal'sky". For the first time was marked role of Buyal'sky I.V. in elaborating of principles of systematization, defining and observing of historical heritage of native medicine and industry. PMID- 20017372 TI - [Medical care of wounded persons with maxillofacial pathology in Main military hospital during the Great Patriotic War]. AB - There was marked work of dental unit of the Main military hospital during the Great Patriotic War, including during evacuation in Gor'ky. Insertion of modern methods of immobilization of fragments of jaws and well organized succession to 1944 permitted to decrease the quantity of operations of traumatic osteomyelitis and increase the quantity of plastic operations. Owing to using of new materials (stainless steel, plastics) and to insertion of antibiotics, the period after 1946 is characterized by increased capabilities of effectuating of plastic operations and treatment of jaw breakups. PMID- 20017373 TI - [The 90th anniversary of the Center of State Sanitation-and-Epidemiological Control of Privolzhsky-Ural military region]. AB - There was exposed in brief the history of "The 1321" Center of state sanitation and-epidemiological control of Privolzhsky-Ural military region". Nowadays it's a profile institute, where high-qualified and low-grade physicians are working. The Center successfully solves the tasks of securing of sanitarium-epidemiological well-being of regional troops, realizes a grand volume of education-methodical, science-practical and info-analyzing work. PMID- 20017374 TI - [Efficiency of normobaric hypoxytherapy in the combined rehabilitation of children and adolescents with arterial hypertension]. AB - The present study included 166 children and adolescents presented with arterial hypertension. It has demonstrated high efficiency of normobaric hypoxytherapy of this condition. Its advantages over the standard electrotherapeutic sleep treatment is confirmed by the results of 24 hour arterial pressure monitoring. Criteria for the efficiency of combined rehabilitation have been developed including average day and night time arterial pressure, its maximum level, day and night hypertension time indices. PMID- 20017375 TI - [Experience with the use of ozone for the treatment of chronic viral hepatitis]. AB - Comparative data are presented illustrating efficiency of the treatment of patients with chronic viral hepatitis by standard therapeutic methods and basal therapy combined with ozone therapy. Patients treated with the use of ozone exhibited better biochemical, virological, and functional characteristics compared with their counterparts managed by standard methods. The results of the study suggest a higher efficiency of combined therapy and reduced risk of development of cholelithiasis. PMID- 20017376 TI - [Correction of syndrome-associated metabolic disturbances in patients with erosive and ulcerative lesions in the gastroduodenal system]. AB - Correction of syndrome-associated metabolic disturbances in patients with erosive and ulcerative lesions in the gastroduodenal system was achieved by inclusion of moderately mineralized drinking water (Essentuki No 4 and the like), low-sulfide mineral baths, and essential phospholipids in the system of combined sanatorium and-spa treatment. This approach allowed metabolic status of the patients to be improved and peroxide homeostasis stabilized. Moreover, it had generalized beneficial effect on the pathological process. PMID- 20017377 TI - [The use of structural resonance electromagnetotherapy and liquid synbiotics for the rehabilitative treatment following cholecystectomy]. AB - The objective of the work was to evaluate the efficiency of combined treatment using structural resonance electromagnetotherapy (SRMT) and liquid synbiotics in the early period after cholecystectomy. A total of 72 (91.7%) operated patients were found to have intestinal dysbiosis of different severity. Analysis of cardiac rhythm variability revealed relative predominance of sympathetic rhythm modulations and enhanced ergotropic component of the total spectrum power. Assessment of the quality of life showed that restricted physical activity makes the most important contribution to its impairment in the patients who underwent cholecystectomy. Protracted therapy with liquid synbiotics and concomitant structural resonance electromagnetotherapy promoted restoration and further improvement of intestinal microbiocenosis, reduction of sympathetic modulations of cardiac rhythm, and optimization of the patients' quality of life. PMID- 20017378 TI - [Comparative hemodynamic characteristics of patients with the organ of vision subjected to low-intensity laser radiation of the green and red wavelengths]. AB - This comparative study was designed to evaluate effect of low-intensity speckle laser radiation in the green and red wavelengths on intraocular hemodynamics in 202 patients (202 eyes) with retinal vascular problems and without them. Irradiation was performed using "Izumrud" and "Rubin" adapters for an AMO-ATOS unit. Peak systolic velocity and peripheral resistance index in posterior short ciliary arteries and central retinal artery were measured. It was shown for the first time that green speckle laser radiation (wavelength 535 nm) has more pronounced (compared with the red light) effect on intraocular hemodynamics; thereby, it makes a greater contribution to the improvement of the visual function (e.g. in the case of glaucoma) and to the widening of the fields of vision. PMID- 20017379 TI - [Efficiency of non-medicamentous therapy and evaluation of intersystemic relationship between functional reserves of the organism and periodontal status in patients with chronic generalized periodontitis]. AB - Mechanisms of mutual influence and aggravation of splanchnic organ and oral cavity diseases are discussed in the context of the integral anthropological approach. The systemic character of analysis of pathogenetic mechanisms of chronic generalized periodontitis dictates the necessity of an absolutely new conceptual outlook of organization and implementation of medical and preventive measures designed to activate sanogenetic responses and to enhance the regulatory potential and functional reserves of the organism in the presence of developing periodontitis. It was shown that inclusion of intermittent hypoxic training in the combined rehabilitative treatment of the patients with chronic periodontitis substantially promotes corrective influences on periodontal tissues and ensures stable clinical effect. PMID- 20017380 TI - [Pathogenetic therapy of tuberculosis of respiratory organs during sanatorium-and spa treatment]. AB - The objective of this study was to evaluate the efficiency of combined inhalation of cycloferon for 5 weeks and oral intake of cytoflavin during the period of sanatorium-and-spa treatment of the patients with pulmonary tuberculosis. It was shown that combined immunotrophic and antioxidative treatment promotes resolution of focal and infiltrative lesions in the lungs, normalization of hemograms, and minimization of the activity of cytolytic and cholestatic markers of the liver. PMID- 20017381 TI - [Approaches to optimization of the organizational and economic management of a sanatorium-and-spa facility]. AB - The main goals of reorganization of sanatorium-and-spa facilities and their activities under current economic conditions should be standardization of services provided to the patients, improvement of their quality, efficacious exploitation of the available resources, and motivation of the personnel. PMID- 20017382 TI - [Application of contrast thermopuncture for the treatment of dorsopathies]. AB - The objective of this work was to study the possibility of replacing acupuncture variants of reflexotherapy of dorsopathies by non-invasive procedures, such as thermopuncture at biologically active points. Mechanisms of action of contrast thermopuncture were elucidated. Its clinical efficiency for the rehabilitative treatment of cervical and thoracic dorsopathies was demonstrated. It is concluded that many specific features of the method account for its advantages over classical acupuncture. PMID- 20017383 TI - [Gerasimov effect of an alternating magnetic field on clinical characteristics of peripheral blood from women suffering tuboperitoneal infertility of inflammatory origin]. AB - The influence of an alternating magnetic field (AMF) on the number of erythrocytes, hemoglobin level, erythrocyte sedimentation rate, differential blood cell count, and pH values in peripheral blood was studied in 15 woman volunteers suffering tuboperitoneal infertility (TPI) of inflammatory origin. They were found to have a significantly (p < 0.05) lower number of erythrocytes, absolute number of stab neutrophils, and relative number of segmented neutrophils compared with the respective normal values. In addition, their blood pH values were decreased. The above changes are characteristic of hypoxic conditions supposed "to train" tissues for chronic inflammation; such training may underlie the mechanism of therapeutic effect of alternating magnetic field in women with tubuloperitoneal infertility. PMID- 20017384 TI - [Rehabilitation of the patients with polycystic ovary syndrome during sanatorium and-spa treatment]. AB - An original method for the combined non-medicamentous balneotherapeutic treatment of the patients with polycystic ovary syndrome and compromised reproductive function has been developed. The method designed to be applied after laparoscopic intervention for the management of infertility includes the use of radon water in combination with acupuncture. It was shown to help to restore regular menstrual cycles in half of the treated women and normalize fertility in some of them. PMID- 20017385 TI - [Application of low-power visible and near infrared radiation in clinical oncology]. AB - Although low-power visible (VIS) and near infrared (nIR) radiation emitted from lasers, photodiodes, and other sources does not cause neoplastic transformation of the tissue, these phototherapeutic techniques are looked at with a great deal of caution for fear of their stimulatory effect on tumour growth. This apprehension arises in the first place from the reports on the possibility that the proliferative activity of tumour cells may increase after their in vitro exposure to light. Much less is known that these phototherapeutic modalities have been successfully used for the prevention and management of complications developing after surgery, chemo- and radiotherapy. The objective of the present review is to summarize the results of applications of low-power visible and near infrared radiation for the treatment of patients with oncological diseases during the last 20-25 years. It should be emphasized that 2-4 year-long follow-up observations have not revealed any increase in the frequency of tumour recurrence and metastasis. PMID- 20017386 TI - [Current state-of-the-art of rehabilitative treatment of chronic obliterative arterial diseases of the lower extremities]. AB - Up-to-date approaches and methods for balneotherapy of chronic obliterative arterial diseases of the lower extremities are discussed with special reference to their confirmed clinical efficiency. The necessity of differential application of physical agents to the treatment and rehabilitation of the patients with these diseases is substantiated. PMID- 20017387 TI - [Comparison of algorithms for prediction of related proteins using the method of phylogenetic profiles]. AB - Computational interactomics deals with prediction of functionally related proteins. One approach for solving this problem using comparative genomics consists in analysis of similarities between phylogenetic profiles of proteins. In contrast to most methods, which predict only pairwise interactions between proteins, in the present work we have applied cluster analysis techniques in order to find modules of functionally related proteins. We have performed cluster analysis of phylogenetic profiles of E. coli proteins using several clustering techniques and distances between profiles. We report here, that the best correspondence in the composition of resultant clusters to known metabolic pathways is achieved using Ward's clustering together with Hamming's distance. The proposed technique of assessing predictions of the modules of functionally related proteins can be used for comparative analysis of different algorithms for computational interactomics. PMID- 20017388 TI - [Computer-assisted search and optimization of new integrase inhibitor of human immunodeficiency virus]. AB - Finding and optimization of new HIV-1 integrase inhibitors is an important task due to the drawbacks of the existing antiretroviral therapy caused by the significant adverse effects, high cost of the treatment, and emerging resistance. In this study we used computer-assisted methods for finding and optimization of new HIV-1 integrase inhibitors blocking the 3'-processing reaction. Possibility of application of the specialized version of computer program PASS for searching original inhibitors of 3'-processing reaction among the commercially available samples of chemical compounds, belonging to a new chemical class, is shown. We compared the general pharmacological profiles calculated by PASS for the sets of original integrase inhibitors discovered in the study with those for known inhibitors published in literature. It was shown that the predicted average selectivity is close for the compounds from both sets, while the predicted adverse/toxic actions are less for the original compounds. Differences of pharmacological profiles of original compounds and known from literature inhibitors of 3'-processing pointed out to the essential novelty of our substances. Significantly less deviations from the recommended values for QikProp parameters, which characterized the ADME properties, have been observed for the original compounds than for the integrase inhibitors published in literature. Thus, one may come to the conclusion that the original compounds look more prospective comparing to the earlier known inhibitors of 3'-processing. PMID- 20017389 TI - [Photodynamic therapy: search for ideal photosensitizer]. AB - Photodynamic therapy (PDT) is a minimally invasive and promising new modality to combat cancer. The method is based on selective accumulation of sensitizers within tumor cells. The high degree of selectivity offered by this modality has been applied for fluorescent diagnostic of cancer. Photosensitization of a tissue localized sensitizer generates cytotoxic reactive oxygen species as a result the selective destruction of tumor may be achieved. The PDT's major advantages compared to traditional methods of cancer treatment are better selectivity, low skin and general toxicity. This review highlights first and second generations of sensitizers, their photosensitizing abilities and drawbacks. Future developments in PDT will certainly include the discovery of new photosensitizers and a broadening of the applications of the treatment by various means. PMID- 20017390 TI - [The prospect of pluripotent stem cell-based therapy]. AB - Human embrional stem cells (hESC) are able to maintain pluripotency in culture, to proliferate indefinitely and to differentiate into any somatic cell type. Due to these unique properties, hESC may become an exceptional source of tissues for transplantation and have great potential for the therapy of incurable diseases. Here, we review new developments in the area of embrional stem cells and discuss major challenges--standartization of protocols for cell derivation and cultivation, identification of specific molecular markers, development of new aprouches for directed differentiation etc.--which remain to be settled, prior to safe and successful clinical application of stem cells. We appraise several potential approaches of hESC therapy including derivation of autologous cells via therapeutic cloning (1), generation of immune tolerance to allogenic donor cells via hematopoetic chimerism (2), and development of the banks of hESC lines (3). In addition, we discuss brifly induced pluripotent cells, which are derived via genetic modification of autologous somatic cells and are analogous to ESC. Our analysis demonstrates that uncontrollable differentiation in vivo and teratogenic potential of hESC are critical limitations of their application in clinic. Therefore, the major direction of hESC use is derivation of a specific differentiated progeny, which has lower proliferative potential and immune privilege, yet poses fewer risks. Finally, cell therapy is far more complex and resource-consuming process as compared to drug-based medicine; pluripotent stem cell biology and technology is in need of further investigation and development before these cells can be used in clinics. PMID- 20017391 TI - [Stem cells of intestine epithelium. Survival mechanisms and microbiota role]. AB - The thanatogenetic mechanisms of stem cells (SC) of rapidly renewing system of intestinal epithelium still remain unclear. On the one hand, they are definitely involved into basic mechanisms of carcinogenesis in the gastrointestinal tract, because dysregulation of programs responsible for elimination of "unwanted" mutant cells (which are normally under immune and own intrinsic control) is one of the reasons of neoplastic expansion. On the other hand, elucidation and characterization of the regulatory machinery controlling SC survival are interrelated with problems of clinical medicine, including the increase of therapeutic efficiency of treatment of inflammatory and ulcer lesions of the gut, traumatic and surgical wounds, as well as restriction of side effects in normal tissues induced by application of intensive methods chemo- and radiotherapy of cancer. The latter is especially important for treatment of blood diseases and tumors of peritoneal cavity organs mainly due to bone marrow and intestinal epithelium damage. (These tissues are the most sensitive to these treatments.) The review considers data on exogenous and genetic modifiers of SC survival, and also the basic principles of mechanisms involved into renewal and regeneration of SC and the effects of microbiota on these processes. PMID- 20017392 TI - [A new method for quantitative estimation of the number of virus particles]. AB - [In the present work virus particles of live mumps virus vaccine widely used for vaccination in Russia have been detected and visualized by atomic force microscopy. For quantitative estimation of the number of observed virus particles the special method has been proposed. The presence of protein component of the virus in vaccine was tested by ELISA and dot-blot analysis. Using quantitative real-time PCR assay the number of copies of viral RNA was estimated. The results of quantitative estimation obtained by real-time PCR corresponded with atomic force microscopy data. PMID- 20017393 TI - [The lysosomal cathepsins B, L and D in development of murine experimental leukemias]. AB - Lysosomal proteases are actively involved in pathogenesis of cancer progression. Alterations in proteases and their inhibitors interaction were suggested to be implicated in the processes of tumor invasion and metastasis. Among proteases connected with malignant growth, cysteine cathepsins B and L and aspartic cathepsin D play the main role in the tumor development. The present study was designed to investigate activity of cathepsins B, L and D activity in the development and treatment of murine experimental leukemias and to determine the correlation of these proteases with tumor malignancy and the chemotherapy effect. P-388 leukemia was characterized by a more aggressive development and unfavorable prognosis than L1210/1 leukemia. The activity of cathepsins B, L and D in tumor tissues of mice infected with P-388 leukemia, as well as in liver and spleen and the activity of cathepsins B and L in serum were lower than their activity in mice infected with L1210/1 leukemia. Changes of cathepsin activity in liver and spleen of mice with leukemias have demonstrated a level of aggressiveness of tumor development and invasion of liver and spleen by neoplastic cells. The treatment resulted in the increase of cathepsin B, L and D activities in tumor tissue, liver, spleen and cathepsin B and L activities in serum. The highest activity of proteases was revealed in the groups of mice characterized by the greatest suppression of tumor growth. These data have shown that lysosomal proteases are involved in progression of murine experimental leukemias and elimination of tumor cells in the result of treatment. Determination of the activity of cysteine and aspartic proteases can be used for evaluation of cancer diseases malignancy, their sensitivity for chemotherapy and efficiency of treatment. PMID- 20017394 TI - [Membrane and functional effects of vinpocetine and tocopherol in rats with experimental cerebral ischemia]. AB - The membrane, antioxidant and functional effects of vinpocetine and a-tocopherol have been investigated under conditions of acute experimental cerebral ischemia in rats. Vinpocetine administration decreased accumulation of lysophospholipids in brain plasma membranes. Vinpocetine also blocked accumulation of conjugated dienes (CD). alpha-Tocopherol inhibited augmentation in CD content and did not reduce the level of lysophospholipids in brain plasma membranes. Functional consequences of membrane impairments were also detected in some behavioral tests and physical capabilities. Administration of both vinpocetine and alpha tocopherol decreased manifestations of the altered parameters induced by cerebral ischemia and vinpocetine was more effective than alpha-tocopherol. PMID- 20017395 TI - [The influense of guanidine derivatives on free radical oxidation intensity and aconitase activity at development of brain ischemia-reperfusion at rats]. AB - The study of some guanidine derivatives influence on free radical oxidation intensity and aconitase activity during the development of brain ischemia reperfusion at rats has been carried out. The biochemiluminescence parameters increasing at the brain pathology changed toward normal values under the influence of N-[4-(chlorbenzoyl)benztiazol-2-yl]guanidine, N-[imino(1 piperidinyl)methyl]guanidine and N-[imino(4-morpholinyl)methyl]guanidine. The aconitase specific activity decreased in brain and blood of animals with ischemia reperfusion. Administration of guanidine derivatives during brain ischemia reperfusion development increased aconitase specific activity. These results suggest that investigated substances can play a role of neuroprotectors, preventing free radical oxidation development. PMID- 20017396 TI - [Effects of thapsigargicin on Ca2+ movements in L1210 cells permeabilized with digitonin]. AB - The effect of Thapsigargicin (TGC), a non-phorbol ester type tumor promoter, on Ca2+ movements has been investigated using L1210 mouse lymphoma cells. Ca2+ release from intact and digitonin permeabilized cells was evaluated using Fura-2 and Fura-3. TGC like Thapsigargin (TG) has the ability to discharge the intracellular Ca2+ stores and to increase intracellular free Ca2+ concentrations. TGC in a concentration dependent manner (0.16-16 nM) also inhibited cell growth and this effect was at least partially reversed by arachidonate. PMID- 20017397 TI - [The regulation of glucose-6-phosphate dehydrogenase and glycogen synthase activities by insulin superfamily peptides in myometrium of pregnant women and its impairments under different types of diabetes mellitus]. AB - The regulatory effects of insulin, insulin-like growth factor 1 (IGF-1), and relaxin on glucose-6-phosphate dehydrogenase (G6PDH) and glycogen synthase (GS) activities have been studied in myometrium of pregnant women of control group and with diabetes mellitus of different etiology. In patients with type 1 diabetes G6PDH activity did not differ from the control group, but the enzyme activity was sharply decreased in pregnant women with type 2 diabetes and gestational diabetes. In the control group maximal stimulation of G6PDH activity was observed at 10(-9) M of peptides and their stimulating effect decreased in the following order: insulin > relaxin > IGF-1. In pregnant women with types 1 diabetes insulin effect on the enzyme activity was lower than in the control, and the effects of IGF-1 and relaxin were absent. In the group of pregnant women with type 2 diabetes and gestational diabetes the effects of insulin and IGF-1 were decreased, but the effect of relaxin was somewhat higher thus giving the following order in their efficiency relaxin > IGF-1 = insulin. At 10(-9) M peptides exhibited similar stimulating effects on the active form of GS-I, but had no influence on the total enzyme activity in the control group of pregnant women. In patients with type 1 diabetes GS activity remained unchanged (versus control), and peptides did not stimulate the enzyme activity. In patients with type 2 diabetes a significant decrease in GS activity was accompanied by the decrease in the effect of peptides, giving the following order of their efficiency: insulin = IGF-1 > relaxin. In myometrium of pregnant women with gestational (treated and untreated) diabetes GS activity decreased, the effect of insulin was weaker, whereas the effects of relaxin and IGF-1 increased thus giving the following order of their efficiency: relaxin > IGF-1 > insulin. Insulin therapy of type 1 diabetes incompletely restored sensitivity of the enzymes to the peptide actions. At the same time, in women with gestational diabetes and subjected to insulin therapy the stimulating effect of relaxin on the enzyme activities increased. This fact suggests that relaxin exhibits replacement functions under conditions of attenuated insulin action. PMID- 20017398 TI - [Proteolytic activity of placenta with EPH-gestosis]. AB - Essential edema-proteinuria-hypertension (EPH) gestosis still represents an important obstetrical problem. We have investigated the activity of carboxypeptidase H (CPH), phenylmethylsulfonyl fluoride inhibited carboxypeptidase (PMSF-CP), carboxypeptidase M (CPM) and angiotensin-converting enzyme (ACE), the main carboxypeptidases in human placenta under normal conditions and mild EPH-gestosis. Gestosis was accompanied by the decrease in activity of the enzymes involved into metabolism of regulatory peptides (ACE, CPH, PMSF-CP, CPM) compared with their activity in placenta under physiological pregnancy. Correlation analysis revealed positive correlation between placental CPH and CPM (r = 0.2735*) in EPH-gestosis. These findings suggest involvement of placental proteases into formation of compensatory-adaptive reactions in the fetoplacental complex at EPH-gestosis; the data obtained may be also employed for the development of methods of prophylaxis and corrections of metabolic impairments in pathology of pregnancy. PMID- 20017399 TI - [Regulation of progenitor cell function by hyaluronidase]. AB - This study demonstrated the possibility of in vivo activation of progenitor cells by hyaluronidase. Specifically, treatment with hyaluronidase increased the number of mesenchymal and bone marrow precursor cells, their proliferative activity, and differentiation. Also, it promoted stem cell mobilization into blood under effect of granulocyte colony-stimulating factor (G-CSF) and enhanced progenitor cell adhesive properties. Therapeutic efficiency of transplantation of mononuclear cells isolated from peripheral blood after administration of G-CSF improved under effect of hyaluronidase. Hyaluronidase immobilized by electron beam synthesis nanotechnology exhibited high specific activity with respect to stem cells via both enteral and parenteral routes. PMID- 20017400 TI - [New promising natural immunocorrective agents]. AB - The influence of Sorbus sibirica, Calendula officinalis and Althaea officinalis extracts on the humoral immune response and nonspecific resistance of mice to immunosuppression by cyclophosphan was studied. It was shown that these extracts are not inferior to Echinacea purpurea tincture in terms of stimulation of humoral immune response, phagocytic and bactericidal activity of peritoneal macrophages but exceed effect of E. purpurea on phagocytic activity of peripheral blood neutrophils. PMID- 20017401 TI - [Neuroprotective effects of dibornol and mechanism of its action in rats with cerebral ischemia]. AB - Neuroprotective activity of the new sterically hindered phenolic antioxidant 4 methyl-2,6-diisobornylphenol (dibornol) in rats with total transient cerebral ischemia was investigated. Dibornol decreased mortality of rats and the number of animals with severe neurological deficit; moreover, it accelerated restoration of neurological status in the survived rats. Neuroprotective activity of dibornol is based on its ability to diminish lipid peroxidation in ischemic brain, suppress cerebral tissue hypoxia and protect functional activity of endothelium. Improved oxygen delivery was a consequence of reduced hyperviscosity syndrome (enhanced deformability of erythrocytes and their decreased aggregation). PMID- 20017402 TI - [Hepatotoxicity of antineoplastic agents]. AB - The influence of various antineoplastic preparations (farmorubicin, paclitaxel, etoposide, platidiam) on morphological and functional status of the liver in experimental animals was studied. It is shown, that all these antineoplastic drugs possess strong hepatotoxic activity and thereby cause toxic hepatitis and increase activity of hepatic enzymes. Biochemical parameters were normalized within 30 days after treatment whereas morphological changes persisted somewhat longer. PMID- 20017403 TI - [Prospects for pharmacological regulation of macrophage activity by modulation of intracellular signal cascade]. AB - This article is devoted to classical and alternative macrophage activation, and intracellular transmission of external signals onto genes. It presents data on postreceptor events resulting in the formation of classical or alternative properties of macrophages. The role of some molecules of intracellular signaling pathways (STAT1, NF-kappaB, IRAK, TRAF6, Jak1, Tyk2, STAT3, c-Maf, Sp1, C/EBP, CREB, STAT6, MAP-kinase, PI3-kinase, c P) in the development of proinflammatory and anti-inflammatory activities of macrophages is discussed. PMID- 20017404 TI - [Influence of taxan-containing cytostatic preparations on the progeny]. AB - In experiments designed to study effect of taxan-containing agents on the progeny of Wistar rats, one of the parents was given a single intravenous injection of the antineoplastic drug paclitaxel 1, 3 and 6 months before crossing with the untreated partner. The number of systemic pathological changes in the off-springs of paclitaxel-treated females was significantly greater and their spectrum wider than in those of the treated males. Severity of toxic effects depended on the time of crossing after drug administration. The optimal time for obtaining less affected off-springs of treated females and males was 3 and 6 months after administration of paclitaxel. PMID- 20017405 TI - [Evaluation of anti-inflammatory activity of extracts from Siberian plants]. AB - Experimental investigations have shown that water-alcohol extracts from plants containing alkaloids (Aconitum baikalense, Aconitum septentrionale, Delphinium elatum L., Conium maculatum) and salicylic acid (Filipendula ulmaria, Salix viminalis, Fragaria vesca, Rubus idaeus) inhibited the development of main symptoms of inflammation, viz. exudation, pain, fever, to the same extent as non steroidal anti-inflammatory agents. The substances studied in this work may be used to develop new efficient pharmacological preparations for the treatment of different inflammatory conditions associated with severe pain syndrome. PMID- 20017406 TI - [Anti-ulcerative effect of non-starch polysaccharides]. AB - Standard models of experimental ulceration (of neurogenic origin, H. Shay ulcer, indomethacine-, ethanol-, prednisolone-, histamine- and acetate-induced ulcers) were used to demonstrate protective effect of non-starch polysaccharides (potassium alginate, potassium pectate, low-esterified pectin). Potassium pectate proved to be the most efficacious protector. Mechanism of its anti-ulcerative action is attributable to antacidic, cytoprotective, and reparative activity. It appeals to optimally stimulate the motor-evacuation function and, besides, exhibits marked anti-inflammatory activity. PMID- 20017407 TI - [Chitotriosidase of human macrophages and mammalian chitinases: biological functions and abnormalities in pathology]. AB - Recently discovered human chitotriosidase is a member of chitinase family capable to hydrolyze chitin, a polymer of N-acetylglucosamine. Serum and plasma chitotriosidase activity is usually measured as the first step in diagnosis of Gaucher disease. Monitoring chitotriosidase activity is widely used during treatment of this pathology by enzyme replacement therapy. Its elevated plasma level reflects gradual intralysosomal accumulation in Gaucher cells (lipid-loaded macrophages). Macrophages overloaded by the enzyme accumulated in lysosomal material (lipids) were shown to secrete chitotriosidase; its increased expression was noted in several lysosomal storage diseases and atherosclerosis. We observed enhanced chitotriosidase activity in healthy young and elderly persons probably associated with the development of atherosclerosis and plaque formation. Chitotriosidase is believed to play a role in mechanisms of immunity and protection against chitin-containing pathogens. PMID- 20017408 TI - [Quantitative assessment of in vitro neutralizing activity of secretory factors from rat lungs against influenza virus]. AB - Secretory factors were isolated by lung wash followed by centrifugation to remove cells, dialysis of supernatant to remove NaCl salt, lyophilization of the lavage fluid and resuspention of the lyophilization product in an isotonic NaCl solution. It was shown that biological activity of influenza virus /Aichi/2/68 (3N2) significantly decreased (p = 0,01) from 8,17 +/- 0,10 to 7,14 +/- 0,20 IgEID50/ml during its incubation with secretory factors at 37 degrees C for 1 hr and to 7,92 +/- 0,17 IgEID50/ml in isotonic NaCl solution in the absence of these factors. Their concentration in the incubation medium was estimated to be 9.1 +/- 0.7% of their level in the lungs. PMID- 20017409 TI - Shelly Spiro: a pharmacy technology guru ASCP president 2009-2010. AB - Rachelle "Shelly" Spiro, RPh, FASCP, incoming president of the American Society of Consultant Pharmacists (ASCP), believes sharing information electronically will greatly benefit pharmacy practice. In recent years she has been an expert in health care technology, helping establish standards for reporting health care data on outcomes, reimbursement, and electronic medical records--all areas that affect consultant pharmacists. One of her goals as president is to help mentor potential leaders by having a grassroots campaign to reach out to students in schools of pharmacy. In addition, she wants ASCP's leadership to move to a more regional structure and get members more involved locally at state levels. PMID- 20017410 TI - Assessment of the use of xerogenic medications for chronic medical and dental conditions among adult day health participants. AB - OBJECTIVES: To describe the health conditions, dental problems, and use of xerogenic medications among dental patients in adult day health (ADH) centers. DESIGN: Cross-sectional descriptive study. SETTING: ADH centers in King County, Washington. POPULATION: ADH clients who were patients of a mobile dental service. INTERVENTIONS: Pharmacist-conducted chart reviews and in-person medication reviews with patients. MAIN OUTCOME MEASURES: Demographic description, mean numbers of medical and dental problems, medications, xerogenic medications used per subject, and identification of xerogenic medications by therapeutic class. RESULTS: At five sites, 97 patients were interviewed (average age 73.8 +/- 11.8 years, 61% female); ethnicities included: Asian-American (37.1%), Caucasian (30.9%), Russian (29%), and African-American (3%). Mean numbers of chronic health problems, medications, and xerogenic medications per patient were 5.2 +/- 2.7, 10.9 +/- 4.4, and 3.3 +/- 1.8, respectively. Antidepressants were the most commonly used xerogenic medication, followed by antipsychotics, antiemetics, analgesics, and antihistamines. Among 74 patients who received dental treatment, 33 (44.6%) wore dentures. Among 58 patients with teeth, a mean number of 2.8 dental problems per patient was identified. Dental caries (51.7%) was the most prevalent problem, followed by periodontitis (29.3%), soft tissue lesions (10.3%), gingivitis (5.2%), and candidiasis (3.4%). CONCLUSION: Multiple systemic diseases, use of multiple xerogenic medications, and poor oral health were prevalent among the ADH clients in this study. However, self-reports of dry mouth were unrelated to number of xerogenic medications or oral conditions. Further research is needed to determine the association between self-reported dry mouth, chronic health conditions, use of xerogenic medications, tooth loss, and/or denture use. PMID- 20017411 TI - Recurrent episodes of serotonin-reuptake inhibitor-mediated hyponatremia in an elderly patient. AB - OBJECTIVE: To report a case of recurrent episodes of serotonin-reuptake inhibitor mediated hyponatremia in an elderly patient and compare it with other reports of similar occurrences. CASE SUMMARY: A 66-year-old white woman was diagnosed with drug-induced syndrome of inappropriate antidiuretic hormone (SIADH) attributed to selective serotonin-norepinephrine-reuptake inhibitor (SNRI) therapy. Duloxetine was initiated following failure of several other antidepressants. The patient was admitted with sudden onset altered mental status, memory loss, personality changes, and a serum-sodium level of 128 mEq/L (range 135-145 mEq/L), despite receiving sodium supplementation. The duloxetine dose was 60 mg daily. Three months later she presented to the emergency department with complaints of lethargy, muscle weakness, nausea, altered mental status, and a serum sodium level of 129 mEq/L. SIADH was diagnosed and attributed to duloxetine therapy. Duloxetine was titrated to 30 mg every other evening. She remained stable on the lower dose, fluid restriction, and sodium supplementation. Diuretic reinitiation and sodium supplementation discontinuation resulted in serum sodium of 123 mEq/L. This increased to low/normal (136 mEq/L) with duloxetine discontinuation. A rechallenge with escitalopram resulted in low serum-sodium levels. DISCUSSION: A PubMed search was done utilizing the terms duloxetine, elderly, hyponatremia, selective serotonin-reuptake inhibitor, SSRI, SNRI, syndrome of inappropriate antidiuretic hormone, SIADH, and selective serotonin-norepinephrine reuptake inhibitor to find similar reports. CONCLUSION: Clinicians evaluating older patients taking serotonin-reuptake inhibitors are encouraged to monitor serum sodium if the patient presents with vague, nonspecific symptoms commonly associated with older age or depression to rule-out SIADH. PMID- 20017412 TI - Pharmacological management of benign prostatic hypertrophy with phosphodiesterase 5 inhibitors. AB - Benign prostatic hypertrophy (BPH) and its associated lower urinary-tract symptoms (LUTS) commonly result in a significant decrease in the quality of life (QOL) of aging men. Although several safe and effective medications are currently available for the management of this disease, newly emerging research has focused on the exploration of unconventional pharmacological therapy. In particular, multiple clinical trials published within the past two years have explored the use of phosphodiesterase-5 (PDE-5) inhibitors for the alleviation of LUTS secondary to BPH. Although this class of medication has traditionally been used in men with erectile dysfunction (ED), the preliminary results from these studies have offered promising evidence for use of PDE-5 inhibitors in BPH. PMID- 20017413 TI - Genotoxicity of 1,3-butadiene and its epoxy intermediates. AB - Current risk assessments of 1,3-butadiene (BD*) are complicated by limited evidence of its carcinogenicity in humans. Hence, there is a critical need to identify early events and factors that account for the heightened sensitivity of mice to BD-induced carcinogenesis and to deter-mine which animal model, mouse or rat, is the more useful surrogate of potency for predicting health effects in BD exposed humans. HEI sponsored an earlier investigation of mutagenic responses in mice and rats exposed to BD, or to the racemic mixture of 1,2-epoxy-3-butene (BDO) or of 1,2,3,4-diepoxybutane (BDO2; Walker and Meng 2000). In that study, our research team demonstrated (1) that the frequency of mutations in the hypoxanthine-guanine phosphoribosyl transferase (Hprt) gene of splenic T cells from BD-exposed mice and rats could be correlated with the species-related differences in cancer susceptibility; (2) that mutagenic-potency and mutagenic specificity data from mice and rats exposed to BD or its individual epoxy intermediates could provide useful information about the BD metabolites responsible for mutations in each species; and (3) that our novel approach to measuring the mutagenic potency of a given chemical exposure as the change in Hprt mutant frequencies (Mfs) over time was valuable for estimating species specific differences in mutagenic responses to BD exposure and for predicting the effect of BD metabolites in each species. To gain additional mode-of-action information that can be used to inform studies of human responses to BD exposure, experiments in the current investigation tested a new set of five hypotheses about species-specific patterns in the mutagenic effects in rodents of exposure to BD and BD metabolites: 1. Repeated BD exposures at low levels that approach the occupational exposure limit for BD workers (set by the U.S. Occupational Safety and Health Administration) are mutagenic in female mice. 2. The differences in mutagenic responses of the Hprt gene to BD in similarly exposed rodents of a given species (reported in various earlier studies) are primarily associated with age-related thymus activity and trafficking of T cells and with sex-related differences in BD metabolism. 3. The mutagenic potency of the stereochemical forms of BD's epoxy intermediates plays a significant role in the species-related mutagenicity of BD. 4. The hydrolysis-detoxification pathway of BD through 1,2-dihydroxy-3-butene (BD-diol) is a major contributor to mutagenicity at high-level BD exposures in mice and rats. 5. Significant and informative species-specific differences in mutation spectra can be identified by examining both large- and small-scale genetic alterations in the Hprt gene of BD exposed mice and rats. The first four hypotheses were tested by exposing mice and rats to BD, meso-BDO2, or BD-diol and measuring Hprt Mfs as the primary biomarker. For this, we used the T-cell-cloning assay of lymphocytes isolated from the spleens of exposed and control (sham-exposed) mice and rats. The first hypothesis was tested by exposing female B6C3F1 mice (4 to 5 weeks of age) by inhalation for 2 weeks (6 hours/day, 5 days/week) to 0 or 3 ppm BD. Hprt Mfs were measured at the time of peak mutagenic response after exposure for this age of mice. We then compared the resulting data to those from mutagenicity studies with mice of the same age that had been exposed in a similar protocol to higher levels of BD (Walker and Meng 2000). In mice exposed to 3 ppm BD (n = 27), there was a significant 1.6-fold increase over the mean background Hprt Mf in control animals (n = 24, P = 0.004). Calculating the efficiency of Hprt mutant induction, by dividing induced Hprt Mfs by the respective BD exposure levels, demonstrated that the mutagenic potency of 3 ppm BD was twice that of 20 ppm BD and almost 20 times that of 625 or 1250 ppm BD in exposed female mice. Sample-size calculations based on the Hprt Mf data from this experiment demonstrated the feasibility of conducting a future experiment to find out whether induced Mfs at even lower exposure levels (between 0.1 and 1.0 ppm BD) fit the supralinear exposure response curve found with exposures between 3.0 and 62.5 ppm BD, or whether they deviate from the curve as Mf values approach the background levels found in control animals. The second hypothesis was tested by estimating mutagenic potency for female mice exposed by inhalation for 2 weeks to 0 or 1250 ppm BD at 8 weeks of age and comparing this estimate to that reported for female mice exposed to BD in a similar protocol at 4 to 5 weeks of age (Walker and Meng 2000). For these two age groups, the shapes of the mutant splenic T-cell manifestation curves were different, but the mutagenic burden was statistically the same. These results support our contention that the disparity in responses reported in earlier Hprt mutation studies of BD-exposed rodents is related more to age-related T-cell kinetics than to age-specific differences in the metabolism of BD. The third hypothesis was tested by estimating mutagenic potency for female mice and rats (4 to 5 weeks of age) exposed by inhalation to 2 or 4 ppm meso-BDO2 and comparing these estimates to those previously obtained for female mice and rats of the same age and exposed in a similar protocol to (+/-)-BDO2 (Meng et al. 1999b; Walker and Meng 2000). These exposures to stereospecific forms of BDO2 caused equivalent mutagenic effects in each species. This suggests that the small differences in the mutagenic potency of the individual stereoisomers of BDO2 appear to be of less consequence in characterizing the sources of BD-induced mutagenicity than the much larger differences between the mutagenic potencies of BDO2 and the other two BD epoxides (BDO and 1,2-dihydroxy-3,4-epoxybutane [BDO-diol]). The fourth hypothesis was tested in several experiments. First, female and male mice and rats (4 to 5 weeks of age) were exposed by nose only for 6 hours to 0, 62.5, 200, 625, or 1250 ppm BD or to 0, 6, 18, 24, or 36 ppm BD-diol primarily to establish BD and BD-diol exposure levels that would yield similar plasma concentrations of BD-diol. Second, animals were exposed in inhalation chambers for 4 weeks to 0, 6, 18, or 36 ppm BD-diol to determine the mutagenic potency estimates for these exposure levels and to compare these estimates with those reported for BD-exposed female mice and rats (Walker and Meng 2000) in which similar blood levels of BD diol had been achieved. Measurements of plasma concentrations of BD-diol (via a gas chromatography and mass spectrometry [GC/MS] method developed for this purpose) showed these results: First, BD-diol accumulated in a sublinear manner during a single 6-hour exposure to more than 200 ppm BD. Second, BD-diol accumulated in a linear manner during single (6-hour) or repeated (4-week) exposure to 6 or 18 ppm BD and in a sublinear manner with increasing levels of BD diol exposure. Third, exposure of female mice and rats to 18 ppm BD-diol produced plasma concentrations equivalent to those produced by exposure to 200 ppm BD (exposure to 36 ppm BD-diol produced plasma concentrations of about 25% of those produced by exposure to 625 ppm BD). In general, 4-week exposure to 18 or 36 ppm BD-diol was significantly mutagenic in female and male mice and rats. The differences in mutagenic responses between the species and sexes were not remarkable, except that the mutagenic effects were greatest in female mice. The substantial differences in the exposure-related accumulation of BD-diol in plasma after rodents were exposed to more than 200 ppm BD compared with the relatively small differences in the mutagenic responses to direct exposures to 6, 18, or 36 ppm BD-diol in female mice provided evidence that the contribution of BD-diol derived metabolites to the overall mutagenicity of BD has a narrow range of effect that is confined to relatively high-level BD exposures in mice and rats. This conclusion was supported by the results of parallel analyses of adducts in mice and rats concurrently exposed to BD-diol (Powley et al. 2005b), which showed that the exposure-response curves for the formation of N-(2,3,4 trihydroxybutyl)valine (THB-Val) in hemoglobin, formation of N7-(2,3,4 trihydroxybutyl)guanine (THB-Gua) in DNA, and induction of Hprt mutations in exposed rodents were remarkably similar in shape (i.e., supralinear). Combined, these data suggest that trihydroxybutyl (THB) adducts are good quantitative indicators of BD-induced mutagenicity and that BD-diol-derived BDO-diol (the major source of the adducts) might be largely responsible for mutagenicity in rodents exposed to BD-diol or to hight levels of BD. The mutagenic-potency studies of meso-BDO2 and BD-diol reported here, combined with our earlier studies of BD, (+/-) BDO, and(+/-)-BDO2 (Walker and Meng 2000), revealed important trends in species-specific mutagenic responses that distinguish the relative degree to which the epoxy intermediates contribute to mutation induction in rodents at selected levels of BD exposures. These data as a whole suggest that , in mice, BDO2 largely causes mutations at exposures less than 62.5 ppm BD and that BD-diol derived metabolites add to these mutagenic effects at higher BD exposures. In rats, it appears that the BD-diol pathway might account for nearly all the mutagenicity at the hight-level BD exposures where significant increases in Hprt Mfs are found and cancers are induced. Additional exposure-response studies of hemoglobin and DNA adducts specifics to BDO2, BDO-diol, and other reactive intermediates are needed to determine more definitively the relative contribution of each metabolite to the DNA alkylation and mutation patterns induced by BD exposure in mice and rats. For the fifth hypothesis, a multiplex polymerase chain reaction (PCR) procedure for the analysis of genomic DNA mutations in the Hprt gene of mice was developed. (ABSTRACT TRUNCATED) PMID- 20017414 TI - The Nobel Prize in Physiology or Medicine 2009. PMID- 20017415 TI - [Acute renal failure in premature neonates]. AB - BACKGROUND/AIM: Hemodynamic stress is the leading cause of acute renal failure (ARF) in premature neonates. Incidence of ARF in this population is between 8 and 24%. The aim of this study was to determine the frequence of presence of ARF in premature neonates, as well as its impact on their survival. METHODS: A retrospective study of 114 premature neonates [(gestational age, GA less than 37 gestation weeks (gw)] admitted to the Intensive Care Unit (ICU) at the Pediatric Clinic, Institute of Child and Youth Healthcare of Vojvodina in 2007 was conducted. Serum creatinine, urea and bilirubine were determined on the 3rd day of life in 65 newborns who met inclusion criteria. ARF was diagnosed in 16 newborns (n = 16/65; 25%). RESULTS: The premature neonates with ARF had significantly lower GA [<28 gw - 8/16 (50%) vs. 5/49 (10%); p < 0.05], birth weight (BW) (1265 g vs. 1615 g; p < 0.05) and systolic blood pressure (43.37 mm Hg vs. 52.7 mmHg; p < 0.05) than ones without ARF. Non-olyguric ARF was diagnosed in 62% of newborns with ARF (n = 10/16), while the rest had the olyguric type (n = 6/16; 38%). Twenty-five percent of premature neonates with ARF (n = 4/16) died in contrast to 10% of premature neonates without ARF (n = 5/49). ARF was treated conservatively in all but 3 cases when peritoneal dialysis was performed. Renal function has recovered completely in all of the survivors. In order to determine their predictivity in relation to ARF, following parameters were analyzed: GA, BW < 1500 g, presence of concomitant sepsis and intracranial hemorrhage grade III/IV. BW < 1500 g demonstrated the highest sensitivity (se 0.75), while GA < 28 gw, sepsis and intracranial hemorrhage grade III/IV showed high specificity (sp = 0.90, 0.89 0.88, respectively). CONCLUSION: Acute renal failure frequently occurs in population of premature neonates and requires meticulous fluid and electrolyte balance, especially in the case of low birth weight and extreme immaturity. PMID- 20017416 TI - Nosocomial infections prevalence study in a Serbian university hospital. AB - BACKGROUND/AIM: Nosocomial infections (NI) are a serious health problem resulting in an enromous burden of excess morbidity and mortaliti rates, and health care costs. The aim of this study was to assess the prevalence of NI and to identify groups of patients at special risk for NI in the University Clinical Center, Kragujevac, Serbia. METHODS: A period prevalence study design was used in this study. A survey of NI included all patients hospitalized in all departments in the University Clinical Center, Kragujevac. RESULTS: Among 764 patients surveyed, the global prevalence rate of patients with at least one NI was 6.2% (95% CI = 5.6-6.8), while the prevalence of NI was 7.1%. The most frequent infections were surgical site infections (14.1%; 95% CI = 12.9-15.3), followed by pneumonia (2.3%; 95% CI = 2.1-2.5) in surgical patients. In medical wards, the most common NI were skin and subcutaneous tissue infections (1.6%; 95% CI = 1.4-1.8), and urinary infections (1.4%; 95% CI = 1.3-1.5). Overall, 85.1% NI were culture proven; the leading pathogens were Pseudomonas species (40.0%), followed by Staphylococcus species (25.0%), Escherichia coli (22.5%), Proteus mirabilis (17.5%) and Klebsiella-Enterobacter (12.5%). Multivariate logistic regression analysis identified 3 risk factors independently associated with NI appearance: hospital stay > or =8 days (p = 0.0015), urinary catheter (p = 0.0022) and antibiotic use (p < 0.001). CONCLUSION: This study showed that NI are a serious health problem in our hospital. The most common infections were surgical site infections, followed by skin and subcutaneous tissue infection and urinary tract infections. Nosocomial infections were most common in patients in urological and orthopedic departments, and then in intensive care units. Prolonged hospital stay, urinary catheter and antibiotic exposure were risk factors independently associated with NI appearance. PMID- 20017417 TI - [Risk factors for brain metastases after definitive chemoradiation for locally advanced non-small cell lung cancer]. AB - BACKGROUND/AIM: As therapy for locally advanced nonsmall cell lung carcinoma (NSCLC) improves, brain metastases (BM) still remain a great problem. The aim of the study was to analyze risk factors for BM in patients with locally advanced NSCLC after chemoradiation therapy. METHODS: Records for 150 patients with non resectable stage IIIA/IIIB NSCLC treated with combined chemoradiation therapy were analyzed. All of them had negative brain metastases imaging result before the treatment. Incidence of BM was examined in relation to age, sex, histological type, stage, performance status scale of wellbeing of cancer patients, weight loss, chemotherapy regimen and chemotherapy timing. RESULTS: One- and 2-year incidence rates of BM were 19 and 31%, respectively. Among pretreatment parameters, stage IIIB was associated with a higher risk of BM (p < 0.004) vs. stage IIIA. Histologically, the patients with nonsquamous tumors had an exceptionally high 2-year BM risk rate of 32% (p < 0.02). Examining treatment related parameters, 1-year and 2-year actuarial risk of BM were 27 and 39%, respectively, in the patients receiving chemotherapy before radiotherapy and 15 and 20%, respectively, when radiotherapy was not delayed (p < 0.03). On multivariate analysis, timing of chemotherapy (p < 0.05) and stage IIIA vs. IIIB (p < 0.01) remained statistically significant. CONCLUSION: Patients with IIIB stage, nonsquamous NSCLC, particularly those receiving sequential chemotherapy, had significantly high BM rates. PMID- 20017418 TI - [Cardiac troponin as biochemical marker of perinatal asphyxia and hypoxic myocardial injury]. AB - BACKGROUND/AIM: Myocardial cell lesion in newborns may be clinically occult. In recent years there has been shown growing interest in the use of cardiac troponin I (cTnI) in relation to perinatal asphyxia and hypoxic myocardial lesion. The aim of this study was to determine a relationship between high cTnI levels and outcome in critically ill newborns with perinatal asphyxia. METHODS: In this study 78 patients were divided into three groups. The group I included 39 newborns (15 term and 24 preterm) with perinatal asphyxia, with no deaths, only full or partial (with some neurological sequels) recovery. The group II included 10 newborns (6 preterm and 4 term), with perinatal asphyxia who died, with critical cardio-respiratory problems and multiorgan dysfunction. The group III included 29 healthy term newborns. A level of cTnI in all three groups was measured within 24-48 hours after delivery. RESULTS: A statistically significant higher value of cTnI (0.082 microg/l +/- 0.166) was found in group I than in the group III (healthy newborns). In the group I, 21/39 newborns required respiratory and 16/39 required pressure support. In the group II, the largest average value of cTnI of 0.425 +/- 0.307 was found. All of the newborns in the group II required respiratory and pressure support. In the group III the lowest average value of cTnI (0.0186 microg/L +/- 0.0286) was found. CONCLUSIONS: High cTnI levels could be used as markers of perinatal asphyxia and even as predictors of future outcomes and/or mortality. PMID- 20017419 TI - [Relationship between the type of atrial fibrillation and thromboembolic events]. AB - BACKGROUND/AIM: Atrial fibrillation (AF) increases the risk for ischemic stroke and other thromboembolic (TE) events. Aim of the study was to examine the relationship between clinical types of atrial fibrillation (AF) and (TE) events. METHODS: This longitudinal, observational study included patients with nonvalvular AF as main indication for in-hospital and/or outpatient treatment in the Cardiology Clinic, Clinical Center of Serbia during a period 1992-2007. The treatment of AF was based on the International Guidelines for diagnosis and treatment of AF, correspondent to given study period. Clinical types of AF were defined according to the latest ACC/AHA/ESC Guidelines for AF, from 2006. Diagnosis of central and systemic TE events during a follow-up was made exclusively by the neurologist and vascular surgeon. RESULTS: During a follow-up of 9.9 +/- 6 years, TE events were documented in 88/1 100 patients (8%). In the time of TE event 46/88 patients (52.3%) had permanent AF. The patients with permanent AF were at baseline significantly older and more frequently had underlying heart disease and diabetes mellitus. Cumulative TE risk during follow up was similar for patients with paroxysmal and permanent AF, and significantly higher as compared to TE risk in patients with persistent AF. However, multivariate Cox proportional hazard regression analysis with independent variables clinical types of AF at baseline and in the time of TE event, clinical and echocardiographic characteristics and therapy for prevention of TE complications at baseline and at the time of TE event, did not reveal independent predictive value of clinical type of AF for the occurrence of TE events during a follow-up. CONCLUSION: TE risk in patients with AF does not depend on clinical type of AF. Treatment for prevention of TE events should be based on the presence of well recognized risk factors, and not on the clinical type of AF. PMID- 20017420 TI - [Analysis of enzyme activity and the level of malondialdehyde in the saliva of children with gingivitis]. AB - INTRODUCTION/AIM: By analysing activity of some of the enzymes normally present in the saliva and the level of malondialdehyde in gingivitis, it is possible to estimate the functional condition of parodontium, and the examined parametres can be considered as biochemical markers of its functional condition. The aim of this paper was to examine activity of alanine aminotransferase, aspartate aminotransferase, gamma glutamyl transferase, lactate dehydrogenase and the level of malondialdehyde in the saliva of children affected with gingivitis, as well as the values of the mentioned parametres in relation to the level of the inflammation of gingiva. METHODS: The research included 120 children at the age of 12.2 with permanent dentition. Loe and Silness gingival index was used to estimate the condition of gingiva, based on which the childen were classified into four groups: the children with healthy gingiva (the control groups), the children with mild, moderate and severe inflammation of gingiva (the study group). Enzymes of the saliva were determined by the use of original tests and measured by the autoanalyser (Bio Systems A25, Spain). A modified method with tiobarbituric acid was used to determine malondialdehyde in nonstimulated mixed saliva. RESULTS: The results of the examined enzyme activity and the level of malondialdehyde in the saliva of the study groups showed statistically considerably higher values for the level of malondialdehyde (p < 0.001), for the activity of aspartate aminotransferase and gamma glutamyl transferase (p < 0.01), as well as for alanine aminotransferase (p < 0.05) in comparison with the control group, whereas the activity of lactate dehydrogenase did not show a statistically significant increase. In relation to the level of the inflammation of gingiva, the results of the examination of the enzyme activity in the study groups showed statistically significantly higher values in the group with severe inflammation in comparison with those with mild, as well as the moderate inflamatiory, except for the gamma glutamyl transferase, and in the group with moderate inflammation compared to that with the mild one, except for alanine aminotransferase. The results of the examination of the level of malondialdehyde in the saliva of the study groups did not show a statistically significantly increase in relation to the level of the inflammation of gingiva. CONCLUSION: There is a higher level of alanine aminotransferase, aspartate aminotransferase, gamma glutamyl transferase and lactate dehydrogenase enzyme activity together with the higher level of malondialdehyde in the saliva of children with gingivitis in comparison with the activity of the same enzymes and the level of malondialdehyde in the saliva of children without gingivitis. The activity of the examined enzymes in the saliva of children with gingivitis increases in relation to the intensity of the pathological process, whereas the level of malondialdehyde shows no significant difference in relation to the level of the inflammation of gingiva. PMID- 20017421 TI - [Diagnostics and treatment of ischemic heart disease in hemodialysis patients]. PMID- 20017422 TI - [Successful treatment of polymedicamentous poisoning with metoprolol, diltiazem and cilazapril]. AB - INTRODUCTION: Poisoning caused by drugs with cardiodepressive effects is an urgent condition in medicine which is associated with high mortality rate regardless of modern therapeutic methods. Accidental or intentional poisoning whit these drugs produces heart activity depression and cardiovascular collapse as consequences. Current therapy for severe poisoning caused by beta-blockers and calcium channel blockers includes both unspecific and specific antidote therapy whit glucagon, as well as application of adrenergic drugs, calcium, phosphodiesterase inhibitors and hyperinsulinemia/euglycemia therapy. However, even whit the application of these drugs, prompt measures of unspecific detoxication therapy and cardiopulmonary reanimation are crucial for survival of patients with severe poisoning. CASE REPORT: A 28-year-old female patient was hospitalized for cardiogenic shock and altered state of conscioussnes (Glasgow coma score = 4), caused by acute poisoning with 2 g of metoprolol (Presolol), 1.8 g of diltiazem (Cortiazem) and 50 mg of cilazapril (Zobox). Prolonged cardiopulmonary resuscitation was applied during the first 16 hours of hospitalization, including administration of crystaline solutions (8 L), 17 mg of adrenaline, 4 mg of atropine, 4 mg of glucagone and 1.6 g of dopamine, with electro-stimulation by temporary pacemaker and mechanical ventilation. In a defined time period, normalized state of consciousness was registered, mechanical ventilation was stopped and normal heart activity and hemodynamic stability were accomplished. During hospitalization the patient was treated for mild pneumonia and after ten days, completely recovered, was released and sent to home treatment. CONCLUSION: Prompt measures of cardiopulmonary resuscitation and multidisciplinary treatment in intensive care units significantly increase the chances of complete recovery of a patient with severe poisoning caused by drugs with cardiodepressive efects. PMID- 20017423 TI - Giant dumbbell tumor of the posterior mediastinum. AB - BACKGROUND: Giant dumbbell-shaped tumors are very rare and characterized by intra and extraspinal propagation of different dimensions. In thoracal localization, invasive growth can lead up to rib and vertebra erosion. CASE REPORT: We presented a 54-year-old woman with a giant dumbbell schwannoma in the posterior mediastinum. The tumor was removed by the posterior approach with hemilaminectomy and costotransversectomy. By microscopic examination the diagnosis of benign schwannoma was made. A year after the surgery, the patient was without neurological deficiency and without radiological signs of illness relapse. CONCLUSION: Treatment of dumbbell schwannoma is surgical, dilemmas of the optimal surgical approach. When it is thoracally located the posterior approach with hemilaminectomy and costotransversectomy is safe and effective for its removal. PMID- 20017424 TI - [Application of palatal RB obturator in babies with isolated palatal cleft]. AB - BACKGROUND: Isolated cleft of secondary palate has a specific clinical picture due to a wide communication between the mouth and nose cavity. As a consequence of that, babies born with this malformation are often subject to infections of the upper bronchial tubes, middle ear, speech disorders, and certainly the most difficult existential problem they face at the very beginning of their lives, the impossibility of suckling (breast feeding). Such babies have to be fed with gastric probe. The difficulties in their nutrition have often been described in literature, yet a singular attitude toward early orthodontic therapy has not been adopted still. The aim of the paper was to describe a design and application of obturator immediately after the birth of a baby with isolated palatal cleft, and the role in feeding. CASE REPORT: We presented a female neonate, born on 27th December 2007, with a wide fissure in the shape of the letter U over the entire secondary palate. The baby was referred to the Stomatology Clinic due to nutrition impossibility. To avoid feeding with gastric probe, the formation of RB obturator was performed (artificial palate). Hereby, the procedure of obturator making with an explanation of its function is presented. CONCLUSION: The application of RB obturator and the necessary education of parents have a major role in shortening the time of breast feeding and increasing the amount of food intake and, thus, for the normal growth and development of newborn infants with isolated palate cleft. PMID- 20017425 TI - Large saphenous venous graft aneurysm mimicking atypical mediastinal mass. AB - BACKGROUND: Saphenous venous graft (SVG) aneurysm is a very rare but potentially fatal complication of the coronary artery bypass surgery. CASE REPORT: We reported a case of 72-year-old man admitted to hospital because of atypical chest pain related to body motions in horizontal position, especially to the left side. Pain was followed by dispnea, palpitations, fatigue, cough, yellow sputum expectorations, as well as elevated temperature. He had had coronary artery bypass grafting (CABG) surgery with saphenous vein grafts (SVGs) to the left anterior descending artery (LAD) and right coronary artery (RCA) 27 years earlier. Chest X-ray revealed a poor-defined shadow in the region of the right atrium. A transthoracic echocardiogram revealed an atypical tumorous mediastinal mass near the right atrium and right ventricle that seemed partially calcified on transesophaeal echocardiography (TEE). CT scan confirmed an atypical mediastinal mass in contact with the right ventricle that might be a right ventricle aneurysm, pericardial cyst or SVG aneurysm. Coronary angiography was performed subsequently and it revealed a big saphenous venous graft aneurysm originating from the previous venous graft to the RCA. The aneurysm was resected and a new bypass graft was placed. Histopathology confirmed a true aneurysm of the venous graft. CONCLUSION: Although SVG aneurysm is a very rare complication of CABG surgery, patients presenting with atypical hilar or mediastinal mass following CABG should always be evaluated firstly for existence of this cardiosurgical complication. PMID- 20017426 TI - Liver actinomycosis mimicking liver tumour. AB - BACKGROUND: The liver actinomycosis is a rare disease associated with complex differentiation from the liver metastases or hepatocellular carcinoma. CASE REPORT: A 50-year-old immunocompetent female patient was admitted to the Surgical Department in an exhausted condition, with dyspnea, significant weight loss and intermittent fever in the recent two months. Diagnostic procedures that followed, including abdominal ultrasound and computed tomography led us to the diagnosis of metastatic liver disease of unknown etiology with pleural and pericardial effusion. Intraoperatively, the presence of liver pseudotumor without malignancy in the liver was confirmed. Histological examination confirmed the diagnosis of liver actinomycosis. Prolonged treatment with high dose penicillin was performed and all signs and symptoms resolved completely without further problems. The control abdominal ultrasond finding was normal. CONCLUSION: Liver actinomycosis has a nonspecific presentation, often mimicking liver tumor. A timely diagnosis as well as a combined surgical and antibiotic therapy is necessary in the treatment of patients with primary disease and prevention of complications. PMID- 20017427 TI - Use of recombinant factor VIIa in the treatment of massive retroperitoneal bleeding due to severe necrotizing pancreatitis. AB - BACKGROUND: Recently, a growing number of case reports and case series have suggested that the use of recombinant activated factor VII (rFVIIa) may be effective in treatment of patients with non-hemophilic acquired coagulopathy not responding to conventional treatment such as major surgery, major trauma, sepsis, necrotizing pancreatitis and bleeding due to cerebral arteriovenous malformations. CASE REPORT: We presented a septic patient with massive, life threatening bleeding caused by retroperitoneal necrosis, due to severe acute necrotizing pancreatitis. As conservative treatment (blood, plasma, cryoprecipitates and platelet transfusions) failed to induce cessation of bleeding, the patient was urgently operated on. In spite of usual procedures of surgical hemostasis (ligation, suture, thermocauterisation, fibrin glue, temporary tamponade), hemorrhage could not be stopped. The patient manifested the signs of hypothermia and metabolic acidosis and, therefore, the decision was made to use recombinant activated factor VII (Novo Seven). The application of rFVIIa resulted in significant discontinuation of hemorrhage, restoration to normal blood count as well as other relevant coagulation parameters. CONCLUSION: Although application of rFVIIa is still in the initial clinical phase, and the experience is based mainly on uncontrolled series as well as on individual observations, it seems that this drug can be promising, potent and attractive adjunctive prohemostatic agent. This drug may play a beneficial role in the treatment of serious and unresponsive, "nonsurgical", life-threatening bleeding due to severe acute necrotizing pancreatitis. PMID- 20017428 TI - In the light of evolution. PMID- 20017429 TI - [Neuroprotection following cardiac arrest and early post-resuscitation care]. PMID- 20017430 TI - [Is there a place for thromboelastography in cardiac surgery and cardiology?]. PMID- 20017431 TI - [Long-term evaluation of patients with type 1 diabetes mellitus treated with insulin glargine]. AB - AIMS OF THE STUDY: To evaluate long-term effects of treatment with insulin analogue glargine in patients with type 1 diabetes mellitus and to follow up their further course of life. PATIENT SAMPLE AND METHODOLOGY: Retrospective evaluation of 114 patients who, from September 2004, had their basal insulin changed from NPH insulin to insulin glargine. Treatment was changed again in patients in whom a year-long treatment with insulin glargine did not bring improvement in diabetes control. The original sample was divided into 3 groups and the results compared. Compensation of diabetes (HbA1c) after 1, 2 and 3 years and changes to basal and bolus daily insulin dose and body weight were evaluated. RESULTS: The results are presented as median and 25th and 75th percentile. Group A--75 patients (65%) treated for the entire evaluation period with insulin glargine. Initial HbA1c was 7.3 (6.4-8.2)%, 6.9 (6.0-8.4)% after 1 year, 7.1 (5.9 7.9)% after 2 years and 6.6 (5.5-7.7)% after 3 years (p < 0.001). We did not identify any statistically significant changes to total, basal or bolus daily dose of insulin or statistically significant body weight increase over the evaluation period. Group B--19 patients (17%). Switch from insulin glargine to detemir twice daily. Initial HbA1c was 7.3 (6.9-8.5)%, 7.4 (6.8-8.7)% after 1 year of treatment with insulin glargine, 7.7 (7.2-8.1)% before the treatment switch and 7.8 (6.7-8.5)% (NS) after 3 years of treatment. Daily dose of total, basal and bolus insulin did not change and, similarly, no statistically significant change to patients' bodyweight was identified. Group C--17 patients (15%). Switch from insulin glargine to an insulin pump. This group had better initial compensation with HbA1c 6.7(5.7-8.6)%, HbA1c after 1 year was 6.2 (5.6 8.1)%, 7.0 (6.0-7.4)% before the treatment switch and 6.3 (5.2-7.7)% after 3 years of treatment. Total daily insulin dose: 48 (34-60)-38 (25-49) IU/day (NS). Basal daily insulin dose: 17.5 (13-28) IU/day-23 (12-32) IU/day (NS). Bolus daily dose decreased significantly: from 25.5 (21-33) to 15.5 (12-22) IU/day (p < 0.01). Body weight: 76 (71-97) kg-73 (72-99) kg (NS). Only 3% of patients went back to NPH insulin. CONCLUSION: Insulin glargine brings improved control of diabetes. The dose of insulin glargine did not differ from NPH insulin. No statistically significant body weight increase was observed during the evaluation period. PMID- 20017432 TI - [Inflammatory bowel disease--familial and sporadic form]. AB - THE AIM: The aim of this thesis was to elucidate more differences between a familial and sporadic inflammatory bowel disease by comparing certain clinical data. METHODS: We assessed 248 patients with inflammatory bowel disease (IBD) observed in 1994-2004 in the Academic Department of Gastroenterology at the Medical Faculty in Hradec Kralove. To get information about the defined characters we obtained from the questionary and the hospital data. RESULTS: We did not identify any relationship between the onset of the disease and a certain age group, yet males seem to be more prone to familial Crohn's disease. The more frequent familial form of Crohn's disease was the fibro-stenotic one. There were no differences in the onset of the disease. We did not prove the differences in extraintestinal signs, alergy and comorbidities. We did not find any differences in therapy response in relation to the type of nutrition (enteral, parenteral) and the administration of immunosupresive drugs. The biological therapy in sporadic and familial Crohn's disease did not differ either. Surgical intervention was more frequent in Crohn's patients compared to the patients with ulcerative colitis; yet no difference was identified between familial and sporadic cases. Appendectomy carried out before the onset of the disease was later diagnosed as Crohn's disease in more instances than ulcerative colitis. CONCLUSION: We did not prove significant differences comparing certain clinical data in familial and sporadic form of inflammatory bowel disease, yet males seem to be more prone to familial Crohn's disease. Small bowel was involved more often in familial form of Crohn's disease than in sporadic form. PMID- 20017433 TI - [Social, economic and psychological issues as cardiovascular disease risk factors]. AB - The highest mortality in developed countries is, according to the data from our as well as international literature, among people from low socioeconomic backgrounds. No recent data are available for the Czech Republic. We compared the estimated median life expectancy in 2001-2005 with the proportion of people with basic education only and with the proportion of unemployed in all regions within the Czech Republic. We confirmed significant negative correlation--people in regions with low socioeconomic status had lower estimated median life expectancy. CONCLUSION: Despite the decline in the total and cardiovascular mortality in particular in the Czech Republic over the last 20 years, the highest mortality and shortest medium life expectancy continue to persist in the regions with the lowest socioeconomic status as measured by the educational status and unemployment. PMID- 20017434 TI - [Examination of function and structure of respiratory cilia of adult patients suffering from chronic obstructive pulmonary disease (COPD)--comparison of nasal and bronchial mucosa (pilot of CILIARY STUDY)]. AB - INTRODUCTION: Borderline between upper and lower respiratory tract pathology is probably artificial (bronchial asthma). Also inflammation of bronchial mucosa during chronic obstructive pulmonary disease (COPD) is likely combined with inflammatory involvement of nasal mucosa. Ciliary edge of respiratory epithelium is very important part of mucosa layer. AIM: To investigate and compare nasal and bronchial ciliary beat frequency (CBF), degree of nasal and bronchial ciliary dyskinesia, presence of ciliary akinesia and incidence of spinocellular metaplasia in the both mucosa localities among of stable COPD patients (pts). METHOD: Nasal and bronchial mucosa were obtained in the course of bronchoscopy examination of COPD pts in general intravenous anesthesia. Native samples of mucosa tissue were assessed by digital high-speed video microscopy (1,000x magnification). Paired t-test was used to evaluate differences in average frequencies. Significance level was alpha = 0.05. Mode was used to describe "index of dyskinesia", as a measure of association was used K coefficient. MATERIAL: Seventeen COPD pts (6 weeks free of exacerbation) at the age 47-80 (average 64.2 years +/- 9.7) were examined (13 male), average FEV1 61% predic. value (21-81, +/- 15). All patients were active smokers (average 42 pack years +/ 22.8) and all suffered from bronchitic (daily sputum production) phenotype of COPD. RESULTS: We did not find any difference in average ciliary beat frequencies between nose (6.0 Hz +/- 1.3) and bronchus (5.9 +/- 1.3) locality (p = 0.427). We find weak association between nose and bronchus in "ciliary akinesia" (kappa = 0.282) but medium association in "metaplasia" (kappa = 0.485), in index of dyskinesia (kappa = 0.733). CONCLUSION: We did not find in our data any difference in nasal and bronchial ciliary beat frequencies and we found medium association between nasal and bronchial spinocellular metaplasia and index of ciliary dyskinesia. Possible generalization of these results would require further investigation and analysis. PMID- 20017435 TI - [Endoscopic diagnostics and management of pancreatic-biliary disorders in patients after Billroth II gastric resection]. AB - AIMS OF THE STUDY: The aim of this retrospective study was to analyse diagnostic and therapeutic success of endoscopic retrograde cholangiopancreatography (ERCP) in our sample of patients following Billroth II gastric resection, where, due to significantly modified anatomic ratios, this surgery represents a specific and often extremely difficult technical problem when performing ERCP. MATERIALS AND METHODOLOGY: The sample was followed up for 13 years (November 1994-December 2007). The data on 112 patients after Billroth II gastric resection were assessed retrospectively; indications for ERCP included cholestasis in 92 patients, acute biliary pancreatitis in 12 patients, acute cholangitis in 6 patients and suspected bile leak following laparoscopic cholecystectomy (LCE) in 2 patients. RESULTS: Cannulation success during ERCP in the 112 patients following Billroth II gastric resection was 90.2% (i.e. 101 of the 112 patients). Normal ERCP finding was recorded in 4 patients. The remaining 97 patients had pathological results on ERCP (choledocholitiasis was found in 78 patients, malignant biliary stenosis in 14, benign biliary stenosis in 3 a bile leak following LCE in 2). Endoscopic treatment was initiated immediately after diagnostic ERCP in all these 97 patients, the initial step was in all cases endoscopic papillotomy using one of the special papillotomes (diathermy wire). Overall, therapeutic ERCP was completely successful in 83 of the 97 patients (85.6% of 97) in whom the originally endoscopic treatment had been initiated. CONCLUSIONS: ERCP following Billroth II gastric resection is, due to modified post-surgery anatomy, markedly more challenging then the conventional procedure. Availability of a variety of tools as well as, understandably, extensive experience and skill of an endoscopist are prerequisite to ERCP success in these patients. Correctly performed ERCP in patients following Billroth II gastric resection is a highly effective and safe method for diagnostics and, in particular, treatment of pancreatic-biliary diseases, in which similar success as under standard anatomic conditions can be achieved. PMID- 20017436 TI - [Neuroendocrine tumours of the appendix]. AB - According to WHO, neuroendocrine tumors of the appendix (appendiceal carcinoids) are defined as 1. well-differentiated endocrine tumors with benign or uncertain behavior, 2. well-differentiated endocrine carcinoma and 3. goblet cell carcinoma. These tumors are usually diagnosed incidentally during appendectomy. Carcinoid syndrome is rare in appendiceal carcinoid. Tumor size greater than 2 cm is the most important parameter for prognosis. Most patients are cured by appendectomy (appendiceal tumors < or = 2 cm), tumors with a diameter > 2 cm should be managed by right hemicolectomy. PMID- 20017437 TI - [Goblet cell carcinoid of the appendix]. AB - Appendiceal goblet cell carcinoids are mixed tumors including neuroendocrine cells and intestinal type of goblet cells. Compared to typical carcinoids, goblet cell carcinoids are malignant tumors with degree of malignity differing from case to case. Carcinoid syndrom is not present. There is poor prognosis in larger tumors. Appendectomy is a sufficient therapeutical approach in small tumors, hemicolectomy should be performed in larger ones. There is limited experience with chemotherapy in metastasizing tumors and sporadic with somatostatin analogues. PMID- 20017438 TI - [Current cooling methods for induction of mild hypothermia in cardiac arrest survivors]. AB - Induction of mild therapeutic hypothermia early after return of spontaneous circulation improves prognosis of cardiac arrest survivors. Rapid cooling of the patients and correct maintainance of the target therapeutic temperature followed by controlled slow rewarming can be achieved by several noninvasive and invasive methods of various efficacy. Elementary and the most frequently used methods are surface cooling via ice-packs and rapid intravenous administration of cold crystaloids. Mattress cooling systems and facilities for endovascular cathether cooling are more sophisticated, manageable and ensure more precise titration of therapeutic temperature. Cooling caps and helmets leading to selective head cooling can be used as the complementary techniques. Several other methods are too instrumentation-intensive, too invasive or investigated in animal experiments only. Anyway, near future may bring a rapid development of new effective and safe cooling systems. PMID- 20017439 TI - [Is thromboelastography able to influence management of patients in cardiology and cardiosurgery?]. AB - Thromboelastography (TEG) is perceived as a non-specific test of hemostasis. It evaluates the global hemocoagulation, however by some modifications could be its diagnostic potential greater. With new inovations isTEG except more precise analysis of participation of blood components on final coagulation (for example rotational TEG) capable also to assess the antiplatelet drug nonresponsiveness (for example platelet mapping system). It gives more accurate information about prothrombotic conditions than routine laboratory techniques in state of ventricular assist device implantation. PMID- 20017440 TI - [High sensitivity cardiac troponin assays 2009: clinical potential, current practice and benefits, the future]. AB - At present, determination of cardiac troponins (cTn) is the biomarker method of choice for diagnostics and risk stratification in patients with a myocardial injury. Past clinical practice had provided sound evidence that low cTn concentrations, measured with unacceptable imprecision by the currently used methods, hold important clinical, diagnostic and stratification potential. The new generation cTn assays, so called high-sensitivity assays, enable determination of very low cTn concentrations with satisfactory analytical precision and open the way to early identification of small but often prognostically important myocardial damage. Introduction of high-sensitivity cTn assays in practice is, however, associated with some difficulties: their superior diagnostic sensitivity to identify small injuries to myocardium is often linked to lower specificity, higher incidence of elevated cTn concentrations is frequently associated with less obvious clinical symptomatology (overdiagnosis), resulting in greater demand for further patient assessment (overcrowding), repeated analyses and trend monitoring of cTn fluctuation. These initial difficulties cannot lessen the by now indisputable, established benefit of high sensitivity cTn assays that we briefly describe in the present paper. PMID- 20017441 TI - [The results of the RE-lY study promise more effective, safer and easier prevention of embolic complications in patients with non-valvular atrial fibrillation]. AB - The RE-LY study compared dabigatran in the dose of 150 mg and 110 mg twice daily, without laboratory monitoring, with the conventional treatment with warfarin dosed according to INR in 18,113 patients with non-valvular atrial fibrillation and high risk of embolisation. The incidence of cerebrovascular events and systemic embolisation was 1.69% per year in the warfarin group, compared to 1.53% per year in the 110 mg dabigatran group (relative risk 0.91; 95% CI 0.74-1.11; p < 0.001 for non-inferiority) and 1.11% per year in the 150 mg dabigatran (relative risk 0.66; 95% CI 0.53-0.82; p < 0.001 for superiority). Major bleeding occurred in 3.36% of patients per year in the warfarin group, compared to 2.71% of patients per year in the 110 mg dabigatran group (p = 0.003) and 3.11% of patients per year in the 150 mg dabigatran group (p = 0.31). Cerebral haemorrhagic events occurred in 0.38% of patients on warfarin per year, compared to 0.12% per year in the 110 mg dabigatran group (p < 0.001) and 0.10% per year in the 150 mg dabigatran group (p < 0.001). Mortality was 4.13% per year in the warfarin group, compared to 3.75% per year in patients on 110 mg dabigatran (p = 0.13) and 3.64% per year in patients on 150mg dabigatran (p = 0.051). In conclusion, administration of dabigatran to patients with atrial fibrillation in the dose of 110 mg in the RE-LY study was associated with the same incidence of cerebrovascular events and systemic embolisations as with warfarin, while there was lower incidence of major bleeding complications. Dabigatran in the dose of 150mg compared to warfarin led to reduction in the incidence of cerebral events and systemic embolisations with the same incidence of haemorrhagic complications. PMID- 20017442 TI - [Complete remission of nephrotic syndrome and improvement of renal function in a patient with light chain deposition disease following high dose chemotherapy with transplantation of autologous haematopoietic stem cells. A case study and review of literature]. AB - Light chain deposition disease (LCDD) damages most frequently kidneys, and less frequently other organs. The incidence of LCDD is lower than the incidence of AL amyloidosis. Symmetric swelling of both legs was the first sign of nephrotic syndrome with renal insufficiency in our female patient. Renal biopsy specimen revealed the diagnosis of LCDD. Bone marrow biopsy contained 40% of plasma cells. Bone survey showed no osteolytic changes. These findings confirmed the diagnosis of multiple myeloma (MM) Durie Salmon stage IB with LCDD. The patient was initially treated with 4 cycles of VAD (vinkristine, adriamycine, dexamethasone) chemotherapy with no response. Followed collection of peripheral haematopoietic stem cells and later high dose chemotherapy with reduced dose of melphalan 140 mg/sqm and autologous peripheral haematopoietic stem cells transplantation. Melphalan dose was reduced because of renal insufficiency (serum creatinine 290 micromol/l) before application of conditioning regimen. High dose therapy was complicated by with deterioration of renal function, creatinine increased to 600 micromol/l. Worsening of renal function was most likely caused by nephrotoxicity of melphalan in nephrotic syndrome. This has been previously described in patients with AL-amyloidosis, and nephrotic syndrome who were treated with high dose melphalan. This phenomenon was entitled "post conditioning renal insufficiency". Hypoalbuminemia hypoproteinemia and reduced intravascular volume and renal damage caused by amyloid deposits as well as probably, amorphous non amyloid deposits of monoclonal immunoglobulin are likely to have contributed to nephrotoxicity of the high dose of melphalan. However, worsening of renal insufficiency was facilitated by the mucositis-associated sepsis. Follow-up examination one month after high dose chemotherapy showed complete remission, that was confirmed by further examinations. In the course of the first year after high dose chemotherapy renal function gradually improved and nephrotic syndrome completely disappeared (complete kidney remission). Proteinuria declined to 2-3 g/24 hours and glomerular filtration slowly improved. Three years after high dose chemotherapy the patient is still in complete remission of multiple myeloma and free of nephrotic syndrome, with slightly increased creatinine (160 micromol/l) that, nevertheless, has had an improving tendency over last 3 years. The present case study illustrates accomplishment of complete haematological remission with high dose chemotherapy followed by autologous haematopoietic stem cells transplantation despite complete resistance of the disease to the standard chemotherapy VAD in a patient with MM and LCDD. We draw the reader's attention to the possibility of nephrotoxic effects of high dose melphalan (post conditioning renal insufficiency) in patients with nephrotic syndrome caused by light chain deposits as AL-amyloid or amorphous light chains deposits (LCDD)and we document the importance of plasma free light chain detection. PMID- 20017443 TI - [Recurrent unsustained ventricular tachycardia in pregnancy]. AB - The incidence of arrhythmias in pregnancy is more frequent. Arrhythmias can develop in women withstructural heart disease as well as in healthy population. Palpitations, dizziness, vertigo or syncope are the most common symptoms of arrhythmias. Rarely life threatening arrhythmias requiring acute therapy occur. The treatment of the pregnant patient with cardiac arrhythmias requires important modifications of the standard practice. We describe pregnant patient in 33rd week of pregnancy with new onset of recurrent unsustained ventricular tachycardia, which did not regress after the pharmacological therapy. Patient was indicated to cesarean section, after which the arrhythmias completely regressed. PMID- 20017444 TI - [Repeated local thrombolysis in a patient with axillary and subclavian vein injury and thrombosis]. AB - Deep vein thrombosis (DVT) is a disorder with blood clot (thrombus) formation in deep veins. DVT of upper extremities (UE) is rare but serious, bearing in mind its possible complications, disease. UE DVT treatment involves subcutaneous injections of low molecular weight heparins and subsequent switch to oral anticoagulation or thrombolysis; thrombolysis by intravenous administration of a thrombolytic agent is used only if indicated (subclavian vein thrombosis). A case of a professional sportswoman is presented, who was treated for venous thrombosis of subclavian and axillary veins using local thrombolysis. Thrombosis emerged suddenly after a basketball match, during which oedema of the right arm occurred subsequent to the patient being hit with the ball to the armpit area. Venipuncture as well as all catheter repositionings were performed under the duplex ultrasound (DUS) surveillance. Proximal brachial vein was punctured and four French catheters were used to enter subclavian vein. For thrombolysis, 0.2 0.6 mg/hour alteplase (rt-PA) was used. Injury to axillary vein tunica intima was evident the following day after the subclavian and axillary thrombosis was dissolved completely, and thrombosis in the same area recurred immediately after stopping the thrombolytic infusion. Therefore, administration of a thrombolytic agent into the same area was re-introduced and acetylsalicylic acid was added to the medication. This combined therapy resulted in recanalization of the arm as well as the axillary and subclavian areas. Thrombolysis was stopped on the 3rd day of treatment. The method can be used to manage deep vein thromboses of the upper arms even without X-ray control. Local thrombolysis of deep vein thrombosis with ultrasound surveillance is, in indicated patients, a safe method even if a vein injury occurs. PMID- 20017445 TI - [The results of patients with essentials thrombocythemia and other myeloproliferation-related thrombocythemia--a report of patients treated with Thromboreductin]. AB - The registry of patients treated with Thromboreductin (anagrelide) in the Czech Republic contains data concerning patients that have been treated using this drug since 2004. As of June 2009, the total number of patients was 549. The current analysis focused mainly on evaluation of anagrelide dosage needed to achieve a complete response in high-risk patients: reduction in platelet count to below 400 x 10(9)/l, which was also considered as reaching the therapeutic goal. The outcomes of the registry confirm that although anagrelide (Thromboreductin) is a very effective platelet-reducing agent, the administration of which is related to a low incidence of adverse effects and complications, the therapeutic goal is not achieved in all cases and or despite a quick treatment response, the therapeutic goal is achieved more slowly. PMID- 20017446 TI - Warm liver ischemia in experiment and lysosomal markers. AB - BACKGROUND: The aim of the study is to perform a morphological analysis of certain lysosomal enzymes and parenchymal alterations during warm ischemia in the pig liver. METHODS: Standard hepatectomies were performed in a set of 24 pigs. Intra-operative intravenous (portal vein) Pentoxiphylline and hydroxyl radical scavenger Stobadine was administered. Tissue specimens were removed from the margo acutum in 10 minutes interval. RESULT: In normal pig liver, the acid phosphatase (ACP) activity is in not numerous Kupffer cells and on the biliary pole of hepatocytes, diffusely in the whole parenchyma. One hour after the beginning of warm ischemia, there was an increase in ACP activity in the cytoplasm of hepatocytes. The activity in Kupffer cells could not be detected. Lactate dehydrogenase (LDH) is localized exclusively in the cytoplasmic matrix of liver cells, so only cytoplasmic enzymes leak into the blood plasma. LDH activity has remained low in areas around portal and central veins. CONCLUSION: Morphological findings of enzyme activities showed that zone 2 and 3 of the liver lobule are essential for the organ survival and signs of diffusion of lysosomal enzymes into the cytoplasm of hepatocytes indicate one of the possible explanations for the findings after liver reperfusion. The study showed that intravenous administration of Pentoxiphylline and Stobadine protects the liver from warm ischemia injury (Tab. 2, Fig. 2, Ref. 37). Full Text (Free, PDF) www.bmj.sk. PMID- 20017447 TI - Arginase in the airways hyperreactivity. AB - BACKGROUND: The interest of arginase action is increasing because limitation of L arginine bioavailability by arginase for NO synthesis via constitutive NOS can contribute to airway hyperreactivity. OBJECTIVES: We investigated the effect of intervention in the arginase activity in guinea pig model of experimental ovalbumin-induced airway hyperreactivity. METHODS: We analysed the response of tracheal and lung tissue smooth muscle strips to histamine or acetylcholine after in vitro administration of arginase in a dose of 75 UI or after administration of the non-selective inhibitor of arginase N(omega)-hydroxy-L-arginine (NOHA) in a dose of 5 and 10 micromol. We used as well as the incubation of strips with the precursor of NO synthesis L-arginine in a dose of 10(-4) mol/l together with NOHA. RESULTS: We did not find any significant differences in the reactivity of tracheal and lung tissue smooth muscle if we applied arginase in a dose of 75 UI in vitro. NOHA in a dose of 5 a 10 micromol induced the decrease of tracheal and lung tissue smooth muscle reactivity overall. The decrease of the contraction amplitude was dose-dependent. The supplementation of NO synthesis precursor L arginine in a dose of 10(-4) mol/l together with NOHA intensified the decrease of the airways reactivity induced by an arginase inhibition. CONCLUSION: The results suggest that arginase is involved in the control of airways bronchomotoric tone and therefore modulation of arginase activity could be a useful tool for airway smooth muscle tone control in clinical conditions (Fig. 7, Ref. 33). Full Text (Free, PDF) www.bmj.sk. PMID- 20017448 TI - Fecal calprotectin as a promising marker of inflammatory diseases. AB - BACKGROUND: Calprotectin is a calcium binding protein present predominantly in neutrophils with antimicrobial and antiprolipherative activities. Calprotectin concentration is higher in feces than in plasma and significantly increased levels of fecal calprotectin (FC) were found in patients with bowel inflammation disease (IBD). METHODS: Nineteen out-patients with IBD, comprising 14 Crohn's disease (CD) individuals and 5 ulcerative colitis (UC) patients, and 5 healthy volunteers were investigated. Fecal calprotectin was analyzed by ELISA. RESULTS: We found that patients with IBD had significantly higher concentration of FC than in healthy children (FC median 1076.7 vs 19.5 microg/g of stool, p = 0.0053). We determined higher level of FC in patients with CD than in UC (1132.4 vs 490.98 microg/g of stool), but not statistically significant. CONCLUSION: It has been proved that FC represents a surrogate marker of neutrophils influx into the bowel lumen; hence it can be regarded as a simple and non-invasive marker of intestinal inflammation (Tab. 2, Fig. 1, Ref. 38). Full Text (Free, PDF) www.bmj.sk. PMID- 20017449 TI - Pulmonary arterial hypertension--contemporary management strategy. AB - Pulmonary arterial hypertension (PAH) is a severe chronic disorder of pulmonary arteries with progressive precapillary pulmonary hypertension, characterized by poor life quality and very poor prognosis. Unless treated, it causes death within 2-3 years from diagnosis. PAH affects mainly younger women. The treatment of PAH should not only be symptomatic, but also directed towards the improvement in patient's survival and quality of life. Many novel drugs putting together so called specific PAH therapy (endothelin receptor antagonists, prostanoids, phosphodiesterase--5 inhibitors) were tested in randomized trials. PAH management requires a highly individualized approach, state of the art knowledge and adequate experience. Patients therefore should be referred to specialized PAH centers providing both complete diagnosis and therapy. In our region a close co operation between Czech and Slovak PAH centers has also proved to be profitable. Data sources. Literature retrieval was accessed through MEDLINE using the terms pulmonary hypertension, PAH, diagnosis, treatment. Reference citations from publications identified were reviewed (Ref. 47). Full Text (Free, PDF) www.bmj.sk. PMID- 20017450 TI - Cochlear implantation in malformed inner ear. AB - OBJECTIVES: Authors evaluate diagnosis, surgery and results in the implantees with malformed inner ear. METHODS: Four children with the inner ear malformations were included in this series (one child with common cavity, two children with narrow internal auditory meatus and one child with lateral semicircular canal aplasia). CT and MRI were used for the imaging of inner ear malformations. Different types of implants (Medel Combi 40+, Nucleus Freedom) with different types of electrodes were used. The average time of being the cochlear implant user in the time of evaluation was 3 years. RESULTS: Pure tone audiometry (PTA), Category of Auditory Performances (CAP), Evaluation of spontaneous speech and Evaluation of speech understanding were used to evaluate of the CI outcome. CONCLUSIONS: Malformation of inner ear with the developed cochlear nerve nowadays is the indication for CI. Councelling with parents must be open and detailed to inform them on realistic expectations and different ways of deaf child education (Tab. 4, Fig. 4, Ref. 13). Full Text (Free, PDF) www.bmj.sk. PMID- 20017451 TI - New approach to probability estimate of femoral neck fracture by fall (Slovak regression model). AB - PATIENTS AND METHODS: 3,216 Slovak women with primary or secondary osteoporosis or osteopenia, aged 20-89 years, were examined with the bone densitometer DXA (dual energy X-ray absorptiometry, GE, Prodigy - Primo), x = 58.9, 95% C.I. (58.42; 59.38). The values of the following variables for each patient were measured: FSI (femur strength index), T-score total hip left, alpha angle - left, theta angle - left, HAL (hip axis length) left, BMI (body mass index) was calculated from the height and weight of the patients. RESULTS: Regression model determined the following order of independent variables according to the intensity of their influence upon the occurrence of values of dependent FSI variable: 1. BMI, 2. theta angle, 3. T-score total hip, 4. alpha angle, 5. HAL. The regression model equation, calculated from the variables monitored in the study, enables a doctor in praxis to determine the probability magnitude (absolute risk) for the occurrence of pathological value of FSI (FSI < 1) in the femoral neck area, i. e., allows for probability estimate of a femoral neck fracture by fall for Slovak women. CONCLUSION: 1. The Slovak regression model differs from regression models, published until now, in chosen independent variables and a dependent variable, belonging to biomechanical variables, characterising the bone quality. 2. The Slovak regression model excludes the inaccuracies of other models, which are not able to define precisely the current and past clinical condition of tested patients (e.g., to define the length and dose of exposure to risk factors). 3. The Slovak regression model opens the way to a new method of estimating the probability (absolute risk) or the odds for a femoral neck fracture by fall, based upon the bone quality determination. 4. It is assumed that the development will proceed by improving the methods enabling to measure the bone quality, determining the probability of fracture by fall (Tab. 6, Fig. 3, Ref. 22). Full Text (Free, PDF) www.bmj.sk. PMID- 20017452 TI - Surgical treatment of pulmonary metastases in germ-cell testicular cancer patients--long-term results. AB - OBJECTIVES: The aim of this prospective clinical study is to evaluate our 20-year experience with surgical treatment of pulmonary metastases in germ-cell testicular cancer and discuss the optimal therapeutical approach. BACKGROUND: Authors presents 63 patients who were surgically treated in the period from 1988 to 2008 for pulmonary metastases in germ cell testicular cancer. All patients were allocated for surgical treatment after undergoing cisplatin-based combination chemotherapy. METHODS: In a group of 63 patients, 57 thoracotomias and 27 sternotomias were performed. In pulmonary procedures small atypical resections prevailed--84%. Postoperative mortality was 1.2%. Decision for follow up treatment was made on basis of histological investigation of metastases. In some cases when viable tumor tissue was found (16% patients), further chemotherapy was administered. The rest of patients were monitored intensively. RESULTS: In a long-term follow-up, remission was achieved in almost 60% of patients. CONCLUSION: We suggest that lung metastasectomy as the secondary treatment in generalized germ cell testicular cancer should be fully warranted (Fig. 3, Ref. 16). Full Text (Free, PDF) www.bmj.sk. PMID- 20017453 TI - Procalcitonin, neopterin and C-reactive protein in diagnostics of intrauterine infection and preterm delivery. AB - OBJECTIVE: The purpose of this study was to find out whether Procalcitoni, Neopterin and C-reactive protein are sensitive and specific markers of intrauterine infection. METHODS: We evaluated 155 patients from 26. to 41. week of pregnancy at the time of delivery. We measured serum concentrations of procalcitonin (PCT), neopterin and C-reactive protein (CRP) from mother's blood sample at the beginning of delivery and from umbilical cord blood after delivery. RESULTS: In first group occurred in higher percentage (27.41%) preterm delivery (26.-37. week of pregnancy), chorioamnionitis confirmed by histological examination (16.12%) and preterm premature rupture of membranes (24.19%). In this group occured perinatal infection of newborn in 61.29%. In the second group preterm delivery (6.31%) and perinatal infection of newborn (7.36%) occured in lower percentage. CONCLUSION: The results suggest that the simultaneous measurement of CRP, PCT and NPT in mother's blood sample before delivery and umbilical cord blood may provide an accurate early diagnosis of infection and then preterm delivery (Tab. 1, Fig. 3, Ref. 18). Full Text (Free, PDF) www.bmj.sk. PMID- 20017454 TI - Increase in the circulating level of hepatocyte growth factor in pancreatic cancer patients. AB - OBJECTIVE: Hepatocyte growth factor (HGF) has been reported the cause of many biological events, including cell proliferation, invasiveness, morphogenesis, and angiogenesis. Elevated HGF content in tumor tissue was reported to predict a more aggressive biology in breast and gastric cancer patients. MATERIALS AND METHODS: Eighty patients with invasive pancreatic cancer investigated. Venous blood samples were collected before the surgery. Sera were obtained by centrifugation and stored at -70 degrees C until assayed. The control group created from healthy individuals. Serum concentrations of soluble HGF were measured by the quantitative sandwich enzyme immunoassay technique. RESULTS: The mean value of serum soluble HGF in patients with invasive pancreatic cancer was 497.2 +/- 53.8 pg/ml and that of control group was 53.6 +/- 7.5 pg/ml and the difference was significant (p < 0.001). CONCLUSION: The serum levels of soluble HGF might reflect the severity of invasive pancreatic cancer and deserve further evaluation (Tab. 2, Ref. 19). Full Text (Free, PDF) www.bmj.sk. PMID- 20017455 TI - Systemic malignancies presenting as primary osteolytic lesion. AB - The tumor formation may be the earliest manifestation preceeding other symptoms, signs and bone marrow evidence of systemic malignancy - leukemia/lymphoma. Here we present three cases of systemic malignancy in which bone lesions were the first manifested signs of the disease. All three cases were thought to be orthopedic cases and had been treated as so without genuing improvement. We would like to draw an attention to children who present with multifocal musculoskeletal pain and the importance of whole-body scaning. We describe interesting cases of diffuse large cell lymphoma and leukemia that initially presented as primary osteolytic bone lesion and discuss the differential diagnosis, literature review of non-Hodgkin's lymphoma arising in bone as the primary site (Tab. 1, Fig. 3, Ref. 18). Full Text (Free, PDF) www.bmj.sk. PMID- 20017456 TI - Monosodium glutamate induces apoptosis in naive and memory human B cells. AB - The aim of this study was to establish the existence of mGluR7 in normal B lymphocytes and analyse the effect of monosodium glutamate (MSG) on B cell apoptosis in vitro. B cells were purified by magnetic cell sorting using anti CD19-coupled magnetic beads. Cells (10(6)/ml) were cultured with increasing MSG concentrations (1-100 mM). Detection of apoptosis by flow cytometry was performed using the Annexin V-FITC/Propidium iodide (PI) apoptosis detection kit. Naive and memory B cell population were identified by CD27 staining. Expression of GluRs was determined using PCR. Exposure to increasing MSG concentrations displayed dose dependent effect on B cell viability altogether, ranging from 35% with 100 mM up to 80% with 1 mM MSG. Moreover, the number of late apoptotic cells as well as necrotic cells was dose dependant. Both CD27- as well as CD27+ B cells were affected by MSG. Basal expression of GluRs7 was detected in unstimulated B cells. Glutamate induced apoptosis can be seen in memory as well as naive B cell population and is probably mediated through mGluR7, whose expression in B cells we also confirmed. Our study suggests a new possible mechanism of crosstalk between the nervous and the immune system through glutamate as a potential key mediator (Fig. 4, Ref. 27). Full Text (Free, PDF) www.bmj.sk. PMID- 20017457 TI - Serum and synovial fluid concentrations of CCL2 (MCP-1) chemokine in patients suffering rheumatoid arthritis and osteoarthritis reflect disease activity. AB - OBJECTIVE: To determine serum and synovial fluid (SF) concentrations of monocyte chemoattractant protein-1 (MCP-1) or CCL2 chemokine, in patients suffering (RA) and osteoarthritis (OA) and to correlate the values to disease activity, and other patient- and disease-related parameters. METHODS: The CCL-2/MCP-1 chemokine (CK) was measured in serum and SF of 30 RA and 15 OA patients using specific and very sensitive ELISA assay. RESULTS: The CCL2/MCP-1 CK was found in increased amounts in SF compared to serum (p < 0.001) and in RA compared to OA patients (p < 0.001). The values were significantly greater in RA patients with more active disease. Greater mean SF concentrations were observed in older RA patients, in patients with longer duration of RA disease and in those who had been treated with methotrexate. Also positive correlation was found between RA SF concentrations and SF leukocyte numbers (r = 0.497, p < 0.05). CONCLUSIONS: The SF and serum CCL2/MCP-1 concentrations are significantly greater in RA than in OA and in hda-RA than in mda-RA; increased SF over serum concentrations suggest that CCL2/MCP-1 is mainly produced locally by activated cells where it may exacerbate and sustain inflammation by attracting proinflammatory leukocytes, predominantly monocytes (Tab. 1, Fig. 2, Ref. 50). Full Text (Free, PDF) www.bmj.sk. PMID- 20017458 TI - Renal biopsy in children with steroid-dependent nephrotic syndrome. AB - BACKGROUND: There is lack of consensus on the necessity of renal biopsy in children with steroid-dependent nephrotic syndrome (SDNS) prior to cytotoxic therapy. OBJECTIVES: To retrospectively evaluate: (a) the benefit of renal biopsy (RB) prior to cyclophosphamide therapy; (b) relationship between histopathologic results of RB samples and clinical course in children with SDNS. PATIENTS, MATERIALS AND METHODS: RB was performed in 18 SDNS patients (11 boys and 7 girls). The mean age of the children at the time of nephrotic syndrome (NS) diagnosis was 6.4 +/- 3.9 years and 9.7 +/- 4.3 years at the time of RB. Following the RB, all children received prednisone and cyclophosphamide treatment for 12 weeks. Duration of remission and relapse rate was recorded. RESULTS: The histologic evaluation revealed minimal change disease (MCD; n = 14) and IgM nephropathy (n = 4). These results didn't affect the ongoing therapy. MCD patients had longer remission compared to IgM nephropathy (3.2 +/- 1.5 vs 1.7 +/- 0.8 years; p = 0.05). Relapse rate did not differ significantly between MCD and IgM nephropathy (p = 0.22). The duration of remission was inversely correlated to relapse rate after the treatment (r = -0.66, p = 0.01). CONCLUSION: We suggest that RB prior to cyclophosphamide therapy is not necessary in patients with SDNS (Tab. 2, Ref. 14). Full Text (Free, PDF) www.bmj.sk. PMID- 20017459 TI - Prognostic value of anti-cyclic citrullinated peptide antibodies and rheumatoid factor in patients with rheumatoid arthritis. AB - OBJECTIVES: To determine the predictive value for radiological damage of anti cyclic citrullinated peptide antibodies (anti-CCP) and rheumatoid factor (RF) in patients with rheumatoid arthritis (RA). METHODS: Ninety patients with RA were enrolled in this study. All patients had symptom duration of at least one year. Anti-CCP and IgM-RF were evaluated with enzyme linked immunosorbent assay and nephelometry methods, respectively. Radiological damage was assessed by Larsen score. RESULTS: In forward stepwise logistic regression analysis, anti-CCP positivity and RF positivity were seen as significant independent predictors of the radiological outcomes (p = 0.01, p < 0.05, respectively). The combination of these antibodies had the highest risk for erosive joint damage (odds ratio = 25.71; 95% confidence interval, 4.7 to 140.13; p = 0.001). CONCLUSION: Our results suggest that the combined use of RF and anti-CCP has greater predictive value for erosive RA than anti-CCP or RF alone, and may facilitate to make a decision about the individual treatment in RA (Tab. 4, Ref. 37). Full Text (Free, PDF) www.bmj.sk. PMID- 20017460 TI - Screening for depression in survivors of metastatic ovarian cancer in a programme of palliative cancer care. AB - BACKGROUND: Depression is seen in many cancer patients. It occurs in approximately 25% of palliative care patients. MATERIALS AND METHODS: This study was local, prospective and cross-sectional. It was carried at Department of Clinical Oncology and Radiation Therapy of Charles University Hospital and Faculty of Medicine in Hradec Kralove, Czech Republic. RESULTS: The incidence of depression was 83.3% (25 of all 30 survivors). The relevance of depression is characterized: severely depressed was proved in 9 of all 30 survivors, the moderately depressed in 5 of all 30 survivors, the mildly depressed in 11 of all 30 survivors and normal range in 5 of all 30 survivors. The statistical evaluation not presents statistically significant dependence of ZSRDS on smoking abuse, marital status, age, number of associated diseases and type of palliative cancer care. CONCLUSION: The results of the pilot depression study showed that subsist clear association between metastatic ovarian cancer and depression (Fig. 2, Ref. 30). Full Text (Free, PDF) www.bmj.sk. PMID- 20017461 TI - The physiological conception of thinking based on the results of psychostereotactic operations. AB - The authors have gained their experience in psychostereotactic operations during the past 50 years. The operations were indicated in psychiatric patients on the basis of their clinical pictures. The results gained in operations of the hypothalamus, limbic system and neocortical structures of the brain were logically analysed and served as the basis for physiological conception of the function of the nervous system in the area of thinking. The elemental thinking of the hypothalamus resides in relatively simple memory that contains standard values of the internal environment of the body. The immense memory on the highest neocortical level of the human brain grew to become an independent degree of development independent of the objective reality. The entrance of physiological thinking into psychology enables to clarify the aspects of human psychology that are still liable to subjective view, such as emotions, motivation or consciousnesses (Fig. 1, Ref. 9). Full Text (Free, PDF) www.bmj.sk. PMID- 20017462 TI - Statins: the drugs for the 21st century? AB - This review summarizes the current evidence on beneficial effect of statins on the atherogenesis and the preventive potential of coenzyme Q 10 to avoid statin induced myopathy. Statins, originally discovered in molds are the most effective medication to decrease the low-density cholesterol (LDL-C) which is the key participant in atherogenesis. Statins accomplish this by inhibiting the hydroxymethyl glutaryl coenzyme A reductase, an enzyme that is rate limiting for cholesterol biosynthesis. Inhibition of HMG CoA reductase stimulates in the liver the LDL receptors. The result is an increased clearance of LDL particles from the bloodstream and by this mechanism statins have proved to be highly effective in reducing the cardiovascular risk, as amply documented in clinical studies confirming inhibition of atherosclerosis. Additional beneficial effect of statins occurs by modulation of endothelial functions and by diminishing vessel wall inflammation. Statins reduce the level of the C-reactive protein, a strong predictor of adverse cardiovascular events. Beneficial effects of statins on endothelial dysfunction and synthesis of nitric oxide NO were described. Different statin preparation might differ in their antioxidant effects (Fig. 2, Ref. 46). Full Text (Free, PDF) www.bmj.sk. PMID- 20017463 TI - The use of platelet rich plasma with guided tissue regeneration in defects caused by periodontal diseases. AB - The goal of periodontal treatment in not only the stabilization of disease but also the regeneration of the destructed tissue. In the past few years various procedures have been created to achieve this. The guided tissue regeneration is a surgical procedure developed on the basis of experimental studies. It enables the creation of periodontal tissues affected by periodontitis, the so called reattachment. It stands for formation of new attachment--meaning the regeneration of cementum, alveolar bone and periodontal ligament. This surgical procedure of the treatment of periodontitis is based on the principle of exclusion of the epithelium and also the gingival connective tissue from the root surface so the precursor cells (desmodontal cells) can occupy the defect and pursue their differentiation. Periodontal ligament containing cells with regenerative potential are the exclusive ones to have the ability to regenerate structures affected by periodontitis. The use of growth factors offer new aspects to the therapy (Fig. 7, Ref. 11). Full Text (Free, PDF) www.bmj.sk. PMID- 20017464 TI - Multiple malignity in patients with bronchogenic carcinoma. 3 case reports. PMID- 20017466 TI - Lattice dynamics and thermodynamics of molybdenum from first-principles calculations. AB - We calculated the phase transition, elastic constants, full phonon dispersion curves, and thermal properties of molybdenum (Mo) for a wide range of pressures using density functional theory. Mo is stable in the body-centered-cubic (bcc) structure up to 703 +/- 19 GPa and then transforms to the face-centered close packed (fcc) structure at zero temperature. Under high temperature and pressure, the fcc phase of Mo is more stable than the bcc phase. The calculated phonon dispersion curves accord excellently with experiments. Under pressure, we captured a large softening along H-P in the TA branches. When the volume is compressed to 7.69 A(3), the frequencies along H-P in the TA branches soften to imaginary frequencies, indicating a structural instability. When the pressure increases, the phonon calculations on the fcc Mo predict the stability by promoting the frequencies along Gamma to X and Gamma to L symmetry lines from imaginary to real. The thermal equation of state was also investigated. From the thermal expansion coefficient and the heat capacity, we found that the quasiharmonic approximation was valid only up to about melting point at zero pressure. However, under pressure, the validity can be extended to a much higher temperature. PMID- 20017465 TI - Hydrogen bond lifetimes and clustering of methanol in carbon tetrachloride solutions. AB - NMR experiments and ab initio calculations suggest that methanol forms small cyclic hydrogen bond clusters (4-6 molecules) in the condensed phase. In contrast, molecular dynamics simulations have indicated that methanol will form large branched chains that extend to include hundreds of molecules. In this paper, we performed a series of simulations examining the structure and dynamics of methanol/carbon tetrachloride mixtures. We show that two simulation models are capable of reproducing the trends in the experimental NMR data despite the fact that they indicate that the structure of the liquid is dominated by large branched chains. We hypothesize that the experimental results can be described by variations in the hydrogen bond lifetime with methanol concentration. PMID- 20017467 TI - Theoretical and experimental considerations on the enthalpic relaxation of organic glasses using differential scanning calorimetry. AB - The enthalpy relaxation of amorphous salicin, used as model organic glass of pharmaceutical relevance, was investigated using a combination of DSC measurements and theoretical simulations. The combined approach makes it possible to discern between the effect of the glass forming properties of the material and the effects of the thermal history and experimental conditions. The approach also facilitates an unambiguous definition of the time scale of the experiment, such that objective comparison among relaxation time and glass transition temperature values can be made. The simulation provides accurate predictions of the DSC profiles obtained under a wide variety of experimental conditions. The effects of annealing time and the heating/cooling rate on the enthalpy recovery were explained by tracking the evolution of relaxation times as a function of temperature and time. The combined experimental and simulation approach also makes it possible to systematically explore the effect of specific glass forming properties, such as fragility and nonexponentiality, on the relaxation and associated thermal behavior of molecular organic glasses of pharmaceutical interest. To fully characterize these materials, it is necessary to go beyond the onset T(g) and include the early stages of the glass transition. PMID- 20017468 TI - Proton phototransfers in doubly hydrogen bonded dimers: the photophysics of 6,7,8,9-tetrahydro-5H-pyrido[2,3-b]indole dimers. AB - A photophysical study of 6,7,8,9-tetrahydro-5H-pyrido[2,3-b]indole dissolved in 2 methylbutane at decreasing temperatures from 293 to 93 K revealed the presence of two different fluorescence emissions due to doubly hydrogen-bonded dimers of the monomer species. One of the emissions is assigned to the normal tautomer form of the dimer of C(2) symmetry, and the other is mostly assigned to the most stable dimer, which is nonsymmetric (C(1)) and exhibits single-proton transfer upon electronic excitation. As shown here, when the doubly hydrogen-bonded 7-azaindole dimer loses its centrosymmetry, by fusing a six-membered ring to the molecular structure, its first excited electronic state does not undergo a double-proton transfer but rather a single proton transfer, which, in a fluid medium, causes the dimer to dissociate; if the medium can trap the resulting excited structure, then the compound exhibits fluorescence in the 410 nm region, which is typical of its protonated or deprotonated forms. PMID- 20017469 TI - Selective recognition of uracil and its derivatives using a DNA repair enzyme structural mimic. AB - During DNA repair, uracil DNA glycosylase (UDG) pulls unwanted uracil into its active site through hydrogen bonding and pi-pi stacking interactions. The reason why UDG binds only uracil tightly--and not its derivatives, such as thymine- remains unclear. In this study, we synthesized the stable, water-soluble receptor 1a as a structural mimic of the active site in UDG. Compound 1a contains a 2,6 bis(glycylamino)pyridine group, which mimics the amino acid residues of UDG that interact with uracil through a hydrogen-bonding network; it also possesses a pyrene moiety as a pi-pi stacking interaction element and fluorescent probe that mimics the aromatic groups (phenyl and fluorescent indolyl units) found in the active site of UDG. Receptor 1a binds selectively to uracil and derivatives (including thymine, 5-formyluracil, 5-fluorouracil, and 5-nitrouracil) and some DNA and RNA nucleosides (including thymidine and uridine) through hydrogen bonding and pi-pi stacking interactions. Interestingly, a plot of log K(b) with respect to the values of pK(a) of the N(3)H units of uracil and its derivatives was linear, with a negative slope (beta) of -0.24 +/- 0.03. Thus, compounds featuring lower values of pK(a) for their N(3)H units provided greater apparent binding constants for their complexes with receptor 1a, suggesting acidity dependent binding of uracil and its derivatives to this receptor; notably, uracil bound more tightly than did thymine. Our study provides some insight into how uracil and its derivatives in DNA are bound by DNA repair enzymes through hydrogen bonding and pi-pi stacking interactions. PMID- 20017470 TI - Heteroaryl cross-coupling as an entry toward the synthesis of lavendamycin analogues: a model study. AB - ABC analogues of the antitumor antibiotic lavendamycin, which contain the key metal chelation site and redox-active quinone unit essential for biological activity, were prepared via the palladium(0)-catalyzed cross-coupling reaction of various 2-haloheteroaromatics with 2-stannylated pyridines and quinolines. Using the Stille reaction, 2-bromo substituted quinolines and 1-bromoisoquinolines were found to undergo efficient coupling with 2-pyridinylstannanes to provide unsymmetrical heterobiaryl derivatives. While the Stille reaction using the reverse coupling partners (i.e., 2-quinolinylstannanes and haloheteroaromatics) had not received much attention in the literature, we found that this alternative coupling reaction efficiently provided several new heterobiaryl derivatives. The gold-catalyzed intramolecular cycloisomerization of N-(prop-2-ynyl)-1H-indole-2 carboxamide smoothly afforded a beta-carbolinone derivative that was subsequently used for a Pd(0)-catalyzed cross-coupling directed toward the synthesis of lavendamycin analogues. PMID- 20017472 TI - An experimental and theoretical study on the remarkable influence of protecting groups on the selectivity of addition of amines to vinyl sulfone-modified hex-2 enopyranosides. AB - Although phenylmethylene-protected vinyl sulfone-modified carbohydrate 2alpha reacts with both primary and secondary amines in Michael fashion to afford aminated products, only primary amines react with the dibenzyl-protected 3alpha, 6-O-trityl-protected 4alpha, and unprotected 5alpha, highlighting for the first time the remarkable influence of protecting groups on the reaction patterns of vinyl sulfone-modified carbohydrates. The quantum chemical calculations suggest that the Michael addition of amines and proton transfer to vinyl sulfone-modified carbohydrates 2alpha and 5alpha are possible via relay process in a concerted mechanism. These calculations reveal that the addition of primary amines to vinyl sulfone-modified carbohydrate is preferential due to the low activation energy barriers, whereas the addition of secondary amines has relatively higher activation energy barriers. The theoretical conclusions are in line with the experimental observations. PMID- 20017471 TI - Dynamics of liquid plugs of buffer and surfactant solutions in a micro-engineered pulmonary airway model. AB - We describe a bioinspired microfluidic system that resembles pulmonary airways and enables on-chip generation of airway occluding liquid plugs from a stratified air-liquid two-phase flow. User-defined changes in the air stream pressure facilitated by mechanical components and tuning the wettability of the microchannels enable generation of well-defined liquid plugs. Significant differences are observed in liquid plug generation and propagation when surfactant is added to the buffer. The plug flow patterns suggest a protective role of surfactant for airway epithelial cells against pathological flow-induced mechanical stresses. We discuss the implications of the findings for clinical settings. This approach and the described platform will enable systematic investigation of the effect of different degrees of fluid mechanical stresses on lung injury at the cellular level and administration of exogenous therapeutic surfactants. PMID- 20017473 TI - Supercritical fluids for the fabrication of semiconductor devices: emerging or missed opportunities? PMID- 20017474 TI - Online fluorescence suppression in modulated Raman spectroscopy. AB - Label-free chemical characterization of single cells is an important aim for biomedical research. Standard Raman spectroscopy provides intrinsic biochemical markers for noninvasive analysis of biological samples but is often hindered by the presence of fluorescence background. In this paper, we present an innovative modulated Raman spectroscopy technique to filter out the Raman spectra from the fluorescence background. The method is based on the principle that the fluorescence background does not change whereas the Raman scattering is shifted by the periodical modulation of the laser wavelength. Exploiting this physical property and importantly the multichannel lock-in detection of the Raman signal, the modulation technique fulfills the requirements of an effective fluorescence subtraction method. Indeed, once the synchronization and calibration procedure is performed, minimal user intervention is required, making the method online and less time-consuming than the other fluorescent suppression methods. We analyze the modulated Raman signal and shifted excitation Raman difference spectroscopy (SERDS) signal of 2 mum-sized polystyrene beads suspended in a solution of fluorescent dye as a function of modulation rate. We show that the signal-to noise ratio of the modulated Raman spectra at the highest modulation rate is 3 times higher than the SERDS one. To finally evaluate the real benefits of the modulated Raman spectroscopy, we apply our technique to Chinese hamster ovary cells (CHO). Specifically, by analyzing separate spectra from the membrane, cytoplasm, and nucleus of CHO cells, we demonstrate the ability of this method to obtain localized sensitive chemical information from cells, away from the interfering fluorescence background. In particular, statistical analysis of the Raman data and classification using PCA (principal component analysis) indicate that our method allows us to distinguish between different cell locations with higher sensitivity and specificity, avoiding potential misinterpretation of the data obtained using standard background procedures. PMID- 20017475 TI - Mono and tetranuclear gold(I) complexes of tris(1-benzylimidazole-2-yl)phosphine. AB - The reaction of tris(1-benzylimidazole-2-yl)phosphine, (Bzim)(3)P, 1, with Ph(3)AsAuCl in 1:1 stoichiometric ratio produced (Bzim)(3)PAuCl, 2. The reaction of (Bzim)(3)PAuCl with NaAuCl(4) in 1:1 stoichiometry in dichloromethane gives an orange-yellow crystalline tetranuclear gold(I) cluster [{mu-N,N' (Bzim)(3)PAuCl}(2)Au(2)][AuCl(2)][AuCl(4)], 3. Complex 4, [{mu-N,N' (Bzim)(3)PAuCl}(2)Au(2)][AuCl(2)](2) is formed when the reaction stoichiometry of (Bzim)(3)PAuCl and AuCl(4)(-) is 2:1. The crystal structure of 3 shows the formation of a 12-membered macrocycle with Au...Au distances of approximately 3.0 A. The structures of (Bzim)(3)PAuCl and 3 show Au...H-C interactions ranging from 2.57 to 2.95 A. Complex 2 crystallizes in the monoclinic space group P2(1)/n (Z = 4), a = 9.1927(5), b = 13.528(2), c = 22.995(2) A, and beta = 94.537(5)(o). Complex 3 crystallizes in the monoclinic space group P2(1)/c (Z = 4), a = 13.785(4), b = 21.426(6), c = 25.203(8) A, and beta = 96.51(6)degrees. PMID- 20017476 TI - Biogenic scorodite crystallization by Acidianus sulfidivorans for arsenic removal. AB - Scorodite is an arsenic mineral with the chemical formula FeAsO(4)*2H(2)O. It is the most common natural arsenate associated with arsenic-bearing ore deposits. In the present study we show that the thermoacidophilic iron-oxidizing archaeon Acidianus sulfidivorans is able to precipitate scorodite in the absence of any primary minerals or seed crystals, when grown on 0.7 g L(-1) ferrous iron (Fe(2+)) at 80 degrees C and pH 1 in the presence of 1.9 g L(-1) arsenate (H(3)AsO(4)). The simultaneous biologically induced crystallization of ferric iron (Fe(3+)) and arsenic to scorodite prevented accumulation of ferric iron. As a result, crystal growth was favored over primary nucleation which resulted in the formation of highly crystalline biogenic scorodite very similar to the mineral scorodite. Because mineral scorodite has a low water solubility and high chemical stability, scorodite crystallization may form the basis for a novel method for immobilization of arsenic from contaminated waters with high arsenic concentrations. PMID- 20017477 TI - Cyclometalation using d-block transition metals: fundamental aspects and recent trends. PMID- 20017478 TI - Non-heme iron oxygenases generate natural structural diversity in carbapenem antibiotics. AB - Carbapenems are a clinically important antibiotic family. More than 50 naturally occurring carbapenam/ems are known and are distinguished primarily by their C-2/C 6 side chains where many are only differentiated by the oxidation states of these substituents. With a limited palette of variations the carbapenem family comprises a natural combinatorial library, and C-2/C-6 oxidation is associated with increased efficacy. We demonstrate that ThnG and ThnQ encoded by the thienamycin gene cluster in Streptomyces cattleya oxidize the C-2 and C-6 moieties of carbapenems, respectively. ThnQ stereospecifically hydroxylates PS-5 (5) giving N-acetyl thienamycin (2). ThnG catalyzes sequential desaturation and sulfoxidation of PS-5 (5), giving PS-7 (7) and its sulfoxide (9). The enzymes are relatively substrate selective but are proposed to give rise to the oxidative diversity of carbapenems produced by S. cattleya, and orthologues likely function similarly in allied streptomyces. Elucidating the roles of ThnG and ThnQ will focus further investigations of carbapenem antibiotic biosynthesis. PMID- 20017479 TI - Metal complexation and biodegradation of EDTA and S,S-EDDS: a density functional theory study. AB - The complexation reactions of ethylenediaminetetraacetate (EDTA(4-)) and S,S ethylenediaminedisuccinate (S,S-EDDS(4-)) with Co(2+), Ni(2+), Cu(2+), Zn(2+), and Cd(2+) cations are investigated by using the DFT/B3LYP method. The hydration reaction of each metal ion with solvent water cluster is considered with a mixed cluster/continuum model. The subsequent metal complexation is treated as a substitution reaction of the coordinated water molecules by the amino polycarboxylic acid ligand. Thermodynamic cycles are schemed to evaluate the free energy changes for both hydration and complexation processes. The values of complexation free energy changes show that the stabilities of metal complexes with the isomeric ligands follow the order of [M(S,S-EDDS)](2-) in trans(O(6)) conformer < [M(S,S-EDDS)](2-) in trans(O(5))-conformer < [M(EDTA)](2-), implying that the failure to observe the trans(O(6)) conformer under experimental conditions is attributed to its inherent instability. The same trend appears in our steric strain analysis on the various chelate rings in complexes. Because the [M(S,S-EDDS)](2-) in trans(O(6)) complex is not available, we focus on the other two series of complexes with concerned metal ions. The stabilities decrease in the order Cu(2+) > Ni(2+) > Co(2+) > Zn(2+) > Cd(2+) and Cu(2+) > Ni(2+) > Co(2+) > Cd(2+) > Zn(2+) for trans(O(5))-[M(S,S-EDDS)](2-) and [M(EDTA)](2-), respectively. These two tendencies are shown to be consistent with the decrease in the metal-to-ligand charge transfer. Meanwhile, a good quantitative correlation is found between the complexation free energies and the dipole moments for all complexes (excluding the case of Cu(2+)). The far-infrared spectra are present to investigate the characteristics of metal-dependent vibrations, and the further natural bond orbital (NBO) method is taken to show the nature of metal-ligand bonding interactions. Finally, and perhaps most importantly, degrading products of both amino polycarboxylic acids are successfully predicted through calculating the dissociation energies of all C-N bonds in free EDTA, S,S-EDDS, and their successive products. Degradation mechanisms with intramolecular hydrogen transfer are proposed for the biodegradation reactions of S,S-EDDS and its product N-(2-aminoethyl) aspartic acid (AEAA). PMID- 20017480 TI - The role of the substrate lipid in processive glycan polymerization by the peptidoglycan glycosyltransferases. AB - The peptidoglycan glycosyltransferases (PGTs) catalyze the processive polymerization of a C55 lipid-linked disaccharide (Lipid II) to form peptidoglycan, the main component of the bacterial cell wall. Our ability to understand this reaction has been limited due to challenges identifying the appropriate substrate analogues to selectively interrogate the donor (the elongating strand) and acceptor (Lipid II) sites. To address this problem, we have developed an assay using synthetic substrates that can discriminate between the donor and acceptor sites of the PGTs. We have shown that each site has a distinct lipid length preference. We have also established that processive polymerization depends on the length of the lipid attached to the donor. PMID- 20017481 TI - Characterization of anthocyanins and anthocyanidins in purple-fleshed sweetpotatoes by HPLC-DAD/ESI-MS/MS. AB - Purple-fleshed sweetpotatoes (PFSP) can be a healthy food choice for consumers and a potential source for natural food colorants. This study aimed to identify anthocyanins and anthocyanidins in PFSP, and to evaluate the effect of thermal processing on these polyphenolic compounds. Freeze-dried powder of raw and steamed samples of three PFSP varieties were extracted with acidified methanol using a Dionex ASE 200 accelerated solvent extractor. Seventeen anthocyanins were identified by HPLC-DAD/ESI-MS/MS for Stokes Purple and NC 415 varieties with five major compounds: cyanidin 3-caffeoylsophoroside-5-glucoside, peonidin 3 caffeoylsophoroside-5-glucoside, cyanidin 3-caffeoyl-p-hydroxybenzoylsophoroside 5-glucoside, peonidin 3-caffeoyl-p-hydroxybenzoyl-sophoroside-5-glucoside, and peonidin-caffeoyl-feruloylsophoroside-5-glucoside. Okinawa variety showed 12 pigments with 3 major peaks identified as cyanidin 3-caffeoylsophoroside-5 glucoside, cyanidin 3-(6'',6'''-dicaffeoylsophoroside)-5-glucoside and cyanidin 3 (6''-caffeoyl-6'''-feruloylsophoroside)-5-glucoside. Steam cooking had no significant effect on total anthocyanin content or the anthocyanin pigments. Cyanidin and peonidin, which were the major anthocyanidins in the acid hydrolyzed extracts, were well separated and quantified by HPLC with external standards. Cyanidin and peonidin, which contribute to the blue and red hues of PFSP, can be simply quantified by HPLC after acid hydrolysis of the anthocyanins. PMID- 20017482 TI - Self-assembled tetranuclear palladium catalysts that produce high molecular weight linear polyethylene. AB - The phosphine-bis-arenesulfonate ligand PPh(2-SO(3)Li-4-Me-Ph)(2) (Li(2)[OPO]) coordinates as a kappa(2)-P,O chelator in Li[(Li-OPO)PdMe(Cl)] (2a) and (Li OPO)PdMe(L) (L = pyridine (2b); MeOH (2d); 4-(5-nonyl)pyridine) (py', 3)). 2a reacts with AgPF(6) to form {(Li-OPO)PdMe}(n) (2c). Photolysis of 2d yields {(OPO)Pd}(2) (5) in which the [OPO](2-) ligand coordinates as a kappa(3)-O,P,O pincer. 3 self-assembles into a tetramer in which four (Li-OPO)PdMe(py') units are linked by Li-O bonds that form a central Li(4)S(4)O(12) cage. The Pd centers are equivalent but are spatially separated into two identical pairs. The Pd-Pd distance within each pair is 6.04 A. IR data (upsilon(ArSO(3)(-)) region) suggest that the solid state structures of 2a-c are similar to that of 3. 3 reacts with the cryptand Krypt211 to form [Li(Krypt211)][(OPO)PdMe(py')] (4). 3 is in equilibrium with a monomeric (Li-OPO)PdMe(py') species (3') in solution. 2a-c and 3 produce polyethylene (PE) with high molecular weight and a broad molecular weight distribution, characteristic of multisite catalysis. Under conditions where the tetrameric structure remains substantially intact, the PE contains a substantial high molecular weight fraction, while, under conditions where fragmentation is more extensive, the PE contains a large low molecular weight fraction. These results suggest that the tetrameric assembly gives rise to the high molecular weight polymer. In contrast, the monomeric complex 4, which contains a free pendant sulfonate group that can bind to Pd, oligomerizes ethylene to a Schultz-Flory distribution of C(4)-C(18) oligomers. PMID- 20017483 TI - Cyclization of arylacetoacetates to indene and dihydronaphthalene derivatives in strong acids. Evidence for involvement of further protonation of O,O-diprotonated beta-ketoester, leading to enhancement of cyclization. AB - The chemical features, such as substrate stability, product distribution, and substrate generality, and the reaction mechanism of Bronsted superacid-catalyzed cyclization reactions of aromatic ring-containing acetoacetates (beta-ketoesters) were examined in detail. While two types of carbonyl cyclization are possible, i.e., keto cyclization and ester cyclization, the former was found to take place exclusively. The reaction constitutes an efficient method to synthesize indene and 3,4-dihydronapthalene derivatives. Acid-base titration monitored with (13)C NMR spectroscopy showed that the acetoacetates are fully O(1),O(3)-diprotonated at H(0) = -11. While the five-membered ring cyclization of the arylacetoacetates proceeded slowly at H(0) = -11, a linear increase in the rate of the cyclization was found with increasing acidity in the high acidity region of H(0) = -11.8 to 13.3. Therefore, the O(1),O(3)-diprotonated acetoacetates exhibited some cyclizing reactivity, but they are not the reactive intermediates responsible for the acceleration of the cyclization in the high acidity region. The reactive cationic species might be formed by further protonation (or protosolvation) of the O(1),O(3)-diprotonated acetoacetates; i.e., they may be tricationic species. Thermochemical data on the acid-catalyzed cyclization of the arylacetoacetates showed that the activation energy is decreased significantly as compared with that of the related acid-catalyzed cyclization reaction of a compound bearing a single functional group, such as a ketone. These findings indicate that intervention of the trication contributes to the activation of the cyclization of arylacetoacetates in strong acid, and the electron-withdrawing nature of the O protonated ester functionality significantly increases the electrophilicity of the ketone moiety. PMID- 20017484 TI - The hypolipidemic effect of Hibiscus sabdariffa polyphenols via inhibiting lipogenesis and promoting hepatic lipid clearance. AB - Hibiscus sabdariffa extract (HSE) was shown to lower the plasma lipid level and reduce the liver damage. In the present study, we investigated if Hibiscus sabdariffa polyphenols (HPE) exerted a hypolipidemic effect and its putative mechanism on liver. HPE exhibited more potency to decrease plasma cholesterol and LDL cholesterol than the crude extract HSE, and increased HDL cholesterol dose dependently. It decreased the lipid content of hepatocyte through the activation of AMPK and reduction of SREBP-1, thus inhibiting the expression of fatty acid synthase and HMG-CoA reductase. LDLR and LDL binding of HepG2 cells were enhanced when treated with HPE. In conclusion, HPE is worthy of being further investigated and could be developed as an adjunctive for hepatic lipid control and hypolipidemic therapy. PMID- 20017485 TI - Regioselective lipase-catalyzed synthesis of 3-o-acyl derivatives of resveratrol and study of their antioxidant properties. AB - One of the approaches to increasing the bioavailability of resveratrol is to protect its 3-OH phenolic group. In this work, regioselective acylation of resveratrol at 3-OH was achieved by transesterification with vinyl acetate catalyzed by immobilized lipase from Alcaligenes sp. (lipase QLG). The maximum yield of 3-O-acetylresveratrol was approximately 75%, as the lipase also catalyzes its further acetylation affording the diester 3,4'-di-O acetylresveratrol and finally the peracetylated derivative. Long saturated and unsaturated fatty acid vinyl esters were also effective as acyl donors with similar regioselectivity. In contrast, lipase B from Candida antarctica catalyzes the acylation of the phenolic group 4'-OH with 80% yield and negligible formation of higher esters. The analysis of the antioxidant properties showed that the Trolox equivalent antioxidant capability (TEAC) values for the acetyl and stearoyl derivatives at 3-OH were, respectively, 40% and 25% referred to resveratrol. The addition of an acyl chain in the 3-OH position caused a higher loss of activity compared with that at the 4'-OH. PMID- 20017486 TI - Phenolic and short-chained aliphatic organic acid constituents of wild oat (Avena fatua L.) seeds. AB - The objective of this research was to identify and quantify the phenolic and short-chained aliphatic organic acids present in the seeds of three wild-type populations of wild oat and compare these results to the chemical composition of seeds from two commonly utilized wild oat isolines (M73 and SH430). Phenolic acids have been shown to serve as germination inhibitors, as well as protection for seeds from biotic and abiotic stress factors in other species, whereas aliphatic organic acids have been linked to germination traits and protection against pathogens. Wild oat populations were grown under a "common garden" environment to remove maternal variation, and the resulting seeds were extracted to remove the readily soluble and chemically bound phenolic and aliphatic organic acid components. Compounds were identified and quantified using gas chromatography-mass spectrometry. Ferulic and p-coumaric acid comprised 99% of the total phenolic acids present in the seeds, of which 91% were contained in the hulls and 98% were in the chemically bound forms. Smaller quantities of OH benzoic and vanillic acid were also detected. Soluble organic acids concentrations were higher in the M73 isoline compared to SH430, suggesting that these chemical constituents could be related to seed dormancy. Malic, succinic, fumaric and azelaic acid were the dominant aliphatic organic acids detected in all seed and chemical fractions. PMID- 20017487 TI - Inhibitors of oxidation and matrix metalloproteinases, floridoside, and D isofloridoside from marine red alga Laurencia undulata. AB - In the exploration of abundant marine biological resources, edible red alga Laurencia undulata led to two bioactive isolates: floridoside (1) and D isofloridoside (2). For the first time, the antioxidant properties of both derivatives (1 and 2) were characterized via free radical scavenging using the ESR technique, reactive oxygen species (ROS) inhibition, membrane protein oxidation, myeloperoxidase (MPO) inhibition, gene expression levels of glutathione (GSH) and superoxide dismutase (SOD), and protein expression of MMP-2 and MMP-9. The results demonstrate that floridoside and D-isofloridoside possess significant antioxidant capacity and are potential inhibitors of MMP-2 and MMP-9. These results clarified that these components may be responsible for the relative activities of crude extract from this genus, which is used as folk medicine. Furthermore, the structure-activity relationships were also suggested. Both isomers could be effective candidates for applications in food and pharmaceutical fields as natural marine antioxidants. PMID- 20017490 TI - Solubilized derivatives of perylenetetracarboxylic dianhydride (PTCDA) adsorbed on highly oriented pyrolytic graphite. AB - The effect on 2D molecular crystallization caused by the addition of propylthioether side groups to the 3,4,9,10-perylenetetracarboxylic dianhydride (PTCDA) molecule is investigated using scanning tunneling microscopy (STM). The molecule was deposited from 1-phenyloctane onto highly oriented pyrolytic graphite (HOPG) and imaged at the liquid-solid interface. We observe a different structure to previously reported arrangements of PTCDA due to the presence of the propylthioether side groups which inhibits the formation of the herringbone phase. A model, supported by calculations based on density functional theory, is proposed in which molecules form rows stabilized by hydrogen bonding. PMID- 20017489 TI - Potato virus X as a novel platform for potential biomedical applications. AB - We demonstrate that nanoparticles formed from the rod-shaped plant virus Potato virus X (PVX) can serve as a novel platform for biomedical applications. Bioconjugation protocols including amine modification and "click" chemistry allowed the efficient functionalization of PVX with biotins, dyes, and PEGs. Fluorescent-labeled and PEGylated PVX particles revealed that different fluorescent labels have a profound effect on PVX-cell interactions. Applying bioconjugation chemistries to PVX opens the door for chemical functionalization with targeting and therapeutic molecules. PMID- 20017491 TI - Structure-activity relationship and mechanism of action studies of manzamine analogues for the control of neuroinflammation and cerebral infections. AB - Structure-activity relationship studies were carried out by chemical modification of manzamine A (1), 8-hydroxymanzamine A (2), manzamine F (14), and ircinal isolated from the sponge Acanthostrongylophora. The derived analogues were evaluated for antimalarial, antimicrobial, and antineuroinflammatory activities. Several modified products exhibited potent and improved in vitro antineuroinflammatory, antimicrobial, and antimalarial activity. 1 showed improved activity against malaria compared to chloroquine in both multi- and single-dose in vivo experiments. The significant antimalarial potential was revealed by a 100% cure rate of malaria in mice with one administration of 100 mg/kg of 1. The potent antineuroinflammatory activity of the manzamines will provide great benefit for the prevention and treatment of cerebral infections (e.g., Cryptococcus and Plasmodium). In addition, 1 was shown to permeate across the blood-brain barrier (BBB) in an in vitro model using a MDR-MDCK monolayer. Docking studies support that 2 binds to the ATP-noncompetitive pocket of glycogen synthesis kinase-3beta (GSK-3beta), which is a putative target of manzamines. On the basis of the results presented here, it will be possible to initiate rational drug design efforts around this natural product scaffold for the treatment of several different diseases. PMID- 20017492 TI - Structural conservation in band 4.1, ezrin, radixin, moesin (FERM) domains as a guide to identify inhibitors of the proline-rich tyrosine kinase 2. AB - The nonreceptor focal adhesion kinases FAK and Pyk2 play a central role in the regulation of glioma cell proliferation and migration, making them attractive targets to improve clinical outcome. Noncatalytic targeting represents a novel approach to regulate the activity of these tyrosine kinases. A combination of site directed mutagenesis and molecular modeling was used to identify compounds that target the F3 module of the Pyk2 FERM domain. A protein pharmacophore model for the Pyk2 FERM/F3 module, generated utilizing the structural conservation of ligand-bound FERM domains with known 3D structures, was used to search the LeadQuest compound library. Compounds compliant with the model were tested for their ability to inhibit the binding of a monoclonal antibody that maps to a functional site on the F3 module. The highest scoring compound bound directly to the Pyk2 FERM domain, inhibited Pyk2 stimulated glioma migration, and provides the framework for the development of novel therapeutic agents to target the activity of the focal adhesion kinases. PMID- 20017494 TI - Direct stabilization of a phospholipid monolayer on H-terminated silicon. AB - This Article describes a strategy to stabilize a phospholipid monolayer directly on the surface of a H-terminated silicon substrate in order to provide a useful platform for silicon based biosensors. The stabilization of an acrylated phospholipid monolayer is obtained by two-dimensional chain polymerization. As the formation of the lipid monolayer in aqueous solution competes with the oxidation of the silicon surface, several cycles of oxide removal and lipid exposure are necessary to densify the lipid layer. Lipid monolayer formation is followed by Fourier transform infrared spectroscopy. The resulting monolayer is denser than corresponding alkyl monolayers formed on H-terminated silicon via photochemical or thermally initiated reactions. PMID- 20017493 TI - Synthesis and biological evaluation of N-hydroxyphenylacrylamides and N hydroxypyridin-2-ylacrylamides as novel histone deacetylase inhibitors. AB - The histone deacetylases (HDACs) are able to regulate gene expression, and histone deacetylase inhibitors (HDACi) emerged as a new class of agents in the treatment of cancer as well as other human disorders such as neurodegenerative diseases. In the present investigation, we report on the synthesis and biological evaluation of compounds derived from the expansion of a HDAC inhibitor scaffold having N-hydroxy-3-phenyl-2-propenamide and N-hydroxy-3-(pyridin-2-yl)-2 propenamide as core structures and containing a phenyloxopropenyl moiety, either unsubstituted or substituted by a 4-methylpiperazin-1-yl or 4-methylpiperazin-1 ylmethyl group. The compounds were evaluated for their ability to inhibit nuclear HDACs, as well as for their in vitro antiproliferative activity. Moreover, their metabolic stability in microsomes and aqueous solubility were studied and selected compounds were further characterized by in vivo pharmacokinetic experiments. These compounds showed a remarkable stability in vivo, compared to hydroxamic acid HDAC inhibitors that have already entered clinical trials. The representative compound 30b showed in vivo antitumor activity in a human colon carcinoma xenograft model. PMID- 20017495 TI - Catalyst-free synthesis and structural and mechanical characterization of single crystalline Ca2B2O5.H2O nanobelts and stacking faulted Ca2B2O5 nanogrooves. AB - Single crystalline Ca(2)B(2)O(5).H(2)O (CBOH) nanobelts were synthesized for the first time using a facile catalyst-free hydrothermal method at low temperature. After being annealed at 400 degrees C for 10 h, the CBOH nanobelts transformed into curve-shaped Ca(2)B(2)O(5)(CBO) nanogrooves with ordered stacking fault arrays along the [110] direction. The single crystalline CBOH nanobelts and the stacking faulted CBO nanogrooves provided a unique platform for investigating the effects of hydrated H(2)O and stacking faults on the mechanical properties of nanomaterials. Nanoindentation tests were performed directly on individual CBOH nanobelts and CBO nanogrooves to probe their mechanical properties. It was found that the CBOH nanobelts, with the presence of hydrated H(2)O, achieved 28.7% increase in elastic modulus, whereas the stacking faulted CBO nanogrooves, with the absence of hydrated H(2)O, exhibited 99% loss in elastic modulus, compared to bulk CBO. PMID- 20017496 TI - Evaluation of substituted N,N'-diarylsulfonamides as activators of the tumor cell specific M2 isoform of pyruvate kinase. AB - The metabolism of cancer cells is altered to support rapid proliferation. Pharmacological activators of a tumor cell specific pyruvate kinase isozyme (PKM2) may be an approach for altering the classic Warburg effect characteristic of aberrant metabolism in cancer cells yielding a novel antiproliferation strategy. In this manuscript, we detail the discovery of a series of substituted N,N'-diarylsulfonamides as activators of PKM2. The synthesis of numerous analogues and the evaluation of structure-activity relationships are presented as well as assessments of mechanism and selectivity. Several agents are found that have good potencies and appropriate solubility for use as chemical probes of PKM2 including 55 (AC(50) = 43 nM, maximum response = 84%; solubility = 7.3 microg/mL), 56 (AC(50) = 99 nM, maximum response = 84%; solubility = 5.7 microg/mL), and 58 (AC(50) = 38 nM, maximum response = 82%; solubility = 51.2 microg/mL). The small molecules described here represent first-in-class activators of PKM2. PMID- 20017497 TI - Reducing stress on cells with apoferritin-encapsulated platinum nanoparticles. AB - The great potential for medical applications of inorganic nanoparticles in living organisms is severely restricted by the concern that nanoparticles can harmfully interact with biological systems, such as lipid membranes or cell proteins. To enable an uptake of such nanoparticles by cells without harming their membranes, platinum nanoparticles were synthesized within cavities of hollow protein nanospheres (apoferritin). In vitro, the protein-platinum nanoparticles show good catalytic efficiency and long-term stability. Subsequently the particles were tested after ferritin-receptor-mediated incorporation in human intestinal Caco-2 cells. Upon externally induced stress, for example, with hydrogen peroxide, the oxygen species in the cells decreased and the viability of the cells increased. PMID- 20017499 TI - Unprotected vinyl aziridines: facile synthesis and cascade transformations. AB - Functionalized vinylaziridines, readily available from water-stable aziridine aldehydes have led to the construction of a variety of stereochemically rich heterobicycles. A cascade ring-opening/ring-contraction mechanism operates in the course of the process. These results underscore the notion that interesting and useful nitrogen-mediated relay processes can arise when elements of strain are merged with the manifolds of enamine/iminium ion reactivity. PMID- 20017498 TI - "Smart" diblock copolymers as templates for magnetic-core gold-shell nanoparticle synthesis. AB - We report a new strategy for synthesizing temperature-responsive gamma-Fe(2)O(3) core/Au-shell nanoparticles (Au-mNPs) from diblock copolymer micelles. The amphiphilic diblock copolymer chains were synthesized using reversible addition fragmentation chain-transfer (RAFT) with a thermally responsive "smart" poly(N isopropylacrylamide) (pNIPAAm) block and an amine-containing poly(N,N dimethylaminoethylacrylamide) (DMAEAm) block that acted as a reducing agent during gold shell formation. The Au-mNPs reversibly aggregated upon heating the solution above the transition temperature of pNIPAAm, resulting in a red-shifted localized surface plasmon resonance. PMID- 20017500 TI - Polarity-reversible conjugate addition tuned by remote electronic effects. AB - A new concept, polarity-reversible conjugate addition, has been described, based on the findings that the polarity of a classical Michael acceptor can be reversed through remote electronic effects. In addition, the remote electronic effects are tunable, and both five- and six-membered nitrogen rings can be constructed starting from acyclic precursors having the same enone structure unit simply by varying a remote substituent in the molecules. PMID- 20017501 TI - Catalytic enantioselective hydrogenation of N-alkoxycarbonyl hydrazones: a practical synthesis of chiral hydrazines. AB - An enantioselective hydrogenation of hydrazones catalyzed by Rh complexes (Rh Josiphos or Rh-Taniaphos) has been developed. The protocol can be applied to hydrazones with three different protective groups (Boc, Cbz, and methoxycarbonyl), allowing for selective deprotection and further elaboration of the hydrazine products in the presence of other functional groups. PMID- 20017502 TI - Insights from a nanoparticle minuet: two-dimensional membrane profiling through silver plasmon ruler tracking. AB - Individual pairs of polymer-tethered silver nanoparticles, so-called silver plasmon rulers, enable distance and orientation measurements on the nanoscale. The reduced linear dichroism and the spectrum of the light scattered from individual plasmon rulers encode information about their orientation and average interparticle separation, respectively. We took advantage of the gain in information silver plasmon rulers offer as probes in optical tracking and analyzed the translational and rotational motions as well as the extension of individual silver plasmon rulers diffusing on the plasma membrane of lysed HeLa cells. Consistent with a compartmentalization of the cell surface on the length scales of the plasmon rulers, most rulers were either immobilized or performed a confined lateral diffusion. Structural details of a plasmon ruler's confinement region became accessible utilizing the orientation and interparticle separation dependent optical response of the plasmon rulers. This approach, which we refer to as polarization-resolved plasmon coupling microscopy, enabled a detailed structural characterization of individual membrane compartments and provided a quantitative metrics to characterize the structural lateral heterogeneity of cell membranes on submicrometer length scales. In combination with adequate tracking methods, the "dance" performed by membrane confined dimers of flexibly linked noble metal nanoparticles revealed deep insight into the underlying membrane morphology. PMID- 20017503 TI - Rhodium-catalyzed functionalization of sterically hindered alkenes. AB - For the first time the rhodium-catalyzed 1,4-addition of organoboranes to hindered Baylis-Hillman adducts, trisubstituted alkenes, affording highly functionalized alkenes, via addition of the organoboranes and hydroxyelimination, is reported. Moreover, preliminary results have shown that, thanks to the use of a monosubstituted chiral diene ligand, enantio-enriched products were easily accessible, while chiral phosphane ligands were completely inappropriate in this reaction. PMID- 20017504 TI - Structure-function analysis of RAMP1-RAMP3 chimeras. AB - The role of receptor activity modifying protein 1 (RAMP1) in forming receptors with the calcitonin receptor-like receptor (CLR) and the calcitonin receptor (CTR) was examined by producing chimeras between RAMP1 and RAMP3. RAMPs have three extracellular helices. Exchange of helix 1 of the RAMPs or residues 62-69 in helix 2 greatly reduced CLR trafficking (a marker for CLR association). Modeling suggests that these exchanges alter the CLR recognition site on RAMP1, which is more exposed than on RAMP3. Exchange of residues 86-89 of RAMP1 had no effect on the trafficking of CLR but reduced the potency of human (h) alphaCGRP and adrenomedullin. However, these alterations to RAMP1 had no effect on the potency of hbetaCGRP. These residues of RAMP1 lie at the junction of helix 3 and its connecting loop with helix 2. Modeling suggests that the loop is more exposed in RAMP1 than RAMP3; it may play an important role in peptide binding, either directly or indirectly. Exchange of residues 90-94 of RAMP1 caused a modest reduction in CLR expression and a 15-fold decrease in CGRP potency. It is unlikely that the decrease in expression is enough to explain the reduction in potency, and so these may have dual roles in recognizing CLR and CGRP. For CTR, only 6 out of 26 chimeras covering the extracellular part of RAMP1 did not reduce agonist potency. Thus the association of CTR with RAMP1 seems more sensitive to changes in RAMP1 structure induced by the chimeras than is CLR. PMID- 20017505 TI - Quantitative analysis of volatile methylsiloxanes in waste-to-energy landfill biogases using direct APCI-MS/MS. AB - Landfill-biogas utilization is a win-win solution as it creates sources of renewable energy and revenue while diminishing greenhouse gas emissions. However, the combustion of a siloxane-containing biogas produces abrasive microcrystalline silica that causes severe and expensive damages to power generation equipment. Hence, the importance of siloxane analysis of the biogas has increased with the growth of the waste-to-energy market. We have investigated an improved method for the analysis of octamethylcyclotetrasiloxane (D4) and decamethylcyclopentasiloxane (D5) in biogas using deuterated hexamethyldisiloxane (HMDS-d(18)) as an internal standard with direct atmospheric pressure chemical ionization/tandem mass spectrometry (APCI-MS/MS). The use of HMDS-d(18) as a single internal standard provided effective signal compensation for both D4 and D5 in biogas and improved the sensitivity and reliability for the direct APCI MS/MS quantification of these compounds in biogas. Low detection limits ( approximately 2 microg/m(3)) were achieved. The method was successfully applied for the determination of D4 and D5 contents in various samples of biogas recovered for electrical power generation from a landfill site in Montreal. Concentrations measured for D4 and D5 were in the ranges of 131-1275 and 250-6226 microg/m(3), respectively. Among the various landfill zones sampled, a clear trend of decreasing D4 and D5 concentrations was observed for older landfill materials. PMID- 20017506 TI - Cation-controlled crystal growth of silver stearate: cryo-TEM investigation of lithium vs sodium stearate. AB - Cryo-TEM, SAXS, and light microscopy techniques were used to probe the morphology and kinetics of silver stearate self-assembly and crystallization from the reaction of silver nitrate with lithium stearate. Unlike the reaction of sodium stearate with silver nitrate, which proceeds via micelle aggregation, the lithium stearate forms vesicles that drastically change the reaction kinetics of the silver stearate nucleation and self-assembly process. In addition, even with excess silver nitrate present, only about 80% of the lithium stearate can be converted to silver stearate. The presence of the residual lithium stearate inhibits the silver stearate crystal growth process. Consequently, no silver stearate micelle aggregates of any significant size form, unlike the system utilizing sodium stearate. Instead, significantly smaller silver stearate crystals result from lithium stearate compared to the silver stearate crystals from sodium stearate and provide an opportunity to further control silver stearate self-assembly and crystal growth. PMID- 20017507 TI - Control of alpha-alumina surface charge with carboxylic acids. AB - In this work, we studied the surface charge of alpha-alumina treated with carboxylic acids with different carbon chain length. The results show the possibility of controlling surface charges of alumina by using different concentrations of carboxylic acids or changing the size of the carbon chain of the acids. We also report that part of the acid found on the surface is strongly bound, therefore making it possible to obtain pH-resistant samples of alpha alumina with an isoelectric point (IEP) of 5.5. It is found, that IEP values obtained for modified samples have a linear correlation with the number of carbon atoms of dicarboxylic acids for up to five carbon atoms. From a practical perspective, the method presented in this work has many advantages. First, it maintains the same hydrophilicity of the alumina surface. Second, the modification of the surface is stable in a long-range of pH. Finally, the presented method is easy-to-use and cheap, as the modification consists of only two simple steps carried out at low temperatures with inexpensive and nontoxic reagents. PMID- 20017508 TI - Kinetics of catalytic reactions on nanoclusters. AB - Kinetics of heterogeneous catalytic reactions on nanoclusters is considered. The size of organic molecules used in various catalytic transformations is often ca. 0.5-1 nm, whereas nanoparticles in the range 2-4 nm are used in supported heterogeneous catalysts. Proposed quantitative treatment of a heterogeneous catalytic mechanism with two kinetically significant steps assumes that a finite number of molecules per a single metal cluster can be adsorbed. A general equation with polynomial terms in the numerator and the denominator is derived for nonideal (nonuniform) surfaces when all kinetic and adsorption constants depend on the spatial arrangements of reacting molecules. A simplified case of a two-step catalytic cycle is discussed, with the first step being adsorption on nanoclusters and the second one being further transformation of absorbed species. The corresponding kinetic equations are derived and numerically analyzed for the case of ideal (uniform) surfaces as well as the presence of induced nonuniformity, when contrary to the ideal case there are lateral interactions between adsorbed species. The form of kinetic equations allows not only saturation, typical for Eley-Rideal kinetics, but also positive cooperativity, i.e., S-shaped behavior of the reaction rate as a function of substrate concentration. Comparison with experimental data on hydrodechlorination of dichlorobenzenes is given. PMID- 20017509 TI - Assembly of zwitterionic phospholipid/conjugated polyelectrolyte complexes: structure and photophysical properties. AB - We report on the formation of complexes between zwitterionic phospholipid vesicles and an anionic fluorescent conjugated polyelectrolyte and the effect of mono- and divalent cations on the photophysical properties of these complexes. Our goal is to gain an understanding of the interplay of morphology and exciton transport in these complexes, information that is critical to designing efficient lipid/conjugated polymer-based sensors. Our studies further underscore the potential application of lipid/conjugated polymer complexes in light-harvesting devices. Our work focuses on the negatively charged conjugated polyelectrolyte poly[5-methoxy-2-(3-sulfopropoxy)-1,4-phenylenevinylene] (MPS-PPV) and its interaction with the zwitterionic lipid dioleoylphosphatidylcholine (DOPC). We utilize monovalent and divalent cations as a tool to control and explore the interaction of MPS-PPV with lipids. We show that Ca(2+) ions promote the complexation of zwitterionic lipids and MPS-PPV in comparison to Na(+) ions. The addition of increasing amounts of zwitterionic phospholipids in the form of vesicles gradually disrupts MPS-PPV aggregates albeit vesicle structure is preserved in Na(+) buffer. Lipid complexation and the resulting MPS-PPV aggregate disruption produces an intensity enhancement and blue shifting of the MPS-PPV emission peak. In the absence of Ca(2+), the intensity enhancement and blue shift reach a plateau at larger than a 10:1 lipid/MPS-PPV monomer mole ratio. In the presence of Ca(2+), a plateau is reached at equimolar concentrations of MPS-PPV and lipid. Vesicle particle coalescence and agglomerate formation are observed herein. Lipid complexation and concomitant MPS-PPV shielding is shown to diminish the quenching of MPS-PPV emission by water-soluble quencher methyl viologen. FRET experiments conducted with membrane-intercalating acceptor dye DiD further underscore the large lipid/polymer interaction mediated by Ca(2+). We observe efficient light harvesting and MPS-PPV-amplified emission quenching in Ca(2+) buffer and to a lesser extent in Na(+) buffer. Our results highlight how the interplay of a zwitterionic lipid, cations, and buffer, in combination with the conjugated polyelectrolyte MPS-PPV, provides rich diversity in architecture and photophysical properties. PMID- 20017510 TI - Carbenoid insertion into the peroxide bond vs the olefin bond of cyclic peroxides. AB - Herein we report examples of the insertion of a carbenoid into a peroxide linkage. This study reveals that intramolecular insertion of carbenes into the peroxide linkage of 3,6-dihydro-1,2-dioxines is preferred over olefin insertion. The initial scope of the reaction and mechanistic considerations, have been probed. This methodology also generates unusual bicyclic hemiacetals (2) and tricyclic peroxides (3). PMID- 20017511 TI - A facile and controllable strategy to synthesize Au-Ag alloy nanoparticles within polyelectrolyte multilayer nanoreactors upon thermal reduction. AB - A new synthesis strategy has been developed for the preparation of bimetallic gold-silver (Au-Ag) alloy nanoparticles by the virtue of polyelectrolyte multilayer (PEM) nanoreactors. By controlling the assembly conditions, gold and silver ions can be effectively loaded onto the PEM composed of polyethylenimine (PEI) and poly(acrylic acid) (PAA) simultaneously. Upon further thermal treatment, Au-Ag alloy nanoparticles with sizes of ca. 3.8 nm formed in the PEM, which were characterized in detail by UV-vis absorption spectroscopy, transmission electron microscopy (TEM), X-ray photoelectron spectroscopy (XPS), X ray diffraction (XRD), and energy-dispersive X-ray (EDX) analysis. Appearance of a single plasmon band in the visible region and lack of apparent core-shell structures in the TEM images confirm the formation of homogeneous Au-Ag alloy nanoparticles. In addition, the surface plasmon absorption band of the Au-Ag alloy nanoparticles shows linear blue-shift with increasing Ag content, which also supported the formation of alloy nanoparticles. Several key parameters of the present strategy have been investigated, which showed that pH of both the assembly solution and gold salt solution and the choice of polymers for constructing PEM, as well as the reduction approach, all played an important role in successfully synthesizing bimetallic Au-Ag nanoparticles. The formation mechanism of alloy nanoparticles has also been discussed based on the spectral evolution during the thermal reduction. PMID- 20017512 TI - Selective nanofiber deposition using a microfluidic confinement approach. AB - The ability to create structurally complex, accurate micropatterns in biocompatible and biodegradable nanofiber mats is important in maximally mimicking the natural extracellular matrix (ECM) for cell biology and tissue engineering. We present a simple, versatile method for the selective deposition of nanofibers with high definition using a unique microfluidic fiber collector. The collector contains fiber etching solutions that are selectively confined to defined regions by means of photocleavable self-assembled monolayer technology and microfluidic capillary filling. The presented approach achieves arbitrarily shaped, microsized, structurally accurate architectures in both random and aligned nanofibers. PMID- 20017513 TI - Stabilization of polyion complex nanoparticles composed of poly(amino acid) using hydrophobic interactions. AB - We report the design and preparation of polyion complex (PIC) nanoparticles composed of anionic hydrophobically modified and cationic poly(amino acid) and the effect of hydrophobic interactions on the stability of these PIC nanoparticles under physiological conditions. We selected poly(gamma-glutamic acid) (gamma-PGA) as the biodegradable anionic polymer and poly(epsilon-lysine) (epsilon-PL) as the cationic polymer. Amphiphilic graft copolymers consisting of gamma-PGA and L-phenylalanine (L-Phe) as the hydrophobic side chain were synthesized by grafting L-Phe to gamma-PGA. The PIC nanoparticles were prepared by mixing gamma-PGA-graft-L-Phe (gamma-PGA-Phe) with epsilon-PL in phosphate buffered saline (PBS). The formation and stability of the PIC nanoparticles were investigated by dynamic light scattering (DLS) measurements. Monomodal anionic PIC nanoparticles were obtained using nonstoichiometric mixing ratios. When unmodified gamma-PGA was mixed with epsilon-PL in PBS, the formation of PIC nanoparticles was observed. However, within a few hours after the preparation, the PIC nanoparticles dissolved in the PBS. In contrast, gamma-PGA-Phe/epsilon-PL nanoparticles showed high stability for a prolonged period of time in PBS and over a wide range of pH values. The stability and size of the PIC nanoparticles depended on the gamma-PGA-Phe/epsilon-PL mixing ratio and the hydrophobicity of the gamma-PGA. The improved stability of the PIC nanoparticles was attributed to the formation of hydrophobic domains in the core of the nanoparticles. The fabrication of PIC nanoparticles using hydrophobic interactions was very useful for the stabilization of the nanoparticles. These results will provide a novel concept in the design of carrier systems composed of PIC. It is expected that the gamma-PGA-Phe/epsilon-PL nanoparticles will have great potential as multifunctional carriers for pharmaceutical and biomedical applications, such as drug and vaccine delivery systems. PMID- 20017515 TI - Direct observation of roaming radicals in the thermal decomposition of acetaldehyde. AB - The thermal dissociation of acetaldehyde has been studied with the reflected shock tube technique using H(D)-atom atomic resonance absorption spectrometry detection. The use of an unreversed light source yields extraordinarily sensitive H atom detection. As a result, we are able to measure both the total decomposition rate and the branching to radical versus molecular channels. This branching provides a direct measure of the contribution from the roaming radical mechanism since the contributions from the usual tight transition states are predicted by theory to be negligible. The experimental observations also provide a measure of the rate coefficient for H + CH(3)CHO. Another set of experiments employing C(2)H(5)I as an H-atom source provides additional data for this rate coefficient that extends to lower temperature. An evaluation of the available experimental results for H + CH(3)CHO can be expressed by a three-parameter Arrhenius expression as k = 7.66 x 10(-20)T(2.75) exp((-486 K)/T) cm(3) molecule( 1) s(-1) (298-1415 K). Analogous experiments employing C(2)D(5)I as a D-atom source allow for the study of the isotopically substituted reaction. The present experiments are the only direct measure for this reaction rate constant, and the results can be expressed by an Arrhenius expression as k = 5.20 x 10(-10) exp(( 4430 K)/T) cm(3) molecule(-1) s(-1) (1151-1354 K). The H/D + CH(3)CHO reactions are also studied with ab initio transition-state theory, and the results are in remarkably good agreement with the current experimental data. PMID- 20017514 TI - Characterization of DNA damage induced by a natural product antitumor antibiotic leinamycin in human cancer cells. AB - Leinamycin is a structurally novel Streptomyces-derived natural product that displays very potent activity against various human cancer cell lines (IC(50) values in the low nanomolar range). Previous in vitro biochemical studies have revealed that leinamycin alkylates DNA, generates apurinic (AP) sites and reactive oxygen species (ROS), and causes DNA strand breaks. However, it is not clear whether these events occur inside cells. In the present study, we have determined the endogenous amount of AP sites and DNA strand breaks in genomic DNA and the amount of oxidative stress in a human pancreatic carcinoma cell line, MiaPaCa, treated with leinamycin by utilizing the aldehyde-reactive probe assay, the comet assay, and fluorescent probes, respectively. We demonstrated that AP sites are formed rapidly following exposure to leinamycin, and the number of AP sites was increased up to seven-fold in a dose-dependent manner. However, only 25 50% of these sites remain 2 h after media containing drug molecules were aspirated and replaced with fresh media. We also observed leinamycin-induced ROS generation and a concomitant increase in apoptosis of MiaPaCa cells. Because both AP sites and ROS have the potential to generate strand breaks in cellular DNA, the comet assay was utilized to detect damage to nuclear DNA in leinamycin treated MiaPaCa cell cultures. Both alkaline and neutral electrophoretic analysis revealed that leinamycin produces both single- and double-stranded DNA damage in drug-treated cells in a dose-dependent manner. Taken together, the results suggest that rapid conversion of leinamycin-guanine (N7) adducts into AP sites to produce DNA strand breaks, in synergy with leinamycin-derived ROS, accounts for the exceedingly potent biological activity of this natural product. PMID- 20017517 TI - DFT study of intermolecular [4 + 2] versus [3 + 2] cycloadditions in the dimerization of 2,4,6-trinitrotoluene (TNT): regioselectivity and stereoselectivity. AB - Two novel types of intermolecular hetero cycloadditions in the participation of the nitro group are put forward in the dimerization of TNT, in comparison with Diels-Alder cross-linking of benzene ring skeletons. Possible transition states and products; for example, their geometrical details, vibrational frequencies, and energies are verified at the B3LYP/6-31+G(d,p) level. Contrary to the hetero Diels-Alder reaction, the folding of the benzene ring side endo is slightly selective specific in 1,3-dipolar cycloaddition. The substituent pattern on reactivity indicates that the methyl group at the bridged sites significantly retards the reaction. Two new sigma bonds are formed kinetically and thermodynamically through the single state; however, the first sigma bond is more easily generated by the triplet state. The shock-wave-produced chemically bound dimer of TNT is most likely to be the oxygen-carbon linkage product. The initial chemical events in TNT under high pressure can be extended to interpret the shock insensitivity of other unsaturated nitro-explosives. The calculated results agree well with some experimental observations. PMID- 20017516 TI - Development of near-infrared fluorophore (NIRF)-labeled activity-based probes for in vivo imaging of legumain. AB - Asparaginyl endopeptidase, or legumain, is a lysosomal cysteine protease that was originally identified in plants and later found to be involved in antigen presentation in higher eukaryotes. Legumain is also up-regulated in a number of human cancers, and recent studies suggest that it may play important functional roles in the process of tumorigenesis. However, detailed functional studies in relevant animal models of human disease have been hindered by the lack of suitably selective small molecule inhibitors and imaging reagents. Here we present the design, optimization, and in vivo application of fluorescently labeled activity-based probes (ABPs) for legumain. We demonstrate that optimized aza-peptidyl Asn epoxides are highly selective and potent inhibitors that can be readily converted into near-infrared fluorophore-labeled ABPs for whole body, noninvasive imaging applications. We show that these probes specifically label legumain in various normal tissues as well as in solid tumors when applied in vivo. Interestingly, addition of cell-penetrating peptides to the probes enhanced cellular uptake but resulted in increased cross-reactivity toward other lysosomal proteases as the result of their accumulation in lysosomes. Overall, we find that aza-peptidyl Asn ABPs are valuable new tools for the future study of legumain function in more complex models of human disease. PMID- 20017518 TI - Enantioselective palladium(II)-catalyzed formal [3,3]-sigmatropic rearrangement of 2-allyloxypyridines and related heterocycles. AB - Enantioselective palladium(II)-catalyzed formal [3,3]-sigmatropic rearrangement of (E)- and (Z)-allyloxy substituted N-heterocycles generates N-allyl N heterocyclic amides in good yields and high enantioselectivities (up to 96% ee). The chiral palladacycle COP-Cl (5 mol %) is used as a catalyst with silver(I) trifluoroacetate (10 mol %) at 35-45 degrees C. Examples of heterocycles synthesized include 2-pyridones, quinolin-2(1H)-ones, and isoquinolin-1(2H)-ones. PMID- 20017519 TI - Synthetic, cyclic voltammetric, structural, EPR, and UV-Vis spectroscopic studies of thienyl-containing meso-A(2)B-cor(Cr(V)=O) systems: consideration of three interrelated molecular detection modalities. AB - We report eight new A(2)B-type (M(n+)) corrolate compounds (two structural studies) that include the oxo[5,15-bis(pentafluorophenyl)-10-R corrolatochromium(V)] [R = 2-/3-thienyl (1a/2a), 3-thianaphthyl (3a)] species. The first examples of meso-A(2) (thienyl)- and Cr-A(2)B-corrole types are represented herein. Characterization includes cyclic voltammetry, electron paramagnetic resonance, 2D ((1)H and (13)C) NMR, and UV-vis spectroscopy, mass spectrometry, and elemental analysis. Compounds 1a-3a have enabled analyte binding capacity studies. [Cu(2+)...O=Cr(cor)] binding represents a new selective mode of corrole-based detection, whereas free-base A(2)B-corroles exhibited limited M(n+) selectivity. The 10-position substitution affects optical profiles in analyte titrations. A limited amount of PPh(3) O-atom uptake from [O=Cr(cor)] was also demonstrated. PMID- 20017520 TI - Nanoparticle coatings for enhanced capture of flowing cells in microtubes. AB - Recently, a flow-based selectin-dependent method for the capture and enrichment of specific types of cells (CD34+ hematopoetic stem and progenitor cells and human leukemia HL60) from peripheral blood was demonstrated. However, these devices depend on a monolayer of selectin protein, which has been shown to have a maximum binding efficiency as a function of surface area. A novel surface coating of colloidal silica nanoparticles was designed that alters the surface roughness resulting in increased surface area. The nanoparticles were adhered using either an inorganic titanate resinous coating or an organic polymer of poly-L-lysine. Using Alexa Fluor 647 conjugated P-selectin, an increase in protein adsorption of up to 35% when compared to control was observed. During perfusion experiments using P-selectin-coated microtubes, we observed increased cell capture and greatly decreased rolling velocity at equivalent protein concentration compared to nonparticle control. Atomic force microscopy showed increased surface roughness consistent with the nanoparticle mean diameter, suggesting a monolayer of particles. These results support the coating's potential to improve existing cell capture implantable devices for a variety of therapeutic and scientific uses. PMID- 20017521 TI - Regioselective intramolecular dipolar cycloaddition of azides and unsymmetrical alkynes. AB - Enantioenriched allenylsilanes are used in three-component propargylation reactions with aldehydes and silyl ethers to form syn-homopropargylic ethers that contain an imbedded azide. These materials then undergo thermally induced intramolecular 1,3-dipolar cycloaddition reactions, resulting in unique fused ring systems containing 1,2,3-triazoles. The ability to modify all three components of the reaction allows for expedient access to compounds containing significant structural and stereochemical variation. PMID- 20017522 TI - Hexaphenylbenzenes as potential acetylene sponges. AB - Acetylene sponges can be created by taking advantage of the nonplanar geometry of hexaphenylbenzenes and the special capacity of the central aromatic ring to engage in C(sp)-H...pi interactions reinforced by secondary C(sp(2))-H...pi interactions, as revealed by X-ray crystallographic studies and DFT calculations. PMID- 20017523 TI - Interactions of tyrosine in Leu-enkephalin at a membrane-water interface: an ultrafast two-dimensional infrared study combined with density functional calculations and molecular dynamics simulations. AB - The interactions of neuropeptides and membranes play an important role in peptide hormone function. Our current understanding of peptide-membrane interactions remains limited due to the paucity of experimental techniques capable of probing such interactions. In this work, we study the nature of opioid peptide-membrane interactions using ultrafast two-dimensional infrared (2D IR) spectroscopy. The high temporal resolution of 2D IR is particularly suited for studying highly flexible opioid peptides. We investigate the location of the tyrosine (Tyr) side chain of leucine-enkephalin (Lenk) in lipid bilayer membranes by measuring spectral diffusion of the phenolic ring vibrational mode in three different systems: Lenk in lipid bilayer membranes (bicelles), Lenk in deuterated water, and p-cresol in deuterated water. Frequency-frequency correlation functions obtained from waiting-time-dependent 2D IR spectra reveal an ultrafast decaying component with an approximately 1 ps time constant that is common for all three systems. On the basis of density functional theory calculations and molecular dynamics simulations, this spectral diffusion component is attributed to hydrogen bond dynamics of the phenolic hydroxyl group interacting with bulk water. Unlike p-cresol in water, both Lenk systems exhibit static spectral inhomogeneity, which can be attributed to conformational distributions of Lenk that do not interconvert within 4 ps. Our results suggest that the Tyr side chain of Lenk in bicelles is located at the water-abundant region at the membrane-water interface and not embedded into the hydrophobic core. PMID- 20017524 TI - Molecular dynamics simulation of poly(ethylene terephthalate) oligomers. AB - Molecular dynamics simulations of poly(ethylene terephthalate) (PET) oligomers are performed in the isobaric-isothermal (NpT) ensemble at a state point typical of a finishing reactor. The oligomer size ranges from 1 to 10 repeat units. We report thermodynamic properties (density, potential energy, enthalpy, heat capacity, isothermal compressibility, and thermal expansivity), transport properties (self-diffusivity, zero-shear-rate viscosity, thermal conductivity), and structural properties (pair correlation functions, hydrogen bonding network, chain radius of gyration, chain end-to-end distance) as a function of oligomer size. We compare the results with existing molecular-level theories and experimental data. Scaling exponents as a function of degree of polymerization are extracted. The distribution of the end-to-end distance is bimodal for the dimer and gradually shifts to a single peak as the degree of polymerization increases. The scaling exponents for the average chain radius of gyration and end to-end distance are 0.594 and 0.571, respectively. The values of the heat capacity, isothermal compressibility, and thermal expansivity agree well with the available experimental data, which are of much longer PET chains. The scaling exponents for the self-diffusivity and zero-shear-rate viscosity are, respectively, -2.01 and 0.96, with the latter one being close to the theoretical prediction 1.0 for short-chain polymers. PMID- 20017525 TI - pH induced DNA folding at interface. AB - DNA chains with cytidine-enriched sequences have been grafted on a gold surface at different pH values monitored by a quartz crystal microbalance with dissipation (QCM-D) in real time. DNA chains grafted under acidic conditions with i-motif structures have a lower density due to the larger chain coverage. In contrast, DNA chains grafted under basic conditions with extended conformations are more densely grafted. After the grafted DNA chains completely unfold at pH 8.5, we have investigated the DNA folding with lower and higher grafting densities by use of QCM-D and surface plasmon resonance (SPR). We find that DNA chains with low grafting density can fold into i-motif structures, whereas those with higher grafting density only partially fold because crowding provides limited space for their conformational changes. PMID- 20017526 TI - Pure isolation and stabilization of energetically highly disfavored geometric isomers by controlling the stereoselectivity of supramolecular interactions in tailored host-guest systems. AB - Energetically highly disfavored geometric isomers are present in only trace amounts in solution and hence cannot be clearly observed by conventional spectroscopic methods. Here, we provide spectroscopic evidence that a suitably sized/shaped cavitand (alpha-cyclodextrin) can discriminate distinctive stereochemical differences between all possible cis-trans stereoisomers of a qualified compound, showing selective recognition solely for the target unstable isomer. We tested a set of guests, and we were able to obtain, for each one separately, purely thermodynamic selectivity of the host for the energetically highly dispreferred stereoisomer among all other equilibrating geometric isomers under ambient conditions. PMID- 20017527 TI - Long-range ordering in the lyotropic lamellar phase studied by high-resolution magnetic resonance diffusion-weighted imaging. AB - Diffusion-weighted magnetic resonance imaging (DW MRI) was applied to the lyotropic lamellar phase of the dodecylammonium chloride/water system (DDACl/H(2)O). In the course of employing a well-known medical imaging method, namely, diffusion tensor imaging (DTI), the system morphology was assessed accurately in the most straightforward way by two-dimensional visualization of eigenvectors associated with planar distribution of effective diffusion tensors throughout the whole slice with 40 microm in-plane resolution. Long-range order was observed in the studied lamellar phase, and morphology was best described by a combination of three- and one-dimensional diffusion. PMID- 20017528 TI - Magnetic field alignable domains in phospholipid vesicle membranes containing lanthanides. AB - Magnetic fields were applied as a structuring force on phospholipid-based vesicular systems, using paramagnetic lanthanide ions as magnetic handles anchored to the vesicle membrane. Different vesicle formulations were investigated using small angle neutron scattering (SANS) in a magnetic field of up to 8 T, cryo-transmission electron microscopy (cryo-TEM), (31)P NMR spectroscopy, dynamic light scattering (DLS), and permeability measurements with a fluorescent water-soluble marker (calcein). The investigated vesicle formulations consisted usually of 80 mol % of the phospholipid 1-palmitoyl-2 oleoyl-sn-glycero-3-phosphocholine (POPC) and 20 mol % of a chelator lipid (DMPE DTPA; 1,2-dimyristoyl-sn-glycero-3-phosphoethanolamine diethylenetriaminepentaacetate) with complexed lanthanide ions (Tm(3+), Dy(3+), or La(3+)), and the total lipid concentration was 15 mM. Vesicles containing the paramagnetic lanthanide Tm(3+) or Dy(3+) exhibited a temperature-dependent response to magnetic fields, which can be explained by considering the formation of lipid domains, which upon reaching a critical size become alignable in a magnetic field. The features of this "magnetic field alignable domain model" are as follows: with decreasing temperature (from 30 to 2.5 degrees C) solid domains, consisting mainly of the higher melting phospholipid (DMPE-DTPA.lanthanide), begin to form and grow in size. The domains assemble the large magnetic moments conferred by the lanthanides and orient in magnetic fields. The direction of alignment depends on the type of lanthanide used. The domains orient with their normal parallel to the magnetic field with thulium (Tm(3+)) and perpendicular with dysprosium (Dy(3+)). No magnetic field alignable domains were observed if DMPE-DTPA is replaced either by POPE-DTPA (1-palmitoyl-2-oleoyl-sn-glycero-3 phosphoethanolamine-diethylenetriamine-pentaacetate) or by DMPC (1,2-dimyristoyl sn-glycero-3-phosphocholine). PMID- 20017529 TI - Application of Raman microscopy to biodegradable double-walled microspheres. AB - Raman mapping measurements were performed on the cross section of the ternary phase biodegradable double-walled microsphere (DWMS) of poly(D,L-lactide-co glycolide) (50:50) (PLGA), poly(L-lactide) (PLLA), and poly(epsilon-caprolactone) (PCL), which was fabricated by a one-step solvent evaporation method. The collected Raman spectra were subjected to a band-target entropy minimization (BTEM) algorithm in order to reconstruct the pure component spectra of the species observed in this sample. Seven pure component spectral estimates were recovered, and their spatial distributions within DWMS were determined. The first three spectral estimates were identified as PLLA, PLGA 50:50, and PCL, which were the main components in DWMS. The last four spectral estimates were identified as semicrystalline polyglycolic acid (PGA), dichloromethane (DCM), copper phthalocyanine blue, and calcite, which were the minor components in DWMS. PGA was the decomposition product of PLGA. DCM was the solvent used in DWMS fabrication. Copper-phthalocyanine blue and calcite were the unexpected contaminants. The current result showed that combined Raman microscopy and BTEM analysis can provide a sensitive characterization tool to DWMS, as it can give more specific information on the chemical species present as well as the spatial distributions. This novel analytical method for microsphere characterization can serve as a complementary tool to other more established analytical techniques, such as scanning electron microscopy and optical microscopy. PMID- 20017530 TI - Novel three-dimensional pillared-layer Ln(III)-Cu(I) coordination polymers featuring spindle-shaped heterometallic building units. AB - The hydrothermal reaction of rare earth oxide, CuO, 2,6-pyridinedicarboxylic acid, and 4,4'-bipyridine in the presence of H(3)PO(3) resulted in the formation of a new series of 3d-4f heterometallic coordination polymers [Ln(pydc)(3)Cu(3)(bipy)(3).m(H(2)O)](n) (Ln = Pr (1), Nd (2), m = 5; Ln = Sm (3), Eu (4), Gd (5), Tb (6), Er (7), Yb (8), m = 4; pydc = 2,6-pyridinedicarboxylate anion; bipy = 4,4'-bipyridine). Complexes 1-8 are isostructural and structurally characterized by elemental analysis, FT-IR spectroscopy, thermogravimetry differential thermal analysis (TG-DTA), single-crystal X-ray diffraction, X-ray powder diffraction (XRPD), and nitrogen adsorption/desorption techniques. The synthesis results show that the addition of H(3)PO(3) in the reaction plays an important role in the formation of the compounds. Single-crystal X-ray diffraction analysis reveals that the heterometallic ions are first interconnected by mixing bridging ligands to produce a spindle-shaped heterometallic ring [Ln(6)(pydc)(6)Cu(12)(bipy)(6)], which is used as the second building unit (SBUs) and finally pillared by bridging bipy molecules to form the rare 3D pillared-layer porous Ln(III)-Cu(I) coordination polymers. Luminescence measurements made under excitation by UV rays reveal that Sm-Cu, Eu-Cu, and Tb-Cu compounds exhibit the characteristic emission bands of Sm(3+), Eu(3+), and Tb(3+) ions in the visible regions, respectively; near-infrared (NIR) emission bands from Nd(III) and Yb(III) ions can also be obtained in Nd-Cu and Yb-Cu compounds, respectively; while Pr-Cu, Gd-Cu, and Er-Cu compounds all display similar emission spectra of Cu(I) coordination compounds in the visible regions. PMID- 20017531 TI - Determination of ionization energies of small silicon clusters with vacuum ultraviolet radiation. AB - In this work we report on single photon vacuum ultraviolet photoionization of small silicon clusters (n = 1-7) produced via laser ablation of Si. The adiabatic ionization energies (AIE) are extracted from experimental photoionization efficiency (PIE) curves with the help of Franck-Condon simulations, used to interpret the shape and onset of the PIE curves. The obtained AIEs are (all energies are in eV) Si (8.13 +/- 0.05), Si(2) (7.92 +/- 0.05), Si(3) (8.12 +/- 0.05), Si(4) (8.2 +/- 0.1), Si(5) (7.96 +/- 0.07), Si(6) (7.8 +/- 0.1), and Si(7) (7.8 +/- 0.1). Most of the experimental AIE values are in good agreement with density functional electronic structure calculations. To explain observed deviations between the experimental and theoretical AIEs for Si(4) and Si(6), a theoretical search of different isomers of these species is performed. Electronic structure calculations aid in the interpretation of the a(2)Pi(u) state of Si(2)(+) dimer in the PIE spectrum. Time-dependent density functional theory calculations are performed to reveal the energies of electronically excited states in the cations for a number of Si clusters. PMID- 20017532 TI - Stabilization of ion concentration polarization using a heterogeneous nanoporous junction. AB - We demonstrate a recycled ion-flux through heterogeneous nanoporous junctions, which induce stable ion concentration polarization with an electric field. The nanoporous junctions are based on integration of ionic hydrogels whose surfaces are negatively or positively charged for cationic or anionic selectivity, respectively. Such heterogeneous junctions can be matched up in a way to achieve continuous ion-flux operation for stable concentration gradient or ionic conductance. Furthermore, the combined junctions can be used to accumulate ions on a specific region of the device. PMID- 20017533 TI - Resolving single-molecule assembled patterns with superresolution blink microscopy. AB - In this paper we experimentally combine a recently developed AFM-based molecule by-molecule assembly (single-molecule cut-and-paste, SMCP) with subdiffraction resolution fluorescence imaging. Using "Blink-Microscopy", which exploits the fluctuating emission of single molecules for the reconstruction of superresolution images, we resolved SMCP assembled structures with features below the diffraction limit. Artificial line patterns then served as calibration structures to characterize parameters, such as the labeling density, that can influence resolution of Blink-Microscopy besides the localization precision of a single molecule. Finally, we experimentally utilized the adjustability of blink parameters to demonstrate the general connection of photophysical parameters with spatial resolution and acquisition time in superresolution microscopy. PMID- 20017534 TI - Synthesis and in vitro cytotoxicity profile of the R-enantiomer of 3,4 dihydroxymethamphetamine (R-(-)-HHMA): comparison with related catecholamines. AB - (+/-)-3,4-Methylenedioxymethamphetamine (MDMA, also known as "ecstasy") is a chiral drug that is essentially metabolized in humans through O-demethylenation into 3,4-dihydroxymethamphetamine (HHMA). There has recently been a resurgence of interest in the possibility that MDMA metabolites, especially 5-(N-acetylcystein S-yl)-N-methyl-alpha-methyldopamine (designated as 5-NAC-HHMA), might play a role in MDMA neurotoxicity. However, the chirality of MDMA was not considered in previously reported in vivo studies because HHMA, the precursor of the 5-NAC-HHMA metabolite, was used as the racemate. Since the stereochemistry of this chiral drug needs to be considered, the first total synthesis of R-(-)-HHMA is reported. Using L-DOPA as the chiral source, the preparation of R-(-)-HHMA is achieved through seven steps, in 30% overall yield and 99.5% enantiomeric excess. The cytotoxicity of R-(-)-HHMA and related catecholamines has been further determined by flow cytometric analysis of propidium iodide uptake in human dopaminergic neuroblastoma SH-SY5Y cells and by an Escherichia coli plate assay, specific for the detection of oxidative toxicity. The good correlation between the toxicities observed in both systems suggests that SH-SY5Y cells are sensitive to oxidative toxicity and that cell death (necrosis) would be mediated by reactive oxygen species mainly generated from redox active quinonoid centers. In contrast, apoptosis was detected for 3,4-dimethoxymethamphetamine (MMMA), the synthetic precursor of HHMA possessing a protected catechol group. MMMA was not toxic in the bacterial assay, indicating that its toxicity is not related to increased oxidative stress. Finally, we can conclude that there is a need to distinguish the toxicity ascribed to MDMA itself, also bearing a protected catechol moiety, from that depending on MDMA biotransformation leading to catechol metabolites such as HHMA and the thioether conjugates. PMID- 20017535 TI - Inhibition of IAPP and IAPP(20-29) fibrillation by polymeric nanoparticles. AB - The fibrillation process of the islet amyloid polypeptide (IAPP) and its fragment (IAPP(20-29)) was studied by means of Thioflavin T (ThT) fluorescence and transmission electron microscopy in the absence and presence of N isopropylacrylamide:N-tert-butylacrylamide (NiPAM:BAM) copolymeric nanoparticles. The process was found to be strongly affected by the presence of the nanoparticles, which retard protein fibrillation as a function of the chemical surface properties of the nanoparticles. The NiPAM:BAM ratio was varied from 50:50 to 100:0. The nanoparticles with higher fraction of NiPAM imposed the strongest retardation of IAPP and IAPP(20-29) fibrillation. These particles have the strongest hydrogen bonding capacity due to the less bulky N-isopropyl group and thus less steric hindrance of the hydrogen-bonding groups of the nanoparticle polymer backbone. Kinetic fibrillation data, as monitored by ThT fluorescence and supported by surface plasmon resonance experiments, suggest that the peptide is strongly absorbed onto the surface of the nanoparticles. This interaction reduces the concentration of peptide free in solution available to proceed to fibrillation which results in an increased lag time of fibrillation, observed as a delayed onset of ThT fluorescence increase, plus a reduction of the amount of fibrils formed as indicated by the equilibrium values at the end of the fibrillation reaction. For the fragment (IAPP(20-29)), the presence of nanoparticles changes the mechanism of association from monomers to fibrils, by interfering with early oligomeric species along the fibrillation pathway. PMID- 20017536 TI - Total synthesis of bengazole A. AB - The divergent and enantioselective total synthesis of the powerful antifungal marine natural product bengazole A has been achieved. PMID- 20017537 TI - An SOA model for toluene oxidation in the presence of inorganic aerosols. AB - A predictive model for secondary organic aerosol (SOA) formation including both partitioning and heterogeneous reactions is explored for the SOA produced from the oxidation of toluene in the presence of inorganic seed aerosols. The predictive SOA model comprises the explicit gas-phase chemistry of toluene, gas particle partitioning, and heterogeneous chemistry. The resulting products from the explicit gas phase chemistry are lumped into several classes of chemical species based on their vapor pressure and reactivity for heterogeneous reactions. Both the gas-particle partitioning coefficient and the heterogeneous reaction rate constant of each lumped gas-phase product are theoretically determined using group contribution and molecular structure-reactivity. In the SOA model, the predictive SOA mass is decoupled into partitioning (OM(P)) and heterogeneous aerosol production (OM(H)). OM(P) is estimated from the SOA partitioning model developed by Schell et al. (J. Geophys. Res. 2001, 106, 28275-28293 ) that has been used in a regional air quality model (CMAQ 4.7). OM(H) is predicted from the heterogeneous SOA model developed by Jang et al. (Environ. Sci. Technol. 2006, 40, 3013-3022 ). The SOA model is evaluated using a number of the experimental SOA data that are generated in a 2 m(3) indoor Teflon film chamber under various experimental conditions (e.g., humidity, inorganic seed compositions, NO(x) concentrations). The SOA model reasonably predicts not only the gas-phase chemistry, such as the ozone formation, the conversion of NO to NO(2), and the toluene decay, but also the SOA production. The model predicted that the OM(H) fraction of the total toluene SOA mass increases as NO(x) concentrations decrease: 0.73-0.83 at low NO(x) levels and 0.17-0.47 at middle and high NO(x) levels for SOA experiments with high initial toluene concentrations. Our study also finds a significant increase in the OM(H) mass fraction in the SOA generated with low initial toluene concentrations, compared to those with high initial toluene concentrations. On average, more than a 1-fold increase in OM(H) fraction is observed when the comparison is made between SOA experiments with 40 ppb toluene to those with 630 ppb toluene. Such an observation implies that heterogeneous reactions of the second-generation products of toluene oxidation can contribute considerably to the total SOA mass under atmospheric relevant conditions. PMID- 20017538 TI - Uniform mesoporous dye-doped silica nanoparticles decorated with multiple magnetite nanocrystals for simultaneous enhanced magnetic resonance imaging, fluorescence imaging, and drug delivery. AB - Highly versatile nanocomposite nanoparticles were synthesized by decorating the surface of mesoporous dye-doped silica nanoparticles with multiple magnetite nanocrystals. The superparamagnetic property of the magnetite nanocrystals enabled the nanoparticles to be used as a contrast agent in magnetic resonance (MR) imaging, and the dye molecule in the silica framework imparted optical imaging modality. Integrating a multitude of magnetite nanocrystals on the silica surface resulted in remarkable enhancement of MR signal due to the synergistic magnetism. An anticancer drug, doxorubicin (DOX), could be loaded in the pores and induced efficient cell death. In vivo passive targeting and accumulation of the nanoparticles at the tumor sites was confirmed by both T2 MR and fluorescence imaging. Furthermore, apoptotic morphology was clearly detected in tumor tissues of mice treated with DOX loaded nanocomposite nanoparticles, demonstrating that DOX was successfully delivered to the tumor sites and its anticancer activity was retained. PMID- 20017539 TI - Helical structure of xylose-DNA. AB - Synthetic biology and systems chemistry demonstrate a growing interest in modified nucleotides to achieve an enzymatically stable artificial nucleic acid. A potential candidate system is xylose-DNA, in which the 2'-deoxy-beta-D-ribo furanose is substituted by 2'-deoxy-beta-D-xylo-furanose. We present here the helical structure and conformational analysis of xylose-DNA on the basis of 35 ns MD simulations of a 29-base-pair DNA duplex. Starting from a right-handed xylose DNA helix, we observe a remarkable conformational transition from right- to left handed helix. The left-handed xylose-DNA is highly dynamic, involving screwing and unscrewing motion of the helix. The sugar pucker induced helical changes influence the backbone to adopt the backbone angles for xylose-DNA while retaining the Watson-Crick base pairing and stacking interactions. The results demonstrate the chiral orthogonality of the ribose and xylose based episomes. As far as stability and compactness of information storage is concerned, the ribose based natural DNA is unsurpassed. PMID- 20017540 TI - Molecular dynamics simulations of the dimerization of transmembrane alpha helices. AB - Membrane proteins account for nearly a quarter of all genes, but their structure and function remain incompletely understood. Most membrane proteins have transmembrane (TM) domains made up of bundles of hydrophobic alpha-helices. The lateral association of TM helices within the lipid bilayer is a key stage in the folding of membrane proteins. It may also play a role in signaling across cell membranes. Dimerization of TM helices is a simple example of such lateral association. Molecular dynamics (MD) simulations have been used for over a decade to study membrane proteins in a lipid bilayer environment. However, direct atomistic (AT) MD simulation of self-assembly of a TM helix bundle remains challenging. AT-MD may be complemented by coarse-grained (CG) simulations, in which small numbers of atoms are grouped together into particles. In this Account, we demonstrate how CG-MD may be used to simulate formation of dimers of TM helices. We also show how a serial combination of CG and AT simulation provides a multiscale approach for generating and refining models of TM helix dimers. The glycophorin A (GpA) TM helix dimer represents a paradigm for helix helix packing, mediated by a GxxxG sequence motif. It is well characterized experimentally and so is a good test case for evaluating computational methods. CG-MD simulations in which two separate TM helices are inserted in a lipid bilayer result in spontaneous formation of a right-handed GpA dimer, in agreement with NMR structures. CG-MD models were evaluated via comparison with data on destabilizing mutants of GpA. Such mutants increased the conformational flexibility and the dissociation constants of helix dimers. GpA dimers have been used to evaluate a multiscale approach: A CG model is converted to an AT model, which is used as the basis of an AT-MD simulation. Comparison of three AT-MD simulations of GpA, one starting from a CG model and two starting from NMR structures, leads to convergence to a common refined structure for the dimer. CG MD self-assembly has also been used to model dimerization of the TM domain of the syndecan-2 receptor protein. This TM helix contains a GxxxG motif, which mediates right-handed helix packing comparable to that of the GxxxG motif in GpA. The multiscale approach has been applied to a more complex system, the heterodimeric alphaIIb/beta3 integrin TM helix dimer. In CG-MD, both right-handed and left handed structures were formed. Subsequent AT-MD simulations showed that the right handed structure was more stable, yielding a dimer in which the GxxxG motif of the alphaIIb TM helix packed against a hydrophobic surface of the beta3 helix in a manner comparable to that observed in two recent NMR studies. This work demonstrates that the multiscale simulation approach can be used to model simple membrane proteins. The method may be applied to more complex proteins, such as the influenza M2 channel protein. Future refinements, such as extending the multiscale approach to a wider range of scales (from CG through QM/MM simulations, for example), will expand the range of applications and the accuracy of the resultant models. PMID- 20017542 TI - Two-dimensional symmetrical mixtures in an external field of square symmetry. AB - Using the Monte Carlo simulation method in the grand canonical ensemble, we study two-dimensional symmetrical binary mixtures subjected to an external potential of square symmetry and finite corrugation. The results reveal a rich variety of mixed liquid-like and solid-like structures. It is demonstrated that even very weakly corrugated external potential leads to the development of axially ordered striped solid-like phases. The results of finite temperature simulation confirm quite well the predictions stemming from the ground state considerations and show the formation of commensurate, high-order commensurate, and incommensurate mixed phases. It is also shown that the corrugation potential affects the demixing transition. PMID- 20017541 TI - Molecular determinants for PP2A substrate specificity: charged residues mediate dephosphorylation of tyrosine hydroxylase by the PP2A/B' regulatory subunit. AB - Together with protein phosphatase 1, protein phosphatase 2A (PP2A) contributes the bulk of Ser/Thr phosphatase activity in most cell types. The predominant form of PP2A is a heterotrimer of catalytic (C), scaffolding (A), and diverse regulatory subunits (B, B', and B''). We have previously shown that N-terminal phosphorylation sites in tyrosine hydroxylase (TH), the rate-limiting enzyme in catecholamine synthesis, are specifically dephosphorylated by PP2A holoenzymes containing the B'beta regulatory subunit. Here, we identify a Glu residue conserved in B' regulatory subunits that is critical for dephosphorylation and inactivation of tyrosine hydroxylase in vitro and in PC12 cells. According to the PP2A heterotrimer crystal structure, Glu153 (B'beta numbering) abuts the catalytic site on the C subunit, and we demonstrate that Glu153 substitution inhibits multisite TH dephosphorylation without compromising PP2A/B'beta holoenzyme assembly or in vitro dephosphorylation of model substrates. Apart from its role in modulating TH activity, Glu153 is also necessary for PP2A/B'beta mediated enhancement of nerve growth factor signaling. Furthermore, global phosphoproteome analysis suggests that Glu153 mediates dephosphorylation of most B'beta substrates in PC12 cells. With regard to selectivity determinants in the substrate, we show that B'beta Glu153 recognizes Arg37 and Arg38 in TH to direct dephosphorylation of both upstream (Ser31) and downstream (Ser40) sites. These results provide evidence of a subunit-spanning substrate docking site on the PP2A/B' holoenzyme, in which negatively charged side chains in the regulatory subunit interact with positive charges proximal to phosphorylated residues to mediate site-specific dephosphorylation. PMID- 20017543 TI - Molecular property investigations of an ortho-hydroxy Schiff base type compound with the first-principle molecular dynamics approach. AB - The structure, proton transfer, and vibrational dynamics under ambient conditions of a selected ortho-hydroxy Schiff base type compound, 2-(N-methyl-alpha iminoethyl)-4-chlorophenol, containing a very short intramolecular hydrogen bond, were investigated computationally in the gas phase and in the crystal by density functional theory (DFT) based first-principle molecular dynamics (FPMD). It is found that the proton is well localized on the nitrogen side of the O...H...N bridge in the crystal phase, in agreement with X-ray diffraction experiments, while a more labile proton is located most of the time on the oxygen side in a vacuum. Environmental effects on this very strong hydrogen bond thus appear crucial and lead to drastic changes of the infrared (IR) spectrum: The computed gas-phase IR spectrum shows a very broad absorption band that covers frequencies from about 1000 to 3000 cm(-1) assigned to the labile proton. In mere contrast, a much more localized absorption band around 2600-2700 cm(-1) is predicted in the crystal phase. Finally, effects of the quantization of the proton motion on the hydrogen bond structure were estimated in two ways. First, we constructed the one dimensional (1D) potential energy surface (PES) for the proton along the O...H...N bridge in a vacuum. The 1D Schrodinger equation was then solved. Next, path integral molecular dynamics (PIMD) was performed in the solid state. Inclusion of quantum effects does not affect the observed change of the most probable tautomer, upon going from the gas phase to the crystal. PMID- 20017544 TI - Orientational mechanisms in liquid crystalline systems. 2. The contribution to solute ordering from the reaction field interaction between the solute electric quadrupole moment and the solvent electric field gradient. AB - In the previous paper of this issue, [Celebre, G.; Ionescu, A. J. Phys. Chem. B doi: 10.1021/jp907310g], following a generalized reaction field approach in the linear response approximation, we were successful in obtaining an analytical compact expression for the mean-field anisotropic orientational potential U(Q EFG) theoretically experienced by a highly idealized nonionic and apolar solute, considered as a point quadrupole immersed in a uniaxial polarizable continuum medium (model of a nematic solvent comprised of dipolar mesogenic molecules). The term U(Q-EFG) describes the electrostatic interaction between the electric quadrupole of the solute and the electric field gradient induced at the solute by the surrounding medium polarized by the distribution of electric charges representing the quadrupolar solute itself. In the present paper, the obtained potential has been considered as an additional orientational interaction contributing to the solute ordering, besides the well-recognized and very effective "short-range" (size-and-shape-dictated) mechanisms. Since in our theory the solvent is characterized by its dielectric tensor, the model has been widely tested by taking as references the experimental order parameters of several uniaxial and biaxial different small rigid probe molecules (H(2), N(2), acetylene, allene, propyne, benzene, hexafluorobenzene, 1,4-difluorobenzene, and norbornadiene) dissolved in the nematic solvents ZLI1132 (Deltaepsilon >> 0) and EBBA (Deltaepsilon < 0); moreover, the order parameters of the same solutes in the so-called nematic "magic mixture" (45 wt % EBBA + 55 wt % ZLI1132), where the short-range orientational effects are commonly believed to be very dominant, have been conventionally assumed as reference of the absence of electrostatic orientational effects. The experimental order parameters of the treated solutes, obtained in the past by liquid crystal NMR and available from literature, have been then compared with those theoretically predicted by our theoretical approach in order to obtain useful hints about two basic points, (a) the real physical nature of the interactions (other than the "size-and-shape") involved in the orientational mechanisms and (b) the conceptual effectiveness of the suggested mean-field approach in describing this kind of phenomena. Successes and failures of the approach in the predictions are discussed at length, along with their possible reasons and implications. PMID- 20017545 TI - Orientational mechanisms in liquid crystalline systems. 1. A reaction field analytical description of the interaction between the electric quadrupole moment of a probe solute and the electric field gradient of a nematic solvent. AB - In the present paper, the fundamental problem of calculating the electric field gradient (EFG) experienced by a highly idealized solute, represented by a general point quadrupole immersed in an anisotropic uniaxial medium, has been tackled. Following a generalized reaction field approach (based upon the original ideas and the "mean-field philosophy" due to Kirkwood and Onsager) in the linear response approximation, a closed analytical expression of the EFG has been derived (to the best of our knowledge, for the first time). The obtained expression is particularly simple and elegant, also thanks to the oversimplifying approximation that the virtual cavity containing the solute is assumed to be perfectly spherical. This compact and manageable formula, obtained by a rigorous mathematical derivation (unlike other mean-field phenomenological models previously suggested in literature) can be useful to investigate and better understand a likely orientational mechanism, partly responsible for the ordering of small solutes dissolved in nematic mesophases, based on the interaction between the electric quadrupole of the solute and the electric field gradient of the anisotropic uniaxial medium (in the next paper of this issue, the formulation obtained in this work is widely tested on a variety of uniaxial and biaxial solutes dissolved in different nematic solvents). PMID- 20017546 TI - Electrochemical stability of nanometer-scale Pt particles in acidic environments. AB - Understanding and controlling the electrochemical stability or corrosion behavior of nanometer-scale solids is vitally important in a variety of applications such as nanoscale electronics, sensing, and catalysis. For many applications, the increased surface to volume ratio achieved by particle size reduction leads to lower materials cost and higher efficiency, but there are questions as to whether the intrinsic stability of materials also decreases with particle size. An important example of this relates to the stability of Pt catalysts in, for example, proton exchange fuel cells. In this Article, we use electrochemical scanning tunneling microscopy to, for the first time, directly examine the stability of individual Pt nanoparticles as a function of applied potential. We combine this experimental study with ab initio computations to determine the stability, passivation, and dissolution behavior of Pt as a function of particle size and potential. Both approaches clearly show that smaller Pt particles dissolve well below the bulk dissolution potential and through a different mechanism. Pt dissolution from a nanoparticle occurs by direct electro-oxidation of Pt to soluble Pt(2+) cations, unlike bulk Pt, which dissolves from the oxide. These results have important implications for understanding the stability of Pt and Pt alloy catalysts in fuel cell architectures, and for the stability of nanoparticles in general. PMID- 20017547 TI - Conductive behavior of high TiO2 nanoparticle content of inorganic/organic nanostructured composites. AB - Amphiphilic polystyrene-b-poly(ethylene oxide) (PS-b-PEO) diblock copolymers with different block ratios were used as templates for the incorporation of a high content of titanium dioxide nanoparticles using the sol-gel method. Confinement of the inorganic part in the PEO block of the block copolymer allows the generation of nanostructured systems with a high nanoparticle content. As successfully demonstrated using tunneling atomic force microscopy, the investigated systems maintained the conductive properties of the TiO(2) nanoparticles. The obtained results confirmed that with increasing TiO(2) nanoparticle content, the local current value increased up to 15 pA, and this conductivity value strongly depended on the amount of the PEO block in the block copolymer template. Moreover, the results indicated that control of the ratio between the sol-gel and the PEO block allows the design of well-dispersed, conductive inorganic/organic hybrids with high inorganic content. These materials can provide attractive strategies in the field of dye-sensitized solar cells. PMID- 20017548 TI - Electronic structures, hole-doping, and superconductivity of the s = 1, 2, 3, and 4 members of the (Cu,Mo)-12s2 homologous series of superconductive copper oxides. AB - We demonstrate that the T(c) value of superconductive copper oxides does not depend on the distance between two adjacent CuO(2) planes as long as the hole doping level and the immediate (crystal) chemical surroundings of the planes are kept the same. Experimental evidence is accomplished for the homologous series of (Cu,Mo)-12s2, the member phases of which differ from each other by the number (s) of cation layers in the fluorite-structured (Ce,Y)-[O(2)-(Ce,Y)](s-1) block between the CuO(2) planes. X-ray absorption near-edge structure spectroscopy is employed as a probe for the hole states of these phases. The s = 1 member appears to be more strongly doped with holes than other phases of the series and accordingly to possess the highest T(c) value of 87 K. For s > or = 2, unexpectedly, both the CuO(2) plane hole concentration and the value of T(c) (approximately 55 K) remain constant, being independent of s. PMID- 20017549 TI - Pd(II)-catalyzed enantioselective C-H olefination of diphenylacetic acids. AB - Pd(II)-catalyzed enantioselective C-H olefination of diphenylacetic acid substrates has been achieved through the use of monoprotected chiral amino acid ligands. The absolute configuration of the resulting olefinated products is consistent with that of a proposed C-H insertion intermediate. PMID- 20017550 TI - Structure-reactivity relationships: reactions of a 5-substituted aziadamantane in a resorcin[4]arene-based cavitand. AB - The complexation properties of two novel C5-substituted adamantanediazirines within the resorcin[4]arene-based cavitand 4 were investigated in DMSO-d(6), revealing that binding is up to 1.4 kcal/mol stronger for halogenated adamantanediazirines when compared with the unsubstituted species. The thermal behavior of 5-bromo-2-aziadamantane (3) was investigated by DSC analysis as the first representative of the adamantanediazirine family in the neat solid state, as well as encapsulated within the aromatic cavity of cavitand 4. In the solid phase, the reactions of photolytically or thermolytically generated 5-bromo-2 adamantanylidene (11) can be controlled by complexation within cavitand 4. PMID- 20017551 TI - Planar metamaterial analogue of electromagnetically induced transparency for plasmonic sensing. AB - We experimentally demonstrate a planar metamaterial analogue of electromagnetically induced transparency at optical frequencies. The structure consists of an optically bright dipole antenna and an optically dark quadrupole antenna, which are cut-out structures in a thin gold film. A pronounced coupling induced reflectance peak is observed within a broad resonance spectrum. A metamaterial sensor based on these coupling effects is experimentally demonstrated and yields a sensitivity of 588 nm/RIU and a figure of merit of 3.8. PMID- 20017552 TI - Cleavage of both C(sp3)-C(sp2) bonds of alkylidenecyclopropanes: formation of ethylene-osmium-vinylidene complexes. AB - The complex [OsTp(kappa(1)-OCMe)(2)(P(i)Pr(3))]BF(4) [Tp = hydridotris(pyrazolyl)borate] promotes the cleavage of both C(sp(3))-C(sp(2)) bonds of benzylidenecyclopropane and 3-phenylpropylidenecyclopropane to yield the complexes [OsTp(=C=CHR)(eta(2)-CH(2)=CH(2))(P(i)Pr(3))]BF(4) (R = Ph, CH(2)CH(2)Ph). The process is proposed to take place via metallacyclopropene intermediates stabilized by an ethylene chelation assistant. The driving force for the fragmentation is the high stability of the resulting ethylene-Os vinylidene species. PMID- 20017553 TI - Synthesis of the lantibiotic lactocin S using peptide cyclizations on solid phase. AB - Lactocin S is a lantibiotic peptide with potent antibacterial activity against a range of gram-positive bacteria. Because of challenges in obtaining sufficient quantities of this compound from natural sources, the stereochemistry of the lanthionine residues in lactocin S had not been confirmed. This report describes the chemical synthesis of lactocin S on chlorotrityl polystyrene resin in 10% overall yield using intramolecular cyclization to form the lanthionine rings and employing fragment coupling for the two N-terminal residues. This represents the first report of solid-supported synthesis of a naturally occurring lantibiotic. Comparison to lactocin S isolated from Lactobacillus sakei L45 using a combination of HPLC, MS/MS sequencing, bacterial testing, and chiral GC-MS analysis confirmed the initially proposed structure and the stereochemistry of the DL-lanthionine residues. PMID- 20017554 TI - Selectively metallized polymeric substrates by microcontact printing an aluminum(III) porphyrin complex. AB - We report a simple, low-cost method for the fabrication of copper wires and contacts on a wide range of flexible, rigid, and inert polymeric substrates. This method relies on procedures to oxidize the polymeric substrates to form surface bound carboxylic acid groups. Patterning of an aluminum porphyrin ink using microcontact printing results in the formation of an aluminum porphyrin monolayer that is covalently anchored to the oxidized polymer surface via an aluminum carboxylate bond. We characterize this monolayer using ultraviolet-visible absorption spectra, reflection-absorption infrared spectroscopy, and contact angle measurements. Patterned aluminum porphyrin monolayers bind a Pd/Sn colloidal catalyst from solution that subsequently initiates the selective deposition of copper in an electroless plating solution. We demonstrate the fabrication of patterned copper films on a variety of both flexible and rigid polymers with minimum feature sizes of 2 microm over 2 cm(2) substrates. Measurements of electrical resistivity of copper wires fabricated on flexible poly(ethylene naphthalate) (PEN) substrates as a function of the bending radius show no negative impact on electrical performance at bending radii as small as 500 microm. Permanently damaging the PEN substrate by creasing (corresponding to a bending radius of 100 microm) results in only a modest increase in resistivity. PMID- 20017555 TI - Stable holographic gratings with small-molecular trisazobenzene derivatives. AB - We present a series of small-molecular trisazobenzene chromophores, including, for instance, 1,3,5-tris{[4-[4-[(4-cyanophenyl)azo]phenoxy]butyryl]amino}benzene that feature a remarkably stable light-induced orientation in initially amorphous thin-film architectures. It is demonstrated that for optimal performance it is critical to design chemical structures that allow formation of both an amorphous and a liquid-crystalline phase. In the present approach, the liquid-crystalline feature was introduced by inserting decoupling spacers between a trisfunctionalized benzene core and the three azobenzene moieties, as well as adding polar end groups to the latter. To compensate for the deleterious reduction of the glass transition temperature associated with the spacers in the compounds, polar units were incorporated between the benzene core and the side groups. Intriguingly, the molecular glasses that feature a latent liquid crystalline phase display a remarkable "postdevelopment", i.e., an increase of the amplitude of refractive index modulation in holographic experiments after writing of optical gratings is arrested, exceeding 20% for the previously mentioned derivative. Thus, these nonpolymeric, azobenzene-containing compounds presented in this work appear to be attractive candidates for fabrication of stable holographic volume gratings. PMID- 20017556 TI - Binding conformation and kinetics of two pheromone-binding proteins from the Gypsy moth Lymantria dispar with biological and nonbiological ligands. AB - Pheromone-binding proteins (PBPs) in insects can bind various substances and selectively deliver the message of a signal molecule to the downstream components of the olfactory system. This can be achieved either through a ligand-specific conformational change of the C-terminal peptide of the PBP or by selectively binding/releasing the ligand. PBP may also act as a scavenger to protect the sensory neurons from saturating at high ligand doses. We have compared two PBPs from the gypsy moth (PBP1 and PBP2) and their truncated forms (TPBPs), which lack the C-terminal peptide, in this study. Stopped-flow kinetics with N-phenyl-1 naphthylamine (NPN) have revealed a diffusion-controlled collisional step, between PBP and NPN, after which the NPN relocates into a hydrophobic environment. This work supports the hypothesis that binding between PBPs and ligands occurs stepwise. With the method of tryptophan fluorescence quenching, we have shown different local conformational changes around Trp 37, induced by different ligands, manifested in changes of both the steric and electronic environment around the residue. Importantly, we have noticed a significant difference in the changes induced by the biological ligand (the pheromone) and nonbiological ligands. Therefore, we hypothesize that PBP may serve a different function in each kinetic step, displaying a unique P.L conformation. PMID- 20017557 TI - Structural and functional characterization of the monomeric U-box domain from E4B. AB - Substantial evidence has accumulated indicating a significant role for oligomerization in the function of E3 ubiquitin ligases. Among the many characterized E3 ligases, the yeast U-box protein Ufd2 and its mammalian homologue E4B appear to be unique in functioning as monomers. An E4B U-box domain construct (E4BU) has been subcloned, overexpressed in Escherichia coli, and purified, which enabled determination of a high-resolution NMR solution structure and detailed biophysical analysis. E4BU is a stable monomeric protein that folds into the same structure observed for other structurally characterized U-box domain homodimers. Multiple sequence alignment combined with comparative structural analysis reveals substitutions in the sequence that inhibit dimerization. The interaction between E4BU and the E2 conjugating enzyme UbcH5c has been mapped using NMR, and these data have been used to generate a structural model for the complex. The E2 binding site is found to be similar to that observed for dimeric U-box and RING domain E3 ligases. Despite the inability to dimerize, E4BU was found to be active in a standard autoubiquitination assay. The structure of E4BU and its ability to function as a monomer are discussed in light of the ubiquitous observation of U-box and RING domain oligomerization. PMID- 20017559 TI - Diphosphine and diarsine complexes of germanium(II) halides-preparation, spectroscopic, and structural studies. AB - The Ge(II) halide complexes [GeX(2)(L-L)] (L-L = o-C(6)H(4)(PPh(2))(2), o C(6)H(4)(PMe(2))(2), Me(2)P(CH(2))(2)PMe(2); X = Cl, Br, I. L-L = Et(2)P(CH(2))(2)PEt(2); X = Cl or Br. L-L = o-C(6)H(4)(AsMe(2))(2); X = Br or I) and [GeCl(L-L)][GeCl(3)] (L-L = o-C(6)H(4)(AsMe(2))(2)) have been prepared and characterized by IR, (1)H and (31)P{(1)H} NMR spectroscopy, and microanalyses. The crystal structures of [GeX(2){o-C(6)H(4)(PPh(2))(2)}] (X = Cl, Br, I) reveal discrete mononuclear units with a very asymmetric bidentate o-C(6)H(4)(PPh(2))(2) ligand and a bent GeX(2) unit. Those of [GeX(2){o-C(6)H(4)(PMe(2))(2)}] show symmetrically coordinated diphosphine with loosely associated dimer arrangements, formed through long Ge...X bridges between adjacent monomer units. [GeX(2){R(2)P(CH(2))(2)PR(2)}] (R = Me; X = Cl, Br, I. R = Et; X = Cl, Br) all show discrete monomer structures with 2-fold crystallographic symmetry and based upon four-coordinate Ge, with the diphosphine chelating and approximately linear GeX(2) units. [GeI(2){o-C(6)H(4)(AsMe(2))(2)}] involves significant intermolecular Ge...I interactions, giving rise to a zigzag polymer chain. Finally, the structure of [GeCl{o-C(6)H(4)(AsMe(2))(2)}][GeCl(3)] shows pyramidal cations and anions both with crystallographic mirror symmetry, with the diarsine symmetrically chelating, and long Ge...Cl interactions give a loosely associated chain polymer with alternating cations and anions. Comparisons across this series of structurally diverse complexes are discussed. PMID- 20017560 TI - Significant effect of bromo substituents on nonlinear optical properties of polymer and chromophores. AB - Four chromophores containing bromo substituents, a ployimide with bromo containing chromophores, four reference functional polyimides, and fourteen reference chromophores were synthesized for studying effect of bromo substituents on nonlinear optical (NLO) properties of materials and chromophores. The results of hyper-Rayleigh scattering and UV-vis spectra show that static first molecular hyperpolarizability (beta(0)) values of bromo-containing chromophores are 1.24 5.75 times as beta(0) of the corresponding chloro-containing chromophores (Hammett constants sigma of chloro and bromo groups are same) without causing a visible shift of the absorption band to longer wavelength. UV-vis spectra and the results of Maker Fringe method show that the polyimide with chromophores containing bromo substituents exhibits a good optical transparency and a much higher macroscopic nonlinear optical coefficient (d(33) = 20.1 pm/V) than the reference polyimides containing nitro (d(33) = 9.6 pm/V) and cyano (d(33) = 8.9 pm/V) groups in spite of nitro and cyano groups being strong electron acceptors. d(33) of polyimide with chloro-containing chromophores is very small. Therefore, this paper suggests an effective strategy for improving the NLO properties of polymeric materials and chromophores without reducing optical transparency in designing NLO polymers and chromophores. On the basis of quantum chemistry calculations, the reasons of effect of bromo substituents on NLO properties of chromophores and materials were discussed. PMID- 20017558 TI - Label-free fluorescent aptamer sensor based on regulation of malachite green fluorescence. AB - We report a label-free fluorescent aptamer sensor for adenosine based on the regulation of malachite green (MG) fluorescence, with comparable sensitivity and selectivity to other labeled adenosine aptamer-based sensors. The sensor consists of free MG, an aptamer strand containing an adenosine aptamer next to an MG aptamer, and a bridging strand that partially hybridizes to the aptamer strand. Such a hybridization prevents MG from binding to MG aptamer, resulting in low fluorescence of MG in the absence of adenosine. Addition of adenosine causes the adenosine aptamer to bind adenosine, weakening the hybridization of the aptamer strand with the bridging strand, making it possible for MG to bind to the aptamer strand and exhibit high fluorescence intensity. Since this design is based purely on nucleic acid hybridization, it can be generally applied to other aptamers for the label-free detection of a broad range of analytes. PMID- 20017561 TI - Time-lapse atomic force microscopy observations of the morphology, growth rate, and spontaneous alignment of nanofibers containing a peptide-amphiphile from the hepatitis G virus (NS3 protein). AB - Time-lapse atomic force microscopy is used in this contribution to directly watch the growth of nanofibers of a lipidated peptide on a mica surface. Specifically, the studied lipopeptide is the palmitoyl derivative of the fragment 505-514 of NS3 protein from the hepatitis G virus, abbreviated as Palmitoyl-NS3 (505-514). Data on the morphology, growth rate, and orientation of these peptide-amphiphile nanofibers have been obtained. From these data, it can be concluded that this synthetic lipopeptide forms two types of fiber-like aggregates: (i) half spherical fibrous aggregates with lengths of hundreds of nanometers and (ii) spherical fibrous aggregates with lengths of several micrometers. In addition, when a fresh lipopeptide aqueous solution is deposited onto a mica surface, the aggregates spontaneously orient parallel to each other, yielding well-aligned nanofibers on large areas of the mica surface. A significant growth in both the length and the number of the fibers was observed during the first minutes after the solution deposition. Elongation of the fibrous aggregates from one end is more frequent, though elongation from both ends also occurs, with growth rates in the 4-5 nm/s range. The effects of dilution, mechanical perturbation, and pH on the aggregation behavior of Palmitoyl-NS3 (505-514) are also detailed in this paper. PMID- 20017562 TI - Defects responsible for the hole gas in Ge/Si core-shell nanowires. AB - The origin of the ballistic hole gas recently observed in Ge/Si core-shell nanowires has not been clearly resolved yet, although it is thought to be the result of the band offset at the radial interface. Here we perform spin-polarized density-functional calculations to investigate the defect levels of surface dangling bonds and Au impurities in the Si shell. Without any doping strategy, we find that Si dangling bond and substitutional Au defects behave as charge traps, generating hole carriers in the Ge core, while their defect levels are very deep in one-component Si nanowires. The defect levels lie to within 10 meV from or below the valence band edge for nanowires with diameters larger than 33 A and the Ge fractions above 30%. As carriers are spatially separated from charge traps, scattering is greatly suppressed, leading to the ballistic conduction, in good agreement with experiments. PMID- 20017563 TI - New type of magnetic tunnel junction based on spin filtering through a reduced symmetry oxide: FeCo|Mg3B2O6|FeCo. AB - Magnetic tunnel junctions with high-tunneling magnetoresistance values such as Fe|MgO|Fe capitalize on spin filtering in the oxide region based on the band symmetry of incident electrons. However, these structures rely on magnetic leads and oxide regions of the same cubic symmetry class. A new magnetic tunnel junction (FeCo|Mg(3)B(2)O(6)|FeCo) is presented that uses a reduced symmetry oxide region (orthorhombic) to provide spin filtering between the two cubic magnetic leads. Complex band structure analysis of Mg(3)B(2)O(6) based on density functional calculations shows that significant spin filtering could occur in this system. This new type of magnetic tunnel junction may have been fabricated already and can explain recent experimental studies of rf-sputtered FeCoB|MgO|FeCoB junctions where there is significant B diffusion into the MgO region. PMID- 20017564 TI - Suppression of auger processes in confined structures. AB - We explore how the size and shape of the microscopic confinement potential affects the nonradiative Auger decay rate of confined carriers. Calculations conducted in the two-band, effective mass Kane model unambiguously show that smoothing out the confinement potential could reduce the rate by more than 3 orders of magnitude relative to the rate in structures with abruptly terminating boundaries. As the confinement potential width is increased, the calculated rate decreases overall, exhibiting very deep minima at regular widths. Such minima suggest that nanocrystals of "magic sizes" can exist for which nonradiative Auger processes are strongly suppressed. PMID- 20017565 TI - Directional photofluidization lithography for nanoarchitectures with controlled shapes and sizes. AB - Highly ordered metallic nanostructures have attracted an increasing interest in nanoscale electronics, photonics, and spectroscopic imaging. However, methods typically used for fabricating metallic nanostructures, such as direct writing and template-based nanolithography, have low throughput and are, moreover, limited to specific fabricated shapes such as holes, lines, and prisms, respectively. Herein, we demonstrate directional photofluidization lithography (DPL) as a new method to address the aforementioned problems of current nanolithography. The key idea of DPL is the use of photoreconfigurable polymer arrays to be molded in metallic nanostructures instead of conventional colloids or cross-linked polymer arrays. The photoreconfiguration of polymers by directional photofluidization allows unprecedented control over the sizes and shapes of metallic nanostructures. Besides the capability for precise control of structural features, DPL ensures scalable, parallel, and cost-effective processing, highly compatible with high-throughput fabrication. Therefore, DPL can expand not only the potential for specific metallic nanostructure applications but also large-scale innovative nanolithography. PMID- 20017566 TI - High-yield synthesis and structure of double-walled bismuth-nanotubes. AB - A new convenient room-temperature template-free route for high-yield synthesis of double-walled bismuth nanotubes through the treatment of solid bismuth monoiodide with n-butyllithium is presented. Scanning electron microscopy and transmission electron microscopy observations of the product show uniform one-dimensional nanoparticles with high aspect ratios and lengths up to several hundred nanometers. Investigations of the cross sections of the bismuth nanotubes reveal an inner diameter of about 4.5 nm and an outer diameter of 6 nm. The tube walls consist of two coaxial cylinders, and the estimated thickness of the double-wall of about 0.75 nm matches quite properly two layers in the rhombohedral bismuth bulk structure. PMID- 20017567 TI - Photoisomerization and thermal isomerization of shuttlecock- and bowl-equipped (phenylazo)pyridines. AB - The photoisomerization and thermal isomerization of sterically hindered (phenylazo)pyridine derivatives with a shuttlecock-shaped (TbetNNPy) or a bowl shaped framework (BmtNNPy as well as the azoxy counterpart, BmtNN(O)Py) have been investigated. The crystal structures of these compounds revealed a planar conformation of the (phenylazo)pyridine moiety for TbetNNPy and severely distorted conformations for BmtNNPy and BmtNN(O)Py. The quantum yields of the trans-to-cis photoisomerization of TbetNNPy and BmtNNPy is lower than those of unsubstituted (phenylazo)pyridines. The low quantum yields may, to a large part, be attributed to electronic factors rather than steric factors. While the shuttlecock framework in TbetNNPy does not affect the thermal cis-to-trans isomerization, as the activation parameters for TbetNNPy are quite similar to those of azobenzene and azopyridine derivatives, the bowl framework in BmtNNPy renders the thermal isomerization process slower by lowering the frequency factor to an extent that more than compensates for the lowered activation energy. The process is characterized with a large negative activation entropy and a small activation enthalpy, implying that the isomerization proceeds through a limited range of intermediates stabilized by the presence of the bowl framework. PMID- 20017568 TI - Luminescence switching by hybridization-directed mixed lanthanide complex formation. AB - We have developed a homogeneous assay method in which the lanthanide ion carrier and light absorbing components of a luminescent lanthanide chelate are separated in two distinct molecules that can together form a luminescent mixed chelate complex. The separated label moieties were conjugated to oligonucleotides which were used as probes to detect a complementary target DNA. The background signal of the assay was very low, indicating the signal was highly dependent on the hybridization of the two probes on adjacent positions on the target oligonucleotide. PMID- 20017569 TI - Thermus thermophilus proteins that are differentially expressed in response to growth temperature and their implication in thermoadaptation. AB - As a kind of important extremophiles to realize the adaptation of life at high temperatures, thermophiles have attracted extensive studies. However, the pathways of thermophile proteins related to thermoadaptation remain to be addressed. Our study showed that there existed two types of protein profiles for the thermophile Thermus thermophilus wl in response to temperature change. One of them came from cultures growing below 65 degrees C, which was close to the optimal growth temperature, and another from cultures at or above 65 degrees C. These protein profiles were confirmed by Northern blots. On the basis of the proteomic and computational analyses, it was found that the thermophile proteins related to thermoadaptation might be involved in metabolic pathways as well as the stabilities and modifications of DNA and proteins. Interestingly, for the basic metabolism glycolysis, the phosphoglucomutase was up-regulated at below optimum temperature, while the glyceraldehyde-3-phosphate dehydrogenase was up regulated at above-optimum temperature, suggesting that different regulations might be used for basic metabolism at different temperatures. To characterize the proteins in response to high temperatures, superoxide dismutase (SOD), an important enzyme in organism to remove free radical produced in stress environment such as high temperature, was selected as a target protein for this investigation. SOD was inactivated to construct a SOD mutant. The results showed that the SOD protein was essential in thermoadaptation of T. thermophilus. Our study, therefore, presented the thermophile proteins required for thermoadaptation and their possible pathways in thermoadaptation. PMID- 20017570 TI - Chemistry or biology? The debate continues... PMID- 20017574 TI - Creating an interdisciplinary introductory chemistry course without time intensive curriculum changes. PMID- 20017575 TI - Wrestling with native chemical ligation. AB - An improved method for the semisynthesis of a potassium channel involving native chemical ligation allows the introduction of short sequences containing non canonical amino acids at any position within the polypeptide chain. The work enhances the technology available for a range of fundamental investigations of membrane proteins and for applications of membrane channels and pores in biotechnology. PMID- 20017576 TI - In silico toxicology challenges for pharmaceuticals: complacency or controversy? PMID- 20017577 TI - The use of computer models in pharmaceutical safety evaluation. AB - With the ever increasing volume of data available to scientists in drug discovery and development, the opportunity to leverage an increasing amount of these data in the assessment of drug safety is clear. The challenge in an environment of increasing data volume is in the structuring and the analysis of these data, such that decisions can be made without excluding information or overstating their meaning. Informatics and modelling play a crucial role in addressing this challenge in two basic ways: a) the data are structured and analysed in a transparent and objective way; and b) new experiments are designed with the model as part of the design process, much like modern experimental physics. Enhancing the use and impact of informatics and modelling on drug discovery is not simply a matter of increasing processor speed and memory capacity. The transformation of raw data to usable, and useful, information is a scientific, technical and, perhaps most importantly, cultural challenge within drug discovery. This review will highlight some of the history, current approaches and promising future directions in this rapidly expanding area. PMID- 20017578 TI - In silico toxicology in drug discovery - concepts based on three-dimensional models. AB - Animal testing is still compulsory worldwide, for the approval of drugs and chemicals produced in large quantities. Computer-assisted (in silico) technologies are considered to be efficient alternatives to in vivo experiments, and are therefore endorsed by many regulatory agencies, e.g. for use in the European REACH initiative. Advantages of in silico methods include: the possible study of hypothetical compounds; their low cost; and the fact that such virtual experiments are typically based on human data, thus making the question of interspecies transferability obsolete. Since the mid-1990s, computer-based technologies have become an indispensable tool in drug discovery - used primarily to identify small molecules displaying a stereospecific and selective binding to a regulatory macromolecule. Since toxic effects are still responsible for some 20% of the late-stage failures, there is a continuing need for in silico concepts which can be used to estimate a compound's ADMET (adsorption, distribution, metabolism, elimination, toxicity) properties - in particular, toxicity. The aim of this paper is to provide an insight into computational technologies that allow for the prediction of toxic effects triggered by pharmaceuticals. As most adverse and toxic effects are mediated by unwanted interactions with macromolecules involved in biological regulatory systems, we have focused on methodologies that are based on three-dimensional models of small molecules binding to such entities, and discuss the results at the molecular level. PMID- 20017579 TI - Physiologically-based simulation modelling for the reduction of animal use in the discovery of novel pharmaceuticals. AB - The global pharmaceutical industry is estimated to use close to 20 million animals annually, in in vivo studies which apply the results of fundamental biomedical research to the discovery and development of novel pharmaceuticals, or to the application of existing pharmaceuticals to novel therapeutic indications. These applications of in vivo experimentation include: a) the use of animals as disease models against which the efficacy of therapeutics can be tested; b) the study of the toxicity of those therapeutics, before they are administered to humans for the first time; and c) the study of their pharmacokinetics - i.e. their distribution throughout, and elimination from, the body. In vivo pharmacokinetic (PK) studies are estimated to use several hundred thousand animals annually. The success of pharmaceutical research currently relies heavily on the ability to extrapolate from data obtained in such in vivo studies to predict therapeutic behaviour in humans. Physiologically-based modelling has the potential to reduce the number of in vivo animal studies that are performed by the pharmaceutical industry. In particular, the technique of physiologically based pharmacokinetic (PBPK) modelling is sufficiently developed to serve as a replacement for many in vivo PK studies in animals during drug discovery. Extension of the technique to incorporate the prediction of in vivo therapeutic effects and/or toxicity is less well-developed, but has potential in the longer term to effect a significant reduction in animal use, and also to lead to improvements in drug discovery via the increased rationalisation of lead optimisation. PMID- 20017580 TI - The in chemico-in silico interface: challenges for integrating experimental and computational chemistry to identify toxicity. AB - A number of toxic effects are brought about by the covalent interaction between the toxicant and biological macromolecules. In chemico assays are available that attempt to identify reactive compounds. These approaches have been developed independently for pharmaceuticals and for other nonpharmaceutical compounds. The assays vary widely in terms of the macromolecule (typically a peptide) and the analytical technique utilised. For both sets of methods, there are great opportunities to capture in chemico information by using in silico methods to provide computational tools for screening purposes. In order to use these in chemico and in silico methods, integrated testing strategies are required for individual toxicity endpoints. The potential for the use of these approaches is described, and a number of recommendations to improve this extremely useful technique, in terms of implementing the Three Rs in toxicity testing, are presented. PMID- 20017581 TI - Computational toxicology approaches at the US Food and Drug Administration. AB - For over a decade, the United States Food and Drug Administration (US FDA) has been engaged in the applied research, development, and evaluation of computational toxicology methods used to support the safety evaluation of a diverse set of regulated products. The basis for evaluating computational toxicology methods is multi-factorial, including the potential for increased efficiency, reduction in the numbers of animals used, lower costs, and the need to explore emerging technologies that support the goals of the US FDA's Critical Path Initiative (e.g. to make decision support information available early in the drug review process). The US FDA's efforts have been facilitated by agency approved data-sharing agreements between government and commercial software developers. This commentary review describes former and current scientific initiatives at the agency, in the area of computational toxicology methods. In particular, toxicology-based QSAR models, ToxML databases and knowledgebases will be addressed. Notably, many of the computational toxicology tools available are commercial products - however, several are emerging as non-commercial products, which are freely-available to the public, and which will facilitate the understanding of how these programs work and avoid the "black box" paradigm. Through productive collaborations, the US FDA Center for Drug Evaluation and Research, and the Center for Food Safety and Applied Nutrition, have worked together to evaluate, develop and apply these methods to chemical toxicity endpoints of regulatory interest. PMID- 20017582 TI - Definition of the applicability domains of knowledge-based predictive toxicology expert systems by using a structural fragment-based approach. AB - The applicability domain of a (quantitative) structure-activity relationship ([Q]SAR) must be defined, if a model is to be used successfully for toxicity prediction, particularly for regulatory purposes. Previous efforts to set guidelines on the definition of applicability domains have often been biased toward quantitative, rather than qualitative, models. As a result, novel techniques are still required to define the applicability domains of structural alert models and knowledge-based systems. By using Derek for Windows as an example, this study defined the domain for the skin sensitisation structural alert rule-base. This was achieved by fragmenting the molecules within a training set of compounds, then searching the fragments for those created from a test compound. This novel method was able to highlight test chemicals which differed from those in the training set. The information was then used to designate chemicals as being either within or outside the domain of applicability for the structural alert on which that training set was based. PMID- 20017583 TI - The impact of introducing computer-based alternatives to the use of animals in the teaching of physiology and pharmacology at Balkan universities - a pilot study. AB - Balkan universities use a substantial number of small mammals and amphibians in the teaching of physiology and pharmacology. This project investigated whether making computer-based alternatives readily available, and combining this availability with a staff development workshop focusing on methods of integrating such resources into undergraduate curricula, would have any effect on animal use. Teachers from 20 Institutes (from five Balkan countries) participated in the workshop. They presented information about animal use in teaching in their universities, and agreed to introduce at least one computer-based alternative into their teaching in the following year. They were surveyed by questionnaire before, during, and one year after, attending the workshop, in order to estimate any changes in animal use. The results showed a significant (P < 0.01) reduction in animal use and a high level of implementation of the alternatives provided at the workshop. Teachers recognised the potential benefits of using computers to support their teaching. They lacked knowledge about what computer-based alternatives are available and how to find information about them, including published evidence of their educational effectiveness. In this pilot study, a combination of staff development and making alternatives readily available to teachers had a significant impact on animal use in the teaching of physiology and pharmacology. PMID- 20017584 TI - The use of mechanisms and modes of toxic action in integrated testing strategies: the report and recommendations of a workshop held as part of the European Union OSIRIS Integrated Project. AB - This report on The Potential of Mode of Action (MoA) Information Derived from Non testing and Screening Methodologies to Support Informed Hazard Assessment, resulted from a workshop organised within OSIRIS (Optimised Strategies for Risk Assessment of Industrial Chemicals through Integration of Non-test and Test Information), a project partly funded by the EU Commission within the Sixth Framework Programme. The workshop was held in Liverpool, UK, on 30 October 2008, with 35 attendees. The goal of the OSIRIS project is to develop integrated testing strategies (ITS) fit for use in the REACH system, that would enable a significant increase in the use of non-testing information for regulatory decision making, and thus minimise the need for animal testing. One way to improve the evaluation of chemicals may be through categorisation by way of mechanisms or modes of toxic action. Defining such groups can enhance read-across possibilities and priority settings for certain toxic modes or chemical structures responsible for these toxic modes. Overall, this may result in a reduction of in vivo testing on organisms, through combining available data on mode of action and a focus on the potentially most-toxic groups. In this report, the possibilities of a mechanistic approach to assist in and guide ITS are explored, and the differences between human health and environmental areas are summarised. PMID- 20017585 TI - Improving the availability of human tissues for medical research in the UK. PMID- 20017586 TI - Myristic acid-conjugated polyethylenimine for brain-targeting delivery: in vivo and ex vivo imaging evaluation. AB - To investigate the potential of myristic acid (MC) to mediate brain delivery of polyethylenimine (PEI) as a gene delivery system, a covalent conjugate (MC-PEI) of MC, and PEI was synthesized. A near-infrared fluorescence probe, IR820 was conjugated to MC-PEI to explore its in vivo distribution after intravenous (i.v.) administration in mice. The brain targeting ability of MC-PEI was evaluated by near-infrared fluorescence imaging and analyzed semiquantitatively by fluorescence intensity, respectively. Significant NIR fluorescent signal was detected in the brain 12 h after i.v. administration and further confirmed by imaging the whole brain and brain slices. Semiquantitative results from fluorescence intensity further supported the successful brain delivery of MC-PEI which led to a very significant increase ( approximately 200%) in the brain uptake after i.v. injection in comparison with unmodified PEI. The capability of MC-PEI to condense DNA was further confirmed using agarose gel retardation assay, indicating its potential for gene delivery. The significant in vivo and ex vivo results suggest that MC-PEI is a promising brain-targeting drug delivery system, especially for gene delivery. PMID- 20017587 TI - An overview of IL-7 biology and its use in immunotherapy. AB - Interleukin (IL)-7 is required for T-cell development as well as for the survival and homeostasis of mature T-cells. In the thymus, the double negative (DN) CD4(-) CD8(-) thymocyte progenitor transition into double positive CD4+ CD8+ cells requires Notch and IL-7 signaling. Importantly, IL-7 seems to have a dose effect on T-cell development and, at high doses, DN progression is blocked. Naive T cells in the thymus, and after their exit to the periphery, are dependent on IL-7 and TCR signaling for survival. Upon antigen exposure, they proliferate and differentiate into memory T-cells. Because IL-7 intervenes at all stages of T cell development and maintenance, it has been introduced recently into clinical trials as an immunotherapeutic agent for cancer patients (of particular note, those who had undergone T-cell depleting therapy) in an attempt to increase their population sizes of CD4+ and CD8+ cells overall, and specifically of CD8+ (CD45RA+)CCR7+ and/or CD27+), CD4+ (CD45RA+CD31+), and CD4+ central memory T cells (CD45RA(-)CCR7+). Interestingly, IL-7 in humans induced a preferential expansion of naive T-cells, resulting in a broader T-cell repertoire than before the treatment; this effect was independent of age. This suggests that IL-7 therapy could enhance immune responses in patients with limited naive T-cell numbers as in aged patients or after disease-induced or iatrogenic T-cell depletion. This overview highlights the role of IL-7 on T-cells in mice and humans. PMID- 20017588 TI - Stuttered and fluent speakers' heart rate and skin conductance in response to fluent and stuttered speech. AB - BACKGROUND: Previous studies have found simultaneous increases in skin conductance response and decreases in heart rate when normally fluent speakers watched and listened to stuttered speech compared with fluent speech, suggesting that stuttering induces arousal and emotional unpleasantness in listeners. However, physiological responses of persons who stutter observing stuttering and fluent speech has not been measured. Research suggests that the mechanism responsible for listeners' reactions is the mirror neuron system. The mirror neuron system activates when perceiving and producing goal-directed actions and forms the link between action perception and action production. AIMS: To compare physiological responses elicited via fluent and stuttered speech in fluent and persons who stutter groups. METHODS & PROCEDURES: Participants included 15 adult persons who stutter (mean age = 29.7 years) and 21 normally fluent controls (mean age = 23.7 years). Participants watched and listened to six speech samples read by three persons who stutter speakers and three fluent speakers, while their skin conductance response and heart rate were recorded simultaneously. Participants' responses to each speech sample were defined as changes in mean heart rate and mean skin conductance response between 15 s before and 15 s during stimuli presentation. OUTCOMES & RESULTS: Both groups showed similar patterns of response: skin conductance response was significantly increased and heart rate was significantly decreased in response to stuttered versus fluent speech. Similar to previous findings, the highest skin conductance response measures were induced by the first presentation of stuttered speech, with the response attenuating on subsequent presentations, while the heart rate was stable over time. CONCLUSIONS & IMPLICATIONS: Results confirmed past research that showed arousal and emotional unpleasantness when observing stuttering. Clinically, these results acknowledge the arousal and emotional discomfort in fluent speakers and persons who stutter. The physiological responses to stuttering and the underlying emotional aspects should be discussed in the therapeutic milieu with notions about coping strategies. PMID- 20017589 TI - Role of transfusion in stem cell transplantation: a freedom-from-transfusion (FFT), cost and survival analysis. AB - BACKGROUND: Transfusion of blood products is often necessary for patients undergoing stem cell transplantation (SCT). The need for red cell and platelet transfusion may vary significantly depending on the type of transplantation and underlying disease. METHODS: In an attempt to evaluate the need and volume of transfusions in patients undergoing SCT at University of Kansas Medical Center, the authors retrospectively evaluated the transfusion data of all patients who received SCT between 2000 and 2005. RESULTS: A total of 138 (90%) out of 154 patients undergoing autologous SCT and 24 (43%) out of 56 patients with allogeneic SCT exhibited total hematopoietic engraftment and freedom from transfusion (FFT). Time to achieve FFT (median; range) for RBC units for autologous SCT (12; 0-183) was significantly shorter compared with allogeneic SCT (16.5; 0-373). Number of RBC units (median; range) transfused were significantly less in patients undergoing autologous SCT (4; 0-26) compared to patients undergoing allogeneic SCT (6.5; 0-54). The median cost of transfusion was significantly higher in patients undergoing allogeneic SCT (red cell: $2,015; platelet: $4,480) compared to patients undergoing autologous SCT (red cell: $1,240; platelet: $2,520). The authors recognize that this was a retrospective single-center study and practice guidelines may vary from center to center. CONCLUSION: Authors conclude that transfusion of blood products is an expensive but integral part of SCT, more so for allogeneic SCT than for patients undergoing autologous SCT. Total FFT is a desirable long-term goal of successful marrow transplantation. PMID- 20017590 TI - Toxic effects of mercuric sulfide on immune organs in mice. AB - Mercuric sulfide (HgS) is a major component of cinnabar, which has been used as a sedative drug in China for more than 2000 years. Because its toxicological effects are still unclear, we attempted to verify the toxic effects of HgS, focused on liver and immune organs such as the spleen and thymus. Male ICR mice were administered HgS (0.02, 0.2, 2.0 g/kg/day) by gavage for 4 weeks. During the administration period, HgS-treated mice did not reveal overt signs of clinical toxicity. HgS had no significant effect on body weight, food consumption, water consumption, and organ weights. In spite of its known insolubility, HgS was absorbed by the gastrointestinal tract and accumulated in the liver, spleen and thymus in a dose-dependent manner. In the biochemical and histological examination, HgS did not cause hepatotoxicity. However, HgS significantly increased both CD8(+) T lymphocytes and CD4(+)CD8(+) lymphocyte populations in the spleen without changing in the thymus. In the histological evaluation, HgS induced enlargement with marked hyperplasia and increase of lymphoid follicles in the spleen. In addition, HgS induced the gene expression of pro-inflammatory cytokines in the spleen and thymus. Our results suggest that insoluble HgS was absorbed by the gastrointestinal tract, accumulated in the spleen and thymus, and thus could affect immune systems. PMID- 20017591 TI - Seasonal variation in chemical composition of size-segregated urban air particles and the inflammatory activity in the mouse lung. AB - We investigated the seasonal variations in the chemical composition and in vivo inflammatory activity of urban air particulate samples in four size ranges (PM(10 2.5), PM(2.5-1), PM(1-0.2), and PM(0.2)). The samples were collected in Helsinki using a high-volume cascade impactor (HVCI). Healthy C57BL/6J mice were intratracheally instilled with a single dose (10 mg/kg) of the particulate samples. The lungs were lavaged and the bronchoalveolar lavage fluid (BALF) was assayed for indicators of inflammation and tissue damage: cytokines (tumor necrosis factor [TNF]-alpha, interleukin [IL]-6, and keratinocyte-derived chemokine [KC]) at 4 h, and total cell number and total protein concentration at 12 h. The PM(10-2.5) and PM(2.5-1) samples had much higher inflammatory potency than the PM(1-0.2) and PM(0.2) samples. The relative inflammatory activities of the autumn samples were the highest on an equal mass basis, but when estimated for the particulate mass per cubic meter of air, the springtime samples had the highest inflammatory potential. Resuspended soil material and other non-exhaust particulate material from traffic were associated with a high inflammatory activity of the PM(10-2.5) and PM(2.5-1) samples. Secondary inorganic ions in the PM(1-0.2) and PM(0.2) samples had inconsistent negative or positive correlations with the inflammatory activity. There were no systematic seasonal variations in the tracers of incomplete combustion and atmospherically oxidized organics in the PM(1-0.2) and PM(0.2) samples, which probably explains their low correlations with the inflammatory activity. In conclusion, in a relatively clean Nordic city, the resuspension of road dust and other non-exhaust particulate material from traffic were the major sources of inflammatory activity of urban air inhalable particles. PMID- 20017592 TI - Increased lung resistance after diesel particulate and ozone co-exposure not associated with enhanced lung inflammation in allergic mice. AB - Exposure to diesel exhaust particulate matter (DEP) exacerbates asthma. Likewise, similar effects have been reported with exposure to the oxidizing air pollutant ozone (O(3)). Since levels of both pollutants in ambient air tend to be simultaneously elevated, we investigated the possible synergistic effect of these agents on the exacerbation of allergic airways disease in mice. Male BALB/c mice were sensitized ip with ovalbumin (Ova) or vehicle only, then exposed once per week for 4 wk via nose-only inhalation (4 h) to the PM(2.5) fraction of DEP (2 mg/m(3)), O(3) (0.5 ppm), DEP and O(3), or filtered air, and then challenged with aerosolized ovalbumin. Ova sensitization in air-exposed mice enhanced pulmonary inflammatory cell infiltration, several indicators of injury in the lung (lactate dehydrogenase, albumin and total protein), and lung resistance (R(L)) and elastance (E(L)) in response to methacholine (MCh) aerosol challenge. DEP exposure did not enhance the Ova-induced increase in pulmonary cell infiltration, indicators of injury, or R(L) and E(L). O(3) exposure enhanced the Ova-induced increase in inflammatory cell infiltration and N-acetylglucosaminidase (NAG) in the lung, but had no effect on R(L) or E(L). DEP co-exposure significantly attenuated the O(3)-induced increase in cell infiltration and indicators of injury; co-exposure had no effect on E(L) relative to air-exposed Ova-sensitized mice. However, only DEP-O(3) co-exposure significantly increased the Ova-induced increase in R(L). Thus, O(3) and DEP co-exposure exacerbated airways hyperresponsiveness, a response that was not associated with parallel increases in pulmonary inflammation and one that may be mediated by a unique mechanism. PMID- 20017593 TI - A transport model for nicotine in the tracheobronchial and pulmonary region of the lung. AB - Nicotine in mainstream cigarette smoke is predominantly present in the particulate phase. Interestingly, however, the deposition efficiency of smoke particles in the respiratory tract is less effective than is the nicotine retention. In the literature, four nicotine deposition mechanisms are identified: (a) direct gas deposition, (b) evaporative gas deposition, (c) particle deposition with evaporation, and (d) particle deposition with diffusion. In this article we present a physically motivated fundamental model to address nicotine deposition mechanisms (b) and (c) from the vapor phase. The model incorporates nicotine mass transport through estimates for the diffusion time across the epithelial layer and the time for nicotine vapor diffusion from the gas volume to the tissue surfaces in the tracheobronchial and pulmonary regions of the respiratory tract. The model comprises four mass transfer processes for nicotine at the surface of the respiratory tract epithelium: (1) conversion of free base nicotine from protonated nicotine; (2) free base nicotine transport across the epithelium; (3) free base nicotine evaporation; and (4) diffusion of free base nicotine vapor from the surface gas layer into the airway lumen. Results of the nicotine mass transport model suggest that the principal mechanism of nicotine delivery to the lung is by direct deposition of particles to the alveolar fluid lining, followed rapidly by evaporation into the lumen and then gas diffusion back to the surface as nicotine depletes in the surface layer through its transport across the epithelium. PMID- 20017594 TI - Pharmacokinetics of radiolabeled tungsten ((188)W) in male Sprague-Dawley rats following acute sodium tungstate inhalation. AB - Aerosol cloud formation may occur when certain tungsten munitions strike hard targets, placing military personnel at increased risk of exposure. Although the pharmacokinetics of various forms of tungsten have been studied in animals following intravenous and oral administration, tungsten disposition following inhalation remains incompletely characterized. The objective of this study was to evaluate the pharmacokinetics of inhaled tungstate (WO(4)) in rats. Male, 16-wk old, CD rats (n = 7 rats/time point) underwent a single, 90-min, nose-only exposure to an aerosol (mass median aerodynamic diameter [MMAD] 1.50 mum ) containing 256 mg W/m(3) as radiolabeled sodium tungstate (Na(2)(188)WO(4)). (188)W tissue concentrations were determined at 0, 1, 3, 7, and 21 days postexposure by gamma spectrometry. The thyroid and urine had the highest (188)W levels postexposure, and urinary excretion was the primary route of (188)W elimination. The pharmacokinetics of tungsten in most tissues was best described with a two-compartment pharmacokinetic model with initial phase half-lives of approximately 4 to 6 h and a longer terminal phase with half-lives of approximately 6 to 67 days. The kidney, adrenal, spleen, femur, lymph nodes, and brain continued to accumulate small amounts of tungsten as reflected by tissue:blood activity ratios that increased throughout the 21-day period. At day 21 all tissues except the thyroid, urine, lung, femur, and spleen had only trace levels of (188)W. Data from this study can be used for development and refinement of pharmacokinetic models for tungsten inhalation exposure in environmental and occupational settings. PMID- 20017595 TI - Murine pulmonary inflammation model: a comparative study of anesthesia and instillation methods. AB - Various techniques have been utilized historically to generate acute pulmonary inflammation in the murine system. Crystalline silica exposure results in acute inflammation followed by pulmonary fibrosis. Methods of exposure are varied in their techniques, as well as types of anesthesia. Therefore, the current study sought to compare the effects of two major anesthesia (isoflurane and ketamine) and three routes of instillation, intranasal (IN), intratracheal (IT), and trans oral (TO), on markers of inflammation. Mice were anesthetized with isoflurane or ketamine and instilled IN with silica or phosphate-buffered saline (PBS). Mice were sacrificed and lavaged after 3 days. To assess inflammation, alveolar cells were assessed by cytospin and lavage fluid was analyzed for inflammatory cytokines and total protein. While all parameters were increased in silica exposed groups, regardless of anesthesia type, there were significant increases in neutrophils and total protein in mice anesthetized with ketamine, compared to isoflurane. In comparing instillation techniques, mice were anesthetized with isoflurane and instilled IN, IT, or TO with silica. Increases were observed in all parameters, except tumor necrosis factor-alpha, following IT silica instillation as compared to the IN and TO instillation groups. In addition, fluorescent microsphere uptake by alveolar macrophages supported the notion that all methods of instillation were uniform, but IT had significantly greater dispersion. Taken together, these data show that each method of exposure tested generated significant inflammation among the silica groups, and any differences in parameters or techniques should be taken into consideration when developing an animal model to study pulmonary diseases. PMID- 20017596 TI - Hippocampus lipid peroxidation induced by residual oil fly ash intranasal instillation versus habituation to the open field. AB - Epidemiological studies have demonstrated the adverse effects of particulate matter (PM) inhalation on the respiratory and cardiovascular systems. It has been reported that air pollution may affect the central nervous system and decrease cognitive function. In rats, residual oil fly ash (ROFA) instillation causes decreased motor activity and increased lipid peroxidation in the striatum and the cerebellum. Our objective was to determine whether chronic instillation of particles induces changes in learning and memory in rats and whether oxidants in the hippocampus may contribute to these adverse effects. Forty-five-day-old male Wistar rats were exposed to ROFA by intranasal instillation and were treated with N-acetylcysteine (NAC) at 150 mg/kg i.p. for 30 days. Control groups were exposed to ROFA, NAC, or neither. On days 1, 8, and 30 of the protocol, rats were submitted to the open field test to evaluate habituation. After the last open field session, the rats were killed by decapitation. The hippocampus was used to determine lipid peroxidation (LP) by the thiobarbituric acid-reactive substances test. ROFA instillation induced an increase in LP in the hippocampus compared to all treatment groups (p = .012). NAC treatment blocked these changes. All of the treatment groups presented a decrease in the frequency of peripheral walking (p = .001), rearing (p = .001), and exploration (p = .001) over time. Our study demonstrates that exposure to particles for 30 days and/or NAC treatment do not modify habituation to an open field, a simple form of learning and memory in rats, and that oxidative damage induced by ROFA does not modulate these processes. PMID- 20017598 TI - Primary antifungal prophylaxis in acute myeloblastic leukemia and myelodysplastic syndrome--still an open question? AB - In this review, we aim to compare different early treatment strategies of invasive fungal diseases in patients undergoing induction chemotherapy for acute myelogenous leukemia or myelodysplastic syndrome. Three treatment approaches- prophylactic, empiric, and preemptive treatment--are subject to continuous discussion among physicians treating patients at risk. Considering the available clinical basis of evidence, we opt for antifungal prophylaxis with posaconazole 200 mg tid po as our primary prophylactic strategy, while the employment of preemptive treatment should be delayed until more accurate diagnostic tools become available. In addition to antifungal prophylaxis, empiric treatment with caspofungin or L-AmB may be administered to patients with fever resistant to broad-spectrum antibiotic treatment and without radiographic findings typical of invasive fungal disease. PMID- 20017600 TI - Improved survival using an intensive, pediatric-based chemotherapy regimen in adults with T-cell acute lymphoblastic leukemia. AB - All patients with newly diagnosed T-cell acute lymphoblastic leukemia (T-ALL) and treated over a 17-year period at a single institution were retrospectively analyzed. From 1990 to 2000, 40 patients were treated with a variety of adult based ALL regimens. From 2000 to 2007, a pediatric-based protocol, DFCI (Dana Farber Cancer Institute), was used as the standard regimen for all patients (n = 32). The two groups (DFCI and non-DFCI) had comparable baseline characteristics. Complete response rates were not significantly different between the DFCI- and non-DFCI-treated groups. The 3-year relapse free survival (RFS) and overall survival (OS) were significantly higher in the DFCI-treated group (p < 0.0001 and p = 0.0003, respectively). On multivariate analysis, the treatment group (DFCI vs. non-DFCI) was the major prognostic factor influencing both RFS and OS. The results provide evidence supporting the superior efficacy of asparaginase intensive pediatric-based regimens for adults with T-ALL. PMID- 20017599 TI - Report of a phase 1/2 study of a combination of azacitidine and cytarabine in acute myelogenous leukemia and high-risk myelodysplastic syndromes. AB - Cytarabine resistance characterizes relapsed and refractory acute myelogenous leukemia (AML). Restoration of cytarabine sensitivity can potentially improve treatment outcome in this setting. Acquired hypermethylation of gene promoters and associated silencing of gene expression has been implicated in chemo resistance, and drug-induced hypomethylation can improve sensitivity to cytarabine in vitro. We conducted an adaptively randomized study of a combination of azacitidine, a hypomethylating agent, and cytarabine in 34 patients with AML. The combination administered in a concomitant fashion is safe at full doses of azacitidine and cytarabine, without unexpected toxicities. However, in this advanced AML population, it was difficult to deliver more than one cycle of therapy, and minimal anti-leukemia activity was seen in patients with relapsed/refractory disease. Complete remission was achieved in 2 of 6 minimally pre-treated patients. We conclude that the combination of azacitidine and cytarabine is feasible but has limited activity in relapsed/refractory AML. PMID- 20017601 TI - Pharmaceutical development of microbicide drug products. AB - HIV infection rates in the developing world remain a serious problem. One potential approach to reduce infection rates is to use products known as microbicides, referred to herein as microbicide drug products (MDPs). These are drugs capable of, when administered topically to the vagina (or rectum), interfering with infection by one or more mechanisms. This review article covers the latest pharmaceutical developments in the area of microbicides dosage forms and delivery systems. These products are principally designed for use in the developing world and must therefore address cultural and societal issues generally unknown in the developed world. The first-generation microbicides evaluated clinically were principally polyanions. These drugs, administered intravaginally as gels, were found to be ineffective in preventing transmission of HIV from men to women. Second-generation drugs such as tenofovir, dapivirine, and UC781 are reverse transcriptase inhibitors developed as gels formulations and intravaginal rings (IVRs). Gels are considered coitally-related products while IVRs are coitally-independent systems designed to release the drug over a four week period or possibly longer (up to 3 or 4 months). Other dosage forms under development include fast dissolving films, tablets/capsules, and possibly vaginal sponges. Dual protection systems are also under development. These systems include formulations capable of preventing HIV infection along with a second drug capable of preventing conception or other viral infections such as HSV. PMID- 20017602 TI - Extracellular/microenvironmental redox state. AB - Extracellular redox (reduction-oxidation) state is a factor that serves as an important regulator of cell-microenvironmental interactions and is determined by several known variables; including redox-modulating proteins that are located on the plasma membrane or outside of cells, extracellular thiol/disulfide couples, and reactive oxygen species (ROS)/reactive nitrogen species (RNS) that are capable of traveling across plasma membranes into the extracellular space. The extracellular redox state works in concert with the intracellular redox state to control both the influx and efflux of ROS/RNS that may serve to modulate redox signaling or to perturb normal cellular processes or both. Under physiologic conditions, the extracellular space is known to have a relatively more-oxidized redox state than the interior of the cell. During pathologic conditions, such as cancer, the extracellular redox state may be altered, causing specific proteins such as proteases, soluble factors, or the extracellular matrix to have altered functions or activities. Recent studies have strongly supported an important relation between the extracellular redox state and cancer cell aggressiveness. The purpose of this review is to identify redox buffer networks in extracellular spaces and to emphasize the possible roles of the extracellular redox state in cancer, knowledge that may contribute to potential therapeutic interventions. PMID- 20017603 TI - Effect of ozone/oxygen mixture on systemic oxidative stress and organic damage. AB - Ozone is a molecule of high energetic content. Its great oxidative power has been used in medicine for the treatment of several illnesses with a wide spectrum. The rectal insufflation with a mixture of ozone/oxygen is considered as a simple therapy, not painful, of low cost and practically free from adverse effects. Given its potential oxidation and lack of side-effects, the objective has been to know the state of different indexes of redox state in blood which may contribute to understanding the mechanism by which mixtures of ozone/oxygen administered by intrarectal route are able to exert actions on other organs. With this purpose female rabbits were used, distributed into four groups, and three doses of ozone/oxygen mixture were tested. When treatment was finished, the determination of pro-oxidant and antioxidant markers was carried out. Also indexes of organic damage were determined. Ozone doses administered to rabbits did not cause adverse effects and mortality did not show significant changes relative to tissue damages and they increased enzymes activities belonging to the first line antioxidant defences. The results demonstrate that ozone/oxygen mixture administered by rectal insufflations is innocuous and it is able to increase the antioxidant defense of the organism. PMID- 20017604 TI - Cis-resveratrol glucuronidation kinetics in human and recombinant UGT1A sources. AB - It was hypothesized that cis-resveratrol glucuronidation contributes to a greater extent to in-vitro disposition of total resveratrol than previously assumed. To this end, the kinetic data for cis-resveratrol glucuronidation are reported. Glucuronidation assays were conducted in human liver and intestinal microsomes and in uridine diphosphate-glucuronosyltransferases (UGTs) UGT1A1, UGT1A6, UGT1A9, and UGT1A10. Kinetic parameters were estimated for the major cis resveratrol-3-O-glucuronide (cis-R3G). Substrate inhibition was observed with apparent V(max), K(m) and K(i) of 6.1 +/- 0.3/27.2 +/- 1.2 nmol min(-1) mg(-1), 415 +/- 48.1/989.9 +/- 92.8 and 789.6 +/- 76.3/1012 +/- 55.9 microM in human intestinal microsomes (HIMs) and UGT1A6, respectively (estimate +/- standard error (SE)). Biphasic kinetics were observed in human liver microsomes (HLMs), while sigmoidal kinetics were seen in UGT1A9 (V(max) = 11.92 +/- 0.2 nmol min(-1) mg(-1); K(m) = 360 microM; n = 1.27 +/- 0.07). The 4'-O-glucuronide (cis-R4'G) exhibited atypical kinetics in HLM, HIM, UGT1A1, and UGT1A10. UGT1A9 catalysed cis-R4'G formation at high substrate concentrations (V(max) = 0.33 +/- 0.015 nmol min(-1) mg(-1); K(m) = 537.8 +/- 67.8 microM). In conclusion, although the rates of formation of cis-R3G in HLM and UGT1A9 were higher than those for trans-R3G, the contribution to total resveratrol disposition could not be determined fully due to atypical kinetics observed. PMID- 20017605 TI - State-of-the-art in the management of chronic myelogenous leukemia in the era of the tyrosine kinase inhibitors: evolutionary trends in diagnosis, monitoring and treatment. AB - The treatment of patients with chronic myeloid leukemia (CML) continues to evolve rapidly as we gain better insights into the best monitoring strategies and as there is experience with the second generation tyrosine kinase inhibitors (TKI). Certain observations about CML and its clinical course remain relevant, such at its triphasic course and the prognostic value of the Sokal and Hasford scores. Other aspects of the disease including the most appropriate clinical monitoring and follow-up strategies and indications for changing therapy are evolving more rapidly. Best practice recommendations for monitoring of response have not only evolved over time but also affected by the availability and reliability of standard cytogenetics, FISH and molecular monitoring. Standard dose imatinib remains the best first-line therapy for most patients with first chronic phase CML. Patient and disease-related factors to evaluate when considering alternatives such as higher doses of imatinib, dasatinib, nilotinib and allogeneic transplant are discussed. PMID- 20017606 TI - Treatment of Philadelphia chromosome-positive acute lymphoblastic leukemia. AB - The presence of the Philadelphia chromosome is a poor prognosis factor in acute lymphoblastic leukemia (ALL), in both children and adults. Using molecular techniques of the gen bcr/abl, it is possible to detect the abnormality, in up to the 40% of adult patients. The unsatisfactory results with conventional chemotherapy schemes have determined the intensification of the treatments and the consideration of allogenic bone marrow transplants as the best therapeutic instance. The development of tyrosine kinase inhibitors have become a therapeutic improvement in the treatment of Philadelphia chromosome-positive ALL, being combined with chemotherapy schemes, only in a selected group of patients, even in therapeutic programs that include transplant. PMID- 20017607 TI - Advances in treatment of chronic myelogenous leukemia--new treatment options with tyrosine kinase inhibitors. AB - Imatinib is considered standard therapy for patients with chronic myelogenous leukemia (CML), inducing a high rate of hematologic and cytogenetic responses. Despite these excellent results, several patients develop resistance to imatinib. Mechanisms of resistance are varied and include BCR-ABL1 kinase domain mutations, decreased entry of imatinib into cells, acquisition of secondary genetic changes and activation of alternate signaling pathways. Second-generation tyrosine kinase inhibitors (TKI) (dasatinib, nilotinib) were developed as an alternative for patients that develop resistance or are intolerant to imatinib. Dasatinib is a dual Abl/Src kinase TKI that is structurally unrelated to imatinib and is approved for therapy of all phases of CML in patients who are resistant or intolerant to imatinib. Nilotinib is a compound related to imatinib that has greater specificity and improved binding characteristics, and has clinical activity in the setting of imatinib failure. Resistance to multiple TKIs does occur, particularly in patients with the T315I mutation. Several new agents are in development including new TKIs, aurora kinase inhibitors and homoharringtonine. PMID- 20017608 TI - Hematopoietic stem cell transplantation in the era of tyrosine kinase inhibitors. AB - Until the 1990 s, the treatment of chronic myeloid leukemia (CML) recognized hematopoietic stem cell transplantation as the best treatment for those patients with an available donor. With the advent of imatinib in 2001, this paradigm changed dramatically as this drug provided outstanding and durable rates of hematologic, cytogenetic, and molecular responses. As a consequence it became the gold standard first-line treatment for most patients. However, after almost a decade of its use, it is clear that although very effective, imatinib cannot cure CML as transplantation has already proven so. Furthermore, the new non myeloablative regimens and the improvements in survival after allogeneic transplant, especially in the field of unrelated transplants, offer this option to a broader population of patients with CML. This adds to the old question of whom to transplant, when and how to proceed with the allograft. This article reviews the current role of transplantation in the era of tyrosine kinase inhibitors and will try to elucidate its role in the frontline setting as well as after first- and second-line kinase inhibitors failure. PMID- 20017609 TI - Pharmacological inhibition of phosphoinositide 3 and TOR kinases improves survival of Drosophila melanogaster. AB - Recent progress in our understanding of genetic mechanisms of aging and longevity provides an opportunity to select some enzymes as targets for pharmacological correction. The phosphoinositide 3-kinase (PI3K) and TOR-kinase cascades are affected in some long-lived mutants of different animals, such as nematodes and mice. The purpose of this study was to investigate the geroprotector efficiency of the inhibitors of enzymes that are known to be affected in long-lived mutants. Experimental animals were exposed to low dozes of LY-294002 (5 microM), wortmannin (0.5 microM), and rapamycin (0.5 microM) separately during their lifetimes. We have shown that the specific PI3K inhibitors (LY-294002 and wortmannin) and the TOR-kinase inhibitor rapamycin slightly increase the median and maximal lifespan of the fruit fly, Drosophila melanogaster. PMID- 20017610 TI - Regression analysis of walking parameters for the age-predictive equation. AB - We tried to establish an age-predictive equation from kinematic and kinetic parameters during walking using the 2007 Okada database of Japanese elderly. Some predictive equations were established for each gender (109 males and 139 females) -once for all variables, second for the stepwise variable selection method. The range of motion (ROM) of knee and hip and step length are the stable parameters in age-predictive models for both genders. Although the model achieved the Minimum Akaike's Information Criterion Estimate (MAICE), it did not reflect sufficiently the regimes of variable selection. PMID- 20017611 TI - Carnosine as a natural antioxidant and geroprotector: from molecular mechanisms to clinical trials. AB - Carnosine is a neuroprotective dipeptide consisting of beta-alanine and L histidine. It demonstrates a number of useful features, including stimulation of brain and muscle microcirculation and a rejuvenating effect on cultured cells. Its activity is based on its antioxidant and antiglycating action that, in addition to heavy metal chelation and pH-buffering ability, makes carnosine an essential factor for preventing neurodegeneration and accumulation of senile features. Recently, carnosine was successfully used to treat patients after brain stroke or patients with Parkinson disease. We conclude that carnosine can be recommended for patients under oxidative stress as a natural remedy having high efficiency and no side effects. PMID- 20017612 TI - Amyloid formation: age-related mechanism in Creutzfeldt-Jakob disease? AB - Protein aggregation occurs in many age-related neurodegenerative diseases, where it can lead to deposits of naturally occurring proteins in the brain. In case of Creutzfeldt-Jakob disease (CJD), these deposits consist of prion protein (PrP). CJD has three etiologies: spontaneous, genetic, or caused by infection. A polymorphism within the PrP gene is associated with susceptibility of infection. The main event in prion diseases is the conversion of PrP from its naturally occurring isoform to its disease-associated isoform. Here, we present the adaption of a previously reported in vitro conversion system based on hamster recombinant PrP to analyze amyloid fibril formation of human recombinant PrP. We further compare the aggregation characteristics of the human PrP according to the polymorphism variants M129 and V129. PMID- 20017613 TI - Exercise intensity and antioxidant ability. AB - We performed maximal exercise tests on 6 men using a bicycle ergometer and then measured the ferric-reducing ability in plasma to clarify the relationship between antioxidant status and exercise intensity. The ferric-reducing ability showed a rectilinear increase with the increase in exercise intensity, whereas the generation of hydroxyl radicals showed a decrease with the increase in exercise intensity. Also, the generation of hydroxyl radicals showed a symmetrical correlation to lactate level in plasma. These results suggest that antioxidant ability is enhanced with an increase in exercise intensity, and that the lactate level, which increases with physical exercise, may act as a scavenger of free radicals. PMID- 20017616 TI - Papillary thyroid carcinoma mimicking anaplastic thyroid carcinoma following fine needle aspiration biopsy. AB - BACKGROUND: Cutaneous metastases from well-differentiated thyroid carcinomas are rare and usually identified in patients with widely disseminated disease. Fine needle aspiration biopsy (FNAB) has become an acceptable method for the assessment of thyroid nodules. Very rarely needle track dissemination of tumor cells in the thyroid nodule occurs, but, when this occurs, it is evident many years after the FNAB. We report a patient who appears to have tumor dissemination from an FNAB needle track only 4 months after the procedure. SUMMARY: An 85-year old female presented with a mass on her neck, skin ulceration, and hemorrhage 4 months after FNAB was performed for a thyroid nodule by another physician. A second FNAB with ultrasound guidance yielded cytology diagnostic of papillary thyroid carcinoma. Papillary thyroid carcinoma was confirmed by surgical dissection of the mass, and a linear array of tumor was noted in skin and muscle was performed again, and the cytological diagnosis revealed papillary carcinoma. After surgical resection, the histopathological diagnosis determined the nodules to be papillary carcinoma. Metastatic deposits appeared in the skin and the muscle. The linear array and the site of metastases implied that seeding most probably resulted from the needle biopsy. CONCLUSION: Although FNAB is a useful tool for the diagnosis of thyroid nodules, it is important to consider the risk of tumor cell dissemination. PMID- 20017617 TI - Does recombinant human thyrotropin-stimulated positron emission tomography with [18F]fluoro-2-deoxy-D-glucose improve detection of recurrence of well differentiated thyroid carcinoma in patients with low serum thyroglobulin? AB - BACKGROUND: Thyrotropin (TSH) stimulates thyrocyte metabolism, glucose transport, and glycolysis. The interest in using recombinant human TSH (rhTSH) stimulation of fluoro-2-deoxy-D-glucose (FDG) with positron emission tomography (PET) has been shown, but mainly for patients with high serum thyroglobulin (Tg) concentration. We evaluated the use of rhTSH-stimulated PET-FDG in patients with low serum Tg concentration. METHODS: Sixty-one PET/computed tomography (CT)-FDG (Biograph Sensation 16; Siemens Medical Solutions, Knoxville, TN) were performed in 44 patients (28 women and 16 men; 51 +/- 16 years) with positive Tg levels, negative or no contributive iodine-131 whole-body scintigraphy results, and no contributive morphological imaging results (ultrasound, magnetic resonance imaging, and CT). Thirty-eight patients had papillary carcinoma and six had follicular thyroid carcinoma. All patients had previously undergone total thyroidectomy and postoperative iodine ablation of thyroid bed remnant tissue. The rhTSH-stimulated PET/CT-FDG (5 MBq/kg) was performed after two 0.9 mg intramuscular doses of rhTSH (Thyrogen; Genzyme) which were administered 48 and 24 hours before imaging, while patients continued levothyroxine (LT(4)). Blood sampling was performed immediately before FDG injection for measurement of serum TSH and Tg concentrations (TSH(1) and Tg(1)) and after 48 hours (TSH(2) and Tg(2)). PET/CT-FDG findings were compared with the Tg: (i) at the initial iodine treatment during T(4) withdrawal (Tg(ini)), (ii) under T(4) (Tg(T4)) within 3 months before the PET/CT-FDG, (iii) with Tg(1), and (iv) with Tg(2). PET/CT-FDG findings were correlated with the findings of histology, iodine-131 whole-body scintigraphy, morphological imaging, or clinical follow-up. RESULTS: The mean Tg(ini) was 785 +/- 2707 microg/L for a TSH of 73 +/- 64 mU/L. The mean Tg(T4) was 7 +/- 15 microg/L (T(4) = 195 +/- 59 microg/day; mean TSH of 0.24 +/- 0.57 mU/L). Among the 44 patients, PET/CT-FDG findings were positive in 20 and negative in 24. Among the 61 PET/CT-FDG, 25 PET/CT-FDG were positive (41%). Among the 25 positive PET, the Tg(T4) values were less than 10 microg/L for 19, including 9 true-positive patients (20% of the 44 patients). There was no difference of PET/CT-FDG results (positive vs. negative) as related to the serum Tg concentrations (p = 0.99 for Tg(ini), p = 0.95 for Tg(T4), p = 0.07 for Tg(1), and p = 0.42 for Tg(2)). No relation was observed with PET/CT-FDG results and initial tumor size (p = 0.52) or node metastasis (p = 0.14). CONCLUSION: In the diagnosis of recurrent disease in patients with differentiated thyroid carcinoma and low Tg level, the sensitivity of rhTSH-stimulated PET/CT-FDG seems to be low and no correlation was observed between PET/CT-FDG findings and Tg level. However, positive PET-FDG results have been found in 9/44 (20%) patients with serum Tg levels lower than 10 microg/L. Therefore, this series shows that a cutoff value of 10 microg/L for the Tg under T(4) is probably not the best criteria to select patient candidates for PET/CT-FDG examination to detect the recurrence of differentiated thyroid carcinoma. PMID- 20017618 TI - Bilateral lymphoepithelial cysts of the thyroid gland. PMID- 20017619 TI - Expressions of vascular endothelial growth factor and nitric oxide synthase III in the thyroid gland of ovariectomized rats are upregulated by estrogen and selective estrogen receptor modulators. AB - BACKGROUND: Estrogen promotes the growth of thyroid cells. Therefore, we analyzed the influence of estrogen and selective estrogen receptor modulators (SERMs) on the expression of vascular endothelial growth factor (VEGF) and nitric oxide synthase III (NOS III) in the thyroid gland of ovariectomized (Ovx) rats. METHODS: Wistar rats were divided into five groups, and bilateral ovariectomies were performed, except on the Sham-operated controls (Sham). Rats were grouped as follows: Sham; Ovx; and Ovx rats treated with daily subcutaneous injections of estradiol benzoate 3.5 microg/kg, tamoxifen 2.5 mg/kg, or raloxifene 2.5 mg/kg for 50 consecutive days. Control animals received vehicle (propyleneglycol), and at the end of the treatment, rats were sacrificed. The thyroid glands were excised, weighed, and processed for analysis of the expression of VEGF or NOS III by immunohistochemistry. The mean vascular areas were evaluated by immunodetection of alpha-smooth muscle actin. RESULTS: Thyroid weight and mean vascular area were lower in Ovx as compared with Sham, Ovx + estradiol benzoate, Ovx + Tam, or Ovx + Ral (p < 0.01). VEGF (p < 0.01) and NOS III expressions (p < 0.05) were significantly lower in the Ovx group, as compared with Sham, Ovx + estradiol benzoate, Ovx + Tam, and Ovx + Ral. Immunoreactivity for both VEGF and NOS III was mainly detected in the cytoplasm of the follicular epithelial cells. CONCLUSIONS: Our data suggest that estrogen and SERMs regulate the thyroid gland vascularization and that tamoxifen and raloxifene behave like estrogen does. Estrogen and SERMs upregulate VEGF and NOS III in such a way as to reverse the effects detected on the thyroid microvasculature of the Ovx rats. PMID- 20017620 TI - A stimulatory thyrotropin receptor antibody (M22) and thyrotropin increase interleukin-6 expression and secretion in Graves' orbital preadipocyte fibroblasts. AB - BACKGROUND: Patients with Graves' ophthalmopathy (GO) have circulating autoantibodies directed against the thyrotropin receptor (TSHR) and elevated levels of the proinflammatory cytokine interleukin-6 (IL-6) in both serum and orbital tissues. We hypothesized that these autoantibodies might increase IL-6 expression and secretion in preadipocyte fibroblasts and adipocytes from patients with GO, and thus directly impact the clinical activity of the disease. METHODS: IL-6 mRNA levels were measured in cultures of GO orbital preadipocytes (n = 3) treated during adipocyte differentiation with a monoclonal stimulatory TSHR antibody (M22; 10 ng/mL), IL-6 (1 ng/mL), or TSH (10 U/L). Additionally, levels of IL-6 protein secretion were assessed after adipocyte differentiation in orbital cultures exposed to TSH or M22 for 24 or 48 hours (n = 8). IL-6 mRNA levels were also measured in orbital adipose tissue specimens from well characterized GO patients (n = 9) and normal individuals (n = 9). RESULTS: Treatment of GO orbital preadipocyte cultures with IL-6, TSH, or M22 during adipocyte differentiation resulted in increased IL-6 mRNA levels (3.1-fold, 2.9 fold, and 2.7-fold, respectively; p < 0.05). Treatment of orbital cultures with M22 or TSH after adipocyte differentiation enhanced the release of IL-6 protein into the medium at both 24 and 48 hours for TSH (mean 1.9- and 2.3-fold; p = 0.002 and 0.015, respectively) and at 48 hours for M22 (mean 2.0-fold; p = 0.005). In addition, we found mean IL-6 mRNA levels to be significantly increased in GO orbital adipose tissue specimens (10-fold; p < 0.01), primarily attributable to high levels in three of the four patients with clinical activity scores >or=5. CONCLUSIONS: Both TSH and M22 increase IL-6 expression in orbital preadipocyte fibroblasts and IL-6 secretion by mature adipocytes. These results suggest that circulating TSHR autoantibodies in GO might play a direct role in the clinical activity of the disease. PMID- 20017624 TI - The non thermal effect of weak intensity millimeter waves on physicochemical properties of water and water solutions. AB - The comparative study of the effects of 5.8 mW/cm(2) Millimeter Waves (MMW) and near Infrared (IR) irradiation on thermal properties, specific adsorption rate (SAR), specific electrical conductivity (SEC) and hydrogen peroxide (H(2)O(2)) content of distilled water (DW), and physiological solutions (PS) was performed. The thermal effect of MMW irradiation appeared only after the first minute of irradiation, while the IR heating started from the first minute of irradiation. The heat fusion of frozen MMW-treated DW and PS was significantly less than sham and IR-treated DW and PS. MMW irradiation had time-dependent elevation effect on water SEC and SAR, which was accompanied by the increase of H(2)O(2) formation in it. We suggest that the MMW-induced vibration of water dipole molecules caused the non thermal changes of physicochemical properties of DW and PS, which promote the formation of H(2)O(2) in water. PMID- 20017625 TI - Effect of mobile phone exposure on apoptotic glial cells and status of oxidative stress in rat brain. AB - The aim of this study was to investigate the effects of mobile phone exposure on glial cells in brain. The study carried out on 31 Wistar Albino adult male rats. The rat heads in a carousel exposed to 900 MHz microwave. For the study group (n:14), rats exposed to the radiation 2 h per day (7 days in a week) for 10 months. For the sham group (n:7), rats were placed into the carousel and the same procedure was applied except that the generator was turned off. For the cage control (n:10), nothing applied to rats in this group. In this study, rats were euthanized after 10 months of exposure periods and brains were removed. Brain tissues were immunohistochemically stained for the active (cleaved) caspase-3, which is a well-known apoptosis marker, and p53. The expression of the proteins was evaluated by a semi-quantitative scoring system. However, total antioxidative capacity (TAC), catalase, total oxidant status (TOS), and oxidative stress index were measured in rat brain. Final score for apoptosis in the exposed group was significantly lower than the sham (p < 0.001) and the cage control groups (p < 0.01). p53 was not significantly changed by the exposure (p > 0.05). The total antioxidant capacity and catalase in the experimental group was found higher than that in the sham group (p < 0.001, p < 0.05). In terms of the TOS and oxidative stress index, there was no statistically significant difference between exposure and sham groups (p > 0.05). In conclusion, the final score for apoptosis, total antioxidant capacity and catalase in rat brain might be altered by 900 MHz radiation produced by a generator to represent exposure of global systems for mobile communication (GSM) cellular phones. PMID- 20017626 TI - Electrostatic contributions to colchicine binding within tubulin isotypes. AB - Tubulin, the structural subunit of microtubules, is the target of some highly successful anti-tumor drugs. Most of these drugs bind to the beta-tubulin resulting in the inhibition of microtubule dynamics and eventually cell death. The varied cellular distribution of several human isotypes of beta -tubulin provides a platform upon which to construct novel chemotherapeutic agents that are able to differentiate between these types of cells. To test this hypothesis, we have previously created homology models of the nine most frequently observed human beta -tubulin isotypes and analyzed them for differences in the colchicine binding site. Here, we describe the electrostatic properties of the colchicine binding site and how this may affect calculated drug binding affinities between the beta -tubulin isotypes. PMID- 20017627 TI - Specific patterns of weak (1 microTesla) transcerebral complex magnetic fields differentially affect depression, fatigue, and confusion in normal volunteers. AB - Normal young adults were exposed for 20 min once per week for a total of 3 sessions to 1 of 7 configurations of weak (1 microTesla) magnetic fields or to a sham field. The fields were spatially rotated and applied through the brain at the level of the temporoparietal lobes. The Profile of Mood States was taken before and after each session. Before, during, and after the treatments, heart rate, plethysmographic activity, and skin conductance were measured by computer. The results indicated that the burst-firing pattern previously demonstrated to be effective for clinical depression, improved mood and vigour compared to the sham field or other treatments. Subjects who were exposed to a burst-firing pattern, a complex-sequenced pattern, and a pattern whose electrical equivalents stimulate long-term potential in hippocampus slices also exhibited less psychometric fatigue after the sessions compared to subjects who received the sham field or random-sequenced fields. These results replicate previous studies and indicate that rationally designed complex patterns of magnetic fields may simulate pharmacological treatments. PMID- 20017628 TI - Entrainment of stage 2 sleep spindles by weak, transcerebral magnetic stimulation in an "epileptic" woman. AB - The proportions of Stage 2 spindles present per second over the prefrontal and temporal lobes of a patient who had been experiencing complex partial epileptic like experiences and disturbances in sleep patterns for four years following a closed head injury were measured while a weak (1 microT) burst-firing magnetic field was applied transcerebrally through the temporoparietal regions. In several successive segments during the exposure the onsets of the spindles were significantly correlated (rhos between 0.30 and 0.40) with the onset of the field that was presented once every 3 s for about 0.7 s. Lag/lead analyses indicated only the contemporary occurrence of the fields and the spindles were significantly correlated over time. These results suggest that some states of consciousness, particularly those associated with the "epilepsy spectrum disorder" may be more sensitive to entrainment or "synchronization" by weak complex magnetic fields whose temporal structures are designed to simulate natural brain activity. PMID- 20017629 TI - Qualitative effect on mRNAs of injury-associated proteins by cell phone like radiation in rat facial nerves. AB - Rats were exposed to cell phone radiation for 6 hours per day for 18 weeks. The buccal and mandibular branches of the facial nerve were evaluated for this study. The mRNA levels of four proteins that are usually up regulated when an injury has occurred were investigated; included were Calcium ATP-ase, Endothelin, Neural Cell Adhesion Molecule, and Neural Growth Factor. These isolated mRNAs were subjected to RT-PCR and all four were up regulated. The mandibular nerve showed a higher and broader level of up regulation than the buccal nerve. All four mRNA up regulations for the mandibular nerve and two for the buccal nerve were also statistically significant. These specific injury-related findings were mild. As the use of these cell phones continues, there most likely will be permanent damage to these tissues over the years and the likelihood of tumors, cancers, and system failures will potentially increase. PMID- 20017630 TI - Histopathological and ultrastructural studies on the effects of electromagnetic fields on the liver of preincubated white Leghorn chicken embryo. AB - There are several reports indicating a linkage between exposures to 50-60 Hz electromagnetic fields and abnormalities in the early stages of embryonic development of chicken embryos. The present study was designed to demonstrate whether electromagnetic fields could be an environmental factor invoking histopathological and ultra-structural changes in livers of preincubated chicken embryos exposed to EMFs. Following other researchers and our previous results from different groups of Developmental Biology at the Animal Sciences, Faculty of Biological Sciences, Shahid-Beheshti University, effects of most effective intensities (1.33, 2.66, 5.52, and 7.32 mT) of electromagnetic fields (EMFs, 50 Hz ) on livers of pre-incubated white leghorn chicken embryos were investigated . 150 healthy, fresh, and fertilized eggs (55-65 gr) were divided into 6 groups of experimental(1-4, n = 30), control (n = 60), and sham (n = 50). Experimental eggs (inside coil) were exposed to 4 different intensities (1.33, 2.66, 5.52, and 7.32 mT). Sham groups were located inside same coil, with no exposure, for 24 h before incubation. Control, sham, and experimental groups (1-4) were then incubated in an incubator (38 +/- 0.5 degrees C, 60% humidity) for 17 days. At the end of this period, livers of experimental, sham, and control groups were processed for light and transmission electrom microscopes (TEM and SEM) studies. So, livers of 17-day old chicken embryos were removed by C-sections, fixed in formalin 10%, stained with H&E and reticulin, and studied under light microscope. Others were prepared for electron microscopes (TEM and SEM) investigations. Morphological observations indicated exencephalic embryos, embryos with asymmetrical faces, crossed beak, shorter upper beak, deformed hind limbs, gastroschesis, anophthalmia, and microphthalmia. H&E and reticulin stainings, TEMS, and SEMs studies indicated EMFs would create hepato-cytes with fibrotic bands, severe steatohepatitis, vacuolizations, swollen and extremely electron-dense mitochondria, reduced invisible cristae, crystalized mitochondria with degenerated cristae, myelin-like figures, macrophages engulfing adjacent cells, dentated nuclei, nuclei with irregular envelopes, degenerated hepatocytes, abnormal lipid accumulations, lipid droplets pushing hepatocytes' nuclei to the corner of the cells, abundant cellular infiltrations cellular infiltrations inside sinusoid and around central veins, disrupted reticulin plexus, and release of chromatin into cytosol,, with partially regular water layers. An elevated oxyradical generation and, subsequently, cell membrane disruptions were the reasons for electromagnetic fields inducing cell damages. PMID- 20017631 TI - Preterm infants' T cell responses to inactivated poliovirus vaccine. AB - BACKGROUND: The antigen-specific T cell responses of preterm infants to immunization are not well understood. The aim of the present study was to compare the T cell responses of preterm infants after inactivated poliovirus vaccination with those of term infants. METHODS: We prospectively enrolled 2-month-old preterm (gestational age, 33 weeks) and term (gestational age, 37 weeks) infants to receive 3 doses of diphtheria-tetanus toxoids-acellular pertussis-hepatitis B virus-inactivated poliovirus vaccine. Whole blood and peripheral blood mononuclear cells (PBMCs) were stimulated with poliovirus vaccine, and memory T cell activation was analyzed by flow cytometry and lymphoproliferation, respectively. Levels of poliovirus neutralizing antibodies were measured in serum. RESULTS: We enrolled 33 preterm and 50 term infants. Preterm infants had fewer circulating CD4(+)CD45RO(+) memory (P = .005) and CD4(+)CD69(+)IFN-gamma(+) cells activated by staphylococcus enterotoxin B at 2 (P = .015) and 7 (P = .05) months of age. After immunization, preterm and term infants had comparable frequencies of poliovirus-specific CD4(+)CD45RO(+)CD69(+)IFN-gamma(+) memory T cells (P = .79). PBMCs from preterm infants had diminished poliovirus-specific lymphoproliferation (P<.001). Although all infants developed seroprotective poliovirus antibody titers, serotype 1 titers were lower among preterm infants (P = .03). CONCLUSIONS: Preterm infants develop poliovirus-specific T cell responses that are comparable to those of term infants. However, they demonstrate nonspecific and poliovirus-specific functional T cell limitations, suggesting that investigations into whether T cell differences remain as preterm infants mature are warranted. PMID- 20017632 TI - Enhanced memory responses to seasonal H1N1 influenza vaccination of the skin with the use of vaccine-coated microneedles. AB - BACKGROUND: Morbidity and mortality due to influenza could be reduced by improved vaccination. METHODS: To develop a novel skin delivery method that is simple and allows for easy self-administration, we prepared microneedle patches with stabilized influenza vaccine and investigated their protective immune responses. RESULTS: Mice vaccinated with a single microneedle dose of trehalose-stabilized influenza vaccine developed strong antibody responses that were long-lived. Compared with traditional intramuscular vaccination, stabilized microneedle vaccination was superior in inducing protective immunity, as was evidenced by efficient clearance of virus from the lung and enhanced humoral and antibody secreting cell immune responses after 100% survival from lethal challenge. Vaccine stabilization was found to be important, because mice vaccinated with an unstabilized microneedle vaccine elicited a weaker immunoglobulin G 2a antibody response, compared with the stabilized microneedle vaccine, and were only partially protected against viral challenge. Improved trafficking of dendritic cells to regional lymph nodes as a result of microneedle delivery to the skin might play a role in contributing to improved protective immunity. CONCLUSIONS: These findings suggest that vaccination of the skin using a microneedle patch can improve protective efficacy and induce long-term sustained immunogenicity and may also provide a simple method of administration to improve influenza vaccination coverage. PMID- 20017634 TI - Influenza vaccination rates among healthcare workers in the department of veterans affairs community living centers. PMID- 20017633 TI - Successful implementation of the World Health Organization hand hygiene improvement strategy in a referral hospital in Mali, Africa. AB - OBJECTIVE: To assess the feasibility and effectiveness of the World Health Organization hand hygiene improvement strategy in a low-income African country. DESIGN: A before-and-after study from December 2006 through June 2008, with a 6 month baseline evaluation period and a follow-up period of 8 months from the beginning of the intervention. SETTING: University Hospital, Bamako, Mali. Participants. Two hundred twenty-four healthcare workers. METHODS: The intervention consisted of introducing a locally produced, alcohol-based handrub; monitoring hand hygiene compliance; providing performance feedback; educating staff; posting reminders in the workplace; and promoting an institutional safety climate according to the World Health Organization multimodal hand hygiene improvement strategy. Hand hygiene infrastructure, compliance, healthcare workers' knowledge and perceptions, and handrub consumption were evaluated at baseline and at follow-up. RESULTS: Severe deficiencies in the infrastructure for hand hygiene were identified before the intervention. Local handrub production and quality control proved to be feasible, affordable, and satisfactory. At follow-up, handrubbing was the quasi-exclusive hand hygiene technique (93.3%). Compliance increased from 8.0% at baseline to 21.8% at follow-up (P < .001). Improvement was observed across all professional categories and medical specialities and was independently associated with the intervention (odds ratio, 2.50; 95% confidence interval, 1.8-3.5). Knowledge enhanced significantly (P < .05), and perception surveys showed a high appreciation of each strategy component by staff. CONCLUSIONS: Multimodal hand hygiene promotion is feasible and effective in a low-income country. Access to handrub was critical for its success. These findings motivated the government of Mali to expand the intervention nationwide. This experience represents a significant advancement for patient safety in developing countries. PMID- 20017635 TI - Focus group study of hand hygiene practice among healthcare workers in a teaching hospital in Toronto, Canada. AB - OBJECTIVE: To understand the behavioral determinants of hand hygiene in our hospital. DESIGN: Qualitative study based on 17 focus groups. SETTING: Mount Sinai Hospital, an acute care tertiary hospital affiliated with the University of Toronto. PARTICIPANTS: We recruited 153 healthcare workers (HCWs) representing all major patient care job categories. METHODS: Focus group discussions were transcribed verbatim. Thematic analysis was independently conducted by 3 investigators. RESULTS: Participants reported that the realities of their workload (eg, urgent care and interruptions) make complete adherence to hand hygiene impossible. The guidelines were described as overly conservative, and participants expressed that their judgement is adequate to determine when to perform hand hygiene. Discussions revealed gaps in knowledge among participants; most participants expressed interest in more information and education. Participants reported self-protection as the primary reason for the performance of hand hygiene, and many admitted to prolonged glove use because it gave them a sense of protection. Limited access to hand hygiene products was a source of frustration, as was confusion related to hospital equipment as potential vehicles for transmission of infection. Participants said that they noticed other HCWs' adherence and reported that others HCWs' hygiene practices influenced their own attitudes and practices. In particular, HCWs perceive physicians as role models; physicians, however, do not see themselves as such. CONCLUSIONS: Our results confirm previous findings that hand hygiene is practiced for personal protection, that limited access to supplies is a barrier, and that role models and a sense of team effort encourage hand hygiene. Educating HCWs on how to manage workload with guideline adherence and addressing contaminated hospital equipment may improve compliance. PMID- 20017636 TI - Doing good and doing it well, especially where it is not easy. PMID- 20017637 TI - Predictors of virologic failure in HIV-1-infected adults receiving first-line antiretroviral therapy in 8 provinces in China. AB - BACKGROUND: Despite poor primary health care systems, free antiretroviral therapy (ART) has been available in China for >5 years. Virologic outcomes in Chinese patients receiving ART have not been described on a national level. METHODS: A multistage cluster design was used in 8 provinces to randomly select patients who had been receiving first-line ART for at least 6 months, who were stratified into 3 treatment-duration groups. Viral load testing and patient interviews were conducted, and data were linked with national treatment database information. Collected data were analyzed for association with viral suppression by means of multivariate modeling. Adequate viral suppression was defined as a viral load of <400 copies/mL. RESULTS: Of 5256 patients receiving ART, 3894 met the eligibility criteria, among whom 1153 were analyzed. Overall, 72% demonstrated viral suppression, and 82%, 73%, and 67% of the participants receiving ART for 6-11, 12 23, and > or = 24 months, respectively, experienced viral suppression (P < .001). In a multivariate model, treatment given at locations other than county-level hospitals was less likely to achieve viral suppression, with greater odds for inadequate virologic response found at village clinics (odds ratio [OR], 5.4; 95% confidence interval [CI], 2.9-10.1), township health centers (OR, 3.1; 95% CI, 1.7-5.6), and public health clinics (OR, 3.1; 95% CI, 1.7-5.6). Patients receiving didanosine-based regimens were more likely to experience an inadequate virologic response than were those receiving lamivudine-based regimens (OR, 3.9; 95% CI, 2.7-5.7). CONCLUSIONS: China's national ART program is largely successful at suppressing viral load. Care received outside of hospitals and regimens containing didanosine were associated with less favorable virologic outcomes. PMID- 20017638 TI - Syncytial knots as a reflection of placental maturity: reference values for 20 to 40 weeks' gestational age. AB - Syncytiotrophoblastic knots or syncytial knots are aggregates of syncytial nuclei at the surface of terminal villi. In the term placenta, most syncytial knots are thought to be artifacts from tangential sectioning while the minority are syncytial sprouts, bridges, or apoptotic knots. Syncytial knots are consistently present, increasing with increasing gestational age, and can be used to evaluate villous maturity. Increased syncytial knots are associated with conditions of uteroplacental malperfusion and are important in placental examination. Although 30% of terminal villi with syncytial knots at term are often reported, no reference values have been developed for the percentage of villi with syncytial knots at different gestational ages. We counted the percentage of chorionic villi with syncytial knots at different gestational ages from 20 to 40 weeks using cases with no history of malperfusion or clinical conditions known to be associated with malperfusion. We provide normal reference data for the average percentage of syncytial knots for gestational ages ranging from 20 to 40 weeks. There was a significant positive correlation of gestational age with percentage of villi with syncytial knots. Term placentas (37-40 weeks) showed an average of 28% syncytial knots. A drop-off to a mean of 22.5% was noted at 36 weeks; at 26 to 33 weeks, syncytial knots varied from 10.8% to 14.7%; between 20 and 25 weeks, syncytial knots ranged between 5.2% and 9.l%. These reference data can facilitate histologic assessment of normal placental maturation as well as evaluation of placental morphology in placental malperfusion. PMID- 20017639 TI - NUT midline carcinoma in a newborn with multiorgan disseminated tumor and a 2 year-old with a pancreatic/hepatic primary. AB - NUT midline carcinoma (NMC) is a rare and aggressive malignant epithelial tumor defined by rearrangement of the NUT gene on chromosome 15. In two thirds of cases, NUT is involved in a balanced translocation with BDR4 on chromosome 19, while in the remaining cases, NUT is rearranged with variant fusion partners such as BRD3. These undifferentiated tumors primarily affect midline structures, usually in the upper aerodigestive tract and mediastinum. Most reported cases have followed a rapidly lethal clinical course. We report the clinical and pathological findings of NMC in the youngest patients identified so far. The 1st case involves a newborn who presented with a supraorbital mass and extensive multiorgan involvement, including the spine, lungs, liver, pancreas, adrenal glands, and subcutaneous tissue. The 2nd patient was a 2-year-old male with an abdominal mass involving the liver and pancreas with pulmonary metastasis. Histopathological analysis of both tumors showed undifferentiated malignant neoplasms, and immunohistochemistry showed positivity for epithelial markers. Both tumors demonstrated t(15;19), and immunohistochemistry with NUT monoclonal antibodies and fluorescent in situ hybridization confirmed NUT rearrangement. The patients died from disease at 1 and 2 months postpresentation. Thus far, 25 cases have been reported, including our 2 current cases. Presentation ages range from 0 to 78 years (mean, 23 years). Herein, we report the 2 youngest reported cases of NMC, including the 1st congenital case and the 1st case arising within the liver/pancreas. Increased awareness and further molecular studies are required for a better understanding of NMC pathobiology and improved therapeutic outcomes. PMID- 20017640 TI - Maternal floor infarction associated with oligohydramnios and cystic renal dysplasia: report of 2 cases. AB - Maternal floor infarction (MFI) is an unusual, idiopathic placental disorder characterized by deposition of amorphous fibrinoid material along the maternal aspect of the intervillous space. This condition is associated with poor perinatal outcome-in particular, spontaneous abortion-fetal growth restriction, and stillbirth, with a high recurrence rate in subsequent pregnancies. It is unknown whether MFI is a single entity or the common end point of different insults. Most studies have linked MFI to underlying maternal disorders including gestational hypertension, autoimmune disease, and thrombophilia. In contrast, there have been only a few case reports regarding the possibility of a fetal basis for MFI. We report 2 cases of MFI in fetuses who suffered from oligohydramnios as a result of bilateral cystic renal dysplasia. These 2 cases suggest the concept that fetoplacental factors may also play a role in MFI. It is speculated that the mechanism might involve changes in intrauterine hydrostatic pressure gradients. PMID- 20017642 TI - Force and movement of non-osseointegrated implants: an in vitro study. AB - Dental implants have enabled a dramatic increase in the quality of life for many partially edentulous and edentulous patents. Immediate loading of newly placed dental implants is a recent advancement that attempts to meet patient demand. However, immediate loading of a just placed implant may induce implant failure to osseointegrate. Some patients can generate a biting force that can reach approximately 1300 Newtons (N) in the posterior jaws. The magnitude of bite force that would cause failure of osseointegration of newly placed implants is currently unknown. It has been proposed that osseointegration would fail if an implant is luxated in bone more than 50 to 150 microns. Fibrous tissue, not bone, would form. This study investigated the quantity of various off-axial forces required to move a nonosseointegrated 4.3 x 13 mm implant 50 microns. The previously published pilot study for this study found that the amount of horizontal force required to displace an implant 50 microns was approximately 150 N. This study found that the force needed to move the implants 100 microns at a horizontal approach, 0 degrees, averaged 50 N, with a range of 23-79 N; at 22 degrees, averaged 52 N, with a range of 27-70 N; and at 60 degrees averaged 87 N, with a range of 33-105 N. PMID- 20017643 TI - Initial torque stability of a new bone condensing dental implant. A cohort study of 140 consecutively placed implants. AB - The aim of this paper was to determine the torque resistance of this new implant during placement in different types of bone, immediate placement into sockets, and in grafted bone. The torque at time of placement serves as an indication of initial stability, which is accepted as an important factor for implant osseointegration and immediate loading. Within a 13-month period, 140 NobelActive implants in 84 consecutive patients were placed into types I-IV bone in fresh sockets, and into grafted bone (both in maxillary sinuses and on the facial alveolar surfaces where bone had been lost). The final torque was measured with a manual torque control wrench as manufactured by Nobel Biocare for clinical use with this type of implant. One hundred forty implants with 3.5 to 5 mm diameters and 10 to 15 mm lengths were placed in different types of bone, either as delayed or immediate implants into fresh extraction sockets. These implants demonstrated a mean torque stability value of 50.8 Ncm. The average insertion torque for delayed implants was 49.7 Ncm. For immediate implants the average torque was 52.6 Ncm. Placement into soft bone was also favorable at an average of 47.9 Ncm. Typical straight walled and tapered implants generally exhibit 10 to 35 Ncm insertion torques. The NobelActive implant consistently reaches higher torque levels. This may indicate they are more favorably suited to early provisionalization and loading. Soft bone (type IV) did not seem to decrease significantly the torque of insertion of these implants. Further longer term studies are needed to investigate whether this indeed makes these implants more suited for early provisionalization and loading than traditional root form. Long term studies are also needed to investigate maintenance of bone levels surrounding these implants. PMID- 20017644 TI - Clinical evaluation of the NobelActive implant system: a case series of 107 consecutively placed implants and a review of the implant features. AB - The purpose of this paper is to (1) introduce the features of this new implant, (2) investigate the clinical benefits as advertised by the manufacturer in comparison with traditional root form implants, and (3) provide guidelines for its use. One hundred seven NobelActive implants were placed in 67 consecutive patients with type I-IV bone within 8 months. Cases also include implants placed in sinus grafts, ridges with insufficient thickness and facial bone loss and were placed with delayed and immediate loading. Parameters were assessed to determine whether we could confirm the manufacturer's statements on this implant system. Results obtained with 107 implants of 3.5, 4.3, and 5 mm diameters with 10- to 15 mm lengths placed in different types of bone with delayed and immediate loading demonstrated a final insertion torque from 15 to 70 Ncm. All types of bone allowed "redirection" of the implant but were limited in the bone with higher density. According to the manufacturer, this new design of the NobelActive implant has high initial stability, bone condensing properties, redirecting capability, built-in platform shifting, and dual-function prosthetic connections. After investigating these 5 statements within the limits of our study, we were able to confirm these claims, but with some recommendations for the clinical use and placement of these implants. PMID- 20017645 TI - Bone spreader technique: a preliminary 3-year study. AB - The purpose of this study was to observe the clinical outcome of bone spreading and standardized dilation of horizontally resorbed bone during immediate implant placement using a "screw-type" configuration of expansion and threadformers. Fifty-three patients were included in this study, and 41 edentulous areas in anterior and posterior maxillas were treated. Sixty-eight implants were placed using an insertion torque of at least 40 Ncm. Abutments were delivered 4 to 6 months after implant placement. The overall failure percentage was 4.41% (3 failures). A retrieved analysis of 1 implant removed at 3 years after placement demonstrated bone resorption down to the level of the third thread. The bone spreader technique is different from Summers' osteotome, both in clinical use and in armamentarium. The main advantage of the crest-expanding technique is that it is a less invasive procedure; the facial wall expands after the medullary bone is compressed against the cortical wall. Within the limits of this preliminary study, the cumulative survival rate for this method of implant placement is 95.58% at 3 years. This study confirms that a bone spreader used in the maxilla shows an unusually low failure rate after 3 years. PMID- 20017646 TI - Edentulous arch treatment with a CAD/CAM screw-retained framework and cemented crowns: a clinical case report. AB - This article describes the rehabilitation of a completely edentulous patient using a milled titanium implant framework and cemented crowns. This combined approach significantly offsets unsuitable implant position, alignment, or angulation, while ensuring the easy retrievability, repair, and maintenance of the prosthesis. Hence, the dual advantage of cemented-retained crowns reproducing appropriate esthetics and function, irrespective of where the screw access openings are located in the substructure, can be obtained, along with the splinting effect and management of soft and hard tissue deficits achievable with a screw-retained framework. PMID- 20017647 TI - A method for making the implant-supported record bases. AB - A method for fabrication of record bases which is used in implant dentistry has been described. This method is cost effective because it does not need extra equipment for making the maxillomandibular relations. This technique involves the application of plastic sprues that fit on the top of implants to hold the record bases and allows for easy recording of maxillomandibular relations with an implant-supported record base. PMID- 20017648 TI - Five-year study of implant placement in regenerated bone and rehabilitation with telescopic crown retained dentures: a case report. AB - Various implant-supported restorations have been used successfully for several decades to rehabilitate edentulous patients. Telescopic crowns are a common treatment modality used to connect dentures to natural teeth. Although previous findings indicate that telescopic crowns can be placed successfully on implants to support overdentures, only limited data are available on this treatment approach. Eight months after extraction of all nonsalvageable teeth and socket preservation, 11 implants were inserted into the mandible and maxillae of one patient. These implants were restored 4 months later using telescopic crown supported dentures. Bleeding on probing (BOP), plaque index (PI), clinical attachment (PAL), and radiographic bone level were evaluated over 5 years. All implants remained in function over the 5-year evaluation period. Radiography showed stable bone levels for all implants. No changes in BOP or PI (range, 2%-4% for both parameters) were observed over this time. The PAL deteriorated by 1.5 mm during the first 3 years, with no subsequent changes. We conclude that telescopic crowns can be used successfully as attachments for overdentures supported by implants in regenerated bone. PMID- 20017649 TI - How to review scientific manuscripts and clinical case reports for Journal of Oral Implantology. AB - Peer review is a vital component of the publication process for the Journal of Oral Implantology (JOI), as with any scientific or professional journal. The ability to write constructive critical reviews, therefore, is a valued skill of benefit to authors, editors, and the journal. The purpose of this article is to discuss techniques for crafting useful reviews. A review should address both broad issues of wording and perception. A review should include specific issues of what to look for in each section of research papers and clinical case reports. While the article is written with JOI in mind, most guidelines expressed therein are applicable to reviewing for any journal. PMID- 20017651 TI - Adiponectin in health and diseases: from metabolic syndrome to tissue regeneration. AB - IMPORTANCE OF THE FIELD: Adiponectin's importance as a regulator of metabolic functions has been assessed after observation of its downregulation in obese and diabetic patients. Decreased plasma adiponectin has been correlated with the onset of metabolic syndrome. Adiponectin has been proposed as an anti inflammatory factor involved in protection of microvascular endothelium from inflammation insults. AREAS COVERED IN THIS REVIEW: The pleiotropic role of adiponectin in health and disease, with the double aim of describing the complex network of molecules engaged in its signaling in different tissues and underlining the gaps in the available data which need further investigation. WHAT THE READER WILL GAIN: Recent studies suggest that adiponectin can act as a paracrine/autocrine factor during tissue regeneration as it is a stem cell factor in hematopoiesis and vasculogenesis, as well as in bone, liver and muscle regeneration. This review underlines the new findings on adiponectin in tissue regeneration, particularly skeletal muscle regeneration. TAKE HOME MESSAGE: The most recent findings suggest that adiponectin plays a role that goes beyond its well-known metabolic function. Indeed, this adipokine behaves as a stem factor for several tissues, giving a new perspective for its use as a tool for cell based therapy of diseased/injured tissues. PMID- 20017652 TI - Circulating fibronectin isoforms predict the degree of fibrosis in chronic hepatitis C. AB - OBJECTIVE: Hepatic stellate cells only produce fibronectin isoforms in disease states. The isoform-defining domains can be detected in the blood circulation. This study examines whether circulating levels of fibronectin isoforms show a relationship with liver fibrosis on histology in patients with chronic hepatitis C. MATERIAL AND METHODS: In a prospective study, 50 patients with chronic hepatitis C who underwent a liver biopsy were compared to 50 matched controls and 35 patients with other liver conditions. RESULTS: Circulating levels of the fibronectin isoforms were significantly higher in patients with chronic hepatitis C compared to healthy controls [oncofetal fibronectin (oFN) 2.45 +/- 0.17 versus 1.76 +/- 0.16 mg/l, P < 0.005; extra domain-A (EDA) 1.05 +/- 0.06 versus 0.86 +/- 0.06 mg/l, P < 0.05; and extra domain-B (EDB) 14.55 +/- 0.74 versus 9.31 +/- 0.58 mg/l, P < 0.001], even though total fibronectin was lower (198.9 +/- 3.5 versus 343.6 +/- 14.5 mg/l, P < 0.001). A correlation with the fibrosis score was found for both oFN (r = 0.46, P < 0.005) and EDA (r = 0.51, P < 0.001). The combination of an elevation in both markers (oFN and EDA) in the upper quartile was associated with a specificity of > 99% for predicting significant fibrosis (stages 2-4) and 95% for predicting advanced fibrosis (stages 3-4). A combination of decreased values in the lowest tertile for both markers had a specificity of 94% for excluding significant fibrosis. Based on these findings, 30% of the patients scheduled for a liver biopsy could be correctly classified as having or not having significant fibrosis. The remainder would have to proceed with a biopsy. CONCLUSION: Circulating fibronectin isoforms produced by activated stellate cells represent a viable marker for the presence of significant fibrosis or a lack thereof. PMID- 20017653 TI - Inflammatory alterations in excluded colon in rats: a comparison with chemically induced colitis. AB - Diversion colitis occurs commonly in the large bowel remnant after diversion of the fecal stream. Several experimental models of colitis have been described, but none examine the inflammatory alterations that can occur in experimentally defunctioned colons. This characterization could be useful in understanding pathophysiological aspects of diversion colitis, and in developing future therapeutic strategies. Thus, we evaluated the temporal inflammatory alterations in the defunctioned colon of rats by analyzing the histological results, infiltrating neutrophils, pro-inflammatory markers such as cyclooxygenase (COX)-2 and inducible nitric oxide synthase (iNOS), and DNA damage in isolated colonocytes. We compared the obtained data with those from hapten-induced colitis. The experimental diversion of the colon fecal stream induces diversion colitis characterized by an early inflammatory process with increased neutrophil infiltrate, and COX-2 and iNOS expression that resembles, in some aspects, the inflammatory characteristics of chemically induced colitis. After acute inflammation resolution, there was an increase in COX-2 and iNOS expression and the presence of lymphoid follicular hyperplasia and ulcerations, suggesting that diversion colitis can be experimentally established and useful for studying different pathophysiological aspects of this condition. PMID- 20017654 TI - Comment and reply on: The impact of body mass index and Heliobacter pylori infection on gastroesophageal reflux symptoms. PMID- 20017655 TI - Letter to the editor: Methisoprinol: a novel addition to hyper IgE arsenal. PMID- 20017657 TI - Pharmaceutical technology management--profitable business avenue. AB - Growing research expenditure, regulatory framework and generic erosion have forced pharmaceutical companies globally to resort to pharmaceutical technology management (PTM). Indeed, the pharmaceutical industry has witnessed the impact of innovative drug delivery and device technologies and their influence on business. PTM has given a new business insight with greater profits and enhancement of product franchise. Promising breakthrough technologies have not been able to reach a commercial platform largely owing to lack of capital at the preliminary stages of the product development program. Intellectual property plays a considerable role in protecting innovative technologies. Joint ventures and strategic alliances also become important for commercializing a new technology. The synergy of PTM with options of in-licensing is expected to infuse newer opportunities to the pharmaceutical business. PMID- 20017658 TI - Solid lipid nanoparticles for applications in gene therapy: a review of the state of the art. AB - IMPORTANCE OF THE FIELD: Gene therapy represents a new paradigm in the prevention and treatment of many inherited and acquired diseases, including genetic disorders, such as cystic fibrosis, haemophilia and many somatic diseases, such as tumours, neurodegenerative diseases and viral infections, such as AIDS. AREAS COVERED IN THIS REVIEW: Among a large array of non-viral transfection agents used for in-vitro applications, cationic SLNs are the topic of this review, being recently proposed as an alternative carrier for DNA delivery, due to many technological advantages such as large-scale production from substances generally recognized as safe, good storage stability and possibility of steam sterilization and lyophilisation. WHAT THE READER WILL GAIN: The authors give some information on the knowledge of intracellular trafficking and SLNs-DNA complex chemical physical properties reported until now in the literature. TAKE HOME MESSAGE: The future success of cationic SLNs for administration of genetic material will depend on their ability to efficiently cross the physiological barriers, selectively targeting a specific cell type in vivo and expressing therapeutic genes. PMID- 20017659 TI - Nanoparticle-based vaginal drug delivery systems for HIV prevention. AB - IMPORTANCE OF THE FIELD: Several strategies are being investigated for the prevention of heterosexual transmission of HIV. Of these, topical vaginal drug delivery systems, microbicides, are being actively pursued. HIV prevention by means of a topical microbicide has several drug delivery challenges. These challenges include the vaginal mucosal barriers and potential degradation of the drugs in the vaginal lumen due to pH and enzymes present. Also, new drugs being evaluated as microbicides have specific mechanisms of action, which in some cases require drug targeting to a specific site of action. Nanoparticles provide a delivery strategy for targeted or controlled delivery to the vagina which can be applied in the field of HIV prevention. AREAS COVERED IN THE REVIEW: This review summarizes nanoparticulate systems and their use in mucosal delivery to date. The sexual transmission of HIV along with the various targets to prevent transmission are discussed as well as the potential opportunities, challenges and advantages in using a nanoparticle-based approach for microbicidal drug delivery. WHAT THE READER WILL GAIN: This review provides a general understanding of vaginal drug delivery, its challenges, and nanoparticulate delivery systems. Additionally, insight will be gained as to the limited existing application of this technology to the field of HIV prevention. TAKE HOME MESSAGE: To date, few studies have been published that exploit nanoparticle-based microbicidal delivery to the vagina. The use of nanoparticles for vaginal drug delivery provides an approach to overcome the existing barriers to success. PMID- 20017660 TI - Overcoming the barriers in micellar drug delivery: loading efficiency, in vivo stability, and micelle-cell interaction. AB - IMPORTANCE OF THE FIELD: Spontaneously constructed from block copolymers in aqueous media, the polymer micelle has been extensively studied as a potential carrier of poorly water-soluble drugs, but cellular uptake pathways and stability of micelles in blood have not yet been clearly understood. An in-depth insight into the physical and biological behaviors of polymer micelles is necessitated for designing next-generation micelles. AREAS COVERED IN THIS REVIEW: This review suggests possible solutions to improve micellar drug loading capacity, scrutinizes the parameters influencing the micelle stability in blood, and also discusses the fate of micelles in cellular and in vivo environment, respectively. Direct and indirect evidences from the literatures mostly published after 90's were collected, analyzed and summarized. WHAT THE READER WILL GAIN: A critical analysis of micelle's stability in vivo and micelle-cell interaction is provided to highlight the key issues to be addressed to affirm that micelle can properly work as a drug carrier in clinical settings. TAKE HOME MESSAGE: With a clear understanding of its behaviors in biological environment, the polymer micelle is a promising nanocarrier for chemotherapy. PMID- 20017661 TI - Chemical permeation enhancers for transbuccal drug delivery. AB - IMPORTANCE OF THE FIELD: The buccal drug delivery system has been accepted as a potential non-invasive route of drug administration, with the advantages of avoidance of the first-pass metabolism, sustained therapeutic action and better patient compliance. However, transmucosal delivery of drugs by means of the buccal route is still very challenging. The main obstacles derive from the limited absorption area and from the barrier properties of the mucosa that have to be overcome for successful delivery drug molecules to the systemic circulation by this route. AREAS COVERED IN THIS REVIEW: One long-standing approach for improving buccal drug delivery uses buccal absorption promoters, also called permeation enhancers. This requisite has fostered the study of permeation enhancers that will safely alter the permeability restrictions of the buccal mucosa. This review includes various classes of transmucosal chemical permeation enhancers and their mechanism of action. As enhancers influence drug delivery, further exploration of these compounds is required to understand their modifying action on the properties of buccal mucosa. WHAT THE READER WILL GAIN: This review will help the readers in the selection of a suitable enhancer(s) for improving the buccal drug delivery for future endeavor. TAKE HOME MESSAGE: The authors imagine new buccal formulations bearing permeation enhancer(s) being commercialized in the coming years. PMID- 20017662 TI - Various drug delivery approaches to the central nervous system. AB - IMPORTANCE OF THE FIELD: The presence of the blood-brain barrier (BBB), an insurmountable obstacle, in particular, and other barriers in brain and periphery contribute to hindrance of the successful diagnosis and treatment of a myriad of central nervous system pathologies. This review discusses several strategies adopted to define a rational drug delivery approach to the CNS along with a short description of the strategies implemented by the authors' group to enhance the analgesic activity, a CNS property, of chimeric peptide of Met-enkephalin and FMRFa (YGGFMKKKFMRFa-YFa). AREAS COVERED IN THIS REVIEW: Various approaches for drug delivery to the CNS with their beneficial and non-beneficial aspects, supported by an extensive literature survey published recently, up to August 2009. WHAT THE READER WILL GAIN: The reader will have the privilege of gaining an understanding of previous as well as recent approaches to breaching the CNS barriers. TAKE HOME MESSAGE: Among the various strategies discussed, the potential for efficacious CNS drug targeting in future lies either with the non invasively administered multifunctional nanosystems or these nanosystems without characterstics such as long systemic circulating capability and avoiding reticuloendothelial system scavenging system of the body, endogenous transporters and efflux inhibitors administered by convection-enhanced delivery. PMID- 20017663 TI - In reply to letter to the editor by Dr Willems et al. re: Eccles et al. Change in diffusion weighted MRI during liver cancer radiotherapy: Preliminary observations. PMID- 20017664 TI - FDG-PET-CT for staging of high-risk breast cancer patients reduces the number of further examinations: A pilot study. AB - AIM: To determine the additional value of FDG-PET-CT as compared to conventional staging (CS) in high-risk breast cancer patients. PATIENTS AND METHODS: Thirty one high-risk breast cancer patients, 14 of whom had recurrent breast cancer, were included in this study, which took place between June 2005 and March 2008. None of the patients had clinical signs of distant metastases. FDG-PET-CT scanning was added to CS, which consisted of a chest x-ray, liver ultrasonography or CT, and bone scintigraphy. Median follow-up was 17 months (6-41 months). FDG PET-CT was considered to have additional value to CS if it led to a change in treatment plan or if it made additional examinations to confirm or deny findings on CS unnecessary. RESULTS: FDG-PET-CT was considered to have additional value to CS in 13 patients (42% [95% CI: 23-61]). In five patients (16% [95% CI: 1-31]), FDG-PET-CT led to a change in treatment plan by identifying nodal metastases in the internal mammary chain (IMC; N = 3) or in the mediastinum (N = 2). In nine patients (29% [95% CI: 11-47]), FDG-PET-CT would have prevented the need for additional examinations; in seven of these nine patients, distant metastases were suggested in bone or liver on CS, but these did not show FDG uptake. CONCLUSIONS: FDG-PET-CT was found to have additional value to CS in 42% of the patients. To optimize cost-effectiveness, the main challenge now is to improve the selection of patients in whom FDG-PET-CT has additional value to CS. PMID- 20017665 TI - Pharmacogenomics and the prevention of mother-to-child transmission of HIV. PMID- 20017667 TI - Research highlights. Genetics in renal transplantation: a recent tale of chemokines, enzymes, drugs and HLA. PMID- 20017668 TI - Institutional profile. UNC Institute for Pharmacogenomics and Individualized Therapy: interdisciplinary research for individual care. AB - The Institute for Pharmacogenomics and Individualized Therapy (IPIT) at the University of North Carolina at Chapel Hill (NC, USA) is a collaborative, multidisciplinary unit that brings together faculty from different disciplines and crosses the traditional departmental/school structure to perform pharmacogenomics research. IPIT investigators work together towards the goal of developing therapies to enable the delivery of individualized medical care. The NIH-supported Comprehensive Research on Expressed Alleles in Therapeutic Evaluation (CREATE) group leads the field in the evaluation of pathways regulating drug activity, and also provides a foundation for future IPIT research. IPIT members perform bench research, clinical cohort analysis and prospective clinical intervention studies, research on the integration of pharmacogenomic therapy into practice and research to foster global health pharmacogenomics application through the Pharmacogenetics for Every Nation Initiative. IPIT Investigators are actively incorporating a pharmacogenomics curriculum into existing teaching programs at all levels. PMID- 20017669 TI - Nevirapine-induced hepatotoxicity and pharmacogenetics: a retrospective study in a population from Mozambique. AB - AIMS: Nevirapine is widely used to treat HIV-1 infection to prevent mother-to child transmission; unfortunately adverse drug reactions have been reported. Our aim was to identify genes/variants involved in nevirapine-induced hepatotoxicity. MATERIALS & METHODS: Patients from Mozambique, 78 with nevirapine-induced hepatotoxicity and 78 without adverse events, were genotyped for ABCB1, CYP2B6, CYP3A4 and CYP3A5 gene variants. We conducted a case-control association study and a genotype/phenotype correlation analysis. RESULTS: The ABCB1 c.3435C>T SNP was associated with hepatotoxicity (p = 0.038), with the variant T allele showing a protective effect (odds ratio: 0.42). Moreover, four SNPs in the CYP2B6 and CYP3A5 genes resulted significantly correlated with transaminase values. In particular, for the CYP2B6 c.983T>C SNP, the difference in the alanine aminotransferase mean values were highly significant between TT and TC genotypes (p < 0.001). CONCLUSION: Our preliminary results confirm the contribution of the ABCB1 c.3435C>T SNP in nevirapine-induced hepatotoxicity risk and, at the same time, suggest the necessity for further studies. PMID- 20017670 TI - Genetic polymorphisms in GST genes and survival of colorectal cancer patients treated with chemotherapy. AB - Glutathione S-transferases (GSTs) participate in the detoxification of chemotherapeutic agents. Genetic polymorphisms in GST genes (GSTP1 Ile105Val, copy-number variants of GSTM1 and GSTT1) that lead to diminished enzyme activity have been associated with increased chemotherapeutic treatment benefit in colorectal cancer patients. AIMS: We assessed the effect of genetic polymorphisms in GST genes on survival in colorectal cancer patients treated with adjuvant/palliative chemotherapy. As GSTs participate in the metabolism of platinum metabolites, we also assessed the association between genetic variants in GST genes and survival of colorectal cancer patients who received treatment with oxaliplatin. MATERIALS & METHODS: We followed 338 colorectal cancer patients treated with chemotherapy for a median of 36.4 months since treatment start. A total of 65 of the patients received treatment with oxaliplatin. Polymorphisms were genotyped by fluorescence-based melting curve analysis (GSTP1 Ile105Val), a relative quantification method (copy-number variants of GSTM1 and GSTT1), and PCR followed by gel electrophoresis (null/non-null genotypes for GSTM1 and GSTT1). Associations between genotypes and overall survival were assessed using Kaplan Meier curves and Cox proportional hazards regression. RESULTS: As hypothesized, GSTM1 copy number variant was inversely associated with survival in colorectal cancer patients treated with chemotherapy. Mortality was significantly reduced in patients with one GSTM1 copy (hazard ratio: 0.45, 95% CI: 0.23-0.90, p = 0.02) and nonsignificantly reduced in those with the null genotype (HR: 0.67, 95% CI: 0.35-1.27, p = 0.22) compared with carriers of two copies. Both GSTP1 genotype and GSTT1 genotype were not associated with survival. CONCLUSION: This is the first study to provide suggestive evidence for an effect of copy-number variation of GSTM1 on survival in colorectal cancer patients who received chemotherapy. Large studies are warranted to establish the impact of GST genotypes on treatment outcome in colorectal cancer patients. PMID- 20017671 TI - CYP2D7-2D6 hybrid tandems: identification of novel CYP2D6 duplication arrangements and implications for phenotype prediction. AB - AIMS: Allelic variants of cytochrome P450 CYP2D6 (CYP2D6), such as gene deletion, duplication, multiplication and conversion, contribute to the wide range of CYP2D6 activity. Novel gene arrangements were discovered and characterized. MATERIALS & METHODS: DNA from 32 Caucasian and 59 African-American duplication positive subjects were analyzed by long-range PCR and genotyping to detect CYP2D7 2D6 hybrid tandem alleles. Novel allelic variants were sequenced and a strategy for the detection and analysis of hybrid genes was refined. RESULTS: CYP2D7-2D6 hybrid tandem alleles were identified in one African-American and four Caucasian subjects. Three novel hybrid genes were found on CYP2D6*1 and CYP2D6*2 duplication backgrounds and designated CYP2D6*76, *77 and *78. CYP2D7 to 2D6 conversion occurred in introns 1 and 4, and exon 9. All carried a T-insertion in exon 1 abolishing activity. In Caucasians, four out of 33 (12%) of the duplication-positive alleles were hybrid tandems, three CYP2D6*77 + *2 and one CYP2D6*78 + *2. By contrast, in African-Americans only one of 60 duplication positive alleles was identified as a hybrid tandem. This allele was designated CYP2D6*76 + *1. CONCLUSION: Hybrid tandem alleles occur infrequently (<0.25%) in Caucasians, but may explain why not every subject with a CYP2D6 duplication presents with an ultrarapid metabolizer phenotype. PMID- 20017672 TI - TaqMan low-density arrays and analysis by artificial neuronal networks predict response to neoadjuvant chemoradiation in esophageal cancer. AB - AIMS: Neoadjuvant radiochemotherapy of locally advanced esophageal cancer is only effective for patients with major histopathological response. A total of 17 genes were selected to predict histopathologic tumor response to chemoradiation (cisplatin, 5-fluorouracil, 36 Gy). MATERIALS & METHODS: For gene-expression analysis quantitative TaqMan low-density arrays were applied. Expression levels in pretreatment biopsies of 41 patients (cT2-4, Nx, M0) were compared with the degree of histopathologic regression in resected specimens applying univariate, multivariate and artificial neuronal network analyses. RESULTS: Dihydropyrimidine dehydrogenase was identified as an independent predictor associated with major response (p < 0.002). Multivariate analysis of the marker combination provided response prediction with 75.0% sensitivity, 81.0% specificity and 78.1% accuracy. Artificial neuronal network analysis was the best predictive model for major histopathologic response (80% sensitivity, 90.5% specificity and 85.4% accuracy), representing a clinically practical system. CONCLUSION: Low-density-array RT-PCR analyzed by artificial neuronal network predicts histopathologic response to neoadjuvant chemoradiation in our patient collective, and could be used to further individualize treatment strategies in esophageal cancer. PMID- 20017673 TI - Pharmacogenetics in reproductive and perinatal medicine. AB - The clinical application of pharmacogenetics has been well accepted by some medical specialties, but not all. The aim of this review is to discuss the current use of pharmacogenetics in reproductive and perinatal medicine and to highlight areas where pharmacogenetics may be able to help in the future to predict response to medicines in terms of efficacy and safety. This applies to drugs that are specific to pregnancy and reproduction, as well as drugs prescribed for the treatment of medical disorders in pregnancy. Our review points out the need for well-designed clinical studies on the efficacy and safety of medicines used in women of childbearing age in order to define the additional utility provided by pharmacogenetic testing. PMID- 20017674 TI - Meaningful use of pharmacogenomics in health records: semantics should be made explicit. AB - The recent emphasis on 'meaningful use' of electronic health records in health information technology reforms (e.g., as in the US stimulus package) can leverage the pharmacogenomics field. In order for clinical trials outcomes, based on pharmacogenomics research, to be meaningfully and effectively used in clinical practice there is a need to make health semantics explicit. Often, semantics is merely implicit in both the research and practice worlds and is buried in unstructured and disconnected descriptions of the data or just in the heads of human experts. Meaningful semantics includes rich metadata, but more importantly, the context of each discrete data item and how it relates to other data items in a specific dataset, as well as how it fits within the entire health record of an individual and how it references up-to-date clinical knowledge. Properly-built electronic health records systems based on standards could provide meaningful semantics on the healthcare side, while the fields of research and clinical trials need to come closer to healthcare in its data and knowledge representations in a way that lends itself to personalized medicine. The purpose of this review is to explore how evidence created by pharmacogenomics can be meaningfully delivered to healthcare through new approaches, such as electronic health records systems and information models. PMID- 20017675 TI - Clinical pharmacology and pharmacogenetics in a genomics era: the DMET platform. AB - While no genome-wide pharmacogenetics study has yet been published, the field of pharmacogenetics is moving towards exploratory, large-scale analyses of the interaction between genetic variation and drug treatment. The Drug Metabolizing Enzymes and Transporters (DMET) platform offers a standardized set of 1936 variants in 225 genes related to drug absorption, distribution, metabolism and elimination that is useful to scan the genome for previously unknown associations between variation in absorption, distribution, metabolism and elimination genes and pharmacokinetic and pharmacodynamic outcomes of drug treatment. The purpose of this review is to put the DMET platform into context within the current study designs that have been used in pharmacogenetics, and to explore the role that DMET has played - and will play - in future pharmacogenetics studies. PMID- 20017676 TI - Pathology practice and pharmacogenomics. AB - New technologies emerging from the Human Genome Project and the rapidly expanding direct-to-consumer DNA testing have provided a challenging environment for the entry of pharmacogenomics into clinical practice. The traditional pathology laboratory, which is centered around a referring clinician and the patient, is also being reshaped by these developments. These changes are occurring as the shrinking health dollar imposes a greater focus on preventative medicine. For the latter to benefit from genomics requires the community to be engaged, and the development of the disease process identified earlier than otherwise possible. These two prerequisites are now being sought through the concept of 'personalized medicine'. Much has been occurring in a relatively short time frame, and as a consequence, education becomes a major rate-limiting step to change. PMID- 20017679 TI - What is the influence of background noise and exercise on the listening levels of iPod users? AB - The widespread use of portable listening devices (PLDs) has increased concern about the potential for hearing impairment caused by their use. The current study investigated the effects of external noise and exercise on the use of PLDs. The 24 participants listened to the same song on an iPod during rest-in-quiet, rest in-noise, and exercise-in-noise conditions. Preferred listening levels (PLLs) were recorded and participants' maximum noise doses were calculated. Participants selected significantly higher listening levels in both noise conditions than in the quiet condition. The variability of volume selection was reduced significantly in the noise conditions. The maximum daily noise dose would have been exceeded by seven participants in the rest-in-noise condition and by eight in the exercise-in-noise condition compared to one participant in the rest-in quiet condition. These results indicated that increased background noise causes individuals to increase the volume on their PLDs to potentially dangerous levels and that increased noise alone was not the only factor affecting the participants as the addition of exercise induced even further increases in PLLs. PMID- 20017677 TI - ARMS test for diagnosis of CYP2C9 and VKORC1 mutation in patients with pulmonary embolism in Han Chinese. AB - AIMS: VKORC1 and CYP2C9 are important genetic factors affecting warfarin dose requirement. Our aim is to establish a simple, rapid and economical method to detect SNPs in the two genes. MATERIALS & METHODS: Primer Premier 5 was used and a normal primer, a mutational primer and a common primer have been designed using the amplification refractory mutation system for VKORC1 c.-1639G>A (rs9923231), CYP2C9*3 c.1075A>C (rs1057910) and CYP2C9*13 c.269T>C substitutions. The amplification refractory mutation system (ARMS) assay was validated by the restriction enzyme cleavage method of reference or direct sequencing. RESULTS: The ARMS primers designed can distinguish between heterozygotes and homozygotes successfully. In the Han Chinese population, patients with pulmonary embolism allele frequencies of CYP2C9*3 c.1075A>C (rs1057910), *13 c.269T>C and VKORC1 1639G>A (rs9923231) are 4.3, 0.7 and 8.6%, respectively. CONCLUSION: The ARMS-PCR method is a simple, economical method that can be used for the rapid detection of SNPs in VKORC1 and CYP2C9. PMID- 20017680 TI - The detection of adult cortical auditory evoked potentials (CAEPs) using an automated statistic and visual detection. AB - The detection of adult cortical auditory evoked potentials (CAEPs) can be challenging when the stimulus is just audible. The effectiveness of a statistic compared with expert examiners in (1) detecting the presence of CAEPs when stimuli were present, and (2) reporting the absence of CAEPs when no stimuli were present, was investigated. CAEPs recorded from ten adults, using two speech-based stimuli, five stimulus presentation levels, and non-stimulus conditions, were given to four experienced examiners who were asked to determine if responses to auditory stimulation could be observed, and their degree of certainty in making their decision. These recordings were also converted to multiple dependent variables and Hotelling's T2 was applied to calculate the probability that the mean value of any linear combination of these variables was significantly different from zero. Results showed that Hotelling's T2 was equally sensitive to the best of individual experienced examiners in differentiating a CAEP from random noise. It is reasonable to assume that the difference in response detection for a novice examiner and Hotelling's T2 would be even greater. PMID- 20017681 TI - Do children with reading delay benefit from the use of personal FM systems in the classroom? AB - FM systems have been used to compensate for poor signal-to-noise ratios in classrooms. This study evaluates benefits of a 6-week trial of personal FM systems used during the school day for children with reading delay aged 6-11 years, using a randomized control design. Teachers and children completed the LIFE-UK questionnaire. Test-retest reliability of the LIFE-UK children's version was confirmed in a separate group of 18 children from the same school. The 23 children in the FM group had significantly improved teacher ratings, and the children's ratings of classroom listening for difficult situations were significantly better after the trial. These changes did not occur for the 23 control-group children. Most children (92%) commented positively about the FM after the trial. It is likely that a longer FM trial or a specific reading intervention combined with FM will be required for the benefits of enhanced listening to affect performance on standardized reading tests. PMID- 20017682 TI - Environmental noise reduction configuration: Effects on preferences, satisfaction, and speech understanding. AB - The effects of four configurations of an environmental noise reduction (ENR) algorithm on preferences, speech understanding, and satisfaction were investigated. The gain reduction at 0 dB modulation depth was either 10 dB in all channels (ENR StrongFlat) or shaped from 2-10 dB across channels according to a speech importance function (ENR MildSII). This gain reduction was either invariant (ENR Constant) or varied with (ENR Variable) the noise level. Ten hearing-impaired participants blindly compared pairs of configurations in real world situations and recorded their preferences. Sentence reception thresholds (SRTs) were measured in quiet and noise, and satisfaction was rated with speech in noise. Half of the participants preferred ENR MildSII and half preferred ENR StrongFlat. All preferred ENR Variable to ENR Constant. Overall, the preferred ENR configuration was preferred to ENR off in 90% of responses. No statistically significant effect on SRTs was found, but a clinically significant effect of up to 2 dB could not be ruled out from the available data. ENR significantly improved satisfaction for listening comfort, ease of speech understanding, and sound quality. PMID- 20017683 TI - A high rate n-of-m speech processing strategy for the first generation Clarion cochlear implant. AB - An n-of-m speech coding strategy has been developed for the Clarion Cochlear Implant Series 1.x (1.0 & 1.2). The basic principle is to reduce the number of stimuli per cycle, by neglecting the less significant spectral components, and to concentrate on the more dominant frequency bands. In this study 20 subjects, implanted with a Clarion device, used an n-of-m strategy at 1666 pps per channel. The outcomes using this strategy were compared with the outcomes using conventional processing (CIS at 833 pps/channel). Eight of the 20 subjects underwent additional testing with the n-of-m strategy with the rate set at 833 pps/channel. Using the n-of-m strategy at 1666 pps showed statistically significant improvement in performance over the CIS strategy, with 16 of the 20 subjects achieving better results. However, there was no statistically meaningful difference in performance between n-of-m at 833 pps and the CIS strategy running at the same rate. Results therefore suggest that n-of-m strategy can be an alternative to CIS, particularly when the implant hardware limits the overall stimulation rate. PMID- 20017684 TI - A biomechanical ear model to evaluate middle-ear reconstruction. AB - In order to evaluate the efficiency of middle-ear prostheses in near-real conditions, an artificial model was developed that approximately simulates the actual geometrical and biomechanical properties of the ear system (excluding the ossicular chain). The sound transmission characteristics of selected commercial middle-ear prostheses and of a synthetic test material were measured using laser Doppler vibrometry (LDV) in this model. The model's realistic properties enabled clinical tympanometry to be used to control the stiffness. In addition the influences of the implant mass on transmission characteristics were investigated. With an averaged displacement between 10 and 100 nm/Pa up to 2000 Hz, the transmission characteristic of the model was comparable with data obtained from the intact middle ear in temporal bone experiments. From the acoustical point of view, no significant material-specific differences could be found. Increasing the mass of the implants to more than 50 mg results in poorer acoustic transmission. In general, changes to the stiffness involving compliance values greater than 3.5 ml and smaller than 0.2 ml led to poorer acoustic transmission. PMID- 20017686 TI - Stem cell nations working together for a stem cell world. PMID- 20017687 TI - Stem cell charter. PMID- 20017690 TI - The common path: tumor suppression in the generation of iPS cells and cancer stem cells. PMID- 20017691 TI - McGowan Institute for Regenerative Medicine. PMID- 20017692 TI - Tracking the rise of stem cell tourism. AB - AIMS: Driven by hype surrounding stem cell research, a number of clinics around the world currently offer 'stem cell therapies' to patients. These unproven interventions have attracted policy interest owing to the risks they may pose to patients and to the progress of legitimate translational stem cell research, yet remarkably little data exists about the patients who undergo these unproven therapies or their experiences. We sought to characterize this patient population. MATERIALS & METHODS: We developed a comprehensive data set of blogs written by patients (or their caretakers) about their experiences with unproven stem cell therapies. RESULTS & CONCLUSIONS: Analyzing these data suggests that unproven stem cell therapies are increasing rapidly in popularity and are attracting a wide range of patients--both young and old and with a diverse collection of medical conditions. These results should help clinicians advise individual patients and help policymakers devise strategies to mitigate the risks these treatments pose. PMID- 20017693 TI - Human umbilical cord blood mononuclear cells decrease fibrosis and increase cardiac function in cardiomyopathy. AB - AIMS: We investigated whether human umbilical cord blood mononuclear cells (HUCBC) can limit progressive cardiomyopathy in TO2 hamsters. MATERIALS & METHODS: A total of 22 TO2 1-month-old hamsters were treated with intramyocardial HUCBC, 4 x 10(6) in Isolyte, and 23 TO2 1-month-old hamsters were treated with intramyocardial Isolyte. A total of 16 1-month-old F1B hamsters served as controls and received intramyocardial Isolyte. Echocardiograms were performed on all hamsters prior to and monthly after treatment for 6 months. Heart tissues were then stained with hematoxylin and eosin, Masson's Trichrome and human leukocyte antibody. RESULTS: In F1B hamsters, left ventricular fractional shortening (FS) and ejection fractions (EF) did not significantly decrease over 6 months. By contrast, in Isolyte-treated TO2 hamsters, FS decreased from 56.2 +/- 1.0% to 19.7 +/- 3.2% and EF decreased from 89.5 +/- 1.4% to 44.9 +/- 5.9% at 6 months (both p < 0.0001). The FS and EF in HUCBC-treated TO2 hamsters also progressively decreased over 6 months but the changes were more gradual, especially during the first month after HUCBC treatment when FS was 52.0 +/- 1.5% and EF was 89.5 +/- 1.4%, which was not significantly different from the FS and EF in the F1B hamsters. Moreover, in the HUCBC-treated hamsters, the FS and EF were 20-30% greater than FS and EF in Isolyte TO2 hamsters at 3 and 5 months (p < 0.01). In Isolyte-treated TO2 hamsters at 6-7 months, fibrosis involved 30.0 +/- 5.0% of left ventricle and 35.0 +/- 5.0% of septum. By contrast, in HUCBC-treated hamsters, fibrosis involved only 6.5 +/- 2.3% of the left ventricle and 6.3 +/- 1.8% of septum (p < 0.05). The average number of blood vessels per myocardial microscopic field in HUCBC-treated hearts was 53.5 +/- 0.8 versus 46.2 +/- 3.0 in Isolyte-treated TO2 hearts (p < 0.05). CONCLUSION: HUCBC, when given as a single intramyocardial injection, can limit fibrosis and increase heart function over the short term in TO2 hamsters with cardiomyopathy. PMID- 20017694 TI - Development of a calcium-chelating hydrogel for treatment of superficial burns and scalds. AB - AIMS: Superficial burns and scalds are usually managed conservatively with traditional dressings. Failure to heal within 3 weeks leads to their management by skin grafting. Our aim was to develop a biomaterial to actively promote keratinocyte migration in superficial burns by modulating local cation concentrations to accelerate keratinocyte migration and deter wounds from contracting, thus potentially reducing the number of such wounds requiring grafting. MATERIALS & METHODS: We investigated polymeric hydrogels for their Ca(2+) chelating properties and enhancement of keratinocyte migration in human tissue-engineered skin models. RESULTS: Dimethylaminoethyl methacrylate:methacrylic acid hydrogel coupled with elevated [Mg(2+)] reduced media [Ca(2+)], potentiating keratinocyte migration in tissue-engineered skin models, it also significantly reduced wound model contraction. CONCLUSION: Dimethylaminoethyl methacrylate:methacrylic acid hydrogels could promote wound healing and reduce wound contraction, a significant complication in burn wound healing. PMID- 20017695 TI - Progenitor cell therapy for traumatic brain injury: effect of serum osmolarity on cell viability and cytokine production. AB - INTRODUCTION: The potential translation of mesenchymal stem cell (MSC) therapy into a multimodal protocol for traumatic brain injury requires evaluation of viability and cytokine production in a hyperosmolar environment. Optimization of MSC therapy requires delivery to the target area without significant loss of cellular function or viability. No model evaluating the potential efficacy of MSC therapy at varying osmolarities currently exists. METHODS: Rat MSCs were characterized with flow cytometric immunophenotyping. MSCs (passage 3) were placed in culture with multipotent adult progenitor cell media at varying osmolarities (250, 270, 290, 310, 330, 350 and 370 mOsm) potentially found with hypertonic saline infusion. After culture for 24 h, cellular viability was measured using flow cytometry (n = 6). Next, brain tissue supernatant was harvested from both normal rat brains and injured brains 6 h after cortical injury. Subsequently, MSCs were placed in culture with multipotent adult progenitor cell media +/- 20% normal brain or injured brain supernatant (at the aforementioned osmolarities) and allowed to remain in culture for 24 h (n = 11). At this point, media supernatant cytokine levels were measured using a multiplex cytokine assay system. RESULTS: MSCs showed no clinically significant difference in viability at 24 h. MSCs cultured with 20% injured brain supernatant showed an decrease in proinflammatory cytokine production (IL-1alpha and IL-1beta) with increasing osmolarity. No difference in anti-inflammatory cytokine production (IL 4 and IL-10) was observed. CONCLUSION: Progenitor cell therapy for traumatic brain injury may require survival and activity in a hyperosmolar environment. Culture of MSCs in such conditions shows no clinically significant effect on cell viability. In addition, MSC efficacy could potentially be enhanced via a decrease in proinflammatory cytokine production. Overall, a multimodal traumatic brain injury treatment protocol based upon MSC infusion and hypertonic saline therapy would not negatively affect progenitor cell efficacy and could be considered for multicenter clinical trials. PMID- 20017696 TI - In vivo neural stem cell imaging: current modalities and future directions. AB - Neural stem cells have been proposed as a promising therapy for treating a wide variety of neuropathologies. While several studies have demonstrated the therapeutic benefits of neural stem cells, the exact mechanism remains elusive. In order to facilitate research efforts to understand these mechanisms, and before neural stem cell-based therapies can be utilized in a clinical context, we must develop means of monitoring these cells in vivo. However, because of tissue depth and the blood-brain barrier, in vivo imaging of neural stem cells in the brain has unique challenges that do not apply to stem cells for other purposes. In this paper, we review contemporary methods for in vivo neural stem cell imaging, including MRI, PET and optical imaging techniques. PMID- 20017697 TI - Characterizing functional stem cell-cardiomyocyte interactions. AB - Despite the progress in traditional pharmacological and organ transplantation therapies, heart failure still afflicts 5.3 million Americans. Since June 2000, stem cell-based approaches for the prevention and treatment of heart failure have been pursued in clinics with great excitement; however, the exact mechanisms of how transplanted cells improve heart function remain elusive. One of the main difficulties in answering these questions is the limited ability to directly access and study interactions between implanted cells and host cardiomyocytes in situ. With the growing number of candidate cell types for potential clinical use, it is becoming increasingly more important to establish standardized, well controlled in vitro and in situ assays to compare the efficacy and safety of different stem cells in cardiac repair. This article describes recent innovative methodologies to characterize direct functional interactions between stem cells and cardiomyocytes, aimed to facilitate the rational design of future cell-based therapies for heart disease. PMID- 20017703 TI - Neurokinin B and urotensin II levels in pre-eclampsia. AB - OBJECTIVE: To verify neurokinin B (NKB) and urotensin II (UII) plasma levels in pre-eclampsia (PE) and to determine the relationship between plasma NKB and UII levels. METHOD: A total of 60 women in the third trimester of pregnancy were recruited, 40 women with PE (study patients) and 20 age- and BMI-matched normotensive women (healthy controls). They were divided into three groups: the 20 normotensive pregnant women (Group 1); 20 women with mild PE (Group 2); 20 women with severe PE (Group 3). The plasma levels of NKB and UII were measured simultaneously by enzyme-linked immunosorbent assay. RESULTS: Compared with controls, levels of NKB were significantly higher in women with mild or severe PE (p < 0.01 for both groups), levels of UII were significantly higher in women with mild or severe PE (p < 0.01 for both groups). Moreover, there was a positive correlation between plasma levels of NKB and UII in pre-eclamptic women (r = 0.783, p < 0.01). CONCLUSIONS: These findings suggest that there was an elevation of NKB and UII in the plasma of pre-eclamptic women. NKB and UII may be involved in the pathophysiology of PE and associated with the development of severe disease. PMID- 20017698 TI - Biomaterials for vascular tissue engineering. AB - Cardiovascular disease is the leading cause of mortality in the USA. The limited availability of healthy autologous vessels for bypass grafting procedures has led to the fabrication of prosthetic vascular conduits. While synthetic polymers have been extensively studied as substitutes in vascular engineering, they fall short of meeting the biological challenges at the blood-material interface. Various tissue engineering strategies have emerged to address these flaws and increase long-term patency of vascular grafts. Vascular cell seeding of scaffolds and the design of bioactive polymers for in situ arterial regeneration have yielded promising results. This article describes the advances made in biomaterials design to generate suitable materials that not only match the mechanical properties of native vasculature, but also promote cell growth, facilitate extracellular matrix production and inhibit thrombogenicity. PMID- 20017704 TI - Uterine artery notch is associated with increased placental endothelial nitric oxide synthase and heat shock protein. AB - OBJECTIVE: This study was undertaken to evaluate whether uterine artery notch is associated with placental expression of endothelial nitric oxide synthase (eNOS), Heat shock protein 70 (Hsp70), and Lectin-like oxidized low-density lipoprotein receptor (Lox) in pregnant women. STUDY DESIGN: Uterine artery Doppler was performed in 30 pregnant women divided into two groups with or without the presence of uterine artery notches. After delivery, placental expression of eNOS, Hsp70, and Lox were evaluated with western blot analysis. RESULTS: Higher levels of placental eNOS (p = 0.02) and Hsp70 (p = 0.001) were expressed in the group with uterine artery notch compared with the group without uterine artery notch. There was no significant difference for placental Lox expression between group with uterine artery notch and without uterine artery notch (p = 0.17). CONCLUSION: Our results suggest that uterine artery notch is associated with increased placental expression of eNOS and Hsp in pregnant women. PMID- 20017705 TI - Comment and reply on: prediction of the date of delivery based on first trimester ultrasound measurements: an independent method from estimated date of conception. PMID- 20017706 TI - The effects of alpha-zearalanol and estradiol benzoate on expression of c-myc, c fos and epidermal growth factor receptor mRNAs in breast tissues implanted into nude mice. AB - This study was designed to evaluate the effects of a novel phytoestrogen, alpha zearalanol (alpha-ZAL), and estradiol benzoate (B-E2), on c-myc, c-fos, and EGFR expression in normal human breast tissues implanted into nude mice. A xenograft model, pieces of normal human breast tissue implanted subcutaneously into 9-10 week-old athymic nude mice, was established. The mice were divided into five groups subjected to the following treatments: normal saline (Controls); alpha-ZAL at 1 and 5 mg/kg; and estradiol benzoate (B-E2) at 1 and 5 mg/kg. Treatment was given every other day, and human breast tissues were removed for experiments after treatment for 30 days. The expression of c-myc, c-fos, and EGFR mRNAs were determined by in situ hybridization. alpha-ZAL decreased expression of c-myc (p < 0.05). About 1 mg/kg alpha-ZAL increased EGFR expression (p < 0.05) and two dosage of alpha-ZAL increased c-fos expression (p < 0.01) compared with control. B-E2 significantly increased expression of c-myc, c-fos, and EGFR mRNAs expression compared with controls (p < 0.01). The extents of the increases in EGFRmRNA expression induced by alpha-ZAL and in c-fos mRNA by 5 mg/kg alpha-ZAL were lower than those induced by B-E2 (p < 0.01). These data suggest that the phytoestrogen alpha-ZAL may be safer than estrogen on breast. PMID- 20017707 TI - Tibolone revisited: still a good treatment option for healthy, early postmenopausal women. PMID- 20017699 TI - Stem cell paracrine actions and tissue regeneration. AB - Stem cells have emerged as a key element of regenerative medicine therapies due to their inherent ability to differentiate into a variety of cell phenotypes, thereby providing numerous potential cell therapies to treat an array of degenerative diseases and traumatic injuries. A recent paradigm shift has emerged suggesting that the beneficial effects of stem cells may not be restricted to cell restoration alone, but also due to their transient paracrine actions. Stem cells can secrete potent combinations of trophic factors that modulate the molecular composition of the environment to evoke responses from resident cells. Based on this new insight, current research directions include efforts to elucidate, augment and harness stem cell paracrine mechanisms for tissue regeneration. This article discusses the existing studies on stem/progenitor cell trophic factor production, implications for tissue regeneration and cancer therapies, and development of novel strategies to use stem cell paracrine delivery for regenerative medicine. PMID- 20017708 TI - Association of interleukin-18 gene polymorphisms with calcium oxalate kidney stone disease. AB - OBJECTIVE: The interleukin-18 (IL-18) encoding gene has three common single nucleotide polymorphisms at -607C/A, -137G/C and +105A/C, which have been reported to be associated with several diseases. The aim of this study is to test whether IL-18 polymorphisms could act as genetic markers for renal stone disease. MATERIAL AND METHODS: A control group of 104 healthy subjects, and 272 patients with recurrent calcium oxalate stones were examined. Polymerase chain reaction based restriction endonuclease analysis was used to detect IL-18 polymorphisms. RESULTS: The patient and control groups differed significantly in genotypic expression of the IL-18 +105A/C polymorphism. The prevalence of the A/C + C/C genotypes in the patients was higher than that in the controls. The allelic frequency of IL-18 +105A/C differed significantly between the patients and the controls. The odds ratio (OR) of the A/C heterozygote of IL-18 +105A/C associated with urolithiasis was 2.772. The OR of the A/C + C/C genotypes of IL-18 +105A/C associated with urolithiasis was 3.097. The OR per copy of the C allele of IL-18 +105A/C associated with urolithiasis was 4.143. There were also significant differences in the prevalence of genotype IL-18 -137G/C polymorphisms between the patients and controls. The distribution of the G/G homozygote in the patients was higher than that in the controls. There was no significant difference in genotype and allelic frequency at the IL-18 -607C/A polymorphism between patients and control subjects. CONCLUSION: The results indicate that IL-18 +105A/C polymorphisms may play a role in the development of urolithiasis. PMID- 20017709 TI - Coeliac disease and reproductive disorders. AB - Reproductive problems, such as delayed menarche, amenorrhea, early menopause, infertility, impotence, hypogonadism, recurrent abortions, and low-birth-weight or preterm deliveries, are now known to be among the atypical symptoms of coeliac disease (CD). The pathogenesis of reproductive disorders in CD is unclear, but some hypotheses have been suggested, including autoimmunity and macro- and/or micronutrient deficiency. Recent investigations which have focused on tissue transglutaminase are promising with respect to the clarification of the mechanism of infertility and poor pregnancy outcomes in CD. In this review, the effects of CD on male and female reproductive disorders and pregnancy outcomes are discussed and the need for CD screening in the case of reproductive problems is emphasized. PMID- 20017710 TI - Is transcatheter arterial embolization a safer alternative than surgery when endoscopic therapy fails in bleeding duodenal ulcer? AB - OBJECTIVE: Emergency surgery after unsuccessful endoscopic therapy for bleeding duodenal ulcer has been reported to have a high mortality. Transcatheter arterial embolization (TAE) of the gastroduodenal artery is an alternative strategy when endoscopic therapy fails. This study is a retrospective analysis comparing these two treatment strategies. MATERIAL AND METHODS: Patients who underwent TAE (n = 24) or open surgery (n = 50) after unsuccessful endoscopic therapy for bleeding duodenal ulcers at two university hospitals between 2000 and 2007 were compared. Mortality, morbidity, length of hospital stay, age, number of endoscopic interventions and acute physiology and chronic health evaluation (APACHE) II score were evaluated. RESULTS: The groups were comparable concerning gender and length of hospital stay. The mean age (69.6 +/- 16.1 versus 61.9 +/- 14.1 years; P = 0.043), APACHE II score (17.0 +/- 5.1 versus 12.8 +/- 5.7; P = 0.004) and number of gastroscopies (P = 0.009) were significantly higher in the embolization group. Five patients (20.8%) died in the embolization group compared to 11 (22%) in the surgery group. However, mortality in high-risk patients (APACHE II score >or= 16.5) was lower in the TAE group (23.1% versus 50.0%; P = 0.236). Method related as well as other complications were not significantly different between the two groups. There was, however, a higher re-bleeding rate in the TAE group. CONCLUSIONS: TAE of the gastroduodenal artery appears to be a safe alternative when endoscopic therapy for bleeding duodenal ulcer fails, at least in high-risk patients. The role of TAE in low-risk patients with bleeding from duodenal ulcer needs to be defined by means of a prospective controlled trial. PMID- 20017711 TI - Confirmation of spectral jitter: a measured shift in the spectral distribution of intense pulsed light systems using a time-resolved spectrometer during exposure and increased fluence. AB - High quality intense pulsed light (IPL) systems can offer simple, safe and effective treatments for long-term hair reduction, skin rejuvenation and removal of benign vascular and pigmented lesions. Considerable differences in clinical efficacy and adverse effects have been recorded amongst different IPL systems despite comparable display settings. This study examines the variation in pulse structures exhibited by several popular professional IPL systems that can cause a spectral change within the broadband output depending on the pulse structure chosen by the system designers. A fast spectrometer was used to capture IPL spectral outputs. A spectral distribution shift that occurs both within a pulse and between pulses is clearly demonstrated and is more prominent with uncontrolled free discharge systems than with square pulsed technology, which provides a constant spectral distribution throughout the pulse duration. PMID- 20017712 TI - Modelling the interaction of ankle-foot orthosis and foot by finite element methods to design an optimized sole in steppage gait. AB - In this study, based on kinematics data in steppage gait, a finite element model of human gait with ankle-foot orthosis (AFO) has been proposed to optimize the role of AFO through minimizing stress in the patients' sole. The required kinetics data for the model were captured through a force plate and then analysed by 3D-DOCTOR and ANSYS software. In the proposed three-dimensional finite element model the transmitted tension in soft tissue and bones during gait was calculated. By changing the thickness and materials of different layers of sole in AFO the tension variations have been assessed. Unlike previous studies, the effect of orthosis on tension generated in bones and muscles has been dynamically and continuously modelled and the contact between AFO and sole has been considered in this model. By using the optimized sole the stress distribution has been changed by +50.38% in the forefoot, +15% in the midfoot and -37.79% in the hindfoot. The tension reduction in the sole can improve the effect of AFO during abnormal gait. It is possible to design each orthosis sole based on the kinetics data of each patient. PMID- 20017713 TI - An infant surgical table for laser photocoagulation: ergonomic improvement analysis. AB - Current methods of treatment for retinopathy of prematurity, using laser photocoagulation, require surgeons to assume awkward standing positions, which can result in occupational injury. A new infant surgical table was designed for improving this surgical procedure. To quantify its benefits, an ergonomic comparison of the standard and modified procedures was carried out, using specialized checklists, Nordic Musculoskeletal Questionnaires, and analysis of videotaped procedures using an Ovako Working Posture Analysing System method. Analysis of the typical laser photocoagulation procedure revealed a high risk for cumulative trauma disorders. The majority of the risk factors were lowered considerably with use of the new table. Improvement was largely due to the new table allowing seated postures during surgery, relieving muscular stress on the back, shoulders and legs. This study demonstrates risk reduction through engineering design of new medical devices, and illustrates how combining different assessment approaches can help evaluate ergonomic impact of medical technologies. PMID- 20017714 TI - International Society for Cell and Gene Therapy of Cancer 2009 Annual Meeting held in Cork, Ireland. AB - The International Society for Cell and Gene Therapy (ISCGT) of Cancer annual meeting was held from September 2 through September 4, 2009, in Cork, Ireland ( www.iscgt2009.com ). The conference was held in conjunction with the Irish Society for Gene and Cell Therapy third annual meeting, and brought together scientists and clinicians from around the world in a country developing its knowledge economy. Next year's ISCGT meeting will be held in Doha, the capital of Qatar ( www.iscgt.net ), from September 27 through September 29, 2010. PMID- 20017715 TI - Seasonal variations in human Toxoplasma infection in Serbia. AB - BACKGROUND: Seasonal variations in the occurrence of toxoplasmosis have been studied only sporadically. We thus examined the seasonal distribution of acute toxoplasmosis in Serbia during a 4-year period (2004-2008). MATERIALS AND METHODS: A total of 391 consecutive symptomatic (recent lymphadenopathy) and 715 asymptomatic (women tested for obstetric reasons) patients were tested for Toxoplasma immunoglobulin G (IgG) (including IgG avidity) and IgM antibodies. The distribution of patients with acute infection, and of all patients from both groups, was analyzed for seasonality. In addition, factors (including undercooked meat consumption, contact with cats and with soil) possibly contributing to seasonality were analyzed in patients with acute infection (cases) matched by age, sex, and time (month and year) of infection (symptomatic) or presentation (asymptomatic) with seronegative patients (controls). RESULTS: Acute toxoplasmosis was serologically (IgG avidity low, IgM positive) diagnosed in 39 (10.0%) symptomatic and 38 (5.3%) asymptomatic patients. In both groups, monthly distribution of acute infections showed significant (p < 0.0001) seasonality, which was related to the four seasons of the year (p < 0.0001). Importantly, the observed seasonality was not related to the distribution of all examined patients in either group (p < 0.001). In the symptomatic patients, acute infections occurred more often between October and March (p = 0.0486). Although more asymptomatic acute infections were diagnosed between February and July (p = 0.0037), low IgG avidity suggests that infection had occurred within the previous trimester (between November and April). Undercooked meat consumption was shown as a risk factor for symptomatic infection in the October-March period (odds ratio 7.67, 95% confidence interval 1.61-36.45). CONCLUSION: Seasonality patterns should be taken into account in the health education guidelines for the prevention of toxoplasmosis in pregnant women. PMID- 20017716 TI - Prevalence of Borrelia burgdorferi in Ixodes ricinus ticks in Belgrade area. AB - OBJECTIVE: Lyme borreliosis is vector-borne zoonosis. The causative agent of Lyme borreliosis is a spirochete of Borrelia burgdorferi (Bb) sensu lato complex, which is transmitted by ticks of the Ixodes ricinus complex. The aim of our paper is to estimate the prevalence of I. ricinus ticks, the level of their infectivity by Bb, and the prevalence of certain genospecies of Bb sensu lato in ixodide ticks inhabiting Belgrade. MATERIALS AND METHODS: An estimate of the tick population density was expressed by the value of flag/hour. For isolation and cultivation of Borrelia, selective Barbour-Stonner-Kelly II media was used. Typization of Borrelia was made by applying the species-specific polymerase chain reaction (PCR) and polymerase chain reaction-restriction fragment length polymorphism analysis. In statistical analysis, Chi(2) test was used. RESULTS: Values of flag/hour have varied in relation to year observed and type of habitat: The lowest values were recorded in the city parks (17.9). The values were higher in parks-woods (19.7 and 33.4, respectively). The highest values were detected in localities similar to wooded areas (48.0). The estimated average infestation of ticks with Bb was 21.9%, excluding statistically significant differences by years of investigation. We found the dominance of Borrelia afzelii (75%). Bb sensu stricto (22.2%) as well as Borrelia garinii (2.8%) was much less present. Statistically significant difference was established in the prevalence of the above-mentioned genospecies in relation to the examined localities. CONCLUSIONS: We have established the prevalence of all three genospecies in the city of Belgrade. Bb sensu lato was found, with the dominance of B. afzelii. PMID- 20017717 TI - Distribution of the Lyme disease spirochete Borrelia burgdorferi in naturally and experimentally infected western gray squirrels (Sciurus griseus). AB - The dynamics of Borrelia burgdorferi infections within its natural hosts are poorly understood. We necropsied four wild-caught western gray squirrels (Sciurus griseus) that were acquired during a previous study that evaluated the reservoir competence of this rodent for the Lyme disease spirochete. One animal was infected experimentally, whereas the others were infected in the wild before capture. To investigate dissemination of B. burgdorferi and concurrent histopathologic lesions in different tissues, blood specimens, synovial and cerebrospinal fluid, ear-punch biopsies, and diverse tissue samples from skin and various organs were taken and examined by culture, polymerase chain reaction, and histology. Borrelia-positive cultures were obtained from three of the squirrels, that is, from skin biopsies (7 of 20 samples), ear-punch biopsies (2 of 8), and one (1 of 5) lymph node. Sequencing of amplicons confirmed B. burgdorferi sensu stricto (s.s.) infection in 9 of 10 culture-positive samples and in DNA extracted from all 10 positive cultures. The experimentally infected squirrel yielded most of the positive samples. In contrast, bodily fluids, all other organ specimens from these animals, and all samples from one naturally infected squirrel were negative for Borrelia for both assays. None of the necropsied squirrels exhibited specific clinical signs associated with B. burgdorferi. Similarly, necropsy and histological examination of tissues indicated the presence of underlying infectious processes, none of which could be ascribed conclusively to B. burgdorferi infection. Based on these results, obtained from a small number of animals investigated at a single time point, we suggest that B. burgdorferi s.s. infection in S. griseus may result in rather localized dissemination of spirochetes, and that mild or nonclinical disease might be more common after several months of infection duration. Since spirochetes could be detected in squirrels 7-21 months postinfection, we conclude that S. griseus can infect Ixodes pacificus ticks with B. burgdorferi s.s. trans-seasonally. PMID- 20017718 TI - Detection of Bartonella tamiae DNA in ectoparasites from rodents in Thailand and their sequence similarity with bacterial cultures from Thai patients. AB - Ectoparasites, including chigger mites (genera Leptotrombidium, Schoengastia, and Blankarrtia) and one tick (genus Haemaphysalis) collected from wild-caught rodents in Thailand, were assessed for the presence of Bartonella DNA by using a polymerase chain reaction assay targeting the 16S-23S intergenic spacer region and citrate synthase gene (gltA). Of the 41 pooled samples tested, 34 were positive for Bartonella DNA. Sequence analysis demonstrated that DNA detected in 33 chigger mite pools and one tick pool was similar to Bartonella tamiae sequences previously isolated from three patients in Thailand. This is the first report of the detection of B. tamiae DNA in chigger mites; additional field and experimental investigations are required to determine the role of chigger mites as potential vectors of B. tamiae. PMID- 20017719 TI - Cliff swallows, swallow bugs, and West Nile virus: an unlikely transmission mechanism. AB - The cliff swallow (Petrochelidon pyrrhonota) could play an important role in the transmission of West Nile virus (WNV) because of its breeding ecology, reservoir competence status, and potentially high natural exposure rates. Cliff swallows nest within colonies and their nests are occupied year-round by swallow bugs (Oeciacus vicarius), hematophagus ectoparasites that feed primarily on cliff swallows. These parasites are likely exposed to WNV while feeding on infectious blood of nesting cliff swallow adults and nestlings and thus, if competent vectors, could contribute to seasonal elevations in WNV transmission. In addition, swallow bugs remain within nests year-round and therefore could provide a potential overwintering mechanism for WNV if persistently infected. To test the hypotheses that swallow bugs are competent vectors and become persistently infected with WNV, we experimentally inoculated cliff swallow nestlings, allowed swallow bugs to feed on these birds during the acute phase of infection, and then exposed naive cliff swallow nestlings to the same swallow bugs. In addition, a subset of swallow bugs that fed on infectious swallow nestlings was maintained through a simulated overwintering period. Although swallow bugs ingested infectious blood (up to 10(6.8) plaque-forming units of WNV/mL serum) and subsequently blood-fed on naive swallows, no WNV transmission was detected, and all bugs tested WNV negative after the simulated overwintering period. Although many ecologic scenarios exist beyond the present study, our results suggest that swallow bugs may be unlikely to serve as competent biological vectors for WNV during active transmission periods or to reinitiate seasonal transmission. PMID- 20017720 TI - Phosphoinositide 3-kinase gamma (PI3Kgamma) inhibitors for the treatment of inflammation and autoimmune disease. AB - Phosphoinositide 3-kinase gamma (PI3Kgamma) is a lipid kinase in leukocytes that generates phosphatidylinositol 3,4,5-trisphosphate to recruit and activate downstream signaling molecules. Distinct from other members in the PI3K family, PI3Kgamma is activated by G-protein coupled-receptors responding to chemotactic ligands. PI3Kgamma plays an important role in migration of both myeloid and lymphoid cells. It is also required for other leukocyte functions such as neutrophil oxidative burst, T cell proliferation and mast degranulation. Mice with PI3Kgamma inactivated by genetic or pharmacological approaches are protected from disease development in a number of inflammation and autoimmune disease models. The function of PI3Kgamma depends on its kinase activity and therefore it has been suggested by many reports that small molecules inhibiting its kinase activity could be promising for the treatment of inflammation and autoimmune diseases. Over the last five years, a number of pharmaceutical companies have reported a wide variety of PI3Kgamma inhibitors, of which several x-ray crystal structures with PI3Kgamma have been elucidated. The structural characteristics and selectivity profiles of these inhibitors, in particular thiazolidinones and 2 aminoheterocycles, and those disclosed in related patent applications are summarized in this review. PMID- 20017721 TI - Telomere maintenance as therapeutic target in embryonal tumours. AB - Embryonal tumours most commonly occur in the first few years of life and account for approximately 30% of childhood malignancies. Knowledge of these tumours' genetics has already impacted on their clinical management and further knowledge of their cellular immortalization will hopefully result in novel therapies. The ends of human chromosomes are capped and protected by telomeres; cellular replication, however, causes their loss. A critical length of telomere repeats is required to ensure proper telomere function and avoid the activation of DNA damage pathways that result in senescence and cell death. To proliferate beyond the senescence checkpoint, cells must restore their telomere length. Hence stabilization of telomere is an important step in cell immortalization and carcinogenesis. Telomere maintenance is evident in virtually all types of malignant cells, including embryonal tumours, where either a telomerase-dependent or alternative lengthening of telomeres (ALT) mechanism is employed in order to ensure their limitless replicative potential. For this reason effective strategies targeting telomere maintenance in cancer cells require a combination of telomerase and ALT inhibitors. In this review, we are giving an overview about telomere maintenance in childhood tumours and discussing its potential as a new therapeutic target. PMID- 20017722 TI - Targeted elimination of leukemia stem cells; a new therapeutic approach in hemato oncology. AB - Despite recent advances, treatment of leukemia is often not curative. New insights indicate that this may be attributable to a small population of therapy resistant malignant cells with self-renewal capacity and the ability to generate large numbers of more differentiated leukemia cells. These leukemia-initiating cells are commonly referred to as Leukemia Stem Cells (LSCs). LSCs are regarded as the root of leukemia origin and leukemia recurrence after seemingly successful therapy. Not surprisingly therefore, contemporary leukemia research has focused on ways to specifically eliminate LSCs, leading to the identification of several promising anti-LSC strategies. Firstly, LSCs may be eliminated by antibody- or ligand-based cell surface delivery of therapeutics such as naked antibodies, immunotoxins, and immunocytokines. This approach exploits LSC-associated surface antigens, such as CD33, CD44, CD96, CD123 and CLL-1 for LSC-selective therapy and aims to spare normal hematopoietic stem cells. A second strategy aims to disrupt the interactions between LSCs and their highly specialized niche. These interactions appear to be pivotal for maintenance of the stem cell-like characteristics of LSCs. A third strategy centers on the selective modulation of aberrantly activated signaling pathways central to LSC biology. A fourth strategy, dubbed 'epigenetic reprogramming', aims to selectively reverse epigenetic alterations that are implicated in ontogeny and maintenance of LSCs. In this review, we will discuss the rationale for these LSCs-targeted strategies and highlight recent advances that may ultimately help pave the way towards selective LSCs-elimination. PMID- 20017723 TI - Coenzyme Q10 in neuromuscular and neurodegenerative disorders. AB - Coenzyme Q10 (CoQ10, or ubiquinone) is an electron carrier of the mitochondrial respiratory chain (electron transport chain) with antioxidant properties. In view of the involvement of CoQ10 in oxidative phosphorylation and cellular antioxidant protection a deficiency in this quinone would be expected to contribute to disease pathophysiology by causing a failure in energy metabolism and antioxidant status. Indeed, a deficit in CoQ10 status has been determined in a number of neuromuscular and neurodegenerative disorders. Primary disorders of CoQ10 biosynthesis are potentially treatable conditions and therefore a high degree of clinical awareness about this condition is essential. A secondary loss of CoQ10 status following HMG-Coa reductase inhibitor (statins) treatment has be implicated in the pathophysiology of the myotoxicity associated with this pharmacotherapy. CoQ10 and its analogue, idebenone, have been widely used in the treatment of neurodegenerative and neuromuscular disorders. These compounds could potentially play a role in the treatment of mitochondrial disorders, Parkinson's disease, Huntington's disease, amyotrophic lateral sclerosis, Friedreich's ataxia, and other conditions which have been linked to mitochondrial dysfunction. This article reviews the physiological roles of CoQ10, as well as the rationale and the role in clinical practice of CoQ10 supplementation in different neurological and muscular diseases, from primary CoQ10 deficiency to neurodegenerative disorders. We also briefly report a case of the myopathic form of CoQ10 deficiency. PMID- 20017724 TI - The role of pemetrexed in lung cancer. PMID- 20017725 TI - Should adipokines be considered in the choice of the treatment of obesity-related health problems? AB - White adipose tissue (WAT) is an important endocrine organ that secretes approximately 30 biologically active peptides and proteins, collectively termed "adipokines". These are either produced exclusively by WAT (mainly adiponectin, leptin and resistin) or also by other tissues [e.g. tumour necrosis factor (TNF) alpha, interleukin (IL)-6, plasminogen activator inhibitor (PAI)-1, angiotensinogen]. Adipokines play a central role in body homeostasis including the regulation of food intake and energy balance, insulin action, lipid and glucose metabolism, angiogenesis and vascular remodelling, regulation of blood pressure and coagulation. Excess WAT, especially visceral obesity, is linked to obesity-related health problems through insulin resistance (IR) [leading to type 2 diabetes mellitus (T2DM)] and systemic low-grade inflammation [leading to cardiovascular disease (CVD)]. The adipokines are important mediators of these adverse effects. This review describes the role of proinflammatory adipokines in the pathogenesis of IR and of the chronic inflammatory state associated with visceral obesity. Moreover, it summarises treatment options for the normalisation of adipokine levels, which might confer an additional clinical benefit in the effort to prevent or treat obesity-related T2DM and CVD. PMID- 20017726 TI - Cannabinoids, opioids and MDMA: neuropsychological interactions related to addiction. AB - 3,4-methylenedioxymethamphetamine (MDMA, "ecstasy") is an amphetamine derivative with psychostimulant properties. This substance is widely used around the world by young adults in recreational settings. One of the most remarkable characteristic of ecstasy users is the concurrent consumption of several other drugs of abuse including psychostimulants, alcohol, tobacco, LSD, cannabis and opioids. This poly-drug pattern of use is now prompting research towards understanding how the combination of MDMA with cannabis and opioids could affect neuropsychobiological processes related to addiction. As with other drugs of abuse, behavioural evidence has been presented supporting the role of the endocannabinoid system as a modulator of the rewarding/reinforcing properties of MDMA. On the other hand, the neurochemical substrate for the complex interactions between the endocannabinoid system and MDMA is poorly understood. MDMA also modulates the activity of the dynorphinergic and enkephalinergic systems in several brain structures related to addiction, as it has been shown for other psychostimulants. The work regarding the contribution of micro- and delta-opioid receptors in the rewarding effects of MDMA shows differential results in pharmacological studies in rats, with respect to studies using knock-out mice. The present review describes the behavioural and neurochemical interactions between MDMA, cannabinoids and opioids with respect to addiction processes. PMID- 20017727 TI - Endogenous cannabinoid and opioid systems and their role in nicotine addiction. AB - Nicotine addiction is a complex behavioural alteration, in which many neuronal pathways and neurotransmitters are involved. For a long time, dopamine has been considered one of the most important neurotransmitters in mediating the rewarding effects of nicotine. In addition, a great amount of research suggests that the endogenous cannabinoid and opioid systems play an overall modulatory effect on the reward circuitry and participate in the addictive properties of most of the prototypical drugs of abuse. This review focuses on recent behavioural and biochemical data involving these systems in the different processes that contribute to tobacco addiction. A possible role for the endogenous cannabinoid and opioid systems in the rewarding properties of nicotine as well as in the development of nicotine physical dependence and relapse to nicotine-seeking behaviour will be examined. According to preclinical studies, clinical trials suggest that the manipulation of these systems with cannabinoid or opioid antagonists could be a potential therapeutical strategy for treating nicotine addiction. PMID- 20017728 TI - Opioids and cannabinoids interactions: involvement in pain management. AB - Among several pharmacological properties, analgesia is the most common feature shared by either opioid or cannabinoid systems. Cannabinoids and opioids are distinct drug classes that have been historically used separately or in combination to treat different pain states. Indeed, it is widely known that activation of either opioid or cannabinoid systems produce antinociceptive properties in different pain models. Moreover, several biochemical, molecular and pharmacological studies support the existence of reciprocal interactions between both systems, suggesting a common underlying mechanism. Further studies have demonstrated that the endogenous opioid system could be involved in cannabinoid antinociception and recent data have also provided evidence for a role of the endogenous cannabinoid system in opioid antinociception. These interactions may lead to additive or even synergistic antinociceptive effects, emphasizing their clinical relevance in humans in order to enhance analgesic effects with lower doses and consequently fewer undesirable side effects. Thus, the present review is focused on bidirectional interactions between opioids and cannabinoids and their potent repercussions on pain modulation. PMID- 20017729 TI - Cannabinoid-opioid interactions in drug discrimination and self-administration: effect of maternal, postnatal, adolescent and adult exposure to the drugs. AB - Cannabinoids and opioids are known to strictly interact in many physiological and pathological functions, including addiction. The endogenous opioid system is significantly influenced by maternal or perinatal cannabinoid exposure, major changes concerning operant behaviour in adult animals. Copious data suggests that adolescence is also a particularly sensitive period of life not only for the initiation of abusing illicit drugs, but also for the effects that these drugs exert on the neural circuitries leading to drug dependence. This paper examines the role played by the age of drug exposure in the susceptibility to discriminative and reinforcing effects of both cannabinoids and opioids. We first revisited evidence of alterations in the density and functionality of mu-opioid and CB1 cannabinoid receptors in reward-related brain regions caused by either maternal, postnatal, adolescent or adult exposure to opioids and cannabinoids. Then, we reviewed behavioural evidence of the long-term consequences of exposure to opioids and cannabinoids during gestation, postnatal period, adolescence or adulthood, focusing mostly on drug discrimination and self-administration studies. Overall, evidence confirms a neurobiological convergence of the cannabinoid and opioid systems that is manifest at both receptor and behavioural levels. Although discrepant results have been reported, some data support the gateway hypothesis that adolescent cannabis exposure contributes to greater opioid intake in adulthood. However, it should be kept into consideration that in humans genetic, environmental, and social factors could influence the direct neurobiological effects of early cannabis exposure to the progression to adult drug abuse. PMID- 20017730 TI - Cellular mechanisms underlying the interaction between cannabinoid and opioid system. AB - Recently, the presence of functional interaction between the opioid and cannabinoid system has been shown in various pharmacological responses. Although there is an increasing interest for the feasible therapeutic application of a co administration of cannabinoids and opioids in some disorders (i.e. to manage pain, to modulate immune system and emotions) and the combined use of the two drugs by drug abusers is becoming largely diffuse, only few papers focused on cellular and molecular mechanisms underlying this interaction. This review updates the biochemical and molecular underpinnings of opioid and cannabinoid interaction, both within the central nervous system and periphery. The most convincing theory for the explanation of this reciprocal interaction involves (i) the release of opioid peptides by cannabinoids or endocannabinoids by opioids, (ii) the existence of a direct receptor-receptor interaction when the receptors are co-expressed in the same cells, and (iii) the interaction of their intracellular pathways. Finally, the cannabinoid/opioid interaction might be different in the brain rewarding networks and in those accounting for other pharmacological effects (antinociception, modulation of emotionality and cognitive behavior), as well as between the central nervous system and periphery. Further insights about the cannabinoid/opioid interaction could pave the way for new and promising therapeutic approaches. PMID- 20017731 TI - Antifungal curcumin induces reactive oxygen species and triggers an early apoptosis but prevents hyphae development by targeting the global repressor TUP1 in Candida albicans. AB - In the present study, we have investigated the antifungal effects of a natural polyphenol, CUR (curcumin), against albicans and non-albicans species of Candida and have shown its ability to inhibit the growth of all the tested strains. The inhibitory effects of CUR were independent of the status of the multidrug efflux pump proteins belonging to either ABC transporter (ATP-binding cassette transporter) or MFS (major facilitator) superfamilies of transporters. By using a systemic murine model of infection, we established that CUR and piperine, when administered together, caused a significant fungal load reduction (1.4log10) in kidneys of Swiss mice. Additionally, CUR raised the levels of ROS (reactive oxygen species), which, as revealed by annexin V-FITC labelling, triggered early apoptosis in Candida cells. Coincident with the raised ROS levels, mRNAs of tested oxidative stress-related genes [CAP1 (Candida albicans AP-1), CaIPF7817 (putative NADH-dependent flavin oxidoreductase), SOD2 (superoxide dismutase 2), GRP2 (NADPH-dependent methyl glyoxal reductase) and CAT1 (catalase 1)] were also elevated. The growth inhibitory effects of CUR could be reversed by the addition of natural and synthetic antioxidants. Notably, independent of ROS status, polyphenol CUR prevented hyphae development in both liquid and solid hypha inducing media by targeting the global suppressor TUP1 (thymidine uptake 1). Taken together, our results provide the first evidence that CUR acts as an antifungal agent, via generation of oxidative stress, and inhibits hyphae development by targeting TUP1. PMID- 20017733 TI - Secretory vesicles transiently dock and fuse at the porosome to discharge contents during cell secretion. AB - In contrast with the observation in electron micrographs of partially empty vesicles in cells following secretion, it has been believed since the 1950s that during cell secretion, secretory vesicles completely merge at the cell plasma membrane, resulting in the diffusion of intravesicular contents to the cell exterior and the compensatory retrieval of the excess membrane by endocytosis. In the interim, a large body of work has been published arguing both for and against the complete merger of secretory vesicle membrane at the cell plasma membrane during secretion. The only definitive determination of the mechanism of cell secretion remained in its direct observation at nanometre resolution in live cells. In the past decade, this finally became a reality through the power and scope of the atomic force microscope, which has made it possible to resolve a major conundrum in cell biology. This paradigm shift in our understanding of cell secretion is briefly outlined here. PMID- 20017734 TI - Discovery of functional motifs in h-regions of trypanosome signal sequences. AB - N-terminal signal peptides direct secretory proteins into the ER (endoplasmic reticulum) of eukaryotes or the periplasmic space of prokaryotes. A hydrophobic core (h-region) is important for signal sequence function; however, the mechanism of h-region action is not resolved. To gain new insight into signal sequences, bioinformatic analysis of h-regions from humans, Saccharomyces cerevisiae, Trypanosoma brucei and Escherichia coli was performed. Each species contains a unique set of peptide motifs (h-motifs) characterized by identity components (i.e. sequence of conserved amino acids) joined by spacers. Human h-motifs have four identity components, whereas those from the other species utilize three identity components. Example of h-motifs are human Hs3 {L-x(2)-[AGILPV]-L-x(0,2) L}, S. cerevisiae Sc1 [L-x(0,2)-S-x(0,3)-A], T. brucei Tb2 {L-x(1,2)-L-[AILV]} and E. coli Ec1 [A-x(0,2)-L-x(0,3)-A]. The physiological relevance of h-motifs was tested with a T. brucei microsomal system for translocation of a VSG (variant surface glycoprotein)-117 signal peptide. Disruption of h-motifs by scrambling of sequences in h-regions produced defective signal peptides, although the hydrophobicity of the peptide was not altered. We conclude that: (i) h-regions harbour h-motifs, and are not random hydrophobic amino acids; (ii) h-regions from different species contain unique sets of h-motifs; and (iii) h-motifs contribute to the biological activity of ER signal peptides. h-Regions are 'scaffolds' in which functional h-motifs are embedded. A hypothetical model for h-motif interactions with a Sec61p protein translocon is presented. PMID- 20017732 TI - The multiple roles of myelin protein genes during the development of the oligodendrocyte. AB - It has become clear that the products of several of the earliest identified myelin protein genes perform functions that extend beyond the myelin sheath. Interestingly, these myelin proteins, which comprise proteolipid protein, 2',3' cyclic nucleotide 3'-phosphodiesterase and the classic and golli MBPs (myelin basic proteins), play important roles during different stages of oligodendroglial development. These non-myelin-related functions are varied and include roles in the regulation of process outgrowth, migration, RNA transport, oligodendrocyte survival and ion channel modulation. However, despite the wide variety of cellular functions performed by the different myelin genes, the route by which they achieve these many functions seems to converge upon a common mechanism involving Ca(2+) regulation, cytoskeletal rearrangements and signal transduction. In the present review, the newly emerging functions of these myelin proteins will be described, and these will then be discussed in the context of their contribution to oligodendroglial development. PMID- 20017735 TI - Epidemiology of exfoliation syndrome in the Reykjavik Eye Study. AB - Exfoliation syndrome (XFS) is a major risk factor for glaucoma. It is characterized by a pathological accumulation of polymorphic fibrillar material in the anterior segment of the eye. It is likely that the increase in intraocular pressure (IOP) seen in XFS patients is at least in part because of flakes of material clogging up the trabecular meshwork, and thereby increasing the resistance to outflow and increasing IOP. XFS glaucoma progresses more rapidly, is more resistant to medical treatment and has worse prognosis than other glaucomas. The prevalence of XFS has been found to vary greatly between different studies, raising the possibility of racial and/or environmental modulators. XFS has also been linked to other changes in ophthalmological structures such as; changes in central corneal thickness (CCT), steeper corneal curvature (CC) and nuclear lens opacifications. Some studies have found XFS to be associated with systemic diseases, mostly cardiovascular and cerebrovascular. Exposure to ultra violet (UV) light has also been investigated as a possible culprit, along with several other plausible factors. The aim of the present study was to determine the prevalence and 5-year incidence of XFS, to establish possible risk factors and/or concomitant symptoms and finally to investigate the relationship between XFS and glaucomatous changes. The Reykjavik Eye Study (RES) is a prospective study based on a random sample from the Icelandic national population sample. The baseline examination was performed in the autumn of 1996, when 1045 persons older than 50 years participated. Of these, 846 (88.2% of survivors) participated in a follow-up 5 years later. All participants went through a standard examination protocol, and answered a comprehensive questionnaire on health and life style. In the prevalence study, XFS was found in 10.7% of subjects, more frequently in women and older persons. Five years later, a further 5.2% of those that participated in the follow-up study and had no signs of XFS at baseline were diagnosed having XFS. We found a strong correlation between IOP and XFS. No difference was found in the anterior segment parameters measured, but there was a significant loss of neural tissue in the XFS as demonstrated by measurements of cup/disk ratio. In conclusion, we find XFS to be frequent among Icelanders, increasing with age and more in women. Our diagnostic criteria are reliable over time. We have also identified possible risk factors that point to a role of antioxidants in the development of XFS. We find changes in corneal curvature and thickness more related to age than XFS. PMID- 20017737 TI - Abstracts of the VI National Congress of Pharmacology. October 1-4, 2009. Varna, Bulgaria. PMID- 20017740 TI - The cutting edge of transplant surgery: committed to leading the way in the 21st century. Abstracts of the ASTS (American Society of Transplant Surgeons) 10th Annual State of the Art Winter Symposium. January 15-17, 2010. PMID- 20017741 TI - The perils of reducing humans to objects of research. PMID- 20017742 TI - Research on prisoners - a comparison between the IOM Committee recommendations (2006) and European regulations. AB - The Institute of Medicine (IOM) Committee on Ethical Considerations for Revisions to DHHS Regulations for Protection of Prisoners Involved in Research published its report in 2006. It was charged with developing an ethical framework for the conduct of research with prisoners and identifying the safeguards and conditions necessary to ensure that research with prisoners is conducted ethically. The recommendations contained in the IOM report differ from current European regulations in several ways, some being more restrictive and some less so. For example, the IOM report suggests limiting the percentage of prisoners that should be involved in a biomedical study to 50%, a limit that does not exist in Europe. However, the report does not specifically advise against research without a direct benefit to an individual prisoner: the European regulations are more restrictive than the IOM committee recommendations in this respect. The definition of minimal risk varies, as well as the proposed role of the minimal risk requirement and of the principle of subsidiarity (research that can only be done effectively in prisons). The IOM report proposes a number of thoughtful suggestions, which it would be beneficial to implement everywhere, such as registers of research on prisoners. The European regulations offer pragmatic solutions to several thorny issues. In summary, the IOM committee report represents an admirable effort to tackle the present inconsistencies and deficiencies of federal regulations in the US on research on prisoners (45 CFR 46 Subpart C). Nonetheless, before acting on the recommendations, US regulators might consider revisiting international guidelines such as those published by the Council for International Organizations of Medical Science (CIOMS) and the Declaration of Helsinki. PMID- 20017743 TI - Against risk-benefit review of prisoner research. AB - The 2006 Institute of Medicine (IOM) report, 'Ethical Considerations for Research Involving Prisoners', recommended five main changes to current US Common Rule regulations on prisoner research. Their third recommendation was to shift from a category-based to a risk-benefit approach to research review, similar to current guidelines on pediatric research. However, prisoners are not children, so risk benefit constraints on prisoner research must be justified in a different way from those on pediatric research. In this paper I argue that additional risk benefit constraints on prisoner research are unnecessary: the current Common Rule regulations, omitting category-based restrictions but conjoined with the IOM report's other four main recommendations, ensure that prisoner research is as ethical as non-prisoner research is. I explain why four problems which which may be more prevalent in prisons and which risk-benefit constraints may seem to address - coercion, undue inducements, exploitation, and protection from harm - are in fact not solved by adding further risk-benefit constraints on prisoner research. PMID- 20017744 TI - Prisoner research - looking back or looking forward? AB - Much has been written about prisoner research and the controversies surrounding prisoners as human subjects. The Institute of Medicine recently released a report addressing some of these issues. This report, which generated further controversy, needs to be fully discussed in the literature and certain aspects are examined in this work. Further, in the body of literature there has been little acknowledgement of the concept of the right of prisoners to be involved in research. This needs to be pursued from an ethical perspective and eventually a legal one. This paper explores that concept and documents some facilities in which a prisoner's right to research has occurred. PMID- 20017745 TI - Detained and drugged: a brief overview of the use of pharmaceuticals for the interrogation of suspects, prisoners, patients, and POWs in the US. AB - Using medical literature citations, Congressional hearings, and declassified documents this paper examines the uses of pharmaceuticals in the interrogation of vulnerable populations. From the use of IV relaxants on criminal suspects during the 1920s to the Global War on Terror, the nexus of drugs, testing, and interrogations will be explored in both the domestic and international contexts. PMID- 20017746 TI - Ethical issues in forensic psychiatric research on mentally disordered offenders. AB - This paper analyses ethical issues in forensic psychiatric research on mentally disordered offenders, especially those detained in the psychiatric treatment system. The idea of a 'dual role' dilemma afflicting forensic psychiatry is more complicated than acknowledged. Our suggestion acknowledges the good of criminal law and crime prevention as a part that should be balanced against familiar research ethical considerations. Research aiming at improvements of criminal justice and treatment is a societal priority, and the total benefit of studies has to be balanced against the risks for research subjects inferred by almost all systematic studies. Direct substantial risks must be balanced by health benefits, and normal informed consent requirements apply. When direct risks are slight, as in register-based epidemiology, lack of consent may be counter-balanced by special measures to protect integrity and the general benefit of better understanding of susceptibility, treatment and prevention. Special requirements on consent procedures in the forensic psychiatric context are suggested, and the issue of the relation between decision competence and legal accountability is found to be in need of further study. The major ethical hazard in forensic psychiatric research connects to the role of researchers as assessors and consultants in a society entertaining strong prejudices against mentally disordered offenders. PMID- 20017747 TI - "Increasing our understanding of migraine to enhance treatment outcomes"- symposium. Foreword. PMID- 20017748 TI - Modulation of trigeminovascular processing: novel insights into primary headache disorders. AB - Dysfunction of the descending circuitry that modulates nociceptive processing is thought to facilitate primary headache disorders such as migraine and cluster headache, either by reducing inhibition or increasing facilitation of neural signals. The identification of novel neurotransmitter systems (orexinergic, cannabinoid and dopaminergic) within this descending pathway and their interaction with 5-HT(1B/1D) (triptan) receptors provides important clues about several of the risk factors and comorbidities associated with primary headache disorders and indicates that triptans exert central as well as peripheral activity. Current research supports the early treatment of primary headache disorders in order to prevent structural changes in the brain and to minimize dysfunction in the descending modulation of pain control. PMID- 20017749 TI - Review of comorbidities and risk factors for the development of migraine complications (infarct and chronic migraine). AB - A number of comorbid disorders, behavioural traits and associated risk factors in patients with migraine are known to increase the risk of complications such as ischaemic vascular events and chronic migraine, a syndrome that is more disabling and resistant to treatment with acute and preventative medications than episodic migraine. Reduction of cardiovascular risk factors, smoking cessation and use of non-oestrogen-containing oral contraceptives in female patients are beneficial strategies to reduce the risk of ischaemic events in patients with migraine (especially those with aura). Attack frequency, acute medication overuse, obesity and coexisting depression and anxiety disorders are particularly strong but potentially modifiable independent risk factors for progression to chronic migraine. Identifying and managing comorbidities and associated risk factors for complications of migraine are likely to require an integrated disease management strategy involving several disciplines and allied health services. Such a disease oriented model of care may potentially interrupt the cycle of progression and disability and improve quality of life for patients with migraine. PMID- 20017750 TI - Early treatment of acute migraine: new evidence of benefits. AB - The current management approach to migraine headaches advocates use of triptan medications early in the course of an attack while pain is still mild, rather than waiting to treat the pain when it has progressed to moderate-severe. Recently, strong new evidence for the benefits of early intervention has become available. The AEGIS, AIMS and AwM studies of almotriptan in patients with migraine indicate that earlier treatment initiation and lower pain intensity at the time of treatment are important predictors of enhanced therapeutic outcomes. The opportunity to treat early exists for about 50% of all migraine attacks, which offers considerable scope for improving migraine management. Importantly, treating pain early and before it has progressed beyond 'mild' meets many of the expectations patients have of their migraine treatment. It is believed that consistent, positive outcomes may assist in overcoming the various physician- and patient-perceived barriers to adoption of this beneficial treatment strategy. PMID- 20017751 TI - Question & answer session. PMID- 20017752 TI - Prophylactic therapy with Fibrogammin P is associated with a decreased incidence of bleeding episodes: a retrospective study. AB - SUMMARY: Congenital factor XIII (FXIII) deficiency is an extremely rare, yet potentially life-threatening, bleeding disorder, with a 30% rate of spontaneous intracranial haemorrhage. Routine prophylactic management is recommended for all individuals with clinically relevant (FXIII) deficiency and for all symptomatic individuals with congenital factor deficiency. Fibrogammin P is a purified, pasteurized concentrate of FXIII that appears to carry negligible risk of viral transmission, unlike other unprocessed products containing FXIII. An ongoing Phase II/III study of Fibrogammin P in patients with congenital FXIII deficiency is being conducted to evaluate the prophylactic efficacy and long-term safety of this product. Using retrospective chart review data from subjects enrolled in the Phase II/III study, the current analysis was designed to compare spontaneous bleed-event rates prior to and after the initiation of Fibrogammin P prophylaxis. Seven subjects were evaluable for comparison, having received no other prophylactic FXIII-containing product during the 24 months prior to study entry. The mean annual number of spontaneous bleeds was 2.5 events per year prior to Fibrogammin P prophylaxis and 0.2 events per year during Fibrogammin P prophylaxis (P = 0.01). Patients reported no severe bleeds during Fibrogammin P therapy. This small sample supports a consistent and clinically meaningful reduction in spontaneous bleeding with prophylactic use of Fibrogammin P. PMID- 20017754 TI - Harrow: new Drosophila hAT transposons involved in horizontal transfer. AB - In this study we characterize the transposable elements harrow, which belong to the hAT superfamily of DNA transposons. Searches for harrow sequences were performed in 65 Drosophilidae species, mainly representing Neotropical and cosmopolitan groups from the genus Drosophila. The nucleotide divergence among elements found in these species suggests that harrow sequences could be clustered in a subfamily. The patchy distribution throughout the genus Drosophila and the high similarity presented between all harrow sequences indicate that horizontal transfer could play a major role in the evolution of harrow elements. The results obtained suggest an evolutionary scenario in which harrow would have undergone multiple horizontal transfer events in the Neotropics, involving D. tripuncatata, D. mojavensis (Subgenus Drosophila) and several species of the willistoni and saltans groups (subgenus Sophophora). PMID- 20017755 TI - Insect intestinal mucins and serine proteases associated with the peritrophic matrix from feeding, starved and moulting Mamestra configurata larvae. AB - Insect intestinal mucins (McIIM2-4) expressed in the midgut of feeding, starved and moulting Mamestra configurata larvae were identified. McIIM2 and McIIM4 were associated with the peritrophic matrix (PM). PMs from feeding and starved larvae were translucent and contained organized chitin bundles perpendicular to their long axis, whereas PM from moulting larvae consisted of an inner opaque mass surrounded by an outer translucent sleeve. Serine protease genes (McSP1, McSP2, McSP25 and McSP29) were also expressed in these larvae and several serine proteases were associated with the PM. Serine protease activity was also detected in the midgut of feeding, starved and moulting larvae. PMID- 20017753 TI - Analysis of Rickettsia typhi-infected and uninfected cat flea (Ctenocephalides felis) midgut cDNA libraries: deciphering molecular pathways involved in host response to R. typhi infection. AB - Murine typhus is a flea-borne febrile illness that is caused by the obligate intracellular bacterium, Rickettsia typhi. The cat flea, Ctenocephalides felis, acquires R. typhi by imbibing a bloodmeal from a rickettsemic vertebrate host. To explore which transcripts are expressed in the midgut in response to challenge with R. typhi, cDNA libraries of R. typhi-infected and uninfected midguts of C. felis were constructed. In this study, we examined midgut transcript levels for select C. felis serine proteases, GTPases and defence response genes, all thought to be involved in the fleas response to feeding or infection. An increase in gene expression was observed for the serine protease inhibitors and vesicular trafficking proteins in response to feeding. In addition, R. typhi infection resulted in an increase in gene expression for the chymotrypsin and rab5 that we studied. Interestingly, R. typhi infection had little effect on expression of any of the defence response genes that we studied. We are unsure as to the physiological significance of these gene expression profiles and are currently investigating their potential roles as it pertains to R. typhi infection. To our knowledge, this is the first report of differential expression of flea transcripts in response to infection with R. typhi. PMID- 20017756 TI - piggyBac-like elements in the pink bollworm, Pectinophora gossypiella. AB - A transgenic line of the pink bollworm, Pectinophora gossypiella, a key lepidopteran cotton pest, was generated previously using the piggyBac transposon IFP2 from Trichoplusia ni. Here we identified an endogenous piggyBac-like element (PLE), designated as PgPLE1, in the pink bollworm. A putatively intact copy of PgPLE1 (PgPLE1.1) presents the canonical features of PLE: inverted terminal repeats with three C/G residues at the extreme ends, inverted subterminal repeats, TTAA target site and an open reading frame encoding transposase with 68% similarity to IFP2. Vectorette PCR revealed large variation in the insertion sites of PgPLE1 amongst worldwide populations, indicating the potential mobility of PgPLE1. The PgPLE1 was undetectable in the genome of Pectinophora endema, implying the recent invasion of PgPLE1 after the divergence of these two closely related species. PMID- 20017757 TI - Abstracts of the 12th Annual Meeting of the European Society for Sexual Medicine. Lyon, France. November 15-18, 2009. PMID- 20017758 TI - Plasma arginine vasopressin concentration in horses undergoing surgery for colic. AB - OBJECTIVE: To determine if horses before undergoing anesthesia for surgical correction of colic would have lower plasma arginine vasopressin (AVP) concentrations than healthy horses undergoing anesthesia for arthroscopic surgery, and would not increase their plasma AVP concentrations in response to anesthesia and surgery. DESIGN: Prospective clinical study. SETTING: University teaching hospital. ANIMALS: Fourteen horses with colic and 8 healthy horses. INTERVENTIONS: Horses with colic underwent anesthesia and surgery for alleviation of colic, and healthy horses underwent anesthesia and surgery for arthroscopy. MEASUREMENTS AND MAIN RESULTS: Plasma AVP was measured perioperatively in horses with colic and in healthy horses. Before anesthesia, and 30 and 60 minutes after induction, horses with colic had greater median plasma AVP concentrations than control horses (P or =2401 CD38 ABC and > or =85% CD38/CD8 cut-off points, accurately discriminates responders versus non-responders, both measures resulting in 75.0% (CI 42.8-94.5) sensitivity (identification of non-responder) and 93.8% (CI 69.8-99.8) specificity (identification of responder), when considered as single assays. The association '> or =2401 CD38 ABC or > or =85% CD38/CD8' improved sensitivity to 83.3% (CI 51.6-97.9), while the association '<2401 CD38ABC (or <85% CD38/CD8) and lymphoproliferative response positive to > or =2 tested organisms' improved specificity to 100% (CI 79.4-100). In conclusions, CD38 expression and mycotic antigen-specific T-cell proliferation may be used as additional parameters to existing criteria to evaluate antiretroviral response in immuno-virological discordant patients. PMID- 20017810 TI - Case-control study of a HLA-G 14-bp insertion-deletion polymorphism in women with recurrent miscarriages. AB - Human leucocyte antigen (HLA)-G is an important molecule for maintaining an immunotolerant foetal-maternal relationship. A 14-bp insertion (+14-bp)/deletion (-14-bp) polymorphism in exon 8 of the 3' untranslated region of the HLA-G gene has been proposed to be associated with HLA-G mRNA stability and the expression of HLA-G. This might play an immunomodulatory role in human pregnancy. In this study, a total of 526 Chinese women were genotyped for the +14-bp/-14-bp polymorphism, including 137 who had recurrent miscarriages (three miscarriages: 86, four or more miscarriages: 51), 138 women who had experienced two abortions and 251 women with normal fertility as controls. The +14-bp homozygote sequence was more prominent among those with recurrent miscarriages (three or more recurrent miscarriages) in contrast to fertile control women. Significant difference was observed in the distribution of +14-bp/+14-bp genotype between controls and the recurrent abortions group with four or more abortions. A 14-bp insertion/deletion polymorphism in exon 8 has a possible role in HLA-G expression in certain cases of recurrent miscarriage. However, additional studies are needed in this regard. PMID- 20017811 TI - Application of an ELISA-elution assay to dissociate digoxin-antibody complexes in immunoaffinity chromatography. AB - In this study, we used a modified enzyme-linked immunosorbent assay (ELISA) elution technique as a screening tool to select specific elution conditions. We examined 12 different elution conditions for the removal of antibodies from a complex on an ELISA plate; 0.2 mol/l glycine-HCl (pH 2.5), 1.0 mol/l acetic acid (pH 2.5), 25% methanol (pH 2.5) and 3 mol/l NaSCN showed a higher elution efficiency. We conducted affinity chromatography with these four conditions for the purification of anti-digoxin antibodies from hyperimmune sera with a digoxin specific column using omega-aminoalkyl derivatives of Sepharose 4B, whose elution efficiency was similar to that of ELISA. We also monitored the relative specific activities during elution from the digoxin-specific column. The optimum, general purpose dissociation reagent for this immunoaffinity system was identified as 25% methanol (pH 2.5) with an elution efficiency and relative specific activity of 88.40% and 62.25%, respectively. The high purity of the purified antibodies was demonstrated with sodium dodecyl sulphate-polyacrylamide gel electrophoresis. PMID- 20017813 TI - The sedative effects of low-dose medetomidine and butorphanol alone and in combination intravenously in dogs. AB - OBJECTIVE: To evaluate the sedative effects of intravenous (IV) medetomidine (1 microg kg(-1)) and butorphanol (0.1 mg kg(-1)) alone and in combination in dogs. STUDY DESIGN: Prospective, blinded, randomized clinical trial. ANIMALS: Sixty healthy (American Society of Anesthesiologists I) dogs, aged 6.2 +/- 3.2 years and body mass 26 +/- 12.5 kg. METHODS: Dogs were assigned to four groups: Group S (sodium chloride 0.9% IV), Group B (butorphanol IV), Group M (medetomidine IV) and Group MB (medetomidine and butorphanol IV). The same clinician assessed sedation before and 12 minutes after administration using a numerical scoring system in which 19 represented maximum sedation. Heart rate (HR), respiratory rate, pulse quality, capillary refill time and rectal temperature were recorded after each sedation score assessment. Sedation scores, sedation score difference (score after minus score before administration) and patient variables were compared using one-way anova for normally distributed variables and Kruskal Wallis test for variables with skewed distributions and/or unequal variances. Where significance was found, further evaluation used Bonferroni multiple comparisons for pair-wise testing. RESULTS: Breed, sex, neuter status, age and body mass did not differ between groups. Sedation scores before substance administration were similar between groups (p = 0.2). Sedation scores after sedation were significantly higher in Group MB (mean 9.5 +/- SD 5.5) than in group S (2.5 +/- 1.8) (p < 0.001), group M (3.1 +/- 2.5) (p < 0.001) and group B (3.7 +/- 2.0) (p = 0.003). Sedation score difference was significantly higher in Group MB [7 (0-13)] than in Group S [0 (-1 to 4)] (p < 0.001) and Group M [0 (0 6)] (p < 0.001). HR decreased significantly in Groups M and MB compared with Group S (p < 0.05). CONCLUSION AND CLINICAL RELEVANCE: Low-dose medetomidine 1 microg kg(-1) IV combined with butorphanol 0.1 mg kg(-1) IV produced more sedation than medetomidine or butorphanol alone. HR was significantly decreased in both medetomidine groups. PMID- 20017814 TI - Alfaxalone induction dose following administration of medetomidine and butorphanol in the dog. AB - OBJECTIVE: To determine in dogs the effects of medetomidine and butorphanol, alone and in combination, on the induction dose of alfaxalone and to describe the induction and intubation conditions. STUDY DESIGN: Prospective, randomized, blinded clinical trial. ANIMALS: Eighty-five client-owned dogs (ASA 1 or 2). METHODS: Subjects were block randomized to treatment group according to temperament. The treatment groups were: medetomidine 4 microg kg(-1) (M), butorphanol 0.1 mg kg(-1) (B), or a combination of both (MB), all administered intramuscularly. After 30 minutes, a sedation score was assigned, and alfaxalone 0.5 mg kg(-1) was administered intravenously over 60 seconds by an observer who was unaware of treatment group. Tracheal intubation conditions were assessed and, if tracheal intubation was not possible after 20 seconds, further boluses of 0.2 mg kg(-1) were given every 20 seconds until intubation was achieved. Induction dose and adverse events (sneezing, twitching, paddling, excitement, apnoea and cyanosis) were recorded; induction quality and intubation conditions were scored and recorded. RESULTS: The mean dose of alfaxalone required for induction was similar for groups M and B: 1.2 +/- 0.4 mg kg(-1). The mean dose requirement for group MB (0.8 +/- 0.3 mg kg(-1)) was lower than groups M and B (p < 0.0001). Induction dose was not influenced by temperament or level of sedation. Induction and intubation scores did not differ between treatment groups. Adverse events were noted in 16 dogs; there was no association with treatment group, temperament or level of sedation. CONCLUSIONS AND CLINICAL RELEVANCE: Medetomidine and butorphanol administered in combination reduce the anaesthetic induction dose of alfaxalone compared to either agent alone. This difference should be taken into account when using this combination of drugs in a clinical setting. PMID- 20017815 TI - Electroencephalography during ovariohysterectomy in rats anaesthetized with halothane. AB - OBJECTIVE: To evaluate electroencephalographic (EEG) changes during ovariohysterectomy (OVH) in rats anaesthetized with halothane, and modification of the EEG changes by the co-administration of fentanyl, ketamine or thiopental. STUDY DESIGN: Prospective, randomized, blinded controlled study. ANIMALS: Sixty adult female Sprague-Dawley rats. METHODS: Anaesthesia was induced and maintained with halothane [Fe'HAL 0.95 +/- 0.05%]. The electroencephalogram was recorded continuously from the left and right primary somatosensory cortices. Rats were randomly divided into four groups and a rapid IV infusion of physiological saline, thiopental, ketamine or fentanyl was administered. OVH started 10 minutes after drug administration. Blood samples, for assay of plasma drug concentrations, were collected 5 minutes after administration and at the end of surgery. Electroencephalograph descriptors median frequency (F50), spectral edge frequency 95% (F95) and total power (P(tot)), recorded during non-surgical baseline periods, were compared with those recorded during defined surgical periods; skin incision, right and left ovarian pedicle ligation; cervical ligation. Plasma drug concentrations were measured using high performance lipid chromatography. RESULTS: Although a large number of statistical differences in EEG data were observed, these generally represented a reduction in F50 or F95 throughout the experimental recording period and were similar between groups. A significant reduction in P(tot) occurred during ligation of the ovaries and cervix in the control group compared with other recording periods. The co administration of ketamine, fentanyl and thiopental obtunded the reduction in P(tot). CONCLUSIONS AND CLINICAL RELEVANCE: Electroencephalographic changes, in the control group, mimicked changes reported in other studies using the minimal anaesthesia model. However, the stability in F50 during the surgical period compared with the baseline period indicated that OVH is an unsuitable surgical stimulus to investigate EEG changes with noxious stimulation. This may be attributed to the relatively prolonged duration of this surgical procedure and the primarily visceral afferent sensory innervation of the genital tract. PMID- 20017816 TI - The bispectral index during recovery from halothane and sevoflurane anaesthesia in horses. AB - OBJECTIVE: To record the bispectral index (BIS) when horses moved during either halothane or sevoflurane anaesthesia and when they made volitional movements during recovery from these anaesthetics. STUDY DESIGN: Randomized prospective clinical study. ANIMALS: Twenty-five client-owned horses undergoing surgery aged 8.8 (+/- 5.3; 1-19) years (mean +/- SD; range). METHODS: Baseline BIS values were recorded before pre-anaesthetic medication (BIS(B)) and during anaesthesia (BIS(A)) maintained with halothane (group H; n = 12) or sevoflurane (group S; n =13) at approximately 0.8-0.9 x minimum alveolar concentrations (MAC). Bispectral indices were recorded during the surgery when unexpected movement occurred (BIS(MA)), during recovery when the first movement convincingly associated with consciousness was observed (BIS(M1)) and once sternal recumbency was achieved (BIS(ST)). RESULTS: No significant difference in BIS(M1) was found between halothane- (85 +/- 7; 75-93) and sevoflurane- (87 +/- 10; 70-98) anaesthetized horses although BIS(A) was significantly (p = 0.0002) lower in group S (62 +/- 7; 53-72) than group H (74 +/- 7; 60-84). Differences between BIS(M1) and BIS(A) were significant in sevoflurane (p = 0.00001) and halothane recipients (p = 0.002) but were greater in group S (25 +/- 9; 4-38) compared with group H (12 +/- 10; -9-25). In six of eight horses, BIS(MA) values ranged between those recorded during anaesthesia and at first movement. CONCLUSIONS AND CLINICAL RELEVANCE: Bispectral indices appear to approximate levels of unconsciousness, suggesting that monitoring the BIS may assist equine anaesthesia. However, it does not predict intra-operative movement. PMID- 20017817 TI - Effects of opioids and anesthetic drugs on body temperature in cats. AB - OBJECTIVE: To determine which class of opioid alone or in conjunction with other anesthetic drugs causes post-anesthetic hyperthermia in cats. STUDY DESIGN: Prospective, randomized, crossover study. ANIMALS: Eight adult, healthy, cats (four spayed females and four castrated males weighing 3.8 +/- 0.6 kg). METHODS: Each cat was instrumented with a wireless thermistor in the abdominal cavity. Temperature in all phases was recorded every 5 minutes for 5 hours. Population body temperature (PBT) was recorded for approximately 8 days. Baseline body temperature is the final 24 hours of the PBT. All injectable drugs were given intramuscularly. The cats were administered drugs in four phases: 1) hydromorphone (H) 0.05, 0.1, or 0.2 mg kg(-1); 2) morphine (M) (0.5 mg kg(-1)), buprenorphine (BUP) (0.02 mg kg(-1)), or butorphanol (BUT) (0.2 mg kg(-1)); 3) ketamine (K) (5 mg kg(-1)) or ketamine (5 mg kg(-1)) plus hydromorphone (0.1 mg kg(-1)) (KH); 4) isoflurane in oxygen for 1 hour. Fifteen minutes prior to inhalant anesthetic, cats received either no premed (I), hydromorphone (0.1 mg kg(-1)) (IH), or hydromorphone (0.1 mg kg(-1)) plus ketamine (5 mg kg(-1)) (IHK). RESULTS: Mean PBT for all unmedicated cats was 38.9 +/- 0.6 degrees C (102.0 +/- 1 degrees F). The temperature of cats administered all doses of hydromorphone increased from baseline (p < 0.03) All four opioids (H, M, BUP and BUT) studied increased body temperature compared with baseline (p < 0.005). A significant difference was observed between baseline temperature values and those in treatment KH (p < 0.03). Following recovery from anesthesia, temperature in treatments IH and IHK was different from baseline (p < 0.002). CONCLUSIONS AND CLINICAL RELEVANCE: All of the opioids tested, alone or in combination with ketamine or isoflurane, caused an increase in body temperature. The increase seen was mild to moderate (<40.1 degrees C (104.2 degrees F) and self limiting. PMID- 20017818 TI - Comparison of a method using the HemoCue near patient testing device with a standard method of haemorrhage estimation in dogs undergoing spinal surgery. AB - OBJECTIVE: To compare an estimate of blood loss obtained using measurements from the Hemocue photometer with a standard estimate in dogs undergoing spinal surgery. STUDY DESIGN: Prospective clinical study. ANIMALS: Twenty-nine client owned dogs. METHODS: During surgery, blood and all lavage fluids were collected in the suction container and on to swabs. To prevent blood clot formation in the suction container, 10 mL citrate-phosphate dextrose adenine (CPDA) was added. At the end of the procedure, the total volume in the suction container was recorded. It was shaken to ensure uniformity and a 5 mL sample tested with the HemoCue photometer. Blood loss in the suction container was calculated as follows: Blood in suction (mL) = volume in bottle (mL) x [suction haemoglobin (Hb) concentration (g dL(-1))/pre-operative Hb concentration (g dL(-1))]. This volume was added to the estimated volume of blood on the swabs (weight of soaked swabs minus that of dry swabs) to provide the Hemocue estimate of total blood loss. A standard haemorrhage estimate was performed using the volume of fluid in the suction container at the end of surgery in excess of the total volume of lavage fluid available, minus 10 mL CPDA. This volume was added to the estimated volume of blood on the swabs to provide the standard estimate of total blood loss. Data were analyzed with a paired t-test. Retrospective power calculations demonstrated an 80% power to detect a mean difference of 25 mL between the two methods with a level of significance of 0.05. RESULTS: There was no significant difference in calculated blood loss between the two methods (p = 0.8, mean difference: -2 mL, 95% CI: -20 to 16 mL). CONCLUSIONS AND CLINICAL RELEVANCE: The HemoCue may be used to help estimate blood loss in dogs undergoing spinal surgery. PMID- 20017819 TI - Clinical pharmacology of methadone in dogs. AB - OBJECTIVE: To investigate the pharmacokinetics and effects of methadone on behaviour and plasma concentrations of cortisol and vasopressin in healthy dogs. STUDY DESIGN: Randomized, cross-over, experimental trial. ANIMALS: Nine adult dogs (beagle and beagle cross breeds), four males and five females. METHODS: Methadone hydrochloride, 0.4 mg kg(-1), was administered intravenously (IV) and subcutaneously (SC) with a crossover design. Drug and hormone analyses in plasma were performed using Liquid Chromatography-Electrospray Ionization-Tandem Mass Spectrometry and radioimmunoassay respectively. Behavioural data were collected using a standardized protocol. RESULTS: After IV administration, the plasma concentration of methadone at 10 minutes was 82.1 +/- 9.2 ng mL(-1) (mean +/- SD), the terminal half-life was 3.9 +/- 1.0 hours, the volume of distribution 9.2 +/- 3.3 L kg(-1) and plasma clearance 27.9 +/- 7.6 mL minute(-1) kg(-1). After SC administration, time to maximal plasma concentration was 1.26 +/- 1.04 hours and maximal plasma concentration of methadone was 23.9 +/- 14.4 ng mL(-1), the terminal half-life was 10.7 +/- 4.3 hours and bioavailability was 79 +/- 22%. Concentrations of both cortisol and vasopressin were increased for an hour following IV methadone. The observed behavioural effects of methadone were decreased licking and swallowing and an increase in whining after SC administration. The latter finding is notable as it can be misinterpreted as pain when methadone is used as an analgesic. CONCLUSION AND CLINICAL RELEVANCE: When methadone was administered by the SC route, the half-life was longer, but the individual variation in plasma concentrations was greater compared with IV administration. Increased frequency of whining occurred after administration of methadone and may be a drug effect and not a sign of pain. Cortisol and vasopressin concentrations in plasma may not be suitable for evaluating analgesia after methadone treatment. PMID- 20017820 TI - The use of an acoustic device to identify the extradural space in standing horses. AB - OBJECTIVE: To determine the usefulness of an acoustic device to confirm correct placement of extradural needles in horses. STUDY DESIGN: Prospective experimental study. ANIMALS: Twelve adult healthy horses weighing between 434 and 640 kg. MATERIALS AND METHODS: Horses were sedated, placed in stocks and subjected to caudal extradural anaesthesia using lidocaine 2%. For extradural puncture an 18 gauge Tuohy needle connected to an acoustic device via an electronic pressure transducer was used. This device allowed recording of the extradural pressure profile and the transformation of pressure changes into an audible signal. Extradural needle advancement was stopped and the local anaesthetic administered when either a sharp decrease in sound pitch occurred or a 'pop' sensation was felt. Correct needle placement was assessed clinically by the degree of extradural anaesthesia present using needle prick stimuli, loss of tail tone and occurrence of localized sweating. Descriptive statistical analysis was used for evaluation of the data. RESULTS: Extradural anaesthesia was successful in nine of 12 horses. In these horses during extradural needle advancement the pitch of the audible signal first increased and abruptly changed to a constant low pitched sound. In the three horses with unsuccessful punctures only a slight decrease of the pitch was noted. A 'pop' was clearly distinguished in five of nine horses with successful extradural puncture. In four horses a 'pop' was not present despite correct extradural needle placement. In three horses with incorrect needle placement a 'pop' was present. In the horses with correct needle positioning the median extradural pressure after the puncture was -0.8 kPa (-6 mmHg). CONCLUSION AND CLINICAL RELEVANCE: The acoustic device is useful to assist in the identification of successful or unsuccessful extradural needle placement in the horse. PMID- 20017821 TI - Bupivacaine 0.25% and methylene blue spread with epidural anesthesia in dog. AB - OBJECTIVE: To evaluate the extent sensory and motor blocks produced by the epidural injection of different volumes of 0.25% bupivacaine (Bu) with methylene blue (MB), in dogs. STUDY DESIGN: Prospective experimental trial. ANIMALS: Twenty healthy adult mongrel dogs, weighing 9.9 +/- 1.9 kg. METHODS: Dogs were randomly allocated into one of four groups that received 0.2, 0.4, 0.6 or 0.8 mL kg(-1) of an epidural solution containing 0.25% Bu and MB. Sensory block was evaluated against time by pinching the tail, hind limb interdigital web, toenail bases and the skin over the vertebral dermatomes. Motor block was assessed by ataxia, hind limb weight-bearing ability and by loss of muscle tone of the tail and pelvic limbs. Data were collected at 2, 5, 10, 15 and 30 minutes after the end of epidural injection. After the final time point, dogs were euthanatized and laminectomies were conducted to expose the extent of the dural dye staining. RESULTS: The volumes 0.2, 0.4, 0.6 and 0.8 mL kg(-1) of 0.25% Bu and MB blocked a mean of 5, 14.2, 20.2 and 21 dermatomes, respectively. The extent of the sensory block increased up to a volume of 0.6 mL kg(-1). Motor block was longer-lasting and more intense than sensory block. Complete dyeing of the spinal cord with MB was achieved in some dogs at 0.4 mL kg(-1) and all dogs at 0.6 mL kg(-1). CONCLUSIONS: The volume of anesthetic injected into the epidural space plays an important role in the quality of the epidural anesthesia. At 0.25%, bupivacaine provided an efficient sensory block at 0.6 mL kg(-1). CLINICAL RELEVANCE: Relatively high volumes (0.6 mL kg(-1)) of 0.25%, BU and MB were needed to produce an effective sensory and motor block caudal to the umbilicus, but all spinal cord segments were reached by MB at this dose. PMID- 20017822 TI - Antinociceptive, sedative and cardiopulmonary effects of subarachnoid and epidural xylazine-lidocaine in xylazine-sedated calves. AB - OBJECTIVE: To evaluate the antinociceptive, sedative and cardiopulmonary effects of subarachnoid and epidural administration of xylazine-lidocaine in xylazine sedated calves. STUDY DESIGN: Prospective, crossover study. ANIMALS: Six clinically healthy Holstein calves. MATERIALS: The calves were allocated randomly to receive two treatments, subarachnoid or epidural xylazine (0.025 mg kg(-1)) lidocaine (0.1 mg kg(-1)) diluted to a total volume of 5 mL with physiological saline. Prior to either epidural or subarachnoid injection, sedation was induced in all calves by intravenous administration of 0.1 mg kg(-1) xylazine. The quality and duration of antinociception and sedation were monitored. Areas of the cranial abdomen, umbilicus, and caudal abdomen were evaluated for antinociception using pinprick tests with a scoring system of 0-3 (0, none; 1, mild; 2, moderate; 3, complete). Sedation was assessed by using a 4-point scale (0, none; 1, mild; 2, moderate; 3, deep). The following cardiopulmonary variables were monitored: heart rate (HR), respiratory rate (f(R)), mean arterial pressure (MAP), blood pH, arterial partial pressure of oxygen (PaO(2)), partial pressure of carbon dioxide (PaCO(2)), bicarbonate (HCO(3)), base excess (BE), and oxygen saturation (SaO(2)). RESULTS: Xylazine sedation and subarachnoid xylazine-lidocaine resulted in significantly higher nociceptive block than the epidural technique. Moreover, subarachnoid xylazine-lidocaine induced a significantly longer duration of complete antinociception (median [IQR]) in the cranial abdomen (15.0 [15.0-30.0] versus 7.5 [1.3-10.0] minutes; p < 0.05) and umbilicus (45.0 [32.5-57.5] versus 10.0 [6.3-17.5] minutes; p < 0.05) compared with epidural xylazine-lidocaine. There was moderate sedation with both techniques. In both treatments, blood pH, MAP and PaO(2) decreased significantly, and PaCO(2) increased significantly during anaesthesia. No change was evident in HR, f(R), HCO(3,) BE, or SaO(2). CONCLUSION AND CLINICAL RELEVANCE: The subarachnoid injection provided better quality and longer duration of antinociception than epidural administration of the same doses of xylazine-lidocaine in xylazine-sedated calves, while cardiopulmonary depressant effects were observed with both regimens. PMID- 20017823 TI - Identification of single nucleotide polymorphisms within exon 1 of the canine mu opioid receptor gene. AB - OBJECTIVE: To determine the presence and frequency of single nucleotide polymorphisms (SNPs) within exon 1 of the canine mu-opioid receptor (MOR) gene. STUDY DESIGN: Prospective genetic analysis. ANIMALS: Seventy-five dogs of various breeds. METHODS: DNA was isolated from dog blood. Polymerase chain reaction (PCR) was performed to amplify exon 1 of the canine MOR gene using primers derived from a published sequence. PCR products of anticipated size were identified by gel electrophoresis, isolated and sequenced. RESULTS: Two SNPs were found within the examined region. One is 15 base pairs (bp) upstream (C-15A) of the protein-coding portion of the gene. The second is at position 207 (C207T); a synonymous mutation predicting unaltered protein sequence. The overall prevalence of the C-15A SNP was 43% (64/150 alleles). The overall prevalence of the C207T SNP was 26% (39/150 alleles). CONCLUSIONS AND CLINICAL RELEVANCE: Absence of haplotypes containing both an adenosine at position -15 and a thymine at position 207 suggests that these polymorphisms occurred independently from each other. How these SNPs influence variations in responses seen after opioid administration to dogs remain to be determined, however, our data indicates the C-15A SNP may play a role in opioid dysphoria. PMID- 20017824 TI - Fatal post-anaesthetic pneumothorax in a dog. PMID- 20017827 TI - Cleveland Clinic workshop on innovation in treatment of uremia: an introduction to the proceedings. PMID- 20017828 TI - More of the same: improving outcomes through intensive hemodialysis. AB - The typical dialysis patient faces both a poor quality of life and a significantly shortened survival. This is often blamed on "uremia." However, defining the clinical entity of uremia is surprisingly difficult. It represents the clinical sequelae of the effects of retention products, other effects of renal disease, and the effects of other comorbid conditions. The list of retention products that could act as uremic toxins is lengthy, but it would appear that urea itself does not contribute significantly to the uremic state. Larger molecular weight substances are likely the major contributors to the uremic milieu. Regardless of the causes, the uremic state persists in many patients who are reaching their dialysis adequacy targets as defined by urea clearance. This raises the possibility that more intensive hemodialysis could improve patient outcomes. Hemodialysis can be intensified by increasing dialysis efficiency without changing duration or frequency. Alternatively, hemodialysis duration, frequency, or both can be increased. All intensification methods increase small solute removal, but the removal of larger molecular weight retention products depends more upon treatment time. Modalities such as short daily hemodialysis, long intermittent hemodialysis, and quotidian nocturnal hemodialysis have been associated with a variety of clinical improvements, as well as improvements in quality of life and a lower standardized mortality ratio. However, the HEMO study approach of intensifying small solute clearance without significant modifications of the dialysis schedule does not appear to be effective. Future research will help to define the optimal treatment duration and frequency in hemodialysis patients. PMID- 20017829 TI - Renal cell therapy and beyond. AB - Although current dialysis techniques have transformed acute and chronic renal failure from uniformly fatal clinical disorders into treatable diseases, these therapies replace only the water and solute clearance function of the kidney and have reached a point where little further therapeutic improvement can be anticipated. In addition to their metabolic and endocrine functions, renal tubule cells presumably play an important role in the systemic inflammatory balance by participating in the complex and dynamic network of leukocyte action and pro- and anti-inflammatory cytokines. Loss of this function may result in a propensity to develop systemic inflammatory response syndrome (SIRS), multiorgan dysfunction, and a high risk of death in acute kidney injury (AKI), and may relate to chronic inflammatory state in end-stage renal disease (ESRD). A renal tubule cell assist device (RAD) containing animal or human renal tubule cells has been recently developed with the purpose of integrating the functions of tubule cells with the filtration function of current dialysis to offer a more complete renal replacement therapy. The viability and functionality of this device were confirmed in in vitro experiments and large animal studies, and recently the RAD's clinical therapeutic benefit was demonstrated with a series of FDA-approved human trials. Another novel synthetic membrane extracorporeal device that binds and inhibits circulating leukocytes has been developed with the purpose of reducing microvascular damage promoted primarily via activated circulating leukocytes in AKI and SIRS. This device, called a selective cytopheretic inhibitory device, mimics immunomodulation and duplicates RAD efficiency in preliminary studies. Both devices may become comprehensive treatments, replacing full renal function and correcting inflammatory imbalance in patients with acute and chronic renal disorders. PMID- 20017830 TI - Convective therapies for removal of middle molecular weight uremic toxins in end stage renal disease: a review of the evidence. AB - The increasing number of patients requiring renal replacement therapy poses a challenge to maintain quality of care in the setting of limited resources. The commonly used modalities include hemodialysis and peritoneal dialysis, and using a "urea-centric" model to increase the clearance of small molecular weight uremic toxins beyond current guidelines does not appear to confer additional clinical benefits in terms of morbidity and mortality. Convective therapies including hemofiltration and hemodiafiltration, which also have higher middle molecule clearances, might offer significant benefits compared to diffusive therapy. We review the available evidence on convective therapies and their effects on middle molecular weight uremic toxins, particularly beta2 microglobulin because of its known associated morbidity. It is the authors' opinion that more emphasis should be placed on true uremic toxins such as beta2 microglobulin rather than surrogate uremic toxins like urea. Larger studies are also needed to study the merits of convective therapies, with a focus on cardiovascular morbidity and mortality. PMID- 20017831 TI - Sorbents in treatment of uremia: a short history and a great future. AB - To regenerate dialysate in hemodialysis for kidney failure, it is necessary to remove all uremic toxins, avoid excessive removal of various electrolytes, and supply beneficial components such as bicarbonate. Surprisingly, a simple collection of components can accomplish this task very well: charcoal, urease, cation exchanger, and anion exchanger. From the early days of use of the Redy system and the Sorb cartridge, improvements have been made both in machine design and column chemical function. This article describes the chemical function of the Sorb cartridge, recent improvements, and the methods to predict the chemical function of the column. It also describes a number of improvements made in dialysis machines using sorbent regeneration of the dialysate. Some ongoing improvements in sorbent chemistry are also described. PMID- 20017832 TI - Blood-membrane interactions during dialysis. AB - In extracorporeal renal replacement therapies, the dialyzer is not only the site at which solute removal occurs but also the extracorporeal circuit component having the largest surface area exposed to blood. Therefore, it is not surprising that interactions between blood components and the dialyzer membrane influence the dialysis procedure in several ways. Based on engineering principles, fluid flow along a surface such as membrane results in the development of a boundary layer which can influence solute removal. Furthermore, the exposure of blood to any extracorporeal artificial surface results in the activation of several pathways within the body, including those involving coagulation and complement activation. One of the byproducts of this generalized activation process is protein adsorption to the membrane surface, another phenomenon which can have a significant impact on solute removal. In this article, a detailed review of the ways in which blood-membrane interactions influence solute removal during hemodialysis and related therapies is provided. The influences of secondary membrane formation and boundary layer/concentration polarization effects on solute removal are specifically discussed. Furthermore, the importance of adsorption as a specific removal mechanism for low-molecular weight proteins by highly permeable synthetic membranes is highlighted. PMID- 20017833 TI - Cellular therapy of kidney diseases. AB - The understanding of cellular sources of kidney regeneration has rapidly evolved in the last decade. It is now believed that regeneration occurs predominantly from cells that reside within the injured kidney, with minimal contribution from extra-renal cells. We now know that improved kidney regeneration seen following exogenous administration of stem cells occur predominantly by noncellular paracrine mechanisms. Of all extra-renal stem cells, mesenchymal stem cells (MSC) are the most promising stem cell type for treating kidney diseases. There is an ongoing clinical trial evaluating safety and efficacy of MSC in treating acute kidney injury (AKI). Results of this trial are expected to bring use of MSC closer to the clinical realm. An improved understanding of the small molecules that facilitate kidney regeneration and are secreted by MSC will likely result in the development of new therapies for treating AKI. Identification of adult stem cell markers will result in improved understanding of pathophysiology of kidney diseases and could lead to the development of new cellular therapies. Directed differentiation of stem cells into desired cell types such as erythropoietin producing cells will allow selective replacement of lost kidney function. Cell based therapies for patients with chronic kidney disease are presently in proof of-principle stage and are expected to evolve in the coming years with improved understanding of stem cell biology. Technological advancement in cellular therapy is expected to provide improved therapeutic options for patients with kidney diseases in the near future. PMID- 20017834 TI - Uremic toxicity, oxidative stress, and hemodialysis as renal replacement therapy. AB - Patients with uremia are subject to greatly increased cardiovascular risk that cannot be completely explained by traditional cardiovascular risk factors. An increase in oxidative stress and inflammation has been proposed as contributory nontraditional uremic cardiovascular risk factors. Oxidative stress reflects the balance between oxidant generation and antioxidant defense mechanisms. Reduction/oxidation (redox) reactions may result in a stochastic process leading to oxidation of neighboring macromolecules. However, in many instances the reactive oxygen species target particular amino acid residues or lipid moieties. This provides a mechanism by which increased oxidative stress and/or alteration of antioxidant mechanisms can alter cell signaling. In individuals with advanced chronic kidney disease, the redox balance is not in equilibrium and is tipped toward oxidation resulting in the dysregulation of cellular process with subsequent vascular and tissue injury. In this review, the major oxidant and antioxidant pathways and the biomarkers to assess redox status in uremia are discussed, as well as the data linking the pathogenesis of oxidative stress, inflammation, cardiovascular events, and the progressive loss of kidney function in chronic kidney disease. PMID- 20017836 TI - Inflammation and insulin resistance as novel mechanisms of wasting in chronic dialysis patients. AB - Over the last decade, there have been no proven therapies to lower the mortality and morbidity risk for chronic dialysis patients. One of the most important determinants of this poor clinical outcome is protein energy wasting (PEW), a unique and highly prevalent nutritional and metabolic abnormality primarily characterized by increased protein breakdown in the skeletal muscle compartment. Although the etiology and mechanisms leading to increased protein breakdown in chronic dialysis patients are complex and mostly ill-defined, two well-recognized and presumably interrelated metabolic abnormalities, insulin resistance and chronic inflammation, are likely to play a critical role in the pathogenesis of this condition. Multiple studies demonstrate the anabolic effects of insulin that extend beyond simple carbohydrate metabolism. Insulin is a mediator of accelerated protein breakdown in the catabolic condition such as advanced kidney disease. Chronic inflammation, a condition known to cause muscle catabolism in experimental conditions, has a strong association with advanced kidney disease in epidemiologic studies. Chronic inflammation is also known to induce insulin resistance, primarily by the induction of proinflammatory cytokines. The protein catabolic effects of inflammation and insulin resistance involve common cellular pathways. Thus, it is reasonable to speculate that chronic inflammation of advanced kidney disease mediates its protein catabolic effects by inducing insulin resistance of protein metabolism at both the physiologic and cellular levels. Modulating inflammatory response or insulin signaling by pharmacologic interventions could allow us to clarify the mechanisms contributing to the development of PEW in the setting of these particular metabolic derangements. PMID- 20017837 TI - Technical approaches toward ambulatory ESRD therapy. AB - Current efforts to prolong dialysis and make it ambulatory may, but need not, follow the established countercurrent contact of blood and dialysate through a membrane. Avoiding anticoagulation and addressing decremented performance of membranes used over long times suggests gentler blood contact and greatly reduced contact areas. This paper describes a microfluidic fluid-to-fluid contact system, still under development, and suggests that initial attempts at ambulatory support of end-stage renal disease patients may be limited to interdialytic volume control, perhaps with a reduced frequency of dialysis which would then be used only to remove accumulated solutes. PMID- 20017835 TI - Causes of dysregulation of lipid metabolism in chronic renal failure. AB - End-stage renal disease (ESRD) is associated with accelerated atherosclerosis and premature death from cardiovascular disease. These events are driven by oxidative stress inflammation and lipid disorders. ESRD-induced lipid abnormalities primarily stem from dysregulation of high-density lipoprotein (HDL), triglyceride rich lipoprotein metabolism, and oxidative modification of lipoproteins. In this context, production and plasma concentration of Apo-I and Apo-II are reduced, HDL maturation is impaired, HDL composition is altered, HDL antioxidant and anti inflammatory functions are depressed, clearance of triglyceride-rich lipoproteins and their atherogenic remnants is impaired, their composition is altered, and their plasma concentration is elevated in ESRD. The associated defect in HDL maturation is largely caused by acquired lecithin-cholesterol acyltransferase deficiency while its triglyceride enrichment is due to hepatic lipase deficiency. Hypertriglyceridemia, abnormal composition, and impaired clearance of triglyceride-rich lipoproteins and their remnants are mediated by down-regulation of lipoprotein lipase, hepatic lipase, very low-density lipoprotein (VLDL) receptor, and LDL receptor-related protein, relative reduction in ApoC-II/ApoC III ratio, up-regulation of acyl-CoA cholesterol acyltransferase, and elevated plasma level of cholesterol ester-poor prebeta HDL. Impaired clearance and accumulation of oxidation-prone VLDL and chylomicron remnants and abnormal LDL composition in the face of oxidative stress and inflammation favors their uptake by macrophages and resident cells in the artery wall. The effect of heightened influx of lipids is compounded by impaired HDL-mediated reverse cholesterol transport leading to foam cell formation which is the central event in atherosclerosis plaque formation and subsequent plaque rupture, thrombosis, and tissue damage. PMID- 20017838 TI - Bottom-up nanotechnology: the human nephron filter. AB - Over one million patients worldwide have end-stage renal disease and require dialysis or kidney transplantation. Despite the availability of these forms of renal replacement therapy for nearly four decades, mortality and morbidity are high and patients often have a poor quality of life. We have developed a human nephron filter (HNF) utilizing bottom-up nanotechnology that would eventually make feasible a continuously functioning, wearable or implantable artificial kidney. The device consists of two membranes operating in series within one device cartridge. The first membrane mimics the function of the glomerulus, using convective transport to generate a plasma ultra filtrate, which contains all solutes approaching the molecular weight of albumin. The second membrane mimics the function of the renal tubules, selectively reclaiming designated solutes to maintain body homeostasis. No dialysis solution is used in this device. The HNF has been computer modeled, and operating 12 hour per day, 7 days per week the HNF provides the equivalent of 30 ml/min glomerular filtration rate (compared to half that amount for conventional thrice-weekly hemodialysis). The HNF system, by eliminating dialysate and utilizing a novel membrane system created through applied nanotechnology, represents a breakthrough in renal replacement therapy based on the functioning of native kidneys. The enhanced solute removal and wearable design should substantially improve patient outcomes and quality of life. PMID- 20017839 TI - The implantable artificial kidney. AB - The confluence of an increasing prevalence of end-stage renal disease (ESRD), clinical trial data suggestive of benefit from quotidian dialysis, and ongoing cost/benefit reanalysis of healthcare spending have stimulated interest in technological improvements in provision of ESRD care. For the last decade, our group has focused on enabling technologies that would permit a paradigm shift in dialysis care similar to that brought by implantable defibrillators to arrhythmia management. Two significant barriers to wearable or implantable dialysis persist: package size of the dialyzer and water requirements for preparation of dialysate. Decades of independent research into highly efficient membranes and cell-based bioreactors culminated in a team effort to develop an implantable version of the University of Michigan Renal Assist Device. In this review, the rationale for the design of the implantable artificial kidney is described. PMID- 20017840 TI - A novel technique for tethered dialysis catheter removal using the laser sheath. AB - Traction and cutdown techniques can successfully remove a tunneled dialysis catheter (TDC) in a great majority of patients. However, these methods may not be successful in patients with catheters that are tethered or attached to the central veins or the atrium. A forceful application of traction can lead to catheter breakage with subsequent retention of the broken piece and carries a potential risk of vascular and atrial wall avulsion. Open thoracotomy has been employed to remove an attached TDC. However, this procedure is invasive and bears a significant morbidity. This report presents three cases of tethered TDCs that underwent laser sheath extraction. The TDCs had been in place for an average of 26 months. The patients underwent initial unsuccessful removal attempt using the traction method with surgical exploration all the way to the venotomy site. The laser technique that is used to remove pacemaker/implantable cardioverter defibrillator leads was then applied to these stuck catheters. All three catheters were successfully removed without any damage to the catheter, central veins, or the right atrium. There were no retained catheter fragments left in the central veins or the atrium. One patient demonstrated a significant thrombus that extended from the tip of the catheter all the way to the right ventricle. The external sheath of the laser device successfully aspirated the thrombus. There were no procedure-related complications. In this small series, a laser sheath successfully extracted tethered dialysis catheters. The study found the procedure to be effective, easy to perform, and minimally invasive. We suggest that this approach be considered for the removal of tethered catheters that cannot be removed using traditional approaches. PMID- 20017841 TI - Focused vascular access education to reduce the use of chronic tunneled hemodialysis catheters: results of a network quality improvement initiative. AB - Tunneled hemodialysis catheters (TDCs) carry the highest mortality risk for chronic hemodialysis patients of any access modality. Recent data have emphasized that mortality risk decreases when these devices are discontinued. Herein, we present the results of a gap-reduction assisted catheter elimination strategy that Network 7 employed as its quality improvement initiative to reduce the use of TDCs. Hemodialysis facilities with high catheter rates (>90 days) were identified. Interventions included focused vascular access education, monthly follow-up and site visits to assist the facility catheter reduction program. The "goal" of interventions was defined as the gap-reduction of 50% from the baseline catheter rate to the Network mean plus sustainability of catheter reduction for at least 3 consecutive months. Fifteen facilities (n = 891) were identified with high catheter rates (31.5 +/- 5.3%) in May 2006. Interventions resulted in a catheter reduction to 12.2 +/- 8.5% in May 2007 (p = 0.0001). Five of the 15 facilities (n = 280) achieved the goal (preintervention = 31.7 +/- 5.3%, postintervention = 8.7 +/- 2.8%, p = 0.001). In May 2007, eight additional facilities (n = 438) with high catheter rates (31.7 +/- 7.8%) were added to the 10 that failed to achieve the goal previously. Interventions employed in these 18 facilities (n = 1,049) resulted in catheter reduction in all (preintervention = 31.5 +/- 5.5%, postintervention = 16.2 +/- 5%, p = 0.01). Five of these 18 met the goal (preintervention = 32 +/- 8%, postintervention = 5.9 +/- 4.3%). Overall, all 23 facilities (n = 1,329) demonstrated catheter reduction postintervention (preintervention = 31.6 +/- 6%, postintervention = 13.9 +/- 6%, p = 0.001), and 10/23 (43%) met the project goal (preintervention = 31.9 +/- 6%, postintervention = 7.3 +/- 4%, p = 0.002). Medical director's involvement had a positive impact in achieving the goal (p = 0.003). The presence or absence of a vascular access coordinator did not affect catheter reduction. The results of this analysis reveals that an organized approach implemented by an ESRD Network can have a significant impact in reducing catheter use. PMID- 20017842 TI - Retrograde angiography and the risk of arteriovenous fistula perforation. AB - Assessment of the inflow segment of an arteriovenous fistula is mandatory for optimum access evaluation. Retrograde angiography (RA) is routinely used to image this region of the access system. Traditionally, RA is performed by manually occluding the outflow track of an arteriovenous fistula (AVF) and forcing the radiocontrast into the inflow segment against arterial pressure. While this technique is largely successful in visualizing the juxta-anastomotic region, anastomosis and a portion of the feeding artery, the approach carries a potential risk of vascular rupture between the occluded portion of the fistula and the anastomosis. This article presents six cases of fistulas that suffered vascular rupture during RA. In three cases, vascular damage occurred prior to the application of angioplasty. The remaining cases suffered perforation after angioplasty. Balloon tamponade was successful in salvaging two fistulas. Another AVF with a perforation did not require any intervention to maintain flow. The complication was successfully managed in one AVF by the insertion of an endovascular stent graft. Two fistulas were lost due to vascular damage. This report demonstrates that RA performed by occluding the outflow track of an AVF to assess the results of angioplasty of an inflow stenosis can result in vascular rupture. There is a risk of such adversity even before the application of angioplasty in this region. It is suggested that the first of these be avoided and the latter be done with care. PMID- 20017843 TI - On the retrograde occlusive arteriogram. PMID- 20017844 TI - Risk of bacterial contamination from citrate catheter locks. PMID- 20017845 TI - Briefly noted: Renal transplantation. PMID- 20017846 TI - Tissue engineering in wound repair: the three "R"s--repair, replace, regenerate. AB - Horses are predisposed to traumatic wounds that can be labor intensive and expensive to manage. Skin has a considerable potential for efficient and functional repair however, while cutaneous repair is a regenerative process in the fetus, this capability declines in late gestation as inflammation and scarring alter the outcome of healing. The historical gold standard for replacement of lost skin is the autologous skin graft. However, the horse's lack of redundant donor skin limits the practicality of full-thickness grafting to smaller wounds; moreover, graft failure is relatively common in equine patients as a result of infection, inflammation, fluid accumulation beneath the graft, and motion. Tissue engineering has emerged as an interdisciplinary field with the aim to regenerate new biological material for replacing diseased or damaged tissues or organs. In the case of skin, the ultimate goal is to rapidly create a construct that effects the complete regeneration of functional skin, including all its layers and appendages. Moreover, an operational vascular and nervous network, with scar-free integration within the surrounding host tissue, is desirable. For this to be achieved, not only is an appropriate source of cells required, but also a scaffold designed from natural or synthetic polymers. The newly created tissue might finally meet the numerous needs and expectations of practitioners and surgeons managing a catastrophic wound in a horse. PMID- 20017847 TI - Biologic behavior and clinical outcome of 25 dogs with canine appendicular chondrosarcoma treated by amputation: a Veterinary Society of Surgical Oncology retrospective study. AB - OBJECTIVE: To characterize biologic behavior, clinical outcome, and effect of histologic grade on prognosis for dogs with appendicular chondrosarcoma treated by amputation alone. STUDY DESIGN: Case series. ANIMALS: Dogs (n=25) with appendicular chondrosarcoma. METHODS: Medical records were searched to identify dogs with appendicular chondrosarcoma treated by limb amputation alone. Information recorded included signalment, anatomic location, radiographic appearance, and development of metastasis. Histopathologic diagnosis was confirmed and graded (1, 2, or 3). Survival curves were generated by the Kaplan Meier method and the association between covariates (gender, age, weight, and tumor grade) and survival were evaluated using the univariate proportional hazards model. RESULTS: Histopathology slides were available for 25 dogs. Rates of pulmonary metastasis were as follows: grade 1-0%, grade 2-31%, and grade 3 50%. Overall median survival time (MST) was 979 days. Age, weight, and sex were not significantly associated with survival (P=.16; .33; and .31, respectively). Survival was significantly associated with tumor grade (P=.008), with dogs with tumor grade of 1, 2, and 3 having MSTs of 6, 2.7, and 0.9 years, respectively. CONCLUSION: Canine appendicular chondrosarcoma can be treated effectively with amputation alone. Low to intermediate grade chondrosarcoma has a good prognosis, whereas high-grade tumors appear to behave aggressively. CLINICAL RELEVANCE: The overall prognosis for appendicular chondrosarcoma is better than that of appendicular osteosarcoma treated by amputation alone or in combination with chemotherapy. PMID- 20017848 TI - Interobserver repeatability of measurements on computed tomography images of lax canine hip joints from youth to maturity. AB - OBJECTIVES: To determine interobserver repeatability of measurements on computed tomography (CT) images of lax canine hip joints at different ages and in the presence of degenerative joint disease at maturity. STUDY DESIGN: Longitudinal observational investigation. ANIMALS: Sibling crossbreed hounds. METHODS: Pelvic CT was performed at 20, 24, 32, 48, 68, and 104 weeks of age. Measures were performed on 3 contiguous two-dimensional (2D) transverse CT images of both hips at each time point by 3 investigators. Center-edge angle (CEA), horizontal toit externe angle (HTEA), ventral (VASA), dorsal (DASA), and horizontal (HASA) acetabular sector angles, acetabular index (AI), and percent femoral head coverage (CPC) were measured. Interobserver repeatability was quantified with the intraclass correlation coefficient (ICC). Satisfactory repeatability was considered when ICC >or=0.75. RESULTS: DASA, CEA, and CPC were repeatable in all age groups. HASA and HTEA were repeatable for all but 1 time point. At 20 weeks of age, all measures but AI were repeatable, and at 104 weeks of age, DASA, CEA, CPC, and HASA were repeatable. Measures were repeatable in hips with and without degenerative changes with the exceptions of AI and HASA in normal hips and VASA and HTEA in osteoarthritic hips. CONCLUSIONS: Most 2D CT measurements examined were repeatable regardless of age or joint disease. CLINICAL RELEVANCE: Two dimensional CT measures may augment current techniques for assessing joint changes in lax canine hips. PMID- 20017849 TI - Use of propofol-xylazine and the Anderson Sling Suspension System for recovery of horses from desflurane anesthesia. AB - OBJECTIVE: To characterize the behavior of horses recovering in the Anderson Sling Suspension System after 4 hours of desflurane anesthesia and postdesflurane intravenous (IV) administration of propofol and xylazine. STUDY DESIGN: Experimental study. ANIMALS: Healthy horses (n=6), mean+/-SEM age 12.3+/-1.8 years; mean weight 556+/-27 kg. METHODS: Each horse was anesthetized with xylazine, diazepam, and ketamine IV and anesthesia was maintained with desflurane in O(2). At the end of 4 hours of desflurane, each horse was positioned in the sling suspension system and administered propofol-xylazine IV. Recovery events were quantitatively and qualitatively assessed. Venous blood was obtained before and after anesthesia for biochemical and propofol analyses. RESULTS: Anesthetic induction and maintenance were without incident. Apnea commonly accompanied propofol administration. All horses had consistent recovery behavior characterized by a smooth, careful, atraumatic return to a standing posture. CONCLUSIONS: Results of this study support careful, selective clinical use of desflurane, propofol-xylazine, and the Anderson Sling Suspension System to atraumatically transition horses with high anesthetic recovery risk to a wakeful standing posture. CLINICAL RELEVANCE: Technique choices to facilitate individualized, atraumatic recovery of horses from general anesthesia are desirable. Use of IV propofol and xylazine to transition horses from desflurane anesthesia during sling recovery to standing posture may facilitate improved recovery management of high-injury risk equine patients requiring general anesthesia. PMID- 20017850 TI - Postoperative race performance is not correlated with degree of surgical abduction after laryngoplasty in National Hunt Thoroughbred racehorses. AB - OBJECTIVES: To (1) assess the degree of arytenoid cartilage abduction lost after laryngoplasty (LP) in Thoroughbred National Hunt racehorses and (2) to correlate postoperative racing performance with degree of arytenoid abduction after LP. STUDY DESIGN: Case series. ANIMALS: National Hunt Thoroughbred racehorses (n=68). METHODS: Grade of postoperative arytenoid abduction for National Hunt racehorses that had LP with ventriculocordectomy was assessed at 1 day, 6 days, and 6 weeks after LP. Race records were analyzed to ascertain if there was correlation between the degree of arytenoid cartilage abduction and various measures of race performance (return to racing postoperatively, total earnings in 5 races immediately postoperatively, and lifetime number of starts postoperatively). RESULTS: Median postoperative arytenoid abduction was grade 2 on day 1 but had decreased to grade 3 by 6 weeks. Horses with grades 1, 2, and 3 abduction 1 day after surgery had median losses of 1, 1, and 0.5 abduction grades, respectively, at 6 weeks. Horses with grade 1 abduction on day 1 were significantly more likely to lose abduction by day 6 after surgery than horses with grade 3 abduction on day 1. There was no statistically significant correlation between the postoperative grade of arytenoid abduction at any time point and earnings in 5 races after surgery, likelihood of racing postoperatively, or total number of lifetime race starts postoperatively. CONCLUSIONS: Horses with maximal (grade 1) surgical arytenoid abduction are significantly more likely to suffer postoperative loss of abduction than those with grade 3 abduction. Postoperative grade of abduction does not appear significantly correlated with markers of racing performance in National Hunt racehorses; however, very few horses with poor (grade 4 or 5) abduction were included and thus conclusions regarding racing performance in such horses cannot be drawn from this study. CLINICAL RELEVANCE: Seemingly, most horses with grade 3 laryngeal abduction can race successfully and perhaps surgeons should not be disillusioned by the appearance of only moderate (grade 3) abduction in the long term after LP in racehorses. PMID- 20017851 TI - Race performance after laryngoplasty and ventriculocordectomy in National Hunt racehorses. AB - OBJECTIVE: To evaluate the effect of laryngoplasty (LP) on race performance in longer distance (National Hunt) Thoroughbred racehorses. STUDY DESIGN: Case control study. ANIMALS: National Hunt Thoroughbred racehorses (n=71) and race matched controls (n=126). METHODS: Race records for National Hunt racehorses that had LP and ventriculocordectomy were analyzed and racing performance was compared with race-matched controls. RESULTS: Sixty-three of 71 cases were matched with controls. Postoperatively, 78% of cases raced, 47% improved their individual performance and cases were as likely to start in 1 or 3 races as controls. In the 5 preoperative races, case horses earned less prize money than race-matched controls but there was no difference in prize money earned in 5 postoperative races between groups. Cases started in significantly fewer total (lifetime) races both before and after the date of surgery compared with controls. CONCLUSIONS: LP seemingly restored short-term postoperative racing performance of National Hunt horses to a level comparable with that of a matched control population; however, the career "longevity" of case horses appears to be shorter than that of control horses. CLINICAL RELEVANCE: LP appears to be a suitable treatment for recurrent laryngeal neuropathy in National Hunt racehorses. PMID- 20017852 TI - Bilateral iliopsoas muscle contracture and spinous process impingement in a German Shepherd dog. AB - OBJECTIVE: To report diagnosis and treatment of bilateral iliopsoas muscle contracture in a dog with spinous process impingement. STUDY DESIGN: Case report. ANIMALS: German Shepherd dog. METHODS: A dog with chronic progressive lameness, flexion contracture of the coxofemoral joints, severe pain, and decreased femoral reflexes had severe spondylosis bridging the vertebral bodies from L1 to L4 and enlarged dorsal spinous processes from T8 to L6 with impingement and bony proliferation. Ultrasonographic and magnetic resonance imaging (MRI) findings were consistent with fibrosis, mineralization, and atrophy of the iliopsoas muscles bilaterally which was treated by staged tenectomy of the insertions of the iliopsoas muscles. RESULTS: Because of severe perivascular fibrosis, the femoral vessels required ligation. Bilateral iliopsoas muscle tenectomy improved gait and provided pain relief. Histologic findings were consistent with fibrotic myopathy. CONCLUSIONS: Slow progression of severe clinical signs observed bilaterally in this dog differs from previous reports of iliopsoas myopathy. Findings were similar to the fibrotic myopathy of the gracilis or semitendinosus muscles described in dogs. CLINICAL RELEVANCE: Iliopsoas muscle abnormalities should be considered in dogs with limited hip extension and pain. MRI is useful for diagnosing muscle fibrosis. Iliopsoas tenectomy may improve clinical function in dogs with fibrotic myopathy. PMID- 20017853 TI - Biomechanical analysis of stacked plating techniques to stabilize distal radial fractures in small dogs. AB - OBJECTIVE: To evaluate the fatigue life of partially stacked and fully stacked (1.5/2.0 and 2.0/2.7 mm) veterinary cuttable plates (VCP) in a fracture gap model of the distal aspect of the radius. STUDY DESIGN: In vitro biomechanical study. METHODS: Constructs (n=4/group) were assembled for each of 8 groups using 8-hole plates (1.5/2.0 and 2.0/2.7 mm VCP) in the following configurations: unstacked; 2 hole stacked centered over the gap (COG); 4-hole stacked COG; and fully stacked. Plate(s) were secured to 2 separate polyvinylchloride pipe lengths, mounted to a mechanical testing system with a custom jig, and were loaded in axial compression for 10(6) cycles at 10 Hz or until failure at 6-60 N for the 1.5/2.0 mm VCP and 10-100 N for the 2.0/2.7 mm VCP. Differences in number of cycles, stiffness, and failure mode were recorded. RESULTS: All construct failures occurred through a screw hole adjacent to the gap. Fully stacked and 4-hole stacked 1.5/2.0 and 2.0/2.7 mm VCP withstood 10(6) cycles. Fatigue life and stiffness of the 1.5/2.0 or 2.0/2.7 mm unstacked constructs were significantly less than the other constructs. Differences were identified in stiffness among the 1.5/2.0 mm stacked constructs and in fatigue life among the 2.0/2.7 mm VCP stacked constructs. CONCLUSION: Four-hole partially stacked VCP (either 1.5/2.0 or 2.0/2.7 mm) have comparable mechanical properties to fully stacked VCP. CLINICAL RELEVANCE: Partial stacking of 2 holes of VCP on both sides of the fracture gap may provide sufficient strength for healing, but this premise must be tested in vivo. PMID- 20017854 TI - Thoracoscopic subphrenic pericardectomy using double-lumen endobronchial intubation for alternating one-lung ventilation. AB - OBJECTIVE: To report the technique, complications, and effectiveness of thoracoscopic subphrenic pericardectomy (SPP) using double-lumen endobronchial intubation for alternating 1-lung (OLV) in healthy dogs. STUDY DESIGN: Prospective cohort study. ANIMALS: Mature purpose-bred dogs (n=7). METHODS: Bronchoscope-assisted placement of a left-sided double-lumen endobronchial tube, immediately before surgery, allowed intraoperative alternation of ventilation between lung fields. A camera portal was established in a subxyphoid location. Two instrument portals were established at the 4th-6th intercostal spaces on the right and left sides. A vessel-sealing device was used to create the subphrenic pericardectomy. After termination of the procedure, dogs were humanely euthanatized under anesthesia and necropsy performed. In each dog, the extent of pericardectomy and any complications were evaluated. RESULTS: Technical difficulties with tube placement occurred in 4 dogs, but alternating OLV was achieved in all dogs and SPP completed successfully. Median surgical time was 87.5 minutes (range, 80-105 minutes). At necropsy, 0.5-2 cm of pericardial tissue remained ventral to the intact phrenic nerve in 6 dogs; in 1 dog, the phrenic nerve was transected on the left side only. CONCLUSIONS: Thoracoscopic subphrenic pericardectomy is a technically feasible procedure in healthy dogs. Double-lumen endobronchial intubation allowed alternating OLV without intraoperative bronchoscopically guided tube manipulation in all but 1 dog. CLINICAL RELEVANCE: Thoracoscopic subphrenic pericardectomy could potentially be used for management of conditions where relief of pericardial constriction or access to intrapericardial structures is desired. PMID- 20017855 TI - A laparoscopic-sutured gastropexy technique in dogs: mechanical and functional evaluation. AB - OBJECTIVE: To describe a laparoscopic-sutured gastropexy technique in dogs and evaluate the tensile strength of the adhesion and effects on gastric function. STUDY DESIGN: Experimental study. ANIMALS: Female beagle dogs (n=7). METHODS: A laparoscopic-sutured gastropexy technique was evaluated by ex vivo tensile distraction tests 10 weeks after surgery. The effect of the adhesion on gastric emptying, mucosal permeability, and systemic inflammation were evaluated by monitoring the C-reactive protein (CRP) and sucrose permeability, and by radiographic evaluation of gastric emptying 2 weeks before and 10 weeks after surgery. RESULTS: Mean (+/-SD) tensile force to disrupt adhesions was 51.1+/-16.4 N. There was no significant postoperative increase in CRP concentration or change in sucrose permeability. The area under the curve representing the postprandial decrease in gastric radiographic area increased by 11% after gastropexy. CONCLUSIONS: This laparoscopic gastropexy technique had appropriate mechanical and functional characteristics with limited morbidity. CLINICAL RELEVANCE: This laparoscopic-sutured gastropexy provides adhesion strength comparable with other gastropexy techniques tested at 10 weeks postoperatively. Only minor changes in gastric emptying were observed 10 weeks after surgery. PMID- 20017856 TI - Evaluation of a transcondylar toggle system for stabilization of the cranial cruciate deficient stifle in small dogs and cats. AB - OBJECTIVE: To evaluate use of a transcondylar toggle system (TCTS) for stabilization of the cranial cruciate ligament (CrCL) deficient stifle in small dogs and cats. STUDY DESIGN: Prospective clinical study. ANIMALS: Small dogs (<7 kg; n=14) and cats (2) with CrCL-associated lameness of <3 months duration and a tibial plateau angle <32 degrees. METHODS: Affected animals had an extracapsular CrCL repair using the TCTS. Lameness score, muscle atrophy, osteoarthritis (OA) score, and range of motion (ROM) were evaluated preoperatively, and at 6 weeks and 7-10 months postoperatively. RESULTS: Operative time was 75 +/- 16 minutes. Fifty-six percent required >1 bone tunnel attempts. One dog required revision at 2 weeks because of suture loosening. All stifles were stable at 6 weeks postoperatively. Fifteen animals were available for follow-up (7-10 months). Lameness improved significantly at 6 weeks (P<.0001), whereas muscle atrophy was worse at 6 weeks (P=.008) but improved at 7-10 months (P<.0001). OA scores were unchanged at 6 weeks (P=.08) but were significantly worse at 7-10 months (P<.0001). ROM remained unchanged at 6 weeks (P=1) and 7-10 months (P=.6). CONCLUSIONS: The medially placed toggle provides a reliable short-term proximal anchor for the extracapsular suture with outcomes similar to other extracapsular techniques. The aiming device and drill bit are not recommended in their current form. CLINICAL RELEVANCE: The TCTS appears to be a well-tolerated technique for proximal suture anchoring in extracapsular CrCL repair in small dogs and cats where instrumentation and anatomic constraints preclude other techniques. PMID- 20017857 TI - Hypophosphatemia in cats after renal transplantation. AB - OBJECTIVE: To report the prevalence of hypophosphatemia after renal transplantation in a historical cohort of cats. DESIGN: Case series. ANIMALS: Cats (n=86) that received a renal allograft. METHODS: Medical records (January 200-June 2006) were reviewed. Signalment, clinical signs, pre- and postoperative diet, pre- and postoperative clinicopathologic variables, renal histopathology, and outcome were retrieved. Prevalence, onset, duration, treatment and associated clinical signs of hypophosphatemia were recorded. A chi(2) test was used to compare hemolysis frequency between cats with normal serum phosphorus concentration or a single spurious low serum phosphorus concentration for <24 hours duration (group 1) and confirmed hypophosphatemia for >24 hours (group 2). A Cox proportional hazards model was used to evaluate the effects of hypophosphatemia on survival while controlling for other potentially confounding variables (age, sex, weight, body condition score, and pre- and 24 hours postoperative clinicopathologic variables). RESULTS: Eighty-six cats (mean age, 7.7 years) were identified. Hypophosphatemia occurred in 32 cats (37%), with a median onset of 2 days and median duration of 4 days. Treatment was initiated in 48 (56%) of hypophosphatemic cats. Survival and hemolysis frequency was not significantly different between groups, and no risk factors were identified. CONCLUSION: Hypophosphatemia occurs in cats after renal transplantation and does not affect survival. CLINICAL RELEVANCE: The clinical importance of hypophosphatemia in renal transplant recipients remains unknown. PMID- 20017858 TI - Comparison of three drilling techniques for carpometacarpal joint arthrodesis in horses. AB - OBJECTIVE: To evaluate 3 drilling techniques for arthrodesis of the equine carpometacarpal (CMC) joint. STUDY DESIGN: Experimental study. SAMPLE POPULATION: Cadaveric equine forelimbs (n=15). METHODS: Limbs were divided into 3 groups (5 limbs each) to evaluate 3 drilling techniques: (1) use of a 4.5 mm drill bit inserted into the joint through 4 entry points and moved in a fanning motion; (2) a 5.5 mm drill bit inserted through 2 entry points to create 3 nonfanned drill tracts (3 drill technique); and (3) a 4.5 mm drill bit used in a 3 drill technique. The CMC joint was disarticulated after drilling, and cartilage and subchondral bone damage evaluated visually and by gross and microradiographic examination using planimetry. RESULTS: Technique 1 produced significantly more damage of the proximal surface, but significantly less to the subchondral bone of the distal surface. Technique 1 produced the most damage to both the articular cartilage and subchondral bone of the total CMC joint than either of the 3 drill tract techniques; however, the difference between techniques 1 and 2 was not significant. Damage from technique 3 was significantly less than that with techniques 1 or 2. CONCLUSIONS: Techniques 1 and 2 produced the most cartilage and subchondral bone damage with technique 2 changes more equally distributed between proximal and distal joint surfaces. CLINICAL RELEVANCE: Technique 1 (fanning) and 2 (5.5 mm 3 drill tracts) may be preferable to achieve arthrodesis of the CMC joint. Morbidity and efficacy of these arthrodesis techniques need to be evaluated in vivo. PMID- 20017859 TI - Carpometacarpal osteoarthritis in thirty-three horses. AB - OBJECTIVE: To describe signalment, clinical, and radiographic changes associated with carpometacarpal osteoarthritis (CMC-OA) and to report long-term outcome. STUDY DESIGN: Case series. ANIMALS: Horses (n=33) with CMC-OA. METHODS: Medical records (1992-2007) of horses diagnosed with CMC-OA were reviewed and signalment, clinical, and radiographic findings retrieved. Owners were contacted for information on the impact of lameness on intended use, response to treatment, progression of lameness, outcome, and owner satisfaction with response to treatment. RESULTS: CMC-OA identified in 39 limbs, occurred predominantly in middle-aged and older Quarter Horses and Arabians, and caused severe lameness that prevented normal use. Characteristic swelling was centered over the 2nd metacarpal bone/2nd carpal bone articulation. Radiographic changes consisted of proliferative new bone, narrowed joint space, and subchondral lysis. Of 20 horses with follow-up, 7 of 14 treated horses were euthanatized within 4 years and 4 of 5 nontreated horses within 3 years. Response to treatment was short lived and considered very poor by most owners. CONCLUSION: CMC-OA seemingly occurs primarily in Quarter Horses and Arabians in our region. Response to conservative treatment is very poor and short-lived with most horses being euthanatized. CLINICAL RELEVANCE: Conventional treatment methods are unsuccessful for treating CMC-OA. PMID- 20017860 TI - Treatment of carpometacarpal osteoarthritis by arthrodesis in 12 horses. AB - OBJECTIVE: To evaluate arthrodesis as a treatment for carpometacarpal joint osteoarthritis (CMC-OA). STUDY DESIGN: Case series. ANIMALS: Horses (n=12) with CMC-OA. METHODS: Arthrodesis was facilitated by insertion and fanning of a drill bit into the CMC joint at several (3-5) locations in 15 limbs. Follow-up radiographs were obtained for 7 horses (9 limbs). Outcome was determined by telephone survey of owners based on postoperative pain, return to use, appearance of the limb, and success of treatment. RESULTS: Postoperative pain was slight or moderate in 10 of 12 (83%) horses during the first 30 days, and 11 of 12 (92%) horses were markedly improved by 6 months and capable of returning to work. Radiographically 6 CMC joints had a bony ankylosis at follow-up whereas 3 did not (mean 8.7 months). On long-term follow-up (mean 28.6 months) all horses had reduced severity of lameness, 10 of 12 (83%) were considered "sound," 8 (67%) returned to their original activity, and all owners considered arthrodesis highly successful as a treatment. CONCLUSION: A drilling technique that produced CMC arthrodesis, allowed most horses to return to their original activity and was considered successful by all clients. CLINICAL RELEVANCE: Arthrodesis of the CMC joint should be considered a treatment option for CMC-OA. PMID- 20017861 TI - Inguinal herniation of the ascending colon in a 6-month-old Standardbred colt. AB - OBJECTIVE: To report inguinal herniation of the ascending colon in a Standardbred colt. STUDY DESIGN: Case report. ANIMALS: A 6-month-old Standardbred colt. METHODS: The colt underwent surgical exploration of the hernia with large colon resection and anastomosis. RESULTS: A successful large colon resection and anastomosis was completed, however, the colt was euthanatized at the end of surgery upon owner request. CONCLUSIONS AND CLINICAL RELEVANCE: Although uncommon, indirect inguinal herniation of the ascending colon can occur in young horses. PMID- 20017862 TI - Video-assisted left partial arytenoidectomy by diode laser photoablation for treatment of canine laryngeal paralysis. PMID- 20017864 TI - Report on the joint IBEPAG/ICCBBA survey on import/export and blood component labelling. AB - A survey of blood centre organizations was carried out to establish the degree of progress towards the implementation of global standards for coding and labelling blood components. The survey was performed through questionnaires completed by blood organizations. Of nearly 32 million blood donations collected annually by the participants, 43% are identified with ISBT 128 donation numbers and 36% are fully compliant with the ISBT 128 Standard. Planned implementations indicate that 85% of donations will be identified by ISBT 128 donation numbers by 2011. PMID- 20017865 TI - Effects of dexmedetomidine sedation on the EEG in children. AB - OBJECTIVES: To examine the effects of dexmedetomidine sedation on EEG background and epileptiform activity in children, comparing it to natural sleep. AIM: To provide quantitative and qualitative descriptions of the effect of dexmedetomidine sedation on the EEG of children. BACKGROUND: Children with intractable epilepsy admitted for surgery undergo 5 days of continuous EEG monitoring as well as nuclear medicine imaging studies with dexmedetomidine for sedation. Continuous EEG monitoring of each child during both natural sleep and dexmedetomidine-induced sedation provides a unique opportunity to evaluate the effects of dexmedetomidine on the EEG of children. MATERIALS/METHODS: Sixteen children undergoing dexmedetomidine sedation for nuclear medicine studies and simultaneous continuous EEG monitoring were studied. EEG segments during sedation were compared to samples of naturally occurring stage II sleep from the same child. Standard visual EEG analysis, quantification of delta, theta, alpha, beta, and total RMS power, number and location of spike foci, and frequency of spike activity were compared. RESULTS: The EEG during dexmedetomidine sedation resembled stage II sleep. During sedation, statistically significant increases in power of 16% for theta (P = 0.01), 21% for alpha (P = 0.03), and 40% for beta (P < 0.01) were observed, but not for delta (P = 0.63) or total EEG power (P = 0.61). Spike frequency increased by 47% during sedation but no new spike foci or seizures were observed. CONCLUSION: Dexmedetomidine sedation elicited an EEG pattern similar to that of Stage II sleep with modest increases in theta, alpha, and beta activity. Dexmedetomidine does not hinder interpretation of the EEG, suggesting that it may be a uniquely useful agent for EEG sedation in children. PMID- 20017866 TI - Presurgical fentanyl vs caudal block and the incidence of adverse respiratory events in children after orchidopexy. AB - BACKGROUND: There is controversy about the etiology of early postoperative hypoxemia. Age, weight, intubation, surgical procedure, use of muscle relaxants, and/or administration of opioids may affect the incidence of early postoperative hypoxemia. In this prospective, randomized, and single-blinded study, we evaluated whether the administration of caudal analgesia vs i.v. fentanyl affected the number of children who develop postextubation adverse upper airway respiratory events, (upper airway obstruction, laryngospasm) and/or early postoperative hypoxemia. METHODS/MATERIALS: Institutional approval and written parental informed consents were obtained. Thirty-eight healthy outpatient boys, aged 1-6 years, scheduled for elective orchidopexy were randomized to receive pain relief either with a presurgical caudal block or by i.v. fentanyl. The primary outcome of the study was the number of children who developed postextubation adverse upper airway respiratory events and/or early postoperative hypoxemia. RESULTS: The number of boys who developed postextubation adverse upper airway respiratory events and/or early postoperative hypoxemia in the caudal group was less compared with those in the fentanyl group (P = 0.04). CONCLUSIONS: Compared to fentanyl, placement of a presurgical caudal block in boys scheduled for orchidopexy was associated with a lower incidence of postextubation adverse upper airway respiratory events and/or early postoperative hypoxemia. PMID- 20017868 TI - Performing US-guided nerve blocks in the postanesthesia care unit (PACU) for upper extremity fractures: is this feasible in children? PMID- 20017869 TI - Changing an oral endotracheal tube to a nasal tube in Pierre-Robin sequence. PMID- 20017870 TI - Pediatric airway bleeding: is rapid-sequence airway control mandatory? PMID- 20017871 TI - A lost art: casualty of deep tracheal extubation. PMID- 20017872 TI - A variation on the modified LMA for fiberoptic intubation. PMID- 20017873 TI - A cost-effective alternative to wire-guided endobronchial blocker for lung isolation in children. PMID- 20017874 TI - Larsen syndrome and malignant hyperthermia. PMID- 20017875 TI - Early transfusion and crystalloid infusion strategy in infants undergoing cranioplasty surgery. PMID- 20017877 TI - Nursing science: knowledge development for the good of persons and society. PMID- 20017878 TI - Professional knowledge and the epistemology of reflective practice. AB - Reflective practice is one of the most popular theories of professional knowledge in the last 20 years and has been widely adopted by nursing, health, and social care professions. The term was coined by Donald Schon in his influential books The Reflective Practitioner, and Educating the Reflective Practitioner, and has garnered the unprecedented attention of theorists and practitioners of professional education and practice. Reflective practice has been integrated into professional preparatory programmes, continuing education programmes, and by the regulatory bodies of a wide range of health and social care professions. Yet, despite its popularity and widespread adoption, a problem frequently raised in the literature concerns the lack of conceptual clarity surrounding the term reflective practice. This paper seeks to respond to this problem by offering an analysis of the epistemology of reflective practice as revealed through a critical examination of philosophical influences within the theory. The aim is to discern philosophical underpinnings of reflective practice in order to advance increasingly coherent interpretations, and to consider the implications for conceptions of professional knowledge in professional life. The paper briefly examines major philosophical underpinnings in reflective practice to explicate central themes that inform the epistemological assumptions of the theory. The study draws on the work of Donald Schon, and on texts from four philosophers: John Dewey, Nelson Goodman, Michael Polanyi, and Gilbert Ryle. Five central epistemological themes in reflective practice are illuminated: (1) a broad critique of technical rationality; (2) professional practice knowledge as artistry; (3) constructivist assumptions in the theory; (4) the significance of tacit knowledge for professional practice knowledge; and (5) overcoming mind body dualism to recognize the knowledge revealed in intelligent action. The paper reveals that the theory of reflective practice is concerned with deep epistemological questions of significance to conceptions of knowledge in health and social care professions. PMID- 20017879 TI - Ontologies of nursing in an age of spiritual pluralism: closed or open worldview? AB - North American society has undergone a period of sacralization where ideas of spirituality have increasingly been infused into the public domain. This sacralization is particularly evident in the nursing discourse where it is common to find claims about the nature of persons as inherently spiritual, about what a spiritually healthy person looks like and about the environment as spiritually energetic and interconnected. Nursing theoretical thinking has also used claims about the nature of persons, health, and the environment to attempt to establish a unified ontology for the discipline. However, despite this common ground, there has been little discussion about the intersections between nursing philosophic thinking and the spirituality in nursing discourse, or about the challenges of adopting a common view of these claims within a spiritually pluralist society. The purpose of this paper is to discuss the call for ontological unity within nursing philosophic thinking in the context of the sacralization of a diverse society. I will begin with a discussion of secularization and sacralization, illustrating the diversity of beliefs and experiences that characterize the current trend towards sacralization. I will then discuss the challenges of a unified ontological perspective, or closed world view, for this diversity, using examples from both a naturalistic and a unitary perspective. I will conclude by arguing for a unified approach within nursing ethics rather than nursing ontology. PMID- 20017880 TI - A response to 'Ontologies of nursing in an age of spiritual pluralism: closed or open worldview?' by Barbara Pesut: our review of the Central Unifying Focus perspective as implying an open worldview: a clarification. PMID- 20017881 TI - Symbolic interactionism and critical perspective: divergent or synergistic? AB - Throughout their history, symbolic interactionism and critical perspective have been viewed as divergent theoretical perspectives with different philosophical underpinnings. A review of their historical and philosophical origins reveals both points of divergence and areas of convergence. Their underlying philosophies of science and views of human freedom are different as is their level of focus with symbolic interactionism having a micro perspective and critical perspective using a macro perspective. This micro/macro difference is reflected in the divergence of their major concepts, goals and basic tenets. While their underlying philosophies are different, however, they are not necessarily contradictory and areas of convergence may include the concepts of reference groups and looking glass self within symbolic interactionism and ideological hegemony within critical perspective. By using a pragmatic approach and combining symbolic interactionism and critical perspectives, both micro and macro levels come into focus and strategies for change across individual and societal levels can be developed and applied. Application of both symbolic interactionism and critical perspective to nursing research and scholarship offers exciting new opportunities for theory development and research methodologies. In nursing education, these two perspectives can give students added insight into patients' and families' problems at the micro level while, at the same time, giving them a lens to see and tools to apply to problems at the macro level in health care. In nursing practice, a combined symbolic interactionism/critical perspective approach assists nurses to give high-quality care at the individual level while also working at the macro level to address the manufacturers of illness. New research questions emerge from this combination of perspectives with new possibilities for theory development, a transformation in nursing education, and the potential for new practice strategies that can address individual client and larger system problems through empowerment of clients and nurses. PMID- 20017882 TI - Knowledge for the good of the individual and society: linking philosophy, disciplinary goals, theory, and practice. AB - Nursing as a profession has a social mandate to contribute to the good of society through knowledge-based practice. Knowledge is built upon theories, and theories, together with their philosophical bases and disciplinary goals, are the guiding frameworks for practice. This article explores a philosophical perspective of nursing's social mandate, the disciplinary goals for the good of the individual and society, and one approach for translating knowledge into practice through the use of a middle-range theory. It is anticipated that the integration of the philosophical perspective and model into nursing practice will strengthen the philosophy, disciplinary goal, theory, and practice links and expand knowledge within the discipline. With the focus on humanization, we propose that nursing knowledge for social good will embrace a synthesis of the individual and the common good. This approach converges vital and agency needs described by Hamilton and the primacy of maintaining the heritage of the good within the human species as outlined by Maritain. Further, by embedding knowledge development in a changing social and health care context, nursing focuses on the goals of clinical reasoning and action. McCubbin and Patterson's Double ABCX Model of Family Adaptation was used as an example of a theory that can guide practice at the community and global level. Using the theory-practice link as a foundation, the Double ABCX model provides practising nurses with one approach to meet the needs of individuals and society. The integration of theory into nursing practice provides a guide to achieve nursing's disciplinary goals of promoting health and preventing illness across the globe. When nursing goals are directed at the synthesis of the good of the individual and society, nursing's social and moral mandate may be achieved. PMID- 20017883 TI - A model of consensus formation for reconciling nursing's disciplinary matrix. AB - With questions raised as to whether or not nursing knowledge should be developed from extant conceptual/theoretical models or from practice-based environments, this paper utilizes Kuhn's disciplinary matrix and Laudan's model of consensus formation to explore the changing nature of the discipline's structural matrix. Kuhn's notion that a discipline's structural matrix includes symbolic generalizations, models and exemplars, and Laudan's view that a maturing discipline embraces factual, methodological, and axiological (goals and aims) knowledge, and that context and discourse are also involved in advancing a discipline is described as a means for reconciling the source of nursing knowledge. This paper posits that shared axiological goals connect both theorists and practitioners, and resolve potential conflicts as to viable sources of nursing knowledge. Through shared goals that include humanization, meaning, quality of life, caring, consciousness, transcendence, and presence, which bridge both theoretical and practice approaches, nursing's charge to contribute to the good of society is fulfilled. PMID- 20017884 TI - A nursing manifesto: an emancipatory call for knowledge development, conscience, and praxis. AB - The purpose of this paper is to present the theoretical and philosophical assumptions of the Nursing Manifesto, written by three activist scholars whose objective was to promote emancipatory nursing research, practice, and education within the dialogue and praxis of social justice. Inspired by discussions with a number of nurse philosophers at the 2008 Knowledge Conference in Boston, two of the original Manifesto authors and two colleagues discussed the need to explicate emancipatory knowing as it emerged from the Manifesto. Our analysis yielded an epistemological framework based on liberation principles to advance praxis in the discipline of nursing. This paper adds to what is already known on this topic, as there is not an explicit contribution to the literature of this specific Manifesto, its significance, and utility for the discipline. While each of us have written on emancipatory knowing and social justice in a variety of works, it is in this article that we identify, as a unit of knowledge production and as a direction towards praxis, a set of critical values that arose from the emancipatory conscience-ness and intention seen in the framework of the Nursing Manifesto. PMID- 20017885 TI - PARP inhibitors and the treatment of breast cancer: beyond BRCA1/2? AB - Poly(ADP-ribose) polymerase (PARP) inhibitors have been explored as therapeutic agents for the treatment of hereditary breast and ovarian cancers harboring mutations in BRCA1 or BRCA2. In a new study, Inbar-Rozensal and colleagues show that phenanthridine-derived PARP inhibitors promote cell cycle arrest and cell death in breast cancer cells lacking BRCA1 and BRCA2 mutations and prevent the growth of tumors from xenografts of these cells in immunocompromised mice. These results suggest a potential broader utility of PARP-1 inhibitors in the treatment of breast cancer, although further mechanistic studies are needed. PMID- 20017886 TI - Role of chlorhexidine gluconate in ventilator-associated pneumonia prevention strategies in ICU patients: where are we headed? PMID- 20017887 TI - Adaptations of proteins to cellular and subcellular pH. AB - Bioinformatics-based searches for correlations between subcellular localization and pI or charge distribution of proteins have failed to detect meaningful correlations. Recent work published in BMC Biology finds that a physicochemical metric of charge distribution correlates better with subcellular pH than does pI. See research article http://www.biomedcentral.com/1741-7007/7/69. PMID- 20017888 TI - Rituximab treatment in rheumatoid arthritis: how does it work? AB - Treatment with the chimerical monoclonal antibody rituximab results in CD20 directed B cell depletion. Although this depletion is almost complete in the peripheral blood of nearly all patients with rheumatoid arthritis, a proportion of patients does not exhibit a clinical response. The paper by Nakou and colleagues suggests that a decrease in CD19+CD27+ memory B cells in both peripheral blood and bone marrow precedes the clinical response to rituximab. This finding adds to the emerging evidence that lack of response to rituximab is associated with persistence of B lineage cells in specific body compartments. PMID- 20017889 TI - Clinical review: a review and analysis of heart rate variability and the diagnosis and prognosis of infection. AB - Bacterial infection leading to organ failure is the most common cause of death in critically ill patients. Early diagnosis and expeditious treatment is a cornerstone of therapy. Evaluating the systemic host response to infection as a complex system provides novel insights: however, bedside application with clinical value remains wanting. Providing an integrative measure of an altered host response, the patterns and character of heart rate fluctuations measured over intervals-in-time may be analysed with a panel of mathematical techniques that quantify overall fluctuation, spectral composition, scale-free variation, and degree of irregularity or complexity. Using these techniques, heart rate variability (HRV) has been documented to be both altered in the presence of systemic infection, and correlated with its severity. In this review and analysis, we evaluate the use of HRV monitoring to provide early diagnosis of infection, document the prognostic implications of altered HRV in infection, identify current limitations, highlight future research challenges, and propose improvement strategies. Given existing evidence and potential for further technological advances, we believe that longitudinal, individualized, and comprehensive HRV monitoring in critically ill patients at risk for or with existing infection offers a means to harness the clinical potential of this bedside application of complex systems science. PMID- 20017890 TI - Matrix metalloproteinases and their inhibitors: promising novel biomarkers in severe sepsis? AB - The multicenter study conducted by Lorente and coworkers published in the previous issue of Critical Care demonstrates that matrix metalloproteinase (MMP) 9 and MMP-10 and their inhibitor tissue inhibitor of matrix metalloproteinase-1 (TIMP-1) are promising novel biomarkers to predict severity and outcome of sepsis. In recent years MMPs have emerged as biomarkers in a variety of diseases, such as sepsis, coronary artery disease, cancer, heart failure, chronic lung disease and rheumatoid arthritis. MMPs constitute a family of proteinases that are expressed during developmental, physiological, and pathophysiological processes, for example as a response to infection. Excessive inflammation following infection may cause tissue damage, and MMPs are implicated in causing this immunopathology. The activity of MMPs is regulated by secretion of specific inhibitors (TIMPs). Studies using MMP inhibitors and MMP knockout mice indicate that MMPs play an essential role in infection and in the host response to infection. The measurement of MMP-9 and MMP-10 and their inhibitor TIMP-1 in the intensive care setting could be an attractive noninvasive tool for determination of outcome of septic patients. PMID- 20017891 TI - Clinical review: post-extubation laryngeal edema and extubation failure in critically ill adult patients. AB - Laryngeal edema is a frequent complication of intubation. It often presents shortly after extubation as post-extubation stridor and results from damage to the mucosa of the larynx. Mucosal damage is caused by pressure and ischemia resulting in an inflammatory response. Laryngeal edema may compromise the airway necessitating reintubation. Several studies show that a positive cuff leak test combined with the presence of risk factors can identify patients with increased risk for laryngeal edema. Meta-analyses show that pre-emptive administration of a multiple-dose regimen of glucocorticosteroids can reduce the incidence of laryngeal edema and subsequent reintubation. If post-extubation edema occurs this may necessitate medical intervention. Parenteral administration of corticosteroids, epinephrine nebulization and inhalation of a helium/oxygen mixture are potentially effective, although this has not been confirmed by randomized controlled trials. The use of non-invasive positive pressure ventilation is not indicated since this will delay reintubation. Reintubation should be considered early after onset of laryngeal edema to adequately secure an airway. Reintubation leads to increased cost, morbidity and mortality. PMID- 20017892 TI - Musings on genome medicine: enzyme-replacement therapy of the lysosomal storage diseases. AB - The lysosomal storage diseases, such as Gaucher's disease, mucopolysaccharidosis I, II and IV, Fabry's disease, and Pompe's disease, are rare inherited disorders whose symptoms result from enzyme deficiency causing lysosomal accumulation. Until effective gene-replacement therapy is developed, expensive, and at best incomplete, enzyme-replacement therapy is the only hope for sufferers of rare lysosomal storage diseases. Preventive strategies involving carrier detection should be a priority toward the successful management of these conditions. PMID- 20017893 TI - Host immune response in sepsis due to ventilator-associated pneumonia: how is it different? AB - Current evidence regarding potentially different host response mechanisms in sepsis according to the type of initiating infection is sporadic. It is possible that alterations in cell populations, variations in effector molecules, and the degree of apoptosis differ between sepsis caused by ventilator-associated pneumonia (VAP) and non-VAP sepsis. VAP is one of the most common infections and leading causes of sepsis in the intensive care unit, and mortality remains high. A better understanding of the unique pathophysiologic features of VAP is needed in order to develop interventions that target those specific pathways. PMID- 20017894 TI - The propofol infusion syndrome: more puzzling evidence on a complex and poorly characterized disorder. AB - The propofol infusion syndrome is a potentially devastating cardiovascular and metabolic derangement that has been described in both pediatric and adult patients sedated with propofol. Despite a large number of case reports that have appeared in the literature since 1992, the precise clinical features and pathophysiology of this disorder remain uncertain. Historically, the syndrome has been characterized by the occurrence of lactic acidosis, rhabdomyolysis, and circulatory collapse after several days of high-dose propofol infusion. The affected patients were typically young and critically ill, and the reported mortality was high. More recently, a number of atypical cases have been reported with favorable outcomes. These occurred after short-term or lower-dose infusions in noncritically ill patients in whom generally only a subset of the classical syndrome features was observed. It remains unclear whether these reports reflect true propofol infusion syndrome detected at an earlier and more salvageable stage, or mere associations with the use of sedative agents in general. Without better information on the true incidence of the propofol infusion syndrome, clinical guidelines on the safe use of this drug remain unsupported by good evidence. PMID- 20017895 TI - Death by acid rain: VAP or EXIT? AB - Ventilator-associated pneumonia (VAP) is a new (nosocomial) lower respiratory tract infection diagnosed in mechanically ventilated patients 48 or more hours after intubation. There is no gold standard for establishing the diagnosis and its pathogenesis is iatrogenic and multifactorial. Gastro-oesophageal reflux is common in mechanically ventilated children, but its role in VAP remains speculative. VAP is associated with increased mortality and morbidity, prolonged duration of ventilation and hospital stay, and escalated costs of hospitalisation. VAP 'bundles' are championed as the antidote. PMID- 20017896 TI - Coordinated gene expression by post-transcriptional regulons in African trypanosomes. AB - The regulation of gene expression in trypanosomes is unique. In the absence of transcriptional control at the level of initiation, a subset of Trypanosoma brucei genes form post-transcriptional regulons in which mRNAs are co-regulated in response to differentiation signals. See research articles http://www.biomedcentral.com/1471-2164/10/427, http://www.biomedcentral.com/1471 2164/10/482 and http://www.biomedcentral.com/1471-2164/10/495. PMID- 20017897 TI - Promoter architecture and the evolvability of gene expression. AB - Evolutionary changes in gene expression are a main driver of phenotypic evolution. In yeast, genes that have rapidly diverged in expression are associated with particular promoter features, including the presence of a TATA box, a nucleosome-covered promoter and unstable tracts of tandem repeats. Here, we discuss how these promoter properties may confer an inherent capacity for flexibility of expression. PMID- 20017898 TI - beta2 adrenergic agonists in acute lung injury? The heart of the matter. AB - Despite extensive research into its pathophysiology, acute lung injury/acute respiratory distress syndrome (ALI/ARDS) remains a devastating syndrome with mortality approaching 40%. Pharmacologic therapies that reduce the severity of lung injury in vivo and in vitro have not yet been translated to effective clinical treatment options, and innovative therapies are needed. Recently, the use of beta2 adrenergic agonists as potential therapy has gained considerable interest due to their ability to increase the resolution of pulmonary edema. However, the results of clinical trials of beta agonist therapy for ALI/ARDS have been conflicting in terms of benefit. In the previous issue of Critical Care, Briot and colleagues present evidence that may help clarify the inconsistent results. The authors demonstrate that, in oleic acid lung injury in dogs, the inotropic effect of beta agonists may recruit damaged pulmonary capillaries, leading to increased lung endothelial permeability. PMID- 20017899 TI - Statin research in critical illness: hampered by poor trial design? AB - Statin therapy may prevent an excessive inflammatory response after cardiopulmonary bypass for cardiac surgery. In a recent issue of Critical Care, Morgan and colleagues present data from a well-conducted systematic review and meta-analysis of randomised controlled trials using inflammatory markers as primary outcome measure. They find that pre-operative statin therapy, compared with placebo, may reduce various post-operative markers of systemic inflammation (IL-6, IL-8, C-reactive protein, tumour necrosis factor-alpha). Their ability to make definitive conclusions is limited, however, by the suboptimal methodological quality of the primary studies. Their review suggests that ICU researchers should focus on developing valid surrogate markers and use these to accurately describe the mechanisms and effectiveness of novel therapies before proceeding to large pragmatic trials using mortality as primary outcome. PMID- 20017900 TI - The role of anti-alpha-actinin antibodies in the pathogenesis and monitoring of lupus nephritis. AB - Antibodies to double-stranded DNA are important in the pathogenesis of nephritis, a major clinical manifestation in lupus patients. Since earlier diagnosis of renal involvement may lead to better outcomes, identification of the nephritogenic specificity of lupus-associated autoantibodies is important in understanding the disease, while monitoring their titer clinically may serve as an improved biomarker. Based upon work in animal models and cross-sectional human studies, kidney alpha-actinin was thought to be a plausible cross-reactive target for pathogenic lupus antibodies. Manson and colleagues longitudinally evaluated anti-nucleosome, anti-DNA, and anti-alpha-actinin antibodies in 16 lupus patients with new-onset nephritis. While anti-nucleosome and anti-DNA antibody levels were significantly associated and correlated with measures of kidney disease, these were not found to be significant with anti-alpha-actinin antibodies. While in lupus patients the diagnostic use of serum anti-alpha-actinin antibodies, alone or with other novel biomarkers, is still under investigation, such studies are vital in improving our monitoring of systemic lupus erythematosus patients and in developing new treatment paradigms that meet the continuing clinical challenge of lupus nephritis. PMID- 20017901 TI - A novel anti-mycobacterial function of mitogen-activated protein kinase phosphatase-1. AB - BACKGROUND: Mycobacterium tuberculosis (MTB) is a major cause of morbidity and mortality in the world. To combat against this pathogen, immune cells release cytokines including tumor necrosis factor-alpha (TNF-alpha), which is pivotal in the development of protective granulomas. Our previous results showed that Bacillus Calmette Guerin (BCG), a mycobacterium used as a model to investigate the immune response against MTB, stimulates the induction of TNF-alpha via mitogen-activated protein kinase (MAPK) in human blood monocytes. Since MAPK phosphatase-1 (MKP-1) is known to regulate MAPK activities, we examined whether MKP-1 plays a role in BCG-induced MAPK activation and cytokine expression. RESULTS: Primary human blood monocytes were treated with BCG and assayed for MKP 1 expression. Our results demonstrated that following exposure to BCG, there was an increase in the expression of MKP-1. Additionally, the induction of MKP-1 was regulated by p38 MAPK and extracellular signal-regulated kinase 1 and 2 (ERK1/2). Surprisingly, when MKP-1 expression was blocked by its specific siRNA, there was a significant decrease in the levels of phospho-MAPK (p38 MAPK and ERK1/2) and TNF-alpha inducible by BCG. CONCLUSIONS: Since TNF-alpha is pivotal in granuloma formation, the results indicated an unexpected positive function of MKP-1 against mycobacterial infection as opposed to its usual phosphatase activity. PMID- 20017902 TI - Interleukin-23 is critical for full-blown expression of a non-autoimmune destructive arthritis and regulates interleukin-17A and RORgammat in gammadelta T cells. AB - INTRODUCTION: Interleukin (IL)-23 is essential for the development of various experimental autoimmune models. However, the role of IL-23 in non-autoimmune experimental arthritis remains unclear. Here, we examined the role of IL-23 in the non-autoimmune antigen-induced arthritis (AIA) model. In addition, the regulatory potential of IL-23 in IL-17A and retinoic acid-related orphan receptor gamma t (RORgammat) expression in CD4+ and TCRgammadelta+ T cells was evaluated systemically as well as at the site of inflammation. METHODS: Antigen-induced arthritis was induced in wild-type, IL-23p19-deficient and IL-17 Receptor A - knockout mice. At different time points, synovial cytokine and chemokine expression was measured. At days 1 and 7 of AIA, splenocytes and joint infiltrating cells were isolated and analyzed for intracellular IL-17A and interferon (IFN)-gamma ex-vivo by flow cytometry. In splenic CD4+ and TCRgammadelta+ T cells gene expression was quantified by flow cytometry and quantitative PCR. RESULTS: IL-23 was critical for full-blown AIA. Lack of IL-23 did not prevent the onset of joint inflammation but stopped the progression to a destructive synovitis. IL-23 regulated IL-17A expression in CD4+ T cells in the spleen. Of note, IL-17A and IFN-gamma expression was reduced in CD4+ T cells in the inflamed joints of IL-23p19-deficient mice. Interestingly, IL-23 was also critical for the induction of IL-17A and RORgammat but not IFN-gamma in TCRgammadelta+ T cells in the inflamed joints. The importance of the IL-23/IL-17 axis was further confirmed using IL-17 Receptor A knockout mice showing significantly milder AIA compared to control mice, with a disease course comparable to that of IL-23p19-deficient mice. CONCLUSIONS: These data show that IL-23 is critical for full-blown expression of a non-autoimmune destructive arthritis and regulates the proportion of IL-17A and IFN-gamma-positive CD4+ T cells at the site of inflammation. Furthermore, IL-23 regulates IL-17A and RORgammat expression in TCRgammadelta T cells in arthritis. These findings indicate that regulating the IL-23 pathway may have therapeutic potential in non autoimmune arthritis. PMID- 20017903 TI - Melusin gene (ITGB1BP2) nucleotide variations study in hypertensive and cardiopathic patients. AB - BACKGROUND: Melusin is a muscle specific signaling protein, required for compensatory hypertrophy response in pressure-overloaded heart. The role of Melusin in heart function has been established both by loss and gain of function experiments in murine models. With the aim of verifying the hypothesis of a potential role of the Melusin encoding gene, ITGB1BP2, in the modification of the clinical phenotype of human cardiomyopathies, we screened the ITGB1BP2 gene looking for genetic variations possibly associated to the pathological phenotype in three selected groups of patients affected by hypertension and dilated or hypertrophic cardiomyopathy METHODS: We analyzed ITGB1BP2 by direct sequencing of the 11 coding exons and intron flanking sequences in 928 subjects, including 656 hypertensive or cardiopathic patients and 272 healthy individuals. RESULTS: Only three nucleotide variations were found in patients of three distinct families: a C>T missense substitution at position 37 of exon 1 causing an amino acid change from His-13 to Tyr in the protein primary sequence, a duplication (IVS6+12_18dupTTTTGAG) near the 5'donor splice site of intron 6, and a silent 843C>T substitution in exon 11. CONCLUSIONS: The three variations of the ITGB1BP2 gene have been detected in families of patients affected either by hypertension or primary hypertrophic cardiomyopathy; however, a clear genotype/phenotype correlation was not evident. Preliminary functional results and bioinformatic analysis seem to exclude a role for IVS6+12_18dupTTTTGAG and 843C>T in affecting splicing mechanism.Our analysis revealed an extremely low number of variations in the ITGB1BP2 gene in nearly 1000 hypertensive/cardiopathic and healthy individuals, thus suggesting a high degree of conservation of the melusin gene within the populations analyzed. PMID- 20017904 TI - Upstream ORF affects MYCN translation depending on exon 1b alternative splicing. AB - BACKGROUND: The MYCN gene is transcribed into two major mRNAs: one full-length (MYCN) and one exon 1b-spliced (MYCNDelta1b) mRNA. But nothing is known about their respective ability to translate the MYCN protein. METHODS: Plasmids were prepared to enable translation from the upstream (uORF) and major ORF of the two MYCN transcripts. Translation was studied after transfection in neuroblastoma SH EP cell line. Impact of the upstream AUG on translation was evaluated after directed mutagenesis. Functional study with the two MYCN mRNAs was conducted by a cell viability assay. Existence of a new protein encoded by the MYCNDelta1b uORF was explored by designing a rabbit polyclonal antibody against a specific epitope of this protein. RESULTS: Both are translated, but higher levels of protein were seen with MYCNDelta1b mRNA. An upstream ORF was shown to have positive cis regulatory activity on translation from MYCN but not from MYCNDelta1b mRNA. In transfected SH-EP neuroblastoma cells, high MYCN dosage obtained with MYCNDelta1b mRNA translation induces an antiapoptotic effect after serum deprivation that was not observed with low MYCN expression obtained with MYCN mRNA. Here, we showed that MYCNOT: MYCN Overlap Transcript, a new protein of unknown function is translated from the upstream AUG of MYCNDelta1b mRNA. CONCLUSIONS: Existence of upstream ORF in MYCN transcripts leads to a new level of MYCN regulation. The resulting MYCN dosage has a weak but significant anti-apoptotic activity after intrinsic apoptosis induction. PMID- 20017905 TI - Evidence of decline of malaria in the general hospital of Libreville, Gabon from 2000 to 2008. AB - BACKGROUND: Substantial decline in malaria transmission, morbidity and mortality has been reported in several countries where new malaria control strategies have been implemented. In Gabon, the national malaria policy changed in 2003, according to the WHO recommendations. The trend in malaria morbidity was evaluated among febrile children before and after their implementation in Libreville, the capital city of Gabon. METHODS: From August 2000 to December 2008, febrile paediatric outpatients and inpatients, under 11 years of age, were screened for malaria by microscopic examination at the Malaria Clinical Research Unit (MCRU) located in the largest public hospital in Gabon. Climatic data were also collected. RESULTS: In total, 28,092 febrile children were examined; those under five years always represented more than 70%. The proportion of malaria positive slides was 45% in 2000, and declined to 15% in 2008. The median age of children with a positive blood smear increased from 24(15-48) to 41(21-72) months over the study period (p < 0.01). Rainfall patterns had no impact on the decline observed throughout the study period. CONCLUSION: The decrease of malaria prevalence among febrile children during the last nine years is observed following the introduction of new strategies of malaria cases management, and may announce epidemiological changes. Moreover, preventive measures must be extended to children older than five years. PMID- 20017906 TI - DDIT3/CHOP and the sarcoma fusion oncoprotein FUS-DDIT3/TLS-CHOP bind cyclin dependent kinase 2. AB - BACKGROUND: The DDIT3 gene encodes a transcription factor belonging to the CCAAT/enhancer binding protein (C/EBP) family. It is normally expressed at very low levels but is activated by cellular stress conditions and induces G1 arrest and, in some cell types, apoptosis. DDIT3 is found as a part of the fusion oncogene FUS-DDIT3 that is causal for the development of myxoid/round-cell liposarcomas (MLS/RCLS). RESULTS: In the present study, we searched for putative interaction partners of DDIT3 and the oncogenic FUS-DDIT3 among G1 cyclins and cyclin-dependent kinases. We found that FUS-DDIT3 and the normal DDIT3 bind CDK2. In addition, CDK2 showed an increased affinity for cytoskeletal proteins in cells expressing FUS-DDIT3 and DDIT3. CONCLUSIONS: We conclude that DDIT3 binds CDK2 and that many of the observed biological effects of DDIT3 may involve interaction with CDK2. PMID- 20017907 TI - Individual-level socioeconomic status is associated with worse asthma morbidity in patients with asthma. AB - BACKGROUND: Low socioeconomic status (SES) has been linked to higher morbidity in patients with chronic diseases, but may be particularly relevant to asthma, as asthmatics of lower SES may have higher exposures to indoor (e.g., cockroaches, tobacco smoke) and outdoor (e.g., urban pollution) allergens, thus increasing risk for exacerbations. METHODS: This study assessed associations between adult SES (measured according to educational level) and asthma morbidity, including asthma control; asthma-related emergency health service use; asthma self efficacy, and asthma-related quality of life, in a Canadian cohort of 781 adult asthmatics. All patients underwent a sociodemographic and medical history interview and pulmonary function testing on the day of their asthma clinic visit, and completed a battery of questionnaires (Asthma Control Questionnaire, Asthma Quality of Life Questionnaire, and Asthma Self-Efficacy Scale). General Linear Models assessed associations between SES and each morbidity measure. RESULTS: Lower SES was associated with worse asthma control (F = 11.63, p < .001), greater emergency health service use (F = 5.09, p = .024), and worse asthma self-efficacy (F = 12.04, p < .01), independent of covariates. Logistic regression analyses revealed that patients with <12 years of education were 55% more likely to report an asthma-related emergency health service visit in the last year (OR = 1.55, 95%CI = 1.05-2.27). Lower SES was not related to worse asthma-related quality of life. CONCLUSIONS: Results suggest that lower SES (measured according to education level), is associated with several indices of worse asthma morbidity, particularly worse asthma control, in adult asthmatics independent of disease severity. Results are consistent with previous studies linking lower SES to worse asthma in children, and add asthma to the list of chronic diseases affected by individual-level SES. PMID- 20017908 TI - Hyperoxia increases the uptake of 5-fluorouracil in mammary tumors independently of changes in interstitial fluid pressure and tumor stroma. AB - BACKGROUND: Hypoxia is associated with increased resistance to chemo- and radiation-therapy. Hyperoxic treatment (hyperbaric oxygen) has previously been shown to potentiate the effect of some forms of chemotherapy, and this has been ascribed to enhanced cytotoxicity or neovascularisation. The aim of this study was to elucidate whether hyperoxia also enhances any actual uptake of 5FU (5 fluorouracil) into the tumor tissue and if this can be explained by changes in the interstitium and extracellular matrix. METHODS: One group of tumor bearing rats was exposed to repeated hyperbaric oxygen (HBO) treatment (2 bar, pO(2)= 2 bar, 4 exposures a 90 min), whereas one group was exposed to one single identical HBO treatment. Animals housed under normal atmosphere (1 bar, pO(2) = 0.2 bar) served as controls. Three doses of 5FU were tested for dose response. Uptake of [3H]-5FU in the tumor was assessed, with special reference to factors that might have contributed, such as interstitial fluid pressure (Pif), collagen content, oxygen stress (measured as malondialdehyd levels), lymphatics and transcapillary transport in the tumors. RESULTS: The uptake of the cytostatic agent increases immediately after a single HBO treatment (more than 50%), but not 24 hours after the last repeated HBO treatment. Thus, the uptake is most likely related to the transient increase in oxygenation in the tumor tissue. Factors like tumor Pif and collagen content, which decreased significantly in the tumor interstitium after repeated HBO treatment, was without effect on the drug uptake. CONCLUSION: We showed that hyperoxia increases the uptake of [3H]-5FU in DMBA-induced mammary tumors per se, independently of changes in Pif, oxygen stress, collagen fibril density, or transendothelial transport alone. The mechanism by which such an uptake occur is still not elucidated, but it is clearly stimulated by elevated pO(2). PMID- 20017909 TI - Treatment-seeking for febrile illness in north-east India: an epidemiological study in the malaria endemic zone. AB - BACKGROUND: This paper studies the determinants of utilization of health care services, especially for treatment of febrile illness in the malaria endemic area of north-east India. METHODS: An area served by two districts of Upper Assam representing people living in malaria endemic area was selected for household survey. A sample of 1,989 households, in which at least one member of household suffered from febrile illness during last three months and received treatment from health service providers, were selected randomly and interviewed by using the structured questionnaire. The individual characteristics of patients including social indicators, area of residence and distance of health service centers has been used to discriminate or group the patients with respect to their initial and final choice of service providers. RESULTS: Of 1,989 surveyed households, initial choice of treatment-seeking for febrile illness was self medication (17.8%), traditional healer (Vaidya)(39.2%), government (29.3%) and private (13.7%) health services. Multinomial logistic regression (MLR) analysis exhibits the influence of occupation, area of residence and ethnicity on choice of health service providers. The traditional system of medicine was commonly used by the people living in remote areas compared with towns. As all the febrile cases finally received treatment either from government or private health service providers, the odds (Multivariate Rate Ratio) was almost three-times higher in favour of government services for lower households income people compared to private. CONCLUSION: The study indicates the popular use of self-medication and traditional system especially in remote areas, which may be the main cause of delay in diagnosis of malaria. The malaria training given to the paramedical staff to assist the health care delivery needs to be intensified and expanded in north-east India. The people who are economically poor and living in remote areas mainly visit the government health service providers for seeking treatment. So, the improvement of quality health services in government health sector and provision of health education to people would increase the utilization of government health services and thereby improve the health quality of the people. PMID- 20017910 TI - Comparison of the efficacy of administering a combination of ezetimibe plus fenofibrate versus atorvastatin monotherapy in the treatment of dyslipidemia. AB - BACKGROUND: This trial compares the efficacy of administering a combination of ezetimibe plus fenofibrate as an alternative to statin monotherapy for the treatment of dyslipidemia. In this randomized, unblinded crossover study, 43 patients with documented hypercholesterolemia requiring pharmacotherapy were randomized to receive six weeks of either a combination of 10 mg of ezetimibe plus 160 mg of fenofibrate (combination) or 10 mg of atorvastatin monotherapy (atorvastatin). The primary endpoint was the percentage reduction of low-density lipoprotein cholesterol (LDL-C). RESULTS: LDL-C decreased by 34.6% with the combination therapy versus 36.7% with atorvastatin monotherapy. The difference between the two groups was not statistically significant (p = 0.46). Both study interventions provided similar improvements in total cholesterol (-25.1% with combination versus -24.6% with atorvastatin, p = 0.806) and high-density lipoproteins (+10.0% with combination versus +8.9% with atorvastatin, p = 0.778). Combination therapy showed a trend towards a greater reduction in triglycerides ( 25.4% with combination versus -14.5% with atorvastatin, p = 0.079), although there was no significant difference between the two study interventions in terms of the improvement in the TC:HDL ratio (-29.0% with combination versus -28.7% with atorvastatin, p = 0.904). CONCLUSIONS: The combination of ezetimibe plus fenofibrate appeared to produce nearly identical alterations in serum lipoprotein levels when compared to monotherapy with 10 mg of atorvastatin. Daily treatment with the combination of ezetimibe plus fenofibrate is an acceptable alternative to atorvastatin for the treatment of dyslipidemia in patients who are intolerant of statins. PMID- 20017911 TI - The obesity and inflammatory marker haptoglobin attracts monocytes via interaction with chemokine (C-C motif) receptor 2 (CCR2). AB - BACKGROUND: Obesity is a chronic low inflammatory state. In the obesity condition the white adipose tissue (WAT) is massively infiltrated with monocytes/macrophages, and the nature of the signals recruiting these inflammatory cells has yet to be fully elucidated. Haptoglobin (Hp) is an inflammatory marker and its expression is induced in the WAT of obese subjects. In an effort to elucidate the biological significance of Hp presence in the WAT and of its upregulation in obesity we formulated the hypothesis that Hp may serve as a macrophage chemoattractant. RESULTS: We demonstrated by chemotaxis assay that Hp is able to attract chemokine (C-C motif) receptor 2 (CCR2)-transfected pre-B lymphocytes and monocytes in a dose-dependent manner. Moreover, Hp-mediated migration of monocytes is impaired by CCR2-specific inhibition or previous cell exposure to monocyte chemoattractant protein 1 (MCP1) (also known as CCR2 ligand or chemokine (C-C motif) ligand 2 (CCL2)). Downstream effects of Hp/CCR2 interaction were also investigated: flow cytometry proved that monocytes treated with Hp show reduced CCR2 expression on their surface; Hp interaction induces calcium release that is reduced upon pretreatment with CCR2 antagonist; extracellular signal-regulated kinase (ERK)1/2, a signal transducer activated by CCR2, is phosphorylated following Hp treatment and this phosphorylation is reduced when cells are pretreated with a specific CCR2 inhibitor. Consistently, blocking the ERK1/2 pathway with U0126, the selective inhibitor of the ERK upstream mitogen-activated protein (MAP)-ERK kinase (MEK), results in a dramatic reduction (by almost 100%) of the capability of Hp to induce monocyte migration. CONCLUSIONS: Our data show that Hp is a novel monocyte chemoattractant and that its chemotactic potential is mediated, at least in part. by its interaction with CCR2. PMID- 20017913 TI - Pre-treatment preferences and characteristics among patients seeking in vitro fertilisation. AB - BACKGROUND: This study sought to describe patient features before beginning fertility treatment, and to ascertain their perceptions relative to risk of twin pregnancy outcomes associated with such therapy. METHODS: Data on readiness for twin pregnancy outcome from in vitro fertilisation (IVF) was gathered from men and women before initiating fertility treatment by anonymous questionnaire. RESULTS: A total of 206 women and 204 men were sampled. Mean (+/- SD) age for women and men being 35.5 +/- 5 and 37.3 +/- 7 yrs, respectively. At least one IVF cycle had been attempted by 27.2% of patients and 33.9% of this subgroup had initiated >/=3 cycles, reflecting an increase in previous failed cycles over five years. Good agreement was noted between husbands and wives with respect to readiness for twins from IVF (77% agreement; Cohen's K = 0.61; 95% CI 0.53 to 0.70). CONCLUSION: Most patients contemplating IVF already have ideas about particular outcomes even before treatment begins, and suggests that husbands & wives are in general agreement on their readiness for twin pregnancy from IVF. However, fertility patients now may represent a more refractory population and therefore carry a more guarded prognosis. Patient preferences identified before IVF remain important, but further studies comparing pre- and post-treatment perceptions are needed. PMID- 20017912 TI - Intracellular diffusion restrictions in isolated cardiomyocytes from rainbow trout. AB - BACKGROUND: Restriction of intracellular diffusion of adenine nucleotides has been studied intensively on adult rat cardiomyocytes. However, their cause and role in vivo is still uncertain. Intracellular membrane structures have been suggested to play a role. We therefore chose to study cardiomyocytes from rainbow trout (Oncorhynchus mykiss), which are thinner and have fewer intracellular membrane structures than adult rat cardiomyocytes. Previous studies suggest that trout permeabilized cardiac fibers also have diffusion restrictions. However, results from fibers may be affected by incomplete separation of the cells. This is avoided when studying permeabilized, isolated cardiomyocytes. The aim of this study was to verify the existence of diffusion restrictions in trout cardiomyocytes by comparing ADP-kinetics of mitochondrial respiration in permeabilized fibers, permeabilized cardiomyocytes and isolated mitochondria from rainbow trout heart. Experiments were performed at 10, 15 and 20 degrees C in the absence and presence of creatine. RESULTS: Trout cardiomyocytes hypercontracted in the solutions used for mammalian cardiomyocytes. We developed a new solution in which they retained their shape and showed stable steady state respiration rates throughout an experiment. The apparent ADP-affinity of permeabilized cardiomyocytes was different from that of fibers. It was higher, independent of temperature and not increased by creatine. However, it was still about ten times lower than in isolated mitochondria. CONCLUSIONS: The differences between fibers and cardiomyocytes suggest that results from trout heart fibers were affected by incomplete separation of the cells. However, the lower ADP-affinity of cardiomyocytes compared to isolated mitochondria indicate that intracellular diffusion restrictions are still present in trout cardiomyocytes despite their lower density of intracellular membrane structures. The lack of a creatine effect indicates that trout heart lacks mitochondrial creatine kinase tightly coupled to respiration. This argues against diffusion restriction by the outer mitochondrial membrane. These results from rainbow trout cardiomyocytes resemble those from other low-performance hearts such as neonatal rat and rabbit hearts. Thus, it seems that metabolic regulation is related to cardiac performance, and it is likely that rainbow trout can be used as a model animal for further studies of the localization and role of diffusion restrictions in low-performance hearts. PMID- 20017914 TI - ACL reconstruction with unicondylar replacement in knee with functional instability and osteoarthritis. AB - Severe symptomatic osteoarthritis in young and active patients with pre-existing deficiency of the anterior cruciate ligament and severe functionally instability is a difficult subgroup to manage. There is considerable debate regarding management of young patients with isolated unicompartment osteoarthritis and concomitant ACL deficiency. A retrospective analysis of was done in 9 patients with symptomatic osteoarthritis with ACL deficiencies and functional instability that were treated with unicompartment knee arthroplasty and ACL reconstruction between April 2002 and June 2005. The average arc of flexion was 119 degrees (range 85 degrees to 135 degrees) preoperatively and 125 degrees (range 105 degrees to 140 degrees). There were no signs of instability during the follow up of patients. No patients in this group were reoperated. In this small series we have shown that instability can be corrected and pain relieved by this combined procedure. PMID- 20017918 TI - Long term outcomes of antiretroviral therapy in a large HIV/AIDS care clinic in urban South Africa: a prospective cohort study. AB - BACKGROUND: Clinical, immunologic and virologic outcomes at large HIV/AIDS care clinics in resource poor settings are poorly described beyond the first year of highly active antiretroviral treatment (HAART). We aimed to prospectively evaluate long-term treatment outcomes at a large scale HIV/AIDS care clinic in South Africa. METHODS: Cohort study of patients initiating HAART between April 1, 2004 and March 13, 2007, and followed up until April 1, 2008 at a public HIV/AIDS care clinic in Johannesburg, South Africa. We performed time to event analysis on key treatment outcomes and program impact parameters including mortality, retention in care, CD4 count gain, virologic success and first line regimen durability. RESULTS: 7583 HIV-infected patients initiated care and contributed to 161,000 person months follow up. Overall mortality rate was low (2.9 deaths per 100 person years, 95% CI 2.6-3.2), but high in the first three months of HAART (8.4 per 100 person years, 95% CI 7.2-9.9). Long-term on-site retention in care was relatively high (74.4% at 4 years, 95%CI 73.2-75.6). CD4 count was above 200 cells/mm(3 )after 6 months of treatment in almost all patients. By the fourth year of HAART, the majority (59.6%, 95%CI 57.8-61.4) of patients had at least one first line drug (mainly stavudine) substituted. Women were twice as likely to experience drug substitution (OR 1.97, 95% CI 1.80-2.16). By 6 months of HAART, 90.8% suppressed virus below 400 copies. Among those with initial viral suppression, 9.4% (95% CI 8.5-10.3%) had viral rebound within one year of viral suppression, 16.8% (95% CI 15.5-18.1) within 2 years, and 20.6% (95% CI 18.9 22.4) within 3 years of initial suppression. Only 10% of women and 13% of men initiated second line HAART. CONCLUSION: Despite advanced disease presentation and a very large-scale program, high quality care was achieved as indicated by good long-term clinical, immunologic and virologic outcomes and a low rate of second line HAART initiation. High rates of single drug substitution suggest that the public health approach to HAART could be further improved by the use of a more durable first line regimen. PMID- 20017915 TI - A new miniaturized system for extracorporeal membrane oxygenation in adult respiratory failure. AB - INTRODUCTION: Mortality of severe acute respiratory distress syndrome in adults is still unacceptably high. Extracorporeal membrane oxygenation (ECMO) could represent an important treatment option, if complications were reduced by new technical developments. METHODS: Efficiency, side effects and outcome of treatment with a new miniaturized device for veno-venous extracorporeal gas transfer were analysed in 60 consecutive patients with life-threatening respiratory failure. RESULTS: A rapid increase of partial pressure of arterial oxygen/fraction of inspired oxygen (PaO2/FiO2) from 64 (48 to 86) mmHg to 120 (84 to 171) mmHg and a decrease of PaCO2 from 63 (50 to 80) mmHg to 33 (29 to 39) mmHg were observed after start of the extracorporeal support (P < 0.001). Gas exchange capacity of the device averaged 155 (116 to 182) mL/min for oxygen and 210 (164 to 251) mL/min for carbon dioxide. Ventilatory parameters were reduced to a highly protective mode, allowing a fast reduction of tidal volume from 495 (401 to 570) mL to 336 (292 to 404) mL (P < 0.001) and of peak inspiratory pressure from 36 (32 to 40) cmH2O to 31 (28 to 35) cmH2O (P < 0.001). Transfusion requirements averaged 0.8 (0.4 to 1.8) units of red blood cells per day. Sixty two percent of patients were weaned from the extracorporeal system, and 45% survived to discharge. CONCLUSIONS: Veno-venous extracorporeal membrane oxygenation with a new miniaturized device supports gas transfer effectively, allows for highly protective ventilation and is very reliable. Modern ECMO technology extends treatment opportunities in severe lung failure. PMID- 20017916 TI - Thermogenic effect of meltdown RTD energy drink in young healthy women: a double blind, cross-over design study. AB - BACKGROUND: The purpose of this study was to examine the acute metabolic effects of a high-energy drink in healthy, physically-active women. METHODS: Ten women (20.4 +/- 0.70 y; 166.9 +/- 7.2 cm; 67.0 +/- 7.0 kg; 29.6 +/- 6.5% body fat) underwent two testing sessions administered in a randomized and double-blind fashion. Subjects reported to the laboratory in a 3-hr post-absorptive state and were provided either 140 ml of the high-energy drink (SUP; commercially marketed as Meltdown RTD) or placebo (P). Subjects consumed two 70 ml doses of SUP or P, separated by 30 min and rested in a semi-recumbent position for 3 hours. Resting oxygen consumption (VO2) and heart rate (HR) were determined every 5 min during the first 30 min and every 10 min during the next 150 min. Blood pressure (BP) was determined every 15 min during the first 30 min and every 30 min thereafter. Area under the curve (AUC) analysis was computed for VO2, whereas a 3-hour average and hourly averages were calculated for respiratory quotient (RQ), total kcal, HR, BP, and profile of mood states (POMS). RESULTS: AUC analysis revealed a 10.8% difference (p = 0.03) in VO2 between SUP and P. No difference in VO2 was seen between the groups in the first hour, but VO2 in SUP was significantly greater than P in the second (13.9%, p = 0.01) and third hours (11.9%, p = 0.03). A difference (p = 0.03) in energy expenditure was seen between SUP (1.09 +/- 0.10 kcal x min-1) and P (0.99 +/- 0.09 kcal x min-1) for the 3-hour period. Although no difference in energy expenditure was seen in the first hour, significant differences between SUP and P were observed in the second (1.10 +/- 0.11 kcal x min-1 and 0.99 +/- 0.09 kcal x min-1, respectively; p = 0.02) and third hour (1.08 +/- 0.11 kcal x min-1 and 0.99 +/- 0.09 kcal x min-1, respectively; p = 0.05). Average systolic BP was significantly higher (p = 0.007) for SUP (110.0 +/ 3.9 mmHg) compared to P (107.3 +/- 4.4 mmHg). No differences were seen in HR, diastolic BP, or POMS at any time point. CONCLUSIONS: Results showed a significant increase in energy expenditure in young, healthy women following an acute ingestion of a high-energy drink. PMID- 20017917 TI - ECRG4 is a candidate tumor suppressor gene frequently hypermethylated in colorectal carcinoma and glioma. AB - BACKGROUND: Cancer cells display widespread changes in DNA methylation that may lead to genetic instability by global hypomethylation and aberrant silencing of tumor suppressor genes by focal hypermethylation. In turn, altered DNA methylation patterns have been used to identify putative tumor suppressor genes. METHODS: In a methylation screening approach, we identified ECRG4 as a differentially methylated gene. We analyzed different cancer cells for ECRG4 promoter methylation by COBRA and bisulfite sequencing. Gene expression analysis was carried out by semi-quantitative RT-PCR. The ECRG4 coding region was cloned and transfected into colorectal carcinoma cells. Cell growth was assessed by MTT and BrdU assays. ECRG4 localization was analyzed by fluorescence microscopy and Western blotting after transfection of an ECRG4-eGFP fusion gene. RESULTS: We found a high frequency of ECRG4 promoter methylation in various cancer cell lines. Remarkably, aberrant methylation of ECRG4 was also found in primary human tumor tissues, including samples from colorectal carcinoma and from malignant gliomas. ECRG4 hypermethylation associated strongly with transcriptional silencing and its expression could be re-activated in vitro by demethylating treatment with 5-aza-2'-deoxycytidine. Overexpression of ECRG4 in colorectal carcinoma cells led to a significant decrease in cell growth. In transfected cells, ECRG4 protein was detectable within the Golgi secretion machinery as well as in the culture medium. CONCLUSIONS: ECRG4 is silenced via promoter hypermethylation in different types of human cancer cells. Its gene product may act as inhibitor of cell proliferation in colorectal carcinoma cells and may play a role as extracellular signaling molecule. PMID- 20017919 TI - Innovations on a shoestring: a study of a collaborative community-based Aboriginal mental health service model in rural Canada. AB - BACKGROUND: Collaborative, culturally safe services that integrate clinical approaches with traditional Aboriginal healing have been hailed as promising approaches to ameliorate the high rates of mental health problems in Aboriginal communities in Canada. Overcoming significant financial and human resources barriers, a mental health team in northern Ontario is beginning to realize this ideal. We studied the strategies, strengths and challenges related to collaborative Aboriginal mental health care. METHODS: A participatory action research approach was employed to evaluate the Knaw Chi Ge Win services and their place in the broader mental health system. Qualitative methods were used as the primary source of data collection and included document review, ethnographic interviews with 15 providers and 23 clients; and 3 focus groups with community workers and managers. RESULTS: The Knaw Chi Ge Win model is an innovative, community-based Aboriginal mental health care model that has led to various improvements in care in a challenging rural, high needs environment. Formal opportunities to share information, shared protocols and ongoing education support this model of collaborative care. Positive outcomes associated with this model include improved quality of care, cultural safety, and integration of traditional Aboriginal healing with clinical approaches. Ongoing challenges include chronic lack of resources, health information and the still cursory understanding of Aboriginal healing and outcomes. CONCLUSIONS: This model can serve to inform collaborative care in other rural and Indigenous mental health systems. Further research into traditional Aboriginal approaches to mental health is needed to continue advances in collaborative practice in a clinical setting. PMID- 20017921 TI - TILLING in the two-rowed barley cultivar 'Barke' reveals preferred sites of functional diversity in the gene HvHox1. AB - BACKGROUND: The economic importance of cereals such as barley, and the demand for improved yield and quality require a better understanding of the genetic components that modulate biologically and commercially relevant traits. While Arabidopsis thaliana is the premiere model plant system, the spectrum of its traits cannot address all of the fundamental questions of crop plant development. Unlike Arabidopsis, barley is both a crop and a model system for scientific research, and it is increasingly being used for genetic and molecular investigations into the conserved biological processes of cereals. A common challenge in genetic studies in plants with large genomes arises from the very time-consuming work of associating mutant phenotypes with gene sequence information, especially if insertion mutagenesis is not routine, as in barley. Reverse genetics based on chemical mutagenesis represents the best solution to this obstacle. FINDINGS: In barley, we generated a new TILLING (Targeting Local Lesions IN Genomes) resource comprising 10,279 M(2 )mutants in the two-rowed malting cultivar 'Barke,' which has been used in the generation of other genomic resources in barley (~150,000 ESTs, DH mapping population). The value of this new resource was tested using selected candidate genes. An average frequency of approximately one mutation per 0.5 Mb was determined by screening ten fragments of six different genes. The ethyl methanesulphonate (EMS)mutagenesis efficiency was studied by recording and relating the mutagenesis-dependent effects found in the three mutant generations (M(1)-M(3)). A detailed analysis was performed for the homeodomain-leucine-zipper (HD-ZIP) gene HvHox1. Thirty-one mutations were identified by screening a 1,270-bp fragment in 7,348 M(2 )lines. Three of the newly identified mutants exhibited either a six-rowed or an intermedium-spike phenotype, and one mutant displayed a significantly altered spikelet morphology compared to that of the 'Barke' wild type. Our results indicate a bias in the frequency of independent functional mutations at specific base pair (bp) positions within the gene HvHox1. CONCLUSIONS: A new TILLING population was developed as a resource for high-throughput gene discovery in an alternative barley germplasm. Pilot screening demonstrated a similar or even slightly higher mutation frequency when compared to previously published barley TILLING populations that should allow for the identification of diverse allelic variation. Partial phenotypic evaluation of the M(2 )and M(3 )generations has revealed the presence of a wide spectrum of morphological diversity that highlights the great potential of this resource for use in forward genetic screens. Altogether, our study shows the efficiency of screening and the applicability of the new TILLING population for genetic studies in the barley crop model system. PMID- 20017920 TI - Plastic or metal stents for benign extrahepatic biliary strictures: a systematic review. AB - BACKGROUND: Benign biliary strictures may be a consequence of surgical procedures, chronic pancreatitis or iatrogenic injuries to the ampulla. Stents are increasingly being used for this indication, however it is not completely clear which stent type should be preferred. METHODS: A systematic review on stent placement for benign extrahepatic biliary strictures was performed after searching PubMed and EMBASE databases. Data were pooled and evaluated for technical success, clinical success and complications. RESULTS: In total, 47 studies (1116 patients) on outcome of stent placement were identified. No randomized controlled trials (RCTs), one non-randomized comparative studies and 46 case series were found. Technical success was 98,9% for uncovered self expandable metal stents (uSEMS), 94,8% for single plastic stents and 94,0% for multiple plastic stents. Overall clinical success rate was highest for placement of multiple plastic stents (94,3%) followed by uSEMS (79,5%) and single plastic stents (59.6%). Complications occurred more frequently with uSEMS (39.5%) compared with single plastic stents (36.0%) and multiple plastic stents (20,3%). CONCLUSION: Based on clinical success and risk of complications, placement of multiple plastic stents is currently the best choice. The evolving role of cSEMS placement as a more patient friendly and cost effective treatment for benign biliary strictures needs further elucidation. There is a need for RCTs comparing different stent types for this indication. PMID- 20017923 TI - Identification of the mechanisms that drive the toxicity of TiO(2 )particulates: the contribution of physicochemical characteristics. AB - This review focuses on outlining the toxicity of titanium dioxide (TiO(2)) particulates in vitro and in vivo, in order to understand their ability to detrimentally impact on human health. Evaluating the hazards associated with TiO(2 )particles is vital as it enables risk assessments to be conducted, by combining this information with knowledge on the likely exposure levels of humans. This review has concentrated on the toxicity of TiO(2), due to the fact that the greatest number of studies by far have evaluated the toxicity of TiO(2), in comparison to other metal oxide particulates. This derives from historical reasons (whereby the size dependency of particulate toxicity was first realised for TiO(2)) and due to its widespread application within consumer products (such as sunscreens). The pulmonary and dermal hazards of TiO(2 )have been a particular focus of the available studies, due to the past use of TiO(2 )as a (negative) control when assessing the pulmonary toxicity of particulates, and due to its incorporation within consumer products such as sunscreens. Mechanistic processes that are critical to TiO(2 )particulate toxicity will also be discussed and it is apparent that, in the main, the oxidant driven inflammatory, genotoxic and cytotoxic consequences associated with TiO(2 )exposure, are inherently linked, and are evident both in vivo and in vitro. The attributes of TiO(2 )that have been identified as being most likely to drive the observed toxicity include particle size (and therefore surface area), crystallinity (and photocatalytic activity), surface chemistry, and particle aggregation/agglomeration tendency. The experimental set up also influences toxicological outcomes, so that the species (or model) used, route of exposure, experiment duration, particle concentration and light conditions are all able to influence the findings of investigations. In addition, the applicability of the observed findings for particular TiO(2 )forms, to TiO(2 )particulates in general, requires consideration. At this time it is inappropriate to consider the findings for one TiO(2 )form as being representative for TiO(2 )particulates as a whole, due to the vast number of available TiO(2 )particulate forms and large variety of potential tissue and cell targets that may be affected by exposure. Thus emphasising that the physicochemical characteristics are fundamental to their toxicity. PMID- 20017922 TI - Making stillbirths count, making numbers talk - issues in data collection for stillbirths. AB - BACKGROUND: Stillbirths need to count. They constitute the majority of the world's perinatal deaths and yet, they are largely invisible. Simply counting stillbirths is only the first step in analysis and prevention. From a public health perspective, there is a need for information on timing and circumstances of death, associated conditions and underlying causes, and availability and quality of care. This information will guide efforts to prevent stillbirths and improve quality of care. DISCUSSION: In this report, we assess how different definitions and limits in registration affect data capture, and we discuss the specific challenges of stillbirth registration, with emphasis on implementation. We identify what data need to be captured, we suggest a dataset to cover core needs in registration and analysis of the different categories of stillbirths with causes and quality indicators, and we illustrate the experience in stillbirth registration from different cultural settings. Finally, we point out gaps that need attention in the International Classification of Diseases and review the qualities of alternative systems that have been tested in low- and middle-income settings. SUMMARY: Obtaining high-quality data will require consistent definitions for stillbirths, systematic population-based registration, better tools for surveys and verbal autopsies, capacity building and training in procedures to identify causes of death, locally adapted quality indicators, improved classification systems, and effective registration and reporting systems. PMID- 20017924 TI - Evidence for a novel coding sequence overlapping the 5'-terminal approximately 90 codons of the gill-associated and yellow head okavirus envelope glycoprotein gene. AB - The genus Okavirus (order Nidovirales) includes a number of viruses that infect crustaceans, causing major losses in the shrimp industry. These viruses have a linear positive-sense ssRNA genome of approximately 26-27 kb, encoding a large replicase polyprotein that is expressed from the genomic RNA, and several additional proteins that are expressed from a nested set of 3'-coterminal subgenomic RNAs. In this brief report, we describe the bioinformatic discovery of a new, apparently coding, ORF that overlaps the 5' end of the envelope glycoprotein encoding sequence, ORF3, in the +2 reading frame. The new ORF has a strong coding signature and, in fact, is more conserved at the amino acid level than the overlapping region of ORF3. We propose that translation of the new ORF initiates at a conserved AUG codon separated by just 2 nt from the ORF3 AUG initiation codon, resulting in a novel 86 amino acid protein. PMID- 20017925 TI - Cultured skin microbiota attracts malaria mosquitoes. AB - BACKGROUND: Host-seeking of the African malaria mosquito, Anopheles gambiae sensu stricto, is guided by human odours. The precise nature of the odours, and the composition of attractive blends of volatiles, remains largely unknown. Skin microbiota plays an important role in the production of human body odours. It is hypothesized that host attractiveness and selection of An. gambiae is affected by the species composition, density, and metabolic activity of the skin microbiota. A study is presented in which the production and constituency of volatile organic compounds (VOCs) by human skin microbiota is examined and the behavioural responses of An. gambiae to VOCs from skin microbiota are investigated. METHODS: Blood agar plates incubated with skin microbiota from human feet or with a reference strain of Staphylococcus epidermidis were tested for their attractiveness to An. gambiae in olfactometer bioassays and indoor trapping experiments. Entrained air collected from blood agar plates incubated with natural skin microbiota or with S. epidermidis were analysed using GC-MS. A synthetic blend of the compounds identified was tested for its attractiveness to An. gambiae. Behavioural data were analysed by a chi(2)-test and GLM. GC-MS results were analysed by fitting an exponential regression line to test the effect of the concentration of bacteria. RESULTS: More An. gambiae were caught with blood agar plates incubated with skin bacteria than with sterile blood agar plates, with a significant effect of incubation time and dilution of the skin microbiota. When bacteria from the feet of four other volunteers were tested, similar effects were found. Fourteen putative attractants were found in the headspace of the skin bacteria. A synthetic blend of 10 of these was attractive to An. gambiae. CONCLUSIONS: The discovery that volatiles produced by human skin microorganisms in vitro mediate An. gambiae host-seeking behaviour creates new opportunities for the development of odour-baited trapping systems. Additionally, identification of bacterial volatiles provides a new method to develop synthetic blends, attractive to An. gambiae and possibly other anthropophilic disease vectors. PMID- 20017926 TI - The complete genome sequence of Xanthomonas albilineans provides new insights into the reductive genome evolution of the xylem-limited Xanthomonadaceae. AB - BACKGROUND: The Xanthomonadaceae family contains two xylem-limited plant pathogenic bacterial species, Xanthomonas albilineans and Xylella fastidiosa. X. fastidiosa was the first completely sequenced plant pathogen. It is insect vectored, has a reduced genome and does not possess hrp genes which encode a Type III secretion system found in most plant pathogenic bacteria. X. fastidiosa was excluded from the Xanthomonas group based on phylogenetic analyses with rRNA sequences. RESULTS: The complete genome of X. albilineans was sequenced and annotated. X. albilineans, which is not known to be insect-vectored, also has a reduced genome and does not possess hrp genes. Phylogenetic analysis using X. albilineans genomic sequences showed that X. fastidiosa belongs to the Xanthomonas group. Order of divergence of the Xanthomonadaceae revealed that X. albilineans and X. fastidiosa experienced a convergent reductive genome evolution during their descent from the progenitor of the Xanthomonas genus. Reductive genome evolutions of the two xylem-limited Xanthomonadaceae were compared in light of their genome characteristics and those of obligate animal symbionts and pathogens. CONCLUSION: The two xylem-limited Xanthomonadaceae, during their descent from a common ancestral parent, experienced a convergent reductive genome evolution. Adaptation to the nutrient-poor xylem elements and to the cloistered environmental niche of xylem vessels probably favoured this convergent evolution. However, genome characteristics of X. albilineans differ from those of X. fastidiosa and obligate animal symbionts and pathogens, indicating that a distinctive process was responsible for the reductive genome evolution in this pathogen. The possible role in genome reduction of the unique toxin albicidin, produced by X. albilineans, is discussed. PMID- 20017927 TI - Analysis of the stoichiometric metal activation of methionine aminopeptidase. AB - BACKGROUND: Methionine aminopeptidase (MetAP) is a ubiquitous enzyme required for cell survival and an attractive target for antibacterial and anticancer drug development. The number of a divalent metal required for catalysis is under intense debate. E. coli MetAP was shown to be fully active with one equivalent of metal by graphical analysis, but it was inferred to require at least two metals by a Hill equation model. Herein, we report a mathematical model and detailed analysis of the stoichiometric activation of MetAP by metal cofactors. RESULTS: Because of diverging results with significant implications in drug discovery, the experimental titration curve for Co2+ activating MetAP was analyzed by fitting with a multiple independent binding sites (MIBS) model, and the quality of the fitting was compared to that of the Hill equation. The fitting by the MIBS model was clearly superior and indicated that complete activity is observed at a one metal to one protein ratio. The shape of the titration curve was also examined for activation of metalloenzymes in general by one or two metals. CONCLUSIONS: Considering different scenarios of MetAP activation by one or two metal ions, it is concluded that E. coli MetAP is fully active as a monometalated enzyme. Our approach can be of value in proper determination of the number of cations needed for catalysis by metalloenzymes. PMID- 20017928 TI - Photodynamic therapy in the therapy for recurrent/persistent nasopharyngeal cancer. AB - To determine the efficacy of Photodynamic therapy of patients with recurrent Nasopharyngeal Carcinoma we reviewed all available literature. Since the treatment options for recurrent or persistent Nasopharyngeal Carcinoma are limited, the survival rates poor and the complications severe; there is definitely a place for alternative treatment modalities with more efficacy and less morbidity. Photodynamic therapy (PDT) has the potential to be a very effective local treatment modality for recurrent or persistent nasopharyngeal cancer, without the severe side effects seen with re-irradiation. This review shows all reported results of Photodynamic therapy in the treatment for Nasopharyngeal Carcinoma. PMID- 20017929 TI - Eukaryotic large nucleo-cytoplasmic DNA viruses: clusters of orthologous genes and reconstruction of viral genome evolution. AB - BACKGROUND: The Nucleo-Cytoplasmic Large DNA Viruses (NCLDV) comprise an apparently monophyletic class of viruses that infect a broad variety of eukaryotic hosts. Recent progress in isolation of new viruses and genome sequencing resulted in a substantial expansion of the NCLDV diversity, resulting in additional opportunities for comparative genomic analysis, and a demand for a comprehensive classification of viral genes. RESULTS: A comprehensive comparison of the protein sequences encoded in the genomes of 45 NCLDV belonging to 6 families was performed in order to delineate cluster of orthologous viral genes. Using previously developed computational methods for orthology identification, 1445 Nucleo-Cytoplasmic Virus Orthologous Groups (NCVOGs) were identified of which 177 are represented in more than one NCLDV family. The NCVOGs were manually curated and annotated and can be used as a computational platform for functional annotation and evolutionary analysis of new NCLDV genomes. A maximum-likelihood reconstruction of the NCLDV evolution yielded a set of 47 conserved genes that were probably present in the genome of the common ancestor of this class of eukaryotic viruses. This reconstructed ancestral gene set is robust to the parameters of the reconstruction procedure and so is likely to accurately reflect the gene core of the ancestral NCLDV, indicating that this virus encoded a complex machinery of replication, expression and morphogenesis that made it relatively independent from host cell functions. CONCLUSIONS: The NCVOGs are a flexible and expandable platform for genome analysis and functional annotation of newly characterized NCLDV. Evolutionary reconstructions employing NCVOGs point to complex ancestral viruses. PMID- 20017930 TI - Implementing a fax referral program for quitline smoking cessation services in urban health centers: a qualitative study. AB - BACKGROUND: Fax referral services that connect smokers to state quitlines have been implemented in 49 U.S. states and territories and promoted as a simple solution to improving smoker assistance in medical practice. This study is an in depth examination of the systems-level changes needed to implement and sustain a fax referral program in primary care. METHODS: The study involved implementation of a fax referral system paired with a chart stamp prompting providers to identify smoking patients, provide advice to quit and refer interested smokers to a state-based fax quitline. Three focus groups (n = 26) and eight key informant interviews were conducted with staff and physicians at two clinics after the intervention. We used the Chronic Care Model as a framework to analyze the data, examining how well the systems changes were implemented and the impact of these changes on care processes, and to develop recommendations for improvement. RESULTS: Physicians and staff described numerous benefits of the fax referral program for providers and patients but pointed out significant barriers to full implementation, including the time-consuming process of referring patients to the Quitline, substantial patient resistance, and limitations in information and care delivery systems for referring and tracking smokers. Respondents identified several strategies for improving integration, including simplification of the referral form, enhanced teamwork, formal assignment of responsibility for referrals, ongoing staff training and patient education. Improvements in Quitline feedback were needed to compensate for clinics' limited internal information systems for tracking smokers. CONCLUSIONS: Establishing sustainable linkages to quitline services in clinical sites requires knowledge of existing patterns of care and tailored organizational changes to ensure new systems are prioritized, easily integrated into current office routines, formally assigned to specific staff members, and supported by internal systems that ensure adequate tracking and follow up of smokers. Ongoing staff training and patient self-management techniques are also needed to ease the introduction of new programs and increase their acceptability to smokers. PMID- 20017931 TI - The structure of pyogenecin immunity protein, a novel bacteriocin-like immunity protein from Streptococcus pyogenes. AB - BACKGROUND: Many Gram-positive lactic acid bacteria (LAB) produce anti-bacterial peptides and small proteins called bacteriocins, which enable them to compete against other bacteria in the environment. These peptides fall structurally into three different classes, I, II, III, with class IIa being pediocin-like single entities and class IIb being two-peptide bacteriocins. Self-protective cognate immunity proteins are usually co-transcribed with these toxins. Several examples of cognates for IIa have already been solved structurally. Streptococcus pyogenes, closely related to LAB, is one of the most common human pathogens, so knowledge of how it competes against other LAB species is likely to prove invaluable. RESULTS: We have solved the crystal structure of the gene-product of locus Spy_2152 from S. pyogenes, (PDB:2fu2), and found it to comprise an anti parallel four-helix bundle that is structurally similar to other bacteriocin immunity proteins. Sequence analyses indicate this protein to be a possible immunity protein protective against class IIa or IIb bacteriocins. However, given that S. pyogenes appears to lack any IIa pediocin-like proteins but does possess class IIb bacteriocins, we suggest this protein confers immunity to IIb-like peptides. CONCLUSIONS: Combined structural, genomic and proteomic analyses have allowed the identification and in silico characterization of a new putative immunity protein from S. pyogenes, possibly the first structure of an immunity protein protective against potential class IIb two-peptide bacteriocins. We have named the two pairs of putative bacteriocins found in S. pyogenes pyogenecin 1, 2, 3 and 4. PMID- 20017933 TI - Remaining life expectancy among older people in a rural area of Vietnam: trends and socioeconomic inequalities during a period of multiple transitions. AB - BACKGROUND: Better understanding of the trends and disparities in health at old age in terms of life expectancy will help to provide appropriate responses to the growing needs of health and social care for the older population in the context of limited resources. As a result of rapid economic, demographic and epidemiological changes, the number of people aged 60 and over in Vietnam is increasing rapidly, from 6.7% in 1979 to 9.2% in 2006. Life expectancy at birth has increased but not much are known about changes in old ages. This study assesses the trends and socioeconomic inequalities in RLE at age 60 in a rural area in an effort to highlight this vulnerable group and to anticipate their future health and social needs. METHODS: An abridged life table adjusted for small area data was used to estimate cohort life expectancies at old age and the corresponding 95% confidence intervals from longitudinal data collected by FilaBavi DSS during 1999-2006, which covered 7,668 people at age 60+ with 43,272 person-years, out of a total of 64,053 people with 388,278 person-years. Differences in life expectancy were examined according to socioeconomic factors, including socio-demographic characteristics, wealth, poverty and living arrangements. RESULTS: Life expectancies at age 60 have increased by approximately one year from the period 1999-2002 to 2003-2006. The increases are observed in both sexes, but are significant among females and relate to improvements among those who belong to the middle and upper household wealth quintiles. However, life expectancy tends to decrease in the most vulnerable groups. There is a wide gap in life expectancy according to poverty status and living arrangements, and the gap by poverty status has widened over the study period. The gender gap in life expectancy is consistent across all socioeconomic groups and tends to be wider amongst the more disadvantaged population. CONCLUSIONS: There is a trend of increasing life expectancy among older people in rural areas of Vietnam. Inequalities in life expectancy exist between socioeconomic groups, especially between different poverty levels and also patterns of living arrangements. These inequalities should be addressed by appropriate social and health policies with stronger targeting of the poorest and most disadvantaged groups. PMID- 20017932 TI - Adolescent type 1 Diabetes Cardio-renal Intervention Trial (AdDIT). AB - BACKGROUND: The prognosis for young people diagnosed with diabetes during childhood remains poor and this is mainly related to the long-term risk of developing vascular complications.Microalbuminuria identifies subjects at risk for diabetic nephropathy (DN) and cardiovascular disease (CVD). It is often detected in adolescence but is rarely treated before the age of 18 years, as at the end of puberty albumin excretion may decline and in some subjects will return into the normal range. However, evidence indicates that subjects with both transient and persistent microalbuminuria have experienced renal damage during puberty and thus reno-protection to prevent long-term complications is warranted. In adults with diabetes and microalbuminuria, the use of angiotensin converting enzyme inhibitors (ACEI) and Statins is increasing, and in order to determine whether these agents are of value in the adolescent population a large randomized controlled clinical trial is needed. METHODS/DESIGN: The Adolescent type 1 Diabetes cardio-renal Intervention Trial (AdDIT) is a multi-center, randomized, double-blind, placebo-controlled trial of ACEI and Statin therapy in adolescents with type 1 diabetes. 500 high-risk adolescents, defined on the basis of their albumin excretion, are randomized to receive either ACEI (Quinapril) or Statins (Atorvastatin) or combination therapy or placebo for 3-4 years. There will also be a parallel open observational study, based on the follow-up of 400 low-risk non-randomized adolescents. The major endpoint of the study is the change in albumin excretion; secondary endpoints include markers of CVD, renal function, retinopathy, quality of life combined with assessment of compliance and potential health economic benefits. DISCUSSION: AdDIT will provide important data on the potential renal and cardiovascular protective effects of ACEI and Statins in high risk adolescents. Long-term follow-up of the randomized subjects will provide direct evidence of disease outcomes, in addition to the data on early surrogate measures of DN and CVD. Follow-up of non-randomized low-risk subjects will determine the potential impact of intervention on DN and CVD. AdDIT will help to determine whether, in addition to encouraging young people to achieve good glycaemic control, pharmacological cardio-renal protection should also be implemented. EUDRACT NUMBER: 2007-001039-72 TRIAL REGISTRATION NUMBER: ISRCTN91419926. PMID- 20017934 TI - Argon: neuroprotection in in vitro models of cerebral ischemia and traumatic brain injury. AB - INTRODUCTION: Recently, it has been shown in several experimental settings that the noble gases xenon and helium have neuroprotective properties. In this study we tested the hypothesis that the noble gas argon has a neuroprotective potential as well. Since traumatic brain injury and stroke are widespread and generate an enormous economic and social burden, we investigated the possible neuroprotective effect in in vitro models of traumatic brain injury and cerebral ischemia. METHODS: Organotypic hippocampal slice cultures from mice pups were subjected to either oxygen-glucose deprivation or to a focal mechanical trauma and subsequently treated with three different concentrations (25, 50 and 74%) of argon immediately after trauma or with a two-or-three-hour delay. After 72 hours of incubation tissue injury assessment was performed using propidium iodide, a staining agent that becomes fluorescent when it diffuses into damaged cells via disintegrated cell membranes. RESULTS: We could show argon's neuroprotective effects at different concentrations when applied directly after oxygen-glucose deprivation or trauma. Even three hours after application, argon was still neuroprotective. CONCLUSIONS: Argon showed a neuroprotective effect in both in vitro models of oxygen-glucose deprivation and traumatic brain injury. Our promising results justify further in vivo animal research. PMID- 20017935 TI - Effect of auditory feedback differs according to side of hemiparesis: a comparative pilot study. AB - BACKGROUND: Following stroke, patients frequently demonstrate loss of motor control and function and altered kinematic parameters of reaching movements. Feedback is an essential component of rehabilitation and auditory feedback of kinematic parameters may be a useful tool for rehabilitation of reaching movements at the impairment level. The aim of this study was to investigate the effect of 2 types of auditory feedback on the kinematics of reaching movements in hemiparetic stroke patients and to compare differences between patients with right (RHD) and left hemisphere damage (LHD). METHODS: 10 healthy controls, 8 stroke patients with LHD and 8 with RHD were included. Patient groups had similar levels of upper limb function. Two types of auditory feedback (spatial and simple) were developed and provided online during reaching movements to 9 targets in the workspace. Kinematics of the upper limb were recorded with an electromagnetic system. Kinematics were compared between groups (Mann Whitney test) and the effect of auditory feedback on kinematics was tested within each patient group (Friedman test). RESULTS: In the patient groups, peak hand velocity was lower, the number of velocity peaks was higher and movements were more curved than in the healthy group. Despite having a similar clinical level, kinematics differed between LHD and RHD groups. Peak velocity was similar but LHD patients had fewer velocity peaks and less curved movements than RHD patients. The addition of auditory feedback improved the curvature index in patients with RHD and deteriorated peak velocity, the number of velocity peaks and curvature index in LHD patients. No difference between types of feedback was found in either patient group. CONCLUSION: In stroke patients, side of lesion should be considered when examining arm reaching kinematics. Further studies are necessary to evaluate differences in responses to auditory feedback between patients with lesions in opposite cerebral hemispheres. PMID- 20017936 TI - Molecular and biochemical characterization of urease and survival of Yersinia enterocolitica biovar 1A in acidic pH in vitro. AB - BACKGROUND: Yersinia enterocolitica, an important food- and water-borne enteric pathogen is represented by six biovars viz. 1A, 1B, 2, 3, 4 and 5. Despite the lack of recognized virulence determinants, some biovar 1A strains have been reported to produce disease symptoms resembling that produced by known pathogenic biovars (1B, 2-5). It is therefore imperative to identify determinants that might contribute to the pathogenicity of Y. enterocolitica biovar 1A strains. Y. enterocolitica invariably produces urease and the role of this enzyme in the virulence of biovar 1B and biovar 4 strains has been reported recently. The objective of this work was to study genetic organization of the urease (ure) gene complex of Y. enterocolitica biovar 1A, biochemical characterization of the urease, and the survival of these strains under acidic conditions in vitro. RESULTS: The ure gene complex (ureABCEFGD) of Y. enterocolitica biovar 1A included three structural and four accessory genes, which were contiguous and was flanked by a urea transport (yut) gene on the 3' side. Differences were identified in ure gene complex of biovar 1A strain compared to biovar 1B and 4 strains. This included a smaller ureB gene and larger intergenic regions between the structural genes. The crude urease preparation exhibited optimal pH and temperature of 5.5 and 65 degrees C respectively, and Michaelis-Menten kinetics with a Km of 1.7 +/- 0.4 mM urea and Vmax of 7.29 +/- 0.42 micromol of ammonia released/min/mg protein. The urease activity was dependent on growth temperature and growth phase of Y. enterocolitica biovar 1A, and the presence of nickel in the medium. The molecular mass of the enzyme was > 545 kDa and an isoelectric point of 5.2. The number of viable Y. enterocolitica biovar 1A decreased significantly when incubated at pH 2.5 for 2 h. However, no such decrease was observed at this pH in the presence of urea. CONCLUSIONS: The ure gene cluster of biovar 1A strains though similar to biovar 1B and 4 strains, exhibited important differences. The study also showed the ability of biovar 1A strains of Y. enterocolitica to survive at highly acidic pH in vitro in the presence of urea. PMID- 20017937 TI - Primary ciliogenesis defects are associated with human astrocytoma/glioblastoma cells. AB - BACKGROUND: Primary cilia are non-motile sensory cytoplasmic organelles that have been implicated in signal transduction, cell to cell communication, left and right pattern embryonic development, sensation of fluid flow, regulation of calcium levels, mechanosensation, growth factor signaling and cell cycle progression. Defects in the formation and/or function of these structures underlie a variety of human diseases such as Alstrom, Bardet-Biedl, Joubert, Meckel-Gruber and oral-facial-digital type 1 syndromes. The expression and function of primary cilia in cancer cells has now become a focus of attention but has not been studied in astrocytomas/glioblastomas. To begin to address this issue, we compared the structure and expression of primary cilia in a normal human astrocyte cell line with five human astrocytoma/glioblastoma cell lines. METHODS: Cultured normal human astrocytes and five human astrocytoma/glioblastoma cell lines were examined for primary cilia expression and structure using indirect immunofluorescence and electron microscopy. Monospecific antibodies were used to detect primary cilia and map the relationship between the primary cilia region and sites of endocytosis. RESULTS: We show that expression of primary cilia in normal astrocytes is cell cycle related and the primary cilium extends through the cell within a unique structure which we show to be a site of endocytosis. Importantly, we document that in each of the five astrocytoma/glioblastoma cell lines fully formed primary cilia are either expressed at a very low level, are completely absent or have aberrant forms, due to incomplete ciliogenesis. CONCLUSIONS: The recent discovery of the importance of primary cilia in a variety of cell functions raises the possibility that this structure may have a role in a variety of cancers. Our finding that the formation of the primary cilium is disrupted in cells derived from astrocytoma/glioblastoma tumors provides the first evidence that altered primary cilium expression and function may be part of some malignant phenotypes. Further, we provide the first evidence that ciliogenesis is not an all or none process; rather defects can arrest this process at various points, particularly at the stage subsequent to basal body association with the plasma membrane. PMID- 20017938 TI - Doctor-patient communication with people with intellectual disability--a qualitative study. AB - BACKGROUND: People with intellectual disability (ID) expressed dissatisfaction with doctor-patient communication and mentioned certain preferences for this communication (our research). Since many people with ID in the Netherlands have recently moved from residential care facilities to supported accommodations in the community, medical care for them was transferred from ID physicians (IDPs) to general practitioners (GPs) in the vicinity of the new accommodation. We addressed the following research question: 'What are the similarities and differences between the communication preferences of people with ID and the professional criteria for doctor-patient communication by GPs?' METHODS: A focus group meeting and interviews were used to identify the preferences of 12 persons with ID for good communication with their GP; these were compared with communication criteria used to assess trainee GPs, as described in the MAAS Global manual. RESULTS: Eight preferences for doctor-patient communication were formulated by the people with ID. Six of them matched the criteria used for GPs. Improvements are required as regards the time available for consultation, demonstrating physical examinations before applying them and triadic communication. CONCLUSIONS: People with ID hold strong views on communication with their doctors during consultations. GPs, people with ID and their support workers can further fine-tune their communication skills. PMID- 20017939 TI - Effects of intensive arm training with the rehabilitation robot ARMin II in chronic stroke patients: four single-cases. AB - BACKGROUND: Robot-assisted therapy offers a promising approach to neurorehabilitation, particularly for severely to moderately impaired stroke patients. The objective of this study was to investigate the effects of intensive arm training on motor performance in four chronic stroke patients using the robot ARMin II. METHODS: ARMin II is an exoskeleton robot with six degrees of freedom (DOF) moving shoulder, elbow and wrist joints. Four volunteers with chronic (>or= 12 months post-stroke) left side hemi-paresis and different levels of motor severity were enrolled in the study. They received robot-assisted therapy over a period of eight weeks, three to four therapy sessions per week, each session of one hour.Patients 1 and 4 had four one-hour training sessions per week and patients 2 and 3 had three one-hour training sessions per week. Primary outcome variable was the Fugl-Meyer Score of the upper extremity Assessment (FMA), secondary outcomes were the Wolf Motor Function Test (WMFT), the Catherine Bergego Scale (CBS), the Maximal Voluntary Torques (MVTs) and a questionnaire about ADL-tasks, progress, changes, motivation etc. RESULTS: Three out of four patients showed significant improvements (p < 0.05) in the main outcome. The improvements in the FMA scores were aligned with the objective results of MVTs. Most improvements were maintained or even increased from discharge to the six month follow-up. CONCLUSION: Data clearly indicate that intensive arm therapy with the robot ARMin II can significantly improve motor function of the paretic arm in some stroke patients, even those in a chronic state. The findings of the study provide a basis for a subsequent controlled randomized clinical trial. PMID- 20017940 TI - Chloroquine reduces arylsulphatase B activity and increases chondroitin-4 sulphate: implications for mechanisms of action and resistance. AB - BACKGROUND: The receptors for adhesion of Plasmodium falciparum-infected red blood cells (RBC) in the placenta have been identified as chondroitin-4-sulphate (C4S) proteoglycans, and the more sulphate-rich chondroitin oligosaccharides have been reported to inhibit adhesion. Since the anti-malarial drug chloroquine accumulates in lysosomes and alters normal lysosomal processes, the effects of chloroquine on the lysosomal enzyme arylsulphatase B (ASB, N-acetylgalactosamine 4-sulphatase), which removes 4-sulphate groups from chondroitin-4-sulphate, were addressed. The underlying hypothesis derived from the recognized impairment of attachment of parasite-infected erythrocytes in the placenta, when chondroitin-4 sulphation was increased. If chloroquine reduced ASB activity, leading to increased chondroitin-4-sulphation, it was hypothesized that the anti-malarial mechanism of chloroquine might derive, at least in part, from suppression of ASB. METHODS: Experimental methods involved cell culture of human placental, bronchial epithelial, and cerebrovascular cells, and the in vitro exposure of the cells to chloroquine at increasing concentrations and durations. Measurements of arylsulphatase B enzymatic activity, total sulphated glycosaminoglycans (sGAG), and chondroitin-4-sulphate (C4S) were performed using in vitro assays, following exposure to chloroquine and in untreated cell preparations. Fluorescent immunostaining of ASB was performed to determine the effect of chloroquine on cellular ASB content and localization. Mass spectrometry and high performance liquid chromatography were performed to document and to quantify the changes in chondroitin disaccharides following chloroquine exposure. RESULTS: In the human placental, bronchial epithelial, and cerebrovascular cells, exposure to increasing concentrations of chloroquine was associated with reduced ASB activity and with increased concentrations of sGAG, largely attributable to increased C4S. The study data demonstrated: 1) decline in ASB activity following chloroquine exposure; 2) inverse correlation between ASB activity and C4S content; 3) increased content of chondroitin-4-sulphate disaccharides following chloroquine exposure; and 4) decline in extent of chloroquine-induced ASB reduction with lower baseline ASB activity. Confocal microscopy demonstrated the presence of ASB along the cell periphery, indicating extra-lysosomal localization. CONCLUSIONS: The study data indicate that the therapeutic mechanism of chloroquine action may be attributable, at least in part, to reduction of ASB activity, leading to increased chondroitin-4-sulphation in human placental, bronchial epithelial, and cerebrovascular cells. In vivo, increased chondroitin-4-sulphation may reduce the attachment of P. falciparum-infected erythrocytes to human cells. Extra-lysosomal localization of ASB and reduced impact of chloroquine when baseline ASB activity is less suggest possible mechanisms of resistance to the effects of chloroquine. PMID- 20017941 TI - A taxonomy of epithelial human cancer and their metastases. AB - BACKGROUND: Microarray technology has allowed to molecularly characterize many different cancer sites. This technology has the potential to individualize therapy and to discover new drug targets. However, due to technological differences and issues in standardized sample collection no study has evaluated the molecular profile of epithelial human cancer in a large number of samples and tissues. Additionally, it has not yet been extensively investigated whether metastases resemble their tissue of origin or tissue of destination. METHODS: We studied the expression profiles of a series of 1566 primary and 178 metastases by unsupervised hierarchical clustering. The clustering profile was subsequently investigated and correlated with clinico-pathological data. Statistical enrichment of clinico-pathological annotations of groups of samples was investigated using Fisher exact test. Gene set enrichment analysis (GSEA) and DAVID functional enrichment analysis were used to investigate the molecular pathways. Kaplan-Meier survival analysis and log-rank tests were used to investigate prognostic significance of gene signatures. RESULTS: Large clusters corresponding to breast, gastrointestinal, ovarian and kidney primary tissues emerged from the data. Chromophobe renal cell carcinoma clustered together with follicular differentiated thyroid carcinoma, which supports recent morphological descriptions of thyroid follicular carcinoma-like tumors in the kidney and suggests that they represent a subtype of chromophobe carcinoma. We also found an expression signature identifying primary tumors of squamous cell histology in multiple tissues. Next, a subset of ovarian tumors enriched with endometrioid histology clustered together with endometrium tumors, confirming that they share their etiopathogenesis, which strongly differs from serous ovarian tumors. In addition, the clustering of colon and breast tumors correlated with clinico pathological characteristics. Moreover, a signature was developed based on our unsupervised clustering of breast tumors and this was predictive for disease specific survival in three independent studies. Next, the metastases from ovarian, breast, lung and vulva cluster with their tissue of origin while metastases from colon showed a bimodal distribution. A significant part clusters with tissue of origin while the remaining tumors cluster with the tissue of destination. CONCLUSION: Our molecular taxonomy of epithelial human cancer indicates surprising correlations over tissues. This may have a significant impact on the classification of many cancer sites and may guide pathologists, both in research and daily practice. Moreover, these results based on unsupervised analysis yielded a signature predictive of clinical outcome in breast cancer. Additionally, we hypothesize that metastases from gastrointestinal origin either remember their tissue of origin or adapt to the tissue of destination. More specifically, colon metastases in the liver show strong evidence for such a bimodal tissue specific profile. PMID- 20017942 TI - T-cell activation promotes tumorigenesis in inflammation-associated cancer. AB - Chronic inflammation has long been associated with a wide range of malignancies, is now widely accepted as a risk factor for development of cancer, and has been implicated as a promoter of a variety of cancers including hematopoietic malignancies. We have described a mouse model uniquely suited to examine the link between inflammation and lymphoma in which the Tax oncogene, expressed in activated T and NK cells, perpetuates chronic inflammation that begins as microscopic intraepithelial lesions and develops into inflammatory nodules, subcutaneous tumors, and large granular lymphocytic leukemia. The use of bioluminescent imaging in these mice has expanded our ability to interrogate aspects of inflammation and tumorigenesis non-invasively. Here we demonstrate that bioluminescence induction in these mice correlated with inflammation resulting from wounding, T cell activation, and exposure to chemical agents. In experiments in which long-term effects of inflammation on disease outcome were monitored, the development of lymphoma was promoted by an inflammatory stimulus. Finally we demonstrated that activation of T-cells in T-cell receptor (TCR) transgenic TAX-LUC animals dramatically exacerbated the development of subcutaneous TCR- CD16+ LGL tumors. The role of activated T-cells and acquired immunity in inflammation-associated cancers is broadly applicable to hematopoietic malignancies, and we propose these mice will be of use in dissecting mechanisms by which activated T-cells promote lymphomagenesis in vivo. PMID- 20017943 TI - Comparison of the effects of vitamin D products in a psoriasis plaque test and a murine psoriasis xenograft model. AB - The aim of the present study was to compare the effects of Daivobet and calcipotriol on clinical score and biomarker responses in a modified version of the Scholtz-Dumas psoriasis plaque assay. Furthermore, it was the aim to compare the effects of calcipotriol and betamethasone in the murine psoriasis xenograft model. Twenty four patients with psoriasis were treated topically once daily for three weeks, whereas the grafted mice were treated for four weeks. Clinical responses were scored twice weekly and biopsies were taken at the end of each study to analyse for skin biomarkers by histology and immunohistochemistry. The results clearly demonstrate effects on both clinical signs and biomarkers. In the patient study the total clinical score was reduced significantly with both Daivobet and calcipotriol. Both treatments reduced epidermal thickness, Ki-67 and cytokeratin 16 expression. T cell infiltration was significantly reduced by Daivobet but only marginally by calcipotriol. Both treatments showed strong effects on the epidermal psoriatic phenotype.Results from the xenograft model essentially showed the same results. However differences were observed when investigating subtypes of T cells.The study demonstrates the feasibility of obtaining robust biomarker data in the psoriasis plaque test that correlate well with those obtained in other clinical studies. Furthermore, the biomarker data from the plaque test correlate with biopsy data from the grafted mice. PMID- 20017944 TI - Whole genome transcript profiling from fingerstick blood samples: a comparison and feasibility study. AB - BACKGROUND: Whole genome gene expression profiling has revolutionized research in the past decade especially with the advent of microarrays. Recently, there have been significant improvements in whole blood RNA isolation techniques which, through stabilization of RNA at the time of sample collection, avoid bias and artifacts introduced during sample handling. Despite these improvements, current human whole blood RNA stabilization/isolation kits are limited by the requirement of a venous blood sample of at least 2.5 mL. While fingerstick blood collection has been used for many different assays, there has yet to be a kit developed to isolate high quality RNA for use in gene expression studies from such small human samples. The clinical and field testing advantages of obtaining reliable and reproducible gene expression data from a fingerstick are many; it is less invasive, time saving, more mobile, and eliminates the need of a trained phlebotomist. Furthermore, this method could also be employed in small animal studies, i.e. mice, where larger sample collections often require sacrificing the animal. In this study, we offer a rapid and simple method to extract sufficient amounts of high quality total RNA from approximately 70 microl of whole blood collected via a fingerstick using a modified protocol of the commercially available Qiagen PAXgene RNA Blood Kit. RESULTS: From two sets of fingerstick collections, about 70 uL whole blood collected via finger lancet and capillary tube, we recovered an average of 252.6 ng total RNA with an average RIN of 9.3. The post-amplification yields for 50 ng of total RNA averaged at 7.0 ug cDNA. The cDNA hybridized to Affymetrix HG-U133 Plus 2.0 GeneChips had an average % Present call of 52.5%. Both fingerstick collections were highly correlated with r(2) values ranging from 0.94 to 0.97. Similarly both fingerstick collections were highly correlated to the venous collection with r(2) values ranging from 0.88 to 0.96 for fingerstick collection 1 and 0.94 to 0.96 for fingerstick collection 2. CONCLUSIONS: Our comparisons of RNA quality and gene expression data of the fingerstick method with traditionally processed sample workflows demonstrate excellent RNA quality from the capillary collection as well as very high correlations of gene expression data. PMID- 20017945 TI - Identification of a novel germ-line mutation in the TP53 gene in a Mexican family with Li-Fraumeni syndrome. AB - BACKGROUND: Germ-line mutations of the TP53 gene are known to cause Li-Fraumeni syndrome, an autosomal, dominantly inherited, high-penetrance cancer predisposition syndrome characterized by the occurrence of a variety of cancers, mainly soft tissue sarcomas, adrenocortical carcinoma, leukemia, breast cancer, and brain tumors. METHODS: Mutation analysis was based on Denaturing high performance liquid chromatography (DHPLC) screening of exons 2-11 of the TP53 gene, sequencing, and cloning of DNA obtained from peripheral blood lymphocytes. RESULTS: We report herein on Li Fraumeni syndrome in a family whose members are carriers of a novel TP53 gene mutation at exon 4. The mutation comprises an insertion/duplication of seven nucleotides affecting codon 110 and generating a new nucleotide sequence and a premature stop codon at position 150. With this mutation, the p53 protein that should be translated lacks the majority of the DNA binding domain. CONCLUSION: To our knowledge, this specific alteration has not been reported previously, but we believe it is the cause of the Li-Fraumeni syndrome in this family. PMID- 20017946 TI - Burkholderia thailandensis harbors two identical rhl gene clusters responsible for the biosynthesis of rhamnolipids. AB - BACKGROUND: Rhamnolipids are surface active molecules composed of rhamnose and beta-hydroxydecanoic acid. These biosurfactants are produced mainly by Pseudomonas aeruginosa and have been thoroughly investigated since their early discovery. Recently, they have attracted renewed attention because of their involvement in various multicellular behaviors. Despite this high interest, only very few studies have focused on the production of rhamnolipids by Burkholderia species. RESULTS: Orthologs of rhlA, rhlB and rhlC, which are responsible for the biosynthesis of rhamnolipids in P. aeruginosa, have been found in the non infectious Burkholderia thailandensis, as well as in the genetically similar important pathogen B. pseudomallei. In contrast to P. aeruginosa, both Burkholderia species contain these three genes necessary for rhamnolipid production within a single gene cluster. Furthermore, two identical, paralogous copies of this gene cluster are found on the second chromosome of these bacteria. Both Burkholderia spp. produce rhamnolipids containing 3-hydroxy fatty acid moieties with longer side chains than those described for P. aeruginosa. Additionally, the rhamnolipids produced by B. thailandensis contain a much larger proportion of dirhamnolipids versus monorhamnolipids when compared to P. aeruginosa. The rhamnolipids produced by B. thailandensis reduce the surface tension of water to 42 mN/m while displaying a critical micelle concentration value of 225 mg/L. Separate mutations in both rhlA alleles, which are responsible for the synthesis of the rhamnolipid precursor 3-(3-hydroxyalkanoyloxy)alkanoic acid, prove that both copies of the rhl gene cluster are functional, but one contributes more to the total production than the other. Finally, a double DeltarhlA mutant that is completely devoid of rhamnolipid production is incapable of swarming motility, showing that both gene clusters contribute to this phenotype. CONCLUSIONS: Collectively, these results add another Burkholderia species to the list of bacteria able to produce rhamnolipids and this, by the means of two identical functional gene clusters. Our results also demonstrate the very impressive tensio-active properties these long-chain rhamnolipids possess in comparison to the well-studied short-chain ones from P. aeruginosa. PMID- 20017947 TI - Over-expression of miR172 causes loss of spikelet determinacy and floral organ abnormalities in rice (Oryza sativa). AB - BACKGROUND: Regulation of gene expression by microRNAs (miRNAs) plays a crucial role in many developmental and physiological processes in plants. miRNAs act to repress expression of their target genes via mRNA cleavage or translational repression. Dozens of miRNA families have been identified in rice, 21 of which are conserved between rice and Arabidopsis. miR172 is a conserved miRNA family which has been shown to regulate expression of APETALA2 (AP2)-like transcription factors in Arabidopsis and maize. The rice genome encodes five AP2-like genes predicted to be targets of miR172. To determine whether these rice AP2-like genes are regulated by miR172 and investigate the function of the target genes, we studied the effect of over-expressing two members of the miR172 family on rice plant development. RESULTS: Analysis of miR172 expression showed that it is most highly expressed in late vegetative stages and developing panicles. Analyses of expression of three miR172 targets showed that SUPERNUMERARY BRACT (SNB) and Os03g60430 have high expression in developing panicles. Expression of miR172 was not inversely correlated with expression of its targets although miR172-mediated cleavage of SNB was detected by 5' rapid amplification of cDNA ends (RACE). Over expression of miR172b in rice delayed the transition from spikelet meristem to floral meristem, and resulted in floral and seed developmental defects, including changes to the number and identity of floral organs, lower fertility and reduced seed weight. Plants over-expressing miR172b not only phenocopied the T-DNA insertion mutant of SNB but showed additional defects in floret development not seen in the snb mutant. However SNB expression was not reduced in the miR172b over-expression plants. CONCLUSIONS: The phenotypes resulting from over expression of miR172b suggests it represses SNB and at least one of the other miR172 targets, most likely Os03g60430, indicating roles for other AP2-like genes in rice floret development. miR172 and the AP2-like genes had overlapping expression patterns in rice and their expression did not show an obvious negative correlation. There was not a uniform decrease in the expression of the AP2-like miR172 target mRNAs in the miR172b over-expression plants. These observations are consistent with miR172 functioning via translational repression or with expression of the AP2-like genes being regulated by a negative feedback loop. PMID- 20017948 TI - Modeling of longitudinal polytomous outcome from complex survey data--application to investigate an association between mental distress and non-malignant respiratory diseases. AB - BACKGROUND: The data from longitudinal complex surveys based on multi-stage sampling designs contain cross-sectional dependencies among units due to clustered nature of the data and within-subject dependencies due to repeated measurements. Special statistical methods are required to analyze longitudinal complex survey data. METHODS: Statistics Canada's longitudinal National Population Health Survey (NPHS) dataset from the first five cycles (1994/1995 to 2002/2003) was used to investigate the effects of demographic, social, life style, and health-related factors on the longitudinal changes of mental distress scores among the NPHS participants who self-reported physician diagnosed respiratory diseases, specifically asthma and chronic bronchitis. The NPHS longitudinal sample includes 17,276 persons of all ages. In this report, participants 15 years and older (n = 14,713) were considered for statistical analysis. Mental distress, an ordinal outcome variable (categories: no/low, moderate, and high) was examined. Ordered logistic regression models based on the weighted generalized estimating equations approach were fitted to investigate the association between respiratory diseases and mental distress adjusting for other covariates of interest. Variance estimates of regression coefficients were computed by using bootstrap methods. The final model was used to predict the probabilities of prevalence of no/low, moderate or high mental distress scores. RESULTS: Accounting for design effects does not vary the significance of the coefficients of the model. Participants suffering with chronic bronchitis were significantly at a higher risk (ORadj = 1.37; 95% CI: 1.12-1.66) of reporting high levels of mental distress compared to those who did not self-report chronic bronchitis. There was no significant association between asthma and mental distress. There was a significant interaction between sex and self-perceived general health status indicating a dose-response relationship. Among females, the risk of mental distress increases with increasing deteriorating (from excellent to very poor) self-perceived general health. CONCLUSIONS: A positive association was observed between the physician diagnosed self-reported chronic bronchitis and an increased prevalence of mental distress when adjusted for important covariates. Variance estimates of regression coefficients obtained from the sandwich estimator (i.e. not accounting for design effects) were similar to bootstrap variance estimates (i.e. accounting for design effects). Even though these two sets of variance estimates are similar, it is more appropriate to use bootstrap variance estimates. PMID- 20017949 TI - Acute dosing of latrepirdine (Dimebon), a possible Alzheimer therapeutic, elevates extracellular amyloid-beta levels in vitro and in vivo. AB - BACKGROUND: Recent reports suggest that latrepirdine (Dimebon, dimebolin), a retired Russian antihistamine, improves cognitive function in aged rodents and in patients with mild to moderate Alzheimer's disease (AD). However, the mechanism(s) underlying this benefit remain elusive. AD is characterized by extracellular accumulation of the amyloid-beta (Abeta) peptide in the brain, and Abeta-lowering drugs are currently among the most popular anti-amyloid agents under development for the treatment of AD. In the current study, we assessed the effect of acute dosing of latrepirdine on levels of extracellular Abeta using in vitro and in vivo experimental systems. RESULTS: We evaluated extracellular levels of Abeta in three experimental systems, under basal conditions and after treatment with latrepirdine. Mouse N2a neuroblastoma cells overexpressing Swedish APP were incubated for 6 hr in the presence of either vehicle or vehicle + latrepirdine (500pM-5 muM). Synaptoneurosomes were isolated from TgCRND8 mutant APP-overexpressing transgenic mice and incubated for 0 to 10 min in the absence or presence of latrepirdine (1 muM or 10 muM). Drug-naive Tg2576 Swedish mutant APP overexpressing transgenic mice received a single intraperitoneal injection of either vehicle or vehicle + latrepirdine (3.5 mg/kg). Picomolar to nanomolar concentrations of acutely administered latrepirdine increased the extracellular concentration of Abeta in the conditioned media from Swedish mutant APP overexpressing N2a cells by up to 64% (p = 0.01), while a clinically relevant acute dose of latrepirdine administered i.p. led to an increase in the interstitial fluid of freely moving APP transgenic mice by up to 40% (p = 0.01). Reconstitution of membrane protein trafficking and processing is frequently inefficient, and, consistent with this interpretation, latrepirdine treatment of isolated TgCRND8 synaptoneurosomes involved higher concentrations of drug (1-10 muM) and led to more modest increases in extracellular Abeta(x-42 )levels (+10%; p = 0.001); of note, however, was the observation that extracellular Abeta(x-40 )levels did not change. CONCLUSIONS: Here, we report the surprising association of acute latrepirdine dosing with elevated levels of extracellular Abeta as measured in three independent neuron-related or neuron-derived systems, including the hippocampus of freely moving Tg2576 mice. Given the reported association of chronic latrepirdine treatment with improvement in cognitive function, the effects of chronic latrepirdine treatment on extracellular Abeta levels must now be determined. PMID- 20017950 TI - Emergency treatment of complicated incisional hernias: a case study. AB - BACKGROUND: The emergency treatment of incisional hernias is infrequent but it can be complicated with strangulation or obstruction and in some cases the surgical approach may also include an intestinal resection with the possibility of peritoneal contamination. Our study aims at reporting our experience in the emergency treatment of complicated incisional hernias. METHODS: Since January 1999 till July 2008, 89 patients (55 males and 34 females) were treated for complicated incisional hernias in emergency. The patients were divided in two groups: Group I consisting of 33 patients that were treated with prosthesis apposition and Group II, consisting of 56 patients that were treated by performing a direct abdominal wall muscles suture. RESULTS: All the patients underwent a 6-month follow up; we noticed 9 recurrences (9/56, 16%) in the patients treated with direct abdominal wall muscles suture and 1 recurrence (1/33, 3%) in the group of patients treated with the prosthesis apposition. CONCLUSIONS: According to our experience, the emergency treatment of complicated incisional hernias through prosthesis apposition is always feasible and ensures less post-operative complications (16% vs 21,2%) and recurrences (3% vs 16%) compared to the patients treated with direct muscular suture. PMID- 20017951 TI - Clonal dissemination of the multi-drug resistant Salmonella enterica serovar Braenderup, but not the serovar Bareilly, of prevalent serogroup C1 Salmonella from Taiwan. AB - BACKGROUND: Nontyphoidal Salmonella is the main cause of human salmonellosis. In order to study the prevalent serogroups and serovars of clinical isolates in Taiwan, 8931 Salmonellae isolates were collected from 19 medical centers and district hospitals throughout the country from 2004 to 2007. The pulsed-field eletrophoresis types (PFGE) and antibiotic resistance profiles of Salmonella enterica serovars Bareilly (S. Bareilly) and Braenderup (S. Braenderup) were compared, and multi-drug resistance (MDR) plasmids were characterized. RESULTS: Over 95% of human salmonellosis in Taiwan was caused by five Salmonella serogroups: B, C1, C2-C3, D1, and E1. S. Typhymurium, S. Enteritidis, S. Stanley and S. Newport were the four most prevalent serovars, accounting for about 64% of isolates. While only one or two major serovars from four of the most prevalent serogroups were represented, four predominant serovars were found in serogroup C1 Salmonellae. The prevalence was decreasing for S. Choleraeuis and S. Braenderup, and S. Virchow and increasing for S. Bareilly. S. Braenderup mainly caused gastroenteritis in children; in contrast, S. Bareiley infected children and elderly people. Both serovars differed by XbaI-PFGE patterns. Almost all S. Bareilly isolates were susceptible to antibiotics of interest, while all lacked plasmids and belonged to one clone. Two distinct major clones in S. Braenderup were cluster A, mainly including MDR isolates with large MDR plasmid from North Taiwan, and cluster B, mainly containing susceptible isolates without R plasmid from South Taiwan. In cluster A, there were two types of conjugative R plasmids with sizes ranging from 75 to 130 kb. Type 1 plasmids consisted of replicons F1A/F1B, blaTEM, IS26, and a class 1 integron with the genes dfrA12-orfF-aadA2 qacEDelta1-sulI. Type 2 plasmids belonged to incompatibility group IncI, contained tnpA-blaCMY-2-blc-sugE genetic structures and lacked both IS26 and class 1 integrons. Although type 2 plasmids showed higher conjugation capability, type 1 plasmids were the predominant plasmid. CONCLUSIONS: Serogroups B, C1, C2 C3, D1, and E1 of Salmonella caused over 95% of human salmonellosis. Two prevalent serovars within serogroup C1, S. Bareilly and cluster B of S. Braenderup, were clonal and drug-susceptible. However, cluster A of S. Braenderup was MDR and probably derived from susceptible isolates by acquiring one of two distinct conjugative R plasmids. PMID- 20017952 TI - Distinct functions of HTLV-1 Tax1 from HTLV-2 Tax2 contribute key roles to viral pathogenesis. AB - While the human T-cell leukemia virus type 1 (HTLV-1) is the etiologic agent of adult T-cell leukemia/lymphoma (ATL), to date, its close relative HTLV-2 is not associated with ATL or other types of malignancies. Accumulating evidence shows that HTLV-1 Tax1 and HTLV-2 Tax2 have many shared activities, but the two proteins have a limited number of significantly distinct activities, and these distinctions appear to play key roles in HTLV-1 specific pathogenesis. In this review, we summarize the functions of Tax1 associated with cell survival, cell proliferation, persistent infection as well as pathogenesis. We emphasize special attention to distinctions between Tax1 and Tax2. PMID- 20017953 TI - Characterization of the stem cell system of the acoel Isodiametra pulchra. AB - BACKGROUND: Tissue plasticity and a substantial regeneration capacity based on stem cells are the hallmark of several invertebrate groups such as sponges, cnidarians and Platyhelminthes. Traditionally, Acoela were seen as an early branching clade within the Platyhelminthes, but became recently positioned at the base of the Bilateria. However, little is known on how the stem cell system in this new phylum is organized. In this study, we wanted to examine if Acoela possess a neoblast-like stem cell system that is responsible for development, growth, homeostasis and regeneration. RESULTS: We established enduring laboratory cultures of the acoel Isodiametra pulchra (Acoela, Acoelomorpha) and implemented in situ hybridization and RNA interference (RNAi) for this species. We used BrdU labelling, morphology, ultrastructure and molecular tools to illuminate the morphology, distribution and plasticity of acoel stem cells under different developmental conditions. We demonstrate that neoblasts are the only proliferating cells which are solely mesodermally located within the organism. By means of in situ hybridisation and protein localisation we could demonstrate that the piwi-like gene ipiwi1 is expressed in testes, ovaries as well as in a subpopulation of somatic stem cells. In addition, we show that germ cell progenitors are present in freshly hatched worms, suggesting an embryonic formation of the germline. We identified a potent stem cell system that is responsible for development, homeostasis, regeneration and regrowth upon starvation. CONCLUSIONS: We introduce the acoel Isodiametra pulchra as potential new model organism, suitable to address developmental questions in this understudied phylum. We show that neoblasts in I. pulchra are crucial for tissue homeostasis, development and regeneration. Notably, epidermal cells were found to be renewed exclusively from parenchymally located stem cells, a situation known only from rhabditophoran flatworms so far. For further comparison, it will be important to analyse the stem cell systems of other key-positioned understudied taxa. PMID- 20017954 TI - The transcription factor RBP-J is essential for retinal cell differentiation and lamination. AB - BACKGROUND: The highly ordered vertebrate retina is composed of seven cell types derived from a common pool of retinal progenitor cells (RPCs), and is a good model for the studies of cell differentiation and interaction during neural development. Notch signaling plays a pivotal role in retinogenesis in mammals, but the full scope of the functions of Notch pathway, and the underlying molecular mechanisms, remain unclear. RESULTS: In this study, we conditionally knocked out RBP-J, the critical transcription factor downstream to all four Notch receptors, in RPCs of mouse retina at different developmental stages. Disruption of RBP-J at early retinogenesis resulted in accelerated RPCs differentiation, but only photoreceptors and ganglion cells were overrepresented, with other neuronal populations diminished. Similarly, deletion of RBP-J at early postnatal days also led to overproduction of photoreceptors, suggesting that RBP-J governed RPCs specification and differentiation through retinogenesis. In all the RBP-J deletion models, the retinal laminar structures were distorted by the formation of numerous rosette-like structures, reminiscent of beta-catenin deficient retina. Indeed, we found that these rosettes aligned with gaps in beta-catenin expression at the apical surface of the retina. By in vivo electroporation mediated transfection, we demonstrated that lamination defects in RBP-J deficient retinae were rescued by overexpressing beta-catenin. CONCLUSIONS: Our data indicate that RBP-J-mediated canonical Notch signaling governs retinal cell specification and differentiation, and maintains retinal lamination through the expression of beta-catenin. PMID- 20017955 TI - Inhibiting toll-like receptor 4 signaling ameliorates pulmonary fibrosis during acute lung injury induced by lipopolysaccharide: an experimental study. AB - BACKGROUND: Toll-like receptor 4 (TLR4) is essential in lipopolysaccharide (LPS) induced fibroblast activation and collagen secretion in vitro. However, its effects on the process of lung fibroblast activation and fibrosis initiation during LPS induced acute lung injury (ALI) remain unknown. The goal of the present study was to determine the effect of inhibiting TLR4 on LPS-induced ALI and fibrosis in vivo. METHODS: The ALI model was established by intraperitoneal injection of LPS in mice. TLR4-small hairpin RNA (shRNA) lentivirus was injected intravenously into the mice to inhibit TLR4 expression. mRNA and protein levels were detected by real-time PCR and Western-blot analysis, respectively. The contents of the C-terminal propeptide of type I procollagen (PICP) in bronchoalveolar lavage fluid (BALF) were detected by ELISA, and the degree of fibrosis was detected by van Gieson collagen staining, the hydroxyproline assay, and alpha smooth muscle actin (alpha-SMA) immunohistochemical staining. RESULTS: Overexpression of TLR4, type I procollagen, alpha-SMA, and p-AKT in murine pulmonary tissue after intraperitoneal injection of LPS at 72 hours and 28 days were detected. Moreover, the degree of fibrosis was shown to increase by ELISA analysis of PICP in BALF, van Gieson collagen staining, the hydroxyproline assay, and alpha-SMA immunohistochemical staining. All of these changes were alleviated by intravenous infection with TLR4-shRNA lentivirus. CONCLUSIONS: Inhibiting TLR4 signaling could ameliorate fibrosis at the early stage of ALI induced by LPS. PMID- 20017957 TI - A practical comparison of methods for detecting transcription factor binding sites in ChIP-seq experiments. AB - BACKGROUND: Chromatin immunoprecipitation coupled with massively parallel sequencing (ChIP-seq) is increasingly being applied to study transcriptional regulation on a genome-wide scale. While numerous algorithms have recently been proposed for analysing the large ChIP-seq datasets, their relative merits and potential limitations remain unclear in practical applications. RESULTS: The present study compares the state-of-the-art algorithms for detecting transcription factor binding sites in four diverse ChIP-seq datasets under a variety of practical research settings. First, we demonstrate how the biological conclusions may change dramatically when the different algorithms are applied. The reproducibility across biological replicates is then investigated as an internal validation of the detections. Finally, the predicted binding sites with each method are compared to high-scoring binding motifs as well as binding regions confirmed in independent qPCR experiments. CONCLUSIONS: In general, our results indicate that the optimal choice of the computational approach depends heavily on the dataset under analysis. In addition to revealing valuable information to the users of this technology about the characteristics of the binding site detection approaches, the systematic evaluation framework provides also a useful reference to the developers of improved algorithms for ChIP-seq data. PMID- 20017958 TI - A structured framework for improving outbreak investigation audits. AB - BACKGROUND: Outbreak investigation is a core function of public health agencies. Suboptimal outbreak investigation endangers both public health and agency reputations. While audits of clinical medical and nursing practice are conducted as part of continuous quality improvement, public health agencies rarely make systematic use of structured audits to ensure best practice for outbreak responses, and there is limited guidance or policy to guide outbreak audit. METHODS: A framework for prioritising which outbreak investigations to audit, an approach for conducting a successful audit, and a template for audit trigger questions was developed and trialled in four foodborne outbreaks and a respiratory disease outbreak in Australia. RESULTS: The following issues were identified across several structured audits: the need for clear definitions of roles and responsibilities both within and between agencies, improved communication between agencies and with external stakeholders involved in outbreaks, and the need for development of performance standards in outbreak investigations - particularly in relation to timeliness of response. Participants considered the audit process and methodology to be clear, useful, and non threatening. Most audits can be conducted within two to three hours, however, some participants felt this limited the scope of the audit. CONCLUSION: The framework was acceptable to participants, provided an opportunity for clarifying perceptions and enhancing partnership approaches, and provided useful recommendations for approaching future outbreaks. Future challenges include incorporating feedback from broader stakeholder groups, for example those of affected cases, institutions and businesses; assessing the quality of a specific audit; developing training for both participants and facilitators; and building a central capacity to support jurisdictions embarking on an audit. The incorporation of measurable performance criteria or sharing of benchmark performance criteria will assist in the standardisation of outbreak investigation audit and further quality improvement. PMID- 20017956 TI - Compound K, a metabolite of ginseng saponin, induces apoptosis via caspase-8 dependent pathway in HL-60 human leukemia cells. AB - BACKGROUND: Compound K [20-O-beta-(D-glucopyranosyl)-20(S)-protopanaxadiol], a metabolite of the protopanaxadiol-type saponins of Panax ginseng C.A. Meyer, has been reported to possess anti-tumor properties to inhibit angiogenesis and to induce tumor apoptosis. In the present study, we investigated the effect of Compound K on apoptosis and explored the underlying mechanisms involved in HL-60 human leukemia cells. METHODS: We examined the effect of Compound K on the viabilities of various cancer cell lines using MTT assays. DAPI assay, Annexin V and PI double staining, Western blot assay and immunoprecipitation were used to determine the effect of Compound K on the induction of apoptosis. RESULTS: Compound K was found to inhibit the viability of HL-60 cells in a dose- and time dependent manner with an IC50 of 14 muM. Moreover, this cell death had typical features of apoptosis, that is, DNA fragmentation, DNA ladder formation, and the externalization of Annexin V targeted phosphatidylserine residues in HL-60 cells. In addition, compound-K induced a series of intracellular events associated with both the mitochondrial- and death receptor-dependent apoptotic pathways, namely, (1) the activation of caspases-3, -8, and -9; (2) the loss of mitochondrial membrane potential; (3) the release of cytochrome c and Smac/DIABLO to the cytosol; (4) the translocation of Bid and Bax to mitochondria; and (5) the downregulations of Bcl-2 and Bcl-xL. Furthermore, a caspase-8 inhibitor completely abolished caspase-3 activation, Bid cleavage, and subsequent DNA fragmentation by Compound K. Interestingly, the activation of caspase-3 and -8 and DNA fragmentation were significantly prevented in the presence of cycloheximide, suggesting that Compound K-induced apoptosis is dependent on de novo protein synthesis. CONCLUSIONS: The results indicate that caspase-8 plays a key role in Compound K-stimulated apoptosis via the activation of caspase-3 directly or indirectly through Bid cleavage, cytochrome c release, and caspase-9 activation. PMID- 20017960 TI - Recursive partitioning analysis of prognostic factors in WHO grade III glioma patients treated with radiotherapy or radiotherapy plus chemotherapy. AB - BACKGROUND: We evaluated the hierarchical risk groups for the estimated survival of WHO grade III glioma patients using recursive partitioning analysis (RPA). To our knowledge, this is the first study to address the results of RPA specifically for WHO grade III gliomas. METHODS: A total of 133 patients with anaplastic astrocytoma (AA, n = 56), anaplastic oligodendroglioma (AO, n = 67), or anaplastic oligoastrocytoma (AOA, n = 10) were included in the study. These patients were treated with either radiotherapy alone or radiotherapy followed by PCV chemotherapy after surgery. Five prognostic factors, including histological subsets, age, performance status, extent of resection, and treatment modality were incorporated into the RPA. The final nodes of RPA were grouped according to their survival times, and the Kaplan-Meier graphs are presented as the final set of prognostic groups. RESULTS: Four risk groups were defined based on the clinical prognostic factors excluding age, and split variables were all incorporated into the RPA. Survival analysis showed significant differences in mean survival between the different groups: 163.4 months (95% CI: 144.9-182.0), 109.5 months (86.7-132.4), 66.6 months (50.8-82.4), and 27.7 months (16.3-39.0), respectively, from the lowest to the highest risk group (p = 0.00). CONCLUSION: The present study shows that RPA grouping with clinical prognostic factors can successfully predict the survival of patients with WHO grade III glioma. PMID- 20017959 TI - Depot-specific differences in perilipin and hormone-sensitive lipase expression in lean and obese. AB - BACKGROUND: Mainly dependent on hormone-sensitive lipase, lipolysis is differently impaired between fat depots in human obesity. Perilipin A expression is a critical element in adipocyte lipolysis. The present study aimed at comparing expression and subcellular distribution of perilipin and hormone sensitive lipase in two abdominal adipose tissues of lean and obese women. We examined whether regional differences in perilipin expression contribute to impaired lipolytic rates. METHODS: Abdominal subcutaneous and omental adipose tissues were obtained from six lean and ten obese women. We measured total protein content and relative distribution of hormone-sensitive lipase and perilipin proteins between lipid and non-lipid fractions in tissue homogenates. Hormone-sensitive lipase and perilipin mRNA levels, adipocyte size, basal (non stimulated) and noradrenaline-stimulated lipolysis in isolated adipocytes were determined. RESULTS: Adipocytes were significantly larger in the obese versus the lean women and in subcutaneous versus omental fat. Expressed as a function of cell number, basal lipolysis and noradrenaline responsiveness were higher in subcutaneous versus omental adipocytes from the obese women (P < 0.05). Despite higher or identical mRNA levels in the lean and the obese subjects and in subcutaneous and omental tissues, perilipin protein expression was lower in both depots in the obese versus the lean women, and in subcutaneous versus omental in both lean and obese women (P < 0.05). Perilipin was mostly (above 80%) present in the lipid fraction in both depots from the obese patients and the value decreased to 60% in the lean subjects (P < 0.05). Perilipin protein expression was inversely correlated to adipocyte size and basal lipolysis in both depots. Despite higher mRNA levels, hormone-sensitive lipase protein expression decreased in both depots of the obese women. Regional difference for hormone-sensitive lipase was reported in lipid fraction of subcutaneous fat of the obese subjects: hormone-sensitive lipase content was twice as low as in omental adipose tissue. CONCLUSION: In both fat depots, a reduced perilipin protein expression was observed in women obesity. Perilipin protein level may contribute to differences in basal lipolysis and in adipocyte size between fat depots and may regulate lipid accumulation in adipocytes. Differences in hormone-sensitive lipase subcellular distribution were reported between fat depots in the obese women. PMID- 20017962 TI - Genetic Analysis Workshop 16: Strategies for genome-wide association study analyses. PMID- 20017961 TI - Oxidative stress promotes autophagic cell death in human neuroblastoma cells with ectopic transfer of mitochondrial PPP2R2B (Bbeta2). AB - BACKGROUND: The multifunctional protein phosphatase 2A (PP2A) is a heterotrimeric serine/threonine protein phosphatase composed of a scaffolding, catalytic and regulatory subunits. By modifying various downstream signal transducers, the aberrant expression of the brain-targeted regulatory subunit PPP2R2B is associated with the onset of a panel of neuronal disorders. The alternatively splicing of PPP2R2B encodes two regulatory subunit isoforms that determine cellular distribution of the neuron-specific holoenzyme to mitochondria (Bbeta2) and cytoplasm (Bbeta1), respectively. RESULTS: Human neuroblastoma cells were transfected with PPP2R2B constructs encoding the complete sequences of Bbeta2 and Bbeta1, respectively. The colonies with antibiotic resistance were selected as stable cell lines. Both ectopic Bbeta1 and Bbeta2 clones exhibited characteristics of autophagy. To test how cells respond to reactive oxygen species generators, the cells were treated with either hydrogen peroxide or t butyl hydroperoxide and Bbeta2 clones induced cell death. Suppression of autophagy using either RNA interference of the essential autophagy gene or pharmacological inhibitor rescued cell death caused by oxidative stress. CONCLUSIONS: Cells with ectopically expressed mitochondria-targeted regulatory subunit PPP2R2B of the holoenzyme PP2A were shown predisposed to autophagy and oxidative stress induced cell death that is related to apoptosis. The results promised a model for studying the mechanism and function of aberrant PPP2R2B expression in neuronal cells. The work provided a new target for understanding and prevention of neuropathogenesis. PMID- 20017963 TI - Representation of genetic association via attributable familial relative risks in order to identify polymorphisms functionally relevant to rheumatoid arthritis. AB - The results from association studies are usually summarized by a measure of evidence of association (frequentist or Bayesian probability values) that does not directly reflect the impact of the detected signals on familial aggregation. This article investigates the possible advantage of a two-dimensional representation of genetic association in order to identify polymorphisms relevant to disease: a measure of evidence of association (the Bayes factor, BF) combined with the estimated contribution to familiality (the attributable sibling relative risk, lambdas). Simulation and data from the North American Rheumatoid Consortium (NARAC) were used to assess the possible benefit under several scenarios. Simulation indicated that the allele frequencies to reach the maximum BF and the maximum attributable lambdas diverged as the size of the genetic effect increased. The representation of BF versus attributable lambdas for selected regions of NARAC data revealed that SNPs involved in replicated associations clearly departed from the bulk of SNPs in these regions. In the 12 investigated regions, and particularly in the low-recombination major histocompatibility region, the ranking of SNPs according to BF differed from the ranking of SNPs according to attributable lambdas. The present results should be generalized using more extensive simulations and additional real data, but they suggest that a characterization of genetic association by both BF and attributable lambdas may result in an improved ranking of variants for further biological analyses. PMID- 20017964 TI - Genome-wide association and linkage analysis of quantitative traits: comparison of likelihood-ratio test and conditional score statistic. AB - Over the past decade, genetic analysis has shifted from linkage studies, which identify broad regions containing putative trait loci, to genome-wide association studies, which detect the association of a marker with a specific phenotype. Because linkage and association analysis provide complementary information, developing a method to combine these analyses may increase the power to detect a true association. In this paper we compare a linkage score and association score test as well as a newly proposed combination of these two scores with traditional linkage and association methods. PMID- 20017965 TI - Identifying association under a previous linkage peak on chromosome 16 for body mass index using cross-sectional and longitudinal data of the Framingham Heart Study. AB - We performed association analysis under a previous linkage peak on chromosome 16 with genome-wide single-nucleotide polymorphism (SNP) data to identify genetic variants underlying body mass index (BMI). Data from all subjects with baseline measures and a subgroup who had complete data at four selected time points from the Framingham Heart Study were analyzed. The cross-sectional measures include BMI at baseline for all subjects, as well as BMI at selected time points for the subgroup. The longitudinal measure is the within-subject mean of BMI for the subgroup at the four time points.Association analysis was first performed using PLINK after dividing large pedigrees into nuclear families. We then followed up the identified regions by variance-components methods as implemented in SOLAR using the extended pedigrees.The strongest evidence for associations were observed at 52.3 Mbp (PLINK p = 0.00002, QTLD p = 0.005), on the FTO gene, and at 48.1 Mbp (PLINK p = 0.002, QTLD p = 0.0006) on chromosome 16, which are directly under the previous identified linkage peak. This association was consistently observed for all samples at baseline, and for the subgroup at time point 2, 3, 4 and MEAN, both by PLINK and SOLAR. In addition, another SNP/region at 46.7 Mbp on same chromosome was found to be associated with several BMI measures in the subgroup. Fine-mapping with more markers provided further evidence for SNP association with BMI in the same region (at 52.4 Mbp, QTLD p = 0.0003).These results suggest the existence of genes/DNA variations in these regions that contribute to BMI variation. PMID- 20017966 TI - Contrasting identity-by-descent estimators, association studies, and linkage analyses using the Framingham Heart Study data. AB - We explored the utility of population- and pedigree-based analyses using the Framingham Heart Study genome-wide 50 k single-nucleotide polymorphism marker data provided for Genetic Analysis Workshop 16. Our aims were: 1) to compare identity-by-descent sharing estimates from variable amounts of data; 2) to apply each of these estimates to a case-control association study designed to control for relatedness among samples; and 3) to contrast these results to those obtained using model-based and model-free linkage analysis methods. PMID- 20017967 TI - Genome-wide association analyses of North American Rheumatoid Arthritis Consortium and Framingham Heart Study data utilizing genome-wide linkage results. AB - The power of genome-wide association studies can be improved by incorporating information from previous study findings, for example, results of genome-wide linkage analyses. Weighted false-discovery rate (FDR) control can incorporate genome-wide linkage scan results into the analysis of genome-wide association data by assigning single-nucleotide polymorphism (SNP) specific weights. Stratified FDR control can also be applied by stratifying the SNPs into high and low linkage strata. We applied these two FDR control methods to the data of North American Rheumatoid Arthritis Consortium (NARAC) study and the Framingham Heart Study (FHS), combining both association and linkage analysis results. For the NARAC study, we used linkage results from a previous genome scan of rheumatoid arthritis (RA) phenotype. For the FHS study, we obtained genome-wide linkage scores from the same 550 k SNP data used for the association analyses of three lipids phenotypes (HDL, LDL, TG). We confirmed some genes previously reported for association with RA and lipid phenotypes. Stratified and weighted FDR methods appear to give improved ranks to some of the replicated SNPs for the RA data, suggesting linkage scan results could provide useful information to improve genome-wide association studies. PMID- 20017968 TI - A method to correct for population structure using a segregation model. AB - To overcome the "spurious" association caused by population stratification in population-based association studies, we propose a principal-component based method that can use both family and unrelated samples at the same time. More specifically, we adapt the multivariate logistic model, which is often used in segregation analysis and can allow for the family correlation structure, for association analysis. To correct the effect of hidden population structure, the first ten principal-components calculated from the matrix of marker genotype data are incorporated as covariates in the model. To test for the association, the marker of interest is also incorporated as a covariate in the model. We applied the proposed method to the second generation (i.e., the Offspring Cohort), in the Genetic Analysis Workshop 16 Framingham Heart Study 50 k data set to evaluate the performance of the method. Although there may have been difficulty in the convergence while maximizing the likelihood function as indicated by a flat likelihood, the distribution of the empirical p-values for the test statistic does show that the method has a correct type I error rate whenever the variance covariance matrix of the estimates can be computed. PMID- 20017969 TI - Linkage disequilibrium of single-nucleotide polymorphism data: how sampling methods affect estimates of linkage disequilibrium. AB - Linkage disequilibrium (LD) is an important measure used in the analysis of single-nucleotide polymorphism (SNP) data. We used the Genetic Analysis Workshop 16 (GAW16) Framingham Heart Study 500 k SNP data to explore the effect of sampling methods on estimating of LD for SNP data. METHOD AND DATA: We found 332 trios in the GAW16 Framingham SNP data. Repeated random samples without replacement, of different sizes of trios and independent individuals, are drawn from these 332 trios. For each sample, the LD is calculated using the Haploview program for the chromosome 1 SNP data. Percents of D' > 0.8 and r2 > 0.8 are calculated for different distance bins based on the Haploview output. The results are summarized by sample size and sampling methods to give us an overall view of the effect of sample size and sampling methods on the LD estimation. RESULTS: Trios design gave stable estimates. A sample of 30 to 40 trios gave estimates of percent of LD > 0.8 very close to those from 332 trios. When independent individuals are used, the estimates are less stable and are different from those obtained from the 332 trios for both D' and r2, with larger differences for D'. CONCLUSION: Our results suggest that trio design gives a stable estimate of LD. Therefore it may be more suitable for LD analysis than using independent individuals. We must be cautious when comparing the LD estimates from trios, and those from independent individuals. PMID- 20017970 TI - Detecting population stratification using related individuals. AB - Although identification of cryptic population stratification is necessary for case/control association analyses, it is also vital for linkage analyses and family-based association tests when founder genotypes are missing. However, including related individuals in an analysis such as EIGENSTRAT can result in bias; using only founders or one individual per pedigree results in loss of data and inaccurate estimates of stratification. We examine a generalization of principal-component analyses to allow for the inclusion of related individuals by down-weighting the significance of individual comparisons. PMID- 20017971 TI - Assessing the impact of global versus local ancestry in association studies. AB - BACKGROUND: To account for population stratification in association studies, principal-components analysis is often performed on single-nucleotide polymorphisms (SNPs) across the genome. Here, we use Framingham Heart Study (FHS) Genetic Analysis Workshop 16 data to compare the performance of local ancestry adjustment for population stratification based on principal components (PCs) estimated from SNPs in a local chromosomal region with global ancestry adjustment based on PCs estimated from genome-wide SNPs. METHODS: Standardized height residuals from unrelated adults from the FHS Offspring Cohort were averaged from longitudinal data. PCs of SNP genotype data were calculated to represent individual's ancestry either 1) globally using all SNPs across the genome or 2) locally using SNPs in adjacent 20-Mbp regions within each chromosome. We assessed the extent to which there were differences in association studies of height depending on whether PCs for global, local, or both global and local ancestry were included as covariates. RESULTS: The correlations between local and global PCs were low (r < 0.12), suggesting variability between local and global ancestry estimates. Genome-wide association tests without any ancestry adjustment demonstrated an inflated type I error rate that decreased with adjustment for local ancestry, global ancestry, or both. A known spurious association was replicated for SNPs within the lactase gene, and this false-positive association was abolished by adjustment with local or global ancestry PCs. CONCLUSION: Population stratification is a potential source of bias in this seemingly homogenous FHS population. However, local and global PCs derived from SNPs appear to provide adequate information about ancestry. PMID- 20017972 TI - Principal-component-based population structure adjustment in the North American Rheumatoid Arthritis Consortium data: impact of single-nucleotide polymorphism set and analysis method. AB - Population structure occurs when a sample is composed of individuals with different ancestries and can result in excess type I error in genome-wide association studies. Genome-wide principal-component analysis (PCA) has become a popular method for identifying and adjusting for subtle population structure in association studies. Using the Genetic Analysis Workshop 16 (GAW16) NARAC data, we explore two unresolved issues concerning the use of genome-wide PCA to account for population structure in genetic associations studies: the choice of single nucleotide polymorphism (SNP) subset and the choice of adjustment model. We computed PCs for subsets of genome-wide SNPs with varying levels of LD. The first two PCs were similar for all subsets and the first three PCs were associated with case status for all subsets. When the PCs associated with case status were included as covariates in an association model, the reduction in genomic inflation factor was similar for all SNP sets. Several models have been proposed to account for structure using PCs, but it is not yet clear whether the different methods will result in substantively different results for association studies with individuals of European descent. We compared genome-wide association p values and results for two positive-control SNPs previously associated with rheumatoid arthritis using four PC adjustment methods as well as no adjustment and genomic control. We found that in this sample, adjusting for the continuous PCs or adjusting for discrete clusters identified using the PCs adequately accounts for the case-control population structure, but that a recently proposed randomization test performs poorly. PMID- 20017973 TI - Comparison of methods for correcting population stratification in a genome-wide association study of rheumatoid arthritis: principal-component analysis versus multidimensional scaling. AB - Population stratification (PS) represents a major challenge in genome-wide association studies. Using the Genetic Analysis Workshop 16 Problem 1 data, which include samples of rheumatoid arthritis patients and healthy controls, we compared two methods that can be used to evaluate population structure and correct PS in genome-wide association studies: the principal-component analysis method and the multidimensional-scaling method. While both methods identified similar population structures in this dataset, principal-component analysis performed slightly better than the multidimensional-scaling method in correcting for PS in genome-wide association analysis of this dataset. PMID- 20017974 TI - Simultaneous analysis of all single-nucleotide polymorphisms in genome-wide association study of rheumatoid arthritis. AB - The availability of very large number of markers by modern technology makes genome-wide association studies very popular. The usual approach is to test single-nucleotide polymorphisms (SNPs) one at a time for association with disease status. However, it may not be possible to detect marginally significant effects by single-SNP analysis. Simultaneous analysis of SNPs enables detection of even those SNPs with small effect by evaluating the collective impact of several neighboring SNPs. Also, false-positive signals may be weakened by the presence of other neighboring SNPs included in the analysis. We analyzed the North American Rheumatoid Arthritis Consortium data of Genetic Analysis Workshop 16 using HLasso, a new method for simultaneous analysis of SNPs. The simultaneous analysis approach has excellent control of type I error, and many of the previously reported results of single-SNP analyses were confirmed by this approach. PMID- 20017975 TI - Graphic analysis of population structure on genome-wide rheumatoid arthritis data. AB - Principal-component analysis (PCA) has been used for decades to summarize the human genetic variation across geographic regions and to infer population migration history. Reduction of spurious associations due to population structure is crucial for the success of disease association studies. Recently, PCA has also become a popular method for detecting population structure and correction of population stratification in disease association studies. Inspired by manifold learning, we propose a novel method based on spectral graph theory. Regarding each study subject as a node with suitably defined weights for its edges to close neighbors, one can form a weighted graph. We suggest using the spectrum of the associated graph Laplacian operator, namely, Laplacian eigenfunctions, to infer population structures instead of principal components (PCs). For the whole genome wide association data for the North American Rheumatoid Arthritis Consortium (NARAC) provided by Genetic Workshop Analysis 16, Laplacian eigenfunctions revealed more meaningful structures of the underlying population than PCA. The proposed method has connection to PCA, and it naturally includes PCA as a special case. Our simple method is computationally fast and is suitable for disease studies at the genome-wide scale. PMID- 20017976 TI - Accommodating population stratification in case-control association analysis: a new test and its application to genome-wide study on rheumatoid arthritis. AB - It is well known that conventional association tests can lead to excessive false positives when there is population stratification. We propose a new test for detecting genetic association with a case-control study design. Unlike some other methods for handling population stratification, we treat the cases as a population and the controls as another one even though each of them may be a mixture of several sub-populations. A likelihood-ratio test is used to test whether the allele frequency of a testing single-nucleotide polymorphism in the case population is the same as that in the control population. This new test is applied to the Genetic Analysis Workshop 16 Problem 1 data on rheumatoid arthritis. Compared with the Pearson chi-square genotype test, the association strength of many single-nucleotide polymorphisms is decreased while the signal at the HLA region on 6p21 is maintained. PMID- 20017978 TI - Influence of control selection in genome-wide association studies: the example of diabetes in the Framingham Heart Study. AB - Epidemiologic study designs represent a major challenge for genome-wide association studies. Most such studies to date have selected controls from the pool of participants without the disease of interest at the end of the study time. These choices can lead to biased estimates of exposure effects. Using data from the Framingham Heart Study (Genetic Analysis Workshop 16 Problem 2), we evaluate the impact on genetic association estimates for designs with control selection based on status at the end of a study (case exclusion (CE) sampling) to control selection based on incidence density (ID) sampling, when controls are selected from the pool of participants who are disease-free at the time a case is diagnosed. Cases are defined as those diagnosed with type 2 diabetes (T2D). We estimated odds ratios for 18 previously confirmed T2D variants using 189 cases selected by ID sampling and using 231 cases selected by CE sampling. We found none of these single-nucleotide polymorphisms to be significantly associated with T2D using either design. Because these empirical analyses were based on a small number of cases and on single-nucleotide polymorphisms with likely small effect sizes, we supplemented this work with simulated data sets of 500 cases from each strategies across a variety of allele frequencies and effect sizes. In our simulated datasets, we show ID sampling to be less biased than CE, although CE shows apparent increased power due to the upward bias of point estimates. We conclude that ID sampling is an appropriate option for genome-wide association studies. PMID- 20017977 TI - Growth mixture modeling as an exploratory analysis tool in longitudinal quantitative trait loci analysis. AB - We examined the properties of growth mixture modeling in finding longitudinal quantitative trait loci in a genome-wide association study. Two software packages are commonly used in these analyses: Mplus and the SAS TRAJ procedure. We analyzed the 200 replicates of the simulated data with these programs using three tests: the likelihood-ratio test statistic, a direct test of genetic model coefficients, and the chi-square test classifying subjects based on the trajectory model's posterior Bayesian probability. The Mplus program was not effective in this application due to its computational demands. The distributions of these tests applied to genes not related to the trait were sensitive to departures from Hardy-Weinberg equilibrium. The likelihood-ratio test statistic was not usable in this application because its distribution was far from the expected asymptotic distributions when applied to markers with no genetic relation to the quantitative trait. The other two tests were satisfactory. Power was still substantial when we used markers near the gene rather than the gene itself. That is, growth mixture modeling may be useful in genome-wide association studies. For markers near the actual gene, there was somewhat greater power for the direct test of the coefficients and lesser power for the posterior Bayesian probability chi-square test. PMID- 20017979 TI - Growth mixture modelling in families of the Framingham Heart Study. AB - Growth mixture modelling, a less explored method in genetic research, addresses unobserved heterogeneity in population samples. We applied this technique to longitudinal data of the Framingham Heart Study. We examined systolic blood pressure (BP) measures in 1060 males from 692 families and detected three subclasses, which varied significantly in their developmental trajectories over time. The first class consisted of 60 high-risk individuals with elevated BP early in life and a steep increase over time. The second group of 131 individuals displayed first normal BP, but showed a significant increase over time and reached high BP values late in their life time. The largest group of 869 individuals could be considered a normative group with normal BP on all exams. To identify genetic modulators for this phenotype, we tested 2,340 single-nucleotide polymorphisms on chromosome 8 for association with the class membership probabilities of our model. The probability of being in Class 1 was significantly associated with a very rare variant (rs1445404) present in only four individuals from four different families located in the coding region of the gene EYA (eyes absent homolog 1 in Drosophila) (p = 1.39 x 10-13). Mutations in EYA are known to cause brachio-oto-renal syndrome, as well as isolated renal malformations. Renal malformations could cause high BP early in life. This result awaits replication; however, it suggests that analyzing genetic data stratified for high-risk subgroups defined by a unique development over time could be useful for the detection of rare mutations in common multi-factorial diseases. PMID- 20017980 TI - A multilevel linear mixed model of the association between candidate genes and weight and body mass index using the Framingham longitudinal family data. AB - Obesity has become an epidemic in many countries and is one of the major risk conditions for disease including type 2 diabetes, coronary heart disease, stroke, dyslipidemia, and hypertension. Recent genome-wide association studies have identified two genes (FTO and near MC4R) that were unequivocally associated with body mass index (BMI) and obesity. For the Genetic Analysis Workshop 16, data from the Framingham Heart Study were made available, including longitudinal anthropometric and metabolic traits for 7130 Caucasian individuals over three generations, each with follow-up data at up to four time points. We explored the associations between four single-nucleotide polymorphisms (SNPs) on FTO (rs1121980, rs9939609) or near MC4R (rs17782313, rs17700633) with weight and BMI under an additive model. We applied multilevel linear mixed model for continuous outcomes, using the Affymetrix 500k genome-wide genotype data for the four SNPs. The results of the multilevel modeling in the entire sample indicated that the minor alleles of the four SNPs were associated with higher weight and higher BMI. The most significant associations were between rs9939609 and weight (p = 4.7 x 10 6) and BMI (p = 8.9 x 10-8). The results also showed that, for SNPs at FTO, the homozygotes for the minor allele had the most pronounced increase in weight and BMI, while the common allele homozygotes gained less weight and BMI during the follow-up period. Linkage disequilibrium (LD) between the two FTO SNPs was strong (D' = 0.997, r2 = 0.875) but their haplotype was not significantly associated with either weight or BMI. The two SNPs near MC4R were in weak LD (D' = 0.487, r2 = 0.166). PMID- 20017981 TI - Longitudinal trends in the association of metabolic syndrome with 550 k single nucleotide polymorphisms in the Framingham Heart Study. AB - We investigated the association of metabolic syndrome (MetS) with a 500 k and a 50 k single-nucleotide polymorphism (SNP) gene chip in the Framingham Heart Study. We cross-sectionally evaluated the MetS longitudinal trends. Data analyzed were from the Offspring Cohort (four exams: first (n = 2,441), third (n = 2,185), fifth (n = 2,308), and seventh (n = 2,328)) and the Generation 3 Cohort (one exam: the first exam (n = 3,997)). The prevalence of MetS was determined using the National Cholesterol Education Program Adult Treatment Panel III diagnostic criteria, modified with a newly developed correction for medication use. The association test between an SNP and MetS was performed with a generalized estimating equations method under the additive genetic model. Multiple-testing corrections were also performed. The prevalence of MetS in the offspring cohort increased from one visit to the next, and reached the highest point by the seventh exam comparable with the prevalence reported for the general US population. The pattern of the MetS prevalence over time also reflected itself in the association tests, in which the highest significances were seen in the fifth and seventh exams. The association tests showed that SNPs within genes PRDM16, CETP, PTHB1, PAPPA, and FBN3, and also some SNPs not in genes were significant or close to significance at the genome-wide thresholds. These findings are important in terms of eventually identifying with the causal loci for MetS. PMID- 20017982 TI - Genome-wide association analysis of cardiovascular-related quantitative traits in the Framingham Heart Study. AB - Multivariate linear growth curves were used to model high-density lipoprotein (HDL), low-density lipoprotein (LDL), triglycerides (TG), and systolic blood pressure (SBP) measured during four exams from 1659 independent individuals from the Framingham Heart Study. The slopes and intercepts from each of two phenotype models were tested for association with 348,053 autosomal single-nucleotide polymorphisms from the Affymetrix Gene Chip 500 k set. Three regions were associated with LDL intercept, TG slope, and SBP intercept (p < 1.44 x 10-7). We observed results consistent with previously reported associations between rs599839, on chromosome 1p13, and LDL. We note that the association is significant with LDL intercept but not slope. Markers on chromosome 17q25 were associated with TG slope, and a single-nucleotide polymorphism on chromosome 7p11 was associated with SBP intercept. Growth curve models can be used to gain more insight on the relationships between SNPs and traits than traditional association analysis when longitudinal data has been collected. The power to detect association with changes over time may be limited if the subjects are not followed over a long enough time period. PMID- 20017983 TI - Evaluation of population impact of candidate polymorphisms for coronary heart disease in the Framingham Heart Study Offspring Cohort. AB - In order to evaluate the population impact of putative causal genetic variants over the life course of disease, we extended the static estimation of population attributable risk fraction and developed a novel tool to evaluate how the population impact changes over time using the Framingham Heart Study Offspring Cohort data provided to the Genetic Analysis Workshop 16, Problem 2. A set of population-attributable risk fractions based on survival functions were estimated under the proportional hazards models. The development of this novel measure of population impact creates a more comprehensive estimate of population impact over the life course of disease, which may help us to better understand genetic susceptibility at the population level. PMID- 20017984 TI - A genome-wide association analysis of Framingham Heart Study longitudinal data using multivariate adaptive splines. AB - The Framingham Heart Study is a well known longitudinal cohort study. In recent years, the community-based Framingham Heart Study has embarked on genome-wide association studies. In this paper, we present a Framingham Heart Study genome wide analysis for fasting triglycerides trait in the Genetic Analysis Workshop16 Problem 2 using multivariate adaptive splines for the analysis of longitudinal data (MASAL). With MASAL, we are able to perform analysis of genome-wide data with longitudinal phenotypes and covariates, making it possible to identify genes, gene-gene, and gene-environment (including time) interactions associated with the trait of interest. We conducted a permutation test to assess the associations between MASAL selected markers and triglycerides trait and report significant gene-gene and gene-environment interaction effects on the trait of interest. PMID- 20017985 TI - Adjusting for HLA-DRbeta1 in a genome-wide association analysis of rheumatoid arthritis and related biomarkers. AB - BACKGROUND: There is a long-established association between rheumatoid arthritis and HLA-DRbeta1. The shared epitope (SE) allele is an indicator of the presence of any of the HLA-DRbeta1 alleles associated with RA. Other autoantibodies are also associated with RA, specifically rheumatoid factor IgM (RFUW) and anti cyclic citrullinated peptide (anti-CCP). METHODS: Using the Genetic Analysis Workshop 16 North American Rheumatoid Arthritis Consortium genome-wide association data, we sought to find non-HLA-DRbeta1 genetic associations by stratifying across SE status, and using the continuous biomarker phenotypes of RFUW and anti-CCP. To evaluate the binary RA phenotype, we applied the recently developed FP test and compared it to logistic regression or a genotype count based test. We adjusted for multiple testing using the Bonferroni correction, the Q value approach, or permutation-based p-values. A case-only analysis of the biomarkers RFUW and anti-CCP used linear regression and ANOVAs. RESULTS: The initial genome-wide association analysis using all cases and controls provides substantial evidence of an association on chromosomes 9 and 2 within the immune system-related gene UBXD2. In SE-positive subjects, many single-nucleotide polymorphisms were significant, including some on chromosome 6. Due to very few SE negative cases, we had limited power to detect associations in SE negative subjects. We were also unable to find genetic associations with either RFUW or anti-CCP. CONCLUSION: Our analyses have confirmed previous findings for genes PTPN22 and C5. We also identified a novel candidate gene on chromosome 2, UBXD2. Results suggest FP test may be more powerful than the genotype count-based test. PMID- 20017986 TI - Family-based analysis of a myocardial infarction endophenotype: comparison of sampling designs. AB - The power of linkage analysis of a quantitative disease endophenotype was compared for the following family selection designs: 1) Random samples: randomly chosen nuclear families, 2) "coronary artery calcification (CAC)" samples: selection of each nuclear family through a proband with abnormally high levels of the simulated quantitative endophenotype, CAC, and (3) "MI" samples: selection of each nuclear family through a disease affected proband, in this case a proband who had been simulated to have a myocardial infarction (MI) event.We assessed the power to detect linkage to five loci (two pairs of epistatic loci and one locus with an over-dominant allele) that were modeled as determinants of the simulated CAC levels. We did this using a Haseman-Elston regression-based linkage analysis of the adjusted CAC levels that considered each locus separately and then used a multiple regression extension of the Haseman-Elston method in which we considered the allele sharing at two true epistatic loci simultaneously and their interaction as possible factors related to the squared sibpair differences in adjusted CAC.Based on comparison of the mean square root of the LOD scores, there was no one sampling design that resulted in consistently greater power for these five loci. That is, we observed significant locus-by-sampling-design interaction (p < 0.0001). We noted however, that the largest average score was observed for the epistasis between tau3 and tau4 (mean > 1.8, SE = 0.06) in the MI-selected samples and the CAC-selected samples. PMID- 20017987 TI - Pedigree association: assigning individual weights to pedigree members for genetic association analysis. AB - OBJECTIVE: Methods exist to appropriately perform association analyses in pedigrees. However, for genome-wide association analysis, these methods are computationally impractical. It is therefore important to determine alternate methods that can be efficiently used genome-wide. Here, we introduce a new algorithm that considers all relationships simultaneously in arbitrary-structured pedigrees and assigns weights to pedigree members that can be used in subsequent analyses to address relatedness. We compare this new method with an existing weighting algorithm, a naive analysis (relatedness is ignored), and an empirical method that appropriately accounts for all relationships (the gold standard). METHODS: Framingham Heart Study Genetic Analysis Workshop 16 Problem 2 data were used with a dichotomous phenotype based on high-density lipoprotein cholesterol level (1,611 cases and 4,043 controls). New and existing algorithms for calculating weights were used. Cochran-Armitage trend tests were performed for 17,333 single-nucleotide polymorphisms on chromosome 8 using both weighting systems and the naive approach; a subset of 500 single-nucleotide polymorphisms were tested empirically. Correlations of p-values from each method were determined. RESULTS: Results from the two weighting methods were strongly correlated (r = 0.96). Our new weighting method performed better than the existing weighting method (r = 0.89 vs. r = 0.83), which is due to a more moderate down-weighting. The naive analysis obtained the best correlation with the empirical gold standard results (r = 0.99). CONCLUSION: Our results suggest that weighting methods do not accurately represent tests that account for familial relationships in genetic association analyses and are inferior to the naive method as an efficient initial genome-wide screening tool. PMID- 20017988 TI - A comparative study of three methods for detecting association of quantitative traits in samples of related subjects. AB - We used Genetic Analysis Workshop 16 Problem 3 Framingham Heart Study simulated data set to compare methods for association analysis of quantitative traits in related individuals. More specifically, we investigated type I error and relative power of three approaches: the measured genotype, the quantitative transmission disequilibrium test (QTDT), and the quantitative trait linkage-disequilibrium (QTLD) tests. We studied high-density lipoprotein and triglyceride (TG) lipid variables, as measured at Visit 1. Knowing the answers, we selected three true major genes for high-density lipoprotein and/or TG. Empirical distributions of the three association models were derived from the first 100 replicates. In these data, all three models were similar in error rates. Across the three association models, the power was the lowest for the functional SNP with smallest size effects (i.e., alpha2), and for the less heritable trait (i.e., TG). Our results showed that measured genotype outperformed the two orthogonal-based association models (QTLD, QTDT), even after accounting for population stratification. QTDT had the lowest power rates. This is consistent with the amount of marker and trait data used by each association model. While the effective sample sizes varied little across our tested variants, we observed some large power drops and marked differences in performances of the models. We found that the performances contrasted the most for the tightly linked, but not associated, functional variants. PMID- 20017989 TI - Testing for genetic association taking into account phenotypic information of relatives. AB - We investigated efficient case-control association analysis using family data. The outcome of interest was coronary heart disease. We employed existing and new methods that take into account the correlations among related individuals to obtain the proper type I error rates. The methods considered for autosomal single nucleotide polymorphisms were: 1) generalized estimating equations-based methods, 2) variance-modified Cochran-Armitage (MCA) trend test incorporating kinship coefficients, and 3) genotypic modified quasi-likelihood score test. Additionally, for X-linked single-nucleotide polymorphisms we proposed a two degrees-of-freedom test. Performance of these methods was tested using Framingham Heart Study 500 k array data. PMID- 20017990 TI - Family-based genome-wide association study for simulated data of Framingham Heart Study. AB - Genome-wide association studies (GWAS) have quickly become the norm in dissecting the genetic basis of complex diseases. Family-based association approaches have the advantages of being robust to possible hidden population structure in samples. Most of these methods were developed with limited markers. Their applicability and performance for GWAS need to be examined. In this report, we evaluated the properties of the family-based association method implemented by ASSOC in the S.A.G.E package using the simulated data sets for the Framingham Heart Study, and found that ASSOC is a highly useful tool for GWAS. PMID- 20017991 TI - Detection of imprinting and heterogeneous maternal effects on high blood pressure using Framingham Heart Study data. AB - Both imprinting and maternal effects could lead to parent-of-origin patterns in complex traits of human disorders. Statistical methods that differentiate these two effects and identify them simultaneously by using family-based data from retrospective studies are available. The usual data structures include case parents triads and nuclear families with multiple affected siblings. We develop a likelihood-based method to detect imprinting and maternal effects simultaneously using data from prospective studies. The proposed method utilizes both affected and unaffected siblings in nuclear families by modeling familial genotypes and offspring's disease status jointly. Maternal effect is usually modeled as a fixed effect under the assumption that maternal variant allele(s) has (have) identical effect on any offspring. However, recent studies report that different people may carry different amounts of substances encoded by the mother's variant allele(s) (called maternal microchimerism), which could result in heterogeneity of maternal effects. The proposed method incorporates the heterogeneity of maternal effects by adding a random component to the logit of the penetrance. Our method was applied to the Framingham Heart Study data in two steps to detect single nucleotide polymorphisms (SNPs) that may be associated with high blood pressure. In the first step, SNPs that affect susceptibility of high blood pressure through minor allele, genomic imprinting, or maternal effects were identified by using the proposed model without the random effect component. In the second step, we fitted the mixed effect model to the identified SNPs that have significant maternal effect to detect heterogeneity of the maternal effects. PMID- 20017992 TI - Gene hunting of the Genetic Analysis Workshop 16 rheumatoid arthritis data using rough set theory. AB - We propose to use the rough set theory to identify genes affecting rheumatoid arthritis risk from the data collected by the North American Rheumatoid Arthritis Consortium. For each gene, we employ generalized dynamic reducts in the rough set theory to select a subset of single-nucleotide polymorphisms (SNPs) to represent the genetic information from this gene. We then group the study subjects into different clusters based on their genotype similarity at the selected markers. Statistical association between disease status and cluster membership is then studied to identify genes associated with rheumatoid arthritis. Based on our proposed approach, we are able to identify a number of statistically significant genes associated with rheumatoid arthritis. Aside from genes on chromosome 6, our identified genes include known disease-associated genes such as PTPN22 and TRAF1. In addition, our list contains other biologically plausible genes, such as ADAM15 and AGPAT2. Our findings suggest that ADAM15 and AGPAT2 may contribute to a genetic predisposition through abnormal angiogenesis and adipose tissue. PMID- 20017993 TI - Region-based analysis in genome-wide association study of Framingham Heart Study blood lipid phenotypes. AB - Due to the high-dimensionality of single-nucleotide polymorphism (SNP) data, region-based methods are an attractive approach to the identification of genetic variation associated with a certain phenotype. A common approach to defining regions is to identify the most significant SNPs from a single-SNP association analysis, and then use a gene database to obtain a list of genes proximal to the identified SNPs. Alternatively, regions may be defined statistically, via a scan statistic. After categorizing SNPs as significant or not (based on the single-SNP association p-values), a scan statistic is useful to identify regions that contain more significant SNPs than expected by chance. Important features of this method are that regions are defined statistically, so that there is no dependence on a gene database, and both gene and inter-gene regions can be detected. In the analysis of blood-lipid phenotypes from the Framingham Heart Study (FHS), we compared statistically defined regions with those formed from the top single SNP tests. Although we missed a number of single SNPs, we also identified many additional regions not found as SNP-database regions and avoided issues related to region definition. In addition, analyses of candidate genes for high-density lipoprotein, low-density lipoprotein, and triglyceride levels suggested that associations detected with region-based statistics are also found using the scan statistic approach. PMID- 20017994 TI - Pathway-based analysis of a genome-wide case-control association study of rheumatoid arthritis. AB - Evaluation of the association between single-nucleotide polymorphisms (SNPs) and disease outcomes is widely used to identify genetic risk factors for complex diseases. Although this analysis paradigm has made significant progress in many genetic studies, many challenges remain, such as the requirement of a large sample size to achieve adequate power. Here we use rheumatoid arthritis (RA) as an example and explore a new analysis strategy: pathway-based analysis to search for related genes and SNPs contributing to the disease.We first propose the application of measure of explained variation to quantify the predictive ability of a given SNP. We then use gene set enrichment analysis to evaluate enrichment of specific pathways, where pathways, are considered enriched if they consist of genes that are associated with the phenotype of interest above and beyond is expected by chance. The results are also compared with score tests for association analysis by adjusting for population stratification.Our study identified some significantly enriched pathways, such as "cell adhesion molecules," which are known to play a key role in RA. Our results showed that pathway-based analysis may identify other biologically interesting loci (e.g., rs1018361) related to RA: the gene (CTLA4) closest to this marker has previously been shown to be associated with RA and the gene is in the significant pathways we identified, even though the marker has not reached genome-wide significance in univariate single-marker analysis. PMID- 20017995 TI - A principal-components-based clustering method to identify multiple variants associated with rheumatoid arthritis and arthritis-related autoantibodies. AB - Multivariate techniques are an important area of investigation for studying contributions of multiple genetic variants to disease onset and pathology. We analyzed the Genetic Analysis Workshop 16 North American Rheumatoid Arthritis Consortium (NARAC) data using a principal-components analysis (PCA) with an orthoblique rotation to identify specific subsets of single-nucleotide polymorphisms (SNP) in the major histocompatibility complex (MHC) region associated with rheumatoid arthritis (RA) and rheumatoid factor IgM (RFUW), and compared this method with a traditional PC approach. Using the orthoblique PC based clustering method, we identified new clusters of SNPs across the MHC region associated with RA and RFUW, and replicated known SNP cluster associations with RA, such as those in the HLA-DRB region. PMID- 20017996 TI - Effect of population stratification on the identification of significant single nucleotide polymorphisms in genome-wide association studies. AB - The North American Rheumatoid Arthritis Consortium case-control study collected case participants across the United States and control participants from New York. More than 500,000 single-nucleotide polymorphisms (SNPs) were genotyped in the sample of 2000 cases and controls. Careful adjustment for the confounding effect of population stratification must be conducted when analyzing these data; the variance inflation factor (VIF) without adjustment is 1.44. In the primary analyses of these data, a clustering algorithm in the program PLINK was used to reduce the VIF to 1.14, after which genomic control was used to control residual confounding. Here we use stratification scores to achieve a unified and coherent control for confounding. We used the first 10 principal components, calculated genome-wide using a set of 81,500 loci that had been selected to have low pair wise linkage disequilibrium, as risk factors in a logistic model to calculate the stratification score. We then divided the data into five strata based on quantiles of the stratification score. The VIF of these stratified data is 1.04, indicating substantial control of stratification. However, after control for stratification, we find that there are no significant loci associated with rheumatoid arthritis outside of the HLA region. In particular, we find no evidence for association of TRAF1-C5 with rheumatoid arthritis. PMID- 20017997 TI - A new gene-based association test for genome-wide association studies. AB - Genome-wide association studies are widely used today to discover genetic factors that modify the risk of complex diseases. Usually, these methods work in a SNP-by-SNP fashion. We present a gene-based test that can be applied in the context of genome-wide association studies. We compare both strategies, SNP-based and gene-based, in a sample of cases and controls for rheumatoid arthritis.We obtained different results using each strategy. The SNP-based test found the PTPN22 gene while the gene-based test found the PHF19-TRAF1-C5 region. That suggests that no single strategy performs better than another in all cases and that a certain underlying genetic architecture can be delineated more easily with one strategy rather than with another. PMID- 20017998 TI - Identification of novel putative rheumatoid arthritis susceptibility genes via analysis of rare variants. AB - Established loci for rheumatoid arthritis (RA), including HLA-DRB1 and PTPN22, do not fully account for the genetic component of susceptibility to the disease. One possible source of as yet undiscovered susceptibility genes are those mediated through effects of rare variants. We present a novel method for gene based genome-wide scans of whole-genome association (WGA) data to identify accumulations of rare variants associated with disease. We apply our method to WGA SNP genotype data obtained from 868 RA cases and 1194 controls. Our results highlight novel putative RA susceptibility genes that have not previously been identified in large-scale WGA studies. PMID- 20017999 TI - Genome-wide gene-based analysis of rheumatoid arthritis-associated interaction with PTPN22 and HLA-DRB1. AB - The genes PTPN22 and HLA-DRB1 have been found by a number of studies to confer an increased risk for rheumatoid arthritis (RA), which indicates that both genes play an important role in RA etiology. It is believed that they not only have strong association with RA individually, but also interact with other related genes that have not been found to have predisposing RA mutations. In this paper, we conduct genome-wide searches for RA-associated gene-gene interactions that involve PTPN22 or HLA-DRB1 using the Genetic Analysis Workshop 16 Problem 1 data from the North American Rheumatoid Arthritis Consortium. MGC13017, HSPCAL3, MIA, PTPNS1L, and IGLVI-70, which showed association with RA in previous studies, have been confirmed in our analysis. PMID- 20018000 TI - Copy number variation in the Framingham Heart Study. AB - In this paper we test for association between copy number variation and diabetes in a subset of individuals from the Framingham Heart Study. We used the 500 k SNP data and called copy number variation using two algorithms: the genome alteration detection algorithm of Pique-Regi et al. and the software Golden Helix. We then tested for association between copy number and diabetes using a gene-based analysis. Our results show little evidence of association between copy number and diabetes status. Furthermore, our results indicate a relatively poor level of agreement between copy number calls resulting from the two programs. We then examined potential causes for this difference in results and the implications for future studies. PMID- 20018001 TI - Association of KCNB1 to rheumatoid arthritis via interaction with HLA-DRB1. AB - With the rapid development of large-scale high-throughput genotyping technology, genome-wide association studies have become a popular approach to mapping genes underlying common human disorders. Some genes are discovered, but many more have not been. Because these genes were not initially identified, it is reasonable to assume that their main effect is weak. We propose a method to accommodate such a situation. It is applied to the Genetic Analysis Workshop 16 Problem 1 case control data in which shared-epitope alleles of HLA-DRB1 show very strong association with rheumatoid arthritis. Because some previous functional studies have reported association of gene KCNB1 to rheumatoid arthritis, we evaluate whether the gene KCNB1 contributes to the genetics of rheumatoid arthritis in this data set. Fifteen single-nucleotide polymorphisms from this gene were chosen. The association of KCNB1 gene to rheumatoid arthritis seems to be moderate. PMID- 20018002 TI - Genome-wide gene-based association study. AB - Genome-wide association studies, which analyzes hundreds of thousands of single nucleotide polymorphisms to identify disease susceptibility genes, are challenging because the work involves intensive computation and complex modeling. We propose a two-stage genome-wide association scanning procedure, consisting of a single-locus association scan for the first stage and a gene-based association scan for the second stage. Marginal effects of single-nucleotide polymorphisms are examined by using the exact Armitage trend test or logistic regression, and gene effects are examined by using a p-value combination method. Compared with some existing single-locus and multilocus methods, the proposed method has the following merits: 1) convenient for definition of biologically meaningful regions, 2) powerful for detection of minor-effect genes, 3) helpful for alleviation of a multiple-testing problem, and 4) convenient for result interpretation. The method was applied to study Genetic Analysis Workshop 16 Problem 1 rheumatoid arthritis data, and strong association signals were found. The results show that the human major histocompatibility complex region is the most important genomic region associated with rheumatoid arthritis. Moreover, previously reported genes including PTPN22, C5, and IL2RB were confirmed; novel genes including HLA-DRA, BTNL2, C6orf10, NOTCH4, TAP2, and TNXB were identified by our analysis. PMID- 20018003 TI - Detecting susceptibility genes for rheumatoid arthritis based on a novel sliding window approach. AB - With the recent rapid improvements in high-throughout genotyping techniques, researchers are facing a very challenging task of large-scale genetic association analysis, especially at the whole-genome level, without an optimal solution. In this study, we propose a new approach for genetic association analysis based on a variable-sized sliding-window framework. This approach employs principal component analysis to find the optimal window size. Using the bisection algorithm in window size searching, the proposed method tackles the exhaustive computation problem. It is more efficient and effective than currently available approaches. We conduct the genome-wide association study in Genetic Analysis Workshop 16 (GAW16) Problem 1 data using the proposed method. Our method successfully identified several susceptibility genes that have been reported by other researchers and additional candidate genes for follow-up studies. PMID- 20018004 TI - A search for non-chromosome 6 susceptibility loci contributing to rheumatoid arthritis. AB - We conducted a search for non-chromosome 6 genes that may increase risk for rheumatoid arthritis (RA). Our approach was to retrospectively ascertain three "extreme" subsamples from the North American Rheumatoid Arthritis Consortium. The three subsamples are: 1) RA cases who have two low-risk HLA-DRB1 alleles (N = 18), 2) RA cases who have two high-risk HLA-DRB1 alleles (N = 163), and 3) controls who have two low-risk HLA-DRB1 alleles (N = 652). We hypothesized that since Group 1's RA was likely due to non-HLA related risk factors, and because Group 3, by definition, is unaffected, comparing Group 1 with Group 2 and Group 1 with Group 3 would result in the identification of candidate susceptibility loci located outside of the MHC region. Accordingly, we restricted our search to the 21 non-chromosome 6 autosomes. The case-case comparison of Groups 1 and 2 resulted in the identification of 17 SNPs with allele frequencies that differed at p < 0.0001. The case-control comparison of Groups 1 and 3 identified 23 SNPs that differed in allele frequency at p < 0.0001. Eight of these SNPs (rs10498105, rs2398966, rs7664880, rs7447161, rs2793471, rs2611279, rs7967594, and rs742605) were common to both lists. PMID- 20018005 TI - Analysis of genome-wide association data by large-scale Bayesian logistic regression. AB - Single-locus analysis is often used to analyze genome-wide association (GWA) data, but such analysis is subject to severe multiple comparisons adjustment. Multivariate logistic regression is proposed to fit a multi-locus model for case control data. However, when the sample size is much smaller than the number of single-nucleotide polymorphisms (SNPs) or when correlation among SNPs is high, traditional multivariate logistic regression breaks down. To accommodate the scale of data from a GWA while controlling for collinearity and overfitting in a high dimensional predictor space, we propose a variable selection procedure using Bayesian logistic regression. We explored a connection between Bayesian regression with certain priors and L1 and L2 penalized logistic regression. After analyzing large number of SNPs simultaneously in a Bayesian regression, we selected important SNPs for further consideration. With much fewer SNPs of interest, problems of multiple comparisons and collinearity are less severe. We conducted simulation studies to examine probability of correctly selecting disease contributing SNPs and applied developed methods to analyze Genetic Analysis Workshop 16 North American Rheumatoid Arthritis Consortium data. PMID- 20018006 TI - Case-control genome-wide association study of rheumatoid arthritis from Genetic Analysis Workshop 16 using penalized orthogonal-components regression-linear discriminant analysis. AB - Currently, genome-wide association studies (GWAS) are conducted by collecting a massive number of SNPs (i.e., large p) for a relatively small number of individuals (i.e., small n) and associations are made between clinical phenotypes and genetic variation one single-nucleotide polymorphism (SNP) at a time. Univariate association approaches like this ignore the linkage disequilibrium between SNPs in regions of low recombination. This results in a low reliability of candidate gene identification. Here we propose to improve the case-control GWAS approach by implementing linear discriminant analysis (LDA) through a penalized orthogonal-components regression (POCRE), a newly developed variable selection method for large p small n data. The proposed POCRE-LDA method was applied to the Genetic Analysis Workshop 16 case-control data for rheumatoid arthritis (RA). In addition to the two regions on chromosomes 6 and 9 previously associated with RA by GWAS, we identified SNPs on chromosomes 10 and 18 as potential candidates for further investigation. PMID- 20018007 TI - A quantile-based method for association mapping of quantitative phenotypes: an application to rheumatoid arthritis phenotypes. AB - Genetic association of population-based quantitative trait data has traditionally been analyzed using analysis of variance (ANOVA). However, violations of certain statistical assumptions may lead to false-positive association results. In this study, we have explored model-free alternatives to ANOVA using correlations between allele frequencies in the different quantile intervals of the quantitative trait and the quantile values. We performed genome wide association scans on anti-cyclic citrullinated peptide and rheumatoid factor immunoglobulin M, two quantitative traits correlated with rheumatoid arthritis, using the data provided in Genetic Analysis Workshop 16. Both the quantitative traits exhibited significant evidence of association on Chromosome 6, although not in the human leukocyte antigen region which is known to harbor a major gene predisposing to rheumatoid arthritis. We found that while a majority of the significant findings using the asymptotic thresholds of ANOVA was not validated using permutations, a relatively higher proportion of the significant findings using the asymptotic cut-offs of the correlation statistic were validated using permutations. PMID- 20018008 TI - Genome-wide discovery of maternal effect variants. AB - Many phenotypes may be influenced by the prenatal environment of the mother and/or maternal care, and these maternal effects may have a heritable component. We have implemented in the computer program SOLAR a variance components-based method for detecting indirect effects of maternal genotype on offspring phenotype. Of six phenotypes measured in three generations of the Framingham Heart Study, height showed the strongest evidence (P = 0.02) of maternal effect. We conducted a genome-wide association analysis for height, testing both the direct effect of the focal individual's genotype and the indirect effect of the maternal genotype. Offspring height showed suggestive evidence of association with maternal genotype for two single-nucleotide polymorphisms in the trafficking protein particle complex 9 gene TRAPPC9 (NIBP), which plays a role in neuronal NF kappaB signalling. This work establishes a methodological framework for identifying genetic variants that may influence the contribution of the maternal environment to offspring phenotypes. PMID- 20018009 TI - Data for Genetic Analysis Workshop 16 Problem 1, association analysis of rheumatoid arthritis data. AB - For Genetic Analysis Workshop 16 Problem 1, we provided data for genome-wide association analysis of rheumatoid arthritis. Single-nucleotide polymorphism (SNP) genotype data were provided for 868 cases and 1194 controls that had been assayed using an Illumina 550 k platform. In addition, phenotypic data were provided from genotyping DRB1 alleles, which were classified according to the rheumatoid arthritis shared epitope, levels of anti-cyclic citrullinated peptide, and levels of rheumatoid factor IgM. Several questions could be addressed using the data, including analysis of genetic associations using single SNPs or haplotypes, as well as gene-gene and genetic analysis of SNPs for qualitative and quantitative factors. PMID- 20018010 TI - Simultaneous genome-wide association studies of anti-cyclic citrullinated peptide in rheumatoid arthritis using penalized orthogonal-components regression. AB - Genome-wide associations between single-nucleotide polymorphisms and clinical traits were simultaneously conducted using penalized orthogonal-components regression. This method was developed to identify the genetic variants controlling phenotypes from a massive number of candidate variants. By investigating the association between all single-nucleotide polymorphisms to the phenotype of antibodies against cyclic citrullinated peptide using the rheumatoid arthritis data provided by Genetic Analysis Workshop 16, we identified genetic regions which may contribute to the pathogenesis of rheumatoid arthritis. Bioinformatic analysis of these genomic regions showed most of them harbor protein-coding gene(s). PMID- 20018011 TI - Comparison between the stochastic search variable selection and the least absolute shrinkage and selection operator for genome-wide association studies of rheumatoid arthritis. AB - BACKGROUND: Because multiple loci control complex diseases, there is great interest in testing markers simultaneously instead of one by one. In this paper, we applied two model selection algorithms: the stochastic search variable selection (SSVS) and the least absolute shrinkage and selection operator (LASSO) to two quantitative phenotypes related to rheumatoid arthritis (RA). RESULTS: The Genetic Analysis Workshop 16 data includes 2,062 unrelated individuals and 545,080 single-nucleotide polymorphism markers from the Illumina 550 k chip. We performed our analyses on the cases as the quantitative phenotype data was not provided for the controls. The performance of the two algorithms was compared. Using sure independence screening as the prescreening procedure, both SSVS and LASSO give small models. No markers are identified in the human leukocyte antigen region of chromosome 6 that was shown to be associated with RA. SSVS and LASSO identify seven common loci, and some of them are on genes LRRC8D, LRP1B, and COLEC12. These genes have not been reported to be associated with RA. LASSO also identified a common locus on gene KTCD21 for the two phenotypes (marker rs230662 and rs483731, respectively). CONCLUSION: SSVS outperforms LASSO in simulation studies. Both SSVS and LASSO give small models on the RA data, however this depends on model parameters. We also demonstrate the ability of both LASSO and SSVS to handle more markers than the number of samples. PMID- 20018012 TI - Comparison of a unified analysis approach for family and unrelated samples with the transmission-disequilibrium test to study associations of hypertension in the Framingham Heart Study. AB - Population stratification is one of the major causes of spurious associations in association studies. A unified association approach based on principal-component analysis can overcome the effect of population stratification, as well as make use of both family and unrelated samples combined to increase power (family-case control, or FamCC). In this study, we compared FamCC and the transmission disequilibrium test (TDT) using data on hypertension, systolic blood pressure, and diastolic blood pressure in the Framingham Heart Study. Our study indicated FamCC has reasonable type I error for both the unrelated sample and the family sample for all three traits. For these three traits, we found results from FamCC were inconsistent with those from the TDT. We discuss the reasons for this inconsistency. After correcting for multiple tests, we did not detect any significant single-nucleotide polymorphisms by either FamCC or the TDT. PMID- 20018013 TI - Power of selective genotyping in genome-wide association studies of quantitative traits. AB - The selective genotyping approach in quantitative genetics means genotyping only individuals with extreme phenotypes. This approach is considered an efficient way to perform gene mapping, and can be applied in both linkage and association studies. Selective genotyping in association mapping of quantitative trait loci was proposed to increase the power of detecting rare alleles of large effect. However, using this approach, only common variants have been detected. Studies on selective genotyping have been limited to single-locus scenarios. In this study we aim to investigate the power of selective genotyping in a genome-wide association study scenario, and we specifically study the impact of minor allele frequency of variants on the power of this approach. We use the Genetic Analysis Workshop 16 rheumatoid arthritis whole-genome data from the North American Rheumatoid Arthritis Consortium. Two quantitative traits, anti-cyclic citrullinated peptide and rheumatoid factor immunoglobulin M, and one binary trait, rheumatoid arthritis affection status, are used in the analysis. The power of selective genotyping is explored as a function of three parameters: sampling proportion, minor allele frequency of single-nucleotide polymorphism, and test level. The results show that the selective genotyping approach is more efficient in detecting common variants than detecting rare variants, and it is efficient only when the level of declaring significance is not stringent. In summary, the selective genotyping approach is most suitable for detecting common variants in candidate gene-based studies. PMID- 20018014 TI - Application of imputation methods to the analysis of rheumatoid arthritis data in genome-wide association studies. AB - Most genetic association studies only genotype a small proportion of cataloged single-nucleotide polymorphisms (SNPs) in regions of interest. With the catalogs of high-density SNP data available (e.g., HapMap) to researchers today, it has become possible to impute genotypes at untyped SNPs. This in turn allows us to test those untyped SNPs, the motivation being to increase power in association studies. Several imputation methods and corresponding software packages have been developed for this purpose. The objective of our study is to apply three widely used imputation methods and corresponding software packages to a data from a genome-wide association study of rheumatoid arthritis from the North American Rheumatoid Arthritis Consortium in Genetic Analysis Workshop 16, to compare the performances of the three methods, to evaluate their strengths and weaknesses, and to identify additional susceptibility loci underlying rheumatoid arthritis. The software packages used in this paper included a program for Bayesian imputation-based association mapping (BIMBAM), a program for imputing unobserved genotypes in case-control association studies (IMPUTE), and a program for testing untyped alleles (TUNA). We found some untyped SNP that showed significant association with rheumatoid arthritis. Among them, a few of these were not located near any typed SNP that was found to be significant and thus may be worth further investigation. PMID- 20018015 TI - Elastic-net regularization approaches for genome-wide association studies of rheumatoid arthritis. AB - The current trend in genome-wide association studies is to identify regions where the true disease-causing genes may lie by evaluating thousands of single nucleotide polymorphisms (SNPs) across the whole genome. However, many challenges exist in detecting disease-causing genes among the thousands of SNPs. Examples include multicollinearity and multiple testing issues, especially when a large number of correlated SNPs are simultaneously tested. Multicollinearity can often occur when predictor variables in a multiple regression model are highly correlated, and can cause imprecise estimation of association. In this study, we propose a simple stepwise procedure that identifies disease-causing SNPs simultaneously by employing elastic-net regularization, a variable selection method that allows one to address multicollinearity. At Step 1, the single-marker association analysis was conducted to screen SNPs. At Step 2, the multiple-marker association was scanned based on the elastic-net regularization. The proposed approach was applied to the rheumatoid arthritis (RA) case-control data set of Genetic Analysis Workshop 16. While the selected SNPs at the screening step are located mostly on chromosome 6, the elastic-net approach identified putative RA related SNPs on other chromosomes in an increased proportion. For some of those putative RA-related SNPs, we identified the interactions with sex, a well known factor affecting RA susceptibility. PMID- 20018016 TI - Application of seventeen two-locus models in genome-wide association studies by two-stage strategy. AB - The goal of this paper is to search for two-locus combinations that are jointly associated with rheumatoid arthritis using the data set of Genetic Analysis Workshop 16 Problem 1. We use a two-stage strategy to reduce the computational burden associated with performing an exhaustive two-locus search across the genome. In the first stage, the full set of 531,689 single-nucleotide polymorphisms was screened using univariate testing. In the second stage, all pairs made from the 500 single-nucleotide polymorphisms with the lowest p-values from the first stage were evaluated under each of 17 two-locus models. Our analyses identified a two-locus combination - rs6939589 and rs11634386 - that proved to be significantly associated with rheumatoid arthritis under a Rec x Rec model (p-value = 0.045 after adjusting for multiple tests and multiple models). PMID- 20018017 TI - Comparison between single-marker analysis using Merlin and multi-marker analysis using LASSO for Framingham simulated data. AB - We compared family-based single-marker association analysis using Merlin and multi-marker analysis using LASSO (least absolute shrinkage and selection operator) for the low-density lipoprotein phenotype at the first visit for all 200 replicates of the Genetic Analysis Workshop 16 Framingham simulated data sets. Using "answers," we selected single-nucleotide polymorphisms (SNPs) on chromosome 22 for comparison of results between single-marker and multi-marker analyses. For the major causal SNP rs2294207 on chromosome 22, both single-marker and multi-marker analyses provided similar results, indicating the importance of this SNP. For the 12 polygenic SNPs on the same chromosome, both single-marker and multi-marker analyses failed to provide statistically significant associations, indicating that their effects were too weak to be detected by either method. The main difference between the two methods was that for the 14 SNPs near the causal SNPs, p-values from Merlin were the next smallest, whereas LASSO often excluded these non-causal neighboring SNPs entirely from the first 10,000 models. PMID- 20018018 TI - Incorporating multiple-marker information to detect risk loci for rheumatoid arthritis. AB - In genome-wide association studies, new schemes are needed to incorporate multiple-locus information. In this article, we proposed a two-stage sliding window approach to detect associations between a disease and multiple genetic polymorphisms. In the proposed approach, we measured the genetic association between a disease and a single-nucleotide polymorphism window by the newly developed likelihood ratio test-principal components statistic, and performed a sliding-window technique to detect disease susceptibility windows. We split the whole sample into two sub-samples, each of which contained a portion of cases and controls. In the first stage, we selected the top R windows by the statistics based on the first sub-sample, and in the second stage, we claimed significant windows by false-discovery rate correction on the p-values of the statistics based on the second sub-sample. By applying the new approach to the Genetic Analysis Workshop 16 Problem 1 data set, we detected 212 out of 531,601 windows to be responsible for rheumatoid arthritis. Except for chromosomes 4 and 18, each of the other 20 autosomes was found to harbor risk windows. Our results supported the findings of some rheumatoid arthritis susceptibility genes identified in the literature. In addition, we identified several new single-nucleotide polymorphism windows for follow-up studies. PMID- 20018019 TI - Two-stage joint selection method to identify candidate markers from genome-wide association studies. AB - The interaction among multiple genes and environmental factors can affect an individual's susceptibility to disease. Some genes may not show strong marginal associations when they affect disease risk through interactions with other genes. As a result, these genes may not be identified by single-marker methods that are widely used in genome-wide association studies. To explore this possibility in real data, we carried out a two-stage model selection procedure of joint single nucleotide polymorphism (SNP) analysis to detect genes associated with rheumatoid arthritis (RA) using Genetic Analysis Workshop 16 genome-wide association study data. In the first stage, the genetic markers were screened through an exhaustive two-dimensional search, through which promising SNP and SNP pairs were identified. Then, LASSO was used to choose putative SNPs from the candidates identified in the first stage. We then use the RA data collected by the Wellcome Trust Case Control Consortium to validate the putative genetic factors. Balancing computational load and statistical power, this method detects joint effects that may fail to emerge from single-marker analysis. Based on our proposed approach, we not only replicated the identification of important RA risk genes, but also found novel genes and their epistatic effects on RA. To our knowledge, this is the first two-dimensional scan based analysis for a real genome-wide association study. PMID- 20018020 TI - Genetics Analysis Workshop 16 Problem 2: the Framingham Heart Study data. AB - Genetic Analysis Workshop 16 (GAW16) Problem 2 presented data from the Framingham Heart Study (FHS), an observational, prospective study of risk factors for cardiovascular disease begun in 1948. Data have been collected in three generations of family participants in the study and the data presented for GAW16 included phenotype data from all three generations, with four examinations of data collected repeatedly for the first two generations. The trait data consisted of information on blood pressure, hypertension treatment, lipid levels, diabetes and blood glucose, smoking, alcohol consumed, weight, and coronary heart disease incidence. Additionally, genotype data obtained through a genome-wide scan (FHS SHARe) of 550,000 single-nucleotide polymorphisms from Affymetrix chips were included with the GAW16 data. The genotype data were also used for GAW16 Problem 3, where simulated phenotypes were generated using the actual FHS genotypes. These data served to provide investigators with a rich resource to study the behavior of genome-wide scans with longitudinally collected family data and to develop and apply new procedures. PMID- 20018021 TI - Genome-wide association analysis of rheumatoid arthritis data via haplotype sharing. AB - We present computationally simple association tests based on haplotype sharing that can be easily applied to genome-wide association studies, while allowing use of fast (but not likelihood-based) haplotyping algorithms, and properly accounting for the uncertainty introduced by using inferred haplotypes. We also give haplotype sharing analyses that adjust for population stratification. We apply our methods to a genome-wide association study of rheumatoid arthritis available as Problem 1 of Genetic Analysis Workshop 16. In addition to the HLA region on chromosome 6, we find genome-wide significant signals at 7q33 and 13q31.3. These regions contain genes with interesting potential connections with rheumatoid arthritis and are not identified using single single-nucleotide polymorphism methods. PMID- 20018022 TI - Replication of recently identified associated single-nucleotide polymorphisms from six autoimmune diseases in Genetic Analysis Workshop 16 rheumatoid arthritis data. AB - Many autoimmune diseases share similar underlying pathology and have a tendency to cluster within families, giving rise to the concept of shared susceptibility genes among them. In the Genetic Analysis Workshop 16 rheumatoid arthritis (RA) data we sought to replicate the genetic association between single-nucleotide polymorphisms (SNPs) identified in recent genome-wide association studies (GWAS) on RA and five other autoimmune diseases. We identified 164 significantly associated non-HLA SNPs (p < 10-5) from 16 GWAS and 13 candidate gene studies on six different autoimmune diseases, including RA, systemic lupus erythematosus, type 1 diabetes, Crohn disease, multiple sclerosis, and celiac disease. Using both direct and imputation-based association test, we replicated 16 shared susceptibility regions involving RA and at least one of the other autoimmune diseases. We also identified hidden population structure within cases and controls in Genetic Analysis Workshop 16 RA data and assessed the effect of population structure on the shared autoimmunity regions. Because multiple autoimmune diseases share common genetic origin, these could be areas of immense interest for further genetic and clinical association studies. PMID- 20018023 TI - Haplotype association analysis of North American Rheumatoid Arthritis Consortium data using a generalized linear model with regularization. AB - The Genetic Analysis Workshop 16 rheumatoid arthritis data include a set of 868 cases and 1194 controls genotyped at 545,080 single-nucleotide polymorphisms (SNPs) from the Illumina 550 k chip. We focus on investigating chromosomes 6 and 18, which have 35,574 and 16,450 SNPs, respectively. Association studies, including single SNP and haplotype-based analyses, were applied to the data on those two chromosomes. Specifically, we conducted a generalized linear model with regularization (rGLM) approach for detecting disease-haplotype association using unphased SNP data. A total of 444 and 43 four-SNP tests were found to be significant at the Bonferroni corrected 5% significance level on chromosome 6 and 18, respectively. PMID- 20018024 TI - On the association between rheumatoid arthritis and classical HLA class I and class II alleles predicted from single-nucleotide polymorphism data. AB - Using single-nucleotide polymorphisms (SNPs), we sought to predict classical class I and class II human leukocyte antigen (HLA) alleles, and test for their associations with rheumatoid arthritis (RA) in the North American Rheumatoid Arthritis Consortium sample of cases and controls, genotyped on the Illumina HumanHap550 BeadChip. We use publicly available databases of SNP data and HLA data to find SNPs or SNP-haplotypes to be used as surrogates for each HLA allele. To reduce the confounding effects of linkage disequilibrium with the HLA-DRB1 locus, we tested for the association conditional on the presence or absence of a shared epitope allele on the same haplotype as the target HLA allele. Using SNP surrogates, we find that components of the DQ8 serotype (DQA1*0301:DQB1*0302) are associated with RA, irrespective of the presence or absence of a shared epitope allele on their respective haplotypes. Knowledge of the haplotype structure in the HLA region is still necessary for better interpretation of the results. PMID- 20018025 TI - Genome-wide analysis of haplotype interaction for the data from the North American Rheumatoid Arthritis Consortium. AB - Recent genome-wide association studies on several complex diseases have focused on individual single-nucleotide polymorphism (SNP) analysis; however, not many studies have reported interactions among genes perhaps because the gene-gene and gene-environment interaction analysis could be infeasible due to heavy computing requirements. In this study we propose a new strategy for exploring the interactions among haplotypes. The proposed method consists of two steps. Step 1 tests the single-SNP association of whole genome with multiple testing corrections and finds the haplotype blocks of the significant SNPs. Step 2 performs interaction analysis of haplotypes within blocks. Our proposed method is applied to the rheumatoid arthritis data for Genetic Analysis Workshop 16. PMID- 20018026 TI - Genome-wide association studies using single-nucleotide polymorphisms versus haplotypes: an empirical comparison with data from the North American Rheumatoid Arthritis Consortium. AB - The high genomic density of the single-nucleotide polymorphism (SNP) sets that are typically surveyed in genome-wide association studies (GWAS) now allows the application of haplotype-based methods. Although the choice of haplotype-based vs. individual-SNP approaches is expected to affect the results of association studies, few empirical comparisons of method performance have been reported on the genome-wide scale in the same set of individuals. To measure the relative ability of the two strategies to detect associations, we used a large dataset from the North American Rheumatoid Arthritis Consortium to: 1) partition the genome into haplotype blocks, 2) associate haplotypes with disease, and 3) compare the results with individual-SNP association mapping. Although some associations were shared across methods, each approach uniquely identified several strong candidate regions. Our results suggest that the application of both haplotype-based and individual-SNP testing to GWAS should be adopted as a routine procedure. PMID- 20018027 TI - Conditional analysis of the major histocompatibility complex in rheumatoid arthritis. AB - We performed a whole-genome association study of rheumatoid arthritis susceptibility using Illumina 550k single-nucleotide polymorphism (SNP) genotypes of 868 cases and 1194 controls from the North American Rheumatoid Arthritis Consortium (NARAC). Structured association analysis with adjustment for potential population stratification yielded 200 SNPs with p < 1 x 10-8 for association with RA, all of which were on chromosome 6 in a 2.7-Mb region of the major histocompatibility complex (MHC). Given the extensive linkage equilibrium in the region and known risk of HLA-DRB1 alleles, we then applied conditional analyses to ascertain independent signals for RA susceptibility among these 200 candidate SNPs. Conditional analyses incorporating risk categories of the HLA-DRB1 "shared epitope" revealed three SNPs having independent associations with RA (conditional p < 0.001). This supports the presence of significant effects on RA susceptibility in the MHC in addition to the shared epitope. PMID- 20018028 TI - Armitage's trend test for genome-wide association analysis: one-sided or two sided? AB - The importance of considering confounding due to population stratification in genome-wide association analysis using case-control designs has been a source of debate. Armitage's trend test, together with some other methods developed from it, can correct for population stratification to some extent. However, there is a question whether the one-sided or the two-sided alternative hypothesis is appropriate, or to put it another way, whether examining both the one-sided and the two-sided alternative hypotheses can give more information. The dataset for Problem 1 of Genetic Analysis Workshop 16 provides us with a chance to address this question. Because it is a part of a combined sample from the North American Rheumatoid Arthritis Consortium (NARAC) and the Swedish Epidemiological Investigation of Rheumatoid Arthritis (EIRA), the results from the combined sample can be used as references. To test this aim, the last 10,000 single nucleotide polymorphisms (SNPs) on chromosome 9, which contain the common genetic variant at the TRAF1-C5 locus, were examined by conducting Armitage's trend tests. Examining the two-sided alternative hypothesis shows that SNPs rs12380341 (p = 9.7 x 10-11) and rs872863 (p = 1.7 x 10-15), along with six SNPs across the TRAF1-C5 locus, rs1953126, rs10985073, rs881375, rs3761847, rs10760130, and rs2900180 (p~1 x 10-7), are significantly associated with anti-cyclic citrullinated peptide-positive rheumatoid arthritis. But examining the one-sided alternative hypothesis that the minor allele is positively associated with the disease shows that only those six SNPs across the TRAF1-C5 locus are significantly associated with the disease (p~1 x 10-8), which is consistent with the results from the combined sample of the NARAC and the EIRA. PMID- 20018029 TI - Genome-wide association studies of rheumatoid arthritis data via multiple hypothesis testing methods for correlated tests. AB - Genome-wide association studies often involve testing hundreds of thousands of single-nucleotide polymorphisms (SNPs). These tests may be highly correlated because of linkage disequilibrium among SNPs. Multiple testing correction ignoring the correlation among markers, as is done in the Bonferroni procedure, can cause loss of power. Several multiple testing adjustment methods accounting for correlations among tests have been developed and have shown improved power compared to the Bonferroni procedure. These methods include a Monte Carlo (MC) method and a method of computing p-values adjusted for correlated tests. The objective of this study is to apply these two multiple testing methods to genome wide association study of the Genetic Analysis Workshop 16 rheumatoid arthritis data from the North American Rheumatoid Arthritis Consortium, to compare the performance of these two methods to the Bonferroni procedure in identifying susceptibility loci underlying rheumatoid arthritis, and to discuss the strengths and weaknesses of these methods. The results show that both the MC method and p values adjusted for correlated tests method identified more significant SNPs, thus potentially have higher power than the corresponding Bonferroni methods using the same test statistics as in the MC method and p-values adjusted for correlated tests, respectively. Simulation studies demonstrate that the MC method may have slightly higher power than the p-values adjusted for correlated tests method. PMID- 20018030 TI - Non-redundant summary scores applied to the North American Rheumatoid Arthritis Consortium dataset. AB - After performing a genome-wide association study, it is often difficult to know which regions to follow up, especially when no one marker reaches genome-wide significance. Researchers frequently focus on their top N findings, knowing that true associations may be buried deeper in the list. Others focus on genes or regions that have multiple markers showing evidence of association. However, these markers are often in high linkage disequilibrium with one another (r2 > 0.80), which indicates that these additional markers are providing redundant information. I propose a novel method that identifies regions with multiple lines of evidence, by down-weighting the contribution of additional markers in proportion to pairwise linkage disequilibrium. I have used this non-redundant summary score in my analysis of the North American Rheumatoid Arthritis Consortium dataset released as part of Genetic Analysis Workshop 16. Three regions were identified that had a genome-wide empirical p-value less than 0.01, including one novel region on chromosome 20 near the KCNB1 and PTGIS genes. PMID- 20018031 TI - The Genetic Analysis Workshop 16 Problem 3: simulation of heritable longitudinal cardiovascular phenotypes based on actual genome-wide single-nucleotide polymorphisms in the Framingham Heart Study. AB - The Genetic Analysis Workshop (GAW) 16 Problem 3 comprises simulated phenotypes emulating the lipid domain and its contribution to cardiovascular disease risk. For each replication there were 6,476 subjects in families from the Framingham Heart Study (FHS), with their actual genotypes for Affymetrix 550 k single nucleotide polymorphisms (SNPs) and simulated phenotypes. Phenotypes are simulated at three visits, 10 years apart. There are up to 6 "major" genes influencing variation in high- and low-density lipoprotein cholesterol (HDL, LDL), and triglycerides (TG), and 1,000 "polygenes" simulated for each trait. Some polygenes have pleiotropic effects. The locus-specific heritabilities of the major genes range from 0.1 to 1.0%, under additive, dominant, or overdominant modes of inheritance. The locus-specific effects of the polygenes ranged from 0.002 to 0.15%, with effect sizes selected from negative exponential distributions. All polygenes act independently and have additive effects. Individuals in the LDL upper tail were designated medicated. Subjects medicated increased across visits at 2%, 5%, and 15%. Coronary artery calcification (CAC) was simulated using age, lipid levels, and CAC-specific polymorphisms. The risk of myocardial infarction before each visit was determined by CAC and its interactions with smoking and two genetic loci. Smoking was simulated to be commensurate with rates reported by the Centers for Disease Control. Two hundred replications were simulated. PMID- 20018032 TI - Using a higher criticism statistic to detect modest effects in a genome-wide study of rheumatoid arthritis. AB - In high-dimensional studies such as genome-wide association studies, the correction for multiple testing in order to control total type I error results in decreased power to detect modest effects. We present a new analytical approach based on the higher criticism statistic that allows identification of the presence of modest effects. We apply our method to the genome-wide study of rheumatoid arthritis provided in the Genetic Analysis Workshop 16 Problem 1 data set. There is evidence for unknown bias in this study that could be explained by the presence of undetected modest effects. We compared the asymptotic and empirical thresholds for the higher criticism statistic. Using the asymptotic threshold we detected the presence of modest effects genome-wide. We also detected modest effects using 90th percentile of the empirical null distribution as a threshold; however, there is no such evidence when the 95th and 99th percentiles were used. While the higher criticism method suggests that there is some evidence for modest effects, interpreting individual single-nucleotide polymorphisms with significant higher criticism statistics is of undermined value. The goal of higher criticism is to alert the researcher that genetic effects remain to be discovered and to promote the use of more targeted and powerful studies to detect the remaining effects. PMID- 20018033 TI - The effect of minor allele frequency on the likelihood of obtaining false positives. AB - Determining the most promising single-nucleotide polymorphisms (SNPs) presents a challenge in genome-wide association studies, when hundreds of thousands of association tests are conducted. The power to detect genetic effects is dependent on minor allele frequency (MAF), and genome-wide association studies SNP arrays include SNPs with a wide distribution of MAFs. Therefore, it is critical to understand MAF's effect on the false positive rate.Data from the Framingham Heart Study simulated data (Problem 3, with answers) was used to examine the effects of varying MAFs on the likelihood of false positives. Replication set 1 was used to generate 1 million permutations of case/control status in unrelated individuals. Logistic regression was used to test for the association between each SNP and myocardial infarction using an additive model. We report the number of "significant" tests by MAF at alpha = 10-4, 10-5, and 10-6.Common SNPs exhibited fewer false positives than expected. At alpha = 10-4, SNPs with MAF 25% and 50% resulted in 69.2 [95%CI: 62.8-75.6] and 70.8 [95%CI: 61.3-80.4] false positives, respectively, compared to 100 expected. Rare SNPs exhibited more variability but did not show more false-positive results than expected by chance. However, at alpha = 10-4, MAF = 5% exhibited significantly more false positives (105.5 [95%CI: 81-130.1]) than MAF = 25% and 50%. Similar results were seen at the other alpha values.These results suggest that removal of low MAF SNPs from analysis due to concerns about inflated false-positive results may not be appropriate. PMID- 20018034 TI - Multivariate association analysis of the components of metabolic syndrome from the Framingham Heart Study. AB - Metabolic syndrome, by definition, is the manifestation of multiple, correlated metabolic impairments. It is known to have both strong environmental and genetic contributions. However, isolating genetic variants predisposing to such a complex trait has limitations. Using pedigree data, when available, may well lead to increased ability to detect variants associated with such complex traits. The ability to incorporate multiple correlated traits into a joint analysis may also allow increased detection of associated genes. Therefore, to demonstrate the utility of both univariate and multivariate family-based association analysis and to identify possible genetic variants associated with metabolic syndrome, we performed a scan of the Affymetrix 50 k Human Gene Panel data using 1) each of the traits comprising metabolic syndrome: triglycerides, high-density lipoprotein, systolic blood pressure, diastolic blood pressure, blood glucose, and body mass index, and 2) a composite trait including all of the above, jointly. Two single-nucleotide polymorphisms within the cholesterol ester transfer protein (CETP) gene remained significant even after correcting for multiple testing in both the univariate (p < 5 x 10-7) and multivariate (p < 5 x 10-9) association analysis. Three genes met significance for multiple traits after correction for multiple testing in the univariate analysis, while five genes remained significant in the multivariate association. We conclude that while both univariate and multivariate family-based association analysis can identify genes of interest, our multivariate approach is less affected by multiple testing correction and yields more significant results. PMID- 20018035 TI - A combinatorial approach for detecting gene-gene interaction using multiple traits of Genetic Analysis Workshop 16 rheumatoid arthritis data. AB - Rheumatoid arthritis is inherited in a complex manner. So far several single susceptibility genes, such as PTPN22, STAT4, and TRAF1-C5, have been identified. However, it is presumed that some genes may interact to have a significant effect on the disease, while each of them only plays a modest role. We propose a new combinatorial association test to detect the gene-gene interaction in the rheumatoid arthritis data using multiple traits: disease status, anti-cyclic citrullinated peptide, and immunoglobulin M. Existing gene-gene interaction tests only use the information on a single trait at a time. In this article, we propose a new multivariate combinatorial searching method that utilizes multiple traits at the same time. Multivariate combinatorial searching method is conducted by incorporating the multiple traits with various techniques of feature selection to search for a set of disease-susceptibility genes that may interact. By analyzing three panels of markers, we have identified a significant gene-gene interaction between PTPN22 and TRAF1-C5. PMID- 20018036 TI - Using a latent growth curve model for an integrative assessment of the effects of genetic and environmental factors on multiple phenotypes. AB - We propose the use of latent growth curve model to assess the influence of genetic, environmental, demographic, and lifestyle factors on multiple phenotypes related to coronary heart disease. We model four quantitative traits (systolic blood pressure, high-density lipoprotein, low-density lipoprotein, and triglycerides) simultaneously in a multivariate framework that allows us to study their change over time, assess individual variation, and investigate cross phenotype relationships. Environmental, demographic, and lifestyle covariates are included at different levels of the model as time-varying or time-invariant, as appropriate. To investigate the change over time attributed to genetic factors, we use candidate markers that have previously been shown to be associated with the quantitative traits. We illustrate our approach using independent observations from the offspring cohort of the Framingham Heart Study data. PMID- 20018037 TI - Mendelian randomization in family data. AB - The phrase "mendelian randomization" has become associated with the use of genetic polymorphisms to uncover causal relationships between phenotypic variables. The statistical methods useful in mendelian randomization are known as instrumental variable techniques. We present an approach to instrumental variable estimation that is useful in family data and is robust to the use of weak instruments. We illustrate our method to measure the causal influence of low density lipoprotein on high-density lipoprotein, body mass index, triglycerides, and systolic blood pressure. We use the Framingham Heart Study data as distributed to participants in the Genetics Analysis Workshop 16. PMID- 20018038 TI - Evaluation of genetic risk scores for lipid levels using genome-wide markers in the Framingham Heart Study. AB - BACKGROUND: Multiple single-nucleotide polymorphisms have been associated with low-density lipoprotein cholesterol (LDL-C), high-density lipoprotein cholesterol (HDL-C), and triglyceride (TG) levels. In this paper, we evaluate a weighted and an unweighted approach for estimating the combined effect of multiple markers (using genotypes and haplotypes) on lipid levels for a given individual. METHODS: Using data from the Framingham Heart Study SHARe genome-wide association study, we tested genome-wide genotypes and haplotypes for association with lipid levels and constructed genetic risk scores (GRS) based on multiple markers that were weighted according to their estimated effects on LDL-C, HDL-C, and TG. These scores (GRS-LDL, GRS-HDL, and GRS-TG) were then evaluated for associations with LDL-C, HDL-C, and TG, and compared with results of an unweighted method based on risk-allele counts. For comparability of metrics, GRS variables were divided into quartiles. RESULTS: GRS-LDL quartiles were associated with LDL-C levels (p = 2.1 x 10-24), GRS-HDL quartiles with HDL-C (p = 5.9 x 10-22), and GRS-TG quartiles with TG (p = 5.4 x 10-25). In comparison, these p-values were considerably lower than those for the associations of the unweighted GRS quartiles for LDL-C (p = 3.6 x 10-7), HDL-C (p = 6.4 x 10-16), and TG (p = 4.1 x 10-10). CONCLUSION: GRS variables were highly predictive of LDL-C, HDL-C, and TG measurements, especially when weighted based on each marker's individual association with those intermediate risk phenotypes. The allele-count GRS approach that does not weight the GRS by individual marker associations was considerably less predictive of lipid and lipoprotein measures when the same genetic markers were utilized, suggesting that substantially more risk-associated genetic marker information is encapsulated by the weighted GRS variables. PMID- 20018039 TI - Associating multiple longitudinal traits with high-dimensional single-nucleotide polymorphism data: application to the Framingham Heart Study. AB - Cardiovascular diseases are associated with combinations of phenotypic traits, which are in turn caused by a combination of environmental and genetic factors. Because of the diversity of pathways that may lead to cardiovascular diseases, we examined the so-called intermediate phenotypes, which are often repeatedly measured. We developed a penalized nonlinear canonical correlation analysis to associate multiple repeatedly measured traits with high-dimensional single nucleotide polymorphism data. PMID- 20018040 TI - Genetic association studies and the effect of misclassification and selection bias in putative confounders. AB - Genetic epidemiology studies often adjust for numerous potential confounders, yet the influences of confounder misclassification and selection bias are rarely considered. We used simulated data to evaluate the effect of confounder misclassification and selection bias in a case-control study of incident myocardial infarction. We show that putative confounders traditionally included in genetic association studies do not alter effect estimates, even when excessive levels of misclassification are incorporated. Conversely, selection bias resulting from covariates affected by the single-nucleotide polymorphism of interest can bias effect estimates upward or downward. These results support careful consideration of how well a study population represents the target population because selection bias may result even when associations are modest. PMID- 20018041 TI - The effect of multiple genetic variants in predicting the risk of type 2 diabetes. AB - While recently performed genome-wide association studies have advanced the identification of genetic variants predisposing to type 2 diabetes (T2D), the potential application of these novel findings for disease prediction and prevention has not been well studied. Diabetes prediction and prevention have become urgent issues owing to the rapidly increasing prevalence of diabetes and its associated mortality, morbidity, and health care cost. New prediction approaches using genetic markers could facilitate early identification of high risk sub-groups of the population so that appropriate prevention methods could be effectively applied to delay, or even prevent, disease onset.This paper assessed 18 recently identified T2D loci for their potential role in diabetes prediction. We built a new predictive genetic test for T2D using the Framingham Heart Study dataset. Using logistic regression and 15 additional loci, the new test was slightly improved over the existing test using just three loci. A formal comparison between the two tests suggests no significant improvement. We further formed a predictive genetic test for identifying early onset T2D and found higher classification accuracy for this test, not only indicating that these 18 loci have great potential for predicting early onset T2D, but also suggesting that they may play important roles in causing early-onset T2D.To further improve the test's accuracy, we applied a newly developed nonparametric method capable of capturing high order interactions to the data, but it did not outperform a logistic regression that only considers single-locus effects. This could be explained by the absence of gene-gene interactions among the 18 loci. PMID- 20018043 TI - Defining genetic determinants of the Metabolic Syndrome in the Framingham Heart Study using association and structural equation modeling methods. AB - The Metabolic Syndrome (MetSyn), which is a clustering of traits including insulin resistance, obesity, hypertension and dyslipidemia, is estimated to have a substantial genetic component, yet few specific genetic targets have been identified. Factor analysis, a sub-type of structural equation modeling (SEM), has been used to model the complex relationships in MetSyn. Therefore, we aimed to define the genetic determinants of MetSyn in the Framingham Heart Study (Offspring Cohort, Exam 7) using the Affymetrix 50 k Human Gene Panel and three different approaches: 1) an association-based "one-SNP-at-a-time" analysis with MetSyn as a binary trait using the World Health Organization criteria; 2) an association-based "one-SNP-at-a-time" analysis with MetSyn as a continuous trait using second-order factor scores derived from four first-order factors; and, 3) a multivariate SEM analysis with MetSyn as a continuous, second-order factor modeled with multiple putative genes, which were represented by latent constructs defined using multiple SNPs in each gene. Results were similar between approaches in that CSMD1 SNPs were associated with MetSyn in Approaches 1 and 2; however, the effects of CSMD1 diminished in Approach 3 when modeled simultaneously with six other genes, most notably CETP and STARD13, which were strongly associated with the Lipids and MetSyn factors, respectively. We conclude that modeling multiple genes as latent constructs on first-order trait factors, most proximal to the gene's function with limited paths directly from genes to the second-order MetSyn factor, using SEM is the most viable approach toward understanding overall gene variation effects in the presence of multiple putative SNPs. PMID- 20018042 TI - Assessment of genotype imputation methods. AB - Several methods have been proposed to impute genotypes at untyped markers using observed genotypes and genetic data from a reference panel. We used the Genetic Analysis Workshop 16 rheumatoid arthritis case-control dataset to compare the performance of four of these imputation methods: IMPUTE, MACH, PLINK, and fastPHASE. We compared the methods' imputation error rates and performance of association tests using the imputed data, in the context of imputing completely untyped markers as well as imputing missing genotypes to combine two datasets genotyped at different sets of markers. As expected, all methods performed better for single-nucleotide polymorphisms (SNPs) in high linkage disequilibrium with genotyped SNPs. However, MACH and IMPUTE generated lower imputation error rates than fastPHASE and PLINK. Association tests based on allele "dosage" from MACH and tests based on the posterior probabilities from IMPUTE provided results closest to those based on complete data. However, in both situations, none of the imputation-based tests provide the same level of evidence of association as the complete data at SNPs strongly associated with disease. PMID- 20018044 TI - Transmission-ratio distortion in the Framingham Heart Study. AB - Transmission-ratio distortion (TRD) is a phenomenon in which the segregation of alleles does not obey Mendel's laws. As a simple example, a recessive locus that results in fetal lethality will result in live-born individuals sharing more alleles at this locus than expected under Mendel's laws. This could result in apparent linkage of the phenotype of 'being alive' to such a chromosomal regions. Further, this could result in false-positive linkage when 'affected-only' parametric or non-parametric linkage analysis is performed. Similarly, loci demonstrating TRD may be detectable in family-based association tests as deviant transmission of alleles. Therefore, TRD could result in confounding of family based association studies of diseases. The Framingham Heart Study data available for Genetic Analysis Workshop 16 is a suitable dataset to determine whether there are loci in the genome that reveal TRD because of the large number of individuals from families, the high-resolution genotyping, and the population-based nature of the study. We have used both genome-wide linkage and family-based association methods to determine whether there are loci that demonstrate TRD in the Framingham Heart Study. Family-based association analysis identified thousands of loci with apparent TRD. However, the vast majority of these are likely the result of genotyping errors with application of strict quality control criteria to the genotype data, and automated inspection of the intensity plots, we identify a small number of loci that may show true TRD, including rs1000548 in intron 6 of S antigen (arrestin, SAG) on chromosome 2 (p = 7 x 10-10). PMID- 20018045 TI - Genome-wide association analysis of Framingham Heart Study data for the Genetics Analysis Workshop 16: effects due to medication use. AB - Problems associated with medication use and the consequent effects on genome wide association analyses were explored using the Genetic Analysis Workshop 16 Problem 3 data. Lipid phenotypes were simulated in the Framingham Heart Study using several measured variables including causal genes (based on a 500 k SNP panel), smoking, dietary intake, and medication usage. We report a sensitivity analysis of how medication use (which artificially alters lipid values) affects association inferences. Associations were performed for LDL-c under seven different correction schemes: 1) ignore medication use entirely (no correction) and adjust for covariates; 2) delete medicated subjects then adjust for covariates; 3) include medication use (Yes/No) as a covariate during covariate adjustments; 4) correct raw values using clinical trials information then adjust for covariates; 5) correct raw values using the actual simulation protocol ("truth") then adjust for covariates; and 6-7) over-corrections (add arbitrary values) then adjust for covariates. Results indicate that failure to properly correct for medication usage can profoundly affect the heritability, and hence the association results. The empirical results yielded one genome-wide significant locus on chromosome 22 (RS2294207), consistent with the simulation protocol. This signal was detected under all schemes that corrected the raw values (clinical trials, simulation protocol, or over corrections), but was not detected under the first three adjustment schemes (ignore medication use, delete medicated individuals, use medication status as covariate). In summary, we confirm that failure to properly account for medication usage can have a profound impact on genetic associations. PMID- 20018046 TI - Genome-wide association study for empirically derived metabolic phenotypes in the Framingham Heart Study offspring cohort. AB - We used data reduction and clustering methods to identify five phenotypically homogeneous groups of study participants with similar profiles for cardiovascular disease risk factors. We constructed both qualitative (binary subgroup membership) and quantitative traits (probability of subgroup membership) for each individual. The Cluster 1 comprised individuals who were generally healthy and had some history of smoking. Cluster 2 was dropped from the analyses due to the preponderance of missing data. Cluster 3 was used as the control group, healthy non-smokers. Members of Cluster 4 had features of the metabolic syndrome and were generally not as obese as Cluster 5. Obesity was the hallmark of Cluster 5, the members of which also had some features of the metabolic syndrome.We then examined the genetic associations with both qualitative and quantitative representations of these empirically derived traits. Genetic analyses of the qualitative traits were conducted, comparing each of the affected groups with the unaffected cluster alone and, to increase statistical power, the unaffected group and healthy smokers combined. One single-nucleotide polymorphism on chromosome 4 met a conservative genome-wide significance level, but the effect was muted when we accounted for population stratification. The results for the quantitative traits were similar, with a small number of genome-wide significant findings muted by control for admixture. The directional findings will provide the basis for hypothesis generation for syndromes such as the metabolic syndrome and obesity. PMID- 20018047 TI - Memory management in genome-wide association studies. AB - Genome-wide association is a powerful tool for the identification of genes that underlie common diseases. Genome-wide association studies generate billions of genotypes and pose significant computational challenges for most users including limited computer memory. We applied a recently developed memory management tool to two analyses of North American Rheumatoid Arthritis Consortium studies and measured the performance in terms of central processing unit and memory usage. We conclude that our memory management approach is simple, efficient, and effective for genome-wide association studies. PMID- 20018048 TI - Inference of disease associations with unmeasured genetic variants by combining results from genome-wide association studies with linkage disequilibrium patterns in a reference data set. AB - Results from whole-genome association studies of many common diseases are now available. Increasingly, these are being incorporated into meta-analyses to increase the power to detect weak associations with measured single-nucleotide polymorphisms (SNPs). Imputation of genotypes at unmeasured loci has been widely applied using patterns of linkage disequilibrium (LD) observed in the HapMap panels, but there is a need for alternative methods that can utilize the pooled effect estimates from meta-analyses and explore possible associations with SNPs and haplotypes that are not included in HapMap.By a weighted average technique, we show that association results for common SNPs in an observed data set can be scaled and combined to infer the effect of a genetic variant that has been measured only in an independent reference data set. We show that the ratio p(R 1)/[1 + p(R-1)], where R is the relative risk associated with a measured or unmeasured allele of frequency p, is appropriately scaled by 1/D' and weighted in proportion to r2, both common measures of LD being derived from the reference data set.We illustrate this computationally simple method by combining the results of a genome-wide association screen from the North American Rheumatoid Arthritis Consortium with LD measures from the British 1958 Birth Cohort, and explore the validity of underlying assumptions about the generalizability of LD from one population to another, and from healthy subjects to subjects with clinical disease. PMID- 20018049 TI - Evaluation of an optimal receiver operating characteristic procedure. AB - : Lu and Elston have recently proposed a procedure for developing optimal receiver operating characteristic curves that maximize the area under a receiver operating characteristic curve in the setting of a predictive genetic test. The method requires only summary data, not individual level genetic data. In an era of increased data sharing, we investigate the performance of this algorithm when individual level genetic data are available and compare this approach to more standard receiver operating characteristic curve-building methods. CONCLUSION: Though the Lu-Elston method can produce an optimal area under the curve under some assumptions, the method typically has little advantage over standard multivariable logistic methods when data are available. Also, the standard approach easily allows comparison of nested models via likelihood ratio tests and incorporation of covariates - the Lu-Elston approach is shown to have some difficulties with such analyses. These conclusions are based on evaluations using the Genetic Analysis Workshop 16 rheumatoid arthritis data set. PMID- 20018050 TI - Application of sex-specific single-nucleotide polymorphism filters in genome-wide association data. AB - We explored five sex-specific quality control filters in North American Rheumatoid Arthritis Consortium's Illumina 550 k datasets. Three X chromosome and three autosomal single-nucleotide polymorphisms flagged by sex quality control filters were missed by filters of call rate at 95% and Hardy-Weinberg equilibrium at 10-6. We applied a subset of these sex-specific quality control filters to eight chromosomes in the Framingham Heart Study samples genotyped by Affymetrix 500 k SNP arrays, and identified another two single-nucleotide polymorphisms that failed to be picked up by the above global filters. PMID- 20018051 TI - ACPA: automated cluster plot analysis of genotype data. AB - Genome-wide association studies have become standard in genetic epidemiology. Analyzing hundreds of thousands of markers simultaneously imposes some challenges for statisticians. One issue is the problem of multiplicity, which has been compared with the search for the needle in a haystack. To reduce the number of false-positive findings, a number of quality filters such as exclusion of single nucleotide polymorphisms (SNPs) with a high missing fraction are employed. Another filter is exclusion of SNPs for which the calling algorithm had difficulties in assigning the genotypes. The only way to do this is the visual inspection of the cluster plots, also termed signal intensity plots, but this approach is often neglected. We developed an algorithm ACPA (automated cluster plot analysis), which performs this task automatically for autosomal SNPs. It is based on counting samples that lie too close to the cluster of a different genotype; SNPs are excluded when a certain threshold is exceeded. We evaluated ACPA using 1,000 randomly selected quality controlled SNPs from the Framingham Heart Study data that were provided for the Genetic Analysis Workshop 16. We compared the decision of ACPA with the decision made by two independent readers. We achieved a sensitivity of 88% (95% CI: 81%-93%) and a specificity of 86% (95% CI: 83%-89%). In a screening setting in which one aims at not losing any good SNP, we achieved 99% (95% CI: 98%-100%) specificity and still detected every second low-quality SNP. PMID- 20018052 TI - Look who is calling: a comparison of genotype calling algorithms. AB - In genome-wide association studies, high-level statistical analyses rely on the validity of the called genotypes, and different genotype calling algorithms (GCAs) have been proposed. We compared the GCAs Bayesian robust linear modeling using Mahalanobis distance (BRLMM), Chiamo++, and JAPL using the autosomal single nucleotide polymorphisms (SNPs) from the 500 k Affymetrix Array Set data of the Framingham Heart Study as provided for the Genetic Analysis Workshop 16, Problem 2, and prepared standard quality control (sQC) for each algorithm. Using JAPL, most individuals were retained for the analysis. The lowest number of SNPs that successfully passed sQC was observed for BRLMM and the highest for Chiamo++. All three GCAs fulfilled all sQC criteria for 79% of the SNPs but at least one GCA failed for 18% of the SNPs. Previously undetected errors in strand coding were identified by comparing genotype concordances between GCAs. Concordance dropped with the number of GCAs failing sQC. We conclude that JAPL and Chiamo++ are the GCAs of choice if the aim is to keep as many subjects and SNPs as possible, respectively. PMID- 20018053 TI - A genome-wide association scan for rheumatoid arthritis data by Hotelling's T2 tests. AB - We performed a genome-wide association scan on the North American Rheumatoid Arthritis Consortium (NARAC) data using Hotelling's T2 tests, i.e., TH based on allele coding and TG based on genotype coding. The objective was to identify associations between single-nucleotide polymorphisms (SNPs) or markers and rheumatoid arthritis. In specific candidate gene regions, we evaluated the performance of Hotelling's T2 tests. Then Hotelling's T2 tests were used as a tool to identify new regions that contain SNPs showing strong associations with disease. As expected, the strongest association evidence was found in the region of the HLA-DRB1 locus on chromosome 6. In the region of the TRAF1-C5 genes, we identified two SNPs, rs2900180 and rs3761847, with the largest and the second largest TH and TG scores among all SNPs on chromosome 9. We also identified one SNP, rs2476601, in the region of the PTPN22 gene that had the largest TH score and the second largest TG score among all SNPs on chromosome 1. In addition, SNPs with the largest TH score on each chromosome were identified. These SNPs may be located in the regions of genes that have modest effects on rheumatoid arthritis. These regions deserve further investigation. PMID- 20018054 TI - Predictive modeling in case-control single-nucleotide polymorphism studies in the presence of population stratification: a case study using Genetic Analysis Workshop 16 Problem 1 dataset. AB - In this paper, we apply the gradient-boosting machine predictive model to the rheumatoid arthritis data for predicting the case-control status. QQ-plot suggests severe population stratification. In univariate genome-wide association studies, a correction factor for ethnicity confounding can be derived. Here we propose a novel strategy to deal with population stratification in the context of multivariate predictive modeling. We address the problem by clustering the subjects on the axes of genetic variations, and building a predictive model separately in each cluster. This allows us to control ethnicity without explicitly including it in the model, which could marginalize the genetic signal we are trying to discover. Clustering not only leads to more similar ethnicity groups but also, as our results show, increases the accuracy of our model when compared to the non-clustered approach. The highest accuracy is achieved with the model adjusted for population stratification, when the genetic axes of variation are included among the set of predictors, although this may be misleading given the confounding effects. PMID- 20018055 TI - Analysis of North American Rheumatoid Arthritis Consortium data using a penalized logistic regression approach. AB - We applied a penalized regression approach to single-nucleotide polymorphisms in regions on chromosomes 1, 6, and 9 of the North American Rheumatoid Arthritis Consortium data. Results were compared with a standard single-locus association test. Overall, the penalized regression approach did not appear to offer any advantage with respect to either detection or localization of disease-associated polymorphisms, compared with the single-locus approach. PMID- 20018056 TI - Combining least absolute shrinkage and selection operator (LASSO) and principal components analysis for detection of gene-gene interactions in genome-wide association studies. AB - Variable selection in genome-wide association studies can be a daunting task and statistically challenging because there are more variables than subjects. We propose an approach that uses principal-component analysis (PCA) and least absolute shrinkage and selection operator (LASSO) to identify gene-gene interaction in genome-wide association studies. A PCA was used to first reduce the dimension of the single-nucleotide polymorphisms (SNPs) within each gene. The interaction of the gene PCA scores were placed into LASSO to determine whether any gene-gene signals exist. We have extended the PCA-LASSO approach using the bootstrap to estimate the standard errors and confidence intervals of the LASSO coefficient estimates. This method was compared to placing the raw SNP values into the LASSO and the logistic model with individual gene-gene interaction. We demonstrated these methods with the Genetic Analysis Workshop 16 rheumatoid arthritis genome-wide association study data and our results identified a few gene-gene signals. Based on our results, the PCA-LASSO method shows promise in identifying gene-gene interactions, and, at this time we suggest using it with other conventional approaches, such as generalized linear models, to narrow down genetic signals. PMID- 20018057 TI - Detecting single-nucleotide polymorphism by single-nucleotide polymorphism interactions in rheumatoid arthritis using a two-step approach with machine learning and a Bayesian threshold least absolute shrinkage and selection operator (LASSO) model. AB - The objective of this study was to detect interactions between relevant single nucleotide polymorphisms (SNPs) associated with rheumatoid arthritis (RA). Data from Problem 1 of the Genetic Analysis Workshop 16 were used. These data consisted of 868 cases and 1,194 controls genotyped with the 500 k Illumina chip. First, machine learning methods were applied for preselecting SNPs. One hundred SNPs outside the HLA region and 1,500 SNPs in the HLA region were preselected using information-gain theory. The software weka was used to reduce colinearity and redundancy in the HLA region, resulting in a subset of 6 SNPs out of 1,500. In a second step, a parametric approach to account for interactions between SNPs in the HLA region, as well as HLA-nonHLA interactions was conducted using a Bayesian threshold least absolute shrinkage and selection operator (LASSO) model incorporating 2,560 covariates. This approach detected some main and interaction effects for SNPs in genes that have previously been associated with RA (e.g., rs2395175, rs660895, rs10484560, and rs2476601). Further, some other SNPs detected in this study may be considered in candidate gene studies. PMID- 20018058 TI - Evaluation of random forests performance for genome-wide association studies in the presence of interaction effects. AB - Random forests (RF) is one of a broad class of machine learning methods that are able to deal with large-scale data without model specification, which makes it an attractive method for genome-wide association studies (GWAS). The performance of RF and other association methods in the presence of interactions was evaluated using the simulated data from Genetic Analysis Workshop 16 Problem 3, with knowledge of the major causative markers, risk factors, and their interactions in the simulated traits. There was good power to detect the environmental risk factors using RF, trend tests, or regression analyses but the power to detect the effects of the causal markers was poor for all methods. The causal marker that had an interactive effect with smoking did show moderate evidence of association in the RF and regression analyses, suggesting that RF may perform well at detecting such interactions in larger, more highly powered datasets. PMID- 20018060 TI - The difficulties of reproducing conventionally derived results through 500k-chip technology. AB - Based on a "training" sample of 1,042 subjects genotyped for 5,728 single nucleotide polymorphisms (SNPs) of a conventional 0.4-Mb genome scan and a "test" sample of 746 subjects genotyped for 545,080 SNPs on a 500k-chip, we investigated the extent to which the subjects' immunoglobulin M levels can be reproducibly predicted from a multilocus genotype. We were specifically interested in the reproducibility of predictors across populations (1,042 versus 746 subjects) and across SNP sets (conventional genome scan versus anonymous 500k-chip) because this is a prerequisite for clinical application. For the training sample, neural network (NN) analysis yielded classifiers that predicted immunoglobulin M levels from the subjects' multilocus genotypes at acceptable error rates through a configuration of 15 genomic loci (61 SNPs). With the test sample (746 subjects) we addressed the question of reproducibility across populations and across SNP sets by means of a novel "competitive SNP set" approach. However, the chip data contained several sources of distortion, including greatly elevated noise levels and artifact-prone SNP regions, thus complicating attempts to verify the reproducibility of NN predictors. Though 5 of 15 genomic loci from the training samples appeared to be reproducible, the NN classifiers derived so far from the test samples are insufficiently compatible with the training samples. Nonetheless, our results are promising enough to justify further investigations. Because the underlying algorithm can easily be split into parallel tasks, the proposed "competitive SNP set" approach has turned out to be well suited for computers with today's 64-bit multiprocessor architectures and to offer a valuable extension to genome-wide association analyses. PMID- 20018059 TI - Evaluation of single-nucleotide polymorphism imputation using random forests. AB - Genome-wide association studies (GWAS) have helped to reveal genetic mechanisms of complex diseases. Although commonly used genotyping technology enables us to determine up to a million single-nucleotide polymorphisms (SNPs), causative variants are typically not genotyped directly. A favored approach to increase the power of genome-wide association studies is to impute the untyped SNPs using more complete genotype data of a reference population.Random forests (RF) provides an internal method for replacing missing genotypes. A forest of classification trees is used to determine similarities of probands regarding their genotypes. These proximities are then used to impute genotypes of untyped SNPs.We evaluated this approach using genotype data of the Framingham Heart Study provided as Problem 2 for Genetic Analysis Workshop 16 and the Caucasian HapMap samples as reference population. Our results indicate that RFs are faster but less accurate than alternative approaches for imputing untyped SNPs. PMID- 20018061 TI - Identification of correlated genetic variants jointly associated with rheumatoid arthritis using ridge regression. AB - Using the North American Rheumatoid Arthritis Consortium genome-wide association dataset, we applied ridged, multiple least-squares regression to identify genetic variants with apparent unique contributions to variation of anti-cyclic citrullinated peptide (anti-CCP), a newly identified clinical risk factor for development of rheumatoid arthritis. Within a 2.7-Mbp region on chromosome 6 around the well studied HLA-DRB1 locus, ridge regression identified a single nucleotide polymorphism that was associated with anti-CCP variation when including the additive effects of other single-nucleotide polymorphisms in a multivariable analysis, but that showed only a weak direct association with anti CCP. This suggests that multivariable methods can be used to identify potentially relevant genetic variants in regions of interest that would be difficult to detect based on direct associations. PMID- 20018062 TI - Identification of genes and haplotypes that predict rheumatoid arthritis using random forests. AB - Random forest (RF) analysis of genetic data does not require specification of the mode of inheritance, and provides measures of variable importance that incorporate interaction effects. In this paper we describe RF-based approaches for assessment of gene and haplotype importance, and apply these approaches to a subset of the North American Rheumatoid Arthritis Consortium case-control data provided by Genetic Analysis Workshop 16. The RF analyses of 37 genes identified many of the same genes as logistic regression, but also suggested importance of certain single-nucleotide polymorphism and genes that were not ranked highly by logistic regression. A new permutation method did not reveal strong evidence of gene-gene interaction effects in these data. Although RFs are a promising approach for genetic data analysis, extensions beyond simple single-nucleotide polymorphism analyses and modifications to improve computational feasibility are needed. PMID- 20018063 TI - Detecting significant single-nucleotide polymorphisms in a rheumatoid arthritis study using random forests. AB - Random forest is an efficient approach for investigating not only the effects of individual markers on a trait but also the effect of the interactions among the markers in genetic association studies. This approach is especially appealing for the analysis of genome-wide data, such as those obtained from gene expression/single-nucleotide polymorphism (SNP) array experiments in which the number of candidate genes/SNPs is vast. We applied this approach to the Genetic Analysis Workshop 16 Problem 1 data to identify SNPs that contribute to rheumatoid arthritis. The random forest computed a raw importance score for each SNP marker, where higher importance score suggests higher level of association between the marker and the trait. The significance level of the association was determined empirically by repeatedly reapplying the random forest on randomly generated data under the null hypothesis that no association exists between the markers and the trait. Using random forest, we were able to identify 228 significant SNPs (at the genome-wide significant level of 0.05) across the whole genome, over two-thirds of which are located on chromosome 6, especially clustered in the region of 6p21 containing the human leukocyte antigen (HLA) genes, such as gene HLA-DRB1 and HLA-DRA. Further analysis of this region indicates a strong association to the rheumatoid arthritis status. PMID- 20018064 TI - Single versus multiple imputation for genotypic data. AB - Due to the growing need to combine data across multiple studies and to impute untyped markers based on a reference sample, several analytical tools for imputation and analysis of missing genotypes have been developed. Current imputation methods rely on single imputation, which ignores the variation in estimation due to imputation. An alternative to single imputation is multiple imputation. In this paper, we assess the variation in imputation by completing both single and multiple imputations of genotypic data using MACH, a commonly used hidden Markov model imputation method. Using data from the North American Rheumatoid Arthritis Consortium genome-wide study, the use of single and multiple imputation was assessed in four regions of chromosome 1 with varying levels of linkage disequilibrium and association signals. Two scenarios for missing genotypic data were assessed: imputation of untyped markers and combination of genotypic data from two studies. This limited study involving four regions indicates that, contrary to expectations, multiple imputations may not be necessary. PMID- 20018065 TI - Selection of important variables by statistical learning in genome-wide association analysis. AB - Genetic analysis of complex diseases demands novel analytical methods to interpret data collected on thousands of variables by genome-wide association studies. The complexity of such analysis is multiplied when one has to consider interaction effects, be they among the genetic variations (G x G) or with environment risk factors (G x E). Several statistical learning methods seem quite promising in this context. Herein we consider applications of two such methods, random forest and Bayesian networks, to the simulated dataset for Genetic Analysis Workshop 16 Problem 3. Our evaluation study showed that an iterative search based on the random forest approach has the potential in selecting important variables, while Bayesian networks can capture some of the underlying causal relationships. PMID- 20018066 TI - Epistatic interactions of CDKN2B-TCF7L2 for risk of type 2 diabetes and of CDKN2B JAZF1 for triglyceride/high-density lipoprotein ratio longitudinal change: evidence from the Framingham Heart Study. AB - Fifteen known type 2 diabetes (T2D) gene variants were assessed for their associations with T2D status in 228 T2D families from the Framingham Heart Study (FHS) Original, Offspring, and Children Cohorts. Bayesian approach was used to test single-single-nucleotide polymorphism (SNP) association followed by logistic regression. Bayesian and logic regression approaches were used to test multiple SNP association searching for the best combinations of variants followed by logistic regression reconfirmation. The significant variants for T2D risk were also tested for their main and interacting effects on triglyceride (TG)/high density lipoprotein (HDL) ratio change derived from four point measures across time. This slope phenotype was made available using mixed model growth curve approach from 155 T2D families in the FHS Offspring Cohort. RESULTS: CDKN2B rs10811661 (p = 0.042), TCF7L2 rs4506565 (p = 0.004), and JAZF1 rs864745 (p = 0.04) were individually associated with risk of T2D (OR = 1.0-2.0; effect size <1%). CDKN2B and TCF7L2 were found with significant main (p = 0.02, 0.01) and interacting (p = 0.05) effects for increased (OR = 3.0) risk of T2D. CDKN2B and JAZF1 were found with significant main (p = 0.0002 and 0.034) and interacting (p = 0.001) effects on increased (beta = 0.42) TG/HDL ratio longitudinal change. These interacting effects were independent of effects of age and sex with effect sizes of 0.3-0.4% for risk of T2D or TG/HDL ratio longitudinal change. CONCLUSION: These synthetic approaches allowed for successful detection of CDKN2B and TCF7L2 interacting effect for T2D risk and CDKN2B and JAZF1 interacting effect on TG/HDL ratio increase over time among T2D families in the FHS. These interacting effects were consistent in conferring risk of T2D or progressive insulin resistance with modest effect sizes. PMID- 20018067 TI - A two-stage search strategy for detecting multiple loci associated with rheumatoid arthritis. AB - Gene x gene interactions play important roles in the etiology of complex multi factorial diseases like rheumatoid arthritis (RA). In this paper, we describe our use of a two-stage search strategy consisting of information theoretic methods and logistic regression to detect gene x gene interactions associated with RA using the data in Problem 1 of Genetic Analysis Workshop 16. Our method detected interactions of several SNPs (single-SNP and SNP x SNP) that are located on chromosomal regions linked to RA and related diseases in previous studies. PMID- 20018068 TI - A comparison of case-only designs for detecting gene x gene interaction in rheumatoid arthritis using genome-wide case-control data in Genetic Analysis Workshop 16. AB - We compare and contrast case-only designs for detecting gene x gene (G x G) interaction in rheumatoid arthritis (RA) using the genome-wide data provided by Genetic Analysis Workshop 16 Problem 1. Logistic as well as novel multinomial and proportional odds models that do not depend on the specification of additive or dominant models for susceptibility loci were applied to the case-only sample. We identified 519 significant interactions (p < 1 x 10-4 in at least one test). All methods detected unique significant interactions; 169 were common to more than one model and only 21 were common to all models. Results emphasize that categorization of the genetic variables and choice of regression model are critical and hugely influential in the identification of G x G. Porportional odds and multinomial methods provide new tools for identification of G x G interactions. PMID- 20018069 TI - Power and false-positive rates for the restricted partition method (RPM) in a large candidate gene data set. AB - Many phenotypes of public health importance (e.g., diabetes, coronary artery disease, major depression, obesity, and addictions to alcohol and nicotine) involve complex pathways of action. Interactions between genetic variants or between genetic variants and environmental factors likely play important roles in the functioning of these pathways. Unfortunately, complex interacting systems are likely to have important interacting factors that may not readily reveal themselves to univariate analyses. Instead, detecting the role of some of these factors may require analyses that are sensitive to interaction effects.In this study, we evaluate the sensitivity and specificity of the restricted partition method (RPM) to detect signals related to coronary artery disease in the Genetic Analysis Workshop 16 Problem 3 data using the 50,000 k candidate gene single nucleotide polymorphism set. Power and false-positive rates were evaluated using the first 100 replicate datasets. This included an exploration of the utility of using of all genotyped family members compared with selecting one member per family. PMID- 20018070 TI - Rheumatoid arthritis-associated gene-gene interaction network for rheumatoid arthritis candidate genes. AB - Rheumatoid arthritis (RA, MIM 180300) is a chronic and complex autoimmune disease. Using the North American Rheumatoid Arthritis Consortium (NARAC) data set provided in Genetic Analysis Workshop 16 (GAW16), we used the genotype-trait distortion (GTD) scores and proposed analysis procedures to capture the gene-gene interaction effects of multiple susceptibility gene regions on RA. In this paper, we focused on 27 RA candidate gene regions (531 SNPs) based on a literature search. Statistical significance was evaluated using 1000 permutations. HLADRB1 was found to have strong marginal association with RA. We identified 14 significant interactions (p < 0.01), which were aggregated into an association network among 12 selected candidate genes PADI4, FCGR3, TNFRSF1B, ITGAV, BTLA, SLC22A4, IL3, VEGF, TNF, NFKBIL1, TRAF1-C5, and MIF. Based on our and other contributors' findings during the GAW16 conference, we further studied 24 candidate regions with 336 SNPs. We found 23 significant interactions (p-value < 0.01), nine interactions in addition to our initial findings, and the association network was extended to include candidate genes HLA-A, HLA-B, HLA-C, CTLA4, and IL6. As we will discuss in this paper, the reported possible interactions between genes may suggest potential biological activities of RA. PMID- 20018071 TI - Allelic based gene-gene interactions in rheumatoid arthritis. AB - The detection of gene-gene interaction is an important approach to understand the etiology of rheumatoid arthritis (RA). The goal of this study is to identify gene-gene interaction of SNPs at the allelic level contributing to RA using real data sets (Problem 1) of North American Rheumatoid Arthritis Consortium (NARAC) provided by Genetic Analysis Workshop 16 (GAW16). We applied our novel method that can detect the interaction by a definition of nonrandom association of alleles that occurs when the contribution to RA of a particular allele inherited in one gene depends on a particular allele inherited at other unlinked genes. Starting with 639 single-nucleotide polymorphisms (SNPs) from 26 candidate genes, we identified ten two-way interacting genes and one case of three-way interacting genes. SNP rs2476601 on PTPN22 interacts with rs2306772 on SLC22A4, which interacts with rs881372 on TRAF1 and rs2900180 on C5, respectively. SNP rs2900180 on C5 interacts with rs2242720 on RUNX1, which interacts with rs881375 on TRAF1. Furthermore, rs2476601 on PTPN22 also interacts with three SNPs (rs2905325, rs1476482, and rs2106549) in linkage disequilibrium (LD) on IL6. The other three SNPs (rs2961280, rs2961283, and rs2905308) in LD on IL6 interact with two SNPs (rs477515 and rs2516049) on HLA-DRB1. SNPs rs660895 and rs532098 on HLA-DRB1 interact with rs2834779 and four SNPs in LD on RUNX1. Three-way interacting genes of rs10229203 on IL6, rs4816502 on RUNX1, and rs10818500 on C5 were also detected. PMID- 20018072 TI - Different models and single-nucleotide polymorphisms signal the simulated weak gene-gene interaction for a quantitative trait using haplotype-based and mixed models testing. AB - Knowledge of simulated genetic effects facilitates interpretation of methodological studies. Genetic interactions for common disorders are likely numerous and weak. Using the 200 replicates of the Genetic Analysis Workshop 16 (GAW16) Problem 3 simulated data, we compared the statistical power to detect weak gene-gene interactions using a haplotype-based test in the UNPHASED software with genotypic mixed model (GMM) and additive mixed model (AMM) mixed linear regression model in SAS. We assumed a candidate-gene approach where a single nucleotide polymorphism (SNP) in one gene is fixed and multiple SNPs are at the second gene. We analyzed the quantitative low-density lipoprotein trait (heritability 0.7%), modulated by simulated interaction of rs4648068 from 4q24 and another gene on 8p22, where we analyzed seven SNPs. We generally observed low power calculated per SNP ( or =45 years), the birth month effect was put to the test, for both lifetime fertility as well as child survival, controlling for maternal birth cohort (1826-1835, 1836 1845, 1846-1855), Duncan's SEI, urbanity, nativity, literacy and marital duration. Testing for potential cohort effects did not indicate a temporal trend in fertility by maternal birth month (seasonal Mann-Kendall test, p=0.578), while a minute increase in offspring survival was detected (p<0.001, Sen's estimator of slope=0.02, 95% CI=0.02 to 0.03). Further analyses of the maternal birth month effect on child survival were therefore seized. For lifetime fertility, ANOVA results indicated that maternal birth month was a major predictor for total offspring count (F11, 33606=1809.0, p<0.001), accounting for 37.2% of the total variability. In addition to main effects, a statistically significant interaction effect was observed (F538, 33606=2.2, p<0.001), with a corresponding effect size of eta2=0.40. Planned contrasts revealed that birth-month-specific differences in fertility achieved statistical significance (F11, 31798=1712.9, p<0.001), while post-hoc multiple comparisons for literacy and nativity displayed an inverse relationship with fertility, which meets demographic expectations. Controlling for all factors of interest, models of cohort-specific offspring counts (independent ANOVAs for 1826-1835: F157, 3467=26.3, p<0.001; 1836-1845: F182, 10299=75.5, p<0.001; 1846-1855: F199, 19859=137.9, p<0.001) indicated that women born in the first half of the year (particularly, January, February, April and May) achieved above-average parity, while those born in the latter half (namely, July, October, November and December) displayed markedly lower fertility averages. These monthly disparities are in line with previous observations and appear to be linked to seasonal optimal ripening of the oocyte or seasonal preovulatory over-ripeness ovopathy (Jongbloet, 1992). PMID- 20018120 TI - Greenlandic schoolchildren's compliance with national dietary guidelines. AB - OBJECTIVE: The aim of the present study was to examine to what extent children and adolescents in Greenland comply with the national dietary guidelines, and to analyse the influence of habitation and family affluence on the compliance with dietary guidelines. DESIGN: Data were from the Health Behaviour in School-aged Children (HBSC) survey in Greenland. The 2006 survey included 2462 students aged 11 to 17 years. RESULTS: The proportion of students complying with the national dietary guidelines varied from 14 % to 87 % depending on the food item. Sweets and soft drinks had the lowest compliance. The oldest children had the following characteristics compared with the younger children: fewer ate traditional Greenlandic foods, fewer ate fruit, fewer ate breakfast daily on school days and more drank soft drinks frequently. More boys than girls ate traditional Greenlandic foods often, while more girls ate vegetables daily. The least favourable eating habits in general were found among children from low affluent families and children in villages. CONCLUSIONS: Many Greenlandic schoolchildren did not comply with the national dietary guidelines. Despite a higher intake of traditional foods as a whole, children in villages and less affluent children were less likely to comply with guidelines. A strong relationship between diet, family affluence and availability was found. The study findings indicate that factors such as availability, cost and seasonal variation are important to the intake of both imported and traditional Greenlandic foods. The findings should be taken into consideration when promoting the national guidelines. PMID- 20018121 TI - The relationship between physical activity and mental health varies across activity intensity levels and dimensions of mental health among women and men. AB - OBJECTIVE: To explore gender-specific variations related to activity intensity in the relationship between physical activity (PA) and mental health (MH). Evaluating whether psychological well-being enhances with increases in PA at recommended levels and above, in the general population. DESIGN: Cross-sectional. SETTING: Population-based, representative for Belgium. SUBJECTS: A total of 6803 adults aged 25-64 years from the Belgian National Health Interview Survey. RESULTS: Multiple logistic regression analyses showed that clearly different intensity levels characterised the PA that associated with MH in women and men. In men, inverse associations existed between participation in vigorous-intensity PA and feelings of depression (OR = 0.580; 95 % CI 0.405, 0.830), anxiety (OR = 0.547; 95 % CI 0.364, 0.821) and symptoms of somatisation (OR = 0.590; 95 % CI 0.398, 0.874). In women, positive associations existed between walking and emotional well-being (OR = 1.202; 95 % CI 1.038, 1.394) and inverse associations between participation in moderate-intensity PA and symptoms of somatisation (OR = 0.737; 95 % CI 0.556, 0.977). Secondary analyses confirmed that differences in psychological complaints were significant for vigorous PA in men, and for moderate PA in women, whereas differences in emotional well-being were significant for walking exclusively in women. CONCLUSIONS: In the general population, the PA-MH relationship is always positive, regardless of activity intensity. In men, it addresses complaints (symptoms, palpable discomfort) and the optimal PA intensity is high. In women, it addresses complaints, but also distress (lowered mood, disturbing anxiety, altered well-being) and the PA intensity is mild. PMID- 20018122 TI - Validation of an FFQ to estimate the intake of fatty acids using erythrocyte membrane fatty acids and multiple 3d dietary records. AB - OBJECTIVE: The estimation of dietary intake in population-based studies is often assessed by the FFQ. The objective of our study is to evaluate the validity of an FFQ used to assess dietary fatty acid intake among middle-aged Chinese adults in Southern China. DESIGN: The method of triads was applied to obtain the validity coefficients (VC) of the FFQ for specific fatty acids. A subsample was randomly selected from an earlier cross-sectional study. The FFQ and 3d dietary records were used for dietary assessment, and the fatty acid composition of erythrocyte membranes was determined as the biomarker. RESULTS: The Spearman correlation coefficients between the FFQ and 3d dietary records were moderate to good (r = 0.28-0.66). The VC of the FFQ estimated by the method of triads were 0.72, 0.61, 0.65, 0.75 and 0.67 for MUFA, total n-6 fatty acids, alpha-linolenic acid, EPA and DHA, respectively. The VC could not be calculated for SFA, PUFA and total n-3 fatty acids because of negative correlations among the three measurements. But, the correlations between the FFQ and the dietary records were moderate for these fatty acids. CONCLUSIONS: Our FFQ applied in Southern Chinese adults was valid to estimate their dietary fatty acid intake and was thus suitable for use in a large cohort study. PMID- 20018123 TI - Television food advertising and the prevalence of childhood overweight and obesity: a multicountry comparison. AB - OBJECTIVE: To estimate the contribution of television (TV) food advertising to the prevalence of obesity among 6-11-year-old children in Australia, Great Britain (England and Scotland only), Italy, The Netherlands, Sweden and the United States. DESIGN: Data from contemporary representative studies on the prevalence of childhood obesity and on TV food advertising exposure in the above countries were entered into a mathematical simulation model. Two different effect estimators were used to calculate the reduction in prevalence of overweight and obesity in the absence of TV food advertising in each country; one based on literature and one based on experts' estimates. SETTING: Six- to eleven-year-old children in six Western countries. RESULTS: Estimates of the average exposure of children to TV food advertising range from 1.8 min/d in The Netherlands to 11.5 min/d in the United States. Its contribution to the prevalence of childhood obesity is estimated at 16%-40% in the United States, 10%-28% in Australia and Italy and 4%-18% in Great Britain, Sweden and The Netherlands. CONCLUSIONS: The contribution of TV advertising of foods and drinks to the prevalence of childhood obesity differs distinctly by country and is likely to be significant in some countries. PMID- 20018124 TI - Cannabis use and neuropsychological performance in healthy individuals and patients with schizophrenia. AB - BACKGROUND: The effects of cannabis use on neuropsychological indices that show characteristic disturbances in schizophrenia are unclear. The effect of cannabis use on these cognitive functions is of particular interest given the hypothesized association between cannabis use and schizophrenia. Therefore, this study aimed to examine the effects of cannabis use on attentional control, working memory and executive functioning, in both healthy individuals and patients with schizophrenia. METHOD: Neuropsychological performance was assessed in 36 cannabis users who were otherwise healthy, 35 healthy non-users, 22 cannabis-using patients with schizophrenia, and 49 non-using patients with schizophrenia. Participants were administered the Stroop task, the letter-number sequencing and spatial span subtests of the Wechsler Memory Scale, and the Wisconsin Card Sorting Test (WCST). RESULTS: Patients with schizophrenia (both cannabis users and non-users) showed significantly poorer performance across all neuropsychological tasks, relative to controls; however, there were no significant differences between schizophrenic cannabis users and schizophrenic non-users on any measures, with the exception of increased non-perseverative errors on the WCST in cannabis-using patients. Similarly, healthy cannabis users showed no significant differences from healthy non-users in any of the cognitive domains, with the exception of a schizophrenic-like increase in perseveration on the WCST. CONCLUSIONS: Amongst both healthy individuals and patients with schizophrenia there appears to be little difference in cognitive performance between cannabis users and non-users, suggesting that cannabis use has only subtle effects on the neurocognitive performance indices assessed here, which have been well established to be disturbed in schizophrenia. PMID- 20018125 TI - Temperamental and acute symptoms of borderline personality disorder: associations with normal personality traits and dynamic relations over time. AB - BACKGROUND: Recent research suggests the utility of distinguishing temperamental and acute symptoms of borderline personality disorder (BPD). Temperamental symptoms, such as chronic anger and odd thinking, remit relatively slowly and have been hypothesized to reflect a hyperbolic predisposition to emotional pain and negativistic cognitions, whereas acute symptoms, such as substance abuse and chaotic relationships, remit relatively quickly and have been hypothesized to represent the consequences of maladaptations to triggering environmental events. METHOD: The relationships of temperamental and acute BPD symptoms with normal personality traits and stability and dynamic associations over time across these symptom sets were tested in a 10-year longitudinal study of 362 patients with personality disorders. RESULTS: Temperamental symptoms were associated with high neuroticism, whereas acute symptoms were associated with low agreeableness. These symptoms had similar rank-order stabilities and relative changes in symptom sets were reciprocally linked in a cross-lagged path model suggesting dynamic associations between temperamental and acute symptoms over time. CONCLUSIONS: The distinction between temperamental and acute BPD symptoms is supported by differential relations of these symptom sets to normal personality traits. Moreover, these symptoms appear to be linked in a mutually reinforcing dynamic over time. This distinction should be kept in mind in future studies of the aetiology of BPD and in diagnostic and treatment considerations. PMID- 20018127 TI - Quantitative risk assessment to compare the risk of rabies entering the UK from Turkey via quarantine, the Pet Travel Scheme and the EU Pet Movement Policy. AB - Rabies was eradicated from the UK in 1922 through strict controls of dog movement and investigation of every incident of disease. Amendments were made to the UK quarantine laws and the Pet Travel Scheme (PETS) was subsequently introduced in 2000 for animals entering the UK from qualifying listed countries. European Regulation 998/2003 on the non-commercial movement of pet animals initiated the European Union Pet Movement Policy (EUPMP) in July 2004. The introduction of EUPMP harmonized the movement of pet animals within the EU (EUPMP(listed)) but raised the possibility of domestic animals entering the UK from a non-EU state where rabies is endemic (EUPMP(unlisted)). A quantitative risk assessment was developed to estimate the risk of rabies entering the UK from Turkey via companion animals that are incubating the disease and enter through PETS or EUPMP compared to quarantine. Specifically, the risk was assessed by estimating the annual probability of rabies entering the UK and the number of years between rabies entries for each scheme. The model identified that the probability of rabies entering the UK via the three schemes is highly dependent on compliance. If 100% compliance is assumed, PETS and EUPMP(unlisted) (at the current level of importation) present a lower risk than quarantine, i.e. the number of years between rabies entry is more than 170 721 years for PETS and 60 163 years for EUPMP(unlisted) compared to 41 851 years for quarantine (with 95% certainty). If less than 100% compliance is assumed, PETS and EUPMP(unlisted) (at the current level of importation) present a higher risk. In addition, EUPMP(listed) and EUPMP(unlisted) (at an increased level of importation) present a higher risk than quarantine or PETS at 100% compliance and at an uncertain level of compliance. PMID- 20018126 TI - Childhood adversity, adult stressful life events, and risk of past-year psychiatric disorder: a test of the stress sensitization hypothesis in a population-based sample of adults. AB - BACKGROUND: Childhood adversity (CA) is associated with adult mental disorders, but the mechanisms underlying this association remain inadequately understood. Stress sensitization, whereby CA increases vulnerability to mental disorders following adult stressful life events, has been proposed as a potential mechanism. We provide a test of the stress sensitization hypothesis in a national sample. METHOD: We investigated whether the association between past-year stressful life events and the 12-month prevalence of major depression, post traumatic stress disorder (PTSD), other anxiety disorders, and perceived stress varies according to exposure to CA. We used data from the National Epidemiological Survey of Alcohol and Related Conditions (NESARC) (n=34 653). RESULTS: Past-year stressful life events were associated with an increased risk of major depression, PTSD, anxiety disorders, and perceived stress. However, the magnitude of the increased risk varied according to respondents' history of CA. For example, past-year major stressors were associated with a 27.3% increase in the 12-month risk of depression among individuals with 3 CAs and a 14.8% increased risk among individuals without CAs. Stress sensitization effects were present for depression, PTSD, and other anxiety disorders in women and men, although gender differences were found in the threshold of past-year stress needed to trigger such effects. Stress sensitization was most evident among individuals with 3 CAs. CONCLUSIONS: CA is associated with increased vulnerability to the deleterious mental health effects of adult stressors in both men and women. High levels of CA may represent a general diathesis for multiple types of psychopathology that persists throughout the life course. PMID- 20018128 TI - A cost-utility analysis of antenatal screening to prevent congenital rubella syndrome. AB - In low vaccination coverage regions (LVR) in The Netherlands people often reject participation in the National Immunization Programme for religious reasons. During a rubella epidemic in 2004-2005, 32 pregnant women were notified with rubella, and 11 babies were born with defects related to maternal infection. This study presents a cost-utility analysis of a screening and vaccination programme for rubella focusing on three scenarios: (1) screening non-vaccinated pregnant women in LVR; (2) screening all pregnant women in LVR; (3) screening all non vaccinated pregnant women in The Netherlands (including pregnant first-generation non-Western immigrant women). Cost-utility was estimated over a 16-year period which included two rubella outbreaks. Observed complications from the 2004-2005 epidemic were used to estimate average cost savings and quality-adjusted life years (QALY) gained. The programme would be cost-effective (euro1100/QALY gained) when assuming an acceptability of vaccination of 20% in women belonging to orthodox protestant risk groups. PMID- 20018129 TI - Modelling multi-type transmission of pneumococcal carriage in Bangladeshi families. AB - The burden of pneumococcal carriage is largest in developing countries from which, however, detailed studies on pneumococcal transmission are missing. In this study we followed nasopharyngeal carriage in Bangladeshi infants (n=99) from birth, with 2-week sampling intervals until age 4 months, and monthly thereafter until age 1 year, and also their family members at the same intervals. We assessed the dependence of pneumococcal acquisition rates on age, serotype, serotype-specific exposure (i.e. transmission) and current state of carriage (yes/no). A statistical model of pneumococcal transmission, taking into account incompletely observed data, was applied to estimate rates of acquisition and clearance for a large number of serotypes at the same time. Serotypes that were common in the study population were more often acquired from the community than rarer serotypes. However, when conditioning on serotype-specific exposure within the family, transmission rates were similar between different serotypes. Exposure within families signified more than tenfold increase in the rate of acquisition. PMID- 20018130 TI - Surveillance for outbreaks of influenza-like illness in the institutionalized elderly. AB - Respiratory outbreaks are common in aged-care facilities (ACFs), are both underreported and frequently identified late, and are often associated with considerable burden of illness and death. There is emerging evidence that active surveillance coupled with early and systematic intervention can reduce this burden. Active surveillance for influenza-like illness and rapid diagnosis of influenza were established in 16 ACFs in Sydney, Australia, prior to the winter of 2006. A point-of-care influenza test and laboratory direct immunofluorescence tests for common respiratory viruses were used for diagnosis. We achieved early identification of seven respiratory disease outbreaks, two of which were caused by influenza. For the influenza outbreaks, antiviral treatment and prophylaxis were initiated 4-6 days from symptom onset in the primary case. A simple active surveillance system for influenza was successfully implemented and resulted in early detection of influenza and other respiratory disease outbreaks. This enabled earlier implementation of prevention and control measures and increased the potential effectiveness of anti-influenza chemoprophylaxis. PMID- 20018131 TI - Chlamydia pneumoniae infection is associated with elevated body mass index in young men. AB - Chlamydia pneumoniae infection is said to be associated with obesity. We studied the association between C. pneumoniae infection and inflammation and increased BMI in 891 Finnish military recruits. IgG seropositivity in arrival and departure serum samples during 6-12 months of military service was considered as persistence of antibodies and a possible indication of chronic infection. Persistently high C-reactive protein (CRP) level (elevated on arrival and departure) (OR 2.2, 95% CI 1.3-3.9), and persistent C. pneumoniae antibodies (OR 2.1, 95% CI 1.5-2.8) were significant risk factors for overweight (BMI 25 kg/m2). In addition, those who had persistent antibodies and persistently elevated CRP levels, or those who had either of them, had a significantly higher BMI (kg/m2) compared to those who had neither of them (25.8 vs. 24.6 vs. 23.5, respectively; P<0.001). These results provide new information about the association between possible chronic C. pneumoniae infection and obesity in young men. PMID- 20018132 TI - An investigation of women's attitudes towards fertility and China's family planning policy. AB - Reducing China's population has formerly been considered a good thing because of the perceived environmental and social risks of overpopulation, but it has recently become apparent that the resulting population decline may create problems that will become increasingly serious in the future. The results of a survey of 4600 women in nineteen Chinese provinces in 2005 indicated that young age, high income, high education level, urban location and good employment all decreased a woman's willingness to bear children. The risks created by declining fertility in these groups have been intensified by China's 'one child' family planning policy. However, as a result of current trends and China's policies, the country's population will continue to age, leading to social problems and difficulties for sustainable development both in China and around the world. Therefore, China's policy-makers must begin planning to adjust their policy by encouraging women to give birth to more than one child. PMID- 20018133 TI - Morbidity and mortality after surgery for congenital cardiac disease in the infant born with low weight. AB - OBJECTIVE: Low weight at birth is a risk factor for increased mortality in infants undergoing surgery for congenitally malformed hearts. There has been a trend towards performing surgery in patients early, and for amenable lesions, in a single stage rather than following initial palliative procedures. Our goal was to report on the current incidences of morbidities and mortality in infants born with low weight and undergoing surgery for congenital cardiac disease. METHODS: We made a retrospective review of the data from patients meeting our criterions for entry from July, 2000, through July, 2004. The criterions for inclusion were weight at birth less than or equal to 2500 grams, and congenital cardiac malformations requiring surgery during the initial hospitalization. A criterion for exclusion was isolated persistent patency of the arterial duct. We assessed preoperative, intraoperative, and postoperative variables. RESULTS: We found a total of 105 patients meeting the criterions for inclusion. The median weight at birth was 2130 grams, and median gestational age was 36 weeks. The most common morbidity identified was infections of the blood stream. Infections, and chronic lung disease, were associated with increased length of stay. Survival overall was 76%. Patients with hypoplastic left heart syndrome, or a variant thereof, had the lowest survival, of 62%. The needs for cardiopulmonary resuscitation, or extracorporeal membrane oxygenation, post-operatively were the only factors identified as independent risk factors for mortality. CONCLUSION: Patients undergoing surgery during infancy for congenital cardiac disease who are born with low weight have a higher mortality and morbidity than those born with normal weight. PMID- 20018134 TI - Evaluation of myocardial function using the Tei index in patients with Kawasaki disease. AB - Myocarditis is a well-recognized component of Kawasaki disease, with left ventricular dysfunction occurring in more than half of the patients during the acute phase of the disease. The purpose of our study was to evaluate myocardial function in these patients using the myocardial performance index, also known as the Tei index. In a prospective study, 14 patients underwent echocardiographic evaluation, first at the time of diagnosis of the disease, in its acute phase before treatment with intravenous immunoglobulin and then 2 weeks later after treatment with immunoglobulin. We assessed the Tei-index, the ejection fraction, shortening fraction, and the presence of valvar regurgitation, pericardial effusion, or coronary arterial involvement. As a control, we also assessed 22 healthy children, matched for age and sex with the study population. Of the patients, half had an increased left ventricular Tei-index in the acute phase, as compared with the data obtained after treatment, the index changing from 0.43 +/- 0.08 to 0.35 +/- 0.06 (p equal to 0.003). An increased index for the right ventricle was found in 5 patients (36%), values being 0.30 +/- 0.05 as opposed to 0.26 +/- 0.04 (p equal to 0.009). Of the patients, 5 (35.7%) also had decreased ejection fractions and proportional shortening fractions during the acute phase, confirming left ventricular dysfunction. We concluded that the Tei-index, which measures combined systolic and diastolic function, is a simple, sensitive, and accurate tool for estimating global myocardial dysfunction in patients with Kawasaki disease. PMID- 20018135 TI - Football fans and food: a case study of a football club in the English premier league. AB - OBJECTIVE: Although there is growing awareness of the impact of diet on health, little attention has been given to the food available in our sports stadia. We used a football club (Citygrene FC) - Citygrene is a fictional name - in the English Premier League as a case study to examine the attitudes of male and female football supporters to the food and drink available at their home stadium (Citygrene Stadium). DESIGN: The research design used five focus groups of male and female fans. The discourse was audiotaped, transcribed, coded and analysed for themes. SETTING: A football stadium in the English Premier League, England. SUBJECTS: The participants were season ticket holders drawn from two stands at Citygrene Stadium. RESULTS: The research showed a high level of dissatisfaction with the food and drink supplied. There were key differences in the views of the male and female participants in the focus groups, with the women more concerned about wider issues such as the lack of healthy food. Both men and women were aware of their role as consumers and felt that there was an opportunity for Citygrene to improve their catering profits, if they provided a better selection of food and drink and an improved service. CONCLUSIONS: The study shows that there is a demand for healthier food options (and a wider choice of food and drink in general), which may provide an economic opportunity for stadium and catering managers. In addition, a stadium may be considered a potential 'healthy setting', which can serve as a supportive environment for healthier food choices. PMID- 20018136 TI - Death in the United States, 2007. AB - KEY FINDINGS: Data from the National Vital Statistics System, Mortality In 2007, the age-adjusted death rate for the United States reached a record low of 760.3 per 100,000 population. Life expectancy at birth reached a record high of 77.9 years. States in the southeast region have higher death rates than those in other regions of the country. In 2007, the five leading causes of death were heart disease, cancer, stroke, chronic lower respiratory diseases, and accidents. These accounted for over 64 percent of all deaths in the United States. White females have the longest life expectancy (80.7 years), followed by black females (77.0 years). The gap in life expectancy between white persons and black persons declined by 35 percent between 1989 and 2007. The race differential was 4.6 years in 2007. PMID- 20018137 TI - Innocent pain: the experience of pain in metaphysics, phenomenology and neurophilosophy. PMID- 20018138 TI - Neurophilosophy and the distal hyperuranic world: Fred Previc's space of the gods (and of men). AB - Unlike the concept handed down by philosophy (and by the philosophy of science), which proposes a univocal, intellectualistic vision of space, Previc's studies have shown that the relationship between consciousness and space is instead managed by a number of modules, making it possible to categorise spatial interaction on four levels which correspond to the different ways in which we project our existence in different practical and cultural scenarios. What emerges as particularly important is the fourth level of the management of spatiality, the one that sees human consciousness dealing with the great distances - the heavens, the house of the gods. This module makes use of a neural pathway involving ventral sensory structures that also manage the relationship with religion and transcendence. But these are also the structures that are activated when populations are forced to set out in search of new lands - the quest for the eternal "Promised Land". In this study of infinite spaces, the path of the gods and of men coincide and they are seen to advance along it together. PMID- 20018139 TI - Chorea-ballismus in acute non-ketotic hyperglycaemia. AB - Diabetic patients during hyperglycaemic crises may present a rare syndrome characterised by a typical triad: unilateral involuntary movements (hemichoreahemiballism), radiological contralateral striatal abnormality, and rapid resolution of symptoms after glycae - mic correction. This study reports a series of patients showing less usual aspects and also discusses the pathophysiology of this clinical-radiological syndrome. We included in this study four patients presenting choreic or ballic involuntary movements and in whom aetiological assessment revealed frank non-ketotic hyperglycaemia, without other abnormalities that could explain the movement disorder. All the patients underwent CT or MR brain imaging. The typical triad was present in only one case. Less classical aspects were more frequently found: movement disorders revealed diabetes in two patients and one patient had generalised chorea and strictly normal neuroimaging. Correction of blood glucose was not sufficient to improve symptoms in two cases. In one, abnormal movements persisted despite treatment with tetrabenazine. The clinical, radiological and outcome spectrum of the syndrome of chorea-ballismus induced by non-ketotic hyperglycaemia is heterogeneous and not restricted to a typical triad. PMID- 20018140 TI - The severity of chronic cerebrospinal venous insufficiency in patients with multiple sclerosis is related to altered cerebrospinal fluid dynamics. AB - Chronic cerebrospinal venous insufficiency (CCSVI) is a vascular picture that shows a strong association with multiple sclerosis (MS). The aim of this study was to investigate the relationship between a Doppler cerebral venous hemodynamic insufficiency severity score (VHISS) and cerebrospinal fluid (CSF) flow dynamics in 16 patients presenting with CCSVI and relapsing-remitting MS (CCSVI-MS) and in eight healthy controls (HCs). The two groups (patients and controls) were evaluated using validated echo-Doppler and advanced 3T-MRI CSF flow measures. Compared with the HCs, the CCSVI-MS patients showed a significantly lower net CSF flow (p=0.027) which was highly associated with the VHISS (r=0.8280, r2=0.6855; p=0.0001). This study demonstrates that venous outflow disturbances in the form of CCSVI significantly impact on CSF pathophysiology in patients with MS. PMID- 20018141 TI - Interobserver and intraobserver agreement in the evaluation of CT perfusion in ischemic stroke. AB - Knowledge of interrater reliability in the evaluation of perfusion computed tomography (CTP) studies is very limited even though the method is widely used in the workup of acute stroke. The aims of this study were to estimate the inter- and intraobserver agreement in the evaluation of CTP data and to evaluate the feasibility of the method. The CTP data of 20 consecutive patients (50% were females) aged 68+/-11 years with different categories of acute ischemic stroke were included in this retrospective analysis. Perfusion studies were evaluated independently by six radiologists on two different occasions. The overall inter- and intraobserver agreement was substantial, showing a capital KA, Cyrillic value of 0.65 (95% confidence interval 0.39-0.91). The time required for the post processing and interpretation ranged from 37 to 460 seconds. Evaluation of manually post-processed CTP data according to the maximum slope model appears to be reliable. Experience and also a short training period increase the reliability of the method and reduce the time needed for delivery of the results to the treating clinician. PMID- 20018142 TI - Gait analysis and cerebral volumes in Down's syndrome. AB - The aim of this study was to look for a relationship between cerebral volumes computed using a voxel-based morphometry algorithm and walking patterns in individuals with Down's syndrome (DS), in order to investigate the origin of the motor problems in these subjects with a view to developing appropriate rehabilitation programmes. Nine children with DS underwent a gait analysis (GA) protocol that used a 3D motion analysis system, force plates and a video system, and magnetic resonance imaging (MRI). Analysis of GA graphs allowed a series of parameters to be defined and computed in order to quantify gait patterns. By combining some of the parameters it was possible to obtain a 3D description of gait in terms of distance from normal values. Finally, the results of cerebral volume analysis were compared with the gait patterns found. A strong relationship emerged between cerebellar vermis volume reduction and quality of gait and also between grey matter volume reduction of some cerebral areas and asymmetrical gait. An evaluation of high-level motor deficits, reflected in a lack or partial lack of proximal functions, is important in order to define a correct rehabilitation programme. PMID- 20018143 TI - Saccadic characteristics in autistic children. AB - Some studies suggest that individuals with autism present abnormal saccadic eye movements due to an altered strategy for exploration of the surrounding environment. In this study, potential early abnormalities of saccadic movements were explored in 14 male children with autism (5- to 12-year-olds) and in 20 age matched normal males. Only one patient showed clear abnormalities of the "main sequence"; all the other patients, although showing slight changes in saccadic eye movements, did not present classic deficits. Therefore our results did not confirm the presence of saccadic movement alterations in the early stage of autism. Nonetheless, tracts of saccadic initiation failure, continuous changes in saccadic velocity profiles, and instability of fixation were often observed in the autistic population. These findings could be the expression of an early brainstem impairment in autism. PMID- 20018145 TI - Expression of the three nitric oxide synthase isoforms and nitric oxide level in the rat heart during cold storage and blood reperfusion. AB - Maintenance of nitric oxide (NO) homeostasis is an important concept for myocardial protection. Here, we have investigated the NO pathway by analysing total nitrate concentration (NOx) and NO synthase (NOS) isoforms expression as well as the myocardial integrity by lactate dehydrogenase and creatine kinase contents in the rat heart graft arrested by CRMBM solution, submitted to 3 hr cold ischemia in the same solution and 24 hr blood reperfusion following heterotopic abdominal heart transplantation. NOx level was similar to baseline value after ischemia and significantly increased after 24 hr reperfusion. NOS isoforms expression was highly modulated after cold ischemia followed by blood reperfusion. Endothelial NOS expression was decreased after ischemia but restored after 24 hr reperfusion. Neuronal NOS expression was drastically decreased after ischemia and 24 hr reperfusion. Inducible NOS protein was present only after 24 hr reperfusion. Cold ischemia induced a severe loss of creatine kinase without any modification after blood reperfusion. In conclusion, we show here that CRMBM solution did not increase NO production during ischemia but induced an enhanced synthesis of NO during reperfusion which may be related to restoration of endothelial NOS expression and/or induction of inducible NOS expression. PMID- 20018144 TI - Validation of the DYMUS questionnaire for the assessment of dysphagia in multiple sclerosis. AB - Swallowing problems can complicate the course of multiple sclerosis (MS). However, no validated questionnaire for the assessment of dysphagia in MS is currently available. We previously developed a 10-item DYsphagia in Multiple Sclerosis questionnaire (DYMUS). In the present study, this questionnaire was submitted to a validation process. Thirteen Italian MS centres took part in this research in which DYMUS was administered to 1734 consecutive MS patients during routine checkups outside relapse. The questionnaire showed very good internal consistency (Cronbach's alpha = 0.914). It was then subdivided into two subscales, both of which also showed very good internal consistency: Cronbach's alpha was 0.885 for the 'dysphagia for solids' subscale and 0.864 for the 'dysphagia for liquids' subscale. The DYMUS questionnaire was found to be an easy and reliable tool for detecting dysphagia and also for the preliminary selection of patients requiring more specific instrumental analyses, and those suitable for aspiration prevention programmes. PMID- 20018146 TI - Are adverse effects incorporated in economic models? An initial review of current practice. AB - OBJECTIVES: To identify methodological research on the incorporation of adverse effects in economic models and to review current practice. DATA SOURCES: Major electronic databases (Cochrane Methodology Register, Health Economic Evaluations Database, NHS Economic Evaluation Database, EconLit, EMBASE, Health Management Information Consortium, IDEAS, MEDLINE and Science Citation Index) were searched from inception to September 2007. Health technology assessment (HTA) reports commissioned by the National Institute for Health Research (NIHR) HTA programme and published between 2004 and 2007 were also reviewed. REVIEW METHODS: The reviews of methodological research on the inclusion of adverse effects in decision models and of current practice were carried out according to standard methods. Data were summarised in a narrative synthesis. RESULTS: Of the 719 potentially relevant references in the methodological research review, five met the inclusion criteria; however, they contained little information of direct relevance to the incorporation of adverse effects in models. Of the 194 HTA monographs published from 2004 to 2007, 80 were reviewed, covering a range of research and therapeutic areas. In total, 85% of the reports included adverse effects in the clinical effectiveness review and 54% of the decision models included adverse effects in the model; 49% included adverse effects in the clinical review and model. The link between adverse effects in the clinical review and model was generally weak; only 3/80 (< 4%) used the results of a meta analysis from the systematic review of clinical effectiveness and none used only data from the review without further manipulation. Of the models including adverse effects, 67% used a clinical adverse effects parameter, 79% used a cost of adverse effects parameter, 86% used one of these and 60% used both. Most models (83%) used utilities, but only two (2.5%) used solely utilities to incorporate adverse effects and were explicit that the utility captured relevant adverse effects; 53% of those models that included utilities derived them from patients on treatment and could therefore be interpreted as capturing adverse effects. In total, 30% of the models that included adverse effects used withdrawals related to drug toxicity and therefore might be interpreted as using withdrawals to capture adverse effects, but this was explicitly stated in only three reports. Of the 37 models that did not include adverse effects, 18 provided justification for this omission, most commonly lack of data; 19 appeared to make no explicit consideration of adverse effects in the model. CONCLUSIONS: There is an implicit assumption within modelling guidance that adverse effects are very important but there is a lack of clarity regarding how they should be dealt with and considered in modelling. In many cases a lack of clear reporting in the HTAs made it extremely difficult to ascertain what had actually been carried out in consideration of adverse effects. The main recommendation is for much clearer and explicit reporting of adverse effects, or their exclusion, in decision models and for explicit recognition in future guidelines that 'all relevant outcomes' should include some consideration of adverse events. PMID- 20018147 TI - [Ugeskrift for Laeger on the net]. PMID- 20018148 TI - [Oram's sign for hip or pelvic disorder]. AB - INTRODUCTION: The so-called Oram's sign states that the penis points to the side of the disorder on standard antero-posterior (AP) x-rays of the pelvis. The present study is a scientific evaluation of the sign in a relevant clinical context. MATERIAL AND METHODS: This is a retrospective study based on a blinded reading by four males and one female doctor of AP x-rays of the pelvis from 51 male patients with hip and pelvic disorders and 11 x-rays without pathology from an additional nine males and two females. The sensitivity and specificity and the positive and negative likelihood ratios were calculated together with the Kappa values for intra- and interobserver agreement. RESULTS: The sensitivity of Oram's sign for the individual reader varied between 40.3% and 25.8%. The overall sensitivity and specificity for all readers were 55% and 14%, respectively. The test performance did not show any significant variation between the male readers and the female reader. The sensitivity of Oram's sign in relation to the various pathologic findings varied between 32.1% for femoral neck fractures, 66.6% for osteoarthritis of the hip and 88.8% for pelvic fracture. Oram's sign could not be defined in 16.3% of the cases. The intra- and interobserver agreement was poor with relatively low Kappa values. CONCLUSION: Oram's sign was not useful in this study on the diagnosis of hip fractures mainly due to a low sensitivity and too low observer agreement. PMID- 20018149 TI - [The significance of appearance in physician-nurse collaboration]. AB - INTRODUCTION: According to nurses' assessment, physician-nurse collaboration is problematic. The aim of the study was to investigate whether nurses believe physicians' appearances is significant for their ability to collaborate. MATERIAL AND METHODS: This is a single-blinded, quasi-experimental intervention study. All physicians included will undergo a total plastic surgical body makeover. Half of the included physicians are to be operated with a view to enhance their physical features, half to weaken their physical features. Multifactorial ANOVA analysis will be used to secure the division. The Jefferson Scale of Attitudes Toward Physician-Nurse Collaboration will be used for baseline and follow-up study of the nurses' assessment. RESULTS: Due to ethical considerations, researchers had difficulties finding surgeons prepared to perform procedures aiming at weakening the physicians' physical features. Furthermore, it proved difficult to find physicians willing to undergo surgery with a view to weaken their physical features. Consequently, the study was never realized. Additionally, the study experienced difficulties raising the funding needed to cover surgery costs for all included physicians. CONCLUSION: We conclude that physicians often have difficulties collaborating with nurses, regardless of their appearance. It seems that physicians are not interested in improving the collaboration with nurses, and further studies are therefore needed to improve physician-nurse collaboration. We recommend in-depth interviews to achieve harmonic interaction leading to a prolific and close future collaboration. PMID- 20018150 TI - [On the effect of shotgun shot. A crime scene investigation]. AB - Rifle contra shotgun shots are considered in relation to an X-mas goose and shooting accident. Depending on the size of shots, material and velocity, the effect in the body may vary considerably. Myocardial infarction, secondary to accidental shotgun shot, may be caused by 1) thrombosis secondary to intima lesion, 2) stenosis, oedema and haematoma in relation to myocardial lesion; and, in addition, 3) the cumulative energy of many shots may give myocardial contusion with infarction. Pathotechnical and pathophysiological analyses are required to reveal complicated causes and effects and unusual disease events. PMID- 20018151 TI - [Folie a deux]. AB - Folie a deux or induced psychotic disorder is a rare psychiatric illness. The essential feature is a psychotic disorder shared by two emotionally close persons, usually close relatives. The disorder is characterized by a primary psychotic person who gradually imposes her delusional beliefs on the other person. The disorder usually resolves in the more passive person when the two are separated (e.g. by admittance of the primary psychotic person). This case report describes a shared delusional belief between two friends. PMID- 20018152 TI - [Goodbye to half of tomorrow's physicians]. AB - INTRODUCTION: Danish citizenship can only be obtained by passing a citizenship test. The test was introduced in December 2005 which made it more difficult to obtain Danish citizenship, and revised in 2008 which made the process even harder. We present a study which tests how many Danish and Swedish medical students enrolled at the University of Copenhagen would be able to obtain a Danish citizenship. MATERIAL AND METHODS: The test comprised 40 questions regarding Danish democracy, history and politics. The questions were taken from the Ministry of Refugee and Immigration's pool of questions. Medical students at the University of Copenhagen were invited to participate in the study. Participation was voluntary. The tests were revised blinded and anonymously. The results were analyzed using Fisher's exact test and Mann-Whitney U-test. RESULTS: We enrolled 202 medical students (139 women and 63 men). Among these participants, 168 were Danish and 34 Swedish medical students studying in Copenhagen. A total of 49% of the students passed the test. 58% of the Danish students and 6% of the Swedish students passed the test (p < 0.001). The Danish students had a median of eight errors (range: 1-18) and Swedish students had a median of 15 errors (range: 5-25) (p < 0.001). CONCLUSION: We suggest an export agreement with countries like Poland and India comprising the 51% students who failed the citizenship test as part of a package solution, where Denmark in return receives competent physicians of said nationalities. We also suggest that the contents of the test for citizenship be revised to achieve a better assessment of the applicant's ability to integrate in the Danish society. PMID- 20018153 TI - [Life just is not fair - and statistics do not make it any better]. AB - INTRODUCTION: We have all tried it: it is pouring down with rain, we reach for the keys, and choose the wrong one. The mathematical probability for choosing the correct key - out of two - is 50%. During the 1990s, candy.scient. Jarle Gundersson (JG) proposed a mathematically unexplainable factor of uncertainty. He used the above example with the keys and concluded that the real, or observed, probability for choosing the correct key was 5-10%. The discrepancy between the mathematical and real probability was, according to JG, caused by The Awfulness of Being Concept. In this article, we present the results from a study and demonstrate the difference between the mathematical and the observed probability of success in two different scenarios (winning a coin toss and choosing the correct key in the first attempt). MATERIAL AND METHODS: Questionnaire survey performed at the Regional Hospital of Esbjerg 18-19 September 2009 using staff as questionnaire respondents. RESULTS: We found a discrepancy between the mathematical and observed probability in both scenarios, 64% and 68% of the cases in the two scenarios, respectively. We also found that there is a negative correlation between age and the probability that one will win a coin toss, but a positive correlation between age and choosing the correct key. The results lacked any statistical significance. CONCLUSION: We hypothesised that you will always loose in a coin toss and never choose the correct key when there are two choices. The study demonstrated that this was incorrect, but there was a difference between the mathematical and the observed probabilities in most of the cases. PMID- 20018154 TI - [Evolution of the ear]. AB - The evolution of the ear may be traced through transitional fossils, comparative anatomy and embryology. The organ of hearing evolved from receptors of the vestibulary organ of fish. The tympanic ear developed in amphibians at the transition to terrestrial life, and the hyomandibula was isolated as the first middle ear bone, the columella stapes. Reptile jaw bones, quadratum and articulare, transformed to malleus and incus in mammals. With selective advantages during the evolution, an increasing structural complexity of the ear accompanied improved sound transmission and reception. PMID- 20018155 TI - [A new scientific source of bias: SILLY bias. Analysis of citations of BMJ's Christmas articles]. AB - We analysed the scientific impact of systematic reviews and randomised trials published in the BMJ Christmas issues 1997-2006. The articles were mostly interpreted correctly as humorous, but the humorous dimension was overlooked with surprising ease. The result from one ironic-absurd trial on the effect of retroactive remote intercessory prayer for patients already dead or dismissed was taken at face value in 12/36 of the citing articles, and mortality data was unconditionally included in three systematic reviews. Thus, we document a new type of bias in medical research: Serious Idiopathic Loss of Ludic ironY (SILLY) bias, both in citation practices and in metaanalyses. PMID- 20018156 TI - [Diagnostics and treatment of pruritus ani]. AB - Pruritus ani is defined as an intense itching sensation affecting the perianal skin. The condition affects up to 5% of the population, and occurs four times more commonly in men than in women. Pruritus ani occurs in a primary form caused by overt but insufficient cleansing of the perianal skin, and in a secondary form caused by underlying pathology, often of proctological or dermatological/infectious origin. Causes and treatment are reviewed, and it is concluded that the condition, even in its chronic refractory form, has a favourable prognosis. PMID- 20018157 TI - [Munchhausen's syndrome--a diagnosis to remember]. AB - We hereby report a case of self mutilation against the right upper extremity in a 26-year-old male. The condition started after a minor trauma. After a few days the diagnosis compartment syndrome was suspected and a fasciectomy of the forearm was performed. The patient subsequently claimed that he had experienced further symptoms. Despite countless investigations, it was not possible to identify the course of the patient's symptoms. It has become clear that the patient suffers from >>Munchhausen's syndrome<<. PMID- 20018158 TI - Solubilization and functional reconstitution of human neuropeptide FF2 receptors. AB - Neuropeptide FF (NPFF, FLFQPQRFamide) receptors modulate endogenous opioid functions. Here, we report the solubilization of the human NPFF(2) receptor expressed in Chinese hamster ovary (CHO) cells by the zwitterionic detergent Chaps. Chaps solubilization resulted in the abolishment of specific agonist binding activity, which was restored by a polyethylene glycol (PEG) precipitation method. Reincorporation after the precipitation step into liposomes made of endogenous lipids issued from CHO membranes or exogenous lipids significantly enhanced the specific agonist binding activity and G-protein coupling. This method of solubilization and lipid reconstitution could be useful for studies of NPFF receptors. PMID- 20018159 TI - Use of thermal melt curves to assess the quality of enzyme preparations. AB - This study sought to determine whether the quality of enzyme preparations can be determined from their melting curves, which may easily be obtained using a fluorescent probe and a standard reverse transcription-polymerase chain reaction (RT-PCR) machine. Thermal melt data on 31 recombinant enzymes from Plasmodium parasites were acquired by incrementally heating them to 90 degrees C and measuring unfolding with a fluorescent dye. Activity assays specific to each enzyme were also performed. Four of the enzymes were denatured to varying degrees with heat and sodium dodecyl sulfate (SDS) prior to the thermal melt and activity assays. In general, melting curve quality was correlated with enzyme activity; enzymes with high-quality curves were found almost uniformly to be active, whereas those with lower quality curves were more varied in their catalytic performance. Inspection of melting curves of bovine xanthine oxidase and Entamoeba histolytica cysteine protease 1 allowed active stocks to be distinguished from inactive stocks, implying that a relationship between melting curve quality and activity persists over a wide range of experimental conditions and species. Our data suggest that melting curves can help to distinguish properly folded proteins from denatured ones and, therefore, may be useful in selecting stocks for further study and in optimizing purification procedures for specific proteins. PMID- 20018160 TI - Impact of phospholipid bilayer saturation on amyloid-beta protein aggregation intermediate growth: a quartz crystal microbalance analysis. AB - Evidence that membrane-associated amyloid aggregate growth can impart membrane damage represents one possible mechanism for the neurodegeneration associated with deposited amyloid-beta protein (Abeta) aggregates in the brains of Alzheimer's disease (AD) patients. This potential pathogenic event necessitates an understanding of the impact that cellular membrane composition may have on Abeta aggregate growth. In the current study, a quartz crystal microbalance (QCM) was employed to examine the growth of Abeta(1-40) aggregation intermediates on supported phospholipid bilayers (SPBs) assembled at the crystal surface. These surface-specific measurements illustrate that zwitterionic SPBs selectively bind aggregated but not monomeric protein, and these bound aggregates are capable of supporting nonsaturable reversible growth via monomer addition. Growth-capable Abeta(1-40) aggregation intermediates more readily bind SPBs composed of phospholipids with a greater degree of carbon saturation. Furthermore, kinetic analysis afforded by the quantitative real-time QCM measurements reveals that SPBs with greater saturation also better support the growth of bound Abeta(1-40) aggregation intermediates as a result of the slower dissociation of bound monomer rather than more efficient recognition between aggregate and monomeric protein. These findings correlate with epidemiological and experimental evidence that links increased dietary intake of polyunsaturated fatty acids to a reduced risk of AD. PMID- 20018161 TI - Analysis of branched-chain alpha-keto acid dehydrogenase complex activity in rat tissues using alpha-keto[1-13C]isocaproate as substrate. AB - We have developed a highly sensitive nonradioisotope method for the measurement of branched-chain alpha-keto acid dehydrogenase complex (BCKDC) activity using alpha-keto[1-(13)C]isocaproate ([1-(13)C]KIC) as substrate. The enzyme reaction was performed in a 10-ml test tube. After incubation of the reaction mixture (1 ml), the reaction was terminated by acidification, followed by the addition of internal standard (3.5 micromol K(2)CO(3)) for calculation of (13)CO(2). The (13)CO(2) released to the gas phase in the test tube was calculated from the molar ratio of (13)CO(2)/(12)CO(2) assayed by gas chromatography isotope ratio mass spectrometry. A distinct linear relationship between (13)CO(2) production rate and BCKDC activity was obtained when using purified BCKDC. The detection limit of (13)CO(2) in the assay method was at least 0.167 nmol, which is similar to that in the previously reported radiochemical method. The enzyme assay was found to be linear with respect to reaction time and quantity of enzyme used up to approximately 60 nmol (13)CO(2) produced. We measured BCKDC activities in several rat tissues, including skeletal muscle. The activities of BCKDC obtained are comparable to those reported previously. These results suggest that the novel BCKDC assay method using [1-(13)C]KIC is useful for measurement in tissues with low BCKDC activity such as skeletal muscle. PMID- 20018162 TI - Application of glutaraldehyde to in-cell Western assay for normalization. AB - Normalization is essential to the in-cell Western (ICW) assay, a near-infrared immunocytoblot for protein analysis. Here we report that cells reacted with glutaraldehyde fluoresced in the near-infrared region of the spectrum, and the intensity of fluorescence was directly proportional to cell number over a range from 3125 to 100,000 cells per well. We took advantage of this property to develop a method for quantification of cells, and applied it to the ICW assay for normalization. The application of glutaraldehyde may make the ICW assay more popular due to the reduced cost and simplified procedure. PMID- 20018163 TI - A fluorescent assay suitable for inhibitor screening and vanin tissue quantification. AB - Vanin-1 is a pantetheinase that catalyzes the hydrolysis of pantetheine to produce pantothenic acid (vitamin B5) and cysteamine. Reported here is a highly sensitive fluorescent assay using a novel fluorescently labeled pantothenate derivative. The assay has been used for characterization of a soluble version of human vanin-1 recombinant protein, identification and characterization of hits from high-throughput screening (HTS), and quantification of vanin pantothenase activity in cell lines and tissues. Under optimized assay conditions, we quantified vanin pantothenase activity in tissue lysate and found low activity in lung and liver but high activity in kidney. We demonstrated that the purified recombinant vanin-1 consisting of the extracellular portion without the glycosylphosphatidylinositol (GPI) linker was highly active with an apparent K(m) of 28 microM for pantothenate-7-amino-4-methylcoumarin (pantothenate-AMC), which was converted to pantothenic acid and AMC based on liquid chromatography-mass spectrometry (LC-MS) analysis. The assay also performed well in a 384-well microplate format under initial rate conditions (10% conversion) with a signal-to background ratio (S/B) of 7 and a Z factor of 0.75. Preliminary screening of a library of 1280 pharmaceutically active compounds identified inhibitors with novel chemical scaffolds. This assay will be a powerful tool for target validation and drug lead identification and characterization. PMID- 20018165 TI - ATP-binding cassette transporters as pitfalls in selection of transgenic cells. AB - Puromycin, hygromycin, and geneticin (G418) are antibiotics frequently used to select genetically engineered eukaryotic cells after transfection or transduction. Because intrinsic or acquired high expression of ATP-binding cassette (ABC) transporters, such as P-glycoprotein (Pgp/ABCB1) and multidrug resistance-associated proteins (MRP/ABCC1), can hamper efficient selection, it is important to know whether these antibiotics are substrates and/or inducers of efflux transporters. Therefore, we investigated the influence of these antibiotics on drug transporter expression by quantitative real-time polymerase chain reaction in the induction model cell line LS180. Moreover, we assessed whether ABC transporters influence the growth inhibitory effects of these antibiotics by proliferation assays using Madin-Darby canine kidney II (MDCKII) cells overexpressing the particular transporter. The results obtained indicate that puromycin and G418 are substrates of several ABC transporters, mainly Pgp/ABCB1. In contrast, hygromycin seems to be no good substrate for any of the ABC transporters investigated. Puromycin induced ABCC1/MRP1, whereas G418 suppressed ABCB1/Pgp, at the messenger RNA (mRNA) level. In contrast, hygromycin had no effect on ABC transporter mRNA expressions. In conclusion, this study emphasizes the significance of ABC transporters for the efficacy of selection processes. Consciousness of the results is supposed to guide the molecular biologist to the right choice of adequate experimental conditions for successful selection of genetically engineered eukaryotic cells. PMID- 20018166 TI - Nuclear magnetic resonance quantification for monitoring heparosan K5 capsular polysaccharide production. AB - Traditional chromatographic quantification methods for heparosan produced from the Escherichia coli K5 strain rely on extensive purification requiring laborious sample preparation. These methods are time-consuming, often resulting in sample loss during purification, and thus might not accurately reflect the amount of heparosan in the original mixture. A simple, sensitive (1)H nuclear magnetic resonance (NMR) quantification method that directly quantifies heparosan K5 polysaccharide present in E. coli fermentation supernatant is described. PMID- 20018164 TI - Identification of inhibitors of vacuolar proton-translocating ATPase pumps in yeast by high-throughput screening flow cytometry. AB - Fluorescence intensity of the pH-sensitive carboxyfluorescein derivative 2,7 bis(2-carboxyethyl)-5(6)-carboxyfluorescein (BCECF) was monitored by high throughput flow cytometry in living yeast cells. We measured fluorescence intensity of BCECF trapped in yeast vacuoles, acidic compartments equivalent to lysosomes where vacuolar proton-translocating ATPases (V-ATPases) are abundant. Because V-ATPases maintain a low pH in the vacuolar lumen, V-ATPase inhibition by concanamycin A alkalinized the vacuole and increased BCECF fluorescence. Likewise, V-ATPase-deficient mutant cells had greater fluorescence intensity than wild-type cells. Thus, we detected an increase of fluorescence intensity after short- and long-term inhibition of V-ATPase function. We used yeast cells loaded with BCECF to screen a small chemical library of structurally diverse compounds to identify V-ATPase inhibitors. One compound, disulfiram, enhanced BCECF fluorescence intensity (although to a degree beyond that anticipated for pH changes alone in the mutant cells). Once confirmed by dose-response assays (EC(50)=26 microM), we verified V-ATPase inhibition by disulfiram in secondary assays that measured ATP hydrolysis in vacuolar membranes. The inhibitory action of disulfiram against V-ATPase pumps revealed a novel effect previously unknown for this compound. Because V-ATPases are highly conserved, new inhibitors identified could be used as research and therapeutic tools in cancer, viral infections, and other diseases where V-ATPases are involved. PMID- 20018167 TI - Discovery and characterization of [3H]8-OH-DPAT binding to HeLaS3 cells. AB - Some G protein-coupled receptors (GPCRs) have functional links to cancer biology, yet the manifestation of GPCRs in tumor types is little studied to date. Using a battery of radioligand binding assays, we sought to characterize GPCR recognition binding sites on HeLaS3 tumor cells. High levels of binding of the selective serotonin 5-HT(1A) receptor agonist [3H]8-OH-DPAT were observed in these cells. Saturation and homologous competition experiments indicated that [3H]8-OH-DPAT bound different populations of high- and low-affinity sites. In competition experiments, several serotonergic compounds displaced [3H]8-OH-DPAT binding with low potency from its high-affinity binding sites, suggesting that low-affinity binding is the predominant mode of binding. A variety of drugs targeting different classes of receptors did not affect [3H]8-OH-DPAT binding. These observations may help elucidate the pathophysiological and functional relevance of 5-HT receptors in tumor cells and link GPCRs and tumorigenic mechanisms to pharmacological and chemotherapeutic paradigms. PMID- 20018168 TI - Mild thermotolerance induced at 40 degrees C increases antioxidants and protects HeLa cells against mitochondrial apoptosis induced by hydrogen peroxide: Role of p53. AB - Exposure of cells to mild temperatures (40 degrees C) induces thermotolerance, which renders cells resistant to subsequent toxic insults. Thermotolerance is usually associated with accumulation of heat shock proteins. This study determines whether mild thermotolerance (40 degrees C, 3h) can induce other defense proteins (e.g. antioxidants, anti-apoptosis proteins), and protect HeLa cells against apoptosis triggered by H(2)O(2). Protein expression and enzymatic activity of MnSOD and catalase were increased in thermotolerant cells, as well as intracellular glutathione levels and gamma-glutamylcysteine synthetase expression. Furthermore, levels of reactive oxygen species (ROS) were increased in thermotolerant cells, which caused mitochondrial membrane hyperpolarisation. Mild thermotolerance inhibited activation of the mitochondrial cascade of apoptosis by H(2)O(2). This entailed inhibition of mitochondrial Bax translocation, mitochondrial membrane depolarisation, cytochrome c release, activation of caspases-9/-3 and chromatin condensation. Thermotolerance inhibited H(2)O(2)-induced caspase-independent apoptosis involving apoptosis-inducing factor, and activation of p53 and increased expression of its target protein PUMA. Thermotolerance induced at mild physiological temperatures protects cells against both caspase-dependent and caspase-independent apoptosis triggered by oxidative stress. PMID- 20018169 TI - (-)-Epicatechin enhances the chlorinating activity of human myeloperoxidase. AB - The heme-containing enzyme myeloperoxidase (MPO) accumulates at inflammatory sites and is able to catalyse one- and two-electron oxidation reactions. Here it is shown that (-)-epicatechin, which is known to have numerous beneficial health effects, in low micromolar concentration enhances the degradation of monochlorodimedon (MCD) or the chlorination of taurine in a concentration dependent bell-shaped manner whereas at higher concentrations it sufficiently suppresses the release of hypochlorous acid. Presented reaction mechanisms demonstrate the efficiency of micromolar concentrations of the flavan-3-ol in overcoming the accumulation of compound II that does not participate in the chlorination cycle. In case of MCD the mechanism is more complicated since it also acts as peroxidase substrate with very different reactivity towards compound I (3 x 10(5) M(-1) s(-1)) and compound II (8.8M(-1)s(-1)) at pH 7. By affecting the chlorinating activity of myeloperoxidase (-)-epicatechin may participate in regulation of immune responses at inflammatory sites. PMID- 20018170 TI - Methyl-beta-cyclodextrin restores the structure and function of pulmonary surfactant films impaired by cholesterol. AB - Pulmonary surfactant, a defined mixture of lipids and proteins, imparts very low surface tension to the lung-air interface by forming an incompressible film. In acute respiratory distress syndrome and other respiratory conditions, this function is impaired by a number of factors, among which is an increase of cholesterol in surfactant. The current study shows in vitro that cholesterol can be extracted from surfactant and function subsequently restored to dysfunctional surfactant films in a dose-dependent manner by methyl-beta-cyclodextrin (MbetaCD). Bovine lipid extract surfactant was supplemented with cholesterol to serve as a model of dysfunctional surfactant. Likewise, when cholesterol in a complex with MbetaCD ("water-soluble cholesterol") was added in aqueous solution, surfactant films were rendered dysfunctional. Atomic force microscopy showed recovery of function by MbetaCD is accompanied by the re-establishment of the native film structure of a lipid monolayer with scattered areas of lipid bilayer stacks, whereas dysfunctional films lacked bilayers. The current study expands upon a recent perspective of surfactant inactivation in disease and suggests a potential treatment. PMID- 20018171 TI - Sphingomyelin-rich domains are sites of lysenin oligomerization: implications for raft studies. AB - Lysenin is a self-assembling, pore-forming toxin which specifically recognizes sphingomyelin. Mutation of tryptophan 20 abolishes lysenin oligomerization and cytolytic activity. We studied the interaction of lysenin WT and W20A with sphingomyelin in membranes of various lipid compositions which, according to atomic force microscopy studies, generated either homo- or heterogeneous sphingomyelin distribution. Liposomes composed of SM/DOPC, SM/DOPC/cholesterol and SM/DPPC/cholesterol could bind the highest amounts of GST-lysenin WT, as shown by surface plasmon resonance analysis. These lipid compositions enhanced the release of carboxyfluorescein from liposomes induced by lysenin WT, pointing to the importance of heterogeneous sphingomyelin distribution for lysenin WT binding and oligomerization. Lysenin W20A bound more weakly to sphingomyelin containing liposomes than did lysenin WT. The same amounts of lysenin W20A bound to sphingomyelin mixed with either DOPC or DPPC, indicating that the binding was not affected by sphingomyelin distribution in the membranes. The mutant lysenin had a limited ability to penetrate hydrophobic region of the membrane as indicated by measurements of surface pressure changes. When applied to detect sphingomyelin on the cell surface, lysenin W20A formed large conglomerates on the membrane, different from small and regular clusters of lysenin WT. Only lysenin WT recognized sphingomyelin pool affected by formation of raft-based signaling platforms. During fractionation of Triton X-100 cell lysates, SDS-resistant oligomers of lysenin WT associated with membrane fragments insoluble in Triton X 100 while monomers of lysenin W20A partitioned to Triton X-100-soluble membrane fractions. Altogether, the data suggest that oligomerization of lysenin WT is a prerequisite for its docking in raft-related domains. PMID- 20018172 TI - Hypoxia-induced decoy receptor 2 gene expression is regulated via a hypoxia inducible factor 1alpha-mediated mechanism. AB - Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) is a promising candidate for anti-tumor therapy because of its high selectivity towards cancer cells. TRAIL has four major distinct receptors: DR4 and DR5 can recruit Fas associated death domain protein to induce extrinsic death signal, while DcR1 and DcR2 are decoy receptors that can neutralize TRAIL toxicity by binding to TRAIL. Hypoxia is an important feature of solid tumors that renders tumor cells resistant to some chemotherapeutic agents, including TRAIL, and we therefore investigated the role of hypoxia in TRAIL receptor expression in human colon cancer cells. Hypoxia upregulated DcR2 protein expression in five different human colon cancer cell lines (HCT116, HT29, SW480, SW620, and WiDr). Flow cytometry analysis indicated that the increased DcR2 protein was expressed on the cell surface membrane. In contrast, hypoxia had no effect on DR4, DR5, or DcR1 protein levels. RT-PCR analysis suggested that this protein increase was the result of DcR2 gene transcription. Transcription factors were investigated using p53-null cells, pharmacological inhibitors, and a small interfering RNA approach. Our results demonstrated that hypoxia-inducible factor 1alpha played a crucial role in regulating the transcription of DcR2, but that neither p53 nor NF-kappaB contributed to this regulation. Moreover, TRAIL-induced, but not agonistic DR5 antibody-induced cell death was attenuated under hypoxic conditions. These results suggest that increased DcR2 protein levels might play a role in TRAIL resistance in solid tumors. PMID- 20018173 TI - Construction of a bacterial artificial chromosome library for the Rongchang pig breed and its use for the identification of genes involved in intramuscular fat deposition. AB - In a search for genes affecting intramuscular fat deposition, we constructed a bacterial artificial chromosome (BAC) library for the whole genome of Rongchang pig, a domestic Chinese swine breed. The library consisted of approximately 192,000 clones, with an averaged insert size of 116 kb. Frequency of non-insert clone of the BAC library was no higher than 1.8%, based on estimation of 220 BAC clones randomly selected. We estimated the coverage of the library to be more than seven porcine genome equivalents. Subsequent screening of the BAC library with a three-step PCR procedure resulted in identification of seven candidate genes that were potentially involved in intramuscular fat deposition. The number of positive BAC clones ranged from 2 to 4 for each of the seven genes. One positive clone, containing the lipin1 gene, was fully sequenced by shotgun method to generate 118,041 bp porcine genomic sequences. The BAC clone contained complete DNA sequence of porcine lipin1 gene including all the exons and introns. Our results indicate that this BAC library is a useful tool for gene identification and help to serve as an important resource for future porcine genomic study. PMID- 20018174 TI - Hyperglycemia induces elevated expression of thyroid hormone binding protein in vivo in kidney and heart and in vitro in mesangial cells. AB - During a search for glucose-regulated abundant mRNAs in the diabetic rat kidney, we cloned thyroid hormone binding protein (THBP), also known as mu-crystallin or CRYM. The aim of this study was to investigate the effect of hyperglycemia/high glucose on the expression of THBP. THBP mRNA copy numbers were determined in kidneys and hearts of diabetic GK rats vs normoglycemic Wistar rats, and in human mesangial cells (HMCs) exposed to high glucose using real-time qPCR, and THBP protein levels were measured by Western blotting and immunofluorescence. Intracellular ROS was measured in THBP transfected cells using DCF fluorescence. Hyperglycemia significantly increased THBP mRNA in GK rat kidneys (326+/-50 vs 147+/-54, p<0.05), and hearts (1583+/-277 vs 191+/-63, p<0.05). Moreover, the levels of THBP mRNA increased with age and hyperglycemia in GK rat kidneys, whereas in normoglycemic Wistar rat kidneys there was a decline with age. High glucose significantly increased THBP mRNA (92+/-37 vs 18+/-4, p<0.005), and protein in HMCs. The expression of THBP as a fusion protein in transfected HMCs resulted in reduction of glucose-induced intracellular ROS. We have shown that THBP mRNA is increased in diabetic kidney and heart, is regulated by high glucose in renal cells, and appears to attenuate glucose-induced intracellular ROS. These data suggest that THBP may be involved in the cellular pathways activated in response to glucose. This is the first report linking hyperglycemia with THBP and suggests that the role of THBP in diabetic complications should be further investigated. PMID- 20018175 TI - Gene-wide histone acetylation at the yeast INO1 requires the transcriptional activator Ino2p. AB - The relationship between histone acetylation and transcriptional activation at the yeast INO1 gene was addressed. INO1 encodes a key enzyme required for the de novo synthesis of phosphatidylinositol. Induction of INO1 resulted in acetylation of both histones H3 and H4 at the INO1 promoter and sequences farther downstream in the coding region, suggesting a gene-wide acetylation in response to transcriptional activation. Such chromatin remodeling activity requires the presence of transcriptional activator Ino2p. This indicates that histone acetylation is an activator-dependent event. Furthermore, the increase of histone acetylation is due to the increase of acetylation levels per nucleosome rather than the increase of nucleosome density. Therefore, these observations constitute evidence for the molecular mechanism of the correlation between histone acetylation and INO1 activity. PMID- 20018176 TI - Squalene-based oil-in-water emulsion adjuvants perturb metabolism of neutral lipids and enhance lipid droplet formation. AB - Oil-in-water emulsions are used as vaccine adjuvants, but the mechanism of action remains unknown. In this paper we used phagocytes (monocytes, macrophages, dendritic cells) and non-phagocytic cells (fibroblasts, skeletal muscle cells) to study internalization of emulsions in vitro, and to characterize the influence of emulsion uptake on cellular metabolism of neutral lipids. We found that all tested cell types endocytose the emulsion droplets, and that the uptake leads to an acute accumulation of neutral lipids in the form of cytoplasmic lipid droplets. The accumulated lipids comprise not only the delivered squalene, but also cholesteryl esters, triacylglycerols, fatty acids, and diacylglycerols. Lipid metabolism and innate immunity are closely linked, and accumulation of lipids in non-adipose tissues is known to induce inflammatory conditions. We propose that one aspect of o/w emulsion adjuvanticity could depend on their ability to rapidly change lipid metabolism of the target cells. PMID- 20018177 TI - Effect of simvastatin on cholesterol metabolism in C2C12 myotubes and HepG2 cells, and consequences for statin-induced myopathy. AB - The mechanism of statin-induced skeletal muscle myopathy is poorly understood. We investigated how simvastatin affects cholesterol metabolism, ubiquinone levels, and the prenylation and N-linked glycosylation of proteins in C2C12 myotubes. We used liver HepG2 cells for comparison, as their responses to statins are well characterized in terms of their cholesterol metabolism (in contrast to muscle cells), and statins are well-tolerated in the liver. Differences between the two cell lines could indicate the mechanism behind statin-induced myopathy. Simvastatin reduced de novo cholesterol production in C2C12 myotubes by 95% after 18h treatment. The reduction was 82% in the HepG2 cells. Total cholesterol pools, however, remained constant in both cell lines. Simvastatin treatment similarly did not affect total ubiquinone levels in the myotubes, unlike in HepG2 cells (22% reduction in CoQ10). Statin treatment reduced levels of Ras and Rap1 prenylation in both cell lines, whereas N-linked glycosylation was only affected in C2C12 myotubes (21% reduction in rate). From these observations, we conclude that total cholesterol and ubiquinone levels are unlikely to be involved in statin-mediated myopathy, but reductions in protein prenylation and especially N linked glycosylation may play a role. This first comparison of the responses to simvastatin between liver and skeletal muscle cell lines may be important for future research directions concerning statin-induced myopathy. PMID- 20018178 TI - The modulatory effects of bHLH transcription factors with the Wnt/beta-catenin pathway on differentiation of neural progenitor cells derived from neonatal mouse anterior subventricular zone. AB - The subventricular zone (SVZ) located adjacent to the lateral ventricles is the major site where neural progenitor cells (NPCs) are concentrated in the adult brain. NPCs in the anterior subventricular zone (SVZa) generate neuronal precursors and migrate along a highly localized pathway--the rostral migratory stream (RMS) to the olfactory bulb (OB), where they differentiate into interneurons. To investigate the modulatory effects of basic helix-loop-helix (bHLH) transcription factors on differentiation from SVZa NPCs, we firstly examined the distribution of bHLH family members (Mash1, Id2, and Hes1) in cultured mouse SVZa NPCs and evaluated their regulatory effects on differentiation by transfection with Mash1, Id2, or Hes1 eukaryotic expression plasmid. Furthermore, we assessed the effects of bHLH transcription factors on the expression of downstream molecules of the Wnt/beta-catenin pathway, beta catenin and (Glycogen synthase kinase-3beta). Our results demonstrated that Mash1, Id2, Hes1 were all widely expressed in in vitro progenies from mouse SVZa NPCs. Analyses of SVZa NPCs transfected with eukaryotic expression plasmids showed that Mash1 promoted neuronal differentiation from SVZa NPCs, while Id2 and Hes1 repressed neuronal differentiation. In addition, we found that Id2 and Hes1 simulated expression of beta-catenin and GSK-3beta, while Mash1 inhibited their expression. Our results suggest that the classic bHLH transcription factors, Mash1, Id2 and Hes1, play important roles in the regulation of differentiation from SVZa NPCs. This modulation is possibly mediated by a coordination of bHLH and Wnt/beta-catenin signaling. PMID- 20018180 TI - Age-effects on associative object-location memory. AB - Aging is accompanied by an impairment of associative memory. The medial temporal lobe and fronto-striatal network, both involved in associative memory, are known to decline functionally and structurally with age, leading to the so-called associative binding deficit and the resource deficit. Because the MTL and fronto striatal network interact, they might also be able to support each other. We therefore employed an episodic memory task probing memory for sequences of object location associations, where the demand on self-initiated processing was manipulated during encoding: either all the objects were visible simultaneously (rich environmental support) or every object became visible transiently (poor environmental support). Following the concept of resource deficit, we hypothesised that the elderly probably have difficulty using their declarative memory system when demands on self-initiated processing are high (poor environmental support). Our behavioural study showed that only the young use the rich environmental support in a systematic way, by placing the objects next to each other. With the task adapted for fMRI, we found that elderly showed stronger activity than young subjects during retrieval of environmentally richly encoded information in the basal ganglia, thalamus, left middle temporal/fusiform gyrus and right medial temporal lobe (MTL). These results indicate that rich environmental support leads to recruitment of the declarative memory system in addition to the fronto-striatal network in elderly, while the young use more posterior brain regions likely related to imagery. We propose that elderly try to solve the task by additional recruitment of stimulus-response associations, which might partly compensate their limited attentional resources. PMID- 20018179 TI - Role of gap junctions in early brain injury following subarachnoid hemorrhage. AB - Gap junction inhibition has been demonstrated to reverse the vascular contraction that follows experimental subarachnoid hemorrhage. This study hypothesizes that the use of established gap junction inhibitors: octonal and carbenoxolone, to interrupt cell to cell communication will provide neuroprotection against early brain injury after SAH. The filament perforation model of SAH was performed in male Sprague-Dawley rats weighing between 300 and 380 g. Octanol (260.46 mg or 781.38 mg/kg), carbenoxolone (100 mg/kg), or vehicles were given via intraperitoneal injection 1 h after SAH. Neurologic deficits and cerebral apoptosis were assessed 24 and 72 h after SAH. In addition, Western blot analysis was performed to confirm the in vivo inhibition of CNS gap junctions. The administration of octanol and carbenoxolone both failed to attenuate the neurological deficits induced by SAH, and they did not reduce neuronal apoptosis. Additionally, carbenoloxone increased post SAH mortality and exacerbated SAH induced apoptosis. Despite previous studies that show gap junction inhibitors reverse vasospasm following experimental SAH, they failed to improve clinical outcomes or provide neuroprotection in this study. PMID- 20018181 TI - Administration of thyroid hormone increases reelin and brain-derived neurotrophic factor expression in rat hippocampus in vivo. AB - Thyroid hormones play important roles in the maturation and function of the central nervous system. However, the underlying mechanism behind thyroid hormone regulated gene expression in the adult brain is not well understood. Two genes critical for neuronal plasticity and implicated in psychiatric disorders, reelin and brain-derived neurotrophic factor (BDNF), were investigated in the present study. Triiodothyronine (T3), the active form of thyroid hormone was administered to young adult rats in two different manners: systemic injection or local brain infusion. Real time RT-PCR results revealed that T3 administration lead to a significant increase in reelin, total BDNF and exon-specific BDNF mRNA expression in the hippocampus. Furthermore, the association of transcriptional coactivators (including steroid receptor coactivator-1 (SRC-1), cAMP response element binding protein-binding protein (CBP), and thyroid hormone receptor associated protein 220 (TRAP 220)) and RNA polymerase II (RNA Pol II), with reelin and BDNF genes in the rat hippocampus displayed a distinct process following thyroid hormone administration. These findings suggest that association of transcriptional coactivators and RNA Pol II with gene promoters may be a possible mechanism explaining T3-induced reelin and BDNF expression in the hippocampus of young adult rats. PMID- 20018182 TI - 3-(4-(Benzo[d]thiazol-2-yl)-1-phenyl-1H-pyrazol-3-yl) phenyl acetate induced Hep G2 cell apoptosis through a ROS-mediated pathway. AB - 3-(4-(Benzo[d]thiazol-2-yl)-1-phenyl-1H-pyrazol-3-yl) phenyl acetate (DPB-5) is a synthetic benzothiazole derivative. In the present study, we revealed that DPB-5 had strong cytotoxicity to induce cell apoptosis, which was mediated by ROS. And DPB-5 was more cytotoxic toward hepatoma cells than toward normal hepatic cells, which was resulted from the greater susceptibility of the malignant cells to ROS. DBP-5 caused massive ROS accumulation and GSH decrease, which lead to MMP disruption, caspase activation and finally induced cell apoptosis. Additionally, rotenone, an inhibitor of mitochondria electron transport system, effectively blocked the ROS elevated effect of DPB-5, which suggested that DPB-5-induced ROS generated from the mitochondria. Further studies showed that DPB-5-induced cell apoptosis through caspases-cascade, but failed to activate caspase-9. Hence, we concluded that DPB-5-induced Hep G2 cells apoptosis via a ROS-mediated pathway which was caspase-dependent but did not rely on caspase-9. PMID- 20018183 TI - Hematopoietic neoplastic diseases develop in C3H/He and C57BL/6 mice after benzene exposure: strain differences in bone marrow tissue responses observed using microarrays. AB - In this study, Trp53-deficient and wild-type mice of both C57BL/6 and C3H/He strains were exposed to benzene (33, 100, and 300 ppm; 6h/day, 5 days/week for 26 weeks) and then observed for lifetime. As results, first, the incidence of nonthymic lymphomas in C57BL/6 mice and acute myeloid leukemias (AMLs) in C3H/He mice showed linear responses at the lower exposure level in Trp53-deficient mice; second, the incidence of thymic lymphomas in C57BL/6 mice and nonthymic lymphomas in C3H/He mice increased without a plateau-like ceiling; thus, the former equivocal induction of hematopoietic neoplasms (HPNs) in the case of low-dose benzene exposure was assumed to be based on the DNA repair potential in wild-type mice, and the latter limited increase in HPNs in the case of high-dose benzene exposure was considered to be due to excessive apoptosis in wild-type mice. Concerning the incidence of AMLs, though a dose of 300 ppm benzene inhalation induced 9% AMLs in wild-type C3H/He mice-AML-prone, it induced AMLs in 38% of Trp53-deficient C3H/He mice. Because AMLs were also observed in Trp53-deficient mice, including in the C57BL/6 mice, benzene exposure may also be a potent inducer of AMLs in mice with some strain differences. In the present study, to elucidate the hematopoietic stem cell-specific, aryl hydrocarbon-receptor-related low-dose adverse effect, global gene expression in the bone marrow was analyzed at 28 days after 2-week-intermittent exposure to 150 mg/kg b.w. benzene, by gavage, i.e., equivalent to the above inhalation protocol with 300 ppm. We observed two conceptually different gene expression profiles; "common gene profiles" (CGPs) shared among mice in each group, and "stochastic gene profiles" (SGPs), i.e., unique union genes from one individual mouse to another. The CGPs of the experimental group and the SGPs of each individual mouse were separately characterized by individual assay. Concerning the CGPs, reciprocal strain differences between C3H/He and C57BL/6 mice in expression gene profiles, both plausible for leukemogenesis, were identified; namely, dominant downmodulations of Sltm and Cryl1, related to suppression of apoptosis and genomic instability in C3H/He mice, respectively, and dominant downmodulations of Atrx/rad54 and Kdm2a, related to a decrease in DNA repair and genomic instability, respectively, in C57BL/6 mice. These findings imply that these reciprocal gene expression differences induced by benzene exposure may lead each strain to undergo different hematopoietic neoplastic pathways. In contrast, each individual mouse often shows a unique SGP. SGPs often include transcription factors, which regulate reciprocal signaling pathways including further SGPs. Among them, apoptosis-related genes expressed in C57BL/6 mice and those in C3H/He mice were attributable to different combinations of SGPs. Such stochastic case-by-case gene expression may be in good agreement with the individual and strain differences observed following benzene exposure. Because gene chip microarray techniques can elucidate stochastic changes in gene expression profiles, possible stochastic toxicology and its future role are discussed. PMID- 20018184 TI - Quantification of phase transitions of lipid mixtures from bilayer to non-bilayer structures: Model, experimental validation and implication on membrane fusion. AB - Lipid bilayers provide a solute-proof barrier that is widely used in living systems. It has long been recognized that the structural changes of lipids during the phase transition from bilayer to non-bilayer have striking similarities with those accompanying membrane fusion processes. In spite of this resemblance, the numerous quantitative studies on pure lipid bilayers are difficult to apply to real membranes. One reason is that in living matter, instead of pure lipids, lipid mixtures are involved and there is currently no model that establishes the connection between pure lipids and lipid mixtures. Here, we make this connection by showing how to obtain (i) the short-range repulsion between bilayers made of lipid mixtures and, (ii) the pressure at which transition from bilayer phase to non-bilayer phases occur. We validated our models by fitting the experimental data of several lipid mixtures to the theoretical data calculated based on our model. These results provide a useful tool to quantitatively predict the behavior of complex membranes at low hydration. PMID- 20018185 TI - Polysialic acid can mediate membrane interactions by interacting with phospholipids. AB - Polysialic acid (polySia) is expressed on the surface of neural cells, neuroinvasive bacterial cells and several tumor cells. PolySia chains attached to NCAM can influence both trans interactions between membranes of two cells and cis interactions. Here, we report on the involvement of phospholipids in regulation of membrane interactions by polySia. The pH at the surface of liposomes, specific molecular area of phosphatidylcholine molecules, phase transition of DPPC bilayers, cyclic voltammograms of BLMs, and electron micrographs of phosphatidylcholine vesicles were studied after addition of polysialic acid free in solution. The results indicate that polySia chains can associate with phosphatidylcholine bilayers, incorporate into the polar part of a phospholipid monolayer, modulate cis interactions between phosphatidylcholine molecules, and facilitate trans interactions between apposing phospholipid vesicles. These observations imply that polySia attached to NCAM or to lipids can behave similarly. PMID- 20018186 TI - Reduced serum vaspin concentrations in obese children following short-term intensive lifestyle modification. AB - BACKGROUND: Recently, visceral adipose tissue-derived serpin (vaspin) was identified as a potential insulin sensitizing adipokine, however, the factors determining the levels of circulating vaspin levels have not been fully understood. We investigated the association between adiposity, insulin resistance, lipid profiles and inflammatory markers including vaspin levels, and the effects of short-term intensive lifestyle modification on circulating vaspin levels in overweight or obese children. METHODS: A total of 50 (25 boys, 25 girls) overweight or obese children aged 11 to 13 years (average age: 12.0+/-0.9 y, BMI: 25.35+/-86 kg/m(2)) who complied with inclusion criteria participated in our study. To determine the association between adiposity, insulin resistance, lipid profiles and inflammatory markers including vaspin levels, cross-sectional analyses were performed. Thereafter, subjects underwent a tightly controlled seven-day intensive lifestyle modification including physical activity, dietary modification, and behavioral modification education in residence of a local university dormitory. RESULTS: There was a negative correlation between vaspin concentration and fasting insulin (r=-.325, p<0.05) and homeostasis model assessment of insulin resistance (HOMA-IR) (r=-.331, p<0.05) when percent body fat was controlled. Multivariate linear regression analysis found serum vaspin level to be an independent predictor of insulin and HOMA-IR. Short-term intensive lifestyle modification significantly decreased vaspin levels by 39.28% (pre: .84+/-1.0, post: .51+/-1.0 ng/ml, p<0.001) while adiponectin levels increased by 11.2% (pre: 6.50+/-2.89, post: 7.28+/-2.98 ng/ml, p<0.01). In addition, short term lifestyle modification significantly improved HOMA-IR (pre: 3.58+/-1.93, post 1.30+/-1.9, p<0.001) and lipid profiles. CONCLUSIONS: Serum vaspin level is one of the predictors for insulin resistance and was significantly reduced following short-term lifestyle modification. PMID- 20018187 TI - The [FeFe]-hydrogenase maturase HydF from Clostridium acetobutylicum contains a CO and CN- ligated iron cofactor. AB - Biosynthesis of the [FeFe] hydrogenases active site (H-cluster) requires three maturation factors whose respective roles are not understood yet. The clostridial maturation enzymes (CaHydE, CaHydF and CaHydG) were homologously overexpressed in their native host Clostridium acetobutylicum. CaHydF was able to activate Chlamydomonas reinhardtii [FeFe] hydrogenase apoprotein (CrHydA1(apo)) to almost 100% compared to the native specific hydrogen evolution activity. Based on electron paramagnetic resonance spectroscopy and Fourier-transform infrared spectroscopy data the existence of a [4Fe4S] cluster and a CO and CN(-) ligand coordinated di-iron cluster is suggested. This study contains the first experimental evidence that the bi-nuclear part of the H-cluster is assembled in HydF. PMID- 20018189 TI - A novel kinase inhibitor establishes a predominant role for protein kinase D as a cardiac class IIa histone deacetylase kinase. AB - Class IIa histone deacetylases (HDACs) repress genes involved in pathological cardiac hypertrophy. The anti-hypertrophic action of class IIa HDACs is overcome by signals that promote their phosphorylation-dependent nuclear export. Several kinases have been shown to phosphorylate class IIa HDACs, including calcium/calmodulin-dependent protein kinase (CaMK), protein kinase D (PKD) and G protein-coupled receptor kinase (GRK). However, the identity of the kinase(s) responsible for phosphorylating class IIa HDACs during cardiac hypertrophy has remained controversial. We describe a novel and selective small molecule inhibitor of PKD, bipyridyl PKD inhibitor (BPKDi). BPKDi blocks signal-dependent phosphorylation and nuclear export of class IIa HDACs in cardiomyocytes and concomitantly suppresses hypertrophy of these cells. These studies define PKD as a principal cardiac class IIa HDAC kinase. PMID- 20018188 TI - C-mip interacts with the p85 subunit of PI3 kinase and exerts a dual effect on ERK signaling via the recruitment of Dip1 and DAP kinase. AB - In naive T cells, Lck exerts a negative control on the ERK/MAPK pathway. We show that c-mip (c-maf inducing protein) interacts with the p85 subunit of PI3 kinase and inactivates Lck, which results in Erk1/2 and p38 MAPK activation. This effect is not enough to activate AP1 given the inability of ERK to migrate into the nucleus and to transactivate its target genes. We demonstrate that c-mip interacts with Dip1 and upregulates DAPK, which blocks the nuclear translocation of ERK1/2. This dual effect of c-mip is unique and might represent a potential mechanism to prevent the development of an immune response. PMID- 20018190 TI - Cloning and differential expression of steroid 5 alpha-reductase type 1 (Srd5a1) and type 2 (Srd5a2) from the Harderian glands of hamsters. AB - In hamsters, the Harderian glands (HGs) exhibit a marked sexual dimorphism which is thought to depend on dihydrotestosterone (DHT); however, it is unclear whether hamster HGs contain one or more 5 alpha-reductases and whether these enzymes are differentially expressed in males and females. In this study, we isolated specific cDNAs for 5 alpha-reductase 1 (Srd5a1) and 5 alpha-reductase 2 (Srd5a2), determined their sequences and investigated their expression in the HG of both sexes. Isozyme 1, cloned from liver mRNA, encodes a protein of 255 amino acids (aa); isozyme 2 cDNA, isolated from the epididymis encodes a 254-aa protein. When assayed in transfected HEK-293 cells, the type 1 isozyme displayed activity over a broad pH range (6.5-8), while isozyme 2 had a pH optimum of 5.5. Both isoenzymes efficiently catalyzed the in vitro transformation of T into DHT, with apparent K(m) values of 7.1 and 1.9 micromol/L for Srd5a1 and Srd5a2, respectively. Real-time PCR analysis revealed higher mRNA levels for Srd5a1 than for Srd5a2. Expression of both isoenzymes increased slightly in HGs of castrated males and showed variations during the estrous cycle in females. Hormonal replacement with 17beta-estradiol administered to spayed females induced the up regulation of Srd5a2 mRNA levels. Altogether, our results demonstrated that both Srd5a1 and Srd5a2 are expressed in HGs without clear differences between males and females. The biochemical characteristics and relative expression of these 5 alpha-reductases support the view that both isozymes may play a relevant role in modulating androgen signaling in HG. PMID- 20018191 TI - Bursts of transposition from non-long terminal repeat retrotransposon families of the RTE clade in Schistosoma mansoni. AB - The genus Schistosoma is composed of blood flukes that infect vertebrates, from which three species are major causative agents of human schistosomiasis, a tropical disease that affects more than 200 million people. Current models of the recent evolution of Schistosoma indicate multiple events of migration and speciation from an Asian ancestral species. Transposable elements are important drivers of genome evolution and have been hypothesised to have an important role in speciation. In this work, we describe a comprehensive inventory of Schistosoma mansoni and Schistosoma japonicum retrotransposons, based on their recently published genomic data. We find a considerable difference in retrotransposon representation between the two species (22% and 13%, respectively). A large part of this difference can be attributed to higher representation of two previously described families of S. mansoni retrotransposons (SR2 and Perere-3/SR3), compared with the representation of their closest relative families in S. japonicum. A more detailed analysis suggests that these two S. mansoni families were the subject of recent bursts of transposition that were not paralleled by their S. japonicum counterparts. We hypothesise that these bursts could be a consequence of the evolutionary pressure resulting from migration of Schistosoma from Asia to Africa and their establishment in this new environment, helping both speciation and adaptation. PMID- 20018192 TI - Compounds of the upper gastrointestinal tract induce rapid and efficient excystation of Entamoeba invadens. AB - The infective stage of Entamoeba parasites is an encysted form. This stage can be readily generated in vitro, which has allowed identification of stimuli that trigger the differentiation of the parasite trophozoite stage into the cyst stage. Studies of the second differentiation event, emergence of the parasite from the cyst upon infection of a host, have been hampered by the lack of an efficient means to excyst the parasite and complete the life cycle in vitro. We have determined that a combination of exposures to water, bicarbonate and bile induces rapid excystment of Entamoeba invadens cysts. The high efficiency of this method has allowed the visualization of the dynamics of the process by electron and confocal microscopy, and should permit the analysis of stage-specific gene expression and high-throughput screening of inhibitory compounds. PMID- 20018193 TI - A diffusion process to model generalized von Bertalanffy growth patterns: fitting to real data. AB - The von Bertalanffy growth curve has been commonly used for modeling animal growth (particularly fish). Both deterministic and stochastic models exist in association with this curve, the latter allowing for the inclusion of fluctuations or disturbances that might exist in the system under consideration which are not always quantifiable or may even be unknown. This curve is mainly used for modeling the length variable whereas a generalized version, including a new parameter b > or = 1, allows for modeling both length and weight for some animal species in both isometric (b = 3) and allometric (b not = 3) situations. In this paper a stochastic model related to the generalized von Bertalanffy growth curve is proposed. This model allows to investigate the time evolution of growth variables associated both with individual behaviors and mean population behavior. Also, with the purpose of fitting the above-mentioned model to real data and so be able to forecast and analyze particular characteristics, we study the maximum likelihood estimation of the parameters of the model. In addition, and regarding the numerical problems posed by solving the likelihood equations, a strategy is developed for obtaining initial solutions for the usual numerical procedures. Such strategy is validated by means of simulated examples. Finally, an application to real data of mean weight of swordfish is presented. PMID- 20018194 TI - Modeling growth and telomere dynamics in Saccharomyces cerevisiae. AB - A general branching process is proposed to model a population of cells of the yeast Saccharomyces cerevisiae following loss of telomerase. Previously published experimental data indicate that a population of telomerase-deficient cells regain exponential growth after a period of slowing due to critical telomere shortening. The explanation for this phenomenon is that some cells engage telomerase independent pathways to maintain telomeres that allow them to become "survivors." Our model takes into account random variation in individual cell cycle times, telomere length, finite replicative lifespan of mother cells, and survivorship. We identify and estimate crucial parameters such as the probability of an individual cell becoming a survivor, and compare our model predictions to experimental data. PMID- 20018195 TI - Consequential classes of resources: Subtle global bifurcation with dramatic ecological consequences in a simple population model. AB - Numerous situations exist in which a consumer uses two different kinds of resources, one fixed, the other renewable, e.g., nesting resources and food resources. With an elementary modification of the basic Lotka-Volterra consumer resource equations, we investigate the population dynamics of a consumer dependent on two resources, one fixed, the other renewable. Emerging from this structure is a situation of alternative attractors that remain qualitatively robust over a significant range of parameter values. However, a dramatic change in basins of attraction is induced by very small changes in parameters due to a global bifurcation. Noteworthy is the fact that the qualitative nature of the alternative equilibria remains constant but the dramatic change in the basins does not arise from subtle differences in initial conditions. Rather, there is a major restructuring of the vector field such that a permanent change involving large sets of initial conditions results from very small changes in parameters. PMID- 20018196 TI - Temporal gene expression changes induced by a low concentration of benzo[a]pyrene diol epoxide in a normal human cell line. AB - (+ or -)-anti-Benzo[a]pyrene-7,8-diol-9,10-epoxide (BPDE), which causes bulky adduct DNA damage, is well-characterized as the ultimate carcinogen of benzo[a]pyrene (BaP). In this study, we have employed Affymetrix HG-U133 Plus 2.0 microarray and quantitative real-time RT-PCR methods to investigate a temporal transcriptomic response triggered by a low concentration (0.05 microM) of BPDE at 1, 10, and 22 h after exposure in normal human cells. The differential gene expression profiles at the three time points varied greatly, and generally reflected a cellular responsive process from initiation to progression and to recovery after the BPDE-caused damage. The dynamic regulation of the genes related with cell cycle progression and cell fate exhibited a tendency from inhibition to survival, which was accordant with the cell cycle arrest and cytotoxicity data induced by the low-dose BPDE exposure. In silico comparison of the genomic data revealed that BPDE and ultraviolet induced a panel of common transcriptional responses, which might be related with a series of similar molecular processes elicited by these two DNA-damaging agents. In conclusion, this whole-genome time-course study has identified a dynamically regulated transcriptional signature after low-dose BPDE exposure, which may help to understand the complex mechanisms of mutagenesis and carcinogenesis induced by BPDE. PMID- 20018198 TI - Separating coordinative and executive dysfunction in cerebellar patients during motor skill acquisition. AB - OBJECTIVE: Patients with cerebellar stroke are impaired in motor skill acquisition and cognitive/executive performance. The aim was to test whether skill acquisition in cerebellar patients is influenced by executive demands such as the intermittent exercise of a conflicting motor task. METHODS: Patients with cerebellar stroke and healthy controls were tested in two serial reaction time experiments. In Experiment 1, participants performed practice runs (always same sequence) and interference runs (new sequence for each run) in a strictly alternating fashion. In Experiment 2, participants rested between successive practice runs; the duration of rests was adapted to the duration of interference runs in the other experiment. Participants of Experiment 1 were also tested for cognitive-executive functions (Wisconsin Card Sort, Word Fluency, Trail Making, Digit Span backwards). RESULTS: (1) Patients in Experiment 1, although always slower than controls, acquired motor skills in the first run before interference but in contrast to controls failed to improve their performance in subsequent runs. (2) Patients in Experiment 2 improved their performance consistently over several runs. (3) Patients of Experiment 1 were worse than controls in several cognitive-executive functions; however, these deficits did not correlate with the degree of interference in motor skill acquisition. INTERPRETATION: Simple movement coordination and higher order context-related movement organisation are separate cerebellar functions. In cerebellar patients, impaired movement coordination is associated with generally slower reaction times whereas organisational deficits are associated with a specific impairment to change between motor sets. Motor-executive functions responsible for the latter impairment might be independent from cognitive-executive functions. PMID- 20018197 TI - From synapse to nucleus: novel targets for treating depression. AB - The need for newer compounds to treat depression is an ever-growing concern due to the enormous societal and financial ramifications of this disorder. Here, we review some of the candidate systems that could potentially be involved in depression, or an inherent resistance to depression termed resilience, and the numerous protein targets for these systems. A substantial body of literature provides strong evidence that neurotrophic factors, glutamate receptors, hypothalamic feeding peptides, nuclear hormone receptors, and epigenetic mechanisms, among others, will make for interesting targets when examining depressive behavior or resilience in preclinical models, and eventually clinical trials. Although some of these targets for depression already appear promising, new waves of more selective compounds for any molecular system should promote a better understanding of this complex disease and perhaps improved treatments. PMID- 20018199 TI - Sensorimotor function and sensorimotor tracts after hemispherectomy. AB - Hemispherectomy is currently the only effective treatment for relieving constant seizures in children with severe or progressive unilateral cortical disease. Although early hemispherectomy has been advocated to avoid general dysfunction due to continued seizures, it remains unclear whether age at surgery affects specific sensorimotor functions. Little is know about the anatomical status of sensorimotor pathways after hemispherectomy and how it might relate to sensorimotor function. Here we measured motor function and sensory thresholds of the upper and lower limbs in 12 hemispherectomized patients. Diffusion tensor imaging (DTI) was used to determine status of brainstem corticospinal tracts and medial lemniscus. Hemispherectomy subjects showed remarkable recovery in both sensory and motor function. Many patients showed normal sensory vibration thresholds. Within the smaller Rasmussen's subgroup, we saw a relationship between age at surgery and sensorimotor function recovery (i.e. earlier was better). Anatomically, we found marked asymmetry in brainstem corticospinal tracts but preserved symmetry in the medial lemniscus, which may relate to robust sensory recovery. Age at surgery predicted anatomical status of brainstem sensorimotor tracts. In sum, we found that age at surgery influences anatomical changes in brainstem motor pathways, and may also relate to sensorimotor recovery patterns. PMID- 20018200 TI - Dopaminergic medication boosts action-effect binding in Parkinson's disease. AB - Parkinson's disease (PD) is a neurodegenerative disorder affecting voluntary motor control. However, little is known about the experience of voluntary action in PD patients. A key component of action experience is the feeling of controlling one's own actions, and through them, external events. In healthy individuals this sense of agency (SoA) is associated with a subjective compression of time, such that actions and their effects are perceived as bound together across time. This action-effect binding provides an indirect measure of SoA. Nine PD patients and age-matched controls judged the time of voluntary actions and of an auditory effect (a tone) of the action. The pattern of results resembled previous studies, with the perceived time of actions showing a shift towards the subsequent tone, relative to a baseline condition involving actions without tones. Similarly, the perceived times of tones showed a shift towards the preceding action that caused the tone, relative to a baseline condition involving tones only. The patients were tested both on and off dopaminergic medication. PD patients off medication showed no significant change in action-effect binding relative to controls. Conversely, PD patients on medication showed a significant increase in action-effect binding relative to their own performance off medication. Increased availability of dopamine strengthened the experience of association between actions and external events, enhancing the sense of agency. These results shed light on the contribution of dopamine to the experience of instrumental action, and also on impulse control disorders and psychosis in medicated PD patients. PMID- 20018202 TI - Behavioural mechanisms affecting energy regulation in mice prone or resistant to diet- induced obesity. AB - We investigated inbred SWR/J and AKR/J mice, two established models for different susceptibility to diet-induced obesity (DIO), to scrutinize the contribution of physical activity and energy assimilation to the etiology of developing obesity. Body mass gain and body composition of mice fed a high-energy (HE) or a low caloric control diet were monitored. In parallel, assimilated energy, locomotor activity and thermoregulatory behaviour were measured. Activity was continuously registered by radio telemetry and, in addition, Open Field (OF) behaviour was used as a quick screening tool for spontaneous activity before and after the feeding trial. Energy assimilation was increased in both strains on HE (AKR/J: +60.7% and SWR/J: +42.8%) but only in AKR/J, body mass (+8.1%) and fat mass (+40.7%) were significantly elevated. As a trend, total home cage activity was increased and was more scattered in SWR/J. Interestingly, HE stimulated OF activity only in SWR/J in the second trial at the end of the feeding experiment. The spatial pattern of OF activity also differed between strains with obese mice avoiding the core area. Under housing conditions, nest building behaviour was more pronounced in AKR/J. To further evaluate OF behaviour as a marker for spontaneous activity an obese mouse line was investigated. Mice lacking the leptin receptor (db/db) showed already before the onset of obesity lowest activity levels in OF. Adjustment of energy intake, higher activity levels and energy consuming thermoregulatory behaviour are mechanisms employed by SWR/J mice to dissipate excess energy as a defence against the onset of obesity. Therefore our results deciphering mechanisms of DIO-sensitivity in mice contribute to the understanding of inter-individual differences in body weight development in an adipogenic environment. PMID- 20018201 TI - Callosal degradation in HIV-1 infection predicts hierarchical perception: a DTI study. AB - HIV-1 infection affects white matter circuits linking frontal, parietal, and subcortical regions that subserve visuospatial attention processes. Normal perception requires the integration of details, preferentially processed in the left hemisphere, and the global composition of an object or scene, preferentially processed in the right hemisphere. We tested whether HIV-related callosal white matter degradation contributes to disruption of selective lateralized visuospatial and attention processes. A hierarchical letter target detection paradigm was devised, where large (global) letters were composed of small (local) letters. Participants were required to identify target letters among distractors presented at global, local, both or neither level. Attention was directed to one (global or local) or both levels. Participants were 21 HIV-1 infected and 19 healthy control men and women who also underwent Diffusion Tensor Imaging (DTI). HIV-1 participants showed impaired hierarchical perception owing to abnormally enhanced global facilitation effects but no impairment in attentional control on local-global feature selection. DTI metrics revealed poorer fiber integrity of the corpus callosum in HIV-1 than controls that was more pronounced in posterior than anterior regions. Analysis revealed a double dissociation of anterior and posterior callosal compromise in HIV-1 infection: compromise in anterior but not posterior callosal fiber integrity predicted response conflict elicited by global targets, whereas compromise in posterior but not anterior callosal fiber integrity predicted response facilitation elicited by global targets. We conclude that component processes of visuospatial perception are compromised in HIV-1 infection attributable, at least in part, to degraded callosal microstructural integrity relevant for local-global feature integration. PMID- 20018203 TI - Extra-nuclear signaling of ERalpha to the actin cytoskeleton in the central nervous system. AB - Cell morphology is controlled by a complex and redundant array of intracellular signaling pathways devoted to the regulation of the actin cytoskeleton and of its relationship with the cell membrane and the extracellular matrix. Sex steroids are effective regulators of cell morphology and tissue organization, and recent evidence indicates that this is obtained through the regulation of the cytoskeleton. Intriguingly, many of these regulatory actions related to cell morphology are achieved through rapid, non-classical signaling of sex steroid receptors to kinase cascades, independently from nuclear alteration of gene expression or protein synthesis. The identification of the mechanistic basis for these rapid actions on cell cytoskeleton has special relevance for the characterization of the effects of sex steroids in physiological conditions, such as their role in the control of brain cell remodeling. Brain cell morphology is controlled by estrogens that regulate the development of neuron/neuron interconnections and dendritic spine density. This is thought to be critical for gender-specific differences in brain function and dysfunction. The recent advancements in the characterization of the molecular basis of the extra-nuclear signaling of estrogen helps to understand the role of estrogen in the brain, and may in the future turn out to be of relevance for clinical purposes. This review highlights the regulatory effects on the cytoskeleton and cell morphology of estrogens as well as the recent advances in the characterization of these mechanisms, providing insights and working hypotheses on possible clinical applications for the modulation of these pathways in the central nervous system. PMID- 20018204 TI - Assessment of herbal medicinal products: challenges, and opportunities to increase the knowledge base for safety assessment. AB - Although herbal medicinal products (HMP) have been perceived by the public as relatively low risk, there has been more recognition of the potential risks associated with this type of product as the use of HMPs increases. Potential harm can occur via inherent toxicity of herbs, as well as from contamination, adulteration, plant misidentification, and interactions with other herbal products or pharmaceutical drugs. Regulatory safety assessment for HMPs relies on both the assessment of cases of adverse reactions and the review of published toxicity information. However, the conduct of such an integrated investigation has many challenges in terms of the quantity and quality of information. Adverse reactions are under-reported, product quality may be less than ideal, herbs have a complex composition and there is lack of information on the toxicity of medicinal herbs or their constituents. Nevertheless, opportunities exist to capitalise on newer information to increase the current body of scientific evidence. Novel sources of information are reviewed, such as the use of poison control data to augment adverse reaction information from national pharmacovigilance databases, and the use of more recent toxicological assessment techniques such as predictive toxicology and omics. The integration of all available information can reduce the uncertainty in decision making with respect to herbal medicinal products. The example of Aristolochia and aristolochic acids is used to highlight the challenges related to safety assessment, and the opportunities that exist to more accurately elucidate the toxicity of herbal medicines. PMID- 20018205 TI - New directions in conventional methods of protein crystallization. AB - Novel strategies and techniques that are based on conventional crystallization methods for crystallizing proteins are described and discussed. New directions for rendering proteins and protein complexes to become more amenable to crystallization are also presented. PMID- 20018206 TI - Counterdiffusion methods applied to protein crystallization. AB - Accumulated experience during the last years on counterdiffusion crystallization methods shows that they are a convenient and generally applicable way of optimizing solution crystal growth experiments. Irrespective of whether the objective of the experiment is to improve crystal quality or size, many experiments reporting a positive or neutral effect of counterdiffusion exists, but adverse effects are consistently absent. Thus counterdiffusion is viewed as a rational crystallization approach to minimize supersaturation and impurity levels at the crystal growth front and to ensure steadiness of both values. This control of the phase transition state is automatically achieved and sustained by a dynamic equilibrium between mass transport and aggregation kinetics. The course of this function can be implemented in any media permitting diffusive mass transport (gels, capillaries, microfluidic devices or microgravity). The counterdiffusion technique has been exploited in many recent applications revealing interesting effects on nucleation and polymorphic precipitation, hence opening further possibilities for innovative screening of crystallization conditions. PMID- 20018207 TI - Mismatch-induced lethality due to a defect in Escherichia coli RecQ helicase in exonuclease-deficient background: Dependence on MutS and UvrD functions. AB - Escherichia coli DNA-unwinding protein RecQ has roles in the regulation of general recombination and the processing of stalled replication forks. In this study, we found that knockout of the recQ gene in combination with xonA xseA recJ mutations, which inhibit methyl-directed mismatch repair (MMR), caused about 100 fold increase in sensitivity to a purine analog 2-aminopurine (2AP). Intriguingly, inactivation of a MMR initiator due to the either mutation mutS or uvrD completely suppressed the 2AP sensitivity caused by recQ xonA xseA recJ mutations, suggesting that RecQ helicase might act on the DNA structures that are generated by the processing of DNA by the MutSLH complex and UvrD helicase. Moreover, the recQ gene knockout in combination with xonA xseA recJ mutations enhanced 2AP-induced filament formation, and increased by twofold the rate of spontaneous forward mutations in the thyA locus but did not increase the rate of rifampicin-resistant mutations. We discuss about the possible interplay between E. coli RecQ helicase and mismatch recognition factors. PMID- 20018209 TI - The key roles of elongases and desaturases in mammalian fatty acid metabolism: Insights from transgenic mice. AB - In mammalian cells, elongases and desaturases play critical roles in regulating the length and degree of unsaturation of fatty acids and thereby their functions and metabolic fates. In the past decade, a great deal has been learnt about these enzymes and the first part of this review summarizes our current knowledge concerning these enzymes. More recently, several transgenic mouse models lacking either an elongase (Elovl3(-/-), Elovl4(-/-), Elovl5(-/-), Elovl6(-/-)) or a desaturase (Scd-1(-/-), Scd-2(-/-), Fads2(-/-)) have been developed and the second part of this review focuses on the insights gained from studies with these mice, as well as from investigations on cell cultures. PMID- 20018208 TI - Comparative genomics of pAKD4, the prototype IncP-1delta plasmid with a complete backbone. AB - Plasmids of the incompatibility group IncP-1 are important agents of horizontal gene transfer and contribute to the spread of antibiotic resistance and xenobiotic degradation within bacterial communities. Even though some prototype plasmids have been studied in much detail, the diversity of this plasmid group was still greatly underestimated until recently, as only two of the five currently known divergent sub-groups had been described. To further improve our insight into the diversity and evolutionary history of this family of broad-host range plasmids, we compared the complete nucleotide sequence of a new IncP-1delta plasmid pAKD4 to the genomes of other IncP-1 plasmids. Plasmid pAKD4 was previously isolated by exogenous plasmid isolation from an agricultural soil in Norway. Its 56,803bp nucleotide sequence shows high similarity in gene sequence and gene order to both plasmids pEST4011 and pIJB1, the only other IncP-1delta plasmids sequenced so far. While all three plasmids have a typical IncP-1 backbone comprising replication, transfer, and stable inheritance/control genes, the low sequence similarity in some regions and presence/absence of some backbone genes compared to other IncP-1 plasmids cluster them in a divergent sub-group. Therefore this study validates the presence of a real IncP-1delta clade with multiple plasmids. Moreover, since both pEST4011 and pIJB1 are missing a portion of their transfer genes, pAKD4 represents the first completely sequenced self transferable plasmid with a complete IncP-1delta backbone. We therefore propose it to be the prototype IncP-1delta plasmid. PMID- 20018210 TI - Neuron-glia synapses in the brain. AB - The ability to investigate the electrophysiological properties of individual cells in acute brain tissue led to the discovery that many glial cells have the capacity to respond rapidly to neuronal activity. In particular, a distinct class of neuroglial cells known as NG2 cells, which exhibit many of the properties that have been described for glial subtypes such as complex cells, polydendrocytes, synantocytes and GluR cells, express ionotropic receptors for glutamate and GABA. In both gray and white matter, NG2 cells form direct synaptic junctions with axons, which enable transient activation of these receptors. Electrophysiological analyses have shown that these neuron-glia synapses exhibit all the hallmarks of 'classical' neuron-neuron synapses, including rapid activation, quantized responses, facilitation and depression, and presynaptic inhibition. Electron microscopy indicates that axons form morphologically distinct junctions at discrete sites along processes of NG2 cells, suggesting that NG2 cells are an overt target of axonal projections. AMPA receptors expressed by NG2 cells exhibit varying degrees of Ca(2+) permeability, depending on the brain region and stage of development, and in white matter NG2 cells have also been shown to express functional NMDA receptors. Ca(2+) influx through AMPA receptors following repetitive stimulation can trigger long term potentiation of synaptic currents in NG2 cells. The expression of receptors with significant Ca(2+) permeability may increase the susceptibility of NG2 cells to excitotoxic injury. Future studies using transgenic mice in which expression of receptors can be manipulated selectively in NG2 cells have to define the functions of this enigmatic neuron glia signaling in the normal and diseased CNS. PMID- 20018211 TI - Development of a blocking ELISA using a recombinant glycoprotein for the detection of antibodies to vesicular stomatitis New Jersey virus. AB - A recombinant glycoprotein (R-GP) of vesicular stomatitis New Jersey virus (VSV NJ) was expressed in insect cells by a baculovirus system. Its utility as a diagnostic antigen in a blocking ELISA was investigated as an alternative to the current native GP extracted from VSV-NJ. With the cut-off value of 73% inhibition, the R-GP ELISA exhibited 99.1% specificity for naive sera from cattle and horses. It did not cross-react with VSV-Indiana (VSV-IN) positive sera and differentiated from foot-and-mouth disease and swine vesicular disease. Taken together, this is the first report that the R-GP has a potential to be used as a diagnostic antigen in place of the native GP for the detection of antibodies to VSV-NJ in cattle and horses. PMID- 20018212 TI - Changes in tactile sensitivity over the time-course of a goal-directed movement. AB - We report three experiments designed to investigate changes in tactile sensitivity over the time-course of goal-directed movements. A dual-task paradigm involving a speeded movement task and a non-speeded perceptual task was utilized. In the movement task, participants grasped a start computer mouse with their right hand (RH) and, at the go signal, reached for and grasped a goal mouse placed 25 cm in front of it. In the perceptual task, a tactile (standard) pulse was presented to the middle finger of the left hand (LH) which was kept at rest throughout the experiment. A comparison pulse was delivered to the middle finger of the RH. In Experiment 1, this was delivered in the motor preparation period, at the release of the start mouse, during the reaching phase, at the grasp of the goal mouse, or shortly after the grasping action. In Experiment 2, the comparison pulse was delivered in the preparation period, in the early, mid or late execution periods, or in the post-movement period. In Experiment 3, participants only performed the perceptual task. The participants made an intensity comparison regarding the second pulse (i.e., stronger vs. weaker than the first). Significant changes in tactile sensitivity were observed, with decreased thresholds (i.e., better performance) during the motor preparation and post movement periods and increased thresholds (i.e., poorer performance) during the execution period. These results are discussed in terms of sensory suppression. PMID- 20018214 TI - Functional genetic analysis of the Plasmodium falciparum deoxyxylulose 5 phosphate reductoisomerase gene. AB - Novel antimalarial drugs are urgently needed to treat severe malaria caused by Plasmodium falciparum. Isoprenoid biosynthesis is a promising target pathway, since the biosynthetic route in Plasmodia is biochemically distinct from the mevalonate pathway in humans. The small molecule fosmidomycin is an inhibitor of the enzyme responsible for the first dedicated step in isoprenoid biosynthesis, deoxyxylulose 5-phosphate reductoisomerase (DXR). However, the antimalarial effects of fosmidomycin might not be specific to DXR inhibition and further validation of DXR is warranted. We present the first functional genetic validation of P. falciparum DXR (PF14_0641). Using a single cross-over strategy, we show that plasmid integration occurs at the DXR locus but only when DXR gene function is preserved, but not when integration disrupts gene function. These data indicate that DXR is required for intraerythrocytic development of P. falciparum. PMID- 20018215 TI - Mapping of regional brain activation in response to fatigue-load and recovery in rats with c-Fos immunohistochemistry. AB - Fatigue is known to be accompanied by a feeling of extreme physical or mental tiredness, resulting from severe stress and hard physical or mental work. To investigate the functional localization of neural activity related to fatigue and recovery, we examined brain c-Fos expression patterns in a rat in a state of fatigue in which rats kept in a cage filled with water to a height of 2.2cm for 1 5 days. A significant increase in the number of c-Fos-immunopositive cells was observed in the retrosplenial granular b cortex during the fatigue-loading and in the dentate gyrus of the ventral hippocampus after a 24-h recovery. In addition, variable increases in the number of c-Fos-immunopositive cells were observed in the cingulate cortex area 2, ventral part of the lateral septum nucleus, median preoptic nucleus, anterior part of the paraventricular thalamic nucleus, medial parvicellular part of the paraventricular hypothalamic nucleus, and lateral and ventrolateral periaqueductal gray during the fatigue-load period. These results indicate that such regional brain activity would be involved in fatigue or in subsequent recovery and might provide a foothold for further research into the nature of fatigue. PMID- 20018213 TI - Cellular dynamical mechanisms for encoding the time and place of events along spatiotemporal trajectories in episodic memory. AB - Understanding the mechanisms of episodic memory requires linking behavioral data and lesion effects to data on the dynamics of cellular membrane potentials and population interactions within brain regions. Linking behavior to specific membrane channels and neurochemicals has implications for therapeutic applications. Lesions of the hippocampus, entorhinal cortex and subcortical nuclei impair episodic memory function in humans and animals, and unit recording data from these regions in behaving animals indicate episodic memory processes. Intracellular recording in these regions demonstrates specific cellular properties including resonance, membrane potential oscillations and bistable persistent spiking that could underlie the encoding and retrieval of episodic trajectories. A model presented here shows how intrinsic dynamical properties of neurons could mediate the encoding of episodic memories as complex spatiotemporal trajectories. The dynamics of neurons allow encoding and retrieval of unique episodic trajectories in multiple continuous dimensions including temporal intervals, personal location, the spatial coordinates and sensory features of perceived objects and generated actions, and associations between these elements. The model also addresses how cellular dynamics could underlie unit firing data suggesting mechanisms for coding continuous dimensions of space, time, sensation and action. PMID- 20018216 TI - A mixture of peptides and sugars derived from plant cell walls increases plant defense responses to stress and attenuates ageing-associated molecular changes in cultured skin cells. AB - Small peptides and aminoacid derivatives have been extensively studied for their effect of inducing plant defense responses, and thus increasing plant tolerance to a wide range of abiotic stresses. Similarly to plants, these compounds can activate different signaling pathways in mammalian skin cells as well, leading to the up-regulation of anti-aging specific genes. This suggests the existence of analogous defense response mechanisms, well conserved both in plants and animal cells. In this article, we describe the preparation of a new mixture of peptides and sugars derived from the chemical and enzymatic digestion of plant cell wall glycoproteins. We investigate the multiple roles of this product as potential "biostimulator" to protect plants from abiotic stresses, and also as potential cosmeceutical. In particular, the molecular effects of the peptide/sugar mixture of inducing plant defense responsive genes and protecting cultured skin cells from oxidative burst damages were deeply evaluated. PMID- 20018217 TI - Assessment of siRNA pharmacokinetics using ELISA-based quantification. AB - Here, we developed a novel ELISA-based assay for quantifying double-stranded intact siRNAs for in vivo pharmacokinetic analysis. The assay makes use of dual labeled unmethylated or methylated siRNA, 5'-end-labeled on one strand with biotin (capture marker), and with dinitrophenol (detection marker), on the other end. This ELISA-based assay was linear over the range of 10-100 fmol/ml, with a sensitivity (5.4 fmol/ml) 629-fold higher than fluorometric quantification methods. The coefficient of variation (CV) of the ELISA quantification was 9.4% for intra-assay and 12.1% for inter-assay. The assay was specific for double stranded siRNAs. The intensity of the detected signal was reduced to background levels in the presence of single-stranded RNA. The ELISA-based assay revealed that the levels of methylated forms of siRNAs after transfection into A549 and HeLa cells were significantly higher than those of unmethylated siRNA forms. Applying this assay to a study of the pharmacokinetic profiles of intravenously administered siRNAs, we found that the higher blood concentrations were achieved using the methylated form of siRNAs than unmethylated form. Moreover, methylated siRNAs complexed to DOTAP-based cationic liposomes showed significantly higher and prolonged blood concentration-time profile, with 2.2-fold lower clearance rate (0.11+/-0.02 ml/min) as compared to the uncomplexed form. These results demonstrate the utility of an ELISA-based assay for evaluating chemically modified siRNAs and cationic delivery systems, particularly from a pharmacokinetic perspective. PMID- 20018219 TI - Differential expression of new splice variants of the neurotensin receptor 1 gene in human prostate cancer cell lines. AB - Neurotensin is a neuroendocrine peptide acting as a trophic factor in a variety of cells in vivo but it can also function as an autocrine growth factor in human prostate cancer cells in vitro. In addition, the high-affinity G protein-coupled NT receptor (NTS1) is overexpressed in prostate cancer cell lines. Increasing evidence argues for a direct correlation between specific alternative splice variants and cancer. We detected four splice variants of the NTS1 receptor in human prostate cancer cell lines. These isoforms include one or more exons skipping as well as an alternative 5' splice donor site and are expressed in the late-stage androgen independent prostate cancer cell lines PC3 and DU145, but not in the early-stage androgen-sensitive LNCaP or in normal prostate tissue, which only express the normal transcript. This result shows new splice variants of NTS1 for the first time. The differential expression observed among prostate cancer cell lines and normal prostate tissue opens the interesting possibility of a new role of NT/NTS1 pathway in prostate cancer. PMID- 20018218 TI - Inhibition of CD26/DPP IV attenuates ischemia/reperfusion injury in orthotopic mouse lung transplants: the pivotal role of vasoactive intestinal peptide. AB - The T cell activation Ag CD26/dipeptidylpeptidase IV (DPP IV) combines co stimulatory and enzymatic properties. Catalytically, it functions as an exopeptidase, modulating biological activity of key chemokines and peptides. Here we investigated the effect of organ-specific inhibition of DPP IV catalytic activity on ischemia/reperfusion injury after extended ischemia in the mouse model of orthotopic single lung transplantation. C57BL/6 mice were syngeneically, transplanted, grafts were perfused and stored in Perfadex with (treated) or without (control) a DPP IV enzymatic activity inhibitor (AB192). Transplantation was performed after 18h cold ischemia time; following 2-h reperfusion, grafts were analyzed for oxygenation, thiobarbituric acid-reactive substances, histomorphology, and immunohistochemistry was performed for leukocyte Ag 6, myeloperoxidase, hemoxygenase 1, vasoactive intestinal protein (VIP), and real time PCR for VIP. Treatment with the DPP IV inhibitor AB192 resulted in significant improvement of gas exchange, less lipid oxidation, preservation of parenchymal ultrastructure, reduced neutrophil infiltration, reduced myeloperoxidase expression, increased hemoxygenase 1 expression, pronounced expression of VIP in alveolar macrophages and increased mRNA expression of VIP. Inhibition of intragraft DPP IV catalytic activity with AB192 strikingly ameliorates ischemia/reperfusion injury after extended ischemia. Furthermore, preservation of endogenous intragraft VIP levels correlate with maintaining lung function and structural integrity. PMID- 20018220 TI - Antidepressant-like effects of 3,6'-disinapoyl sucrose on hippocampal neuronal plasticity and neurotrophic signal pathway in chronically mild stressed rats. AB - Recent studies suggest that the behavioral effects of chronic antidepressant treatment are mediated by stimulation of hippocampal neuronal plasticity and neurogenesis. The present study was designed to examine the effects of 3,6' disinapoyl sucrose (DISS), a bioactive component of Polygala tenuifolia Willd, on the expressions of four plasticity-associated genes: cell adhesion molecule L1 (CAM-L1), laminin, cAMP response element binding protein (CREB) and brain-derived neurotrophic factor (BDNF) in hippocampus, all of which are involved in neuronal plasticity and neurite outgrowth. We confirmed that chronic stress in rats caused a reduction in sensitivity to reward (sucrose consumption) and a decrease in mRNA levels of CAM-L1, laminin, and BDNF, together with a decrease in protein levels of phosphorylated CREB and BDNF. Repeated administration of DISS for 21 days at doses of 5, 10 and 20mg/kg reversed stress-induced alterations in sucrose consumption and these target mRNA and protein levels. In conclusion, increased expressions in the hippocampus of three noradrenergic-regulated plasticity genes and one neurotrophic factor may be one of the molecular and cellular mechanisms underlying the antidepressant action of DISS in chronic mild stress (CMS) rats. PMID- 20018221 TI - Homocysteine induces X-box-binding protein 1 splicing in the mice brain. AB - Increasing evidence has been suggested that hyperhomocysteinemia is a risk factor of neurodegenerative diseases, although, the underlying mechanisms have not been elucidated. Here, we found peripheral application of homocysteine increases X-box binding protein 1 (XBP1) splicing in the several areas of the mice brain, such as hippocampus, hypothalamus and cortex. Time-course experiments indicated that XBP1 splicing was observed from 2h, which was decreased thereafter. On the other hand, we did not observe GRP78 or CHOP induction in homocysteine-treated mice brain. As XBP1 is spliced in response to endoplasmic reticulum (ER) stress and ER stress has been implicated in the pathogenesis of CNS diseases such as Alzheimer's disease and Parkinson's disease, homocysteine-induced XBP1 splicing would be a key mechanism for such diseases. PMID- 20018222 TI - Association of polymorphism in FcGR3A gene and progression of low-grade precursor lesions of cervical carcinoma. AB - Polymorphisms in receptors of the constant part of antibodies (FcR) have been associated with susceptibility to disease and viral infections but have not been studied in cervical carcinogenesis. The distribution of the polymorphism V158F (rs396991) in FcGR3A in cervical smears was detected in a group of 84 women with stable or regressed low-grade squamous intraepithelial lesions (group I) and a group of 54 women with high-grade squamous intraepithelial lesions (HSIL) (group II). Human papillomavirus (HPV) genotyping was also performed. In 27.4% of women from group I, FF genotype was found, whereas this genotype was observed in 51.9% of patients in group II (p = 0.003; odds ratio = 2.856 (95% confidence interval = 1.4-5.8)). When only women infected with high-risk HPV were analyzed these differences were found to be even higher (p = 0.0013; odds ratio = 3.8 (95% confidence interval = 1.7-8.8)). FF genotype in FcGR3A gene seems to be associated with increased risk of low-grade squamous intraepithelial lesions to HSIL progression suggesting that its presence may play a role in HPV tolerance, persistent infection, and HSIL development. PMID- 20018223 TI - Further considerations on the evaluation of potential reduced-risk tobacco products. Part II: Re-assessment of a heuristic using the CPS-II database. AB - In a previous analysis (see Part I) we proposed a heuristic for assessing the efficacy of potential reduced-risk tobacco products (PRRPs) on lung cancer (LC) rates, using smoking cessation data published in a report from the Iowa Women's Health Study (IWHS) as a basis for sample size estimates. In this study, an additional analysis was performed using cessation data from the much larger Cancer Prevention Study II (CPS-II), which also provides data on different durations of cessation. Statistical methods were used to assess whether smokers switching to a PRRP would reduce their risk of LC. Furthermore, non-inferiority tests compared the LC risk in switchers to that in smokers who had quit smoking. The present work shows that similar sample size estimates were obtained whether the analysis was based on the IWHS or the CPS-II data sets, suggesting that the heuristic may be generally applicable to prospective real-life studies to evaluate PRRPs. Non-inferiority testing of switchers compared with quitters required approximately 10-fold more subjects than did superiority testing of switchers compared with smokers. Altogether, these estimates indicate that it is feasible, in terms of study duration and sample size, to clinically assess the LC risk-reducing potential of a PRRP. PMID- 20018224 TI - Hypotheses and fundamental study design characteristics for evaluating potential reduced-risk tobacco products. Part I: Heuristic. AB - The risk-reducing effect of a potential reduced-risk tobacco product (PRRP) can be investigated conceptually in a long-term, prospective study of disease risks among cigarette smokers who switch to a PRRP and in appropriate comparison groups. Our objective was to provide guidance for establishing the fundamental design characteristics of a study intended to (1) determine if switching to a PRRP reduces the risk of lung cancer (LC) compared with continued cigarette smoking, and (2) compare, using a non-inferiority approach, the reduction in LC risk among smokers who switched to a PRRP to the reduction in risk among smokers who quit smoking entirely. Using standard statistical methods applied to published data on LC incidence after smoking cessation, we show that the sample size and duration required for a study designed to evaluate the potential for LC risk reduction for an already marketed PRRP, compared with continued smoking, varies depending on the LC risk-reducing effectiveness of the PRRP, from a 5-year study with 8000-30,000 subjects to a 15-year study with <5000 to 10,000 subjects. To assess non-inferiority to quitting, the required sample size tends to be about 10 times greater, again depending on the effectiveness of the PRRP. PMID- 20018225 TI - Prospects for epigenetic research within cohort studies of psychological disorder: a pilot investigation of a peripheral cell marker of epigenetic risk for depression. AB - Cohort studies have considerable prima facie value for investigating epigenetic processes in psychological disorder; however, the future prospects for such studies will depend on valid peripheral markers. The purpose of this pilot study was to investigate association between buccal cell methylation and risk for depression. Epigenotyping was limited to promoter methylation of the serotonin transporter gene (5HTT). A transcription limiting VNTR in the 5HTT promoter (5HTTLPR) was also genotyped. A nested sample of 25 depressed and 125 non depressed adolescents was drawn from an established longitudinal study of adolescent health. There was no association between depressive symptoms and either buccal cell 5HTT methylation or 5HTTLPR. However, depressive symptoms were more common among those with elevated buccal cell 5HTT methylation who carried 5HTTLPR short-allele (OR 4.9, CI 1.9-13, p=0.001). Both complete and partial (as little as 10%) methylation of a 5HTT reporter gene in an expressing cell line reduced 5HTT activity. Replication is needed. PMID- 20018226 TI - Emotion and the auditory brainstem response to speech. AB - Effects of emotion have been reported as early as 20 ms after an auditory stimulus onset for negative valence, and bivalent effects between 30 and 130 ms. To understand how emotional state influences the listener's brainstem evoked responses to speech, subjects looked at emotion-evoking pictures while listening to an unchanging auditory stimulus (danny). The pictures (positive, negative, or neutral valence) were selected from the IAPS database and controlled for dominance and arousal. Utilizing an array of measurements to assess subcortical modulation, we have found that emotion does not substantially alter brainstem alter although there is a subtle effect of background noise suppression in both emotional conditions. PMID- 20018227 TI - Characterization of spinal afferent neurons projecting to different chambers of the rat heart. AB - The pattern of distribution of spinal afferent neurons (among dorsal root ganglia DRGs) that project to anatomically and functionally different chambers of the rat heart, as well as their morphological and neurochemical characteristics were investigated. Retrograde tracing using a patch loaded with Fast blue (FB) was applied to all four chambers of the rat heart and labeled cardiac spinal afferents were characterized by using three neurochemical markers. The majority of cardiac projecting neurons were found from T1 to T4 DRGs, whereas the peak was at T2 DRG. There was no difference in the total number of FB-labeled neurons located in ipsilateral and contralateral DRGs regardless of the chambers marked with the patch. However, significantly more FB-labeled neurons projected to the ventricles compared to the atria (859 vs. 715). The proportion of isolectin B(4) binding in FB-labeled neurons was equal among all neurons projecting to different heart chambers (2.4%). Neurofilament 200 positivity was found in greater proportions in DRG neurons projecting to the left side of the heart, whereas calretinin-immunoreactivity was mostly represented in neurons projecting to the left atrium. Spinal afferent neurons projecting to different chambers of the rat heart exhibit a variety of neurochemical phenotypes depending on binding capacity for isolectin B(4) and immunoreactivity for neurofilament 200 and calretinin, and thus represent important baseline data for future studies. PMID- 20018228 TI - Behavioral and neurobiological studies on the male progeny of maternal rats exposed to chronic unpredictable stress before pregnancy. AB - Studies have shown that maternal chronic stress or depression is linked to an increased risk for affective disorders in progeny. However, the impact of maternal chronic stress before pregnancy on their progeny in animal models has not been well studied. We investigated the behaviors and the neurobiology in 60 day-old male progeny of maternal rats exposed to a 21-day chronic unpredictable stress (CUS) before pregnancy, with male progeny of unstressed maternal rats as the control. Sucrose consumption test showed that both sucrose intake and sucrose consumption percentage of the CUS progeny were lower than those of the control progeny (P<0.05). The number of times crossing the removed hidden platform in the CUS progeny was significantly fewer than that in the control progeny in Morris water maze test (P<0.05). The level of 5-hydroxytryptamine (5-HT) in the hypothalamus was reduced but the level of norepinephrine (NE) in the hippocampus was increased in CUS progeny when compared to the control (P<0.05). Western blotting showed that the relative level of phosphorylated CREB (P-CREB) in the CUS progeny was lower than that in the control progeny (P<0.05). There were significant positive correlations between sucrose consumption percentage and the level of 5-HT in hypothalamus P<0.05) or the level of P-CREB in hippocampus (P<0.05). In conclusion, depression or stressful events before pregnancy was also associated with high risk of depression in progeny, and the down-regulation of P CREB in the hippocampus might be one of the mechanisms underlying depression in the CUS progeny. PMID- 20018229 TI - Oxytocin response to an experimental psychosocial challenge in adults exposed to traumatic experiences during childhood or adolescence. AB - Long-term implications of the exposure to traumatizing experiences during childhood or adolescence, such as sexual abuse, or cancer, have been documented, namely the subjects' response to an acute stress in adulthood. Several indicators of the stress response have been considered (e.g. cortisol, heart rate). Oxytocin (OT) response to an acute stress of individuals exposed to trauma has not been documented. Eighty subjects (n=26 women who had experienced episodes of child abuse, n=25 men and women healthy survivors of cancer in childhood or adolescence, and 29 controls) have been submitted to a laboratory session involving an experimental stress challenge, the Trier social stress test. Overall, there was a clear OT response to the psychosocial challenge. Subjects having experienced a childhood/adolescence life-threatening illness had higher mean levels of OT than both abused and control subjects. There was a moderate negative relationship between OT and salivary cortisol. It is suggested that an acute stress stimulates OT secretion, and that the exposure to enduring life threatening experiences in childhood/adolescence has long-lasting consequences regarding the stress system and connected functions, namely the activation of OT secretion. Better knowledge of such long-term implications is important so that to prevent dysregulations of the stress responses, which have been shown to be associated to the individual's mental health. PMID- 20018230 TI - Differential NR2B subunit expression at dorsal root and ventrolateral funiculus synapses on lumbar motoneurons of neonatal rat. AB - Synapse specific differences in NR2 subunit expression exist in several systems within the mammalian CNS. Here we have studied such differences on motoneurons in the neonatal rat cord using ifenprodil known to inhibit voltage-, use- and glycine-independent responses mediated by NR2B-containing N-methyl-d-aspartate receptors (NMDARs) with high specificity. In neonatal rats (P1-P9), the synapses made by the dorsal root (DR) fibres were more sensitive to ifenprodil than ventrolateral funiculus (VLF) connections on the same motoneuron. DR connections exhibited very little additional blockade to bath-applied MK-801 whereas VLF connections displayed a further decrease in amplitude. This suggests that at this immediate postnatal age, DR synapses on motoneurons contain a higher proportion of ifenprodil-sensitive diheteromeric NR1/NR2B receptors than VLF synapses. Since DR synapses have been shown in other studies to be less mature than VLF synapses on the same motoneuron at this developmental stage, these data are interpreted as indicating that less mature NMDA receptors feature a higher proportion of NR2B subunits which declines as the synapse matures. This novel finding of staggered development of NMDA receptors from different synaptic inputs on the same motoneuron is discussed in the context of its developmental and functional implications. PMID- 20018231 TI - WldS can delay Wallerian degeneration in mice when interaction with valosin containing protein is weakened. AB - Axon degeneration is an early event in many neurodegenerative disorders. In some, the mechanism is related to injury-induced Wallerian degeneration, a proactive death program that can be strongly delayed by the neuroprotective slow Wallerian degeneration protein (Wld(S)) protein. Thus, it is important to understand the Wallerian degeneration mechanism and how Wld(S) blocks it. Wld(S) location is influenced by binding to valosin-containing protein (VCP), an essential protein for many cellular processes including membrane fusion and endoplasmic reticulum associated degradation. In mice, the N-terminal 16 amino acids (N16), which mediate VCP binding, are essential for Wld(S) to protect axons, a role which another VCP binding sequence can substitute. In Drosophila, the Wld(S) phenotype is weakened by a similar N-terminal truncation and by knocking down the VCP homologue ter94. Neither null nor floxed VCP mice are viable so it is difficult to confirm the requirement for VCP binding in mammals in vivo. However, the hypothesis can be tested further by introducing a Wld(S) missense mutation, altering its affinity for VCP but minimizing the risk of disturbing other aspects of its structure or function. We introduced the R10A mutation, which weakens VCP binding in vitro, and expressed it in transgenic mice. R10AWld(S) fails to co immunoprecipitate VCP from mouse brain, and only occasionally and faintly accumulates in nuclear foci for which VCP binding is necessary but not sufficient. Surprisingly however, axon protection remains robust and indistinguishable from that in spontaneous Wld(S) mice. We suggest that either N16 has an additional, VCP-independent function in mammals, or that the phenotype requires only weak VCP binding which may be driven forwards in vivo by the high VCP concentration. PMID- 20018232 TI - Pilocarpine-induced status epilepticus causes acute interneuron loss and hyper excitatory propagation in rat insular cortex. AB - Recent clinical studies have shown that the insular cortex (IC) is involved in temporal lobe epilepsy and suggested that the IC mediates spreading of epileptic activity from the temporal lobe, including the hippocampus and amygdala, to the frontal cortex. However, little is known about anatomical and physiological features of the IC in models of temporal lobe epilepsy. The present study evaluated the distribution pattern of GABAergic interneurons, especially parvalbumin (PV)- and somatostatin (SS)-immunopositive neurons, and excitatory propagation pattern in the IC of rats 4-7 days and 2 months after pilocarpine induced status epilepticus (4-7 d and 2 m post-SE rats, respectively). The number of PV-immunopositive neuron profiles in the agranular IC (AI) significantly decreased by 24.6% and 41.5% in 7 d and 2 m post-SE rats, respectively. The dysgranular and granular IC (DI+GI) exhibited only 5.2% loss of PV-immunopositive neurons in 7 d post-SE rats, while 2 m post-SE rats showed 30.4% loss of PV immunopositive neurons. There was no significant change of the SS-immunopositive neuron profile numbers in the AI and DI+GI of 7 d and 2 m post-SE rats. The regions with decreased numbers of PV-immunopositive neuron profiles overlapped with those where many degenerating cells were detected by Fluoro-Jade B staining. The area of excitatory propagation responding to electrical stimulation of the caudal AI was expanded in 4-7 d post-SE rats, and excitation frequently propagated to the frontal cortex including the motor cortex. Optical signals in the AI of 4-7 d post-SE rats were larger in amplitude than those of controls. In contrast to the AI, the DI of 4-7 d post-SE rats showed similar excitatory propagation pattern and amplitude to that of controls. These results suggest that the region-specific loss of PV-immunopositive neurons occurred in the AI 4-7 d after pilocarpine-induced status epilepticus, which may play an important role in facilitating excitatory propagation in the IC. PMID- 20018233 TI - Potential mechanisms involved in the absorptive transport of cadmium in isolated perfused rabbit renal proximal tubules. AB - Lumen-to-cell transport, cellular accumulation, and toxicity of cadmium as ionic cadmium (Cd(2+)) or as the L-cysteine (Cys) or D,L-homocysteine (Hcy) S-conjugate of cadmium (Cys-S-Cd-S-Cys, Hcy-S-Cd-S-Hcy) were studied in isolated, perfused rabbit proximal tubular segments. When Cd(2+) (0.73 microM) or Cys-S-Cd-S-Cys (0.73 microM) was perfused through the lumen of S(2) segments of the proximal tubule, no visual evidence of cellular pathological changes was detected during 30 min of study. Cd(2+)-transport was temperature-dependent and was inhibited by Fe(2+), Zn(2+), and elevated concentrations of Ca(2+). Luminal uptake of Cys-S-Cd S-Cys was also temperature-dependent and was inhibited by the amino acids L cystine and L-arginine, while stimulated by L-methionine. Neither L-aspartate, L glutamate, the synthetic dipeptide, Gly-Sar nor Zn(2+) had any effect on the rate of Cys-S-Cd-S-Cys transport. CONCLUSIONS: When delivered to the luminal compartment, Cd(2+) appears to be capable of utilizing certain transporter(s) of Zn(2+) and some transport systems sensitive to Ca(2+) and Fe(2+). In addition, Cys-S-Cd-S-Cys and Hcy-S-Cd-S-Hcy appear to be transportable substrates of one or more amino acid transporters participating in luminal absorption of the amino acid L-cystine (such as system b(0,+)). These findings indicate that multiple mechanisms could be involved in the luminal absorption of cadmium (Cd) in proximal tubular segments depending on its form. These findings provide a focus for future studies of Cd absorption in the proximal tubule. PMID- 20018234 TI - Musical experience shapes top-down auditory mechanisms: evidence from masking and auditory attention performance. AB - A growing body of research suggests that cognitive functions, such as attention and memory, drive perception by tuning sensory mechanisms to relevant acoustic features. Long-term musical experience also modulates lower-level auditory function, although the mechanisms by which this occurs remain uncertain. In order to tease apart the mechanisms that drive perceptual enhancements in musicians, we posed the question: do well-developed cognitive abilities fine-tune auditory perception in a top-down fashion? We administered a standardized battery of perceptual and cognitive tests to adult musicians and non-musicians, including tasks either more or less susceptible to cognitive control (e.g., backward versus simultaneous masking) and more or less dependent on auditory or visual processing (e.g., auditory versus visual attention). Outcomes indicate lower perceptual thresholds in musicians specifically for auditory tasks that relate with cognitive abilities, such as backward masking and auditory attention. These enhancements were observed in the absence of group differences for the simultaneous masking and visual attention tasks. Our results suggest that long term musical practice strengthens cognitive functions and that these functions benefit auditory skills. Musical training bolsters higher-level mechanisms that, when impaired, relate to language and literacy deficits. Thus, musical training may serve to lessen the impact of these deficits by strengthening the corticofugal system for hearing. PMID- 20018235 TI - Evaluation of guggulipid and nimesulide on production of inflammatory mediators and GFAP expression in LPS stimulated rat astrocytoma, cell line (C6). AB - AIM OF THE STUDY: The present study was designed to investigate effect of guggulipid, a drug developed by CDRI and nimesulide on LPS stimulated neuroinflammatory changes in rat astrocytoma cell line (C6). MATERIALS AND METHODS: Rat astrocytoma cells (C6) were stimulated with LPS (10 microg/ml) alone and in combinations with different concentrations of guggulipid or nimesulide for 24h of incubation. Nitrite release in culture supernatant, ROS in cells, expressions of COX-2, GFAP and TNF-alpha in cell lysate were estimated. RESULTS: LPS (10 microg/ml) stimulated C6 cells to release nitrite, ROS generation, up regulated COX-2 and GFAP expressions at protein level and TNF-alpha at mRNA level. Both guggulipid and nimesulide significantly attenuated nitrite release, ROS generation and also down regulated expressions of COX-2, GFAP and TNF-alpha. Guggulipid and nimesulide per se did not have any significant effect on C6 cells. CONCLUSION: Results demonstrate the anti-inflammatory effect of guggulipid comparable to nimesulide which suggest potential use of guggulipid in neuroinflammation associated conditions in CNS disorders. PMID- 20018236 TI - Dynamics of telomerase activity in response to acute psychological stress. AB - Telomerase activity plays an essential role in cell survival, by lengthening telomeres and promoting cell growth and longevity. It is now possible to quantify the low levels of telomerase activity in human leukocytes. Low basal telomerase activity has been related to chronic stress in people and to chronic glucocorticoid exposure in vitro. Here we test whether leukocyte telomerase activity changes under acute psychological stress. We exposed 44 elderly women, including 22 high stress dementia caregivers and 22 matched low stress controls, to a brief laboratory psychological stressor, while examining changes in telomerase activity of peripheral blood mononuclear cells (PBMCs). At baseline, caregivers had lower telomerase activity levels than controls, but during stress telomerase activity increased similarly in both groups. Across the entire sample, subsequent telomerase activity increased by 18% one hour after the end of the stressor (p<0.01). The increase in telomerase activity was independent of changes in numbers or percentages of monocytes, lymphocytes, and specific T cell types, although we cannot fully rule out some potential contribution from immune cell redistribution in the change in telomerase activity. Telomerase activity increases were associated with greater cortisol increases in response to the stressor. Lastly, psychological response to the tasks (greater threat perception) was also related to greater telomerase activity increases in controls. These findings uncover novel relationships of dynamic telomerase activity with exposure to an acute stressor, and with two classic aspects of the stress response - perceived psychological stress and neuroendocrine (cortisol) responses to the stressor. PMID- 20018237 TI - ReProGlo: a new stem cell-based reporter assay aimed to predict embryotoxic potential of drugs and chemicals. AB - A stem cell-based reporter assay was developed to detect drug-induced alterations in the canonical Wnt/beta-catenin signaling pathway, which is involved in the regulation of early embryonic development. The so-called ReProGlo assay allows simultaneous determination of cell viability and luciferase reporter activity in a high throughput 96-well microtiter format. A clone of mouse embryonic stem (mES) cells stably expressing the SuperTopFlash reporter was established. This allows Wnt pathway activity determinations in undifferentiated mES cells and their differentiated descendants. Several test chemicals were analyzed in the new assay system. Known embryotoxicants like retinoic acid or lithium chloride induced concentration-dependent increases in reporter activity. The potency of valproic acid and a series of structural analogs to activate the Wnt pathway correlated well with their reported teratogenic activity in the mouse. Cyclophosphamide was also active but only after metabolic activation by hepatocytes. The new test may help to predict embryotoxic potential of chemicals. PMID- 20018238 TI - Host proteins involved in HIV infection: new therapeutic targets. AB - Current treatment of HIV/AIDS consists of a combination of three to five agents targeting different viral proteins, i.e. the reverse transcriptase, protease, integrase and envelope, and aims to suppress viral replication below detectable levels. This "highly active antiretroviral therapy" (HAART) has brought an enormous benefit for life expectancy and quality in HIV-1-infected individuals, at least in industrialized countries. However, significant limitations with regard to efficiency, drug resistance, side effect and costs still exist. Recent data suggest that cellular factors also represent useful targets for therapy. Here, we summarize findings from several genome-wide screens that identified a large number of cellular factors exploited by HIV-1 at each step of its life cycle. Furthermore, we discuss the evidence that humans are equipped with powerful intrinsic defense mechanisms against retroviruses but that HIV-1 has evolved elaborate ways to counteract or evade them. Preventing the use of host cell proteins obligatory for viral replication or strengthening the cellular defense mechanisms may help to reduce viral replication to harmless levels. A better understanding of the host factors that promote or restrict HIV-1 replication may thus lead to the development of novel therapeutics against HIV/AIDS. PMID- 20018239 TI - Physical properties of griseofulvin-lipid nanoparticles in suspension and their novel interaction mechanism with saccharide during freeze-drying. AB - Size reduction of drug particles to the nanoscale is important in improving the dissolution rate of poorly water-soluble drugs. The aim of this study was to investigate the physicochemical properties of griseofulvin (GF)-lipid nanoparticles and the interactions between GF-lipid nanoparticles and various saccharides during freeze-drying. The phase transition temperature of the GF lipid nanoparticle suspension was 56.8 degrees C, whereas that of the lipid nanoparticle suspension alone was 57.9 degrees C, indicating that the GF crystals were incorporated into the lipid phase. The mean particle size of a rehydrated suspension of xylose-containing freeze-dried GF-lipid nanoparticles was about 220 nm. However, the mean particle size on the rehydration of nanoparticles containing mannose (monosaccharide), fructose (disaccharide), lactose (disaccharide), or raffinose (trisaccharide) was about 60 nm, suggesting that these saccharides prevented aggregation during the freeze-drying process. Powder X-ray diffraction revealed that xylose existed in the crystalline state in the freeze-dried nanoparticles, whereas the other saccharides existed in amorphous states. Thus, the crystallization of the saccharide was found to be strongly correlated with the aggregation property of the nanoparticles. In the case of freeze-dried xylose, the nanoparticles were squeezed out as the saccharine crystal lattice arranged itself regularly. Then, the ejected nanoparticles were aggregated. In contrast, in the case of the other freeze-dried saccharide, the saccharide remained incorporated with the GF-lipid nanoparticles because its crystal lattice was arranged irregularly. Thus, the particle size was maintained. PMID- 20018240 TI - Type I collagen promotes epithelial-mesenchymal transition through ILK-dependent activation of NF-kappaB and LEF-1. AB - Collagen I has been shown to promote epithelial-mesenchymal transition (EMT), a critical process of embryonic development and disease progression. However, little is known about the signaling mechanisms by which collagen I induces this cellular transformation. Here we show that collagen I causes ILK-dependent phosphorylation of IkappaB and subsequent nuclear translocation of active NF kappaB, which in turn promotes increased expression of the Snail and LEF-1 transcription factors. ILK also causes inhibitory phosphorylation of GSK-3beta, a kinase that prevents functional activation of both Snail and LEF-1. These transcription factors alter expression of epithelial and mesenchymal markers to initiate EMT and stimulate cell migration. These data provide a foundation for understanding the mechanisms by which collagen I stimulates EMT and identify potential therapeutic targets for suppressing this transition in pathological conditions. PMID- 20018241 TI - Cuticular proteins and seasonal photoperiodism in aphids. AB - For poikilotherm animals such as insects, extreme temperatures can be a severe issue in continental regions. Aphids, which reproduce in spring and summer by viviparity, are prone to death in hard winter conditions. These species exhibit reproductive plasticity adapted to winter by producing oviparous females in autumn, which lay overwintering eggs. This switch is driven by photoperiodism, and long nights are sufficient to trigger the change in reproductive mode. Global transcriptomic analyses applied to the pea aphid Acyrthosiphon pisum for which genomic resources are now available have allowed the identification of several genetic programs regulated by photoperiod shortening. Unexpectedly, one of these genetic programs concerns cuticle proteins and cuticle structure. This opens new tracks for investigations and poses new hypotheses on the link between cuticle modification and neuronal signalisation of photoperiod in aphids in response to seasonal photoperiodism. This review focuses on the description of cuticular protein genes in the pea aphid and their regulation during the change of reproductive mode. PMID- 20018242 TI - Identification of juvenile hormone target genes in the adult female mosquitoes. AB - Many aspects of reproductive maturation in newly emerged adult female mosquitoes are controlled by juvenile hormone (JH). However, the molecular basis of this hormonal regulation remains poorly understood. In this study, we analyzed transcriptome changes in young female adults after topical application of exogenous JH. mRNA levels of 16 and 72 genes were enhanced at 3 h and 12 h after hormone administration, respectively. The observed upregulations can be also induced by the JH analogues, methoprene and pyriproxyfen, but not by farnesol, indicating that these responses are JH-specific. Among the genes activated by JH is the mosquito Kruppel homolog 1 (Kr-h1) gene. Studies in the beetle Tribolium castaneum have indicated that Kr-h1 is transcriptionally activated by JH during metamorphosis and encodes a gene expression regulator acting downstream of the methoprene-tolerant (Met) gene in the JH signaling pathway. We found that the upregulation of AaKr-h1 after eclosion relied on the function of the mosquito Met protein, suggesting that a conserved JH signaling pathway is utilized both in the metamorphosis of T. castaneum and in the adult reproductive maturation of the Aedes aegypti mosquito. This study lays a foundation for a better understanding of the mechanisms of juvenile hormone action. PMID- 20018243 TI - Electron microscopy, immunostaining, cytoskeleton visualization, in situ hybridization, and three-dimensional reconstruction of Xenopus oocytes. AB - Although the overwhelming development of molecular techniques in recent decades has made ultrastructural studies less popular, to the point that ultrastructural interpretation is becoming a dying art, it still remains an indispensable tool for cell and developmental biologists. The introduction of EM-immunocytochemistry and three-dimensional visualization methods allows us to complement the knowledge gained from ultrastructural and molecular approaches. Because the first clues about the functions of newly discovered genes often come from the subcellular localization patterns of their proteins or RNAs, in this chapter we describe the methods that allow for precise ultrastructural localization and visualization of protein and RNA molecules within the compartments, organelles, and cytoskeleton of Xenopus oocytes. PMID- 20018245 TI - The production of soluble and correctly folded recombinant bovine beta lactoglobulin variants A and B in Escherichia coli for NMR studies. AB - The production of soluble and correctly folded eukaryotic proteins in prokaryotic systems has always been hampered by the difference in or lack of cell machinery responsible for folding, post-translation modification and secretion of the proteins involved. In the case of bovine beta-lactoglobulin (BLG), a major cow's milk allergen and a protein widely used for protein folding studies, a eukaryotic yeast expression system has been the preferred choice of many researchers, particularly for the production of isotopically labeled protein required for NMR studies. Although this system yields high amounts of recombinant protein, the BLG produced is usually associated with extracellular polysaccharides, which is problematic for NMR analysis. In our study we show that when co-expressed with the signal-sequence-less disulfide bond isomerase (Delta ssDsbC) in the dual expression vector, pETDUET-1, both BLG A and BLG B can be reproducibly produced in a soluble form. Expression was carried out in Escherichia coli Origami(DE3), a trxB/gor mutant for thioredoxin- and glutathione reductase, which allows for proper formation of disulfide bonds in the cytoplasm. The protein was purified by anion exchange chromatography followed by salting-out at low pH and size exclusion chromatography. Our expression system is able to consistently produce milligram quantities of correctly folded BLG A and B with no additional amino acid residues at the N-terminus, except for a methionine. (15)N-labeled BLG A and B, prepared and purified using this method, produced HSQC spectra typical of native bovine BLG. PMID- 20018244 TI - Quantification of ethanol methyl (1)H magnetic resonance signal intensity following intravenous ethanol administration in primate brain. AB - In vivo(1)H magnetic resonance spectroscopy (MRS) can be used to directly monitor brain ethanol. Previously, studies of human subjects have lead to the suggestion that the ethanol methyl (1)H MRS signal intensity relates to tolerance to ethanol's intoxicating effects. More recently, the ethanol (1)H MRS signal intensity has been recognized to vary between brain gray matter (GM), white matter (WM), and cerebrospinal fluid (CSF) due to differences in T(2) within these environments. The methods presented here extend ethanol MRS techniques to non-human primate subjects. Twelve monkeys were administered ethanol while sedated and positioned within a 3T MRI system. Chemical shift imaging (CSI) measurements were performed following intravenous infusion of 1g/kg ethanol. Magnetic resonance imaging (MRI) data were also recorded for each monkey to provide volume fractions of GM, WM, and CSF for each CSI spectrum. To estimate co variance of ethanol MRS intensity with GM, WM, and CSF volume fractions, the relative contribution of each tissue subtype was determined following corrections for radiofrequency pulse profile non-uniformity, chemical shift artifacts, and differences between the point spread function in the CSI data and the imaging data. The ethanol MRS intensity per unit blood ethanol concentration was found to differ between GM, WM, and CSF. Individual differences in MRS intensity were larger in GM than WM. This methodology demonstrates the feasibility of ethanol MRS experiments and analysis in non-human primate subjects, and suggests GM may be a site of significant variation in ethanol MRS intensity between individuals. PMID- 20018246 TI - A toolbox for ab initio 3-D reconstructions in single-particle electron microscopy. AB - Structure determination of a novel macromolecular complex via single-particle electron microscopy depends upon overcoming the challenge of establishing a reliable 3-D reconstruction using only 2-D images. There are a variety of strategies that deal with this issue, but not all of them are readily accessible and straightforward to use. We have developed a "toolbox" of ab initio reconstruction techniques that provide several options for calculating 3-D volumes in an easily managed and tightly controlled work-flow that adheres to standard conventions and formats. This toolbox is designed to streamline the reconstruction process by removing the necessity for bookkeeping, while facilitating transparent data transfer between different software packages. It currently includes procedures for calculating ab initio reconstructions via random or orthogonal tilt geometry, tomograms, and common lines, all of which have been tested using the 50S ribosomal subunit. Our goal is that the accessibility of multiple independent reconstruction algorithms via this toolbox will improve the ease with which models can be generated, and provide a means of evaluating the confidence and reliability of the final reconstructed map. PMID- 20018247 TI - Protective immunity against viral nervous necrosis (VNN) in brown-marbled grouper (Epinephelus fuscogutattus) following vaccination with inactivated betanodavirus. AB - Viral nervous necrosis (VNN) caused by betanodaviruses has been recently implicated in serious mortalities of groupers in the grow-out culture system. A safe and effective vaccine against this disease is urgently needed. This study demonstrates that a single intramuscular vaccination with formalin-inactivated Philippine strain of piscine betanodavirus (genotype: redspotted grouper nervous necrosis virus; RGNNV) induces potent immune responses and substantial protective immunity against an intramuscular challenge with the homologous virus in brown marbled grouper, Epinephelus fuscogutattus, a highly susceptible marine fish species to VNN. Seroneutralization assay conducted on sera of vaccinated fish revealed the occurrence of substantial neutralizing-antibody titers from Days 15 (mean titer 1:800) to 190 (1:400) with the highest titer observed at Day 60 post vaccination (1:5120). When vaccinated fish were challenged with the homologous virus at Days 15, 30 and 75 post-vaccination, significantly higher survival rates were obtained in these fish compared with their corresponding controls (L-15 injected fish). Abrogation of virus multiplication in all vaccinated survivors was indicated by undetectable virus titers in the brains and kidneys paralleled by significantly high levels of neutralizing antibodies in the sera of these fish. Consecutively, replicates of vaccinated fish that survived betanodavirus challenge at Days 15 and 75 post-vaccination were maintained in flow-through aquaria and rechallenged with the homologous virus 3 and 5 months later, respectively. A significant drop in neutralizing-antibody titers of 3 and 8 folds, respectively, were observed in the sera of Days 15 and 75 post-vaccinated fish assayed before the virus rechallenge. Interestingly, reversion in the levels of neutralizing antibodies to significantly high levels (8-15 folds) were noted in these fish after the virus rechallenge. Taken together, our current data clearly demonstrate that a single administration of the inactivated Philippine strain of betanodavirus vaccine can effectively mount a specific anamnestic response and concomitant long-term protection against VNN in grouper at the grow out culture system. PMID- 20018249 TI - Propranolol reduces emotional distraction in working memory: a partial mediating role of propranolol-induced cortisol increases? AB - Noradrenalin modulates prefrontal function, such as working memory (WM), and is associated with enhanced distractibility, and enhanced memory for emotional events and stimuli. The beta-blocker propranolol has been shown to reduce memory for emotional stimuli. Herein we describe investigations aimed at assessing whether the administration of propranolol would reduce the interference by emotional distractions during WM performance. In a between-subjects design, 48 young, healthy men received 80 mg propranolol (n=25) or placebo (n=23), before performing an "emotional Sternberg task" with neutral and negatively arousing distracters. Compared to placebo, propranolol impaired WM at low load, however, it also reduced the interference by emotional distracters at high load. Furthermore, an explorative moderated-mediation analysis indicated that the observed propranolol effects on emotional distraction were partially mediated by cortisol. In future non-clinical and clinical memory studies using propranolol administration, cortisol elevations should be monitored to further investigate the potential mediating role of cortisol. PMID- 20018248 TI - Trace and contextual fear conditioning is enhanced in mice lacking the alpha4 subunit of the GABA(A) receptor. AB - The GABA(A)R alpha4 subunit is highly expressed in the dentate gyrus region of the hippocampus at predominantly extra synaptic locations where, along with the GABA(A)R delta subunit, it forms GABA(A) receptors that mediate a tonic inhibitory current. The present study was designed to test hippocampus-dependent and hippocampus-independent learning and memory in GABA(A)R alpha4 subunit deficient mice using trace and delay fear conditioning, respectively. Mice were of a mixed C57Bl/6J X 129S1/X1 genetic background from alpha4 heterozygous breeding pairs. The alpha4-knockout mice showed enhanced trace and contextual fear conditioning consistent with an enhancement of hippocampus-dependent learning and memory. These enhancements were sex-dependent, similar to previous studies in GABA(A)R delta knockout mice, but differences were present in both males and females. The convergent findings between alpha4 and delta knockout mice suggests that tonic inhibition mediated by alpha4betadelta GABA(A) receptors negatively modulates learning and memory processes and provides further evidence that tonic inhibition makes important functional contributions to learning and behavior. PMID- 20018252 TI - Inhalation of formaldehyde does not induce genotoxic effects in broncho-alveolar lavage (BAL) cells of rats. AB - Male Fischer-344 rats were exposed to formaldehyde (FA) by inhalation for 4 weeks (6h/day, 5 days/week). Groups of six rats each were exposed to the target concentrations of 0, 0.5, 1, 2, 6, 10 and 15ppm. Potential genotoxic effects in the lung were investigated as part of a comprehensive study on local and systemic toxic and genotoxic effects. Broncho-alveolar lavage (BAL) cells were obtained by lung lavage with physiological saline and counted. From one half of the cells, slides for the micronucleus test (MNT) were prepared by cytocentrifugation; with the other half, the comet assay was performed. DNA migration in the comet assay was measured both directly and after irradiation of the cells with 2Gy gamma radiation. The latter modification of the comet assay was included to increase its sensitivity for the detection of DNA-protein cross-links (DPX). For the comet assay, four slides were analysed from each cell sample, two without and two with irradiation. From each slide, 50 randomly selected cells were measured by image analysis and tail intensity (% tail DNA) and tail moment were evaluated. The frequency of micronucleated BAL cells was determined in acridine orange-stained slides by analysing 2000 cells per animal. FA did not induce any significant effect in any of the genotoxicity tests performed. It can be concluded that inhalation of FA in a 28 days study with FA concentrations up to 15ppm does not lead to genotoxic effects in BAL cells of rats. Because detection of DPX by the comet assay is a very sensitive biomarker of FA exposure of cells, our results suggest that there is no genetically relevant exposure of the lung after FA inhalation. The results of our inhalation study, which was performed under GLP conditions, call into question the biological significance of previously reported genotoxic effects in the lung of rats after FA inhalation. PMID- 20018250 TI - Regenerative stromal cell therapy in allogeneic hematopoietic stem cell transplantation: current impact and future directions. AB - Regenerative stromal cell therapy (RSCT) has the potential to become a novel therapy for preventing and treating acute graft-versus-host disease (GVHD) in the allogeneic hematopoietic stem cell transplant (HSCT) recipient. However, enthusiasm for using RSCT in allogeneic HSCT has been tempered by limited clinical data and poorly defined in vivo mechanisms of action. As a result, the full clinical potential of RSCT in supporting hematopoietic reconstitution and as treatment for GVHD remains to be determined. This manuscript reviews the immunomodulatory activity of regenerative stromal cells in preclinical models of allogeneic HSCT, and emphasizes an emerging literature suggesting that microenvironment influences RSC activation and function. Understanding this key finding may ultimately define the proper niche for RSCT in allogeneic HSCT. In particular, mechanistic studies are needed to delineate the in vivo effects of RSCT in response to inflammation and injury associated with allogeneic HSCT, and to define the relevant sites of RSC interaction with immune cells in the transplant recipient. Furthermore, development of in vivo imaging technology to correlate biodistribution patterns, desired RSC effect, and clinical outcome will be crucial to establishing dose-response effects and minimal biologic dose thresholds needed to advance translational treatment strategies for complications like GVHD. PMID- 20018251 TI - The co-chaperone BAG3 interacts with the cytosolic chaperonin CCT: new hints for actin folding. AB - It has been recently hypothesized that BAG3 protein, a co-chaperone of Hsp70/Hsc70, is involved in the regulation of several cell processes, such as apoptosis, autophagy and cell motility. Following the identification of Hsc70/Hsp70, further BAG3 molecular partners such as PLC-gamma and HspB8 were likewise identified, thus contributing to the characterization of the mechanisms and the biological roles carried out by this versatile protein. By using a His tagged BAG3 protein as bait, we fished out and identified the cytosolic chaperonin CCT, a new unreported BAG3 partner. The interaction between BAG3 and CCT was confirmed and characterized by co-immunoprecipitation experiments and surface plasmon resonance techniques. Furthermore, our analyses showed a slower CCT association and a faster dissociation with a truncated form of BAG3 containing the BAG domain, thus indicating that other protein regions are essential for a high-affinity interaction. ATP or ADP does not seem to significantly influence the chaperonin binding to BAG3 protein. On the other hand, our experiments showed that BAG3 silencing by small interfering RNA slowed down cell migration and influence the availability of correctly folded monomeric actin, analyzed by DNAse I binding assays and latrunculin A depolymerization studies. To our knowledge, this is the first report showing a biologically relevant interaction between the chaperonin CCT and BAG3 protein, thus suggesting interesting involvement in the folding processes regulated by CCT. PMID- 20018253 TI - FociCounter: a freely available PC programme for quantitative and qualitative analysis of gamma-H2AX foci. AB - Gamma-H2AX foci are sensitive and specific indicator for the induction of DNA double-strand breaks (DSBs) and an immunocytochemical assay with antibodies recognizing gamma-H2AX has become the gold standard for the detection of this type of DNA lesion. Quantification of gamma-H2AX foci can be achieved by various methods such as Western blotting, flow cytometry, visual analysis and computational analysis with a fluorescence microscope. The best sensitivity is achieved by computer analysis. Since no freeware programme for the analysis of gamma-H2AX foci exists for a PC platform, the aim of our study was to develop a simple and user-friendly public-domain software. The algorithm applied in our programme allows determination of the number of foci in a single cell, a focus intensity per cell, as well as a cell intensity. Its graphical user interface is based on a GTK+ library and the whole application can be run under a variety of operating systems, including MS Windows and Linux. The programme called FociCounter is publicly available at http://focicounter.sourceforge.net. Application of the programme was tested by analysing gamma-H2AX foci in CHO and MO59K cells irradiated in vitro with X-rays and validated by comparing the results obtained with the outcome of automated image analysis and flow cytometry. PMID- 20018255 TI - Systematics of Mepraia (Hemiptera-Reduviidae): cytogenetic and molecular variation. AB - The haematophagous insects of the subfamily Triatominae (Hemiptera-Reduviidae) have great epidemiological importance as vectors of Trypanosoma cruzi, the causative agent of Chagas disease. Mepraia was originally described as a monotypic genus comprised of Mepraia spinolai, distributed along coastal areas of northern Chile (from Region I to the Metropolitan Region). Recently, some M. spinolai populations have been ranked as a new species named Mepraia gajardoi. Several populations along the distribution range of the genus were sampled, and genetic differentiation was studied based upon the analysis of three molecular markers: cytogenetics (karyotype and chromosome behaviour during meiosis using the C-banding technique), mitochondrial DNA (a cytochrome oxidase I gene fragment), and nuclear ribosomal DNA (intergenic region including the two internal transcribed spacers ITS-1 and ITS-2 and the 5.8S rRNA gene). The data here presented indicate that populations within the Mepraia genus (excluding Region II specimens) can be divided into two separate lineages. One lineage is comprised of specimens from the northernmost Region I and represents M. gajardoi. The other includes samples from the southern III, IV and the Metropolitan Regions, and represents M. spinolai. Region II individuals deserve particular attention as their relationship to the two identified lineages is not clear-cut. While they appear to belong to M. spinolai based on cytogenetics and rDNA markers, COI results indicate a closer relationship to M. gajardoi. This disagreement can be due to mitochondrial DNA introgression or the retention of ancestral polymorphisms. PMID- 20018256 TI - Low genetic diversities of rabies virus populations within different hosts in Brazil. AB - The low rates of nonsynonymous evolution observed in natural rabies virus (RABV) isolates are suggested to have arisen in association with the structural and functional constraints operating on the virus protein and the infection strategies employed by RABV within infected hosts to avoid strong selection by the immune response. In order to investigate the relationship between the genetic characteristics of RABV populations within hosts and the virus evolution, the present study examined the genetic heterogeneities of RABV populations within naturally infected dogs and foxes in Brazil, as well as those of bat RABV populations that were passaged once in suckling mice. Sequence analyses of complete RABV glycoprotein (G) genes showed that RABV populations within infected hosts were genetically highly homogeneous whether they were infected naturally or experimentally (nucleotide diversities of 0-0.95x10(-3)). In addition, amino acid mutations were randomly distributed over the entire region of the G protein, and the nonsynonymous/synonymous rate ratios (d(N)/d(S)) for the G protein gene were less than 1. These findings suggest that the low genetic diversities of RABV populations within hosts reflect the stabilizing selection operating on the virus, the infection strategies of the virus, and eventually, the evolutionary patterns of the virus. PMID- 20018257 TI - Demethoxycurcumin, a natural derivative of curcumin attenuates LPS-induced pro inflammatory responses through down-regulation of intracellular ROS-related MAPK/NF-kappaB signaling pathways in N9 microglia induced by lipopolysaccharide. AB - Our previous report has showed that demethoxycurcumin (DMC), a natural derivative of curcumin (Cur), exhibited stronger inhibitory activity on nitric oxide (NO) and tumor necrosis factor-alpha (TNF-alpha) production compared with Cur in lipopolysaccharide (LPS) activated rat primary microglia. In the present study, the effect and possible mechanism of DMC on the production of pro-inflammatory mediators in LPS-activated N9 microglial cells were further investigated. The results showed that DMC significantly suppressed the NO production induced by LPS in N9 microglial cells through inhibiting the protein and mRNA expression of inducible NO synthase (iNOS). DMC also decreased LPS-induced TNF-alpha and IL 1beta expression at both transcriptional and protein level in a concentration dependent manner. Further studies revealed that DMC blocked IkappaBalpha phosphorylation and degradation, inhibited the phosphorylation of mitogen activated protein kinases (MAPKs). Moreover, the level of intracellular reactive oxygen species (iROS) was significantly increased by LPS, which is mainly mediated by the up-regulated expression of gp91phox, the catalytic subunit of nicotinamide adenine dinucleotide phosphate reduced (NADPH) oxidase. Both DMC and Cur could markedly decrease iROS production and the expression of NADPH oxidase induced by LPS, with more potent inhibitory activity of DMC. In summary, these data suggest that DMC exerts its in vitro anti-inflammatory effect in LPS activated N9 microglial cells by blocking nuclear factor-kappaB (NF-kappaB) and MAPKs activation, which may be partly due to its potent down-regulation of the NADPH-derived iROS production. PMID- 20018258 TI - Relevance of quantitative measurements of anti-Abeta antibodies in therapeutic intravenous immunoglobulin using synthetic peptides. PMID- 20018260 TI - Porous diopside (CaMgSi(2)O(6)) scaffold: A promising bioactive material for bone tissue engineering. AB - Diopside (CaMgSi(2)O(6)) powders and dense ceramics have been shown to be bioactive biomaterials for bone repair. The aim of this study is to prepare bioactive diopside scaffolds and examine their physicochemical and biological properties. X-ray diffraction, scanning electron microscopy (SEM), micro computerized tomography and energy-dispersive spectrometry were used to analyse the composition, microstructure, pore size and interconnectivity of the diopside scaffolds. The mechanical strength and stability as well as the degradation of the scaffolds were investigated by testing the compressive strength, modulus and silicon ions released, respectively. Results showed that highly porous diopside scaffolds with varying porosity and high interconnectivity of 97% were successfully prepared with improved compressive strength and mechanical stability, compared to the bioglass and CaSiO(3) scaffolds. The bioactivity of the diopside scaffolds was assessed using apatite-forming ability in simulated body fluids (SBF) and by their support for human osteoblastic-like cell (HOB) attachment, proliferation and differentiation using SEM, and MTS and alkaline phosphatase activity assays, respectively. Results showed that diopside scaffolds possessed apatite-forming ability in SBF and supported HOB attachment proliferation and differentiation. Bioactive diopside scaffolds were prepared with excellent pore/structure art, and improved mechanical strength and mechanical stability, suggesting their possible applications for bone tissue engineering regeneration. PMID- 20018259 TI - (D)-Amino acid analogues of DT-2 as highly selective and superior inhibitors of cGMP-dependent protein kinase Ialpha. AB - The cGMP-dependent protein kinase type I (PKG I) is an essential regulator of cellular function in blood vessels throughout the body. DT-2, a peptidic inhibitor of PKG, has played a central role in determining the molecular mechanisms of vascular control involving PKG and its signaling partners. Here, we report the development of (d)-amino acid DT-2 derivatives, namely the retro inverso ri-(d)-DT-2 and the all (d)-amino acid analog, (d)-DT-2. Both peptide analogs were potent PKG Ialpha inhibitors with K(i) values of 5.5 nM (ri-(d)-DT 2) and 0.8 nM ((d)-DT-2) as determined using a hyperbolic mixed-type inhibition model. Also, both analogs were proteolytically stable in vivo, showed elevated selectivity, and displayed enhanced membrane translocation properties. Studies on isolated arteries from the resistance vasculature demonstrated that intraluminally perfused (d)-DT-2 significantly inhibited vasodilation induced by 8-Br-cGMP. Furthermore, in vivo application of (d)-DT-2 established a uniform translocation pattern in the resistance vasculature, with exception of the brain. Thus, (d)-DT-2 caused significant increases in mean arterial blood pressure in unrestrained, awake mice. Further, mesenteric arteries isolated from (d)-DT-2 treated animals showed a markedly reduced dilator response to 8-Br-cGMP in vitro. Our results clearly demonstrate that (d)-DT-2 is a superior inhibitor of PKG Ialpha and its application in vivo leads to sustained inhibition of PKG in vascular smooth muscle cells. The discovery of (d)-DT-2 may help our understanding of how blood vessels constrict and dilate and may also aid the development of new strategies and therapeutic agents targeted to the prevention and treatment of vascular disorders such as hypertension, stroke and coronary artery disease. PMID- 20018261 TI - Tribological behavior of DLC-coated articulating joint implants. AB - Coatings from diamond-like carbon (DLC) have been proven to be an excellent choice for wear reduction in many technical applications. However, for successful adaption to the total joint replacement field, layer performance, stability and adhesion in realistic physiological setups are quintessential and these aspects have not been consistently researched. In our team's efforts to develop long-term stable DLC implant coatings, test results gained from a simplified linear spinal simulator setup are presented. It is shown that metal-on-metal (MoM) pairs perform well up to 7 million loading cycles, after which they start to generate wear volumes in excess of 20 times those of DLC-coated implants. This is attributed to the roughening observed on unprotected metal surfaces. Furthermore, we illustrate that in contrast to DLC-on-DLC, MoM tribopairs require protein containing media to establish low-friction conditions. Finally, results of defect monitoring during testing are presented, showing catastrophic failure of layers whose interfaces are too weak with respect to the stress-corrosion-cracking mechanism encountered in vivo. PMID- 20018262 TI - Electrodeposition of Ca-P coatings on biodegradable Mg alloy: in vitro biomineralization behavior. AB - Preparing stabilized apatite on biodegradable Mg alloy may improve biocompatibility and promote osteointegration. In the present work, three kinds of Ca-P coatings, brushite (DCPD, CaHPO(4).2H(2)O), hydroxyapatite (HA, Ca(10)(PO(4))(6)(OH)(2)) and fluoridated hydroxyapatite (FHA, Ca(5)(PO(4))(3)(OH)(1-)(x)F(x)) are fabricated by electrodeposition on a biodegradable Mg-Zn alloy. The crystalline structures, morphologies and compositions of these Ca-P coatings have been characterized by X-ray diffrection, scanning electron microscopy and energy-dispersive spectoscopy. The effects of these coatings on the degradation behavior and mineralization activity of the Mg Zn alloy have also been investigated. The experimental results showed that these coatings decreased the degradation rate of Mg-Zn alloy, while the precipitates on the uncoated and DCPD-coated Mg-Zn alloy in modified simulated biological fluid had low Ca/P molar ratios, which delayed bone-like apatite formation. Both the HA and FHA coating could promote the nucleation of osteoconductive minerals (bone like apatite or beta-TCP) for 1month. However, the HA coating transformed from DCPD through alkali heat treatment was fragile and less stable, and therefore its long-term corrosion resistance was not satisfactory. Instead, the FHA was more stable and had better corrosion resistance, and thus it should be better suited as a coating of Mg implants for orthopedic applications. PMID- 20018263 TI - Optimized enzymatic synthesis of caffeic acid phenethyl ester by RSM. AB - In this study, optimization of enzymatic synthesis of caffeic acid phenethyl ester (CAPE), catalyzed by immobilized lipase (Novozym 435) from Candida antarctica was investigated. Novozym 435 was used to catalyze caffeic acid and 2 phenylethanol in an isooctane system. Response surface methodology (RSM) and 5 level-4-factor central-composite rotatable design (CCRD) were employed to evaluate the effects of synthesis parameters, such as reaction temperature (30-70 degrees C), reaction time (24-72 hours), substrate molar ratio of caffeic acid to 2-phenylethanol (1:10-1:90) and enzyme amounts (100-500 PLU) on percentage conversion of CAPE by direct esterification. Reaction temperature and time had significant effects on percent conversion. On the basis of ridge max analysis, the optimum conditions for synthesis were: reaction time 59 hours, reaction temperature 69 degrees C, substrate molar ratio 1:72 and enzyme amount 351 PLU. The molar conversion of predicted values and actual experimental values were 91.86+/-5.35% and 91.65+/-0.66%, respectively. PMID- 20018264 TI - Measles in Papua New Guinea: an age-specific serological survey. AB - We aimed to determine the proportion of the population in Madang (Papua New Guinea) immune to measles infection by age groups, with respect to immunization status and study location, using dried blood sampling technology. We performed a prospective cross-sectional sero-survey. Population immunity against measles was sub-optimal (77%) and reported measles vaccine coverage in children <10 years of age was low (41%). The urban population was more susceptible to measles infection, compared with the rural population (66% vs 79% immune, aOR=0.6, p=0.05). Sero-conversion and long term protection rates appeared to be higher when at least one dose of vaccine was provided at or after 12 months of age (84% vs 59%, aOR=4.3, p=0.004). Such a dose is, however, not currently prescribed by the national immunization schedule. PMID- 20018265 TI - Seasonal influenza vaccine elicits heterosubtypic immunity against H5N1 that can be further boosted by H5N1 vaccination. AB - Recent findings indicate that seasonal influenza vaccination or infection of healthy humans may contribute to heterosubtypic immunity against new influenza A subtypes, such as H5N1. Here, we investigated whether seasonal influenza vaccination in a mouse model could induce any immunity against the H5N1 subtype. It could be demonstrated that, largely due to the H1N1 component strain A/NewCaledonia/20/99, parenteral immunization of mice with a trivalent seasonal influenza vaccine elicited heterosubtype H5-reactive antibodies able to confer partial protection against H5N1 influenza virus infection. Furthermore, the trivalent seasonal influenza vaccine was found to be compatible with a whole virus H5N1 vaccine in a heterologous prime-boost immunization regimen, achieving superior efficacy compared to a single immunization with an equivalent low-dose of the H5N1 vaccine. PMID- 20018266 TI - Differential ability of novel attenuated targeted deletion mutants of Francisella tularensis subspecies tularensis strain SCHU S4 to protect mice against aerosol challenge with virulent bacteria: effects of host background and route of immunization. AB - Francisella tularensis subspecies tularensis is a highly virulent facultative intracellular pathogen of humans and a potential biological weapon. A live vaccine strain, F. tularensis LVS, was developed more than 50 years ago by pragmatic attenuation of a strain of the less virulent holarctica subspecies. LVS was demonstrated to be highly effective in human volunteers who were exposed to intradermal challenge with fully virulent subsp. tularensis, but was less effective against aerosol exposure. LVS faces regulatory hurdles that to date have prevented its licensure for general use. Therefore, a better defined and more effective vaccine is being sought. To this end we have created gene deletion mutants in the virulent subsp. tularensis strain and tested them for their ability to elicit a protective immune response against systemic or aerosol challenge with the highly virulent wild-type subsp. tularensis strain, SCHU S4. Both oral and intradermal (ID) primary vaccination routes were assessed in BALB/c and C3H/HeN mice as was oral boosting. One SCHU S4 mutant missing the heat shock gene, clpB, was significantly more attenuated than LVS whereas a double deletion mutant missing genes FTT0918 and capB was as attenuated as LVS. In general mice immunized with SCHU S4DeltaclpB were significantly better protected against aerosol challenge than mice immunized with LVS. A single ID immunization of BALB/c mice with SCHU S4DeltaclpB was at least as effective as any other regimen examined. Mice immunized with SCHU S4Delta0918DeltacapB were generally protected to a similar degree as mice immunized with LVS. A preliminary examination of immune responses to vaccination with LVS, SCHU S4DeltaclpB, or SCHU S4Delta0918DeltacapB provided no obvious correlate to their relative efficacies. PMID- 20018267 TI - Identification of protective antigens by RNA interference for control of the lone star tick, Amblyomma americanum. AB - The lone star tick, Amblyomma americanum, vectors pathogens of emerging diseases of humans and animals in the United States. Currently, measures are not available for effective control of A. americanum infestations. Development of vaccines directed against tick proteins may reduce tick infestations and the transmission of tick-borne pathogens. However, the limiting step in tick vaccine development has been the identification of tick protective antigens. Herein, we report the application of RNA interference (RNAi) for screening an A. americanum cDNA library for discovery of tick protective antigens that reduce tick survival and weights after feeding. Four cDNA clones, encoding for putative threonyl-tRNA synthetase (2C9), 60S ribosomal proteins L13a (2D10) and L13e (2B7), and interphase cytoplasm foci protein 45 (2G7), were selected for vaccine studies in cattle, along with subolesin, a tick protective protein identified previously. In vaccinated cattle, an overall efficacy (E)>30% was obtained when considering the vaccine effect on both nymphs and adults, but only 2D10, 2G7 and subolesin affected both tick stages. The highest efficacy of control for adult ticks (E>55%) was obtained in cattle vaccinated with recombinant 2G7 or subolesin. These collective results demonstrated the feasibility of developing vaccines for the control of lone star tick infestations. The use of RNAi for identification of tick protective antigens proved to be a rapid and cost-effective tool for discovery of candidate vaccine antigens, and this approach could likely be applied to other parasites of veterinary and medical importance. PMID- 20018268 TI - Cytokine content in lymphoid and white adipose tissues after repeated CpG oligodeoxynucleotide administration in trained rats. AB - The increased threat of bioterrorism and the emergence of potentially fatal diseases underscores the need to improve treatments for protecting all segments of the human population including military personnel. New methods need to be developed. The ability of oligodeoxynucleotides containing the CpG motif (CpG ODNs) to promote the production of T(H)1-type pro-inflammatory cytokines suggest they might be useful as vaccine adjuvants, but their potential effects during exercise have not been widely studied. Repeated administration of CpG ODN in sedentary rats promoted the production of T(H)1-type pro-inflammatory cytokines in spleen, Peyer's patches and adipose tissues. However, such an increase was not observed in trained rats, suggesting that CpG would not be the best agent for vaccine adjuvants and immunomodulation in intensely trained rats. PMID- 20018269 TI - Construction and immunogenicity of pseudotype baculovirus expressing Toxoplasma gondii SAG1 protein in BALB/c mice model. AB - Toxoplasma gondii is a protozoan parasite causing toxoplasmosis to almost one third of population all over the world. One of the most efficient ways to control this disease is immunization. However, so far, there is no effective vaccine available against this pathogen. Recently, a baculovirus pseudotype with vesicular stomatitis virus G protein (Bac-VSV-G) was found to efficiently transduce and express transgenes on mammalian cells, so it was considered as an excellent expressing vector. In this study, the value of Bac-VSV-G in delivering T. gondii antigen was investigated. T. gondii SAG1 gene was cloned into Bac-VSV G, and recombinant baculovirus BV-G-SAG1 was obtained. Indirect immunofluorescence test showed BV-G-SAG1 was efficiently transduced and expressed in pig kidney cells. Then BALB/c mice were immunized with BV-G-SAG1 at different doses (1 x 10(8), 1 x 10(9), and 1 x 10(10)PFU/mouse) and challenged with T. gondii RH strain tachyzoites after immunization. The levels of specific T. gondii antibody, interferon (IFN)-gamma, IL-4, IL-10 expression and release, and the survival rate of treated mice were evaluated. Compared with the mice immunized with DNA vaccine (pcDNA/SAG1) encoding the same gene, BV-G-SAG1 induced higher levels of specific T. gondii antibody and (IFN)-gamma expression with dose dependent manner and the survival rate of mice with BV-G-SAG1 was significantly improved. These results indicated that pseudotype baculovirus-mediated gene delivery can be utilized as an alternative strategy to develop new generation of vaccines against T. gondii infection. PMID- 20018270 TI - Would an in vitro ELISA test be a suitable alternative potency method to the in vivo immunogenicity assay commonly used in the context of international Hepatitis A vaccines batch release? AB - Since many years Afssaps applies the 3R's strategy (replacement, reduction and refinement) for the use of laboratory animal testing in the framework of vaccine batch release. In this context, for Hepatitis A vaccines, a study was carried out to assess the feasibility of replacing the in vivo "Gold Standard" potency assay by the Afssaps' validated in-house antigen content in vitro assay on routine testing. The use of a panel of potent vaccine batches and reduced-potency samples by heating demonstrated a correlation between the two methods. This encourages Afssaps to progressively switch from in vivo to in vitro assay in the framework of Hepatitis A vaccines batch release. PMID- 20018271 TI - Comparison of different prime-boost regimes with DNA and recombinant Orf virus based vaccines expressing glycoprotein D of pseudorabies virus in pigs. AB - Both DNA and Orf virus (ORFV; Parapox virus) based vaccines have shown promise as alternatives for conventional vaccines in pigs against pseudorabies virus (PRV) infection causing Aujeszky's disease. In the present study we evaluated the efficacy of different prime-boost regimes in pigs in terms of immunogenicity and protection against challenge infection with PRV. The different prime-boost regimes consisted of the homologous prime-boost regimes (DNA followed by DNA or ORFV followed by ORFV) and the heterologous prime-boost regimes (DNA followed by ORFV and ORFV followed by DNA), all based on glycoprotein D (gD) of PRV. Moreover, we compared the efficacy of the different prime-boost regimes with the efficacy of a conventional modified live vaccine (MLV). The different prime-boost regimes resulted in different levels of immunity and protection against challenge infection. Most effective was the regime of priming with DNA vaccine followed by boosting with the ORFV based vaccine. This regime resulted in strong antibody responses, comparable to the antibody responses obtained after prime-boost vaccination with a conventional MLV vaccine. Also with regard to protection, the prime DNA-boost ORFV regime performed better than the other prime-boost regimes. This study demonstrates the potential of a heterologous prime-boost vaccination strategy against PRV based on a single antigen, and that in the natural host, the pig. PMID- 20018272 TI - The effects of sex, age, and interval duration on the perception of time. AB - The present experiment examined the interactive effects of sex, age, and interval duration on individual's time perception accuracy. Participants engaged in the duration production task and subsequently completed questionnaires designed to elicit their temporal attitudes. The overall group of 100 individuals was divided evenly between the sexes. Five groups, each composed of 10 males and 10 females, were divided by decades of age ranging from 20 to 69 years old. The specific time estimation task was an empty interval production procedure composed of 50 trials on each of four different intervals of 1, 3, 7, and 20 s, respectively. The presentation orders of these intervals were randomized across participants but yoked across the sexes within each of the respective age groups. Analysis of the production results indicated significant influences for the sex of the participant while age did not appear to affect estimates of these short durations. Temporal attitudes, as reflected in responses to time questionnaire inquiries, did however exhibit significant differences across age. The contending theoretical accounts of such sex and age differences are considered and explanatory accounts that present a synthesis of endogenous and exogenous causal factors are discussed in light of the present pattern of findings. PMID- 20018273 TI - Presence and quantification of mast cells in the gingiva of cats with tooth resorption, periodontitis and chronic stomatitis. AB - OBJECTIVE: Mast cells are tissue-dwelling granule-containing immune cells that play a pivotal role in the pathogenesis of inflammation and other processes. The three most common orodental disorders in cats are periodontitis, feline resorptive lesions (FRL), and chronic gingivostomatitis (FCGS). The presence and density of mast cells in the gingiva has been established in healthy cats but not in cats affected by FRL, FCGS or periodontitis. The aim of the present study was to document and quantify the presence of mast cells in the gingiva adjacent to teeth affected by FRL, FCGS or chronic periodontitis. METHODS: Samples from the gingiva of 32 cats affected by FRL, FCGS or periodontitis were obtained and compared to samples obtained from 7 specific pathogen-free (SPF) cats. Evaluation of mast cells and the inflammatory infiltrate were performed on hematoxylin and eosin, and toluidine blue stained sections. RESULTS: Mast cells densities were significantly increased in gingival tissues adjacent to teeth affected by FRL, FCGS or periodontitis in comparison to SPF samples. There were no significant differences between gingival tissues of the FRL, FCGS and periodontitis groups. However, the relative inflammatory score in the FRL group was significantly lower as compared to the FCGS or periodontitis groups, yet with similar density of mast cells. CONCLUSION: In the gingiva of cats affected with FRL, FCGS or periodontitis, there is an increase in the number of mast cells. The high number of mast cells in the FRL group and concurrent mild inflammatory reaction suggests the notion that mast cells may potentially play role in the pathogenesis of FRL. PMID- 20018274 TI - Relationships of beta-blockers and anxiolytics intake and salivary secretion, masticatory performance and taste perception. AB - OBJECTIVE: Assess the influence of salivary flow on physiological parameters of the stomatognathic system in patients who take beta-blockers or anxiolytic medications. DESIGN: Sixty patients were divided into three groups based on the following criteria: Group 1, control (n=20; no use of medication); Group 2, use of antihypertensive beta-blockers (n=20); and Group 3, use of benzodiazepine anxiolytics (n=20). Salivary flow was assessed by determining stimulated and non stimulated flow/minute. The quantification of the sense of taste was determined on a visual analogue scale (VAS) using solutions of 0.9% NaCl (salty), 50% sucrose (sweet), 20% unsweetened coffee (bitter) and 4.2% vinegar (sour). The DMFT index (number of decayed/missing/filled teeth) was determined by a calibrated examination, following the criteria of the World Health Organization (WHO). Masticatory performance was assessed with an Optosil comminution test and Rosim-Ramler equation. RESULTS: The results did not reveal a significant correlation between salivary flow and masticatory performance (p>0.05). We observed significant decreased non-stimulated salivary flow for Group 2 (p=0.05) when compared to controls. However, taste perception was not influenced by salivary secretion amongst groups. Furthermore, we observed a significant negative correlation between non-stimulated salivary flow and DMFT in Group 1 (p=0.02; r=-0.52). CONCLUSIONS: Patients under beta-blockers therapy presented reduced non-stimulated salivary flow when compared to controls, without influencing the sense of taste or masticatory performance. The use of anxiolytics did not affect salivary flow and taste perception in the studied sample. PMID- 20018275 TI - Unintentional memory inhibition is weakened in non-clinical dissociators. AB - Although participants with dissociation proneness showed inefficient cognitive inhibition, whether and under what stimulus-task contexts dissociators show inefficient memory inhibition remains inconclusive. This study investigated the relationship between trait dissociation and basic operation of memory control using a non-clinical sample. To reduce the involvement of strategic control and the influence of emotionality, the retrieval-practice paradigm was adopted to examine unintentional memory inhibition of neutral materials. Both the low- and middle-dissociation groups showed the forgetting effect, resulting from suppressing competing memories while retrieving a target. In contrast, the high dissociation group did not show the forgetting effect although their performance in the baseline condition and in recalling practiced items was comparable to the other two groups. High dissociation proneness is linked with weakened memory inhibition that may cause diverse memory problems in clinical patients. PMID- 20018276 TI - Motion through syntactic frames. AB - The introduction of Talmy's (1985, 2000) typology sparked significant interest in linguistic relativity in the arena of motion language. Through careful analysis of the conflation patterns evident in the language of motion events, Talmy noted that one class of languages, V-languages, tends to encode path along with the fact of motion in motion verbs, while a second class, S-languages, tends to encode manner. In the experimental literature, it was reasoned that speakers may be expected to extend novel verbs in accordance with the lexicalization patterns of their native languages. However, the results regarding this prediction are mixed. In this paper, I examine the interplay between the meaning encoded in the motion verb itself and the meaning encoded in the motion description construction, offering a Gricean explanation for co-occurrence patterns and, by extension, for the mixed results. I then explore the implications of this argument for research on possible language effects on thought in this domain. PMID- 20018277 TI - Fast FFT-based bioheat transfer equation computation. AB - This paper describes a modeling method of the tissue temperature evolution over time in hyper or hypothermia. The tissue temperature evolution over time is classically described by Pennes' bioheat transfer equation which is generally solved by a finite difference method. In this paper we will present a method where the bioheat transfer equation can be algebraically solved after a Fourier transformation over the space coordinates. As an example, we implemented this method for the simulation of a percutaneous high intensity ultrasound hepatocellular carcinoma curative treatment and compared it with the finite difference method and experimental data. PMID- 20018278 TI - Correlates of serum dioxin to self-reported exposure factors. AB - The aim of the current analysis was to examine the determinates of lipid-adjusted body levels of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) from occupational histories, age, body mass index, and self-reported information from a questionnaire. We collected serum from 346 workers at a New Zealand chemical plant that manufactured and formulated the herbicide, 2,4,5 trichlorophenoxyacetic acid (2,4,5-T). Age, body mass index, and employment history were significant determinates of TCDD. The self-reported data on occupation, residence, and general diet were not predictive of serum levels and we observed no evidence of increased TCDD levels from living close to the site. For participants with putative occupational exposure, employment history and personal factors were important to understand the range of TCDD serum levels. For employees without direct occupational exposure, and resulting lower dioxin levels, we recommend further efforts to develop and validate questionnaires to better evaluate environmental sources of dioxins. PMID- 20018279 TI - Endoscopic closure of gastrogastric fistulas by using a tissue apposition system (with videos). AB - BACKGROUND: Gastrogastric fistulas (GGFs) are seen in 1.5% to 12.5% of patients after Roux-en-Y gastric bypass (RYGB) bariatric surgery, often leading to failure to lose adequate weight. OBJECTIVE: The aim of this study was to assess the feasibility, safety, and percentage of successful primary endoluminal closures of GGFs by using a recently developed tissue apposition system in combination with local mucosectomy. DESIGN: A feasibility and outcome study following institutional review board protocol. SETTING: Tertiary referral teaching hospital, Legacy Health System, Portland, Oregon. INTERVENTIONS: A combination of mucosectomy and nonresorbable tissue apposition is used to achieve a permanent closure of the GGF. PATIENTS: Four patients with 5 GGFs after RYGB; the mean fistula diameter of was 18.6 mm (range 10-30 mm). RESULTS: Primary closure rate (1 endoscopic session) of 5 GGFs was 100%. The mean procedure time was 88.5 minutes. One to 4 pairs of tissue anchors were used to close the fistulas. The mean time for performing mucosectomy was 21.6 minutes (range 8-42 minutes) and 39.6 minutes (range 12-58 minutes) for fistula closure. Estimated blood loss was on average 2 mL (range 0-5 mL). No complications were recorded. Early success (3 months), as evidenced by early satiety and weight loss, was noted for 3 of 4 patients. After 3 months, only the smallest fistula (10 mm) was still completely closed, and after 6 months, it also showed a pinhole opening. CONCLUSION: It was feasible to close all fistulas endoscopically without complications. Permanent closure of GGFs could not be achieved. PMID- 20018280 TI - A retrograde-viewing device improves detection of adenomas in the colon: a prospective efficacy evaluation (with videos). AB - BACKGROUND: Colonoscopy may fail to detect neoplasia located on the proximal sides of haustral folds and flexures. The Third Eye Retroscope (TER) provides a simultaneous retrograde view that complements the forward view of a standard colonoscope. OBJECTIVE: To evaluate the added benefit for polyp detection during colonoscopy of a retrograde-viewing device. DESIGN: Open-label, prospective, multicenter study evaluating colonoscopy by using a TER in combination with a standard colonoscope. SETTING: Eight U.S. sites, including university medical centers, ambulatory surgery centers, a community hospital, and a physician's office. PATIENTS: A total of 249 patients (age range 55-80 years) presenting for screening or surveillance colonoscopy. INTERVENTIONS: After cecal intubation, the disposable TER was inserted through the instrument channel of the colonoscope. During withdrawal, the forward and retrograde video images were observed simultaneously on a wide-screen monitor. MAIN OUTCOME MEASUREMENTS: The number and sizes of lesions (adenomas and all polyps) detected with the standard colonoscope and the number and sizes of lesions found only because they were first detected with the TER. RESULTS: In the 249 subjects, 257 polyps (including 136 adenomas) were identified with the colonoscope alone. The TER allowed detection of 34 additional polyps (a 13.2% increase; P < .0001) including 15 additional adenomas (an 11.0% increase; P < .0001). For lesions 6 mm or larger, the additional detection rates with the TER for all polyps and for adenomas were 18.2% and 25.0%, respectively. For lesions 10 mm or larger, the additional detection rates with the TER for all polyps and for adenomas were 30.8% and 33.3%, respectively. In 28 (11.2%) individuals, at least 1 additional polyp was found with the TER. In 8 (3.2%) patients, the polyp detected with the TER was the only one found. Every polyp that was detected with the TER was subsequently located with the colonoscope and removed. For all polyps and for adenomas, the additional detection rates for the TER were 9.7%/4.1% in the left colon (the splenic flexure to the rectum) and 16.5%/14.9% in the right colon (the cecum to the transverse colon), respectively. LIMITATIONS: There was no randomization or comparison with a separate control group. CONCLUSIONS: A retrograde-viewing device revealed areas that were hidden from the forward-viewing colonoscope and allowed detection of 13.2% additional polyps, including 11.0% additional adenomas. Additional detection rates with the TER for adenomas 6 mm or larger and 10 mm or larger were 25.0% and 33.3%, respectively. ( CLINICAL TRIAL REGISTRATION NUMBER: NCT00657371.). PMID- 20018281 TI - Mannitol as salvage treatment for Complex Regional Pain Syndrome Type I. AB - INTRODUCTION: Complex Regional Pain Syndrome Type I (CRPS I) is a continuation of symptoms and signs due to a pathological exaggerated reaction in an extremity of the human body after an injury or operation. Although the clinical picture of CRPS I in the majority of patients is well known, the underlying pathophysiology remains unclear. In The Netherlands, intravenous mannitol administration used as hydroxyl radical scavenger for patients who do not respond to conservative treatment of CRPS I is advocated but little evidence supports this salvage strategy. In this study the effect of mannitol as salvage medication was evaluated in a well-defined multimodal step-up treatment protocol. PATIENTS AND METHODS: A consecutive group of 68 adult patients with persistent CRPS I was analysed, who underwent a total of 100 mannitol infusions. The effect of treatment was considered per sign and per symptom according to the Veldman et al. criteria for CRPS I. RESULTS: Overall improvement of CRPS I after mannitol treatment was successful in 24% after 1 week, and in 30% after 1 month. Mannitol treatment had some effect in patients with initially warm CRPS I in contrast to patients with cold CRPS I (OR=6.30 with CI [2.37-16.75]). Also patients with CRPS I at the upper extremity had more benefit than patients with CRPS I at the lower extremity (OR=3.26 with CI [1.34-7.93]). Poor results of mannitol treatment were associated with cold CRPS I (p<0.001), chronic CRPS I (p=0.04) and multiple mannitol treatments (p=0.04). CONCLUSION: Mannitol did not significantly contribute to the overall success of treatment in patients with CRPS I. Patients, presenting with acute, warm CRPS I in the upper extremity may have some benefit. PMID- 20018282 TI - Prepregnancy body mass index, gestational weight gain, and pregnancy outcomes in China. AB - OBJECTIVE: To look for associations between pregnancy outcomes and prepregnancy body mass index and gestational weight gain among Han women from Shenyang province, China. METHOD: A total of 2586 women were distributed across 4 prepregnancy categories according to the Chinese classification of body mass index, and to 4 end-of-pregnancy categories according to median weekly gestational weight gain. RESULTS: The risks for gestational hypertension, pre eclampsia, gestational diabetes, and preterm premature rupture of membranes were higher for those who were overweight or obese before becoming pregnant (P<0.05). Moreover, a gestational weight gain of 0.50 kg per week or greater was associated with a higher risk for gestational hypertension, preterm premature rupture of membranes, and fetal macrosomia (P<0.05). Women in the highest quartile for weight gain (>or=0.59 kg per week) were at higher risk for pre-eclampsia (P<0.05). CONCLUSION: A high prepregnancy body mass index and excessive gestational weight gain were associated with increased risks for adverse pregnancy outcomes. PMID- 20018283 TI - Clear detection of ADIPOQ locus as the major gene for plasma adiponectin: results of genome-wide association analyses including 4659 European individuals. AB - OBJECTIVE: Plasma adiponectin is strongly associated with various components of metabolic syndrome, type 2 diabetes and cardiovascular outcomes. Concentrations are highly heritable and differ between men and women. We therefore aimed to investigate the genetics of plasma adiponectin in men and women. METHODS: We combined genome-wide association scans of three population-based studies including 4659 persons. For the replication stage in 13795 subjects, we selected the 20 top signals of the combined analysis, as well as the 10 top signals with p values less than 1.0 x 10(-4) for each the men- and the women-specific analyses. We further selected 73 SNPs that were consistently associated with metabolic syndrome parameters in previous genome-wide association studies to check for their association with plasma adiponectin. RESULTS: The ADIPOQ locus showed genome-wide significant p-values in the combined (p=4.3 x 10(-24)) as well as in both women- and men-specific analyses (p=8.7 x 10(-17) and p=2.5 x 10(-11), respectively). None of the other 39 top signal SNPs showed evidence for association in the replication analysis. None of 73 SNPs from metabolic syndrome loci exhibited association with plasma adiponectin (p>0.01). CONCLUSIONS: We demonstrated the ADIPOQ gene as the only major gene for plasma adiponectin, which explains 6.7% of the phenotypic variance. We further found that neither this gene nor any of the metabolic syndrome loci explained the sex differences observed for plasma adiponectin. Larger studies are needed to identify more moderate genetic determinants of plasma adiponectin. PMID- 20018284 TI - Atorvastatin inhibits homocysteine-induced oxidative stress and apoptosis in endothelial progenitor cells involving Nox4 and p38MAPK. AB - Previous studies showed that homocysteine (Hcy) reduces endothelial progenitor cells (EPCs) numbers and impairs functional activity. Atorvastatin, HMG-CoA inhibition has been showed to have protective effects on EPCs. Recent studies have demonstrated that reduced EPCs numbers and activity are associated with EPCs apoptosis. However, the protective mechanisms of atorvastatin on HHcy-induced EPCs apoptosis remain to be determined. This study was designed to examine the effect of atorvastatin on homocysteine-induced reactive oxygen species (ROS) production and apoptosis in EPCs. EPCs were isolated from peripheral blood and characterized, then challenged with Hcy (50-500 micromol/L) in the presence or absence of atorvastatin (0.01-1 micromol/L) or various stress signaling inhibitors, including mevalonate (100 micromol/L), antioxidants N-acetyl cysteine (NAC, 10 micromol/L), the nicotinamide adenine dinucleotide phosphate (NADPH) oxidase inhibitor diphenylene iodonium (DPI 10 micromol/L), the eNOS inhibitor N(G)mono-methyl-l-arginine LNMA (1mmol/L), and the p38 mitogen-activated protein kinase (p38 MAPK) inhibitor SB203580 (10 micromol/L). Apoptosis was evaluated by FACS analysis and cell viability was determined by MTT assay. ROS were detected by 2',7'-dichlorodihydrofluorescein diacetate (H(2)DCFH-DA). NADPH oxidases were evaluated with lucigenin-enhanced chemiluminescence. Expression of Nox4 mRNA and p-p38MAPK protein was measured by RT-PCR and Western blot analysis, respectively. Our data revealed that atorvastatin significantly suppressed Hcy-induced ROS accumulation and EPCs apoptosis. Atorvastatin also antagonized homocysteine induced activation of NADPH oxidase and overexpression of Nox4 mRNA and p-p38MAPK protein. Similar effects occurred with EPCs transfected with Nox4 siRNA. These findings demonstrated that atorvastatin may inhibit Hcy-induced NADPH oxidase activation, ROS accumulation, and EPCs apoptosis through Nox4/p38MAPK dependent mechanisms, all of which may contribute to atorvastatin-induced beneficial effects on EPCs function. PMID- 20018285 TI - Pseudoxanthoma elasticum and familial hypercholesterolemia: a deleterious combination of cardiovascular risk factors. AB - BACKGROUND AND OBJECTIVE: Pseudoxanthoma Elasticum (PXE), an autosomal recessive disease due to mutations in ABCC6 gene, is characterised by fragmentation of elastic fibres with involvement of the cardiovascular system. We investigated a 60-year-old female with angina pectoris found to have PXE, associated with elevated plasma LDL-C suspected to be due to autosomal-co-dominant hypercholesterolemia. METHODS: ABCC6, LDLR, PCSK9 and exon 26 of APOB genes were re-sequenced. Cardiovascular involvement was assessed by coronary angiography, single-photon emission computed tomography (SPECT) and ultrasound examination. RESULTS AND CONCLUSIONS: The patient was a compound heterozygous for two ABCC6 mutations (p.S317R and p.R1141X) and heterozygous for a novel LDLR mutation (p.R574H). She had severe coronary stenosis and calcification of the arteries of the lower limbs. Treatment with ezetimibe/simvastatin 10/60mg/day, maintained over a 4.5-year period, reduced of LDL-C and the myocardial ischemic area. In PXE patients LDL-lowering treatment might contribute to delay macrovascular complications. PMID- 20018286 TI - Early vascular and metabolic effects of rosuvastatin compared with simvastatin in patients with type 2 diabetes. AB - OBJECTIVE: To compare the short-term effects of rosuvastatin and simvastatin on insulin-resistance and endothelial dysfunction in middle-aged patients with type 2 diabetes and mild untreated dyslipidemia. METHODS AND DESIGN: 29 Subjects randomly assigned to rosuvastatin 20mg/daily or simvastatin 20mg/daily for 4 weeks. Following data collected both pre- and post-treatment: fasting glucose, lipids, hs CRP, TNF-alpha, insulin sensitivity measured with euglycemic hyperinsulinemic clamp and flow-mediated dilation with brachial artery reactivity technique. RESULTS: Both treatments markedly reduced LDL cholesterol (p<0.001 for both). Insulin sensitivity did not change from relative baseline values in both groups, as well as fasting glucose and adiponectin. Simvastatin significantly improved flow-mediated dilation (p<0.01), to a greater extent than in patients taking rosuvastatin (p=0.09). We found no association between flow-mediated dilation improvement, LDL reduction and changes in hs CRP levels. CONCLUSION: In type 2 diabetic individuals rosuvastatin was less effective than simvastatin at improving endothelium-dependent vasodilation within one month, without affecting insulin-resistance, adiponectin levels and inflammation. PMID- 20018287 TI - The effect of positioning on the biomechanical performance of soft shell hip protectors. AB - Wearable hip protectors represent a promising strategy for reducing risk for hip fracture from a sideways fall. However, small changes in pad positioning may influence their protective benefit. Using a mechanical hip impact simulator, we investigated how three marketed soft shell hip protectors attenuate and redistribute the impact force applied to the hip, and how this depends on displacement from their intended position by 2.5 or 5 cm superiorly, posteriorly, inferiorly or anteriorly. For centrally-placed protectors, peak pressure was reduced 93% below the unpadded value by a 16 mm horseshoe-shaped protector, 93% by a 14 mm horseshoe protector, and 94% by a 16 mm continuous protector. In unpadded trials, 83% of the total force was applied to the skin overlying the proximal femur (danger zone). This was lowered to 19% by the centrally placed 16 mm horseshoe protector, to 34% by the 14 mm horseshoe, and to 40% by the 16 mm continuous protector. Corresponding reductions in peak force delivered to the femoral neck (relative to unpadded) were 45%, 38%, and 20%, respectively. The protective benefit of all three protectors decreased with pad displacement. For example, displacement of protectors by 5 cm anteriorly caused peak femoral neck force to increase 60% above centrally-placed values, and approach unpadded values. These results indicate that soft shell hip protectors provide substantial protective benefits, but decline in performance with small displacements from their intended position. Our findings confirm the need for correct and stable positioning of hip protectors in garment design. PMID- 20018288 TI - Mathematical modeling and simulation of seated stability. AB - Various methods have been used to quantify the kinematic variability or stability of the human spine. However, each of these methods evaluates dynamic behavior within the stable region of state space. In contrast, our goal was to determine the extent of the stable region. A 2D mathematical model was developed for a human sitting on an unstable seat apparatus (i.e., the "wobble chair"). Forward dynamic simulations were used to compute trajectories based on the initial state. From these trajectories, a scalar field of trajectory divergence was calculated, specifically a finite time Lyapunov exponent (FTLE) field. Theoretically, ridges of local maxima within this field are expected to partition the state space into regions of qualitatively different behavior. We found that ridges formed at the boundary between regions of stability and failure (i.e., falling). The location of the basin of stability found using the FTLE field matched well with the basin of stability determined by an alternative method. In addition, an equilibrium manifold was found, which describes a set of equilibrium configurations that act as a low dimensional attractor in the controlled system. These simulations are a first step in developing a method to locate state space boundaries for torso stability. Identifying these boundaries may provide a framework for assessing factors that contribute to health risks associated with spinal injury and poor balance recovery (e.g., age, fatigue, load/weight, and distribution). Furthermore, an approach is presented that can be adapted to find state space boundaries in other biomechanical applications. PMID- 20018289 TI - Improvement of diamond-like carbon electrochemical corrosion resistance by addition of nanocrystalline diamond. AB - Nanocrystalline diamond (NCD) particles were incorporated into diamond-like carbon (DLC) films in order to investigate NCD-DLC electrochemical corrosion resistance. The films were grown over 304 stainless steel using plasma-enhanced chemical vapor deposition. NCD particles were incorporated into DLC during the deposition process. The investigation of NCD-DLC electrochemical corrosion behavior was performed using potentiodynamic polarization against NaCl. NCD-DLC films presented more negative corrosion potential and lower anodic and cathodic current densities. The electrochemical analysis indicated that NCD-DLC films present superior impedance and polarization resistance compared to the pure DLC, which indicate that they are promising corrosion protective coatings in aggressive solutions. PMID- 20018290 TI - Electrokinetic properties of hydroxyapatite under flotation conditions. AB - The effect of calcite supernatant, calcium, and carbonate ions on the hydroxyapatite (HA) zeta potential without and in the presence of sodium oleate (1x10(-4) mol L(-1)) was examined within the pH range from 4 to 12. The interpretation of results was based on the HA surface and oleate solution chemistry, and on some floatability tests. HA, with different positive and negative surface sites formed depending on its solubility and pH, had a negative zeta potential over the whole pH range. This mineral is not naturally floatable (flotation recovery, 5%9.3. The HA surface is less negatively charged in calcite supernatant than in water from pH 6.6 to 9.2 due to the adsorption on HA negative surface active centers ([triple bond]HPO(4)(-) and [triple bond]PO(4)(2 )) of the Ca(2+), and CaOH(+) ions (present in the calcite supernatant), producing more surface sites [triple bond]HPO(4)Ca(+), [triple bond]PO(4)Ca, [triple bond]HPO(4)CaOH, and [triple bond]PO(4)(-) CaOH, and new centers [triple bond]HPO(4)CaHCO(3) and [triple bond]PO(4)(-) CaHCO(3). In the presence of 1x10( 3) mol L(-1) CaCl(2), the HA sample has positive zeta potential, the same as calcite from the same deposit, up to IEP at pH 11.25. Carbonate ions (1x10(-3) mol L(-1) Na(2)CO(3)) do not affect the HA zeta potential. However, a possible process can be the ion-exchange reaction between bicarbonate (or carbonate) and some anion from the surface sites formed on HA. The obtained values of the HA zeta potential with the collector (1x10(-4) mol L(-1) Na-oleate) added into hydroxyapatite/calcite supernatant suspensions corroborate the weak chemisorption of Ol(-) and H(Ol)(2)(-). The likely processes in this system also are the ion exchange reactions on [triple bond]HPO(4)CaOH and [triple bond]PO(4)(-) CaOH, [triple bond]HPO(4)CaHCO(3) and [triple bond]PO(4)(-) CaHCO(3) between oleate ion and surface hydroxyl and bicarbonate ions, surface and bulk precipitations of calcium oleate, Ca(Ol)(2), and the surface and bulk precipitations of Ca[H(Ol)(2)(-)](2) over the pH range from 7 to 9. Calcite supernatant does not influence natural floatability of the mineral. However, calcite supernatant depresses the hydroxyapatite flotation in the presence of 1x10(-4) mol L(-1) Na oleate (pH 9, R approximately 50%), a likely result of the weak chemisorption due to the steric effect of heterogeneous HA surface formed in calcite supernatant, Ca(Ol)(2) and Ca[H(Ol)(2)(-)](2) surface and bulk precipitations. PMID- 20018291 TI - A disjoining pressure study of foam films stabilized by mixtures of a nonionic (C12DMPO) and an ionic surfactant (C12TAB). AB - This work is a continuation of a previous study [Langmuir 23 (2007) 5315-5323] of aqueous foam films stabilized by surfactant mixtures consisting of a cationic and a nonionic surfactant. In both studies a thin film pressure balance was used to investigate the disjoining pressure Pi as a function of the film thickness h. With the resulting Pi-h curves the surface charge densities q(0) of the foam films were determined by fitting them with the Derjaguin-Landau-Verwey-Overbeek (DLVO) theory. Our previous study demonstrated that mixing the nonionic surfactant beta-dodecylmaltoside (beta-C(12)G(2)) and the cationic dodecyl trimethylammonium bromide (C(12)TAB) allows one to tune q(0) and thus the type of foam film. In the present study the nonionic surfactant dodecyldimethyl phosphineoxide (C(12)DMPO) instead of beta-C(12)G(2) was used and C(12)DMPO/C(12)TAB mixtures in a concentration range from 0.1 to 2.0 cmc were studied at mixing ratios of C(12)DMPO:C(12)TAB = 1:0, 50:1, 1:1, 1:50, 0:1. The results are compared with those of the beta-C(12)G(2)/C(12)TAB mixture and are discussed in terms of surface charge densities. PMID- 20018292 TI - An XPS study for mechanisms of arsenate adsorption onto a magnetite-doped activated carbon fiber. AB - The surface and bulk structures of a newly developed carbon-based iron-containing adsorbent for As(V) adsorption were investigated by using X-ray diffraction (XRD), field emission scanning electronic microscopy (FESEM), and X-ray photoelectron spectroscopy (XPS). XRD patterns of the adsorbents indicated that the modified activated carbon fiber (MACF) was a simple mixture of the raw activated carbon fiber (RACF) and magnetite. After modification, a porous film was formed on the surface of the MACF with nano-sized magnetite on it. The As(V) uptake on the MACF was highly pH dependent and was facilitated in acidic solutions. XPS studies demonstrated that the surface oxygen-containing functional groups were involved in the adsorption and that magnetite played a key role in As(V) uptake. The dominance of HAsO(4)(2-) in surface complexes and the pH effect on As(V) uptake demonstrated that the monoprotonated bidentate complexes were dominant on the surface of the MACF. No reduction of As(V) was observed on the surface of the ACFs. PMID- 20018293 TI - Phase transition behaviors of poly(N-isopropylacrylamide) microgels induced by tannic acid. AB - Effects of tannic acid (TA) on the phase transition behaviors of monodisperse poly(N-isopropylacrylamide) (PNIPAM) microgels in aqueous solutions are systematically investigated. Monodisperse PNIPAM microgels are prepared through a microfluidic approach. The isothermal phase transition behaviors of PNIPAM microgels induced by TA in aqueous solutions are studied at 25 degrees C. The results show that there exist significant isothermal phase transition phenomena for PNIPAM microgels induced by TA, and the dynamic phase transition rate and the equilibrium shrinking degree of PNIPAM microgels in TA solutions are dependent on the TA concentration. The higher the TA concentration, the faster the isothermal shrinking rate of PNIPAM microgels induced by TA. There exists a critical TA concentration for the isothermal phase transition behavior, below which the equilibrium shrinking degree of PNIPAM microgels becomes larger with increasing the TA concentration, while above which the equilibrium shrinking degrees of PNIPAM microgels remain almost the same. The adsorption of TA molecules to PNIPAM polymer networks leads to slight shift of the low critical solution temperature to higher temperatures when the TA concentration in solution is lower than 10(-5) mol/L. PMID- 20018294 TI - Optically switchable organic hollow nanocapsules. AB - Hollow nanocapsules with both photoswitchable-fluorescent and reversible photochromic properties are synthesized via a one-pot non-templating microemulsion copolymerization using methyl methacrylate and methacrylated spiropyran as co-monomers. The strong photoswitchable fluorescence of the nanocapsules are switched between "on" and "off" by alternating irradiation of ultraviolet and visible light, which causes the reversible photoisomerization between spiropyran and merocyanine in the nanocapsules. The distribution of spiropyran/merocyanine in a nanocapsule is mostly incorporated inside the wall of the nanocapsule, with only about 17.7% on the surface of the wall. This confinement is the reason of the unusually strong fluorescence of the merocyanine form yielded by the UV radiation. For the same reason, the photochemical stability of the chromophores is increased compared to those in the solution of water/DMF mixture. PMID- 20018295 TI - Consideration of polydispersity in the evaluation of the dynamic mobility of concentrated suspensions. AB - Many practical uses of electroacoustic methods for the characterization of disperse systems involve concentrated and/or polydisperse suspensions. While the effects of particle concentration have been well described experimentally and theoretically, similar studies considering a wide size distribution of the dispersed particles are lacking. This is not a minor point, as these methods are based on the action of alternating fields (either electric or acoustic) on the systems and the characteristic frequencies and amplitudes are largely determined by the particle geometry. In this work, we first evaluate the effect, on the dynamic (or ac) mobility, of changing the size distribution in the suspension. It is found that the inertia (also called hydrodynamic) relaxation of the mobility is shifted toward lower frequencies, and that the overall mobility spectrum is smoothed when the size polydispersity of the suspension increases. The results theoretically obtained are subsequently used for fitting experimental mobility data corresponding to two alumina samples, in a wide range of particle concentrations and ionic strengths. It is demonstrated that a complete model accounting for polydispersity leads to a better description of the results; very significantly, this can be done by using the zeta potential as the only fitting parameter, and forcing this parameter to be determined only by the ionic strength, and not by the volume fraction. PMID- 20018296 TI - Investigating the predictive roles of working memory and IQ in academic attainment. AB - There is growing evidence for the relationship between working memory and academic attainment. The aim of the current study was to investigate whether working memory is simply a proxy for IQ or whether there is a unique contribution to learning outcomes. The findings indicate that children's working memory skills at 5 years of age were the best predictor of literacy and numeracy 6 years later. IQ, in contrast, accounted for a smaller portion of unique variance to these learning outcomes. The results demonstrate that working memory is not a proxy for IQ but rather represents a dissociable cognitive skill with unique links to academic attainment. Critically, we find that working memory at the start of formal education is a more powerful predictor of subsequent academic success than IQ. This result has important implications for education, particularly with respect to intervention. PMID- 20018297 TI - Significance of HER2 protein examination in ductal carcinoma in situ. AB - BACKGROUND: HER2 expression is routinely checked in ductal carcinoma in situ, as in invasive ductal carcinoma. However, the effect of HER2 status in ductal carcinoma in situ on the development of malignancy and the significance of overexpression of HER2 are still not clear. MATERIALS AND METHODS: We experienced 103 cases that were diagnosed as pure ductal carcinoma in situ from operative specimens in the 2-y period from 2006 to 2007. We examined their HER2 status and other markers. We added 38 cases of ductal carcinoma in situ with small invasive disease 5mm or less in diameter as subjects. We also examined how accurately HER2 status in biopsy specimens predicted the existence of an invasive component. RESULTS: In pure ductal carcinoma in situ, tumors that were comedo type, high grade, or ER negative showed a high frequency of HER2 overexpression. In cases with small invasion, HER2 expression was higher than that in pure ductal carcinoma in situ. Among cases that were diagnosed as ductal carcinoma in situ by biopsy, 28% had invasive disease in operative specimens. In tumors that were palpable, large, or expressed HER2 3+ in biopsy samples, invasive disease was frequently observed in operative specimens. CONCLUSIONS: Overexpression of HER2 in ductal carcinoma in situ might not always be necessary for progression to invasive ductal carcinoma. To clarify the significance of HER2 examination in DCIS, further investigations of the potential for invasive ductal carcinoma and the prognosis are still needed. PMID- 20018298 TI - Enhanced expression and clinical significance of chemokine receptor CXCR2 in hepatocellular carcinoma. AB - BACKGROUND: An ELR+ CXC chemokine receptor, CXCR2, was recently reported to be involved in tumorigenesis and development. However, the role of CXCR2 in hepatocellular carcinoma (HCC) is poorly understood. In this study, we aimed to investigate the association between CXCR2 expression and the biocharacteristics of HCC, and determine whether the expression of CXCR2 was related to the tumorigenesis and progression. METHODS: Forty-two patients who underwent hepatic resection and were diagnosed as HCC by histologic examination were included. HCC and corresponding adjacent tissues (distance from the tumor border exceeding 2 cm) were obtained. Twenty-three samples of normal liver tissue were acquired surgically from the patients who had received an operation due to liver trauma. CXCR2 mRNA and protein expressions were examined using semiquantitative reverse transcription-PCR and Western blot. RESULTS: Expression levels of CXCR2 were significantly increased in HCC compared with adjacent and normal liver tissues (P < 0.05). The results showed that CXCR2 mRNA and protein expression levels were not related to age, gender, AFP levels, tumor capsule, or tumor size. Also, there was no relationship between CXCR2 mRNA expression and TNM staging. The expression levels of CXCR2 mRNA and protein were correlated with intrahepatic metastasis (P < 0.05), portal cancer embolus (P < 0.05), and low differentiation (P < 0.05). Furthermore, the protein level of CXCR2 was relevant to TNM staging. The protein level of CXCR2 in stage III-IV was remarkably higher than in stage I-II (P < 0.05). CONCLUSION: Our data revealed that CXCR2 was able to promote invasion and metastasis of HCC. It may be a useful marker for judging biocharacteristics and prognosis of HCC. PMID- 20018299 TI - WITHDRAWN: Influences of Perfusion Pressure and Pulsatile Flow on Cerebral Flow Reserve and Oxygenation in the Isolated Perfused Swine Brain under Normothermia. AB - This article has been removed at the request of the Editor-in-Chief. Please see Elsevier Policy on Article Withdrawal: (http://www.elsevier.com/locate/withdrawalpolicy). PMID- 20018300 TI - Pretreatment with dexmedetomidine or thiopental decreases myoclonus after etomidate: a randomized, double-blind controlled trial. AB - BACKGROUND: Myoclonic movements are common problems during induction of anesthesia with etomidate. The aim of this study was to compare the effect of pretreatment with dexmedetomidine (0.5 microg/kg) and thiopental (1 mg/kg) on the incidence of etomidate-induced myoclonus and postoperative pain. MATERIALS AND METHODS: A prospective double-blind study was conducted at a university hospital. Ninety patients (ASA physical status I-II) were randomly assigned to one of three groups: patients were pretreated with either dexmedetomidine (0.5 microg/kg), thiopental (1 mg/kg), or saline before induction of anesthesia with etomidate. One minute after the injection of study drugs, etomidate, 0.3 mg/kg was given. Myoclonus was assessed on a scale of 0 to 3. Recovery time, postoperative pain score, and hemodynamic variables were recorded during the intraoperative and postoperative period. Headache, nausea, vomiting, and coughing were noted during the study. RESULTS: The incidence and the intensity of myoclonus was significantly lower in the dexmedetomidine and thiopental groups (34%, 36%) than in the control group (64%) (P<0.05). The postoperative pain score at 30 min in the thiopental group was significantly higher than in the dexmedetomidine and control groups (63%) (P<0.05). CONCLUSIONS: We concluded that pretreatment with dexmedetomidine or thiopental is effective in reducing the incidence and severity of etomidate-induced myoclonic muscle movements and pretreatment with thiopental increases the postoperative pain. PMID- 20018301 TI - Simvastatin induces heme oxygenase-1 expression but fails to reduce inflammation in the capsule surrounding a silicone shell implant in rats. AB - BACKGROUND: Statins are lipid-lowering agents that are widely used in medical practice. Some studies have demonstrated that statins reduce proinflammatory signaling and have antioxidant and anti-inflammatory effects. The inducible heme oxygenase-1 (HO-1) catalyzes the stepwise degradation of heme to produce equimolar quantities of biliverdin, iron, and carbon monoxide; it has also been suggested to promote the important cholesterol-independent cytoprotective action of statins against oxidative insults. However, overexpression of HO-1 may have dual effects under oxidative stress, and the overaccumulation of ferric iron from heme may result in detrimental rather than cytoprotective effects. This study was designed to investigate the effect of a specific statin, simvastatin, and the role of HO-1 on the inflammatory status of the capsule surrounding a silicone shell implant in rats. METHODS: Sprague-Dawley rats were randomly assigned to different groups according to the indicated implantation time (1 , 2 wk, 1 , 2 , and 3 mo postimplant) and the presence or absence of additional treatment (2 mg/kg oral simvastatin daily and/or 2 mg/kg HO inhibitor zinc protoporphyrin (ZnPP) by intraperitoneal injection daily). The formed de novo capsular tissue around the silicone implant was dissected away at the indicated time of sacrifice for experiments regarding the HO-1 expression and the inflammatory status. RESULTS: We found a biphasic pattern of inflammation of the capsule with increased production of TNF-alpha, IL-1beta, and COX-2, and activation of NF kappaB at 1 wk and again at 1 mo postimplantation. Numerous CD68+monocytes or macrophages were diffusely distributed in the capsule in the early stage but not in the late stage postimplantation. In addition, the implantation procedure induced HO-1 expression, which was significantly up-regulated in the first 2 wk but decreased after 1 mo. Although daily feeding of the rats with simvastatin at 2 mg/kg induced HO-1 expression in the capsule throughout the experimental period, simvastatin treatment failed to reduce the production of TNF-alpha, IL 1beta, and COX-2, or limit NF-kappaB activation in the early or late stages after implantation. Additionally, simvastatin treatment did not decrease the amount of CD68+cells distributed in the capsule at 1 wk postimplantation. HO-1 up regulation by simvastatin treatment or down-regulation by additional injection of ZnPP did not demonstrate significant correlation with the inflammatory status of the capsule. CONCLUSION: Simvastatin failed to reduce inflammation of the capsule surrounding a silicone shell implant. Although simvastatin induced HO-1 expression throughout the experimental time, up-regulated HO-1 expression could not be attributed to the inflammatory status of the capsule. PMID- 20018302 TI - Curcumin provides neuroprotection after spinal cord injury. AB - BACKGROUND: Traumatic spinal cord injury (SCI) is a major cause of long-term disability. However, therapeutic agents targeting SCI are sorely lacking. The aim of this study was to investigate whether curcumin has neuroprotective effects after SCI in rats. MATERIALS AND METHODS: Studies were performed in 39 male Sprague-Dawley rats after spinal cord hemisection. The animals were randomly divided into three groups: sham, vehicle, and curcumin. The Basso, Beattie, and Bresnahan (BBB) scale was used to evaluate functional outcome. Specimens were tested for histologic, terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick-end labeling (TUNEL), and immunohistochemical staining. Primary cultured astrocytes were used to test the inhibitory effect of curcumin on glial reactivation. RESULTS: The BBB scores for the affected hindlimb after hemisection were significantly improved in the curcumin-treated group compared with the vehicle group (on d 3 and 7; P < 0.001). Immunohistochemistry of NeuN revealed remarkable neuronal loss in the vehicle group after hemisection. In comparison, curcumin significantly protected neurons after SCI (curcumin compared with vehicle; P < 0.001). Furthermore, curcumin significantly attenuated apoptosis after SCI (curcumin compared with vehicle; P < 0.001). RT-PCR demonstrated that the expression of glial fibrillary acidic protein (GFAP) was significantly inhibited by curcumin. CONCLUSIONS: Curcumin inhibited apoptosis and neuron loss, quenched astrocyte activation, and significantly improved neurologic deficit 7 d after spinal cord hemisection. By down-regulating GFAP expression, curcumin seems to attenuate astrocyte reactivation, which may be beneficial for neuronal survival. This is the first report demonstrating the successful treatment of SCI by curcumin. PMID- 20018303 TI - Expression of tissue factor pathway inhibitor 2 in human pancreatic carcinoma and its effect on tumor growth, invasion, and migration in vitro and in vivo. AB - Tissue factor pathway inhibitor 2 (TFPI-2), also known as placental protein and matrix-associated serine protease inhibitor, plays an important role in angiogenesis, intravascular fibrinolysis, wound healing, tumor invasion, metastasis by plasmin, and trypsin mediated activation of zymogen matrix metalloproteinases. To detect whether TFPI-2 can be expressed in human pancreatic carcinoma samples and to investigate its role in the growth, invasion, and metastasis of pancreatic carcinoma cell in vitro and in vivo, we collected eight normal pancreatic tissue samples and 50 pancreatic carcinoma samples, and stably transfected the human pancreatic carcinoma cell line Panc-1 with a vector capable of expressing TFPI-2 gene. RT-PCR and Western blot analysis revealed that the levels of TFPI-2 expression were markedly lower in pancreatic carcinoma samples compared with normal pancreas samples. The level of TFPI-2 protein was significantly higher in cells transfected with TFPI-2 gene than that in the untransfected cells. The results of MTT assay showed that TFPI-2 inhibited Panc-1 cells growth in vitro. The invasive capacity of the cells transfected with TFPI-2 gene was also markedly less than that of untransfected cells in vitro as determined by the Matrigel invasion/migration assay. Moreover, TFPI-2 inhibited tumor growth, invasion, and metastasis in vivo in an orthotopic pancreatic carcinoma model. Our findings suggest that TFPI-2 plays a significant role in the growth, invasion, and metastasis of pancreatic carcinoma cell in vitro and in vivo, and has potential in anticancer therapy. PMID- 20018304 TI - Diverse effects of FK506 on the apoptosis of hepatocytes and infiltrating lymphocytes in an allografted rat liver. AB - BACKGROUND: The current study investigated whether FK506 (FK) regulates the apoptotic systems in allografted rat liver and the contribution of Fas/Fas-ligand system and Bcl-2 family during acute rejection. MATERIALS AND METHODS: The recipients were divided into three groups, the allo, the allo-FK, and the syn group. Rats were euthanized 1, 3, 5, and 7 d after OLT. Apoptotic activity was explored using terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL) assay. The expression of Fas/Fas-ligand and Bcl-2/Bax in the grafted livers was investigated by Western blotting and immunohistochemistry. RESULTS: The apoptotic index (AI) of hepatocytes in the allo-FK group was less than that in the allo group. Fas in the allo group was more intense than that in the allo-FK group in the periportal areas on day 1 and 3, while Bcl-2 in the allo group was less intense than that in the allo-FK group in the pericaval areas at all time-points after OLT. CONCLUSION: FK provides beneficial antiapoptotic effects on hepatocytes in the grafted rat livers through both the down-regulation of Fas expression in the periportal areas and the up-regulation of Bcl-2 expression in the pericaval areas. PMID- 20018305 TI - Caseous calcification of the anterior mitral valve annulus presenting as intracardiac mass. PMID- 20018306 TI - Changes in cardiac and cognitive function and self-reported outcomes at one year after coronary artery bypass grafting. AB - OBJECTIVE: Although health status after coronary artery bypass grafting improves at the group level, individual outcomes demonstrate variation. We aimed to evaluate relative importances of changes in cardiac and cognitive function and symptom status regarding physical and mental health at 1 year after coronary artery bypass grafting. METHODS: Outcomes in multivariable regression analysis (n = 86) were self-reported physical and mental health (Medical Outcomes Study 36 Item Short Form) at 12 months' follow-up, adjusting for baseline. Independent variables were change in exercise capacity (staged ergometer protocol), cognitive function (neurocognitive test battery), and self-reported improvement of angina. Graft patency was evaluated by angiography (82/86 patients). RESULTS: After surgery, health status was comparable to the age- and sex-matched population norm. Improvement of angina was associated with gain in physical health in the range of 0.5 SD (beta = 0.23, P = .012). Change in observed physical exercise capacity (mean +/- SD 199 +/- 426 W x min) accounted for 9% of variance in physical health (beta = 0.42, P = .001). Positive change in cognitive function was significantly related to mental (beta = 0.52, P = .007) but not physical health. Graft occlusion (22/82 patients) was not significantly related to health status. CONCLUSIONS: Individual variation in objective functional performance and symptom relief is significant for self-reported health status, beyond the predictive ability of preoperative health status, sex, and graft patency. Awareness of the extent and impact of outcome variation is important when counseling patients before and after coronary artery bypass grafting. PMID- 20018307 TI - Reduction in incidence of deep sternal wound infections: random or real? AB - OBJECTIVE: Comorbidities predisposing cardiac surgical patients toward deep sternal wound infection, such as diabetes and obesity, are rising in the United States. Longitudinal analysis of risk factors, morbidity, and mortality was performed to assessed effects of these health trends on deep sternal wound infection rates. METHODS: In this retrospective analysis of all median sternotomies performed at a single institution from 1991 through 2006, demographic and surgical characteristics were identified from a prospective database. The cohort was separated into periods from 1992 through 2001 and 2002 through 2006 to identify longitudinal trends in risk factors for deep sternal wound infection. Univariate and matched multivariable analyses were performed. RESULTS: Overall, study population had increased comorbidities associated with deep sternal wound infection such as obesity, diabetes, and advanced age. Deep sternal wound infections were treated in 285 of 21,000 sternotomies performed during study period (1.35%). Deep sternal wound infection rates decreased from 1.57% to 0.88% in last 5 years. Rate of deep sternal wound infection was reduced among patients with diabetes from 3.2% to 1.0%. Multivariable analysis showed diabetes and smoking to be eliminated as risk factors in last 5 years. Prolonged bypass time was the only variable independently associated with deep sternal wound infection for the entire period. Thirty-day and 1-year mortalities for deep sternal wound infection did not change significantly. CONCLUSIONS: Analysis of a large series of cardiac surgical patients demonstrates significant reduction in deep sternal wound infection incidence in 15 years. Introduction of perioperative intravenous insulin may explain some observed risk reduction. Efforts should focus on prevention, because mortality remains elevated. PMID- 20018308 TI - Does neoadjuvant hormonal therapy improve urinary function when given to men with large prostates undergoing prostate brachytherapy? AB - PURPOSE: We evaluated the effect of neoadjuvant hormonal therapy on urinary function in men with a prostate volume of 50 cc or greater undergoing prostate brachytherapy. MATERIALS AND METHODS: A total of 395 men with 50 cc or greater glands were treated with 3 months of neoadjuvant hormonal therapy (204) or implantation alone (191). Urinary function was assessed by the International Prostate Symptom Score, the urinary retention incidence and subsequent transurethral prostate resection. RESULTS: Median patient age was 67 years and median followup was 6 years. Mean prostate volume in neoadjuvant hormonal therapy cases was 72.9 cc, which decreased to 54.3 cc after 3 months (p <0.001). Mean prostate volume in cases without hormonal therapy was 60.6 cc (p <0.001). Urinary retention occurred in 16 of 191 men (8.4%) without vs 25 of 204 (12.3%) with hormonal therapy (p = 0.207). The median duration of urinary retention was 42 days (range 2 to 243). There were no significant associations of urinary retention with prostate size, prostate or urethral dose, or pre-implantation International Prostate Symptom Score. Of patients without hormonal therapy retention occurred in 3 of 12 (25%) with a pre-implantation International Prostate Symptom Score of 15 or greater and in 13 of 168 (7.7%) with a score of less than 15 (OR 4.0, 95% CI 1-16, p = 0.04). In contrast, there was no difference in the retention rate in patients with hormonal therapy with an initial score of 15 or greater vs less than 15 (2 of 25 or 8% vs 11 of 102 or 10.8%, p = 0.614). Transurethral prostate resection was done in 11 of 191 men (5.8%) without vs 12 of 204 (5.9%) with hormonal therapy (p = 0.958). There was no difference in biochemical failure in the 2 groups. CONCLUSIONS: Neoadjuvant hormonal therapy has its greatest benefit in patients receiving brachytherapy who have a large prostate and an International Prostate Symptom Score of 15 or greater. PMID- 20018309 TI - Editorial comment. PMID- 20018310 TI - Editorial comment. PMID- 20018311 TI - Increased sperm DNA fragmentation in patients with vasectomy reversal has no prognostic value for pregnancy rate. AB - PURPOSE: We evaluated sperm DNA fragmentation in patients with vasectomy reversal and its prognostic value to determine spontaneous and assisted reproductive technique pregnancy rates. MATERIALS AND METHODS: We prospectively assessed DNA fragmentation with the sperm chromatin structure assay in postoperative semen samples of 70 patients with vasectomy reversal. At a median +/- SD followup of 4.3 +/- 0.5 years pregnancy rates were recorded. RESULTS: DNA fragmentation in patients with vasectomy reversal was significantly increased vs that in proven fertile controls (30.2% +/- 20.1% vs 15.3% +/- 5.4%, p <0.001). Significant negative correlations were found between DNA fragmentation index and total sperm count, progressive motility, total number of progressive sperm, normal morphology and sperm vitality (-0.325 30 kg/m2, soft gland texture, and main pancreatic duct size < or =3 mm. Predictive factors for DGE included 2 factors: not using a surgical microscope, and simultaneous PAF. CONCLUSION: A web-based calculator was developed to promote homogeneity of method for grading of PAF and DGE after PD. Anyone with access to the web can now compare their results to the current study. PMID- 20018336 TI - Characterisation of virus transport and attenuation in epikarst using short pulse and prolonged injection multi-tracer testing. AB - Attenuation processes controlling virus fate and transport in the vadose zone of karstified systems can strongly influence groundwater quality. This research compares the breakthrough of two bacteriophage tracers (H40/1 and T7), with contrasting properties, at subsurface monitoring points following application onto an overlying composite sequence of thin organic soil and weathered limestone (epikarst). Short pulse multi-tracer test results revealed that T7 (Source concentration, Co=1.8x10(6)pfu/mL) and H40/1 (Co=5.9x10(6)pfu/mL) could reach sampling points 10 m below ground less than 30 min after tracer application. Contrasting deposition rates, determined from simulated tracer responses, reflected the potential of the ground to differentially attenuate viruses. Prolonged application of both T7 (Co=2.3x10(4)pfu/mL) and H40/1 (Co=1.3x10(5)pfu/mL) over a five hour period during a subsequent test, in which ionic strength levels observed at monitoring points rose consistently, corresponded to a rapid rise in T7 levels, followed by a gradual decline before the end of tracer injection; this reflected reaction-limited deposition in the system. T7's response contrasted with that of H40/1, whose concentration remained constant over a three hour period before declining dramatically prior to the end of tracer injection. Subsequent application of lower ionic strength tracer-free flush water generated a rapid rise in H40/1 levels and a more gradual release of T7. Results highlight the benefits of employing prolonged injection multi-tracer tests for identifying processes not apparent from conventional short pulse tests. Study findings demonstrate that despite rapid transport rates, the epikarst is capable of physicochemical filtration of viruses and their remobilization, depending on virus type and hydrochemical conditions. PMID- 20018338 TI - Basic oxygen furnace slag as a treatment material for pathogens: contribution of inactivation and attachment in virus attenuation. AB - Basic oxygen furnace (BOF) slag media were studied as a potential treatment material in on-site sanitation systems. Batch and column studies were conducted to evaluate attenuation of the bacteriophage PR772 and 0.190 microm diameter microspheres by BOF media, and to delineate the relative contributions of two principle processes of virus attenuation: inactivation and attachment. In the batch studies, conducted at 4 degrees C, substantial inactivation of PR772 did not occur in the pH 7.6 and 9.5 suspensions. At pH 11.4, bimodal inactivation of PR772 was observed, at an initial rate of 2.1 log C/C(0) day(-1) for the first two days, followed by a much slower rate of 0.124 log C/C(0) day(-1) over the following 10 days. Two column studies were conducted at 4 degrees C at a flow rate of 1 pore volume day(-1) using two slag sources (Stelco, Ontario; Tubarao, Brazil) combined with sand and pea gravel. In both column experiments, the effluent microsphere concentration approached input concentrations over time (reductions of 0.1-0.2 log C/C(0)), suggesting attachment processes for microspheres were negligible. Removal of PR772 virus was more pronounced both during the early stages of the experiments, but also after longer transport times (0.5-1.0 log C/C(0)). PR772 reduction appeared to be primarily as a result of virus inactivation in response to the elevated pH conditions generated by the BOF mixture (10.6-11.4). On-site sanitation systems using BOF media should be designed to maintain sufficient contact time between the BOF media and the wastewater to allow sufficient residence time of pathogens at elevated pH conditions. PMID- 20018337 TI - Assessment of natural fluorescence as a tracer of diffuse agricultural pollution from slurry spreading on intensely-farmed grasslands. AB - The value of natural fluorescence in tracing diffuse pollution, in liquid phase, following slurry application to land was assessed by field experiment using twelve one hectare lysimeters on a heavy clay soil in Devon, UK, during autumn 2007. A strong linear relationship was found between natural fluorescence intensity and slurry concentration. The ratio of indices of tryptophan-like and fulvic/humic-like fluorescence (TI:FI) varied between 2 and 5 for a range of slurries sampled from Devon farms and allowed slurry to be distinguished from uncontaminated drainage waters (TI:FI<1). Incidental losses of slurry, indicated by significantly enhanced TI:FI ratios, high TI and high ammonium levels, occurred via the drain flow pathway of the drained lysimeters during the first small event following slurry-spreading. The maximum estimated loss from a single lysimeter was 2-8kg or 0.004-0.016% of the applied slurry. In the second larger storm event, some five weeks later, significantly enhanced TI:FI ratios in the drain flows were not associated with high TI but with high nitrate levels and, compared to the earlier storm, an increase in the humification index. This implies the loss of slurry decomposition products during this event but further work is needed to validate this. There was no significant enhancement of TI:FI in the surface/throughflow pathways of the drained or undrained lysimeters in either of the events. The observed change over a period of weeks in the strength and nature of the fluorescence signal from spread slurry restricts quantification of slurry losses to those immediately after slurry spreading. Nonetheless, this study demonstrates the utility of fluorescence as an indicator of slurry in drainage waters and the importance of field drains in diffuse agricultural pollution. PMID- 20018339 TI - Electronic circular dichroism of monomethyl [16O,17O,18O]-phosphate and [16O,17O,18O]-thiophosphate revisited. AB - Phosphoryl-transfer reactions have long been of interest due to their importance in maintaining numerous cellular functions. A phosphoryl-transfer reaction results in two possible stereochemical outcomes: either retention or inversion of configuration at the transferred phosphorus atom. When the product is phosphate, isotopically-labeled [16O,17O,18O]-phosphate derivatives can be used to distinguish these outcomes; one oxygen must be replaced by sulfur or esterified to achieve isotopic chirality. Conventionally, stereochemical analysis of isotopically chiral phosphate has been based on 31P NMR spectroscopy and involves complex chemical or enzymatic transformations. An attractive alternative would be direct determination of the enantiomeric excess using chiroptical spectroscopy. (S)-Methyl-[16O,17O,18O]-phosphate (MePi*), 7 and enantiomeric [16O,17O,18O] thiophosphate (TPi*), 10, were previously reported to exhibit weak electronic circular dichroism (ECD), although with 10 the result was considered to be uncertain. We have now re-examined the possibility that excesses of 7 and 10 enantiomers can be detected by ECD spectrometry, using both experimental and theoretical approaches. 7 and both the (R) and (S) enantiomers of 10 (10a,10b) were synthesized by the 'Oxford route' and characterized by 1H, 31P and 17O NMR, and by MS analysis. Weak ECD could be found for 7, with suboptimal S/N. No significant ECD could be detected for the 10 enantiomers. Time-dependent DFT (TDDFT) calculations of the electronic excitation energies and rotational strengths of the same three enantiomers were carried out using the functional B3LYP and the basis set 6-311G**. The isotopically-perturbed geometries were predicted using the anharmonic vibrational frequency calculational code in GAUSSIAN 03. In the case of 10, calculations were also carried out for the hexahydrated complex to investigate the influence of the aqueous solvent. The predicted excitation wavelengths are greater than the observed wavelengths, a not unusual result of TDDFT calculations. The predicted anisotropy ratios are 2.9 x 10(-5) for 7, -5.3 x 10(-6) for 10a/b, and 1.7 x 10(-6) for 10a/b.(H2O)6. For 7 the predicted anisotropy ratio approximates that observed in this work, 4.5 x 10( 5) at 208 nm. For 10a/b, the upper limits of the experimental anisotropy ratios (<5 x 10(-6) at 225 nm, pH 9; <5 x 10(-6) at 236 nm, pH 12) are comparable to the predicted magnitude of the value for 10a/b. The lower predicted value for 10a/b.(H2O)6 suggests that the aqueous environment affects the ECD significantly. Altogether, the TDDFT calculations together with a stereochemical analysis based on NMR and the MS data support the conclusion that the experimental ECD results for MePi* and TPi* may be reliable in order of magnitude. PMID- 20018340 TI - Synthesis of 4,6-dideoxy-3-fluoro-2-keto-beta-D-glucopyranosyl analogues of 5 fluorouracil, N6-benzoyl adenine, uracil, thymine, N4-benzoyl cytosine and evaluation of their antitumor activities. AB - The synthesis of the unsaturated 4,6-dideoxy-3-fluoro-2-keto-beta-d glucopyranosyl nucleosides of 5-fluorouracil (6a), N(6)-benzoyl adenine (6b), uracil (6c), thymine (6d) and N(4)-benzoyl cytosine (6e), is described. Monoiodination of compounds 1a,b, followed by acetylation, catalytic hydrogenation and finally regioselective 2'-O-deacylation afforded the partially acetylated dideoxynucleoside analogues of 5-fluorouracil (5a) and N(6)-benzoyl adenine (5b), respectively. Direct oxidation of the free hydroxyl group at the 2' position of 5a,b, with simultaneous elimination reaction of the beta-acetoxyl group, afforded the desired unsaturated 4,6-dideoxy-3-fluoro-2-keto-beta-D glucopyranosyl derivatives 6a,b. Compounds 1c-e were used as starting materials for the synthesis of the dideoxy unsaturated carbonyl nucleosides of uracil (6c), thymine (6d) and N(4)-benzoyl cytosine (6e). Similarly a protection-selective deprotection sequence followed by oxidation of the free hydroxyl group at the 2' position of the dideoxy benzoylated analogues 9c-e with simultaneous elimination reaction of the beta-benzoyl group, gave the desired nucleosides 6c-e. None of the compounds was inhibitory to a broad spectrum of DNA and RNA viruses at subtoxic concentrations. The 5-fluorouracil derivative 6a was more cytostatic (50% inhibitory concentration ranging between 0.2 and 12 microM) than the other compounds. PMID- 20018341 TI - Thiophosphorylation of free amino acids and enzyme protein by thiophosphoramidate ions. AB - In search of an activity-preserving protein thiophosphorylation method, with thymidylate synthase recombinant protein used as a substrate, potassium thiophosphoramidate and diammonium thiophosphoramidate salts in Tris- and ammonium carbonate based buffer solutions were employed, proving to serve as a non-destructive environment. Using potassium phosphoramidate or diammonium thiophosphoramidate, a series of phosphorylated and thiophosphorylated amino acid derivatives was prepared, helping, together with computational (using density functional theory, DFT) estimation of (31)P NMR chemical shifts, to assign thiophosphorylated protein NMR resonances and prove the presence of thiophosphorylated lysine, serine and histidine moieties. Methods useful for prediction of (31)P NMR chemical shifts of thiophosphorylated amino acid moieties, and thiophosphates in general, are also presented. The preliminary results obtained from trypsin digestion of enzyme shows peak at m/z 1825.805 which is in perfect agreement with the simulated isotopic pattern distributions for monothiophosphate of TVQQQVHLNQDEYK where thiophosphate moiety is attached to histidine (His(26)) or lysine (Lys(33)) side-chain. PMID- 20018342 TI - Focused toxicity identification evaluations to rapidly identify the cause of toxicity in environmental samples. AB - Over the past 5 years numerous toxicity studies in California using the amphipod, Hyalella azteca, have reported that pyrethroid insecticides, and less frequently the organophosphate insecticide chlorpyrifos, have been responsible for observed toxicity. As work continues to characterize and mitigate these water quality impacts, an approach is needed to screen samples exhibiting toxicity quickly and cost-effectively to establish if these same substances are again responsible, or if other, more atypical toxicants are responsible. Causality is often determined by a standard toxicity identification evaluation (TIE), but when there is strong historical evidence of a likely toxicant, we propose use of a focused TIE procedure to screen samples using manipulations specifically designed to identify pyrethroid- or chlorpyrifos-related toxicity. The focused tests use reduced temperature, piperonyl butoxide addition, and engineered enzymes specifically designed to hydrolyze certain pyrethroid and organophosphate insecticides. The target compounds exhibit unique response profiles to this battery of manipulations, distinct from each other and other non-insecticides. When used in conjunction with analytical chemistry data, the focused approach was successful in identifying cause of toxicity in a variety of urban and agricultural settings. PMID- 20018343 TI - Sequential extraction of polycyclic aromatic hydrocarbons using subcritical water. AB - A rapid sequential subcritical (superheated) water extraction method for polycyclic aromatic hydrocarbons (PAHs) in contaminated soil and sediment is presented. Decreasing the polarity of water by successive increase of the extraction temperature from 50 degrees C to 200 degrees C at the moderate pressure (10.3MPa) enabled selective, non-exhaustive extractions to be performed. Concurrent with increasing temperatures to 150 degrees C there was an increase in PAH extraction efficiencies. For the majority of determinations no significant differences between extractions at 150 degrees C and 200 degrees C were observed. Varied extraction efficiencies of PAHs at the same extraction conditions reflected dissimilarities between environmental matrices investigated. Selective subcritical water extraction of PAHs was proportional to their octanol-water partition coefficients. This technique may be applicable in evaluation of risks associated with PAH contaminated sites and in assessments of their bioremediation potential. PMID- 20018344 TI - Effects of the herbicide glyphosate on biological attributes of Alpaida veniliae (Araneae, Araneidae), in laboratory. AB - In the past decades there has been increasing interest in the study of arthropod predators as effective potential natural enemies to be used in the biological control of agricultural pests. In Argentina, transgenic soybean crops (Round-up Ready, RR) are inhabit by many spider species, some of them in high abundance, being indicative of an import potential for pest predation. This crop is associated with the use of glyphosate, a broad-spectrum herbicide, with low environmental impact, even though since the 80's, several negative effects have been deeply documented on mammals, fishes, amphibians, snails, earthworms, insects, etc. Nowadays, the effects on arthropod physiology, behavior and life history traits as end-points in ecotoxicological evaluations are being recognized. In transgenic soybean crops of Buenos Aires province (Argentina), Alpaida veniliae (Araneae, Araneidae) is one of the most abundant orb web weaver spiders. The purpose of this study was to address the effects of glyphosate on some biological attributes of A. veniliae, in laboratory. Results of this study showed no lethal direct effects of Glifoglex on this spider, but it is the first report in literature about sublethal effects of this herbicide on a spider's biological attributes. Negative effects on prey consumption, web building, fecundity, fertility and developmental time of progeny were observed. Although sublethal effects have received less attention than direct lethal effects, they are relevant from an ecological point of view, since the reduction of the arthropod performance may create risks to arthropod biodiversity conservation in agroecosystems. PMID- 20018345 TI - The evaluation of surface and wastewater genotoxicity using the Allium cepa test. AB - Screening for mutagens in complex environmental mixtures, such as surface water or industrial wastewater, is gradually being accepted as a routine method in environmental monitoring programs. In the present work, the simplified Alliumcepa root assay was utilized to evaluate the possible cyto- and genotoxic effects of surface and wastewaters collected near the Sava River (Croatia) over a three month monitoring period. Physicochemical characterization of the water samples included measurements of conductivity, chemical and biological oxygen demand, levels of suspended matter and salts, nitrate, nitrite, ammonium, total nitrogen and total phosphorus. Morphological modifications of the A. cepa roots, inhibition of root growth, cell division and induction of mitotic and chromosomal aberrations were observed. The most highly polluted water samples (industrial effluents) caused an inhibition of root growth of over 50%, a decrease in the mitotic index of over 40%, and a considerable increase in chromosomal aberrations compared to the control. The measured biological effects of some water samples appeared related to the physicochemical characteristics. Therefore, mutagenicity/genotoxicity assays should be included, along with conventional chemical analysis, in water quality monitoring programs. Their use would allow the quantification of mutagenic hazards in surface and wastewaters. PMID- 20018346 TI - Plasma levels of copper, manganese and selenium in an adult population in southern Spain: influence of age, obesity and lifestyle factors. AB - Copper, manganese and selenium are elements involved in protecting the body against oxidative stress. Determining their plasma level may contribute to assessing the health and nutritional status of populations. The aim of this study was to assess factors influencing copper, manganese and selenium plasma levels in an adult Mediterranean population and to identify groups at risk of deficiency. A cross-sectional survey was carried out in Andalusia, a region in southern Spain. Blood samples were obtained in a random subsample of 340 subjects. Food consumption was assessed by 48-h recall. Height, weight, skinfolds, waist and hip circumferences were measured. Copper, manganese and selenium were measured in plasma. Information about physical exercise, educational level, alcohol and smoking habits was obtained with a structured questionnaire. Plasma copper was found to be higher in women than among men. Hypocupraemia was found in 4.4% of the population, while 9.7% presented hypomanganesemia. Moreover, 86.5% presented plasma selenium values below 125microg/L (cutoff for optimal glutathione peroxidase activity). No association was found between plasma elements, anthropometric indices and lifestyle factors; there were tendencies, no more. Copper tended to decrease in obese and increase in sedentary, while selenium tended to decrease among smokers. Plasma Cu was positively correlated with the consumption of monounsaturated and polyunsaturated fats. Plasma Mn was directly correlated with the consumption of dairy products. Levels of Se were positively correlated with age, the consumption of fruit, vegetables, energy obtained from carbohydrates, and the consumption of fibre, and inversely correlated with the consumption of meat and sweets. Our results provide an estimate of the copper, manganese and selenium status in the adult population of southern Spain. The correlations found for Se suggest that there is a tendency for Se levels to be better maintained among the population that shows a stronger preference for the traditional diet. PMID- 20018347 TI - Monitoring of ochratoxin A exposure of the Portuguese population through a nationwide urine survey--Winter 2007. AB - Ochratoxin A (OTA) is a mycotoxin produced by a variety of fungi, such as Penicillium verrucosum and Aspergillium spp., which has been found to have a wide number of potentially deadly toxic effects, and can enter the human organism through a variety of means. It then finds its way into the bloodstream and, after a lengthy process, is eventually excreted through the urine. It can thus be detected in its original form not only in blood samples but also in this biological medium. As such, and in an attempt to evaluate the exposure of the Portuguese population to this mycotoxin, morning urine samples were collected during the Winter of 2007, from each of five geographically distinct Portuguese locations--Braganca, Porto, Coimbra, Alentejo, and Algarve--and subjected to extraction by immunoaffinity columns and to OTA quantification through liquid chromatography coupled with fluorescence detection. Prevalent incidence was higher than 95% with Coimbra being the exception (incidence of 73.3%). In nearly all locations, the OTA content of most samples was found to be above the limit of quantification (LOQ) of 0.008 ng/ml. Indeed, excluding Coimbra, with an OTA content level of 0.014 ng/ml, all regions featured content values over 0.021 ng/ml. PMID- 20018348 TI - Presence of Integrin alpha(IIb)beta 3 in early gestation human trophoblasts: possible involvement of fibrin as a matrix ligand. AB - INTRODUCTION: Various integrins are expressed on trophoblast membrane and participate in placenta anchoring, cell adhesion , migration and vascular development. Although integrin alpha IIb beta 3 is known to be associated mainly with megakaryocytes and platelets, here we show for the first time its presence in human trophoblasts and in ex-vivo placental villi. METHODS: Trophoblasts were isolated and cultured from early term placentas (9-13 weeks of gestation). The presence of alpha IIb beta 3 integrin was demonstrated in the cells by means of immunofluorescence , flow cytometry and Western blot analysis and in villous placentas by immunohistochemistry. Interaction of trophoblasts with fibronectin and fibrin was evaluated by adhesion assays. RESULTS: Expression of alpha IIb protein in trophoblasts decreased with time in culture and was more prominent when cells were cultured on fibrin as compared to fibronectin. Presence of alpha IIb on villous trophoblasts was documented in placenta sections of 8 and 15 weeks of gestation but not in term placentas. Trophoblasts adhesion to fibrin was reduced by 45% in the presence of blocking antibodies for alpha IIb, but only by 10% to fibronectin, suggesting fibrin as a ligand for trophoblast alpha IIb beta 3. CONCLUSIONS: Our finding demonstrate the transient presence and participation of alpha IIb beta 3 in the orchestrated adhesion molecules of trophblasts and villi. The increased adhesion and expression of alpha IIb in trophoblasts on fibrin suggest its involvement as a ligand in the extracellular milieu of the early fetal placenta. PMID- 20018349 TI - Lymphovascular invasion and pT stage are prognostic factors in patients treated with radical nephroureterectomy for localized upper urinary tract transitional cell carcinoma. AB - OBJECTIVES: To investigate the prognostic significance of lymphovascular invasion (LVI) in patients with localized upper urinary tract transitional cell carcinoma (UUT-TCC) after radical nephroureterectomy. METHODS: The clinical records of 271 patients with UUT-TCC who underwent radical nephroureterectomy between 1986 and 2006 were reviewed. Patients with pT4 stage, lymph node involvement, or distant metastasis were excluded. A total of 238 patients with pTa-3N0M0 were eligible. The prognostic significance of various clinicopathologic factors was analyzed using univariate and multivariate analysis. The mean age was 64.1 years (range, 25-91 years) and the median follow-up duration was 53.4 months (range, 3-240 months). RESULTS: LVI was present in 31 patients (13%). LVI was related to higher pT stage, high tumor grade, sessile architecture, and squamous differentiation. On univariate analysis, tumor architecture, squamous differentiation, LVI, tumor grade, and pT stage influenced disease-specific survival. On multivariate analysis, LVI (hazards ratio [HR], 2.33; P = .014) and pT stage (HR, 2.07; P = .021) showed significantly different rates of disease-specific survival. Patients were classified according to pT stage and LVI. The high-risk group (pT3 and LVI+) showed significantly worse disease-specific survival than the low- (pT < or = 2 and LVI-) or intermediate-risk groups (pT3 and LVI-, pT < or = 2 and LVI+) (P <.001 and P = .032, respectively). CONCLUSIONS: LVI and pT stage are significant prognostic factors for recurrence-free and cancer-specific survivals in patients with localized UUT-TCC. LVI and pT stage would be helpful for selecting patients who are appropriate for postoperative adjuvant chemotherapy. PMID- 20018350 TI - Relationship between body mass index and quantitative 24-hour urine chemistries in patients with nephrolithiasis. AB - OBJECTIVES: To examine the relationship between body mass index and 24-hour urine constituents in a population of stone-forming patients. METHODS: A total of 880 patients who presented to a metabolic stone clinic for initial evaluation were analyzed. Patients were stratified by gender and divided into quartiles of body mass index. Associations between body mass index (BMI) and urine parameters were explored using bivariate and multivariate linear regression. RESULTS: On bivariate analysis, increasing body mass index was associated with a significant increase in sodium, calcium, citrate, uric acid, magnesium, calcium oxalate, uric acid, and a decrease in pH in men. In women, it was associated with a significant increase in sodium, uric acid, oxalate, uric acid, and decreasing pH. On multivariate analysis, BMI was associated only with increases in sodium and calcium oxalate and decrease in pH in men. In women, multivariate analysis demonstrated positive association between BMI and urine sodium, creatinine, and phosphate and a negative relationship with urine citrate and sulfate. CONCLUSIONS: Increasing body mass index was related to several risk factors for urinary stone disease in this study, including increasing urine sodium and decreasing pH in men and increasing urine uric acid, sodium, and decreasing urine citrate in women. Just as general recommendations for patients with nephrolithiasis include high voided volumes, low dietary sodium, and low animal protein intake, perhaps weight reduction should be included as part of the counseling of stone-formers to optimize 24-hour urine parameters. PMID- 20018351 TI - Urology. Letter from the editor. PMID- 20018352 TI - Bladder preservation in octogenarians with invasive bladder cancer. AB - OBJECTIVES: To analyze mortality and morbidity of octogenarians with newly diagnosed invasive transitional cell cancer (TCC) of the bladder who were managed without cystectomy. METHODS: Retrospective chart review of all patients with newly diagnosed invasive TCC (> or = pT1) in the period of 1997-2007, who were 80 years or older at diagnosis. RESULTS: A total of 71 patients (86 + 4 years, mean + standard deviation [SD], pT1: n = 29; > pT2: n = 42) entered this analysis. In this geriatric population, treatment regimens were highly individualized. After transurethral resection, 61% of pT1-patients received bacillus Calmette-Guerin and 62% of those with > pT2-tumors external beam radiation. Mean overall survival (OS) of the entire cohort (n = 71) was 22 + 26 months for pT1-patients 34 + 33 versus 14 + 15 months for those with > or = pT2-tumors (P = .001). Mean cancer specific survival was 58 months for pT1-patients and 11 months for > or = pT2 patients (P <.001). OS was correlated to tumor stage and the degree of mobility, to a lesser extent to the American Society of Anesthesiologists (ASA) score, and only marginally to chronologic age. Satisfactorily bladder function was preserved in 73%. pT1-patients spent 16% of their remaining life-span in the hospital compared with 23% for patients with > pT2-tumors. CONCLUSIONS: OS in TCC is dependent on tumor stage, age, mobility, and comorbidities, and a risk-stratified management is necessary. Patients with pT1G3 tumor and low ASA score have satisfying OS with bladder preservation, but in patients with > or = pT2 and ASA 3-4 the prognosis is very bad. It remains questionable whether patients with tumor stages > or = pT2 and ASA 1-2 despite high age would benefit from radical cystectomy. PMID- 20018354 TI - Vitamin D receptor gene Alw I, Fok I, Apa I, and Taq I polymorphisms in patients with urinary stone. AB - OBJECTIVES: To evaluate vitamin D receptor (VDR) gene polymorphisms in Korean patients so as to identify the candidate genes associated with urinary stones. Urinary stones are a multifactorial disease that includes various genetic factors. METHODS: A normal control group of 535 healthy subjects and 278 patients with urinary stones was evaluated. Of 125 patients who presented stone samples, 102 had calcium stones on chemical analysis. The VDR gene Alw I, Fok I, Apa I, and Taq I polymorphisms were evaluated using the polymerase chain reaction restriction fragment length polymorphism analysis. Allelic and genotypic frequencies were calculated to identify associations in both groups. The haplotype frequencies of the VDR gene polymorphisms for multiple loci were also determined. RESULTS: For the VDR gene Alw I, Fok I, Apa I, and Taq I polymorphisms, there was no statistically significant difference between the patients with urinary stones and the healthy controls. There was also no statistically significant difference between the patients with calcium stones and the healthy controls. A novel haplotype (Ht 4; CTTT) was identified in 13.5% of the patients with urinary stones and in 8.3% of the controls (P = .001). The haplotype frequencies were significantly different between the patients with calcium stones and the controls (P = .004). CONCLUSIONS: The VDR gene Alw I, Fok I, Apa I, and Taq I polymorphisms does not seem to be candidate genetic markers for urinary stones in Korean patients. However, 1 novel haplotype of the VDR gene polymorphisms for multiple loci might be a candidate genetic marker. PMID- 20018353 TI - ERG rearrangement metastasis patterns in locally advanced prostate cancer. AB - OBJECTIVES: To interrogate multifocal prostate cancer (PCa) to determine its predilection for metastasis, using ERG rearrangement as marker of clonality. A hallmark of PCa is that distinct tumor foci may arise independently, which has important biological and clinical implications. Recent studies characterizing ERG rearranged PCa possessing intrafocal homogeneity but interfocal heterogeneity support this hypothesis. METHODS: We studied 26 patients who underwent prostatectomy and lymphadenectomy with at least 2 distinct PCa foci and 1 lymph node (LN) metastasis. Each focus was assessed for size, Gleason score, ERG rearrangement, and TMPRSS2-ERG transcript. RESULTS: Fifteen of 26 cases exhibited interfocal homogeneity with regard to ERG rearrangement (ie, presence vs absence of ERG rearrangement). ERG rearrangement was present in all foci for 6 and absent in all foci for 9 cases. Two cases revealed interfocal heterogeneity with regard to rearrangement mechanism (ie, rearrangement through insertion or deletion). Eight of 26 cases revealed interfocal heterogeneity with regard to rearrangement status. In all cases with at least 1 ERG rearranged focus, we found the corresponding LN metastasis harboring an ERG rearrangement. Interestingly, in a subset of cases the rearrangement status in the LN did not correspond to size or Gleason score. All but 2 ERG rearranged foci had detectable TMPRSS2-ERG transcript levels. CONCLUSIONS: When multifocal PCa demonstrates both ERG positive and ERG-negative foci, the positive foci have a greater predilection for metastasis. Larger studies are needed to confirm the potential additional risk an ERG rearranged focus confers on the likelihood of disease progression. PMID- 20018355 TI - Laparoscopic cryoablation of renal masses: which lesions fail? AB - OBJECTIVE: To identify potential predictive characteristics of lesions which failed in laparoscopic renal cryoablation (LRC). We analyzed 47 lesions that underwent this approach. METHODS: We reviewed 45 consecutive patients who underwent LRC of a renal mass between 2003 and 2008 at a single institution. A total of 47 masses were identified; all were treated by 2 surgeons. We analyzed patient age, ASA, pre- and postoperative creatinine, tumor size, location, number of cryoprobes used, and histology of the lesions. We reviewed imaging to identify characteristics of those lesions which failed LRC management, defined as lesions that demonstrated persistent enhancement and/or did not decrease in size within 6 months of therapy. RESULTS: A total of 47 lesions in 45 patients were identified. The median follow-up was 13 months. Mean lesion size was 2.7 cm (range, 1.2-5.4), with 25 anterior and 22 lateral or posterior lesions. Of the biopsy samples from 40 of 47 lesions, renal cell carcinoma was found in 23, oncocytoma was found in 7, and 10 were benign or inconclusive. Treatment failure was noted in 8 of 47 lesions (17%), 7 of which (87.5% of failed lesions) had broad-based contact with the renal sinus. Broad-based lesions which made contact with the renal sinus were successfully treated 53.3% of the time, whereas lesions which lacked contact with the renal sinus were treated successfully 96.9% of the time (P <.01). CONCLUSIONS: Broad-based central location of a renal mass may predict a significantly increased risk of failure of LRC and should be considered in patient counseling. PMID- 20018356 TI - Polyostotic fibrous dysplasia mimicking bone metastases in a patient with advanced-stage cervical cancer. PMID- 20018357 TI - Characterization of ram (Ovis aries) sperm head morphometry using the Sperm-Class Analyzer. AB - Sperm morphology has been identified as a characteristic that can be useful in the prediction of fertilizing capacity. The aim of the current study was to characterize ram sperm heads morphometrically as a basis for future studies on the relationship between sperm quality and male fertility. For this purpose, ejaculates from 241 mature rams (Ovis aries) belonging to 36 different dairy herds were used to evaluate sperm head morphometry by means of the Sperm-Class Analyzer. Sperm samples, collected by artificial vagina, were diluted in phosphate buffered saline (PBS) for the analysis. A microscope slide was prepared from single-diluted fresh sperm samples. Slides were air-dried and stained with Hemacolor. A minimum of 115 sperm heads were analyzed from each male. Each sperm head was measured for four primary parameters (area, perimeter, length, width), and four derived parameters of head shape were obtained. Significant differences in sperm head morphometry were found between rams (CV for morphometric parameters ranging from 0.9 to 10.1), and there were marked differences in the sperm morphometric composition of the ejaculates. For all parameters, within-animal CVs were greater than between-animal CVs. Within-animal CVs ranged from 4.2 to 10.6, showing the high degree of sperm polymorphism present in the sheep ejaculate. Significant differences in sperm head morphometry were found between rams belonging to the different herds (i.e., origin). An important part of the variability observed on morphometric parameters was due to the male itself, with an explained variance ranging from 3.6% for regularity to 34.0% for p2a (perimeter(2)/[4xpixarea]). The explained variance by the herd of origin of the males ranged from 0.6% for regularity to 10.8% for area. Our results suggest that a genetic component might be responsible for the observed sperm head morphometry differences between herds. PMID- 20018358 TI - Ovarian stimulation with follicle-stimulating hormone under increasing or minimal concentration of progesterone in dairy cows. AB - The objective of this study was to investigate the effect of the presence or absence of Corpus luteum (CL) on the follicular population during superstimulation in dairy cows (Holstein-Friesian cattle). Animals were divided into two groups as follows: (1) Growing CL group (G1): Cows (n=7) received a total dose of 28 Armour units (AU) follicle-stimulating hormone (FSH) through the first 4 d (twice daily) after spontaneous ovulation (Day 0). (2) CL Absence group (G2): Cows (n=10) received prostaglandin F(2alpha) (PGF(2alpha)) at 9 or 10 d after ovulation. After 36h, all the follicles (larger than 5mm) were aspirated (Day 0). The FSH treatment started 24h after aspiration and continued for 4 d. The number of small (3 to <5mm), medium (5 to <8mm), and large (> or = 8mm) follicles was examined on Days 1, 3, and 5 in all groups. Blood samples were collected daily for 5 d, and progesterone (P(4)), estradiol (E(2)), insulin-like growth factor-1 (IGF-1), and growth hormone (GH) in plasma were measured by enzyme immunoassays. The results showed that in G1, the P(4) level increased gradually from 0.5 ng/mL at Day 1 to 2 ng/mL at Day 5, whereas in G2, the P(4) level was completely below 0.5 ng/mL. All cows of the G2 group showed an increase of E(2) at Day 3 or Day 4 followed by an increase of IGF-1 within 24h, while GH increased concomitantly with the E(2) increase in 8 of 10 trials. On the other hand, cows of the G1 group showed neither E(2) nor IGF-1 increase. Moreover, at the end of the treatment, the number of follicles in the G2 group was significantly increased compared with that of the G1 group (22.8+/-2.0 vs. 11.6+/ 2.0). In conclusion, low P(4) level during FSH treatment enhanced multiple follicular growth and E(2) secretion, which was followed by increase of IGF-1 and GH. Therefore, the absence of the CL may play a critical role in the superovulation response by controlling the number of growing follicles. PMID- 20018359 TI - Freezing dog semen in presence of the antioxidant butylated hydroxytoluene improves postthaw sperm membrane integrity. AB - In an attempt to evaluate the protective effect of a lipid-soluble antioxidant (butylated hydroxytoluene; BHT), semen from four dogs (Canis familiaris) was frozen in two different extenders (Uppsala or INRA-96 plus glycerol) with or without 1mM BHT. Sperm membrane integrity using flow cytometry and motility using a computerized system were evaluated in each experimental group. The Uppsala extender was superior in all aspects of sperm function. The percentage of sperm membranes was significantly higher in semen samples frozen in presence of BHT. Our results suggest that the Uppsala extender can be improved with the addition of BHT. PMID- 20018360 TI - Common and segregated neural substrates for automatic conceptual and affective priming as revealed by event-related functional magnetic resonance imaging. AB - The brain activity associated with automatic semantic priming has been extensively studied. Thus far there has been no prior study that directly contrasts the neural mechanisms of semantic and affective priming. The present study employed event-related fMRI to examine the common and distinct neural bases underlying conceptual and affective priming with a lexical decision task. A special type of emotional word, a dual-meaning word containing both conceptual meaning and affective meaning, was adopted as target. Short stimulus onset asynchrony (SOA) (50 ms) was used to emphasize automatic processing. Fifteen participants were scanned in the present study. We found that the left middle/superior temporal gyrus was the brain region involved in both automatic conceptual and affective priming effects, suggesting general lexical-semantic processing that share in the two types of priming. The left inferior frontal gyrus and right superior temporal gyrus were found to be the conceptual-specific areas in automatic priming effect, consistent with the role of these areas in more extensive within-category semantic processes. The results also revealed that the left fusiform gyrus and left insula were the affective-specific regions in automatic priming effect, demonstrating the involvement of the left fusiform gyrus in automatic affective priming effect, and clarifying the role of the insula in emotional processing rather than conceptual processing. Despite comparable behavioral effects of automatic conceptual priming and affective priming, the present study revealed a neural dissociation of the two types of priming, as well as the shared neural bases. PMID- 20018361 TI - Surface characterization of zirconia dental implants. AB - OBJECTIVES: The aim of the study was to characterize the chemical composition, microstructure and roughness of two commercially available zirconia dental implants (WhiteSky and Zit-Z). METHODS: The chemical composition of the cervical collar and threaded root parts of the implants (n=2) were studied by XPS and HV EDX. LV-SEM was used for morphological assessment, Raman microanalysis for microstructural characterization and optical profilometry for surface roughness measurements. XRD, HV-EDX and Raman microanalysis of bulk regions (longitudinal sections) were used as reference. RESULTS: XPS showed the presence of C, O, Zr and Y (collar) plus Al (root) at implant surfaces. More C (10-26at%) and a lower Al/Zr ratio were found in WhiteSky (1.05 vs 1.26 in Zit-Z). Zr, Y and Al were detected in single, fully oxidized states. The same elements, plus Hf, were identified by HV-EDX at bulk and surface regions, with a Al/Zr ratio higher in WhiteSky (0.17 vs 0.09 in Zit-Z). Na, K and Cl contaminants were traced at implant root parts by both methods. XRD analysis of cross-sectioned specimens revealed the presence of monoclinic and tetragonal zirconia along with cubic yttria phases. Raman microanalysis showed that the monoclinic zirconia volume fraction was higher at root surfaces than the collar. No monoclinic phase was found at bulk regions. Significantly higher Sa and Sq values were recorded in WhiteSky than Zit-Z, whereas Zit-Z showed higher Rt value. SIGNIFICANCE: The differences found between the implants in the extent of carbon contamination, residual alumina content, tetragonal to monoclinic ZrO(2) phase transformation and 3D-roughness parameters may contribute to a substantial differentiation in the cellular and tissue response. PMID- 20018362 TI - A review of adhesion science. AB - OBJECTIVE: Adhesion or cohesion includes an adherend, adhesive, and intervening interface. Adhesive joints may include one or more interfaces. Adhesion science focuses on understanding the materials properties associated with formation of the interfaces, changes in the interfaces with time, and events associated with failure of the interfaces. METHODS: The key principles for good interface formation are creation of a clean surface, generation of a rough surface for interfacial interlocking, good wetting of the substratum by the adhesive/cohesive materials, adequate flow and adaptation for intimate interaction, and acceptable curing when phase changes are required for final joint formation. RESULTS: Much more effort is needed in the future to carefully assess each of these using available testing methods that attempt to characterize the energetics of the interfaces. Bonding involves potential contributions from physical, chemical, and mechanical sources but primarily relies on micro-mechanical interaction for success. Characterization of the interface before adhesion, during service, and after failure would be much more useful for future investigations and remains as a great challenge. SIGNIFICANCE: Scientists should more rigorously apply techniques such as comprehensive contact angle analysis (rather than simple water wettability) for surface energy determination, and AFM in addition to SEM for surface texture analysis. PMID- 20018363 TI - Influence of different initiators on the degree of conversion of experimental adhesive blends in relation to their hydrophilicity and solvent content. AB - OBJECTIVES: The aim of this study was to compare the curing reaction of five experimental adhesive blends containing different photo-initiating systems. The hypothesis tested was that degree of conversion (DC) of resin blends is affected by resin type, solvent content and photo-initiating system. METHODS: The experimental methacrylate resin blends were ranked from hydrophobic (R2) to hydrophilic (R3 and R4) and tested as neat, or solvated with 10% or 20% ethanol, or 10% ethanol and 10% water. Three different photo-initiators were used: IS 1=0.25% CQ (camphorquinone)+1% EDMAB (ethyl 4-dimethylaminobenzoate); IS-2=1.25% TPO (diphenyl(2,4,6-trimethylbenzoyl)-phosphine oxide); IS-3=0.25% CQ+0.50% EDMAB+0.50% TPO. DC of resin blends was measured with a differential scanning calorimeter. Data were analyzed with a three-way ANOVA. RESULTS: Neat resin type influenced DC, as R4 showed the highest values compared to R2 and R3 (p<0.05). Solvent had a significant effect on DC (p<0.05): dilution of resin blends with 10% or 20% ethanol or 10% ethanol+10% water increased the DC of all resins, except for R4. Initiators influenced the polymerization since neat resins and mixtures solvated with 10% or 20% ethanol showed their highest DC values when polymerized with IS-1 or IS-3 (p>0.05), while IS-2 or IS-3 increased the DC values of resins diluted with 10% ethanol and 10% water (p<0.05). CONCLUSIONS: Water-compatible photo-initiators such as TPO should be included in the hydrophilic solvated adhesive formulation to ensure an appropriate DC of the adhesive layer. PMID- 20018364 TI - Safety and immunogenicity of 2009 pandemic influenza A H1N1 vaccines in China: a multicentre, double-blind, randomised, placebo-controlled trial. AB - BACKGROUND: The current influenza pandemic calls for a safe and effective vaccine. We assessed the safety and immunogenicity of eight formulations of 2009 pandemic influenza A H1N1 vaccine produced by ten Chinese manufacturers. METHODS: In this multicentre, double-blind, randomised trial, 12 691 people aged 3 years or older were recruited in ten centres in China. In each centre, participants were stratified by age and randomly assigned by a random number table to receive one of several vaccine formulations or placebo. The study assessed eight formulations: split-virion formulation containing 7.5 microg, 15 microg, or 30 microg haemagglutinin per dose, with or without aluminium hydroxide adjuvant, and whole-virion formulation containing 5 microg or 10 microg haemagglutinin per dose, with adjuvant. All formulations were produced from the reassortant strain X 179A (A/California/07/2009-A/PR/8/34). We analysed the safety (adverse events), immunogenicity (geometric mean titre [GMT] of haemagglutination inhibition antibody), and seroprotection (GMT >or=1:40) of the formulations. Analysis was by per protocol. Two sites registered their trial with ClinicalTrials.gov, numbers NCT00956111 and NCT00975572. The other eight studies were registered with the State Food and Drug Administration of China. FINDINGS: 12 691 participants received the first dose on day 0, and 12 348 participants received the second dose on day 21. The seroprotection rate 21 days after the first dose of vaccine ranged from 69.5% (95% CI 65.9-72.8) for the 7.5 microg adjuvant split-virion formulation to 92.8% (91.9-93.6) for the 30 microg non-adjuvant split-virion formulation. The seroprotection rate was 86.5% (796 of 920; 84.1-88.7) in recipients of one dose of the 7.5 microg non-adjuvant split-virion vaccine compared with 9.8% (140 of 1432; 8.3-11.4) in recipients of placebo (p<0.0001). One dose of the 7.5 microg non-adjuvant split-virion vaccine induced seroprotection in 178 of 232 children (aged 3 years to <12 years; 76.7%, 70.7 82.0), 211 of 218 adolescents (12 years to <18 years; 96.8%, 93.5-98.7), 289 of 323 adults (18-60 years; 89.5%, 85.6-92.6), and 118 of 147 adults older than 60 years (80.3%, 72.9-86.4), meeting the European Union's licensure criteria for seroprotection in all age-groups. In children, a second dose of the 7.5 microg formulation increased the seroprotection rate to 97.7% (215 of 220, 94.8-99.3). Adverse reactions were mostly mild or moderate, and self-limited. Severe adverse effects occurred in 69 (0.6%, 0.5-0.8) recipients of vaccine compared with one recipient (0.1%, 0-0.2) of placebo. The most common severe adverse reaction was fever, which occurred in 25 (0.22%; 0.14-0.33) recipients of vaccine after the first dose and four (0.04%; 0.01-0.09) recipients of vaccine after the second dose compared with no recipients of placebo after either dose. INTERPRETATION: One dose of non-adjuvant split-virion vaccine containing 7.5 microg haemagglutinin could be promoted as the formulation of choice against 2009 pandemic influenza A H1N1 for people aged 12 years or older. In children (aged <12 years), two 7.5 mug doses might be needed. FUNDING: Sinovac Biotech, Hualan Biological Bacterin, China National Biotec Group, Beijing Tiantan Biological Products, Changchun Institute of Biological Products, Changchun Changsheng Life Sciences, Jiangsu Yanshen Biological Technology Stock, Zhejiang Tianyuan Bio Pharmaceutical, Lanzhou Institute of Biological Products, Shanghai Institute of Biological Products, and Dalian Aleph Biomedical. PMID- 20018365 TI - Immune response after a single vaccination against 2009 influenza A H1N1 in USA: a preliminary report of two randomised controlled phase 2 trials. AB - BACKGROUND: Data are needed from large clinical trials of paediatric, adult, and elderly people to find the appropriate antigen dose and vaccination schedule for the 2009 pandemic influenza A H1N1. We therefore report preliminary safety and immunogenicity results after one injection of a licensed monovalent pandemic H1N1 vaccine in the USA. METHODS: We randomly assigned healthy children (aged 6-35 months and 3-9 years) and adults (18-64 years and >or=65 years) to vaccine containing per dose 7.5 microg (children and adults), 15 microg (children and adults), or 30 microg (adults only) haemagglutinin in two placebo-controlled, observer-masked, multicentre phase 2 studies done in the USA. Participants were allocated with an interactive voice-response system or computer-generated randomisation lists with opaque scratchable patches. Primary outcome was haemagglutination inhibition antibody response 21 days after the first of two planned vaccinations (interim analysis of studies in progress). Analyses were by full-analysis set. The trials are registered with ClinicalTrials.gov as NCT00953524 and NCT00952419. FINDINGS: 410 of 423 children and 724 of 750 adults given an active vaccine, and 50 of 51 children and 95 of 99 adults given placebo were assessed for immunogenicity on day 21. After active vaccination, 45 of 101 (45%; 95% CI 35-55) to 47 of 94 (50%; 40-61) infants aged 6-35 months, 75 of 109 (69%; 59-77) to 80 of 106 (75%; 66-83) 3-9-year-old children, 134 of 141 (95%; 90 98) to 144 of 144 (100%; 98-100) of 18-64-year-old adults, and 93 of 100 (93%; 86 96) to 93 of 98 (95%; 89-98) elderly adults were seroprotected (proportion with titres >or=1:40). No vaccine-related serious adverse events occurred. Injection site and systemic reactions were reported by up to about 50% of every age and vaccine group, with no noticeable differences between vaccine and placebo groups. INTERPRETATION: One dose of vaccine was highly immunogenic in adults, suggesting that it afforded sufficient protection against this pandemic influenza A H1N1 virus. Two doses of vaccine will probably be needed in children younger than 9 years. Safety and reactogenicity of the vaccine were acceptable and similar to those of seasonal vaccine. FUNDING: Office of the Assistant Secretary for Preparedness and Response, and Biomedical Advanced Research and Development Authority. PMID- 20018366 TI - Large trials confirm immunogenicity of H1N1 vaccines. PMID- 20018367 TI - Safety and immunogenicity of a 2009 pandemic influenza A H1N1 vaccine when administered alone or simultaneously with the seasonal influenza vaccine for the 2009-10 influenza season: a multicentre, randomised controlled trial. AB - BACKGROUND: With the ongoing 2009 pandemic of influenza A H1N1, development of pandemic influenza vaccines has generated much interest. We investigated the safety and immunogenicity of a whole-virion, inactivated, adjuvanted pandemic H1N1 vaccine in adult and elderly volunteers, given without or simultaneously with the 2009-10 seasonal trivalent influenza vaccine. METHODS: This prospective, randomised study was undertaken in two centres in Hungary. 355 participants, including 203 adults (18-60 years) and 152 elderly people (>60 years), were assigned by stratified randomisation to either 0.5 mL of the pandemic vaccine (Fluval P, a monovalent vaccine with 6 microg haemagglutinin per 0.5 mL content and aluminium phosphate gel adjuvant; n=178) or 0.5 mL of the pandemic vaccine and 0.5 mL of the seasonal trivalent vaccine (Fluval AB, a trivalent inactivated whole-virion influenza vaccine; n=177). All vaccinations were done by specific study personnel, who did not take part in the assessment of safety or immunogenicity. Co-primary objectives were safety and immunogenicity by haemagglutinin inhibition testing. All analyses were done according to a pre established analysis plan. This study is registered with ClinicalTrials.gov, number NCT01010893. FINDINGS: Two participants receiving the pandemic vaccine only (group 1) and one receiving pandemic and seasonal vaccines (group 2) were lost to follow-up. Participants in both groups developed antibody responses against the pandemic influenza A H1N1 virus (group 1: seroconversion for adults 74.3%, 95% CI 64-6-82.4 and for elderly people 61.3%, 49.1-72.4; group 2: 76.8%, 67.2-84.7 and 81.8%, 71.4-89.7, respectively). Single doses of 6 microg fulfilled European Union and US licensing criteria for interpandemic and pandemic influenza vaccines. Simultaneously, participants in group 2 developed the immune responses needed for licensing for all three seasonal strains in the seasonal vaccine for the 2009-10 season. All adverse events were rare, mild, and transient; the most frequent were pain at injection site (eight cases in group 1 vs 18 in group 2) and fatigue for 1-2 days after vaccination (three vs five cases). INTERPRETATION: The present pandemic vaccine is safe and immunogenic in healthy adult and elderly patients, and needs low doses and only one injection to trigger immune responses to comply with licensing criteria. It can be safely co-administered with the 2009 10 seasonal influenza vaccine. FUNDING: Omninvest, Hungary. PMID- 20018368 TI - Defining the safety profile of pandemic influenza vaccines. PMID- 20018369 TI - Self-assembling methoxypoly(ethylene glycol)-b-poly(carbonate-co-L-lactide) block copolymers for drug delivery. AB - Bicalutamide is the most widely used non-steroidal antiandrogen for treating early stage prostate cancer, but suffers variable oral absorption due to its limited aqueous solubility. Thus, our objective was to synthesize novel biodegradable copolymers for the systemic micellar delivery of bicalutamide. Flory-Huggins interaction parameter (cFH) was used to assess compatibility between bicalutamide and poly(L-lactide) or poly(carbonate-co-lactide) polymer pairs. Polyethylene glycol-b-poly(carbonate-co-lactide) [PEG-b-P(CB-co-LA)] copolymers were synthesized and characterized by NMR and gel permeation chromatography. These micelles had average diameter of 100 nm and had a smooth surface and distinct spherical shape. Drug loading studies revealed that adding the carbonate monomer could increase bicalutamide loading. Among the series, drug loading of micelles formulated with PEG-b-P(CB-co-LA) copolymer containing 20 mol% carbonate was about four-fold higher than PEG-b-PLLA and aqueous solubility of bicalutamide increased from 5 to 4000 microg/mL. CMC values for PEG-b-P(CB-co LA) copolymers was up to 10-fold lower than those of PEG-b-PLLA. In vitro release experiments showed PEG-b-P(CB-co-LA) copolymers to be more efficient in sustaining the release of bicalutamide compared to PEG-b-PLLA. Bicalutamide loaded PEG-b-P(CB-co-LA) micelles showed significant inhibition of LNCaP cell growth in a dose-dependent manner which was similar to the methanol solution of free drug. PMID- 20018370 TI - Expression and distribution of tight junction proteins in human amnion during late pregnancy. AB - Amnion is the innermost layer of the fetal membrane and has been suggested to regulate the volume of amniotic fluid via the amniotic epithelium. The transepithelial pathway is generally restricted by tight junctions (TJs). Thus far, human amniotic TJs have not been identified. In this study, we determined whether the human amniotic epithelium contains TJs. Reverse transcription polymerase chain reaction (RT-PCR) and western blotting analyses showed that the human amniotic epithelium has TJ components, such as occludin, ZO-1, and at least 2 types of claudins, i.e., claudin-4 and claudin-7. The TJ components were found to localize in the lateral membranes and cytoplasm at 35 weeks of gestation; these components disappeared from the lateral membrane at 37 weeks of gestation. Organ culturing of the amnion at 37 weeks gestation induced the relocalization of the TJ proteins from the cytoplasm to the lateral membranes. Furthermore, in cultured amniotic epithelial cells, dexamethasone induced the downregulation of the protein expression of TJs. These findings suggest that the human amniotic epithelium has TJs that disrupt during late pregnancy. The disruption may be induced by several factors such as glucocorticoids present in the amniotic fluid during late pregnancy. PMID- 20018371 TI - Arachidonic acid stimulates extracellular Ca(2+) entry in rat pancreatic beta cells via activation of the noncapacitative arachidonate-regulated Ca(2+) (ARC) channels. AB - Arachidonic acid (AA) is generated in the pancreatic islets during glucose stimulation. We investigated whether AA activated extracellular Ca(2+) entry in rat pancreatic beta cells via a pathway that was independent of the activation of voltage-gated Ca(2+) channels. The AA triggered [Ca(2+)](i) rise did not involve activation of GPR40 receptors or AA metabolism. When cells were voltage clamped at -70mV, the AA-mediated intracellular Ca(2+) release was accompanied by extracellular Ca(2+) entry. AA accelerated the rate of Mn(2+) quench of indo-1 fluorescence (near the Ca(2+)-independent wavelength of indo-1), reflecting the activation of a Ca(2+)-permeable pathway. The AA-mediated acceleration of Mn(2+) quench was inhibited by La(3+) but not by 2-APB (a blocker of capacitative Ca(2+) entry), suggesting the involvement of arachidonate-regulated Ca(2+) (ARC) channels. Consistent with this, intracellular application of the charged membrane impermeant analog of AA, arachidonyl-coenzyme A (ACoA) triggered extracellular Ca(2+) entry, as well as the activation of a La(3+)-sensitive small inward current (1.7pA/pF) at -70mV. Our results indicate that the activation of ARC channels by intracellular AA triggers extracellular Ca(2+) entry. This action may contribute to the effects of AA on Ca(2+) signals and insulin secretion in rat beta cells. PMID- 20018372 TI - Succinate modulates Ca(2+) transient and cardiomyocyte viability through PKA dependent pathway. AB - GPR91 is an orphan G-protein-coupled receptor (GPCR) that has been characterized as a receptor for succinate, a citric acid cycle intermediate, in several tissues. In the heart, the role of succinate is unknown. We now report that rat ventricular cardiomyocytes express GPR91. We found that succinate, through GPR91, increases the amplitude and the rate of decline of global Ca(2+) transient, by increasing the phosphorylation levels of ryanodine receptor and phospholamban, two well known Ca(2+) handling proteins. The effects of succinate on Ca(2+) transient were abolished by pre-treatment with adenylyl cyclase and cAMP dependent protein kinase (PKA) inhibitors. Direct PKA activation by succinate was further confirmed using a FRET-based A-kinase activity reporter. Additionally, succinate decreases cardiomyocyte viability through a caspase-3 activation pathway, effect also prevented by PKA inhibition. Taken together, these observations show that succinate acts as a signaling molecule in cardiomyocytes, modulating global Ca(2+) transient and cell viability through a PKA-dependent pathway. PMID- 20018373 TI - Complexity of miR-223 regulation by CEBPA in human AML. AB - microRNA-223 (miR-223) can trigger normal granulopoiesis. miR-223 expression is regulated by two distinct CEBPA (CCAAT/enhancer binding protein-alpha) sites. Here, we report that miR-223 is largely suppressed in cells from acute myeloid leukemia (AML) patients. By sequencing, we found that miR-223 suppression in AML is not caused by DNA sequence alterations, nor is it mediated by promoter hypermethylation. The analysis of the individual contribution of both CEBPA sites to miR-223 regulation identified the site upstream of the miR-223 primary transcript as the predominant regulatory element. Our results suggest that miR 223 suppression in AML is caused by impaired miR-223 upstream factors. PMID- 20018374 TI - Modifications of fasting glucose values as first sign of resistance in chronic myeloid leukemia chronic phase patients during imatinib treatment. PMID- 20018375 TI - A myeloma translocation-like model associating CCND1 with the immunoglobulin heavy-chain locus 3' enhancers does not promote by itself B-cell malignancies. AB - Cyclin D1 overexpression is associated with mantle cell lymphoma and multiple myeloma. In myeloma, it often results from chromosomal translocations linking the CCND1 gene to the 3' part of the IgH locus constant region. This region includes a single and potent transcriptional regulatory region (RR) 3' of the Calpha gene mostly active in mature B-cells. To check whether this RR alone was sufficient to deregulate CCND1, we generated mice carrying a 3'IgH RR-driven human CCND1 transgene and specifically up-regulating cyclin D1 expression in B-cells. In transgenic B-cells, cyclin D1 enforced cell cycle entry in response to various stimuli (LPS, anti-IgM, anti-CD40) but also increased cell death, so that exaggerated proliferation did not result in peripheral lymphocytosis. Despite exaggerated B-cell entry into G(1) phase, malignant lymphoproliferation did not occur either. Crossing of CCND1-3'IgH RR mice with c-myc-3'IgH RR mice did not reveal accelerated tumorigenesis as compared with c-myc-3'IgH RR mice alone. The data presented here demonstrate that the 3'IgH RR-mediated deregulation of CCND1 in mature B-cells cannot by itself trigger the development of lymphomas and strengthen the concept that cyclin D1 per se is not an armful proto-oncogene. Rather its overexpression in several malignancies might be only a stigma of lymphomagenesis or represent a single hit within a multiple hit process. PMID- 20018376 TI - Engineered regulatory T cells prevent graft-versus-host disease while sparing the graft-versus-leukemia effect after bone marrow transplantation. AB - Regulatory T cells (Tregs) can prevent graft-versus-host disease (GVHD) after allogeneic bone marrow transplantation (BMT). Here we developed a lentivirus based strategy to ectopically express Foxp3 in mouse CD4(+)CD25(-) T cells. These cells shared similar immunophenotypes and biological features of natural Tregs. Co-injection of engineered Tregs with donor bone marrow cells and splenocytes prevented recipients from lethal GVHD. Furthermore, we showed that graft-versus leukemia (GVL) effect against EL4/DsRed leukemic cells was maximally preserved while GVHD was minimized during exposure to engineered Tregs in a mouse leukemia model. These findings provide a novel approach to preventing GVHD while maintaining GVL effect during BMT. PMID- 20018377 TI - Molecular identification of airborne bacteria associated with aerial spraying of bovine slurry waste employing 16S rRNA gene PCR and gene sequencing techniques. AB - Polymerase chain reaction amplification of the universal 16S ribosomal RNA (rRNA) gene was performed on a collection of 38 bacterial isolates, originating from air sampled immediately adjacent to the agricultural spreading of bovine slurry. A total of 16 bacterial genera were identified including both Gram-positive and Gram-negative genera. Gram-positive organisms accounted for 34/38 (89.5%) of total bacterial numbers consisting of 12 genera and included Staphylococcus (most common genus isolated), Arthrobacter (2nd most common genus isolated), Brachybacterium, Exiguobacterium, Lactococcus, Microbacterium and Sporosarcina (next most common genera isolated) and finally, Bacillus, Brevibacterium, Frigoribacterium, Mycoplana and Pseudoclavibacter. Gram-negative organisms accounted for only 4/38 (10.5%) bacterial isolates and included the following genera, Brevundimonas, Lysobacter, Psychrobacter and Rhizobium. No gastrointestinal pathogens were detected. Although this study demonstrated a high diversity of the microorganisms present, only a few have been shown to be opportunistically pathogenic to humans and none of these organisms described have been described previously as having an inhalational route of infection and therefore we do not believe that the species of organisms identified pose a significant health and safety threat for immunocompetant individuals. PMID- 20018378 TI - Mycobacterial antigen(s) induce anergy by altering TCR- and TCR/CD28-induced signalling events: insights into T-cell unresponsiveness in leprosy. AB - Present study investigates the role of Mycobacterium leprae (M. leprae) antigens on TCR- and TCR/CD28-induced signalling leading to T-cell activation and further correlates these early biochemical events with T-cell anergy, as prevailed in advanced stages of leprosy. We observed that both whole cell lystae (WCL) and soluble fraction of M. leprae sonicate (MLSA) not only inhibited TCR, thapsigargin and ionomycin induced calcium fluxes by diminishing the opening of calcium channels, but also TCR- or TCR/CD28-induced proximal signalling events like phosphorylation of Zap-70 and protein kinase-C (PKC) activity. Study of TCR- and TCR/CD28-induced downstream signals revealed that M. leprae antigens curtail phosphorylation of both Erk1/2 and p38MAPK, consequently altering terminal signalling events like reduced binding of NFAT on IL-2 promoter and transcription of IL-2 gene, diminished expression of activation markers (CD25 and CD69). Furthermore, M. leprae fractions significantly inhibited IL-2 secretion and T cell blastogenesis in healthy individuals. Altogether, results suggest that M. leprae interferes with TCR/CD28-induced upstream as well as downstream signalling events resulting in reduced IL-2 production and thus inhibition in T-cell proliferation, which might be responsible for T-cell unresponsiveness leading to stage of immunosuppression and consequently, for the progression of disease. PMID- 20018379 TI - Limbal stem cell deficiency in chronic and delayed-onset mustard gas keratopathy. AB - PURPOSE: To evaluate limbal stem cell deficiency (LSCD) using impression cytology in patients with chronic and delayed-onset mustard gas keratopathy (MGK). DESIGN: Prospective observational case series. PARTICIPANTS: Thirty-five eyes of 18 patients (all male) with MGK were included. METHODS: A consecutive series of patients with MGK underwent impression cytology. Finding of goblet cells on the corneal side of specimens was considered as LSCD. Severity of corneal clinical manifestation was graded as mild, moderate, and severe in each quadrant. Relation between impression cytology findings and clinical grading was evaluated. MAIN OUTCOME MEASURES: Impression cytology findings and clinical grading. RESULTS: There was LSCD in at least 1 quadrant of cornea in all 35 eyes (100% of cases). No differences were found between impression cytology findings (positive vs. negative for corneal goblet cells) among different quadrants (P = 0.378). Clinical grading was the same between nasal and temporal quadrants (P = 0.266) and between superior and inferior quadrants (P = 0.263). By combining superior and inferior quadrants (vertical zone) and nasal and temporal quadrants (horizontal zone), corneal clinical grading was more severe in horizontal versus vertical zones (P<0.001). There was no relation between LSCD and corneal clinical severity (P = 0.893). CONCLUSIONS: A varying degree of LSCD was demonstrated in all patients with chronic or delayed-onset MGK using impression cytology. Corneal clinical manifestations are more severe in nasal and temporal quadrants. There was no relation between impression cytology findings (positive vs. negative for goblet cells) and corneal clinical grading. Other factors, such as perilimbal conjunctival ischemia, may play a role. PMID- 20018380 TI - Self-reported use of eye care among Latinos: the Los Angeles Latino Eye Study. AB - PURPOSE: To identify the prevalence and determinants of self-reported eye care use in Latinos. DESIGN: Population-based ocular epidemiologic study in Latinos aged 40+ years living in La Puente, California. PARTICIPANTS: A total of 5455 participants. METHODS: Univariate, multivariable, and stepwise logistic regression analyses were conducted to identify predisposing, enabling, and need variables associated with self-reported eye care use. MAIN OUTCOME MEASURES: Prevalence of self-reported use: eye care visit, having had a dilated examination in the past 12 months, ever having had a dilated examination, and odds ratios for factors associated with self-reported use. RESULTS: Overall, 36% of participants reported an eye care visit and 19% reported having a dilated examination in the past year. Fifty-seven percent reported ever having had a dilated eye examination. Greater eye care use was associated with older age, female gender, bilingual language proficiency (English and Spanish), more education, having health insurance, having a usual place for care, having a regular provider of care, a greater number of comorbidities, visual impairment, and lower vision specific quality of life scores. CONCLUSIONS: Multiple modifiable factors are associated with greater use and access to eye care for Latinos. Modification of these factors should be a priority because visual impairment has significant impacts on well-being and mortality. PMID- 20018381 TI - Volumetric MRI changes, cognition and personality traits in old age depression. AB - BACKGROUND: The presence of cognitive and structural deficits in euthymic elderly depressed patients remains a matter of debate. Integrative aetiological models assessing concomitantly these parameters as well as markers of psychological vulnerability such as persistent personality traits, are still lacking for this age group. METHODS: Cross-sectional comparisons of 38 elderly remitted patients with early-onset depression (EOD) and 62 healthy controls included detailed neuropsychological assessment, estimates of brain volumes in limbic areas and white matter hyperintensities, as well as evaluation of the Five-Factor personality dimensions. RESULTS: Both cognitive performances and brain volumes were preserved in euthymic EOD patients. No significant group differences were observed in white matter hyperintensity scores between the two groups. In contrast, EOD was associated with significant increase of Neuroticism and decrease of Extraversion facet scores. LIMITATIONS: Results concern the restricted portion of EOD patients without psychiatric and physical comorbidities. Future longitudinal studies are necessary to determine the temporal relationship between the occurrence of depression and personality dimensions. CONCLUSIONS: After remission from acute depressive symptoms, cognitive performances remain intact in elderly patients with EOD. In contrast to previous observations, these patients display neither significant brain volume loss in limbic areas nor increased vascular burden compared to healthy controls. Further clinical investigations on EOD patterns of vulnerability in old age will gain from focusing on psychological features such as personality traits rather than neurocognitive clues. PMID- 20018382 TI - A specific induction of interleukin-10 by the Map41 recombinant PPE antigen of Mycobacterium avium subsp. paratuberculosis. AB - Interleukin-10 (IL-10) is not only an essential immunoregulator in host immunity, but also it accounts for the intracellular survival of mycobacteria because of its inhibitory activity against anti-mycobacterial functions of macrophage. It has been also indicated that blood cells from calves infected with Mycobacterium avium subsp. paratuberculosis (Map) produce a large amount of IL-10 after stimulation with Map antigen, and it leads to suppression of Interferon-gamma (IFN-gamma) production in T-cells. This characteristic expression of IL-10 in Map infected cattle seems to be playing important roles in the pathogenesis of Johne's disease caused by Map, and could be an important diagnostic indicator. The aim of this study was to investigate the diagnostic significance of IL-10 production from blood cells stimulated by a PPE (Proline-Proline-Glutamic acid) protein family of Map. The recombinant PPE protein, Map41, which has been reported as one of the IFN-gamma inducing antigens of Map, also strongly induced IL-10 from macrophages obtained from infected calves. The elicited IL-10 production in response to Map41 from experimentally infected calves was as early as 2 weeks after the inoculation of Map, and the IL-10 production was detected earlier than that of IFN-gamma. The blood cells from calves immunized with Map produced higher amounts of IL-10 against Map41 stimulation than those of calves immunized with various Mycobacterium species. Furthermore, this IL-10 induction also showed high specificity to Map in guinea pigs experimentally infected with various Mycobacterium species. These observations suggest that IL-10 assay is a useful diagnostic method in the early stage of Johne's disease. PMID- 20018383 TI - Vaccination of ponies with the IE gene of EHV-1 in a recombinant modified live vaccinia vector protects against clinical and virological disease. AB - The control of EHV-1 infection by cytotoxic T-cell responses (CTL) via a reduction in cell associated viremia remains an important goal in horses. Unfortunately, current vaccines are inefficient at inducing these responses. We have identified the immediate early (IE) gene of EHV-1 as a potent stimulator of virus-specific CTL responses in ponies expressing a specific MHC class I serological haplotype (A3/B2). This study was designed to determine if vaccination of A3/B2 MHC I positive ponies with the IE gene could induce protection and immune responses associated with cell mediated immunity. Ponies expressing the MHC-I A3/B2 haplotype (A3/B2 vaccinates) and ponies with a different MHC I haplotype (either non-A3 vaccinates or A3-non-B2 vaccinates) were vaccinated with a recombinant modified vaccinia Ankara (rMVA) vector expressing the IE gene on 3 occasions and vaccinates and unvaccinated controls were challenge infected 8 weeks after the last vaccination. Interferon gamma (IFN gamma) mRNA and antibody titers were determined throughout the study and clinical signs, nasal virus shedding and viremia were determined following challenge infection. Vaccination of A3/B2 vaccinates conferred significant clinical protection and a significant reduction in EHV-1 viremia. IFN-gamma mRNA increased significantly following vaccination in the A3/B2 vaccinates. Antibody titers remained low until after challenge infection, indicating that no accidental field acquired or recrudescent EHV-1 infection had occurred. In summary, this is an important study showing that vaccination of ponies with the EHV-1 IE protein provides not only reduction in clinical disease but also reduction of cell associated viremia, which is a prerequisite for the prevention of abortion and neurological disease. PMID- 20018384 TI - Onset and duration of immunity to equine influenza virus resulting from canarypox vectored (ALVAC) vaccination. AB - Equine influenza virus remains an important problem in horses despite extensive use of vaccination. Efficacy of equine influenza vaccination depends on the onset and duration of protective immunity, and appropriate strain specificity of the immune response. This study was designed to test the protective immunity resulting from vaccination with the North American commercial ALVAC equine influenza vaccine (RECOMBITEK Influenza, Merial, USA)(1) against challenge with American lineage influenza viruses. In experiment 1, 12 ponies were vaccinated twice, at a 35 day interval, using the ALVAC-influenza vaccine expressing the HA genes of influenza A/eq/Newmarket/2/93 and A/eq/Kentucky/94 (H3N8), and 11 ponies served as unvaccinated controls. Six months after the second vaccination, all ponies were challenged with A/eq/Kentucky/91. In experiment 2, 10 ponies received one dose of the ALVAC-influenza vaccine, 10 ponies served as unvaccinated controls, and all ponies were challenge infected with A/equine/Ohio/03, 14 days after vaccination. Parameters studied included serological responses, and clinical disease and nasal viral shedding following challenge infection. In experiment 1, following the two-dose regimen, vaccinated ponies generated high titered anti-influenza virus IgGa and IgGb antibody responses to vaccination and demonstrated statistically significant clinical and virological protection to challenge infection compared to controls. Infection with A/eq/Kentucky/91 produced unusually severe signs in ponies in the control group, requiring therapy with NSAID's and antibiotics, and leading to the euthanasia of one pony. In experiment 2 following the one-dose regimen, vaccinates generated IgGa responses pre-challenge, and anamnestic IgGa and IgGb responses after challenge. Vaccinates demonstrated statistically significant clinical and virological protection to challenge infection compared to controls. The results of this study clearly demonstrate the early onset, and 6-month duration of protective immunity resulting from ALVAC-influenza vaccination against challenge with American lineage equine influenza viruses. PMID- 20018385 TI - Dermoid cyst of the parotid gland: first pediatric case. AB - A dermoid cyst is the result of inclusion of epithelial cells along the lines of embryonic closure. Dermoid cysts of the head neck are uncommon and account for only 7% of all such cysts. They are most often reported as arising in the floor of the mouth. Dermoid cyst is rarely seen in the parotid gland. To our knowledge, there have been only six previous case reports in the English literature. Dermoid cyst of the parotid gland in pediatric patient has not been previously reported in the literature. This is the first case report concerning a dermoid cyst in a pediatric patient. PMID- 20018386 TI - HPV types causing juvenile recurrent laryngeal papillomatosis in South Africa. AB - OBJECTIVES: To identify human papillomavirus (HPV) types associated with juvenile onset recurrent laryngeal papillomatosis (RLP) in southern Africa, to determine if there is a correlation between HPV type and disease aggressiveness and to determine the diagnostic and prognostic value of rapid molecular techniques for detection and typing of HPV using laryngeal biopsies. METHODS: Laryngeal biopsies from patients undergoing surgery for RLP were screened for HPV using conventional and real-time PCR techniques. Amplicons were sequenced to determine the HPV type involved. Clinical features were correlated with HPV type. RESULTS: HPV was identified in papillomata from 18 out of 19 patients. Only HPV-6 and HPV-11 were identified, with no co-infections. There was 100% concordance between conventional and real-time PCR techniques. Patients with HPV-11 disease required more procedures and tended to have higher Derkay scores than those with HPV-6 disease. The HPV types identified in our patients were genetically similar to HPV types from geographically distinct regions. CONCLUSIONS: RLP in our patient population appears to be exclusively due to HPV-6 or HPV-11. HPV-11 disease appears to be more aggressive than HPV-6 disease. Identification of the HPV types provides motivation for inclusion of vaccines against these types in vaccination programs to protect women against infection and subsequently reduce the incidence of RLP. PMID- 20018387 TI - Epiglottitis due to nontypeable Haemophilus influenzae in a vaccinated child. AB - Once a prevalent disease, acute epiglottitis in children has become a rare entity. The introduction of the Haemophilus influenzae type b vaccine has had a dramatic impact on the number of invasive infections caused by this organism. However, physicians must be aware that epiglottitis may result from vaccine failures or from infection with other pathogenic organisms. Vaccinated children with epiglottitis present in a similar fashion to those who are not vaccinated. We present a rare case of acute epiglottitis in a fully vaccinated child due to nontypeable H. influenzae and discuss the clinical presentation and management. PMID- 20018388 TI - A normative study of otoacoustic emissions, ear asymmetry, and gender effect in healthy schoolchildren in Slovakia. AB - OBJECTIVE: The aim of this normative study was to examine cochlear status and possible ear asymmetry and gender effect in transient evoked and distortion product otoacoustic emissions in a group of healthy 12-year-old children in Slovakia. METHODS: Two hundred and twenty-nine 12-year-old children from Slovakia with normal hearing were included in this study. Adolescents with acute infection, abnormal otoscopic findings and abnormal tympanometry were excluded. Pure tone audiometry was performed in standard conditions in a sound proof room. Recordings of transient evoked (TE) and distortion product otoacoustic emissions (DPOAEs) were performed using an ILO 292 USB Echoport. Parameters of hearing thresholds and OAEs were compared using correlation analysis and Wilcoxon test. RESULTS: We found highly statistically significant associations between the hearing thresholds for the left and right ears. When comparing pure tone audiometry with OAEs no significant correlation was found. In TEOAE a significant gender effect and side effect in TEOAE SNR were found. On the other hand there was no side effect in TEOAE response level. In DPOAE neither gender nor side effects were determined. CONCLUSIONS: This is the first comprehensive information on cochlear status among Slovak adolescents. The TEOAEs were significantly higher in girls than boys, but the ear asymmetry in TEOAE was not significant. For DPOAE responses ear asymmetry and gender did not play a role. The data obtained are a basis for population hearing screening, especially for hearing screening programs in infants and children in Slovakia. Moreover data from particular age group represent a link between data from infants and adults. PMID- 20018389 TI - Laryngotracheobronchitis complicated by spontaneous pneumomediastinum. AB - Spontaneous pneumomediastinum (SPM) is an unusual clinical entity that most frequently follows episodes of increased intrathoracic pressures. While typically a benign condition, potentially fatal complications of SPM must be considered and ruled out with each case. We aim to present the first case of croup-associated SPM in the otolaryngology literature and to discuss clinical, diagnostic and management principles. Days following the diagnosis of viral croup, a 7-year-old asthmatic girl presented with unstable vital signs and severe SPM. The patient was urgently taken to the operating room for an endoscopic airway evaluation, which revealed only a mild bacterial croup superinfection. Conservative treatment with inpatient monitoring and antibiotic therapy successfully resolved the episode. SPM should be immediately considered in the differential diagnosis of any patient presenting with cervical emphysema, especially in association with asthma, cough or strenuous activity. Following exclusion of other causes of SPM, conservative and supportive therapies are the mainstays of SPM management. PMID- 20018390 TI - Current status and challenges of antiretroviral research and therapy. AB - Twenty-five years after the discovery of the therapeutic activity of azidothymidine (AZT), the first antiretroviral drug used in the clinic, infection with the human immunodeficiency virus (HIV) has become, at least in the industrialized world, a manageable chronic disease with a significant improvement in life expectancy and quality. Nevertheless, the number of new infections worldwide continues to rise, particularly in women, and effective drug treatments have not yet reached the vast majority of infected individuals in resource limited countries. The current status of antiretroviral therapy is therefore encouraging, but significant challenges remain. Although highly active antiretroviral therapy (HAART) provides durable control of virus replication in many patients, it is not devoid of unwanted secondary effects, some of which are now surfacing in aging populations under long-term treatment. The emergence of multidrug resistance and transmission of drug-resistant HIV strains limit the clinical efficacy of current therapy. Further simplification of treatment and identification of more effective drug combinations are needed to improve patient adherence, the most significant cause of treatment failure. Finding new drugs and novel drug targets may lead to redefining the goals of antiretroviral therapy, with an attempt to achieve the ultimate objective: the eradication of infection. Preclinical and clinical biomedical research, rational drug design and a close collaboration with regulatory agencies to set standards for the transition of new treatment concepts into the clinic will be the cornerstones of future progress. This special issue of Antiviral Research [85(1), 2010] highlights the principal milestones of antiretroviral research over 25 years of drug discovery and development and offers a comprehensive analysis by leading experts of the efforts being made to meet the challenges of effective control of HIV infection. This article forms part of a special issue of Antiviral Research marking the 25th anniversary of antiretroviral drug discovery and development, vol. 85, issue 1, 2010. PMID- 20018392 TI - Assessing the toxicity on [C3mim][Tf2N] to aquatic organisms of different trophic levels. AB - Ionic liquids (ILs) are an exciting class of neoteric solvents that are being object of great attention as a potential replacement to conventional environmental damaging solvents in industrial applications. Despite some progress concerning ILs' toxicity and their environmental impact, the information about these compounds is still scarce. In this work, biological tests were performed to establish the toxicity of 1-methyl-3-propylimidazolium bis(trifluoromethylsulfonyl)imide, [C3mim][Tf2N], in five aquatic species at different trophic levels. Freshwater algal growth inhibition (Pseudokirchneriella subcapitata and Chlorella vulgaris), freshwater cladocerans' immobilization and chronic traits (Daphnia magna and Daphnia longispina) and viability of luminescent marine bacteria (Vibrio fischeri) were investigated. The sensitivity of the different species to the IL was compared in order to determine further repercussions in trophic food web. It is shown that the studied IL is moderately toxic to the studied organisms. P. subcapitata and D. magna are the most tolerant species and C. vulgaris and D. longispina the most sensitive to its presence. PMID- 20018393 TI - Test-and-treat strategy for Helicobacter pylori (HP) infection in older patients. AB - Despite Helicobacter pylori (HP) infection is highly prevalent in older populations, low rates of HP diagnosis and eradication are reported in older persons, even in geriatric wards. We aim to evaluate the results of a HP eradicating program in a sample of older patients in relation to dyspeptic symptoms and to endoscopic findings. A pool of 140 subjects (female/ale=86/54, mean age 68.6+/-5.4 years) with positive C(13)-urea breath test were recruited, clinically evaluated to investigate the presence and characteristics of dyspepsia and abdominal pain, and underwent gastric endoscopic examination. HP-eradication was obtained in 87.9% of subjects with first-line triple therapy, regardless of endoscopic findings. Sixty-eight (48.6%) patients were symptomatic and 72 (51.4%) were asymptomatic. HP-related organic disease in endoscopic examination was frequent (present in 53.6% of the patients) but the distribution of different pathologies were similar in patients with and without symptoms (p=0.86). CONCLUSIONS: even if diagnosis and treatment of HP infection in older patients is underestimated, this population should be a priority for HP-eradication since the infection is more frequent, peptic disease and gastric cancer are frequent, and eradication is effective. PMID- 20018394 TI - Factors associated with impaired urinary albumin excretion in hypertensive individuals: albuminuria and hypertension. AB - Albuminuria is well established as a cardiovascular and renal risk factor. This study aimed to analyze factors associated with this condition in 267 hypertensive individuals, stratified according to urinary albumin excretion (UAE) as: G1--130 individuals with normoalbuminuria (24-h UAE<20 MUg/min); G2--113 with microalbuminuria (UAE between 20 and 200 MUg/min); and G3--24 with macroalbuminuria (UAE>=200 MUg/min). There were significant differences among the groups for blood pressure levels, left ventricular mass and index, estimated glomerular filtration rate, serum creatinine, renal dysfunction and diabetes mellitus (p<0.05). Macroalbuminuria was associated with renal dysfunction (OR=5.91; 95% CI: 2.44-14.31; p<0.001) and microalbuminuria with LVH (OR=2.21; 95% CI: 1.27-3.85; p=0.005). The association between UAE with blood pressure levels, diabetes and target-organ damage in hypertensive individuals suggests that adequate control of cardiovascular risk factors should be pursued to decrease morbidity associated with these conditions. PMID- 20018391 TI - The development of antiretroviral therapy and its impact on the HIV-1/AIDS pandemic. AB - In the last 25 years, HIV-1, the retrovirus responsible for the acquired immunodeficiency syndrome (AIDS), has gone from being an "inherently untreatable" infectious agent to one eminently susceptible to a range of approved therapies. During a five-year period, starting in the mid-1980s, my group at the National Cancer Institute played a role in the discovery and development of the first generation of antiretroviral agents, starting in 1985 with Retrovir (zidovudine, AZT) in a collaboration with scientists at the Burroughs-Wellcome Company (now GlaxoSmithKline). We focused on AZT and related congeners in the dideoxynucleoside family of nucleoside reverse transcriptase inhibitors (NRTIs), taking them from the laboratory to the clinic in response to the pandemic of AIDS, then a terrifying and lethal disease. These drugs proved, above all else, that HIV-1 infection is treatable, and such proof provided momentum for new therapies from many sources, directed at a range of viral targets, at a pace that has rarely if ever been matched in modern drug development. Antiretroviral therapy has brought about a substantial decrease in the death rate due to HIV-1 infection, changing it from a rapidly lethal disease into a chronic manageable condition, compatible with very long survival. This has special implications within the classic boundaries of public health around the world, but at the same time in certain regions may also affect a cycle of economic and civil instability in which HIV-1/AIDS is both cause and consequence. Many challenges remain, including (1) the life-long duration of therapy; (2) the ultimate role of pre exposure prophylaxis (PrEP); (3) the cardiometabolic side-effects or other toxicities of long-term therapy; (4) the emergence of drug-resistance and viral genetic diversity (non-B subtypes); (5) the specter of new cross-species transmissions from established retroviral reservoirs in apes and Old World monkeys; and (6) the continued pace of new HIV-1 infections in many parts of the world. All of these factors make refining current therapies and developing new therapeutic paradigms essential priorities, topics covered in articles within this special issue of Antiviral Research. Fortunately, there are exciting new insights into the biology of HIV-1, its interaction with cellular resistance factors, and novel points of attack for future therapies. Moreover, it is a short journey from basic research to public health benefit around the world. The current science will lead to new therapeutic strategies with far-reaching implications in the HIV-1/AIDS pandemic. This article forms part of a special issue of Antiviral Research marking the 25th anniversary of antiretroviral drug discovery and development, Vol. 85, issue 1, 2010. PMID- 20018396 TI - Budgetary policies and available actions: a generalisation of decision rules for allocation and research decisions. AB - The allocation problem in health care can be characterised as a mathematical programming problem but attempts to incorporate uncertainty in costs and effect have suffered from important limitations. A two-stage stochastic mathematical programming formulation is developed and applied to a numerical example to explore and demonstrate the implications of this more general and comprehensive approach. The solution to the allocation problem for different budgets, budgetary policies, and available actions are then demonstrated. This analysis is used to evaluate different budgetary policies and examine the adequacy of standard decision rules in cost-effectiveness analysis. The research decision is then considered alongside the allocation problem. This more general formulation demonstrates that the value of further research depends on: (i) the budgetary policy in place; (ii) the realisations revealed during the budget period; (iii) remedial actions that may be available; and (iv) variability in parameters values. PMID- 20018395 TI - Risk factors associated with Neospora caninum seropositivity in randomly sampled Canadian dairy cows and herds. AB - Our objective was to determine cow- and herd-level risk factors associated with seropositivity for Neospora caninum in a large number of randomly selected Canadian dairy herds, controlling for important confounding variables and co infections with bovine leukemia virus (BLV), bovine viral diarrhea virus (BVDV) and Mycobacterium avium subspecies paratuberculosis (MAP). Serum samples were obtained from 30 randomly selected cows, where available, in 240 herds using monthly milk testing, within 6 of 10 provinces, and these samples were tested for antibodies against BLV, MAP and N. caninum using commercially available ELISA test kits. Five unvaccinated cattle >6 months old from each herd were tested for antibodies to BVDV using virus neutralization. Most herd-level predictors were obtained through personal interviews with questionnaires administrated to each farm manager. A mixed logistic-regression model was built using N. caninum serostatus at the cow-level as the outcome variable, with herd as a random effect and province as a fixed effect. A BLV seropositive cow was 1.50 times more likely to be seropositive for N. caninum than a BLV-seronegative cow, and this was the only cow-level variable to remain in the final model. Regarding herd-level variables, with "no on-farm dogs" as the baseline, "presence of dogs but not known to eat placentas and/or fetuses" increased the odds of seropositivity for N. caninum by a factor of 1.66. For "presence of dogs known to eat placentas and/or fetuses", the odds ratio (OR) was 2.75, demonstrating a dose-response relationship. "Using embryo transfer" (OR=0.69), "asking for a BVDV-negative test before introducing an animal" (OR=0.30), "using monensin in dry cows" (OR=0.71), and "heifers having nose-to-nose contact with calves" (OR=0.73) were all dichotomous variables negatively associated with seropositivity for N. caninum. "Number of milk cows on the farm" (OR=0.99), and "area (acres) used for forage production" (OR=0.99) were continuous variables negatively associated with N. caninum seropositivity. PMID- 20018397 TI - Pre-soaking of seeds enhances pressure inactivation of E. coli O157:H7 and Salmonella spp. on crimson clover, red clover, radish and broccoli seeds. AB - The application of high hydrostatic pressure (HHP) at a level of 600 MPa at 20 degrees C to decontaminate crimson clover, red clover, radish and broccoli seeds inoculated with E. coli O157:H7 and Salmonella were evaluated. Salmonella was generally more pressure-resistant than E. coli O157:H7 on clover and radish seeds except on broccoli seeds where the trend was reversed. In addition, the application of HHP differentially affected seeds' germinability and the order of pressure tolerance of the seeds was such that red clover>crimson clover approximately broccoli >radish seeds with final germination percentages ranging from 85-100% while their untreated counterparts had final germination percentages of 99-100%. Pre-soaking the different types of seeds in water for 30, 60 or 90 min at ambient temperature followed by HHP at 600 MPa for 2 or 5 min at 20 degrees C significantly (P<0.05) enhanced the pressure inactivation of the inoculated pathogens. Moreover, the ability of HHP-treated seeds to germinate also varied as a function of the pre-soaking duration and the seed type. Pre soaking radish and broccoli seeds for 30 min prior to HHP (2 or 5 min) resulted in germination percentages of 0.05) reduce the sprout yield. The treatment also resulted in a reduction of a 5 log initial load of E. coli O157:H7 and Salmonella to an undetectable level (neither pathogen was detected in 2-g seed samples after enrichment). PMID- 20018398 TI - PTEN mutations and relationship to EGFR, ERBB2, KRAS, and TP53 mutations in non small cell lung cancers. AB - Somatic mutations of phosphatase and tensin homolog deleted on chromosome ten (PTEN) in non-small cell lung cancers (NSCLCs) have been investigated in but a small number of cases. In addition, the relationship between PTEN mutations and epidermal growth factor receptor (EGFR), KRAS, and TP53 mutations has not been investigated. Therefore, we investigated the frequency of PTEN mutations in 176 surgically resected NSCLCs and analyzed the relationship between PTEN mutations and EGFR, ERBB2, KRAS, and TP53 mutations. Mutations of PTEN (exons 1-9), EGFR (exons 18-21), ERBB2 (exons 19 and 20), KRAS (exon 1), and TP53 (exons 2-11) were determined by polymerase chain reaction and direct sequencing. PTEN mutations were present in 8 (4.5%) of the 176 tumors. PTEN mutations were only found in ever-smokers and were significantly more frequent in squamous cell carcinoma than in adenocarcinoma (10.2% vs 1.7%, P=0.02). Mutations of EGFR, ERBB2, KRAS, and TP53 genes were found in 36 (20.5%), 2 (1.1%), 11 (6.3%), and 66 (37.5%) cases, respectively. Of the 8 tumors with PTEN mutations, 1 case concurrently had an EGFR mutation and 4 cases had TP53 mutations. However, PTEN mutations were not found in the tumors with KRAS mutation. Our findings indicate that PTEN mutations are relatively common in NSCLC, and thus analysis of PTEN mutations may facilitate a comprehensive understanding of the genetic alterations related to the EGFR signaling pathway. PMID- 20018399 TI - Investigation and management of lung cancer in older adults. AB - BACKGROUND: The average age of patients with lung cancer is increasing, but there is little data of the management and outcomes of this common cancer in the elderly. METHODS: Data were collected from our hospital lung cancer database, medical, radiology and pathology records, for all patients with a new diagnosis of lung cancer from Jan 2002 to Dec 2004. The investigation, treatment and outcomes of these patients were analysed, and the younger (<75 years old) and elderly (75 years+) populations were compared. RESULTS: Data on 367 were analysed. Median age at diagnosis was 72 years and median survival was 5.2 months. Elderly patients were less likely to be fully investigated i.e. have histological confirmation (88.2% vs. 66.4%; p<0.001), stage recorded (88.6% vs. 78.1%; p=0.001) and performance status recorded (88.6% vs. 75.3%; p=0.003). They had poorer performance status. The older patients were less likely to receive active treatment (surgery 11.6% vs. 6.4%; p<0.001, chemotherapy or radiotherapy 50.9% vs. 26.2% p<0.001). Improved survival was independently associated with younger age (p<0.001), better performance status (p<0.001), early stage (p<0.001) and active treatment (p=0.005). Hazard ratios for death for poor PS vs. good PS was 1.88 (p<0.001), for late stage vs. early stage was 2.01 (p=0.011), for BSC vs. active treatment was 1.46 (p=0.005) and for 75 years+ patients vs. under 75 years was 1.36 (p=0.069). When comparing elderly with younger patients, median survival remained better in patients who were actively treated, had good PS and early stage (p<0.001). CONCLUSIONS: Elderly patients are less likely to be fully investigated and actively treated than younger patients with lung cancer. The causes of this difference are unclear but may include patient and clinicians' more conservative approach in the elderly. Predictors of improved outcome are the same in older and younger populations, and the elderly derive a similar survival advantage if actively treated. These data show that elderly patients are managed differently to younger patients, though the clinical basis for this is not clear. SUMMARY: Lung cancer is common in the older population and has a poor prognosis. We reviewed the investigation, treatment and survival of all patients with a new diagnosis of lung cancer over a three-year period. We compared the management of our younger (under 75 years) population with those aged 75 years or older. In our population, people over 75 years with lung cancer are less completely investigated. They were less likely to have full staging investigations, a histological or cytological diagnosis, or have their performance status recorded. Our over 75-year olds are less likely to receive active treatment (chemotherapy, radiotherapy or surgery) for lung cancer. Our older patients who received active treatment for lung cancer had similar survival advantages to younger patients. PMID- 20018400 TI - Analyzing female gametophyte development and function: There is more than one way to crack an egg. AB - In flowering plants, gametes are formed in specialized haploid structures, termed gametophytes. The female gametophyte is a few-celled structure that integrates such diverse functions as pollen tube attraction, sperm cell release, gamete fusion and seed initiation. These processes are realized by distinct cell types, which ensure reproductive success in a coordinated manner. In the past decade, much progress has been made concerning the molecular nature of the functions carried out by the different cell types. Here, we review recent work that has shed light on female gametophyte development and function with a particular focus on approaches that have led to the isolation of genes involved in these processes. PMID- 20018401 TI - Role of A-type ARABIDOPSIS RESPONSE REGULATORS in meristem maintenance and regeneration. AB - In Arabidopsis, the network responsible for the maintenance of the shoot apical meristem (SAM) is built on a negative feedback loop involving the peptide ligand CLAVATA3 (CLV3) and the homeodomain transcription factor WUSCHEL (WUS). The local WUS/CLV3 regulatory module is linked to the organism-wide cytokinin signalling system by direct transcriptional control of A-type ARABIDOPSIS RESPONSE REGULATOR genes (ARRs) by WUS. Here we investigate two A-type ARR genes, ARR7 and ARR15, which are negative regulators of cytokinin signalling. We show that the expression of ARR7, WUS and CLV3 is dependent on cytokinin signalling. While ARR7 expression strongly responds to variations in cytokinin activity, WUS and CLV3 appeared to be much more buffered against this type of variation. As earlier studies had shown that pertubation of A-type ARR activity only causes mild effects on SAM function, we employed tissue regeneration assays as a sensitised background. Root explants pre-treated on auxin and cytokinin-rich callus-inducing medium showed severely suppressed shoot regeneration when ARR7 and ARR15 were overexpressed, whereas loss of function of these genes had a strongly promoting effect. This phenotype was even aggravated in the arr3,4,5,6,7,8,9 septuple mutant. Futhermore, loss-of A-type ARR function in arr7 and arr3,4,5,6,7,8,9 mutants strongly stimulated callus development, indicating that cell proliferation is repressed by A-type ARRs. To elucidate the mechanisms underlying the enhanced capacity of the arr3,4,5,6,7,8,9 septuple mutant to develop shoot tissue in culture, we used whole-genome expression profiling. Among the transcripts with increased abundance in arr3,4,5,6,7,8,9 inflorescence apices a strong enrichment for functions in pollen development was apparent, while the reduced transcripts showed a more heterogeneous distribution of functional categories, ranging from development to pathogen defence. PMID- 20018402 TI - The multifunctional leucine-rich repeat receptor kinase BAK1 is implicated in Arabidopsis development and immunity. AB - Plant receptor-like kinases (RLKs) are transmembrane proteins with putative N terminal extracellular ligand-binding domains and C-terminal intracellular protein kinase domains. RLKs have been implicated in multiple physiological programs including plant development and immunity to microbial infection. Arabidopsis thaliana gene expression patterns support an important role of this class of proteins in biotic stress adaptation. Here, we provide a comprehensive survey of plant immunity-related RLK gene expression. We further document the role of the Arabidopsis Brassinosteroid Insensitive 1 (BRI1)-associated receptor kinase 1 (BAK1) in seemingly unrelated biological processes, such as plant development and immunity, and propose a role of this protein as an adaptor molecule that is required for proper functionality of numerous RLKs. This view is supported by the identification of an additional RLK, PEPR1, and its closest homolog, PEPR2 as BAK1-interacting RLKs. PMID- 20018403 TI - Cellular analysis of newly identified Hox downstream genes in Drosophila. AB - Hox genes code for conserved homeodomain transcription factors, which act as regional regulators for the specification of segmental identities along the anterior-posterior axis in all animals studied. They execute their function mainly through the activation or repression of their downstream genes. We have recently identified a large number of genes to be directly or indirectly targeted by Hox proteins through gene expression profiling in the model organism Drosophila. However, the cell-specific regulation of these downstream genes and the functional significance of the regulation are largely unknown. We have validated and functionally studied many of the newly identified downstream genes of the Hox proteins Deformed (Dfd) and Abdominal-B (Abd-B), and provide evidence that Hox proteins regulate a diverse group of downstream genes, from transcription factors to realisators with major and minor roles during morphogenesis. PMID- 20018404 TI - Allelopathy and allelopathic substance in the moss Rhynchostegium pallidifolium. AB - An aqueous methanol extract of the moss Rhynchostegium pallidifolium, which often forms large pure colonies on soils and rocks, inhibited the growth of cress (Lepidium sativum), alfalfa (Medicaga sativa), lettuce (Lepidium sativum), ryegrass (Lolium multiflorum), timothy (Phleum pratense) and Digitaria sanguinalis seedlings. Increasing the extract concentration increased the inhibition. These results suggest that R. pallidifolium may show allelopathic activity. The extract was purified and a putative compound causing this growth inhibitory effect was isolated. The chemical structure of the growth inhibitor was determined by MS, and (1)H and (13)C NMR spectral data as 3-hydroxy-beta ionone. 3-Hydroxy-beta-ionone inhibited the shoot and root growth of cress seedlings at concentrations greater than 1 and 3 micromol/L, respectively. The doses required for 50% growth inhibition on the shoot and roots of cress seedlings were 16.3 and 14.9 micromol/L, respectively. The endogenous concentration of 3-hydroxy-beta-ionone in R. pallidifolium was 28.2 microg/g and the concentration of 3-hydroxy-beta-ionone in the growth medium of R. pallidifolium was 6.7 microg/g. These results suggest that 3-hydroxy-beta-ionone was likely secreted into the medium during the incubation of R. pallidifolium. In addition, 3-hydroxy-beta-ionone was found in the soil under the pure colonies of R. pallidifolium. Therefore, 3-hydroxy-beta-ionone may play an important role in the allelopathic activity of R. pallidifolium and may help competition with neighboring plants, resulting in the formation of pure colonies. PMID- 20018405 TI - A population-based study of cutaneous melanoma in Alberta, Canada (1993-2002). AB - BACKGROUND: There is evidence to suggest that melanoma incidence rates continue to rise in Canada and the United States. OBJECTIVE: Our objective was to determine cutaneous melanoma trends from 1993 to 2002 in the province of Alberta and to compare the results to previously published provincial analyses for the decade of 1967-1976. METHODS: A retrospective study of 3479 patients with cutaneous melanoma diagnosed in Alberta between 1993 and 2002 was conducted. Estimates of relative survival compared the survival of melanoma patients with the Alberta population to derive the likelihood of surviving melanoma in the absence of other causes of death. Further comparison to published Canadian data was also conducted. RESULTS: For the period 1993-2002, the annual melanoma age standardized incidence rates per 100,000 person-years ranged between 11.1 and 15.9 and between 9.8 and 14.1 among men and women, respectively. These rates are considerably higher than the previously reported (1976) highest Alberta incidence rates of 4.1 and 4.8 in men and women, respectively. The rates increased slightly for the period 1993-1999 with an average annual percentage change of +3.5%, but appeared to decrease for the interval 1999-2002 with an average annual percentage change of -6.4%. The majority of the tumors were less than 1.0 mm in thickness for both genders. On univariate analysis the following parameters were associated with decreasing patient survival: male gender, increasing age, head and neck tumors, Clark level of invasion, and Breslow tumor thickness. Multivariate analysis demonstrated that the strongest determinant of survival was Breslow tumor thickness. LIMITATIONS: Melanomas in-situ were not included in this study. CONCLUSIONS: Although melanoma incidence rates in Alberta are higher than previously reported, the incidence rates over the study period of 1993 to 2002 appear to have leveled and may in fact be declining over the past several years. PMID- 20018406 TI - Low rates of clinical recurrence after biopsy of benign to moderately dysplastic melanocytic nevi. AB - BACKGROUND: Little is known about the recurrence/persistence rates of dysplastic nevi (DN) after biopsy, and whether incompletely removed DN should be re-excised to prevent recurrence. OBJECTIVE: Our purpose was to determine the recurrence rates of previously biopsied DN, and to assess whether biopsy method, margin involvement, congenital features, epidermal location, and degree of dysplasia are associated with recurrence. METHODS: Patients having a history of a "nevus biopsy" at least 2 years earlier were assessed for clinical recurrence. Slides of original lesions were re-reviewed by a dermatopathologist. RESULTS: A total of 271 nevus biopsy sites were assessed in 115 patients. Of 195 DN with greater than 2 years of follow-up, 7 (3.6%) demonstrated recurrence on clinical examination. In all, 98 DN had a follow-up period of at least 4 years with no clinical recurrence. Of 61 benign nevus biopsy sites examined, clinical recurrence was observed in two (3.3%). For all nevi, recurrence was significantly associated with shave biopsy technique but not with nevus dysplasia or subtype, or the presence of positive margin or congenital features. LIMITATIONS: Most biopsies were performed in a pigmented lesion clinic at a single tertiary referral center. Determinations of nevus recurrence were made on clinical rather than histologic grounds, and follow-up times were limited in some cases. CONCLUSION: In this cohort, rates of clinical recurrence after biopsy of DN and benign nevi were extremely low. Re-excision of nevi, including mildly to moderately DN with a positive margin, may not be necessary. PMID- 20018407 TI - C-terminal FUS/TLS mutations in familial and sporadic ALS in Germany. AB - Amyotrophic lateral sclerosis (ALS), the major form of motor neuron disease in the adult occurs as a sporadic disease in more than 95% of all cases. Analysis of familial forms is considered as a key to understand the pathophysiology of the disease. It is expected that mutations responsible for familial forms are also found in sporadic ALS. During the past years, several loci and genes have been identified in which disease associated mutations have been discovered. We report here on the screening of 596 sporadic ALS patients, 41 familial ALS cases and other motor neuron disease patients from Germany for mutations in the FUS/TLS gene. Sequencing of the last two exons in all patients revealed the C1561T transversion, which leads to the amino acid substitution at R521C, in one familial and one sporadic ALS patient. In addition three patients with a synonymous mutation at codon 522 were identified. None of these variants were present in the control population. Our results indicate that mutations in FUS/TLS are not a major cause of sporadic ALS in the German population. PMID- 20018408 TI - Fractalkine and CX 3 CR1 regulate hippocampal neurogenesis in adult and aged rats. AB - Microglia have neuroprotective capacities, yet chronic activation can promote neurotoxic inflammation. Neuronal fractalkine (FKN), acting on CX(3)CR1, has been shown to suppress excessive microglia activation. We found that disruption in FKN/CX(3)CR1 signaling in young adult rodents decreased survival and proliferation of neural progenitor cells through IL-1beta. Aged rats were found to have decreased levels of hippocampal FKN protein; moreover, interruption of CX(3)CR1 function in these animals did not affect neurogenesis. The age-related loss of FKN could be restored by exogenous FKN reversing the age-related decrease in hippocampal neurogenesis. There were no measureable changes in young animals by the addition of exogenous FKN. The results suggest that FKN/CX(3)CR1 signaling has a regulatory role in modulating hippocampal neurogenesis via mechanisms that involve indirect modification of the niche environment. As elevated neuroinflammation is associated with many age-related neurodegenerative diseases, enhancing FKN/CX(3)CR1 interactions could provide an alternative therapeutic approach to slow age-related neurodegeneration. PMID- 20018409 TI - LRRK2 variant associated with Alzheimer's disease. AB - Overlapping neurodegenerative pathologies (including Alzheimer's disease, AD) have been described in Parkinson's disease (PD) patients with leucine-rich repeat kinase-2 (LRRK2) mutations. We analyzed a LRRK2 PD (R1628P) risk variant in a group of 885 subjects comprising of AD and controls. The frequency of the R1628P allele was higher in AD compared to controls (3.5% vs. 1.6%, OR 2.3, 95 CI 1.2 4.4, p=0.018). In vitro, the mean percentage of apoptosis and cell death observed for the R1628P transfected human cell lines was higher compared to wild type 21.8 +/- 1.9, vs. 17.1 +/- 1.3, p<0.05, 30.2 +/- 2.2 vs. 25.7 +/- 1.3, p<0.05). The LRRK2 R1628P variant increases the risk of AD in our population and our in vitro findings suggest that it is a functional variant and predisposes to apoptosis. PMID- 20018410 TI - Chronic systemic treatment with a high-dose proteasome inhibitor in mice produces akinesia unrelated to nigrostriatal degeneration. AB - Supporting the hypothesis that proteasome dysfunction is involved in Parkinson's disease (PD), McNaught et al. (2004) reported that the systemic administration of the proteasome inhibitor Z-Ile-Glu(OtBu)-Ala-Leu-aldehyde (PSI) in rats led to the degeneration of the nigrostriatal pathway. However, several groups could not reproduce this finding. We herein attempted to improve the reliability of the PSI model by chronically delivering the inhibitor using osmotic minipumps in aged mice. We also tested whether PSI co-administered with 1-methyl-4-phenyl-1,2,3,6 tetrahydropyridine (MPTP) could act synergistically to induce toxicity. We found that PSI produced a significant reduction in locomotor activity that was mildly exacerbated by MPTP. However, PSI alone produced no sign of degeneration of the nigrostriatal dopaminergic pathway and did not exacerbate MPTP toxicity. To conclude, PSI administration does not provide a reliable phenotypic model of PD. PMID- 20018412 TI - [Treatment with (131)I of thyroid remnants in a patient with papillary thyroid carcinoma and end-stage chronic renal failure]. AB - The follow-up and treatment of thyroid cancer presents several aspects subject to discussion, such as its management in patients with End-Stage Renal Failure (ESRF). We present a patient with ESRF and papillary thyroid carcinoma, which had to be coordinated among different departments (Endocrinology, Nuclear Medicine, Nephrology and Physics and Radiation Protection). Both the diagnostic scintigraphy with (123)I and the ablative treatment with (131)I performed later were performed with the administration of rh TSH. The room in which the metabolic therapy was to be performed was prepared for the patient's periodic hemodialysis. The (131)I dose used was 80% of the usual dose. This made it possible to assure the therapeutic effect and that the patient's stay in hospital would only be for 5 days. Throughout the whole diagnostic and therapeutic process, no adverse effects attributable to rh TSH or radioiodine were observed. The coordination among the departments involved enabled an effective and safe process for the patient. PMID- 20018411 TI - Age-related deficits in a forebrain-dependent task, trace-eyeblink conditioning. AB - Trace-eyeblink conditioning is a forebrain-dependent learning paradigm that has assisted in our understanding of age-related hippocampal neuronal plasticity; however, the hippocampus is not believed to be the permanent site for most long term-memory storage. Studies in adult subjects have suggested the neocortex as one such site. Whisker plucking studies have further suggested that the ability for plasticity in the neocortex declines with age. Mice were trained in trace- and delay-eyeblink conditioning with whisker or auditory stimulation as the conditioned stimulus to examine possible age-related behavioral and neocortical abnormalities. Whisker stimulation was determined to be a more effective stimulus for examining age-related behavioral abnormalities in C57 mice. Additionally, neocortical barrel expansion, observed in trace conditioned adult mice and rabbits, does not occur in mice conditioned on a delay paradigm or in old mice unable to learn the whisker trace association. Abnormalities in neocortical memory storage in the elderly could contribute to normal age-dependent declines in associative learning abilities. PMID- 20018413 TI - Pneumothorax after orthognathic surgery. AB - We report the case of an 18-year-old girl who had pneumothorax two days after bimaxillary osteotomy to correct a severe Class II malocclusion. Chest radiograph showed left-sided pneumothorax with complete collapse of the underlying lung and mediastinal shift to the left. A chest drain was inserted and she made an uneventful recovery. The aetiology may be iatrogenic or spontaneous, but we have found no previous reports of pneumothorax after orthognathic surgery. PMID- 20018414 TI - A case of Collet-Sicard syndrome presenting to the Oral and Maxillofacial Surgery Department and a review of the literature. AB - A case of Collet-Sicard Syndrome caused by skull base metastasis of probable breast adenocarcinoma is reported. A 79-year-old lady presented to the Oral and Maxillofacial Surgery Department with progressive left-sided tongue symptoms that she described as swelling. This was initially attributed to local disease, and a biopsy showed the patient was suffering from Necrotizing Sialometaplasia. However, her symptoms rapidly evolved into cranial nerve palsies affecting IX XII, not initially diagnosed. Subsequent imaging revealed the cause of her worsening symptoms to be a metastatic lesion at her left skull base. Cranial nerve palsies due to metastases to the skull base are rare, and the authors would advise clinicians to adopt a high-index of suspicion in ruling out cranial nerve pathology at the skull base when encountering unusual signs and symptoms in the head and neck region. PMID- 20018415 TI - Perforation of the inferior alveolar nerve by the maxillary artery: an anatomical study. AB - The infratemporal fossa is a clinically important anatomical area for the delivery of local anaesthetic agents in dentistry and maxillofacial surgery. We studied the infratemporal fossas in white cadavers, and in particular the topographical relations of the inferior alveolar nerve and the maxillary artery. In 3 of the 50 fossas dissected the maxillary artery passed through the inferior alveolar nerve, splitting it into superficial and deep divisions. Entrapment of the maxillary artery may cause numbness or headache and may interfere with injection of local anaesthetics into the infratemporal fossa. PMID- 20018416 TI - Acceleration of consolidation period by thrombin peptide 508 in tibial distraction osteogenesis in rats. AB - We aimed to find out whether a single local injection of two different doses of thrombin peptide 508 (TP508) could accelerate consolidation of bone in sites of tibial distraction osteogenesis in rats. Forty-eight adult male Sprague-Dawley rats were divided equally (n=16 in each group) into controls (given saline alone), and two experimental groups, given injections of TP508 10MUg or 100 MUg. The animals were killed on days 14 and 28 after distraction. Histomorphometric evaluation showed that the TP508 resulted in significantly larger areas of newly formed bone (p<0.003 and p<0.0001) than saline alone. At 2 weeks, more new bone had formed in the group given TP508 100 MUg than in the group given 10 MUg, but the difference was not significant (p=0.8). However, the difference was significant at 4 weeks (p=0.03). These findings suggest that a single injection of TP508 given at the end of the distraction period increased the degree of consolidation. The higher dose was more effective at the later time point. PMID- 20018417 TI - Maxillary aseptic necrosis after Le Fort I osteotomy: a case report and literature review. PMID- 20018418 TI - Bioabsorbable plates and screws for fixation of mandibulotomies in ablative oral cancer surgery. AB - PURPOSE: The use of bioabsorbable self-reinforced poly(L/DL)lactide 70/30 (SR PLDLA) miniplates and screws could offer many benefits compared with conventional metallic devices for fixation of access osteotomies in oral cancer surgery. The material neither interferes with imaging nor with postoperative radiotherapy. The plates and screws do not need to be removed. The number of operations can be diminished and rehabilitation with dental implants shortened. The purpose of the present study was to report our experience with bioabsorbable osteosynthesis in the fixation of access osteotomies of the mandible. PATIENTS AND METHODS: A total of 15 patients (11 men and 4 women, mean age 63.1 years) were enrolled in the present study. All patients had oral squamous cell carcinoma. Three patients developed tumor recurrence, 2 of whom had previously undergone radiotherapy. Nine patients had radiotherapy scheduled postoperatively. The osteotomies were a straight-line cut and were situated medially or paramedially. For fixation, SR PLDLA miniplates and screws (Biosorb 2.0 and 2.4 systems) were used without any maxillomandibular fixation. RESULTS: The follow-up ranged from 0.3 to 7.1 years (median 3.5). No problems were encountered during the operation. One patient required reoperation owing to failure in fixation. Twelve osteotomy lines (80%) were clinically stable; radiologically, 6 were totally and 3 partly consolidated. During follow-up, 6 nonunions were radiographically noted, 3 of which were clinically stable. CONCLUSIONS: Because of the high incidence of radiologic nonunion, bioabsorbable devices should not yet be used for fixation of access osteotomies in cancer surgery. PMID- 20018419 TI - Facial artery pseudoaneurysm following surgical removal of a mandibular molar. PMID- 20018420 TI - The role of cytoreductive surgery for non-genital tract metastatic tumors to the ovaries. AB - OBJECTIVE: The aim of this study was to investigate prognostic factors of patients with metastases to the ovaries from non-genital organs. STUDY DESIGN: From September 1994 to December 2006, 158 patients with pathologically confirmed metastatic tumors to the ovaries at Samsung Medical Center (SMC) were included in this study. The data were obtained from the patients' medical records and pathology reports. RESULTS: The primary tumor origin was mostly stomach (73 cases) and colon (61 cases). Krukenberg tumor (pathologically proven signet ring cell carcinoma) was found in 34 cases: stomach (25), colon (2), appendix (1), and unknown (6). Residual disease after surgery was >2 cm in 65 (41.1%) cases and <2 cm in 93 (58.9%) cases. The overall 5-year survival was 7.2% and the median survival time was 15 months. The median survival times according to the primary tumor site showed significant differences (p=0.002) and were as follows: stomach 12 months, colon 17 months. The median survival in cases with residual disease <2 cm vs. >2 cm was 26 months vs. 15 months (p=0.017) and the median survival with vs. without adjuvant chemotherapy was 16 months vs. 10 months (p=0.001). However, age, bilateral tumors, chronology of diagnosis and mass size did not affect survival. CONCLUSION: Cytoreductive surgery and post-operative adjuvant chemotherapy had a beneficial effect on survival in selected patients. PMID- 20018421 TI - A randomised controlled trial of placental cord drainage to reduce feto-maternal transfusion. AB - OBJECTIVES: To determine whether placental drainage via the umbilical cord prior to placental delivery reduces the size of feto-maternal transfusion and thus the chance of rhesus isoimmunisation in rhesus negative women. STUDY DESIGN: A randomised controlled trial conducted in a tertiary hospital setting in the UK compared 18 rhesus negative women who had placental drainage (10 caesarean section and 8 vaginal deliveries) with 18 rhesus negative women where the cord remained clamped until placental delivery (8 caesarean section and 10 vaginal deliveries). Maternal venous blood samples were taken before delivery and at a mean of 142 min after delivery of the placenta, and analysed using flow cytometry to calculate the size of the feto-maternal transfusion. The statistical analysis was performed using SPSS Version 13 statistical software. The main outcome measure was the quantification of the volume of fetal cells in the maternal circulation before and after delivery. RESULTS: In the 72 specimens taken, 40 demonstrated measurable amounts of fetal cells in the maternal circulation. In the 18 women who had placental drainage, the mean (SD) size of the feto-maternal transfusion was 0.50 ml (0.79) before and 0.39 ml (0.58) after delivery. In the 18 women who had a clamped cord, the mean (SD) feto-maternal transfusion was 0.46 ml (0.84) before and 0.78 ml (1.1) after delivery. There was no significant difference between the net feto-maternal transfusions in the two groups (Mann Whitney U 122.5, p 0.19). CONCLUSION: Placental drainage does not reduce the amount of feto-maternal transfusion and this method of placental delivery is not recommended to reduce feto-maternal transfusion. PMID- 20018422 TI - Different physical treatment modalities for lymphoedema developing after axillary lymph node dissection for breast cancer: a review. AB - The purpose of this article is to provide a systematic review of Combined Physical Therapy, Intermittent Pneumatic Compression and arm elevation for the treatment of lymphoedema secondary to an axillary dissection for breast cancer. Combined Physical Therapy starts with an intensive phase consisting of skin care, Manual Lymphatic Drainage, exercises and bandaging and continues with a maintenance phase consisting of skin care, exercises, wearing a compression sleeve and Manual Lymphatic Drainage if needed. We have searched the following databases: PubMed/MEDLINE, CINAHL, EMBASE, PEDro and Cochrane. Only (pseudo-) randomised controlled trials and non-randomised experimental trials investigating the effectiveness of Combined Physical Therapy and its different parts, of Intermittent Pneumatic Compression and of arm elevation were included. These physical treatments had to be applied to patients with arm lymphoedema which developed after axillary dissection for breast cancer. Ten randomised controlled trials, one pseudo-randomised controlled trial and four non-randomised experimental trials were found and analysed. Combined Physical Therapy can be considered as an effective treatment modality for lymphoedema. Bandaging the arm is effective, whether its effectiveness is investigated on a heterogeneous group consisting of patients with upper and lower limb lymphoedema from different causes. There is no consensus on the effectiveness of Manual Lymphatic Drainage. The effectiveness of skin care, exercises, wearing a compression sleeve and arm elevation is not investigated by a controlled trial. Intermittent Pneumatic Compression is effective, but once the treatment is interrupted, the lymphoedema volume increases. In conclusion, Combined Physical Therapy is an effective therapy for lymphoedema. However, the effectiveness of its different components remains uncertain. Furthermore, high-quality studies are warranted. The long-term effect of Intermittent Pneumatic Compression and the effect of elevation on lymphoedema are not yet proven. PMID- 20018423 TI - A cutaneous-vaginal fistula and myositis of the obturator muscle following placement of a trans-obturator tape for stress incontinence. PMID- 20018424 TI - Re-vascularisation in human ovarian tissue after conventional freezing or vitrification and xenotransplantation. AB - OBJECTIVE: In ovarian tissue grafts there is a massive loss of follicles during the ischaemic period until re-vascularisation is established. The aim of our study was to investigate the influence of different cryopreservation techniques on the ability for the re-vascularisation of ovarian tissue transplanted to SCID mice. STUDY DESIGN: Ovarian fragments from five patients were cut into pieces (approximately 0.5 mm x 1.0 mm x 1.0 mm) and randomly distributed into three groups: fresh non-treated tissue (group A); tissue conventionally frozen in standard 0.5 ml insemination straws with 1.5 M ethylene glycol+0.1 M sucrose, with thawing in a 40 degrees C water bath and step-wise removal of cryoprotectants at room temperature in 0.5 M, 0.25 M and 0.15 M sucrose with gentle agitation (group B); tissue vitrified in 2.62 M dimethylsulphoxide+2.6 M acetamide+1.31 M propylene glycol+0.0075 M polyethylene glycol, with warming by direct plunging of solid specimens with ovarian pieces into 20 ml of 50% vitrification solution pre-warmed to 40 degrees C and dilution of cryoprotectants in a decreasing concentration of vitrification solution (25%, 12.5%) at room temperature (group C). We used a xenograft model in which ovarian tissue pieces of all three groups were subcutaneously transplanted in SCID mice. The animals were sacrificed on the third day after ovarian tissue transplantation and then weekly during 1 month to obtain the ovarian tissue grafts. These samples were examined by immunohistochemical staining with the endothelial cell-specific marker platelet endothelial cell adhesion molecule-1 (PECAM-1) to determine angiogenesis. Histological observation of tissue after explantation was performed and quality and quantity of follicles were assessed. RESULTS: No PECAM-1 staining was observed in all treatment groups prior to grafting. After warming and in vivo culture of ovarian tissue, the beginning of angiogenesis in pieces from all treatment groups on the third day was detected by PECAM-1 staining. After 4 weeks of in vivo culture the overall area of PECAM-1-positive blood vessels significantly increased (P<0.05), independent of the type of cryopreservation (groups B and C vs. group A). It was found that transplantation technique had negative influence on the integrity of follicles independent of the type of treatment during in vivo culture. The duration of in vivo culture has a negative, but not statistically significant, influence on follicle quality in long-cultured transplants inside each treatment group (P>0.5). CONCLUSION: The process of re vascularisation of transplanted ovarian tissue is independent of the type of treatment and does not influence follicle quality. PMID- 20018425 TI - Efficacy of inside-out transobturator vaginal tape (TVTO) at 4 years follow up. AB - OBJECTIVES: To assess the efficacy and complications of TVTO with or without anterior colporrhaphy in women with urodynamic stress incontinence at 4 years follow up. STUDY DESIGN: Seventy-four patients were subjected to TVTO operation and 41 patients to TVTO and anterior colporrhaphy. Subjective and objective cure and improvement rates and the incidence of complications were assessed. RESULTS: The objective cure rate based on the pad test findings for the TVTO-only patients was 82.4% and the improvement rate was 6.8%. The objective cure rate for the group undergoing TVTO and anterior colporrhaphy was 80.5% and the improvement rate was 7.4%. CONCLUSIONS: At 4 years follow up TVTO procedure alone or with anterior colporrhaphy maintains a high cure and improvement rate with a very low complication rate and appears to be a promising technique, but long term results should be published for safer conclusions to be made. PMID- 20018426 TI - GnRH-II mRNA expression in tumor tissue and peripheral blood mononuclear cells (PBMCs) in patients with malignant and benign ovarian tumors. AB - OBJECTIVE: To investigate the expression of the second form of GnRH (GnRH-II) in tumor tissue and peripheral blood mononuclear cells (PBMCs) in malignant and benign ovarian tumors in humans. STUDY DESIGN: Sixty-six women were studied: 24 with epithelial ovarian carcinomas, 22 with benign ovarian tumors and 20 in the control group undergoing surgery. Malignant, benign and normal ovarian tissue and PBMCs were obtained for measurement of GnRH-II mRNA levels using quantitative real-time RT-PCR. RESULT(S): The expression of GnRH-II was found to be 1.5 times higher in malignant ovarian tumors compared with benign ovarian tumors and the control group in post-menopausal patients (P<0.01). In the post-menopausal patient group with malignant ovarian tumors, there were significant positive correlations between serum FSH level and ovarian tissue GnRH-II mRNA expression (r=0.68; P=0.03), and serum LH level and ovarian tissue GnRH-II mRNA expression (r=0.71; P=0.02). Controls, benign and malignant groups were similar in terms of GnRH-II expression in PBMCs in the pre- and post-menopausal periods. There was no significant correlation between ovarian tissue GnRH-II mRNA expression vs. PBMC GnRH-II mRNA expression in patient and control groups. CONCLUSION(S): We have shown increased GnRH-II expression in human ovarian cancer tissue in post menopausal women in vivo. Expression of GnRH-II in PBMCs did not reflect the local GnRH-II expression levels in ovarian tissue. These preliminary data suggest that local GnRH-II may participate in the regulation of ovarian tumor growth in post-menopausal women. PMID- 20018427 TI - Dealing with pollution from conflict: Analysis of discourses around the 2006 Lebanon oil spill. AB - In July 2006 a war between Lebanon and Israel resulted in severe environmental damage in Lebanon from Israeli bombing raids. An attack on the Lebanese Jiyyeh Power Plant released 15,000 tons of heavy fuel oil into the Mediterranean Sea. Remarkably, a clean-up operation was effected despite a continued state of war and lack of capacity in the Lebanese government. Civil society environmentalists played a key role in dealing with the pollution and complying with pollution control legislation. In this study we use Q-methodology to analyse discourses on the effectiveness of pollution legislation during times of conflict using the Jiyyeh oil spill as an example. We interviewed 35 people from eight different stakeholder groups involved in environmental issues. Five distinct discourses were generated covering compensation schemes, need for new legislation, role of stakeholders during wartime and strengthening government ministries. PMID- 20018428 TI - Hypocrellin B enhances ultrasound-induced cell death of nasopharyngeal carcinoma cells. AB - Hypocrellin B, a natural pigment from a traditional Chinese herb, has been attracting extensive attention. The present study aims to investigate whether hypocrellin B can enhance cell death induced by ultrasound sonification on nasopharyngeal carcinoma cells in vitro. The sonodynamic action of hypocrellin B was investigated on nasopharyngeal carcinoma cell line CNE2 cells as tumor model cells. In the experiments, the hypocrellin B concentration was kept constant at 2.5 microM and the cells were subject to ultrasound exposure for 15 s at an intensity of 0.65 W/cm(2). Cytotoxicity was investigated 24 h after ultrasound sonification. Apoptosis was evaluated using flow cytometry with annexin V-FITC and propidium iodine staining and nuclear staining with Hoechst 33258. Cell ultrastructure morphology was observed using transmission electron microscopy (TEM). No significant dark cytotoxicity of hypocrellin B in the CNE2 cells was observed at the concentration of 2.5 microM. The cell death rate induced by ultrasound sonification was significantly higher in the presence of hypocrellin B than in the absence of hypocrellin B. Flow cytometry showed that ultrasound exposure in the presence of hypocrellin B significantly increased the early and late apoptotic rate, 18.64% and 22.57%, respectively, compared with the controls. Nuclear condensation was observed in the nuclear staining and swollen mitochondria and more vacuolar and broken cell membrane were found in TEM after the treatment of hypocrellin B and ultrasound. Our findings demonstrated that the presence of hypocrellin B significantly enhanced the cytotoxicity of ultrasound radiation in CNE2 cells, suggesting that hypocrellin B is a novel sonosensitizer and hypocrellin B-mediated sonodynamic therapy is a potential therapeutic modality in the management of malignant tumors. PMID- 20018429 TI - Nonlinear emission from individual bound microbubbles at high frequencies. AB - Targeted microbubbles detected with high-frequency ultrasound can establish the molecular expression of blood vessels with submillimeter resolution. To improve microbubble-specific imaging at high frequencies, the subharmonic and second harmonic signal from individual microbubbles were measured as a function of size and pressure. Single phospholipid-shell microbubbles (1.1 to 5.0 microm in diameter) bound to gelatin, co-aligned with an optical microscope and transducer, were insonated with 30 MHz Gaussian-enveloped pulses at pressures from 20 kPa to 1 MPa with -6 dB one-way bandwidths of 11%, 20% and 45%. A subharmonic signal (15 MHz) was detected above a pressure threshold of 110 kPa--independent of bandwidth. The signal peaked for microbubbles 1.60 microm in diameter subject to 20% and 11% bandwidth pulses, and 1.80 microm for 45% bandwidth pulses, for pressures up to 400 kPa, agreeing with the notion that microbubbles insonated at twice their resonant frequency preferentially emit a subharmonic component. For pressures between 400 kPa and 1 MPa, a broader range of microbubbles emitted a subharmonic signal, and microbubbles below 1.70 mum in diameter were disrupted. The second harmonic signal measured, within the limited experimental conditions, was consistent with nonlinear propagation. Further, the results shed light on the effect of the shell on the phase of the subharmonic signal with respect to the fundamental signal. PMID- 20018430 TI - Percentage white: a new feature for ultrasound classification of plaque echogenicity in carotid artery atherosclerosis. AB - Atherosclerotic stenotic and nonstenotic plaques of the carotid artery with low echogenicity have been shown to be associated with cardiovascular disease. The aim was to develop a new method for semiautomated ultrasound image analysis to classify nonstenotic carotid plaques, evaluate cases with multiple plaques and examine the association between a new image analysis feature of echogenicity and predictors of cardiovascular disease. The new image analysis feature, percentage white (PW), represents the fraction of bright structures inside a plaque and is integrated in an objective semiautomated method to evaluate echogenicity (SAMEE) in carotid plaques. PW was constructed to take into account overall echogenicity of the image as well as noise surrounding the plaque. Consecutive ultrasound examinations of carotid plaques from a population-based screening of 64-year-old women with varying risk for cardiovascular disease were selected for the present project; 92 far-wall and 47 near-wall plaques were used as a training dataset to develop the SAMEE algorithm with visual classification according to Gray-Weale as reference; 273 plaques were used to validate the method. All plaques were included in an analysis relating predictors of cardiovascular to average PW in all plaques, PW in the biggest plaque and to the plaque with lowest PW in each subject, respectively. In the training dataset the intermethodological variability between SAMEE and visual classification showed a kappa of 0.78 and a sensitivity and specificity of 96% and 81%, respectively. In the validation set, SAMEE and visual classification showed a kappa of 0.77, a sensitivity of 96% and a specificity of 80%. The reproducibility of PW was high, evidenced by r = 0.96 and CV = 9.85% at repeated examinations. Average PW values were associated with several predictors of cardiovascular risk: lipoprotein (a), HbA1c, blood glucose, apolipoproteinB/apolipoproteinA-I; and associated negatively with the levels of adiponectin and apolipoprotein A-I. In conclusion, PW integrated within a SAMEE is a new feature for assessment of echogenicity in carotid plaques and shows excellent reproducibility and agreement with visual assessment. PMID- 20018432 TI - Acoustic radiation force impulse elastography for the evaluation of focal solid hepatic lesions: preliminary findings. AB - This study was designed to investigate the potential usefulness of acoustic radiation force impulse (ARFI) elastography to evaluate focal solid hepatic lesions. In total, 51 patients with 60 focal hepatic lesions, which included 17 hemangiomas, 25 hepatocellular carcinomas (HCCs), 15 metastases and three cholangiocarcinomas, underwent ARFI elastography. The lesions were classified into three groups: Group I consisted of metastatic liver tumors and cholangiocarcinomas, group II consisted of HCCs and group III consisted of hemangiomas. The stiffness and conspicuity of the tumors as depicted on ARFI elastography and the echogenicity and conspicuity of the tumors on corresponding B-mode images were analyzed. Shear wave velocity was obtained to quantify stiffness for 36 focal hepatic lesions: 11 hemangiomas, 17 HCCs and eight other malignant lesions. On ARFI elastography images, group I tumors (n=18) appeared stiffer than the background liver for 13 lesions (72%), softer for two lesions and had identical stiffness in three lesions compared with the background liver. For group II tumors (n=25), 13 lesions (52%) appeared stiffer than the liver, six lesions appeared softer than the liver and the remaining six lesions showed the same stiffness as the liver. For group III tumors (n=17), six lesions (35%) appeared stiffer than the liver, seven lesions appeared softer and the remaining four lesions showed the same stiffness as the liver. There were no statistical differences among the three groups in terms of tumor stiffness as seen on ARFI elastography images (p>0.05). Of the 60 lesions, 41 (68%) displayed a clearer or equivalent margin on the ARFI elastography compared with that seen on B-mode images. The shear wave velocities were: Group I, 2.18+/-0.96 m/s (mean value+/ SD); group II, 2.45+/-0.81m/s; group III, 1.51+/-0.71 m/s (p=0.012). With a cut off value of 2m/s for the shear wave velocity, the positive predictive value and specificity for malignancy were 89% and 81%, respectively. Images obtained with ARFI elastography provided additional qualitative information regarding the stiffness and tumor margin of liver tumors. By measuring shear wave velocity, quantification of stiffness was made possible and showed the potential to differentiate malignant hepatic tumors from hepatic hemangiomas. PMID- 20018433 TI - Shock-induced heating and millisecond boiling in gels and tissue due to high intensity focused ultrasound. AB - Nonlinear propagation causes high-intensity ultrasound waves to distort and generate higher harmonics, which are more readily absorbed and converted to heat than the fundamental frequency. Although such nonlinear effects have been investigated previously and found to not significantly alter high-intensity focused ultrasound (HIFU) treatments, two results reported here change this paradigm. One is that at clinically relevant intensity levels, HIFU waves not only become distorted but form shock waves in tissue. The other is that the generated shock waves heat the tissue to boiling in much less time than predicted for undistorted or weakly distorted waves. In this study, a 2-MHz HIFU source operating at peak intensities up to 25,000 W/cm(2) was used to heat transparent tissue-mimicking phantoms and ex vivo bovine liver samples. Initiation of boiling was detected using high-speed photography, a 20-MHz passive cavitation detector and fluctuation of the drive voltage at the HIFU source. The time to boil obtained experimentally was used to quantify heating rates and was compared with calculations using weak shock theory and the shock amplitudes obtained from nonlinear modeling and measurements with a fiber optic hydrophone. As observed experimentally and predicted by calculations, shocked focal waveforms produced boiling in as little as 3 ms and the time to initiate boiling was sensitive to small changes in HIFU output. Nonlinear heating as a result of shock waves is therefore important to HIFU, and clinicians should be aware of the potential for very rapid boiling because it alters treatments. PMID- 20018434 TI - Critical appraisal of targeted ultrasound contrast agents for molecular imaging in large arteries. AB - Molecular imaging may provide new insights into the early detection and development of atherosclerosis before first symptoms occur. One of the techniques in use employs noninvasive ultrasound. In the past decade, experimental and clinical validation studies showed that for the microcirculation targeted ultrasound contrast agents, such as echogenic liposomes, microbubbles and perfluorocarbon emulsions, do improve visualization of specific structures. For large arteries, however, successful application is less obvious. In this review, we will address the challenges for molecular imaging of large arteries. We will discuss the problems encountered in the use of targeted ultrasound contrast agents presently available, mainly based on data obtained in flow chambers and animal studies because clinical studies are lacking. We conclude that molecular imaging of activated endothelium in large- and middle-sized arteries by site specific accumulation of contrast material is still difficult to achieve due to wall shear stress conditions in these vessels. PMID- 20018435 TI - Opening of the blood-brain barrier by low-frequency (28-kHz) ultrasound: a novel pinhole-assisted mechanical scanning device. AB - Disruption of the blood-brain barrier (BBB) may be transiently achieved via high frequency focused spherical ultrasound in the presence of microbubbles. In this experimental animal study, we sought to determine whether focal reversible opening of the BBB may be achieved using low-frequency (i.e., 20-30 kHz) planar ultrasonic waves. In the presence of microbubbles, we were able to obtain BBB opening using non-focused ultrasound irradiation with a frequency as low as 28 kHz. We also achieved a tight regulation of the ultrasound patterns by using a mechanical scanning device equipped with a pinhole. Histologic examination of the brains supported the feasibility of our system. The areas of BBB disruption obtained with this method were large enough to cover a typical circumscribed cerebral tumor mass. The inherent advantages of our BBB opening method include an improved portability, the possibility to obtain fairly wide areas of BBB opening and a low incidence of hemorrhagic complications. In addition, our system has the potential to reduce the need for image guidance for treating superficial brain lesions. PMID- 20018436 TI - Ultrasonic Nakagami imaging: a strategy to visualize the scatterer properties of benign and malignant breast tumors. AB - Previous studies have demonstrated the usefulness of the Nakagami parameter in characterizing breast tumors by ultrasound. However, physicians or radiologists may need imaging tools in a clinical setting to visually identify the properties of breast tumors. This study proposed the ultrasonic Nakagami image to visualize the scatterer properties of breast tumors and then explored its clinical performance in classifying benign and malignant tumors. Raw data of ultrasonic backscattered signals were collected from 100 patients (50 benign and 50 malignant cases) using a commercial ultrasound scanner with a 7.5 MHz linear array transducer. The backscattered signals were used to form the B-scan and the Nakagami images of breast tumors. For each tumor, the average Nakagami parameter was calculated from the pixel values in the region-of-interest in the Nakagami image. The receiver operating characteristic (ROC) curve was used to evaluate the clinical performance of the Nakagami image. The results showed that the Nakagami image shadings in benign tumors were different from those in malignant cases. The average Nakagami parameters for benign and malignant tumors were 0.69 +/- 0.12 and 0.55 +/- 0.12, respectively. This means that the backscattered signals received from malignant tumors tend to be more pre-Rayleigh distributed than those from benign tumors, corresponding to a more complex scatterer arrangement or composition. The ROC analysis showed that the area under the ROC curve was 0.81 +/- 0.04 and the diagnostic accuracy was 82%, sensitivity was 92% and specificity was 72%. The results showed that the Nakagami image is useful to distinguishing between benign and malignant breast tumors. PMID- 20018437 TI - Head-to-head comparison of the three most commonly used preoperative models for prediction of biochemical recurrence after radical prostatectomy. AB - BACKGROUND: Several models can predict the rate of biochemical recurrence (BCR) after radical prostatectomy (RP). OBJECTIVE: We tested the three most commonly used models-the D'Amico risk stratification scheme, the Cancer of the Prostate Risk Assessment (CAPRA) score, and the Stephenson nomogram-in a European cohort of RP patients. DESIGN, SETTING, AND PARTICIPANTS: We relied on preoperative characteristics and prostate-specific antigen follow-up data of 1976 patients, as required by the three tested models. All patients were treated with an open RP between 1992 and 2006. MEASUREMENTS: Analyses included tests of accuracy (Harrell's concordance index) and calibration between predicted and observed BCR rates at 3 yr and 5 yr after RP. Additionally, we relied on decision curve analyses to compare the three models directly in a head-to-head fashion. RESULTS AND LIMITATIONS: The median follow-up of censored patients was 32 mo. BCR-free rates at 3 yr and 5 yr after RP were 80.2% and 72.6%, respectively. The concordance index for 3-yr BCR predictions was 70.4%, 74.3%, and 75.2% for the D'Amico, CAPRA, and Stephenson models, respectively, versus 67.4%, 72.9%, and 73.5% for 5-yr BCR predictions. Calibration results supported the use of either the CAPRA or Stephenson models. Decision curve analyses indicated a small benefit for the CAPRA score relative to the Stephenson nomogram. Our findings apply to German patients treated with RP at a high-volume tertiary care centre. Consequently, the rank order reported in this paper may not be the same in North American or other European cohorts. CONCLUSIONS: Different methods yield different results, and it may be difficult to reconcile concordance index, calibration, and decision curve analysis findings. Our data suggest that the CAPRA score outperforms the other models when decision curve analysis and calibration were used as benchmarks. Conversely, the Stephenson nomogram outperformed the other models when concordance index was used as a metric. PMID- 20018438 TI - Should bladder cuff excision remain the standard of care at nephroureterectomy in patients with urothelial carcinoma of the renal pelvis? A population-based study. AB - BACKGROUND: A large, multi-institutional, tertiary care center study suggested no benefit from bladder cuff excision (BCE) at nephroureterectomy in patients with upper tract urothelial carcinoma (UC). OBJECTIVE: We tested and quantified the prognostic impact of BCE at nephroureterectomy on cancer-specific mortality (CSM) in a large population-based cohort of patients with UC of the renal pelvis. DESIGN, SETTING, AND PARTICIPANTS: A cohort of 4210 patients with UC of the renal pelvis were treated with nephroureterectomy with (NUC) or without (NU) a BCE between 1988 and 2006 within 17 Surveillance, Epidemiology, and End Results registries. MEASUREMENTS: Cumulative incidence plots and competing risks regression models compared CSM after either NUC or NU. Covariates consisted of pathologic T and N stages, grade, age, year of surgery, gender, and race. RESULTS AND LIMITATIONS: Respectively, 2492 (59.2%) and 1718 (40.8%) patients underwent a nephroureterectomy with or without BCE. In univariable and multivariable analyses, BCE omission increased CSM rates in patients with pT3N0/x, pT4N0/x, and pT(any)N1-3 UC of the renal pelvis. For example, in patients with pT3N0/x disease, holding all other variables constant, BCE omission increased CSM in a 1.25-fold fashion (p=0.04). Similarly, in patients with pT4N0/x disease, BCE omission resulted in a 1.45-fold increase (p=0.02). The main limitation of our study is the lack of data on disease recurrence. CONCLUSIONS: Nephroureterectomy with BCE remains the standard of care in the treatment of UC of the renal pelvis and should invariably be performed in patients with locally advanced disease. Conversely, patients with pT1 and pT2 disease could be considered for NU without compromising CSM. However, recurrence data are needed to fully confirm the validity of this option. PMID- 20018439 TI - Management of urethral stent failure for recurrent anterior urethral strictures. AB - BACKGROUND: Urethral stent placement for recurrent anterior urethral strictures may cause restenosis and complications. OBJECTIVE: To describe our experience with patients who had restenoses and complications following urethral stent placement for the treatment of recurrent anterior urethral strictures. DESIGN, SETTING, AND PARTICIPANTS: We evaluated retrospectively the records of 13 men with anterior urethral stricture who experienced restenosis and complications after stent insertion. We recorded stent position, prestent and poststent urethral procedures, restenosis location, stent-related complications, and management of stent failures. SURGICAL PROCEDURE: The stent was removed en bloc with the whole strictured urethral segment or wire by wire after a ventral or a double-ventral plus dorsal-sagittal urethrotomy and stent section. MEASUREMENTS: Successful outcome was defined as standard voiding, without need of any postoperative procedure, and full recovery from complications. RESULTS AND LIMITATIONS: Four patients did not undergo surgery and the stent was left in situ. Of these patients, two required permanent suprapubic cystostomy. Nine patients underwent challenging surgical stent removal and salvage urethrostomy: After the first stage, three patients are waiting for further reconstructive steps, five elected the urethrostomy as a permanent diversion, and one completed the staged reconstruction using a buccal mucosa graft at the second stage. After surgery, seven of the nine patients (77.8%) were free of strictures and stent related complications, while a restenosis occurred in two of the nine (22.2%) cases. CONCLUSIONS: The management of urethral stent failure represents a therapeutic challenge. The stent risks converting a simple stenosis into a complex stenosis requiring a staged urethroplasty, a definitive urethrostomy, or a permanent suprapubic diversion. PMID- 20018440 TI - Could Helicobacter pylori play an important role in axonal type of Guillain-Barre syndrome pathogenesis? AB - In this case-control study, ELISA and Western blot with whole bacterial protein lysate were performed on serum and cerebrospinal fluid (CSF) of 15 controls and 15 patients. According to Griffin subtypes, 10 of our patients were in acute inflammatory demyelinating polyradiculoneuropathy (AIDP) group, 3 in acute motor axonal neuropathy (AMAN) group, and 2 in acute motor sensory axonal neuropathy (AMSAN) subtype. 86.6% of patients were positive for Helicobacter pylori (H. pylori) IgG. Serum anti-H. pylori IgG of patients and controls were significantly different. CSF anti-H. pylori IgG was significantly higher in patients than controls. In patients, the titer of anti-H. pylori IgG in serum was significantly higher than CSF, which may indicate extra-neural antibody synthesis. CSF IgG titer was higher in patients having axonal pattern. Western blot was positive in CSF of 13 patients and negative in all controls. There was a correlation between the number of antibody types against H. pylori particles and the titer of anti-H. pylori IgG in CSF and serum. Also, antibody against cytotoxin associated protein (CagA) was associated with primary axonal pattern. The association between the presence of anti-CagA and primary axonal pattern, is in favor of the relation between axonal neuropathy and H. pylori infection. PMID- 20018441 TI - Isolated degeneration of the posterior column as a distinct entity--a clinical and electrophysiologic follow-up study. AB - OBJECTIVE: The aim of the study was to better describe the long term clinical course and electrophysiologic and radiologic findings in isolated degeneration of the posterior column. METHODS: Four patients with the presenting symptoms of a progressive tabetic ataxia were followed up clinically and electrophysiologically over up to 15 years between 1997 and 2008. They received standardized neurological examinations, electrophysiologic testing with SEP, MEP, NCV, EMG, autonomic testing and cardiac evaluation, head and spine MRI, laboratory evaluation including CSF analysis. RESULTS: Progressive gait ataxia due to pallhypasthesia and loss of position sense with areflexia remained the only symptoms. Pes cavus deformity was a notable clinical feature in all cases. There was no involvement of other systems and all patients remained fully ambulatory. There was no cardiac involvement. Electrophysiology was characterized by absent cortical tibial SEP with normal lumbar complexes and normal nerve conduction studies and transcortical magnetic stimulation as well as sympathetic skin response. MRI of the cord was normal. Laboratory analysis and CSF were unrevealing. CONCLUSION: Isolated degeneration of the posterior column is a rare condition with a clinically benign course without progression involving other systems and characteristic electrophysiologic findings (isolated loss of cortical tibial-SEP with normal lumbar leads). Pes cavus deformity seems to be an unusual but typical clinical feature. The etiology is most likely a sporadic degenerative disease of the cord. PMID- 20018442 TI - Hypoxia suppresses chemotherapeutic drug-induced p53 Serine 46 phosphorylation by triggering HIPK2 degradation. AB - The molecular mechanisms by which hypoxic tumor cells escape radio- and chemotherapy are largely unclear. Homeodomain-interacting protein kinase 2 (HIPK2) drives the apoptotic program in response to DNA-damaging chemotherapeutic drug treatment by phosphorylating the tumor suppressor protein p53 at Ser46. HIPK2 is kept inactive in unstressed cells through ubiquitination and degradation facilitated by the ubiquitin ligases WSB1 and Siah1. Here, we demonstrate that HIPK2 is degraded during hypoxia in a proteasome-dependent and partially Siah1 dependent fashion. Concordantly, hypoxic tumor cells show an impaired p53 Ser46 phosphorylation in response to treatment with the chemotherapeutic Adriamycin. Remarkably, proteasome-inhibition rescues HIPK2 expression in hypoxic hepatoma cells and restores p53 Ser46 phosphorylation and caspase activity after Adriamycin treatment. Our findings suggest a molecular mechanism by which hypoxic cancer cells can escape chemotherapeutic drug treatment and suggest proteasome inhibition as a promising approach to sensitise hypoxic cancer cells to therapy. PMID- 20018443 TI - Spatial characterization of dissolved trace elements and heavy metals in the upper Han River (China) using multivariate statistical techniques. AB - A data matrix (4032 observations), obtained during a 2-year monitoring period (2005-2006) from 42 sites in the upper Han River is subjected to various multivariate statistical techniques including cluster analysis, principal component analysis (PCA), factor analysis (FA), correlation analysis and analysis of variance to determine the spatial characterization of dissolved trace elements and heavy metals. Our results indicate that waters in the upper Han River are primarily polluted by Al, As, Cd, Pb, Sb and Se, and the potential pollutants include Ba, Cr, Hg, Mn and Ni. Spatial distribution of trace metals indicates the polluted sections mainly concentrate in the Danjiang, Danjiangkou Reservoir catchment and Hanzhong Plain, and the most contaminated river is in the Hanzhong Plain. Q-model clustering depends on geographical location of sampling sites and groups the 42 sampling sites into four clusters, i.e., Danjiang, Danjiangkou Reservoir region (lower catchment), upper catchment and one river in headwaters pertaining to water quality. The headwaters, Danjiang and lower catchment, and upper catchment correspond to very high polluted, moderate polluted and relatively low polluted regions, respectively. Additionally, PCA/FA and correlation analysis demonstrates that Al, Cd, Mn, Ni, Fe, Si and Sr are controlled by natural sources, whereas the other metals appear to be primarily controlled by anthropogenic origins though geogenic source contributing to them. PMID- 20018444 TI - Modeling solid thermal explosion containment on reactor HNIW and HMX. AB - 2,4,6,8,10,12-Hexanitro-2,4,6,8,10,12-hexaaza-isowurtzitane (HNIW), also known as CL-20 and octahydro-1,3,5,7-tetranitro-1,3,5,7-tetrazocine (HMX), are highly energetic materials which have been popular in national defense industries for years. This study established the models of thermal decomposition and thermal explosion hazard for HNIW and HMX. Differential scanning calorimetry (DSC) data were used for parameters determination of the thermokinetic models, and then these models were employed for simulation of thermal explosion in a 437L barrel reactor and a 24 kg cubic box package. Experimental results indicating the best storage conditions to avoid any violent runaway reaction of HNIW and HMX were also discovered. This study also developed an efficient procedure regarding creation of thermokinetics and assessment of thermal hazards of HNIW and HMX that could be applied to ensure safe storage conditions. PMID- 20018445 TI - Investigation of dye functional group on the photocatalytic degradation of dyes by nano-TiO2. AB - The photocatalytic degradation of five anionic, eight cationic and three solvent dyes using combustion-synthesized nano-TiO(2) (CS TiO(2)) and commercial Degussa P-25 TiO(2) (DP-25) were evaluated to determine the effect of the functional group in the dye. The degradation of the dyes was quantified using the initial rate of decolorization and mineralization. The decolorization of the anionic dyes with CS TiO(2) followed the order: indigo carmine > eosin Y > amido black 10B > alizarin cyanine green > orange G. The decolorization of the cationic dyes with DP-25 followed the order: malachite green > pyronin Y > rhodamine 6G > azure B > nile blue sulfate > auramine O approximately = acriflavine approximately = safranin O. CS TiO(2) showed higher rates of decolorization and mineralization for all the anionic dyes compared to DP-25, while DP-25 was better in terms of decolorization for most of the cationic dyes. The solvent dyes exhibited adsorption dependent decolorization. The order of decolorization and mineralization of the anionic and cationic dyes (a) with CS TiO(2) and DP-25 was different and correlated with the surface properties of these catalysts (b) were rationalized with the molecular structure of the dye and the degradation pathway of the dye. PMID- 20018446 TI - Isotherm and kinetics study for acrylic acid removal using powdered activated carbon. AB - The potential of powdered activated carbon (PAC) for the adsorption of acrylic acid (AA) from aqueous solution was studied at the initial concentration (C(0)) in the range of 50-500 mg/l over the temperature range of 303-348 K. The equilibrium adsorption studies were carried out to evaluate the effect of adsorbent dosage and contact time, change in pH by adding adsorbents and the initial concentration. Langmuir, Freundlich and Redlich-Peterson (R-P) equilibrium isotherm models were tested to represent the data. Error functions were used to test their validity to fit of the adsorption data with the isotherm and kinetic models. The Freundlich isotherm equation is found to best represent the equilibrium separation data in the temperature range of 303-348 K. The maximum adsorption capacity of AA onto PAC was obtained as q(m)=36.23 mg/g with an optimum PAC dosage w=20 g/l at 303 K for C(0)=100 mg/l. The pseudo-second order kinetics is found to represent the experimental AA-PAC data. The negative value of DeltaG(ad)(o) (-16.60 to -18.18 kJ/mol K) indicate the feasibility and spontaneity of the adsorption process. PMID- 20018447 TI - Selection of best impregnated palm shell activated carbon (PSAC) for simultaneous removal of SO2 and NOx. AB - This work examines the impregnated carbon-based sorbents for simultaneous removal of SO(2) and NOx from simulated flue gas. The carbon-based sorbents were prepared using palm shell activated carbon (PSAC) impregnated with several metal oxides (Ni, V, Fe and Ce). The removal of SO(2) and NOx from the simulated flue gas was investigated in a fixed-bed reactor. The results showed that PSAC impregnated with CeO(2) (PSAC-Ce) reported the highest sorption capacity among other impregnated metal oxides for the simultaneous removal of SO(2) and NOx. PSAC-Ce showed the longest breakthrough time of 165 and 115 min for SO(2) and NOx, respectively. The properties of the pure and impregnated PSAC were analyzed by BET, FTIR and XRF. The physical-chemical features of the PSAC-Ce sorbent indicated a catalytic activity in both the sorption of SO(2) and NOx. The formation of both sulfate (SO(4)(2-)) and nitrate (NO(3-)) species on spent PSAC Ce further prove the catalytic role played by CeO(2). PMID- 20018448 TI - Nickel recovery from spent Raneynickel catalyst through dilute sulfuric acid leaching and soda ash precipitation. AB - Pharmaceutical industry makes extensive use of Raneynickel catalyst for various organic drug intermediates/end products. Spent catalysts contain environmentally critical and economically valuable metals. In the present study, a simple hydrometallurgical process using dilute sulfuric acid leaching was described for the recovery of nickel from spent Raneynickel catalyst. Recovery of nickel varied with acid concentration and time, whereas temperature had negligible effect. Increase of S/L ratio to 30% (w/v) showed marginal effect on nickel (90%) recovery, whereas Al recovery decreased drastically to approximately 20%. Under the optimum conditions of leaching viz: 12 vol.% H(2)SO(4), 30 degrees C, 20% solid to liquid (S/L) ratio and 120 min reaction time, it was possible to recover 98.6% Ni along with 39.2% Al. Leach liquor [pH 0.7] containing 85.0 g/L Ni and 3.25 g/L Al was adjusted to pH 5.4 with 30 wt.% alkali for quantitative aluminum removal. Nickel loss was about 2% during this Al removal step. Nickel from the purified leach liquor was recovered as nickel carbonate by adding required amount of Na(2)CO(3). The purity of NiCO(3) product was found to be 100% with a Ni content of 48.6%. Na(2)SO(4) was recovered as a by-product with a purity of 99%. Complete process is presented. PMID- 20018449 TI - Co-treatment of olive-mill and urban wastewaters by experimental stabilization ponds. AB - Olive oil mill wastewater (OMW) constitutes a source of environmental problems in Morocco due to its significantly high organic load, its phytotoxic properties and its relatively low biodegradability. An effective option for its disposal is its agricultural use after co-treatment with urban wastewater (UWW). The main objective of this investigation was to evaluate the potential of this co treatment, using experimental waste stabilization ponds, in removing OMW phytotoxicity. We examined the influence of the organic load, at the entry of the treatment system, on the evolution of some physicochemical (chemical oxygen demand and polyphenols) and microbiological (fecal coliforms and fecal streptococci) parameters. The results showed a removal of the organic, phenolic and microbial load throughout the treatment which differed from one system to another according to the OMW load applied to each system. The results concerning the germination assays of Zea mays and Solanum lycopersicum suggested that the co treatment of OMW with UWW would decrease the phytotoxicity of this waste. PMID- 20018450 TI - Radiation dose estimation and mass attenuation coefficients of cement samples used in Turkey. AB - Different cement samples commonly used in building construction in Turkey have been analyzed for natural radioactivity using gamma-ray spectrometry. The mean activity concentrations observed in the cement samples were 52, 40 and 324 Bq kg( 1) for (226)Ra, (232)Th and (40)K, respectively. The measured activity concentrations for these radionuclides were compared with the reported data of other countries and world average limits. The radiological hazard parameters such as radium equivalent activities (Ra(eq)), gamma index (I(gamma)) and alpha index (I(alpha)) indices as well as terrestrial absorbed dose and annual effective dose rate were calculated and compared with the international data. The Ra(eq) values of cement are lower than the limit of 370 Bq kg(-1), equivalent to a gamma dose of 1.5 mSv y(-1). Moreover, the mass attenuation coefficients were determined experimentally and calculated theoretically using XCOM in some cement samples. Also, chemical compositions analyses of the cement samples were investigated. PMID- 20018451 TI - Burn resuscitation: the results of the ISBI/ABA survey. AB - INTRODUCTION: There are valid concerns that burn shock resuscitation is inadequate; a tendency to over-resuscitate the patient seems to exist which may increase complications such as compartment syndrome. The purpose of this study was to survey members of the ISBI and ABA to determine current practices of burn resuscitation. METHODS: A survey asking for practices of burn shock resuscitation was provided to all participants of a recent ABA meeting. Around the same time, the survey was sent to all members of the ISBI through the internet. The results of the 101 respondents (ABA--59, ISBI--42, approximately a 15% response rate) are described. RESULTS: Surveys were returned from all the continents except Africa. Respondents included directors (48%), staff physicians (19%), nurses (23%) and others. Most programs admitted adults (87%) and children (75%) with a mean of 289 admissions per year. The cut off to initiate resuscitation was 15% TBSA and most preferred peripheral IVs (70%) and central lines (47.5%). The Parkland formula was preferred (69.3%) while others were used: Brooke--6.9%, Galveston--8.9%, Warden--5.9%, and colloid 11.9%. Lactated Ringer's (LR) was the preferred solution (91.9%), followed by normal saline--5%, hypertonic saline--4%, albumin- 20.8%, FFP--13.9%, and LR/NaHCO(3)--12.9%. Approximately half (49.5%) added colloid before 24h. Urine output is the major indicator of success (94.9%) while 22.7% use other monitors. Most (88.8%) feel their protocols work well, with 69.8% feel that it provides the right amount of fluid (24%--too much, 7%--too little). Despite this feeling, they still feel that they give more fluid than the formula in 55.1%, less than formula in 12.4% and the right amount in 32.6%. Approximately 1/3 use an oral resuscitation formula and 81.8% feel that an oral formula works for burns<15% TBSA. CONCLUSION: Large variations exist in resuscitation protocols but the Parkland formula using LR is still the dominant method. Most feel that their resuscitation protocol works well. PMID- 20018452 TI - Treatment of the pregnant mother with cancer: a systematic review on the use of cytotoxic, endocrine, targeted agents and immunotherapy during pregnancy. Part II: Hematological tumors. AB - Managing pregnant patients with hematological tumors pose even more conflicts compared to solid tumors. Unlike the majority of solid tumors, hematological malignancies are potentially curable; hence it is important to deliver the best treatment options available, which sometimes could be too aggressive to deliver during pregnancy. In part II, we report the results of women with hematological malignancies treated with systemic therapies during the course of pregnancy. Lymphoma, acute leukemia and chronic myeloid leukemia were the most commonly treated. We discuss the safety of the different regimens reported and propose alternatives to standardized approaches in case they pose significant risk to the pregnancy and/or the fetus. PMID- 20018453 TI - After science: has the tradition been broken? AB - The majority of professional scientists make use of the artefacts of science but lack understanding of what these mean; raising the question: has the tradition of science been broken? Explicit knowledge is only a selective summary but practical capability derives from implicit, traditional or 'tacit' knowledge that is handed on between- and across-generations by slow, assimilative processes requiring extended human contact through a wide range of situations. This was achieved mainly by prolonged apprenticeship to a Master. Such methods recognize the gulf between being able to do something and knowing how you have done it; and the further gap between knowing how you have done something and being able to teach it by explicit instructions. Yet the 'Master-apprentice' model of education has been almost discarded from science over recent decades and replaced with bureaucratic regulation. The main reason is probably that scientific manpower has expanded so rapidly and over such a long period as to overwhelm the slow, sure and thorough traditional methods. In their innocence of scientific culture, the younger generation of scientists are like children who have been raised by wolves; they do not talk science but spout bureaucratic procedures. It has now become accepted among the mass of professional 'scientists' that the decisions which matter most in science are those imposed upon science by outside forces: for example by employers, funders, publishers, regulators, and the law courts. It is these bureaucratic mechanisms that now constitute the 'bottom line' for scientific practice. Most of modern science is therefore apparently in the post holocaust situation described in A canticle for Liebowitz and After Virtue, but the catastrophe was bureaucratic, rather than violent. So, the tradition has indeed been broken. However, for as long as the fact is known that the tradition has been broken, and living representatives of the tradition are still alive and active, there still exists a remote possibility that the tradition could be revived. PMID- 20018454 TI - Historical "evidence" that electrification caused the 20th century epidemic of diseases of civilization and the ecological fallacy. PMID- 20018456 TI - Why high frequency of distraction improved the bone formation in distraction osteogenesis? AB - Distraction osteogenesis, currently a standard method of bone lengthening, is based upon the "tension-stress principle", as proposed by G.A. Ilizarov. Mechanical stimulation by distraction induces biological responses of skeletal regeneration that is accomplished by a cascade of biologic processes including differentiation of pluripotential tissue, angiogenesis, mineralization, and remodeling. The exact mechanism by which strain stimulates bone formation remains unclear. Distraction rate and rhythm must have great influence on the quality of the newly formed bone generated by mechanical traction. The preliminary results demonstrated that for a given rate higher frequency of distraction improved the bone formation, but the mechanism remains unclear. In this article we present a hypothesis that the reason why higher frequency of distraction improved the bone formation for a given rate is that higher frequency of distraction provides smaller microtrauma to tissues within the gap and longer existence time of the microenvironment stimulating tissues within the gap than low frequency distraction. This hypothesis, if proven to be valid, will not only represent a breakthrough in research of mechanism of distraction osteogenesis, but also will open a new door to the bone regeneration. PMID- 20018455 TI - Serotonin, pregnancy and increased autism prevalence: is there a link? AB - The prevalence of autism, a neurodevelopmental condition resulting from genetic and environmental causes, has increased dramatically during the last decade. Among the potential environmental factors, hyperserotonemia during pregnancy and its effect on brain development could be playing a role in this prevalence raise. In the rodent model developed by Whitaker-Azmitia and colleagues, hyperserotonemia during fetal development results in a dysfunction of the hypothalamo-pituitary axis, affecting the amygdala as well as pro-social hormone oxytocin regulation. Dysfunction of the amygdala and abnormal oxytocin levels may underlie many clinical features of ASD. Selective serotonin reuptake inhibitors (SSRI) are the most widely used class of antidepressants drugs, and they are not contraindicated during pregnancy. In this paper, we hypothesize that increased serotonemia during pregnancy, including due to SSRI intake, could be one of the causes of the raising prevalence in autism. If our hypothesis is confirmed, it will not only shed light on one of the possible reason for autism prevalence, but also offer new preventive and treatment options. PMID- 20018457 TI - Computed tomography perfusion assessment of radiation therapy effects on spinal cord hemodynamics. AB - PURPOSE: We used computed tomography (CT) perfusion to evaluate the acute and late effect of radiation therapy (RT) on spinal cord (SC) hemodynamics in patients without symptoms of myelopathy. We hypothesized that SC perfusion could be acutely altered during RT. METHODS AND MATERIALS: We analyzed neck CT perfusion studies of 36 head-and-neck cancer patients (N1), 16 of whom had previously undergone RT. In a separate group of 6 patients (N2), CT perfusion studies were obtained before RT, after 40 Gy, and after treatment completion. RESULTS: In the N1 group, SC blood flow (BF), blood volume (BV), mean transit time (MTT), and capillary permeability (CP) maps were not significantly different between RT-treated and RT-naive patients. In the N2 group, BF and CP were significantly increased during treatment compared with the baseline and post-RT studies. CONCLUSIONS: Radiation therapy of the head and neck may cause transient perturbations of SC perfusion that seem to reverse after treatment. There are no definite chronic effects of RT on SC perfusion observeable at the typical doses administered during treatment of head and neck malignancies. PMID- 20018458 TI - [Miniarc : prospective study and follow up at one year about 72 patients]. AB - OBJECTIVE: To evaluate the results at 1 year of a mini invasive sub urethral sling, not requiring no more a single incision. PATIENTS AND METHODS: Prospective study of 72 patients with isolated stress urinary incontinence. The procedures were performed on an outpatient under local anesthesia. Pain was assessed using the visual analogue scale. All patients were checked at 2 months and 1 year of intervention. RESULTS: Stress urinary incontinence for 58 patients, 14 for mixed incontinence and sphincter deficiency for five. As per procedure, the average pain was 3.7/10, at the end of the intervention, 0.5/10. Peroperative complications were: one episode involving bleeding over 100ml, one vasovagal episode, and one procedure failure. At 2 months follow-up, the authors recorded: de novo urge incontinence (n=2), de novo dysuria (n=1), sling exposure (n=1), urinary infection (n=1) and noticeable side ties (n=4). Control at 1 year, another patient filed an eviction of the anchor of the tape with Stall thereof. Two patients reported having moderate pain. The objective results at 1 year of 68 patients were 69.1% cured, 20.6% improved and 10.3% failed. Three patients were re-operated and two of them are cured. CONCLUSION: The results of this simplified tape are worse than those of classic sub-urethral tape; however, no comparative study has been completed and the specific indications of these techniques are not specified. PMID- 20018461 TI - Experimental nebulization of American bison (Bison bison) with low doses of ovine herpesvirus 2 from sheep nasal secretions. AB - Malignant catarrhal fever (MCF), caused by ovine herpesvirus 2 (OvHV-2), is an important cause of mortality in ranched American bison and domestic cattle in North America. Previous studies showed that bison can be infected by intranasal nebulization with sheep nasal secretions containing OvHV-2 and provided preliminary information on viral doses required for infection and disease progression. The goals of this study were to establish optimal minimal infectious and minimal lethal doses of OvHV-2 by the intranasal route in bison, evaluate the influence of dose on incubation period and other clinical parameters and determine if bison seropositive for antibody against MCF-group viruses are resistant to developing MCF after intranasal challenge. In this study, the minimal infectious dose and minimal lethal dose overlap, suggesting that experimental production of subclinically infected bison is impractical. Dose is inversely related to both incubation period and the period between nebulization and first detection of >1000 OvHV-2 DNA copies/500 ng total DNA in peripheral blood leukocytes. Interestingly, all of the bison seropositive for anti-MCF-group viral antibody prior to inoculation died of MCF after nebulization. We conclude that previous exposure to an MCF-group virus does not necessarily provide resistance to OvHV-2-induced MCF in bison. PMID- 20018462 TI - Bartonellosis. AB - Bartonellosis is a constellation of clinical conditions affecting human beings and a variety of animals. Many Bartonella infections are zoonotic, with some of the most commonly reported zoonotic manifestations of infection including cat scratch disease, bacillary angiomatosis, endocarditis, and neuroretinitis. Companion animals serve as reservoirs for several zoonotic species of Bartonella, and may also serve as sentinels for zoonotic Bartonella species harbored by wildlife. This article provides an overview of bartonellosis of dogs and cats, and discusses public health implications of animal bartonellosis. PMID- 20018459 TI - A new definition of early age at onset in alcohol dependence. AB - OBJECTIVE: The accurate cut-off of an early onset of alcohol dependence is unknown. The objectives of this analysis are (1) to confirm that ages at onset variability in alcohol dependence is best described as a two subgroups entity, (2) to define the most appropriate cut-off, and (3) to test the relevancy of such distinction. METHOD: Data were drawn the Epidemiologic Survey on Alcohol and Related Conditions (NESARC). This study focused on the 4782 adults with lifetime alcohol dependence. RESULTS: The best-fit model distinguished two subgroups of age at onset of alcohol dependence, with a cut-off point at 22 years. Subjects with an earlier onset of alcohol dependence (< or = 22 years old) reported higher lifetime rates of specific phobia, antisocial behaviors and nearly all addictive disorders. CONCLUSIONS: The early onset of alcohol dependence is best defined as beginning before the age of 22 years. PMID- 20018463 TI - Two genetically closely related pigeon paramyxovirus type 1 (PPMV-1) variants with identical velogenic fusion protein cleavage sites but with strongly contrasting virulence. AB - Two pathogenetically different pigeon paramyxovirus type 1 (PPMV-1) virus clones were recently derived by passage of a single isolate with an intracerebral pathogenicity index (ICPI) of 0.32. The virus clones had an ICPI of 0.025 and 1.3, respectively (Fuller et al., 2007). Remarkably both viruses contained a cleavage site motif in the precursor fusion (F) protein that is usually associated with virulent viruses. In the current study, both viral genomes were completely sequenced and only four amino acid differences were observed. Of these, two were considered irrelevant on theoretical grounds and two amino acid changes were unique for virus 0.025. The latter were introduced into an infectious clone of a virulent Newcastle disease virus strain, individually and combined, and the effects of the mutations on pathogenicity were examined. The results indicate that only the S453P substitution in the F protein had a modest effect on pathogenicity. We were not able to identify the molecular basis for the pathogenicity difference between both viruses. However, our observations emphasize the need to determine both the virulence (ICPI) and the sequence of the cleavage site of the F protein to avoid dismissing of potential virulent PPMV-1 isolates. PMID- 20018464 TI - Tylosin and chlorotetracycline decrease the duration of fecal shedding of E. coli O157:H7 by swine. AB - Previously we have shown that experimentally infected swine, fed an antibiotic free diet, can become colonized and shed Escherichia coli O157:H7 for at least 2 months. However, in epidemiological studies this organism is only rarely recovered from domestic swine and the basis for this discrepancy is not clear. In this report we demonstrate that significantly fewer pigs fed diets containing subtherapeutic levels of either tylosin or chlorotetracycline shed E. coli O157:H7 for longer than 2 weeks compared to those fed an antibiotic-free diet. In contrast to tylosin and chlorotetracycline, the addition of bacitracin methylene disalicylate to the diet did not influence the recovery of E. coli O157:H7. These results suggest that some antibiotics may alter the gastrointestinal tract flora in ways that create a less favorable environment for E. coli O157:H7 in swine. PMID- 20018465 TI - Molecular and serological analysis of the epidemiology of myxoma virus in rabbits. AB - The epidemiology of myxoma virus was studied by serology and molecular analysis of restriction fragment length polymorphisms (RFLPs) in genomic DNA. 159 isolates of myxoma virus were made over a period of 5 spring/summer epidemics from 12 field sites in south-eastern Australia. Virus isolates were classified into 10 genetic types using RFLPs detected with a panel of nine restriction endonucleases. Between 3 and 6 different genetic types were found during spring/summer periods across all sites and up to 3 different genetic types were isolated during an epidemic on a single site. The predominant type tended to change each year. A widespread mutation was identified in two genetic types with replacement of the 3' two-thirds of the M009L gene at the left hand inverted terminal repeat junction with a duplication of the region containing the M156R, M154L and M153R genes from the right hand end of the genome. This demonstrated how myxoma virus can potentially evolve by expansion of the inverted terminal repeat boundaries. PMID- 20018466 TI - Spermatogenic efficiency in the spiny rat, Trinomys moojeni (Rodentia: Echimyidae). AB - The spiny rat (Trinomys moojeni) is a rodent found in the Atlantic Forest, which is considered one of the most diverse and threatened biomes in the world. Knowledge on reproductive biology and physiology is critical to conservation and species management, allowing the prevention of extinction and the use of males in natural and artificial reproduction programs. The main objectives of the present study were to investigate the testis structure as well as spermatogenic and Sertoli cell efficiency in the spiny rat captured in the Caraca Natural Reserve, a fragment of the Atlantic Forest located in the State of Minas Gerais, Brazil. Ten sexually mature spiny rats were analyzed. Intraperitoneal injections of tritiated thymidine were administered to estimate duration of spermatogenesis. The testes were perfused-fixed in buffered glutaraldehyde and routinely processed for histological and morphometric analyses as well as the characterization of the stages of seminiferous epithelium cycle. Volume density (%) of seminiferous tubules and Leydig cells were 97 +/- 0.3 and 0.3 +/- 0.02, respectively. The duration of one spermatogenic cycle and total duration of spermatogenesis were 8.6 +/- 0.1 and 38.5 +/- 0.5 days, respectively. Due to the very great volume density of the seminiferous tubules, short duration of spermatogenesis, tubule length per gram of testis (approximately 40 m), great Sertoli cell efficiency (approximately 15 spermatids per Sertoli cell) and large number of Sertoli cells per testis gram (53 million), spermatogenic efficiency in the spiny rat (82 million) is by far the greatest of the mammalian species investigated thus far. PMID- 20018467 TI - Hormone replacement after gynaecological cancer. AB - Treatment of gynaecological cancer frequently results in the loss of ovarian function and menopausal symptoms. Symptoms of iatrogenic menopause are usually significantly more intense than those of natural menopause due to sudden onset of symptoms, younger age and its effects on common physical and psychological problems of cancer therapy like body image concerns and sexual dysfunction. The most effective treatment for menopausal symptoms is hormone replacement therapy (HRT). However, it is very controversial if HRT is safe in patients after a gynaecological malignancy. The main concerns are the potential stimulation of residual cancer and the induction of new hormone-dependent disease. However, the majority of the most common gynaecological malignancies like squamous cell carcinomas of the cervix, serous papillary epithelial ovarian carcinomas and squamous cell carcinomas of the vulva are not oestrogen dependent. Furthermore, current scientific evidence does not show HRT to adversely affect the outcome in patients after treatment for hormone sensitive cancers like early stage endometrioid adenocarcinomas of the endometrium. There are only a small number of gynaecological malignancies like low grade endometrial stromal sarcomas in which HRT is an absolute contraindication. Therefore, as maintaining quality of life and minimising the physical and psychological impact of treatment side effects is one of the most important factors in cancer care, it is imperative to give patients unbiased information about their individual cancer which in most cases will allow them to use HRT without any detrimental effect on their survival. PMID- 20018468 TI - Correlates and predictors of self-rated health and ambulatory status among elderly women - Cross-sectional and 10 years population-based cohort study. AB - AIM: To investigate the correlates and predictors of self-rated health (SRH) and self-reported ambulatory status (AMB) among elderly women. SUBJECTS AND METHODS: A random sample of 1620 postmenopausal women aged 53-66 years in 1994 was selected from Osteoporosis Risk Factor and Prevention (OSTPRE) - study cohort in Finland. SRH and AMB were assessed in baseline (1994) and after 10-year follow-up (2004). A prognostic model was developed to predict the probability of good 10 year SRH and AMB. RESULTS: Full AMB was correlated with and predicted by good SRH (OR=4.3, p<0.001). Good quality of life (QoL) was both correlate and predictor of good 10-year SRH (OR=2.0, p<0.001) and full AMB (OR=2.6, p<0.001). Low BMI was associated with good SRH (OR=0.97, p<0.049) and full AMB (OR=0.92, p<0.001), good grip strength with good SRH (OR=2.7, p<0.001) and ability to squat down with full AMB (OR=1.7, p=0.002). Cardiovascular diseases, arthritis and back pain were common significant negative correlates (p<0.001) of baseline AMB and SRH out of variety of chronic diseases with significant association with either one. The significant (p<0.05) baseline predictors in final prognostic model for good 10 year SRH included baseline AMB and SRH, QoL, age, BMI, grip strength and psychiatric diseases (C-index=0.81) and for full 10-year AMB ability to squat, baseline AMB and SRH, QoL, BMI, hypertension and arthritis (C-index=0.81). These risk scores had better predictive power in comparison to that of baseline SRH and AMB alone (p<0.001 in univariate Z-score test). CONCLUSIONS: Among elderly women AMB is strongly linked to SRH. SRH and AMB are affected mostly by musculoskeletal, cardiovascular and psychiatric diseases as well as body anthropometry, muscle strength and subjective QoL. The present study provides a novel algorithm for predicting probability of full AMB and good SRH in postmenopausal women. PMID- 20018469 TI - The Asian Menopause Survey: knowledge, perceptions, hormone treatment and sexual function. AB - OBJECTIVE: To provide current insights into the opinions, attitudes, and knowledge of menopausal women in Asia regarding menopause and hormone replacement therapy (HRT). STUDY DESIGN: Cross-sectional. MAIN OUTCOME MEASURES: Between January 2006 and February 2006, 1000 postmenopausal women from China, Malaysia, Taiwan, Thailand and Hong Kong were interviewed to determine postmenopausal symptoms, HRT use and knowledge, breast discomfort and knowledge of breast cancer risks, and sexual function. RESULTS: Almost all women reported experiencing postmenopausal symptoms. Sleeplessness (42%) was reported as the main reason for seeking treatment. On average, 54% of women were aware of HRT, despite the fact that most (38%) were unable to mention any associated benefits. Most women had used natural or herbal treatments (37%) for the alleviation of menopausal symptoms. Only 19% had received HRT. 27% of respondents reported having breast discomfort, while 70% reported performing self-breast examinations. 53% of women had never received a mammogram, despite breast cancer concern (50%). 24% of women described HRT as being a risk factor for breast cancer. Most women and their partners reported no reductions in sexual function (66 and 51%, respectively), while 90% of respondents did not seek treatment for reduced sexual function. In the event of sexual dysfunction, 33% of women replied that they would be willing to seek treatment. CONCLUSIONS: Many Asian women experience postmenopausal symptoms that are often left untreated (due to the acceptance of menopause as a natural process) or treated with herbal/natural remedies. There was a general lack of knowledge among these women regarding treatment options, HRT, and possible risks associated with HRT. A more concerted effort should be made to better disseminate information regarding the pathogenesis and risk factors associated with breast cancer, menopause, and menopausal symptoms to Asian women. PMID- 20018470 TI - Two cases of lysergamide intoxication by ingestion of seeds from Hawaiian Baby Woodrose. AB - We describe two cases of human consumption of seeds from Argyreia nervosa (Hawaiian Baby Woodrose), which resulted in one fatality due to falling from a building and one surviving witness. The principal psychoactive constituent of the seeds, lysergamide (LSA), was recovered from blood and urine samples by mixed mode cation exchange solid-phase extraction and quantified by ultra performance liquid chromatography-time of flight mass spectrometry (UPLC-ToF/MS). The LSA concentrations were determined by UPLC-ToF/MS to be 4.9 microg/L in blood and 1.0mg/L in urine in the dead person and 1.8 microg/L in blood and 0.50mg/L in urine in the living person. These analytical findings were found to be in accordance with the case story, which indicated that seeds had been ingested and also noted psychological reactions, i.e. the will to jump out of the window. Other findings in the dead person were 22 microg/L THC in blood, 0.71 g/L ethanol in blood and 1.0 g/L ethanol in vitreous humor. Constituents originating from the seeds of A. nervosa, i.e. LSA, ergonovine, lysergic acid alpha-hydroxyethylamide were also identified in the biological samples. The 2-hydroxy-3-oxo metabolites of LSA and ergonovine were identified in the urine sample of the deceased. PMID- 20018471 TI - Insect succession and carrion decomposition in selected forests of Central Europe. Part 2: Composition and residency patterns of carrion fauna. AB - The insect fauna of pig carcasses was monitored in different seasons and forests of Western Poland (Central Europe). The composition of carrion fauna and selected features of residency in carrion in adults and larvae of particular taxa were analysed. A total of 131 adult and 36 larval necrophilous taxa were collected. Only 51 adult species and 24 larval taxa were minimally abundant (>or=10 specimens) at least on one carcass. As for the composition of carrion fauna, there were large differences between seasons, but no important differences between forest types. In most species of Diptera, length of the presence period of adults was between 35 and 65% of the sampling interval, while in most species of Coleoptera, it was above 60%. Only in a few species (e.g., Saprinus semistriatus, Necrodes littoralis or Creophilus maxillosus) was the presence period shorter than 35% of the sampling interval. Interestingly, in some adult Coleoptera (e.g., Necrobia violacea) very long presence periods were recorded. In most taxa, the length of the presence period of larvae was between 40 and 65% of the sampling interval. Only Calliphora vomitoria, Phormia regina, Hydrotaea dentipes, N. littoralis and C. maxillosus had shorter presence periods of larvae. As a rule, residency of adults was broken, whereas residency of larvae was unbroken. Moreover, in adults, two distinct residency patterns were observed; with breaks clumped in the final part of the presence period and with breaks evenly distributed inside the presence period. Almost in all taxa, the time of appearance showed the closest relationship to the onset of bloating. The relationship was significant, positive and strong in adults of P. regina, Fannia manicata, Hydrotaea ignava, Stearibia nigriceps, S. semistriatus, N. littoralis and C. maxillosus as well as larvae of P. regina, H. dentipes, H. ignava, S. nigriceps, N. littoralis, Oiceoptoma thoracicum, Thanatophilus sp., C. maxillosus and Philonthus sp. Interestingly, in some forensically significant taxa (e.g., adults of N. violacea or Thanatophilus rugosus), we found no significant relationship between the time of their appearance and the onset of any decompositional process. Implications for the succession-based post-mortem interval (PMI) estimation, determination of the carcass movement and the season of death are discussed. PMID- 20018472 TI - [Monoarthritis caused by Streptococcus pyogenes after nasal septoplasty: interest of the nasal decontamination in preoperative care]. AB - The nasal septoplasty is a very current intervention in otorhinolaryngology surgery. The infectious complications of this intervention are rare and mostly mild. We report here the case of a patient hospitalized in ambulatory surgery within a fracture of the nose with luxation of the septum in the nasal fossa. This patient was operated for a reduction of this fracture with septoplasty. Twelve hours after the intervention the patient presented septic arthritis due to Streptococcus pyogenes. The tracks of prevention are presented. PMID- 20018473 TI - Identification and characterization of degradation products of irbesartan using LC-MS/TOF, MS(n), on-line H/D exchange and LC-NMR. AB - Irbesartan was subjected to hydrolytic, oxidative, photolytic and thermal stress, according to ICH guideline Q1A (R2). The drug showed degradation only in acidic, basic and photoacidic conditions, while it was stable to other stress conditions. A total of three degradation products were formed, which were separated on a C-8 column employing a gradient HPLC method. Initially, a complete mass fragmentation pathway of the drug was established with the help of MS/TOF, MS(n) and H/D exchange studies. Subsequently, the degradation products were subjected to LC MS/TOF and on-line H/D exchange mass studies to obtain their accurate mass, fragment pattern and number of labile hydrogens. The MS results helped to assign tentative structures to degradation products, which were verified through (1)H and 2D COSY LC-NMR experiments. The products were identified as (2'-(1H-tetrazol 5-yl)biphenyl-4-yl)methanamine, 1-(1-((2'-(1H-tetrazol-5-yl)biphenyl-4 yl)methylamino)pentylideneamino)cyclopentane carboxylic acid and 2-butyl-3 (tetrazolo[1,5-f]phenanthridin-6-ylmethyl)-1,3-diazaspiro[4.4]non-1-en-4-one. The structures were justified by mechanisms of their formation. PMID- 20018474 TI - Study and characterization of crystalline hydrate/polymorph forms of 5,11-dihydro 11-ethyl-5-methyl-8-(2-(1-oxido-4-quinolinyl)ethyl-6H-dipyrido(3,2-B:2',3' E)(1,4)diazepin-6-one by solid-state NMR and solution NMR. AB - A novel inhibitor of reverse transcriptase was studied by solid-state NMR. Three phases of the compound were examined which included the dihydrate and two anhydrous polymorphs (Form I and Form III). By correlating (1)H and (13)C solution NMR with the solid-state (13)C NMR CP/MAS and CPPI spectral editing experiments, comparative (13)C assignments were made for each phase. Polymorphs of Form I and Form III and the dihydrate were easily distinguished based upon chemical shift patterns of the carbon resonances. The (1)H spin-lattice relaxation times were also measured for each phase which provided information on the mobility and relative crystallinity. The (13)C ssNMR spectrum of Form I showed the presence of a minor component identified as the dihydrate. Weight/percent quantitation of major and minor components in Form I was obtained from integrated intensities of a 50:50 mixture containing weighed amounts of Form I and the pure dihydrate. Comparison of the ssNMR and X-ray powder diffraction techniques is discussed. PMID- 20018475 TI - People behind exclusive eponyms of radiologic signs (Part II). PMID- 20018476 TI - Which clinical and demographic factors predict poor insight in individuals with obsessions and/or compulsions? AB - This study examined clinical and demographic predictors of poor insight in individuals with obsessions and/or compulsions. It was hypothesized that a combination of factors would explain variance on two measures of insight. Ninety four individuals with obsessive-compulsive symptoms participated in a cross sectional questionnaire-based design, completing measures relating to obsessions and compulsions, depression and anxiety levels, and demographic and clinical information. Results revealed that although demographic factors did not predict poor insight, clinical and comorbid variables as an overall model did. More comorbidities and ordering compulsions independently predicted poor insight, particularly when accompanied by high anxiety and high severity of compulsions. These findings highlight four clinical factors that could act as indicators of poor insight for clinicians when assessing individuals for therapy, since insight itself is difficult to assess accurately. Limitations and implications for future research are discussed. PMID- 20018477 TI - On the sparseness of 1-norm support vector machines. AB - There is some empirical evidence available showing that 1-norm Support Vector Machines (1-norm SVMs) have good sparseness; however, both how good sparseness 1 norm SVMs can reach and whether they have a sparser representation than that of standard SVMs are not clear. In this paper we take into account the sparseness of 1-norm SVMs. Two upper bounds on the number of nonzero coefficients in the decision function of 1-norm SVMs are presented. First, the number of nonzero coefficients in 1-norm SVMs is at most equal to the number of only the exact support vectors lying on the +1 and -1 discriminating surfaces, while that in standard SVMs is equal to the number of support vectors, which implies that 1 norm SVMs have better sparseness than that of standard SVMs. Second, the number of nonzero coefficients is at most equal to the rank of the sample matrix. A brief review of the geometry of linear programming and the primal steepest edge pricing simplex method are given, which allows us to provide the proof of the two upper bounds and evaluate their tightness by experiments. Experimental results on toy data sets and the UCI data sets illustrate our analysis. PMID- 20018478 TI - Geoepidemiology of COPD and idiopathic pulmonary fibrosis. AB - Progress in improving patient outcomes and advancing therapeutics in chronic obstructive pulmonary disease (COPD) and idiopathic pulmonary fibrosis (IPF) is hampered by phenotypic heterogeneity and variable responsiveness to clinical interventions that are not fully explained by currently held disease paradigms for COPD and IPF. Although these chronic lung diseases differ in their geoepidemiology and immunopathogenesis, emerging evidence suggest that organ specific autoimmunity may underlie subphenotypes of COPD and IPF. In particular, the links to tobacco smoking, diet, gender, and environment are explored in this review. We also highlight potential mechanisms that could guide future investigations in both laboratory and clinical settings. A paradigm shift is needed in how we think about COPD and IPF, based on geoepidemiology and a broader understanding of disease pathogenesis that may ultimately lead to new therapies and improved patient outcomes. PMID- 20018479 TI - Light, including ultraviolet. AB - Ultraviolet (UV) light is intricately linked to the functional status of the cutaneous immune system. In susceptible individuals, UV radiation can ignite pathogenic inflammatory pathways leading to allergy or autoimmunity. In others, this same UV radiation can be used as a phototherapy to suppress pathogenic cutaneous immune responses. These vastly different properties are a direct result of UV light's ability to ionize molecules in the skin and thereby chemically alter them. Sometimes these UV-induced chemical reactions are essential, the formation of pre-vitamin D(3) from 7-dehydrocholesterol, for example. In other instances they can be potentially detrimental. UV radiation can ionize a cell's DNA causing adjacent pyrimidine bases to chemically bond to each other. To prevent malignant transformation, a cell may respond to this UV-induced DNA damage by undergoing apoptosis. Although this pathway prevents skin cancer it also has the potential of inducing or exacerbating autoreactive immune responses by exposing the cell's nuclear antigens. Ultraviolet-induced chemical reactions can activate the immune system by a variety of other mechanisms as well. In response to UV irradiation keratinocytes secrete cytokines and chemokines, which activate and recruit leukocytes to the skin. In some individuals UV-induced chemical reactions can synthesize novel antigens resulting in a photoallergy. Alternatively, photosensitizing molecules can damage cells by initiating sunburn like phototoxic reactions. Herein we review all types of UV-induced skin reactions, especially those involving the immune system. PMID- 20018480 TI - Metabolic origin of hypovitaminosis C in acutely hospitalized patients. AB - OBJECTIVE: Recent studies have indicated a high prevalence of hypovitaminosis C in acutely hospitalized patients. It is unclear whether hypovitaminosis C in this setting represents deficiency or tissue redistribution of the vitamin as part of the acute-phase response. METHODS: We administered vitamin C for 1 wk to acutely hospitalized, but not critically ill patients with hypovitaminosis C, on the assumption that a large increase in plasma and mononuclear leukocyte vitamin C concentrations, a decrease in metabolic markers of oxidative stress, or an improvement in psychologic mood state would implicate the initial condition as nutritional deficiency rather than tissue redistribution. RESULTS: Vitamin C administration increased plasma and mononuclear leukocyte vitamin C concentrations from subnormal (16.3 +/- 12.4 MUmol/L and 6.5 +/- 5.5 mmol/L, respectively) to normal (71.0 +/- 30.9 MUmol/L, P < 0.0001, and 8.2 +/- 6.8 mmol/L, P < 0.015); the mood disturbance score improved by 33% (P < 0.008). There was no increase in plasma glutathione concentrations or a reduction in plasma or mononuclear leukocyte malondialdehyde concentrations. An inverse relation was observed between plasma C-reactive protein and plasma vitamin C concentrations (P = 0.006). CONCLUSION: Although associated with systemic inflammation, the metabolic features of hypovitaminosis C in acutely hospitalized, non-critically ill patients are more consistent with deficiency than with tissue redistribution. PMID- 20018481 TI - Influence of traumatic brain injury on potassium and phosphorus homeostasis in critically ill multiple trauma patients. AB - OBJECTIVE: The intent of this study was to ascertain whether multiple trauma patients with traumatic brain injury (TBI) had lower serum concentrations of potassium and phosphorus and required more aggressive supplementation than multiple trauma patients without TBI. METHODS: Ventilator-dependent adult patients without renal impairment who were admitted to the trauma intensive care unit or neurosurgical intensive care unit and who received enteral nutrition were evaluated for the first 14 d after hospital admission. Patients were grouped according to the presence or absence of TBI. Target serum concentrations for potassium and phosphorus were 4 mEq/L and 4 mg/dL, respectively. Electrolyte repletion therapy was given according to the nutritional support service guidelines. RESULTS: Fifty trauma patients (25 with and without TBI) were studied. Daily serum potassium concentrations were consistently lower for those with TBI (P < or = 0.001), whereas the mean net potassium intake was greater (1.3 +/- 0.5 versus 0.7 +/- 0.3 mEq x kg(-1) x d(-1), respectively, P < or = 0.001). Serial serum phosphorus concentrations were similar between groups (P = NS) except for a significantly lower serum phosphorus concentration for trauma patients with TBI on day 3 after hospital admission (2.5 +/- 0.5 versus 2.9 +/- 0.7 mg/dL, respectively, P < or = 0.05). However, the mean net phosphorus intake was significantly greater for trauma patients with TBI (0.65 +/- 0.25 versus 0.45 +/- 0.17 mmol x kg(-1) x d(-1), P < or = 0.001). CONCLUSION: Potassium and phosphorus requirements are greater for multiple trauma patients with TBI compared with those without TBI. PMID- 20018482 TI - Effect of nutritional support team restructuring on the use of parenteral nutrition. AB - OBJECTIVE: The inappropriate use of parenteral nutrition (PN) continues to be a problem, despite several decades of efforts to improve the situation. We restructured our existing nutritional support team employing methods involving the institution's systems and individual physician interaction and education. Our aim was to study the effects of these changes on the use of PN in our institution. METHODS: Records of all non-critically ill patients seen by our nutritional support team for PN during 2003-2004 (phase I, before restructuring) and 2005-2006 (phase II) were reviewed. Patients were classified under "appropriate," "inappropriate," and "marginal" categories based on published guidelines. During phase II, a new committee was formed, policies and procedures were updated, educational activities were increased, emphasis was placed on evidence-based guidelines, and periodic bedside rounds with the team physicians were initiated. Communication with referring physicians was improved. RESULTS: Three hundred three of 335 patients in phase I and 271 of 333 patients in phase II were eligible for inclusion in analysis. Appropriate PN increased from 71.3% to 83.4%; inappropriate PN decreased from 16.5% to 8.9% (P = 0.002). CONCLUSION: Restructuring of the nutritional support team improved the proper utilization of PN and decreased inappropriate use of PN in a public teaching hospital. PMID- 20018483 TI - Safety and tolerance of the human milk probiotic strain Lactobacillus salivarius CECT5713 in 6-month-old children. AB - OBJECTIVE: Intestinal microbiota plays an important role in the prevention of certain diseases during the pediatric years. Thus, there is an increasing interest in the addition of probiotics to infant formulas. The aim of this study was to evaluate the safety of a follow-on formula with Lactobacillus salivarius CECT5713 in 6-mo-old children. METHODS: The antibiotic susceptibility of L. salivarius CECT5713 was analyzed by a dilution method. A double-blinded, randomized, placebo controlled study was performed. Children (n = 80) were distributed in two groups and consumed the formula supplemented or not with probiotics (2 * 10(6) colony-forming units [cfu]/g) during 6 mo. Fecal samples were collected at enrollment, at 3 mo, and at the end of trial. Clinical and anthropometric evaluations were performed. Depending on the variable, one-way or two-way repeated measures analysis of variance were used for the statistical analysis. RESULTS: The antibiotic susceptibility profile of the strain resulted as safe. No adverse effects associated with the consumption of the probiotic formula were reported. In addition, clinical parameters did not differ between groups. Consumption of the probiotic supplemented formula led to an increase in the fecal lactobacilli content (7.6 +/- 0.2 versus 7.9 +/- 0.1 log cfu/g, P < 0.05). Lactobacillus salivarius CECT5713 was detected in the feces of volunteers from the probiotic group. Probiotic consumption induced a significant increase in the fecal concentration of butyric acid at 6 mo. CONCLUSION: Thus, a follow-on formula with L. salivarius CECT5713 is safe and well tolerated in 6-mo-old infants. PMID- 20018484 TI - Malnutrition and pressure ulcer risk in adults in Australian health care facilities. AB - OBJECTIVE: To determine the effect of nutritional status on the presence and severity of pressure ulcer. METHODS: A multicenter, cross-sectional audit of nutritional status of a convenience sample of subjects was carried out as part of a large audit of pressure ulcers in a sample of Queensland, Australia, public healthcare facilities in 2002 and 2003. Dietitians in 20 hospitals and 6 residential aged care facilities conducted single-day nutritional status audits of 2208 acute and 839 aged care subjects using the Subjective Global Assessment. The effect of nutritional status on the presence and severity (highest stage and number of pressure ulcers) was determined by logistic regression in a model controlling for age, gender, medical specialty, and facility location. The potential clustering effect of facility was accounted for in the model using an analysis of correlated data approach. RESULTS: Subjects with malnutrition had adjusted odds ratios of 2.6 (95% confidence interval 1.8-3.5, P<0.001) of having a pressure ulcer in acute care facilities and 2.0 (95% confidence interval 1.5 2.7, P<0.001) for residential aged care facilities. There was also increased odds ratio of having a pressure ulcer, and having a more severe pressure ulcer (higher stage pressure ulcer and/or a higher number) with increased severity of malnutrition. CONCLUSION: Malnutrition was associated with at least twice the odds ratio of having a pressure ulcer of in public health care facilities in Queensland. Action must be taken to identify, prevent, and treat malnutrition, especially in patients at risk of pressure ulcer. PMID- 20018485 TI - Maternal malnutrition during lactation affects folliculogenesis, gonadotropins, and leptin receptors in adult rats. AB - OBJECTIVE: The goal of this study was to evaluate if maternal malnutrition during lactation could possibly program folliculogenesis, the ovarian expression of gonadotropins, leptin, and their receptors. METHODS: At parturition, dams were randomly assigned to a control group (C), with free access to a standard laboratory diet containing 23% protein, and a protein-energy-restricted group (PER), with free access to an iso-energy and protein-restricted diet containing 8% protein. After weaning, all female pups had free access to the standard laboratory diet until 90 d of age when they were euthanized in the diestrum stage. RESULTS: Maternal malnutrition caused decreases in the number of primordial (C 6.60 +/- 0.24, PER 5.20 +/- 0.20, P = 0.01), primary (C 5.80 +/- 0.66, PER 4.00 +/- 0.31, P = 0.04), and Graafian (C 2.18 +/- 0.29, PER 1.08 +/- 0.37, P = 0.05) follicle numbers. Maternal malnutrition led to a significant decrease in the aromatase mRNA expression (C 0.536 +/- 0.008, PER 0.353 +/- 0.041, P = 0.01) follicle-stimulating hormone receptor (C 1.25 +/- 0.17, PER 0.75 +/- 0.02, P = 0.04), luteinizing hormone receptor (C 0.93 +/- 0.09, PER 0.54 +/- 0.10, P = 0.03), leptin (C 0.55 +/- 0.03, PER 0.42 +/- 0.03, P = 0.04), Ob-R (C 1.05 +/- 0.12, PER 0.64 +/- 0.07, P = 0.03), and Ob-Rb (C 1.34 +/- 0.21, PER 0.47 +/- 0.10, P = 0.02) transcripts when compared with C. CONCLUSION: Maternal malnutrition during lactation modulates folliculogenesis and the expression of the different isoforms of leptin and gonadotropin receptors and the aromatase enzyme. This probably is a consequence of alterations in perinatal leptin concentrations that may play a crucial role in determining the occurrence of long term metabolic changes. PMID- 20018487 TI - Enteral nutrition for severe malnutrition in chronic intestinal pseudo obstruction. AB - OBJECTIVE: Chronic intestinal pseudo-obstruction (CIPO) is a rare intestinal motility disorder. A prolonged avoidance of food due to fear of aggravation of postprandial symptoms leads to severe malnutrition. We report a case of a 21 y old man who was diagnosed as CIPO with a history of recurrent intestinal colic and obstructive symptoms, slow transit type of constipation, bilateral hydronephrosis (non-obstructive), motor dysphagia without any evidence of demonstrable mechanical obstruction. Our aim was to keep his post prandial symptoms to a minimum and nutritionally build him up with enteral nutrition (EN). METHODS: He had life threatening malnutrition (BMI of 11 kg/m(2)) and significant postprandial distension with an intake more than 100 ml, compromising the quality of life. In view of a normal absorptive function of the gut, TPN was ruled out and the patient was treated with enteral nutrition (oral & tube) only. The EN regimen followed was ad libitum oral intake along with nocturnal NG tube feeding. Initially a full strength semi-elemental formula at 50 ml/hour was given, later shifted to polymeric formula at 100 ml/hour. Serum levels of magnesium, phosphate and potassium were regularly monitored to prevent refeeding syndrome. He ws constantly motivated, counseled and monitored. RESULT: With a gradual increase in the intake from 300 Kcal to 1400 Kcal he was discharged. Eight months from discharge he had a weight of 58 kg (BMI = 22.3 kg/m(2)), with resumption of normal activities and marked improvement in the quality of life. CONCLUSION: Carefully planned EN along with motivation, psychological support and regular monitoring are the keys to nutritional management in CIPO. PMID- 20018486 TI - Obestatin and ghrelin interplay in hemodialysis patients. AB - OBJECTIVE: Compounds involved in the regulation of appetite and body composition appear to be of interest in chronic kidney disease. The purpose of this study was to analyze plasma obestatin and acyl and des-acyl ghrelin in patients on hemodialysis (HD). METHODS: Fifty patients on HD (56.0% women, mean age 62.2 +/- 15.2 y) were studied. Blood samples were collected during fasting, before a regular HD session. Serum acyl and des-acyl ghrelin levels, leptin, and obestatin were measured using enzyme immunometric assay methods. Anthropometric parameters, appetite score, and food intake were recorded. RESULTS: Patients showed elevated serum leptin (34.1 +/- 30 ng/mL), normal acyl ghrelin (137 +/- 116.5 pg/mL), high des-acyl ghrelin (670 +/- 479 pg/mL), and low obestatin (2.0 +/- 1.4 ng/mL) levels compared with healthy volunteers. According to body mass index (BMI), patients with a BMI >23 kg/m(2) had significantly lower plasma obestatin. In contrast, leptin levels were increased and acyl ghrelin tended to be higher in these patients. There was a strong positive correlation between obestatin and des acyl ghrelin (r = 0.56, P = 0.0001) and inverse correlations between obestatin and BMI (r = -0.40, P = 0.007), waist circumference (r = -0.38, P = 0.024), and C reactive protein (r = -0.29, P = 0.048). By multivariate analysis, obestatin was independently and positively correlated with des-acyl ghrelin (P = 0.01), but not with C-reactive protein, BMI, or waist circumference. CONCLUSION: In summary, patients on HD exhibited increased plasma levels of des-acyl ghrelin, normal acyl ghrelin levels, and low obestatin levels. In lean patients, the obestatin and des acyl ghrelin levels were increased, suggesting that these hormones may influence appetite and body composition in patients on HD. PMID- 20018488 TI - Effect of Brazil nut supplementation on the blood levels of selenium and glutathione peroxidase in hemodialysis patients. AB - OBJECTIVE: In patients who have undergone hemodialysis, large amounts of reactive oxygen species (ROS) are produced and, at higher concentrations, ROS are thought to be involved in the pathogenesis of cardiovascular disease. It has been proposed that selenium (Se) may exert an antiatherogenic influence by reducing oxidative stress. The richest known food source of selenium is the Brazil nut (Bertholletia excelsa, family Lecythidaceae), found in the Amazon region. We evaluated the effect of Brazil nut supplementation on blood levels of Se and glutathione peroxidase (GSH-Px) activity in patients on hemodialysis. METHODS: A total of 81 patients on hemodialysis (52.0+/-15.2 y old, average time on dialysis 82.3+/-91.4 mo, body mass index 24.9+/-4.4 kg/m(2)) from the RenalCor and RenalVida Clinics in Rio de Janeiro, Brazil, were studied. All patients received one nut (around 5 g, averaging 58.1 MUg Se/g) a day for 3 mo. The Se concentrations in the nuts and in plasma and erythrocytes were determined by atomic absorption spectrophotometry with hydride generation (Hitachi, Z-500). GSH Px levels were measured using Randox commercial kits. RESULTS: Plasma Se (18.8+/ 17.4 MUg/L) and erythrocyte (72.4+/-37.9 MUg/L) levels were below the normal range before nut supplementation. After supplementation, the plasma level increased to 104.0+/-65.0 MUg/L and erythrocytes to 244.1+/-119.5 MUg/L (P<0.0001). The activity of GSH-Px also increased after supplementation, from 46.6+/-14.9 to 55.9+/-23.6 U/g of hemoglobin (P<0.0001). Before supplementation, 11% of patients had GSH-Px activity below the normal range (27.5-73.6 U/g of hemoglobin). After supplementation, all patients showed GSH-Px activity within the normal range. CONCLUSION: The data revealed that the investigated patients presented Se deficiency and that the consumption of only one Brazil nut a day (5 g) during 3 mo was effective to increase the Se concentration and GSH-Px activity in these patients, thus improving their antioxidant status. PMID- 20018489 TI - A technique for functional and aesthetic reconstruction of the oral commissure and buccal mucosa. AB - Oral mucosal lesions such as hyperkeratosis can enlarge and involve adjacent anatomic structures. Surgical treatment may cause significant morbidity in terms of loss of function due to scarring and compromised aesthetics due to failure to restore the original muscle anatomy and overlying soft tissue. The authors present a case of hyperkeratosis of the oral commissure and buccal mucosa, which was successfully reconstructed by using vermilion myomucosal flap and A-T advancement. PMID- 20018490 TI - Incidence of venous thromboembolism in oral and maxillofacial surgery: a retrospective analysis. AB - This study retrospectively investigates the incidence and risk factors for venous thromboembolism (VTE) in patients undergoing maxillofacial surgery. Data were obtained from patients treated between January 2005 and June 2006. Patients' records were reviewed for complaints and information relating to deep venous thrombosis (DVT) and pulmonary embolism (PE). All patients completed a questionnaire on complaints relating to DVT and PE. The patients were classified according to risk and the incidence of reported DVT and PE was calculated. The study population comprised 411 patients with a mean age of 32 (+/- 17) years. The median postoperative follow-up period was 53 (23-639) days. None of the patients received thromboembolism prophylaxis. The incidence of VTE in the study group was 0.5%. Logistic regression analysis demonstrated a relationship between body mass index and hospital stay with thromboembolism (p<0.05). Patients undergoing pre implant surgery with bone graft proved to be at risk (p=0.07). The incidence of VTE in oral and maxillofacial surgery seems to be low, but thromboembolism prophylaxis may be justified in patients with clear potential risk factors. PMID- 20018491 TI - Abnormal serotonin receptor expression in DBA/2 mice associated with susceptibility to sudden death due to respiratory arrest. AB - Previous studies indicate that DBA/2 mice may be a useful model of human sudden unexpected death in epilepsy (SUDEP), since these mice exhibit generalized convulsive seizures followed by respiratory arrest (RA). Respiratory deficits, following generalized convulsive seizures, are observed prior to SUDEP in patients. RA that occurs in DBA/2 mice following sound-induced seizures can be prevented by treatments that activate serotonin (5-HT) receptors. 5-HT receptor subtypes in brainstem respiratory centers are important in regulating normal respiration. The present study compared the expression of 5-HT subtype receptor proteins in excised brainstem tissue, containing the rostral ventral medulla respiratory region in DBA/2 mice vs. seizure-resistant C57BL/6J mice, using Western blot analysis. The results indicate that expression of specific 5-HT(2C), 5-HT(3), and 5-HT(4) receptor proteins in the brainstem tissue of DBA/2 mice is significantly diminished, while expression of the 5-HT(2B) receptors is significantly enhanced as compared to C57BL/6J mice. No difference in expression of 5-HT transporter protein is seen. These findings suggest that the DBA/2 mice are susceptible to RA, in part, because of the altered expression of 5-HT receptors. Preliminary studies indicate that 5-HT(2C) receptors may be particularly important, since a 5-HT(2C) agonist is very effective in blocking RA in DBA/2 mice. PMID- 20018492 TI - First-dose and steady-state population pharmacokinetics and pharmacodynamics of piperacillin by continuous or intermittent dosing in critically ill patients with sepsis. AB - The objectives of this study were (i) to compare the plasma concentration-time profiles for first-dose and steady-state piperacillin administered by intermittent or continuous dosing to critically ill patients with sepsis and (ii) to use population pharmacokinetics to perform Monte Carlo dosing simulations in order to assess the probability of target attainment (PTA) by minimum inhibitory concentration (MIC) for different piperacillin dosing regimens against bacterial pathogens commonly encountered in critical care units. Plasma samples were collected on Days 1 and 2 of therapy in 16 critically ill patients, with 8 patients receiving intermittent bolus dosing and 8 patients receiving continuous infusion of piperacillin (administered with tazobactam). A population pharmacokinetic model was developed using NONMEM, which found that a two compartment population pharmacokinetic model best described the data. Total body weight was found to be correlated with drug clearance and was included in the final model. In addition, 2000 critically ill patients were simulated for pharmacodynamic evaluation of PTA by MIC [free (unbound) concentration maintained above the MIC for 50% of the dosing interval (50% f(T>MIC))] and it was found that continuous infusion maintained superior free piperacillin concentrations compared with bolus administration across the dosing interval. Dosing simulations showed that administration of 16g/day by continuous infusion vs. bolus dosing (4g every 6h) provided superior achievement of the pharmacodynamic endpoint (PTA by MIC) at 93% and 53%, respectively. These data suggest that administration of piperacillin by continuous infusion, with a loading dose, both for first dose and for subsequent dosing achieves superior pharmacodynamic targets compared with conventional bolus dosing. PMID- 20018493 TI - Development of the 2nd generation neurokinin-1 receptor antagonist LY686017 for social anxiety disorder. AB - The neurokinin-1 (NK-1) antagonist LY686017 showed activity in preclinical anxiety models. The clinical development of LY686017 included a PET study and a proof-of-concept in social anxiety disorder (SAD). [(11)C]GR205171 was used healthy volunteers receiving 1-100mg/d LY686017 for 28 days to determine brain receptor occupancy (RO). The mean NK-1 RO increased ranged from 25% with 1mg to 93% with 100mg. Subsequently, a 12-week randomized clinical trial tested LY686017 vs. paroxetine, or placebo in SAD. Pharmacokinetic (PK)/RO modeling based on the PET results predicted that once daily dosing of >30mg LY686017 led to sustained trough RO of over 80%. 189 outpatients(1) suffering from SAD were randomly assigned to 12-weeks treatment with 50mg/d LY686017 (N=77), placebo (N=74), or 20mg/d paroxetine (N=38). There was no significant difference between LY686017 and placebo as measured with the Liebowitz Social Anxiety scale (LSAS). The active comparator paroxetine showed positive trends on primary and secondary measures. The plasma concentrations were above the level expected to produce maximal brain NK-1 RO based on the PK/RO relationship obtained in the human PET investigation. Thus, further evaluation of LY686017 for the treatment of SAD does not seem warranted. PMID- 20018494 TI - [Diffuse intrinsic brain stem glioma in children: current treatment and future directions]. AB - Despite the numerous clinical trials undertaken, the prognosis of children with diffuse brain stem glioma remains very poor. This review examines the different strategies for the treatment of malignant brain stem glioma such as radiation therapy, concurrent radiochemotherapy, and classical cytotoxic drugs, with a particular focus on the novel targeted and antiangiogenic drugs recently introduced in pediatric oncology. The strategy using integrin inhibitors is discussed. PMID- 20018495 TI - [Healthcare of imprisoned juveniles: new regulations]. AB - Several new French regulations have come into effect to regulate the healthcare of juvenile offenders in prison with the creation of French Young Offender Institutions. They complete the French prison healthcare methodological guide. This article presents the new developments in the healthcare of juveniles in prison. It specifies the limitations placed on the healthcare team's interventions on imprisoned juveniles. Promoting an individualized prisoner program, as is done in the school context, outlining parental involvement in this program, and withdrawing from the healthcare methodological guide the tasks that are not within the realm of the physician caring for the minor would be measures to ensure good ethical medical practices in prison. These could be applied to French secure training centers and secure children's homes. PMID- 20018496 TI - [Deaths in children with acute dehydration: learning from a nationwide medical insurance company database]. PMID- 20018497 TI - [First diagnosis of cystic fibrosis in Afghanistan and description of a new mutation]. PMID- 20018498 TI - [Developmental amnesia in the premature infant]. AB - All types of memory disorders have been observed in children, although these reports are rare. Developmental amnesia selectively involves episodic daily life memory while semantic learning is respected and general intelligence is not affected. Daily life is severely disturbed by this cognitive disorder usually occurring after hypoxic ischemic injury with bilateral hippocampal atrophy on MRI. Memory disorders are underdiagnosed in at-risk patients and rarely reported. We report on a former small-for-gestational-age preterm infant with no obvious hypoxic event during perinatal life. The follow-up was normal until elementary school. He had to spend 2 years in 1st grade and exhibited some behavioral troubles. At the age of 9, he was suspected of suffering from dyspraxia and was referred to a pediatrics rehabilitation center. IQ and neuropsychological tests were administered and showed selective autobiographical memory impairment defining developmental amnesia. Despite a typical clinical presentation, brain MRI was normal, including the hippocampal area. This observation underlines the need for a prolonged follow-up until school age to assess the outcome of preterm infants. Otherwise, the evaluation will be limited to motor impairment. Particular attention should be paid to memory during the follow-up to avoid misdiagnoses and to plan and adapt these children's educational strategies. PMID- 20018499 TI - Two vicariant Semispathidium species from tropical Africa and central Europe: S. fraterculum nov. spec. and S. pulchrum nov. spec. (Ciliophora, Haptorida). AB - Using standard methods, we describe two new Semispathidium species from semiterrestrial habitats of tropical Africa and central Europe. Semispathidium fraterculum nov. spec. and S. pulchrum nov. spec. differ from each other and from two Namibian (Southwest African) congeners by body size, the shape and location of the extrusomes; the shape of the macronucleus, the number of ciliary rows, the number of cilia within the rows, and the number of dikinetids comprising the dorsal brush rows. The globular resting cyst of S. fraterculum is unique in having countless granules on the inside of the external layer, forming sharp angled rows. The distribution of these and two further, not yet described south African species indicates that the genus Semispathidium evolved in subtropical Africa. Semispathidium fraterculum and S. pulchrum are highly similar, both morphologically and ecologically, differing in mainly one important feature: the shape and location of the extrusomes used for prey capture. This suggests that they evolved from a common ancestor whose areal was divided by a vicariant event, causing an independent evolution of the split populations. We argue that this and other species pairs indicate that the vicariance speciation model is applicable to protists. PMID- 20018500 TI - Learning from HIV epidemics among injecting drug users. AB - HIV/AIDS was the defining issue for international harm reduction during its first twenty years. This issue was marked by strong contrasts: rapid HIV transmission in some populations of injecting drug users, and close to elimination of HIV in other populations; a formidable research base for designing effective HIV programmes and persistent political problems in implementing evidence-based programmes on a public health scale. Elevated rates of HIV infection among ethnic minority drug users have occurred in many different countries. We do not yet have systematic knowledge of how to reduce stigmatization of AIDS or people who use drugs. Nevertheless, international harm reduction for people who use drugs has moved beyond HIV/AIDS to a variety of other health and social problems, while retaining firm bases in science and human rights. PMID- 20018501 TI - Addition of tracers into the polypropylene in view of automatic sorting of plastic wastes using X-ray fluorescence spectrometry. AB - This study focused on the detection of rare earth oxides, used as tracers for the identification of polymer materials, using XRF (X-ray fluorescence) spectrometry. The tests were carried out in a test system device which allows the collection of static measurements of the samples' spectrum through the use of energy dispersive X-ray fluorescence technology. A sorting process based on tracers added into the polymer matrix is proposed in order to increase sorting selectivity of polypropylene during end-of-life recycling. Tracers consist of systems formed by one or by several substances dispersed into a material, to add a selective property to it, with the aim of improving the efficiency of sorting and high speed identification. Several samples containing rare earth oxides (Y(2)O(3), CeO(2), Nd(2)O(3), Gd(2)O(3), Dy(2)O(3), Er(2)O(3) and Yb(2)O(3)) in different concentrations were prepared in order to analyse some of the parameters which can influence the detection, such as the concentration of tracers, the acquisition time and the possible overlapping among the tracers. This work shows that by using the XRF test system device, it was possible to detect 5 of the 7 tracers tested for 1min exposure time and at a concentration level of 1000ppm. These two parameters will play an important role in the development of an industrial device, which indicates the necessity of further works that needs to be conducted in order to reduce them. PMID- 20018502 TI - Technoeconomic aspects of alternative municipal solid wastes treatment methods. AB - This paper considers selected treatment technologies for comingled domestic and similar wastes and provides technoeconomic data and information, useful for the development of strategic management plans. For this purpose, treatment technologies of interest are reviewed and representative flow diagrams, along with material and energy balances, are presented for the typical composition of wastes in Greece; possible difficulties in the use of treatment products, along with their management implications, are discussed, and; cost functions are developed, allowing assessment of the initial capital investment and annual operating costs. Based on the latter, cost functions are developed for predicting the normalized treatment costs of alternative methods (in euro/t of MSW treated), as function of the quantity of MSW processed by plants built and operated (a) by municipality associations, and (b) by private enterprises. Finally, the alternative technologies considered are evaluated on the basis of their cost aspects, product utilization and compatibility with the EU waste framework Directive 2008/98. PMID- 20018503 TI - HMGB1 attenuates anti-metastatic defence of the liver in colorectal cancer. AB - High mobility group box (HMGB) 1 induces apoptosis of monocyte-lineage cells. We examined the effect of HMGB1 on Kupffer cells (KCs). In 50 Dukes C and 12 liver metastasised Dukes D colorectal cancers (CRCs), higher HMGB1 concentration in the primary tumours and metastatic foci, and fewer KCs were found in Dukes D cases than in Dukes C cases. The portal blood HMGB1 concentration was higher in Dukes D cases than in Dukes C cases. HMGB1 induced growth inhibition and apoptosis in mouse KCs in a dose-dependent manner, which was associated with the phosphorylation of c-Jun N-terminal kinase (JNK). JNK inhibition and knockdown of HMGB1 receptor abrogated growth inhibition and apoptosis. In a nude mouse liver metastasis model, the caecal administration of HMGB1 decreased the number of KCs and increased the embedment of Colo320 CRC cells in a dose-dependent manner. HMGB1 transfection increased the liver metastasis of Colo320 cells, and the metastasis was inhibited by anti-HMGB1 antibody administration. These results suggest that HMGB1 secreted from primary tumours decreases the number of KCs and attenuates the anti-metastatic defence of the liver in patients with CRCs. PMID- 20018504 TI - Important perspectives on perioperative pain in elderly patients with cancer. Preface. PMID- 20018505 TI - Lignocellulosic ethanol in India: Prospects, challenges and feedstock availability. AB - India has a pressing need for renewable transportation fuels and bio-ethanol is considered as one of the most important options. Currently the country mandates use of 5% ethanol blending in motor gasoline in several states. The ethanol for this is mainly sourced from molasses feedstock, but this is barely sufficient to meet the current demand. Lignocellulosic biomass is the alternative but the availability of this resource is poorly documented. Also the technologies for ethanol production from lignocellulosic biomass are under preliminary stages of development which warrants extensive R&D in this field. The review discusses the current status of molasses based ethanol production in India and its limitations, the state of technologies for second generation ethanol production and the availability of feedstock for bio-ethanol production. PMID- 20018506 TI - Effects of dilute acid pretreatment conditions on enzymatic hydrolysis monomer and oligomer sugar yields for aspen, balsam, and switchgrass. AB - The effects of dilute acid hydrolysis conditions were investigated on total sugar (glucose and xylose) yields after enzymatic hydrolysis with additional analyses on glucose and xylose monomer and oligomer yields from the individual hydrolysis steps for aspen (a hardwood), balsam (a softwood), and switchgrass (a herbaceous energy crop). The results of this study, in the form of measured versus theoretical yields and a severity analysis, show that for aspen and balsam, high dilute acid hydrolysis xylose yields were obtainable at all acid concentrations (0.25-0.75 wt.%) and temperatures (150-175 degrees C) studied as long as reaction time was optimized. Switchgrass shows a relatively stronger dependence on dilute acid hydrolysis acid concentration due to its higher neutralizing mineral content. Maximum total sugar (xylose and glucose; monomer plus oligomer) yields post-enzymatic hydrolysis for aspen, balsam, and switchgrass, were 88.3%, 21.2%, and 97.6%, respectively. In general, highest yields of total sugars (xylose and glucose; monomer plus oligomer) were achieved at combined severity parameter values (log CS) between 2.20 and 2.40 for the biomass species studied. PMID- 20018507 TI - Continuous microwave-assisted isoflavone extraction system: design and performance evaluation. AB - The purpose of this research was to design, test, and optimize a continuous microwave extraction method using temperature and residence time during and after microwave exposure as optimizing parameters for extraction of major isoflavones (genistin, genistein, daidzin, and daidzein) from soy flour. The extraction yield of four isoflavones at different heating temperatures (55 and 73 degrees C) and extraction times (0, 4, 8, 12, and 16 min) were investigated and compared with yields provided by a conventional solvent extraction method. The microwave prototype consisted of multiple, commercially available, batch-type, house-hold microwave units placed on top of each other in series to impart a continuous operation. The optimum parameters for microwave-assisted extraction of isoflavones were 73 degrees C for 8 min using a 3:1 ethanol-to soy flour ratio. At these parameters, the total yield of isoflavones extracted doubled, while the amount of oil extracted was 12%. Continuous microwave-assisted solvent extraction is a viable method for extraction of soybean isoflavones at relatively short residence times and high throughput. PMID- 20018508 TI - Contributions to a better comprehension of redox-mediated decolouration and detoxification of azo dyes by a laccase produced by Streptomyces cyaneus CECT 3335. AB - The ability of a laccase (EC 1.10.3.2) produced by Streptomyces cyaneus CECT 3335 to decolourise and detoxify azo dyes was assessed. Results showed that a colour loss of 90% was achieved only in the presence of acetosyringone (0.1mM) acting as a redox mediator for the laccase. Toxicological analysis of the decolourised dyes revealed that there was no direct correlation between decolouration and detoxification; in fact, in the case of the dyes Methyl Orange and Orange II, a significant increase in toxicity was produced after the treatment. In contrast, a significant decrease in toxicity was observed after the decolouration of New Coccine and Chromotrope 2R. Finally, HPLC analysis of the dyes after treatment revealed the complete disappearance of both dyes and mediator and a concomitant appearance of new chromatographic peaks which could be responsible of the residual toxicity detected in some cases. PMID- 20018509 TI - Synthesis of novel 7-imino-2-thioxo-3,7-dihydro-2H-thiazolo [4,5-d] pyrimidine derivatives as adenosine A2A receptor antagonists. AB - Novel bicyclic thiazolopyrimidine compounds (15-26) were synthesized to develop adenosine A(2A) receptor (A(2A)R) antagonist for the treatment of Parkinson's disease (PD). The binding affinity of the compounds (15-26) with A(2A)R was evaluated using radioligand binding assay on isolated membranes from stably transfected HEK293 cells. Selectivity of the compounds towards A(2A)R was assessed by comparing their binding affinities with A(1) receptors (A(1)R). cAMP concentrations were measured from HEK293 cells treated with compounds (15-26) as compared to NECA (A(2A)R agonist). The compound (16) possessed strongest A(2A)R binding affinity (K(i) value=0.0038 nM) and selectivity (737-fold) versus A(1)R. Decrease in A(2A)R-coupled release of endogenous cAMP from HEK293 cells treated with compounds (15-26) is evocative of their potential as A(2A)R antagonist. PMID- 20018510 TI - Diaryl piperidines as CB1 receptor antagonists. AB - The syntheses and SAR investigations of novel CB(1) receptor antagonists based on a 1,2-diaryl piperidine core have been described. Optimization of this core afforded a compound with robust in vivo potency by reducing food intake in a mouse DIO model. PMID- 20018511 TI - A novel mitochondrial MTND5 frameshift mutation causing isolated complex I deficiency, renal failure and myopathy. AB - Isolated complex I deficiency is the most commonly reported enzyme defect in paediatric mitochondrial disorders, and may arise due to mutations in nuclear encoded structural or assembly genes, or the mitochondrial genome. We present the clinical, biochemical and molecular genetic data in a young girl whose clinical picture is dominated by chronic renal failure, myopathy and persistent lactic acidosis. An isolated complex I deficiency in muscle was identified due to a novel mutation (m.12425delA) in the MTND5 gene. This single nucleotide deletion is heteroplasmic and detectable in several tissues from the proband but not her mother, suggesting a de novo mutation event. The description of the first frameshift mutation in a mitochondrial complex I gene affirms mitochondrial DNA mutations as an important cause of isolated complex I deficiency in children and the importance of whole mitochondrial genome sequencing in the diagnostic work-up to elucidate the underlying molecular genetic abnormality and provide important genetic advice. PMID- 20018512 TI - High contrast hollow-cone dark field transmission electron microscopy for nanocrystalline grain size quantification. AB - In this paper, we describe hollow-cone dark field (HCDF) transmission electron microscopy (TEM) imaging, with a slightly convergent beam, as an improved technique that is suitable to form high contrast micrographs for nanocrystalline grain size quantification. We also examine the various factors that influence the HCDF TEM image quality, including the conditions of microscopy (alignment, focus and objective aperture size), the properties of the materials imaged (e.g., atomic number, strain, defects), and the characteristics of the TEM sample itself (e.g., thickness, ion milling artifacts). Sample preparation was found to be critical and an initial thinning by wet etching of the substrate (for thin film samples) or tripod polishing (for bulk samples), followed by low-angle ion milling was found to be the preferred approach for preparing high-quality electron transparent samples for HCDF imaging. PMID- 20018513 TI - Inter-laboratory comparison measurements of radiochemical laboratories in Slovakia. AB - The first inter-laboratory comparison organized by the radiochemistry laboratory of Water Research Institute (WRI) in Bratislava was carried out in 1993 and since then is it realized on an annual basis and about 10 radiochemical laboratories from all over Slovakia are participating. The gross alpha and gross beta activities, and the activity concentrations of (222)Rn, tritium, and (226)Ra, and U(nat) concentration in synthetic water samples are compared. The distributed samples are covering the concentration range prevailing in potable and surface waters and are prepared by dilution of certified reference materials. Over the course of the years 1993-2008, we observed the improvement in the quality of results for most of the laboratories. However, the success rate of the gross alpha determination activity is not improving as much as the other parameters. PMID- 20018514 TI - Validation of a new TDCR system at NPL. AB - A new triple to double coincidence ratio (TDCR) system has been established at NPL. The system incorporates a spherical optical chamber, a manual sample changing facility and an integral light-tight housing. A 6 in diameter NaI(Tl) detector has been incorporated to allow 4pibeta-gamma coincidence measurements to be performed in parallel. Details of the detectors, the electronics and the overall TDCR system are given. Validation measurements using suitable low-energy beta and electron capture nuclides, i.e. (3)H and (55)Fe have been performed. The highest efficiency achieved with a (3)H solution in Ultima Gold AB in a glass vial was 53% and in a high-efficiency LS cocktail 65%. This indicates that the optical chamber is performing well. After varying the efficiency by de-focusing the PM tubes, the activity of the sources was determined. The activity concentration determined with TDCR agreed with certified values within the range of uncertainties. Further results from validation measurements and the corresponding uncertainty budgets are presented. PMID- 20018515 TI - Assessment of the suitability of different random number generators for Monte Carlo simulations in gamma-ray spectrometry. AB - The Monte Carlo method has become a valuable numerical laboratory framework in which to simulate complex physical systems. It is based on the generation of pseudo-random number sequences by numerical algorithms called random generators. In this work we assessed the suitability of different well-known random number generators for the simulation of gamma-ray spectrometry systems during efficiency calibrations. The assessment was carried out in two stages. The generators considered (Delphi's linear congruential, mersenne twister, XorShift, multiplier with carry, universal virtual array, and non-periodic logistic map based generator) were first evaluated with different statistical empirical tests, including moments, correlations, uniformity, independence of terms and the DIEHARD battery of tests. In a second step, an application-specific test was conducted by implementing the generators in our Monte Carlo program DETEFF and comparing the results obtained with them. The calculations were performed with two different CPUs, for a typical HpGe detector and a water sample in Marinelli geometry, with gamma-rays between 59 and 1800 keV. For the Non-periodic Logistic Map based generator, dependence of the most significant bits was evident. This explains the bias, in excess of 5%, of the efficiency values obtained with this generator. The results of the application-specific assessment and the statistical performance of the other algorithms studied indicate their suitability for the Monte Carlo simulation of gamma-ray spectrometry systems for efficiency calculations. PMID- 20018516 TI - Study on anti-reflection of CR-39 by the nuclear track technique. AB - Nuclear tracks can be etched into pores in which the refractive indexes gradually increase, so that the sample surface reflects very few lights back. CR-39 samples were bombarded with sulfur ions from the Tandem Accelerator HI-13 of CIAE. Three kinds of procedures were performed in the following treatments. The reflectivity was measured for all samples, untreated and treated, within the wavelengths of visible light. The results indicated that the reflectivity of the CR-39 samples modified with the track technique and multilayer coating can be as low as 0.1%. PMID- 20018517 TI - Half-life measurements of long-lived radionuclides--new data analysis and systematic effects. AB - Half-lives of long-lived radionuclides, for example, (85)Kr, (90)Sr, (108m)Ag, (133)Ba, (137)Cs, (152)Eu and (154)Eu, were measured at the Physikalisch Technische Bundesanstalt (PTB) by ionization chamber measuring systems, for some nuclides, during a period of about 30 years. In particular, the relative residuals of fitted raw data from current measurements by a modified Townsend balance (compensation method) showed small periodic yearly fluctuations. The interpretation of these fluctuations is discussed. To solve problems of instrument instabilities (fluctuations), a new method of data analysis is applied using least-squares fits of combinations of the current ratios of the various radionuclide sources with corresponding data points measured each at nearly the same time. The data are corrected for instabilities similarly to when using current ratios with a (226)Ra reference source. The new method of data analysis excludes possible systematic effects due to disequilibrium of the (226)Ra decay chain. The limits of the method are discussed including the elimination of outliers due to systematic errors. An uncertainty budget is presented with the corresponding uncertainty components. Relative residuals of the fits in the best cases of the order of +/-5x10(-4) have been achieved. The results comprise data until the end of 2008. They agree within the limits of their relative uncertainties, typically better than 1.5x10(-3), with those published earlier by PTB authors. PMID- 20018518 TI - Participation in the CCRI(II)-K2.H-3 comparison and study of the new TDCR-LS counter with 6 CPMs. AB - The "conventional" TDCR system was used for standardization of the tritiated water in the frame of the international comparison organized by CCRI(II). The prototype of the new TDCR realized at LNHB with three CPMs (Ivan et al., 2008) was improved by constructing at IFIN-HH, a system containing six channel photomultiplier tubes arranged symmetrically in a new optical chamber. The system was tested by measuring various pure beta emitters and for comparative measurements of tritiated water for validation studies. PMID- 20018519 TI - Activity measurements of 3H using the TDCR method and observation of source stability. AB - Activity measurements of (3)H were performed using the triple to double coincidence ratio method for the 2009 international comparison organized by the Bureau International des Poids et Mesures (BIPM). The activity obtained by using the FPGA (field programmable gate array) acquisition system and software event analysis was compared to that using MAC3 (Module d'Acquisition de Coincidences triples). Agreement to better than 0.4% was obtained. The stability of sources prepared in Ultima Gold, Ultima Gold LLT, Insta-Gel Plus and Optiphase "Hisafe"-3 over 140 days is presented. PMID- 20018520 TI - Multimodality imaging in the management of aorto-coronary saphenous vein graft aneurysm. PMID- 20018521 TI - The modified Ross operation using a Dacron prosthetic vascular jacket does prevent pulmonary autograft dilatation at 4.5-year follow-up. AB - OBJECTIVE: Following the Ross operation, pulmonary autografts tend to dilate over time. This study researches the fate of the pulmonary autograft - at 4.5 years following the modified Ross operation - with special reference to the impact of the modification on (a) pulmonary autograft dilatation, (b) the neo-aortic root geometry, (c) neo-aortic valve function and (d) the coronary artery reserve. METHODS: A total of 26 patients who underwent the Ross operation were included in this study; of these, 13 consecutive patients underwent a modified Ross operation in which the free-standing autograft root was supported externally by a Dacron vascular prosthetic jacket (DVPJ). These patients were compared to a cohort of 13 matched patients who were operated on using the conventional Ross technique; all patients were followed up prospectively by echocardiography studies. The patients who underwent the modified Ross operation were also subjected to bicycle ergometry. RESULTS: At the 47-month median follow-up, there was no significant increase in the size of the entire neo-aortic root in the patients who underwent the modified Ross operation; in addition, the geometry of the neo-aortic root was also preserved and the left ventricular function had improved significantly, whilst the aortic valve function and excursion remained satisfactory. All patients, with one exception, in the modified Ross operation group exhibited normal exercise capacity. By contrast, there were significant differences in diameters of the aortic root - between the two surgical techniques in favour of the modified Ross technique - following a median follow-up of 23 months in the patients subjected to the conventional Ross operation. CONCLUSIONS: Provision of external support to the entire pulmonary autograft with a DVPJ prevents its dilatation following free-standing pulmonary autograft Ross operation when evaluated at the 4.5-year follow-up. The function and the geometry of the neo aortic root are not affected negatively by this modification and the patients demonstrated normal exercise capacity. PMID- 20018522 TI - Re: dissected Biovalsalva prosthesis. PMID- 20018523 TI - Off-pump coronary artery bypass surgery versus standard linear or pulsatile cardiopulmonary bypass: endothelial activation and inflammatory response. AB - OBJECTIVE: Poor outcomes after coronary artery bypass grafting (CABG) have been linked to perioperative endothelial activation and systemic inflammatory responses. The use of pulsatile cardiopulmonary bypass (PCPB) or off-pump CABG (OPCABG) may minimise these phenomena. We compared biochemical and clinical outcomes among patients who underwent CABG with PCPB, CABG with linear CPB (LCPB) or OPCABG. METHODS: Sixty consecutive patients undergoing isolated elective CABG were prospectively randomised trial to receive pulsatile CPB (group A, 20 patients), linear CPB (group B, 20 patients) or OPCABG (group C, 20 patients). Levels of proinflammatory cytokines (interleukins-2, -6, and -8), anti inflammatory cytokines (interleukin-10) and endothelial markers (vascular endothelial growth factor (VEGF), monocyte chemo-attractant protein (MCP)-1) were measured before, during and after surgery. RESULTS: VEGF and MCP-1 levels increased significantly during surgery in all groups, but they increased the least and were the lowest overall with OPCABG. They rose most and peaked overall with LCPB. Interleukin-2 levels remained stable during OPCABG but decreased equally during PCPB and LCPB. Interleukin-6 and -8 levels rose significantly during both types of CPB versus OPCABG. Interleukin-10 levels increased significantly in all groups during surgery, but they rose least and were the lowest overall with OPCABG and rose most and were the highest overall with PCPB. Intubation times, intensive care unit (ICU) stay and hospital stay were significantly longer in the LCPB group than the other two groups. CONCLUSIONS: LCPB appears to promote endothelial activation and cytokine secretion, which may delay recovery. OPCABG was associated with slight endothelial activation and cytokine response. PCPB significantly attenuates endothelial/cytokine leakage, resulting in hospital outcomes comparable with those after OPCABG. PMID- 20018526 TI - Information integration based predictions about the conscious states of a spiking neural network. AB - This paper describes how Tononi's information integration theory of consciousness was used to make detailed predictions about the distribution of phenomenal states in a spiking neural network. This network had approximately 18,000 neurons and 700,000 connections and it used models of emotion and imagination to control the eye movements of a virtual robot and avoid 'negative' stimuli. The first stage in the analysis was the development of a formal definition of Tononi's theory of consciousness. The network was then analysed for information integration and detailed predictions were made about the distribution of consciousness for each time step of recorded activity. This work demonstrates how an artificial system can be analysed for consciousness using a particular theory and in the future this approach could be used to make predictions about the phenomenal states associated with biological systems. PMID- 20018527 TI - Aspergillus pancarditis manifesting as hospital-acquired infection: report of two cases and review of literature. AB - Nosocomial cardiac infections are most often related to interventions performed within the preceding one to two months of hospital admission and usually affect the endocardium or prosthetic devices. These can be sometimes caused by fungi, especially the molds. This is a report of rare fungal pancarditis seen as hospital-acquired infection in two patients who were admitted in the intensive care unit for leptospirosis. The cardiac manifestation was part of systemic mycosis; mechanical ventilation, administration of steroids and hepatorenal failure were the risk factors. Incidentally, both the patients had underlying intrinsic cardiac disease. PMID- 20018525 TI - Insulin-like actions of glucagon-like peptide-1: a dual receptor hypothesis. AB - GLP-1 (9-36)amide is the cleavage product of GLP-1(7-36) amide, formed by the action of diaminopeptidyl peptidase-4 (Dpp4), and is the major circulating form in plasma. Whereas GLP-1(7-36)amide stimulates glucose-dependent insulin secretion, GLP-1(9-36)amide has only weak partial insulinotropic agonist activities on the GLP-1 receptor, but suppresses hepatic glucose production, exerts antioxidant cardioprotective actions and reduces oxidative stress in vasculature tissues. These insulin-like activities suggest a role for GLP-1 (9 36)amide in the modulation of mitochondrial functions by mechanisms independent of the GLP-1 receptor. In this paper, we discuss the current literature suggesting that GLP-1(9-36)amide is an active peptide with important insulin-like actions. These findings have implications in nutrient assimilation, energy homeostasis, obesity, and the use of Dpp4 inhibitors for the treatment of diabetes. PMID- 20018528 TI - Cancer informatics for the clinician: an interaction database for chemotherapy regimens and antiepileptic drugs. AB - BACKGROUND: Cancer patients are at high risks of manifesting drug-drug interactions (DDIs) which can potentiate serious negative outcomes. It is important for clinicians to make accurate, timely, safe and effective decisions with regards to drug use in the patient. OBJECTIVE: To provide clinicians with an oncology-specific drug interaction database that is relevant to their daily practice. This database focuses on DDIs with anticancer drugs (ACDs), single- and multiple-agent chemotherapy regimens. METHODS: Drug-related and interaction related information between anticancer and antiepileptic drugs were compiled from drug information handbooks and databases, package inserts, and published literature from PubMed. Web documents were served from a modified web server with a database engine and programming interface constructed using Adobe software and various programming scripts. RESULTS: OncoRx is an oncology-specific database whose structure is designed around chemotherapy regimens and generic drug names used in anticancer treatment. OncoRx currently comprises of 117 ACDs and 256 single-agent and combination chemotherapy regimens. It covers all the major classes of ACDs and their frequently prescribed chemotherapy regimens, and can detect up to over 2000 interactions with 24 antiepileptic drugs. Data provided to clinicians include pharmacokinetic parameters of the drugs, information regarding the chemotherapy regimens and the detected DDIs. CONCLUSION: OncoRx is able to identify DDIs between ACDs and adjuvant drug therapy. This is the first database of its kind to be able to detect interactions for combination chemotherapy regimens. This tool will assist clinicians in improving clinical response and reducing adverse effects based on the therapeutic and toxicity profiles of the drugs. PMID- 20018529 TI - Imaging of patients with hippocampal sclerosis at 7 Tesla: initial results. AB - RATIONALE AND OBJECTIVES: Focal epilepsies potentially can be cured by neurosurgery; other treatment options usually remain symptomatic. High-resolution magnetic resonance (MR) imaging is the central imaging strategy in the evaluation of focal epilepsy. The most common substrate of temporal epilepsies is hippocampal sclerosis (HS), which cannot always be sufficiently characterized with current MR field strengths. Therefore, the purpose of our study was to demonstrate the feasibility of high-resolution MR imaging at 7 Tesla in patients with focal epilepsy resulting from a HS and to improve image resolution at 7 Tesla in patients with HS. MATERIALS AND METHODS: Six patients with known HS were investigated with T1-, T2-, T2(*)-, and fluid-attenuated inversion recovery weighted sequences at 7 Tesla with an eight-channel transmit-receive head coil. Total imaging time did not exceed 90 minutes per patient. RESULTS: High resolution imaging at 7 Tesla is feasible and reveals high resolution of intrahippocampal structures in vivo. HS was confirmed in all patients. The maximum non-interpolated in-plane resolution reached 0.2 x 0.2 mm(2) in T2(*) weighted images. The increased susceptibility effects at 7 Tesla revealed identification of intrahippocampal structures in more detail than at 1.5 Tesla, but otherwise led to stronger artifacts. Imaging revealed regional differences in hippocampal atrophy between patients. The scan volume was limited because of specific absorption rate restrictions, scanning time was reasonable. CONCLUSIONS: High-resolution imaging at 7 Tesla is promising in presurgical epilepsy imaging. "New" contrasts may further improve detection of even very small intrahippocampal structural changes. Therefore, further investigations will be necessary to demonstrate the potential benefit for presurgical selection of patients with various lesion patterns in mesial temporal epilepsies resulting from a unilateral HS. PMID- 20018530 TI - Ultrasound imaging of the sural nerve: ultrasound anatomy and rationale for investigation. AB - BACKGROUND: Damage to the sural nerve (SuN) may arise from surgical stripping or thermal ablation of the small saphenous vein (SSV). OBJECTIVE: This study aims to demonstrate that visualisation of the SuN and its point of contact with the SSV ('risk point') using ultrasound imaging can be achieved in routine clinical practice. TYPE OF STUDY: This is a cohort study. PATIENTS: Fifteen normal subjects and five patients with chronic venous insufficiency (CVI) (two with a dilated, incompetent SSV). METHOD: The SuN was identified using high-resolution ultrasound imaging using 14- and 18-MHz probes. Two manoeuvres were found to improve visualisation: (1) the contrast of the nerve was increased compared with the other tissues by varying the angle of insonation; and (2) the transducer was moved up and down the limb for a short distance during transverse imaging of the calf. The muscles and other soft tissues appeared 'out of focus', whereas the SuN retained both shape and echogenicity. Once the nerve has been identified, proceeding proximally, the point of separation of the two components is often detectable. It is then possible to follow the two different nerves observing the medial sural cutaneous nerve (MSCN) inside the 'triangle' of connective tissue below the SSV joining the tibial nerve and the lateral sural cutaneous nerve (LCSN) joining the common peroneal nerve, which runs inside a tiny fascial duplication. The extent of nerves, which were identified, was recorded in each limb as well as their anatomical distribution. RESULTS: The SuN and the point at which it might be at risk were identified on ultrasound images in 39 of 40 limbs (97%) studied. In transverse section, it was readily identified within the saphenous compartment. It lies in close proximity to the SSV only in the distal third of the limb, where the two components of the nerve: MSCN, a branch of the tibial nerve; and LSCN, a branch of the common peroneal nerve join together. The relationship between the SuN and the SSV is very variable, with the nerve running separately or in close contact with the vein for variable distances, in many different combinations. CONCLUSIONS: The SuN and 'risk point' can be identified by ultrasonography (US). We propose that this technique could be used to prevent damage to the SuN during surgical or thermal ablation of the SSV and during Achilles tendon surgery. PMID- 20018531 TI - Combined simultaneous basilic and brachial vein transposition. A new technique to create an autologous vascular access. AB - In this report, we present a technique to extend the function of an antecubital arteriovenous fistula in which both the basilic and the brachial veins were simultaneously transposed to create an autologous graft in the arm. This procedure may particularly be applicable for patients in whom, although a brachio cephalic fistula, anastomosing the brachial artery and the perforating antecubital vein, has been previously performed and has remained patent arterialising the deep arm veins, the cephalic vein has failed to mature or has been thrombosed after multiple punctures. Our preliminary experience in eight patients has shown satisfactory outcome. PMID- 20018532 TI - Prenatal diagnosis of hemophilia A, presenting a new mutation in the FVIII gene. PMID- 20018533 TI - Measurement of non-restorative sleep in insomnia: A review of the literature. AB - OBJECTIVE: Non-restorative sleep (NRS) is a core symptom of insomnia, typically defined as a subjective feeling of being unrefreshed upon awakening. NRS symptoms have been less consistently studied than other symptoms of insomnia, and there is no consensus regarding measurement of NRS. Given its subjective nature, patient reported outcome (PRO) instruments are important for evaluating NRS symptom severity and treatment-related changes. The objective of this literature review was to identify PROs used in insomnia populations that purport to measure NRS, and to evaluate their psychometric characteristics. METHODS: A comprehensive review of PRO instruments used in insomnia was conducted; instruments were reviewed for NRS content by two independent reviewers; a total of 26 instruments identified to contain NRS content were retained for further review. RESULTS: Of the 26 instruments reviewed, 23 contained at least one item evaluating subjective nighttime aspects of NRS; 17 contained at least one item evaluating daytime aspects of NRS. Only the Sleep Assessment Questionnaire contained a specific NRS domain score. However, little published evidence was available regarding measurement properties of the NRS domain in insomnia populations. CONCLUSIONS: There is currently no reliable and well-validated PRO instrument available for specifically evaluating NRS symptom severity and response to interventions in insomnia populations. Reliable and valid measurement tools are needed to measure the symptom of NRS in insomnia. PMID- 20018534 TI - Evaluation of candidate genes as a cause of chondrodysplasia in Labrador retrievers. AB - Chondrodysplasia (CD) is a disabling, hereditary disease in Labradors with short limbs, warranting genetic screening in families at risk. Segregation analysis of eight litters with 13 affected dogs showed that autosomal recessive inheritance was consistent with the observed incidence of CD in the litters. Possible involvement of eight candidate collagen genes (COL9A1, COL9A2, COL9A3, COMP, MATN3, COL2A1, COL11A1 and COL11A2) and of a sulfate transporter glycoprotein (SLC26A2) gene in eight affected dogs and in 14 related control Labradors was investigated. Assuming recessive inheritance, the candidate genes could not be implicated in CD. PMID- 20018535 TI - Efficacy and safety of a single intrathecal methylprednisolone bolus in chronic complex regional pain syndrome. AB - Activated immune cells in the spinal cord may play an important role in the development and maintenance of neuropathic pain, such as occurs in response to peripheral inflammation or tissue injury. Immune activation may therefore serve as a therapeutic target for immune modulating drugs like corticosteroids. This double-blind randomized placebo-controlled parallel-group trial aimed to investigate the efficacy and safety of a single intrathecal administration of 60 mg methylprednisolone (ITM) in chronic patients with complex regional pain syndrome (CRPS). The primary outcome measure was change in pain (pain intensity numeric rating scale; range 0-10) after 6 weeks. With 21 subjects per group the study had a 90% power to detect a clinically relevant difference (> or = 2 points). After 21 patients (10 on ITM) were included, the trial was stopped prematurely after the interim analysis had shown that ITM had no effect on pain (difference in mean pain intensity numeric rating scale at 6 weeks 0.3, 95% confidence interval -0.7 to 1.3) or any other outcome measure. We did not find any difference in treatment-emergent adverse events between the ITM and placebo group. We conclude that a single bolus administration of ITM is not efficacious in chronic CRPS patients, which may indicate that spinal immune activation does not play an important role in this phase of the syndrome. PMID- 20018536 TI - Increased pain sensitivity in alcohol withdrawal syndrome. AB - Withdrawal from analgesic and addictive substances such as opioids or ethanol is associated with increased sensitivity to sensory stimulation in animal models. Here, we investigated perception of innocuous and noxious thermal or electric stimuli applied to the left hand or sternum in 30 male patients undergoing withdrawal from alcohol, 30 male abstained alcoholics and matched controls. The alcohol withdrawal scale and the Banger score were obtained to estimate the severity of withdrawal. In addition, the Beck depression inventory was used to estimate the influence of depressive symptoms on pain perception. The data presented provide substantial evidence that subjects undergoing alcohol withdrawal show increased heat pain sensitivity. Interestingly, this effect was observed both on the left hand and sternum. Pain thresholds and tolerances of electric stimuli did not differ between groups. However, in a subgroup analysis, a higher sensitivity for electrical pain thresholds and tolerances was observed in those patients that were identified to require pharmacological treatment for withdrawal according to disease severity. Furthermore, the perceived painful thermal and electrical sensation was substantially influenced by the affective state of patients. No differences were found between patients of the abstained group and control subjects for any pain parameter. In conclusion, we demonstrate withdrawal-induced hyperalgesia upon thermal stimulation in patients. Since the influence of affective symptoms on pain perception during withdrawal is remarkable, we assume that peripheral and central mechanisms might account for this finding, which should be assessed in detail in future studies. PMID- 20018537 TI - Effects of pain management on sleep in preterm infants. AB - BACKGROUND: This study was conducted to gain better understanding of the prolonged effects of pain and pain management on preterm infants' sleep. AIM: The hypothesis was that the sleep structure in very preterm infants is different after painful procedures with pain management (facilitated tucking by parents (FTP), oral glucose, and oxycodone) than without pain management (oral water as placebo). METHODS: A prospective randomized placebo-controlled cross-over trial design was used. Thirteen-hour polysomnographic recordings were conducted when the study infants (n=18) were at a post-conceptional age of 28-32 weeks. During the recordings, the standardized nursing care periods were carried out with different forms of pain management administered at 3-h intervals. Sleep structure was analyzed before and after the interventions. The main hypothesis was analyzed using mixed models. RESULTS: During the first post-intervention hour, the amount of rapid eye movement (REM) sleep decreased after all interventions regardless of pain management (p<0.001). However, the oxycodone treatment further reduced the amount of REM sleep to 48.0% (SD 14.9) compared to other interventions: oral glucose to 64.4% (SD 12.8), (p<0.001); placebo to 62.9% (SD 16.1), (p<0.001); and FTP to 61.6% (SD 1.9), (p=0.004). In addition, sleep onset comprised non-rapid eye movement (NREM) sleep more frequently after oxycodone (50%) compared to placebo (6%, p=0.006), oral glucose (11%, p=0.019) or FTP (17%, p=0.056). CONCLUSION: Pain management with oxycodone markedly altered the structure of the subsequent sleep period. This reduced amount of REM sleep may have consequences for brain development in preterm infants. PMID- 20018538 TI - Measurement and interpretation of 15N-1H residual dipolar couplings in larger proteins. AB - A decade ago, Dr. L.E. Kay and co-workers described an ingenious HNCO-based triple-resonance experiment from which several protein backbone RDCs can be measured simultaneously (Yang et al. (1999) [1]). They implemented a J-scaling technique in the (15)N dimension of the 3D experiment to obtain the NH RDCs. We have used this idea to carry out J-scaling in a 2D (15)N-(1)H-TROSY experiment and have found it to be an excellent method to obtain NH RDCs for larger proteins upto 70 kDa, far superior to commonly used HSQC in-phase/anti-phase and HSQC/TROSY comparisons. Here, this method, dubbed "RDC-TROSY" is discussed in detail and the limits of its utility are assessed by simulations. Prominent in the latter analysis is the evaluation of the effect of amide proton flips on the "RDC-TROSY" linewidths. The details of the technical and computational implementations of these methods for the determination of domain orientations in 45-60 kDa Hsp70 chaperone protein constructs are described. PMID- 20018539 TI - [Satisfaction among patients attending outpatient preventive medicine clinic]. AB - INTRODUCTION: To evaluate satisfaction among patients attending the outpatient clinics of a Preventive Medicine Department and to identify any problems with the care they receive. MATERIAL AND METHODS: We performed a cross-sectional study in outpatient clinics of the Preventive Medicine Department, at Hospital General Universitario in Alicante, during January 2009. A questionnaire was given to all patients after their visit. Patients were satisfied if the final score was > or = 8. Socio-demographic variables were: age, sex, educational level, marital status and type of user (external or internal). chi(2) were used to study the association between these variables and satisfaction. RESULTS: The response rate was 84.4% (119/141), 94.1% were correctly completed (112/119), and 82.1% had a final score of > or = 8. The worst aspects were: comfort of waiting room, signs and wait time (percentage dissatisfied: 54.5%, 46.4% and 30.4%, respectively). The rest of the items obtained the following satisfaction percentages: medical treatment (97.3%), privacy of care (95.5%), information received (93.7%), easy next appointment (93.7%), visit length (92.9%), clarity of information (92.9%), to arrange the first appointment (90.2%), nursing treatment (87.5%), time to date of appointment (81.2%). Educational level was the only variable significantly associated with satisfaction. CONCLUSIONS: General satisfaction was good, although aspects like waiting time and signs could be better, if changes were made to management of appointments and the signs in the Hospital Department were increased. PMID- 20018540 TI - [Validity of synthetic indicators for measuring performance]. PMID- 20018541 TI - [Leadership in patient safety]. PMID- 20018542 TI - [Patient and caregiver satisfaction with rehabilitation services after stroke]. AB - OBJECTIVES: To assess patient and caregiver satisfaction with the regular rehabilitation care after stroke. To identify patient features or aspects of the rehabilitation programme related to lower rates of satisfaction in our area. MATERIAL AND METHODS: Cross-sectional study of 74 stroke in-patients admitted from June 2005 to June 2006 (70.3% men, age 65.4 + or - 12.2 years, Barthel discharge 70.5 + or - 27). A telephone Satisfaction questionnaire (Pound 1999) was administered at 13.6 + or - 3.1 months to assess satisfaction regarding in patient care, therapy and recovery, and services after hospital discharge. Three items addressed to caregiver satisfaction were also included. A total of 84 caregivers were identified: 71% relatives (partner or children), of which 80.9% were women. RESULTS: The satisfaction questionnaire was completed by 52.9% of patients. Over 80% of interviewed patients were completely satisfied with in patient care, and 52.7% were satisfied with therapy and recovery during admission, but more patients were dissatisfied with the amount of therapy received. The lowest rate of satisfaction was related to the services after discharge, with less than 25% of patients satisfied. A total of 83.1% of caregivers completed the questionnaire, of which 74.1% were satisfied with the information provided. Stroke educational training was given to 63.5% of caregivers, and 42.4% were given support after discharge. Only the presence of depression during admission and time passed after stroke were significantly related to patient satisfaction. (p<0.05). CONCLUSIONS: Dissatisfaction with some aspects of stroke rehabilitation is considerable in our care area. Despite general patient and caregiver satisfaction regarding information provided, high dissatisfaction rates are also detected related to the amount of therapy and the social support provided after hospital discharge. According to these results, systematic information and training programs for patients and caregivers should be included in stroke rehabilitation, as well as providing social support at discharge. PMID- 20018543 TI - [Quality of the Intensive Medicine admission criteria by perceived by hospital doctors]. PMID- 20018544 TI - Narghile (hookah, shisha) tobacco smoking: expect the unexpected everywhere and not only in Syria. PMID- 20018545 TI - Serologic response to rabies pre-exposure vaccination in persons with potential occupational exposure in Singapore. AB - Sixty-six animal workers received primary rabies vaccination with purified Vero cell vaccine (PVRV, Verorab). One year later, 26 (39%) demonstrated antibody titers below the recommended minimum of 0.5 IU/ml, and required a booster. All 15 of a separate group reporting primary vaccination with at least one booster had titers above 0.5 IU/ml 1 year later, demonstrating long-term boostable immunity. Rabies antibody titers should be checked 1 year after primary rabies vaccination in persons at high risk of frequent rabies exposure. If access to serological surveillance is unavailable, such high-risk individuals should receive booster vaccination. PMID- 20018546 TI - A hypothetical explanatory model for meningococcal meningitis in the African meningitis belt. AB - Despite much progress in surveillance and biological research, no explanation exists to date for the epidemic pattern of meningitis in the African meningitis belt, which is required to mathematically model the impact of vaccine strategies or to predict epidemics. This paper presents a hypothetical explanatory model for epidemic meningococcal meningitis. Four incidence patterns are defined as model states, including endemic incidence during the rainy season, ubiquitous hyperendemicity during the dry season, occasional localized epidemics, and-at the regional level-regular epidemic waves spanning over communities or years. While the transition from endemic to hyperendemic situation in a community is caused by an increase in risk of meningitis given colonization by a virulent meningococcus (due to damage of the pharyngeal mucosa by dry climate), the transition from hyperendemic to epidemic situation involves increased pharyngeal colonization and transmission (possibly caused by viral respiratory infection epidemics). The described mechanisms are sufficient to explain the 10- to 100-fold incidence increase that both transitions usually imply. Epidemic waves occur if new meningococcal strains which escape pre-existing immunity, enter the population. Future research should include the impact of viral co-infection on bacterial colonization and invasion. PMID- 20018547 TI - [Erythropoietin and radiotherapy]. AB - Erythropoietin (EPO) is a glycoprotein hormone. This hormone is a growth factor for red blood cells precursors in the bone marrow. The decrease of oxygen partial pressure, a reduced number of erythrocytes caused by bleeding or excessive destruction, or increased tissues oxygen requirements lead to increased secretion of EPO. Its action takes place on bone marrow erythroblastic cells through specific receptors. EPO stimulates the proliferation of red cell precursors stem cells in the bone marrow, thus increasing their production in one to two weeks. The effectiveness of EPO at increasing haemoglobin and improving patients' quality of life has been demonstrated by several studies. However, its use in radiotherapy remains controversial. While tumour hypoxia caused by anaemia is a factor of radio resistance and thus a source of local failure, tumour expression of EPO receptors presents a significant risk for tumour progression and neo angiogenesis, which would be increased during the administration of EPO. The purpose of this article is to answer the question: is there a place for EPO in combination with radiotherapy in the management of cancer? PMID- 20018548 TI - Symmetric corticobasal degeneration (S-CBD). AB - BACKGROUND: Corticobasal degeneration (CBD) is a neurodegenerative disease characterized pathologically by neuronal loss, gliosis and tau deposition in neocortex, basal ganglia and brainstem. Typical clinical presentation is known as corticobasal syndrome (CBS) and involves the core features of progressive asymmetric rigidity and apraxia, accompanied by other signs of cortical and extrapyramidal dysfunction. Asymmetry is also emphasized on neuroimaging. OBJECTIVE: To describe a series of cases of CBD with symmetric clinical features and to compare clinical and imaging features of these symmetric CBD cases (S-CBD) to typical cases of CBS with CBD pathology. METHODS: All cases of pathologically confirmed CBD from the Mayo Clinic Rochester database were identified. Clinical records were reviewed and quantitative volumetric analysis of symmetric atrophy on head MRI using atlas based parcellation was performed. Subjects were classified as S-CBD if no differences had been observed between right- and left sided cortical or extrapyramidal signs or symptoms. S-CBD cases were compared to 10 randomly selected typical CBS cases. RESULTS: Five cases (2 female) met criteria for S-CBD. None had limb dystonia, myoclonus, apraxia or alien limb phenomena. S-CBD cases had significantly less asymmetric atrophy when compared with CBS cases (p=0.009); they were also younger at onset (median 61 versus 66 years, p<0.05) and death (67 versus 73 years, p<0.05). Family history was present in 40% of S-CBD cases. CONCLUSIONS: CBD can have a symmetric presentation, clinically and radiologically, in which typical features of CBS, such as limb apraxia, myoclonus, dystonia and alien limb phenomenon, may be absent. PMID- 20018549 TI - Association between the dose of dopaminergic medication and the behavioral disturbances in Parkinson disease. AB - OBJECTIVES: To survey the point prevalence of impulse control and repetitive behavior disorders (ICRBs) in patients with Parkinson disease (PD) and to determine the relationship between PD medication dose and the risk of ICRBs. METHODS: A multicenter cross-sectional survey was applied to consecutive patients with PD over a 3-month period. The presence of ICRBs was screened using a modified version of the Minnesota Impulsive Disorders Interview that comprised five ICRB modules: compulsive buying, gambling, sexual behavior, eating, and punding. Data regarding the patients' clinical features and concurrent anti-PD drugs were also collected during the interview. Adjusted odds ratios (ORs) of the daily doses of dopamine agonist and L-dopa for the development of an ICRB were calculated after adjustment for clinical variables. RESULTS: Among the 1167 patients recruited, 118 (10.1%) exhibited ICRBs. Punding was the most common ICRB (4.2%), followed by compulsive eating (3.4%), sexual behaviors (2.8%), buying (2.5%), and gambling (1.3%). Two or more ICRBs were present concomitantly in 34 of these 118 patients (28.8%). There were dose-response relationships between the dopamine agonist dose and the ORs for compulsive buying, gambling and sexual behaviors. On the other hand, the OR for punding was positively correlated with the dose of L-dopa. The OR for compulsive eating was not associated with the dose of dopamine agonist or L-dopa. CONCLUSIONS: The dose of dopaminergic medication is significantly associated with the development of ICRB, except compulsive eating, in PD. PMID- 20018550 TI - Who is affected more by air pollution-sick or healthy? Some evidence from a health survey of schoolchildren living in the vicinity of a coal-fired power plant in Northern Israel. AB - OBJECTIVE: To evaluate the effects of exposure to air pollution by NO(x) and SO(2) on the development of pulmonary function of children, characterized by different health status. METHODS: A cohort of 1181 schoolchildren from the 2nd to 5th grades, residing near a major coal-fired power plant in the Hadera district of Israel, were subdivided into three health status groups, according to the diagnosis given by a physician at the beginning of the study period in 1996: (a) healthy children; (b) children experiencing chest symptoms, and (c) children with asthma or spastic bronchitis. Pulmonary Function Tests (PFTs) were performed twice (in 1996 and 1999) and analyzed in conjunction with air pollution estimates at the children's places of residence and several potential confounders-height, age, gender, parental education, passive smoking, housing density, length of residence in the study area and proximity to the main road. RESULTS: A significant negative association was found between changes in PFT results and individual exposure estimates to air pollution, controlled for socio-demographic characteristics of children and their living conditions. A sensitivity analysis revealed a decrease in the Forced Expiratory Volume during the First Second (FEV(1)) of about 19.6% for children with chest symptoms, 11.8% for healthy children, and approximately 7.9% for children diagnosed with asthma. Results of a sensitivity test for the Forced Vital Capacity (FVC) were found to be similar. CONCLUSION: Exposure to air pollution appeared to have had the greatest effect on children with chest symptoms. This phenomenon may be explained by the fact that this untreated symptomatic group might experience the most severe insult on their respiratory system as a result of exposure to ambient air pollution, which is reflected by a considerable reduction in their FEV(1) and FVC. Since asthmatic children have lower baseline and slower growth rates, their PFT change may be affected less by exposure to air pollution, reflecting a well known relationship between pulmonary function change and height growth, according to which age specific height is very similar for preadolescent children, but shifts upward with age during the growth spurt. PMID- 20018553 TI - Metal complexes in medicine with a focus on enzyme inhibition. AB - Since the clinical success of cisplatin and its derivatives, considerable effort has been expended by academics and pharmacological companies to the development of novel metal-based drugs. DNA is believed to be the main target of cisplatin, and there have been extensive studies on the binding between metal complexes and DNA targets. Recently, new light has been shed on the discovery of metal-based drugs that inhibit enzymatic activities or even target proteins directly. This review highlights some exciting results published recently on the development of platinum, gold, and ruthenium complexes as enzyme inhibitors for potential therapeutic applications. PMID- 20018551 TI - Transforming growth factor beta (TGF-beta) and inflammation in cancer. AB - The transforming growth factor beta (TGF-beta) has been studied with regard to the regulation of cell behavior for over three decades. A large body of research has been devoted to the regulation of epithelial cell and derivative carcinoma cell populations in vitro and in vivo. TGF-beta has been shown to inhibit epithelial cell cycle progression and promote apoptosis that together significantly contribute to the tumor suppressive role for TGF-beta during carcinoma initiation and progression. TGF-beta is also able to promote an epithelial to mesenchymal transition that has been associated with increased tumor cell motility, invasion and metastasis. However, it has now been shown that loss of carcinoma cell responsiveness to TGF-beta stimulation can also promote metastasis. Interestingly, enhanced metastasis in the absence of a carcinoma cell response to TGF-beta stimulation has been shown to involve increased chemokine production resulting in recruitment of pro-metastatic myeloid derived suppressor cell (MDSC) populations to the tumor microenvironment at the leading invasive edge. When present, MDSCs enhance angiogenesis, promote immune tolerance and provide matrix degrading enzymes that promote tumor progression and metastasis. Further, the recruitment of MDSC populations in this context likely enhances the classic role for TGF-beta in immune suppression since the MDSCs are an abundant source of TGF-beta production. Importantly, it is now clear that carcinoma-immune cell cross-talk initiated by TGF-beta signaling within the carcinoma cell is a significant determinant worth consideration when designing therapeutic strategies to manage tumor progression and metastasis. PMID- 20018552 TI - Dangerous liaisons: STAT3 and NF-kappaB collaboration and crosstalk in cancer. AB - Transcriptional factors of the NF-kappaB family and STAT3 are ubiquitously expressed and control numerous physiological processes including development, differentiation, immunity, metabolism and cancer. Both NF-kappaB and STAT3 are rapidly activated in response to various stimuli including stresses and cytokines, although they are regulated by entirely different signaling mechanisms. Once activated, NF-kappaB and STAT3 control the expression of anti apoptotic, pro-proliferative and immune response genes. Some of these genes overlap and require transcriptional cooperation between the two factors. The activation of and interaction between STAT3 and NF-kappaB plays a key role in controlling the dialog between the malignant cell and its microenvironment, especially with inflammatory/immune cells that infiltrate tumors. Quite often, cytokines whose expression is induced in response to NF-kappaB in immune cells of the tumor microenvironment lead to STAT3 activation in both malignant and immune cells. While within malignant and pre-malignant cells STAT3 exerts important oncogenic functions, within inflammatory cells it may also suppress tumor promotion through its anti-inflammatory effects. Other interactions and forms of crosstalk between NF-kappaB and STAT3 include physical interaction between the two, cooperation of these factors at gene promoters/enhancers, the NF-kappaB dependent expression of inhibitors of STAT3 activation and the participation of STAT3 in inflammatory cells in the negative regulation NF-kappaB. Despite these versatile and occasionally antagonistic interactions, NF-kappaB and STAT3 cooperate to promote the development and progression of colon, gastric and liver cancers. In addition to explaining the molecular pathogenesis of cancer, these interactions also offer opportunities for the design of new therapeutic interventions. PMID- 20018554 TI - Spectroscopic study of 2-, 4- and 5-substituents on pKa values of imidazole heterocycles prone to intramolecular proton-electrons transfer. AB - New 2-(1H-imidazol-2-yl)phenols (L1Et-L8tBuPt) bearing a phenolic proton in the vicinity of the imidazole base were prepared and characterized. Experimental studies of the dependence of their protonation/deprotonation equilibrium on substituent identities and intramolecular hydrogen bonding tendencies were carried out using electronic absorption spectroscopy at varying pH values. In order to make comparison, 2-(anthracen-10-yl)-4,5-diphenyl-1H-imidazole (L9Anthr) bearing no phenolic proton and 4,5-diphenyl-2-(4,5-diphenyl-1H-imidazol-2-yl)-1H imidazole (L10BisIm) bearing two symmetrical imidazole base fragments were also prepared and experimentally investigated. DFT calculations were carried out to study frontier orbitals of the investigated molecules. While electron-releasing substituents produced increase in protonation-deprotonation pK(a)s for the hydroxyl group, values for the imidazole base were mainly affected by polarization of the imidazole ring aromaticity across the 2-imidazole carbon and the 4,5-imidazole carbons axis of the imidazole ring. It was concluded that electron-releasing substituents on the phenol ring and/or electron-withdrawing substituents on 4,5-imidazole carbons negatively affects donor strengths/coordination chemistries of 2-(1H-imidazol-2-yl)phenols, and vice versa. Change of substituents on the phenol ring significantly altered the donor strength of the imidazole base. The understanding of pK(a) variation on account of electronic effects of substituents in this work should aid the understanding of biochemical properties and substituent environments of imidazole-containing biomacromolecules. PMID- 20018555 TI - Electronic absorption study on acid-base equilibria for some pyrimidine derivatives containing semi- and thiosemicarbazone moiety. AB - The UV-vis spectra of recently synthesized 5-benzoyl-1 (methylphenylmethyleneamino)-4-phenyl-1H-pyrimidine-2-one, (I), and 5-benzoyl-1 (methylphenylmethyleneamino)-4-phenyl-1H-pyrimidine-2-thione, (II) were studied in aqueous methanol (5%, v/v methanol). The nature of the electronic transitions and the roles of carbonyl oxygen of I and thiocarbonyl sulfur of II on the behavior of UV-vis spectra were discussed. Acid-base equilibria of the compounds against varying pH and pK(a) values related equilibria were determined at an ionic strength of 0.10 M by using the Henderson-Haselbalch equation. The mean acidity constants for the protonated forms of the compounds were determined as pK(a1)=5.121, pK(a2)=7.929 and pK(a3)=11.130 for I and pK(a1)=4.684, pK(a2)=7.245 and pK(a3)=10.630 for II. The preferred dissociation mechanisms were discussed based on UV-vis data and a mechanism was proposed for each compound. PMID- 20018556 TI - Synthesis and spectral investigations of Mn(II) complexes of pentadentate bis(thiosemicarbazones). AB - Five Mn(II) complexes of bis(thiosemicarbazones) which are represented as [Mn(H(2)Ac4Ph)Cl(2)] (1), [Mn(Ac4Ph)H(2)O] (2), [Mn(H(2)Ac4Cy)Cl(2)].H(2)O (3), [Mn(H(2)Ac4Et)Cl(2)].3H(2)O (4) and [Mn(H(2)Ac4Et)(OAc)(2)].3H(2)O (5) have been synthesized and characterized by elemental analyses, electronic, infrared and EPR spectral techniques. In all the complexes except [Mn(Ac4Ph)H(2)O], the ligands act as pentadentate neutral molecules and coordinate to Mn(II) ion through two thione sulfur atoms, two azomethine nitrogens and the pyridine nitrogen, suggesting a heptacoordination. While in compound [Mn(Ac4Ph)H(2)O], the dianionic ligand is coordinated to the metal suggesting six coordination in this case. Magnetic studies indicate the high spin state of Mn(II). Conductivity measurements reveal their non-electrolyte nature. EPR studies indicate five g values for [Mn(Ac4Ph)H(2)O] showing zero field splitting. PMID- 20018557 TI - Spectroscopic characterization of charge-transfer complexes of morpholine with chloranilic and picric acids in organic media: crystal structure of bis(morpholinium 2,4,6-trinitrocyclohexanolate). AB - Electron donor-acceptor interaction of morpholine (morp) with chloranilic acid (cla) and picric acid (pa) as pi-acceptors was investigated spectrophotometrically and found to form stable charge-transfer (CT) complexes (n pi*) of [(Hmorp)(2)(cla)] and [(Hmorp)(pa)](2). The donor site involved in CT interaction is morpholine nitrogen. These complexes are easily synthesized from the reaction of morp with cla and pa within MeOH and CHCl(3) solvents, respectively. (1)HNMR, IR, elemental analyses, and UV-vis techniques characterize the two morpholinium charge-transfer complexes. Benesi-Hildebrand and its modification methods were applied to the determination of association constant (K), molar extinction coefficient (epsilon). The X-ray crystal structure was carried out for the interpretation the predict structure of the [(Hmorp)(pa)](2) complex. PMID- 20018558 TI - Fluorescence Resonance Energy Transfer between organic dyes adsorbed onto nano clay and Langmuir-Blodgett (LB) films. AB - In this communication we investigate two dyes N,N'-dioctadecyl thiacyanine perchlorate (NK) and octadecyl rhodamine B chloride (RhB) in Langmuir and Langmuir-Blodgett (LB) films with or with out a synthetic clay laponite. Observed changes in isotherms of RhB in absence and presence of nano-clay platelets indicate the incorporation of clay platelets onto RhB-clay hybrid films. AFM images confirm the incorporation of clay into hybrid films. FRET is observed in clay dispersion and LB films with and without clay. Efficiency of energy transfer is maximum in LB films with clay. PMID- 20018559 TI - Design and characterization of Cu(II) complexes from 2-benzoylpyridine benzhydrazone: crystallographic evidence for coordination versatility. AB - The syntheses and characterization of six copper(II) complexes of 2 benzoylpyridine benzhydrazone in the form of [Cu(BPB)(2)], [Cu(BPB)Cl].H(2)O, [Cu(BPB)Br], [Cu(2)(BPB)(2)](ClO(4))(2).4H(2)O, [Cu(BPB)N(3)].H(2)O, and [Cu(BPB)NCS].H(2)O.CH(3)OH are reported. The analytical methods used for the characterization of complexes include partial elemental analyses, IR, electronic and EPR spectra, conductivity measurements, magnetic susceptibility measurements and single crystal X-ray diffraction. From the crystal structure, it is clear that the hydrazone adopts the E conformation about the azo bond to attach to the metal through the N(py)-N(azo)-O chelating system. In the EPR spectra of complexes in DMF at 77K four hyperfine quartets in the parallel region could be resolved and a half field signal is observed at 1500 G for complex [Cu(2)(BPB)(2)](ClO(4))(2).4H(2)O in polycrystalline state at 298 K which gives evidence for its binuclear nature indicating a weak interaction between the two Cu(II) ions. PMID- 20018560 TI - Measuring human immunodeficiency virus type 1 RNA loads in dried blood spot specimens using NucliSENS EasyQ HIV-1 v2.0. AB - BACKGROUND: HIV-1 RNA plasma level is a key parameter for anti-viral treatment monitoring in HIV-1 infected individuals. Plasma stability and accurate measurement of clinical state is at risk when transporting from remote areas. Dried blood spot (DBS) testing can reduce this risk. OBJECTIVES: Determine the performance of NucliSENS EasyQ HIV-1 v2.0 for DBS. STUDY DESIGN: 100 HIV-1 negative, and 129 HIV-1 spiked blood specimens (2180 copies/ml) were used for diagnostic specificity and system robustness. Analytical performance was tested in the range 50-85,000,000 copies/ml. Clinical reactivity was measured with specimens obtained from 224 HIV-1 infected individuals. HIV-1 RNA stability was analyzed after applying several different storage conditions. RESULTS: Diagnostic specificity was 100% and system robustness was demonstrated by 100% detection rate without invalids. Limit of detection (95% detection rate) was 800 copies/ml. Linear results were obtained over the whole range tested. For clinical specimens, percentage positive results were comparable for DBS (57%) and plasma (58%). DBS quantification was on average 0.36log10 lower as compared to plasma. Specimen stability was demonstrated for 1 week at 55 degrees C/60% humidity, 3 weeks at 37 degrees C/80% humidity, 9 weeks at 37 degrees C/40% humidity, 3 months at -20 degrees C/70% humidity, 3 weeks at 4 degrees C/100% humidity, 9 months at room temperature (15-30 degrees C), and 9 weeks shipment simulation. CONCLUSION: Results obtained fully support the use of DBS for the NucliSENS EasyQ HIV-1 v2.0 assay. These findings are especially of importance in cases that plasma stability is currently at risk due to for example, long transport routes from remote areas under less controlled conditions. PMID- 20018561 TI - The life history of Flabellula baltica Smirnov (Gymnamoebae, Rhizopoda): adaptations to a spatially and temporally heterogeneous environment. AB - The polymorphic life history of the marine naked amoeba Flabellula baltica was studied. It can be interpreted in terms of adaptations to an environment that is patchy in time and space and it represents trade-off between longevity during starvation and the ability to initiate multiplication soon after food resource become available. The life history also represents bet hedging in that different cells within a clonal culture may respond in different ways when food is depleted. PMID- 20018563 TI - Comparing the effects of two educational methods of intravenous drug rate calculations on rapid and sustained learning of nursing students: formula method and dimensional analysis method. AB - Accurate calculations of flow rate of intravenous medication are crucial to safe patient care. Flow rate of these medications can be calculated using either the formula or dimensional analysis method. However, few studies have assessed the effectiveness of this teaching method or any other method of calculating drug dosages. The purpose of this study was to compare the effects of formula and dimensional analysis educational methods of intravenous drug rate calculations on nursing students' rapid and sustained learning. This was an experimental study in which 42s year nursing students participated. They were divided randomly into two groups, control and experimental and the drug calculating skills were taught to them through formula method (control group) and dimensional analysis method (experimental group), respectively. Before the education, immediately, and three months after the teaching intervention, the students' skills were examined using a drug calculation test. The results showed that no significant difference between the two groups in pre-test and post-test 1 scores (P>0.05), but did show a significant difference between the two groups in post-test 2 scores (P<0.05). The mean of the scores differences of pre-test and post-test 2 were 9.8+/-3.34 and 12.85+/-3.07 in formula method group and dimensional analysis group, respectively, which showed significant difference between two groups. This study showed that immediate post education intervention learning was significantly better in both groups, but, in the dimensional analysis method group, the sustained learning rate was significantly better than in the formula method group. PMID- 20018562 TI - Microevolution and speciation in Thalassiosira weissflogii (Bacillariophyta). AB - In this study five different molecular markers were used to: (1) infer the phylogeographic differentiation of Thalassiosira weissflogii in the Atlantic and Pacific Oceans; and (2) address the biological species status of the inferred geographic lineages. The results of the ribosomal RNA data analyses suggested that the Hawaiian isolate evolved first after which the Indonesian and the Atlantic/California strains diverged. In contrast, the tree derived from the partial sexually induced gene 1 (Sig1) data exhibited an initial divergence between the Eastern Atlantic/Western Atlantic/California and the Hawaiian/Indonesian groups after which the latter evolved into the Hawaiian and Indonesian lineages. The partial beta-tubulin phylogeny discerns an early "split" between an Eastern Atlantic/Western Atlantic clade and an Indonesia/California/Hawaii group which later differentiated into distinct Hawaiian and Indonesian/California "branches". The number of compensatory base changes (CBCs) in the ITS2 indicated that the "Atlantic group", including the California isolate, constituted a single reproductive unit and that the Indonesian and Hawaiian T. weissflogii are two different biological species with regard to each other and to the "Atlantic clade". The beta-tubulin tree contradicts the reproductive units recovered by the compensatory base change analysis due to the close affinity of the California and Indonesia strains whereas the sexually induced gene 1 phylogeny supports the existence of three biological species, despite exhibiting a temporal sequence of geographic diversification that is different from that seen in topologies derived from the ribosomal RNA data. It is hypothesized that the tree derived from the sexually induced gene 1 reflects the relative order of the evolution of reproductive isolation in the different T. weissflogii strains while the lineages in the other phylogenies depict time elapsed since common ancestry. The current investigation is the first to have shown concordance in the pattern of variation between a reasonably good biological species marker (CBCs in the ITS2) and a protein hypothesized to be involved in sperm-egg recognition/adhesion in diatoms (Sig1). It was further discussed that the Pacific/Indonesian T. weissflogii species complex most likely evolved in the presence of gene flow and "local" environmental selection. Three hypotheses which potentially can explain the genetic diversification/similarity between the Atlantic T. weissflogii samples and the isolate obtained off the coast of California were proposed and discussed. PMID- 20018564 TI - Being reasonable: supporting disabled nursing students in practice. AB - AIM: To analyse recurring adjustments made in practice settings and the support strategies put in place to enable disabled students to achieve the levels of proficiency required on pre-registration nursing programmes. BACKGROUND: Legislative and regulatory changes in the UK require higher education institutions to make reasonable adjustments for disabled students whose needs must be considered and adjustments made before their programmes of study begin. The student practice learning advisor's (SPLA) primary role is to support disabled students and to operationalise recommended adjustments in practice. METHOD: An evaluative case study design was employed to analyse the work of the SPLA over 12 months using progression data, individual interviews and reflective accounts. FINDINGS: The evaluation illustrates the need for support throughout the student's programme which appears to reach a peak in the final year. Disabled students required 20% more contact time than their non-disabled peers. Operationalising adjustments requires attention to inter-disciplinary, practical and communication considerations. CONCLUSION: Implementing adjustments in practice requires a multi-disciplinary approach in order to support disabled students, and their mentors and to enable the development and application of coping strategies to overcome potential restrictions. The SPLA provides an essential role in ensuring opportunities for the disabled student to succeed are maximised. PMID- 20018565 TI - Common variable immunodeficiency and granulomatosis treated with infliximab. PMID- 20018566 TI - Pharmacological treatment of neuropathic facial pain in the dutch general population. AB - Few drugs are registered for treatment of neuropathic facial pain (NFP), and not much is known about treatment choices for NFP in daily practice. Patients with NFP were identified in the IPCI-database with longitudinal electronic general practitioner (GP) records. We described prescription patterns of pain medication following first symptoms. Off-label, off-guideline use, failure and reasons for failure were assessed. Failure was defined as treatment switch, exacerbation, adverse event, or invasive treatment for NFP. Of 203 NFP cases, 160 (79%) received pharmacological pain treatment. Most patients (90%) were initially treated by a GP with anti-epileptic drugs (55%) or NSAIDs (16%) as monotherapy. The median treatment delay was 0 days (range 0 to 2,478 days). Adverse events were experienced by 16 (10%) of patients. Sixty-two percent of first prescriptions were in adherence to guidelines and 59% were considered on-label while 34% of prescriptions were both off-label and off-guideline. Of the first therapy, 38% failed within 3 months. The median duration until failure was 251 days. General practitioners usually are the first to treat NFP. They usually prescribe drugs licensed for NFP and according to guidelines, but the extent of off-label use is substantial. Initial treatment often failed within a short period after starting therapy. PERSPECTIVE: This drug-utilization study describes the pharmacological treatment of different forms of neuropathic facial pain in daily practice. Although treatment is mostly initiated rapidly by general practitioners in a correct way, it often contains off-label or off-guideline medication. Failure of the initial treatment is common and occurs rapidly as well. PMID- 20018567 TI - Effects of anodal transcranial direct current stimulation on chronic neuropathic pain in patients with multiple sclerosis. AB - Neuropathic pain in patients with MS is frequent and is associated with a great interference with daily life activities. In the present study, we investigated whether anodal transcranial direct current stimulation (tDCS) may be effective in reducing central chronic pain in MS patients. Patients received sham tDCS or real tDCS in a 5-day period of treatment in a randomized, double blind, sham controlled study. Pain was measured using visual analog scale (VAS) for pain and the short form McGill questionnaire (SF-MPQ). Quality of life was measured using the Multiple Sclerosis Quality of Life-54 scale (MSQoL-54). Depressive symptoms and anxiety were also evaluated as confounding factors using the Beck Depression Inventory (BDI) and VAS for anxiety. Evaluations were performed at baseline, immediately after the end of treatment, and once a week during a 3-week follow-up period. Following anodal but not sham tDCS over the motor cortex, there was a significant pain improvement as assessed by VAS for pain and McGill questionnaire, and of overall quality of life. No depression or anxiety changes were observed. Our results show that anodal tDCS is able to reduce pain-scale scores in MS patients with central chronic pain and that this effect outlasts the period of stimulation, leading to long-lasting clinical effects. PERSPECTIVE: This article presents a new, noninvasive therapeutic approach to chronic, central neuropathic pain in multiple sclerosis, poorly responsive to current conventional medications. tDCS is known to cause long-lasting changes of neuronal excitability at the site of stimulation and in the connected areas in healthy subjects. This led us to hypothesize that pain decrease may be the result of functional plastic changes in brain structures involved in the pathogenesis of chronic neuropathic pain. PMID- 20018568 TI - Outcomes of lumbar discectomy. PMID- 20018569 TI - Atomoxetine changes rat's HR response to stress from tachycardia to bradycardia via alterations in autonomic function. AB - Atomoxetine is a central norepinephrine reuptake inhibitor used to treat attention deficit hyperactivity disorder. We tested the effects of atomoxetine upon the heart rate (HR) and mean arterial blood pressure (mBP) response to aversive conditioning. In Protocol 1 the mBP and HR responses to a stress (15s tone followed by shock) were tested in 8 Sprague-Dawley rats given saline pretreatment for 3 days; the rats' responses were then tested for 3 additional days following atomoxetine (1mg/kg, sc). Atomoxetine decreased (p<0.05) baseline mBP from 128+/-11 mm Hg (mean+/-SD) to 117+/-19 mm Hg; baseline HR slowed from 380+/-23 bpm to 351+/-21 bpm. The mBP increase to acute stress was similar after saline vs. after drug, but the peak was attained more slowly. After atomoxetine HR tended to slow during stress rather than accelerate. In Protocol 2 the cardiovascular responses were tested (n=6) for 3 days post-saline and for 3 days after a higher dose of atomoxetine (2mg/kg, sc). The average HR acceleration during the last 10s of the stress after saline (+7.5+/-14.7 bpm) was replaced by a HR slowing (-6.2+/-10.5 bpm). We conclude that drug treatment (a) decreases baseline sympathetic tone and/or elevates cardiac parasympathetic tone; (b) slows sympathetic arousal to acute stress without changing its magnitude; and, (c) enables the emergence of elevated parasympathetic tone during the stress. These autonomic consequences are consistent with atomoxetine's anxiolytic and transient sedative effects. PMID- 20018570 TI - History and perspectives of bioanalytical methods for chemical warfare agent detection. AB - This paper provides a short historical overview of the development of bioanalytical methods for chemical warfare (CW) agents and their biological markers of exposure, with a more detailed overview of methods for organophosphorus nerve agents. Bioanalytical methods for unchanged CW agents are used primarily for toxicokinetic/toxicodynamic studies. An important aspect of nerve agent toxicokinetics is the different biological activity and detoxification pathways for enantiomers. CW agents have a relatively short lifetime in the human body, and are hydrolysed, metabolised, or adducted to nucleophilic sites on macromolecules such as proteins and DNA. These provide biological markers of exposure. In the past two decades, metabolites, protein adducts of nerve agents, vesicants and phosgene, and DNA adducts of sulfur and nitrogen mustards, have been identified and characterized. Sensitive analytical methods have been developed for their detection, based mainly on mass spectrometry combined with gas or liquid chromatography. Biological markers for sarin, VX and sulfur mustard have been validated in cases of accidental and deliberate human exposures. The concern for terrorist use of CW agents has stimulated the development of higher throughput analytical methods in support of homeland security. PMID- 20018571 TI - HPLC method for determination of fluorescence derivatives of cortisol, cortisone and their tetrahydro- and allo-tetrahydro-metabolites in biological fluids. AB - 11Beta-hydroxysteroid dehydrogenase isoform 2 (11beta-HSD2) is responsible for conversion of cortisol (F) to inactive cortisone (E). Disturbance of its activity can cause hypertension. To estimate 11beta-HSD2 activity, besides F and E, their tetrahydro- (THF, THE) as well allo-tetrahydro- (allo-THF, allo-THE) metabolites should be determined. This study describes HPLC-FLD method for the quantitative determination of endogenous glucocorticoids (GCs) in plasma and urine (total and free) and their metabolites in urine. Following extraction at pH 7.4 using dichloromethane, GCs (F, E, THF, allo-THF, THE, allo-THE and internal standard- prednisolone) were derivatized with 9-anthroyl nitrile and purified by SPE using C(18) cartridges. The enzymatic hydrolysis of conjugated steroids was provided using beta-glucuronidase. The influence of organic bases on 9-AN derivatization of steroids was investigated. The best yield of the derivatization was obtained in presence of the mixture of 10.0% triethylamine (TEA) and 0.1% quinuclidine (Q). Chromatographic separation was accomplished in the Chromolith RP-18e monolithic column. The elaborated method was validated. Calibration curves were linear in the ranges: for F, E and THF 5.0-1000.0 ng mL(-1), for allo-THF and THE + allo-THE 10.0-1000.0 ng mL(-1). LOD (S/N=3:1) for all analytes amounted 3.0 ng mL(-1). Recoveries of GCs exceeded 90%. The method was precise and accurate, intra- and inter-day precision were 3.0-12.1% and 9.2-14.0%, respectively. Accuracy ranged from 0.2 to 15.1%. The method was applied for estimating endogenous GCs in plasma and urine. Plasma levels of F and E were in the ranges: 133.0-174.5 ng mL(-1) and 17.4-35.9 ng mL(-1), respectively. Free urinary steroids were in the ranges: 12.0-54.1 microg/24 h (UFF) and 37.8-76.2 microg/24 h (UFE). The ratio of (THF + allo-THF)/(THE + allo-THE) amounted from 1.01 to 1.23. The obtained results confirmed utility of the elaborated method in the assessment of 11beta-HSD2 activity in man. PMID- 20018572 TI - Enantiomeric determination of tramadol and O-desmethyltramadol in human plasma by fast liquid chromatographic technique coupled with mass spectrometric detection. AB - A rapid and sensitive method using liquid chromatography-tandem mass spectrometry (LC-MS/MS) for enantiomeric determination of tramadol and its primary phase metabolite O-desmethyltramadol in human plasma has been developed. Tramadol hydrochloride-(13)C, d(3), was used as an isotopic labeled internal standard for quantification. The method involves a simple solid phase extraction. The analytes and internal standard were separated on Lux Cellulose-2 packed with cellulose tris(3-chloro-4-methylphenylcarbamate) using isocratic elution with hexane/isopropanol/diethylamine (90:10:0.1, v/v/v) at a flow rate of 1.3 mL/min. The APCI positive ionization mass spectrometry was used with multiple reaction monitoring of the transitions at m/z 264.2-->58.2 for tramadol, m/z 250.1-->58.2 for O-desmethyltramadol and m/z 268.2-->58.2 for internal standard. Linearity was achieved between 1-800 ng/mL and 1-400 ng/mL (R(2) > or = 0.999) for each enantiomer of tramadol and O-desmethyltramadol, respectively. Intra-day accuracies ranged among 98.2-102.8%, 97.1-109.1% and 97.4-102.9% at the lower, intermediate, and high concentration for all analytes, respectively. Inter-day accuracies ranged among 95.5-104.1%, 99.2-104.7%, and 94.2-105.6% at the lower, intermediate, and high concentration for all analytes, respectively. This assay was successfully used to determine the concentration of enantiomers of tramadol and O-desmethyltramadol in a pharmacogenetic study. PMID- 20018573 TI - Toll-like receptor 4 participates in gastric mucosal protection through Cox-2 and PGE2. AB - AIM: To elucidate the role of Toll-like receptors (TLRs) in gastric cytoprotection after ethanol injury. METHODS: C57BL/6J, C3H/HeOuJ and C3H/HeJ mice were used. All mice were killed 4h after ethanol administration. TLR4, cyclooxygenase-2 (Cox-2) and prostaglandin E(2) (PGE(2)) expression were measured by immunohistochemistry, western blotting and enzyme-linked immunosorbent assay (ELISA) separately. RESULTS: The expression of TLR4 increased in C57BL/6J mice stomach 4h after ethanol injury. The cells expressing TLR4 included Cox-2 expressing cells and macrophages. The injury in C3H/HeJ mice was more severe than in C3H/HeOuJ mice 4h after ethanol injury. The expression of Cox-2 and PGE(2) only increased in C3H/HeOuJ mice. The number of macrophages and the expression of macrophage-inflammatory protein-2 (MIP-2) also increased only in C3H/HeOuJ mice. CONCLUSION: TLR4 signal is activated in mice stomach 4h after ethanol injury. The protective effects of TLR4 signalling are mediated through the induction of Cox-2 expression and the production of PGE(2). PMID- 20018574 TI - ERCP-induced and non-ERCP-induced acute pancreatitis: Two distinct clinical entities with different outcomes in mild and severe form? AB - BACKGROUND: Acute pancreatitis is a complication of endoscopic retrograde cholangio-pancreatography. Aim of the study was to compare endoscopic retrograde cholangio-pancreatography-related acute pancreatitis with attacks caused by other factors. METHODS: A series of consecutive patients with non-endoscopic retrograde cholangio-pancreatography-related acute pancreatitis referred to our hospital in 2007-2008 were examined retrospectively, and compared with the same number of patients with post-endoscopic retrograde cholangio-pancreatography acute pancreatitis done in the same institution. Both groups comprised 116 patients and were comparable for mean age, sex, and body mass index. Duration of abdominal pain, pancreatic enzyme elevation, hospital stay, and type of analgesia administered were retrieved. RESULTS: There were no differences between the groups as regards the severity of pancreatitis, mortality rate and hospitalisation, although mortality was double in severe post-endoscopic retrograde cholangio-pancreatography acute pancreatitis. In the mild acute pancreatitis cases, serum amylase fell 50% from the peak in a mean of 46.4h (range 24-72) in group 1 and 38.9h (range 24-72) in group 2 (p<0.001). The peak amylase serum level halved within 48h in 73.6% of cases with non-endoscopic retrograde cholangio-pancreatography-related acute pancreatitis, and in 92% of patients with endoscopic retrograde cholangio-pancreatography-related acute pancreatitis (p<0.001). CONCLUSIONS: Non-endoscopic retrograde cholangio pancreatography- and endoscopic retrograde cholangio-pancreatography-induced pancreatitis did not differ as regards severity, hospital stay or mortality; in mild pancreatitis, serum amylase halved significantly sooner in post-endoscopic retrograde cholangio-pancreatography cases. PMID- 20018575 TI - No beneficial effects of amantadine in treatment of chronic hepatitis C patients. AB - BACKGROUND: Benefit of adding amantadine to antiviral therapy for hepatitis C is controversial. AIMS: We aimed to examine whether such policy enhances sustained viral response in treatment-naive patients. METHODS: 297 naive hepatitis C patients were randomized for treatment with amantadine 200mg or placebo, combined with weight-based ribavirin and 12-day high-dose interferon alpha-2b induction therapy, followed by PEG-interferon alpha-2b (1.5 microg/kg/week up to 26 weeks and thereafter, 1.0 microg/kg/week until week 52). Treatment was discontinued if hepatitis C virus (HCV) RNA was positive at week 24. RESULTS: 49% of patients were (former) drug users. Genotype 1 occurred in 45%, high viral load in 70% and severe fibrosis/cirrhosis in 32%, without differences between amantadine or placebo groups. 90 patients prematurely discontinued treatment, mainly because of grade 3 or 4 toxicity. Intention-to-treat analysis revealed sustained viral response in 47% and 51% of amantadine and placebo groups (p=0.49). Amantadine did not enhance sustained viral response in patients with genotype 1 or high viral load nor did it improve primary non-response, breakthrough or relapse rates. Genotype non-1 and lower pre-treatment gamma GT levels were independent predictors for sustained viral response. CONCLUSION: Adding amantadine to antiviral therapy of previously untreated chronic hepatitis C patients has no beneficial effects. PMID- 20018576 TI - [Succinic semialdehyde dehydrogenase deficiency: decrease in 4-OH-butyric acid levels with low doses of vigabatrin]. AB - Succinic semialdehyde dehydrogenase deficiency (gamma-hydroxybutyric aciduria) is a rare neurometabolic disease caused by a deficiency in gamma-aminobutyric degradation, resulting in an increase in gamma-hydroxybutyric acid in biological fluids. The clinical spectrum is heterogeneous, including a variety of neurological manifestations and psychiatric symptoms. The treatment usually used is vigabatrin, but its clinical efficacy is under discussion. We present two affected siblings. The older brother was examined when he was 2.5 years old due to psychomotor and developmental delay, disturbances in motor coordination, axial hypotonia and language disability. His younger brother had mild axial hypotonia when 5 months old. Metabolic studies demonstrated a high plasma and urine concentration of gamma-hydroxybutyric acid. Mutation analysis of the gene ALDH5A1 confirmed the disease. After 1 year of treatment with low-doses of vigabatrin of the older patient, a decrease in gamma-hydroxybutyric acid plasma levels and a slow clinical improvement were observed. PMID- 20018577 TI - [Is it worth knowing the characteristics of inpatient paediatric parenteral nutrition?]. AB - OBJECTIVE: To describe the characteristics of inpatient parenteral nutrition during 2008 in a tertiary level paediatric hospital. To compare this practice with recent standards. MATERIAL AND METHODS: Longitudinal, descriptive, and retrospective study of patients under 16 years receiving parenteral nutrition in 2008. Demographic data as well as data of the nutritional support were recorded. RESULTS: A total of 120 children received parenteral nutrition, of whom 63 were under 2 years, 30 between 3 and 7 years, and 27 older than 7 years. They accounted for 1.6% of all the inpatients during that period. Most frequent indication was gastrointestinal surgery (35.5%), followed by non-surgical gastrointestinal diseases (19.8%). The average composition in the first day of parenteral nutrition was: 63.1kcal/kg (SD: 21.3); volume, 76.6ml/kg (SD: 28.4); carbohydrates 9.6g/kg/d (SD: 3.2); amino acids 2.1g/kg/d (SD: 0.6), and lipids 1.9g/kg/d (SD: 1). The mean duration of parenteral support was 10.5 days (SD: 9.8). Internal jugular vein (34.7%) and femoral vein were the most common intravenous access. The main reason to discontinue parenteral nutrition was transition to oral/enteral feedings. The complication rate was 27.3%. CONCLUSIONS: There is an interest in evaluating standard practice in parenteral nutrition in order to develop improvement strategies. Infants and toddlers are the principal candidates for parenteral nutrition. Standardisation is one way to decrease errors in its preparation and prescription. PMID- 20018578 TI - [Quality of life of preterm infants and admissions due to respiratory infections]. AB - INTRODUCTION: Study of the association between lower respiratory tract infection hospitalisations and health related quality of life (HRQoL) in preterm infants of 32-35 weeks of gestational age. METHODS: Survey study nested into a prospective follow-up cohort study of preterm infants (FLIP-2). During the last FLIP-2 visit, 216 preterm-parent pairs were interviewed. The structured questionnaire included measures of HRQoL (QUALIN modified scale for the infant, and SF-12 for the parent, and Visual scales for both), caregiver overload (Zarit modified scale and indirect measurements). RESULTS: From October 2006 to March 2007 (RSV season), there were 71 respiratory hospitalisations (33%). Triplets and infants living in homes with >5 inhabitants were most likely to be hospitalised. Parents of hospitalised children were most likely, to have more and longer times off work for child care (47% vs. 18%), to have higher overload, and to obtain lower values in the physical dimension of SF-12. Multiple regression model associated infant HRQoL with higher gestational age, having 0-3 year-old siblings, being recommended palivizumab and had received it, lower caregiver overload, higher caregiver mental HRQoL and no absence from work for child care. CONCLUSIONS: Although respiratory hospitalisations were not associated with infant HRQoL, caregivers' HRQoL and overload were. Preterm infant HRQoL is associated with their caregivers' HRQoL and overload, and with receiving RSV prophylaxis when their risk profile recommends it. PMID- 20018579 TI - Acute upper limb ischaemia following chest wall reconstruction--a word of caution. AB - Irradiation arteriopathy, although rare, is a recognised entity following radiotherapy for the management of advanced cancers. It is known to manifest in varied forms and can be chronic and progressive. We present a case of an acute upper limb ischaemia following deep inferior epigastric artery perforator (DIEP) free flap reconstruction for a chest wall defect in a patient who had previously undergone radiotherapy 25 years back for carcinoma of the breast. It is envisaged that the axillary artery was thrombosed secondary to radiation arteriopathy and the collaterals that had developed were probably damaged during debridement of the irradiated unhealthy tissue prior to free flap reconstruction. A cross-limb vascular graft tunnelled under the flap restored the perfusion to the limb. We propose that the possibility of radiation arteriopathy should be considered and a preoperative arteriogram should be performed prior to planning any major surgical reconstruction in patients having undergone irradiation to major vascular beds. PMID- 20018580 TI - Secondary surgery in paediatric facial paralysis reanimation. AB - Ninety-two children, the entire series of paediatric facial reanimation by a single surgeon over thirty years, are presented. The objective is to analyse the incidence and value of secondary revisions for functional and aesthetic refinements following the two main stages of reanimation. The reconstructive strategy varied according to the denervation time, the aetiology, and whether the paralysis was uni- or bilateral, complete or partial. Irrespective of these variables, 89% of the patients required secondary surgery. Post-operative videos were available in seventy-two cases. Four independent observers graded patients' videos using a scale from poor to excellent. The effect of diverse secondary procedures was measured computing a mean-percent-gain score. Statistical differences between treatment groups means were tested by the t-test and one-way ANOVA. Two-thirds of the corrective and ancillary techniques utilized granted significantly higher mean-scores post-secondary surgery. A comparison of pre- and post-operative data found valuable improvements in all three facial zones after secondary surgery. In conclusion, inherent to dynamic procedures is the need for secondary revisions. Secondary surgery builds in the potential of reanimation surgery, effectively augmenting functional faculties and aesthesis. PMID- 20018581 TI - Intraoperative diagnostic cytology of intra-parotid lymph nodes. PMID- 20018582 TI - Advanced midwifery practice or advancing midwifery practice? AB - Advanced midwifery practice is a controversial notion in midwifery, particularly at present in Australia. The proposed changes in legislation around access to the publicly funded Medical Benefits Scheme (MBS) and the Pharmaceutical Benefits Scheme (PBS) in 2009-2010 have meant that the issue of advanced midwifery practice has again taken prominence. Linking midwifery access to MBS and PBS to a safety and quality framework that includes an 'advanced midwifery credentialling framework' is particularly challenging. The Haxton and Fahy paper in the December 2009 edition of Women and Birth is timely as it enables a reflection upon these issues and encourages debate and discussion about exactly what is midwifery, what are we educating our students for and is working to the full scope of practice practising at advanced level? This paper seeks to address some of these questions and open up the topic for further debate. PMID- 20018583 TI - The use of dry powder inhaler devices by elderly patients suffering from chronic obstructive pulmonary disease. AB - Twenty-five COPD patients, aged 65years or above, were recruited to test their ability to use dry powder inhaler Handihaler (Boeringher-Ingelheim) and Aerolizer (Novartis). The results of a score created to evaluate the inhalation technique were compared with age, MMSE, Barthel Index, FEV(1), maximum inspiratory and expiratory pressures, and peak inspiratory flow (PIF). RESULTS: Dry powder inhalers were correctly used by 60% of the patients (15 out of 25). Among the capable ones, 13 out of 15 were aged less than 80 years (p< or =0.02), 13 out of 15 had a maximum inspiratory pressure greater or equal to 53cm H(2)O (p< or =0.001) and a PIF greater or equal to 120l/min (p< or =0.05). All skilled patients had a minimum MMSE of 25 (p< or =0.001). CONCLUSION: In a geriatric population, age, the decrease of maximum inspiratory pressure and PIF as well as cognitive functions, limit the use of dry powder inhalers. PMID- 20018585 TI - The rough-terrain problem: accurate foot targeting as a function of visual information regarding target location. AB - The authors examined step-cycle regulation to accurately land on a single target. They also examined the effect of decreasing and increasing visual information regarding target location. Visual information was decreased with goggles that obstructed the lower visual field, removing information of the target and foot relative to target in the two steps before the target. Visual information was increased by adding 4 vertical poles (2.1 m tall) around the target location. A total of 14 participants landed with 1 foot on a flat target placed halfway down a walkway and continued walking. During target approach, step length variability increased and foot placement variability decreased. The final stride onto the target was longer and smoother than the penultimate stride, which may reflect that modifications were made earlier to reduce modifications needed in the final stride. Foot-target accuracy was reduced by lower visual-field obstruction. In the steps preceding the target, the presence of poles describing target location modified foot-placement variability, stride length, swing-trajectory smoothness, and head angle. However, foot-target accuracy was not modified by increased visual information regarding target location. That is, the presence of poles modified how the task was performed but did not alter the outcome. Therefore, view of the foot relative to the target is more important than view of target location in the control of a foot-targeting task. PMID- 20018584 TI - [Allogeneic cord blood transplantation for adults: current results and future development]. AB - Hematopoietic stem cell transplantation is the choice treatment of many hematopoietic disorders. However, there is still no related or HLA-matched unrelated donor for one-third of the patients. Cord blood, which contains a lot of hematopoietic progenitors immunologically naive, represents not only an interesting alternative as hematopoietic stem cell source but also allows more HLA incompatibilities than the other sources. Promising results in children lead to develop cord blood transplantation in adults first of all in hematopoietic malignancies (acute leukemias and lymphoid diseases) and in a second time, in non malignant diseases such as aplastic anaemia. The main problems for the development of this new strategy in adults are the poor number of cells per unit, the delay for hematopoietic recovery in comparison with other hematopoietic stem cell sources and, consequently higher transplant related mortality. In order to improve the results in adults, new strategies emerged. Double cord blood transplantation, expansion methods and intra-bone injection of the graft will be reviewed here, as well as alternative transplantation strategies such as non myeloablative conditioning. PMID- 20018586 TI - Identifying leading joint strategies in a bimanual coordination task: does coordination stability depend on leading joint strategy? AB - The authors sought to determine if principles of the Leading Joint Hypothesis, when applied to a multijoint, bimanual coordination task, could provide insight into the contribution of intralimb dynamics to interlimb coordination. Participants repetitively traced ellipse templates with an isodirectional motion of the end effectors (both hands moving counterclockwise [CCW]) at two cycling frequencies. Ellipse templates were oriented either tilted right or tilted left, yielding a total of 4 left arm-right arm leading-joint combinations. Analysis of torque sign and impulse data indicated that the 4 ellipse-tracing conditions resulted in 4 distinct left arm-right arm leading-joint combinations: 2 conditions with similar leading joints and 2 conditions with different leading joints. Isodirectional CCW ellipse tracing was more stable when produced with similar leading joints compared with when produced with different leading joints. The authors discuss results within the context of intralimb control contributions to the stability of interlimb coordination patterns. PMID- 20018587 TI - Variations in linear and nonlinear postural measurements under achilles tendon vibration and unstable support-surface conditions. AB - Reduced support-surface stability has been shown to attenuate the effect of Achilles tendon vibration on backward body displacement. In the present study, 20 participants performed a quiet, upright standing task on a stable and sway referenced support, with and without vibration. The authors calculated equilibrium scores (ES), approximate entropy (ApEn), and mean and peak power spectral density frequencies of center-of-pressure variations. It was found that ES values decreased with the addition of vibration and in the sway-referenced support condition. ApEn values decreased with the addition of vibration but only with a stable support. Conversely, mean and peak frequencies increased with the addition of vibration, independent of support stability. These results suggest that the role of ankle proprioceptive input changes depending on support-surface characteristics and demonstrate the value of using both linear and nonlinear measures of postural sway. PMID- 20018588 TI - An electronic clinical decision support tool to assist primary care providers in cardiovascular disease risk management: development and mixed methods evaluation. AB - BACKGROUND: Challenges remain in translating the well-established evidence for management of cardiovascular disease (CVD) risk into clinical practice. Although electronic clinical decision support (CDS) systems are known to improve practitioner performance, their development in Australian primary health care settings is limited. OBJECTIVES: Study aims were to (1) develop a valid CDS tool that assists Australian general practitioners (GPs) in global CVD risk management, and (2) preliminarily evaluate its acceptability to GPs as a point-of care resource for both general and underserved populations. METHODS: CVD risk estimation (based on Framingham algorithms) and risk-based management advice (using recommendations from six Australian guidelines) were programmed into a software package. Tool validation: Data from 137 patients attending a physician's clinic were analyzed to compare the tool's risk scores with those obtained from an independently programmed algorithm in a separate statistics package. The tool's management advice was compared with a physician's recommendations based on a manual review of the guidelines. Field test: The tool was then tested with 21 GPs from eight general practices and three Aboriginal Medical Services. Customized CDS-based recommendations were generated for 200 routinely attending patients (33% Aboriginal) using information extracted from the health record by a research assistant. GPs reviewed these recommendations during each consultation. Changes in CVD risk factor measurement and management were recorded. In-depth interviews with GPs were conducted. RESULTS: Validation testing: the tool's risk assessment algorithm correlated very highly with the independently programmed version in the separate statistics package (intraclass correlation coefficient 0.999). For management advice, there were only two cases of disagreement between the tool and the physician. Field test: GPs found 77% (153/200) of patient outputs easy to understand and agreed with screening and prescribing recommendations in 72% and 64% of outputs, respectively; 26% of patients had their CVD risk factor history updated; 73% had at least one CVD risk factor measured or tests ordered. For people assessed at high CVD risk (n = 82), 10% and 9%, respectively, had lipid-lowering and BP-lowering medications commenced or dose adjustments made, while 7% newly commenced anti-platelet medications. Three key qualitative findings emerged: (1) GPs found the tool enabled a systematic approach to care; (2) the tool greatly influenced CVD risk communication; (3) successful implementation into routine care would require integration with practice software, minimal data entry, regular revision with updated guidelines, and a self-auditing feature. There were no substantive differences in study findings for Aboriginal Medical Services GPs, and the tool was generally considered appropriate for use with Aboriginal patients. CONCLUSION: A fully integrated, self-populating, and potentially Internet-based CDS tool could contribute to improved global CVD risk management in Australian primary health care. The findings from this study will inform a large-scale trial intervention. PMID- 20018590 TI - Increasing sucrose uptake capacity of wheat grains stimulates storage protein synthesis. AB - Increasing grain sink strength by improving assimilate uptake capacity could be a promising approach toward getting higher yield. The barley (Hordeum vulgare) sucrose transporter HvSUT1 (SUT) was expressed under control of the endosperm specific Hordein B1 promoter (HO). Compared with the wild type, transgenic HOSUT grains take up more sucrose (Suc) in vitro, showing that the transgene is functional. Grain Suc levels are not altered, indicating that Suc fluxes are influenced rather than steady-state levels. HOSUT grains have increased percentages of total nitrogen and prolamins, which is reflected in increased levels of phenylalanine, tyrosine, tryptophan, isoleucine, and leucine at late grain development. Transcript profiling indicates specific stimulation of prolamin gene expression at the onset of storage phase. Changes in gene expression and metabolite levels related to carbon metabolism and amino acid biosynthesis suggest deregulated carbon-nitrogen balance, which together indicate carbon sufficiency and relative depletion of nitrogen. Genes, deregulated together with prolamin genes, might represent candidates, which respond positively to assimilate supply and are related to sugar-starch metabolism, cytokinin and brassinosteroid functions, cell proliferation, and sugar/abscisic acid signaling. Genes showing inverse expression patterns represent potential negative regulators. It is concluded that HvSUT1 overexpression increases grain protein content but also deregulates the metabolic status of wheat (Triticum aestivum) grains, accompanied by up-regulated gene expression of positive and negative regulators related to sugar signaling and assimilate supply. In HOSUT grains, alternating stimulation of positive and negative regulators causes oscillatory patterns of gene expression and highlights the capacity and great flexibility to adjust wheat grain storage metabolism in response to metabolic alterations. PMID- 20018592 TI - Gravity-induced modifications to development in hypocotyls of Arabidopsis tubulin mutants. AB - We investigated the roles of cortical microtubules in gravity-induced modifications to the development of stem organs by analyzing morphology and orientation of cortical microtubule arrays in hypocotyls of Arabidopsis (Arabidopsis thaliana) tubulin mutants, tua3(D205N), tua4(S178Delta), and tua6(A281T), cultivated under 1g and hypergravity (300g) conditions. Hypocotyls of tubulin mutants were shorter and thicker than the wild type even at 1g, and hypergravity further suppressed elongation and stimulated expansion. The degree of such changes was clearly smaller in tubulin mutants, in particular in tua6. Hypocotyls of tubulin mutants also showed either left-handed or right-handed helical growth at 1g, and the degree of twisting phenotype was intensified under hypergravity conditions, especially in tua6. Hypergravity induced reorientation of cortical microtubules from transverse to longitudinal directions in epidermal cells of wild-type hypocotyls. In tubulin mutants, especially in tua6, the percentage of cells with longitudinal microtubules was high even at 1g, and it was further increased by hypergravity. The twisting phenotype was most obvious at cells 10 to 12 from the top, where reorientation of cortical microtubules from transverse to longitudinal directions occurred. Moreover, the left-handed helical growth mutants (tua3 and tua4) had right-handed microtubule arrays, whereas the right-handed mutant (tua6) had left-handed arrays. There was a close correlation between the alignment angle of epidermal cell files and the alignment of cortical microtubules. Gadolinium ions, blockers of mechanosensitive ion channels (mechanoreceptors), suppressed the twisting phenotype in tubulin mutants under both 1g and 300 g conditions. Microtubule arrays in tubulin mutants were oriented more transversely by gadolinium treatment, irrespective of gravity conditions. These results support the hypothesis that cortical microtubules play an essential role in maintenance of normal growth phenotype against the gravitational force, and suggest that mechanoreceptors are involved in modifications to morphology and orientation of microtubule arrays by 1g gravity and hypergravity in tubulin mutants. PMID- 20018591 TI - Divalent metal ions in plant mitochondria and their role in interactions with proteins and oxidative stress-induced damage to respiratory function. AB - Understanding the metal ion content of plant mitochondria and metal ion interactions with the proteome are vital for insights into both normal respiratory function and the process of protein damage during oxidative stress. We have analyzed the metal content of isolated Arabidopsis (Arabidopsis thaliana) mitochondria, revealing a 26:8:6:1 molar ratio for iron:zinc:copper:manganese and trace amounts of cobalt and molybdenum. We show that selective changes occur in mitochondrial copper and iron content following in vivo and in vitro oxidative stresses. Immobilized metal affinity chromatography charged with Cu(2+), Zn(2+), and Co(2+) was used to identify over 100 mitochondrial proteins with metal binding properties. There were strong correlations between the sets of immobilized metal affinity chromatography-interacting proteins, proteins predicted to contain metal-binding motifs, and protein sets known to be oxidized or degraded during abiotic stress. Mitochondrial respiratory chain pathways and matrix enzymes varied widely in their susceptibility to metal-induced loss of function, showing the selectivity of the process. A detailed study of oxidized residues and predicted metal interaction sites in the tricarboxylic acid cycle enzyme aconitase identified selective oxidation of residues in the active site and showed an approach for broader screening of functionally significant oxidation events in the mitochondrial proteome. PMID- 20018594 TI - Aneuploidy causes tissue-specific qualitative changes in global gene expression patterns in maize. AB - Segmental aneuploidy refers to the relative excess or deficiency of specific chromosome regions. This condition results in gene dosage imbalance and often causes severe phenotypic alterations in plants and animals. The mechanisms by which gene dosage imbalance affects gene expression and phenotype are not completely clear. The effects of aneuploidy on the transcriptome may depend on the types of cells analyzed and on the developmental stage. We performed global gene expression profiling to determine the effects of segmental aneuploidy on gene expression levels in two different maize (Zea mays) tissues and a detailed analysis of expression of 30 genes affected by aneuploidy in multiple maize tissues. Different maize tissues varied in the frequency at which genes located outside of the aneuploid regions are positively or negatively regulated as well as in the degree of gene dosage compensation. Multiple genes demonstrated qualitative changes in gene expression due to aneuploidy, when the gene became ectopically expressed or completely silenced in aneuploids relative to wild-type plants. Our data strongly suggested that quantitative changes in gene expression at developmental transition points caused by variation in gene copy number progressed through tissue development and resulted in stable qualitative changes in gene expression patterns. Thus, aneuploidy in maize results in alterations of gene expression patterns that differ between tissues and developmental stages of maize seedlings. PMID- 20018593 TI - Metabolic networks: how to identify key components in the regulation of metabolism and growth. PMID- 20018595 TI - Gene expression analysis, proteomics, and network discovery. PMID- 20018596 TI - Hydrophobin fusions for high-level transient protein expression and purification in Nicotiana benthamiana. AB - Insufficient accumulation levels of recombinant proteins in plants and the lack of efficient purification methods for recovering these valuable proteins have hindered the development of plant biotechnology applications. Hydrophobins are small and surface-active proteins derived from filamentous fungi that can be easily purified by a surfactant-based aqueous two-phase system. In this study, the hydrophobin HFBI sequence from Trichoderma reesei was fused to green fluorescent protein (GFP) and transiently expressed in Nicotiana benthamiana plants by Agrobacterium tumefaciens infiltration. The HFBI fusion significantly enhanced the accumulation of GFP, with the concentration of the fusion protein reaching 51% of total soluble protein, while also delaying necrosis of the infiltrated leaves. Furthermore, the endoplasmic reticulum-targeted GFP-HFBI fusion induced the formation of large novel protein bodies. A simple and scalable surfactant-based aqueous two-phase system was optimized to recover the HFBI fusion proteins from leaf extracts. The single-step phase separation was able to selectively recover up to 91% of the GFP-HFBI up to concentrations of 10 mg mL( 1). HFBI fusions increased the expression levels of plant-made recombinant proteins while also providing a simple means for their subsequent purification. This hydrophobin fusion technology, when combined with the speed and posttranslational modification capabilities of plants, enhances the value of transient plant-based expression systems. PMID- 20018597 TI - Stimulation of nonselective amino acid export by glutamine dumper proteins. AB - Phloem and xylem transport of amino acids involves two steps: export from one cell type to the apoplasm, and subsequent import into adjacent cells. High affinity import is mediated by proton/amino acid cotransporters, while the mechanism of export remains unclear. Enhanced expression of the plant-specific type I membrane protein Glutamine Dumper1 (GDU1) has previously been shown to induce the secretion of glutamine from hydathodes and increased amino acid content in leaf apoplasm and xylem sap. In this work, tolerance to low concentrations of amino acids and transport analyses using radiolabeled amino acids demonstrate that net amino acid uptake is reduced in the glutamine secreting GDU1 overexpressor gdu1-1D. The net uptake rate of phenylalanine decreased over time, and amino acid net efflux was increased in gdu1-1D compared with the wild type, indicating increased amino acid export from cells. Independence of the export from proton gradients and ATP suggests that overexpression of GDU1 affects a passive export system. Each of the seven Arabidopsis (Arabidopsis thaliana) GDU genes led to similar phenotypes, including increased efflux of a wide spectrum of amino acids. Differences in expression profiles and functional properties suggested that the GDU genes fulfill different roles in roots, vasculature, and reproductive organs. Taken together, the GDUs appear to stimulate amino acid export by activating nonselective amino acid facilitators. PMID- 20018598 TI - MEF9, an E-subclass pentatricopeptide repeat protein, is required for an RNA editing event in the nad7 transcript in mitochondria of Arabidopsis. AB - RNA editing in plants alters specific nucleotides from C to U in mRNAs in plastids and in mitochondria. I here characterize the nuclear gene MITOCHONDRIAL EDITING FACTOR9 (MEF9) that is required for RNA editing of the site nad7-200 in the nad7 mitochondrial mRNA in Arabidopsis (Arabidopsis thaliana). The MEF9 protein belongs to the E subfamily of pentatricopeptide repeat proteins and unlike the three previously identified mitochondrial editing factors MEF1 and MEF11 in Arabidopsis and OGR1 in rice (Oryza sativa) does not contain a DYW C terminal domain. In addition, the E domain is incomplete, but seems to be functionally required, since one of the two independent EMS mutants encodes a MEF9 protein truncated by a stop codon at the beginning of the E domain. In both mutant plants premature stop codons in MEF9 inactivate RNA editing at site nad7 200. The homozygous mutant plants are viable and develop rather normally. The lack of RNA editing at site nad7-200 thus seems to be tolerated although this editing event is conserved in most plant species or the genomic sequence already codes for a T at this position, resulting in a generally conserved amino acid codon. PMID- 20018599 TI - The Laforin-like dual-specificity phosphatase SEX4 from Arabidopsis hydrolyzes both C6- and C3-phosphate esters introduced by starch-related dikinases and thereby affects phase transition of alpha-glucans. AB - The biochemical function of the Laforin-like dual-specific phosphatase AtSEX4 (EC 3.1.3.48) has been studied. Crystalline maltodextrins representing the A- or the B-type allomorph were prephosphorylated using recombinant glucan, water dikinase (StGWD) or the successive action of both plastidial dikinases (StGWD and AtPWD). AtSEX4 hydrolyzed carbon 6-phosphate esters from both the prephosphorylated A- and B-type allomorphs and the kinetic constants are similar. The phosphatase also acted on prelabeled carbon-3 esters from both crystalline maltodextrins. Similarly, native starch granules prelabeled in either the carbon-6 or carbon-3 position were also dephosphorylated by AtSEX4. The phosphatase did also hydrolyze phosphate esters of both prephosphorylated maltodextrins when the (phospho)glucans had been solubilized by heat treatment. Submillimolar concentrations of nonphosphorylated maltodextrins inhibited AtSEX4 provided they possessed a minimum of length and had been solubilized. As opposed to the soluble phosphomaltodextrins, the AtSEX4-mediated dephosphorylation of the insoluble substrates was incomplete and at least 50% of the phosphate esters were retained in the pelletable (phospho)glucans. The partial dephosphorylation of the insoluble glucans also strongly reduced the release of nonphosphorylated chains into solution. Presumably, this effect reflects fast structural changes that following dephosphorylation occur near the surface of the maltodextrin particles. A model is proposed defining distinct stages within the phosphorylation/dephosphorylation-dependent transition of alpha-glucans from the insoluble to the soluble state. PMID- 20018600 TI - Studies of the kinetic mechanism of maize endosperm ADP-glucose pyrophosphorylase uncovered complex regulatory properties. AB - ADP-glucose pyrophosphorylase catalyzes the synthesis of ADP-glucose (ADP-Glc) from Glc-1-phosphate (G-1-P) and ATP. Kinetic studies were performed to define the nature of the reaction, both in the presence and absence of allosteric effector molecules. When 3-phosphoglycerate (3-PGA), the putative physiological activator, was present at a saturating level, initial velocity studies were consistent with a Theorell-Chance BiBi mechanism and product inhibition data supported sequential binding of ATP and G-1-P, followed by ordered release of pyrophosphate and ADP-Glc. A sequential mechanism was also followed when 3-PGA was absent, but product inhibition patterns changed dramatically. In the presence of 3-PGA, ADP-Glc is a competitive inhibitor with respect to ATP. In the absence of 3-PGA--with or without 5.0 mm inorganic phosphate--ADP-Glc actually stimulated catalytic activity, acting as a feedback product activator. By contrast, the other product, pyrophosphate, is a potent inhibitor in the absence of 3-PGA. In the presence of subsaturating levels of allosteric effectors, G-1-P serves not only as a substrate but also as an activator. Finally, in the absence of 3-PGA, inorganic phosphate, a classic inhibitor or antiactivator of the enzyme, stimulates enzyme activity at low substrate by lowering the K(M) values for both substrates. PMID- 20018601 TI - A putative phosphatase, LSF1, is required for normal starch turnover in Arabidopsis leaves. AB - A putative phosphatase, LSF1 (for LIKE SEX4; previously PTPKIS2), is closely related in sequence and structure to STARCH-EXCESS4 (SEX4), an enzyme necessary for the removal of phosphate groups from starch polymers during starch degradation in Arabidopsis (Arabidopsis thaliana) leaves at night. We show that LSF1 is also required for starch degradation: lsf1 mutants, like sex4 mutants, have substantially more starch in their leaves than wild-type plants throughout the diurnal cycle. LSF1 is chloroplastic and is located on the surface of starch granules. lsf1 and sex4 mutants show similar, extensive changes relative to wild type plants in the expression of sugar-sensitive genes. However, although LSF1 and SEX4 are probably both involved in the early stages of starch degradation, we show that LSF1 neither catalyzes the same reaction as SEX4 nor mediates a sequential step in the pathway. Evidence includes the contents and metabolism of phosphorylated glucans in the single mutants. The sex4 mutant accumulates soluble phospho-oligosaccharides undetectable in wild-type plants and is deficient in a starch granule-dephosphorylating activity present in wild-type plants. The lsf1 mutant displays neither of these phenotypes. The phenotype of the lsf1/sex4 double mutant also differs from that of both single mutants in several respects. We discuss the possible role of the LSF1 protein in starch degradation. PMID- 20018602 TI - Systematic localization of the Arabidopsis core cell cycle proteins reveals novel cell division complexes. AB - Cell division depends on the correct localization of the cyclin-dependent kinases that are regulated by phosphorylation, cyclin proteolysis, and protein-protein interactions. Although immunological assays can define cell cycle protein abundance and localization, they are not suitable for detecting the dynamic rearrangements of molecular components during cell division. Here, we applied an in vivo approach to trace the subcellular localization of 60 Arabidopsis (Arabidopsis thaliana) core cell cycle proteins fused to green fluorescent proteins during cell division in tobacco (Nicotiana tabacum) and Arabidopsis. Several cell cycle proteins showed a dynamic association with mitotic structures, such as condensed chromosomes and the preprophase band in both species, suggesting a strong conservation of targeting mechanisms. Furthermore, colocalized proteins were shown to bind in vivo, strengthening their localization function connection. Thus, we identified unknown spatiotemporal territories where functional cell cycle protein interactions are most likely to occur. PMID- 20018604 TI - Apyrase (nucleoside triphosphate-diphosphohydrolase) and extracellular nucleotides regulate cotton fiber elongation in cultured ovules. AB - Ectoapyrase enzymes remove the terminal phosphate from extracellular nucleoside tri- and diphosphates. In Arabidopsis (Arabidopsis thaliana), two ectoapyrases, AtAPY1 and AtAPY2, have been implicated as key modulators of growth. In fibers of cotton (Gossypium hirsutum), transcript levels for GhAPY1 and GhAPY2, two closely related ectoapyrases that have high sequence similarity to AtAPY1 and AtAPY2, are up-regulated when fibers enter their rapid growth phase. In an ovule culture system, fibers release ATP as they grow, and when their ectoapyrase activity is blocked by the addition of polyclonal anti-apyrase antibodies or by two different small molecule inhibitors, the medium ATP level rises and fiber growth is suppressed. High concentrations of the poorly hydrolyzable nucleotides ATPgammaS and ADPbetaS applied to the medium inhibit fiber growth, and low concentrations of them stimulate growth, but treatment with adenosine 5'-O-thiomonophosphate causes no change in the growth rate. Both the inhibition and stimulation of growth by applied nucleotides can be blocked by an antagonist that blocks purinoceptors in animal cells, and by adenosine. Treatment of cotton ovule cultures with ATPgammaS induces increased levels of ethylene, and two ethylene antagonists, aminovinylglycine and silver nitrate, block both the growth stimulatory and growth inhibitory effects of applied nucleotides. In addition, the ethylene precursor, 1-aminocyclopropane-1-carboxylic acid, lowers the concentration of nucleotide needed to promote fiber growth. These data indicate that ectoapyrases and extracellular nucleotides play a significant role in regulating cotton fiber growth and that ethylene is a likely downstream component of the signaling pathway. PMID- 20018606 TI - Critical care aspects of malaria. AB - Malaria remains a major health problem in much of Asia and Africa. A steady number of cases of imported malaria are also seen in many countries of the developed world. Plasmodium falciparum malaria and to some extent malaria caused by other species of Plasmodium can lead to many complications such as acute respiratory distress syndrome (ARDS), cerebral malaria, acute renal failure, severe anemia, thrombocytopenia, and bleeding complications. About 10% of patients with severe malaria die, usually as a result of multiorgan dysfunction. Critical care physicians should be aware of the complications and management of severe malaria. There has been significant progress in the understanding of pathogenesis of severe malaria over the last decade. Effective management of severe malaria includes early suspicion, prompt diagnosis, early institution of appropriate antimalarial chemotherapy, and supportive care, preferably in an intensive care unit. In this article, we review the different manifestations of severe malaria as relevant to critical care physicians and discuss the principles of laboratory diagnosis and management. PMID- 20018605 TI - Characterization of the possible roles for B class MADS box genes in regulation of perianth formation in orchid. AB - To investigate sepal/petal/lip formation in Oncidium Gower Ramsey, three paleoAPETALA3 genes, O. Gower Ramsey MADS box gene5 (OMADS5; clade 1), OMADS3 (clade 2), and OMADS9 (clade 3), and one PISTILLATA gene, OMADS8, were characterized. The OMADS8 and OMADS3 mRNAs were expressed in all four floral organs as well as in vegetative leaves. The OMADS9 mRNA was only strongly detected in petals and lips. The mRNA for OMADS5 was only strongly detected in sepals and petals and was significantly down-regulated in lip-like petals and lip like sepals of peloric mutant flowers. This result revealed a possible negative role for OMADS5 in regulating lip formation. Yeast two-hybrid analysis indicated that OMADS5 formed homodimers and heterodimers with OMADS3 and OMADS9. OMADS8 only formed heterodimers with OMADS3, whereas OMADS3 and OMADS9 formed homodimers and heterodimers with each other. We proposed that sepal/petal/lip formation needs the presence of OMADS3/8 and/or OMADS9. The determination of the final organ identity for the sepal/petal/lip likely depended on the presence or absence of OMADS5. The presence of OMADS5 caused short sepal/petal formation. When OMADS5 was absent, cells could proliferate, resulting in the possible formation of large lips and the conversion of the sepal/petal into lips in peloric mutants. Further analysis indicated that only ectopic expression of OMADS8 but not OMADS5/9 caused the conversion of the sepal into an expanded petal-like structure in transgenic Arabidopsis (Arabidopsis thaliana) plants. PMID- 20018603 TI - Novel bifunctional nucleases, OmBBD and AtBBD1, are involved in abscisic acid mediated callose deposition in Arabidopsis. AB - Screening of the expressed sequence tag library of the wild rice species Oryza minuta revealed an unknown gene that was rapidly and strongly induced in response to attack by a rice fungal pathogen (Magnaporthe oryzae) and an insect (Nilaparvata lugens) and by wounding, abscisic acid (ABA), and methyl jasmonate treatments. Its recombinant protein was identified as a bifunctional nuclease with both RNase and DNase activities in vitro. This gene was designated OmBBD (for O. minuta bifunctional nuclease in basal defense response). Overexpression of OmBBD in an Arabidopsis (Arabidopsis thaliana) model system caused the constitutive expression of the PDF1.2, ABA1, and AtSAC1 genes, which are involved in priming ABA-mediated callose deposition. This activation of defense responses led to an increased resistance against Botrytis cinerea. atbbd1, the knockout mutant of the Arabidopsis ortholog AtBBD1, was susceptible to attack by B. cinerea and had deficient callose deposition. Overexpression of either OmBBD or AtBBD1 in atbbd1 plants complemented the susceptible phenotype of atbbd1 against B. cinerea as well as the deficiency of callose deposition. We suggest that OmBBD and AtBBD1 have a novel regulatory role in ABA-mediated callose deposition. PMID- 20018607 TI - The accuracy of the central venous blood gas for acid-base monitoring. AB - BACKGROUND: Routine use of central venous blood gases (VBGs) may reduce complications from prolonged arterial cannulation. We investigated the reliability of the VBG as a substitute for arterial blood gas (ABG) in multiple care settings. METHODS: We developed a VBG adjustment rule of ABG pH = VBG pH + 0.05, ABG CO(2) = VBG PCO(2) -5 mm Hg from prior studies and validated this relationship with simultaneous venous and arterial blood obtained from 187 medical/surgical intensive care, cardiac catheterization laboratory, and coronary care unit patients with central venous access. RESULTS: The overall accuracy of a normal adjusted VBG (aVBG) to predict a normal ABG was 90%. After adjustment, the mean systematic difference (bias) between ABG and VBG pH decreased from 0.035 +/- 0.02 to -0.015 +/- 0.02 and PCO(2) bias decreased from -4.5 +/- 3.5 to 0.5 +/- 3.5. Intraclass correlation coefficients for agreement improved after applying the adjustment rule to venous pH (from 0.84 to 0.93, P < .001) and PCO(2) (from 0.66 to 0.84, P < .001). Overall diagnostic accuracy of VBG improved from 45% to 74% after adjustment. Multiple logistic regression demonstrated that the factor independently associated with discrepancy between VBG and ABG diagnoses was an abnormal aVBG (OR 6.8, 95% CI 2.8-16.5). CONCLUSIONS: Because of the high agreement between a normal aVBG with a normal ABG and the small bias between these tests, we recommend use of the adjusted central VBG. PMID- 20018608 TI - A score to predict early risk of recurrence after ischemic stroke. AB - BACKGROUND: There is currently no instrument to stratify patients presenting with ischemic stroke according to early risk of recurrent stroke. We sought to develop a comprehensive prognostic score to predict 90-day risk of recurrent stroke. METHODS: We analyzed data on 1,458 consecutive ischemic stroke patients using a Cox regression model with time to recurrent stroke as the response and clinical and imaging features typically available to physician at admission as covariates. The 90-day risk of recurrent stroke was calculated by summing up the number of independent predictors weighted by their corresponding beta-coefficients. The resultant score was called recurrence risk estimator at 90 days or RRE-90 score (available at: http://www.nmr.mgh.harvard.edu/RRE-90/). RESULTS: Sixty recurrent strokes (54 had baseline imaging) occurred during the follow-up period. The risk adjusted for time to follow-up was 6.0%. Predictors of recurrence included admission etiologic stroke subtype, prior history of TIA/stroke, and topography, age, and distribution of brain infarcts. The RRE-90 score demonstrated adequate calibration and good discrimination (area under the ROC curve [AUC] = 0.70-0.80), which was maintained when applied to a separate cohort of 433 patients (AUC = 0.70-0.76). The model's performance was also maintained for predicting early (14 day) risk of recurrence (AUC = 0.80). CONCLUSIONS: The RRE-90 is a Web-based, easy-to-use prognostic score that integrates clinical and imaging information available in the acute setting to quantify early risk of recurrent stroke. The RRE-90 demonstrates good predictive performance, suggesting that, if validated externally, it has promise for use in creating individualized patient management algorithms and improving clinical practice in acute stroke care. PMID- 20018609 TI - Here comes the sun?: a new online weather forecast for early stroke recurrence. PMID- 20018610 TI - The strength of B cell interaction with antigen determines the degree of IgM polymerization. AB - The induction of variable disulfide polymerization of IgM in the trout (Oncorhynchus mykiss) and its effect on its half-life were examined. An association between greater Ab affinity and increased disulfide polymerization was first indicated by the observation of this increased IgM disulfide polymerization during the process of affinity maturation. A direct association between Ab affinity and disulfide polymerization was then established by the fractionation of individual sera into high- and low-affinity subpopulations, which also resulted in the partitioning of high and low degrees of disulfide polymerization. The ability of high-affinity B cells to produce more highly polymerized Abs upon Ag induction was demonstrated by in vitro Ag-driven selection. Low Ag concentrations, which elicited only high-affinity Abs, also possessed the highest degree of polymerization, whereas higher concentrations of Ag elicited a broader array of Ab affinities, yielding a lower average affinity and degree of polymerization. Half-life studies revealed that the high-affinity, highly polymerized Abs possessed longer half-lives than the lower-affinity, lightly polymerized Abs. Finally, although the affinity for Ag is associated with elevated levels of polymerization, analysis of naive Ig revealed that the degree of polymerization alone, not affinity, appears sufficient to prolong Ig half life. PMID- 20018611 TI - Mice with a selective impairment of IFN-gamma signaling in macrophage lineage cells demonstrate the critical role of IFN-gamma-activated macrophages for the control of protozoan parasitic infections in vivo. AB - IFN-gamma has long been recognized as a cytokine with potent and varied effects in the immune response. Although its effects on specific cell types have been well studied in vitro, its in vivo effects are less clearly understood because of its diverse actions on many different cell types. Although control of multiple protozoan parasites is thought to depend critically on the direct action of IFN gamma on macrophages, this premise has never been directly proven in vivo. To more directly examine the effects of IFN-gamma on cells of the macrophage lineage in vivo, we generated mice called the "macrophages insensitive to IFN-gamma" (MIIG) mice, which express a dominant negative mutant IFN-gamma receptor in CD68+ cells: monocytes, macrophages, dendritic cells, and mast cells. Macrophage lineage cells and mast cells from these mice are unable to respond to IFN-gamma, whereas other cells are able to produce and respond to this cytokine normally. When challenged in vitro, macrophages from MIIG mice were unable produce NO or kill Trypanosoma cruzi or Leishmania major after priming with IFN-gamma. Furthermore, MIIG mice demonstrated impaired parasite control and heightened mortality after T. cruzi, L. major, and Toxoplasma gondii infection, despite an appropriate IFN-gamma response. In contrast, MIIG mice displayed normal control of lymphocytic choriomeningitis virus, despite persistent insensitivity of macrophages to IFN-gamma. Thus, the MIIG mouse formally demonstrates for the first time in vivo, the specific importance of direct, IFN-gamma mediated activation of macrophages for controlling infection with multiple protozoan parasites. PMID- 20018612 TI - Subversion of innate immune responses by Brucella through the targeted degradation of the TLR signaling adapter, MAL. AB - Gram-negative bacteria belonging to the Brucella species cause chronic infections that can result in undulant fever, arthritis, and osteomyelitis in humans. Remarkably, Brucella sp. genomes encode a protein, named TcpB, that bears significant homology with mammalian Toll/IL-1 receptor domains and whose expression causes degradation of the phosphorylated, signal competent form of the adapter MyD88-adapter-like (MAL). This effect of TcpB is mediated through its box 1 region and has no effect on other TLR adapter proteins such as MyD88 or TIR domain containing adapter protein-inducing IFNbeta. TcpB also does not affect a mutant, signal-incompetent form of MAL that cannot be phosphorylated. Interestingly, the presence of TcpB leads to enhanced polyubiquitination of MAL, which is likely responsible for its accelerated degradation. A Brucella abortus mutant lacking TcpB fails to reduce levels of MAL in infected macrophages. Therefore, TcpB represents a unique pathogen-derived molecule that suppresses host innate-immune responses by specifically targeting an individual adapter molecule in the TLR signaling pathway for degradation. PMID- 20018613 TI - Peroxiredoxin 1 stimulates secretion of proinflammatory cytokines by binding to TLR4. AB - Peroxiredoxin 1 (Prx1) is an antioxidant and molecular chaperone that can be secreted from tumor cells. Prx1 is overexpressed in many cancers, and elevation of Prx1 is associated with poor clinical outcome. In the current study, we demonstrate that incubation of Prx1 with thioglycollate-elicited murine macrophages or immature bone marrow-derived dendritic cells resulted in TLR4 dependent secretion of TNF-alpha and IL-6 and dendritic cell maturation. Optimal secretion of cytokines in response to Prx1 was dependent upon serum and required CD14 and MD2. Binding of Prx1 to thioglycollate macrophages occurred within minutes and resulted in TLR4 endocytosis. Prx1 interaction with TLR4 was independent of its peroxidase activity and appeared to be dependent on its chaperone activity and ability to form decamers. Cytokine expression occurred via the TLR-MyD88 signaling pathway, which resulted in nuclear translocation and activation of NF-kappaB. These findings suggest that Prx1 may act as danger signal similar to other TLR4-binding chaperone molecules such as HSP72. PMID- 20018614 TI - Fos proteins suppress dextran sulfate sodium-induced colitis through inhibition of NF-kappaB. AB - The Fos family proteins, c-Fos and Fra-1, are components of the dimeric transcription factor AP-1, which is typically composed of Fos and Jun family proteins. We have previously shown that mice lacking c-Fos (Fos(-/-) mice) respond more strongly to LPS injection than do wild-type (wt) controls. We then examined the sensitivity of Fos(-/-) mice to acute inflammatory stress in a dextran sulfate sodium (DSS)-induced colitis model. We found that Fos(-/-) mice exhibited more severe weight loss, bleeding, diarrhea, and colon shortening than did wt mice, in association with higher TNF-alpha production and NF-kappaB activity in colon segments of DSS-treated Fos(-/-) mice. Furthermore, NF-kappaB inhibition suppressed severe DSS-induced colitis in Fos(-/-) mice. In contrast, Fra-1 transgenic (Tg) mice responded poorly to LPS injection, and Fra-1 overexpressing macrophages and fibroblasts showed reduced production of proinflammatory cytokines, NO, and NF-kappaB activity. Remarkably, in the DSS induced colitis model, Fra-1 Tg mice showed less severe clinical scores of colitis than did wt mice. Consistently, proinflammatory cytokine production and NF-kappaB activity in colon segments of DSS-treated Fra-1 Tg mice were lower than in wt controls. These findings reveal that the absence of c-Fos and overexpression of Fra-1 respectively enhance and suppress the activation of NF kappaB in DSS-induced inflammatory stress. In this paper, we propose that AP-1 transcription factors containing c-Fos or Fra-1 are negative regulators of NF kappaB-mediated stress responses. PMID- 20018615 TI - RNA interference screen in primary human T cells reveals FLT3 as a modulator of IL-10 levels. AB - Functional studies of human primary immune cells have been hampered by the lack of tools to silence gene functions. In this study, we report the application of a lentiviral RNA interference library in primary human T cells. Using a subgenomic short hair RNA library targeting approximately 1000 signaling genes, we identified novel genes that control the levels of IL-10 produced. IL-10 is a potent anti-inflammatory cytokine secreted by several cell types, including T regulatory type 1 cells, a subset of T regulatory cells that exert their suppressive activity through IL-10 secretion. FLT3, a known hematopoeitic growth factor, was found to be a negative regulator of IL-10 levels in activated T cells. This was based on several observations. First, FLT3 and its ligand (FL) were both induced by T cell activation. Second, silencing of FLT3 led to increased IL-10 levels, whereas addition of FL suppressed IL-10 secretion and increased FLT3 surface levels. Third, engagement of CD46, a known inducer of T regulatory type 1 cells, upregulated surface FLT3, and secreted FL, which then inhibited IL-10 production by T cells. Hence, FL and FLT3 form a novel regulatory feedback loop that limits IL-10 production in T cells. Our results identified FLT3 as a new regulator of T cell function and offer a strategy to genetically dissect specific pathways in T cells. PMID- 20018616 TI - Granzyme A- and B-cluster deficiency delays acute lung injury in pneumovirus infected mice. AB - Lower respiratory tract infection by the human pneumovirus respiratory syncytial virus is a frequent cause of acute lung injury in children. Severe pneumovirus disease in humans is associated with activation of the granzyme pathway by effector lymphocytes, which may promote pathology by exaggerating proapoptotic caspase activity and proinflammatory activity. The main goal of this study was to determine whether granzymes contribute to the development of acute lung injury in pneumovirus-infected mice. Granzyme-expressing mice and granzyme A- and B-cluster single- and double-knockout mice were inoculated with the rodent pneumovirus pneumonia virus of mice strain J3666, and were studied for markers of lung inflammation and injury. Expression of granzyme A and B is detected in effector lymphocytes in mouse lungs in response to pneumovirus infection. Mice deficient for granzyme A and the granzyme B cluster have unchanged virus titers in the lungs but show a significantly delayed clinical response to fatal pneumovirus infection, a feature that is associated with delayed neutrophil recruitment, diminished activation of caspase-3, and reduced lung permeability. We conclude that granzyme A- and B-cluster deficiency delays the acute progression of pneumovirus disease by reducing alveolar injury. PMID- 20018617 TI - Cutting edge: Sympathetic nervous system increases proinflammatory cytokines and exacerbates influenza A virus pathogenesis. AB - Although the sympathetic nervous system innervates the lung, little is known about its participation in host immunity to pulmonary pathogens. In this study, we show that peripheral sympathectomy reduces mouse morbidity and mortality from influenza A virus-induced pneumonia due to reduced inflammatory influx of monocytes, neutrophils, and NK cells. Mortality was also delayed by treating mice with an alpha-adrenergic antagonist. Sympathectomy diminished the immediate innate cytokine responses, particularly IL-1, which was profoundly reduced. These findings demonstrate an unexpected role for the sympathetic nervous system in innate antiviral immunity and in exacerbating the pathology of a virus of great significance to human and animal health. PMID- 20018618 TI - Signaling role for lysophosphatidylcholine acyltransferase 3 in receptor regulated arachidonic acid reacylation reactions in human monocytes. AB - Cellular availability of free arachidonic acid (AA) is an important step in the production of pro- and anti-inflammatory eicosanoids. Control of free AA levels in cells is carried out by the action of phospholipase A2s and lysophospholipid acyltransferases, which are responsible for the reactions of deacylation and incorporation of AA from and into the sn-2 position of phospholipids, respectively. In this work, we have examined the pathways for AA incorporation into phospholipids in human monocytes stimulated by zymosan. Our data show that stimulated cells exhibit an enhanced incorporation of AA into phospholipids that is not secondary to an increased availability of lysophospholipid acceptors due to phospholipase A2 activation but rather reflects the receptor-regulated nature of the AA reacylation pathway. In vitro activity measurements indicate that the receptor-sensitive step of the AA reacylation pathway is the acyltransferase using lysophosphatidylcholine (lysoPC) as acceptor, and inhibition of the enzyme lysoPC acyltransferase 3 by specific small interfering RNA results in inhibition of the stimulated incorporation of AA into phospholipids. Collectively, these results define lysoPC acyltransferase 3 as a novel-signal-regulated enzyme that is centrally implicated in limiting free AA levels in activated cells. PMID- 20018619 TI - Independent regulation of chemokine responsiveness and cytolytic function versus CD8+ T cell expansion by dendritic cells. AB - The ability of cancer vaccines to induce tumor-specific CD8+ T cells in the circulation of cancer patients has been shown to poorly correlate with their clinical effectiveness. In this study, we report that although Ags presented by different types of mature dendritic cells (DCs) are similarly effective in inducing CD8+ T cell expansion, the acquisition of CTL function and peripheral type chemokine receptors, CCR5 and CXCR3, requires Ag presentation by a select type of DCs. Both "standard" DCs (matured in the presence of PGE2) and type 1 polarized DCs (DC1s) (matured in the presence of IFNs and TLR ligands, which prevent DCs "exhaustion") are similarly effective in inducing CD8+ T cell expansion and acquisition of CD45RO+IL-7R+IL-15R+ phenotype. However, granzyme B expression, acquisition of CTL activity, and peripheral tissue-type chemokine responsiveness are features exclusively exhibited by CD8+ T cells activated by DC1s. This advantage of DC1s was observed in polyclonally activated naive and memory CD8(+) T cells and in blood-isolated melanoma-specific CTL precursors. Our data help to explain the dissociation between the ability of cancer vaccines to induce high numbers of tumor-specific CD8+ T cells in the blood of cancer patients and their ability to promote clinical responses, providing for new strategies of cancer immunotherapy. PMID- 20018620 TI - Gene expression analysis of macrophages that facilitate tumor invasion supports a role for Wnt-signaling in mediating their activity in primary mammary tumors. AB - The tumor microenvironment modifies the malignancy of tumors. In solid tumors, this environment is populated by many macrophages that, in genetic studies that depleted these cells from mouse models of breast cancer, were shown to promote tumor progression to malignancy and increase metastatic potential. Mechanistic studies showed that these tumor-promoting effects of macrophages are through the stimulation of tumor cell migration, invasion, intravasation, and enhancement of angiogenesis. Using an in vivo invasion assay, it was demonstrated that invasive carcinoma cells are a unique subpopulation of tumor cells whose invasion and chemotaxis is dependent on the comigration of tumor-associated macrophages (TAMs) with obligate reciprocal signaling through an epidermal growth factor-CSF-1 paracrine loop. In this study, these invasion-promoting macrophages were isolated and subjected to analysis of their transcriptome in comparison with TAMs isolated indiscriminately to function using established macrophage markers. Unsupervised analysis of transcript patterns showed that the invasion-associated TAMs represent a unique subpopulation of TAMs that, by gene ontology criteria, have gene expression patterns related to tissue and organ development. Gene set enrichment analysis showed that these macrophages are also specifically enriched for molecules involved in Wnt-signaling. Previously, it was shown that macrophage derived Wnt molecules promote vascular remodeling and that tumor cells are highly motile and intravasate around perivascular TAM clusters. Taken together, we conjecture that invasive TAMs link angiogenesis and tumor invasion and that Wnt signaling plays a role in mediating their activity. PMID- 20018621 TI - Terminal deoxynucleotidyl transferase is required for an optimal response to the polysaccharide alpha-1,3 dextran. AB - An understanding of Ab responses to polysaccharides associated with pathogenic microorganisms is of importance for improving vaccine design, especially in neonates that respond poorly to these types of Ags. In this study, we have investigated the role of the lymphoid-specific enzyme TdT in generating B cell clones responsive to alpha-1,3 dextran (DEX). TdT is a DNA polymerase that plays a major role in generating diversity of lymphocyte AgRs during V(D)J recombination. In this study, we show that the DEX-specific Ab response is lower, and the dominant DEX-specific J558 idiotype (Id) is not detected in TdT(-/-) mice when compared with wild-type (WT) BALB/c mice. Nucleotide sequencing of H chain CDR3s of DEX-specific plasmablasts, sorted postimmunization, showed that TdT(-/-) mice generate a lower frequency of the predominant adult molecularly determined clone J558. Complementation of TdT expression in TdT(-/-) mice by early forced expression of the short splice variant of TdT-restored WT proportions of J558 Id+ clones and also abrogated the development of the minor M104E Id+ clones. J558 Id V(D)J rearrangements are detected as early as 7 d after birth in IgM-negative B cell precursors in the liver and spleen of WT and TdT-transgenic mice but not in TdT(-/-) mice. These data show that TdT is essential for the generation of the predominant higher-affinity DEX-responsive J558 clone. PMID- 20018623 TI - Heat shock transcription factor 1 inhibits expression of IL-6 through activating transcription factor 3. AB - The febrile response is a complex physiological reaction to disease, including a cytokine-mediated increase in body temperature and the activation of inflammatory systems. Fever has beneficial roles in terms of disease prognosis, partly by suppressing the expression of inflammatory cytokines. However, the molecular mechanisms underlining the fever-mediated suppression of inflammatory gene expression have not been clarified. In this study, we showed that heat shock suppresses LPS-induced expression of IL-6, a major pyrogenic cytokine, in mouse embryonic fibroblasts and macrophages. Heat shock transcription factor 1 (HSF1) activated by heat shock induced the expression of activating transcription factor (ATF) 3, a negative regulator of IL-6, and ATF3 was necessary for heat-mediated suppression of IL-6, indicating a fever-mediated feedback loop consisting of HSF1 and ATF3. A comprehensive analysis of inflammatory gene expression revealed that heat pretreatment suppresses LPS-induced expression of most genes (86%), in part (67%) via ATF3. When HSF1-null and ATF3-null mice were injected with LPS, they expressed much higher levels of IL-6 than wild-type mice, resulting in an exaggerated febrile response. These results demonstrate a novel inhibitory pathway for inflammatory cytokines. PMID- 20018622 TI - Differential regulation of IL-4Ralpha expression by antigen versus cytokine stimulation characterizes Th2 progression in vivo. AB - IL-4 promotes Th2 differentiation and provides immunity to helminth infections but is also associated with allergy and asthma. This suggests that precise adjustment of IL-4 responsiveness is needed to correctly balance immune responses. The IL-4Ralpha chain is an essential component of the IL-4 receptor and signals via STAT6. In this study, we show that infection with a helminth pathogen elicited broad upregulation of IL-4Ralpha on bystander CD4+ T cells in the draining lymph node, while simultaneously resulting in the loss of IL-4Ralpha expression on activated Th2 cells. IL-4Ralpha upregulation was restricted to the reactive lymph node, occurred within 4 d of infection, and was driven by an IL-4- and STAT6-dependent mechanism. Mice heterozygous for Stat6 exhibited reduced IL 4Ralpha upregulation and a correspondingly attenuated Th2 response. Indeed, the enhanced IL-4Ralpha upregulation in BALB/c mice, compared with that in C57BL6 mice, predicted their stronger Th2 response. The selective downregulation of IL 4Ralpha on highly activated Th cells was triggered by antigenic stimulation, was accompanied by loss of IL-7Ralpha, and rendered the cells unresponsive to IL-4. Together these data reveal a tightly controlled program of changing IL-4 responsiveness that characterizes the initiation, amplification, and restriction of a Th2 response in vivo. PMID- 20018624 TI - Hypochlorous acid: a natural adjuvant that facilitates antigen processing, cross priming, and the induction of adaptive immunity. AB - The production of hypochlorous acid (HOCl) is a characteristic of granulocyte activation, a hallmark of the early phase of innate immune responses. In this study, we show that, in addition to its well-established role as a microbicide, HOCl can act as a natural adjuvant of adaptive immunity. HOCl enhances the T cell responses to the model Ag OVA, facilitating the processing and presentation of this protein via the class II MHC pathway. HOCl modification also enhances cross presentation of the tumor Ag tyrosinase-related protein 2 via class I MHC. The adjuvant effects of HOCl are independent of TLR signaling. The enhanced presentation of HOCl-modified OVA is mediated via modification of the N-linked carbohydrate side chain rather than formation of protein aldehydes or chloramines. HOCl-modified OVA is taken up more efficiently by APCs and is degraded more efficiently by proteinases. Atomic force microscopy demonstrated that enhanced uptake is mediated via specific receptor binding, one candidate for which is the scavenger receptor lectin-like oxidized low-density lipoprotein receptor, which shows enhanced binding to chlorinated OVA. A function of HOCl is therefore to target glycoprotein Ags to scavenger receptors on the APC surface. This additional mechanism linking innate and adaptive immunity suggests novel strategies to enhance immunity to vaccines. PMID- 20018626 TI - Cytohesin-1 regulates the Arf6-phospholipase D signaling axis in human neutrophils: impact on superoxide anion production and secretion. AB - Polymorphonuclear neutrophil (PMN) stimulation with fMLP stimulates small G proteins such as ADP-ribosylation factors (Arfs) Arf1 and Arf6, leading to phospholipase D (PLD) activation and functions such as degranulation and the oxidative burst. However, the molecular links between fMLF receptors and PLD remain unclear. PMNs express cytohesin-1, an Arf-guanine exchange factor that activates Arfs, and its expression is strongly induced during the acquisition of the neutrophilic phenotype by neutrophil-like cells. The role of cytohesin-1 in the activation of the fMLF-Arf-PLD signaling axis, and the accomplishment of superoxide anion production, and degranulation was investigated in PMNs using the selective inhibitor of cytohesin, Sec 7 inhibitor H3 (secinH3). Cytohesin-1 inhibition with secinH3 leads to Arf6 but not Arf1 inhibition, demonstrating the specificity for Arf6, and fMLF-mediated activation of PLD and of the oxidative burst as well. We observed a decrease in fMLF-mediated protein secretion and expression of cell surface markers corresponding to primary (CD63/myeloperoxidase), secondary (CD66/lactoferrin), and tertiary (matrix metalloproteinase-9) granules in PMNs incubated with secinH3. Similarly, silencing cytohesin-1 or Arf6 in PLB-985 cells negatively affected fMLF-induced activation of PLD, superoxide production, and expression of granule markers on the cell surface. In contrast, stable overexpression of cytohesin-1 in PLB-985 cells enhanced fMLF-induced activation of Arf6, PLD, and NADPH oxidase. The results of this study provide evidence for an involvement of cytohesin-1 in the regulation of the functional responses of human PMNs and link these events, in part at least, to the activation of Arf6. PMID- 20018625 TI - Neurons preferentially respond to self-MHC class I allele products regardless of peptide presented. AB - Studies of mice lacking MHC class I (MHC I)-associated proteins have demonstrated a role for MHC I in neurodevelopment. A central question arising from these observations is whether neuronal recognition of MHC I has specificity for the MHC I allele product and the peptide presented. Using a well-established embryonic retina explant system, we observed that picomolar levels of a recombinant self MHC I molecule inhibited neurite outgrowth. We then assessed the neurobiological activity of a panel of recombinant soluble MHC Is, consisting of different MHC I heavy chains with a defined self- or nonself-peptide presented, on cultured embryonic retinas from mice with different MHC I haplotypes. We observed that self-MHC I allele products had greater inhibitory neuroactivity than nonself-MHC I molecules, regardless of the nature of the peptide presented, a pattern akin to MHC I recognition by some innate immune system receptors. However, self-MHC I molecules had no effect on retinas from MHC I-deficient mice. These observations suggest that neuronal recognition of MHC I may be coordinated with the inherited MHC I alleles, as occurs in the innate immune system. Consistent with this notion, we show that MHC I and MHC I receptors are coexpressed by precursor cells at the earliest stages of retina development, which could enable such coordination. PMID- 20018627 TI - Cutting edge: lymphatic vessels, not blood vessels, primarily mediate immune rejections after transplantation. AB - The purpose of this study was to determine the relative importance of blood vessels (hemangiogenesis) versus lymphatic vessels (lymphangiogenesis) in mediating immunological responses after transplantation. Using the murine model of corneal transplantation, graft survival was compared in differentially prevascularized and avascular recipient beds. Donor corneas (C57BL/6) were transplanted into uninflamed or inflamed avascular, prehemvascularized only or prehemvascularized and prelymphvascularized recipient murine eyes (BALB/C). Selective inhibition of lymphangiogenesis was achieved using antivascular endothelial growth factor receptor 3 Abs and anti-integrin alpha5 small molecules. Grafts placed into only prehemvascularized recipient beds had a similarly good graft survival compared with grafts placed into completely avascular, normal recipients, whereas the pre-existence of lymphatic vessels significantly deteriorated corneal graft survival (p < 0.05). Lymphatic vessels seem to contribute significantly to graft rejection after (corneal) transplantation. That may allow for selective, temporary, perioperative antilymphangiogenic treatment to promote graft survival without affecting blood vessels, even after solid organ transplantation. PMID- 20018628 TI - PDCA expression by B lymphocytes reveals important functional attributes. AB - We have demonstrated in this study the existence of a PDCA-expressing functional B cell population (PDCA+ B lymphocytes), which differentiates from activated conventional B (PDCA-IgM+) lymphocytes. Stimulation with anti-micro, LPS, CpG oligodeoxynucleotide, HSV-1, or CTLA-4 Ig activates the PDCA+ B lymphocytes, leading to cell division and induction of type I IFNs and IDO. Notably, the PDCA+ B lymphocytes are capable of Ag-specific Ab production and Ig class switching, which is corroborated by transfer experiments in B- and PDCA+ B lymphocyte deficient microMT mice. Importantly, in lupus-prone MRL-Fas(lpr) mice, PDCA+ B lymphocytes remain the principal source of autoantibodies. The PDCA+ B lymphocytes have phenotypes with plasmacytoid dendritic cells, but are a distinct cell population in that they develop from C-kit+B220+ pro-B precursors. Thus, our data suggest that not all PDCA+ cells are dendritic cell-derived plasmacytoid dendritic cells and that a significant majority is the PDCA+ B lymphocyte population having distinct phenotype and function. PMID- 20018629 TI - Presenilin/gamma-secretase cleaves CD46 in response to Neisseria infection. AB - CD46 is a type I transmembrane protein with complement and T cell regulatory functions in human cells. CD46 has signaling and receptor properties in immune and nonimmune cells, many of which are dependent on the expression of cytoplasmic tail (cyt) isoforms cyt1 or cyt2. Little is known about how cyt1 and cyt2 mediate cellular responses. We show that CD46-cyt1 and CD46-cyt2 are substrates for presenilin/gamma-secretase (PS/gammaS), an endogenous protease complex that regulates many important signaling proteins through proteolytic processing. PS/gammaS processing of CD46 releases immunoprecipitable cyt1 and cyt2 tail peptides into the cell, is blocked by chemical inhibitors, and is prevented in dominant negative presenilin mutant cell lines. Two human pathogens, Neisseria gonorrhoeae and Neisseria meningitidis, stimulate PS/gammaS processing of CD46 cyt1 and CD46-cyt2. This stimulation requires type IV pili and PilT, the type IV pilus retraction motor, implying that mechanotransduction plays a role in this event. We present a model for PS/gammaS processing of CD46 that provides a mechanism by which signals are transduced via the cyt1 and cyt2 tails to regulate CD46-dependent cellular responses. Our findings have broad implications for understanding the full range of CD46 functions in infection and noninfection situations. PMID- 20018630 TI - AIDS progression is associated with the emergence of IL-17-producing cells early after simian immunodeficiency virus infection. AB - IL-17 is a potent effector cytokine involved in inflammatory response and antimicrobial defense. We report that SIV infection of rhesus macaques (RMs) results in the emergence of IL-17-expressing cells during the acute phase. This subpopulation appears at day 14 postinfection concomitantly with an increase in TGF-beta and IL-18 expression. This subset, which exhibits phenotypic markers of NK T cells (NKT), rather than Th17 CD4 cells, persists during the chronic phase and is higher in noncontrollers SIV-infected RMs compared with controllers SIV infected RMs. In contrast, in the nonpathogenic model of SIVagm infection of African green monkeys, no change in the level of IL-17-expressing cells is observed in lymphoid organs. Consistent with the emergence of TGF-beta and IL-18 during the acute phase in SIV-infected RMs, but not in SIV-infected African green monkeys, we demonstrate that in vitro TGF-beta and IL-18 induce the differentiation and expansion of IL-17+NKT+. Altogether, these results demonstrate that IL-17-producing NKT are associated with the pathogenesis of SIV in RMs and suggest that TGF-beta and IL-18 play a role in their development. PMID- 20018632 TI - Prostaglandin E2 inhibits IFN-alpha secretion and Th1 costimulation by human plasmacytoid dendritic cells via E-prostanoid 2 and E-prostanoid 4 receptor engagement. AB - Plasmacytoid dendritic cell (PDC)-derived IFN-alpha plays a central role in antiviral defense and in Th1-driven autoimmune diseases, such as systemic lupus erythematosus (SLE). In the current study, we explored how PGE2 effects the phenotype of PDCs from healthy and SLE subjects. Although PGE2 is considered to mediate mainly proinflammatory effects, we show that PGE2 and PG analogs potently inhibit secretion of IFN-alpha by TLR-activated PDCs. This effect is mainly mediated by PG receptors E-prostanoid 2 and E-prostanoid 4 and involves inhibition of IFN regulatory factor 7 expression. Of note, profound IFN-alpha inhibition by PGE2 is also seen in PDCs from SLE subjects, independent of age, disease activity, and therapy. We show that TLR9-activated PDCs treated with PGE2 exhibit DC2-like characteristics with enhanced expression of CD86 and CD62L, and decreased expression of CD80 and MHC class I. Consequently, PGE2-treated PDCs suppress secretion of Th1 cytokines by T cells while increasing the secretion of Th2 cytokines. Prevention of CpG-induced CD62L downregulation by PGE2 suggests that it may induce the retreat of PDCs from inflamed tissues. Our data on the effects of PGE2 on PDCs may explain occasional reports about the induction of SLE like symptoms by cyclooxygenase inhibitors as well as improvement of such symptoms by treatment with PG analogs. In conclusion, our data suggest that PGE2 and certain PG analogs, some of which are already in clinical use, should be evaluated as a novel and inexpensive treatment approach for patients with SLE and other IFN-alpha-dependent, Th1-driven autoimmune diseases. PMID- 20018633 TI - Matrix architecture dictates three-dimensional migration modes of human macrophages: differential involvement of proteases and podosome-like structures. AB - Tissue infiltration of macrophages, although critical for innate immunity, is also involved in pathologies, such as chronic inflammation and cancer. In vivo, macrophages migrate mostly in a constrained three-dimensional (3D) environment. However, in vitro studies, mainly focused on two dimensions, do not provide meaningful clues about the mechanisms involved in 3D macrophage migration. In contrast, tumor cell 3D migration is well documented. It comprises a protease independent and Rho kinase (ROCK)-dependent amoeboid migration mode and a protease-dependent and ROCK-independent mesenchymal migration mode. In this study, we examined the influence of extracellular matrix (composition, architecture, and stiffness) on 3D migration of human macrophages derived from blood monocytes (MDMs). We show that: 1) MDMs use either the amoeboid migration mode in fibrillar collagen I or the mesenchymal migration mode in Matrigel and gelled collagen I, whereas HT1080 tumor cells only perform mesenchymal migration; 2) when MDMs use the mesenchymal migratory mode, they form 3D collagenolytic structures at the tips of cell protrusions that share several markers with podosomes as described in two dimensions; 3) in contrast to tumor cells, matrix metalloproteinase inhibitors do not impair protease-dependent macrophage 3D migration, suggesting the involvement of other proteolytic systems; and 4) MDMs infiltrating matrices of similar composition but with variable stiffness adapt their migration mode primarily to the matrix architecture. In conclusion, although it is admitted that leukocytes 3D migration is restricted to the amoeboid mode, we show that human macrophages also perform the mesenchymal mode but in a distinct manner than tumor cells, and they naturally adapt their migration mode to the environmental constraints. PMID- 20018634 TI - Teaching NeuroImages: Molar tooth sign with hypotonia, ataxia, and nystagmus (Joubert syndrome) and hypothyroidism. PMID- 20018631 TI - Development of murine lupus involves the combined genetic contribution of the SLAM and FcgammaR intervals within the Nba2 autoimmune susceptibility locus. AB - Autoantibodies are of central importance in the pathogenesis of Ab-mediated autoimmune disorders. The murine lupus susceptibility locus Nba2 on chromosome 1 and the syntenic human locus are associated with a loss of immune tolerance that leads to antinuclear Ab production. To identify gene intervals within Nba2 that control the development of autoantibody-producing B cells and to determine the cellular components through which Nba2 genes accomplish this, we generated congenic mice expressing various Nba2 intervals where genes for the FcgammaR, SLAM, and IFN-inducible families are encoded. Analysis of congenic strains demonstrated that the FcgammaR and SLAM intervals independently controlled the severity of autoantibody production and renal disease, yet are both required for lupus susceptibility. Deregulated homeostasis of terminally differentiated B cells was found to be controlled by the FcgammaR interval where FcgammaRIIb mediated apoptosis of germinal center B cells and plasma cells was impaired. Increased numbers of activated plasmacytoid dendritic cells that were distinctly CD19+ and promoted plasma cell differentiation via the proinflammatory cytokines IL-10 and IFNalpha were linked to the SLAM interval. These findings suggest that SLAM and FcgammaR intervals act cooperatively to influence the clinical course of disease through supporting the differentiation and survival of autoantibody producing cells. PMID- 20018636 TI - Differential association of [11C]PIB and [18F]FDDNP binding with cognitive impairment. AB - OBJECTIVE: To evaluate associations of [(11)C]Pittsburgh compound B (PIB) and [(18)F]FDDNP with impairment in specific cognitive domains over the broader spectrum comprising cognitively normal elderly subjects, patients with mild cognitive impairment (MCI), and patients with Alzheimer disease (AD). METHODS: Twelve patients with AD, 13 patients with MCI, and 15 cognitively normal elderly subjects were included. Paired [(11)C]PIB and [(18)F]FDDNP PET scans were performed in all subjects. Binding potential (BP(ND)) was calculated using parametric images of BP(ND) for global, frontal, parietal, and temporal cortex; medial temporal lobe; and posterior cingulate. Cognitive functions were assessed using a battery of neuropsychological tests. Linear regression analyses were used to assess associations of [(11)C]PIB and [(18)F]FDDNP binding with cognitive measures. RESULTS: Adjusted for age, sex, and [(18)F]FDDNP binding, higher global [(11)C]PIB binding was associated with lower scores on the Mini-Mental State Examination, immediate and delayed recall of the Rey Auditory Verbal Learning Task (RAVLT), Visual Association Task, and Trail Making Test part B. Conversely, higher [(18)F]FDDNP binding was independently associated with lower scores on immediate recall of the RAVLT. After additional adjustment for diagnosis, higher [(11)C]PIB binding remained independently associated with delayed recall (standardized beta = -0.39, p = 0.01), whereas higher [(18)F]FDDNP binding remained independently associated with immediate recall (standardized beta = 0.32, p = 0.03). When regional binding was assessed using stepwise models, both increased frontal [(11)C]PIB and temporal [(18)F]FDDNP binding were associated with memory, whereas increased parietal [(11)C]PIB binding was associated with nonmemory functions. CONCLUSION: Increased [(18)F]FDDNP binding is specifically associated with impairment of episodic memory, whereas increased [(11)C]Pittsburgh compound B binding is associated with impairment in a broader range of cognitive functions. PMID- 20018637 TI - Medical decision-making capacity in patients with malignant glioma. AB - OBJECTIVE: Patients with malignant glioma (MG) must make ongoing medical treatment decisions concerning a progressive disease that erodes cognition. We prospectively assessed medical decision-making capacity (MDC) in patients with MG using a standardized psychometric instrument. METHODS: Participants were 22 healthy controls and 26 patients with histologically verified MG. Group performance was compared on the Capacity to Consent to Treatment Instrument (CCTI), a psychometric measure of MDC incorporating 4 standards (choice, understanding, reasoning, and appreciation), and on neuropsychological and demographic variables. Capacity outcomes (capable, marginally capable, or incapable) on the CCTI standards were identified for the MG group. Within the MG group, scores on demographic, clinical, and neuropsychological variables were correlated with scores on each CCTI standard, and significant bivariate correlates were subsequently entered into exploratory stepwise regression analyses to identify multivariate cognitive predictors of the CCTI standards. RESULTS: Patients with MG performed significantly below controls on consent standards of understanding and reasoning, and showed a trend on appreciation. Relative to controls, more than 50% of the patients with MG demonstrated capacity compromise (marginally capable or incapable outcomes) in MDC. In the MG group, cognitive measures of verbal acquisition/recall and, to a lesser extent, semantic fluency predicted performance on the appreciation, reasoning, and understanding standards. Karnofsky score was also associated with CCTI performance. CONCLUSIONS: Soon after diagnosis, patients with malignant glioma (MG) have impaired capacity to make treatment decisions relative to controls. Medical decision-making capacity (MDC) impairment in MG seems to be primarily related to the effects of short-term verbal memory deficits. Ongoing assessment of MDC in patients with MG is strongly recommended. PMID- 20018638 TI - HLA-DRB1 and month of birth in multiple sclerosis. AB - BACKGROUND: Multiple sclerosis (MS) displays a month-of-birth effect, with an excess of individuals being born in the spring and a deficit in the winter. This effect was shown to be more pronounced in familial cases of MS. In the present study, we investigated whether this month-of-birth association has any relation to the principal MS susceptibility gene, HLA-DRB1. METHODS: A total of 4,834 patients with MS, 4,056 controls, and 659 unaffected siblings from Canada, Sweden, and Norway were genotyped for the HLA-DRB1 gene. Month of birth was compared for patients, controls, and unaffected siblings with and without the MS risk allele HLA-DRB1*15. RESULTS: Significantly fewer patients with MS carrying the HLA-DRB1*15 risk allele were born in November compared with patients not carrying this allele (p = 0.02). Additionally, patients with MS carrying HLA DRB1*15 had a higher number of April births compared with patients with MS not carrying HLA-DRB1*15 (p = 0.004). These differences were not present in controls or unaffected siblings. CONCLUSIONS: Month of birth, HLA-DRB1 genotype, and risk of multiple sclerosis are associated. The interaction of a seasonal risk factor with loci at or near HLA-DRB1 during gestation or shortly after birth is implicated. PMID- 20018640 TI - Characteristic brain MRI appearance of Erdheim-Chester disease. PMID- 20018639 TI - Road safety in drivers with Parkinson disease. AB - OBJECTIVE: To assess road safety and its predictors in drivers with Parkinson disease (PD). METHODS: Licensed, active drivers with PD (n = 84; age = 67.3 +/- 7.8, median Hoehn & Yahr stage II) and controls (n = 182; age = 67.6 +/- 7.5) underwent cognitive, visual, and motor tests, and drove a standardized route in urban and rural settings in an instrumented vehicle. Safety errors were judged and documented by a driving expert based on video data review. RESULTS: Drivers with PD committed more total safety errors compared to controls (41.6 +/- 14.6 vs 32.9 +/- 12.3, p < 0.0001); 77.4% of drivers with PD committed more errors than the median total error count of the controls (medians: PD = 39.5, controls = 31.0). Lane violations were the most common error category in both groups. Group differences in some error categories became insignificant after results were adjusted for demographics and familiarity with the local driving environment. The PD group performed worse on tests of motor, cognitive, and visual abilities. Within the PD group, older age and worse performances on tests of visual acuity, contrast sensitivity, attention, visuospatial abilities, visual memory, and general cognition predicted error counts. Measures of visual processing speed and attention and far visual acuity were jointly predictive of error counts in a multivariate model. CONCLUSIONS: Overall, drivers with Parkinson disease (PD) had poorer road safety compared to controls, but there was considerable variability among the drivers with PD, and some performed normally. Familiarity with the driving environment was a mitigating factor against unsafe driving in PD. Impairments in visual perception and cognition were associated with road safety errors in drivers with PD. PMID- 20018641 TI - Evolution of oculomotor and clinical findings in autopsy-proven Richardson syndrome. PMID- 20018642 TI - Novel compound heterozygous ALS2 mutations cause juvenile amyotrophic lateral sclerosis in Japan. PMID- 20018643 TI - Intergenerational contraction of the CTG repeats in 2 families with myotonic dystrophy type 1. PMID- 20018644 TI - Congenital null mutations of the FOLR1 gene: a progressive neurologic disease and its treatment. PMID- 20018645 TI - Stimulation of the parietal cortex affects reaching in a patient with epilepsy. PMID- 20018646 TI - Large choroid plexus teratoma: A rare cause of congenital hydrocephalus. PMID- 20018647 TI - Stiff-person syndrome with amphiphysin antibodies: distinctive features of a rare disease. PMID- 20018648 TI - The essential neurologic examination: what should medical students be taught? PMID- 20018649 TI - Effect of intrinsic organic carbon on the optical properties of fresh diesel soot. AB - This study focuses on the retrieval of the normalized mass absorption cross section (MAC) of soot using theoretical calculations that incorporate new measurements of the optical properties of organic carbon (OC) intrinsic to fresh diesel soot. Intrinsic OC was extracted by water and an organic solvent, and the complex refractive index of the extracted OC was derived at 532 and 355-nm wavelengths using cavity ring-down aerosol spectrometry. The extracted OC was found to absorb weakly in the visible wavelengths and moderately at blue wavelengths. The mass ratio of OC and elemental carbon (EC) in the collected particles was evaluated using a thermo-optical method. The measured EC/OC ratio in the soot exhibited substantial variability from measurement to measurement, ranging between 2 and 5. To test the sensitivity of the MAC to this variability, three different EC/OC ratios (21, 11, and 12) were chosen as representative. Particle size and spherule morphology were estimated using scanning electron microscopy, and the soot was found to be primarily in the form of aggregates with a dominant aggregate diameter mode in the range 200-250 nm. The measured refractive index of the extracted OC was used with a variety of theoretical models to calculate the MAC of internally mixed diesel soot at 532 and 355 nm. We conclude that Rayleigh-Debye-Gans theory on clusters of coated spherules and T matrix of a solid EC spheroid coated by intrinsic OC are both consistent with previous measurements; however, Rayleigh-Debye-Gans theory provides a more realistic physical model for the calculation. PMID- 20018650 TI - Quantitative biological imaging by ptychographic x-ray diffraction microscopy. AB - Recent advances in coherent x-ray diffractive imaging have paved the way to reliable and quantitative imaging of noncompact specimens at the nanometer scale. Introduced a year ago, an advanced implementation of ptychographic coherent diffractive imaging has removed much of the previous limitations regarding sample preparation and illumination conditions. Here, we apply this recent approach toward structure determination at the nanoscale to biological microscopy. We show that the projected electron density of unstained and unsliced freeze-dried cells of the bacterium Deinococcus radiodurans can be derived from the reconstructed phase in a straightforward and reproducible way, with quantified and small errors. Thus, the approach may contribute in the future to the understanding of the highly disputed nucleoid structure of bacterial cells. In the present study, the estimated resolution for the cells was 85 nm (half-period length), whereas 50 nm resolution was demonstrated for lithographic test structures. With respect to the diameter of the pinhole used to illuminate the samples, a superresolution of about 15 was achieved for the cells and 30 for the test structures, respectively. These values should be assessed in view of the low dose applied on the order of approximately 1.3x10(5) Gy, and were shown to scale with photon fluence. PMID- 20018652 TI - Autocatalysis and organocatalysis with synthetic structures. AB - The discovery of ribozymes led to the proposal of an RNA world, where a single type of molecule was supposedly capable of self-replication and chemical catalysis. We show here that both autocatalysis and organocatalysis can be engineered into a synthetic structure. The compound is shown to selectively accelerate its own formation and catalyze either hydrogenation or nucleophilic addition to alpha,beta-unsaturated aldehydes. The observed reactivity indicates that the components of a purported pre-RNA world conceivably include smaller organic molecules. PMID- 20018651 TI - Beta-arrestin- but not G protein-mediated signaling by the "decoy" receptor CXCR7. AB - Ubiquitously expressed seven-transmembrane receptors (7TMRs) classically signal through heterotrimeric G proteins and are commonly referred to as G protein coupled receptors. It is now recognized that 7TMRs also signal through beta arrestins, which act as versatile adapters controlling receptor signaling, desensitization, and trafficking. Most endogenous receptors appear to signal in a balanced fashion using both beta-arrestin and G protein-mediated pathways. Some 7TMRs are thought to be nonsignaling "decoys" because of their inability to activate typical G protein signaling pathways; it has been proposed that these receptors act to scavenge ligands or function as coreceptors. Here we demonstrate that ligand binding to the decoy receptor CXCR7 does not result in activation of signaling pathways typical of G proteins but does activate MAP kinases through beta-arrestins in transiently transfected cells. Furthermore, we observe that vascular smooth muscle cells that endogenously express CXCR7 migrate to its ligand interferon-inducible T-cell alpha chemoattractant (ITAC), an effect that is significantly attenuated by treatment with either a CXCR7 antagonist or beta arrestin depletion by siRNA. This example of an endogenous "beta-arrestin-biased" 7TMR that signals through beta-arrestin in the absence of G protein activation demonstrates that some 7TMRs encoded in the genome have evolved to signal through beta-arrestin exclusively and suggests that other receptors that are currently thought to be orphans or decoys may also signal through such nonclassical pathways. PMID- 20018653 TI - Spontaneous formation of the unlocked state of the ribosome is a multistep process. AB - The mechanism of substrate translocation through the ribosome is central to the rapid and faithful translation of mRNA into proteins. The rate-limiting step in translocation is an unlocking process that includes the formation of an "unlocked" intermediate state, which requires the convergence of large-scale conformational events within the ribosome including tRNA hybrid states formation, closure of the ribosomal L1 stalk domain, and subunit ratcheting. Here, by imaging of the pretranslocation ribosome complex from multiple structural perspectives using two- and three-color single-molecule fluorescence resonance energy transfer, we observe that tRNA hybrid states formation and L1 stalk closure, events central to the unlocking mechanism, are not tightly coupled. These findings reveal that the unlocked state is achieved through a stochastic multistep process, where the extent of conformational coupling depends on the nature of tRNA substrates. These data suggest that cellular mechanisms affecting the coupling of conformational processes on the ribosome may regulate the process of translation elongation. PMID- 20018654 TI - Probiotics promote gut health through stimulation of epithelial innate immunity. AB - Probiotic formulations are widely available and have a variety of proposed beneficial effects, including promotion of gut health. The mechanisms of action of probiotic bacteria in the intestine are still unclear but are generally attributed to an antiinflammatory effect. Here, we demonstrate that the multiple probiotic formulation VSL#3 prevents the onset of intestinal inflammation by local stimulation of epithelial innate immune responses (i.e., increased production of epithelial-derived TNF-alpha and restoration of epithelial barrier function in vivo). We also demonstrate that probiotic bacteria stimulate epithelial production of TNF-alpha and activate NF-kappaB in vitro. Our results support the hypothesis that probiotics promote gut health through stimulation, rather than suppression, of the innate immune system. Furthermore, our findings provide the perspective that defects in innate immunity may play a critical role in the pathogenesis and progression of intestinal disorders, such as inflammatory bowel disease. PMID- 20018655 TI - Chloroplast acetyl-CoA carboxylase activity is 2-oxoglutarate-regulated by interaction of PII with the biotin carboxyl carrier subunit. AB - The PII protein is a signal integrator involved in the regulation of nitrogen metabolism in bacteria and plants. Upon sensing of cellular carbon and energy availability, PII conveys the signal by interacting with target proteins, thereby modulating their biological activity. Plant PII is located to plastids; therefore, to identify new PII target proteins, PII-affinity chromatography of soluble extracts from Arabidopsis leaf chloroplasts was performed. Several proteins were retained only when Mg-ATP was present in the binding medium and they were specifically released from the resin by application of a 2-oxoglutarate containing elution buffer. Mass spectroscopy of SDS/PAGE-resolved protein bands identified the biotin carboxyl carrier protein subunits of the plastidial acetyl CoA carboxylase (ACCase) and three other proteins containing a similar biotin/lipoyl-binding motif as putative PII targets. ACCase is a key enzyme initiating the synthesis of fatty acids in plastids. In in vitro reconstituted assays supplemented with exogenous ATP, recombinant Arabidopsis PII inhibited chloroplastic ACCase activity, and this was completely reversed in the presence of 2-oxoglutarate, pyruvate, or oxaloacetate. The inhibitory effect was PII-dose dependent and appeared to be PII-specific because ACCase activity was not altered in the presence of other tested proteins. PII decreased the V(max) of the ACCase reaction without altering the K(m) for acetyl-CoA. These data show that PII function has evolved between bacterial and plant systems to control the carbon metabolism pathway of fatty acid synthesis in plastids. PMID- 20018656 TI - Identification of MIR390a precursor processing-defective mutants in Arabidopsis by direct genome sequencing. AB - Transacting siRNA (tasiRNA) biogenesis in Arabidopsis is initiated by microRNA (miRNA) -guided cleavage of primary transcripts. In the case of TAS3 tasiRNA formation, ARGONAUTE7 (AGO7)-miR390 complexes interact with primary transcripts at two sites, resulting in recruitment of RNA-DEPENDENT RNA POLYMERASE6 for dsRNA biosynthesis. An extensive screen for Arabidopsis mutants with specific defects in TAS3 tasiRNA biogenesis or function was done. This yielded numerous ago7 mutants, one dcl4 mutant, and two mutants that accumulated low levels of miR390. A direct genome sequencing-based approach to both map and rapidly identify one of the latter mutant alleles was developed. This revealed a G-to-A point mutation (mir390a-1) that was calculated to stabilize a relatively nonpaired region near the base of the MIR390a foldback, resulting in misprocessing of the miR390/miR390* duplex and subsequent reduced TAS3 tasiRNA levels. Directed substitutions, as well as analysis of variation at paralogous miR390-generating loci (MIR390a and MIR390b), indicated that base pair properties and nucleotide identity within a region 4-6 bases below the miR390/miR390* duplex region contributed to the efficiency and accuracy of precursor processing. PMID- 20018657 TI - A role for suppressed incisor cuspal morphogenesis in the evolution of mammalian heterodont dentition. AB - Changes in tooth shape have played a major role in vertebrate evolution with modification of dentition allowing an organism to adapt to new feeding strategies. The current view is that molar teeth evolved from simple conical teeth, similar to canines, by progressive addition of extra "cones" to form progressively complex multicuspid crowns. Mammalian incisors, however, are neither conical nor multicuspid, and their evolution is unclear. We show that hypomorphic mutation of a cell surface receptor, Lrp4, which modulates multiple signaling pathways, produces incisors with grooved enamel surfaces that exhibit the same molecular characteristics as the tips of molar cusps. Mice with a null mutation of Lrp4 develop extra cusps on molars and have incisors that exhibit clear molar-like cusp and root morphologies. Molecular analysis identifies misregulation of Shh and Bmp signaling in the mutant incisors and suggests an uncoupling of the processes of tooth shape determination and morphogenesis. Incisors thus possess a developmentally suppressed, cuspid crown-like morphogenesis program similar to that in molars that is revealed by loss of Lrp4 activity. Several mammalian species naturally possess multicuspid incisors, suggesting that mammals have the capacity to form multicuspid teeth regardless of location in the oral jaw. Localized loss of enamel may thus have been an intermediary step in the evolution of cusps, both of which use Lrp4-mediated signaling. PMID- 20018658 TI - Building biological memory by linking positive feedback loops. AB - A common topology found in many bistable genetic systems is two interacting positive feedback loops. Here we explore how this relatively simple topology can allow bistability over a large range of cellular conditions. On the basis of theoretical arguments, we predict that nonlinear interactions between two positive feedback loops can produce an ultrasensitive response that increases the range of cellular conditions at which bistability is observed. This prediction was experimentally tested by constructing a synthetic genetic circuit in Escherichia coli containing two well-characterized positive feedback loops, linked in a coherent fashion. The concerted action of both positive feedback loops resulted in bistable behavior over a broad range of inducer concentrations; when either of the feedback loops was removed, the range of inducer concentrations at which the system exhibited bistability was decreased by an order of magnitude. Furthermore, bistability of the system could be tuned by altering growth conditions that regulate the contribution of one of the feedback loops. Our theoretical and experimental work shows how linked positive feedback loops may produce the robust bistable responses required in cellular networks that regulate development, the cell cycle, and many other cellular responses. PMID- 20018659 TI - A genomewide study identifies the Wnt signaling pathway as a major target of p53 in murine embryonic stem cells. AB - Both p53 and the Wnt signaling pathway play important roles in regulating the differentiation of mouse embryonic stem cells (mESCs). However, it is not known whether they directly and/or functionally crosstalk in mESCs. Here we report a surprising antidifferentiation function of p53 in mESCs through directly regulating the Wnt signaling pathway. A chromatin-immunoprecipitation-based microarray (ChIP-chip) and gene expression microarray assays reveal that the Wnt signaling pathway is significantly (P value, 0.000048) overrepresented in p53 regulated genes in mESCs. The expression of five Wnt ligand genes is robustly induced by various genotoxic and nongenotoxic insults in a p53-dependent manner. Moreover, the induction of these Wnt genes is greatly attenuated in mouse embryonic fibroblast (MEF) cells and ESC-derived neural stem/progenitor cells, suggesting that the induction is mESC specific. It is established that the activation of the Wnt signaling pathway inhibits the differentiation of mESCs. Consistent with this notion, we detected an antidifferentiation activity from the conditioned medium (CM) collected from UV (UV)-treated mESCs. This antidifferentiation activity can be lowered by either the addition of Wnt antagonists into the CM or the reduction of p53 levels in UV-treated mESCs. Therefore, reminiscent of its dual functions on death and survival in somatic cells, p53 appears to regulate both prodifferentiation and antidifferentiation programs in mESCs. Our findings uncover a direct and functional connection between p53 and the Wnt signaling pathway, and expand the catalog of p53 regulated genes in mESCs. PMID- 20018660 TI - Homeostatic imbalance of regulatory and effector T cells due to IL-2 deprivation amplifies murine lupus. AB - The origins and consequences of a regulatory T cell (Treg) disorder in systemic lupus erythematosus (SLE) are poorly understood. In the (NZBxNZW) F(1) mouse model of lupus, we found that CD4(+)Foxp3(+) Treg failed to maintain a competitive pool size in the peripheral lymphoid organs resulting in a progressive homeostatic imbalance of CD4(+)Foxp3(+) Treg and CD4(+)Foxp3(-) conventional T cells (Tcon). In addition, Treg acquired phenotypic changes that are reminiscent of IL-2 deficiency concomitantly to a progressive decline in IL-2 producing Tcon and an increase in activated, IFN-gamma-producing effector Tcon. Nonetheless, Treg from lupus-prone mice were functionally intact and capable to influence the course of disease. Systemic reduction of IL-2 levels early in disease promoted Tcon hyperactivity, induced the imbalance of Treg and effector Tcon, and strongly accelerated disease progression. In contrast, administration of IL-2 partially restored the balance of Treg and effector Tcon by promoting the homeostatic proliferation of endogenous Treg and impeded the progression of established disease. Thus, an acquired and self-amplifying disruption of the Treg IL-2 axis contributed essentially to Tcon hyperactivity and the development of murine lupus. The reversibility of this homeostatic Treg disorder provides promising approaches for the treatment of SLE. PMID- 20018661 TI - Identification of a small-molecule inhibitor of the PICK1 PDZ domain that inhibits hippocampal LTP and LTD. AB - Proteins containing PSD-95/Discs-large/ZO-1 homology (PDZ) domains play key roles in the assembly and regulation of cellular signaling pathways and represent putative targets for new pharmacotherapeutics. Here we describe the first small molecule inhibitor (FSC231) of the PDZ domain in protein interacting with C kinase 1 (PICK1) identified by a screening of approximately 44,000 compounds in a fluorescent polarization assay. The inhibitor bound the PICK1 PDZ domain with an affinity similar to that observed for endogenous peptide ligands (K(i) approximately 10.1 microM). Mutational analysis, together with computational docking of the compound in simulations starting from the PDZ domain structure, identified the binding mode of FSC231. The specificity of FSC231 for the PICK1 PDZ domain was supported by the lack of binding to PDZ domains of postsynaptic density protein 95 (PSD-95) and glutamate receptor interacting protein 1 (GRIP1). Pretreatment of cultured hippocampal neurons with FSC231 inhibited coimmunopreciptation of the AMPA receptor GluR2 subunit with PICK1. In agreement with inhibiting the role of PICK1 in GluR2 trafficking, FSC231 accelerated recycling of pHluorin-tagged GluR2 in hippocampal neurons after internalization in response to NMDA receptor activation. FSC231 blocked the expression of both long-term depression and long-term potentiation in hippocampal CA1 neurons from acute slices, consistent with inhibition of the bidirectional function of PICK1 in synaptic plasticity. Given the proposed role of the PICK1/AMPA receptor interaction in neuropathic pain, excitotoxicity, and cocaine addiction, FSC231 might serve as a lead in the future development of new therapeutics against these conditions. PMID- 20018662 TI - Delayed wave of c-Fos expression in the dorsal hippocampus involved specifically in persistence of long-term memory storage. AB - Memory formation is a temporally graded process during which transcription and translation steps are required in the first hours after acquisition. Although persistence is a key characteristic of memory storage, its mechanisms are scarcely characterized. Here, we show that long-lasting but not short-lived inhibitory avoidance long-term memory is associated with a delayed expression of c-Fos in the hippocampus. Importantly, this late wave of c-Fos is necessary for maintenance of inhibitory avoidance long-term storage. Moreover, inhibition of transcription in the dorsal hippocampus 24 h after training hinders persistence but not formation of long-term storage. These findings indicate that a delayed phase of transcription is essential for maintenance of a hippocampus-dependent memory trace. Our results support the hypothesis that recurrent rounds of consolidation-like events take place late after learning in the dorsal hippocampus to maintain memories. PMID- 20018663 TI - Cleistogamous flowering in barley arises from the suppression of microRNA-guided HvAP2 mRNA cleavage. AB - The cleistogamous flower sheds its pollen before opening, forcing plants with this habit to be almost entirely autogamous. Cleistogamy also provides a means of escape from cereal head blight infection and minimizes pollen-mediated gene flow. The lodicule in cleistogamous barley is atrophied. We have isolated cleistogamy 1 (Cly1) by positional cloning and show that it encodes a transcription factor containing two AP2 domains and a putative microRNA miR172 targeting site, which is an ortholog of Arabidopsis thaliana AP2. The expression of Cly1 was concentrated within the lodicule primordia. We established a perfect association between a synonymous nucleotide substitution at the miR172 targeting site and cleistogamy. Cleavage of mRNA directed by miR172 was detectable only in a noncleistogamous background. We conclude that the miR172-derived down-regulation of Cly1 promotes the development of the lodicules, thereby ensuring noncleistogamy, although the single nucleotide change at the miR172 targeting site results in the failure of the lodicules to develop properly, producing the cleistogamous phenotype. PMID- 20018664 TI - Somatic tetraploidy in specific chick retinal ganglion cells induced by nerve growth factor. AB - A subset of neurons in the normal vertebrate nervous system contains double the normal amount of DNA in their nuclei. These neurons are all thought to derive from aberrant mitoses in neuronal precursor cells. Here we show that endogenous NGF induces DNA replication in a subpopulation of differentiating chick retinal ganglion cells that express both the neurotrophin receptor p75 and the E2F1 transcription factor, but that lack the retinoblastoma protein. Many of these neurons avoid G2/M transition and remain alive in the retina as tetraploid cells with large cell somas and extensive dendritic trees, and most of them express beta2 nicotinic acetylcholine receptor subunits, a specific marker of retinal ganglion cells innervating lamina F in the stratum-griseum-et-fibrosum superficiale of the tectal cortex. Tetraploid neurons were also observed in the adult mouse retina. Thus, a developmental program leading to somatic tetraploidy in specific retinal neurons exists in vertebrates. This program might occur in other vertebrate neurons during normal or pathological situations. PMID- 20018665 TI - Ebolavirus VP35 uses a bimodal strategy to bind dsRNA for innate immune suppression. AB - Ebolavirus causes a severe hemorrhagic fever and is divided into five distinct species, of which Reston ebolavirus is uniquely nonpathogenic to humans. Disease caused by ebolavirus is marked by early immunosuppression of innate immune signaling events, involving silencing and sequestration of double-stranded RNA (dsRNA) by the viral protein VP35. Here we present unbound and dsRNA-bound crystal structures of the dsRNA-binding domain of Reston ebolavirus VP35. The structures show that VP35 forms an unusual, asymmetric dimer on dsRNA binding, with each of the monomers binding dsRNA in a different way: one binds the backbone whereas the other caps the terminus. Additional SAXS, DXMS, and dsRNA binding experiments presented here support a model of cooperative dsRNA recognition in which binding of the first monomer assists binding of the next monomer of the oligomeric VP35 protein. This work illustrates how ebolavirus VP35 could mask key recognition sites of molecules such as RIG-I, MDA-5, and Dicer to silence viral dsRNA in infection. PMID- 20018666 TI - Laser microdissection of Arabidopsis cells at the powdery mildew infection site reveals site-specific processes and regulators. AB - To elucidate host processes and components required for the sustained growth and reproduction of the obligate biotrophic fungus Golovinomyces orontii on Arabidopsis thaliana, laser microdissection was used to isolate cells at the site of infection at 5 days postinfection for downstream global Arabidopsis expression profiling. Site-specific profiling increased sensitivity dramatically, allowing us to identify specific host processes, process components, and their putative regulators hidden in previous whole-leaf global expression analyses. For example, 67 transcription factors exhibited altered expression at the powdery mildew (PM) infection site, with subsets of these playing known or inferred roles in photosynthesis, cold/dehydration responses, defense, auxin signaling, and the cell cycle. Using integrated informatics analyses, we constructed putative regulatory networks for a subset of these processes and provided strong support for host cell cycle modulation at the PM infection site. Further experimentation revealed induced host endoreduplication occurred exclusively at the infection site and led us to identify MYB3R4 as a transcriptional regulator of this process. Induced endoreduplication was abrogated in myb3r4 mutants, and G. orontii growth and reproduction were reduced. This suggests that, by increasing gene copy number, localized endoreduplication serves as a mechanism to meet the enhanced metabolic demands imposed by the fungus, which acquires all its nutrients from the plant host. PMID- 20018667 TI - Bicarbonate-sensing soluble adenylyl cyclase is an essential sensor for acid/base homeostasis. AB - pH homeostasis is essential for life, yet it remains unclear how animals sense their systemic acid/base (A/B) status. Soluble adenylyl cyclase (sAC) is an evolutionary conserved signaling enzyme that produces the second messenger cAMP in response to bicarbonate ions (HCO(3)(-)). We cloned the sAC ortholog from the dogfish, a shark that regulates blood A/B by absorbing and secreting protons (H(+)) and HCO(3)(-) at its gills. Similar to mammalian sAC, dogfish soluble adenylyl cyclase (dfsAC) is activated by HCO(3)(-) and can be inhibited by two structurally and mechanistically distinct small molecule inhibitors. dfsAC is expressed in the gill epithelium, where the subset of base-secreting cells resides. Injection of inhibitors into animals under alkaline stress confirmed that dfsAC is essential for maintaining systemic pH and HCO(3)(-) levels in the whole organism. One of the downstream effects of dfsAC is to promote the insertion of vacuolar proton pumps into the basolateral membrane to absorb H(+) into the blood. sAC orthologs are present throughout metazoans, and mammalian sAC is expressed in A/B regulatory organs, suggesting that systemic A/B sensing via sAC is widespread in the animal kingdom. PMID- 20018668 TI - Recombination-induced tag exchange to track old and new proteins. AB - The dynamic behavior of proteins is critical for cellular homeostasis. However, analyzing dynamics of proteins and protein complexes in vivo has been difficult. Here we describe recombination-induced tag exchange (RITE), a genetic method that induces a permanent epitope-tag switch in the coding sequence after a hormone induced activation of Cre recombinase. The time-controlled tag switch provides a unique ability to detect and separate old and new proteins in time and space, which opens up opportunities to investigate the dynamic behavior of proteins. We validated the technology by determining exchange of endogenous histones in chromatin by biochemical methods and by visualizing and quantifying replacement of old by new proteasomes in single cells by microscopy. RITE is widely applicable and allows probing spatiotemporal changes in protein properties by multiple methods. PMID- 20018669 TI - Whole-proteome phylogeny of prokaryotes by feature frequency profiles: An alignment-free method with optimal feature resolution. AB - We present a whole-proteome phylogeny of prokaryotes constructed by comparing feature frequency profiles (FFPs) of whole proteomes. Features are l-mers of amino acids, and each organism is represented by a profile of frequencies of all features. The selection of feature length is critical in the FFP method, and we have developed a procedure for identifying the optimal feature lengths for inferring the phylogeny of prokaryotes, strictly speaking, a proteome phylogeny. Our FFP trees are constructed with whole proteomes of 884 prokaryotes, 16 unicellular eukaryotes, and 2 random sequences. To highlight the branching order of major groups, we present a simplified proteome FFP tree of monophyletic class or phylum with branch support. In our whole-proteome FFP trees (i) Archaea, Bacteria, Eukaryota, and a random sequence outgroup are clearly separated; (ii) Archaea and Bacteria form a sister group when rooted with random sequences; (iii) Planctomycetes, which possesses an intracellular membrane compartment, is placed at the basal position of the Bacteria domain; (iv) almost all groups are monophyletic in prokaryotes at most taxonomic levels, but many differences in the branching order of major groups are observed between our proteome FFP tree and trees built with other methods; and (v) previously "unclassified" genomes may be assigned to the most likely taxa. We describe notable similarities and differences between our FFP trees and those based on other methods in grouping and phylogeny of prokaryotes. PMID- 20018670 TI - A muscle-specific transgenic reporter line of the sea anemone, Nematostella vectensis. AB - The sea anemone, Nematostella vectensis, has become an attractive new model organism for comparative genomics and evolutionary developmental biology. Over the last few years, many genes have been isolated and their expression patterns studied to gain insight into their function. More recently, functional tools have been developed to manipulate gene function; however, most of these approaches rely on microinjection and are limited to early stages of development. Transgenic lines would significantly enhance the tractability of the system. In particular, the study of gene- or tissue-specific promoters would be most useful. Here we report the stable establishment of a transgenic line using the I-SceI meganuclease system to facilitate integration into the genome. We isolated a 1.6 kb fragment of the regulatory upstream region of the Myosin Heavy Chain1 (MyHC1) gene and found that the transgene is specifically expressed in the retractor and tentacle muscles of Nematostella polyps, faithfully reproducing the expression of the endogenous MyHC1 gene. This demonstrates that the 1.6-kb fragment contains all of the regulatory elements necessary to drive correct expression and suggests that retractor and tentacle muscles in Nematostella are distinct from other myoepithelial cells. The transgene is transmitted through the germline at high frequency, and G(1) transgenic polyps have only one integration site. The relatively high frequency of transgenesis, in combination with gene- or tissue specific promoters, will foster experimental possibilities for studying in vivo gene functions in gene regulatory networks and developmental processes in the nonbilaterian sea anemone, Nematostella vectensis. PMID- 20018671 TI - Diversity and evolution of 11 innate immune genes in Bos taurus taurus and Bos taurus indicus cattle. AB - The Toll-like receptor (TLR) and peptidoglycan recognition protein 1 (PGLYRP1) genes play key roles in the innate immune systems of mammals. While the TLRs recognize a variety of invading pathogens and induce innate immune responses, PGLYRP1 is directly microbicidal. We used custom allele-specific assays to genotype and validate 220 diallelic variants, including 54 nonsynonymous SNPs in 11 bovine innate immune genes (TLR1-TLR10, PGLYRP1) for 37 cattle breeds. Bayesian haplotype reconstructions and median joining networks revealed haplotype sharing between Bos taurus taurus and Bos taurus indicus breeds at every locus, and we were unable to differentiate between the specialized B. t. taurus beef and dairy breeds, despite an average polymorphism density of one locus per 219 bp. Ninety-nine tagSNPs and one tag insertion-deletion polymorphism were sufficient to predict 100% of the variation at all 11 innate immune loci in both subspecies and their hybrids, whereas 58 tagSNPs captured 100% of the variation at 172 loci in B. t. taurus. PolyPhen and SIFT analyses of nonsynonymous SNPs encoding amino acid replacements indicated that the majority of these substitutions were benign, but up to 31% were expected to potentially impact protein function. Several diversity-based tests provided support for strong purifying selection acting on TLR10 in B. t. taurus cattle. These results will broadly impact efforts related to bovine translational genomics. PMID- 20018672 TI - VGLUT2 in dopamine neurons is required for psychostimulant-induced behavioral activation. AB - The "One neuron-one neurotransmitter" concept has been challenged frequently during the last three decades, and the coexistence of neurotransmitters in individual neurons is now regarded as a common phenomenon. The functional significance of neurotransmitter coexistence is, however, less well understood. Several studies have shown that a subpopulation of dopamine (DA) neurons in the ventral tegmental area (VTA) expresses the vesicular glutamate transporter 2 (VGLUT2) and has been suggested to use glutamate as a cotransmitter. The VTA dopamine neurons project to limbic structures including the nucleus accumbens, and are involved in mediating the motivational and locomotor activating effects of psychostimulants. To determine the functional role of glutamate cotransmission by these neurons, we deleted VGLUT2 in DA neurons by using a conditional gene targeting approach in mice. A DAT-Cre/Vglut2Lox mouse line (Vglut2(f/f;DAT-Cre) mice) was produced and analyzed by in vivo amperometry as well as by several behavioral paradigms. Although basal motor function was normal in the Vglut2(f/f;DAT-Cre) mice, their risk-taking behavior was altered. Interestingly, in both home-cage and novel environments, the gene targeted mice showed a greatly blunted locomotor response to the psychostimulant amphetamine, which acts via the midbrain DA system. Our results show that VGLUT2 expression in DA neurons is required for normal emotional reactivity as well as for psychostimulant-mediated behavioral activation. PMID- 20018673 TI - miRNA-processing enzyme Dicer is necessary for cardiac outflow tract alignment and chamber septation. AB - MicroRNAs (miRNAs) have previously been implicated in a number of developmental processes, including development of the ventricular myocardium of the heart. To determine what, if any, additional roles miRNAs play in cardiogenesis, we deleted the miRNA-processing enzyme Dicer specifically in the developing murine heart. Embryos lacking cardiac Dicer lived longer than reported in previous studies using different alleles to remove cardiac Dicer activity and displayed a highly penetrant phenotype of double outlet right ventricle with a concurrent ventricular septal defect. Before the defect's onset, Pitx2c and Sema3c, both required for outflow tract morphogenesis, were up-regulated in Dicer-deficient hearts. Interestingly, mesenchymal apoptosis in the outflow tract normally required for outflow tract alignment was greatly decreased in the mutants, likely contributing directly to the observed phenotype. In sum, we demonstrate here a specific developmental process, that of outflow tract morphogenesis, being hindered by the deletion of miRNAs during cardiogenesis. PMID- 20018674 TI - Involvement of the Toll-like receptor 4 pathway and use of TNF-alpha antagonists for treatment of the mucopolysaccharidoses. AB - Enzyme replacement therapy is currently available for three of the mucopolysaccharidoses (MPSs) but has limited effects on the skeletal lesions. We investigated the involvement of the Toll-like receptor 4 (TLR4) signaling pathway in the pathogenesis of MPS bone and joint disease, and the use of the anti-TNF alpha drug, Remicade (Centocor, Inc.), for treatment. TLR4 KO (TLR4(lps-/-)) mice were interbred with MPS VII mice to produce double-KO (DKO) animals. The DKO mice had longer and thinner faces and longer femora as revealed by micro-computed tomography analysis compared with MPS VII mice. Histological analyses also revealed more organized and thinner growth plates. The serum levels of TNF-alpha were normalized in the DKO animals, and the levels of phosphorylated STAT1 and STAT3 in articular chondrocytes were corrected. These findings led us to evaluate the effects of Remicade in MPS VI rats. When initiated at 1 month of age, i.v. treatment prevented the elevation of TNF-alpha, receptor activator of NF-kappaB, and other inflammatory molecules not only in the blood but in articular chondrocytes and fibroblast-like synoviocytes (FLSs). Treatment of 6-month-old animals also reduced the levels of these molecules to normal. The number of apoptotic articular chondrocytes in MPS VI rats was similarly reduced, with less infiltration of synovial tissue into the underlying bone. These studies revealed the important role of TLR4 signaling in MPS bone and joint disease and suggest that targeting TNF-alpha may have positive therapeutic effects. PMID- 20018675 TI - Electrokinesis is a microbial behavior that requires extracellular electron transport. AB - We report a previously undescribed bacterial behavior termed electrokinesis. This behavior was initially observed as a dramatic increase in cell swimming speed during reduction of solid MnO(2) particles by the dissimilatory metal-reducing bacterium Shewanella oneidensis MR-1. The same behavioral response was observed when cells were exposed to small positive applied potentials at the working electrode of a microelectrochemical cell and could be tuned by adjusting the potential on the working electrode. Electrokinesis was found to be different from both chemotaxis and galvanotaxis but was absent in mutants defective in electron transport to solid metal oxides. Using in situ video microscopy and cell tracking algorithms, we have quantified the response for different strains of Shewanella and shown that the response correlates with current-generating capacity in microbial fuel cells. The electrokinetic response was only exhibited by a subpopulation of cells closest to the MnO(2) particles or electrodes. In contrast, the addition of 1 mM 9,10-anthraquinone-2,6-disulfonic acid, a soluble electron shuttle, led to increases in motility in the entire population. Electrokinesis is defined as a behavioral response that requires functional extracellular electron transport and that is observed as an increase in cell swimming speeds and lengthened paths of motion that occur in the proximity of a redox active mineral surface or the working electrode of an electrochemical cell. PMID- 20018676 TI - On implementing maximum economic yield in commercial fisheries. AB - Economists have long argued that a fishery that maximizes its economic potential usually will also satisfy its conservation objectives. Recently, maximum economic yield (MEY) has been identified as a primary management objective for Australian fisheries and is under consideration elsewhere. However, first attempts at estimating MEY as an actual management target for a real fishery (rather than a conceptual or theoretical exercise) have highlighted some substantial complexities generally unconsidered by fisheries economists. Here, we highlight some of the main issues encountered in our experience and their implications for estimating and transitioning to MEY. Using a bioeconomic model of an Australian fishery for which MEY is the management target, we note that unconstrained optimization may result in effort trajectories that would not be acceptable to industry or managers. Different assumptions regarding appropriate constraints result in different outcomes, each of which may be considered a valid MEY. Similarly, alternative treatments of prices and costs may result in differing estimates of MEY and their associated effort trajectories. To develop an implementable management strategy in an adaptive management framework, a set of assumptions must be agreed among scientists, economists, and industry and managers, indicating that operationalizing MEY is not simply a matter of estimating the numbers but requires strong industry commitment and involvement. PMID- 20018677 TI - Long-term nutrient enrichment decouples predator and prey production. AB - Increased nutrient mobilization by human activities represents one of the greatest threats to global ecosystems, but its effects on ecosystem productivity can differ depending on food web structure. When this structure facilitates efficient energy transfers to higher trophic levels, evidence from previous large scale enrichments suggests that nutrients can stimulate the production of multiple trophic levels. Here we report results from a 5-year continuous nutrient enrichment of a forested stream that increased primary consumer production, but not predator production. Because of strong positive correlations between predator and prey production (evidence of highly efficient trophic transfers) under reference conditions, we originally predicted that nutrient enrichment would stimulate energy flow to higher trophic levels. However, enrichment decoupled this strong positive correlation and produced a nonlinear relationship between predator and prey production. By increasing the dominance of large-bodied predator-resistant prey, nutrient enrichment truncated energy flow to predators and reduced food web efficiency. This unexpected decline in food web efficiency indicates that nutrient enrichment, a ubiquitous threat to aquatic ecosystems, may have unforeseen and unpredictable effects on ecosystem structure and productivity. PMID- 20018678 TI - Plant flotillins are required for infection by nitrogen-fixing bacteria. AB - To establish compatible rhizobial-legume symbioses, plant roots support bacterial infection via host-derived infection threads (ITs). Here, we report the requirement of plant flotillin-like genes (FLOTs) in Sinorhizobium meliloti infection of its host legume Medicago truncatula. Flotillins in other organisms have roles in viral pathogenesis, endocytosis, and membrane shaping. We identified seven FLOT genes in the M. truncatula genome and show that two, FLOT2 and FLOT4, are strongly up-regulated during early symbiotic events. This up regulation depends on bacterial Nod Factor and the plant's ability to perceive Nod Factor. Microscopy data suggest that M. truncatula FLOT2 and FLOT4 localize to membrane microdomains. Upon rhizobial inoculation, FLOT4 uniquely becomes localized to the tips of elongating root hairs. Silencing FLOT2 and FLOT4 gene expression reveals a nonredundant requirement for both genes in IT initiation and nodule formation. FLOT4 is uniquely required for IT elongation, and FLOT4 localizes to IT membranes. This work reveals a critical role for plant flotillins in symbiotic bacterial infection. PMID- 20018679 TI - Sinorhizobium meliloti phospholipase C required for lipid remodeling during phosphorus limitation. AB - Rhizobia are Gram-negative soil bacteria able to establish nitrogen-fixing root nodules with their respective legume host plants. Besides phosphatidylglycerol, cardiolipin, and phosphatidylethanolamine, rhizobial membranes contain phosphatidylcholine (PC) as a major membrane lipid. Under phosphate-limiting conditions of growth, some bacteria replace their membrane phospholipids with lipids lacking phosphorus. In Sinorhizobium meliloti, these phosphorus-free lipids are sulfoquinovosyl diacylglycerol, ornithine-containing lipid, and diacylglyceryl trimethylhomoserine (DGTS). Pulse-chase experiments suggest that the zwitterionic phospholipids phosphatidylethanolamine and PC act as biosynthetic precursors of DGTS under phosphorus-limiting conditions. A S. meliloti mutant, deficient in the predicted phosphatase SMc00171 was unable to degrade PC or to form DGTS in a similar way as the wild type. Cell-free extracts of Escherichia coli, in which SMc00171 had been expressed, convert PC to phosphocholine and diacylglycerol, showing that SMc00171 functions as a phospholipase C. Diacylglycerol , in turn, is the lipid anchor from which biosynthesis is initiated during the formation of the phosphorus-free membrane lipid DGTS. Inorganic phosphate can be liberated from phosphocholine. These data suggest that, in S. meliloti under phosphate-limiting conditions, membrane phospholipids provide a pool for metabolizable inorganic phosphate, which can be used for the synthesis of other essential phosphorus-containing biomolecules. This is an example of an intracellular phospholipase C in a bacterial system; however, the ability to degrade endogenous preexisting membrane phospholipids as a source of phosphorus may be a general property of Gram-negative soil bacteria. PMID- 20018680 TI - A FOXA1-binding enhancer regulates Hoxb13 expression in the prostate gland. AB - Hoxb13 is robustly transcribed in derivatives of posterior endoderm including the colon, rectum, and the prostate gland. Transcriptional activity in the prostate persists unabated under conditions of androgen deprivation and throughout the course of disease progression in a mouse prostate cancer model. To elucidate the molecular basis of prostate-restricted transcriptional activation of Hoxb13, a bacterial artificial chromosome (BAC)-based reporter gene deletion analysis was performed in transgenic mice. Two regions downstream of the Hoxb13 coding region were found to be required to support transcriptional activity in the prostate but were completely dispensable for expression in the colon and rectum. Bioinformatic analyses of one region identified a 37-bp element conserved in mammals. This element, which bears two potential binding sites for Forkhead class transcription factors, is occupied by FOXA1 in a human prostate cancer cell line. Precise replacement of this enhancer with an extended LoxP site in the context of a 218,555-bp BAC reporter nearly extinguished Hoxb13-mediated transcriptional activity in the mouse prostate. These data demonstrate that FOXA1 directly regulates HOXB13 in human prostate epithelial cells, and show that this prostate specific regulatory mechanism is conserved in mice. PMID- 20018681 TI - Evolution in health and medicine Sackler colloquium: a public choice framework for controlling transmissible and evolving diseases. AB - Control measures used to limit the spread of infectious disease often generate externalities. Vaccination for transmissible diseases can reduce the incidence of disease even among the unvaccinated, whereas antimicrobial chemotherapy can lead to the evolution of antimicrobial resistance and thereby limit its own effectiveness over time. We integrate the economic theory of public choice with mathematical models of infectious disease to provide a quantitative framework for making allocation decisions in the presence of these externalities. To illustrate, we present a series of examples: vaccination for tetanus, vaccination for measles, antibiotic treatment of otitis media, and antiviral treatment of pandemic influenza. PMID- 20018682 TI - Homozygous frameshift mutation in TMCO1 causes a syndrome with craniofacial dysmorphism, skeletal anomalies, and mental retardation. AB - We identified an autosomal recessive condition in 11 individuals in the Old Order Amish of northeastern Ohio. The syndrome was characterized by distinctive craniofacial dysmorphism, skeletal anomalies, and mental retardation. The typical craniofacial dysmorphism included brachycephaly, highly arched bushy eyebrows, synophrys, long eyelashes, low-set ears, microdontism of primary teeth, and generalized gingival hyperplasia, whereas Sprengel deformity of scapula, fusion of spine, rib abnormities, pectus excavatum, and pes planus represented skeletal anomalies. The genome-wide homozygosity mapping using six affected individuals localized the disease gene to a 3.3-Mb region on chromosome 1q23.3-q24.1. Candidate gene sequencing identified a homozygous frameshift mutation, c.139_140delAG, in the transmembrane and coiled-coil domains 1 (TMCO1) gene, as the pathogenic change in all affected members of the extended pedigree. This mutation is predicted to result in a severely truncated protein (p.Ser47Ter) of only one-fourth the original length. The TMCO1 gene product is a member of DUF841 superfamily of several eukaryotic proteins with unknown function. The gene has highly conserved amino acid sequence and is universally expressed in all human tissues examined. The high degree of conservation and the ubiquitous expression pattern in human adult and fetal tissues suggest a critical role for TMCO1. This report shows a TMCO1 sequence variant being associated with a genetic disorder in human. We propose "TMCO1 defect syndrome" as the name of this condition. PMID- 20018683 TI - Recognition memory signals in the macaque hippocampus. AB - The hippocampus plays a critical role in recognition memory in both monkeys and humans. However, neurophysiological studies have rarely reported recognition memory signals among hippocampal neurons. The majority of these previous studies used variants of the delayed match-to-sample task; however, studies of the effects of hippocampal damage in monkey and humans have shown that another task of recognition memory, the visual paired-comparison, or visual preferential looking task (VPLT), is more sensitive to hippocampal damage than the delayed matching tasks. Accordingly, to examine possible recognition memory signals in the hippocampus, we recorded the activity of 131 hippocampal neurons in two monkeys performing the VPLT. Eighty-eight neurons (67%) responded significantly to stimulus presentation relative to the baseline prestimulus period. A substantial proportion of these visually responsive neurons (36%) showed significant firing-rate modulations that reflected whether stimuli were novel or familiar. Additionally, these firing-rate modulations were correlated with recognition memory performance on the VPLT such that larger modulations by stimulus novelty were associated with better performance. Together, these results provide evidence for a neural signal in the hippocampus that may support recognition memory performance. PMID- 20018684 TI - Direct rod input to cone BCs and direct cone input to rod BCs challenge the traditional view of mammalian BC circuitry. AB - Bipolar cells are the central neurons of the retina that transmit visual signals from rod and cone photoreceptors to third-order neurons in the inner retina and the brain. A dogma set forth by early anatomical studies is that bipolar cells in mammalian retinas receive segregated rod/cone synaptic inputs (either from rods or from cones), and here, we present evidence that challenges this traditional view. By analyzing light-evoked cation currents from morphologically identified depolarizing bipolar cells (DBCs) in the wild-type and three pathway-specific knockout mice (rod transducin knockout [Tralpha(-/-)], connexin36 knockout [Cx36( /-)], and transcription factor beta4 knockout [Bhlhb4(-/-)]), we show that a subpopulation of rod DBCs (DBC(R2)s) receives substantial input directly from cones and a subpopulation of cone DBCs (DBC(C1)s) receives substantial input directly from rods. These results provide evidence of the existence of functional rod-DBC(C) and cone-DBC(R) synaptic pathways in the mouse retina as well as the previously proposed rod hyperpolarizing bipolar-cells pathway. This is grounds for revising the mammalian rod/cone bipolar cell dogma. PMID- 20018685 TI - Gene amplification confers glyphosate resistance in Amaranthus palmeri. AB - The herbicide glyphosate became widely used in the United States and other parts of the world after the commercialization of glyphosate-resistant crops. These crops have constitutive overexpression of a glyphosate-insensitive form of the herbicide target site gene, 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS). Increased use of glyphosate over multiple years imposes selective genetic pressure on weed populations. We investigated recently discovered glyphosate resistant Amaranthus palmeri populations from Georgia, in comparison with normally sensitive populations. EPSPS enzyme activity from resistant and susceptible plants was equally inhibited by glyphosate, which led us to use quantitative PCR to measure relative copy numbers of the EPSPS gene. Genomes of resistant plants contained from 5-fold to more than 160-fold more copies of the EPSPS gene than did genomes of susceptible plants. Quantitative RT-PCR on cDNA revealed that EPSPS expression was positively correlated with genomic EPSPS relative copy number. Immunoblot analyses showed that increased EPSPS protein level also correlated with EPSPS genomic copy number. EPSPS gene amplification was heritable, correlated with resistance in pseudo-F(2) populations, and is proposed to be the molecular basis of glyphosate resistance. FISH revealed that EPSPS genes were present on every chromosome and, therefore, gene amplification was likely not caused by unequal chromosome crossing over. This occurrence of gene amplification as an herbicide resistance mechanism in a naturally occurring weed population is particularly significant because it could threaten the sustainable use of glyphosate-resistant crop technology. PMID- 20018686 TI - A receptor-like cytoplasmic kinase, BIK1, associates with a flagellin receptor complex to initiate plant innate immunity. AB - Plants and animals rely on innate immunity to prevent infections by detection of microbe-associated molecular patterns (MAMPs) through pattern-recognition receptors (PRRs). The plant PRR FLS2, a leucine-rich repeat-receptor kinase, recognizes bacterial flagellin and initiates immune signaling by association with another leucine-rich repeat-receptor-like kinase, BAK1. It remains unknown how the FLS2/BAK1 receptor complex activates intracellular signaling cascades. Here we identified the receptor-like cytoplasmic kinase BIK1 that is rapidly phosphorylated upon flagellin perception, depending on both FLS2 and BAK1. BIK1 associates with FLS2 and BAK1 in vivo and in vitro. BIK1 is phosphorylated by BAK1, and BIK1 also directly phosphorylates BAK1 and FLS2 in vitro. The flagellin phosphorylation site Thr(237) of BIK1 is required for its phosphorylation on BAK1 and FLS2, suggesting that BIK1 is likely first phosphorylated upon flagellin perception and subsequently transphosphorylates FLS2/BAK1 to propagate flagellin signaling. Importantly, bik1 mutants are compromised in diverse flagellin mediated responses and immunity to the nonpathogenic bacterial infection. Thus, BIK1 is an essential component in MAMP signal transduction, which links the MAMP receptor complex to downstream intracellular signaling. PMID- 20018687 TI - Defined factors induce reprogramming of gastrointestinal cancer cells. AB - Although cancer is a disease with genetic and epigenetic origins, the possible effects of reprogramming by defined factors remain to be fully understood. We studied the effects of the induction or inhibition of cancer-related genes and immature status-related genes whose alterations have been reported in gastrointestinal cancer cells. Retroviral-mediated introduction of induced pluripotent stem (iPS) cell genes was necessary for inducing the expression of immature status-related proteins, including Nanog, Ssea4, Tra-1-60, and Tra-1-80 in esophageal, stomach, colorectal, liver, pancreatic, and cholangiocellular cancer cells. Induced cells, but not parental cells, possessed the potential to express morphological patterns of ectoderm, mesoderm, and endoderm, which was supported by epigenetic studies, indicating methylation of DNA strands and the histone H3 protein at lysine 4 in promoter regions of pluripotency-associated genes such as NANOG. In in vitro analysis induced cells showed slow proliferation and were sensitized to differentiation-inducing treatment, and in vivo tumorigenesis was reduced in NOD/SCID mice. This study demonstrated that pluripotency was manifested in induced cells, and that the induced pluripotent cancer (iPC) cells were distinct from natural cancer cells with regard to their sensitivity to differentiation-inducing treatment. Retroviral-mediated introduction of iPC cells confers higher sensitivity to chemotherapeutic agents and differentiation-inducing treatment. PMID- 20018688 TI - Autoreactivity in an HIV-1 broadly reactive neutralizing antibody variable region heavy chain induces immunologic tolerance. AB - We previously reported that some of the rare broadly reactive, HIV-1 neutralizing antibodies are polyreactive, leading to the hypothesis that induction of these types of neutralizing antibody may be limited by immunologic tolerance. However, the notion that such antibodies are sufficiently autoreactive to trigger B cell tolerance is controversial. To test directly whether rare neutralizing HIV-1 antibodies can activate immunologic tolerance mechanisms, we generated a knock-in mouse in which the Ig heavy chain (HC) variable region rearrangement (V(H)DJ(H)) from the polyreactive and broadly neutralizing human monoclonal antibody 2F5 was targeted into the mouse Igh locus. In vitro, this insertion resulted in chimeric human/mouse 2F5 antibodies that were functionally similar to the human 2F5 antibody, including comparable reactivity to human and murine self-antigens. In vivo, the 2F5 V(H)DJ(H) insertion supported development of large- and small pre-B cells that expressed the chimeric human/mouse Igmu chain but not the production of immature B cells expressing membrane IgM. The developmental arrest exhibited in 2F5 V(H)DJ(H) knock-in mice is characteristic of other knock-in strains that express the Ig HC variable region of autoreactive antibodies and is consistent with the loss of immature B cells bearing 2F5 chimeric antibodies to central tolerance mechanisms. Moreover, homozygous 2F5 V(H)DJ(H) knock-in mice support reduced numbers of residual splenic B cells with low surface IgM density, severely diminished serum IgM levels, but normal to elevated quantities of serum IgGs that did not react with autoantigens. These features are consistent with elimination of 2F5 HC autoreactivity by additional negative selection mechanism(s) in the periphery. PMID- 20018689 TI - Polycomb Repressive Complex 2 and Trithorax modulate Drosophila longevity and stress resistance. AB - Polycomb Group (PcG) and Trithorax Group (TrxG) proteins are key epigenetic regulators of global transcription programs. Their antagonistic chromatin modifying activities modulate the expression of many genes and affect many biological processes. Here we report that heterozygous mutations in two core subunits of Polycomb Repressive Complex 2 (PRC2), the histone H3 lysine 27 (H3K27)-specific methyltransferase E(Z) and its partner, the H3 binding protein ESC, increase longevity and reduce adult levels of trimethylated H3K27 (H3K27me3). Mutations in trithorax (trx), a well known antagonist of Polycomb silencing, elevate the H3K27me3 level of E(z) mutants and suppress their increased longevity. Like many long-lived mutants, E(z) and esc mutants exhibit increased resistance to oxidative stress and starvation, and these phenotypes are also suppressed by trx mutations. This suppression strongly suggests that both the longevity and stress resistance phenotypes of PRC2 mutants are specifically due to their reduced levels of H3K27me3 and the consequent perturbation of Polycomb silencing. Consistent with this, long-lived E(z) mutants exhibit derepression of Abd-B, a well-characterized direct target of Polycomb silencing, and Odc1, a putative direct target implicated in stress resistance. These findings establish a role for PRC2 and TRX in the modulation of organismal longevity and stress resistance and indicate that moderate perturbation of Polycomb silencing can increase longevity. PMID- 20018691 TI - Evidence disputing deforestation as the cause for the collapse of the ancient Maya polity of Copan, Honduras. AB - Archaeologists have proposed diverse hypotheses to explain the collapse of the southern Maya lowland cities between the 8th and 10th centuries A.D. Although it generally is believed that no single factor was responsible, a commonly accepted cause is environmental degradation as a product of large-scale deforestation. To date, the most compelling scientific evidence used to support this hypothesis comes from the archaeological site of Copan, Honduras, where the analysis of a sediment core suggested a dramatic increase in forest clearance in the Late Classic period (A.D. 600-900). By contrast, in the work presented here, the authors' analysis of a longer sediment core demonstrates that forest cover increased from A.D. 400 to A.D. 900, with arboreal pollen accounting for 59.8 71.0% of the pollen assemblage by approximately A.D. 780-980. The highest levels of deforestation are found about 900 B.C. when, at its peak, herb pollen made up 89.8% of the assemblage. A second, although less pronounced, period of elevated deforestation peaked at approximately A.D. 400 when herb pollen reached 65.3% of the assemblage. The first deforestation event likely coincided with the widespread adoption of agriculture, a pattern found elsewhere in Mesoamerica. The second period of forest clearance probably was associated with the incursion of Maya speakers into the Copan Valley and their subsequent construction of the earliest levels of the Copan Acropolis. These results refute the former hypothesis that the ancient Maya responded to their increasingly large urban population by exhausting, rather than conserving, natural resources. PMID- 20018690 TI - Effective collaboration between marginal metallophilic macrophages and CD8+ dendritic cells in the generation of cytotoxic T cells. AB - The spleen is the lymphoid organ that induces immune responses toward blood-borne pathogens. Specialized macrophages in the splenic marginal zone are strategically positioned to phagocytose pathogens and cell debris, but are not known to play a role in the activation of T-cell responses. Here we demonstrate that splenic marginal metallophilic macrophages (MMM) are essential for cross-presentation of blood-borne antigens by splenic dendritic cells (DCs). Our data demonstrate that antigens targeted to MMM as well as blood-borne adenoviruses are efficiently captured by MMM and exclusively transferred to splenic CD8(+) DCs for cross presentation and for the activation of cytotoxic T lymphocytes. Depletion of macrophages in the marginal zone prevents cytotoxic T-lymphocyte activation by CD8(+) DCs after antibody targeting or adenovirus infection. Moreover, we show that tumor antigen targeting to MMM is very effective as antitumor immunotherapy. Our studies point to an important role for splenic MMM in the initial steps of CD8(+) T-cell immunity by capturing and concentrating blood-borne antigens and the transfer to cross-presenting DCs which can be used to design vaccination strategies to induce antitumor cytotoxic T-cell immunity. PMID- 20018692 TI - A tetrahedral entropy for water. AB - We introduce the space-dependent correlation function C (Q)(r) and time-dependent autocorrelation function C (Q)(t) of the local tetrahedral order parameter Q identical with Q(r,t). By using computer simulations of 512 waterlike particles interacting through the transferable interaction potential with five points (TIP5 potential), we investigate C (Q)(r) in a broad region of the phase diagram. We find that at low temperatures C (Q)(t) exhibits a two-step time-dependent decay similar to the self-intermediate scattering function and that the corresponding correlation time tau(Q) displays a dynamic cross-over from non-Arrhenius behavior for T > T (W) to Arrhenius behavior for T < T (W), where T (W) denotes the Widom temperature where the correlation length has a maximum as T is decreased along a constant-pressure path. We define a tetrahedral entropy S (Q) associated with the local tetrahedral order of water molecules and find that it produces a major contribution to the specific heat maximum at the Widom line. Finally, we show that tau(Q) can be extracted from S (Q) by using an analog of the Adam-Gibbs relation. PMID- 20018693 TI - DNA-DNA interactions in bacteriophage capsids are responsible for the observed DNA knotting. AB - Recent experiments showed that the linear double-stranded DNA in bacteriophage capsids is both highly knotted and neatly structured. What is the physical basis of this organization? Here we show evidence from stochastic simulation techniques that suggests that a key element is the tendency of contacting DNA strands to order, as in cholesteric liquid crystals. This interaction favors their preferential juxtaposition at a small twist angle, thus promoting an approximately nematic (and apolar) local order. The ordering effect dramatically impacts the geometry and topology of DNA inside phages. Accounting for this local potential allows us to reproduce the main experimental data on DNA organization in phages, including the cryo-EM observations and detailed features of the spectrum of DNA knots formed inside viral capsids. The DNA knots we observe are strongly delocalized and, intriguingly, this is shown not to interfere with genome ejection out of the phage. PMID- 20018694 TI - Red blood cell-mimicking synthetic biomaterial particles. AB - Biomaterials form the basis of current and future biomedical technologies. They are routinely used to design therapeutic carriers, such as nanoparticles, for applications in drug delivery. Current strategies for synthesizing drug delivery carriers are based either on discovery of materials or development of fabrication methods. While synthetic carriers have brought upon numerous advances in drug delivery, they fail to match the sophistication exhibited by innate biological entities. In particular, red blood cells (RBCs), the most ubiquitous cell type in the human blood, constitute highly specialized entities with unique shape, size, mechanical flexibility, and material composition, all of which are optimized for extraordinary biological performance. Inspired by this natural example, we synthesized particles that mimic the key structural and functional features of RBCs. Similar to their natural counterparts, RBC-mimicking particles described here possess the ability to carry oxygen and flow through capillaries smaller than their own diameter. Further, they can also encapsulate drugs and imaging agents. These particles provide a paradigm for the design of drug delivery and imaging carriers, because they combine the functionality of natural RBCs with the broad applicability and versatility of synthetic drug delivery particles. PMID- 20018695 TI - A feeling for the numbers in biology. AB - Although the quantitative description of biological systems has been going on for centuries, recent advances in the measurement of phenomena ranging from metabolism to gene expression to signal transduction have resulted in a new emphasis on biological numeracy. This article describes the confluence of two different approaches to biological numbers. First, an impressive array of quantitative measurements make it possible to develop intuition about biological numbers ranging from how many gigatons of atmospheric carbon are fixed every year in the process of photosynthesis to the number of membrane transporters needed to provide sugars to rapidly dividing Escherichia coli cells. As a result of the vast array of such quantitative data, the BioNumbers web site has recently been developed as a repository for biology by the numbers. Second, a complementary and powerful tradition of numerical estimates familiar from the physical sciences and canonized in the so-called "Fermi problems" calls for efforts to estimate key biological quantities on the basis of a few foundational facts and simple ideas from physics and chemistry. In this article, we describe these two approaches and illustrate their synergism in several particularly appealing case studies. These case studies reveal the impact that an emphasis on numbers can have on important biological questions. PMID- 20018696 TI - Context-dependent interaction leads to emergent search behavior in social aggregates. AB - Locating the source of an advected chemical signal is a common challenge facing many living organisms. When the advecting medium is characterized by either high Reynolds number or high Peclet number, the task becomes highly nontrivial due to the generation of heterogeneous, dynamically changing filamental concentrations that do not decrease monotonically with distance to the source. Defining search strategies that are effective in these environments has important implications for the understanding of animal behavior and for the design of biologically inspired technology. Here we present a strategy that is able to solve this task without the higher intelligence required to assess spatial gradient direction, measure the diffusive properties of the flow field, or perform complex calculations. Instead, our method is based on the collective behavior of autonomous individuals following simple social interaction rules which are modified according to the local conditions they are experiencing. Through these context-dependent interactions, the group is able to locate the source of a chemical signal and in doing so displays an awareness of the environment not present at the individual level. This behavior illustrates an alternative pathway to the evolution of higher cognitive capacity via the emergent, group-level intelligence that can result from local interactions. PMID- 20018697 TI - Multiscale mobility networks and the spatial spreading of infectious diseases. AB - Among the realistic ingredients to be considered in the computational modeling of infectious diseases, human mobility represents a crucial challenge both on the theoretical side and in view of the limited availability of empirical data. To study the interplay between short-scale commuting flows and long-range airline traffic in shaping the spatiotemporal pattern of a global epidemic we (i) analyze mobility data from 29 countries around the world and find a gravity model able to provide a global description of commuting patterns up to 300 kms and (ii) integrate in a worldwide-structured metapopulation epidemic model a timescale separation technique for evaluating the force of infection due to multiscale mobility processes in the disease dynamics. Commuting flows are found, on average, to be one order of magnitude larger than airline flows. However, their introduction into the worldwide model shows that the large-scale pattern of the simulated epidemic exhibits only small variations with respect to the baseline case where only airline traffic is considered. The presence of short-range mobility increases, however, the synchronization of subpopulations in close proximity and affects the epidemic behavior at the periphery of the airline transportation infrastructure. The present approach outlines the possibility for the definition of layered computational approaches where different modeling assumptions and granularities can be used consistently in a unifying multiscale framework. PMID- 20018698 TI - PGC-1alpha negatively regulates hepatic FGF21 expression by modulating the heme/Rev-Erb(alpha) axis. AB - FGF21 is a hormone produced in liver and fat that dramatically improves peripheral insulin sensitivity and lipid metabolism. We show here that obese mice with genetically reduced levels of a key hepatic transcriptional coactivator, PGC 1alpha, have improved whole-body insulin sensitivity with increased levels of hepatic and circulating FGF21. Gain- and loss-of-function studies in primary mouse hepatocytes show that hepatic FGF21 levels are regulated by the expression of PGC-1alpha. Importantly, PGC-1alpha-mediated reduction of FGF21 expression is dependent on Rev-Erbalpha and the expression of ALAS-1. ALAS-1 is a PGC-1alpha target gene and the rate-limiting enzyme in the synthesis of heme, a ligand for Rev-Erbalpha. Modulation of intracellular heme levels mimics the effect of PGC 1alpha on FGF21 expression, and inhibition of heme biosynthesis completely abrogates the down-regulation of FGF21 in response to PGC-1alpha. Thus, PGC 1alpha can impact hepatic and systemic metabolism by regulating the levels of a nuclear receptor ligand. PMID- 20018699 TI - Oscillations in supercoiling drive circadian gene expression in cyanobacteria. AB - The cyanobacterium Synechococcus elongatus PCC 7942 exhibits oscillations in mRNA transcript abundance with 24-h periodicity under continuous light conditions. The mechanism underlying these oscillations remains elusive--neither cis nor trans factors controlling circadian gene expression phase have been identified. Here, we show that the topological status of the chromosome is highly correlated with circadian gene expression state. We also demonstrate that DNA sequence characteristics of genes that appear monotonically activated and monotonically repressed by chromosomal relaxation during the circadian cycle are similar to those of supercoiling-responsive genes in Escherichia coli. Furthermore, perturbation of superhelical status within the physiological range elicits global changes in gene expression similar to those that occur during the normal circadian cycle. PMID- 20018700 TI - Nucleosome positioning by genomic excluding-energy barriers. AB - Recent genome-wide nucleosome mappings along with bioinformatics studies have confirmed that the DNA sequence plays a more important role in the collective organization of nucleosomes in vivo than previously thought. Yet in living cells, this organization also results from the action of various external factors like DNA-binding proteins and chromatin remodelers. To decipher the code for intrinsic chromatin organization, there is thus a need for in vitro experiments to bridge the gap between computational models of nucleosome sequence preferences and in vivo nucleosome occupancy data. Here we combine atomic force microscopy in liquid and theoretical modeling to demonstrate that a major sequence signaling in vivo are high-energy barriers that locally inhibit nucleosome formation rather than favorable positioning motifs. We show that these genomic excluding-energy barriers condition the collective assembly of neighboring nucleosomes consistently with equilibrium statistical ordering principles. The analysis of two gene promoter regions in Saccharomyces cerevisiae and the human genome indicates that these genomic barriers direct the intrinsic nucleosome occupancy of regulatory sites, thereby contributing to gene expression regulation. PMID- 20018701 TI - Identification and characterization of Bph14, a gene conferring resistance to brown planthopper in rice. AB - Planthoppers are highly destructive pests in crop production worldwide. Brown planthopper (BPH) causes the most serious damage of the rice crop globally among all rice pests. Growing resistant varieties is the most effective and environment friendly strategy for protecting the crop from BPH. More than 19 BPH-resistance genes have been reported and used to various extents in rice breeding and production. In this study, we cloned Bph14, a gene conferring resistance to BPH at seedling and maturity stages of the rice plant, using a map-base cloning approach. We show that Bph14 encodes a coiled-coil, nucleotide-binding, and leucine-rich repeat (CC-NB-LRR) protein. Sequence comparison indicates that Bph14 carries a unique LRR domain that might function in recognition of the BPH insect invasion and activating the defense response. Bph14 is predominantly expressed in vascular bundles, the site of BPH feeding. Expression of Bph14 activates the salicylic acid signaling pathway and induces callose deposition in phloem cells and trypsin inhibitor production after planthopper infestation, thus reducing the feeding, growth rate, and longevity of the BPH insects. Our work provides insights into the molecular mechanisms of rice defense against insects and facilitates the development of resistant varieties to control this devastating insect. PMID- 20018702 TI - Temporal latitudinal-gradient dynamics and tropical instability of deep-sea species diversity. AB - A benthic microfaunal record from the equatorial Atlantic Ocean over the past four glacial-interglacial cycles was investigated to understand temporal dynamics of deep-sea latitudinal species diversity gradients (LSDGs). The results demonstrate unexpected instability and high amplitude fluctuations of species diversity in the tropical deep ocean that are correlated with orbital-scale oscillations in global climate: Species diversity is low during glacial and high during interglacial periods. This implies that climate severely influences deep sea diversity, even at tropical latitudes, and that deep-sea LSDGs, while generally present for the last 36 million years, were weakened or absent during glacial periods. Temporally dynamic LSDGs and unstable tropical diversity require reconsideration of current ecological hypotheses about the generation and maintenance of biodiversity as they apply to the deep sea, and underscore the potential vulnerability and conservation importance of tropical deep-sea ecosystems. PMID- 20018703 TI - Dune formation under bimodal winds. AB - The study of dune morphology represents a valuable tool in the investigation of planetary wind systems--the primary factor controlling the dune shape is the wind directionality. However, our understanding of dune formation is still limited to the simplest situation of unidirectional winds: There is no model that solves the equations of sand transport under the most common situation of seasonally varying wind directions. Here we present the calculation of sand transport under bimodal winds using a dune model that is extended to account for more than one wind direction. Our calculations show that dunes align longitudinally to the resultant wind trend if the angle(w) between the wind directions is larger than 90 degrees. Under high sand availability, linear seif dunes are obtained, the intriguing meandering shape of which is found to be controlled by the dune height and by the time the wind lasts at each one of the two wind directions. Unusual dune shapes including the "wedge dunes" observed on Mars appear within a wide spectrum of bimodal dune morphologies under low sand availability. PMID- 20018704 TI - A unifying model for timing of walking onset in humans and other mammals. AB - The onset of walking is a fundamental milestone in motor development of humans and other mammals, yet little is known about what factors determine its timing. Hoofed animals start walking within hours after birth, rodents and small carnivores require days or weeks, and nonhuman primates take months and humans approximately a year to achieve this locomotor skill. Here we show that a key to the explanation for these differences is that time to the onset of walking counts from conception and not from birth, indicating that mechanisms underlying motor development constitute a functional continuum from pre- to postnatal life. In a multiple-regression model encompassing 24 species representative of 11 extant orders of placental mammals that habitually walk on the ground, including humans, adult brain mass accounted for 94% of variance in time to walking onset postconception. A dichotomous variable reflecting species differences in functional limb anatomy accounted for another 3.8% of variance. The model predicted the timing of walking onset in humans with high accuracy, showing that this milestone in human motor development occurs no later than expected given the mass of the adult human brain, which in turn reflects the duration of its ontogenetic development. The timing of motor development appears to be highly conserved in mammalian evolution as the ancestors of some of the species in the sample presented here diverged in phylogenesis as long as 100 million years ago. Fundamental patterns of early human life history may therefore have evolved before the evolution of primates. PMID- 20018705 TI - Missing and spurious interactions and the reconstruction of complex networks. AB - Network analysis is currently used in a myriad of contexts, from identifying potential drug targets to predicting the spread of epidemics and designing vaccination strategies and from finding friends to uncovering criminal activity. Despite the promise of the network approach, the reliability of network data is a source of great concern in all fields where complex networks are studied. Here, we present a general mathematical and computational framework to deal with the problem of data reliability in complex networks. In particular, we are able to reliably identify both missing and spurious interactions in noisy network observations. Remarkably, our approach also enables us to obtain, from those noisy observations, network reconstructions that yield estimates of the true network properties that are more accurate than those provided by the observations themselves. Our approach has the potential to guide experiments, to better characterize network data sets, and to drive new discoveries. PMID- 20018706 TI - Overfishing reduces resilience of kelp beds to climate-driven catastrophic phase shift. AB - A key consideration in assessing impacts of climate change is the possibility of synergistic effects with other human-induced stressors. In the ocean realm, climate change and overfishing pose two of the greatest challenges to the structure and functioning of marine ecosystems. In eastern Tasmania, temperate coastal waters are warming at approximately four times the global ocean warming average, representing the fastest rate of warming in the Southern Hemisphere. This has driven range extension of the ecologically important long-spined sea urchin (Centrostephanus rodgersii), which has now commenced catastrophic overgrazing of productive Tasmanian kelp beds leading to loss of biodiversity and important rocky reef ecosystem services. Coincident with the overgrazing is heavy fishing of reef-based predators including the spiny lobster Jasus edwardsii. By conducting experiments inside and outside Marine Protected Areas we show that fishing, by removing large predatory lobsters, has reduced the resilience of kelp beds against the climate-driven threat of the sea urchin and thus increased risk of catastrophic shift to widespread sea urchin barrens. This shows that interactions between multiple human-induced stressors can exacerbate nonlinear responses of ecosystems to climate change and limit the adaptive capacity of these systems. Management actions focused on reducing the risk of catastrophic phase shift in ecosystems are particularly urgent in the face of ongoing warming and unprecedented levels of predator removal from the world's oceans. PMID- 20018707 TI - Scanned-probe detection of electron spin resonance from a nitroxide spin probe. AB - We report an approach that extends the applicability of ultrasensitive force gradient detection of magnetic resonance to samples with spin-lattice relaxation times (T (1)) as short as a single cantilever period. To demonstrate the generality of the approach, which relies on detecting either cantilever frequency or phase, we used it to detect electron spin resonance from a T (1) = 1 ms nitroxide spin probe in a thin film at 4.2 K and 0.6 T. By using a custom fabricated cantilever with a 4 microm-diameter nickel tip, we achieve a magnetic resonance sensitivity of 400 Bohr magnetons in a 1 Hz bandwidth. A theory is presented that quantitatively predicts both the lineshape and the magnitude of the observed cantilever frequency shift as a function of field and cantilever sample separation. Good agreement was found between nitroxide T (1) 's measured mechanically and inductively, indicating that the cantilever magnet is not an appreciable source of spin-lattice relaxation here. We suggest that the new approach has a number of advantages that make it well suited to push magnetic resonance detection and imaging of nitroxide spin labels in an individual macromolecule to single-spin sensitivity. PMID- 20018708 TI - Accumulation of a differentiation regulator specifies transit amplifying division number in an adult stem cell lineage. AB - A key feature of many adult stem cell lineages is that stem cell daughters destined for differentiation undergo several transit amplifying (TA) divisions before initiating terminal differentiation, allowing few and infrequently dividing stem cells to produce many differentiated progeny. Although the number of progenitor divisions profoundly affects tissue (re)generation, and failure to control these divisions may contribute to cancer, the mechanisms that limit TA proliferation are not well understood. Here, we use a model stem cell lineage, the Drosophila male germ line, to investigate the mechanism that counts the number of TA divisions. The Drosophila Bag of Marbles (Bam) protein is required for male germ cells to cease spermatogonial TA divisions and initiate spermatocyte differentiation [McKearin DM, et al. (1990) Genes Dev 4:2242-2251]. Contrary to models involving dilution of a differentiation repressor, our results suggest that the switch from proliferation to terminal differentiation is triggered by accumulation of Bam protein to a critical threshold in TA cells and that the number of TA divisions is set by the timing of Bam accumulation with respect to the rate of cell cycle progression. PMID- 20018709 TI - Performance level modulates adult age differences in brain activation during spatial working memory. AB - Working memory (WM) shows pronounced age-related decline. Functional magnetic resonance imaging (fMRI) studies have revealed age differences in task-related brain activation. Evidence based primarily on episodic memory studies suggests that brain activation patterns can be modulated by task difficulty in both younger and older adults. In most fMRI aging studies on WM, however, performance level has not been considered, so that age differences in activation patterns are confounded with age differences in performance level. Here, we address this issue by comparing younger and older low and high performers in an event-related fMRI study. Thirty younger (20-30 years) and 30 older (60-70 years) healthy adults were tested with a spatial WM task with three load levels. A region-of-interest analysis revealed marked differences in the activation patterns between high and low performers in both age groups. Critically, among the older adults, a more "youth-like" load-dependent modulation of the blood oxygen level-dependent signal was associated with higher levels of spatial WM performance. These findings underscore the need of taking performance level into account when studying changes in functional brain activation patterns from early to late adulthood. PMID- 20018710 TI - Short paths in protein structure space originate in graph structure. PMID- 20018711 TI - Bandit solutions provide unified ethical models for randomized clinical trials and comparative effectiveness research. AB - As electronic medical records enable increasingly ambitious studies of treatment outcomes, ethical issues previously important only to limited clinical trials become relevant to unlimited whole populations. For randomized clinical trials, adaptive assignment strategies are known to expose substantially fewer patients to avoidable treatment failures than strategies with fixed assignments (e.g., equal sample sizes). An idealized adaptive case--the two-armed Bernoulli bandit problem--can be exactly optimized for a variety of ethically motivated cost functions that embody principles of duty-to-patient, but the solutions have been thought computationally infeasible when the numbers of patients in the study (the "horizon") is large. We report numerical experiments that yield a heuristic approximation that applies even to very large horizons, and we propose a near optimal strategy that remains valid even when the horizon is unknown or unbounded, thus applicable to comparative effectiveness studies on large populations or to standard-of-care recommendations. For the case in which the economic cost of treatment is a parameter, we give a heuristic, near-optimal strategy for determining the superior treatment (whether more or less costly) while minimizing resources wasted on any inferior, more expensive, treatment. Key features of our heuristics can be generalized to more complicated protocols. PMID- 20018712 TI - The N-terminus of histone H3 is required for de novo DNA methylation in chromatin. AB - DNA methylation and histone modification are two major epigenetic pathways that interplay to regulate transcriptional activity and other genome functions. Dnmt3L is a regulatory factor for the de novo DNA methyltransferases Dnmt3a and Dnmt3b. Although recent biochemical studies have revealed that Dnmt3L binds to the tail of histone H3 with unmethylated lysine 4 in vitro, the requirement of chromatin components for DNA methylation has not been examined, and functional evidence for the connection of histone tails to DNA methylation is still lacking. Here, we used the budding yeast Saccharomyces cerevisiae as a model system to investigate the chromatin determinants of DNA methylation through ectopic expression of murine Dnmt3a and Dnmt3L. We found that the N terminus of histone H3 tail is required for de novo methylation, while the central part encompassing lysines 9 and 27, as well as the H4 tail are dispensable. DNA methylation occurs predominantly in heterochromatin regions lacking H3K4 methylation. In mutant strains depleted of H3K4 methylation, the DNA methylation level increased 5-fold. The methylation activity of Dnmt3a largely depends on the Dnmt3L's PHD domain recognizing the histone H3 tail with unmethylated lysine 4. Functional analysis of Dnmt3L in mouse ES cells confirmed that the chromatin-recognition ability of Dnmt3L's PHD domain is indeed required for efficient methylation at the promoter of the endogenous Dnmt3L gene. These findings establish the N terminus of histone H3 tail with an unmethylated lysine 4 as a chromatin determinant for DNA methylation. PMID- 20018714 TI - Fast, background-free, 3D super-resolution optical fluctuation imaging (SOFI). AB - Super-resolution optical microscopy is a rapidly evolving area of fluorescence microscopy with a tremendous potential for impacting many fields of science. Several super-resolution methods have been developed over the last decade, all capable of overcoming the fundamental diffraction limit of light. We present here an approach for obtaining subdiffraction limit optical resolution in all three dimensions. This method relies on higher-order statistical analysis of temporal fluctuations (caused by fluorescence blinking/intermittency) recorded in a sequence of images (movie). We demonstrate a 5-fold improvement in spatial resolution by using a conventional wide-field microscope. This resolution enhancement is achieved in iterative discrete steps, which in turn allows the evaluation of images at different resolution levels. Even at the lowest level of resolution enhancement, our method features significant background reduction and thus contrast enhancement and is demonstrated on quantum dot-labeled microtubules of fibroblast cells. PMID- 20018715 TI - Hybridization kinetics is different inside cells. AB - It is generally expected that the kinetics of reactions inside living cells differs from the situation in bulk solutions. Macromolecular crowding and specific binding interactions could change the diffusion properties and the availability of free molecules. Their impact on reaction kinetics in the relevant context of living cells is still elusive, mainly because the difficulty of capturing fast kinetics in vivo. This article shows spatially resolved measurements of DNA hybridization kinetics in single living cells. HeLa cells were transfected with a FRET-labeled dsDNA probe by lipofection. We characterized the hybridization reaction kinetics with a kinetic range of 10 micros to 1 s by a combination of laser-driven temperature oscillations and stroboscopic fluorescence imaging. The time constant of the hybridization depended on DNA concentration within individual cells and between cells. A quantitative analysis of the concentration dependence revealed several-fold accelerated kinetics as compared with free solution for a 16-bp probe and decelerated kinetics for a 12 bp probe. We did not find significant effects of crowding agents on the hybridization kinetics in vitro. Our results suggest that the reaction rates in vivo are specifically modulated by binding interactions for the two probes, possibly triggered by their different lengths. In general, the presented imaging modality of temperature oscillation optical lock-in microscopy allows to probe biomolecular interactions in different cell compartments in living cells for systems biology. PMID- 20018716 TI - Navigating molecular worms inside chemical labyrinths. AB - Predicting whether a molecule can traverse chemical labyrinths of channels, tunnels, and buried cavities usually requires performing computationally intensive molecular dynamics simulations. Often one wants to screen molecules to identify ones that can pass through a given chemical labyrinth or screen chemical labyrinths to identify those that allow a given molecule to pass. Because it is impractical to test each molecule/labyrinth pair using computationally expensive methods, faster, approximate methods are used to prune possibilities, "triaging" the ability of a proposed molecule to pass through the given chemical labyrinth. Most pruning methods estimate chemical accessibility solely on geometry, treating atoms or groups of atoms as hard spheres with appropriate radii. Here, we explore geometric configurations for a moving "molecular worm," which replaces spherical probes and is assembled from solid blocks connected by flexible links. The key is to extend the fast marching method, which is an ordered upwind one-pass Dijkstra like method to compute optimal paths by efficiently solving an associated Eikonal equation for the cost function. First, we build a suitable cost function associated with each possible configuration, and second, we construct an algorithm that works in ensuing high-dimensional configuration space: at least seven dimensions are required to account for translational, rotational, and internal degrees of freedom. We demonstrate the algorithm to study shortest paths, compute accessible volume, and derive information on topology of the accessible part of a chemical labyrinth. As a model example, we consider an alkane molecule in a porous material, which is relevant to designing catalysts for oil processing. PMID- 20018717 TI - Prepubertal human spermatogonia and mouse gonocytes share conserved gene expression of germline stem cell regulatory molecules. AB - In the human testis, beginning at approximately 2 months of age, gonocytes are replaced by adult dark (Ad) and pale (Ap) spermatogonia that make up the spermatogonial stem cell (SSC) pool. In mice, the SSC pool arises from gonocytes approximately 6 days after birth. During puberty in both species, complete spermatogenesis is established by cells that differentiate from SSCs. Essentially pure populations of prepubertal human spermatogonia and mouse gonocytes were selected from testis biopsies and validated by confirming the presence of specific marker proteins in cells. Stem cell potential of germ cells was demonstrated by transplantation to mouse testes, following which the cells migrated to the basement membrane of the seminiferous tubule and were maintained similar to SSCs. Differential gene expression profiles generated between germ cells and testis somatic cells demonstrated that expression of genes previously identified as SSC and spermatogonial-specific markers (e.g., zinc-finger and BTB domain containing 16, ZBTB16) was greatly elevated in both human spermatogonia and mouse gonocytes compared to somatic cells. Several genes were expressed at significantly higher levels in germ cells of both species. Most importantly, genes known to be essential for mouse SSC self-renewal (e.g., Ret proto-oncogene, Ret; GDNF-family receptor alpha1, Gfr alpha1; and B-cell CLL/lymphoma 6, member B, Bcl6b) were more highly expressed in both prepubertal human spermatogonia and mouse gonocytes than in somatic cells. The results indicate remarkable conservation of gene expression, notably for self-renewal genes, in these prepubertal germline cells between two species that diverged phylogenetically approximately 75 million years ago. PMID- 20018713 TI - Allelic diversities in rice starch biosynthesis lead to a diverse array of rice eating and cooking qualities. AB - More than half of the world's population uses rice as a source of carbon intake every day. Improving grain quality is thus essential to rice consumers. The three main properties that determine rice eating and cooking quality--amylose content, gel consistency, and gelatinization temperature--correlate with one another, but the underlying mechanism of these properties remains unclear. Through an association analysis approach, we found that genes related to starch synthesis cooperate with each other to form a fine regulating network that controls the eating and cooking quality and defines the correlation among these three properties. Genetic transformation results verified the association findings and also suggested the possibility of developing elite cultivars through modification with selected major and/or minor starch synthesis-related genes. PMID- 20018718 TI - Epigenetic inactivation of the Sotos overgrowth syndrome gene histone methyltransferase NSD1 in human neuroblastoma and glioma. AB - Sotos syndrome is an autosomal dominant condition characterized by overgrowth resulting in tall stature and macrocephaly, together with an increased risk of tumorigenesis. The disease is caused by loss-of-function mutations and deletions of the nuclear receptor SET domain containing protein-1 (NSD1) gene, which encodes a histone methyltransferase involved in chromatin regulation. However, despite its causal role in Sotos syndrome and the typical accelerated growth of these patients, little is known about the putative contribution of NSD1 to human sporadic malignancies. Here, we report that NSD1 function is abrogated in human neuroblastoma and glioma cells by transcriptional silencing associated with CpG island-promoter hypermethylation. We also demonstrate that the epigenetic inactivation of NSD1 in transformed cells leads to the specifically diminished methylation of the histone lysine residues H4-K20 and H3-K36. The described phenotype is also observed in Sotos syndrome patients with NSD1 genetic disruption. Expression microarray data from NSD1-depleted cells, followed by ChIP analysis, revealed that the oncogene MEIS1 is one of the main NSD1 targets in neuroblastoma. Furthermore, we show that the restoration of NSD1 expression induces tumor suppressor-like features, such as reduced colony formation density and inhibition of cellular growth. Screening a large collection of different tumor types revealed that NSD1 CpG island hypermethylation was a common event in neuroblastomas and gliomas. Most importantly, NSD1 hypermethylation was a predictor of poor outcome in high-risk neuroblastoma. These findings highlight the importance of NSD1 epigenetic inactivation in neuroblastoma and glioma that leads to a disrupted histone methylation landscape and might have a translational value as a prognostic marker. PMID- 20018720 TI - General principles of attraction and competitive attraction as revealed by large cage studies of moths responding to sex pheromone. AB - Knowledge of how insects are actually affected by sex pheromones deployed throughout a crop so as to disrupt mating has lacked a mechanistic framework sufficient for guiding optimization of this environmentally friendly pest-control tactic. Major hypotheses are competitive attraction, desensitization, and camouflage. Working with codling moths, Cydia pomonella, in field cages millions of times larger than laboratory test tubes and at substrate concentrations trillions of times less than those typical for enzymes, we nevertheless demonstrate that mating disruption sufficiently parallels enzyme (ligand) substrate interactions so as to justify adoption of conceptual and analytical tools of biochemical kinetics. By doing so, we prove that commercial dispensers of codling moth pheromone first competitively attract and then deactivate males probably for the remainder of a night. No evidence was found for camouflage. We generated and now validate simple algebraic equations for attraction and competitive attraction that will guide optimization and broaden implementation of behavioral manipulations of pests. This analysis system also offers a unique approach to quantifying animal foraging behaviors and could find applications across the natural and social sciences. PMID- 20018719 TI - Functionality of the voltage-gated proton channel truncated in S4. AB - The voltage sensor domain (VSD) is the key module for voltage sensing in voltage gated ion channels and voltage-sensing phosphatases. Structurally, both the VSD and the recently discovered voltage-gated proton channels (Hv channels) voltage sensor only protein (VSOP) and Hv1 contain four transmembrane segments. The fourth transmembrane segment (S4) of Hv channels contains three periodically aligned arginines (R1, R2, R3). It remains unknown where protons permeate or how voltage sensing is coupled to ion permeation in Hv channels. Here we report that Hv channels truncated just downstream of R2 in the S4 segment retain most channel properties. Two assays, site-directed cysteine-scanning using accessibility of maleimide-reagent as detected by Western blotting and insertion into dog pancreas microsomes, both showed that S4 inserts into the membrane, even if it is truncated between the R2 and R3 positions. These findings provide important clues to the molecular mechanism underlying voltage sensing and proton permeation in Hv channels. PMID- 20018721 TI - Mutant p53 drives metastasis and overcomes growth arrest/senescence in pancreatic cancer. AB - TP53 mutation occurs in 50-75% of human pancreatic ductal adenocarcinomas (PDAC) following an initiating activating mutation in the KRAS gene. These p53 mutations frequently result in expression of a stable protein, p53(R175H), rather than complete loss of protein expression. In this study we elucidate the functions of mutant p53 (Trp53(R172H)), compared to knockout p53 (Trp53(fl)), in a mouse model of PDAC. First we find that although Kras(G12D) is one of the major oncogenic drivers of PDAC, most Kras(G12D)-expressing pancreatic cells are selectively lost from the tissue, and those that remain form premalignant lesions. Loss, or mutation, of Trp53 allows retention of the Kras(G12D)-expressing cells and drives rapid progression of these premalignant lesions to PDAC. This progression is consistent with failed growth arrest and/or senescence of premalignant lesions, since a mutant of p53, p53(R172P), which can still induce p21 and cell cycle arrest, is resistant to PDAC formation. Second, we find that despite similar kinetics of primary tumor formation, mutant p53(R172H), as compared with genetic loss of p53, specifically promotes metastasis. Moreover, only mutant p53(R172H) expressing tumor cells exhibit invasive activity in an in vitro assay. Importantly, in human PDAC, p53 accumulation significantly correlates with lymph node metastasis. In summary, by using 'knock-in' mutations of Trp53 we have identified two critical acquired functions of a stably expressed mutant form of p53 that drive PDAC; first, an escape from Kras(G12D)-induced senescence/growth arrest and second, the promotion of metastasis. PMID- 20018722 TI - Impact of oral bisphenol A at reference doses on intestinal barrier function and sex differences after perinatal exposure in rats. AB - Bisphenol A (BPA), a chemical estrogen widely used in the food-packaging industry and baby bottles, is recovered in human fluids (0.1-10 nM). Recent studies have reported that BPA is hormonally active at low doses, emphasizing the debate of a risk for human health. Estrogen receptors are expressed in the colon, and although the major route of BPA exposure is food, the effects on gut have received no attention. We first examined the endocrine disrupting potency of BPA on colonic paracellular permeability (CPP), experimental colitis, and visceral sensitivity in ovariectomized rats orally exposed to 5 mg/kg/d BPA (i.e., the no observed adverse effect level), 50 microg/kg/d BPA (i.e., tolerable daily intake), or lower doses. BPA dose-dependently decreased basal CPP, with a half maximal inhibitory dose of 5.2 microg/kg/d, 10-fold below the tolerable daily intake. This correlated with an increase in epithelial tight junction sealing, also observed in Caco-2 cells exposed to 10 nM BPA. When ovariectomized rats were fed with BPA at the no observed adverse effect level, the severity of colitis was reduced, whereas the same dose increased pain sensitivity to colorectal stimuli. We then examined the impact of perinatal exposure to BPA on intestinal permeability and inflammatory response in the offspring. In female rats, but not in male rats, perinatal BPA evoked a decrease of CPP in adulthood, whereas the proinflammatory response of colonic mucosa was strengthened. This study first demonstrates that the xenoestrogen BPA at reference doses influences intestinal barrier function and gut nociception. Moreover, perinatal exposure promotes the development of severe inflammation in adult female offspring only. PMID- 20018723 TI - Real-time observation of the transition from transcription initiation to elongation of the RNA polymerase. AB - The transition from initiation to elongation of the RNA polymerase (RNAP) is an important stage of transcription that often limits the production of the full length RNA. Little is known about the RNAP transition kinetics and the steps that dictate the transition rate, because of the challenge in monitoring subpopulations of the transient and heterogeneous transcribing complexes in rapid and real time. Here, we have dissected the complete transcription initiation pathway of T7 RNAP by using kinetic modeling of RNA synthesis and by determining the initiation (IC) to elongation (EC) transition kinetics at each RNA polymerization step using single-molecule and stopped-flow FRET methods. We show that the conversion of IC to EC in T7 RNAP consensus promoter occurs only after 8 to 12-nt synthesis, and the 12-nt synthesis represents a critical juncture in the transcriptional initiation pathway when EC formation is most efficient. We show that the slow steps of transcription initiation, including DNA scrunching/RNAP-promoter rotational changes during 5- to 8-nt synthesis, not the major conformational changes, dictate the overall rate of EC formation in T7 RNAP and represent key steps that regulate the synthesis of full-length RNA. PMID- 20018724 TI - The electronic structure of the primary electron donor of reaction centers of purple bacteria at atomic resolution as observed by photo-CIDNP 13C NMR. AB - Composed of the two bacteriochlorophyll cofactors, P(L) and P(M), the special pair functions as the primary electron donor in bacterial reaction centers of purple bacteria of Rhodobacter sphaeroides. Under light absorption, an electron is transferred to a bacteriopheophytin and a radical pair is produced. The occurrence of the radical pair is linked to the production of enhanced nuclear polarization called photochemically induced dynamic nuclear polarization (photo CIDNP). This effect can be used to study the electronic structure of the special pair at atomic resolution by detection of the strongly enhanced nuclear polarization with laser-flash photo-CIDNP magic-angle spinning NMR on the carotenoid-less mutant R26. In the electronic ground state, P(L) is strongly disturbed, carrying a slightly negative charge. In the radical cation state, the ratio of total electron spin densities between P(L) and P(M) is 2:1, although it is 2.5:1 for the pyrrole carbons, 2.2:1 for all porphyrinic carbons, and 4:1 for the pyrrole nitrogen. It is shown that the symmetry break between the electronic structures in the electronic ground state and in the radical cation state is an intrinsic property of the special pair supermolecule, which is particularly attributable to a modification of the structure of P(L). The significant difference in electron density distribution between the ground and radical cation states is explained by an electric polarization effect of the nearby histidine. PMID- 20018725 TI - The helicase protein DHX29 promotes translation initiation, cell proliferation, and tumorigenesis. AB - Translational control plays an important role in cell growth and tumorigenesis. Cap-dependent translation initiation of mammalian mRNAs with structured 5'UTRs requires the DExH-box protein, DHX29, in vitro. Here we show that DHX29 is important for translation in vivo. Down-regulation of DHX29 leads to impaired translation, resulting in disassembly of polysomes and accumulation of mRNA-free 80S monomers. DHX29 depletion also impedes cancer cell growth in culture and in xenografts. Thus, DHX29 is a bona fide translation initiation factor that potentially can be exploited as a target to inhibit cancer cell growth. PMID- 20018726 TI - Social isolation dysregulates endocrine and behavioral stress while increasing malignant burden of spontaneous mammary tumors. AB - In a life span study, we examined how the social environment regulates naturally occurring tumor development and malignancy in genetically prone Sprague-Dawley rats. We randomly assigned this gregarious species to live either alone or in groups of five female rats. Mammary tumor burden among social isolates increased to 84 times that of age-matched controls, as did malignancy, specifically a 3.3 relative risk for ductal carcinoma in situ and invasive ductal carcinoma, the most common early breast cancers in women. Importantly, isolation did not extend ovarian function in late middle age; in fact, isolated animals were exposed to lower levels of estrogen and progesterone in the middle-age period of mammary tumor growth, with unchanged tumor estrogen and progesterone receptor status. Isolates, however, did develop significant dysregulation of corticosterone responses to everyday stressors manifest in young adulthood, months before tumor development, and persisting into old age. Among isolates, corticosterone response to an acute stressor was enhanced and recovery was markedly delayed, each associated with increased mammary tumor progression. In addition to being stressed and tumor prone, an array of behavioral measures demonstrated that socially isolated females possessed an anxious, fearful, and vigilant phenotype. Our model provides a framework for studying the interaction of social neglect with genetic risk to identify mechanisms whereby psychosocial stressors increase growth and malignancy of breast cancer. PMID- 20018727 TI - Transcriptional response in the peripheral blood of patients infected with Salmonella enterica serovar Typhi. AB - We used microarrays and transcriptional profiling of peripheral blood to investigate the host response of 29 individuals who contracted typhoid fever in the Mekong Delta region of Vietnam. Samples were taken over a nine month period encompassing acute disease, convalescence, and recovery. We found that typhoid fever induced a distinct and highly reproducible signature in the peripheral blood that changed during treatment and convalescence, returning in the majority of cases to the "normal" profile as measured in healthy uninfected controls. Unexpectedly, there was a strong, distinct signature of convalescence present at day 9 after infection that remained virtually unchanged one month after acute infection and in some cases persisted as long as nine months despite a complete clinical recovery in all patients. Patients who retain the convalescent signature may be genetically or temporarily incapable of developing an effective immune response and may be more susceptible to reinfection, relapse, or the establishment of a carrier state. PMID- 20018728 TI - Planarian Hedgehog/Patched establishes anterior-posterior polarity by regulating Wnt signaling. AB - Despite long-standing interest, the molecular mechanisms underlying the establishment of anterior-posterior (AP) polarity remain among the unsolved mysteries in metazoans. In the planarians (a family of flatworms), canonical Wnt/beta-catenin signaling is required for posterior specification, as it is in many animals. However, the molecular mechanisms regulating the posterior-specific induction of Wnt genes according to the AP polarity have remained unclear. Here, we demonstrate that Hedgehog (Hh) signaling is responsible for the establishment of AP polarity via its regulation of the transcription of Wnt family genes during planarian regeneration. We found that RNAi gene knockdown of Dugesia japonica patched (Djptc) caused ectopic tail formation in the anterior blastema of body fragments, resulting in bipolar-tails regeneration. In contrast, RNAi of hedgehog (Djhh) and gli (Djgli) caused bipolar-heads regeneration. We show that Patched mediated Hh signaling was crucial for posterior specification, which is established by regulating the transcription of Wnt genes via downstream Gli activity. Moreover, differentiated cells were responsible for the posterior specification of undifferentiated stem cells through Wnt/beta-catenin signaling. Surprisingly, Djhh was expressed in neural cells all along the ventral nerve cords (along the AP axis), but not in the posterior blastema of body fragments, where the expression of Wnt genes was induced for posteriorization. We therefore propose that Hh signals direct head or tail regeneration according to the AP polarity, which is established by Hh signaling activity along the body's preexisting nervous system. PMID- 20018729 TI - Expression of mutant huntingtin in mouse brain astrocytes causes age-dependent neurological symptoms. AB - Huntington disease (HD) is an inherited neurological disorder caused by a polyglutamine expansion in the protein huntingtin and is characterized by selective neurodegeneration that preferentially occurs in striatal medium spiny neurons. Because the medium spiny neurons are innervated abundantly by glutamatergic axons from cortical neurons, the preferential degeneration in the striatal neurons supports the glutamate excitotoxicity theory for HD pathogenesis. Thus, glutamate uptake by glia may be particularly important for preventing glutamate excitotoxicity in HD. Although mutant huntingtin is expressed ubiquitously in various types of cells, it accumulates and forms aggregates in fewer glial cells than in neuronal cells. It remains largely unknown whether and how mutant huntingtin in glia can contribute to the neurological symptoms of HD. We generated transgenic mice that express N-terminal mutant huntingtin in astrocytes, a major type of glial cell that remove extracellular glutamate in the brain. Although transgenic mutant huntingtin in astrocytes is expressed below the endogenous level, it can cause age-dependent neurological phenotypes in transgenic mice. Mice expressing mutant huntingtin show body weight loss, have motor function deficits, and die earlier than wild type or control transgenic mice. We also found that mutant huntingtin in astrocytes decreases the expression of glutamate transporter by increasing its binding to Sp1 and reducing the association of Sp1 with the promoter of glutamate transporter. These results imply an important role for glial mutant huntingtin in HD pathology and suggest possibilities for treatment. PMID- 20018730 TI - AID-induced decrease in topoisomerase 1 induces DNA structural alteration and DNA cleavage for class switch recombination. AB - To initiate class switch recombination (CSR) activation-induced cytidine deaminase (AID) induces staggered nick cleavage in the S region, which lies 5' to each Ig constant region gene and is rich in palindromic sequences. Topoisomerase 1 (Top1) controls the supercoiling of DNA by nicking, rotating, and religating one strand of DNA. Curiously, Top1 reduction or AID overexpression causes the genomic instability. Here, we report that the inactivation of Top1 by its specific inhibitor camptothecin drastically blocked both the S region cleavage and CSR, indicating that Top1 is responsible for the S region cleavage in CSR. Surprisingly, AID expression suppressed Top1 mRNA translation and reduced its protein level. In addition, the decrease in the Top1 protein by RNA-mediated knockdown augmented the AID-dependent S region cleavage, as well as CSR. Furthermore, Top1 reduction altered DNA structure of the Smu region. Taken together, AID-induced Top1 reduction alters S region DNA structure probably to non-B form, on which Top1 can introduce nicks but cannot religate, resulting in S region cleavage. PMID- 20018731 TI - Random K-noncrossing RNA structures. AB - In this paper, we introduce a combinatorial framework that provides an interpretation of RNA pseudoknot structures as sampling paths of a Markov process. Our results facilitate a variety of applications ranging from the energy based sampling of pseudoknot structures as well as the ab initio folding via hidden Markov models. Our main result is an algorithm that generates RNA pseudoknot structures with uniform probability. This algorithm serves as a steppingstone to sequence-specific as well as energy-based transition probabilities. The approach employs a correspondence between pseudoknot structures, parametrized in terms of the maximal number of mutually crossing arcs and certain tableau sequences. The latter can be viewed as lattice paths. The main idea of this paper is to view each such lattice path as a sampling path of a stochastic process and to make use of D-finiteness for the efficient computation of the corresponding transition probabilities. PMID- 20018732 TI - Sigma-1 receptors regulate hippocampal dendritic spine formation via a free radical-sensitive mechanism involving Rac1xGTP pathway. AB - Sigma-1 receptors (Sig-1Rs) are endoplasmic reticulum (ER)-resident proteins known to be involved in learning and memory. Dendritic spines in hippocampal neurons play important roles in neuroplasticity and learning and memory. This study tested the hypothesis that Sig-1Rs might regulate denritic spine formation in hippocampal neurons and examined potential mechanisms therein. In rat hippocampal primary neurons, the knockdown of Sig-1Rs by siRNAs causes a deficit in the formation of dendritic spines that is unrelated to ER Ca(2+) signaling or apoptosis, but correlates with the mitochondrial permeability transition and cytochrome c release, followed by caspase-3 activation, Tiam1 cleavage, and a reduction in Rac1.GTP. Sig-1R-knockdown neurons contain higher levels of free radicals when compared to control neurons. The activation of superoxide dismutase or the application of the hydroxyl-free radical scavenger N-acetyl cysteine (NAC) to the Sig-1R-knockdown neurons rescues dendritic spines and mitochondria from the deficits caused by Sig-1R siRNA. Further, the caspase-3-resistant TIAM1 construct C1199DN, a stable guanine exchange factor able to constitutively activate Rac1 in the form of Rac1.GTP, also reverses the siRNA-induced dendritic spine deficits. In addition, constitutively active Rac1.GTP reverses this deficit. These results implicate Sig-1Rs as endogenous regulators of hippopcampal dendritic spine formation and suggest a free radical-sensitive ER-mitochondrion Rac1.GTP pathway in the regulation of dendritic spine formation in the hippocampus. PMID- 20018733 TI - Activation of EGFR on monocytes is required for human cytomegalovirus entry and mediates cellular motility. AB - Human cytomegalovirus (HCMV) rapidly induces a mobile and functionally unique proinflammatory monocyte following infection that is proposed to mediate viral spread. The cellular pathways used by HCMV to initiate these biological changes remain unknown. Here, we document the expression of the epidermal growth factor receptor (EGFR) on the surface of human peripheral blood monocytes but not on other blood leukocyte populations. Inhibition of EGFR signaling abrogated viral entry into monocytes, indicating that EGFR can serve as a cellular tropism receptor. Moreover, HCMV-activated EGFR was required for the induction of monocyte motility and transendothelial migration, two biological events required for monocyte extravasation into peripheral tissue, and thus viral spread. Transcriptome analysis revealed that HCMV-mediated EGFR signaling up-regulated neural Wiskott-Aldrich syndrome protein (N-WASP), an actin nucleator whose expression and function are normally limited in leukocytes. Knockdown of N-WASP expression blocked HCMV-induced but not phorbol 12-myristate 13-acetate (PMA) induced monocyte motility, suggesting that a switch to and/or the distinct use of a new actin nucleator controlling motility occurs during HCMV infection of monocytes. Together, these data provide evidence that EGFR plays an essential role in the immunopathobiology of HCMV by mediating viral entry into monocytes and stimulating the aberrant biological activity that promotes hematogenous dissemination. PMID- 20018735 TI - Temperature profile in the lowermost mantle from seismological and mineral physics joint modeling. AB - The internal structure of the core-mantle boundary (CMB) region of the Earth plays a crucial role in controlling the dynamics and evolution of our planet. We have developed a comprehensive model based on the radial variations of shear velocity in the D'' layer (the base of the lower mantle) and the high-P,T elastic properties of major candidate minerals, including the effects of post-perovskite phase changes. This modeling shows a temperature profile in the lowermost mantle with a CMB temperature of 3,800 +/- 200 K, which suggests that lateral temperature variations of 200-300 K can explain much of the large velocity heterogeneity observed in D''. A single-crossing phase transition model was found to be more favorable in reproducing the observed seismic wave velocity structure than a double-crossing phase transition model. PMID- 20018734 TI - Plasmodium falciparum var gene expression is modified by host immunity. AB - Plasmodium falciparum erythrocyte membrane protein 1 (PfEMP1) is a potentially important family of immune targets, which play a central role in the host parasite interaction by binding to various host molecules. They are encoded by a diverse family of genes called var, of which there are approximately 60 copies in each parasite genome. In sub-Saharan Africa, although P. falciparum infection occurs throughout life, severe malarial disease tends to occur only in childhood. This could potentially be explained if (i) PfEMP1 variants differ in their capacity to support pathogenesis of severe malaria and (ii) this capacity is linked to the likelihood of each molecule being recognized and cleared by naturally acquired antibodies. Here, in a study of 217 Kenyan children with malaria, we show that expression of a group of var genes "cys2," containing a distinct pattern of cysteine residues, is associated with low host immunity. Expression of cys2 genes was associated with parasites from young children, those with severe malaria, and those with a poorly developed antibody response to parasite-infected erythrocyte surface antigens. Cys-2 var genes form a minor component of all genomic var repertoires analyzed to date. Therefore, the results are compatible with the hypothesis that the genomic var gene repertoire is organized such that PfEMP1 molecules that confer the most virulence to the parasite tend also to be those that are most susceptible to the development of host immunity. This may help the parasite to adapt effectively to the development of host antibodies through modification of the host-parasite relationship. PMID- 20018736 TI - Structural determinants of ion permeation in CRAC channels. AB - CRAC channels generate Ca(2+) signals critical for the activation of immune cells and exhibit an intriguing pore profile distinguished by extremely high Ca(2+) selectivity, low Cs(+) permeability, and small unitary conductance. To identify the ion conduction pathway and gain insight into the structural bases of these permeation characteristics, we introduced cysteine residues in the CRAC channel pore subunit, Orai1, and probed their accessibility to various thiol-reactive reagents. Our results indicate that the architecture of the ion conduction pathway is characterized by a flexible outer vestibule formed by the TM1-TM2 loop, which leads to a narrow pore flanked by residues of a helical TM1 segment. Residues in TM3, and specifically, E190, a residue considered important for ion selectivity, are not close to the pore. Moreover, the outer vestibule does not significantly contribute to ion selectivity, implying that Ca(2+) selectivity is conferred mainly by E106. The ion conduction pathway is sufficiently narrow along much of its length to permit stable coordination of Cd(2+) by several TM1 residues, which likely explains the slow flux of ions within the restrained geometry of the pore. These results provide a structural framework to understand the unique permeation properties of CRAC channels. PMID- 20018737 TI - Gangliosides play pivotal roles in the regulation of complement systems and in the maintenance of integrity in nerve tissues. AB - Gangliosides are considered to be essential in the maintenance and repair of nervous tissues; however, the mechanisms for neurodegeneration caused by ganglioside defects are unknown. We examined gene expression profiles in double knockout (DKO) mice of GM2/GD2 synthase and GD3 synthase genes and showed that the majority of complement genes and their receptors were up-regulated in cerebellum in DKO mice. Inflammatory reactions were demonstrated in those tissues by measuring up-regulated inflammatory cytokines, indicating the presence of complement activation and inflammation as reported in Alzheimer's disease. Immunoblotting of fractionated membrane extracts by sucrose density gradient revealed that complement-regulatory molecules such as decay-accelerating factor and CD59 were dispersed from glycolipid-enriched microdomain/rafts in DKO cerebellum. Immunohistostaining of these molecules showed disordered membrane localization. These results suggested that dysfunction of complement-regulatory molecules may be due to abnormal glycolipid-enriched microdomain/rafts that triggered complement activation, subsequent inflammation, and neurodegeneration in DKO mice. Generation of the triple KO mice lacking complement activity in addition to the two glycosyltransferases suggested that complement activation is involved in the inflammatory reactions and neurodegeneration caused by the ganglioside deficiency. PMID- 20018738 TI - High-resolution protein complexes from integrating genomic information with molecular simulation. AB - Bacteria use two-component signal transduction systems (TCS) extensively to sense and react to external stimuli. In these, a membrane-bound sensor histidine kinase (SK) autophosphorylates in response to an environmental stimulus and transfers the phosphoryl group to a transcription factor/response regulator (RR) that mediates the cellular response. The complex between these two proteins is ruled by transient interactions, which provides a challenge to experimental structure determination techniques. The functional and structural homolog of an SK/RR pair Spo0B/Spo0F, however, has been structurally resolved. Here, we describe a method capable of generating structural models of such transient protein complexes. By using existing structures of the individual proteins, our method combines bioinformatically derived contact residue information with molecular dynamics simulations. We find crystal resolution accuracy with existing crystallographic data when reconstituting the known system Spo0B/Spo0F. Using this approach, we introduce a complex structure of TM0853/TM0468 as an exemplary SK/RR TCS, consistent with all experimentally available data. PMID- 20018739 TI - Probing ribosome-nascent chain complexes produced in vivo by NMR spectroscopy. AB - The means by which a polypeptide chain acquires its unique 3-D structure is a fundamental question in biology. During its synthesis on the ribosome, a nascent chain (NC) emerges vectorially and will begin to fold in a cotranslational fashion. The complex environment of the cell, coupled with the gradual emergence of the ribosome-tethered NC during its synthesis, imposes conformational restraints on its folding landscape that differ from those placed on an isolated protein when stimulated to fold following denaturation in solution. To begin to examine cotranslational folding as it would occur within a cell, we produce highly selective, isotopically labeled NCs bound to isotopically silent ribosomes in vivo. We then apply NMR spectroscopy to study, at a residue specific level, the conformation of NCs consisting of different fractional lengths of the polypeptide chain corresponding to a given protein. This combined approach provides a powerful means of generating a series of snapshots of the folding of the NC as it emerges from the ribosome. Application of this strategy to the NMR analysis of the progressive synthesis of an Ig-like domain reveals the existence of a partially folded ribosome-bound species that is likely to represent an intermediate species populated during the cotranslational folding process. PMID- 20018740 TI - Ancient DNA reveals late survival of mammoth and horse in interior Alaska. AB - Causes of late Quaternary extinctions of large mammals ("megafauna") continue to be debated, especially for continental losses, because spatial and temporal patterns of extinction are poorly known. Accurate latest appearance dates (LADs) for such taxa are critical for interpreting the process of extinction. The extinction of woolly mammoth and horse in northwestern North America is currently placed at 15,000-13,000 calendar years before present (yr BP), based on LADs from dating surveys of macrofossils (bones and teeth). Advantages of using macrofossils to estimate when a species became extinct are offset, however, by the improbability of finding and dating the remains of the last-surviving members of populations that were restricted in numbers or confined to refugia. Here we report an alternative approach to detect 'ghost ranges' of dwindling populations, based on recovery of ancient DNA from perennially frozen and securely dated sediments (sedaDNA). In such contexts, sedaDNA can reveal the molecular presence of species that appear absent in the macrofossil record. We show that woolly mammoth and horse persisted in interior Alaska until at least 10,500 yr BP, several thousands of years later than indicated from macrofossil surveys. These results contradict claims that Holocene survival of mammoths in Beringia was restricted to ecologically isolated high-latitude islands. More importantly, our finding that mammoth and horse overlapped with humans for several millennia in the region where people initially entered the Americas challenges theories that megafaunal extinction occurred within centuries of human arrival or were due to an extraterrestrial impact in the late Pleistocene. PMID- 20018741 TI - Ecology of the rare microbial biosphere of the Arctic Ocean. AB - Understanding the role of microbes in the oceans has focused on taxa that occur in high abundance; yet most of the marine microbial diversity is largely determined by a long tail of low-abundance taxa. This rare biosphere may have a cosmopolitan distribution because of high dispersal and low loss rates, and possibly represents a source of phylotypes that become abundant when environmental conditions change. However, the true ecological role of rare marine microorganisms is still not known. Here, we use pyrosequencing to describe the structure and composition of the rare biosphere and to test whether it represents cosmopolitan taxa or whether, similar to abundant phylotypes, the rare community has a biogeography. Our examination of 740,353 16S rRNA gene sequences from 32 bacterial and archaeal communities from various locations of the Arctic Ocean showed that rare phylotypes did not have a cosmopolitan distribution but, rather, followed patterns similar to those of the most abundant members of the community and of the entire community. The abundance distributions of rare and abundant phylotypes were different, following a log-series and log-normal model, respectively, and the taxonomic composition of the rare biosphere was similar to the composition of the abundant phylotypes. We conclude that the rare biosphere has a biogeography and that its tremendous diversity is most likely subjected to ecological processes such as selection, speciation, and extinction. PMID- 20018742 TI - Characterization of an electron conduit between bacteria and the extracellular environment. AB - A number of species of Gram-negative bacteria can use insoluble minerals of Fe(III) and Mn(IV) as extracellular respiratory electron acceptors. In some species of Shewanella, deca-heme electron transfer proteins lie at the extracellular face of the outer membrane (OM), where they can interact with insoluble substrates. To reduce extracellular substrates, these redox proteins must be charged by the inner membrane/periplasmic electron transfer system. Here, we present a spectro-potentiometric characterization of a trans-OM icosa-heme complex, MtrCAB, and demonstrate its capacity to move electrons across a lipid bilayer after incorporation into proteoliposomes. We also show that a stable MtrAB subcomplex can assemble in the absence of MtrC; an MtrBC subcomplex is not assembled in the absence of MtrA; and MtrA is only associated to the membrane in cells when MtrB is present. We propose a model for the modular organization of the MtrCAB complex in which MtrC is an extracellular element that mediates electron transfer to extracellular substrates and MtrB is a trans-OM spanning beta-barrel protein that serves as a sheath, within which MtrA and MtrC exchange electrons. We have identified the MtrAB module in a range of bacterial phyla, suggesting that it is widely used in electron exchange with the extracellular environment. PMID- 20018743 TI - Subnanometer atomic force microscopy of peptide-mineral interactions links clustering and competition to acceleration and catastrophe. AB - In vitro observations have revealed major effects on the structure, growth, and composition of biomineral phases, including stabilization of amorphous precursors, acceleration and inhibition of kinetics, and alteration of impurity signatures. However, deciphering the mechanistic sources of these effects has been problematic due to a lack of tools to resolve molecular structures on mineral surfaces during growth. Here we report atomic force microscopy investigations using a system designed to maximize resolution while minimizing contact force. By imaging the growth of calcium-oxalate monohydrate under the influence of aspartic-rich peptides at single-molecule resolution, we reveal how the unique interactions of polypeptides with mineral surfaces lead to acceleration, inhibition, and switching of growth between two distinct states. Interaction with the positively charged face of calcium-oxalate monohydrate leads to formation of a peptide film, but the slow adsorption kinetics and gradual relaxation to a well-bound state result in time-dependent effects. These include a positive feedback between peptide adsorption and step inhibition described by a mathematical catastrophe that results in growth hysteresis, characterized by rapid switching from fast to near-zero growth rates for very small reductions in supersaturation. Interactions with the negatively charged face result in formation of peptide clusters that impede step advancement. The result is a competition between accelerated solute attachment and inhibition due to blocking of the steps by the clusters. The findings have implications for control of pathological mineralization and suggest artificial strategies for directing crystallization. PMID- 20018744 TI - NMR analysis of G-protein betagamma subunit complexes reveals a dynamic G(alpha) Gbetagamma subunit interface and multiple protein recognition modes. AB - G-protein betagamma (Gbetagamma) subunits interact with a wide range of molecular partners including: G(alpha) subunits, effectors, peptides, and small molecule inhibitors. The molecular mechanisms underlying the ability to accommodate this wide range of structurally distinct binding partners are not well understood. To uncover the role of protein flexibility and alterations in protein conformation in molecular recognition by Gbetagamma, a method for site-specific (15)N-labeling of Gbeta-Trp residue backbone and indole amines in insect cells was developed. Transverse Relaxation Optimized Spectroscopy-Heteronuclear Single-Quantum Coherence Nuclear Magnetic Resonance (TROSY-HSQC NMR) analysis of (15)N-Trp Gbetagamma identified well-dispersed signals for the individual Trp residue side chain and amide positions. Surprisingly, a wide range of signal intensities was observed in the spectrum, likely representing a range of backbone and side chain mobilities. The signal for GbetaW99 indole was very intense, suggesting a high level of mobility on the protein surface and molecular dynamics simulations indicate that GbetaW99 is highly mobile on the nanosecond timescale in comparison with other Gbeta tryptophans. Binding of peptides and phosducin dramatically altered the mobility of GbetaW99 and GbetaW332 in the binding site and the chemical shifts at sites distant from the direct binding surface in distinct ways. In contrast, binding of G(alpha)(i1)-GDP to Gbetagamma had relatively little effect on the spectrum and, most surprisingly, did not significantly alter Trp mobility at the subunit interface. This suggests the inactive heterotrimer in solution adopts a conformation with an open subunit interface a large percentage of the time. Overall, these data show that Gbetagamma subunits explore a range of conformations that can be exploited during molecular recognition by diverse binding partners. PMID- 20018745 TI - Nanoparticles reveal that human cervicovaginal mucus is riddled with pores larger than viruses. AB - The mechanisms by which mucus helps prevent viruses from infecting mucosal surfaces are not well understood. We engineered non-mucoadhesive nanoparticles of various sizes and used them as probes to determine the spacing between mucin fibers (pore sizes) in fresh undiluted human cervicovaginal mucus (CVM) obtained from volunteers with healthy vaginal microflora. We found that most pores in CVM have diameters significantly larger than human viruses (average pore size 340 +/- 70 nm; range approximately 50-1800 nm). This mesh structure is substantially more open than the 15-100-nm spacing expected assuming mucus consists primarily of a random array of individual mucin fibers. Addition of a nonionic detergent to CVM caused the average pore size to decrease to 130 +/- 50 nm. This suggests hydrophobic interactions between lipid-coated "naked" protein regions on mucins normally cause mucin fibers to self-condense and/or bundle with other fibers, creating mucin "cables" at least three times thicker than individual mucin fibers. Although the native mesh structure is not tight enough to trap most viruses, we found that herpes simplex virus (approximately 180 nm) was strongly trapped in CVM, moving at least 8,000-fold slower than non-mucoadhesive 200-nm nanoparticles. This work provides an accurate measurement of the pore structure of fresh, hydrated ex vivo CVM and demonstrates that mucoadhesion, rather than steric obstruction, may be a critical protective mechanism against a major sexually transmitted virus and perhaps other viruses. PMID- 20018746 TI - NMR reveals pathway for ferric mineral precursors to the central cavity of ferritin. AB - Ferritin is a multimeric nanocage protein that directs the reversible biomineralization of iron. At the catalytic ferroxidase site two iron(II) ions react with dioxygen to form diferric species. In order to study the pathway of iron(III) from the ferroxidase site to the central cavity a new NMR strategy was developed to manage the investigation of a system composed of 24 monomers of 20 kDa each. The strategy is based on (13)C-(13)C solution NOESY experiments combined with solid-state proton-driven (13)C-(13)C spin diffusion and 3D coherence transfer experiments. In this way, 75% of amino acids were recognized and 35% sequence-specific assigned. Paramagnetic broadening, induced by iron(III) species in solution (13)C-(13)C NOESY spectra, localized the iron within each subunit and traced the progression to the central cavity. Eight iron ions fill the 20-A-long iron channel from the ferrous/dioxygen oxidoreductase site to the exit into the cavity, inside the four-helix bundle of each subunit, contrasting with short paths in models. Magnetic susceptibility data support the formation of ferric multimers in the iron channels. Multiple iron channel exits are near enough to facilitate high concentration of iron that can mineralize in the ferritin cavity, illustrating advantages of the multisubunit cage structure. PMID- 20018747 TI - Small-molecule tools for dissecting the roles of SSB/protein interactions in genome maintenance. AB - Bacterial single-stranded DNA-binding proteins (SSBs) help to recruit a diverse array of genome maintenance enzymes to their sites of action through direct protein interactions. For all cases examined to date, these interactions are mediated by the evolutionarily conserved C terminus of SSB (SSB-Ct). The essential nature of SSB protein interactions makes inhibitors that block SSB complex formation valuable biochemical tools and attractive potential antibacterial agents. Here, we identify four small molecules that disrupt complexes formed between Escherichia coli SSB and Exonuclease I (ExoI), a well studied SSB-interacting enzyme. Each compound disrupts ExoI/SSB-Ct peptide complexes and abrogates SSB stimulation of ExoI nuclease activity. Structural and biochemical studies support a model for three of the compounds in which they compete with SSB for binding to ExoI. The fourth appears to rely on an allosteric mechanism to disrupt ExoI/SSB complexes. Subsets of the inhibitors block SSB-Ct complex formation with two other SSB-interaction partners as well, which highlights their utility as reagents for investigating the roles of SSB/protein interactions in diverse DNA replication, recombination, and repair reactions. PMID- 20018748 TI - Nanoscale cues regulate the structure and function of macroscopic cardiac tissue constructs. AB - Heart tissue possesses complex structural organization on multiple scales, from macro- to nano-, but nanoscale control of cardiac function has not been extensively analyzed. Inspired by ultrastructural analysis of the native tissue, we constructed a scalable, nanotopographically controlled model of myocardium mimicking the in vivo ventricular organization. Guided by nanoscale mechanical cues provided by the underlying hydrogel, the tissue constructs displayed anisotropic action potential propagation and contractility characteristic of the native tissue. Surprisingly, cell geometry, action potential conduction velocity, and the expression of a cell-cell coupling protein were exquisitely sensitive to differences in the substratum nanoscale features of the surrounding extracellular matrix. We propose that controlling cell-material interactions on the nanoscale can stipulate structure and function on the tissue level and yield novel insights into in vivo tissue physiology, while providing materials for tissue repair. PMID- 20018749 TI - Multiple chromatin-bound protein kinases assemble factors that regulate insulin gene transcription. AB - During the onset of diabetes, pancreatic beta cells become unable to produce sufficient insulin to maintain blood glucose within the normal range. Proinflammatory cytokines have been implicated in impaired beta cell function. To understand more about the molecular events that reduce insulin gene transcription, we examined the effects of hyperglycemia alone and together with the proinflammatory cytokine interleukin-1beta (IL-1beta) on signal transduction pathways that regulate insulin gene transcription. Exposure to IL-1beta in fasting glucose activated multiple protein kinases that associate with the insulin gene promoter and transiently increased insulin gene transcription in beta cells. In contrast, cells exposed to hyperglycemic conditions were sensitized to the inhibitory actions of IL-1beta. Under these conditions, IL 1beta caused the association of the same protein kinases, but a different combination of transcription factors with the insulin gene promoter and began to reduce transcription within 2 h; stimulatory factors were lost, RNA polymerase II was lost, and inhibitory factors were bound to the promoter in a kinase-dependent manner. PMID- 20018750 TI - PPARgamma activation in adipocytes is sufficient for systemic insulin sensitization. AB - Although peroxisome proliferator-activated receptor gamma (PPARgamma) agonists such as thiazolidinediones (TZDs) are widely used to treat type 2 diabetes, how its activation in individual tissues contributes to TZD's therapeutic action remains controversial. As TZDs are known to have receptor-independent effects, we sought to establish gain-of-function animal models to delineate the receptor's insulin-sensitizing actions. Unexpectedly, we find that selective activation of PPARgamma in adipocytes, but not in macrophages, is sufficient for whole-body insulin sensitization equivalent to systemic TZD treatment. In addition to improved adipokine, inflammatory, and lipid profiles, PPARgamma activation in mature adipocytes normalizes serum insulin without increased adipogenesis. Co culture studies indicated that PPARgamma-activated adipocytes broadly suppress induction of inflammatory cytokines and C-X-C family chemokines in macrophages. Collectively, these data describe an "adipocentric" model in which adipose activation of PPARgamma is sufficient for complete insulin sensitization and suggest a specific application for fat selective PPARgamma modulators in diabetic therapy. PMID- 20018751 TI - Mast cells regulate homeostatic intestinal epithelial migration and barrier function by a chymase/Mcpt4-dependent mechanism. AB - Altered intestinal barrier function is postulated to be a central predisposing factor to intestinal diseases, including inflammatory bowel diseases and food allergies. However, the mechanisms involved in maintaining homeostatic intestinal barrier integrity remain undefined. In this study, we demonstrate that mice deficient in mast cells (Kit(W-sh/W-sh) [Wsh]) or mast cell chymase (Mcpt4(-/-)) have significantly decreased basal small intestinal permeability compared with wild-type (WT) mice. Altered intestinal barrier function was linked to decreased intestinal epithelial cell migration along the villus/crypt axis, altered intestinal morphology, and dysregulated claudin-3 crypt expression. Remarkably, engraftment of Wsh mice with WT but not Mcpt4(-/-) mast cells restored intestinal epithelial cell migration, morphology, and intestinal epithelial barrier function. Collectively, these findings identify a mechanism by which mast cells regulate homeostatic intestinal epithelial migration and barrier function. PMID- 20018752 TI - Rotational dynamics of confined C60 from near-infrared Raman studies under high pressure. AB - Peapods present a model system for studying the properties of dimensionally constrained crystal structures, whose dynamical properties are very important. We have recently studied the rotational dynamics of C(60) molecules confined inside single walled carbon nanotube (SWNT) by analyzing the intermediate frequency mode lattice vibrations using near-infrared Raman spectroscopy. The rotation of C(60) was tuned to a known state by applying high pressure, at which condition C(60) first forms dimers at low pressure and then forms a single-chain, nonrotating, polymer structure at high pressure. In the latter state the molecules form chains with a 2-fold symmetry. We propose that the C(60) molecules in SWNT exhibit an unusual type of ratcheted rotation due to the interaction between C(60) and SWNT in the "hexagon orientation," and the characteristic vibrations of ratcheted rotation becomes more obvious with decreasing temperature. PMID- 20018754 TI - Quantitative imaging of the Dorsal nuclear gradient reveals limitations to threshold-dependent patterning in Drosophila. AB - The NF-kappaB-related transcription factor, Dorsal, forms a nuclear concentration gradient in the early Drosophila embryo, patterning the dorsal-ventral (DV) axis to specify mesoderm, neurogenic ectoderm, and dorsal ectoderm cell fates. The concentration of nuclear Dorsal is thought to determine these patterning events; however, the levels of nuclear Dorsal have not been quantified previously. Furthermore, existing models of Dorsal-dependent germ layer specification and patterning consider steady-state levels of Dorsal relative to target gene expression patterns, yet both Dorsal gradient formation and gene expression are dynamic. We devised a quantitative imaging method to measure the Dorsal nuclear gradient while simultaneously examining Dorsal target gene expression along the DV axis. Unlike observations from other insects such as Tribolium, we find the Dorsal gradient maintains a constant bell-shaped distribution during embryogenesis. We also find that some classical Dorsal target genes are located outside the region of graded Dorsal nuclear localization, raising the question of whether these genes are direct Dorsal targets. Additionally, we show that Dorsal levels change in time during embryogenesis such that a steady state is not reached. These results suggest that the multiple gene expression outputs observed along the DV axis do not simply reflect a steady-state Dorsal nuclear gradient. Instead, we propose that the Dorsal gradient supplies positional information throughout nuclear cycles 10-14, providing additional evidence for the idea that compensatory combinatorial interactions between Dorsal and other factors effect differential gene expression along the DV axis. PMID- 20018755 TI - Secretagogin is a Ca2+-binding protein specifying subpopulations of telencephalic neurons. AB - The Ca(2+)-binding proteins (CBPs) parvalbumin, calbindin, and calretinin are phenotypic markers of terminally differentiated neurons in the adult brain. Although subtle phylogenetic variations in the neuronal distribution of these CBPs may occur, morphologically and functionally diverse subclasses of interneurons harbor these proteins in olfactory and corticolimbic areas. Secretagogin (scgn) is a recently cloned CBP from pancreatic beta and neuroendocrine cells. We hypothesized that scgn is expressed in the mammalian brain. We find that scgn is a marker of neuroblasts commuting in the rostral migratory stream. Terminally differentiated neurons in the olfactory bulb retain scgn expression, with scgn being present in periglomerular cells and granular layer interneurons. In the corticolimbic system, scgn identifies granule cells distributed along the dentate gyrus, indusium griseum, and anterior hippocampal continuation emphasizing the shared developmental origins, and cytoarchitectural and functional similarities of these neurons. We also uncover unexpected phylogenetic differences in scgn expression, since this CBP is restricted to primate cholinergic basal forebrain neurons. Overall, we characterize scgn as a neuron-specific CBP whose distribution identifies neuronal subtypes and hierarchical organizing principles in the mammalian brain. PMID- 20018753 TI - Positive selection identifies an in vivo role for FimH during urinary tract infection in addition to mannose binding. AB - FimH, the type 1 pilus adhesin of uropathogenic Escherichia coli (UPEC), contains a receptor-binding domain with an acidic binding pocket specific for mannose. The fim operon, and thus type 1 pilus production, is under transcriptional control via phase variation of an invertible promoter element. FimH is critical during urinary tract infection for mediating colonization and invasion of the bladder epithelium and establishment of intracellular bacterial communities (IBCs). In silico analysis of FimH gene sequences from 279 E. coli strains identified specific amino acids evolving under positive selection outside of its mannose binding pocket. Mutating two of these residues (A27V/V163A) had no effect on phase variation, pilus assembly, or mannose binding in vitro. However, compared to wild-type, this double mutant strain exhibited a 10,000-fold reduction in mouse bladder colonization 24 h after inoculation and was unable to form IBCs even though it bound normally to mannosylated receptors in the urothelium. In contrast, the single A62S mutation altered phase variation, reducing the proportion of piliated cells, reduced mannose binding 8-fold, and decreased bladder colonization 30-fold in vivo compared to wild-type. A phase-locked ON A62S mutant restored virulence to wild-type levels even though in vitro mannose binding remained impaired. Thus, positive selection analysis of FimH has separated mannose binding from in vivo fitness, suggesting that IBC formation is critical for successful infection of the mammalian bladder, providing support for more general use of in silico positive selection analysis to define the molecular underpinnings of bacterial pathogenesis. PMID- 20018756 TI - Complex regulation of the TIR1/AFB family of auxin receptors. AB - Auxin regulates most aspects of plant growth and development. The hormone is perceived by the TIR1/AFB family of F-box proteins acting in concert with the Aux/IAA transcriptional repressors. Arabidopsis plants that lack members of the TIR1/AFB family are auxin resistant and display a variety of growth defects. However, little is known about the functional differences between individual members of the family. Phylogenetic studies reveal that the TIR1/AFB proteins are conserved across land plant lineages and fall into four clades. Three of these subgroups emerged before separation of angiosperms and gymnosperms whereas the last emerged before the monocot-eudicot split. This evolutionary history suggests that the members of each clade have distinct functions. To explore this possibility in Arabidopsis, we have analyzed a range of mutant genotypes, generated promoter swap transgenic lines, and performed in vitro binding assays between individual TIR1/AFB and Aux/IAA proteins. Our results indicate that the TIR1/AFB proteins have distinct biochemical activities and that TIR1 and AFB2 are the dominant auxin receptors in the seedling root. Further, we demonstrate that TIR1, AFB2, and AFB3, but not AFB1 exhibit significant posttranscriptional regulation. The microRNA miR393 is expressed in a pattern complementary to that of the auxin receptors and appears to regulate TIR1/AFB expression. However our data suggest that this regulation is complex. Our results suggest that differences between members of the auxin receptor family may contribute to the complexity of auxin response. PMID- 20018757 TI - Topography and physiology of ascending streams in the auditory tectothalamic pathway. AB - Auditory information is relayed from the cochlea along parallel pathways and reaches the inferior colliculus (IC) and the medial geniculate body (MGB) en route to the cortex. Although the ascending tectothalamic pathway to the ventral division of the MGB is regarded as a high-fidelity information-bearing channel, the roles of the pathways to the dorsal and medial divisions are more opaque. Here, we show fundamental differences between these ascending pathways using an in vitro slice preparation. Using photostimulation, we found three main patterns of input (excitatory, inhibitory, and mixed) that differed in each pathway. Furthermore, electrical stimulation of the central nucleus of the IC evoked a depressing response in the MGB with no metabotropic glutamate (mGlu) receptor component, whereas stimulation of the lateral cortex of the IC evoked a facilitating response with an mGlu receptor component. These data suggest that the ascending tectothalamic pathways are functionally distinct from one another. PMID- 20018758 TI - Inhibition of bacterial disulfide bond formation by the anticoagulant warfarin. AB - Blood coagulation in humans requires the activity of vitamin K epoxide reductase (VKOR), the target of the anticoagulant warfarin (Coumadin). Bacterial homologs of VKOR were recently found to participate in a pathway leading to disulfide bond formation in secreted proteins of many bacteria. Here we show that the VKOR homolog from the bacterium Mycobacterium tuberculosis, the causative agent of human tuberculosis, is inhibited by warfarin and that warfarin-resistant mutations of mycobacterial VKOR appear in similar locations to mutations found in human patients who require higher doses of warfarin. Deletion of VKOR results in a severe growth defect in mycobacteria, and the growth of M. tuberculosis is inhibited by warfarin. The bacterial VKOR homolog may represent a target for antibiotics and a model for genetic studies of human VKOR. We present a simple assay in Escherichia coli, based on a disulfide-sensitive beta-galactosidase, which can be used to screen for stronger inhibitors of the M. tuberculosis VKOR homolog. PMID- 20018759 TI - miR-221 overexpression contributes to liver tumorigenesis. AB - MicroRNA (miRNAs) are negative regulators of gene expression and can function as tumor suppressors or oncogenes. Expression patterns of miRNAs and their role in the pathogenesis of hepatocellular carcinoma (HCC) are still poorly understood. We profiled miRNA expression in tissue samples (104 HCC, 90 adjacent cirrhotic livers, 21 normal livers) as well as in 35 HCC cell lines. A set of 12 miRNAs (including miR-21, miR-221/222, miR-34a, miR-519a, miR-93, miR-96, and let-7c) was linked to disease progression from normal liver through cirrhosis to full blown HCC. miR-221/222, the most up-regulated miRNAs in tumor samples, are shown to target the CDK inhibitor p27 and to enhance cell growth in vitro. Conversely, these activities can be efficiently inhibited by an antagomiR specific for miR 221. In addition, we show, using a mouse model of liver cancer, that miR-221 overexpression stimulates growth of tumorigenic murine hepatic progenitor cells. Finally, we identified DNA damage-inducible transcript 4 (DDIT4), a modulator of mTOR pathway, as a bona fide target of miR-221. Taken together, these data reveal an important contribution for miR-221 in hepatocarcinogenesis and suggest a role for DDIT4 dysregulation in this process. Thus, the use of synthetic inhibitors of miR-221 may prove to be a promising approach to liver cancer treatment. PMID- 20018760 TI - Functional screening identifies CRLF2 in precursor B-cell acute lymphoblastic leukemia. AB - The prognosis for adults with precursor B-cell acute lymphoblastic leukemia (B ALL) remains poor, in part from a lack of therapeutic targets. We identified the type I cytokine receptor subunit CRLF2 in a functional screen for B-ALL-derived mRNA transcripts that can substitute for IL3 signaling. We demonstrate that CRLF2 is overexpressed in approximately 15% of adult and high-risk pediatric B-ALL that lack MLL, TCF3, TEL, and BCR/ABL rearrangements, but not in B-ALL with these rearrangements or other lymphoid malignancies. CRLF2 overexpression can result from translocation with the IGH locus or intrachromosomal deletion and is associated with poor outcome. CRLF2 overexpressing B-ALLs share a transcriptional signature that significantly overlaps with a BCR/ABL signature, and is enriched for genes involved in cytokine receptor and JAK-STAT signaling. In a subset of cases, CRLF2 harbors a Phe232Cys gain-of-function mutation that promotes constitutive dimerization and cytokine independent growth. A mutually exclusive subset harbors activating mutations in JAK2. In fact, all 22 B-ALLs with mutant JAK2 that we analyzed overexpress CRLF2, distinguishing CRLF2 as the key scaffold for mutant JAK2 signaling in B-ALL. Expression of WT CRLF2 with mutant JAK2 also promotes cytokine independent growth that, unlike CRLF2 Phe232Cys or ligand induced signaling by WT CRLF2, is accompanied by JAK2 phosphorylation. Finally, cells dependent on CRLF2 signaling are sensitive to small molecule inhibitors of either JAKs or protein kinase C family kinases. Together, these findings implicate CRLF2 as an important factor in B-ALL with diagnostic, prognostic, and therapeutic implications. PMID- 20018761 TI - Isolation and characterization of centroacinar/terminal ductal progenitor cells in adult mouse pancreas. AB - The question of whether dedicated progenitor cells exist in adult vertebrate pancreas remains controversial. Centroacinar cells and terminal duct (CA/TD) cells lie at the junction between peripheral acinar cells and the adjacent ductal epithelium, and are frequently included among cell types proposed as candidate pancreatic progenitors. However these cells have not previously been isolated in a manner that allows formal assessment of their progenitor capacities. We have found that a subset of adult CA/TD cells are characterized by high levels of ALDH1 enzymatic activity, related to high-level expression of both Aldh1a1 and Aldh1a7. This allows their isolation by FACS using a fluorogenic ALDH1 substrate. FACS-isolated CA/TD cells are relatively depleted of transcripts associated with differentiated pancreatic cell types. In contrast, they are markedly enriched for transcripts encoding Sca1, Sdf1, c-Met, Nestin, and Sox9, markers previously associated with progenitor populations in embryonic pancreas and other tissues. FACS-sorted CA/TD cells are uniquely able to form self-renewing "pancreatospheres" in suspension culture, even when plated at clonal density. These spheres display a capacity for spontaneous endocrine and exocrine differentiation, as well as glucose-responsive insulin secretion. In addition, when injected into cultured embryonic dorsal pancreatic buds, these adult cells display a unique capacity to contribute to both the embryonic endocrine and exocrine lineages. Finally, these cells demonstrate dramatic expansion in the setting of chronic epithelial injury. These findings suggest that CA/TD cells are indeed capable of progenitor function and may contribute to the maintenance of tissue homeostasis in adult mouse pancreas. PMID- 20018762 TI - NCLX is an essential component of mitochondrial Na+/Ca2+ exchange. AB - Mitochondrial Ca(2+) efflux is linked to numerous cellular activities and pathophysiological processes. Although it is established that an Na(+)-dependent mechanism mediates mitochondrial Ca(2+) efflux, the molecular identity of this transporter has remained elusive. Here we show that the Na(+)/Ca(2+) exchanger NCLX is enriched in mitochondria, where it is localized to the cristae. Employing Ca(2+) and Na(+) fluorescent imaging, we demonstrate that mitochondrial Na(+) dependent Ca(2+) efflux is enhanced upon overexpression of NCLX, is reduced by silencing of NCLX expression by siRNA, and is fully rescued by the concomitant expression of heterologous NCLX. NCLX-mediated mitochondrial Ca(2+) transport was inhibited, moreover, by CGP-37157 and exhibited Li(+) dependence, both hallmarks of mitochondrial Na(+)-dependent Ca(2+) efflux. Finally, NCLX-mediated mitochondrial Ca(2+) exchange is blocked in cells expressing a catalytically inactive NCLX mutant. Taken together, our results converge to the conclusion that NCLX is the long-sought mitochondrial Na(+)/Ca(2+) exchanger. PMID- 20018763 TI - FoxP3+ regulatory T cells essentially contribute to peripheral CD8+ T-cell tolerance induced by steady-state dendritic cells. AB - Peripheral T-cell tolerance is thought to significantly contribute to the prevention of autoimmunity, and it has been shown that antigen-presenting steady state dendritic cells efficiently induce peripheral tolerance. We previously showed that dendritic-cell-induced tolerance is a T-cell-intrinsic process that depends on coinhibitory molecules such as programmed death-1. Here we specifically analyze the involvement of FoxP3(+) regulatory T cells, which are known to be important for maintenance of self-tolerance. We show that antigen presentation by steady-state dendritic cells failed to induce peripheral tolerance in the absence of FoxP3(+) regulatory T cells but induced protective CD8(+) T-cell-mediated immunity instead. Regulatory T-cell-depleted mice had massively increased numbers of dendritic cells in lymph nodes. Dendritic cells isolated from mice without regulatory T cells had up-regulated costimulatory molecules and showed stronger T-cell stimulatory capacity ex vivo, suggesting that regulatory T cells contribute to peripheral tolerance by keeping the dendritic cells in an immature state. Using blocking antibodies, we demonstrate that CTLA-4 but not IL-10 is necessary for control of dendritic cells by regulatory T cells. PMID- 20018764 TI - Stochastic fate of p53-mutant epidermal progenitor cells is tilted toward proliferation by UV B during preneoplasia. AB - UV B (UVB) radiation induces clones of cells mutant for the p53 tumor suppressor gene in human and murine epidermis. Here we reanalyze large datasets that report the fate of clones in mice subjected to a course of UVB radiation, to uncover how p53 mutation affects epidermal progenitor cell behavior. We show that p53 mutation leads to exponential growth of clones in UV-irradiated epidermis; this finding is also consistent with the size distribution of p53 mutant clones in human epidermis. Analysis of the tail of the size distribution further reveals that the fate of individual mutant cells is stochastic. Finally, the data suggest that ending UVB exposure results in the p53 mutant cells adopting the balanced fate of wild-type cells: the loss of mutant cells is balanced by proliferation so that the population of preneoplastic cells remains constant. We conclude that preneoplastic clones do not derive from long-lived, self-renewing mutant stem cells but rather from mutant progenitors with random cell fate. It follows that ongoing, low-intensity UVB radiation will increase the number of precancerous cells dramatically compared with sporadic, higher-intensity exposure at the same cumulative dose, which may explain why nonmelanoma skin cancer incidence depends more strongly on age than on radiation dosage. Our approach may be applied to determine cell growth rates in clonally labeled material from a wide range of tissues including human samples. PMID- 20018765 TI - Quantifying cellular traction forces in three dimensions. AB - Cells engage in mechanical force exchange with their extracellular environment through tension generated by the cytoskeleton. A method combining laser scanning confocal microscopy (LSCM) and digital volume correlation (DVC) enables tracking and quantification of cell-mediated deformation of the extracellular matrix in all three spatial dimensions. Time-lapse confocal imaging of migrating 3T3 fibroblasts on fibronectin (FN)-modified polyacrylamide gels of varying thickness reveals significant in-plane (x, y) and normal (z) displacements, and illustrates the extent to which cells, even in nominally two-dimensional (2-D) environments, explore their surroundings in all three dimensions. The magnitudes of the measured displacements are independent of the elastic moduli of the gels. Analysis of the normal displacement profiles suggests that normal forces play important roles even in 2-D cell migration. PMID- 20018766 TI - Systems analysis of plant cell wall degradation by the model filamentous fungus Neurospora crassa. AB - The filamentous fungus Neurospora crassa is a model laboratory organism, but in nature is commonly found growing on dead plant material, particularly grasses. Using functional genomics resources available for N. crassa, which include a near full genome deletion strain set and whole genome microarrays, we undertook a system-wide analysis of plant cell wall and cellulose degradation. We identified approximately 770 genes that showed expression differences when N. crassa was cultured on ground Miscanthus stems as a sole carbon source. An overlap set of 114 genes was identified from expression analysis of N. crassa grown on pure cellulose. Functional annotation of up-regulated genes showed enrichment for proteins predicted to be involved in plant cell wall degradation, but also many genes encoding proteins of unknown function. As a complement to expression data, the secretome associated with N. crassa growth on Miscanthus and cellulose was determined using a shotgun proteomics approach. Over 50 proteins were identified, including 10 of the 23 predicted N. crassa cellulases. Strains containing deletions in genes encoding 16 proteins detected in both the microarray and mass spectrometry experiments were analyzed for phenotypic changes during growth on crystalline cellulose and for cellulase activity. While growth of some of the deletion strains on cellulose was severely diminished, other deletion strains produced higher levels of extracellular proteins that showed increased cellulase activity. These results show that the powerful tools available in N. crassa allow for a comprehensive system level understanding of plant cell wall degradation mechanisms used by a ubiquitous filamentous fungus. PMID- 20018767 TI - The SAH domain extends the functional length of the myosin lever. AB - Stable, single alpha-helix (SAH) domains are widely distributed in the proteome, including in myosins, but their functions are unknown. To test whether SAH domains can act as levers, we replaced four of the six calmodulin-binding IQ motifs in the levers of mouse myosin 5a (Myo5) with the putative SAH domain of Dictyostelium myosin MyoM of similar length. The SAH domain was inserted between the IQ motifs and the coiled coil in a Myo5 HMM construct in which the levers were truncated from six to two IQ motifs (Myo5-2IQ). Electron microscopy of this chimera (Myo5-2IQ-SAH) showed the SAH domain was straight and 17 nm long as predicted, restoring the truncated lever to the length of wild-type (Myo5-6IQ). The powerstroke (of 21.5 nm) measured in the optical trap was slightly less than that for Myo5-6IQ but much greater than for Myo5-2IQ. Myo5-2IQ-SAH moved processively along actin at physiological ATP concentrations with similar stride and run lengths to Myo5-6IQ in in-vitro single molecule assays. In comparison, Myo5-2IQ is not processive under these conditions. Solution biochemical experiments indicated that the rear head did not mechanically gate the rate of ADP release from the lead head, unlike Myo5-6IQ. These data show that the SAH domain can form part of a functional lever in myosins, although its mechanical stiffness might be lower. More generally, we conclude that SAH domains can act as stiff structural extensions in aqueous solution and this structural role may be important in other proteins. PMID- 20018768 TI - Modulation of Fgf3 dosage in mouse and men mirrors evolution of mammalian dentition. AB - A central challenge in evolutionary biology is understanding how genetic mutations underlie morphological changes. Because highly calcified enamel enables preservation of detailed dental features, studying tooth morphology enables this question to be addressed in both extinct and extant species. Previous studies have found that mutant mice can have severe abnormalities in tooth morphology, and several authors have explored the evolutionary implications of tooth number modifications in mutants. However, although they can potentially shed much light on evolutionary mechanisms, anomalies in tooth shape remain poorly studied. Here, we report that alterations in dosage of the Fgf3 gene cause morphological changes in both genetically engineered mutant mice and in human patients. By comparing the dental morphologies in mice and humans carrying Fgf3 mutations with primitive rodent and primate fossils, we determined that decreases in dosage of Fgf3 lead to phenotypes that resemble the progressive reappearance of ancestral morphologies. We propose that modifications in the FGF signaling pathway have played an important role in evolution of mammalian dentition by giving rise to new cusps and interconnecting cusps by new crests. We anticipate that our multidisciplinary study will advance the detailed correlation of subtle dental modifications with genetic mutations in a variety of mammalian lineages. PMID- 20018769 TI - The importance of being quantitative. PMID- 20018770 TI - Adaptive response by state-dependent inactivation. AB - Many membrane channels and receptors exhibit adaptive, or desensitized, response to a strong sustained input stimulus. A key mechanism that underlies this response is the slow, activity-dependent removal of responding molecules to a pool which is unavailable to respond immediately to the input. This mechanism is implemented in different ways in various biological systems and has traditionally been studied separately for each. Here we highlight the common aspects of this principle, shared by many biological systems, and suggest a unifying theoretical framework. We study theoretically a class of models which describes the general mechanism and allows us to distinguish its universal from system-specific features. We show that under general conditions, regardless of the details of kinetics, molecule availability encodes an averaging over past activity and feeds back multiplicatively on the system output. The kinetics of recovery from unavailability determines the effective memory kernel inside the feedback branch, giving rise to a variety of system-specific forms of adaptive response-precise or input-dependent, exponential or power-law-as special cases of the same model. PMID- 20018771 TI - Can molecular biology and bioinformatics be used to probe an evolutionary pathway? PMID- 20018772 TI - Cross-correlations between volume change and price change. AB - In finance, one usually deals not with prices but with growth rates R, defined as the difference in logarithm between two consecutive prices. Here we consider not the trading volume, but rather the volume growth rate R, the difference in logarithm between two consecutive values of trading volume. To this end, we use several methods to analyze the properties of volume changes |R|, and their relationship to price changes |R|. We analyze 14,981 daily recordings of the Standard and Poor's (S & P) 500 Index over the 59-year period 1950-2009, and find power-law cross-correlations between |R| and |R| by using detrended cross correlation analysis (DCCA). We introduce a joint stochastic process that models these cross-correlations. Motivated by the relationship between |R| and |R|, we estimate the tail exponent alpha of the probability density function P(|R|) approximately |R|(-1-alpha) for both the S & P 500 Index as well as the collection of 1819 constituents of the New York Stock Exchange Composite Index on 17 July 2009. As a new method to estimate alpha, we calculate the time intervals tau(q) between events where R > q. We demonstrate that tau(q), the average of tau(q), obeys tau(q) approximately q(alpha). We find alpha approximately 3. Furthermore, by aggregating all tau(q) values of 28 global financial indices, we also observe an approximate inverse cubic law. PMID- 20018773 TI - Optical single-channel resolution imaging of the ryanodine receptor distribution in rat cardiac myocytes. AB - We have applied an optical super-resolution technique based on single-molecule localization to examine the peripheral distribution of a cardiac signaling protein, the ryanodine receptor (RyR), in rat ventricular myocytes. RyRs form clusters with a mean size of approximately 14 RyRs per cluster, which is almost an order of magnitude smaller than previously estimated. Clusters were typically not circular (as previously assumed) but elongated with an average aspect ratio of 1.9. Edge-to-edge distances between adjacent RyR clusters were often <50 nm, suggesting that peripheral RyR clusters may exhibit strong intercluster signaling. The wide variation of cluster size, which follows a near-exponential distribution, is compatible with a stochastic cluster assembly process. We suggest that calcium sparks may be the result of the concerted activation of several RyR clusters forming a functional "supercluster" whose gating is controlled by both cytosolic and sarcoplasmic reticulum luminal calcium levels. PMID- 20018774 TI - Food webs are more than the sum of their tritrophic parts. AB - Many studies have aimed to understand food webs by investigating components such as trophic links (one consumer taxon eats one resource taxon), tritrophic interactions (one consumer eats an intermediate taxon, which eats a resource), or longer chains of links. We show here that none of these components (links, tritrophic interactions, and longer chains), individually or as an ensemble, accounts fully for the properties of the next higher level of organization. As a cell is more than its molecules, as an organ is more than its cells, and as an organism is more than its organs, in a food web, new structure emerges at every organizational level up to and including the whole web. We demonstrate the emergence of properties at progressively higher levels of structure by using all of the directly observed, appropriately organized, publicly available food web datasets with relatively complete trophic link data and with average body mass and population density data for each taxon. There are only three such webs, those of Tuesday Lake, Michigan, in 1984 and 1986, and Ythan Estuary, Scotland. We make the data freely available online with this report. Differences in web patterns between Tuesday Lake and Ythan Estuary, and similarities of Tuesday Lake in 1984 and 1986 despite 50% turnover of species, suggest that the patterns we describe respond to major differences between ecosystem types. PMID- 20018775 TI - Host structural carbohydrate induces vector transmission of a bacterial plant pathogen. AB - Many insect-borne pathogens have complex life histories because they must colonize both hosts and vectors for successful dissemination. In addition, the transition from host to vector environments may require changes in gene expression before the pathogen's departure from the host. Xylella fastidiosa is a xylem-limited plant-pathogenic bacterium transmitted by leafhopper vectors that causes diseases in a number of economically important plants. We hypothesized that factors of host origin, such as plant structural polysaccharides, are important in regulating X. fastidiosa gene expression and mediating vector transmission of this pathogen. The addition of pectin and glucan to a simple defined medium resulted in dramatic changes in X. fastidiosa's phenotype and gene expression profile. Cells grown in the presence of pectin became more adhesive than in other media tested. In addition, the presence of pectin and glucan in media resulted in significant changes in the expression of several genes previously identified as important for X. fastidiosa's pathogenicity in plants. Furthermore, vector transmission of X. fastidiosa was induced in the presence of both polysaccharides. Our data show that host structural polysaccharides mediate gene regulation in X. fastidiosa, which results in phenotypic changes required for vector transmission. A better understanding of how vector-borne pathogens transition from host to vector, and vice versa, may lead to previously undiscovered disease-control strategies. PMID- 20018776 TI - The impact of soil microorganisms on the global budget of delta18O in atmospheric CO2. AB - Improved global estimates of terrestrial photosynthesis and respiration are critical for predicting the rate of change in atmospheric CO(2). The oxygen isotopic composition of atmospheric CO(2) can be used to estimate these fluxes because oxygen isotopic exchange between CO(2) and water creates distinct isotopic flux signatures. The enzyme carbonic anhydrase (CA) is known to accelerate this exchange in leaves, but the possibility of CA activity in soils is commonly neglected. Here, we report widespread accelerated soil CO(2) hydration. Exchange was 10-300 times faster than the uncatalyzed rate, consistent with typical population sizes for CA-containing soil microorganisms. Including accelerated soil hydration in global model simulations modifies contributions from soil and foliage to the global CO(18)O budget and eliminates persistent discrepancies existing between model and atmospheric observations. This enhanced soil hydration also increases the differences between the isotopic signatures of photosynthesis and respiration, particularly in the tropics, increasing the precision of CO(2) gross fluxes obtained by using the delta(18)O of atmospheric CO(2) by 50%. PMID- 20018777 TI - The D-type cyclin CYCD4;1 modulates lateral root density in Arabidopsis by affecting the basal meristem region. AB - Root cell division occurs primarily in the apical meristem, from which cells are displaced into the basal meristem, where division decreases and cell length increases before the final differentiation zone. The organization of the root in concentric files implies coordinated division and differentiation of cell types, including the xylem pole pericycle cells, which uniquely can resume division to initiate lateral roots (LR). Here, we show that D-type cyclin CYCD4;1 is expressed in meristematic pericycle protoxylem poles and is required for normal LR density. Cycd4;1 mutants also show a displacement of the apical/basal meristem boundary in the pericycle and longer pericycle basal meristem cells, whereas other cell layers and overall meristem size and root growth are unaffected. Auxin is proposed to separately prepattern and stimulate LR initiation. Stimulation is unimpaired in cycd4;1, suggesting CYCD4;1 requirement for normal spacing but not initiation. Both pericycle cell length and LR density phenotypes of cycd4;1 are rescued by low concentrations of applied auxin, suggesting that the basal meristem has a role in determining LR density. We further show CYCD4;1 is rate limiting for sucrose-dependent LR formation, since CYCD4;1 expression is sucrose dependent and wild-type roots fully phenocopy cycd4;1 in sucrose absence. We conclude that CYCD4;1 links meristem pericycle cell behavior to LR density consistent with a basal meristem prepatterning model and that D-type cyclins can confer division potential of defined cell types through cell-specific expression patterns. PMID- 20018778 TI - Distributions of observed death tolls govern sensitivity to human fatalities. AB - How we react to humanitarian crises, epidemics, and other tragic events involving the loss of human lives depends largely on the extent to which we are moved by the size of their associated death tolls. Many studies have demonstrated that people generally exhibit a diminishing sensitivity to the number of human fatalities and, equivalently, a preference for risky (vs. sure) alternatives in decisions under risk involving human losses. However, the reason for this tendency remains unknown. Here we show that the distributions of event-related death tolls that people observe govern their evaluations of, and risk preferences concerning, human fatalities. In particular, we show that our diminishing sensitivity to human fatalities follows from the fact that these death tolls are approximately power-law distributed. We further show that, by manipulating the distribution of mortality-related events that people observe, we can alter their risk preferences in decisions involving fatalities. Finally, we show that the tendency to be risk-seeking in mortality-related decisions is lower in countries in which high-mortality events are more frequently observed. Our results support a model of magnitude evaluation based on memory sampling and relative judgment. This model departs from the utility-based approaches typically encountered in psychology and economics in that it does not rely on stable, underlying value representations to explain valuation and choice, or on choice behavior to derive value functions. Instead, preferences concerning human fatalities emerge spontaneously from the distributions of sampled events and the relative nature of the evaluation process. PMID- 20018779 TI - Embryonic stem cell tumor model reveals role of vascular endothelial receptor tyrosine phosphatase in regulating Tie2 pathway in tumor angiogenesis. AB - Inhibiting angiogenesis has become an effective approach for treating cancer and other diseases. However, our understanding of signaling pathways in tumor angiogenesis has been limited by the embryonic lethality of many gene knockouts. To overcome this limitation, we used the plasticity of embryonic stem (ES) cells to develop a unique approach to study tumor angiogenesis. Murine ES cells can be readily manipulated genetically; in addition, ES cells implanted subcutaneously in mice develop into tumors that contain a variety of cell types (teratomas). We show that ES cells differentiate into bona fide endothelial cells within the teratoma, and that these ES-derived endothelial cells form part of the functional tumor vasculature. Using this powerful and flexible system, the Angiopoietin/Tie2 system is shown to have a key role in the regulation of tumor vessel size. Endothelial differentiation in the ES teratoma model allows gene-targeting methods to be used in the study of tumor angiogenesis. PMID- 20018780 TI - Projections of future sea level becoming more dire. PMID- 20018781 TI - New perspectives of information transformation through the auditory cortical layers. PMID- 20018782 TI - What is bet-hedging, really? PMID- 20018783 TI - Inheritance of nesting behaviour across natural environmental variation in a turtle with temperature-dependent sex determination. AB - Nesting behaviour is critical for reproductive success in oviparous organisms with no parental care. In organisms where sex is determined by incubation temperature, nesting behaviour may be a prime target of selection in response to unbalanced sex ratios. To produce an evolutionary change in response to sex-ratio selection, components of nesting behaviour must be heritable. We estimated the field heritability of two key components of nesting behaviour in a population of painted turtles (Chrysemys picta) with temperature-dependent sex determination by applying the 'animal model' to a pedigree reconstructed from genotype data. We obtained estimates of low to non-detectable heritability using repeated records across all environments. We then determined environment-specific heritability by grouping records with similar temperatures for the winter preceding the nesting season, a variable known to be highly associated with our two traits of interest, nest vegetation cover and Julian date of nesting. The heritability estimates of nest vegetation cover and Julian date of nesting were qualitatively highest and significant, or nearly so, after hot winters. Additive genetic variance for these traits was not detectable after cold winters. Our analysis suggests that the potential for evolutionary change of nesting behaviour may be dependent on the thermal conditions of the preceding winter, a season that is predicted to be especially subject to climate change. PMID- 20018784 TI - Avian population consequences of climate change are most severe for long-distance migrants in seasonal habitats. AB - One consequence of climate change is an increasing mismatch between timing of food requirements and food availability. Such a mismatch is primarily expected in avian long-distance migrants because of their complex annual cycle, and in habitats with a seasonal food peak. Here we show that insectivorous long-distance migrant species in The Netherlands declined strongly (1984-2004) in forests, a habitat characterized by a short spring food peak, but that they did not decline in less seasonal marshes. Also, within generalist long-distance migrant species, populations declined more strongly in forests than in marshes. Forest-inhabiting migrant species arriving latest in spring declined most sharply, probably because their mismatch with the peak in food supply is greatest. Residents and short distance migrants had non-declining populations in both habitats, suggesting that habitat quality did not deteriorate. Habitat-related differences in trends were most probably caused by climate change because at a European scale, long-distance migrants in forests declined more severely in western Europe, where springs have become considerably warmer, when compared with northern Europe, where temperatures during spring arrival and breeding have increased less. Our results suggest that trophic mismatches may have become a major cause for population declines in long-distance migrants in highly seasonal habitats. PMID- 20018785 TI - Multiple signals and male spacing affect female preference at cocktail parties in treefrogs. AB - Effective acoustic communication in the face of intense conspecific background noise constitutes a constant sensory challenge in chorusing and colonial species. An evolutionary approach suggests that behavioural and environmental constraints in these species should have shaped signal design and signalling behaviour to enable communication in noisy conditions. This could be attained both through the use of multicomponent signals and through short-term adjustments in the spatial separation of calling males. We investigated these two hypotheses in a chorusing anuran, the hylid Hyla arborea, through a series of phonotaxis experiments conducted within a six-speaker arena in a high background noise situation, by presenting females with male calls containing either single or multiple attractive call components, and by modifying distances between speakers. We found that female ability to discriminate attractive calls increased when several attractive call components were available, providing novel evidence that the use of multicomponent signals enhances communication in complex acoustic conditions. Signal discrimination in females also improved with speaker separation, demonstrating that within natural choruses, spatial unmasking conditioned by male density and spatial separation probably improves female discrimination of competing males. Implications of these results for the accuracy of mate choice within choruses are discussed. PMID- 20018786 TI - Transverse stresses and modes of failure in tree branches and other beams. AB - The longitudinal stresses in beams subjected to bending also set up transverse stresses within them; they compress the cross section when the beam's curvature is being increased and stretch it when its curvature is being reduced. Analysis shows that transverse stresses rise to a maximum at the neutral axis and increase with both the bending moment applied and the curvature of the beam. These stresses can qualitatively explain the fracture behaviour of tree branches. Curved 'hazard beams' that are being straightened split down the middle because of the low transverse tensile strength of wood. By contrast, straight branches of light wood buckle when they are bent because of its low transverse compressive strength. Branches of denser wood break, but the low transverse tensile strength diverts the crack longitudinally when the fracture has only run half-way across the beam, to produce their characteristic 'greenstick fracture'. The bones of young mammals and uniaxially reinforced composite beams may also be prone to greenstick fracture because of their lower transverse tensile strength. PMID- 20018787 TI - Optic flow informs distance but not profitability for honeybees. AB - How do flying insects monitor foraging efficiency? Honeybees (Apis mellifera) use optic flow information as an odometer to estimate distance travelled, but here we tested whether optic flow informs estimation of foraging costs also. Bees were trained to feeders in flight tunnels such that bees experienced the greatest optic flow en route to the feeder closest to the hive. Analyses of dance communication showed that, as expected, bees indicated the close feeder as being further, but they also indicated this feeder as the more profitable, and preferentially visited this feeder when given a choice. We show that honeybee estimates of foraging cost are not reliant on optic flow information. Rather, bees can assess distance and profitability independently and signal these aspects as separate elements of their dances. The optic flow signal is sensitive to the nature of the environment travelled by the bee, and is therefore not a good index of flight energetic costs, but it provides a good indication of distance travelled for purpose of navigation and communication, as long as the dancer and recruit travel similar routes. This study suggests an adaptive dual processing system in honeybees for communicating and navigating distance flown and for evaluating its energetic costs. PMID- 20018788 TI - Selective biodegradation of keratin matrix in feather rachis reveals classic bioengineering. AB - Flight necessitates that the feather rachis is extremely tough and light. Yet, the crucial filamentous hierarchy of the rachis is unknown-study hindered by the tight chemical bonding between the filaments and matrix. We used novel microbial biodegradation to delineate the fibres of the rachidial cortex in situ. It revealed the thickest keratin filaments known to date (factor >10), approximately 6 microm thick, extending predominantly axially but with a small outer circumferential component. Near-periodic thickened nodes of the fibres are staggered with those in adjacent fibres in two- and three-dimensional planes, creating a fibre-matrix texture with high attributes for crack stopping and resistance to transverse cutting. Close association of the fibre layer with the underlying 'spongy' medulloid pith indicates the potential for higher buckling loads and greater elastic recoil. Strikingly, the fibres are similar in dimensions and form to the free filaments of the feather vane and plumulaceous and embryonic down, the syncitial barbules, but, identified for the first time in 140+ years of study in a new location-as a major structural component of the rachis. Early in feather evolution, syncitial barbules were consolidated in a robust central rachis, definitively characterizing the avian lineage of keratin. PMID- 20018789 TI - Diversity trends and their ontogenetic basis: an exploration of allometric disparity in rodents. AB - It has been hypothesized that most morphological evolution occurs by allometric differentiation. Because rodents encapsulate a phenomenal amount of taxonomic diversity and, among several clades, contrasting levels of morphological diversity, they represent an excellent subject to address the question: how variable are allometric patterns during evolution? We investigated the influence of phylogenetic relations and ecological factors on the results of the first quantification of allometric disparity among rodents by exploring allometric space, a multivariate morphospace here derived from, and encapsulating all, the ontogenetic trajectories of 34 rodent species from two parallel phylogenetic radiations. Disparity was quantified using angles between ontogenetic trajectories for different species and clades. We found an overlapping occupation of allometric space by muroid and hystricognath species, revealing both clades possess similar abilities to evolve in different directions of phenotypic space, and anatomical diversity does not act to constrain the labile nature of allometric patterning. Morphological features to enable efficient processing of food serve to group rodents in allometric space, reflecting the importance of convergent morphology, rather than shared evolutionary history, in the generation of allometric patterns. Our results indicate that the conserved level of morphological integration found among primates cannot simply be extended to all mammals. PMID- 20018790 TI - A photoactivatable green-fluorescent protein from the phylum Ctenophora. AB - Genes for the family of green-fluorescent proteins (GFPs) have been found in more than 100 species of animals, with some species containing six or more copies producing a variety of colours. Thus far, however, these species have all been within three phyla: Cnidaria, Arthropoda and Chordata. We have discovered GFP type fluorescent proteins in the phylum Ctenophora, the comb jellies. The ctenophore proteins share the xYG chromophore motif of all other characterized GFP-type proteins. These proteins exhibit the uncommon property of reversible photoactivation, in which fluorescent emission becomes brighter upon exposure to light, then gradually decays to a non-fluorescent state. In addition to providing potentially useful optical probes with novel properties, finding a fluorescent protein in one of the earliest diverging metazoans adds further support to the possibility that these genes are likely to occur throughout animals. PMID- 20018791 TI - Fitts' law is not continuous in reciprocal aiming. AB - It takes longer to accomplish difficult tasks than easy ones. In the context of motor behaviour, Fitts' famous law states that the time needed to successfully execute an aiming movement increases linearly with task difficulty. While Fitts' explicit formulation has met criticism, the relation between task difficulty and movement time is invariantly portrayed as continuous. Here, we demonstrate that Fitts' law is discontinuous in reciprocal aiming owing to a transition in operative motor control mechanisms with increasing task difficulty. In particular, rhythmic movements are implemented in easy tasks and discrete movements in difficult ones. How movement time increases with task difficulty differs in both movement types. It appears, therefore, that the human nervous system abruptly engages a different control mechanism when task difficulty increases. PMID- 20018792 TI - Bet-hedging as an evolutionary game: the trade-off between egg size and number. PMID- 20018793 TI - Implications of flexible-shelled eggs in a Cretaceous choristoderan reptile. AB - Flexible, or soft-shelled, eggs are almost unknown in the fossil record, leaving large gaps in our knowledge of the reproductive biology of many tetrapod clades. Here, we report two flexible-shelled eggs of the hyphalosaurid choristodere Hyphalosaurus baitaigouensis from the Early Cretaceous of China, one containing an embryo and the second associated with a neonate. Choristoderes are an enigmatic group of aquatic reptiles that survived the K-T extinction but died out in the Miocene. Hyphalosaurids, a specialized clade of Choristodera, resemble miniature plesiosaurs and are considered to be primarily aquatic in habit. Scanning electron microscopy of samples from the eggs reveals a thin, non columnar external mineralized layer characterized by rounded nodes and tentatively identified poorly structured irregular pores, with an underlying amorphous layer presumably representing decomposed protein fibrils. While the relationships of Choristodera remain controversial, eggshell microstructure more closely resembles that of Lepidosauromorpha (the lineage including lizards) as opposed to that of Archosauromorpha (the lineage including birds and crocodiles). PMID- 20018794 TI - Productive activities and development of frailty in older adults. AB - OBJECTIVE: Our aim was to examine whether engagement in productive activities, including volunteering, paid work, and childcare, protects older adults against the development of geriatric frailty. METHODS: Data from the first (1988) and second (1991) waves of the MacArthur Study of Successful Aging, a prospective cohort study of high-functioning older adults aged 70-79 years (n = 1,072), was used to examine the hypothesis that engagement in productive activities is associated with lower levels of frailty 3 years later. RESULTS: Engagement in productive activities at baseline was associated with a lower cumulative odds of frailty 3 years later in unadjusted models (odds ratio [OR] = 0.74, 95% confidence interval [CI] = 0.58-0.96) but not after adjusting for age, disability, and cognitive function (adjusted OR = 0.78, 95% CI = 0.60-1.01). Examination of productive activity domains showed that volunteering (but neither paid work nor childcare) was associated with a lower cumulative odds of frailty after adjusting for age, disability, and cognitive function. This relationship diminished and was no longer statistically significant after adjusting for personal mastery and religious service attendance. DISCUSSION: Though high functioning older adults who participate in productive activities are less likely to become frail, after adjusting for age, disability, and cognitive function, only volunteering is associated with a lower cumulative odds of frailty. PMID- 20018795 TI - Overall and anterior Bolton ratio in Class I, II, and III orthodontic patients. AB - The aim of the investigation was to compare overall and anterior Bolton ratios in different malocclusion groups with Bolton's standards. The material comprised 600 pre-treatment study casts (262 males and 338 females, aged 12-25 years), selected from the models of 3088 patients who had applied for orthodontic treatment based on the following criteria: permanent dentition from the first right molar to the first left molar and no interproximal caries or restorations. There were 162 Class I, 144 Class II division 1, 155 Class II division 2, and 139 Class III patients. Statistical analysis of the data was undertaken using a Student's t test. Statistically significant differences were found for the mean overall ratio when compared with the original Bolton norm for the whole study group, as well as for patients with Class I and III malocclusions when the mean anterior ratio was compared with the original Bolton norm. Significant differences were observed in all malocclusion groups for both genders. Discrepancies exceeding 2 SD were found in 31.2 per cent of the studied population for the anterior ratio when compared with Bolton's norm. The highest mean values for anterior ratio were in males with Class I (79.1) and Class III (80.1) malocclusions. PMID- 20018797 TI - Prostaglandin E2 levels in gingival crevicular fluid during tooth- and bone-borne expansion. AB - The purpose of this study was to compare Prostaglandin E(2) (PGE(2)) levels in gingival crevicular fluid (GCF) of young adults with maxillary constriction during tooth- and bone-borne expansion. Thirty patients, 15 females and 15 males, with a mean age of 17.3 +/- 2.8 years were divided into three groups. Group I consisted of 10 patients, five females and five males, treated by transpalatal distraction (TPD) as a bone-borne device, group II 10 patients, five females and five males, with a Hyrax appliance as a tooth-borne device, and a control group of 10 patients, five females and five males, without any expansion appliances. GCF samples were collected with filter paper strips at six observation periods in order to evaluate the effect of heavy orthopaedic forces in both groups. In group II, the samples were additionally collected at two pre-treatment time points in order to evaluate the effect of the forces generated by the separators. An automated enzyme immunoassay was used to measure PGE(2) in the GCF. The differences within the groups were evaluated with a pairwise t-test and the differences between the groups were determined by the Mann-Whitney U-test. The mean PGE(2) level was significantly elevated on day 4 after placement of the separators in group II (P < 0.05). The PGE(2) values in group II were significantly different to those in group I and the controls at all observation periods. Lower PGE(2) levels were observed in group I compared with group II and the controls. Expansion using the TPD method could potentially enhance the prognosis of the teeth by inducing more skeletal dental changes when compared with the Hyrax appliance. PMID- 20018796 TI - Comparison of dentoalveolar protrusion values in Moroccans and other populations. AB - SUMMARY: The aim of this retrospective study was to evaluate bimaxillary protrusion among Moroccans and to compare the Moroccan population with other Arabic, Caucasian, and Mediterranean populations. Cephalometric radiographs of 102 Moroccan adult university students (73 females and 29 males, mean age: 21 years 6 months +/- 1 year 6 months) with a normal occlusion were traced. Fifteen measurements concerning the upper and lower incisors, upper and lower lips, and the Frankfort to mandibular plane angle were assessed. Statistical analysis of the data was undertaken using independent t-tests. Eight angular and six linear measurements were statistically significantly different compared with Caucasian cephalometric norms (P < 0.001). Significant differences were observed compared with other adult Arabic populations (Palestinians and Saudis) and with Turkish Anatolian adults. Relative to Caucasian cephalometric norms, Moroccans showed bimaxillary protrusion and, therefore, these norms cannot be applied to Moroccan faces. In general, there was a great similarity in incisor position between the Moroccan and Saudi populations. PMID- 20018798 TI - A comparison of pain experienced by patients treated with labial and lingual orthodontic appliances. AB - SUMMARY: The aim of this prospective longitudinal study was to compare pain experiences among Chinese adult patients treated with labial and lingual orthodontic appliances. Sixty patients, 30 with labial appliances (18 females and 12 males, mean age 20.33 years, SD +/- 4.205) and 30 with lingual appliances (22 females and 8 males, mean age 21.63 years, SD +/- 2.236), rated their overall pain experience on a 100 mm visual analogue scale (VAS) at three time points: 1 week (T(1)), 1 month (T(2)), and 3 months (T(3)) after bracket placement. In addition, on a separate 100 mm VAS, they rated their pain experience at the locations of the tongue, lips, cheeks, gums, face, and jaw at T(1), T(2), and T(3). Changes in pain VAS were conducted using Friedman analysis of variance, area under the curve (AUC) analysis and the data were compared using a t-test. There was no significant difference in global ratings of pain among those treated with labial or lingual appliances (P > 0.05). Among both groups, global ratings of pain decreased over the study period (P < 0.001). Patients treated with lingual appliances reported higher ratings of tongue pain (P < 0.001), while those treated with labial appliances reported higher ratings of lip (P < 0.001) and cheek (P < 0.001) pain. The findings indicate that patients treated with labial and lingual appliances rate similarly the level of overall pain they experience during treatment. Ratings of overall pain experienced decreased for both treatment groups with time. However, ratings of pain differed at various sites with respect to the type of orthodontic appliance. These findings have implications in informing patients' treatment decision-making processes regarding labial and lingual appliances and in the management of discomfort associated with different treatment modalities. PMID- 20018803 TI - Toward understanding tight glycemic control in the ICU: a systematic review and metaanalysis. AB - BACKGROUND: Following publication of the Leuven Intensive Insulin Therapy Trial in 2001, tight glycemic control became the standard of care in ICUs around the world. Recent studies suggest that this approach may be flawed. The goal of this systematic review was to determine the benefits and risks of tight glycemic control in ICU patients and to explain the differences in outcomes among reported trials. METHODS: Prospective, randomized controlled clinical trials (RCTs) that studied the impact of tight glycemic control (blood glucose 80-110 mg/dL) on mortality in ICU patients were identified through a search of MEDLINE, EMBASE, and the Cochrane Database of Systematic Reviews, and a citation review of relevant primary and review articles. Data were abstracted on study design, study size, and patient characteristics, as well as on the mean (or median) and SD of the ICU blood glucose level, mean daily dose of insulin administered, average daily caloric intake, percentage of calories given intravenously (parenteral nutrition), incidence of hypoglycemia, need for dialysis, and 28-day/hospital mortality. Metaanalytic techniques were used to analyze the data; subgroup analysis and metaregression were used to explain differences in the treatment effect. RESULTS: We identified seven RCT studies that included 11,425 patients. Overall, tight glycemic control did not reduce the 28-day mortality (odds ratio [OR] 0.95; 95% CI, 0.87-1.05), the incidence of blood stream infections (OR 1.04; 95% CI, 0.93-1.17), or the requirement for renal replacement therapy (OR 1.01; 95% CI, 0.89-1.13). The incidence of hypoglycemia was significantly higher in patients randomized to tight glycemic control (OR 7.7; 95% CI, 6.0-9.9; P < .001). Metaregression demonstrated a significant relationship between the treatment effect (28-day mortality) and the proportion of calories provided parenterally (P = .005). This suggests that the difference in outcome between the two Leuven Intensive Insulin Therapy Trials and the subsequent trials could be related to the use of parenteral nutrition. When the two Leuven Intensive Insulin Therapy Trials were excluded from the metaanalysis, mortality was lower in the control patients (OR 0.90; 95% CI, 0.81-0.99; P = .04; I(2) = 0%). CONCLUSIONS: There is no evidence to support the use of intensive insulin therapy in general medical-surgical ICU patients who are fed according to current guidelines. Tight glycemic control is associated with a high incidence of hypoglycemia and an increased risk of death in patients not receiving parenteral nutrition. PMID- 20018804 TI - The natural 13C abundance of plasma glucose is a useful biomarker of recent dietary caloric sweetener intake. AB - There is a need for objective biomarkers of dietary intake, because self reporting is often subject to bias. We tested the validity of a biomarker for the fraction of dietary carbohydrate (CHO) from cane sugar and high fructose corn syrup (C(4) sugars) using natural (13)C abundance of plasma glucose. In a randomized, single-blinded, crossover design, 5 participants consumed 3 weight maintaining diets for 7 d, with a 2-wk washout between diet periods. Diets differed in the fraction of total CHO energy from C(4) sugars (5, 16, or 32%). During each diet period, blood samples were drawn at hours 0800 and 1600 on d 1, 3, and 5 and at 0800, 1000, 1200, 1400, and 1600 on d 7. The delta(13)C abundance of plasma glucose was analyzed via GC- isotope ratio MS. Within each diet period, delta(13)C abundance of the 0800 fasting glucose did not change from baseline with increasing time during a diet period; however, there was a strong positive correlation (R(2) = 0.89) between delta(13)C abundance of the glucose concentration at 1000 on d 7 and the percent of breakfast CHO from C(4) sugars. Also, delta(13)C abundance of the combined plasma glucose samples on d 7 demonstrated a strong positive correlation (R(2) = 0.90) with the percent of total daily CHO from C(4) sugars. The natural delta(13)C abundance of postprandial plasma glucose relative to dietary C(4) CHO content was a valid biomarker for contributions of C(4) caloric sweeteners from the previous meal. PMID- 20018805 TI - Hepatic acyl-coenzyme a:cholesterol acyltransferase-2 expression is decreased in mice with hyperhomocysteinemia. AB - Alterations in lipid metabolism may contribute to the pathology of hyperhomocysteinemia (HHcy). Our objective in this study was to test the hypothesis that HHcy is associated with changes in liver acyl CoA:cholesterol acyl transferase 2 (ACAT2) expression and cholesteryl esters (CE) in mice with HHcy. ACAT2 is encoded by Soat2 and functions to catalyze the esterification of cholesterol with acyl-CoA. Mice heterozygous for disruption of the cystathionine beta-synthase gene (Cbs +/-) and C57BL/6 mice (Cbs +/+) were fed a control diet or a diet high in l-methionine (8.60 g/kg) and low in folic acid (0.20 mg/kg) to induce HHcy (HH diet). Lower Soat2 mRNA (P < 0.05) and ACAT protein (P < 0.001), higher total oleic acid [18:1(n-9)], and lower CE 18:1(n-9) was found in liver from Cbs +/- mice fed the HH diet, with higher plasma total homocysteine concentrations, than Cbs +/+ mice fed the control diet (35.01 +/- 5.6 vs. 2.21 +/ 0.6 mumol/L, respectively). In silico searches identified a CpG-rich region in the 5' portion of the Soat2 gene, which was differentially methylated (P < 0.05) in Cbs +/- mice fed the HH diet than in Cbs +/+ mice fed the control diet and was accompanied by higher (P < 0.05) B1 repeat element methylation, an indicator of global de novo methylation. These findings show altered methylation and expression of Soat2/ACAT2 in liver from mice with HHcy and suggest a role for changes in liver CE in the pathology of HHcy. PMID- 20018806 TI - Probiotic-derived factors: probiotaceuticals? PMID- 20018807 TI - Hibiscus sabdariffa L. tea (tisane) lowers blood pressure in prehypertensive and mildly hypertensive adults. AB - In vitro studies show Hibiscus sabdariffa L., an ingredient found in many herbal tea blends and other beverages, has antioxidant properties, and, in animal models, extracts of its calyces have demonstrated hypocholesterolemic and antihypertensive properties. Our objective in this study was to examine the antihypertensive effects of H. sabdariffa tisane (hibiscus tea) consumption in humans. A randomized, double-blind, placebo-controlled clinical trial was conducted in 65 pre- and mildly hypertensive adults, age 30-70 y, not taking blood pressure (BP)-lowering medications, with either 3 240-mL servings/d of brewed hibiscus tea or placebo beverage for 6 wk. A standardized method was used to measure BP at baseline and weekly intervals. At 6 wk, hibiscus tea lowered systolic BP (SBP) compared with placebo (-7.2 +/- 11.4 vs. -1.3 +/- 10.0 mm Hg; P = 0.030). Diastolic BP was also lower, although this change did not differ from placebo (-3.1 +/- 7.0 vs. -0.5 +/- 7.5 mm Hg; P = 0.160). The change in mean arterial pressure was of borderline significance compared with placebo (-4.5 +/- 7.7 vs. -0.8 +/- 7.4 mm Hg; P = 0.054). Participants with higher SBP at baseline showed a greater response to hibiscus treatment (r = -0.421 for SBP change; P = 0.010). No effects were observed with regard to age, gender, or dietary supplement use. These results suggest daily consumption of hibiscus tea, in an amount readily incorporated into the diet, lowers BP in pre- and mildly hypertensive adults and may prove an effective component of the dietary changes recommended for people with these conditions. PMID- 20018808 TI - Age and dietary iron affect expression of genes involved in iron acquisition and homeostasis in young pigs. AB - To investigate the effects of dietary iron (Fe) and age on Fe metabolism, we used 36 weaned barrows in a 2 x 3 design with 2 concentrations of dietary Fe [97 (control) and 797 (high Fe) mg Fe/kg dry matter] and 3 time points of tissue collection (after 21, 42, or 63 d on diets). Pigs were weighed and bled on d 0, 20, 41, and 62. High Fe reduced feed efficiency but did not affect pig weight gain. Blood hemoglobin concentrations and Fe concentrations of liver, intestine, and heart were increased by high dietary Fe on all days. Concentrations of liver and heart Fe increased with age. As determined by quantitative real-time PCR, hepatic expression of hepcidin (HAMP) in pigs given the high-Fe diet was 6.25 fold that of control pigs. In the intestine, relative mRNA levels of ferroportin, divalent metal transporter 1, and transferrin receptor were downregulated by high Fe. Expression of an alternative route of Fe absorption, solute carrier family 39 member 14 (SLC39A14), was downregulated in the intestine of pigs fed high dietary Fe. Additionally, duodenal mRNA level of certain genes including scavenger receptor class A, member 5, and frataxin decreased with age of the animal. Our findings indicate new roles in Fe metabolism for several mineral metabolism associated genes and that some of these genes, such as SLC39A14, may be regulated in response to dietary Fe in pigs. Additionally, the expression of some genes examined in this study was affected by age, suggesting age dependency of Fe metabolism in pigs. PMID- 20018809 TI - Sildenafil citrate treatment enhances amino acid availability in the conceptus and fetal growth in an ovine model of intrauterine growth restriction. AB - Adequate placental blood flow is essential for the optimal delivery of nutrients from mother to fetus for conceptus growth. Restricted fetal development results from pathophysiological and environmental factors that alter utero-placental blood flow, placental function, and, therefore, nutrient availability in the fetus. To test this hypothesis, 0, 75, or 150 mg/d sildenafil citrate (Viagra) was administered subcutaneously from d 28 to 115 of gestation to either nutrient restricted [50% of NRC requirements) or adequately-fed ewes (100% of NRC requirements). On d 115, maternal, fetal, and placental tissues and fluids were collected. Concentrations of total amino acids and polyamines in uterine venous and arterial sera, amniotic and allantoic fluids, and fetal umbilical venous serum were lower (P < 0.05) in nutrient-restricted ewes than in adequately fed ewes, as were the ratios of total amino acids in fetal umbilical venous serum to uterine arterial serum. Sildenafil citrate dose-dependently increased (P < 0.05) total amino acids and polyamines in amniotic fluid, allantoic fluid, and fetal serum without affecting values in maternal serum. Fetal weight was lower (P < 0.05) in nutrient-restricted ewes on d 115. Sildenafil citrate treatment dose dependently increased (P < 0.05) fetal weight in both nutrient-restricted and adequately fed ewes. This study supports the hypothesis that long-term sildenafil citrate treatment enhances fetal growth, at least in part, by increasing the availability of amino acids in the conceptus. These findings may lead to the clinical use of sildenafil citrate in human pregnancies suspected to be at risk for intrauterine fetal growth retardation. PMID- 20018810 TI - Phosphatidylinositol 3,4,5-triphosphate-dependent Rac exchanger 1 (P-Rex-1), a guanine nucleotide exchange factor for Rac, mediates angiogenic responses to stromal cell-derived factor-1/chemokine stromal cell derived factor-1 (SDF-1/CXCL 12) linked to Rac activation, endothelial cell migration, and in vitro angiogenesis. AB - Stromal cell-derived factor-1 (SDF-1/CXCL-12) and vascular endothelial growth factor (VEGF), which can be secreted by hypoxic tumors, promote the generation of new blood vessels. These potent angiogenic factors stimulate endothelial cell migration via the activation of Rho GTPases and the phosphatidylinositol 3-kinase (PI3K)/AKT signaling pathway. Thus, characterization of guanine nucleotide exchange factors critical in the angiogenic signaling cascades offers the possibility of identifying novel molecular targets. We demonstrated previously that mammalian target of rapamycin, an important effector and regulator of PI3K/AKT, activates phosphatidylinositol 3,4,5-triphosphate-dependent Rac exchanger 1 (P-Rex1), a Rac guanine nucleotide exchange factor identified as a target of G betagamma and PI3K, via direct interactions. In this study, we tested the hypothesis that P-Rex1 is involved in the angiogenic responses elicited by SDF-1 and VEGF. Using a knockdown approach, we demonstrate that P-Rex1 is indeed required for SDF-1 promoted signaling pathway, because there is decreased Rac activation, cell migration, and in vitro angiogenesis in P-Rex1 knockdown cells stimulated with SDF-1. In contrast, P-Rex1 knockdown does not affect responses to VEGF, and signaling to extracellular signal-regulated kinase in response to either angiogenic factor is not sensitive to P-Rex1 knockdown. We also demonstrate that in endothelial cells, VEGF promotes an increase in the expression of endogenous P-Rex1 and the SDF-1 receptor CXCR4, In addition, VEGF pretreated cells show an increased migratory and angiogenic response to SDF-1, suggesting that VEGF stimulation can complement SDF-1/CXCR4 signaling to induce angiogenesis. We conclude that P-Rex1 is a key element in SDF-1-induced angiogenic responses and a potential target for therapeutic intervention. PMID- 20018811 TI - The small GTPase Ral couples the angiotensin II type 1 receptor to the activation of phospholipase C-delta 1. AB - The angiotensin II type 1 receptor (AT(1)R) plays an important role in cardiovascular function and as such represents a primary target for therapeutic intervention. The AT(1)R has traditionally been considered to be coupled to the activation of phospholipase C (PLC) beta via its association with G alpha(q/11), leading to increases in intracellular inositol phosphate (IP) and release of calcium from intracellular stores. In the present study, we investigated whether the small GTPase RalA contributed to the regulation of AT(1)R endocytosis and signaling. We find that neither RalA nor RalB is required for the endocytosis of the AT(1)R, but that RalA expression is required for AT(1)R-stimulated IP formation but not 5-HT(2A) receptor-mediated IP formation. AT(1)R-activated IP formation is lost in the absence of Ral guanine nucleotide dissociation stimulator (RalGDS), and requires the beta-arrestin-dependent plasma membrane translocation of RalGDS. G alpha(q/11) small interfering RNA (siRNA) treatment also significantly attenuates both AT(1)R- and 5-HT(2A) receptor-stimulated IP formation after 30 min of agonist stimulation. PLC-delta1 has been reported to be activated by RalA, and we show that AT(1)R-stimulated IP formation is attenuated after PLC-delta 1 siRNA treatment. Taken together, our results provide evidence for a G protein-coupled recepto-activated and RalGDS/Ral-mediated mechanism for PLC-delta 1 stimulation. PMID- 20018812 TI - Tirucallic acids are novel pleckstrin homology domain-dependent Akt inhibitors inducing apoptosis in prostate cancer cells. AB - Activation of the serine/threonine kinase Akt is associated with aggressive clinical behavior of prostate cancer. We found that the human prostate cancer cell lines LNCaP and PC-3 express predominantly Akt1 and Akt2. Selective down regulation of Akt1, but not Akt2, by short-hairpin RNA reduced the viability of prostate cancer cells. In addition, structurally different Akt inhibitors were cytotoxic for the prostate cancer cells, confirming that the Akt pathway is indispensable for their viability. We have purified the tetracyclic triterpenoids 3-oxo-tirucallic acid, 3-alpha-acetoxy-tirucallic acid, and 3-beta-acetoxy tirucallic acid from the oleogum resin of Boswellia carterii to chemical homogeneity. The acetoxy-derivatives in particular potently inhibited the activities of human recombinant Akt1 and Akt2 and of constitutively active Akt immunoprecipitated from PC-3 cells, whereas inhibitor of nuclear factor-kappaB kinases remained unaffected. Docking data indicated that these tetracyclic triterpenoids form hydrogen bonds within the phosphatidylinositol binding pocket of the Akt pleckstrin homology domain. Accordingly, 3-beta-acetoxy-tirucallic acid did not inhibit the activity of Akt1 lacking the pleckstrin homology domain. In the prostate cancer cell lines investigated, these compounds inhibited the phosphorylation of cellular Akt and the Akt signaling pathways, including glycogen synthase kinase-3beta and BAD phosphorylation, nuclear accumulation of p65, the androgen receptor, beta-catenin, and c-Myc. These events culminated in the induction of apoptosis in prostate cancer, but not in nontumorigenic cells. The tirucallic acid derivatives inhibited proliferation and induced apoptosis in tumors xenografted onto chick chorioallantoic membranes and decreased the growth of pre-established prostate tumors in nude mice without overt systemic toxicity. Thus, tirucallic acid derivatives represent a new class of Akt inhibitors with antitumor properties. PMID- 20018813 TI - Carbapenem susceptibility patterns for clinical isolates of Mycobacterium abscessus determined by the Etest method. AB - Mycobacterium abscessus is resistant to multiple antibiotics, creating treatment challenges. Carbapenems are tested to increase therapeutic alternatives. We performed in vitro susceptibility testing by Etest of four carbapenems for M. abscessus isolates. Imipenem demonstrated the most in vitro activity, and testing of other carbapenems provided no additional value. PMID- 20018814 TI - Mutability in the matrix gene of novel influenza A H1N1 virus detected using a FRET probe-based real-time reverse transcriptase PCR assay. PMID- 20018815 TI - Molecular epidemiology of pediatric pneumococcal empyema from 2001 to 2007 in Utah. AB - Utah had a high rate of pediatric pneumococcal empyema (PPE) prior to licensure of the pneumococcal conjugate vaccine (PCV-7) in 2000. The majority (62%) of PPE cases was due to nonvaccine serotypes, primarily Streptococcus pneumoniae serotype 1, multilocus sequence type (MLST) 227. PPE in Utah children has increased over the last decade. It is unclear whether the increase was due to serotype replacement or switch. In this study, we describe the incidence and molecular epidemiology of PPE by MLST in Utah children after the licensure of PCV 7. Empyema rates increased from 8.5/100,000 children in the state of Utah in 2001 to 12.5/100,000 children in 2007 (P = 0.006). Ninety-eight percent was due to nonvaccine serotypes (P < 0.001 when compared to the pre-PCV-7 period). PPE was primarily due to serotypes 1, 3, 19A, and 7F, with MLST demonstrating sequence types (ST) that were commonly present in the United States prior to licensure of PCV-7. Serotype switch was not documented. Replacement disease with common ST of serotypes 1,3, 7F, and 19A rather than serotype switch was responsible for the increase in PPE in Utah children. PMID- 20018816 TI - Genetic diversity of Mycobacterium tuberculosis isolates from a tertiary care tuberculosis hospital in South Korea. AB - Tuberculosis (TB) remains an immense public health problem in the Republic of Korea despite a more than fivefold decrease in the prevalence of the disease over the last 3 decades. The rise in drug-resistant TB has compounded the situation. We analyzed 208 clinical isolates of M. tuberculosis from the National Masan Tuberculosis Hospital by spoligotyping, IS6110 restriction fragment length polymorphism (RFLP), and 24-locus-based mycobacterial interspersed repetitive unit-variable number tandem repeat (MIRU-VNTR) typing to assess the diversity and transmission dynamics of the tubercle bacilli in the Republic of Korea. The majority of the isolates (97.1%) belonged to the Beijing genotype. Cluster analysis by MIRU-VNTR yielded a low clustering rate of 22.3%, with most of the clusters comprising isolates with diverse drug resistance patterns. The discriminatory capacity of the typing methods was high for RFLP and MIRU-VNTR (allelic diversity [h] = 0.99) but low for spoligotyping (h = 0.31). Although analysis of 19 MIRU-VNTR loci was needed to achieve maximum discrimination, an informative set of 8 loci (960, 1955, 2163b, 2165, 2996, 3192, 4052, and 4348) (h = 0.98) that was able to differentiate most of the closely related strains was identified. These findings suggest that 24-locus-based MIRU-VNTR typing is a likely suitable alternative to RFLP to differentiate clinical isolates in this setting, which is dominated by M. tuberculosis Beijing strains. Within the study limits, our results also suggest that the problem of drug-resistant TB in the Republic of Korea may be largely due to acquired resistance as opposed to transmission. PMID- 20018818 TI - Nonhemolytic Streptococcus pyogenes isolates that lack large regions of the sag operon mediating streptolysin S production. AB - Among nonhemolytic Streptococcus pyogenes (group A streptococcus) strains (n = 9) isolated from patients with pharyngitis or acute otitis media, we identified three deletions in the region from the epf gene, encoding the extracellular matrix binding protein, to the sag operon, mediating streptolysin S production. PMID- 20018817 TI - Use of fimH single-nucleotide polymorphisms for strain typing of clinical isolates of Escherichia coli for epidemiologic investigation. AB - Strain typing methods that compare electrophoresis banding patterns are commonly used but are difficult to standardize and poorly portable. Multilocus sequence typing (MLST) is a sequence-based alternative, but it is not practical for large scale epidemiological studies. In the present study, the usefulness of fimH single-nucleotide polymorphisms (SNPs) for Escherichia coli typing was explored. fimH SNPs were determined for 345 E. coli clinical isolates (including 3 reference strains) and compared to PCR-based ECOR (E. coli reference collection) phylogrouping. The fimH gene could be amplified for 316 (92%) of the 345 isolates. fimH SNP analysis found 46 distinct terminal groups in the nucleotide sequence-based phylogenetic tree (fimH types). A subset of the E. coli isolates (162 clinical isolates and the 3 reference strains) were compared by fimH type, PCR phylogroup, and MLST. These isolates fell into 27 fimH types and 18 MLST clonal complexes (CCs) that contained 2 to 28 isolates per complex. The combination of PCR phylogroup and fimH type corresponded to a single CC for 113 (68%) isolates and 2 or 3 CCs for the other 52 (32%) isolates. We propose that the combination of PCR phylogrouping and fimH SNP analysis may be a useful method to type a large collection of clinical E. coli isolates for epidemiologic studies. PMID- 20018819 TI - Molecular evidence of persistent epidemic and evolution of subgenotype B1 coxsackievirus A16-associated hand, foot, and mouth disease in China. AB - The molecular epidemiology of CVA16 in China between 1999 and 2008 reflects a pattern of endemic cocirculation of clusters B1a and B1b within subgenotype B1 viruses. The annual evolution rate of CVA16 was estimated as approximately 0.91 x 10(-2) substitutions per synonymous nucleotide/year and is slightly lower than that of HEV71. PMID- 20018820 TI - Asymmetric dimethylarginine in angiotensin II-induced hypertension. AB - Recent studies have shown that asymmetric dimethylarginine (ADMA), a nitric oxide synthase inhibitor, is increased in hypertension and chronic kidney disease. However, little is known about the effects of hypertension per se on ADMA metabolism. The purpose of this study was to test the hypothesis that ANG II induced hypertension, in the absence of renal injury, is associated with increased oxidative stress and plasma and renal cortex ADMA levels in rats. Male Sprague-Dawley rats were treated with ANG II at 200 ng.kg(-1).min(-1) sc (by minipump) for 1 or 3 wk or at 400 ng.kg(-1).min(-1) for 6 wk. Mean arterial pressure was increased after 3 and 6 wk of ANG II; however, renal injury (proteinuria, glomerular sclerosis, and interstitial fibrosis) was only evident after 6 wk of treatment. Plasma thiobarbituric acid reactive substances concentration and renal cortex p22(phox) protein abundance were increased early (1 and 3 wk), but urinary excretion of isoprostane and H(2)O(2) was only increased after 6 wk of ANG II. An increased in plasma ADMA after 6 wk of ANG II was associated with increased lung protein arginine methyltransferase-1 abundance and decreased renal cortex dimethylarginine dimethylaminohydrolase activity. No changes in renal cortex ADMA were observed. ANG II hypertension in the absence of renal injury is not associated with increased ADMA; however, when the severity and duration of the treatment were increased, plasma ADMA increased. These data suggest that elevated blood pressure alone, for up to 3 wk, in the absence of renal injury does not play an important role in the regulation of ADMA. However, the presence of renal injury and sustained hypertension for 6 wk increases ADMA levels and contributes to nitric oxide deficiency and cardiovascular disease. PMID- 20018821 TI - The 24-hour respiratory quotient predicts energy intake and changes in body mass. AB - To define the relationship between the respiratory quotient (RQ) and energy intake (EI) and to determine the impact of spontaneous locomotor activity (LMA) in the development of diet-induced obesity (DIO), we fed C57BL/6 mice a high-fat diet (HFD) for either 4 days or 17 wk and analyzed them using indirect calorimetry. Importantly, changes in body mass during calorimetry (DeltaM(b)) significantly covaried with RQ and EI; adjusting the data for DeltaM(b) permitted an analysis of the energy-balanced state. The 24-h RQ strongly predicted 24-h EI, and the slope of this relationship was diet dependent (HFD or chow) but independent of the HFD feeding period. Early-stage DIO was characterized by dark period hyperphagia and fat storage, offset by greater light-period lipid oxidation; later stage DIO mice had a milder hyperphagia and lower substrate flexibility. Consequently, whereas 24-h RQ equaled the food quotient of the HFD in both early- and late-stage DIO, the range of RQ values was negatively correlated with, and mostly explained by, 24-h EI only in late-stage DIO. Lean and early-stage DIO mice had similar LMA values that were reduced in late-stage DIO. However, LMA significantly explained variance in total energy expenditure (EE) in only early-stage DIO mice. This indicated that the link between LMA and EE was a transient adaptive response to early DIO, whereas the later loss of LMA did not explain body weight gain in C57BL/6 DIO mice. PMID- 20018822 TI - Refractory blood pressure in female SHR to increased oxidative stress is not mediated by NO or by upregulation of renal antioxidant enzymes. AB - There is a sex difference in the blood pressure (BP) responses to prooxidants and antioxidants in the spontaneously hypertensive rat (SHR). In contrast to males, BP in female SHR does not decrease in response to antioxidants, such as tempol or apocynin, or increase in response to the prooxidant, molsidomine. Molsidomine decreases BP and increases expression of antioxidants in male Wistar-Kyoto rats (WKY), but not male SHR. The present study tested the hypothesis that the mechanism responsible for the lack of a pressor response to molsidomine in females is due to higher endogenous nitric oxide (NO) or to compensatory upregulation of renal antioxidant enzymes. Female SHR were treated with molsidomine in the presence or absence of nitro-L-arginine methyl ester (L-NAME) for 2 wk. Molsidomine increased nitrate/nitrite (NO(x)) and F2-isoprostane (F2 IsoP) excretion, whereas L-NAME reduced NO(x) but increased F-Isop. Molsidomine and L-NAME together further reduced NO(x) and increased F2-IsoP. Molsidomine alone had no effect on BP; L-NAME alone increased BP. The combination of molsidomine and L-NAME did not increase BP above L-NAME alone levels. Whole body and renal oxidative stress increased, while renal cortical Cu,Zn-SOD expression was downregulated and catalase was upregulated by molsidomine; glutathione peroxidase expression was unaffected. These data support our previous studies suggesting that BP in female SHR is independent of either increases or decreases in oxidative stress. The mechanisms responsible for the sex difference in BP response to increase or decrease of oxidative stress are not due to increased NO in females or to compensatory upregulation of antioxidant enzymes in response to increases in oxidants. PMID- 20018823 TI - Aging impairs the expression of the catalytic subunit of glutamate cysteine ligase in soleus muscle under stress. AB - This study investigated the mechanisms responsible for the disrupted homeostasis of reduced glutathione (GSH) in aging muscles with stress (14 days of hind-limb unloading [HU]). Adult and old rats were randomized into four groups: weight bearing and 3, 7, and 14 days of HU. Soleus muscles were harvested to investigate the activity or content of enzymes involved in GSH metabolism (utilization and synthesis). The activities of glutathione S transferase, glutathione reductase, gamma-glutamyl transpeptidase, and glutamate cysteine ligase (GCL) were determined. The protein content of the two subunits of GCL, catalytic subunit (GCLC) and modifier subunit (GCLM), were evaluated. The major results, failure to maintain the accelerated GCLC production and GCL activity, are associated with the GSH depletion in aging muscles with 14 days of HU. The results suggest that the regulation of GCL, especially the catalytic subunit, with stress may be compromised in aging muscles. PMID- 20018824 TI - Comorbidities, functional limitations, and geriatric syndromes in relation to treatment and survival patterns among elders with colorectal cancer. AB - PURPOSE: To examine patterns of colorectal cancer (CRC) treatment and survival in relation to comorbidities (COM), functional limitations (FL), and geriatric syndromes (GS). METHODS: Our study population consisted of Ohio elders diagnosed with incident invasive CRC in the period August 1999 to November 2001 and admitted to home health care (HHC) in the 30 days before or after cancer diagnosis (n = 1009). We used data from the Ohio Cancer Incidence Surveillance System, vital records, and Medicare administrative data, including the HHC Outcome and Assessment Information Set (OASIS), which includes detailed clinical data for HHC patients. Counts of COM, FL, and GS at baseline were retrieved from the OASIS. Multivariable logistic and survival models were developed to examine the association between clinical attributes and outcomes, adjusting for demographic covariates and cancer stage. RESULTS: Comorbidities were associated with increased likelihood of surgery-only, but not with surgery + chemotherapy. Both FL and GS were associated with lower likelihood to undergo surgery-only or surgery + chemotherapy. Two or more GS was associated with disease-specific mortality (adjusted hazard ratio [AHR]: 2.71; 95% confidence interval [CI]: 1.80 4.07) and overall mortality (AHR: 2.34; 95% CI: 1.74-3.15). Two or more FL was associated with overall mortality (AHR: 1.33; 95% CI: 1.10-1.62), but not with disease-specific mortality. COM was not associated with overall mortality, but was negatively associated with disease-specific mortality at borderline level of significance (AHR: 0.78; 95% CI: 0.61-1.00). CONCLUSION: Our findings demonstrate the importance of accounting for FL and GS, in addition to COM, when studying cancer-related outcomes in elders. PMID- 20018825 TI - Plasma cytokine levels in a population-based study: relation to age and ethnicity. AB - BACKGROUND: Aging is one factor believed to contribute to processes that underlie chronic low-grade inflammation in older adults. Moreover, more recent studies have suggested that cytokine levels are influenced by ethnicity. METHODS: In this study, we determined plasma cytokine profiles in a population-based sample (n = 1,411; aged 25-91 years) to determine the relationship between circulating cytokine levels, aging, and ethnicity. We measured interleukin-1 receptor antagonist (IL-1ra), interleukin (IL)-6, -10, C-reactive protein (CRP), and tumor necrosis factor-receptor 1 (TNF-r1). RESULTS: IL-6 and TNF-r1 significantly increased with age, whereas IL-1ra, IL-10, and CRP did not significantly increase with age. After adjusting for age, non-Hispanic whites had significantly higher levels of IL-1ra than Mexican Americans, whereas non-Hispanic blacks had significantly higher levels of IL-6 and CRP than Mexican Americans as well as non Hispanic whites. CRP levels in non-Hispanic blacks were no longer significantly higher after adjusting for body mass index (BMI), indicating that BMI is an important predictor of this inflammatory marker. CONCLUSIONS: These results demonstrate that cytokine levels are influenced by both age and ethnicity. Furthermore, these results show that inflammatory profiles for Mexican Americans are lower than non-Hispanic whites and non-Hispanic blacks. PMID- 20018826 TI - Low serum free triiodothyronine levels mark familial longevity: the Leiden Longevity Study. AB - BACKGROUND: The hypothalamo-pituitary-thyroid axis has been widely implicated in modulating the aging process. Life extension effects associated with low thyroid hormone levels have been reported in multiple animal models. In human populations, an association was observed between low thyroid function and longevity at old age, but the beneficial effects of low thyroid hormone metabolism at middle age remain elusive. METHODS: We have compared serum thyroid hormone function parameters in a group of middle-aged offspring of long-living nonagenarian siblings and a control group of their partners, all participants of the Leiden Longevity Study. RESULTS: When compared with their partners, the group of offspring of nonagenarian siblings showed a trend toward higher serum thyrotropin levels (1.65 vs157 mU/L, p = .11) in conjunction with lower free thyroxine levels (15.0 vs 15.2 pmol/L, p = .045) and lower free triiodothyronine levels (4.08 vs 4.14 pmol/L, p = .024). CONCLUSIONS: Compared with their partners, the group of offspring of nonagenarian siblings show a lower thyroidal sensitivity to thyrotropin. These findings suggest that the favorable role of low thyroid hormone metabolism on health and longevity in model organism is applicable to humans as well. PMID- 20018827 TI - Differences in functional impairment across subtypes of dementia. AB - BACKGROUND: Dementia is a cause of disability in later life. Despite the importance of functional status to the diagnosis of dementia, limited information exists on differences in functional limitations by dementia subtype. We conducted a cross-sectional analysis using the Aging, Demographics, and Memory Study (ADAMS) to determine the extent of functional impairment among older adults with dementia due to different etiologies. METHODS: The ADAMS sample consisted of 856 individuals aged 71 years and older originally surveyed as part of the Health and Retirement Study. Based on a comprehensive in-person cognitive evaluation, respondents were assigned to diagnostic categories of normal cognition, cognitive impairment not demented, and demented. Dementia subtypes were grouped into three categories: vascular dementia (VaD), Alzheimer's dementia (AD), and dementia due to other etiologies. For 744 of the 856 respondents, a proxy informant completed a questionnaire asking whether the respondent had difficulty completing instrumental activities of daily living and activities of daily living (ADLs). RESULTS: Of 744 ADAMS participants, 263 had dementia: 199 (70.5%) with AD, 42 (16.9%) with VaD, and 22 (12.6%) were demented due to other etiologies. After adjustment for demographics, chronic illnesses, and dementia severity, participants with VaD (odds ratio [OR] 5.74; 95% confidence interval [CI] 2.60 12.69) and other etiologies of dementia (OR 21.23; 95% CI 7.25-62.16) were more likely to have greater than or equal to four ADL limitations compared with those with AD. CONCLUSIONS: VaD is associated with significantly more ADL limitations than AD. These physical limitations should be considered when designing adult day care programs, which adequately accommodate the needs of non-AD patients. PMID- 20018828 TI - A contribution of area 5 of the posterior parietal cortex to the planning of visually guided locomotion: limb-specific and limb-independent effects. AB - We tested the hypothesis that area 5 of the posterior parietal cortex (PPC) contributes to the planning of visually guided gait modifications. We recorded 121 neurons from the PPC of two cats during a task in which cats needed to process visual input to step over obstacles attached to a moving treadmill belt. During unobstructed locomotion, 64/121 (53%) of cells showed rhythmic activity. During steps over the obstacles, 102/121 (84%) of cells showed a significant change of their activity. Of these, 46/102 were unmodulated during the control task. We divided the 102 task-related cells into two groups on the basis of their discharge when the limb contralateral to the recording site was the first to pass over the obstacle. One group (41/102) was characterized by a brief, phasic discharge as the lead forelimb passed over the obstacle (Step-related cells). These cells were recorded primarily from area 5a. The other group (61/102) showed a progressive increase in activity prior to the onset of the swing phase in the modified limb and frequently diverged from control at least one step cycle before the gait modification (Step-advanced cells). Most of these cells were recorded in area 5b. In both groups, some cells maintained a fixed relationship to the activity of the contralateral forelimb regardless of which limb was the first to pass over the obstacle (limb-specific cells), whereas others changed their phase of activity so that they were always related to activity of the first limb to pass over the obstacle, either contralateral or ipsilateral (limb-independent cells). Limb-independent cells were more common among the Step-advanced cell population. We suggest that both populations of cells contribute to the gait modification and that the discharge characteristics of the Step-advanced cells are compatible with a contribution to the planning of the gait modification. PMID- 20018829 TI - Varying overall sound intensity to the two ears impacts interaural level difference discrimination thresholds by single neurons in the lateral superior olive. AB - The lateral superior olive (LSO) is one of the earliest sites in the auditory pathway involved in processing acoustical cues to sound location. LSO neurons encode the interaural level difference (ILD) cue to azimuthal location. Here we investigated the effect of variations in the overall stimulus levels of sounds at the two ears on the sensitivity of LSO neurons to small differences in ILDs of pure tones. The neuronal firing rate versus ILD functions were found to depend greatly on the overall stimulus level, typically shifting along the ILD axis toward the excitatory ear and attaining greater maximal firing rates as stimulus level increased. Seventy-five percent of neurons showed significant shifts with changes in overall sound level. The range of ILDs corresponding to best neural acuity for ILDs shifted accordingly. In a simulation using the empirical data, when the overall stimulus level was randomly changed from one trial to the next, the neural discrimination thresholds for ILD, or ILD acuities, were worsened by 50-60% across the population of neurons relative to fixed stimulus levels whether ILD acuity was measured at the azimuthal midline or the ILD pedestal producing the best acuity. The impairment in ILD discrimination was attributed to the increased neural response variance imparted by varying the stimulus level. These results contrast to those observed in psychophysical studies where ILD discrimination thresholds under similar experimental conditions are invariant to overall changes in stimulus level. A simple computational model that incorporated the antagonistic inputs of bilateral LSO nuclei as well as the dorsal nuclei of the lateral lemniscus to the inferior colliculus produced a more robust encoding of ILD even in the setting of roving stimulus level. Testable predictions of this model and comparison to other computational models addressing stimulus invariance were considered. PMID- 20018830 TI - Role of neurotransmitter receptors in mediating light-evoked responses in retinal interplexiform cells. AB - Interplexiform (IP) cells are a long-neglected group of retinal neurons the function of which is yet to be determined. Anatomical study indicates that IP cells are located in the inner nuclear layer, juxtaposed with the third-order neurons. However, the synaptic transmission of IP cells in the inner retina is poorly understood. Using whole cell patch-clamp and pharmacological techniques, we extensively studied synaptic receptors in IP cells. The IP cells in amphibian retinal slices were identified by electrical and morphological properties with voltage-clamp recording and Lucifer yellow dialysis. We find that light-evoked excitatory postsynaptic currents (L-EPSCs) are mediated by AMPA and N-methyl-d aspartate receptors in IP cells. Although both receptors contributed to the amplitude and kinetics of L-EPSCs, AMPA receptor desensitization substantially shaped L-EPSCs in the neurons, similar to those found in the third-order neurons. The light-evoked inhibitory postsynaptic currents (L-IPSCs) in IP cells were primarily mediated by strychnine-sensitive glycine receptors with a small component of GABA(C) receptors. GABA(C) receptor rho2 subunits were detected in IP cells with single-cell RT-PCR assays. Expression of GABA(C) receptors is one of the special features for IP cells, distinct from most of the third-order neurons that depend on GABA(A) and glycine receptors to relay the inhibitory signals. However, GABA(A) receptors in IP cells acted like nonsynaptic receptors that were activated by exogenous GABA application. Furthermore, L-IPSCs in IP cells were inhibited by the serial inhibitions between amacrine cells in the inner retina. In addition, application of neurotransmitters on the axon terminals of IP cells had no significant current generated in the cells, indicating that the synaptic inputs of IP cells are mainly from the inner retina. This study demonstrates the important role that light signals are encoded by both experiment of inhibitory receptors in IP cells. PMID- 20018831 TI - Spectral and temporal modulation tradeoff in the inferior colliculus. AB - The cochlea encodes sounds through frequency-selective channels that exhibit low pass modulation sensitivity. Unlike the cochlea, neurons in the auditory midbrain are tuned for spectral and temporal modulations found in natural sounds, yet the role of this transformation is not known. We report a distinct tradeoff in modulation sensitivity and tuning that is topographically ordered within the central nucleus of the inferior colliculus (CNIC). Spectrotemporal receptive fields (STRFs) were obtained with 16-channel electrodes inserted orthogonal to the isofrequency lamina. Surprisingly, temporal and spectral characteristics exhibited an opposing relationship along the tonotopic axis. For low best frequencies (BFs), units were selective for fast temporal and broad spectral modulations. A systematic progression was observed toward slower temporal and finer spectral modulation sensitivity at high BF. This tradeoff was strongly reflected in the arrangement of excitation and inhibition and, consequently, in the modulation tuning characteristics. Comparisons with auditory nerve fibers show that these trends oppose the pattern imposed by the peripheral filters. These results suggest that spectrotemporal preferences are reordered within the tonotopic axis of the CNIC. This topographic organization has profound implications for the coding of spectrotemporal features in natural sounds and could underlie a number of perceptual phenomena. PMID- 20018832 TI - Metabotropic glutamate receptors in median preoptic neurons modulate neuronal excitability and glutamatergic and GABAergic inputs from the subfornical organ. AB - Cardiovascular and behavioral responses to circulating angiotensin require intact connectivity along the upper lamina terminalis joining the subfornical organ (SFO) with the median preoptic nucleus (MnPO). In the present study on MnPO neurons, we used whole cell patch-clamp recording techniques in brain slice preparations to evaluate the influence of metabotropic glutamate receptor (mGluR) agonists on modulating their intrinsic excitability and SFO-evoked glutamatergic and GABAergic postsynaptic currents. In 22/36 cells, bath application of a mGluR group I agonist (S)-3,5-dihydroxyphenylglycine (DHPG) induced a TTX-resistant inward current coupled with decrease in a membrane K(+) conductance but also a possible increase in a nonselective cationic conductance. By contrast, 27/49 cells responded to a mGluR group II agonist (2S,2'R,3'R)-2-(2',3' dicarboxycyclopropyl)glycine (DCG IV) with a TTX-resistant outward current and increase in membrane conductance that reversed around -95 mV, suggesting opening of K(+) channels. None of 19 cells responded to the mGluR group III agonist l-(+) 2-amino-4-phosphonobutyric acid (l-AP4). Agonists for all mGluR groups suppressed SFO-evoked excitatory postsynaptic currents and significantly increased paired pulse ratios, implying a presynaptic mechanism. Only the mGluR group II agonist significantly reduced SFO-evoked inhibitory postsynaptic currents and caused an increase in paired-pulse ratios. These results suggest a complexity of pre- and postsynaptic mGluRs are available to modulate rapid neurotransmission along the upper lamina terminalis from SFO to MnPO. PMID- 20018833 TI - Predictive activity in macaque frontal eye field neurons during natural scene searching. AB - Generating sequences of multiple saccadic eye movements allows us to search our environment quickly and efficiently. Although the frontal eye field cortex (FEF) has been linked to target selection and making saccades, little is known about its role in the control and performance of the sequences of saccades made during self-guided visual search. We recorded from FEF cells while monkeys searched for a target embedded in natural scenes and examined the degree to which cells with visual and visuo-movement activity showed evidence of target selection for future saccades. We found that for about half of these cells, activity during the fixation period between saccades predicted the next saccade in a sequence at an early time that precluded selection based on current visual input to a cell's response field. In addition to predicting the next saccade, activity during the fixation prior to two successive saccades also predicted the direction and goal of the second saccade in the sequence. We refer to this as advanced predictive activity. Unlike activity indicating the upcoming saccade, advanced predictive activity occurred later in the fixation period, mirroring the order of the saccade sequence itself. The remaining cells without advanced predictive activity did not predict future saccades but reintroduced the signal for the upcoming saccade at an intermediate time in the fixation period. Together these findings suggest that during natural visual search the timing of FEF cell activity is consistent with a role in specifying targets for one or more future saccades in a search sequence. PMID- 20018834 TI - Changes in the response rate and response variability of area V4 neurons during the preparation of saccadic eye movements. AB - The visually driven responses of macaque area V4 neurons are modulated during the preparation of saccadic eye movements, but the relationship between presaccadic modulation in area V4 and saccade preparation is poorly understood. Recent neurophysiological studies suggest that the variability across trials of spiking responses provides a more reliable signature of motor preparation than mean firing rate across trials. We compared the dynamics of the response rate and the variability in the rate across trials for area V4 neurons during the preparation of visually guided saccades. As in previous reports, we found that the mean firing rate of V4 neurons was enhanced when saccades were prepared to stimuli within a neuron's receptive field (RF) in comparison with saccades to a non-RF location. Further, we found robust decreases in response variability prior to saccades and found that these decreases predicted saccadic reaction times for saccades both to RF and non-RF stimuli. Importantly, response variability predicted reaction time whether or not there were any accompanying changes in mean firing rate. In addition to predicting saccade direction, the mean firing rate could also predict reaction time, but only for saccades directed to the RF stimuli. These results demonstrate that response variability of area V4 neurons, like mean response rate, provides a signature of saccade preparation. However, the two signatures reflect complementary aspects of that preparation. PMID- 20018835 TI - Facilitation of postural limb reflexes with epidural stimulation in spinal rabbits. AB - It is known that after spinalization animals lose their ability to maintain lateral stability when standing or walking. A likely reason for this is a reduction of the postural limb reflexes (PLRs) driven by stretch and load receptors of the limbs. The aim of this study was to clarify whether spinal networks contribute to the generation of PLRs. For this purpose, first, PLRs were recorded in decerebrated rabbits before and after spinalization at T12. Second, the effects of epidural electrical stimulation (EES) at L7 on the limb reflexes were studied after spinalization. To evoke PLRs, the vertebrate column of the rabbit was fixed, whereas the hindlimbs were positioned on the platform. Periodic lateral tilts of the platform caused antiphase flexion-extension limbs movements, similar to those observed in intact animals keeping balance on the tilting platform. Before spinalization, these movements evoked PLRs: augmentation of extensor EMGs and increase of contact force during limb flexion, suggesting their stabilizing postural effects. Spinalization resulted in almost complete disappearance of PLRs. After EES, however, the PLRs reappeared and persisted for up to several minutes, although their values were reduced. The post-EES effects could be magnified by intrathecal application of quipazine (5-HT agonist) at L4 L6. Results of this study suggest that the spinal cord contains the neuronal networks underlying PLRs; they can contribute to the maintenance of lateral stability in intact subjects. In acute spinal animals, these networks can be activated by EES, suggesting that they are normally activated by a tonic supraspinal drive. PMID- 20018836 TI - Tonic GABAA receptor-mediated inhibition in the rat dorsal motor nucleus of the vagus. AB - Type A gamma-aminobutyric acid (GABA(A)) receptors expressed in the dorsal motor nucleus of vagus (DMV) critically regulate the activity of vagal motor neurons and, by inference, the gastrointestinal (GI) tract. Two types of GABA(A) receptor mediated inhibition have been identified in the brain, represented by phasic (I(phasic)) and tonic (I(tonic)) inhibitory currents. The hypothesis that I(tonic) regulates neuron activity was tested in the DMV using whole cell patch clamp recordings in transverse brain stem slices from rats. An I(tonic) was present in a subset of DMV neurons, which was determined to be mediated by different receptors than those mediating fast, synaptic currents. Preapplication of tetrodotoxin significantly decreased the resting I(tonic) amplitude in DMV neurons, suggesting that most of the current was due to action potential (AP) dependent GABA release. Blocking GABA transport enhanced I(tonic) and multiple GABA transporters cooperated to regulate I(tonic). The I(tonic) was composed of both a gabazine-insensitive component that was nearly saturated under basal conditions and a gabazine-sensitive component that was activated when extracellular GABA concentration was elevated. Perfusion of THIP (10 muM) significantly increased I(tonic) amplitude without increasing I(phasic) amplitude. The I(tonic) played a major role in determining the overall excitability of DMV neurons by contributing to resting membrane potential and AP frequency. Our results indicate that I(tonic) contributes to DMV neuron membrane potential and activity and is thus an important regulator of vagally mediated GI function. PMID- 20018837 TI - Roll-dependent modulation of the subjective visual vertical: contributions of head- and trunk-based signals. AB - Precision and accuracy of the subjective visual vertical (SVV) modulate in the roll plane. At large roll angles, systematic SVV errors are biased toward the subject's body-longitudinal axis and SVV precision is decreased. To explain this, SVV models typically implement a bias signal, or a prior, in a head-fixed reference frame and assume the sensory input to be optimally tuned along the head longitudinal axis. We tested the pattern of SVV adjustments both in terms of accuracy and precision in experiments in which the head and the trunk reference frames were not aligned. Twelve subjects were placed on a turntable with the head rolled about 28 degrees counterclockwise relative to the trunk by lateral tilt of the neck to dissociate the orientation of head- and trunk-fixed sensors relative to gravity. Subjects were brought to various positions (roll of head- or trunk longitudinal axis relative to gravity: 0 degrees , +/-75 degrees ) and aligned an arrow with perceived vertical. Both accuracy and precision of the SVV were significantly (P < 0.05) better when the head-longitudinal axis was aligned with gravity. Comparing absolute SVV errors for clockwise and counterclockwise roll tilts, statistical analysis yielded no significant differences (P > 0.05) when referenced relative to head upright, but differed significantly (P < 0.001) when referenced relative to trunk upright. These findings indicate that the bias signal, which drives the SVV toward the subject's body-longitudinal axis, operates in a head-fixed reference frame. Further analysis of SVV precision supports the hypothesis that head-based graviceptive signals provide the predominant input for internal estimates of visual vertical. PMID- 20018838 TI - Vestibuloocular reflex adaptation investigated with chronic motion-modulated electrical stimulation of semicircular canal afferents. AB - To investigate vestibuloocular reflex (VOR) adaptation produced by changes in peripheral vestibular afference, we developed and tested a vestibular "prosthesis" that senses yaw-axis angular head velocity and uses this information to modulate the rate of electrical pulses applied to the lateral canal ampullary nerve. The ability of the brain to adapt the different components of the VOR (gain, phase, axis, and symmetry) during chronic prosthetic electrical stimulation was studied in two squirrel monkeys. After characterizing the normal yaw-axis VOR, electrodes were implanted in both lateral canals and the canals were plugged. The VOR in the canal-plugged/instrumented state was measured and then unilateral stimulation was applied by the prosthesis. The VOR was repeatedly measured over several months while the prosthetic stimulation was cycled between off, low-sensitivity, and high-sensitivity stimulation states. The VOR response initially demonstrated a low gain, abnormal rotational axis, and substantial asymmetry. During chronic stimulation the gain increased, the rotational axis improved, and the VOR became more symmetric. Gain changes were augmented by cycling the stimulation between the off and both low- and high-sensitivity states every few weeks. The VOR time constant remained low throughout the period of chronic stimulation. These results demonstrate that the brain can adaptively modify the gain, axis, and symmetry of the VOR when provided with chronic motion modulated electrical stimulation by a canal prosthesis. PMID- 20018839 TI - Effect of dopaminergic medications on the time course of explicit motor sequence learning in Parkinson's disease. AB - The capacity to learn new motor sequences is fundamental to adaptive motor behavior. The early phase of motor sequence learning relies on the ventral and anterior striatal circuitry, whereas the late phase relies on the dorsal and posterior striatal circuitry. Early Parkinson's disease (PD) is mainly characterized by dopaminergic denervation of the dorsal and posterior striatum while sparing anterior and ventral regions. Dopaminergic medication improves dorsal and posterior striatum function by compensating for the loss of dopamine. However, previous work has shown that dopaminergic medication interferes with the ventral and anterior striatum function by overdosing this relatively intact structure in early-state PD. Here we test whether these effects are also observed over the time course of motor sequence learning. Fourteen PD patients ON and OFF dopaminergic medications and 11 healthy age-matched control participants performed an explicit motor sequence learning task. When sequence learning was compared across different learning phases in patients ON and OFF medication, a significant impairment associated with medication was observed in the early relative to later phases of learning. The rate of learning in the early phase measured trial by trial in patients ON medication was significantly slower than that in controls and when patients were OFF medication. No significant impairment was found in the later learning phases. These results demonstrate that dopaminergic medications may selectively impair early-phase motor sequence learning. These results extend and generalize the dopamine overdose effects previously reported for (antero)ventral striatum-mediated cognitive tasks to motor sequence learning. PMID- 20018840 TI - Identification of the minimal functional unit of the homo-oligomeric human reduced folate carrier. AB - The reduced folate carrier (RFC) is the major transport system for folates in mammals. We previously demonstrated the existence of human RFC (hRFC) homo oligomers and established the importance of these higher order structures to intracellular trafficking and carrier function. In this report, we examined the operational significance of hRFC oligomerization and the minimal functional unit for transport. In negative dominance experiments, multimeric transporters composed of different ratios of active (either wild type (WT) or cysteine-less (CLFL)) and inactive (either inherently inactive (Y281L and R373A) due to mutation, or resulting from inactivation of the Y126C mutant by (2 sulfonatoethyl) methanethiosulfonate (MTSES)) hRFC monomers were expressed in hRFC-null HeLa (R5) cells, and residual WT or CLFL activity was measured. In either case, residual transport activity with increasing levels of inactive mutant correlated linearly with the fraction of WT or CLFL hRFC in plasma membranes. When active covalent hRFC dimers, generated by fusing CLFL and Y126C monomers, were expressed in R5 cells and treated with MTSES, transport activity of the CLFL-CLFL dimer was unaffected, whereas Y126C-Y126C was potently (64%) inhibited; heterodimeric CLFL-Y126C and Y126C-CLFL were only partly (27 and 23%, respectively) inhibited by MTSES. In contrast to Y126C-Y126C, trans-stimulation of methotrexate uptake by intracellular folates for Y126C-CLFL and CLFL-Y126C was nominally affected by MTSES. Collectively, these results strongly support the notion that each hRFC monomer comprises a single translocation pathway for anionic folate substrates and functions independently of other monomers (i.e. despite an oligomeric structure, hRFC functions as a monomer). PMID- 20018841 TI - The C terminus of the Alb3 membrane insertase recruits cpSRP43 to the thylakoid membrane. AB - The YidC/Oxa1/Alb3 family of membrane proteins controls the insertion and assembly of membrane proteins in bacteria, mitochondria, and chloroplasts. Here we describe the molecular mechanisms underlying the interaction of Alb3 with the chloroplast signal recognition particle (cpSRP). The Alb3 C-terminal domain (A3CT) is intrinsically disordered and recruits cpSRP to the thylakoid membrane by a coupled binding and folding mechanism. Two conserved, positively charged motifs reminiscent of chromodomain interaction motifs in histone tails are identified in A3CT that are essential for the Alb3-cpSRP43 interaction. They are absent in the C-terminal domain of Alb4, which therefore does not interact with cpSRP43. Chromodomain 2 in cpSRP43 appears as a central binding platform that can interact simultaneously with A3CT and cpSRP54. The observed negative cooperativity of the two binding events provides the first insights into cargo release at the thylakoid membrane. Taken together, our data show how Alb3 participates in cpSRP-dependent membrane targeting, and our data provide a molecular explanation why Alb4 cannot compensate for the loss of Alb3. Oxa1 and YidC utilize their positively charged, C-terminal domains for ribosome interaction in co-translational targeting. Alb3 is adapted for the chloroplast specific Alb3-cpSRP43 interaction in post-translational targeting by extending the spectrum of chromodomain interactions. PMID- 20018842 TI - Nuclear factor (NF)-kappaB-dependent thyroid hormone receptor beta1 expression controls dendritic cell function via Akt signaling. AB - Despite considerable progress in our understanding of the interplay between immune and endocrine systems, the role of thyroid hormones and their receptors in the control of adaptive immunity is still uncertain. Here, we investigated the role of thyroid hormone receptor (TR) beta(1) signaling in modulating dendritic cell (DC) physiology and the intracellular mechanisms underlying these immunoregulatory effects. Exposure of DCs to triiodothyronine (T(3)) resulted in a rapid and sustained increase in Akt phosphorylation independently of phosphatidylinositol 3-kinase activation, which was essential for supporting T(3) induced DC maturation and interleukin (IL)-12 production. This effect was dependent on intact TR beta(1) signaling as small interfering RNA-mediated silencing of TR beta(1) expression prevented T(3)-induced DC maturation and IL-12 secretion as well as Akt activation and I kappaB-epsilon degradation. In turn, T(3) up-regulated TR beta(1) expression through mechanisms involving NF-kappaB, suggesting an autocrine regulatory loop to control hormone-dependent TR beta(1) signaling. These findings were confirmed by chromatin immunoprecipitation analysis, which disclosed a new functional NF-kappaB consensus site in the promoter region of the TRB1 gene. Thus, a T(3)-induced NF-kappaB-dependent mechanism controls TR beta(1) expression, which in turn signals DCs to promote maturation and function via an Akt-dependent but PI3K-independent pathway. These results underscore a novel unrecognized target that regulates DC maturation and function with critical implications in immunopathology at the cross-roads of the immune-endocrine circuits. PMID- 20018843 TI - Prevention of UVB radiation-induced epidermal damage by expression of heat shock protein 70. AB - Irradiation with UV light, especially UVB, causes epidermal damage via the induction of apoptosis, inflammatory responses, and DNA damage. Various stressors, including UV light, induce heat shock proteins (HSPs) and the induction, particularly that of HSP70, provides cellular resistance to such stressors. The anti-inflammatory activity of HSP70, such as its inhibition of nuclear factor kappa B (NF-kappaB), was recently revealed. These in vitro results suggest that HSP70 protects against UVB-induced epidermal damage. Here we tested this idea by using transgenic mice expressing HSP70 and cultured keratinocytes. Irradiation of wild-type mice with UVB caused epidermal damage such as induction of apoptosis, which was suppressed in transgenic mice expressing HSP70. UVB induced apoptosis in cultured keratinocytes was suppressed by overexpression of HSP70. Irradiation of wild-type mice with UVB decreased the cutaneous level of IkappaB-alpha (an inhibitor of NF-kappaB) and increased the infiltration of leukocytes and levels of pro-inflammatory cytokines and chemokines in the epidermis. These inflammatory responses were suppressed in transgenic mice expressing HSP70. In vitro, the overexpression of HSP70 suppressed the expression of pro-inflammatory cytokines and chemokines and increased the level of IkappaB alpha in keratinocytes irradiated with UVB. UVB induced an increase in cutaneous levels of cyclobutane pyrimidine dimers and 8-hydroxy-2'-deoxyguanosine, both of which were suppressed in transgenic mice expressing HSP70. This study provides genetic evidence that HSP70 protects the epidermis from UVB-induced radiation damage. The findings here also suggest that the protective action of HSP70 is mediated by anti-apoptotic, anti-inflammatory, and anti-DNA damage effects. PMID- 20018844 TI - Growth factor regulation of prostaglandin-endoperoxide synthase 2 (Ptgs2) expression in colonic mesenchymal stem cells. AB - We previously found that a population of colonic stromal cells that constitutively express high levels of prostaglandin-endoperoxide synthase 2 (Ptgs2, also known as Cox-2) altered their location in the lamina propria in response to injury in a Myd88-dependent manner (Brown, S. L., Riehl, T. E., Walker, M. R., Geske, M. J., Doherty, J. M., Stenson, W. F., and Stappenbeck, T. S. (2007) J. Clin. Invest. 117, 258-269). At the time of this study, the identity of these cells and the mechanism by which they expressed high levels of Ptgs2 were unknown. Here we found that these colonic stromal cells were mesenchymal stem cells (MSCs). These colonic MSCs expressed high Ptgs2 levels not through interaction with bacterial products but instead as a consequence of mRNA stabilization downstream of Fgf9 (fibroblast growth factor 9), a growth factor that is constitutively expressed by the intestinal epithelium. This stabilization was mediated partially through a mechanism involving endogenous CUG-binding protein 2 (CUGbp2). These studies suggest that Fgf9 is an important factor in the regulation of Ptgs2 in colonic MSCs and may be a factor involved in its constitutive expression in vivo. PMID- 20018845 TI - CUG start codon generates thioredoxin/glutathione reductase isoforms in mouse testes. AB - Mammalian cytosolic and mitochondrial thioredoxin reductases are essential selenocysteine-containing enzymes that control thioredoxin functions. Thioredoxin/glutathione reductase (TGR) is a third member of this enzyme family. It has an additional glutaredoxin domain and shows highest expression in testes. Herein, we found that human and several other mammalian TGR genes lack any AUG codons that could function in translation initiation. Although mouse and rat TGRs have such codons, we detected protein sequences upstream of them by immunoblot assays and direct proteomic analyses. Further gene engineering and expression analyses demonstrated that a CUG codon, located upstream of the sequences previously thought to initiate translation, is the actual start codon in mouse TGR. The use of this codon relies on the Kozak consensus sequence and ribosome scanning mechanism. However, CUG serves as an inefficient start codon that allows downstream initiation, thus generating two isoforms of the enzyme in vivo and in vitro. The use of CUG evolved in mammalian TGRs, and in some of these organisms, GUG is used instead. The newly discovered longer TGR form shows cytosolic localization in cultured cells and is expressed in spermatids in mouse testes. This study shows that CUG codon is used as an inefficient start codon to generate protein isoforms in mouse. PMID- 20018846 TI - Mys protein regulates protein kinase A activity by interacting with regulatory type Ialpha subunit during vertebrate development. AB - During embryonic development, protein kinase A (PKA) plays a key role in cell fate specification by antagonizing the Hedgehog (Hh) signaling pathway. However, the mechanism by which PKA activity is regulated remains unknown. Here we show that the Misty somites (Mys) protein regulates the level of PKA activity during embryonic development in zebrafish. We isolate PKA regulatory type Ialpha subunit (Prkar1a) as a protein interacting with Mys by pulldown assay in HEK293 cells followed by mass spectrometry analysis. We show an interaction between endogenous Mys and Prkar1a in the zebrafish embryo. Mys binds to Prkar1a in its C terminus region, termed PRB domain, and activates PKA in vitro. Conversely, knockdown of Mys in zebrafish embryos results in reduction in PKA activity. We also show that knockdown of Mys induces ectopic activation of Hh target genes in the eyes, neural tube, and somites downstream of Smoothened, a protein essential for transduction of Hh signaling activity. The altered patterning of gene expression is rescued by activation of PKA. Together, our results reveal a molecular mechanism of regulation of PKA activity that is dependent on a protein-protein interaction and demonstrate that PKA activity regulated by Mys is indispensable for negative regulation of the Hh signaling pathway in Hh-responsive cells. PMID- 20018847 TI - A novel type of E3 ligase for the Ufm1 conjugation system. AB - The ubiquitin fold modifier 1 (Ufm1) is the most recently discovered ubiquitin like modifier whose conjugation (ufmylation) system is conserved in multicellular organisms. Ufm1 is known to covalently attach with cellular protein(s) via a specific E1-activating enzyme (Uba5) and an E2-conjugating enzyme (Ufc1), but its E3-ligating enzyme(s) as well as the target protein(s) remain unknown. Herein, we report both a novel E3 ligase for Ufm1, designated Ufl1, and an Ufm1-specific substrate ligated by Ufl1, C20orf116. Ufm1 was covalently conjugated with C20orf116. Although Ufl1 has no obvious sequence homology to any other known E3s for ubiquitin and ubiquitin-like modifiers, the C20orf116 x Ufm1 formation was greatly accelerated by Ufl1. The C20orf116 x Ufm1 conjugate was cleaved by Ufm1 specific proteases, implying the reversibility of ufmylation. The conjugation was abundant in the liver and lungs of Ufm1-transgenic mice, fractionated into membrane fraction, and impaired in Uba5 knock-out cells. Intriguingly, immunological analysis revealed localizations of Ufl1 and C20orf116 mainly to the endoplasmic reticulum. Our results provide novel insights into the Ufm1 system involved in cellular regulation of multicellular organisms. PMID- 20018848 TI - Azadirachtin interacts with the tumor necrosis factor (TNF) binding domain of its receptors and inhibits TNF-induced biological responses. AB - The role of azadirachtin, an active component of a medicinal plant Neem (Azadirachta indica), on TNF-induced cell signaling in human cell lines was investigated. Azadirachtin blocks TNF-induced activation of nuclear factor kappaB (NF-kappaB) and also expression of NF-kappaB-dependent genes such as adhesion molecules and cyclooxygenase 2. Azadirachtin inhibits the inhibitory subunit of NF-kappaB (IkappaB alpha) phosphorylation and thereby its degradation and RelA (p65) nuclear translocation. It blocks IkappaB alpha kinase (IKK) activity ex vivo, but not in vitro. Surprisingly, azadirachtin blocks NF-kappaB DNA binding activity in transfected cells with TNF receptor-associated factor (TRAF)2, TNF receptor-associated death domain (TRADD), IKK, or p65, but not with TNFR, suggesting its effect is at the TNFR level. Azadirachtin blocks binding of TNF, but not IL-1, IL-4, IL-8, or TNF-related apoptosis-inducing ligand (TRAIL) with its respective receptors. Anti-TNFR antibody or TNF protects azadirachtin mediated down-regulation of TNFRs. Further, in silico data suggest that azadirachtin strongly binds in the TNF binding site of TNFR. Overall, our data suggest that azadirachtin modulates cell surface TNFRs thereby decreasing TNF induced biological responses. Thus, azadirachtin exerts an anti-inflammatory response by a novel pathway, which may be beneficial for anti-inflammatory therapy. PMID- 20018849 TI - JNK pathway-associated phosphatase dephosphorylates focal adhesion kinase and suppresses cell migration. AB - JNK pathway-associated phosphatase (JKAP, also named DUSP22) is expressed in various tissues, indicating that JKAP may have an important biological function. We showed that JKAP localized in the actin filament-enriched region. Expression of JKAP reduced cell migration, whereas a JKAP mutant lacking catalytic activity promoted cell motility. JKAP efficiently removed tyrosine phosphorylation of several proteins. We have identified focal adhesion kinase (FAK) as a substrate of JKAP. Overexpression of JKAP, but not JKAP mutant lacking catalytic activity, decreased FAK phosphorylation at tyrosines 397, 576, and 577 in H1299 cells. Consistent with these results, decreasing JKAP expression by RNA interference promoted cell migration and Src-induced FAK phosphorylation. Taken together, this study identified a new role for JKAP in the modulation of FAK phosphorylation and cell motility. PMID- 20018850 TI - Human alanine-glyoxylate aminotransferase 2 lowers asymmetric dimethylarginine and protects from inhibition of nitric oxide production. AB - Elevated blood concentrations of asymmetric dimethylarginine (ADMA), an endogenous inhibitor of nitric-oxide (NO) synthase, are found in association with diabetes, hypertension, congestive heart failure, and atherosclerosis. ADMA levels are controlled by dimethylarginine dimethylaminohydrolases (DDAHs), cytosolic enzymes that hydrolyze ADMA to citrulline and dimethylamine. ADMA also has been proposed to be regulated through an alternative pathway by alanine glyoxylate aminotransferase 2 (AGXT2), a mitochondrial aminotransferase expressed primarily in the kidney. The goal of this study was to define the subcellular localization of human AGXT2 and test the hypothesis that overexpression of human AGXT2 protects from ADMA-induced inhibition in nitric oxide (NO) production. AGXT2 was cloned from human kidney cDNA and overexpressed in COS-7 cells and human umbilical vein endothelial cells with a C-terminal FLAG epitope tag. Mitochondrial localization of human AGXT2 was demonstrated by confocal microscopy and a 41-amino acid N-terminal mitochondrial cleavage sequence was delineated by N-terminal sequencing of the mature protein. Overexpression of human AGXT2 in the liver of C57BL/6 mice using an adenoviral expression vector produced significant decreases in ADMA levels in plasma and liver. Overexpression of human AGXT2 also protected endothelial cells from ADMA-mediated inhibition of NO production. We conclude that mitochondrially localized human AGXT2 is able to effectively metabolize ADMA in vivo resulting in decreased ADMA levels and improved endothelial NO production. PMID- 20018851 TI - Glucocorticoid-induced leucine zipper (GILZ) promotes the nuclear exclusion of FOXO3 in a Crm1-dependent manner. AB - GILZ (glucocorticoid-induced leucine zipper) is an ubiquitous protein whose expression is induced by glucocorticoids in lymphoid cells. We previously showed that GILZ expression is rapidly induced upon interleukin 2 deprivation in T cells, protecting cells from apoptosis induced by forkhead box subgroup O3 (FOXO3). The aim of this work is to elucidate the molecular mechanism of FOXO factor inhibition by GILZ. We show in the myeloid cell line HL-60 and the lymphoid CTLL-2 T-cell line that GILZ down-regulates the expression of p27(KIP1) and Bim, two FOXO targets involved in cell cycle regulation and apoptosis, respectively. GILZ inhibits FOXO1, FOXO3, and FOXO4 transcriptional activities measured with natural or synthetic FOXO-responsive promoters in HL-60 cells. This inhibitory effect is independent of protein kinase B and IkappaB kinase phosphorylation sites. GILZ does not hinder FOXO3 DNA-binding activity and does not physically interact with FOXO3. However, using fluorescence microscopy, we observe that GILZ expression provokes a Crm-1-dependent nuclear exclusion of FOXO3 leading to its relocalization to the cytoplasm. Moreover, GILZ exclusive cytoplasmic localization is a prerequisite for FOXO3 inhibition and relocalization. We propose that GILZ is a general inhibitor of FOXO factors acting through an original mechanism by preventing them from reaching target genes within the nucleus. PMID- 20018852 TI - Subunit composition and substrate specificity of a MOF-containing histone acetyltransferase distinct from the male-specific lethal (MSL) complex. AB - Human MOF (MYST1), a member of the MYST (Moz-Ybf2/Sas3-Sas2-Tip60) family of histone acetyltransferases (HATs), is the human ortholog of the Drosophila males absent on the first (MOF) protein. MOF is the catalytic subunit of the male specific lethal (MSL) HAT complex, which plays a key role in dosage compensation in the fly and is responsible for a large fraction of histone H4 lysine 16 (H4K16) acetylation in vivo. MOF was recently reported to be a component of a second HAT complex, designated the non-specific lethal (NSL) complex (Mendjan, S., Taipale, M., Kind, J., Holz, H., Gebhardt, P., Schelder, M., Vermeulen, M., Buscaino, A., Duncan, K., Mueller, J., Wilm, M., Stunnenberg, H. G., Saumweber, H., and Akhtar, A. (2006) Mol. Cell 21, 811-823). Here we report an analysis of the subunit composition and substrate specificity of the NSL complex. Proteomic analyses of complexes purified through multiple candidate subunits reveal that NSL is composed of nine subunits. Two of its subunits, WD repeat domain 5 (WDR5) and host cell factor 1 (HCF1), are shared with members of the MLL/SET family of histone H3 lysine 4 (H3K4) methyltransferase complexes, and a third subunit, MCRS1, is shared with the human INO80 chromatin-remodeling complex. In addition, we show that assembly of the MOF HAT into MSL or NSL complexes controls its substrate specificity. Although MSL-associated MOF acetylates nucleosomal histone H4 almost exclusively on lysine 16, NSL-associated MOF exhibits a relaxed specificity and also acetylates nucleosomal histone H4 on lysines 5 and 8. PMID- 20018853 TI - Direct inhibition of Pumilo activity by Bam and Bgcn in Drosophila germ line stem cell differentiation. AB - The fate of stem cells is intricately regulated by numerous extrinsic and intrinsic factors that promote maintenance or differentiation. The RNA-binding translational repressor Pumilio (Pum) in conjunction with Nanos (Nos) is required for self-renewal, whereas Bam (bag-of-marbles) and Bgcn (benign gonial cell neoplasm) promote differentiation of germ line stem cells in the Drosophila ovary. Genetic analysis suggests that Bam and Bgcn antagonize Pum/Nos function to promote differentiation; however, the molecular basis of this epistatic relationship is currently unknown. Here, we show that Bam and Bgcn inhibit Pum function through direct binding. We identified a ternary complex involving Bam, Bgcn, and Pum in which Bam, but not Bgcn, directly interacts with Pum, and this interaction is greatly increased by the presence of Bgcn. In a heterologous reporter assay to monitor Pum activity, Bam, but not Bgcn, inhibits Pum activity. Notably, the N-terminal region of Pum, which lacks the C-terminal RNA-binding Puf domain, mediates both the ternary protein interaction and the Bam inhibition of Pum function. These studies suggest that, in cystoblasts, Bam and Bgcn may directly inhibit Pum/Nos activity to promote differentiation of germ line stem cells. PMID- 20018854 TI - Transcriptional control of human antigen R by bone morphogenetic protein. AB - Human antigen R (HuR) is an RNA-binding protein with protective activities against cellular stress. This study considers the mechanisms by which HuR transcriptional regulation occurs in renal proximal tubule cells. Under basal conditions, HuR mRNA is expressed in two forms: one that contains a approximately 20-base 5'-untranslated region (UTR) sequence and one that contains a approximately 150-base, G+C-rich 5'-UTR that is inhibitory to translation. Recovery from cellular stresses such as thapsigargin and ATP depletion induced increased expression of the shorter, more translatable transcript and decreased expression of the longer form. Analysis of HuR upstream regions revealed sequences necessary for regulation of the shorter mRNA. Within the long, G+C-rich 5'-UTR exist multiple copies of the alternate Smad 1/5/8-binding motif GCCGnCGC. Recovery from ATP depletion induced increases in Smad 1/5/8 levels; further, gel shift and chromatin immunoprecipitation analyses demonstrated the ability of these Smads to bind to the relevant motif in the HuR 5'-UTR. Transfection of exogenous Smad 1 increased HuR mRNA expression. Finally, HuR mRNA expression driven by the Smad-binding sites was responsive to BMP-7, a protein with known protective effects against ischemic injury in kidney. These data suggest that transcriptional induction of a readily translatable HuR mRNA may be driven by a mechanism known to protect the kidney from injury and provides a novel pathway through which administration of BMP-7 may attenuate renal damage. PMID- 20018855 TI - Cross-species binding analyses of mouse and human neonatal Fc receptor show dramatic differences in immunoglobulin G and albumin binding. AB - The neonatal Fc receptor (FcRn) regulates the serum half-life of both IgG and albumin through a pH-dependent mechanism that involves salvage from intracellular degradation. Therapeutics and diagnostics built on IgG, Fc, and albumin fusions are frequently evaluated in rodents regarding biodistribution and pharmacokinetics. Thus, it is important to address cross-species ligand reactivity with FcRn, because in vivo testing of such molecules is done in the presence of competing murine ligands, both in wild type (WT) and human FcRn (hFcRn) transgenic mice. Here, binding studies were performed in vitro using enzyme-linked immunosorbent assay and surface plasmon resonance with recombinant soluble forms of human (shFcRn(WT)) and mouse (smFcRn(WT)) receptors. No binding of albumin from either species was observed at physiological pH to either receptor. At acidic pH, a 100-fold difference in binding affinity was observed. Specifically, smFcRn(WT) bound human serum albumin with a K(D) of approximately 90 microM, whereas shFcRn(WT) bound mouse serum albumin with a K(D) of 0.8 microM. shFcRn(WT) ignored mouse IgG1, and smFcRn(WT) bound strongly to human IgG1. The latter pair also interacted at physiological pH with calculated affinity in the micromolar range. In all cases, binding of albumin and IgG from either species to both receptors were additive. Cross-species albumin binding differences could partly be explained by non-conserved amino acids found within the alpha2-domain of the receptor. Such distinct cross-species FcRn binding differences must be taken into consideration when IgG- and albumin-based therapeutics and diagnostics are evaluated in rodents for their pharmacokinetics. PMID- 20018856 TI - Involvement of the spliceosomal U4 small nuclear RNA in heterochromatic gene silencing at fission yeast centromeres. AB - prp13-1 is one of the mutants isolated in a screen for defective pre-mRNA splicing at a nonpermissive temperature in fission yeast Schizosaccharomyces pombe. We cloned the prp13(+) gene and found that it encodes U4 small nuclear RNA (snRNA) involved in the assembly of the spliceosome. The prp13-1 mutant produced elongated cells, a phenotype similar to cell division cycle mutants, and displays a high incidence of lagging chromosomes on anaphase spindles. The mutant is hypersensitive to the microtubule-destabilizing drug thiabendazole, supporting that prp13-1 has a defect in chromosomal segregation. We found that the prp13-1 mutation resulted in expression of the ura4(+) gene inserted in the pericentromeric heterochromatin region and reduced recruitment of the heterochromatin protein Swi6p to that region, indicating defects in the formation of pericentromeric heterochromatin, which is essential for the segregation of chromosomes, in prp13-1. The formation of centromeric heterochromatin is induced by the RNA interference (RNAi) system in S. pombe. In prp13-1, the processing of centromeric noncoding RNAs to siRNAs, which direct the heterochromatin formation, was impaired and unprocessed noncoding RNAs were accumulated. These results suggest that U4 snRNA is required for the RNAi-directed heterochromatic gene silencing at the centromeres. In relation to the linkage between the spliceosomal U4 snRNA and the RNAi-directed formation of heterochromatin, we identified a mRNA type intron in the centromeric noncoding RNAs. We propose a model in which the assembly of the spliceosome or a sub-spliceosome complex on the intron-containing centromeric noncoding RNAs facilitates the RNAi-directed formation of heterochromatin at centromeres, through interaction with the RNA-directed RNA polymerase complex. PMID- 20018858 TI - Myosin light chain kinase is necessary for tonic airway smooth muscle contraction. AB - Different interacting signaling modules involving Ca(2+)/calmodulin-dependent myosin light chain kinase, Ca(2+)-independent regulatory light chain phosphorylation, myosin phosphatase inhibition, and actin filament-based proteins are proposed as specific cellular mechanisms involved in the regulation of smooth muscle contraction. However, the relative importance of specific modules is not well defined. By using tamoxifen-activated and smooth muscle-specific knock-out of myosin light chain kinase in mice, we analyzed its role in tonic airway smooth muscle contraction. Knock-out of the kinase in both tracheal and bronchial smooth muscle significantly reduced contraction and myosin phosphorylation responses to K(+)-depolarization and acetylcholine. Kinase-deficient mice lacked bronchial constrictions in normal and asthmatic airways, whereas the asthmatic inflammation response was not affected. These results indicate that myosin light chain kinase acts as a central participant in the contractile signaling module of tonic smooth muscle. Importantly, contractile airway smooth muscles are necessary for physiological and asthmatic airway resistance. PMID- 20018857 TI - Cyclic mechanical stretch decreases cell migration by inhibiting phosphatidylinositol 3-kinase- and focal adhesion kinase-mediated JNK1 activation. AB - Epithelial cell migration during wound healing requires coordinated signaling pathways that direct polarization of the leading and trailing ends of the cells, cytoskeletal organization, and remodeling of focal adhesions. These inherently mechanical processes are disrupted by cyclic stretch (CS), but the specific signaling molecules involved in this disruption are not well understood. In this study, we demonstrate that inhibition of phosphatidylinositol 3-kinase (PI3K) or expression of a dominant-negative form of PI3K caused inhibition of airway epithelial cell wound closure. CS caused a sustained decrease in activation of PI3K and inhibited wound healing. Expression of constitutively active PI3K stimulated translocation of Tiam1 to the membrane, increased Rac1 activity, and increased wound healing of airway epithelial cells. Increased Rac1 activity resulted in increased phosphorylation of JNK1. PI3K activation was not regulated by association with focal adhesion kinase. Restoration of efficient cell migration during CS required coexpression of constitutively active PI3K, focal adhesion kinase, and JIP3. PMID- 20018859 TI - The neuropeptide calcitonin gene-related peptide causes repression of tumor necrosis factor-alpha transcription and suppression of ATF-2 promoter recruitment in Toll-like receptor-stimulated dendritic cells. AB - Sensory nerves may dampen inflammatory processes through the release of the neuropeptide calcitonin gene-related peptide (CGRP). CGRP mediates immunosuppressive activities through up-regulation of interleukin-10 or, alternatively, through an interleukin-10-independent pathway that is associated with rapid induction of the transcriptional inducible cAMP early repressor (ICER). In this work, we further investigated the molecular mechanisms of immune modulation by CGRP. Using TLR2-stimulated dendritic cells, we show that inhibition of tumor necrosis factor-alpha production by CGRP is dependent on up regulation of endogenous ICER. Dendritic cell expression of ICER was selectively induced by CGRP and elevation of cellular cAMP levels but not by numerous pro- and anti-inflammatory cytokines. Treatment of dendritic cells with CGRP did not interfere with the induction of Tnfa gene expression but caused premature repression of TLR2-induced transcriptional activity. ATF-2 was rapidly phosphorylated and recruited to the Tnfa promoter following ligation of TLR2. Concomitant administration of CGRP completely prevented binding of ATF-2 to the Tnfa promoter, whereas recruitment of ICER was markedly elevated. In contrast, CGRP did not influence TLR2-stimulated binding of NF-kappaB p65. Together, these results are consistent with a model suggesting that CGRP causes rapid up regulation of ICER, which in turn competes with ATF-2 for binding to the Tnfa promoter, leading to repression of gene expression. PMID- 20018860 TI - A protein-protein interaction map of trypanosome ~20S editosomes. AB - Mitochondrial mRNA editing in trypanosomatid parasites involves several multiprotein assemblies, including three very similar complexes that contain the key enzymatic editing activities and sediment at ~20S on glycerol gradients. These ~20S editosomes have a common set of 12 proteins, including enzymes for uridylyl (U) removal and addition, 2 RNA ligases, 2 proteins with RNase III-like domains, and 6 proteins with predicted oligonucleotide binding (OB) folds. In addition, each of the 3 distinct ~20S editosomes contains a different RNase III type endonuclease, 1 of 3 related proteins and, in one case, an additional exonuclease. Here we present a protein-protein interaction map that was obtained through a combination of yeast two-hybrid analysis and subcomplex reconstitution with recombinant protein. This map interlinks ten of the proteins and in several cases localizes the protein region mediating the interaction, which often includes the predicted OB-fold domain. The results indicate that the OB-fold proteins form an extensive protein-protein interaction network that connects the two trimeric subcomplexes that catalyze U removal or addition and RNA ligation. One of these proteins, KREPA6, interacts with the OB-fold zinc finger protein in each subcomplex that interconnects their two catalytic proteins. Another OB-fold protein, KREPA3, appears to link to the putative endonuclease subcomplex. These results reveal a physical organization that underlies the coordination of the various catalytic and substrate binding activities within the ~20S editosomes during the editing process. PMID- 20018861 TI - Human GTSE-1 regulates p21(CIP1/WAF1) stability conferring resistance to paclitaxel treatment. AB - p21(CIP1/WAF1) belongs to the CIP/KIP family of Cdk inhibitors, and its expression is tightly controlled during the cell cycle, mainly by transcriptional and post-translational mechanisms. Fine regulation of p21(CIP1/WAF1) levels is critical for cell cycle control and for cellular response to stress. In the present work, we describe a novel mechanism to modulate p21(CIP1/WAF1) levels mediated by the human GTSE-1 (G(2) and S phase-expressed-1) protein. Our results provide evidence that hGTSE-1 protects p21(CIP1/WAF1) from proteasome-dependent degradation as part of a functional complex containing the Hsp90-binding TPR protein WISp39. We further show that the hGTSE-1 N-terminal portion is sufficient for p21(CIP1/WAF1) binding and stabilization. Finally, we demonstrate that hGTSE 1 mediated-p21(CIP1/WAF1) stabilization is clearly involved in the ability of cells to counteract cytotoxicity induced by the microtubule poison paclitaxel. PMID- 20018862 TI - NF-kappaB functions in stromal fibroblasts to regulate early postnatal muscle development. AB - Classical NF-kappaB activity functions as an inhibitor of the skeletal muscle myogenic program. Recent findings reveal that even in newborn RelA/p65(-/-) mice, myofiber numbers are increased over that of wild type mice, suggesting that NF kappaB may be a contributing factor in early postnatal skeletal muscle development. Here we show that in addition to p65 deficiency, repression of NF kappaB with the IkappaB alpha-SR transdominant inhibitor or with muscle-specific deletion of IKKbeta resulted in similar increases in total fiber numbers as well as an up-regulation of myogenic gene products. Upon further characterization of early postnatal muscle, we observed that NF-kappaB activity progressively declines within the first few weeks of development. At birth, the majority of this activity is compartmentalized to muscle fibers, but by neonatal day 8 NF kappaB activity from the myofibers diminishes, and instead, stromal fibroblasts become the main cellular compartment within the muscle that contains active NF kappaB. We find that NF-kappaB functions in these fibroblasts to regulate inducible nitric-oxide synthase expression, which we show is important for myoblast fusion during the growth and maturation process of skeletal muscle. Together, these data broaden our understanding of NF-kappaB during development by showing that in addition to its role as a negative regulator of myogenesis, NF kappaB also regulates nitric-oxide synthase expression within stromal fibroblasts to stimulate myoblast fusion and muscle hypertrophy. PMID- 20018863 TI - Real-time NMR study of three small GTPases reveals that fluorescent 2'(3')-O-(N methylanthraniloyl)-tagged nucleotides alter hydrolysis and exchange kinetics. AB - The Ras family of small GTPases control diverse signaling pathways through a conserved "switch" mechanism, which is turned on by binding of GTP and turned off by GTP hydrolysis to GDP. Full understanding of GTPase switch functions requires reliable, quantitative assays for nucleotide binding and hydrolysis. Fluorescently labeled guanine nucleotides, such as 2'(3')-O-(N methylanthraniloyl) (mant)-substituted GTP and GDP analogs, have been widely used to investigate the molecular properties of small GTPases, including Ras and Rho. Using a recently developed NMR method, we show that the kinetics of nucleotide hydrolysis and exchange by three small GTPases, alone and in the presence of their cognate GTPase-activating proteins (GAPs) and guanine nucleotide exchange factors, are affected by the presence of the fluorescent mant moiety. Intrinsic hydrolysis of mantGTP by Ras homolog enriched in brain (Rheb) is approximately 10 times faster than that of GTP, whereas it is 3.4 times slower with RhoA. On the other hand, the mant tag inhibits TSC2GAP-catalyzed GTP hydrolysis by Rheb but promotes p120 RasGAP-catalyzed GTP hydrolysis by H-Ras. Guanine nucleotide exchange factor-catalyzed nucleotide exchange for both H-Ras and RhoA was inhibited by mant-substituted nucleotides, and the degree of inhibition depends highly on the GTPase and whether the assay measures association of mantGTP with, or dissociation of mantGDP from the GTPase. These results indicate that the mant moiety has significant and unpredictable effects on GTPase reaction kinetics and underscore the importance of validating its use in each assay. PMID- 20018864 TI - Neuronal calcium sensor-1 (Ncs1p) is up-regulated by calcineurin to promote Ca2+ tolerance in fission yeast. AB - Neuronal calcium sensor (NCS) proteins regulate signal transduction and are highly conserved from yeast to humans. NCS homolog in fission yeast (Ncs1p) is essential for cell growth under extreme Ca(2+) conditions. Ncs1p expression increases approximately 100-fold when fission yeast grows in high extracellular Ca(2+) (>0.1 M). Here, we show that Ca(2+)-induced expression of Ncs1p is controlled at the level of transcription. Transcriptional reporter assays show that ncs1 promoter activity increased 30-fold when extracellular Ca(2+) was raised to 0.1 M and was highly Ca(2+)-specific. Ca(2+)-dependent transcription of ncs1 is abolished by the calcineurin inhibitor (FK506) and by knocking out the calcineurin target, prz1. Thus, Ca(2+)-induced expression of Ncs1p is linked to the calcineurin/prz1 stress response. The Ca(2+)-responsive ncs1 promoter region consists of 130 nucleotides directly upstream from the start codon and contains tandem repeats of the sequence, 5'-caact-3', that binds to Prz1p. The Ca(2+) sensitive ncs1Delta phenotype is rescued by a yam8 null mutation, suggesting a possible interaction between Ncs1p and the Ca(2+) channel, Yam8p. Ca(2+) uptake and Ncs1p binding to yeast membranes are both decreased in yam8Delta, suggesting Ca(2+)-induced binding of Ncs1p to Yam8p results in channel closure. We propose that Ncs1p promotes Ca(2+) tolerance in fission yeast, in part by cytosolic Ca(2+) buffering and perhaps by negatively regulating the Yam8p Ca(2+) channel. PMID- 20018865 TI - Uncoupling of inflammation and insulin resistance by NF-kappaB in transgenic mice through elevated energy expenditure. AB - To study the metabolic activity of NF-kappaB, we investigated phenotypes of two different mouse models with elevated NF-kappaB activities. The transcriptional activity of NF-kappaB is enhanced either by overexpression of NF-kappaB p65 (RelA) in aP2-p65 mice or inactivation of NF-kappaB p50 (NF-kappaB1) through gene knock-out. In these models, energy expenditure was elevated in day and night time without a change in locomotion. The mice were resistant to adulthood obesity and diet-induced obesity without reduction in food intake. The adipose tissue growth and adipogenesis were inhibited by the elevated NF-kappaB activity. Peroxisome proliferator-activator receptor gamma expression was reduced by NF-kappaB at the transcriptional level. The two models exhibited elevated inflammatory cytokines (tumor necrosis factor-alpha and interleukin-6) in adipose tissue and serum. However, insulin sensitivity was not reduced by the inflammation in the mice on a chow diet. On a high fat diet, the mice were protected from insulin resistance. The glucose infusion rate was increased more than 30% in the hyperinsulinemic euglycemic clamp test. Our data suggest that the transcription factor NF-kappaB promotes energy expenditure and inhibits adipose tissue growth. The two effects lead to prevention of adulthood obesity and dietary obesity. The energy expenditure may lead to disassociation of inflammation with insulin resistance. The study indicates that inflammation may prevent insulin resistance by eliminating lipid accumulation. PMID- 20018866 TI - Akt and c-Myc differentially activate cellular metabolic programs and prime cells to bioenergetic inhibition. AB - The high glucose consumption of tumor cells even in an oxygen-rich environment, referred to as the Warburg effect, has been noted as a nearly universal biochemical characteristic of cancer cells. Targeting the glycolysis pathway has been explored as an anti-cancer therapeutic strategy to eradicate cancer based on this fundamental biochemical property of cancer cells. Oncoproteins such as Akt and c-Myc regulate cell metabolism. Accumulating studies have uncovered various molecular mechanisms by which oncoproteins affect cellular metabolism, raising a concern as to whether targeting glycolysis will be equally effective in treating cancers arising from different oncogenic activities. Here, we established a dual regulatable FL5.12 pre-B cell line in which myristoylated Akt is expressed under the control of doxycycline, and c-Myc, fused to the hormone-binding domain of the human estrogen receptor, is activated by 4-hydroxytamoxifen. Using this system, we directly compared the effect of these oncoproteins on cell metabolism in an isogenic background. Activation of either Akt or c-Myc leads to the Warburg effect as indicated by increased cellular glucose uptake, glycolysis, and lactate generation. When cells are treated with glycolysis inhibitors, Akt sensitizes cells to apoptosis, whereas c-Myc does not. In contrast, c-Myc but not Akt sensitizes cells to the inhibition of mitochondrial function. This is correlated with enhanced mitochondrial activities in c-Myc cells. Hence, although both Akt and c-Myc promote aerobic glycolysis, they differentially affect mitochondrial functions and render cells susceptible to the perturbation of cellular metabolic programs. PMID- 20018867 TI - Reactive oxygen generated by NADPH oxidase 1 (Nox1) contributes to cell invasion by regulating matrix metalloprotease-9 production and cell migration. AB - A mediating role of the reactive oxygen species-generating enzyme Nox1 has been suggested for Ras oncogene transformation phenotypes including anchorage independent cell growth, augmented angiogenesis, and tumorigenesis. However, little is known about whether Nox1 signaling regulates cell invasiveness. Here, we report that the cell invasion activity was augmented in K-Ras-transformed normal rat kidney cells and attenuated by transfection of Nox1 small interference RNAs (siRNAs) into the cells. Diphenyleneiodonium (DPI) or Nox1 siRNAs blocked up regulation of matrix metalloprotease-9 at both protein and mRNA levels in K-Ras transformed normal rat kidney cells. Furthermore, DPI and Nox1 siRNAs inhibited the activation of IKKalpha kinase and the degradation of IkappaB alpha, suppressing the NFkappaB-dependent matrix metalloprotease-9 promoter activity. Additionally, epidermal growth factor-stimulated migration of CaCO-2 cells was abolished by DPI and Nox1 siRNAs, indicating the requirement of Nox1 activity for the motogenic effect of epidermal growth factor. This Nox1 action was mediated by down-regulation of the Rho activity through the low molecular weight protein tyrosine phosphatase-p190RhoGAP-dependent mechanism. Taken together, our findings define a mediating role of Nox1-generated reactive oxygen species in cell invasion processes, most notably metalloprotease production and cell motile activity. PMID- 20018868 TI - Regulation of insulin receptor substrate 1 (IRS-1)/AKT kinase-mediated insulin signaling by O-Linked beta-N-acetylglucosamine in 3T3-L1 adipocytes. AB - Increased O-linked beta-N-acetylglucosamine (O-GlcNAc) is associated with insulin resistance in muscle and adipocytes. Upon insulin treatment of insulin-responsive adipocytes, O-GlcNAcylation of several proteins is increased. Key insulin signaling proteins, including IRS-1, IRS-2, and PDK1, are substrates for OGT, suggesting potential O-GlcNAc control points within the pathway. To elucidate the roles of O-GlcNAc in dampening insulin signaling (Vosseller, K., Wells, L., Lane, M. D., and Hart, G. W. (2002) Proc. Natl. Acad. Sci. U. S. A. 99, 5313-5318), we focused on the pathway upstream of AKT. Increasing O-GlcNAc in 3T3-L1 adipocytes decreases phosphoinositide 3-kinase (PI3K) interactions with both IRS-1 and IRS 2. Elevated O-GlcNAc also reduces phosphorylation of the PI3K p85 binding motifs (YXXM) of IRS-1 and results in a concomitant reduction in tyrosine phosphorylation of Y(608)XXM in IRS-1, one of the two main PI3K p85 binding motifs. Additionally, insulin signaling stimulates the interaction of OGT with PDK1. We conclude that one of the steps at which O-GlcNAc contributes to insulin resistance is by inhibiting phosphorylation at the Y(608)XXM PI3K p85 binding motif in IRS-1 and possibly at PDK1 as well. PMID- 20018869 TI - Real-time NMR study of guanine nucleotide exchange and activation of RhoA by PDZ RhoGEF. AB - Small guanosine triphosphatases (GTPases) become activated when GDP is replaced by GTP at the highly conserved nucleotide binding site. This process is intrinsically very slow in most GTPases but is significantly accelerated by guanine nucleotide exchange factors (GEFs). Nucleotide exchange in small GTPases has been widely studied using spectroscopy with fluorescently tagged nucleotides. However, this method suffers from effects of the bulky fluorescent moiety covalently attached to the nucleotide. Here, we have used a newly developed real time NMR-based assay to monitor small GTPase RhoA nucleotide exchange by probing the RhoA conformation. We compared RhoA nucleotide exchange from GDP to GTP and GTP analogues in the absence and presence of the catalytic DH-PH domain of PDZ RhoGEF (DH-PH(PRG)). Using the non-hydrolyzable analogue guanosine-5'-O-(3 thiotriphosphate), which we found to be a reliable mimic of GTP, we obtained an intrinsic nucleotide exchange rate of 5.5 x 10(-4) min(-1). This reaction is markedly accelerated to 1179 x 10(-4) min(-1) in the presence of DH-PH(PRG) at a ratio of 1:8,000 relative to RhoA. Mutagenesis studies confirmed the importance of Arg-868 near a conserved region (CR3) of the Dbl homology (DH) domain and revealed that Glu-741 in CR1 is critical for full activity of DH-PH(PRG), together suggesting that the catalytic mechanism of PDZ-RhoGEF is similar to Tiam1. Mutation of the single RhoA (E97A) residue that contacts the pleckstrin homology (PH) domain rendered the mutant 10-fold less sensitive to the activity of DH-PH(PRG). Interestingly, this mutation does not affect RhoA activation by leukemia-associated RhoGEF (LARG), indicating that the PH domains of these two homologous GEFs may play different roles. PMID- 20018871 TI - Sulfation of colonic mucins by N-acetylglucosamine 6-O-sulfotransferase-2 and its protective function in experimental colitis in mice. AB - N-Acetylglucosamine 6-O-sulfotransferase-2 (GlcNAc6ST-2) catalyzes the sulfation of mucin-like glycoproteins, which function as ligands for a lymphocyte homing receptor, L-selectin, in the lymph node high endothelial venules (HEVs). We previously showed that GlcNAc6ST-2 is expressed not only in lymph node HEVs but also in the colonic epithelial cells in mice. Here we investigated the regulatory mechanism and physiological significance of colonic expression of GlcNAc6ST-2 in mice. Treatment of a mouse colonic epithelial cell line with butyrate, a short chain fatty acid produced by anaerobic bacteria, induced GlcNAc6ST-2 expression in the presence of epidermal growth factor. Administration of butyrate in the drinking water stimulated GlcNAc6ST-2 expression in the mouse intestine, indicating that butyrate could serve as a regulatory molecule for the GlcNAc6ST-2 expression in vivo. Immunohistochemical analysis indicated that the sulfation of colonic mucins was greatly diminished in GlcNAc6ST-2-deficient mice. Liquid chromatography coupled to electrospray ionization tandem mass spectrometry of the colonic-mucin O-glycans from wild-type and GlcNAc6ST-2-deficient mice showed that GlcNAc-6-O-sulfation was the predominant sulfate modification of these mucins, and it was exclusively mediated by GlcNAc6ST-2. After colitis induction by dextran sulfate sodium, significantly more leukocyte infiltration was observed in the colon of GlcNAc6ST-2-deficient mice than in that of wild-type mice, indicating that the sulfation of colonic mucins by GlcNAc6ST-2 has a protective function in experimental colitis. These findings indicate that GlcNAc6ST-2, whose expression is regulated by butyrate, is a major sulfotransferase in the biosynthesis of sulfomucins in the mouse colon, where they serve as a mucosal barrier against colonic inflammation. PMID- 20018870 TI - Distinct sarcomeric substrates are responsible for protein kinase D-mediated regulation of cardiac myofilament Ca2+ sensitivity and cross-bridge cycling. AB - Protein kinase D (PKD), a serine/threonine kinase with emerging cardiovascular functions, phosphorylates cardiac troponin I (cTnI) at Ser(22)/Ser(23), reduces myofilament Ca(2+) sensitivity, and accelerates cross-bridge cycle kinetics. Whether PKD regulates cardiac myofilament function entirely through cTnI phosphorylation at Ser(22)/Ser(23) remains to be established. To determine the role of cTnI phosphorylation at Ser(22)/Ser(23) in PKD-mediated regulation of cardiac myofilament function, we used transgenic mice that express cTnI in which Ser(22)/Ser(23) are substituted by nonphosphorylatable Ala (cTnI-Ala(2)). In skinned myocardium from wild-type (WT) mice, PKD increased cTnI phosphorylation at Ser(22)/Ser(23) and decreased the Ca(2+) sensitivity of force. In contrast, PKD had no effect on the Ca(2+) sensitivity of force in myocardium from cTnI Ala(2) mice, in which Ser(22)/Ser(23) were unavailable for phosphorylation. Surprisingly, PKD accelerated cross-bridge cycle kinetics similarly in myocardium from WT and cTnI-Ala(2) mice. Because cardiac myosin-binding protein C (cMyBP-C) phosphorylation underlies cAMP-dependent protein kinase (PKA)-mediated acceleration of cross-bridge cycle kinetics, we explored whether PKD phosphorylates cMyBP-C at its PKA sites, using recombinant C1C2 fragments with or without site-specific Ser/Ala substitutions. Kinase assays confirmed that PKA phosphorylates Ser(273), Ser(282), and Ser(302), and revealed that PKD phosphorylates only Ser(302). Furthermore, PKD phosphorylated Ser(302) selectively and to a similar extent in native cMyBP-C of skinned myocardium from WT and cTnI-Ala(2) mice, and this phosphorylation occurred throughout the C-zones of sarcomeric A-bands. In conclusion, PKD reduces myofilament Ca(2+) sensitivity through cTnI phosphorylation at Ser(22)/Ser(23) but accelerates cross-bridge cycle kinetics by a distinct mechanism. PKD phosphorylates cMyBP-C at Ser(302), which may mediate the latter effect. PMID- 20018873 TI - Hepatocyte nuclear factor-4alpha induces transdifferentiation of hematopoietic cells into hepatocytes. AB - Hematopoietic stem cells can directly transdifferentiate into hepatocytes because of cellular plasticity, but the molecular basis of transdifferentiation is not known. Here, we show the molecular basis using lineage-depleted oncostatin M receptor beta-expressing (Lin(-)OSMRbeta(+)) mouse bone marrow cells in a hepatic differentiation culture system. Differentiation of the cells was marked by the expression of albumin. Hepatocyte nuclear factor (HNF)-4alpha was expressed and translocated into the nuclei of the differentiating cells. Suppression of its activation in OSM-neutralized culture medium inhibited cellular differentiation. Ectopic expression of full-length HNF4alpha in 32D myeloid cells resulted in decreased myeloid colony-forming potential and increased expression of hepatocyte specific genes and proteins. Nevertheless, the neohepatocytes produced in culture expressed active P450 enzyme. The obligatory role of HNF4alpha in hepatic differentiation was confirmed by transfecting Lin(-)OSMRbeta(+) cells with dominant negative HNF4alpha in the differentiation culture because its expression inhibited the transcription of the albumin and tyrosine aminotransferase genes. The loss and gain of functional activities strongly suggested that HNF4alpha plays a central role in the transdifferentiation process. For the first time, this report demonstrates the mechanism of transdifferentiation of hematopoietic cells into hepatocytes, in which HNF4alpha serves as a molecular switch. PMID- 20018872 TI - FoxO proteins mediate hypoxic induction of connective tissue growth factor in endothelial cells. AB - Hypoxia, a driving force in neovascularization, promotes alterations in gene expression mediated by hypoxia-inducible factor (HIF)-1alpha. Connective tissue growth factor (CTGF, CCN2) is a modulator of endothelial cell growth and migration, but its regulation by hypoxia is poorly understood. Therefore, we analyzed signaling pathways involved in the regulation of CTGF by hypoxia in endothelial cells. Exposure to low oxygen tension or treatment with the hypoxia mimetic dimethyloxalyl glycine (DMOG) stabilized HIF-1alpha and up-regulated CTGF in human umbilical vein endothelial cells and in a murine microvascular endothelial cell line. Induction of CTGF correlated with a HIF-dependent increase in protein and mRNA levels, and nuclear accumulation of the transcription factor FoxO3a. By contrast, gene expression and cellular localization of FoxO1 were not significantly altered by hypoxia. Expression of CTGF was strongly reduced by siRNA silencing of FoxO1 or FoxO3a. Furthermore, nuclear exclusion of FoxO1/3a transcription factors by inhibition of serine/threonine protein phosphatases by okadaic acid inhibited CTGF expression, providing evidence for both FoxO proteins as regulators of CTGF expression. The DMOG-stimulated induction of CTGF was further increased when endothelial cells were co-incubated with transforming growth factor-beta, an activator of Smad signaling. Activation of RhoA-Rho kinase signaling by the microtubule-disrupting drug combretastatin A4 also enhanced the DMOG-induced CTGF expression, thus placing CTGF induction by hypoxia in a network of interacting signaling pathways. Our findings provide evidence that FoxO1, hypoxia-stimulated expression of FoxO3a and its nuclear accumulation are required for the induction of CTGF by hypoxia in endothelial cells. PMID- 20018874 TI - Wnt2 regulates progenitor proliferation in the developing ventral midbrain. AB - Wnts are secreted, lipidated proteins that regulate multiple aspects of brain development, including dopaminergic neuron development. In this study, we perform the first purification and signaling analysis of Wnt2 and define the function of Wnt2 in ventral midbrain precursor cultures, as well as in Wnt2-null mice in vivo. We found that purified Wnt2 induces the phosphorylation of both Lrp5/6 and Dvl-2/3, and activates beta-catenin in SN4741 dopaminergic cells. Moreover, purified Wnt2 increases progenitor proliferation, and the number of dopaminergic neurons in ventral midbrain precursor cultures. In agreement with these findings, analysis of the ventral midbrain of developing Wnt2-null mice revealed a decrease in progenitor proliferation and neurogenesis that lead to a decrease in the number of postmitotic precursors and dopaminergic neurons. Collectively, our observations identify Wnt2 as a novel regulator of dopaminergic progenitors and dopaminergic neuron development. PMID- 20018875 TI - G alpha(i) and G betagamma jointly regulate the conformations of a G betagamma effector, the neuronal G protein-activated K+ channel (GIRK). AB - Stable complexes among G proteins and effectors are an emerging concept in cell signaling. The prototypical G betagamma effector G protein-activated K(+) channel (GIRK; Kir3) physically interacts with G betagamma but also with G alpha(i/o). Whether and how G alpha(i/o) subunits regulate GIRK in vivo is unclear. We studied triple interactions among GIRK subunits 1 and 2, G alpha(i3) and G betagamma. We used in vitro protein interaction assays and in vivo intramolecular Forster resonance energy transfer (i-FRET) between fluorophores attached to N and C termini of either GIRK1 or GIRK2 subunit. We demonstrate, for the first time, that G betagamma and G alpha(i3) distinctly and interdependently alter the conformational states of the heterotetrameric GIRK1/2 channel. Biochemical experiments show that G betagamma greatly enhances the binding of GIRK1 subunit to G alpha(i3)(GDP) and, unexpectedly, to G alpha(i3)(GTP). i-FRET showed that both G alpha(i3) and G betagamma induced distinct conformational changes in GIRK1 and GIRK2. Moreover, GIRK1 and GIRK2 subunits assumed unique, distinct conformations when coexpressed with a "constitutively active" G alpha(i3) mutant and G betagamma together. These conformations differ from those assumed by GIRK1 or GIRK2 after separate coexpression of either G alpha(i3) or G betagamma. Both biochemical and i-FRET data suggest that GIRK acts as the nucleator of the GIRK-G alpha-G betagamma signaling complex and mediates allosteric interactions between G alpha(i)(GTP) and G betagamma. Our findings imply that G alpha(i/o) and the G alpha(i) betagamma heterotrimer can regulate a G betagamma effector both before and after activation by neurotransmitters. PMID- 20018876 TI - Aquaporin-1 tunes pain perception by interaction with Na(v)1.8 Na+ channels in dorsal root ganglion neurons. AB - Aquaporin-1 (AQP1) water channels are expressed in the plasma membrane of dorsal root ganglion (DRG) neurons. We found reduced osmotic water permeability in freshly isolated DRG neurons from AQP1(-/-) versus AQP1(+/+) mice. Behavioral studies showed greatly reduced thermal inflammatory pain perception in AQP1(-/-) mice evoked by bradykinin, prostaglandin E(2), and capsaicin as well as reduced cold pain perception. Patch clamp of freshly isolated DRG neurons showed reduced action potential firing in response to current injections. Single action potentials after pulse current injections showed reduced maximum inward current, suggesting impaired Na(v)1.8 Na(+) function. Whole-cell Na(v)1.8 Na(+) currents in Na(v)1.8-expressing ND7-23 cells showed slowed frequency-dependent inactivation after AQP1 transfection. Immunoprecipitation studies showed AQP1- Na(v)1.8 Na(+) interaction, which was verified in live cells by single-particle tracking of quantum dot-labeled AQP1. Our results implicate the involvement of AQP1 in DRG neurons for the perception of inflammatory thermal pain and cold pain, whose molecular basis is accounted for, in part, by reduced Na(v)1.8 dependent membrane Na(+) current. AQP1 is, thus, a novel target for pain management. PMID- 20018877 TI - Crystal structure and computational analyses provide insights into the catalytic mechanism of 2,4-diacetylphloroglucinol hydrolase PhlG from Pseudomonas fluorescens. AB - 2,4-Diacetylphloroglucinol hydrolase PhlG from Pseudomonas fluorescens catalyzes hydrolytic carbon-carbon (C-C) bond cleavage of the antibiotic 2,4 diacetylphloroglucinol to form monoacetylphloroglucinol, a rare class of reactions in chemistry and biochemistry. To investigate the catalytic mechanism of this enzyme, we determined the three-dimensional structure of PhlG at 2.0 A resolution using x-ray crystallography and MAD methods. The overall structure includes a small N-terminal domain mainly involved in dimerization and a C terminal domain of Bet v1-like fold, which distinguishes PhlG from the classical alpha/beta-fold hydrolases. A dumbbell-shaped substrate access tunnel was identified to connect a narrow interior amphiphilic pocket to the exterior solvent. The tunnel is likely to undergo a significant conformational change upon substrate binding to the active site. Structural analysis coupled with computational docking studies, site-directed mutagenesis, and enzyme activity analysis revealed that cleavage of the 2,4-diacetylphloroglucinol C-C bond proceeds via nucleophilic attack by a water molecule, which is coordinated by a zinc ion. In addition, residues Tyr(121), Tyr(229), and Asn(132), which are predicted to be hydrogen-bonded to the hydroxyl groups and unhydrolyzed acetyl group, can finely tune and position the bound substrate in a reactive orientation. Taken together, these results revealed the active sites and zinc dependent hydrolytic mechanism of PhlG and explained its substrate specificity as well. PMID- 20018878 TI - Mechanism and role of high density lipoprotein-induced activation of AMP activated protein kinase in endothelial cells. AB - The upstream signaling pathway leading to the activation of AMP-activated protein kinase (AMPK) by high density lipoprotein (HDL) and the role of AMPK in HDL induced antiatherogenic actions were investigated. Experiments using genetic and pharmacological tools showed that HDL-induced activation of AMPK is dependent on both sphingosine 1-phosphate receptors and scavenger receptor class B type I through calcium/calmodulin-dependent protein kinase kinase and, for scavenger receptor class B type I system, additionally serine-threonine kinase LKB1 in human umbilical vein endothelial cells. HDL-induced activation of Akt and endothelial NO synthase, stimulation of migration, and inhibition of monocyte adhesion and adhesion molecule expression were dependent on AMPK activation. The inhibitory role of AMPK in the adhesion molecule expression and monocyte adhesion on endothelium of mouse aorta was confirmed in vivo and ex vivo. On the other hand, stimulation of ERK and proliferation were hardly affected by AMPK knockdown but completely inhibited by an N17Ras, whereas the dominant-negative Ras was ineffective for AMPK activation. In conclusion, dual HDL receptor systems differentially regulate AMPK activity through calcium/calmodulin-dependent protein kinase kinase and/or LKB1. Several HDL-induced antiatherogenic actions are regulated by AMPK, but proliferation-related actions are regulated by Ras rather than AMPK. PMID- 20018879 TI - Slow onset inhibition of bacterial beta-ketoacyl-acyl carrier protein synthases by thiolactomycin. AB - Thiolactomycin (TLM), a natural product thiolactone antibiotic produced by species of Nocardia and Streptomyces, is an inhibitor of the beta-ketoacyl-acyl carrier protein synthase (KAS) enzymes in the bacterial fatty acid synthase pathway. Using enzyme kinetics and direct binding studies, TLM has been shown to bind preferentially to the acyl-enzyme intermediates of the KASI and KASII enzymes from Mycobacterium tuberculosis and Escherichia coli. These studies, which utilized acyl-enzyme mimics in which the active site cysteine was replaced by a glutamine, also revealed that TLM is a slow onset inhibitor of the KASI enzymes KasA and ecFabB but not of the KASII enzymes KasB and ecFabF. The differential affinity of TLM for the acyl-KAS enzymes is proposed to result from structural change involving the movement of helices alpha5 and alpha6 that prepare the enzyme to bind malonyl-AcpM or TLM and that is initiated by formation of hydrogen bonds between the acyl-enzyme thioester and the oxyanion hole. The finding that TLM is a slow onset inhibitor of ecFabB supports the proposal that the long residence time of TLM on the ecFabB homologues in Serratia marcescens and Klebsiella pneumonia is an important factor for the in vivo antibacterial activity of TLM against these two organisms despite the fact that the in vitro MIC values are only 100-200 microg/ml. The mechanistic data on the interaction of TLM with KasA will provide an important foundation for the rational development of high affinity KasA inhibitors based on the thiolactone skeleton. PMID- 20018880 TI - Cross-talk between remodeling and de novo pathways maintains phospholipid balance through ubiquitination. AB - Phosphatidylcholine (PtdCho), the major phospholipid of animal membranes, is generated by its remodeling and de novo synthesis. Overexpression of the remodeling enzyme, LPCAT1 (acyl-CoA:lysophosphatidylcholine acyltransferase) in epithelia decreased de novo PtdCho synthesis without significantly altering cellular PtdCho mass. Overexpression of LPCAT1 increased degradation of CPT1 (cholinephosphotransferase), a resident Golgi enzyme that catalyzes the terminal step for de novo PtdCho synthesis. CPT1 degradation involved its multiubiquitination and processing via the lysosomal pathway. CPT1 mutants harboring arginine substitutions at multiple carboxyl-terminal lysines exhibited proteolytic resistance to effects of LPCAT1 overexpression in cells and restored de novo PtdCho synthesis. Thus, cross-talk between phospholipid remodeling and de novo pathways involves ubiquitin-lysosomal processing of a key molecular target that mechanistically provides homeostatic control of cellular PtdCho content. PMID- 20018881 TI - Gap filling activities of Pseudomonas DNA ligase D (LigD) polymerase and functional interactions of LigD with the DNA end-binding Ku protein. AB - Many bacterial pathogens, including Pseudomonas aeruginosa, have a nonhomologous end joining (NHEJ) system of DNA double strand break (DSB) repair driven by Ku and DNA ligase D (LigD). LigD is a multifunctional enzyme composed of a ligase domain fused to an autonomous polymerase module (POL) that adds ribonucleotides or deoxyribonucleotides to DSB ends and primer-templates. LigD POL and the eukaryal NHEJ polymerase lambda are thought to bridge broken DNA ends via contacts with a duplex DNA segment downstream of the primer terminus, a scenario analogous to gap repair. Here, we characterized the gap repair activity of Pseudomonas LigD POL, which is more efficient than simple templated primer extension and relies on a 5'-phosphate group on the distal gap strand end to confer apparent processivity in filling gaps of 3 or 4 nucleotides. Mutations of the His-553, Arg-556, and Lys-566 side chains implicated in DNA 5'-phosphate binding eliminate the preferential filling of 5'-phosphate gaps. Mutating Phe 603, which is imputed to stack on the nucleobase of the template strand that includes the 1st bp of the downstream gap duplex segment, selectively affects incorporation of the final gap-closing nucleotide. We find that Pseudomonas Ku stimulates POL-catalyzed ribonucleotide addition to a plasmid DSB end and promotes plasmid end joining by full-length Pseudomonas LigD. A series of incremental truncations from the C terminus of the 293-amino acid Ku polypeptide identifies Ku-(1-229) as sufficient for homodimerization and LigD stimulation. The slightly longer Ku-(1-253) homodimer forms stable complexes at both ends of linear plasmid DNA that protect the DSBs from digestion by 5'- and 3' exonucleases. PMID- 20018882 TI - Replication initiation at a distance: determination of the cis- and trans-acting elements of replication origin alpha of plasmid R6K. AB - Plasmid R6K, which contains 3 replication origins called alpha, gamma, and beta, is a favorable system to investigate the molecular mechanism(s) of action at a distance, i.e. replication initiation at a considerable distance from the primary initiator protein binding sites (iterons). The centrally located gamma origin contains 7 iterons that bind to the plasmid-encoded initiator protein, pi. Ori alpha, located at a distance of approximately 4 kb from gamma, contains a single iteron that does not directly bind to pi but is believed to access the protein by pi-mediated alpha-gamma iteron-iteron interaction that loops out the intervening approximately 3.7 kb of DNA. Although the cis-acting components and the trans acting proteins required for ori gamma function have been analyzed in detail, such information was lacking for ori alpha. Here, we have identified both the sequence elements located at alpha and those at gamma, that together promoted alpha activity. The data support the conclusion that besides the single iteron, a neighboring DNA primase recognition element called G site is essential for alpha directed plasmid maintenance. Sequences preceding the iteron and immediately following the G site, although not absolutely necessary, appear to play a role in efficient plasmid maintenance. In addition, while both dnaA1 and dnaA2 boxes that bind to DnaA protein and are located at gamma were essential for alpha activity, only dnaA2 was required for initiation at gamma. Mutations in the AT-rich region of gamma also abolished alpha function. These results are consistent with the interpretation that a protein-DNA complex consisting of pi and DnaA forms at gamma and activates alpha at a distance by DNA looping. PMID- 20018883 TI - Processing of the matricellular protein hevin in mouse brain is dependent on ADAMTS4. AB - The matricellular SPARC family member hevin (SPARC-like 1/SPARCL-1/SC1/Mast9) contributes to neural development and alters tumor progression in a range of mammalian models. The distribution of hevin in mouse tissues was reexamined with a novel monoclonal antibody that discriminates between hevin and its ortholog SPARC. We now report proteolysis of hevin in many tissues, with the most extensive processing in the brain. We demonstrate a cleavage site within the hevin sequence for the neural tissue proteinase ADAMTS4. Digestion of hevin by ADAMTS4 in vitro produced fragments similar to those present in brain lysates. Monoclonal antibodies revealed a SPARC-like fragment generated from hevin that was co-localized with ADAMTS4 in vivo. We show that proteolysis of hevin by ADAMTS4 in the mouse cerebellum is important for the normal development of this tissue. In conclusion, we have identified the fragmentation of hevin by ADAMTS4 in the mouse brain and propose that this specific proteolysis is integral to cell morphology and extracellular matrix deposition in the developing brain. PMID- 20018884 TI - An experimentally based computer search identifies unstructured membrane-binding sites in proteins: application to class I myosins, PAKS, and CARMIL. AB - Programs exist for searching protein sequences for potential membrane-penetrating segments (hydrophobic regions) and for lipid-binding sites with highly defined tertiary structures, such as PH, FERM, C2, ENTH, and other domains. However, a rapidly growing number of membrane-associated proteins (including cytoskeletal proteins, kinases, GTP-binding proteins, and their effectors) bind lipids through less structured regions. Here, we describe the development and testing of a simple computer search program that identifies unstructured potential membrane binding sites. Initially, we found that both basic and hydrophobic amino acids, irrespective of sequence, contribute to the binding to acidic phospholipid vesicles of synthetic peptides that correspond to the putative membrane-binding domains of Acanthamoeba class I myosins. Based on these results, we modified a hydrophobicity scale giving Arg- and Lys-positive, rather than negative, values. Using this basic and hydrophobic scale with a standard search algorithm, we successfully identified previously determined unstructured membrane-binding sites in all 16 proteins tested. Importantly, basic and hydrophobic searches identified previously unknown potential membrane-binding sites in class I myosins, PAKs and CARMIL (capping protein, Arp2/3, myosin I linker; a membrane-associated cytoskeletal scaffold protein), and synthetic peptides and protein domains containing these newly identified sites bound to acidic phospholipids in vitro. PMID- 20018885 TI - Nucleocytoplasmic shuttling of p62/SQSTM1 and its role in recruitment of nuclear polyubiquitinated proteins to promyelocytic leukemia bodies. AB - p62, also known as sequestosome1 (SQSTM1), A170, or ZIP, is a multifunctional protein implicated in several signal transduction pathways. p62 is induced by various forms of cellular stress, is degraded by autophagy, and acts as a cargo receptor for autophagic degradation of ubiquitinated targets. It is also suggested to shuttle ubiquitinated proteins for proteasomal degradation. p62 is commonly found in cytosolic protein inclusions in patients with protein aggregopathies, it is up-regulated in several forms of human tumors, and mutations in the gene are linked to classical adult onset Paget disease of the bone. To this end, p62 has generally been considered to be a cytosolic protein, and little attention has been paid to possible nuclear roles of this protein. Here, we present evidence that p62 shuttles continuously between nuclear and cytosolic compartments at a high rate. The protein is also found in nuclear promyelocytic leukemia bodies. We show that p62 contains two nuclear localization signals and a nuclear export signal. Our data suggest that the nucleocytoplasmic shuttling of p62 is modulated by phosphorylations at or near the most important nuclear localization signal, NLS2. The aggregation of p62 in cytosolic bodies also regulates the transport of p62 between the compartments. We found p62 to be essential for accumulation of polyubiquitinated proteins in promyelocytic leukemia bodies upon inhibition of nuclear protein export. Furthermore, p62 contributed to the assembly of proteasome-containing degradative compartments in the vicinity of nuclear aggregates containing polyglutamine-expanded Ataxin1Q84 and to the degradation of Ataxin1Q84. PMID- 20018886 TI - Characterization of Streptococcus pyogenes beta-NAD+ glycohydrolase: re evaluation of enzymatic properties associated with pathogenesis. AB - The gram-positive pathogen Streptococcus pyogenes injects a beta-NAD(+) glycohydrolase (SPN) into the cytosol of an infected host cell using the cytolysin-mediated translocation pathway. In this compartment, SPN accelerates the death of the host cell by an unknown mechanism that may involve its beta NAD(+)-dependent enzyme activities. SPN has been reported to possess the unique characteristic of not only catalyzing hydrolysis of beta-NAD(+), but also carrying out ADP-ribosyl cyclase and ADP-ribosyltransferase activities, making SPN the only beta-NAD(+) glycohydrolase that can catalyze all of these reactions. With the long term goal of understanding how these activities may contribute to pathogenesis, we have further characterized the enzymatic activity of SPN using highly purified recombinant protein. Kinetic studies of the multiple activities of SPN revealed that SPN possessed only beta-NAD(+) hydrolytic activity and lacked detectable ADP-ribosyl cyclase and ADP-ribosyltransferase activities. Similarly, SPN was unable to catalyze cyclic ADPR hydrolysis, and could not catalyze methanolysis or transglycosidation. Kinetic analysis of product inhibition by recombinant SPN demonstrated an ordered uni-bi mechanism, with ADP ribose being released as a second product. SPN was unaffected by product inhibition using nicotinamide, suggesting that this moiety contributes little to the binding energy of the substrate. Upon transformation, SPN was toxic to Saccharomyces cerevisiae, whereas a glycohydrolase-inactive SPN allowed for viability. Taken together, these data suggest that SPN functions exclusively as a strict beta-NAD(+) glycohydrolase during pathogenesis. PMID- 20018887 TI - Phosphorylation of maize eukaryotic translation initiation factor 5A (eIF5A) by casein kinase 2: identification of phosphorylated residue and influence on intracellular localization of eIF5A. AB - Maize eukaryotic translation initiation factor 5A (ZmeIF5A) co-purifies with the catalytic alpha subunit of protein kinase CK2 and is phosphorylated by this enzyme. Phosphorylated ZmeIF5A was also identified after separation of maize leaf proteins by two-dimensional electrophoresis. Multiple sequence alignment of eIF5A proteins showed that in monocots, in contrast to other eukaryotes, there are two serine/threonine residues that could potentially be phosphorylated by CK2. To identify the phosphorylation site(s) of ZmeIF5A, the serine residues potentially phosphorylated by CK2 were mutated. ZmeIF5A and its mutated variants S2A and S4A were expressed in Escherichia coli and purified. Of these recombinant proteins, only ZmeIF5A-S2A was not phosphorylated by maize CK2. Also, Arabidopsis thaliana and Saccharomyces cerevisiae eIF5A-S2A mutants were not phosphorylated despite effective phosphorylation of wild-type variants. A newly developed method exploiting the specificity of thrombin cleavage was used to confirm that Ser(2) in ZmeIF5A is indeed phosphorylated. To find a role of the Ser(2) phosphorylation, ZmeIF5A and its variants mutated at Ser(2) (S2A and S2D) were transiently expressed in maize protoplasts. The expressed fluorescence labeled proteins were visualized by confocal microscopy. Although wild-type ZmeIF5A and its S2A variant were distributed evenly between the nucleus and cytoplasm, the variant with Ser(2) replaced by aspartic acid, which mimics a phosphorylated serine, was sequestered in the nucleus. These results suggests that phosphorylation of Ser(2) plays a role in regulation of nucleocytoplasmic shuttling of eIF5A in plant cells. PMID- 20018888 TI - Overcoming amino-Nogo-induced inhibition of cell spreading and neurite outgrowth by 12-O-tetradecanoylphorbol-13-acetate-type tumor promoters. AB - The N-terminal domain of NogoA, called amino-Nogo, inhibits axonal outgrowth and cell spreading via a largely unknown mechanism. In the present study, we show that amino-Nogo decreases Rac1 activity and inhibits fibroblast spreading. 12-O Tetradecanoylphorbol-13-acetate-type tumor promoters, such as phorbol 12 myristate 13-acetate (PMA) and teleocidin, increase Rac1 activity and overcome the amino-Nogo-induced inhibition of cell spreading. The stimulating effect of tumor promoters on cell spreading requires activation of protein kinase D and the subsequent activation of Akt1. Furthermore, we identified Akt1 as a new signaling component of the amino-Nogo pathway. Akt1 phosphorylation is decreased by amino Nogo. Activation of Akt1 with a cell-permeable peptide, TAT-TCL1, blocks the amino-Nogo inhibition. Finally, we provide evidence that these signaling pathways operate in neurons in addition to fibroblasts. Our results suggest that activation of protein kinase D and Akt1 are approaches to promote axonal regeneration after injury. PMID- 20018889 TI - Fibrillar oligomers nucleate the oligomerization of monomeric amyloid beta but do not seed fibril formation. AB - Soluble amyloid oligomers are potent neurotoxins that are involved in a wide range of human degenerative diseases, including Alzheimer disease. In Alzheimer disease, amyloid beta (Abeta) oligomers bind to neuronal synapses, inhibit long term potentiation, and induce cell death. Recent evidence indicates that several immunologically distinct structural variants exist as follows: prefibrillar oligomers (PFOs), fibrillar oligomers (FOs), and annular protofibrils. Despite widespread interest, amyloid oligomers are poorly characterized in terms of structural differences and pathological significance. FOs are immunologically related to fibrils because they react with OC, a conformation-dependent, fibril specific antibody and do not react with antibodies specific for other types of oligomers. However, fibrillar oligomers are much smaller than fibrils. FOs are soluble at 100,000 x g, rich in beta-sheet structures, but yet bind weakly to thioflavin T. EPR spectroscopy indicates that FOs display significantly more spin spin interaction at multiple labeled sites than PFOs and are more structurally similar to fibrils. Atomic force microscopy indicates that FOs are approximately one-half to one-third the height of mature fibrils. We found that Abeta FOs do not seed the formation of thioflavin T-positive fibrils from Abeta monomers but instead seed the formation of FOs from Abeta monomers that are positive for the OC anti-fibril antibody. These results indicate that the lattice of FOs is distinct from the fibril lattice even though the polypeptide chains are organized in an immunologically identical conformation. The FOs resulting from seeded reactions have the same dimensions and morphology as the initial seeds, suggesting that the seeds replicate by growing to a limiting size and then splitting, indicating that their lattice is less stable than fibrils. We suggest that FOs may represent small pieces of single fibril protofilament and that the addition of monomers to the ends of FOs is kinetically more favorable than the assembly of the oligomers into fibrils via sheet stacking interaction. These studies provide novel structural insight into the relationship between fibrils and FOs and suggest that the increased toxicity of FOs may be due to their ability to replicate and the exposure of hydrophobic sheet surfaces that are otherwise obscured by sheet-sheet interactions between protofilaments in a fibril. PMID- 20018890 TI - SSeCKS/Gravin/AKAP12 inhibits cancer cell invasiveness and chemotaxis by suppressing a protein kinase C- Raf/MEK/ERK pathway. AB - SSeCKS/Gravin/AKAP12 ("SSeCKS") encodes a cytoskeletal protein that regulates G(1) --> S progression by scaffolding cyclins, protein kinase C (PKC) and PKA. SSeCKS is down-regulated in many tumor types including prostate, and when re expressed in MAT-LyLu (MLL) prostate cancer cells, SSeCKS selectively inhibits metastasis by suppressing neovascularization at distal sites, correlating with its ability to down-regulate proangiogenic genes including Vegfa. However, the forced re-expression of VEGF only rescues partial lung metastasis formation. Here, we show that SSeCKS potently inhibits chemotaxis and Matrigel invasion, motility parameters contributing to metastasis formation. SSeCKS suppressed serum induced activation of the Raf/MEK/ERK pathway, resulting in down-regulation of matrix metalloproteinase-2 expression. In contrast, SSeCKS had no effect on serum induced phosphorylation of the Src substrate, Shc, in agreement with our previous data that SSeCKS does not inhibit Src kinase activity in cells. Invasiveness and chemotaxis could be restored by the forced expression of constitutively active MEK1, MEK2, ERK1, or PKCalpha. SSeCKS suppressed phorbol ester-induced ERK1/2 activity only if it encoded its PKC binding domain (amino acids 553-900), suggesting that SSeCKS attenuates ERK activation through a direct scaffolding of conventional and/or novel PKC isozymes. Finally, control of MLL invasiveness by SSeCKS is influenced by the actin cytoskeleton: the ability of SSeCKS to inhibit podosome formation is unaffected by cytochalasin D or jasplakinolide, whereas its ability to inhibit MEK1/2 and ERK1/2 activation is nullified by jasplakinolide. Our findings suggest that SSeCKS suppresses metastatic motility by disengaging activated Src and then inhibiting the PKC-Raf/MEK/ERK pathways controlling matrix metalloproteinase-2 expression and podosome formation. PMID- 20018891 TI - GSK-3 represses growth factor-inducible genes by inhibiting NF-kappaB in quiescent cells. AB - GSK-3 is active in the absence of growth factor stimulation and generally acts to induce apoptosis or inhibit cell proliferation. We previously identified a subset of growth factor-inducible genes that can also be induced in quiescent T98G cells solely by inhibition of GSK-3 in the absence of growth factor stimulation. Computational predictions verified by chromatin immunoprecipitation assays identified NF-kappaB binding sites in the upstream regions of 75% of the genes regulated by GSK-3. p50 bound to most of these sites in quiescent cells, and for one-third of the genes, binding of p65 to the predicted sites increased upon inhibition of GSK-3. The functional role of p65 in gene induction following inhibition of GSK-3 was demonstrated by RNA interference experiments. Furthermore, inhibition of GSK-3 in quiescent cells resulted in activation of IkappaB kinase, leading to phosphorylation and degradation of IkappaB alpha and nuclear translocation of p65 and p50. Taken together, these results indicate that the high levels of GSK-3 activity in quiescent cells repress gene expression by negatively regulating NF-kappaB through inhibition of IkappaB kinase. This inhibition of NF-kappaB is consistent with the role of GSK-3 in the induction of apoptosis or cell cycle arrest in cells deprived of growth factors. PMID- 20018892 TI - The structure of calreticulin C-terminal domain is modulated by physiological variations of calcium concentration. AB - Calreticulin is an abundant endoplasmic reticulum resident protein that fulfills at least two basic functions. Firstly, due to its ability to bind monoglucosylated high mannose oligosaccharides, calreticulin is a central component of the folding quality control system of glycoproteins. On the other hand, thanks to its capacity to bind high amounts of calcium, calreticulin is one of the main calcium buffers in the endoplasmic reticulum. This last activity resides on a highly negatively charged domain located at the C terminus. Interestingly, this domain has been proposed to regulate the intracellular localization of calreticulin. Structural information for this domain is currently scarce. Here we address this issue by employing a combination of biophysical techniques and molecular dynamics simulation. We found that calreticulin C terminal domain at low calcium concentration displays a disordered structure, whereas calcium addition induces a more rigid and compact conformation. Remarkably, this change develops when calcium concentration varies within a range similar to that taking place in the endoplasmic reticulum upon physiological fluctuations. In addition, a much higher calcium concentration is necessary to attain similar responses in a peptide displaying a randomized sequence of calreticulin C-terminal domain, illustrating the sequence specificity of this effect. Molecular dynamics simulation reveals that this ordering effect is a consequence of the ability of calcium to bring into close proximity residues that lie apart in the primary structure. These results place calreticulin in a new setting in which the protein behaves not only as a calcium-binding protein but as a finely tuned calcium sensor. PMID- 20018893 TI - MD-2-mediated ionic interactions between lipid A and TLR4 are essential for receptor activation. AB - Lipopolysaccharide (LPS) activates innate immune responses through TLR4.MD-2. LPS binds to the MD-2 hydrophobic pocket and bridges the dimerization of two TLR4.MD 2 complexes to activate intracellular signaling. However, exactly how lipid A, the endotoxic moiety of LPS, activates myeloid lineage cells remains unknown. Lipid IV(A), a tetra-acylated lipid A precursor, has been used widely as a model for lipid A activation. For unknown reasons, lipid IV(A) activates proinflammatory responses in rodent cells but inhibits the activity of LPS in human cells. Using stable TLR4-expressing cell lines and purified monomeric MD-2, as well as MD-2-deficient bone marrow-derived macrophages, we found that both mouse TLR4 and mouse MD-2 are required for lipid IV(A) activation. Computational studies suggested that unique ionic interactions exist between lipid IV(A) and TLR4 at the dimerization interface in the mouse complex only. The negatively charged 4'-phosphate on lipid IV(A) interacts with two positively charged residues on the opposing mouse, but not human, TLR4 (Lys(367) and Arg(434)) at the dimerization interface. When replaced with their negatively charged human counterparts Glu(369) and Gln(436), mouse TLR4 was no longer responsive to lipid IV(A). In contrast, human TLR4 gained lipid IV(A) responsiveness when ionic interactions were enabled by charge reversal at the dimerization interface, defining the basis of lipid IV(A) species specificity. Thus, using lipid IV(A) as a selective lipid A agonist, we successfully decoupled and coupled two sequential events required for intracellular signaling: receptor engagement and dimerization, underscoring the functional role of ionic interactions in receptor activation. PMID- 20018894 TI - MicroRNAs regulate human hepatocyte nuclear factor 4alpha, modulating the expression of metabolic enzymes and cell cycle. AB - Hepatocyte nuclear factor (HNF) 4alpha is a key transcription factor regulating endo/xenobiotic-metabolizing enzymes and transporters. We investigated whether microRNAs are involved in the regulation of human HNF4alpha. Potential recognition elements for miR-24 (MRE24) were identified in the coding region and the 3'-untranslated region (3'-UTR), and those for miR-34a (MRE34a) were identified in the 3'-UTR in HNF4alpha mRNA. The HNF4alpha protein level in HepG2 cells was markedly decreased by the overexpression of miR-24 and miR-34a. The HNF4alpha mRNA level was significantly decreased by the overexpression of miR-24 but not by miR-34a. In luciferase analyses in HEK293 cells, the reporter activity of plasmid containing the 3'-UTR of HNF4alpha was significantly decreased by miR 34a. The reporter activity of plasmid containing the HNF4alpha coding region downstream of the luciferase gene was significantly decreased by miR-24. These results suggest that the MRE24 in the coding region and MRE34a in the 3'-UTR are functional in the negative regulation by mRNA degradation and translational repression, respectively. The down-regulation of HNF4alpha by these microRNAs resulted in the decrease of various target genes such as cytochrome P450 7A1 and 8B1 as well as morphological changes and the decrease of the S phase population in HepG2 cells. We also clarified that the expressions of miR-24 and miR-34a were regulated by protein kinase C/mitogen-activated protein kinase and reactive oxygen species pathways, respectively. In conclusion, we found that human HNF4alpha was down-regulated by miR-24 and miR-34a, the expression of which are regulated by cellular stress, affecting the metabolism and cellular biology. PMID- 20018895 TI - FGF19-induced hepatocyte proliferation is mediated through FGFR4 activation. AB - FGF19 and FGF21, unique members of the fibroblast growth factor (FGF) family, are hormones that regulate glucose, lipid, and energy homeostasis. Increased hepatocyte proliferation and liver tumor formation have also been observed in FGF19 transgenic mice. Here, we report that, in contrast to FGF19, FGF21 does not induce hepatocyte proliferation in vivo. To identify the mechanism for FGF19 induced hepatocyte proliferation, we explored similarities and differences in receptor specificity between FGF19 and FGF21. We find that although both are able to activate FGF receptors (FGFRs) 1c, 2c, and 3c, only FGF19 activates FGFR4, the predominant receptor in the liver. Using a C-terminal truncation mutant of FGF19 and a series of FGF19/FGF21 chimeric molecules, we determined that amino acids residues 38-42 of FGF19 are sufficient to confer both FGFR4 activation and increased hepatocyte proliferation in vivo to FGF21. These data suggest that activation of FGFR4 is the mechanism whereby FGF19 can increase hepatocyte proliferation and induce hepatocellular carcinoma formation. PMID- 20018896 TI - Recruitment of cAMP-response element-binding protein and histone deacetylase has opposite effects on glucocorticoid receptor gene transcription. AB - Glucocorticoids control the synthesis of the glucocorticoid receptor (GR) in various tissues through a negative feedback regulation of the mRNA. In this study, we have identified feedback regulatory domains in the human GR gene promoter and examined the roles of GR, the cAMP-response element-binding protein (CREB), and HDAC-6 in association with promoter elements of the human GR gene. Using breast cancer T47D and HeLa-GR cells, we identify specific negative glucocorticoid-response elements in the GR gene. The feedback regulatory domains were also involved in interactions with CREB. GR-bound negative glucocorticoid response elements recruited HDAC-6, and this was dependent on treatment with dexamethasone. Both CREB and HDAC-6 formed complexes with GR-dexamethasone. The HDAC-6 LXXLL motif between amino acids 313 and 418 made direct contact with the GR AF-1 domain. Interestingly enough, although the level of GR decreased in CREB knockdown cells, it was elevated in HDAC-6 knockdown cells. Our results suggest that CREB-P is dephosphorylated and that HDAC-6 is recruited by the GR, and they play opposite roles in the negative feedback regulation of the GR gene. PMID- 20018897 TI - ATP hydrolysis in Eg5 kinesin involves a catalytic two-water mechanism. AB - Motor proteins couple steps in ATP binding and hydrolysis to conformational switching both in and remote from the active site. In our kinesin.AMPPPNP crystal structure, closure of the active site results in structural transformations appropriate for microtubule binding and organizes an orthosteric two-water cluster. We conclude that a proton is shared between the lytic water, positioned for gamma-phosphate attack, and a second water that serves as a general base. To our knowledge, this is the first experimental detection of the catalytic base for any ATPase. Deprotonation of the second water by switch residues likely triggers subsequent large scale structural rearrangements. Therefore, the catalytic base is responsible for initiating nucleophilic attack of ATP and for relaying the positive charge over long distances to initiate mechanotransduction. Coordination of switch movements via sequential proton transfer along paired water clusters may be universal for nucleotide triphosphatases with conserved active sites, such as myosins and G-proteins. PMID- 20018898 TI - Annexin A2 is a molecular target for TM601, a peptide with tumor-targeting and anti-angiogenic effects. AB - TM601 is a synthetic form of chlorotoxin, a 36-amino acid peptide derived from the venom of the Israeli scorpion, Leirius quinquestriatus, initially found to specifically bind and inhibit the migration of glioma cells in culture. Subsequent studies demonstrated specific in vitro binding to additional tumor cell lines. Recently, we demonstrated that proliferating human vascular endothelial cells are the only normal cell line tested that exhibits specific binding to TM601. Here, we identify annexin A2 as a novel binding partner for TM601 in multiple human tumor cell lines and human umbilical vein endothelial cell (HUVEC). We demonstrate that the surface binding of TM601 to the pancreatic tumor cell line Panc-1 is dependent on the expression of annexin A2. Identification of annexin A2 as a binding partner for TM601 is also consistent with the anti-angiogenic effects of TM601. Annexin A2 functions in angiogenesis by binding to tissue plasminogen activator and regulating plasminogen activation on vascular endothelial cells. We demonstrate that in HUVECs, TM601 inhibits both vascular endothelial growth factor- and basic fibroblast growth factor-induced tissue plasminogen activator activation, which is required for activation of plasminogen to plasmin. Consistent with inhibition of cell surface protease activity, TM601 also inhibits platelet-derived growth factor-C induced trans-well migration of both HUVEC and U373-MG glioma cells. PMID- 20018900 TI - Nicotiana tabacum agglutinin is active against Lepidopteran pest insects. AB - A jasmonate-inducible lectin called Nicotiana tabacum agglutinin or NICTABA was found in tobacco (Nicotiana tabacum cv Samsun) leaves. Since NICTABA expression is also induced after insect herbivory, a role in the defence response of tobacco was suggested. In this report, a detailed analysis was made of the entomotoxic properties of NICTABA using different transgenic approaches. First, purified NICTABA was shown to be strongly resistant to proteolytic degradation by enzymes present in the Lepidopteran midgut. To address the question of whether NICTABA is also active against Lepidopteran larvae, transgenic N. tabacum plants that silence endogenous NICTABA expression were constructed using RNA interference. Feeding experiments with these transgenic N. tabacum plants demonstrated that silencing of NICTABA expression enhances the larval performance of the generalist pest insect Spodoptera littoralis. In a second transgenic approach, NICTABA was ectopically expressed in the wild diploid tobacco Nicotiana attenuata, a species that lacks a functional NICTABA gene. When these transgenic N. attenuata plants were used in feeding experiments with S. littoralis larvae, a clear reduction in mass gain and significantly slower development were observed. In addition, feeding experiments with the Solanaceae specialist, Manduca sexta, provided further evidence that NICTABA exerts clear entomotoxic effects on Lepidopteran larvae. PMID- 20018899 TI - Overexpression of OsRAN2 in rice and Arabidopsis renders transgenic plants hypersensitive to salinity and osmotic stress. AB - Nucleo-cytoplasmic partitioning of regulatory proteins is increasingly being recognized as a major control mechanism for the regulation of signalling in plants. Ras-related nuclear protein (Ran) GTPase is required for regulating transport of proteins and RNA across the nuclear envelope and also has roles in mitotic spindle assembly and nuclear envelope (NE) assembly. However, thus far little is known of any Ran functions in the signalling pathways in plants in response to changing environmental stimuli. The OsRAN2 gene, which has high homology (77% at the amino acid level) with its human counterpart, was isolated here. Subcellular localization results showed that OsRan2 is mainly localized in the nucleus, with some in the cytoplasm. Transcription of OsRAN2 was reduced by salt, osmotic, and exogenous abscisic acid (ABA) treatments, as determined by real-time PCR. Overexpression of OsRAN2 in rice resulted in enhanced sensitivity to salinity, osmotic stress, and ABA. Seedlings of transgenic Arabidopsis thaliana plants overexpressing OsRAN2 were overly sensitive to salinity stress and exogenous ABA treatment. Furthermore, three ABA- or stress-responsive genes, AtNCED3, AtPLC1, and AtMYB2, encoding a key enzyme in ABA synthesis, a phospholipase C homologue, and a putative transcriptional factor, respectively, were shown to have differentially induced expression under salinity and ABA treatments in transgenic and wild-type Arabidopsis plants. OsRAN2 overexpression in tobacco epidermal leaf cells disturbed the nuclear import of a maize (Zea mays L.) leaf colour transcription factor (Lc). In addition, gene-silenced rice plants generated via RNA interference (RNAi) displayed pleiotropic developmental abnormalities and were male sterile. PMID- 20018901 TI - Distortion of trichome morphology by the hairless mutation of tomato affects leaf surface chemistry. AB - Trichomes are specialized epidermal structures that function as physical and chemical deterrents against arthropod herbivores. Aerial tissues of cultivated tomato (Solanum lycopersicum) are populated by several morphologically distinct trichome types, the most abundant of which is the type VI glandular trichome that produces various specialized metabolites. Here, the effect of the hairless (hl) mutation on trichome density and morphology, chemical composition, and resistance to a natural insect herbivore of tomato was investigated. The results show that the major effect of hl on pubescence results from structural distortion (bending and swelling) of all trichome types in aerial tissues. Leaf surface extracts and isolated type VI glands from hl plants contained wild-type levels of monoterpenes, glycoalkaloids, and acyl sugars, but were deficient in sesquiterpene and polyphenolic compounds implicated in anti-insect defence. No choice bioassays showed that hl plants are compromised in resistance to the specialist herbivore Manduca sexta. These results establish a link between the morphology and chemical composition of glandular trichomes in cultivated tomato, and show that hl-mediated changes in these leaf surface traits correlate with decreased resistance to insect herbivory. PMID- 20018902 TI - Nicotiana tabacum EIL2 directly regulates expression of at least one tobacco gene induced by sulphur starvation. AB - Sulphur deficiency severely affects plant growth and their agricultural productivity leading to diverse changes in development and metabolisms. Molecular mechanisms regulating gene expression under low sulphur conditions remain largely unknown. AtSLIM1, a member of the EIN3-like (EIL) family was reported to be a central transcriptional regulator of the plant sulphur response, however, no direct interaction of this protein with any sulphur-responsive promoters was demonstrated. The focus of this study was on the analysis of a promoter region of UP9C, a tobacco gene strongly induced by sulphur limitation. Cloning and subsequent examination of this promoter resulted in the identification of a 20-nt sequence (UPE-box), also present in the promoters of several Arabidopsis genes, including three out of four homologues of UP9C. The UPE-box, consisting of two parallel tebs sequences (TEIL binding site), proved to be necessary to bind the transcription factors belonging to the EIL family and of a 5-nt conserved sequence at the 3'-end. The yeast one-hybrid analysis resulted in the identification of one transcription factor (NtEIL2) capable of binding to the UPE box. The interactions of NtEIL2, and its homologue from Arabidopsis, AtSLIM1, with DNA were affected by mutations within the UPE-box. Transient expression assays in Nicotiana benthamiana have further shown that both factors, NtEIL2 and AtSLIM1, activate the UP9C promoter. Interestingly, activation by NtEIL2, but not by AtSLIM1, was dependent on the sulphur-deficiency of the plants. PMID- 20018903 TI - Plant cell division is specifically affected by nitrotyrosine. AB - Virtually all eukaryotic alpha-tubulins harbour a C-terminal tyrosine that can be reversibly removed and religated, catalysed by a specific tubulin-tyrosine carboxypeptidase (TTC) and a specific tubulin-tyrosine ligase (TTL), respectively. The biological function of this post-translational modification has remained enigmatic. 3-nitro-L-tyrosine (nitrotyrosine, NO(2)Tyr), can be incorporated into detyrosinated alpha-tubulin instead of tyrosine, producing irreversibly nitrotyrosinated alpha-tubulin. To gain insight into the possible function of detyrosination, the effect of NO(2)Tyr has been assessed in two plant model organisms (rice and tobacco). NO(2)Tyr causes a specific, sensitive, and dose-dependent inhibition of cell division that becomes detectable from 1 h after treatment and which is not observed with non-nitrosylated tyrosine. These effects are most pronounced in cycling tobacco BY-2 cells, where the inhibition of cell division is accompanied by a stimulation of cell length, and a misorientation of cross walls. NO(2)Tyr reduces the abundance of the detyrosinated form of alpha tubulin whereas the tyrosinated alpha-tubulin is not affected. These findings are discussed with respect to a model where NO(2)Tyr is accepted as substrate by TTL and subsequently blocks TTC activity. The irreversibly tyrosinated alpha-tubulin impairs microtubular functions that are relevant to cell division in general, and cell wall deposition in particular. PMID- 20018904 TI - Supply of sulphur to S-deficient young barley seedlings restores their capability to cope with iron shortage. AB - The effect of the S nutritional status on a plant's capability to cope with Fe shortage was studied in solution cultivation experiments in barley (Hordeum vulgare L. cv. Europa). Barley is a Strategy II plant and responds to Fe deficiency by secretion of chelating compounds, phytosiderophores (PS). All PS are derived from nicotianamine whose precursor is methionine. This suggests that a long-term supply of an inadequate amount of S could reduce a plant's capability to respond to Fe deficiency by limiting the rate of PS biosynthesis. The responses of barley (Hordeum vulgare L. cv. Europa) plants grown for 12 d on Fe free nutrient solutions (NS) containing 0 or 1.2 mM SO(4)(2-), was examined after 24 h or 48 h from transfer to NS containing 1.2 mM SO(4)(2-). After the supply of S was restored to S-deprived plants, an increase in PS release in root exudates was evident after 24 h of growth in S-sufficient NS and the increment reached values up to 4-fold higher than the control 48 h after S resupply. When S was supplied to S-deficient plants, leaf ATPS (EC 2.7.7.4) and OASTL (EC 4.2.99.8) activities exhibited a progressive recovery. Furthermore, root HvST1 transcript abundance remained high for 48 h following S resupply and a significant increase in the level of root HvYS1 transcripts was also found after only 24 h of S resupply. Data support the idea that the extent to which the plant is able to cope with Fe starvation is strongly associated with its S nutritional status. In particular, our results are indicative that barley plants fully recover their capability to cope with Fe shortage after the supply of S is restored to S deficient plants. PMID- 20018905 TI - Induced production of antifungal naphthoquinones in the pitchers of the carnivorous plant Nepenthes khasiana. AB - Nepenthes spp. are carnivorous plants that have developed insect capturing traps, evolved by specific modification of the leaf tips, and are able to utilize insect degradation products as nutritional precursors. A chitin-induced antifungal ability, based on the production and secretion to the trap liquid of droserone and 5-O-methyldroserone, is described here. Such specific secretion uniquely occurred when chitin injection was used as the eliciting agent and probably reflects a certain kind of defence mechanism that has been evolved for protecting the carnivory-based provision of nutritional precursors. The pitcher liquid containing droserone and 5-O-methyldroserone at 3:1 or 4:1 molar ratio, as well as the purified naphthoquinones, exerted an antifungal effect on a wide range of plant and human fungal pathogens. When tested against Candida and Aspergillus spp., the concentrations required for achieving inhibitory and fungicidal effects were significantly lower than those causing cytotoxicity in cells of the human embryonic kidney cell line, 293T. These naturally secreted 1,4-naphthoquinone derivatives, that are assumed to act via semiquinone enhancement of free radical production, may offer a new lead to develop alternative antifungal drugs with reduced selectable pressure for potentially evolved resistance. PMID- 20018906 TI - Late embryogenesis abundant (LEA) proteins in nondesiccated, encysted, and diapausing embryos of rotifers. AB - Two genes encoding for late embryogenesis abundant proteins (LEAs) are expressed in encysted diapausing embryos (or resting eggs) of rotifers (Brachionus plicatilis O.F. Muller) and females forming them. The two genes (bpa-leaa and bpa leab) share approximately 50% of their nucleotides sequence, and bpa-leaa is more than twofold longer than bpa-leab. The deduced amino acid sequences show high abundance of alanine, glycine, lysine, and glutamic acid; a hydropathy index of lower than one; and a relatively high (81-82%) predicted probability of forming alpha-helices in their secondary structure, all of which are characteristic features of LEAs. The predicted molecular masses of bpa-LEAA ( approximately 67 kDa) and bpa-LEAB ( approximately 27 kDa) are similar to the molecular mass determined by Western-blot analyses, suggesting a low probability of posttranslational modifications. In silico analysis reveals that the two LEAs resemble group 3 LEAs based on the repeats for 11mer motifs, although they also display several putative amino acids typical of the 20mer motif of group 1 LEAs. The rotifer LEAs do not contain a predicted target sequence and are more likely localized in the cytosol. LEAs were expressed in resting eggs and females producing them, but not in other female forms or males. LEA transcripts and proteins are degraded during hatching, suggesting that LEAs are developmentally programmed during resting egg formation and hatching. LEAs probably equip the resting eggs to withstand desiccation if that occurs during dormancy. The present study expands our knowledge about the biological pathways associated with formation of rotifer resting eggs and also demonstrates the occurrence of LEAs in dormant, nondesiccated, encysted animal embryos. PMID- 20018907 TI - Rescue of postcompaction-stage mouse embryo development from hypertonicity by amino acid transporter substrates that may function as organic osmolytes. AB - Early preimplantation embryos are sensitive to external osmolarity and use novel mechanisms to accumulate organic osmolytes and thus control their cell volumes and maintain viability. However, these mechanisms are restricted to the cleavage stages of development, and it was unknown whether postcompaction embryos use organic osmolytes. Mouse embryos developing from the 8-cell stage formed blastocoel cavities in vitro at osmolarities up to 360 mOsM. Above this range, several putative organic osmolytes (alanine, glutamine, glycine, and beta alanine) rescued blastocyst development, but several effective osmoprotectants in cleavage-stage embryos (such as betaine and proline) did not. At physiological osmolarities, each of these compounds resulted in significantly larger blastocysts. This was not due to increased cell numbers, which were unaffected in blastocysts by osmolarity in the range where blastocyst size was rescued by potential organic osmolytes, although cell number was decreased at higher osmolarities and was rescued by each osmolyte. The effective osmolytes were accumulated intracellularly by embryos developing in vitro from the 8-cell stage to blastocysts. However, unlike conventional organic osmolytes in somatic cells or those in cleavage-stage embryos, their intracellular concentrations were not increased with increasing external osmolarity. With the exception of beta alanine, which is taken up via the beta-amino acid transport system, the effective osmolytes were transported by the B(0,+) system, which becomes highly active in blastocysts. The intracellular accumulation of these osmolytes in postcompaction embryos thus appears to support optimal development and blastocyst expansion at physiological osmolarities and may contribute to the embryo's ability to withstand stress. PMID- 20018908 TI - Cloning of complementary DNAs encoding structurally related homeoproteins from preimplantation mouse embryos: their involvement in the differentiation of embryonic stem cells. AB - During the preimplantation development of mouse embryos between the 4-cell to 8 cell stage and the morula stage, when the first irreversible segregation of cell fates proceeds into the pluripotent inner cell mass (progenitor cells to form the fetus) and the trophectoderm (to form the placenta) of blastocysts, pluripotency maintaining and differentiation-inducing genes are expressed to coordinately regulate cell fates. Three structurally related cDNAs (Crxos1, Crxos1 sv2, and Crxos1 tv3) that exhibited concomitant elevated expression during this critical period were identified by subtractive cDNA cloning. CRXOS1 contains two homeodomains, while CRXOS1 sv2 and CRXOS1 tv3 each contain one of the homeodomains included in CRXOS1. Crxos1, Crxos1 sv2, and Crxos1 tv3 were expressed differentially during in vitro embryonic stem (ES) cell differentiation. Even under differentiation-inducing conditions, forced expression of Crxos1 sv2 inhibited the differentiation of ES cells. In contrast, under conditions that promote self-renewal of ES cells, forced expression of Crxos1 induced differentiation. Forced expression of Crxos1 resulted in induction of Gata4 but in repression of T, probably indicating that Crxos1 promotes the differentiation of ES cells into primitive endoderm, while inhibiting differentiation into mesoderm. On the other hand, no apparent effects of forced expression of Crxos1 tv3 were observed. Taken together, it was concluded that these transcripts encoding homeoproteins are capable of regulating the maintenance and/or differentiation of mouse ES cells and likely regulate that of preimplantation embryos. PMID- 20018909 TI - FOXP3+ regulatory T cells and T helper 1, T helper 2, and T helper 17 cells in human early pregnancy decidua. AB - In pregnancy, the decidua is infiltrated by leukocytes promoting fetal development without causing immunological rejection. Murine regulatory T (Treg) cells are known to be important immune regulators at this site. The aim of the study was to characterize the phenotype and origin of Treg cells and determine the quantitative relationship between Treg, T-helper type 1 (T(H)1), T(H)2, and T(H)17 cells in first-trimester human decidua. Blood and decidual CD4(+) T cells from 18 healthy first-trimester pregnant women were analyzed for expression of Treg-cell markers (CD25, FOXP3, CD127, CTLA4, and human leukocyte antigen-DR [HLA DR]), chemokine receptors (CCR4, CCR6, and CXCR3), and the proliferation antigen MKI67 by six-color flow cytometry. Treg cells were significantly enriched in decidua and displayed a more homogenous suppressive phenotype with more frequent expression of FOXP3, HLA-DR, and CTLA4 than in blood. More decidual Treg cells expressed MKI67, possibly explaining their enrichment at the fetal-maternal interface. Using chemokine receptor expression profiles of CCR4, CCR6, and CXCR3 as markers for T(H)1, T(H)2, and T(H)17 cells, we showed that T(H)17 cells were nearly absent in decidua, whereas T(H)2-cell frequencies were similar in blood and decidua. CCR6(+) T(H)1 cells, reported to secrete high levels of interferon gamma (IFNG), were fewer, whereas the moderately IFNG-secreting CCR6(-) T(H)1 cells were more frequent in decidua compared with blood. Our results point toward local expansion of Treg cells and low occurrence of T(H)17 cells. Furthermore, local, moderate T(H)1 activity seems to be a part of normal early pregnancy, consistent with a mild inflammatory environment controlled by Treg cells. PMID- 20018910 TI - ERBB receptor feedback inhibitor 1 regulation of estrogen receptor activity is critical for uterine implantation in mice. AB - Normal endometrial function requires a balance of progesterone (P4) and estrogen (E2) effects. E2 acts to stimulate the proliferation of uterine epithelial cells, while P4 action inhibits E2-mediated proliferation of the epithelium. P4 through its cognate receptor, the P4 receptor (Pgr), has important roles in the establishment and maintenance of pregnancy. In previous studies, we have identified ERBB receptor feedback inhibitor 1 (Errfi1) as a downstream target of Pgr action in the uterus. Herein, we show that Errfi1 mRNA expression was significantly increased in the uterus after Day 2.5 of gestation. Its expression is also induced in the uterus by acute E2 treatment, and this induction is synergistically induced by chronic E2 and P4 treatment. Although it is known that conditional ablation of Errfi1 in the Pgr-positive cells (Errfi1(d/d)) results in infertility, the function of Errfi1 in reproductive biology has remained elusive. Using Errfi1(d/d) mice, we have identified Errfi1 as an important mediator of uterine implantation. Epithelial ESR1 and target genes were significantly increased in the uteri of Errfi1(d/d) mice. Our results identify a new signaling paradigm of steroid hormone regulation in female reproductive biology that adds insight into the underlying dysregulation of hormonal signaling in human reproductive disorders such as endometriosis and endometrial cancer. PMID- 20018911 TI - Egg-citing advances in generating primordial germ cells in the laboratory. PMID- 20018912 TI - The protein C pathway in tissue inflammation and injury: pathogenic role and therapeutic implications. AB - Inflammation and coagulation are closely linked interdependent processes. Under physiologic conditions, the tissue microcirculation functions in anticoagulant and anti-inflammatory fashions. However, when inflammation occurs, coagulation is also set in motion and actively participates in enhancing inflammation. Recently, novel and unexpected roles of hemostasis in the humoral and cellular components of innate immunity have been described. In particular, the protein C system, besides its well-recognized role in anticoagulation, plays a crucial role in inflammation. Indeed, the protein C system is now emerging as a novel participant in the pathogenesis of acute and chronic inflammatory diseases, such as sepsis, asthma, inflammatory bowel disease, atherosclerosis, and lung and heart inflammation, and may emerge as unexpected therapeutic targets for intervention. PMID- 20018913 TI - Very long-term outcome of acute promyelocytic leukemia after treatment with all trans retinoic acid and chemotherapy: the European APL Group experience. AB - Acute promyelocytic leukemia (APL) is highly curable with the combination of all trans retinoic acid (ATRA) and anthracycline-based chemotherapy (CT), but very long-term results of this treatment, when CT should be added to ATRA and the role of maintenance treatment, remain uncertain. In our APL93 trial that included 576 newly diagnosed APL patients, with a median follow-up of 10 years, 10-year survival was 77%. Maintenance treatment significantly reduced 10-year cumulative incidence of relapses, from 43.2% to 33%, 23.4%, and 13.4% with no maintenance, maintenance using intermittent ATRA, continuous 6 mercaptopurine plus methotrexate, and both treatments, respectively (P < .001). Maintenance particularly benefited patients with white blood cell (WBC) count higher than 5 x 10(9)/L (5000/microL). Early addition of CT to ATRA significantly improved 10 year event-free survival (EFS), but without significant effect on overall survival (OS). The 10-year cumulative incidence of deaths in complete response (CR), resulting mainly from myelosuppression, was 5.7%, 15.4%, and 21.7% in patients younger than 55, 55 to 65, and older than 65 years, respectively, supporting the need for less myelosuppressive treatments, particularly for consolidation therapy. This study is registered at http://clinicaltrials.gov as NCT00599937. PMID- 20018914 TI - Circulating microvesicles in B-cell chronic lymphocytic leukemia can stimulate marrow stromal cells: implications for disease progression. AB - Microvesicles (MVs) released by malignant cancer cells constitute an important part of the tumor microenvironment. They can transfer various messages to target cells and may be critical to disease progression. Here, we demonstrate that MVs circulating in plasma of B-cell chronic lymphocytic leukemia (CLL) patients exhibit a phenotypic shift from predominantly platelet derived in early stage to leukemic B-cell derived at advanced stage. Furthermore, the total MV level in CLL was significantly greater compared with healthy subjects. To understand the functional implication, we examined whether MVs can interact and modulate CLL bone marrow stromal cells (BMSCs) known to provide a "homing and nurturing" environment for CLL B cells. We found that CLL-MV can activate the AKT/mammalian target of rapamycin/p70S6K/hypoxia-inducible factor-1alpha axis in CLL-BMSCs with production of vascular endothelial growth factor, a survival factor for CLL B cells. Moreover, MV-mediated AKT activation led to modulation of the beta-catenin pathway and increased expression of cyclin D1 and c-myc in BMSCs. We found MV delivered phospho-receptor tyrosine kinase Axl directly to the BMSCs in association with AKT activation. This study demonstrates the existence of separate MV phenotypes during leukemic disease progression and underscores the important role of MVs in activation of the tumor microenvironment. PMID- 20018915 TI - IL-6 and MYC collaborate in plasma cell tumor formation in mice. AB - Interleukin-6 (IL-6) plays a critical role in the natural history of human plasma cell neoplasms (PCNs), such as plasma cell myeloma and plasmacytoma (PCT). IL-6 is also at the center of neoplastic plasma cell transformation in BALB/c (C) mice carrying a transgene, H2-L(d)-IL6, that encodes human IL-6 under control of the major histocompatibility complex H2-L(d) promoter: strain C.H2-L(d)-IL6. These mice are prone to PCT, but tumor development is incomplete with long latencies ( approximately 40% PCT at 12 months of age). To generate a more robust mouse model of IL-6-dependent PCN, we intercrossed strain C.H2-L(d)-IL6 with strains C.iMyc(Emu) or C.iMyc(Calpha), 2 interrelated gene-insertion models of the chromosomal T(12;15) translocation causing deregulated expression of Myc in mouse PCT. Deregulation of MYC is also a prominent feature of human PCN. We found that double-transgenic C.H2-L(d)-IL6/iMyc(Emu) and C.H2-L(d)-IL6/iMyc(Calpha) mice develop PCT with full penetrance (100% tumor incidence) and short latencies (3-6 months). The mouse tumors mimic molecular hallmarks of their human tumor counterparts, including elevated IL-6/Stat3/Bcl-X(L) signaling. The newly developed mouse strains may provide a good preclinical research tool for the design and testing of new approaches to target IL-6 in treatment and prevention of human PCNs. PMID- 20018916 TI - A highly efficient short hairpin RNA potently down-regulates CCR5 expression in systemic lymphoid organs in the hu-BLT mouse model. AB - Inhibiting the expression of the HIV-1 coreceptor CCR5 holds great promise for controlling HIV-1 infection in patients. Here we report stable knockdown of human CCR5 by a short hairpin RNA (shRNA) in a humanized bone marrow/liver/thymus (BLT) mouse model. We delivered a potent shRNA against CCR5 into human fetal liver derived CD34(+) hematopoietic progenitor/stem cells (HPSCs) by lentiviral vector transduction. We transplanted vector-transduced HPSCs solidified with Matrigel and a thymus segment under the mouse kidney capsule. Vector-transduced autologous CD34(+) cells were subsequently injected in the irradiated mouse, intended to create systemic reconstitution. CCR5 expression was down-regulated in human T cells and monocytes/macrophages in systemic lymphoid tissues, including gut associated lymphoid tissue, the major site of HIV-1 replication. The shRNA mediated CCR5 knockdown had no apparent adverse effects on T-cell development as assessed by polyclonal T-cell receptor Vbeta family development and naive/memory T-cell differentiation. CCR5 knockdown in the secondary transplanted mice suggested the potential of long-term hematopoietic reconstitution by the shRNA transduced HPSCs. CCR5 tropic HIV-1 infection was effectively inhibited in mouse derived human splenocytes ex vivo. These results demonstrate that lentiviral vector delivery of shRNA into human HPSCs could stably down-regulate CCR5 in systemic lymphoid organs in vivo. PMID- 20018917 TI - A novel Rag2-/-gammac-/--xenograft model of human CLL. AB - Easily reproducible animal models that allow for study of the biology of chronic lymphocytic leukemia (CLL) and to test new therapeutic agents have been very difficult to establish. We have developed a novel transplantable xenograft murine model of CLL by engrafting the CLL cell line MEC1 into Rag2(-/-)gamma(c)(-/-) mice. These mice lack B, T, and natural killer (NK) cells, and, in contrast to nude mice that retain NK cells, appear to be optimal recipient for MEC1 cells, which were successfully transplanted through either subcutaneous or intravenous routes. The result is a novel in vivo model that has systemic involvement, develops very rapidly, allows the measurement of tumor burden, and has 100% engraftment efficiency. This model closely resembles aggressive human CLL and could be very useful for evaluating both the biologic basis of CLL growth and dissemination as well as the efficacy of new therapeutic agents. PMID- 20018918 TI - Fetal hemoglobin in sickle cell anemia: genome-wide association studies suggest a regulatory region in the 5' olfactory receptor gene cluster. AB - In a genome-wide association study of 848 blacks with sickle cell anemia, we identified single nucleotide polymorphisms (SNPs) associated with fetal hemoglobin concentration. The most significant SNPs in a discovery sample were tested in a replication set of 305 blacks with sickle cell anemia and in subjects with hemoglobin E or beta thalassemia trait from Thailand and Hong Kong. A novel region on chromosome 11 containing olfactory receptor genes OR51B5 and OR51B6 was identified by 6 SNPs (lowest P = 4.7E-08) and validated in the replication set. An additional olfactory receptor gene, OR51B2, was identified by a novel SNP set enrichment analysis. Genome-wide association studies also validated a previously identified SNP (rs766432) in BCL11A, a gene known to affect fetal hemoglobin levels (P = 2.6E-21) and in Thailand and Hong Kong subjects. Elements within the olfactory receptor gene cluster might play a regulatory role in gamma-globin gene expression. PMID- 20018920 TI - Why did this happen to me? Cognitive schema disruption and posttraumatic stress disorder in victims of sexual trauma. AB - Many trauma researchers have proposed cognitive schemas as a heuristic device to understand the elusive process of integrating traumatic events. We examined the schemas of a sample (N = 257) of female participants classified by exposure to sexual trauma, nonsexual trauma, and no trauma experience. Cognitive schema was assessed with the Traumatic Stress Institute Belief Scale. As hypothesized, six schema subscales were significantly related to trauma group membership: Self Intimacy, Self-Safety, Self-Trust, Other Intimacy, Other Safety, and Other Trust. Moreover, cognitive schema operated as a partial mediator in the relationship between sexual trauma and posttraumatic stress disorder (PTSD) symptomatology. Results are discussed in terms of their implications for current theories of trauma and etiology of PTSD, and treatment interventions. PMID- 20018919 TI - High-dose cyclophosphamide for severe aplastic anemia: long-term follow-up. AB - Severe aplastic anemia (SAA) is a life-threatening bone marrow failure disorder that can be treated with bone marrow transplantation, immunosuppressive therapy, and high-dose cyclophosphamide. Here, we report long-term follow-up on 67 SAA patients (44 treatment-naive and 23 refractory) treated with high-dose cyclophosphamide. At 10 years, the overall actuarial survival was 88%, the response rate was 71% with the majority being complete, and the actuarial event free survival was 58% in 44 treatment-naive SAA patients. Patients with refractory SAA fared less well after high-dose cyclophosphamide therapy; at 10 years, overall actuarial survival, response, and actuarial event-free survival rates were 62%, 48%, and 27%, respectively. High-dose cyclophosphamide is highly effective therapy for severe aplastic anemia. Large randomized controlled trials will be necessary to establish how results of high-dose cyclophosphamide compare with either bone marrow transplantation or standard immunosuppressive regimens, such as antithymocyte globulin and cyclosporine. PMID- 20018921 TI - Sex differences in intimate partner violence and the use of coercive control as a motivational factor for intimate partner violence. AB - Research argues that coercive control (CC) is a special case of intimate partner violence (IPV). The present study hypothesized that instead CC is the motivator for other types of IPV, with control of the victim as the goal. When CC fails, physical types of IPV are used. This hypothesized relationship was tested using a large matched sample of 762 divorcing couples participating in divorce mediation. Structural equation modeling was used to analyze the data with CC predicting two latent common factors of the overall level of victimization separately for men and women. Significant causal relationships between CC and the latent construct of victimization for both members of the couples were found. In addition, CC, psychological abuse, sexual assault/intimidation/coercion, threats of and severe physical violence were disproportionately reported as perpetrated by men against women whereas reports of physical abuse (e.g., pushing, shoving, scratching) were not. PMID- 20018922 TI - Complement in antibody therapy: friend or foe? PMID- 20018923 TI - ITP has elevated BAFF expression. PMID- 20018924 TI - A mischief of mice. PMID- 20018925 TI - Multidrug resistance-associated protein 4 (MRP4) gene polymorphisms and treatment response in adult acute lymphoblastic leukemia. PMID- 20018927 TI - Central nervous system prophylaxis in mantle cell lymphoma. PMID- 20018929 TI - Restoration of the human stem cell niche after stem cell transplantation. PMID- 20018930 TI - Vascular complications after splenectomy for hematologic disorders. PMID- 20018932 TI - Deficits underlying impaired visually triggered step adjustments in mildly affected stroke patients. AB - BACKGROUND: The ability to make step adjustments while walking is often impaired following a stroke, but the basic sensorimotor control deficits responsible have not been established. OBJECTIVE: To identify these deficits in Patients who have recovered from stroke leaving only mild lower limb impairment. METHODS: Ten stroke and 10 age-matched control patients stepped onto an illuminated rectangle. In 40% of trials it jumped 140 mm either medially or laterally when the stepping foot left the ground, thus provoking a mid-step adjustment. In a separate block, patients performed the same task but with the body supported by a frame to eliminate balance responses. RESULTS: Irrespective of support condition stroke patients produced short-latency foot trajectory adjustments compatible with a fast-acting, possibly subcortical, visuomotor process. However, the latency was slightly but significantly longer for the contralesional leg (148 ms) than the ipsilesional leg (141 ms) and longer than for controls (129 ms). Stroke patients' foot adjustments were executed slower and undershot the target more than controls. These deficits were most pronounced in the medial direction when the body was unsupported. The pattern of undershooting was the same for ipsilesional and contralesional legs. CONCLUSIONS: Mildly impaired stroke patients have deficits in initiating and executing visually triggered step adjustments but more profound difficulties with balance control during the adjustment, which caused them to suppress mid-step adjustments of foot placement in the medial direction where balance demands were greatest. Paradoxically, such suppression outside the laboratory may also threaten balance if it leads to unsafe foot placement or obstacle collision. PMID- 20018933 TI - Bioinformatic analysis of gene sets regulated by ligand-activated and dominant negative peroxisome proliferator-activated receptor gamma in mouse aorta. AB - OBJECTIVE: Drugs that activate peroxisome proliferator-activated receptor (PPAR) gamma improve glucose sensitivity and lower blood pressure, whereas dominant negative mutations in PPARgamma cause severe insulin resistance and hypertension. We hypothesize that these PPARgamma mutants regulate target genes opposite to those of ligand-mediated activation, and we tested this hypothesis on a genomewide scale. METHODS AND RESULTS: We integrated gene expression data in aorta specimens from mice treated with the PPARgamma ligand rosiglitazone with data from mice containing a globally expressed knockin of the PPARgamma P465L dominant-negative mutation. We also integrated our data with publicly available data sets containing the following: (1) gene expression profiles in many human tissues, (2) PPARgamma target genes in 3T3-L1 adipocytes, and (3) experimentally validated PPARgamma binding sites throughout the genome. Many classic PPARgamma target genes were induced by rosiglitazone and repressed by dominant-negative PPARgamma. A similar pattern was observed for about 90% of the gene sets regulated by both rosiglitazone and dominant-negative PPARgamma. Genes exhibiting this pattern of contrasting regulation were significantly enriched for nearby PPARgamma binding sites. CONCLUSIONS: These results provide convincing evidence that the PPARgamma P465L mutation causes transcriptional effects that are opposite to those mediated by PPARgamma ligand, thus validating mice carrying the mutation as a model of PPARgamma interference. PMID- 20018934 TI - Pathways by which reconstituted high-density lipoprotein mobilizes free cholesterol from whole body and from macrophages. AB - OBJECTIVE: Reconstituted high-density lipoprotein (rHDL) is of interest as a potential novel therapy for atherosclerosis because of its ability to promote free cholesterol (FC) mobilization after intravenous administration. We performed studies to identify the underlying molecular mechanisms by which rHDL promote FC mobilization from whole body in vivo and macrophages in vitro. METHODS AND RESULTS: Wild-type (WT), SR-BI knockout (KO), ABCA1 KO, and ABCG1 KO mice received either rHDL or phosphate-buffered saline intravenously. Blood was drawn before and at several time points after injection for apolipoprotein A-I, phosphatidylcholine, and FC measurement. In WT mice, serum FC peaked at 20 minutes and rapidly returned toward baseline levels by 24 hours. Unexpectedly, ABCA1 KO and ABCG1 KO mice did not differ from WT mice regarding the kinetics of FC mobilization. In contrast, in SR-BI KO mice the increase in FC level at 20 minutes was only 10% of that in control mice (P<0.01). Bone marrow-derived macrophages from WT, SR-BI O, ABCA1 KO, and ABCG1 KO mice were incubated in vitro with rHDL and cholesterol efflux was determined. Efflux from SR-BI KO and ABCA1 KO macrophages was not different from WT macrophages. In contrast, efflux from ABCG1 KO macrophages was approximately 50% lower as compared with WT macrophages (P<0.001). CONCLUSIONS: The bulk mobilization of FC observed in circulation after rHDL administration is primarily mediated by SR-BI. However, cholesterol mobilization from macrophages to rHDL is primarily mediated by ABCG1. PMID- 20018935 TI - Local atherosclerotic plaques are a source of prognostic biomarkers for adverse cardiovascular events. AB - OBJECTIVE: Atherosclerotic cardiovascular disease is a major burden to health care. Because atherosclerosis is considered a systemic disease, we hypothesized that one single atherosclerotic plaque contains ample molecular information that predicts future cardiovascular events in all vascular territories. METHODS AND RESULTS: AtheroExpress is a biobank collecting atherosclerotic lesions during surgery, with a 3-year follow-up. The composite primary outcome encompasses all cardiovascular events and interventions, eg, cardiovascular death, myocardial infarction, stroke, and endovascular interventions. A proteomics search identified osteopontin as a potential plaque biomarker. Patients undergoing carotid surgery (n=574) served as the cohort in which plaque osteopontin levels were examined in relation to their outcome during follow-up and was validated in a cohort of patients undergoing femoral endarterectomy (n=151). Comparing the highest quartile of carotid plaque osteopontin levels with quartile 1 showed a hazard ratio for the primary outcome of 3.8 (95% confidence interval, 2.6-5.9). The outcome did not change after adjustment for plaque characteristics and traditional risk factors (hazard ratio, 3.5; 95% confidence interval, 2.0-5.9). The femoral validation cohort showed a hazard ratio of 3.8 (95% confidence interval 2.0 to 7.4) comparing osteopontin levels in quartile 4 with quartile 1. CONCLUSIONS: Plaque osteopontin levels in single lesions are predictive for cardiovascular events in other vascular territories. Local atherosclerotic plaques are a source of prognostic biomarkers with a high predictive value for secondary manifestations of atherosclerotic disease. PMID- 20018936 TI - Phosphorylation and acetylation of histone H3 and autoregulation by early growth response 1 mediate interleukin 1beta induction of early growth response 1 transcription. AB - OBJECTIVE: The transcription factor early growth response (EGR)-1 has been implicated as a key vascular phenotypic switch through its control of inducible transcription. EGR-1 autoregulation, and histone modification in the EGR-1 promoter, represent key mechanisms in EGR-1 control, but have not been explored. METHODS AND RESULTS: We demonstrate that EGR-1 regulates its own transcription and that this involves histone H3 phosphorylation and acetylation. EGR-1 transactivates its promoter in smooth muscle cells exposed to interleukin (IL) 1beta through a novel cis-acting element (-211/-203). PD98059, which inhibits mitogen-activated protein kinase kinase/extracellular regulated kinase (MEK/ERK) attenuates IL-1beta-inducible phosphorylation of extracellular signal-regulated kinase 1/2 and mitogen and stress-activated protein kinases 1/2; and reduces levels of phosphorylated and acetylated histone H3. Histone deacetylase inhibition enhances EGR-1 transcription in response to cytokine. Conversely, suppression of histone modification with mitogen and stress-activated protein kinase 1/2 short interfering RNA, or the histone H3 acetyltransferase inhibitor Garcinol, inhibits IL-1beta-inducible EGR-1 transcription. EGR-1 interacts with the acetyltransferase p300. Acetylated H3 and phosphorylated H3 are enriched at the promoter of EGR-1; and EGR-1 is enriched at the promoters of tissue factor and plasminogen activator inhibitor 1 in response to IL-1beta, and attenuated by PD98059, Garcinol, and mitogen and stress-activated protein kinase 1/2 short interfering RNA. CONCLUSIONS: IL-1beta induction of EGR-1 transcription involves histone H3 phosphorylation, acetylation, and autoregulation by EGR-1. PMID- 20018938 TI - What is truth? Standards of scientific integrity in American Heart Association journals. PMID- 20018937 TI - Heat shock cognate protein 70 is essential for Akt signaling in endothelial function. AB - OBJECTIVE: Heat shock protein 70s (Hsp70s) are molecular chaperones that protect cells from damage in response to various stress stimuli. However, the functions and mechanisms in endothelial cells (ECs) have not been examined. Herein, we investigate the role of Hsp70s, including heat shock cognate protein 70 (Hsc70), which is constitutively expressed in nonstressed cells (ie, ECs). METHODS AND RESULTS: The Hsp70 inhibitor, KNK437, significantly decreased vascular endothelial growth factor (VEGF)-induced cell migration and tube formation in vitro. KNK437 inhibited the phosphorylation of VEGF-induced Akt and endothelial nitric oxide synthase (eNOS) in human umbilical vein endothelial cells. In a mouse hind limb model of vascular insufficiency, intramuscular inhibition of Hsp70s attenuated collateral and capillary vessel formation. Silencing the Hsc70 gene by short interfering RNA abolished VEGF-induced Akt phosphorylation and VEGF stimulated human umbilical vein endothelial cell migration and tube formation. As the molecular mechanisms, Hsc70 knockdown reduced the expression of phosphatidylinositol 3-kinase. CONCLUSIONS: Collectively, Hsc70 plays a significant role in ECs via the phosphatidylinositol 3-kinase/Akt pathway. Hsc70 may provide the basis for the development of new therapeutic strategies for angiogenesis. PMID- 20018939 TI - Lipolysis needed for chylomicron uptake? AB - Despite clinical evidence that postprandial lipemia and chylomicrons could contribute to atherosclerosis, direct evidence is lacking. The study by Weinstein et al(1) provides evidence to suggest that intact chylomicrons might be atherogenic by using genetically altered mice lacking Gpihbp1 protein, which may play a major role in the lipolysis of triglyceride-rich lipoproteins. However, the study does not rule out a contribution by remnants or limited lipolysis by other potential enzymes or pathways. It might be intriguing to determine the contribution of cholesterol derivatives in the chylomicrons to the lesions and the nature of the lesions. PMID- 20018940 TI - Smoking out the cause of thrombosis. PMID- 20018941 TI - Triglyceride-rich lipoproteins and plasma lipid transport. AB - This memoir provides a history of the triglyceride-rich lipoproteins of blood plasma over the last half-century. As precursors of low-density lipoproteins and in their own right, triglyceride-rich lipoproteins are essential to the formation of atherosclerotic plaques and to consequent ischemic vascular disease. The author recounts research at the National Heart Institute during 1953 to 1956 and continuing thereafter at the University of California San Francisco. Emphasis is placed on key insights arising from investigations of human disease, the interplay of fatty acid and triglyceride-transport involving the liver, small intestine, adipose tissue and muscle, and the role of the liver in the synthesis and catabolism of atherogenic lipoproteins. PMID- 20018942 TI - Glyco-conjugates as elicitors or suppressors of plant innate immunity. AB - Innate immunity is the first line of defense against invading microorganisms in vertebrates and the only line of defense in invertebrates and plants. Bacterial glyco-conjugates, such as lipopolysaccharides (LPS) from the outer membrane of Gram-negative bacteria and peptidoglycan (PGN) from the cell walls of both Gram positive and Gram-negative bacteria, and fungal and oomycete glycoconjugates such as oligosaccharides derived from the cell wall components beta-glucan, chitin and chitosan, have been found to act as elicitors of plant innate immunity. These conserved indispensable microbe-specific molecules are also referred to as microbe-associated molecular patterns (MAMPs). Other glyco-conjugates such as bacterial extracellular polysaccharides (EPS) and cyclic glucan have been shown to suppress innate immune responses, thus conversely promoting pathogenesis. MAMPs are recognized by the plant innate immune system though the action of pattern recognition receptors (PRRs). A greater insight into the mechanisms of MAMP recognition and the description of PRRs for different microbial glyco conjugates will have considerable impact on the improvement of plant health and disease resistance. Here we review the current knowledge about the bacterial MAMPs LPS and PGN, the fungal MAMPs beta-glucan, chitin and chitosan oligosaccharides and the bacterial suppressors EPS and cyclic glucan, with particular reference to the chemical structures of these molecules, the PRRs involved in their recognition (where these have been defined), and possible mechanisms underlying suppression. PMID- 20018943 TI - Characteristics of smoker support for increasing a dedicated tobacco tax: national survey data from New Zealand. AB - AIM: To examine smoker support for tobacco tax and for increased dedicated tobacco taxes, along with associations forany such support. METHODS: The New Zealand (NZ) arm of the International Tobacco Control Policy Evaluation Survey utilizes the NZ Health Survey (a national sample). From this sample, we surveyed adult smokers (N = 1,376). RESULTS: Most smokers considered that the current level of tobacco tax is "too high" (68%), but a majority (59%) would support an increase in tobacco tax if the extra revenue was used to promote healthy lifestyles and support quitting. There was majority support for a dedicated tobacco tax increase among all sociodemographic groups of smokers (including Maori, Pacific, and Asian smokers). In the fully adjusted multivariate model, significant associations with support for a dedicated tax increase included higher deprivation level (adjusted odds ratio [AOR] = 1.15) and suffering one form of financial stress (AOR = 1.81, 95% CI = 1.18-2.78). Other significant associations with support included concern about the smoking impacts on health and quality of life (AOR = 1.41), expressing support for tobacco control regulation (AOR = 1.83), and strength of intention to quit (AOR = 1.30). DISCUSSION: A majority of smokers from all sociodemographic groups supported an increase in tobacco tax if it was dedicated to quitting support and health promotion. The higher support among smokers with stronger intentions to quit is consistent with other evidence that smokers value tobacco control regulation such as high taxes to help them achieve their long-term quitting goals. PMID- 20018944 TI - The role of tobacco-specific media exposure, knowledge, and smoking status on selected attitudes toward tobacco control. AB - BACKGROUND: In August 2007, the President's Cancer Panel urged the leadership of the nation to "summon the political will to address the public health crisis caused by tobacco use" (President's Cancer Panel, N, 2007, Promoting healthy lifestyles: Policy, program, and personal recommendations for reducing cancer risk. http://deainfo.nci.nih.gov/advisory/pcp/pcp07rpt/pcp07rpt.pdf). While some research has examined predictors of public support for tobacco control measures, little research has examined modifiable factors that may influence public attitudes toward tobacco control. METHODS: We used the American Legacy Foundation's 2003 American Smoking and Health Survey 2 to examine the contribution of smoking status, knowledge of the negative effects of tobacco, and tobacco-specific media exposure (antitobacco messages, news coverage of tobacco issues, and protobacco advertising) on U.S. adults' attitudes toward tobacco control. In addition, we assessed whether smoking status moderates the relationship between tobacco-specific media exposure and policy attitudes. Weighted multivariable logistic regression models were employed. RESULTS: Results suggest that knowledge of the negative effects of tobacco and smoking status are associated with attitudes toward tobacco control and that exposure to tobacco specific information in the media plays a role only in some instances. We found no evidence of effect modification by smoking status on the impact of exposure to tobacco-specific media on attitudes toward tobacco control. DISCUSSION: Understanding the impact of readily modifiable factors that shape policy attitudes is essential if we are to target outreach and education in a way that is likely to sway public support for tobacco control. PMID- 20018945 TI - Disparities in tobacco cessation medication orders and fills among special populations. AB - BACKGROUND: There is considerable interest in measuring and eliminating health care disparities among various special populations, but there is limited understanding of their extent, causes, or potential remedies. To improve this for tobacco cessation, we measured differences in the frequency of receiving and filling cessation medication prescriptions by race, ethnicity, age, language preference, health insurance, and pregnancy. METHODS: The relevant variables for all patients of a Minnesota medical group aged 18 years and older with clinician visits were extracted from the electronic medical records of 1 large medical group from March 2006 to February 2007. This was combined with claims data from 1 insurance plan that covered most of these individuals. Order and fill rates for cessation medications were then adjusted for each of the other variables. RESULTS: There were 32,733 current users of tobacco, 18,047 of whom had both health insurance and pharmacy claims data available. After adjustment, 15.4% overall had received an order for cessation medications during this year, but only 78% had filled it. Groups receiving fewer orders than their comparison groups were aged 18-34 years or older than 65 years, men, pregnant women, Asians and Hispanics, and those with non-English-language preference, on Medicaid, or with fewer visits. The same groups were less likely to fill that prescription, except patients with non-English preference or Medicaid. DISCUSSION: There are disparities in both the receipt of cessation medication orders and the likelihood of filling them for some special populations. The causes are likely to be complex, but this information provides a starting point for learning to improve this problem. PMID- 20018946 TI - Feasibility of a tobacco cessation intervention for pregnant Alaska Native women. AB - BACKGROUND: Among Alaska Native women residing in the Yukon-Kuskokwim (Y-K) Delta region of Western Alaska, about 79% smoke cigarettes or use smokeless tobacco during pregnancy. Treatment methods developed and evaluated among Alaska Native pregnant tobacco users do not exist. This pilot study used a randomized two-group design to assess the feasibility and acceptability of a targeted cessation intervention for Alaska Native pregnant women. METHODS: Recruitment occurred over an 8-month period. Enrolled participants were randomly assigned to the control group (n = 18; brief face-to-face counseling at the first visit and written materials) or to the intervention group (n = 17) consisting of face-to-face counseling at the first visit, four telephone calls, a video highlighting personal stories, and a cessation guide. Interview-based assessments were conducted at baseline and follow-up during pregnancy (>or=60 days postrandomization). Feasibility was determined by the recruitment and retention rates. RESULTS: The participation rate was very low with only 12% of eligible women (35/293) enrolled. Among enrolled participants, the study retention rates were high in both the intervention (71%) and control (94%) groups. The biochemically confirmed abstinence rates at follow-up were 0% and 6% for the intervention and control groups, respectively. DISCUSSION: The low enrollment rate suggests that the program was not feasible or acceptable. Alternative approaches are needed to improve the reach and efficacy of cessation interventions for Alaska Native women. PMID- 20018947 TI - Cigarette smoking and associated health risks among students at five universities. AB - INTRODUCTION: While most college students and other young adults who smoke fall into the light and intermittent smoking (LITS) category, they remain at risk for tobacco dependence and other adverse health effects from their smoking. This study examines smoking patterns, tobacco dependence, and other health variables among students at five universities to better understand how to identify and address tobacco use and related risks in a college health clinic setting. METHODS: A health screening survey was completed by 2,091 college and graduate student volunteers seeking routine care at their university health centers or participating in a health class. Independent health variables were analyzed descriptively and in regression analyses with three levels of smoking (none, non daily, and daily) and tobacco dependence to determine predictors and associated risks. RESULTS: Nearly a quarter of students reported any current smoking, 41% of whom reported smoking less than 1 cigarette/day (cpd). Of the daily smokers, 80% smoked less than 10 cpd but 45% met criteria for tobacco dependence. Any smoking was associated with high-risk alcohol use, risky driving, relational abuse, depression, less exercise, and utilization of emergency and mental health services. In regression analyses, students who experienced depression had more than double the odds of being dependent smokers (odds ratio [OR] = 2.32), as did those who reported abuse (OR = 2.07) or sought mental health counseling (OR = 2.09). DISCUSSION: Student health providers should be alerted to the multiple risks and comorbidities that occur among all smokers, including LITS, and intervene concurrently to help prevent or mitigate adverse outcomes that result from these conditions and behaviors. PMID- 20018948 TI - Actin polymerization in differentiated vascular smooth muscle cells requires vasodilator-stimulated phosphoprotein. AB - Our group has previously shown that vasoconstrictors increase net actin polymerization in differentiated vascular smooth muscle cells (dVSMC) and that increased actin polymerization is linked to contractility of vascular tissue (Kim et al., Am J Physiol Cell Physiol 295: C768-778, 2008). However, the underlying mechanisms are largely unknown. Here, we evaluated the possible functions of the Ena/vasodilator-stimulated phosphoprotein (VASP) family of actin filament elongation factors in dVSMC. Inhibition of actin filament elongation by cytochalasin D decreases contractility without changing myosin light-chain phosphorylation levels, suggesting that actin filament elongation is necessary for dVSM contraction. VASP is the only Ena/VASP protein highly expressed in aorta tissues, and VASP knockdown decreased smooth muscle contractility. VASP partially colocalizes with alpha-actinin and vinculin in dVSMC. Profilin, known to associate with G actin and VASP, also colocalizes with alpha-actinin and vinculin, potentially identifying the dense bodies and the adhesion plaques as hot spots of actin polymerization. The EVH1 domain of Ena/VASP is known to target these proteins to their sites of action. Introduction of an expressed EVH1 domain as a dominant negative inhibits stimulus-induced increases in actin polymerization. VASP phosphorylation, known to inhibit actin polymerization, is decreased during phenylephrine stimulation in dVSMC. We also directly visualized, for the first time, rhodamine-labeled actin incorporation in dVSMC and identified hot spots of actin polymerization in the cell cortex that colocalize with VASP. These results indicate a role for VASP in actin filament assembly, specifically at the cell cortex, that modulates contractility in dVSMC. PMID- 20018949 TI - The Akt isoforms are present at distinct subcellular locations. AB - Akt is involved in the regulation of diverse cellular functions such as cell proliferation, energy metabolism, and apoptosis. Although three Akt isoforms are known, the function of each isoform is poorly understood. To gain a better understanding of each Akt isoform, we examined the subcellular localization and expression of each isoform in transformed and nontransformed cells. Akt1 was localized in the cytoplasm, which is in agreement with the currently accepted model that cytoplasmic Akt is translocated and activated at the inner leaflet of the plasma membrane. Interestingly, HEK-293 and HEK-293T cells contained Akt1 in the nucleus and cytoplasm, respectively, suggesting that SV40 T-antigen plays a crucial role in the cytoplasmic localization and activation of Akt1 in HEK-293T. Akt2 was colocalized with the mitochondria, while Akt3 was localized in both the nucleus and nuclear membrane. The subcellular localization of the Akt isoforms was not substantially altered in response to ionizing radiation or EGF. Furthermore, the ablation of one Akt isoform by small interfering RNA (siRNA) did not alter the subcellular location of the remaining isoforms, suggesting that the major function of one isoform is not compensated for by other isoforms. Together, our data support the notion that Akt2 and Akt3 are regulated at the mitochondrial and nuclear membranes, respectively. The mitochondrial localization of Akt2 raises the possibility that this isoform may be involved in both glucose-based energy metabolism and suppression of apoptosis, two Akt functions previously identified with anti-pan-Akt antibodies. PMID- 20018950 TI - Calcium signaling via two-pore channels: local or global, that is the question. AB - Recently, we identified, for the first time, two-pore channels (TPCs, TPCN for gene name) as a novel family of nicotinic acid adenine dinucleotide phosphate (NAADP)-gated, endolysosome-targeted calcium release channels. Significantly, three subtypes of TPCs have been characterized, TPC1-3, with each being targeted to discrete acidic calcium stores, namely lysosomes (TPC2) and endosomes (TPC1 and TPC3). That TPCs act as NAADP-gated calcium release channels is clear, given that NAADP binds to high- and low-affinity sites associated with TPC2 and thereby induces calcium release and homologous desensitization, as observed in the case of endogenous NAADP receptors. Moreover, NAADP-evoked calcium signals via TPC2 are ablated by short hairpin RNA knockdown of TPC2 and by depletion of acidic calcium stores with bafilomycin. Importantly, however, NAADP-evoked calcium signals were biphasic in nature, with an initial phase of calcium release from lysosomes via TPC2, being subsequently amplified by calcium-induced calcium release (CICR) from the endoplasmic reticulum (ER). In marked contrast, calcium release via endosome-targeted TPC1 induced only spatially restricted calcium signals that were not amplified by CICR from the ER. These findings provide new insights into the mechanisms that cells may utilize to "filter" calcium signals via junctional complexes to determine whether a given signal remains local or is converted into a propagating global signal. Essentially, endosomes and lysosomes represent vesicular calcium stores, quite unlike the ER network, and TPCs do not themselves support CICR or, therefore, propagating regenerative calcium waves. Thus "quantal" vesicular calcium release via TPCs must subsequently recruit inositol 1,4,5-trisphoshpate receptors and/or ryanodine receptors on the ER by CICR to evoke a propagating calcium wave. This may call for a revision of current views on the mechanisms of intracellular calcium signaling. The purpose of this review is, therefore, to provide an appropriate framework for future studies in this area. PMID- 20018951 TI - Regulation of vascular smooth muscle cell calcification by extracellular pyrophosphate homeostasis: synergistic modulation by cyclic AMP and hyperphosphatemia. AB - Vascular calcification is a multifaceted process involving gain of calcification inducers and loss of calcification inhibitors. One such inhibitor is inorganic pyrophosphate (PP(i)), and regulated generation and homeostasis of extracellular PP(i) is a critical determinant of soft-tissue mineralization. We recently described an autocrine mechanism of extracellular PP(i) generation in cultured rat aortic vascular smooth muscle cells (VSMC) that involves both ATP release coupled to the ectophosphodiesterase/pyrophosphatase ENPP1 and efflux of intracellular PP(i) mediated or regulated by the plasma membrane protein ANK. We now report that increased cAMP signaling and elevated extracellular inorganic phosphate (P(i)) act synergistically to induce calcification of these VSMC that is correlated with progressive reduction in ability to accumulate extracellular PP(i). Attenuated PP(i) accumulation was mediated in part by cAMP-dependent decrease in ANK expression coordinated with cAMP-dependent increase in expression of TNAP, the tissue nonselective alkaline phosphatase that degrades PP(i). Stimulation of cAMP signaling did not alter ATP release or ENPP1 expression, and the cAMP-induced changes in ANK and TNAP expression were not sufficient to induce calcification. Elevated extracellular P(i) alone elicited only minor calcification and no significant changes in ANK, TNAP, or ENPP1. In contrast, combined with a cAMP stimulus, elevated P(i) induced decreases in the ATP release pathway(s) that supports ENPP1 activity; this resulted in markedly reduced rates of PP(i) accumulation that facilitated robust calcification. Calcified VSMC were characterized by maintained expression of multiple SMC differentiation marker proteins including smooth muscle (SM) alpha-actin, SM22alpha, and calponin. Notably, addition of exogenous ATP (or PP(i) per se) rescued cAMP + phosphate treated VSMC cultures from progression to the calcified state. These observations support a model in which extracellular PP(i) generation mediated by both ANK- and ATP release-dependent mechanisms serves as a critical regulator of VSMC calcification. PMID- 20018953 TI - ATP consumption by sarcoplasmic reticulum Ca2+ pumps accounts for 50% of resting metabolic rate in mouse fast and slow twitch skeletal muscle. AB - In this study, we aimed to directly quantify the relative contribution of Ca(2+) cycling to resting metabolic rate in mouse fast-twitch (extensor digitorum longus, EDL) and slow-twitch (soleus) skeletal muscle. Resting oxygen consumption of isolated muscles (Vo(2), microl.g wet wt(-1).s(-1)) measured polarographically at 30 degrees C was approximately 25% higher in soleus (0.61 +/- .03) than in EDL (0.46 +/- .03). To quantify the specific contribution of Ca(2+) cycling to resting metabolic rate, cyclopiazonic acid (CPA), a highly specific inhibitor of sarco(endo)plasmic reticulum Ca(2+) ATPases (SERCAs), was added to the bath at different concentrations (1, 5, 10, and 15 microM). There was a concentration dependent effect of CPA on Vo(2), with increasing CPA concentrations up to 10 microM resulting in progressively greater reductions in muscle Vo(2). There were no differences between 10 and 15 microM CPA, indicating that 10 microM CPA induces maximal inhibition of SERCAs in isolated muscle preparations. Relative reduction in muscle Vo(2) in response to CPA was nearly identical in EDL (1 microM, 10.6 +/- 3.0%; 5 microM, 33.2 +/- 3.4%; 10 microM, 49.2 +/- 2.9%; 15 microM, 50.9 +/- 2.1%) and soleus (1 microM, 11.2 +/- 1.5%; 5 microM, 37.7 +/- 2.4%; 10 microM, 50.0 +/- 1.3%; 15 microM, 49.9 +/- 1.6%). The results indicate that ATP consumption by SERCAs is responsible for approximately 50% of resting metabolic rate in both mouse fast- and slow-twitch muscles at 30 degrees C. Thus SERCA pumps in skeletal muscle could represent an important control point for energy balance regulation and a potential target for metabolic alterations to oppose obesity. PMID- 20018954 TI - Oxygen-wasting effect of inotropy: is there a need for a new evaluation? An experimental large-animal study using dobutamine and levosimendan. AB - BACKGROUND: We addressed the hypothesis that the inotropic drugs dobutamine and levosimendan both induce surplus oxygen consumption (oxygen wasting) relative to their contractile effect in equipotent therapeutic doses, with levosimendan being energetically more efficient. METHODS AND RESULTS: Postischemically reduced left ventricular function (stunning) was created by repetitive left coronary occlusions in 22 pigs. This contractile dysfunction was reversed by infusion of either levosimendan (24 microg/kg loading and 0.04 microg x kg(-1) x min(-1) infusion) or an equipotent dose of dobutamine (1.25 microg x kg(-1) x min(-1)). Contractility and cardiac output were normalized by both drug regimens. The energy cost of drug-induced contractility enhancement was assessed by myocardial oxygen consumption related to the mechanical indexes tension-time index, pressure volume area, and total mechanical energy. ANCOVA did not reveal any increased oxygen cost of contractility for either drug in these doses. However, both dobutamine and levosimendan at supratherapeutic levels (10 microg x kg(-1) x min( 1) and 48 microg/kg loading with 0.2 microg x kg(-1) x min(-1) infusion, respectively) induced a highly significant increase in oxygen consumption related to mechanical work, compatible with the established oxygen-wasting effect of inotropy (P<0.001 for all mechanical indexes with dobutamine; P=0.007 for levosimendan as assessed by pressure-volume area). CONCLUSIONS: Therapeutic levels of neither dobutamine nor levosimendan showed inotropic oxygen wasting in this in vivo pig model. Thus, relevant hemodynamic responses can be achieved with an adrenergic inotrope without surplus oxygen consumption. PMID- 20018952 TI - Tetramerization domain mutations in KCNA5 affect channel kinetics and cause abnormal trafficking patterns. AB - The activity of voltage-gated K(+) (K(V)) channels plays an important role in regulating pulmonary artery smooth muscle cell (PASMC) contraction, proliferation, and apoptosis. The highly conserved NH(2)-terminal tetramerization domain (T1) of K(V) channels is important for proper channel assembly, association with regulatory K(V) beta-subunits, and localization of the channel to the plasma membrane. We recently reported two nonsynonymous mutations (G182R and E211D) in the KCNA5 gene of patients with idiopathic pulmonary arterial hypertension, which localize to the T1 domain of KCNA5. To study the electrophysiological properties and expression patterns of the mutants compared with the wild-type (WT) channel in vitro, we transfected HEK-293 cells with WT KCNA5, G182R, E211D, or the double mutant G182R/E211D channel. The mutants form functional channels; however, whole cell current kinetic differences between WT and mutant channels exist. Steady-state inactivation curves of the G182R and G182R/E211D channels reveal accelerated inactivation; the mutant channels inactivated at more hyperpolarized potentials compared with the WT channel. Channel protein expression was also decreased by the mutations. Compared with the WT channel, which was present in its mature glycosylated form, the mutant channels are present in greater proportion in their immature form in HEK-293 cells. Furthermore, G182R protein level is greatly reduced in COS-1 cells compared with WT. Immunostaining data support the hypothesis that, while WT protein localizes to the plasma membrane, mutant protein is mainly retained in intracellular packets. Overall, these data support a role for the T1 domain in channel kinetics as well as in KCNA5 channel subcellular localization. PMID- 20018957 TI - Rallying the caucus--the Democrats' struggle for unity on reform. PMID- 20018955 TI - Cardiac-specific overexpression of catalase identifies hydrogen peroxide dependent and -independent phases of myocardial remodeling and prevents the progression to overt heart failure in G(alpha)q-overexpressing transgenic mice. AB - BACKGROUND: Although it seems that reactive oxygen species contribute to chronic myocardial remodeling, questions remain about (1) the specific types of reactive oxygen species involved, (2) the role of reactive oxygen species in mediating specific cellular events, and (3) the cause-and-effect relationship between myocardial reactive oxygen species and the progression to heart failure. Transgenic mice with myocyte-specific overexpression of G(alpha)q develop a dilated cardiomyopathy that progresses to heart failure. We used this model to examine the role of H(2)O(2) in mediating myocardial remodeling and the progression to failure. METHODS AND RESULTS: In G(alpha)q myocardium, markers of oxidative stress were increased at 4 weeks and increased further at 20 weeks. G(alpha)q mice were crossbred with transgenic mice having myocyte-specific overexpression of catalase. At 4 weeks of age, left ventricular end-diastolic dimension was increased and left ventricular fractional shortening decreased in G(alpha)q mice and deteriorated further through 20 weeks. In G(alpha)q mice, myocardial catalase overexpression had no effect on left ventricular end diastolic dimension or fractional shortening at 4 weeks but prevented the subsequent deterioration in both. In G(alpha)q mice, myocyte hypertrophy; myocyte apoptosis; interstitial fibrosis; and the progression to overt heart failure, as reflected by lung congestion and exercise intolerance, were prevented by catalase overexpression. CONCLUSIONS: In G(alpha)q mice, myocyte-specific overexpression of catalase had no effect on the initial phenotype of left ventricular dilation and contractile dysfunction but prevented the subsequent progressive remodeling phase leading to heart failure. Catalase prevented the cellular hallmarks of adverse remodeling (myocyte hypertrophy, myocyte apoptosis, and interstitial fibrosis) and the progression to overt heart failure. Thus, H(2)O(2), associated oxidant pathways, or both play a critical role in adverse myocardial remodeling and the progression to failure. PMID- 20018958 TI - Lack of population diversity in commonly used human embryonic stem-cell lines. PMID- 20018960 TI - Ending end-of-life phobia--a prescription for enlightened health care reform. PMID- 20018959 TI - MMP12, lung function, and COPD in high-risk populations. AB - BACKGROUND: Genetic variants influencing lung function in children and adults may ultimately lead to the development of chronic obstructive pulmonary disease (COPD), particularly in high-risk groups. METHODS: We tested for an association between single-nucleotide polymorphisms (SNPs) in the gene encoding matrix metalloproteinase 12 (MMP12) and a measure of lung function (prebronchodilator forced expiratory volume in 1 second [FEV(1)]) in more than 8300 subjects in seven cohorts that included children and adults. Within the Normative Aging Study (NAS), a cohort of initially healthy adult men, we tested for an association between SNPs that were associated with FEV(1) and the time to the onset of COPD. We then examined the relationship between MMP12 SNPs and COPD in two cohorts of adults with COPD or at risk for COPD. RESULTS: The minor allele (G) of a functional variant in the promoter region of MMP12 (rs2276109 [-82A-->G]) was positively associated with FEV(1) in a combined analysis of children with asthma and adult former and current smokers in all cohorts (P=2x10(-6)). This allele was also associated with a reduced risk of the onset of COPD in the NAS cohort (hazard ratio, 0.65; 95% confidence interval [CI], 0.46 to 0.92; P=0.02) and with a reduced risk of COPD in a cohort of smokers (odds ratio, 0.63; 95% CI, 0.45 to 0.88; P=0.005) and among participants in a family-based study of early-onset COPD (P=0.006). CONCLUSIONS: The minor allele of a SNP in MMP12 (rs2276109) is associated with a positive effect on lung function in children with asthma and in adults who smoke. This allele is also associated with a reduced risk of COPD in adult smokers. PMID- 20018962 TI - Matrix metalloproteinase 12, asthma, and COPD. PMID- 20018963 TI - A common genetic fingerprint in leprosy and Crohn's disease? PMID- 20018961 TI - Genomewide association study of leprosy. AB - BACKGROUND: The narrow host range of Mycobacterium leprae and the fact that it is refractory to growth in culture has limited research on and the biologic understanding of leprosy. Host genetic factors are thought to influence susceptibility to infection as well as disease progression. METHODS: We performed a two-stage genomewide association study by genotyping 706 patients and 1225 controls using the Human610-Quad BeadChip (Illumina). We then tested three independent replication sets for an association between the presence of leprosy and 93 single-nucleotide polymorphisms (SNPs) that were most strongly associated with the disease in the genomewide association study. Together, these replication sets comprised 3254 patients and 5955 controls. We also carried out tests of heterogeneity of the associations (or lack thereof) between these 93 SNPs and disease, stratified according to clinical subtype (multibacillary vs. paucibacillary). RESULTS: We observed a significant association (P<1.00x10(-10)) between SNPs in the genes CCDC122, C13orf31, NOD2, TNFSF15, HLA-DR, and RIPK2 and a trend toward an association (P=5.10x10(-5)) with a SNP in LRRK2. The associations between the SNPs in C13orf31, LRRK2, NOD2, and RIPK2 and multibacillary leprosy were stronger than the associations between these SNPs and paucibacillary leprosy. CONCLUSIONS: Variants of genes in the NOD2-mediated signaling pathway (which regulates the innate immune response) are associated with susceptibility to infection with M. leprae. PMID- 20018964 TI - Uncomfortable arithmetic--whom to cover versus what to cover. PMID- 20018965 TI - Images in clinical medicine: Hernia through the foramen omentale. PMID- 20018967 TI - Images in clinical medicine: Frostbite. PMID- 20018966 TI - Molecular origins of cancer: Molecular basis of colorectal cancer. PMID- 20018968 TI - Case records of the Massachusetts General Hospital: Case 39-2009--a 28-year-old pregnant woman with acute cardiac failure. PMID- 20018969 TI - The agenda for continuing medical education--limiting industry's influence. PMID- 20018970 TI - Gene therapy for color blindness. PMID- 20018971 TI - Treatment of lung cancer. PMID- 20018972 TI - Asymptomatic reactivation of JC virus in patients treated with natalizumab. PMID- 20018973 TI - Coronary calcium screening. PMID- 20018974 TI - Proposals for payment reform in Massachusetts. PMID- 20018975 TI - Simulation model predicts cost savings from colorectal cancer screening. PMID- 20018976 TI - Mixed results on link between cellular telephones and cancer. PMID- 20018977 TI - Study finds omission of axillary lymph node dissection safe for patients with sentinel node micrometastases. PMID- 20018978 TI - Positions under positive selection--key for selectivity and potency of scorpion alpha-toxins. AB - Alpha-neurotoxins target voltage-gated sodium channels (Na(v)s) and constitute an important component in the venom of Buthidae scorpions. These toxins are short polypeptides highly conserved in sequence and three-dimensional structure, and yet they differ greatly in activity and preference for insect and various mammalian Na(v)s. Despite extensive studies of the structure-function relationship of these toxins, only little is known about their evolution and phylogeny. Using a broad data set based on published sequences and rigorous cloning, we reconstructed a reliable phylogenetic tree of scorpion alpha-toxins and estimated the evolutionary forces involved in the diversification of their genes using maximum likelihood-based methods. Although the toxins are largely conserved, four positions were found to evolve under positive selection, of which two (10 and 18; numbered according to LqhalphaIT and Lqh2 from the Israeli yellow scorpion Leiurus quinquestriatus hebraeus) have been previously shown to affect toxin activity. The putative role of the other two positions (39 and 41) was analyzed by mutagenesis of Lqh2 and LqhalphaIT. Whereas substitution P41K in Lqh2 did not alter its activity, substitution K41P in LqhalphaIT significantly decreased the activity at insect and mammalian Na(v)s. Surprisingly, not only that substitution A39L in both toxins increased their activity by 10-fold but also LqhalphaIT(A39L) was active at the mammalian brain channel rNa(v)1.2a, which otherwise is hardly affected by LqhalphaIT, and Lqh2(A39L) was active at the insect channel, DmNa(v)1, which is almost insensitive to Lqh2. Thus, position 39 is involved not only in activity but also in toxin selectivity. Overall, this study describes evolutionary forces involved in the diversification of scorpion alpha-toxins, highlights the key role of positions under positive selection for selectivity and potency, and raises new questions as to the toxin-channel face of interaction. PMID- 20018980 TI - Nineteenth century seeds reveal the population genetics of landrace barley (Hordeum vulgare). AB - Barley (Hordeum vulgare) is a major crop, grown worldwide and in a wide range of climatic conditions. Despite its importance as a crop species, little is known about the population genetics of barley and the effects of bottlenecks, adaptation, and gene flow on genetic diversity within and between landrace populations. In areas with highly developed agriculture, such as Northern Europe, these types of genetic studies are hampered by lack of landraces preserved in situ or ex situ. Here, we report a genetic study of Swedish landrace barley using 113-year-old seed samples. The results demonstrate differing levels of variation with some latitudinal effect. We also detect clear population differentiation and population structure within Sweden into a southern and a northern cluster. These results possibly reflect different introduction routes of barley into Sweden. We thus show that the study of historic material can be an important alternative for regions where no or little extant landrace material is available. PMID- 20018979 TI - Modal codon usage: assessing the typical codon usage of a genome. AB - Most genomes are heterogeneous in codon usage, so a codon usage study should start by defining the codon usage that is typical to the genome. Although this is commonly taken to be the genomewide average, we propose that the mode-the codon usage that matches the most genes-provides a more useful approximation of the typical codon usage of a genome. We provide a method for estimating the modal codon usage, which utilizes a continuous approximation to the number of matching genes and a simplex optimization. In a survey of bacterial and archaeal genomes, as many as 20% more of the genes in a given genome match the modal codon usage than the average codon usage. We use the mode to examine the evolution of the multireplicon genomes of Agrobacterium tumefaciens C58 and Borrelia burgdorferi B31. In A. tumefaciens, the circular and linear chromosomes are characterized by a common "chromosome-like" codon usage, whereas both plasmids share a distinct "plasmid-like" codon usage. In B. burgdorferi, in addition to different codon usage biases on the leading and lagging strands of DNA replication found by McInerney (McInerney JO. 1998. Replicational and transcriptional selection on codon usage in Borrelia burgdorferi. Proc Natl Acad Sci USA. 95:10698-10703), we also detect a codon-usage similarity between linear plasmid lp38 and the leading strand of the chromosome and a high similarity among the cp32 family of plasmids. PMID- 20018981 TI - Comparative testing of DNA segmentation algorithms using benchmark simulations. AB - Numerous segmentation methods for the detection of compositionally homogeneous domains within genomic sequences have been proposed. Unfortunately, these methods yield inconsistent results. Here, we present a benchmark consisting of two sets of simulated genomic sequences for testing the performances of segmentation algorithms. Sequences in the first set are composed of fixed-sized homogeneous domains, distinct in their between-domain guanine and cytosine (GC) content variability. The sequences in the second set are composed of a mosaic of many short domains and a few long ones, distinguished by sharp GC content boundaries between neighboring domains. We use these sets to test the performance of seven segmentation algorithms in the literature. Our results show that recursive segmentation algorithms based on the Jensen-Shannon divergence outperform all other algorithms. However, even these algorithms perform poorly in certain instances because of the arbitrary choice of a segmentation-stopping criterion. PMID- 20018983 TI - Why don't we practice what we preach? A meta-analytic review of religious racism. AB - A meta-analytic review of past research evaluated the link between religiosity and racism in the United States since the Civil Rights Act. Religious racism partly reflects intergroup dynamics. That is, a strong religious in-group identity was associated with derogation of racial out-groups. Other races might be treated as out-groups because religion is practiced largely within race, because training in a religious in-group identity promotes general ethnocentrism, and because different others appear to be in competition for resources. In addition, religious racism is tied to basic life values of social conformity and respect for tradition. In support, individuals who were religious for reasons of conformity and tradition expressed racism that declined in recent years with the decreased societal acceptance of overt racial discrimination. The authors failed to find that racial tolerance arises from humanitarian values, consistent with the idea that religious humanitarianism is largely expressed to in-group members. Only religious agnostics were racially tolerant. PMID- 20018982 TI - Novel acyl-coenzyme A:monoacylglycerol acyltransferase plays an important role in hepatic triacylglycerol secretion. AB - Acyl-CoA:monoacylglycerol acyltransferase (MGAT) plays a predominant role in the resynthesis of triacylglycerol in the small intestine, but its contribution to triacylglycerol synthesis in other tissues, such as the liver, is not clear. In this study, we identified a novel MGAT gene, which is identical with lysophosphatidylglycerol acyltransferase1 (LPGAT1). Mouse LPGAT1 is expressed in a number of tissues and most highly expressed in the liver. Hepatic LPGAT1 expression in diabetic db/db mice is higher than that in the control db/m mouse, which is consistent with increased hepatic MGAT activity in db/db mouse. To elucidate the role of LPGAT1 gene in lipid metabolism in db/db mice, we constructed an adenovirus of short hairpin RNA (shRNA) targeting LPGAT1 to selectively knockdown LPGAT1 gene expression in the liver. Hepatic MGAT activity and LPGAT1 expression in db/db mice infected with LPGAT1 shRNA adenovirus were significantly lower than those in mice infected with the control virus. Notably, treatment with LPGAT1 shRNA adenovirus caused a marked reduction in serum triacylglycerol and cholesterol levels and a significant increase in hepatic cholesterol level. These findings indicate that LPGAT1, a newly identified MGAT enzyme, plays a significant role in hepatic triacylglycerol synthesis and secretion in db/db mice. PMID- 20018984 TI - Age-related variation in carbon allocation at tree and stand scales in beech (Fagus sylvatica L.) and sessile oak (Quercus petraea (Matt.) Liebl.) using a chronosequence approach. AB - Two types of physiological mechanisms can contribute to growth decline with age: (i) the mechanisms leading to the reduction of carbon assimilation (input) and (ii) those leading to modification of the resource economy. Surprisingly, the processes relating to carbon allocation have been little investigated as compared to research on the processes governing carbon assimilation. The objective of this paper was thus to test the hypothesis that growth decrease related to age is accompanied by changes in carbon allocation to the benefit of storage and reproductive functions in two contrasting broad-leaved species: beech (Fagus sylvatica L.) and sessile oak (Quercus petraea (Matt.) Liebl.). Age-related changes in carbon allocation were studied using a chronosequence approach. Chronosequences, each consisting of several even-aged stands ranging from 14 to 175 years old for beech and from 30 to 134 years old for sessile oak, were divided into five or six age classes. In this study, carbon allocations to growth, storage and reproduction were defined as the relative amount of carbon invested in biomass increment, carbohydrate increment and seed production, respectively. Tree-ring width and allometric relationships were used to assess biomass increment at the tree and stand scales. Below-ground biomass was assessed using a specific allometric relationship between root:shoot ratio and age, established from the literature review. Seasonal variations of carbohydrate concentrations were used to assess carbon allocation to storage. Reproduction effort was quantified for beech stands by collecting seed and cupule production. Age-related flagging of biomass productivity was assessed at the tree and stand scales, and carbohydrate quantities in trees increased with age for both species. Seed and cupule production increased with stand age in beech from 56 gC m(-)(2) year(-1) at 30 years old to 129 gC m(-2) year(-1) at 138 years old. In beech, carbon allocation to storage and reproductive functions increased with age to the detriment of carbon allocation to growth functions. In contrast, the carbon balance between growth and storage remained constant between age classes in sessile oak. The contrasting age-related changes in carbon allocation between beech and sessile oak are discussed with reference to the differences in growing environment, phenology and hydraulic properties of ring-porous and diffuse-porous species. PMID- 20018985 TI - Federalism and technological change in blood products. AB - Recent research has shown how federalism affects health care finance, health care reform, and health policy innovation. The purpose of this article is to extend this research program to study the linkages between federalism and technological change. It does so using comparative case studies spanning five countries to examine innovation and diffusion of two blood technologies-enzyme-linked immunosorbent assays (ELISA blood tests) and heat treatment-in response to the threat to the blood supply posed by HIV during the 1980s. Prior research has produced three contradictory models of the federalism-innovation relationship. This article attempts to resolve these contradictions, posits new hypotheses, and highlights sources of omitted variable bias that have important implications for understanding technological change. The case studies show that overall decentralization, rather than federalism alone, aids technological progress by allowing its supporters to "venue shop" around political resistance. Decentralization also makes the state less vulnerable to capture by status-quo interest groups. Moreover, political decentralization may have a positive effect on technological diffusion, but a far weaker effect on innovation. Thus, prior research that conflates these two effects should be revisited. PMID- 20018986 TI - Maximizing federal Medicaid dollars: nursing home provider tax adoption, 2000 2004. AB - Since Medicaid is jointly financed by the federal and state governments, state officials have sought to offset state expenditures by maximizing federal contributions. One such strategy is to adopt a provider tax, which enables states to collect revenues from providers; those revenues are then used to pay for services rendered to Medicaid recipients, thereby leveraging federal matching dollars without concomitant increases in state expenditures. The number of states adopting a nursing home tax increased from thirteen to thirty-one between 2000 and 2004. This study seeks to identify the factors that spurred the rapid increase in nursing home provider taxes following implementation of the Balanced Budget Act of 1997. Results indicate that states with more powerful nursing home lobbies, lower proportions of private pay nursing home residents, worse fiscal health, weaker fiscal capacity, broader Medicaid eligibility, and nursing home supply restrictions were more likely to adopt. This implies that state officials react rationally to prevailing fiscal and programmatic circumstances when formulating policy under Medicaid and that providers seek relief, in part, from the adverse fiscal consequences of federal policy changes by promoting policy change at the state level. PMID- 20018987 TI - Partial progress: governing the pharmaceutical industry and the NHS, 1948-2008. AB - Coinciding with sixty years of the U.K. National Health Service (NHS), this article reviews the neglected area of the governance of the pharmaceutical industry and the NHS. It traces the relationships between the pharmaceutical industry, the state, and the NHS from the creation of the health service to the present, as they have grappled with the overlapping challenges of pharmaceutical safety, efficacy, cost-effectiveness, pricing, promotion, and advertising. The article draws on the concepts of "corporate bias" and "regulatory capture" from political theory, and "counter-vailing powers" and "clinical autonomy" in medical sociology, while also introducing the new concepts of "assimilated allies" and "pharmaceuticalization" in order to synthesize a theoretical framework capable of longitudinal empirical analysis of pharmaceutical governance. The analysis identifies areas in which the governance of pharmaceuticals and the NHS has contributed to progress in health care since 1948. However, it is argued that that progress has been slow, restricted, and vulnerable to misdirection due to the enormous and unrivaled influence afforded to the pharmaceutical industry in policy developments. Countervailing influences against such corporate bias have often been limited and subject to destabilization by the industry's assimilated allies either within the state or in the embrace of pharmaceuticalization and consumerism. PMID- 20018988 TI - Policy making on data exclusivity in the European Union: from industrial interests to legal realities. AB - After lengthening the duration of patents to twenty years in 1984, the pharmaceutical industry has turned to data exclusivity as a major vehicle for extending market protection, even after patents expire. Such protections give companies the power to tax consumers for innovation by charging above-market prices. This article draws upon unique information to describe how key actors lengthened data exclusivity for patented drugs to postpone generic competition in the European Union (EU) just before ten new members joined it. We explore the political route and the interests of different actors to understand the process by which industrial interests are translated into legal realities in the world's largest harmonized market. Several factors influenced the outcome, including the role of the pharmaceutical unit of the Directorate General for Enterprise of the European Commission in promoting the interests of the innovative branch of the industry, the time pressure to find a viable compromise before EU enlargement, and the heterogeneous preferences of the other actors. The case illustrates the inherent tension between the desire of both health care administrators and patients for high-quality, low-cost medicines and the objective of the innovator pharmaceutical industry to find and approve new drugs that are price protected and sell them in a way that maximizes revenues. PMID- 20018989 TI - Information and communications technology in U.S. health care: why is adoption so slow and is slower better? AB - Politicians across the political spectrum support greater investment in health care information and communications technology (ICT) and expect it to significantly decrease costs and improve health outcomes. We address three policy questions about adoption of ICT in health care: First, why is there so little adoption? Second, what policies will facilitate and accelerate adoption? Third, what is the best pace for adoption? We first describe the unusual economics of ICT, particularly network externalities, and then determine how those economics interact with and are exacerbated by the unusual economics of health care. High replacement costs and the need for technical compatibility are general barriers to ICT adoption and often result in lock-in to adopted technologies. These effects are compounded in health care because the markets for health care services, health insurance, and labor are interlinked. In addition, the government interacts with all markets in its role as an insurer. Patient heterogeneity further exacerbates these effects. Finally, ICT markets are often characterized by natural monopolies, resulting in little product diversity, an effect ill-suited to patient heterogeneity. The ongoing process for setting technical standards for health care ICT is critical but needs to include all relevant stakeholders, including patient groups. The process must be careful (i.e., slow), flexible, and allow for as much diversity as possible. We find that waiting to adopt ICT is a surprisingly wise policy. PMID- 20018990 TI - Performance-based regulation: enterprise responsibility for reducing death, injury, and disease caused by consumer products. AB - This article offers a bold new idea for confronting the staggering level of death, injury, and disease caused by five consumer products: cigarettes, alcohol, guns, junk food, and motor vehicles. Business leaders try to frame these negative outcomes as "collateral damage" that is someone else's problem. That framing not only is morally objectionable but also overlooks the possibility that, with proper prodding, industry could substantially lessen these public health disasters. I seek to reframe the public perception of who is responsible and propose to deploy a promising approach called "performance-based regulation" to combat the problem. Performance-based regulation would impose on manufacturers a legal obligation to reduce the negative social costs of their products. Rather than involving them in litigation or forcing them to operate differently (as "command-and-control" regimes do), performance-based regulation allows the firms to determine how best to decrease bad public health consequences. Like other public health strategies, performance-based regulation focuses on those who are far more likely than individual consumers to achieve real gains. Analogous to a tax on causing harm that exceeds a threshold level, performance-based regulation seeks to harness private initiative in pursuit of the public good. PMID- 20018992 TI - The confluence of evidence-based practice and Daubert within the fields of forensic psychiatry and the law. PMID- 20018993 TI - The justice and therapeutic promise of science-based research on criminal evil. PMID- 20018994 TI - Evaluating competency to stand trial with evidence-based practice. AB - Evaluations for competency to stand trial are distinguished from other areas of forensic consultation by their long history of standardized assessment beginning in the 1970s. As part of a special issue of the Journal on evidence-based forensic practice, this article examines three published competency measures: the MacArthur Competence Assessment Tool-Criminal Adjudication (MacCAT-CA), the Evaluation of Competency to Stand Trial-Revised (ECST-R), and the Competence Assessment for Standing Trial for Defendants with Mental Retardation (CAST-MR). Using the Daubert guidelines as a framework, we examined each competency measure regarding its relevance to the Dusky standard and its error and classification rates. The article acknowledges the past polarization of forensic practitioners on acceptance versus rejection of competency measures. It argues that no valuable information, be it clinical acumen or standardized data, should be systematically ignored. Consistent with the American Academy of Psychiatry and the Law Practice Guideline, it recommends the integration of competency interview findings with other sources of data in rendering evidence-based competency determinations. PMID- 20018995 TI - The criminal justice outcomes of jail diversion programs for persons with mental illness: a review of the evidence. AB - Diversion programs are initiatives in which persons with serious mental illness who are involved with the criminal justice system are redirected from traditional criminal justice pathways to the mental health and substance abuse treatment systems. This article is a review of the research literature conducted to determine whether the current evidence supports the use of diversion initiatives to reduce recidivism and to reduce incarceration among adults with serious mental illness with justice involvement. A structured literature search identified 21 publications or research papers for review that examined the criminal justice outcomes of various diversion models. The review revealed little evidence of the effectiveness of jail diversion in reducing recidivism among persons with serious mental illness. However, evidence was found that jail diversion initiatives can reduce the amount of jail time that persons with mental illness serve. Implications for practice and research are discussed. PMID- 20018996 TI - A meta-analysis of the psychological treatment of anger: developing guidelines for evidence-based practice. AB - There is no clear evidence to guide mental health professionals in assessing and treating angry clients. Recent reviews have considered cognitive and behavioral approaches to the treatment of anger, but little is known about the potential effectiveness of other treatment modalities. A meta-analytic review was conducted to examine the effects of treating dimensions of anger by using various psychological treatments found in the scientific literature. The final analysis included 96 studies and 139 treatment effects. The nine types of psychological treatments included cognitive, cognitive behavior therapy, exposure, psychodynamic, psychoeducational, relaxation-based, skills-based, stress inoculation, and multicomponent. The overall weighted standardized mean difference across all treatments was 0.76 (95% confidence interval [CI], 0.67 0.85, Q = 403.13, df 138, p < .001, I(2) = 65.76), which suggests that psychological treatments are generally effective in treating anger. The results also suggest a considerable degree of variability in the effect sizes of specific treatments for anger. The results show that at least some of the variability may be explained by the number of treatment sessions offered to participants, the use of manuals to guide delivery of the treatment, the use of fidelity checks, the setting of the research, and whether the study was published or unpublished. This review builds on previous evidence of the effectiveness of psychological treatments of maladaptive anger, and it provides the basis for developing evidence-based guidelines for specific populations with anger problems. PMID- 20018997 TI - Forensic psychiatry, neuroscience, and the law. AB - The rise of modern neuroscience is transforming psychiatry and other behavioral sciences. Neuroscientific progress also has had major impact in forensic neuropsychiatric practice, resulting in the increased use of neuroscientific technologies in cases of a psychiatric-legal nature. This article is focused on the impact of neuroscientific progress in forensic psychiatry in relation to criminal law. Also addressed are some emerging questions involving the practice of forensic neuropsychiatry. These questions will be reframed by providing alternative perspectives consistent with the objectives of forensic neuropsychiatric practice. The last part of the article is a discussion of potential developments that may facilitate the integration of neuroscientific knowledge in forensic neuropsychiatric practice. PMID- 20018998 TI - Commentary: evidence-based practice and forensic psychiatry. AB - A diverse sampling of articles was considered as a landscape against which evidence-based practice has been and should be a part of forensic psychiatry. Caveats were identified, limitations suggested, and recommendations made as to how such a marriage might work. PMID- 20018999 TI - A practical guide for the evaluation of sexual recidivism risk in mentally retarded sex offenders. AB - Although sex offender risk assessment has progressed greatly over the past decade and a half since most states implemented the sexually violent predator/sexually dangerous person (SVP/SDP) laws, there continues to be limited applicability of such models to intellectually disabled sex offenders because there has been no empirical validation. However, SVP/SDP civil commitment programs have reported increased admission of developmentally disabled sex offenders. Differentiating sexual deviance, the primary factor predisposing most individuals to criminal sexual violence, from impulsive, immature, and inappropriate behavior stemming from cognitive deficits presents yet another challenge to the clinician tasked with performing such evaluations. This article reviews actuarial risk models and their limited applicability to mentally retarded sex offenders and offers a conceptual method of assessing the risk of recidivism in intellectually disabled sex offenders under SVP/SDP evaluation. PMID- 20019000 TI - Credibility in the courtroom: how likeable should an expert witness be? AB - This study was conducted to investigate the relationship between expert witness likeability and jurors' judgments of credibility and tendencies in sentencing. Two actors playing expert witnesses were trained to present themselves as high and low in likeability in a standard testimony scenario in the sentencing phase of a capital murder trial. The effects of extraversion and gender of the 210 psychology undergraduates serving as mock jurors attending to expert testimony were also examined. The dependent variables were the jurors' perceptions of the witnesses' credibility and their agreement with the testimony. The likeability of the expert witnesses was found to be significantly related to the jurors' perception of their trustworthiness, but not to their displays of confidence or knowledge or to the mock jurors' sentencing decisions. The women rated highly likeable experts as more credible than the less likeable ones, but the men did not differentiate between the two types. As extraversion increased, the male jurors' agreement with testimony increased, but the female jurors' agreement decreased. The results suggest that likeability can be an important element in the credibility of the source and that attorneys and trial consultants now have an empirical foundation for addressing likeability as part of witness preparation. PMID- 20019001 TI - Allowing independent forensic evaluations for Guantanamo detainees. AB - President Obama has pledged to review all cases prosecuted through the Guantanamo Military Commissions established under President George N. Bush. Such commissions, however, may have limited independent psychiatric evaluations for Guantanamo defendants. This article explores the legal foundations for evaluating Guantanamo detainees, analyzes the decisions of commissions through discussions with defense attorneys involved in the cases, and considers the medicolegal consequences of the Guantanamo commissions. Recommendations are offered to safeguard the ethical soundness of future forensic consultations. PMID- 20019002 TI - White-collar crime: corporate and securities and commodities fraud. AB - In this era of increased interest in white-collar crime, forensic psychiatrists are in a key position to study the individual characteristics of offenders. While a comprehensive theory of high-level white-collar crime should consider societal and organizational contributions, there is value in understanding the personal traits that place an individual at high risk for offending. As the impact of the criminal acts of this group has been increasingly felt by larger groups from all socioeconomic strata, there is less willingness by the public to view these crimes as victimless and harmless. PMID- 20019003 TI - Gun laws and the involuntarily committed: a California road map. AB - The 2007 incident at Virginia Tech brought the question of gun ownership by the mentally ill to the forefront of public attention. Moreover, it underscored the potentially devastating consequences of the imperfect connection between federal and state laws that apply to the right of gun ownership by a psychiatric patient. The laws are complex, and, as demonstrated in this article, conflicting. We present a case report of an involuntarily committed patient in the state of California, and discuss details of state and federal laws that applied to him. PMID- 20019004 TI - Psychiatric and clinical sequelaeof delirium and competenceto stand trial. AB - We present the case of a middle-aged defendant who had been incarcerated in a county jail and housed on the mental health unit. It was documented that he had been exhibiting fluctuating levels of alertness and responsiveness. The writers saw him in a forensic capacity, to conduct an evaluation of his competence to stand trial, and recognized that he was having a medical emergency, delirium that was most likely due to brain metastases from inoperable advanced cancer. We recommended an immediate transfer to a medical facility for treatment. The article serves to present an interesting case and to highlight the need for clinical vigilance despite the usual goal of an objective, noninterfering forensic role. PMID- 20019005 TI - Forensic psychiatry and the forensic sciences: in memory of Peter J. Batten, MD. AB - This commentary is dedicated to the memory of Peter J. Batten, MD. He was a public health physician, a psychiatrist, and a medical examiner who spent his entire career in Salem, the capital of the State of Oregon. Salem was a unique place to work because, early in the history of Oregon, the state elected to build all of its original public institutions in the environs of the city. As the county medical examiner, Dr. Batten reviewed all questionable deaths that occurred within the county and in particular within the public institutions. Many of his findings were subsequently published, and these reports influenced the direction of mental health policy in these same institutions. He also used his position as county medical examiner to examine deaths in road rage incidents and those occurring at railroad crossings in Salem. The commentary also emphasizes the benefits of collaboration within the forensic sciences. PMID- 20019006 TI - Pathogenicity of Alternanthera mosaic virus is affected by determinants in RNA dependent RNA polymerase and by reduced efficacy of silencing suppression in a movement-competent TGB1. AB - Four biologically active cDNA clones were derived from the Alternanthera mosaic virus (AltMV; genus Potexvirus) isolate, AltMV-SP, which differ in symptoms in infected Nicotiana benthamiana plants. Two clones induced necrosis and plant death; a mixture of all four clones induced milder symptoms than AltMV-SP. Replication of all clones was enhanced by a minimum of fourfold at 15 degrees C. A mixture of clones 4-7 (severe) and 3-1 (mild) was indistinguishable from AltMV SP, but the ratio of 4-7 to 3-1 differed at 25 and 15 degrees C. RNA copy numbers of mixed infections were always below those of 4-7 alone. Determinants of symptom severity were identified in both Pol and TGB1; the mildest (4-1) and most severe (3-7) clones differed at three residues in the 'core' Pol domain [R(1110)P, K(1121)R, R(1255)K] and one [S(1535)P] in the C-terminal Pol domain of RNA dependent RNA polymerase, and one in TGB1 [P(88)L]. Pol [P(1110),R(1121),K(1255)]+TGB1(L(88))] always induced systemic necrosis at 15 degrees C. Gene exchanges of Pol and TGB1 each affected replication and symptom expression, with TGB1(P(88)) significantly reducing silencing suppression. The difference in silencing suppression between TGB1(P(88)) and TGB1(L(88)) was confirmed by an agroinfiltration assay. Further, co-expression of TGB1(P(88)) and TGB1(L(88)) resulted in interference in the suppression of silencing by TGB1(L(88)). Yeast two-hybrid analysis confirmed that TGB1(P(88)) and TGB1(L(88)) interact. These results identify a TGB1 residue that significantly affects replication and silencing suppression, but maintains full movement functions. PMID- 20019007 TI - Update on the treatment of saphenous reflux: laser, RFA or foam? AB - Chronic venous insufficiency resulting from great saphenous vein (GSV) reflux is a problem seen with increasing frequency in most vascular surgical practices. Traditionally treated with high ligation and stripping, minimally invasive techniques for ablation of the GSV have evolved. These include endovenous laser surgery, radiofrequency ablation, and foam sclerotherapy. The authors discuss the mechanism of action of each modality, as well as the techniques for intervention, outcomes, and complications. Minimally invasive treatment of venous insufficiency is efficacious, with less morbidity than high ligation and stripping, allowing for successful treatment and improved patient satisfaction. PMID- 20019008 TI - On the horizon: what's coming next for lower extremity interventions. AB - Multiple endovascular interventions exist for the treatment of lower extremity peripheral arterial disease (PAD), with some exhibiting better outcomes than others. As these procedures become more numerous and accepted, the type of interventionalist providing the care is changing. Despite advances in technology and techniques, there are many obstacles to overcome in order to improve the treatment of patients with vascular disease. These can be divided into managing recurrent disease, protection of the distal vasculature, and the prevention of disease. Studies are already underway in areas involving drug-eluting balloons and stents for the periphery, brachytherapy, bioabsorbable stents, distal protection, and therapeutic angiogenesis. As knowledge about the treatment of peripheral arterial disease is acquired that can be applied to the concept of personalized health care, there is hope to be able to slow or arrest the progression of this disease. PMID- 20019009 TI - E-only Erratum. PMID- 20019011 TI - Obligatory external quality control: a mandatory demand in device therapy. PMID- 20019012 TI - Successful catheter ablation of epicardial ventricular tachycardia worsened by cardiac resynchronization therapy. AB - Soon after an upgrade from a single-chamber implantable cardioverter defibrillator (ICD) to cardiac resynchronization therapy (CRT) with an ICD, a 64 year-old man with non-ischaemic cardiomyopathy began to have increasingly frequent ICD shocks for slow ventricular tachycardia (VT). At electrophysiological study, no clinical VT was induced by endocardial right ventricular pacing, but was easily induced by epicardial left ventricular (LV) pacing via a subxiphoid pericardial approach. The VT was successfully ablated on the LV epicardial surface. This case suggests that epicardial catheter ablation may be an alternative for managing CRT-induced proarrhythmias without the inactivation of LV pacing. PMID- 20019013 TI - Heart rhythm at the time of death documented by an implantable loop recorder. AB - AIMS: The aims of this study were to describe arrhythmias documented with an implantable loop recorder (ILR) in post-acute myocardial infarction (AMI) patients with left ventricular dysfunction at the time of death and to establish the correlation to mode of death. METHODS AND RESULTS: Post-mortem ILR device interrogations were analysed from patients dying in the CARISMA study. Mode of death was classified by a modified CAST classification. Twenty-six patients died with an implanted ILR. Of these, 16 had an electrocardiogram recorded at the time of death. Ventricular tachycardia (VT)/ventricular fibrillation (VF) was terminal rhythm in eight patients and bradyarrhythmias were observed in another eight patients. Of the deaths with peri-mortem recordings, seven were classified as sudden cardiac death (SCD). In six of these, VF was documented at the time of death. Six monitored deaths were classified as non-SCD (NSCD) of which only two had recordings of VT/VF, whereas four had bradyarrhythmias. All peri-mortem recordings in non-cardiac death (NCD) were bradyarrhythmia. CONCLUSION: Long-term monitoring in a population of post-AMI patients with left ventricular ejection fraction < or =40% showed that VT/VF and bradyarrhythmia each accounted for half of the recorded events at the time of death. The ILR confirmed that ventricular tachyarrhythmias are associated primarily with SCD, whereas bradyarrhythmias and electromechanical dissociation seems dominant in NSCD and NCD. The study was registered at ClinicalTrials.gov: NCT00145119. PMID- 20019014 TI - Mitral isthmus conduction block: intriguing result of radiofrequency catheter ablation for a left concealed accessory pathway. AB - We report on our experience with complete mitral isthmus conduction block achieved inadvertently during radiofrequency (RF) catheter ablation for a left lateral concealed accessory pathway (AP) mimicking concentric retrograde activation. This rare condition should be acknowledged to avoid misdiagnosis of another concomitant AP and to avoid RF applications in inappropriate areas. PMID- 20019015 TI - Use of a circular mapping and ablation catheter for ablation of atypical right ventricular outflow tract arrhythmia. AB - A new technique for ablation of persistent ectopic activity with atypical electrocardiographic characteristics at the vicinity of the right ventricular outflow tract is described. A new circular mapping and ablation catheter initially designed for pulmonary vein ablation was used. Abolition of ectopic activity was achieved with minimal fluoroscopy and ablation times. PMID- 20019016 TI - Methylamine clearance by haemodialysis is low. AB - BACKGROUND: Dialysis adequacy is currently judged by measures of urea clearance. However, urea is relatively non-toxic and has properties distinct from large classes of other retained solutes. In particular, intracellularly sequestered solutes are likely to behave differently than urea. METHODS: We studied an example of this class, the aliphatic amine monomethylamine (MMA), in stable haemodialysis outpatients (n = 10) using an HPLC-based assay. RESULTS: Mean MMA levels pre-dialysis in end-stage renal disease subjects were 76 +/- 15 microg/L compared to 32 +/- 4 microg/L in normal subjects (n = 10) (P < 0.001). Mean urea reduction was 62% while the reduction ratio for MMA was 43% (P < 0.01). MMA levels rebounded in the 1 hour post-dialytic period to 85% of baseline, whereas urea levels rebounded only to 47% of baseline. MMA had a much larger calculated volume of distribution compared to urea, consistent with intracellular sequestration. Measures of intra-red blood cell (RBC) MMA concentrations confirmed greater levels in RBCs than in plasma with a ratio of 4.9:1. Because of the intracellular sequestration of MMA, we calculated its clearance using that amount removed from whole blood. Clearances for urea averaged 222 +/- 41 ml/min and for MMA 121 +/- 14 ml/min, while plasma clearance for creatinine was 162 +/- 20 ml/min (P < 0.01, for all differences). Using in vitro dialysis, in the absence of RBCs, solute clearance rates were similar: 333 +/- 6, 313 +/- 8 and 326 +/- 4 ml/min for urea, creatinine and MMA, respectively. These findings suggest that the lower MMA clearance relative to creatinine in vivo is a result of MMA movement into RBCs within the dialyser blood path diminishing its removal by dialysis. CONCLUSION: In conclusion, we find that, in conventional haemodialysis, MMA is not cleared as efficiently as urea or creatinine and raise the possibility that RBCs may limit its dialysis not merely by failing to discharge it, but by further sequestering it as blood passes through the dialyser. PMID- 20019017 TI - Inhibition of tyrosine kinases by sunitinib associated with focal segmental glomerulosclerosis lesion in addition to thrombotic microangiopathy. AB - Sunitinib is an orally administered inhibitor of tyrosine kinases and has become the standard of care for many patients with metastatic renal cell carcinoma. Its use has been associated with renal toxicity in some patients. We report a patient with a metastatic clear-cell renal carcinoma who showed arterial hypertension, nephrotic syndrome and azotaemia 10 months after treatment with sunitinib. The renal biopsy revealed focal segmental glomerulosclerosis (FSGS) in addition to thrombotic microangiopathy (TMA), and the complete syndrome disappeared 6 months after sunitinib withdrawal. To our knowledge, this is the first case of FSGS associated to TMA secondary to sunitinib treatment. We discuss the possible glomerular pathomechanism. PMID- 20019018 TI - Primary renal disease in young adults with renal failure. AB - BACKGROUND: No specific data have been published on primary renal disease (PRD) in young adults with end-stage renal failure (ESRF). For children, congenital abnormalities of the kidney and urinary tract (CAKUT) account for 50% of renal failure and other congenital and familial disease comprise 20%. This remains true for teenage children in paediatric registries. METHODS: To investigate the causes of ESRF in young adults, the UK Renal Registry data for the period 2000-2006 have been reviewed and PRD reported for all aged 18-39 years. For comparison, US Renal Data System (USRDS) results are available for age groups 0-19, 20-29 and 30-39 years. These data are also compared with data reported by the British Association of Paediatric Nephrology (BAPN). RESULTS: For the UK, there is a rise in the rate of 'aetiology uncertain' from 6% at 12-15 years to 21% by 18-21 years. This figure of 21% remains constant for the older patients in their third and fourth decades and can be increased by at least 5% by adding 'glomerulonephritis; histologically examined but unspecified'; but these figures compare with unknown rates of 36% for the US age group 20-29 years. In the UK, for those 18-21 years, 'glomerulonephritis' accounts for 28%, when 'Alport's disease' (6.5%) and 'unspecified' (4.5%) are excluded, which compares with age 12-15 of 26%. At age 18-21 years in the UK, there is a sharp decline in all CAKUT (26%) when compared with the BAPN incidence for the 12-15 age group of 45%. For those in their third decade, diabetes accounts for 14-18% of diagnoses, distorting our ability to compare data by percentage. CONCLUSIONS: These young adult data in the UK are consistent with the hypothesis that many of the undiagnosed cases must be CAKUT or tubular disease. PMID- 20019019 TI - Atypical calciphylaxis in a patient receiving warfarin then resolving with cessation of warfarin and application of hyperbaric oxygen therapy. AB - Calciphylaxis is a rare, usually fatal vasculopathic disorder characterized by cutaneous ischemia and necrosis due to calcification of arterioles. Although calciphylaxis is most frequently associated with end-stage renal disease (ESRD) and secondary hyperparathyroidism, it has been reported infrequently among patients on warfarin. No standard treatment has been established for atypical calciphylaxis; however, a potentially beneficial treatment is hyperbaric oxygen therapy (HBOT). A high degree of clinical suspicion, early diagnosis, and understanding the pathophysiology of this disease promotes the optimal management of this extremely morbid and often fatal condition. CASE REPORT: We present a 63 year-old Polynesian woman with biopsy-proven calciphylaxis in the absence of ESRD or elevated serum calcium levels while taking warfarin. Therapeutic dose enoxaparin was substituted for warfarin and she received 40 sessions of HBOT during which lower extremity ulcers resolved. DISCUSSION: Warfarin has been implicated when calciphylaxis presents in an atypical fashion. No guidelines exist for treatment of atypical calciphylaxis in the setting of concomitant warfarin therapy. Up to 80% of calciphylaxis patients die within 1 year of diagnosis. Our patient was changed to low-molecular-weight heparin and received HBOT. CONCLUSION: We present what we believe is the first case of atypical calciphylaxis thought to be attributable to warfarin treated with a therapeutic substitution of anticoagulant and HBOT leading to resolution of cutaneous lesions. PMID- 20019020 TI - The ''critical thrombosis period'' in major orthopedic surgery: when to start and when to stop prophylaxis. AB - Patients undergoing major orthopedic surgery are at high venous thromboembolism (VTE) risk, with morbid and potentially fatal consequences. Anticoagulant VTE prophylaxis reduces rates of postoperative deep vein thrombosis by up to 60% to 70% in these patients. Therefore, pharmacological prophylaxis with low-molecular weight heparins (LMWHs), vitamin K antagonists, or fondaparinux is recommended by current guidelines. However, there remains an ongoing debate regarding when to initiate and the optimal duration for prophylaxis. Here, we discuss the mechanisms underlying thrombus formation in patients undergoing major orthopedic surgery, and we review the current literature on the benefit-to-risk ratio associated with preoperative and postoperative initiation of thromboprophylaxis and also the benefit-to-risk ratio in cases of neuraxial anesthesia. We also discuss the duration of postoperative VTE risk following major orthopedic surgery and assess the ''critical thrombosis period'' when prophylaxis should be provided. Current literature reflects the need to balance the improved efficacy of initiating prophylaxis close to the surgery with increased risk of perioperative bleeding. Evidence from pathology, epidemiology, and clinical studies suggests the risk period for VTE begins at surgery and extends well beyond hospitalization-a crucial issue when considering how long to give prophylaxis-and, in the case of total hip arthroplasty, for at least 3 months after surgery. Literature supports the greater use of ''just-in-time'' thromboprophylaxis initiation and after-discharge continuation of optimal prophylaxis in orthopedic surgery patients. Providing optimal thromboprophylaxis throughout the critical thrombosis period where a patient is at VTE risk will ensure the best reductions in VTE-related morbidity and mortality. PMID- 20019021 TI - T-cell lymphoma and pigtail catheter drainage of a massive paraneoplastic pleuro pericardial effusion in a child. PMID- 20019022 TI - Contrast echocardiography guidance for alcohol septal ablation of hypertrophic obstructive cardiomyopathy. PMID- 20019023 TI - A sword-like foreign body lodged in the ventricular septum: a rare complication of percutaneous vertebroplasty. PMID- 20019024 TI - The risk of tendon xanthomas in familial hypercholesterolaemia is influenced by variation in genes of the reverse cholesterol transport pathway and the low density lipoprotein oxidation pathway. AB - AIMS: The presence of tendon xanthomas is a marker of high risk of cardiovascular disease (CVD) among patients with familial hypercholesterolaemia (FH). Therefore, xanthomas and atherosclerosis may result from the same pathophysiological mechanisms. Reverse cholesterol transport (RCT) and low-density lipoprotein (LDL) oxidation are pathophysiological pathways of atherosclerosis, and it is well established that genetic variation in these pathways influences CVD risk. We therefore determined whether genetic variation in these pathways is also associated with the occurrence of tendon xanthomas in FH patients. METHODS AND RESULTS: Four genetic variants in each pathway were genotyped in 1208 FH patients. We constructed a gene-load score for both pathways. The odds of xanthomas increased with the number of the risk alleles in the RCT pathway (OR 1.21, 95% CI 1.08-1.36, P(trend) = 0.0014). Similarly, higher numbers of risk alleles in the LDL oxidation pathway were associated with the presence of xanthomas (OR 1.24, 95% CI 1.08-1.41, P(trend) = 0.0015). CONCLUSION: The presence of tendon xanthomas in FH patients is associated with genetic variation in the RCT and LDL oxidation pathways. These results support the hypothesis that xanthomas and atherosclerosis share pathophysiological mechanisms. PMID- 20019025 TI - The yield of risk stratification for sudden cardiac death in hypertrophic cardiomyopathy myosin-binding protein C gene mutation carriers: focus on predictive screening. AB - AIMS: We investigated the presence of a clinical diagnosis of hypertrophic cardiomyopathy (HCM) and of risk factors for sudden cardiac death (SCD) at the first cardiological evaluation after predictive genetic testing in asymptomatic carriers of an MYBPC3 gene mutation. METHODS AND RESULTS: Two hundred and thirty five mutation carriers were cardiologically evaluated on the presence of HCM and risk factors. A clinical diagnosis of HCM was made in 53 carriers (22.6%). Disease penetrance at 65 years was incomplete for all types of MYBPC3 gene mutations. Women were affected less often than men (15 and 32% respectively, P = 0.003) and disease penetrance was lower in females than in males (13 and 30% at 50 years, respectively, P = 0.024). One risk factor was present in 87 carriers and 9 had two or more risk factors. Twenty-five carriers (11%) with one or more risk factors and manifest HCM could be at risk for SCD. CONCLUSION: At first cardiological evaluation almost one-quarter of asymptomatic carriers was diagnosed with HCM. Risk factors for SCD were frequently present and 11% of carriers could be at risk for SCD. Predictive genetic testing in HCM families and frequent cardiological evaluation on the presence of HCM and risk factors for SCD are justified until advanced age. PMID- 20019026 TI - Does physical abuse in early childhood predict substance use in adolescence and early adulthood? AB - Prospective longitudinal data from 585 families were used to examine parents' reports of child physical abuse in the first 5 years of life as a predictor of substance use at ages 12, 16, and 24. Path analyses revealed that physical abuse in the first 5 years of life predicted subsequent substance use for females but not males. We found a direct effect of early physical abuse on girls'substance use at age 12 and indirect effects on substance use at age 16 and age 24 through substance use at age 12. For boys, age 12 substance use predicted age 16 substance use, and age 16 substance use predicted age 24 substance use, but physical abuse in the first 5 years of life was unrelated to subsequent substance use. These findings suggest that for females, a mechanism of influence of early physical abuse on substance use into early adulthood appears to be through precocious initiation of substance use in early adolescence. PMID- 20019027 TI - Left ventricular volumetry in healthy children and adolescents: comparison of two different real-time three-dimensional matrix transducers with cardiovascular magnetic resonance. AB - AIMS: To assess the accuracy of different hardware and software settings for left ventricular (LV) volume quantification in children using real-time three dimensional echocardiography (RT3DE). METHODS AND RESULTS: The impact of different matrix transducers (IE 33, X3-1 and VIVID 7, V3) and quantification software settings [TOMTEC; contour-finding activity (tCFA) values ranging from 30 to 70 U] on the accuracy of LV indices was tested in 24 healthy children/adolescents (median = 12.6 years) and 25 paediatric patients with Tetralogy-of-Fallot (TOF) (median = 7.3 years) with abnormally shaped ventricles. RT3DE was compared with cardiovascular magnetic resonance (CMR) volumetry as reference. Best agreement (Bland-Altman analysis) was achieved using a tCFA value of 30 U. Applying the V3 device, end-diastolic volume (EDV) and end-systolic volume (ESV) were underestimated by 14.8 +/- 10.6% (mean +/- SD) and 11.2 +/- 16.3%, respectively (r = 9.42, P < 0.001 and r = 0.937, P = 0.003); with the X3-1 system 24.2 +/- 11.0 and 14.6 +/- 15.2%, respectively (r = 0.951, P < 0.001 and r = 0.912, P = 0.001). Negligible differences <1% (P = n.s.) between both transducers were detected applying a tCFA value of 70 U but with significant underestimation (EDV: approximately 35%, P < 0.001; ESV: approximately 26%, P < 0.001) compared with CMR. EDV and ESV of TOF patients were underestimated by 3.2 +/- 15.4 and 8.1 +/- 22.6%, respectively. Intra- and interobserver variability was <4%. CONCLUSION: In contrast to recommendations of the manufacturer, data sets from both RT3DE transducers showed acceptable agreement to CMR for volumetric parameters only for low tCFA. Fine-tuning of software settings is mandatory to improve accuracy. PMID- 20019029 TI - Continuous coronary perfusion in redo aortic valve replacement following prior coronary surgery; an old trick for new dogs? AB - The optimal myocardial protection method for aortic valve replacement in the setting of prior coronary artery bypass surgery remains a subject of debate. Protection is particularly challenging when a patent pedicled internal thoracic artery graft supplies a proximally obstructed left anterior descending artery. Herein, we describe a modification of an old technique; continuous coronary perfusion, which can be used in selected, anatomically suitable cases. PMID- 20019028 TI - A modular IgG-scFv bispecific antibody topology. AB - Here we present a bispecific antibody (bsAb) format in which a disulfide stabilized scFv is fused to the C-terminus of the light chain of an IgG to create an IgG-scFv bifunctional antibody. When expressed in mammalian cells and purified by one-step protein A chromatography, the bsAb retains parental affinities of each binding domain, exhibits IgG-like stability and demonstrates in vivo IgG like tumor targeting and blood clearance. The extension of the C-terminus of the light chain of an IgG with an scFv or even a smaller peptide does appear to disrupt disulfide bond formation between the light and heavy chains; however, this does not appear to affect binding, stability or in vivo properties of the IgG. Thus, we demonstrate here that the light chain of an IgG can be extended with an scFv without affecting IgG function and stability. This format serves as a standardized platform for the construction of functional bsAbs. PMID- 20019030 TI - Removal of an external stent of the bronchus. AB - We report a case of a patient who underwent removal of an external stent of the bronchus. She had undergone external stenting for tracheobronchomalacia. After removal of the external stent, the bronchus was just as widely patent as preoperative state. PMID- 20019031 TI - Necrotizing fasciitis of the chest wall. AB - Necrotizing fasciitis (NF) is an uncommon infection caused by microorganisms called 'flesh eating bacteria'. It remains a life-threatening condition associated with high mortality rate. Its location to the chest wall is exceptional. Herein, we report the case of a 39-year-old female, without comorbidity, presenting a NF of the chest wall complicating an empyema. We describe the surgical management with a three-step procedure: antibiotherapy debridement, vacuum-assisted closure and delayed surgical reconstruction. PMID- 20019032 TI - Grip strength and cardiovascular drug use in older people: findings from the Hertfordshire Cohort Study. AB - BACKGROUND: reduced grip strength is associated with adverse health consequences, and there is interest in identifying modifiable influences. Cardiovascular drugs are commonly used by older people, but their effect on muscle strength is unclear. METHODS: we investigated associations between cardiovascular drug use and grip strength among 1,572 men and 1,415 women, aged 59-73, who participated in the Hertfordshire Cohort Study. RESULTS: Forty-five percent of participants were taking a cardiovascular drug. Furosemide was associated with average decreases in grip strength of 3.15 kg (95% confidence interval [CI] 0.90, 5.39, P < 0.01) among men and 2.35 kg (95% CI 0.93, 3.77, P < 0.01) among women after adjustment for age and height. Corresponding differences for nitrates were 1.84 kg (95% CI 0.29, 3.39, P = 0.02) among men and 3.66 kg (95% CI 1.99, 5.33, P < 0.01) among women. Calcium channel blockers and fibrates were associated with reduced grip among women. Statins were not associated with grip. The associations between grip strength and nitrate use in men and nitrate and fibrate use in women were robust to additional adjustment for co-morbidity. CONCLUSIONS: use of some cardiovascular drugs is associated with reduced grip strength in older people. These findings have potential implications for the functional ability of older people treated with these drugs. PMID- 20019033 TI - eComment: Pericardiocentesis followed by intrapericardial cisplatin administration in patients with neoplastic pericarditis. PMID- 20019034 TI - eComment: Clamping a calcified aorta: note of caution. PMID- 20019035 TI - eComment: Cross-clamping the heavily calcified ascending aorta after a preoperative computed tomography evaluation. PMID- 20019036 TI - eComment: Re: Right or left anterolateral minithoracotomy for repair of congenital ventricular septal defects in adult patients. PMID- 20019037 TI - eComment: A limited antero-lateral minithoracotomy for congenital ventricular septal defects repair in adult patients. PMID- 20019038 TI - eComment: Leg wound morbidities after saphenous vein harvesting techniques. Which is better? PMID- 20019039 TI - eComment: Influence of 19 mm size aortic valve substitutes on long-term survival. PMID- 20019040 TI - eComment: Controversies on the antiplatelet therapy before coronary artery bypass grafting surgery. PMID- 20019041 TI - eComment: Cardiopulmonary bypass without exchange transfusion in sickle cell disease--an update. PMID- 20019042 TI - eComment: Incomplete left ventricular reverse remodeling after revascularization of anomalous left coronary artery from the pulmonary artery. PMID- 20019043 TI - eComment: Re: Cardiac stunning in the clinic: the full picture. PMID- 20019044 TI - eComment: Sternal microcirculation following pedicled or skeletonized internal thoracic artery harvesting. PMID- 20019045 TI - eComment: Congenital direct communication between the right pulmonary artery and the left atrium: anatomic variations and surgical experience. PMID- 20019046 TI - eComment: Are low molecular weight heparin effective in mechanical valve prosthesis anticoagulation during pregnancy? PMID- 20019047 TI - eComment: Cardiac operation during pregnancy: what is the appropriate management? PMID- 20019048 TI - eComment: Re: Mechanical prosthetic mitral valve thrombosis in a first trimester pregnant woman. PMID- 20019049 TI - eComment: Supradiaphragmatic ligation of the thoracic duct for prevention of postoperative chylothorax. PMID- 20019050 TI - Identification of Mac-2-binding protein as a putative marker of neuroendocrine tumors from the analysis of cell line secretomes. AB - Neuroendocrine tumors (NETs) can arise from a variety of organs. They can vary widely in clinical behavior; consequently, optimizing their treatment plan can be problematic. NETs display diverse tumor biology; however, most secrete peptides such as chromogranin A into the circulation, consistent with their neuroendocrine origin. In this study, we sought to identify other potential markers for NETs by analyzing the secreted proteomes of three neuroendocrine cell lines. BON-1, NCI H727, and SHP-77 cells were grown in serum-free media, and the secreted proteins were separated by SDS-PAGE and identified by LC-MS/MS. We identified 205 proteins of which 61 were secreted by two or more of the cell lines and 19 were secreted by all three lines. Mac-2-binding protein (Mac-2BP) was found to be secreted by all three cell lines, and this was confirmed by Western blotting. Immunohistochemical analysis found 29 of 33 NET cases from different primary sites to be positive for Mac-2BP. Serum Mac-2BP was significantly elevated in NET patients compared with healthy controls (p < 0.001). This study demonstrated that analysis of the secreted proteomes of neuroendocrine cell lines can identify potential biomarkers for NET. Initial assessment showed that serum Mac-2BP is significantly elevated in patients with NET and is expressed by the majority of NET tissues. PMID- 20019051 TI - Neuroproteomics approaches to decipher neuronal regeneration and degeneration. AB - Given the complexity of brain and nerve tissues, systematic approaches are essential to understand normal physiological conditions and functional alterations in neurological diseases. Mass spectrometry-based proteomics is increasingly used in neurosciences to determine both basic and clinical differential protein expression, protein-protein interactions, and post translational modifications. Proteomics approaches are especially useful to understand the mechanisms of nerve regeneration and degeneration because changes in axons following injury or in disease states often occur without the contribution of transcriptional events in the cell body. Indeed, the current understanding of axonal function in health and disease emphasizes the role of proteolysis, local axonal protein synthesis, and a broad range of post translational modifications. Deciphering how axons regenerate and degenerate has thus become a postgenomics problem, which depends in part on proteomics approaches. This review focuses on recent proteomics approaches designed to uncover the mechanisms and molecules involved in neuronal regeneration and degeneration. It emerges that the principal degenerative mechanisms converge to oxidative stress, dysfunctions of axonal transport, mitochondria, chaperones, and the ubiquitin-proteasome systems. The mechanisms regulating nerve regeneration also impinge on axonal transport, cytoskeleton, and chaperones in addition to changes in signaling pathways. We also discuss the major challenges to proteomics work in the nervous system given the complex organization of the brain and nerve tissue at the anatomical, cellular, and subcellular levels. PMID- 20019052 TI - A robust error model for iTRAQ quantification reveals divergent signaling between oncogenic FLT3 mutants in acute myeloid leukemia. AB - The FLT3 receptor tyrosine kinase plays an important role in normal hematopoietic development and leukemogenesis. Point mutations within the activation loop and in frame tandem duplications of the juxtamembrane domain represent the most frequent molecular abnormalities observed in acute myeloid leukemia. Interestingly these gain-of-function mutations correlate with different clinical outcomes, suggesting that signals from constitutive FLT3 mutants activate different downstream targets. In principle, mass spectrometry offers a powerful means to quantify protein phosphorylation and identify signaling events associated with constitutively active kinases or other oncogenic events. However, regulation of individual phosphorylation sites presents a challenging case for proteomics studies whereby quantification is based on individual peptides rather than an average across different peptides derived from the same protein. Here we describe a robust experimental framework and associated error model for iTRAQ-based quantification on an Orbitrap mass spectrometer that relates variance of peptide ratios to mass spectral peak height and provides for assignment of p value, q value, and confidence interval to every peptide identification, all based on routine measurements, obviating the need for detailed characterization of individual ion peaks. Moreover, we demonstrate that our model is stable over time and can be applied in a manner directly analogous to ubiquitously used external mass calibration routines. Application of our error model to quantitative proteomics data for FLT3 signaling provides evidence that phosphorylation of tyrosine phosphatase SHP1 abrogates the transformative potential, but not overall kinase activity, of FLT3-D835Y in acute myeloid leukemia. PMID- 20019054 TI - Emphysematous pyelonephritis, ureteritis and cystitis in a diabetic patient. PMID- 20019053 TI - DtaRefinery, a software tool for elimination of systematic errors from parent ion mass measurements in tandem mass spectra data sets. AB - Hybrid two-stage mass spectrometers capable of both highly accurate mass measurement and high throughput MS/MS fragmentation have become widely available in recent years, allowing for significantly better discrimination between true and false MS/MS peptide identifications by the application of a relatively narrow window for maximum allowable deviations of measured parent ion masses. To fully gain the advantage of highly accurate parent ion mass measurements, it is important to limit systematic mass measurement errors. Based on our previous studies of systematic biases in mass measurement errors, here, we have designed an algorithm and software tool that eliminates the systematic errors from the peptide ion masses in MS/MS data. We demonstrate that the elimination of the systematic mass measurement errors allows for the use of tighter criteria on the deviation of measured mass from theoretical monoisotopic peptide mass, resulting in a reduction of both false discovery and false negative rates of peptide identification. A software implementation of this algorithm called DtaRefinery reads a set of fragmentation spectra, searches for MS/MS peptide identifications using a FASTA file containing expected protein sequences, fits a regression model that can estimate systematic errors, and then corrects the parent ion mass entries by removing the estimated systematic error components. The output is a new file with fragmentation spectra with updated parent ion masses. The software is freely available. PMID- 20019056 TI - Waldenstrom's syndromes. PMID- 20019057 TI - Management of open fractures of the lower limb. PMID- 20019058 TI - Another five year plan for the NHS. PMID- 20019059 TI - FoldAmyloid: a method of prediction of amyloidogenic regions from protein sequence. AB - MOTIVATION: Amyloidogenic regions in polypeptide chains are very important because such regions are responsible for amyloid formation and aggregation. It is useful to be able to predict positions of amyloidogenic regions in protein chains. RESULTS: Two characteristics (expected probability of hydrogen bonds formation and expected packing density of residues) have been introduced by us to detect amyloidogenic regions in a protein sequence. We demonstrate that regions with high expected probability of the formation of backbone-backbone hydrogen bonds as well as regions with high expected packing density are mostly responsible for the formation of amyloid fibrils. Our method (FoldAmyloid) has been tested on a dataset of 407 peptides (144 amyloidogenic and 263 non amyloidogenic peptides) and has shown good performance in predicting a peptide status: amyloidogenic or non-amyloidogenic. The prediction based on the expected packing density classified correctly 75% of amyloidogenic peptides and 74% of non amyloidogenic ones. Two variants (averaging by donors and by acceptors) of prediction based on the probability of formation of backbone-backbone hydrogen bonds gave a comparable efficiency. With a hybrid-scale constructed by merging the above three scales, our method is correct for 80% of amyloidogenic peptides and for 72% of non-amyloidogenic ones. Prediction of amyloidogenic regions in proteins where positions of amyloidogenic regions are known from experimental data has also been done. In the proteins, our method correctly finds 10 out of 11 amyloidogenic regions. AVAILABILITY: The FoldAmyloid server is available at http://antares.protres.ru/fold-amyloid/. PMID- 20019060 TI - Better access to health care facilities is needed to further reduce malaria. PMID- 20019062 TI - Childhood should be alcohol free, says England's chief medical officer. PMID- 20019063 TI - Phosphorylation of Exo1 modulates homologous recombination repair of DNA double strand breaks. AB - DNA double-strand break (DSB) repair via the homologous recombination pathway is a multi-stage process, which results in repair of the DSB without loss of genetic information or fidelity. One essential step in this process is the generation of extended single-stranded DNA (ssDNA) regions at the break site. This ssDNA serves to induce cell cycle checkpoints and is required for Rad51 mediated strand invasion of the sister chromatid. Here, we show that human Exonuclease 1 (Exo1) is required for the normal repair of DSBs by HR. Cells depleted of Exo1 show chromosomal instability and hypersensitivity to ionising radiation (IR) exposure. We find that Exo1 accumulates rapidly at DSBs and is required for the recruitment of RPA and Rad51 to sites of DSBs, suggesting a role for Exo1 in ssDNA generation. Interestingly, the phosphorylation of Exo1 by ATM appears to regulate the activity of Exo1 following resection, allowing optimal Rad51 loading and the completion of HR repair. These data establish a role for Exo1 in resection of DSBs in human cells, highlighting the critical requirement of Exo1 for DSB repair via HR and thus the maintenance of genomic stability. PMID- 20019064 TI - DNA dynamics play a role as a basal transcription factor in the positioning and regulation of gene transcription initiation. AB - We assess the role of DNA breathing dynamics as a determinant of promoter strength and transcription start site (TSS) location. We compare DNA Langevin dynamic profiles of representative gene promoters, calculated with the extended non-linear PBD model of DNA with experimental data on transcription factor binding and transcriptional activity. Our results demonstrate that DNA dynamic activity at the TSS can be suppressed by mutations that do not affect basal transcription factor binding-DNA contacts. We use this effect to establish the separate contributions of transcription factor binding and DNA dynamics to transcriptional activity. Our results argue against a purely 'transcription factor-centric' view of transcription initiation, suggesting that both DNA dynamics and transcription factor binding are necessary conditions for transcription initiation. PMID- 20019065 TI - Cell cycle-dependent variations in protein concentration. AB - Computational modeling of biological systems has become an effective tool for analyzing cellular behavior and for elucidating key properties of the intricate networks that underlie experimental observations. While most modeling techniques rely heavily on the concentrations of intracellular molecules, little attention has been paid to tracking and simulating the significant volume fluctuations that occur over each cell division cycle. Here, we use fluorescence microscopy to acquire single cell volume trajectories for a large population of Saccharomyces cerevisiae cells. Using this data, we generate a comprehensive set of statistics that govern the growth and division of these cells over many generations, and we discover several interesting trends in their size, growth and protein production characteristics. We use these statistics to develop an accurate model of cell cycle volume dynamics, starting at cell birth. Finally, we demonstrate the importance of tracking volume fluctuations by combining cell division dynamics with a minimal gene expression model for a constitutively expressed fluorescent protein. The significant oscillations in the cellular concentration of a stable, highly expressed protein mimic the observed experimental trajectories and demonstrate the fundamental impact that the cell cycle has on cellular functions. PMID- 20019066 TI - Mutations in the perforin gene can be linked to macrophage activation syndrome in patients with systemic onset juvenile idiopathic arthritis. AB - OBJECTIVE: Macrophage activation syndrome (MAS) in systemic onset juvenile idiopathic arthritis (SoJIA) is considered to be an acquired form of familial haemophagocytic lymphohistiocytosis (fHLH). FHLH is an autosomal recessive disorder, characterized by diminished NK cell function and caused by mutations in the perforin gene (PRF1) in 20-50% of patients. Interestingly, SoJIA patients display decreased levels of perforin in NK cells and diminished NK cell function as well. Here, we analysed PRF1 and its putative promoter in SoJIA patients with or without a history of MAS. METHODS: DNA of 56 SoJIA patients (41 Italian and 15 Dutch) was isolated. Of these, 15 (27%) had a confirmed history of MAS. We sequenced PRF1 and 1.5 kb of the 5'-upstream region. DNA sequence variations in the promoter region were functionally tested in transfection experiments using a human NK cell line. RESULTS: We detected a previously undescribed sequence variation (-499 C > T) in the promoter of PRF1 in 18% of the SoJIA patients. However, transfection experiments did not show functional implications of this variation. Secondly, we found that 11 of 56 (20%) SoJIA patients were heterozygous for missense mutations in PRF1. In particular, we found a high prevalence of the Ala91Val mutation, a variant known to result in defective function of perforin. Interestingly, the prevalence of Ala91Val in SoJIA patients with a history of MAS (20%) was increased compared with SoJIA patients without MAS (9.8%). One SoJIA patient, heterozygous for Ala91Val, showed profound decreased perforin levels at the time of MAS. CONCLUSIONS: These findings suggest that PRF1 mutations play a role in the development of MAS in SoJIA patients. PMID- 20019067 TI - Ultrasound-guided sacroiliac joint injection in patients with established sacroiliitis: precise IA injection verified by MRI scanning does not predict clinical outcome. AB - OBJECTIVE: IA injections of SIJs with corticosteroids are often performed in patients suffering from low back pain due to active sacroiliitis. However, SIJ injections are technically demanding, and therefore the clinical outcome of ultrasound-guided corticosteroid SIJ injections was analysed in relation to the accuracy of the injection. METHODS: Ultrasound-guided injections were performed with 40 mg triamcinolone and 0.78 mg gadolinium in 20 SIJ of 14 consecutive patients suffering from active sacroiliitis. Immediately following SIJ injection, MRI scanning was initiated to verify the correct placement of the drug. Clinical outcome of the intervention was determined using a numerical pain rating scale (NRS) at Days 1 and 28. RESULTS: Despite ultrasound guidance, only 8 injections (40%) were exactly positioned into the SIJ space, whereas the other 12 injections (60%) missed the SIJ. However, there were no significant differences observed in the clinical outcome between the IA-injected group and the peri-articular injected group. There was similar pain relief observed in both groups 24 h and 28 days following the intervention [IA injection group: mean NRS-baseline: 6.8 (range 4-9), NRS-24 h: 4.3 (range 1-7) and NRS-day 28: 3.5 (range 1-5); peri articular injection group: mean NRS-baseline: 7.0 (range 5-10), NRS-24 h: 4.1 (range 1-10) and NRS-day 28: 4.5 (range 1-8)]. CONCLUSION: These results demonstrate that IA SIJ injections remain technically challenging despite ultrasound guidance. However, peri-articular deposition of triamcinolone appears sufficient for pain and symptom control in patients suffering from active sacroiliitis. PMID- 20019068 TI - Hydrocephalus and subdural effusion with extracranial extension complicating meningitis. AB - Post-meningitic subdural effusion does not extend extracranially. To our knowledge, the association of hydrocephalus and extracranial extension of a subdural effusion has not been previously reported. We present a case of an 8 year-old boy who presented with progressive head enlargement and worsening pulsatile peri-orbital swelling of 1 year duration. Cranial computerized tomographic scan confirmed communicating hydrocephalus and bilateral subdural effusion worse on the right side. The patient had a ventriculo-peritoneal shunt, with resolution of the subdural effusion and his hydrocephalus. PMID- 20019069 TI - Performance of Pediatric Risk of Mortality, Pediatric Index of Mortality and PIM2 in term Chinese neonates. AB - Pediatric Risk of Mortality (PRISM), Pediatric Index of Mortality (PIM) and PIM2 could be applicable to the subset of term neonates has not been well investigated. The purpose of this study is to access and compare the performance of these scoring systems in predicting mortality probability in term Chinese neonates with critical illness. PRISM, PIM and PIM2 scores were calculated prospectively during a 1-year period on 243 neonates admitted to the neonatal intensive care unit (NICU) in the Children's Hospital of Zhejiang University in China. Of these, 36 neonates (14.81%) died in the NICU, while the mortality rates estimated by PRISM, PIM and PIM2 were 16.19, 14.58 and 11.12%, respectively. The area under the receiver-operating characteristic (ROC) curve [95% confidence intervals (CIs)] were 0.834 (0.767-0.902), 0.851 (0.786-0.916) and 0.854 (0.790 0.918) for PRISM, PIM and PIM2, respectively. The Hosmer-Lemeshow test gave a chi square of 1.35 (p = 0.930) for PRISM, 1.03 (p = 0.960) for PIM and 4.58 (p = 0.469) for PIM2. The standardized mortality rates (SMRs) (95% CI) using PRISM, PIM and PIM2 were 0.92 (0.79-1.08), 1.02 (0.88-1.20) and 1.33 (1.13-1.62), respectively. Although PRISM, PIM and PIM2 have displayed good discrimination and calibration in the present setting, PIM is considered as the most accurate and appropriate tool for predicting mortality in the studied NICU. PMID- 20019070 TI - An open trial of gabapentin in acute alcohol withdrawal using an oral loading protocol. AB - AIMS: Anticonvulsants are increasingly being advocated for the treatment of acute alcohol withdrawal syndrome (AWS) to avoid the addictive properties of established medications. Because earlier works showed that moderate gabapentin doses were too low to clearly ameliorate severe AWS, we tested a higher gabapentin entry dose. METHODS: Inpatients (n = 37) with severe alcohol withdrawal symptoms (Clinical Institute Withdrawal Assessment for Alcohol revised (CIWA-AR) score > or =15 points) were given gabapentin 800 mg, and if their symptom score reduced within 2 h, they were termed 'early responders' and were then treated for 2 days with 600 mg gabapentin q.i.d. (i.e. a total of 3200 mg in the first 24 h) before beginning a taper. RESULTS: Twenty-seven (73%) were early responders (baseline CIWA-AR improved from 17.3 +/- 2.6 to 8.0 +/- 3.6 points). In the remaining 10 patients, baseline CIWA-AR deteriorated within 2 h (from 20.1 +/- 4.6 to 21.5 +/- 4.65 points). These patients were switched to clomethiazole (n = 4) or clonazepam (n = 6), which is the usual treatment. Three of the 'early responders' worsened in the next 36 h and were then reclassified and treated as 'non-responders'. Among them, two developed an epileptic seizure. CONCLUSION: Oral 800 mg gabapentin (loaded up to 3200 mg in the first 24 h) is helpful only in reducing less severe and less complicated acute AWS. PMID- 20019071 TI - ADHD and the DRD4 exon III 7-repeat polymorphism: an international meta-analysis. AB - We sought to elucidate the relationship of ADHD (Attention-Deficit Hyperactivity Disorder) to the DRD4 exon III VNTR 7R allele worldwide using analytic techniques and to relate these findings to the field of cultural neuroscience. To focus on a potential moderating role of race/ethnicity, we excluded over 30 papers that have explored the relationship between the DRD4 7R and ADHD but had unclear or lax racial-ethnic inclusion criteria. The papers in this meta-analysis were only included if a single race made up 95% or more of their sample. We searched for and translated papers not published in English, and found a significant difference in the relationship of ADHD and DRD4 7R in people of European Caucasian (Odds ratio 1.635, Z = 3.936, P < 0.00001) and South American (Odds ratio 2.407, Z = 3.317, P = 0.001) descent vs people of Middle Eastern ancestry (Odds ratio 0.717, Z = -2.466; P = 0.014). We also examined the moderating effect of differing ADHD diagnoses, subject recruitment, control recruitment and male to female ratio. Finally, we consider the implications of these data for cultural neuroscience. PMID- 20019072 TI - Variation in orbitofrontal cortex volume: relation to sex, emotion regulation and affect. AB - Sex differences in brain structure have been examined extensively but are not completely understood, especially in relation to possible functional correlates. Our two aims in this study were to investigate sex differences in brain structure, and to investigate a possible relation between orbitofrontal cortex subregions and affective individual differences. We used tensor-based morphometry to estimate local brain volume from MPRAGE images in 117 healthy right-handed adults (58 female), age 18-40 years. We entered estimates of local brain volume as the dependent variable in a GLM, controlling for age, intelligence and whole brain volume. Men had larger left planum temporale. Women had larger ventromedial prefrontal cortex (vmPFC), right lateral orbitofrontal (rlOFC), cerebellum, and bilateral basal ganglia and nearby white matter. vmPFC but not rlOFC volume covaried with self-reported emotion regulation strategies (reappraisal, suppression), expressivity of positive emotions (but not of negative), strength of emotional impulses, and cognitive but not somatic anxiety. vmPFC volume statistically mediated sex differences in emotion suppression. The results confirm prior reports of sex differences in orbitofrontal cortex structure, and are the first to show that normal variation in vmPFC volume is systematically related to emotion regulation and affective individual differences. PMID- 20019074 TI - Myrica rubra Extracts Protect the Liver from CCl(4)-Induced Damage. AB - The relationship between the expression of mitochondrial voltage-dependent anion channels (VDACs) and the protective effects of Myrica rubra Sieb. Et Zucc fruit extract (MCE) against carbon tetrachloride (CCl(4))-induced liver damage was investigated. Pretreatment with 50 mg kg(-1), 150 mg kg(-1) or 450 mg kg(-1) MCE significantly blocked the CCl(4)-induced increase in both serum aspartate aminotransferase (sAST) and serum alanine aminotransferase (sALT) levels in mice (P < .05 or .01 versus CCl(4) group). Ultrastructural observations of decreased nuclear condensation, ameliorated mitochondrial fragmentation of the cristae and less lipid deposition by an electron microscope confirmed the hepatoprotection. The mitochondrial membrane potential dropped from -191.94 +/- 8.84 mV to -132.06 +/- 12.26 mV (P < .01) after the mice had been treated with CCl(4). MCE attenuated CCl(4)-induced mitochondrial membrane potential dissipation in a dose dependent manner. At a dose of 150 or 450 mg kg(-1) of MCE, the mitochondrial membrane potentials were restored (P < .05). Pretreatment with MCE also prevented the elevation of intra-mitochondrial free calcium as observed in the liver of the CCl(4)-insulted mice (P < .01 versus CCl(4) group). In addition, MCE treatment (50-450 mg kg(-1)) significantly increased both transcription and translation of VDAC inhibited by CCl(4). The above data suggest that MCE mitigates the damage to liver mitochondria induced by CCl(4), possibly through the regulation of mitochondrial VDAC, one of the most important proteins in the mitochondrial outer membrane. PMID- 20019073 TI - Culture, gaze and the neural processing of fear expressions. AB - The direction of others' eye gaze has important influences on how we perceive their emotional expressions. Here, we examined differences in neural activation to direct- versus averted-gaze fear faces as a function of culture of the participant (Japanese versus US Caucasian), culture of the stimulus face (Japanese versus US Caucasian), and the relation between the two. We employed a previously validated paradigm to examine differences in neural activation in response to rapidly presented direct- versus averted-fear expressions, finding clear evidence for a culturally determined role of gaze in the processing of fear. Greater neural responsivity was apparent to averted- versus direct-gaze fear in several regions related to face and emotion processing, including bilateral amygdalae, when posed on same-culture faces, whereas greater response to direct- versus averted-gaze fear was apparent in these same regions when posed on other-culture faces. We also found preliminary evidence for intercultural variation including differential responses across participants to Japanese versus US Caucasian stimuli, and to a lesser degree differences in how Japanese and US Caucasian participants responded to these stimuli. These findings reveal a meaningful role of culture in the processing of eye gaze and emotion, and highlight their interactive influences in neural processing. PMID- 20019075 TI - Red Mold Rice Mitigates Oral Carcinogenesis in 7,12-Dimethyl-1,2 Benz[a]anthracene-Induced Oral Carcinogenesis in Hamster. AB - The prevalence of oral tumor has exponentially increased in recent years; however, the effective therapies or prevention strategies are not sufficient. Red mold rice is a traditional Chinese food, and several reports have demonstrated that red mold rice had an anti-tumor effect. However, the possible anti-tumor mechanisms of the red mold rice are unclear. In this study, we examined the anti tumor effect of red mold rice on 7,12-dimethyl-1,2-benz[a]anthracene (DMBA) induced oral tumor in hamster. The ethanol extract of red mold rice (RMRE) treatment significantly decreases the levels of DMBA-induced reactive oxygen species, nitro oxide and prostaglandin E(2) than those of the lovastatin-treated group (P < .001). Moreover, RMRE decreases the formation of oral tumor induced by DMBA. Monacolin K, monascin, ankaflavin or other red mold rice metabolites had been reported to decrease inflammation and oxidative stress and exerted anti tumor effects. Therefore, we evaluated the anti-inflammation and anti-oxidative stress effects of monacolin K, monascin, ankaflavin and citrinin in lipopolysaccharide-treated RAW264.7 cells. We found that RMRE reduced the LPS induced nitrite levels in RAW264.7 cells better than monacolin K, monascin, ankaflavin or citrinin (P < .05). PMID- 20019076 TI - Expression of, and in vivo stressosome formation by, single members of the RsbR protein family in Bacillus subtilis. AB - The Bacillus subtilis stressosome is a 1.8 MDa complex that is the focal point for activating the bacterium's general response to physical stress. In vitro studies demonstrated that the stressosome's core element can be formed from one or more of a family of paralogous proteins (RsbRA, -RB, -RC and -RD) onto which the system's activator protein (RsbT) and its principal inhibitor (RsbS) are bound. The RsbR components of the stressosome are envisioned to be the initial receptors of stress signalling with the stressosome structure itself serving as a device to integrate multiple stress signals for a coordinated response. In the current work, we examine several of the in vivo characteristics of the RsbR family members, including their expression and ability to form stressosomes to regulate sigma(B). Translational fusions of lacZ to each rsbR paralogue revealed that rsbRA, -RB and -RC are expressed at similar levels, which remain relatively constant during growth, ethanol stress and entry into stationary phase. rsbRD, in contrast, is expressed at a level that is only slightly above background during growth, but is induced to 30 % of the rsbRA expression level following ethanol stress. Velocity sedimentation analyses of B. subtilis extracts from strains expressing single rsbR paralogues demonstrated that each incorporates RsbS into fast-sedimenting complexes. However, consistent with rsbRD's lower expression, the RsbRD-dependent RsbS complexes were present at only 20 % of the level of the complexes seen in a wild-type strain. The lower stressosome level in the RsbRD strain is still able to hold RsbT's activity in check, implying that the RsbR/S component of stressosomes is normally in excess for the control of RsbT. Consistent with such a notion, reporter gene and Western blot assays demonstrate that although RsbT is synthesized at the same rate as RsbRA and RsbS, RsbT's ultimate level in growing B. subtilis is only 10 % that of RsbRA. Apparently, RsbT's inherent structure and/or its passage between the stressosome and its activation target compromises its persistence. PMID- 20019078 TI - Genetic adaptation of Pseudomonas aeruginosa during chronic lung infection of patients with cystic fibrosis: strong and weak mutators with heterogeneous genetic backgrounds emerge in mucA and/or lasR mutants. AB - During the chronic lung infection of patients with cystic fibrosis (CF), Pseudomonas aeruginosa can survive for long periods due to adaptive evolution mediated by genetic variation. Hypermutability is considered to play an important role in this adaptive evolution and it has been demonstrated that mutator populations are amplified in the CF lung by hitchhiking with adaptive mutations. Two of the genes that are frequently mutated in isolates from chronic infection are mucA and lasR. Loss-of-function mutations in these genes determine the phenotypic switch to mucoidy and loss of quorum sensing, which are considered hallmarks of chronic virulence. The aims of our study were to investigate (1) the genetic background of the P. aeruginosa subpopulations with non-mutator, weak or strong mutator phenotype and their dynamics during the chronic lung infection, and (2) the time sequence in which the hypermutable, mucoid and quorum-sensing negative phenotypes emerge during chronic lung infection. For these purposes the sequences of mutS, mutL, uvrD, mutT, mutY and mutM anti-mutator genes as well as of mucA and lasR were analysed in 70 sequential P. aeruginosa isolates obtained from the respiratory secretions of 10 CF patients (one to three isolates per time point). Analysis of the genetic background of the mutator phenotype showed that mutS was the most commonly affected gene followed by mutL in isolates with strong mutator phenotype. The mutT, mutY, mutM genes were affected in isolates with low fold-changes in the mutation frequencies compared to the reference strain PAO1. Isolates with non-mutator, weak or strong mutator phenotype were represented at all time points showing co-existence of these subpopulations, which suggests parallel evolution of the various mutators in the different focal niches of infection in the CF lung. Mutations in mucA and lasR occurred earlier than mutations in the anti-mutator genes, showing that hypermutability is not a prerequisite for the acquisition of mucoidy and loss of quorum sensing, considered hallmarks of chronic virulence. Significantly higher mutation rates and MICs of ceftazidime, meropenem and ciprofloxacin were found for isolates collected late (more than 10 years) during the chronic lung infection compared to isolates collected earlier, which suggests an amplification of the mutator subpopulation by hitchhiking with development of antibiotic resistance. Similar evolutionary pathways concordant with adaptive radiation were observed in different clonal lineages of P. aeruginosa from CF patients. PMID- 20019077 TI - Pan-genome analysis provides much higher strain typing resolution than multi locus sequence typing. AB - The most widely used DNA-based method for bacterial strain typing, multi-locus sequence typing (MLST), lacks sufficient resolution to distinguish among many bacterial strains within a species. Here, we show that strain typing based on the presence or absence of distributed genes is able to resolve all completely sequenced genomes of six bacterial species. This was accomplished by the development of a clustering method, neighbour grouping, which is completely consistent with the lower-resolution MLST method, but provides far greater resolving power. Because the presence/absence of distributed genes can be determined by low-cost microarray analyses, it offers a practical, high resolution alternative to MLST that could provide valuable diagnostic and prognostic information for pathogenic bacterial species. PMID- 20019079 TI - Infection of the Circulifer haematoceps cell line Ciha-1 by Spiroplasma citri: the non-insect-transmissible strain 44 is impaired in invasion. AB - Successful transmission of Spiroplasma citri by its leafhopper vector requires a specific interaction between the spiroplasma surface and the insect cells. With the aim of studying these interactions at the cellular and molecular levels, a cell line, named Ciha-1, was established using embryonic tissues from the eggs of the S. citri natural vector Circulifer haematoceps. This is the first report, to our knowledge, of a cell line for this leafhopper species and of its successful infection by the insect-transmissible strain S. citri GII3. Adherence of the spiroplasmas to the cultured Ciha-1 cells was studied by c.f.u. counts and by electron microscopy. Entry of the spiroplasmas into the insect cells was analysed quantitatively by gentamicin protection assays and qualitatively by double immunofluorescence microscopy. Spiroplasmas were detected within the cell cytoplasm as early as 1 h after inoculation and survived at least 2 days inside the cells. Comparing the insect-transmissible GII3 and non-insect-transmissible 44 strains revealed that adherence to and entry into Ciha-1 cells of S. citri 44 were significantly less efficient than those of S. citri GII3. PMID- 20019080 TI - Mitochondrial processing peptidase activity is controlled by the processing of alpha-MPP during development in Dictyostelium discoideum. AB - We investigated the expression of the alpha subunit of the Dictyostelium mitochondrial processing peptidase (Ddalpha-MPP) during development. Ddalpha-MPP mRNA is expressed at the highest levels in vegetatively growing cells and during early development, and is markedly downregulated after 10 h of development. The Ddalpha-MPP protein is expressed as two forms, designated alpha-MPP(H) and alpha MPP(L), throughout the Dictyostelium life cycle. The larger form, alpha-MPP(H), is cleaved to produce the functional alpha-MPP(L) form. We were not able to isolate mutants in which the alpha-mpp gene had been disrupted. Instead, an antisense transformant, alphaA2, expressing alpha-MPP at a lower level than the wild-type AX-3 was isolated to examine the function of the alpha-MPP protein. Development of the alphaA2 strain was normal until the slug formation stage, but the slug stage was prolonged to approximately 24 h. In this prolonged slug stage, only alpha-MPP(H) was present, and alpha-MPP(L) protein and MPP activity were not detected. After 28 h, alpha-MPP(L) and MPP activity reappeared, and normal fruiting bodies were formed after a delay of approximately 8 h compared with normal development. These results indicate that MPP activity is controlled by the processing of alpha-MPP(H) to alpha-MPP(L) during development in Dictyostelium. PMID- 20019081 TI - Candida albicans sphingolipid C9-methyltransferase is involved in hyphal elongation. AB - C9-methylated glucosylceramide is a fungus-specific sphingolipid. This lipid is a major membrane component in the cell and is thought to play important roles in the growth and virulence of several fungal species. To investigate the importance of the methyl branch of the long-chain base in glucosylceramides in pathogenic fungi, we identified and characterized a sphingolipid C9-methyltransferase gene (MTS1, C9-MethylTransferase for Sphingolipid 1) in the pathogenic yeast Candida albicans. The mts1 disruptant lacked (E,E)-9-methylsphinga-4,8-dienine in its glucosylceramides and contained (E)-sphing-4-enine and (E,E)-sphinga-4,8-dienine. Reintroducing the MTS1 gene into the mts1 disruptant restored the synthesis of (E,E)-9-methylsphinga-4,8-dienine in the glucosylceramides. We also created a disruptant of the HSX11 gene, encoding glucosylceramide synthase, which catalyses the final step of glucosylceramide synthesis, in C. albicans and compared this mutant with the mts1 disruptant. The C. albicans mts1 and hsx11 disruptants both had a decreased hyphal growth rate compared to the wild-type strain. The hsx11 disruptant showed increased susceptibility to SDS and fluconazole, similar to a previously reported sld1 disruptant that contained only (E)-sphing-4-enine in its glucosylceramides, suggesting that these strains have defects in their cell membrane structures. In contrast, the mts1 disruptant grew similarly to wild-type in medium containing SDS or fluconazole. These results suggest that the C9-methyl group of a long-chain base in glucosylceramides plays an important role in the hyphal elongation of C. albicans independent of lipid membrane disruption. PMID- 20019082 TI - Metals, minerals and microbes: geomicrobiology and bioremediation. AB - Microbes play key geoactive roles in the biosphere, particularly in the areas of element biotransformations and biogeochemical cycling, metal and mineral transformations, decomposition, bioweathering, and soil and sediment formation. All kinds of microbes, including prokaryotes and eukaryotes and their symbiotic associations with each other and 'higher organisms', can contribute actively to geological phenomena, and central to many such geomicrobial processes are transformations of metals and minerals. Microbes have a variety of properties that can effect changes in metal speciation, toxicity and mobility, as well as mineral formation or mineral dissolution or deterioration. Such mechanisms are important components of natural biogeochemical cycles for metals as well as associated elements in biomass, soil, rocks and minerals, e.g. sulfur and phosphorus, and metalloids, actinides and metal radionuclides. Apart from being important in natural biosphere processes, metal and mineral transformations can have beneficial or detrimental consequences in a human context. Bioremediation is the application of biological systems to the clean-up of organic and inorganic pollution, with bacteria and fungi being the most important organisms for reclamation, immobilization or detoxification of metallic and radionuclide pollutants. Some biominerals or metallic elements deposited by microbes have catalytic and other properties in nanoparticle, crystalline or colloidal forms, and these are relevant to the development of novel biomaterials for technological and antimicrobial purposes. On the negative side, metal and mineral transformations by microbes may result in spoilage and destruction of natural and synthetic materials, rock and mineral-based building materials (e.g. concrete), acid mine drainage and associated metal pollution, biocorrosion of metals, alloys and related substances, and adverse effects on radionuclide speciation, mobility and containment, all with immense social and economic consequences. The ubiquity and importance of microbes in biosphere processes make geomicrobiology one of the most important concepts within microbiology, and one requiring an interdisciplinary approach to define environmental and applied significance and underpin exploitation in biotechnology. PMID- 20019083 TI - Genome-wide analysis of DNA repeats in Burkholderia cenocepacia J2315 identifies a novel adhesin-like gene unique to epidemic-associated strains of the ET-12 lineage. AB - Members of the Burkholderia cepacia complex (Bcc) are respiratory pathogens in patients with cystic fibrosis (CF). Close repetitive DNA sequences often associate with surface antigens to promote genetic variability in pathogenic bacteria. The genome of Burkholderia cenocepacia J2315, a CF isolate belonging to the epidemic lineage Edinburgh-Toronto (ET-12), was analysed for the presence of close repetitive DNA sequences. Among the 422 DNA close repeats, 45 genes potentially involved in virulence were identified and grouped into 12 classes; of these, 13 genes were included in the antigens class. Two trimeric autotransporter adhesins (TAA) among the 13 putative antigens are absent from the other Burkholderia genomes and are clustered downstream of the cci island that is a marker for transmissible B. cenocepacia strains. This cluster contains four adhesins, one outer-membrane protein, one sensor histidine kinase and two transcriptional regulators. By using PCR, we analysed three genes among 47 Bcc isolates to determine whether the cluster was conserved. These three genes were present in the isolates of the ET-12 lineage but absent in all the other members. Furthermore, the BCAM0224 gene was exclusively detected in this epidemic lineage and may serve as a valuable new addition to the field of Bcc diagnostics. The BCAM0224 gene encodes a putative TAA that demonstrates adhesive properties to the extracellular matrix protein collagen type I. Quantitative real-time PCR analysis indicated that BCAM0224 gene expression occurred preferentially for cells grown under high osmolarity, oxygen-limited conditions and oxidative stress. Inactivation of BCAM0224 in B. cenocepacia attenuates the ability of the mutant to promote cell adherence in vitro and impairs the overall bacterial virulence against Galleria mellonella as a model of infection. Together, our data show that BCAM0224 from B. cenocepacia J2315 represents a new collagen-binding TAA with no bacterial orthologues which has an important role in cellular adhesion and virulence. PMID- 20019084 TI - Cytocidal amino acid starvation of Saccharomyces cerevisiae and Candida albicans acetolactate synthase (ilv2{Delta}) mutants is influenced by the carbon source and rapamycin. AB - The isoleucine and valine biosynthetic enzyme acetolactate synthase (Ilv2p) is an attractive antifungal drug target, since the isoleucine and valine biosynthetic pathway is not present in mammals, Saccharomyces cerevisiae ilv2Delta mutants do not survive in vivo, Cryptococcus neoformans ilv2 mutants are avirulent, and both S. cerevisiae and Cr. neoformans ilv2 mutants die upon isoleucine and valine starvation. To further explore the potential of Ilv2p as an antifungal drug target, we disrupted Candida albicans ILV2, and demonstrated that Ca. albicans ilv2Delta mutants were significantly attenuated in virulence, and were also profoundly starvation-cidal, with a greater than 100-fold reduction in viability after only 4 h of isoleucine and valine starvation. As fungicidal starvation would be advantageous for drug design, we explored the basis of the starvation cidal phenotype in both S. cerevisiae and Ca. albicans ilv2Delta mutants. Since the mutation of ILV1, required for the first step of isoleucine biosynthesis, did not suppress the ilv2Delta starvation-cidal defects in either species, the cidal phenotype was not due to alpha-ketobutyrate accumulation. We found that starvation for isoleucine alone was more deleterious in Ca. albicans than in S. cerevisiae, and starvation for valine was more deleterious than for isoleucine in both species. Interestingly, while the target of rapamycin (TOR) pathway inhibitor rapamycin further reduced S. cerevisiae ilv2Delta starvation viability, it increased Ca. albicans ilv1Delta and ilv2Delta viability. Furthermore, the recovery from starvation was dependent on the carbon source present during recovery for S. cerevisiae ilv2Delta mutants, reminiscent of isoleucine and valine starvation inducing a viable but non-culturable-like state in this species, while Ca. albicans ilv1Delta and ilv2 Delta viability was influenced by the carbon source present during starvation, supporting a role for glucose wasting in the Ca. albicans cidal phenotype. PMID- 20019086 TI - Economic evaluation of docetaxel-gemcitabine versus vinorelbine-cisplatin combination as front-line treatment of patients with advanced/metastatic non small-cell lung cancer in Greece: a cost-minimization analysis. AB - BACKGROUND: An economic evaluation was undertaken, alongside a randomized phase III study, to assess docetaxel-gemcitabine (DG) relative to vinorelbine-cisplatin (VC) combination as front-line treatment of patients with advanced/metastatic non small-cell lung cancer. METHODS: No differences were found in efficacy, thus a cost-minimization analysis was carried out. Treatment cost accounts for the administration of first- and second-line chemotherapy, for concomitant medications, for laboratory and biochemical examinations, and for hospitalizations due to adverse events. Unit prices used reflect 2008 and are common among National Health Service hospitals in Greece. RESULTS: The mean total cost of therapy in the DG group [14045 euros, 95% uncertainty interval (UI) 12628 euros-15390 euros], was significantly higher than in the VC group (8143 euros, 95% UI 7314 euros-9067 euros). CONCLUSIONS: Even though the combination VC has similar effectiveness compared with DG in patients with metastatic lung cancer, it is associated with a lower overall treatment cost and hence, it is preferable from an economic perspective. However, it should be noted that although VC is associated with lower cost, it is also more toxic than DG regimen, a significant parameter that should be considered in clinical decisions. PMID- 20019085 TI - G{alpha}5 subunit-mediated signalling requires a D-motif and the MAPK ERK1 in Dictyostelium. AB - The Dictyostelium Galpha5 subunit has been shown to reduce cell viability, inhibit folate chemotaxis and accelerate tip morphogenesis and gene expression during multicellular development. Alteration of the D-motif (mitogen-activated protein kinase docking site) at the amino terminus of the Galpha 5 subunit or the loss of extracellular signal-regulated kinase (ERK)1 diminished the lethality associated with the overexpression or constitutive activation of the Galpha5 subunit. The amino-terminal D-motif of the Galpha5 subunit was also found to be necessary for the reduced cell size, small aggregate formation and precocious developmental gene expression associated with Galpha5 subunit overexpression. This D-motif also contributed to the aggregation delay in cells expressing a constitutively active Galpha5 subunit, but the D-motif was not necessary for the inhibition of folate chemotaxis. These results suggest that the amino-terminal D motif is required for some but not all phenotypes associated with elevated Galpha5 subunit functions during growth and development and that ERK1 can function in Galpha5 subunit-mediated signal transduction. PMID- 20019087 TI - Development of a new G-CSF product based on biosimilarity assessment. AB - BACKGROUND: Zarzio, a new recombinant human granulocyte colony-stimulating factor (filgrastim), was evaluated in healthy volunteers and neutropenic patients in phase I and III studies. PATIENTS AND METHODS: Healthy volunteers in randomized, two-period crossover studies received single- and multiple-dose s.c. injections of 1 microg/kg (n = 24), 2.5 microg/kg (n = 28), 5 microg/kg (n = 28), or 10 microg/kg (n = 40), as well as single-dose i.v. infusions of 5 microg/kg (n = 26), of Zarzio or the reference product (Neupogen). Filgrastim serum levels were monitored; pharmacodynamic parameters were absolute neutrophil count (all studies) and CD34(+) cells (multiple-dose studies). Supportive efficacy and safety data were obtained from an open phase III study in 170 breast cancer patients undergoing four cycles of doxorubicin and docetaxel (Taxotere) chemotherapy, receiving Zarzio (300 or 480 microg) as primary prophylaxis of severe neutropenia. RESULTS: The results of the studies in healthy volunteers confirm the comparability of the test and reference products with respect to their pharmacodynamics and pharmacokinetics. Confidence intervals were within the predefined equivalence boundaries. In the phase III study in breast cancer patients, the administration of Zarzio was efficacious and safe, triggering no immunogenicity. CONCLUSION: The results of these studies demonstrate the biosimilarity of Zarzio with its reference product Neupogen. PMID- 20019088 TI - Bone metastasis and poor performance status are prognostic factors for survival of carcinoma of unknown primary site in patients treated with systematic chemotherapy. AB - BACKGROUND: Cancer of unknown primary site (CUP) generally has a poor prognosis, and there is no established standard therapy. There have been no reports of a prognostic model for CUP patients treated with a single regimen of systemic chemotherapy. METHODS: Univariate and multivariate prognostic factor analysis for overall survival (OS) were conducted retrospectively in 58 consecutive CUP patients treated with carboplatin plus paclitaxel (Taxol) therapy as a first-line treatment. RESULTS: Univariate prognostic factor analysis revealed baseline performance status (PS) of two or more, low serum albumin level, pleural effusion, bone metastasis, and liver metastasis as adverse prognostic factors. Cox proportional hazards analysis showed that poor PS and bone metastasis had the most powerful adverse impact on survival. We developed a prognostic model using those two variables-a good-risk group (PS 0-1 without bone metastasis) and a poor risk group (PS > or =2 or bone metastasis). The poor-risk group showed significantly poorer OS than the good-risk group (1 year OS 36.8% versus 67.1%, P = 0.0003). CONCLUSIONS: Poor PS and bone metastasis were identified as independent adverse prognostic factors in CUP. A simple prognostic model was developed and seems useful for decision making as to whether chemotherapy is indicated for CUP patients. PMID- 20019089 TI - Second cancers after testicular cancer diagnosed after 1980 in Sweden. AB - BACKGROUND: Testicular cancer treatment has become standardized in the 1980s involving radiotherapy preferentially for seminoma and chemotherapy for nonseminoma. The late effects of these therapeutic practices have not been properly evaluated because of the relatively short time since their application. PATIENTS AND METHODS: We conducted a study among 5533 survivors of testicular cancer on the basis of Swedish Family-Cancer Database for which the cancer data were retrieved from the nationwide Cancer Registry. Standardized incidence ratios (SIRs) of second cancer were calculated by comparing with the rates of the first cancers. The follow-up was started in 1980 and carried on through 2006. RESULTS: A total of 370 second cancers (6.7% of all patients) were recorded, more in seminoma than in nonseminoma patients. Second testicular cancer showed an SIR of 29 after seminoma and an SIR of 13 after nonseminoma. A total of 10 discordant sites were increased after seminoma compared with seven sites after nonseminoma. Gastrointestinal tract cancers occurred mainly after seminoma and bladder cancers (SIR 8.6 when diagnosis before age 30) occurred after nonseminoma. CONCLUSIONS: Of the selective affected second tumors, it will be important to confirm the association of bladder cancer with nonseminoma treatment. PMID- 20019090 TI - Prognostic impact of proliferative index determined by quantitative image analysis and the International Prognostic Index in patients with mantle cell lymphoma. AB - BACKGROUND: The proliferative index (PI) is a powerful prognostic factor in mantle cell lymphoma (MCL); however, its utility is hampered by interobserver variability. The mantle cell international prognostic index (MIPI) has been reported to have prognostic importance. In this study, we determined the prognostic value of the PI as determined by quantitative image analysis in MCL. PATIENTS AND METHODS: Eighty-eight patients with adequate tissue were included in this analysis. Patients were treated with one of two treatment programs: sequential therapy with high-dose therapy consolidation or radioimmunotherapy followed by combination chemotherapy with cyclophosphamide, doxorubicin, vincristine and prednisone. Patients were divided into four groups based on PI (<10%, 10%-29.9%, 30%-49.9%, and >50%), and outcomes were analyzed. RESULTS: Thirty percent was identified as the optimal cut-off for PI. By univariate analysis, intensive treatment and a low PI were associated with a superior progression-free survival (PFS); only PI was associated with overall survival. By multivariate analysis, both intensive treatment and PI correlated with PFS. The MIPI had no prognostic impact. CONCLUSIONS: PI is the most important prognostic factor in MCL. The cut-off of 30% is clinically meaningful and can be used to tailor the intensity of therapy in future clinical trials. PMID- 20019091 TI - Measuring morbidity: self-report or health care records? AB - BACKGROUND: Epidemiological surveys often rely on self-report as a measure of morbidity in a population. However, these data can also be extracted from primary care records. OBJECTIVE: To compare morbidity estimates based on self-report with those obtained from primary care records. METHODS: A cross-sectional survey and accompanying medical record review were carried out in all consenting adults aged > or =50 years in three general practices in North Staffordshire, UK. Self reported morbidity was compared with computerized general practice consultation records for the 2 years prior to the survey. RESULTS: Of the 7878 survey responders, 5889 consented to medical record review. Agreement between self reported and consultation data was excellent for diabetes. Agreement between the two sources of data was lower for hypertension, heart problems, chest problems and eyesight problems. It was poor for deafness and falls. CONCLUSIONS: Self report and consultation data provide comparable estimates of the prevalence of specific diagnoses such as diabetes. For other conditions, self-report and consultation records provide different measures of prevalence, and the choice of measure will depend on the morbidity being studied. PMID- 20019092 TI - The EWS/FLI1 oncogenic protein inhibits expression of the Wnt inhibitor DICKKOPF 1 gene and antagonizes beta-catenin/TCF-mediated transcription. AB - Tumours of the Ewing family, which comprise Ewing's sarcoma and peripheral primitive neuroectodermal tumours, are highly aggressive and mostly affect children and adolescents. They are characterized by chromosomal translocations leading to the generation of fusion proteins between EWS (or very rarely FUS) and members of the E-twenty-six (ETS) family of transcription factors that are capable of transforming cells. EWS/FLI1, the most frequent fusion, is thought to cause transformation through activation or repression of specific target genes. We present evidence demonstrating that the Wnt inhibitor and beta-catenin/T-cell factor (TCF)-responsive gene DICKKOPF-1 (DKK-1) is a transcriptional target of EWS/FLI1, which can inhibit both basal and beta-catenin-induced transactivation of the DKK-1 promoter. Moreover, our data indicate that EWS/FLI1 has a more general effect on beta-catenin/TCF-mediated transcription since it can block transactivation of a consensus beta-catenin/TCF reporter construct. Consistently, Ewing tumour cells expressing different EWS/ETS translocations cannot engage beta catenin/TCF-dependent transcription, whereas silencing of EWS/FLI1 restores beta catenin responsiveness in A673 and RD-ES Ewing tumour cells. Accordingly, gene set enrichment analysis shows that beta-catenin/TCF target genes are significantly enriched among genes downregulated by EWS/FLI1 in the Ewing cell line A673. Mechanistically, the inhibitory effect of EWS/FLI1 can be overcome by a constitutively active TCF4 protein (TCF4-VP16). Moreover, EWS/FLI1 binds lymphoid enhancer factor 1, a TCF family member, and interferes with its binding to beta-catenin, which could explain its negative effect on beta-catenin/TCF mediated transcription. Our results show that EWS/FLI1 inhibits both DKK-1 expression as well as beta-catenin/TCF-dependent transcription, which could contribute to progression of tumours of the Ewing family. PMID- 20019093 TI - Recent progress in deciphering the biosynthesis of aspartate-derived amino acids in plants. AB - Plants are either directly or indirectly the source of most of the essential amino acids in animal diets. Four of these essential amino acids-methionine, threonine, isoleucine, and lysine-are all produced from aspartate via a well studied biosynthesis pathway. Given the nutritional interest in essential amino acids, the aspartate-derived amino acid pathway has been the subject of extensive research. Additionally, several pathway enzymes serve as targets for economically important herbicides, and some of the downstream products are biosynthetic precursors for other essential plant metabolites such as ethylene and S adenosylmethionine. Recent and ongoing research on the aspartate-derived family of amino acids has identified new enzyme activities, regulatory mechanisms, and in vivo metabolic functions. Together, these discoveries will open up new possibilities for plant metabolic engineering. PMID- 20019094 TI - Influence of cereal bran supplement on cadmium effects in growing rats. AB - Strategies to diminish cadmium (Cd) absorption are highly desirable especially where Cd exposure due to environmental contamination is still inevitable. Cd toxicity may be influenced by dietary components, such as fiber and minerals. Multimixtures are low-cost cereal bran supplements used in Brazil and in other countries to counteract malnutrition in low-income populations. This study was aimed at evaluating whether multimixture would reduce Cd effects in young rats. Animals received a diet with or without the multimixture plus 0, 5, or 25 mg Cd/kg (control, Cd-5, and Cd-25 groups) during 30 days. The Cd-5 groups were similar to control groups in all parameters analyzed, except in the higher renal Cd concentration. However, the Cd-25 groups had lower biological growth parameters and renal delta-aminolevulinate dehydratase activity, besides higher renal Cd concentration and plasma alanine aminotransferase activity compared to the controls. The multimixture did not prevent Cd effects in the Cd-25 group, but caused a small reduction in renal Cd concentration in the Cd-5 group. Although this multimixture was ineffective to prevent Cd effects at the higher concentration, it seemed to reduce Cd accumulation at the lower Cd dietary concentration, which is similar to levels of human exposure in some polluted areas. PMID- 20019095 TI - Pattern of pediatric poisoning in the east Karadeniz region between 2002 and 2006: increased suicide poisoning. AB - In the present study, 386 patients with the diagnosis of poisoning admitted to the Pediatric Emergency Unit of Farabi Hospital of Medical Faculty of Karadeniz Technical University between January 2002 and December 2006 were retrospectively evaluated with respect to gender, age, cause of poisoning, type of substance used, route of exposure, reason for the intake, signs and symptoms, time of referral to the hospital, hospitalization period, and prognosis. The age group of most poisoning cases was <5 years of age and constituted 51% (n = 197) of all cases. The main toxic agent was drugs (70.2%), followed by foods (8.8%), rodenticides (7%), insecticides/pesticides (4.9%), and carbon monoxide (4.7%). In childhood poisonings, accidental drug poisoning was frequent in toddlers, whereas suicidal poisoning was frequent in adolescents. The suicidal poisoning rate was 23.8% among all poisoning patients, and 98.9% of these patients were adolescents. The suicidal poisoning rates for males and females were 30% and 70%, respectively. An increase in suicidal and inhalation poisonings was observed when compared with previous studies that have been conducted in the same region. The results of the present study suggest that poisonings still represents an important health problem that could be prevented by safe drug storage at home, as well as parental education on adolescence issues, particularly those regarding females. PMID- 20019096 TI - Melanoma in multiple sclerosis treated with natalizumab: causal association or coincidence? AB - We report a relapsing-remitting multiple sclerosis patient who received monthly intravenous natalizumab. After the fifth dose, the patient had a change in a long standing mole. Ten months later, the mole became ulcerative and was ablated. Histological examination identified a spreading melanoma reaching the lower dermis (Clark level IV). Considering that at the moment the incidence of melanoma is estimable as about 5 per 100,000 multiple sclerosis person-years treated with natalizumab, and that, in the general population, the incidence of melanoma per 100,000 person-years is more than 10, we may speculate that the occurrence of melanoma during natalizumab treatment in multiple sclerosis is purely a coincidence. PMID- 20019097 TI - Transient cardiomyopathy as the presenting feature of acute disseminated encephalomyelitis. AB - Brainstem lesions are rarely associated with neurogenic pulmonary oedema (NPO) in multiple sclerosis and other disorders. The exact mechanism for this is unknown. We describe a case of a 15-year-old boy who presented with transient cardiomyopathy and severe acute pulmonary oedema. Several days after his initial presentation he developed an ataxic syndrome with limb, truncal and gait ataxia and nystagmus on primary gaze. Investigations confirmed acute disseminated encephalomyelitis (ADEM). For the first time, we describe a case of transient cardiomyopathy and NPO as the initial manifestation of ADEM. PMID- 20019098 TI - Multiple sclerosis in Sudan: a prospective study of clinical presentation and outcome. PMID- 20019100 TI - Enhancing fraction in glioma and its relationship to the tumoral vascular microenvironment: A dynamic contrast-enhanced MR imaging study. AB - BACKGROUND AND PURPOSE: EnF is a newly described measure of proportional tumor enhancement derived from DCE-MR imaging. The aim of this study was to assess the relationship between EnF and the more established DCE-MR imaging parameters: K(trans), v(e), and v(p). MATERIALS AND METHODS: Forty-two patients with 43 gliomas (16 grade II, 3 grade III, and 24 grade IV) were studied. Imaging included pre- and postcontrast T1-weighted sequences through the lesion and T1 weighted DCE-MR imaging. Parametric maps of EnF, K(trans), v(e), and v(p) were generated. Voxels were classified as enhancing if the IAUC was positive (EnF(IAUC)(60>0)). A threshold of IAUC > 2.5 mmol.s was used to generate EnF(IAUC)(60>2.5). Both measures of EnF were compared with the DCE-MR imaging parameters (K(trans), v(e), and v(p)). RESULTS: In grade II gliomas, EnF(IAUC60>0) and EnF(IAUC60>2.5) correlated with v(p) (R(2) = 0.6245, P < .0005; and R(2) = 0.4727, P = .003) but not with K(trans) or v(e). In grade IV tumors, both EnF(IAUC60>0) and EnF(IAUC60>2.5) correlated with K(trans) (R(2) = 0.3501, P = .001; and R(2) = 0.4699, P < .0005) and v(p) (R(2) = 0.1564, P = .01; and R(2) = 0.2429, P = .007), but not with v(e). Multiple regression analysis showed K(trans) as the only independent correlate of both EnF(IAUC60>0) and EnF(IAUC60>2.5) for grade IV tumors. CONCLUSIONS: This study suggests that in grade II tumors, EnF reflects v(p) and varies due to changes in vascular density. In grade IV gliomas, EnF is affected by K(trans) with secondary associated changes in v(p). PMID- 20019101 TI - Diffusion-weighted MR imaging of intramedullary spinal cord abscess. AB - Early diagnosis and prompt initiation of adequate treatment are essential for clinical outcome in ISCA. We report a case in which DWI provided a more specific diagnosis than conventional MR imaging and allowed differentiation of a ring enhancing lesion from intramedullary tumor. Diagnosis was proved by PCR from CSF (Streptococcus intermedius). Adequate antibiotic treatment was immediately initiated, and the patient recovered completely. PMID- 20019102 TI - Tactile sensory and pain networks in the human spinal cord and brain stem mapped by means of functional MR imaging. AB - BACKGROUND AND PURPOSE: Touch and brush sensory stimuli elicit activity in discriminative touch pathways involving specific regions in the spinal cord and brain stem. However, no study has mapped normal sensory activity noninvasively in healthy humans. The purpose of this study is to map the neuronal activity of sensory input to understand abnormal sensory transmission. MATERIALS AND METHODS: In the present study, spinal fMRI (by using SEEP) was used to map the activity involved with light touch (2 g and 15 g von Frey filaments) and brush stimuli in the brain stem and spinal cords of 8 healthy volunteers. The results were spatially normalized and analyzed with custom-made software. Areas of SEEP activity were identified by using general linear model analysis. RESULTS: The 2 g von Frey filament showed predominant activity in the medulla around the ipsilateral dorsal gracile and cuneate nuclei. The 15 g filament elicited significant activity in the ipsilateral dorsal and contralateral ventral gray matter areas of the spinal cord, areas around the olivary nuclei, pontine reticular formation, periaqueductal gray, and raphe nuclei in the rostral pons and midbrain. The brush stimuli elicited more activity in the medulla around the ipsilateral cuneate and gracile nuclei. CONCLUSIONS: The 2 g filament and brush stimuli activated areas associated with a touch response. The 15 g filament activated areas associated with a pain response. The results from this study identify specific neuronal regions in the brain stem and spinal cord involved in sensory transmission and help understand altered sensory and pain states. PMID- 20019103 TI - MR imaging in multiple sclerosis: review and recommendations for current practice. AB - MR imaging is widely used for the diagnosis and monitoring of patients with MS. Applications and protocols for MR imaging continue to evolve, prompting a need for continual reassessments of the optimal use of this technique in clinical practice. This article provides updated recommendations on the use of MR imaging in MS, based on a review of the trial evidence and personal experiences shared at a recent expert meeting of radiologists and neurologists. PMID- 20019104 TI - Diffusion tensor imaging in hypertrophic olivary degeneration. AB - SUMMARY: We report DTI and fiber tractography findings in a case of hypertrophic olivary degeneration. A 51-year-old man presented with an abnormal gait and visual difficulties. MR imaging showed enlargement of the right medullary olive and a vascular lesion in the right pontine tegmentum. Fiber tractography showed decreased volume of the right central tegmental tract, supporting a diagnosis of HOD. PMID- 20019105 TI - Neuroradiology without benefit of computers: a memoir. PMID- 20019106 TI - Pedicle involvement on MR imaging is common in osteoporotic compression fractures. AB - BACKGROUND AND PURPOSE: Pedicle involvement on MR imaging has been considered specific for malignancy. However, we also noted the findings in many patients with osteoporosis and hypothesized that it is not specific for malignant lesions. The aim of this study was to evaluate the prevalence of pedicle involvement in painful osteoporotic compression fractures and to determine whether the sign is specific for malignancy. MATERIALS AND METHODS: We retrospectively reviewed MR images and CT scans of 152 patients who underwent PV for painful compression fractures. There were 140 patients (225 vertebrae) with osteoporotic fractures and 12 patients (19 vertebrae) with malignant fractures. Three radiologists evaluated the degree and extent of signal-intensity changes of the pedicle on MR imaging by consensus. The CT findings were also evaluated. The chi(2) test was used for statistical analyses. RESULTS: Of the 225 vertebrae of osteoporotic fractures and 19 vertebrae of malignant fractures, pedicle involvement on MR imaging was seen in 144 (64%) and 16 (84.2%) vertebrae, respectively, and there was no statistically significant difference (P = .065). Positive pedicle involvement in osteoporotic fractures was seen in 84 (77%) of 109 vertebrae with early-phase fractures (< or =3 months) and 60 (51.7%) of 116 vertebrae with chronic-phase fractures (>3 months), and this was statistically significant (P < .001). Among 144 osteoporotic vertebrae that showed positive pedicle involvement on MR imaging, 45 (31%) showed pedicle fractures and 55 (38.2%) showed sclerotic change on CT. CONCLUSIONS: Pedicle involvement was seen frequently in patients with osteoporotic compression fractures and was not specific for malignancy in our study group. PMID- 20019107 TI - A prospective trial of 3T and 1.5T time-of-flight and contrast-enhanced MR angiography in the follow-up of coiled intracranial aneurysms. AB - BACKGROUND AND PURPOSE: Endovascularly coiled intracranial aneurysms are increasingly being followed up with noninvasive MRA imaging to evaluate for aneurysm recurrences. It has not been well-established which MRA techniques are best for this application, however. Our aim was to prospectively compare 4 MRA techniques, TOF and CE-MRA at 1.5T and 3T, to a reference standard of DSA in the evaluation of previously endovascularly coiled intracranial aneurysms. MATERIALS AND METHODS: Fifty-eight subjects with 63 previously coiled intracranial aneurysms underwent all 4 MRA techniques within 8 days of DSA. There were 2 outcome variables: coil occlusion class (class 1, complete; class 2, dog ear; class 3, residual neck; class 4, aneurysm filling) and change in degree of occlusion since the previous comparison. Sensitivity and specificity were computed for each MRA technique relative to the reference standard of DSA. Differences among the MRA techniques were evaluated in pair-wise fashion by using the McNemar test. RESULTS: For the detection of any aneurysm remnant, the sensitivity was 85%-90% for all MRA techniques. Sensitivity dropped to 50%-67% when calculated for the detection of only the class 3 and 4 aneurysm remnants, because several class 3 and 4 remnants were misclassified as class 2 by MRA. CE MRA at 1.5T and 3T misclassified fewer of the class 3 and 4 remnants than did TOF MRA at 1.5T, as reflected by the significantly greater sensitivity for larger aneurysm remnants with CE-MRA relative to TOF-MRA at 1.5T (P = .0455 for both comparisons). CONCLUSIONS: CE-MRA is more likely than TOF-MRA to classify larger aneurysm remnants appropriately. We recommend performing both CE-MRA and TOF-MRA in the follow-up of coiled intracranial aneurysms and at 3T if available. PMID- 20019108 TI - Marchiafava-Bignami disease with widespread lesions and complete recovery. AB - MBD is a rare disorder strongly associated with alcoholism. It is characterized pathologically by demyelination and necrosis of the corpus callosum. MBD presents with severe neurologic deficits and significant sequelae developing in most survivors. We report a patient with total clinical recovery. Serial MR imaging demonstrated typical lesions with restricted diffusion in the acute stage and total resolution without atrophy or cystic change. PMID- 20019109 TI - Quantification of diffusivities of the human cervical spinal cord using a 2D single-shot interleaved multisection inner volume diffusion-weighted echo-planar imaging technique. AB - BACKGROUND AND PURPOSE: DTI is a highly sensitive technique, which can detect pathology not otherwise noted with conventional imaging methods. This paper provides the atlas of reliable normative in vivo DTI parameters in the cervical spinal cord and its potential applications toward quantifying pathology. MATERIALS AND METHODS: In our study, we created a reference of normal diffusivities of the cervical spinal cord by using a 2D ss-IMIV-DWEPI technique from 14 healthy volunteers and compared parameters with those in 8 patients with CSM. The 2D ss-IMIV-DWEPI technique was applied in each subject to acquire diffusion-weighted images. FA, lambda( parallel), and lambda( perpendicular) were calculated. A reference of normal DTI indices from 12 regions of interest was created and compared with DTI indices of 8 patients. RESULTS: A map of reference diffusivity values was obtained from healthy controls. We found statistically significant differences in diffusivities between healthy volunteers and patients with CSM with different severities of disease, by using FA, lambda( parallel), and lambda( perpendicular) values. CONCLUSIONS: DTI using 2D ss-IMIV-DWPEI is sensitive to spinal cord pathology. This technique can be used to detect and quantify the degree of pathology within the cervical spinal cord from multiple disease states. PMID- 20019110 TI - Periscalene soft tissue: the new imaging hallmark in Erb palsy. AB - BACKGROUND AND PURPOSE: Pseudomeningocele is a well-known MR imaging finding in Erb palsy. Our aim was to evaluate the prevalence and imaging features of PST, a lesser known but, in our experience, more common finding in Erb palsy. MATERIALS AND METHODS: This retrospective study consisted of 37 subjects with Erb palsy from birth trauma. MR imaging examinations of the brachial plexus were evaluated independently by 2 neuroradiologists for the presence and signal-intensity characteristics/configuration of PST. Other findings included pseudomeningocele and posterior humeral head subluxation. The Blyth-Still-Casella procedure was used to construct an exact 95% CI for the percentage of patients seen to be positive for a specific imaging or anatomic feature by at least 1 observer and for the percentage positive according to both readers. RESULTS: The percentage of subjects found to have PST by both readers was 95% (35/37) (95% CI, 86.2%-99.9%). The PST was typically isointense to muscle on T1- and hyperintense on T2-weighted images. The most common morphology of the PST was a round or oval mass, though other morphologies were observed. Pseudomeningocele and posterior humeral head subluxation were reported in 40% (15/37) and 81% (30/37) of subjects, respectively. CONCLUSIONS: PST was the most common MR imaging finding in our series of subjects with Erb palsy and was observed more frequently than currently used imaging features. Although the morphology of PST is somewhat variable, several distinct patterns were observed, possibly reflecting different histologic compositions. PMID- 20019111 TI - A new canine carotid artery bifurcation aneurysm model for the evaluation of neurovascular devices. AB - BACKGROUND AND PURPOSE: Stents are increasingly used for coiling of difficult aneurysms, to reduce the risk of recurrences, or to modify blood flow. Currently available bifurcation aneurysm models are ill-suited to assess stent performance before clinical use. We designed a new wide-neck canine T-type bifurcation aneurysm model. Its potential value as a training tool as well as in the evaluation of new techniques or embolic agents was assessed. Our first task was to verify that recurrences occurred after satisfactory coiling. A second aim of this preliminary work was to assess if the new model could recreate the technical challenges involved in bifurcation aneurysms. MATERIALS AND METHODS: We introduce a new canine wide-neck bifurcation aneurysm model, created by using a vein pouch at the apex of an end-to-side anastomosis of the carotid arteries, with flow reversal in the proximal RCA by ligation of the innominate artery. Three aneurysms were treated with coil embolization, 10 were treated with stents (7 self-expandable, 3 balloon-expandable), and 3 were left untreated. Aneurysms were followed by duplex ultrasonography and angiography, and studied with macroscopic photography after euthanasia 11.8 +/- 3.9 months after surgery. RESULTS: All aneurysms remained patent at 9.0 +/- 3.6 months' follow-up. Coiling led to recurrences by 3 months in all 3 cases. Stent placement was technically difficult in all cases and did not lead to aneurysm thrombosis or neointimal closure of the aneurysm neck at 3 months. CONCLUSIONS: This model may be suitable for studying the effects of endovascular treatment on aneurysm and branch occlusion rates, for preclinical testing of stents and other intravascular devices, and for training students of endovascular technique. PMID- 20019112 TI - In vivo evaluation of the Phenox CRC mechanical thrombectomy device in a swine model of acute vessel occlusion. AB - BACKGROUND AND PURPOSE: Mechanical thrombectomy in ischemic stroke is of increasing interest as it is a promising strategy for fast and efficient recanalization. Several thrombectomy devices have been introduced to the armentarium of mechanical thrombectomy. Currently, new devices are under development and are continuously added to the neurointerventional tool box. Each device advocated so far has a different design and mechanical properties in terms of thrombus-device interaction. Therefore, a systematic evaluation under standardized conditions in vivo of these new devices is needed. The purpose of this study was to evaluate the efficiency, thrombus-device interaction, and potential complications of the novel Phenox CRC for distal mechanical thrombectomy in vivo. MATERIALS AND METHODS: The device was evaluated in an established animal model in the swine. Recanalization rate, thromboembolic events, vasospasm, and complications were assessed. Radiopaque thrombi (2 cm length) were used for the visualization of thrombus-device interaction during retrieval. The Phenox CRC (4 mm diameter) was assessed in 15 vessel occlusions. For every occlusion a maximum of 3 retrieval attempts were performed. RESULTS: Complete recanalization (TICI 3/TIMI 3) was achieved in 86.7% of vessel occlusions. In 66.7% (10/15), the first retrieval attempt was successful, and in 20% (3/15), the second attempt led to complete recanalization of the parent artery. In 2 cases (13.3%) thrombus retrieval was not successful (TICI 0/TIMI 0). In 1 case (6.7%) a minor embolic event occurred in a small side branch. No distal thromboembolic event was observed during the study. Thrombus-device interaction illustrated the entrapment of the thrombus by the microfilaments and the proximal cage of the device. No significant thrombus compression was observed. No vessel perforation, dissection, or fracture of the device occurred. CONCLUSIONS: In this small animal study, the Phenox CRC was a safe and effective device for mechanical thrombectomy. The unique design with a combination of microfilaments and proximal cage reduces thrombus compression with a consequently high recanalization and low complication rate. PMID- 20019113 TI - Penumbra system: a novel mechanical thrombectomy device for large-vessel occlusions in acute stroke. AB - BACKGROUND AND PURPOSE: Large IC artery occlusion is often resistant to recanalization. We present our initial experience with the PS. MATERIALS AND METHODS: Presenting with a severe acute ischemic stroke, the first 27 consecutive patients were considered for thromboaspiration therapy and retrospective data base analysis. All patients received standard thrombectomy treatment as monotherapy or in combination with thrombolysis or IC stent placement. The primary end point was revascularization of the target vessel to grade 2 or 3 on the TICI scale. Secondary end points were improvement of >4 points on the NIHSS score at discharge and favorable outcome, and improvement in overall mortality at 3 months and in sICH- and procedure-related adverse events. RESULTS: At baseline, the mean age was 66 +/- 14 years and the mean NIHSS score was 14 +/- 7. The anterior circulation was affected in 23 patients, and there were 4 basilar artery occlusions. Intracranial stent placement was performed in 4 patients. A recanalization to TICI 2 or 3 was achieved in 25 patients (93%). None of the patients developed sICH. At hospital discharge, 15 patients (56%) had an NIHSS improvement of >4 and 13 patients (48%) had an mRS score of <2 at 3 months. There was a significant correlation between complete vessel recanalization and favorable outcome. The all-cause mortality at 3 months was 11%. CONCLUSIONS: The PS showed a high potential for recanalization of acute thromboembolic occlusions of the large cerebral arteries. Complete recanalization was strongly correlated with good clinical outcome. PMID- 20019114 TI - Temporal trends in health-related quality of life among hemodialysis patients in the United States. AB - BACKGROUND AND OBJECTIVES: Health-related quality of life (HRQOL) is a measure of the well being of hemodialysis patients and an independent prognostic predictor. Our aim was to determine whether HRQOL among hemodialysis patients has changed over time. DESIGN, SETTING, PARTICIPANTS, & MEASUREMENTS: We retrospectively analyzed data collected by Dialysis Clinic, Inc. from adult patients starting hemodialysis between January 1, 1997 and May 31, 2006. The primary outcome was HRQOL assessed by Short Form 36, 6 to 18 months after and closest to the 1-year anniversary of starting hemodialysis. Secular trends were analyzed by linear regression for continuous variables and logistic regression for categorical ones. Year of starting dialysis was the predictor. A five-point difference on a 0 to 100 scale was considered clinically significant. RESULTS: Short Form 36 scores were available for 11,079 patients. Role physical, general health, vitality, social functioning, and physical component summary scores were unchanged among patients over the study period. Statistically significant (P < 0.05) but clinically insignificant changes were observed in physical functioning (-0.2 points/yr), bodily pain (+0.2 points/yr), mental health (+0.15 points/yr), and mental component summary scores (+0.13 points/yr). Only role emotional showed clinically significant improvement. Trends were unchanged after adjusting for age, gender, race, diabetes, hemoglobin, phosphorous, Kt/V, and albumin. CONCLUSIONS: Most HRQOL domains showed either no statistically significant change or statistically but not clinically significant change over almost a decade. These results suggest that, despite important developments in hemodialysis care since 1997, little progress was made in improving HRQOL of hemodialysis patients. PMID- 20019115 TI - Lack of association between dialysis modality and outcomes in atheroembolic renal disease. AB - BACKGROUND AND OBJECTIVES: Atheroembolic renal disease (AERD) can require dialytic support. Because anticoagulation may trigger atheroembolization, peritoneal dialysis may be preferred to hemodialysis. However, the effect of dialysis modality on renal and patient outcomes in AERD is unknown. DESIGN, SETTINGS, PARTICIPANTS, & MEASUREMENTS: A subcohort of 111 subjects who developed acute/subacute renal failure requiring dialysis was identified from a larger longitudinal study of AERD. The main exposure of interest was dialysis modality (peritoneal versus extracorporeal therapies). Logistic regression was used to study the probability of renal function recovery. Times from dialysis initiation to death were studied using Cox's regression. RESULTS: Eighty-six patients received hemodialysis and 25 received peritoneal dialysis. The probability of renal function recovery was similar by dialysis modality (25% among hemodialysis patients and 24% among peritoneal dialysis patients; P = 0.873). During follow up, 58 patients died, 14 among peritoneal patients and 44 among hemodialysis patients (P = 0.705). In multivariable analysis, gastrointestinal tract involvement and use of statins maintained an independent effect on the risk of patient death. CONCLUSIONS: This study does not support the notion that one dialysis modality is superior to the other. However, the observational nature of the data precludes any firm conclusions. PMID- 20019116 TI - Low dietary sodium intake increases the death risk in peritoneal dialysis. AB - BACKGROUND AND OBJECTIVES: To explore the correlation between dietary sodium intake and cardiovascular and overall mortality, and then determine whether this correlation can be explained by protein and energy intake paralleled with sodium intake in dialysis patients. DESIGN, SETTING, PARTICIPANTS, & MEASUREMENTS: This single-center retrospective cohort study enrolled 305 incident patients who started peritoneal dialysis in our unit from July 2002 to February 2007. All patients were followed until death or until being censored in February 2008. Demographic data were collected at baseline. Biochemical, dietary, and nutrition data were examined at baseline and thereafter at regular intervals to calculate the average values throughout the study. RESULTS: Participants with the highest average sodium intake were more likely to be younger, male, and overweight. Patients in the high tertile of average sodium intake had higher albumin, prealbumin, and lean body mass levels, and more nutrient intakes paralleling with sodium intake. Low average sodium intake independently predicted the increased risk for overall and cardiovascular death after adjusting for recognized confounders. Further adjustment for dietary protein, energy, and other nutrient intakes individually had minimal impact on the association between average sodium intake and overall death, with hazard ratios varying between 0.35 and 0.44, and cardiovascular death, with hazard ratios varying between 0.06 and 0.11. CONCLUSIONS: This study revealed that low dietary sodium intake independently predicts the high overall and cardiovascular mortality in dialysis patients. This correlation could not be entirely explained by deficient protein and energy intake. PMID- 20019117 TI - Agreement between central venous and arterial blood gas measurements in the intensive care unit. AB - BACKGROUND AND OBJECTIVES: Venous blood gas (VBG) analysis is a safer procedure than arterial blood gas (ABG) analysis and may be an alternative for determining acid-base status. The objective of this study was to examine the agreement between ABG and central VBG samples for all commonly used parameters in a medical intensive care unit (ICU) population. DESIGN, SETTING, PARTICIPANTS, & MEASUREMENTS: We performed a single-center, prospective trial to assess the agreement between arterial and central VBG measurements in a medical ICU. Adult patients who were admitted to the ICU and required both a central venous line and an arterial line were enrolled. When an ABG was performed, a central venous sample was obtained to examine the agreement among the pH, Pco(2), and bicarbonate. Data comparing central and peripheral VBG values were also obtained. RESULTS: The mean arterial minus venous difference for pH, Pco(2), and bicarbonate was 0.027, -3.8, and -0.80, respectively. Bland-Altman plots for agreement of pH, Pco(2), and bicarbonate showed 95% limits of agreement of -0.028 to 0.081, -12.3 to 4.8, and -4.0 to 2.4, respectively. Regression equations were derived to predict arterial values from venous values as follows: Arterial pH = 0.307 + 1.05 x venous pH, arterial Pco(2) = 0.805 + 0.936 x venous Pco(2), and arterial bicarbonate = 0.513 + 0.945 x venous bicarbonate. The mean central minus peripheral differences for pH, Pco(2), and bicarbonate were not clinically important. CONCLUSIONS: Peripheral or central venous pH, Pco(2), and bicarbonate can replace their arterial equivalents in many clinical contexts encountered in the ICU. PMID- 20019118 TI - The effect of measuring ambulatory blood pressure on nighttime sleep and daytime activity--implications for dipping. AB - BACKGROUND AND OBJECTIVES: Ambulatory blood pressure (BP) monitoring is commonly used to assess the circadian pattern of BP. Circadian BP pattern is influenced by physical activity and sleep cycle. The effect of BP monitoring itself on the level of physical activity and sleep remains unknown. If BP monitoring affects these parameters, then monitoring itself may influence the circadian BP pattern. DESIGN, SETTING, PARTICIPANTS, & MEASUREMENTS: To assess the effect of ambulatory BP monitoring on sleep duration, sleep efficiency, and daytime activity, we measured physical activity using wrist actigraphy in 103 veterans with chronic kidney disease. After 6 to 7 days of continuous activity monitoring, participants underwent ambulatory BP monitoring with simultaneous actigraphy. The above experiment was repeated after 1 mo. RESULTS: Among the top tertile of patients (most sleep), when wearing ambulatory BP patients spent less time in bed at night (-92 min, P < 0.0001), were less asleep during those hours (-98 min, P < 0.0001), and had reduced sleep efficiency (82% versus 77%, -5% P = 0.02). On the day of ambulatory BP monitoring, patients were more sedentary during waking hours (+27 minutes, P = 0.002). During ambulatory BP monitoring, waking after sleep onset more than median was associated with greater odds for nondipping (odds ratio 10.5, P = 0.008). CONCLUSIONS: Ambulatory BP monitoring is associated with disturbed sleep and reduced physical activity, characteristics that influence dipping. Ambulatory BP monitoring may itself induce nondipping and may thus mitigate the prognostic significance of the dipping phenomenon. PMID- 20019119 TI - Introduction to Focus on Fabry nephropathy: biomarkers, progression, and disease severity. PMID- 20019120 TI - A comparison of SF-36 and SF-12 composite scores and subsequent hospitalization and mortality risks in long-term dialysis patients. AB - BACKGROUND AND OBJECTIVES: The Short Form 12 (SF-12) has not been validated for long-term dialysis patients. The study compared physical and mental component summary (PCS/MCS) scores from the SF-36 with those from the embedded SF-12 in a national cohort of dialysis patients. DESIGN, SETTING, PARTICIPANTS, & MEASUREMENTS: All 44,395 patients who had scorable SF-36 and SF-12 from January 1, 2006, to December 31, 2006, and were treated at Fresenius Medical Care, North America facilities were included. Death and first hospitalization were followed for up to 1 year from the date of survey. Correlation and agreement were obtained between PCS-36 and PCS-12 and MCS-36 and MCS-12; then Cox models were constructed to compare associated hazard ratios (HRs) between them. RESULTS: Physical and mental dimensions both exhibited excellent intraclass correlation coefficients of 0.94. Each incremental point for both PCS-12 and PCS-36 was associated with a 2.4% lower adjusted HR of death and 0.4% decline in HR for first hospitalization (both P < 0.0001). Corresponding improvement in HR of death for each MCS point was 1.2% for MCS-12 and 1.3% for MCS-36, whereas both had similar 0.6% lower HR for hospitalization per point (all P < 0.0001). CONCLUSIONS: The use of the SF-12 alone or as part of a larger survey is valid in dialysis patients. Composite scores from the SF-12 and SF-36 have similar prognostic association with death and hospitalization risk. Prospective longitudinal studies of SF-12 surveys that consider responsiveness to specific clinical, situational, and interventional changes are needed in this population. PMID- 20019121 TI - Can the response to iron therapy be predicted in anemic nondialysis patients with chronic kidney disease? AB - BACKGROUND AND OBJECTIVES: Anemia is iron responsive in 30 to 50% of nondialysis patients with chronic kidney disease (CKD), but the utility of bone marrow iron stores and peripheral iron indices to predict the erythropoietic response is not settled. We investigated the accuracy of peripheral and central iron indices to predict the response to intravenous iron in nondialysis patients with CKD and anemia. DESIGN, SETTING, PARTICIPANTS, & MEASUREMENTS: A diagnostic study was conducted on 100 nondialysis patients who had CKD and anemia and were erythropoiesis-stimulating agent and iron naive. Bone marrow iron stores were evaluated by aspiration. Hemoglobin, transferrin saturation index (TSAT), and ferritin were measured at baseline and 1 month after 1000 mg of intravenous iron sucrose. Posttest predictive values for the erythropoietic response (> or =1-g/dl increase in hemoglobin) of peripheral and central iron indices were calculated. RESULTS: The erythropoietic response was noted in a higher proportion in bone marrow iron-deplete than in iron-replete patients (63 versus 30%). Peripheral iron indices had a moderate accuracy in predicting response. The positive (PPV) and negative predictive values (NPV) were 76 and 72% for a TSAT of 15% and 74 and 70% for a ferritin of 75 ng/ml, respectively. In the final logistic regression model, including TSAT and ferritin, the chances of a positive response increased by 7% for each 1% decrease in TSAT. CONCLUSIONS: Because an erythropoietic response is seen in half of patients and even one third of those with iron replete stores responded whereas peripheral indices had only a moderate utility in predicting response, the therapeutic trial to intravenous iron seems to be a useful tool in the management of anemia in nondialysis patients with CKD. PMID- 20019122 TI - Relationship between epoetin alfa dose and mortality: findings from a marginal structural model. AB - BACKGROUND AND OBJECTIVES: Observational studies relating epoetin alfa (EPO) dose and mortality frequently use analytic methods that do not control time-dependent confounding by indication (CBI). The relationship between EPO dose and 1-year mortality, adjusting for the effects of time-dependent CBI, was examined using a marginal structural model. DESIGN, SETTING, PARTICIPANTS, & MEASUREMENTS: This retrospective cohort study included 27,791 hemodialysis patients between July 2000 and June 2002. Patients were grouped at successive 2-wk intervals into a zero-dose category or four nonzero-dose categories. Ordinal regression was used to calculate inverse probability of treatment weights of patients receiving their own dose level given their covariate and treatment history. Three treatment models with an increasing number of treatment predictors were evaluated to assess the effect of model specification. A small number of excessively large patient weights were truncated. Relative hazards for higher-dose groups compared with the lowest nonzero-dose group varied by treatment model specification and by level of weight truncation. RESULTS: Results differed appreciably between the simplest treatment model, which incorporated only hemoglobin and EPO dosing history with 2% weight truncation (hazard ratio: 1.51; 95% confidence interval: 1.09, 1.89 for highest-dose patients), and the most comprehensive treatment model with 1% weight truncation (hazard ratio: 0.98; 95% confidence interval: 0.76, 1.74). CONCLUSIONS: There is appreciable CBI at higher EPO doses, and EPO dose was not associated with increased mortality in marginal structural model analyses that more completely addressed this confounding. PMID- 20019123 TI - A comparison of quality of life and travel-related factors between in-center and satellite-based hemodialysis patients. AB - BACKGROUND AND OBJECTIVES: Shorter travel times and distance to dialysis clinics have been associated with improved patient outcomes and a higher health-related quality of life (HRQOL). The objective of this study was to compare HRQOL between prevalent in-center and satellite dialysis patients, as well as compare travel related factors that contribute to HRQOL between in-center and satellite-based patients. DESIGN, SETTING, PARTICIPANTS, & MEASURES: The London Health Sciences Centre is a tertiary care center with in-center and regional satellite hemodialysis units. Patients who consented and completed a questionnaire (n = 202) were enrolled into a cross-sectional, cohort observational study. Patients were administered the Medical Outcomes Short-Form 36 (SF-36) and the Kidney Disease Health Related Quality of Life (KDHRQOL) tool and were asked questions relating to travel to dialysis clinics. RESULTS: Patients who underwent dialysis in the satellites had similar demographics, comorbidities, and laboratory parameters. Patients who underwent dialysis in satellite units reported a significantly superior score on the dialysis stress domain of the KDHRQOL questionnaire. There was no significant difference between in-center and satellite patients on the basis of the SF-36. Satellite patients also reported a significantly decreased cost of transportation, a significantly increased proportion who drive themselves to clinics, and significantly decreased travel time. CONCLUSIONS: Patients who underwent dialysis in satellite units demonstrated similar characteristics, comorbidities, surrogate outcomes, and most aspects of HRQOL. Travel time, cost, and receiving treatment in one's own community are important factors that may contribute to a trend toward higher reported HRQOL by patients in satellite dialysis units. PMID- 20019124 TI - The year in G protein-coupled receptor research. AB - I (R.P.M.) presented "The Year In G Protein-Coupled Receptor Research" at ENDO 2009. I first described the diversity of ligands and the five families into which the approximately 800 G protein-coupled receptors (GPCRs) are grouped, their basic structural architectures, their preeminent role in signaling, and the enormous scope for developing drugs targeted at GPCRs. I then spoke about some of the exciting breakthroughs in solving the atomic level structures of the active state of rhodopsin, beta(2)-adrenergic, beta(1)-adrenergic, and A(2A)-adenosine receptors. I also described studies on the structural changes accompanying the activation of the rhodopsin family of GPCRs. From these recent technical advances, we can anticipate that many more GPCR structures will emerge, which will afford us greater insight into their common and unique structural features and, particularly, the mechanisms underlying their activation. These insights will guide us in our understanding of how GPCRs operate, both in the normal and pathological situation. Although these crystal structures are highly informative, it is important to recognize that they represent static frozen conformations of a single GPCR state. New biophysical techniques are therefore being utilized to facilitate the dynamic monitoring of GPCR structural changes in relation to ligand activation. Solving of the crystal structures of GPCRs has also presented the real possibility of using the information of the ligand-binding pocket to allow in silico screening for novel small-molecule ligands. I then reviewed the concept of ligand-induced selective signaling of GPCRs, which is opening up new insights into more selective drug development. The assembly of GPCRs as homo- and heterooligomers and their phosphorylation and association with a vast array of trafficking and signal-modulating proteins are emerging as major mechanisms underlying the functioning of GPCRs. Differential expression and recruitment of these proteins provide a mechanism for subtle physiological regulation of cellular activity. Finally, I mentioned some of the GPCRs that have lately come to the fore as novel regulators in endocrinology. These included fatty acid specific GPCRs expressed in pancreatic beta-cells and novel neuroendocrine GPCRs regulating reproduction. PMID- 20019125 TI - Introduction to the year in basic science series...2009. AB - This year represents the second, of what we expect to be an annual series, of articles based on The Endocrine Society annual meeting presentations that highlight recent advances in vibrant basic science areas in endocrinology. For ENDO 09, two general areas with broad appeal and significance to our members were chosen: neuroendocrinology and G protein-coupled receptors. The invited participants were charged with presenting and discussing important papers that were published approximately during the year leading up to the most recent annual meetings (June 2009) and to put them into broad perspective for the greater endocrine community. Two distinguished members, Jeffrey Blaustein and Robert Millar, continued on last year's successful features by synthesizing the top findings in their fields, and these articles are based on their annual meeting presentations. Interestingly, there were several points of intersection in these topics and chosen papers, as advances in the neuroendocrinology of reproduction have been coupled to identification and/or characterization of additional novel G protein-coupled receptors. In both presentations, fundamental basic science findings deriving from structural studies and signaling pathways are linked to broad endocrine physiology issues and to potential use in clinical treatment and therapeutics. PMID- 20019126 TI - The Year in Neuroendocrinology. AB - At the "Year In Neuroendocrinology" session during the 2009 meeting of The Endocrine Society, I highlighted recent progress in three main areas of neuroendocrinology: neural mechanisms of action of estradiol, GnRH regulation, and epigenetics. In the area of neural mechanisms of action of estradiol, we have seen the list of estrogen receptors continue to expand and that neurosteroid synthesis is rapidly regulated by the social environment; that brain sexual differentiation can occur via the action of estradiol in upstream neurons without the need of estrogen receptors in affected neurons; and that a particular xenoestrogen can block the effects of estradiol and testosterone in brain synapses. In the area of GnRH regulation, kisspeptin continues to be a major player in reproductive endocrinology; neurokinin B should be added to the growing list of critical peptides involved in puberty and reproduction; and RFamide related peptides have a direct role in regulation of gonadotropin-releasing neurons as a gonadotropin-inhibiting hormone. Finally, in the epigenetics field, we learned that the same principles of importance of parental care in epigenetic regulation in hippocampal glucocorticoid receptors in rats applies to humans and may explain some long-term effects of childhood abuse on the hypothalamic pituitary-adrenal axis. It is difficult, if not impossible, to predict which findings will have an enduring impact on the field. It will be interesting to look back, 10 yr from now, to see whether the papers that were chosen were in fact well cited and whether they were influential in driving additional research in neuroendocrinology. PMID- 20019128 TI - Cigarette smoking and mortality risk in people with schizophrenia. AB - This study examined effects of cigarette smoking on mortality risk in 1213 persons aged 19-69 years with schizophrenia-related psychotic disorders admitted to State of Maryland Hospitals between 1994 and 2000. Inpatient medical records from 7 hospitals were reviewed to obtain demographic information, diagnosis, medication use, as well as smoking and other substance use. Social Security Death Index data were used to identify deaths in the study group between 1994 and 2004. Death records were reviewed to obtain manner of death and underlying disorders. Of the 1213, 55% were smokers and 71% abused substances. There was an age * smoking interaction (chi(2) = 14.6, df = 1, P = .0001) for mortality, with estimated hazard ratios (HRs) for smokers vs nonsmokers of 2.1 among 35- to 54 year olds and HR of 0.7 among those aged 55-69 years. Five- and 10-year mortality rates for smokers aged 35-54 years were 7.0% and 14.2%, compared with 3.3% and 10.0% for nonsmokers, respectively (chi(2) = 5.53, df = 1, P = .019). Cardiac causes were identified in 43% of deaths in smokers but only 19% of deaths in nonsmokers (P < .006). For those aged 35-54 years, the odds of cardiac related death was increased by 12 fold in smokers relative to nonsmokers (HR = 12.4, chi(2) = 12.0, df = 1, P = .0005). Among people aged 35-54 years, those smoking greater than one pack daily have a significantly increased total mortality risk (HR = 2.7) vs nonsmokers. Cigarette smoking, particularly in people aged 35-54 years, contributes to an increased risk of death. Greater smoking severity significantly increases this risk. Smoking cessation in people with schizophrenia deserves significant attention. PMID- 20019129 TI - Serum glucocorticoids and adiponectin associate with insulin resistance in children born small for gestational age. AB - OBJECTIVES: Altered glucocorticoid activity is one possible mechanism linking fetal growth restriction with later insulin resistance (IR) and type 2 diabetes. We aimed to investigate whether serum glucocorticoid parameters are related to IR in children born small for gestational age (SGA). DESIGN: A total of 110 children (55 age- and gender-matched pairs born SGA or appropriate for gestational age (AGA) in a case-control setting) were studied at the mean age of 12.2 (s.d. 0.2) years. METHODS: Serum cortisol, corticosteroid-binding globulin (CBG), free cortisol index (FCI=cortisol/CBG), and glucocorticoid bioactivity (GBA, transactivation assay) were analyzed and related to serum adiponectin and insulin like growth factor-binding protein 1 (IGFBP1) concentrations and homeostasis model assessment for IR (HOMA-IR) and QUICKI indices. RESULTS: In the pooled study population, GBA correlated well with cortisol and FCI (r=0.681 and 0.586 respectively; P<0.001 for both). Serum cortisol, CBG, FCI, GBA, HOMA-IR, or QUICKI did not differ between the SGA and AGA subjects, but the SGA children had lower body mass index (P=0.005) and waist circumference (WC) (P=0.001). The mean GBA in the highest GBA quartile was higher among the SGA subjects than among the AGA subjects (138.6 vs 96.4 nmol/l cortisol equivalents, P<0.001). In the SGA children, GBA correlated positively with HOMA-IR (r=0.522, P<0.001) and inversely with adiponectin (r=-0.278, P=0.042) (WC/height ratio adjustments), and in logistic regression analysis, higher GBA (odds ratio (OR) 1.3; P=0.013), lower adiponectin (OR 1.4; P=0.038), and lower IGFBP1 (OR 1.9; P=0.010) associated independently with higher HOMA-IR. CONCLUSIONS: These findings suggest that increased glucocorticoid activity and low serum adiponectin concentrations associate with IR in SGA children. PMID- 20019130 TI - Children with Prader-Willi syndrome exhibit more evident meal-induced responses in plasma ghrelin and peptide YY levels than obese and lean children. AB - BACKGROUND AND AIMS: Ghrelin is an orexigenic 28-amino acid peptide produced by the stomach. Circulating ghrelin levels rise shortly before and fall shortly after every meal. Peptide YY (PYY), an anorexigenic 36-amino acid peptide, is secreted primarily from the intestinal mucosa of the ileum and large intestine. Plasma PYY levels begin to rise within 15 min after starting to eat and plateau within approximately 90 min, remaining elevated for up to 6 h. Recently, some studies have tried to evaluate the potential role of ghrelin and PYY in the hyperphagia of patients with Prader-Willi syndrome (PWS). While hyperghrelinemia is well characterized in PWS, conflicting results have been reported for PYY. The aim of the study was to investigate ghrelin and PYY responses to a standard liquid high-fat meal in children with PWS. PATIENTS AND METHODS: Circulating levels of total ghrelin and PYY levels were assayed by RIA after overnight fasting and 45, 60, 90, and 180 min following a standard meal (Ensure 6 ml/kg) in 16 patients with PWS (11 boys and five girls, aged 4.6-10.7 years, including ten receiving 0.02 mg/kg per day rhGH for 2-18 months; body mass index (BMI) z-score: 0.6+/-0.2 and 1.6+/-0.5 for children treated or not treated with rhGH respectively), ten obese (eight boys and two girls, aged 9.2-15.6 years; BMI z score: 2.4+/-0.2, i.e. BMI >97th centile for chronological age and sex) subjects, and 16 normal-weight controls (five boys and 11 girls, aged 5.8-17.3 years; BMI z score: 0.6+/-0.2). RESULTS: PWS children showed higher fasting levels of ghrelin than obese and lean controls. Postprandial ghrelin drop was more pronounced in PWS than in the other study groups. No significant difference on fasting levels of PYY was found among groups. PWS showed a higher postprandial PYY rise than obese and lean controls. PWS patients treated and not treated with GH showed similar fasting and postprandial levels of ghrelin and PYY. Fasting PYY levels correlated negatively (P<0.05; r=-0.68) with those of ghrelin only in PWS. CONCLUSIONS: The results of this study confirm fasting hyperghrelinemia in PWS. Since in PWS adults an impaired postprandial suppression of plasma ghrelin was previously reported to be associated with a blunted postprandial PYY response, the finding of a meal-induced decrease and increase in ghrelin and PYY levels respectively in PWS children would imply that the regulation of appetite/satiety of these peptides is operative during childhood, and it progressively deteriorates and vanishes in adulthood when hyperphagia and obesity worsen. PMID- 20019131 TI - Primary breast carcinoma: association of mammographic calcifications with osteopontin expression. AB - PURPOSE: To evaluate the osteopontin (OPN) protein expression levels in breast carcinomas to determine if they correlate with mammographic appearances such as calcifications. MATERIALS AND METHODS: This retrospective study was institutional review board approved. Informed consent was obtained from patients. Clinical history, histopathologic findings, mammographic features, and OPN expression as determined with immunohistochemistry results were evaluated in 141 women with breast cancer. The median age of patients was 53 years (range, 29-82 years). Mammographic features and clinicopathologic characteristics were correlated with tumor OPN expression. chi(2) And Fisher exact tests were used to evaluate the association of OPN expression with mammographic and clinicopathologic features. RESULTS: Calcifications on mammograms (P = .012), spiculated margins of mass on mammograms (P = .02), "triple-negative" (ie, cancer that is estrogen receptor, progesterone receptor, and human epidermal growth factor receptor negative) phenotype (P = .02), and lymph node metastasis (P < .0001) were significantly associated with OPN status. In contrast to OPN-negative tumors, OPN-positive tumors were more likely to have spiculated margins (57.6% vs 9.2%), to be associated with calcifications (54.3% vs 30.6%), to be a triple-negative phenotype (26% vs 8.1%), and to have axillary lymph node metastasis (81.5% vs 38.8%). Most calcifications were of pleomorphic morphology (60.4% vs 11.8%, P = .046). CONCLUSION: OPN could play a role in the formation of calcifications that often are associated with breast cancer. PMID- 20019127 TI - Estrogen therapy and cognition: a review of the cholinergic hypothesis. AB - The pros and cons of estrogen therapy for use in postmenopausal women continue to be a major topic of debate in women's health. Much of this debate focuses on the potential benefits vs. harm of estrogen therapy on the brain and the risks for cognitive impairment associated with aging and Alzheimer's disease. Many animal and human studies suggest that estrogens can have significant beneficial effects on brain aging and cognition and reduce the risk of Alzheimer's-related dementia; however, others disagree. Important discoveries have been made, and hypotheses have emerged that may explain some of the inconsistencies. This review focuses on the cholinergic hypothesis, specifically on evidence that beneficial effects of estrogens on brain aging and cognition are related to interactions with cholinergic projections emanating from the basal forebrain. These cholinergic projections play an important role in learning and attentional processes, and their function is known to decline with advanced age and in association with Alzheimer's disease. Evidence suggests that many of the effects of estrogens on neuronal plasticity and function and cognitive performance are related to or rely upon interactions with these cholinergic projections; however, studies also suggest that the effectiveness of estrogen therapy decreases with age and time after loss of ovarian function. We propose a model in which deficits in basal forebrain cholinergic function contribute to age-related changes in the response to estrogen therapy. Based on this model, we propose that cholinergic-enhancing drugs, used in combination with an appropriate estrogen-containing drug regimen, may be a viable therapeutic strategy for use in older postmenopausal women with early evidence of mild cognitive decline. PMID- 20019132 TI - Crush thoracic trauma in the massive Sichuan earthquake: evaluation with multidetector CT of 215 cases. AB - PURPOSE: To determine the multidetector computed tomographic (CT) features of crush thoracic traumas resulting from the massive Sichuan earthquake. MATERIALS AND METHODS: This study was approved by the ethics committee of the medical school, and informed consent was waived. A retrospective review was undertaken of 215 multidetector chest CT scans of 112 male and 103 female patients who sustained crush thoracic injuries in the Sichuan earthquake at 2:28 pm Beijing time, May 12, 2008, and were rescued in the authors' hospital. Multidetector chest CT studies were performed between May 12, 2008, and June 7, 2008. The authors looked for injuries to the thoracic cage, pulmonary parenchyma, and pleura. RESULTS: One hundred forty-three patients (66.5%; 95% confidence interval [CI]: 60.2%, 72.8%) had at least one rib fracture; the mean number of rib fractures per patient was 6 [corrected]. Forty-five of these patients (31.5% of 143 patients; 95% CI: 23.9%, 39.1%) had flail chest, with a total of 288 ribs fractured. There were 46 patients (21.4%; 95% CI: 15.9%, 26.9%) with at least one vertebral fracture. There were 77 vertebral fractures total; 36 of these fractures were in T1 through T10. Twelve patients (5.6%; 95% CI: 2.5%, 8.7%) had sternal fractures, and 48 patients (22.3%; 95% CI: 16.7%, 27.9%) had either scapular or clavicular fractures. There were 117 patients (54.4%; 95% CI: 47.7%, 61.1%) with pulmonary parenchymal injuries and 146 (67.9%; 95% CI: 61.7%, 74.1%) with pleural injuries. CONCLUSION: Crush thoracic trauma resulting from the massive Sichuan earthquake was a life-threatening injury; this type of injury has the potential for multiple fractures and pulmonary parenchymal injuries. PMID- 20019133 TI - Evaluation of anatomic valve opening and leaflet morphology in aortic valve bioprosthesis by using multidetector CT: comparison with transthoracic echocardiography. AB - PURPOSE: To prospectively determine whether cardiac-gated multidetector computed tomography (CT) allows visualization of aortic valve leaflets after bioprosthetic aortic valve replacement (AVR), to provide an accurate method for measuring the aortic valve opening, and to provide morphologic and functional information regarding the mechanism underlying poor function of the bioprosthetic valve. MATERIALS AND METHODS: The institutional review board approved the study protocol; informed consent was given. Fifty-four patients (27 men; mean age, 75 years + or - 8 [standard deviation]) with bioprosthetic AVR implanted 2 years + or - 3 earlier underwent 64-section CT and transthoracic echocardiography (TTE). Two blinded observers manually planimetered the aortic valve area (AVA) by using a computer workstation on end-systolic short-axis CT images and measured opening angles (OAs) between the bioprosthesis annulus base and the free margin on long axis images. These measurements were compared with those of the effective orifice area (EOA) of the valve at Doppler continuity-equation TTE by using regression and Bland-Altman methods. Morphology and mobility of leaflets in normally functioning (EOA indexed to body surface area [EOA(i)] > 0.65 cm(2)/m(2)) and dysfunctional (EOA(i)< 0.65 cm(2)/m(2)) AVRs were compared. RESULTS: AVA at CT correlated highly to EOA at TTE (r = 0.93, P < .001) but was significantly larger (1.2 cm(2) + or - 0.4 vs 1.1 cm(2) + or - 0.3, P < .001) than EOA at TTE. In dysfunctional bioprostheses (n = 34), CT results showed a variety of morphologic abnormalities, such as leaflet thickening (n = 9), presumed thrombotic material (n = 6), and leaflet calcification (n = 1). Multidetector CT results demonstrated restriction of leaflet motion indicated by lower OA (64 degrees + or - 5 vs 79 degrees + or - 3, P < .0001) in dysfunctional AVRs than in normally functioning AVRs (n = 11). CONCLUSION: Sixty-four-section CT can help accurately measure AVA in bioprosthetic AVR compared with EOA at TTE. It can also show morphologic abnormalities and reduced leaflet motion in a dysfunctional bioprosthesis, thereby potentially unraveling the mechanism of dysfunction. PMID- 20019136 TI - Value of US correlation of a thyroid nodule with initially benign cytologic results. AB - PURPOSE: To investigate the value of ultrasonographic (US) features in thyroid nodules with initially benign cytologic results. MATERIALS AND METHODS: The institutional review board approved this retrospective study and required neither patient approval nor informed consent for the review of images and records. From October 2003 to February 2006, 6118 focal thyroid nodules in 6025 consecutive patients underwent US-guided fine-needle aspiration biopsy (FNAB). This study included 1343 nodules 1 cm or larger in 1302 patients that were diagnosed as benign at initial cytologic evaluation and underwent pathologic or follow-up study. We compared the risk of malignancy according to US findings and calculated the likelihoods of different subgroups having benign nodules. RESULTS: In total, 26 (1.9%) malignant and 1317 (98.1%) benign nodules were found according to reference standards. If initial cytologic results showed benign thyroid nodules, the likelihood of the nodule actually being benign was 98.1%. When a thyroid nodule had benign results at both initial and repeat FNAB, the likelihood increased to 100%. The likelihood of having a benign thyroid nodule with suspicious US features was lower (79.6%) than having a benign thyroid nodule with negative US features (99.4%, P<.001). In the nodule with benign features at initial US, the risk of malignancy for a thyroid nodule with an increase in size at follow-up US was slightly higher (1.4%) than that of a thyroid nodule with no interval change or decrease in size, but it was not significantly different (0.5%, P = .354). CONCLUSION: Repeat FNAB should be performed for thyroid nodules that have suspicious US features, even if the initial cytologic results indicate that it is a benign lesion. PMID- 20019135 TI - Multidetector CT angiography in the evaluation of acute blunt head and neck trauma: a proposed acute craniocervical trauma scoring system. AB - PURPOSE: To determine the diagnostic yield of multidetector computed tomographic (CT) angiography in the evaluation of patients presenting to the emergency department with acute blunt head and neck trauma to assess for arterial injury and to propose a practical scoring system for the identification of patients at highest risk of arterial injury. MATERIALS AND METHODS: With institutional review board approval, Health Insurance Portability and Accountability Act compliance, and waived informed consent, a retrospective study was conducted of 830 consecutive patients who presented to the emergency department with acute blunt head and neck trauma over 9 years and were evaluated with multidetector CT angiography. Unenhanced CT scans and CT angiograms were reviewed for the presence of cervical interfacetal subluxations and/or dislocations, fractures, intracranial hemorrhage, and arterial injury. Medical records were reviewed for mechanism of injury (MOI). Multivariate logistic regression analysis was performed to identify independent predictors of an increased risk of arterial injury. RESULTS: Multidetector CT angiographic results showed injury to 118 arterial structures in 106 (12.8%) patients. Multivariate logistic regression analysis showed that the presence of cervical interfacetal subluxation/dislocations (44.4%; odds ratio [OR], 6.3; P < .0001), fracture lines reaching an arterial structure (22.1%; OR, 4.4; P < .0001), and high-impact MOIs (16.5%; OR, 3.1; P < .0001) were independent predictors of an increased risk of arterial injury and were used to construct a scoring system. Patients with scores of 2 and 3 (21.9% and 52.2%, respectively) were at highest risk of arterial injury. CONCLUSION: The proposed acute craniocervical trauma scoring system could be used as a guide to select blunt trauma patients for multidetector CT angiographic evaluation. Future validation of this scoring system is necessary. PMID- 20019137 TI - Body size indexes for optimizing iodine dose for aortic and hepatic enhancement at multidetector CT: comparison of total body weight, lean body weight, and blood volume. AB - PURPOSE: To evaluate and compare total body weight (TBW), lean body weight (LBW), and estimated blood volume (BV) for the adjustment of the iodine dose required for contrast material-enhanced multidetector computed tomography (CT) of the aorta and liver. MATERIALS AND METHODS: Institutional review committee approval and written informed consent were obtained. One hundred twenty patients (54 men, 66 women; mean age, 64.1 years; range, 19-88 years) who underwent multidetector CT of the upper abdomen were randomized into three groups of 40 patients each: (a) TBW group (0.6 g of iodine per kilogram of TBW), (b) LBW group (0.821 g of iodine per kilogram of LBW), and (c) BV group (men, 8.6 g of iodine per liter of BV; women, 9.9 g of iodine per liter of BV). Change in CT number between unenhanced and contrast-enhanced images per gram of iodine and maximum hepatic enhancement (MHE) adjusted for iodine dose were examined for correlation with TBW, LBW, and BV by using linear regression analysis. RESULTS: In the portal venous phase, correlation coefficients for the correlation of change in CT number per gram of iodine with TBW for the aorta and liver were -0.71 and -0.79, respectively, in the TBW group; -0.80 and -0.86, respectively, in the LBW group; and -0.68 and -0.66, respectively, in the BV group. In the liver, they were marginally higher in the LBW group than in the BV group (P = .03). Adjusted MHE remained constant at 77.9 HU +/- 10.2 (standard deviation) in the LBW group with respect to TBW, but it increased in the TBW (r = 0.80, P < .001) and BV (r = 0.70, P < .001) groups as TBW increased. CONCLUSION: When LBW, rather than TBW or BV, is used, the iodine dose required to achieve consistent hepatic enhancement may be estimated more precisely and with reduced patient-to-patient variability. PMID- 20019138 TI - Accelerated two- and three-dimensional cine MR imaging of the heart by using a 32 channel coil. AB - PURPOSE: To compare accelerated real-time two-dimensional (2D) and segmented three-dimensional (3D) cine steady-state free precession magnetic resonance (MR) imaging techniques by using a 32-channel coil with a conventional 2D cine imaging approach for imaging the heart and to evaluate any difference caused by free breathing and breath holding for real-time imaging. MATERIALS AND METHODS: In this institutional review board-approved HIPAA-compliant study, 10 healthy volunteers and 22 consecutive patients who were suspected of having or were known to have heart disease underwent cardiac MR imaging by using a 32-channel coil. A conventional multisection 2D real-time cine sequence was used as the reference standard, and three additional accelerated cine sequences were implemented. Volumetric parameters, including ejection fraction (EF), end-diastolic volume (EDV), end-systolic volume (ESV), stroke volume(SV), and myocardial mass, were derived. Wall motion and image quality were assessed by two radiologists. In addition, image time was registered. An additional set of images was acquired by using real-time sequences with free breathing, and quantitative measurements were compared with measurements on images obtained with breath holding. For quantitative analysis, repeated-measures analysis of variance, paired t test, and Bland-Altman analysis were used; for qualitative analysis, nonparametric Wilcoxon signed-rank test was used. RESULTS: All volumetric measurements were significantly correlated with those of the standard sequence (r > 0.80, P < .01). No significant difference among protocols was observed in terms of mean levels for EF or ESV (P > .05). However, a significant difference was indicated for EDV and SV (P < .01).The accelerated protocols had significantly shorter image times (P < .001). Wall motion scores were concordant with the standard sequence in 43 44 (93%-96%) segments for the accelerated protocols, with a strong interreader agreement (intraclass correlation coefficient, > or =0.93). No significant difference was identified between real-time protocols with free breathing and those with breath holding for measurement of volumetric parameters. CONCLUSION: Accelerated real-time 2D and segmented 3D cine techniques are comparable to the standard clinical protocol in assessment of left ventricular global and regional parameters in substantially shorter image times. PMID- 20019139 TI - Locally advanced rectal carcinoma treated with preoperative chemotherapy and radiation therapy: preliminary analysis of diffusion-weighted MR imaging for early detection of tumor histopathologic downstaging. AB - PURPOSE: To determine whether changes in apparent diffusion coefficients (ADCs) of rectal carcinoma obtained 1 week after the beginning of chemotherapy and radiation therapy (CRT) correlate with tumor histopathologic downstaging after preoperative CRT. MATERIALS AND METHODS: This prospective study was approved by an institutional review board; informed consent was obtained from all patients. Thirty-seven patients (mean age, 54.7 years; 13 women, 24 men) with primary rectal carcinoma who were undergoing preoperative CRT were recruited for the study. Diffusion-weighted (DW) magnetic resonance (MR) imaging was performed with a 1.5-T MR imager in all patients before therapy, at the end of the 1st and 2nd week of therapy, and before surgery. Tumor ADCs were calculated. Linear mixed effects modeling was applied to analyze change in ADCs and volumes following treatment. RESULTS: Patients were assigned to the tumor downstaged group (n = 17) or the tumor nondownstaged group (n = 20) on the basis of histopathologic examination results following surgery. Before CRT, the mean tumor ADC in the downstaged group was lower than that in the nondownstaged group (1.07 x 10(-3) mm(2)/sec +/- 0.13 [standard deviation] vs 1.19 x 10(-3) mm(2)/sec +/- 0.15, F = 6.91, P = .013). At the end of the 1st week of CRT, the mean tumor ADC increased significantly from 1.07 x 10(-3) mm(2)/sec +/- 0.13 to 1.32 x 10(-3) mm(2)/sec +/ 0.16 (F = 37.63, P <.001) in the downstaged group, but there was no significant ADC increase in the nondownstaged group (F = 1.18, P = .291). The mean percentage of tumor ADC change in the downstaged group was significantly higher than that in the nondownstaged group at each time point (F = 18.39, P < .001). CONCLUSION: Early increase of mean tumor ADC and low pretherapy mean ADC in rectal carcinoma correlate with good response to CRT. DW MR imaging is a promising noninvasive technique for helping predict and monitor early therapeutic response in patients with rectal carcinoma who are undergoing CRT. PMID- 20019140 TI - Clinically isolated syndrome suggestive of multiple sclerosis: voxelwise regional investigation of white and gray matter. AB - PURPOSE: To quantify white matter (WM) and gray matter (GM) damage in patients who presented with clinically isolated syndrome (CIS), which is suggestive of multiple sclerosis (MS), by combining volume-based morphometry (VBM) and tract based spatial statistics (TBSS). MATERIALS AND METHODS: This prospective HIPAA compliant study was approved by the institutional review board. Written informed consent was obtained from all participants. In this study, 34 consecutive patients (21 women, 13 men; mean age, 31.7 years +/- 7.7 [standard deviation]) who presented with CIS were recruited. The magnetic resonance (MR) examination included dual-echo fast spin-echo, three-dimensional T1, and diffusion-tensor imaging. Sixteen matched healthy volunteers served as control subjects. T2 lesion volumes were assessed with a semiautomatic technique. VBM and TBSS were used for the GM and WM analyses, respectively, to compare regional GM volumes and fractional anisotropy (FA) values in the two groups. RESULTS: TBSS analysis revealed a pattern of diffuse FA reductions in patients with CIS at the cluster level (P < .05). Regions of decreased FA involved most of the WM pathways, including the corticospinal tracts, corpus callosum, and superior and inferior longitudinal fasciculi. There were no significant differences between the two groups in terms of global GM, WM, or cerebrospinal fluid volume or in terms of regional GM volume. CONCLUSION: Diffuse WM damage not accompanied by any change in GM or WM volume is observed in patients with CIS. This suggests that WM involvement plays a relevant role in the early phases of MS. Subsequently detected GM damage may be secondary to WM alterations. PMID- 20019141 TI - Patellar cartilage: T2 values and morphologic abnormalities at 3.0-T MR imaging in relation to physical activity in asymptomatic subjects from the osteoarthritis initiative. AB - PURPOSE: To study the interrelationship between patella cartilage T2 relaxation time, other knee abnormalities, and physical activity levels in asymptomatic subjects from the Osteoarthritis Initiative (OAI) incidence cohort. MATERIALS AND METHODS: The study had institutional review board approval and was HIPAA compliant. One hundred twenty subjects from the OAI without knee pain (age, 45-55 years) and with risk factors for knee osteoarthritis (OA) were studied by using knee radiographs, 3.0-T knee magnetic resonance (MR) images (including intermediate-weighted fast spin-echo and T2 mapping sequences), and the Physical Activity Scale for the Elderly. MR images of the right knee were assessed by two musculoskeletal radiologists for the presence and grade of abnormalities. Segmentation of the patella cartilage was performed, and T2 maps were generated. Statistical significance was determined by using analysis of variance, chi(2) analysis, correlation coefficient tests, the Cohen kappa, and a multiple linear regression model. RESULTS: Cartilage lesions were found in 95 (79.0%) of 120 knees, and meniscal lesions were found in 54 (45%) of 120 knees. A significant correlation between patella cartilage T2 values and the severity and grade of cartilage (P = .0025) and meniscus (P = .0067) lesions was demonstrated. Subjects with high activity levels had significantly higher prevalence and grade of abnormalities and higher T2 values (48.7 msec +/-4.35 vs 45.8 msec +/-3.93; P < .001) than did subjects with low activity levels. CONCLUSION: Middle-aged asymptomatic individuals with risk factors for knee OA had a high prevalence of cartilage and meniscus knee lesions. Physically active individuals had more knee abnormalities and higher patellar T2 values. Additional studies will be needed to determine causality. PMID- 20019142 TI - Evolution of radiation-induced brain injury: MR imaging-based study. AB - PURPOSE: To evaluate the temporal lobes in patients previously treated for nasopharyngeal carcinoma to provide a better understanding of delayed radiation induced injury in the brain unaffected by the underlying tumor. MATERIALS AND METHODS: Retrospective analysis of the patient data was approved by the local ethics committee. Informed consent was waived. Magnetic resonance (MR) imaging results in patients with temporal lobe injury (TLI) after receiving radiation for nasopharyngeal carcinoma were analyzed. The appearance and change over time of white matter lesions (WMLs), contrast material-enhanced lesions, and cysts were assessed. The Mann-Whitney U test was used to compare interval time, and the chi(2) and Fisher exact tests were used to compare the pattern of TLI changes. RESULTS: The study group was 124 patients (95 men, 29 women; mean age, 51.4 years) with 192 injured temporal lobes; 62 of these patients with 103 injured temporal lobes underwent follow-up MR imaging at least once (range, one to five examinations). A total of 332 injured temporal lobes were revealed. WMLs, contrast-enhanced lesions, and cysts were present on 332 (100%), 274 (82.5%), and 42 (12.7%) studies, respectively. All contrast-enhanced lesions more than 2 cm in size showed necrosis, and those 3 cm or greater formed a rim-enhanced necrotic mass. WMLs were the only lesion to occur alone, contrast-enhanced lesions were always accompanied by WMLs, and cysts were always accompanied by WMLs and contrast-enhanced lesions. Detection of cysts was significantly later than detection of WMLs and contrast-enhanced lesions (P <.01). Regression or resolution was found in 27 (28%) of 96 WMLs, 37 (39%) of 94 contrast-enhanced lesions, and one (7%) of 15 cysts. CONCLUSION: TLI from radiation is not always an irreversible and progressive process but is one that can regress or resolve at MR imaging. In the evolution of radiation injury, WMLs are seen first and are followed by contrast-enhanced lesions, which have an increasing tendency to become necrotic with increasing size. Cysts are the least frequent manifestation and arise in the late stage of TLI. PMID- 20019143 TI - A SNP discovery method to assess variant allele probability from next-generation resequencing data. AB - Accurate identification of genetic variants from next-generation sequencing (NGS) data is essential for immediate large-scale genomic endeavors such as the 1000 Genomes Project, and is crucial for further genetic analysis based on the discoveries. The key challenge in single nucleotide polymorphism (SNP) discovery is to distinguish true individual variants (occurring at a low frequency) from sequencing errors (often occurring at frequencies orders of magnitude higher). Therefore, knowledge of the error probabilities of base calls is essential. We have developed Atlas-SNP2, a computational tool that detects and accounts for systematic sequencing errors caused by context-related variables in a logistic regression model learned from training data sets. Subsequently, it estimates the posterior error probability for each substitution through a Bayesian formula that integrates prior knowledge of the overall sequencing error probability and the estimated SNP rate with the results from the logistic regression model for the given substitutions. The estimated posterior SNP probability can be used to distinguish true SNPs from sequencing errors. Validation results show that Atlas SNP2 achieves a false-positive rate of lower than 10%, with an approximately 5% or lower false-negative rate. PMID- 20019144 TI - De novo assembly of human genomes with massively parallel short read sequencing. AB - Next-generation massively parallel DNA sequencing technologies provide ultrahigh throughput at a substantially lower unit data cost; however, the data are very short read length sequences, making de novo assembly extremely challenging. Here, we describe a novel method for de novo assembly of large genomes from short read sequences. We successfully assembled both the Asian and African human genome sequences, achieving an N50 contig size of 7.4 and 5.9 kilobases (kb) and scaffold of 446.3 and 61.9 kb, respectively. The development of this de novo short read assembly method creates new opportunities for building reference sequences and carrying out accurate analyses of unexplored genomes in a cost effective way. PMID- 20019145 TI - Alterations in frontal lobe tracts and corpus callosum in young children with autism spectrum disorder. AB - Major frontal lobe tracts and corpus callosum (CC) were investigated in 32 children with autism spectrum disorder (ASD, mean age: 5 years), 12 nonautistic developmentally impaired children (DI, mean age: 4.6 years), and 16 typically developing children (TD, mean age: 5.5 years) using diffusion tensor imaging tractography and tract-based spatial statistics. Various diffusion and geometric properties were calculated for uncinate fasciculus (UF), inferior fronto occipital fasciculus (IFO), arcuate fasciculus (AF), cingulum (Cg), CC, and corticospinal tract. Fractional anisotropy was lower in the right UF, right Cg and CC in ASD and DI children; in right AF in ASD children; and in bilateral IFO in DI children, compared with TD children. Apparent diffusion coefficient was increased in right AF in both ASD and DI children. The ASD group showed shorter length of left UF and increased length, volume, and density of right UF; increased length and density of CC; and higher density of left Cg, compared with the TD group. Compared with DI group, ASD group had increased length, volume, and density of right UF; higher volume of left UF; and increased length of right AF and CC. Volume of bilateral UF and right AF and fiber density of left UF were positively associated with autistic features. PMID- 20019146 TI - The nature and time course of cortical activation following subthalamic stimulation in Parkinson's disease. AB - We studied the time course and nature of interactions between the subthalamic nucleus (STN) and the motor cortex in 8 Parkinson disease (PD) patients with chronically implanted STN deep-brain stimulation (DBS) electrodes. We first identified the cortical evoked potentials following STN stimulation. The most consistent potential was positive wave with peak latency of 22.2 +/- 1.2 ms from stimulation of clinically effective contacts. We then stimulated the motor cortex with transcranial magnetic stimulation (TMS) at 2-15 ms and at the latency of the evoked potential ( approximately 23 ms) following STN DBS. TMS induced currents in 3 directions: lateral-medial (LM) direction activated corticospinal axons directly, posterior-anterior (PA), and anterior-posterior (AP) directions activated corticospinal neurons transynaptically. Motor-evoked potentials (MEP) elicited by AP and PA TMS were facilitated at short (2-4 ms) and medium latencies (21-24 ms). However, MEPs elicited by LM TMS were not modified by STN DBS. Short latency antidromic stimulation of the corticosubthalamic projections and medium latency transmission likely through the basal ganglia-thalamocortical circuit led to cortical evoked potentials and increased motor cortex excitability at specific intervals following STN stimulation at clinically effective contacts. Cortical activation may be related to the clinical effects of STN DBS in PD. PMID- 20019148 TI - Relapse of enterococcal prosthetic valve endocarditis with aortic root abscess following treatment with daptomycin in a patient not fit for surgery. AB - Daptomycin is a novel lipopeptide with activity against Gram-positive organisms including enterococci. It is licensed for the treatment of Staphylococcus aureus bacteraemia and right-sided endocarditis, but not endocarditis due to Enterococcus spp. We report a case of enterococcal prosthetic valve endocarditis with an aortic root abscess in an elderly patient who was not fit for surgery. The patient's endocarditis relapsed 9 weeks after a 6 week course of daptomycin. PMID- 20019147 TI - Coding and binding of color and form in visual cortex. AB - The processing of color and form is largely segregated within the visual brain. But there is also evidence to suggest that these features are coded in combination early in visual processing. Here, we combined high-resolution functional magnetic resonance imaging (fMRI) together with multivariate pattern classification to examine where in the visual cortex specific color form "conjunctions" are represented. Human subjects viewed visual displays containing colored spiral patterns. The spiral patterns could be red or green, and oriented either clockwise or counterclockwise, leading to 4 possible stimulus configurations. Two additional displays combined 2 of the above single color-form pairings, leading to double conjunctions. We applied linear classifiers to voxel activation patterns obtained while subjects viewed such displays. Our findings not only show that color and form information is coded across retinotopically defined visual areas, but also that the 2 double-conjunction stimuli can be distinguished. The voxels most informative about conjunctions were distinct from those most informative about color or form alone. Our results indicate that conjunctions of form and color may be coded by separate functional units as early as primary visual cortex. The results of this study have implications for theories concerning the segregation and binding of color and form information. PMID- 20019149 TI - Fluctuations in phenotypes and genotypes within populations of Pseudomonas aeruginosa in the cystic fibrosis lung during pulmonary exacerbations. AB - Chronic respiratory infection by Pseudomonas aeruginosa contributes significantly to the morbidity and mortality associated with cystic fibrosis (CF). Using a series of phenotypic and genotypic tests on collections of 40 isolates per sputum sample, we analysed fluctuations within sputum populations of the P. aeruginosa Liverpool epidemic strain (LES) during pulmonary exacerbations. For each of three patients, three sequential sputum samples were analysed: (1) on presentation with exacerbation at the Regional Adult Cystic Fibrosis Unit, Liverpool; (2) a few days into intravenous antibiotic treatment; (3) when the patient had recovered. Fluctuations were observed in morphotype distribution, the production of virulence-associated quorum-sensing-dependent exoproducts (the phenazine compound pyocyanin and the elastase LasA), antibiotic susceptibility profiles and levels of auxotrophy. PCR assays were used to screen isolates for the presence of novel regions of the LES genome (islands and prophages) and to detect free phages. In one patient there was an increase in the prevalence of the LESGI-5 genomic island during the sampling period from 10 to 97.5 % carriage. LES phages 2-4 were detected in either the majority or all sputum samples tested, indicating widespread phage activity during the sampling period. The results of this study are indicative that significant fluctuations occur within P. aeruginosa populations during short periods of pulmonary exacerbation and intravenous antibiotic therapy. PMID- 20019150 TI - Reduced SERCA2 abundance decreases the propensity for Ca2+ wave development in ventricular myocytes. AB - AIMS: To describe the overall role of reduced sarcoplasmic reticulum Ca(2+) ATPase (SERCA2) for Ca(2+) wave development. METHODS AND RESULTS: SERCA2 knockout [Serca2(flox/flox) Tg(alphaMHC-MerCreMer); KO] mice allowing inducible cardiomyocyte-specific disruption of the Serca2 gene in adult mice were compared with Serca(flox/flox) (FF) control mice. Six days after Serca2 gene disruption, SERCA2 protein abundance was reduced by 53% in KO compared with FF, whereas SERCA2 activity in field-stimulated, Fluo-5F AM-loaded cells was reduced by 42%. Baseline Ca(2+) content of the sarcoplasmic reticulum (SR) and Ca(2+) transient amplitude and rate constant of decay measured in whole-cell voltage-clamped cells were decreased in KO to 75, 81, and 69% of FF values. Ca(2+) waves developed in only 31% of KO cardiomyocytes compared with 57% of FF when external Ca(2+) was raised (10 mM), although SR Ca(2+) content needed for waves to develop was 79% of FF values. In addition, waves propagated at a 15% lower velocity in KO cells. Ventricular extrasystoles (VES) occurred with lower frequency in SERCA2 KO mice (KO: 3 +/- 1 VES/h vs. FF: 8 +/- 1 VES/h) (P < 0.05 for all results). CONCLUSION: Reduced SERCA2 abundance resulted in decreased amplitude and decay rate of Ca(2+) transients, reduced SR Ca(2+) content, and decreased propensity for Ca(2+) wave development. PMID- 20019155 TI - Articles highlighted. PMID- 20019156 TI - Foxn1 gene knockout suppresses sexual attractiveness and pheromonal components of male urine in inbred mice. AB - The immunocompetence handicap hypothesis (ICHH) posits that females prefer signals emitted by immunocompetent males over immunocompromised males and that these signals are honest. However, mechanisms of mate choice under an ICHH model may be impacted by levels of genetic variation (inbred animals vs. outbred animals). Here, we conducted 2-choice female preference experiments and chemical analyses of male urine in inbred BALB/c and outbred CD-1 mice, both of which have immunocompromised nude (nu) strains resulting from a Foxn1 gene knockout. We found that inbred BALB/c females but not outbred CD-1 females preferred the urine of healthy males over that of immunocompromised males despite measured differences in the qualities of their urine. There was a clear increase in female attracting pheromones (such as farnesenes) in the preputial glands and urine metabolites in healthy BALB/c males but no such difference between CD-1 and CD-1 nu males. Therefore, CD-1 male urine failed to provide an honest mate-choice cue for females. Our results suggest that deleterious traits associated with male odor in mice might be jointly affected by the level of inbreeding and immunodeficiency caused by a single-gene knockout. PMID- 20019158 TI - Heterogeneity in human cardiovascular function contributes to a deeper understanding of integrative mechanisms. PMID- 20019159 TI - Tiny bubbles. PMID- 20019160 TI - Esophageal pressures in acute lung injury: do they represent artifact or useful information about transpulmonary pressure, chest wall mechanics, and lung stress? AB - Acute lung injury can be worsened by inappropriate mechanical ventilation, and numerous experimental studies suggest that ventilator-induced lung injury is increased by excessive lung inflation at end inspiration or inadequate lung inflation at end expiration. Lung inflation depends not only on airway pressures from the ventilator but, also, pleural pressure within the chest wall. Although esophageal pressure (Pes) measurements are often used to estimate pleural pressures in healthy subjects and patients, they are widely mistrusted and rarely used in critical illness. To assess the credibility of Pes as an estimate of pleural pressure in critically ill patients, we compared Pes measurements in 48 patients with acute lung injury with simultaneously measured gastric and bladder pressures (Pga and P(blad)). End-expiratory Pes, Pga, and P(blad) were high and varied widely among patients, averaging 18.6 +/- 4.7, 18.4 +/- 5.6, and 19.3 +/- 7.8 cmH(2)O, respectively (mean +/- SD). End-expiratory Pes was correlated with Pga (P = 0.0004) and P(blad) (P = 0.0104) and unrelated to chest wall compliance. Pes-Pga differences were consistent with expected gravitational pressure gradients and transdiaphragmatic pressures. Transpulmonary pressure (airway pressure - Pes) was -2.8 +/- 4.9 cmH(2)O at end exhalation and 8.3 +/- 6.2 cmH(2)O at end inflation, values consistent with effects of mediastinal weight, gravitational gradients in pleural pressure, and airway closure at end exhalation. Lung parenchymal stress measured directly as end-inspiratory transpulmonary pressure was much less than stress inferred from the plateau airway pressures and lung and chest wall compliances. We suggest that Pes can be used to estimate transpulmonary pressures that are consistent with known physiology and can provide meaningful information, otherwise unavailable, in critically ill patients. PMID- 20019161 TI - Intrinsic versus idiosyncratic drug-induced hepatotoxicity--two villains or one? AB - "Intrinsic" and "idiosyncratic" drug-induced liver injury reactions are commonly thought to arise by different modes of action. Intrinsic toxicity is reproducible in animals and occurs dose-dependently at sublethal doses. Environmental and genetic sensitivity factors can influence the toxicity of intrinsic hepatotoxicants. Among these is inflammatory stress. For example, exposure of mice to inflammatory bacterial lipopolysaccharide (LPS) causes a leftward shift in the dose-response relationship for acetaminophen hepatotoxicity; that is, acetaminophen toxicity is enhanced by LPS-induced inflammatory stress. Idiosyncratic reactions present themselves very differently than intrinsic ones; they happen in a minority of patients, with variable time of onset and no obvious relationship to drug dose, and they are not reproducible in usual animal tests. Although these characteristics seem to distinguish them from intrinsic reactions, consideration of fundamental principles of dose response can explain the differences. For a drug that causes idiosyncratic hepatotoxicity, the liver may not be a typical target for toxicity because the dose-response curve for hepatotoxicity lies to the right of the lethal dose. However, a sporadically occurring sensitivity factor, such as an inflammatory episode, could shift the dose-response curve for hepatotoxicity to the left, thereby bringing hepatotoxic doses into the therapeutic range. This hypothesis can account for the bizarre characteristics of idiosyncratic reactions and is supported by recent results showing that several drugs associated with human idiosyncratic reactions can be rendered hepatotoxic to rodents upon interaction with an inflammatory stimulus. In light of this view, intrinsic and idiosyncratic reactions may not be that different after all. PMID- 20019162 TI - Combined FISH and PRINS sperm analysis of complex chromosome rearrangement t(1;19;13): an approach facilitating PGD. AB - Complex chromosome rearrangements (CCRs) are structural aberrations involving three or more breakpoints on two or more chromosomes. These CCRs result in a high rate of chromosome imbalances potentially leading to subfertility and congenital abnormality. In this study, we analysed meiotic segregation in the sperm of a patient with a familial CCR 46, XY,t(1;19;13)(p31;q13.2;q31)mat included in an intracytoplasmic sperm injection program because of oligoasthenozoospermia. The rearrangement was first identified using conventional and molecular cytogenetic methods. Primed in situ labelling (PRINS) and fluorescence in situ hybridization (FISH) techniques were then combined allowing the simultaneous use of five fluorochromes on the same sperm preparation, for the segregation analysis and the evaluation of the reproductive options for this patient. Segregation analysis was performed in a total of 1822 sperm nuclei from the translocation carrier. The percentage of unbalanced sperm was 75.9%, including 34.1% from 3:3 segregation, 38.2% from 4:2 segregation, 3.5% from 5:1 segregation and 0.05% from 6:0 segregation. Only 14.8% of sperm nuclei were consistent with a normal or balanced chromosome complement. In conclusion, chromosome segregation analysis combining FISH and PRINS was performed in sperm from a CCR carrier using five fluorochromes. These results advance our understanding of the mechanisms of meiotic segregation, and facilitate the assessment of the usefulness of preimplantation genetic diagnosis procedures in CCR couples. PMID- 20019163 TI - Human hephaestin expression is not limited to enterocytes of the gastrointestinal tract but is also found in the antrum, the enteric nervous system, and pancreatic {beta}-cells. AB - Hephaestin (Hp) is a membrane protein with ferroxidase activity that converts Fe(II) to Fe(III) during the absorption of nutritional iron in the gut. Using anti-peptide antibodies to predicted immunogenic regions of rodent Hp, previous immunocytochemical studies in rat, mouse, and human gut tissues localized Hp to the basolateral membranes of the duodenal enterocytes where the Hp was predicted to aid in the transfer of Fe(III) to transferrin in the blood. We used a recombinant soluble form of human Hp to obtain a high-titer polyclonal antibody to Hp. This antibody was used to identify the intracellular location of Hp in human gut tissue. Our immunocytochemical studies confirmed the previous localization of Hp in human enterocytes. However, we also localized Hp to the entire length of the gastrointestinal tract, the antral portion of the stomach, and to the enteric nervous system (both the myenteric and submucous plexi). Hp was also localized to human pancreatic beta-cells. In addition to its expression in the same cells as Hp, ferroportin was also localized to the ductal cells of the exocrine pancreas. The localization of the ferroxidase Hp to the neuronal plexi and the pancreatic beta cells suggests a role for the enzymatic function of Hp in the protection of these specialized cell types from oxidative damage. PMID- 20019164 TI - Progesterone receptor A mediates VIP inhibition of contraction. AB - The slow transit time of the colon in females with constipation is due to impairment of agonist-induced contraction. The impairment is associated with downregulation of G proteins that mediate contraction and upregulation of Gs proteins that mediate relaxation. These changes are caused by overexpression of progesterone (P4) receptors in the colon, rendering its muscle cells sensitive to physiological P4 concentrations. Downregulation of Gq/11 is mediated by P4 receptor B (PR-B). We examined whether upregulation of Gs proteins increased the inhibition of contraction and whether the increase is mediated by the P4 receptor A (PR-A). These studies were conducted in colon-isolated colon muscle cells from human control and slow-transit constipation (STC) females and from guinea pigs. Muscle cell contraction was induced by CCK-8. Inhibition of contraction was induced by vasoactive intestinal polypeptide (VIP), and 8'bromo-c'AMP (8B-c'AMP) G protein levels were determined by Western blot. VIP-induced inhibition of contraction was greater in muscle cells from STC and P4-treated muscle cells. There were no differences in the inhibition induced by 8B-c'AMP between muscle cells from STC and P4-treated controls. The increased VIP-induced inhibition of muscle cells treated with P4 was blocked by pretreatment with PR-A antibodies and unaffected by PR-B antibodies. These antibodies had no effect on 8B-c'AMP induced inhibition. The P4 upregulation of Gs proteins was blocked by PR-A antibodies and unaffected by PR-B antibodies. Similar results were obtained in muscle cells from guinea pig colons. We concluded that P4 upregulation of Gs proteins increases VIP induced inhibition of contraction mediated by PR-A. PMID- 20019165 TI - Analysis of real-time serotonin (5-HT) availability during experimental colitis in mouse. AB - Serotonin (5-HT)-containing enterochromaffin (EC) cells of the intestine transduce chemical and mechanical stimuli from the intestinal lumen by releasing 5-HT on to afferent nerve terminals. Dysfunctional mucosal 5-HT signaling has been implicated in heightened visceral sensitivity and altered motility in patients with inflammatory bowel disease and in animal models. Our aim was to characterize the release and uptake of 5-HT in the mouse dextran sulfate sodium (DSS; 5% wt/vol) model of colitis. We made electrochemical recordings and used an ELISA assay to determine mucosal 5-HT release and uptake in untreated mice and mice with DSS-induced colitis. Peak and steady-state 5-HT concentrations were measured before and during blockade of the serotonin reuptake transporter (SERT) with 1 microM fluoxetine. Electrochemical recordings showed that colons from DSS treated mice had roughly twice the steady-state levels of extracellular 5-HT and compression-evoked 5-HT release compared with untreated mice. Fluoxetine doubled the compression-evoked and steady-state 5-HT levels in control and DSS mice. These data were supported by ELISA assays, which showed enhanced 5-HT release during colitis, by immunohistochemical analyses, which showed increases in EC cell numbers, and by real-time PCR, which identified a decrease in SERT mRNA expression in the mucosa during colitis. These data are the first to demonstrate 5-HT release close to its release site and near its site of action during DSS colitis. We conclude that DSS-colitis increases 5-HT availability primarily by an increase in the numbers of EC cells and/or of content of 5-HT in these EC cells. PMID- 20019166 TI - Dickkopf-1 promotes hyperglycemia-induced accumulation of mesangial matrix and renal dysfunction. AB - Wnt/beta-catenin signaling mediates renal fibrosis in several model systems including diabetic nephropathy. Dickkopf-1 (DKK-1) is an endogenous inhibitor of Wnt/beta-catenin signaling, but whether DKK-1 modulates diabetic nephropathy is unknown. Here, we studied whether DKK-1 participates in high glucose (HG)-induced expression of profibrotic factors and renal damage. In vitro, HG increased expression of DKK1, receptor Kremen-2, TGF-beta1, and fibronectin in mesangial cells. Loss and gain of DKK1 function modulated HG-mediated c-Jun, TGF-beta1, and fibronectin expression. DKK1 mediated HG-induced phosphorylation of Ser45-beta catenin and reduction of nuclear beta-catenin levels, but not phosphorylation of ERK kinase. Wnt3a protein and the beta-catenin (Delta45) mutation increased nuclear beta-catenin but abrogated HG-induced DKK1 and fibronectin expression. Exogenous DKK1 antisense oligonucleotide attenuated the increase in both serum DKK1 and urinary protein excretion in streptozotocin-induced diabetic rats. Knocking down DKK1 inhibited mesangial expression of TGF-beta1 and fibronectin and reduced both the glomerular volume and deposition of mesangial matrix in diabetic kidneys. Taken together, DKK1 mediates HG-induced destabilization of beta-catenin and matrix accumulation in mesangial cells. Knocking down DKK1 prevents diabetes-induced renal dysfunction and microstructure deterioration, suggesting that inhibition of DKK1offers therapeutic potential for diabetic nephropathy. PMID- 20019168 TI - Acute kidney injury associates with increased long-term mortality. AB - Acute kidney injury (AKI) associates with higher in-hospital mortality, but whether it also associates with increased long-term mortality is unknown, particularly after accounting for residual kidney function after hospital discharge. We retrospectively analyzed data from US veteran patients who survived at least 90 d after discharge from a hospitalization. We identified AKI events not requiring dialysis from laboratory data and classified them according to the ratio of the highest creatinine during the hospitalization to the lowest creatinine measured between 90 d before hospitalization and the date of discharge. We estimated mortality risks using multivariable Cox regression models adjusting for demographics, comorbidities, medication use, primary diagnosis of admission, length of stay, mechanical ventilation, and postdischarge estimated GFR (residual kidney function). Among the 864,933 hospitalized patients in the study cohort, we identified 82,711 hospitalizations of patients with AKI. In the study population of patients who survived at least 90 d after discharge, 17.4% died during follow-up (AKI 29.8%, without AKI 16.1%). The adjusted mortality risk associated with AKI was 1.41 (95% confidence interval [CI] 1.39 to 1.43) and increased with increasing AKI stage: 1.36 (95% CI 1.34 to 1.38), 1.46 (95% CI 1.42 to 1.50), and 1.59 (95% CI 1.54 to 1.65; P < 0.001 for trend). In conclusion, AKI that does not require dialysis associates with increased long term mortality risk, independent of residual kidney function, for patients who survive 90 d after discharge. Long-term mortality risk is highest among the most severe cases of AKI. PMID- 20019167 TI - New insights into epithelial-mesenchymal transition in kidney fibrosis. AB - Epithelial-mesenchymal transition (EMT), a process by which differentiated epithelial cells undergo a phenotypic conversion that gives rise to the matrix producing fibroblasts and myofibroblasts, is increasingly recognized as an integral part of tissue fibrogenesis after injury. However, the degree to which this process contributes to kidney fibrosis remains a matter of intense debate and is likely to be context-dependent. EMT is often preceded by and closely associated with chronic interstitial inflammation and could be an adaptive response of epithelial cells to a hostile or changing microenvironment. In addition to tubular epithelial cells, recent studies indicate that endothelial cells and glomerular podocytes may also undergo transition after injury. Phenotypic alteration of podocytes sets them in motion to functional impairment, resulting in proteinuria and glomerulosclerosis. Several intracellular signal transduction pathways such as TGFbeta/Smad, integrin-linked kinase (ILK) and Wnt/beta-catenin signaling are essential in controlling the process of EMT and presently are potential targets of antifibrotic therapy. This review highlights the current understanding of EMT and its underlying mechanisms to stimulate further discussion on its role, not only in the pathogenesis of renal interstitial fibrosis but also in the onset of podocyte dysfunction, proteinuria, and glomerulosclerosis. PMID- 20019169 TI - Tubular expression of KIM-1 does not predict delayed function after transplantation. AB - Injured epithelial cells of the proximal tubule upregulate the glycoprotein kidney injury molecule 1 (KIM-1), suggesting its potential as a biomarker of incipient kidney allograft injury. It is unknown whether KIM-1 expression changes in kidney allografts with delayed graft function (DGF), which often follows ischemia-reperfusion injury. Here, we prospectively measured KIM-1 RNA and protein expression in preperfusion biopsies of 30 living- and 85 deceased-donor kidneys and correlated the results with histologic and clinical outcomes after transplantation. We detected KIM-1 expression in 62% of deceased-donor kidneys and only 13% of living-donor kidneys (P < 0.0001). The level of KIM-1 expression before reperfusion correlated inversely with renal function at the time of procurement and correlated directly with the degree of interstitial fibrosis. Surprising, however, we did not detect a significant correlation between KIM-1 staining intensity and the occurrence of DGF. Our findings are consistent with a role for KIM-1 as an early indicator of tubular injury but do not support tissue KIM-1 measurement before transplantation to identify kidneys at risk for DGF. PMID- 20019170 TI - Incidence and epidemiology of levofloxacin resistance in Streptococcus pneumoniae: experience from a tertiary referral hospital in England. AB - OBJECTIVES: To investigate the incidence of levofloxacin resistance in Streptococcus pneumoniae isolates cultured by Lancashire Teaching Hospitals NHS Foundation Trust (LTHTR), and detect cases of in vivo resistance development. METHODS: During the study period (September 2004-February 2007), isolates of S. pneumoniae cultured by the LTHTR microbiology laboratory were examined by Etest to determine MICs of levofloxacin. Isolates from patients in whom there was a shift towards colonization with S. pneumoniae of reduced levofloxacin susceptibility were further characterized by serotyping, multilocus sequence typing (MLST) and sequencing of parC and gyrA genes. RESULTS: Eight hundred and sixty-five isolates were collected; however, 772 isolates from 652 patients were recoverable; 412 (53.4%) came from hospitalized patients, 12 (1.6%) were resistant to levofloxacin according to the BSAC breakpoint (>2 mg/L) and 29 (3.8%) had MICs at the breakpoint (MIC = 2 mg/L). Of six patients in whom there was a shift towards isolates with reduced levofloxacin susceptibility, five had acquired new distinct strains. One patient, who had a parC mutation (Ser79Phe) in the original susceptible isolate and an additional second-step mutation in the gyrA gene (Ser81Phe) of the later resistant one, had isolates belonging to the same pneumococcal clone. CONCLUSIONS: S. pneumoniae resistance to levofloxacin was uncommon and we managed to identify only one case of probable in vivo resistance development in the 2.5 years of the study. Strain replacement accounted for the majority of incidences where there was an apparent shift towards colonization with isolates of reduced levofloxacin susceptibility. PMID- 20019171 TI - A 12 month clinical audit of cervical spine imaging in multiply injured and intubated patients. AB - Previous work has questioned how plain films should be used when imaging the cervical spine of trauma patients. The authors wanted to identify whether the National Institute for Clinical Excellence (NICE) guidelines were being followed with respect to the imaging of patients presenting with cervical spine injury over a 1 year period. Data retrieved from the Electronic Digital Information Service (EDIS) computerised database records of all patients presenting with a triage code 1 or 2 between 1 September 2007 and 31 August 2008 were used to conduct a retrospective audit that identified multiply injured and intubated patients who did not undergo CT of the cervical spine and to highlight the use of plain films when the patient was to undergo CT of the head and cervical spine. A clinical record search identified 52 patients with a mean age of 32 years, of whom 73% were males, who had been admitted with multiple traumas and had undergone imaging of the cervical spine. Although no patient was intubated without undergoing CT of the cervical spine or head, seven patients had plain films when it was clear that they were to undergo CT. In conclusion, the audit emphasised the excellent work of emergency department and radiology staff in identifying and imaging multiple trauma patients, as all patients requiring CT of the cervical spine received this investigation. However, careful thought should be given to ordering plain films before CT, as some patients who clearly required CT of the cervical spine underwent unnecessary lateral plain films in the emergency department, delaying their progression to definitive care. PMID- 20019172 TI - A combination of cisplatin-eluting gelatin microspheres and flavopiridol enhances anti-tumour effects in a rabbit VX2 liver tumour model. AB - The aim of this study was to investigate whether the combination of cisplatin eluting gelatin microspheres (GMSs) and flavopiridol enhances anti-tumour effects in a rabbit VX2 liver tumour model. Tumour-bearing rabbits (n = 21) were divided into five groups and infused from the proper hepatic artery. Group 1 (n = 5) received cisplatin-eluting GMSs (1 mg kg(-1)) and flavopiridol (3 mg kg(-1)), group 2 (n = 5) cisplatin-eluting GMSs alone (1 mg kg(-1)), Group 3 (n = 5) flavopiridol (3 mg kg(-1)), Group 4 (n = 3) GMSs alone (1 mg kg(-1)), and Group 5 (n = 3) was the control group receiving physiological saline (1 ml kg(-1)). On days 0 and 7 after procedures the liver tumour volume was measured using a horizontal open MRI system and the relative tumour volume growth rates for 7 days after treatment were calculated. On T(1) weighted images, the tumours were visualised as circular, low-intensity areas just below the liver surface. After treatment, the signals remained similar. The relative tumour volume growth rate for 7 days after treatment was 54.2+/-22.4% in Group 1, 134.1+/-40.1% in Group 2,166.7+/-48.1% in Group 3, 341.8+/-8.6% in Group 4 and 583.1+/-46.9% in Group 5; the growth rate was significantly lower in Group 1 than the other groups (p<0.05). We concluded that in our rabbit model of liver tumours the combination of cisplatin-eluting GMSs and flavopiridol was effective. PMID- 20019173 TI - Investigation of the effect of anode/filter materials on the dose and image quality of a digital mammography system based on an amorphous selenium flat panel detector. AB - A comparison, in terms of image quality and glandular breast dose, was carried out between two similar digital mammography systems using amorphous selenium flat panel detectors. The two digital mammography systems currently available from Lorad-Hologic were compared. The original system utilises Mo/Mo and Mo/Rh as target/filter combinations, while the new system uses W/Rh and W/Ag. Images of multiple mammography phantoms with simulated compressed breast thicknesses of 4 cm, 5 cm and 6 cm and various glandular tissue equivalency were acquired under different spectral conditions. The contrast of five details, corresponding to five glandular compositions, was calculated and the ratio of the square of the contrast-to-noise ratio to the average glandular dose was used as a figure-of merit (FOM) to compare results. For each phantom thickness and target/filter combination, there is an optimum voltage that maximises the FOM. Results show that the W/Rh combination is the best choice for all the detection tasks studied, but for thicknesses greater than 6 cm the W/Ag combination would probably be the best choice. In addition, the new system with W filter presents a better optimisation of the automatic exposure control in comparison with the original system with Mo filter. PMID- 20019174 TI - Simplified rules for everyday delineation of lymph node areas for breast cancer radiotherapy. AB - The aim of this study was to present the simplified rules of delineation of lymph node (LN) volumes in breast irradiation. Practical rules of delineation of LN areas were developed in the Department of Radiation Oncology of the Institut Curie. These practical guidelines of delineation were based on different specific publications in the field of breast and LN anatomy. The principal characteristic of these rules is their clearly established relationship with anatomical structure, which is easy to find on CT slices. The simplified rules of delineation have been published in pocket format as the illustrated atlas "Help of delineation for breast cancer treatment". In this small pocket guide, delineation using the practical rules is illustrated, with examples from anatomical CT slices. It is shown that there is an improvement in delineation after the use of these simplified rules and the guide. In conclusion, this small guide is useful for improving everyday practice and decreasing the differences in target delineation for breast irradiation between institutions and observers. PMID- 20019175 TI - Impact of biplane versus single-plane imaging on radiation dose, contrast load and procedural time in coronary angioplasty. AB - Coronary angioplasties can be performed with either single-plane or biplane imaging techniques. The aim of this study was to determine whether biplane imaging, in comparison to single-plane imaging, reduces radiation dose and contrast load and shortens procedural time during (i) primary and elective coronary angioplasty procedures, (ii) angioplasty to the main vascular territories and (iii) procedures performed by operators with various levels of experience. This prospective observational study included a total of 504 primary and elective single-vessel coronary angioplasty procedures utilising either biplane or single-plane imaging. Radiographic and clinical parameters were collected from clinical reports and examination protocols. Radiation dose was measured by a dose-area-product (DAP) meter intrinsic to the angiography system. Our results showed that biplane imaging delivered a significantly greater radiation dose (181.4+/-121.0 Gycm(2)) than single-plane imaging (133.6+/-92.8 Gycm(2), p<0.0001). The difference was independent of case type (primary or elective) (p = 0.862), vascular territory (p = 0.519) and operator experience (p = 0.903). No significant difference was found in contrast load between biplane (166.8+/-62.9 ml) and single-plane imaging (176.8+/-66.0 ml) (p = 0.302). This non-significant difference was independent of case type (p = 0.551), vascular territory (p = 0.308) and operator experience (p = 0.304). Procedures performed with biplane imaging were significantly longer (55.3+/-27.8 min) than those with single-plane (48.9+/-24.2 min, p = 0.010) and, similarly, were not dependent on case type (p = 0.226), vascular territory (p = 0.642) or operator experience (p = 0.094). Biplane imaging resulted in a greater radiation dose and a longer procedural time and delivered a non-significant reduction in contrast load than single-plane imaging. These findings did not support the commonly perceived advantages of using biplane imaging in single-vessel coronary interventional procedures. PMID- 20019176 TI - The developing role for intensity-modulated radiation therapy (IMRT) in the non surgical treatment of brain metastases. AB - Whole brain radiotherapy (WBRT) is the standard non-surgical treatment for brain metastatic disease, but rarely eradicates bulky metastases from most common cancers. Recent literature has demonstrated the safety and efficacy of delivering very high focal doses of radiation (by radiosurgical techniques) to the gross tumour volume of bulky brain metastases, thereby obtaining more certain local control than is achieved by WBRT. In this paper we report a study of 11 patients with bulky brain metastases in whom an intensity-modulated radiation therapy (IMRT) facility has been used to concomitantly boost the gross tumour volume of bulky brain metastatic disease (to 40 Gy) during a standard 30 Gy in 10 fractions WBRT schedule. No acute or subacute morbidity was encountered, and good early control data were noted. We discuss the perceived advantages of such a technique. PMID- 20019177 TI - The apparent increase in the {beta}-parameter of the linear quadratic model with increased linear energy transfer during fast neutron irradiation. AB - The issue of whether the beta-parameter of the linear quadratic model changes with linear energy transfer (LET) remains controversial. Retrospective analysis of UK fast neutron experimental data using human cell lines at Clatterbridge shows that the beta-parameter of the linear quadratic model probably does increase with LET during neutron irradiation. For cells without a deficiency in DNA damage repair and for experiments in which beta-parameter estimates were considered to be unreliably low, a provisional relationship of beta(H) = 1.82 beta(L) was found (where the suffixes refer to high and low LET exposures, respectively). This implies that radicalbeta increases by around 1.35 in the specific case of 62.5 MeV neutrons relative to 4 MeV X-rays. Increments in the beta-parameter with LET influence the relative biological effect (RBE), especially at high doses per fraction. Large fractions are being used in experimental carbon ion therapy, in which broadly similar RBE values to fast neutrons are found. These interesting findings after fast neutron exposure need to be studied further for applications in charged particle beam therapy using light ions, which is presently undergoing a worldwide expansion. PMID- 20019179 TI - The effects of positive and negative support from children on widowed older adults' psychological adjustment: a longitudinal analysis. AB - PURPOSE: This article examines the extent to which positive and negative support from children prior to and after spousal loss and changes in support from pre- to post-loss affect widowed older adults' depressive symptoms, anxiety, and anger 18 months following widowhood. DESIGN AND METHODS: Analyses are based on the Changing Lives of Older Couples, a prospective study of 1,532 married individuals aged 65 years and older. The analytic sample includes 148 widowed persons who have at least one living child and participated in the baseline and two follow-up interviews conducted 6 and 18 months following widowhood. RESULTS: The analyses revealed that higher levels of positive support from children 6 months following widowhood were associated with fewer depressive symptoms at the 18-month follow up, whereas higher levels of negative support 18 months after loss and a decrease in positive support following loss were associated with more depressive symptoms. Greater negative support both prior to and after loss was associated with greater anxiety, whereas greater negative support 6 months after loss and an increase in negative support following loss were related to greater anger. IMPLICATIONS: Given their differential impact on widowed persons' psychological adjustment, both positive and negative aspects of social support as well as its changing nature should be considered in working with bereaved older adults. PMID- 20019180 TI - Physical function decline and the risk of elder self-neglect in a community dwelling population. AB - PURPOSE: This longitudinal study examines the association between physical function decline and the risk of elder self-neglect in a community-dwelling population. DESIGN AND METHODS: Of the 5,570 participants in the Chicago Health Aging Project, 1,068 were reported to social services agency for suspected elder self-neglect from 1993 to 2005. The primary predictor was objectively assessed physical function using decline in physical performance testing. Secondary predictors were assessed using the decline in self-reported Katz, Nagi, and Rosow Breslau scales. Outcome of interest was elder self-neglect. Logistic and linear regression models were used to assess these associations. RESULTS: After adjusting for confounding factors, every 1-point decline in physical performance testing was associated with increased risk of reported elder self-neglect (odds ratio [OR], 1.05, confidence interval [CI], 1.03-1.07, p < .001). Decline in Katz (OR, 1.05, CI, 1.00-1.10, p < .05) and decline in Rosow-Breslau (OR, 1.19, CI, 1.11-1.27, p < .001) were associated with increased risk of reported elder self neglect. Decline in physical performance testing (standardized parameter estimate [PE]: 0.19, SE: 0.06, p = .002), Katz (PE: 0.65, SE: 0.14, p < .001), Nagi (PE: 0.48, SE: 0.14, p < .001), and Rosow-Breslau (PE: 0.57, SE: 0.21, p = .006) scales were associated with increased risk of greater self-neglect severity. IMPLICATIONS: Decline in physical function was associated with increased risk of reported elder self-neglect and greater self-neglect severity in this community dwelling population. PMID- 20019181 TI - The importance of ERbeta signalling in the ovary. AB - This review examines the evidence for a central role of oestrogen receptor beta (ERbeta or ESR2 as listed in the MGI Database) in folliculogenesis and hence reproductive biology. Knockout mouse models have been a valuable resource in this respect. The ERbeta-null mouse exhibits a granulosa cell phenotype associated with the partial arrest of folliculogenesis and ovulatory dysfunction. Phyto oestrogens such as genistein, which preferentially activate ERbeta, have been shown to alleviate the ovarian phenotype of the oestrogen-depleted aromatase knockout mouse. In normal adult mice, genistein has been shown to cause reproductive defectives following neonatal administration. Studies of ovarian cancer have also informed the literature. A decline in ERbeta levels in epithelial ovarian cancers has been hypothesised to be associated with severity of disease and prognosis. Whereas the abundant expression of ERbeta in granulosa cell tumours (GCT) of the ovary and evidence that ERbeta signalling is transrepressed by the nuclear factor-kappaB pathway in GCT cell lines suggest a pathogenetic role for ERbeta in GCT. In recent years, studies into the impact of environmental oestrogens (either in the form of pesticides or plastics) on reproductive function have shown that ERbeta-selective toxins cause reproductive dysfunction and impair fertility. It remains to be established as to what genes are regulated by ERbeta in the ovary. Finally, ERbeta has been shown to be regulated by gonadotrophins, the pituitary hormones mediating ovarian function. PMID- 20019182 TI - Mice lacking dystrophin or alpha sarcoglycan spontaneously develop embryonal rhabdomyosarcoma with cancer-associated p53 mutations and alternatively spliced or mutant Mdm2 transcripts. AB - Altered expression of proteins in the dystrophin-associated glycoprotein complex results in muscular dystrophy and has more recently been implicated in a number of forms of cancer. Here we show that loss of either of two members of this complex, dystrophin in mdx mice or alpha sarcoglycan in Sgca(-/-) mice, results in the spontaneous development of muscle-derived embryonal rhabdomyosarcoma (RMS) after 1 year of age. Many mdx and Sgca(-/-) tumors showed increased expression of insulin-like growth factor 2, retinoblastoma protein, and phosphorylated Akt and decreased expression of phosphatase and tensin homolog gene, much as is found in a human RMS. Further, all mdx and Sgca(-/-) RMS analyzed had increased expression of p53 and murine double minute (mdm)2 protein and contained missense p53 mutations previously identified in human cancers. The mdx RMS also contained missense mutations in Mdm2 or alternatively spliced Mdm2 transcripts that lacked an exon encoding a portion of the p53-binding domain. No Pax3:Fkhr or Pax7:Fkhr translocation mRNA products were evident in any tumor. Expression of natively glycosylated alpha dystroglycan and alpha sarcoglycan was reduced in mdx RMS, whereas dystrophin expression was absent in almost all human RMS, both for embryonal and alveolar RMS subtypes. These studies show that absence of members of the dystrophin-associated glycoprotein complex constitutes a permissive environment for spontaneous development of embryonal RMS associated with mutation of p53 and mutation or altered splicing of Mdm2. PMID- 20019183 TI - Mammalian target of rapamycin (mTOR) induces proliferation and de-differentiation responses to three coordinate pathophysiologic stimuli (mechanical strain, hypoxia, and extracellular matrix remodeling) in rat bladder smooth muscle. AB - Maladaptive bladder muscle overgrowth and de-differentiation in human bladder obstructive conditions is instigated by coordinate responses to three stimuli: mechanical strain, tissue hypoxia, and extracellular matrix remodeling.( 1,2) Pathway analysis of genes induced by obstructive models of injury in bladder smooth muscle cells (BSMCs) identified a mammalian target of rapamycin (mTOR) specific inhibitor as a potential pharmacological inhibitor. Strain-induced mTOR specific S6K activation segregated differently from ERK1/2 activation in intact bladder ex vivo. Though rapamycin's antiproliferative effects in vascular smooth muscle cells are well known, its effects on BSMCs were previously unknown. Rapamycin significantly inhibited proliferation of BSMCs in response to mechanical strain, hypoxia, and denatured collagen. Rapamycin inhibited S6K at mTOR-sensitive phosphorylation sites in response to strain and hypoxia. Rapamycin also supported smooth muscle actin expression in response to strain or hypoxia induced de-differentiation. Importantly, strain plus hypoxia synergistically augmented mTOR-dependent S6K activation, Mmp7 expression and proliferation. Forced expression of wild-type and constitutively active S6K resulted in loss of smooth muscle actin expression. Decreased smooth muscle actin, increased Mmp7 levels and mTOR pathway activation during in vivo partial bladder obstruction paralleled our in vitro studies. These results point to a coordinate role for mTOR in BSMCs responses to the three stimuli and a potential new therapeutic target for myopathic bladder disease. PMID- 20019185 TI - Memantine: "hypothesis testing" not "disease modifying" in Alzheimer's disease. PMID- 20019184 TI - Liver regeneration after partial hepatectomy: critical analysis of mechanistic dilemmas. AB - Liver regeneration after partial hepatectomy is one of the most studied models of cell, organ, and tissue regeneration. The complexity of the signaling pathways initiating and terminating this process have provided paradigms for regenerative medicine. Many aspects of the signaling mechanisms involved in hepatic regeneration are under active investigation. The purpose of this review is to focus on the areas still not well understood. The review also aims to provide insights into the ways by which current concepts of liver regeneration can provide understanding regarding malfunction of the regenerative process in liver diseases, such as acute liver failure. PMID- 20019186 TI - Liver-specific beta-catenin knockout mice exhibit defective bile acid and cholesterol homeostasis and increased susceptibility to diet-induced steatohepatitis. AB - Although the role of Wnt/beta-catenin signaling in liver growth and development is well established, its contribution in non-neoplastic hepatic pathologies has not been investigated. Here, we examine the role of beta-catenin in a murine model of diet-induced liver injury. Mice with hepatocyte-specific beta-catenin deletion (KO) and littermate controls were fed the steatogenic methionine and choline-deficient (MCD) diet or the corresponding control diet for 2 weeks and characterized for histological, biochemical, and molecular changes. KO mice developed significantly higher steatohepatitis and fibrosis on the MCD diet compared with wild-type mice. Both wild-type and KO livers accumulated triglyceride on the MCD diet but, unexpectedly, higher hepatic cholesterol levels were observed in KO livers on both control and MCD diets. Gene expression analysis showed that hepatic cholesterol accumulation in KO livers was not attributable to increased synthesis or uptake. KO mice had lower expression of bile acid synthetic enzymes but exhibited higher hepatic bile acid and serum bilirubin levels, suggesting defects in bile export. Therefore, loss of beta catenin in the liver leads to defective cholesterol and bile acid metabolism in the liver and increased susceptibility to developing steatohepatitis in the face of metabolic stress. PMID- 20019187 TI - Reduced expression of integrin alphavbeta8 is associated with brain arteriovenous malformation pathogenesis. AB - Brain arteriovenous malformations (BAVMs) are a rare but potentially devastating hemorrhagic disease. Transforming growth factor-beta signaling is required for proper vessel development, and defective transforming growth factor-beta superfamily signaling has been implicated in BAVM pathogenesis. We hypothesized that expression of the transforming growth factor-beta activating integrin, alphavbeta8, is reduced in BAVMs and that decreased beta8 expression leads to defective neoangiogenesis. We determined that beta8 protein expression in perivascular astrocytes was reduced in human BAVM lesional tissue compared with controls and that the angiogenic response to focal vascular endothelial growth factor stimulation in adult mouse brains with local Cre-mediated deletion of itgb8 and smad4 led to vascular dysplasia in newly formed blood vessels. In addition, common genetic variants in ITGB8 were associated with BAVM susceptibility, and ITGB8 genotypes associated with increased risk of BAVMs correlated with decreased beta8 immunostaining in BAVM tissue. These three lines of evidence from human studies and a mouse model suggest that reduced expression of integrin beta8 may be involved in the pathogenesis of sporadic BAVMs. PMID- 20019188 TI - S100A4 and metastasis: a small actor playing many roles. AB - The calcium-binding protein S100A4 promotes metastasis in several experimental animal models, and S100A4 protein expression is associated with patient outcome in a number of tumor types. S100A4 is localized in the nucleus, cytoplasm, and extracellular space and possesses a wide range of biological functions, such as regulation of angiogenesis, cell survival, motility, and invasion. In this review, we summarize the evidence connecting S100A4 and cancer metastasis and discuss the mechanisms by which S100A4 promotes tumor progression. PMID- 20019189 TI - Iron-dependent regulation of MDM2 influences p53 activity and hepatic carcinogenesis. AB - Iron overload is a risk factor for hepatocarcinoma, but the pathways involved are poorly characterized. Gene expression analysis in immortalized mouse hepatocytes exposed to iron or the iron chelator deferoxamine revealed that iron downregulated, whereas deferoxamine upregulated, mRNA levels of mouse double minute gene 2 (MDM2), the ubiquitin ligase involved in the degradation of the oncosuppressor p53. Regulation of MDM2 by iron status was observed at protein levels in mouse hepatocytes and rat liver, and was associated with specular changes in p53 expression. Iron dependent regulation of MDM2/p53 was confirmed ex vivo in human monocytes, by manipulation of iron pool and in a genetic model of iron deficiency, leading to modulation of p53 target genes involved in the antioxidant response and apoptosis. Iron status influenced p53 ubiquitination and degradation rate, and the MDM2 inhibitor nutlin increased p53 levels in iron depleted cells. Furthermore, nutlin enhanced the antiproliferative activity of deferoxamine in HepG2 hepatoblastoma cells. The MDM2 -309T > G promoter polymorphism, determining increased MDM2 and lower p53 activity, was associated with higher risk of hepatocarcinoma in cirrhotic patients with hemochromatosis, and with HFE mutations in patients with hepatocarcinoma without hemochromatosis, suggesting an interaction between MDM2 and iron in the pathogenesis of hepatocarcinoma. In conclusion, iron status influences p53 activity and antioxidant response by modulating MDM2 expression. MDM2 inhibitors may enhance the antiproliferative activity of iron chelators. PMID- 20019190 TI - The Sex-determining region Y-box 4 and homeobox C6 transcriptional networks in prostate cancer progression: crosstalk with the Wnt, Notch, and PI3K pathways. AB - The transforming growth factor beta, Hedgehog, Notch, and Wnt signaling pathways all play critical roles in the development and progression of prostate cancer. It is becoming increasingly apparent that these pathways may intersect with developmentally important transcription factors such as the sex-determining region Y-box 4 (SOX4), homeobox C6, enhancer of zeste 2, and ETS-related gene, which are up-regulated in prostate cancers. For example, identification of the downstream targets of SOX4 and homeobox C6 suggests that these factors may cooperate to activate the Notch pathway and the PI3K/AKT pathway, possibly in response to Wnt signals. PI3K/AKT activation likely occurs indirectly via up regulation of growth factor receptors, while Notch activation is secondary to up regulation of Notch pathway components. In addition, SOX4 may affect terminal differentiation via regulation of other transcription factors such as NKX3.1 and MLL, and regulation of components of the microRNA pathway such as Dicer and Argonaute 1. The evidence supporting activation of these pathways in prostate cancer progression suggests that combinations of compounds targeting them may be of benefit to patients with aggressive, metastatic disease. PMID- 20019191 TI - Human nephrosclerosis triggers a hypoxia-related glomerulopathy. AB - In the kidney, hypoxia contributes to tubulointerstitial fibrosis, but little is known about its implications for glomerular damage and glomerulosclerosis. Chronic hypoxia was hypothesized to be involved in nephrosclerosis (NSC) or "hypertensive nephropathy." In the present study genome-wide expression data from microdissected glomeruli were studied to examine the role of hypoxia in glomerulosclerosis of human NSC. Functional annotation analysis revealed prominent regulation of hypoxia-associated biological processes in NSC, including angiogenesis, fibrosis, and inflammation. Glomerular expression levels of a majority of genes regulated by the hypoxia-inducible factors (HIFs) were significantly altered in NSC. Among these HIF targets, chemokine C-X-C motif receptor 4 (CXCR4) was prominently induced. Glomerular CXCR4 mRNA induction was confirmed by quantitative RT-PCR in an independent cohort with NSC but not in those with other glomerulopathies. By immunohistological analysis, CXCR4 showed enhanced positivity in podocytes in NSC biopsy specimens. This CXCR4 positivity was associated with nuclear localization of HIF1alpha only in podocytes of NSC, indicating transcriptional activity of HIF. As the CXCR4 ligand CXCL12/SDF-1 is constitutively expressed in podocytes, autocrine signaling may contribute to NSC. In addition, a blocking CXCR4 antibody caused significant inhibition of wound closure by podocytes in an in vitro scratch assay. These data support a role for CXCR4/CXCL12 in human NSC and indicate that hypoxia not only is involved in tubulointerstitial fibrosis but also contributes to glomerular damage in NSC. PMID- 20019192 TI - Plasmepsin 4-deficient Plasmodium berghei are virulence attenuated and induce protective immunity against experimental malaria. AB - Plasmodium parasites lacking plasmepsin 4 (PM4), an aspartic protease that functions in the lysosomal compartment and contributes to hemoglobin digestion, have only a modest decrease in the asexual blood-stage growth rate; however, PM4 deficiency in the rodent malaria parasite Plasmodium berghei results in significantly less virulence than that for the parental parasite. P. berghei Deltapm4 parasites failed to induce experimental cerebral malaria (ECM) in ECM susceptible mice, and ECM-resistant mice were able to clear infections. Furthermore, after a single infection, all convalescent mice were protected against subsequent parasite challenge for at least 1 year. Real-time in vivo parasite imaging and splenectomy experiments demonstrated that protective immunity acted through antibody-mediated parasite clearance in the spleen. This work demonstrates, for the first time, that a single Plasmodium gene disruption can generate virulence-attenuated parasites that do not induce cerebral complications and, moreover, are able to stimulate strong protective immunity against subsequent challenge with wild-type parasites. Parasite blood-stage attenuation should help identify protective immune responses against malaria, unravel parasite-derived factors involved in malarial pathologies, such as cerebral malaria, and potentially pave the way for blood-stage whole organism vaccines. PMID- 20019193 TI - Multi-step aberrant CpG island hyper-methylation is associated with the progression of adult T-cell leukemia/lymphoma. AB - Aberrant CpG island methylation contributes to the pathogenesis of various malignancies. However, little is known about the association of epigenetic abnormalities with multistep tumorigenic events in adult T cell leukemia/lymphoma (ATLL). To determine whether epigenetic abnormalities induce the progression of ATLL, we analyzed the methylation profiles of the SHP1, p15, p16, p73, HCAD, DAPK, hMLH-1, and MGMT genes by methylation specific PCR assay in 65 cases with ATLL patients. The number of CpG island methylated genes increased with disease progression and aberrant hypermethylation in specific genes was detected even in HTLV-1 carriers and correlated with progression to ATLL. The CpG island methylator phenotype (CIMP) was observed most frequently in lymphoma type ATLL and was also closely associated with the progression and crisis of ATLL. The high number of methylated genes and increase of CIMP incidence were shown to be unfavorable prognostic factors and correlated with a shorter overall survival by Kaplan-Meyer analysis. The present findings strongly suggest that the multistep accumulation of aberrant CpG methylation in specific target genes and the presence of CIMP are deeply involved in the crisis, progression, and prognosis of ATLL, as well as indicate the value of CpG methylation and CIMP for new diagnostic and prognostic biomarkers. PMID- 20019194 TI - New look at an old problem: bacterial superinfection after influenza. PMID- 20019195 TI - Aberrant inflammatory response to Streptococcus pyogenes in mice lacking myeloid differentiation factor 88. AB - Several in vitro studies have emphasized the importance of toll-like receptor/myeloid differentiation factor 88 (MyD88) signaling in the inflammatory response to Streptococcus pyogenes. Since the extent of inflammation has been implicated in the severity of streptococcal diseases, we have examined here the role of toll-like receptor/MyD88 signaling in the pathophysiology of experimental S. pyogenes infection. To this end, we compared the response of MyD88-knockout (MyD88(-/-)) after subcutaneous inoculation with S. pyogenes with that of C57BL/6 mice. Our results show that MyD88(-/-) mice harbored significantly more bacteria in the organs and succumbed to infection much earlier than C57BL/6 animals. Absence of MyD88 resulted in diminished production of inflammatory cytokines such as interleukin-12, interferon-gamma, and tumor necrosis factor-alpha as well as chemoattractants such as monocyte chemotactic protein-1 (MCP-1) and Keratinocyte derived chemokine (KC), and hampered recruitment of effector cells involved in bacterial clearance (macrophages and neutrophils) to the infection site. Furthermore, MyD88(-/-) but not C57BL/6 mice exhibited a massive infiltration of eosinophils in infected organs, which can be explained by an impaired production of the regulatory chemokines, gamma interferon-induced monokine (MIG/CXCL9) and interferon-induced protein 10 (IP-10/CXCL10), which can inhibit transmigration of eosinophils. Our results indicate that MyD88 signaling targets effector cells to the site of streptococcal infection and prevents extravasation of cells that can induce tissue damage. Therefore, MyD88 signaling may be important for shaping the quality of the inflammatory response elicited during infection to ensure optimal effector functions. PMID- 20019197 TI - Promoting physical activity for persons with diabetes. AB - PURPOSE: The purpose of this study was to evaluate the effectiveness of a self help physical activity program for persons with type 2 diabetes. Effectiveness was measured by cardiovascular indicators, hemoglobin A1C, anthropometric indicators, and activity levels. METHODS: This intervention study included 53 individuals with type 2 diabetes. Participants were randomly assigned. Participants in the intervention group attended the usual Diabetes Self Management Education Program (DSMEP) and received a copy of the book, Manpo-kei; a concise handout summarizing the key points of Manpo-kei; and a pedometer. Participants not assigned to the intervention attended the usual DSMEP only. Data collection was completed at the beginning of the study and 3 months later using a lifestyle and diabetes questionnaire, a physical activity questionnaire, and cardiovascular and anthropometric measures. RESULTS: Those who participated in the intervention demonstrated a significant decrease in hemoglobin A1C, weight, and body fat. In addition, they increased their number of daily steps as measured by the pedometer. The control group also demonstrated a significant decrease in hemoglobin A1C and weight. Both groups demonstrated increases in their regular weekly activity. At the end of the study period, the intervention group had a significantly lower diastolic blood pressure than the control group. CONCLUSIONS: Pedometers can be a helpful strategy to motivate persons with diabetes to increase physical activity. However, it appears that attendance at DSMEP alone may be enough to increase physical activity for persons with type 2 diabetes. PMID- 20019196 TI - Chlamydophila pneumoniae infection leads to smooth muscle cell proliferation and thickening in the coronary artery without contributions from a host immune response. AB - Chlamydophila pneumonia (C. pneumonia) infection has been associated with the progression of atherosclerosis. It remains unclear, however, whether C. pneumoniae in the absence of an immune response can alone initiate atherogenic events within a complex vessel environment. Left anterior descending coronary arteries isolated from porcine hearts were dissected and placed in culture medium for 72 hours before infection with C. pneumoniae. C. pneumoniae replicated within the arterial wall for the duration of the experiment (up to 10 days). A significant increase in chlamydial-HSP60 protein expression from day 2 to 10 post infection (pi) indicated the presence of metabolically active C. pneumonia within infected vessels. Significant arterial thickening in infected coronary segments was observed by a considerable decrease in the ratio of lumen to total vessel area (48 +/- 3% at day 4 pi versus 23 +/- 3% at day 10 pi) and a significant increase in the ratio of media to luminal area (113 +/- 16% at day 4 pi versus 365 +/- 65% at day 10 pi). Structural changes were accompanied by an up regulation of host HSP60 and proliferating cell nuclear antigen expression levels. Immunohistochemical staining confirmed proliferating cell nuclear antigen expression to be primarily localized within smooth muscle cells of the medial area. These results demonstrate that C. pneumoniae infection can stimulate arterial thickening in a complex vessel environment without the presence of a host immune response and further supports the involvement of HSP60 in this action. PMID- 20019198 TI - Fetal cardiac screening: why bother? AB - Congenital heart disease (CHD) is a common form of congenital malformation associated with significant morbidity and mortality. Antenatal diagnosis of life threatening forms of CHD may help to improve survival and morbidity as well as allowing parental choice and preparation. The diagnosis of CHD during fetal life can be made with a very high level of diagnostic accuracy in tertiary centres dealing with high-risk pregnancies. However, most cases of CHD will occur in low risk groups and will only be detected by screening at the time of obstetric ultrasound scans. The concept of antenatal screening for CHD was introduced in the UK over 20 years ago, and current national guidelines recommend that the heart should be examined at the time of the obstetric anomaly scan. However, there remains a large regional variation in prenatal detection rates of CHD. Widespread teaching and training is required to achieve a more uniform standard but this will be associated with significant resource implications. PMID- 20019199 TI - Perspective on meconium staining of the amniotic fluid. PMID- 20019200 TI - Duration of meconium passage in preterm and term infants. PMID- 20019201 TI - Cord blood, perinatal BNP values in term and preterm newborns. PMID- 20019202 TI - Ischaemic intestinal disease in neonates with congenital heart defects: the timing of cardiac surgery does matter. PMID- 20019203 TI - PDA and pulmonary hypertension: should the duct be ligated? PMID- 20019204 TI - Exchange transfusion and intravenous immunoglobulin use in the UK. PMID- 20019205 TI - Need for consensus in interpreting coagulation profile in preterm neonates. PMID- 20019206 TI - Timing of repeat elective caesarean delivery and neonatal respiratory outcomes. PMID- 20019207 TI - The polypill concept. PMID- 20019208 TI - Pascal's Wager, infective endocarditis and the "no-lose" philosophy in medicine. AB - Doctors and dentists have traditionally used antibiotic prophylaxis in certain patient groups in order to prevent infective endocarditis (IE). New guidelines, however, suggest that the risk to patients from using antibiotics is higher than the risk from IE. This paper analyses the relative risks of prescribing and not prescribing antibiotic prophylaxis against the background of Pascal's Wager, the infamous assertion that it is better to believe in God regardless of evidence, because of the prospective benefits should He exist. Many doctors seem to believe the parallel proposition that it is better to prescribe antibiotics, regardless of evidence, because of the prospective benefit conferred upon the patient. This has been called the "no lose philosophy" in medicine: better safe than sorry, even if the evidence inconveniently suggests that following this mantra is potentially more likely to result in sorry than safe. It transpires that, just as Pascal's Wager fails to convince because of a lack of evidence to support it and the costs incurred by trying to believe, so the "belts and braces" approach of prescribing antibiotic prophylaxis is unjustifiable given the actual evidence of potential risk and benefit to the patient. Ultimately, there is no no-lose if your clinical decisions, like Pascal's Wager, are based on faith rather than evidence. PMID- 20019209 TI - "Barred" mitral valve. PMID- 20019210 TI - Management of heart failure with preserved ejection fraction: back to the drawing board? PMID- 20019211 TI - The impact of prehospital thrombolytic treatment on re-infarction rates. PMID- 20019212 TI - Triple antithrombotic management after stent implantation: when and how? PMID- 20019213 TI - Echocardiographic follow-up after heart valve replacement. PMID- 20019214 TI - Renal disease and acute coronary syndrome. PMID- 20019215 TI - Why an Australian editor for JNNP? PMID- 20019216 TI - Stop using the Ashworth scale for the assessment of spasticity. PMID- 20019217 TI - Cerebral spinal fluid abnormalities in patients with paraneoplastic syndromes of the nervous system. PMID- 20019218 TI - "Benign" and "normal appearing": it's in the eye of the beholder. PMID- 20019219 TI - The role of DAT-SPECT in movement disorders. AB - Dopamine transporter (DAT) imaging is a sensitive method to detect presynaptic dopamine neuronal dysfunction, which is a hallmark of neurodegenerative parkinsonism. DAT imaging can therefore assist the differentiation between conditions with and without presynaptic dopaminergic deficit. Diagnosis of Parkinson disease or tremor disorders can be achieved with high degrees of accuracy in cases with full expression of classical clinical features; however, diagnosis can be difficult, since there is a substantial clinical overlap especially in monosymptomatic tremor (dystonic tremor, essential tremor, Parkinson tremor). The use of DAT-SPECT can prove or excludes with high sensitivity nigrostriatal dysfunction in those cases and facilitates early and accurate diagnosis. Furthermore, a normal DAT-SPECT is helpful in supporting a diagnosis of drug-induced-, psychogenic- and vascular parkinsonism by excluding underlying true nigrostriatal dysfunction. This review addresses the value of DAT SPECT and its impact on diagnostic accuracy in movement disorders presenting with tremor and/or parkinsonism. PMID- 20019220 TI - Neurological picture. Cerebral proliferative angiopathy associated with haemangioma of the face and tongue. PMID- 20019221 TI - Neurological picture. Cortical oedema: a link between delusional misidentification syndromes and hemiplegic migraine. PMID- 20019222 TI - A multicentre study on the clinical utility of post-traumatic amnesia duration in predicting global outcome after moderate-severe traumatic brain injury. AB - BACKGROUND: Past research shows that post-traumatic amnesia (PTA) duration is a particularly robust traumatic brain injury (TBI) outcome predictor, but low specificity limits its clinical utility. OBJECTIVES: The current study assessed the relationship between PTA duration and probability thresholds for Glasgow Outcome Scale (GOS) levels. METHODS: Data were prospectively collected in this multicentre observational study. The cohort was a consecutive sample of rehabilitation patients enrolled in the National Institute on Disability and Rehabilitation Research funded TBI Model Systems (n = 1332) that had documented finite PTA duration greater than 24 h, and 1-year and 2-year GOS. RESULTS: The cohort had proportionally more Good Recovery (44% vs 39%) and less Severe Disability (19% vs 23%) at year 2 than at year 1. Longer PTA resulted in an incremental decline in probability of Good Recovery and a corresponding increase in probability of Severe Disability. When PTA ended within 4 weeks, Severe Disability was unlikely (<15% chance) at year 1, and Good Recovery was the most likely GOS at year 2. When PTA lasted beyond 8 weeks, Good Recovery was highly unlikely (<10% chance) at year 1, and Severe Disability was equal to or more likely than Moderate Disability at year 2. CONCLUSIONS: Two PTA durations, 4 weeks and 8 weeks, emerged as particularly salient GOS probability thresholds that may aid prognostication after TBI. PMID- 20019223 TI - Progressive myoclonic epilepsy as an adult-onset manifestation of Leigh syndrome due to m.14487T>C. AB - BACKGROUND: m.14487T>C, a missense mutation (p.M63V) affecting the ND6 subunit of complex I of the mitochondrial respiratory chain, has been reported in isolated childhood cases with Leigh syndrome (LS) and progressive dystonia. Adult-onset phenotypes have not been reported. OBJECTIVES: To determine the clinical neurological spectrum and associated mutation loads in an extended m.14487T>C family. METHODS: A genotype-phenotype correlation study of a Belgian five generation family with 12 affected family members segregating m.14487T>C was carried out. Clinical and mutation load data were available for nine family members. Biochemical analysis of the respiratory chain was performed in three muscle biopsies. RESULTS: Heteroplasmic m.14487T>C levels (36-52% in leucocytes, 97-99% in muscle) were found in patients with progressive myoclonic epilepsy (PME) and dystonia or progressive hypokinetic-rigid syndrome. Patients with infantile LS were homoplasmic (99-100% in leucocytes, 100% in muscle). We found lower mutation loads (between 8 and 35% in blood) in adult patients with clinical features including migraine with aura, Leber hereditary optic neuropathy, sensorineural hearing loss and diabetes mellitus type 2. Despite homoplasmic mutation loads, complex I catalytic activity was only moderately decreased in muscle tissue. INTERPRETATION: m.14487T>C resulted in a broad spectrum of phenotypes in our family. Depending on the mutation load, it caused severe encephalopathies ranging from infantile LS to adult-onset PME with dystonia. This is the first report of PME as an important neurological manifestation of an isolated mitochondrial complex I defect. PMID- 20019224 TI - Phenotypic spectrum of hereditary neuralgic amyotrophy caused by the SEPT9 R88W mutation. AB - BACKGROUND: Hereditary neuralgic amyotrophy (HNA), also known as hereditary brachial plexus neuropathy, has phenotypic and genetic heterogeneity. Mutations in the septin 9 (SEPT9) gene were recently identified in some HNA patients. The phenotypic spectrum of HNA caused by SEPT9 mutations is not well known. OBJECTIVE: To characterise the phenotype of a large family of HNA patients with the SEPT9 R88W mutation. METHODS: We report clinical, electrophysiological, neuroimaging and genetic findings of six HNA patients from a Japanese family. RESULTS: All 17 neuropathic episodes identified were selectively and asymmetrically distributed in the upper-limb nerves. Severe pain was an initial symptom in 16 episodes (94%). Motor weakness occurred in 15 (88%) and sensory signs in 10 (59%). A minor dysmorphism, hypotelorism, was seen in all. Nerve conduction studies revealed focal demyelination as well as prominent axonal degeneration changes. Needle electromyography revealed chronic neurogenic patterns only in the upper limbs. An MRI study showed a gadolinium-enhanced brachial plexus. The missense mutation c.262C>T; p.R88W was found in exon 2 of SEPT9 in all patients. CONCLUSIONS: The SEPT9 R88W mutation in this family causes selective involvement of the brachial plexus and upper-limb nerves. Wider and more universal recognition of clinical hallmarks and genetic counselling are of diagnostic importance for HNA caused by the SEPT9 mutation. PMID- 20019225 TI - Cortical excitability in hereditary motor neuronopathy with pyramidal signs: comparison with ALS. AB - BACKGROUND: Distal hereditary motor neuronopathy with pyramidal features (dHMNP) is a hereditary neurodegenerative disorder characterised by the presence of upper and lower motor neuron signs. The pathophysiological mechanisms underlying these clinical findings remain elusive. Given that cortical hyperexcitability appears to underlie neurodegeneration in amyotrophic lateral sclerosis (ALS), a disorder that may clinically resemble dHMNP, the present study applied novel cortical excitability studies to further investigate the pathophysiological mechanisms in dHMNP. METHODS: Threshold tracking transcranial magnetic stimulation (TMS) studies were undertaken using a 90 mm circular coil. Peripheral nerve excitability was performed by stimulating the median nerve at the wrist, with recording made over the abductor pollicis brevis muscle. Studies were undertaken in six dHMNP and 52 ALS patients, and compared with 55 normal controls. RESULTS: Central motor conduction time (CMCT) was significantly prolonged in dHMNP (dHMNP 7.7 (SEM 0.7) ms; ALS 4.9 (0.3) ms; controls 5.1 (0.2) ms, p<0.01). Short interval intacortical inhibition (SICI) was significantly reduced in ALS patients (0.8 (0.8)%) when compared with dHMNP (6.4 (0.7)%, p<0.0001) and controls (8.6 (1.1)%, p<0.0001). Reduction in SICI was accompanied by significant increases in the magnetic stimulus-response curve gradient and intracortical facilitation, and reduction in cortical silent period duration in ALS, while all these parameters of cortical excitability were normal in dHMNP. CONCLUSIONS: The present study has established a prolonged CMCT and normal cortical excitability in dHMNP, thereby providing further support for the hypothesis that cortical hyperexcitability underlies neurodegeneration in ALS. PMID- 20019226 TI - Late-onset cervicoscapular muscle atrophy and weakness after radiotherapy for Hodgkin disease: a case series. AB - Patients with cervical or mediastinal Hodgkin disease (HD) classically underwent chemotherapy plus extended-field radiation therapy. We report six patients who gradually developed severe atrophy and weakness of cervical paraspinal and shoulder girdle muscles 5-30 years after mantle irradiation for HD. Although clinical presentation was uniform, including a dropped head syndrome, electrophysiological and pathological findings were rather heterogeneous. Either neurogenic or myogenic processes may be involved and sometimes combined. We discuss the pathophysiological mechanisms underlying these cervicoscapular motor complications of mantle irradiation in HD. PMID- 20019227 TI - Single versus bilateral external ventricular drainage for intraventricular fibrinolysis in severe ventricular haemorrhage. AB - BACKGROUND: Intraventricular fibrinolysis (IVF) through bilateral external ventricular drains (EVD) may provide better access of the thrombolytic agent to the intraventricular clot, potentially influencing clot clearance and outcome. METHODS: Patients with spontaneous ganglionic intracerebral haemorrhage (ICH)<40 cm(3) and intraventricular haemorrhage (IVH) with acute hydrocephalus have been treated with IVF. The decision for placement of one or two EVDs has been left to the discretion of the treating physician. CT volumetry, the effects on cerebrospinal fluid (CSF) circulation and outcome at 3 months have been analysed for patients with one (group I, n = 13) or two EVDs (group II, n = 14). RESULTS: No difference was found in clot resolution between the two groups (clot half life 2.1 (SD 1.2) vs 2.4 (1.3) days). A separate analysis of the third and fourth ventricle clearance was similar (1.6 (0.6) versus 1.8 (0.8) days), indicating no difference in reconstitution of CSF circulation. A trend towards a longer EVD duration and higher infection rate was found in the bilateral EVD group. No difference was found in outcome at 3 months. CONCLUSIONS: Our results do not support the use of bilateral EVDs for IVF in patients with severe IVH. PMID- 20019228 TI - Neuromyelitis optica-IgG (aquaporin-4) autoantibodies in immune mediated optic neuritis. AB - The clinical course of immune mediated optic neuritis (ON) will depend on the specific underlying inflammatory disease. These disorders have traditionally been classified according to clinical and MRI findings. Aquaporin-4 (AQP4) autoantibodies (neuromyelitis optica-IgG (NMO-IgG)) may have diagnostic and prognostic value in patients who present with isolated ON. In this prospective study, NMO-IgG was evaluated in 114 patients with ON in the following contexts: neuromyelitis optica (NMO), multiple sclerosis (MSON), chronic relapsing inflammatory ON (CRION), relapsing isolated ON (RION) and single isolated ON (SION). The proportion seropositive was 56% for NMO (n = 9), 0% for MSON (n = 28) and 5% for the remaining diagnostic categories (CRION (n = 19), RION (n = 17) and SION (n = 41)). Testing for NMO-IgG in patients with recurrent or severe ON who lack convincing evidence of MS may identify patients who would benefit from immunosuppression rather than MS directed immunomodulatory therapies. PMID- 20019229 TI - Variant Creutzfeldt-Jakob disease: the first confirmed case from Portugal shows early onset, long duration and unusual pathology. AB - We present clinical and autopsy findings in the first case of variant Creutzfeldt Jakob disease diagnosed and confirmed in Portugal. Onset was at 11 years, the earliest onset reported, and the course (32 months) relatively long. Western blot showed protease resistant prion protein, mainly of type 4 (2B) isoform. The cerebral cortex revealed severe spongiform change with numerous amyloid plaques, which did not fit the definition of florid plaques. In the striatum, spongiform change was limited but the extracellular space was dilated. Other reports have found marked spongiform change in the striatum and little in the cortex. Massive neuronal loss, in excess of what has been described, was found in the thalamus and pontine grey. The cerebellum showed, as expected, severe loss of granule cells, moderate loss of Purkinje cells and marked immunopositivity for the prion protein. Differences between our findings and previous ones probably result from the patient's long survival. PMID- 20019230 TI - Neurological picture. Acute Wernicke's encephalopathy with hyperechogenic corpora mammillaria in brain sonography. PMID- 20019231 TI - Differences in risk factors according to the site of intracranial aneurysms. AB - BACKGROUND AND PURPOSE: Several risk factors for aneurysmal subarachnoid haemorrhage have been identified but it is not known whether some sites of aneurysms are linked to a specific risk factor. In a series of patients with aneurysmal subarachnoid haemorrhage, we compared risk factors according to the site of the ruptured aneurysm at the circle of Willis. METHODS: From our prospectively collected database of patients with aneurysmal subarachnoid haemorrhage admitted to our hospital between 2003 and 2007, we retrieved 304 patients with saccular aneurysms on the anterior communicating artery, middle cerebral artery, posterior communicating artery, basilar artery and vertebral artery. Risk factors (age, gender, smoking, no or excessive alcohol intake, hypertension and familial preponderance) were assessed per aneurysm location and compared with the anterior communicating artery as reference. We calculated odds ratios (ORs) and corresponding 95% CI. RESULTS: In comparison with aneurysms at the anterior communicating artery, those at the middle cerebral artery were less associated with age >55 years (OR 0.4; 95% CI 0.2 to 0.8), those at the posterior communicating artery were less associated with male gender (OR 0.4; 95% CI 0.2 to 0.9) and those at the basilar artery were more associated with no alcohol consumption (OR 5.8; 95% CI 1.1 to 29.9). CONCLUSION: Risk factors differ according to the site of aneurysm. This heterogeneity should be kept in mind in studies on the aetiology of aneurysms, such as genetic studies. PMID- 20019232 TI - No effect of donepezil on striatal dopamine release in mild to moderate Alzheimer's disease. AB - OBJECTIVE: Work in experimental animals suggests that an interaction with dopaminergic networks might explain some of the therapeutic effects of the cholinesterase inhibitor class of drugs. This study aimed to test whether acute (single 5 mg) or 4-8 weeks (5-10 mg) of treatment with oral donepezil would elicit measurable striatal dopamine (DA) release in patients with mild to moderate Alzheimer's disease. A second aim was to establish whether any increase in DA levels would be associated with improvements in cognitive and motor function. METHODS: Percentage change in [(11)C]-raclopride (RAC) binding potential (BP(ND)) between baseline and treatment conditions was used to provide a measure of DA release. Repeated measures ANOVA was used to determine the effect of treatment on [(11)C]-RAC BP(ND) and neuropsychological test performance. RESULTS: Contrary to our prediction there was no significant change in [(11)C]- RAC BP(ND) after acute or a mean of 6 weeks (range 4-12) of treatment with donepezil. Although motor speed (finger tapping) improved following 4-12 weeks of treatment with donepezil (F(1,19) = 8.7, p = 0.009), this was not associated with the degree of change in [(11)C]-RAC BP(ND). CONCLUSIONS: Our findings provide no evidence that striatal DA levels are altered during the first 3 months of donepezil treatment. However, we cannot rule out the possibility that extrastriatal effects may be occurring. PMID- 20019233 TI - "Phantom" restless legs syndrome. PMID- 20019234 TI - Obstructive sleep apnoea syndrome: a possible cause of symptomatic hyperckaemia. PMID- 20019235 TI - CSF biomarker utilisation and ethical considerations of biomarker assisted diagnosis and research in dementia: perspectives from within the European Alzheimer's Disease Consortium (EADC). PMID- 20019236 TI - Acquired rippling muscle disease in association with myasthenia gravis. PMID- 20019239 TI - Glucose restriction can extend normal cell lifespan and impair precancerous cell growth through epigenetic control of hTERT and p16 expression. AB - Cancer cells metabolize glucose at elevated rates and have a higher sensitivity to glucose reduction. However, the precise molecular mechanisms leading to different responses to glucose restriction between normal and cancer cells are not fully understood. We analyzed normal WI-38 and immortalized WI-38/S fetal lung fibroblasts and found that glucose restriction resulted in growth inhibition and apoptosis in WI-38/S cells, whereas it induced lifespan extension in WI-38 cells. Moreover, in WI-38/S cells glucose restriction decreased expression of hTERT (human telomerase reverse transcriptase) and increased expression of p16(INK4a). Opposite effects were found in the gene expression of hTERT and p16 in WI-38 cells in response to glucose restriction. The altered gene expression was partly due to glucose restriction-induced DNA methylation changes and chromatin remodeling of the hTERT and p16 promoters in normal and immortalized WI 38 cells. Furthermore, glucose restriction resulted in altered hTERT and p16 expression in response to epigenetic regulators in WI-38 rather than WI-38/S cells, suggesting that energy stress-induced differential epigenetic regulation may lead to different cellular fates in normal and precancerous cells. Collectively, these results provide new insights into the epigenetic mechanisms of a nutrient control strategy that may contribute to cancer therapy as well as antiaging approaches. PMID- 20019240 TI - Differential levels of transcription of p53-regulated genes by the arginine/proline polymorphism: p53 with arginine at codon 72 favors apoptosis. AB - Human populations contain a functional coding polymorphism (codon 72) in the p53 gene. To explore whether this polymorphism alters the transcriptional pattern of p53-regulated genes, the human isogenic cell lines harboring p53 with either the proline or arginine at codon 72 were employed to activate p53-mediated transcription. Thirty-four p53-regulated genes were assayed for their increased levels of mRNA using quantitative real-time PCR. The largest difference between p53-arginine and p53-proline was found with the PERP gene involved in cell-cell adhesion and apoptosis. The most common set of genes that are transcribed better by the p53-arginine protein than the p53-proline protein was found in the apoptotic function (DR-4, NOXA, PUMA, and PIG-3). LIF, a cytokine that is required for optimal reproductive function, was produced at 2x higher levels by the p53-arginine than the p53-proline allele. The genes that induced their mRNAs at the highest levels compared to the baseline tended to be synthesized better by the p53-arginine protein than the p53-proline protein. These molecular studies may help to explain the complicated associations observed between this polymorphism and the incidence of some cancers, the longevity of some populations, and the fecundity of different groups. PMID- 20019241 TI - The highly virulent variola and monkeypox viruses express secreted inhibitors of type I interferon. AB - Variola virus (VARV) caused smallpox, one of the most devastating human diseases and the first to be eradicated, but its deliberate release represents a dangerous threat. Virulent orthopoxviruses infecting humans, such as monkeypox virus (MPXV), could fill the niche left by smallpox eradication and the cessation of vaccination. However, immunomodulatory activities and virulence determinants of VARV and MPXV remain largely unexplored. We report the molecular characterization of the VARV- and MPXV-secreted type I interferon-binding proteins, which interact with the cell surface after secretion and prevent type I interferon responses. The proteins expressed in the baculovirus system have been purified, and their interferon-binding properties characterized by surface plasmon resonance. The ability of these proteins to inhibit a broad range of interferons was investigated to identify potential adaptation to the human immune system. Furthermore, we demonstrate by Western blot and activity assays the expression of the type I interferon inhibitor during VARV and MPXV infections. These findings are relevant for the design of new vaccines and therapeutics to smallpox and emergent virulent orthopoxviruses because the type I interferon-binding protein is a major virulence factor in animal models, vaccination with this protein induces protective immunity, and its neutralization prevents disease progression. PMID- 20019242 TI - Cardiomyocyte VEGFR-1 activation by VEGF-B induces compensatory hypertrophy and preserves cardiac function after myocardial infarction. AB - Mounting evidence indicates that the function of members of the vascular endothelial growth factor (VEGF) family extends beyond blood vessel formation. Here, we show that the prolonged intramyocardial expression of VEGF-A(165) and VEGF-B(167) on adeno-associated virus-mediated gene delivery determined a marked improvement in cardiac function after myocardial infarction in rats, by promoting cardiac contractility, preserving viable cardiac tissue, and preventing remodeling of the left ventricle (LV) over time. Consistent with this functional outcome, animals treated with both factors showed diminished fibrosis and increased contractile myocardium, which were more pronounced after expression of the selective VEGF receptor-1 (VEGFR-1) ligand VEGF-B, in the absence of significant induction of angiogenesis. We found that cardiomyocytes expressed VEGFR-1, VEGFR-2, and neuropilin-1 and that, in particular, VEGFR-1 was specifically up-regulated in hypoxia and on exposure to oxidative stress. VEGF-B exerted powerful antiapoptotic effect in both cultured cardiomyocytes and after myocardial infarction in vivo. Finally, VEGFR-1 activation by VEGF-B was found to elicit a peculiar gene expression profile proper of the compensatory, hypertrophic response, consisting in activation of alphaMHC and repression of betaMHC and skeletal alpha-actin, and an increase in SERCA2a, RYR, PGC1alpha, and cardiac natriuretic peptide transcripts, both in cultured cardiomyocytes and in infarcted hearts. The finding that VEGFR-1 activation by VEGF-B prevents loss of cardiac mass and promotes maintenance of cardiac contractility over time has obvious therapeutic implications. PMID- 20019243 TI - Mechanical breaking of microtubules in axons during dynamic stretch injury underlies delayed elasticity, microtubule disassembly, and axon degeneration. AB - Little is known about which components of the axonal cytoskeleton might break during rapid mechanical deformation, such as occurs in traumatic brain injury. Here, we micropatterned neuronal cell cultures on silicone membranes to induce dynamic stretch exclusively of axon fascicles. After stretch, undulating distortions formed along the axons that gradually relaxed back to a straight orientation, demonstrating a delayed elastic response. Subsequently, swellings developed, leading to degeneration of almost all axons by 24 h. Stabilizing the microtubules with taxol maintained the undulating geometry after injury but greatly reduced axon degeneration. Conversely, destabilizing microtubules with nocodazole prevented undulations but greatly increased the rate of axon loss. Ultrastructural analyses of axons postinjury revealed immediate breakage and buckling of microtubules in axon undulations and progressive loss of microtubules. Collectively, these data suggest that dynamic stretch of axons induces direct mechanical failure at specific points along microtubules. This microtubule disorganization impedes normal relaxation of the axons, resulting in undulations. However, this physical damage also triggers progressive disassembly of the microtubules around the breakage points. While the disintegration of microtubules allows delayed recovery of the "normal" straight axon morphology, it comes at a great cost by interrupting axonal transport, leading to axonal swelling and degeneration. PMID- 20019244 TI - Stable expression, activity, and inducibility of cytochromes P450 in differentiated HepaRG cells. AB - HepaRG cells possess the unique property to differentiate in vitro and to express various functions of mature hepatocytes, including the major cytochromes P450 (P450s). In the present study, we carefully analyzed mRNA expression and activity of the major P450s and their responsiveness to three prototypical inducers, phenobarbital, rifampicin, and omeprazole, in differentiated HepaRG cell cultures over a 4-week period after low and high seeding. Only minor differences were observed in P450 activities when measured by two cocktails of probe substrates, probably related to the choice and/or concentration of substrates. Similar results were obtained from the two cell seeding conditions. Expression and activities of several P450s were dimethyl sulfoxide-dependent. However, basal P450 expression and activities as well as their responsiveness to the prototypical inducers were well maintained over the 4-week period, and a good correlation was observed between transcript levels and corresponding activities. Thus, CYP1A2, CYP2B6, and CYP3A4 were found to accurately respond to their respective prototypical inducers, i.e., omeprazole, phenobarbital, and rifampicin. Likewise, basal expression of several phase II enzymes, transporters, and nuclear receptors, and response to inducers were also well preserved. More genes were found to be induced in HepaRG cells than in primary human hepatocytes, and no marked variation was noticed between the different passages. Taken together, these data support the conclusion that HepaRG cells represent a promising surrogate to primary human hepatocytes for xenobiotic metabolism and toxicity studies. PMID- 20019246 TI - Editorial expression of concern. PMID- 20019245 TI - Metabolism and excretion of the novel bioreductive prodrug PR-104 in mice, rats, dogs, and humans. AB - PR-104 is the phosphate ester of a 3,5-dinitrobenzamide nitrogen mustard (PR 104A) that is reduced to active hydroxylamine and amine metabolites by reductases in tumors. In this study, we evaluate the excretion of [(3)H]PR-104 in mice and determine its metabolite profile in mice, rats, dogs, and humans after a single intravenous dose. Total radioactivity was rapidly and quantitatively excreted in mice, with cumulative excretion of 46% in urine and 50% in feces. The major urinary metabolites in mice were products from oxidative N-dealkylation and/or glutathione conjugation of the nitrogen mustard moiety, including subsequent mercapturic acid pathway metabolites. A similar metabolite profile was seen in mouse bile, mouse plasma, and rat urine and plasma. Dogs and humans also showed extensive thiol conjugation but little evidence of N-dealkylation. Humans, like rodents, showed appreciable reduced metabolites in plasma, but concentrations of the cytotoxic amine metabolite (PR-104M) were higher in mice than humans. The most conspicuous difference in metabolite profile was the much more extensive O beta-glucuronidation of PR-104A in dogs and humans than in rodents. The structure of the O-beta-glucuronide (PR-104G) was confirmed by independent synthesis. Its urinary excretion was responsible for 13 +/- 2% of total dose in humans but only 0.8 +/- 0.1% in mice. Based on these metabolite profiles, biotransformation of PR 104 in rodents is markedly different from that in humans, suggesting that rodents may not be appropriate for modeling human biotransformation and toxicology of PR 104. PMID- 20019250 TI - The breakthroughs of 2009. PMID- 20019248 TI - Spontaneous and x-ray-triggered crystallization at long range in self-assembling filament networks. AB - We report here crystallization at long range in networks of like-charge supramolecular peptide filaments mediated by repulsive forces. The crystallization is spontaneous beyond a given concentration of the molecules that form the filaments but can be triggered by x-rays at lower concentrations. The crystalline domains formed by x-ray irradiation, with interfilament separations of up to 320 angstroms, can be stable for hours after the beam is turned off, and ions that screen charges on the filaments suppress ordering. We hypothesize that the stability of crystalline domains emerges from a balance of repulsive tensions linked to native or x-ray-induced charges and the mechanical compressive entrapment of filaments within a network. Similar phenomena may occur naturally in the cytoskeleton of cells and, if induced externally in biological or artificial systems, lead to possible biomedical and lithographic functions. PMID- 20019247 TI - p53 controls radiation-induced gastrointestinal syndrome in mice independent of apoptosis. AB - Acute exposure to ionizing radiation can cause lethal damage to the gastrointestinal (GI) tract, a condition called the GI syndrome. Whether the target cells affected by radiation to cause the GI syndrome are derived from the epithelium or endothelium and whether the target cells die by apoptosis or other mechanisms are controversial issues. Studying mouse models, we found that selective deletion of the proapoptotic genes Bak1 and Bax from the GI epithelium or from endothelial cells did not protect mice from developing the GI syndrome after sub-total-body gamma irradiation. In contrast, selective deletion of p53 from the GI epithelium, but not from endothelial cells, sensitized irradiated mice to the GI syndrome. Transgenic mice overexpressing p53 in all tissues were protected from the GI syndrome after irradiation. These results suggest that the GI syndrome is caused by the death of GI epithelial cells and that these epithelial cells die by a mechanism that is regulated by p53 but independent of apoptosis. PMID- 20019249 TI - Objective confirmation of subjective measures of human well-being: evidence from the U.S.A. AB - A huge research literature, across the behavioral and social sciences, uses information on individuals' subjective well-being. These are responses to questions--asked by survey interviewers or medical personnel--such as, "How happy do you feel on a scale from 1 to 4?" Yet there is little scientific evidence that such data are meaningful. This study examines a 2005-2008 Behavioral Risk Factor Surveillance System random sample of 1.3 million U.S. citizens. Life satisfaction in each U.S. state is measured. Across America, people's answers trace out the same pattern of quality of life as previously estimated, from solely nonsubjective data, in one branch of economics (so-called "compensating differentials" neoclassical theory, originally from Adam Smith). There is a state by-state match (r = 0.6, P < 0.001) between subjective and objective well-being. This result has some potential to help to unify disciplines. PMID- 20019251 TI - Video: Ardipithecus ramidus. PMID- 20019252 TI - Breakthrough of the year. Ardipithecus ramidus. PMID- 20019253 TI - Breakthrough of the year. The runners-up. PMID- 20019254 TI - Scorecard. Rating last year's areas to watch. PMID- 20019255 TI - Breakdown revisited. Trying to stay afloat. PMID- 20019256 TI - Breakthrough of the year. Areas to watch. PMID- 20019257 TI - Virus of the year. The novel H1N1 influenza. PMID- 20019258 TI - 2010 U.S. budget. Congress takes care of science in quiet finish to a busy year. PMID- 20019259 TI - Academic freedom. Terrorism charges against grad student raise questions. PMID- 20019262 TI - U.S. science policy. Chair of science panel to leave Congress. PMID- 20019260 TI - Scientific integrity. A dark tale behind two retractions. PMID- 20019263 TI - Evolution. Spineless fish and dark flies prove gene regulation crucial. PMID- 20019265 TI - France. Sarkozy's 'Grand Loan' bets that research will pay off. PMID- 20019266 TI - Geology. Peril in the Pamirs. PMID- 20019267 TI - Geology. Burdened by Soviet legacy, nations spar over water rights. PMID- 20019268 TI - Marine sciences. U.S. poised to adopt national ocean policy. PMID- 20019269 TI - Cancer research. Melanoma drug vindicates targeted approach. PMID- 20019270 TI - Genomics. Ecological genomics gets down to genes--and function. PMID- 20019273 TI - Weighing reward and punishment. PMID- 20019271 TI - Time for DNA disclosure. PMID- 20019274 TI - Education. The nation's report card: a vision of large-scale science assessment. PMID- 20019275 TI - Evolution. Sexual selection and Darwin's mystery of mysteries. PMID- 20019276 TI - Physics. Universal few-body binding. PMID- 20019277 TI - Psychology. Racial bias, unspoken but heard. PMID- 20019278 TI - Engineering. Enabling new missions for robotic aircraft. PMID- 20019279 TI - Computer science. Mining our reality. PMID- 20019280 TI - Regiodivergent ring opening of chiral aziridines. AB - Kinetic resolution of strained three-membered rings has proven broadly useful for the generation of enantiopure organic intermediates. Herein we demonstrate a complementary approach whereby a single catalyst transforms a racemic mixture of aziridines to a pair of regioisomeric products, each in good yield with exceptionally high enantioselectivity. Specifically, the dimeric yttrium salen catalyst accelerates the ring opening of aliphatic aziridines by trimethylsilylazide, inducing nucleophilic attack at the primary position of one enantiomer and the secondary position of the other. Both rate and selectivity are highly sensitive to the catalyst structure. PMID- 20019281 TI - Stepwise modification of a modular enhancer underlies adaptation in a Drosophila population. AB - The evolution of cis regulatory elements (enhancers) of developmentally regulated genes plays a large role in the evolution of animal morphology. However, the mutational path of enhancer evolution--the number, origin, effect, and order of mutations that alter enhancer function--has not been elucidated. Here, we localized a suite of substitutions in a modular enhancer of the ebony locus responsible for adaptive melanism in a Ugandan Drosophila population. We show that at least five mutations with varied effects arose recently from a combination of standing variation and new mutations and combined to create an allele of large phenotypic effect. We underscore how enhancers are distinct macromolecular entities, subject to fundamentally different, and generally more relaxed, functional constraints relative to protein sequences. PMID- 20019283 TI - Formation and survival of water vapor in the terrestrial planet-forming region. AB - Recent astronomical observations have revealed what may prove to be the ubiquity of water vapor during the early stages of planet formation. We present here a simple mechanism showing how water vapor forms in situ and is capable of shielding itself from molecule-destroying stellar radiation. The absorption of this radiation by water can control the thermodynamics of the terrestrial planet forming zone. Similar to Earth's ozone layer, which shelters the chemistry of life, the water layer protects other water molecules and allows for a rich organic chemistry. The total abundance of water vapor in the natal habitable zone is equal to that of several thousand oceans. PMID- 20019282 TI - Crystal structure of the eukaryotic strong inward-rectifier K+ channel Kir2.2 at 3.1 A resolution. AB - Inward-rectifier potassium (K+) channels conduct K+ ions most efficiently in one direction, into the cell. Kir2 channels control the resting membrane voltage in many electrically excitable cells, and heritable mutations cause periodic paralysis and cardiac arrhythmia. We present the crystal structure of Kir2.2 from chicken, which, excluding the unstructured amino and carboxyl termini, is 90% identical to human Kir2.2. Crystals containing rubidium (Rb+), strontium (Sr2+), and europium (Eu3+) reveal binding sites along the ion conduction pathway that are both conductive and inhibitory. The sites correlate with extensive electrophysiological data and provide a structural basis for understanding rectification. The channel's extracellular surface, with large structured turrets and an unusual selectivity filter entryway, might explain the relative insensitivity of eukaryotic inward rectifiers to toxins. These same surface features also suggest a possible approach to the development of inhibitory agents specific to each member of the inward-rectifier K+ channel family. PMID- 20019284 TI - Spatial organization of hominin activities at Gesher Benot Ya'aqov, Israel. AB - The spatial designation of discrete areas for different activities reflects formalized conceptualization of a living space. The results of spatial analyses of a Middle Pleistocene Acheulian archaeological horizon (about 750,000 years ago) at Gesher Benot Ya'aqov, Israel, indicate that hominins differentiated their activities (stone knapping, tool use, floral and faunal processing and consumption) across space. These were organized in two main areas, including multiple activities around a hearth. The diversity of human activities and the distinctive patterning with which they are organized implies advanced organizational skills of the Gesher Benot Ya'aqov hominins. PMID- 20019285 TI - Mozambican grass seed consumption during the Middle Stone Age. AB - The role of starchy plants in early hominin diets and when the culinary processing of starches began have been difficult to track archaeologically. Seed collecting is conventionally perceived to have been an irrelevant activity among the Pleistocene foragers of southern Africa, on the grounds of both technological difficulty in the processing of grains and the belief that roots, fruits, and nuts, not cereals, were the basis for subsistence for the past 100,000 years and further back in time. A large assemblage of starch granules has been retrieved from the surfaces of Middle Stone Age stone tools from Mozambique, showing that early Homo sapiens relied on grass seeds starting at least 105,000 years ago, including those of sorghum grasses. PMID- 20019286 TI - Experimental observations of stress-driven grain boundary migration. AB - In crystalline materials, plastic deformation occurs by the motion of dislocations, and the regions between individual crystallites, called grain boundaries, act as obstacles to dislocation motion. Grain boundaries are widely envisaged to be mechanically static structures, but this report outlines an experimental investigation of stress-driven grain boundary migration manifested as grain growth in nanocrystalline aluminum thin films. Specimens fabricated with specially designed stress and strain concentrators are used to uncover the relative importance of these parameters on grain growth. In contrast to traditional descriptions of grain boundaries as stationary obstacles to dislocation-based plasticity, the results of this study indicate that shear stresses drive grain boundaries to move in a manner consistent with recent molecular dynamics simulations and theoretical predictions of coupled grain boundary migration. PMID- 20019287 TI - Indirect punishment and generosity toward strangers. AB - Many people incur costs to reward strangers who have been kind to others. Theoretical and experimental evidence suggests that such "indirect rewarding" sustains cooperation between unrelated humans. Its emergence is surprising, because rewarders incur costs but receive no immediate benefits. It can prevail in the long run only if rewarders earn higher payoffs than "defectors" who ignore strangers' kindness. We provide experimental evidence regarding the payoffs received by individuals who employ these and other strategies, such as "indirect punishment," by imposing costs on unkind strangers. We find that if unkind strangers cannot be punished, defection earns most. If they can be punished, however, then indirect rewarding earns most. Indirect punishment plays this important role, even if it gives a low payoff and is rarely implemented. PMID- 20019288 TI - The subtle transmission of race bias via televised nonverbal behavior. AB - Compared with more explicit racial slurs and statements, biased facial expressions and body language may resist conscious identification and thus produce a hidden social influence. In four studies, we show that race biases can be subtly transmitted via televised nonverbal behavior. Characters on 11 popular television shows exhibited more negative nonverbal behavior toward black than toward status-matched white characters. Critically, exposure to prowhite (versus problack) nonverbal bias increased viewers' bias even though patterns of nonverbal behavior could not be consciously reported. These findings suggest that hidden patterns of televised nonverbal behavior influence bias among viewers. PMID- 20019290 TI - Screening for antibacterial inhibitors of the UDP-3-O-(R-3-hydroxymyristoyl)-N acetylglucosamine deacetylase (LpxC) using a high-throughput mass spectrometry assay. AB - A high-throughput mass spectrometry assay to measure the catalytic activity of UDP-3-O-(R-3-hydroxymyristoyl)-N-acetylglucosamine deacetylase, LpxC, is described. This reaction is essential in the biosynthesis of lipopolysaccharide (LPS) of gram-negative bacteria and is an attractive target for the development of new antibacterial agents. The assay uses the RapidFire mass spectrometry platform to measure the native LpxC substrate and the reaction product and thereby generates a ratiometric readout with minimal artifacts due to detection interference. The assay was robust in a high-throughput screen of a library of more than 700,000 compounds arrayed as orthogonal mixtures, with a median Z' factor of 0.74. Selected novel inhibitors from the screening campaign were confirmed as binding to LpxC by biophysical measurements using a thermal stability shift assay. Some inhibitors showed whole-cell antimicrobial activity against a sensitive strain of Escherichia coli with reduced LpxC activity (strain D22; minimum inhibitory concentrations ranging from 0.625-20 microg/mL). The results show that mass spectrometry-based screening is a valuable high-throughput screening tool for detecting inhibitors of enzymatic targets involving difficult to detect reactions. PMID- 20019291 TI - Integrating public health across sectors. PMID- 20019292 TI - George Washington Goler: the biggest crank and the best health officer in the United States. PMID- 20019293 TI - Nothing to work with but cleanliness: the training of African American traditional midwives in the South. PMID- 20019294 TI - HIV prevention interventions for black MSM. PMID- 20019295 TI - Nicotine anonymous may benefit nicotine-dependent individuals. PMID- 20019296 TI - Ethics-based public health policy? PMID- 20019297 TI - The ubiquity of energy-dense snack foods: a national multicity study. AB - OBJECTIVES: We assessed the availability and accessibility of energy-dense snacks in retail stores whose primary merchandise was not food and whether these varied by store type, region, or socioeconomic factors. METHODS: We conducted systematic observations of 1082 retail stores in 19 US cities and determined the availability and accessibility of 6 categories of energy-dense snack foods. RESULTS: Snack food was available in 41% of the stores; the most common forms were candy (33%), sweetened beverages (20%), and salty snacks (17%). These foods were often within arm's reach of the cash register queue. We observed snack foods in 96% of pharmacies, 94% of gasoline stations, 22% of furniture stores, 16% of apparel stores, and 29% to 65% of other types of stores. Availability varied somewhat by region but not by the racial or socioeconomic characteristics of nearby census tracts. CONCLUSIONS: Energy-dense snack foods and beverages, implicated as contributors to the obesity epidemic, are widely available in retail stores whose primary business is not food. The ubiquity of these products may contribute to excess energy consumption in the United States. PMID- 20019298 TI - Ancillary care in community-based public health intervention research. AB - Community-based public health intervention research in developing countries typically takes place not in clinics but in people's homes and other living spaces. Research subjects and their communities may lack adequate nutrition, clean water, sanitation, and basic preventive and therapeutic services. Researchers often encounter unmet health needs in their interactions with individual subjects and need ethical guidelines to help them decide how to respond. To what extent do researchers have an ethical obligation to provide ancillary care-health care beyond what is necessary to ensure scientific validity and subjects' safety? We discuss a case example from Nepal and propose a simple 2 step sequence of questions to aid decision making. PMID- 20019299 TI - Black-White health disparities in the United States and Chicago: a 15-year progress analysis. AB - OBJECTIVES: In an effort to examine national and Chicago, Illinois, progress in meeting the Healthy People 2010 goal of eliminating health disparities, we examined whether disparities between non-Hispanic Black and non-Hispanic White persons widened, narrowed, or stayed the same between 1990 and 2005. METHODS: We examined 15 health status indicators. We determined whether a disparity widened, narrowed, or remained unchanged between 1990 and 2005 by examining the percentage difference in rates between non-Hispanic Black and non-Hispanic White populations at both time points and at each location. We calculated P values to determine whether changes in percentage difference over time were statistically significant. RESULTS: Disparities between non-Hispanic Black and non-Hispanic White populations widened for 6 of 15 health status indicators examined for the United States (5 significantly), whereas in Chicago the majority of disparities widened (11 of 15, 5 significantly). CONCLUSIONS: Overall, progress toward meeting the Healthy People 2010 goal of eliminating health disparities in the United States and in Chicago remains bleak. With more than 15 years of time and effort spent at the national and local level to reduce disparities, the impact remains negligible. PMID- 20019300 TI - The relative health burden of selected social and behavioral risk factors in the United States: implications for policy. AB - OBJECTIVES: We sought to quantify the potential health impact of selected medical and nonmedical policy changes within the United States. METHODS: Using data from the 1997-2000 National Health Interview Surveys (linked to mortality data through 2002) and the 1996-2002 Medical Expenditure Panel Surveys, we calculated age specific health-related quality-of-life scores and mortality probabilities for 8 social and behavioral risk factors. We then used Markov models to estimate the quality-adjusted life years lost. RESULTS: Ranked quality-adjusted life years lost were income less than 200% of the poverty line versus 200% or greater (464 million; 95% confidence interval [CI]=368, 564); current-smoker versus never smoker (329 million; 95% CI=226, 382); body mass index 30 or higher versus 20 to less than 25 (205 million; 95% CI=159, 269); non-Hispanic Black versus non Hispanic White (120 million; 95% CI=83, 163); and less than 12 years of school relative to 12 or more (74 million; 95% CI=52, 101). Binge drinking, overweight, and health insurance have relatively less influence on population health. CONCLUSIONS: Poverty, smoking, and high-school dropouts impose the greatest burden of disease in the United States. PMID- 20019301 TI - The association of perceived abuse and discrimination after September 11, 2001, with psychological distress, level of happiness, and health status among Arab Americans. AB - OBJECTIVES: We assessed the prevalence of perceived abuse and discrimination among Arab American adults after September 11, 2001, and associations between abuse or discrimination and psychological distress, level of happiness, and health status. METHODS: We gathered data from a face-to-face survey administered in 2003 to a representative, population-based sample of Arab American adults residing in the greater Detroit area. RESULTS: Overall, 25% of the respondents reported post-September 11 personal or familial abuse, and 15% reported that they personally had a bad experience related to their ethnicity, with higher rates among Muslims than Christians. After adjustment for socioeconomic and demographic factors, perceived post-September 11 abuse was associated with higher levels of psychological distress, lower levels of happiness, and worse health status. Personal bad experiences related to ethnicity were associated with increased psychological distress and reduced happiness. Perceptions of not being respected within US society and greater reported effects of September 11 with respect to personal security and safety were associated with higher levels of psychological distress. CONCLUSIONS: Perceived post-September 11 abuse and discrimination were associated with increased psychological distress, reduced levels of happiness, and worse health status in our sample. Community-based, culturally sensitive partnerships should be established to assess and meet the health needs of Arab Americans. PMID- 20019303 TI - Prevalence and correlates of emotional, physical, sexual, and financial abuse and potential neglect in the United States: the National Elder Mistreatment Study. AB - OBJECTIVES: We estimated prevalence and assessed correlates of emotional, physical, sexual, and financial mistreatment and potential neglect (defined as an identified need for assistance that no one was actively addressing) of adults aged 60 years or older in a randomly selected national sample. METHODS: We compiled a representative sample by random digit dialing across geographic strata. We used computer-assisted telephone interviewing to standardize collection of demographic, risk factor, and mistreatment data. We subjected prevalence estimates and mistreatment correlates to logistic regression. RESULTS: We analyzed data from 5777 respondents. One-year prevalence was 4.6% for emotional abuse, 1.6% for physical abuse, 0.6% for sexual abuse, 5.1% for potential neglect, and 5.2% for current financial abuse by a family member. One in 10 respondents reported emotional, physical, or sexual mistreatment or potential neglect in the past year. The most consistent correlates of mistreatment across abuse types were low social support and previous traumatic event exposure. CONCLUSIONS: Our data showed that abuse of the elderly is prevalent. Addressing low social support with preventive interventions could have significant public health implications. PMID- 20019302 TI - Racial/ethnic differences in self-reported racism and its association with cancer related health behaviors. AB - OBJECTIVES: We used population-based survey data to estimate the prevalence of self-reported racism across racial/ethnic groups and to evaluate the association between self-reported racism and cancer-related health behaviors. METHODS: We used cross-sectional data from the 2003 California Health Interview Survey. Questions measured self-reported racism in general and in health care. The cancer risk behaviors we assessed were smoking, binge drinking, not walking, being overweight or obese, and not being up to date with screenings for breast, cervical, colorectal, and prostate cancers. Analyses included descriptive analyses and logistic regression. RESULTS: Prevalences of self-reported racism varied between and within aggregate racial/ethnic groups. In adjusted analyses, general racism was associated with smoking, binge drinking, and being overweight or obese; health care racism was associated with not being up to date with screening for prostate cancer. Associations varied across racial/ethnic groups. CONCLUSIONS: Associations between general racism and lifestyle behaviors suggest that racism is a potential stressor that may shape cancer-related health behaviors, and its impact may vary by race/ethnicity. PMID- 20019304 TI - Effects of repeated deployment to Iraq and Afghanistan on the health of New Jersey Army National Guard troops: implications for military readiness. AB - OBJECTIVES: We assessed the effects of prior military service in Iraq or Afghanistan on the health of New Jersey Army National Guard members preparing for deployment to Iraq. METHODS: We analyzed anonymous, self-administered predeployment surveys from 2543 National Guard members deployed to Iraq in 2008. We used bivariate and multivariate analyses to measure the effects of prior service in Afghanistan (Operation Enduring Freedom [OEF]) or Iraq (Operation Iraqi Freedom [OIF]) on mental and physical health. RESULTS: Nearly 25% of respondents reported at least 1 previous OEF or OIF deployment. Previously deployed soldiers were more than 3 times as likely as soldiers with no previous deployments to screen positive for posttraumatic stress disorder (adjusted odds ratio [AOR]=3.69; 95% confidence interval [CI]=2.59, 5.24) and major depression (AOR=3.07; 95% CI=1.81, 5.19), more than twice as likely to report chronic pain (AOR=2.20; 95% CI=1.78, 2.72) and more than 90% more likely to score below the general population norm on physical functioning (AOR=1.94; 95% CI=1.51, 2.48). CONCLUSIONS: Repeated OEF and OIF deployments may adversely affect the military readiness of New Jersey National Guard combat soldiers. PMID- 20019305 TI - LUCHAR: using computer technology to battle heart disease among Latinos. AB - Many promising technology-based programs designed to promote healthy behaviors such as physical activity and healthy eating have not been adapted for use with diverse communities, including Latino communities. We designed a community-based health kiosk program for English- and Spanish-speaking Latinos. Users receive personalized feedback on nutrition, physical activity, and smoking behaviors from computerized role models that guide them in establishing goals in 1 or more of these 3 areas. We found significant improvements in nutrition and physical activity among 245 Latino program users; however, no changes were observed with respect to smoking behaviors. The program shows promise for extending the reach of chronic disease prevention and self-management programs. PMID- 20019306 TI - The food industry and self-regulation: standards to promote success and to avoid public health failures. AB - Threatened by possible government regulation and critical public opinion, industries often undertake self-regulatory actions, issue statements of concern for public welfare, and assert that self-regulation is sufficient to protect the public. The food industry has made highly visible pledges to curtail children's food marketing, sell fewer unhealthy products in schools, and label foods in responsible ways. Ceding regulation to industry carries opportunities but is highly risky. In some industries (e.g., tobacco), self-regulation has been an abject failure, but in others (e.g., forestry and marine fisheries), it has been more successful. We examined food industry self-regulation in the context of other self-regulatory successes and failures and defined 8 standards that should be met if self-regulation is to be effective. PMID- 20019307 TI - Evaluating the impact of menu labeling on food choices and intake. AB - OBJECTIVES: We assessed the impact of restaurant menu calorie labels on food choices and intake. METHODS: Participants in a study dinner (n=303) were randomly assigned to either (1) a menu without calorie labels (no calorie labels), (2) a menu with calorie labels (calorie labels), or (3) a menu with calorie labels and a label stating the recommended daily caloric intake for an average adult (calorie labels plus information). Food choices and intake during and after the study dinner were measured. RESULTS: Participants in both calorie label conditions ordered fewer calories than those in the no calorie labels condition. When calorie label conditions were combined, that group consumed 14% fewer calories than the no calorie labels group. Individuals in the calorie labels condition consumed more calories after the study dinner than those in both other conditions. When calories consumed during and after the study dinner were combined, participants in the calorie labels plus information group consumed an average of 250 fewer calories than those in the other groups. CONCLUSIONS: Calorie labels on restaurant menus impacted food choices and intake; adding a recommended daily caloric requirement label increased this effect, suggesting menu label legislation should require such a label. Future research should evaluate menu labeling's impact on children's food choices and consumption. PMID- 20019308 TI - Timeliness of breast cancer diagnosis and initiation of treatment in the National Breast and Cervical Cancer Early Detection Program, 1996-2005. AB - OBJECTIVES: To determine the effects of program policy changes, we examined service delivery benchmarks for breast cancer screening in the National Breast and Cervical Cancer Early Detection Program (NBCCEDP). METHODS: We analyzed NBCCEDP data for women with abnormal mammogram or clinical breast examination (n=382 416) from which 23 701 cancers were diagnosed. We examined time to diagnosis and treatment for 2 time periods: 1996 to 2000 and 2001 to 2005. We compared median time for diagnostic, treatment initiation, and total intervals with the Kruskal-Wallis test. We calculated adjusted proportions (predicted marginals) with logistic regression to examine diagnosis and treatment within program benchmarks (2% of total brain volume (P=0.023) and altered mental status at presentation (P=0.005). CONCLUSIONS: Spontaneous childhood ICH was due mostly to vascular malformations. Acute surgical intervention was commonly performed. Although death was rare, 71% of survivors had persisting neurological deficits. Larger ICH volume and altered mental status predicted clinically significant disability. PMID- 20019326 TI - Noggin protects against ischemic brain injury in rodents. AB - BACKGROUND AND PURPOSE: Bone morphogenetic proteins and their receptors are expressed in adult brains, and their expression levels increase after cerebral ischemia. The brain also expresses an inhibitor of bone morphogenetic protein signaling, noggin, but the role of noggin in ischemic disease outcome has not been studied. METHODS: We used transgenic mice overexpressing noggin to assess whether inhibition of bone morphogenetic protein signaling affects ischemic injury responses after permanent middle cerebral artery occlusion. RESULTS: Transgenic mice overexpressing noggin mice had significantly smaller infarct volumes and lower motor deficits compared to wild-type mice. CD11b(+) and IBA1(+) microglia along with oligodendroglial progenitors were significantly increased in transgenic mice overexpressing noggin mice at 14 days after permanent middle cerebral artery occlusion. CONCLUSIONS: These results provide genetic evidence that overexpression of noggin reduces ischemic brain injury after permanent middle cerebral artery occlusion via enhanced activation of microglia and oligodendrogenesis. PMID- 20019327 TI - Brain-derived neurotrophic factor stimulates production of prostacyclin in cerebral arteries. AB - BACKGROUND AND PURPOSE: The role of brain-derived neurotrophic factor (BDNF) and its receptor, tropomyosin receptor kinase B, in control of cerebral circulation is poorly understood. The present study was designed to investigate the cerebral vascular effects of BDNF in vivo. METHODS: Replication incompetent adenovirus encoding either rat BDNF (AdBDNF) or green fluorescent protein was injected intracisternally into rabbits. Forty-eight hours later, animals were euthanized. Plasma and cerebrospinal fluid levels of BDNF were measured by enzyme-linked immunosorbent assay, vasomotor function of isolated basilar arteries was studied in organ chambers, protein expression in the basilar arteries was studied by Western blotting, prostanoid levels were measured by enzyme-linked immunosorbent assay, and cyclic adenosine 3',5'-monophosphate levels were measured by radioimmunoassay. RESULTS: The levels of BDNF in the cerebrospinal fluid were significantly elevated in AdBDNF-treated rabbits as compared with adenovirus encoding green fluorescent protein-treated rabbits (37+/-5 ng/mL versus 0.006+/ 0.003 ng/mL, respectively; P<0.05; n=14). Western blotting studies revealed that in basilar arteries, AdBDNF increased protein expression of prostacyclin synthase, whereas expression of endothelial nitric oxide synthase and phosphorylated (Ser 1177) endothelial nitric oxide synthase remained unchanged. During incubation with arachidonic acid (1 micromol/L), PGI(2) production and levels of cyclic adenosine 3',5'-monophosphate were significantly elevated only in AdBDNF-treated rabbit basilar arteries (P<0.05, n=6). Relaxations to acetylcholine (10(-9) to 10(-5) mol/L) and arachidonic acid (10(-9) to 10(-5) mol/L) were significantly potentiated in basilar arteries from rabbits injected with AdBDNF. Potentiation of relaxations to acetylcholine in AdBDNF-treated basilar arteries was inhibited by the nonselective cyclooxygenase inhibitor, indomethacin (10(-5) mol/L, P<0.05, n=6) and constitutive phospholipase A(2) inhibitor, AACOCF3 (2x10(-5) mol/L, P<0.05, n=5). CONCLUSIONS: Our results demonstrate that in cerebral arteries, BDNF-induced activation of tropomyosin receptor kinase B receptor signaling in vivo promotes prostacyclin biosynthesis. These findings provide novel mechanistic insight into the vascular protective effect of BDNF in cerebral circulation. PMID- 20019328 TI - Angiotensin II induces a region-specific hyperplasia of the ascending aorta through regulation of inhibitor of differentiation 3. AB - RATIONALE: Angiotensin II (Ang II) has diverse effects on smooth muscle cells (SMCs). The diversity of effects may relate to the regional location of this cell type. OBJECTIVE: The aim of this study was to define whether Ang II exerted divergent effects on smooth muscle cells in the aorta and determine the role of blood pressure and specific oxidant mechanisms. METHODS AND RESULTS: Ang II (1000 ng/kg per minute) infusion for 28 days into mice increased systolic blood pressure and promoted medial expansion of equivalent magnitude throughout the entire aorta. Both effects were ablated by angiotensin II type 1a (AT(1a)) receptor deficiency. Similar increases in systolic blood pressure by administration of norepinephrine promoted no changes in aortic medial thickness. Increased medial thickness was attributable to SMC expansion owing to hypertrophy in most aortic regions, with the exception of hyperplasia of the ascending aorta. Deficiency of the p47(phox) component of NADPH oxidase ablated Ang II-induced medial expansion in all aortic regions. Analysis of mRNA and protein throughout the aorta revealed a much higher abundance of the inhibitor of differentiation 3 (Id3) in the ascending aorta compared to all other regions. A functional role was demonstrated by Id3 deficiency inhibiting Ang II-induced SMC hyperplasia of the ascending aorta. CONCLUSIONS: In conclusion, Ang II promotes both aortic medial hypertrophy and hyperplasia in a region-specific manner via an oxidant mechanism. The ascending aortic hyperplasia is dependent on Id3. PMID- 20019329 TI - Fibulin-4 deficiency results in ascending aortic aneurysms: a potential link between abnormal smooth muscle cell phenotype and aneurysm progression. AB - RATIONALE: Loss of fibulin-4 during embryogenesis results in perinatal lethality because of aneurysm rupture, and defective elastic fiber assembly has been proposed as an underlying cause for the aneurysm phenotype. However, aneurysms are never seen in mice deficient for elastin, or for fibulin-5, which absence also leads to compromised elastic fibers. OBJECTIVE: We sought to determine the mechanism of aneurysm development in the absence of fibulin-4 and establish the role of fibulin-4 in aortic development. METHODS AND RESULTS: We generated germline and smooth muscle cell (SMC)-specific deletion of the fibulin-4 gene in mice (Fbln4(GKO) and Fbln4(SMKO), respectively). Fbln4(GKO) and Fbln4(SMKO) aortic walls fail to fully differentiate, exhibiting reduced expression of SM specific contractile genes and focal proliferation of SMCs accompanied by degenerative changes of the medial wall. Marked upregulation of extracellular signal-regulated kinase 1/2 signaling pathway was observed in the aneurysmal wall of Fbln4(GKO) and Fbln4(SMKO) mice and both mutants developed aneurysm predominantly in the ascending thoracic aorta. In vitro, Fbln4(GKO) SMCs exhibit an immature SMC phenotype with a marked reduction of SM-myosin heavy chain and increased proliferative capacity. CONCLUSIONS: The vascular phenotype in Fbln4 mutant mice is remarkably similar to a subset of human thoracic aortic aneurysms caused by mutations in SMC contractile genes. Our study provides a potential link between the intrinsic properties of SMCs and aneurysm progression in vivo and supports the dual role of fibulin-4 in the formation of elastic fibers as well as terminal differentiation and maturation of SMCs in the aortic wall. PMID- 20019330 TI - Chemoreceptor hypersensitivity, sympathetic excitation, and overexpression of ASIC and TASK channels before the onset of hypertension in SHR. AB - RATIONALE: Increased sympathetic nerve activity has been linked to the pathogenesis of hypertension in humans and animal models. Enhanced peripheral chemoreceptor sensitivity which increases sympathetic nerve activity has been observed in established hypertension but has not been identified as a possible mechanism for initiating an increase in sympathetic nerve activity before the onset of hypertension. OBJECTIVE: We tested this hypothesis by measuring the pH sensitivity of isolated carotid body glomus cells from young spontaneously hypertensive rats (SHR) before the onset of hypertension and their control normotensive Wistar-Kyoto (WKY) rats. METHODS AND RESULTS: We found a significant increase in the depolarizing effect of low pH in SHR versus WKY glomus cells which was caused by overexpression of 2 acid-sensing non-voltage-gated channels. One is the amiloride-sensitive acid-sensing sodium channel (ASIC3), which is activated by low pH and the other is the 2-pore domain acid-sensing K(+) channel (TASK1), which is inhibited by low pH and blocked by quinidine. Moreover, we found that the increase in sympathetic nerve activity in response to stimulation of chemoreceptors with sodium cyanide was markedly enhanced in the still normotensive young SHR compared to control WKY rats. CONCLUSIONS: Our results establish a novel molecular basis for increased chemotransduction that contributes to excessive sympathetic activity before the onset of hypertension. PMID- 20019331 TI - Hypoxia triggers subcellular compartmental redox signaling in vascular smooth muscle cells. AB - RATIONALE: Recent studies have implicated mitochondrial reactive oxygen species (ROS) in regulating hypoxic pulmonary vasoconstriction (HPV), but controversy exists regarding whether hypoxia increases or decreases ROS generation. OBJECTIVE: This study tested the hypothesis that hypoxia induces redox changes that differ among subcellular compartments in pulmonary (PASMCs) and systemic (SASMCs) smooth muscle cells. METHODS AND RESULTS: We used a novel, redox sensitive, ratiometric fluorescent protein sensor (RoGFP) to assess the effects of hypoxia on redox signaling in cultured PASMCs and SASMCs. Using genetic targeting sequences, RoGFP was expressed in the cytosol (Cyto-RoGFP), the mitochondrial matrix (Mito-RoGFP), or the mitochondrial intermembrane space (IMS RoGFP), allowing assessment of oxidant signaling in distinct intracellular compartments. Superfusion of PASMCs or SASMCs with hypoxic media increased oxidation of both Cyto-RoGFP and IMS-RoGFP. However, hypoxia decreased oxidation of Mito-RoGFP in both cell types. The hypoxia-induced oxidation of Cyto-RoGFP was attenuated through the overexpression of cytosolic catalase in PASMCs. CONCLUSIONS: These results indicate that hypoxia causes a decrease in nonspecific ROS generation in the matrix compartment, whereas it increases regulated ROS production in the IMS, which diffuses to the cytosol of both PASMCs and SASMCs. PMID- 20019332 TI - Effects of mechanical stress and carvedilol in lamin A/C-deficient dilated cardiomyopathy. AB - RATIONALE: Mutations in the LMNA gene, which encodes the nuclear lamina proteins lamin A and lamin C, are the most common cause of familial dilated cardiomyopathy (DCM). Mechanical stress-induced apoptosis has been proposed as the mechanism underpinning DCM in lamin A/C-deficient hearts, but supporting in vivo evidence has been lacking. OBJECTIVE: Our aim was to study interventions to modify mechanical stress in heterozygous Lmna knockout (Lmna(+/-)) mice. METHODS AND RESULTS: Cardiac structure and function were evaluated before and after exercise training, thoracic aortic constriction, and carvedilol treatment. Lmna(+/-) mice develop adult-onset DCM with relatively more severe disease in males. Lmna(+/-) cardiomyocytes show altered nuclear morphology and perinuclear desmin organization, with enhanced responses to hypo-osmotic stress indicative of cytoskeletal instability. Despite these structural defects that provide a template for mechanical stress-induced damage, young Lmna(+/-) mice subjected to 6 weeks of moderate or strenuous exercise training did not show induction of apoptosis or accelerated DCM. In contrast, regular moderate exercise attenuated DCM development in male Lmna(+/-) mice. Sustained pressure overload generated by thoracic aortic constriction depressed ventricular contraction in young wild-type and Lmna(+/-) mice with no sex or genotype differences in the time-course or severity of response. Treatment of male Lmna(+/-) mice from 12 to 40 weeks with the beta-blocker, carvedilol, prevented the dilatation and contractile dysfunction that was observed in placebo-treated mice. CONCLUSIONS: These data suggest that factors other than mechanical stress-induced apoptosis contribute to DCM and provide the first demonstration that regular moderate exercise and carvedilol can modify disease progression in lamin A/C-deficient hearts. PMID- 20019333 TI - Cartilage oligomeric matrix protein maintains the contractile phenotype of vascular smooth muscle cells by interacting with alpha(7)beta(1) integrin. AB - RATIONAL: Vascular smooth muscle cells (VSMCs) switching from a contractile/differentiated to a synthetic/dedifferentiated phenotype has an essential role in atherosclerosis, postangioplastic restenosis and hypertension. However, how normal VSMCs maintain the differentiated state is less understood. OBJECTIVE: We aimed to indentify the effect of cartilage oligomeric matrix protein (COMP), a normal vascular extracellular matrix, on modulation of VSMCs phenotype. METHODS AND RESULTS: We demonstrated that COMP was associated positively with the expression of VSMC differentiation marker genes during phenotype transition. Knockdown of COMP by small interfering (si)RNA favored dedifferentiation. Conversely, adenoviral overexpression of COMP markedly suppressed platelet-derived growth factor-BB-elicited VSMC dedifferentiation, characterized by altered VSMC morphology, actin fiber organization, focal adhesion assembly, and the expression of phenotype-dependent markers. Whereas alpha(7) integrin coimmunoprecipitated with COMP in normal rat VSMCs and vessels, neutralizing antibody or siRNA against alpha(7) integrin inhibited VSMC adhesion to COMP, which indicated that alpha(7)beta(1) integrin is a potential receptor for COMP. As well, blocking or interference by siRNA of alpha(7) integrin completely abolished the effect of COMP on conserving the contractile phenotype. In accordance, ectopic adenoviral overexpression of COMP greatly retarded VSMC phenotype switching, rescued contractility of carotid artery ring, and inhibited neointima formation in balloon-injured rats. CONCLUSIONS: Our data suggest that COMP is essential for maintaining a VSMC contractile phenotype and the protective effects of COMP are mainly mediated through interaction with alpha(7)beta(1) integrin. Investigations to identify the factors affecting the expression and integrity of COMP may provide a novel therapeutic target for vascular disorders. PMID- 20019335 TI - Re: ABO blood group and the risk of pancreatic cancer. PMID- 20019336 TI - Re: International guidelines for management of metastatic breast cancer: combination vs sequential single-agent chemotherapy. PMID- 20019334 TI - Overexpression of dimethylarginine dimethylaminohydrolase protects against cerebral vascular effects of hyperhomocysteinemia. AB - RATIONALE: Hyperhomocysteinemia is a cardiovascular risk factor that is associated with elevation of the nitric oxide synthase inhibitor asymmetrical dimethylarginine (ADMA). OBJECTIVE: Using mice transgenic for overexpression of the ADMA-hydrolyzing enzyme dimethylarginine dimethylaminohydrolase-1 (DDAH1), we tested the hypothesis that overexpression of DDAH1 protects from adverse structural and functional changes in cerebral arterioles in hyperhomocysteinemia. METHODS AND RESULTS: Hyperhomocysteinemia was induced in DDAH1 transgenic (DDAH1 Tg) mice and wild-type littermates using a high methionine/low folate (HM/LF) diet. Plasma total homocysteine was elevated approximately 3-fold in both wild type and DDAH1 Tg mice fed the HM/LF diet compared with the control diet (P<0.001). Plasma ADMA was approximately 40% lower in DDAH1 Tg mice compared with wild-type mice (P<0.001) irrespective of diet. Compared with the control diet, the HM/LF diet diminished endothelium-dependent dilation to 10 micromol/L acetylcholine in cerebral arterioles of both wild-type (12 + or - 2 versus 29 + or - 3%; P<0.001) and DDAH1 Tg (14 + or - 3 versus 28 + or - 2%; P<0.001) mice. Responses to 10 micromol/L papaverine, a direct smooth muscle dilator, were impaired with the HM/LF diet in wild-type mice (30 + or - 3 versus 45 + or - 5%; P<0.05) but not DDAH1 Tg mice (45 + or - 7 versus 48 + or - 6%). DDAH1 Tg mice also were protected from hypertrophy of cerebral arterioles (P<0.05) but not from accelerated carotid artery thrombosis induced by the HM/LF diet. CONCLUSIONS: Overexpression of DDAH1 protects from hyperhomocysteinemia-induced alterations in cerebral arteriolar structure and vascular muscle function. PMID- 20019337 TI - Serial OX40 engagement on CD4+ T cells and natural killer T cells causes allergic airway inflammation. AB - RATIONALE: OX40-OX40 ligand (OX40L) interactions have been proposed to support induction of allergic airway inflammation, which may be attributable to OX40 signaling in CD4(+) helper T cells for adaptive immune responses. However, a possible involvement of natural killer T (NKT) cells in the pathogenesis suggests that the underlying mechanisms are not yet fully elucidated. OBJECTIVES: We aimed to characterize the OX40-modulated cellular contribution to allergic airway inflammation in a mouse model of house dust mite (HDM) allergen exposure. METHODS: Mice were sensitized to HDM and, 3 weeks later, challenged with HDM on three consecutive days through the airways. Two days after the last exposure, bronchoalveolar lavage fluids and blood samples and lung tissues were evaluated for the airway inflammation. MEASUREMENTS AND MAIN RESULTS: The development of HDM-induced eosinophilic airway inflammation was dependent on OX40 of both CD4(+) T cells and NKT cells; OX40 engagement on CD4(+) T cells in the sensitization led to pulmonary OX40L augmentation after the allergen challenge, which stimulated pulmonary NKT cells through OX40 to provide the pathogenic cytokine milieu. This was ablated by OX40L blockade by inhalation of the neutralizing antibody during the challenge, suggesting the therapeutic potential of targeting pulmonary OX40 OX40L interactions. Moreover, OX40 expression in CD4(+) T cells, but not in NKT cells, was reciprocally regulated by the helper T cell type 1-skewing transcription factor Runx3. CONCLUSIONS: OX40 on not only CD4(+) T cells but also NKT cells is involved in allergic airway inflammation. Notably, pulmonary blockade of OX40 ligation on NKT cells has therapeutic implications. PMID- 20019338 TI - Murine airway luminal antituberculosis memory CD8 T cells by mucosal immunization are maintained via antigen-driven in situ proliferation, independent of peripheral T cell recruitment. AB - RATIONALE: The airway luminal memory CD8 T cells induced by respiratory mucosal immunization in a murine model have been found to be critical to antituberculosis immunity. However, the mechanisms of their maintenance on airway mucosal surface still remain poorly understood. OBJECTIVES: Using a model of adenovirus-based intranasal immunization we investigated the immune property and the mechanisms of maintenance of airway luminal CD8 T cells. METHODS: Immune properties of airway luminal Mycobacterium tuberculosis antigen-specific CD8 T cells were examined. Proliferation of airway luminal CD8 T cells was determined by in vivo T cell labeling techniques. The role of peripheral T cell recruitment in maintaining airway luminal CD8 T cells was investigated by blocking lymphocyte trafficking from lymphoid and peripheral tissues. The requirement of M. tuberculosis antigens for in situ T cell proliferation was evaluated using a T cell transfer approach. An airway M. tuberculosis challenge model was used to study the relationship between CD8 T cell-mediated protection and peripheral T cell recruitment. MEASUREMENTS AND MAIN RESULTS: Intranasal immunization leads to elicitation of persisting M. tuberculosis antigen-specific CD8 T cells in the airway lumen, which display an activated effector memory phenotype different from those in peripheral tissues. Airway luminal T cells continuously proliferate in an antigen dependent manner, and can be maintained even in the absence of peripheral T cell recruitment. The lungs equipped with such CD8 T cells are protected from airway M. tuberculosis challenge independent of both peripheral T cell supply and CD4 T cells. CONCLUSIONS: Vaccine-inducible airway luminal antituberculosis memory CD8 T cells are self-renewable in an antigen-dependent manner, and can be maintained independent of peripheral T cell supply. PMID- 20019339 TI - Reduced Th17 response in patients with tuberculosis correlates with IL-6R expression on CD4+ T Cells. AB - RATIONALE: Although it is well recognized that CD4(+) T cells and T helper (Th) 1 cytokines are critical in the cell-mediated response to Mycobacterium tuberculosis, it is also clear that this immunity alone is not enough. Understanding the roles of other T cell subsets and cytokines is essential for vaccine design and clinical immunotherapy against tuberculosis (TB). OBJECTIVES: To investigate the clinical significance and possible regulatory mechanism of Th17 responses in human TB. METHODS: The frequencies of IFN-gamma-, IL-4-, IL-17 , FoxP3- and IL-6 receptor (IL-6R)-expressing CD4(+) T cells in blood and/or pleural effusion samples of healthy donors, subjects with latent TB infection, and patients with active TB were analyzed by flow cytometry. Cytokines, transforming growth factor-beta and IL-6, in plasma and pleural fluid samples were determined by ELISA. MEASUREMENTS AND MAIN RESULTS: The frequency of Th17 cells in patients with active TB is significantly lower than those in healthy donors and individuals with latent TB infection. Correlation analysis showed that reduced Th17 responses observed in patients with active TB was significantly correlated with the decreased expression of IL-6R on CD4(+) T cells, but did not correlate with the concentrations of the cytokines, transforming growth factor beta and IL-6. Consistently; in vitro study showed that M. tuberculosis products inhibit the expression of IL-6R on CD4(+) T cells. CONCLUSIONS: Our results demonstrate that reduced Th17 responses were associated with the clinical outcome of M. tuberculosis infection. Suppression of Th17 response through down regulation of IL-6R expression may be an important mechanism in the development of active TB. PMID- 20019342 TI - Automated prediction of apnea and hypopnea, using a LAMSTAR artificial neural network. AB - RATIONALE: The prediction of individual episodes of apnea and hypopnea in people with obstructive sleep apnea syndrome has not been thoroughly investigated. Accurate prediction of these events could improve clinical management of this prevalent disease. OBJECTIVES: To evaluate the performance of a system developed to predict episodes of obstructive apnea and hypopnea in individuals with obstructive sleep apnea; to determine the most important signals for making accurate and reliable predictions. METHODS: We employed LArge Memory STorage And Retrieval (LAMSTAR) artificial neural networks to predict apnea and hypopnea. Wavelet transform-based preprocessing was applied to six physiological signals obtained from a set of polysomnography studies and used to train and test the networks. MEASUREMENTS AND MAIN RESULTS: We tested prediction performance during non-REM and REM sleep as a function of data segment duration and prediction lead time. Measurements included average sensitivities, specificities, positive predictive values, and negative predictive values. Prediction performed best during non-REM sleep, using 30-second segments to predict events up to 30 seconds into the future. Most events were correctly predicted up to 60 seconds in the future. Apnea prediction achieved a sensitivity and specificity up to 80.6 +/- 5.6 and 72.8 +/- 6.6%, respectively. Hypopnea prediction achieved a sensitivity and specificity up to 74.4 +/- 5.9 and 68.8 +/- 7.0%., respectively. CONCLUSIONS: We report, to our knowledge, the first system to predict individual episodes of apnea and hypopnea. The most important signal for apnea prediction was submental electromyography. The most important signals for hypopnea prediction were submental electromyography and heart rate variability. This prediction system may facilitate improved therapies for obstructive sleep apnea. PMID- 20019340 TI - Impact of untreated obstructive sleep apnea on glucose control in type 2 diabetes. AB - RATIONALE: Obstructive sleep apnea (OSA), a treatable sleep disorder that is associated with alterations in glucose metabolism in individuals without diabetes, is a highly prevalent comorbidity of type 2 diabetes. However, it is not known whether the severity of OSA is a predictor of glycemic control in patients with diabetes. OBJECTIVES: To determine the impact of OSA on hemoglobin A1c (HbA1c), the major clinical indicator of glycemic control, in patients with type 2 diabetes. METHODS: We performed polysomnography studies and measured HbA1c in 60 consecutive patients with diabetes recruited from outpatient clinics between February 2007 and August 2009. MEASUREMENTS AND MAIN RESULTS: A total of 77% of patients with diabetes had OSA (apnea-hypopnea index [AHI] > or =5). Increasing OSA severity was associated with poorer glucose control, after controlling for age, sex, race, body mass index, number of diabetes medications, level of exercise, years of diabetes and total sleep time. Compared with patients without OSA, the adjusted mean HbA1c was increased by 1.49% (P = 0.0028) in patients with mild OSA, 1.93% (P = 0.0033) in patients with moderate OSA, and 3.69% (P < 0.0001) in patients with severe OSA (P < 0.0001 for linear trend). Measures of OSA severity, including total AHI (P = 0.004), rapid eye movement AHI (P = 0.005), and the oxygen desaturation index during total and rapid eye movement sleep (P = 0.005 and P = 0.008, respectively) were positively correlated with increasing HbA1c levels. CONCLUSIONS: In patients with type 2 diabetes, increasing severity of OSA is associated with poorer glucose control, independent of adiposity and other confounders, with effect sizes comparable to those of widely used hypoglycemic drugs. PMID- 20019341 TI - The ratio of FEV1 to FVC as a basis for establishing chronic obstructive pulmonary disease. AB - RATIONALE: The lambda-mu-sigma (LMS) method is a novel approach that defines the lower limit of normal (LLN) for the ratio of FEV1/FVC as the fifth percentile of the distribution of Z scores. The clinical validity of this threshold as a basis for establishing chronic obstructive pulmonary disease is unknown. OBJECTIVE: To evaluate the association between the LMS method of determining the LLN for the FEV1/FVC, set at successively higher thresholds, and clinically meaningful outcomes. METHODS: Using data from a nationally representative sample of 3,502 white Americans aged 40-80 years, we stratified the FEV1/FVC according to the LMS LLN, with thresholds set at the 5th, 10th, 15th, 20th, and 25th percentiles (i.e., LMS-LLN5, LMS-LLN10, etc.). We then evaluated whether these thresholds were associated with an increased risk of death or prevalence of respiratory symptoms. Spirometry was not specifically completed after a bronchodilator. MEASUREMENTS AND MAIN RESULTS: Relative to an FEV1/FVC greater than or equal to LMS-LLN25 (reference group), the risk of death and the odds of having respiratory symptoms were elevated only in participants who had an FEV1/FVC less than LMS LLN(5), with an adjusted hazard ratio of 1.68 (95% confidence interval, 1.34 2.12) and an adjusted odds ratio of 2.46 (95% confidence interval, 2.01-3.02), respectively, representing 13.8% of the cohort. Results were similar for persons aged 40-64 years and those aged 65-80 years. CONCLUSIONS: In white persons aged 40-80 years, an FEV1/FVC less than LMS-LLN5 identifies persons with an increased risk of death and prevalence of respiratory symptoms. These results support the use of the LMS-LLN5 threshold for establishing chronic obstructive pulmonary disease. PMID- 20019343 TI - The integrin-blocking peptide RGDS inhibits airway smooth muscle remodeling in a guinea pig model of allergic asthma. AB - RATIONALE: Airway remodeling, including increased airway smooth muscle (ASM) mass and contractility, contributes to airway hyperresponsiveness in asthma. The mechanisms driving these changes are, however, incompletely understood. Recently, an important role for extracellular matrix proteins in regulating ASM proliferation and contractility has been found, suggesting that matrix proteins and their integrins actively modulate airway remodeling. OBJECTIVES: To investigate the role of RGD (Arg-Gly-Asp)-binding integrins in airway remodeling in an animal model of allergic asthma. METHODS: Using a guinea pig model of allergic asthma, the effects of topical application of the integrin-blocking peptide RGDS (Arg-Gly-Asp-Ser) and its negative control GRADSP (Gly-Arg-Ala-Asp Ser-Pro) were assessed on markers of ASM remodeling, fibrosis, and inflammation induced by repeated allergen challenge. In addition, effects of these peptides on human ASM proliferation and maturation were investigated in vitro. MEASUREMENTS AND MAIN RESULTS: RGDS attenuated allergen-induced ASM hyperplasia and hypercontractility as well as increased pulmonary expression of smooth muscle myosin heavy chain and the proliferative marker proliferating cell nuclear antigen (PCNA). No effects were observed for GRADSP. The RGDS effects were ASM selective, as allergen-induced eosinophil and neutrophil infiltration as well as fibrosis were unaffected. In cultured human ASM cells, we demonstrated that proliferation induced by collagen I, fibronectin, serum, and platelet-derived growth factor requires signaling via RGD-binding integrins, particularly of the alpha(5)beta(1) subtype. In addition, RGDS inhibited smooth muscle alpha-actin accumulation in serum-deprived ASM cells. CONCLUSIONS: This is the first study indicating that integrins modulate ASM remodeling in an animal model of allergic asthma, which can be inhibited by a small peptide containing the RGD motif. PMID- 20019344 TI - Pulmonary alveolar proteinosis in workers at an indium processing facility. AB - Two cases of pulmonary alveolar proteinosis, including one death, occurred in workers at a facility producing indium-tin oxide (ITO), a compound used in recent years to make flat panel displays. Both workers were exposed to airborne ITO dust and had indium in lung tissue specimens. One worker was tested for autoantibodies to granulocytemacrophage-colonystimulating factor (GM-CSF) and found to have an elevated level. These cases suggest that inhalational exposure to ITO causes pulmonary alveolar proteinosis, which may occur via an autoimmune mechanism. PMID- 20019346 TI - Carbon monoxide pollution promotes cardiac remodeling and ventricular arrhythmia in healthy rats. AB - RATIONALE: Epidemiologic studies associate atmospheric carbon monoxide (CO) pollution with adverse cardiovascular outcomes and increased cardiac mortality risk. However, there is a lack of data regarding cellular mechanisms in healthy individuals. OBJECTIVES: To investigate the chronic effects of environmentally relevant CO levels on cardiac function in a well-standardized healthy animal model. METHODS: Wistar rats were exposed for 4 weeks to filtered air (CO < 1 ppm) or air enriched with CO (30 ppm with five peaks of 100 ppm per 24-h period), consistent with urban pollution. Myocardial function was assessed by echocardiography and analysis of surface ECG and in vitro by measuring the excitation-contraction coupling of single left ventricular cardiomyocytes. MEASUREMENTS AND MAIN RESULTS: Chronic CO pollution promoted left ventricular interstitial and perivascular fibrosis, with no change in cardiomyocyte size, and had weak, yet significant, effects on in vivo cardiac function. However, both contraction and relaxation of single cardiomyocytes were markedly altered. Several changes occurred, including decreased Ca(2+) transient amplitude and Ca(2+) sensitivity of myofilaments and increased diastolic intracellular Ca(2+) subsequent to decreased SERCA-2a expression and impaired Ca(2+) reuptake. CO pollution increased the number of arrhythmic events. Hyperphosphorylation of Ca(2+)-handling and sarcomeric proteins, and reduced responses to beta-adrenergic challenge were obtained, suggestive of moderate CO-induced hyperadrenergic state. CONCLUSIONS: Chronic CO exposure promotes a pathological phenotype of cardiomyocytes in the absence of underlying cardiomyopathy. The less severe phenotype in vivo suggests a role for compensatory mechanisms. Arrhythmia propensity may derive from intracellular Ca(2+) overload. PMID- 20019347 TI - "Cherry picking": writing from thin data. PMID- 20019345 TI - Shared treatment decision making improves adherence and outcomes in poorly controlled asthma. AB - RATIONALE: Poor adherence to asthma controller medications results in poor treatment outcomes. OBJECTIVES: To compare controller medication adherence and clinical outcomes in 612 adults with poorly controlled asthma randomized to one of two different treatment decision-making models or to usual care. METHODS: In shared decision making (SDM), nonphysician clinicians and patients negotiated a treatment regimen that accommodated patient goals and preferences. In clinician decision making, treatment was prescribed without specifically eliciting patient goals/preferences. The otherwise identical intervention protocols both provided asthma education and involved two in-person and three brief phone encounters. MEASUREMENTS AND MAIN RESULTS: Refill adherence was measured using continuous medication acquisition (CMA) indices-the total days' supply acquired per year divided by 365 days. Cumulative controller medication dose was measured in beclomethasone canister equivalents. In follow-up Year 1, compared with usual care, SDM resulted in: significantly better controller adherence (CMA, 0.67 vs. 0.46; P < 0.0001) and long-acting beta-agonist adherence (CMA, 0.51 vs. 0.40; P = 0.0225); higher cumulative controller medication dose (canister equivalent, 10.9 vs. 5.2; P < 0.0001); significantly better clinical outcomes (asthma-related quality of life, health care use, rescue medication use, asthma control, and lung function). In Year 2, compared with usual care, SDM resulted in significantly lower rescue medication use, the sole clinical outcome available for that year. Compared with clinician decision making, SDM resulted in: significantly better controller adherence (CMA, 0.67 vs. 0.59; P = 0.03) and long-acting beta-agonist adherence (CMA, 0.51 vs. 0.41; P = 0.0143); higher cumulative controller dose (CMA, 10.9 vs. 9.1; P = 0.005); and quantitatively, but not significantly, better outcomes on all clinical measures. CONCLUSIONS: Negotiating patients' treatment decisions significantly improves adherence to asthma pharmacotherapy and clinical outcomes. Clinical trials registered with www.clinicaltrials.gov (NCT00217945 and NCT00149526). PMID- 20019348 TI - Meeting the complex needs of the health care team: identification of nurse-team communication practices perceived to enhance patient outcomes. AB - Nurses occupy a central position in today's increasingly collaborative health care teams that place a premium on quality patient care. In this study we examined critical team processes and identified specific nurse-team communication practices that were perceived by team members to enhance patient outcomes. Fifty patient-care team members were interviewed to uncover forms of nurse communication perceived to improve team performance. Using a grounded theory approach and constant comparative analysis, study findings reveal two critical processes nurses contribute to as the most central and consistent members of the health care team: ensuring quality decisions and promoting a synergistic team. Moreover, the findings reveal 15 specific nurse-team communication practices that comprise these processes, and thereby are theorized to improve patient outcomes. PMID- 20019349 TI - Sustaining self: the lived experience of transition to long-term ventilation. AB - Long-term mechanical ventilation (LTMV) is used to support growing numbers of children and adults who develop chronic respiratory failure. The purpose of this hermeneutic phenomenological study was to explore the subjective meaning of the experience of transition from spontaneous breathing to reliance on LTMV. In depth, audiorecorded interviews of 11 ventilated individuals living in a western Canadian province were transcribed and analyzed. The transition journey was revealed to be a time of psychological, physical, and spiritual challenge. "Sustaining self " was revealed as the essence of ventilator users' transition experience. Findings underscore the complex nature of transition to technological reliance, illuminating the need for increased health professional knowledge and understanding of the symptom experience and management options for individuals living with chronic respiratory failure. Strategies to support individuals' sense of self as they journey to a life reliant on LTMV are required. PMID- 20019350 TI - Interdisciplinary geriatric and palliative care team narratives: collaboration practices and barriers. AB - Despite the development and implementation of team training models in geriatrics and palliative care, little attention has been paid to the nature and process of teamwork. Geriatrics and palliative care in the clinical setting offer an interdisciplinary approach structured to meet the comprehensive needs of a patient and his or her family. Fellowship members of an interdisciplinary geriatric and palliative care team participated in semistructured interviews. Team members represented social work, chaplaincy, psychology, nursing, and medicine. A functional narrative analysis revealed four themes: voice of the lifeworld, caregiver teamwork, alone on a team, and storying disciplinary communication. The content-ordering function of narratives revealed a divergence in team members' conceptualization of teamwork and team effectiveness, and group ordering of narratives documented the collaborative nature of teams. The study findings demonstrate the potential for narratives as a pedagogical tool in team training, highlighting the benefits of reflective practice for improving teamwork and sustainability. PMID- 20019351 TI - Coping processes as revealed in the stories of mothers of children with autism. AB - Copious research evidence identifies the many stressors faced by mothers of children with autism. The aim of this study was to examine the ways in which coping is revealed in the content and structure of stories told by five mothers of children with autism. Narrative data were analyzed using both holistic-form and categorical-content approaches. Manifestations of coping were revealed in the macrostructures of stories. Cognitive coping strategies were particularly apparent in the life stories, which tended to focus on the emotional and cognitive journeys of the storytellers. Stories of discrete coping episodes added information about behavioral coping strategies employed in specific situations. Analysis of form, particularly of the structure of the life stories, yielded strategies the mothers employed to make meaning of autism in their lives. PMID- 20019352 TI - Chemical carcinogenesis of the gastrointestinal tract in rodents: an overview with emphasis on NTP carcinogenesis bioassays. AB - Cancers of the stomach and large intestine (LI) are the second and fourth leading causes of human cancer mortality. A review of the National Toxicology Program (NTP) database and the Carcinogenic Potency Database (CPDB) reveals that chemically induced neoplasms of the gastrointestinal tract (GIT) are relatively common. Within the GIT, epithelial tumors of the forestomach in mice and rats and LI of the rat are most common. Generally, there is a high species concordance for forestomach with at least 26 chemicals inducing tumors in both species. Glandular stomach tumors are rare, and the few reported are usually neuroendocrine tumors (carcinoids) originating from the enterochromaffin-like (ECL) cells. Of 290 carcinogenic agents identified by the NTP, 19 (7%) caused intestinal neoplasia, 14 in the rat and 5 in the mouse. Neoplasms occurred in both males and females, exclusively in the small intestine (SI) of the mouse and in the LI or both SI and LI in the rat. Enteric carcinogens (NTP) frequently induced neoplasms at other alimentary sites (oral cavity, esophagus, and stomach). In conclusion, the most common induced GIT tumors are squamous neoplasms of the forestomach, glandular neoplasms of the stomach are rare, and rats appear more prone to developing LI (colorectal) cancer compared to mice. PMID- 20019353 TI - Commentary: update on animal models for NTP studies. AB - Based on recommendations of participants at the National Toxicology Program (NTP) workshop "Animal Models for the NTP Cancer Bioassay: Strains and Stocks-Should We Switch?" some modifications to the NTP rodent cancer bioassay were made. The B6C3F1 remains the mouse model used in the cancer bioassay. The use of multiple strains of mice will be explored through a new branch within the NTP, the Host Susceptibility Branch. Several rat models were evaluated; the Harlan Sprague Dawley rat is currently being used in the NTP studies. PMID- 20019354 TI - Precancer: sequentially acquired or predetermined? AB - Recognition of focal morphological intraepithelial lesions associated with the eventual development of invasive cancer has long been the sine qua non of precancer. Empirically, precancers are associated with a morphological continuum from atypia to dysplasia and invasive neoplasia. Such lesions are used as early indicators of cancers and have dramatically reduced mortality from cancers of the colon, uterine cervix, and breast. Progression has been modeled as a linear, stepwise process. Some molecular evidence supports a linear model. However, clinical studies now suggest that preexisting cofactors such as human papilloma virus (HPV) in cervical cancer determines the cell fate. Other clinical studies such as bladder, prostate, and breast suggest that many intraepithelial lesions do not progress to malignancy. The more recent experimental analyses reveal that the key molecular and genetic events even predate the emergence of visible lesions. Thus, a new nonlinear, parallel model is proposed. The parallel model suggests an origin in a putative progenitor cell that expands and invades. The clinical outcome is thus predetermined. If correct, this model suggests that "progression" to malignancy is epigenetic. Further, future assessment of biological potential will involve identification and genetic analysis of the progenitor cell populations. PMID- 20019355 TI - Roles for inflammation and regulatory T cells in colon cancer. AB - Risk for developing cancer rises substantially as a result of poorly regulated inflammatory responses to pathogenic bacterial infections. Anti-inflammatory CD4(+) regulatory cells (T(REG)) function to restore immune homeostasis during chronic inflammatory disorders. It seems logical that T(REG) cells would function to reduce risk of inflammation-associated cancer in the bowel by down-regulating inflammation. It is widely believed, however, that T(REG) function in cancer mainly to suppress protective anticancer inflammatory responses. Thus roles for inflammation, T(REG) cells, and gut bacteria in cancer are paradoxical and are the subject of controversy. Our accumulated data build upon the "hygiene hypothesis" model in which gastrointestinal (GI) infections lead to changes in T(REG) that reduce inflammation-associated diseases. Ability of T(REG) to inhibit or suppress cancer depends upon gut bacteria and IL-10, which serve to maintain immune balance and a protective anti-inflammatory T(REG) phenotype. However, under poorly regulated pro-inflammatory conditions, T(REG) fail to inhibit and may instead contribute to a T helper (Th)-17-driven procarcinogenic process, a cancer state that is reversible by down-regulation of inflammation and interleukin (IL)-6. Consequently, hygienic individuals with a weakened IL-10- and T(REG)-mediated inhibitory loop are highly susceptible to the carcinogenic consequences of elevated inflammation and show more frequent inflammation associated cancers. Taken together, these data help explain the paradox of inflammation and T(REG) in cancer and indicate that targeted stimulation of T(REG) may promote health and significantly reduce risk of cancer. PMID- 20019356 TI - Oxidative stress and oxidative damage in carcinogenesis. AB - Carcinogenesis is a multistep process involving mutation and the subsequent selective clonal expansion of the mutated cell. Chemical and physical agents including those that induce reative oxygen species can induce and/or modulate this multistep process. Several modes of action by which carcinogens induce cancer have been identified, including through production of reactive oxygen species (ROS). Oxidative damage to cellular macromolecules can arise through overproduction of ROS and faulty antioxidant and/or DNA repair mechanisms. In addition, ROS can stimulate signal transduction pathways and lead to activation of key transcription factors such as Nrf2 and NF-kappaB. The resultant altered gene expression patterns evoked by ROS contribute to the carcinogenesis process. Recent evidence demonstrates an association between a number of single nucleotide polymorphisms (SNPs) in oxidative DNA repair genes and antioxidant genes with human cancer susceptibility. These aspects of ROS biology will be discussed in the context of their relationship to carcinogenesis. PMID- 20019357 TI - Visual Arrestin 1 contributes to cone photoreceptor survival and light adaptation. AB - PURPOSE: To evaluate morphologic and functional contributions of Arrestin 1 (Arr1) and Arrestin 4 (Arr4) in cone photoreceptors, the authors examined the phenotypes of visual arrestin knockout mice (Arr1(-/-), Arr4(-/-), Arr1(-/-)Arr4( /-) [Arr-DKO]) reared in darkness. METHODS: Retinal rods and cones were evaluated in wild-type (WT), Arr1(-/-), Arr4(-/-), and Arr-DKO mice using quantitative morphologic analysis, immunoblot, immunohistochemistry, TUNEL, and electroretinographic (ERG) techniques. RESULTS: Compared with either Arr4(-/-) or WT, Arr1(-/-) and Arr-DKO mice had increased apoptotic nuclei in their retinal outer nuclear layer (ONL) at postnatal day (P) 22. By P60, cone density was significantly diminished, but the ONL appeared normal. After 1 minute of background illumination, cone ERG b-wave amplitudes were similar in WT and all Arr KO mice. However, by 3 minutes and continuing through 15 minutes of light adaptation, the cone b-wave amplitudes of WT and Arr4(-/-) mice increased significantly over those of the Arr1(-/-) and Arr-DKO mice, which demonstrated no cone b-wave amplitude increase. In contrast, ERG flicker analysis after the 15 minute light adaptation period demonstrated no loss in amplitude for either Arr1( /-) or Arr4(-/-) mice, whereas Arr-DKO had significantly lower amplitudes. When Arr1 expression was restored in Arr1(-/-) mice (+p48(Arr1-/-)), normal cone density and light-adapted ERG b-wave amplitudes were observed. CONCLUSIONS: In the adult dark-reared Arr1(-/-) and Arr-DKO mice, viable cones diminish over time. Arr1 expression is essential for cone photoreceptor survival and light adaptation, whereas either Arr1 or Arr4 is necessary for maintaining normal flicker responses. PMID- 20019358 TI - Evaluation of experimental autoimmune uveitis in mice treated with FTY720. AB - PURPOSE: FTY720 (fingolimod) is an immunomodulatory drug capable of preventing T cell migration to inflammatory sites by binding to and subsequently downregulating the expression of sphingosine-1 phosphate receptor 1 (S1P(1)) leading in turn to T-cell retention in lymphoid organs. Additional effects of FTY720 by increasing functional activity of regulatory T cells have recently been demonstrated, raising the conversion of conventional T cells into regulatory T cells and affecting the sequestration of regulatory T cells in normal mice. In this study, the action of FTY720 in the ocular autoimmune model in mice was investigated. METHODS: Mice were immunized with 161-180 peptide and pertussis toxin and were treated with 1 mg/kg/d FTY720 by gavage (7-21 days postimmunization [dpi]) or left untreated. Spleen cells, harvested 21 dpi, were cultured and assayed for cytokine production. Draining lymph node, spleen, and eye cells 21 dpi were assayed for quantification of T-cell populations. Disease severity was evaluated by histologic examination of the enucleated eyes at 21 and 49 dpi. In addition, anti-IRBP antibodies were analyzed by ELISA. RESULTS: FTY720 was effective in suppressing the experimental autoimmune uveitis score. Although there was a reduction in the number of eye-infiltrating cells, FTY did not prevent Treg accumulation at this site. FTY720 leads to a significant increase of CD4(+)IFN-gamma(+) and CD4(+)Foxp3(+) cell percentages in lymph nodes, suggesting that this site could be the source of Treg cells found in the eye. CONCLUSIONS: The data showed that treatment in vivo with FTY720 was able to suppress EAU in mice. These results are indicative of the possible therapeutic use of FTY720 in ocular autoimmune processes. PMID- 20019359 TI - Iris pigment epithelial cells express a functional lipopolysaccharide receptor complex. AB - PURPOSE: Ocular pigment epithelial cells are hypothesized to play a role in the pathogenesis of acute anterior uveitis (AAU), where LPS activation of Toll-like receptors (TLRs) may serve as a trigger. In this study, the expression of LPS receptors in iris pigment epithelium (IPE) was determined. METHODS: RT-PCR, flow cytometry, Western blot, and immunohistochemistry were used to investigate the expression of the LPS receptor complex (TLR4, MD-2, and CD14) in primary human IPE. Cytokine secretion by LPS-treated IPE was measured by multiplex bead array and ELISA. The role of CD14 in modulating the LPS response was investigated by addition of soluble CD14 and by antibody neutralization studies. In vivo expression of CD14 was examined by immunohistochemistry and Western blot analysis. RESULTS: IPE expressed TLR4, MD-2, and CD14 in vitro and secreted a panel of proinflammatory cytokines (IL-6, CXCL8, CXCL10, CCL2, CCL4, and CCL5) when stimulated with LPS. CXCL8 secretion by LPS-treated IPE was dependent on CD14 and TLR4. CD14 was detected in CD68+ cells in the iris by immunohistochemistry and in normal aqueous by Western blot analysis. CONCLUSIONS: IPE cells express a functional LPS receptor complex and are capable of promoting ocular inflammation through secretion of an array of proinflammatory mediators. CD14 was identified as a key molecule that modulated the LPS response in IPE. PMID- 20019360 TI - Chemotactic and cytotoxic effects of minocycline on human retinal pigment epithelial cells. AB - PURPOSE: To reveal the effects of minocycline, an anti-inflammatory and neuroprotective agent, on the viability and physiological properties of retinal pigment epithelial (RPE) cells and to compare the effects with those of triamcinolone acetonide. METHODS: The proliferation of human RPE cells in vitro was investigated with a bromodeoxyuridine immunoassay; chemotaxis was examined with a Boyden chamber assay. Cell viability was determined by trypan blue exclusion. The gene expression of growth factors and MMP-9 was determined with real-time RT-PCR, and the secretion of VEGF was examined with ELISA. The phosphorylation of p38 MAPK and ERK1/2 proteins was determined with Western blot analysis. RESULTS: Minocycline at low concentrations (50 nM-20 microM) stimulated chemotaxis and decreased the proliferation of RPE cells. Minocycline at high concentrations (above 5 microM) decreased the viability of RPE cells through the induction of cell necrosis. The chemotactic effect of minocycline was mediated by the stimulation of autocrine PDGF signaling and the activation of p38 MAPK. Minocycline promoted the expression of PDGF-B, HGF, VEGF, and MMP-9 and increased the amounts of phosphorylated p38 and ERK1/2 proteins in RPE cells. Triamcinolone reduced PDGF-evoked chemotaxis and VEGF expression and secretion and had no significant effects on cell viability and proliferation. Triamcinolone did not reverse the effects of minocycline on cell proliferation, chemotaxis, or viability or the expression of VEGF. CONCLUSIONS: Low-dose minocycline induces the activation of RPE cells, as indicated by the activation of p38 and ERK1/2 and by enhanced chemotaxis mediated by autocrine PDGF signaling. High-dose minocycline induces RPE cell degeneration. PMID- 20019361 TI - The role of oxidative stress and inflammation in conjunctivochalasis. AB - Purpose. To investigate the status of oxidative stress and histopathologic alterations in patients with conjunctivochalasis and compare the findings with those in healthy control subjects. Methods. Eleven patients (n = 20 eyes) with Yokoi grade 3 conjunctivochalasis and 11 health control subjects (n = 22 eyes) were prospectively recruited. ELISA for tear hexanoyl-lysine (HEL) and inflammatory cytokines, tear film break-up time tests, Schirmer test measurements, and fluorescein and rose bengal vital staining were performed. Conjunctival specimens obtained during surgery for conjunctivochalasis and cataract underwent immunohistochemical staining for HEL+8-OHdG, MMP-3, and MMP-9, and positively stained cells were counted. Transmission electron microscopy was also performed, with staining for elastic fibers in the conjunctival stroma. Results. The mean tear stability and vital staining scores were significantly worse in the conjunctivochalasis patients than in the control subjects. The tear HEL and tear cytokine levels showed significantly higher values in eyes with conjunctivochalasis. IL-1beta and IL-6 levels showed a significant correlation with corneal epithelial damage. IL-1beta and TNFalpha showed a significant correlation with 8-OHdG-stained cell counts. Specimens from patients with conjunctivochalasis revealed a significantly higher number of cells positively stained for HEL, 8-OHdG, MMP-3, and MMP-9 than did specimens from age- and sex matched control subjects. Transmission electron microscopy showed decreased intercellular cohesiveness, with the conjunctival stroma showing an accumulation of elastic fibers. Conclusions. Lipid and DNA oxidative stress were present in the conjunctiva. Increased tear inflammation seemed to coexist with loss of conjunctival epithelial cohesiveness and increased collagenolytic activity, which may explain the conjunctival laxity observed in patients with conjunctivochalasis. PMID- 20019362 TI - Minimum inhibitory concentrations of standard and novel antimicrobials for isolates from bacterial keratitis. AB - PURPOSE: To determine the minimum inhibitory concentrations (MICs) of 12 antimicrobials in current ophthalmic use and 4 potentially new alternatives against isolates from bacterial keratitis. METHODS: Bacteria were collected from cases of bacterial keratitis in six centers in the United Kingdom between 2003 and 2006. MICs were measured by using susceptibility strips containing a concentration gradient of the antimicrobials penicillin, cefuroxime, ceftazidime, chloramphenicol, gentamicin, amikacin, vancomycin, teicoplanin, ciprofloxacin, ofloxacin, levofloxacin, moxifloxacin, meropenem, linezolid, tigecycline, and daptomycin. RESULTS: Isolates (n = 772) were collected including coagulase negative Staphylococcus (CNS) (30%), Pseudomonas aeruginosa (23%), Staphylococcus aureus (14%), Enterobacteriaceae (14%), and streptococci (13%). Meropenem had low MICs for most isolates. All isolates except P. aeruginosa were susceptible to tigecycline. Linezolid was active against the majority of Gram-positive pathogens. Ten percent of S. aureus and 20% of CNS isolates were methicillin resistant. When systemic breakpoints were used, 84% of S. aureus isolates were susceptible to ciprofloxacin and 98% to moxifloxacin. Of the P. aeruginosa isolates, 99% were susceptible to ceftazidime, 96% to gentamicin, 99% to ciprofloxacin and 100% to moxifloxacin. More than 97% of Enterobacteriaceae isolates were susceptible to ceftazidime, gentamicin, ciprofloxacin, and moxifloxacin. CONCLUSIONS: Based on systemic breakpoint data, resistance to commonly used antimicrobials was apparent. Meropenem is a potentially effective agent for ophthalmic use, with low MICs throughout all the bacterial subgroups. Tigecycline and linezolid showed good activity against particular groups and may be useful for treating bacterial keratitis resistant to current antimicrobials. Of the fluoroquinolones, moxifloxacin showed the lowest MICs and resistance for both Gram-positive and -negative bacteria. PMID- 20019363 TI - Induction of IL-6 in transcriptional networks in corneal epithelial cells after herpes simplex virus type 1 infection. AB - PURPOSE: To determine the transcriptional responses of human corneal epithelial cells (HCECs) after herpes simplex virus type (HSV)-1 infection and to identify the critical inflammatory element(s). METHOD: Immortalized HCECs were infected with HSV-1, and the global transcriptional profile determined. Molecular signaling networks were constructed from the HSV-1-induced transcriptomes. The relationships of the identified networks were confirmed by real-time-PCR and ELISA. Contributions of the critical network nodes were further evaluated by protein array analyses as candidates for inflammatory element induction. RESULTS: HSV-1 infection induced a global transcriptional response, with 412 genes significantly activated or suppressed compared with mock-infected HCECs (P < 0.05, 2< or 0.5> threshold). Infection by UV-inactivated HSV-1 did not induce significant transcriptional activity. Network analysis showed that the HSV-1 induced transcriptomes were associated with JUN N-terminal kinase, p38, extracellular signal-regulated kinase, and nuclear factor kappa-B signaling pathways. These findings indicate that interleukin (IL)-6 and vascular endothelial growth factor (VEGF) probably serve as critical nodes of signaling events. ELISA and protein array analyses verified the induction of the inflammatory elements by HSV infection. Blocking the induction of IL-6 significantly reduced the expression of 21 cytokines, including CCL7, CCL8, CXCL6, transforming growth factor-beta2, platelet-derived growth factor, interferon-gamma, IL-2, and VEGF, thus confirming the critical role of IL-6. CONCLUSIONS: HCECs respond to HSV-1 infection by initiating mitogen-activated protein kinase-related transcriptional events, and IL-6 may serve to induce expression of an array of inflammatory mediators. PMID- 20019364 TI - Vitreous VEGF clearance is increased after vitrectomy. AB - Purpose. Pars plana vitrectomy (PPV) has been reported to reduce macular thickness and improve visual acuity in patients with diabetic macular edema (ME). The hypothesis for the study was that after PPV, clearance is accelerated and VEGF concentrations are reduced. To test this hypothesis, hVEGF(165) injections were performed in rabbit eyes, with and without PPV, and vitreous VEGF levels were measured as a function of time. Methods. The PPV group rabbits had a bilateral 25-gauge PPV, and in the no-PPV group, rabbits had intact vitreous. Intravitreal injections of hVEGF(165) were performed, and the animals were euthanatized at time points up to 7 days. The vitreous was isolated and an enzyme linked immunosorbent assay was used to measure the VEGF levels. Pharmacokinetic parameters were determined in a noncompartmental analysis approach. Results. Mean vitreous VEGF levels decreased more rapidly in eyes subjected to PPV than in no PPV eyes. The vitreous VEGF half-life (t([)(1/2)(])) in PPV eyes was 10 times shorter than that in normal eyes. In addition, mean clearance and mean area under the curve (AUC) increased and decreased, respectively, in eyes that underwent PPV. Conclusions. VEGF clearance is increased after PPV. Reducing VEGF concentrations in the vitreous post-PPV may partially explain the improvement in macular thickness in some patients with ME. Unexpectedly, the half-life of VEGF in the vitreous, even in no-PPV eyes, was <3 hours, whereas compounds of similar molecular weight typically have longer vitreous half-lives. The back of the eye may be uniquely adapted with rapid-clearance mechanisms to regulate vitreous VEGF levels. Further study is suggested. PMID- 20019365 TI - The prostaglandin transporter OATP2A1 is expressed in human ocular tissues and transports the antiglaucoma prostanoid latanoprost. AB - PURPOSE: Latanoprost, a prostaglandin F(2alpha) analogue, has become one of the most widely used medications for the treatment of glaucoma. The authors hypothesized that organic anion transporting polypeptides (OATPs) are responsible for the uptake of latanoprost into ocular tissues and, hence, that they contribute to the interindividual differences in drug concentrations and effects. METHODS: Expression of prostaglandin (PG) transporters (OATP2A1, OATP2B1) in human ocular tissues was determined using real-time RT-PCR and immunofluorescence. The inhibitory interactions between latanoprost and its active metabolite (the free acid) and the uptake of prototypical substrates (PGE(2) and bromosulfophthalein) were tested in stably transfected human embryonic kidney cells overexpressing either OATP2A1 or OATP2B1. These cells were also used to investigate whether latanoprost and latanoprost acid are substrates of OATP2A1 or OATP2B1. RESULTS: OATP2A1 and OATP2B1 mRNA expression was highest in the choroid/retinal pigment epithelium (RPE) complex and ciliary body. OATP2A1 protein expression was most prominent in the RPE and in epithelial and endothelial cell layers of anterior segment tissues, such as cornea, conjunctiva, iris, and ciliary body, whereas OATP2B1 protein was additionally expressed in trabecular meshwork, Schlemm canal, and choroidal vasculature. Latanoprost and latanoprost acid significantly inhibited both OATP2A1 and OATP2B1. Uptake experiments demonstrated that latanoprost acid is effectively transported by OATP2A1 (affinity constant [K(m)], 5.4 microM; maximum uptake rate [V(max)], 21.5 pmol/mg protein/min) and less effectively by OATP2B1. CONCLUSIONS: The results presented herein suggest that at least OATP2A1 plays a role in the intraocular disposition of the therapeutically used prostanoid latanoprost. PMID- 20019366 TI - Reversal of second-hand cigarette smoke-induced impairment of corneal wound healing by thymosin beta4 combined with anti-inflammatory agents. AB - PURPOSE: Abnormalities in corneal reepithelialization caused by second-hand cigarette smoke (CS) are less known than the effects of CS on other tissues. The effects of CS on corneal epithelial cell migration and associated signaling mechanisms were examined, to determine the mechanisms by which CS delays corneal wound healing. METHODS: Corneal epithelial cells in two-dimensional or organ culture were exposed to sidestream whole (SSW) smoke, a major component of second hand CS. Thymosin beta 4 (Tbeta4), a molecule thought to promote wound healing in the cornea, was tested to determine whether it can reverse the adverse effects of SSW smoke on corneal healing. RESULTS: Cell migration, actin reorganization, and phosphorylation of focal adhesion kinase (FAK) and paxillin were all inhibited by exposure to SSW smoke, and the distribution of phospho-src in the cells was disrupted. Activation of RhoA, an important regulator of the cytoskeleton during cell migration, was also inhibited. Tbeta4 stimulated corneal epithelial cell migration in the presence of SSW smoke in culture and in vivo, and it partially reversed the inhibition of corneal healing by SSW smoke. However, Tbeta4 plus dexamethasone, an inhibitor of inflammation, together, reversed the effects of SSW smoke on corneal healing. CONCLUSIONS: These findings suggest that SSW smoke exerts its effects on cell migration during corneal epithelial healing through inhibition of actin reorganization, activation of focal adhesion molecules, formation of the focal adhesion complex, and activation of Rho-GTPases. Furthermore, they strongly suggest that corneal injury induced by toxicants can be treated using anti-inflammatory agents coupled with Tbeta4. PMID- 20019367 TI - Visual noise selectively degrades vision in migraine. AB - Purpose. Migraine is a disabling condition with underlying neuronal mechanisms that remain elusive. Migraineurs experience hyperresponsivity to visual stimuli and frequently experience visual disturbances. In the present study, the equivalent input noise approach was used to reveal abnormalities of visual processing and to isolate factors responsible for any such deficits. This approach partitions visual sensitivity into components that represent the efficiency of using the available stimulus information, the background internal noise due to irregular neuronal fluctuations, and the neuronal noise induced by the external stimulation. Methods. Ten migraine with aura, ten migraine without aura, and ten age-matched headache-free subjects participated. Performance in detecting luminance targets embedded in visual noise, resembling grainy photographs, was measured at various noise levels. Results. Contrast thresholds of the three subject groups were similar in the absence of noise, but both migraine groups performed worse in the presence of high noise levels, with performance of migraineurs with aura significantly poorer (P < 0.05) than that of control subjects. Data were fitted with a perceptual template model that showed that the model parameter determining the internal (neuronal) noise triggered by the external (stimulus) noise was significantly higher (P < 0.001) in both migraine groups than in the non-migraineur group. Migraineurs without aura also showed a significant (P < 0.05) though weak reduction of sampling efficiency (0.12 +/- 0.02) compared with control subjects (0.17 +/- 0.02). Conclusions. The results revealed substantial external noise-exclusion deficits in migraine with aura and a minor impairment of noise exclusion in migraine without aura. Migraineurs appeared prone to abnormally high variability of neuronal activity. This result provides a promising explanation of observed visual deficits in migraine. PMID- 20019369 TI - Pretreatment with intravitreal triamcinolone before laser for diabetic macular edema: 6-month results of a randomized, placebo-controlled trial. AB - PURPOSE: To determine whether pretreatment with intravitreal triamcinolone acetonide (IVTA) before laser photocoagulation is effective in eyes with diabetic macular edema (DME). METHODS: The study was a prospective, double-masked, placebo controlled, clinical trial of eyes with DME and impaired vision (or=5 letters after 6 months. Secondary outcomes were necessity of further treatment, change in central macular thickness, and incidence of adverse events. RESULTS: Eighty-four eyes of 54 participants were entered into the study, with 6-month data available for 81 (96%) of 84 eyes. Improvement of >or=5 logMAR letters was similar in eyes treated with IVTA before laser as placebo (18/42 [43%] IVTA vs. 16/34 [38%] laser alone; P = 0.807), as were retreatment rates at 6 months (22 [56%] IVTA vs. 21 [53%] laser alone; P = 0.727). Mean central macular thickness decreased by 50 microm (95% confidence interval, 10-96 microm) more in the IVTA treatment group than in the laser-alone group after 6 months (P = 0.016). Glaucoma therapy was necessary more frequently in IVTA-treated eyes than in those treated with laser alone (19/42 [45%] vs. 7/42 [17%]; P = 0.005). CONCLUSIONS: Visual results and the need for further laser treatment at 6 months were no better in the IVTA group than in the laser-alone group, despite a better anatomic outcome reflected by reduction in mean central macular thickness. This study found no evidence of a synergistic effect of IVTA and laser photocoagulation for DME. (ClinicalTrials.gov number, NCT00148265). PMID- 20019368 TI - Role of the Fas pathway in Pseudomonas aeruginosa keratitis. AB - PURPOSE: The role of the Fas pathway was tested in Pseudomonas aeruginosa infected mouse cornea by contrasting the responses of FasL(-/-) and wild-type (WT) mice. METHODS: TUNEL staining, real-time RT-PCR, immunostaining, and ELISA assay were used. RESULTS: Compared with WT (resistant) mice, BALB/c FasL(-/-) exhibited significantly elevated bacterial counts and polymorphonuclear leukocyte numbers at 1 and 3 days postinfection (p.i.) and worse outcomes from disease. Similar bacterial challenges in C57BL/6 FasL(-/-) compared with WT mice also led to worsened disease as evidenced by earlier corneal perforation in the susceptible mouse strain. Intense TUNEL staining of apoptotic cells was seen earlier (1 day vs. 3 days) p.i. in BALB/c WT than in knockout mice, This earlier apoptotic pattern correlated with increased expression of caspases 3, 8, and 9 and BAX and with decreased expression of the antiapoptotic molecule Bcl-2. Furthermore, expression levels of the proinflammatory molecule TNF-alpha and its receptor, MIP-2, inducible nitric oxide synthase (iNOS), and nitrite also were significantly elevated in the infected cornea of BALB/c FasL(-/-) compared with WT mice. In vitro, LPS-stimulated Mphi from BALB/c FasL(-/-) mice expressed significantly less caspase 3 and 9, BAX, and IL-10 and more TNF-alpha, MIP-2, and IL-1beta than did cells from WT mice. CONCLUSIONS: Fas-FasL interaction in the cornea balances the host innate immune response to improve disease outcome by promoting earlier apoptosis and regulating proinflammatory cytokines/chemokines and nitric oxide (nitrite) production. Dysregulation of this interaction contributes to bystander tissue damage, enhancing nutrients for bacterial growth and worsened disease outcome after P. aeruginosa infection. PMID- 20019370 TI - Application of a novel interferometric method to investigate the relation between lipid layer thickness and tear film thinning. AB - PURPOSE: The lipid layer of the tear film forms a barrier to evaporation. Evaporation is a major cause of tear thinning between blinks and tear breakup. The purpose of this study was to investigate the relation between tear film thinning and lipid layer thickness before and after instillation of an emulsion eye drop. METHODS: Fifty non-contact lens wearers were studied. Spectral interferometry was used to measure the thinning rate of the precorneal tear film for up to 19 seconds after a blink. Simultaneously, lipid layer thickness was measured based on an absolute reflectance spectrum. After a 2-minute recovery, the measurement was repeated. A drop of the lipid emulsion was then instilled; 15 minutes later, two interferometry measurements were performed similarly. RESULTS: A histogram of thinning rates was fitted by a bimodal distribution with narrow and broad peaks corresponding to slow and rapid thinning, respectively. The correlation between repeated thinning rate measurements was modest, but repeatability was considerably more significant when analyzed in terms of the slow/rapid dichotomy. Similarly, the correlation between thinning rate and lipid thickness was modest but was more evident when analyzed in terms of the slow/rapid dichotomy. Instillation of an emulsion eye drop significantly increased the thickness of the lipid layer but did not significantly alter the thinning rate. CONCLUSIONS: The proposed slow/rapid dichotomy of thinning rates presumably relates to a good/poor barrier to evaporation of the lipid layer. The imperfect correlation between thinning rate and lipid thickness indicates that other factors, such as the composition and structure of the lipid layer, are important (e.g., sufficient polar lipids may be needed to form good interface between nonpolar lipids and the aqueous layer). PMID- 20019371 TI - Formation and disassembly of adherens and tight junctions in the corneal endothelium: regulation by actomyosin contraction. AB - Purpose. To determine the role of actin cytoskeleton in the disassembly and reformation of adherens junctions (AJs) and tight junctions (TJs) in bovine corneal endothelial monolayers. Methods. Disassembly and reformation of AJs and TJs were induced by extracellular Ca(2+) depletion and subsequent add-back of Ca(2+), respectively. Resultant changes in the transendothelial electrical resistance (TER), an indicator of integrity of TJs, were measured based on electrical cell-substrate impedance. Phosphorylated myosin light chain (ppMLC), a biochemical measure of actomyosin contraction, and activation of its upstream regulatory molecule RhoA-GTP were assessed by Western blot analysis. Results. Extracellular Ca(2+) depletion led to activation of RhoA, increase in ppMLC, decrease in TER, contraction of the perijunctional actomyosin ring (PAMR), and redistribution of zonula occludens-1 (ZO-1) and cadherins. These effects were reversed on Ca(2+) add-back. Pretreatment with Y-27632 and blebbistatin (as inhibitors of actomyosin contraction) reduced the rate of decline in TER, opposed the contraction of the PAMR, and blocked the redistribution of ZO-1 and cadherins. Both drugs reduced the recovery in TER and opposed the normal redistribution of ZO-1 and cadherins on Ca(2+) add-back. Cytochalasin D, which led to dissolution of the PAMR, also reduced the recovery of TER on Ca(2+) add back. Conclusions. The (Ca(2+) depletion)-induced disassembly of AJs accelerates the breakdown of TJs through a concomitant increase in the actomyosin contraction of the PAMR. However, these data on reassembly show that a contractile tone of the PAMR is essential for assembly of the apical junctional complex. PMID- 20019372 TI - Differential distribution of blood and lymphatic vessels in the murine cornea. AB - PURPOSE: Because of its unique characteristics, the cornea has been widely used for blood and lymphatic vessel research. However, whether limbal or corneal vessels are evenly distributed under normal or inflamed conditions has never been studied. The purpose of this study was to investigate this question and to examine whether and how the distribution patterns change during corneal inflammatory lymphangiogenesis (LG) and hemangiogenesis (HG). METHODS: Corneal inflammatory LG and HG were induced in two most commonly used mouse strains, BALB/c and C57BL/6 (6-8 weeks of age), by a standardized two-suture placement model. Oriented flat-mount corneas together with the limbal tissues were used for immunofluorescence microscope studies. Blood and lymphatic vessels under normal and inflamed conditions were analyzed and quantified to compare their distributions. RESULTS: The data demonstrate, for the first time, greater distribution of both blood and lymphatic vessels in the nasal side in normal murine limbal areas. This nasal-dominant pattern was maintained during corneal inflammatory LG, whereas it was lost for HG. CONCLUSIONS: Blood and lymphatic vessels are not evenly distributed in normal limbal areas. Furthermore, corneal LG and HG respond differently to inflammatory stimuli. These new findings will shed some light on corneal physiology and pathogenesis and on the development of experimental models and therapeutic strategies for corneal diseases. PMID- 20019373 TI - Regulation of T-cell chemotaxis by programmed death-ligand 1 (PD-L1) in dry eye associated corneal inflammation. AB - PURPOSE. Given that dry eye disease (DED) is associated with T cell-mediated inflammation of the ocular surface and that PD-L1 is an important negative or inhibitory regulator of immune responses constitutively expressed at high levels by corneal epithelial cells, the authors studied the expression and function of PD-L1 in DED. METHODS. Dry eye was induced in untreated wild-type mice, PD-L1(-/ ) mice, and wild-type mice treated with anti-PD-L1 antibody by exposing these mice to a desiccating environment in the controlled environment chamber modified with subcutaneous administration of scopolamine. Real-time PCR was used to quantify the expression of chemokine gene transcript levels of multiple CC and CXC chemokine ligands and receptors. Epifluorescence microscopy was used to evaluate corneal infiltration of CD3(+) T cells after immunohistochemical staining. RESULTS. The increased expression of specific chemokine ligands and receptors in PD-L1(-/-) corneas of normal mice is associated with significant increases in T-cell homing into these corneas. Similar, and more enhanced, increases in T-cell infiltration were observed in PD-L1(-/-) DED mice or DED mice treated with anti-PD-L1 antibody compared with controls. In addition, the authors found significantly decreased expression of PD-L1 by corneal epithelial cells in DED and significantly increased corneal fluorescein staining score with PD-L1 functional blockade using anti-PD-L1 antibody. CONCLUSIONS. Downregulation of corneal epithelial PD-L1 amplifies dry eye-associated corneal inflammation and epitheliopathy by increasing the expression of chemokine ligands and receptors that promote T-cell homing to the ocular surface. PMID- 20019375 TI - Postmenopausal hormone therapy increases retinal blood flow and protects the retinal nerve fiber layer. AB - PURPOSE: To investigate whether postmenopausal hormone therapy (HT) increases retinal and ONH blood flow (BF) and protects ONH topography and the function of retinal ganglion cells in postmenopausal women (PMW). The effect of estradiol (E(2)) treatment on retinal tissue perfusion was also investigated in ovariectomized rats, an animal model for menopause. METHODS: Sixty-four healthy PMW were recruited, 29 of whom never used HT ( HT) and 35 of whom had used HT (+HT) continuously since the onset of menopause. Blood flow of the inferotemporal retinal artery (ITRA), peripapillary retina, and ONH rim were measured in one eye. The ONH stereometric parameters and the pattern electroretinogram (PERG) were also measured. In ovariectomized rats, the retinal tissue perfusion was assessed using the BF tracer N-isopropyl-p-[(14)C]-iodoamphetamine ([(14)C]-IMP) in rats treated with either E(2) (n = 7) or placebo (n = 5). RESULTS: Compared with the HT group, the +HT group presented significantly greater BF of the ITRA (P = 0.006), greater rim volume for the entire ONH region (P = 0.032), and greater rim volume (P = 0.042), height variation contour (P = 0.011), mean thickness (P = 0.033), and cross-sectional area (P = 0.020) of the retinal nerve fiber layer for the inferotemporal region of the ONH when adjusted for age, ocular perfusion pressure, and age at menarche. In ovariectomized rats, E(2) treatment significantly increased retinal perfusion in a range of 22% to 45%. CONCLUSIONS: These findings indicate that estrogens and HT increase retinal blood flow and protect the retinal nerve fiber layer. PMID- 20019376 TI - The breadth and scope of correctional health programs. PMID- 20019374 TI - Onset and progression of with-the-rule astigmatism in children with infantile nystagmus syndrome. AB - PURPOSE: The purpose of this study was to examine the onset and progression of with-the-rule (WTR) astigmatism during the first 8 years of life in children with idiopathic infantile nystagmus syndrome (INS) or INS associated with albinism and to compare their development with that of normal children. Also explored was whether early WTR astigmatism influences emmetropization in children with INS and whether there is evidence of meridional emmetropization. METHODS: Cycloplegic refractions culled from medical records were converted into power vector components: M (spherical equivalent), J(0) (positive J(0) indicates WTR astigmatism), and J(45) (oblique astigmatism). Two diagnostic groups (idiopathic, n = 106; albinism, n = 95) were evaluated and compared with a reference normal group (n = 495). Four age subgroups were evaluated: age< or =0.5 year, 0.535 weeks of gestation (RR 4.3, 95% CI 1.5-12.0), birthweight <2000 g (RR 3.2, 95% CI 1.0-10.3) and onset of jaundice at >60 h of age (RR 3.3, 95% CI 1.2-9.0). Post-phototherapy discharge and follow-up planning should take into account these risk factors. PMID- 20019394 TI - Intermittent short course therapy for pediatric tuberculosis. AB - We conducted this study to assess the efficacy of intermittent short course therapy in all forms of pediatric tuberculosis using a coordinated approach with Revised National Tuberculosis Control Programme (RNTCP). Sixty five children were treated using RNTCP protocols with some modifications, such as dose adjustments or prolongation of treatment in selected children. Overall response rate was 95%(pulmonary 94% and extra pulmonary 97%). There was one case with possible relapse. With dynamic inputs from both the treating pediatrician and personnel from Directly Observed Treatment Short course (DOTS) centers, we could successfully implement RNTCP protocols in childhood tuberculosis. PMID- 20019395 TI - Clinico-radiological correlation in childhood hypopituitarism. AB - Non-tumor etiology constitutes a major group of childhood hypopituitarism. Magnetic resonance imaging has enormously complimented hormonal assessment in these patients. We describe clinico-radiological correlates in thirty-one children (23 boys), aged 1-17 years with a peak GH (growth hormone) levels <7 ng/mL after pharmacological stimuli. Hypoplastic pituitary gland was the most frequent abnormality in children with isolated growth hormone deficiency (IGHD) as compared to stalk abnormalities in children with multiple pituitary hormone deficiencies. MRI tetrad (hypoplastic/absent pituitary, hypoplastic stalk, absent/ectopic posterior pituitary bright spot and empty sella) was more prevalent in IGHD. MRI abnormalities correlated with the severity of growth hormone deficiency. PMID- 20019396 TI - Effect of backpack weight on postural angles in preadolescent children. AB - BACKGROUND: Carrying heavy backpacks could cause a wide spectrum of pain related musculoskeletal disorders and postural dysfunctions. OBJECTIVE: To determine the changes in various postural angles with different backpack weights in preadolescent children. DESIGN: Cross-sectional. PARTICIPANTS: Healthy male school-children (n=200), mean (SD) age: 12.5 (0.5) years, from high schools in Mangalore, India. MEASUREMENTS: Bodyweight and height were measured using a forceplate and stadiometer, respectively. From the weight recorded, 5%, 10%, 15%, 20%, and 25% of the bodyweight were calculated and implemented as their respective backpack loads. The Image Tool version 3.0, digitizing software was used for analyzing photographs to determine craniovertebral (CV), head on neck (HON), head and neck on trunk (HNOT), trunk and lower limb angles. Postural angles were compared with no backpack and with backpacks weighing 5% to 25% of the subject's bodyweight. RESULTS: The CV angle changed significantly after 15% of backpack load (P <0.05). The HON and HNOT angles changed significantly after 10% of backpack load (P <0.05). The trunk and lower limb angle also changed significantly after 5% of backpack load (P <0.05). CONCLUSIONS: Carrying a backpack weighing 15% of body weight change all the postural angles in preadolescent children. PMID- 20019397 TI - Hypovitaminosis d and hypocalcemic seizures in infancy. AB - BACKGROUND: Hypocalcemia accounts for a majority of seizures in infants reporting to the emergency ward of our hospital. OBJECTIVE: To evaluate the role of Vitamin D deficiency in the etiology of hypocalcemic seizures in infancy. DESIGN AND SETTING: Cross sectional hospital based study, from April 2006-March 2007. SUBJECTS: 60 infants with hypocalcemic seizures and their mothers (study group) and 60 healthy breastfed infants with their lactating mothers (control group). MEASUREMENTS: Vitamin D [25(OH) D] and intact para-thormone levels. RESULTS: High prevalence of hypovitaminosis D [25(OH)D levels <10 ng/mL] was observed in study mothers (85%), control mothers (50%), study infants (90%), and control infants (41.7%). Mean serum 25(OH) D values in study mothers and their infants (6.54 +/- 5.32 ng/mL and 4.92 +/- 4.62 ng/mL) were significantly lower than those of mother infant pairs (9.06 +/- 4.78 ng/mL and 9.03 +/- 4.63 ng/mL) in the control group (P<0.001). A strong positive correlation of 25(OH) D levels between mothers and their infants was seen in both the study and control populations (P<0.001). Of the 54 study infants who had 25(OH)D levels <10 ng/mL, 48 (89%) were born to mothers who also had 25(OH) D levels <10 ng/mL. CONCLUSIONS: Vitamin D deficiency is a major cause of hypocalcemic seizures in infants. Infants born to vitamin D deficient mothers are at a significantly higher risk to develop hypocalcemic seizures. PMID- 20019398 TI - A novel method for patient exit and entrance dose prediction based on water equivalent path length measured with an amorphous silicon electronic portal imaging device. AB - In vivo dosimetry is one of the quality assurance tools used in radiotherapy to monitor the dose delivered to the patient. Electronic portal imaging device (EPID) images for a set of solid water phantoms of varying thicknesses were acquired and the data fitted onto a quadratic equation, which relates the reduction in photon beam intensity to the attenuation coefficient and material thickness at a reference condition. The quadratic model is used to convert the measured grey scale value into water equivalent path length (EPL) at each pixel for any material imaged by the detector. For any other non-reference conditions, scatter, field size and MU variation effects on the image were corrected by relative measurements using an ionization chamber and an EPID. The 2D EPL is linked to the percentage exit dose table, for different thicknesses and field sizes, thereby converting the plane pixel values at each point into a 2D dose map. The off-axis ratio is corrected using envelope and boundary profiles generated from the treatment planning system (TPS). The method requires field size, monitor unit and source-to-surface distance (SSD) as clinical input parameters to predict the exit dose, which is then used to determine the entrance dose. The measured pixel dose maps were compared with calculated doses from TPS for both entrance and exit depth of phantom. The gamma index at 3% dose difference (DD) and 3 mm distance to agreement (DTA) resulted in an average of 97% passing for the square fields of 5, 10, 15 and 20 cm. The exit dose EPID dose distributions predicted by the algorithm were in better agreement with TPS calculated doses than phantom entrance dose distributions. PMID- 20019399 TI - LET dependence of GafChromic films and an ion chamber in low-energy proton dosimetry. AB - Dosimetry using a PMMA phantom was performed in 15 and 29 MeV proton beams from the Birmingham cyclotron, with a Markus parallel-plate ionization chamber and GafChromic EBT and MD-V2-55 film. Simulations of the depth-dose curves were performed with FLUKA 2008.3 and MCNPX 2.5.0, which agreed almost perfectly with each other in range and only differed by 2% in the Bragg peak (BP) region. FLUKA was also used to calculate k(Q) factors for Markus chamber measurements as an improvement to the IAEA TRS-398 values in low-energy beams. FLUKA depth-dose simulations overestimate the BP height measured by ion chamber by about 10%, where the initial proton energy spread was estimated by fitting to the slope of the measured BP distal edge. Both GafChromic films showed an under-response in the BP compared to ion chamber; however, EBT exhibits this effect at lower energies than MD-V2-55. A possible reason for this is attributed to the shape and arrangement of the monomer particles being different in the active components of EBT and MD-V2-55. Relative effectiveness (RE) of both films is presented as functions of residual range R(res) in water and peak proton energy determined by FLUKA, with considerations for the spatial separation of the two active layers in each film. The proton energies at which RE reduces to 90% of maximum film response are 6.7 and 3.2 MeV for MD-V2-55 and EBT, respectively. Additionally, a beam quality correction factor (g(Q,Q0)) is suggested for both GafChromic films, involving water-to-film stopping power ratios evaluated using ICRU recommendations, and a polymer yield factor G(Q0)/G(Q). RE in this work is equated to the reciprocal of the polymer yield factor. The calculated values of (S(w,film))Q/(S(w,film))Q0 are constant within 2.1% and 1.2% across the proton energy range of 1-300 MeV for EBT and MD-V2-55, respectively, so it is concluded that the polymer yield factor is the dominant factor causing the LET quenching effect. PMID- 20019401 TI - The UF family of reference hybrid phantoms for computational radiation dosimetry. AB - Computational human phantoms are computer models used to obtain dose distributions within the human body exposed to internal or external radiation sources. In addition, they are increasingly used to develop detector efficiencies for in vivo whole-body counters. Two classes of computational human phantoms have been widely utilized for dosimetry calculation: stylized and voxel phantoms that describe human anatomy through mathematical surface equations and 3D voxel matrices, respectively. Stylized phantoms are flexible in that changes to organ position and shape are possible given avoidance of region overlap, while voxel phantoms are typically fixed to a given patient anatomy, yet can be proportionally scaled to match individuals of larger or smaller stature, but of equivalent organ anatomy. Voxel phantoms provide much better anatomical realism as compared to stylized phantoms which are intrinsically limited by mathematical surface equations. To address the drawbacks of these phantoms, hybrid phantoms based on non-uniform rational B-spline (NURBS) surfaces have been introduced wherein anthropomorphic flexibility and anatomic realism are both preserved. Researchers at the University of Florida have introduced a series of hybrid phantoms representing the ICRP Publication 89 reference newborn, 15 year, and adult male and female. In this study, six additional phantoms are added to the UF family of hybrid phantoms-those of the reference 1 year, 5 year and 10 year child. Head and torso CT images of patients whose ages were close to the targeted ages were obtained under approved protocols. Major organs and tissues were segmented from these images using an image processing software, 3D-DOCTOR. NURBS and polygon mesh surfaces were then used to model individual organs and tissues after importing the segmented organ models to the 3D NURBS modeling software, Rhinoceros. The phantoms were matched to four reference datasets: (1) standard anthropometric data, (2) reference organ masses from ICRP Publication 89, (3) reference elemental compositions provided in ICRP 89 as well as ICRU Report 46, and (4) reference data on the alimentary tract organs given in ICRP Publications 89 and 100. Various adjustments and refinements to the organ systems of the previously described newborn, 15 year and adult phantoms are also presented. The UF series of hybrid phantoms retain the non-uniform scalability of stylized phantoms while maintaining the anatomical realism of patient-specific voxel phantoms with respect to organ shape, depth and inter-organ distance. While the final versions of these phantoms are in a voxelized format for radiation transport simulation, their primary format is given as NURBS and polygon mesh surfaces, thus permitting one to sculpt non-reference phantoms using the reference phantoms as an anatomic template. PMID- 20019400 TI - MR-guided transcranial brain HIFU in small animal models. AB - Recent studies have demonstrated the feasibility of transcranial high-intensity focused ultrasound (HIFU) therapy in the brain using adaptive focusing techniques. However, the complexity of the procedures imposes provision of accurate targeting, monitoring and control of this emerging therapeutic modality in order to ensure the safety of the treatment and avoid potential damaging effects of ultrasound on healthy tissues. For these purposes, a complete workflow and setup for HIFU treatment under magnetic resonance (MR) guidance is proposed and implemented in rats. For the first time, tissue displacements induced by the acoustic radiation force are detected in vivo in brain tissues and measured quantitatively using motion-sensitive MR sequences. Such a valuable target control prior to treatment assesses the quality of the focusing pattern in situ and enables us to estimate the acoustic intensity at focus. This MR-acoustic radiation force imaging is then correlated with conventional MR-thermometry sequences which are used to follow the temperature changes during the HIFU therapeutic session. Last, pre- and post-treatment magnetic resonance elastography (MRE) datasets are acquired and evaluated as a new potential way to non-invasively control the stiffness changes due to the presence of thermal necrosis. As a proof of concept, MR-guided HIFU is performed in vitro in turkey breast samples and in vivo in transcranial rat brain experiments. The experiments are conducted using a dedicated MR-compatible HIFU setup in a high-field MRI scanner (7 T). Results obtained on rats confirmed that both the MR localization of the US focal point and the pre- and post-HIFU measurement of the tissue stiffness, together with temperature control during HIFU are feasible and valuable techniques for efficient monitoring of HIFU in the brain. Brain elasticity appears to be more sensitive to the presence of oedema than to tissue necrosis. PMID- 20019402 TI - The Virtual Family--development of surface-based anatomical models of two adults and two children for dosimetric simulations. AB - The objective of this study was to develop anatomically correct whole body human models of an adult male (34 years old), an adult female (26 years old) and two children (an 11-year-old girl and a six-year-old boy) for the optimized evaluation of electromagnetic exposure. These four models are referred to as the Virtual Family. They are based on high resolution magnetic resonance (MR) images of healthy volunteers. More than 80 different tissue types were distinguished during the segmentation. To improve the accuracy and the effectiveness of the segmentation, a novel semi-automated tool was used to analyze and segment the data. All tissues and organs were reconstructed as three-dimensional (3D) unstructured triangulated surface objects, yielding high precision images of individual features of the body. This greatly enhances the meshing flexibility and the accuracy with respect to thin tissue layers and small organs in comparison with the traditional voxel-based representation of anatomical models. Conformal computational techniques were also applied. The techniques and tools developed in this study can be used to more effectively develop future models and further improve the accuracy of the models for various applications. For research purposes, the four models are provided for free to the scientific community. PMID- 20019403 TI - Development of a frameless stereotactic radiosurgery system based on real-time 6D position monitoring and adaptive head motion compensation. AB - Stereotactic radiosurgery delivers radiation with great spatial accuracy. To achieve sub-millimeter accuracy for intracranial SRS, a head ring is rigidly fixated to the skull to create a fixed reference. For some patients, the invasiveness of the ring can be highly uncomfortable and not well tolerated. In addition, placing and removing the ring requires special expertise from a neurosurgeon, and patient setup time for SRS can often be long. To reduce the invasiveness, hardware limitations and setup time, we are developing a system for performing accurate head positioning without the use of a head ring. The proposed method uses real-time 6D optical position feedback for turning on and off the treatment beam (gating) and guiding a motor-controlled 3D head motion compensation stage. The setup consists of a central control computer, an optical patient motion tracking system and a 3D motion compensation stage attached to the front of the LINAC couch. A styrofoam head cast was custom-built for patient support and was mounted on the compensation stage. The motion feedback of the markers was processed by the control computer, and the resulting motion of the target was calculated using a rigid body model. If the target deviated beyond a preset position of 0.2 mm, an automatic position correction was performed with stepper motors to adjust the head position via the couch mount motion platform. In the event the target deviated more than 1 mm, a safety relay switch was activated and the treatment beam was turned off. The feasibility of the concept was tested using five healthy volunteers. Head motion data were acquired with and without the use of motion compensation over treatment times of 15 min. On average, test subjects exceeded the 0.5 mm tolerance 86% of the time and the 1.0 mm tolerance 45% of the time without motion correction. With correction, this percentage was reduced to 5% and 2% for the 0.5 mm and 1.0 mm tolerances, respectively. PMID- 20019404 TI - A methodology for selecting the beam arrangement to reduce the intensity modulated radiation therapy (IMRT) dose to the SPECT-defined functioning lung. AB - Macroaggregated albumin single-photon emission computed tomography (MAA-SPECT) provides a map of the spatial distribution of lung perfusion. Our previous work developed a methodology to use SPECT guidance to reduce the dose to the functional lung in IMRT planning. This study aims to investigate the role of beam arrangement on both low and high doses in the functional lung. In our previous work, nine-beam IMRT plans were generated with and without SPECT guidance and compared for five patients. For the current study, the dose-function histogram (DFH) contribution for each of the nine beams for each patient was calculated. Four beams were chosen based on orientation and DFH contributions to create a SPECT-guided plan that spared the functional lung and maintained target coverage. Four-beam SPECT-guided IMRT plans reduced the F(20) and F(30) values by (16.5 +/- 6.8)% and (6.1 +/- 9.2)%, respectively, when compared to nine-beam conventional IMRT plans. Moreover, the SPECT-4F Plan reduces F(5) and F(13) for all patients by (11.0 +/- 8.2)% and (6.1 +/- 3.6)%, respectively, compared to the SPECT Plan. Using fewer beams in IMRT planning may reduce the amount of functional lung that receives 5 and 13 Gy, a factor that has recently been associated with radiation pneumonitis. PMID- 20019405 TI - An agent-based model identifies MRI regions of probable tumor invasion in a patient with glioblastoma. AB - We present an application of a previously developed agent-based glioma model (Chen et al 2009 Biosystems 95 234-42) for predicting spatio-temporal tumor progression using a patient-specific MRI lattice derived from apparent diffusion coefficient (ADC) data. Agents representing collections of migrating glioma cells are initialized based upon voxels at the outer border of the tumor identified on T1-weighted (Gd+) MRI at an initial time point. These simulated migratory cells exhibit a specific biologically inspired spatial search paradigm, representing a weighting of the differential contribution from haptotactic permission and biomechanical resistance on the migration decision process. ADC data from 9 months after the initial tumor resection were used to select the best search paradigm for the simulation, which was initiated using data from 6 months after the initial operation. Using this search paradigm, 100 simulations were performed to derive a probabilistic map of tumor invasion locations. The simulation was able to successfully predict a recurrence in the dorsal/posterior aspect long before it was depicted on T1-weighted MRI, 18 months after the initial operation. PMID- 20019406 TI - Mixing processes in the vitreous chamber induced by eye rotations. AB - In this paper, we study a model of flow in the vitreous humour in the posterior chamber of the human eye, induced by saccadic eye rotations. We concentrate on the effect of the shape of the chamber upon the mixing properties of the induced flows. We make particle image velocimetry measurements of the fluid velocity in a transparent plastic (Perspex) model of the posterior chamber during sinusoidal torsional oscillations about a vertical axis. We use a Newtonian fluid to model the vitreous humour, which is most realistic when either the vitreous humour is liquefied or has been replaced by purely viscous tamponade fluids. The model of the posterior chamber is a sphere with an indentation, representing the effect of the lens. In spite of the purely periodic forcing, a steady streaming flow is generated, which plays a fundamental role in the mixing processes in the domain. The streaming flow differs markedly from that in a perfect sphere, and its topological characteristics change substantially as the frequency of oscillation varies. We discuss the flow characteristics in detail and show that, for physiological parameter values, the Peclet number (based on a suitable measure of the steady streaming velocity) is large, suggesting that advection strongly dominates over diffusion for mass transport phenomena. We also compute particle trajectories based on the streaming velocity and use these to investigate the stirring properties of the flow. PMID- 20019407 TI - The initiation of the heart beat. AB - During a normal lifetime, the heart may beat over 2 billion times, but the mechanisms by which the heart beats are initiated remain a subject of intense investigation. Since the discovery of a pacemaker current (I(f)) in 1978, multiple studies have shown that rhythmic changes in membrane voltage (the "membrane voltage clock") underlie the mechanisms of automaticity. The I(f) is a depolarization current activated during hyperpolarization. Therefore, when the cardiac cells recover, the I(f) is activated and slowly depolarizes the cell membrane, leading to the onset of action potential. Recent studies, however, suggest that increased intracellular Ca (Ca(i)) induced by spontaneous rhythmic sarcoplasmic reticulum Ca release (the "calcium clock") is also jointly responsible for the initiation of the heart beat. Elevated Ca(i) activates another ionic current (the sodium-calcium exchanger current or I(NCX)), leading to spontaneous phase 4 depolarization. Under normal conditions, both clocks are needed to initiate the heart beat. Malfunction of the clocks is associated with sinus node dysfunction in heart failure and atrial fibrillation. More studies are needed to determine how both clocks work together to initiate heart beat under normal and disease conditions. PMID- 20019408 TI - Iron regulatory hormone hepcidin decreases in chronic heart failure patients with anemia. AB - BACKGROUND: The etiology of anemia is still unclear in patients with chronic heart failure (CHF). Hepcidin is an iron regulatory peptide that is synthesized in the liver to suppress iron absorption and utilization. Hepcidin synthesis is suppressed by anemia, hypoxia and erythropoiesis, and induced by inflammation. Inflammatory cytokines, such as interleukin-6 (IL-6), increase the synthesis of hepcidin, resulting in anemia of inflammation (AI). The serum hepcidin concentration in CHF patients with anemia was measured in order to better understand anemia in CHF. METHODS AND RESULTS: Serum hepcidin-25, erythropoietin (EPO), ferritin and IL-6 concentrations were measured in 61 CHF patients. Among these patients, 36 patients had anemia. A group of 16 patients without cardiac disease or anemia were recruited as controls. Serum IL-6 and EPO were higher and hepcidin-25 was lower in CHF patients with anemia than in controls. Hepcidin-25 correlated with EPO and ferritin but not with IL-6. Results of multivariable regression analysis showed that independent predictors of serum hepcidin-25 included EPO and ferritin but not IL-6. CONCLUSIONS: Serum hepcidin-25 concentrations were regulated by iron storage and erythropoiesis but not by IL-6 in CHF patients with anemia. These findings might indicate that AI is a minor cause of anemia in CHF. PMID- 20019410 TI - Prevalence of the Brugada-type electrocardiogram and incidence of Brugada syndrome in patients with sick sinus syndrome. AB - BACKGROUND: In the present study, clarification of the prevalence of the Brugada type electrocardiogram (ECG) and the incidence of spontaneous ventricular fibrillation (VF) that occurred with the Brugada-type ECG in patients with sick sinus syndrome (SSS) was determined. METHODS AND RESULTS: A total of 487 consecutive patients (men 45%, mean age 69.9+/-12.3 years), who were defined as having an indication for cardiac pacemaker (PM) for SSS, were investigated. The ECG before an initial PM implantation and occurrence of VF or sudden cardiac death (SCD) was examined retrospectively. Brugada-type ECG was found in 14 patients (2.87%) including 4 (0.82%) with type 1 and 10 (2.05%) with type 2. During the follow-up period of 7.2+/-5.4 years, 2 out of the 4 patients with type 1 ECG had experienced a VF episode after the device implantation. In 10 patients with type 2 ECG, none had VF or SCD. The incidence of spontaneous VF (Brugada syndrome) in SSS patients was calculated as 14.1 per 100 person-years with type 1 ECG. CONCLUSIONS: The prevalence of typical Brugada-type (type 1) ECG in SSS patients seems to be higher compared with the general population. In addition, SSS patients with the typical Brugada-type ECG might be a high risk for spontaneous VF. PMID- 20019409 TI - Latest management and outcomes of major pulmonary embolism in the cardiovascular disease early transport system: Tokyo CCU Network. AB - BACKGROUND: Major pulmonary embolism (PE) is a life-threatening disorder associated with high mortality and morbidity. The clinical characteristics and outcomes in major PE managed by a well-organized cardiac care regional urban network and hospitals have not been clarified and were examined in the present study. METHODS AND RESULTS: Data from the Tokyo CCU Network registered cohort in 2005-2006 were analyzed. Among 193 patients with major PE and known severities and outcomes, 42 patients had massive PE, defined as cardiogenic shock or cardiac arrest. The median time from symptom onset to CCU admission was 16.3 h. The in hospital mortality of the 124 patients who received reperfusion therapy was lower than that of the 69 patients that did not receive reperfusion therapy (11.3% vs 18.8%; P=0.15). In multiple logistic regression analyses after adjusting for advanced age and sex, reperfusion therapy was selected as a significant predictor for in-hospital death (adjusted odds ratio, 0.34; 95%CI, 0.12-0.95; P=0.039), in addition to massive type (adjusted odds ratio, 14.02; 95%CI, 4.71-41.76; P<0.0001). CONCLUSIONS: Early transport and specific reperfusion therapy for major PE were effectively performed by the Tokyo CCU Network, suggesting the efficacy of a specialty management system for major PE. PMID- 20019411 TI - Association between green tea intake and coronary artery disease in a Chinese population. AB - BACKGROUND: There is still conflicting evidence that green tea may protect against coronary atherosclerosis therefore the present study investigated the association between green tea consumption and arteriographically determined coronary atherosclerosis in a Chinese population. METHODS AND RESULTS: The study population consisted of 520 consecutive patients (379 men and 141 women) who underwent coronary arteriography for the first time. Patients were divided into 2 groups (Non-coronary artery disease [CAD] and CAD groups) according to the results of coronary arteriography. After adjusting the established and potential confounders, green tea consumption was associated with a reduced risk of CAD in male patients, with an adjusted odds ratio (OR) of 0.62 (95% confidence interval, 0.38-1.01) compared with those who did not drink green tea. Compared to non-tea drinkers, the adjusted ORs were 1.09 (0.61-1.96) in male patients consuming less than 125 g of dried green tea leaves per month, 0.36 (0.19-0.71) for 125-249 g per month and 0.36 (0.17-0.73) for > or =250 g per month, with a statistically significant test for trend (P<0.001). Similar dose-response relationships were also observed for frequency, duration, concentration and starting age of green tea drinking in male patients. In female patients, no inverse association was found between green tea consumption and CAD. CONCLUSIONS: Green tea consumption can protect against the development of coronary atherosclerosis in Chinese male patients. PMID- 20019412 TI - Coronary angioscopic evaluation for serial changes of luminal appearance after pharmacological and catheter interventions. AB - Although preventive pharmacological therapies effectually reduce the risk of cardiovascular events, acute coronary syndrome (ACS) remains a leading cause of morbidity and mortality in our country, Japan. Disruption of atherosclerotic vulnerable plaques and flow-limiting thrombus formation in non-stent segments of native coronary arteries are considered a main mechanism of ACS. In addition, stent thrombosis originating from implanted metallic coronary stents, so-called vulnerable stents, occasionally appears as ACS in the clinical settings. Coronary angioscopy is a unique imaging modality permitting direct visualization of luminal structures, such as atherosclerotic plaque, thrombus, stent struts, and proliferating neointima. On the basis of accumulated angioscopic findings, intense yellow plaques and stents without neointimal coverage are considered vulnerable plaques and vulnerable stents, respectively. In contrast, morphological disappearance of vulnerable plaques or vulnerable stents by pharmacological and trans-catheter therapies imply stabilization of the plaques or stents. Hence, angioscopic assessment for vulnerability (or stability) of atherosclerotic plaques and implanted stents might be useful for risk classification in the future events of ACS. To evaluate serial changes of coronary lumen after pharmacological and catheter interventions using angioscopy might also provide important information on potential benefits and surrogate endpoints of the therapies and on patients' management. PMID- 20019413 TI - Documentation of impaired coronary blood flow in chronic obstructive pulmonary disease patients. AB - BACKGROUND: In the current study, the effects of moderate to severe chronic obstructive pulmonary disease (COPD) on coronary blood flow in patients with angiographically proven normal coronary arteries was evaluated. METHODS AND RESULTS: A total of 85 patients with moderate to severe COPD and 39 age- and sex matched control partcipants, who underwent diagnostic coronary angiography and found to have normal epicardial coronary angiogram constituted the COPD and control groups, respectively. The 2 groups were compared for Thrombolysis In Myocardial Infarction (TIMI) frame counts in each major coronary artery. The TIMI frame count of the COPD group was significantly higher than that of control group for all 3 major individual coronary arteries: left anterior descending (corrected), 37+/-13 vs 20+/-4; right coronary artery, 32+/-14 vs 21+/-4; and left circumflex artery, 34+/-12 vs 20+/-5, (P<0.001 for all). In addition, TIMI frame counts in individual coronary arteries were found to be positively correlated with forced expiratory volume 1 s percent, serum high sensitive C reactive protein and fibrinogen concentrations, in the COPD group. CONCLUSIONS: Our findings suggest that an increased slow coronary flow might be a manifestation of harmful effects of COPD on the coronary circulation, regardless of the underlying mechanism. PMID- 20019414 TI - Comparison of neointimal coverage of sirolimus-eluting stents and paclitaxel eluting stents using optical coherence tomography at 9 months after implantation. AB - BACKGROUND: The differences between using sirolimus-eluting stents (SESs) and paclitaxel-eluting stents (PESs) in the vascular response at 9 months after implantation were examined with optical coherence tomography (OCT). METHODS AND RESULTS: OCT was carried out in 33 SESs [33 patients, 19 with acute coronary syndrome (ACS) and 14 with stable angina pectoris (SAP)] and 27 PESs (27 patients, 15 with ACS and 12 with SAP) at 9 months after stent implantation. Stent strut coverage and apposition at each strut were evaluated. The frequency of uncovered struts was significantly higher in SES (12.5+/-15.2 vs 4.9+/-7.9 %, P=0.01). The incidence of complete covered stents with neointima was 9.1% (3/33) in SES and 29.6% (8/27) in PES (P=0.05). The pattern of neointima in PES was more heterogeneous than that in SES (1.3+/-0.5 for SES vs 2.0+/-0.6 for PES, P<0.001). The intracoronary thrombus was frequently detected in SES [10 (30.3%) in SES vs 5 (18.5%) in PES, P=0.29]. CONCLUSIONS: Uncovered struts were frequently observed in SES, but the pattern of neointima was more heterogeneous in PES at 9 months. In addition, stent coverage was incomplete in both stent groups at 9 months after stent implantation. PMID- 20019415 TI - Upregulation of fractalkine and its receptor, CX3CR1, is associated with coronary plaque rupture in patients with unstable angina pectoris. AB - BACKGROUND: Recent studies suggest that fractalkine (FKN or CX3CL1) and its cognate receptor, CX3CR1, play a role in atherogenesis, so the relationship between coronary plaque rupture, as observed by preintervention optical coherence tomography, and plasma levels of FKN and CX3CR1 was investigated in this study. METHODS AND RESULTS: The study population consisted of 46 patients with unstable angina pectoris (UAP), 30 patients with stable angina pectoris, and 25 healthy controls. The UAP patients underwent a preintervention optical coherence tomography study, which revealed that the number of patients with and without plaque rupture at the culprit site was 27 (rupture group) and 19 (non-rupture group), respectively. Plasma levels of soluble FKN (sFKN) and CX3CR1 were measured by enzyme-linked immunosorbent assay and flow cytometry, respectively. The plasma levels of sFKN were significantly increased in UAP patients with plaque rupture compared with patients in the other groups. Multiple logistic regression analysis showed that CD14(+)CD16(+)CX3CR1(+) monocytes and CD3(+)CX3CR1(+) lymphocytes were independent predictors of the presence of ruptured plaque. CONCLUSIONS: Increases in the FKN level and the number of CX3CR1 expressing mononuclear cells might contribute to coronary plaque rupture. PMID- 20019416 TI - Effects of different ceramic and composite materials on stress distribution in inlay and onlay cavities: 3-D finite element analysis. AB - To reduce loss of tooth tissue and to improve esthetic results, inlay and onlay restorations are good treatment choices for extensive cavities in posterior teeth. The aim of this paper was to evaluate, by means of three-dimensional finite element analysis, the effects of restorative material and cavity design on stress distribution in the tooth structures and restorative materials. Two different nanofilled composites and two different all-ceramic materials were used in this study. A permanent molar tooth was modeled with enamel and dentin structures. 3-D inlay and onlay cavity designs were created. Von Mises, compressive, and tensile stresses on the restorative materials, core materials, enamel, and dentin were evaluated separately. On the effect of restorative material, results showed that in the case of materials with low elastic moduli, more stress was transferred to the tooth structures. Therefore, compared to the nanofilled composites, the all-ceramic inlay and onlay materials tested transferred less stress to the tooth structures. On the effect of cavity design, the onlay design was more efficacious in protecting the tooth structures than the inlay design. PMID- 20019417 TI - Finite element analysis of weakened roots restored with composite resin and posts. AB - Finite element analysis (FEA) was used to investigate the influence of different post systems on the stress distribution of weakened teeth under oblique-load application. A maxillary central incisor root obtained from a sound tooth was weakened by partial removal of dentin inside the root canal. Seven two dimensional numerical models, one from the sound tooth and six from the weakened root restored with composite resin and post systems were created as follows - ST: sound tooth; CPC: cast CuAl post and core; SSP: stainless steel post + composite core; GP: fiberglass + composite core; CP: carbon fiber + composite core; ZP: zirconium dioxide post + composite core; TP: titanium post + composite core. The numerical models were considered to be restored with a leucite-reinforced all ceramic crown and received a 45 masculine occlusal load (10 N) on the lingual surface.All the materials and structures were considered linear elastic, homogeneous, and isotropic, with the exception of fiberglass and carbon fiber posts which assumed orthotropic behavior. The numerical models were plotted and meshed with isoparametric elements, and the results were analyzed using von Mises and Sy stress criteria. When compared with the sound tooth, FEA revealed differences in stress distribution when post systems were used. Among the restored teeth, the use of CPC, SSP, ZP, and TP resulted in higher stress concentration in the post itself when compared to GP and CP. Therefore, results from the FEA images suggested that the use of non-metallic post systems could result in improved mechanical behavior for the weakened restored teeth. PMID- 20019418 TI - Effect of filler content of flowable composites on resin-cavity interface. AB - The purpose of this in vitro study was to evaluate marginal integrity and wall adaptation in 1- and 2-mm-deep cavities restored with a high filler-loaded flowable composite in comparison to a flowable composite with lower filler content and a conventional hybrid composite. 1-mm-deep dentin and 2-mm-deep enamel-dentin Class I cavities were prepared and restored with a self-etch adhesive and with one of the composites. Samples were crosscut and evaluated for marginal integrity and gap formation using a digital microscope. Selected samples were also observed using a scanning electron microscope. For 1-mm-deep cavities, no differences in marginal integrity and wall adaptation were observed among the three composites. For 2-mm-deep cavities, those restored with the bulk technique and with the higher filler-loaded flowable composite demonstrated a similar outcome as that of the conventional hybrid composite. On the effect of restorative techniques, cohesive enamel defects were observed in bulk-filled 2-mm deep cavities. However, when the incremental technique was used in conjunction with the higher filler-loaded flowable composite and the conventional hybrid composite for 2-mm-deep cavities, superior results were obtained. PMID- 20019419 TI - Effects of plasma treatment on the shear bond strength between fiber-reinforced composite posts and resin composite for core build-up. AB - The aim of this study was to evaluate the effects of plasma treatment on adhesion between fiber-reinforced posts and a composite core material. Two types of posts, methacrylate-based (FRC Postec) and epoxy resin-based (DT Light-Post), were treated with oxygen plasma (O(2)), argon plasma (Ar), nitrogen plasma (N(2)), or helium mixed with nitrogen plasma (He+N(2)) using a radio-frequency generator before bonding to a methacrylate-based composite. Pull-out tests were performed using a universal testing machine. Surface roughness of each group was evaluated using a profilometer. On tensile-shear bond strength, statistical analysis revealed that the type of post, type of plasma treatment, and their interaction significantly influenced the results (p<0.05). Tukey's test revealed significant differences in tensile-shear bond strength between the control and other plasma treatment groups (p<0.05). On surface roughness, Tukey's test revealed significant differences between the control group and the Ar group (p<0.05) with DT Light Post. Plasma treatment appeared to increase the tensile-shear bond strength between post and composite. PMID- 20019420 TI - Effects of light sources and visible light-activated titanium dioxide photocatalyst on bleaching. AB - The objective of this study was to evaluate, using methylene blue (MB), the effects of various light sources on the bleaching action of hydrogen peroxide (H(2)O(2)) with two titanium dioxide (TiO(2)) photocatalysts - an ultraviolet light-activated TiO(2) photocatalyst (UVTiO(2)) versus a visible light-activated TiO(2) photocatalyst (VL-TiO(2)). Five experimental solutions (VL TiO(2)+H(2)O(2), UV-TiO(2)+H(2)O(2), H(2)O(2), VL-TiO(2), UV-TiO(2)) were prepared by mixing varying concentrations of H(2)O(2 )and/or TiO(2 )photocatalyst with MB solution. For H(2)O(2)-containing solutions (VL-TiO(2)+H(2)O(2), UV TiO(2)+H(2)O(2), and H(2)O(2)), the concentration of H(2)O(2) was adjusted to 3.5%. For the four different light sources, low- and high-intensity halogen lamps and blue LED LCUs were used. All the experimental solutions were irradiated by each of the light sources for 7 minutes, and the absorbance at 660 nm was measured every 30 seconds to determine the concentration of MB as an indicator of the bleaching effect. On the interaction between the effects of light source and bleaching treatment, the high-intensity halogen with VL-TiO(2)+H(2)O(2) caused the most significant reduction in MB concentration. On the effect of light sources, the halogen lamps resulted in a greater bleaching effect than the blue LED LCUs. PMID- 20019421 TI - Adhesion of hollow calcium-deficient hydroxyapatite microspheres onto titanium. AB - Microcapsules containing active drugs, e.g. an antimicrobial agent, with a hydroxyapatite (HAp) shell are expected to prevent infection and to improve osseointegration simultaneously when used as implant materials. This study was conducted to investigate the effects of etching with hydrofluoric acid (HF) to change the surface morphology of titanium in cases of adhesion of calcium deficient HAp microspheres onto titanium. The microspheres were mixed with Hanks' balanced salt solution (HBSS) to prepare slurry; the slurry was then put between two titanium disks that had been etched and kept soaking in HBSS for seven days at 310 K. The coverage and the degree of adherence of the microsphere were evaluated using electron probe microanalysis. The etching at an HF concentration of 0.10 mol/L caused the greatest adhesion. Changing the temperature between 303 and 323 K showed a tendency for the degree of adherence to increase. PMID- 20019422 TI - Resistance of nanofill and nanohybrid resin composites to toothbrush abrasion with calcium carbonate slurry. AB - The aim of this study was to investigate the wear of four nanofilled resin composites using simulated toothbrushing for 50,000 cycles with calcium carbonate slurry. The depth of abrasion and roughness (Ra) were measured after each 10,000 brushing cycle. The surface texture of the worn samples was examined by SEM.The wear depths of the nanofill Filtek Supreme XT (FIL), the nanohybrides Grandio (GRA), Tetric EvoCeram (TET), and Venus Diamond (VED) increased linearly with numbers of brushing cycles or approximately 80, 12, 600, and 60 mum, respectively after 50,000 strokes. Surface roughness showed virtually no change between 10,000 and 50,000 brushing cycles; the ranking order was TET < FIL < GRA < VED. FIL showed rather uniform abrasion with nanoclusters protruding from the surface. TET was very smoothly abraded without signs of debonding of the prepolymerized particles, whereas GRA and VED showed pronounced wear of the matrix polymer surrounding larger glass filler particles. PMID- 20019423 TI - The acid-base resistant zone in three dentin bonding systems. AB - An acid-base resistant zone has been found to exist after acid-base challenge adjacent to the hybrid layer using SEM. The aim of this study was to examine the acid-base resistant zone using three different bonding systems. Dentin disks were applied with three different bonding systems, and then a resin composite was light-cured to make dentin disk sandwiches. After acid-base challenge, the polished surfaces were observed using SEM. For both one- and two-step self etching primer systems, an acid-base resistant zone was clearly observed adjacent to the hybrid layer - but with differing appearances. For the wet bonding system, the presence of an acid-base resistant zone was unclear. This was because the self-etching primer systems etched the dentin surface mildly, such that the remaining mineral phase of dentin and the bonding agent yielded clear acid-base resistant zones. In conclusion, the acid-base resistant zone was clearly observed when self-etching primer systems were used, but not so for the wet bonding system. PMID- 20019424 TI - Prediction of the reduced glutathione (GSH) reactivity of dental methacrylate monomers using NMR spectra - Relationship between toxicity and GSH reactivity. AB - It has been established that the toxicity of acrylate and methacrylate monomers is driven by their reactivity towards glutathione (GSH). With this relationship, the objective of this study was to predict the GSH reactivity of dental methacrylate monomers, and hence their toxicity, using the (13)C-NMR chemical shifts of beta-carbon (delta(Cbeta)) and the (1)H-NMR shifts of the protons attached to beta-carbon (delta(Ha), delta(Hb)). The different nucleophiles were chosen to compare the different nucleophilic reactions involving acrylate and methacrylate monomers. In conjunction with the use of literature data for monomer/GSH reactivity, significant linear relationships between GSH reactivity (log K) and delta(Cbeta )or delta(Ha )were observed (p<0.001). As for the oral LD(50 )values of some dental dimethacrylates in mice, they were estimated using linear regression curve fitting of GSH reactivity-toxicity response data. Results revealed an acceptable correlation between the oral LD(50 )values of acrylates and methacrylates and GSH reactivity (p<0.05, outlier: HEMA). In conclusion, the present findings suggested that NMR spectra might be useful for predicting the toxicity of dental methacrylates. PMID- 20019425 TI - Influence of flowable resins on the shear bond strength of orthodontic brackets. AB - The present study tested the shear bond strength (SBS) of orthodontic brackets bonded to teeth using flowable resins. A total of 105 human teeth were divided into seven groups. The brackets in each group were bonded to the tooth substrates using a Transbond XT adhesive and six other different flowable resins, respectively. Each adhesive resin was light-cured using a quartz-tungsten-halogen (QTH) light for 40 seconds. The brackets were debonded in shear mode using a universal testing machine. The brackets bonded using Tetric Flow and Grandio Flow showed slightly greater SBS values (13.0+/-1.8 and 12.2+/-1.2 MPa, respectively) than the Transbond XT adhesive (12.1+/-1.2 MPa). However, the SBS values of Transbond XT, X-flow, Tetric Flow, Grandio Flow, and Filtek Z350 flow were not significantly different. As for Admira Flow and Aelite Flow, they scored significantly lower SBS values (7.0+/-2.1 and 9.2+/-1.3 MPa, respectively) than the other groups. On bracket failure mode, statistical analysis revealed a similar failure mode among all the seven test groups. PMID- 20019426 TI - Surface modification of stainless steel by plasma-based fluorine and silver dual ion implantation and deposition. AB - The aims of this study were to modify dental device surface with fluorine and silver and to examine the effectiveness of this new surface modification method. Stainless steel plates were modified by plasma-based fluorine and silver ion implantation-deposition method. The surface characteristics and brushing abrasion resistance were evaluated by XPS, contact angle and brushing abrasion test. XPS spectra of modified specimens showed the peaks of fluoride and silver. These peaks were detected even after brushing abrasion test. Water contact angle significantly increased due to implantation-deposition of both fluorine and silver ions. Moreover, the contact angle of the modified specimen was significantly higher than that of fluorine only deposited specimen with the same number of brushing strokes. This study indicates that this new surface modification method of fluorine and silver ion implantation-deposition improved the brushing abrasion resistance and hydrophobic property making it a potential antimicrobial device. PMID- 20019428 TI - [Investigation of a phantom for diffusion weighted imaging that controlled the apparent diffusion coefficient using gelatin and sucrose]. AB - When studying diffusion weighted imaging(DWI), it is important to create a phantom that has a reliably controlled diffusion coefficient. In this study, we investigated phantoms to control both the diffusion coefficient and the T2-value by changing the concentration of gelatin or sucrose and MnCl(2), respectively. The results showed that the diffusion coefficient decreased linearly with increases in the gelatin or sucrose concentration, and decreasing of their relaxation times was observed. By properly adjusting the MnCl(2) concentrations, we were able to equalize the T2-values between phantoms having different gelatin or sucrose concentrations. Temperature dependence of the diffusion coefficient was also revealed. This phantom can be made stable for a few months by adding a small amount of NaN(3) as an antiseptic agent, has a diffusion coefficient similar to that of neural tissue or clinical tumor, and is able to control the T2 value properly. We consider this phantom suitable for studying SE-type DWI and contributes to elucidation of this technique. PMID- 20019427 TI - Dentin bond strength of a new adhesive system containing calcium phosphate experimentally developed for direct pulp capping. AB - The purpose of this study was to evaluate the microtensile bond strength (microTBS) to human dentin of an experimental bonding agent containing calcium phosphates experimentally developed for direct pulp capping. Different concentrations of four types of calcium phosphates were added to an experimental bonding monomer, and six experimental bonding agents were thus prepared. Clearfil SE Bond/Bond was used as the control. Flat dentin surfaces of human molars were assigned to the experimental adhesive systems and the control. After Clearfil SE Bond/Primer was applied to the dentin surface, each experimental bonding agent was applied and photopolymerized, and then a resin composite paste was placed and photopolymerized. The specimens were subjected to microTBS testing. Results revealed that there were no significant differences among the microTBS values of the experimental bonding agents and the control. In other words, the calcium phosphate-containing experimental adhesives did not adversely affect the microTBS to dentin. PMID- 20019429 TI - [Distortion in diffusion weighted imaging]. AB - The definitional equation of the distortion of echo planar imaging(EPI)was examined. We compared measured values with calculated values by using the definitional equation of the chemical shift of the EPI method that first composed the diffusion-weighted image, and examined the possibility of applying it to the distortion. The results showed that the chemical shift with the definitional equation and the measurement corresponded, and the correlation between the chemical shift and the distortion was acquired. Next, the distortion of each images that composed DWI with the increase and the sign acceptable method difference of b value. The difference in the distortion between each image has increased with the increase in b value. It was assumed that the influence of the eddy currents was due to the high motion probing gradient. PMID- 20019430 TI - [Examination of the means of measuring liver function in the hepatobiliary phase]. AB - In a field of contrast-enhanced magnetic resonance imaging of the liver, attention has been focused on evaluation of liver function using gadolinium ethoxybenzyl diethylenetriaminepentaacetic acid(EOB). In this study, we examined the possibility of obtaining liver function in only one hepatobiliary phase 60 minutes after injection. First, in regard to the difference between the signal intensity of two materials, we examined the effects of slice gap, surface coil intensity correction(SCIC), and others. Secondly, we compared the difference between liver and spleen signal intensity with biochemical laboratory tests, Child-Pugh class, liver damage class, and the two indices(HH(15) and LHL(15))calculated by 99mTc-DTPA-galactosyl-human serum albumin hepatic scintigraphy in patients with chronic liver diseases. Finally, we designated the "Liver EOB uptake index(L-EOB(60))" from those results, compared with HH(15) and LHL(15). The results demonstrated that the difference between the signal intensity of two materials increased in the lack of slice gap explained by cross talk, and decreased with SCIC. The difference between liver and spleen signal intensity decreased with worsened liver and kidney function. In the case of slice gap >20% and direct bilirubin <0.5 mg/dL without SCIC, the correlation coefficient between L-EOB(60) and LHL(15) was 0.97. L-EOB(60) was strongly proportional to LHL(15). We conclude that L-EOB(60) meeting the above conditions can be employed as a useful index to determine liver function. PMID- 20019431 TI - [Effect of ambient light variations on luminance ratio of CRT monitor and image contrast in ultrasonography]. AB - Variation in the luminance ratio of a cathode ray tube(CRT)monitor and the ultrasonographic images at different levels of ambient light(0-150 lux)was investigated to obtain optimum ambient light in the ultrasonography suite. The maximum and minimum luminances of test patterns and ultrasonographic images were measured after three technicians independently optimized the brightness and contrast of the CRT monitor and ultrasonographic images at different levels of ambient light. Furthermore, the luminance ratio was calculated from the maximum luminance divided by the minimum luminance. When ambient light increased, it was difficult for the technicians to optimize the brightness and contrast settings of the CRT monitor to maintain a high luminance ratio at 0 lux. The luminance ratio decreased rapidly as ambient light increased up to 20 lux. However, the luminance ratio decreased gradually when ambient light was higher than 20 lux. It is necessary to take into consideration the ambient light to maintain a high luminance ratio of ultrasonographic images. PMID- 20019432 TI - [A study of ADC value in PROPELLER DWI]. AB - Diffusion weighted imaging(DWI)is rapidly becoming widespread in MRI, with many reports of the differential diagnosis using the Apparent Diffusion Coefficient(ADC)being seen. We examined the influence of scan parameter on ADC in PROPELLER-DWI. Neither the TR nor T1 value had a major influence on the ADC value. Number of MPG pulse, effective TE, and T2 value were the factors in which the ADC value was changed. In addition, the ADC value has been changed by the scan sequence. The factor that influenced the ADC value was able to be confirmed by this experiment. It is necessary to understand the influence that an individual factor has on ADC. PMID- 20019433 TI - [Investigation of actual conditions of mammography in Kagoshima Prefecture]. AB - We surveyed the actual conditions of mammography with regard to image quality and radiation dose at 44 facilities in Kagoshima prefecture in 1999. In April 2004, guidelines for mammography newly included the standard of digital mammography. From September to October 2005, the survey was conducted at 48 facilities, and the results of the survey were compared with that in 1999. We visited 44 of the 48 facilities, and visually evaluated the image quality of mammograms for RMI156 and clinical mammograms. In addition, we measured average mammary gland dose at each facility. The number of the mammography device that satisfied the specified guideline criterion was larger than that in 1999. Image quality for the RMI156 mammograms improved. However, the results of the present survey revealed several problems. First, the number of facilities that had quality control instruments for mammography are few. Second, radiological technologists, medical doctors, and nurses did not share knowledge or information regarding mammography. Finally, there were differences in devices and image quality for mammography among the facilities. We achieved an understanding of the actual conditions of mammography in Kagoshima prefecture by visiting many facilities, evaluating image quality, and communicating with many staff members. Our results may be useful for the development of mammography examinations. PMID- 20019434 TI - [How to introduce film-less operation]. PMID- 20019435 TI - [The strategy to the construction of electric medical records systems--hopes and effects from the Department of Radiology]. PMID- 20019437 TI - [Modulation transfer function--presampled MTF measurements by Fourier analysis using square wave chart]. PMID- 20019436 TI - [Structure and control of medical linear accelerator]. PMID- 20019438 TI - [Fundamental of medical image processing with personal computer system--image J is used. Chapter of DR image]. PMID- 20019439 TI - [Teleradiology]. PMID- 20019440 TI - [The interim report No. 2 of investigative research on patient exposure doses in diagnostic X-ray examinations]. PMID- 20019442 TI - [Medical electrical equipment--Part 1-3: General requirements for basic safety and essential performance--collateral standard: radiation protection in diagnostic X-ray equipment JIS T 0601-1-3]. PMID- 20019443 TI - [For the economical value evaluation of radiological technology & medical device]. PMID- 20019444 TI - Involvement of ATP-sensitive K(+) channels in the peripheral antinociceptive effect induced by the alpha(2)-adrenoceptor agonist xylazine. AB - Xylazine is an alpha(2)-adrenergic agonist extensively used in veterinary medicine and animal experimentation for producing antinociception, sedation, and muscle relaxation. The nitric oxide (NO) / cGMP / ATP-sensitive K(+) (K(ATP)) channel pathway has been proposed as the action mechanism of peripheral antinociception of several groups of drugs, including opioids and nonsteroidal analgesics. Considering the lack of knowledge regarding the mechanisms involved in xylazine effects, the present study investigated the contribution of K(+) channels on peripheral antinociception induced by xylazine using the rat paw pressure test, in which hyperalgesia was induced by intraplantar injection of prostaglandin E(2). Xylazine administered into the right hind paw elicited a local antinociceptive effect, since only much higher doses produced a systemic effect in the contralateral paw. The peripheral antinociceptive effect induced by xylazine was antagonized by glibenclamide, a specific blocker of K(ATP) channels. In another experiment, tetraethylammonium, a voltage-dependent K(+)-channel blocker, and paxilline and dequalinium, which are selective blockers for the large- and small-conductance Ca(2+)-activated K(+) channels, respectively, were ineffective at blocking xylazine antinociception. These results provide evidence that the peripheral antinociceptive effect of xylazine probably results from K(ATP)-channel activation, while the voltage-dependent K(+) channels, small- and large-conductance Ca(2+)-activated K(+) channels, appear not to be involved in this mechanism. PMID- 20019445 TI - Exendin-4 potentiates insulinotropic action partly via increasing beta-cell proliferation and neogenesis and decreasing apoptosis in association with the attenuation of endoplasmic reticulum stress in islets of diabetic rats. AB - Exendin-4, a long-acting glucagon-like peptide-1-receptor agonist, is known to enhance beta-cell function, but the active mechanism by which it modulates beta cell mass still remains unclear. We investigated what the long-term effects of exendin-4 (300 pmol/kg body weight per day) on beta-cell function and mass would be in 90% pancreatectomized (Px) Sprague Dawley rats; half of whom were intraperitoneally injected with streptozotocin (STZ, 20 mg/kg body weight) and half of whom were not. Exendin-4 improved glucose tolerance by elevating serum insulin levels in both STZ-treated and untreated Px rats. At hyperglycemic clamp, STZ attenuated both first and second phase insulin secretion in STZ- and saline treated Px rats, but exendin-4 incompletely reversed the attenuation. Since STZ mostly removed the remaining beta-cells by increasing apoptosis after Px, their regeneration was initiated through neogenesis, which was determined by the number of beta-cells budding from pancreatic duct layers and small clusters. Exendin-4 enhanced beta-cell proliferation and neogenesis in STZ-treated and -untreated Px rats and reduced beta-cell apoptosis partly by attenuating the expression of endoplasmic reticulum stress-response genes such as X-box-binding protein-1, activating transcription factor (ATF)-4, ATF6, and C/EBP-homologous protein. In conclusion, exendin-4 improved glycemic control by potentiating beta-cell function and increasing beta-cell mass by increasing beta-cell neogenesis and proliferation and by decreasing apoptosis in diabetic rats. PMID- 20019446 TI - Alterations in dystrophin-related glycoproteins in development of right ventricular failure in rats. AB - Genetic depletion of the dystrophin-related glycoprotein (DRGP) complex causes cardiomyopathy in animals and humans. The present study was undertaken to explore the possible involvement of alterations in DRGP in the development of the right ventricular failure in monocrotaline-administered rats (MCT rats). At the 6th and 8th weeks after subcutaneous administration of 60 mg/kg monocrotaline, echocardiographic examination showed that cardiac output indices were decreased and that the right ventricular Tei indices were increased, suggesting that right ventricular failure occurs, at the latest, by 6 weeks after monocrotaline administration. The levels of alpha- and beta-sarcoglycan and beta-dystroglycan in the right ventricle of the MCT rats at the 6th and 8th weeks were markedly decreased, and these decreases were inversely related to the increase in the right ventricular Tei index of the MCT-administered animals. The content and activity of the Ca(2+)-activated neutral protease m-calpain in the right ventricle of the MCT rats were increased at the 4th to 8th weeks and those of matrix metalloproteinase-2, at the 6th and 8th weeks. These results suggest that m-calpain- and/or matrix metalloproteinase-2-mediated alterations in the contents of alpha-sarcoglycan, beta-sarcoglycan, and beta-dystroglycan may be involved in the development of right ventricular failure in MCT rats. PMID- 20019447 TI - The distinct roles of calmodulin and calmodulin kinase II in the reversal of run down of L-type Ca(2+) channels in guinea-pig ventricular myocytes. AB - In this study, we investigated the roles of calmodulin kinase II (CaMKII) and calmodulin (CaM) in the reversal of run-down of L-type Ca(2+) channels. Single Ca(2+)-channel activities in guinea-pig ventricular myocytes were recorded using the patch-clamp technique, and run-down of the channel activities was induced by inside-out patch formation in the basic internal solution. At 1 min after patch excision, 1 - 30 muM CaMKII mutant T286D (CaMKIIT286D), a constitutively active type of CaMKII, induced the Ca(2+)-channel activities to only 2% - 10% of that recorded in the cell-attached mode. However, in the presence of CaMKIIT286D, the time-dependent attenuation of CaM's effects in the reversal of run-down was abolished. A GST-fusion protein containing amino acids 1509 - 1789 of the C terminal region of guinea-pig Cav1.2 (CT1) was prepared. In pull-down assays, CT1 treated with CaMKIIT286D showed a higher affinity for CaM compared with CT1 treated with phosphatase. We propose a model in which CaMKII-mediated phosphorylation of the channels regulates the binding of CaM to the channels in the reversal of run-down of L-type Ca(2+) channels. PMID- 20019448 TI - Developmental changes in hepatic antioxidant capacity are age-and sex-dependent. AB - Developmental inadequacy in hepatic antioxidant defenses may contribute to chemical toxicity and pediatric liver diseases. We measured a comprehensive panel of antioxidants in liver tissue from 27 normal pediatric donors. Glutathione reductase declined with age (P = 0.008, r = -0.54, Spearman) while microsomal glutathione-S-transferase increased (GST, P<0.001, r = 0.81). Males had significantly lower superoxide dismutase and vitamin E (P<0.05) and may have lower glutathione reductase (P = 0.06), while females show less cytosolic GST (P = 0.07). Hepatic antioxidants are high in neonates, decline throughout childhood, and then increase in adolescence to adult levels. PMID- 20019449 TI - The value of real-time three-dimensional transesophageal echocardiography in the assessment of paravalvular leak origin following prosthetic mitral valve replacement. AB - OBJECTIVES: Two-dimensional (2D) echocardiographic approaches are not sufficient to determine the origin of paravalvular leak (PVL) that occurs after prosthetic mitral valve replacement (MVR). In this study, we investigated the role of real time three-dimensional transesophageal echocardiography (RT-3D TEE) in detecting the origin and size of PVL occurring after prosthetic MVR. STUDY DESIGN: The study included 13 patients (7 females; 6 males; mean age 56+/-10 years; range 37 to 71 years) who developed PVL within a mean of 8.3+/-3.8 years following mechanical prosthetic MVR. Nine patients (69.2%) had atrial fibrillation, and four patients (30.8%) had normal sinus rhythm. Four patients (30.8%) had hemolysis. Paravalvular leak was mild, moderate, and severe in two, six, and five patients, respectively. Real-time 3D TEE was performed using a 3D matrix-array TEE transducer immediately after detection of PVL on 2D TEE examination. Localization of PVL was made using a clock-wise format in relation to the aortic valve and the size of dehiscence was measured. RESULTS: The mean PVL width measured by 2D TEE was 3.00+/-0.92 mm. The mean length of dehiscence was 13.6+/ 8.8 mm, and the mean width was 3.88+/-2.04 mm on RT-3D TEE. The PVLs were mainly localized in the posterior and anterior annular positions between 12 to 03 hours (n=7) and 06 to 09 hours (n=3) on RT-3D TEE, respectively, which corresponded to the posteromedial or anterolateral sectors of the posterior annulus. CONCLUSION: Considering that only the width of the PVL defect can be assessed by 2D TEE, delineation by RT-3D TEE includes the localization of PVL together with the length and width of the defect. PMID- 20019450 TI - [Two years of multidisciplinary diagnostic and therapeutic experience in patients with pulmonary arterial hypertension]. AB - OBJECTIVES: Information is limited on the prognosis of patients with pulmonary arterial hypertension (PAH) in Turkey. We evaluated our multidisciplinary diagnostic and therapeutic experience in PAH patients. STUDY DESIGN: The study included 51 patients (32 women, 19 men; mean age 45.4+/-9.7 years) who were prospectively monitored during a two-year period by the PAH Working Group in our hospital. The diagnoses were as follows: idiopathic/familial PAH (n=9); PAH associated with connective tissue disease (n=16), congenital heart disease (n=11), and with pulmonary veno-occlusive disease (n=1); chronic thromboembolic pulmonary hypertension (n=10), and other causes (n=4). The patients were assessed every three months with clinical examination, six-minute walk test, transthoracic echocardiography, and BNP levels. RESULTS: The mean pulmonary artery pressure was 54.7+/-18.8 mmHg. Functional capacity was NYHA class II in nine patients (17.7%), class III in 28 patients (54.9%), and class IV in 14 patients (27.5%). Thirty seven patients (72.6%) received treatment with specific pharmacological agents, in whom 19 patients (51.4%) required modifications during treatment. Nine patients (17.7%) benefited from treatment with decreases of at least one NYHA class, whereas NYHA class remained unchanged in 25 patients (49%). Seventeen patients (33.3%) exhibited clinical deterioration, of whom 11 died with an overall mortality of 21.6%. Patients who died were all in NYHA class III or IV and significantly differed from those who survived with respect to mean pulmonary artery pressure (72.5+/-18.7 mmHg vs. 49.8+/-21.2 mmHg), BNP level at the time of diagnosis (293.8+/-88.3 pg/ml vs. 141.6+/-62.1 pg/ml), and six-minute walk distance (123.8+/-41.3 m vs. 200.7+/-52.1 m) (p<0.05). CONCLUSION: Despite relative improvements in the end points over the last two decades, PAH is detected late in the course of the disease, resulting in severe functional and hemodynamic problems in the majority of patients. PMID- 20019451 TI - [Evaluation of serum adiponectin levels in patients with heart failure and relationship with functional capacity]. AB - OBJECTIVES: We aimed to evaluate serum adiponectin levels in relation to the NYHA functional capacity class in patients with heart failure (HF). STUDY DESIGN: The study included 49 patients (40 males, 9 females; mean age 63 years) with HF, whose functional capacity was NYHA class II to IV. Echocardiographic examination was performed and serum adiponectin levels were measured. The results were compared in relation to the NYHA classes and with those of 41 control subjects (24 males, 17 females; mean age 54.2 years) without HF. RESULTS: Functional capacity was NYHA class II in 13 patients (26.5%), class III in 23 patients (46.9%), and class IV in 13 patients (26.5%). Compared to the control group, the HF group exhibited a significantly higher mean age (p=0.001), lower body mass index (p=0.004), decreased left ventricular ejection fraction (EF) (33.2+/-7.7% vs. 64.9+/-4.3%; p=0.0001), and increased serum adiponectin level (4.0+/-3.2 micromg/dl vs. 2.4+/-2.3 micromg/dl; p=0.009). Both EF (p=0.001) and adiponectin level (p=0.004) showed significant differences between the NYHA groups, with the latter showing a sharp increase from 2.6+/-2.6 micromg/dl in class II to 6.8+/ 3.7 micromg/dl in class IV. In all paired comparisons between the three NYHA groups, EF and serum adiponectin level exhibited significant differences except for the serum adiponectin level for NYHA class II and III (for NYHA class II and IV, p=0.003; for class III and IV, p=0.008). In correlation analysis, serum adiponectin level was in a significantly inverse correlation with EF (r=-0.380, p=0.0001), and a positive correlation with the NYHA class (r=0.423, p=0.0001). CONCLUSION: Serum adiponectin levels significantly increase in patients with HF, in parallel with deterioration in functional capacity and with significant decreases in EF. PMID- 20019452 TI - Decreased coronary flow reserve in obese women. AB - OBJECTIVES: Obesity is associated with an increased rate of cardiovascular disease and risk factors. It is a common problem in apparently healthy women. We aimed to investigate the association between obesity and coronary flow reserve (CFR) in obese women. STUDY DESIGN: The study included 80 consecutive women (mean age 55.6+/-10.2 years) without diabetes mellitus and clinical coronary artery disease. Body mass index (BMI) was calculated and obesity was defined as BMI = or >30 kg/m(2). Based on BMI, the patients were grouped as normal weight (n=13; 18.5 24.9 kg/m(2)), overweight (n=32; 25-29.9 kg/m(2)), obese (n=32; = or >30-39.9 kg/m(2)), and morbid obese (n=3; = or > 40 kg/m(2)). Peak diastolic coronary flow velocities were measured in the distal left anterior descending artery by transthoracic pulsed wave Doppler echocardiography at baseline and after dipyridamole infusion and CFR was calculated as the ratio of hyperemic to baseline peak diastolic velocities. RESULTS: There were 35 obese women (43.8%). Coronary flow reserve was significantly lower in obese women than in nonobese subjects (2.2+/-0.5 vs. 2.5+/-0.4; p=0.022). The lowest CFR was seen in patients with a BMI of = or > 40 kg/m(2); overweight women did not differ significantly from women of normal weight. Coronary flow reserve was correlated with BMI (r= 0.314, p=0.005), waist circumference (r=-0.316, p=0.005), C-reactive protein (CRP) (r=-0.342, p=0.011), and adiponectin level (r=0.410, p=0.011). In regression analysis, BMI (p=0.017), waist circumference (p=0.048), systolic blood pressure (p=0.025), fasting glucose (p=0.035), and adiponectin level (p=0.037) were found to be independent predictors for impaired CFR. In ROC analysis, the cut-off value for BMI to predict impaired CFR was = or > 30 kg/m(2), with 76% sensitivity and 72% specificity (ROC area 0.805, p<0.001, 95% CI 0.669-0.96). CONCLUSION: Impaired CFR in obese women suggests the presence of microvascular dysfunction. Treatment of obesity is important for the prevention of atherosclerosis. PMID- 20019453 TI - The relationship between coronary calcification and the metabolic markers of osteopontin, fetuin-A, and visfatin. AB - OBJECTIVES: We investigated whether coronary calcification detected by multislice computed tomography (MSCT) was correlated with plasma osteopontin, serum fetuin A, and visfatin levels. STUDY DESIGN: The study included 64 consecutive patients (51 males, 13 females; mean age 49.5+/-10.9 years; range 33 to 78 years) who underwent MSCT for suspected coronary artery disease. Coronary artery calcification (CAC) scores of the patients were calculated using the Agatston scoring method. Plasma osteopontin, serum fetuin-A, and visfatin levels were measured from fasting blood samples and correlations were sought with calcium scores. RESULTS: Coronary calcification was detected in 32 patients (50%). The mean CAC score was 146.5+/-333.7 Agatston units (AU), indicating an intermediate risk for coronary artery disease. In 10 patients (15.6%), the CAC score exceeded 400 AU. The mean fetuin-A, visfatin, and osteopontin levels were 25.6+/-6.4 ng/ml, 19.7+/-47.2 ng/ml, and 20.4+/-16.1 ng/ml, respectively. Serum visfatin (r=0.15, p=0.37) and fetuin-A (r=0.17, p=0.22) were not correlated with the CAC score, whereas plasma osteopontin level showed a moderate correlation with the CAC score (r=0.35; p=0.008). In ROC analysis, the area under the curve for identification of CAC was greatest for osteopontin (0.741; p=0.004), followed by fetuin-A (0.574; p=0.31), and visfatin (0.580; p=0.27). The cut-off value was 18.45 ng/ml for osteopontin, with a sensitivity of 72% and specificity of 73%. CONCLUSION: Our results suggest that there might be an association between CAC and plasma osteopontin levels. Research should continue to find out a metabolic parameter that will strongly indicate coronary calcification. PMID- 20019454 TI - An unusual cause of OptiVol alarm: increased intra-abdominal pressure associated with irritable bowel syndrome. AB - Monitoring intrathoracic impedance has become an integral part of follow-up of patients with implantable cardioverter defibrillator (ICD) or cardiac resynchronization therapy/defibrillator due to heart failure. However, several noncardiac factors may influence intrathoracic impedance. We report on an unusual cause of decrease in intrathoracic impedance in a 54-year-old male patient following successful implantation of biventricular ICD for heart failure symptoms due to nonischemic dilated cardiomyopathy and severely impaired left ventricular systolic function. During the follow-up period, the patient presented several times with the OptiVol alarm due to an increase in the OptiVol fluid index, in the absence of symptoms or signs of heart failure. Further inquiry into the possible causes of decreased intrathoracic impedance revealed that the patient had frequent episodes of irritable bowel syndrome, which increased intra abdominal pressure, leading to elevation of diaphragm and subsequent compression of intrathoracic organs, and thus to a decrease in intrathoracic impedance. PMID- 20019455 TI - Late detection of noncompaction of the myocardium in an adult with complete interventricular septal defect. AB - Noncompaction of the ventricular myocardium (NVM) is a morphogenetic anomaly that leads to the development of cardiomyopathy. It is often associated with other congenital cardiac malformations. Common clinical presentations of NVM involve heart failure symptoms, ventricular tachyarrhythmias, and thromboembolic events. Although the peculiar echocardiographic picture is characteristic for this entity, it may often be misdiagnosed. In this case report, we describe a 27-year old man who had been followed-up since childhood, with the diagnosis of interventricular septal defect or enlarged "single ventricle" with a very small rudiment of the apical part of the interventricular septum. On his last echocardiographic examination, NVM was detected with heavy trabeculations and intertrabecular recesses. This case suggests that physicians should be more patient to detect other congenital abnormalities including NVM, which may influence the clinical and prognostic outcome of these patients. PMID- 20019456 TI - Acute intoxication with propafenone and trimethoprim-sulfamethoxazole in a case of suicide attempt. AB - A 17-year-old male ingested about 20 tablets of propafenone (total 6,000 mg) and 24 tablets of trimethoprim (total 1,920 mg)--sulfamethoxazole (total 9,600 mg) with suicidal intent. Within one hour, he was brought to a hospital with vomiting, nausea, and loss of consciousness, where he developed cyanosis and mild acidosis, and eventually cardiorespiratory arrest, despite bicarbonate, saline infusion, and inotropic support. Fortunately, he was fully resuscitated and ventilated, and sinus rhythm was restored. He was then transported to our center. On admission, his heart rate was regular with 55 beats/min and blood pressure was 70/45 mmHg. The 12-lead electrocardiogram (ECG) showed sinus bradycardia, extreme widening of the QRS complex (260 msec) with a right bundle branch block pattern. Intravenous saline, bicarbonate, and dopamine were administered, and respiration was supported mechanically, which resulted in rapid restoration of sinus rhythm and improvement in hemodynamic parameters and acidosis. A subsequent ECG showed shortening of the QRS duration (230 msec). He was discharged with an appropriate hemodynamic balance on the third day with normal ECG findings. PMID- 20019457 TI - Endovascular stenting for treatment of superior vena cava syndrome. AB - With the advances in interventional cardiology, percutaneous treatment by stenting has become a reasonable strategy in superior vena cava syndrome (SVCS), whether the underlying disease is malignant or benign. We present a 48-year-old woman with typical signs and symptoms of SVCS, who was treated with endovascular stenting by the percutaneous approach. We obtained both procedural success with complete restoration of blood flow and immediate relief of symptoms. During a follow-up period of six months, the patient was free of symptoms and computed tomography demonstrated complete stent patency. PMID- 20019458 TI - [Endomyocardial fibrosis causing right intraventricular gradient and obliteration]. AB - Endomyocardial fibrosis is a cause of restrictive cardiomyopathy and it generally occurs in tropical regions more commonly affecting children and young adults. A 19-year-old male patient presented with edema in the lower extremities and fatigue. Transthoracic echocardiography showed dilated right heart chambers, restrictive physiology in the left ventricle, and increased tissue growth in the right ventricle that caused a 60-mmHg gradient and obliteration. Magnetic resonance imaging confirmed the presence of increased tissue formation in the right ventricular inflow region. Surgical resection was not considered taking into account the functional capacity of the patient (class II), disappearance of symptoms following medical treatment, and the high risk for operative mortality. The presented case may arouse interest in that increased tissue growth in the right ventricle inflow region caused a gradient in the right ventricle, leading to an incorrect diagnosis, at another center, as idiopathic pulmonary hypertension. PMID- 20019459 TI - Mediastinal lymphoma causing extrinsic pulmonary stenosis. AB - Acquired pulmonary stenosis is rare in adults and may be missed unless a high index of suspicion is present. Extrinsic pulmonic stenosis is even rarer and predominantly caused by external thoracic masses creating non-dynamic obstruction of the right ventricular outflow tract. A 20-year-old female was referred to our center with a cystic mass detected by transthoracic echocardiography and thoracic computed tomography, in the superoanterior mediastinum, 5.5 x 5.5 x 7 cm in size, causing main pulmonary trunk compression. Repeat transthoracic echocardiography demonstrated a mass causing compression of the main pulmonary artery immediately after the pulmonary valve. Continuous wave Doppler showed a peak systolic gradient of 65 mmHg and a mean gradient of 37 mmHg in the pulmonary artery at the site of compression. She underwent an open thoracotomy via a midline sternotomy. The mass was firmly attached to the pericardium. Its largest diameter was 15 cm; it surrounded the left phrenic nerve completely and invaded the outer wall of the pulmonary artery and aorta. The mass could only be partly dissected. The pathological diagnosis of the mass was stage IIa nodular sclerosing Hodgkin's lymphoma. The patient received postoperative chemotherapy and thoracal radiotherapy. She was in remission without any cardiac complaint. PMID- 20019460 TI - [Major influence of dysfunctions of protective serum proteins on cardiometabolic risk among Turks and gender difference]. AB - Knowledge obtained from the Turkish Adult Risk Factor (TARF) study on higher morbidity and mortality rates compared to other populations from coronary heart disease (CHD) among Turkish adults has been confirmed recently with greater power. This review provides insight that the dysfunctions of the protective serum proteins, attaining pro-inflammatory and atherogenic features, may be attributed to atherogenic dyslipidemia, oxidative stress, and systemic inflammation associated with the high prevalence of metabolic syndrome (MetS) among Turks. The mentioned protective protein dysfunctions, firstly described in a general population to date, are high-density lipoprotein (HDL), apolipoprotein (apo) A-I, A-II, and apoC-III, apart from adiponectin. Based on published findings of the TARF study, this review discusses the role of inflammatory mediators such as elevated C-reactive protein (CRP), apoB, apoC-III, fibrinogen, and low adiponectin serum levels in cardiometabolic risk comprising MetS, type 2 diabetes, and CHD, the degree of independence of these mediators from the ATP-III defined MetS, and the influence of sex. Moreover, it is emphasized that dysfunctions of adiponectin and protective proteins related to HDL particles increase not only cardiometabolic risk significantly but also CHD risk among half of Turkish adults in a magnitude similar to or greater than that associated with traditional risk factors. Also underlined is the observation that cigarette smoking reduces the risk in Turkish women for the development of hypertension, MetS, and diabetes by mediation of positive effects on dysfunctional apoA-I, visceral fat accumulation and, above all, CRP levels. This knowledge is of utmost importance and sheds light to authorities and those concerned on the necessity of urgent and radical modifications regarding strategies in prevention and management of cardiovascular health of middle-aged Turks. PMID- 20019461 TI - Quadricuspid aortic valve diagnosed by transthoracic echocardiography. PMID- 20019462 TI - Pseudo-myocardial infarction pattern in a patient with spontaneous pneumomediastinum. PMID- 20019463 TI - Demonstration of apical hypertrophic cardiomyopathy by left ventriculography and computed tomographic angiography. PMID- 20019464 TI - [Hydatid cyst in the left ventricle]. PMID- 20019465 TI - [The diagnostic value of N-terminal B-type natriuretic peptide in diastolic heart failure: comparison with echocardiographic findings]. PMID- 20019466 TI - Editorial: A different approach to cardiovascular risk factors. PMID- 20019467 TI - [Renin-angiotensin-aldosterone system inhibition and cardiovascular protection]. AB - The primary aim of the treatment in hypertension is to prevent cardiovascular complications. All hypertensives reduce the risk for cardiovascular events by providing effective blood pressure control. Besides blood pressure-lowering effect, the contribution of an antihypertensive agent to reduce the risk for cardiovascular events at high-risk patients was first demonstrated in 2000 in the HOPE study (Heart Outcomes Prevention Evaluation), which used an angiotensin converting enzyme (ACE) inhibitor, ramipril 10 mg (N Engl J Med 2000;342:145-53). However, at the time the results of this study appeared, the use of angiotensin receptor blockers (ARB) was gaining popularity in the treatment of hypertension as an alternative to ACE inhibitors. This raised the question as to whether an ARB would also offer benefit comparable to that derived from ramipril. A non inferiority trial was planned by the same research group to test whether telmisartan 80 mg was as effective as ramipril 10 mg in a similar patient population. In order to obtain reliable results, 25,000 patients from all over the world had to be followed-up for five years (Am Heart J 2004;148:52-61). The results of the ONTARGET study (The Ongoing Telmisartan Alone and in Combination with Ramipril Global Endpoint Trial) which were presented at the annual meeting of the American College of Cardiology in March 2008 were suggestive of a new indication for telmisartan. In August, 2009, the U.S. Food and Drug Administration reported that a new indication might be justified for the use of telmisartan, an angiotensin-receptor blocker: In order to reduce the risk for cardiovascular diseases and if an ACE inhibitor (ramipril 10 mg) cannot be used, telmisartan 80 mg can be used in patients with a high cardiovascular risk profile (Press announcements, FDA). The aim of this review is to provide a comprehensive analysis of the course of this recent development in cardiovascular protection. PMID- 20019468 TI - Control of hypertension in Turkey--is it improving? The Kocaeli 2 study. AB - OBJECTIVES: In a previous study conducted in Kocaeli, Turkey, ten years before, a very low rate of hypertension control (6.2%) was reported. This hypertension survey was conducted to determine whether the treatment and control rates of hypertension improved in the same region. STUDY DESIGN: Using a stratified sampling method, 1222 subjects (559 males, 663 females, mean age 44.8+/-13.6 years) living in Kocaeli, aged 25 or over, were included. After administration of a standardized questionnaire, blood pressure was measured in the sitting position from both arms after at least five minutes of resting. The measurement was repeated after 10 minutes on the arm with the higher initial reading. Hypertension was defined as the presence of a mean systolic blood pressure = or >140 mmHg and/or a mean diastolic blood pressure = or >90 mmHg, or a previous diagnosis of hypertension and/or use of antihypertensive drugs. RESULTS: The prevalence of hypertension was 33.6% (37.1% in women, and 29.5% in men, p<0.001). Of the hypertensive subjects, 59.3% were aware of their condition. Hypertension was controlled in 8.7% of the subjects (systolic blood pressure <140 mmHg and diastolic blood pressure <90 mmHg). CONCLUSION: Our results showed that about one third of the adult population in Kocaeli had hypertension, with low rates of control and awareness. Since the previous survey, there has been a small improvement in hypertension control rate, but uncontrolled hypertension is still remarkably high in this geographical region. These findings are consistent with the recent Turkish Hypertension Prevalence study (PatenT) reporting a control rate of 8.1%. PMID- 20019469 TI - Cut-off values for waist circumference in Turkish population: Is there a threshold to predict insulin resistance? AB - OBJECTIVES: To identify the cut-off values for waist circumference (WC) that may best predict cardiometabolic risk associated with insulin resistance in adult Turkish men and women. STUDY DESIGN: A total of 1,039 non-diabetic subjects (592 females, 447 males; mean age 40.2+/-12.8 years) aged = or >18 years were enrolled. Insulin resistance was assessed with homeostasis model assessment of insulin resistance (HOMA-IR). The subjects were classified into quantiles based on the WC values and quantiles were compared with respect to the HOMA-IR levels. Receiver operating characteristic (ROC) analysis was used to identify the WC cut off value that would best predict insulin resistance. RESULTS: HOMA-IR values increased in parallel with increasing WC quantiles for both genders (p=0.001). The cut-off values for WC that showed the highest sensitivity and specificity to predict insulin resistance were 93 cm for men (91.6% and 51%, respectively) and 83 cm for women (91.1% and 34.8%, respectively). CONCLUSION: Our results demonstrate that the cut-off values for WC that would best predict cardiometabolic risk associated with insulin resistance are 93 cm for men and 83 cm for women in Turkish adults. However, our findings also raise the question as to whether it is indeed necessary to define a WC cut-off value for the prediction of insulin resistance, because, regardless of the cut-off WC value taken into account, insulin resistance will be overlooked in a significant number of individuals with a WC below this cut-off value. PMID- 20019470 TI - [Unmet needs in hypertension]. AB - Hypertension is considered one of the world's most important health problems. Approximately one billion people are affected in the world. Almost for 50 years, effective medical treatment in reducing blood pressure are among the tools of cardiovascular medical treatment. However, control rates of hypertension is not at an acceptable level. One of the main causes of the difficulty to take hypertension under control is that this disease is a chronic disease. Control of hypertension and the goal of optimum blood pressure is of crucial importance because of the association between arterial hypertension and organ damage in terms of cardiac, vascular and renal functions. An important part of research on hypertension has concentrated on renin angiotensin system (RAS), as a basic mechanism in regulating acute and chronic blood pressure. Today, what is expected from new antihypertensive drugs and direct renin inhibitors in particular is to provide significantly more effective blood pressure control as well as preventing target organ damage, by acting on the RAS which has an important role in the etiology of hypertension. PMID- 20019471 TI - [What is renin inhibition? Mechanism of action]. AB - After discovery of the first direct renin inhibitor, aliskiren, which blocks the renin-angiotensin-aldosterone system in the first rate limiting step, in addition to angiotensin-converting enzymes (ACE) and angiotensin-receptor blockers (ARB), renin has become an important target nowadays. The scope of this review is to give a detailed information regarding renin which has an important regulatory function in the body. It is crucial to understand renin and related structures in order to understand its functions in the organ systems. It is for sure that the ongoing trials will enlighten us more regarding the beneficial effects of renin inhibition in terms of morbidity and mortality. PMID- 20019472 TI - [Renin inhibition in hypertension]. AB - Aliskiren is a novel molecule which blocks the renin-angiotensin-aldosterone system in its rate limiting step by renin inhibition. Aliskiren is validated as an antihypertensive drug in many countries based on its efficacy data. Although some surrogate endpoint trials suggest that its potential advantages based on its mechanism of action provide clinical benefit beyond its blood pressure-lowering effect, hard endpoint clinical trials to reveal its long-term effects are yet to be completed. PMID- 20019473 TI - [Renin and the heart]. AB - The inhibition of renin-angiotensin system (RAS) has an effect beyond reducing blood pressure in the treatment of cardiovascular diseases; it also reduces mortality and morbidity. Although the classic RAS blockage is effective, due to blockage of negative feedback inhibition, it increases plasma renin activity and as a result generates a residual cardiovascular risk. Different from ACE-i and ARB treatments, aliskiren, a direct renin inhibitor, is the only pharmacologic agent that noticeably reduces the plasma renin activity, an independent cardiovascular risk factor. This creates a new option in RAS blockage. The efficacy and safety of aliskiren in the treatment of hypertension has been well established. The effect of aliskiren on cardiovascular mortality and morbidity is being researched. At this point there are many experimental and clinical studies to answer questions on this subject. PMID- 20019474 TI - [Renin inhibition and the kidney]. AB - Renin-angiotensin aldosterone system has a detrimental effect on the progression of renal failure in patients with chronic kidney disease. Drugs inhibiting the renin-angiotensin aldosterone system, such as angiotensin-converting enzyme inhibitors and angiotensin-receptor blockers have been shown to slow the progression of kidney disease in patients with diabetic and nondiabetic renal diseases. Aliskiren is a direct renin inhibitor that blocks the renin-angiotensin system at its initial step. Several experimental studies have demonstrated the renoprotective effects of aliskiren. Aliskiren, added to standard therapy, shows additional and significant antiproteinuric effects in patients with diabetic nephropathy. The optimal therapeutic approach to patients with target organ damage will be better understood as the ongoing large clinical studies with direct renin inhibitors conclude. PMID- 20019475 TI - [The future of renin inhibition]. AB - Although antihypertensive drugs currently used provide significant decreases in blood pressure and improve clinical results, cardiovascular morbidity and mortality are not sufficiently decreased; therefore, there is still a need for new approaches to the treatment of hypertension and related cardiovascular diseases. However, when a new blood-pressure lowering therapy is introduced, the question of whether this will be superior over other drug classes in terms of its advantages in hypertensive patients is frequently asked. In 1898, Tigerstedt and Bergman discovered "renin" as a consequence of an observation of blood-pressure elevation following the injection of rabbit renal extracts to rabbits; however, the first member of the renin system pharmacology, ACE inhibitors, could be developed in 1970's. This was followed by the development of angiotensin-receptor blockers (ARB) and, during the last 30 years, it has been shown that the pharmacologic blockage of renin-angiotensin system (RAS) improves the prognosis in hypertensive patients. It has been shown that renin system is the key system in the treatment of hypertension and related comorbidities and that the drugs which target renin system, such as ACE inhibitors and ARB, reduce the cardiovascular events to a large extent. In contrast, as the inhibition of angiotensin II (Ang II) production and effect prevents the negative feedback which helps Ang II to inhibit the renin release from the kidney, elevated Ang II levels suggest that renin enzyme, which can be considered to be the center of renin system, can be the optimal tool in the treatment. Aliskiren, which is the first oral direct renin inhibitor developed based on these ideas, was approved by the FDA in March 2007. During all this period, clinical studies have shown that aliskiren is as efficient as other antihypertensive drugs, and preclinical studies have shown that, in genetically modified rats, aliskiren is efficient in healing the cardiovascular damage associated with Ang II. The fact that the plasma renin activity (PRA), which increases with other antihypertensive therapies and is associated with increased cardiovascular complications, decreases with the use of direct renin inhibitors raises the question as to whether aliskiren may provide additional benefit in reducing cardiovascular morbidity and mortality. The ASPIRE HIGHER program designed to find an answer to this question includes several studies which investigate how aliskiren impacts the clinical results. The studies AVOID, ALOFT and ALLAY, which were completed within the ASPIRE HIGHER program, showed that aliskiren had beneficial effects for surrogate markers of cardiovascular and renal diseases, while AGELESS study showed that, in elderly with systolic hypertension, aliskiren (150 or 300 mg) provided a higher blood pressure lowering compared to ramipril (5 or 10 mg). In addition, many studies included in this program are ongoing and the results of these studies will provide more information about direct renin inhibition. Potential effects of RAS in cognitive functions and the functions of its different components such as angiotensin and AT4 receptor are the topics which are still awaiting answers. PMID- 20019503 TI - Perspectives on critical care education. Foreword. PMID- 20019504 TI - Assessing and developing critical-thinking skills in the intensive care unit. AB - A lot of resources are spent on the development of new staff in the intensive care unit (ICU). These resources are necessary because the environment in the ICU is complex and the patients are critically ill. Nurses need an advanced knowledge base, the ability to accurately define and change priorities rapidly, good communication and teamwork skills, and the ability to work in a stressful environment in order to succeed and give their patients quality care. Critical thinking helps the nurse to navigate the complex and stressful environment of the ICU. Critical thinking includes more than just nursing knowledge. It includes the ability to think through complex, multifaceted problems to anticipate needs, recognize potential and actual complications, and to expertly communicate with the team. A nurse who is able to think critically will give better patient care. Various strategies can be used to develop critical thinking in ICU nurses. Nurse leaders are encouraged to support the development of critical-thinking skills in less experienced staff with the goal of improving the nurse's ability to work in the ICU and improving patient outcomes. PMID- 20019505 TI - The role of preceptor and nurse leader in developing intensive care unit competency. AB - In the past, only experienced nurses were hired to work in critical care areas. With the prolonged nursing shortage, more and more facilities are hiring recent nursing graduates to work in these fast paced critical care units. How does this impact the orientation process in critical care? What competencies and skills are necessary in a nurse that will be working in a critical care setting? Are there specific competencies all critical care nurses must possess, even those with little or no previous experience? How do you help new registered nurses develop critical-thinking skills during orientation? These are very important questions that need to be addressed in order to get the outcome we want by the end of orientation: a competent critical care registered nurse. This article will explore significant factors that have the capacity to negatively or positively influence the success of an orientation program. During my years of nursing practice, I have found 4 factors that are especially crucial to the success of a critical care orientation program. These factors include: 1. strong leadership support throughout the comprehensive orientation process. 2. appropriate preceptor selection. 3. focus on developing critical-thinking skills. 4. use of a competency-based orientation plan. PMID- 20019506 TI - Evidence-based teaching guidelines: transforming knowledge into practice for better outcomes in healthcare. AB - Educational change is necessary to meet the demands of the current healthcare environment. An outcome-based approach to healthcare education is optimal to support organizational change. Learning objectives should focus on the best practice outcomes and should emphasize what the healthcare provider is expected to do after the educational activity is over. Regulating agencies and consumers hold healthcare accountable for providing high-quality, safe patient care. Educational activities should provide the skills and knowledge that enable nurses to meet this goal. PMID- 20019508 TI - Journey to Oz: the yellow brick road to a blended learning environment. AB - The 21st century has ushered in many new trends and tools in education. A myriad of articles, Web sites, and books offer enticing possibilities that Web 2.0 tools may increase learner engagement, improve interactivity, and allow tailoring of education to the learner. Classroom training is trending downward, and Web-based training is on the rise. But is the Internet really the answer to educators' prayers, or is it just another flash in the pan? The sheer number of technological tools and delivery methods available can be mind-boggling. This article reviews some of the basic terminology and tools that are readily available for use in delivering education via the Internet. Benefits and barriers associated with e-learning are discussed from the evidence found in nursing and education literature and the personal experiences of a nurse educator in a multihospital system. PMID- 20019509 TI - Use of simulation training in the intensive care unit. AB - With increased patient acuity, nurse educators are faced with the challenge of providing ongoing education and ensuring nurse competency. High-fidelity simulation is an effective teaching modality that can meet the diverse needs of critical care nurses with various backgrounds and levels of experience. The Institute of Medicine recommends the use of simulation and simulators as a best practice tool for an instructional method in healthcare. There is documented support for the integration of simulation in nursing continuing education. Organizations such as the Agency for Healthcare Research and Quality, American College of Surgeons, American Council for Graduate Medical Education, National League for Nursing, American Nursing Association, and American Association of Critical Care Nurses have provided support and encouragement for the use of innovative teaching modalities to create enhanced learning. Along with enhanced learning, simulation plays a role in improving patient safety, recruitment and retention strategies, and addressing quality and risk-management concerns. This article will focus on high-fidelity simulation as it relates to clinical education and nurse competency in the intensive care unit. PMID- 20019510 TI - Certification and clinical ladder as the impetus for professional development. AB - With today's healthcare challenges of nursing shortages and financial instability, it is imperative that healthcare organizations retain clinically competent nurses at the bedside. Professional development and recognition are key motivators to increase nursing job satisfaction, thus reducing shortages and turnover. Implementation of specialty certification and clinical advancement programs is of benefit to the public, employers, and nurses alike. Clinical ladder and Magnet recognition are often the impetus for specialty nursing certification in healthcare institutions. Clinical ladder history, purpose, models, perceptions, and satisfiers are discussed. Certification statistics, types, impetus, benefits, incentives, and barriers are highlighted, as well as a facility's innovative strategy to increase specialty certification. Certification and clinical ladder programs demonstrate commitment of healthcare organizations and nursing staff to provide high-quality care and professional nursing development, an investment that hospitals cannot afford to overlook. PMID- 20019511 TI - The benefits and challenges of providing nursing student clinical rotations in the intensive care unit. AB - The goal of providing a clinical rotation in a basic nursing program is to integrate skills and knowledge from the classroom setting into the clinical practice setting. In the intensive care unit (ICU), nursing students have the ability to learn about the complex health issues of critically ill patients, practice selected technical skills, and develop communication skills. There are both benefits and challenges to having nursing students in the intensive care setting. With preparation, the student is able to immerse in the ICU environment, acquire new knowledge and skills, and participate alongside the nurse caring for critically ill patients. The staff nurse must balance patient care with the added responsibilities of helping the student meet the clinical goals. It is optimal to have faculty that are also intensive care clinically competent and can facilitate the clinical experience. The school, the hospital, and the ICU need to collaborate to provide a positive clinical experience that is safe for the patient. In return, the hospital can recruit student nurses and clinical faculty. Planned with thought and intention, rotations in the ICU can be an ideal clinical setting for upper-level student nurses to learn the role of the registered nurse. PMID- 20019512 TI - Generational considerations in providing critical care education. AB - With the current and predicted nursing shortage, much emphasis is placed on recruitment and retention. With an aging workforce, we must recruit, educate, and retain nurses from many different generations. As leaders and educators, we must be aware of generational differences and work with staff to appreciate potential preferences in communication, approach to learning and motivational factors. We are aware that over the next 15 years, many experienced nurses will retire. We must do all we can to recruit and retain nurses from all generations in order to provide a workforce able to meet the needs of our patients and families. Generational preferences should be considered when developing nursing education and in welcoming and accepting new staff into the culture of the nursing unit. PMID- 20019513 TI - Sleep deprivation among critical care patients. AB - To gain an understanding and increased knowledge regarding the presence and affects of ambient stressors on patients' sleep and the efficacy of implementing sleep promotion measures within the intensive care unit environment, the author reviewed 10 empirical studies. Research indicates that ambient stressors within the intensive care unit have detrimental effects on patients' sleep, and nursing interventions that focus on the abatement of ambient stressors enhance patients' sleep. In the intensive care unit, optimizing the environment to promote sleep requires the active removal of ambient stressors. PMID- 20019514 TI - A discussion of family-centered care within the pediatric intensive care unit. AB - Every year, thousands of children are admitted to pediatric intensive care for treatment. Many of these admissions are for acute injuries, but children with chronic illnesses requiring repeated hospitalization are also on the rise. Hospitalization of a child is extremely stressful for both the patient and family. Historically, intensive care units had restrictive visitation hours and did not allow for sibling visitation or multiple family members. Parents and family members were not encouraged to participate in care when at the bedside. As the shift toward family-centered care continues, many hospitals are now changing visitation policies to allow for active family involvement in patient care. Parents are now encouraged to participate in care. Intensive care units are modifying layouts of the unit to facilitate visitors and provide sleeping spaces for parents when available. Families are considered part of the team instead of visitors, and are included in the decision making process. The purpose of this article is to promote discussion of family-centered care in the pediatric intensive care unit. PMID- 20019515 TI - How do critical care nurses define a "good death" in the intensive care unit? AB - The purpose of this qualitative study was to explore and describe the characteristics of a good death as defined by 15 critical care nurses working in the intensive care unit (ICU) at a hospital in a mid-sized urban city. The target population was registered nurses employed in the intensive care unit where the study was conducted. Given the fact that the sample population was a very specific group, a purposive, convenience sample was utilized to explore the personal thoughts and feelings of the nurses who volunteered for this project. A nondirective, in-depth interview technique was the method of data collection. Interviews lasted up to 60 minutes, and data was collected by tape recorder, then immediately transcribed verbatim; basic demographic data was collected at the beginning of each interview and this information was used to describe the sample. In describing a good death in the ICU, most participants identified multiple themes. Eight main themes emerged from the nurses' responses to the research question, and of those, 3 were mentioned most often. The 3 most frequently mentioned themes were patient does not die alone, patient does not suffer (pain management/symptom management), and acceptance of death by the patient and/or loved ones. Much of what nurses described as elements of a good death in the ICU supports other published results. The themes that were identified in this research study provide an initial framework that is important for further research in the area of critical care nursing. PMID- 20019561 TI - Clinical validation study of percutaneous cochlear access using patient customized microstereotactic frames. AB - OBJECTIVE: Percutaneous cochlear implant (PCI) surgery consists of drilling a single trough from the lateral cranium to the cochlea avoiding vital anatomy. To accomplish PCI, we use a patient-customized microstereotactic frame, which we call a "microtable" because it consists of a small tabletop sitting on legs. The orientation of the legs controls the alignment of the tabletop such that it is perpendicular to a specified trajectory. STUDY DESIGN: Prospective. SETTING: Tertiary referral center. PATIENTS: Thirteen patients (18 ears) undergoing traditional cochlear implant surgery. INTERVENTIONS: With institutional review board approval, each patient had 3 fiducial markers implanted in bone surrounding the ear. Temporal bone computed tomographic scans were obtained, and the markers were localized, as was vital anatomy. A linear trajectory from the lateral cranium through the facial recess to the cochlea was planned. A microtable was fabricated to follow the specified trajectory. MAIN OUTCOME MEASURES: After mastoidectomy and posterior tympanotomy, accuracy of trajectories was validated by mounting the microtables on the bone-implanted markers and then passing sham drill bits across the facial recess to the cochlea. The distance from the drill to vital anatomy was measured. RESULTS: Microtables were constructed on a computer-numeric-control milling machine in less than 5 minutes each. Successful access across the facial recess to the cochlea was achieved in all 18 cases. The mean +/- SD distance was 1.20 +/- 0.36 mm from midportion of the drill to the facial nerve and 1.25 +/- 0.33 mm from the chorda tympani. CONCLUSION: These results demonstrate the feasibility of PCI access using customized microstereotactic frames. PMID- 20019562 TI - Bone-anchored hearing aid: a comparison of surgical techniques. AB - OBJECTIVE: To determine which bone-anchored hearing aid (BAHA) implantation surgical technique is associated with the fewest major postoperative complications and shortest time between surgery and use of the BAHA. The techniques evaluated were 1) a free retroauricular "full-thickness" skin graft, 2) a pedicled parieto-occipital epidermal graft, 3) a dermatome-pedicled parieto occipital dermal graft, and 4) two broad pedicled local epidermal skin envelopes/skin flaps. STUDY DESIGN: Retrospective case study. SETTING: Tertiary referral center. PATIENTS: One hundred forty-three patients who received a BAHA at Maastricht University Medical Center between November 1996 and January 2007. Number and mean age of patients in each group: Technique 1 (n = 30; mean age, 55 yr), Technique 2 (n = 45; mean age, 54 yr), Technique 3 (n = 47; mean age, 55 yr), and Technique 4 (n = 21; mean age, 54 yr). MAIN OUTCOME MEASURES: Cumulative proportion of implants that remained free of major complications versus follow-up interval, time between surgery and use of BAHA. RESULTS: Technique 4 (2 broad pedicled local epidermal envelopes/skin flaps) has a significantly higher proportion of implants that remained free of major complications during first year of follow-up (91%; p = 0.021). Pairwise comparisons revealed that Technique 4 also has a significantly shorter time until use (2 mo) than Techniques 1 (2.5 mo), 2, and 3 (both 2.3 mo). CONCLUSION: Two broad pedicled, local epidermal envelopes/flaps are associated with significantly fewer major complications and have one of the shortest times between surgery and use of the BAHA. The use of a dermatome is not associated with fewer major complications. We recommend Technique 4 as the preferred standard in BAHA surgery to minimize complications, postoperative medication, discomfort, and cost. PMID- 20019563 TI - Response to "Riechelmann H, Tholen M, Keck T, Rettinger G. Perioperative glucocorticoid treatment does not influence early post-laser stapedotomy hearing thresholds. Am J Otol 2000;21:809-812". PMID- 20019564 TI - Dominant dorsal duct syndrome: a rare cause of acute recurrent pancreatitis in children revisited. AB - Acute recurrent pancreatitis in children can be caused by anomalies of fusion of pancreatic ducts such as the dominant dorsal duct syndrome wherein a dominant dorsal pancreatic duct is associated with stenosis of the minor papilla. Clinical presentations and management of 2 patients are discussed. An infant presented with severe acute pancreatitis with pseudocyst formation due to an underlying ductal disruption. Surgical treatment was offered on account of failure of medical therapy and endoscopic stenting. A dominant dorsal duct with minor papilla stenosis was encountered. Sphincteroplasty of the minor papilla and lateral pancreaticojejunostomy were performed with good result. A 14-year-old boy with a type 1 choledochal cyst was troubled by recurrent acute pancreatitis. At operation, a dilated dorsal pancreatic duct opening into a stenosed minor papilla was found in addition to the choledochal cyst. Choledochal cyst excision, choledochoduodenostomy, and sphincteroplasty of the minor papilla stenosis were performed. Dominant dorsal duct syndrome is a rare cause of acute pancreatitis in children. A high index of suspicion is necessary to establish a precise diagnosis. Sphincteroplasty of the minor papilla may affect adequate pancreatic drainage and prevent recurrent pancreatitis. PMID- 20019565 TI - An unusual variant of anomalous pancreaticobiliary junction in a patient with pancreas divisum diagnosed with secretin-magnetic resonance cholangiopancreatography. AB - Anomalous pancreaticobiliary junction is an unusual variant of pancreaticobiliary anatomy of clinical importance because it is associated with increased risk of pancreatitis and, also, for the development of cholangiocarcinoma. We report an unusual variant of anomalous pancreaticobiliary junction, occurring in a patient with pancreas divisum, with an anomalous communication between the dorsal pancreatic and the common bile duct. The patient presented with a distal biliary stricture. This variant anatomy was occult on magnetic resonance cholangiopancreatography but was diagnosed on magnetic resonance cholangiopancreatography that was performed with intravenous secretin administration and further delineated by endoscopic retrograde cholangiopancreatography. The features that allowed the diagnosis to be made and the implications of this diagnosis are described in this report. PMID- 20019566 TI - Importance of quantitative analysis in the generation of insulin-expressing cells from human embryonic stem cells. PMID- 20019567 TI - Serum and urine trypsinogen activation peptide in assessing post-endoscopic retrograde cholangiopancreatography pancreatitis. PMID- 20019568 TI - First case of 2 intraductal papillary mucinous tumors of both ventral and dorsal ducts in pancreas divisum. PMID- 20019569 TI - Total pancreatectomy for intractable pain in chronic pancreatitis? PMID- 20019570 TI - Invading and long-lasting enormous pancreatic head tumor. PMID- 20019571 TI - Early predictive factors of in hospital mortality in patients with severe acute pancreatitis. PMID- 20019572 TI - Successful single-balloon enteroscopic dilation of late anastomotic pancreaticojejunostomy stricture following whipple procedure. PMID- 20019573 TI - Nicotine gum causing pancreatitis: a case report. PMID- 20019574 TI - High prevalence of symptomatic acute HIV infection in an outpatient ward in southern Mozambique: identification and follow-up. AB - OBJECTIVES: To determine the prevalence of acute HIV infection (AHI) within the HIV-seronegative adult population presenting with reported fever in a district hospital in southern Mozambique and evaluate clinical, immunological and virological parameters of AHI. DESIGN: This is a prospective observational study. METHODS: Three hundred and forty-six adults presenting with reported fever at an outpatient ward at the Manhica District Hospital in Mozambique were screened for AHI by HIV rapid serology testing, followed by HIV-RNA testing in HIV seronegative individuals. Plasma from HIV-seronegative patients was pooled in the ratio of 1: 5 for HIV-RNA testing. Whole blood was used for Plasmodium falciparum rapid test determination at screening visit. Follow-up visits at day 7, 4 and 10 months included clinical examination, HIV serotesting and assessment of HIV-RNA, CD4 cell counts and percentage of activated CD8 T cells. RESULTS: HIV serotesting revealed that 37.8% (95% confidence interval 32.7-43.2) of the adults had previously undiagnosed established HIV infection. Among the HIV-seronegative patients, 3.3% (95% confidence interval 1.3-6.7) were found to have AHI as demonstrated by positive HIV-1 RNA testing. Median HIV-1 RNA levels at diagnosis of AHI were 6.21 log10 copies/ml (interquartile range 5.92-6.41) and significantly higher than median HIV-RNA load at 4 months. At day 7 after screening, patients showed a median CD4 cell count of 384 cells/microl (interquartile range 239-441) and a median percentage of activated CD8 T cells of 68.4% (interquartile range 59.6-87.8). CONCLUSION: Of patients reporting with fever, 3.3% were shown to be potentially due to AHI. High prevalence of AHI in southern African populations may warrant investigation of tools and target populations for AHI screening as a novel way to address HIV prevention. PMID- 20019575 TI - Kidney function and the risk of cardiovascular events in HIV-1-infected patients. AB - OBJECTIVE: Cardiovascular events (CVEs) are a significant cause of mortality in HIV/AIDS patients. The objective is to determine the correlation between kidney function and the risk of CVEs in the HIV-infected population. DESIGN: Nested, matched, case-control study design was employed. METHODS: : We performed a single center study of 315 HIV-infected patients (63 patients who had CVEs and 252 controls). Estimated glomerular filtration rate (eGFR), calculated by the Chronic Kidney Disease Epidemiology Collaboration formula and the Modification of Diet in Renal Disease equation, and proteinuria were the primary exposures of interest. RESULTS: Mean eGFR was significantly lower in the patients compared with controls (68.4 vs. 103.2 ml/min per 1.73 m, P < 0.001 by Chronic Kidney Disease Epidemiology Collaboration formula and 69.0 vs. 103.1 ml/min per 1.73 m, P < 0.001 by Modification of Diet in Renal Disease equation). In univariate analysis, an eGFR of less than 60 ml/min per 1.73 m was associated with a 15.9-fold increased odds of a CVE compared with an eGFR of at least 60 ml/min per 1.73 m (P < 0.001). In multivariate analysis, a 10 ml/min per 1.73 m decrease in eGFR was associated with a 20% increased odds of a CVE (odds ratio 1.2, 95% confidence interval 1.1-1.4). The prevalence of proteinuria in the patients was approximately twice that of controls (51 vs. 25%, P < 0.001). Proteinuria was associated with CVEs both in univariate and multivariate analyses (odds ratio 3.6, 95% confidence interval 1.9-7.0 and odds ratio 2.2, 95% confidence interval 1.1-4.8, respectively). Traditional cardiovascular risk factors, such as history of previous CVEs, diabetes mellitus, and dyslipidemia, along with low CD4 cell counts were also found as significant predictors of risk of CVEs. CONCLUSION: Our study shows a significant independent association between decreased kidney function and increased risk of CVE in HIV-1-infected patients. PMID- 20019576 TI - Ensuring sustainable antiretroviral provision during economic crises. PMID- 20019578 TI - Palliative effects of an incomplete cytoreduction combined with perioperative intraperitoneal chemotherapy. AB - OBJECTIVES: Incomplete cytoreduction often occurs because of overwhelming burden of disease intraperitoneally that limit a successful cytoreduction. The outcome of an incomplete cytoreduction followed by administering perioperative intraperitoneal chemotherapy on whether it prolongs survival or palliates symptoms is yet to be established. METHODS: A retrospective review of a prospectively collected database was performed. Eleven patients were identified to have had an incomplete (CC2/3) cytoreduction and received either hyperthermic intraperitoneal chemotherapy or early postoperative intraperitoneal chemotherapy. The symptoms of the patients before and after treatment during follow-up consultations in clinic were noted. Survival analysis was performed using the Kaplan Meier method. RESULTS: The median follow-up period was 19 (range, 0.5-35) months. The overall median survival was 21 months with a 1- and 2-year survival rate of 72% and 36%, respectively. Five of 7 patients with pseudomyxoma peritonei and 2 of 3 patients with colorectal cancer peritoneal carcinomatosis experienced symptom improvement after treatment. Severe postoperative morbidity and achieving symptomatic improvement following treatment seems to be associated with a dismal prognosis. CONCLUSION: Our results suggest that some patients derive a survival and symptomatic benefits despite an incomplete cytoreductive surgery and perioperative intraperitoneal chemotherapy. The effects of this treatment require further investigation to determine its benefits as a palliative procedure. PMID- 20019577 TI - A phase II trial of imatinib mesylate in merkel cell carcinoma (neuroendocrine carcinoma of the skin): A Southwest Oncology Group study (S0331). AB - BACKGROUND: Imatinib mesylate (Gleevec) was evaluated as a treatment for Merkel cell carcinoma (MCC, neuroendocrine carcinoma of the skin) based on the identification of strong c-KIT staining of these neoplasms. METHODS: Eligibility included patients with measurable metastatic or unresectable MCC, c-KIT (CD117) expression and a Zubrod performance status of 0 to 2. Imatinib 400 mg daily was administered orally in 28-day cycles to 23 patients. RESULTS: Overall, imatinib was well tolerated with grade 1 or 2 nausea, diarrhea, and hematologic toxicity as the most frequent side effects. A partial response was seen in 1 patient (4%; 95% CI: 0%-22%). Median progression-free survival was 1 month (95% CI: 1-2 months). Median overall survival was 5 months (95% CI: 2-8 months). One patient achieved a partial response and another had prolonged disease stabilization while receiving treatment. CONCLUSIONS: The majority of patients progressed rapidly within 1 to 2 cycles of treatment. The observed progression-free survival and overall survival were not adequate to conclude that this agent was active in advanced MCC, and thus the planned second stage of patient accrual was not opened. PMID- 20019579 TI - Generalized tetanus despite prior vaccination and a protective level of anti tetanus antibodies. AB - Because of the success of widespread vaccination, tetanus rarely occurs in developed countries such as the United States. Vaccination, however, is not entirely protective even if patients develop an adequate antibody response. We discuss a case of generalized tetanus in an individual with both a history of prior vaccination and a measurable level of anti-tetanus antibodies at the time of presentation. We speculate that the patient's preexisting antibodies may have moderated his course of illness. PMID- 20019580 TI - A 73-year-old-man with fever and weight loss. PMID- 20019581 TI - Dose-response and self-efficacy effects of an exercise program on vigor change in obese women. AB - BACKGROUND: The causative factors of exercise-induced mood change remain unclear. The objective of this study was to clarify the relationship of exercise with change in vigor (energy level). METHOD: Obese women in an exercise support treatment (n = 122) or control condition (n = 70) were assessed on vigor, exercise barriers self-efficacy, and physical self-concept at baseline and at month 6. Exercise session attendance was also assessed in the treatment group. The relationship between exercise attendance and vigor change was tested. Multiple regression analyses also tested whether change in vigor was predicted by exercise barriers self-efficacy and physical self-concept scores, both at baseline and change over 6 months. RESULTS: Vigor, exercise barriers self efficacy, and physical self-concept were significantly improved in the treatment group only. Exercise session attendance was not significantly related to change in vigor. Multiple regression analyses indicated that exercise barriers self efficacy and physical self-concept scores, both at baseline and change from baseline to month 6, explained a significant portion of the variance in changes in vigor (Radj = 0.13 and 0.21, respectively). In post hoc analyses, entry of depressed mood (indicated by a cut point on depression score) into the regression equations increased the variance accounted for to Radj = 0.15 and 0.25, respectively. CONCLUSIONS: Findings supported behavioral explanations of the exercise-mood change relationship emanating from social cognitive and self efficacy theory. The use of manageable amounts of exercise early in the weight loss intervention process was suggested. PMID- 20019582 TI - Extramedullary-intradural spinal metastasis of small cell lung cancer causing cauda equina syndrome. AB - Small cell lung cancer (SCLC) has a very aggressive clinical progression with widespread metastases. We describe the case of a patient with SCLC treated with concurrent chemoradiotherapy. One month later, after finishing the scheduled treatment, the patient was admitted to the hospital again with symptoms of low back pain that radiated to bilateral lower legs with painful paresthesias, urinary incontinence, and constipation. After a series of examinations, including bone scan and magnetic resonance imaging, the patient received an L2-L3 laminectomy. The concluded diagnosis through histopathologic examination with immunohistochemistry was extramedullary-intradural spinal metastasis causing cauda equina syndrome. The metastatic tumour originated from the SCLC. To the best of our knowledge, this is the first reported case of SCLC metastasized to the cauda equina causing cauda equina syndrome. PMID- 20019583 TI - Case report of human immunodeficiency virus infection, Hodgkin lymphoma, and pregnancy. AB - As the incidence of human immunodeficiency virus (HIV) infection in women of child bearing age continues to increase in the era of highly active antiretroviral therapy and Hodgkin lymphoma (HL) is the most common non-acquired immunodeficiency syndrome defining malignancy, we anticipate that the number of cases of HIV-associated HL in pregnant women will increase in the near future. Herein, we describe the case of a pregnant 30-year-old HIV-infected Ethiopian woman with a CD4 count of 254 cells/microL and an HIV viral load of 1200 copies/mL who presented to medical attention with progressive neck adenopathy. Subsequent histopathology and radiographic findings revealed clinical stage IIIA Classical HL. After a spontaneous miscarriage of 10 weeks into her pregnancy, the patient began highly active antiretroviral therapy and chemotherapy. Thirty months later, she remained in complete remission. Through a literature review, we identified 2 additional case reports involving HIV, HL and pregnancy. One patient received 3 cycles of chemotherapy, refused further treatment, delivered an HIV seropositive girl, and died shortly after from complications of presumed pneumocystis jiroveci pneumonia. The second patient received both active antiretroviral therapy and chemotherapy, delivered an HIV-seronegative boy, and remained in complete remission at 9 months follow-up. We conclude by offering recommendations for the staging and treatment of pregnant, HIV-infected patients with HL. PMID- 20019584 TI - Treatment of castration-resistant prostate cancer: updates on therapeutics targeting the androgen receptor signaling pathway. AB - Androgens play a critical role in the progression of castration-resistant prostate cancer through androgen receptor (AR)-regulated signaling pathways. Progress has been made in the development of potent agents designed to suppress androgen function by blocking the AR, inhibiting the synthesis of androgens, or targeting downstream AR signaling pathways. This review summarizes the development of novel therapies based on current insights into AR signaling pathways in castration-resistant prostate cancer. PMID- 20019585 TI - Association of statins and diabetes mellitus. AB - For years, statins have been used to lower elevated total and low-density lipoprotein cholesterol levels while raising high-density lipoprotein cholesterol levels. Statins work by inhibiting HMG-CoA (3-hydroxy-3-methylglutaryl coenzyme A) reductase, thus stopping the conversion of HMG-CoA to mevalonate, which is found in the cholesterol synthesis cascade. Unfortunately, several studies have shown increased diagnosis and lack of glycemic control of diabetes mellitus when patients are taking high and/or long-term doses of statins, and several mechanisms have been identified that may help explain this side effect. PMID- 20019586 TI - New therapeutic option with N-acetylcysteine for primary sclerosing cholangitis: two case reports. AB - Primary sclerosing cholangitis is a progressive, cholestatic hepatic disease of unknown etiology. It is characterized by progressive inflammation, destruction, and fibrosis of the intrahepatic and extrahepatic bile ducts. Several medical therapies have been tried such as penicilamin, colchicine, methatraxate, cyclosporine, tacrolimus, and ursodeoxycholic acid. Treatment with mucolytic agents in excessively high viscosity conditions appears to have an important role. N-acetylcysteine (NAC), as a mucolytic agent, may fascilitate the drainage in partial obstructions by decreasing the mucous viscosity. We suggest that NAC and ursodeoxycholic acid have markedly positive effects on the clinical course of cholangitis and cholestasis when used together by affecting bile viscosity. Here, we present two cases treated with NAC. NAC capsul therapies at 800 mg/day were administered to two patients with primary sclerosing cholangitis. Clinical and laboratory parameters of patients saw significant improvement. PMID- 20019587 TI - New-onset acute interstitial lung disease after treatment with erlotinib in a patient with metastatic squamous cell carcinoma of the lung. AB - Erlotinib is a Human Epidermal Growth Factor Receptor Type 1/tyrosine kinase (EGFR) inhibitor, which is used for non-small-cell lung cancer treatment. Erlotinib usually has a favorable safety profile however; adverse events such as interstitial lung disease (ILD) have been reported in pivotal studies. ILD usually occurs weeks to months after initiating therapy with Erlotinib. We report a case of Erlotinib induced ILD presenting within 5 days of initiating treatment with Erlotinib. PMID- 20019588 TI - Types of pancreatic cancer in EUS-FNA and chemotherapy. AB - To determine the variety of chemotherapy drugs administrable for malignant pancreatic neoplasm as a result of typification with endoscopic ultrasonography fine needle aspiration (EUS-FNA). A retrospective assessment, in one center, over a period of 1 year. Only malignant pancreatic neoplasm diagnosed by EUS-FNA was recorded. Benign (serous cystic neoplasm) and potentially malignant lesions (mucinous cystic neoplasm and intraductal papillary-mucinous neoplasm) were excluded. Medical data were recorded and Oncological Pharmacy records were studied. Ductal adenocarcinoma were detected in 17 patients (N = 17/22), 2 of them with adenocarcinoma in signet ring and 1 with mucinous adenocarcinoma. The primary therapies used were as follows: Whipple pancreaticoduodenectomy (3), biliary stent by endoscopic retrograde cholangiopancreatography (3), radiological transhepatic percutaneous stent (2), intestinal bypass (2), and a gastric stent (1). The adjuvant drugs used were gemcitabine (10), erlotinib (3), and cetuximab (1), and also radiotherapy was used (1). An unresectable squamous cell carcinoma (N = 1) of the tail was detected, and gemcitabine + vinorelbine + fluorouracil + cisplatin used. Nonfunctioning neuroendocrine tumors were seen in 3 (N = 3) cases and long-acting somatostatin analogues were used (1); the remaining 2 patients showed resectable tumors and were resected accordingly. A metastasis to the pancreatic head in a hepatocellular carcinoma was found in 1 patient (N = 1), allowing specific treatment with sorafenib. Histopathologic analysis with EUS-FNA implies a variety of different treatments. Optimal management was achieved as a result of improved diagnosis, with the advent of new molecular genetic diagnostic methods facilitating the design of specific new therapy and neoadjuvant targeting strategies. PMID- 20019589 TI - Clinical bioequivalence of a dose of clopidogrel Leti Cravid tablets 75 mg versus clopidogrel Sanofi Plavix tablets 75 mg administered on a daily dose for 7 days on healthy volunteers: a clinical trial. AB - Patients undergoing percutaneous coronary intervention procedures, as in patients with coronary disease, should receive treatment indefinitely with acetylsalicylic acid and clopidogrel. New brands of clopidogrel have been developed at lower costs, for helping to avoid premature suspension of antiplatelet therapy, as Cravid Leti Laboratories clopidogrel. Its effectiveness and safety must be compared with Plavix international standard. A prospective, comparative, cross over, and randomized study was conducted in healthy volunteers. Each group received 1 tablet of Clopidogrel Leti or Clopidogrel Sanofi, 75 mg in a single dose daily for 7 days, followed by 7-day washout period before administration of second treatment. Platelet aggregation was measured at the start of each period and at 7 days of treatment through optical aggregometry, using an optical aggregometer 490-2D Chrono-Log, with a self-calibration system working with platelet-rich plasma with readings 0%-100% of light transmission. An important decrease of platelet aggregation was observed in both groups at 7 days of treatment of more than 50%, independent of adenosine diphosphate reactive (Helena and Chrono-Log) used for aggregation (P < 0.05). The relationship between the mean and 90% confidence interval ratio obtained with the 2 different adenosine diphosphate brands were between 80% and 125%, therefore, it can be considered that both brands are bioequivalent and perfectly exchangeable. PMID- 20019590 TI - An elevated level of physical activity is associated with normal lipoprotein(a) levels in individuals from Maracaibo, Venezuela. AB - Coronary artery disease is the main cause of death worldwide. Lipoprotein(a) [Lp(a)], is an independent risk factor for coronary artery disease in which concentrations are genetically regulated. Contradictory results have been published about physical activity influence on Lp(a) concentration. This research aimed to determine associations between different physical activity levels and Lp(a) concentration. A descriptive and cross-sectional study was made in 1340 randomly selected subjects (males = 598; females = 712) to whom a complete clinical history, the International Physical Activity Questionnaire, and Lp(a) level determination were made. Statistical analysis was carried out to assess qualitative variables relationship by chi2 and differences between means by one way analysis of variance considering a P value <0.05 as statistically significant. Results are shown as absolute frequencies, percentages, and mean +/- standard deviation according to case. Physical activity levels were ordinal classified as follows: low activity with 24.3% (n = 318), moderate activity with 35.0% (n = 458), and high physical activity with 40.8% (n = 534). Lp(a) concentration in the studied sample was 26.28 +/- 12.64 (IC: 25.59-26.96) mg/dL. Lp(a) concentration according to low, moderate, and high physical activity levels were 29.22 +/- 13.74, 26.27 +/- 12.91, and 24.53 +/- 11.35 mg/dL, respectively, observing statistically significant differences between low and moderate level (P = 0.004) and low and high level (P < 0.001). A strong association (chi2 = 9.771; P = 0.002) was observed among a high physical activity level and a normal concentration of Lp(a) (less than 30 mg/dL). A lifestyle characterized by high physical activity is associated with normal Lp(a) levels. PMID- 20019591 TI - Adverse effects of multidrug-resistant tuberculosis treatment with a standardized regimen: a report from Iran. AB - Compared with the treatment of drug-sensitive tuberculosis, the treatment of multidrug-resistant tuberculosis (MDR-TB) is more difficult. This study was conducted at the national referral center of tuberculosis in Tehran, Iran, to evaluate adverse drug reactions of treatment of MDR-TB. From 2006 to 2009, all patients admitted into Masih Daneshvari Hospital in Tehran, Iran, for MDR-TB were considered for this study. The standard treatment for MDR-TB consisted of amikacin, prothionamide, ofloxacin, and cycloserine. Ethambutol and pyrazinamide were added to treatment if mycobacterium was sensitive to them. All adverse effects observed in patients were recorded in our registry. Eighty patients were considered in the study; of this cohort, 44 were male and 36 were female. The mean age of patients was 40.64 +/- 17.53 years (range, 14-81 years). All patients received standardized therapy for MDR-TB. The major adverse effects included neurologic side effects (depression, convulsions, consciousness, psychosis, suicide; 7.5%), hepatitis (5%), rash (1.3%), renal toxicity (3.8%), and auditory toxicity (14.5%). Those with neurologic side effects had less favorable outcome (P value = 0.038) and risk of death was increased among them (odds ratio, 13.8; 95% confidence interval, 2.2-86.77). Other adverse effects did not show statistical significance in our analysis. A major adverse effect such as neurologic side effects (depression, convulsions, consciousness, and psychosis) can result in an increased chance of death among patients with MDR-TB. PMID- 20019592 TI - Cognitive function and antihypertensive treatment in the elderly: a 6-year follow up study. AB - Both antihypertensive treatment and statins were proved to reduce mortality and morbidity from cardiovascular disease in the elderly. Yet their effect on cognitive functions of the elderly is unclear. In this study, 518 elderly were interviewed at their home six years ago. BP was measured and antihypertensive drug treatment and cognitive functions were evaluated. 318 of the original sample were re-examined (81% of the survivors). We evaluated the selective survival in relation to changes in BP, the specific drug usage and cognitive functioning. Beta blocker treatment increased the odds of survival but also the odds of decline in MMSE and memory scores. ACE-I treatment was also associated with decline in memory. Subjects treated with CCB had improved MMSE scores, memory, and performed better on tasks requiring concentration. Statins treatment in this study did not show any affect on cognitive functions. Although most subjects were treated by more than one antihypertensive drug, less than half were normalized at baseline and only one quarter at follow-up. Despite the low normalization rate, antihypertensive treatment regimen had been hardly changed during the six years follow-up. The use of statins, however, increased dramatically: from 6% at baseline to almost half at follow-up. According to our findings CCB emerges as the best option for reducing BP in elderly patients. It appeared to improve cognitive functions without hampering survival. PMID- 20019593 TI - Association between apoliprotein E gene polymorphism and hypercholesterolemic phenotype in Maracaibo, Zulia state, Venezuela. AB - Apoliprotein (Apo) E gene polymorphisms have been associated with high plasma lipids levels and cardiovascular disease. The aim of this study was to determine allelic and genotypic frequencies and to evaluate the associations of polymorphisms with hypercholesterolemic phenotypes in a patient population in Maracaibo, Zulia State. Two hundred and twenty-one patients with ages between 9 and 78 years old attending the Endocrine-Metabolic Center at the University of Zulia, Zulia, Venezuela, were recruited. The lipid profile was determined by enzymatic methods. ApoE polymorphisms were determined by polymerase chain reaction-restriction fragment length polymorphism. One hundred and thirty-three dyslipidemic and 88 patients with normal lipids profile were evaluated. The higher proportion of patients corresponded to hypercholesterolemia isolated (46.61%), followed by hypercholesterolemia combined with hypertriglyceridemia and low levels of high-density lipoprotein (21.8%). ApoE epsilon3 allele was the most frequent in the evaluated population (0.80), both in the control group (0.78) and in the dyslipidemic group (0.82), followed by the epsilon4 allele (0.12) for both groups and the epsilon2 allele with values of 0.10 and 0.06, for control and dyslipidemic group, respectively. The epsilon3epsilon3 and epsilon3epsilon4 genotypes were the most frequent in the population, with values of 62.89% and 22.17%, respectively. The genotype frequencies were 57.95% and 66.17% for epsilon3epsilon3; 23.86% and 21.05% for epsilon3epsilon4 in nondyslipidemicos and dyslipidemic patient groups, respectively. The epsilon4epsilon4 genotype was observed only in hypercholesterolemic patients. The homozygote epsilon2epsilon2 and heterozygote epsilon2epsilon3 genotypes were more frequent at the normal lipids profile group, consistent with diverse reports that indicate the association of the epsilon4 allele with elevated cholesterol levels and low cholesterol levels when the epsilon2 allele is present. ApoE polymorphism seems to be associated with variance in serum lipids levels in the population evaluated. PMID- 20019594 TI - Novel mutations identification in exon 4 of LDLR gene in patients with moderate hypercholesterolemia in a Venezuelan population. AB - Familial hypercholesterolemia (FH) is an autosomal dominant disease characterized by increase in low-density lipoprotein (LDL) cholesterol levels and premature coronary artery disease. In Venezuela, the molecular basis of FH has not been characterized, thus, the aim of this study was to investigate mutations in the exon 4 of the LDLR (LDL-receptor) gene in 225 Venezuelan mixed race individuals (65 hypercholesterolemic and 160 normolipidemic). The exon 4 of the LDLR gene was screened by polymerase chain reaction-single strand conformational polymorphism (PCR-SSCP) analysis and DNA sequencing. Additionally, ApoB-100 gene mutations were investigated. Different LDLR gene mutations were identified in 5 hypercholesterolemic patients (7.7%), 3 missense mutations (4.6%), and 2 frameshift mutations (3%). All mutations were heterozygous. The missense mutations included the amino acid substitution p.E180K, p.R194S, and p.C152G. The frameshift mutations are caused by insertions resulting in the creation of stop codons: p.D157fsX158 and p.S173fsX174, which could code for truncated LDLR of 157 and 173 amino acids, respectively. The apoB gene mutations were not detected in any of our patients and to our knowledge 4 mutations identified in this study have not been reported previously, this study being the first comprehensive mutation analysis of the LDLR causing FH in our region. The early identification of individuals at risk allows changes in lifestyle, including dietary intervention, followed by drug treatment. PMID- 20019595 TI - Numerical study of a centrifugal blood pump with different impeller profiles. AB - Computational fluid dynamic simulations of the Kyoto-NTN magnetically suspended centrifugal blood pump with 16 forward-bending blades (16FB), 16 straight blades (16SB), and eight backward-bending blades (8BB) impellers were performed in this study. Commercial CFD software package FLUENT were used as the solver. The purpose of this study is to find out how the impeller blade profiles affect the inner flow and the performance of the centrifugal blood pump. The simulations were carried out with the same impeller rotating speed of 2,000 rpm and pump flow rate of 5 L/min to compare the three pump models. It was found that the 16SB impeller can produce higher pressure head than the 16FB and 8BB impellers under the same impeller rotating speed and pump flow rate. The flow particle tracing was carried out to estimate the blood damage level caused by the three different impeller profiles. It was found that the 16FB and 8BB models have caused the highest and lowest blood damage, respectively. The 16SB is recommended among the three pumps because it can generate the highest pressure head and induce mild blood damage index, although it was higher than that of the 8BB model. PMID- 20019596 TI - Application of drag-reducing polymer solutions as test fluids for in vitro evaluation of potential blood damage in blood pumps. AB - In vitro evaluation of the potential of a circulatory-assist device to damage blood cells has generally been performed using blood from various species. Problems with this approach include the variability of blood sensitivity to mechanical stress in different species, preparation of blood including the adjustment of hematocrit to a standard value, changes in the mechanical properties of blood that occur during storage, and necessity to pool blood samples to obtain an adequate amount of blood for in vitro circulating systems. We investigated whether the mechanical degradation of a drag-reducing polymer (DRP) solution resulting in the loss of drag-reducing ability can indicate the degree of shear-induced blood damage within blood pumps. DRP solution (polyethylene oxide, 4,500 kDa, 1,000 ppm) or porcine blood were driven through a turbulent flow system by a centrifugal pump, either the Bio-Pump BPX-80 (Medtronic, Inc.) or CentriMag (Levitronix LLC) at a constant pressure gradient of 300 mm Hg for 120 minutes. DRP mechanical degradation was evaluated by reduction of flow rate and solution viscosity. A proposed index of DRP mechanical degradation (PDI) is similar to the normalized index of hemolysis (NIH) typically used to quantify the results of in vitro testing of blood pumps. Results indicate that the mechanical degradation of DRP solutions may provide a sensitive standard method for the evaluation of potential blood trauma produced by blood pumps without the use of blood. PMID- 20019597 TI - Noninvasive predictor of HeartMate XVE pump failure by neural network and waveform analysis. AB - Patients increasingly require longer durations of left ventricular assist device (LVAD) therapy. Despite a recent trend toward continuous flow VADs, the HeartMate XVE is still commonly used, but its longevity remains a significant limitation. Existing surveillance methods of pump failure often give inconclusive results. XVE electrical current waveforms were collected regularly (2001-2008) and sorted into quartiles according to number of days until pump failure (Q1, 0-34; Q2, 34 160; Q3, 160-300; and Q4, 300-390 days). Thoratec waveform files were converted into text files. The 10-second electrical current, voltage waveform was identified and isolated for analysis. Waveforms were analyzed by principal component analysis (PCA) and with a fast Fourier transform. Quartiles were compared with analysis of variance (ANOVA). Waveforms (n = 454) were collected for 21 patients with failed pumps. An artificial neural network was used to predict pump failure within 30 days from the waveform characteristics identified though signal processing. PMID- 20019598 TI - Natural anticoagulants can be useful predictors of severity in chronic liver disease. AB - Protein S (PS), protein C (PC), and antithrombin (AT) are produced by the liver, and their levels were previously shown to be reduced in chronic as well as acute liver disease. The aim of this study was to determine whether measurement of PS, PC, and AT levels in patients would be as good as the commonly used clinical and histological parameters of liver disease in discriminating early and advanced hepatocyte dysfunction. A total of 154 patients were recruited and categorized into five groups: hepatitis B inactive carriers in group 1 (n = 29), nonalcoholic steatohepatitis patients in group 2 (n = 30), chronic hepatitis B patients with elevated liver enzymes in group 3 (n = 29), chronic hepatitis C patients with elevated liver enzymes in group 4 (n = 30), liver cirrhosis patients in group 5 (n = 36). There were significant differences between groups in the levels of PC (P = 0.0001), total PS (P = 0.0001), and free PS (P = 0.0001) and AT (P = 0.0001). These parameters were least affected in the control group, then groups 1 and 2, followed by groups 3 and 4, and most affected in group 5. No differences in these tests were detected between groups 1 and 2 and between groups 3 and 4. Univariate and multivariate analyses showed that free PS was the only predictor of significant inflammation (P = 0.0001), and AT (P = 0.001) and PC (P = 0.003) were the most important factors associated with advanced fibrosis. Both PS and PC are sensitive markers of liver disease, but PS is a sensitive marker of liver inflammation, whereas PC may be a more sensitive marker for liver fibrosis. PMID- 20019599 TI - Decreased plasmin resistance by clots of a homophenotypic Aalpha R 16H fibrinogen (Kingsport, slower fibrinopeptide A than fibrinopeptide B release). AB - Reported evidence of a role in fibrinolysis by fibrinopeptide (Fp)B-dependent intermolecular fibrin polymerization contacts and of reversed FpA/FpB release sequence from fibrinogen Kingsport led us to investigate the fibrinolytic properties of Kingsport clots. Clot lysis was induced by either plasmin (pH 7.4) or by a mixture of plasminogen and recombinant tissue plasminogen activator and measured by lysis time and by turbidity (350 nm) time course. Clots were formed by thrombin from plasminogen-free fibrinogen (pH 7.4, 8 mmol/l CaCl2), with or without 40 nmol/l factor XIII or 20% afibrinogenemic plasma. Displaying no differences from corresponding normal controls were (a) lysis of repolymerized fibrin clots, and (b) chromogenic measurements of fibrin-stimulated Glu plasminogen activation by recombinant tissue plasminogen activator. By contrast, thrombin-induced fine and coarse network clots (n = 7) displayed faster turbidity loss than corresponding normal controls and shorter lysis times ranging 31-55% of controls. Comparison of clots of fibrinogen fractions lacking approximately 90% of their alpha chain carboxyl terminal regions, n = 2, also displayed faster plasmin-induced lysis than corresponding controls. To assess the role of FpB release-dependent intermolecular polymerization contacts, clots were prepared in the presence of three molar excess antibeta 15-42 immunoglobulin G, n = 2, and displayed no differences in plasmin-induced lysis from nonimmune immunoglobulin G controls. The reversed FpA/FpB release sequence from Kingsport fibrinogen resulted in clots with decreased resistance to plasmin. We suggest that both markedly slow polymerization and decreased plasmin resistance played causative roles in the hemorrhagic diathesis associated with this dysfibrinogen. PMID- 20019600 TI - Respiratory failure and hypoxemia in the cirrhotic patient including hepatopulmonary syndrome. AB - PURPOSE OF REVIEW: Liver cirrhosis and portal hypertension present with three unique pulmonary complications that are the subject of ongoing clinical research: hepatopulmonary syndrome, portopulmonary hypertension (POPH), and hepatic hydrothorax. The present article is based on a review of the current literature on how to manage these disorders, which are highly important to both anesthesiologists and intensive care physicians. RECENT FINDINGS: Hepatopulmonary syndrome leads to progressive hypoxemia through diffuse vasodilatation of the pulmonary microcirculation. Liver transplantation, although associated with increased mortality, is the only viable treatment. POPH occurs when vascular remodeling triggers an increase in pulmonary artery pressure and resistance. The role of liver transplantation in POPH is controversial given the excessive mortality in patients with moderate to severe POPH. Medical treatment is able to decrease pulmonary artery pressures, though multicenter randomized controlled trials showing improved outcome are lacking to date. Ultrasound plays an increasingly important role in the diagnosis of all three conditions. SUMMARY: Patients with end-stage liver disease are at risk for respiratory failure and hypoxemia and need to be screened for hepatopulmonary syndrome, POPH, and hepatic hydrothorax. Failure to timely recognize and adequately treat these complications of cirrhosis may have severe consequences. PMID- 20019601 TI - Future opportunities and challenges in academic anesthesia in the United Kingdom: a model for maintaining the scientific edge. AB - PURPOSE OF REVIEW: This review outlines the methodology of a major report into academic strategy recently undertaken by the Royal College of Anaesthetists in the United Kingdom. Analyzing the factors that made the report's conclusions robust and workable provides lessons for other countries or healthcare systems faced with similar problems in academic anesthesia. RECENT FINDINGS: The main themes covered by this review include: organization of healthcare and university systems, medical postgraduate training, and funding of research. In the UK, there exists a dual process: clinical service delivery and postgraduate clinical training [which are both the province of the National Health Service (NHS)], and research and undergraduate education (which are both the province of universities). Both NHS and universities are sponsored by the UK government. 'Professors' or 'heads of academic department' in the UK have no automatic responsibility over the activities of the majority of anesthetic consultants in their own hospitals. Anesthesia in the UK has never been a specific unit of assessment in the research granting process, such that anesthetic departments have received little external grant support compared with many other departments such as medicine or cardiology. As a result, anesthetic departments across the UK have either become entrenched as small departments or they have vanished through mergers. SUMMARY: The review's main conclusions are: the creation of a central National Institute for Academic Anaesthesia to coordinate and implement academic strategy and funding; engaging with national pathways for the training of future academics; and suggestions for the future role for anesthetic specialist societies in academic strategy. These initiatives can radically transform the research environment in a positive way. PMID- 20019602 TI - Noninvasive respiratory support in the perioperative period. AB - PURPOSE OF REVIEW: The aims of this review are: to discuss the physiological rationale for noninvasive respiratory support (NRS) in the perioperative period; to give some practical recommendations to safely apply NRS; and to review available clinical data on preventive and curative NRS after cardiac, thoracic and abdominal surgery. RECENT FINDINGS: The most important morbid postoperative pulmonary complication is atelectasis formation, which increases significantly the risk for pneumonia and hypoxic acute respiratory failure. NRS refers to techniques allowing respiratory support without the need for an invasive airway. Two types of NRS are commonly used: noninvasive continuous positive airway pressure (nCPAP) and noninvasive positive pressure ventilation (nPPV). NRS may be an important tool to prevent (prophylactic treatment) or to treat acute respiratory failure avoiding intubation (curative treatment). The aims of NRS are: to partially compensate for the affected respiratory function by reducing the work of breathing; to improve alveolar recruitment with better gas exchange (oxygenation and ventilation); to reduce left ventricular afterload, increasing cardiac output and improving hemodynamics. Evidence suggests that NRS has been proven to be an effective strategy to reduce intubation rates, nosocomial infections, ICU and hospital lengths of stay, morbidity and mortality in postoperative patients. However, before initiating NRS any surgical complications must be treated. SUMMARY: The application of postoperative NRS by a trained and experienced ICU team, with careful patient selection, should optimize patient outcome. PMID- 20019603 TI - Pulmonary hypertension in thoracic surgical patients. AB - PURPOSE OF REVIEW: Literature about thoracic surgery in patients with pulmonary hypertension is scarce. Perceived high risk has appropriately discouraged any unnecessary operation. However, the medical therapy for pulmonary hypertension has made great advances during the last decade. It is likely that future advances in survival and possibly the need for diagnostic procedures will increase the anesthesiologist's exposure to such patients. Understanding the unique physiology as well as new therapeutic agents will facilitate safe care for these challenging patients. RECENT FINDINGS: Since 1998, there have been three World Heath Organization symposiums on pulmonary hypertension. The most recent meeting in 2008 at Dana Point included revisions of the classification scheme and updates on new trials and therapies. New drugs have been utilized in cardiac, lung, or liver transplant operations to treat pulmonary hypertension. It is also recognized that one-lung ventilation presents unique problems for the patient with pulmonary hypertension. Inhalation use of the new pulmonary vasodilator drugs represents a new frontier for intraoperative pharmacology. SUMMARY: Here, the various types of pulmonary hypertension, physiologic changes, and new drug therapies are reviewed. Clinical experience with patients with pulmonary hypertension undergoing both nonthoracic and thoracic procedures is also reviewed. By identifying potential problem areas and application of new pharmacology, an approach to the patient with pulmonary hypertension is synthesized. PMID- 20019604 TI - Heart failure in the ethnic minorities. AB - PURPOSE OF REVIEW: Ethnic minority groups constitute increasing proportions of the population in western countries. Heart failure is increasingly prevalent worldwide and is associated with significant morbidity and mortality. The purpose of this review is to discuss the limited data on heart failure in the ethnic minority groups. RECENT FINDINGS: South Asians have more coronary risk factors that may increase the risk for premature coronary heart disease leading to development of heart failure at a younger age. In the Chinese, hypertension remains an important cause of heart failure and recent data suggest that heart failure with preserved systolic function is common. African-Americans have a higher prevalence of heart failure than whites, present with heart failure at younger ages, and heart failure in them is less likely to be due to coronary heart disease. Findings from a randomized controlled trial conducted specifically on African-Americans support the addition of the combination of isosorbide dinitrate and hydralazine to standard medical regimen for black patients with heart failure. Aboriginal people are more likely than nonaboriginal people to have less access to healthcare and to have a higher disease burden for atherosclerosis. Heart failure is more prevalent in aboriginal than in the nonaboriginal counterparts. SUMMARY: There are important differences across ethnic groups in the causes of heart failure and response to treatment. Given the likely increasing frequency of heart failure in these populations and an increasingly multiethnic world, additional studies on heart failure across different ethnic groups are warranted. PMID- 20019605 TI - Current management of pediatric dilated cardiomyopathy. AB - PURPOSE OF REVIEW: American and European guidelines for treatment of adult heart failure have been recently revised. This review will reconcile those guidelines to recent studies and experience in the treatment of pediatric dilated cardiomyopathy. RECENT FINDINGS: Therapy for pediatric dilated cardiomyopathy includes establishing a diagnosis for diagnostic-specific therapies as well as preventive strategies for anthracycline toxicity and muscular dystrophy. Pediatric studies demonstrate safety and efficacy for use of angiotensin converting enzyme inhibition and beta-blockers in dilated cardiomyopathy. Cardiac resynchronization and mitral annuloplasty represent potential nonpharmacologic therapies. Implantable defibrillator therapy may be of less import in children as compared with adults. Ventricular assist devices (VADs) are now available for all ages, which can improve survival and potentially can lead to recovery. SUMMARY: The robust development of new therapies for adult heart failure has been successfully applied to children with dilated cardiomyopathy. Therapies for severe, intractable heart failure have been more widely utilized than therapies for mild-to-moderate heart failure. PMID- 20019606 TI - Are omega-3 fatty acids options for prevention and treatment of cognitive decline and dementia? AB - PURPOSE OF REVIEW: To report recent data on the potential role of omega-3 fatty acids (n-3 FA) found in oily fish, especially docosahexaenoic acid (DHA), to prevent and treat cognitive decline and Alzheimer's disease. RECENT FINDINGS: Observational studies still provide conflicting results, in which the majority indicate beneficial effects on cognition, both when assessed as a continuous variable or as incident dementia, mainly Alzheimer's disease. Experimental studies have demonstrated potentially ameliorating effects of eicosapentaenoic acid (EPA) and DHA on amyloid fragment formation, signal transduction including upregulation of the apolipoprotein receptor SorLA, as well as on angiogenesis. The role of EPA and DHA metabolites on Alzheimer's disease pathology is under investigation. Recently, three randomized intervention studies, with duration up to 6 months have been reported. In contrast to a small study from Taiwan, no positive overall effects were reported from the Swedish OmegAD Study or from a Dutch study, although post hoc analyses indicate that selected individuals with mild forms of Alzheimer's disease or cognitive decline may respond to treatment. SUMMARY: No firm conclusions can be drawn. Based on epidemiological data, fish including oily fish could be advised as part of a balanced diet for public health purpose, although the evidence for better cognition is only fairly consistent. It is unlikely that n-3 FA will emerge as a treatment option in general for improving cognitive function in patients with Alzheimer's disease. n-3 FA, especially DHA, may turn out as an adjuvant therapy in selected cases. Further long-term intervention studies on individuals with mild cognitive reductions are awaited. PMID- 20019607 TI - Considering energy deficit in the intensive care unit. AB - PURPOSE OF REVIEW: A discrepancy has emerged between experts' recommendations on how to feed ICU patients according to their requirements using parenteral nutrition, if enteral nutrition is not reaching the target. This review describes the differences in the recent guidelines issued by the American Society of Parenteral and Enteral Nutrition (ASPEN) and the European Society of Clinical Nutrition and Metabolism (ESPEN) regarding these aspects. RECENT FINDINGS: ASPEN/Society of Critical Care medicine (SCCM) experts hesitate to recommend the administration of parenteral nutrition to nonmalnourished ICU patients receiving some but not an adequate amount of enteral feeding during the first 7-10 days after admission. ESPEN guidelines recommend to compensate the deficit by adding parenteral nutrition after 24-48 h. These recommendations are mainly based on observational studies showing a strong correlation between negative energy balance and morbidity-mortality. SUMMARY: The energy deficit accumulated by underfed ICU patients during the first days of stay may play an important role in ICU and hospital outcomes for long-staying ICU patients. To reach calorie requirements by artificial nutritional support without harming the patient is still a subject of debate. Future studies, some already on their way will clarify this discussion. PMID- 20019608 TI - The effects of vasopressin and its analogues on the liver and its disorders in the critically ill. AB - PURPOSE OF REVIEW: Vasopressin and terlipressin, a long-acting V1a analogue, are increasingly used in intensive care. The main clinical indications are the treatment of patients with septic shock and of patients with cirrhosis, who develop variceal bleeding, the hepatorenal syndrome or both. In this review, we summarize the effects of these drugs on splanchnic hemodynamics and organ function. RECENT FINDINGS: A recent systematic meta-analysis of randomized trials suggests that terlipressin may improve renal function in hepatorenal syndrome and thereby reduce mortality by 34%. Moreover, a recent study reported that association of terlipressin and albumin was more effective than terlipressin alone. In patients with variceal bleeding, the bleeding control is significantly improved by early administration of terlipressin. The place of vasopressin in the treatment of patients with septic shock is still discussed, but compared with norepinephrine, vasopressin showed at least an equal efficacy. SUMMARY: The use of vasopressin and its synthetic analogues has shown beneficial effects in the management of patients with cirrhosis, especially in the context of variceal bleeding, the hepatorenal syndrome or both. In both cases, the use of terlipressin improved survival. Therefore, in these clinical indications, terlipressin is a part of recommendations. The role of vasopressin in patients with septic shock remains to be precisely evaluated. PMID- 20019609 TI - Lung transplantation: is it still an experimental procedure? AB - PURPOSE OF REVIEW: The number of lung transplants performed worldwide is low and early and late results are worse in comparison with other solid organ transplants. The present review will focus on these two aspects analyzing the causes and describing the possible strategies to overcome these limitations. RECENT FINDINGS: The use of grafts from marginal and from nonheart-beating donors may increase the number of lung transplantation (LTx) with good results. Implementation of donor protocol and optimization of donor management have been reported to be effective in increasing the pool of suitable grafts. Ex-vivo reconditioning technique may be also helpful to better evaluate and recondition usually rejected lungs. This may allow a significant increase in the number of lung transplants performed worldwide. Early and late results of LTx are mainly affected by primary graft dysfunction and the onset of obliterative bronchiolitis. Different strategies have been adopted to reduce the incidence of these two complications with controversial results. SUMMARY: LTx maintains some features of experimental procedure especially in terms of number of performed procedures and early and late results. The various strategies to overcome the limited number of available grafts appear effective but not universally applied and accepted. The different treatments of PDG and obliterative bronchiolitis are still disappointing. To date, the onset of PDG and obliterative bronchiolitis after LTx still significantly impacts on outcomes. A better understanding of the underlying mechanisms in the pathogenesis of primary graft dysfunction and obliterative bronchiolitis may provide improved therapeutic strategies. PMID- 20019610 TI - Managing the toxicities of the aromatase inhibitors. AB - PURPOSE OF REVIEW: The aromatase inhibitors are increasingly used as adjuvant therapy in postmenopausal women with hormone receptor positive breast cancer. With additional experience using these agents, unanticipated side effects have become apparent. Women who experience side effects from adjuvant endocrine therapy are the individuals who derive the greatest benefits. Because noncompliance is highest among those who experience side effects, it is important that these symptoms be palliated. RECENT FINDINGS: The symptomatic effects of aromatase inhibitors include: hot flashes, arthralgias, vaginal dryness and dyspareunia. Hot flashes may successfully be treated with either serotonin reuptake inhibitors or gabapentin. Counseling, vaginal moisturizers and lubricants can improve symptoms related to sexual functioning. The mechanism of arthralgias is uncertain and anti-inflammatory agents are seldom effective. Patients who experience severe musculoskeletal discomfort may necessitate switching to another endocrine agent such as tamoxifen. Physicians should be aware of 'silent' side effects. Screening for bone loss and hypercholesterolemia is critical and patients should be treated accordingly. SUMMARY: Patients and physicians should openly discuss the short and long-term side effects of the aromatase inhibitors as many of these symptoms can be managed effectively. By optimizing quality of life on adjuvant endocrine therapy, noncompliance may be minimized. PMID- 20019611 TI - Use of combined radiation and hyperthermia for gynecological cancer. AB - PURPOSE OF REVIEW: We reviewed all literature on the clinical use of combined radiation and hyperthermia for gynecologic malignancies. RECENT FINDINGS: Combined radiation and hyperthermia should be considered an alternative to chemoradiation for patients with locally advanced cervix cancer and be the first treatment of choice for these patients when radiation cannot be combined with chemotherapy. Several randomized trials have shown an improvement by adding hyperthermia to radiation that is comparable to the improvement found with the addition of chemotherapy to radiation. Hyperthermia does not seem to add to treatment-induced toxicity and the results of hyperthermia are consistent even at 12 years follow-up and could be reproduced in a large, unselected group of cervix cancer patients. A novel indication for combined radiotherapy and hyperthermia is vaginal cancer. Recently, a cohort study showed that the addition of hyperthermia to radiation seems to improve overall survival for patients with vaginal cancer International Federation of Gynecology and Obstetrics stage III. SUMMARY: Combined radiation and hyperthermia should be considered for patients with locally advanced cervix cancer (International Federation of Gynecology and Obstetrics stage IIb and upwards) as an alternative to chemoradiation for patients with a contraindication for chemotherapy. For other patients, the optimal treatment combination is the subject of randomized trials. For vaginal cancer, a prospective registration study is currently ongoing. PMID- 20019612 TI - Recent advances in novel radioimmunotherapeutic approaches for allogeneic hematopoietic cell transplantation. AB - PURPOSE OF REVIEW: Allogeneic hematopoietic cell transplantation (HCT) can be a curative treatment for hematologic malignancies and over the last three decades, novel approaches have resulted in significant reductions in morbidity and mortality. Despite current advances, two major limitations remain: patients continue to die from recurrent disease, and rates of nonrelapse mortality are relatively high due to regimen-related organ toxicities. The ability to target therapy with radiolabeled antibodies provides an innovative way to increase the tumoricidal dose of radiation to tumor sites, whereas sparing normal organs, as further dose escalation of chemotherapy and radiotherapy in HCT preparative regimens is not feasible due to dose-limiting toxicities. RECENT FINDINGS: This review discusses the most current allogeneic HCT data using radioimmunotherapy (RIT) and focuses on recent trials involving patients at the highest risk for relapse. The results from these studies have shown that standard-dose radiolabeled antibodies can be safely combined with reduced-intensity preparative regimens with encouraging results in a single institution phase II study. SUMMARY: Optimism remains that these RIT approaches will improve the cure rates of allogeneic HCT for the thousands of patients with leukemias and lymphomas who undergo this procedure each year. PMID- 20019613 TI - Fungal infections in oncology patients: update on epidemiology, prevention, and treatment. AB - PURPOSE OF REVIEW: Fungal infections that occur after hematopoietic stem cell transplant and therapies for hematologic malignancies contribute to poor outcomes. In the recent year, studies have focused on updating epidemiologic data, providing an understanding of immunogenetic risks for infection, and optimizing prevention and treatment strategies; these studies will be reviewed. RECENT FINDINGS: Contributions to our understanding of epidemiology have come from multicenter studies. Results demonstrate variability in the numbers of diagnosed invasive aspergillosis cases in oncology centers, and a persistent, although relatively low rate of invasive candidiasis. Both multiple and single center studies are demonstrating improved outcomes of treating invasive aspergillosis. Several important studies have outlined the importance of immunogenetics in conferring risks for invasive fungal infections. There have been numerous studies recently published that assist with our understanding of preventive strategies, questioning the utility of pretherapy screening computed tomography scans and masks. Several important randomized trials compared alternative strategies to standard 'empirical' therapies. Finally, several studies were performed that outline complexity in administering new azole antifungals, assisting in optimizing outcomes. SUMMARY: Studies published in the most recent year outline several important advances in our understanding of epidemiology, prevention and treatment of invasive fungal infections. PMID- 20019614 TI - Chronic obstructive pulmonary disease exacerbations in emergency departments: predictors of outcome. AB - PURPOSE OF REVIEW: Exacerbations have a major short-term and long-term impact on patients with chronic obstructive pulmonary disease (COPD). Risk stratification of patients presenting to the emergency department for an exacerbation of COPD is of utmost importance to help in deciding patients' orientation and treatment and in improving outcomes. RECENT FINDINGS: Studies on predictors of outcomes of COPD exacerbations are markedly heterogeneous in terms of assessed variables, outcomes of interest and timeframe (short-term vs. long-term outcomes). Age, severity of underlying disease, clinical signs of immediate severity, and comorbidities are among the most frequently identified prognostic factors of in-hospital outcome. In the most severe patients, ICU scores such as the Acute Physiology and Chronic Health Evaluation II (APACHE II) also have a prognostic value. Several biomarkers have also been studied, but their additional value for risk stratification is not clarified. SUMMARY: Scores predicting the risk of poor outcome could prove useful in the management of COPD exacerbations. Some have been suggested but remain to be further validated before their use can be generalized. PMID- 20019615 TI - Long-acting anticholinergic use in chronic obstructive pulmonary disease: efficacy and safety. AB - PURPOSE OF REVIEW: This article reviews findings from recently published randomized controlled trials and systematic reviews to provide an up-to-date assessment of the efficacy and safety of tiotropium, the only currently available long-acting muscarinic antagonist, when used alone or in conjunction with other respiratory medications in the treatment of chronic obstructive pulmonary disease (COPD). RECENT FINDINGS: Results of recent clinical trials support findings from earlier trials in patients with moderate to very severe COPD demonstrating significant benefits of tiotropium compared to placebo, including sustained increases in lung function, reductions in exacerbations and risk of exacerbation related hospitalizations, and improvement in health status. These benefits were particularly noted in the 4-year UPLIFT study that included 5993 COPD patients, including a large percentage with moderate severity. Whereas the cardiovascular safety of tiotropium has been questioned, results of the UPLIFT trial and a recent pooled analysis of data from 30 trials of tiotropium demonstrated that tiotropium is associated with reductions in the risk of all-cause mortality, cardiovascular mortality and cardiovascular events. SUMMARY: Recent findings confirm the benefits of tiotropium in COPD management and provide reassurance regarding its safety. Moreover, the recent UPLIFT trial provides supportive evidence for the efficacy of tiotropium in COPD patients already receiving treatment with long-acting inhaled beta-agonists and inhaled corticosteroids, suggesting advantages of 'triple' therapy in advanced disease. Further, well designed, adequately powered studies should explore the potential advantages and disadvantages of various combinations of currently available long-acting respiratory medications in COPD, particularly in different clinical phenotypes of this heterogeneous disease. PMID- 20019616 TI - Nonmedicinal therapy in the management of ankle arthritis. AB - PURPOSE OF REVIEW: The incidence of ankle osteoarthritis has increased in recent years, in part, secondary to vehicular trauma. This review describes conservative and operative intervention strategies along with current research related to the management of ankle osteoarthritis. RECENT FINDINGS: Self-reported physical function in patients with ankle osteoarthritis is equivalent to or worse than that of patients with endstage kidney disease, congestive heart failure, or cervical-spine pain and radiculopathy. Nonoperative-intervention strategies such as assistive devices, orthoses, and viscosupplements are frequently used in this clinical population. However, limited objective data are available examining outcomes following nonoperative intervention. Ankle fusion serves as a standard surgical treatment for end-stage ankle osteoarthritis. The limitations of ankle fusion include prolonged immobilization, a relatively high risk of nonunion, and adjacent joint arthritis. Increasing evidence supports the safety and efficacy of total-ankle arthroplasty (TAA). Current (third generation) TAA prostheses feature cementless design and ligament preservation with reduced bone resection and improved instrumentation. SUMMARY: Limited objective evidence exists to guide clinical decision-making related to nonoperative choices such as assistive devices, orthoses, and viscosupplements. Outcomes from prospective clinical trials indicate that newer total ankle-arthroplasty designs provide substantial pain relief in patients with end-stage ankle osteoarthritis. PMID- 20019617 TI - Musculoskeletal rehabilitation in the person with scleroderma. AB - PURPOSE OF REVIEW: The purpose of this review is to examine current evidence for the efficacy of rehabilitation techniques for musculoskeletal impairments in persons with scleroderma. RECENT FINDINGS: The current literature on rehabilitation techniques consists of studies evaluating the effectiveness of paraffin wax treatment, hand and face stretching exercises, connective tissue massage and joint manipulation, splints, and aerobic exercise and resistance training. Only four randomized controlled trials were found and except for those studies, the majority of studies involved small sample sizes and no control groups. However, except for splints, these studies show improvement in joint motion, hand function, and cardiopulmonary endurance. SUMMARY: Although the results from these small studies are promising, larger, randomized controlled studies are needed to fully determine the effectiveness of rehabilitation techniques for persons with scleroderma. PMID- 20019618 TI - Opioid hyperalgesia. AB - PURPOSE OF REVIEW: Opioids are invaluable in the treatment of moderate-to-severe pain. Unfortunately, their prolonged use may be associated with the onset of opioid-induced hyperalgesia (OIH). This review focuses on recent clinical studies that support or refute the existence of OIH in patients. RECENT FINDINGS: Whether or not OIH is a clinical reality is an ongoing debate. In recent years, the majority of clinical trials investigating whether chronic-opioid treatment causes paradoxical pain sensations have been conducted in opioid addicts, patients maintained on methadone and human volunteers receiving acute-morphine infusions. That opioid-maintained patients have different nociceptive profiles compared with opioid naive patients has been both raised and rejected. Independent studies have reinforced the opinion that the development of OIH is based on confounders including pain modality tested, route of drug administration and specific opioid in question. SUMMARY: Improvements in paradoxical pain intensity upon discontinuation of opioid therapy suggests that a multidisciplinary method of pain relief is favoured for chronic-pain patients. Quantitative-sensory testing of pain is offered as the most appropriate way of diagnosing hyperalgesia. We can, thus far only reliably validate the existence of OIH development in normal human volunteers receiving acute-morphine infusions. PMID- 20019619 TI - Role of cervical cordotomy and other neurolytic procedures in thoracic cancer pain. AB - PURPOSE OF REVIEW: Evaluation of the efficacy and safety of the percutaneous cervical cordotomy (PCC) in palliative care medicine in patients suffering from thoracic cancer pain; fluoroscopy-guided versus computed tomographic-guided PCC. Evaluation of recent developments in other neurolytic procedures in thoracic pain. RECENT FINDINGS: Technical progress has provided us with much more accurate means of visualizing the spinal cord and its subunits. Not only do these techniques provide more accuracy in placing the lesion, and thereby increasing safety and efficacy of PCC. There is also no need to use oily contrast media that is incompatible with cerebrospinal fluid and nerve tissue. Recent literature concerning intercostal nerve blocks, selective percutaneous rhizotomy, intrathecal or epidural administration of neurolytic agents is limited. Neurolytic procedures may be improved by ultrasonography-guided procedures. SUMMARY: The results of fluoroscopy-guided PCC are satisfactory with case series reporting complete pain reduction in as much as 82-95% of the patients. For CT guided PCC initial success rates were reported between 80.5-92.5% patients. However, the complication rates and long-term effects if measured and/or mentioned, varied. Hypothetically this technique may be more accurate and therefore probably safer than fluoroscopic-guided PCC. PMID- 20019620 TI - Intracardiac entrapped and pulmonary embolism developing during oral contraceptive therapy in a young woman. AB - We report a case of biatrial thrombus entrapped in a patent foramen ovale (PFO) in a 39-year-old woman under oral contraceptive therapy who was admitted to our hospital with multiple pulmonary embolism without paradoxical embolism. We present this case because of the relatively unusual thromboembolic disease. PMID- 20019621 TI - A late presentation of Dandy-Walker malformation and aortic coarctation. AB - The Dandy-Walker malformation is a rare anomaly of the posterior cranial fossa. Concomitant brain or systemic malformations are frequent and can influence the outcome. Associated cardiac congenital defects usually induce a poor prognosis. We report a case of a 58-yearold man with hydrocephalus, in whom Dandy-Walker malformation was diagnosed, for the first time, after the demonstration of aortic coarctation. This association is very rare and only a few cases have been described; moreover, to our knowledge, this is the first description of this incidence in a middle-aged patient. The characteristic of diseases and physiopathologic features are discussed, focusing attention on the rare and late clinical manifestations. PMID- 20019622 TI - Femoral nailing to treat fractures after lengthening for congenital femoral deficiency in young children. AB - Congenital femoral deficiency in children can be treated with femoral lengthening. A common complication is fracture soon after removal of the external fixator, often despite prophylactic hip spica cast application. These fractures present special challenges because the patients have tight soft tissues and sclerotic intramedullary canals. We treated nine such fractures in eight children (average age, 5.4 years). Most were 'spontaneous' events resulting in transverse fracture through regenerate bone or pin sites. All were stabilized with intramedullary Rush pins using special insertion techniques. Union was achieved (average, 6 weeks); no significant complications occurred. We describe details of the surgical technique. PMID- 20019623 TI - Effects of acute hypoxia on metabolic and hormonal responses to resistance exercise. AB - INTRODUCTION: Several recent studies have shown that resistance exercise combined with vascular occlusion effectively causes increases in muscular size and strength. Researchers speculated that the vascular occlusion-induced local hypoxia may contribute to the adaptations via promoting anabolic hormone secretions stimulated by local accumulation of metabolic subproducts. Here, we examined whether acute systemic hypoxia affects metabolic and hormonal responses to resistance exercise. METHODS: Twelve male subjects participated in two experimental trials: 1) resistance exercise while breathing normoxic air [normoxic resistance exercise (NR)] and 2) resistance exercise while breathing 13% oxygen [hypoxic resistance exercise (HR)]. The resistance exercises (bench press and leg press) consisted of 10 repetitions for five sets at 70% of maximum strength with 1-min rest between sets. Blood lactate, serum growth hormone (GH), epinephrine (E), norepinephrine (NE), insulin-like growth factor 1, testosterone, and cortisol concentrations were measured before normoxia and hypoxia exposures, 15 min after the exposures, and at 0, 15, 30, and 60 min after the exercises. RESULTS: Lactate significantly increased after exercises in both trials (P < 0.05). In the HR trial, GH and cortisol significantly increased after the exercise (P < 0.05) but not in the NR trial. The E, NE, insulin-like growth factor 1, and testosterone significantly increased after the exercises in both trials (P < 0.05). The mean values of lactate, GH, E, and NE after exercises were significantly higher in the HR trial than those in the NR trial (P < 0.05). CONCLUSIONS: These findings suggest that resistance exercise in hypoxic condition caused greater accumulation of metabolites and strong anabolic hormone response. PMID- 20019624 TI - Muscle stiffness and rate of torque development during sprint cycling. AB - PURPOSE: Crank torque (CT) application and rate of CT development (RCTD) are important considerations in sprint cycling. The stiffness of the musculotendinous unit is related to the isometric rate of torque development (RTD); however, this relationship has yet to be examined in sprint cycling. METHODS: Maximal isometric torque (MIT) and isometric RTD of the quadriceps were assessed in 21 trained male cyclists (28.7 +/- 9.5 yr, 1.74 +/- 0.08 m, and 67.5 +/- 7.2 kg). Unilateral musculoarticular (MA) stiffness of the quadriceps was quantified using an oscillation test. Further, the participants performed a maximal 6-s sprint to assess peak power output (POpeak), peak CT (CTpeak), peak RCTD (RCTDpeak), and the crank angles associated with CTpeak and RCTDpeak. Participants were ranked on MA stiffness properties and were divided into a relatively stiff group (SG) and a relatively compliant group (CG). RESULTS: The SG displayed a significantly higher MA stiffness than the CG (P < 0.05). Furthermore, the SG reported significantly elevated MIT (27%), RTD (26%), and RCTDpeak (16%) when compared with the CG (P < 0.05), along with trends for increased POpeak (7%) and CTpeak (8%). The angles at CTpeak and RCTDpeak were 7% and 12% lower for the SG, respectively (P < 0.05). MA stiffness was significantly correlated with RCTDpeak, MIT, RTD, and POpeak. CONCLUSIONS: Higher stiffness is related to superior RCTDpeak in trained cyclists during a single sprint. A significant proportion of the variance in RCTDpeak was attributed to MA stiffness (37%), which was of greater magnitude than the relationship between RCTDpeak and MIT. Furthermore, the lower CTpeak angle and RCTDpeak angle may contribute to a more rapid development of CT. Accordingly, MA stiffness seems to be an important consideration for sprint cycling. PMID- 20019625 TI - Effects of ankle power training on movement time in mobility-impaired older women. AB - PURPOSE: Reduced abilities to generate power put older adults at risk in situations that demand rapid movements. Slower movement times are associated with greater risk of falling and of being involved in a motor vehicle crash. The purpose of this study was to determine the effects of power training on foot movement time and, secondarily, on ankle strength and power in mobility-impaired older women. METHODS: Fifty mobility-impaired women (70-88 yr) trained twice per week for 12 wk in one of three groups (weights, elastic bands, or placebo control). All groups performed seated warm-up exercises, followed by either concentric dorsiflexion (DF) and plantarflexion (PF) resistance exercises (weights and bands) performed "as fast as possible" or upper body flexibility exercises (control). Foot reaction/movement time and ankle DF and PF peak torque (30 degrees x s(-1)) and peak power (90 degrees x s(-1)) were measured before and after training. RESULTS: Participants who trained with elastic bands demonstrated improvements in movement time (decreased by 24 ms or 12%, P = 0.003). All groups demonstrated improvements in DF and PF strength and power, which were not statistically different. CONCLUSIONS: High-velocity/low-load (elastic bands) training improved movement time, which may have important implications in circumstances when rapid generation of torque is required (e.g., to avoid a fall or prevent a vehicle crash). Elastic bands are relatively inexpensive and provide a practical form of training that could be considered in programs designed for older adults with mobility limitations. PMID- 20019626 TI - Antioxidant supplementation does not alter endurance training adaptation. AB - BACKGROUND: There is a considerable commercial market, especially within the sports community, claiming the need for antioxidant supplementation. One argument for antioxidant supplementation in sports is that physical exercise is associated with increased reactive oxygen and nitrogen species (RONS) production, which may cause cell damage. However, RONS production may also activate redox-sensitive signaling pathways and transcription factors, which subsequently, may promote training adaptation. PURPOSE: Our aim was to investigate the effects of combined vitamin C and E supplementation to healthy individuals on different measures of exercise performance after endurance training. METHODS: Using a double-blinded placebo-controlled design, moderately trained young men received either oral supplementation with vitamins C and E (n = 11) or placebo (n = 10) before and during 12 wk of supervised, strenuous bicycle exercise training of a frequency of 5 d x wk(-1). Muscle biopsies were obtained before and after training. RESULTS: After the training period, maximal oxygen consumption, maximal power output, and workload at lactate threshold increased markedly (P < 0.01) in both groups. Also, glycogen concentration, citrate synthase, and beta-hydroxyacyl-CoA dehydrogenase activity in the muscle were significantly higher in response to training (P < 0.01) in both groups. However, there were no differences between the two groups concerning any of the physiological and metabolic variables measured. CONCLUSIONS: Our results suggest that administration of vitamins C and E to individuals with no previous vitamin deficiencies has no effect on physical adaptations to strenuous endurance training. PMID- 20019629 TI - Physiological demands of off-road vehicle riding. AB - INTRODUCTION: The purpose of this study was to characterize the physiological demands of recreational off-road vehicle riding under typical riding conditions using habitual recreational off-road vehicle riders (n = 128). METHODS: Comparisons of the physical demands of off-road vehicle riding were made between vehicle types (all-terrain vehicle (ATV) and off-road motorcycle (ORM)) to the demands of common recreational activities. Habitual riders (ATV = 56, ORM = 72) performed strength assessments before and after a representative trail ride (48 +/- 24.2 min), and ambulatory oxygen consumption was measured during one lap (24.2 +/- 11.8 min) of the ride. RESULTS: The mean VO2 requirement (mL x kg(-1) x min(-1)) while riding an off-road vehicle was 12.1 +/- 4.9 for ATV and 21.3 +/- 7.1 for ORM (P = 0.002), which is comparable to the VO2 required of many common recreational activities. Temporal analysis of activity intensity revealed approximately 14% of an ATV ride and 38% of an ORM ride are within the intensity range (940% VO2 reserve) required to achieve changes in aerobic fitness. Riding on a representative course also led to muscular fatigue, particularly in the upper body. CONCLUSIONS: On the basis of the measured metabolic demands, evidence of muscular strength requirements, and the associated caloric expenditures with off-road vehicle riding, this alternative form of activity conforms to the recommended physical activity guidelines and can be effective for achieving beneficial changes in health and fitness. PMID- 20019628 TI - Neighborhood design and perceptions: relationship with active commuting. AB - PURPOSE: Walking to and from school contributes to total physical activity levels. This study investigated whether perceived and actual neighborhood features were associated with walking to or from school among adolescent girls. METHODS: A sample of geographically diverse eighth-grade girls (N = 890) from the Trial of Activity in Adolescent Girls (TAAG) study living within 1.5 miles of their middle school was recruited. Participants completed a self-administered survey on their neighborhood and walking behavior. Geographic information system data were used to assess objective neighborhood features. Nested multivariable logistic regression analyses were conducted to determine the contribution of perceived and objective measures of walking to or from school. RESULTS: Fifty-six percent (n = 500) of the girls walked to or from school for at least 1 d in a week. White (42%) girls walked more frequently than Hispanic (25%) and African American (21%) girls. Girls were nearly twice as likely to walk to or from school if they perceived their neighborhoods as safe and perceived that they had places they liked to walk, controlling for other potential confounders. In addition, girls who lived closer to school, had more active destinations in their neighborhood, and had smaller-sized blocks were more likely to walk to or from school than those who did not. CONCLUSIONS: Safety, land use, and school location issues need to be considered together when designing interventions to increase walking to and from school. PMID- 20019630 TI - Mood profile of an America's Cup team: relationship with muscle damage and injuries. AB - PURPOSE: To describe the mood profile of an America's Cup sailing team during competition and to evaluate the influence of previous injuries occurrence and intensity of physical work on the boat upon mood state. Relationships between mood domains and metabolic markers of muscle damage were also investigated. METHODS: A descriptive study was conducted on an America's Cup yachting race crew comprising 21 male sailors (mean +/- SD; age = 27.6 +/- 8.5 yr, weight = 89.3 +/- 24.9 kg, BMI = 26.5 +/- 6.9 kg x m(-2)). All measurements were collected during the Louis Vuitton Cup 2007 in Valencia, Spain. The POMS test and creatine kinase (CK) serum activity were measured and correlated. Sailors were grouped according to their presence or absence of previous musculoskeletal injuries (MI) and the intensity of physical work related to boat position: high intensity (HI) and low intensity (LI). RESULTS: According to normative data, pre- and postracing POMS scores were constantly high with prominent anger (24.2 +/- 9 before and 24.9 +/- 10.1 after the race) and depression (22.7 +/- 8.9 before and 20.6 +/- 7.3 after the race). The HI group displayed unchanged anger scores but showed significant differences compared with the LI group (z = -2.07, P = 0.038, mu2 = 0.22) at the end of the competition. The occurrence of a previous injury did not correlate with any interference with mood. Only the fatigue domain before racing had a significant negative correlation with CK levels (r = -0.509, P < 0.05). CONCLUSIONS: The emotional profile of this America's Cup yachting crew showed stable mood scores with high values in anger and depression compared with normative data. Mood was dependent on physical work intensity related to boat position but not on injury occurrence. Enzyme markers of muscle damage had no bearing on most POMS domains, except for fatigue before racing. PMID- 20019631 TI - Effects of acute exercise on sensory and executive processing tasks. AB - PURPOSE: The immediate and delayed effects of a single bout of steady-state aerobic exercise on 19 young adults' (mean = 21.1 yr) sensory sensitivity (critical flicker fusion, CFF) and executive function (modified Paced Auditory Serial Addition Task, PASAT) were assessed. METHODS: Tests were performed before exercise, five times during 40 min of ergometer cycling at 90% ventilatory threshold, and three times during a 30-min postexercise period. In a separate control session, each participant performed the same sequence of tests while seated on the ergometer without pedaling. RESULTS: ANOVA were performed separately on CFF and PASAT scores, which compared performance during exercise and nonexercise conditions at nine time points. Planned ANOVA of CFF scores revealed that the participants' sensory discrimination increased during exercise and then quickly returned to baseline levels immediately after exercise. PASAT scores did not change during or after exercise. CONCLUSIONS: Exercise-induced arousal facilitates sensory processes involved in stimulus detection but does not influence the updating component of executive processing. PMID- 20019632 TI - Carbohydrate supplementation and sex differences in fuel selection during exercise. AB - PURPOSE: To compare the effects of a high-CHO diet (80% CHO) and glucose ingestion (2 g x kg(-1)) during exercise (120 min, 57% VO2max) on fuel selection in women taking (W+OC) or not (W-OC) oral contraceptives and in men (six in each group). METHODS: Substrate oxidation was measured using indirect respiratory calorimetry in combination with a tracer technique to compute the oxidation of exogenous (13C-glucose) and endogenous CHO. RESULTS: In the control situation (mixed diet with water ingestion during exercise), the percent contribution to the energy yield (%En) of CHO oxidation was higher in men than in women (62 vs 53 %En). The high-CHO diet and glucose ingestion during exercise separately increased the %En from CHO oxidation in both men (+12%) and women (+24%), and the sex difference observed in the control situation disappeared. However, the increase in the %En from total CHO oxidation observed when glucose was ingested during exercise and when combined with a high-CHO diet was larger in women than in men (+47 vs +17 %En). This was not attributable to a higher %En from exogenous glucose oxidation in women, for which no sex difference was observed (25 and 27 %En in men and women), but was attributable to a smaller decrease in endogenous glucose oxidation. No significant difference in fuel selection was observed between W+OC and W-OC. CONCLUSIONS: The increase in total CHO oxidation after the high-CHO diet was not different between sexes. Glucose ingestion during exercise, separately and combined to the high-CHO diet, had a greater effect in women than in men; this was mostly attributable to the smaller reduction in endogenous CHO oxidation. PMID- 20019633 TI - Exercise and fitness are related to peripheral nervous system function in overweight adults. AB - PURPOSE: This study examined the association between physical activity and fitness and peripheral nervous system (PNS) function in overweight and obese individuals. METHODS: Forty nondiabetic overweight adults (mean +/- SD; age = 44 +/- 11 yr) were recruited for the study. Peroneal motor nerve and radial, sural, and medial plantar sensory nerve conductions were studied. Maximal oxygen uptake was measured in an incremental bicycle ergometer test. Physical activity was assessed by accelerometer and self-reporting. We analyzed the data using multiple stepwise linear regression models adjusted for age, height, and skin temperature. RESULTS: VO2max predicted 17% of peroneal distal compound muscle action potential (CMAP) amplitude variation and 16% of peroneal proximal CMAP amplitude variation. Physical activity index at the age of 30 yr predicted 9% of peroneal motor nerve conduction velocity (NCV), 8% of peroneal F-wave maximum latency, 14% of medial plantar sensory latency, and 10% of medial plantar sensory NCV variation. CONCLUSIONS: Physical activity and fitness are positively associated with PNS function and should be encouraged in overweight people. PMID- 20019634 TI - Fatigue during maximal sprint cycling: unique role of cumulative contraction cycles. AB - Maximal cycling power has been reported to decrease more rapidly when performed with increased pedaling rates. Increasing pedaling rate imposes two constraints on the neuromuscular system: 1) decreased time for muscle excitation and relaxation and 2) increased muscle shortening velocity. Using two crank lengths allows the effects of time and shortening velocity to be evaluated separately. PURPOSES: We conducted this investigation to determine whether the time available for excitation and relaxation or the muscle shortening velocity was mainly responsible for the increased rate of fatigue previously observed with increased pedaling rates and to evaluate the influence of other possible fatiguing constraints. METHODS: Seven trained cyclists performed 30-s maximal isokinetic cycling trials using two crank lengths: 120 and 220 mm. Pedaling rate was optimized for maximum power for each crank length: 135 rpm for the 120-mm cranks (1.7 m x s(-1) pedal speed) and 109 rpm for the 220-mm cranks (2.5 m x s(-1) pedal speed). Power was recorded with an SRM power meter. RESULTS: Crank length did not affect peak power: 999 +/- 276 W for the 120-mm crank versus 1001 +/- 289 W for the 220-mm crank. Fatigue index was greater (58.6% +/- 3.7% vs 52.4% +/- 4.8%, P < 0.01), and total work was less (20.0 +/- 1.8 vs 21.4 +/- 2.0 kJ, P < 0.01) with the higher pedaling rate-shorter crank condition. Regression analyses indicated that the power for the two conditions was most highly related to cumulative work (r2 = 0.94) and to cumulative cycles (r2 = 0.99). CONCLUSIONS: These results support previous findings and confirm that pedaling rate, rather than pedal speed, was the main factor influencing fatigue. Our novel result was that power decreased by a similar increment with each crank revolution for the two conditions, indicating that each maximal muscular contraction induced a similar amount of fatigue. PMID- 20019635 TI - Muscle deoxygenation during sustained and intermittent isometric exercise in hypoxia. AB - PURPOSE: It is reported that the rate of locomotor muscle fatigue development during intermittent isometric exercise in hypoxia is accelerated compared with normoxia. In contrast, when sustained isometric contractions are used, some studies do not show any effect of hypoxia on fatigue development. Increased intramuscular pressure during sustained isometric exercise causes substantial and sustained ischemia, even in normoxia. Therefore, we hypothesized that the difference in muscle deoxygenation between normoxia and hypoxia would be small during sustained exercise compared with intermittent exercise and that this may contribute to the inconsistent findings. METHODS: Subjects performed sustained and intermittent isometric, unilateral, and submaximal knee-extension exercises (60% maximal voluntary contraction to exhaustion) while breathing normoxic (inspired O2 fraction = 0.21) or hypoxic gas mixtures (inspired O2 fraction = 0.10-0.12). Muscle oxygenation (deoxyhemoglobin/myoglobin and tissue oxygenation index) using near-infrared spectroscopy and surface EMG were measured from the left vastus lateralis. RESULTS: During intermittent isometric exercise in hypoxia, increases in deoxyhemoglobin/myoglobin and reductions of tissue oxygenation index were larger (P < 0.05) than those in normoxia. The rate of rise in integrated EMG during intermittent exercise was accelerated (P < 0.05) in hypoxia. In contrast, there were no significant differences in changes in near infrared spectroscopy variables and integrated EMG during sustained isometric exercise between normoxia and hypoxia. CONCLUSIONS: These results suggest that muscle deoxygenation is exaggerated during intermittent isometric exercise in hypoxia compared with normoxia, whereas during sustained isometric exercise, the extent of muscle deoxygenation is the same between normoxia and hypoxia. The different extent of muscle deoxygenation during sustained and intermittent isometric exercise in normoxia and hypoxia could affect muscle fatigability, which results from the varied rate of accumulation of metabolites. PMID- 20019636 TI - Effect of caffeine ingestion on muscular strength and endurance: a meta-analysis. AB - PURPOSE: Our objective was to perform a systematic review and meta-analysis of the research literature assessing the effect of caffeine ingestion on maximal voluntary contraction (MVC) strength and muscular endurance. METHODS: Thirty-four relevant studies between 1939 and 2008 were included in the meta-analyses of caffeine's effects on MVC strength (n = 27 studies) and muscular endurance (n = 23 studies). Effect sizes (ES) were calculated as the standardized mean difference and meta-analyses were completed using a random-effects model. RESULTS: Overall, caffeine ingestion was found to result in a small beneficial effect on MVC strength (overall ES = 0.19, P = 0.0003). However, caffeine appears to improve MVC strength primarily in the knee extensors (i.e., by approximately 7%, ES = 0.37) and not in other muscle groups such as the forearm or the knee flexors. In an attempt to offer a physiological mechanism behind caffeine's ability to improve MVC strength, a meta-analysis was run on ES from nine studies that measured percent muscle activation during MVC in trials comparing caffeine versus placebo; the overall ES (0.67) was highly significant (P = 0.00008) and of moderate to large size, thus implicating an effect of caffeine on the CNS. Caffeine ingestion was also found to exert a small beneficial effect on muscular endurance (overall ES = 0.28, P = 0.00005). However, it appears caffeine improves muscular endurance only when it is assessed using open (i.e., by approximately 18%, ES = 0.37) and not fixed end point tests. CONCLUSIONS: Overall, caffeine ingestion improves MVC strength and muscular endurance. The effect on strength appears exclusively in the knee extensors, and the effect on muscular endurance appears only detectable with open end point tests. PMID- 20019637 TI - Quadriceps muscle energetics during incremental exercise in children and adults. AB - PURPOSE: This study tested the hypothesis that the muscle metabolic responses of 9- to 12-yr-old children and young adults during incremental quadriceps exercise are dependent on age and sex. METHODS: Fifteen boys, 18 girls, 8 men, and 8 women completed a quadriceps step-incremental test to exhaustion inside a magnetic resonance scanner for determination of the muscle metabolic responses using P magnetic resonance spectroscopy. Quadriceps muscle mass was determined using magnetic resonance imaging scans enabling comparison of metabolic data at a normalized power output. RESULTS: The power output and the energetic state at the Pi/PCr and pH intracellular thresholds (IT) were independent of age and sex. The rate of change in Pi/PCr against power output after the ITPi/PCr (S2) was lower in boys (0.158 +/- 0.089) and girls (0.257 +/- 0.110) compared with men (0.401 +/ 0.114, P < 0.001) and women (0.391 +/- 0.133, P = 0.014), respectively, with sex differences present for children only (P = 0.003). Above the ITpH, S2 was more rapid in the men (-0.041 +/- 0.022, P = 0.003) and girls (-0.030 +/- 0.013, P = 0.011) compared with boys (-0.019 +/- 0.007), with no differences between the girls and the women (-0.035 +/- 0.015, P = 0.479). The increase in Pi/PCr at exhaustion was lower in boys (0.85 +/- 0.38) than that in men (1.86 +/- 0.65, P < 0.001) and in girls (1.78 +/- 1.25) than that in women (4.97 +/- 3.52, P = 0.003), with sex differences in both the child (P = 0.005) and the adult groups (P = 0.019). CONCLUSIONS: During moderate-intensity exercise, muscle metabolism appears adult-like in 9- to 12-yr-old children, although both age- and sex related differences in the "anaerobic" energy turnover are present during high intensity exercise. PMID- 20019638 TI - Resistance training predicts 6-yr body composition change in postmenopausal women. AB - PURPOSE: The aim of this study was to examine the association of exercise frequency (ExFreq) and volume (total weight lifted by military press and squats (SQ)) with change in body composition among postmenopausal women participating in a progressive resistance training study. METHODS: Previously, sedentary women (n = 122, age = 56.3 +/- 4.3 yr) were followed for 6 yr. At 6 yr, there were women who had been randomly assigned to resistance training at baseline (n = 65) controls that were permitted to cross over to the exercise program at 1 yr (n = 32) and 25 true controls. Exercisers and crossovers directed to perform eight core exercises for two sets of eight repetitions at 70%-80% of one-repetition maximum, three times weekly, plus progressive weight bearing, stretching, and balance. Body weight and fat were measured at baseline and annually using anthropometry and dual-energy x-ray absorptiometry. RESULTS: Average change in body weight and total body fat were 0.83 +/- 5.39 and 0.64 +/- 4.95 kg at 6 yr, respectively. In multiple linear regression, ExFreq, military press, and SQ were significantly inversely associated with change in body weight (standardized beta coefficient (SBC) = -0.22 to -0.28, P < 0.01), fat (SBC = -0.25 to -0.33, P < 0.01), and trunk fat (SBC = -0.20 to -0.31, P < 0.03) after adjusting for age, years on hormone therapy, change in lean soft tissue, baseline body composition, and baseline habitual exercise. The lowest tertile of SQ (equivalent to 2.5% attendance) demonstrated significant gain in weight, fat, and trunk fat over 6 yr (P < 0.004), whereas the highest tertile SQ (equivalent to 64% attendance) was able to maintain their weight, total, and regional fat. CONCLUSIONS: We conclude that resistance training is a viable long-term method to prevent weight gain and deleterious changes in body composition in postmenopausal women. PMID- 20019639 TI - Breast elevation and compression decrease exercise-induced breast discomfort. AB - PURPOSE: The aim of this study was to investigate whether a sports bra designed to both elevate and compress the breasts could decrease exercise-induced breast discomfort and bra fit discomfort experienced by women with large breasts relative to a standard encapsulation sports bra. METHODS: Breast kinematic data, bra fit comfort, exercise-induced breast discomfort, and bra rankings in terms of preference to wear during running were compared in 20 women with large breasts who ran on a treadmill under three bra conditions: an experimental bra that incorporated both breast compression and elevation, an encapsulation sports bra, and a placebo bra. Subjective data were collected immediately before and after the treadmill running trials. RESULTS: Exercise-induced breast discomfort (P < 0.01) and bra discomfort (P < 0.01) were significantly less for the experimental bra condition relative to the sports bra and placebo bra. This reduction in discomfort was achieved through greater breast elevation (P < 0.01) and compression, with no difference found in vertical breast displacement (P = 0.12) or vertical breast velocity (P = 0.06). CONCLUSIONS: The design features of greater breast elevation and compression provided significantly increased breast and bra comfort compared with a standard encapsulation sports bra during physical activity for women with large breasts. PMID- 20019640 TI - Short recovery augments magnitude of muscle damage in high responders. AB - PURPOSE: To examine serum creatine kinase (CK) activity after resistance exercise bouts with different rest intervals between sets and exercises in high responding (HR) and normal responding (NR) subjects. METHODS: During each resistance exercise bout, three sets with 10-repetition maximum (10RM) loads were completed for the chest press, cable pulldown, biceps curl, triceps extension, leg extension, and prone leg curl. Each bout differed in the length of the rest interval between sets and exercises, specifically either 1 or 3 min. After blood analysis, subjects were separated into NR or HR on the basis of the peak serum CK activity being in the 90th percentile. RESULTS: The volume completed (load x sets x repetitions) was significantly greater for the 3-min bout versus the 1-min bout, with no significant differences between the HR and the NR groups. For the NR group, serum CK was significantly elevated from 24 to 72 h after each bout, with no significant differences between bouts. Conversely, for the HR group, the 1-min bout resulted in serum CK activity levels that were approximately 70% greater than the 3-min bout at the 48- and 72-h time points. CONCLUSIONS: The key finding from the current study was that the HR group experienced significantly greater CK responses when using shorter rest intervals between sets. Conversely, for the NR group, CK responses were not significantly different between bouts. These findings may have implications for resistance exercise prescription in that some individuals might be less tolerant of shorter rest intervals between sets with greater skeletal muscle microtrauma. PMID- 20019641 TI - Effects of handgrip training with venous restriction on brachial artery vasodilation. AB - Previous studies have shown that resistance training with restricted venous blood flow (Kaatsu) results in significant strength gains and muscle hypertrophy. However, few studies have examined the concurrent vascular responses following restrictive venous blood flow training protocols. PURPOSE: The purpose of this study was to examine the effects of 4 wk of handgrip exercise training, with and without venous restriction, on handgrip strength and brachial artery flow mediated dilation (BAFMD). METHODS: Twelve participants (mean +/- SD: age = 22 +/ 1 yr, men = 5, women = 7) completed 4 wk of bilateral handgrip exercise training (duration = 20 min, intensity = 60% of the maximum voluntary contraction, cadence = 15 grips per minute, frequency = three sessions per week). During each session, venous blood flow was restricted in one arm (experimental (EXP) arm) using a pneumatic cuff placed 4 cm proximal to the antecubital fossa and inflated to 80 mm Hg for the duration of each exercise session. The EXP and the control (CON) arms were randomly selected. Handgrip strength was measured using a hydraulic hand dynamometer. Brachial diameters and blood velocity profiles were assessed, using Doppler ultrasonography, before and after 5 min of forearm occlusion (200 mm Hg) before and at the end of the 4-wk exercise. RESULTS: After exercise training, handgrip strength increased 8.32% (P = 0.05) in the CON arm and 16.17% (P = 0.05) in the EXP arm. BAFMD increased 24.19% (P = 0.0001) in the CON arm and decreased 30.36% (P = 0.0001) in the EXP arm. CONCLUSIONS: The data indicate handgrip training combined with venous restriction results in superior strength gains but reduced BAFMD compared with the nonrestricted arm. PMID- 20019642 TI - Descriptive epidemiology of ambulatory activity in rural, black South Africans. AB - PURPOSE: We investigated the distribution of objectively measured ambulation levels and the association of ambulation levels to adiposity levels in a convenience sample of adolescent and adult, rural black South Africans. METHODS: We analyzed 7-d pedometry data, collected over a period of nine consecutive days, in 789 subjects (women, n = 516; men, n = 273). Adiposity measures included body mass index (BMI) and waist circumference (WC). Obesity was defined as BMI > or = 30 kg x m(-2) or WC > or = 102 cm for men and WC > or = 88 cm for women. RESULTS: The average age- and BMI-adjusted 7-d ambulation level was 12,471 steps per day (95% confidence interval (CI) = 12,107-12,834). Ambulation levels differed between sexes (P = 0.0012), and weekday ambulation differed from weekend ambulation (P = 0.0277). Prevalences, age adjusted to the world population, for sedentarism (SED; <5000 steps per day), low active-somewhat active (5000-9999 steps per day), and active-very active (ACT; > or =10,000 steps per day) were 8.0%, 25.5%, and 66.6%, respectively. In contrast, published self-reported national prevalences for physical inactivity, insufficient physical activity, and physically active have been estimated to be 43%-49%, 20%-27%, and 25%-37%, respectively. After adjusting for sex and age, adiposity measures remained significantly associated with steps per day (BMI, r = -0.08; WC, r = -0.12; P < 0.03). Adjusting for sex, age, village, and season, SED increased the risk of obesity by more than twofold compared with ACT (P < 0.05). Achieving <10,000 steps per day compared with ACT was associated with an increased multivariate adjusted obesity risk of 86%-89% (P < 0.001). CONCLUSIONS: Ambulation levels were high for this rural African sample, and prevalences for SED and ACT differed from published self-reported estimates. PMID- 20019643 TI - Hereditary and environmental contributions to emmetropization and myopia. AB - Twenty years of investigation into emmetropization and the development of myopia has led to several recent insights. Accommodation appears to be an important visual signal for emmetropization. Lens thinning during emmetropization and its cessation at the onset of myopia suggest that interruption of lens stretch during growth is an important part of the process of developing myopia. The ciliary muscle may play a greater role in emmetropization and myopia than previously thought. Time spent outdoors, not near work, may be the more important environmental variable in myopia. The effect of time outdoors shows an important interaction with a substantial genetic contribution to the risk of myopia. PMID- 20019644 TI - Mechanisms of photoreceptor death during retinal degeneration. AB - The development of treatments that slow photoreceptor death could profoundly improve patient wellbeing in those with inherited retinal degenerations. Over recent years, it has emerged that extracellular adenosine-tri-phosphate (ATP) regulates the function of photoreceptors in rodents and primates. Moreover, when the retina is exposed to high levels of ATP, rapid death of photoreceptors occurs, which can be blocked by pretreatment with antagonists to P2X receptors. Compounds that inhibit the action of extracellular ATP slow photoreceptor loss in an animal model of inherited retinal degeneration. In this article, I provide an overview of our work in relation to other research in this area and suggest a model by which ATP contributes to photoreceptor death in inherited retinal degenerations. PMID- 20019645 TI - Early clinical outcomes in children enrolled in human immunodeficiency virus infection care and treatment in lesotho. AB - BACKGROUND: Children are largely underrepresented among those accessing treatment of HIV infection in Africa. Reported outcomes of children enrolled in national care and treatment programs are needed to inform the widespread scale-up of pediatric HIV care in resource-limited settings. METHODS: The objective of this article is to report on the early outcomes of a pediatric HIV infection care and treatment program in Lesotho during its first 14 months of operation. Clinical protocols are described, and characteristics and outcomes of the first cohort of children enrolled in care are reported, derived from a retrospective review of medical records. RESULTS: In the program's first 14 months, 1566 children and adolescents aged between 0 and 16 years were evaluated for HIV, with 567 (36%) confirmed to be infected. Of infected patients, 61% presented with advanced or severe symptoms of HIV disease and 65% presented with CD4 profiles consistent with advanced or severe immunodeficiency, based on World Health Organization 2006 guidelines. Two hundred and eighty four children received highly active antiretroviral therapy. The mortality rate was 18.6 deaths per 100 patient years of follow-up. Ninety-nine percent of deaths occurred within 90 days of enrollment. Deceased patients were significantly younger, had higher rates of stunting and wasting, and were more likely to present with low CD4 cell counts. CONCLUSION: Highly active antiretroviral therapy was well tolerated, but the early mortality rate was high despite concurrent management of HIV and comorbidities. Given that hundreds of thousands of children remain without access to HIV care, renewed efforts are needed to reach this underserved population. PMID- 20019646 TI - Treatment of coronary artery disease in hemodialysis patients evaluated for transplant-a registry study. AB - BACKGROUND: We assessed the results of a noninvasive therapeutic strategy on the long-term occurrence of cardiac events and death in a registry of patients with chronic kidney disease (CKD) and coronary artery disease (CAD). METHODS: We analyzed 519 patients with CKD (56+/-9 years, 67% men, 67% whites) on maintenance hemodialysis with clinical or scintigraphic evidence of CAD by using coronary angiography. RESULTS: In 230 (44%) patients, coronary angiography revealed significant CAD (lumen reduction > or =70%). Subjects with significant CAD were kept on medical treatment (MT; n=184) or referred for myocardial revascularization (percutaneous transluminal coronary angioplasty/coronary artery bypass graft-intervention; n=30) according to American College of Cardiology/American Heart Association guidelines. In addition, 16 subjects refused intervention and were also followed-up. Event-free survival for patients on MT at 12, 36, and 60 months was 86%, 71%, and 57%, whereas overall survival was 89%, 71%, and 50% in the same period, respectively. Patients who refused intervention had a significantly worse prognosis compared with those who actually underwent intervention (events: hazard ratio=4.50; % confidence interval=1.48 15.10; death: hazard ratio=3.39; % confidence interval 1.41-8.45). CONCLUSIONS: In patients with CKD and significant CAD, MT promotes adequate long-term event free survival. However, failure to perform a coronary intervention when necessary results in an accentuated increased risk of events and death. PMID- 20019647 TI - Natural killer cell activation in the lung allograft early posttransplantation. AB - BACKGROUND: In addition to their known antiviral and host defense functions, emerging evidence suggests that natural killer (NK) cells may influence allograft outcomes after solid organ transplantation. Although it is accepted that NK cells are activated in the absence of self-major histocompatibility complex (MHC) class I molecules, little is known of how NK cell dynamics change after transplantation of a MHC disparate lung allograft. MATERIALS AND METHODS: To assess this, we characterized longitudinal changes in NK cell frequency and phenotype, using flow cytometry, both in the peripheral blood and lung allograft in 34 patients undergoing lung transplantation. RESULTS: NK cell frequency decreased with time from transplant with mature NK cells being replaced by a population of less differentiated NK cells expressing lower levels of killer cell immunoglobulin like receptors. In contrast to peripheral blood, NK cells within the allograft consisted of a greater proportion of CD56 cells, expressed less killer cell immunoglobulin-like receptors, and demonstrated an activated phenotype. In clinically stable recipients, peripheral blood NK cells were not activated, however, this contrasted markedly with a small subset of patients experiencing acute allograft rejection or cytomegalovirus reactivation, whose NK cells demonstrated a more activated profile. CONCLUSIONS: Our studies suggest that NK cells become activated after MHC-mismatched lung transplantation. PMID- 20019648 TI - Outcomes of transplantation of single pediatric renal allografts equal to or more than 6 cm in length. AB - BACKGROUND: Because of the donor organ shortage, kidneys from smaller pediatric donors are being increasingly used. However, it is unclear whether small pediatric deceased donor kidneys should be used as single grafts or en bloc. We reviewed our outcomes of single kidney transplants from small pediatric donors into adult recipients. METHODS: Kidneys equal to or more than 6 cm in length were transplanted as a single kidney in seven adult recipients weighing less than 80 kg. Creatinine clearance, kidney graft size, and glomerular size were followed up at 1, 3, 6, and 12 months after transplantation. RESULTS: All patients and grafts, with the exception of one patient, are currently alive with functional grafts. Two kidneys were procured after cardiac death of the donors, but no delayed graft function was observed. A total of 57% (four of seven) patients developed BK viremia, and 29% (two of seven) patients developed BK virus nephropathy. The graft size significantly increased during follow-up by ultrasonography (P=0.02). The renal allograft function by calculated creatinine clearance also significantly improved at 40.6+/-6.9 mL/min, 52.7+/-10.2 mL/min, and 66.2+/-9.7 mL/min at 1, 3, and 12 month after transplantation, respectively (P=0.01). The size of glomeruli significantly increased from 122+/-8.4 mum at 1 to 2 months to 169+/-22.5 mum at 3 to 12 months after transplantation (P<0.01). CONCLUSIONS: Kidneys equal to or more than 6 cm from small pediatric donors can be successfully transplanted as a single kidney. Single pediatric kidney transplantation can provide adequate renal function with a speedy increase in allograft size. PMID- 20019650 TI - Use of the levonorgestrel-releasing intrauterine system: an Austrian perspective. AB - Approximately 12 million women worldwide use the levonorgestrel-releasing intrauterine system (IUS), with approximately 180,000 users of this IUS currently reported in Austria. A patient satisfaction study of 591 women in Austria revealed a high number of 'very satisfied' (79%) and 'satisfied' (19%) patients. Reliability, comfort, excellent compatibility and less severe, shorter and less painful monthly periods were the most frequently named advantages of the levonorgestrel-releasing IUS. Medication-induced cervical priming before insertion can be carried out on a routine or selective basis (for example in nullipara, in women who have undergone cervical conisation or in women who have previously experienced painful insertion). There is, at present, no evidence of an increased rate of breast cancer through use of the levonorgestrel-releasing IUS. A directly comparative study with oral contraceptives in young nullipara showed excellent results for the levonorgestrel-releasing IUS, with no perforations, inflammation or pregnancies. PMID- 20019649 TI - Decreased transplant arteriosclerosis in endothelial nitric oxide synthase deficient mice. AB - BACKGROUND: Occlusive vascular changes, characterized by the formation of a neointima with lumen obstruction, are key histologic findings of allograft arteriosclerosis. Vascular integrity of the graft is critically dependent on nitric oxide (NO), synthesized by NO synthases (NOS), of which three isoforms have been located in the arterial wall: endothelial NOS (eNOS), inducible NOS, and neuronal NOS (nNOS). We have studied the role of NOS in a murine model of aortic allograft rejection. METHODS: The descending thoracic aorta of donor mice (BALB/c mice) was transplanted into two groups of recipients: (a) C57BL/6J and (b) C57BL/6J mice homozygous (-/-) for a knockout of the eNOS gene (eNOS(-/-)). RESULTS: After 4 weeks, pronounced neointima formation, upregulated expression of adhesion molecules, and increased infiltration by inflammatory cells were demonstrated in wild-type recipient mice, whereas eNOS(-/-) recipient mice were protected from neointima development by a significantly increased synthesis of NO, as shown by increased formation of cGMP; this was mainly explained by upregulation of inducible NOS and nNOS. CONCLUSIONS: Upregulation of inducible NOS and nNOS isoforms may be beneficial in preventing allograft arteriosclerosis in the early posttransplant period. PMID- 20019653 TI - Assessment of epidemiology capacity in State Health Departments - United States, 2009. AB - Since 2001, the Council of State and Territorial Epidemiologists (CSTE) periodically has conducted a standardized national assessment of state health departments' core epidemiology capacity. During April-June 2009, CSTE sent a web based questionnaire to the state epidemiologist in each of the 50 states and the District of Columbia. The assessment inquired into workforce capacity and technological advancements to support surveillance. Measures of capacity included total number of epidemiologists and self-assessment of the state's ability to carry out four essential services of public health (ESPH). This report summarizes the results of the assessment, which determined that in 2009, 10% fewer epidemiologists were working in state health departments than in 2006. Compared with 2006, the percentage of state health departments with substantial-to-full (>50%) epidemiology capacity decreased in three ESPH, including 1) capacities to monitor and detect health problems, 2) investigate them, and 3) evaluate the effectiveness of population-based services. The percentage of departments with substantial-to-full epidemiology capacity for bioterrorism/emergency response decreased slightly, from 76% in 2006 to 73% in 2009. More than 30% of states reported minimal-to-no (<25%) capacity to evaluate and conduct research and for five of nine epidemiology program areas, including environmental health, injury, occupational health, oral health, and substance abuse. Working together, federal, state, and local agencies should develop a strategy to address downward trends and major gaps in epidemiology capacity. PMID- 20019651 TI - Rational of the use of aliskiren in hypertension and beyond. AB - Arterial hypertension is an independent risk factor for cardiovascular diseases and one of the major causes for mortality worldwide. Drugs, that control hypertension effectively are therefore needed to reduce hypertension induced morbidity and mortality. The inhibition of the renin-angiotensin-aldosterone system (RAAS) is one target to control blood pressure in these patients. The new direct renin inhibitor aliskiren is one new substance on the market to inhibit the RAAS effectively by suppression of the plasma renin activity, which inhibits the RAAS at its rate-limiting step. Therefore, aliskiren in monotherapy and in combination might yield beneficial effects for the patients. Nevertheless, blood pressure lowering has to be combined with a reduction of target organ damage for all drug classes prescribed to patients with hypertension. Therefore, we review here the major effects of this new drug not only in regard to hypertension but also in regard to target organ damage reduction and possible changes in morbidity and mortality, which future trials will investigate. PMID- 20019654 TI - Imported case of Marburg hemorrhagic fever - Colorado, 2008. AB - Marburg hemorrhagic fever (MHF) is a rare, viral hemorrhagic fever (VHF); the causative agent is an RNA virus in the family Filoviridae, and growing evidence demonstrates that fruit bats are the natural reservoir of Marburg virus (MARV). On January 9, 2008, an infectious disease physician notified the Colorado Department of Public Health and Environment (CDPHE) of a case of unexplained febrile illness requiring hospitalization in a woman who had returned from travel in Uganda. Testing of early convalescent serum demonstrated no evidence of infection with agents that cause tropical febrile illnesses, including VHF. Six months later, in July 2008, the patient requested repeat testing after she learned of the death from MHF of a Dutch tourist who had visited the same bat roosting cave as the patient, the Python Cave in Queen Elizabeth National Park, Uganda. The convalescent serologic testing revealed evidence of prior infection with MARV, and MARV RNA was detected in the archived early convalescent serum. A public health investigation did not identify illness consistent with secondary MHF transmission among her contacts, and no serologic evidence of infection was detected among the six tested of her eight tour companions. The patient might have acquired MARV infection through exposure to bat secretions or excretions while visiting the Python Cave. Travelers should be aware of the risk for acquiring MHF in caves or mines inhabited by bats in endemic areas in sub-Saharan Africa. Health-care providers should consider VHF among travelers returning from endemic areas who experience unexplained febrile illness. PMID- 20019655 TI - Agranulocytosis associated with cocaine use - four States, March 2008-November 2009. AB - In April 2008, a clinical reference laboratory in New Mexico notified the New Mexico Department of Health (NMDOH) of a cluster of unexplained agranulocytosis cases confirmed by bone marrow histopathology during the preceding 2 months. NMDOH began an investigation, which identified cocaine use as a common exposure in 11 cases of otherwise unexplained agranulocytosis during April 2008-November 2009. In the midst of the NMDOH investigation, in November 2008, public health officials in British Columbia and Alberta, Canada, reported detecting levamisole (an antihelminthic drug used mainly in veterinary medicine and a known cause of agranulocytosis) from clinical specimens and drug paraphernalia of cocaine users with agranulocytosis. In January 2009, NMDOH posted a notification of its findings on CDC's Epidemic Information Exchange (Epi-X) and notified poison control centers. In a separate investigation during April--November 2009, public health officials in Seattle, Washington, identified 10 cases of agranulocytosis among persons with a history of cocaine use. Of the 21 cases, levamisole was detected from clinical specimens in four of the five patients tested. According to the Drug Enforcement Administration (DEA), as of July 2009, 69% of seized cocaine lots coming into the United States contained levamisole as an added agent. This report summarizes the investigations in New Mexico and Washington, which suggested that levamisole in cocaine was the likely cause of the agranulocytosis. Health-care providers should consider these findings in the differential diagnosis of agranulocytosis, and public health officials should be aware of cases of agranulocytosis associated with cocaine use. PMID- 20019657 TI - Applied research deserves Nobel prizes. PMID- 20019656 TI - PuraMatrix encapsulation of cancer cells. AB - Increasing evidence suggests that culturing cancer cells in three dimensions more accurately recapitulates the complexity of tumor biology. Many of these models utilize reconstituted basement membrane derived from animals which contain a variable amount of growth factors and cytokines that can influence the growth of these cell culture models. Here, we describe in detail the preparation and use of PuraMatrix, a commercially available self assembling peptide gel that is devoid of animal-derived material and pathogens to encapsulate and propagate the ovarian cancer cell line, OVCAR-5. We begin by describing how to prepare the PuraMatrix prior to use. Next, we demonstrate how to properly mix the PuraMatrix and cell suspension to encapsulate the cells in the hydrogel. Upon the addition of cell culture media or injection into a physiological environment, the peptide component of PuraMatrix rapidly self assembles into a 3D hydrogel that exhibits a nanometer scale fibrous structure with an average pore size of 5-200 nm(1). In addition, we demonstrate how to propagate cultures grown in encapsulated PuraMatrix. When encapsulated in PuraMatrix, OVCAR-5 cells assemble into three dimensional acinar structures that more closely resemble the morphology of micrometastatic nodules observed in the clinic than monolayer in vitro models. Using confocal microscopy we illustrate the appearance of representative OVCAR-5 cells encapsulated in PuraMatrix on day 1, 3, 5, and 7 post plating. The use of PuraMatrix to culture cancer cells should improve our understanding of the disease and allow us to assess treatment response in more clinically predictive model systems. PMID- 20019659 TI - Plasmonics: Electrifying plasmonics on silicon. PMID- 20019661 TI - Material witness: Why is boron so hard? PMID- 20019660 TI - Cell rheology: Stressed-out stem cells. PMID- 20019662 TI - Organic crystals: Packing down. PMID- 20019663 TI - Nanotribology: The renaissance of friction. PMID- 20019664 TI - Dissolution of insulating oxide materials at the molecular scale. AB - Our understanding of mineral and glass dissolution has advanced from simple thermodynamic treatments to models that emphasize adsorbate structures. This evolution was driven by the idea that the best understanding is built at the molecular level. Now, it is clear that the molecular questions cannot be answered uniquely with dissolution experiments. At the surface it is unclear which functional groups are present, how they are arranged, and how they interact with each other and with solutes as the key bonds are activated. An alternative approach has developed whereby reactions are studied with nanometre-sized aqueous oxide ions that serve as models for the more complicated oxide interface. For these ions, establishing the structure is not a research problem in itself, and bond ruptures and dissociations can be followed with much confidence. We review the field from bulk-dissolution kinetics to the new isotope-exchange experiments in large oxide ions. PMID- 20019665 TI - Rapid reprogramming of haemoglobin structure-function exposes multiple dual antimicrobial potencies. AB - The intrinsic cytotoxicity of cell-free haemoglobin (Hb) has hampered the development of reliable Hb-based blood substitutes for over seven decades. Notably, recent evidence shows that the Hb deploys this cytotoxic attack against invading microbes, albeit, through an unknown mechanism. Here, we unraveled a rapid molecular reprogramming of the Hb structure-function triggered by virulent haemolytic pathogens that feed on the haem-iron. On direct contact with the microbe, the Hb unveils its latent antimicrobial potency, where multiple antimicrobial fragments are released, each harbouring coordinated 'dual-action centres': microbe binding and pseudoperoxidase (POX) cycle activity. The activated Hb fragments anchor onto the microbe while the juxtaposed POX instantly unleashes a localized oxidative shock, killing the pathogen-in-proximity. This concurrent action conceivably restricts the diffusion of free radicals. Furthermore, the host astutely protects itself from self-cytotoxicity by simultaneously releasing endogenous antioxidants. We found that this decryption mechanism of antimicrobial potency is conserved in the ancient invertebrate respiratory protein, indicating its fundamental significance. Our definition of dual-antimicrobial centres in the Hb provides vital clues for designing a safer Hb-based oxygen carrier blood substitute. PMID- 20019666 TI - DNA polymerase beta is critical for mouse meiotic synapsis. AB - We have shown earlier that DNA polymerase beta (Pol beta) localizes to the synaptonemal complex (SC) during Prophase I of meiosis in mice. Pol beta localizes to synapsed axes during zygonema and pachynema, and it associates with the ends of bivalents during late pachynema and diplonema. To test whether these localization patterns reflect a function for Pol beta in recombination and/or synapsis, we used conditional gene targeting to delete the PolB gene from germ cells. We find that Pol beta-deficient spermatocytes are defective in meiotic chromosome synapsis and undergo apoptosis during Prophase I. We also find that SPO11-dependent gammaH2AX persists on meiotic chromatin, indicating that Pol beta is critical for the repair of SPO11-induced double-strand breaks (DSBs). Pol beta deficient spermatocytes yielded reduced steady-state levels of the SPO11 oligonucleotide complexes that are formed when SPO11 is removed from the ends of DSBs, and cytological experiments revealed that chromosome-associated foci of replication protein A (RPA), RAD51 and DMC1 are less abundant in Pol beta deficient spermatocyte nuclei. Localization of Pol beta to meiotic chromosomes requires the formation of SPO11-dependent DSBs. Taken together, these findings strongly indicate that Pol beta is required at a very early step in the processing of meiotic DSBs, at or before the removal of SPO11 from DSB ends and the generation of the 3' single-stranded tails necessary for subsequent strand exchange. The chromosome synapsis defects and Prophase I apoptosis of Pol beta deficient spermatocytes are likely a direct consequence of these recombination defects. PMID- 20019667 TI - Interactions of PAN's C-termini with archaeal 20S proteasome and implications for the eukaryotic proteasome-ATPase interactions. AB - Protein degradation in the 20S proteasome is regulated in eukaryotes by the 19S ATPase complex and in archaea by the homologous PAN ATPase ring complex. Subunits of these hexameric ATPases contain on their C-termini a conserved hydrophobic tyrosine-X (HbYX) motif that docks into pockets in the 20S to stimulate the opening of a gated substrate entry channel. Here, we report the crystal structure of the archaeal 20S proteasome in complex with the C-terminus of the archaeal proteasome regulatory ATPase, PAN. This structure defines the detailed interactions between the critical C-terminal HbYX motif and the 20S alpha subunits and indicates that the intersubunit pocket in the 20S undergoes an induced-fit conformational change on binding of the HbYX motif. This structure together with related mutagenesis data suggest how in eukaryotes certain proteasomal ATPases bind to specific pockets in an asymmetrical manner to regulate gate opening. PMID- 20019668 TI - mNUDC is required for plus-end-directed transport of cytoplasmic dynein and dynactins by kinesin-1. AB - Lissencephaly is a devastating neurological disorder caused by defective neuronal migration. The LIS1 (or PAFAH1B1) gene was identified as the gene mutated in lissencephaly patients, and was found to regulate cytoplasmic dynein function and localization. In particular, LIS1 is essential for anterograde transport of cytoplasmic dynein as a part of the cytoplasmic dynein-LIS1-microtubule complex in a kinesin-1-dependent manner. However, the underlying mechanism by which a cytoplasmic dynein-LIS1-microtubule complex binds kinesin-1 is unknown. Here, we report that mNUDC (mammalian NUDC) interacts with kinesin-1 and is required for the anterograde transport of a cytoplasmic dynein complex by kinesin-1. mNUDC is also required for anterograde transport of a dynactin-containing complex. Inhibition of mNUDC severely suppressed anterograde transport of distinct cytoplasmic dynein and dynactin complexes, whereas motility of kinesin-1 remained intact. Reconstruction experiments clearly demonstrated that mNUDC mediates the interaction of the dynein or dynactin complex with kinesin-1 and supports their transport by kinesin-1. Our findings have uncovered an essential role of mNUDC for anterograde transport of dynein and dynactin by kinesin-1. PMID- 20019669 TI - Molecular basis for antagonism between PDGF and the TGFbeta family of signalling pathways by control of miR-24 expression. AB - Modulation of the vascular smooth-muscle-cell (vSMC) phenotype from a quiescent 'contractile' phenotype to a proliferative 'synthetic' phenotype has been implicated in vascular injury repair, as well as pathogenesis of vascular proliferative diseases. Both bone morphogenetic protein (BMP) and transforming growth factor-beta (TGFbeta)-signalling pathways promote a contractile phenotype, while the platelet-derived growth factor-BB (PDGF-BB)-signalling pathway promotes a switch to the synthetic phenotype. Here we show that PDGF-BB induces microRNA 24 (miR-24), which in turn leads to downregulation of Tribbles-like protein-3 (Trb3). Repression of Trb3 coincides with reduced expression of Smad proteins and decrease in BMP and TGFbeta signalling, promoting a synthetic phenotype in vSMCs. Inhibition of miR-24 by antisense oligonuclotides abrogates the downregulation of Trb3 as well as pro-synthetic activity of the PDGF-signalling pathway. Thus, this study provides a molecular basis for the antagonism between the PDGF and TGFbeta pathways, and its effect on the control of the vSMC phenotype. PMID- 20019671 TI - Obstructive sleep apnea, masked hypertension, and arterial stiffness in men. AB - BACKGROUND: Obstructive sleep apnea (OSA) is an established cause of hypertension. However, it is not clear whether the frequency of masked hypertension in patients with OSA and whether OSA have an independent role on arterial stiffness taking into account ambulatory blood pressure (BP) monitoring (ABPM). METHODS: We evaluated 61 male normotensive participants as determined by casual clinic BP level <140/90 mm Hg without clinical evidence of cardiovascular disease and on no medications (43 patients with moderate-to-severe OSA (apnea hypopnea index (AHI) > or = 15 events/hour by polysomnography) and 18 age- and body mass index-matched controls without OSA (AHI <5 events/hour)). Pulse wave velocity (PWV), an index of arterial stiffness, and 24-h ABPM were performed in a blinded fashion. Masked hypertension was defined when abnormal daytime ABPM was > or = 135 or > or = 85 mm Hg. RESULTS: The AHI and lowest oxygen saturation were 2.6 +/- 1.6 and 90 +/- 2 vs. 52.8 +/- 21.0 events/hour and 75 +/- 10% for controls and OSA patients, respectively; P < 0.001. Compared with controls, patients with OSA had higher office systolic BP (113 +/- 9 vs. 118 +/- 10 mm Hg; P = 0.05) and a higher unadjusted proportion of masked hypertension (2 controls (11.1%) vs. 13 patients (30.2%); P < 0.05). PWV was 8.7 +/- 0.7, 9.4 +/- 1.0, and 10.6 +/- 1.1 m/s in the control, OSA without and with masked hypertension groups, respectively (P < 0.01 for each comparison). Multiple regression showed that systolic daytime ABPM and the lowest oxygen saturation were independently related to PWV (adjusted R2 = 0.34; P < 0.01). CONCLUSIONS: Patients with OSA presented a higher unadjusted rate of masked hypertension than matched controls. Lowest oxygen saturation has an independent association with arterial stiffness. PMID- 20019670 TI - Aortic PWV in chronic kidney disease: a CRIC ancillary study. AB - BACKGROUND: Aortic pulse wave velocity (PWV) is a measure of arterial stiffness and has proved useful in predicting cardiovascular morbidity and mortality in several populations of patients, including the healthy elderly, hypertensives and those with end-stage renal disease receiving hemodialysis. Little data exist characterizing aortic stiffness in patients with chronic kidney disease (CKD) who are not receiving dialysis, and in particular the effect of reduced kidney function on aortic PWV. METHODS: We performed measurements of aortic PWV in a cross-sectional cohort of participants enrolled in the Chronic Renal Insufficiency Cohort (CRIC) study to determine factors which predict increased aortic PWV in CKD. RESULTS: PWV measurements were obtained in 2,564 participants. The tertiles of aortic PWV (adjusted for waist circumference) were <7.7 m/s, 7.7 10.2 m/s, and >10.2 m/s with an overall mean (+/- s.d.) value of 9.48 +/- 3.03 m/s (95% confidence interval = 9.35-9.61 m/s). Multivariable regression identified significant independent positive associations of age, blood glucose concentrations, race, waist circumference, mean arterial blood pressure, gender, and presence of diabetes with aortic PWV and a significant negative association with the level of kidney function. CONCLUSIONS: The large size of this unique cohort, and the targeted enrollment of CKD participants provides an ideal situation to study the role of reduced kidney function as a determinant of arterial stiffness. Arterial stiffness may be a significant component of the enhanced cardiovascular risk associated with kidney failure. PMID- 20019672 TI - Effects of exenatide on systolic blood pressure in subjects with type 2 diabetes. AB - BACKGROUND: The majority of patients with type 2 diabetes mellitus have blood pressure (BP) exceeding the recommended value of <130/80 mm Hg. Optimal control of hyperglycemia and hypertension has been shown to reduce the incidence of macrovascular and microvascular complications due to diabetes. Treatment with the GLP-1 receptor agonist exenatide, previously demonstrated to reduce hemoglobin A(1C) and weight in subjects with type 2 diabetes, was associated with BP reduction in several studies. METHODS: This analysis explored the effects of exenatide vs. placebo or insulin on BP measurements in pooled data from six trials including 2,171 subjects studied for at least 6 months. RESULTS: Overall, 6 months of exenatide treatment was associated with a significantly greater reduction in systolic BP (SBP) compared with placebo (least squares mean (s.e.): difference of -2.8 mm Hg (0.75); P = 0.0002) or insulin (difference of -3.7 mm Hg (0.85); P < 0.0001). No significant intergroup differences in diastolic BP (DBP) were observed. The majority of the intergroup difference was observed in subjects with SBP > or = 130 mm Hg (difference of -3.8 mm Hg (1.08) from placebo: P = 0.0004; difference of -4.0 mm Hg (1.01) from insulin; P < 0.0001). The largest intertreatment differences between exenatide and comparators were observed in subjects with SBP >/=150 mm Hg. Similar responses were observed in African American subjects. A weak correlation between the amount of weight lost and reduction in SBP was found (r = 0.09, P = 0.002) for exenatide-treated subjects. CONCLUSIONS: These results support the need for a prospective, randomized, controlled study of BP changes during exenatide treatment in patients with hypertension and type 2 diabetes. PMID- 20019674 TI - Role of oxidative stress in erythropoietin-induced hypertension in uremic rats. AB - BACKGROUND: Erythropoietin (EPO) administration in uremic rats leads to an increase in blood pressure (BP). Because chronic renal failure has been associated with oxidative stress, we hypothesize that EPO treatment could accentuate this condition and contribute to hypertension. The present study was designed to investigate the role of reactive oxygen species in EPO-induced hypertension and the effect of tempol, a superoxide dismutase-mimetic. METHODS: Renal failure was induced by a two-stage 5/6 nephrectomy followed by a 3-week stabilization period. Uremic rats were divided into four groups and received for 4 weeks: vehicle; EPO (100 U/kg, subcutaneously, three times per week); vehicle + tempol (1 mmol/l in drinking water); and EPO + tempol. Systolic BP and biochemical parameters were assessed before and at the end of the treatment. Renal histology, creatinine clearance rate, endothelin-1 (ET-1) concentrations and superoxide anion production were assessed at the end of the study. RESULTS: The uremic rats developed anemia and hypertension. ET-1 concentrations and superoxide anion production were increased. EPO administration corrected anemia, but accentuated hypertension and renal injuries such as glomerulosclerosis, interstitial fibrosis, and inflammation. EPO therapy further increased tissue levels of ET-1 and superoxide anion production. Tempol treatment improved hypertension and renal injury, and reduced ET-1 concentrations and superoxide anion production. CONCLUSION: Oxidative stress contributes to the development of hypertension and to the progression of renal injuries in uremic rats. EPO administration further increases oxidative stress, which might partly account for the accentuation of hypertension and renal injury. PMID- 20019673 TI - Leptin, not adiponectin, predicts hypertension in the Copenhagen City Heart Study. AB - BACKGROUND: Leptin and adiponectin are hormones secreted by adipose tissue, and both hormones are candidate intermediaries between adipose tissue and overweight related diseases. So far, no prospective study has been published where the independent effects of these two hormones on the development of hypertension have been directly compared. The objective of this study was to investigate the relationships between plasma levels of leptin and adiponectin and new-onset hypertension in the Copenhagen City Heart Study (CCHS). METHODS: In a prospective study design, we examined new-onset hypertension in 620 women and 300 men who were normotensive in the third CCHS examination, which was performed in 1991 1994. RESULTS: Between the third and the fourth CCHS examination, which was performed in 2001-2003, 254 had developed hypertension, defined as systolic blood pressure (SBP) > or = 140 mm Hg, or diastolic blood pressure (DBP) > or = 90 mm Hg, or use of antihypertensive medication. Using logistic regression analysis, adjusting for age, sex, estimated glomerular filtration rate, triglycerides, high density lipoprotein cholesterol (HDL-C), fibrinogen, and glucose, and with leptin and adiponectin included in the same model, leptin was significantly associated with new-onset hypertension with an odds ratio (95% confidence interval) of 1.28 (1.08-1.53; P < 0.005) for 1 s.d. higher level of log-transformed leptin, whereas adiponectin was not significantly associated with new-onset hypertension having an odds ratio of 1.02 (0.84-1.24; P = 0.83) for 1 s.d. higher level of log transformed adiponectin. CONCLUSIONS: In the CCHS, leptin, but not adiponectin, was a significant independent predictor of new-onset hypertension. PMID- 20019675 TI - Utilizing Ecological Momentary Assessment in pediatric obesity to quantify behavior, emotion, and sleep. AB - This study examined the feasibility of using Ecological Momentary Assessment (EMA) to examine important domains relevant to interregulatory health processes in overweight adolescent females in their natural environments. Participants were 20 overweight adolescent females engaged in a cognitive-behavioral and motivational interviewing intervention aimed at weight loss and improving mood (11-19 years old, 80% white, 15% African American, mean BMI = 39). During this EMA protocol, participants were asked to report their physical activity (PA), nutrition, mood, and sleep during 14 cellular phone calls over three extended weekends (Thursday to Monday). Simultaneously, participants wore an actigraph (armband and watch communicator) that provided instantaneous PA feedback (steps taken and kilocalories) and sleep parameters (duration and efficiency). EMA compliance rates for the armband and phone calls were 74.7 +/- 0.3% and 64.2 +/- 0.3%, respectively. Data from the armband and phone calls are presented to illustrate the depth of information acquired by utilizing this innovative methodology. PMID- 20019676 TI - Family physicians' barriers to cancer screening in extremely obese patients. AB - Extremely obese women are less likely than nonobese women to receive breast and cervical cancer screening examinations. Reasons for this disparity are unclear and may stem from patient and/or physician barriers. This sequential mixed methods study used individual in-depth interviews of 15 family physicians followed by a mail survey of 255 family physicians (53% response rate) to understand the barriers they faced in performing cancer screening examinations in extremely obese women. Barriers fell into three main areas: (i) difficulty doing pelvic and breast exams; (ii) inadequate equipment; and (iii) challenges overcoming patient barriers and refusal. This led some physicians to avoid performing breast and pelvic examinations on extremely obese women. Having more knowledge about specific examination techniques was associated with less difficulty in palpating lumps on breast and pelvic examinations (P < 0.005). Physicians perceived that embarrassment, aversion to undressing, and avoidance of discussions related to their weight were the most frequent barriers extremely obese women had with getting physical examinations. Educating and/or motivating patients and addressing fears were strategies used most frequently when patients refused mammograms or Pap smears. Interventions focusing on physician barriers, such as educating them on specific examination techniques, obtaining adequate equipment and supplies, and providing resources to assist physicians in dealing with patient barriers and refusal, may be fruitful in increasing cancer screening rates in extremely obese patients. Future research studies testing the effectiveness of these strategies are needed to improve cancer outcomes in this high-risk population. PMID- 20019677 TI - Effects of a low-intensity intervention that prescribed a low-carbohydrate vs. a low-fat diet in obese, diabetic participants. AB - Low-carbohydrate diets have been associated with significant reductions in weight and HbA(1c) in obese, diabetic participants who received high-intensity lifestyle modification for 6 or 12 months. This investigation sought to determine whether comparable results to those of short-term, intensive interventions could be achieved over a 24-month study period using a low-intensity intervention that approximates what is feasible in outpatient practice. A total of 144 obese, diabetic participants were randomly assigned to a low-carbohydrate diet (<30 g/day) or to a low fat diet (or=3.0 (in mg/dl) is a marker of insulin resistance in overweight persons. We aimed at assessing cardiovascular risk profile in 301 overweight elderly Neapolitan outpatients, according to TG/HDL ratio and metabolic syndrome (MS), diagnosed by National Cholesterol Education Program (NCEP) and International Diabetes Federation (IDF) criteria. TG/HDL ratio was >or=3.0 in 97 patients (group A) and <3.0 in 204 (group B). Overall, 93-97% of group A patients and 38-51% of group B patients had MS, depending on the diagnostic criterion. Group A patients with MS had significantly higher waist-to-hip ratio, total and non-HDL cholesterol than group B patients with MS. In group B, MS and non-MS patients had similar waist-to-hip ratio, blood pressure, total and non-HDL cholesterol. Ten year coronary risk, calculated by the Framingham equations (n = 243), was 10.3 +/- 5% in group B, non MS patients; 13.1 +/- 6% in group B, MS patients; 19.9 +/- 8% in group A (F = 32.8; P < 0.001). At the multiple regression analysis, TG/HDL ratio was associated with coronary risk (r(2) = 0.227) more closely than gender, blood pressure, waist-to-hip ratio, non HDL cholesterol, and MS considered as a whole. A separate regression analysis showed that the logarithmically transformed TG/HDL ratio, an index of the HDL cholesterol esterification rate, is also associated with coronary risk (r(2) = 0.252). Thus, TG/HDL ratio could help to characterize high-risk overweight patients deserving a special therapeutic effort. Cardiovascular risk profile of insulin-sensitive patients, identified by lower values of this parameter, is only moderately affected by MS. PMID- 20019685 TI - Interleukin-15 contributes to the regulation of murine adipose tissue and human adipocytes. AB - An alarming global rise in the prevalence of obesity and its contribution to the development of chronic diseases is a serious health concern. Recently, obesity has been described as a chronic low-grade inflammatory condition, influenced by both adipose tissue and immune cells suggesting proinflammatory cytokines may play a role in its etiology. Here we examined the effects of interleukin-15 (IL 15) on adipose tissue and its association with obesity. Over expression of IL-15 (IL-15tg) was associated with lean body condition whereas lack of IL-15 (IL-15(-/ )) results in significant increase in weight gain without altering appetite. Interestingly, there were no differences in proinflammatory cytokines such as IL 6 and tumor necrosis factor-alpha (TNF-alpha) in serum between the three strains of mice. In addition, there were significant numbers of natural killer (NK) cells in fat tissues from IL-15tg and B6 compared to IL-15(-/-) mice. IL-15 treatment results in significant weight loss in IL-15(-/-) knockout and diet-induced obese mice independent of food intake. Fat pad cross-sections show decreased pad size with over expression of IL-15 is due to adipocyte shrinkage. IL-15 induces weight loss without altering food consumption by affecting lipid deposition in adipocytes. Treatment of differentiated human adipocytes with recombinant human IL-15 protein resulted in decreased lipid deposition. In addition, obese patients had significantly lower serum IL-15 levels when compared to normal weight individuals. These results clearly suggest that IL-15 may be involved in adipose tissue regulation and linked to obesity. PMID- 20019686 TI - Sex differences in molecular neuroscience: from fruit flies to humans. AB - A plethora of discoveries relating to sex influences on brain function is rapidly moving this field into the spotlight for most areas of neuroscience. The domain of molecular or genetic neuroscience is no exception. The goal of this article is to highlight key developments concerning sex-based dimorphisms in molecular neuroscience, describe control mechanisms regulating these differences, address the implications of these dimorphisms for normal and abnormal brain function and discuss what these advances mean for future work in the field. The overriding conclusion is that, as for neuroscience in general, molecular neuroscience has to take into account potential sex influences that might modify signalling pathways. PMID- 20019687 TI - Kinase mutations in human disease: interpreting genotype-phenotype relationships. AB - Protein kinases are one of the largest families of evolutionarily related proteins and comprise one of the most abundant gene families in humans. Here we survey kinase gene mutations from the perspective of human disease phenotypes and further analyse the structural features of mutant kinases, including mutational hotspots. Our evaluation of the genotype-phenotype relationship across 915 human kinase mutations - that underlie 67 single-gene diseases, mainly inherited developmental and metabolic disorders and also certain cancers - enhances our understanding of the role of kinases in development, kinase dysfunction in pathogenesis and kinases as potential targets for therapy. PMID- 20019689 TI - Close encounters: reflections on the successes and near misses of exposure science. PMID- 20019691 TI - Drug transporters: recent advances and therapeutic applications. PMID- 20019688 TI - RNA processing and its regulation: global insights into biological networks. AB - In recent years views of eukaryotic gene expression have been transformed by the finding that enormous diversity can be generated at the RNA level. Advances in technologies for characterizing RNA populations are revealing increasingly complete descriptions of RNA regulation and complexity; for example, through alternative splicing, alternative polyadenylation and RNA editing. New biochemical strategies to map protein-RNA interactions in vivo are yielding transcriptome-wide insights into mechanisms of RNA processing. These advances, combined with bioinformatics and genetic validation, are leading to the generation of functional RNA maps that reveal the rules underlying RNA regulation and networks of biologically coherent transcripts. Together these are providing new insights into molecular cell biology and disease. PMID- 20019694 TI - Transporters in drug-refractory epilepsy: clinical significance. AB - Drug resistance remains an unmet challenge in a variety of neurological disorders, but epilepsy is probably the refractory disease that has received most experimental, preclinical, and therapeutic attention. Although resective surgery continues to improve our ability to provide seizure relief, new discoveries have potential as alternative therapeutic approaches to multiple drug resistance. As discussed here, the field is replete with controversies and false starts, in particular as it concerns the existence of genetic predisposition to inadequate pharmacological seizure control. PMID- 20019695 TI - ABC transporters in drug-refractory epilepsy: limited clinical significance of pharmacogenetics? PMID- 20019696 TI - A piece in the puzzle of personalized medicine. AB - Predicting individual drug response based on inherited factors remains a major challenge. Some examples of significant contributions of single genetic variants to overall therapeutic success of drugs in complex diseases are currently emerging. For most compounds, however, multiple genetic and nongenetic factors will modify drug action. Comprehensive integration of these factors will determine the role of pharmacogenomics for personalized medicine. PMID- 20019697 TI - Biomarker qualification via public-private partnerships. AB - Biomarkers linked to patient outcomes (safety and efficacy) have an increasingly important role in drug development. Consequently, validation and qualification of such biomarkers are essential, often requiring large data sets from well controlled randomized clinical trials. In the December 2009 issue of Clinical Pharmacology & Therapeutics, investigators utilizing data from four pharmaceutical companies and working under the auspices of the Biomarkers Consortium described the utility of adiponectin as an early predictor of glycemic control in diabetic patients taking peroxisome proliferator-activated receptor (PPAR) agonists. This work illustrates the advantages of large public-private partnerships for biomarker qualification. PMID- 20019698 TI - Quantification of T-cell proliferation for individualizing immunosuppressive therapy for transplantation patients. AB - The serious side effects and complications related to the lifelong use of immunosuppressors in organ transplantation have fueled research into their possible minimization. Immunosuppressive therapy in organ transplantation is therefore tentatively moving from a phase of empirical administration toward individualized therapy. This process is highly dependent on the development of monitoring methods to detect individual immune states. The results of the studies by Kurata et al. support the usefulness of pharmacodynamic assays of lymphocyte function for the predictive monitoring of the effects of immunosuppressive drug therapy. PMID- 20019699 TI - Expediting patients' access to medicines by improving the predictability of drug development and the regulatory approval process. AB - Ideally, well-designed global pharmaceutical development programs that include simultaneous submissions to multiple regulatory agencies can result in predictable regulatory approvals and expedited access to medicines for all patients. The workshops described herein investigated current trends in development and submission strategies along with regulatory review performance data in order to consider whether barriers to predictable expedited approval outcomes can be overcome through innovative clinical development approaches and a better understanding of review processes and procedures, particularly as these relate to the assessment of a product's risk-benefit profile. PMID- 20019700 TI - The International Transporter Consortium: a collaborative group of scientists from academia, industry, and the FDA. AB - The US Food and Drug Administration-led Critical Path Initiative, launched in 2004, has resulted in an array of activities focused on the sciences that support the development of human medical products.(1) These activities include the development of new scientific tools, such as in vitro testing, qualified biomarkers of drug safety, and innovative new methods in study design and data analysis.(2) As a result of the Critical Path Initiative and enormous advances in the field of membrane transporters, the International Transporter Consortium was formed. PMID- 20019701 TI - Vasculoprotective effect of cilostazol in aldosterone-induced hypertensive rats. AB - Cilostazol (CILO), a selective inhibitor of phosphodiesterase 3 with potent antithrombotic property, has been shown to have a vasculoprotective effect in atherosclerosis animal models due to its potential anti-inflammatory and antioxidant actions. This study was undertaken to investigate whether CILO has in fact any vasculoprotective effects in aldosterone-induced hypertensive rats (Aldo rats), and whether CILO affects Aldo-induced oxidative stress, nitric oxide (NO) production and pro-inflammatory gene expression. Treatment with CILO markedly ameliorated perivascular inflammatory changes in the coronary arterioles of Aldo rats without affecting the systolic blood pressure and left ventricular weight. Treatment with CILO also prevented the increase in plasma levels of thiobarbituric acid-reactive substances, an oxidative stress marker, as well as decreased urinary NOx excretion in Aldo-rats. Furthermore, CILO almost completely inhibited a set of upregulated proinflammatory genes (ICAM-1, MCP-1, PDGF-A, osteopontin, MMP-2 and ACE), as well as NAD(P)H oxidase components (p22phox, gp91phox, p47phox) and Aldo-inducible genes (SGK-1 and NHE-1) in the aortic tissues from Aldo-rats. Taken together, this study showed for the first time that CILO prevented Aldo-induced vascular inflammation and injury without affecting the blood pressure, suggesting its vasculoprotective effect on Aldo-induced vascular injury independent of blood pressure. PMID- 20019702 TI - Association of eNOS gene polymorphism (rs3918166) with blood pressure in adult Japanese. PMID- 20019703 TI - Possible roles of human (pro)renin receptor suggested by recent clinical and experimental findings. AB - Numerous in vitro and in vivo animal studies using the (pro)renin receptor (P)RR blocker handle region peptide have suggested an important role of (P)RR in the pathogenesis of end-stage organ damage in patients with diabetes and hypertension. In addition, a limited number of clinical studies have suggested an association between (P)RR gene polymorphisms and blood pressure levels and between (P)RR mRNA levels and angiotensin-converting enzyme mRNA levels in human arteries. However, recent studies have shown that the (P)RR is divided into its soluble form and a residual hydrophobic part, which includes ATPase 6 associated protein 2, within cells. Therefore, the (P)RR may have a more complex function than previously thought. In addition, the physiological roles of the (P)RR remain undetermined, because the construction of (P)RR null mice has not been successful. As a next step for research in this area, a method for determining the soluble (P)RR levels in plasma and urine and the construction of tissue specific (P)RR-knockout mice are needed to elucidate the roles of the (P)RR in physiology and pathophysiology. PMID- 20019704 TI - Does ACE2 contribute to the development of hypertension? PMID- 20019705 TI - Dental or dentist? PMID- 20019707 TI - Clarifying points. PMID- 20019708 TI - Perspective on dentistry. PMID- 20019709 TI - Back door dentistry. PMID- 20019710 TI - Averting damage. PMID- 20019711 TI - Folk remedy. PMID- 20019712 TI - Disposed to adipose. PMID- 20019713 TI - Government barring scheme list will affect dentists. PMID- 20019714 TI - Salford Faculty Dean visits Hong Kong health unit. PMID- 20019716 TI - Prizes in risk management competition. PMID- 20019718 TI - Call for dentists to ensure they are using the safest products. PMID- 20019719 TI - Royal recognition for Cardiff. PMID- 20019726 TI - Evaluation of an NHS practice-based minor oral surgery service. AB - OBJECTIVES: To evaluate a pilot specialist practice-based minor oral surgery service. METHODS: Service monitoring data were analysed to evaluate activity, waiting times and outcomes. Patient satisfaction was assessed by a modified version of the Dental Visit Satisfaction Scale. RESULTS: Of 705 treatment appointments, 12.7% were not attended and treatment was not provided in another 11.6%. Mean waiting time from referral receipt to treatment was 6.8 weeks. Treatment provided included surgical removal of third molars, surgical removal of non-third molars and surgical endodontics (26.7%, 60.4% and 4.9% of cases, respectively). Antibiotics were prescribed in 15.0% of treatment cases and 2.3% of treatment cases required appointments for postoperative complications. The response rate for the satisfaction survey was 81%. All participants reported overall satisfaction and strongly agreed/agreed with positive attitudinal statements about the oral surgeon's communication/information giving, technical competence and understanding/acceptance. 77.8% were seen on time and 22.2% within 15 minutes of their appointment. Overall 74.1% felt the standard of service was better than they would expect from a hospital and none felt it was worse. CONCLUSION: The findings of the evaluation suggest that specialist minor oral surgery can be successfully provided in dental practice and is acceptable to patients. PMID- 20019727 TI - Dental patients' use of the Internet. AB - AIMS AND OBJECTIVES: To determine the use of the Internet by patients attending a range of dental clinics to search for information regarding dental procedures, and also to investigate their interest in online dental consultations and 'dental tourism'. METHODS: A questionnaire was designed and randomly distributed to 520 patients attending the restorative dentistry, dental surgery and oral medicine clinics of Cork University Dental School and Hospital. RESULTS: Of the 520 questionnaires distributed, 500 were completed leading to a response rate of 96.2%. The majority of patients were familiar with using the Internet on a daily basis, with only 163 (32.6%) patients not using the Internet in their everyday lives. One hundred and seventy-seven (34.5%) patients either researched their presenting dental/oral condition or had a family or friend research their condition on their behalf. One hundred and eighty-five (37%) patients would consult with a dental practitioner online regarding an oral problem and a similar number (n=186) of patients surveyed would consider using the Internet to plan trips abroad for dental treatment. CONCLUSION: Practitioner-led direction for patients regarding quality information sources online is important. With the increased interest in travelling abroad for dental treatment, guidance for patients and practitioners regarding the legal and ethical issues pertaining to dental tourism is critical. PMID- 20019744 TI - Molecular mechanisms of late apoptotic/necrotic cell clearance. AB - Phagocytosis serves as one of the key processes involved in development, maintenance of tissue homeostasis, as well as in eliminating pathogens from an organism. Under normal physiological conditions, dying cells (e.g., apoptotic and necrotic cells) and pathogens (e.g., bacteria and fungi) are rapidly detected and removed by professional phagocytes such as macrophages and dendritic cells (DCs). In most cases, specific receptors and opsonins are used by phagocytes to recognize and bind their target cells, which can trigger the intracellular signalling events required for phagocytosis. Depending on the type of target cell, phagocytes may also release both immunomodulatory molecules and growth factors to orchestrate a subsequent immune response and wound healing process. In recent years, evidence is growing that opsonins and receptors involved in the removal of pathogens can also aid the disposal of dying cells at all stages of cell death, in particular plasma membrane-damaged cells such as late apoptotic and necrotic cells. This review provides an overview of the molecular mechanisms and the immunological outcomes of late apoptotic/necrotic cell removal and highlights the striking similarities between late apoptotic/necrotic cell and pathogen clearance. PMID- 20019745 TI - Nurturing brain plasticity: impact of environmental enrichment. AB - Environmental enrichment (EE) is known to profoundly affect the central nervous system (CNS) at the functional, anatomical and molecular level, both during the critical period and during adulthood. Recent studies focusing on the visual system have shown that these effects are associated with the recruitment of previously unsuspected neural plasticity processes. At early stages of brain development, EE triggers a marked acceleration in the maturation of the visual system, with maternal behaviour acting as a fundamental mediator of the enriched experience in both the foetus and the newborn. In adult brain, EE enhances plasticity in the cerebral cortex, allowing the recovery of visual functions in amblyopic animals. The molecular substrate of the effects of EE on brain plasticity is multi-factorial, with reduced intracerebral inhibition, enhanced neurotrophin expression and epigenetic changes at the level of chromatin structure. These findings shed new light on the potential of EE as a non-invasive strategy to ameliorate deficits in the development of the CNS and to treat neurological disorders. PMID- 20019746 TI - Akt2-mediated phosphorylation of Pitx2 controls Ccnd1 mRNA decay during muscle cell differentiation. AB - Paired-like homeodomain 2 (Pitx2), first identified as the gene responsible for the Axenfeld-Rieger syndrome, encodes a protein factor that, controlling cell proliferation in a tissue-specific manner, has a crucial role in morphogenesis. During embryonic development, Pitx2 exerts a role in the expansion of muscle progenitors and is expressed at all stages of myogenic progression. In this study, we show that Pitx2 is phosphorylated by the protein kinase Akt2 and is necessary to ensure proper C2C12 myoblast proliferation and differentiation. Pitx2 associates with a ribonucleoprotein complex that includes the mRNA stabilizing factor HuR and sustains Ccnd1 (also known as Cyclin D1) expression, thereby prolonging its mRNA half-life. When the differentiation program is initiated, phosphorylation by Akt2 impairs the ability of Pitx2 to associate with the Ccnd1 mRNA-stabilizing complex that includes HuR and, as a consequence, Ccnd1 mRNA half-life is shortened. We propose that unphosphorylated Pitx2 is required to favor HuR-mediated Ccnd1 mRNA stabilization, thus sustaining myoblast proliferation. Upon Akt2-phosphorylation, the complex Pitx2/HuR/Ccnd1 mRNA dissociates and Ccnd1 mRNA is destabilized. These events contribute to the switch of C2C12 cells from a proliferating to a differentiating phenotype. PMID- 20019747 TI - Apoptosis and the yeast actin cytoskeleton. AB - Actin represents one of the most abundant and extensively studied proteins found in eukaryotic cells. It has been identified as a major target for destruction during the process of apoptosis. Recent research has also highlighted a role for cytoskeletal components in the initiation and inhibition of apoptotic processes. The high degree of conservation that exists between actins from divergent eukaryotes, particularly with respect to those that contribute to the cytoskeleton, has meant that functional studies from the model yeast Saccharomyces cerevisiae have proven useful in elucidating its cellular roles. Within the context of apoptosis in yeasts, actin seems to function as part of the signalling mechanisms that link nutritional sensing to a mitochondrial-dependent commitment to cell death. Studies in yeasts have also shown that oxidative damage accrued by the actin cytoskeleton is closely monitored and is tethered to an apoptotic response. Strong, but as yet, undefined links between the actin cytoskeleton and apoptosis have also been described in studies from plant and animal systems. The widespread involvement of actin in apoptotic mechanisms from diverse eukaryotic organisms raises the possibility of conserved regulatory pathways, further strengthening the relevance of yeast research in this area. PMID- 20019748 TI - Proapoptotic signalling through Toll-like receptor-3 involves TRIF-dependent activation of caspase-8 and is under the control of inhibitor of apoptosis proteins in melanoma cells. AB - Toll-like receptor-3 (TLR3), a member of an immune recognition receptor family, is widely expressed in tumour cells and has been shown previously to have the capacity to not only activate immune signalling pathways, but also to exert proapoptotic activity in some cells. We show here that HaCaT human keratinocytes are susceptible to apoptosis induction by the TLR3 ligand poly I:C, and use these cells as a model to analyse the apoptotic signalling pathway. Although the BH3 only protein Noxa was transcriptionally induced by poly I:C and translocated to mitochondria, RNAi experiments showed that the BH3-only proteins Noxa, Bim and Puma were individually dispensable for poly I:C-induced apoptosis. Instead, poly I:C-induced activation of caspase-8 via TLR3 and its adapter TRIF was required for apoptosis. In human melanoma cell lines poly I:C failed to induce apoptosis unless protein synthesis was blocked. Significantly, sensitisation towards poly I:C-dependent caspase-8 activation and apoptosis in melanoma cells was also achieved by the synthetic Smac mimetic/inhibitor of apoptosis protein (IAP) antagonist, LBW242, or by specific downregulation of cIAP1 by siRNA. Inactivation of caspase-8 by CrmA overexpression reduced poly I:C/LBW242-induced apoptosis. These results indicate that the proapoptotic activity of TLR3/TRIF/caspase-8 in melanoma cells is under the control of IAPs, and the use of novel Smac mimetics might be a feasible approach to target melanoma. PMID- 20019749 TI - Mesenchymal stem cells improve small intestinal integrity through regulation of endogenous epithelial cell homeostasis. AB - Patients who undergo pelvic or abdominal radiotherapy may develop acute and/or chronic side effects resulting from gastrointestinal tract (GIT) alterations. In this study, we address the question of the regenerative capability of mesenchymal stem cells (MSC) after radiation-induced GIT injury. We also propose cellular targets of MSC therapy. We report that the infusion of human bone marrow-derived MSC (hMSC) provides a therapeutic benefit to NOD/SCID mice undergoing radiation induced GIT failure. We observed that hMSC treatment brings about fast recovery of the small intestine (structure and function) in mice with reversible alterations and extends the life of mice with irreversible GIT disorders. The effects of hMSC are a consequence of their ability to improve the renewal capability of small intestinal epithelium. hMSC treatment favors the re establishment of cellular homeostasis by both increasing endogenous proliferation processes (Ki67 immunostaining) and inhibiting apoptosis (TUNEL staining) of radiation-induced small intestinal epithelial cells. Our results suggest that MSC infusion may be used as a therapeutic treatment to limit radiation-induced GIT damage. PMID- 20019750 TI - Monoallelic but not biallelic loss of Dicer1 promotes tumorigenesis in vivo. AB - Human tumors are characterized by widespread reduction in microRNA (miRNA) expression, although it is unclear how such changes come about and whether they have an etiological role in the disease. Importantly, miRNA knockdown has been shown to enhance the tumorigenic potential of human lung adenocarcinoma cells. A defect in miRNA processing is one possible mechanism for global downregulation. To explore this possibility in more detail in vivo, we have manipulated Dicer1 gene dosage in a mouse model of retinoblastoma. We show that although monoallelic loss of Dicer1 does not affect normal retinal development, it dramatically accelerates tumor formation on a retinoblastoma-sensitized background. Importantly, these tumors retain one wild-type Dicer1 allele and exhibit only a partial decrease in miRNA processing. Accordingly, in silico analysis of human cancer genome data reveals frequent hemizygous, but not homozygous, deletions of DICER1. Strikingly, complete loss of Dicer1 function in mice did not accelerate retinoblastoma formation. miRNA profiling of these tumors identified members of the let-7 and miR-34 families as candidate tumor suppressors in retinoblastoma. We conclude that Dicer1 functions as a haploinsufficient tumor suppressor. This finding has implications for cancer etiology and cancer therapy. PMID- 20019751 TI - Yeast unfolds the road map toward alpha-synuclein-induced cell death. AB - The budding yeast Saccharomyces cerevisiae has contributed significantly to our current understanding of eukaryotic cell biology. It served as a tool and model for unraveling the molecular basis of a wide variety of cellular phenomena, which seem to be conserved in other organisms. During the last decade, yeast has also extensively been used to study the mechanisms underlying several human diseases, including age-associated neurodegenerative disorders, such as Parkinson's, Huntington's and Alzheimer's disease. In this review, we focus on a yeast model for synucleinopathies and summarize recent studies that not only provided new clues on how the misfolding of alpha-synuclein (alpha-syn) triggers toxicity and eventually cell death, but that also led to the identification of conserved suppressor proteins, which are effective in protecting cells, including neurons, from the alpha-syn-induced cytotoxicity. PMID- 20019752 TI - Inheritance of a novel mutated allele of the OCA2 gene associated with high incidence of oculocutaneous albinism in a Polynesian community. AB - Oculocutaneous albinism type 2 (OCA2) is a human autosomal-recessive hypopigmentation disorder associated with pathological mutations of the OCA2 gene. In this study, we investigated a form of OCA in a Polynesian population with an observed phenotype characterized by fair skin, some brown nevi present in the sun-exposed areas and green or blue eyes. Hair presented with a unique red coloration since birth, with tones ranging across individuals from Yellow-Red to Brown-Red, or Auburn. We genetically screened for mutations in the OCA2 and MC1R genes as their products have previously been shown to be associated with red hair/fair skin and OCA2. The SLC45A2 gene was also screened to identify any possible relation to skin color variation. We have identified a novel missense substitution in the OCA2 gene (Gly775Asp) responsible for OCA2 in individuals of Polynesian heritage from Tuvalu. The estimated incidence of this form of OCA2 in the primary study community is believed to occur at one of the highest recorded rates of albinism at approximately 1 per 669 individuals. In addition, we have analyzed four unrelated individuals with albinism who have Polynesian heritage from three other separate communities and found they carry the same OCA2 mutation. We also analyzed an out-group comprising three unrelated individuals with albinism of Melanesian ancestries from two separate communities, one Australian Aboriginal and three Australian Caucasians, and did not detect this mutation. We hypothesize that this mutation may be Polynesian specific and that it originated from a common founder. PMID- 20019753 TI - Atrial fibrillation: Dronedarone and amiodarone-the safety versus efficacy debate. PMID- 20019754 TI - Device therapy: Indications for ICD-CRT in mildly symptomatic heart failure. PMID- 20019755 TI - Intervention: The AMADEUS study in perspective. PMID- 20019756 TI - Atrial fibrillation: A promising new anticoagulant for stroke prevention. PMID- 20019757 TI - Mitochondrial dynamics regulate the RIG-I-like receptor antiviral pathway. AB - The intracellular retinoic acid-inducible gene I-like receptors (RLRs) sense viral ribonucleic acid and signal through the mitochondrial protein mitochondrial antiviral signalling (MAVS) to trigger the production of type I interferons and proinflammatory cytokines. In this study, we report that RLR activation promotes elongation of the mitochondrial network. Mimicking this elongation enhances signalling downstream from MAVS and favours the binding of MAVS to stimulator of interferon genes, an endoplasmic reticulum (ER) protein involved in the RLR pathway. By contrast, enforced mitochondrial fragmentation dampens signalling and reduces the association between both proteins. Our finding that MAVS is associated with a pool of mitofusin 1, a protein of the mitochondrial fusion machinery, suggests that MAVS is capable of regulating mitochondrial dynamics to facilitate the mitochondria-ER association required for signal transduction. Importantly, we observed that viral mitochondria-localized inhibitor of apoptosis, a cytomegalovirus (CMV) antiapoptotic protein that promotes mitochondrial fragmentation, inhibits signalling downstream from MAVS, suggesting a possible new immune modulation strategy of the CMV. PMID- 20019758 TI - Transfer-messenger RNA controls the translation of cell-cycle and stress proteins in Streptomyces. AB - The transfer-messenger RNA (tmRNA)-mediated trans-translation mechanism is highly conserved in bacteria and functions primarily as a system for the rescue of stalled ribosomes and the removal of aberrantly produced proteins. Here, we show that in the antibiotic-producing soil bacterium Streptomyces coelicolor, trans translation has a specialized role in stress management. Analysis of proteins that were carboxy-terminally His(8)-tagged by a recombinant tmRNA identified only 10 targets, including the stress proteins: DnaK heat-shock protein 70, thiostrepton-induced protein A, universal stress protein A, elongation factor Tu3, and the cell-cycle control proteins DasR, SsgA, SsgF and SsgR. Although tmRNA-tagged proteins are degraded swiftly, the translation of dnaK and dasR messenger RNAs (mRNAs) depends fully on tmRNA, whereas transcription is unaffected. The data unveil a surprisingly dedicated functionality for tmRNA, promoting the translation of the same mRNA it targets, at the expense of sacrificing the first nascent protein. In streptomycetes, tmRNA has evolved into a dedicated task force that ensures the instantaneous response to the exposure to stress. PMID- 20019759 TI - Efficacy and safety of fixed combinations of latanoprost/timolol and dorzolamide/timolol in open-angle glaucoma or ocular hypertension. AB - AIMS: To compare intraocular pressure (IOP) reductions with fixed-combination (FC) latanoprost/timolol once daily in the evening vsFC dorzolamide/timolol twice daily. METHODS: This evaluator-masked, multicentre, controlled clinical trial randomized subjects with primary open-angle glaucoma or ocular hypertension with IOP insufficiently responsive to beta-blocker therapy (screening IOP>21 and <37 mm Hg) to FC latanoprost-timolol (N=135) or FC dorzolamide/timolol (N=135). At screening, baseline, and after 4 and 12 weeks of therapy, IOP was measured three times at 0800, 1200, and 1600 hours. Adverse events were recorded at each visit. The primary efficacy end point was whether either FC could be shown to be inferior to the other with respect to change in mean daytime IOP from baseline to week 12. RESULTS: Mean daytime IOP levels were similar at baseline. Mean reductions in daytime IOP from baseline to week 12 were -9.7 mm Hg for FC latanoprost-timolol and -9.5 mm Hg for FC dorzolamide/timolol. The difference between FC latanoprost/timolol-FC dorzolamide-timolol was -0.2 mm Hg (95% confidence interval (CI), -0.8 to -0.4 mm Hg). The upper bound of the 95% CI was <1.5 mm Hg, indicating that neither FC is inferior to the other. However, a significantly greater percentage of subjects treated with FC latanoprost/timolol achieved IOP levels or =75%, 4. RESULTS: There were a variety of degrees of thickening or splitting GBM in patients with Alport's syndrome. Laminin alpha1 was positive in glomerular mesangial area and absolutely negative in GBM and laminin alpha5 was evenly positive in GBM in normal tissue. In Alport's syndrome, laminin alpha1 was much weaker in glomerular mesangial area, but strongly positive in GBM; laminin alpha5 in GBM was prominently reduced. There was a high negative correlation of semiquantitative scores between laminin alpha1 and laminin alpha5 (r=-0.83, P<0.001). The extension of thickening or splitting GBM was positively correlated with scores of laminin alpha1 in GBM (r=0.76, P<0.001; r=0.56, P=0.015), and was negatively correlated with scores of laminin alpha5 in GBM (r=-0.59, P=0.010; r=-0.53, P=0.025). CONCLUSION: Abnormal distribution of laminin alpha1 and laminin alpha5 in GBM is correlated with GBM thickening and splitting in human Alport's syndrome. PMID- 20019772 TI - [Role of transforming growth factor beta1 in the development of atrophic gastritis]. AB - OBJECTIVE: To investigate the role of transforming growth factor beta1 (TGFbeta1) in the development of Helicobacter pylori (H.pylori)-associated non-metaplastic atrophic gastritis. METHODS: The expressions of TGFbeta1, CD68 and smooth muscle actin(SMA) were detected immunohistochemically in 10 patients with mild non atrophic gastritis, 30 patients with mild non-metaplastic atrophic gastritis, and 32 patients with severe non-metaplastic atrophic gastritis having H.pylori infecion. Meanwhile, three cases of mild non-atrophic gastritis and 4 cases of severe non-metaplastic atrophic gastritis were observed with electron microscope. RESULTS: The count of TGFbeta1 positive cells per high-power field (HPF) in severe non-metaplastic atrophic gastritis group (53 +/- 22) was significantly higher than that in mild non-atrophic gastritis group(22+/-9/HPF) and mild non atrophic gastritis group(0-3/HPF, P<0.01). The count of CD68 positive cells in severe non-metaplastic atrophic gastritis group (23+/-7/HPF) was significantly higher than that in mild non-atrophic gastritis group (13+/-6/HPF) and mild non atrophic gastritis group(0-3/HPF, P<0.01). Correlation analysis showed that the expressions of TGFbeta1 and CD68 had a moderate correlation in each group (r=0.634, P<0.01; r=0.699, P<0.01). Compared with mild non-atrophic gastritis, SMA-positive myofibroblasts and smooth muscle cells in the lamina propria increased in mild and severe non-metaplastic atrophic gastritis. Ultrastructurally, the proliferation of fibroblasts in gastric lamina propria was observed in mild non-atrophic gastritis, while the proliferation of fibroblasts and presence of myofibroblasts could be observed in mild non-metaplastic atrophic gastritis, and there was a parallel phenomenon between myofibroblasts and fibroblasts, as well as smooth muscle cells. CONCLUSION: Our findings indicate that TGFbeta1 expression increases with severity of H.pylori- associated non metaplastic atrophic gastritis, suggesting that TGFbeta1 might play an important role in the development of non-metaplastic atrophic gastritis. PMID- 20019773 TI - [Anticancer effects of a novel indolinone compound in nasopharyngeal carcinoma cells]. AB - OBJECTIVE: To develop novel targeted anticancer medicines for effective treatment of nasopharyngeal carcinoma (NPC), a prevalent malignant disease in southern China and southeast Asia. METHODS: CNE cells were treated with a novel indolinone IF239 synthesized by our research group. Cell viability was determined by the acid phosphatase assay (APA). Morphologic changes and adhesion status of CNE cells treated with IF239 were observed under a light microscope. Flow cytometry was used to analyze the cell cycle phases . Key regulating molecules in the cell cycle progression were detected by Western blotting. RESULTS: IF239 had potent cytotoxic effect on CNE cells. The possible antitumor mechanisms of IF239 involved inhibition of cell adhesion and cell cycle arrest in the G2/M phase. Moreover, G2/M arrest caused by IF239 was related to up-regulation of both cyclin B1 and the phosphorylation level of CDK1. CONCLUSION: IF239 has high anticancer activity over CNE cells, and has unique anticancer mechanisms, suggesting that IF239 has promising application potentials. PMID- 20019774 TI - [High-risk factors and clinical pathological analysis in 21 cases of fatal pulmonary embolism]. AB - OBJECTIVE: To explore the clinical and pathological characteristics of fatal pulmonary thromboembolism, especially the contribution of acquired and genetic risk factors of fatal pulmonary thromboembolism, so as to provide some useful information for its clinical prevention and treatment. METHODS: The medical dispute autopsy cases performed at the Department of Pathology, School of Basic Medical Sciences, Peking University from January 1, 2002 to September 30, 2008 were retrospectively reviewed. FV ( factor V ) Leiden mutation and FII ( factor II) G20210A mutation were analyzed by using methods of PCR and restriction fragment length polymorphism (RFLP) in fatal pulmonary embolism cases and random selected control cases of adult autopsy. RESULTS: There were 21 patients who died from fatal pulmonary thromboembolism, accounting for 12.96% (21/162) of adult autopsy cases. In these cases, there was at least one of the known risk factors in each case and more than two risk factors could be found in most cases. The acquired risk factors, such as surgery, trauma and fractures, cancer, pregnancy and puerperium, the elderly, hypertension, diabetes, infections, could be found in these patients. FV Leiden mutation and FII G20210A mutation were not detected in any of the 21 cases of fatal pulmonary thromboembolism cases and six control cases. Of all the 21 cases, quantitative D-dimer measurement was performed only in six patients and D-dime level in these six samples was all significantly increased. According to the records, there was only 1 patient treated by preventive anticoagulants in the 21 patients. CONCLUSION: Fatal pulmonary thromboembolism is an important reason for medical dispute. Fractures and trauma, surgery and other acquired factors are the main reasons for fatal pulmonary embolism in Chinese. Genetic factors (FV Leiden and FII G20210A mutations) are not the reason for Chinese to suffer from fatal pulmonary thromboembolism. Assessment of risk factors (especially traumatic fractures and surgical intervention) of pulmonary thromboembolism may play an important role in taking appropriate preventive procedures, early diagnosis and reasonable treatment to reduce its mortality. PMID- 20019775 TI - [Clinical pathological analysis in 14 cases of pancreatic solid-pseudopapillary tumors]. AB - OBJECTIVE: To observe clinical and pathological features of pancreatic solid pseudopapillary tumor (SPPT), and to find some useful immunohistochemical methods for its differential diagnosis. METHODS: The clinical features of 14 SPPT patients were obtained. Each case underwent microscopic observation and immunohistochemical staining. The primary antibodies were CgA, Syn, E-cadherin, beta-catenin and Cyclin D1. These results were compared with 5 pancreatic well differentiated tumors and well-differentiated carcinomas (WET/WEC). RESULTS: SPPT mainly involved young women, and the head of pancreas was the commonest location. Tumors were always in solid and cystic gross appearance. Although the tumor's borderlines seemed clear, focal infiltrations could often be identified. The histological features of SPPT were similar in some aspects to those of WET/WEC, especially the solid pattern of WET/WEC. Both of them could express CgA and Syn. But all SPPTs lost E-cadherin membranous signals, and even had some nuclear signals(5/14), while all WET/WECs remained the same staining pattern with normal pancreas cells. beta-catenin positive signals in SPPTs were located both in nuclei and plasmas. WET/WECs' positive signals were all in membranes and plasmas, but negative ones in nuclei. Perinuclear dot-like signals could also be seen in the majority cells, which were similar to normal islet cells' staining pattern. SPPTs' nuclear positive rates of Cyclin D1 were usually more than 70% (12/14). WET/WECs' rates were all lower than 30%. CONCLUSION: Comprehensive analysis of patients' clinical, pathological features and immunohistochemistry results, including E-cadherin, beta-catenin and Cyclin D1, was helpful to the diagnosis of SPPT and its differential diagnosis of WET/WEC. PMID- 20019776 TI - [Expression and function of cyclooxygenase-2 in endometrial carcinoma]. AB - OBJECTIVE: To evaluate the cyclooxygenase-2(COX-2) expression at mRNA and protein levels, as well as its clinical significance, and to study the correlation of COX 2 with angiogenesis, apoptosis and estrogen or progestron receptors. METHODS: Forty-one samples of normal endometrium tissues and 52 ones of endometrial carcinomas(EC) were collected, together with their corresponding clinical information. RT-PCR was adopted to determine the expression of COX-2 mRNA. Immunohistochemical staining was engaged to evaluate the protein expression of COX-2, VEGF, Bcl-2 and the MVD marked by CD105. And TUNEL was used to count the apoptosis cells. RESULTS: (1)Significantly higher mRNA and protein levels of COX 2 were found in endometrial carcinomas than in normal endometrial tissues (P<0.001). (2)At protein level, COX-2 expression was related to the grade (P=0.020), but had nothing to do with the stage, pathological type, depth of myometrium invasion and lymph node metastasis.Neither did the level of COX-2 mRNA with all the factors above. (3)The level of COX-2 mRAN had nothing to do with that of protein(P=0.125, r=0.222). (4)The higher the expression of COX -2 protein was, the higher that of VEG, MVD or Bcl-2 was, and the lower the count of apoptosis cells was P<0.05. (5)The upregulation of COX-2 protein had no correlation with ER-alpha and ER-beta. But it had positive correlation with PRA and PRB, P was 0.031 and 0.007, respectively. CONCLUSION: (1)The expression of COX-2 is significantly higher in EC than in normal endometrium tissue, not only at the mRNA level, but also at protein level. (2)At protein level, COX-2 expression is related to the grade, but has nothing to do with the stage, pathological type, depth of myometrium invasion and lymph node metastasis. Neither does the level of COX-2 mRNA with all the factors above. (3)The upregulation of COX-2 protein in EC correlates with the increase of VEGF protein and MVD. (4)The upregulation of COX-2 protein in EC correlates with the increase of Bcl-2 protein and the decrease of apoptosis cells. (5)The upregulation of COX 2 protein in EC has no significant correlation with ER expression, but has something to do with the increase of PRA and PRB. PMID- 20019777 TI - [Association of SNPs in N-formylpeptide receptor gene with susceptibility of aggressive periodontitis]. AB - OBJECTIVE: To explore the association of the single nucleotide polymorphisms (SNPs) in N-formylpeptide receptor (FPR) gene with the susceptibility of aggressive periodontitis (AgP). METHODS: A total of 94 AgP patients and 73 healthy controls were entered into the study. Peripheral blood sample was obtained from each subject by venepuncture. Genomic DNA was isolated from each sample. The target fragment of FPR gene was amplified by PCR. The SNPs in FPR gene were detected by denature high performance liquid chromatography (DHPLC) combined with DNA sequencing. RESULTS: There were two non-synonymous SNPs in the 370 bp FPR gene fragment: 289C/A and 301G/C. The 289C/A was a novel SNP. No variation in nucleotides 329 and 378 was detected. There were no statistically significant differences in distributions of the genotypes and alleles for FPR289 and FPR301 between AgP patients and healthy controls. Using multivariate logistic regression (adjusted for age and gender), it was showed that the adjusted ORs of AgP for the C(+) genotype and allele C of FPR301 combined with smoking were 5.74 and 5.20 respectively. CONCLUSION: The presence of the C(+) genotype/allele C of FPR301 together with smoking conferred a higher risk for AgP. The result suggests that the SNPs in FPR gene may not be associated with the susceptibility of AgP in Chinese. PMID- 20019778 TI - [Effect of icariin on the mRNA expressions of Cbfalpha1, BMP2, BMP4 in rat osteoblasts]. AB - OBJECTIVE: To investigate the effect of icariin on the proliferation, differentiation, and the mRNA expressions of Cbfalpha1, BMP2, BMP4 of rat osteoblasts. METHODS: Primary rat osteoblastic cells were obtained by sequentia collagenase/trypsin enzyme digestion from calvarial bones of new born (within 24 h) SD rats and were identified by Alkaline phosphatase and alizarin red staining. The passage 3-5 cells were treated with icariin at the concentration of 0 mol/L, 10(-8) mol/L, 10(-7) mol/L, 10(-6) mol/L, 10(-5) mol/L, 10(-4) mol/L for 24 h, 48 h, 72 h, and the proliferation of the cells was measured by CCK-8 assay. The proliferation index was detected by Flow Cytometry and the activity of alkaline phosphatase was determined by p-Nitrophenyl phosphate (pNPP) method after being treated with icariin at the concentration mentioned above for 48 h. The total cellular RNA was extracted 48 h after being treated with icariin at the concentration of 10(-6) mol/L, and the expressions of Cbfalpha1, BMP2, BMP4 mRNA were examined by real-time PCR. RESULTS: Icariin showed no effect on the proliferation of osteoblasts, but improved ALP activity. The Cbfalpha1, BMP2, BMP4 mRNA were significantly upregulated after icariin treatment. CONCLUSION: Icariin could promote the differentiation ability of rat osteoblasts through upregulating the Cbfalpha1, BMP2, BMP4 mRNA expressions. PMID- 20019779 TI - [Clinical characteristics of rheumatoid arthritis with interstitial pulmonary fibrosis]. AB - OBJECTIVE: To analyze the clinical characteristics of interstitial pulmonary fibrosis (IPF) in patients with Rheumatoid arthritis (RA). METHODS: We retrospectively analyzed 198 RA patients with or without IPF. Characteristics of RA-IPF in clinical and lab data were analyzed. Age, duration of disease, clinical and laboratory parameters, history of smoking and medicine were compared between the patients with and without IPF. RESULTS: (1) Among the 198 RA patients, 15.2% (30/198) were found with IPF. 100% RA-IPF patients had HRCT findings. However, 63.3% (19/30) had positive findings in chest X-ray, and only 46.7% (14/30) had the complaints of cough and short breath. Velcro rales were found in 50.0% (15/30) patients with IPF and no acropachy occurred. Only one patient suffered from hypoxemia. IPF presented after the joint symptoms in most patients. (2) RA IPF patients were older than those without IPF [(65.50 +/- 9.71) vs (55.22 +/- 12.98) years, P<0.01]; Higher positivity of anti-keratin antibodies (AKA) were found in RA-IPF compared to patients without IPF (61.5% vs 35.9%, P=0.014). Furthermore, the levels of anti-cyclic citrullinated peptide (CCP) antibody were significantly higher in RA-IPF [(4.38 +/- 2.08) vs (3.20 +/- 2.12), P=0.01]. No differentiation of duration of disease, history of smoking and medicine, IgM rheumatoid factor, IgG rheumatoid factor, anti-nuclear antibody, anti-SSA antibody and levels of immunoglobins and complements were found between the two groups of RA patients with and without IPF. CONCLUSION: The clinical symptoms of IPF in RA patients are mild and more common in older patients. AKA and anti-CCP antibody might be important antibodies associated with RA-IPF. PMID- 20019780 TI - [Clinical features of hypokalemic periodic paralysis]. AB - OBJECTIVE: To explore the clinical features of hypokalemic periodic paralysis, and compare clinical features of primary group with those of thyrotoxicosis secondary group. METHODS: Clinical data of 44 patients with hypokalemic periodic paralysis in Peking University First Hospital from 1996 December to 2008 December were retrospectively analyzed. RESULTS: There were 22 patients in primary group, and 22 in thyrotoxicosis group. Identical clinical features of both the groups: (1)It had a predilection in young men. (2)Main symptoms were limb movement disorder and fatigue, and paralysis recurrent attacked in most patients. (3) 40.9% to 68.2% patients had obvious incentives, and the common ones were a heavy meal, sweet drinks, or strenuous exercise. (4) Serum potassium levels of the two groups were obviously lower than the normal range. (5)In 20% patients of primary group and 25% patients of thyrotoxicosis secondary group, CK levels were higher than normal, while LDH and HBDH levels were normal. (6)The doses of potassium replishment were not correlated to serum potassium levels at the onset. Different clinical features of the two groups: (1) Patients of thyrotoxicosis group had hypermetabolism symptoms and thyroid dysfunction. Patients of primary group had no hypermetabolism symptoms, and all of them were euthyroid. (2)Serum potassium levels of thyrotoxicosis secondary group were lower than those of primary group significantly [(2.25 +/- 0.67) vs (2.78 +/- 0.49) mmol/L, P=0.007]. (3) Hyperkalemia is easier than primary group to rebound in thyrotoxicosis secondary group, after replenishment of potassium. CONCLUSION: Hypokalemic periodic paralysis has its clinical features, and patients with early diagnosis and replenishment of potassium in time have good prognosis. The doses of potassium replenishment are not determined by serum potassium levels at the onset. Hyperkalemia is easier to rebound in thyrotoxicosis secondary group after replenishment of potassium, serum potassium levels should be monitored closely, and hyperthyrosis radically cured. PMID- 20019781 TI - [Optimization of riboflavin sodium phosphate loading to calcium alginate floating microspheres by response surface methodology]. AB - OBJECTIVE: To investigate the preparation, optimization and in vitro properties of riboflavin sodium phosphate floating microspheres. METHODS: The floating microspheres composed of riboflavin sodium phosphate and calcium alginate were prepared using ion gelatin-oven drying method. RESULTS: The properties of the microspheres were investigated, including the buoyancy, release, appearance and entrapment efficiency. The formulation was optimized by response surface methodology (RSM). CONCLUSION: The optimized microspheres were round. The entrapment efficiency was 57.49%. All the microspheres could float on the artificial gastric juice over 8 hours. The release of the drug from the microspheres complied with Fick's diffusion. PMID- 20019782 TI - [Comparison and clinical significance of different imageological methods in the detection of transitional carcinoma of upper urinary tract: analysis of 234 cases]. AB - OBJECTIVE: To determine the diagnostic value of multislice CT urography (MSCTU) in patients with transitional cell carcinoma (TCC) of upper urinary tract by comparing other imageology methods used. METHODS: Two hundred and thirty four cases of transitional cell carcinoma of upper urinary tract, in which 82 cases were diagnosed pathologically with pelvic carcinoma and 152 cases with ureteral carcinoma, between June 2004 and September 2006 in our institute were enrolled in a retrospective study. Most of them underwent urological ultrasound, intravenous urogram (IVU), retrograde pyelography and MSCTU. We compared the positive rate (PR) and diagnostic rate (DR) of these methods used by chi-square test. RESULTS: Among the 234 cases, 215 patients underwent urologic ultrasound, in which 152 cases were detected to be abnormal, with the PR of 70.7%; Meanwhile, 58 cased were diagnosed by this examination, with the DR of 27.0%. IVU was performed in 193 patients and 132 cases were found to be abnormal, and the PR was 68.4%, 65 cases were diagnosed by IVU and the DR was 33.7%. And 132 patients underwent retrograde pyelography, by which 115 cases of lesion were detected, with the PR of 87.1%; In the meantime, 93 cases were diagnosed, with the DR of 70.5%. MSCTU was performed in 226 cases and 220 cases were found to be abnormal, and the PR was 97.3%; 214 cases were diagnosed by MSCTU, with the DR of 94.7%. The DR of detecting TCC of retrograde pyelography had statistically significant difference with that of ultrasound and IVU (P<0.001). As compared with retrograde pyelography, MSCTU had statistically significant superiority (P<0.001). CONCLUSION: To shorten the diagnosis time and mitigate the sufferings, patients with hematuria supposed to be TCC of upper urinary tract should be recommended to undergo MSCTU first. PMID- 20019783 TI - Increasing specificity of real time PCR to detect microRNA through primer design and annealing temperature increase. AB - OBJECTIVE: To investigate the non-specific and inaccurate amplification in cases of highly similar sequences among family members and the length heterogeneity of mature microRNA (miRNA), and find a condition that discriminates maximally among similar miRNA family members and detects the accurate expression level of miRNAs. METHODS: Primers with their mismatches and/or 3' end at different positions were designed. Amplification efficiencies were compared using matched and various mismatched primers by RNA-tailing and primer-extension RT-PCR at different annealing temperatures. Expression levels of several miRNAs in mouse brain were compared using miRNA specific primers with different termini. RESULTS: Raising annealing temperatures 12 degrees Celsius-14 degrees Celsius above the T(m) of the primers maximally increased amplification specificity without sacrificing sensitivity. Primers designed with their termini on or near variant positions could efficiently discriminate between miRNA isoforms. Using primers that terminated before the end of the mature miRNA did not miss the detection of shorter mature miRNA and provided accurate expression levels. CONCLUSION: Careful primer design and higher annealing temperature can increase specificity and accuracy of real time PCR miRNA detection. PMID- 20019784 TI - [Patterns of Enterococcus faecalis in infected root canals: an in vitro study]. AB - OBJECTIVE: To establish an in vitro root canal model infected by Enterococcus faecalis and to observe the morphology, distribution and relative position of Enterococcus faecalis in infected root canals. METHODS: Ten human healthy premolars extracted for orthodontic reasons were collected. Following sterilization, a total of 5 specimens were aseptically transferred to separate Eppendorf tubes containing 1.5 mL brain-heart infusion broth (BHI) inoculated with 0.1 mL Enterococcus faecalis suspension that had been adjusted to Mcfarland 5, and were incubated at 37 degrees Celsius for 21 days. The other 5 specimens were as controls. The roots of all specimens were then split into two halves along the mesiodistal axis. One half was processed with light microscopic (Brown & Brenn stain) to check the bacteria in dentinal tubules, and the other was observed with SEM to investigate the bacterial status in infected root canals. RESULTS: Enterococcus faecalis could penetrate into the dentinal tubules about 330-1 000 mum. A dense bacterial aggregation composed of Enterococcus faecalis and amorphous matrix was observed in the apical third of the root canals, whereas Enterococcus faecalis were seen free-floating or planktonic in the crown and middle third of the root canals . No microorganisms were found in the root canals of the controls. CONCLUSION: Enterococcus faecalis could form bacterial biofilm on the root canal walls and penetrate into the dentinal tubules. The in vitro model designed was simple, and had good practicability to make a further comparative evaluation of various antimicrobial methods in the reduction of intracanal bacteria. PMID- 20019785 TI - [Comparison of the models of acute hypoxia and hypoxic-ischemia in newborn piglets]. AB - OBJECTIVE: To compare the newborn piglet models of hypoxic ischemic brain damage in hypoxia and hypoxia combined with occlusion of both carotid arteries. METHODS: Twenty four 7-day-old piglets were divided into two groups. Group H: mechanical ventilation with low concentration of oxygen, Group HI: mechanical ventilation with low concentration of oxygen combined with occlusion of both carotid arteries. The piglets were inhaled with 10%, 8%, and 6% low-concentration oxygen for 30 min, and grouped into mild, moderate, and severe hypoxia ones. The changes of physiological parameter, cerebral blood flow and cerebral oxygen perfusion were detected. RESULTS: There were no significant differences in blood gas analysis of oxygen saturation, blood lactic acid and pH between the two groups(P>0.05).The mean arterial pressure of severe hypoxia in HI was significantly lower than in H (P<0.05). The cerebral blood flow in H and HI was relatively stable after different degrees of hypoxia. As compared with the cerebral blood flow perfusion in group H and HI, there were no significant differences between them(P>0.05). The cerebral oxygen perfusion in H and HI was significantly descent after different degrees of hypoxia(P<0.05). As compared with the cerebral oxygen perfusion in groups H and HI, there were no significant differences between them. CONCLUSION: H and HI have the same effect on physiological parameter, cerebral volume and cerebral oxygen perfusion of newborn piglets. The mechanical ventilation with low concentration of oxygen to newborn piglets can develop the HIBD model, it is not necessary to occlude carotid arteries. PMID- 20019786 TI - [Application of covered stent in the treatment of radiation-induced common carotid artery bleeding to a patient with nasoparyngeal carcinoma: a case report]. AB - Bleeding is a rare but fatal complication after radiotherapy for nasoparyngeal carcinoma (NPC). In this article, we report an NPC case treated with radiotherapy complicated with bleeding of the common carotid artery(CCA). A 44-year-old man with NPC was treated with external radiotherapy 19 years ago, and a second course of treatment to the same field was administrated because of local nasoparyngeal recurrence 4 years ago. The patient was admitted to our hospital for massive bleeding. Conservative therapy including routine medication, blood transfusion and local compression showed almost no effect. Emergency carotid angiography revealed bleeding from the left CCA, and a self-expanding covered stent was deployed through percutaneous transluminal femoral approach. After the placement of the covered stent, angiography showed the bleeding was successfully controlled. So self-expanding covered stenting may be a safe, effective and minimal-invasive option for bleeding of the carotid artery. Long-term follow-up is required for further evaluation. PMID- 20019787 TI - [Pregnancy in renal transplant recipient: a case report and review]. AB - In recent years, successful pregnancies in renal transplant women have been reported worldwide. However, pregnancy in renal transplant recipient is relatively rare in China. This paper reported a case of pregnancy 2 years post renal transplant. At the end of 28 weeks' gestation she had abnormal liver and renal function complicated with severe preeclampsia. Literatures were reviewed on considerations about pregnancy in renal transplant women. Maternal and neonatal outcomes can be improved by intensive care during pregnancy, proper immunosupression and timely termination of pregnancy. PMID- 20019788 TI - [Research progress of Rett syndrome causing gene MECP2--the structure, function and modulation of MECP2]. AB - Rett syndrome (RTT), an X-linked dominant neurodevelopmental disorder characterized by regression of language, stereotype hand movement and loss of purposeful hand use, is primarily caused by mutation of menthyl-CpG-binding protein 2 (MECP2). The 76 kb human MECP2 is characterized by three salient features: a very large intron 2 (60 kb), an 8.5 kb 3'-UTR with highly conserved regions and different polyadenylation sites, and a 40 kb intergenic region separating MECP2 from the nearest upstream gene. There are two isoforms of MeCP2, MeCP2e1 and MeCP2e2. The differences between the two isoforms, the function of the 3'-UTR and the long-range cis-regulatory sequences in the intergenic region were extensively studied. In contrast to initial report, recent studies show that MeCP2 binds not only to methylated promoters and silence transcription, but also to the sites outside of genes containing only a few of CpG islands. Furthermore, MeCP2 can function as both an activator and a repressor of transcription. PMID- 20019789 TI - [Diffusion spectrum magnetic resonance imaging]. AB - Diffusion spectrum imaging (DSI), a newly developed MRI technique, affords the capacity to map complex fiber architectures in tissues with sufficient angular resolution by imaging the spectra of tissue water diffusion. By contrast, diffusion tensor imaging (DTI), the currently widely used technique based on the 2nd order tensor model, obtains an approximation of the complex diffusion, and provides only one global maximal direction corresponding to the primary eigenvector for each voxel. As a generalized model-free diffusion imaging technique, firstly, DSI employs the probability density function to describe the diffusion process in each voxel; secondly, a sufficient dense signal sample derived from repeated applications of diffusion-weighed gradients ensures its capability to resolve the diffusion probability density function; thirdly, specific computer visualization techniques are used to extract the diffusion information and reconstruct the geometrical properties of tissue microstructure. The capacity to unravel complex tissue architecture, recent improvements in hardware and ongoing optimization of sequence design and algorithm enable a rapid growth of DSI for research use and future incorporation into clinical protocols. This paper introduces the basic principles of DSI and then compares the characteristics of DSI and DTI schemes. Finally, the typical applications of DSI to date are reviewed. PMID- 20019790 TI - Alu and b1 repeats have been selectively retained in the upstream and intronic regions of genes of specific functional classes. AB - Alu and B1 repeats are mobile elements that originated in an initial duplication of the 7SL RNA gene prior to the primate-rodent split about 80 million years ago and currently account for a substantial fraction of the human and mouse genome, respectively. Following the primate-rodent split, Alu and B1 elements spread independently in each of the two genomes in a seemingly random manner, and, according to the prevailing hypothesis, negative selection shaped their final distribution in each genome by forcing the selective loss of certain Alu and B1 copies. In this paper, contrary to the prevailing hypothesis, we present evidence that Alu and B1 elements have been selectively retained in the upstream and intronic regions of genes belonging to specific functional classes. At the same time, we found no evidence for selective loss of these elements in any functional class. A subset of the functional links we discovered corresponds to functions where Alu involvement has actually been experimentally validated, whereas the majority of the functional links we report are novel. Finally, the unexpected finding that Alu and B1 elements show similar biases in their distribution across functional classes, despite having spread independently in their respective genomes, further supports our claim that the extant instances of Alu and B1 elements are the result of positive selection. PMID- 20019791 TI - Predicting functional alternative splicing by measuring RNA selection pressure from multigenome alignments. AB - High-throughput methods such as EST sequencing, microarrays and deep sequencing have identified large numbers of alternative splicing (AS) events, but studies have shown that only a subset of these may be functional. Here we report a sensitive bioinformatics approach that identifies exons with evidence of a strong RNA selection pressure ratio (RSPR)--i.e., evolutionary selection against mutations that change only the mRNA sequence while leaving the protein sequence unchanged--measured across an entire evolutionary family, which greatly amplifies its predictive power. Using the UCSC 28 vertebrate genome alignment, this approach correctly predicted half to three-quarters of AS exons that are known binding targets of the NOVA splicing regulatory factor, and predicted 345 strongly selected alternative splicing events in human, and 262 in mouse. These predictions were strongly validated by several experimental criteria of functional AS such as independent detection of the same AS event in other species, reading frame-preservation, and experimental evidence of tissue-specific regulation: 75% (15/20) of a sample of high-RSPR exons displayed tissue specific regulation in a panel of ten tissues, vs. only 20% (4/20) among a sample of low RSPR exons. These data suggest that RSPR can identify exons with functionally important splicing regulation, and provides biologists with a dataset of over 600 such exons. We present several case studies, including both well-studied examples (GRIN1) and novel examples (EXOC7). These data also show that RSPR strongly outperforms other approaches such as standard sequence conservation (which fails to distinguish amino acid selection pressure from RNA selection pressure), or pairwise genome comparison (which lacks adequate statistical power for predicting individual exons). PMID- 20019792 TI - Dissecting early differentially expressed genes in a mixture of differentiating embryonic stem cells. AB - The differentiation of embryonic stem cells is initiated by a gradual loss of pluripotency-associated transcripts and induction of differentiation genes. Accordingly, the detection of differentially expressed genes at the early stages of differentiation could assist the identification of the causal genes that either promote or inhibit differentiation. The previous methods of identifying differentially expressed genes by comparing different cell types would inevitably include a large portion of genes that respond to, rather than regulate, the differentiation process. We demonstrate through the use of biological replicates and a novel statistical approach that the gene expression data obtained without prior separation of cell types are informative for detecting differentially expressed genes at the early stages of differentiation. Applying the proposed method to analyze the differentiation of murine embryonic stem cells, we identified and then experimentally verified Smarcad1 as a novel regulator of pluripotency and self-renewal. We formalized this statistical approach as a statistical test that is generally applicable to analyze other differentiation processes. PMID- 20019793 TI - Fungicide-driven evolution and molecular basis of multidrug resistance in field populations of the grey mould fungus Botrytis cinerea. AB - The grey mould fungus Botrytis cinerea causes losses of commercially important fruits, vegetables and ornamentals worldwide. Fungicide treatments are effective for disease control, but bear the risk of resistance development. The major resistance mechanism in fungi is target protein modification resulting in reduced drug binding. Multiple drug resistance (MDR) caused by increased efflux activity is common in human pathogenic microbes, but rarely described for plant pathogens. Annual monitoring for fungicide resistance in field isolates from fungicide treated vineyards in France and Germany revealed a rapidly increasing appearance of B. cinerea field populations with three distinct MDR phenotypes. All MDR strains showed increased fungicide efflux activity and overexpression of efflux transporter genes. Similar to clinical MDR isolates of Candida yeasts that are due to transcription factor mutations, all MDR1 strains were shown to harbor activating mutations in a transcription factor (Mrr1) that controls the gene encoding ABC transporter AtrB. MDR2 strains had undergone a unique rearrangement in the promoter region of the major facilitator superfamily transporter gene mfsM2, induced by insertion of a retrotransposon-derived sequence. MDR2 strains carrying the same rearranged mfsM2 allele have probably migrated from French to German wine-growing regions. The roles of atrB, mrr1 and mfsM2 were proven by the phenotypes of knock-out and overexpression mutants. As confirmed by sexual crosses, combinations of mrr1 and mfsM2 mutations lead to MDR3 strains with higher broad-spectrum resistance. An MDR3 strain was shown in field experiments to be selected against sensitive strains by fungicide treatments. Our data document for the first time the rising prevalence, spread and molecular basis of MDR populations in a major plant pathogen in agricultural environments. These populations will increase the risk of grey mould rot and hamper the effectiveness of current strategies for fungicide resistance management. PMID- 20019794 TI - Use of zebrafish to probe the divergent virulence potentials and toxin requirements of extraintestinal pathogenic Escherichia coli. AB - Extraintestinal pathogenic E. coli (ExPEC) cause an array of diseases, including sepsis, neonatal meningitis, and urinary tract infections. Many putative virulence factors that might modulate ExPEC pathogenesis have been identified through sequencing efforts, epidemiology, and gene expression profiling, but few of these genes have been assigned clearly defined functional roles during infection. Using zebrafish embryos as surrogate hosts, we have developed a model system with the ability to resolve diverse virulence phenotypes and niche specific restrictions among closely related ExPEC isolates during either localized or systemic infections. In side-by-side comparisons of prototypic ExPEC isolates, we observed an unexpectedly high degree of phenotypic diversity that is not readily apparent using more traditional animal hosts. In particular, the capacity of different ExPEC isolates to persist and multiply within the zebrafish host and cause disease was shown to be variably dependent upon two secreted toxins, alpha-hemolysin and cytotoxic necrotizing factor. Both of these toxins appear to function primarily in the neutralization of phagocytes, which are recruited in high numbers to sites of infection where they act as an essential host defense against ExPEC as well as less virulent E. coli strains. These results establish zebrafish as a valuable tool for the elucidation and functional analysis of both ExPEC virulence factors and host defense mechanisms. PMID- 20019795 TI - Evolutionary plasticity and innovations in complex metabolic reaction networks. AB - Genome-scale metabolic networks are highly robust to the elimination of enzyme coding genes. Their structure can evolve rapidly through mutations that eliminate such genes and through horizontal gene transfer that adds new enzyme-coding genes. Using flux balance analysis we study a vast space of metabolic network genotypes and their relationship to metabolic phenotypes, the ability to sustain life in an environment defined by an available spectrum of carbon sources. Two such networks typically differ in most of their reactions and have few essential reactions in common. Our observations suggest that the robustness of the Escherichia coli metabolic network to mutations is typical of networks with the same phenotype. We also demonstrate that networks with the same phenotype form large sets that can be traversed through single mutations, and that single mutations of different genotypes with the same phenotype can yield very different novel phenotypes. This means that the evolutionary plasticity and robustness of metabolic networks facilitates the evolution of new metabolic abilities. Our approach has broad implications for the evolution of metabolic networks, for our understanding of mutational robustness, for the design of antimetabolic drugs, and for metabolic engineering. PMID- 20019796 TI - Mobile antibiotic resistance encoding elements promote their own diversity. AB - Integrating conjugative elements (ICEs) are a class of bacterial mobile genetic elements that disseminate via conjugation and then integrate into the host cell genome. The SXT/R391 family of ICEs consists of more than 30 different elements that all share the same integration site in the host chromosome but often encode distinct properties. These elements contribute to the spread of antibiotic resistance genes in several gram-negative bacteria including Vibrio cholerae, the agent of cholera. Here, using comparative analyses of the genomes of several SXT/R391 ICEs, we found evidence that the genomes of these elements have been shaped by inter-ICE recombination. We developed a high throughput semi quantitative method to explore the genetic determinants involved in hybrid ICE formation. Recombinant ICE formation proved to be relatively frequent, and to depend on host (recA) and ICE (s065 and s066) loci, which can independently and potentially cooperatively mediate hybrid ICE formation. s065 and s066, which are found in all SXT/R391 ICEs, are orthologues of the bacteriophage lambda Red recombination genes bet and exo, and the s065/s066 recombination system is the first Red-like recombination pathway to be described in a conjugative element. Neither ICE excision nor conjugative transfer proved to be essential for generation of hybrid ICEs. Instead conjugation facilitates the segregation of hybrids and could provide a means to select for functional recombinant ICEs containing novel combinations of genes conferring resistance to antibiotics. Thus, ICEs promote their own diversity and can yield novel mobile elements capable of disseminating new combinations of antibiotic resistance genes. PMID- 20019797 TI - Nano-motion dynamics are determined by surface-tethered selectin mechanokinetics and bond formation. AB - The interaction of proteins at cellular interfaces is critical for many biological processes, from intercellular signaling to cell adhesion. For example, the selectin family of adhesion receptors plays a critical role in trafficking during inflammation and immunosurveillance. Quantitative measurements of binding rates between surface-constrained proteins elicit insight into how molecular structural details and post-translational modifications contribute to function. However, nano-scale transport effects can obfuscate measurements in experimental assays. We constructed a biophysical simulation of the motion of a rigid microsphere coated with biomolecular adhesion receptors in shearing flow undergoing thermal motion. The simulation enabled in silico investigation of the effects of kinetic force dependence, molecular deformation, grouping adhesion receptors into clusters, surface-constrained bond formation, and nano-scale vertical transport on outputs that directly map to observable motions. Simulations recreated the jerky, discrete stop-and-go motions observed in P selectin/PSGL-1 microbead assays with physiologic ligand densities. Motion statistics tied detailed simulated motion data to experimentally reported quantities. New deductions about biomolecular function for P-selectin/PSGL-1 interactions were made. Distributing adhesive forces among P-selectin/PSGL-1 molecules closely grouped in clusters was necessary to achieve bond lifetimes observed in microbead assays. Initial, capturing bond formation effectively occurred across the entire molecular contour length. However, subsequent rebinding events were enhanced by the reduced separation distance following the initial capture. The result demonstrates that vertical transport can contribute to an enhancement in the apparent bond formation rate. A detailed analysis of in silico motions prompted the proposition of wobble autocorrelation as an indicator of two-dimensional function. Insight into two-dimensional bond formation gained from flow cell assays might therefore be important to understand processes involving extended cellular interactions, such as immunological synapse formation. A biologically informative in silico system was created with minimal, high-confidence inputs. Incorporating random effects in surface separation through thermal motion enabled new deductions of the effects of surface constrained biomolecular function. Important molecular information is embedded in the patterns and statistics of motion. PMID- 20019798 TI - DNA specificity determinants associate with distinct transcription factor functions. AB - To elucidate how genomic sequences build transcriptional control networks, we need to understand the connection between DNA sequence and transcription factor binding and function. Binding predictions based solely on consensus predictions are limited, because a single factor can use degenerate sequence motifs and because related transcription factors often prefer identical sequences. The ETS family transcription factor, ETS1, exemplifies these challenges. Unexpected, redundant occupancy of ETS1 and other ETS proteins is observed at promoters of housekeeping genes in T cells due to common sequence preferences and the presence of strong consensus motifs. However, ETS1 exhibits a specific function in T cell activation; thus, unique transcriptional targets are predicted. To uncover the sequence motifs that mediate specific functions of ETS1, a genome-wide approach, chromatin immunoprecipitation coupled with high-throughput sequencing (ChIP-seq), identified both promoter and enhancer binding events in Jurkat T cells. A comparison with DNase I sensitivity both validated the dataset and also improved accuracy. Redundant occupancy of ETS1 with the ETS protein GABPA occurred primarily in promoters of housekeeping genes, whereas ETS1 specific occupancy occurred in the enhancers of T cell-specific genes. Two routes to ETS1 specificity were identified: an intrinsic preference of ETS1 for a variant of the ETS family consensus sequence and the presence of a composite sequence that can support cooperative binding with a RUNX transcription factor. Genome-wide occupancy of RUNX factors corroborated the importance of this partnership. Furthermore, genome-wide occupancy of co-activator CBP indicated tight co localization with ETS1 at specific enhancers, but not redundant promoters. The distinct sequences associated with redundant versus specific ETS1 occupancy were predictive of promoter or enhancer location and the ontology of nearby genes. These findings demonstrate that diversity of DNA binding motifs may enable variable transcription factor function at different genomic sites. PMID- 20019799 TI - Long-range activation of systemic immunity through peptidoglycan diffusion in Drosophila. AB - The systemic immune response of Drosophila is known to be induced both by septic injury and by oral infection with certain bacteria, and is characterized by the secretion of antimicrobial peptides (AMPs) into the haemolymph. To investigate other possible routes of bacterial infection, we deposited Erwinia carotovora (Ecc15) on various sites of the cuticle and monitored the immune response via expression of the AMP gene Diptericin. A strong response was observed to deposition on the genital plate of males (up to 20% of a septic injury response), but not females. We show that the principal response to genital infection is systemic, but that some AMPs, particularly Defensin, are induced locally in the genital tract. At late time points we detected bacteria in the haemolymph of immune deficient Relish(E20) flies, indicating that the genital plate can be a route of entry for pathogens, and that the immune response protects flies against the progression of genital infection. The protective role of the immune response is further illustrated by our observation that Relish(E20) flies exhibit significant lethality in response to genital Ecc15 infections. We next show that a systemic immune response can be induced by deposition of the bacterial elicitor peptidoglycan (PGN), or its terminal monomer tracheal cytotoxin (TCT), on the genital plate. This immune response is downregulated by PGRP-LB and Pirk, known regulators of the Imd pathway, and can be suppressed by the overexpression of PGRP-LB in the haemolymph compartment. Finally, we provide strong evidence that TCT can activate a systemic response by crossing epithelia, by showing that radiolabelled TCT deposited on the genital plate can subsequently be detected in the haemolymph. Genital infection is thus an intriguing new model for studying the systemic immune response to local epithelial infections and a potential route of entry for naturally occurring pathogens of Drosophila. PMID- 20019800 TI - Power-law scaling in the brain surface electric potential. AB - Recent studies have identified broadband phenomena in the electric potentials produced by the brain. We report the finding of power-law scaling in these signals using subdural electrocorticographic recordings from the surface of human cortex. The power spectral density (PSD) of the electric potential has the power law form P(f ) approximately Af(-chi) from 80 to 500 Hz. This scaling index, chi = 4.0+/-0.1, is conserved across subjects, area in the cortex, and local neural activity levels. The shape of the PSD does not change with increases in local cortical activity, but the amplitude, A, increases. We observe a "knee" in the spectra at f(0) approximately 75 Hz, implying the existence of a characteristic time scale tau = (2pif(0))(-1) approximately 2 - 4ms. Below f(0), we explore two power-law forms of the PSD, and demonstrate that there are activity-related fluctuations in the amplitude of a power-law process lying beneath the alpha/beta rhythms. Finally, we illustrate through simulation how, small-scale, simplified neuronal models could lead to these power-law observations. This suggests a new paradigm of non-oscillatory "asynchronous," scale-free, changes in cortical potentials, corresponding to changes in mean population-averaged firing rate, to complement the prevalent "synchronous" rhythm-based paradigm. PMID- 20019801 TI - A multivalent and cross-protective vaccine strategy against arenaviruses associated with human disease. AB - Arenaviruses are the causative pathogens of severe hemorrhagic fever and aseptic meningitis in humans, for which no licensed vaccines are currently available. Pathogen heterogeneity within the Arenaviridae family poses a significant challenge for vaccine development. The main hypothesis we tested in the present study was whether it is possible to design a universal vaccine strategy capable of inducing simultaneous HLA-restricted CD8+ T cell responses against 7 pathogenic arenaviruses (including the lymphocytic choriomeningitis, Lassa, Guanarito, Junin, Machupo, Sabia, and Whitewater Arroyo viruses), either through the identification of widely conserved epitopes, or by the identification of a collection of epitopes derived from multiple arenavirus species. By inoculating HLA transgenic mice with a panel of recombinant vaccinia viruses (rVACVs) expressing the different arenavirus proteins, we identified 10 HLA-A02 and 10 HLA A03-restricted epitopes that are naturally processed in human antigen-presenting cells. For some of these epitopes we were able to demonstrate cross-reactive CD8+ T cell responses, further increasing the coverage afforded by the epitope set against each different arenavirus species. Importantly, we showed that immunization of HLA transgenic mice with an epitope cocktail generated simultaneous CD8+ T cell responses against all 7 arenaviruses, and protected mice against challenge with rVACVs expressing either Old or New World arenavirus glycoproteins. In conclusion, the set of identified epitopes allows broad, non ethnically biased coverage of all 7 viral species targeted by our studies. PMID- 20019802 TI - Alternative splicing events are a late feature of pathology in a mouse model of spinal muscular atrophy. AB - Spinal muscular atrophy is a severe motor neuron disease caused by inactivating mutations in the SMN1 gene leading to reduced levels of full-length functional SMN protein. SMN is a critical mediator of spliceosomal protein assembly, and complete loss or drastic reduction in protein leads to loss of cell viability. However, the reason for selective motor neuron degeneration when SMN is reduced to levels which are tolerated by all other cell types is not currently understood. Widespread splicing abnormalities have recently been reported at end stage in a mouse model of SMA, leading to the proposition that disruption of efficient splicing is the primary mechanism of motor neuron death. However, it remains unclear whether splicing abnormalities are present during early stages of the disease, which would be a requirement for a direct role in disease pathogenesis. We performed exon-array analysis of RNA from SMN deficient mouse spinal cord at 3 time points, pre-symptomatic (P1), early symptomatic (P7), and late-symptomatic (P13). Compared to littermate control mice, SMA mice showed a time-dependent increase in the number of exons showing differential expression, with minimal differences between genotypes at P1 and P7, but substantial variation in late-symptomatic (P13) mice. Gene ontology analysis revealed differences in pathways associated with neuronal development as well as cellular injury. Validation of selected targets by RT-PCR confirmed the array findings and was in keeping with a shift between physiologically occurring mRNA isoforms. We conclude that the majority of splicing changes occur late in SMA and may represent a secondary effect of cell injury, though we cannot rule out significant early changes in a small number of transcripts crucial to motor neuron survival. PMID- 20019803 TI - Model of transcriptional activation by MarA in Escherichia coli. AB - The AraC family transcription factor MarA activates approximately 40 genes (the marA/soxS/rob regulon) of the Escherichia coli chromosome resulting in different levels of resistance to a wide array of antibiotics and to superoxides. Activation of marA/soxS/rob regulon promoters occurs in a well-defined order with respect to the level of MarA; however, the order of activation does not parallel the strength of MarA binding to promoter sequences. To understand this lack of correspondence, we developed a computational model of transcriptional activation in which a transcription factor either increases or decreases RNA polymerase binding, and either accelerates or retards post-binding events associated with transcription initiation. We used the model to analyze data characterizing MarA regulation of promoter activity. The model clearly explains the lack of correspondence between the order of activation and the MarA-DNA affinity and indicates that the order of activation can only be predicted using information about the strength of the full MarA-polymerase-DNA interaction. The analysis further suggests that MarA can activate without increasing polymerase binding and that activation can even involve a decrease in polymerase binding, which is opposite to the textbook model of activation by recruitment. These findings are consistent with published chromatin immunoprecipitation assays of interactions between polymerase and the E. coli chromosome. We find that activation involving decreased polymerase binding yields lower latency in gene regulation and therefore might confer a competitive advantage to cells. Our model yields insights into requirements for predicting the order of activation of a regulon and enables us to suggest that activation might involve a decrease in polymerase binding which we expect to be an important theme of gene regulation in E. coli and beyond. PMID- 20019804 TI - Transcriptome and proteome exploration to model translation efficiency and protein stability in Lactococcus lactis. AB - This genome-scale study analysed the various parameters influencing protein levels in cells. To achieve this goal, the model bacterium Lactococcus lactis was grown at steady state in continuous cultures at different growth rates, and proteomic and transcriptomic data were thoroughly compared. Ratios of mRNA to protein were highly variable among proteins but also, for a given gene, between the different growth conditions. The modeling of cellular processes combined with a data fitting modeling approach allowed both translation efficiencies and degradation rates to be estimated for each protein in each growth condition. Estimated translational efficiencies and degradation rates strongly differed between proteins and were tested for their biological significance through statistical correlations with relevant parameters such as codon or amino acid bias. These efficiencies and degradation rates were not constant in all growth conditions and were inversely proportional to the growth rate, indicating a more efficient translation at low growth rate but an antagonistic higher rate of protein degradation. Estimated protein median half-lives ranged from 23 to 224 min, underlying the importance of protein degradation notably at low growth rates. The regulation of intracellular protein level was analysed through regulatory coefficient calculations, revealing a complex control depending on protein and growth conditions. The modeling approach enabled translational efficiencies and protein degradation rates to be estimated, two biological parameters extremely difficult to determine experimentally and generally lacking in bacteria. This method is generic and can now be extended to other environments and/or other micro-organisms. PMID- 20019806 TI - Calculating evolutionary dynamics in structured populations. AB - Evolution is shaping the world around us. At the core of every evolutionary process is a population of reproducing individuals. The outcome of an evolutionary process depends on population structure. Here we provide a general formula for calculating evolutionary dynamics in a wide class of structured populations. This class includes the recently introduced "games in phenotype space" and "evolutionary set theory." There can be local interactions for determining the relative fitness of individuals, but we require global updating, which means all individuals compete uniformly for reproduction. We study the competition of two strategies in the context of an evolutionary game and determine which strategy is favored in the limit of weak selection. We derive an intuitive formula for the structure coefficient, sigma, and provide a method for efficient numerical calculation. PMID- 20019805 TI - Meta-analysis of inter-species liver co-expression networks elucidates traits associated with common human diseases. AB - Co-expression networks are routinely used to study human diseases like obesity and diabetes. Systematic comparison of these networks between species has the potential to elucidate common mechanisms that are conserved between human and rodent species, as well as those that are species-specific characterizing evolutionary plasticity. We developed a semi-parametric meta-analysis approach for combining gene-gene co-expression relationships across expression profile datasets from multiple species. The simulation results showed that the semi parametric method is robust against noise. When applied to human, mouse, and rat liver co-expression networks, our method out-performed existing methods in identifying gene pairs with coherent biological functions. We identified a network conserved across species that highlighted cell-cell signaling, cell adhesion and sterol biosynthesis as main biological processes represented in genome-wide association study candidate gene sets for blood lipid levels. We further developed a heterogeneity statistic to test for network differences among multiple datasets, and demonstrated that genes with species-specific interactions tend to be under positive selection throughout evolution. Finally, we identified a human-specific sub-network regulated by RXRG, which has been validated to play a different role in hyperlipidemia and Type 2 diabetes between human and mouse. Taken together, our approach represents a novel step forward in integrating gene co-expression networks from multiple large scale datasets to leverage not only common information but also differences that are dataset-specific. PMID- 20019807 TI - Distributed dynamical computation in neural circuits with propagating coherent activity patterns. AB - Activity in neural circuits is spatiotemporally organized. Its spatial organization consists of multiple, localized coherent patterns, or patchy clusters. These patterns propagate across the circuits over time. This type of collective behavior has ubiquitously been observed, both in spontaneous activity and evoked responses; its function, however, has remained unclear. We construct a spatially extended, spiking neural circuit that generates emergent spatiotemporal activity patterns, thereby capturing some of the complexities of the patterns observed empirically. We elucidate what kind of fundamental function these patterns can serve by showing how they process information. As self-sustained objects, localized coherent patterns can signal information by propagating across the neural circuit. Computational operations occur when these emergent patterns interact, or collide with each other. The ongoing behaviors of these patterns naturally embody both distributed, parallel computation and cascaded logical operations. Such distributed computations enable the system to work in an inherently flexible and efficient way. Our work leads us to propose that propagating coherent activity patterns are the underlying primitives with which neural circuits carry out distributed dynamical computation. PMID- 20019808 TI - Adaptive gain modulation in V1 explains contextual modifications during bisection learning. AB - The neuronal processing of visual stimuli in primary visual cortex (V1) can be modified by perceptual training. Training in bisection discrimination, for instance, changes the contextual interactions in V1 elicited by parallel lines. Before training, two parallel lines inhibit their individual V1-responses. After bisection training, inhibition turns into non-symmetric excitation while performing the bisection task. Yet, the receptive field of the V1 neurons evaluated by a single line does not change during task performance. We present a model of recurrent processing in V1 where the neuronal gain can be modulated by a global attentional signal. Perceptual learning mainly consists in strengthening this attentional signal, leading to a more effective gain modulation. The model reproduces both the psychophysical results on bisection learning and the modified contextual interactions observed in V1 during task performance. It makes several predictions, for instance that imagery training should improve the performance, or that a slight stimulus wiggling can strongly affect the representation in V1 while performing the task. We conclude that strengthening a top-down induced gain increase can explain perceptual learning, and that this top-down signal can modify lateral interactions within V1, without significantly changing the classical receptive field of V1 neurons. PMID- 20019809 TI - Aging mice show a decreasing correlation of gene expression within genetic modules. AB - In this work we present a method for the differential analysis of gene co expression networks and apply this method to look for large-scale transcriptional changes in aging. We derived synonymous gene co-expression networks from AGEMAP expression data for 16-month-old and 24-month-old mice. We identified a number of functional gene groups that change co-expression with age. Among these changing groups we found a trend towards declining correlation with age. In particular, we identified a modular (as opposed to uniform) decline in general correlation with age. We identified potential transcriptional mechanisms that may aid in modular correlation decline. We found that computationally identified targets of the NF KappaB transcription factor decrease expression correlation with age. Finally, we found that genes that are prone to declining co-expression tend to be co-located on the chromosome. Our results conclude that there is a modular decline in co expression with age in mice. They also indicate that factors relating to both chromosome domains and specific transcription factors may contribute to the decline. PMID- 20019810 TI - Protein C inhibitor--a novel antimicrobial agent. AB - Protein C inhibitor (PCI) is a heparin-binding serine proteinase inhibitor belonging to the family of serpin proteins. Here we describe that PCI exerts broad antimicrobial activity against bacterial pathogens. This ability is mediated by the interaction of PCI with lipid membranes, which subsequently leads to their permeabilization. As shown by negative staining electron microscopy, treatment of Escherichia coli or Streptococcus pyogenes bacteria with PCI triggers membrane disruption followed by the efflux of bacterial cytosolic contents and bacterial killing. The antimicrobial activity of PCI is located to the heparin-binding site of the protein and a peptide spanning this region was found to mimic the antimicrobial activity of PCI, without causing lysis or membrane destruction of eukaryotic cells. Finally, we show that platelets can assemble PCI on their surface upon activation. As platelets are recruited to the site of a bacterial infection, these results may explain our finding that PCI levels are increased in tissue biopsies from patients suffering from necrotizing fasciitis caused by S. pyogenes. Taken together, our data describe a new function for PCI in innate immunity. PMID- 20019811 TI - Fruit-surface flavonoid accumulation in tomato is controlled by a SlMYB12 regulated transcriptional network. AB - The cuticle covering plants' aerial surfaces is a unique structure that plays a key role in organ development and protection against diverse stress conditions. A detailed analysis of the tomato colorless-peel y mutant was carried out in the framework of studying the outer surface of reproductive organs. The y mutant peel lacks the yellow flavonoid pigment naringenin chalcone, which has been suggested to influence the characteristics and function of the cuticular layer. Large-scale metabolic and transcript profiling revealed broad effects on both primary and secondary metabolism, related mostly to the biosynthesis of phenylpropanoids, particularly flavonoids. These were not restricted to the fruit or to a specific stage of its development and indicated that the y mutant phenotype is due to a mutation in a regulatory gene. Indeed, expression analyses specified three R2R3 MYB-type transcription factors that were significantly down-regulated in the y mutant fruit peel. One of these, SlMYB12, was mapped to the genomic region on tomato chromosome 1 previously shown to harbor the y mutation. Identification of an additional mutant allele that co-segregates with the colorless-peel trait, specific down-regulation of SlMYB12 and rescue of the y phenotype by overexpression of SlMYB12 on the mutant background, confirmed that a lesion in this regulator underlies the y phenotype. Hence, this work provides novel insight to the study of fleshy fruit cuticular structure and paves the way for the elucidation of the regulatory network that controls flavonoid accumulation in tomato fruit cuticle. PMID- 20019812 TI - The dystrophin complex controls bk channel localization and muscle activity in Caenorhabditis elegans. AB - Genetic defects in the dystrophin-associated protein complex (DAPC) are responsible for a variety of pathological conditions including muscular dystrophy, cardiomyopathy, and vasospasm. Conserved DAPC components from humans to Caenorhabditis elegans suggest a similar molecular function. C. elegans DAPC mutants exhibit a unique locomotory deficit resulting from prolonged muscle excitation and contraction. Here we show that the C. elegans DAPC is essential for proper localization of SLO-1, the large conductance, voltage-, and calcium dependent potassium (BK) channel, which conducts a major outward rectifying current in muscle under the normal physiological condition. Through analysis of mutants with the same phenotype as the DAPC mutants, we identified the novel islo 1 gene that encodes a protein with two predicted transmembrane domains. We demonstrate that ISLO-1 acts as a novel adapter molecule that links the DAPC to SLO-1 in muscle. We show that a defect in either the DAPC or ISLO-1 disrupts normal SLO-1 localization in muscle. Consistent with observations that SLO-1 requires a high calcium concentration for full activation, we find that SLO-1 is localized near L-type calcium channels in muscle, thereby providing a mechanism coupling calcium influx with the outward rectifying current. Our results indicate that the DAPC modulates muscle excitability by localizing the SLO-1 channel to calcium-rich regions of C. elegans muscle. PMID- 20019813 TI - T cell detection of a B-cell tropic virus infection: newly-synthesised versus mature viral proteins as antigen sources for CD4 and CD8 epitope display. AB - Viruses that naturally infect cells expressing both MHC I and MHC II molecules render themselves potentially visible to both CD8+ and CD4+ T cells through the de novo expression of viral antigens. Here we use one such pathogen, the B lymphotropic Epstein-Barr virus (EBV), to examine the kinetics of these processes in the virally-infected cell, comparing newly synthesised polypeptides versus the mature protein pool as viral antigen sources for MHC I- and MHC II-restricted presentation. EBV-transformed B cell lines were established in which the expression of two cognate EBV antigens, EBNA1 and EBNA3B, could be induced and then completely suppressed by doxycycline-regulation. These cells were used as targets for CD8+ and CD4+ T cell clones to a range of EBNA1 and EBNA3B epitopes. For both antigens, when synthesis was induced, CD8 epitope display rose quickly to near maximum within 24 h, well before steady state levels of mature protein had been reached, whereas CD4 epitope presentation was delayed by 36-48 h and rose only slowly thereafter. When antigen expression was suppressed, despite the persistence of mature protein, CD8 epitope display fell rapidly at rates similar to that seen for the MHC I/epitope half-life in peptide pulse-chase experiments. By contrast, CD4 epitope display persisted for many days and, following peptide stripping, recovered well on cells in the absence of new antigen synthesis. We infer that, in virally-infected MHC I/II-positive cells, newly-synthesised polypeptides are the dominant source of antigen feeding the MHC I pathway, whereas the MHC II pathway is fed by the mature protein pool. Hence, newly infected cells are rapidly visible only to the CD8 response; by contrast, latent infections, in which viral gene expression has been extinguished yet viral proteins persist, will remain visible to CD4+ T cells. PMID- 20019814 TI - BCA2/Rabring7 promotes tetherin-dependent HIV-1 restriction. AB - Host cell factors can either positively or negatively regulate the assembly and egress of HIV-1 particles from infected cells. Recent reports have identified a previously uncharacterized transmembrane protein, tetherin/CD317/BST-2, as a crucial host restriction factor that acts during a late budding step in HIV-1 replication by inhibiting viral particle release. Although tetherin has been shown to promote the retention of nascent viral particles on the host cell surface, the precise molecular mechanisms that occur during and after these tethering events remain largely unknown. We here report that a RING-type E3 ubiquitin ligase, BCA2 (Breast cancer-associated gene 2; also called Rabring7, ZNF364 or RNF115), is a novel tetherin-interacting host protein that facilitates the restriction of HIV-1 particle production in tetherin-positive cells. The expression of human BCA2 in "tetherin-positive" HeLa, but not in "tetherin negative" HOS cells, resulted in a strong restriction of HIV-1 particle production. Upon the expression of tetherin in HOS cells, BCA2 was capable of inhibiting viral particle production as in HeLa cells. The targeted depletion of endogenous BCA2 by RNA interference (RNAi) in HeLa cells reduced the intracellular accumulation of viral particles, which were nevertheless retained on the plasma membrane. BCA2 was also found to facilitate the internalization of HIV-1 virions into CD63(+) intracellular vesicles leading to their lysosomal degradation. These results indicate that BCA2 accelerates the internalization and degradation of viral particles following their tethering to the cell surface and is a co-factor or enhancer for the tetherin-dependent restriction of HIV-1 release from infected cells. PMID- 20019815 TI - Pre-exposure to moving form enhances static form sensitivity. AB - BACKGROUND: Motion-defined form can seem to persist briefly after motion ceases, before seeming to gradually disappear into the background. Here we investigate if this subjective persistence reflects a signal capable of improving objective measures of sensitivity to static form. METHODOLOGY/PRINCIPAL FINDINGS: We presented a sinusoidal modulation of luminance, masked by a background noise pattern. The sinusoidal luminance modulation was usually subjectively invisible when static, but visible when moving. We found that drifting then stopping the waveform resulted in a transient subjective persistence of the waveform in the static display. Observers' objective sensitivity to the position of the static waveform was also improved after viewing moving waveforms, compared to viewing static waveforms for a matched duration. This facilitation did not occur simply because movement provided more perspectives of the waveform, since performance following pre-exposure to scrambled animations did not match that following pre exposure to smooth motion. Observers did not simply remember waveform positions at motion offset, since removing the waveform before testing reduced performance. CONCLUSIONS/SIGNIFICANCE: Motion processing therefore interacts with subsequent static visual inputs in a way that can improve performance in objective sensitivity measures. We suggest that the brief subjective persistence of motion defined forms that can occur after motion offsets is a consequence of the decay of a static form signal that has been transiently enhanced by motion processing. PMID- 20019816 TI - Coordinated regulation of SIV replication and immune responses in the CNS. AB - Central nervous system (CNS) invasion during acute-stage HIV-infection has been demonstrated in a small number of individuals, but there is no evidence of neurological impairment at this stage and virus infection in brain appears to be controlled until late-stage disease. Using our reproducible SIV macaque model to examine the earliest stages of infection in the CNS, we identified immune responses that differentially regulate inflammation and virus replication in the brain compared to the peripheral blood and lymphoid tissues. SIV replication in brain macrophages and in brain of SIV-infected macaques was detected at 4 days post-inoculation (p.i.). This was accompanied by upregulation of innate immune responses, including IFNbeta, IFNbeta-induced gene MxA mRNA, and TNFalpha. Additionally, IL-10, the chemokine CCL2, and activation markers in macrophages, endothelial cells, and astrocytes were all increased in the brain at four days p.i. We observed synchronous control of virus replication, cytokine mRNA levels and inflammatory markers (MHC Class II, CD68 and GFAP) by 14 days p.i.; however, control failure was followed by development of CNS lesions in the brain. SIV infection was accompanied by induction of the dominant-negative isoform of C/EBPbeta, which regulates SIV, CCL2, and IL6 transcription, as well as inflammatory responses in macrophages and astrocytes. This synchronous response in the CNS is in part due to the effect of the C/EBPbeta on virus replication and cytokine expression in macrophage-lineage cells in contrast to CD4+ lymphocytes in peripheral blood and lymphoid tissues. Thus, we have identified a crucial period in the brain when virus replication and inflammation are controlled. As in HIV-infected individuals, though, this control is not sustained in the brain. Our results suggest that intervention with antiretroviral drugs or anti-inflammatory therapeutics with CNS penetration would sustain early control. These studies further suggest that interventions should target HIV-infected individuals with increased CCL2 levels or HIV RNA in the CNS. PMID- 20019817 TI - Investigating the role of mitochondrial haplogroups in genetic predisposition to meningococcal disease. AB - BACKGROUND AND AIMS: Meningococcal disease remains one of the most important infectious causes of death in industrialized countries. The highly diverse clinical presentation and prognosis of Neisseria meningitidis infections are the result of complex host genetics and environmental interactions. We investigated whether mitochondrial genetic background contributes to meningococcal disease (MD) susceptibility. METHODOLOGY/PRINCIPAL FINDINGS: Prospective controlled study was performed through a national research network on MD that includes 41 Spanish hospitals. Cases were 307 paediatric patients with confirmed MD, representing the largest series of MD patients analysed to date. Two independent sets of ethnicity matched control samples (CG1 [N = 917]), and CG2 [N = 616]) were used for comparison. Cases and controls underwent mtDNA haplotyping of a selected set of 25 mtDNA SNPs (mtSNPs), some of them defining major European branches of the mtDNA phylogeny. In addition, 34 ancestry informative markers (AIMs) were genotyped in cases and CG2 in order to monitor potential hidden population stratification. Samples of known African, Native American and European ancestry (N = 711) were used as classification sets for the determination of ancestral membership of our MD patients. A total of 39 individuals were eliminated from the main statistical analyses (including fourteen gypsies) on the basis of either non Spanish self-reported ancestry or the results of AIMs indicating a European membership lower than 95%. Association analysis of the remaining 268 cases against CG1 suggested an overrepresentation of the synonym mtSNP G11719A variant (Pearson's chi-square test; adjusted P-value = 0.0188; OR [95% CI] = 1.63 [1.22 2.18]). When cases were compared with CG2, the positive association could not be replicated. No positive association has been observed between haplogroup (hg) status of cases and CG1/CG2 and hg status of cases and several clinical variants. CONCLUSIONS: We did not find evidence of association between mtSNPs and mtDNA hgs with MD after carefully monitoring the confounding effect of population sub structure. MtDNA variability is particularly stratified in human populations owing to its low effective population size in comparison with autosomal markers and therefore, special care should be taken in the interpretation of seeming signals of positive associations in mtDNA case-control association studies. PMID- 20019819 TI - Adolescent access to healthcare. PMID- 20019820 TI - Canada's youth up in smoke: The decriminalization of cannabis. PMID- 20019818 TI - Global demethylation of rat chondrosarcoma cells after treatment with 5-aza-2' deoxycytidine results in increased tumorigenicity. AB - Abnormal patterns of DNA methylation are observed in several types of human cancer. While localized DNA methylation of CpG islands has been associated with gene silencing, the effect that genome-wide loss of methylation has on tumorigenesis is not completely known. To examine its effect on tumorigenesis, we induced DNA demethylation in a rat model of human chondrosarcoma using 5-aza-2 deoxycytidine. Rat specific pyrosequencing assays were utilized to assess the methylation levels in both LINEs and satellite DNA sequences following 5-aza-2 deoxycytidine treatment. Loss of DNA methylation was accompanied by an increase in invasiveness of the rat chondrosarcoma cells, in vitro, as well as by an increase in tumor growth in vivo. Subsequent microarray analysis provided insight into the gene expression changes that result from 5-aza-2-deoxycytidine induced DNA demethylation. In particular, two genes that may function in tumorigenesis, sox-2 and midkine, were expressed at low levels in control cells but upon 5-aza-2 deoxycytidine treatment these genes became overexpressed. Promoter region DNA analysis revealed that these genes were methylated in control cells but became demethylated following 5-aza-2-deoxycytidine treatment. Following withdrawal of 5 aza-2-deoxycytidine, the rat chondrosarcoma cells reestablished global DNA methylation levels that were comparable to that of control cells. Concurrently, invasiveness of the rat chondrosarcoma cells, in vitro, decreased to a level indistinguishable to that of control cells. Taken together these experiments demonstrate that global DNA hypomethylation induced by 5-aza-2-deoxycytidine may promote specific aspects of tumorigenesis in rat chondrosarcoma cells. PMID- 20019821 TI - An approach to interviewing adolescents. PMID- 20019822 TI - What is new in paediatric epilepsy? AB - There are many recent advances in the understanding and management of epilepsy in children. Epidemiological data support the requirement for two or more unprovoked seizures for the diagnosis. New epilepsy syndromes abound as magnetic resonance imaging has uncovered new causative brain malformations with important implications for counseling and treatment. Molecular genetics has uncovered the basic mechanisms for several epilepsy syndromes. Indications for new drugs are slowly being defined, while nonmedication treatments for intractable patients have been further developed (ketogenic diet, vagus nerve stimulation, cortical resection). Clinical studies show that the chance of long-term remission can be predicted fairly accurately for many children. Death from childhood epilepsy is exceedingly rare. Despite these gains, the social outcome in adulthood is often unsatisfactory. PMID- 20019823 TI - The paediatrician and cardiac auscultation. AB - The cardiac auscultation (CA) skills of paediatric residents and office-based paediatricians have recently been shown to be suboptimal. CA is known to have a high degree of specificity and sensitivity, and is inexpensive. New teaching aids and availability of surrogate patient heart sounds and murmurs now allow most physicians to acquire CA skills. These teaching aids should be available in all medical schools and in all postgraduate paediatric training programs. While the relationship between musicality and CA skill has not been proven, the author assumes this relationship to be valid. Specific learning objectives in CA should be established. Recognizing that the current trend is away from clinical examinations, students frequently are unaware that a CA learning deficit exists. Therefore, students' CA skills should be evaluated before medical school graduation and at the Royal College of Physicians and Surgeons specialty examination. Students with amusia (inability to distinguish pitch of sound) should be identified and consideration should be given to directing them away from a clinical specialty in which CA is important. Further study is required in the physiology of learning of CA. Appropriate action by medical school and paediatric postgraduate program directors and the Royal College will effect a higher standard of patient care by increasing CA skills, resulting in a more financially efficient system - something everyone is trying to achieve in these times of increasing medical care costs. PMID- 20019824 TI - The unrealistic quest to thinness. PMID- 20019825 TI - The lateral chest radiograph. PMID- 20019826 TI - Should the lateral chest radiograph be routine in the diagnosis of pneumonia in children? A review of the literature. AB - OBJECTIVE: To assess the evidence of the additive diagnostic value of the lateral chest radiograph to the frontal chest radiograph in children (zero to 18 years) with suspected pneumonia in the emergency department. METHODS: The MEDLINE database (1966 to 2001) was searched to identify the highest level of evidence available in support of the lateral. The key words used included chest radiograph, pneumonia and chest x-ray. RESULTS: Only four controlled studies were identified that assessed the role of the lateral chest x-ray. Three of these studies were retrospective. In the first study, the addition of the lateral to the frontal chest x-ray in 414 children would have detected an additional nine patients (2.3%) with pneumonia. In the second study, involving 179 patients, a high level of agreement was found between the interpretations based on the frontal chest x-ray alone and those based on both views. A third study of 359 children assessed the criterion validity of a frontal chest x-ray alone. Based upon a radiologist's interpretation of both views as the gold standard, the sensitivity of the frontal view alone was 86%. A fourth study of 158 children with suspected pneumonia revealed that the frontal view alone was diagnostic of pneumonia in 106 of 109 (97%) patients. CONCLUSION: There is no randomized controlled trial evidence to support the additive value of the lateral to the frontal chest x-ray in the diagnosis of children with pneumonia. Further prospective studies are required to determine if the addition of the lateral chest x-ray will modify therapy, prevent complications or whether it is cost efficient. PMID- 20019827 TI - In children with idiopathic short stature, what advantage does administering recombinant growth hormone have over observation in final adult height?: Part A: Evidence-based answer and summary. PMID- 20019828 TI - In children with idiopathic short stature, what advantage does administering recombinant growth hormone have over observation in final adult height?: Part B: Clinical commentary. PMID- 20019829 TI - Case 1: The girl who couldn't sit still. PMID- 20019831 TI - Age limits and adolescents. PMID- 20019830 TI - Case 2: First do no harm. PMID- 20019832 TI - Smoking and teens: What you should know. PMID- 20019834 TI - Stromal cells and integrins: conforming to the needs of the tumor microenvironment. AB - The microenvironment of a tumor is constituted of a heterogenous population of stromal cells, extracellular matrix components, and secreted factors, all of which make the tumor microenvironment distinct from that of normal tissue. Unlike healthy cells, tumor cells require these unique surroundings to metastasize, spread, and form a secondary tumor at a distant site. In this review, we discuss that stromal cells such as fibroblasts and immune cells including macrophages, their secreted factors, such as vascular endothelial growth factor, transforming growth factor beta, and various chemokines, and the integrins that connect the various cell types play a particularly vital role in the survival of a growing tumor mass. Macrophages and fibroblasts are uniquely plastic cells because they are not only able to switch from tumor suppressing to tumor supporting phenotypes but also able to adopt various tumor-supporting functions based on their location within the microenvironment. Integrins serve as the backbone for all of these prometastatic operations because their function as cell-cell and cell-matrix signal transducers are important for the heterogenous components of the microenvironment to communicate. PMID- 20019835 TI - Human immunodeficiency virus type 1 Tat accelerates Kaposi sarcoma-associated herpesvirus Kaposin A-mediated tumorigenesis of transformed fibroblasts in vitro as well as in nude and immunocompetent mice. AB - Kaposi sarcoma-associated herpesvirus (KSHV) is necessary but not sufficient to cause Kaposi sarcoma (KS). Coinfection with human immunodeficiency virus type 1 (HIV-1), in the absence of antiretroviral suppressive therapy, drastically increases the risk of KS. Previously, we identified that HIV-1 transactivative transcription protein (Tat) was an important cofactor that activated lytic cycle replication of KSHV. Here, we further investigated the potential of Tat to influence tumorigenesis induced by KSHV Kaposin A, a product of KSHV that was encoded by the open reading frame K12 (a KSHV-transforming gene). By using colony formation in soft agar, (3)H-TdR incorporation, cell cycle, and microarray gene expression analyses, we demonstrated that Tat enhanced proliferation as well as mitogen-activated protein kinase, signal transducer and activator of transcription 3, and phosphatidylinositol 3-kinase/protein kinase B signaling induced by Kaposin A in NIH3T3 cells. Animal experiments further demonstrated that Tat accelerated tumorigenesis by Kaposin A in athymic nu/nu mice. Cells obtained from primary tumors of nude mice succeeded inducing tumors in immunocompetent mice. These data suggest that Tat can accelerate tumorigenesis induced by Kaposin A. Our data present the first line of evidence that Tat may participate in KS pathogenesis by collaborating with Kaposin A in acquired immunodeficiency syndrome (AIDS)-related KS (AIDS-KS) patients. Our data also suggest that the model for Kaposin and Tat-mediated oncogenesis will contribute to our understanding of the pathogenesis of AIDS-KS at the molecular level and may even be important in exploring a novel therapeutic method for AIDS-KS. PMID- 20019836 TI - Peptides derived from type IV collagen, CXC chemokines, and thrombospondin-1 domain-containing proteins inhibit neovascularization and suppress tumor growth in MDA-MB-231 breast cancer xenografts. AB - Angiogenesis or neovascularization, the process of new blood vessel formation from preexisting microvasculature, involves interactions among several cell types including parenchymal, endothelial cells, and immune cells. The formation of new vessels is tightly regulated by a balance between endogenous proangiogenic and antiangiogenic factors to maintain homeostasis in tissue; tumor progression and metastasis in breast cancer have been shown to be angiogenesis-dependent. We previously introduced a systematic methodology to identify putative endogenous antiangiogenic peptides and validated these predictions in vitro in human umbilical vein endothelial cell proliferation and migration assays. These peptides are derived from several protein families including type IV collagen, CXC chemokines, and thrombospondin-1 domain-containing proteins. On the basis of the results from the in vitro screening, we have evaluated the ability of one peptide selected from each family named pentastatin-1, chemokinostatin-1, and properdistatin, respectively, to suppress angiogenesis in an MDA-MB-231 human breast cancer orthotopic xenograft model in severe combined immunodeficient mice. Peptides were administered intraperitoneally once per day. We have demonstrated significant suppression of tumor growth in vivo and subsequent reductions in microvascular density, indicating the potential of these peptides as therapeutic agents for breast cancer. PMID- 20019837 TI - Co-overexpression of Met and hepatocyte growth factor promotes systemic metastasis in NCI-H460 non-small cell lung carcinoma cells. AB - Complete resection of early-stage non-small cell lung cancer (NSCLC) is potentially curative, yet approximately 50% of patients are at risk for developing metastatic recurrence. Met, the receptor for hepatocyte growth factor (HGF) is a receptor tyrosine kinase with demonstrated roles in regulating cellular proliferation, motility, morphogenesis, and apoptosis. Met receptor and its ligand, HGF, are commonly overexpressed in NSCLC, and their overexpression has been associated with poor prognosis, which could potentially involve a paracrine and/or autocrine activation loop. However, there is as yet no direct evidence that HGF-Met signaling directly promotes metastasis in NSCLC cells. Using retroviral transduction, we overexpressed the human c-met and hgf complementary DNA, alone or in combination in the NCI-H460 human large cell carcinoma cell line. The HGF/Met co-overexpressing (H460-HGF/Met) cells demonstrated enhanced tumorigenicity in xenograft SCID mice. When these cells are implanted orthotopically into the lungs of nude rats, only the H460-HGF/Met cells showed higher spontaneous metastases to distant organs including bone, brain, and kidney. These results provide evidence that autocrine overactivation of the Met- HGF loop enhances systemic metastases in NSCLC. Targeted interference of this loop may potentially be an effective adjuvant therapy to improve survival of early-stage NSCLC patients. PMID- 20019833 TI - The War on Cancer rages on. AB - In 1971, the "War on Cancer" was launched by the US government to cure cancer by the 200-year anniversary of the founding of the United States of America, 1976. This article briefly looks back at the progress that has been made in cancer research and compares progress made in other areas of human affliction. While progress has indeed been made, the battle continues to rage on. PMID- 20019838 TI - Nuclear magnetic resonance detects phosphoinositide 3-kinase/Akt-independent traits common to pluripotent murine embryonic stem cells and their malignant counterparts. AB - Pluripotent embryonic stem (ES) cells, a potential source of somatic precursors for cell therapies, cause tumors after transplantation. Studies of mammalian carcinogenesis using nuclear magnetic resonance (NMR) spectroscopy have revealed changes in the choline region, particularly increased phosphocholine (PCho) content. High PCho levels in murine ES (mES) cells have recently been attributed to cell pluripotency. The phosphoinositide 3-kinase (PI3K)/Akt pathway has been implicated in tumor-like properties of mES cells. This study aimed to examine a potential link between the metabolic profile associated with choline metabolism of pluripotent mES cells and PI3K/Akt signaling. We used mES (ES-D3) and murine embryonal carcinoma cells (EC-F9) and compared the metabolic profiles of 1) pluripotent mES (ESD0), 2) differentiated mES (ESD14), and 3) pluripotent F9 cells. Involvement of the PI3K/Akt pathway was assessed using LY294002, a selective PI3K inhibitor. Metabolic profiles were characterized in the extracted polar fraction by (1)H NMR spectroscopy. Similarities were found between the levels of choline phospholipid metabolites (PCho/total choline and PCho/glycerophosphocholine [GPCho]) in ESD0 and F9 cell spectra and a greater than five-fold decrease of the PCho/GPCho ratio associated with mES cell differentiation. LY294002 caused no significant change in relative PCho levels but led to a greater-than two-fold increase in PCho/GPCho ratios. These results suggest that the PCho/GPCho ratio is a metabolic trait shared by pluripotent and malignant cells and that PI3K does not underlie its development. It is likely that the signature identified here in a mouse model may be relevant for safe therapeutic applications of human ES cells. PMID- 20019839 TI - CCL2 is a negative regulator of AMP-activated protein kinase to sustain mTOR complex-1 activation, survivin expression, and cell survival in human prostate cancer PC3 cells. AB - CCL2 is a cytokine prevalent in the prostate cancer tumor microenvironment. Recently, we reported that CCL2 induces the mammalian target of rapamycin (mTOR) pathway to promote prostate cancer PC3 cell survival; however, the mechanism used by CCL2 to maintain mTOR complex-1 (mTORC1) activation requires clarification. This study demonstrates that upon serum starvation, CCL2 functions as a negative regulator of AMP-activated protein kinase (AMPK) by decreasing phosphorylation at its major regulatory site (Thr(172)) in PC3, DU145, and C4-2B prostate cancer cells. The CCL2-mediated AMPK regulation decreased raptor phosphorylation (Ser(792)) resulting in hyperactivation of mTORC1. D942, a pharmacological activator of AMPK, stunted CCL2-induced mTORC1 activity, survivin expression, and cell survival without significantly affecting Akt activity. CCL2, however, conferred some resistance to the lethal effect of D942 compared with untreated cells. By using Akt-specific inhibitor X, it was shown that Akt inactivation did not cause an increase in AMPK phosphorylation in CCL2-stimulated cells, suggesting that CCL2-mediated negative regulation of AMPK is independent of Akt. Furthermore, bisindolylmaleimide-V, a specific inhibitor of p70(S6K), stunted survivin expression and induced cell death in CCL2-treated PC3. Altogether, these findings suggest that CCL2 hyperactivates mTORC1 through simultaneous regulation of both AMPK and Akt pathways and reveals a new network that promotes prostate cancer: CCL2-AMPK-mTORC1-survivin. PMID- 20019840 TI - Twist modulates breast cancer stem cells by transcriptional regulation of CD24 expression. AB - The cancer stem cell paradigm postulates that dysregulated tissue-specific stem cells or progenitor cells are precursors for cancer biogenesis. Consequently, identifying cancer stem cells is crucial to our understanding of cancer progression and for the development of novel therapeutic agents. In this study, we demonstrate that the overexpression of Twist in breast cells can promote the generation of a breast cancer stem cell phenotype characterized by the high expression of CD44, little or no expression of CD24, and increased aldehyde dehydrogenase 1 activity, independent of the epithelial-mesenchymal transition. In addition, Twist-overexpressing cells exhibit high efflux of Hoechst 33342 and Rhodamine 123 as a result of increased expression of ABCC1 (MRP1) transporters, a property of cancer stem cells. Moreover, we show that transient expression of Twist can induce the stem cell phenotype in multiple breast cell lines and that decreasing Twist expression by short hairpin RNA in Twist-overexpressing transgenic cell lines MCF-10A/Twist and MCF-7/Twist as well as in MDA-MB-231 partially reverses the stem cell molecular signature. Importantly, we show that inoculums of only 20 cells of the Twist-overexpressing CD44(+)/CD24(-/low) subpopulation are capable of forming tumors in the mammary fat pad of severe combined immunodeficient mice. Finally, with respect to mechanism, we provide data to indicate that Twist transcriptionally regulates CD24 expression in breast cancer cells. Taken together, our data demonstrate the direct involvement of Twist in generating a breast cancer stem cell phenotype through down-regulation of CD24 expression and independent of an epithelial-mesenchymal transition. PMID- 20019841 TI - Epigenetic silencing of CRABP2 and MX1 in head and neck tumors. AB - Head and neck squamous cell carcinoma (HNSCC) is a heterogeneous disease affecting the epithelium of the oral cavity, pharynx and larynx. Conditions of most patients are diagnosed at late stages of the disease, and no sensitive and specific predictors of aggressive behavior have been identified yet. Therefore, early detection and prognostic biomarkers are highly desirable for a more rational management of the disease. Hypermethylation of CpG islands is one of the most important epigenetic mechanisms that leads to gene silencing in tumors and has been extensively used for the identification of biomarkers. In this study, we combined rapid subtractive hybridization and microarray analysis in a hierarchical manner to select genes that are putatively reactivated by the demethylating agent 5-aza-2'-deoxycytidine (5Aza-dC) in HNSCC cell lines (FaDu, UM-SCC-14A, UM-SCC-17A, UM-SCC-38A). This combined analysis identified 78 genes, 35 of which were reactivated in at least 2 cell lines and harbored a CpG island at their 5' region. Reactivation of 3 of these 35 genes (CRABP2, MX1, and SLC15A3) was confirmed by quantitative real-time polymerase chain reaction (PCR; fold change, >or=3). Bisulfite sequencing of their CpG islands revealed that they are indeed differentially methylated in the HNSCC cell lines. Using methylation specific PCR, we detected a higher frequency of CRABP2 (58.1% for region 1) and MX1 (46.3%) hypermethylation in primary HNSCC when compared with lymphocytes from healthy individuals. Finally, absence of the CRABP2 protein was associated with decreased disease-free survival rates, supporting a potential use of CRABP2 expression as a prognostic biomarker for HNSCC patients. PMID- 20019842 TI - A regulatory polymorphism at position -309 in PTPRCAP is associated with susceptibility to diffuse-type gastric cancer and gene expression. AB - PTPRCAP (CD45-AP) is a positive regulator of protein tyrosine phosphatase PTPRC (CD45), which activates Src family kinases implicated in tumorigenesis. Single nucleotide polymorphism (SNP) rs869736 located at position -309 of the PTPRCAP promoter was associated with susceptibility to diffuse-type gastric cancer in the current case-control study. The minor-allele homozygote was significantly associated with a 2.5-fold increased susceptibility to diffuse-type gastric cancer (P = .0021, n = 252), but not to intestinal-type (P = .30, n = 178), versus the major-allele homozygote, when comparing unrelated Korean patients with healthy controls (n = 406). Nine other SNPs were in nearly perfect linkage disequilibrium (r(2) >or= 0.97) with this SNP, exhibiting the same association, and spread out for 26 kb on chromosome 11q13.1 covering RPS6KB2, PTPRCAP, CORO1B, and GPR152. Among the four genes, however, only PTPRCAP expression was affected by haplotypes of the 10 SNPs. Endogenous transcript levels of PTPRCAP were linearly correlated with copy numbers (0, 1, and 2) of the risk-haplotype (P = .0060) in 12 lymphoblastoid cells derived from blood samples, but those of the other three genes were not. Furthermore, the cancer-risk, minor-allele T of rs869736 increased both promoter activity and specific nuclear protein-binding affinity than the nonrisk, major-allele G in luciferase reporter and electrophoretic mobility shift assays, respectively. Accordingly, the minor allele of rs869736 in the PTPRCAP promoter is associated with increased susceptibility to diffuse-type gastric cancer by increasing PTPRCAP expression, possibly leading to activation of the oncogenic Src family kinases. PMID- 20019844 TI - Chemotherapeutic drug-induced ABCG2 promoter demethylation as a novel mechanism of acquired multidrug resistance. AB - ABCG2 is an efflux transporter conferring multidrug resistance (MDR) on cancer cells. However, the initial molecular events leading to its up-regulation in MDR tumor cells are poorly understood. Herein, we explored the impact of drug treatment on the methylation status of the ABCG2 promoter and consequent reactivation of ABCG2 gene expression in parental tumor cell lines and their MDR sublines. We demonstrate that ABCG2 promoter methylation is common in T-cell acute lymphoblastic leukemia (T-ALL) lines, also present in primary T-ALL lymphoblast specimens. Furthermore, drug selection with sulfasalazine and topotecan induced a complete demethylation of the ABCG2 promoter in the T-ALL and ovarian carcinoma model cell lines CCRF-CEM and IGROV1, respectively. This resulted in a dramatic induction of ABCG2 messenger RNA levels (235- and 743 fold, respectively) and consequent acquisition of an ABCG2-dependent MDR phenotype. Quantitative genomic polymerase chain reaction and ABCG2 promoter luciferase reporter assay did not reveal ABCG2 gene amplification or differential transcriptional trans-activation, which could account for ABCG2 up-regulation in these MDR cells. Remarkably, mimicking cytotoxic bolus drug treatment through 12- to 24-hour pulse exposure of ABCG2-silenced leukemia cells, to clinically relevant concentrations of the chemotherapeutic agents daunorubicin and mitoxantrone, resulted in a marked transcriptional up-regulation of ABCG2. Our findings establish that antitumor drug-induced epigenetic reactivation of ABCG2 gene expression in cancer cells is an early molecular event leading to MDR. These findings have important implications for the emergence, clonal selection, and expansion of malignant cells with the MDR phenotype during chemotherapy. PMID- 20019843 TI - Albumin-binding and tumor vasculature determine the antitumor effect of 15-deoxy Delta-(12,14)-prostaglandin-J(2) in vivo. AB - 15-Deoxy-Delta(12,14)-prostaglandin-J(2) (15d-PGJ(2)), a peroxisome proliferator activated receptor gamma (PPARgamma) agonist, induces cell death in tumor cells in vitro; however, no study showed its in vivo effect on tumors. Here, we report that 15d-PGJ(2) shows antitumor effects in vivo in mice. However, its effects correlate with tumor uptake of albumin, to which it reversibly binds. 15d-PGJ(2) induces cell death in B16F10 melanoma and C26 colon carcinoma cells in vitro. These effects were not elicited through PPARgamma-dependent pathways because an irreversible PPARgamma antagonist GW9662 did not inhibit these effects. Caspase- and nuclear factor kappaB- (NF-kappaB) dependent pathways were found to be involved as determined with caspase-3/7 fluorescent assay and NF-kappaB containing plasmid transfection assay, respectively. Noticeably, 15d-PGJ(2) had significantly stronger effects in C26 cells compared with B16 cells in all assays. However, in vivo, there was no effect on C26 tumors, yet it significantly inhibited the B16 tumor growth in mice by 75%. We found that 15d-PGJ(2) rapidly bound to albumin and in vivo albumin greatly distributed to B16 tumors compared with C26 tumors, shown with gamma-camera imaging and immunohistochemical staining. Albumin accumulation can be attributed to the large blood vessel diameter in B16 tumors and an enhanced permeability and retention effect. These findings suggest that 15d-PGJ(2) can be an effective therapeutic agent for cancer, although its effects seem to be limited to the tumors allowing albumin penetration. PMID- 20019846 TI - Improving seroreactivity-based detection of glioma. AB - Seroreactivity profiling emerges as valuable technique for minimal invasive cancer detection. Recently, we provided first evidence for the applicability of serum profiling of glioma using a limited number of immunogenic antigens. Here, we screened 57 glioma and 60 healthy sera for autoantibodies against 1827 Escherichia coli expressed clones, including 509 in-frame peptide sequences. By a linear support vector machine approach, we calculated mean specificity, sensitivity, and accuracy of 100 repetitive classifications. We were able to differentiate glioma sera from sera of the healthy controls with a specificity of 90.28%, a sensitivity of 87.31% and an accuracy of 88.84%. We were also able to differentiate World Health Organization grade IV glioma sera from healthy sera with a specificity of 98.45%, a sensitivity of 80.93%, and an accuracy of 92.88%. To rank the antigens according to their information content, we computed the area under the receiver operator characteristic curve value for each clone. Altogether, we found 46 immunogenic clones including 16 in-frame clones that were informative for the classification of glioma sera versus healthy sera. For the separation of glioblastoma versus healthy sera, we found 91 informative clones including 26 in-frame clones. The best-suited in-frame clone for the classification glioma sera versus healthy sera corresponded to the vimentin gene (VIM) that was previously associated with glioma. In the future, autoantibody signatures in glioma not only may prove useful for diagnosis but also offer the prospect for a personalized immune-based therapy. PMID- 20019847 TI - For coughs, colds and SARS, wear a mask. PMID- 20019848 TI - SARS and the maternal neonatal unit. PMID- 20019845 TI - Regulation of membrane-type 4 matrix metalloproteinase by SLUG contributes to hypoxia-mediated metastasis. AB - The hypoxic tumor environment has been shown to be critical to cancer metastasis through the promotion of angiogenesis, induction of epithelial-mesenchymal transition (EMT), and acquisition of invasive potential. However, the impact of hypoxia on the expression profile of the proteolytic enzymes involved in invasiveness is relatively unknown. Membrane-type 4 matrix metalloproteinase (MT4 MMP) is a glycosyl-phosphatidyl inositol-anchored protease that has been shown to be overexpressed in human cancers. However, detailed mechanisms regarding the regulation and function of MT4-MMP expression in tumor cells remain unknown. Here, we demonstrate that hypoxia or overexpression of hypoxia-inducible factor 1alpha (HIF-1alpha) induced MT4-MMP expression in human cancer cells. Activation of SLUG, a transcriptional factor regulating the EMT process of human cancers, by HIF-1alpha was critical for the induction of MT4-MMP under hypoxia. SLUG regulated the transcription of MT4-MMP through direct binding to the E-box located in its proximal promoter. Short-interference RNA-mediated knockdown of MT4-MMP attenuated in vitro invasiveness and in vivo pulmonary colonization of tumor cells without affecting cell migratory ability. MT4-MMP promoted invasiveness and pulmonary colonization through modulation of the expression profile of MMPs and angiogenic factors. Finally, coexpression of HIF-1alpha and MT4-MMP in human head and neck cancer was predictive of a worse clinical outcome. These findings establish a novel signaling pathway for hypoxia-mediated metastasis and elucidate the underlying regulatory mechanism and functional significance of MT4-MMP in cancer metastasis. PMID- 20019849 TI - The SARS outbreak and community paediatrics. PMID- 20019851 TI - Prophylaxis of respiratory syncytial virus in Canada in 2003. AB - Passive immunization of high-risk children with the humanized monoclonal antibody palivizumab is the mainstay of respiratory syncytial virus (RSV) prophylaxis in Canada in 2003. This product appears to be safe, and it prevents the majority of RSV hospitalizations in infants born before 36 weeks gestational age, and about half in children under 24 months of age with hemodynamically significant congenital heart disease. However, the high cost of palivizumab and the fact that at least 12 infants need to be treated throughout RSV season to prevent one hospitalization make it difficult to determine the ideal indications for the product. Because these high-risk infants account for a minority of RSV hospitalizations, it is desirable to search for a prophylactic strategy that is practical to apply in all infants. PMID- 20019850 TI - Respiratory syncytial virus vaccine: Is it coming? AB - Respiratory syncytial virus (RSV) is recognized as an important cause of childhood morbidity and mortality. Vaccine development has been challenging in young infants and has required the advent of molecular technologies to optimize the safety profile, while maintaining immunogenicity of live-attenuated vaccines. Protein-based vaccines have been evaluated in clinical trials and are promising candidates for RSV-primed populations or for maternal vaccination to provide early life protection. This review provides a summary of the need for an RSV vaccine, as well as the challenges and progress in the vaccine's development. PMID- 20019852 TI - Metapneumovirus and its place in childhood. AB - Human metapneumovirus (hMPV) was recently identified as a cause of acute upper and lower respiratory tract infection in children and adults. The epidemiology is similar to that exhibited by respiratory syncytial virus, with most episodes occurring during the winter months. The virus likely has a worldwide distribution. Almost all children have been infected by five years of age. The suspicion of hMPV infection should be higher in infants or children presenting with symptoms compatible with a viral etiology and in whom screening tests for other common viral pathogens have been negative. Clinical manifestations may be subtle or severe, including life-threatening bronchiolitis or pneumonia. Fever, rhinorrhea, cough, retractions, tachypnea and wheezing are common findings. Bronchiolitis is perhaps the most common manifestation among hospitalized children. Currently, there is no antiviral treatment or vaccine available and management is simply supportive. PMID- 20019853 TI - CPSP Highlights. PMID- 20019854 TI - Community-acquired pneumonia in children. AB - Community acquired pneumonia (CAP) is common in childhood. Viruses account for most cases of CAP during the first two years of life. After this period, bacteria such as Streptococcus pneumoniae, Mycoplasma pneumoniae and Chlamydia pneumoniae become more frequent. CAP symptoms are nonspecific in younger infants, but cough and tachypnea are usually present in older children. Chest x-ray is useful for confirming the diagnosis. Most children can be managed empirically with oral antibiotics as outpatients without specific laboratory investigations. Those with severe infections or with persistent or worsening symptoms need more intensive investigations and may need admission to hospital. The choice and dosage of antibiotics should be based on the age of the patient, severity of the pneumonia and knowledge of local antimicrobial resistance patterns. The Canadian Paediatric Society recommends the use of the heptavalent conjugate pneumococcal vaccine, which is efficacious in reducing chest x-ray positive pneumonia by up to 20%. PMID- 20019855 TI - Influenza vaccination options to prevent hospitalization. AB - BACKGROUND: Vaccination of children against influenza remains a controversial topic despite the substantial morbidity caused by this infection. OBJECTIVE: To estimate the effect of three different vaccination strategies on preventing hospitalization due to influenza. METHODS: A retrospective chart review was conducted of all children admitted to a tertiary health care centre who tested positive for influenza during three consecutive influenza seasons. RESULTS: The final analysis included 208 cases with an age range of five days to 16.1 years. Seventy-six children were considered 'high-risk' and 132 were considered 'previously healthy'. Length of stay (LOS) ranged from one day to 46 days with a mean of 6.3 days. The mean LOS was 8.6 days for children with risk factors and 4.9 days for those without risk factors. The number of preventable influenza admissions was determined over three years and averaged over one year for the three vaccination strategies. A universal strategy of vaccinating all previously healthy and high-risk children over six months of age would have prevented 118 admissions. Using a selective strategy of vaccinating only children over six months of age with risk factors and a third strategy of vaccinating only two- to six-month-old infants would have prevented 58 and 55 admissions, respectively. CLINICAL IMPLICATION: Use of the universal vaccination strategy would have prevented over one-half of the influenza admissions, which was over twice that of targeted vaccination. Until the challenges of implementing universal vaccination are fully understood, targeted vaccination remains an acceptable alternative. PMID- 20019856 TI - Infection control and paediatric tuberculosis: A practical guide for the practicing paediatrician. AB - Tuberculosis (TB) in children requires close attention to infection control to prevent transmission to other patients and health care workers. Although many children with TB are not infectious, appropriate airborne precautions must be maintained until conditions that increase the risk of transmission have been ruled out and accompanying adults, who may also be infectious, have been screened. Concurrent strategies to prevent TB transmission should be implemented, including administrative, engineering and personal protective measures. The most important measure is maintaining a high clinical index of suspicion for TB in patients with compatible symptoms and epidemiological risk factors. Comprehensive tuberculin skin test programmes and the use of N 95 masks can reduce the risk of transmission within health care settings. Current standards of practice should be followed to prevent transmission from patients with active TB disease. PMID- 20019857 TI - In infants with bronchiolitis, is epinephrine more effective than placebo in improving clinical outcomes, as measured by clinical score, oxygen saturation and respiratory rate?: Part A: Evidence-based answer and summary. PMID- 20019858 TI - Use of palivizumab in children with congenital heart disease. PMID- 20019859 TI - Influenza vaccine. PMID- 20019860 TI - Swine flu (H1N1/09): an interim assessment. PMID- 20019861 TI - Early clinical experiences with the new influenza A (H1N1/09). AB - BACKGROUND: Because of ongoing person-to-person transmission of the disease, the World Health Organization has declared a phase 6 pandemic alert for the new type of influenza A (H1N1/09). This means that the spread of the disease must be closely monitored. METHODS: At the Dusseldorf University Hospital, patients with flu-like symptoms and their contacts have been tested for the new type of influenza A since April 2009. RESULTS: The first patients that tested positive for H1N1/09 were treated on 20 May 2009. By mid-September, 3372 persons underwent PCR testing of a sample obtained by deep nasal swabbing, and the results were positive in 450 (13.3%). 379 of these 450 infections, or 84.2%, had been contracted abroad. Most patients came to the hospital with flu-like symptoms within three days of becoming ill. An analysis of the first 60 patients revealed a median core temperature of 37.8 degrees C and a mildly elevated C-reactive protein concentration. All patients were treated with oseltamivir. Most of the initially symptomatic patients were asymptomatic again within 3 days; the median duration of treatment was 5 days. The median time to the first negative deep nasal swab was 4 days. No oseltamivir resistance has been found to date in our patient collective. CONCLUSION: The clinical manifestations of the new type of influenza were still mild in the patient population that we studied up to mid September 2009. At that time, the second wave of the pandemic had not yet begun in Germany. At present, however, the number of cases acquired within the country is on the rise. PMID- 20019862 TI - Influenza--insights from mathematical modelling. AB - BACKGROUND: When the first cases of a new infectious disease appear, questions arise about the further course of the epidemic and about the appropriate interventions to be taken to protect individuals and the public as a whole. Mathematical models can help answer these questions. In this article, the authors describe basic concepts in the mathematical modelling of infectious diseases, illustrate their use with a simple example, and present the results of influenza models. METHOD: Description of the mathematical modelling of infectious diseases and selective review of the literature. RESULTS: The two fundamental concepts of mathematical modelling of infectious diseases-the basic reproduction number and the generation time-allow a better understanding of the course of an epidemic. Modelling studies based on past influenza epidemics suggest that the rise of the epidemic curve can be slowed at the beginning of the epidemic by isolating ill persons and giving prophylactic medications to their contacts. Later on in the course of the epidemic, restricting the number of contacts (e.g., by closing schools) may mitigate the epidemic but will only have a limited effect on the total number of persons who contract the disease. CONCLUSION: Mathematical modelling is a valuable tool for understanding the dynamics of an epidemic and for planning and evaluating interventions. PMID- 20019863 TI - Placebo: misunderstandings and prejudices. AB - BACKGROUND: The role of placebos is often misunderstood, leading both to overvaluation and to inappropriate disdain. The effect of a placebo that contains no pharmacologically active substance is often confused with the effect of administration by a physician. The aim of this article is to review the current data on placebos, evaluate these data critically, and provide a well-founded and understandable explanation of the effects that placebos do and do not possess. METHODS: Selective literature review. RESULTS: Recent studies employing modern imaging techniques have provided objective correlates of the effect of placebo administration for certain indications. A recent paper even suggested a genetic basis for it. Two main mechanisms underlie the effect of placebo administration: conditioned reflexes, which are subconscious, and the patient's expectations, which are conscious. Further factors include the physician's personality and the setting in which the treatment takes place. CONCLUSIONS: The mechanisms of action of placebo administration, with which positive therapeutic effects can be achieved with little effort, should be consciously exploited by physicians when giving their patients pharmacologically active medications as well. PMID- 20019864 TI - Hypocomplementemic urticarial vasculitis syndrome: an interdisciplinary challenge. AB - BACKGROUND: Chronic urticaria often points the way to the diagnosis of a systemic disease, particularly when urticarial vasculitis can be demonstrated. Hypocomplementemic urticarial vasculitis syndrome (HUVS) is considered to be an independent immunological disease. METHOD: Selective literature review and consideration of the author's own clinical experience. RESULTS AND CONCLUSIONS: The main manifestation of HUVS is chronic urticarial vasculitis with complement deficiency and the demonstration of C1q antibody in the serum. Multiple other organs are involved, sometimes severely. The diagnosis is confirmed by skin biopsy, which reveals leukocytoclastic vasculitis as a pathogenetic correlate of this systemic disease. Although HUVS is relatively rare, the medical specialists that might encounter it-ophthalmologists, rheumatologists, nephrologists, dermatologists, general practitioners, and pediatricians-should include it in their differential diagnoses whenever appropriate. Awareness of HUVS and rational diagnostic evaluation will lessen the chance of it being misdiagnosed as another type of systemic immunological disease and will reduce superfluous diagnostic testing in patients suffering from it. PMID- 20019865 TI - Correspondence (letter to the editor): prevention is more cost effective. PMID- 20019867 TI - Correspondence (letter to the editor): obsolescence of X-ray defecography. PMID- 20019868 TI - Correspondence (letter to the editor): a very welcome article. PMID- 20019869 TI - Correspondence (letter to the editor): potential for dependence. PMID- 20019870 TI - Correspondence (letter to the editor): confusion. PMID- 20019871 TI - Correspondence (letter to the editor): an excellent article. PMID- 20019873 TI - An epigenetic signature in peripheral blood predicts active ovarian cancer. AB - BACKGROUND: Recent studies have shown that DNA methylation (DNAm) markers in peripheral blood may hold promise as diagnostic or early detection/risk markers for epithelial cancers. However, to date no study has evaluated the diagnostic and predictive potential of such markers in a large case control cohort and on a genome-wide basis. PRINCIPAL FINDINGS: By performing genome-wide DNAm profiling of a large ovarian cancer case control cohort, we here demonstrate that active ovarian cancer has a significant impact on the DNAm pattern in peripheral blood. Specifically, by measuring the methylation levels of over 27,000 CpGs in blood cells from 148 healthy individuals and 113 age-matched pre-treatment ovarian cancer cases, we derive a DNAm signature that can predict the presence of active ovarian cancer in blind test sets with an AUC of 0.8 (95% CI (0.74-0.87)). We further validate our findings in another independent set of 122 post-treatment cases (AUC = 0.76 (0.72-0.81)). In addition, we provide evidence for a significant number of candidate risk or early detection markers for ovarian cancer. Furthermore, by comparing the pattern of methylation with gene expression data from major blood cell types, we here demonstrate that age and cancer elicit common changes in the composition of peripheral blood, with a myeloid skewing that increases with age and which is further aggravated in the presence of ovarian cancer. Finally, we show that most cancer and age associated methylation variability is found at CpGs located outside of CpG islands. SIGNIFICANCE: Our results underscore the potential of DNAm profiling in peripheral blood as a tool for detection or risk-prediction of epithelial cancers, and warrants further in depth and higher CpG coverage studies to further elucidate this role. PMID- 20019874 TI - Molecular evolution of Drosophila cuticular protein genes. AB - Several multigene families have been described that together encode scores of structural cuticular proteins in Drosophila, although the functional significance of this diversity remains to be explored. Here I investigate the evolutionary histories of several multigene families (CPR, Tweedle, CPLCG, and CPF/CPFL) that vary in age, size, and sequence complexity, using sequenced Drosophila genomes and mosquito outgroups. My objective is to describe the rates and mechanisms of 'cuticle-ome' divergence, in order to identify conserved and rapidly evolving elements. I also investigate potential examples of interlocus gene conversion and concerted evolution within these families during Drosophila evolution. The absolute rate of change in gene number (per million years) is an order of magnitude lower for cuticular protein families within Drosophila than it is among Drosophila and the two mosquito taxa, implying that major transitions in the cuticle proteome have occurred at higher taxonomic levels. Several hotspots of intergenic conversion and/or gene turnover were identified, e.g. some gene pairs have independently undergone intergenic conversion within different lineages. Some gene conversion hotspots were characterized by conversion tracts initiating near nucleotide repeats within coding regions, and similar repeats were found within concertedly evolving cuticular protein genes in Anopheles gambiae. Rates of amino-acid substitution were generally severalfold higher along the branch connecting the Sophophora and Drosophila species groups, and 13 genes have Ka/Ks significantly greater than one along this branch, indicating adaptive divergence. Insect cuticular proteins appear to be a source of adaptive evolution within genera and, at higher taxonomic levels, subject to periods of gene-family expansion and contraction followed by quiescence. However, this relative stasis is belied by hotspots of molecular evolution, particularly concerted evolution, during the diversification of Drosophila. The prominent association between interlocus gene conversion and repeats within the coding sequence of interacting genes suggests that the latter promote strand exchange. PMID- 20019875 TI - One-year follow-up of epithelial corneal cell sheet allografts mounted on platelet poor plasma in rabbits. AB - PURPOSE: To evaluate the usefulness of epithelial corneal sheets mounted on platelet poor plasma (PPP) for allograft transplantation of rabbits with total limbal stem cell deficiency (LSCD) and to prove its efficacy at 1 year after surgery. METHODS: LSCD was induced in 21 female rabbits by mechanical keratectomy. To configure the grafts, limbal biopsies were taken from male rabbits and cells were cultured on a fibroblast feeder layer grown on clotted autologous PPP. After keratectomy, grafts were sutured over the stroma. Control groups consisted of no implant or an implant of clotted PPP. Rabbits were euthanized at 3 and 12 months. Corneas and cultured sheets were processed for histopathology and immunohistochemistry (K3/12 and K19). Gender analysis was performed at 4 and 7 months. RESULTS: One rabbit had endophthalmitis, and another died of no apparent cause. The rest of the animals treated had no inflammation, showed a stratified epithelium, keratin 3/12 expression, and no expression of keratin 19. At 1 year, seven of eight rabbits showed no LSCD or corneal rejection signs. Y chromosomes were detected at 4 and 7 months postoperatively. All controls showed LSCD signs, erratic epithelium, and minimal cell differentiation; they revealed a slight expression of K3/12 and an expression of K19 in patchy patterns. CONCLUSIONS: Allografts contributed to restoring a healthy eye surface without signs of graft rejection. This technique seems to be a promising procedure for bilateral ocular surface diseases and may be useful for new therapeutic strategies. PMID- 20019876 TI - Tryptophan and kynurenine levels in lenses of Wistar and accelerated-senescence OXYS rats. AB - PURPOSE: To determine the levels of kynurenine (KN) and its metabolic precursor tryptophan (Trp) in lenses of accelerated-senescence OXYS (cataract model) and Wistar (control) rats at ages from 1 day to 24 months. METHODS: Protein-free lens extracts were prepared from Wistar and senescent-accelerated OXYS rat lenses. The presence and levels of KN and Trp were determined using high performance liquid chromatography (HPLC) analysis and mass spectrometric measurements. All statistical calculations were made using the software package Statistica 6.0, using factor dispersion analysis and Newman-Keuls post-hoc test for comparison of group mean values. RESULTS: The levels of KN, which plays the role of a molecular Ultraviolet (UV) filter in the human lens, and its metabolic precursor Trp in the rat lens significantly depend on the rat strain and age. During the first 20 days after birth, before the first signs of cataract in OXYS rats, there is a strong difference in the content of both Trp and KN between Wistar and OXYS lenses. This difference becomes insignificant in lenses of 1 month and older. The levels of Trp and KN in young lenses are higher than that in lenses of 1 month and older for both strains. CONCLUSIONS: The presented results demonstrate that the KN pathway of Trp catabolism does not play a significant role in cataract development in the rat lens at the stages of cataract manifestation; however, in the first 3 weeks of postnatal development, the interstrain difference in KN and Trp levels is very strong. The obtained results show a correlation between the low level of KN and the high level of Trp at the stage of lens maturation and future cataractogenesis, and suggest an imbalance in the KN pathway of Trp catabolism in potentially cataractous lenses. PMID- 20019877 TI - Protective effect of clusterin on oxidative stress-induced cell death of human corneal endothelial cells. AB - PURPOSE: To investigate the protective effect of clusterin on oxidative stress induced cell death of human corneal endothelial cells. METHODS: Human corneal endothelial cells (HCECs) were cultured according to previously published methods. With treatment of various concentrations (0-50 mM) of tert-butyl hydroperoxide (tBHP) or clusterin, reactive oxygen species (ROS) producrion was measured using an oxidationsensitive fluorescent probe and 2'7' dichlorofluorescin diacetate (DCFH-DA). Cell viability was assayed with a Cell Counting Kit-8. RESULTS: In HCECs, DCF-DA staining revealed that cells treated with a higher concentration of tBHP had higher fluorescent intensity than cells treated with clusterin, compared to control cells. Clusterin significantly inhibited tBHP-induced ROS production. Cell viability decreased with higher tBHP concentration. Cells treated with clusterin had higher viability than control cells at 5 mM tBHP. Clusterin effectively protected HCECs from ROS-induced cell death. CONCLUSIONS: Our data suggest that clusterin may protect HCECs from oxidative injury-mediated cell death via inhibition of ROS production. PMID- 20019878 TI - Efficient expression of self-complementary AAV in ganglion cells of the ex vivo primate retina. AB - PURPOSE: To evaluate the efficiency of self-complementary adeno-associated virus (scAAV)-mediated gene expression of green fluorescent protein (GFP) or the allotopic human ND4 subunit of complex I in ganglion cells of the primate retina. METHODS: ScAAV2 containing the cDNA encoding the humanized GFP or allotopic ND4 subunit of complex I under the control of the cytomegalovirus (CMV) immediate early gene enhancer and short chicken beta-actin promoter-exon1-intron1 (CBA) was injected into the vitreous cavity of five primate eyes after enucleation. Following incubation in standard Dulbecco's Modified Eagle Medium (DMEM) culture media overnight at 37 degrees C with 5% CO(2), retinal flat mounts were probed with monoclonal GFP or FLAG antibodies overnight followed by counterstaining with anti-mouse IgG conjugated to cy2. For identification of retinal ganglion cells (RGCs), the retinal whole mounts were also stained with a Brn3a or Thy1.2 (protein expressed in RGCs. domain) antibody, then counterstained with cy3 or cy2. Immunofluorescence and colocalization were assessed using confocal microscopy. Quantitative analysis of GFP, ND4FLAG, Brn3a, or Thy1.2 expressing cells was performed using Image J software. RESULTS: While the endogenous fluorescence of GFP was seen in a few retinal cells, GFP and ND4FLAG immunofluorescence was plentiful. The immunosignals were restricted to the inner retina and colocalized to slightly more than half of all cells expressing Brn3a or Thy1.2, suggesting efficient expression in RGCs. CONCLUSIONS: Our findings suggest that the hybrid CMV enhancer-CBetaA promoter can play an efficient role in targeting primate RGCs following intravitreal gene delivery using the scAAV2 vector. Donated ex vivo primate eyes may serve as a model system for testing RGC expression before in vivo intravitreal injections of this and perhaps other AAV serotypes. PMID- 20019879 TI - Immunohistochemical changes in rat retinas at various time periods of elevated intraocular pressure. AB - PURPOSE: To study alterations in different retinal cell types associated with retinal ganglion cell (RGC) death after elevation of intraocular pressure (IOP) in rats. METHODS: IOP was elevated by episcleral vein cauterization of the rat left eye. The right unoperated eye was kept as the control. IOP was measured when rats were awake. The animals were euthanized after one week (n=4) and five weeks (n=4). Their eyes were enucleated, postfixed, cryoprotected, and embedded in optimal cutting temperature (OCT) medium. Cryosections of the retina were cut at 14 microm thickness and processed for immunocytochemistry with 15 antibodies that specifically stain different retinal cell types. The distribution and intensity of the label was analyzed by comparing sections of control and glaucomatous retinas obtained from identical locations. RESULTS: The amount of amacrine cells identified by calcium binding proteins and choline acetyltransferase antibodies decreased after five weeks of elevated IOP. By using the anti-protein kinase C alpha antibody, we were able to label a subpopulation of rod bipolar cells in control retinas but not in retinas that had elevated IOP. No changes were found in RGCs labeled with brain derived neurotrophic factor when comparing control and glaucomatous retinas. Glial fibrillary acidic protein and vimentin expression in glial cells increased after one week of elevated IOP. CONCLUSIONS: After one week of elevated IOP and before the onset of RGC death, it was evident that inner retinal cells showed remarkable changes in their molecular expression. PMID- 20019880 TI - Polymorphisms in the VEGFA and VEGFR-2 genes and neovascular age-related macular degeneration. AB - PURPOSE: Genetic factors influence an individual's risk for developing neovascular age-related macular degeneration (AMD), a leading cause of irreversible blindness. Previous studies on the potential genetic link between AMD and vascular endothelial growth factor (VEGF), a key regulator of angiogenesis and vascular permeability, have yielded conflicting results. In the present case-control association study, we aimed to determine whether VEGF or its main receptor tyrosine kinase VEGFR-2 is genetically associated with neovascular AMD. METHODS: A total of 515 Caucasian patients with neovascular AMD and 253 ethically-matched controls were genotyped for polymorphisms in the VEGFA and VEGFR-2 genes. A tagging single nucleotide polymorphism (tSNP) approach was employed to cover each gene plus two kilobases on each side, spanning the promoter and 3' untranslated regions. SNPs with a minimum allele frequency of 10% were covered by seven tSNPs in VEGFA and 20 tSNPs in VEGFR-2. Two VEGFA SNPs previously linked with AMD, rs1413711 and rs3025039, were also analyzed. RESULTS: The 29 VEGFA and VEGFR-2 SNPs analyzed in our cohort demonstrated no significant association with neovascular AMD. A single rare haplotype in the VEGFR-2 gene was associated with the presence of neovascular AMD (p=0.034). CONCLUSIONS: This study is the first to investigate the association of VEGFR-2 polymorphisms with AMD and evaluates VEGFA genetic variants in the largest neovascular AMD cohort to date. Despite the angiogenic and permeability-enhancing effects of VEGF/VEGFR-2 signaling, we found minimal evidence of a significant link between polymorphisms in the VEGFA and VEGFR-2 genes and neovascular AMD. PMID- 20019881 TI - Microarray analysis of retinal gene expression in Egr-1 knockout mice. AB - PURPOSE: We found earlier that 42 day-old Egr-1 knockout mice had longer eyes and a more myopic refractive error compared to their wild-types. To identify genes that could be responsible for the temporarily enhanced axial eye growth, a microarray analysis was performed in knockout and wild-type mice at the postnatal ages of 30 and 42 days. METHODS: The retinas of homozygous and wild-type Egr-1 knockout mice (Taconic, Ry, Denmark) were prepared for RNA isolation (RNeasy Mini Kit, Qiagen) at the age of 30 or 42 days, respectively (n=12 each). Three retinas were pooled and labeled cRNA was made. The samples were hybridized to Affymetrix GeneChip Mouse Genome 430 2.0 Arrays. Hybridization signals were calculated using GC-RMA normalization. Genes were identified as differentially expressed if they showed a fold-change (FC) of at least 1.5 and a p-value <0.05. A false-discovery rate of 5% was applied. Ten genes with potential biologic relevance were examined further with semiquantitative real-time RT-PCR. RESULTS: Comparing mRNA expression levels between wild-type and homozygous Egr-1 knockout mice, we found 73 differentially expressed genes at the age of 30 days and 135 genes at the age of 42 days. Testing for differences in gene expression between the two ages (30 versus 42 days), 54 genes were differently expressed in wild-type mice and 215 genes in homozygous animals. Based on three networks proposed by Ingenuity pathway analysis software, nine differently expressed genes in the homozygous Egr 1 knockout mice were chosen for further validation by real-time RT-PCR, three genes in each network. In addition, the gene that was most prominently regulated in the knockout mice, compared to wild-type, at both 30 days and 42 days of age (protocadherin beta-9 [Pcdhb9]), was tested with real-time RT-PCR. Changes in four of the ten genes could be confirmed by real-time RT-PCR: nuclear prelamin A recognition factor (Narf), oxoglutarate dehydrogenase (Ogdh), selenium binding protein 1 (Selenbp1), and Pcdhb9. Except for Pcdhb9, the genes whose mRNA expression levels were validated were listed in one of the networks proposed by Ingenuity pathway analysis software. In addition to these genes, the software proposed several key-regulators which did not change in our study: retinoic acid, vascular endothelial growth factor A (VEGF-A), FBJ murine osteosarcoma viral oncogene homolog (cFos), and others. CONCLUSIONS: Identification of genes that are differentially regulated during the development period between postnatal day 30 (when both homozygous and wild-type mice still have the same axial length) and day 42 (where the difference in eye length is apparent) could improve the understanding of mechanisms for the control of axial eye growth and may lead to potential targets for pharmacological intervention. With the aid of pathway analysis software, a coarse picture of possible biochemical pathways could be generated. Although the mRNA expression levels of proteins proposed by the software, like VEGF, FOS, retinoic acid (RA) receptors, or cellular RA binding protein, did not show any changes in our experiment, these molecules have previously been implicated in the signaling cascades controlling axial eye growth. According to the pathway-analysis software, they represent links between several proteins whose mRNA expression was changed in our study. PMID- 20019882 TI - Functional proteomics of failed filtering blebs. AB - PURPOSE: To identify and determine the function of the proteins associated with failed filtering blebs following trabeculectomy. METHODS: Tenon's tissues, obtained during surgery for failed filtering blebs or obtained during cataract surgery on normal eyes, were analyzed by proteomics. The proteins showing significant differences between the two tissues were selected for identification by mass spectrometry. The location and expression pattern of ribosomal S6 kinase 2 (RSK2), one of the altered proteins, were determined. The effect of basic fibroblast growth factor (bFGF) on the expression pattern and function of RSK2 in NIH3T3 fibroblast cells was then investigated by an RNA knockdown technique. RESULTS: Eight proteins were found differentially expressed in failed filtering blebs; the identified proteins included those associated with intracellular signaling pathways. The expression of RSK2, one of the identified proteins, was found to be decreased compared with that of the control. RSK2 was located in Tenon's tissue using an immunohistochemical technique. In culture, the bFGF induced cell proliferation was inhibited by the RNA knockdown of RSK2. The level of mRNA and protein expression of actin was increased by RSK2 RNA knockdown, but bFGF-induced protein expression of actin was not promoted by RSK2 RNA knockdown. Whereas RSK2 RNA knockdown increased the expression and activity of mitogen activated protein kinase (MAPK), activation of MAPK induced by bFGF was not promoted by RSK2 knockdown. CONCLUSIONS: The expression of eight proteins in the failed filtering blebs was significantly different from that in the Tenon's capsules used as a control. The effect of RSK2 expression on fibroblast cells suggests that RSK2 may be associated with wound healing in filtering blebs. PMID- 20019883 TI - Retinal phototoxicity in a novel murine model of intraocular lens implantation. AB - PURPOSE: To establish a novel murine intraocular lens (IOL) implantation model to study the protective effects of colored-IOLs against retinal phototoxicity. METHODS: Two-millimeter diameter IOL buttons were created from IOLs for clinical use. Extra-capsular crystalline lens extraction and IOL implantation were performed in BALB/c mice using a technique similar to human cataract surgery. For light exposure experiments, mice were exposed to 5,000 LUX of white light for 24 h on the day after surgery. To investigate the protective effects of yellow IOL against light exposure, ERG measurements were conducted in vivo, followed by TdT mediated dUTP Nick-End Labeling (TUNEL) and outer nuclear layer (ONL) thickness measurement of retinal tissue in yellow or clear IOL-implanted mice and control mice without surgery. RESULTS: IOLs were successfully implanted in all animals, and IOL buttons without haptics were well stabilized in the capsular bag. Murine eyes developed posterior capsule opacification (PCO) after IOL implantation by postoperative day 5 at the latest. In contrast to the clear IOL-implanted animals stimulated by light exposure, the yellow IOL-implanted animals had significantly reduced numbers of TUNEL-positive cells and retained thickness of the ONL. The ERG showed that yellow IOL implantation prevents a decrease of amplitude in both the a-wave and b-wave compared with clear IOL implantation. CONCLUSIONS: We established a new animal model of IOL implantation and demonstrated the protective effects of colored-IOL against retinal phototoxicity after cataract surgery. PMID- 20019884 TI - Proteomic analysis of human aqueous humor using multidimensional protein identification technology. AB - Aqueous humor (AH) supports avascular tissues in the anterior segment of the eye, maintains intraocular pressure, and potentially influences the pathogenesis of ocular diseases. Nevertheless, the AH proteome is still poorly defined despite several previous efforts, which were hindered by interfering high abundance proteins, inadequate animal models, and limited proteomic technologies. To facilitate future investigations into AH function, the AH proteome was extensively characterized using an advanced proteomic approach. Samples from patients undergoing cataract surgery were pooled and depleted of interfering abundant proteins and thereby divided into two fractions: albumin-bound and albumin-depleted. Multidimensional Protein Identification Technology (MudPIT) was utilized for each fraction; this incorporates strong cation exchange chromatography to reduce sample complexity before reversed-phase liquid chromatography and tandem mass spectrometric analysis. Twelve proteins had multi peptide, high confidence identifications in the albumin-bound fraction and 50 proteins had multi-peptide, high confidence identifications in the albumin depleted fraction. Gene ontological analyses were performed to determine which cellular components and functions were enriched. Many proteins were previously identified in the AH and for several their potential role in the AH has been investigated; however, the majority of identified proteins were novel and only speculative roles can be suggested. The AH was abundant in anti-oxidant and immunoregulatory proteins as well as anti-angiogenic proteins, which may be involved in maintaining the avascular tissues. This is the first known report to extensively characterize and describe the human AH proteome and lays the foundation for future work regarding its function in homeostatic and pathologic states. PMID- 20019885 TI - Cardiovascular Diseases, Aging and the Gender Gap in the Human Longevity. AB - This brief communication probes into the biological meaning of the gender gap in longevity and its possible ramifications to the expression of cardiovascular diseases in humans. It addresses the potential role of the estrogen and the X chromosome in the longer life span of women than men in modern societies. In addition, it links features of the reproductive and post-reproductive periods with cardiovascular diseases and longevity in women. PMID- 20019886 TI - Molecular MRI of Atherosclerotic Plaque With Targeted Contrast Agents. AB - Molecular MRI of atherosclerosis involves the use of novel contrast agents to image cellular and molecular processes within atherosclerotic plaque. Agents to image plaque lipid content, inflammation, angiogenesis, and thrombosis have been developed and studied extensively in animal models of atherosclerosis and vascular injury. Selected agents have also been studied in humans, with highly promising initial results. In this brief review, recent advances as well as opportunities and challenges in the field are discussed. PMID- 20019887 TI - Marginal Mean Models for Dynamic Regimes. AB - A dynamic treatment regime is a list of rules for how the level of treatment will be tailored through time to an individual's changing severity. In general, individuals who receive the highest level of treatment are the individuals with the greatest severity and need for treatment. Thus there is planned selection of the treatment dose. In addition to the planned selection mandated by the treatment rules, the use of staff judgment results in unplanned selection of the treatment level. Given observational longitudinal data or data in which there is unplanned selection, of the treatment level, the methodology proposed here allows the estimation of a mean response to a dynamic treatment regime under the assumption of sequential randomization. PMID- 20019888 TI - The Noc-domain containing C-terminus of Noc4p mediates both formation of the Noc4p-Nop14p submodule and its incorporation into the SSU processome. AB - Noc1p, Noc3p and Noc4p are eukaryotic proteins which play essential roles in yeast ribosome biogenesis and contain a homologous stretch of about 45 aminoacids (Noc-domain) of unknown function. Yeast Noc4p is a component of the small ribosomal subunit (SSU) processome, can be isolated as a stable Noc4p-Nop14p SSU processome submodule from yeast cells, and is required for nuclear steps of small ribosomal subunit rRNA maturation. We expressed a series of mutated alleles of NOC4 in yeast cells and analysed whether the corresponding protein variants support vegetative growth, interact with Nop14p, and are incorporated into the SSU-processome. The data reveal that the essential C-terminus of Noc4p which contains 237 aminoacids including the Noc-domain represents a protein-protein interaction module. It is required and sufficient for its association with Nop14p and several nuclear precursors of the small ribosomal subunit. The N-terminal Noc4-part seems to be targeted to pre-ribosomes via the C-terminus of Noc4p and plays there an essential role in SSU-processome function. Replacement of the Noc4p-Noc-domain by its homologues Noc1p-counterpart results in a hybrid Noc4p variant which fails to associate with Nop14p and pre-ribosomes. On the other hand, exchange of 6 amino acids in the Noc1-Noc-domain of this hybrid Noc4p protein is sufficient to restore its essential in vivo functions. These data suggest that Noc-domains of Noc1p and Noc4p share a common structural backbone in which diverging amino acids play crucial roles in mediating specific regulated interactions. Our analysis allows us to distinguish between different functions of certain domains within Noc4p and contribute to the understanding of how incorporation of Noc4p into ribosomal precursors is coupled to rRNA processing and maturation of the small ribosomal subunit. PMID- 20019889 TI - Relations between Mattering to Step- and Non-Residential Fathers and Adolescent Mental Health. AB - This study examined the relations between perceptions of 133 early adolescents in stepfamilies concerning how much they mattered to their stepfathers and nonresidential biological fathers and adolescents' mental health problems. Mattering to nonresidential biological fathers significantly negatively predicted mother-, teacher-, and youth-reported internalizing problems. Mattering to stepfathers significantly negatively predicted youth-reported internalizing and stepfather- and youth- reported externalizing problems. For teacher-reported externalizing problems, mattering to stepfathers and nonresidential biological fathers significantly interacted. Mattering to either father predicted low externalizing problems; perceptions of mattering to the second father did not predict a further reduction in problems. Results suggest that mattering is an important aspect of father-adolescent relationships, and highlight the importance of considering adolescents' relationships with both nonresidential fathers and stepfathers. PMID- 20019890 TI - Tracking Activities in Complex Settings Using Smart Environment Technologies. AB - The pervasive sensing technologies found in smart homes offer unprecedented opportunities for providing health monitoring and assistance to individuals experiencing difficulties living independently at home. A primary challenge that needs to be tackled to meet this need is the ability to recognize and track functional activities that people perform in their own homes and everyday settings. In this paper we look at approaches to perform real-time recognition of Activities of Daily Living. We enhance other related research efforts to develop approaches that are effective when activities are interrupted and interleaved. To evaluate the accuracy of our recognition algorithms we assess them using real data collected from participants performing activities in our on-campus smart apartment testbed. PMID- 20019892 TI - FcRn receptor-mediated pharmacokinetics of therapeutic IgG in the eye. AB - PURPOSE: The goal of this study was to determine the role of the neonatal Fc (FcRn) receptor in eliminating intravitreally administered full-length immunoglobulin G (IgG) across the blood-retinal barrier. METHODS: FcRn receptor expression in normal and laser photocoagulated retinas was compared quantitatively by real-time RT-PCR. The distribution of intravitreally administered full-length IgG was investigated and compared in wild-type and FcRn knockout mouse eyes as well as normal and laser-photocoagulated rat eyes at several time points. Additionally, the pharmacokinetics of intravitreally injected full-length IgG and chicken immunoglobulin Y (IgY) was compared in the normal rat retina. RESULTS: Intravitreally administered full-length IgG overcame the inner limiting membrane and diffused into the deeper retinal structures in both normal and laser-photocoagulated retinas. Interestingly, IgG was eliminated across the blood-retinal barrier into the blood system in the normal retina, whereas IgY was not. In addition, full-length IgGs did not penetrate across the blood-retinal barrier in the FcRn knockout mouse. Intravitreally injected IgGs were eliminated into the blood system more rapidly in laser-photocoagulated eyes when compared to normal control eyes because of FcRn receptor upregulation in the laser-photocoagulated retina. CONCLUSIONS: FcRn plays an important role in eliminating intravitreally administered full-length IgGs across the blood-retina barrier into the systemic blood system. PMID- 20019893 TI - A novel GJA8 mutation (p.I31T) causing autosomal dominant congenital cataract in a Chinese family. AB - PURPOSE: To identify the genetic defect associated with autosomal dominant congenital nuclear cataract in a Chinese family. METHODS: Family history and clinical data were recorded. The genomic DNA was extracted from peripheral blood leukocytes. All the members were genotyped with microsatellite markers at loci considered to be associated with cataracts. Two-point logarithm of odds (LOD) scores were calculated by using the Linkage software after genotyping. Mutations were detected by DNA sequence analysis of the candidate genes. Effects of amino acid changes on the structure and function of proteins were predicted by bioinformatics analysis. RESULTS: Evidence of a linkage was obtained at markers D1S514 (LOD score [Z]=3.48, recombination fraction [theta]=0.0) and D1S1595 (Z=2.49, theta=0.0). Haplotype analysis indicated that the cataract gene was close to these two markers. Sequencing of the connexin 50 (GJA8) gene revealed a T>C transition at nucleotide position c.92. This nucleotide change resulted in the substitution of highly conserved isoleucine by threonine at codon 31(I31T). This mutation co-segregated with all affected individuals and was not observed in unaffected or 110 normal unrelated individuals. Bioinformatics analysis showed that a highly conserved region was located at Ile31, and the mutation was predicted to affect the function and secondary structure of the GJA8 protein. CONCLUSION: A novel mutation in GJA8 was detected in a Chinese family with autosomal dominant congenital nuclear cataract, providing clear evidence of a relationship between the genotype and the corresponding cataract phenotype. PMID- 20019894 TI - Malaria control insecticide residues in breast milk: the need to consider infant health risks. AB - BACKGROUND: In many parts of the world, deliberate indoor residual spraying (IRS) of dwellings with insecticides to control malaria transmission remains the only viable option, thereby unintentionally but inevitably also causing exposure to inhabitants. Because mothers are exposed to insecticides via various routes, accumulated residues are transferred to infants via breast milk, in some cases exceeding recommended intake levels. Except for dichlorodiphenyltrichloroethane (DDT), safety of residues of other insecticides in breast milk has not been considered during World Health Organization Pesticide Evaluation Scheme (WHOPES) evaluations. However, very little is known of the health risks posed by these chemicals to infants who, in developing countries, breast-feed for up to 2 years. OBJECTIVE: We evaluated the need for WHOPES to include breast milk as a potentially significant route of exposure and risk to infants when evaluating the risks during evaluation of IRS insecticides. DISCUSSION: We present evidence showing that neurologic and endocrine effects are associated with pyrethroids and DDT at levels equal or below known levels in breast milk. CONCLUSIONS: Because millions of people in malaria control areas experience conditions of multiple sources and routes of exposure to any number of insecticides, even though lives are saved through malaria prevention, identification of potential infant health risks associated with insecticide residues in breast milk must be incorporated in WHOPES evaluations and in the development of appropriate risk assessment tools. PMID- 20019895 TI - What additional factors beyond state-of-the-art analytical methods are needed for optimal generation and interpretation of biomonitoring data? AB - BACKGROUND: The routine use of biomonitoring (i.e., measurement of environmental chemicals, their metabolites, or specific reaction products in human biological specimens) to assess internal exposure (i.e., body burden) has gained importance in exposure assessment. OBJECTIVES: Selection and validation of biomarkers of exposure are critical factors in interpreting biomonitoring data. Moreover, the strong relation between quality of the analytical methods used for biomonitoring and quality of the resulting data is well understood. However, the relevance of collecting, storing, processing, and transporting the samples to the laboratory to the overall biomonitoring process has received limited attention, especially for organic chemicals. DISCUSSION: We present examples to illustrate potential sources of unintended contamination of the biological specimen during collection or processing procedures. The examples also highlight the importance of ensuring that the biological specimen analyzed both represents the sample collected for biomonitoring purposes and reflects the exposure of interest. CONCLUSIONS: Besides using high-quality analytical methods and good laboratory practices for biomonitoring, evaluation of the collection and handling of biological samples should be emphasized, because these procedures can affect the samples integrity and representativeness. Biomonitoring programs would be strengthened with the inclusion of field blanks. PMID- 20019896 TI - Chlorination disinfection by-products in drinking water and congenital anomalies: review and meta-analyses. AB - OBJECTIVES: The aim of this study was to review epidemiologic evidence, provide summary risk estimates of the association between exposure to chlorination disinfection by-products (DBPs) and congenital anomalies, and provide recommendations for future studies. DATA SOURCES AND EXTRACTION: We included all published epidemiologic studies that evaluated a relationship between an index of DBP exposure (treatment, water source, DBP measurements, and both DBP measurements and personal characteristics) and risk of congenital anomalies. When three or more studies examined the same exposure index and congenital anomaly, we conducted a meta-analysis to obtain a summary risk estimate comparing the highest exposure group with the lowest exposure group. When five or more studies examined total trihalomethane (TTHM) exposure and a specific congenital anomaly, we conducted a meta-analysis to obtain exposure-response risk estimates per 10 microg/L TTHM. DATA SYNTHESIS: For all congenital anomalies combined, the meta analysis gave a statistically significant excess risk for high versus low exposure to water chlorination or TTHM [17%; 95% confidence interval (CI), 3-34] based on a small number of studies. The meta-analysis also suggested a statistically significant excess risk for ventricular septal defects (58%; 95% CI, 21-107), but this was based on only three studies, and there was little evidence of an exposure-response relationship. We observed no statistically significant relationships in the other meta-analyses. We found little evidence for publication bias, except for urinary tract defects and cleft lip and palate. CONCLUSION: Although some individual studies have suggested an association between chlorination disinfection by-products and congenital anomalies, meta analyses of all currently available studies demonstrate little evidence of such an association. PMID- 20019897 TI - Using national and local extant data to characterize environmental exposures in the national children's study: Queens County, New York. AB - OBJECTIVE: The National Children's Study is a long-term epidemiologic study of 100,000 children from 105 locations across the United States. It will require information on a large number of environmental variables to address its core hypotheses. The resources available to collect actual home and personal exposure samples are limited, with most of the home sampling completed on periodic visits and the personal sampling generally limited to biomonitoring. To fill major data gaps, extant data will be required for each study location. The Queens Vanguard Center has examined the extent of those needs and the types of data that are generally and possibly locally available. DATA: In this review we identify three levels of data--national, state and county--and local data and information sets (levels 1-3, respectively), each with different degrees of availability and completeness, that can be used as a starting point for the extant data collection in each study location over time. We present an example on the use of this tiered approach, to tailor the data needs for Queens County and to provide general guidance for application to other NCS locations. CONCLUSIONS: Preexisting and continually evolving databases are available for use in the NCS to characterize exposure. The three levels of data we identified will be used to test a method for developing exposure indices for segments and homes during the pilot phase of NCS, as outlined in this article. PMID- 20019898 TI - A systematic review and meta-analysis of childhood leukemia and parental occupational pesticide exposure. AB - OBJECTIVES: We conducted a systematic review and meta-analysis of childhood leukemia and parental occupational pesticide exposure. DATA SOURCES: Searches of MEDLINE (1950-2009) and other electronic databases yielded 31 included studies. DATA EXTRACTION: Two authors independently abstracted data and assessed the quality of each study. DATA SYNTHESIS: Random effects models were used to obtain summary odds ratios (ORs) and 95% confidence intervals (CIs). There was no overall association between childhood leukemia and any paternal occupational pesticide exposure (OR = 1.09; 95% CI, 0.88-1.34); there were slightly elevated risks in subgroups of studies with low total-quality scores (OR = 1.39; 95% CI, 0.99-1.95), ill-defined exposure time windows (OR = 1.36; 95% CI, 1.00-1.85), and exposure information collected after offspring leukemia diagnosis (OR = 1.34; 95% CI, 1.05-1.70). Childhood leukemia was associated with prenatal maternal occupational pesticide exposure (OR = 2.09; 95% CI, 1.51-2.88); this association was slightly stronger for studies with high exposure-measurement-quality scores (OR = 2.45; 95% CI, 1.68-3.58), higher confounder control scores (OR = 2.38; 95% CI, 1.56-3.62), and farm-related exposures (OR = 2.44; 95% CI, 1.53-3.89). Childhood leukemia risk was also elevated for prenatal maternal occupational exposure to insecticides (OR = 2.72; 95% CI, 1.47-5.04) and herbicides (OR = 3.62; 95% CI, 1.28-10.3). CONCLUSIONS: Childhood leukemia was associated with prenatal maternal occupational pesticide exposure in analyses of all studies combined and in several subgroups. Associations with paternal occupational pesticide exposure were weaker and less consistent. Research needs include improved pesticide exposure indices, continued follow-up of existing cohorts, genetic susceptibility assessment, and basic research on childhood leukemia initiation and progression. PMID- 20019899 TI - Prediagnostic serum concentrations of organochlorine compounds and risk of testicular germ cell tumors. AB - BACKGROUND: Recent findings suggest that exposure to organochlorine (OC) compounds, chlordanes and p,p'-dichlorodiphenyldichloroethylene (p,p'-DDE) in particular, may increase the risk of developing testicular germ cell tumors (TGCTs). OBJECTIVE: To further investigate this question, we conducted a nested case-control study of TGCTs within the Norwegian Janus Serum Bank cohort. METHODS: The study was conducted among individuals with serum collected between 1972 and 1978. TGCT cases diagnosed through 1999 (n = 49; 27-62 years of age at diagnosis) were identified through linkage to the Norwegian Cancer Registry. Controls (n =51) were matched to cases on region, blood draw year, and age at blood draw. Measurements of 11 OC insecticide compounds and 34 polychlorinated biphenyl (PCB) congeners were performed using gas chromatography/high-resolution mass spectrometry. Case-control comparisons of lipid-adjusted analyte concentrations were performed using the Wilcoxon signed-rank test. Odds ratios (ORs) and 95% confidence intervals (CIs) for tertiles of analyte concentration were calculated using conditional logistic regression. RESULTS: TGCT cases had elevated concentrations of p,p'-DDE (tertile 3 vs. tertile 1 OR (OR(T3)) 2.2; 95% CI, 0.7-6.5; p(Wilcoxon) = 0.07), oxychlordane (OR(T3) 3.2; 95% CI, 0.6-16.8; p(Wilcoxon) = 0.05), trans-nonachlor (OR(T3) 2.6; 95% CI, 0.7-8.9; p(Wilcoxon) = 0.07), and total chlordanes (OR(T3) 2.0; 95% CI, 0.6-7.2; p(Wilcoxon) = 0.048) compared with controls, although no ORs were statistically significant. Seminoma cases had significantly lower concentrations of PCB congeners 44, 49, and 52 and significantly higher concentrations of PCBs 99, 138, 153, 167, 183, and 195. CONCLUSIONS: Our study provides additional but qualified evidence supporting an association between exposures to p,p'-DDE and chlordane compounds, and possibly some PCB congeners, and TGCT risk. PMID- 20019900 TI - Diet contributes significantly to the body burden of PBDEs in the general U.S. population. AB - BACKGROUND: Exposure of the U.S. population to polybrominated diphenyl ethers (PBDEs) is thought to be via exposure to dust and diet. However, little work has been done to empirically link body burdens of these compounds to either route of exposure. OBJECTIVES: The primary goal of this research was to evaluate the dietary contribution to PBDE body burdens in the United States by linking serum levels to food intake. METHODS: We used two dietary instruments--a 24-hr food recall (24FR) and a 1-year food frequency questionnaire (FFQ)--to examine food intake among participants of the 2003-2004 National Health and Nutrition Examination Survey. We regressed serum concentrations of five PBDEs (BDE congeners 28, 47, 99, 100, and 153) and their sum (Sigma PBDE) against diet variables while adjusting for age, sex, race/ethnicity, income, and body mass index. RESULTS: Sigma PBDE serum concentrations among vegetarians were 23% (p = 0.006) and 27% (p = 0.009) lower than among omnivores for 24FR and 1-year FFQ, respectively. Serum levels of five PBDE congeners were associated with consumption of poultry fat: Low, medium, and high intake corresponded to geometric mean Sigma PBDE concentrations of 40.6, 41.9, and 48.3 ng/g lipid, respectively (p = 0.0005). We observed similar trends for red meat fat, which were statistically significant for BDE-100 and BDE-153. No association was observed between serum PBDEs and consumption of dairy or fish. Results were similar for both dietary instruments but were more robust using 24FR. CONCLUSIONS: Intake of contaminated poultry and red meat contributes significantly to PBDE body burdens in the United States. PMID- 20019901 TI - Maternal blood lead levels and the risk of pregnancy-induced hypertension: the EDEN cohort study. AB - BACKGROUND: Prior studies revealed associations of environmental lead exposure with risks of hypertension and elevated blood pressure. OBJECTIVE: We examined the effect of blood lead levels on blood pressure and the incidence of pregnancy induced hypertension (PIH) in the second and third trimesters of pregnancy. METHODS: One thousand seventeen pregnant women were enrolled in two French municipalities between 2003 and 2005 for the EDEN (Etude des Determinants pre et post natals du developpement et de la sante de l' Enfant) cohort study. Blood lead concentrations were measured by atomic absorption spectrometry in mothers between 24 and 28 weeks of gestation. RESULTS: PIH was diagnosed in 106 subjects (10.9%). Age, parity, weight gain, alcohol, smoking habits, and calcium supplementation were comparable between hypertensive and nonhypertensive women. Lead levels were significantly higher in PIH cases (mean +/- SD, 2.2 +/- 1.4 microg/dL) than in normotensive patients (1.9 +/- 1.2 microg/dL; p = 0.02). Adjustment for potential confounder effects slightly attenuated but did not eliminate the significant association between blood lead levels and the risk of PIH (adjusted odds ratio of PIH = 3.3; 95% confidence interval, 1.1-9.7). We also observed geographic differences in lead exposure and in the incidence of PIH and found significant correlations between blood lead levels and unadjusted as well as adjusted systolic and diastolic blood pressures after 24 weeks of gestation. CONCLUSIONS: These findings confirm the relationship between blood lead levels at mid-pregnancy and blood pressure and suggest that environmental lead exposure may play an etiologic role in PIH. PMID- 20019903 TI - Mass lead intoxication from informal used lead-acid battery recycling in dakar, senegal. AB - BACKGROUND AND OBJECTIVES: Between November 2007 and March 2008, 18 children died from a rapidly progressive central nervous system disease of unexplained origin in a community involved in the recycling of used lead-acid batteries (ULAB) in the suburbs of Dakar, Senegal. We investigated the cause of these deaths. METHODS: Because autopsies were not possible, the investigation centered on clinical and laboratory assessments performed on 32 siblings of deceased children and 23 mothers and on 18 children and 8 adults living in the same area, complemented by environmental health investigations. RESULTS: All 81 individuals investigated were poisoned with lead, some of them severely. The blood lead level of the 50 children tested ranged from 39.8 to 613.9 microg/dL with a mean of 129.5 microg/dL. Seventeen children showed severe neurologic features of toxicity. Homes and soil in surrounding areas were heavily contaminated with lead (indoors, up to 14,000 mg/kg; outdoors, up to 302,000 mg/kg) as a result of informal ULAB recycling. CONCLUSIONS: Our investigations revealed a mass lead intoxication that occurred through inhalation and ingestion of soil and dust heavily contaminated with lead as a result of informal and unsafe ULAB recycling. Circumstantial evidence suggested that most or all of the 18 deaths were due to encephalopathy resulting from severe lead intoxication. Findings also suggest that most habitants of the contaminated area, estimated at 950, are also likely to be poisoned. This highlights the severe health risks posed by informal ULAB recycling, in particular in developing countries, and emphasizes the need to strengthen national and international efforts to address this global public health problem. PMID- 20019902 TI - Cumulative lead exposure and tooth loss in men: the normative aging study. AB - BACKGROUND: Individuals previously exposed to lead remain at risk because of endogenous release of lead stored in their skeletal compartments. However, it is not known if long-term cumulative lead exposure is a risk factor for tooth loss. OBJECTIVES: We examined the association of bone lead concentrations with loss of natural teeth. METHODS: We examined 333 men enrolled in the Veterans Affairs Normative Aging Study. We used a validated K-shell X-ray fluorescence (KXRF) method to measure lead concentrations in the tibial midshaft and patella. A dentist recorded the number of teeth remaining, and tooth loss was categorized as 0, 1-8 or > or = 9 missing teeth. We used proportional odds models to estimate the association of bone lead biomarkers with tooth loss, adjusting for age, smoking, diabetes, and other putative confounders. RESULTS: Participants with > or = 9 missing teeth had significantly higher bone lead concentrations than those who had not experienced tooth loss. In multivariable-adjusted analyses, men in the highest tertile of tibia lead (> 23 microg/g) and patella lead (> 36 microg/g) had approximately three times the odds of having experienced an elevated degree of tooth loss (> or = 9 vs. 0-8 missing teeth or > or = 1 vs. 0 missing teeth) as those in the lowest tertile [prevalence odds ratio (OR) = 3.03; 95% confidence interval (CI), 1.60-5.76 and OR = 2.41; 95% CI, 1.30-4.49, respectively]. Associations between bone lead biomarkers and tooth loss were similar in magnitude to the increased odds observed in participants who were current smokers. CONCLUSION: Long-term cumulative lead exposure is associated with increased odds of tooth loss. PMID- 20019904 TI - Molecular population genetics of human CYP3A locus: signatures of positive selection and implications for evolutionary environmental medicine. AB - BACKGROUND: The human CYP3A gene cluster codes for cytochrome P450 (CYP) subfamily enzymes that catalyze the metabolism of various exogenous and endogenous chemicals and is an obvious candidate for evolutionary and environmental genomic study. Functional variants in the CYP3A locus may have undergone a selective sweep in response to various environmental conditions. OBJECTIVE: The goal of this study was to profile the allelic structure across the human CYP3A locus and investigate natural selection on that locus. METHODS: From the CYP3A locus spanning 231 kb, we resequenced 54 genomic DNA fragments (a total of 43,675 bases) spanning four genes (CYP3A4, CYP3A5, CYP3A7, and CYP3A43) and two pseudogenes (CYP3AP1 and CYP3AP2), and randomly selected intergenic regions at the CYP3A locus in Africans (24 individuals), Caucasians (24 individuals), and Chinese (29 individuals). We comprehensively investigated the nucleotide diversity and haplotype structure and examined the possible role of natural selection in shaping the sequence variation throughout the gene cluster. RESULTS: Neutrality tests with Tajima's D, Fu and Li's D* and F*, and Fay and Wu's H indicated possible roles of positive selection on the entire CYP3A locus in non Africans. Sliding-window analyses of nucleotide diversity and frequency spectrum, as well as haplotype diversity and phylogenetically inferred haplotype structure, revealed that CYP3A4 and CYP3A7 had recently undergone or were undergoing a selective sweep in all three populations, whereas CYP3A43 and CYP3A5 were undergoing a selective sweep in non-Africans and Caucasians, respectively. CONCLUSION: The refined allelic architecture and selection spectrum for the human CYP3A locus highlight that evolutionary dynamics of molecular adaptation may underlie the phenotypic variation of the xenobiotic disposition system and varied predisposition to complex disorders in which xenobiotics play a role. PMID- 20019905 TI - Perinatal exposure to bisphenol a alters early adipogenesis in the rat. AB - BACKGROUND: The causes of the current obesity pandemic have not been fully elucidated. Implication of environmental endocrine disruptors such as bisphenol A (BPA) on adipose tissue development has been poorly investigated. OBJECTIVES: The aim of the present study was to evaluate the effects of perinatal exposure to BPA on early adipose storage at weaning. METHODS: Pregnant Sprague-Dawley rats had access to drinking water containing 1 mg/L BPA from day 6 of gestation through the end of lactation. Pups were weaned on postnatal day (PND) 21. At that time, we investigated perigonadal adipose tissue of pups (weight, histology, gene expression). For the remaining animals, we recorded body weight and food intake for animals on either standard chow or a high-fat diet. RESULTS: Gestational exposure to BPA did not alter the sex ratio or litter size at birth. On PND1, the weight of male and female BPA-exposed pups was increased. On PND21, body weight was increased only in females, in which parametrial white adipose tissue (pWAT) weight was increased about 3-fold. This excess of pWAT was associated with adipocyte hypertrophy and overexpression of lipogenic genes such as C/EBP-alpha (CAAT enhancer binding protein alpha), PPAR-gamma (peroxisome proliferator activated receptor gamma), SREBP-1C (sterol regulatory element binding protein 1C), LPL (lipoprotein lipase), FAS (fatty acid synthase), and SCD-1 (stearoyl-CoA desaturase 1). In addition, gene expression of SREBP-1C, FAS, and ACC (acetyl-CoA carboxylase) was also increased in liver from BPA-exposed females at PND21, without a change in circulating lipids and glucose. After weaning, perinatal BPA exposure predisposed to overweight in a sex- and diet-dependent manner. We observed no change in food intake due to perinatal BPA exposure in rats on either standard chow or a high-fat diet. CONCLUSIONS: Perinatal exposure to a low dose of BPA increased adipogenesis in females at weaning. Adult body weight may be programmed during early life, leading to changes dependent on the sex and the nutritional status. Although further studies are required to understand the mechanisms of BPA action in early life, these results are particularly important with regard to the increasing prevalence of childhood obesity and the context dependent action of endocrine disruptors. PMID- 20019906 TI - Exposure to a complex cocktail of environmental endocrine-disrupting compounds disturbs the kisspeptin/GPR54 system in ovine hypothalamus and pituitary gland. AB - BACKGROUND: Ubiquitous environmental chemicals, including endocrine-disrupting chemicals (EDCs), are associated with declining human reproductive health, as well as an increasing incidence of cancers of the reproductive system. Verifying such links requires animal models exposed to "real-life," environmentally relevant concentrations/mixtures of EDC, particularly in utero, when sensitivity to EDC exposure is maximal. OBJECTIVES: We evaluated the effects of maternal exposure to a pollutant cocktail (sewage sludge) on the ovine fetal reproductive neuroendocrine axes, particularly the kisspeptin (KiSS-1)/GPR54 (G-protein coupled receptor 54) system. METHODS: KiSS-1, GPR54, and ERalpha (estrogen receptor alpha) mRNA expression was quantified in control (C) and treated (T) maternal and fetal (110-day) hypothalami and pituitary glands using semiquantitative reverse transcription polymerase chain reaction, and colocalization of kisspeptin with LHbeta (luteinizing hormone beta) and ERalpha in C and T fetal pituitary glands quantified using dual-labeling immunohistochemistry. RESULTS: Fetuses exposed in utero to the EDC mixture showed reduced KiSS-1 mRNA expression across three hypothalamic regions examined (rostral, mid, and caudal) and had fewer kisspetin immunopositive cells colocalized with both LHbeta and ERalpha in the pituitary gland. In contrast, treatment had no effect on parameters measured in the adult ewe hypothalamus or pituitary. CONCLUSIONS: This study demonstrates that the developing fetus is sensitive to real-world mixtures of environmental chemicals, which cause significant neuroendocrine alterations. The important role of kisspeptin/GPR54 in regulating puberty and adult reproduction means that in utero disruption of this system is likely to have long-term consequences in adulthood and represents a novel, additional pathway through which environmental chemicals perturb human reproduction. PMID- 20019907 TI - Evidence for dose-additive effects of pyrethroids on motor activity in rats. AB - BACKGROUND: Pyrethroids are neurotoxic insecticides used in a variety of indoor and outdoor applications. Previous research characterized the acute dose-effect functions for 11 pyrethroids administered orally in corn oil (1 mL/kg) based on assessment of motor activity. OBJECTIVES: We used a mixture of these 11 pyrethroids and the same testing paradigm used in single-compound assays to test the hypothesis that cumulative neurotoxic effects of pyrethroid mixtures can be predicted using the default dose-addition theory. METHODS: Mixing ratios of the 11 pyrethroids in the tested mixture were based on the ED30 (effective dose that produces a 30% decrease in response) of the individual chemical (i.e., the mixture comprised equipotent amounts of each pyrethroid). The highest concentration of each individual chemical in the mixture was less than the threshold for inducing behavioral effects. Adult male rats received acute oral exposure to corn oil (control) or dilutions of the stock mixture solution. The mixture of 11 pyrethroids was administered either simultaneously (2 hr before testing) or after a sequence based on times of peak effect for the individual chemicals (4, 2, and 1 hr before testing). A threshold additivity model was fit to the single-chemical data to predict the theoretical dose-effect relationship for the mixture under the assumption of dose additivity. RESULTS: When subthreshold doses of individual chemicals were combined in the mixtures, we found significant dose-related decreases in motor activity. Further, we found no departure from the predicted dose-additive curve regardless of the mixture dosing protocol used. CONCLUSION: In this article we present the first in vivo evidence on pyrethroid cumulative effects supporting the default assumption of dose addition. PMID- 20019908 TI - Model development and validation of personal exposure to volatile organic compound concentrations. AB - BACKGROUND: Direct measurement of exposure to volatile organic compounds (VOCs) via personal monitoring is the most accurate exposure assessment method available. However, its wide-scale application to evaluating exposures at the population level is prohibitive in terms of both cost and time. Consequently, indirect measurements via a combination of microenvironment concentrations and personal activity diaries represent a potentially useful alternative. OBJECTIVE: The aim of this study was to optimize a model of personal exposures (PEs) based on microenvironment concentrations and time/activity diaries and to compare modeled with measured exposures in an independent data set. MATERIALS: VOC PEs and a range of microenvironment concentrations were collected with active samplers and sorbent tubes. Data were supplemented with information collected through questionnaires. Seven models were tested to predict PE to VOCs in 75% (n = 370) of the measured PE data set, whereas the other 25% (n = 120) was used for validation purposes. RESULTS: The best model able to predict PE with independence of measurements was based upon stratified microenvironment concentrations, lifestyle factors, and individual-level activities. The proposed model accounts for 40-85% of the variance for individual VOCs and was validated for almost all VOCs, showing normalized mean bias and mean fractional bias below 25% and predicting 60% of the values within a factor of 2. CONCLUSIONS: The models proposed identify the most important non-weather-related variables for VOC exposures; highlight the effect of personal activities, use of solvents, and exposure to environmental tobacco smoke on PE levels; and may assist in the development of specific models for other locations. PMID- 20019909 TI - Postnatal cadmium exposure, neurodevelopment, and blood pressure in children at 2, 5, and 7 years of age. AB - BACKGROUND: Adverse health effects of cadmium in adults are well documented, but little is known about the neuropsychological effects of cadmium in children, and no studies of cadmium and blood pressure in children have been conducted. OBJECTIVE: We examined the potential effects of low-level cadmium exposure on intelligence quotient, neuropsychological functions, behavior, and blood pressure among children, using blood cadmium as a measure of exposure. METHODS: We used the data from a multicenter randomized clinical trial of lead-exposed children and analyzed blood cadmium concentrations using the whole blood samples collected when children were 2 years of age. We compared neuropsychological and behavioral scores at 2, 5, and 7 years of age by cadmium level and analyzed the relationship between blood cadmium levels at 2 years of age and systolic and diastolic blood pressure at 2, 5, and 7 years of age. RESULTS: The average cadmium concentration of these children was 0.21 microg/L, lower than for adults in the National Health and Nutrition Examination Survey (NHANES), but comparable to concentrations in children < 3 years of age in NHANES. Except for the California Verbal Learning Test for Children, there were no differences in test scores among children in different cadmium categories. For children with detectable pretreatment blood cadmium, after adjusting for a variety of covariates, general linear model analyses showed that at none of the three age points was the coefficient of cadmium on Mental Development Index or IQ statistically significant. Spline regression analysis suggested that behavioral problem scores at 5 and 7 years of age tended to increase with increasing blood cadmium, but the trend was not significant. We found no significant associations between blood cadmium levels and blood pressure. CONCLUSION: We found no significant associations between background blood cadmium levels at 2 years of age and neurodevelopmental end points and blood pressure at 2, 5, and 7 years of age. The neuropsychological or hypertensive effects from longer background exposures to cadmium need further study. PMID- 20019910 TI - Urinary phthalate metabolites in relation to preterm birth in Mexico city. AB - BACKGROUND: Rates of preterm birth have been rising over the past several decades. Factors contributing to this trend remain largely unclear, and exposure to environmental contaminants may play a role. OBJECTIVE: We investigated the relationship between phthalate exposure and preterm birth. METHODS: Within a large Mexican birth cohort study, we compared third-trimester urinary phthalate metabolite concentrations in 30 women who delivered preterm (< 37 weeks of gestation) with those of 30 controls (> or = 37 weeks of gestation). RESULTS: Concentrations of most of the metabolites were similar to those reported among U.S. females, although in the present study mono-n-butyl phthalate (MBP) concentrations were higher and monobenzyl phthalate (MBzP) concentrations lower. In a crude comparison before correcting for urinary dilution, geometric mean urinary concentrations were higher for the phthalate metabolites MBP, MBzP, mono(3-carboxylpropyl) phthalate, and four metabolites of di(2-ethyl-hexyl) phthalate among women who subsequently delivered preterm. These differences remained, but were somewhat lessened, after correction by specific gravity or creatinine. In multivariate logistic regression analysis adjusted for potential confounders, elevated odds of having phthalate metabolite concentrations above the median level were found. CONCLUSIONS: We found that phthalate exposure is prevalent among this group of pregnant women in Mexico and that some phthalates may be associated with preterm birth. PMID- 20019911 TI - Predictors of serum dioxins and PCBs among peripubertal Russian boys. AB - BACKGROUND: Although sources and routes of exposure to dioxins and polychlorinated biphenyls (PCBs) have been studied, information regarding exposure among children is limited. Breast-feeding and diet are two important contributors to early life exposure. To further understand other significant contributors to childhood exposure, we studied a cohort of children from a city with high environmental dioxin levels. OBJECTIVES: We investigated predictors of serum concentrations of polychlorinated dibenzo-p-dioxins (PCDDs)/polychlorinated dibenzofurans (PCDFs)/co-planar PCBs (C-PCBs), toxic equivalents (TEQs), and PCBs among 8- to 9-year-old boys in Chapaevsk, Russia. METHODS: We used general linear regression models to explore associations of log(10)-transformed serum concentrations of PCDDs/PCDFs/C-PCBs, TEQs, and PCBs at study entry with anthropometric, demographic, geographic, and dietary factors in 482 boys in Chapaevsk, Russia. RESULTS: The median (25th, 75th percentile) concentration for total 2005 TEQs was 21.1 pg/g lipid (14.4, 33.2). Boys who were older, consumed local foods, were breast-fed longer, and whose mothers were employed at the Khimprom chemical plant (where chlorinated chemicals were produced) or gardened locally had significantly higher serum dioxins and PCBs, whereas boys with higher body mass index or more educated parents had significantly lower serum dioxins and PCBs. Boys who lived < 2 km from Khimprom had higher total TEQs (picograms per gram lipid) [adjusted mean = 30.6; 95% confidence interval (CI), 26.8-35.0] than boys who lived > 5 km away (adjusted mean = 18.8; 95% CI, 17.2-20.6). CONCLUSIONS: Our findings suggest that there are specific local sources of dioxin and PCB exposure among children in Chapaevsk including maternal gardening, consumption of locally grown food, and residential proximity to the Khimprom plant. PMID- 20019912 TI - Exposure to hydroxylated polychlorinated biphenyls (OH-PCBs) in the prenatal period and subsequent neurodevelopment in eastern Slovakia. AB - BACKGROUND: Hydroxylated polychlorinated biphenyls (OH-PCBs), unlike PCBs, are in general readily excreted yet are still detected in humans and animals. Active transport of OH-PCBs across the placenta and hydroxylation of PCBs by the fetus suggest the potential for greater impact on the fetus compared with the parent PCB compounds, but little is known about their health effects, particularly in humans. OBJECTIVES: The objective of this study was to evaluate the associations between prenatal OH-PCB exposure and neurodevelopment in children at 16 months of age in eastern Slovakia. METHODS: A birth cohort (n = 1,134) was enrolled during 2002-2004. We analyzed six OH-PCB metabolites (4-OH-CB-107, 3-OH-CB-153, 4-OH-CB 146, 3'-OH-CB-138, 4-OH-CB-187, and 4'-OH-CB-172) in a subset of the cohort. The Bayley Scales of Infant Development were administered to the children at the 16 month follow-up visit. We developed multiple linear regression models predicting standardized scores for the Mental Development Index (MDI) and Psychomotor Development Index (PDI) from maternal (n = 147) and cord (n = 80) serum OH-PCB concentrations, adjusting for sex of child, district, HOME (Home Observation for Measurement of the Environment) score, and maternal score on Raven's Progressive Matrices. RESULTS: Cord 4-OH-CB-107 was significantly associated with lower MDI (beta = -2.27; p = 0.01) and PDI (beta = -4.50; p = 0.004). Also, maternal 4-OH CB-107 was significantly associated with lower MDI (beta = -1.76; p = 0.03) but not PDI. No other OH-PCB metabolites were associated with decreased PDI or MDI. CONCLUSIONS: Our findings showed a significant association of 4-OH-CB-107 with decreased MDI, which can possibly be mediated by endocrine disruption, altered neurotransmitter functions, or reduced thyroid hormone concentrations in brain. PMID- 20019913 TI - Lead exposure and behavior among young children in Chennai, India. AB - BACKGROUND: Lead exposure has long been associated with deficits in IQ among children. However, few studies have assessed the impact of lead on specific domains of behavior and cognition. OBJECTIVE: We evaluated the associations between lead and different domains of neurobehavior and their relative sensitivity to lead. METHODS: We determined blood lead levels using a LeadCare instrument in 756 children 3-7 years of age attending pre- and elementary schools in Chennai, India. Anxiety, social problems, inattention, hyperactivity, and attention deficit hyperactivity disorder (ADHD), as well as executive function were assessed in children by their schoolteachers using Conners' Teacher Rating Scales-39, Conners' ADHD/Diagnostic and Statistical Manual for Mental Disorders, 4th Edition Scales (CADS), and the Behavior Rating Inventory of Executive Function questionnaires, with higher scores denoting worse behavior. Analyses were carried out using multivariate generalized estimating equations with comparisons of outcome Z-scores to assess the relative strengths of the associations between log-blood lead and the different domains of behavior. RESULTS: Mean blood lead level was 11.4 +/- 5.3 microg/dL. Blood lead was associated with higher anxiety (beta = 0.27, p = 0.01), social problems (beta = 0.20, p = 0.02), and higher scores in the ADHD index (beta = 0.17; p = 0.05). The effect estimate was highest for global executive function (beta = 0.42; p< 0.001). CONCLUSIONS: Higher blood lead levels in this population of young children is associated with increased risk of neurobehavioral deficits and ADHD, with executive function and attention being particularly vulnerable domains to the effects of lead. PMID- 20019914 TI - Association of traffic-related air pollution with children's neurobehavioral functions in Quanzhou, China. AB - BACKGROUND: With the increase of motor vehicles, ambient air pollution related to traffic exhaust has become an important environmental issue in China. Because of their fast growth and development, children are more susceptible to ambient air pollution exposure. Many chemicals from traffic exhaust, such as carbon monoxide, nitrogen dioxide, and lead, have been reported to show adverse effects on neurobehavioral functions. Several studies in China have suggested that traffic exhaust might affect neurobehavioral functions of adults who have occupational traffic exhaust exposure. However, few data have been reported on the effects on neurobehavioral function in children. OBJECTIVES: The objective of this study was to explore the association between traffic-related air pollution exposure and its effects on neurobehavioral function in children. METHODS: This field study was conducted in Quanzhou, China, where two primary schools were chosen based on traffic density and monitoring data of ambient air pollutants. School A was located in a clear area and school B in a polluted area. We monitored NO(2) and particulate matter with aerodynamic diameter < or = 10 mum as indicators for traffic-related air pollution on the campuses and in classrooms for 2 consecutive days in May 2005. The children from second grade (8-9 years of age) and third grade (9-10 years of age) of the two schools (n = 928) participated in a questionnaire survey and manual-assisted neurobehavioral testing. We selected 282 third-grade children (school A, 136; school B, 146) to participate in computer assisted neurobehavioral testing. We conducted the fieldwork between May and June 2005. We used data from 861 participants (school A, 431; school B, 430) with manual neurobehavioral testing and from all participants with computerized testing for data analyses. RESULTS: Media concentrations of NO(2) in school A and school B campus were 7 microg/m(3) and 36 microg/m(3), respectively (p < 0.05). The ordinal logistic regression analyses showed that, after controlling the potential confounding factors, participants living in the polluted area showed poor performance on all testing; differences in results for six of nine tests (66.7%) achieved statistical significance: Visual Simple Reaction Time with preferred hand and with nonpreferred hand, Continuous Performance, Digit Symbol, Pursuit Aiming, and Sign Register. CONCLUSION: We found a significant relationship between chronic low-level traffic-related air pollution exposure and neurobehavioral function in exposed children. More studies are needed to explore the effects of traffic exhaust on neurobehavioral function and development. PMID- 20019915 TI - Drinking-water herbicide exposure in Indiana and prevalence of small-for gestational-age and preterm delivery. AB - BACKGROUND: Atrazine and other corn herbicides are routinely detected in drinking water. Two studies on potential association of atrazine with small-for gestational-age (SGA) and preterm birth prevalence found inconsistent results. Moreover, these studies did not control for individual-level potential confounders. OBJECTIVES: Our retrospective cohort study evaluated whether atrazine in drinking water is associated with increased prevalence of SGA and preterm birth. METHODS: We developed atrazine concentration time series for 19 water systems in Indiana from 1993 to 2007 and selected all births (n = 24,154) based on geocoded mother's residences. Log-binomial models were used to estimate prevalence ratios (PRs) for SGA and preterm delivery in relation to atrazine concentrations during various periods of the pregnancy. Models controlled for maternal demographic characteristics, prenatal care and reproductive history, and behavioral risk factors (smoking, drinking, drug use). RESULTS: Atrazine in drinking water during the third trimester and the entire pregnancy was associated with a significant increase in the prevalence of SGA. Atrazine in drinking water > 0.1 microg/L during the third trimester resulted in a 17-19% increase in the prevalence of SGA compared with the control group (< 0.1 microg/L). Mean atrazine concentrations over the entire pregnancy > 0.644 microg/L were associated with higher SGA prevalence than in the control group (adjusted PR = 1.14; 95% confidence interval, 1.03-1.24). No significant association was found for preterm delivery. CONCLUSIONS: We found that atrazine, and perhaps other co-occurring herbicides in drinking water, is associated with an increased prevalence of SGA, but not preterm delivery. PMID- 20019916 TI - Do human milk concentrations of persistent organic chemicals really decline during lactation? Chemical concentrations during lactation and milk/serum partitioning. AB - BACKGROUND: Conventional wisdom regarding exposures to persistent organic chemicals via breast-feeding assumes that concentrations decline over the course of lactation and that the mother's body burden reflects her cumulative lifetime exposure. Two important implications stemming from these lines of thought are, first, that assessments of early childhood exposures should incorporate decreasing breast milk concentrations over lactation; and, second, that there is little a breast-feeding mother can do to reduce her infant's exposures via breast feeding because of the cumulative nature of these chemicals. OBJECTIVES: We examined rates of elimination and milk/serum partition coefficients for several groups of persistent organic chemicals. METHODS: We collected simultaneous milk and blood samples of 10 women at two times postpartum and additional milk samples without matching blood samples. RESULTS: Contrary to earlier research, we found that lipid-adjusted concentrations of polybrominated diphenyl ethers, polychlorinated biphenyls, polychlorinated dibenzo-p-dioxins and furans, and organochlorine pesticides in serum and milk do not consistently decrease during lactation and can increase for some women. Published research has also suggested an approximate 1:1 milk/serum relationship (lipid adjusted) on a population basis for 2,3,7,8-tetrachlorodibenzo-p-dioxin; however, our results suggest a more complex relationship for persistent, lipophilic chemicals with the milk/serum relationship dependent on chemical class. CONCLUSIONS: Decreases in concentration of lipophilic chemicals on a lipid-adjusted basis during lactation should no longer be assumed. Thus, the concept of pumping and discarding early milk as means of reducing infant exposure is not supported. The hypothesis that persistent lipophilic chemicals, on a lipid-adjusted basis, have consistent concentrations across matrices is likely too simplistic. PMID- 20019918 TI - The future is now. PMID- 20019917 TI - Developmental changes in PON1 enzyme activity in young children and effects of PON1 polymorphisms. AB - BACKGROUND: Paraoxonase 1 (PON1) is an enzyme that detoxifies activated organophosphorus pesticides (OPs) and is also involved in oxidative stress pathways. OBJECTIVES: PON1 activity in newborns is lower than in adults, but the ontogeny of PON1 activity is poorly characterized in young children. We examined the effects of age and PON1 genotype on enzyme activity in a birth cohort of Mexican-American children. METHODS: We determined three substrate-specific measures of PON1 activity in 1,143 plasma samples collected longitudinally from 458 children at five time points from birth through 7 years of age, and genotyped PON1 polymorphisms at positions 192 and -108 in these children. RESULTS: Contrary to previous reports that PON1 activities plateau by 2 years of age, we observed an age-dependent increase in all three PON1 measures from birth through 7 years of age (p < 0.0001). The PON1(192) genotype significantly modified the effect of age on paraoxonase (POase) activity (p < 0.0001) such that increases in enzyme activity with age were influenced by the number of R alleles in a dose-dependent manner. Children with the PON1(-108CC192RR) diplotype had significantly higher mean PON1 activities and also experienced steeper increases of POase activity over time compared with children with the PON1(-108TT192QQ) diplotype. CONCLUSIONS: Lower levels of the PON1 enzyme, which is involved in protection against OPs and oxidative stress, persist in young children past 2 years of age through at least 7 years of age. Future policies addressing pesticide exposure in children should take into account that the window of vulnerability to OPs in young children may last beyond infancy. PMID- 20019919 TI - An ethics-based approach to global child health research. PMID- 20019920 TI - Adolescent health - A developing international discipline. PMID- 20019921 TI - Management of fever without source in children: Changing times. PMID- 20019922 TI - Approach to the febrile child: A challenge bridging the gap between the literature and clinical practice. AB - The approach to the febrile child is always concerning for any physician despite the fact that most fevers are viral in origin. However, in rare cases, a missed bacterial infection can have serious consequences. How can fevers of viral origin be differentiated from those of bacterial origin? Do all febrile children with no obvious infection site need a blood culture? Should antibiotics be administered before the results of the blood culture have been received? In the past 30 years, there has been an overabundance of recommendations, advice, opinions and suggested treatments on this subject. The purpose of this review is to present the evidence that is known at this time concerning the management of the febrile child and to present one approach used in a large urban paediatric emergency department. PMID- 20019923 TI - The presence of clinical signs in malnourished infants with acute lower respiratory tract infections. AB - OBJECTIVES: To determine the reliability of respiratory rate and subcostal retractions in diagnosing acute lower respiratory tract infection in malnourished children. METHODS: One hundred forty-three children with acute lower respiratory tract infection were classified according to the Gomez classification as normal, mildly, moderately or severely malnourished. The presence of tachypnea, subcostal retractions and the sensitivity of either sign in identifying children with a clinical and radiological diagnosis of acute lower respiratory tract infection in each of the nutritional categories were evaluated and compared. RESULTS: According to the Gomez classification, 21 (15%) of 143 subjects were severely malnourished, 40 (28%) were moderately malnourished, 38 (26%) were mildly malnourished and 44 (31%) were well nourished. The mean respiratory rates in subjects with normal nutrition and in mildly, moderately and severely malnourished subjects were 62.6+/-9.38 breaths/min, 61.3+/-5.57 breaths/min, 57.6+/-11.65 breaths/min and 49.9+/-9.04 breaths/min, respectively. The mean respiratory rate of those with normal nutrition was not significantly different from that of those with mild malnutrition (P>0.05), but there was a statistically significant difference between the normal subjects and both the moderately malnourished (P=0.03) and severely malnourished (P<0.001) subjects. Subcostal retraction was observed in 78% of total children, which represents 41 (93%) subjects with normal nutrition, 34 (89%) mildly malnourished subjects, 30 (75%) moderately malnourished subjects and 10 (47%) severely malnourished subjects. There was no statistically significant difference between the frequencies of subcostal retractions observed in children with normal nutrition and mildly malnourished children (P>0.05), but subcostal retraction frequencies were significantly lower both in moderately (P=0.03) and severely (P<0.001) malnourished children. Tachypnea frequency was also found to be significantly lower in moderately (P=0.04) and severely (P=0.03) malnourished children compared with the normal group. As a consequence, the sensitivity of the presence of both of these signs was found to be lower in these two groups. The coexistence rates of both findings were 95% for the patients with normal nutrition and 47% for the severely malnourished patients. CONCLUSIONS: The current World Health Organization algorithm must be supported with other signs for the diagnosis of acute lower respiratory tract infection, especially in severely mal-nourished children. PMID- 20019924 TI - Home fortification with micronutrient sprinkles - A new approach for the prevention and treatment of nutritional anemias. AB - Despite global goals set by United Nations' agencies over the past decade for significant reductions in iron deficiency anemia (IDA), it remains a largely unaddressed public health problem affecting more than two billion people, one third of the world's population. The negative impact of IDA on health and human potential are greatest in the developing world, where it is estimated that 51% of children younger than four years of age are anemic, mainly due to a diet that is inadequate in bioavailable iron. Studies in both developed and developing countries have consistently shown mental and motor impairments that may not be reversible in children younger than two years of age with IDA. From a public health standpoint there are four possible interventions for the prevention of anemia: dietary diversification to include foods rich in absorbable iron; fortification of staple foods including targeted fortification of complementary foods for infants and young children; the provision of iron supplements; and 'home-fortification'. In response to a United Nations Children's Fund (UNICEF) request to develop a new approach to IDA, our research group developed 'Sprinkles' for home-fortification of complementary foods. Sprinkles are single dose sachets (like small packets of sugar) containing micronutrients in powder form (encapsulated iron, zinc, vitamins A, C and D, and folic acid), which are easily sprinkled onto any home-prepared complementary food. Sprinkles were developed to overcome many of the side effects and disadvantages of iron drops. We have demonstrated that Sprinkles are as effective as iron drops in the treatment and prevention of anemia. Sprinkles are easier to use and are, therefore, better accepted than iron drops, which may improve adherence to iron interventions. PMID- 20019925 TI - Bioterrorism - Health emergency preparedness and response. AB - Health emergency planning for preparedness and response against acts of terrorism, including the malfeasant threat or actual release of biological agents designed to harm others, has assumed a higher level of concern for most western nations, including Canada, following the explosive attacks in the United States on September 11, 2001. These terrorist attacks were followed by an outbreak of anthrax infections. The Bacillus anthracis spores in these attacks were dispersed by using regular postal services in the United States. In addition to the unsettling sense of social vulnerability that resulted from these attacks, a greater appreciation that the integration of public health, emergency health and social services with security activities was required to fully address the need to protect the health and other interests of the citizens. Collaborative work among regional, provincial, territorial, federal and international authorities within these domains is emerging as an effective response to the risk management of bioterrorism. The following is a brief description of the health framework for preparedness and response, and the biological agents of major concern in terrorism. PMID- 20019927 TI - Cretinism: The past, present and future of diagnosis and cure. PMID- 20019926 TI - Travellers' diarrhea in children. AB - Diarrhea is the most common medical problem affecting all travellers to developing countries. Younger children are at especially high risk of acquiring travellers' diarrhea and of suffering more severe consequences. Up to 50% of travellers from developed to developing countries can expect to have at least one episode of acute diarrhea during a two-week stay. Episodes of travellers' diarrhea usually begin abruptly, either during travel or soon after returning home, and are generally self-limited. PMID- 20019929 TI - Clinician's Corner. PMID- 20019928 TI - Congenital rubella syndrome - Time to act on missed prevention opportunities. PMID- 20019930 TI - Clinician's Corner. PMID- 20019931 TI - Swimming lessons for infants and toddlers. PMID- 20019932 TI - Swimming and water safety for young children. PMID- 20019933 TI - What the paediatrician should know about paediatric and adolescent gynecology: The perspective of a gynecologist. AB - Gynecological problems are common in children but training in this subject is rare during paediatric postgraduate education. Knowledge in this area is important for providing comprehensive health care to children. The article explores the gynecological examination of the child and common gynecological disorders in the paediatric and adolescent age groups. Routine examination of the external genitalia as part of a well-child physical normalizes the examination for the child and helps to inform the physician about the large variations in normal genitals. Many complaints can be dealt with by reassurance, while most others respond to medical therapy and can be dealt with easily by paediatricians. Both young children and adolescents respond positively when allowed to have control over the tempo of the interview and the examination. A positive experience associated with a genital examination will help to open lines of trust. This along with nonjudgmental, open communication, will create educational opportunities to enhance a responsible transition from child to young adult. PMID- 20019934 TI - Orthotopic liver transplantation in a patient with carbamyl phosphate synthetase deficiency and cystic fibrosis. AB - A 15-year-old female with carbamyl phosphate synthetase deficiency, cystic fibrosis, and cystic fibrosis-related diabetes underwent orthotopic cadaveric liver transplantation. Metabolic control was maintained during the procedure with nutritional support and the use of intravenous sodium phenylacetate and benzoate. Her postoperative course was complicated by seizures and a transient decline in her pulmonary function tests, which returned to preoperative levels within one year of the transplant. Now, four years post-transplant, her quality of life has dramatically improved. There are only four Canadian centres with paediatric liver transplantation programs. However, expert medical care for adults with inborn error of metabolism is even more limited, suggesting that access to adult medical care is one of the many factors to be considered when liver transplantation is contemplated for patients with metabolically unstable conditions. PMID- 20019935 TI - Joubert syndrome: Report of a neonatal case. AB - Joubert syndrome is an autosomal recessive disorder that is characterized by a variable combination of central nervous system, respiratory and eye anomalies. It is a syndrome with a variable phenotype: partial or complete absence of the cerebellar vermis is seen in all patients, while other cardinal findings include episodic tachypnea and apnea in the neonatal period, jerky eye movements, hypotonia, severe mental handicap, developmental delay, ataxia and impaired equilibrium. Even within sibships the phenotype may vary, making it difficult to establish the exact clinical diagnostic boundaries of Joubert syndrome. A case of Joubert syndrome in a newborn is reported and the importance of recognizing the syndrome in the neonatal period so that specific and effective supportive measures can be started as soon as possible is stressed. PMID- 20019936 TI - Procedural sedation and analgesia for paediatric patients in the emergency department. AB - Children presenting to the emergency department (ED) often require sedation for brief procedures such as fracture and dislocation reductions, laceration repairs, and imaging procedures that are painful, anxiety provoking or both. This article presents three cases of paediatric patients who require sedation and/or analgesia, and summarizes important aspects of procedural sedation for the primary care practitioner in the emergency setting. Presedation assessment and monitoring equipment are detailed. Discussion of routes of administration and different agents including barbiturates, opiates, benzodiaxepines, the 'cardiac coctail', ketamine, propofol, nitrous oxide, and etomidate follow. Emphasis is placed on indications, contraindications, dosing, timing and advantages and disadvantages of each. Reversal agents are mentioned, and discharge criteria are outlined. PMID- 20019937 TI - Lead poisoning. PMID- 20019938 TI - Helping to prevent obesity and complications in children with Prader-Willi syndrome. PMID- 20019939 TI - Use of propofol sedation in the paediatric emergency department. PMID- 20019940 TI - Haemophilus influenzae type b cases in 2001-2002. PMID- 20019942 TI - Pacifiers (soothers): A user's guide for parents. PMID- 20019941 TI - Recommendations for the use of pacifiers. PMID- 20019944 TI - Ophthalmic emergencies in infants and children. PMID- 20019943 TI - Use of whole cow's milk in late infancy - Change tack or stay the course? PMID- 20019945 TI - When should waiting time become a quality of care issue in the paediatric emergency department? PMID- 20019946 TI - Quality of after-hours care. PMID- 20019947 TI - Whole cow's milk in infancy. AB - Early introduction of whole cow's milk may lead to iron deficiency anemia. From a nutritional point of view, it is best to delay the introduction of whole cow's milk until the infant is one year old. While there is no evidence to suggest adverse clinical sequelae associated with the increased renal solute load in healthy infants, feeding with whole cow's milk would narrow the margin of safety in situations that may lead to dehydration. Early exposure to cow's milk proteins increases the risk of developing allergy to milk proteins. Because of the possible association between early exposure to cow's milk proteins and risk for type 1 diabetes mellitus, breast-feeding and avoidance of commercially available cow's milk and products containing intact cow's milk protein during the first year of life are strongly encouraged in families with a strong history of insulin dependent diabetes mellitus. The authors suggest that the optimal food in infancy is human breast milk. If human milk is not available, it is preferred that iron fortified formulas rather than whole cow's milk be used during the first year of life. PMID- 20019948 TI - Pagers in a busy paediatric emergency waiting room: A randomized controlled trial. AB - OBJECTIVE: To test whether the distribution of pagers to waiting families in a paediatric emergency department (ED) increases satisfaction with the visit. METHODS: Using a controlled, randomized, single blind design, English-speaking families who had waited longer than 1 h were enrolled in the study. On any one day, every enrolled family was assigned to a no treatment group, an active control group in which children received a colouring set or an experimental group in which families were given pagers. A telephone questionnaire was used to measure the degree of satisfaction. Additional questions ascertained satisfaction with the pager and gathered other information. RESULTS: There were 23 families in the no treatment group, 18 in the active control group, and 20 in the experimental group. Demographic data were similar among the groups. Although satisfaction was higher in the experimental group, the differences between groups were not significant. The sex of the child or accompanying adult, waiting times, or classification as trauma or nontrauma did not influence the results. Families said pagers were useful (mean score 1.45 [1 = strongly agree]) and thought that the ED should use them (mean score 1.3). More families in the pager group left the waiting room while waiting to be seen (16 of 20 versus 19 of 40, P=0.016). The average score for finding the intervention helpful was 1.5 in the pager group and 3.06 in the colouring book group (P=0.0002). CONCLUSIONS: Use of a pager in the ED was viewed favourably by parents. However, overall satisfaction may be influenced more strongly by factors other than how the waiting time is spent. PMID- 20019949 TI - Does a paediatric after-hours clinic use evidence-based guidelines in the management of acute otitis media? AB - OBJECTIVE: To determine whether a paediatric after-hours clinic uses evidence based management in the treatment of acute otitis media, and compare this management with that provided in a paediatric emergency department and a general hospital emergency department. METHODS: A retrospective chart review of 573 patients (aged six months to five years) with a discharge diagnosis of acute otitis media was conducted in three after-hours settings: a paediatric after hours clinic, a tertiary paediatric hospital emergency department and a secondary general hospital emergency department. The patients' age, weight, sex and allergy to antibiotics were recorded as baseline characteristics. The physicians' antibiotic choice, dose and duration, and the use of investigations were recorded as outcome variables. RESULTS: Amoxicillin was prescribed to 68% of patients at both the paediatric after-hours clinic and the paediatric hospital emergency department, compared with 53% of patients at the general hospital emergency department (P<0.01). The mean dose of amoxicillin prescribed at the paediatric after-hours clinic and the paediatric hospital emergency department were similar (43.4+/-9.7 mg/kg per day and 42.4+/-14.3 mg/kg per day, respectively) and higher than that prescribed at the general hospital emergency department (38.6+/-8.8 mg/kg per day, P<0.01). The paediatric after-hours clinic used investigations less often than did emergency departments (0.5% of cases compared with 9% and 20%, P<0.01). CONCLUSION: The paediatric after-hours clinic provided a high level of adherence to a clinical practice guideline and had a low utilization of resource intensive investigations. PMID- 20019950 TI - Home injury patterns in children: A comparison by hospital sites. AB - BACKGROUND: Many intervention studies typically require data from several centres to ensure adequate power. The usual intention is to pool data after testing for heterogeneity. Sites that differ in sample characteristics may, on the one hand, complicate the assessment of the intervention, but on the other hand, they may add important insights through analysis of site-specific findings. OBJECTIVES: The aims of the present paper were to compare the distribution of injuries and risk factors among children participating in a five-centre study of a home-based injury prevention program, and to contrast parental injury awareness and knowledge with home safety measures. METHODS: Five children's hospitals in Canada agreed to participate in a case-control study combined with a randomized controlled trial. Patients were children zero to seven years of age presenting to a hospital emergency department with a fall, burn, ingestion or choking. Two controls were matched to each case, one with another injury and another with a minor illness. A home visitor completed a home hazard assessment based on observed safety measures. To determine whether data could be pooled, comparisons across sites were made with respect to types of injuries seen, sociodemographic characteristics, observed hazards and the parents' reported beliefs about severity of injuries, safety measures, preventability of injuries and susceptibility to injuries. RESULTS: There were few differences between the five hospitals. The mean age was 2.2 years (range 1.4 to 3.3). There were 219 falls (56%), 80 burns (20.4%), 54 poisonings (13.8%), and 38 chokings (9.7%), all distributed in a proportionately similar manner, except for poisoning, at each site. There were significantly more well-educated fathers at one hospital and younger parents with less education at another. Homes were generally lacking five recommended safety measures. However, most parents at all sites perceived their home as being very safe for any of the specific injuries, and their child as being at low risk of sustaining any of these injuries. CONCLUSIONS: The similarity across sites supports the pooling of these data regarding hospital treated injuries in young children in urban Canada. Most parents at all sites perceived their home as being very safe in spite of their homes lacking one quarter of the recommended safety measures. This discrepancy between parental perception and home safety highlights the needs for further education and prevention efforts. PMID- 20019951 TI - Fatal late onset group B streptococcal meningitis following maternal postpartum sepsis. AB - Although maternal screening and the administration of prophylactic intrapartum antibiotics have decreased the incidence of early onset group B streptococcal (GBS) disease in neonates, there is still significant morbidity and mortality as a result of neonatal GBS disease.Maternal GBS infections are not uncommon, but with appropriate therapy there is almost a uniformly good outcome. Little is written about the appropriate management of well infants born to mothers with postpartum GBS sepsis.The question of whether well infants born to mothers with GBS puerperal sepsis should be treated empirically with antibiotics and the lack of literature concerning this issue became apparent when an untreated term infant died of late onset GBS meningitis following maternal puerperal GBS sepsis. We describe this event in the following case presentation.With the current paucity of literature regarding the management of well infants born to mothers with postpartum GBS sepsis, it seems prudent to treat such infants empirically with antibiotics (following a full septic work-up) until this matter has been investigated further. PMID- 20019952 TI - In children with asthma who are currently using inhaled corticosteroids, are antileukotrienes more effective than placebo in improving clinical outcomes?: Part A. PMID- 20019953 TI - CPSP 2002 Results: What have we learned? PMID- 20019954 TI - Dietary manipulations for infantile colic. PMID- 20019956 TI - Dehydration and diarrhea. PMID- 20019955 TI - Treatment of diarrheal disease. PMID- 20019958 TI - Primary intratesticular leiomyosarcoma. AB - This report presents a case of primary intratesticular leiomyosarcoma. A 73-year old male presented with a 6-year history of left scrotal swelling. A radiological examination revealed a left testicular tumour with multiple metastases in the lung, para-aortic lymph node and other organs. A radical orchiectomy was carried out and the pathology revealed an intratesticular leiomyosarcoma. The patient received additional chemotherapy. Cases of primary intratesticular leiomyosarcoma are rare. This is, to the best of our knowledge, only the tenth case of leiomyisaocoma in an adult reported in the literature, and the first case involving multiple metastases. PMID- 20019959 TI - Urolithiasis in adults with congenital megaureter. AB - The primary presentation of congenital megaureter in adults is rare. Development of urolithiasis may lead to this unusual underlying diagnosis. Urinary tract stones can form either within the dilated ureteral segment or in a part of the upper urinary tract proximal to the abnormal ureteral segment. We report two cases of nephrolithiasis that occurred in adults found to have segmental megaureter. The first case is that of a 58-year-old man who presented with left lower quadrant pain. Computed tomography scan revealed a 2-cm stone in the distal left ureter within an area of isolated segmental distal ureteral dilation. The second case is a 48-year-old man who developed recurrent renal urolithiasis associated with isolated distal megaureter.Although a rare condition in adults, congenital megaureter may present when kidney stones develop as a result of the ureteral abnormality. Typically, stones will develop within the dilated segment of ureter. Atypically, stones may develop away from the site of the underlying abnormality. Congenital megaureter is a diagnosis that urologists and radiologists need to consider in the setting of isolated distal ureteral dilation, as the diagnosis of adult megaureter may require more involved surgical measures to prevent recurrence of adverse symptoms. PMID- 20019960 TI - Symptomatic cystic seminal vesicle: a laparoscopic approach for effective treatment. AB - This case report highlights a symptomatic cystic seminal vesicle lesion, treated laparoscopically. A young adult male known to have congenital right renal agenesis presented with a history of recurrent right iliac fossa pain, as well as deep pelvic discomfort. The preoperative evaluation revealed a 7 x 4.5 x 4 cm cystic seminal vesicle mass. After discovery of the mass, the patient underwent a transperitoneal laparoscopic excision of the lesion. The patient was discharged on the third postoperative day and had an uneventful recovery. There were no complications noted during a routine follow-up. The laparoscopic approach was shown to be effective in the management of this rare condition. PMID- 20019961 TI - Ureteral stent-related aortoureteric fistula: case report and literature review. AB - We describe a case of early postoperative aortoureteric fistula following exenterative surgery treated with wet ostomy with bilateral ureterosigmoidostomies and ureteric stent placement. A high index of clinical suspicion is required, as diagnosis is often difficult. PMID- 20019962 TI - Bilobed testicle in children: diagnosis and management. AB - Bilobed testicle is an extremely rare testicular malformation in children. It may be presumed that this condition could be an incomplete expression of polyorchidism; however, the etiology of both entities remains unknown. In this report, a 3-year-old boy presented with a right bilobed testicle mimicking a testicular tumour. Since there are insufficient data in the pediatric literature presenting the ideal management of bilobed testicles, we believe it is important to report this case to provide more information about this condition. Although rare, bilobed testis should be considered in the differential diagnosis of a testicular mass. Surgical treatment is not mandatory, and conservative follow-up may play a role. PMID- 20019963 TI - Malignant giant pheochromocytoma: a case report and review of the literature. AB - Malignant pheochromocytoma is a rare disease and surgical resection is the only curative treatment. There are no definitive histological or cytological criteria of malignancy, as it is impossible to determine this condition in the absence of advanced locoregional disease or metastases. We report a case of a patient with a giant retroperitoneal tumour, the second largest to be published, which was diagnosed as a malignant pheochromocytoma; it was treated with surgery. The literature is reviewed to evaluate tumour features and criteria to distinguish between benign and malignant pheochromocytomas. PMID- 20019964 TI - Dose escalation of second-line sunitinib results in rapid partial remission of multiple hepatic metastases. AB - A 58-year-old man with metastatic clear cell renal cell carcinoma on sunitinib therapy, who previously failed on sorafenib, was found to have progression of multiple hepatic metastases; he was on a standard sunitinib dose of 50 mg/day (4 weeks on, 2 weeks off). Due to the unavailability of alternative therapies, a sunitinib dose escalation of 50 mg/day was attempted. After one 6-week cycle of continuously dosed sunitinib 50 mg, the hepatic lesions regressed. After the second cycle, virtual disappearance of the lesions was recorded. There was no added toxicity. These findings suggest that sunitinib dose escalation to 50 mg/day using continuous daily administration dosing might represent a valid, effective and well-tolerated therapeutic option in patients who progress on standard sunitinib therapy. PMID- 20019965 TI - Mantle cell lymphoma of the prostate. AB - Lymphoma of the prostate, either primary or secondary, is very rare. We report the case of an 82-year-old man with symptoms of bladder outlet obstruction presumably due to benign prostate hyperplasia (BPH). He underwent an uneventful transurethral resection of the prostate. Pathological review of the resected tissue demonstrated substantial infiltration by an atypical lymphoid infiltrate positive for CD20, BCL2, CD5, kappa light chain and cycline D1. Histology and immunoprofile were consistent with mantle cell lymphoma. PMID- 20019966 TI - The importance of quality indicators: a call to action. PMID- 20019967 TI - Introduction: Key issues in bladder cancer management. PMID- 20019968 TI - CUA as health advocate. PMID- 20019969 TI - Changes in health utilities and health-related quality of life over 12 months following radical prostatectomy. AB - INTRODUCTION: Although radical prostatectomy (RP) is an effective treatment for prostate cancer, it has potentially deleterious effects on health-related quality of life (HRQoL). Utility is an important global measure of HRQoL. This utility is also used in decision-making models and economic evaluations. There are no published prospective data characterizing men's utilities for health outcomes post-RP. METHODS: From July 2003 to June 2006, patients undergoing RP at the University Health Network in Toronto, Ontario, Canada, completed the Patient Oriented Prostate Utility Scale (PORPUS), a disease-specific HRQoL and utility instrument. Men were included if they had clinically localized prostate cancer and had completed HRQoL measures prior to surgery and at least once within 9 to 18 months after the RP. Health-related quality of life was measured prior to surgery and at 4 stages after surgery: 0 to 3 months, 3 to 9 months, 9 to 18 months, and 18 to 30 months. Statistical analyses included pairwise t-tests and multivariable linear regression comparing changes in scores between baseline and the 9- to 18-month range. RESULTS: Two hundred and thirteen men (mean age 60.9, median Gleason score 7) met the inclusion criteria. At baseline, HRQoL was high (mean PORPUS-P [psychometric] = 83.8). Scores declined sharply by 3 months (65.5, p < 0.001) and improved but did not reach baseline by 9-18 months (75.1, p < 0.001). Utility scores (PORPUS-U[utility]) showed the same pattern: 0.94 at baseline; 0.81 at 3 months (p < 0.001); and 0.88 at 9 to 18 months (p < 0.001). Predictors of HRQoL, including baseline PORPUS score, nerve-sparing surgery and smoking status, decline after RP. CONCLUSION: Symptom scores are known to be affected up to 12 months after RP. Our study demonstrates that global health status also declines and remains affected 1 year after surgery. PMID- 20019970 TI - Measuring quality of life after radical prostatectomy from the patient's perspective. PMID- 20019971 TI - NMP22 is predictive of recurrence in high-risk superficial bladder cancer patients. AB - INTRODUCTION: The nuclear matrix protein 22 (NMP22) assay has been shown to have greater sensitivity for the diagnosis and detection of recurrent urothelial carcinoma of the bladder (UCB) over that of traditional urine cytology. We assessed the use of NMP22 to predict which high-risk superficial UCB patients will have recurrence, progression or disease-related death; we compared these results to standard urine cytology. METHODS: One hundred consecutive patients with high-risk superficial UCB were enrolled. During surveillance, urine was collected for cytology and NMP22 testing. Patients were followed for at least 6 months. Retrospective chart review was undertaken to collect data on previous tumour history, tumour characteristics, disease recurrences, progression and death. Kaplan-Meier analyses were performed to determine the significance between NMP22-positive and -negative patients in terms of recurrence-free, progression free and overall survival. Similar analyses were performed for urine cytology. RESULTS: From 94 eligible patients, 15 and 79 were NMP22 positive and negative, respectively. The baseline characteristics between the 2 groups were not significantly different in terms of patient characteristics, prior tumour history or intravesical therapies received. Mean recurrence-free survival time was significantly lower in the NMP22 positive group (p = 0.038); however, mean progression-free and overall survival were not significantly different between the 2 groups (p = 0.297 and 0.519, respectively). Urine cytology demonstrated no significant predictive power for disease recurrence, progression or survival. CONCLUSION: The nuclear matrix protein 22 assay appears to have predictive value for future tumour recurrences, but not progression or overall survival in patients with high-risk superficial UCB. PMID- 20019972 TI - NMP22: Does it have a predictive potential for recurrence and progression in non muscle-invasive bladder cancer? PMID- 20019973 TI - Hormone use after radiotherapy failure: a survey of Canadian uro-oncology specialists. AB - INTRODUCTION: A survey of Canadian uro-oncology specialists was performed to assess practice patterns of patients with recurrent prostate cancer postradiotherapy and to assess the feasibility of conducting a trial in this setting. METHODS: There were 14 survey questions and 1 demographic question. Responses were reported by frequency. RESULTS: There were 96 respondents. Most respondents use both prostate-specific antigen doubling time (PSAdt) and PSA level when deciding to start androgen deprivation therapy (ADT) in asymptomatic patients. About half of respondents start ADT when PSA is greater than 10 ng/mL or when the PSAdt is less than 6 months. Eighty-six percent felt that the timing of ADT was an important research question. Over 1500 patients per year were estimated as being available for such a trial. CONCLUSION: After radiotherapy failure, respondents initiated ADT about half of the time when PSA is less than 10 ng/mL and/or PSAdt is less than 6 months. A clinical trial examining the timing of ADT has strong support and appears to be feasible. PMID- 20019974 TI - The effect of nerve-sparing surgery on patient-reported continence post-radical prostatectomy. AB - INTRODUCTION: Urinary continence significantly affects quality of life after radical prostatectomy (RP). The impact of nerve-sparing surgery on continence is unclear from the current literature. METHODS: We identified men with prostate cancer from the University Health Network Prostate Centre database who underwent RP. Preoperatively and at each postoperative visit, patients completed the Patient-Oriented Prostate Utility Scale (PORPUS), a validated psychometric and health utility instrument. Incontinence was defined by a single questionnaire item. Patients with radiotherapy or less than 10 months follow-up were excluded. Chi-squared tests and ANOVA were used to compare groups. Multivariable logistic regression was used to control for effects of nerve-sparing and other covariates. RESULTS: Of the 253 eligible patients from 2003 to 2007, 159 patients had bilateral nerve-sparing, 32 had unilateral nerve-sparing and 62 had non-nerve sparing surgery. Of these patients, 27%, 17% and 34%, respectively, were classified as incontinent at 1 year. These proportions were not significantly different between groups (p = 0.23). Multivariable logistic regression showed baseline urinary continence and urinary frequency to be significant predictors of patient-reported continence at 1 year postoperatively, with odds ratios of 1.7 (95% confidence interval [CI] 1.1-2.9) and 1.5 (95% CI 1.0-2.3), respectively. There was a significant difference in the proportion of PORPUS sexual function scores between nerve-sparing groups after excluding those with baseline sexual dysfunction (p = 0.003). Similarly, health-related utility scores were different across groups (p < 0.001). CONCLUSION: Our results do not suggest a difference in 1-year patient-reported continence based on the type of nerve-sparing RP. However, baseline continence and urinary frequency were significant predictors of continence at 1 year. PMID- 20019975 TI - Improving outcomes post-radical prostatectomy: nerve-sparing status and urinary continence. PMID- 20019977 TI - Hunner's lesions. PMID- 20019976 TI - Endoscopic ablation of Hunner's lesions in interstitial cystitis patients. AB - INTRODUCTION: We report our experience with endoscopic ablation of Hunner's lesions in women with interstitial cystitis (IC). METHODS: A chart review was performed on 14 patients with IC symptoms who were identified to have bladder lesions and underwent endoscopic ablation. A Hunner's lesion was identified as an area of erythema that reproduced the patients' pain when touched by the cystoscope. Pathology reports were reviewed and improvement in pain was used as the main outcome measure. RESULTS: Of the 14 patients, 12 had more than 50% symptomatic improvement and 8 patients reported 100% improvement. Mean improvement was 76%. In all patients who improved, the biopsy specimen showed inflammatory cystitis, often with epithelial denudation. Four patients had symptomatic recurrence, but all had improvement after repeat ablation. CONCLUSION: Endoscopic ablation of Hunner's lesions improves symptoms in IC patients. Recurrence of symptoms should prompt repeat cystoscopy to identify recurrent lesions, as repeat ablation offers symptomatic improvement. PMID- 20019979 TI - Arguments against investing widely in robotic prostatectomy in Canada: a wrong focus on tool box rather than surgical expertise. PMID- 20019978 TI - Steps in the investigation and management of low semen volume in the infertile man. AB - An adequate semen volume of ejaculate fluid is required to transport sperm into the female reproductive tract and allow for fertilization of the oocyte. Thus, seminal fluid volume is an important part of the semen analysis done to investigate male infertility. In this article, we review the anatomy and physiology of ejaculation, the various etiologies of low-volume ejaculation (artifactual, structural, functional). We then present a comprehensive algorithm for the evaluation, diagnosis and treatment of the infertile man presenting with low semen volume. PMID- 20019980 TI - Robotic prostatectomy: The new standard of care or a marketing success? PMID- 20019981 TI - Screening for bladder cancer: the best opportunity to reduce mortality. AB - Bladder cancer kills more women than cervical cancer and is also a significant cause of mortality in men. Little progress has been made in improving survival in patients with advanced bladder cancer. Two pilot studies using microhematuria screening have shown that screening for bladder cancer results in close to 80% downstaging, with high-grade cancers being detected before they have invaded the bladder wall. Results of long-term follow-up even suggest a striking reduction in bladder cancer mortality. The main obstacles to screening for bladder cancer may be overcome if a higher-risk population is identified by designing a risk scale for exposure to cigarette smoke and occupational carcinogens, and through genetic testing for susceptibility to cancer and home hematuria screening, which in itself identifies a population with approximately 3% to 4% risk of bladder cancer. The feasibility and cost effectiveness of screening for bladder cancer can be significantly improved by incorporating a secondary screening strategy using a more sensitive and specific bladder cancer marker that is currently available, and by limiting urological evaluations to patients who show positive results on one or more of these tests. Bladder cancer is the most costly cancer to treat in the United States and pharmacoeconomic studies suggest that screening for bladder cancer could not only save lives but also reduce costs per year-life saved. A pilot study is underway and the urology community should be very supportive of studies to validate this opportunity. PMID- 20019982 TI - Chemoprevention in bladder cancer: What's new? AB - Bladder cancer is the sixth most common tumour in Canada and ranks eighth in terms of cancer mortality. Up to now, management of this condition relied mostly on surgical and intravesical treatments once the disease is established. Chemoprevention is an attractive option to prevent the disease in high-risk populations and may well reduce the costs related to its treatment. This review examines the available data on chemoprevention strategies in bladder cancer, with special emphasis on randomized controlled trials when available. PMID- 20019983 TI - The optimal management of T1 high-grade bladder cancer. AB - Stage T1Hg bladder cancer should be considered an aggressive and potentially lethal disease. The importance of initial re-resection to identify unrecognized muscle-invasive disease is significant. Most patients with high-risk disease are candidates for initial bladder salvage with intravesical bacillus Calmette-Guerin vaccine for immunotherapy, a procedure with a high survival rate; however, failure of the procedure may result in a guarded prognosis. Even after apparent success, patients should be informed of the risks of the disease progressing to muscle-invasive or metastatic disease and the need for vigilant monitoring. Despite optimal management, a significant number of patients relapse or progress to invasive disease requiring cystectomy. This review provides insight into the optimal management of T1 high-grade bladder cancer. PMID- 20019984 TI - Histologic variants of urothelial bladder cancer and nonurothelial histology in bladder cancer. AB - Bladder cancer can be classified histologically as urothelial or non-urothelial. Urothelial cancer has a propensity for divergent differentiation, which has increasingly been recognized in recent years due to heightened awareness and improved immunohistochemistry techniques. Furthermore, the recent World Health Organization classification of urothelial cancers improved clarity on this issue, with its listing of 13 histologic variants of urothelial cancer. The divergent differentiation patterns include, amongst others, squamous, glandular, micropapillary, nested, lymphepithelioma-like, plasmacytoid and sarcomatoid variants of urothelial cancer. Attempts to quantify the amount of divergent differentiation present, such as using the nonconventional differentiation number, have been made recently, which will improve the ability to compare publications from different centres. Genetic-based studies have indicated that the histologic variants of urothelial cancer arise from a common clonal precursor. Mostly, the current evidence suggests that urothelial cancer with divergent differentiation has a worse prognosis when compared with pure urothelial cancer. This article will review the current literature on variant histologies of urothelial cancer, and well as new developments in pure squamous cell carcinoma, small cell carcinoma and adenocarcinoma of the bladder. PMID- 20019986 TI - Role of lymphadenectomy for invasive bladder cancer. AB - Radical cystectomy with lymph node dissection remains the standard of care in the treatment of muscle-invasive and refractory non-invasive bladder cancer. Over the past decade, the extent of lymphadenectomy has varied to include dissection up to the common iliac vessels and aortic bifurcation proximally (may also extend up to the level of the inferior mesenteric artery), the genitofemoral nerve laterally, the circumflex iliac vein and lymph node of Cloquet distally, and the hypogastric vessels posteriorly (obturator fossa, presciatic nodes bilaterally and the presacral lymph nodes over the sacral promontory). Evidence supports the role of lymphadenectomy as both a therapeutic and prognostic variable in patients with invasive bladder cancer. We review the literature regarding the role and extent of lymphadenectomy, as well as its impact on patient outcomes. PMID- 20019985 TI - The management of BCG failure in non-muscle-invasive bladder cancer: an update. AB - Up to 40% of patients with non-muscle-invasive bladder cancer (NMIBC) will fail intravesical bacillus Calmette-Guerin (BCG) therapy. There is unfortunately no current gold standard for salvage intravesical therapy after appropriate BCG treatment. Indeed, outcomes are at best suboptimal. The vast majority of low grade NMIBC are prone to recur but very rarely progress. Failure after intravesical BCG in these patients is usually superficial and low-grade. At the other end of the spectrum, failure to respond to BCG in high-risk T1 bladder cancer and/or carcinoma in situ (CIS or TIS) is more problematic, since those tumours often have the potential to progress to muscle invasion. In these cases, radical cystectomy remains the mainstay after BCG failure. With appropriate selection, certain patients who "fail" BCG (but with favourable risk factors) can be managed with intravesical regimens, including repeated BCG, BCG plus cytokines, intravesical chemotherapy, thermochemotherapy or new immunotherapeutic modalities. In this review, reasons explaining BCG failure, how to define BCG failure, optimal risk stratification and prediction of response and management of BCG failures are discussed. PMID- 20019987 TI - Management of the urethra in urothelial bladder cancer. AB - The standard of care in the management of invasive urothelial cancer of the bladder is radical cystectomy and pelvic lymphadenectomy. Although uncommon, recurrence of disease in the retained urethra following cystectomy carries a poor prognosis. The need for assessment of risk of recurrence is greater now than ever with wider adoption of orthotopic bladder substitution. This review will address the contemporary management of the urethra following cystectomy for urothelial cancer. PMID- 20019988 TI - The case for prostate capsule-sparing radical cystectomy in selected patients. PMID- 20019989 TI - Is sparing the prostate still considered radical cystectomy? The case against prostate-sparing cystectomy for bladder cancer. PMID- 20019990 TI - Perioperative chemotherapy for muscle-invasive bladder cancer. AB - Considerable debate exists concerning the combined use of systemic chemotherapy and radical surgery for muscle-invasive bladder cancer. While there is evidence for a survival benefit after neoadjuvant chemotherapy, the benefit is modest and the potential toxicity and delay of time to surgery prior to cystectomy appears to be deterring many surgeons from its administration. The evidence for adjuvant chemotherapy, on the other hand, is less compelling and substantial. Furthermore, the role of adjuvant compared to salvage chemotherapy requires further investigation. Similarly, research continues on identifying molecular and clinical markers to best stratify patients for optimal perioperative therapy. In this article, the evidence for radical cystectomy and chemotherapy, given either in a neoadjuvant or adjuvant setting, will be reviewed. PMID- 20019991 TI - Management of metastatic urothelial cancer: the role of surgery as an adjunct to chemotherapy. AB - Metastatic or unresectable disease is identified in approximately 20% of patients presenting with invasive urothelial cancer. In addition, up to 50% of patients will develop metastases following radical cystectomy for clinically localized disease. Multiagent cisplatin-based chemotherapy is considered standard first line treatment for these patients. Although urothelial cancer is considered a chemosensitive tumour, metastatic disease is associated with poor prognosis and short-term survival. Here, we review the role of a multidisciplinary approach to treating patients with metastatic urothelial cancer. PMID- 20019992 TI - Personalized medicine in advanced urothelial cancer: when to treat, how to treat and who to treat. AB - The past decade has provided an improved understanding of the molecular mechanism of bladder cancer by defining distinct pathways in tumorigenesis and progression. Advances in technologies, such as high-throughput transcript profiling, microarrays and proteomics, offer a systematic approach to identifying targets for bladder cancer diagnostics and drug discovery. This review presents a select outline of the advances in the development of bio-markers and targets for patient prognosis and therapy selection. This paper describes a representative cohort of recent studies that have the potential to significantly impact the management of muscle invasive and metastatic urothelial carcinoma of the bladder. Space constraints do not permit this review to be comprehensive and we apologize to the authors whose work we do not cite. PMID- 20019994 TI - Suffer the loss, but remember the man. PMID- 20019993 TI - Surveillance strategies after definitive therapy of invasive bladder cancer. AB - Following definitive therapy for muscle invasive bladder cancer, patients remain at risk for local and distant recurrence. Additionally, recurrences can result from formation of new tumours elsewhere in the urinary tract. We review patterns of recurrence and the prognosis associated with recurrence. Optimal surveillance strategies are discussed. PMID- 20019995 TI - Better drug therapy for children: Time for action. PMID- 20019996 TI - The physician's role and responsibility in reporting adverse drug events in children. PMID- 20019998 TI - How to improve your diagnostic tools for genetic disorders. PMID- 20019997 TI - Ethics and the publication of research. PMID- 20019999 TI - Paediatric adverse drug reactions can be fatal. PMID- 20020000 TI - Where physicians look for information on drug prescribing for children. AB - BACKGROUND: Despite the important role of drug therapy in children, there is often a lack of readily available information regarding the indications and dosing regimens for medications in paediatrics. OBJECTIVE: To collect data on where family physicians obtain this prescribing information. METHOD: A structured questionnaire was mailed to 500 family physicians in Ontario. RESULTS: Questionnaires were returned by 261 (52%) family physicians, 217 (83%) of whom identified themselves as currently involved in the care of children. Most (87%) reported that the Compendium of Pharmaceuticals and Specialties (CPS) was the source that they most commonly consulted for drug information in children. The available sources of information on prescribing for children were thought to be not adequate by 40% and not readily available by 27%. Sixty-one per cent reported being moderately confident (in doubt part of the time) about their decisions regarding drug prescribing in this age group. The majority (70%) had learned most of what they know about prescribing in paediatrics during practice, while 69% and 62% reported they had little or no teaching during undergraduate and postgraduate (internship or residency) medical education, respectively. CONCLUSIONS: Although it is recognized that for a number of drugs used in children the CPS does not reflect the current standard of care in paediatrics, it is currently the source most commonly referred to by family physicians. Further work should be done in the provision of useful information on paediatric drug therapy to family physicians. PMID- 20020001 TI - Does topical amethocaine gel reduce pain from heel prick blood sampling in premature infants? A randomized double-blind cross-over controlled study. AB - BACKGROUND: Heel prick blood sampling is the most common painful invasive procedure performed on neonates. Currently, there are no effective ways to provide pain relief from this painful procedure. OBJECTIVE: To assess the efficacy of the topical anesthetic amethocaine 4% gel (Ametop, Smith & Nephew Inc, St Laurent) in reducing the pain of heel prick blood sampling in neonates. METHODS: A randomized, double-blind, placebo controlled, crossover trial was conducted. Neonates between 33 to 37 weeks' gestational age in their first seven days of life were eligible. Heel prick blood sampling was performed on each participant twice. Each infant was randomly assigned to receive either amethocaine 4% gel or placebo to the heel for the first prick, and then received the alternative agent for the second prick. Prick pain was assessed using both Premature Infant Pain Profile (PIPP) and Neonatal Infant Pain Scale (NIPS). Squeeze pain was assessed by NIPS. RESULTS: Ten babies were recruited. There were no significant differences in the average PIPP and NIPS scores between the treatment and placebo groups for both prick and squeeze pains from heel prick blood sampling. For prick pain, linear-regression showed significant correlation between the PIPP and NIPS scores. No adverse reactions were observed after application of either the active or placebo agents. CONCLUSION: Topical amethocaine 4% gel is not shown to reduce prick and squeeze pains significantly from heel prick blood sampling in neonates between 33 to 37 weeks' gestational age. Further studies are needed to find ways to provide effective pain relief from this common procedure. PMID- 20020002 TI - The child with multiple short courses of steroid therapy. PMID- 20020003 TI - Are home visiting programs more effective than the standard of care at preventing injury in children who are at risk for injury?: Part A. PMID- 20020004 TI - Drug investigation for Canadian children: The role of the Canadian Paediatric Society. PMID- 20020005 TI - Alternative treatments for attention deficit hyperactivity disorder. PMID- 20020006 TI - Childhood tuberculosis: In Canada and around the world. PMID- 20020007 TI - Transfusion and risk of infection in Canada. PMID- 20020009 TI - Tuberculosis in Canada: Global view and new challenges. PMID- 20020010 TI - "To BCG or not to BCG, that is the question!". The challenge of BCG vaccination: Why can't we get it right? PMID- 20020011 TI - Clinical predictors of positive urine cultures in young children at risk for urinary tract infection. AB - BACKGROUND: Urinary tract infections (UTIs) are a common source of bacterial infection among young febrile children. The diagnosis of UTI is challenging because the clinical presentation is not specific. OBJECTIVE: To describe clinical predictors to identify young children needing urine culture for evaluation of UTI. METHODS: Retrospective cohort study of all children younger than two years of age (719 hospital visits for 545 patients) suspected of having a UTI during a 12-month period. The outcome was UTI, defined as a catheterized urine culture with pure growth of 104 colonies/mL or greater, or suprapubic aspiration culture with 103 colonies/mL or greater. Candidate predictors included demographic, historical and physical examination variables. RESULTS: The medical records of 545 children younger than two years of age were reviewed. Forty-six per cent were girls. Mean age was 9.1 months (SD 7 months). Four variables were found to predict UTI: absence of another source of fever on examination (odds ratio [OR]=41.6 [95% CI, 8.8 to 197.4]), foul smelling urine (OR=19.7 [95% CI, 5.7 to 68.2]), white blood cell count greater than 15,000/mm3 (OR=4.3 [95% CI, 2.0 to 9.3]), younger than six months old (OR=3.1 [95% CI, 1.3 to 7.1]). The sensitivity of an abnormal urine analysis was 0.77 (95% CI, 0.66 to 0.88) and the specificity was 0.31 (95% CI, 0.2 to 0.42). CONCLUSION: An incremental increase in risk for UTI is associated with younger age (younger than six months), having a white blood cell count higher than 15,000/mm3, parental report of malodorous or foul smelling urine and the absence of an alternative source of fever. In the present patient population, obtaining a urine culture from children with at least one of these clinical predictors would have resulted in missing one UTI (2%), and 111 negative cultures (20%) would have been avoided. PMID- 20020012 TI - CPSP Highlights. PMID- 20020013 TI - [Not Available]. AB - Congenital insensitivity to pain is a rare illness that can cause accidental external and internal injuries in children. When child abuse is suspected, the awareness of this illness can complicate any medicolegal interpretation of findings.Several components are needed to diagnose congenital insensitivity to pain: lack of sensitivity to pain since birth; involvement of the whole body; integrity of osteotendinous reflexes; and other sensitivity processes and negative paraclinical, including histological, examinations.This article describes the case of two siblings examined during an emergency legal procedure at the forensic medicine unit of the Grenobles University Hospital Centre in France. The medico-legal interpretation of facts and examinations must be conservative in case of legal complications and must follow an analysis of the child's and family's medicosocial problems since the child's birth. The family's socioeducational support and the communication of information on this illness should be a priority to avoid any misunderstanding and overrun in the child's management and monitoring. PMID- 20020014 TI - Paediatric chairs of Canada: academic paediatric workforce survey - report for the 1999/2000 academic year. PMID- 20020016 TI - Clinical scenarios in paediatric tuberculosis - case presentations. PMID- 20020015 TI - A clinical approach to paediatric tuberculosis in Canada. AB - OBJECTIVE: To review clinical aspects of management of tuberculosis (TB) infection and disease in Canadian children in the context of the global TB epidemic and the rising incidence of drug-resistant TB. DATA SOURCES: ORIGINAL AND REVIEW ARTICLES PERTINENT TO: epidemiology of TB globally and in Canada; management of latent TB infection and TB disease in children; diagnostic tests for latent TB infection and TB disease; and management of drug-resistant TB disease. Multiple Medline searches were used including combinations of the MeSH terms 'Tuberculosis*' (and its multiple subheadings), 'Child*', 'Drug Resistance', 'Mycobacterium tuberculosis*' and 'Canada/epidemiology*'. Select relevant textbooks were reviewed. DATA SELECTION AND EXTRACTION: The articles were analyzed from the perspective of clinicians managing children in Canada today, and from our experience of managing children with TB in Southern Ontario. DATA SYNTHESIS: TB in Canada is largely a disease of the foreign-born and their children, but continues to occur in aboriginal children. Drug resistance is increasing globally and in Canada. Most children with TB disease in Canada are asymptomatic and found through contact tracing. False positive skin tests are frequent where TB prevalence is low. CONCLUSIONS: Obtain source case drug sensitivities when treating TB contacts and those with latent TB infection. Obtain cultures before treating TB disease and treat disease with at least four antituberculous drugs while awaiting sensitivities. Use Directly Observed Therapy for TB disease. Confine TB skin testing to children at high risk for TB infection or disease, including contacts of infectious patients and recent immigrants. A team approach and infection control measures including environmental controls are important in managing TB disease. PMID- 20020017 TI - Continuing medical education - quiz. PMID- 20020018 TI - Clinician's Corner. PMID- 20020019 TI - Emergency contraception. PMID- 20020020 TI - Emergency contraception: Preventing pregnancy after you have had sex. PMID- 20020021 TI - The science of environmental influence. PMID- 20020022 TI - Putting media under the microscope: Understanding and challenging media's influence on the health and well-being of children and youth. PMID- 20020023 TI - Enuresis: A maturational lag. PMID- 20020024 TI - West Nile virus - Mosquitoes no longer just an annoyance! PMID- 20020025 TI - West Nile virus: Information for parents. PMID- 20020027 TI - What physicians and parents should know about the Internet. PMID- 20020026 TI - The digital divide - a new generation gap. Parental knowledge of their children's Internet use. PMID- 20020028 TI - Functional brain imaging: Evaluation of the effects of violent media exposure. PMID- 20020029 TI - Teens, sex and the media: Is there a connection? PMID- 20020030 TI - The impact of the media on eating disorders in children and adolescents. PMID- 20020031 TI - [Not Available]. AB - Over the past twenty years, the question of the effects of violence on television has figured prominently in public opinion and hundreds of studies have been devoted to this subject. Many researchers have determined that violence has a negative impact on behaviour. The public, broadcasters and political figures all support the idea of reducing the total amount of violence on television - in particular in shows for children. A thousand programs aired between 1993 and 2001 on major non-specialty television networks in Canada were analyzed: TVA, TQS, as well as CTV and Global, private French and English networks, as well as the English CBC Radio and French Radio-Canada for the public networks. The methodology consists of a classic analysis of content where an act of violence constitutes a unit of analysis. The data collected revealed that the amount of violence has increased regularly since 1993 despite the stated willingness on the part of broadcasters to produce programs with less violence. The total number of violent acts, as well as the number of violent acts per hour, is increasing. Private networks deliver three times more violence than public networks. Researchers have also noted that a high proportion of violence occurs in programs airing before 2100 hours, thereby exposing a large number of children to this violence. Finally, the author would also like to indicate that psychological violence is taking on a more significant role. PMID- 20020032 TI - Le casque de velo. PMID- 20020033 TI - Call for new studies: Research opportunities. PMID- 20020034 TI - Impact of media use on children and youth. PMID- 20020035 TI - Managing media use in the home. PMID- 20020036 TI - Hypothalamic Sirt1 regulates food intake in a rodent model system. AB - Sirt1 is an evolutionarily conserved NAD(+) dependent deacetylase involved in a wide range of processes including cellular differentiation, apoptosis, as well as metabolism, and aging. In this study, we investigated the role of hypothalamic Sirt1 in energy balance. Pharmacological inhibition or siRNA mediated knock down of hypothalamic Sirt1 showed to decrease food intake and body weight gain. Central administration of a specific melanocortin antagonist, SHU9119, reversed the anorectic effect of hypothalamic Sirt1 inhibition, suggesting that Sirt1 regulates food intake through the central melanocortin signaling. We also showed that fasting increases hypothalamic Sirt1 expression and decreases FoxO1 (Forkhead transcription factor) acetylation suggesting that Sirt1 regulates the central melanocortin system in a FoxO1 dependent manner. In addition, hypothalamic Sirt1 showed to regulate S6K signaling such that inhibition of the fasting induced Sirt1 activity results in up-regulation of the S6K pathway. Thus, this is the first study providing a novel role for the hypothalamic Sirt1 in the regulation of food intake and body weight. Given the role of Sirt1 in several peripheral tissues and hypothalamus, potential therapies centered on Sirt1 regulation might provide promising therapies in the treatment of metabolic diseases including obesity. PMID- 20020037 TI - A comparison of the wellbeing of orphans and abandoned children ages 6-12 in institutional and community-based care settings in 5 less wealthy nations. AB - BACKGROUND: Leaders are struggling to care for the estimated 143,000,000 orphans and millions more abandoned children worldwide. Global policy makers are advocating that institution-living orphans and abandoned children (OAC) be moved as quickly as possible to a residential family setting and that institutional care be used as a last resort. This analysis tests the hypothesis that institutional care for OAC aged 6-12 is associated with worse health and wellbeing than community residential care using conservative two-tail tests. METHODOLOGY: The Positive Outcomes for Orphans (POFO) study employed two-stage random sampling survey methodology in 6 sites across 5 countries to identify 1,357 institution-living and 1,480 community-living OAC ages 6-12, 658 of whom were double-orphans or abandoned by both biological parents. Survey analytic techniques were used to compare cognitive functioning, emotion, behavior, physical health, and growth. Linear mixed-effects models were used to estimate the proportion of variability in child outcomes attributable to the study site, care setting, and child levels and institutional versus community care settings. Conservative analyses limited the community living children to double-orphans or abandoned children. PRINCIPAL FINDINGS: Health, emotional and cognitive functioning, and physical growth were no worse for institution-living than community-living OAC, and generally better than for community-living OAC cared for by persons other than a biological parent. Differences between study sites explained 2-23% of the total variability in child outcomes, while differences between care settings within sites explained 8-21%. Differences among children within care settings explained 64-87%. After adjusting for sites, age, and gender, institution vs. community-living explained only 0.3-7% of the variability in child outcomes. CONCLUSION: This study does not support the hypothesis that institutional care is systematically associated with poorer wellbeing than community care for OAC aged 6-12 in those countries facing the greatest OAC burden. Much greater variability among children within care settings was observed than among care settings type. Methodologically rigorous studies must be conducted in those countries facing the new OAC epidemic in order to understand which characteristics of care promote child wellbeing. Such characteristics may transcend the structural definitions of institutions or family homes. PMID- 20020038 TI - Nanoscale imaging of whole cells using a liquid enclosure and a scanning transmission electron microscope. AB - Nanoscale imaging techniques are needed to investigate cellular function at the level of individual proteins and to study the interaction of nanomaterials with biological systems. We imaged whole fixed cells in liquid state with a scanning transmission electron microscope (STEM) using a micrometer-sized liquid enclosure with electron transparent windows providing a wet specimen environment. Wet-STEM images were obtained of fixed E. coli bacteria labeled with gold nanoparticles attached to surface membrane proteins. Mammalian cells (COS7) were incubated with gold-tagged epidermal growth factor and fixed. STEM imaging of these cells resulted in a resolution of 3 nm for the gold nanoparticles. The wet-STEM method has several advantages over conventional imaging techniques. Most important is the capability to image whole fixed cells in a wet environment with nanometer resolution, which can be used, e.g., to map individual protein distributions in/on whole cells. The sample preparation is compatible with that used for fluorescent microscopy on fixed cells for experiments involving nanoparticles. Thirdly, the system is rather simple and involves only minimal new equipment in an electron microscopy (EM) laboratory. PMID- 20020039 TI - An information gap in DNA evidence interpretation. AB - Forensic DNA evidence often contains mixtures of multiple contributors, or is present in low template amounts. The resulting data signals may appear to be relatively uninformative when interpreted using qualitative inclusion-based methods. However, these same data can yield greater identification information when interpreted by computer using quantitative data-modeling methods. This study applies both qualitative and quantitative interpretation methods to a well characterized DNA mixture and dilution data set, and compares the inferred match information. The results show that qualitative interpretation loses identification power at low culprit DNA quantities (below 100 pg), but that quantitative methods produce useful information down into the 10 pg range. Thus there is a ten-fold information gap that separates the qualitative and quantitative DNA mixture interpretation approaches. With low quantities of culprit DNA (10 pg to 100 pg), computer-based quantitative interpretation provides greater match sensitivity. PMID- 20020040 TI - Next-generation phage display: integrating and comparing available molecular tools to enable cost-effective high-throughput analysis. AB - BACKGROUND: Combinatorial phage display has been used in the last 20 years in the identification of protein-ligands and protein-protein interactions, uncovering relevant molecular recognition events. Rate-limiting steps of combinatorial phage display library selection are (i) the counting of transducing units and (ii) the sequencing of the encoded displayed ligands. Here, we adapted emerging genomic technologies to minimize such challenges. METHODOLOGY/PRINCIPAL FINDINGS: We gained efficiency by applying in tandem real-time PCR for rapid quantification to enable bacteria-free phage display library screening, and added phage DNA next generation sequencing for large-scale ligand analysis, reporting a fully integrated set of high-throughput quantitative and analytical tools. The approach is far less labor-intensive and allows rigorous quantification; for medical applications, including selections in patients, it also represents an advance for quantitative distribution analysis and ligand identification of hundreds of thousands of targeted particles from patient-derived biopsy or autopsy in a longer timeframe post library administration. Additional advantages over current methods include increased sensitivity, less variability, enhanced linearity, scalability, and accuracy at much lower cost. Sequences obtained by qPhage plus pyrosequencing were similar to a dataset produced from conventional Sanger sequenced transducing-units (TU), with no biases due to GC content, codon usage, and amino acid or peptide frequency. These tools allow phage display selection and ligand analysis at >1,000-fold faster rate, and reduce costs approximately 250-fold for generating 10(6) ligand sequences. CONCLUSIONS/SIGNIFICANCE: Our analyses demonstrates that whereas this approach correlates with the traditional colony-counting, it is also capable of a much larger sampling, allowing a faster, less expensive, more accurate and consistent analysis of phage enrichment. Overall, qPhage plus pyrosequencing is superior to TU-counting plus Sanger sequencing and is proposed as the method of choice over a broad range of phage display applications in vitro, in cells, and in vivo. PMID- 20020041 TI - Identifying blood biomarkers and physiological processes that distinguish humans with superior performance under psychological stress. AB - BACKGROUND: Attrition of students from aviation training is a serious financial and operational concern for the U.S. Navy. Each late stage navy aviator training failure costs the taxpayer over $1,000,000 and ultimately results in decreased operational readiness of the fleet. Currently, potential aviators are selected based on the Aviation Selection Test Battery (ASTB), which is a series of multiple-choice tests that evaluate basic and aviation-related knowledge and ability. However, the ASTB does not evaluate a person's response to stress. This is important because operating sophisticated aircraft demands exceptional performance and causes high psychological stress. Some people are more resistant to this type of stress, and consequently better able to cope with the demands of naval aviation, than others. METHODOLOGY/PRINCIPAL FINDINGS: Although many psychological studies have examined psychological stress resistance none have taken advantage of the human genome sequence. Here we use high-throughput -omic biology methods and a novel statistical data normalization method to identify plasma proteins associated with human performance under psychological stress. We identified proteins involved in four basic physiological processes: innate immunity, cardiac function, coagulation and plasma lipid physiology. CONCLUSIONS/SIGNIFICANCE: The proteins identified here further elucidate the physiological response to psychological stress and suggest a hypothesis that stress-susceptible pilots may be more prone to shock. This work also provides potential biomarkers for screening humans for capability of superior performance under stress. PMID- 20020042 TI - Clustering of socioeconomic, behavioural, and neonatal risk factors for infant health in pregnant smokers. AB - BACKGROUND: Tobacco smoking is a major cause of morbidity and mortality, including during pregnancy. Although effective ways of promoting smoking cessation during pregnancy exist, the impact of these interventions has not been studied at a national level. We estimated the prevalence of smoking throughout pregnancy in the Netherlands and quantified associations of maternal smoking throughout pregnancy with socioeconomic, behavioural, and neonatal risk factors for infant health and development. METHODOLOGY/PRINCIPAL FINDINGS: Data of five national surveys, containing records of 14,553 Dutch mothers and their offspring were analyzed. From 2001 to 2007, the overall rate of smoking throughout pregnancy fell by 42% (from 13.2% to 7.6%) mainly as a result of a decrease among highly educated women. In the lowest-educated group, the overall rate of smoking throughout pregnancy was six times as high as in the highest-educated group (18.7% versus 3.2%). Prenatal tobacco smoke exposure was associated with increased risk of extremely preterm (17,000 cDNA clones, and >2,000 kinase- targeted siRNAs against a 5.6 kb luciferase minigene construct, encompassing exon 71 to exon 73 of human dystrophin. As a result, we identified several enhancers of exon skipping, acting on both the reporter construct as well as endogenous dystrophin in mdx cells. Multiple mechanisms of action were identified, including histone deacetylase inhibition, tubulin modulation and pre-mRNA processing. Among others, the nucleolar protein NOL8 and staufen RNA binding protein homolog 2 (Stau2) were found to induce endogenous exon skipping in mdx cells in an AON-dependent fashion. An unexpected but recurrent theme observed in our screening efforts was the apparent link between the inhibition of cell cycle progression and the induction of exon skipping. PMID- 20020057 TI - Aurintricarboxylic acid is a potent inhibitor of influenza A and B virus neuraminidases. AB - BACKGROUND: Influenza viruses cause serious infections that can be prevented or treated using vaccines or antiviral agents, respectively. While vaccines are effective, they have a number of limitations, and influenza strains resistant to currently available anti-influenza drugs are increasingly isolated. This necessitates the exploration of novel anti-influenza therapies. METHODOLOGY/PRINCIPAL FINDINGS: We investigated the potential of aurintricarboxylic acid (ATA), a potent inhibitor of nucleic acid processing enzymes, to protect Madin-Darby canine kidney cells from influenza infection. We found, by neutral red assay, that ATA was protective, and by RT-PCR and ELISA, respectively, confirmed that ATA reduced viral replication and release. Furthermore, while pre-treating cells with ATA failed to inhibit viral replication, pre-incubation of virus with ATA effectively reduced viral titers, suggesting that ATA may elicit its inhibitory effects by directly interacting with the virus. Electron microscopy revealed that ATA induced viral aggregation at the cell surface, prompting us to determine if ATA could inhibit neuraminidase. ATA was found to compromise the activities of virus-derived and recombinant neuraminidase. Moreover, an oseltamivir-resistant H1N1 strain with H274Y was also found to be sensitive to ATA. Finally, we observed additive protective value when infected cells were simultaneously treated with ATA and amantadine hydrochloride, an anti-influenza drug that inhibits M2-ion channels of influenza A virus. CONCLUSIONS/SIGNIFICANCE: Collectively, these data suggest that ATA is a potent anti-influenza agent by directly inhibiting the neuraminidase and could be a more effective antiviral compound when used in combination with amantadine hydrochloride. PMID- 20020058 TI - P21cip-overexpression in the mouse beta cells leads to the improved recovery from streptozotocin-induced diabetes. AB - Under normal conditions, the regeneration of mouse beta cells is mainly dependent on their own duplication. Although there is evidence that pancreatic progenitor cells exist around duct, whether non-beta cells in the islet could also potentially contribute to beta cell regeneration in vivo is still controversial. Here, we developed a novel transgenic mouse model to study the pancreatic beta cell regeneration, which could specifically inhibit beta cell proliferation by overexpressing p21(cip) in beta cells via regulation of the Tet-on system. We discovered that p21 overexpression could inhibit beta-cell duplication in the transgenic mice and these mice would gradually suffer from hyperglycemia. Importantly, the recovery efficiency of the p21-overexpressing mice from streptozotocin-induced diabetes was significantly higher than control mice, which is embodied by better physiological quality and earlier emergence of insulin expressing cells. Furthermore, in the islets of these streptozotocin-treated transgenic mice, we found a large population of proliferating cells which expressed pancreatic duodenal homeobox 1 (PDX1) but not markers of terminally differentiated cells. Transcription factors characteristic of early pancreatic development, such as Nkx2.2 and NeuroD1, and pancreatic progenitor markers, such as Ngn3 and c-Met, could also be detected in these islets. Thus, our work showed for the first time that when beta cell self-duplication is repressed by p21 overexpression, the markers for embryonic pancreatic progenitor cells could be detected in islets, which might contribute to the recovery of these transgenic mice from streptozotocin-induced diabetes. These discoveries could be important for exploring new diabetes therapies that directly promote the regeneration of pancreatic progenitors to differentiate into islet beta cells in vivo. PMID- 20020059 TI - Retraction: an African ancestry-specific allele of CTLA4 confers protection against rheumatoid arthritis in African Americans. PMID- 20020060 TI - Binding of NIR-conPK and NIR-6T to astrocytomas and microglial cells: evidence for a protein related to TSPO. AB - PK 11195 and DAA1106 bind with high-affinity to the translocator protein (TSPO, formerly known as the peripheral benzodiazepine receptor). TSPO expression in glial cells increases in response to cytokines and pathological stimuli. Accordingly, [(11)C]-PK 11195 and [(11)C]-DAA1106 are recognized molecular imaging (MI) agents capable of monitoring changes in TSPO expression occurring in vivo and in response to various neuropathologies.Here we tested the pharmacological characteristics and TSPO-monitoring potential of two novel MI agents: NIR-conPK and NIR-6T. NIR-conPK is an analogue of PK 11195 conjugated to the near-infrared (NIR) emitting fluorophore: IRDye 800CW. NIR-6T is a DAA1106 analogue also conjugated to IRDye 800CW.We found that NIR-6T competed for [(3)H] PK 11195 binding in astrocytoma cell homogenates with nanomolar affinity, but did not exhibit specific binding in intact astrocytoma cells in culture, indicating that NIR-6T is unlikely to constitute a useful MI agent for monitoring TSPO expression in intact cells. Conversely, we found that NIR-conPK did not compete for [(3)H]-PK 11195 binding in astrocytoma cell homogenate, but exhibited specific binding in intact astrocytoma cells in culture with nanomolar affinity, suggesting that NIR-conPK binds to a protein distinct, but related to, TSPO. Accordingly, treating intact astrocytoma cells and microglia in culture with cytokines led to significant changes in the amount of NIR-conPK specific binding without corresponding change in TSPO expression. Remarkably, the cytokine-induced changes in the protein targeted by NIR-conPK in intact microglia were selective, since IFN-gamma (but not TNFalpha and TGFbeta) increased the amount of NIR-conPK specific binding in these cells.Together these results suggest that NIR-conPK binds to a protein that is related to TSPO, and expressed by astrocytomas and microglia. Our results also suggest that the expression of this protein is increased by specific cytokines, and thus allows for the monitoring of a particular subtype of microglia activation. PMID- 20020062 TI - Brownie, a gene involved in building complex respiratory devices in insect eggshells. AB - BACKGROUND: Insect eggshells must combine protection for the yolk and embryo with provisions for respiration and for the entry of sperm, which are ensured by aeropyles and micropyles, respectively. Insects which oviposit the eggs in an egg case have a double problem of respiration as gas exchange then involves two barriers. An example of this situation is found in the cockroach Blattella germanica, where the aeropyle and the micropyle are combined in a complex structure called the sponge-like body. The sponge-like body has been well described morphologically, but nothing is known about how it is built up. METHODOLOGY/PRINCIPAL FINDINGS: In a library designed to find genes expressed during late chorion formation in B. germanica, we isolated the novel sequence Bg30009 (now called Brownie), which was outstanding due to its high copy number. In the present work, we show that Brownie is expressed in the follicle cells localized in the anterior pole of the oocyte in late choriogenesis. RNA interference (RNAi) of Brownie impaired correct formation of the sponge-like body and, as a result, the egg-case was also ill-formed and the eggs were not viable. CONCLUSIONS/SIGNIFICANCE: Results indicate that the novel gene Brownie plays a pivotal role in building up the sponge-like body. Brownie is the first reported gene involved in the construction of complex eggshell respiratory structures. PMID- 20020061 TI - KRAS mutations in primary colorectal cancer tumors and related metastases: a potential role in prediction of lung metastasis. AB - BACKGROUND: KRAS mutations in colorectal cancer primary tumors predict resistance to anti-Epidermal Growth Factor Receptor (EGFR) monoclonal antibody therapy in patients with metastatic colorectal cancer, and thus represent a true indicator of EGFR pathway activation status. METHODOLOGY/PRINCIPAL FINDINGS: KRAS mutations were retrospectively studied using polymerase chain reactions and subsequent sequencing of codons 12 and 13 (exon 2) in 110 patients with metastatic colorectal tumors. These studies were performed using tissue samples from both the primary tumor and their related metastases (93 liver, 84%; 17 lung, 16%). All patients received adjuvant 5-Fluorouracil-based polychemotherapy after resection of metastases. None received anti-EGFR therapy. Mutations in KRAS were observed in 37 (34%) of primary tumors and in 40 (36%) of related metastases, yielding a 94% level of concordance (kappa index 0.86). Patients with primary tumors possessing KRAS mutations had a shorter disease-free survival period after metastasis resection (12.0 vs 18.0 months; P = 0.035) than those who did not. A higher percentage of KRAS mutations was detected in primary tumors of patiens with lung metastases than in patients with liver metastases (59% vs 32%; p = 0.054). To further evaluate this finding we analyzed 120 additional patients with unresectable metastatic colorectal cancer who previously had their primary tumors evaluated for KRAS mutational status for clinical purposes. Separately, the analysis of these 120 patients showed a tendency towards a higher degree of KRAS mutations in primary tumors of patients with lung metastases, although it did not reach statistical significance. Taken together the group of 230 patients showed that KRAS was mutated significantly more often in the primary tumors of patients with lung metastases (57% vs 35%; P = 0.006). CONCLUSIONS/SIGNIFICANCE: Our results suggest a role for KRAS mutations in the propensity of primary colorectal tumors to metastasize to the lung. PMID- 20020063 TI - Vector field embryogeny. AB - We present a novel approach toward evolving artificial embryogenies, which omits the graph representation of gene regulatory networks and directly shapes the dynamics of a system, i.e., its phase space. We show the feasibility of the approach by evolving cellular differentiation, a basic feature of both biological and artificial development. We demonstrate how a spatial hierarchy formulation can be integrated into the framework and investigate the evolution of a hierarchical system. Finally, we show how the framework allows the investigation of allometry, a biological phenomenon, and its role for evolution. We find that direct evolution of allometric change, i.e., the evolutionary adaptation of the speed of system states on transient trajectories in phase space, is advantageous for a cellular differentiation task. PMID- 20020075 TI - Correlation of serum and salivary CA125 levels in patients with breast cancer. AB - AIM: The aim of this study was to examine the relationship between serum and saliva levels of cancer antigen (CA) 125 and compare them among healthy women and patients with treated and untreated breast cancer. METHODS AND MATERIALS: CA125 levels were assayed in serum and unstimulated whole saliva of 25 normal healthy women, 24 patients with untreated breast cancer, and 23 patients with treated breast cancer using enzyme immunoassay (EIA) kits. RESULTS: The mean saliva and serum CA125 levels were significantly higher in untreated cancer women compared to healthy and treated groups. The serum and saliva CA125 levels showed a significant modest positive correlation (r=0.38; p=0.01). CONCLUSION: CA125 level in saliva is higher than in serum with a modest positive correlation between each other. CLINICAL SIGNIFICANCE: Serum and salivary CA125 levels were significantly higher in women with untreated breast cancer than healthy women and women who were treated for breast cancer. PMID- 20020064 TI - Fluorescent duplex allele-specific PCR and amplicon melting for rapid homogeneous mtDNA haplogroup H screening and sensitive mixture detection. AB - BACKGROUND: For large scale studies aiming at a better understanding of mitochondrial DNA (mtDNA), sequence variation in particular mt haplogroups (hgs) and population structure, reliable low-cost high-throughput genotyping assays are needed. Furthermore, methods facilitating sensitive mixture detection and relative quantification of allele proportions are indispensable for the study of heteroplasmy, mitochondrial sequence evolution, and mitochondrial disorders. Here the properties of a homogeneous competitive duplex allele specific PCR (ARMS) assay were scrutinized in the light of these requirements. METHODOLOGY/PRINCIPAL FINDINGS: A duplex ARMS assay amplifying either the ancestral mtDNA 2706G allele (non-hg H samples) or the derived 7028C allele (hg H samples) in the presence of SYBR Green fluorescent reporter dye was developed and characterized. Product detection, allele calling, and hg inference were based on the amplicon characteristic melting-point temperatures obtained with on-line post-PCR fluorescent dissociation curve analysis (DCA). The analytical window of the assay covered at least 5 orders of magnitude of template DNA input with a detection limit in the low picogram range of genomic DNA. A set of forensically relevant test specimens was analyzed successfully. The presence of mtDNA mixtures was detected over a broad range of input DNA amounts and mixture ratios, and the estimation of allele proportions in samples with known total mtDNA content was feasible with limitations. A qualified DNA analyst successfully analyzed approximately 2,200 DNA extracts within three regular working days, without using robotic lab-equipment. By performing the amplification on-line, the assay also facilitated absolute mtDNA quantification. CONCLUSIONS: Although this assay was developed just for a particular purpose, the approach is general in that it is potentially suitable in a broad variety of assay-layouts for many other applications, including the analysis of mixtures. Homogeneous ARMS-DCA is a valuable tool for large-volume studies targeting small numbers of single nucleotide polymorphisms (SNPs). PMID- 20020076 TI - Effect of postoperative bleaching on marginal leakage of resin composite and resin-modified glass ionomer restorations at different delayed periods of exposure to carbamide peroxide. AB - AIM: To evaluate the effect of dental bleaching with carbamide peroxide at different exposure times on the microleakage of resin composite and resin modified glass ionomer restorations after placement in extracted human teeth. METHODS AND MATERIALS: 120 Class V cavity preparations were placed at the cementoenamel junction (CEJ) of human teeth. Half of the cavities were restored with Filtek P60 resin composite (C) and the other half were restored with Fuji II LC resin-modified glass ionomer (G). Each group was randomly divided into four subgroups (n=15). Groups C1 and G1 were not bleached and stored in artificial saliva at 37 degrees C to serve as control groups, while in Groups C2 and G2, C3 and G3, and C4 and G4 specimens were exposed to a 15% carbamide peroxide gel for one day, one week, and two weeks, respectively, following the placement of restorations. Microleakage was assessed using the dye penetration method. Data were analyzed using the Kruskal-Wallis and Wilcoxon tests (p=0.05). RESULTS: The Kruskal-Wallis test showed no significant difference among all groups of composite or glass ionomer restorations with either enamel or dentinal margins with regard to microleakage (p>0.05). The Wilcoxon test revealed more marginal leakage in the enamel/glass ionomer margins than the enamel/composite margins (p<0.05). In comparisons within each group, the Wilcoxon test showed there was more microleakage in dentinal margins of composite restorations than in the enamel margins in the test groups (p<0.05). The dentinal margins of the glass ionomer in control groups showed more leakage than the enamel margins, but after the bleaching procedure all experimental groups showed statistically similar microleakage in both the enamel and dentinal margins (p>0.05). CONCLUSION: Postoperative bleaching with carbamide peroxide could increase microleakage in the dentinal margins of composite and the enamel margins of resin-modified glass ionomer restorations. CLINICAL SIGNIFICANCE: Rebonding of resin composite restorations should be considered following bleaching with 15% carbamide peroxide in order to reseal the margins. Resin-modified glass ionomer is not suitable as a filling material before bleaching because of its susceptibility to increased microleakage. PMID- 20020065 TI - Proteome-based analysis of serologically defined tumor-associated antigens in cutaneous lymphoma. AB - Information on specificities of serological responses against tumor cells in cutaneous lymphoma patients is relatively restricted. To advance the knowledge of serological immune responses against and to assess the scope of tumor antigenicity of cutaneous lymphoma, 1- and 2-dimensional Western blot analyses with sera from patients were combined with proteomics-based protein identification. Testing sera from 87 cutaneous lymphoma patients by 1-dimensional Western blot analysis, 64 cases of seroreactivity against lymphoma cells were found. The positive responses were relatively weak, restricted to few antigens in each case, and heterogeneous. To identify the antigens, proteins of the mycosis fungoides cell line MyLa and primary tumor cells were separated by 2-dimensional gel electrophoresis, Western-blotted and probed with heterogeneous and autologous patient sera. The antigens were identified from silver-stained replica gels by MALDI-TOF mass spectrometry. 14 different antigens were assigned and identified with this proteome-serological approach. Only one, vimentin, had been reported before, the other 13 are new antigens for cutaneous lymphomas. PMID- 20020077 TI - The effect of the surface roughness of porcelain on the adhesion of oral Streptococcus mutans. AB - AIM: Dental plaque has a harmful influence on periodontal tissue. When a porcelain restoration is fabricated and refinishing of the glazed surface is inevitable, the increase in surface roughness facilitates the adhesion of plaque and its components. The aim of this in vitro study was to evaluate the effect of surface roughness of glazed or polished porcelain on the adhesion of oral Streptococcus mutans. METHODS AND MATERIALS: A total of 80 metal-ceramic specimens were prepared in the form of disks from two porcelain materials and divided into four groups according to the method of surface finishing. Surface roughness values (Ra-microm) for all specimens were recorded using a profilometer. S. mutans bacteria were isolated from saliva and all specimens were inoculated in test tubes containing a bacterial suspension allowing adhesion of the microorganisms to the specimens to occur. After incubation for 24 hours at 37 degrees C, the specimens were transferred to a sterile saline solution and an inoculum of 0.1 ml from each selected dilution was spread on the selective medium, mitis salivarius bacitracin agar (MSB). Bacterial counts, expressed in colony forming unit (CFU) taking into consideration the dilution factor, were recorded. RESULTS: There was significant correlation (p<0.05) between surface roughness values (Ra-microm) and the amount of bacterial adhesion (CFU x 10(3)). The glazed surface was the smoothest and exhibited the least amount of bacterial adhesion. CONCLUSION: A positive correlation between surface roughness and the amount of S. mutans adhesion was observed. The glazed porcelain surface was considered more biocompatible than other methods of porcelain surface finishing. CLINICAL SIGNIFICANCE: Chairside adjustments of the cervical contour or occlusal surface of porcelain restorations are sometimes necessary before or after cementation. Ideally, an uncemented restoration should be returned to the laboratory for reglazing after all adjustments have been completed. It is important to evaluate various polishing procedures used for these adjusted surfaces to achieve a finished surface that as closely as possible approximates the quality of glazed porcelain. PMID- 20020078 TI - Adhesive bond strengths using self- and light-cured composites. AB - AIM: To evaluate the microtensile bond strength (microTBS) of four adhesive systems to dentin, using self- and light-cured resin composites. METHODS AND MATERIALS: Crowns of human molars were separated from the roots, and the occlusal surfaces were ground to obtain flat superficial dentin. Three etch-and-rinse adhesives--All-Bond 2, One-Step Plus, and OptiBond Solo Plus--and one self etching primer system, Peak SE, were evaluated. Each adhesive group was divided into two subgroups according to the type of resin composite used. A self-cured (Bisfil 2B) or light-cured (Filtek Supreme Plus) resin composite build-up was incrementally inserted to the dentin after each adhesive system was applied. The bonded specimens were stored in water for 24 h and sectioned into beams. Microtensile testing was done, and the data were subjected to ANOVA and Fisher's PLSD test. RESULTS: The microTBS of All-Bond 2 and One-Step Plus was not affected by the type of resin composite used (p=0.3131 and p=0.1562, respectively). The microTBS of OptiBond Solo Plus was significantly reduced when used with self cured resin composite (p<0.0001). Peak SE formed no bond of self-cured resin composite to dentin. CONCLUSION: Some adhesives do not effectively bond self cured resin composite to dentin. CLINICAL SIGNIFICANCE: Incompatibility between adhesives with low pH and certain self-cured resin composites can cause clinical debonding of restorations. PMID- 20020079 TI - Effectiveness of a mouthwash containing Triclosan and Gantrez in the reduction of biofilm and gingivitis: a clinical pilot study. AB - AIM: The aim of this pilot study was to evaluate the effectiveness of a mouthwash containing Triclosan and Gantrez in the reduction of bacterial biofilm and gingivitis. METHODS AND MATERIALS: Thirty patients were assigned to control and experimental groups. The visible plaque index and the marginal bleeding index were determined at the baseline, and at the second and third months, at which time a percentage of bacterial plaque and bleeding was obtained for each patient. All patients were given unlabeled toothpaste without Triclosan and Gantrez and soft-bristle toothbrushes. Additionally, the experimental group patients used unlabeled mouthwash with Triclosan and Gantrez. The participants were instructed to rinse for 60 seconds after brushing, twice a day. RESULTS: At the baseline, the control and experimental group's bacterial plaque and marginal bleeding indices were close to each other (46.61% and 50.75% respectively; 7.62% and 9.59% respectively). At the third month, the experimental group presented bacterial plaque and marginal bleeding indices of 6.56% and 0.43% respectively, while the control presented 11.37% and 2.41% respectively. There were no significant differences between the two groups (p>0.05), with a discrete tendency to better results in the experimental group for both variables. However, there were significant differences among the experimental periods (p<0.05), except between the second and third months of observation. CONCLUSION: The mouthwash did not bring forth an additional advantage when combined with brushing. Additional studies where the observation period may be prolonged and the number of subjects increased are recommended. CLINICAL SIGNIFICANCE: It is important to understand the role chemical control of supragingival plaque can play as an aid in the treatment of periodontal diseases. Since bacterial biofilm is the primary etiological factor in gingivitis and periodontitis, it is essential to know how to control it in order to provide a more complete therapeutic strategy for patient care. PMID- 20020080 TI - Maxillary sinus findings in the elderly: a panoramic radiographic study. AB - AIM: The aim of the present study was to investigate the prevalence of maxillary sinus findings in elderly subjects above the age of 50 through the use of panoramic radiography and to test the hypothesis that such findings are more prevalent in dentate subjects. METHODS AND MATERIALS: A total of 105 subjects over the age of 50 years were examined both clinically and radiographically in the Department of Oral Medicine and Radiology of the Manipal College of Dental Sciences in Manipal, India, over a period of three months for various reasons. Eight were excluded due to the lack of diagnostic quality of their radiographs. The manually calculated kappa test was used to determine the statistical significance of intra-examiner reliability. The other data were analyzed using a manually calculated z-test for proportions where z > or = 1.96 or p < 0.05 were considered statistically significant. RESULTS: The prevalence of radiographic maxillary sinus findings in 97 elderly subjects (aged above 50 years) were investigated with the use of panoramic radiography. Fifty subjects had at least one natural tooth radiographically in the upper jaw and 39 subjects were radiographically edentulous in the upper jaw. Mucous cysts or diffused mucosal thickenings were found in 42.26% of the subjects. Of the mucosal thickenings, 74.35% were found in subjects with a dentate upper jaw (p < 0.05). The prevalence of a mucous cyst was 2.06% in subjects with a dentate upper jaw and a finding of no mucous cysts in edentulous subjects suggests an odontogenic cause. CONCLUSION: According to the findings of the present study, the presence of mucosal thickening or mucosal cysts may be due to the presence of irritative stimuli, often an infection of dental origin, as these findings are more prevalent in dentulous subjects. CLINICAL SIGNIFICANCE: The panoramic radiograph is an excellent diagnostic tool for the identification of mucosal thickening and mucosal cysts. The accurate diagnosis of infective foci (chronic apical periodontitis, deep pockets caused by periodontitis) is necessary, because in conditions of decreased host resistance it may lead to acute sinusitis. PMID- 20020081 TI - Relationship between patients' oral health-related quality of life, satisfaction with dentition, and personality profiles. AB - AIM: The aim of this study was to investigate the relationship between patients' oral health-related quality of life, satisfaction with their dentition, and their personality profiles. METHODS AND MATERIALS: Eighty-four patients (30 males and 54 females; mean age 31.9+/-12.7 years) seeking routine dental treatment were recruited for this study. A "Dental Impact on Daily Living" (DIDL) questionnaire was used to assess dental satisfaction and impact on daily living. An "Oral Health Impact Profile" (OHIP) was used to measure self-reported discomfort, disability, and dysfunction caused by oral conditions. Oral health-related quality of life was assessed using the "United Kingdom Oral Health Related Quality of Life" measure (OHQoL-U.K). A "NEO Five Factor inventory" (NEO-FFI) was used to assess personality profiles. RESULTS: The dentition has a measurable impact on daily living as well as with satisfaction with appearance, pain levels, oral comfort, general performance, and eating capability (p=0.000). Younger patients had more profound oral health impacts (p=0.045) and higher neuroticism scores (0.043). OHIP scores were significantly related to OHQoL-UK scores (p=0.000). DIDL scores had significant correlations with OHIP and OHQoL-UK scores (p<0.05). Significant correlations were established between neuroticism and satisfaction with oral comfort, extraversion and total satisfaction and satisfaction with general performance, and openness and satisfaction with appearance (p<0.05). OHIP and OHQoL-UK scores had no significant correlations with psychological profiles. CONCLUSION: The status of the oral cavity can have a definitive impact on patients' daily living and quality of life regardless age, gender, and level of education. Patients' satisfaction with their dentition has definitive impacts on daily living, quality of life, and dental perceptions. Personality profiles (neuroticism; extraversion, and openness) may influence dental perceptions, play a significant role in shaping satisfaction with dentition, and help with the prediction of the dental impact on daily living. CLINICAL SIGNIFICANCE: Since patients' satisfaction with their dentitions impacts their daily living and quality of life while affected by their psychological profiles, this should be considered when formulating a treatment plan for management for their dentition in order to obtain patient acceptance of the proposed treatment. PMID- 20020082 TI - Effect of the quality of removable prostheses on patient satisfaction. AB - AIM: The objective of this study was to evaluate patient satisfaction with removable prostheses and its relationship with denture quality. METHODS AND MATERIALS: Sixty-seven patients who received 119 new removable prostheses over a period of one year at the College of Dentistry, King Saud University, were recalled for an interview with regard to their satisfaction with their dentures and a clinical examination. Prosthetic quality was determined using the California Dental Association criteria. RESULTS: Results showed 75% and 66% of upper and lower dentures, respectively, were reported by patients as satisfactory. Of all the upper prostheses rated as being of acceptable quality, 94% were also reported by patients as satisfactory, but of those of unacceptable quality, only 52% were reported by patients as unsatisfactory. The corresponding figures for lower dentures were 91% and 71%, respectively. No significant associations were found between patient satisfaction and age, or denture experience. CONCLUSION: Although acceptable quality of removable prostheses usually resulted in patient satisfaction, the finding that some patients were satisfied with their prostheses despite unacceptable quality suggests other factors besides quality affect outcomes. CLINICAL SIGNIFICANCE: Patient satisfaction with removable dentures cannot be fully predicted from only the quality standards of those dentures. PMID- 20020083 TI - Complications of tongue piercing: a review of the literature and three case reports. AB - AIM: The aims of this review of the literature are to offer further insights into possible problems related to tongue piercing and present three case reports showing undesired effects of tongue piercing. BACKGROUND: From a dental perspective, oral piercings and especially tongue piercings are not a harmless fashion trend since they can be associated with local and systemic risks and complications. A search of the literature was conducted to investigate the documentation of health risks associated with tongue piercing using the MEDLINE database as well as the German literature. REVIEW RESULTS: The literature contains numerous case reports, a limited number of studies, and a review describing a wide variety of complications, especially in patients who have undergone tongue piercing. In the majority of cases, the piercing of the tongue is performed in the midline. Piercings are made of different materials, usually metal or synthetic materials. Complications during piercing, immediately following piercing, as well as long term were found. CONCLUSION: The three cases presented here demonstrate some of those adverse effects. The most commonly described oral complication is the damage of teeth and the periodontium caused by tongue piercings. Tongue piercing is a personal decision, but it is important that patients are fully aware of possible oral health hazards. CLINICAL SIGNIFICANCE: Patients need better information on the potential complications associated with tongue piercing. Dental practitioners should educate patients about potential side effects and possible oral, dental, and systemic complications. PMID- 20020084 TI - Aggressive pregnancy tumor mimicking a malignant neoplasm: a case report. AB - AIM: The aim of this report is to present the management of an aggressive, highly proliferative pregnancy tumor with clinical and radiographic characteristics highly suggestive of a malignant neoplasm. BACKGROUND: Pregnancy tumor is a benign hyperplastic gingival lesion occurring during pregnancy that is indistinguishable from a pyogenic granuloma arising in nonpregnant females, or in males. The lesion usually grows over a few months and tends to bleed. CASE DESCRIPTION: A 28-year-old woman at four months of gestation was referred for a massive gingival swelling (5.5 cm in greatest diameter) on the mandibular left side. The lesion was painful and continued to grow very rapidly over a three-week period, with spontaneous bleeding, and it interfered with speech and mastication. Advanced alveolar bone loss also was found beneath the lesion. A malignant process was suspected, and an incisional biopsy revealed a pregnancy tumor. The lesion was excised under general anesthesia during the pregnancy with no untoward reactions. SUMMARY: Pregnancy tumor represents an important differential diagnosis of oral masses and can behave in a very aggressive fashion, mimicking a malignant tumor. CLINICAL SIGNIFICANCE: This lesion should always be included in the differential diagnosis of soft tissue masses in a pregnant woman even if the lesion is clinically very aggressive. It is acceptable practice to excise aggressive variants of this lesion during pregnancy to avoid distressing side effects. PMID- 20020085 TI - The restoration of function and esthetics of a patient with amelogenesis imperfecta using a combination of orthodontic and prosthodontic treatment: a case report. AB - AIM: The purpose of this case report is to present the esthetic and functional rehabilitation of the teeth in a 20-year-old patient with amelogenesis imperfecta (AI), facial asymmetry due to functional mandibular shift, and unilateral posterior crossbite. BACKGROUND: AI is a group of hereditary defects of enamel unassociated with any other generalized defects. AI results in poor development or the complete absence of the enamel of the teeth caused by improper differentiation of ameloblasts. CASE DESCRIPTION: This report describes the diagnosis and treatment of a young female patient with AI and facial asymmetry using a combined orthodontic-prosthodontic approach. Initially, the posterior crossbite, mandibular shift, and facial asymmetry were treated orthodontically. Later, metal-ceramic crowns for posterior teeth and all-ceramic crowns for anterior teeth were fabricated for final restorations. SUMMARY: Coordinated orthodontic and prosthodontic treatment, with careful consideration of patient expectations and requests, were critical for a successful outcome and patient satisfaction. CLINICAL SIGNIFICANCE: The complexity of the management of patients with AI supports the suggestion that the dental profession should have appropriate methods for the rehabilitation of rare dental disorders. PMID- 20020086 TI - A new concept in restorative dentistry: light-induced fluorescence evaluator for diagnosis and treatment. Part 1: Diagnosis and treatment of initial occlusal caries. AB - AIM: The objective of this in vivo experiment is to propose an innovative therapeutic concept using a light-induced fluorescence evaluator for diagnosis and treatment (LIFEDT) that is based on the imaging and autofluorescence of dental tissues. BACKGROUND: Processes with the aim of diagnosing carious lesions in the initial stage with optimum sensitivity and specificity employ a wide variety of technologies, but like the conventional diagnosis tools, they remain either inefficient or too subjective. TECHNIQUE: This experiment evaluated a fluorescence light-induced camera that illuminates tooth surfaces within an excitation radiation band of light with a wavelength of 450 nm and facilitates a high magnification image. CONCLUSION: An analysis of 50 occlusal grooves revealed three clinical forms of enamel caries: (1) enamel caries on the surface, (2) suspicious grooves with a positive autofluorescent red signal, and (3) suspicious grooves with a neutral fluorescent dark signal. Two decision-making diagrams were proposed in accordance with international recommendations for preventive dentistry, but modified as a result of the accurate information obtained with this new LIFEDT device. CLINICAL SIGNIFICANCE: The lighting of suspect occlusal grooves with the SoproLife camera enables observation of any variations in the optical properties to refine a caries diagnosis and facilitates more than a 50x magnification of occlusal groove anatomy to provide additional information on the carious potential of the tooth surface. PMID- 20020087 TI - AAOMP case challenge: A mass of the maxillary tuberosity. AB - A 48-year-old African-American female was evaluated for an asymptomatic bony expansion of the right maxillary tuberosity. PMID- 20020088 TI - An endobronchial lipoma mimicking asthma and malignancy. AB - Endobronchial lipomas are rare benign tumours of the lung. Bronchial occlusion may lead to a misdiagnosis of asthma or malignancy. We describe a 52-year-old man treated for asthma for several years, who presented with non-resolving right upper lobe pneumonia. Bronchoscopy proved to be diagnostic and therapeutic. Clinical characteristics of this unique entity are discussed. PMID- 20020089 TI - [Physician's advice and treatment of smokers]. AB - This article describes the health effects of smoking and smoking cessation. The doctor's office provides advantages for the counseling and treatment of smokers, such as continuous patient contacts, the option to address smoking in the context of smoking-related diseases, ubiquitous and low-threshold access to health care, therefore, offering equal access to lower socioeconomic classes. However, barriers exist for the implementation of smoking-related counseling in private practice due to the lack of reimbursement, insufficient qualification, lack of available time as well as lack of patients' motivation. Moreover, in the German healthcare system tobacco dependency has not yet been acknowledged as a treatment demanding disease. Existing obstacles should be overcome by adjustment of reimbursement schemes for private practice and hospitals as well as reimbursement for smoking cessation medication proven to be effective for tobacco-dependent smokers. The German Medical Association has established a training program for doctors who want to become active in smoking cessation and has developed printed material for the counseling of smokers in private practice. PMID- 20020090 TI - [Clinical studies outside university clinics : What are the problems to implement this?]. AB - The demand for high quality evidence-based surgical treatment in Germany and awareness of the poor quality of surgical trials highlight the basic necessity of randomized controlled trials. In six surgical trial centers a professional infrastructure for surgical trials is in the process of being established since 2006.The aim is the initiation of surgical multicenter trials which can be effectively conducted by local networking. To accomplish a timely recruitment of patients it is necessary to integrate surgical departments outside university hospitals into multicenter trials. With a questionnaire survey of non-university surgical departments in the federal states of Berlin and Brandenburg, interest, experience in clinical trials and structural conditions in these departments were evaluated. Based on the results of this survey the possibilities to integrate non university surgical departments into multicenter trials and how a high recruitment of patients can be motivated will be discussed in this article. PMID- 20020092 TI - [Complicated intra-abdominal infections: pathogens, resistance. Recommendations of the Infectliga on antbiotic therapy]. AB - Intra-abdominal infections are generally the result of invasion and multiplication of enteric bacteria in the wall of a hollow viscus within the abdomen to produce peritonitis or abscess. When the infection extends into the peritoneal cavity or another normally sterile region of the abdominal cavity, the infection is described as a "complicated" intra-abdominal infection. Treatment of patients with complicated intra-abdominal infections involves antimicrobial therapy, generally in conjunction with an appropriate and timely surgical source control. Nearly all intra-abdominal infections are caused by multiple microorganisms that constitute the intestinal flora (aerobes and facultative and obligate anaerobes, with Enterobacteriaceae, enterococci and Bacteroides fragilis isolated most frequently). The emergence of drug resistance (e.g. ESBL-producing Enterobacteriaceae or resistant enterococci and staphylococci) poses a substantial threat to patients with surgical infections. Especially in patients with nosocomially acquired infections inadequate empiric antibiotic treatment is associated with treatment failure and death. In patients at risk broader spectrum antibiotic regimens with coverage of resistant Gram-negative bacilli and anaerobes and Gram-positive bacteria such as enterococci (including VRE) and staphylococci should be considered. PMID- 20020093 TI - [Perioperative management of interventions on the musculoskeletal system]. PMID- 20020091 TI - [Survival, mortality and quality of life after pylorus-preserving or classical Whipple operation. A systematic review with meta-analysis]. AB - BACKGROUND: Two surgical procedures are mainly performed for the treatment of pancreatic head cancer and periampullary carcinoma: the classical Whipple operation and the pylorus-preserving Whipple operation. METHODS: This manuscript represents an extension of a systematic review and meta-analysis previously published in the Annals of Surgery. A systematic literature search was performed in MEDLINE, EMBASE and the Cochrane Library (central) to identify randomized controlled trials (RCTs) and observational studies. A meta-analysis based on a random-effects model was performed for the hazard ratios (HR) of survival and the odds ratios (OR) of postoperative mortality. The results of the different studies on quality of life (QoL) could not be summarized quantitatively in a meta analysis and were therefore summarized qualitatively. Subgroup analyses were performed by study type, RCTs, prospective cohort studies (PSs), retrospective cohort studies (RSs), study quality and tumor localization (pancreatic head cancer versus periampullary carcinoma). RESULTS: The systematic literature search retrieved 4,503 studies of which 4,460 did not fulfill the inclusion criteria. The remaining 43 studies (6 RCTs, 12 PSs and 25 RSs) representing 3,893 patients were finally included in the review. There was neither a significant survival difference for patients with pancreatic head cancer in the pooled estimate of the RCTs (HR 0.80; 95% CI 0.53-1.22; p=0.16) nor in the pooled estimate of the PSs (HR 0.84; 95% CI 0.7-1.0; p=0.95) or the RSs (HR 0.84; 95% CI 0.7-1.01; p=0.21). Survival of patients with periampullary carcinoma was not significantly different in the RCTs (HR 1.02; 95% CI 0.49-2.13; p=0.3), the PSs (HR 1.26; 95% CI 0.46 3.42; p=0.65) or the RSs (HR 0.86; 95% CI 0.6-1.24; p=0.33). Postoperative mortality was not significantly different after both types of operations (RCTs: HR 0.49; 95% CI 0.17-1.4; p=0.18; PSs: HR 0.63; 95% CI 0.34-1.18; p=0.15; RSs: HR 0.7; 95% CI 0.37-1.31; p=0.27). QoL was reported as either the same in both groups or in favor of the pylorus-preserving Whipple operation. CONCLUSIONS: Mortality, survival and QoL were not significantly different between the classical Whipple and the pylorus-preserving Whipple operations. Given the poor quality of the underlying trials a pragmatic RCT is recommended to prove the findings of this systematic review. PMID- 20020094 TI - [Fractures of the lateral clavicle]. AB - Fractures of the lateral clavicle end account for 12-15 percent of all clavicle fractures. In contrast to the clear treatment of midshaft fractures the therapy of the distal third is still open to controversy. The high non-union rate up to 40 percent that occurs with the lateral end fractures shows the special biomechanical mechanisms. The role of the coracoclavicular ligaments is comparable to the acromioclavicular joint disruption. To make a clear decision about the therapy we need a treatment based classification such as the one by Jager and Breitner, which distinguishes four different fracture types. The treatment options reach from conservative therapy up to numerous different operative techniques. The aim of this article is to demonstrate a simple and save way for the diagnosis and therapy of that special kind of fracture. PMID- 20020095 TI - [Inadmissible cooperation contract between hospital and physician. New court decision confirms criticism of the previous practice]. AB - The decision of the Higher Regional Court of Dusseldorf from 01.09.2009 (20 U 121/08), given below in parts, confirms the decision of the Regional Court Duisburg from 01.04.2008 (4 O 300/07) published in Der Urologe issue 10.2008 (1355 ff). The hope expressed in the literature (e.g. Wienke/Janke, OM 4-2009, 461) that the decision of the Regional Court Duisburg would not be upheld on appeal, proved to be unwarranted. PMID- 20020096 TI - Enhancement of glucagon secretion in mouse and human pancreatic alpha cells by protein kinase C (PKC) involves intracellular trafficking of PKCalpha and PKCdelta. AB - AIMS/HYPOTHESIS: Protein kinase C (PKC) regulates exocytosis in various secretory cells. Here we studied intracellular translocation of the PKC isoenzymes PKCalpha and PKCdelta, and investigated how activation of PKC influences glucagon secretion in mouse and human pancreatic alpha cells. METHODS: Glucagon release from intact islets was measured in static incubations, and the amounts released were determined by RIA. Exocytosis was monitored as increases in membrane capacitance using the patch-clamp technique. The expression of genes encoding PKC isoforms was analysed by real-time PCR. Intracellular PKC distribution was assessed by confocal microscopy. RESULTS: The PKC activator phorbol 12-myristate 13-acetate (PMA) stimulated glucagon secretion from mouse and human islets about fivefold (p < 0.01). This stimulation was abolished by the PKC inhibitor bisindolylmaleimide (BIM). Whereas PMA potentiated exocytosis more than threefold (p < 0.001), BIM inhibited alpha cell exocytosis by 60% (p < 0.05). In mouse islets, the PKC isoenzymes, PKCalpha and PKCbeta1, were highly abundant, while in human islets PKCeta, PKCepsilon and PKCzeta were the dominant variants. PMA stimulation of human alpha cells correlated with the translocation of PKCalpha and PKCdelta from the cytosol to the cell periphery. In the mouse alpha cells, PKCdelta was similarly affected by PMA, whereas PKCalpha was already present at the cell membrane in the absence of PMA. This association of PKCalpha in alpha cells was principally dependent on Ca(2+) influx through the L-type Ca(2+) channel. CONCLUSIONS/INTERPRETATION: PKC activation augments glucagon secretion in mouse and human alpha cells. This effect involves translocation of PKCalpha and PKCdelta to the plasma membrane, culminating in increased Ca(2+)-dependent exocytosis. In addition, we demonstrated that PKCalpha translocation and exocytosis exhibit differential Ca(2+) channel dependence. PMID- 20020097 TI - Long-term detrimental consequences of the onset of type 1 diabetes on annual earnings--evidence from annual registry data in 1990-2005. AB - AIMS/HYPOTHESIS: Young adults in the early stages of their participation in the labour market may be particularly vulnerable to the effects of onset of a chronic disease. Our aim was to quantify the consequences of the onset of type 1 diabetes in young adults on annual earnings, using individual-level longitudinal data before and after the onset of diabetes. METHODS: The Econ-DISS database contains annual socioeconomic information for 1990-2005 from Statistics Sweden. Econ-DISS includes data for persons with diabetes onset at the age of 15-34 years between 1983 and 2005, registered in the national Diabetes Incidence Study in Sweden (DISS) database, and for controls. Considering the onset of type 1 diabetes as an unanticipated and significant life event, we compared the progression of annual earnings for 3,650 cases born between 1949 and 1970 before and after onset of diabetes with that of 14,629 controls. Possible confounders--education, participation in the labour market, sick leave and parental education--were analysed. RESULTS: We found no differences between the groups in annual earnings or participation in the labour market before onset of diabetes. After onset, persons with type 1 diabetes gradually lagged behind the controls. Their median annual earnings were lower in each year from 1995 to 2005 (p < 0.01). The difference in 2005 was euro (EUR) 1,411 (5.3%). Controlling for confounders, duration of type 1 diabetes > or = 10 years was associated with 4.2% (men) and 8.1% (women) lower average annual earnings for persons with upper secondary education only who were active in the labour market. CONCLUSION/INTERPRETATION: The onset of type 1 diabetes in young adults has long-term detrimental consequences on earnings that cannot be attributed to confounders. PMID- 20020098 TI - Medial approaches to osteochondral lesion of the talus without medial malleolar osteotomy. AB - The purpose of this study was to determine the area of the talus that can be reached through combined anterior and posterior arthrotomy without medial malleolar osteotomy. Five fresh-frozen cadaver foot-ankle specimens were examined using posteromedial approach and anteromedial approach. We calculated the size of the marked area beginning from the posteromedial corner of the talus in the posteromedial approach and beginning from the anteromedial corner in the anteromedial approach. From the posteromedial talus, we can access 33% of the talus' AP length and 30% of its medial to lateral length through a posteromedial approach. From the anteromedial arthrotomy, 50% of the AP length and 31% of the medial to lateral length can be reached. This leaves approximately 20% that is not accessible. If the osteochondral lesion is within the accessible area through either a posteromedial or anteromedial approach as viewed on MRI/CT, it can be safely reached without a medial malleolar osteotomy. PMID- 20020099 TI - Snapping biceps femoris tendon treated with an anatomic repair. AB - Three patients were seen for pain and snapping over the lateral aspect of their symptomatic knee during deep knee flexion. On physical examination, each patient had subluxation of the long head of the biceps over the lateral aspect of the fibular head. Each patient underwent an anatomic repair of the torn anterior arms of the short and long biceps femoris to their anatomic insertion sites with suture anchors. All patients had normal return of function without pain or further subluxation events. PMID- 20020100 TI - Chondrogenesis in a hyaluronic acid scaffold: comparison between chondrocytes and MSC from bone marrow and adipose tissue. AB - Treatment of focal lesions of the articular cartilage of the knee using chondrocytes in a hyaluronic acid (HA) scaffold is already being investigated in clinical trials. An alternative may be to use mesenchymal stem cells (MSC). We have compared articular chondrocytes with MSC from human bone marrow (BM) and adipose tissue (AT), all cultured in HA scaffolds, for their ability to express genes and synthesize proteins associated with chondrogenesis. The cells were expanded in monolayer cultures. After seeding into the scaffold, the chondrocytes were maintained in medium, while the two MSC populations were given a chondrogenic differentiation medium. Chondrogenesis was assessed by real-time RT PCR for chondrocyte-associated genes, by immunohistochemistry and by ELISA for collagens in the supernatant. Redifferentiation of the dedifferentiated chondrocytes in the HA scaffold was shown by a modest increase in type II collagen mRNA (COL2A1) and reduction in COL1A1. BM-MSC expressed 600-fold higher levels of COL2A1 than chondrocytes after 3 weeks in the scaffold. The levels of aggrecan (AGC1) and COL1A1 were similar for chondrocyte and BM-MSC scaffold cultures, while COL10A1 was higher in the BM-MSC. AT-MSC expressed levels of COL2A1 and COL1A1 similar to chondrocytes, but less AGC1 and COL10A1. Surprisingly, little collagen II protein was observed in the scaffold. Instead, collagen II was found in the culture medium. Chondrogenesis in HA scaffolds was more efficient using BM-MSC than AT-MSC or chondrocytes. Some of the secreted collagen II escaped entrapment in the extracellular space and was detected in the culture medium. PMID- 20020101 TI - Rotational laxity greater in patients with contralateral anterior cruciate ligament injury than healthy volunteers. AB - Rotational stability of the knee has been traditionally difficult to quantify, limiting the ability of the orthopedic community to determine the potential role of rotational laxity in the etiology of anterior cruciate ligament (ACL) injuries. The purposes of this multicenter cohort study were to evaluate the reliability of a robotic axial rotation measurement system, determine whether the uninjured knees of patients that had previous contralateral ACL reconstruction demonstrated different rotational biomechanical characteristics than a group of healthy volunteers, and determine whether knee rotational biomechanical characteristics differ between male and female non-injured limbs in groups of both healthy volunteers and patients with a previous contralateral ACL injury. Fourteen healthy volunteers and 79 patients with previous unilateral ACL injury participated in this study. Patients were tested using a computerized tibial axial rotation system. Only the normal (non-operated) knee data were used for analysis. In order to assess the reliability of the robotic measurement system, 10 healthy volunteers were tested daily over four consecutive days by four different examiners. Rotational laxity and compliance measures demonstrated excellent reliability (ICC = 0.97). Patients with a contralateral ACL injury demonstrated significantly increased tibial internal rotation (20.6 degrees vs. 11.4 degrees , P < 0.001) and reduced external rotation (16.7 degrees vs. 26.6 degrees , P < 0.001) compared to healthy volunteers. Females demonstrated significantly increased internal and external rotation, as well as significantly increased rotational compliance compared with males (P < 0.05). Computer-assisted measurement techniques may offer clinicians an accurate, reliable, non-invasive method to select the most appropriate preventative or surgical interventions for patients with increased knee rotational laxity. PMID- 20020102 TI - Mutation in the lysA gene impairs the symbiotic properties of Mesorhizobium ciceri. AB - A Tn5-induced mutant of Mesorhizobium ciceri, TL28, requiring the amino acid lysine for growth on minimal medium was isolated and characterized. The Tn5 insertion in the mutant strain TL28 was located on a 6.8-kb EcoRI fragment of the chromosomal DNA. Complementation analysis with cloned DNA indicated that 1.269 kb of DNA of the 6.8-kb EcoRI fragment restored the wild-type phenotype of the lysine-requiring mutant. This region was further characterized by DNA sequence analysis and was shown to contain a coding sequence homologous to lysA gene of different bacteria. The lys (-) mutant TL28 was unable to elicit development of effective nodules on the roots of Cicer arietinum L. There was no detectable level of lysine in the root exudates of chickpea. However, addition of lysine to the plant growth medium restored the ability of the mutant to produce effective nodules with nitrogen fixation ability on the roots of C. arietinum. PMID- 20020103 TI - Association of melamine exposure with urinary stone and oxidative DNA damage in infants. AB - There is evidence in experimental animals for the urolithiasis and carcinogenicity of melamine, but no evidence for melamine in humans. To evaluate any association between melamine-contaminated powdered formula (MCPF) feeding and urolithiasis, and further the MCPF feeding and oxidative damage to DNA in infants. A cross-sectional study was carried out to assess urolithiasis and urinary 8-hydroxy-2'-deoxyguanosine (8-OHdG) in four groups of infants according to the type of feeding: (1) Infants receiving over 90% of their intake as MCPF. (2) Infants receiving 50-90% of their intake as MCPF. (3) Infants receiving less than 50% of their intake as MCPF. (4) Infants receiving over 90% of their intake as imported milk powdered formula free of melamine contamination. Groups 1 to 3 are the observation groups, and Group 4 is the reference group. There is a significant correlation between urolithiasis and percentage of MCPF intake. The mean urinary 8-OHdG concentrations for Groups 1, 2, 3, and 4 were: 2.03 +/- 0.52, 1.67 +/- 0.28, 1.90 +/- 0.39, and 1.85 +/- 0.47 micromoles per mole of creatinine, respectively. There were no significant differences in the mean urinary 8-OHdG concentrations among the observation and control groups. There were also no correlation between mean urinary 8-OHdG excretions and percentage of MCPF intake. Our data suggested that melamine exposure were associated with urolithiasis, but it might not cause any increase in oxidative damage of DNA in infants. PMID- 20020104 TI - Metabolism of arsenic in human liver: the role of membrane transporters. AB - Metabolism of inorganic arsenic (iAs) is one of the key factors determining the character of adverse effects associated with exposure to iAs. Results of previous studies indicate that liver plays a primary role in iAs metabolism. This paper reviews these results and presents new data that link the capacity of human hepatocytes to metabolize iAs to the expression of specific membrane transporters. Here, we examined relationship between the expression of potential arsenic transporters (AQP9, GLUT2, P-gp, MRP1, MRP2, and MRP3) and the production and cellular retention of iAs and its methylated metabolites in primary cultures of human hepatocytes exposed for 24 h to subtoxic concentrations of arsenite. Our results show that the retention of iAs and methylarsenic metabolites (MAs) by hepatocytes exposed to sub-micromolar concentrations of arsenite correlates negatively with MRP2 expression. A positive correlation was found between MRP2 expression and the production of dimethylarsenic metabolites (DMAs), specifically, the concentration of DMAs in culture media. After exposures to high micromolar concentrations of arsenite which almost completely inhibited MAs and DMAs production, a positive correlation was found between the expression of GLUT2 and cellular retention of iAs and MAs. MRP3, AQP9, or P-gp expression had no effect on the production or distribution of iAs, MAs, or DMAs, regardless of the exposure level. Hepatocytes from seven donors used in this study did not contain detectable amounts of MRP1 protein. These data suggest that MRP2 plays an important role in the efflux of DMAs, thus, regulating kinetics of the methylation reactions and accumulation of iAs and MAs by human hepatocytes. The membrane transport of iAs by high-capacity GLUT2 transporters is not a rate limiting step for the metabolism of arsenite at low exposure level, but may play a key role in accumulation of iAs after acute exposures which inhibit iAs methylation. PMID- 20020105 TI - Toxicity of penta- and decabromodiphenyl ethers after repeated administration to rats: a comparative study. AB - Until recently, pentabromodiphenyl (PentaBDE) and decabromodiphenyl (DecaBDE) ethers were commonly used as flame retardants in a wide array of products, mostly in the production of plastics utilized in the electric, electronic and textile industries. The aim of this study was to compare the toxicity of PentaBDE and DecaBDE after their repeated (7-28 days) intragastric administration to rats. The compounds were given at doses of 2, 8, 40 or 200 mg/kg/day (PentaBDE) and 10, 100 or 1,000 mg/kg/day (DecaBDE). The repeated administration of PentaBDE disturbed redox homeostasis, which was manifested by lower total antioxidant status and increased activity of glutathione reductase in serum and higher concentrations of glutathione reduced and malondialdehyde in the liver. The occurrence of these effects was not observed after DecaBDE administration. The results of histopathological examination showed fatty degeneration after administration of the highest dose of PentaBDE. The repeated administration of PentaBDE also caused the increase in relative liver mass, dose-dependent increase in the activity of CYP 1A (EROD) and CYP 2B (PROD), 7-12- and 2-8-fold, respectively, as well as enhanced level of CYP 1A1 (5-30-fold) and CYP 4A (2-4.5-fold). The administration of DecaBDE induced much less pronounced changes: a maximum 2.8-fold increase in the activity of CYP 1A, a twofold increase in CYP 2B, and no alterations in other parameters under study. Contrary to DecaBDE, PentaBDE disturbed redox homeostasis, and induced liver microsomal enzymes. Fatty degeneration in liver caused by this compound was also found. PMID- 20020106 TI - Gene expression profiles in the brain of the neonate mouse perinatally exposed to methylmercury and/or polychlorinated biphenyls. AB - Methylmercury (MeHg) and polychlorinated biphenyls (PCBs) are environmentally persistent neurodevelopmental toxicants. The primary source of human exposure is the consumption of contaminated fish, seafood and marine mammals. However, little is known about the molecular mechanisms of MeHg and PCB toxicities and interactions between these contaminants. We investigated the functional profiles of differently expressed genes in the brains of offspring mice perinatally exposed to MeHg and/or PCBs to elucidate how these contaminants interact with each other. Pregnant mice (C57BL/6) were divided into four groups by exposure: (1) vehicle control, (2) MeHg alone, (3) PCBs alone, (4) MeHg + PCBs. Gene expression analysis of the brains of offspring mice was carried out with 4 x 44 K whole mouse genome's microarrays (Agilent) on postnatal day 1. The gene expression pattern of the MeHg exposure-group differed from that of the PCB exposure group. The MeHg + PCB group expressed a larger number of genes, most of which were not expressed in the MeHg group or PCB group. It was revealed that gene expression was greatly increased, and the most altered genes were found with co-exposure. The genes were related to the functional categories of development, inflammation, calcium ion homeostasis, signal transduction, the ubiquitin proteasome pathway and detoxication. The ubiquitin-proteasome system and detoxication categories might function for protection against the toxicity induced by co-exposure to MeHg and PCBs. These results suggest that co-exposure does not simply exacerbate the toxicity of MeHg alone or PCB alone, but stimulates a protection system. PMID- 20020107 TI - Alcohol breaks down interhemispheric inhibition in females but not in males: alcohol and frontal connectivity. AB - INTRODUCTION: Alcohol has renowned behavioral disinhibitory properties which are suggested to involve reductions in frontal lobe functioning as a result of diminished interhemispheric connectivity. METHODS: To examine sex differences in frontal interhemispheric connectivity in response to alcohol, 12 female and ten male healthy volunteers received a single administration of 0.5 per thousand alcohol in a placebo-controlled counterbalanced crossover design. Paired-pulse transcranial magnetic stimulation was applied to measure transcallosal inhibition (TCI) between the left and right primary motor cortex (M1). RESULTS: Results showed significant reductions in TCI after alcohol administration in female participants exclusively. DISCUSSION: These findings provide the first evidence that moderate doses of alcohol differentially affect frontal interhemispheric connectivity in males and females. The present data may shed new light on the physiological mechanisms underlying sex differences in the susceptibility to alcohol. PMID- 20020108 TI - The effects of adolescent methylphenidate self-administration on responding for a conditioned reward, amphetamine-induced locomotor activity, and neuronal activation. AB - BACKGROUND: Abuse of methylphenidate (Ritalin) is rising, particularly during adolescence and early adulthood, but the long-term effects of its abuse during adolescence are unclear. METHODS: In experiment 1, we examined the effect of adolescent methylphenidate self-administration (0.0625 mg/infusion), as compared with cocaine self-administration (0.125 mg/infusion), under a fixed ratio 1 schedule of reinforcement in male Sprague-Dawley rats during adolescence (postnatal day (PND) 32-47) on adult dopamine-mediated behaviors (PND >70). These included responding for a conditioned reward (CR), a measure of incentive motivation, and amphetamine-induced locomotor activity. In experiment 2, we aimed to replicate and enhance the effects observed in experiment 1, and we also examined the effects of methylphenidate self-administration during adolescence on adult amphetamine-induced zif268 messenger ribonucleic acid (mRNA) expression. RESULTS: Adolescent rats self-administered both cocaine and methylphenidate. There was no effect of adolescent drug self-administration on adult baseline or amphetamine-induced responding for a CR. However, both adolescent methylphenidate and cocaine self-administration increased amphetamine-induced locomotion. Adolescent methylphenidate self-administration also enhanced amphetamine-induced zif268 mRNA expression in the nucleus accumbens. CONCLUSIONS: Our findings suggest that repeated, behaviorally contingent exposure to methylphenidate during adolescence enhances responsivity to the locomotor-stimulating and neuronal activating effects of amphetamine but not incentive motivation. PMID- 20020109 TI - GBR 12909 administration as a mouse model of bipolar disorder mania: mimicking quantitative assessment of manic behavior. AB - RATIONALE: Mania is a core feature of bipolar disorder (BD) that traditionally is assessed using rating scales. Studies using a new human behavioral pattern monitor (BPM) recently demonstrated that manic BD patients exhibit a specific profile of behavior that differs from schizophrenia and is characterized by increased motor activity, increased specific exploration, and perseverative locomotor patterns as assessed by spatial d. OBJECTIVES: It was hypothesized that disrupting dopaminergic homeostasis by inhibiting dopamine transporter (DAT) function would produce a BD mania-like phenotype in mice as assessed by the mouse BPM. METHODS: We compared the spontaneous locomotor and exploratory behavior of C57BL/6J mice treated with the catecholamine transporter inhibitor amphetamine or the selective DAT inhibitor GBR 12909 in the mouse BPM. We also assessed the duration of the effect of GBR 12909 by testing mice in the BPM for 3 h and its potential strain dependency by testing 129/SvJ mice. RESULTS: Amphetamine produced hyperactivity and increased perseverative patterns of locomotion as reflected in reduced spatial d values but reduced exploratory activity in contrast to the increased exploration observed in BD patients. GBR 12909 increased activity and reduced spatial d in combination with increased exploratory behavior, irrespective of inbred strain. These effects persisted for at least 3 h. CONCLUSIONS: Thus, selectively inhibiting the DAT produced a long lasting cross-strain behavioral profile in mice that was consistent with that observed in manic BD patients. These findings support the use of selective DAT inhibition in animal models of the impaired dopaminergic homeostasis putatively involved in the pathophysiology of BD mania. PMID- 20020110 TI - Raman spectroscopic analysis of minerals and organic molecules of relevance to astrobiology. AB - Characteristic geological features and hydrated minerals recently found on the surface of Mars by the NASA planetary rovers Spirit and Opportunity suggest that a possible biosphere could have once existed there. Analytical instrumentation protocols for the unequivocal detection of biomarkers in suitable geological matrices are critical for future unmanned explorations, including the forthcoming ESA-ExoMars mission scheduled for 2018. Raman spectroscopy is currently a part of the Pasteur instrumentation suite of the ExoMars mission scheduled for 2018 for the remote detection of extant or extinct life signatures in the Martian surface and subsurface. Terrestrial analogues of Martian sites have been identified, and the biogeological modifications incurred as a result of extremophilic survival activity have been studied. Polyaromatic hydrocarbons (PAHs) are recognised as a class of degradation product that occur from biological processes terrestrially. In this work, various concentrations of polyaromatic hydrocarbons in matrices of gypsum, calcite and quartz have been investigated by Raman microspectrometry to determine the lowest detectable organic levels. The studies are conceived in simulation of their potential PAHs identification in geobiological conditions in Martian scenarios. Two laser source wavelengths, namely, 785 and 633 nm, were adopted to excite Raman spectra from the PAHs, which represent degraded carbons and therefore potentially provide a key bimolecular marker of ancient life. PMID- 20020111 TI - Host-molecule-coated quantum dots as fluorescent sensors. AB - "Host" molecules, containing a binding site that is highly specific for an analyte "guest," are used as sensors to register analyte binding through a variety of mechanisms such as colorimetric, fluorescent, or electrochemical signals. There is increasing interest in the host-guest chemistry on the surface of quantum dots (QDs) and in the changes that it produces in the luminescent properties of QDs. The bulk of this study focuses on those QDs with bound host molecules (crown ether, cyclodextrin, calixarene, and porphyrin) and the selectivity they display toward metal ions and small organic molecules. PMID- 20020112 TI - Application of electrospray ionization hybrid ion trap/time-of-flight mass spectrometry in the rapid characterization of quinocetone metabolites formed in vitro. AB - The application of electrospray ionization hybrid ion trap/time-of-flight mass spectrometry coupled with high-performance liquid chromatography (LC/MS-IT-TOF) in the rapid characterization of in vitro metabolites of quinocetone was developed. Metabolites formed in rat liver microsomes were separated using a VP ODS column with gradient elution. Multiple scans of metabolites in MS and MS(2) modes and accurate mass measurements were automatically performed simultaneously through data-dependent acquisition in only a 30-min analysis. Most measured mass errors were less than 10 ppm for both protonated molecules and fragment ions using external mass calibration. The elemental compositions of all fragment ions of quinocetone and its metabolites could be rapidly assigned based upon the known compositional elements of protonated molecules. The structure of metabolites were elucidated based on the combination of three techniques: agreement between their proposed structure, the accurate masses, and the elemental composition of ions in their mass spectra; comparison of their changes in accurate molecular masses and fragment ions with those of parent drug or metabolite; and the elemental compositions of lost mass numbers in proposed fragmentation pathways. Twenty seven phase I metabolites were identified as 11 reduction metabolites, three direct hydroxylation metabolites, and 13 metabolites with a combination of reduction and hydroxylation. All metabolites except the N-oxide reduction metabolite M6 are new metabolites of quinocetone, which were not previously reported. The ability to conduct expected biotransformation profiling via tandem mass spectrometry coupled with accurate mass measurement, all in a single experimental run, is one of the most attractive features of this methodology. The results demonstrate the use of LC/MS-IT-TOF approach appears to be rapid, efficient, and reliable in structural characterization of drug metabolites. PMID- 20020113 TI - Cortico-geniculate feedback linking the visual fields surrounding the blind spot in the cat. AB - Previous studies have shown that neurons in layer 6 of V1 are activated by visual stimuli that induce perceptual filling-in at the blind spot (BS). As the main target of layer 6 neurons is the lateral geniculate nucleus (LGN), we speculate that the cortico-geniculate projection is involved in mediating filling-in at BS. To begin to test that hypothesis, we examined whether there is an anatomical basis for integration of visual signals from both sides of BS by cortico geniculate feedback neurons in V1. We injected an anterograde tracer into a site adjacent to the region representing BS. We observed that numerous axons traverse the neuron-free gap that retinotopically corresponds to BS within LGN. This indicates that visual signals from one side of BS are conveyed to the opposite side via a feedback connection. Cortico-geniculate feedback projection may integrate visual signals from around BS and contribute to perceptual filling-in at BS. PMID- 20020114 TI - A flexible sequential learning deficit in patients with Parkinson's disease: a 2 x 8 button-press task. AB - A 2 x 8 button-press task is a sequential hand movement task in which subjects are required to press eight pairs of buttons as accurately and quickly as possible. The 2 x 8 task allows us to examine flexible sequential learning, more aptly called sequence-unselective learning. Sequence-unselective learning is observed after repeated experiences with the task, when subjects have shown good progress in learning, with new sequences as well as previously learned ones. Although cognitive inflexibility has been reported in patients with Parkinson's disease (PD), there have been few studies investigating their flexibility in sequential learning. We examined PD patients' ability for sequence-unselective learning through the use of a 2 x 8 button-press task. In the first session, PD patients and subjects from the control group performed a sequential 2 x 8 task until the learning criterion was fulfilled (Session 1). After 1 month, they participated in other sessions: one involving the learned sequence (Session 2) and another involving the new sequence (Session 3). We found that PD patients made more errors than the normal control subjects only when learning the new sequence (Session 3) (P < 0.01). In Session 3, control subjects reached the learning target with fewer errors than in the Session 1 (normal sequence unselective learning), whereas the PD patients did not exhibit such an improvement. Our results revealed a sequence-unselective deficit in PD patients. The deficit may help to emphasize the cognitive and physical inflexibility of PD. PMID- 20020115 TI - Altered carotid plaque signal among different repetition times on T1-weighted magnetic resonance plaque imaging with self-navigated radial-scan technique. AB - INTRODUCTION: Magnetic resonance (MR) plaque imaging for carotid arteries is usually performed by using an electrocardiograph (ECG)-gating technique to eliminate pulsation-related artifacts, which can affect the plaque signals because of varied repetition time (TR) among patients. Hence, we investigated whether differences in TR causes signal alterations of the carotid plaque by using a non-gated plaque imaging technique. METHODS: We prospectively examined 19 patients with carotid stenosis by using a T1-weighted self-navigated radial-scan technique with TRs of 500, 700, and 900 ms. The signal intensity of the carotid plaque was measured, and the contrast ratio (CR) relative to the adjacent muscle was calculated. RESULTS: CRs of the carotid plaques were 1.39 +/- 0.39, 1.29 +/- 0.29, and 1.23 +/- 0.24 with TRs of 500, 700, and 900 ms, respectively, and were significantly different. Among the plaques, those with a hyperintensity signal (CR > 1.5) and moderate-intensity signal (CR 1.2-1.5) at 500 ms showed a TR dependent signal decrease (hyperintensity plaques, 1.82 +/- 0.26; 1.61 +/- 0.19; and 1.48 +/- 0.17; moderate-intensity plaques, 1.33 +/- 0.08; 1.26 +/- 0.08; and 1.19 +/- 0.07), while those with an isointensity signal (CR < 1.2) remained unchanged regardless of TR (0.96 +/- 0.12, 0.96 +/- 0.11, and 0.97 +/- 0.13). CONCLUSION: The signal intensity of the carotid plaque on T1-weighted imaging significantly varies among different TRs and tends to decrease with longer TR. MR plaque imaging with short and constant TR settings that the ECG-gating method cannot realize would be preferable for evaluating plaque characteristics. PMID- 20020116 TI - Does grazing pressure modify diuron toxicity in a biofilm community? AB - Herbicides affect the structure and functional parameters of fluvial biofilm. Diuron is toxic to primary producers and disrupts endocrine activity. Here, we studied the interaction between this toxicant and several biological compartments in a simple food chain composed of herbivores (the snail Physella [Costatella] acuta) and biofilm. We used indoor experimental channels to which Diuron was added at a realistic concentration (2 mug/L). Bacterial survival and chlorophyll a and photosynthetic activity were analyzed in the biofilm. We monitored biomass, mortality, reproduction, and motility as end points in the freshwater snail P. acuta. Our results showed that bacterial survival and photosynthetic activity were sensitive to Diuron. Snails were not affected by the herbicide at the concentration tested. No significant interactions between the toxicant and grazers were observed on the biofilm. Reproductive traits, however, were slightly affected, indicating a possible endocrine disruption. PMID- 20020117 TI - Acute necrotizing encephalopathy in a child with H1N1 influenza infection. AB - Since the World Health Organization declared a global pandemic of novel influenza A H1N1 in June 2009, there has been a sustained rise in the number of cases of this strain of influenza. Although most cases are mild with complete and uneventful recovery, multiple cases of severe infection with complications including death have been reported. To the best of our knowledge, the majority of fatal outcomes in the United States have been related to pulmonary complications. We report a 12-year-old girl infected with influenza A H1N1 whose clinical course was complicated by rapid progressive neurologic deterioration and striking CT and MRI findings consistent with acute necrotizing encephalopathy (ANE). To our knowledge this has not been reported in the pediatric radiology literature. We hope this case will alert radiologists to this complication and familiarize radiologists with imaging findings that herald ANE. PMID- 20020118 TI - Pediatric eosinophilic esophagitis: radiologic findings with pathologic correlation. AB - BACKGROUND: Eosinophilic esophagitis is increasingly recognized as a cause of dysphagia or food impaction in pediatric patients. It has a high male predominance and is often associated with a history of allergy or asthma. OBJECTIVE: To correlate fluoroscopic findings in eosinophilic esophagitis with the endoscopic and histologic findings. MATERIALS AND METHODS: We retrospectively reviewed the upper gastrointestinal (UGI) findings of eosinophilic esophagitis and correlated them with the clinical, endoscopic and histologic findings in a series of 17 children (12 boys, 5 girls). RESULTS: UGI findings were normal in 12 children, including 4 who had a normal UGI exam after endoscopic disimpaction for an obstructing food bolus. Five children had strictures identified on UGI: one was demonstrated with endoscopy. This suggests that the impactions and strictures were due to an esophageal dysmotility rather than a fixed anatomic abnormality. CONCLUSION: Because the UGI findings are frequently normal in eosinophilic esophagitis, radiologists need to have a high index of suspicion for this disease. In children with a strong clinical history, especially impaction in the absence of an esophageal stricture, endoscopy and biopsy are indicated for further evaluation. PMID- 20020119 TI - Clinical image. Megaloencephalic leukoencephalopathy with subcortical cyst formation (van der Knaap disease). PMID- 20020120 TI - Fetal MR imaging of Kniest dysplasia. AB - We present a case of Kniest dysplasia, a rare form of the type II collagenopathies, with prenatal MRI. Sonography revealed only short limbs in the fetus. Fetal MRI findings included enlarged hyaline cartilaginous structures with abnormally high T2 signal intensity, delayed ossification of the pubic and ischial bones, and platyspondyly. By delineating the cartilaginous abnormalities, fetal MRI can contribute to the prenatal diagnosis of chondrodysplasias. PMID- 20020121 TI - CMEIAS color segmentation: an improved computing technology to process color images for quantitative microbial ecology studies at single-cell resolution. AB - Quantitative microscopy and digital image analysis are underutilized in microbial ecology largely because of the laborious task to segment foreground object pixels from background, especially in complex color micrographs of environmental samples. In this paper, we describe an improved computing technology developed to alleviate this limitation. The system's uniqueness is its ability to edit digital images accurately when presented with the difficult yet commonplace challenge of removing background pixels whose three-dimensional color space overlaps the range that defines foreground objects. Image segmentation is accomplished by utilizing algorithms that address color and spatial relationships of user-selected foreground object pixels. Performance of the color segmentation algorithm evaluated on 26 complex micrographs at single pixel resolution had an overall pixel classification accuracy of 99+%. Several applications illustrate how this improved computing technology can successfully resolve numerous challenges of complex color segmentation in order to produce images from which quantitative information can be accurately extracted, thereby gain new perspectives on the in situ ecology of microorganisms. Examples include improvements in the quantitative analysis of (1) microbial abundance and phylotype diversity of single cells classified by their discriminating color within heterogeneous communities, (2) cell viability, (3) spatial relationships and intensity of bacterial gene expression involved in cellular communication between individual cells within rhizoplane biofilms, and (4) biofilm ecophysiology based on ribotype differentiated radioactive substrate utilization. The stand-alone executable file plus user manual and tutorial images for this color segmentation computing application are freely available at http://cme.msu.edu/cmeias/ . This improved computing technology opens new opportunities of imaging applications where discriminating colors really matter most, thereby strengthening quantitative microscopy-based approaches to advance microbial ecology in situ at individual single-cell resolution. PMID- 20020122 TI - Orientation and dynamics of transmembrane peptides: the power of simple models. AB - In this review we discuss recent insights obtained from well-characterized model systems into the factors that determine the orientation and tilt angles of transmembrane peptides in lipid bilayers. We will compare tilt angles of synthetic peptides with those of natural peptides and proteins, and we will discuss how tilt can be modulated by hydrophobic mismatch between the thickness of the bilayer and the length of the membrane spanning part of the peptide or protein. In particular, we will focus on results obtained on tryptophan-flanked model peptides (WALP peptides) as a case study to illustrate possible consequences of hydrophobic mismatch in molecular detail and to highlight the importance of peptide dynamics for the experimental determination of tilt angles. We will conclude with discussing some future prospects and challenges concerning the use of simple peptide/lipid model systems as a tool to understand membrane structure and function. PMID- 20020123 TI - Effective adoptive therapy of tap-deficient lymphoma using diverse high avidity alloreactive T cells. AB - High avidity for antigen and diversity of T cell receptor (TCR) repertoire are essential for effective immunity against cancer. We have previously created a transgenic mouse strain with increased TCR avidity in a diverse T cell population. In this report, we show that strong alloreactive responses of transgenic T cells against targets with low MHC class I expression can be used for effective adoptive transfer of tumor immunity in vivo. Alloreactive transgenic T cells could be an effective therapeutic approach counteracting tumor evasion of the immune system. PMID- 20020124 TI - Standardization of intraoperative neuromonitoring of recurrent laryngeal nerve in thyroid operation. AB - BACKGROUND: The lack of standardized procedures of intraoperative neuromonitoring (IONM) during thyroid operations may lead to highly variable results, and many of these results can cause misleading information and, conversely, increase the risk of recurrent laryngeal nerve (RLN) injury. Therefore, standardization of IONM procedures is necessary. METHODS: A total of 289 patients (435 nerves at risk) who underwent thyroidectomy by the same surgeon were enrolled in this study. Each patient was intubated with EMG endotracheal tube by the same anesthesiologist. Standardized IONM procedures were applied in each patient. The procedures include preoperative and postoperative video-recording of vocal cord movement, ensuring the correct position of electrodes after the neck was placed at full extension, vagal stimulation and registration of EMG signals before and after RLN dissection, and photographic documentation of the exposed RLN. RESULTS: Five patients encountered dysfunction of IONM, which was caused by malposition of electrodes and the problem was detected at once. One patient with non-RLN was detected at the earlier stage of operation. Eighteen nerves experienced loss of EMG signals during thyroid dissection, and the causes of nerve injuries were well elucidated with the application of our standardized IONM procedures. CONCLUSIONS: The standardized IONM procedures are useful and helpful not only to eliminate false IONM results, but also to elucidate the mechanism of RLN injury. After ascertaining the surgical pitfalls and improving the surgical techniques, the palsy rate was significantly reduced in this study. PMID- 20020125 TI - Teres minor innervation in the context of isolated muscle atrophy. AB - PURPOSE: Teres minor atrophy occurs either in isolation, associated with other rotator cuff muscle pathologies or in quadrilateral space syndrome. In the latter condition, compression of the axillary nerve is the likely cause; however, the anatomy of the nerve to teres minor and how this may relate to isolated teres minor atrophy have not been extensively investigated. In light of the significance of teres minor atrophy in shoulder pathology, we performed a combined radiological and anatomical study of teres minor and its nerve supply. METHODS: Cadaveric dissection of nine shoulder specimens from eight cadavers was performed to investigate the anatomical variability in course, length and branching pattern of both the teres minor nerve and the axillary nerve. Radiological imaging and reports were analysed on all shoulder magnetic resonance images performed over a 1-week period at four radiology clinic locations in an attempt to identify the incidence of isolated teres minor atrophy and review teres minor atrophy in association with other shoulder pathology. Finally, we studied a case of isolated teres minor atrophy identified during a routine undergraduate dissection class. RESULTS: Considerable anatomical variation was noticed in cadaver dissections in the nerve(s) supplying teres minor muscle revealing several various points where it may be vulnerable to impingement or injury at along its course. Analysis of 61 shoulder MR images revealed two patients with shoulder complaints that had isolated teres minor atrophy. Case based study of these two male patients revealed other associated shoulder injury but the presentation was markedly different and clinically distinct from quadrilateral space syndrome. CONCLUSION: Isolated teres minor atrophy is a relatively common shoulder pathology which appears to be clinically distinct from other syndromes with rotator cuff muscle atrophy including quadrilateral space syndrome. The exact aetiology is unknown but cadaveric dissection in this study suggests the considerable anatomical variation in both the origin and length of teres minor nerve(s) increase the risk of impingement and subsequent isolated teres minor atrophy. PMID- 20020126 TI - CTLA-4 +49A>G polymorphism of recipients of HLA-matched sibling allogeneic stem cell transplantation is associated with survival and relapse incidence. AB - Conflicting observations have been reported about the role of CTLA-4 gene polymorphisms in the clinical outcome of allogeneic hematopoietic stem cell transplantation (HSCT). We have investigated three polymorphisms of the CTLA-4 gene (-318C>T, +49A>G, CT60G>A) in 133 donor/recipient pairs who underwent HLA matched sibling donor HSCT for hematological malignancies. We found no association of the clinical outcome of the HSCT with either recipient or donor 318C>T and CT60G>A polymorphisms. At variance, we found a significant association of donor +49A>G G/G genotype with longer overall survival (OS; log-rank test, P = 0.04), and the number of +49A>G G-alleles in the recipient with longer OS (P = 0.027), longer disease-free survival (P = 0.036) and reduced relapse rate (P = 0.042). However, only recipient +49A>G polymorphism was retained as independent prognostic factor in a multivariate analysis, suggesting that the expression of CTLA-4 on the cells of recipient may be relevant for the clinical outcome of HSCT. PMID- 20020127 TI - The role of chemokines in B cell chronic lymphocytic leukaemia: pathophysiological aspects and clinical impact. AB - Chemokines are centrally involved in leukocyte migration, homing and haematopoiesis. Besides these physiological aspects, their role in pathological processes especially with respect to solid tumour and haematological neoplasias is well established. In this context, the focus was set here on disclosing their contribution in B cell chronic lymphocytic leukaemia (B-CLL), which is regarded as the most characteristic low-grade lymphoma. Up to now, it has been demonstrated that several chemokines are involved in migration of B-CLL cells to lymph nodes, secondary lymphoid organs and bone marrow. Moreover, some chemokines are known to have an anti-apoptotic effect and thus contribute to the survival of B-CLL cells. By interfering with both of these aspects, new therapeutic targets for this yet incurable disease may be developed. Furthermore, a correlation can be drawn between the concentration of some chemokines in patients' serum, the expression of their respective receptors on B-CLL cells and well-established predictive clinical parameters. Consequently, further systematic investigation of the chemokine network may lead to the identification of new diagnostic and prognostic markers. This review focuses on the impact of chemokines and their receptors on B-CLL pathophysiology and points out potential implications for both treatment and diagnosis. PMID- 20020128 TI - Novel microtubule-targeting agents, pyrrolo-1,5-benzoxazepines, induce apoptosis in multi-drug-resistant cancer cells. AB - PURPOSE: The development of multi-drug resistance (MDR) due to the expression of members of the ATP binding cassette (ABC) transporter family is a major obstacle in cancer treatment. The broad range of substrate specificities associated with these transporters leads to the efflux of many anti-cancer drugs from tumour cells. Therefore, the development of new chemotherapeutic agents that are not substrates of these transporters is important. We have recently demonstrated that some members of a novel series of pyrrolo-1,5-benzoxazepine (PBOX) compounds are microtubule-depolymerising agents that potently induce apoptosis in several cancer cell lines and impair growth of mouse breast tumours. The aim of this current study was to establish whether PBOXs were capable of inducing apoptosis in cancer cells expressing either P-glycoprotein or breast cancer resistance protein (BCRP), two of the main ABC transporters associated with MDR. METHODS: We performed in vitro studies to assess the effects of PBOXs on cell proliferation, cell cycle and apoptosis in human cancer cell lines and their drug-resistant substrains expressing either P-glycoprotein or BCRP. In addition, we performed a preliminary molecular docking study to examine interactions between PBOXs and P glycoprotein. RESULTS: We established that three representative PBOXs, PBOX-6, 15 and -16 were capable of inducing apoptosis in drug-resistant HL60-MDR1 cells (expressing P-glycoprotein) and HL60-ABCG2 cells (expressing BCRP) with similar potencies as in parental human promyelocytic leukaemia HL60 cells. Likewise, resistance to PBOX-6 and -16 was not evident in P-glycoprotein-expressing A2780 ADR cells in comparison with parent human ovarian carcinoma A2780 cells. Finally, we deduced by molecular docking that PBOX-6 is not likely to form favourable interactions with the substrate binding site of P-glycoprotein. CONCLUSION: Our results suggest that pro-apoptotic PBOX compounds may be potential candidates for the treatment of P-glycoprotein- or BCRP-associated MDR cancers. PMID- 20020130 TI - A Lagrangian particle method for reaction-diffusion systems on deforming surfaces. AB - Reaction-diffusion processes on complex deforming surfaces are fundamental to a number of biological processes ranging from embryonic development to cancer tumor growth and angiogenesis. The simulation of these processes using continuum reaction-diffusion models requires computational methods capable of accurately tracking the geometric deformations and discretizing on them the governing equations. We employ a Lagrangian level-set formulation to capture the deformation of the geometry and use an embedding formulation and an adaptive particle method to discretize both the level-set equations and the corresponding reaction-diffusion. We validate the proposed method and discuss its advantages and drawbacks through simulations of reaction-diffusion equations on complex and deforming geometries. PMID- 20020129 TI - Cellular and molecular mechanisms for the synergistic cytotoxicity elicited by oxaliplatin and pemetrexed in colon cancer cell lines. AB - PURPOSE: Oxaliplatin effect in the treatment of colorectal cancer is improved upon combination with thymidylate synthase (TS) inhibitors. Pemetrexed is polyglutamated by the folylpolyglutamate synthase (FPGS) and blocks folate metabolism and DNA synthesis by inhibiting TS, dihydrofolate reductase (DHFR) and glycinamide ribonucleotide formyltransferase (GARFT). The present study evaluates the pharmacological interaction between oxaliplatin and pemetrexed in colorectal cancer cells. METHODS: Human HT29, WiDr, SW620 and LS174T cells were treated with oxaliplatin and pemetrexed. Drug interaction was studied using the combination index method, while cell cycle was investigated with flow cytometry. The effects of drugs on Akt phosphorylation and apoptosis were studied with ELISA and fluorescence microscopy, respectively. RT-PCR analysis was performed to assess whether drugs modulated the expression of pemetrexed targets and of genes involved in DNA repair (ERCC1 and ERCC2). Finally, platinum-DNA adduct levels were detected by ultra-sensitive multi-collector inductively coupled plasma mass spectrometry (ICP-MS). RESULTS: A dose-dependent inhibition of cell growth was observed after drug exposure, while a synergistic interaction was detected preferentially with sequential combinations. Oxaliplatin enhanced cellular population in the S-phase. Drug combinations increased apoptotic indices with respect to single agents, and both drugs inhibited Akt phosphorylation. RT-PCR analysis showed a correlation between the FPGS/(TS x DHFR x GARFT) ratio and pemetrexed sensitivity, as well as a downregulation of ERCC1, ERCC2, TS, DHFR and GARFT after drug exposure. In addition, pretreatment with pemetrexed resulted in an increase of oxaliplatin-DNA adducts. CONCLUSION: These data demonstrate that oxaliplatin and pemetrexed synergistically interact against colon cancer cells, through modulation of cell cycle, inhibition of Akt phosphorylation, induction of apoptosis and modulation of gene expression. PMID- 20020131 TI - Etanercept and anakinra can prolong febrile episodes in patients with hyperimmunoglobulin D and periodic fever syndrome. AB - Hyperimmunoglobulin D and periodic fever syndrome (HIDS) is a rare, hereditary autoinflammatory condition, characterized by recurrent inflammatory episodes. There is no proven treatment for HIDS, but various drugs including, non-steroidal anti-inflammatory drugs, colchicine, steroids, statins and thalidomide have all been tried. Recently, some patients have demonstrated a good clinical response to either etanercept or anakinra. We report a case of a 10-year-old girl who experienced prolonged and severe inflammatory attacks, when she was treated with etanercept, and later with anakinra. PMID- 20020132 TI - Treatment of relapsing polychondritis in the era of biological agents. AB - Relapsing polychondritis (RP) is a rare disorder, often requiring high doses of immunosuppressive therapy to control its potentially life-threatening consequences. The advent of biological agents has added to the armamentarium available to treat RP, but the lack of controlled trials, along with the small numbers of patients and disease heterogeneity means that new therapies are prescribed without the benefits of rigorous clinical research. Thus, information on individual cases is of value in expanding our knowledge of the use of biologic agents in rare conditions. We report on the use of rituximab in a patient who subsequently developed catastrophic aortic incompetence, and we review the literature in relation to the use of this drug in RP. PMID- 20020133 TI - Validation of the World Health Organization disability assessment schedule II (WHODAS-II) in patients with osteoarthritis. AB - The objective of this study is to test the reliability and validity of WHODAS-II (Turkish version) for the assessment of disability in patients with osteoarthritis. This study is designed as follows: the internal construct validity of WHODAS-II in patients with knee osteoarthritis was assessed by Rasch analysis, and external construct validity by association with the Western Ontario and McMaster Universities Index of Osteoarthritis (WOMAC) and the Nottingham Health Profile (NHP); reliability was tested by internal consistency, intra-class correlation coefficient (ICC) and test-retest ICC. Our study presents the results of 225 outpatients assessed with mean age 58.4 years (SD 11.1) of whom 80.9% were female. Cronbach's alpha, ICC, and test-retest ICC values for the six subscales of WHODAS-II varied between 0.71 and 0.94, 0.71 and 0.94, and 0.87 and 0.97, respectively. Rasch analysis of WHODAS-II indicated that after adjustment for local dependency, satisfactory fit was achieved. Two separate 'activities' and 'participation' components could also be identified. External construct validity of the scale was confirmed with expected correlations with WOMAC and NHP. This study concludes that WHODAS-II provides a reliable and valid health status instrument for measuring disability and components of 'activities' and 'participation' in patients with osteoarthritis. Thus, it provides the opportunity to model the consequences of disease according to the International Classification of Functioning, Disability and Health framework. PMID- 20020134 TI - A sequence of pathologic events in a patient after thymectomy for myasthenia gravis. AB - The case of a rare coexistence of myasthenia gravis (MG) with systemic lupus erythematosus (SLE) is described. MG was diagnosed prior to SLE which developed after thymectomy. The patient was affected by HCV viremia. Therefore, there were therapeutic problems. Metylase treatment was continued for several years and dopamine receptor agonist was effectively administered as adjunctive therapy in SLE. PMID- 20020135 TI - Cementless total hip arthroplasty in patients with rheumatoid arthritis using a tapered designed titanium hip stem minimum: 10-year results. AB - The results of cementless tapered designed femoral stem were studied at a minimum 10-year follow-up in a non-selected, consecutive group of 27 patients (39 hips) with rheumatoid arthritis. Clinical and radiological analyses were performed in 27 hips, 17 patients (mean age at surgery 45 years) after a mean of 12 years. The postoperative Harris hip score was excellent for 14 hips, 9 hips were rated as good and 4 hips were fair or poor. No stem had to be revised for aseptic loosening. Proximal stress shielding was observed in 26 hips (96%); heterotopic ossification was present in 11 hips (41%). Six hips required revision of the acetabular component. With uncemented tapered femoral fixation excellent 12-year results are achieved in patients with rheumatoid arthritis. PMID- 20020136 TI - A retrospective review of rheumatology referral wait times within a health centre in Quebec, Canada. AB - It is important that inflammatory arthropathies such as rheumatoid arthritis be diagnosed promptly so that treatment can be administered in a timely fashion. However, there is considerable evidence that this process of care is delayed in many people. The aim of the study is to assess wait times between primary care referral and rheumatology assessment for new-onset inflammatory arthropathies. We performed a retrospective review related to new rheumatology consultations (N = 202) between September and November 2008 within the McGill University Health Centre, Montreal, Canada. At this centre, no formal triaging of rheumatology referrals exists. Of the 202 charts reviewed, wait times could be calculated in 164 cases. Only consultations for new-onset conditions were analyzed (N = 161). The results showed that patients with inflammatory arthritis were seen approximately 34.6 days (median 26) post-referral. Wait times for individuals who were ultimately diagnosed with non-urgent conditions (osteoarthritis, fibromyalgia and soft-tissue rheumatism) averaged 41.0 days (median 29). In conclusions, compared to non-urgent cases, individuals with inflammatory arthritis were seen about 1 week sooner. Nevertheless, provisional diagnosis provided on referrals did not appear to expedite wait times for persons with suspected inflammatory arthritis. This suggested that other factors, such as the concern of the patient, may have an influence on referral wait times. Implementation of a rapid access program or triage system may be helpful to further decrease wait times for inflammatory arthropathies. PMID- 20020137 TI - Serum amyloid A is a useful marker to evaluate the disease activity of Takayasu's arteritis. AB - Takayasu's arteritis (TA) is a rare large vessel vasculitis that is difficult to diagnose in the early stages. Therefore, it is also very difficult to manage and prevent irreversible vascular damage in TA. A 19-year-old female patient with back pain was examined using [(18)F]-FDG-PET to detect the source of inflammation. Specific accumulation of [(18)F]-FDG was observed in the thoracic and abdominal aorta, leading to the diagnosis of TA. Corticosteroid treatment resulted in clinical remission. However, the serum amyloid A (SAA) levels remained elevated. A follow-up scan showed residual uptake of [(18)F]-FDG in the thoracic aorta suggesting subclinical vascular inflammation. Methotrexate was combined with the corticosteroid, and the elevated levels of SAA became normalized. The present case suggests that monitoring serum levels of SAA and [(18)F]-FDG-PET could help clinicians to make adequate treatment adjustments in TA patients. PMID- 20020139 TI - A case of cellulitis causing tissue defect during etanercept therapy. AB - Tumor necrosis factor-alpha (TNF-alpha) antagonists are employed increasingly during recent years in patients with active rheumatoid arthritis who do not respond to disease-modifying anti-rheumatic drugs. Contraindications such as infections, auto-antibody formation and hypersensitive reactions can be observed during the treatment with TNF-alpha antagonist drugs. Our case was a 52-year-old woman, followed by several centers for a period of 21 years with a seropositive, erosive and nodular RA diagnosis. Anti TNF-alpha treatment was commenced due to the failure to control the disease. During the treatment, a serious cellulite developed, which required hospitalization and surgical debridement as well as intravenous antibiotics treatment. Through the present case, we aimed to draw attention to the skin infection during the use of etanercept in a patient with RA. PMID- 20020138 TI - Adult-onset Still's disease. AB - Adult-onset Still's disease (AOSD) is a rare systemic inflammatory disorder characterised by high spiking fever, an evanescent salmon pink rash and arthritis, frequently accompanied by sore throat, myalgias, lymphadenopathies, splenomegaly and neutrophilic leukocytosis. Aetiology is still unknown, however, it seems that an important role is played by various infectious agents, which would act as triggers in genetically predisposed hosts. Diagnosis is a clinical one and may be lengthy because it requires exclusion of infectious neoplasms, including malignant lymphomas and leukaemias, and other autoimmune diseases. Different diagnostic or classification criteria have been proposed, but not definitely accepted. There are no specific laboratory tests for AOSD, but they reflect the systemic inflammation: the ESR is consistently high, while the rheumatoid factors and antinuclear antibodies are negative. High serum ferritin levels associated with a low fraction of its glycosylated component are assessed as useful diagnostic and disease activity markers. The clinical course can be divided into three main patterns with different prognoses: self-limited or monophasic, intermittent or polycyclic systemic and chronic articular pattern. Therapy includes non-steroidal anti-inflammatory drugs, corticosteroids and disease modifying anti-rheumatic drugs: biological agents have recently been introduced and they seem to be very promising not only for the treatment but also for understanding the pathogenic mechanisms underlying the disease. PMID- 20020140 TI - Willingness to pay for improvement of physical function among rheumatoid arthritis patients as measured by Health Assessment Questionnaire. AB - The main objective of this study was to assess the symptoms and functional difficulties caused by rheumatoid arthritis through application of the willingness to pay (WTP) method. Structured questionnaire study was conducted among 242 RA patients. The subjects were asked to evaluate their functional capacity using visual analog scales (VAS) for all the 20 questions in the Health Assessment Questionnaire (HAQ). Each VAS was followed by a question asking how much the respondent would be willing to pay on a monthly basis for a 50% improvement with the function in question. These were combined with later collected data on clinical status and use of RA-related health services. The average WTP varied greatly in the examined 20 different functions. The total WTP average on a 50% improved functional capacity amounted to 567.05 per month. Patients with lower functional capacity (HAQ 1.2 or more) were ready to contribute significantly (p < 0.001) more (705 euros/month) than those with better functional status (199 euros/month). Subjects, whose financial standing was better, reported a higher total WTP (r = 0.218, p < 0.01). On average, the total WTP of the respondents equaled 18.44% of the disposable net monthly income per person of the household. However, the variation was quite substantial. WTP among patients with rheumatoid arthritis was best depicted through the functional capacity of the patient, the possible time the patient had retired, and the global feeling of pain. In conclusion, monetary value is a concrete way of portraying subjective valuation. WTP method is suitable for assessing functional deficits of rheumatoid arthritis. PMID- 20020141 TI - Which is the dominant factor for perception of rheumatic pain: meteorology or psychology? AB - It is believed that there is an association between the weather and rheumatic symptoms. We aimed to investigate what kind of association is present and what are the factors which determine the nature of this association. Fifty-six subjects with rheumatic disease (31 RA, 15 SpA, 10 OA) who live in Antalya were followed between December 2005 and July 2006. Patients were asked to fill diaries which contain questions regarding the symptoms of their rheumatic diseases everyday. In every monthly visit, disease activity measurement, laboratory assessment and Beck depression inventory assessment were recorded. The symptomatic and psychological measurements were matched with the meteorological data of Antalya Regional Directorate of Meteorological Service of Turkish State. Correlation of symptoms with weather variables was investigated. Contributory effect of weather and of psychologic factors on symptom scores were evaluated by stepwise multiple regression analysis. Eighty-four percent of subjects belive in an association between weather and rheumatism, while 57% claimed to have ability to forecast weather. The maximum correlation coefficient between weather and arthritis symptoms was -0.451 and the maximum contribution of weather on symptoms was 17.1%. Arthritis symptoms were significantly contributed by Beck depression score. The belief about presence of weather-arthritis association was found to be stronger than its statistical power. Our results did not prove or rule out the presence of weather-rheumatism association. As long as the scientific attempts result in failure, the intuitive support in favour of the presence of weather arthritis association will go on forever. PMID- 20020142 TI - Joint-specific prevalence and radiographic pattern of hand osteoarthritis in Korean. AB - We investigated the prevalence and involvement patterns of radiographic osteoarthritis (OA) with hand symptoms among Korean people and compared the difference in prevalence of hand OA between racial groups. Hand radiographs in 299 Korean subjects (266 female, 33 male) >=40 years of age were examined, who had hand arthralgia. The study population was comprised of 206 patients who had radiographic OA at least at one hand joint. Radiographic OA (Kellgren-Lawrence scale >=2 grades) was evaluated for 16 joints of each hand. The most prevalent OA was in the interphalangeal joints (IP) of thumb, followed by the distal interphalangeal joints (DIP) of index finger, DIP of middle and fifth finger in the frequency of order. The involvement of metacarpophalangeal joints (MP) was relatively common in 1st-3rd MP. Hand OA in Korean was higher in the thumb IP and lower in the thumb carpometacarpal joints compared with Caucasian previously reported. Moreover, the higher OA frequency of 1st-3rd MP was not in accordance with other studies in Caucasian and other Asian populations. The patterns of radiographic hand OA were symmetric (OR 15.68), clustered by ray (OR 8.69) and row (OR 6.66). In conclusion, our study showed that thumb IP and 2nd/3rd/5th DIP should be included in the assessment of radiologic hand OA in Koreans. PMID- 20020143 TI - Prevalence of and risk factors for low bone mineral density in Japanese female patients with systemic lupus erythematosus. AB - To examine the prevalence of and risk factors for low bone mineral density (BMD) (osteoporosis or osteopenia) in Japanese female patients with systemic lupus erythematosus (SLE). We performed BMD measurements by dual X-ray absorptiometry at the lumbar spine and the hip and collected basic and lifestyle-related, clinical and treatment characteristics among 58 SLE patients. Odds ratios (ORs) and their 95% confidence intervals (CIs) were assessed for associations between low BMD and selected factors among SLE patients. The mean BMD +/- SD was 0.90 +/- 0.17 g/cm(2) at the lumbar spine and 0.76 +/- 0.17 g/cm(2) at the hip. The prevalence of osteopenia (2.5 SD < T score < 1 SD) was 50.0% and that of osteoporosis (T score < 2.5 SD) was 13.8% in our SLE patients. After adjustment for age and disease duration, we found the number of deliveries (OR = 5.58, 95% CI = 1.31-26.06; P = 0.02) to be a risk factor for overall low BMD (T score < 1 SD) and a maximal dosage of >50 mg/day of oral corticosteroids (OR = 0.25, 95% CI = 0.07-0.91; P = 0.035) as a preventive factor for low BMD at the lumbar spine. Reduced BMD, especially in spinal trabecular bone, was pronounced in Japanese female patients with SLE, particular in those with a history of delivery. A history of high-dose oral corticosteroids was associated with the preservation of BMD at the lumbar spine, however, further study is needed considering the limited sample size. PMID- 20020144 TI - A field model of learning: 2. Long-term memory in the crab Chasmagnathus granulatus. AB - In the previous companion paper, the possibility of learning by Chasmagnathus in field conditions was demonstrated. Here, we study long-term memory inquiring to what extent an internal representation could be maintained in a complex environment. Two 45-min training sessions, each of 15 visual danger stimulus presentations with 3-min intertrials, were given at a 24-h interval. Throughout the first training session and during the first 22.5 min of re-training on day 2, crabs kept the same hiding response level but then, during the second phase of re training, the re-emerging increased up to the point that 32% of surface crabs ignored the stimulus. Each session was followed by a 22.5-min testing without stimulation. At testing on day 2 after re-training, crabs showed a change in the usual exploring strategy. Results reveal long persistency in responding despite a rest interval of 24 h followed by a gradual decrease in response until it vanishes. The statistical analysis was performed by comparing performances at day 2 (Rescorla in Am Psychol 43:151-160, 1988) and then confirmed through comparisons between day 1 and day 2. However, it is not possible to identify separate and taxonomically well-defined learnings but rather a tangled collection of processes that influence each other blurring some of the diagnostic features of each learning. PMID- 20020145 TI - A field model of learning: 1. Short-term memory in the crab Chasmagnathus granulatus. AB - Learning and memory studies have been performed for more than two decades using the crab Chasmagnathus in our laboratory. Here, our research was aimed at disclosing some instances of learning in field conditions. Three experiments were performed non-simultaneously, all with a 22.5-min pre-training preceding the first visual-danger-stimulus, an opaque rectangle passing overhead. In Experiment 1, crabs received a single stimulus followed by 22.5-min testing without stimulation, where the re-emerging latency was considered the basic latency response. In Experiment 2, training consisted of 15 stimulus 3-min apart, followed by 22.5-min testing without stimulation. Throughout training crabs were underground but re-emerged at testing with latencies longer than the basic latency response. Both at pre-training and testing the usual strategy of exploring was the short-near excursions. In Experiment 3, training included three stimulus 22.5-min apart, followed by 22.5-min testing. Crabs left their burrows before the end of each inter-trial, showing a mean latency like the basic latency response, but a sensitization to the stimulus and a preponderance of the fast-far excursions over the usual slow-near. In brief: through 15-3 training, crabs learn that the stimulus is iteratively presented; through 3-22.5 training, crabs acquire sensitization to the stimulus and a different strategy of exploration. PMID- 20020147 TI - A modified Delorme's operation for the treatment of rectal mucosal prolapse. AB - AIMS: Many procedures have been described for the treatment of rectal internal mucosal prolapse (RMP), but the therapeutic effect is questionable. This work is to evaluate clinical and functional outcome of a modified Delorme's operation trans-anal purse-string sutures for rectal mucosa and submucosa (TAS) for the treatment of RMP. METHODS: The clinical data of 58 patients with rectal mucosal prolapse treated with rectal mucosa and submucosal tissue (TAS) between June 2004 and June 2008 were analyzed retrospectively. RESULTS: No patient died. Satisfaction with surgery was high in 48 cases (82.8%), moderate in seven (12.1%), and low in three (5.17%). Prolapse relapse rate was 5.17%. Anal tenesmus and urge to defecate resolved in 52 (89.7%) patients. Constipation improved in 25 of 28 (89.3%) previously constipated patients. No surgery-associated constipation occurred. The mean operative time was 31 (range 22-46) min. Mean hospital stay was 3 days (range 2-6). Mean patient follow-up was 32 months (range 12-60). CONCLUSIONS: From our data, TAS for the treatment of RMP showed encouraging results with little complications and an acceptable relapse rate. This economical procedure induces only mild trauma and is easy to perform, making it worthy of further practice and investigation. PMID- 20020146 TI - Immunohistological evidence for Wnt-signaling activation in Peutz-Jeghers polyposis. AB - OBJECTIVE: Molecular pathogenesis of gastrointestinal polyposis in Peutz-Jegher's syndrome (PJS) has been linked to the loss-of-function mutation of LKB1. Recent functional genetic studies have pointed out that LKB1 plays a physiological role in controlling the Wnt-signaling pathway and activation of the pathway as a consequence of LKB1 haploinsufficiency might be responsible for the development of harmatomatous polyps. This study aimed to look for immunohistochemical evidence of Wnt-signaling activation in PJS polyps. METHOD: Beta-catenin immunohistochemistry patterns were evaluated in gastrointestinal polyps from five cases of PJS. All patients were also evaluated for germline mutations of LKB1 and somatic mutations of beta-catenin in the polyps. RESULTS: Four of the five cases had germline mutations of LKB1, including two novel mutations, a one-base insertion at codon 53 and a large deletion encompassing exon 3 (codon 136-155). PJS polyps from all patients showed generalized membrane and cytoplasmic localizations of beta-catenin along the mucosal endothelium. Polyps from two cases with LKB1 mutations revealed moderate-intensity nuclear staining in approximately 20 and 70% of the polyps. CONCLUSION: The study offers additional evidence of Wnt-signaling activation in PJS polyp development at the tissue level, although the degree of up-regulation was not as high as has been found in Wnt-associated neoplasms. PMID- 20020148 TI - Giant cell tumor of the femoral head and neck: result of intralesional curettage. AB - BACKGROUND: There have been not much reports on the result of intralesional excision for giant cell tumors (GCTs) of the femoral head and neck because of its rarity. The purpose of this study is to review the results of patients managed with intralesional curettage for GCT of the femoral head and neck. METHODS: We retrospectively reviewed 12 patients with a GCT of the femoral head and/or neck. All of them were treated with curettage and followed up to monitor local recurrence and pulmonary metastasis. Mean duration of follow-up was 58.3 months. RESULTS: Although recurrence rate of the present study was rather high (41.7%, 5 of 12 hips), 9 of 12 hips (75%) were preserved at last follow-up including 2 hips that underwent repeat curettage, and functional outcomes of the preserved hips were satisfactory. CONCLUSION: It may be that curettage should be considered as a primary treatment of choice for GCTs of the femoral head and neck. PMID- 20020149 TI - Cervical spondylolysis in a judo player: a case report and biomechanical analysis. PMID- 20020150 TI - Tubercular endometritis visualized as endometrial micropolyps during hysteroscopic procedure. AB - Female genital tuberculosis is a rare disease in developed countries but it is a frequent cause of chronic pelvic inflammatory disease and infertility in undeveloped countries. A rare case of tubercular endometritis diagnosed at hysteroscopy and successfully treated in a woman with secondary infertility, is presented. As far as we know this is the first case that shows the association between endometrial micropolyps and tubercular endometritis. PMID- 20020151 TI - Takayasu's arteritis in pregnancy complicated by peripartum aortic dissection. AB - INTRODUCTION: We describe a patient with a known diagnosis of Takayasu's arteritis who presented late in the third trimester. She was delivered by caesarean section and her postpartum course was complicated by aortic dissection. METHOD: This report is the first to describe peripartum aortic dissection in a patient with known Takayasu's arteritis. CONCLUSION: Takayasu's arteritis should be regarded as a risk factor for aortic dissection. It is important to treat systemic hypertension in Takayasu's arteritis patients and suspect the diagnosis of aortic dissection in any pregnant patient complaining of chest pain as dissection is a leading cause of maternal mortality in the developed world. PMID- 20020152 TI - Clastogenic plasma factors: a short overview. AB - A large number of studies have revealed that irradiated subjects produce soluble factors found in their blood plasma which, when transferred into cell cultures from non-irradiated individuals, show clastogenic (chromosome breaking) activity. Increased yields of chromatid-type aberrations have been characteristic in most of these studies. Exposed cohorts of various origins have revealed to possess this feature: from A-bomb survivors to patients treated with radiotherapy. It is apparent that the plasma factors are sustainable for long time periods. On the other hand, they seem to be produced very fast after exposure. Considerable variation in the effect has been found between individuals with similar radiation exposure. Further, the phenomenon is not restricted to irradiated populations. Clastogenic plasma has also been observed in patients with inflammatory diseases or congenital chromosome breakage syndromes as well in subjects exposed to other agents than ionizing radiation. Chromosomal aberration inducing substances have been detected not only in vivo, but also in vitro. A common feature to all the conditions is that they are associated with oxidative stress. Studies on the biochemical nature of the clastogenic factor(s) have been conducted, and tumor necrosis factor alpha and lipid peroxidation products, among others, have been suggested as good candidates. The relevance of the plasma factors to health effects remains open. The aim of the paper is to give a short overview on the phenomenon of clastogenic factors--their occurrence and formation as well as possible effectors. PMID- 20020153 TI - Cystic tumour of the atrioventricular node: three cases of sudden death. AB - Cystic tumour of the atrioventricular nodal region is a rare primary cardiac tumour that can cause sudden death. Antemortem diagnosis and successful excision of this type of tumour are extremely rare. Three cases of sudden death are reported: A 25- and 40-year-old with a history of heart block and a 60-year-old with no medical history. There have been more than 120 cases of sudden death attributed to primary cardiac tumours in the literature. Although over 100 of these lesions were histologically benign, their intracardiac locations precipitated conductive and haemodynamic abnormalities that resulted in sudden death. The most common intracardiac lesion causing sudden death, cystic tumour of the atrioventricular node, however, may not be discovered unless the atrioventricular node is microscopically examined. PMID- 20020155 TI - Action plans produce separate Simon effects for picking up and transporting objects. AB - A common type of interaction with the environment is reaching to move an object from one location to another. We investigated the influence of spatial stimulus response (S-R) compatibility effects on the planning of such multi-reach actions. Participants responded to a cue on their left or right side by picking up a bean (initial reach) and then placing it in a container (goal reach). In three experiments, the initial reach and the goal reach were varied so that both reaches were to opposite sides (Exp 1), the initial reach was neutral (Exp 2), and both the initial and goal reaches were on the same side (Exp 3). The participants' reaction time to begin the initial reach was measured. Results showed that the spatial compatibility between the cue and both the initial and goal reaches contribute to the reaction time, with the relation between the cue and the initial reach playing a dominant role. PMID- 20020156 TI - Clinical practice: management of biliary atresia. AB - Biliary atresia is a rare, serious and challenging disease in newborn children. Its aetiology remains unknown. Optimal management at specialist centres with resultant better overall outcomes is achieved through a multidisciplinary team approach. The Kasai portoenterostomy performed early in life remains the only surgical repair procedure. Two thirds of patients will clear their jaundice after a Kasai procedure, but only about one third will retain their livers after the first decade of life. Failure of this procedure leaves liver transplantation as the only chance for survival, and this disease is the commonest indication for liver transplantation in children. With modern medical care and refinements in surgical techniques, survival after either or both of these procedures is about 90%. Early referral to specialist centres and long-term specialist care remains the key to successful treatment of this condition. PMID- 20020157 TI - Spatial and temporal variability of plant-available water in calcium carbonate cemented soils and consequences for arid ecosystem resilience. AB - Increased variability in precipitation, including frequency of drought, is predicted for many arid and semiarid regions globally. The ability of soils to retain water can increase resilience by buffering vegetation communities against precipitation extremes. Little is known, however, about water retention by carbonate-cemented soil horizons, which occur extensively in arid and semiarid ecosystems. It has been speculated that they may significantly modify vertical and temporal distribution of plant-available water (PAW). To investigate this hypothesis, PAW was monitored at three sites in a mixed shrub-grass community in southern New Mexico, USA, across soils with differing degrees of carbonate horizon development: no carbonate horizon, a horizon partially cemented with carbonates (calcic), and a horizon continuously cemented with carbonates (petrocalcic). Results are presented from 3 years that included extremely dry and wet periods. Both carbonate-cemented horizons absorbed and retained significantly greater amounts of PAW for several months following an extremely wet winter and summer compared to the non-carbonate soil. Following a wet summer, continuously cemented horizons retained very high PAW (16-18% volumetric or approximately 72 80% of soil water holding capacity) through early spring of the following year, more than double the PAW retained by similar depths in the non-carbonate soil. Drying dynamics indicate both carbonate-cemented horizons release stored water into the grass rooting zone during growing seasons following extreme wet events. Water dynamics of these horizons during extreme events provide a mechanism to explain previous observations that perennial grasses exhibit greater resilience to drought when carbonate-cemented horizons occur at shallow depths (<50 cm). Water holding capacity of the entire profile, including horizons cemented with carbonates, should be considered when evaluating the potential resilience of vegetation communities to disturbance, including the increased variability in precipitation expected to occur as a result of global climate change. PMID- 20020158 TI - Changes in glomerular mesangium in kidneys with congenital nephrotic syndrome of the Finnish type. AB - Congenital nephrotic syndrome of the Finnish type (NPHS1, CNF) is an autosomal recessive disease caused by mutations in a major podocyte protein, nephrin. NPHS1 is associated with heavy proteinuria and the development of glomerular scarring. We studied the cellular and molecular changes affecting the glomerular mesangium in NPHS1 kidneys. Marked hyperplasia of mesangial cells (MC) was mainly responsible for the early mesangial expansion in NPHS1 glomeruli. The levels of the proliferation marker, mindbomb homolog 1 and the major MC mitogen, platelet derived growth factor, and its receptors, however, were quite normal. Only a small number of cells were positive for CD68 (marker for phagocytic cells) and CD34 (marker for mesenchymal precursor cells) in the NPHS1 mesangium. MCs strongly expressed alpha-smooth muscle actin, indicating myofibloblast transformation. The expression levels of the profibrotic mediators osteopontin and transforming growth factor beta were up-regulated in NPHS1 glomeruli by 3.2 and 1.6-fold, respectively, compared to the controls. The synthesis by MCs of the typical fibroblast products collagen I, fibronectin, and tenascin, however, was low, and the extracellular matrix increase was caused by the accumulation of a normal MC product, collagen IV. The results indicate that severe glomerular sclerosis can develop without major qualitative cellular or molecular changes in the mesangium. PMID- 20020159 TI - Multidetector computed tomography findings and correlations with proteinuria in nutcracker syndrome. AB - We evaluated the effectiveness of multidetector computed tomography (MDCT) as a diagnostic tool for nutcracker syndrome (NS) and its association with proteinuria. The angle and distance between the aorta and the superior mesenteric artery (SMA), the degree of difference in corticomedullary enhancement (DCE) between kidneys in the nephrographic phase of computed tomography, peak velocity ratio (PVR), and anteroposterior diameter ratio (APDR) in the sonogram were measured. The MDCT results, sonogram results, and the ratio of protein:creatinine were significantly different between NS patients and the controls. The area under the curve for angle, distance, and DCE were 0.895 +/- 0.058, 0.876 +/- 0.063, and 0.942 +/- 0.036, respectively. The cutoff values for angle and distance had sensitivity and specificity values of 96.2 and 80% for <22.4 degrees and 84.6 and 80% for <4.9 mm, respectively. The DCE had a sensitivity of 88.5% and a specificity of 100% for the positive scores. There were significant correlations between the degree of DCE and the ratio of protein:creatinine (r = 0.337, p = 0.031), and between distance and the ratio of protein:creatinine (r = -0.419, p = 0.006). We conclude that MDCT has diagnostic value for NS in children and that MDCT findings are correlated with proteinuria. PMID- 20020161 TI - Luminal sulfide and large intestine mucosa: friend or foe? AB - Hydrogen sulfide (H(2)S) is present in the lumen of the human large intestine at millimolar concentrations. However, the concentration of free (unbound) sulfide is in the micromolar range due to a large capacity of fecal components to bind the sulfide. H(2)S can be produced by the intestinal microbiota from alimentary and endogenous sulfur-containing compounds including amino acids. At excessive concentration, H(2)S is known to severely inhibit cytochrome c oxidase, the terminal oxidase of the mitochondrial electron transport chain, and thus mitochondrial oxygen (O(2)) consumption. However, the concept that sulfide is simply a metabolic troublemaker toward colonic epithelial cells has been challenged by the discovery that micromolar concentration of H(2)S is able to increase the cell respiration and to energize mitochondria allowing these cells to detoxify and to recover energy from luminal sulfide. The main product of H(2)S metabolism by the colonic mucosa is thiosulfate. The enzymatic activities involved in sulfide oxidation by the colonic epithelial cells appear to be sulfide quinone oxidoreductase considered as the first and rate-limiting step followed presumably by the action of sulfur dioxygenase and rhodanese. From clinical studies with human volunteers and experimental works with rodents, it appears that H(2)S can exert mostly pro- but also anti-inflammatory effects on the colonic mucosa. From the available data, it is tempting to propose that imbalance between the luminal concentration of free sulfide and the capacity of colonic epithelial cells to metabolize this compound will result in an impairment of the colonic epithelial cell O(2) consumption with consequences on the process of mucosal inflammation. In addition, endogenously produced sulfide is emerging as a prosecretory neuromodulator and as a relaxant agent toward the intestinal contractibility. Lastly, sulfide has been recently described as an agent involved in nociception in the large intestine although, depending on the experimental design, both pro- and anti-nociceptive effects have been reported. PMID- 20020160 TI - Cutting-edge information for the management of acute pancreatitis. AB - Considering that the Japanese (JPN) guidelines for the management of acute pancreatitis were published in Takada et al. (J HepatoBiliary Pancreat Surg 13:2 6, 2006), doubts will be cast as to the reason for publishing a revised edition of the Guidelines for the management of acute pancreatitis: the JPN guidelines 2010, at this time. The rationale for this is that new criteria for the severity assessment of acute pancreatitis were made public on the basis of a summary of activities and reports of shared studies that were conducted in 2008. The new severity classification is entirely different from that adopted in the 2006 guidelines. A drastic revision was made in the new criteria. For example, about half of the cases that have been assessed previously as being 'severe' are assessed as being 'mild' in the new criteria. The JPN guidelines 2010 are published so that consistency between the criteria for severity assessment in the first edition and the new criteria will be maintained. In the new criteria, severity assessment can be made only by calculating the 9 scored prognostic factors. Severity assessment according to the contrast-enhanced computed tomography (CT) grade was made by scoring the poorly visualized pancreatic area in addition to determining the degree of extrapancreatic progress of inflammation and its extent. Changes made in accordance with the new criteria are seen in various parts of the guidelines. In the present revised edition, post-endoscopic retrograde cholangiopancreatography (ERCP) pancreatitis is treated as an independent item. Furthermore, clinical indicators (pancreatitis bundles) are presented to improve the quality of the management of acute pancreatitis and to increase adherence to new guidelines. PMID- 20020162 TI - Amino acids special issue 'Protein interactions in the virus-host relationship'. PMID- 20020163 TI - Compatible and self-incompatible pollination in Pyrus communis displays different polyamine levels and transglutaminase activity. AB - The polyamine (PA) content and the transglutaminase (TGase) activity have been investigated in Pyrus communis pollination with compatible and self-incompatible (SI) pollen in order to deepen their possible involvement in the progamic phase of plant reproduction. The PA distribution as free, perchloric acid (PCA)-soluble and PCA-insoluble fractions in ungerminated (UGP), germinating pollen (GP), styles and pollinated styles with compatible and SI pollens is discussed in the light of a possible role during pollination. Generally, the conjugated PAs both in PCA-soluble and PCA-insoluble fractions were higher than the free form. Within the conjugated PAs, the PCA-insoluble ones were the highest with the exception of the not pollinated styles. As TGase mediates some of the effects of PAs by covalently binding them to proteins, the activity of this enzyme, never checked before in styles and pollinated styles, was examined. In the SI styles, the TGase activity is higher in comparison to style-pollinated with compatible pollen, and high molecular mass cross-linked products were formed, suggesting an involvement of TGase in SI response. This is the first evidence on the presence of this enzyme activity in not pollinated and pollinated styles. PMID- 20020164 TI - Adapting and testing telephone-based depression care management intervention for adolescent mothers. AB - This phase 1 clinical trial combined qualitative and quantitative methods to modify a collaborative care, telephone-based, depression care management intervention for adolescent mothers and to determine the acceptability, feasibility, and initial efficacy of the intervention in a sample of adolescent mothers (n = 97) who were recruited from a Teen Parent Program. Outcomes included measures of depressive symptoms, functioning, and use of mental health services. Acceptability of the intervention was demonstrated, but feasibility issues related to the complex life challenges confronting the adolescent mother. Although only four adolescent mothers received mental health treatment, there was a trend for improved depressive symptoms over time. Results of the study provide data for the need of further refinement of the intervention before a large clinical trial is conducted for adolescent mothers with symptoms of depression. PMID- 20020165 TI - The C15orf2 gene in the Prader-Willi syndrome region is subject to genomic imprinting and positive selection. AB - C15orf2 (Chromosome 15 open reading frame 2) is an intronless gene, which is located in the Prader-Willi syndrome (PWS) chromosomal region on human chromosome 15. Mice do not have an orthologous gene. Here we show that expression of C15orf2 in the fetal human brain is imprinted. Using Western blot and immunohistological studies we have obtained evidence that C15orf2 protein is present in several regions of the brain. Previously published phylogenetic studies as well as population genetic studies based on complex haplotypes as described here suggest that C15orf2 is under positive Darwinian selection. These results indicate that C15orf2 might have an important role in human biology and that a deficiency of C15orf2 might contribute to PWS. PMID- 20020166 TI - The performance of rooks in a cooperative task depends on their temperament. AB - In recent years, an increasing number of studies demonstrated the existence of consistent individual differences in behaviour, often referred to as differences in temperament or personality, in a wide range of animal species. There notably is a growing body of evidence showing that individuals differ in their propensity for risk taking or reacting to stressful situations. This variation has been related to differences in learning abilities or performance in cognitive tasks. In the present study, we examined the consequences of inter-individual variation in boldness on performance in a cooperative task in rooks (Corvus frugilegus). Birds were tested individually to measure a number of behavioural parameters related to boldness. The level of a stress-related hormone, corticosterone, in the faeces of each bird was measured under control conditions and after a stress provoking event. In parallel, we conducted a cooperative string pulling task in which birds were tested in dyads. Successful cooperation depended to a large extent on the temperament of the two partners involved. Temperament, in turn, correlated well with corticosterone levels under stress. Bolder individuals appeared to be more willing to participate in the task, whereas shyer individuals were more influenced by the behaviour of their partner. These findings suggest that a rook's temperament can limit its options of forming successfully cooperating partnerships under stressful conditions. PMID- 20020167 TI - What matters in the associative learning of visual cues in foraging parasitoid wasps: colour or brightness? AB - Visual cues are known to be used by numerous animal taxa to gather information on quality and localisation of resources. Because environmental lighting can interfere with the spectral features of visual cues, the specific characteristics of the colour signals that promote forager decision and learning are still not known in the majority of insects (excepted in bees). We analysed the effect of previous experience on the use of visual information by the wasp Venturia canescens, a parasitoid of pyralidae, in the context of host searching. These parasitoids search for hosts concealed in several fruit species, so visual cues from the host microhabitat could play a key role in host finding. We also investigated the type of visual cues on which wasps based their decision. We tested whether wasps are able to associate an achromatic cue (brightness) or a chromatic one (hue, i.e. dominant wavelength and/or chroma) with the presence of hosts. Our results show that in the context of host foraging, chromatic cues are more reliable than brightness in achieving the associative learning process. Therefore, understanding the behavioural ecology of foraging should make use of the knowledge about the visual information used. PMID- 20020168 TI - Discrimination of human and dog faces and inversion responses in domestic dogs (Canis familiaris). AB - Although domestic dogs can respond to many facial cues displayed by other dogs and humans, it remains unclear whether they can differentiate individual dogs or humans based on facial cues alone and, if so, whether they would demonstrate the face inversion effect, a behavioural hallmark commonly used in primates to differentiate face processing from object processing. In this study, we first established the applicability of the visual paired comparison (VPC or preferential looking) procedure for dogs using a simple object discrimination task with 2D pictures. The animals demonstrated a clear looking preference for novel objects when simultaneously presented with prior-exposed familiar objects. We then adopted this VPC procedure to assess their face discrimination and inversion responses. Dogs showed a deviation from random behaviour, indicating discrimination capability when inspecting upright dog faces, human faces and object images; but the pattern of viewing preference was dependent upon image category. They directed longer viewing time at novel (vs. familiar) human faces and objects, but not at dog faces, instead, a longer viewing time at familiar (vs. novel) dog faces was observed. No significant looking preference was detected for inverted images regardless of image category. Our results indicate that domestic dogs can use facial cues alone to differentiate individual dogs and humans and that they exhibit a non-specific inversion response. In addition, the discrimination response by dogs of human and dog faces appears to differ with the type of face involved. PMID- 20020169 TI - A new method for studying problem solving and tool use in stingrays (Potamotrygon castexi). AB - Testing the cognitive abilities of cartilaginous fishes is important in understanding the evolutionary origins of cognitive functions in higher vertebrates. We used five South American fresh water stingrays (Potamotrygon castexi) in a learning and problem-solving task. A tube test apparatus was developed to provide a simple but sophisticated procedure for testing cognitive abilities of aquatic animals. All five subjects quickly learned to use water as a tool to extract food from the testing apparatus. The experimental protocol, which gave the animals the opportunity of correcting a wrong visual cue decision, resulted in four out of five subjects correcting an error rather than making an initial right choice. One of five subjects reached 100% correct trials in the visual discrimination task. The ability to use water as an agent to extract food from the testing apparatus is a first indication of tool use in batoid fishes. Performance in the instrumental task of retrieving food from a novel testing apparatus and the rapid learning in the subsequent discrimination/error correction task shows that cartilaginous fish can be used to study the origins of cognitive functions in the vertebrate lineage. PMID- 20020170 TI - A review of current European treatment guidelines for migraine. AB - Migraine is one of the ten most disabling disorders worldwide, and despite recent developments in the management of migraine, it remains underdiagnosed and undertreated. Guidelines for the management of migraine aim to improve the quality of patient care and to assist professionals in decision making in relation to the overall healthcare process. Most European countries have published national clinical practice guidelines for migraine treatment. These guidelines need to be kept up-to-date with the most recent best clinical evidence and therapeutic strategies to ensure their optimal use to improve health outcomes. The aim of this review is to compare the English language guidelines available across Europe, analyzing differences and similarities, in order to provide a general overview to assist in assessing whether a European consensus on migraine treatment can be achieved. PMID- 20020171 TI - FaRE1: a transcriptionally active Ty1-copia retrotransposon in strawberry. AB - Retrotransposons are ubiquitous in the plant kingdom and constitute a large fraction of many plant genomes. Although most retrotransposons from plants were thought to be transcriptionally silent in somatic tissues, evidence of activity under certain conditions is available in some cases. In this study, a complete LTR retrotransposon was isolated from the cultivated strawberry (Fragaria x ananassa) genome using genome walking. The element, named FaRE1, has all the features of a typical Ty1-copia retrotransposon. Its total length was 5,104 bp, comprising a single 3,891 bp open reading frame. It is represented by approximately 96 copies per genome, equivalent to approximately 0.33% of the genome. Transcription of FaRE1 was detected in leaf tissue treated with various phytohormones, such as naphthalene acetic acid, 2,4-dichlorophenoxyacetic acid or abscisic acid . To our knowledge, this is the first report of the isolation of a complete LTR retrotransposon with transcriptional activity in strawberry. PMID- 20020172 TI - Chloroplast DNA phylogeography of Betula maximowicziana, a long-lived pioneer tree species and noble hardwood in Japan. AB - Betula maximowicziana is an ecologically and economically important tree species in Japan. In order to examine the phylogeographical pattern of the species in detail, maternally inherited chloroplast (cp) DNA variations of 25 natural populations of Betula maximowicziana and a total of 12 populations of three related species were evaluated by PCR-RFLP analysis. Two main haplotypic groups of B. maximowicziana populations (northern and southern) were detected, with the main boundary passing through the Tohoku region in northeastern Japan; in addition there was high genetic differentiation among the 25 populations studied (GST = 0.950, G'ST =0:977). The phylogeographical pattern exhibited by B. maximowicziana was much more similar to that of alpine plants than to that of beech and oak. Comparison of the patterns of genetic structure obtained from the cpDNA with previously and newly acquired data on bi-parentally inherited nuclear DNA indicates that the nuclear genome was transferred via pollen from the northern haplotypic group to the southern group more frequently than it moved in the opposite direction. Although common haplotypes were detected among B. maximowicziana and the two related species examined, these haplotypes were not shared sympatrically, suggesting very rare hybridization among the species currently occurring in their natural populations. PMID- 20020173 TI - Effects of seasonal and interannual variations in leaf photosynthesis and canopy leaf area index on gross primary production of a cool-temperate deciduous broadleaf forest in Takayama, Japan. AB - Revealing the seasonal and interannual variations in forest canopy photosynthesis is a critical issue in understanding the ecological mechanisms underlying the dynamics of carbon dioxide exchange between the atmosphere and deciduous forests. This study examined the effects of temporal variations of canopy leaf area index (LAI) and leaf photosynthetic capacity [the maximum velocity of carboxylation (V (cmax))] on gross primary production (GPP) of a cool-temperate deciduous broadleaf forest for 5 years in Takayama AsiaFlux site, central Japan. We made two estimations to examine the effects of canopy properties on GPP; one is to incorporate the in situ observation of V (cmax) and LAI throughout the growing season, and another considers seasonality of LAI but constantly high V (cmax). The simulations indicated that variation in V (cmax) and LAI, especially in the leaf expansion period, had remarkable effects on GPP, and if V (cmax) was assumed constant GPP will be overestimated by 15%. Monthly examination of air temperature, radiation, LAI and GPP suggested that spring temperature could affect canopy phenology, and also that GPP in summer was determined mainly by incoming radiation. However, the consequences among these factors responsible for interannual changes of GPP are not straightforward since leaf expansion and senescence patterns and summer meteorological conditions influence GPP independently. This simulation based on in situ ecophysiological research suggests the importance of intensive consideration and understanding of the phenology of leaf photosynthetic capacity and LAI to analyze and predict carbon fixation in forest ecosystems. PMID- 20020174 TI - The use of chronic disease risk factor surveillance systems for evidence-based decision-making: physical activity and nutrition as examples. AB - OBJECTIVE: To highlight the value of continuous risk factor surveillance systems in providing evidence of the impact of, and to inform health promotion interventions. METHOD: An ongoing risk factor surveillance system involving telephone interviews with approximately n = 600 randomly selected South Australians each month. Trend analysis on physical activity (PA) levels and daily consumption of fruit and vegetables was undertaken. RESULTS: An apparent seasonal trend for fruit consumption and PA was found, with less activity and fruit consumption undertaken in winter months. Overweight/obese adults exercised less than those with normal BMI, and females less than males, although PA rates for both females and overweight/obese adults are rising. There was an increase in vegetable consumption following a major media campaign. Although reported prevalence of the consumption of five or more serves of vegetables daily and the mean number of serves consumed daily has decreased, it is still above pre campaign rates. CONCLUSION: Additional information obtained from a risk factor surveillance system, when compared to an annual or point-in-time survey, provides valuable evidence for health professionals interested in measuring and assessing the effectiveness of health promotion interventions. PMID- 20020175 TI - German wide cross sectional survey on health impacts of electromagnetic fields in the view of general practitioners. AB - OBJECTIVES: The proportion of general practitioners (GPs) in Germany who assume health impacts of electromagnetic fields (EMF) is assessed. Moreover, factors associated with this risk perception are examined. METHODS: A 7% random sample was drawn from online lists of all the GPs working in Germany. 1,867 doctors received a long version of a self-administered postal questionnaire about EMF and health (response rate 23.3%), 928 doctors received a short version (response rate 49.1%). RESULTS: 37.3% of responders to the short and 57.5% of responders to the long questionnaire agreed "that there are persons whose health complaints are caused by EMF when legal limit values are met". A late responder analysis for the survey with the short questionnaire led to a still lower estimate of 29% for GPs believing in health-relevant effects of EMF. CONCLUSION: About a third of German GPs associate EMF with health complaints and thus deviate considerably from current scientific knowledge. To avoid a strong selection bias in the surveys of the perception of EMF risks, use of short questionnaires and late responder analysis are recommended. PMID- 20020176 TI - Perceived factors related to cigarette and waterpipe (ghelyan) initiation and maintenance in university students of Iran. AB - OBJECTIVES: To investigate the perceived factors leading to cigarette and waterpipe (ghelyan) smoking initiation and maintenance in Iranian university students. METHOD: A cross-sectional study was conducted on 233 students aged 20 25 enrolled in Isfahan University and Kashan University in 2007. Demographic factors as well as cigarette and ghelyan status, and perceived factors related to cigarette or ghelyan smoking initiation and maintenance were recorded. Principal component analysis was carried out to cluster perceived smoking initiation and maintenance factors. RESULT: In a multiple logistic regression model, perceived psychological factors were more significant to cigarette smoking initiation and maintenance than ghelyan smoking among girls (odds ratio, OR = 1.90; P < 0.04) although perceived psychological factors (OR = 2.20; P < 0.001) and social factors (OR = 2.42; P < 0.001) were more significant in cigarette smoking initiation than ghelyan smoking initiation among boys (OR = 2.42; P < 0.001). Moreover, study appears that in boys, perceived entertainment factors could play more significant role in ghelyan smoking initiation compared with cigarette smoking initiation (OR = 0.42; P < 0.001). CONCLUSION: Whereas the factors related to smoking initiation and maintenance are different in both genders and in both tobacco products, identification of cigarette and ghelyan smoking may guide policy makers to develop comprehensive interventions. PMID- 20020178 TI - Primary diffuse cerebral leptomeningeal atypical teratoid rhabdoid tumor: report of the first case. AB - Atypical teratoid rhabdoid tumor (AT/RT) of the central nervous system has been recently described as a distinct clinicopathological entity with characteristic morphologic, immunophenotypic and molecular characteristics. AT/RT typically involves the posterior fossa of the pediatric population. Supratentorial AT/RT is exceedingly rare. In this report, we describe a very unusual case of a child who presented with signs and symptoms suggestive of leptomeningitis. However, imaging studies and histologic findings showed plaque-like AT/RT involving the leptomeninges of the cerebrum, cerebellum, and spinal cord. The disease proved to be rapidly fatal and resulted in the patient's death within approximately two weeks. To our knowledge, this is the first case of primary leptomeningeal AT/RT involving the supratentorial leptomeninges. PMID- 20020177 TI - Construction of a ganciclovir-sensitive lentiviral vector to assess the influence of angiopoietin-3 and soluble Tie2 on glioma growth. AB - Malignant brain tumors grow by coopting the existing vasculature, a process involving the release of angiopoietin-2 (Angpt2) from endothelial cells and its binding to the Tie2 receptor. The first goal of this study was to examine the therapeutic potential of two proteins that could interfere with Angpt2, namely Angpt3 and the soluble extracellular domain of Tie2 (sTie2). The second goal was to develop a lentiviral vector capable of delivering such proteins while offering the possibility to identify and destroy the genetically modified cells. To this end, we designed a bicistronic construct expressing the marker enhanced green fluorescent protein fused to the suicide gene herpes simplex virus 1 thymidine kinase. GL261 glioma cells transduced with this vector could be tracked and killed on command by the administration of the prodrug ganciclovir, either in vitro or after implantation into mouse brains. High levels of Angpt3 or sTie2 could be achieved with this vector; however, Angpt3 increased capillary destabilization and glioma growth, whereas sTie2 exerted no effect. Overall, this study helps to understand the importance of the Tie2 signaling pathway in glioma development and the role of Angpt3, but suggests that neither this molecule nor sTie2 are effective agents against malignant gliomas. This study also provides a lentiviral vector design for safer gene therapy. PMID- 20020179 TI - Bevacizumab fails to treat temporal paraganglioma: discussion and case illustration. AB - Temporal paragangliomas are highly vascular tumors treated primarily by surgical resection. However, surgery to remove these tumors is associated with significant morbidity, including cranial nerve dysfunction. Interestingly, these tumors have been shown to express vascular endothelial growth factor (VEGF). A variety of tumors expressing VEGF and the VEGF receptor have been shown to reduce in size and vascularity when treated with the VEGF-specific antibody, bevacizumab (Avastin). We hypothesized that paragangliomas may be treated noninvasively with bevacizumab, either as a primary treatment or as a useful adjuvant to surgical resection or radiation. Thus, our aim was to evaluate the effects of bevacizumab on this patient's paraganglioma. A 36-year-old female presented to us with a 3 month history of positional dizziness, light-headedness, and left ear pulsatile tinnitus and hearing loss. She was found to have a temporal paraganglioma (glomus jugulare tumor) on imaging. Histopathology confirmed significant staining for VEGF. This patient was treated with bevacizumab prior to surgical treatment; radiographic imaging at 3 months, however, showed no significant response. We discuss possible reasons for treatment failure. PMID- 20020180 TI - Monotherapy with methotrexate for primary central nervous lymphoma has single agent activity in the absence of radiotherapy: a single institution cohort. AB - We have retrospectively reviewed toxicities and response of a cohort of primary central nervous system lymphoma (PCNSL) patients treated with high dose parenteral methotrexate (MTX) monotherapy without whole brain radiation. From The Massachusetts General Hospital (MGH) Cancer Registry, active since 1946, we selected all immunocompetent patients with histologic and/or radiographic PCNSL diagnosed between 1980 and 2007. We identified the recipients of MTX with leucovorin rescue as sole therapy. No patient received radiation therapy (XRT). We analyzed this cohort for toxicity, response and patterns of recurrence. The cohort of 121 patients received on average 11 cycles of intravenous MTX at a median dose of 8 g/m(2). Median interval between cycles was 10 days. After 3 months of therapy, the overall response rate was 85% (58% CR, 27% PR). The overall survival (OS) for the cohort was 7 years and progression-free survival (PFS) was 3.14 years. A trend toward a higher PFS was seen in patients who continued to receive MTX (3.48 years) every three months as compared to patients who ceased MTX after one year (2.86 years). Of 68 patients who achieved initial CR, there were 40 recurrences. Twenty-six of the 40 were re-induced with MTX as above; Sixty-nine percent again achieved CR. Eighty-one treatment-related toxicities occurred in 1316 MTX cycles. These toxicities included MRI white matter changes (N = 8) and lead to MTX cessation in 16 patients. High-dose MTX monotherapy of PCNSL is well-tolerated and provides PFS of >3 years and OS >7 years. PMID- 20020181 TI - Artificial neural network study on organ-targeting peptides. AB - We report a new approach to studying organ targeting of peptides on the basis of peptide sequence information. The positive control data sets consist of organ targeting peptide sequences identified by the peroral phage-display technique for four organs, and the negative control data are prepared from random sequences. The capacity of our models to make appropriate predictions is validated by statistical indicators including sensitivity, specificity, enrichment curve, and the area under the receiver operating characteristic (ROC) curve (the ROC score). VHSE descriptor produces statistically significant training models and the models with simple neural network architectures show slightly greater predictive power than those with complex ones. The training and test set statistics indicate that our models could discriminate between organ-targeting and random sequences. We anticipate that our models will be applicable to the selection of organ-targeting peptides for generating peptide drugs or peptidomimetics. PMID- 20020182 TI - Acute and/or chronic stress models modulate CuZnSOD and MnSOD protein expression in rat liver. AB - Cellular protection against oxidative stress is afforded by the enzyme superoxide dismutase (SOD). In this study, the protein levels of copper-zinc SOD (CuZnSOD) in the cytosolic and nuclear fraction, manganese SOD (MnSOD) in the mitochondrial, and cytosolic fraction and cytochrome c (cyt c) in the liver of male rats exposed to 2 h of acute immobilization (IM) or Cold stress, 21 days chronic isolation or their combinations (chronic/acute stress) were examined. The serum corticosterone (CORT) level was measured, as an indicator of stress stimuli. Both acute stressors with elevated CORT levels caused a decrease of mitochondrial MnSOD, while acute IM resulted in redistribution of the CuZnSOD protein level between the cytosolic and nuclear fraction. Chronic isolation, during which the CORT level was close to control value, resulted in an increase of cytosolic CuZnSOD, whereas a decrease of MnSOD in mitochondrial and its corresponding increase in cytosol fraction was found. In both combined stress regimes, an increase of the CuZnSOD and MnSOD levels in the cytosolic fraction was recorded whereby increase of the CORT level was observed only in the chronic isolation followed by acute IM. The data indicate that acute and/or chronic stress models have different degrees of influence on serum CORT and SOD subcellular protein levels. Increased cytosolic CuZnSOD protein level under chronic isolation suggests that state of oxidative stress may also exist under CORT level similar to the basal value. The presence of MnSOD and cyt c in the cytosolic fraction could serve as useful parameters for mitochondrial dysfunction. PMID- 20020183 TI - Casein kinase 2 inhibition differentially modulates apoptotic effect of trichostatin A against epithelial ovarian carcinoma cell lines. AB - Histone deacetylase inhibitors and casein kinase 2 inhibitors have been shown to induce apoptosis. However, the combined effect of casein kinase 2 inhibition on the apoptotic effect of histone deacetylase inhibitor is unknown. We assessed the effect of casein kinase 2 inhibition on the apoptotic effect of trichostatin A in human epithelial carcinoma cell lines with respect to cell death signaling pathways. At concentrations that did not induce cell death, the casein kinase 2 inhibitor 4,5,6,7-tetrabromobenzotriazole inhibited activation of apoptotic proteins and changes in mitochondrial membrane permeability induced by the histone deacetylase inhibitor trichostatin A. These results suggest that casein kinase 2 inhibition may reduce trichostatin A-induced apoptosis in ovarian carcinoma cell lines by suppressing activation of apoptotic proteins and changes in mitochondrial membrane permeability, which both lead to caspase-3 activation. Casein kinase 2 inhibition, which does not induce a cytotoxic effect, may prevent histone deacetylase inhibitor-mediated apoptosis. PMID- 20020184 TI - Sequence and expression of 11beta-hydroxysteroid dehydrogenase type 1 cDNA cloned from pig testis. AB - Pig 11beta-hydroxysteroid dehydrogenase (11beta-HSD) type 1 cDNA was cloned from neonatal pig testis, and 15 nucleotides were found to differ from the sequence in GenBank (Accession No. NM_214248). It was an exclusive clone obtained as pig 11beta-HSD type 1, and the sequence of 11beta-HSD type 1 cDNA cloned from pig liver was identical to that from testis. Amino acid sequence, deduced from cloned cDNA, also had a conserved triad of catalytically important Ser, Tyr and Lys residues for the short-chain dehydrogenase/reductase family, a membrane-spanning domain consisting of hydrophobic amino acid and a glycine motif in the cofactor binding region. The protein translated from this clone on expression in mammalian HEK293 cells exhibited oxo-reduction activity of cortisone and oxidation activity of cortisol. Furthermore, this oxo-reduction activity of cortisone was stimulated by co-expression of human H6PDH, while oxidation activity of cortisol was suppressed by H6PDH co-expression in HEK293 cells. Based on these results, the sequence of newly cloned cDNA is considered to correspond to an active enzyme form of pig 11beta-HSD type 1. PMID- 20020185 TI - Monitoring cellular behaviour using Raman spectroscopy for tissue engineering and regenerative medicine applications. AB - Raman spectroscopy has been used to determine the chemical composition of materials for over 70 years. Recent spectacular advances in laser and CCD camera technology creating instruments with higher sensitivity and lower cost have initiated a strong resurgence in the technique, ranging from fundamental research to process control methodology. One such area of increased potential is in tissue engineering and regenerative medicine (TERM), where autologous cell culture, stem cell biology and growth of human cells on biomaterial scaffolds are of high importance. Traditional techniques for the in vitro analysis of biochemical cell processes involves cell techniques such as fixation, lysis or the use of radioactive or chemical labels which are time consuming and can involve the perpetuation of artefacts. Several studies have already shown the potential of Raman spectroscopy to provide useful information on key biochemical markers within cells, however, many of these studies have utilised micro- or confocal Raman to do this, which are not suited to the rapid and non-invasive monitoring of cells. For this study a versatile fit-for-purpose Raman spectrometer was used, employing a macro-sampling optical platform (laser spot size 100 mum at focus on the sample) to discriminate between different TERM relevant cell types and viable and non-viable cells. The results clearly show that the technique is capable of obtaining Raman spectra from live cells in a non-destructive, rapid and non invasive manner, however, in these experiments it was not possible to discriminate between different cell lines. Despite this, notable differences were observed in the spectra obtained from viable and non-viable cells, showing significant changes in the spectral profiles of protein, DNA/RNA and lipid cell constituents after cell death. It is evident that the method employed here shows significant potential for further utilisation in TERM, providing data directly from live cells that fits within a quality assurance framework and provides the opportunity to analyse cells in a non-destructive manner. PMID- 20020186 TI - Bio-corrosion characterization of Mg-Zn-X (X = Ca, Mn, Si) alloys for biomedical applications. AB - The successful applications of magnesium-based alloys as biodegradable orthopedic implants are mainly inhibited due to their high degradation rates in physiological environment. This study examines the bio-corrosion behaviour of Mg 2Zn-0.2X (X = Ca, Mn, Si) alloys in Ringer's physiological solution that simulates bodily fluids, and compares it with that of AZ91 magnesium alloy. Potentiodynamic polarization and electrochemical impedance spectroscopy results showed a better corrosion behaviour of AZ91 alloy with respect to Mg-2Zn-0.2Ca and Mg-2Zn-0.2Si alloys. On the contrary, enhanced corrosion resistance was observed for Mg-2Zn-0.2Mn alloy compared to the AZ91 one: Mg-2Zn-0.2Mn alloy exhibited a four-fold increase in the polarization resistance than AZ91 alloy after 168 h exposure to the Ringer's physiological solution. The improved corrosion behaviour of the Mg-2Zn-0.2Mn alloy with respect to the AZ91 one can be ascribed to enhanced protective properties of the Mg(OH)(2) surface layer. The present study suggests the Mg-2Zn-0.2Mn alloy as a promising candidate for its applications in degradable orthopedic implants, and is worthwhile to further investigate the in vivo corrosion behaviour as well as assessed the mechanical properties of this alloy. PMID- 20020187 TI - Validation of arterial blood pressures observed from the patient monitor; a tool for prehospital research. AB - OBJECTIVES: For some time, the inaccuracies of non-invasive blood pressure measurement in critically ill patients have been recognised. Measurement difficulties can occur even in optimal conditions, but in prehospital transportation vehicles, problems are exacerbated. Intra-arterial pressures must be used as the reference against which to compare the performance of non-invasive methods in the critically ill patient population. Intra-arterial manometer data observed from the patient monitor has frequently been used as the reference against which to assess the accuracy of noninvasive devices in the emergency setting. To test this method's validity, this study aimed to determine whether numerical monitor pressures can be considered interchangeable with independently sampled intra-arterial pressures. METHODS: Intensive Care Unit nurses were asked to document arterial systolic, diastolic and mean pressures numerically displayed on the patient monitor. Observed pressures were compared to reference intra arterial pressures independently recorded to a computer following analogue to digital conversion. Differences between observed and recorded pressures were evaluated using the Association for the Advancement of Medical Instrumentation (AAMI) protocol. Additionally, two-level linear mixed effects analyses and Bland Altman comparisons were undertaken. RESULTS: Systolic, diastolic and integrated mean pressures observed during 60 data collection sessions (n = 600) fulfilled AAMI protocol criteria. Integrated mean pressures were the most robust. For these pressures, mean error (reference minus observed) was 0.5 mm Hg (SD 1.4 mm Hg); 95% CI (two-level linear mixed effects analysis) 0.4-0.6 mm Hg; P < 0.001. Bland Altman plots demonstrated tight 95% limits of agreement (-2.3 to 3.2 mm Hg), and uniform agreement across the range of mean blood pressures. CONCLUSIONS: Integrated mean arterial pressures observed from a well maintained patient monitor can be considered interchangeable with independently sampled intra arterial pressures and may be confidently used as the reference against which to test the accuracy of non-invasive blood pressure measuring methods in the prehospital or emergency setting. PMID- 20020188 TI - The longitudinal consistency of mother-child reporting discrepancies of parental monitoring and their ability to predict child delinquent behaviors two years later. AB - This study examined the longitudinal consistency of mother-child reporting discrepancies of parental monitoring and whether these discrepancies predict children's delinquent behaviors 2 years later. Participants included 335 mother/female-caregiver and child (46% boys, >90% African American; age range 9 16 years [M = 12.11, SD = 1.60]) dyads living in moderate-to-high violence areas. Mother-child discrepancies were internally consistent within multiple assessment points and across measures through a 2-year follow-up assessment. Further, mothers who at baseline consistently reported higher levels of parental monitoring relative to their child had children who reported greater levels of delinquent behaviors 2 years later, relative to mother-child dyads that did not evidence consistent discrepancies. This finding could not be accounted for by baseline levels of the child's delinquency, maternal and child emotional distress, or child demographic characteristics. This finding was not replicated when relying on the individual reports of parental monitoring to predict child delinquency, suggesting that mother-child reporting discrepancies provided information distinct from the absolute frequency of reports. Findings suggest that mother-child discrepancies in reports of parental monitoring can be employed as new individual differences measurements in developmental psychopathology research. PMID- 20020189 TI - Acculturation and sexual risk behaviors among Latina adolescents transitioning to young adulthood. AB - Latinas in the United States are at a disproportionate risk for STDs and sexual risk behaviors. Among Latinas, acculturation has been found to be one of the most important predictors of these behaviors. Therefore, this study examined the longitudinal association between Latina adolescents' level of acculturation and multiple sexual risk outcomes, including self-report STD diagnosis, four or more life-time sex partners, regret of sexual initiation after alcohol use, and lack of condom use during young adulthood. Based on the National Longitudinal Study of Adolescent Health (Add Health), this study includes a nationally representative sample of 1,073 Latina adolescents (ages 11-20 at Wave 1) transitioning into young adulthood (ages 18-27 at Wave 3). Our findings indicate that more acculturated Latinas who spoke English at home were more likely to have STDs and to exhibit sexual risk behaviors than Latinas who were foreign-born and did not use English at home. Interventions that aim to promote sexual and reproductive health among young Latinas should take into consideration their different levels of acculturation. This approach holds greater potential for reducing health disparities among Latinas. PMID- 20020190 TI - The impact of multiple types of child maltreatment on subsequent risk behaviors among women during the transition from adolescence to young adulthood. AB - The purpose of this study was to investigate how different types of child maltreatment, independently and collectively, impact a wide range of risk behaviors that fall into three domains: sexual risk behaviors, delinquency, and suicidality. Cumulative classification and Expanded Hierarchical Type (EHT) classification approaches were used to categorize various types of maltreatment. Data were derived from Wave III of the National Longitudinal Study of Adolescent Health (Add Health). Our sample consisted of White, Black, Hispanic, and Asian females ages 18 to 27 (n = 7,576). Experiencing different kinds of maltreatment during childhood led to an extensive range of risk behaviors within the three identified domains. Women experiencing sexual abuse plus other maltreatment types had the poorest outcomes in all three domains. These findings illustrate that it may no longer be appropriate to assume that all types of maltreatment are equivalent in their potential contribution to negative developmental sequelae. PMID- 20020191 TI - Anti-inflammatory and immunomodulatory effects of Ulomoides dermestoides on induced pleurisy in rats and lymphoproliferation in vitro. AB - The following study aimed to evaluate, in vitro and in vivo, the anti inflammatory effect of Ulomoides dermestoides, a beetle commonly used as a remedy for a variety of diseases including respiratory disorders and asthma. We used an acute inflammation model of injury, injection of carrageenan into the pleural cavity of rats. The rats were treated intraperitoneally with the aqueous extract of U. dermestoides 8 and 16 mg/kg. The exudate volume, protein concentration, polymorphonuclear leukocytes (PMNs) and total leukocyte were measured. The peripheral blood mononuclear cells (PBMCs) were isolated from the blood of healthy subjects and we investigated the immunomodulatory and cytotoxic effect of aqueous extract of U. dermestoides. In conclusion, in vitro we observed a non cytotoxic effect and antiproliferative activity on the dose of 12.5 mg/dL. In vivo, this paper clarifies the great clinical relevance of the aqueous extract of U. dermestoides in elucidating its role as an anti-inflammatory agent. PMID- 20020192 TI - Pseudo-Lyapunov exponents and predictability of Hodgkin-Huxley neuronal network dynamics. AB - We present a numerical analysis of the dynamics of all-to-all coupled Hodgkin Huxley (HH) neuronal networks with Poisson spike inputs. It is important to point out that, since the dynamical vector of the system contains discontinuous variables, we propose a so-called pseudo-Lyapunov exponent adapted from the classical definition using only continuous dynamical variables, and apply it in our numerical investigation. The numerical results of the largest Lyapunov exponent using this new definition are consistent with the dynamical regimes of the network. Three typical dynamical regimes-asynchronous, chaotic and synchronous, are found as the synaptic coupling strength increases from weak to strong. We use the pseudo-Lyapunov exponent and the power spectrum analysis of voltage traces to characterize the types of the network behavior. In the nonchaotic (asynchronous or synchronous) dynamical regimes, i.e., the weak or strong coupling limits, the pseudo-Lyapunov exponent is negative and there is a good numerical convergence of the solution in the trajectory-wise sense by using our numerical methods. Consequently, in these regimes the evolution of neuronal networks is reliable. For the chaotic dynamical regime with an intermediate strong coupling, the pseudo-Lyapunov exponent is positive, and there is no numerical convergence of the solution and only statistical quantifications of the numerical results are reliable. Finally, we present numerical evidence that the value of pseudo-Lyapunov exponent coincides with that of the standard Lyapunov exponent for systems we have been able to examine. PMID- 20020193 TI - Uncovering gamma in visual tasks. AB - Gamma is an important frequency band of the electroencephalogram (EEG), but its study has been impaired by problems with artefacts. This paper focuses on the artefacts caused by contraction of the extra-ocular muscles at the start of a saccade, which produces spurious gamma oscillations in the EEG. An algorithm was written and tested which detects and reduces the effect of this artefact. It involves novel adaptations of standard regression techniques which have traditionally been used to reduce blink artefacts, so as to render them applicable to the gamma band ocular artefact. Before the algorithm can be applied any power-line noise must be removed by noise cancellation and not notch filtering. The sharp, broadband gamma peak at around 200 ms was substantially reduced by the algorithm in all three subjects tested. However, there may be lower amplitude, task related, modulations in gamma which are uncovered when the artefact is reduced. The amplitude of the artefact had its largest positive value at the most anterior electrodes and its largest negative value at midline central and parietal electrodes, and these two sets of locations also showed the greatest reductions in gamma band magnitude after application of the algorithm. This study demonstrates the feasibility of reducing the saccade linked gamma band artefact. PMID- 20020195 TI - From the editor. PMID- 20020194 TI - HIV risks and seroprevalence among Mexican American injection drug users in California. AB - Latinos in the United States are an ethnically diverse group disproportionately affected by HIV/AIDS. We describe HIV seroprevalence, HIV risk behaviors and utilization of health services among Mexican American injection drug users (IDUs) in California (n = 286) and compare them to White (n = 830) and African American (n = 314) IDUs. Study participants were recruited from syringe exchange programs (n = 24) in California. HIV seroprevalence among Mexican Americans (0.5%) was dramatically lower than Whites (5%) and African Americans (8%). Mexican Americans reported fewer sex-related risks than Whites and African Americans though injection-related risks remained high. Compared to Whites, Mexican Americans were more likely to participate in drug treatment during a 6 month period (AOR 1.5, 95% CI 1.1, 2.0) but less likely to receive any health care (AOR 0.6, 95% CI 0.5, 0.8). Exploring cultural and structural factors among Mexican American IDUs may offer new insights into how to maintain low rates of HIV seroprevalence and reduce barriers to health care utilization. PMID- 20020196 TI - Development of a species-diagnostic marker for identification of the stingless bee Trigona pagdeni in Thailand. AB - A species-diagnostic SCAR marker for identification of the stingless bee (Trigona pagdeni Schwarz) was successfully developed. Initially, amplified fragment length polymorphism analysis was carried out across representatives of 12 stingless bee species using 64 primer combinations. A 284 bp band restrictively found in T. pagdeni was cloned and sequenced. A primer pair (CUTP1-F/R) was designed and tested for species-specificity in 15 stingless bees. The expected 163 bp fragment was successfully amplified in all examined individuals of T. pagdeni (129/129). Nevertheless, cross-species amplification was also observed in T. fimbriata (1/3), T. collina (11/112), T. laeviceps (1/12), and T. fuscobalteata (15/15), but not in other species. SSCP analysis of CUTP1 further differentiated T. fuscobalteata and T. collina from T. pagdeni. Although T. laeviceps, T. fimbriata, and T. pagdeni shared an identical SSCP genotype, they are not taxonomically problematic species. PMID- 20020197 TI - An online survival analysis tool to rapidly assess the effect of 22,277 genes on breast cancer prognosis using microarray data of 1,809 patients. AB - Validating prognostic or predictive candidate genes in appropriately powered breast cancer cohorts are of utmost interest. Our aim was to develop an online tool to draw survival plots, which can be used to assess the relevance of the expression levels of various genes on the clinical outcome both in untreated and treated breast cancer patients. A background database was established using gene expression data and survival information of 1,809 patients downloaded from GEO (Affymetrix HGU133A and HGU133+2 microarrays). The median relapse free survival is 6.43 years, 968/1,231 patients are estrogen-receptor (ER) positive, and 190/1,369 are lymph-node positive. After quality control and normalization only probes present on both Affymetrix platforms were retained (n = 22,277). In order to analyze the prognostic value of a particular gene, the cohorts are divided into two groups according to the median (or upper/lower quartile) expression of the gene. The two groups can be compared in terms of relapse free survival, overall survival, and distant metastasis free survival. A survival curve is displayed, and the hazard ratio with 95% confidence intervals and logrank P value are calculated and displayed. Additionally, three subgroups of patients can be assessed: systematically untreated patients, endocrine-treated ER positive patients, and patients with a distribution of clinical characteristics representative of those seen in general clinical practice in the US. Web address: www.kmplot.com . We used this integrative data analysis tool to confirm the prognostic power of the proliferation-related genes TOP2A and TOP2B, MKI67, CCND2, CCND3, CCNDE2, as well as CDKN1A, and TK2. We also validated the capability of microarrays to determine estrogen receptor status in 1,231 patients. The tool is highly valuable for the preliminary assessment of biomarkers, especially for research groups with limited bioinformatic resources. PMID- 20020198 TI - Expression pattern of stromal cell-derived factor-1 chemokine in invasive breast cancer is correlated with estrogen receptor status and patient prognosis. AB - Chemokine receptor CXCR4 is known to be crucially involved in tumor progression, but the role of its ligand, stromal cell-derived factor-1 (SDF-1), remains unclear. The present study was conducted to clarify the clinicopathological and prognostic impact of SDF-1 expression in invasive breast cancers. Expression of SDF-1 mRNA and protein was examined in five breast cancer cell lines with or without estradiol treatment. In 52 surgically resected breast cancers, the level of SDF-1 mRNA in frozen samples and the pattern of SDF-1 protein immunoreactivity in formalin-fixed paraffin-embedded tissue sections were compared. In another cohort of 223 breast cancers, the correlation between SDF-1 immunoreactivity and clinicopathological parameters was examined using a tissue microarray. Estradiol treatment markedly increased the expression of SDF-1 mRNA and protein in the estrogen receptor (ER)-positive cell lines, MCF-7 and T47D. Among the 52 resected breast cancers, those with a cytoplasmic-dominant pattern of SDF-1 expression showed higher SDF-1 mRNA levels (median 27.4) than those with a membrane-dominant or negative pattern (median 13.6, P = 0.0017). Accordingly, the cytoplasmic dominant pattern was defined as "high SDF-1 expression," and other patterns were defined as "low SDF-1 expression." Among the cohort of 223 tumors, "high SDF-1 expression" was detected in 158 (70.9%) and was significantly correlated with ER positivity (P < 0.0001), HER2 negativity (P = 0.021), and lower grade (P < 0.0001). Univariate analysis demonstrated that "high SDF-1 expression" was a significant indicator of better clinical outcome in both the entire patient cohort (P = 0.017) and the 133 patients with ER-positive tumors (P = 0.036), but not in the 90 patients with ER-negative tumors. Multivariate analysis showed that SDF-1 status was an independent factor related to overall survival in patients with ER-positive tumors (P = 0.046). SDF-1 status is a significant prognostic factor and may be clinically useful for assigning adjuvant therapy to patients with ER-positive invasive breast cancers. PMID- 20020199 TI - Identification of an aminopeptidase from the skeletal muscle of grass carp (Ctenopharyngodon idellus). AB - Aminopeptidases play important roles in turnover of proteins, metabolism of hormones and neurotransmission, cell maturation and immunological regulations. In the present study, an aminopeptidase was purified to homogeneity from the skeletal muscle of grass carp by ammonium sulfate fractionation and sequential chromatographic steps, including DEAE-Sephacel, Sephacryl S-200, hydroxyapatite and Phenyl-Sepharose. The purified enzyme revealed a molecular mass of approximately 105 kDa both on SDS-PAGE and on gel filtration of Superdex 200. The enzymatic activity toward synthetic substrates was optimal at 40 degrees C and pH 7.0-7.5. Metal-chelating agents such as EDTA and EGTA effectively inhibited the enzyme activity while inhibitors to serine, asparatic and cysteine proteinases did not show much effect, suggesting its belonging to metalloproteinase family. A specific aminopeptidase inhibitor bestatin was most effective in suppressing the enzymatic activity and performed in a competitive fashion. The enzymatic activity was slightly enhanced by metal ions of Mg2+ and Mn2+ while inhibited to different extents by Co2+, Cu2+, Zn2+ and Ca2+. Sulfhydryl reagent was necessary to maintain its activity. Purified enzyme demonstrated amidolytic activity most effectively against synthetic aminopeptidase substrate Leu-methylcoumarylamide (MCA) while N-terminal-blocked substrates and myofibrillar proteins were not hydrolyzed. The enzyme purified in the present study was quite possibly a leucine aminopeptidase (LAP) and functions during muscular protein metabolism. PMID- 20020200 TI - Critical role of connexin43 in zebrafish late primitive and definitive hematopoiesis. AB - In vitro studies have suggested that connexin43 (cx43) expression is of particular importance during establishment and regeneration of the mammalian hematopoietic system. However, little is known about its in vivo functions during hematopoiesis due to the embryonic lethality of mammalian knockout models. In this study, we observed that zebrafish cx43 is not only expressed in the eyes, cerebellum, heart, and vasculature, but also expressed, albeit at low levels, in intermediate cell mass (ICM, the primitive hematopoietic site). Knockdown of cx43 leads to vacuolization in the wedge of the ICM and an apparent reduction in the number of circulating blood cells, but does not affect their cellular morphology. Whole-mount in situ hybridization analysis revealed that the hemangioblastic marker flk-1 and the primitive hematopoietic markers lmo2 and scl are basically maintained at normal levels in cx43 morphant embryos at 12-13 h postfertilization (hpf) compared with the con-MO injected embryos. However, subsequent expression of the definitive hematopoietic stem cell (HSC) marker c-myb was severely downregulated in the ventral wall of the dorsal aorta of cx43-depleted embryos at 36 hpf. Furthermore, we confirmed this phenotype by injection of cx43-MO into Tg(gata1:EGFP) embryos. Together, our results show that cx43 contributes to late primitive and definitive hematopoiesis in zebrafish embryos. PMID- 20020201 TI - Distinct embryotoxic effects of lithium appeared in a new assessment model of the sea urchin: the whole embryo assay and the blastomere culture assay. AB - Early embryogenesis is one of the most sensitive and critical stages in animal development. Here we propose a new assessment model on the effect of pollutant to multicellular organism development. That is a comparison between the whole embryo assay and the blastomere culture assay. We examined the LiCl effect on the sea urchin early development in both of whole embryos and the culture of isolated blastomeres. The mesoderm and endoderm region were capable to differentiate into skeletogenic cells when they were isolated at 60-cell stage and cultured in vitro. The embryo developed to exogastrula by the vegetalizing effect of the same LiCl condition where ectodermal region changed their fate to endoderm, while the isolated blastomeres from the presumptive ectoderm region differentiated into skeletogenic cells in the culture with LiCl. The effect of LiCl to the sea urchin embryo and to the dissociated blastomere is a unique example where same cells response distinctly to the same agent depend on the condition around them. Present results show the importance of examining the process in cellular and tissue levels for the exact understanding on the morphological effect of chemicals and metals. PMID- 20020202 TI - Designing and evaluation of sodium selenite nanoparticles in vitro to improve selenium absorption in ruminants. AB - Sodium selenite is used to prevent selenium deficiency known as nutritional muscular dystrophy or white muscle disease. In ruminants, selenium supplements are transformed partiality in insoluble form by ruminal microorganisms and its process decrease the selenium absorption in digestive gastrointestinal. However, the objective in this research was focused in encapsulated sodium selenite to be release into of a pH less than four, similarity to an intestinal environment. It was encapsulated by nanoprecipitation and emulsion-evaporation methods, within polymeric nanoparticles. The effect of these methods, polymer proportion (Eudragit RL and RS) and solvent (ethanol and acetone) on the physicochemical (drug entrapment, polidispersity index (PDI) and z potential) and morphological characteristics (particle morphology and particle size) were evaluated. Particle size from each nanoparticles, formulation ranged from 36.64 to 213.86 nm. Particle size, z potential and PDI increased (P 0.05) were observed when different polymeric proportions were used. Selenium entrapment was 26% when emulsion-evaporation method was used and 78% with nanoprecipitation. Nanoparticles produced by nanoprecipitation were spherical and had a great variation in particle size; on the other hand, nanoparticles produced by emulsion evaporation were spherical as well as amorphous and presented a homogeneous nanopartcicle size distribution. The release of selenium from nanoparticles was higher in acid pH (less than 4), this condition may represent a better availability of the mineral in the small intestine. PMID- 20020203 TI - Polymorphism of BMPR1B, BMP15 and GDF9 fecundity genes in prolific Garole sheep. AB - Mutation studies in different prolific sheep breeds have shown that the transforming growth factor beta super family ligands viz. the growth differentiation factor 9 (GDF9/FecG), bone morphogenetic protein 15 (BMP15/FecX) and associated type I receptors, bone morphogenetic protein receptor (BMPR1B/FecB), are major determinant of ovulation rate and consequent increase in litter size. The Garole sheep is a highly prolific sheep breed of India. Characterization of fecundity genes in these animals could substantially improvise the breeding programme in these animals as well as other sheep breeds of the region. The present study was therefore designed with the objective of polymorphism study of fecundity genes in these prolific microsheep. A total of 11 point mutations were detected by polymerase chain reaction (PCR)-based method. A competitive technique called tetra-primer amplification refractory mutation system-PCR was adapted to type a total of ten points of two ovine fecundity genes (GDF9 and BMP15). The FecB locus of the BMPR1B gene and G1 locus of GDF9 gene were found to be polymorphic. In FecB locus, two genotypes, wild type (FecB(+)) and mutant (FecBB), were detected with allele frequencies of 0.39 and 0.61, respectively. At G1 locus, two genotypes, mutant (A) and wild types (G) were detected with allele frequencies of 0.18 and 0.82, respectively. This study reports Garole sheep as the fourth sheep breed after Belclare/Cambridge, Lacaune and Small-tailed Han sheep, where coexisting polymorphism has been found in two different fecundity genes (BMPRIB and GDF9 genes). PMID- 20020204 TI - Comparison of prominent glomerular injury and prominent tubular injury in immunoglobulin A nephropathy. AB - OBJECTIVES: The goal of this study was to define the differences between prominent glomerular injury and prominent tubular injury in immunoglobulin (Ig) A nephropathy patients presenting with acute renal failure. METHODS: In our experience of 236 patients with biopsy-proven IgA nephropathy from 2000 to 2009 at a medical center in Taiwan, 20 cases of acute renal failure were identified. The patients' records were retrospectively reviewed with respect to clinical presentation, morphology of renal biopsy, and outcomes. RESULTS: We subdivided the 20 patients into two groups: group 1 included 9 patients with prominent glomerular injury (>=30% of glomeruli affected by crescents) and group 2 included 11 patients with prominent tubular injury (tubules filled with red blood cell casts/acute tubular necrosis/or acute interstitial nephritis). Regarding clinical parameters, the glomerular injury group had a greater urinary protein excretion (P = 0.014), a higher level of peak serum creatinine (P = 0.025), and more frequently had requirement for dialysis support (P = 0.022). Concerning histological parameters, the glomerular injury group had a higher degree of mesangial hypercellularity (P = 0.009). Regarding renal outcome, the tubular injury group had a higher remission rate than the glomerular injury group (P = 0.022). CONCLUSIONS: Histological characteristics showed two distinct mechanisms of acute kidney injury associated with IgA nephropathy, including prominent glomerular injury and prominent tubular injury. The clinical presentation and prognosis largely depended on the histological presentation. PMID- 20020205 TI - Incidence of end-stage renal disease in the elderly: a steadily rising global socioeconomic epidemic. PMID- 20020206 TI - Late diagnosis of primary hyperoxaluria after failed kidney transplantation. AB - Primary hyperoxaluria type 1 (PH1) is a rare autosomal recessive inborn error of the glyoxylate metabolism that is based on absence, deficiency or mislocalization of the liver-specific peroxisomal enzyme alanine:glyoxylate aminotransferase. Hyperoxaluria leads to recurrent formation of calculi and/or nephrocalcinosis and often early end-stage renal disease (ESRD) accompanied by systemic calcium oxalate crystal deposition. In this report, we describe an adult female patient with only one stone passage before development of ESRD. With unknown diagnosis of PH, the patient received an isolated kidney graft and developed an early onset of graft failure. Although initially presumed as an acute rejection, the biopsy revealed calcium oxalate crystals, which then raised a suspicion of primary hyperoxaluria. The diagnosis was later confirmed by hyperoxaluria, elevated plasma oxalate levels and mutation of the AGXT gene, showing the patient to be compound heterozygous for the c.33_34InsC and c.508G > A mutations. Plasma oxalate levels did not decrease after high-dose pyridoxine treatment. Based on this case report, we would recommend in all patients even with a minor history of nephrolithiasis but progression to chronic renal failure to exclude primary hyperoxaluria before isolated kidney transplantation is considered. PMID- 20020207 TI - Short-term efficacy of sevelamer versus calcium acetate in patients with chronic kidney disease stage 3-4. AB - BACKGROUND: The relative effectiveness and safety of sevelamer, a mineral-free phosphate binder, for treatment of hyperphosphatemia in children with chronic kidney disease is uncertain. AIM: This study was designed to compare the efficacy and acceptability of sevelamer hydrochloride to calcium acetate as a phosphate binder in pediatric patients with chronic kidney disease. METHODS: A 12-week open label trial of sevelamer hydrochloride vs calcium acetate was initiated in 22 patients, aged 2-18, with CKD stages 3 and 4. After a 2-week washout of phosphate binders and vitamin D, patients were randomized to receive sevelamer hydrochloride or calcium acetate. The effect of therapy was adjusted for baseline blood levels of calcium, phosphorus, calcium-phosphate product, alkaline phosphatase, PTH and GFR using ANOVA. The primary end point was the decrease in serum phosphorus levels after 12 weeks of treatment. RESULTS: Of the 22 patients enrolled, data of 19 patients were used for analysis. The adjusted mean serum phosphate levels at 12 weeks did not differ significantly between calcium acetate (5.3 mg/dl) and sevelamer-treated subjects (6.1 mg/dl) (P adjusted means = 0.6). The adjusted blood level of calcium at 12 weeks was significantly lower in the sevelamer-treated patients (8.2 mg/dl) compared to those treated with calcium acetate (9.1 mg/dl) (P adjusted means = 0.01). In the sevelamer group, there was a non-significant decrease in serum bicarbonate, whereas the total and LDL cholesterol significantly decreased at 12 weeks (P = 0.04). Sevelamer hydrochloride was well tolerated and without adverse effects related to the drug. CONCLUSIONS: Compared to calcium acetate, use of sevelamer in children with chronic kidney disease is associated with similar reduction in serum phosphate levels, lower risk of hypercalcemia, and marked decrease in serum lipid levels. PMID- 20020208 TI - Benefits of family and social relationships for Thai parents living with HIV. AB - Family and social relationships are important structural supports in Thailand that are likely to influence the health and mental health of persons living with HIV (PLH). Structural equation modeling examined these relationships among 409 PLH in two communities in Thailand. Latent variables were constructed for most outcomes and mediators, with adherence to antiretroviral (ARV) therapy, depression, and disclosure represented by single-item indicators. All models controlled for gender, age, and education. Disclosure was significantly and positively associated with ARV adherence, and to both family functioning and social support. Family functioning and social support were significantly related to the PLH's self-perceptions of health and mental health, as well as being significantly correlated with each other. Better family functioning was significantly associated with better quality of life, better perceived health, fewer symptoms of depression, and greater ARV adherence. Social support was significantly associated with better quality of life and fewer depressive symptoms. These results highlight the important role that an organized and structured family life and social support network can play in encouraging better health outcomes among PLH. PMID- 20020209 TI - Effects of the Communities That Care model in Pennsylvania on change in adolescent risk and problem behaviors. AB - Despite the public health burden of adolescent substance use, delinquency, and other problem behavior, few comprehensive models of disseminating evidence-based prevention programs to communities have demonstrated positive youth outcomes at a population level, capacity to maintain program fidelity, and sustainability. We examined whether the Communities That Care (CTC; Hawkins and Catalano 1992) model had a positive impact on risk/protective factors and academic and behavioral outcomes among adolescents in a quasi-experimental effectiveness study. We conducted a longitudinal study of CTC in Pennsylvania utilizing biannual surveillance data collected through anonymous in-school student surveys. We utilized multilevel models to examine CTC impact on change in risk/protective factors, grades, delinquency, and substance use over time. Youth in CTC communities demonstrated less growth in delinquency, but not substance use, than youth in non-CTC communities. Levels of risk factors increased more slowly, and protective factors and academic performance decreased more slowly, among CTC community grade-cohorts that were exposed to evidence-based, universal prevention programs than comparison grade cohorts. Community coalitions can affect adolescent risk and protective behaviors at a population level when evidence based programs are utilized. CTC represents an effective model for disseminating such programs. PMID- 20020210 TI - LateNight Penn State alcohol-free programming: students drink less on days they participate. AB - Despite the public health importance of alcohol-free social programs for college students, the majority of existing campus strategies have not been empirically evaluated. This study utilized repeated daily reports to examine the association between attendance at campus-led alcohol-free programming and alcohol use on specific days while controlling for individuals' typical rates of use. The current study assessed students' participation in the LateNight Penn State (LNPS) alcohol-free programming and amount of alcohol use at a daily level, in order to determine whether students consumed less alcohol on days they attended LNPS compared to weekend days they did not attend. First-year college students reported their daily social activity involvement and alcohol use via 14 consecutive daily web-based surveys. Multilevel regression analyses modeled variation in alcohol use on weekend days (N = 3,350) nested within people (N = 689 people, 51% women). Analyses focused on within-individual differences between nights attending and not attending LNPS, thereby controlling for stable individual differences, measured and unmeasured. Results indicated that students drank less on days they attended LNPS and on days they stayed in (rather than going to bars/parties, other campus events, or entertainment), both especially among women. These results suggest that alcohol-free social programs may be an effective strategy for decreasing alcohol use on days when students attend alcohol-free events rather than going to other events or gatherings. PMID- 20020211 TI - Improved preservation of the tissue surrounding percutaneous devices by hyaluronic acid and dermatan sulfate in a human skin explant model. AB - Cellular apoptosis and proliferation was analyzed in an in vitro culture system of organotypic human skin explants in the presence or absence of external fixator pins. The effect on the tissues of a mixture of hyaluronic acid and dermatan sulfate (HA+DS) delivered at the skin-pin interface was also studied. After 2 weeks in culture, skin specimens interfaced with fixator pins showed increased keratinocyte apoptosis and proliferation compared to specimens without fixator pins. Simultaneously, a relative reduction of apoptosis and proliferation was observed in specimens treated with the HA+DS mixture, regardless of fixation pin presence. In addition, the HA+DS mixture appeared to help in the preservation of the epidermal basal membrane. It is concluded that in this in vitro model, fixator pins induce keratinocyte apoptosis and hyperproliferation, which are reduced in the presence of the HA+DS mixture. These methods may be useful for a better maintenance of the soft tissue surrounding percutaneous devices in vivo. PMID- 20020213 TI - Estimation of viscous dissipative stresses induced by a mechanical heart valve using PIV data. AB - Among the clinical complications of mechanical heart valves (MHVs), hemolysis was previously thought to result from Reynolds stresses in turbulent flows. A more recent hypothesis suggests viscous dissipative stresses at spatial scales similar in size to red blood cells may be related to hemolysis in MHVs, but the resolution of current instrumentation is insufficient to measure the smallest eddy sizes. We studied the St. Jude Medical (SJM) 27 mm valve in the aortic position of a pulsatile circulatory mock loop under physiologic conditions with particle image velocimetry (PIV). Assuming a dynamic equilibrium assumption between the resolved and sub-grid-scale (SGS) energy flux, the SGS energy flux was calculated from the strain rate tensor computed from the resolved velocity fields and the SGS stress was determined by the Smagorinsky model, from which the turbulence dissipation rate and then the viscous dissipative stresses were estimated. Our results showed Reynolds stresses up to 80 N/m2 throughout the cardiac cycle, and viscous dissipative stresses below 12 N/m2. The viscous dissipative stresses remain far below the threshold of red blood cell hemolysis, but could potentially damage platelets, implying the need for further study in the phenomenon of MHV hemolytic complications. PMID- 20020214 TI - Clavispora lusitaniae and Chaetomium atrobrunneum as rare agents of cutaneous infection. AB - We describe the first case of cutaneous infection caused by Chaetomium atrobrunneum and Clavispora lusitaniae in a one-and-a-half-year-old boy with acute and severe inflammation around his left eyelid. He presented to our outpatient center with a 6-day history and previously ineffective antibacterial therapy. Scanning electron microscopy (SEM) revealed hyphae and spores were on the surface of the crusty exudates and also penetrated into it, and the microbiology study further showed their characteristic cultural features. Fungal isolates were identified by the amplification and sequencing of the 26S RNA gene and of the ITS region, as C. lusitaniae and C. atrobrunneum. Up until now, most known clinical records of these rare species have shown them as agents of deep mycosis. Due to the emergency situation, medications were administered promptly and confirmed by subsequent fungal identification and successful therapeutic outcome. This article illustrates the importance of recognizing fungal infections, especially those caused by uncommon pathogens. Limitations in the routine identification procedures and therapeutic options of this emerging opportunistic agent are also discussed in this report. PMID- 20020215 TI - Comparison of the effects of systemic (intramuscular) and intrastriatal administration of a selective D1 dopamine receptor blocker on motor behavior and postural rearrangement in dogs. AB - The present study was based on the concept (Shapovalova, 2000) that activation and blockade of different types of muscarinic and dopamine receptors located in different efferent outputs of the neostriatum should have different effects on motor behavior. The aim of the study was to compare the effects of systemic and intrastriatal injections of a selective D(1) dopamine receptor blocker on motor behavior. Experiments were performed on five dogs using a model of an operant defensive reflex associated with maintaining a defined flexor posture. The experiments, with recording, storage, and analysis of data, were performed using an original personal computer program. Systemic (intramuscular) administration of the D(1) dopamine receptor blocker SCH23390 at a dose of 0.025 mg/kg led to a sharp reduction in the amplitude of the operant defensive reflex, with complete refusal to perform it in most cases. The phasic component of the operant response showed the most significant level of inhibition, though the diagonal pattern of the postural rearrangement persisted. Bilateral microinjections of the same D(1) receptor blocker into the neostriatum at doses of 0.1 and 1.0 microg did not alter the proportion of correct solutions of the operant task, though the microinjections did induce a series of changes in motor activity, with significant increases in the latent period of the response, and in some cases increases in response amplitude, decreased phasicity, and complete cessation of intersignal raisings of the limb. These data lead to the following conclusions: 1) the difference in the effects of systemic and intrastriatal administration of SCH23390 evidently results from the fact that systemic administration can also block D(1) receptors in other structures in addition to the neostriatum; 2) the effects of the nigrostriatal dopaminergic system on the neostriatum mediated by D(1) receptors are complex, with activation of motor activity (projection spiny neurons in the direct pathway) and weak modulation of mental processes (large aspiny cholinergic interneurons); 3) regulation of movement and postural rearrangement is mediated by different efferent outputs of the neostriatum. PMID- 20020216 TI - 1st UK Purine Symposium: 19th November 2009. PMID- 20020217 TI - Adenosine A(2A) agonist and A(2B) antagonist mediate an inhibition of inflammation-induced contractile disturbance of a rat gastrointestinal preparation. AB - Adenosine can show anti-inflammatory as well as pro-inflammatory activities. The contribution of the specific adenosine receptor subtypes in various cells, tissues and organs is complex. In this study, we examined the effect of the adenosine A(2A) receptor agonist CGS 21680 and the A(2B)R antagonist PSB-1115 on acute inflammation induced experimentally by 2,4,6-trinitrobenzenesulfonic acid (TNBS) on rat ileum/jejunum preparations. Pre-incubation of the ileum/jejunum segments with TNBS for 30 min resulted in a concentration-dependent inhibition of acetylcholine (ACh)-induced contractions. Pharmacological activation of the A(2A)R with CGS 21680 (0.1-10 microM) pre-incubated simultaneously with TNBS (10 mM) prevented concentration-dependently the TNBS-induced inhibition of the ACh contractions. Stimulation of A(2B)R with the selective agonist BAY 60-6583 (10 microM) did neither result in an increase nor in a further decrease of ACh induced contractions compared to the TNBS-induced inhibition. The simultaneous pre-incubation of the ileum/jejunum segments with TNBS (10 mM) and the selective A(2B)R antagonist PSB-1115 (100 microM) inhibited the contraction-decreasing effect of TNBS. The effects of the A(2A)R agonist and the A(2B)R antagonist were in the same range as the effect induced by 1 microM methotrexate. The combination of the A(2A)R agonist CGS 21680 and the A(2B)R antagonist PSB-1115 at subthreshold concentrations of both agents found a significant amelioration of the TNBS-diminished contractility. Our results demonstrate that the activation of A(2A) receptors or the blockade of the A(2B) receptors can prevent the inflammation-induced disturbance of the ACh-induced contraction in TNBS pre treated small intestinal preparations. The combination of both may be useful for the treatment of inflammatory bowel diseases. PMID- 20020218 TI - Rapid development of fasting-induced hepatic lipidosis in the American mink (Neovison vison): effects of food deprivation and re-alimentation on body fat depots, tissue fatty acid profiles, hematology and endocrinology. AB - Hepatic lipidosis is a common pathological finding in the American mink (Neovison vison) and can be caused by nutritional imbalance due to obesity or rapid body weight loss. The objectives of the present study were to investigate the timeline and characterize the development of hepatic lipidosis in mink in response to 0-7 days of food deprivation and liver recovery after 28 days of re-feeding. We report here the effects on hematological and endocrine variables, body fat mobilization, the development of hepatic lipidosis and the alterations in the liver lipid classes and tissue fatty acid (FA) sums. Food deprivation resulted in the rapid mobilization of body fat, most notably visceral, causing elevated hepatosomatic index and increased liver triacylglycerol content. The increased absolute amounts of liver total phospholipids and phosphatidylcholine suggested endoplasmic reticulum stress. The hepatic lipid infiltration and the altered liver lipid profiles were associated with a significantly reduced proportion of n 3 polyunsaturated FA (PUFA) in the livers and the decrease was more evident in the females. Likewise, re-feeding of the female mink resulted in a more pronounced recovery of the liver n-3 PUFA. The rapid decrease in the n-3/n-6 PUFA ratio in response to food deprivation could trigger an inflammatory response in the liver. This could be a key contributor to the pathophysiology of fatty liver disease in mink influencing disease progression. PMID- 20020219 TI - Erratum to: open access implementation and diabetes outcomes. PMID- 20020221 TI - Anterograde intussusception following laparoscopic Roux-en-Y gastric bypass: a case report and review of the literature. AB - Intussusception is a rare but worrisome cause of bowel obstruction in patients following Roux-en-Y gastric bypass. When intussusceptions is discovered in the general adult population, most often there is a "lead-point"; however, following bariatric procedures, this may not be true. There appears to be an increased incidence of this condition in open compared to laparoscopic Roux-en-Y gastric bypass procedures. Intussusception is often difficult to diagnose, especially in this population. Symptoms and signs can be very vague, and even computed tomography may not be accurate in diagnosing this condition. A high index of suspicion is required to successfully diagnose intussusception, and treatment often requires exploration and bowel resection. Herein, we report a case that follows several of these trends and suggests other possible contributions to intussusception. We also review other cases of intussusception after laparoscopic gastric bypass reported in the literature. PMID- 20020220 TI - Update in women's health. PMID- 20020222 TI - Asymptomatic sleep-disordered breathing in premenopausal women awaiting bariatric surgery. AB - BACKGROUND: There is a strong association between sleep-disordered breathing (SDB) and obesity. However, there are no studies addressed to determining the prevalence of SDB in morbidly obese premenopausal women, the most frequent group of patients requiring bariatric surgery. To evaluate the prevalence and characteristics of sleep apnea-hypopnea syndrome (SAHS) and obesity hypoventilation syndrome (OHS) in morbidly obese pre-menopausal women included in a program of bariatric surgery. METHODS: A total of 88 consecutive morbidly obese pre-menopausal women (38.3 +/- 8.1 years, body mass index (BMI) 48.0 +/- 6.7 kg/m(2)) being evaluated for bariatric surgery at the outpatient obesity unit of a university hospital were prospectively recruited. SDB examinations included a non-attended respiratory polygraphy, pulmonary function testing, and an awake arterial gasometry. SAHS was defined as an apnea-hypopnea index (AHI) >or=10 events per hour and patients were divided in non-SAHS (AHI <10), mild (AHI: 10 20), moderate (AHI: 21-30), and severe (AHI >30). OHS was defined as the presence of hypercapnia (PaCO(2) > 45). Somnolence was evaluated using the Epworth sleepiness scale. RESULTS: A total of 66 (75.0%) obese patients had SAHS: 25 (28.4%) mild, 14 (15.9%) moderate, and 27 (30.6%) severe. Seven (7.9%) obese patients presented OHS. Excessive daily somnolence was recorded in only 11 (16.6%) of the obese women with SAHS. In multiple regression analysis, BMI was the only variable independently associated with SAHS severity. CONCLUSIONS: Due to its high prevalence and severity, it should be recommended to investigate SDB in all morbidly obese pre-menopausal women awaiting bariatric surgery even in the absence of excessive daytime sleepiness. PMID- 20020223 TI - Production of lactic acid from raw sweet potato powders by rhizopus oryzae immobilized in sodium alginate capsules. AB - Rhizopus oryzae immobilized in calcium alginate was applied in lactic acid fermentation with unhydrolyzed raw sweet potato powders as the sole carbon source. The effects of sodium alginate concentration, calcium chloride concentration, and the immobilized bead diameter on lactic acid production were investigated. Increase in sodium alginate concentration during the gelation process would harden the immobilized capsule, which led to a decrease in lactic acid production. The increase in calcium chloride would increase the thickness of the immobilized capsule, which would increase the mass transfer resistance. Nevertheless, while the calcium chloride was lower than 15%, it would not have obvious effects on lactic acid production. A larger bead could have more space for cell growth, which led to the maximum lactic acid production observed at the 5-mm bead diameter. Moreover, results of repeated-batch operation suggested that immobilized cells could have higher stability in lactic acid production than free cells. The total cumulative lactic acid in immobilized-cell operation could increase by 55% as compared with free-cell operation after 216 h (seven repeated batches), and no loss of amylolytic activity was observed. The results indicated that immobilized R. oryzae by Ca-alginate could be suitable for lactic acid production from unhydrolyzed raw potato powders. PMID- 20020224 TI - A proof for substitution of endogenous iron (II) in lipoxygenase by exogenous Cu2+. AB - Soybean lipoxygenase (LOX) contains endogenous iron (II) at the active site, which is important for the enzyme activity. The activity of LOX can be accelerated by some exogenous metal ions including Cu2+. However, the mechanism of the activity improvement caused by exogenous metal ions remains unclear, not only for LOX but for most other metalloenzymes. Meanwhile, the possibility that exogenous metal ions can displace endogenous iron (II) is still in discussion for a lack of a direct and quantitative proof. In this paper, a quantitative proof of replacing iron (II) inside LOX by exogenous Cu2+ was provided, simply using UV Vis spectrometry with two indicators p-carboxylantipyrylazo and 9-(4 carboxyphenyl)-2,3,7-trihydroxyl-6-fluorine. A 0.56 microM free iron (II) was observed in the bulk solution after incubating 9.45 microM Cu2+ with 16.10 microM LOX at 20 degrees C for 5 min, which is in coincidence with the decrement of Cu2+ in the bulk solution (0.53 microM), implying that iron (II) was replaced by Cu2+. PMID- 20020225 TI - Simultaneous treatment of unresectable hepatocelluar carcinoma and hepatic artery aneurysm, case report. AB - INTRODUCTION: Hepatic artery aneurysm is uncommon with an estimated incidence of less than 0.25%. Because most patients are asymptomatic, the diagnosis is usually made as an incidental finding on imaging studies performed for other reasons. Because of their propensity to rupture with potential catastrophic intraperitoneal hemorrhage, early diagnosis is important. Herein, relatively asymptomatic aneurysm of the common hepatic artery mostly of atherosclerotic etiology is presented. The importance of imaging findings in the diagnosis of this condition is discussed and relevant literature is reviewed (1, 2). Hepatocellular carcinoma (HCC) ranks among the most common malignancies worldwide, and the prognosis for patients with HCC is typically poor. Chemoembolization has become the mainstay of treatment for patients with unresectable HCC. Transcatheter arterial chemoembolization is intended to deliver a highly concentrated dose of chemotherapy to tumor cells, prolong the contact time between the chemotherapeutic agents and the cancer cells, and minimize systemic toxicity. Ideally, achieving these goals will result in a tumor shrinkage, symptomatic relief, improved quality of life, and increased patient survival (3). METHODS: We will present a case of male patient, 72 years old, who was referred for transcatheter arterial chemoembolization for unresectable hepatocelluar carcinoma. RESULTS: Helical CT scan showed right lobe mass infiltrating the peritoneum with enhancement in the early arterial phase together with hepatic artery aneurysm that was successfully treated at the same time using emulsion of N-butyl cyanoacylate and lipodol with a concentration of 1:1. CONCLUSION: Hepatic artery aneurysms are uncommon lesions that have varied clinical presentations. Early diagnosis is essential because the natural tendency of the lesion is to rupture into peritoneal cavity or surrounding organs. Chemoembolization has become the mainstay of treatment for patients with unresectable HCC. Our case is notable, because atherosclerotic aneurysms of the hepatic artery are extremely rare with very few cases reported so far and to diagnose a hepatic artery aneurysm and to treat it in one setting with chemoembolization of unresectable HCC without rupture of the aneurysm is also unusual. PMID- 20020226 TI - NMR assignments of the DNA binding domain of Ml4 protein from Mesorhizobium loti. AB - Ml4 protein from Mesorhizobium loti has a 58% sequence identity with the Ros protein from Agrobacterium tumefaciens that contains a prokaryotic Cys(2)His(2) zinc finger domain. Interestingly, Ml4 is a zinc-lacking protein that does not contain the Cys(2)His(2) motif and is able to bind the Ros DNA target sequence with high affinity. Here we report the (1)H, (15)N and (13)C NMR assignments of the Ml4 protein DNA binding domain (residue 52-151), as an important step toward elucidating at a molecular level how this prokaryotic domain can overcome the metal requirement for proper folding and DNA-binding activity. PMID- 20020227 TI - Resonance assignment and secondary structure of the middle MA-3 domain and complete tandem MA-3 region of the tumour suppressor protein Pdcd4. AB - Pdcd4 (Programmed Cell Death Protein 4) is a novel eukaryotic tumour suppressor protein, which is involved in the regulation of both transcription and translation (reviewed in Lankat-Buttgereit and Goke 2009). The protein contains two interacting MA-3 domains (MA-3(M) and MA-3(C)), which are linked by a short semi-flexible linker region (Waters et al. 2007; Suzuki et al. 2008). The MA-3 domains are involved in mediating specific protein-protein interactions with functional partners such as eIF4A (Yang et al. 2003 ). Here we report essentially complete backbone and side chain (15)N, (13)C and (1)H assignments for a construct composed of the middle MA-3 domain and subsequent linker region (MA 3(M)) and backbone assignments for the entire tandem MA-3 region of Pdcd4 (Pdcd4 MA-3(M-C)). Analysis of the backbone chemical shift data obtained indicates that Pdcd4 MA-3(M) contains eight helical regions corresponding to over 74% of the protein backbone and that Pdcd4 MA-3(M-C) contains fifteen helical regions (72%). Comparison of the position of these helical regions with those observed in the crystal structures suggests that the solution and crystal structures of both proteins are very similar. PMID- 20020228 TI - Gene expression profiles of a mouse congenic strain carrying an obesity susceptibility QTL under obesigenic diets. AB - Genetic factors are strongly involved in the development of obesity, likely through the interactions of susceptibility genes with obesigenic environments, such as high-fat, high-sucrose (HFS) diets. Previously, we have established a mouse congenic strain on C57BL/6 J background, carrying an obesity quantitative trait locus (QTL), tabw2, derived from obese diabetic TALLYHO/JngJ mice. The tabw2 congenic mice exhibit increased adiposity and hyperleptinemia, which becomes exacerbated upon feeding HFS diets. In this study, we conducted genome wide gene expression profiling to evaluate differentially expressed genes between tabw2 and control mice fed HFS diets, which may lead to identification of candidate genes as well as insights into the mechanisms underlying obesity mediated by tabw2. Both tabw2 congenic mice and control mice were fed HFS diets for 10 weeks beginning at 4 weeks of age, and total RNA was isolated from liver and adipose tissue. Whole-genome microarray analysis was performed and verified by real-time quantitative RT-PCR. At False Discovery Rate adjusted P < 0.05, 1026 genes were up-regulated and 308 down-regulated in liver, whereas 393 were up regulated and 187 down-regulated in adipose tissue in tabw2 congenic mice compared to controls. Within the tabw2 QTL interval, 70 genes exhibited differential expression in either liver or adipose tissue. A comprehensive pathway analysis revealed a number of biological pathways that may be perturbed in the diet-induced obesity mediated by tabw2. PMID- 20020229 TI - Salmonella enteritidis necrotising fasciitis in a multiple myeloma patient receiving bortezomib. PMID- 20020230 TI - An unusual case of transient dermatological reaction to bortezomib in AL amyloidosis. AB - We report an unusual dermatological reaction to bortezomib in a 61-year-old man with AL amyloidosis. Systemic AL amyloidosis is a rare complication of monoclonal gammopathy or myeloma in which abnormally unstable free light chains cause fibrillary deposits in organs leading to multisystem disease. The treatment of AL amyloidosis is directed at the underlying plasma cell dyscrasia and most regimes have been adapted from myeloma, but drug toxicity is more common in AL amyloidosis because of the more extensive nature of the disease. We report a patient who developed asymptomatic purple discolouration of the veins of his left arm several days after receiving the infusion in his left hand, although the infusion itself had been uncomplicated with no extravasation. The discolouration resolved completely within 2 weeks; there was recurrence on a subsequent dose of bortezomib but this also subsided spontaneously. This reaction may have been transient phlebitis or a local vasogenic reaction; its transient nature and the lack of systemic features suggest it is a benign phenomenon. There appears to be no indication for discontinuation of bortezomib treatment or dose alteration in such cases. PMID- 20020231 TI - Prasugrel: Clinical development and therapeutic application. AB - Dual antiplatelet therapy with aspirin and clopidogrel is a cornerstone of the management of patients with acute coronary syndromes and following percutaneous coronary intervention. Despite the proven benefits, clear limitations of clopidogrel exist. Prasugrel is a third-generation thienopyridine antiplatelet agent with pharmacologic characteristics that overcome some of the limitations of clopidogrel, but at the expense of increased bleeding. The promising results seen with prasugrel in large, randomized trials led to its recent approval by the US Food and Drug Administration for reducing thrombotic cardiovascular events in patients with acute coronary syndromes managed with percutaneous coronary intervention. This article will review the limitations of standard antiplatelet therapy and discuss the clinical application of prasugrel. PMID- 20020233 TI - Changes in corneal endothelial cell density in patients with normal-tension glaucoma. AB - PURPOSE: To study the changes in corneal endothelial cell density in eyes with normal-tension glaucoma compared with those in controls. METHODS: A total of 227 subjects in three groups, one each of normal-tension glaucoma and primary open angle glaucoma patients and one of normal controls, were studied from January 2008 to July 2008 in the Ophthalmology Department of Kangbuk Samsung Hospital. The glaucoma (normal-tension glaucoma and primary open-angle glaucoma) patients included monocular and binocular glaucoma patients. Corneal endothelial cells were examined using a noncontact specular microscope. RESULTS: The mean endothelial cell densities in the three groups were as follows: normal-tension glaucoma group, 2696.7+/-303.9 cell/mm2; primary open-angle glaucoma group, 2370.5+/-392.3 cell/mm2; and normal group, 2723.6+/-300.6 cell/mm2. The endothelial cell count was not significantly different between normal-tension glaucoma and normal groups (P=1.000). Primary open-angle glaucoma patients had significantly lower endothelial cell counts (P<0.001) than the normal group. The endothelial cell count was also significantly lower in eyes with primary open angle glaucoma than in normal-tension glaucoma eyes (P<0.001). CONCLUSIONS: There was a significant decrease in corneal endothelial cell density in eyes with primary open-angle glaucoma, but not in eyes with normal-tension glaucoma. Elevated intraocular pressure likely affected the decrease of corneal endothelial cell density in eyes with glaucoma. PMID- 20020234 TI - Correlation between blue-on-yellow perimetry and scanning laser polarimetry with variable corneal compensation measurements in primary open-angle glaucoma. AB - OBJECTIVE: To investigate the correlation between blue-on-yellow perimetry (B/YP) indices and scanning laser polarimetry with variable corneal compensation (GDx VCC) parameters in primary open-angle glaucoma (POAG) patients, and to evaluate whether the two tests can detect similar glaucomatous damage. METHODS: One eye each of 86 patients with POAG was chosen. All patients were examined with the Octopus 101 automated perimetry dG2 program using the dynamic strategy/normal method (white-onwhite perimetry, W/WP), with the dG2 program using the dynamic strategy/BY method (blue-on-yellow perimetry, B/YP), and with GDx VCC. Findings were analyzed by using Pearson's correlation analysis. RESULTS: Of the 42 early glaucoma patients, 28 had both B/YP visual field loss and retinal nerve fiber layer (RNFL) defects detected by the GDx VCC measurements, and in 85.7% (24/28) of patients the regions of B/YP visual field defects corresponded to the RNFL defects detected by GDx VCC. All moderate and late glaucoma patients had both B/YP visual field loss and RNFL defects detected by GDx VCC, and in 95.5% (42/44) the regions of B/YP visual field defects corresponded to the RNFL defects detected by GDx VCC. A statistically significant correlation was found between B/YP mean sensitivity (MS) and the temporal-superior-nasal-inferior-temporal (TSNIT) average (r2>0.191, P>0.047) in the early glaucoma group, but no significant correlation was found between the W/WP indices and GDx VCC parameters. In the moderate to late glaucoma group, a low or moderate, but significant, correlation was found between B/YP MS and GDx VCC parameters (r2>0.286-0.405, P>0.001-0.01), with the most correlative parameter with B/YP MS being the TSNIT average. A low but significant correlation was also found between the B/YP mean defect (MD) and TSNIT average, superior average, inferior average, and nerve fiber indicator (NFI) (r2>0.188-0.271, P>0.013-0.044), and a low or moderate significant correlation was also found between the W/WP MS and MD and the GDx VCC parameters. CONCLUSION: The presence of significant correlations between some B/YP indices and the GDx VCC parameters in glaucoma patients suggests that the two tests can detect similar areas and amounts of glaucomatous damage. These findings raise the possibility that both techniques can be used in clinical examinations to yield convergent data. PMID- 20020235 TI - Fixation behavior in advanced stage glaucoma assessed by the MicroPerimeter MP-1. AB - PURPOSE: To investigate fixation behavior in eyes with advanced glaucoma using the MicroPerimeter MP-1. METHODS: We retrospectively reviewed 39 glaucoma patients who had scotomas adjacent to fixation points. Using the MP-1, we examined the stability and location of fixation with the fixation test and the microperimetry test. We examined retinal sensitivity using the central 10-2 SITA standard programs of a Humphrey Field Analyzer and the macula 10 degrees program of the MP-1 and analyzed the correlation between fixation behavior and retinal sensitivity. RESULTS: Of the 39 eyes, 37 showed "stable" fixation in the fixation test, while 30 eyes showed stable fixation in the microperimetry test. In the fixation test, 32 of 39 eyes demonstrated "predominantly central" fixation, whereas in the microperimetry test only 26 eyes exhibited the same fixation. Fixation stability correlated positively with sensitivity in the central 10 degrees diameter area (r=0.414, P=0.009). Among the six eyes showing "predominantly eccentric" fixation, the preferred retinal locus of five was in the superior or superotemporal direction from the fovea. CONCLUSIONS: The MP-1 illustrated the fixation patterns in glaucomatous eyes and the fixation patterns correlated well with retinal sensitivity. PMID- 20020236 TI - Photodynamic therapy for polypoidal choroidal vasculopathy: baseline perimetric results and visual outcomes. AB - PURPOSE: To investigate whether the baseline mean deviation (MD) of Humphrey perimetry has a predictive value for visual improvement after photodynamic therapy (PDT) for polypoidal choroidal vasculopathy (PCV). METHODS: We retrospectively reviewed the records of 40 eyes of 39 PCV patients (28 men, 11 women) who underwent PDT. The follow-up period ranged from 12 to 42 months. Best corrected visual acuity (BCVA) improved more than 0.2 logarithm of the minimum angle of resolution (logMAR) units at the final examination in 22 (55%) of the 40 eyes, which were classified into the "Improved group." The other 18 eyes with improvement of 0.2). One day after suprachoroidal injection, slight choroidal hemorrhage occurred. No severe intraocular reactions were observed in any of the three groups, and FFA and IA showed no edema or leakage around blood vessels. There was no significant difference in latency or amplitude of the ERG wave among the three groups (P>0.1). Histopathological examinations showed the suprachoroidal FG induced localized inflammation (from acute inflammation to chronic phagocytosis) at the early stage and finally turned into fibrosis tissue. CONCLUSIONS: FG is feasible for suprachoroidal use, and no obvious toxic reaction was observed in either in vitro or in vivo studies. PMID- 20020246 TI - Refractory eosinophilic granulation tissue of the palpebral conjunctiva. PMID- 20020247 TI - Squamous cell carcinoma of the conjunctiva invading the orbit in a non-Hodgkin lymphoma. PMID- 20020248 TI - Limbal nodules associated with Sweet's syndrome. PMID- 20020249 TI - A case of corneal endothelial deterioration associated with retained Descemet's membrane after penetrating keratoplasty. PMID- 20020250 TI - Topographical alteration in the cornea after photodynamic therapy for neovascularization in lipid keratopathy. PMID- 20020251 TI - Cladosporium cladosporioides keratomycosis: a case report. PMID- 20020252 TI - A case of atypical Cogan's syndrome with posterior scleritis and uveitis. PMID- 20020253 TI - Secondary acute angle closure with spontaneous suprachoroidal hemorrhage suspected by ultrasound biomicroscopic examination. PMID- 20020254 TI - Bilateral acute functional disturbance in the retina following placental abruption. PMID- 20020255 TI - A case of transient myopia after blunt eye trauma. PMID- 20020256 TI - Orbital inflammation caused by a spirurid larva type X showing small speckles on magnetic resonance imaging. PMID- 20020257 TI - Drug-induced liver injury. AB - Many drugs and environmental chemicals are capable of evoking some degree of liver injury. The liver represents a primary target for adverse drug reactions due to its central role in biotransformation and excretion of foreign compounds, its portal location within the circulation exposing it to a wide variety of substances, and its anatomic and physiologic structure. Drug-induced liver injury (DILI) remains the single most common adverse indication leading to drug candidate failure or withdrawal from the market. However, the absolute incidence of DILI is low, and this presents a challenge to mechanistic studies. DILI remains unpredictable making prevention very difficult. In this chapter, we focus on the current understanding of DILI. We begin with an overview regarding the significance and epidemiology of DILI and then examine the clinical presentation and susceptibility factors related to DILI. This is followed by a review of the current literature regarding the proposed pathogenesis of DILI, which involves the participation of a drug, or most often a reactive metabolite of the drug, that either directly affects cellular function or elicits an immune response. It is our hope that this chapter will shed light on the major problems associated with DILI in regards to the pharmaceutical industry, drug regulatory agencies, physicians and pharmacists, and patients. PMID- 20020258 TI - Drug hypersensitivity reactions involving skin. AB - Immune reactions to drugs can cause a variety of diseases involving the skin, liver, kidney, lungs, and other organs. Beside immediate, IgE-mediated reactions of varying degrees (urticaria to anaphylactic shock), many drug hypersensitivity reactions appear delayed, namely hours to days after starting drug treatment, showing a variety of clinical manifestations from solely skin involvement to fulminant systemic diseases which may be fatal. Immunohistochemical and functional studies of drug-specific T cells in patients with delayed reactions confirmed a predominant role for T cells in the onset and maintenance of immune mediated delayed drug hypersensitivity reactions (type IV reactions). In these reactions, drug-specific CD4+ and CD8+ T cells are stimulated by drugs through their T cell receptors (TCR). Drugs can stimulate T cells in two ways: they can act as haptens and bind covalently to larger protein structures (hapten-carrier model), inducing a specific immune response. In addition, they may accidentally bind in a labile, noncovalent way to a particular TCR of the whole TCR repertoire and possibly also major histocompatibility complex (MHC)-molecules - similar to their pharmacologic action. This seems to be sufficient to reactivate certain, probably in vivo preactivated T cells, if an additional interaction of the drug stimulated TCR with MHC molecules occurs. The mechanism was named pharmacological interaction of a drug with (immune) receptor and thus termed the p-i concept. This new concept may explain the frequent skin symptoms in drug hypersensitivity to oral or parenteral drugs. Furthermore, the various clinical manifestations of T cell-mediated drug hypersensitivity may be explained by distinct T cell functions leading to different clinical phenotypes. These data allowed a subclassification of the delayed hypersensitivity reactions (type IV) into T cell reactions which, by releasing certain cytokines and chemokines, preferentially activate and recruit monocytes (type IVa), eosinophils (type IVb), or neutrophils (type IVd). PMID- 20020259 TI - Adverse drug reactions affecting blood cells. AB - Numerous medications and other xenobiotics are capable of producing adverse reactions (ADRs) affecting red cells, platelets or neutrophils. Occasionally, more than one blood element is affected simultaneously. As with all drug reactions, some side effects are a direct consequence of a known pharmacologic action of the drug and are dose-dependent; others occur sporadically and relatively independent of dose. The latter ("idiosyncratic") reactions are unpredictable and, in general, have no known underlying genetic basis. Many are antibody-mediated, as would be expected since cellular immune effector cells have little direct access to circulating blood cells. In this chapter, we will discuss idiosyncratic drug reactions affecting blood and blood forming tissues with an emphasis on those thought to be immune-mediated. PMID- 20020260 TI - Molecular mechanisms of adverse drug reactions in cardiac tissue. AB - The myocardium is the target of toxicity for a number of drugs. Based on pharmacological evidence, cellular targets for drugs that produce adverse reactions can be categorized into a number of sites that include the cell membrane-bound receptors, the second messenger system, ionic channels, ionic pumps, and intracellular organelles. Additionally, interference with the neuronal input to the heart can also present a global site where adverse drug effects can manifest themselves. Simply, a drug can interfere with the normal cardiac action by modifying an ion channel function at the plasma membrane level leading to abnormal repolarization and/or depolarization of the heart cells thus precipitating a disruption in the rhythm and causing dysfunction in contractions and/or relaxations of myocytes. It is now recognized that toxic actions of drugs against the myocardium are not exclusive to the antitumor or the so-called cardiac drugs, and many other drugs with diverse chemical structures, such as antimicrobial, antimalarial, antihistamines, psychiatric, and gastrointestinal medications, seem to be capable of severely compromising myocardium function. At present, great emphasis in terms of drug safety is being placed on the interaction of many classes of drugs with the hERG potassium channel in cardiac tissue. The interest in the latter channel stems from the simplified view that drugs that block the hERG potassium channel cause prolongation of the QT interval, and this can cause life-threatening cardiac arrhythmias. Based on the evidence in the current literature, this concept does not seem to always hold true. PMID- 20020261 TI - Mechanisms of drug-induced nephrotoxicity. AB - Drug-induced nephrotoxicity is a common complication of several medications and diagnostic agents. It is seen in both inpatient and outpatient settings with variable presentations ranging from mild, reversible injury to advanced kidney disease. Manifestations of drug-induced nephrotoxicity include acid-base abnormalities, electrolyte imbalances, urine sediment abnormalities, proteinuria, pyuria, hematuria, and, most commonly, a decline in the glomerular filtration rate. The mechanisms of drug-induced nephrotoxicity may differ between various drugs or drug classes, and they are generally categorized based on the histological component of the kidney that is affected. Aminoglycoside antibiotics, radiocontrast media, conventional nonselective nonsteroidal anti inflammatory drugs, and selective cyclooxygenase-2 inhibitors, amphotericin B, and angiotensin-converting enzyme inhibitors have been frequently implicated. This chapter reviews the clinical presentation and basic mechanisms of drug induced nephrotoxicity. PMID- 20020262 TI - Receptor- and reactive intermediate-mediated mechanisms of teratogenesis. AB - Drugs and environmental chemicals can adversely alter the development of the fetus at critical periods during pregnancy, resulting in death, or in structural and functional birth defects in the surviving offspring. This process of teratogenesis may not be evident until a decade or more after birth. Postnatal functional abnormalities include deficits in brain function, a variety of metabolic diseases, and cancer. Due to the high degree of fetal cellular division and differentiation, and to differences from the adult in many biochemical pathways, the fetus is highly susceptible to teratogens, typically at low exposure levels that do not harm the mother. Insights into the mechanisms of teratogenesis come primarily from animal models and in vitro systems, and involve either receptor-mediated or reactive intermediate-mediated processes. Receptor mediated mechanisms involving the reversible binding of xenobiotic substrates to a specific receptor are exemplified herein by the interaction of the environmental chemical 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD or "dioxin") with the cytosolic aryl hydrocarbon receptor (AHR), which translocates to the nucleus and, in association with other proteins, binds to AH-responsive elements (AHREs) in numerous genes, initiating changes in gene transcription that can perturb development. Alternatively, many xenobiotics are bioactivated by fetal enzymes like the cytochromes P450 (CYPs) and prostaglandin H synthases (PHSs) to highly unstable electrophilic or free radical reactive intermediates. Electrophilic reactive intermediates can covalently (irreversibly) bind to and alter the function of essential cellular macromolecules (proteins, DNA), causing developmental anomalies. Free radical reactive intermediates can enhance the formation of reactive oxygen species (ROS), resulting in oxidative damage to cellular macromolecules and/or altered signal transduction. The teratogenicity of reactive intermediates is determined to a large extent by the balance among embryonic and fetal pathways of xenobiotic bioactivation, detoxification of the xenobiotic reactive intermediate, detoxification of ROS, and repair of oxidative macromolecular damage. PMID- 20020263 TI - Role of reactive metabolites in drug-induced hepatotoxicity. AB - Drugs are generally converted to biologically inactive forms and eliminated from the body, principally by hepatic metabolism. However, certain drugs undergo biotransformation to metabolites that can interfere with cellular functions through their intrinsic chemical reactivity towards glutathione, leading to thiol depletion, and functionally critical macromolecules, resulting in reversible modification, irreversible adduct formation, and irreversible loss of activity. There is now a great deal of evidence which shows that reactive metabolites are formed from drugs known to cause hepatotoxicity, such as acetaminophen, tamoxifen, isoniazid, and amodiaquine. The main theme of this article is to review the evidence for chemically reactive metabolites being initiating factors for the multiple downstream biological events culminating in toxicity. The major objectives are to understand those idiosyncratic hepatotoxicities thought to be caused by chemically reactive metabolites and to define the role of toxic metabolites. PMID- 20020264 TI - The role of cytokines in the mechanism of adverse drug reactions. AB - Cytokines are thought to play a role in acute and/or immune-mediated adverse drug reactions (ADRs) due to their ability to regulate the innate and adaptive immune systems. This role is highly complex owing to the pluripotent nature of cytokines, which enables the same cytokine to play multiple roles depending on target organ(s) involved. As a result, the discussion of cytokine involvement in ADRs is organized according to target organ(s); specifically, ADRs targeting skin and liver, as well as ADRs targeting multiple organs, such as drug-induced autoimmunity and infusion-related reactions. In addition to discussing the mechanism(s) by which cytokines contribute to the initiation, propagation, and resolution of ADRs, we also discuss the usefulness and limitations of current methodologies available to conduct such mechanistic studies. While animal models appear to hold the most promise for uncovering additional mechanisms, this field is plagued by a lack of good animal models and, as a result, the mechanism of cytokine involvement in ADRs is often studied using less informative in vitro studies. The recent formation of the Drug-Induced Liver Injury Network, whose goal is collect thousands of samples from drug-induced liver injury patients, has enormous potential to advance knowledge in this field, by enabling large-scale cytokine polymorphism studies. In conclusion, we discuss how further advances in this field could be of significant benefit to patients in terms of preventing, predicting, and treating ADRs. PMID- 20020265 TI - The keap1-nrf2 cellular defense pathway: mechanisms of regulation and role in protection against drug-induced toxicity. AB - Adverse drug reactions pose a significant public health problem. In some cases, the process of drug metabolism can contribute to the onset of toxicity through the bioactivation of a parent molecule to a chemically reactive intermediate. In order to maintain a favorable balance between bioactivation and detoxification, mammalian cells have evolved an inducible cell defense system known as the antioxidant response pathway. The activity of this cytoprotective pathway is largely regulated by the transcription factor Nrf2, which governs the expression of many phase II detoxification and antioxidant enzymes. In turn, the activity of Nrf2 is regulated by the cysteine-rich cytosolic inhibitor Keap1, which acts as a "sensor" for chemical/oxidative stress. This article summarizes our current understanding of the molecular mechanisms that regulate the function of the Keap1 Nrf2 pathway and highlights the importance of Nrf2 in the protection against drug induced toxicity. PMID- 20020266 TI - Signal transduction pathways involved in drug-induced liver injury. AB - Hepatocyte death following drug intake is the critical event in the clinical manifestation of drug-induced liver injury (DILI). Traditionally, hepatocyte death caused by drugs had been attributed to overwhelming oxidative stress and mitochondria dysfunction caused by reactive metabolites formed during drug metabolism. However, recent studies have also shown that signal transduction pathways activated/inhibited during oxidative stress play a key role in DILI. In acetaminophen (APAP)-induced liver injury, hepatocyte death requires the sustained activation of c-Jun kinase (JNK), a kinase important in mediating apoptotic and necrotic death. Inhibition of JNK using chemical inhibitors or knocking down JNK can prevent hepatocyte death even in the presence of extensive glutathione (GSH) depletion, covalent binding, and oxidative stress. Once activated, JNK translocates to mitochondria, to induce mitochondria permeability transition and trigger hepatocyte death. Mitochondria are central targets where prodeath kinases such as JNK, prosurvival death proteins such as bcl-xl, and oxidative damage converge to determine hepatocyte survival. The importance of mitochondria in DILI is also observed in the Mn-SOD heterozygous (+/-) model, where mice with less mitochondrial Mn-SOD are sensitized to liver injury caused by certain drugs. An extensive body of research is accumulating suggesting a central role of mitochondria in DILI. Drugs can also cause redox changes that inhibit important prosurvival pathways such as NF-kappaB. The inhibition of NF kappaB by subtoxic doses of APAP sensitizes hepatocyte to the cytotoxic actions of tumor necrosis factor (TNF). Many drugs will induce liver injury if simultaneously treated with LPS, which promotes inflammation and cytokine release. Drugs may be sensitizing hepatocytes to the cytotoxic effects of cytokines such as TNF, or vice versa. Overall many signaling pathways are important in regulating DILI, and represent potential therapeutic targets to reduce liver injury caused by drugs. PMID- 20020267 TI - Mitochondrial involvement in drug-induced liver injury. AB - Mitochondrial dysfunction is a major mechanism of liver injury. A parent drug or its reactive metabolite can trigger outer mitochondrial membrane permeabilization or rupture due to mitochondrial permeability transition. The latter can severely deplete ATP and cause liver cell necrosis, or it can instead lead to apoptosis by releasing cytochrome c, which activates caspases in the cytosol. Necrosis and apoptosis can trigger cytolytic hepatitis resulting in lethal fulminant hepatitis in some patients. Other drugs severely inhibit mitochondrial function and trigger extensive microvesicular steatosis, hypoglycaemia, coma, and death. Milder and more prolonged forms of drug-induced mitochondrial dysfunction can also cause macrovacuolar steatosis. Although this is a benign liver lesion in the short term, it can progress to steatohepatitis and then to cirrhosis. Patient susceptibility to drug-induced mitochondrial dysfunction and liver injury can sometimes be explained by genetic or acquired variations in drug metabolism and/or elimination that increase the concentration of the toxic species (parent drug or metabolite). Susceptibility may also be increased by the presence of another condition, which also impairs mitochondrial function, such as an inborn mitochondrial cytopathy, beta-oxidation defect, certain viral infections, pregnancy, or the obesity-associated metabolic syndrome. Liver injury due to mitochondrial dysfunction can have important consequences for pharmaceutical companies. It has led to the interruption of clinical trials, the recall of several drugs after marketing, or the introduction of severe black box warnings by drug agencies. Pharmaceutical companies should systematically investigate mitochondrial effects during lead selection or preclinical safety studies. PMID- 20020269 TI - Drug-induced liver injury in humans: the case of ximelagatran. AB - Ximelagatran was the first orally available direct thrombin inhibitor under clinical development that also reached the market. Ximelagatran was tested in an extensive clinical programme. Short-term use (<12 days) in humans including the phase III clinical trials did not indicate any hepatotoxic potential. Increased hepatic enzyme levels were first observed at a higher frequency when evaluating the long-term (>35 days) use of ximelagatran (incidence of >3x upper limit of normal (ULN) plasma ALT was 7.9%). The frequency of elevated total bilirubin levels was similar in the ximelagatran and the comparator groups. However, the combination of ALT > 3x ULN and total bilirubin > 2xULN was 0.5% among patients treated with ximelagatran and 0.1% among patients in the comparator group. Symptoms such as fever and rash potentially indicating hypersensitivity (immunologic type of reaction) were low and did not differ between ximelagatran and the comparators. The withdrawal of ximelagatran from the market and termination of the ximelagatran development program was triggered by safety data from a 35-day study, indicating that severe hepatic injury in a patient could develop after exposure to the drug has been completed and that regular liver function monitoring may not mitigate the possible risk of severe hepatic injury. As for many drugs causing liver injury, the standard preclinical toxicological studies provided no indication that ximelagatran affected hepatic functions. In addition, extensive investigations using human-based in vitro models have not been able to define mechanisms explaining the pattern of hepatic injury observed in long-term clinical trials. A pharmacogenomic study provided evidence that the ALT increases were associated with major histocompatibility complex (MHC) alleles DRB1'07 and DQA1*02 suggesting a possible immunogenic pathogenesis. This example provides important clues to the mechanism of idiosyncratic drug-induced liver toxicity. PMID- 20020270 TI - Troglitazone. AB - Troglitazone was the first thiazolidinedione antidiabetic agent approved for clinical use in 1997, but it was withdrawn from the market in 2000 due to serious idiosyncratic hepatotoxicity. Troglitazone contains the structure of a unique chroman ring of vitamin E, and this structure has the potential to undergo metabolic biotransformation to form quinone metabolites, phenoxy radical intermediate, and epoxide species. Although troglitazone has been shown to induce apoptosis in various hepatic and nonhepatic cells, the involvement of reactive metabolites in the troglitazone cytotoxicity is controversial. Numerous toxicological tests, both in vivo and in vitro, have been used to try to predict the toxicity, but no direct mechanism has been demonstrated that can explain the hepatotoxicity that occurred in some individuals. This chapter summarizes the proposed mechanisms of troglitazone hepatotoxicity based in vivo and in vitro studies. Many factors have been proposed to contribute to the mechanism underlying this idiosyncratic toxicity. PMID- 20020271 TI - Nevirapine hypersensitivity. AB - Treatment of HIV-1 infections with nevirapine is associated with skin and liver toxicity. These two organ toxicities range from mild to severe, in rare cases resulting in life-threatening liver failure or toxic epidermal necrolysis. The study of the mechanistic steps leading to nevirapine-induced skin rash has been facilitated by the discovery of an animal model in which nevirapine causes a skin rash in rats that closely mimics the rash reported in patients. The similarity in characteristics of the rash between humans and rats strongly suggests that the basic mechanism is the same in both. The rash is clearly immune-mediated in rats, and partial depletion of CD4(+) T cells, but not CD8(+) T cells, is protective. We have demonstrated that the rash is related to the 12-hydroxylation of nevirapine rather than to the parent drug. This is presumably because the 12 hydroxy metabolite can be converted to a reactive quinone methide in skin, but that remains to be demonstrated. Although the rash is clearly related to the 12 hydroxy metabolite rather than the parent drug, cells from rechallenged animals respond ex vivo to the parent drug by producing cytokines such as interferon gamma with little response to the 12-hydroxy metabolite, even when the rash was induced by treatment with the metabolite rather than the parent drug. This indicates that the response of T cells in vitro cannot be used to determine what caused an immune response. We are now studying the detailed steps by which the 12 hydroxy metabolite induces an immune response and skin rash. This animal model provides a unique tool to study the mechanistic details of an idiosyncratic drug reaction; however, it is likely that there are significant differences in the mechanisms of different idiosyncratic drug reactions, and therefore the results of these studies cannot safely be generalized to all idiosyncratic drug reactions. PMID- 20020268 TI - Mechanisms of acetaminophen-induced liver necrosis. AB - Although considered safe at therapeutic doses, at higher doses, acetaminophen produces a centrilobular hepatic necrosis that can be fatal. Acetaminophen poisoning accounts for approximately one-half of all cases of acute liver failure in the United States and Great Britain today. The mechanism occurs by a complex sequence of events. These events include: (1) CYP metabolism to a reactive metabolite which depletes glutathione and covalently binds to proteins; (2) loss of glutathione with an increased formation of reactive oxygen and nitrogen species in hepatocytes undergoing necrotic changes; (3) increased oxidative stress, associated with alterations in calcium homeostasis and initiation of signal transduction responses, causing mitochondrial permeability transition; (4) mitochondrial permeability transition occurring with additional oxidative stress, loss of mitochondrial membrane potential, and loss of the ability of the mitochondria to synthesize ATP; and (5) loss of ATP which leads to necrosis. Associated with these essential events there appear to be a number of inflammatory mediators such as certain cytokines and chemokines that can modify the toxicity. Some have been shown to alter oxidative stress, but the relationship of these modulators to other critical mechanistic events has not been well delineated. In addition, existing data support the involvement of cytokines, chemokines, and growth factors in the initiation of regenerative processes leading to the reestablishment of hepatic structure and function. PMID- 20020272 TI - Mechanisms of adverse drug reactions to biologics. AB - Biologics encompass a broad range of therapeutics that include proteins and other products derived from living systems. Although the multiplicity of target organs often seen with new chemical entities is generally not seen with biologics, they can produce significant adverse reactions. Examples include IL-12 and an anti CD28 antibody that resulted in patient deaths and/or long stays in intensive care units. Mechanisms of toxicities can be categorized as pharmacological or nonpharmacological, with most, excepting hypersensitivity reactions, associated with the interaction of the agent with its planned target. Unexpected toxicities generally arise as a result of previously unknown biology. Manufacturing quality is a significant issue relative to the toxicity of biologics. The development of recombinant technology represented the single biggest advance leading to humanized products with minimal or no contaminants in comparison to products purified from animal tissues. Nevertheless, the type of manufacturing process including choice of cell type, culture medium, and purification method can result in changes to the protein. For example, a change to the closure system for erythropoietin led to an increase in aplastic anemia as a result of changing the immunogenicity characteristics of the protein. Monoclonal antibodies represent a major class of successful biologics. Toxicities associated with these agents include those associated with the binding of the complementary determining region (CDR) with the target. First dose reactions or infusion reactions are generally thought to be mediated via the Fc region of the antibody activating cytokine release, and have been observed with several antibodies. Usually, these effects (flu-like symptoms, etc.) are transient with subsequent dosing. Although biologics can have nonpharmacologic toxicities, these are less common than with small molecule drugs. PMID- 20020273 TI - Pharmacogenetics of idiosyncratic adverse drug reactions. AB - Idiosyncratic adverse drug reactions are unpredictable and thought to have an underlying genetic etiology. With the completion of the human genome and HapMap projects, together with the rapid advances in genotyping technologies, we have unprecedented capabilities in identifying genetic predisposing factors for these relatively rare, but serious, reactions. The main roadblock to this is the lack of sufficient numbers of well-characterized samples from patients with such reactions. This is now beginning to be solved through the formation of international consortia, including developing novel ways of identifying and recruiting patients affected by these reactions, both prospectively and retrospectively. This has been led by the research on abacavir hypersensitivity - its association with HLA-B*5701 forms the gold standard of how we need to identify associations and implement them in clinical practice. Strong genetic predisposing factors have also been identified for hypersensitivity reactions such as are associated with carbamazepine, allopurinol, flucloxacillin, and statin-induced myopathy. However, for most other idiosyncratic adverse drug reactions, the genetic effect sizes have been low to moderate, although this may partly be due to the fact that only small numbers have been investigated and limited genotyping strategies have been utilized. It may also indicate that genetic predisposition will be dependent on multiple genes, with complex interactions with environmental factors. Irrespective of the strength of the genetic associations identified with individual idiosyncratic adverse drug reactions, it is important to undertake functional investigations to provide insights into the mechanism(s) of how the drug interacts with the gene variant to lead to a phenotype, which can take a multitude of clinical forms with variable severity. Such investigations will be essential in preventing the burden caused by idiosyncratic reactions, both in healthcare and in industry. PMID- 20020274 TI - The danger hypothesis applied to idiosyncratic drug reactions. AB - The danger hypothesis has had a profound effect on the way immunologists view the immune response. This hypothesis proposes that the major determinant of whether an immune response is mounted against some agent is determined by whether that agent causes some type of cell damage. Assuming that most idiosyncratic drug reactions (IDRs) are immune-mediated, this hypothesis also has the potential to explain many aspects of the mechanism of these adverse drug reactions. For example, most IDRs appear to be caused by chemical metabolites rather than the parent drug, but not all drugs that form reactive metabolites are associated with a significant incidence of IDRs. Therefore, using the danger hypothesis, one feature of a drug candidate that may predict whether it causes an IDR is whether the drug, or more likely its reactive metabolites, cause cell damage. Although the range of molecules that can act as danger signals is unknown, the most attractive candidates are high mobility group box 1 protein (HMGB1), heat shock proteins, and S100 proteins. These molecules act through the same receptors (toll like receptors) as pathogen-associated molecules that stimulate the immune system. Therefore, other environmental factors such as infections or trauma might determine which patients would be at increased risk for IDRs. Although there are examples where this appears to be the case, in most cases there are no obvious environmental factors that determine IDR risk. In addition, in animal models of immune-mediated reactions, stimulation of toll-like receptors often does not increase the immune response, and depending on the timing, it can actually be protective. Therefore, there may be additional unknown control mechanisms that are involved. A better understanding of these fundamental immune mechanisms has the potential to have a significant impact on many areas of medicine. PMID- 20020275 TI - Approaches for minimizing metabolic activation of new drug candidates in drug discovery. AB - A large body of circumstantial evidence suggests that metabolic activation of drug candidates to chemically reactive electrophilic metabolites that are capable of covalently modifying cellular macromolecules may result in acute and/or immune system-mediated idiosyncratic toxicities in humans. Thus, minimizing the potential for metabolic activation of new drug candidates during the drug discovery and lead optimization stage represents a prudent strategy to help discover and develop the next generation of safe and effective therapeutic agents. In the present chapter, we discuss the scientific methodologies that currently are available to industrial pharmaceutical scientists for assessing and minimizing metabolic activation during drug discovery, their attributes and limitations, and future scientific directions that have the potential to help advance progress in this field. We also propose a roadmap that should help utilize the armamentarium of available scientific tools in a logical way and contribute to addressing metabolic activation issues in the drug discovery setting in a rapid, scientifically appropriate, and resource-conscious manner. PMID- 20020276 TI - Explaining postnatal growth plasticity in a generalist brood parasite. AB - Selection of a particular host has clear consequences for the performance of avian brood parasites. Experimental studies showed that growth rate and fledging mass of brood parasites varied between host species independently of the original host species. Finding correlates of this phenotypic plasticity in growth is important for assessing adaptiveness and potential fitness consequences of host choice. Here, I analyzed the effects of several host characteristics on growth rate and fledging mass of the young of brown-headed cowbird (Molothrus ater), a generalist, non-evicting brood parasite. Cowbird chicks grew better in fast developing host species and reached higher fledging mass in large hosts with fast postnatal development. A potential proximate mechanism linking fast growth and high fledging mass of cowbird with fast host development is superior food supply in fast-developing foster species. So far, we know very little about the consequences of the great plasticity in cowbird growth for later performance of the adult parasite. Thus, cowbird species could become interesting model systems for investigating the role of plasticity and optimization in the evolution of growth rate in birds. PMID- 20020277 TI - The earliest occurrence of the steppe pika (Ochotona pusilla) in Europe near the Pliocene/Pleistocene boundary. AB - The steppe pika (Ochotona pusilla), a representative of the lagomorph family Ochotonidae, is restricted today to Kazakhstan and Russia. This subspecies-rich form belongs to a morphologically distinct, monospecific group of relatively small pikas, inhabiting steppe-like habitat. In the fossil record, it serves as a bioindicator of dry, grassland environment. The steppe pika was abundant and widespread in Europe during the Last Glaciation, and its unquestionable presence has been reported there since the middle Pleistocene. A new discovery of O. pusilla, from the Kielniki 3B locality (Poland), dated to the latest Pliocene, moves back the species appearance in Europe about one million years, almost coeval with its first record from the late Pliocene of Kazakhstan. Presence of such a typical steppe inhabitant as O. pusilla indicates significant climate change towards more arid, continental conditions, which started influencing Europe at 2.6 Ma. We present the first reliable record of the earliest O. pusilla entry deep into Europe in the latest Pliocene. PMID- 20020278 TI - Cancer-stromal cell interaction and tumor angiogenesis in gastric cancer. AB - Recent studies in molecular and cellular biology have shown that tumor growth and metastasis are not determined by cancer cells alone but also by a variety of stromal cells. The stroma constitutes a large part of most solid tumors, and cancer-stromal cell interaction contributes functionally to tumor growth and metastasis. Angiogenesis is the result of an imbalance between positive and negative angiogenic factors released by tumor and host cells into the microenvironment of the neoplastic tissue. In gastric cancer, tumor cells and stromal cells produce various angiogenic factors, including vascular endothelial growth factor, interleukin-8, and platelet-derived endothelial cell growth factor. The microenvironment in the gastric mucosa may also influence the angiogenic phenotype of gastric cancer. Helicobacter pylori infection increases expression of several angiogenic factors by tumor cells. Activated fibroblasts and macrophages in tumor stroma also play an important role in angiogenesis and tumor progression. We review the current understanding of cancer-stromal cell interaction as it pertains to tumor angiogenesis in gastric cancer. PMID- 20020279 TI - Effects of high dose of zoledronic acid on superficial vascular network of membranous bone sites: an intravital study on rat calvarium. AB - We evaluated the impact of high dose of zoledronic acid on the superficial vascular network parameters of a membranous bone site. During a 5-day follow-up, significant reduction of the vascular density is observed only in the treated group. INTRODUCTION: The superficial vascularization is of great importance in membranous bone-healing process. A new rat calvarium intravital model was developed to study the short-term effect of a single high dose of zoledronic acid infusion on the superficial vascularization. METHODS: Optical bone chambers were implanted in the bone tissue surface of Sprague-Dawley rats' calvarium. Nine rats were injected i.v. with 400 ug/kg of zoledronic acid (Z group), and nine rats were injected with vehicle (PSS group). A 5-day follow-up of the vascular network was made by the use of pictures analysis method. RESULTS: The vascular density significantly decreases only in Z group but there was no significant difference between groups at individual time points. The total length of the vascular network decreases significantly in Z group only (p=0.003) with a significant higher decrease at D3 (p=0.04) and D5 (p=0.02) compared with control. The vascular density related to the smaller vessels (width, 5-10 um) decreases significantly between T0 and D5 in Z group only (p=0.002). CONCLUSIONS: With the help of an original intravital animal model a significant modifications on the total length of the vascular network and the vascular density of small vessels are highlighted on a membranous bone site. PMID- 20020280 TI - Blockade of adenosine A2A receptor counteracts neuropeptide-S-induced hyperlocomotion in mice. AB - Neuropeptide S (NPS) is the endogenous ligand of a G-protein-coupled receptor named as NPSR. Behavioral effects have been recently attributed to NPS, i.e. hyperlocomotion, anxiolysis, and wakefulness. However, little is known about the mechanisms by which NPS evokes such biological actions. The present study aimed to investigate the role played by the adenosine A(2A) and A(1) receptors in hyperlocomotion induced by NPS. Spontaneous locomotion was assessed in an activity cage for 30 min in mice acutely treated with caffeine (a nonselective adenosine receptor antagonist), ZM241385 (a selective A(2A) receptor antagonist), or CPT (a selective A(1) receptor antagonist) before NPS challenge (0.1 nmol, i.c.v.), which induce hyperlocomotion in mice. The pretreatment with caffeine (3 mg/kg, i.p.), in an inactive dose per se, prevented the increase in locomotion evoked by NPS. The co-administration of NPS (0.1 nmol, i.c.v.) and ZM241385 (0.1 pmol, i.c.v.) counteracted hyperlocomotion evoked by NPS. The co-administration of NPS and CPT (0.1 pmol, i.c.v.) slightly facilitated the increase in locomotion evoked by NPS alone. In summary, the pharmacological blockade of A(2A) receptors significantly attenuated the stimulatory effects of NPS. By contrast, the antagonism of A(1) receptors facilitated NPS-induced hyperlocomotion in mice, but we cannot rule out a merely additive effect of two stimulatory systems in the brain. Altogether, this is the first evidence of a putative role played by A(2A) and A(1) receptors in modulating hyperlocomotion induced by NPS. PMID- 20020281 TI - Acute rhabdomyolysis during treatment with amisulpride and metformin. PMID- 20020282 TI - A fenugreek seed extract selectively reduces spontaneous fat intake in overweight subjects. AB - PURPOSE: Fenugreek seeds (Trigonella foenum-graecum L.) have long been used as a herbal medicine for treating metabolic and nutritive dysfunctions. They have been shown to modulate feeding behaviour in animals. We have recently observed a selective decrease in fat consumption in healthy normal weight volunteers treated with a hydro-alcoholic seed extract. However, strong clinical data on the effects of fenugreek seeds on energy intake are lacking, especially in overweight individuals. The aim of our study was to investigate the effects of a repeated administration of a fenugreek seed extract on the eating behaviour of overweight subjects. METHODS: Thirty-nine healthy overweight male volunteers completed a 6 week double-blind randomized placebo-controlled parallel trial of a fixed dose of a fenugreek seed extract. Main endpoints were energy intake (dietary records and meal test), weight, fasting and post-absorptive glucose and insulin, appetite/satiety scores and oxidative parameters. RESULTS: Daily fat consumption, expressed as the ratio fat reported energy intake/total energy expenditure (fat REI/TEE), was significantly decreased in our overweight subjects administered the fenugreek seed extract relative to those receiving the placebo (fat-REI/TEE 0.26 +/- 0.02 vs. 0.30 +/- 0.01, respectively; P = 0.032). We also observed a significant decrease in the insulin/glucose ratio in subjects treated with fenugreek seed extract relative to the placebo group (0.89 +/- 0.09 vs. 1.06 +/- 0.10 mUI mmol(-1), respectively; P = 0.044). No significant effect was observed on weight, appetite/satiety scores or oxidative parameters. CONCLUSION: The repeated administration of a fenugreek seed extract slightly but significantly decreased dietary fat consumption in healthy overweight subjects in this short term study. PMID- 20020284 TI - Rhabdomyoma causing severe left ventricular outflow obstruction in a newborn: a management dilemma. PMID- 20020283 TI - Genetic determinants of acenocoumarol and phenprocoumon maintenance dose requirements. AB - OBJECTIVE: The variability in warfarin dose requirement is attributable to genetic and environmental factors. Acenocoumarol (AC) and phenprocoumon (PC) are coumarin derivates widely prescribed in European countries for the prevention and treatment of thromboembolic events. The aim of our study was to investigate the contribution of genes involved in the vitamin K cycle to AC and PC maintenance doses. METHODS: Common single nucleotide polymorphisms (SNPs) in the genes encoding cytochrome P450 family member 2C9 (CYP2C9), vitamin K epoxide reductase complex subunit 1 (VKORC1), gamma-glutamyl carboxylase (GGCX), calumenin (CALU) and apolipoprotein E (APOE) were studied in 206 patients receiving AC or PC. RESULTS: Compared to patients with the VKORC1 C1173C genotype, maintenance doses for AC or PC were reduced to 74.6 or 70.2% in heterozygous C1173T subjects and to 48.6 or 48.1% in homozygous T1173T subjects (P < 0.0001). Furthermore maintenance doses for AC and PC were significantly lower in heterozygous CYP2C9*1*3, CYP2C9*2*3, and in CYP2C9*3*3 homozygote individuals compared to homozygous CYP2C9*1*1 subjects (P = 0.0004 and P = 0.0017, respectively). A multiple regression model including age, sex, last INR, VKORC1, and CYP2C9 genotypes explained ~50% of the variability in AC/PC dose requirements. CALU genotype combinations showed minor effects on PC dose requirements. No associations with AC or PC dose requirements were observed for sequence substitutions in the GGCX or APOE genes. CONCLUSION: These results reveal that interindividual variability in weekly AC and PC maintenance dose requirement is mainly dependent on the VKORC1 1173C>T and the CYP2C9*3 alleles. VKORC1 and CYP2C9 genotyping might provide helpful information to prevent serious bleeding events in subjects receiving AC or PC. PMID- 20020285 TI - Isolation and genetic manipulation of the antibiotic down-regulatory gene, wblA ortholog for doxorubicin-producing Streptomyces strain improvement. AB - Cross-genome comparative transcriptome analyses were previously conducted using the sequenced Streptomyces coelicolor genome microarrays to understand the genetic nature of doxorubicin (DXR) and daunorubicin (DNR) overproducing industrial mutant (OIM) of Streptomyces peucetius. In this previous work, a whiB like putative transcription factor (wblA ( sco )) was identified as a global antibiotic down-regulator in S. coelicolor (Kang et al., J Bacteriol 189:4315 4319, 2007). In this study, a total genomic DNA library of a DXR/DNR overproducing S. peucetius OIM was constructed and screened using wblA ( sco ) as a probe, resulting in the isolation of a wblA ortholog (wblA ( spe )) that had 95% amino acid identity to wblA ( sco ). Gene disruption of wblA ( spe ) from the S. peucetius OIM resulted in an approximately 70% increase in DXR/DNR productivity, implying that the DXR/DNR production in the S. peucetius OIM could be further improved via comparative transcriptomics-guided target gene manipulation. Furthermore, several putative wblA ( spe ) -dependent genes were also identified using S. coelicolor interspecies DNA microarray analysis between the S. peucetius OIM and wblA ( spe )-disrupted S. peucetius OIM. Among the genes whose expressions were significantly stimulated in the absence of wblA ( spe ), the overexpression of a conserved hypothetical protein (SCO4967) further stimulated the total production of DXR/DNR/akavinone by 1.3-fold in the wblA ( spe )-disrupted S. peucetius OIM, implying that the sequential genetic manipulation of target genes identified from interspecies comparative microarray analysis could provide an efficient and rational strategy for Streptomyces strain improvement. PMID- 20020286 TI - Real-time contrast-enhanced ultrasonography of resected and immunohistochemically proven hepatic angiomyolipomas. AB - PURPOSE: The purpose of this study was to assess real-time contrast-enhanced ultrasound in patients with hepatic angiomyolipomas with respect to contrast enhancing kinetics and enhancement patterns. MATERIALS AND METHODS: Nonlinear, low mechanical index (mechanical index less than 0.2), real-time contrast enhanced ultrasonography was performed in 18 patients with 19 tumors after 2.4 mL bolus injection of contrast agent SonoVue. All the 19 tumors were surgically resected and immunohistochemically proven sporadic hepatic angiomyolipomas. RESULTS: Inhomogeneous hyperenhancing pattern was detected 16 tumors (84.2%) and homogeneous hyperenhancing pattern in 1 tumor (0.5%) in arterial phase and portal phase, 16 lesions remained hyperenhancement but one lesion became isoenhancement in late phase. Two lesions (10.5%) were detected inhomogeneously hypoenhancement compared to liver parenchyma during arterial, portal, and late phases, with only punctiform internal enhancement. The margin of all the lesions were smooth and well defined on contrast-enhanced ultrasonography, whereas no peripheral nodular arterial enhancement, centripetal filling, or spoke-wheel like enhancement pattern were depicted. Early strong enhancement in arterial phase, rapid washout of contrast agent, and appear hypoechoic to surrounding liver tissue during portal or late phase was not at all observed. CONCLUSION: Real-time contrast enhanced ultrasonography can demonstrate typical imaging characteristics of hepatic angiomyolipomas in most cases, that is, inhomogeneous hyperenhancing pattern in arterial phase, prolonged hyperenhancement during portal and late phase with smooth and well-defined margin. This real-time dynamic imaging technique may therefore improve noninvasive characterization and differentiation of this rare, benign, hepatic mesenchymal neoplasm. PMID- 20020287 TI - Aged wine and symphonic perfection. PMID- 20020288 TI - Association of increasing surgeon age with decreasing in-hospital mortality after coronary artery bypass graft surgery. AB - BACKGROUND: The aim of this study was to investigate the relation between surgeon age and in-hospital mortality for patients who underwent a coronary artery bypass graft (CABG) using a nationwide population-based data set. METHODS: This study used data from the 2004 Taiwan National Health Insurance Research Database. The study sample comprised 3766 patients hospitalized for CABG surgery and was divided into three equal-sized surgeon age groups: <40, 40 to 45, and >45 years. A conditional (fixed-effect) logistic regression was performed to examine the relation between surgeon age and in-hospital mortality after adjusting for surgeon CABG caseload and characteristics of patients and surgeons as well as the clustering effect among surgeons. RESULTS: Patients who underwent CABG performed by surgeons in the <40 years age group had significantly higher in-hospital mortality rates (5.4%) than those operated on by surgeons in the 40- to 45-year age group (3.5%) and surgeons in the >45-year age group (2.6%). Regression shows that the adjusted odds ratio of in-hospital mortality for patients operated on by surgeons in the <40-year age group was 1.47 (p < 0.05) times that for surgeons in the 40- to 45-year age group and 1.82 (p < 0.05) times that for surgeons in the >45-year age group. CONCLUSION: We conclude that older surgeons are more likely to achieve better clinical performance with CABG surgery because of their greater clinical experience. PMID- 20020290 TI - An observational trial for papillary thyroid microcarcinoma in Japanese patients. AB - BACKGROUND: The recent development and spread of ultrasonography and ultrasonography-guided fine needle aspiration biopsy (FNAB) has facilitated the detection of small papillary microcarcinomas of the thyroid measuring 1 cm or less (PMC). The marked difference in prevalence between clinical thyroid carcinoma and PMC detected on mass screening prompted us to observe PMC unless the lesion shows unfavorable features, such as location adjacent to the trachea or on the dorsal surface of the thyroid possibly invading the recurrent laryngeal nerve, clinically apparent nodal metastasis, or high-grade malignancy on FNAB findings. In the present study we report comparison of the outcomes of 340 patients with PMC who underwent observation and the prognosis of 1,055 patients who underwent immediate surgery without observation. METHODS: Between 1993 and 2004, 340 patients underwent observation and 1,055 underwent surgical treatment without observation. These 1,395 patients were enrolled in the present study. Observation periods ranged from 18 to 187 months (average 74 months). RESULTS: The proportions of patients whose PMC showed enlargement by 3 mm or more were 6.4 and 15.9% on 5-year and 10-year follow-up, respectively. Novel nodal metastasis was detected in 1.4% at 5 years and 3.4% at 10 years. There were no factors related to patient background or clinical features linked to either tumor enlargement or the novel appearance of nodal metastasis. After observation 109 of the 340 patients underwent surgical treatment for various reasons, and none of those patients showed carcinoma recurrence. In patients who underwent immediate surgical treatment, clinically apparent lateral node metastasis (N1b) and male gender were recognized as independent prognostic factors of disease-free survival. CONCLUSIONS: Papillary microcarcinomas that are not associated with unfavorable features can be candidates for observation regardless of patient background and clinical features. If there are subsequent signs of progression, such as tumor enlargement and novel nodal metastasis, it would not be too late to perform surgical treatment. Even though the primary tumor is small, careful surgical treatment including therapeutic modified neck dissection is necessary for N1b PMC patients. PMID- 20020289 TI - Mechanical compression versus subcutaneous heparin therapy in postoperative and posttrauma patients: a systematic review and meta-analysis. AB - BACKGROUND: The risk of postoperative venous thromboembolic disease is as high as 30%, with an associated fatality risk of 1%. Therefore, prophylaxis is essential, but the optimal regimen remains controversial. This study was designed to systematically review and quantitatively summarize the impact of mechanical compression versus subcutaneous heparin on venous thromboembolic disease and posttreatment bleeding in postsurgical and posttrauma patients. METHODS: Computerized searches of the MEDLINE and EMBASE databases through November 2008 were performed and supplemented with manual searches. We included studies that had: (1) a patient population undergoing surgery or admitted immediately posttrauma, (2) a randomized comparison of prophylaxis with mechanical compression versus subcutaneous heparin, (3) outcome measured in terms of deep vein thrombosis (DVT), pulmonary embolism (PE), or bleeding. RESULTS: Two reviewers independently extracted data from the original articles, which represented 16 studies, including a total of 3,887 subjects. Meta-analysis was performed using a random effects model. The pooled relative risk for mechanical compression compared with subcutaneous heparin was 1.07 (95% confidence interval [CI] 0.72, 1.61) for DVT and 1.03 (95% CI 0.48, 2.22) for PE. Mechanical compression was associated with a significantly reduced risk of postoperative bleeding compared with subcutaneous heparin (risk ratio 0.47; 95% CI 0.31, 0.70). Subgroup analyses by heparin type suggested that low molecular weight heparin may reduce risk of DVT compared with compression (relative risk 1.80; 95% CI 1.16, 2.79) but remains similarly associated with an increased risk of bleeding. CONCLUSIONS: These results suggest that the overall bleeding risk profile favors the use of compression over heparin, with the benefits in term of venous thromboembolic disease prophylaxis being similar between groups. Subgroup analyses suggest that low molecular weight heparin may have a differential effect; this observation should be further evaluated in future studies. PMID- 20020291 TI - Papillary carcinoma located in the thyroid isthmus. AB - BACKGROUND: The purpose of the present study was to evaluate the clinicopathologic characteristics and short-term outcomes of papillary thyroid carcinomas (PTC) located in the isthmus compared to tumors located in other thyroid regions, and to use those findings to establish a surgical strategy for treating these tumors. PATIENTS AND METHODS: Thyroidectomy was performed in 1,973 thyroid cancer patients in our hospital between January 2006 and December 2007. The patients were analyzed in two groups; group I, patients with tumors located in the isthmus (n = 181) and group II, patients with tumors located in other thyroid regions (n = 1,792). The clinicopathologic characteristics and short-term outcomes were compared between the two groups. RESULTS: The study groups were similar in terms of age and gender ratio. For groups I and II, respectively, the mean primary tumor size was 1.02 cm and 1.13 cm (p = 0.017), capsular invasion frequency was 70.2 and 60.8% (p = 0.008), and multifocality frequency was 48.6 and 39.8% (p = 0.006). In addition, central node involvement was 40.3 and 42.1% (p = 0.350), and lateral node involvement was 9.4 and 18.2% (p = 0.001). At the time of postoperative iodine-131 remnant ablation, 34.3% of group I patients and 38.0% of group II patients showed undetectable serum thyroglobulin concentrations. There were no significant differences in surgical complications between the two groups. CONCLUSIONS: Papillary thyroid carcinomas located in the isthmus was more likely to be associated with multiple foci in bilateral lobes and higher rates of capsular invasions than tumors in other thyroid regions. These findings indicate that total thyroidectomy is an appropriate initial surgical procedure for the isthmic PTC. PMID- 20020292 TI - Long-term outcome of differentiated thyroid carcinoma: experience in a developing country. AB - BACKGROUND: The presentation and outcome of differentiated thyroid carcinoma (DTC) in developing countries are different from the developed nations. We report the clinicopathologic profile and long-term outcome of DTC in an iodine-deficient area (IDA) in a developing country. METHODS: This retrospective study included 302 patients with DTC operated between 1989 and 2002. These patients had been followed up for a minimum period of 5 years after surgery. Clinicopathological profile, intervention, and follow-up details were noted. RESULTS: Mean age of the patients was 42 +/- 14 years. Mean follow-up period was 80 +/- 34 (24-196) months. Papillary thyroid carcinoma (PTC), follicular thyroid carcinoma (FTC), and poorly differentiated thyroid carcinoma (PDTC) was present in 62, 30, and 8% patients, respectively. Mean tumor size was 3.5 cm. Tumor multicentricity was noted in 40% of PTC, 22.2% of FTC, and 25% of PDTC patients. Lymphadenopathy was observed in 45, 10, and 67% patients with PTC, FTC, and PDTC, respectively. Extrathyroidal invasion and distant metastasis were observed in 36.8% (PTC 33%; FTC 36%; PDTC 71%) and 27% (PTC 17%; FTC 44%; PDTC 42%) of cases, respectively. Twenty percent of patients had synchronous metastases. Risk stratification ratio was 1:1.8 (high-risk vs. low-risk). Initial operative procedure was total thyroidectomy in 86.5% cases, and therapeutic lymph node dissection was performed in 37% cases. A total of 77.2% patients received adjuvant radioiodine therapy. Disease recurred in 26.6% of patients (thyroid bed recurrence 1.7%), and 21.2% patients died during follow-up. Overall survival (OS) rate at 10 years in both low-risk and high-risk groups of FTC (80 and 54%) was inferior to PTC (94 and 62%). Five-year OS for PDTC was 50%. Tumor multicentricity was a significant risk factor for OS in the low-risk group, whereas the presence of skeletal metastases and extrathyroidal invasion were significant factors for OS in the high-risk group. CONCLUSIONS: Advance stage at presentation and proportionately high rates of FTC and PDTC contribute to poor outcome of DTC in developing countries. Despite dismal outcome, total thyroidectomy seems to prevent thyroid bed recurrence in surviving patients. PMID- 20020293 TI - Role of growth factors on human parathyroid adenoma cell proliferation. AB - INTRODUCTION: Primary hyperparathyroidism (pHPT) is caused by a single monoclonal adenoma in more than 80% of patients. Biomolecular mechanisms causing pHPT are still not completely known, even if a great amount of studies have been developed recently, mainly regarding angiogenesis and growth factors. Among the latter, insulin-like growth factor 1 (IGF-1), basic fibroblastic growth factor (bFGF), vascular endothelial growth factor (VEGF), and transforming growth factor beta 1 (TGF-beta1) and their effects have been extensively evaluated in different kinds of endocrine disease. METHODS: Parathyroid cell cultures were prepared from six human adenomatous parathyroid glands that were surgically removed. After 7 days of culture, the cells were refed with DMEM supplemented with 2% FCS alone (control group), or containing hrTGFbeta1, or hrIGF-I, or hrbFGF, or hrVEGF. Then, after 48-hour incubation, cell count was performed by a particle count and size analyzer, and prevalence of cell cycle was analyzed by using a flow cytometer. RESULTS: Cell count (x10000) in the control group was 3.73 +/- 0.32. Low-dose TGF-beta1 stimulation resulted in 5.25 +/- 0.38 cells, and high-dose TGF beta1 stimulation resulted in 2.35 +/- 0.37 cells. IGF-1 stimulation resulted in 5.4 +/- 0.65 cells, bFGF stimulation in 5.68 +/- 0.86 cells, and VEGF stimulation resulted in 6.03 +/- 1.03 cells. Statistical analysis revealed significant differences in the control group compared with the growth factor-stimulated groups. Cytometry showed different results in the percentage of cells in S-phase, in particular 22.65 +/- 4.98% of IGF-1-stimulated cells were found in S-phase compared with 7.55 +/- 3.2% of control group cells (p < 0.0001). CONCLUSIONS: Growth factors seem to play an important role in parathyroid adenoma cell proliferation; IGF-1, bFGF, VEGF, and low-dose TGF-beta1 promote cell proliferation, whereas high-dose TGF-beta1 inhibits these phenomena. PMID- 20020294 TI - Morbidity and mortality of 109 consecutive cytoreductive procedures with hyperthermic intraperitoneal chemotherapy (HIPEC) performed at a community hospital. AB - BACKGROUND: Cytoreductive surgery with hyperthermic intraperitoneal chemotherapy (HIPEC) is playing an increasing role in the management of isolated peritoneal dissemination of gastrointestinal malignancies. Historically this surgery is associated with a high morbidity and mortality. Recognizing this, our study was developed to prospectively evaluate morbidity and mortality after cytoreductive surgery with HIPEC performed at a community hospital. METHODS: From January 19, 2005 to January 9, 2008, 109 consecutive patients successfully underwent cytoreductive surgery with HIPEC for peritoneal surface malignancies. All cases were performed by a single surgeon at a 323-bed community hospital. Using an institutional review board approved study we prospectively evaluated postoperative complications using the standard National Institutes of Health morbidity and mortality grading system. RESULTS: There was no 30-day or inpatient mortality. Overall grade III and IV morbidity was 30.2% (33 of 109 patients); 29.3% of the patients had at least one grade III complication, with the most common being postoperative anemia requiring a blood transfusion in 20 of the 109 patients (18.3%). Eight patients (7.3%) had wound infections, and three patients (2.7%) developed pneumonia. One patient required computerized tomography guided drainage of a pelvic abscess. There were 3 (2.7%) grade IV complications, with only one patient requiring reoperation. CONCLUSIONS: Recent studies at tertiary medical centers have shown acceptable morbidity and mortality with this procedure. Our study demonstrates that this procedure can be safely performed in the community setting as well if surgeons, other medical professionals, and ancillary caregivers have great experience in this procedure. PMID- 20020295 TI - Ratio between metastatic and examined lymph nodes (N ratio) may have low clinical utility in gastric cancer patients treated by limited lymphadenectomy: results from a single-center experience of 526 patients. AB - BACKGROUND: The aim of this study was to evaluate the prognostic value of the ratio of metastatic to examined lymph nodes (N ratio) in gastric cancer patients who underwent limited lymphadenectomy and had a small number (< or =15) of analyzed nodes. METHODS: The prognostic value of the actual AJCC/UICC pN staging system and the N ratio (0%, 1-25%, > 25%) were analyzed by means of univariate and multivariate analyses for 526 patients who underwent R0 resection for gastric adenocarcinoma at the Latvia Oncology Center. RESULTS: The mean (SD) number of analyzed nodes was 5.6 (2.8). The number of positive nodes significantly increased with the number of analyzed nodes (p < 0.001). No significant differences in survival (p = 0.508) and risk of death (p = 0.224) were observed between pN1 and pN2 subsets. When the N ratio (1-25% vs. > 25%) was taken into account, a significant difference was demonstrated between pNR1 and pNR2 with respect to survival (p = 0.017) and risk of death (p = 0.012). Nonetheless, the joint allocation of the two classifications demonstrated that only a minority of patients (28 cases) belonged to the pNR1 subset and none of these belonged to the AJCC/UICC pN2 subset. CONCLUSIONS: When a small number of lymph nodes are analyzed, the N ratio can discriminate patients better than TNM classification. However, because a small number of retrieved nodes produced only a small number of pNR1 patients, the N ratio classification cannot be justified for clinical use. PMID- 20020296 TI - Fusion technique for liver transection with Kelly-clysis and harmonic technology. AB - BACKGROUND: Various devices are available for liver transection and comparative data on transection techniques are limited by the diversity of operative procedures. Clamp crushing (Kelly-clysis) with a Cavitron ultrasonic surgical aspirator (CUSA-Integra Radionics) is widely used for splitting the liver parenchyma. Hemostasis is achieved by bipolar coagulation, ligatures, or hemoclips. We introduce a fusion technique (Focus-clysis) for liver transection using a combination of Kelly-clysis and harmonic technology. MATERIALS AND METHOD: A fusion technique (FT) was performed using FOCUS, a Kelly clamp like instrument attached to a Harmonic generator. Hepatic resections (nine major, nine minor) were performed with the fusion technique in 18 non-cirrhotic patients. Variables evaluated were blood loss, transection time, biliary leak, postoperative liver function, morbidity, and cost-effectiveness. The results were compared with 18 hepatic resections (nine major, nine minor) that were performed with our earlier technique, i.e., CUSA with bipolar cautery, ligatures, and hemoclips. RESULTS: The mean blood loss was 416 ml in the FT group, compared to 833 ml in the CUSA group. Two patients in the FT group needed blood transfusion in the first 48 h, whereas eight patients in the CUSA group had transfusions. No major postoperative liver dysfunction was noted with the new technique, and postoperative morbidity was lower in the FT group. Liver transection with the fusion technique was faster. CONCLUSION: The fusion technique (Focus-clysis) using Kelly-clysis and harmonic technology is promising, easy, and effective for liver transection. Our initial results show advantages over the routinely used CUSA/bipolar combination. The fusion technique could be a new option for liver transection, especially in non-cirrhotic patients. PMID- 20020297 TI - Risk factors for clinical pancreatic fistula after distal pancreatectomy: analysis of consecutive 100 patients. AB - BACKGROUND: The mortality associated with distal pancreatectomy (DP) has declined to <5% in recent years in high-volume centers. However, morbidity remains high, ranging from 32% to 57%. Pancreatic fistula (PF) is the most common complication after DP. The aim of this study was to analyze factors associated with the occurrence of clinical PF. METHODS: A retrospective review was performed of the medical records of 100 patients who underwent DP in our institution between May 2001 and January 2009. RESULTS: There was no mortality, but morbidity was occurred in 65 patients (65%), with major complications occurring in 9 patients (9%). PF occurred in 50 patients (50%) and clinical PF occurred in 23 patients (23%). Multivariate analysis indicated that independent risk factors for clinical PF were: age younger than 65 years (P = 0.049; odds ratio (OR) 2.958; 95% confidence interval (CI) 1.007-8.688), not ligating the main pancreatic duct (MPD) (P = 0.02; OR 4.933; 95% CI 1.283-18.967), and extended lymphadenectomy (P = 0.008; OR 4.773; 95% CI 1.504-15.145). CONCLUSIONS: Age < 65 years, not ligating the MPD, and extended lymphadenectomy are independent risk factors for clinical PF. PMID- 20020298 TI - Quantitative comparison of the difficulty of performing laparoscopic colectomy at different tumor locations. AB - BACKGROUND: Laparoscopic approaches of colectomy for colonic cancer are increasingly surpassing the mainstream open colectomy approach. Impact of disease variables, such as tumor location, has not been adequately measured in quality improvement initiatives. Quantitative analysis concerning the difficulty performing these procedures and differences in postoperative care depending on tumor site will contribute to the development of training programs and to the assessment of quality of care strategies. METHODS: A total of 3,765 cases received laparoscopic colectomy (LC). Patient demographics, weighted comorbidities, procedure-related complications, stapling devices, operating room (OR) time, postoperative length of hospital stay (LOS), or total charges (TC) were categorized and compared based on tumor location: cecum to ascending, transverse, descending, and sigmoid colon. Multivariate analyses determined the impact of tumor location on postoperative LOS, TC, OR time, and complications. RESULTS: Sigmoid colon was the most frequent tumor placement (40.5%). Significant differences in age, gender, frequency of blood transfusion, use of stapling devices, OR time, and postoperative LOS were observed among tumor locations. Transverse colon was the most significant determinant of postoperative LOS and TC, whereas descending colon tumors correlated with increased OR time. Greater OR time was associated with more postoperative resource use and complications. CONCLUSIONS: Tumor location, complications, and OR time affected postoperative resource use, whereas greater OR time signified an increased occurrence of complications. Developers of LC training programs or healthcare policy makers should consider the quantitative impact of tumor locations when attempting to improve effective skill training or to survey the quality of LC performance. PMID- 20020299 TI - Deferred primary anastomosis versus diversion in patients with severe secondary peritonitis managed with staged laparotomies. AB - BACKGROUND: There is inconclusive data on whether critically ill individuals with severe secondary peritonitis requiring multiple staged laparotomies may became eligible candidates for deferred primary anastomoses (DPA). We sought to compare a protocol for DPA against a protocol for diversion in severely ill critical patients with intra-abdominal sepsis. METHODS: A retrospective cohort study was performed examining 112 patients admitted through an ICU between 2002 and 2006, with diagnosis of secondary peritonitis and managed with staged laparotomies whom required small- or large-bowel segment resections. Patients were categorized and compared according to the surgical treatment necessitated to resolve the secondary peritonitis (DPA versus diversion). Outcome measures were days on mechanical ventilation, days required in ICU, days required in hospital, incidence of fistulas/leakages, acute respiratory distress syndrome (ARDS), and mortality. RESULTS: There were 34 patients subjected to DPA and 78 to diversion. Fistulas/leakages developed in three patients (8.8%) with DPA and four patients (5.1%) with diversion (p = 0.359). ARDS was present in 6 patients (17.6%) with DPA and 24 patients (30.8%) with diversion (p = 0.149). There were 30 patients (88.2%) with DPA and 65 patients (83.3%) with diversion discharged alive (p = 0.51). There were not statistical significant differences between groups among survivors regarding hospital length of stay, ICU length of stay, and days on mechanical ventilation. CONCLUSIONS: We did not find significant differences in morbidity or mortality when we compared DPA versus diversion surgical treatment. It is feasible to perform a primary anastomosis in critically ill patients with severe secondary peritonitis managed with staged laparotomies. PMID- 20020300 TI - Occupational radiation protection in interventional radiology: a joint guideline of the Cardiovascular and Interventional Radiology Society of Europe and the Society of Interventional Radiology. PMID- 20020301 TI - Inhibition of Streptococcus pyogenes biofilm formation by coral-associated actinomycetes. AB - Streptococcus pyogenes biofilms tend to exhibit significant tolerance to antimicrobials during infections. We screened coral-associated actinomycetes (CAA) for antibiofilm activity against different biofilm forming M serotype of Streptococcus pyogenes. Actinomycetes isolated from the mucus of the coral Acropora digitifera were screened for antibiofilm activity against S. pyogenes biofilms wherein several isolates clearly demonstrated antibiofilm activity. The biofilm inhibitory concentrations (BICs) and the sub-BICs (1/2 and 1/4 BIC) of the extracts significantly prevented biofilm formation up to 60-80%. The extract of Streptomyces akiyoshinensis (A3) displayed efficient antibiofilm activity against all the biofilm forming M serotypes. All the five extracts efficiently reduced the cell surface hydrophobicity (a crucial factor for biofilm formation in S. pyogenes) of three M types and thus may inhibit biofilm formation. CAA represent an interesting source of marine invertebrates-derived antibiofilm agents in the development of new strategies to combat Streptococcal biofilms. PMID- 20020302 TI - Differential effects of distinct bacterial biofilms in a cave environment. AB - Current microbial surveys using molecular methods provide us with critical information on the major components of natural bacterial communities. However, limited investigation has been performed on the influence of bacterial metabolism on the environment. In this study, we analyzed the pH generated by distinct bacterial communities in a cave environment. Different bacterial biofilms developing on the walls of the cave were visually distinguished by their colorations (e.g., white, yellow, and gray) and mineral depositions, and previous studies have reported on their bacterial diversity and distribution. Using pH microelectrodes, we carried out in situ measurements and were able to detect differences among these bacterial biofilms. White biofilms and carbonate depositions resulted in alkaline pH values. Gray biofilms also increased the pH although these values remained lower than in white biofilms. A combination of gray-white biofilms resulted in alkaline pH values with highest values at the white edge of the colonies. Yellow biofilms generated a slightly acid pH. These results suggest that different bacterial communities can lead to distinct effects on their environment, for instance, precipitation or dissolution of carbonates in caves. These results add information about metabolic response to current knowledge from bacterial diversity surveys, providing information on the interaction between complex bacterial communities and the geological substrate. PMID- 20020303 TI - Comparative protein modeling, prediction of conserved residue and active sites in cold resistant protein isolated from CRPF(1), a cold tolerant mutant of Pseudomonas fluorescens. AB - Proteins interacting with the biological information molecules DNA and RNA play important cellular roles in all organisms. One widespread super family of proteins implicated in such function(s) is cold shock protein (CSP) that contains the cold shock domain (CSD). This work is planned to study the three-dimensional structure, conserved residues, and different active sites in the structure of cold resistant protein (CRP) from CRPF(1), cold tolerant mutant of Pseudomonas fluorescence by comparative homology modeling. Here we tried to identify crucial residues that are involved in active sites or functional sites of the protein. The study reveals that CRP represent the prototype of the CSD and share a highly similar overall fold consisting of five antiparallel beta-sheets forming a beta barrel structure with surface exposed aromatic and basic residues that were responsible for nucleic acid binding properties of variable binding affinities and sequence selectivity and harbors the nucleic acid binding motifs RNP1 and RNP2 that is highly conserved in CSP family. PMID- 20020304 TI - [Simulation of corneal epithelial injuries by mechanical and corrosive damage : Influence of fetal bovine serum and dexpanthenol on epithelial regeneration in a cell culture model]. AB - INTRODUCTION: The present study describes simulation of corneal epithelial injury and its regeneration using an in-vitro model of immortalized human corneal epithelial cells (HCE-T) growing as monolayer cultures. MATERIAL AND METHODS: The epithelial model was damaged using defined strengths by mechanical injury or partial damage using chemical detergents (SDS and acidified medium) and subsequently the epithelium was further cultivated using serum-containing and serum-free medium supplemented with varying concentrations of calcium pantothenat. After mechanical injury wound healing was evaluated using a photomicroscope over a period of up to 48 h whereas after chemical injury a cell viability assay was used to detect the course of ATP levels in the cell layers as an indicator for the metabolic activity. RESULTS: Depending on the kind of injury pantothenat showed a regeneration enhancing effect in the concentration range from 0.001% to 0.01%. However, a concentration of 0.1% pantothenat appeared to be regeneration inhibiting. The combination of pantothenat and serum was more beneficial for wound healing than pantothenat alone, whereas serum partly levelled the effect of pantothenat. CONCLUSION: The described model allowed simulation of corneal epithelial injury and its regeneration, whereby the influence of the serum content and the kind of injury could be determined. PMID- 20020305 TI - Cardioselective nitric oxide synthase 3 gene transfer protects against myocardial reperfusion injury. AB - Nitric oxide modulates the severity of myocardial ischemia-reperfusion (I/R) injury. We investigated whether cardioselective nitric oxide synthase 3 (NOS3) gene transfer could confer myocardial protection against I/R injury in pigs and examined potential molecular mechanisms. I/R injury was induced by balloon occlusion of the left anterior descending artery for 45 min followed by 4 or 72 h reperfusion. Hemodynamic and pathological changes were measured in pigs in the absence (n = 11) or presence of prior intracoronary retroinfusion of human NOS3 (AdNOS3, 5 x 10(10) PFU, n = 13) or control vector (AdRR5, 5 x 10(10) PFU, n = 11). Retrograde NOS3 gene transfer selectively increased NOS3 expression and NO bioavailability in the area at risk (AAR) without changing endogenous NOS isoform expression. At 4 h R, LV systolic (dP/dt(max)) and diastolic (dP/dt(min)) function was better preserved in AdNOS3- than in AdRR5-injected pigs (2,539 +/- 165 vs. 1,829 +/- 156 mmHg/s, and -2,781 +/- 340 vs. -2,062 +/- 292 mmHg/s, respectively, P < 0.05 for both). Myocardial infarct size (% AAR) was significantly smaller in AdNOS3 than in control and AdRR5 and associated with a significantly greater reduction in cardiac myeloperoxidase activity, a marker of neutrophil infiltration. The latter effects were sustained at 72 h R in a subset of pigs (n = 7). In the AAR, intercellular endothelial adhesion molecule-1 expression and cardiomyocyte apoptosis were significantly lower in AdNOS3. In conclusion, single myocardial NOS3 retroinfusion attenuates I/R injury, and causes a sustained reduction in myocardial infarct size and inflammatory cell infiltration. Gene-based strategies to increase NO bioavailability may have therapeutic potential in myocardial I/R. PMID- 20020307 TI - On the origins of cold-induced vasodilation. PMID- 20020306 TI - Reduced gray matter volume of Brodmann's Area 45 is associated with severe psychotic symptoms in patients with schizophrenia. AB - Previous literature has suggested an important role of inferior frontal gyrus, which mainly consists of Brodmann's Area (BA) 44 and 45, in the pathophysiology of schizophrenia. While recent neuroimaging techniques have revealed differential functional correlates of BA 44 and 45 in healthy individuals, previous studies have not yet separately evaluated the gray matter volume reduction of BA 44 and 45 and their relationships to psychotic symptoms in patients with schizophrenia. In the present study, magnetic resonance images were obtained from 29 right handed male patients with schizophrenia and from 29 age- and handedness-matched healthy male controls. The reliable manual tracing methodology was employed to measure the gray matter volume of BA 44 and BA 45. The severities of psychotic symptoms were evaluated using the five-factor model of positive and negative syndrome scale in the patient group. A significant gray matter volume reduction of both the BA 44 and BA 45 was found bilaterally in the patients with schizophrenia compared with the healthy controls. Among these inferior frontal sub-regions, reduced volume of right BA 45 revealed the largest effect size. In addition, the reduced volume of BA 45 in left hemisphere showed a significant association with the increased severity of delusional behavior, while the severity of disorganized and positive symptoms were correlated with the bilateral BA 45 volumes in the patient group. The findings support an important role of inferior frontal gyrus in the pathophysiology of schizophrenia. The present study further demonstrated that BA 45 might especially contribute to the production of psychotic symptoms in the patients with schizophrenia. PMID- 20020308 TI - Actin-depolymerizing factor of second-generation merozoite in Eimeria tenella: clone, prokaryotic expression, and diclazuril-induced mRNA expression. AB - Actin depolymerizing factor (ADF) is an essential actin-binding protein that plays a key role in the control of actin dynamics and actin-based motility processes in intracellular parasites. To determine the effects of diclazuril on ADF gene of second-generation merozoites (mz-ADF) mRNA expression in Eimeria tenella, mz-ADF gene was cloned by RT-PCR from extracted RNA in second-generation merozoite of E. tenella and successfully expressed by pET-28a vector in Escherichia coli BL21(DE3). Results showed that the full length of the cloned cDNA sequence of the mz-ADF gene is 476 bp including an ORF of 375 bp. The sequence has 100% homology with a published sequence of sporozoite stage E. tenella ADF mRNA (GenBank EF195234.1). The recombinant protein was induced to be expressed by 1 mM isopropyl beta-D: -1-thiogalactopyranoside in vitro. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis showed that 16.99 kDa fusion protein existed in solvable form. Compared with the infected/control group, mz-ADF mRNA expression level was downregulated by 63.86% in the infected/treatment group with the treatment of diclazuril. In conclusion, the data presented here indicate that mz-ADF gene participates in an important role in the invasion host of E. tenella. Downregulation of mz-ADF mRNA expression enrich the mechanism study of diclazuril on E. tenella. PMID- 20020309 TI - [Coming to light. The problem of identification and grouping of pain patients]. PMID- 20020310 TI - Structure-based and shape-complemented pharmacophore modeling for the discovery of novel checkpoint kinase 1 inhibitors. AB - Checkpoint kinase 1 (Chk1), a member of the serine/threonine kinase family, is an attractive therapeutic target for anticancer combination therapy. A structure based modeling approach complemented with shape components was pursued to develop a reliable pharmacophore model for ATP-competitive Chk1 inhibitors. Common chemical features of the pharmacophore model were derived by clustering multiple structure-based pharmacophore features from different Chk1-ligand complexes in comparable binding modes. The final model consisted of one hydrogen bond acceptor (HBA), one hydrogen bond donor (HBD), two hydrophobic (HY) features, several excluded volumes and shape constraints. In the validation study, this feature shape query yielded an enrichment factor of 9.196 and performed fairly well at distinguishing active from inactive compounds, suggesting that the pharmacophore model can serve as a reliable tool for virtual screening to facilitate the discovery of novel Chk1 inhibitors. Besides, these pharmacophore features were assumed to be essential for Chk1 inhibitors, which might be useful for the identification of potential Chk1 inhibitors. PMID- 20020311 TI - The theoretical and experimental study on dicalcium phosphate dehydrate loading with protocatechuic aldehyde. AB - The aim of this study is to investigate the interaction between dicalcium phosphate dihydrate (CaHPO(4) x 2H(2)O, DCPD) and Protocatechuic aldehyde (C(7)H(6)O(3), Pca), which is the water-soluble constituents of Chinese Medicine, Salvia Miltiorrhiza Bunge (SMB), by calculating the absorption energy through molecular dynamics simulation. Furthermore, the effects of functional groups of Pca and temperature on Pca adsorbed by DCPD are calculated respectively. DCPD/Pca and DCPD were analyzed by X-ray diffraction (XRD), Fourier transform infrared spectroscopy (FTIR) and thermogravimetric analysis (TG). The simulation results showed that Pca mostly absorbed on the (0 2 0) surface of DCPD. The aldehyde group of Pca played a moren important role on the adsorption of Pca on DCPD than hydroxyl did, while temperature had no distinct effects on the adsorption. XRD results indicated that Pca induced the preferential growth of (0 2 0) crystal surface in DCPC/Pca whereas it had no influence on the crystal structure, the crystallinity and grain size of DCPD. FTIR and TG results showed that the characteristic peak of Pca was at 1295 cm(-1) and the content of Pca in DCPD was 16%, respectively. The present results show that molecular dynamics simulation is a very effective and complementary method to study the interaction between materials and medicine. PMID- 20020312 TI - Design and methods of a strategic outcome study for chronic kidney disease: Frontier of Renal Outcome Modifications in Japan. AB - BACKGROUND: The continuous increase in the number of people requiring dialysis is a major clinical and socioeconomical issue in Japan and other countries. This study was designed to encourage chronic kidney disease (CKD) patients to consult a physician, enhance cooperation between nephrologists and general practices, and prevent the progression of kidney disease. METHODS: Subjects comprise CKD patients aged between 40 and 74 years consulting a general physician, and patients in CKD stage 3 with proteinuria and diabetes or hypertension. This trial is a stratified open cluster-randomized study with two intervention groups: group A (weak intervention) and group B (strong intervention). We have recruited 49 local medical associations (clusters) in 15 different prefectures, which were classified into four regions (strata) based on the level of increase rate of dialysis patients. The patients in group A clusters were instructed initially to undergo treatment in accordance with the current CKD treatment guide, whereas patients in group B clusters were not only instructed in the same fashion but also received support from an information technology (IT)-based system designed to help achieve the goals of CKD treatment, consultation support centers, and consultations by dietitians visiting the local general practice offices. We assessed the rates of continued consultation, collaboration between general practitioners and nephrologists, and progression of CKD (as expressed by CKD stage). CONCLUSION: Through this study, filling the evidence-practice gap by facilitating effective communication and supporting general physicians and nephrologists, we will establish a CKD care system and decrease the number of advanced-stage CKD patients. PMID- 20020313 TI - Cryotherapy decreases histamine levels in the blood of patients with rheumatoid arthritis. AB - INTRODUCTION: Conventional physiotherapy (electrotherapy, magnetic fields), kinesitherapy, and whole-body cryotherapy (plus kinesitherapy) are used to relieve pain and inflammation or to improve function in rheumatic diseases. The aim of this study was to investigate the effects of different physiotherapies and cryotherapy on biochemical blood parameters of patients with rheumatoid arthritis (RA) and osteoarthritis (OA). MATERIALS AND METHODS: Twenty patients with RA and 17 patients with OA received whole-body cryotherapy at -140 to -160 degrees C for 2 to 3 min, once daily for 4 weeks. The second group of patients (24 with RA and 28 with OA) received conventional physiotherapy for 4 weeks. We measured the parameters of neutrophil activation (respiratory burst, calprotectin) and markers of cartilage metabolism [N-acetyl-beta-D-hexosaminidase (NAHase), ectonucleotide pyrophosphohydrolase (NTPPHase)] twice: before and 3 months after cryotherapy or physiotherapy. RESULTS: We showed, for the first time, that cryotherapy significantly reduced (P < 0.001) histamine levels in the blood of patients with RA. The effect was long-lasting (for at least 3 months). The levels of blood histamine in patients with OA were not changed significantly. Cryotherapy also downregulated the respiratory burst of PMNs and NAHase activity and upregulated calprotectin levels and the activity of NTPPHase. However, these changes were not statistically significant. In contrast, there were no significant changes in histamine levels or the other biochemical parameters measured in groups of patients treated only with physiotherapy and kinesitherapy. CONCLUSION: It may be concluded that the beneficial clinical effects of cryotherapy in RA patients are in part due to the action on the production, release, or degradation of histamine. PMID- 20020314 TI - Acute gastric lesions induced by the administration of histamine to rats with partial vascular occlusion: evidence for the gastroprotective effect of prostaglandin. AB - INTRODUCTION: Histamine is not only a potent stimulator of gastric acid secretion, but it also plays a central role in gastroduodenal ulcerogenesis. In the present study we tested the effect of pre-treatment with exogenous prostaglandin E(2) (PGE(2)) in a new rat model of experimental gastric ulcers induced by combination of histamine and gastric ischemia. METHODS: In male Wistar rats, a chronic ischemia of gastric mucosa was induced via the clamping of the left gastric artery and vein (L-AV) in combination with pylorus ligation. The following treatment groups of rats (6 rats/group) were investigated: 1) histamine alone (40 mg/kg twice s.c.); 2) vehicle (saline) followed 30 min later by gastric mucosal L-AV ischemia and pylorus ligation combined with histamine (40 mg/kg twice s.c.) and 3) PGE(2) (5 microg/kg i.g.) followed 30 min later by gastric mucosal L-AV ischemia combined with histamine (40 mg/kg twice s.c.) and pylorus ligation. At 4 hr after the clamping of L-AV and pylorus ligation, the area of gastric lesions and gastric acid secretion was determined. RESULTS: Histamine treatment failed to produce gastric lesions, but when it was combined with ischemia, the widespread gastric lesions in the corpus mucosa, but not in the antrum, were observed. This damaging effect and decrease in the GBF were significantly attenuated by pretreatment with PGE(2). CONCLUSION: The present study demonstrates that gastric hypersecretion induced by histamine in combination with gastric mucosal ischemia results in gastric lesions which progress into chronic gastric ulcers. PMID- 20020315 TI - UR-60427, a novel H4 receptor-inverse agonist that shows good efficacy in a rat asthma model. PMID- 20020316 TI - Immunological identification of the mouse H4 histamine receptor on spinal cord motor neurons using a novel anti-mouse H4R antibody. AB - In this report, we describe the development of the first anti-mouse H(4) receptor antibody. Utilising this new immunological probe, new evidence is provided for oligomeric mH(4)Rs and the presence of H(4) receptors on a subpopulation of murine motor neurons. This expands the cell types where the H(4) receptor is expressed in the mammalian CNS. PMID- 20020317 TI - Novel metabolic pathway for salicylate biodegradation via phenol in yeast Trichosporon moniliiforme. AB - A novel metabolic pathway was found in the yeast Trichosporon moniliiforme WU 0401 for salicylate degradation via phenol as the key intermediate. When 20 mM salicylate was used as the sole carbon source for the growth of strain WU-0401, phenol was detected as a distinct metabolite in the culture broth. Analysis of the products derived from salicylate or phenol through reactions with resting cells and a cell-free extract of strain WU-0401 indicated that salicylate is initially decarboxylated to phenol and then oxidized to catechol, followed by aromatic ring cleavage to form cis-cis muconate. PMID- 20020318 TI - A new fluorescent chemosensor detecting Co2+ and K+ in DMF buffered solution. AB - A new fluorescent chemosensor 2-(2-thiophene)imidazo [4,5,f]-1,10-phenanthroline (L) was prepared and characterized. By adding univalent or divalent metal ions such as Na(+), K(+), Mg(2+), Ba(2+), Mn(2+), Fe(2+), Co(2+), Ni(2+), Cu(2+), Ag(+), Zn(2+), Cd(2+) and Hg(2+) ions into the solution of L in DMF under buffered conditions with the working pH ranging from 7.0 to 8.0, we found that L could be used to detect K(+) ratiometricly and it could also be applied to sense Co(2+) with the phenomenon of fluorescence quenching of L. While the response behavior of L was not discernibly affected by other examined metal ions. PMID- 20020319 TI - Randomized controlled trial for early intervention for autism: a pilot study of the Autism 1-2-3 Project. AB - We piloted a 2-week "Autism-1-2-3" early intervention for children with autism and their parents immediately after diagnosis that targeted at (1) eye contact, (2) gesture and (3) vocalization/words. Seventeen children were randomized into the Intervention (n = 9) and Control (n = 8) groups. Outcome measures included the Autism Diagnostic Observation Schedule, Ritvo-Freeman Real Life Rating Scale, Symbolic Play Test, and Parenting Stress Index. Children with autism improved in language/communication, reciprocal social interaction, and symbolic play. Parents perceived significant improvement in their children's language, social interaction, and their own stress level. This intervention can serve as short term training on communication and social interaction for children with autism, and reduce the stress of their parents during the long waiting time for public health services. PMID- 20020320 TI - The impact of delayed blood centrifuging, choice of collection tube, and type of assay on 25-hydroxyvitamin D concentrations. AB - Studies have examined the associations between cancers and circulating 25 hydroxyvitamin D [25(OH)D], but little is known about the impact of different laboratory practices on 25(OH)D concentrations. We examined the potential impact of delayed blood centrifuging, choice of collection tube, and type of assay on 25(OH)D concentrations. Blood samples from 20 healthy volunteers underwent alternative laboratory procedures: four centrifuging times (2, 24, 72, and 96 h after blood draw); three types of collection tubes (red top serum tube, two different plasma anticoagulant tubes containing heparin or EDTA); and two types of assays (DiaSorin radioimmunoassay [RIA] and chemiluminescence immunoassay [CLIA/LIAISON((R))]). Log-transformed 25(OH)D concentrations were analyzed using the generalized estimating equations (GEE) linear regression models. We found no difference in 25(OH)D concentrations by centrifuging times or type of assay. There was some indication of a difference in 25(OH)D concentrations by tube type in CLIA/LIAISON((R))-assayed samples, with concentrations in heparinized plasma (geometric mean, 16.1 ng ml(-1)) higher than those in serum (geometric mean, 15.3 ng ml(-1)) (p = 0.01), but the difference was significant only after substantial centrifuging delays (96 h). Our study suggests no necessity for requiring immediate processing of blood samples after collection or for the choice of a tube type or assay. PMID- 20020321 TI - Selenium in surface and irrigation water in the Kendrick irrigation district, Wyoming. AB - High selenium (Se) concentrations have been found in surface waters in the Kendrick Reclamation Project, Wyoming. Precipitation and irrigation water moving over seleniferous soils are contributing causes, and drought may exacerbate this. This study surveyed Se concentrations and discharges in local surface streams, irrigation drains, and the delivery canal. Sites were sampled monthly and analyzed for Se and total suspended solids (TSS). A completely randomized design with two factors (soil parent material and location, inside or outside irrigation district) was used. Mean Se concentrations were 64 MUg L(-1) inside the irrigation district on shale soils, 17 MUg L(-1) inside the district off shale soils, 5 MUg L(-1) outside the district on shale soils, and 3 MUg L(-1) outside the district off shale soils. Correlations between discharge and Se concentrations were generally negative, while correlations between discharge and Se load were generally positive. There was little correlation between load and concentration, and little correlation between TSS and Se. A comparison of Se concentrations in streams and drains showed Se concentrations were significantly higher (p<0.001) in streams during the irrigation season, but not in the off season (p=0.515). We conclude that higher discharges decrease Se concentration, but increase load. Conversion from flood to sprinkle irrigation may increase Se concentrations by reducing discharge, but decrease Se loads going into the N. Platte River, and will likely alter the timing and magnitude of flows. Both load and concentration should be considered when implementing Se regulations and standards. PMID- 20020322 TI - Evaluation of pesticide contamination in Dilek National Park, Turkey. AB - National parks are used worldwide as a means to protect the ecological integrity of unique ecosystems. Dilek National Park in western Turkey is a protected habitat for several endangered and severely threatened species. Thirty-seven water and 59 sediment samples were collected and analyzed for pesticides at two different sampling depths. The park is contaminated with 16 different organochlorine pesticides, with more pesticides detected in sediments than in water. The most prevalent pesticides in the 30-60-cm depth were DDT (69.5% of the samples), heptachlor (62.3%), a-endosulfan (55% of samples), and endrin (37%). Lindane isomers were present, but at <30% of the sediment samples. These pesticides could be a long-term contamination source that enters the food web used by the very species the park is trying to protect. PMID- 20020323 TI - Estimation of heavy metals in commonly used medicinal plants: a market basket survey. AB - Popularity of herbal drugs is increasing all over the world because of lesser side effects as compared to synthetic drugs besides it cost effectiveness and easy availability to poor people particularly in developing countries. Keeping in view the increased market demand of herbal drugs, it is essential to ensure their chemical quality prior to use. Raw drugs and herbs are usually collected from different places, which might be contaminated with various contaminants. It is pertinent to estimate the levels of heavy metals and other micronutrients, which could be affected by their presence in the surrounding environments. Heavy metals are known to pose a potential threat to terrestrial and aquatic biota. Keeping this in view, samples of ten plants or plant parts used in drug making were collected from local markets of Punjab for heavy metal and micronutrient estimation. It was found that the samples were contaminated having cadmium, lead, chromium, iron, manganese, and zinc. The highest mean level of cadmium (23.1 MUg/kg) was found in Haritaki sample. Chromium concentration of the plant samples ranged between 7.25 and 1.34 MUg/kg with the highest values was in Daruharidra and lowest in Pippali. The levels of these heavy metals were within permissible limits. PMID- 20020324 TI - Environmental performance evaluation and strategy management using balanced scorecard. AB - Recently, environmental protection and regulations such as WEEE, ELV, and RoHS are rapidly emerging as an important issue for business to consider. The trend of swinging from end-of-pipe control to product design, green innovation, and even the establishment of image or brand has affected corporations in almost every corner in the world, and enlarged to the all modern global production network. Corporations must take proactive environmental strategies to response the challenges. This study adopts balanced scorecard structure and aim at automobile industries to understand the relationships of internal and external, financial and non-financial, and outcome and driving factors. Further relying on these relationships to draw the "map of environment strategy" to probe and understand the feasibility of environmental performance evaluation and environmental strategy control. PMID- 20020325 TI - Hypothalamic proline rich polypeptide regulates hematopoiesis. AB - The AGAPEPAEPAQPGVY proline-rich polypeptide (PRP-1) was isolated from neurosecretory granules of the bovine neurohypophysis; it is produced by N. supraopticus and N. paraventricularis. It has been shown that PRP-1 has many potentially beneficial biological effects including immunoregulatory, hematopoietic, antimicrobial and anti-neurodegenerative properties. Here we demonstrated that PRP-1 administration influence on redistribution of monocytes, granulocytes and lymphocytes between bone marrow (BM) and peripheral blood and promotes the influx of granulocytes and monocytes/macrophages from BM into peripheral blood and accumulation of immature granulocyte and monocyte in BM and delayed the maturation of T cells in BM. PRP-1 increased colony-forming cell proliferation in rat cells in vivo. In PRP-treated rat BM, the CFU number at day 4, 7 and 14 was considerably increased in comparison with untreated rats BM and no difference was found at day 21 and day 28. We found that PRP-1 enhances erythroid and myeloid colonies formation in human CD34(+) progenitor cell culture in the presence of different growth factors and down-regulates T cells colony formation and specific surface markers expression during induction of human CD34(+) progenitor cells differentiation into T lymphocytes lineage. We suggested that the hypothalamic PRP-1 possibly represents an endogenous peptide whose primary functions are to regulate neuronal survival and differentiation and hematopoiesis within neurosecretory hypothalamus-bone marrow humoral axis. PMID- 20020326 TI - Survival of native Pseudomonas in soil and wheat rhizosphere and antagonist activity against plant pathogenic fungi. AB - Survival of Pseudomonas sp. SF4c and Pseudomonas sp. SF10b (two plant-growth promoting bacteria isolated from wheat rhizosphere) was investigated in microcosms. Spontaneous rifampicin-resistant mutants derived from these strains (showing both growth rate and viability comparable to the wild-strains) were used to monitor the strains in bulk soil and wheat rhizosphere. Studies were carried out for 60 days in pots containing non-sterile fertilized or non-fertilized soil. The number of viable cells of both mutant strains declined during the first days but then became established in the wheat rhizosphere at an appropriate cell density in both kinds of soil. Survival of the strains was better in the rhizosphere than in the bulk soil. Finally, the antagonism of Pseudomonas spp. against phytopatogenic fungi was evaluated in vitro. Both strains inhibited the mycelial growth (or the resistance structures) of some of the phytopathogenic fungi tested, though variation in this antagonism was observed in different media. This inhibition could be due to the production of extracellular enzymes, hydrogen cyanide or siderophores, signifying that these microorganisms might be applied in agriculture to minimize the utilization of chemical pesticides and fertilizers. PMID- 20020327 TI - Comparison of image quality of 64-slice multidetector CT coronary CT angiography using automated and manual multiphase methods for the determination of optimal phases for image reconstruction in patients with various mean heart rates. AB - This study aimed to compare the effects of the automated and manual methods for determining the optimal phase of image reconstruction with the use of 64-slice coronary CT angiography (CCTA). In 63 patients, the optimal trigger delay was determined using an automated technique (PhaseXact) and by manual phase selection from the images reconstructed at a 10 ms and 1% increment of the R-R interval. The image quality was evaluated using four-step grading. The mean heart rate in all the patients was 70 +/- 10.1 bpm (range: 52-98 bpm). The frequency of obtaining diagnostic quality images was highest with the manual absolute timing method (97.6% of 753 segments), while it was 94.7% with manual relative timing and 91.9% with an automated method. The mean image quality score (1.3 +/- 0.2) based on the manual absolute timing was significantly better (P = 0.002, one-way ANOVA; P < 0.05, Scheffe's test) than the scores determined with the use of the other two techniques (manual relative timing, 1.4 +/- 0.3; automated method, 1.5 +/- 0.4). The best quality coronary CT images were obtained using image reconstruction with manual absolute timing for the ECG-trigger delay, while an automated method and manual relative timing provided diagnostic quality images in most of the segments. PMID- 20020328 TI - IL-9: basic biology, signaling pathways in CD4+ T cells and implications for autoimmunity. AB - CD4(+) T cell subsets play an important role in the adaptive immune response in human autoimmune diseases and in animal models of autoimmunity. In recent years, our knowledge of CD4(+) T cell differentiation has increased significantly, and new subsets continue to be recognized. Of significant importance is the recent discovery of Th9 cells, the CD4 + T cell subset that produces Interleukin-9. IL-9 has largely been regarded as a Th2 cytokine; however, it is now known that under specific conditions, Tregs, Th1, Th17 and the Th9 subset of T cells also produce IL-9. The STAT family of proteins plays a major role in the signaling pathways of these CD4(+)T subsets. Biological actions of IL-9 and the STATs signaling pathways in autoimmune diseases are continuing to be clarified. Investigation of IL-9-producing CD4(+)T cells, and elucidation of the mechanisms of IL-9-induced STATs signaling, in concert with other transcription factors, will provide a better understanding of the pathogenesis of various autoimmune diseases. PMID- 20020330 TI - Vitamin E induces phosphatidylserine externalization and red cell adhesion to endothelial cells. AB - Red blood cell (RBC) adhesion to vessel wall endothelium is a potent catalyst of vascular occlusion and occurs in oxidative stress states such as hemoglobinopathies and cardiovascular conditions. These are often treated with vitamin E (VitE), a "classic" antioxidant. In this study, we examined the effects of VitE on RBC adhesion to vascular endothelial cells (EC), and on translocation of phosphatidylserine (PS) to RBC surface, known as a potent mediator of RBC/EC adhesion, facilitating thrombus formation. Treatment of RBC with VitE strongly induces (up to sevenfold) PS externalization and enhances (up to 20-fold) their adherence to EC. The VitE hydrophilic analogue-Trolox-does not incorporate into cell membranes. Trolox did not exhibit any of these effects, implying that the VitE effect is due to its known ability to incorporate into cell membranes. The membrane-incorporated VitE significantly reduced the level of reactive oxygen species in H(2)O(2)-treated RBC, demonstrating that VitE elevates RBC/EC adhesion despite acting as an anti-oxidant. This study demonstrates for the first time that contrary to the common view of VitE as a beneficial supplement, VitE may introduce a circulatory risk by inducing flow-disturbing RBC adherence to blood vessel wall and the pro-thrombotic PS exposure. PMID- 20020329 TI - Circulating cytokines as biomarkers of alcohol abuse and alcoholism. AB - There are currently no consistent objective biochemical markers of alcohol abuse and alcoholism. Development of reliable diagnostic biomarkers that permit accurate assessment of alcohol intake and patterns of drinking is of prime importance to treatment and research fields. Diagnostic biomarker development in other diseases has demonstrated the utility of both open, systems biology, screening for biomarkers and more rational focused efforts on specific biomolecules or families of biomolecules. Long-term alcohol consumption leads to altered inflammatory cell and adaptive immune responses with associated pathologies and increased incidence of infections. This has led researchers to focus attention on identifying cytokine biomarkers in models of alcohol abuse. Alcohol is known to alter cytokine levels in plasma and a variety of tissues including lung, liver, and very importantly brain. A number of cytokine biomarker candidates have been identified, including: tumor necrosis factor-alpha, interleukin (IL)-1-alpha, IL-1-beta, IL-6, IL-8, IL-12, and monocyte chemoattractant protein-1. This is an emerging and potentially exciting avenue of research in that circulating cytokines may contribute to diagnostic biomarker panels, and a combination of multiple biomarkers may significantly increase the sensitivity and specificity of the biochemical tests aiding reliable and accurate detection of excessive alcohol intake. PMID- 20020331 TI - Stem cell transplants at childbirth. AB - Autologous transplantation of stem cells is a natural phenomenon at birth in mammals via the umbilical cord. Here, we discuss that a delay in the cord clamping may increase stem cell supply to the baby, thereby allowing an innate stem cell therapy that can render acute benefits in the case of neonatal disease, as well as long-term benefits against age-related diseases. PMID- 20020332 TI - Letter to the editor: Trans-upper-sternal approach to the cervicothoracic junction. PMID- 20020333 TI - The efficacy of periarticular multimodal drug infiltration in total hip arthroplasty. AB - BACKGROUND: Patient-controlled analgesia is a widely used and effective method of controlling pain after THA. This method is associated with substantial undesirable side effects. Local infiltration has been introduced in an attempt to reduce opioid requirements postoperatively, but its ability to reduce pain without complications is still questioned. QUESTIONS/PURPOSES: We evaluated patient-controlled analgesia use, pain and satisfaction scores, complication rates, and ropivacaine levels associated with the use of periarticular multimodal drug infiltration in THA. PATIENTS AND METHODS: We randomized 64 patients undergoing THA to receive a periarticular intraoperative multimodal drug injection or to receive no injection. All patients received patient-controlled analgesia for 24 hours after surgery. The final assessment was at 6 weeks. RESULTS: Patients receiving the periarticular injection used less patient controlled analgesia 6 hours postoperatively. The 24-hour patient-controlled analgesia requirement postsurgery also was less. The visual analog scale score for pain on activity in the postanesthetic care unit was less for patients who received an injection. The visual analog scale satisfaction score was similar in the two groups throughout the followup period. Recorded unbound ropivacaine levels were 2.5 times lower than toxic levels. CONCLUSIONS: Periarticular intraoperative injection with multimodal drugs can reduce postoperative patient controlled analgesia requirements and pain on activity in patients undergoing THA with no apparent increase in risk. LEVEL OF EVIDENCE: Level I, therapeutic study. See the guidelines online for a complete description of level of evidence. PMID- 20020334 TI - Case reports: subtrochanteric femoral stress fractures after prolonged alendronate therapy. AB - BACKGROUND: Alendronate is known for its ability to reduce bone loss in osteoporotic and osseous metastatic conditions. Its long-term effects remain unclear although several reports describe cases of proximal femur stress fractures associated with long-term alendronate use. CASE DESCRIPTION: We report the cases of four women who sustained low-energy subtrochanteric or femoral shaft stress fractures while being on alendronate therapy for more than 5 years. All radiographs showed typical patterns consisting of a transverse fracture line with external cortical bone reaction and medial cortical spike. Alendronate discontinuation along with nonoperative treatment was sufficient for one patient, whereas surgical stabilization was required in three patients. LITERATURE REVIEW: The side effects of alendronate therapy include osteonecrosis of the jaw, esophageal irritation, and musculoskeletal pain. Several cases of insufficiency femoral fractures associated with prolonged alendronate use have been reported. Their radiographic pattern and clinical presentation are consistent with our observations. Although various hypotheses have been suggested, the physiopathogenesis of these stress fractures is not completely understood. PURPOSES AND CLINICAL RELEVANCE: Although bisphosphonates play an important role in preventing pathologic fractures in patients with cancer, these subtrochanteric stress fractures associated with prolonged use of alendronate should not be ignored. PMID- 20020335 TI - Immunologic adverse reaction associated with low-carbide metal-on-metal bearings in total hip arthroplasty. AB - BACKGROUND: An increased incidence of periprosthetic osteolysis, resulting in loss of biologic fixation, has been reported in contemporary THAs with low carbide metal-on-metal compared with metal-on-polyethylene couple bearings. Although a hypersensitivity reaction attributable to Co and Cr debris is reportedly a potential cause for failure of THAs with high-carbide bearings, there are no evidence-based data for this reaction in low-carbide metal-on-metal bearings, although such hypersensitivity might be related to osteolysis. QUESTIONS/PURPOSES: We investigated whether there were differences in immunologic hypersensitivity reactions in retrievals from revised THAs with ceramic-on polyethylene versus metal-on-metal bearing couples. PATIENTS AND METHODS: We compared newly formed capsule and periprosthetic interface membranes from revision surgery for aseptic failure from 20 patients with low-carbide bearings and 13 patients with ceramic-on-polyethylene bearings. For control tissue, we obtained samples from the hip capsule during the primary THA implantation in 13 patients with low-carbide bearings and seven with ceramic-on-polyethylene bearings. We examined the tissues with conventional histologic and immunohistochemical methods. RESULTS: Compared with tissue from the control subjects and patients with ceramic-on-polyethylene bearings, the tissues from patients with low-carbide metal-on-metal bearings were associated with (1) extensive necrosis and fibrin exudation in the newly formed hip capsule and (2) diffuse and perivascular lymphocytic infiltration of a higher degree than in the hips with ceramic-on-polyethylene bearings in conventional histologic examination, and (3) more T than B cells. CONCLUSIONS: The conventional histologic and immunohistochemical findings in tissues retrieved from failed THAs with low-carbide metal-on-metal bearings are consistent with a link between hypersensitivity and osteolysis with low-carbide bearing couples. PMID- 20020336 TI - Proximal tibia osteoarticular allografts in tumor limb salvage surgery. AB - BACKGROUND: Resection of large tumors of the proximal tibia may be reconstructed with endoprostheses or allografts with fixation. Endoprosthetic replacement is associated with high failure rates and complications. Proximal tibia osteoarticular allografts after tumor resection allows restoration of bone stock and reconstruction of the extensor mechanism, but the long-term failure rates and complications are not known. QUESTIONS/PURPOSES: We therefore determined (1) the middle- and long-term survival of proximal tibia osteoarticular allografts, (2) their complications, and (3) functional (Musculoskeletal Tumor Society score) and radiographic (International Society of Limb Salvage) outcomes in patients treated with this reconstruction. PATIENTS AND METHODS: We retrospectively reviewed 52 patients (58 reconstructions including six repeat reconstructions) who underwent osteoarticular proximal tibia allograft reconstructions after resection of a bone tumor. The minimum followup of the 46 surviving patients was 72 months (mean, 123 months; range, 10-250 months). Survival of the allograft was estimated using the Kaplan-Meier method. We documented outcomes using the Musculoskeletal Tumor Society functional scoring system and the International Society of Limb Salvage radiographic scoring system. RESULTS: Six patients died from tumor-related causes without allograft failure before the 5-year radiographic followup. At last followup, 32 of the 52 remaining allografts were still in place; 20 failed owing to infections, local recurrences, or fractures. Overall allograft survival was 65% at 5 and 10 years, with an average Musculoskeletal Tumor Society functional score of 26 points and an average radiographic result of 87%. CONCLUSIONS: Based on these data we believe proximal tibia osteoarticular allograft is a valuable reconstructive procedure for large defects after resection of bone tumors. LEVEL OF EVIDENCE: Level IV, therapeutic study. See the Guidelines for Authors for a complete description of levels of evidence. PMID- 20020337 TI - Cost analysis of outpatient anterior cruciate ligament reconstruction: autograft versus allograft. AB - BACKGROUND: Prior studies suggest the cost of allograft anterior cruciate ligament (ACL) reconstruction is less than that for autograft reconstruction. Charges in these studies were influenced by patients requiring inpatient hospitalization. QUESTION/PURPOSE: We therefore determined if allograft ACL reconstruction would still be less costly if all procedures were performed in a completely outpatient setting. METHODS: We retrospectively reviewed 155 patients who underwent ACL reconstruction in an ambulatory surgery center between 2001 and 2004; 105 had an autograft and 50 had an allograft. Charges were extracted from itemized billing records, standardized to eliminate cost increases, and categorized for comparison. Surgeon and anesthesiologist fees were not included in the analysis. Groups were compared for age, gender, mean total cost, mean cost of implants, and several other cost categories. RESULTS: The mean total cost was $5465 for allograft ACL reconstruction and $4872 for autograft ACL reconstruction. There were no differences in complications between the two groups. CONCLUSIONS: Allograft ACL reconstruction was more costly than autograft ACL reconstruction in the outpatient setting. The cost of the allograft outweighs the increased surgical time needed for harvesting an autograft. LEVEL OF EVIDENCE: Level II, economic and decision analyses. See Guidelines for Authors for a complete description of levels of evidence. PMID- 20020338 TI - Differential effects of Tityus bahiensis scorpion venom on tetrodotoxin-sensitive and tetrodotoxin-resistant sodium currents. AB - We examined modification of sodium channel gating by Tityus bahiensis scorpion venom (TbScV), and compared effects on native tetrodotoxin-sensitive and tetrodotoxin-resistant sodium currents from rat dorsal root ganglion neurons and cardiac myocytes. In neurons, TbScV dramatically reduced the rate of sodium current inactivation, increased current amplitude, and caused a negative shift in the voltage-dependence of activation and inactivation of tetrodotoxin-sensitive channels. Enhanced activation of modified sodium channels was independent of a depolarizing prepulse. We identified two components of neuronal tetrodotoxin resistant current with biophysical properties similar to those described for NaV1.8 and NaV1.9. In contrast to its effects on neuronal tetrodotoxin-sensitive current, TbScV caused a small decrease in neuronal tetrodotoxin-resistant sodium current amplitude and the gating modifications described above were absent. A third tetrodotoxin-resistant current, NaV1.5 recorded in rat cardiac ventricular myocytes, was inhibited approximately 50% by TbScV, and the remaining current exhibited markedly slowed activation and inactivation. In conclusion, TbScV has very different effects on different sodium channel isoforms. Among the neuronal types, currents resistant to tetrodotoxin are also resistant to gating modification by TbScV. The cardiac tetrodotoxin-resistant current has complex sensitivity that includes both inhibition of current amplitude and slowing of activation and inactivation. PMID- 20020339 TI - Effects of stimuli intensity and frequency on auditory p50 and n100 sensory gating. AB - Sensory gating is the brain's ability to adjust its sensitivity to incoming stimuli, i.e., to diminish its response to irrelevant or repetitive stimuli (gating out) and to increase it when a novel stimulus is presented (gating in). Most of the existing studies have investigated the gating out mechanism, giving little attention to the gating in function. Although both the P50 and N100 components of the auditory ERPs (event related potentials) show amplitude reductions to stimuli repetition, it is not clear if both components are part of a common gating system or if their sensory modulation is uncorrelated. In order to respond to these questions and to further characterize the sensory gating functions, we examined to what extent P50 and N100 are influenced by changes in the stimuli parameters and whether the sensory modulation of both components are interrelated. To this end, we obtained ERPs from 23 healthy volunteers using pairs of auditory stimuli which could be identical (S1 = S2), different in frequency (S1 = 1000 Hz; S2 = 2000 Hz) or different in intensity (S1 = 80 dB SPL; S2 = 100 dB SPL). As expected, the amplitudes of P50 and N100 decreased in response to the second stimuli of the identical pairs. With non-identical pairs, amplitude increases of P50 and N100 were observed only in pairs with different intensity, but not frequency. Thus, the results showed that both P50 and N100 are sensory modulated, showing that amplitude decreased to stimuli repetition (gating out) and increased when the two stimuli of a pair differed in intensity (gating in). A correlational analysis of the sensory gating indices (S2/S1 ratio and S1 S2 difference) obtained for P50 and N100 suggested that the sensory gating function of both components may be of a different nature. The reliability of the ratio and the difference indices of sensory gating is also discussed. PMID- 20020340 TI - On building a memory evolutive system for application to learning and cognition modeling. AB - We address here aspects of the implementation of a memory evolutive system (MES), based on the model proposed by A. Ehresmann and J. Vanbremeersch (2007), by means of a simulated network of spiking neurons with time dependent plasticity. We point out the advantages and challenges of applying category theory for the representation of cognition, by using the MES architecture. Then we discuss the issues concerning the minimum requirements that an artificial neural network (ANN) should fulfill in order that it would be capable of expressing the categories and mappings between them, underlying the MES. We conclude that a pulsed ANN based on Izhikevich's formal neuron with STDP (spike time-dependent plasticity) has sufficient dynamical properties to achieve these requirements, provided it can cope with the topological requirements. Finally, we present some perspectives of future research concerning the proposed ANN topology. PMID- 20020341 TI - Agent-based cognitive model for human resources competence management. AB - This chapter presents an agent-based cognitive model aimed to represent human competency concepts and competence management processes of psychological nature. This model is implemented by a multiagent system application intended to help managers of software development projects to select, based on the competence management model, the right professionals to integrate a development team. There are several software engineering methodologies that can be used to design and develop multiagent systems. However, due to the necessity to handle human competency concepts of cognitive nature, like aptitudes, interests, abilities and knowledge, we were driven to choose methodologies that can handle these concepts since the inception of the system. To do so, we integrated the TROPOS methodology, and a set of software engineering methods derived from intelligent tutoring systems research to successfully analyze and design the proposed system. At the end of the paper we present a study case, showing how the proposed system should be applied to the domain of website development. PMID- 20020342 TI - Neural accumulator models of decision making in eye movements. AB - Humans and animals are constantly facing the problem of having to choose from a variety of possible actions as they interact with the environment. Both external and internal cues have to be used to guide their selection of a single action from many possible alternatives. Which action to choose in a given context may have important biological consequences to their survival. Decision making is regarded as an accumulation process of evidence about the state of the world and the utility of possible outcomes. Two well established neural accumulator models of decision making are presented to model the neural basis of decision making in behavioural paradigms such as the antisaccade task. PMID- 20020343 TI - On building meaning: a biologically-inspired experiment on symbol-based communication. AB - The use of an appropriate set of empirical and theoretical constraints to guide the construction of synthetic experiments leads to a better understanding of the natural phenomena under study, and allows for a greater understanding of the experimental results. We begin this chapter with a description of a general approach for conducting experiments with artificial creatures within a synthetic ethological context. Next, we describe how this approach was used to build a computational experiment regarding the emergence of self-organized symbols. Our experiment simulated a community of artificial creatures undergoing complex intra and inter-specific interactions in which meaning evolved over time, from a tabula rasa repertoire of random alarm-calls to a specific set of optimal referential alarm-calls. To design different kinds of creatures as well as innanimate elements of the environment, we applied theoretical constraints from the Peircean philosophy of sign and empirical constraints from neuroethology. Our results suggest that the constraints chosen were both necessary and sufficient to produce symbolic communication. PMID- 20020344 TI - Perception-action learning as an epistemologically-consistent model for self updating cognitive representation. AB - As well as having the ability to formulate models of the world capable of experimental falsification, it is evident that human cognitive capability embraces some degree of representational plasticity, having the scope (at least in infancy) to modify the primitives in terms of which the world is delineated. We hence employ the term 'cognitive bootstrapping' to refer to the autonomous updating of an embodied agent's perceptual framework in response to the perceived requirements of the environment in such a way as to retain the ability to refine the environment model in a consistent fashion across perceptual changes.We will thus argue that the concept of cognitive bootstrapping is epistemically ill founded unless there exists an a priori percept/motor interrelation capable of maintaining an empirical distinction between the various possibilities of perceptual categorization and the inherent uncertainties of environment modeling.As an instantiation of this idea, we shall specify a very general, logically-inductive model of perception-action learning capable of compact re parameterization of the percept space. In consequence of the a priori percept/action coupling, the novel perceptual state transitions so generated always exist in bijective correlation with a set of novel action states, giving rise to the required empirical validation criterion for perceptual inferences. Environmental description is correspondingly accomplished in terms of progressively higher-level affordance conjectures which are likewise validated by exploratory action.Application of this mechanism within simulated perception action environments indicates that, as well as significantly reducing the size and specificity of the a priori perceptual parameter-space, the method can significantly reduce the number of iterations required for accurate convergence of the world-model. It does so by virtue of the active learning characteristics implicit in the notion of cognitive bootstrapping. PMID- 20020345 TI - Detection of auditory cortex activity by fMRI using a dependent component analysis. AB - Functional MRI (fMRI) data often have low signal-to-noise-ratio (SNR) and are contaminated by strong interference from other physiological sources. A promising tool for extracting signals, even under low SNR conditions, is blind source separation (BSS), or independent component analysis (ICA). BSS is based on the assumption that the detected signals are a mixture of a number of independent source signals that are linearly combined via an unknown mixing matrix. BSS seeks to determine the mixing matrix to recover the source signals based on principles of statistical independence. In most cases, extraction of all sources is unnecessary; instead, a priori information can be applied to extract only the signal of interest. Herein we propose an algorithm based on a variation of ICA, called Dependent Component Analysis (DCA), where the signal of interest is extracted using a time delay obtained from an autocorrelation analysis. We applied such method to inspect functional Magnetic Resonance Imaging (fMRI) data, aiming to find the hemodynamic response that follows neuronal activation from an auditory stimulation, in human subjects. The method localized a significant signal modulation in cortical regions corresponding to the primary auditory cortex. The results obtained by DCA were also compared to those of the General Linear Model (GLM), which is the most widely used method to analyze fMRI datasets. PMID- 20020346 TI - Brain-computer interface using wavelet transformation and naive bayes classifier. AB - The main purpose of this work is to establish an exploratory approach using electroencephalographic (EEG) signal, analyzing the patterns in the time frequency plane. This work also aims to optimize the EEG signal analysis through the improvement of classifiers and, eventually, of the BCI performance. In this paper a novel exploratory approach for data mining of EEG signal based on continuous wavelet transformation (CWT) and wavelet coherence (WC) statistical analysis is introduced and applied. The CWT allows the representation of time frequency patterns of the signal's information content by WC qualiatative analysis. Results suggest that the proposed methodology is capable of identifying regions in time-frequency spectrum during the specified task of BCI. Furthermore, an example of a region is identified, and the patterns are classified using a Naive Bayes Classifier (NBC). This innovative characteristic of the process justifies the feasibility of the proposed approach to other data mining applications. It can open new physiologic researches in this field and on non stationary time series analysis. PMID- 20020347 TI - Neuromorphic systems: past, present and future. AB - Neuromorphic systems are implementations in silicon of elements of neural systems. The idea of electronic implementation is not new, but modern microelectronics has provided opportunities for producing systems for both sensing and neural modelling that can be mass produced straightforwardly. We review the the history of neuromorphic systems, and discuss the range of neuromorphic systems that have been developed. We discuss recent ideas for overcoming some of the problems, particularly providing effective adaptive synapses in large numbers. PMID- 20020348 TI - Genetic algorithm applied to hierarchically coupled associative memories. AB - Inspired by the theory of neuronal group selection (TNGS), we have carried out an analysis of the capacity of convergence of a multi-level associative memory based on coupled generalized-brain-state-in-a-box (GBSB) networks through evolutionary computation. The TNGS establishes that a memory process can be described as being organized functionally in hierarchical levels where higher levels coordinate sets of functions of lower levels. According to this theory, the most basic units in the cortical area of the brain are called neuronal groups or first-level blocks of memories and the higher-level memories are formed through selective strengthening or weakening of the synapses amongst the neuronal groups. In order to analyse this effect, we propose that the higher levels should emerge through a learning mechanism as correlations of lower level memories. According to this proposal, this paper describes a method of acquiring the inter-group synapses based on a genetic algorithm. Thus the results show that genetic algorithms are feasible as they allow the emergence of complex behaviours which could be potentially excluded in other learning process. PMID- 20020349 TI - Vector quantization of speech frames based on self-organizing maps. AB - We propose a speech compression technique based on vector quantization. A neural network with unsupervised learning is used to implement the vector quantizer. Some basic aspects related to speech signal processing are presented, as well as some general issues concerning the vector quantization problem. The idea of using a codebook to perform speech compression is introduced, and the use of a 2 dimensional self-organizing Kohonen map to generate the codebook is proposed. Simulation results are presented, giving some insights on the network topology, its initialization and training strategies, and codebook size. Finally, a comparison of speech quality obtained with our method and with a well-known compression algorithm is made. PMID- 20020350 TI - The use of bayesian networks for heart beat classification. AB - This work proposes to use a static Bayesian network as a tool to support medical decision in the on-line detection of Premature Ventricular Contraction beats (PVC) in electrocardiogram (ECG) records, which is a well known cardiac arrhythmia available for study in standard ECG databases. The main motivation to use Bayesian networks is their capability to deal with the uncertainty embedded in the problem (the medical reasoning itself frequently embeds some uncertainty). Indeed, the probabilistic inference is quite suitable to model this kind of problem, for considering its random character; as a consequence, random variables are used to propagate the uncertainty embedded in the problem. Some topologies of static Bayesian networks are implemented and tested in this work, in order to find out the one more suitable to the problem addressed. The results of such tests are discussed in details along the text, and the conclusions are highlighted. PMID- 20020351 TI - A histogram based method for multiclass classification using SVMs. AB - SVMs were primarily proposed to deal with binary classification. In this work an alternative O(log(2)(n)) method for multiple classes classification using SVMs is proposed. Experimental results showed that it can be 23 times faster than the one vs one method, and 1.3 times faster than the one vs all classic methods, with the same error rate. Tests were performed on a speaker independent, isolated word speech recognition scenario. PMID- 20020352 TI - A functional approach to emotion in autonomous systems. AB - The construction of fully effective systems seems to pass through the proper exploitation of goal-centric self-evaluative capabilities that let the system teleologically self-manage. Emotions seem to provide this kind of functionality to biological systems and hence the interest in emotion for function sustainment in artificial systems performing in changing and uncertain environments; far beyond the media hullabaloo of displaying human-like emotion-laden faces in robots. This chapter provides a brief analysis of the scientific theories of emotion and presents an engineering approach for developing technology for robust autonomy by implementing functionality inspired in that of biological emotions. PMID- 20020353 TI - A robot architecture based on higher order perception loop. AB - The paper discusses the self-consciousness of a robot as based on higher order perceptions of the robot itself. In this sense, the first order perceptions of the robot are the immediate perceptions of the outer world of the robot, while higher order perceptions are the robot perceptions of its own inner world. The resulting architecture based on higher order perceptions has been implemented and tested in a project regarding a robotic touristic guide acting in the Botanical Garden of the University of Palermo. PMID- 20020354 TI - The consciousness circuit - an approach to the hard problem. AB - This paper is the continuation of a study about Consciousness as a resulting function between attention and luminosity, presented at the Neuroscience International Congress, which was held in the city of Natal, in Brazil, in 2006. There it was described how visual Consciousness is generated by the interaction of the cortical area activity through FFS (Feedforward Sweep Processes), RP (Recurrent Processes) and WSRP (Wide Spread Recurrent Processes), and its relationship with the luminosity that hits the eyes. We have applied the reciprocal interaction model, which says that the eye reacts to luminosity through the regulation of sleep-awake states through EEG wave synchronization; this, in turn, is regulated by the thalamic cortical neural activity from the brainstem monoaminergic and cholinergic nuclei.Such an understanding has led us to construct a consciousness model which can be represented by an orthogonal graph. Through this model, we can represent all states of human consciousness (emotional consciousness, unconsciousness states, dreaming states, awareness states, pre-consciousness states and others) making it possible, in theory, to construct a Consciousness parameter which yields to the understanding of consciousness state observation without a subjective approach to experience.We have also applied on this orthogonal graph the Quantum Orch OR Model. According to it, subjective, phenomenal conscious vision depends on quantum computation in microtubules where the quantum is the smallest quantity of radiant energy, in a scale in which matter and energy interact. This model helps us to build the vertical axe of the consciousness orthogonal graph, a cholinergic-aminergic scale, in which activation triggers the quantum mechanism relevant to the consciousness phenomenon. PMID- 20020355 TI - Computational consciousness: building a self-preserving organism. AB - Consciousness has been a subject of crescent interest among the neuroscience community. However, building machine models of it is quite challenging, as it involves many characteristics and properties of the human brain which are poorly defined or are very abstract. Here I propose to use information theory (IT) to give a mathematical framework to understand consciousness. For this reason, I used the term "computational". This work is grounded on some recent results on the use of IT to understand how the cortex codes information, where redundancy reduction plays a fundamental role. Basically, I propose a system, here called "organism", whose strategy is to extract the maximal amount of information from the environment in order to survive. To highlight the proposed framework, I show a simple organism composed of a single neuron which adapts itself to the outside dynamics by taking into account its internal state, whose perception is understood here to be related to "feelings". PMID- 20020356 TI - The hippocampal system as the cortical resource manager: a model connecting psychology, anatomy and physiology. AB - A model is described in which the hippocampal system functions as resource manager for the neocortex. This model is developed from an architectural concept for the brain as a whole within which the receptive fields of neocortical columns can gradually expand but with some limited exceptions tend not to contract. The definition process for receptive fields is constrained so that they overlap as little as possible, and change as little as possible, but at least a minimum number of columns detect their fields within every sensory input state. Below this minimum, the receptive fields of some columns are expanded slightly until the minimum level is reached. The columns in which this expansion occurs are selected by a competitive process in the hippocampal system that identifies those in which only a relatively small expansion is required, and sends signals to those columns that trigger the expansion. These expansions in receptive fields are the information record that forms the declarative memory of the input state. Episodic memory activates a set of columns in which receptive fields expanded simultaneously at some point in the past, and the hippocampal system is therefore the appropriate source for information guiding access to such memories. Semantic memory associates columns that are often active (with or without expansions in receptive fields) simultaneously. Initially, the hippocampus can guide access to such memories on the basis of initial information recording, but to avoid corruption of the information needed for ongoing resource management, access control shifts to other parts of the neocortex. The roles of the mammillary bodies, amygdala and anterior thalamic nucleus can be understood as modulating information recording in accordance with various behavioral priorities. During sleep, provisional physical connectivity is created that supports receptive field expansions in the subsequent wake period, but previously created memories are not affected. This model matches a wide range of neuropsychological observation better than alternative hippocampal models. The information mechanisms required by the model are consistent with known brain anatomy and neuron physiology. PMID- 20020357 TI - Cognitive measure on different profiles. AB - Based on neurology and cognitive science many studies are developed to understand the human model mental, getting to know how human cognition works, especially about learning processes that involve complex contents and spatial-logical reasoning. Event Related Potential - ERP - is a basic and non-invasive method of electrophysiological investigation. It can be used to assess aspects of human cognitive processing by changing the rhythm of the frequency bands brain indicate that some type of processing or neuronal behavior. This paper focuses on ERP technique to help understand cognitive pathway in subjects from different areas of knowledge when they are exposed to an external visual stimulus. In the experiment we used 2D and 3D visual stimulus in the same picture. The signals were captured using 10 (ten) Electroencephalogram - EEG - channel system developed for this project and interfaced in a ADC (Analogical Digital System) board with LabVIEW system - National Instruments. That research was performed using project of experiments technique - DOE. The signal processing were done (math and statistical techniques) showing the relationship between cognitive pathway by groups and intergroups. PMID- 20020358 TI - History of doping and doping control. PMID- 20020359 TI - Biochemical and physiological aspects of endogenous androgens. AB - This review attempts to give a synopsis of the major aspects concerning the biochemistry of endogenous androgens, supplemented with several facets of physiology, particularly with respect to testosterone. Testosterone continues to be the most common adverse finding declared by World Anti-Doping Agency accredited laboratories, such samples having an augmented testosterone to epitestosterone ratio. Knowledge regarding the precursors and metabolism of endogenous testosterone is therefore fundamental to understanding many of the issues concerning doping with testosterone and its prohormones, including the detection of their administration. Further, adverse findings for nandrolone are frequent, but this steroid and 19-norandrostenedione are also produced endogenously, an appealing hypothesis being that they are minor by-products of the aromatization of androgens. At sports tribunals pertaining to adverse analytical findings of natural androgen administration, experts often raise issues that concern some aspect of steroid biochemistry and physiology. Salient topics included within this review are the origins and interconversion of endogenous androgens, the biosynthesis of testosterone and epitestosterone, the mechanism of aromatization, the molecular biology of the androgen receptor, the hypothalamic-pituitary-testicular axis, disturbances to this axis by anabolic steroid administration, the transport (binding) of androgens in blood, and briefly the metabolism and excretion of androgens. PMID- 20020360 TI - Phase-II metabolism of androgens and its relevance for doping control analysis. AB - Phase-II metabolism has a major contribution to androgen metabolism, converting the highly non-polar compounds to a more easily excreted form prior to their excretion in urine. In the human body the main phase-II metabolic reactions are glucuronidation and sulphonation. These reactions are catalysed by enzymes, which are categorised into families and further subfamilies based on their function and similarities of their amino-acid sequences. Due to inter-individual variation of the metabolising enzymes and their activities, the metabolic patterns of prohibited substances should be estimated for efficient doping control. In addition to target analytes the phase-II reactions have an effect on the selection of sample preparation procedure, chromatographic technique and ionisation method of the analysis routine. For method development and identification purposes adequate reference material is required, and to replace the laborious in vivo excretion studies, in vitro methodologies have been implemented to produce intact phase-II metabolites of androgens. PMID- 20020361 TI - Detecting the administration of endogenous anabolic androgenic steroids. AB - The detection of the administration of an androgen such as testosterone that could be present normally in human bodily fluids is based upon the methodical evaluation of key parameters of the urinary profile of steroids, precisely measured by GC/MS. Over the years, the markers of utilization were identified, the reference ranges of diagnostic metabolites and ratios were established in volunteers and in populations of athletes, and their stability in individual subjects was studied. The direct confirmation comes from the measurement of delta (13)C values reflecting their synthetic origin, ruling out a potential physiological anomaly. Several factors may alter the individual GC/MS steroid profile besides the administration of a testosterone-related steroid, the nonexhaustive list ranging from the microbial degradation of the specimen, the utilization of inhibitors of 5alpha-reductase or other anabolic steroids, masking agents such as probenecid, to inebriating alcohol drinking. The limitation of the testing strategy comes from the potentially elevated rate of false negatives, since only the values exceeding those of the reference populations are picked up by the GC/MS screening analyses performed by the laboratories on blind samples, excluding individual particularities and subtle doping. Since the ranges of normal values are often described from samples collected in Western countries, extrapolating data to all athletes appears inefficient. Furthermore, with short half-life and topical formulations, the alterations of the steroid profile are less pronounced and disappear rapidly. GC/C/IRMS analyses are too delicate and fastidious to be considered for screening routine samples. An approach based upon the individual athlete's steroid profiling is necessary to pick up variations that would trigger further IRMS analysis and investigations. PMID- 20020362 TI - Synthetic anabolic agents: steroids and nonsteroidal selective androgen receptor modulators. AB - The central role of testosterone in the development of male characteristics, as well as its beneficial effects on physical performance and muscle growth, has led to the search for synthetic alternatives with improved pharmacological profiles. Hundreds of steroidal analogs have been prepared with a superior oral bioavailability, which should also possess reduced undesirable effects. However, only a few entered the pharmaceutical market due to severe toxicological incidences that were mainly attributed to the lack of tissue selectivity. Prominent representatives of anabolic-androgenic steroids (AAS) are for instance methyltestosterone, metandienone and stanozolol, which are discussed as model compounds with regard to general pharmacological aspects of synthetic AAS. Recently, nonsteroidal alternatives to AAS have been developed that selectively activate the androgen receptor in either muscle tissue or bones. These so-called selective androgen receptor modulators (SARMs) are currently undergoing late clinical trials (IIb) and will be prohibited by the World Anti-Doping Agency from January 2008. Their entirely synthetic structures are barely related to steroids, but particular functional groups allow for the tissue-selective activation or inhibition of androgen receptors and, thus, the stimulation of muscle growth without the risk of severe undesirable effects commonly observed in steroid replacement therapies. Hence, these compounds possess a high potential for misuse in sports and will be the subject of future doping control assays. PMID- 20020363 TI - Nandrolone: a multi-faceted doping agent. AB - Nandrolone or nortestosterone, an anabolic-androgenic steroid, has been prohibited by doping control regulations for more than 30 years. Although its main metabolism in the human body was already known at that time, and detection of its misuse by gas or liquid chromatographic separation with mass spectrometric detection is straightforward, many interesting aspects regarding this doping agent have appeared since.Over the years, nandrolone preparations have kept their position among the prohibited substances that are most frequently detected in WADA-accredited laboratories. Their forms of application range from injectable fatty acid esters to orally administered nandrolone prohormones. The long detection window for nandrolone ester preparations and the appearance of orally available nandrolone precursors have changed the pattern of misuse.At the same time, more refined analytical methods with lowered detection limits led to new insights into the pharmacology of nandrolone and revelation of its natural production in the body.Possible contamination of nutritional supplements with nandrolone precursors, interference of nandrolone metabolism by other drugs and rarely occurring critical changes during storage of urine samples have to be taken into consideration when interpreting an analytical finding.A set of strict identification criteria, including a threshold limit, is applied to judge correctly an analytical finding of nandrolone metabolites. The possible influence of interfering drugs, urine storage or natural production is taken into account by applying appropriate rules and regulations. PMID- 20020364 TI - Designer steroids. AB - Anabolic steroids have been studied for over 50 years and during that time numerous compounds with a variety of functional groups have been produced and many have been published. Of these only a small number have been introduced to the pharmaceutical market. WADA has continued the work begun by the IOC banning the use of these agents within sport as performance enhancing substances. Athletes, however, continue to use these anabolic steroids but tighter testing and the introduction of unannounced sample collection has made this form of cheating harder.In order to try to evade detection, athletes who continue to dope are having to resort to the use of a far more dangerous form of drug - the designer steroid. These steroids are manufactured to closely resemble existing known compounds, but with sufficient chemical diversity to ensure that their detection by the WADA accredited laboratories is more difficult. A worrying feature of the use of these compounds is that no data is available to evaluate either the efficacy or the safety of these substances. Many such drugs are now being made in clandestine ways (as demonstrated by the recent BALCO case) and then passed on to athletes who become the guinea pigs determining the potential of the substances as doping agents.Methods for the detection of these new compounds are being developed using emerging techniques such as gas chromatography or liquid chromatography attached to a variety of mass spectrometry instruments. This technology as well as vigilance by laboratories and enforcement agencies can all help in early detection of designer steroids being used for doping. PMID- 20020365 TI - Growth hormone. AB - Human growth hormone (hGH) is a proteohormone secreted by the pituitary gland. It acts through binding to the hGH receptor, inducing either direct effects or initiating the production of insulin-like growth-factor I (IGF-I), the most important mediator of hGH effects. Growth hormone is primarily known to promote longitudinal growth in children and adolescents, but has also various important metabolic functions throughout adult life. Effects of hGH on the adult organism are well established from studies with recombinant growth hormone (rhGH) therapy in growth hormone deficient subjects. In this particular group of patients, replacement of hGH leads to increased lipolysis and lean body mass, decreased fat mass, improvements in VO(2max), and maximal power output. Although extrapolation from these findings to the situation in well trained healthy subjects is impossible, and controlled studies in healthy subjects are scarce, abuse of hGH seems to be popular among athletes trying to enhance physical performance. Detection of the application of rhGH is difficult, especially because the amino acid sequence of rhGH is identical to the major 22,000 Da isoform of hGH normally secreted by the pituitary. Furthermore, some physiological properties of hGH secretion also hindered the development of a doping test: secreted in a pulsatile manner, it has a very short half-life in circulation, which leads to highly variable serum levels. Concentration alone therefore cannot prove the exogenous administration of hGH.Two approaches have independently been developed for the detection of hGH doping: The so-called "marker approach" investigates changes in hGH-dependent parameters like IGF-I or components of bone and collagen metabolism, which are increased after hGH injection. In contrast, the so-called "isoform approach" directly analyses the spectrum of molecular isoforms in circulation: the pituitary gland secretes a spectrum of homo- and heterodimers and - multimers of a variable spectrum of hGH isoforms, whereas rhGH consists of the monomeric 22,000 Da isoform only. This isoform therefore becomes predominant after injection of rhGH. Specific immunoassays with preference for the one or the other isoform allow analysis of the relative abundance of the 22,000 Da isoform. Application of rhGH can be proven when the ratio of this isoform relative to the others is increased above a certain threshold. Because the "marker method" and the "isoform method" have a different window of opportunity for detection, complementary use of both tests could be a way to increase the likelihood of detecting cheating athletes. PMID- 20020366 TI - Mass spectrometry-based analysis of IGF-1 and hGH. AB - Mass spectrometric approaches have been used to determine various peptide hormones in sports drug testing. While insulin-like growth factor-1 (IGF-1) and its synthetic analogs are qualitatively and/or quantitatively measured by liquid chromatography-tandem mass spectrometry after immunoaffinity purification, methods of uncovering doping rule violations with illegal applications of human growth hormone (hGH) have not yet been established using mass spectrometry-based assays. However, substantial information on the heterogeneity of hGH, splice variants and post-translational modifications with respective locations as elucidated by mass spectrometry are of utmost importance for improving currently employed immunological procedures. PMID- 20020367 TI - Insulin. AB - Due to its versatile nature and its corresponding anabolic and anticatabolic properties, insulin has been prohibited in sports since 1999. Numerous studies concerning its impact on glycogen formation, protein biosynthesis, and inhibition of protein breakdown have illustrated its importance for healthy humans and diabetics as well as elite athletes. Various reports described the misuse of insulin to improve performance and muscle strength, and synthetic analogs were the subject of several studies describing the beneficial effects of biotechnologically modified insulins. Rapid- or long-acting insulins were developed to enhance the injection-to-onset profile as well as the controllability of administered insulin, where the slightest alterations in primary amino acid sequences allowed the inhibition of noncovalent aggregation of insulin monomers (rapid-acting analogs) or promoted microprecipitation of insulin variants upon subcutaneous application (long-acting analogs). Information on the metabolic fate and renal elimination of insulins has been rather limited, and detection assays for doping control purposes were primarily established using the intact compounds as target analytes in plasma and urine specimens. However, recent studies revealed the presence of urinary metabolites that have been implemented in confirmation methods of sports drug testing procedures. So far, no screening tool is available providing fast and reliable information on possible insulin misuse; only sophisticated procedures including immunoaffinity purification followed by liquid chromatography and tandem mass spectrometry have enabled the unambiguous detection of synthetic insulins in doping control blood or urine samples. PMID- 20020368 TI - beta-Adrenergic stimulation. AB - Two groups of substances which stimulate the adrenergic system are listed as prohibited by the World Anti-Doping Agency. Stimulants are prohibited in competition only and beta(2)-agonists are prohibited in- and out-of-competition. While beta(2)-agonists act directly on the target receptors, sympathomimetic amines can exert their action directly and indirectly. Due to differences in pharmacology but mainly due to differences in administered dose, differences in detection methods between both groups of substances exist although preparation is similar and consists of an extraction at basic pH. Gas chromatography coupled to mass spectrometry has been the detection methodology of choice for several decades. However, the importance of liquid chromatography-mass spectrometry as a preferred detection methodology is rapidly increasing, especially for the detection of beta(2)-agonists and new additions to the list of prohibited stimulants, such as modafinil. Pharmacology, metabolism and detection of both groups of prohibited substances will be discussed. PMID- 20020369 TI - Erythropoietin and analogs. AB - Erythropoietin (EPO), a glycoprotein hormone, stimulates the growth of red blood cells and as a consequence it increases tissue oxygenation. This performance enhancing effect is responsible for the ban of erythropioetin in sports since 1990. Especially its recombinant synthesis led to the abuse of this hormone, predominatly in endurance sports. The analytical differentiation of endogenously produced erythropoietin from its recombinant counterpart by using isoelectric focusing and double blotting is a milestone in the detection of doping with recombinant erythropoietin. However, various analogous of the initial recombinant products, not always easily detectable by the standard IEF-method, necessitate the development of analytical alternatives for the detection of EPO doping. The following chapter summarizes its mode of action, the various forms of recombinant erythropoietin, the main analytical procedures and strategies for the detection of EPO doping as well as a typical case report. PMID- 20020370 TI - Blood transfusion in sports. AB - Blood transfusion is an effective and unmediated means of increasing the number of red blood cells in the circulation in order to enhance athletic performance. Blood transfusion became popular in the 1970s among elite endurance athletes and declined at the end of the 1980s with the introduction of recombinant erythropoietin. The successive implementation in 2001 of a direct test to detect exogenous erythropoietin and in 2004 of a test to detect allogeneic blood transfusion forced cheating athletes to reinfuse fully immunologically compatible blood. The implementation of indirect markers of blood doping stored in an Athlete's Biological Passport provides a powerful means to deter any form of blood transfusion. PMID- 20020371 TI - The athlete's biological passport and indirect markers of blood doping. AB - In the fight against doping, disciplinary sanctions have up to now been primarily based on the discovery of an exogenous substance in a biological fluid of the athlete. However, indirect markers of altered erythropoiesis can provide enough evidence to differentiate between natural variations and blood doping. Forensic techniques for the evaluation of the evidence, and more particularly Bayesian networks, allow antidoping authorities to take into account firstly the natural variations of indirect markers - through a mathematical formalism based on probabilities - and secondly the complexity due to the multiplicity of causes and confounding effects - through a distributed and flexible graphical representation. The information stored in an athlete's biological passport may be then sufficient to launch a disciplinary procedure against the athlete. The strength of the passport is that it relies on a statistical approach based on sound empirical testing on large populations and justifiable protocols. Interestingly, its introduction coincides with the paradigm shift that is materializing today in forensic identification science, from archaic assumptions of absolute certainty and perfection to a more defensible empirical and probabilistic foundation. PMID- 20020372 TI - Masking and manipulation. AB - The list of prohibited substances in sports includes a group of masking agents that are forbidden in both in- and out-of-competition doping tests. This group consists of a series of compounds that are misused in sports to mask the administration of other doping agents, and includes: diuretics, used to reduce the concentration in urine of other doping agents either by increasing the urine volume or by reducing the excretion of basic doping agents by increasing the urinary pH; probenecid, used to reduce the concentration in urine of acidic compounds, such as glucuronoconjugates of some doping agents; 5alpha-reductase inhibitors, used to reduce the formation of 5alpha-reduced metabolites of anabolic androgenic steroids; plasma expanders, used to maintain the plasma volume after misuse of erythropoietin or red blood cells concentrates; and epitestosterone, used to mask the detection of the administration of testosterone. Diuretics may be also misused to achieve acute weight loss before competition in sports with weight categories. In this chapter, pharmacological modes of action, intended pharmacological effects for doping purposes, main routes of biotransformation and analytical procedures used for anti-doping controls to screen and confirm these substances will be reviewed and discussed. PMID- 20020373 TI - Hormonal growth promoting agents in food producing animals. AB - In contrast to the use of hormonal doping agents in sports to enhance the performance of athletes, in the livestock industry hormonal growth promoters ("anabolics") are used to increase the production of muscle meat. This leads to international disputes about the safety of meat originating from animals treated with such anabolics.As a consequence of the total ban in the EU of all hormonal active growth promoters ("hormones") in livestock production, in contrast to their legal use [e.g. of five such hormones (17beta-estradiol, testosterone, progesterone, trenbolone and zeranol) as small solid ear implants and two hormones as feed additives for feedlot heifers (melengestrol acetate) and for swine (ractopamine) in the USA], the regulatory controls also differ sharply between the EU and the USA.In the EU the treatment of slaughter animals is the regulatory offence that has to be controlled in inspection programs. In the USA testing for compliance of a regulatory maximum residue level in the edible product (muscle, fat, liver or kidney) is the purpose of the inspection program (if any).The EU inspection programs focus on sample materials that are more suitable for testing for banned substances, especially if the animals are still on the farm, such as urine and feces or hair. In the case of slaughtered animals, the more favored sample materials are bile, blood, eyes and sometimes liver. Only in rare occasions is muscle meat sampled. This happens only in the case of import controls or in monitoring programs of meat sampled in butcher shops or supermarkets.As a result, data on hormone concentrations in muscle meat samples from the EU market are very rare and are obtained in most cases from small programs on an ad hoc basis. EU data for natural hormones in meat are even rarer because of the absence of "legal natural levels" for these hormones in compliance testing. With the exception of samples from the application sites - in the EU the site of injection of liquid hormone preparations or the site of application of "pour on" preparations - the hormone concentrations observed in meat samples of illegally treated animals are typically in the range of a few micrograms per kilogram (ppb) down to a few tenths of a microgram per kilogram. In the EU dozens of illegal hormones are used and the number of active compounds is still expanding. Besides estrogenic, androgenic and progestagenic compounds also thyreostatic, corticosteroidal and beta-adrenergic compounds are used alone or in "smart" combinations.An overview is given of the compounds identified on the EU black market. An estimate is also given of the probability of consumption in the EU of "highly" contaminated meat from the application sites in cattle. Finally some data are presented on the concentration of estradiol in bovine meat from animals treated and not treated with hormone implants. These data are compared with the recent findings for estradiol concentrations in hen's eggs. From this comparison, the preliminary conclusion is that hen's eggs are the major source of 17alpha- and 17beta-estradiol in the consumer's daily "normal" diet. PMID- 20020374 TI - Some aspects of doping and medication control in equine sports. AB - This chapter reviews drug and medication control in equestrian sports and addresses the rules of racing, the technological advances that have been made in drug detection and the importance of metabolism studies in the development of effective drug surveillance programmes. Typical approaches to screening and confirmatory analysis are discussed, as are the quality processes that underpin these procedures. The chapter also addresses four specific topics relevant to equestrian sports: substances controlled by threshold values, the approach adopted recently by European racing authorities to control some therapeutic substances, anabolic steroids in the horse and LC-MS analysis in drug testing in animal sports and metabolism studies. The purpose of discussing these specific topics is to emphasise the importance of research and development and collaboration to further global harmonisation and the development and support of international rules. PMID- 20020375 TI - Androgenic anabolic steroid abuse and the cardiovascular system. AB - Abuse of anabolic androgenic steroids (AAS) has been linked to a variety of different cardiovascular side effects. In case reports, acute myocardial infarction is the most common event presented, but other adverse cardiovascular effects such as left ventricular hypertrophy, reduced left ventricular function, arterial thrombosis, pulmonary embolism and several cases of sudden cardiac death have also been reported. However, to date there are no prospective, randomized, interventional studies on the long-term cardiovascular effects of abuse of AAS. In this review we have studied the relevant literature regarding several risk factors for cardiovascular disease where the effects of AAS have been scrutinized:(1) Echocardiographic studies show that supraphysiologic doses of AAS lead to both morphologic and functional changes of the heart. These include a tendency to produce myocardial hypertrophy (Fig. 3), a possible increase of heart chamber diameters, unequivocal alterations of diastolic function and ventricular relaxation, and most likely a subclinically compromised left ventricular contractile function. (2) AAS induce a mild, but transient increase of blood pressure. However, the clinical significance of this effect remains modest. (3) Furthermore, AAS confer an enhanced pro-thrombotic state, most prominently through an activation of platelet aggregability. The concomitant effects on the humoral coagulation cascade are more complex and include activation of both pro coagulatory and fibrinolytic pathways. (4) Users of AAS often demonstrate unfavorable measurements of vascular reactivity involving endothelial-dependent or endothelial-independent vasodilatation. A degree of reversibility seems to be consistent, though. (5) There is a comprehensive body of evidence documenting that AAS induce various alterations of lipid metabolism. The most prominent changes are concomitant elevations of LDL and decreases of HDL, effects that increase the risk of coronary artery disease. And finally, (6) the use of AAS appears to confer an increased risk of life-threatening arrhythmia leading to sudden death, although the underlying mechanisms are still far from being elucidated. Taken together, various lines of evidence involving a variety of pathophysiologic mechanisms suggest an increased risk for cardiovascular disease in users of anabolic androgenic steroids. PMID- 20020376 TI - Side effects of anabolic androgenic steroids: pathological findings and structure activity relationships. AB - Side effects of anabolic steroids with relevance in forensic medicine are mainly due to life-threatening health risks with potential fatal outcome and cases of uncertain limitations of criminal liability after steroid administration. Both problems are typically associated with long-term abuse and excessive overdose of anabolic steroids. Side effects may be due to direct genomic or nongenomic activities (myotrophic, hepatotoxic), can result from down-regulation of endogenous biosynthesis (antiandrogenic) or be indirect consequence of steroid biotransformation (estrogenic).Logically, there are no systematic clinical studies available and the number of causally determined fatalities is fairly limited. The following compilation reviews typical abundant observations in cases where nonnatural deaths (mostly liver failure and sudden cardiac death) were concurrent with steroid abuse. Moreover, frequent associations between structural characteristics and typical side effects are summarized. PMID- 20020377 TI - Gene doping. AB - Gene doping abuses the legitimate approach of gene therapy. While gene therapy aims to correct genetic disorders by introducing a foreign gene to replace an existing faulty one or by manipulating existing gene(s) to achieve a therapeutic benefit, gene doping employs the same concepts to bestow performance advantages on athletes over their competitors. Recent developments in genetic engineering have contributed significantly to the progress of gene therapy research and currently numerous clinical trials are underway. Some athletes and their staff are probably watching this progress closely. Any gene that plays a role in muscle development, oxygen delivery to tissues, neuromuscular coordination, or even pain control is considered a candidate for gene dopers. Unfortunately, detecting gene doping is technically very difficult because the transgenic proteins expressed by the introduced genes are similar to their endogenous counterparts. Researchers today are racing the clock because assuring the continued integrity of sports competition depends on their ability to develop effective detection strategies in preparation for the 2012 Olympics, which may mark the appearance of genetically modified athletes. PMID- 20020378 TI - Science and the rules governing anti-doping violations. AB - The fight against the use of performance-enhancing drugs in sports has been in effect for nearly 90 years. The formation of the World Anti-Doping Agency in 1999 was a major event because an independent agency was entrusted with harmonization of the antidoping program. In addition to sports governing bodies, governments have endorsed WADA and its programs by signing a United Nations Education, Science, and Cultural Organization Convention on Doping. The first step in the harmonization process was the development of the World Anti-Doping Program. This program consisted of five documents - the Code, the International Standard for Testing, the International Standard for Laboratories, the Prohibited List, and the International Standard for Therapeutic Use Exemptions - which unified the approach of the international federations and national antidoping agencies in applying antidoping rules. For laboratory testing, the International Standard for Laboratories establishes the performance expectations for and competence of laboratories recognized by WADA, including accreditation under ISO/IEC 17025. The antidoping rules are adjudicated by arbitration using the internationally recognized Court of Arbitration for Sport. PMID- 20020379 TI - [Topical examples for a broad and dynamic development of paediatric surgery]. PMID- 20020380 TI - [Fetoscopic surgery as prenatal intervention for isolated congenital diaphragmatic hernia]. AB - Congenital diaphragmatic hernia (CDH) occurs sporadically with an incidence of 1:2,500 live births. Despite the progress in neonatal intensive care, CDH remains associated with a mortality of at least 30 % in isolated cases. The in essence surgically correctable defect of the diaphragm enables the prenatal herniation of abdominal organs into the thoracic cavity. The resulting abnormal development of the airways and pulmonary vessels causes neonatal respiratory insufficiency and persistent pulmonary hypertension. The condition can be diagnosed prenatally and the degree of pulmonary hypoplasia, which determines the postnatal course, can be measured to make an -individual prognosis. In severely affected patients, prenatal surgery may improve neonatal outcome by reversing pulmonary hypoplasia. This is currently implemented by percutaneous fetoscopic endoluminal tracheal occlusion (FETO) to trigger fetal lung growth. Although there are no maternal complications, preterm rupture of the membranes remains the major drawback of the procedure (20 % < 34 weeks). However, as compared to historical controls of a similar severity, survival as well as early neonatal morbidity are significantly improved by FETO. As a consequence, a multicentre randomised-controlled trial in fetuses with moderate hypoplasia on FETO compared to expectant management has been started ( www.totaltrial.eu). Primary outcome measure is survival without chronic lung disease (i. e., with-out bronchopulmonary dysplasia). A trial in severely affected -fetuses with survival as main outcome is currently under review by ethics committee. A standardised neonatal management enables optimal treatment and multicentre compatibility. It remains to be proven if fetoscopic surgery can maintain a solid position in the prenatal treatment of CDH to improve both mortality and morbidity of the affected children. PMID- 20020381 TI - [Functional outcome in children with Hirschsprung's disease or imperforate anus]. AB - Various outcomes following operative therapy for Hirschsprung's disease and anorectal malformations have been reported. Operative techniques for anorectal reconstruction have been modified several times in the past. Repair of anorectal malformations have been performed through a posterior sagittal approach since the 1980s. This -allows an anatomically correct reconstruction of the anorectal canal. Abdominoperineal or sacro-abdominoperineal pull-through procedures, as the classical operative techniques, have been abandoned by most surgeons. Rectosigmoid-ectomy with colo-anal anastomosis, as described by Swenson and Bill in 1948, as well as the retro-rectal pull-through (Duhamel) and the endorectal pull-through (Soave) are still frequently used in surgery for Hirschsprung's disease. The development of the transanal endorectal pull-through with (Georgeson) or without (de la Torre) laparoscopic assistance has eliminated the necessity of laparotomy in selected cases. Despite significant progress in the understanding of the pathophysiology of and therapy for Hirschsprung's dis-ease and anorectal malformations, the functional results remain unsatisfactory. Functional problems occur already in early childhood and de-crease the quality of life significantly. Although complications resolve with time and the quality of life normalises in adolescence and adulthood, this might be mainly due to an adaptation strategy by the patients. For the future, a standardised and prospective study design is necessary to compare different procedures and to provide a basis for the further development of therapeutic strategies. PMID- 20020382 TI - [Fast-track paediatric surgery]. AB - Fast-track surgery is a comprehensive perioperative treatment concept that has been successfully performed and widely accepted in adult surgery since the 1990s. The crucial aim is to speed up convalescence and to avoid perioperative complications as pneumonia and thrombosis. Compared to conventional treatment strategies, hospital stays are substantially reduced. In the paediatric field fast-track surgery is not generally established. However, in recent studies a high efficiency of paediatric surgical fast-track procedures with respect to medical, psychological, economical and ethical parameters has been shown. It has been confirmed that early convalescence leads to an increase of satisfaction of the patients and their parents without higher complication rates. Shorter hospital stays lead to reduced expenses for the health insurances and parents. Fast-track concepts are not implemented in the German reimbursement system G-DRG. Thus, problems with intensified nursing and reimbursement remain to be solved. PMID- 20020383 TI - [Thoracic surgery in neonates and infants with congenital malformations]. AB - Congenital malformations of the lung and diaphragm are a challenge in paediatric surgery. Depending on the malformation they show a broad spectrum of symptoms and a varying age at manifestation. Thus there are many diagnostic and therapeutic options, which require a good knowledge of the pathology. The treatment of these complex cases should lead to early referral to paediatric centres containing an interdisciplinary team with neonatologists, paediatric pulmonologists and cardiologists, ENT surgeons, anaesthesiologists, radiologists and paediatric surgeons. Some malformations are diagnosed prenatally and need intrauterine interventions. Decisive is the early diagnosis and treatment of these malformations. Nowadays the surgical therapy of neonates and infants with malformations of the lung and diaphragm is enriched by a number of endoscopic and endoluminal techniques, which are discussed critically in this article. PMID- 20020384 TI - [Congenital abdominal wall defects--an analysis of prevalence and operative management by means of gastroschisis and omphalocele]. AB - The management of congenital abdominal wall defects is one of the main characteristics of quality for a department of paediatric surgery. The results of treatment in the early years were the reason for a continuous improvement of procedures, operation strategies and the kinds of -material that had been used. During the last years there has been a great discussion about the presumed increase in the incidence of gastroschisis and the preterm delivery of such cases. On the basis of our own first results (2006-2009) and details from the literature, we survey these two topics. PMID- 20020385 TI - [Bariatric surgery in children and adolescents]. AB - Obesity in childhood and adolescents has gained epidemic proportions; in Germany 15-20 % of boys and girls are overweight, more than 6 % are known to be obese. By now, 25 % of relevant people show a pathological glucose intolerance, 4-5 % are developing type 2 diabetes mellitus (T2DM). In addition, metabolic disorders leading to hypertension and cardiac, renal or ophthalmological complications could be named as serious comorbidities. Medical and behavioural intervention as treatment for obesity in childhood remains largely ineffective: 5-10 % weight loss within 2 years rarely results in significant durable success. In adults, bariatric surgery is being used increasingly as an effective approach to achieve weight loss and to improve serious medical comorbidities, in particular T2DM. Enhancement of quality of life and explicit extension of survival are concomitant phenomenons. To date, a range of different types of bariatric procedures has been performed in adolescents, but studies evaluating and analysing preoperative data, postoperative course and follow-up in a representative number of patients younger than 18 years are still lacking. Nevertheless, current experience suggests significant weight loss and improving obesity-related medical comorbidities after bariatric surgery in adolescents too. Moreover, bariatric surgery in adolescents seems to induce less complications and a shorter hospital stay than in adults. Al though surgical therapy for obesity in this group of patients remains an individual decision, even though explicit guidelines have been published specifying inclusion and exclusion criterias. Analysis of our own patient group and results of the study of the quality assurance "surgical treatment of morbid obesity" are appropriate tools to evaluate surgical techniques and to provide long-term follow-up. PMID- 20020386 TI - [Principles of oncological surgery for lung metastases in paediatric solid tumours]. AB - Distant metastases regularly occur in cases of paediatric solid tumours. Overall the most common localisation of metastases is the lung. The correct treatment of lung metastases represents a prognostic factor in most paediatric solid tumours. Guidelines for the therapy of lung metastases are commonly included in the protocols of the different multicentric treatment trials. However, specific surgical aspects are usually not addressed in these protocols. The aim of this article is to present the principles of surgery for lung metastases in paediatric solid tumours. The principles of surgery for metastases of the most common extracranial paediatric solid tumours are presented including aspects of the diagnostic work-up, surgical preparations, and surgical procedures. Special points of interest are discussed focusing on the different entities. Surgery for lung metastases in paediatric solid tumours can be performed with a good surgical outcome regardless of the patients' age. Large numbers of sequelae can be surgically excised. Depending on the localisation sternotomy is an option concerning the surgical approach. Surgery for lung metastases in paediatric solid tumours is a safe method when respecting the principles for the procedure. It has a positive influence on the patients' prognosis. An aggressive surgical procedure may be justified depending on the tumour entity. An interdisciplinary approach including paediatric oncology and radiology is mandatory in any case. PMID- 20020387 TI - [Percutaneous laparoscopic hernia repair in children]. AB - Laparoscopic repair of inguinal hernia in children is not established in the clinical routine so far. Especially the higher rate of recurrences was the main reason to choose open conventional herniotomy. With the development of microinvasive percutaneous extraperitoneal closure, the recurrence rate decreased significantly and is now comparable to that after open hernia repair. Different clinical studies as well as our own experience have confirmed these outcomes. PMID- 20020388 TI - [Mechanical ileus in childhood--causes and surgical therapy]. AB - BACKGROUND: The present study examines the causes and surgical management in children who underwent surgery for a mechanical Ileus. PATIENTS: We studied all children up to the age of 18 years who had undergone a surgical intervention for a mechanical ileus between 1.1.1996 and 31.12.2006. 89 children were included in this retrospective study. RESULTS: Of the total of 89 children 15 were newborn (16.9 %), 23 babies (25.8 %), 19 toddlers (21.3 %) and 32 school children (36 %) at the time of the operation. 51 of the 89 children had undergone at least one previous abdominal operation. Intraoperative findings showed the cause for the ileus to be adhesions in 56 and a bowel invagination in the remaining 11 children. Associated malformations were found in 34 children, the most frequent being malformations of the heart and gastrointestinal tract. The most frequent surgical intervention was adhesiolysis in 56 children (62.9 %), followed by the reposition of invaginated intestine in 11 (12.4 %). Bowel resection was necessary in 23 children (25.8 %). CONCLUSION: The risk for developing an ileus due to adhesions increases with the number of previous operations. Surgical intervention for an ileus aims to decompress the overstretched bowel and to restore gastrointestinal flow by removing the mechanical obstruction. One third of the children with an ileus have accompanying malformations. Children with a mechanical ileus should undergo surgery as soon as possible. PMID- 20020389 TI - [Dislocated basicervical fracture of the femoral neck in a 7-year-old girl]. AB - This report discusses the treatment of a seven-year-old girl with a displaced fracture of the proximal femur (AO 32-M / 3.1 II, Delbet type III) after falling 4 m from a tree. She was treated by immediate (within 6 h after trauma) open reduction and capsular decompression. The fracture was stabilised with 2 cannulated titanium screws that did not go beyond the epiphyseal cartilage. Clinical and radiological examination after 2 years showed no complications, especially no sign of avascular necrosis (AVN) of the femoral head. The functional outcome with Ratliff's classification was good. In accordance with the latest literature, we recommend immediate open reduction with accurate reposition, capsular decompression, and stabile internal fixation in order to treat dislocated complete basicervical fractures. This combination minimises the devastating complications of AVN for these fractures and various other complications including malunion, non-union, leg-length discrepancy, premature epiphyseal closure, coxa vara and valga. PMID- 20020390 TI - [Incidental finding of an acute appendicitis in a premature newborn with haematochezia]. AB - In newborns, acute appendicitis is a very rare condition associated with significant lethality. Due to mostly non-specific symptoms, it is difficult to find the correct diagnosis preoperatively. Interestingly, rectal bleeding as a clinical sign in neonatal appendicitis is very uncommon. Here, we report on a 4 day-old premature female newborn with rectal bleeding who, therefore, underwent laparotomy because of a suspected volvulus. Except for an acutely inflamed appendix, no other pathological findings were found intraoperatively, leading to appendectomy. Histological investigation of the specimen confirmed acute ulcero phlegmonous appendicitis. Thus, the rectal bleeding can be attributed to erosions as part of the inflammatory changes in clinically apparent appendicitis. The postoperative course of the patient was unremarkable, in perticular, no further rectal bleeding episode was observed. In spite of the low incidence of neonatal appendicitis, it has to be included in the spectrum of differential diagnoses if unclear abdominal discomfort occurs and whenever non-specific clinical signs are found in newborns. Early surgical intervention is considered the curative treatment approach of choice and can, thus, contribute to a reduction of the potential complications. PMID- 20020391 TI - [De Garengeot appendicitis--rare variant of a common medical condition]. AB - De Garengeot hernia is an extremely rare pathological finding. Here, we report the case of a 69-year-old woman who presented with acute appendicitis within a femoral hernia. Although exceptionally rare, the clinician needs to be aware of this variation in order to avoid complications arising due to delays in diagnosis and therapy. PMID- 20020392 TI - [Duodenal perforation after blunt abdominal trauma]. AB - Duodenal perforation after a blunt abdominal trauma is a rare emergency situation that can result in life-threatening complications. We report on a woman who had a perforation of the duodenum after a supposed mild blunt abdominal trauma. Unremarkable at the initial presentation, the patient presented with acute abdominal pain and a retroperitoneal abscess five days after the initial trauma. The duodenal repair was performed with a Roux-Y anastomosis. Difficulties in diagnosis are very common, but the early recognition of the rupture is essential. The contrast-enhanced CT scan is the gold standard for diagnosis. Surgical management depends on the severity of the trauma and must be chosen on an individual basis. PMID- 20020393 TI - [Gallbladder duplication--laparoscopic cholecystectomy 17 years after open cholecystectomy]. AB - INTRODUCTION: Duplication of the gallbladder is a rare congenital anomaly. An incidence of 2.5:10,000 has been published in autoptic studies. METHODS: We carried out an analysis of the published case reports of the last 30 years to examine those cases in which an accessory gallbladder was pre- or intraoperatively detected or missed and to evaluate the implications of the time of diagnosis of the duplication on the surgical therapy. RESULTS: 28 case reports were analysed. If the duplication of the gallbladder was recognised pre operatively or during operation, both gallbladders could be removed via laparoscopy in 80 % of the cases. Missing the second gallbladder can lead to persisting symptoms, postoperative complications as well as a recrudescence of the cholecystolithiasis with following reoperation. If a reoperation for a missed second gallbladder was necessary, only 14.3 % of these operations could be performed via laparoscopy. CASE REPORT: We report the case of a missed gall bladder duplication with an acute cholecystitis causing a reoperation 17 years after the initial operation. A laparoscopic cholecystectomy was possible even though the initial operation was performed through open surgery. CONCLUSION: If recognised in preoperative examinations or during surgery a laparoscopic cholecystectomy of both gallbladders is possible in the majority of cases with duplicated gallbladder. PMID- 20020394 TI - Long-term follow-up of neuroblastoma-associated opsoclonus-myoclonus-ataxia syndrome. AB - OBJECTIVE: The aim of this study is to describe the long-term neurological, neuropsychological and neuroradiological sequelae and to determine prognostic factors for neurological outcome in children with neuroblastoma-associated opsoclonus-myoclonus-ataxia (OMA) syndrome. METHODS: Data on medical history were collected for the study patients. Examinations with grading of neurological signs, neuropsychological tests and brain magnetic resonance imaging with spectroscopy were performed during a follow-up clinic. RESULTS: Fourteen subjects entered the study. All had localized neuroblastoma and they were evaluated after a median of 7.8 years. Patients with a chronic/multiphasic neurological course received steroids combined with intravenous immunoglobulins in the majority of cases. 71% presented neurological sequelae and 62% had a full-scale IQ below the normal range. All patients showed at least some deficit in the neuropsychological functions assessed (language, visual-motor integration, memory, attention and motor ability). Long-term deficits were more frequently detected in patients with an interval of more than 2 months between OMA onset and its diagnosis, even if in most comparisons statistical significance was not reached. Cerebellar atrophy, observed in 36% of patients, was not associated with the neurological outcome. CONCLUSIONS: Persisting disability is present in most children with neuroblastoma associated OMA. However, our results support the role of an early diagnosis of OMA in reducing sequelae and encourage the use of new immunosuppressive therapies. PMID- 20020395 TI - Outcome of extremely premature infants at early school age: health-related quality of life and neurosensory, cognitive, and behavioral outcomes in a population-based sample in northern Germany. AB - PURPOSE: The study aimed at collecting regional data to support and establish evidence-based decision-making. METHODS: We investigated a cohort of 154 preterm infants with gestational age <27+0 weeks born between 1997 and 1999 in a defined region of Northern Germany regarding neurosensory and cognitive outcomes, overall disability status, behavioral problems, and health-related quality of life at the age of seven to nine years (mean: eight years, SD seven months). RESULTS: 92 (60%) infants survived, 75 of these 92 (82%) were followed-up. Rates of disability were high: only 27 (36%) showed 'no dysfunction' of neurosensory status, 33 (44%) 'mild dysfunction', 5 (7%) 'moderate dysfunction' and 10 (13%) 'severe dysfunction', including 8 (11%) with non-ambulatory cerebral palsy. 19 (26%) were mentally retarded. Parents reported behavioral problems in 21 (28%), health-related quality of life was lower in preterm infants compared to a representative normal sample. In multivariate analyses IVH III-IV/PVL was an independent risk factor for adverse outcomes. Behavior problems were predicted by low IQ and lower educational level of the mother. CONCLUSIONS: Overall our results confirm high levels of mortality and morbidity in extremely immature infants. Regional data should include mortality, morbidity and health-related quality of life to adequately inform parents about the prognosis. PMID- 20020396 TI - Epilepsy in patients with propionic acidemia. AB - Propionic acidemia (PA) is an autosomal recessively inherited defect of propionyl CoA carboxylase with an incidence of approximately 1:50 000. There are few reports on the occurrence of EEG findings and development of epilepsy in patients with PA. Retrospectively, the data of 17 patients with PA from one Italian and four Austrian centers were evaluated concerning EEG findings and the development of epilepsy. Nine patients showed a disturbance of background activity, as well as epileptiform discharges. All nine patients with pathological EEG discharges developed seizures compatible with the definition of symptomatic epilepsy. Five of these nine patients showed fever induced seizures at the beginning. Two of them suffered from symptomatic absence epilepsy. Six of the nine patients with seizures were treated with antiepileptic drugs (AED), which were tolerated without side-effects. Four patients showed photosensitivity, which so far has never been reported in PA. We hypothesize that patients with PA are prone to cortical dysfunction caused by one or several pathological metabolites - leading to changes in background and epileptiform activity with a high manifestation rate of clinical seizures. PMID- 20020397 TI - Neurological outcome and risk of recurrence depending on the anterior vs. posterior arterial distribution in children with stroke. AB - We report the outcome of 46 previously healthy children with arterial ischemic stroke. After a mean follow-up of 26 months, five (11%) children suffered a recurrence and 28 (61%) were left with sequelae. The prevalence and the severity of the sequelae were similar irrespective of whether the localization of the accident was anterior or posterior. However, a recurrence was significantly more frequent in the posterior than in the anterior group (4/14 vs. 1/32; p=0.025). These observations may lead to the establishment of therapeutic guidelines according to the localization of the infarct. PMID- 20020398 TI - Andermann syndrome can be a phenocopy of hereditary motor and sensory neuropathy- report of a discordant sibship with a compound heterozygous mutation of the KCC3 gene. AB - Andermann syndrome is a rare autosomal recessive disorder characterized by agenesis of the corpus callosum (ACC), progressive motor-sensory neuropathy, mental retardation and facial features. We report on two siblings with the clinical picture of a demyelinating hereditary motor and sensory neuropathy (HMSN), where only the presence of ACC in the younger brother pointed to the diagnosis of Andermann syndrome. Mutation analysis of the KCC3 (SLC12A6) gene showed a compound heterozygous mutation; a maternal missense mutation c.1616G>A (p.G539D) and a paternal splice mutation c.1118+1G>A in both siblings. We hypothesize that mutations of the KCC3 gene may result in non-syndromic childhood onset HMSN. PMID- 20020399 TI - Aplasia cutis congenita--plastic reconstruction of three scalp and skull defects with two opposed scalp rotation flaps and split thickness skin grafting. AB - Aplasia cutis congenita (ACC) is a rare congenital defect of skin and subcutaneous tissue, more rarely of periosteum, skull and dura. The lesions can involve any location, but most common are scalp defects. We report on the successful treatment of three large defects of the scalp with skull involvement in a newborn girl by early debridement and defect closure with two opposed scalp rotation flaps and an occipital split-thickness skin graft. PMID- 20020400 TI - Association of syndromic mental retardation and autism with 22q11.2 duplication. AB - We describe a 5.3-year-old girl with autism, mental retardation, hypotonia, marked speech delay, and mild dysmorphic features with a 22q11.2 duplication. Her mother carries the same duplication and presents cleft palate, and normal intelligence. The clinical and behavioural phenotype of this relatively new syndrome is very heterogeneous, with high variability also in the familiar cases. Up till now, about 50 cases of 22q11.2 duplication have been reported, but only three of them are associated with autistic disorders. We propose that in addition to 22q13.3 deletion syndrome, also 22q11.2 duplication should be suspected in a patient with unspecified dysmorphisms, mental retardation, autism, hypotonia, and severe speech delay. PMID- 20020401 TI - Congenital infiltrating lipomatosis of the face with hemimegalencephaly. AB - Congenital infiltrating lipomatosis of the face (CILF) is a rare disorder, causing unilateral facial asymmetry characterized by enlargement of the cheek or chin. Hemimegalencephaly is a unique malformation characterized by enlargement of a cerebral hemisphere. The association of CILF and hemimegalencephaly has rarely been reported. We present a case of unilateral facial swelling in a 1.5-year-old boy in whom magnetic resonance imaging and computed tomography revealed CILF associated with unilateral megalencephaly. PMID- 20020402 TI - Central pontine and extrapontine myelinolysis in a pediatric patient following rapid correction of hypernatremia. AB - Central pontine and extrapontine myelinolysis are uncommon disorders characterized by distinctive clinical features and typical findings on neuroimaging. Only a few cases are reported in the pediatric age group. We describe the case of a leukemic, malnourished 14-year-old boy with a high serum sodium concentration that gradually increased to 170 mmol/L. During a septic shock episode, hydration with a low sodium concentration at the rate of 104 mL/h for 24 h was administered. A rapid correction of the high serum sodium occurred, exceeding 0.5 mmol/L/h. The following day the patient developed rapid and progressive neurological impairment with clinical features characteristic of central pontine and extrapontine myelinolysis. Magnetic resonance imaging confirmed the diagnosis 11 days later. The patient was treated with steroids and intravenous immunoglobulins. He achieved an almost full neurological recovery and radiological improvement. The reported case demonstrates that central pontine and extrapontine myelinolysis can occur after excessively rapid correction of hypernatremia. PMID- 20020403 TI - Congenital and evolving vascular disorders associated with cutaneous hemangiomas: case report. AB - Conventional arteriography in an 11-month-old boy with cardiopathy, aortic arch coarctation and haemangiomas showed the absence of the right internal carotid and vertebral arteries, hypertrophy of the right external carotid artery, with enlargement of the internal maxillary and ophthalmic arteries that supplied the right cerebral hemisphere. An MRI study showed an infarcted area in the posterior zone of the left cerebral hemisphere vascularised by the middle cerebral artery that was caused by a thrombosis during a severe bout of gastroenteritis. MRA studies performed at 16 and 23 years of age revealed progressive narrowing of the left carotid and vertebral arteries, persistence of the proatlantal and trigeminal arteries, and poor cerebral vascularisation that, at adult age, was entirely supplied through collateral arteries, branches of both external carotids, the presence of unilateral duplication of the vertebral arteries and ascending pharyngeal artery. PMID- 20020405 TI - Epilepsy and respiratory chain defects in children. PMID- 20020406 TI - Visual attention deficits contribute to impaired facial emotion recognition in boys with attention deficit/hyperactivity disorder. PMID- 20020407 TI - Tissue engineering toward organ replacement: a promising approach in airway transplant. AB - Autologous tissue transfer, allografts and prosthetic replacements have so far failed to offer functional solutions for the treatment of long circumferential tracheal defects. Because of the shortcomings related with these strategies, interest has turned increasingly to the field of tissue engineering which applies the principles of engineering and life sciences in an effort to develop in vitro biological substitutes able to restore, maintain, or improve tissue and organ function. The advances in this field during the past decade have thus provided a new attractive approach toward the concept of functional substitutes and may represent an alternative to the shortage of suitable grafts for reconstructive airway surgery. This article gives an overview of the tissue engineering approach and of the encouraging strategies attempted so far in trachea regeneration. PMID- 20020408 TI - Alginate-encapsulated human hepatoma C3A cells for use in a bioartificial liver device - the hybrid-MDS. AB - PURPOSE: The aim of this study was to encapsulate C3A cells into alginate microcapsules with an average diameter of < or =100 microm, thus enabling them to be recirculated in a bioartificial liver device based on MDS (Microsphere-based Detoxification System) technology. The microcapsules have to be permeable for essential proteins such as albumin. METHODS: C3A cells were encapsulated using alginate. The resulting alginate beads were coated with poly(diallyldimethylammoniumchloride) (pDADMAC) and poly(sodium-p styrenesulfonate) (pSS). Their mechanical stability was tested by recirculation of the microcapsule suspension, while their permeability was determined by reverse-size exclusion chromatography and by the use of a confocal laser microscope. The metabolic activities of encapsulated C3A cells were compared to freely growing adherent C3A cells in static cultivation models. The metabolic functionality of encapsulated C3A cells in static conditions was compared to encapsulated C3A cells in a dynamic model. RESULTS: The mean diameter of the resulting microcapsules was 86 mum. Our experiments show that these microcapsules were permeable for albumin and the high flow rate of 600 ml/min in a dynamic model has no influence on the survival and the metabolic activities of the encapsulated cells during the tested time of 24 hours. CONCLUSIONS: Alginate microcapsules containing C3A cells can be used to produce albumin and growth factors in a bioartificial or hybrid liver support system. Thanks to their small diameter, the microcapsules in suspension can be recirculated in the MDS. PMID- 20020409 TI - Comparison of fluid status in patients treated by different modalities of peritoneal dialysis using multi-frequency bioimpedance. AB - BACKGROUND AND AIMS: Patients treated by peritoneal dialysis, especially those using automatic cyclers, are frequently found to be hypervolemic. To determine whether there are significant differences between the currently available modalities of peritoneal dialysis, we analyzed whether differences in transporter status and treatment modality had a discernible effect on extracellular fluid volumes as measured by multi-frequency bioimpedance. METHODS: Two hundred prevalent peritoneal dialysis patients, 48% male, mean age 54.9 yr (SD+/-15.6), were studied using multi-frequency bioimpedance following a standard peritoneal equilibration test; 63 patients were treated by CAPD, 29 by APD, 96 by CCPD and 12 by OCCPD. RESULTS: There were no differences in extracellular volumes, or extracellular volume adjusted for height, or as a ratio to total body water between the groups. As expected, extracellular volume adjusted for height depended upon bodyweight (r=0.412, p<0.001), sex (r=0.457, p<0.001) and systolic blood pressure (r=0.162, p=0.023), and extracellular volume to total body water related to the reciprocal of total daily peritoneal ultrafiltration losses (r= 0.0254, p=0.003) and urine output (-0.254, p=0.003). More importantly, on logistical regression analysis the ratio of extracellular fluid to total body fluid increased with falling albumin, F=21.5 p<0.001, increasing age, F=18.5 p<0.001, urine output F=6.46, p=0.014, total daily ultrafiltration, F=3.52 and protein intake p=0.003. Extracellular fluid adjusted for total body fluid was associated with CRP (males F=6.03, p=0.03, females F=4.438, p=0.04). CONCLUSION: Patients were more likely to have an expanded extracellular fluid volume if they had reduced daily fluid losses, but also with biomarkers typically associated with poor nutrition and inflammation. PMID- 20020410 TI - Hemoglobin variability in patients with chronic kidney disease in the Netherlands. AB - PURPOSE: Hemoglobin cycling has been reported in hemodialysis patients treated with erythropoiesis-stimulating agents (ESA) and is associated with increased mortality. Information on hemoglobin cycling in Europe is limited. We investigated hemoglobin variability in the Netherlands. Hemodialysis and peritoneal dialysis patients were studied and pre-dialysis patients were enrolled. METHODS: This observational retrospective study was executed in a Dutch dialysis center. We studied 157 patients from 2005 to 2007: 56 hemodialysis, 12 peritoneal dialysis and 29 pre-dialysis patients, all treated with ESA; and 60 pre-dialysis patients without ESA. Patients were divided on the basis of their pattern of hemoglobin fluctuation around a range of 11-12 g/dL. In dialysis patients, the amount of time that hemoglobin remained within that range was calculated. For all patients, the magnitude of hemoglobin fluctuations was assessed (i.e. the difference between hemoglobin maximum and minimum) and data on ESA dose changes and hospitalizations were collected. RESULTS: None of the ESA treated patients had hemoglobin levels stable within the target range over a one year period. Pre-dialysis patients without ESA also showed variable hemoglobin levels. A stepwise decrease in the magnitudes of hemoglobin fluctuation was observed in the hemodialysis patients, peritoneal dialysis patients, pre-dialysis patients using ESA, and the pre-dialysis patients without ESA, respectively. CONCLUSION: In the Netherlands, hemoglobin variability is common in hemodialysis and peritoneal dialysis patients, but also in pre-dialysis patients. The results of this study warrant further research into the relationship between hemoglobin variability and clinical outcomes. PMID- 20020411 TI - Development of a new combined test setup for accelerated dynamic pH-controlled in vitro calcification of porcine heart valves. AB - Fifty years after their first implantation, bioprosthetic heart valves still suffer from tissue rupture and calcification. Since new bioprostheses exhibit a lower risk of calcification, fast and reliable in vitro methods need to be evaluated for testing the application of new anti-calcification techniques. This report describes a modification of the well-known in vitro dynamic calcification test method (Glasmacher et al, Leibniz University Hannover (LUH)), combined with the pH-controlled, constant solution supersaturation (CSS) method (University of Patras (UP)). The CSS method is based on monitoring the pH of the solution and the addition of calcium and phosphate ion solutions through the implementation of two syringe pumps. The pH and the activities of all ions in the solutions are thus kept constant, resulting in higher calcification rates compared to conventional in vitro methods in which solution supersaturation is allowed to decrease without any further control. To verify this hypothesis, five glutaraldehyde preserved porcine aortic valves were tested. Three of the valves were tested according to a free-drift methodology: the valves were immersed in a supersaturated calcification solution, with an initial total calcium times total phosphate product of (CaxP)=10.5 (mmol/L)2, renewed weekly. Two valves were tested by the new pH-controlled loop system, implementing the CSS methodology. All valves were tested for a 4-week period, loaded at 300 cycles per minute, resulting in a total of 12 million cycles at the end of the testing period. The degree of calcification was determined weekly by means of mux-ray, and by conventional, clinical and micro-computer tomography (CT, muCT). The results showed that the valves mineralizing at constant solution supersaturation in vitro yielded higher rates of calcification compared to the valves tested at conditions of decreasing solution supersaturation without any control, indicating the development of a new, accelerated, controllable in vitro calcification method. PMID- 20020412 TI - Clinical and biomaterial evaluation of a new condensed dual-function extracorporeal circuit in reoperation for coronary artery bypass surgery. AB - PURPOSE: This prospective, randomized study compared the clinical performance of three types of circuits: a newly introduced, fully-coated, interchangeable open closed circuit with a dual configuration (hard shell with a bypass shunt), reduced length, and reduced prime of less than 800 cc (CondECC); a completely coated circuit (ECC); and a similar uncoated, open circuit with standard length and prime (CONT). METHODS: 75 patients undergoing reoperation for coronary revascularization were randomly allocated into three groups (N=25): Group 1: CondECC with shortened tubing, components and an open-closed configuration of low priming volume with a centrifugal pump and a shunt which bypassed the reservoir for closed configuration; Group 2: ECC with a roller pump and hard-shell reservoir; Group 3: CONT. Blood samples for CBC, inflammatory mediators [Interleukin-2 (IL-2), Complement-3a (C3a)] and flow cytometry (CD11b/CD18) were collected after induction (T1) and heparin administration (T2), 15 min after cardiopulmonary bypass (CPB) (T3), before cessation of CPB (T4), 15 min after reversal (T5), and the first postoperative day (T6). RESULTS: Leukocyte counts demonstrated significant increases at T4, T5 in CONT but remained stable in ECC and CondECC (p<0.05). Platelets were preserved better at T4, T5 in both ECC and CondECC study groups (p<0.05). IL-2 and C3a levels were significantly lower at T3, T4, T5 in CondECC and T4, T5 in ECC (p<0.05). Blood protein adsorption analysis demonstrated increased amount of microalbumin on CONT fibers (p<0.05). CONCLUSIONS: The CondECC is a flexible, dual-function, open/closed configuration system that was easy to use, safe and achieved better biocompatibility when compared to coated and uncoated conventional circuits. PMID- 20020414 TI - Fully automated liquid extraction-based surface sampling and ionization using a chip-based robotic nanoelectrospray platform. AB - A fully automated liquid extraction-based surface sampling device utilizing an Advion NanoMate chip-based infusion nanoelectrospray ionization system is reported. Analyses were enabled for discrete spot sampling by using the Advanced User Interface of the current commercial control software. This software interface provided the parameter control necessary for the NanoMate robotic pipettor to both form and withdraw a liquid microjunction for sampling from a surface. The system was tested with three types of analytically important sample surface types, viz., spotted sample arrays on a MALDI plate, dried blood spots on paper, and whole-body thin tissue sections from drug dosed mice. The qualitative and quantitative data were consistent with previous studies employing other liquid extraction-based surface sampling techniques. PMID- 20020413 TI - Osteogenic differentiation of human mesenchymal stromal cells on surface-modified titanium alloys for orthopedic and dental implants. AB - PURPOSE: Surface properties of titanium alloys, used for orthopedic and dental applications, are known to affect implant interactions with host tissues. Osteointegration, bone growth and remodeling in the area surrounding the implants can be implemented by specific biomimetic treatments; these allow the preparation of micro/nanostructured titanium surfaces with a thickened oxide layer, doped with calcium and phosphorus ions. We have challenged these experimental titanium alloys with primary human bone marrow stromal cells to compare the osteogenic differentiation outcomes of the cells once they are seeded onto the modified surfaces, thus simulating a prosthetic device-biological interface of clinical relevance. METHODS: A specific anodic spark discharge was the biomimetic treatment of choice, providing experimental titanium disks treated with different alkali etching approaches. The disks, checked by electron microscopy and spectroscopy, were subsequently used as substrates for the proliferation and osteogenic differentiation of human cells. Expression of markers of the osteogenic lineage was assessed by means of qualitative and quantitative PCR, by cytochemistry, immunohistochemistry, Western blot and matrix metalloprotease activity analyses. RESULTS: Metal surfaces were initially less permissive for cell growth. Untreated control substrates were less efficient in sustaining mineralized matrix deposition upon osteogenic induction of the cells. Interestingly, bone sialo protein and matrix metalloprotease 2 levels were enhanced on experimental metals compared to control surfaces, particularly for titanium oxide coatings etched with KOH. DISCUSSION: As a whole, the KOH modification of titanium surfaces seems to allow the best osteogenic differentiation of human mesenchymal stromal cells, representing a possible plus for future clinical prosthetic applications. PMID- 20020415 TI - Inhibition of the p53-MDM2 interaction by adenovirus delivery of ribosomal protein L23 stabilizes p53 and induces cell cycle arrest and apoptosis in gastric cancer. AB - BACKGROUND: Inhibiting MDM2 activity in tumors that express wild-type (wt) p53 but have high levels of MDM2 protein has been considered an attractive anticancer strategy for many years. Previous studies revealed that human ribosomal protein L23 (RPL23) inhibited MDM2-mediated p53 degradation and thus induced p53 levels as well as its activity, suggesting that it might be a candidate for use as a gene therapy for cancer. In the present study, we evaluated whether targeting this pathway could be of therapeutic value against human gastric carcinoma. METHODS: Gastric cancer cell lines carrying wt or mutant p53 gene were infected with adenovirus expressing RPL23 (Ad-RPL23). Cell growth assay, flow cytometry assay and morphology were used to observe the effects of Ad-RPL23 infection on tumor cells, and further, the effect of Ad-RPL23 treatment on tumor growth in vivo was investigated. RESULTS: In vitro, adenovirus-mediated RPL23 gene transfer stabilized wt p53 by inhibiting its degradation, and thus resulted in G(1)-S cell cycle arrest and/or apoptosis of human gastric cancer MKN45 and AGS cells carrying wt p53 gene. Adenovirus-mediated RPL23 gene transfer also inhibited the growth of MKN45 tumors in subcutaneous mouse models. CONCLUSIONS: The data obtained in the present study suggest that, through the inhibition of the p53 MDM2 interaction, adenovirus delivery of RPL23 can inhibit the proliferation of gastric cancer cells harboring wt p53 in vitro and in vivo. Exogenous RPL23 induced wt p53 stabilization and activation may be a novel therapeutic approach for patients with gastric cancer. PMID- 20020416 TI - A sensitive and specific HPGPC-FD method for the study of pharmacokinetics and tissue distribution of Radix Ophiopogonis polysaccharide in rats. AB - Interest in antimyocardial ischemic activity of a graminan-type fructan with a weight average molecular weight of 4.8 kDa extracted from Radix Ophiopogonis (ROP) has necessitated the study of its pharmacokinetics and tissue distribution. For that, a simple HPGPC-FD method was developed for the sensitive and specific determination of FITC-ROP (fluorescein-isothiocyanate-labeled ROP) in plasma and rat tissues (heart, liver, spleen, lung, kidney, brain and stomach). The analyte was separated on a Shodex Sugar KS-802 high-performance gel column with 0.1 M phosphate buffer (pH 7.0) as mobile phase at a flow rate of 0.5 mL/min, and fluorescence detection at lambda(ex) 495 nm and lambda(em) 515 nm. The calibration curve for FITC-ROP was linear over the range 0.25-20.0 or 50.0 microg/mL in all studied biosamples with correlation coefficients > 0.995. The inter-day and intra-day precisions of analysis were not more than 10%, and assay accuracy ranged from 93 to 105% for plasma and from 89 to 108% for tissue homogenates. This method has been confirmed here to be suitable for the study of pharmacokinetics and tissue distribution of ROP and the achieved results are highly instructive for the further pharmaceutical development of ROP, suggesting the promising application of the method to the increasingly important carbohydrate-based drugs. PMID- 20020417 TI - Determination of eflornithine enantiomers in plasma by precolumn derivatization with o-phthalaldehyde-N-acetyl-L-cysteine and liquid chromatography with UV detection. AB - A bioanalytical method for indirect determination of eflornithine enantiomers in 75 microL human plasma has been developed and validated. L- and D-eflornithine were derivatized with o-phthalaldehyde and N-acetyl-L-cysteine to generate diastereomers which were separated on two serially connected Chromolith Performance columns (RP-18e 100 x 4.6 mm i.d.) by a isocratic flow followed by a gradient flow for elution of endogenous compounds. The diastereomers were detected with UV (340 nm). The between-day precisions for L- and D-eflornithine in plasma were 8.4 and 2.3% at 3 microm, 4.0 and 5.1% at 400 microm, and 2.0 and 3.7% at 1000 microm. The lower limit of quantification was determined to be 1.5 microm, at which precision was 14.9 and 9.9% for L- and D-eflornithine, respectively. PMID- 20020418 TI - Metabolic stability and determination of cytochrome P450 isoenzymes' contribution to the metabolism of medetomidine in dog liver microsomes. AB - Medetomidine is a potent and selective alpha2-adrenergic agonist. The activation of alpha2-adrenergic receptor mediates a variety of effects including sedation, analgesia, relief of anxiety, vasoconstriction and bradycardia. However, our main interest is the sedative effects of medetomidine when used as a premedicant prior surgery in companion animals, especially in dogs. Recently, data suggested that following intravenous infusion at six dosing regiments non-linear pharmacokinetics was observed. Major causes of non-linear pharmacokinetics are the elimination of the drug not following a simple first-order kinetics and/or the elimination half-life changing due to saturation of an enzyme system. The goal of this study was to establish the metabolic stability and determine the metabolic pathway of medetomidine in dog liver microsomes. Consequently, Michaelis-Menten parameters (V(max), K(m)), T(1/2) and CL(i) were determined. The incubations were performed in a microcentrifuge tube and containing various concentrations of medetomidine (10-5000 nM), 1 mg/mL of microsomal proteins suspended in 0.1 M phosphate buffer, pH 7.4. Microsomal suspensions were preincubated with NADPH (1 mM) for 5 min at 37 degrees C prior to fortification with medetomidine. Samples were taken at various time points for kinetic information and the initial velocity (v(i)) was determined after 10 min incubation. The reaction was stopped by the addition of an internal standard solution (100 ng/mL of dextrometorphan in acetone). Medetomidine concentrations were determined using a selective and sensitive HPLC-ESI/MS/MS method. Using non linear regression, we determined a K(m) value of 577 nM, indicating relatively low threshold enzyme saturation consistent with previous in vivo observation. The metabolic stability was determined at a concentration of 100 nm (<1)abeo-taxane diterpenoids, taiwantaxins A-C (1-3, resp.) are lacking an O bearing functionality at either C(13) or C(14). Compound 2 showed significant cytotoxic activity against human PC-3 tumor cells. PMID- 20020458 TI - Variability in foliar essential oils among different morphotypes of Lantana species complexes, and its taxonomic and ecological significance. AB - The genus Lantana has many species complexes, and L. camara is one of the aggressive alien weedy species complexes; species delimitation in these complexes is a nightmare for taxonomists. We examined the diversity in the chemical composition of foliar essential oils among morphotypes of Lantana species complexes inhabiting the same ecological gradient, and its taxonomic and ecological significance. The yields of essential oils varied from 0.1 to 0.79% in foliar hydrodistillates of eleven morphotypes, and a total of 39 chemical constituents were detected by GC/MS. The quantitative and qualitative variability in the composition of essential oils among morphotypes was very high, and hence they represent chemotypes. The diversity observed in the composition of essential oils appears to be of genetic origin and thus of taxonomic value. The formation of distinct clusters and sub-clusters at high distance cluster combine values also substantiates that the patterns of distribution of chemical constituents among morphotypes can be used in delimiting species and infraspecific taxa within the species complexes. The presence of beta-caryophyllene and other such compounds, which are known to prevent herbivory, in morphotypes of Lantana species complexes suggest that these compounds may provide selective advantage to Lantana over native species in the invasion of new and disturbed habitats. PMID- 20020459 TI - Synthesis and biophysical studies of bis-macrocyclic cobalt/copper(II) complexes having a pyridine spacer with CT DNA and 5'-GMP. AB - New bis-macrocyclic complexes of Co(III), 1, Ni(II), 2, and Cu(II), 3, containing pyridyl bridges between 13-membered macrocyclic subunits, have been synthesized via an in situ one-pot template condensation reaction (IOPTCR). The proposed structures of these new dinuclear complexes are consistent with the data obtained from elemental analysis, molar conductance, IR, EPR, UV/VIS, (1)H- and 13C-NMR, and ESI-MS. The complexes 2 and 3 possess square-planar geometry with four secondary N-atoms coordinated to the metal ion, while complex 1 reveals octahedral geometry in solution due to coordinated H(2)O molecules. DNA-Binding properties of the complexes 1 and 3 were investigated by absorption and emission titrations, cyclic voltammetry, and viscosity measurements. Complexes 1 and 3 are strong DNA binders with binding constants, K(b), of 1.64 x 10(5) and 2.05 x 10(5) M(-1), respectively. Hyperchromism, decrease in emission intensity of DNA-bound ethidium bromide (EB), and changes observed in the viscosity and cyclic voltammograms in the presence of added metal complexes reveals that the complexes bind to DNA predominantly by electrostatic attraction, substantiated by absorption titration with 5'-GMP. PMID- 20020460 TI - Sinuladiterpenes A-F, new cembrane diterpenes from Sinularia flexibilis. AB - Chromatographic investigation of the octocoral Sinularia flexibilis afforded six new cembrane diterpenes, sinuladiterpenes A-F (1-6, resp.), in addition to four known cembranolides, 11-episinulariolide acetate, 11-dehydrosinulariolide, 11 episinulariolide, and sinulariolide. Their structures were elucidated by spectroscopic analysis, especially 2D-NMR and HR-ESI-MS. Compound 2 exhibited significant in vitro cytotoxic activity against human colon adenocarcinoma (WiDr) cell line. PMID- 20020463 TI - HS-SPME/GC analysis reveals the population variability of terpene contents in Juniperus communis needles. AB - Juniperus communis var. communis L. is an aromatic plant - typical boreal element of flora. In the extensive literature concerning J. communis, there is much data on the composition and the content of essential oil of needles and coneberries, but a detailed analysis of terpene distribution within and between populations is missing. A representative pool of 74 J. communis individuals originating from ten populations of Northern Poland was investigated in order to evaluate the intra- and interpopulational variability of the terpene pattern. Headspace solid-phase microextraction (HS-SPME) coupled with GC/MS and GC/FID was applied in achiral and enantioselective analysis. The majority of the samples (85%), despite different origin, were similar in the terpene pattern. High diversity of terpenes was observed within the populations and low diversity between them. High variation of enantiomeric composition was in accordance with large variation of individual compounds in general (achiral analysis). J. communis samples from Northern Poland could be distinguished by the alpha-pinene/sabinene ratio, and they were divided into three chemical races. PMID- 20020464 TI - Chemical composition of hydrodistilled essential oil of Artemisia incana (L.) Druce and antimicrobial activity against foodborne microorganisms. AB - The oil obtained by hydrodistillation from the aerial parts of Artemisia incana (L.) Druce from Turkey was analyzed by GC and GC/MS. Sixty-three compounds were characterized, representing 97.2% of the total components detected, and camphor (19.0%), borneol (18.9%), 1,8-cineole (14.5%), bornyl acetate (7.8%), camphene (4.9%), and alpha-thujone (4.8%) were identified as predominant components. The essential oil was also tested for its antimicrobial activity against 44 different foodborne microorganisms, including 26 bacteria, 15 fungi, and 3 yeast species. The essential oil of A. incana exhibited considerable inhibitory effects against all bacteria, fungi, and yeast species tested. However, the oil showed lower inhibitory activity against the tested bacteria than the reference antibiotics. PMID- 20020465 TI - In silico structural study of random amino acid sequence proteins not present in nature. AB - The three-dimensional structures of a set of 'never born proteins' (NBP, random amino acid sequence proteins with no significant homology with known proteins) were predicted using two methods: Rosetta and the one based on the 'fuzzy-oil drop' (FOD) model. More than 3000 different random amino acid sequences have been generated, filtered against the non redundant protein sequence data base, to remove sequences with significant homology with known proteins, and subjected to three-dimensional structure prediction. Comparison between Rosetta and FOD predictions allowed to select the ten top (highest structural similarity) and the ten bottom (the lowest structural similarity) structures from the ranking list organized according to the RMS-D value. The selected structures were taken for detailed analysis to define the scale of structural accordance and discrepancy between the two methods. The structural similarity measurements revealed discrepancies between structures generated on the basis of the two methods. Their potential biological function appeared to be quite different as well. The ten bottom structures appeared to be 'unfoldable' for the FOD model. Some aspects of the general characteristics of the NBPs are also discussed. The calculations were performed on the EUChinaGRID grid platform to test the performance of this infrastructure for massive protein structure predictions. PMID- 20020466 TI - Prenatal diagnosis and outcome of partial agenesis and hypoplasia of the corpus callosum. AB - OBJECTIVE: To present antenatal sonographic findings and outcome of fetuses with hypoplasia or partial agenesis of the corpus callosum. METHODS: The database of our ultrasound laboratory was searched retrospectively for cases of hypoplasia or partial agenesis of the corpus callosum suspected at antenatal neurosonography between 1998 and 2008 and confirmed by pathology or postnatal neuroimaging. In surviving infants, clinical follow-up had been arranged to assess neurodevelopmental outcome. RESULTS: Nineteen fetuses with callosal underdevelopment were identified at a median gestational age of 22 (range, 21-33) weeks and confirmed at follow-up, including 14 with partial agenesis and five with hypoplasia. Among the 14 fetuses with partial agenesis, there were additional brain findings in 10, including two with absent cavum septi pellucidi, four with mild isolated ventriculomegaly and four with cerebellar abnormalities, two of which also had ventriculomegaly. Pregnancy was terminated electively in seven of the cases with partial agenesis and there was one neonatal death. Among the six surviving infants, neurodevelopmental outcome was appropriate for age in three at follow up, including two cases with isolated partial agenesis of the corpus callosum. Among the five fetuses with prenatally diagnosed callosal hypoplasia, additional anomalies were present in four. Two cases were terminated electively and three were alive at the time of writing, with a median age of 3 years. Among them, apparently normal neurological development was observed in only one case. CONCLUSIONS: An antenatal diagnosis of callosal underdevelopment is possible by expert sonography. There is often association with other major anomalies. However, even in fetuses with apparently isolated findings, the prognosis is uncertain. PMID- 20020467 TI - Profiling fetal cardiac function in twin-twin transfusion syndrome. AB - OBJECTIVES: Cardiomyopathy in the recipient twin is a marker of severity in twin twin transfusion syndrome (TTTS), making it a potentially valuable tool for staging the disease. This study aimed to provide a quantitative description of cardiac function in the recipient twin. METHODS: Consecutive monochorionic pregnancies complicated with TTTS and treated by percutaneous laser coagulation underwent fetal echocardiography before surgery. An unsupervised classification analysis was conducted to identify groups of twins with similar cardiac profiles. The predictive value of the recipient twin's preoperative cardiac function based on these profiles was assessed, using perinatal death of at least one twin as the main outcome. The cardiac function profiles that we identified were compared with the current Quintero staging. RESULTS: A total of 107 pregnancies were included, with six of these lost to follow-up; 63/107 complete cases were available for multivariate description of the recipient's cardiac function. Three different preoperative cardiac profiles were identified with increasing right and left myocardial performance index, decreasing right and left shortening fraction, and increasing ductus venosus pulsatility index. Although the three groups represented progressive stages of the syndrome-related cardiomyopathy, no correlation was found with pregnancy outcome. Of Quintero Stage 1 cases, 55% showed significant alterations of cardiac function in the recipient twin. CONCLUSIONS: Progressive cardiomyopathy can be assessed quantitatively in the recipient twin and does not influence pregnancy outcome when fetoscopic laser coagulation is the first-line treatment. Compared with the current staging, cardiac profiling allows discrimination of cases with significant myocardial dysfunction. PMID- 20020469 TI - Bovine serum albumin-based magnetic nanocarrier for MRI diagnosis and hyperthermic therapy: a potential theranostic approach against cancer. PMID- 20020470 TI - A supramolecular network as sacrificial mask for the generation of a nanopatterned binary self-assembled monolayer. PMID- 20020468 TI - Upregulation of heme oxygenase-1 inhibits the maturation and mineralization of osteoblasts. AB - Heme-oxygenase-1 (HO-1), an important enzyme involved in vascular disease, transplantation, and inflammation, catalyzes the degradation of heme into carbon monoxide and biliverdin. It has been reported that overexpression of HO-1 inhibits osteoclastogenesis. However, the effect of HO-1 on osteoblast differentiation is still not clear. We here used adenoviral vector expressing recombinant human HO-1 and HO-1 inducer hemin to study the effects of HO-1 in primary cultured osteoblasts. The results showed that induction of HO-1 inhibited the maturation of osteoblasts including mineralized bone nodule formation, alkaline phosphatase activity and decreased mRNA expression of several differentiation markers such as alkaline phosphatase, osteocalcin, and RUNX2. Furthermore, downstream products of HO-1, bilirubin, carbon monoxide, and iron, are involved in the inhibitory action of HO-1. HO-1 can be induced by H(2)O(2), lipopolysaccharide and inflammatory cytokines such as TNF-alpha and IL-1beta in osteoblasts and also in STZ-induced diabetic mice. In addition, endogenous PPARgamma ligand, 15-deoxy-Delta(12,14)-prostaglandin-J2 (15d-PGJ2) markedly increased both mRNA and protein levels of HO-1 in osteoblasts via PI3K-Akt and MAPK pathways. Blockade of HO activity by ZnPP IX antagonized the inhibitory action on osteocalcin expression by hemin and 15d-PGJ2. Our results indicate that upregulation of HO-1 inhibits the maturation of osteoblasts and HO-1 may be involved in oxidative- or inflammation-induced bone loss. PMID- 20020471 TI - Describing electron correlation effects in the framework of the elongation method -elongation-MP2: formalism, implementation and efficiency. AB - The extension of the elongation method into description of electron correlation effects at ab intio level is presented. The formalism and implementation of the elongation-MP2 methodology is discussed. The results of calculations for model systems are presented to illustrate efficiency and accuracy of the method. Directions of the further development are highlighted. PMID- 20020472 TI - Coarse-grained model of nucleic acid bases. AB - Atomistic simulations of nucleic acids are prohibitively expensive and, consequently, reduced models of these compounds are of great interest in the field. In this work, we propose a physics-based coarse-grained model of nucleic acid bases in which each base is represented by several (3-5) interaction centers. van der Waals interactions are modeled by Lennard-Jones spheres with a 12-6 potential energy function. The charge distribution is modeled by a set of electric dipole moments located at the centers of the Lennard-Jones spheres. The method for computing the Lennard-Jones parameters, electric dipole moments (their magnitude and orientation) and positions of the interaction centers is described. Several models with different numbers of interaction centers were tested. The model with three-center cytosine, four-center guanine, four-center thymine, and five-center adenine satisfactorily reproduces the canonical Watson-Crick hydrogen bonding and stacking interaction energies of the all-atom AMBER model. The computation time with the coarse-grained model is reduced seven times compared with that of the all-atom model. PMID- 20020473 TI - An ab initio quantum mechanical charge field molecular dynamics simulation of a dilute aqueous HCl solution. AB - An ab initio quantum mechanical charge field (QMCF) molecular dynamics simulation has been performed to study the structural and dynamical properties of a dilute aqueous HCl solution. The solute molecule HCl and its surrounding water molecules were treated at Hartree-Fock level in conjunction with Dunning double-zeta plus polarization function basis sets. The simulation predicts an average H-Cl bond distance of 1.28 A, which is in good agreement with the experimental value. The H(HCl)...O(w) and Cl(HCl)...H(w) distances of 1.84 and 3.51 A were found for the first hydration shell. At the hydrogen site of HCl, a single water molecule is the most preferred coordination, whereas an average coordination number of 12 water molecules of the full first shell was observed for the chloride site. The hydrogen bonding at the hydrogen site of HCl is weakened by proton transfer reactions and an associated lability of ligand binding. Two proton transfer processes were observed in the QMCF MD simulation, demonstrating acid dissociation of HCl. A weak structure-making/breaking effect of HCl in water is recognized from the mean residence times of 2.1 and 0.8 ps for ligands in the neighborhood of Cl and H sites of HCl, respectively. PMID- 20020474 TI - Parameterization of the torsional potential for calix[4]arene-substituted poly(thiophene)s. AB - Three different strategies have been followed to develop the torsional force field parameters of the inter-ring dihedral angles for calix[4]arene-substituted poly(thiophene)s, a family of highly sensitive ion receptors. These procedures, which are based on the rotational profiles calculated using quantum mechanical methods, differ in the complexity of the model compounds and the processing applied to the quantum mechanical energies before the fitting. The performance of the three sets of developed parameters, which are essentially compatible with the General Amber Force Field, has been evaluated by computing the potential of mean forces for the inter-ring rotation of 2,2'-bithiophene, and its substituted analog bearing a calix[4]arene group in different environments. Finally, the ability of the new sets of torsional parameters to describe a calix[4]arene substituted poly(thiophene) in tetrahydrofuran solution has been checked using Molecular Dynamics simulations. Specifically, the molecular shape, the polymer conformation, and the effects of the Na(+) ions trapped in the cavity of the receptor have been examined. Although the potential derived from unsubstituted 2,2'-bithiophene is able to reproduce the experimental free energies of the minima, the overall results indicate that the parameters derived from the analog bearing a calix[4]arene group provide the best description of the systems under study. This should be attributed to the strong constraints found in complex substituted poly(thiophene)s, which require parameterization strategies able to capture all the interactions and phenomena involved in their inter-ring rotations. PMID- 20020476 TI - G protein-coupled receptors: the inside story. AB - Recent findings necessitate revision of the traditional view of G protein-coupled receptor (GPCR) signaling and expand the diversity of mechanisms by which receptor signaling influences cell behavior in general. GPCRs elicit signals at the plasma membrane and are then rapidly removed from the cell surface by endocytosis. Internalization of GPCRs has long been thought to serve as a mechanism to terminate the production of second messengers such as cAMP. However, recent studies show that internalized GPCRs can continue to either stimulate or inhibit cAMP production in a sustained manner. They do so by remaining associated with their cognate G protein subunit and adenylyl cyclase at endosomal compartments. Once internalized, the GPCRs produce cellular responses distinct from those elicited at the cell surface. PMID- 20020477 TI - Proteomics enhances evolutionary and functional analysis of reproductive proteins. AB - Reproductive proteins maintain species-specific barriers to fertilization, affect the outcome of sperm competition, mediate reproductive conflicts between the sexes, and potentially contribute to the formation of new species. However, the specific proteins and molecular mechanisms that underlie these processes are understood in only a handful of cases. Advances in genomic and proteomic technologies enable the identification of large suites of reproductive proteins, making it possible to dissect reproductive phenotypes at the molecular level. We first review these technological advances and describe how reproductive proteins are identified in diverse animal taxa. We then discuss the dynamic evolution of reproductive proteins and the potential selective forces that act on them. Finally, we describe molecular and genomic tools for functional analysis and detail how evolutionary data may be used to make predictions about interactions among reproductive proteins. PMID- 20020478 TI - Freedom of misinformation, but restriction of education...: public 'education' in science is frowned upon these days, but it is dangerous to write it off. PMID- 20020479 TI - Does CTCF mediate between nuclear organization and gene expression? AB - The multifunctional zinc-finger protein CCCTC-binding factor (CTCF) is a very strong candidate for the role of coordinating the expression level of coding sequences with their three-dimensional position in the nucleus, apparently responding to a "code" in the DNA itself. Dynamic interactions between chromatin fibers in the context of nuclear architecture have been implicated in various aspects of genome functions. However, the molecular basis of these interactions still remains elusive and is a subject of intense debate. Here we discuss the nature of CTCF-DNA interactions, the CTCF-binding specificity to its binding sites and the relationship between CTCF and chromatin, and we examine data linking CTCF with gene regulation in the three-dimensional nuclear space. We discuss why these features render CTCF a very strong candidate for the role and propose a unifying model, the "CTCF code," explaining the mechanistic basis of how the information encrypted in DNA may be interpreted by CTCF into diverse nuclear functions. PMID- 20020480 TI - Parallel evolution of segmentation by co-option of ancestral gene regulatory networks. AB - Different sources of data on the evolution of segmentation lead to very different conclusions. Molecular similarities in the developmental pathways generating a segmented body plan tend to suggest a segmented common ancestor for all bilaterally symmetrical animals. Data from paleontology and comparative morphology suggest that this is unlikely. A possible solution to this conundrum is that throughout evolution there was a parallel co-option of gene regulatory networks that had conserved ancestral roles in determining body axes and in elongating the anterior-posterior axis. Inherent properties in some of these networks made them easily recruitable for generating repeated patterns and for determining segmental boundaries. Phyla where this process happened are among the most successful in the animal kingdom, as the modular nature of the segmental body organization allowed them to diverge and radiate into a bewildering array of variations on a common theme. PMID- 20020481 TI - Accurate conformation-dependent molecular electrostatic potentials for high throughput in silico drug discovery. AB - The atom-centered partial charges-approximation is commonly used in current molecular modeling tools as a computationally inexpensive alternative to quantum mechanics for modeling electrostatics. Even today, the use of partial charges remains useful despite significant advances in improving the efficiency of ab initio methods. Here, we report on new parameters for the EEM and SFKEEM electronegativity equalization-based methods for rapidly determining partial charges that will accurately model the electrostatic potential of flexible molecules. The developed parameters cover most pharmaceutically relevant chemistries, and charges obtained using these parameters reproduce the B3LYP/cc pVTZ reference electrostatic potential of a set of FDA-approved drug molecules at best to an average accuracy of 13 +/- 4 kJ mol(-1); thus, equipped with these parameters electronegativity equalization-based methods rival the current best non-quantum mechanical methods, such as AM1-BCC, in accuracy, yet incur a lower computational cost. Software implementations of EEM and SFKEEM, including the developed parameters, are included in the conformer-generation tool BALLOON, available free of charge at http://web.abo.fi/fak/mnf/bkf/research/johnson/software.php. PMID- 20020482 TI - A molecular mechanics model for imatinib and imatinib:kinase binding. AB - Imatinib is an important anticancer drug, which binds specifically to the Abl kinase and blocks its signalling activity. To model imatinib:protein interactions, we have developed a molecular mechanics force field for imatinib and four close analogues, which is consistent with the CHARMM force field for proteins and nucleic acids. Atomic charges and Lennard-Jones parameters were derived from a supermolecule ab initio approach. We considered the ab initio energies and geometries of a probe water molecule interacting with imatinib fragments at 32 different positions. We considered both a neutral and a protonated imatinib. The final RMS deviation between the ab initio and force field energies, averaged over both forms, was 0.2 kcal/mol. The model also reproduces the ab initio geometry and flexibility of imatinib. To apply the force field to imatinib:Abl simulations, it is also necessary to determine the most likely imatinib protonation state when it binds to Abl. This was done using molecular dynamics free energy simulations, where imatinib is reversibly protonated during a series of MD simulations, both in solution and in complex with Abl. The simulations indicate that imatinib binds to Abl in its protonated, positively-charged form. To help test the force field and the protonation prediction, we did MD free energy simulations that compare the Abl binding affinities of two imatinib analogs, obtaining good agreement with experiment. Finally, two new imatinib variants were considered, one of which is predicted to have improved Abl binding. This variant could be of interest as a potential drug. PMID- 20020483 TI - A first principle study of the structural, vibrational and electronic properties of tetrathiafulvalene adsorbed on Ag(110) and Au(110) surfaces. AB - We have studied the adsorption properties of a charge donor organic molecule, tetrathiafulvalene (TTF), on the (110) surfaces of silver and gold by means of the generalized gradient approach of the density functional theory using periodic slab models. This molecule is the core building block of a host of molecular materials exhibiting extremely reach phase diagrams with a variety of ground states. The interfaces formed with metallic surfaces have received only limited attention, despite of their relevance. We have determined the stable adsorption sites for two unit cells representing high and low coverage, which are determinant for the adsorption properties of TTF on the surface. The preferential chemisorption is via the direct interaction of sulfur atoms with the Ag or Au atoms on top sites. All adsorbed TTF are more stable than gas phase TTF. The simulation of the vibrational spectra has permitted us to find the fingerprints of these structures to characterize them on this surface. The donor nature of TTF induces charge transfer to the metallic surfaces. PMID- 20020484 TI - Electron localizability indicators ELI-D and ELIA for highly correlated wavefunctions of homonuclear dimers. I. Li2, Be2, B2, and C2. AB - Electron localizability indicators based on the parallel-spin electron pair density (ELI-D) and the antiparallel-spin electron pair density (ELIA) are studied for the correlated ground-state wavefunctions of Li(2), Be(2), B(2), and C(2) diatomic molecules. Different basis sets and reference spaces are used for the multireference configuration interaction method following the complete active space calculations to investigate the local effect of electron correlation on the extent of electron localizability in position space determined by the two functionals. The results are complemented by calculations of effective bond order, vibrational frequency, and Laplacian of the electron density at the bond midpoint. It turns out that for Li(2), B(2), and C(2) the reliable topology of ELI-D is obtained only at the correlated level of theory. PMID- 20020485 TI - Thrombospondin type I domain containing 7A (THSD7A) mediates endothelial cell migration and tube formation. AB - Angiogenesis is a highly organized process controlled by a series of molecular events. While much effort has been devoted to identifying angiogenic factors and their reciprocal receptors, far less information is available on the molecular mechanisms underlying directed endothelial cell migration. To search for novel proteins that participate in this process, we used the serial analysis of gene expression (SAGE) transcript profiling approach to identify genes that are selectively expressed in endothelial cells (ECs). Two EC SAGE libraries were constructed from human umbilical vein and artery ECs to enable data-mining against other non-ECs. A novel endothelial protein, Thrombospondin Type I Domain Containing 7A (THSD7A), with preferential expression in placenta vasculature and in human umbilical vein endothelial cells (HUVECs) was identified and targeted for further characterization. Overexpression of a THSD7A carboxyl-terminal fragment in HUVECs inhibited cell migration and disrupted tube formation, while suppression of THSD7A expression enhanced HUVEC migration and tube formation. Immunohistological analysis revealed that THSD7A was expressed at the leading edge of migrating HUVECs, and it co-localized with alpha(V)beta(3) integrin and paxillin. This distribution was dispersed from focal adhesions after disruption of the actin cytoskeleton, suggesting the involvement of THSD7A in cytoskeletal organization. Our results show that THSD7A is a novel placenta endothelial protein that mediates EC migration and tube formation, and they highlight its potential as a new target for anti-angiogenic therapy. PMID- 20020486 TI - DFT study of the asymmetric nitroaldol (Henry) reaction catalyzed by a dinuclear Zn complex. AB - We report the mechanism of asymmetric nitroaldol (Henry) reaction catalyzed by a dinuclear Zn complex using density functional theory. The experimentally proposed catalytic cycle is validated, in which the first step is the deprotonation of nitromethane by the ethyl anion of the catalyst, subsequently a C-C bond formation step, and then the protonation of the resulting alkoxide. Three mechanistic scenarios (differing in binding modes) have been considered for the C C bond formation step. The origin of the enantioselectivity is discussed. Our calculations supported that the S configurations are the major products, which is in agreement with the experimental observations. PMID- 20020488 TI - Mosaic trisomy 22: five new cases with variable outcomes. Implications for genetic counselling and clinical management. PMID- 20020487 TI - 20,23-dihydroxyvitamin D3, novel P450scc product, stimulates differentiation and inhibits proliferation and NF-kappaB activity in human keratinocytes. AB - We have examined effects of the 20,23-dihydroxyvitamin D3 (20,23(OH)2D3), on differentiation and proliferation of human keratinocytes and the anti inflammatory potential of 20,23(OH)2D3 from its action on nuclear factor-kappaB (NF-kappaB). 20,23(OH)2D3 inhibited growth of keratinocytes with a potency comparable to that for 1,25-dihydroxyvitamin D3 (1,25(OH)2D3). Cell cycle analysis showed that this inhibition was associated with G1/G0 and G2/M arrests. 20,23(OH)2D3 stimulated production of involucrin mRNA and inhibited production of cytokeratin 14 mRNA in a manner similar to that seen for 1,25(OH)2D3. Flow cytometry showed that these effects were accompanied by increased involucrin protein expression, and an increase in the cell size and granularity. Silencing of the vitamin D receptor (VDR) by corresponding siRNA abolished the stimulatory effect on involucrin gene expression demonstrating an involvement of VDR in 20,23(OH)2D3 action. This mode of action was further substantiated by stimulation of CYP24 gene expression and stimulation of the CYP24 promoter-driven reporter gene activity. 20,23(OH)2D3 displayed several fold lower potency for induction of CYP24 gene expression than 1,25(OH)2D3. Finally, 20,23(OH)2D3 inhibited the transcriptional activity of NF-kappaB in keratinocytes as demonstrated by EMSA, NF-kappaB-driven reporter gene activity assays and measurements of translocation of p65 from the cytoplasm to the nucleus. These inhibitory effects were connected with stimulation of the expression of IkappaBalpha with subsequent sequestration of NF-kappaB in the cytoplasm and consequent attenuation of transcriptional activity. In summary, we have characterized 20,23(OH)2D3 as a novel secosteroidal regulator of keratinocytes proliferation and differentiation and a modifier of their immune activity. PMID- 20020491 TI - Gastrin suppresses the interdependent expression of p16 and anion exchanger 1 favoring growth inhibition of gastric cancer cells. AB - Our previous studies demonstrated that expression and interaction of p16 with anion exchanger 1 (AE1) in gastric cancer cells is correlated with progression and shorter survival of the cancer. In this article, the effects of gastrin on p16 and AE1 and its implication in prevention and treatment of gastric cancer were studied by molecular biology techniques, animal experiment and clinical analysis. The results showed that expression of p16 in human gastric body carcinoma was downregulated along with the progression of the cancer, suggesting the reverse correlations between gastrin and p16 in vivo. Further experiments indicated that gastrin suppressed the expression of p16 via the p16 promoter and thereafter resulted in the degradation of AE1 in gastric cancer cells. Silencing of AE1 or p16 significantly inhibited the proliferation of the cancer cells. Using a xenograft tumor model in nude mice, we showed that experimental systemic hypergastrinemia induced by the administration of omeprazole led to decreased expression of AE1 and p16 as well as to a marked growth inhibition of SGC7901 tumors. It is concluded that a moderate plasma gastrin level is beneficial to the growth inhibition of gastric cancer by suppressing the expression of AE1 and p16. This finding may have an important implication for the prevention and treatment of cancers arise in the gastric antrum. PMID- 20020490 TI - TGF-beta inactivation and TGF-alpha overexpression cooperate in an in vivo mouse model to induce hepatocellular carcinoma that recapitulates molecular features of human liver cancer. AB - Hepatocellular carcinoma (HCC) results from the cumulative effects of deregulated tumor suppressor genes and oncogenes. The tumor suppressor and oncogenes commonly affected include growth factors, receptors and their downstream signaling pathway components. The overexpression of transforming growth factor alpha (TGF-alpha) and the inhibition of TGF-beta signaling are especially common in human liver cancer. Thus, we assessed whether TGF-alpha overexpression and TGF-beta signaling inactivation cooperate in hepatocarcinogenesis using an in vivo mouse model, MT1/TGFa;AlbCre/Tgfbr2(flx/flx) mice ("TGFa;Tgfbr2(hepko)"), which overexpresses TGF-alpha and lacks a TGF-beta receptor in the liver. TGF-beta signaling inactivation did not alter the frequency or number of cancers in mice with overexpression of TGF-alpha. However, the tumors in the TGFa;Tgfbr2(hepko) mice displayed increased proliferation and increased cdk2, cyclin E and cyclin A expression as well as decreased Cdkn1a/p21 expression compared to normal liver and compared to the cancers arising in the TGF-alpha overexpressing mice with intact TGF-beta receptors. Increased phosphorylated ERK1/2 expression was also present in the tumors from the TGFa;Tgfbr2(hepko) mice and correlated with downregulated Raf kinase inhibitor protein expression, which is a common molecular event in human HCC. Thus, TGF-beta signaling inactivation appears to cooperate with TGF-alpha in vivo to promote the formation of liver cancer that recapitulates molecular features of human HCC. PMID- 20020492 TI - Epidermal growth factor receptor expression affects the efficacy of the combined application of saponin and a targeted toxin on human cervical carcinoma cells. AB - Cervical cancer is the second most common cancer in women worldwide. Targeting the epidermal growth factor receptor (EGFR) is a very promising approach since it is overexpressed in about 90% of cervical tumors. Here, we quantified the toxic effect of SE, a targeted toxin consisting of epidermal growth factor (EGF) as targeting moiety and the plant toxin saporin-3, on 3 common human cervical carcinoma cell lines (HeLa, CaSki and SiHa) and recently established lines (PHCC1 and PHCC2) from 2 different individuals. A human melanocytic and a mouse cell line served as negative control. Additionally, we combined SE with saponinum album, a saponin composite from Gypsophila paniculata, which exhibited synergistic properties in previous studies. The cell lines, except for SiHa cells, revealed high sensitivity to SE with 50% cell survival in the range of 5 24.5 nM. The combination with saponin resulted in a remarkable enhancement of cytotoxicity with enhancement factors ranging from 9,000-fold to 2,500,000-fold. The cytotoxicity of SE was clearly target receptor specific since free EGF blocks the effect and saporin-3 alone was considerably less toxic. For all cervical carcinoma cell lines, we evinced a clear correlation between EGFR expression and SE sensitivity. Our data indicate a potential use of targeted toxins for the treatment of cervical cancer. In particular, the combination with saponins is a promising approach since efficacy is drastically improved. PMID- 20020493 TI - Pattern of declining hemoglobin concentration before cancer diagnosis. AB - Although anemia is widely considered an early sign of malignant disease, little is known about the pattern of hemoglobin decline before diagnosis. As an approach to understanding the duration of the preclinical phase of different types of malignant diseases, we investigated prediagnostic hemoglobin concentration changes in a large cohort of blood donors. Using a nested case-control design, we analyzed a population-based cohort comprising 1.1 million Scandinavian blood donors with complete follow-up through record linkage to population and cancer registers. A total of 16,375 cancer cases were identified, for whom we selected 161,995 controls. We used conditional logistic regression to estimate the risk of cancer in relation to hemoglobin concentration during the 5 years preceding the cancer diagnosis. Hemoglobin concentration decline began already 3 years before diagnosis of stomach cancer, multiple myeloma, and lymphatic leukemia; 2 years before diagnosis of small intestinal and colon cancer as well as of Hodgkin lymphoma. A decline was evident during the last year for non-Hodgkin lymphoma and myeloid/monocytic leukemia, whereas no change was found for cancer of the esophagus, breast or prostate. In conclusion, in this study, we have demonstrated that the pattern of declining hemoglobin concentration before cancer diagnosis varies considerably between malignancies without being a suitable screening tool for any of them. For some malignancies, however, the long duration of hemoglobin decline before clinical diagnosis suggests a substantial lead-time with systemic effects, during which earlier diagnosis should be achievable by emerging diagnostic tools. PMID- 20020494 TI - Inhibition of immune responses by Dasatinib may account for its different effects on neuroblastoma between in vitro and in vivo. PMID- 20020495 TI - RACK1: A superior independent predictor for poor clinical outcome in breast cancer. AB - We aimed to investigate the expression of RACK1 in breast cancer, evaluate its role in predicting prognosis and compare with commonly used biomarkers: Ki67, ER, PR and HER-2 for patients with breast cancer. The RACK1 expression and its clinical significance were examined in 160 breast carcinoma patients using immunohistochemistry. Correlations of RACK1 expression with other commonly used biomarkers and survival analyses were assessed. Immunohistochemistry results showed that the number of RACK1 cases scoring 0, 1, and 2 were 66, 54, and 40, respectively. RACK1 staining was strongly related to clinical stage, histological grade, Ki67, ER, PR and HER-2 (all p < 0.05). Consistently, all of the cases exhibiting RACK1 staining score 0 were survivors, whereas the majority (55.0%) of those exhibiting RACK1 staining score 2 were deaths. Kaplan-Meier survival analysis of 160 cases revealed a correlation between higher RACK1 expression levels and shorter overall survival times (p < 0.001). Univariate and multivariate analyses revealed that RACK1, tumor size, lymph node metastasis, and HER-2 were independent prognostic factors (all p < 0.05). Interestingly, receiver operator characteristic (ROC) curves showed that the ROC areas for RACK1, Ki67, ER, PR and HER-2 were 0.833, 0.766, 0.446, 0.387, and 0.689, respectively, and the superiority of RACK1 in sensitivity and specificity as biomarker was demonstrated. To our knowledge, it is the first time to investigate the expression of RACK1, and identified that RACK1 is a superior independent biomarker for diagnosis and prognosis comparing with currently widely used diagnostic index in breast carcinoma. PMID- 20020496 TI - HIF-1alpha is an unfavorable determinant of relapse in gastric cancer patients who underwent curative surgery followed by adjuvant 5-FU chemotherapy. AB - Among several chemotherapeutic agents, 5-fluorouracil (5-FU) has been widely used as a key drug in adjuvant chemotherapy for gastric cancer. However, no reliable marker, which predicts the response to 5-FU in an adjuvant setting, has been identified. Hypoxia-induced drug resistance, via upregulation of HIF-1alpha, is a major obstacle in the development of effective cancer therapy. However, few clinical studies have so far assessed the relationship between the HIF-1alpha expression and the chemo-resistance of gastric cancer patients in an adjuvant setting. We established 2 HIF-1alpha knockdown gastric cancer cell lines in order to clarify the role of HIF-1alpha in chemo-resistance against 5-FU. Furthermore, expression of HIF-1alpha was immunohistochemically assessed in 91 resected specimens. Sixty-four of 91 patients received 5-FU adjuvant chemotherapy after surgery. HIF-1alpha expression was associated with the significantly shorter relapse-free survival and disease-specific survival in the 64 patients of adjuvant group (p = 0.026, 0.014, respectively), but not in the 27 of surgery group. Multivariate analysis showed that HIF-1alpha was an independent risk factor for relapse in 64 patients in the adjuvant group (p = 0.029). In conclusion, the current study confirmed, for the first time that HIF-1alpha expression is an independent risk factor for relapse in high-risk gastric cancer patients who underwent curative surgery followed by adjuvant 5-FU chemotherapy. A favorable effect of 5-FU might therefore be expected in patients that do not express HIF-1alpha, whereas, other types of chemotherapy or additional treatments, such as HIF-1alpha inhibitors, should be considered in patients that do express HIF-1alpha. PMID- 20020497 TI - miR-212 is downregulated and suppresses methyl-CpG-binding protein MeCP2 in human gastric cancer. AB - To clarify the role of micro (mi) RNAs in gastric carcinogenesis, we studied the expression and function of miRNAs in gastric carcinoma (GC) cells. Initially, we performed microarray analysis using total RNA from 3 human GC cell lines and noncancerous gastric tissue. Among the downregulated miRNAs in GC cells, miR-212 expression was decreased in all 8 GC cell lines examined and a significant decrease of miR-212 expression in human primary GC tissues was also observed in 6 of 11 cases. Transfection of the precursor miR-212 molecule induced decreased growth of 3 GC cell lines. Using 3 different databases, methyl-CpG-binding protein MeCP2 was postulated to be a target of miR-212. As seen on reporter assaying, miR-212 repressed the construct with the MECP2 3'-UTR. Ectopic expression of miR-212 repressed expression of the MeCP2 protein but not the MECP2 mRNA level. These data suggest that downregulation of miR-212 may be related to gastric carcinogenesis through its target genes, such as MECP2. PMID- 20020498 TI - The protease inhibitor, elafin, induces p53-dependent apoptosis in human melanoma cells. AB - Expression of the protease inhibitor elafin is deregulated in several human cancers. However, functions of the protein in cancer are yet to be established. Here, we show that elafin elicits pro-apoptotic effects in melanoma cells but not in normal melanocytes. Elafin triggered the intrinsic apoptotic pathway as evidenced by the increased caspase 9 activity and unaltered caspase 8 activity. Caspase 9-specific siRNA, but not caspase 8-specific siRNA, dramatically abrogated elafin-induced apoptosis. Elevated level of p53 was observed, resulting in increased transcriptional activation and consequent expression of downstream effector molecules (Bax, Puma, Noxa, p21). Moreover, the apoptotic effect of elafin was inhibited by p53-specific siRNA and the p53 inhibitor pifithrin-alpha. Elafin treatment of xenograft mice of melanoma cells led to significantly smaller tumor sizes compared with those of untreated control mice. Immunohistochemical analysis revealed decreased elafin expression in melanoma tissue specimens. Western blot and reverse transcription analyses indicated transcriptional repression of the elafin gene in melanoma cells. Our results collectively indicate that elafin induces apoptosis in melanoma cells through a p53-dependent intrinsic apoptotic pathway, and that repression of elafin expression in melanoma may contribute to disease progression. PMID- 20020500 TI - Toll-like receptor 2-mediated peptidoglycan uptake by immature intestinal epithelial cells from apical side and exosome-associated transcellular transcytosis. AB - Peptidoglycan (PGN) is a potent immune adjuvant derived from bacterial cell walls. Previous investigations suggest that intestinal epithelium may absorb PGN from the lumen. Nonetheless, how PGN is taken up and crosses intestinal epithelium remains largely unclear. Here, we first characterized PGN transport in vitro using IEC-18 and HT29-CL19A cells, which represent less mature epithelial cells in intestinal crypts. With fluorescent microscopy, we visualized internalization of dual-labeled PGN by enterocytes. Engulfed PGN was found to form a complex with PGN recognition protein-3, which may facilitate delivering PGN in vivo. Utilizing electronic microscopy, we revealed that uptake of apical PGN across intestinal epithelial monolayers was involved in phagocytosis, multivesicular body formation, and exosome secretion. We also studied transport of PGN using the transwell system. Our data indicated that apically loaded PGN was exocytosed to the basolateral compartment with exosomes by HT29-CL19A cells. The PGN-contained basolateral exosome extracts induced macrophage activation. Through gavaging mice with labeled PGN, we found that luminal PGN was taken up by columnar epithelial cells in crypts of the small intestine. Furthermore, we showed that pre-confluent immature but not post-confluent mature C2BBe1 cells engulfed PGN via a toll-like receptor 2-dependent manner. Together, our findings suggest that (1) crypt-based immature intestinal epithelial cells play an important role in transport of luminal PGN over the intestinal epithelium; and (2) luminal PGN is transcytosed across intestinal epithelia via a toll-like receptor 2-mediated phagocytosis-multivesicular body-exosome pathway. The absorbed PGN and its derivatives may facilitate maintenance of intestinal immune homeostasis. PMID- 20020499 TI - Toward a population genetic framework of developmental evolution: the costs, limits, and consequences of phenotypic plasticity. AB - Adaptive phenotypic plasticity allows organisms to cope with environmental variability, and yet, despite its adaptive significance, phenotypic plasticity is neither ubiquitous nor infinite. In this review, we merge developmental and population genetic perspectives to explore costs and limits on the evolution of plasticity. Specifically, we focus on the role of modularity in developmental genetic networks as a mechanism underlying phenotypic plasticity, and apply to it lessons learned from population genetic theory on the interplay between relaxed selection and mutation accumulation. We argue that the environmental specificity of gene expression and the associated reduction in pleiotropic constraints drive a fundamental tradeoff between the range of plasticity that can be accommodated and mutation accumulation in alternative developmental networks. This tradeoff has broad implications for understanding the origin and maintenance of plasticity and may contribute to a better understanding of the role of plasticity in the origin, diversification, and loss of phenotypic diversity. PMID- 20020501 TI - DNA supercoiling helps to unlink sister duplexes after replication. AB - DNA supercoiling is one of the mechanisms that can help unlinking of newly replicated DNA molecules. Although DNA topoisomerases, which catalyze the strand passing of DNA segments through one another, make the unlinking problem solvable in principle, it remains difficult to complete the process that enables the separation of the sister duplexes. A few different mechanisms were developed by nature to solve the problem. Some of the mechanisms are very intuitive while the others, like topology simplification by type II DNA topoisomerases and DNA supercoiling, are not so evident. A computer simulation and analysis of linked sister plasmids formed in Escherichia coli cells with suppressed topoisomerase IV suggests an insight into the latter mechanism. PMID- 20020503 TI - Design and testing of biological scaffolds for delivering reparative cells to target sites in the lung. AB - This study summarizes the development and testing of a scaffold to promote engraftment of cells in the distal lung. A fibrinogen-fibronectin-vitronectin hydrogel (FFVH) was developed and optimized with respect to its mechanical and biological properties for this application. In vitro, FFVH scaffolds promoted attachment, histiotypic growth and expression of basement membrane proteins by primary ovine lung mesenchymal cells derived from lung biopsies. In vivo testing was then performed to assess the ability of FFVHs to promote cell engraftment in the sheep lung. Treatment with autologous cells delivered using FFVH was clinically well tolerated. Cells labelled with a fluorescent dye (PKH-26) were detected at treatment sites after 1 month. Tissue mass (assessed by CT imaging) and lung perfusion (assessed by nuclear scintigraphy) were increased at emphysema test sites. Post-treatment histology demonstrated cell proliferation and increased elastin expression without scarring or collapse. No treatment-related pathology was observed at healthy control sites. FFVH scaffolds promote cell attachment, spreading and extracellular matrix expression in vitro and apparent engraftment in vivo, with evidence of trophic effects on the surrounding tissue. Scaffolds of this type may contribute to the development of cell-based therapies for patients with end-stage pulmonary diseases. PMID- 20020504 TI - Ex vivo expansion of human mesenchymal stem cells: a more effective cell proliferation kinetics and metabolism under hypoxia. AB - The low bone marrow (BM) MSC titers demand a fast ex vivo expansion process to meet the clinically relevant cell dosage. Attending to the low oxygen tension of BM in vivo, we studied the influence of hypoxia on human BM MSC proliferation kinetics and metabolism. Human BM MSC cultured under 2% (hypoxia) and 20% O(2) (normoxia) were characterized in terms of proliferation, cell division kinetics and metabolic patterns. BM MSC cultures under hypoxia displayed an early start of the exponential growth phase, and cell numbers obtained at each time point throughout culture were consistently higher under low O(2), resulting in a higher fold increase after 12 days under hypoxia (40 +/- 10 vs. 30 +/- 6). Cell labeling with PKH26 allowed us to determine that after 2 days of culture, a significant higher cell number was already actively dividing under 2% compared to 20% O(2) and BM MSC expanded under low oxygen tension displayed consistently higher percentages of cells in the latest generations (generations 4-6) until the 5th day of culture. Cells under low O(2) presented higher specific consumption of nutrients, especially early in culture, but with lower specific production of inhibitory metabolites. Moreover, 2% O(2) favored CFU-F expansion, while maintaining BM MSC characteristic immunophenotype and differentiative potential. Our results demonstrated a more efficient BM MSC expansion at 2% O(2), compared to normoxic conditions, associated to an earlier start of cellular division and supported by an increase in cellular metabolism efficiency towards the maximization of cell yield for application in clinical settings. PMID- 20020505 TI - Fibroblasts/myofibroblasts that participate in cutaneous wound healing are not derived from circulating progenitor cells. AB - Dermal fibroblasts/myofibroblasts involved in the wound healing are thought to originate from the resident fibroblast progenitors. To test the hypothesis of an extra dermal origin of the dermal fibroblasts/myofibroblasts, bone marrow (BM) transplantation and parabiosis experiments were initiated utilizing a collagen promoter green fluorescent protein (GFP) reporter transgene as a visible marker for dermal fibroblasts/myofibroblasts. BM transplantation experiments using BM from Col3.6GFPsapph transgenic mice showed no evidence that BM derived progenitors differentiated into dermal fibroblasts/myofibroblasts at the wound site. Rather the GFP positive cells (GFP+) observed at the wound site were not dermal fibroblasts/myofibroblasts but immune cells. These GFP+ cells were also detected in the lung and spleen. Furthermore, GFP+ fibroblasts were not detected in primary dermal fibroblast cultures initiated from BM chimeras. Using the same transgenic mice, parabiotic pairs were generated. One partner in the parabiosis carried a GFP expressing transgene while the other partner was a non-transgenic C57BL/6 mouse. Similar to the BM transplantation experiments, GFP+ immune cells were detected in the wound of the non-transgenic parabiont, however, GFP expressing dermal fibroblasts/myofibroblasts were not observed. Collectively, these data suggest that dermal fibroblast/myofibroblast progenitors do not readily circulate. The expression of the Col3.6GFPsapph in the hematopoietic cells confirmed that our methods were sensitive enough to detect Col3.6GFP expressing dermal fibroblasts derived from the peripheral circulation if they had originated in the BM. PMID- 20020506 TI - Cytometry and DNA ploidy: clinical uses and molecular perspective in gastric and lung cancer. AB - Flow cytometry is one of the most powerful and specific methods used for the integrated study of the molecular and morphological events occurring during cell proliferation. Many methods have been described for investigating this process. Several cell cycle regulators controlling the correct entry and progression through the cell cycle are altered in tumors. In fact, in most, if not all, human cancers there is a deregulated control of G1 phase progression, the period when cells decide if they will start proliferation or stay quiescent. Cytometry (flow and image) is able to analyze DNA content thanks to the use of the same "molecule" conjugates with a fluorochrome that permits to identify DNA content of single cell in a sample. Most important results of studies on DNA ploidy have been reviewed during the last years and as a result the analyses of DNA ploidy in cancer may provide clinically useful information on diagnostic, therapeutic and prognostic aspects. In fact, aneuploid cancer has a high proliferative activity and a metastatic or invasive potential, markers of a poor prognosis. Multiparametric flow cytometry should allow the simultaneous determination of morphology, phenotype, intracellular protein expression, and status of chromatin and DNA. Evaluating if a particular protein is responsible for the aggressiveness of cancer, or the alteration of DNA content, or if the activation of its state is the cause of rapid growth of cancer cells, is very important and it can facilitate the clinical treatment of patients. PMID- 20020508 TI - Human cleft lip and palate fibroblasts and normal nicotine-treated fibroblasts show altered in vitro expressions of genes related to molecular signaling pathways and extracellular matrix metabolism. AB - Nonsyndromic cleft lip with or without cleft palate (CLP) is a frequent craniofacial malformation caused by both genetic and environmental factors. Maternal smoking during pregnancy is a known risk factor, due to the teratogenic role of nicotine. To assess and compare the impact of CLP and nicotine, we studied the quantitative expression of genes involved in signaling pathways and extracellular matrix (ECM) metabolism in human normal nicotine-treated (NicN) and CLP fibroblasts compared to normal control (CTRL) cells. Palatal fibroblast cultures from seven CLP children and seven age-matched CTRL subjects were established and subconfluent cells incubated for 24 h without (CTRL and CLP fibroblasts) or with (NicN fibroblasts) 0.6 mM nicotine. Gene expressions were analyzed by real-time quantitative PCR. For the first time, a regulated cholinergic signaling in our human fibroblasts in vitro was demonstrated. Members of TGF-beta, retinoic acid (RA), and GABA-ergic signaling systems were also differently regulated. Among the ECM genes, fibronectin, syndecan, integrin alpha2, and MMP13 genes were concordantly modulated, while integrin beta5, and decorin genes were discordantly modulated. Interestingly, nicotine treatment regulated gene expressions of CD44 and CLPTM1, two candidate genes for CLP. Our findings show a positive association between nicotine treatment and CLP phenotype. Results suggest that nicotine deranges normal palate development, which might contribute to the development of a CLP malformative phenotype, through the impairment of some important signaling systems and ECM composition. PMID- 20020507 TI - Genomic organization of microRNAs. AB - microRNAs (miRNAs) are small ( approximately 22 nt) noncoding RNAs that have been shown to regulate gene expression post-transcriptionally. They function by pairing with the 3' UTR of target mRNAs and repressing translation or by targeting the mRNA for degradation. miRNAs are involved in diverse aspects of development, maintenance, and disease, and are largely evolutionarily conserved in metazoans. Searching the genomes of organisms from viruses to worms to humans has revealed potentially thousands of miRNA genes. Understanding the patterns of genomic organization between species cannot only help to refine tools to identify new miRNAs, but also provide insight into miRNA biogenesis and function. PMID- 20020509 TI - Vertebral body innervation: Implications for pain. AB - Vertebral fractures often cause intractable pain. To define the involvement of vertebral body innervation in pain, we collected specimens from male and female patients during percutaneous kyphoplasty, a procedure used for reconstruction of the vertebral body. Specimens were taken from 31 patients (9 men and 22 women) suffering high-intensity pain before surgery. In total, 1,876 histological preparations were obtained and analysed. Immunohistochemical techniques were used to locate the nerves in the specimens. The nerve fibres were labelled by indirect immunofluorescence with the primary antibody directed against Protein Gene Product 9.5 (PGP 9.5), a pan-neuronal marker; another primary antibody directed against type IV collagen (Col IV) was used to identify vessels and to determine their relationship with vertebral nerve fibres. The mean percentage of samples in which it was possible to identify nerve fibres was 35% in men and 29% in women. The percentages varied depending on the spinal level considered and the sex of the subject, nerve fibres being mostly present around vessels (95%). In conclusion, there is scarce innervation of the vertebral bodies, with a clear prevalence of fibres located around vessels. It seems unlikely that this pattern of vertebral body innervation is involved in vertebral pain or in pain relief following kyphoplasty. PMID- 20020510 TI - Differential expression of interleukin-17 family cytokines in intact and complicated human atherosclerotic plaques. AB - In addition to the classical TH1 and TH2 cytokines, members of the recently identified IL-17 cytokine family play an important role in regulating cellular and humoral immune responses. At present nothing is known about the role of these cytokines in atherosclerosis. Expression of IL-17A, -E and -F was investigated in atherosclerotic tissue by rtPCR and immunohistochemistry. IL-17E and its receptor were further studied in cultured smooth muscle cells and endothelial cells, using rtPCR and western blot. rtPCR showed that IL-17A, -E and -F were expressed in the majority of plaques under investigation. IL-17A/F was expressed by mast cells in all stages of plaque development. IL-17A/F(+) neutrophils were always observed in complicated plaques, but hardly in intact lesions. IL-17A/F(+) Tcells ('TH17') were never observed. IL-17E was expressed by smooth muscle cells and endothelial cells in both normal and atherosclerotic arteries, and in advanced plaques also extensively by mature B cells. Cultured smooth muscle cells and endothelial cells were found to express both IL-17E and its functional receptor (IL-17RB). The constitutive expression of IL-17E by resident plaque cells, and the additional presence of IL-17E(+) B cells and IL-17A/F(+) neutrophils in advanced and complicated plaques indicates a complex contribution of IL-17 family cytokines in human atherosclerosis, depending on the stage and activity of the disease. PMID- 20020511 TI - Selective blockade of endothelial NF-kappaB pathway differentially affects systemic inflammation and multiple organ dysfunction and injury in septic mice. AB - Endothelium has long been considered both a source and a target of systemic inflammation. However, to what extent endothelial activation contributes to systemic inflammation remains unclear. This study addresses the relative contribution of endothelial activation to systemic inflammation and multiple organ dysfunction and injury (MOD/I) in an E. coli peritonitis model of sepsis. We prevented endothelial activation using transgenic (TG) mice that conditionally overexpress a mutant I-kappaBalpha, a NF-kappaB inhibitor, selectively on endothelium. TG mice and their transgene negative littermates (WT) were injected with saline or E. coli (10(8) CFU per mouse). At 7 h after E. coli infection, markers of systemic inflammation, endothelial activation, and MOD/I were assessed. WT-E. coli mice showed significantly increased serum levels of TNF alpha, IL-1beta, IFN-gamma, IL-6, KC, and MCP-1; tissue levels of TNF-alpha, IL 6, KC, MCP-1, ICAM-1, and VCAM-1; endothelial leakage index in heart, lungs, liver, and kidney; significantly increased serum levels of AST, ALT, BUN, and creatinine; and increased mortality. Blockade of NF-kappaB-mediated endothelial activation in TG mice had no effects on serum levels of TNF-alpha, IL-1beta, IFN gamma, IL-6, KC, and MCP-1 (markers of systemic inflammation), and tissue levels of TNF-alpha, IL-6, KC, and MCP-1, but significantly reduced tissue levels of ICAM-1 and VCAM-1 (markers of endothelial inflammation and activation) in those four organs. TG-E. coli mice displayed reversed endothelial leakage index; reduced serum levels of AST, ALT, BUN, and creatinine; and improved survival. Our data demonstrate that endothelial NF-kappaB-driven inflammatory response contributes minimally to systemic inflammation, but plays a pivotal role in septic MOD/I, suggesting that endothelium is mainly a target rather than a source of systemic inflammation. PMID- 20020512 TI - Age-related susceptibility to Streptococcus pyogenes infection in mice: underlying immune dysfunction and strategy to enhance immunity. AB - Epidemiological studies have shown that the elderly are at higher risk of severe Streptococcus pyogenes infections. In this study, we used a mouse model that displays the age-related loss of resistance to S. pyogenes infection seen in humans to investigate the impaired immune mechanism underlying the age-associated susceptibility to this pathogen. Young (2-3 months old) and aged (>20 months old) BALB/c mice were subcutaneously or intravenously inoculated with S. pyogenes and their capacity to control infection was compared. Aged mice showed faster progression of disease, earlier morbidity, and increased mortality when compared with young animals. Since macrophages are critical for host defence against S. pyogenes, we investigated whether susceptibility of aged mice may be due to an age-associated decline in the functionality of these cells. Our results showed that macrophages from aged mice were as capable as those from young animals to uptake and kill S. pyogenes, but the number of resident tissue macrophages was significantly reduced in the aged host. Treatment of aged mice with macrophage colony-stimulating factor (M-CSF) significantly increased the number of resident macrophages and improved their response to infection. Our results indicate that treatment with M-CSF can restore, at least in part, the mechanisms affected by immunosenescence and enhance the natural resistance of aged mice to infection with S. pyogenes. PMID- 20020514 TI - Effect of pomegranate juice on Angiotensin II-induced hypertension in diabetic Wistar rats. AB - Acute subcutaneous administration of Angiotensin II (Ang II) causes a rise in blood pressure in diabetic Wistar rats. Diabetes was induced using streptozotocin (70 mg/kg, i.v.). Chronic administration of pomegranate juice (PJ) extract (100 mg/kg and 300 mg/kg; p.o. for 4 weeks) obtained from Punica granatum (punicaceae) fruits reduced the mean arterial blood pressure and vascular reactivity changes to various catecholamines and also reversed the biochemical changes induced by diabetes and Ang II. PJ treatment also caused a significant decrease in levels of thiobarbituric acid reactive substances (TBARS) in kidney and pancreas while activities of enzymes superoxide dismutase (SOD), catalase (CAT), and glutathione reductase (GSH) showed significant elevation. The cumulative concentration response curve (CCRC) of Ang II was shifted towards right in rats treated with PJ using isolated strip of ascending colon. In histopathological examination, PJ treatment prevented the tubular degenerative changes induced by diabetes. The results suggest that the PJ extract could prevent the development of high blood pressure induced by Ang II in diabetic rats probably by combating the oxidative stress induced by diabetes and Ang II and by inhibiting ACE activity. In conclusion, PJ has antihypertensive action in Ang II diabetic model. PMID- 20020515 TI - Highly sensitive indicator-free impedance sensing of DNA hybridization based on poly(m-aminobenzenesulfonic acid)/TiO2 nanosheet membranes with pulse potentiostatic method preparation. AB - A direct electrochemical detection procedure for DNA hybridization by using the electrochemical signal changes of conductive poly(m-aminobenzenesulfonic) acid (PABSA)/TiO(2) nanosheet membranes, which were electropolymerized by using the pulse potentiostatic method, is reported. Due to the unique properties of TiO(2) nanoparticles, m-aminobenzenesulfonic acid monomers tend to be adsorbed around the particles, and the electropolymerization efficiency is greatly improved. The combination of TiO(2) nanoparticles and PABSA resulted in a nanocomposite membrane with unique and novel nanosheet morphology that provides more activation sites and enhances the surface electron-transfer rate. These characteristics were propitious for the magnification of PABSA electrochemical signals and the direct detection of DNA hybridization. Owing to the presence of abundant sulfonic acid groups, PABSA could overcome the drawbacks of polyaniline and be used to detect bioanalytes at physiological pH. DNA probes could be covalently attached to the sulfonic groups through the amines of DNA sequences by using an acyl chloride cross-linking reaction. After immobilization of probe DNA, the electrochemical impedance value increased significantly compared to that of PABSA/TiO(2) nanosheet membranes, and then decreased dramatically after the hybridization reaction of the probe DNA with the complementary DNA sequence compared to that of the probe-immobilized electrode. Electrochemical impedance spectroscopy was adopted for indicator-free DNA biosensing, which had an eminent ability for the recognition between double-base mismatched sequences or non-complementary DNA sequences and complementary DNA sequences. A gene fragment, which is related to one of the screening genes for the transgenically modified plants, the cauliflower mosaic virus 35S gene was satisfactorily detected. This is the first report for the indicator-free impedance DNA hybridization detection by using PABSA/TiO(2) membranes under neutral conditions. PMID- 20020516 TI - Amino acid based low-molecular-weight ionogels as efficient dye-adsorbing agents and templates for the synthesis of TiO(2) nanoparticles. AB - The gelation of ionic liquids is attracting significant attention because of its large spectrum of applications across different disciplines. These 'green solvents' have been the solution to a number of common problems due to their eco friendly features. To expand their applications, the gelation of ionic liquids has been achieved by using amino acid-based low-molecular-weight compounds. Variation of individual segments in the molecular skeleton of the gelators, which comprise the amino acid and the protecting groups at the N and C termini, led to an understanding of the structure-property correlation of the ionogelation process. An aromatic ring containing amino acid-based molecules protected with a phenyl or cyclohexyl group at the N terminus were efficient in the gelation of ionic liquids. In the case of aliphatic amino acids, gelation was more prominent with a phenyl group as the N-terminal protecting agent. The probable factors responsible for this supramolecular association of the gelators in ionic liquids have been studied with the help of field-emission SEM, (1)H NMR, FTIR, and luminescence studies. It is the hydrophilic-lipophilic balance that needs to be optimized for a molecule to induce gelation of the green solvents. Interestingly, to maximize the benefits from using these green solvents, these ionogels have been employed as templates for the synthesis of uniform-sized TiO(2) nanoparticles (25-30 nm). Furthermore, as a complement to their applications, ionogels serve as efficient adsorbents of both cationic and anionic dyes and were distinctly better relative to their organogel counterparts. PMID- 20020517 TI - Tuning the functional properties of metal complexes containing polytopic heteroaromatic nitrogen ligands. AB - The preparation, characterization, and optimization of the functional properties of mono- and polynuclear coordination complexes containing heteroaromatic nitrogen ligands are discussed here, taking the advantage of numerous studies performed in our laboratories on exploring a variety of different metal ions and polytopic ligands. We highlight how very minor changes in connectivity, composition, and polarity of the molecular entities employed in the self-assembly steps may significantly affect the structural, thermal, sorptive, magnetic, and mesomorphic behavior of the resulting materials. Examples from three different classes are included: 1) pyrazolate-based polynuclear coordination compounds, 2) homoleptic and heteroleptic coordination polymers, and 3) 2,2'-bipyridine metal based liquid crystals. PMID- 20020518 TI - Boron-containing protoporphyrin IX derivatives and their modification for boron neutron capture therapy: synthesis, characterization, and comparative in vitro toxicity evaluation. AB - A novel series of boronated porphyrins for potential use in boron neutron capture therapy (BNCT) and photodynamic therapy (PDT) for tumor suppression is described. Protoporphyrin IX {i.e., bis(alpha-methyl-beta-pentylethylether)protoporphyrin IX, and bis(alpha-methyl-beta-dodecanylethylether)protoporphyrin IX} bearing polyhedral borane anions (B(12)H(11)SH(2-), B(12)H(11)NH(3) (-), or B(12)H(11)OH(2-)) were synthesized with reasonable yields. Modification of the protoporphyrin IX structure was achieved by variation of the lengths of the alkyl chains (pentyl and dodecanyl) attached through ether linkages to the former vinyl groups. The goal of this modification was to develop boronated porphyrins with chemical and physical properties that differed from those of protoporphyrin IX. Performance of an MTT assay with each derivative revealed that the synthesized boronated porphyrins showed low cytotoxicities in a variety of cancer cells. Of these compounds, B(12)H(11)NH(2) (2-)-conjugated porphyrin induced a significant increase in the level of boron accumulation and PDT efficacy against HeLa cells. PMID- 20020520 TI - Carbon nanotubes fed on "carbs": coating of single-walled carbon nanotubes by dextran sulfate. AB - A facile and easily reproducible technique for assembling biohybrid nanoparticles is a core feature that is highly desired for biomedical applications, considering the nature and limited lifespan of the biopolymers used. Here we show a simple and effective method to enfold single-walled carbon nanotubes (SWNTs) using an anionic polysaccharide, dextran sulfate. After their interactions, SWNTs were rendered dispersible in aqueous solution and were shortened and unbundled to their basic dimension. Atomic force microscopy analysis was extensively employed to elucidate the mechanism of their interfacing. This biohybrid nanoparticle holds promise for biological and biomedical applications due to the synergistic unique properties of SWNTs and dextran sulfate. PMID- 20020519 TI - Improving gene delivery efficiency of bioreducible poly(amidoamine)s via grafting with dendritic poly(amidoamine)s. AB - Dendritic poly(amidoamine)s (PAMAM)s were introduced into the side chains of disulfide-containing poly(amidoamine)s via repetitive Michael addition and amidation. The bioreducible poly(amidoamine)s grafted with dendritic polyamidoamines showed high buffer capacity, low cytotoxicity and strong DNA binding ability at low N/P ratio. They were able to condense DNA into small sized polycation/DNA complexes, which degraded and released the incorporated DNA under reductive conditions. In comparison to the original disulfide-containing poly(amidoamine) with aminoethyl side chain, the grafting of the bioreducible poly(amidoamine) with dendrimer greatly improved the transfection efficiencies of 293T and HeLa cells with foreign DNA at various N/P ratios. The structure-gene delivery property relations of dendrimer-grafted polycations will provide valuable insight into the design of highly efficient and less toxic polycationic gene carriers. PMID- 20020521 TI - Brain salvage for cardiac cerebral embolism following myocardial infarction. AB - Acute ischemic stroke due to thromboembolism from the left ventricle is a serious complication following myocardial infarction (MI). Despite the attention given to acute stroke intervention, peri-MI ischemic stroke is an underappreciated condition. Here, we describe a case of acute cardiac thromboembolic occlusion of the middle cerebral artery with preceding MI in which brain salvage was achieved by balloon angioplasty. PMID- 20020522 TI - Visual inspection versus quantitative flow cytometry to detect aberrant CD2 expression in malignant T cells. AB - BACKGROUND: Abnormal levels of T-cell antigen expression occur in T-cell neoplasia. We examined CD2 expression in malignant and normal T cells to determine if the level of CD2 expression differed significantly and if quantitation assisted in detecting this difference. METHOD: Flow cytometric immunophenotypic (FCI) evaluation was performed on specimens from 36 patients with mature T-cell neoplasia. Abnormal T cells were identified based upon the abnormal FCI and morphology. Levels of CD2 expression were quantitated using 1:1 PE conjugates of anti-CD2 and QuantiBRITE bead standards to calculate the antibodies bound per cell (ABC). The efficacy of ABC measurement versus simple examination of dots plots was compared. RESULTS: Abnormal levels of CD2 expression were frequently observed in mature T-cell malignancies. The CD2 ABC values were highly sensitive in detecting differences between malignant and normal T cells (P = 0.0028). In most cases (24/32 specimens, 75%), CD2 ABCs differed by >20%. CD2 ABCs had high variability in normal T cells. CONCLUSIONS: CD2 expression by malignant T cells differed significantly from that of normal T cells by CD2 ABC quantitation. The high variability in normal T-cell CD2 ABCs limited the determination of normal reference ranges and, thus, its utility in the diagnosis of T-cell neoplasia. However, examination of CD2 can help in detection of tumor cells when residual normal T cells are present for comparison. Moreover, the increased sensitivity of CD2 quantitation is valuable in confirming FCI cases where abnormalities in CD2 expression are difficult to appreciate by visual inspection alone. PMID- 20020523 TI - Molecular cloning and characterization of the germ cell-related nuclear orphan receptor in chickens. AB - Nuclear receptor subfamily 6, group A, member 1 (NR6A1), also known as germ cell nuclear factor/retinoid receptor-related testis-associated receptor and neuronal cell nuclear factor, is a member of the nuclear orphan receptor superfamily. NR6A1 has been cloned in various species including humans and mice, but it has been scarcely investigated in avian species. In the present study, we cloned the chicken NR6A1 (cNR6A1) from a testis cDNA library. The cloned cNR6A1 sequence was mapped to chromosome 17 and contained an open reading frame of 1.4 kb encoding 445 amino acids. Multiple alignment analysis of the cNR6A1 protein-coding sequence with NR6A1s from humans, mice, boars, rats, zebrafish, and Xenopus showed high degrees of homology, 89%, 90%, 89%, 88%, 83%, and 87%, respectively. Using RNA interference, changes in the expression of pluripotency-, germ cell-, and differentiation-related key genes by silencing of cNR6A1 were validated in chicken blastoderm-derived embryonic stem cells. Among those genes, the relative expression levels of POU5F1, CRIPTO, DAZL, DDX4, BMP15, GSC, and SOX7 changed significantly compared to the control group. We also confirmed that the activity of alkaline phosphatase, known as a pluripotency marker, was maintained by cNR6A1 gene silencing in chicken blastodermal cells. Collectively, our data suggest that cNR6A1 may play an important role during chicken embryonic development and differentiation. PMID- 20020525 TI - A novel prodrug strategy for beta-dicarbonyl carbon acids: syntheses and evaluation of the physicochemical characteristics of C-phosphoryloxymethyl (POM) and phosphoryloxymethyloxymethyl (POMOM) prodrug derivatives. AB - The C-phosphoryloxymethyl (POM) and phosphoryloxymethyloxymethyl (POMOM) prodrugs resulting from derivatization at the reactive alpha-carbon of beta-dicarbonyl carbon acid drugs represent a unique approach for improving their chemical stability and aqueous solubility. This work evaluates the physicochemical and in vitro enzymatic bioconversion lability of selected prodrugs of phenylbutazone and phenindione. The POM and POMOM prodrug derivatives of phenylbutazone are highly water soluble (>or=250 mg/mL), chemically stable with projected shelf-lives of 4.5 years (pH 3.5, 25 degrees C) and 1.1 years (pH 6.0, 25 degrees C), respectively. Interestingly, both prodrug derivatives do not display a pH dependency typical of many phosphate monoesters, although the similarities of their apparent thermodynamic activation parameters indicate a hydrolysis mechanism similar to other phosphates. These prodrugs undergo alkaline phosphatases catalyzed bioconversion to their respective carbon acids with an expected faster rate exhibited by the POMOM derivatives. Additionally, in marked contrast to the oxidative instability of phenindione, its POM prodrug is stable. The results from these studies reaffirm the rationale of transiently "masking" the reactive alpha-carbon/proton bond by covalently incorporating a POM or POMOM promoiety. This prodrug strategy presents a twofold advantage, enhancement of aqueous solubility and prevention of oxidative instability, two intrinsic formulation limitations found for beta-dicarbonyl carbon acid drugs. PMID- 20020526 TI - Effect of altered AGP plasma binding on heart rate changes by S(-)-propranolol in rats using mechanism-based estimations of in vivo receptor affinity (K(B,vivo)). AB - In contrast to the impact of plasma protein binding on pharmacokinetics, no quantitative in vivo information is available on its impact on pharmacodynamics. The pharmacokinetic-pharmacodynamic relationship of the model drug S(-) propranolol was evaluated using mechanism-based estimations of in vivo receptor affinity (K(B,vivo)), under conditions of altered plasma protein binding resulting from different levels of alpha-1-acid glycoprotein (AGP). Male Wistar Kyoto rats with isoprenaline-induced tachycardia received an intravenous infusion of S(-)-propranolol, on postsurgery day 2 (n = 7) and day 7 (n = 8) with elevated and normal plasma protein binding, respectively. Serial blood samples were taken in parallel to heart rate measurements. AGP concentrations at 2 and 7 days postsurgery were 708 +/- 274 and 176 +/- 111 microg/mL (mean +/- SE), respectively. Using nonlinear mixed effects modeling, AGP concentration was a covariate for intercompartmental clearance for the third compartment of the pharmacokinetic model of S(-)-propranolol. Individual values of AGP concentrations ranged between 110 and 1150 microg/mL, and were associated with K(B,vivo) values of S(-)-propranolol from 7.0 to 30 nM. Using the K(B,vivo) for S(-)-propranolol with correction for average values for normal and elevated plasma protein binding, nearly identical values were found. This confirms, strictly quantitative, earlier indications that plasma protein binding restricts the pharmacodynamics of S(-)-propranolol. PMID- 20020528 TI - Generation of functional hepatocytes from mouse induced pluripotent stem cells. AB - Induced pluripotent stem cells are derived from somatic cells by forced expression of several transcriptional factors. Induced pluripotent stem cells resemble embryonic stem cells in many aspects, such as the expression of certain stem cell markers, chromatin methylation patterns, embryoid body formation and teratoma formation. Therefore, induced pluripotent stem cells provide a powerful tool for study of developmental biology and unlimited resources for transplantation therapy. Here we reported the successful induction of mouse induced pluripotent stem cells and a simple and efficient process for generation of functional hepatocytes from mouse induced pluripotent stem cells by sequential addition of inducing factors. These induced pluripotent stem cell-derived hepatocytes, just as mouse embryonic stem cell-derived hepatocytes, expressed hepatic lineage markers including CK7, CK8, CK18, CK19, alpha-fetoprotein, albumin, Cyp7a1, and exhibited functional hepatic characteristics, including glycogen storage, indocyanine green (ICG) uptake and release, low-density lipoprotein (LDL) uptake and urea secretion. Although we observed some variations in the efficiency of hepatic differentiation between induced pluripotent stem cells and common mouse embryonic stem cell lines, our results indicate that mouse induced pluripotent stem cells can efficiently differentiate into functional hepatocytes in vitro, which may be helpful for the study of liver development and regenerative medicine. PMID- 20020529 TI - Bayes analysis provides evidence of pathogenicity for the BRCA1 c.135-1G>T (IVS3 1) and BRCA2 c.7977-1G>C (IVS17-1) variants displaying in vitro splicing results of equivocal clinical significance. AB - Although in vitro splicing assays can provide useful information about the clinical interpretation of sequence variants in high-risk cancer genes such as BRCA1 and BRCA2, results can sometimes be difficult to interpret. The BRCA1 c.135 1G>T (IVS3-1G>T) variant has been shown to give rise to an in-frame deletion of exon 5 (BRCA1 c.135_212del) that is predicted to encode 26 amino acids. BRCA2 c.7977-1G>C (IVS17-1G>C) was shown to increase the expression of two naturally occurring transcripts that contain frameshifts (BRCA2, c.7977_8311del (exon 18 deletion); BRCA2, c.7806_8331del (exon 17&18 deletion)). In this study we conducted multifactorial likelihood analysis to evaluate the clinical significance of these two variants, including assessing variant segregation in families by Bayes analysis, and breast tumor pathology features suggestive of positive mutation status. Multifactorial analysis provided strong evidence for causality for both of these variants. The Bayes scores from a single family with BRCA1 c.135-1G>T was 9528:1, and incorporation of pathology features gave an overall likelihood of causality of 28108:1. The Bayes scores from five informative families with BRCA2 c.7977-1G>C was 47401:1, and the combined Bayes pathology odds of causality was 29389:1. Multifactorial likelihood analysis indicates that the BRCA1 c.135-1G>T and BRCA2 c.7977-1G>C variants are disease associated mutations which should be managed clinically in the same fashion as classical truncating mutations. PMID- 20020527 TI - Defining the heterogeneity of skeletal muscle-derived side and main population cells isolated immediately ex vivo. AB - Myoblast transfer therapy for Duchenne muscular dystrophy (DMD) largely fails due to cell death and inability of transplanted cells to engraft in diseased muscles. One method attempting to enrich for cell subpopulations is the Hoechst 33342 dye exclusion assay, yielding a side population (SP) thought to be progenitor enriched and a main population (MP). However, in vitro and transplant studies yielded inconsistent results relative to downstream progeny. Cell surface markers expressed by skeletal muscle-derived MP and SP cells have not been fully characterized directly ex vivo. Using flow cytometry, MP and SP cells were characterized based on their expression of several well-accepted progenitor cell antigens. Both the MP and SP populations are heterogeneous and overlapping in the cells they contain. The percentages of cells in each population vary with species and specific muscle examined. MP and SP populations contain both satellite and multipotent progenitor cells, based on expression of CD34, Sca-1, Pax7, and M cadherin. Thus, isolation using this procedure cannot be used to predict downstream differentiation outcomes, and explains the conflicting literature on these cells. Hoechst dye also results in significant mortality of sorted cells. As defined subpopulations are easily obtained using flow cytometry, sorting immediately ex vivo based on accepted myogenic precursor cell markers will yield superior results in terms of cell homogeneity for transplantation therapy. PMID- 20020530 TI - NKX2-1 mutations leading to surfactant protein promoter dysregulation cause interstitial lung disease in "Brain-Lung-Thyroid Syndrome". AB - NKX2-1 (NK2 homeobox 1) is a critical regulator of transcription for the surfactant protein (SP)-B and -C genes (SFTPB and SFTPC, respectively). We identified and functionally characterized two new de novo NKX2-1 mutations c.493C>T (p.R165W) and c.786_787del2 (p.L263fs) in infants with closely similar severe interstitial lung disease (ILD), hypotonia, and congenital hypothyroidism. Functional analyses using A549 and HeLa cells revealed that NKX2-1-p.L263fs induced neither SFTPB nor SFTPC promoter activation and had a dominant negative effect on wild-type (WT) NKX2-1. In contrast,NKX2-1-p.R165W activated SFTPC, to a significantly greater extent than did WTNKX2-1, while SFTPB activation was only significantly reduced in HeLa cells. In accordance with our in vitro data, we found decreased amounts of SP-B and SP-C by western blot in bronchoalveolar lavage fluid (patient with p.L263fs) and features of altered surfactant protein metabolism on lung histology (patient with NKX2-1-p.R165W). In conclusion, ILD in patients with NKX2-1 mutations was associated with altered surfactant protein metabolism, and both gain and loss of function of the mutated NKX2-1 genes on surfactant protein promoters were associated with ILD in "Brain-Lung-Thyroid syndrome". PMID- 20020532 TI - A haplotype of the catalase gene confers an increased risk of essential hypertension in Chinese Han. AB - Our previous study in an isolated population showed an association between a genetic variant in the catalase gene (CAT) and essential hypertension (EH). This study indicates that three variants in the promoter and 5'-UTR region of CAT are predominant in Chinese Han, and they form two major haplotypes. A case-control study showed that the CATH2 haplotype confers susceptibility to EH (Pgenotype=0.0017, and Pallilc=0.00078). Subjects bearing CATH1/CATH2 and CATH2/CATH2 genotypes demonstrated a higher susceptibility to EH than CATH1/CATH1 homozygotes, with odds ratios of 1.474 and 1.625, respectively. Also, CATH1/CATH1 individuals had a later-onset age (P=0.015). Expression analysis using luciferase reporter vectors indicated that the CATH1 haplotype showed a lower transcriptional activity than the haplotype CATH2 (P<0.05 in all four cell lines), and we observed similar results in the endogenous allelic expression ratios of CATH1/CATH2 in cell lines. In contrast, most CATH1 haplotypes showed a higher transcription level than CATH2 haplotypes (10 out of 11 or 90.9%) in blood from normal individuals (P<0.01). We therefore hypothesize that CATH1 and CATH2 may play alternating roles at different level of oxidative stress. PMID- 20020531 TI - A thorough assessment of benign genetic variability in GRN and MAPT. AB - Mutations in APP, PSEN1, MAPTand GRNare the most common genetic causes of dementia. The previous miss-assignment of pathogenicity to benign variants in these genes stresses the importance of discerning between disease causing mutations and benign variants with no pathogenic effect on the function of the respective protein. In this study we sequenced GRNand MAPTin 282 samples from the Centre d'Etude du Polymorphisme Humain - Human Genome Diversity Cell Line Panel, in order to identify benign variants that could otherwise be mistaken for pathogenic mutations. We found sixteen different non-synonymous changes, eleven of which are novel variants. PMID- 20020533 TI - Evidence for genetic heterogeneity in D-2-hydroxyglutaric aciduria. AB - We performed molecular, enzyme, and metabolic studies in 50 patients with D-2 hydroxyglutaric aciduria (D-2-HGA) who accumulated D-2-hydroxyglutarate (D-2-HG) in physiological fluids. Presumed pathogenic mutations were detected in 24 of 50 patients in the D-2-hydroxyglutarate dehydrogenase (D2HGDH) gene, which encodes D 2-hydroxyglutarate dehydrogenase (D-2-HGDH). Enzyme assay of D-2-HGDH confirmed that all patients with mutations had impaired enzyme activity, whereas patients with D-2-HGA whose enzyme activity was normal did not have mutations. Significantly lower D-2-HG concentrations in body fluids were observed in mutation-positive D-2-HGA patients than in mutation-negative patients. These results imply that multiple genetic loci may be associated with hyperexcretion of D-2-HG. Accordingly, we suggest a new classification: D-2-HGA Type I associates with D-2-HGDH deficiency, whereas idiopathic D-2-HGA manifests with normal D-2 HGDH activity and higher D-2-HG levels in body fluids compared with Type I patients. It remains possible that several classifications for idiopathic D-2-HGA patients with diverse genetic loci will be revealed in future studies. PMID- 20020534 TI - AlphaIIbbeta3 integrin: new allelic variants in Glanzmann thrombasthenia, effects on ITGA2B and ITGB3 mRNA splicing, expression, and structure-function. AB - Glanzmann thrombasthenia (GT) is an autosomal recessive inherited bleeding disorder characterized by an impaired platelet aggregation due to defects in integrin alphaIIbbeta3 (ITGA2B, ITGB3), a fibrinogen receptor. Mutations from 24 GT patients and two carriers of various origins, Caucasian, North-African and Asian were characterized. Promoter and exon sequences of alphaIIb and beta3 genes were amplified and directly sequenced. Among 29 identified mutations, 17 new allelic variants resulting from nonsense, missense and deletion/insertion mutations were described. RNA alterations were evaluated by using Web servers. The alphaIIb p.S926L, p.V903F, and beta3 p.C38Y, p.M118R, p.G221D substitutions prevented complex expression at the surface of COS-7 cells by altering the alphaIIb or the beta3 subunit structure. As shown by free energy analyses applied on the resolved structure of alphaIIbbeta3 and structural modeling of the mutant, the p.K253M substitution of beta3 helped to define a key role of the K253 in the interaction of the alphaIIb beta-propeller and the beta3 beta-I domains. finally, the alphaIIb p.Q595H substitution allowed cell surface expression of the complex but its corresponding c.2800G>T mutation is predicted to alter normal RNA splicing. In conclusion, our study yielded the discovery of 17 new GT allelic variants, revealed the key role of K253 of alphaIIb for the alphaIIbbeta3 complex formation and provides an additional example of an apparently missense mutation causing a splicing defect. PMID- 20020535 TI - A cell-free assay for the functional analysis of variants of the mismatch repair protein MLH1. AB - The hereditary colon and endometrium cancer predisposition Lynch Syndrome (also called HNPCC) is caused by a germ-line mutation in one of the DNA mismatch repair (MMR) genes. A significant fraction of the gene alterations detected in suspected Lynch Syndrome patients is comprised of amino acid substitutions. The relevance for cancer risk of these variants is difficult to assess, as currently no time- and cost-effective, validated, and widely applicable functional assays for the measurement of MMR activity are available. Here we describe a rapid, cell-free, and easily quantifiable MMR activity assay for the diagnostic assessment of variants of the MLH1 MMR protein. This assay allows the parallel generation and functional analysis of a series of variants of the MLH1 protein in vitro using readily available, or preprepared, reagents. Using this assay we have tested 26 MLH1 variants and of these, 15 had lost activity. These results are in concordance with those obtained from first-generation assays and with in silico and pathology data. After its multifocal technical and clinical validation this assay could have great impact for the diagnosis and counseling of carriers of an MLH1 variant and their relatives. PMID- 20020536 TI - Pathogenic mutations cause rapid degradation of lysosomal storage disease-related membrane protein CLN6. AB - One variant form of late infantile neuronal ceroid lipofuscinosis is an autosomal recessive inherited neurodegenerative lysosomal storage disorder caused by mutations in the CLN6gene. The function of the polytopic CLN6 membrane protein localized in the endoplasmic reticulum is unknown. Here we report on expression studies of three mutations (c.368G>A, c.460-462delATC, c.316insC) found in CLN6 patients predicted to affect transmembrane domain 3 (p.Gly123Asp), cytoplasmic loop 2 (p.Ile154del) or result in a truncated membrane protein (p.Arg106ProfsX26), respectively. The rate of synthesis and the stability of the mutant CLN6 proteins are reduced in a mutation-dependent manner. None of the mutations prevented the dimerization of the CLN6 polypeptides. The particularly rapid degradation of the p.Arg106ProfsX26 mutant which is identical with the mutation in the murine orthologue Cln6 gene in the nclf mouse model of the disease, can be strongly inhibited by proteasomal and partially by lysosomal protease inhibitors. Both degradative pathways seem to be sufficient to prevent the accumulation/aggregation of the mutant CLN6 polypeptides in the endoplasmic reticulum. PMID- 20020537 TI - A pharmacogenetic study of escitalopram in autism spectrum disorders. AB - OBJECTIVE: To determine the effect of serotonin transporter polymorphism promoter region (5-HTTPLR) genotypic variation (low, intermediate, and high expression groups) on response to escitalopram treatment of children and adolescents with autism spectrum disorders (ASDs). METHOD: The study used a forced titration, open label design, with genotype blind until study completion. Participants were children and adolescents aged 4-17 years of age with a confirmed ASD (autistic disorder, Asperger's disorder, or pervasive developmental disorder, not otherwise specified). RESULTS: There was an interaction between genotype group and time on the Aberrant Behavior Checklist (ABC) Irritability Subscale (primary outcome variable) (linear maximum marginal likelihood estimation=-4.84, Z=-2.89, SE=1.67, P=0.004). Examination of baseline to last visit revealed that a genotype grouping based on a previous study of platelet 5-HT uptake revealed less response in the genotype group that had S/S genotype for 5-HTTLPR and did not have a diplotype in intron 1 previously shown to be associated with increased platelet 5-HT uptake. CONCLUSION: This genotype-blind, prospective pharmacogenetic study found the group of subjects with associated with the lowest platelet 5-HT uptake from previous study had the smallest reduction in ABC-Irritability scores after open label treatment with escitalopram. Replication is necessary to confirm these findings. PMID- 20020538 TI - Microglial response to light-induced photoreceptor degeneration in the mouse retina. AB - The microglial response elicited by degeneration of retinal photoreceptor cells was characterized in BALB/c mice exposed to bright light for 7 hours and then kept in complete darkness for survival times ranging from 0 hours to 10 days. Photodegeneration resulted in extensive cell death in the retina, mainly in the outer nuclear layer (ONL), where the photoreceptor nuclei are located. Specific immunolabeling of microglial cells with anti-CD11b, anti-CD45, anti-F4/80, anti SRA, and anti-CD68 antibodies revealed that microglial cells were activated in light-exposed retinas. They migrated to the ONL, changed their morphology, becoming rounded cells with short and thick processes, and, finally, showed immunophenotypic changes. Specifically, retinal microglia began to strongly express antigens recognized by anti-CD11b, anti-CD45, and anti-F4/80, coincident with cell degeneration. In contrast, upregulation of the antigen recognized by anti-SRA was not detected by immunocytochemistry until 6 hours after light exposure. Differences were also observed at 10 days after light exposure: CD11b, CD45, and F4/80 continued to be strongly expressed in retinal microglia, whereas the expression of CD68 and SRA had decreased to near-normal values. Therefore, microglia did not return to their original state after photodegeneration and continued to show a degree of activation. The accumulation of activated microglial cells in affected regions simultaneously with photoreceptor degeneration suggests that they play some role in photodegeneration. PMID- 20020539 TI - Localization of the calcium-binding protein secretagogin in cone bipolar cells of the mammalian retina. AB - Secretagogin, a recently cloned member of the EF-hand family of calcium binding proteins, was localized in the mouse, rat, and rabbit retina using immunofluorescence immunohistochemistry. Secretagogin is expressed in subpopulations of ON and OFF cone bipolar cells; however, no immunoreactivity was observed in rod bipolar cells in any of these species. Using subtype-specific markers and mice expressing green fluorescent protein (GFP) within specific cell classes, we found that secretagogin is expressed in Types 2, 3, 4, 5, 6 and possibly Type 8 cone bipolar cells in the mouse retina. The expression pattern in the rat retina differs slightly with expression in cone bipolar cell Types 2, 5, 6, 7, and 8. Evaluation of secretagogin in the developing mouse retina revealed expression as early as postnatal day 6, with OFF cone bipolar cells showing secretagogin expression prior to the ON cone bipolar cells. Secretagogin is a useful marker of cone bipolar cells for studying alterations in bipolar cell morphology during development and degeneration. Further work will be necessary to elucidate the functional role of this protein in bipolar cells. PMID- 20020542 TI - Heart health after breast cancer. PMID- 20020541 TI - Differentiation of papillae and rostral sensory neurons in the larva of the ascidian Botryllus schlosseri (Tunicata). AB - During the metamorphosis of tunicate ascidians, the swimming larva uses its three anterior papillae to detect the substrate for settlement, reabsorbs its chordate like tail, and becomes a sessile oozooid. In view of the crucial role played by the anterior structures and their nerve relations, we applied electron microscopy and immunocytochemistry to study the larva of the colonial ascidian Botryllus schlosseri, following differentiation of the anterior epidermis during late embryogenesis, the larval stage, and the onset of metamorphosis. Rudiments of the papillae appear in the early tail-bud stage as ectodermic protrusions, the apexes of which differentiate into central and peripheral bipolar neurons. Axons fasciculate into two nerves direct to the brain. Distally, the long, rod-like dendritic terminations extend during the larval stage, becoming exposed to sea water. After the larva selects and adheres to the substrate, these neurons retract and regress. Adjacent to the papillae, other scattered neurons insinuate dendrites into the tunic and form the net of rostral trunk epidermal neurons (RTENs) which fasciculate together with the papillary neurons. Our data indicate that the papillae are simple and coniform, the papillary neurons are mechanoreceptors, and the RTENs are chemoreceptors. The interpapillary epidermal area, by means of an apocrine secretion, provides sticky material for temporary adhesion of the larva to the substrate. PMID- 20020540 TI - Distribution of androgen receptor mRNA expression in vocal, auditory, and neuroendocrine circuits in a teleost fish. AB - Across all major vertebrate groups, androgen receptors (ARs) have been identified in neural circuits that shape reproductive-related behaviors, including vocalization. The vocal control network of teleost fishes presents an archetypal example of how a vertebrate nervous system produces social, context-dependent sounds. We cloned a partial cDNA of AR that was used to generate specific probes to localize AR expression throughout the central nervous system of the vocal plainfin midshipman fish (Porichthys notatus). In the forebrain, AR mRNA is abundant in proposed homologs of the mammalian striatum and amygdala, and in anterior and posterior parvocellular and magnocellular nuclei of the preoptic area, nucleus preglomerulosus, and posterior, ventral and anterior tuberal nuclei of the hypothalamus. Many of these nuclei are part of the known vocal and auditory circuitry in midshipman. The midbrain periaqueductal gray, an essential link between forebrain and hindbrain vocal circuitry, and the lateral line recipient nucleus medialis in the rostral hindbrain also express abundant AR mRNA. In the caudal hindbrain-spinal vocal circuit, high AR mRNA is found in the vocal prepacemaker nucleus and along the dorsal periphery of the vocal motor nucleus congruent with the known pattern of expression of aromatase-containing glial cells. Additionally, abundant AR mRNA expression is shown for the first time in the inner ear of a vertebrate. The distribution of AR mRNA strongly supports the role of androgens as modulators of behaviorally defined vocal, auditory, and neuroendocrine circuits in teleost fish and vertebrates in general. PMID- 20020543 TI - Celiac disease: a concern even later in life. PMID- 20020544 TI - Consultation corner. Confronting controversies over antidepressants. PMID- 20020545 TI - For COPD: get thee to pulmonary rehab. PMID- 20020546 TI - If my doctor's office doesn't call with my test results, can I assume that everything is normal? PMID- 20020547 TI - Prevalence of mental illness in older Australians. PMID- 20020548 TI - Lycanthropy in alcohol intoxication. PMID- 20020549 TI - [Perioperative management of chronic treatment and medical devices. Preamble]. PMID- 20020550 TI - [Perioperative management of chronic treatment and medical devices. Cardiovascular pathologies]. PMID- 20020551 TI - [Perioperative management of chronic treatment and medical devices: cardiovascular pathologies]. PMID- 20020552 TI - [Perioperative management of chronic treatment and medical devices: anti infectious, immunosuppressive]. PMID- 20020553 TI - Common cosmetic procedures. PMID- 20020554 TI - Coronary artery disease and the use of stents. PMID- 20020555 TI - Von Willebrand Disease: a bleeding disorder. PMID- 20020556 TI - [Palace and garden: the relationship between nature and art]. PMID- 20020557 TI - [Watteau, Marivaux, and fetes galantes: in other words, what can happen in the garden?]. PMID- 20020558 TI - [Woman's fate as described in the 17th- and 18th-century Ottoman books of complaints]. PMID- 20020559 TI - [The aesthetic influence of Andre Lenotre on designs for the surroundings of Polish residences at the end of the 17th century and during the first half of the 18th century]. PMID- 20020560 TI - [A treatise on gardens by Tomasz Antoni Zamoyski, ca. 1750]. PMID- 20020561 TI - [The project for an extension of the Wilanow gardens, ca. 1740]. PMID- 20020562 TI - Agomelatine: new drug. Adverse effects and no proven efficacy. AB - (1) When an antidepressant is considered a necessary addition to psychological support in treating patients with depression, the first-line drug is a tricyclic such as clomipramine or a selective serotonin reuptake inhibitor (SSRI) such as paroxetine; (2) Agomelatine, a melatonin receptor agonist, is approved in the European Union for the treatment of depression; (3) Available evaluation does not include any clinical trials designed to compare the efficacy of agomelatine with that of a tricyclic or a selective serotonin reuptake inhibitor. Most data come from 7 placebo-controlled trials; (4) Agomelatine (25 mg/day) was statistically more effective (on a rating scale) than placebo in only 3 of these 7 trials. The clinical relevance of the score improvements is questionable. No data are available on the cure rate or on suicide prevention; (5) In one trial, increasing the daily dose from 25 mg to 50 mg provided no supplementary benefit; (6) A trial in 367 patients failed to show that agomelatine was any more effective than placebo in preventing new depressive episodes (29% after one year). In another trial including 339 patients, the relapse rate was statistically lower after 6 months on agomelatine (20.6%) than on placebo (41.4%); (7) Very high doeses of agomelatine are oncogenic in animals. The risk in humans is not known. Dizziness, gastrointestinal and cutaneous disorders have been observed. Agomelatine is probably hepatotoxic; (8) In summary, agomelatine has unproven efficacy and poorly documented adverse effects. It is better to continue to use older antidepressants such as tricyclics or serotonin reuptake inhibitors. PMID- 20020564 TI - Drug interactions with new and investigational antiretrovirals. PMID- 20020563 TI - [Ileus of uncommon cause--important and weighty facts]. AB - HISTORY AND ADMISSION FINDINGS: A 36-year-old female patient was admitted for abdominal pain and recurrent vomiting since five days. One year before this episode, a gastric balloon had been implanted. Physical examination revealed tenderness in the left and right lower abdomen. INVESTIGATIONS: Abdominal ultrasound showed marked impairment of intestinal motility and plain abdominal x ray revealed signs of small intestine ileus. On abdominal and pelvic computed tomography the gastric balloon was dislocated to the terminal ileum. DIAGNOSIS, TREATMENT AND COURSE: Dislocation of the gastric balloon resulted in occlusion of the ileum and consequently in mechanical small intestine ileus. After an unsuccessful endoscopic attempt, the balloon was removed during laparatomy. After an initially protracted postinterventional course with impairment of gastrointestinal motility, gradual return to solid food was without any difficulty in the following course. CONCLUSION: Due to the pandemic increase of the obesity incidence and, therefore, also of the number of obesity surgical interventions, their potential deleterious effects should be included in the differential diagnoses of unclear abdominal pain. PMID- 20020565 TI - [Andre Lenotre: master of illusion and splendor]. PMID- 20020566 TI - The passport system and state control over population flows in the Soviet Union, 1932-1940. PMID- 20020567 TI - Social disorder, mass repression, and the NKVD during the 1930s. PMID- 20020571 TI - Romiplostim: new drug. Thrombocytopenic purpura: for selected patients, with close monitoring. PMID- 20020570 TI - [Medical treatment and experiments in the Dachau concentration camp]. PMID- 20020572 TI - Lasofoxifene: new drug. Osteoporosis: no better than raloxifene. PMID- 20020574 TI - Ethical considerations about medical practices and the health team in relation to contributions from commercial firms. PMID- 20020573 TI - Overuse or misuse of thyroid function tests in pediatrics. PMID- 20020575 TI - Congenital hypothyroidism associated with neonatal tooth, Pierre-Robin syndrome and congenital heart defects. PMID- 20020577 TI - Association between serum levels of the soluble receptor (sRAGE) for advanced glycation endproducts (AGEs) and their receptor (RAGE) in peripheral blood mononuclear cells of children with type 1 diabetes mellitus. AB - AIM: The binding of AGEs to RAGE is involved in diabetic vascular complications. We studied sRAGE levels and RAGE protein expression (P) together with N carboxymethyl lysine (CML), a major AGE, in 74 patients with type 1 diabetes mellitus (DM1) and 43 healthy (C) children. METHODS: sRAGE and CML levels were determined by ELISA and RAGE P was evaluated in mononuclear cells by Western immunoblotting. RESULTS: Serum sRAGE was higher in DM1 than in C (1430 +/- 759 vs 1158 +/- 595 pg/ml, p = 0.047), inversely correlated to diabetes duration (r = 0.265, p = 0.037) and directly correlated to LDL-cholesterol levels (r = 0.224, p = 0.039). Diabetes duration correlated independently with sRAGE (p = 0.034). Circulating CML levels were not significantly different between DM1 and C groups (3.51 +/- 1.49 vs 3.59 +/- 1.83 ng/ml, p > 0.05) and RAGE P was lower in DM1 than in C (61 +/- 46 vs 102 +/- 63%, p = 0.0001). CONCLUSIONS: Increased serum sRAGE in children with DM1 may provide temporary protection against cell damage and may be sufficient to eliminate excessive circulating CML. PMID- 20020576 TI - A survey on Prader-Willi syndrome in the Italian population: prevalence of historical and clinical signs. AB - Clinical criteria for the diagnosis of Prader-Willi Syndrome (PWS) were established by consensus in 1993 (Holm et al.). Specific molecular testing is now available and the purpose of diagnostic criteria has shifted to identify individuals to test, thus avoiding the expense of unnecessary analysis. The aim of this study was to find clinical indicators to select patients with suspected PWS for laboratory testing. We analyzed the prevalence of clinical signs and symptoms in 147 genetically diagnosed Italian patients with PWS (67 males and 80 females), aged from 9 months to 34.6 years (13.6 +/- 8.3 years), using the consensus diagnostic criteria, and according to age, sex and type of genetic abnormality. The prevalence of several clinical features changed significantly with age, but very few with sex. According to genetic subtypes (deletion vs UPD), only hypopigmentation and acromicria were more frequent in patients with deletion. Some criteria considered as minor or supportive by Holm et al. have higher prevalence than some major criteria. In conclusion, in order to identify patients with suspected PWS to submit to laboratory testing, we recommend a classification of clinical criteria according to age, giving more attention to those so-called minor or supportive criteria. PMID- 20020578 TI - Adiposity is associated with endothelial activation in healthy 2-3 year-old children. AB - Adiposity is associated with C-reactive protein level in healthy 2-3 year-old children and with other markers of endothelial activation in adults, but data are lacking in very young children. Data from 491 healthy Hispanic children were analyzed. Mean age was 2.7 years (SD 0.5, range 2-3 years); mean body mass index (BMI) was 17.2 kg/m2 (SD 1.9) among boys and 17.1 kg/m2 (SD 2.1) among girls. E selectin level was associated with BMI (R = 0.11; p < 0.02), ponderal index (p < 0.02), waist circumference (p = 0.02), fasting insulin (p < 0.02), and insulin resistance (p < or = 0.05); these associations remained significant after adjustment for age, sex and fasting glucose. sVCAM was also associated with BMI (R = 0.12; p < 0.05). These observations indicate that adiposity is associated with inflammation and endothelial activation in very early childhood. PMID- 20020580 TI - Congenital hypopituitarism: clinico-radiological correlation. AB - BACKGROUND: Non-tumoral causes constitute a major group of childhood hypopituitarism. The structural abnormalities of the stalk and the pituitary gland correlate with number and severity of concurrent hormone deficiencies. We describe clinico-radiological correlates in patients with non-tumoral hypopituitarism. PATIENTS AND METHODS: Thirty-one children (23 boys) with congenital hypopituitarism, aged 1-17 years, with a peak GH levels of < 7 ng/ml after two pharmacological stimuli (insulin induced hypoglycemia [IIH] and clonidine) were included in the study. MRI tetrad of absent or hypoplastic pituitary, empty sella, redundant or absent stalk and absent/ectopic posterior pituitary bright spot (EPPBS) was considered as a radiological diagnostic criterion and these abnormalities were correlated with number of hormone deficiencies, severity of GH deficiency and mode of presentation at the time of delivery. RESULTS: Twenty (66%) children had vertex presentation, nine breech, and two children were delivered by lower segment Cesarean section (LSCS). Seven (78%) out of nine in the breech delivery group, 14 (70%) out of 20 in the vertex group and one out of two (50%) in the LSCS group had multiple pituitary hormone deficiencies (MPHD) (p = 0.665). Hypoplastic pituitary gland and posterior pituitary abnormalities were more frequent in patients with isolated growth hormone deficiency (IGHD) as opposed to MPHD (87.5% vs 65.2%, p = 0.08, 63% vs 47%, p = 0.64), whereas empty sella and stalk abnormalities were found more frequently in MPHD than in the IGHD group (76% vs 50%, p = 0.45 and 82.6% vs 37.5%, p = 0.01). Higher frequency of MR abnormalities was found in those with a peak GH response of < 3 ng/ml irrespective of the number of other pituitary hormone deficiencies (82.6% vs 37.5%, p = 0.02). Sixteen patients had MRI tetrad and it was more prevalent in the IGHD than in the MPHD group (75% and 44%, p = 0.01) and correlated with the severity of GH deficiency (r = 0.57, p = 0.01). The imaging abnormalities were also more prevalent in children with breech as compared to vertex presentation and correlated with severity of GH deficiency (100% vs 60%, p = 0.03, r = 0.52). CONCLUSION: Imaging abnormalities are frequent in patients with non-tumoral hypopituitarism and correlate best with severity of GH deficiency rather than number of hormone deficiencies and breech presentation. PMID- 20020579 TI - Homocysteine, fibrinogen and anti-ox-LDL antibody levels as markers of atherosclerosis in prepubertal obese children. AB - BACKGROUND: Homocysteine, fibrinogen and antibodies to oxidised LDL were shown to be important markers of atherosclerosis in adults. AIM: To investigate the levels of these three risk factors in prepubertal obese children. METHODS: Fasting homocysteine, fibrinogen and antibodies to oxidised LDL, plasma lipids, insulin, HbA1c and blood glucose levels were investigated in 30 prepubertal obese and 28 control children 6-9 years old. Investigations in the obese group were repeated after an oral glucose tolerance test. RESULTS: Fasting fibrinogen levels of the obese children were found to be significantly higher than those in the controls. Anti-ox-LDL antibody levels increased significantly after an oral glucose tolerance test. CONCLUSION: Fasting fibrinogen and postload ox-LDL levels which could act as important markers of coronary heart disease in later life could also be important risk factors in prepubertal obese children. PMID- 20020581 TI - The effect of 25-hydroxyvitamin D3 on the immune system. AB - AIM: To examine the effect of 25-OH vitamin D3 on the immune system in patients with nutritional rickets. METHODS: Fifty-three patients were included in our study between April 2002 and March 2004. Diagnosis of rickets was based on clinical, biochemical and radiological examinations. Cell surface markers (CD), complement factors (C), and immunoglobulin (Ig) levels were determined to find out any relationship between rickets and immune system deficiency. RESULTS: Among the causes of admission to hospital, fever (66%) and coughing (62.2%) were the most frequent. Pneumonia was accompanied by rickets in 47.1% of the cases. Plasma CD4 levels before the treatment were higher than those in the post-treatment period, whereas CD20 and CD56 levels were lower. CONCLUSION: B cell and natural killer cell reduction which occur because of vitamin D deficiency may contribute to the development of pneumonia and other infections in patients with nutritional rickets. PMID- 20020582 TI - Characterization of GH-1 mutations in children with isolated growth hormone deficiency in the Turkish population. AB - Isolated growth hormone deficiency (IGHD) is defined as a medical condition associated with growth failure due to insufficient production of growth hormone (GH) or lack of growth hormone action. It occurs with an incidence of between 1/4,000 and 1/10,000 live births. Most cases are sporadic and idiopathic but 5 30% of growth hormone deficiency (GHD) has genetic etiology. Mutations in the GH encoding gene (GH-1) have been detected in patients with IGHD. The purpose of this study was to characterize mutations of the GH-1 gene in children with IGHD in the Turkish population. We found four missense mutations (E33G, N47D, T-24A and A13S), one nonsense mutation (W-7X), one insertion and two deletions in nine patients out of seventy-five patients with IGHD. The missense mutation A13S, GAAA insertion at intron 1 (+178A), and the deletions of +83C in intron 1 and deltaF166 in exon 5 are novel mutations. PMID- 20020583 TI - Etiology and clinical features of isosexual precocious puberty in Taiwanese girls: twenty-three years' experience in National Taiwan University Hospital. AB - The aim of this study was to elucidate the etiology and clinical features of Taiwanese girls with isosexual precocious puberty. 460 girls with precocious puberty were enrolled in this study. 284 of them had a GnRH test and 179 girls with gonadotropin-dependent precocious puberty (GDPP) underwent brain MRI. Our results showed 7% of these 460 girls had gonadotropin-independent precocious puberty (GIPP). The other 93% patients had GDPP and 96% of them were idiopathic. Functional ovarian cyst and hypothalamic hamartoma were leading causes of GIPP and organic GDPP, respectively. The presence of undetectable basal serum FSH and LH levels had a specificity of 95% in girls with GIPP. Among girls with GDPP, those aged < 5 years had a higher percentage of detected CNS lesions than older girls (8% vs 3%). Girls with organic CNS lesions also had more advanced bone age SDS (6.9 +/- 3.3 vs 3.3 +/- 1.9), larger uterine volume (13.8 +/- 12.1 vs 6.6 +/- 8.1 cm3), higher basal serum estradiol (33.5 +/- 17.1 vs 26.8 +/- 18.2 pg/ml), and higher peak LH level after GnRH stimulation (45.2 +/- 36.2 vs 25.8 +/- 27.3 U/l) than those with idiopathic GDPP. In conclusion, GIPP is uncommon in Taiwanese girls with precocious puberty. Functional ovarian cyst and hypothalamic hamartoma are leading causes of GIPP and organic GDPP, respectively. The presence of neurological deficit, younger age at onset of puberty, presence of menstruation, rapid advance of bone age, markedly enlarged uterus, high serum estradiol, and high peak LH level after GnRH stimulation are suggestive of organic GDPP. PMID- 20020584 TI - Adiponectin gene polymorphisms in obese Greek youth. AB - We investigated single nucleotide polymorphisms 45T --> G and 276G --> T of the adiponectin gene in 48 obese Greek children and adolescents (3.58-16.25 years old) and examined their association with adiponectin levels and insulin resistance (IR), estimated with HOMA-IR, AUC(insulin) and WBISI. The polymorphisms were: 45T/G in 13/48 (27%) and 45G/G in 2 (4%) individuals; 276G/T in 21/41 (51%) and 276T/T in 3 (8%) individuals. Adiponectin in carriers of one or two G-alleles at position 45 was comparable to 45T/T (10.11 +/- 6.19 vs 8.03 +/- 4.96 microg/ml). Adiponectin in carriers of one or two T-alleles at position 276 was comparable to 276G/G (9.73 +/- 5.19 vs 7.77 +/- 5.65 microg/ml). SNP 45T -> G was not associated with IR. SNP276G --> T was associated with decreased risk for IR (OR = 3.05, 95% CI = 1.2384-11.13). In conclusion, SNPs 45T --> G and 276G --> T are common in obese young Greeks. SNP276G --> T might be protective against IR. PMID- 20020585 TI - Intractable neonatal seizures: an unusual presentation of congenital hypothyroidism. AB - Congenital hypothyroidism is the most common treatable cause of mental retardation. We report an unusual case of congenital hypothyroidism presenting as intractable seizures in an infant delivered to a mother known to have autoimmune hypothyroidism and who was noncompliant with therapy. To our knowledge, this rare presentation of congenital hypothyroidism has not been reported previously. PMID- 20020586 TI - Novel use of somatostatin receptor scintigraphy in localization of focal congenital hyperinsulinism: promising but fallible. AB - Congenital hyperinsulinism (CHI) is the most common cause of persistent hypoglycemia in infants and children. The pre-operative differentiation of focal from diffuse forms is extremely important from the management point of view. The current methods for pre-operative differentiation are invasive. We report a patient with CHI where somatostatin receptor scintigraphy was used to diagnose CHI but with limited success. A preoperative 68gallium DOTATOC-PET scan was performed which revealed highly localized radiotracer uptake in the body of the pancreas. 80% of the pancreas including the body was resected. The clinical problems did not completely resolve after surgery. Histopathology revealed hyperplastic islet cells at the resected margin and randomly throughout the pancreas. This case highlights the use of 68gallium DOTATOC-PET scan in a patient with severe CHI. The satellite foci that were missed may be either an inherent limitation of 68Ga DOTATOC scan or an underinterpretation due to lack of expertise. This report opens up a new option of using somatostatin analogue scintigraphy for pre-operative localization of hyperplastic islet cells in patients with CHI. PMID- 20020587 TI - 9 years follow-up of a patient with pituitary form of resistance to thyroid hormones (PRTH): comparison of two treatment periods of D-thyroxine and triiodothyroacetic acid (TRIAC). AB - Patients with pituitary resistance to thyroid hormones (PRTH) exhibit features of hyperthyroidism due to normal sensitivity to thyroid hormones in some of the peripheral tissues. There is a lack of information in the literature on the long term follow-up and treatment of patients with PRTH. Here, we present long-term (9 years) clinical and biochemical follow-up of a patient with PRTH under the treatment of D-T4 initially (for 1.5 years) followed by TRIAC for 5.5 years. An 11.5 year-old girl was evaluated for goiter, palpitations, heat intolerance, sleep disorders, nervousness and frequent stools for 3 years. Her thyroid function tests were consistent with PRTH. Molecular analysis revealed a heterozygous missense mutation of the TRbeta gene at codon 243 in exon 7 (R243Q) and a silent mutation at codon 245 in the index patient and the mother who was later also diagnosed to have PRTH. The patient was started on D-T4 treatment since she exhibited clinical symptoms of hyperthyroidism. After 1.5 years, D-T4 treatment was switched to TRIAC which lasted 5.5 years. During the long course of both treatments, thyroid hormones, TSH, heart rate, thyroid size, and markers of peripheral thyroid status (SHBG and alkaline phosphatase) were monitored. It was concluded that compared to D-T4, TRIAC treatment is more effective in suppressing TSH and lowering thyroid hormone levels in PRTH. However, both treatments were unable to reduce thyroid size. The effects of treatment on symptomatology were also modest. Spontaneous improvement in symptoms was observed with age. PMID- 20020588 TI - Short stature, celiac disease and growth hormone deficiency. AB - Celiac disease (CD) is a prevalent, genetically determined, autoimmune, chronic inflammatory state caused by intolerance to gluten that results mainly in gastrointestinal manifestations. One of the most common extra-intestinal manifestations of CD is short stature, and in some patients, short stature may be the presenting and only symptom of the disease, making the diagnosis of CD challenging. Impaired growth in children with CD results mainly from nutritional deficits, and withdrawal of gluten from the diet is frequently associated with a marked improvement of linear growth. In some patients, CD may be characterized by growth hormone (GH) resistance, as suggested by normal or elevated GH levels and low insulin-like growth factor-I (IGF-I) levels. Rarely, it has been shown that poor catch-up growth and/or IGF-I response to gluten-free diet may be due to the coexistence of celiac disease and GH deficiency. We present two children with coexisting CD and GH deficiency. One patient had MRI findings suggesting congenital isolated GH deficiency, and a possibility of developing multiple pituitary hormone deficiencies later in life. PMID- 20020589 TI - Clinical success of compomer and amalgam restorations in primary molars. Follow up in 36 months. AB - The purpose of this study was to evaluate the three-year clinical performance of a compomer and an amalgam in primary molars and to determine fluoride content in the enamel of these teeth after exfoliation. Twenty patients aged six to eight years randomly received one compomer and one amalgam restoration. Restorations were evaluated at baseline, 12, 24 and 36 months according to modified USPHS criteria. In six pairs of exfoliated teeth fluoride content at 1 and 5 mm distance from the restorations was determined. Replicas were made to determine marginal adaptation (MA) by scanning electron microscopy (SEM). At occlusal margins compomer restorations had better MA than amalgam restorations (p < 0.01). Compomer and amalgam restorations did not lead to different fluoride uptake in adjacent enamel and had an overall clinically satisfactory performance. The study confirmed that the assessed compomer is a suitable restorative material for class I and class II restorations in primary molar teeth. PMID- 20020590 TI - [Smokeless tobacco: a new risk factor for oral health? A review]. AB - Due to migration of different ethnic groups, mainly from Africa and Asia, into Switzerland, many traditional products and habits that have not been seen before were also introduced. Smokeless tobacco (ST)--as one of those habits--is a product of increased use in Switzerland, although no sound epidemiological data are presently available. Numerous studies from North-America, Sweden, Asia and Africa could verify the carcinogenic potency of smokeless tobacco and its effects in the developement of oral cancer and different systemic disorders. For dental professionals and their team it is important to detect ST-associated lesions, and to tell the patients about the potential harmful sequelae of ST products. The present review focusses on the different types of RT products and their effects on oral health. PMID- 20020592 TI - Culture and climate in health organisations. PMID- 20020593 TI - Understanding and improving patient safety: the psychological, social and cultural dimensions. AB - PURPOSE: This paper aims to provide researchers and practitioners with an overview of how organisation behaviour research (OBR) helps to understand and resolve patient safety problems in health care. DESIGN/METHODOLOGY/APPROACH: The paper describes psychological, sociological and other social science theories and research which help to understand the causes of patient safety problems, how to implement change effectively and how to create an organisational culture of safety. FINDINGS: Social science perspectives and organisational behaviour research are beginning to show why improvements in patient safety are slow, and how to make lasting and effective change. RESEARCH LIMITATIONS/IMPLICATIONS: Social sciences and OBR have already helped make healthcare safer, but could make a greater contribution. Progress could be faster with greater awareness of the findings of this research and understanding of social science research paradigms. PRACTICAL IMPLICATIONS: Better implementation and safer care could result from providing implementers and decision makers with more knowledge and access to social science research. More useful social science research could be developed by research funders and proposal reviewers gaining a greater understanding of social science methods and potential, and by including this research in a field made up largely of traditional experimental medical research methods. ORIGINALITY/VALUE: This paper provides an overview of the scientific and practical contributions of social sciences to patient safety and shows where future studies could assist understanding of current challenges and speed implementation of change. PMID- 20020591 TI - [Management of a failing implant supporting a bar-retained maxillary overdenture by means of alveolar ridge augmentation. Implant loss and reimplantation]. AB - This case report presents the treatment sequence of a 56 years old patient after he developed periimplantitis at the implant in position of tooth 22. This implant was integrated in an overdenture reconstruction connected to a soldered screw retained gold bar. The entire 2-stage procedure of implant explantation, simultaneous bone augmentation and new implant placement is documented. The onlay graft was performed by means of the Transfer-Ring-Control System (Meisinger). The existing gold bar could be resoldered and adapted to the new implant. Accordingly the overdenture was relined and the female retainer mounted. The treatment period covered almost one year. PMID- 20020594 TI - Analysing the "field" of patient safety employing Bourdieusian technologies. AB - PURPOSE: This paper aims to analyse the development of patient safety as a field within which patients are peripheral stakeholders. DESIGN/METHODOLOGY/APPROACH: The authors examined the patient safety movement from the perspective of a field in which agents struggle for control over various forms of capital, including economic, social, cultural and symbolic capital. In order to undertake this analysis the authors drew on the literature on errors and patient safety, key inquiries into patient safety, and research conducted with health professionals in New South Wales, Australia. FINDINGS: The patient safety movement has created a heightened sense of awareness of errors and risk across health systems, thereby attracting and creating significant amounts of capital. The authors argue that in the process of struggle to constitute and contain a new field of health, patients and their narratives are rendered vulnerable to appropriation and incorporation. RESEARCH LIMITATIONS/IMPLICATIONS: By considering patient safety from a sociological rather than a technical framework, it is possible to gain new insights into why reducing the levels of medical errors have proven so difficult. PRACTICAL IMPLICATIONS: Improved knowledge of how patient safety operates as a field may contribute to more effective strategies in reducing those types of errors. ORIGINALITY/VALUE: Despite the growth in the number of publications in patient safety there has been only minimal analysis of the field itself, rather than its technical or organisational components. This paper contributes to a new way of conceptualising and enacting patient safety, one that acknowledges the vulnerability of the parties involved, particularly patients. PMID- 20020595 TI - Positively deviant networks: what are they and why do we need them? AB - PURPOSE: This paper aims to report "positively deviant" experiences of three public sector networks seeking to enhance organizational and system level capacities. It is the authors' thesis that the knowledge base concerning the true benefits and pitfalls of networks can be captured and interpreted only through intense, ongoing learning effort embedded in practice on the ground, combined with sustained in-depth observation and collaborative research. DESIGN/METHODOLOGY/APPROACH: The paper describes through case examples why and how different kinds of networks within different jurisdictional contexts and different organizational cultures are being used to enhance the climate for change towards better health care and improved health. The authors describe the contexts, structures, processes and impacts of three "positively deviant" networks. FINDINGS: The network form can provide opportunity for nurturing changes and innovations within large organizational and complex system environments. This opportunity to create additional and different pathways for improved decision making and service provision comes with challenges that should be recognized. PRACTICAL IMPLICATIONS: The authors' experiences indicate that, for networks, a key component of success relates to pulling and pushing at the edges of multiple connections and boundaries in "positively deviant" ways. This pushing and pulling is intrinsically evidence of organizational and intraorganizational learning--in the examples presented--for the improvement of health care and health. ORIGINALITY/VALUE: Other networks can learn from the reported experiences and add their own cases to the empirical understanding of how networks can make a difference; this in turn can help the conceptual and theoretical understanding of them. PMID- 20020596 TI - Building knowledge integration systems for evidence-informed decisions. AB - PURPOSE: This paper aims to describe methods and models designed to build a comprehensive, integrative framework to guide the research to policy and practice cycle in health care. DESIGN/METHODOLOGY/APPROACH: Current models of science are summarised, identifying specific challenges they create for knowledge to action (KTA). Alternative models for KTA are outlined to illustrate how researchers and decision makers can work together to fit the KTA model to specific problems and contexts. The Canadian experience with the evolving paradigm shift is described, along with recent initiatives to develop platforms and tools that support the new thinking. Recent projects to develop and refine methods for embedded research are described. The paper concludes with a summary of lessons learned and recommendations that will move the KTA field towards an integrated science. FINDINGS: Conceptual models for KTA are advancing, benefiting from advances in team science, development of logic models that address the realities of complex adaptive systems, and new methods to more rapidly deliver knowledge syntheses more useful to decision and policy makers. PRACTICAL IMPLICATIONS: KTA is more likely when co-produced by researchers, practitioners, and policy makers. Closer collaboration requires shifts in thinking about the ways we work, capacity development, and greater learning from practice. ORIGINALITY/VALUE: More powerful ways of thinking about the complexities of knowledge to action are provided, along with examples of tools and priorities drawn from systems thinking. PMID- 20020597 TI - Organisational professional conflict and hybrid clinician managers: the effects of dual roles in Australian health care organizations. AB - PURPOSE: This article aims to examine tensions between hybrid clinician managers' professional values and health care organisations' management objectives. DESIGN/METHODOLOGY/APPROACH: Data are from interviews conducted with, and observation of, 14 managerial participants in a Cancer Therapy Unit set in a large teaching hospital in New South Wales, Australia, who participated in a Clinical Leadership Development Program. FINDINGS: The data indicate that there are tensions experienced by members of the health care organisation when a hybrid clinician manager appears to abandon the managerial role for the clinical role. The data also indicate that when a hybrid clinician manager takes on a managerial role other members of the health care organisation are required concomitantly to increase their clinical roles. RESEARCH LIMITATIONS/IMPLICATIONS: Although the research was represented by a small sample and was limited to one department of a health care organisation, it is possible that other members of health care organisations experience similar situations when they work with hybrid clinician managers. Other research supports the findings. Also, this paper reports on data that emerged from a research project that was evaluating a Clinical Leadership Development Program. The research was not specifically focused on organisational professional conflict in health care organisations. PRACTICAL IMPLICATIONS: This paper shows that the role of the hybrid clinician manager may not bring with it the organisational effectiveness that the role was perceived to have. Hybrid clinician managers abandoning their managerial role for their clinical role may mean that some managerial work is not done. Increasing the workload of other clinical members of the health care organisation may not be optimal for the health care organisation. ORIGINALITY/VALUE: Organisational professional conflict, as a result of hybridity and divergent managerial and clinical objectives, can cause conflict which affects other organisational members and this conflict may have implications for the efficiency of the health care organisation. The extension or duality of organisational professional conflict that causes interpersonal or group conflict in other members of the organisation, to the authors' knowledge, has not yet been researched. PMID- 20020598 TI - Critical pedagogy in a health service management development programme: can "critically thinking" managers change the NHS management culture? AB - PURPOSE: Management development programmes available to NHS managers focus on a performance orientation and sustain a culture of managerial and medical domination. This paper aims to question whether it is possible to consider NHS management development from a critical (empowerment culture) perspective. Features of the critical management studies approach (CMS) are identified. A new MSc is evaluated against these characteristics, examining the teaching and learning processes and students' perceptions of the programme. The aim is to develop critical thinkers who can return to their organizations and challenge existing power structures and practices to change local cultures and enhance health services. DESIGN/METHODOLOGY/APPROACH: Empirical research employed anonymous student questionnaires and a focus group. FINDINGS: Student evaluations suggest the MSc can deliver a critical pedagogy and help managers understand issues of power and empowerment, challenge dominant cultures, innovate and effect small, local changes in the NHS culture. RESEARCH LIMITATIONS/IMPLICATIONS: There is a need to continue evaluating the programme and include other stakeholders. Longitudinal research should assess the impact of the managers' changed values, attitudes and behaviours on colleagues, clients and the local cultures. PRACTICAL IMPLICATIONS: The paper identifies some of the tensions of developing "critical" health service managers, and the problems they encounter back in the "uncritical" NHS context, as well as some of the challenges in "facilitating" a critical curriculum. It questions the ethics of developing (or not) a critical perspective in a local context unfamiliar with CMS. ORIGINALITY/VALUE: Management development in the NHS largely ignores critical pedagogy. This paper makes a small and unique contribution to understanding how developing "critically thinking" managers can challenge the dominant culture. However, the limitations of such a small-scale study and ethical implications are noted. PMID- 20020599 TI - Socioeconomic indicators and risk of lung cancer in Central and Eastern Europe. AB - BACKGROUND: Social inequalities have been shown to contribute to the risk of lung cancer in industrialized countries, but it is unclear whether they also play a role in former socialist countries of Europe. METHODS: A case-control study involving 3,403 cases and 3,670 controls was conducted in Central European countries (Czech Republic, Hungary, Poland, Romania, Slovakia), Russia, and in the UK. Indicators of socioeconomic status, including education and white/blue collar occupation based on lifetime occupations were analysed as indicators of risk factors for lung cancer development, after adjustment for tobacco smoking and exposure to occupational carcinogens. RESULTS: Both indicators of socioeconomic status: low education and blue collar occupations were found as significant risk factors for lung cancer in men. The odds ratio of lung cancer for blue collar occupations compared to white collar occupations was 1.37 (95% confidence interval 1.15-1.62), that for low education compared to high education (analysis restricted to Central European countries) was 1.35 (95% confidence interval 1.03-1.77). No such effects were observed in women. CONCLUSIONS: The confirmation of the significant inverse association between the indicators of socioeconomic status and lung cancer risk in men may serve as a strong incentive for adoption of occupational and public health measures in lung cancer prevention. PMID- 20020600 TI - Factors influencing self-perception of health status. AB - The aim of this study was to evaluate subjective health status of Lodz adult population and to determine the factors affecting their self-perception of health. The study population consisted of randomly selected 1,056 adults aged 20 74 years from L6di district. Logistic regression model was applied to assess the factors influencing the self-perception of health. More than 30% of study subjects described their health as poor or very poor. There were no statistically significant differences between men and women regarding self-perception of health (p>0.05). Older people more frequently reported their health as poor and very poor compared to those younger than 25 years of age. Four percent of men and 10% of women younger than 25 years of age described their health as poor or very poor whereas in age category 45-54 years that percentage increased to more than 40% (men RR=16.3; p<0.001, women RR=7.5; p<0.001), in 55-64 to 60% (men RR=18.6; p<0.001, women RR=10.0; p<0.001) and for people older than 64 years of age to 60% for men (RR=12.6; p<0.01) and 72% for women (RR=13.4; p<0.001). People with lower educational degree perceived their health as worse compared to those with university diploma (men RR=5.3; p<0.001; women 4.6; p<0.001). The risk of indicating the health as poor or very poor was 3.4 times higher for unemployed men comparing to employed (p<0.001) and 1.5 for unemployed women compared to employed (p>0.05). Men indicating no leisure-time physical activity significantly more frequently described their health as poor or very poor than men with satisfactory level of recreational physical activity (RR=2.2; p<0.01). Current and former smoker men described their health as worse compared to non-smokers (current smokers RR=1.5; p>0.05; former smokers RR=1.8; p>0.05). Preventive programs aimed at improving self-perceived health should concentrate on increasing recreational physical activity and elimination of smoking. Those actions should in particular target people in older age category. PMID- 20020601 TI - Analysis of long term care in the context of social and health services in social institutional facilities in Slovakia. AB - OBJECTIVES: Long-term care for people with chronic illnesses and disabilities present an urgent challenge around the world. Existing systems of care, which typically rely on unpaid family members, are not by themselves enough to meet growing demands. This analysis includes survey on the structure of services provided in selected social facilities in Slovakia. METHODS: The survey on structure of provided services includes social facilities providing services to the target groups involved in this analysis: Elderly people in retirement age; Disabled and chronically ill citizens in working age. For data collection from social facilities we used questionnaire method. The data were obtained from the managements of these facilities (n=405). RESULTS: In social facilities, the age range of clients was 76-85 years (31%); the length of stay of a patient in a social facility was in 36% patients 11 years and longer. The most frequently performed activities in employees were related to services and activities (31%) and social care (30%). For 52% social facilities is waiting time 1 year and longer. The length of stay of patient in social facilities is affected in 83% by combination of health and social problems and as to the diagnosis structure the most prevalent conditions are ischemic heart diseases, heart attack, hypertension (39%), locomotor disease (36%). In 2006 the budget resources of social facilities were in 72% contributions from the state budget. CONCLUSION: Long-term care is closely interlinked with other programmes and systems, which can reduce the need for long-term care or which complement it. PMID- 20020602 TI - The "Five Risks algorithm": an easy tool for cardiovascular risk estimation. AB - The aim of this study is to provide an easy tool to identify patients with a high cardiovascular risk, especially those qualifying for lipid-lowering treatment. The decision to treat with lipid-lowering drugs was assessed with five new risk algorithms. The Five Risk algorithm (5R) takes into account male gender, high systolic blood pressure, high total cholesterol, smoking and high blood sugar as independent risk factors. Patients with three independent risk factors qualify for lipid-lowering treatment. Compared to the Framingham Risk Score, the 5R has a Kappa coefficient of 0.62. Compared to the SCORE, the Six Risk algorithm (6RDF) has a Kappa coefficient of 0.70. The 6RDF uses only four independent risk factors (male gender, high systolic blood pressure, high total cholesterol and smoking) but having diabetes or a family history of premature coronary heart disease are exclusion criteria for which treatment with lipid-lowering drugs is always indicated. PMID- 20020603 TI - Vertical distribution of the tick Ixodes ricinus and tick-borne pathogens in the northern Moravian mountains correlated with climate warming (Jeseniky Mts., Czech Republic). AB - A study of the vertical distribution of the common tick Ixodes ricinus and tick borne pathogens--tick-borne encephalitis virus (TBEV) and genospecies of Borrelia burgdorferi s.l.--was performed in the highest part of the Jeseniky mountain area (the Hruby Jesenik Mts. with the highest summit Praded, 1,491 m above see level). Altogether 1,253 specimens of all tick stages (607 larvae, 614 nymphs, 8 females and 24 males) were collected at the altitude 990-1,300 m above sea level on 12 collection sites by the flagging method. Altogether 1,207 ticks (8 females, 24 males, 568 nymphs and 607 larvae) were examined for the presence of tick-borne encephalitis virus and B. burgdorferi s.l. None of the samples contained TBEV, 35 samples (6% of adult ticks, 5% of nymphs, 0.7% of larvae) were positive for B. burgdorferi s.l. The most prevalent genospecies were B. afzelii (44%), B. garinii (28%), less frequent were B. burgdorferi sensu stricto (5%), B. valaisiana (3%). The rather large number of ticks (in absolute numbers as well as recounted to the index: average number of nymphs/worker/collection hour) and the presence of all developmental stages clearly demonstrate that there are viable local tick populations in all the sites, and that recorded ticks were not randomly individuals brought into higher altitudes by birds or game animals. The results are compared with the long-term (2002-2007) monitoring of the tick altitudinal distribution in the Krkonose Mts. and the conditions, which allow ticks to establish local populations up to the timberline in both mountain areas, are discussed. Simultaneously, changes in climatic conditions (especially the air temperature) monitored at 3 meteorological stations in the area of the Jeseniky Mts. were compared with the records from another 8 stations in other mountain areas in the Czech Republic. A very similar statistically significant trend of increasing mean air temperatures during the last three decades is found at all analyzed stations. The trend is most pronounced in the spring and summer months with the highest activity of I. ricinus ticks. PMID- 20020604 TI - The reform of specialist training for general practice in the Czech Republic. AB - AIMS: The aim of this paper is to describe the reform of General Practitioners' (GP) training in the Czech Republic (CR) and compare it with the recommendations of European institutions of General Practitioners and European Union (EU) legislation. METHODS: The structures of the new Czech GP training (2007) were compared with the recommendations of The European Union of General Practitioners (UEMO), European Academy of Teachers in General Practice and the requirements set by the EU legislation. The comparison with other countries was based on data from the UEMO questionnaire distributed to its members. Data from representatives of institutions responsible for specialist training in the CR and from statistical offices in the CR were also used. RESULTS: The GP specialist training in the Czech Republic does not differ significantly when compared to the other UEMO countries, however, not all the recommendations of the European GP associations have been fulfilled. CONCLUSION: The reform of GP specialist training brought about positive changes but further developments are still needed. In our opinion, the next necessary step is the setting up of quality criteria for the assessment of GP specialist training and of the primary care as a whole. PMID- 20020605 TI - Arsenic contamination in environment in the region of Vojvodina. AB - Ground waters in the region of Autonomous Province of Vojvodina, Republic of Serbia are endangered by arsenic (up to 0.750 mg/l). Total arsenic concentration was determined In samples of untreated and treated water from some local and central water-supply systems. Results are compared to actual regulations in the country. This analysis encompassed 324 drinking water samples of various sources, analyzed in the Institute of Public Health Novi Sad, during 2005. Determined concentration of total arsenic in drinking water varies from 0.005 to 0.450 mg/l. Arsenic concentration in the river Danube water was within recommended value for I-II class. Maximum arsenic daily intake through food and nutrition was 60.9 +/- 22.3 microg/day in 2000. To understand importance of the problem of arsenic environment contamination in Vojvodina region, our own results, as well as the results of other authors are presented and analyzed (drinking water: 173 samples, moss deposition and daily intake through nutrition). PMID- 20020606 TI - A prospective study of acute poisonings in a sample of Greek patients. AB - OBJECTIVES: Acute poisoning is one of the most common situations managed in the emergency department. Significant differences have been reported concerning the prevalence and etiology of poisoning. This study aims to present the epidemiologic and clinical characteristics of poisonings in the region of Thrace in Northern Greece, where no similar studies have been performed. METHODS: A prospective study was carried out in our hospital's emergency department during 4 years (1999-2003). Atotal of 223 adult patients were admitted with acute poisoning, 87 males (39%) and 136 females (61%). RESULTS: Mean age of male and female patients was 37.1 years (standard deviation 16.1) and 33.4 years (standard deviation 14.5), respectively. Intoxications were more common in summer (34.1%). The median time between poisoning and presentation was 4 hours. Poisonings were more common in the urban population (64.1%). Psychotropes were the leading cause of poisoning in patients with a psychiatric disease (74.1%) and analgesics in all the other patients groups (34.8%). The clinical status on admission was: conscious (45.7%), confused (35.4%), precoma/coma (18.9%). Haemodialysis was performed only in one patient and respiratory support was necessary in 10 patients. An antidote was administrated in 73 patients. Median hospital stay was 1 day. Two patients died (0.9%). CONCLUSIONS: Poisoning's aetiology varies significantly in different countries. Previous suicide attempts are common in these patients. Precipitating factors can be major depressive, or dysthymic disorders. The mortality rate is low. PMID- 20020607 TI - The impact of teaching physical education to Czech school children using progressive teaching approaches: findings of a four-year study. AB - Practice teaching is an important feature of pre-professional preparation of teachers. This study assessed student teachers' (STs) and pupils': overall evaluation and evaluation of pupils' roles in physical education (PE) and second subject lessons. STs delivered the lessons using either traditional or progressive teaching approaches. Questionnaires were completed by 57 STs and 10,517 high school pupils who assessed the lessons taught by the STs. Differences in the evaluation of pupil's role were analysed according to teaching approach, school years, and gender. In both types of lessons, pupils evaluated progressive teaching approaches more positively than the traditional approaches. Pupils' roles in lessons delivered using progressive teaching approaches also increased. STs evaluated the progressive approaches in both lessons more positively than traditional approaches. Girls evaluated PE lessons more favourably than boys regardless of teaching approach. If the goal is to increase pupils' role in the lessons, progressive teaching approach may be more effective than traditional. PMID- 20020608 TI - More people dying from TB are HIV-infected than previously recognized. PMID- 20020609 TI - [Conceptual approach to formation of a modern system of medical provision]. AB - Within the frame of forming of a new face of medical service of the Armed Forces, were determined the principle approaches to optimization of the process of development of the system of medical supply. It was proposed to use the following principles: principle of hierarchic structuring, principle of purposeful orientation, principle of vertical task sharing, principle of horizontal task sharing, principle of complex simulation, principle of permanent perfection. The main direction of optimization of structure and composition of system of medical supply of the Armed Forces are: forming of modern institutes of medical supply- centers of support by technique and facilities on the base of central, regional storehouses, and attachment of several functions of organs of military government to them; creation of medical supply office on the base military hospitals, being basing treatment-prophylaxis institutes, in adjusted territorial zones of responsibility for the purpose of realization of complex of tasks of supplying the units and institutes, attached to them on medical support, by medical equipment. Building of medical support system is realized on three levels: Center - Military region (NAVY region) - territorial zone of responsibility. PMID- 20020610 TI - [Organization of scientific-methodological work in Central Military Clinical Hospital named after A.A. Vishnevskii]. AB - There was congested a wide experience of organization and management of scientific work in the 3rd CMCH by Vishnevsky A.A. for a term of more than 40 years. This experience is subjected to generalization, analyze for the purpose of determination of it's priority orientations of improvement. Scientific-methods work in hospital is rated as a complex of measures, organisationaly-planed and coordinated by purpose and reinforcement of scientific schools of the 3rd CMCH by Vishnevsky A.A., as a basis of effective delivery of specialized medical aid. The vector of scientific researches is directed, generally, to solving questions of military and field medicine. PMID- 20020611 TI - [Incendiary and toxic characteristics of phosphorus. Lessons learned from the accident in L'vov]. AB - The article presents the analyze of cause of the accident of freight train by the 16th of July 2007 on Lvov railway, when 15 cisterns with yellow phosphorus have went off the rails. At the moment of the accident there was broken gasproofing of 6 cisterns, from which liquid phosphorus has started to supervene. In firefighting have taken part 450 firemen, 3 firetrains, 80 units of other techniques. The fire was put out for 5 hours. 58 military servicemen and 6 journalists have got hurt. Besides fire hazard, it was formed a nidus of infection for 80-90 km2. 2484 persons were exposed to phosphorus fume. 184 from them were sent to hospital, 800 were evacuated, 1500 children were sent to sanatoriums. In the article were observed clinical forms of phosphorus affect, organizational aspects of delivery of health care in conditions of mass destruction in fire hazard of phosphorus. PMID- 20020612 TI - [Effect of imunofan on several indexes of immunity and pyoderma incidence among cadets of educational institutes]. AB - It was discovered that during the first several months military service men have suppression of several indexes of immunity and a high level of morbidity by pyoderma. Was shown a high effectiveness of immunomodulatory agent Imunofan on correction of immunity status of servicemen and decrease of morbidity by pyoderma (more than on threefold): the effectiveness index is 3,25, the defense index is 69,2%. In addition, it was marked hepatitis- and stress-protect effect of the agent. It was made a conclusion about necessity of further examination of questions of correction of immunity status of military servicemen for the purpose of decrease of morbidity by infections, actual for studying units of the Armed Forces of RF. PMID- 20020613 TI - [Damages of ENT-organs inflicted by the weapon of not-lethal (traumatic) action]. AB - There was effectuated a research of killability of traumatic weapon in laboratory as well as during analyzing of clinical observations. There were considered the results of treatment of 8 persons, suffered from traumatic weapon. It was occurred a clinical recovery by 7 wounded persons from 8. There was 1 case of lethal outcome for first hours, caused by trauma of temporal bone and cerebrum (suicide). Thus, influence of traumatic weapon on ENT-organs can be a reason of serious penetrating wounds, facial bones fracture, hematomas and contusion, formation of foreign bodies and middle ear barotraumas. PMID- 20020614 TI - [The 20th anniversary of Urological Center of Main Military Clinical Hospital named after N. N. Burdenko]. AB - In 1988, for the first time according to Directive of Central Command of Armed Forces of USSR in Main Military Clinical Hospital by Burdenko N.N. was founded a interhospital unit - Urological center. The article presents the analyze of work of the Center during 20 years. The most frequent diseases--adenoma of prostate, malignant swelling and benign tumour of urological organs, urolithiasis, traumatic damages of urological organs. During a year 4500-4700 persons are treated, 1450-1500 are operated. Every year 44-77 military medical men are studying, 20-26 clinical attending physicians study for 2 or 3 years. During last 8 years 422 military medical men and 68 clinical attending physicians have studied. PMID- 20020615 TI - [Interdependence between alcohol consumption and mortality from hepatic cirrhosis]. AB - The article presents the data on the dependence between death rate caused by cirrhosises and consumption level of alcohol in Europe and Russia, based mainly on data of foreign publications, particularly--the results of ECAS research with a mathematical description of the taped rigid dependence between these indexes. The article also presents short characteristic of alcohol situation and executability of equations of regress, received from ECAS, for levels of alcoholic consumption and death rate from liver cirrhosis in Russia. PMID- 20020617 TI - [Evaluation of functional adaptation level in air specialists according to biochemical indexes of saliva secretion]. AB - It was examined a capability of evaluation of functional condition of air staff by indexes of natrium, kalium, cortisol and glucose in saliva. There were realized 5 series of examinations with participations of 71 airplane pilot of the same level in conditions of realizing flies of different difficultness. Saliva sampling was effectuated before and after the flies not later then 10-15 minutes after landing. On pre-flight medical examination and after performance of task of air relay there was registration of systolic, diasystolic blood pressure and cardiac rate. It was posed the correlation of physiological indexes with percentage of examined ingredients in saliva in different flight loads. The results of examinations speak for capability of using of indexes of percentage of natrium, kalium, cortisol and glucose in saliva for evaluation of functional condition of airplane pilots during effectuating the flies and rating of value of flight load with account of individual peculiarities. PMID- 20020616 TI - [Organizational-epidemiological aspects of prophylaxis of outhospital pneumonia in military communities]. AB - The article presents actual questions of epidemiology of outhospital pneumonia (OP), peculiarities of epidemic process of these infections in troops in now-day conditions and updates of main directions of their prophylaxis. It was marked that alongside with clearly defined tendency of lowing, the morbidity by OP in the Armed Forces of RF is still rather high--in 5-7 folds higher than among adult population of the country. Whereby it is predicted further complication of sanitarium-epidemiological situation in connection with increase of call-up. The article uncovers the main factors, supplying the realization of mechanism of epidemic process of OP in military collectives. Vaccination of staff by registered in RF multipartial pneumococcal vaccine Pnevmo-23 is considered as an effective and necessary medical measure in complex of prophylactic measures, effectuating in troops. PMID- 20020618 TI - [Cholera and war]. AB - During last centures wars were the main account of spread of cholera. It is caused by movement of great mass of troops and peaceful populace, acute fall of living circumstances, decline of sanitarium conditions of population aggregates, difficultness or impossibility of effectuating of contra-epidemic measures. Cholera casualty was multifold bigger, the weapon casualty in fighting armies. The article presents data of cholera epidemics, were in fighting armies of different states. During the XXth century fight casualty began to overpass the disease casualty. It is caused by grand increasing of damage effects of measures of war, organized using of prophylaxis measures and success in treatment of infectious diseases. The article presents data about cholera falling ill during the Great Patriotic War and about system of contro-epidemic barrier on fronts and rear of state. PMID- 20020619 TI - [The 75th anniversary of Military Sanatorium "Volga"]. AB - The article presents the history of development and becoming of a military sanatorium Volga. In the article was shown the structure of sanatorium and units of rehabilitation treatment, was shortly observed an individual program of rehabilitation of military servicemen. There were marked veterans of sanatorium, who have been working more than 30 years, and family dynasties, which works now days. PMID- 20020620 TI - [Analysis of direct results of mitral valve annuloplasty]. AB - In the present-day cardiosurgery there is no doubt that in case of possible plastic operation on the mitral valve the latter must be preserved. In the current literature there are no investigations which could convincingly prove advantages of any single group of techniques of constriction of the fibrous ring when performing mitral reconstructions. The aim of the work was to compare the direct results of reconstructive operations on the mitral valve with the using of three different means of constriction of the fibrous ring. It was found that all the three types of plasty of the mitral valve fibrous ring gave good direct results. Nevertheless an analysis of long-term results is necessary for the assessment of effectiveness of mitral reconstructions in the long-term postoperative period. PMID- 20020621 TI - [The role of tumor markers in diagnostics of gastric cancer metastases. Part II: postoperative monitoring]. AB - Complex examination of 32 patients with gastric cancer in terms of more than 6 months after radical operations included endoscopic, radial and laboratory methods with the determination of indices of the carcinoembryonic antigen and carbohydrate antigens 19-1 and 72-4 every six months. It was established that normal concentration of all the three tumor markers within 12 months after surgery was a practically reliable sign of the absence of metastases and recurrence of gastric cancer. The increased level of one or two oncomarkers within 12 months after radical operation points to progression of gastric cancer with probability of 57.1% or the development of other surgical pathology (cholecystolithiasis, gastric stump polyps and others) in 28.6% of observations. In all cases of progressing gastric cancer in terms of more than 6 months after operation there was a more that 1.5 times elevation of any one or two tumor markers. PMID- 20020622 TI - [The prognostic role of interleukin-6 in peritoneal exudate of patients with diffuse peritonitis]. AB - A prospective longitudinal controlled investigation of 189 patients with secondary diffuse peritonitis was performed. A correlation between the levelof interleuikin-6 of peritoneal exudate, the severity of the patients' state, the number of complications and outcome of the course of the disease was established. The maximal number of complications and unfavorable outcomes falls on the peak value ofinterleuikin-6. PMID- 20020623 TI - [Prophylaxis of acute pancreatitis during resection of the stomach for "difficult" complicated of duodenal ulcers]. AB - The authors have performed 407 resections of the stomach for duodenal ulcers. In 9 cases the major duodenal papilla was found to be involved into the zone of ulcer destruction. It was noted that operations for "difficult" complicated ulcers of the duodenum with the involvement into the zone of destruction of the major duodenal papilla required a delicate surgical strategy directed to minimization of trauma and blockade of the main factors of the development of acute postoperative pancreatitis, the proximal duodenojejunoanastomosis being the operation of choice for "difficult" complicated ulcers of the duodenum. PMID- 20020624 TI - [Causes of recurrences of gastrointestinal ulcer bleeding, their prophylaxis and treatment]. AB - The authors have analyzed main causes of recurrences of gastro-intestinal ulcer bleeding which are thought to be the continuing ulcer alteration, lysis of the protective thrombus with gastric juice or the fibrinolysis system. The data obtained allowed determination of laboratory signs of the development of DIC syndrome and its significance in the appearance of recurrent bleeding and greater lethality. The role of antirecidivation antisecretory therapy and therapeutic endoscopy was studied for prevention of recurrent bleeding depending on the degree of blood loss. The indications were formulated for emergency, postponed and planned surgery for gastro-intestinal ulcer bleedings as well as the content of conservative antirecidivation therapy. PMID- 20020626 TI - [Combined surgical interventions for advanced thyroid carcinoma]. AB - An analysis of specificities of 141 combined operations for advanced (T4) papillary, follicular and medullary thyroid carcinoma has shown that such operations proved to be necessary in 5.4% out of 2606 patients operated for carcinoma of the thyroid gland (CTG). In most cases these operations were of organ-sparing character. In 10.5% of the patients circular resections of organs of the respiratory and digestive systems were fulfilled on the neck and head. Complete removal of the tumor was obtained in more that 2/3 of observations. The direct and long-term results were noted after thyroidectomies, lymphadenectomies and organ-sparing resections of other neck organs. The indices of 5 and 10 years survival of patients with papillary, follicular and medullary thyroid carcinoma depended on gender, on the tumor affected organ, on the histological type of the neoplasm under conditions of performing the operations by highly professional surgeons. PMID- 20020625 TI - [Surgeon's strategy in forming large intestine anastomoses]. AB - The article includes experience with treatment of 103 patients with the formed different large intestine anastomoses. Primary operations for cancer of the rectum were made on 76 patients, restorative operations--on 27 patients. The following techniques were used: manual formation of the large intestine anastomosis, apparatus anastomoses using AKA-2, "ETHICON CDH" and double apparatuses method using "CONTOUR" and "ETHICON CDH". It was found that the application of stitching apparatuses required shorter time necessary for applying large intestine anastomosis and for operation. When forming the large intestine anastomoses in the abdominal cavity the manual method should be preferred. The formation of anastomosis in the small pelvis cavity is accompanied by technical problems and requires using stitching apparatuses. The method using apparatuses "CONTOUR" and "ETHICON CDH" decreases the number of postoperative complications and can extend the list of indications for performing sphincter-sparing operations. PMID- 20020627 TI - [Central lymphodissection in carcinoma of the thyroid gland]. AB - Experience with central lymphodissection (CLD) was analyzed in 93 patients with primary thyroid cancer (pT1-4aN0-1a). Prophylactic central lymphodissection was fulfilled in 48 patients, therapeutic CLD in 45 patients. The size and spread of the primary thyroid tumor had no difference in both groups of patients. In patients of the 2nd group thyroidectomy was fulfilled more often (71.1%) than in the 1st group (58.3%).The frequency of postoperative transitory hypoparathyroidism after surgery on the thyroid gland and therapeutic CLD made up 13.3%. There were no recurrent metastases of cancer after CLD fulfilled during the last 3.5 years. All cancer metastases in the postoperative period were detected in lymph nodes of the neck (2-5 levels) and their prevalence was 8.9%. PMID- 20020628 TI - [Treatment of disrupted pelvic ring in fertile women]. AB - The article presents the data on frequency of disrupted pelvic ring in fertile women on the basis of the Djanelidze Research Institute of Emergency Medicine database from 2000 to 2005. Strategy of treatment, target dates of the pelvis fixation and removal of fixators are described depending on planning the pregnancy terms. In addition, approaches to childbirth, indications and contraindications to Caesarian section were studied on the basis of the Snegiryov maternity hospital database for the period from 2000 to 2006. PMID- 20020629 TI - [Use of a diode laser in surgery of intervertebral disks in osteochondrosis of the vertebral column]. AB - The results of surgical treatment of 185 patients with osteochondrosis of the vertebral column were studied during 12 months who, after complex clinico-radial examination, were treated by puncture laser nucleoplasty of intervertebral disks using a diode laser. Stable positive effect was obtained in 84.3% of cases using minimally invasive manipulations. PMID- 20020630 TI - [Intraoperative monitoring and its practical value in operations for tumors of the posterior cranial fossa in children]. AB - The work is based on an analysis of the course of anesthetic aid, operative and early postoperative period in 62 patients aged from 15 months to 18 years with tumors of the posterior cranial fossa. The analysis concerned changes to the indices of systemic hemodynamics, findings of electrophysiological monitoring (electroencephalogram, acoustic stem induced potentials) and their interrelation with the outcomes of surgery. It was found that permanent disturbance of the heart rate such as bradycardia and tachycardia as well as the decreased indices of latency of peaks of the acoustic stem induced potentials and inter-peak intervals registered at the stage of removal of tumors of the posterior cranial fossa must be taken into consideration in the estimation of indications to medicinal narcosis in the postoperative period. PMID- 20020631 TI - [Development of the method of high frequency insufflation of medical aerosol for treatment and prophylaxis of the adhesion process in the abdominal cavity]. AB - A method of treatment of the abdominal cavity in peritonitis patients has been developed which promotes early regeneration of the peritoneum mesothelium, fibrinolysis, recovery of functions of the abdominal cavity and thus preventing formation of the adhesion process in the abdominal cavity. Effectiveness of the method was experimentally confirmed in rabbits and used in 126 patients with different forms of peritonitis. A comparison of methods of traditional treatment of peritonitis in 120 patients and that with using the developed method has shown advantages of the latter one. PMID- 20020632 TI - [Experience with using anterior resections in the surgery of rectum]. AB - A retrospective analysis of direct results of anterior resections of the rectum fulfilled for cancer is presented. The patients were divided into 3 groups according to the methods of forming colorectal anastomoses: manual nodular, compression-mechanical method using apparatus AKA-2, 4 and a compressive method with apparatuses on the basis of the effect of memory of the form in the titanium nikelide alloy. Advantages of using compressive apparatuses are shown as well as the reliability of compressive colorectal anastomoses. PMID- 20020633 TI - [Minimally invasive surgery of pancreonecrosis--progress and problems]. AB - An analysis of treatment of 133 patients with acute pancreatitis using minimally invasive surgical interventions (laparoscopic and puncture-draining operations) has shown that in cases of total pancreonecrosis these methods were little effective in most cases as a definitive method of surgical treatment and may be considered as the first stage of complex process of treatment. However, for certain forms of pancreonecrosis the minimally invasive surgery plays one of the leading roles in the improvement of results of treatment in the general population of patients with destructive pancreatitis. PMID- 20020634 TI - [Application of the bifurcation stent-graft ELLA in endovascular treatment of the abdominal aorta aneurysms]. PMID- 20020635 TI - [Combined treatment of primary multiple metachromatic cancer of the lung and large intestine]. PMID- 20020636 TI - [Intestinal invaginations in patients of the older age group]. PMID- 20020637 TI - [A rare abnormal development of the thoracic part of the esophagus in an adult]. PMID- 20020638 TI - [Postsplenectomy sepsis]. PMID- 20020639 TI - [The Ogilvie syndrome after restorative surgery on the large intestine]. PMID- 20020640 TI - [From "Glacial anatomy" to computed tomography]. PMID- 20020641 TI - [Professor Mikhail Vasil'evich Alferov (1883-1941)]. PMID- 20020642 TI - [Academician Mir-Asadulla Mir-Alekser ogly Mir-Kasimov (1883-1958)]. PMID- 20020644 TI - [The diagnosis and strategy of complicated esophagitis treatment]. PMID- 20020643 TI - [Certain possible complications and side effects after the introduction of interleukin-2 in the scheme of treatment of surgical patients]. PMID- 20020645 TI - [Cell cycle alterations and senescence in response to DNA damage]. PMID- 20020646 TI - [Circulating stem cells: a comparison of breast cancer and cardiovascular pathology]. PMID- 20020647 TI - [Standardized indices of cancer morbidity based on data from national cancer registries: significance and causal relations]. PMID- 20020648 TI - [Oncologic aid to colon cancer patients (population study)]. PMID- 20020649 TI - [Oncological service in the Republic of Belarus in 2008 and prospects for development]. PMID- 20020650 TI - [Can extended induction chemoradiotherapy be effective in the combined treatment of stage IIIB Hodgkin's disease ?]. AB - The paper evaluates the results of chemoradiaton (2-4 cycles of preliminary treatment to suppress intoxication symptoms plus total, or subtotal exposure of the lymph nodes to tumor-killing doses in cases of intact iliac inguinal nodes) in 89 patients with Hodgkin's disease stage IIIB. The data are evaluated vis-a vis regression in exposed foci as intoxication symptoms receded. Overall survival was significantly lower (p = 0.044) when tumor regression was well below 75%. Similar trends were reported with regard to relapse-free survival. It is concluded that, with our regimen, combined treatment will not be effective unless 75% or more regression of tumor is assured. PMID- 20020651 TI - [Improved high-frequency flow ventilation-assisted endotracheobronchial surgery for tumors of the trachea and main bronchi]. AB - The report deals with evaluation of the literature and our own data on 65 endotracheobronchial operations for tumor-related obstruction of the main bronchi using high-frequency injection flow ventilation (n = 33; 50.8%) or a combined frequency one (n = 32; 49.2%). The latter method was significantly more efficient (p < or = 0.05%) due to more effective blood oxygenization (pO2--111.8 +/-13.8 vs. 130.6 +/-26.4 mmHg) and carbon dioxide elimination (pO2--36.1 +/-1.2 vs. 54.3 +/-6.7 mmHg). It also involved lower risk of surgical fire as oxygen concentration in inhaled air mix was monitored. Also, endoscopic examination took less time because it was continuous, while the risk of barotrauma was reduced to minimum owing to adequate selection of ventilation ratings. PMID- 20020652 TI - [Use of optical coherence tomography for prognosis of the severity of oral mucositis]. AB - The results of oral mucosa monitoring by optical coherence tomography (OCT) in the course of radiochemotherapy of 18 cases of oropharyngeal cancer are discussed. Damage to the mucosa was mainly assessed using contrast characteristics deterioration and reduction of epithelial layer thickness. Significant variation in OCT image characteristics was identified vs. mucositis grade and prognostic criteria for individual mucosal radiosensitivity worked out intact contrast on day 1 of mucositis should be interpreted as a sign of relatively mild complication. Blurred contrast would indicate mucositis stage III IV development. Numerical analysis of OCT image patterns confirmed the contribution of the endothelial blood vessels and connecting tissue to radiation induced damage to the mucosa. PMID- 20020653 TI - [Influence of immunological parameters on the effectiveness of systemic and loco regional immunotherapy in disseminated renal carcinoma]. AB - Survival was assessed among patients with disseminated renal carcinoma from different prognostic groups as established by MSKCC protocol. Multivariate evaluation pointed to prognostic value of IL-6 (spontaneous and induced production), IL-8 (spontaneous and induced production),TNF-alpha (spontaneous production), IFN-gamma (induced production), TNK-cells (CD3+CD16+CD56+), and T regulatory cells (CD4+CD25+Treg). It was demonstrated that additional criteria may be used to evaluate prognosis. PMID- 20020654 TI - [Sclerosing epithelial fibrosarcoma: a clinico-morphological evaluation of 10 cases]. AB - Sclerosing epithelial fibrosarcoma is a malignant mesenchymal tumor (fibroblastic class). It involved tissues of the legs and trunk in patients aged 31-59 and grew slowly as an infiltrative neoplasm during 6-18 months. Relapse occurred in 20% although no metastases were reported. Tumor consisted of epithelial fibroblasts with light cytoplasm and mononuclei. The latter showed low mitotic activity. Fibroblasts came in clusters, bands and alveolar formations sitting in the fibrous stroma. There were small areas consisting of bundles of spindle-shaped fibroblasts with oval hyperchromatic nuclei. Differentiated analysis of epithelioid fibrosarcoma may also be done by computed microscopy and immunomorphology using other tumor models consisting of light-cytoplasm epitheloid cells, mostly osteosarcoma, clear-cell renal carcinoma, melanoma, synovial sarcoma and clear-cell sarcoma of the tendon sheath and aponeurosis. PMID- 20020655 TI - [Radiologic diagnosis as a component of staging and surgical approach in carcinoma of the renal parenchyma]. AB - Due to its long-term symptom-free course, carcinoma of renal parenchyma would be detected by chance in 50% of cases. Clinically, advanced tumor was diagnosed in 85%. The report deals with criteria of screening for surgical treatment used in 215 cases following preoperative examination. PMID- 20020656 TI - [Kinetic features of ascites and solid Ehrlich tumors]. AB - Such biological parameters as tumor volume, Ki-67 and p53, which characterize the development of ascites and solid tumor of Ehrlich were evaluated. The kinetic curve of growth of ascites tumor was S-shaped (Gomperts) while that of the solid one--cubic (Speer-Retsky). Ki-67 expression level was found to be in cyclic correlation with duration (3 and 6 day intervals) which might be worth considering when working out therapeutic procedure. Moreover, no increase in cell death was observed when tumor growth slowed down. PMID- 20020657 TI - [Effect of sutent and celecoxib on the properties of endothelial cells in vitro]. AB - We studied the anti-angiogenic properties of sutent (SU11248) and celecoxib in human endothelial cell line EA. hy 926 in vitro. Sutent 0.05-0.5 microg/ml suppressed their proliferation and migration depending on dose while celecoxib did the same at 5.0 microg/ml. We were the first to demonstrate that endothelial cell incubation was followed by increase in 5'-nucleotidase activity in the presence of sutent while celecoxib did not produce such effect. It may be suggested that elevated 5'-nucleotidase concentration at the membranes of endothelial cells might in turn contribute to the pool of extracellular adenosine to stimulate antiinflammatory effect. Our data also contribute to the knowledge about the anti-angiogenic properties of sutent and celecoxib. PMID- 20020658 TI - [Diethylnitrosamine-induced carcinogenesis in PARP-1(-/-) and PARP-1(+/+) mice]. AB - Our study is concerned with comparative analysis of diethylnitrosamine (DENA) induced carcinogenesis in PARP-1 knock-out female mice PARP-1(-/-) and wild type animals PARP-1(+/+). No difference was recorded in relation to total tumor incidence (88 and 95%, respectively): cardia (87 and 84%, respectively), liver (80 and 66%, respectively). However, experimental animals PARP-1(-/-) tended to reveal incidence of cardia tumors with invasion as deep as the serosa higher than in PARP-1(+/+) mice (100 and 81%, respectively) and metastases to the liver and lung--27 and 7%, respectively. Relative incidence of angiosarcoma and holangiocarcinoma among liver tumors from PARP-1(-/-) mice was higher than that in wild type mice. Hence DENA induced the most aggressive tumors in PARP-1(-/-) knockout mice more often than in PARP-1(+/+) ones. Our results confirm the significance of the role of DNA repair in carcinogenesis. PMID- 20020659 TI - [Transfer factor specific to mouse lung carcinoma of Lewis]. AB - Factor of transfer was identified and sampled in the course of immunization of rats with cells or tissues of mouse lung carcinoma of Lewis and its antitumor action investigated on models of passive and spontaneous dissemination in mice C57BL/6. Samples obtained at the peak of immunological response (day 14) and immunological memory inception (day 60) were shown to be capable of marked antimetastatic effects: fall in metastatic frequency, number and size of metastases correlated with higher index of metastasis formation. PMID- 20020660 TI - [Treatment of hepatic malignancies with implantation of a venous port into the portal vein]. AB - The report deals with venous port implantion into the portal vein to conduct both single and continuous courses of regional chemotherapy without recourse to repeat puncture and catheterization of the vein to avoid hemorrhagic complications and arterioportal fistula. Application of repeat courses of intraportal chemotherapy might make use of ultrasound and X-ray equipment to monitor the catheter position in portal vein lumen as well as unauthorized drug consumption unnecessary. Catheterization of the portal vein across the adjacent edge of the liver seals the punctured hole reliably with a U-shaped suture which prevents bleeding into the abdomen and secures the catheter in place. PMID- 20020661 TI - [Arterial embolization as palliation for inoperable renal carcinoma]. AB - Embolization of the renal artery was performed as a palliative measure in 93 inoperable patients at the Center's Clinic in 1991-2000. Radio-endovascular intervention was indicated in cases of bleeding (19, 20.5%), case-history macrohematuria with high risk of relapse (54, 58%) and drug-controlled pain (20, 21.5%). Bleeding was suspended for 3-16 days (average 7.5 +/- 4) after occlusion of the artery in 16 out of 19 patients and decreased significantly in 3 (16%). There was no bleeding after embolization among macrohematuria patients. Pain syndrome was abated in 70% while post-intervention complications occurred in 13%: they were transitory in 6.5% and--fatal in another 6.5%. Catheter-assisted arterial embolization is a fairly safe and efficient means of arresting bleeding from inoperable renal kidney coupled with marked analgesic effect. PMID- 20020662 TI - [Final results of cooperative studies of dicarbamine used as a hemoprotector in combination chemotherapy for cancer]. PMID- 20020663 TI - [The use of computed X-ray tomography in laryngeal and laryngopharyngeal cancer: history and current trends]. PMID- 20020664 TI - [Thyroid cancer: current understanding of etiology, pathogenesis and diagnosis]. PMID- 20020665 TI - [Current therapy for dissiminated soft-tissue sarcoma in Russia]. PMID- 20020666 TI - New drug development. PMID- 20020667 TI - PARP inhibition in breast cancer. PMID- 20020668 TI - Current status of acute myeloid leukemia treatment in the elderly. PMID- 20020669 TI - New antiplatelet agents. PMID- 20020670 TI - Lessons learned from the development of kinase inhibitors. PMID- 20020671 TI - HER2-neu positivity in patients with small and node-negative breast cancer (pT1a,b,N0,M0): a high risk group? AB - Human epidermal growth factor receptor 2 (HER2-neu) is an important prognostic factor associated with worsened disease-free survival and overall survival in breast cancer patients; however, the prognosis of T1a,b,N0,M0 HER2-neu positive breast cancer has not been clearly determined. Trastuzumab has become a critical component of the treatment of patients with HER2-neu positive tumors, but the effect of treatment in patients with small tumors and node-negative disease has not been evaluated in clinical trials. Current guidelines have category 3 recommendations to consider the use of adjuvant trastuzumab in women with node negative tumors that are 0.6-1.0 cm, and state that physicians should balance the risks associated with the known trastuzumab toxicities and the uncertain benefits that may exist with such therapy in this group of patients. The available data regarding the prognosis of patients with T1a,b,N0,M0 HER2-neu positive breast cancer is very limited. The purpose of this manuscript is to review the available literature in order to evaluate whether this group of patients represents a high risk group. Retrospective studies suggest that HER2-neu status is a powerful independent prognostic factor in T1a,b node-negative breast cancer. We believe that prospective studies and randomized clinical trials are strongly needed to clearly assess the impact of HER2-neu positivity in node-negative and subcentimeter tumors, and to determine if this group of patients can benefit from adjuvant trastuzumab treatment. PMID- 20020672 TI - Incidence and management of asparaginase-associated adverse events in patients with acute lymphoblastic leukemia. AB - Asparaginase is an enzyme that breaks down extracellular asparagine into aspartic acid and ammonia. Depletion of extracellular asparagine inhibits the growth of lymphocytic leukemic cells. Unlike normal cells, lymphoblasts lack the enzyme to synthesize asparagine and therefore rely on an exogenous source of this amino acid to maintain cellular protein synthesis. Asparagine depletion results in nutritional deprivation, inhibition of protein synthesis, and subsequent apoptotic cell death in lymphoblasts. Asparaginase therapy is an essential component of the treatment protocol for acute lymphoblastic leukemia. The effect of asparaginase on protein synthesis may result in a number of toxicities, including thrombosis, pancreatitis, hyperglycemia, and hepatotoxicity. This review discusses the incidence of asparaginase-related adverse events, compares available asparaginase formulations with respect to the emergence of certain toxicities, and considers management strategies for these toxicities in patients with acute lymphoblastic leukemia. PMID- 20020673 TI - Reducing in fl uenza spreading over the airline network. AB - Disease spreading through human travel networks has been a topic of great interest in recent years, such as with swine in fl uenza or SARS pandemics. Most studies have proposed removing highly connected nodes (hubs) to control spreading. Here, we test alternative strategies using edge removal ( fl ight cancellation) for spreading over the airline network. Flight cancellation was more ef fi cient than shutting down whole airports: spreading took 81% longer if solely selected fl ights were removed, compared to a 52% reduction when entire airports were shutdown, affecting the same number of fl ights. PMID- 20020674 TI - First estimation of direct H1N1pdm virulence: From reported non consolidated data from Mauritius and New Caledonia. AB - We provide rough estimates of direct lethality from ARDS due to H1N1pdm from two independent sources of data, one from New Caledonia where 30,000 infections are assumed to have occured and 3 deaths reported to be attributable directly to the pandemic virus. Another source is Mauritius where 70,000 infections are estimated to have occured, and 7 reported death from ARDS (5 of them are currently confirmed). These surveillance data allows for first estimation of direct lethality due to H1N1pdm to be 1 per 10,000 infections, about 100 times more than regular seasonal influenza. PMID- 20020675 TI - Fabrication and characterization of DNA arrays prepared on carbon-on-metal substrates. AB - Carbon-on-metal substrates consist of a surface plasmon-conducting metal substrate with a thin amorphous carbon overlayer. Recently, oligonucleotide arrays were in situ synthesized on carbon-on-gold substrates, and DNA hybridization experiments were monitored with surface plasmon resonance (SPR) imaging. We describe here the thorough characterization of these substrates and arrays as they progress through the fabrication process. Two surface plasmon conducting metals, gold and silver, were utilized in the carbon-on-metal substrate preparation and their SPR responses compared. Oligonucleotide arrays synthesized on the carbon-on-metal substrates were analyzed with fluorescence- and SPR-based measurements. The stability of the carbon-on-metal substrates when exposed to prolonged incubations and/or serial hybridizations was also determined. PMID- 20020676 TI - Photoreactions of mercury in surface ocean water: gross reaction kinetics and possible pathways. AB - We present pseudofirst order rate constants for gross photoreduction and gross photooxidation of mercury in surface water from the open Atlantic Ocean, determined under controlled laboratory conditions. Experiments using both unfiltered and filtered ocean water were carried out to characterize the importance of microbes and colloids on reaction kinetics. Results indicate that reduction and oxidation of mercury in ocean water does not follow a simple two species reversible reaction pathway. We suggest two possible redox pathways that reproduce the pattern of dissolved gaseous mercury (DGM) concentrations observed in our laboratory experiments, and evaluate them using a controlled outdoor experiment. In both proposed pathways Hg(0), the major constituent of DGM, is converted to an unidentified oxidized species that is different from the reducible form present initially. This reaction step plays a major role in the net formation of DGM in our experiments. Our results represent new quantitative information about the gross reaction kinetics for both reduction and oxidation of mercury in open ocean surface water. Pseudofirst order rate constants for reduction reactions that form DGM were determined to be in the range of 0.15-0.93 h(-1) and pseudofirst order rate constants for oxidation of Hg(0) to be in the range of 0.4-1.9 h(-1). Microbes and colloids did not appreciably influence the reduction and oxidation kinetics. PMID- 20020677 TI - Evidence of compound-dependent hydrodynamic and mechanical transverse dispersion by multitracer laboratory experiments. AB - Mass transfer, mixing, and therefore reaction rates during transport of solutes in porous media strongly depend on dispersion and diffusion. In particular, transverse mixing is a significant mechanism controlling natural attenuation of contaminant plumes in groundwater. The aim of the present study is to gain a deeper understanding of vertical transverse dispersive mixing of reaction partners in saturated porous media. Multitracer laboratory experiments in a quasi two-dimensional tank filled with glass beads were conducted and transverse dispersion coefficients were determined from high-resolution vertical concentration profiles. We investigated the behavior of conservative tracers (i.e., fluorescein, dissolved oxygen, and bromide), with different aqueous diffusion coefficients, in a range of grain-related Peclet numbers between 1 and 562. The experimental results do not agree with the classical linear parametric model of hydrodynamic dispersion, in which the transverse component is approximated as the sum of pore diffusion and a compound-independent mechanical dispersion term. The outcome of the multitracer experiments clearly indicates a nonlinear relation between the dispersion coefficient and the average linear velocity. More importantly, we show that transverse mechanical dispersion depends on the diffusion coefficient of the compound, at least at the experimental bench scale. This result has to be considered in reactive-transport models, because the typical assumption that two reactants with different aqueous diffusive properties are characterized by the same dispersive behavior does not hold anymore. PMID- 20020678 TI - Identifying health impacts of exposure to copper using transcriptomics and metabolomics in a fish model. AB - Copper (Cu) is a micronutrient essential for the biochemical functioning of numerous processes in vertebrates but is also often present in the aquatic environment at concentrations able to cause adverse health effects in aquatic organisms. This study investigated the signaling pathways mediating the effects of exposure to Cu using a toxicogenomic approach in a fish model, the stickleback ( Gasterosteus aculeatus ). Freshwater-acclimated male fish were exposed via the water to Cu, including at environmentally relevant concentrations (3.2-128 microg of Cu/L for 4 days), and the biological responses explored through analyses of the hepatic transcriptome and metabolome and phenotypic end points, including assessment of DNA damage in blood cells. The Cu exposures resulted in DNA strand breaks in blood cells at all exposure concentrations and alterations in hepatic gene expression and metabolite concentrations in a concentration-dependent manner (from 10 microg of Cu/L). Genes associated with the cholesterol biosynthesis pathway were significantly over-represented and consistently down-regulated (at 128 microg of Cu/L), similar to that occurring in a mouse model for Wilson's disease. Additionally, inductions in metallothionein and catalase were also observed. The concentrations of NAD(+) and lactate increased significantly with the Cu exposure, consistent with a shift toward anaerobic metabolism, and these aligned closely with changes observed in gene expression. The pathways of Cu toxicity identified in our study support the conserved mechanisms of Cu toxicity from lower vertebrates to mammals, provide novel insights into the deleterious effects of Cu in fish, and further demonstrate the utility of fish as environmental sentinels for chemical impacts on both environmental and human health. PMID- 20020679 TI - Deposition of mercury species in the Ny-Alesund area (79 degrees N) and their transfer during snowmelt. AB - Arctic snowpacks are often considered as temporary reservoirs for atmospheric mercury (Hg) deposited during springtime deposition events (AMDEs). The fate of deposited species is of utmost importance because melt leads to the transfer of contaminants to snowmelt-fed ecosystems. Here, we examined the deposition, fate, and transfer of mercury species (total Hg (THg) and methylmercury (MeHg)) in an arctic environment from the beginning of mass deposition of Hg during AMDEs to the full melt of the snow. Following these events, important amounts of THg were deposited onto the snow surface with concentrations reaching 373 ng.L(-1) and estimated deposition fluxes of 200-2160 ng.m(-2). Most of the deposited Hg was re emitted to the atmosphere via photochemical reactions. However, a fraction remained stored in the snow and we estimated that the spring melt contributed to an input of 1.5-3.6 kg.year(-1) of THg to the fjord (i.e., 8-21% of the fjord's THg content). A monitoring of MeHg in snow using a new technique (DGT sensors) is also presented. PMID- 20020680 TI - A small molecule-regulated guanine nucleotide exchange factor. AB - Selective, pharmacological agents are attractive tools for studying signal transduction because they allow rapid, reversible, and dose-dependent control over intracellular protein function. However, for many targets the identification of potent and selective small molecule agonists and antagonists is a formidable challenge. An attractive strategy for circumventing this problem is to engineer a protein of interest to be sensitive to a pharmacological agent of choice. Here, we report a chemical genetic method for regulating the catalytic activity of signaling enzymes with a small molecule. This approach uses the interaction of the antiapoptotic protein Bcl-xL and a BH3 peptide as an autoinhibitory switch that can be controlled with a small molecule. We applied this strategy to the guanine nucleotide exchange factor Intersectin, which is a selective activator of the GTPase Cdc42. Replacing Intersectin's regulatory domains with the BH3 peptide/Bcl-xL binding module generated a panel of synthetic GEF constructs that can be activated with a competitive ligand. Importantly, the nucleotide exchange activities of these synthetic Intersectin constructs can be controlled in a rapid and dose-dependent manner. The modular nature of this strategy should make it useful for engineering other enzymes involved in signal transduction. PMID- 20020681 TI - Dual redox responsive assemblies formed from diselenide block copolymers. AB - A block copolymer with diselenide bonds in the polymer backbone was reported. This block copolymer was capable of forming micellar aggregates that were responsive to redox stimuli. Compared with other redox responsive aggregates, this type of diselenide-containing block copolymer aggregates could be responsive to both oxidants and reductants even in a solution with a very low concentration under mild conditions. PMID- 20020682 TI - Practical asymmetric conjugate alkynylation of Meldrum's acid-derived acceptors: access to chiral beta-alkynyl acids. AB - The enantioselective conjugate addition of alkynyl nucleophiles has been a long standing challenge in synthetic chemistry. This paper describes a highly practical asymmetric conjugate alkynylation of Meldrum's acid-derived acceptors using cinchonidine (<$100/kg) as the chiral mediator. The process provides practical access to chiral beta-alkynyl acids. Noteworthy attributes of the method are its broad scope, high functional-group compatibility, and ease of scalability. PMID- 20020683 TI - Changes in the volatile compound production of fermentations made from musts with increasing grape content. AB - Wine is a complex consumer product produced predominately by the action of yeast upon grape juice. Model must systems have proven to be ideal for studies into the effects of fermentation conditions on the production of certain wine volatiles. To clarify the contribution of grape juice to the production of wine volatiles, we have employed a model must system spiked with increasing amounts of grape juice (Riesling or Cabernet Sauvignon). The resulting fermented wines were analyzed by SPME-GC-MS and the data obtained grouped using ANOVA and cluster analyses to reveal those compounds that varied in concentration with reproducible trends relative to juice concentration. Such grouping highlights those compounds that are grape-dependent or for which production is modulated by grape composition. In some cases, increasing the proportion of grape juice in the fermentations stimulated the production of certain esters to levels between 2- and 140-fold higher than those seen in fermentations made with model grape juice media alone. The identification of the grape components responsible for the increased production of these wine volatiles will have implications for the impact of grape production and enology on wine flavor and aroma. PMID- 20020684 TI - In vitro bile-acid binding and fermentation of high, medium, and low molecular weight beta-glucan. AB - The impact of beta-glucan molecular weight (MW) on in vitro bile-acid binding and in vitro fermentation with human fecal flora was evaluated. beta-Glucan extracted from oat line 'N979-5-4' was treated with lichenase (1,3-1,4-beta-D-glucanase) to yield high (6.87x10(5) g/mol), medium (3.71x10(5) g/mol), and low (1.56x10(5) g/mol) MW fractions. The low MW beta-glucan bound more bile acid than did the high MW beta-glucan (p<0.05). If the positive control, cholestyramine, was considered to bind bile acid at 100%, the relative bile-acid binding of the original oat flour and the extracted beta-glucan with high, medium, and low MW was 15, 27, 24, and 21%, respectively. Significant effects of high, medium, and low MW beta-glucans on total SCFA were observed compared to the blank without substrate (p<0.05). There were no differences in pH changes and total gas production among high, medium, and low MW beta-glucans, and lactulose. The low MW beta-glucan produced greater amounts of SCFA than the high MW after 24 h of fermentation. Among the major SCFA, more propionate was produced from all MW fractions of extracted beta-glucans than from lactulose. In vitro fermentation of extracted beta-glucan fractions with different MW lowered pH and produced SCFA, providing potential biological function. PMID- 20020685 TI - New platform of biosensors for prescreening of pesticide residues to support laboratory analyses. AB - Millions of tons of pesticides are applied worldwide annually in agriculture. Among them, herbicides such as triazines and ureas, originating from agricultural runoff, can contaminate soils and surface and ground waters with severe toxic effects on humans. Nowadays, different analytical techniques are available for the detection of these chemicals; however, most of them are expensive and time consuming, especially in the case of routine analyses. For this reason, on the basis of results collected through many years of experience in the field of photosynthetic organisms, we designed a biosensor platform intended for the easy, low-cost, and fast prescreening of photosynthetic herbicides. The platform combines the possibilities of amperometric and optical transduction systems, which have proven to be highly sensitive (limits of detection = 10(-10)-10(-8) M). The use of genetically modified algae strengthens the power of the platform, allowing different subclasses of herbicides to be recognized. The system has been validated for the analysis of environmental water and is proposed to support laboratories involved in the control of water pollution. PMID- 20020686 TI - Superior contact for single-molecule conductance: electronic coupling of thiolate and isothiocyanate on Pt, Pd, and Au. AB - One of the critical issues for the realization of molecular electronics is the development of ideal molecule-electrode contacts that render efficient charge transportation and thus attenuate the unwanted voltage drop and power loss. The conductance at the single-molecule level has long been expected to be correlated strongly with the electrode materials. However, other than gold, systematic studies of a homologous series of molecules to extract the headgroup-metal contact conductance (G(n=0)) have not been reported. Carefully examined herein are the conductances of alkanedithiols anchored onto electrode materials of Au and Pt as well as the conductances of alkanediisothiocyanates on Au, Pd, and Pt by utilizing the method of STM-BJ (scanning tunneling microscopy break junction). In comparison with Au substrate, Pd and Pt are group 10 elements with stronger d orbital characteristics, and larger local density of states near the Fermi level. The model compounds, SCN(CH(2))(n)NCS (n = 4, 6, and 8), are studied because the isothiocyanate (-NCS) headgroup is a versatile ligand for organometallics, an emerging class of molecular wires, and can bind to substrates of noble metals to complete a metal-molecule-metal configuration for external I-V measurements. Also studied include alkanedithiols, one of the most scrutinized systems in the field of single-molecule conductance. The results show that the conductance for single molecules bridged between a pair of Pt electrodes is about 3.5-fold superior to those between Au electrodes. On all electrode materials, observed are two sets of conductance values, with the smaller set being 1 order of magnitude less conductive. These findings are ascribed to the degree of electronic coupling between the headgroup and the electrode. PMID- 20020687 TI - Recent trends and advances in berry health benefits research. AB - Recent advances have been made in our scientific understanding of how berries promote human health and prevent chronic illnesses such as some cancers, heart disease, and neurodegenerative diseases. Cancer is rapidly overtaking heart disease as the number one killer disease in developed countries, and this phenomenon is coupled with a growing aging population and concomitant age-related diseases. Therefore, it is not surprising that consumers are turning toward foods with medicinal properties as promising dietary interventions for disease prevention and health maintenance. Among fruits, berries of all colors have emerged as champions with substantial research data supporting their abilities to positively affect multiple disease states. Apart from several essential dietary components found in berries, such as vitamins, minerals, and fiber, berries also contain numerous bioactives that provide health benefits that extend beyond basic nutrition. Berry bioactives encompass a wide diversity of phytochemicals (phytonutrients) ranging from fat-soluble/lipophilic to water-soluble/hydrophilic compounds. Recent research from laboratories across the globe has provided useful insights into the biological effects and underlying mechanisms of actions resulting from eating berries. The cluster of papers included here represents a cross section of topics discussed at the 2009 International Berry Health Benefits Symposium. Together, these papers provide valuable insight into recent research trends and advances made into evaluating the various health benefits that may result from the consumption of berries and their derived products. PMID- 20020688 TI - Subcritical solvent extraction of procyanidins from dried red grape pomace. AB - Procyanidins in dried Sunbelt ( Vitis labrusca L.) red grape pomace were extracted using accelerated solvent extraction (ASE) with pressure (6.8 MPa), one extraction cycle, and temperature (40, 60, 80, 100, 120, and 140 degrees C). Six ethanol/water solvents (0, 10, 30, 50, 70, and 90%, v/v) were compared to conventional extraction with acetone/water/acetic acid (70:29.5:0.5, v/v/v). Procyanidins in the extracts were identified by HPLC-ESI-MS/MS and contained degrees of polymerization (DP) of 1-5 (monomers through pentamers) and polymers (DP > 10). Generally, 50% ethanol/water (v/v) extracted more total procyanidins than other ethanol/water compositions, and contained up to 115% of total procyanidins extracted by the acetone-based conventional solvent. Additionally, 50% ethanol/water (v/v) extracted 205, 221, and 113% more epicatechin, catechin, and dimers, respectively, than conventional extraction. Results indicated greater extraction of low oligomeric procyanidins using 50% ethanol/water (v/v) solvent between 80 and 140 degrees C. PMID- 20020689 TI - Proximate and polyphenolic characterization of cranberry pomace. AB - The proximate composition and identification and quantification of polyphenolic compounds in dried cranberry pomace were determined. Proximate analysis was conducted based on AOAC methods for moisture, protein, fat, dietary fiber, and ash. Other carbohydrates were determined by the difference method. Polyphenolic compounds were identified and quantified by HPLC-ESI-MS. The composition of dried cranberry pomace was 4.5% moisture, 2.2% protein, 12.0% fat, 65.5% insoluble fiber, 5.7% soluble fiber, 8.4% other carbohydrates, 1.1% ash, and 0.6% total polyphenolics. It contained six anthocyanins (111.5 mg/100 g of DW) including derivatives of cyanidin and peonidin. Thirteen flavonols were identified (358.4 mg/100 g of DW), and the aglycones myricetin (55.6 mg/100 g of DW) and quercetin (146.2 mg/100 g of DW) were the most prominent. Procyanidins with degrees of polymerization (DP) of 1-6 were identified (167.3 mg/100 g of DW), the most abundant being an A-type of DP2 (82.6 mg/100 g of DW). PMID- 20020690 TI - Polyphenolic composition and antioxidant capacity of extruded cranberry pomace. AB - Cranberry pomace was mixed with corn starch in various ratios (30:70, 40:60, 50:50 pomace/corn starch DW) and extruded using a twin-screw extruder at three temperatures (150, 170, 190 degrees C) and two screw speeds (150, 200 rpm). Changes in the anthocyanin, flavonol, and procyanidin contents due to extrusion were determined by HPLC. Antioxidant capacity of the extrudates was determined using oxygen radical absorbance capacity (ORAC). Anthocyanin retention was dependent upon barrel temperature and percent pomace. The highest retention was observed at 150 degrees C and 30% pomace. Flavonols increased by 30-34% upon extrusion compared to an unextruded control. ORAC values increased upon extrusion at 170 and 190 degrees C. An increase in DP1 and DP2 procyanidins was also observed; however, a decrease was observed in DP4-DP9 oligomers. These data suggest that extrusion alters the polyphenolic distribution of cranberry pomace and has application in the nutraceutical industry as a means of improving the functionality of this coproduct. PMID- 20020691 TI - Plasmon-enhanced upconversion in single NaYF4:Yb3+/Er3+ codoped nanocrystals. AB - In this Letter we report the plasmon-enhanced upconversion in single NaYF(4) nanocrystals codoped with Yb(3+)/Er(3+). Single nanocrystals and gold nanospheres are investigated and assembled in a combined confocal and atomic force microscope setup. The nanocrystals show strong upconversion emission in the green and red under excitation with a continuous wave laser in the near-infrared at 973 nm. By the use of the atomic force microscope, we couple single nanocrystals with gold spheres (30 and 60 nm in diameter) to obtain enhanced upconversion emission. An overall enhancement factor of 3.8 is reached. A comparison of time-resolved measurements on the bare nanocrystal and the coupled nanocrystal-gold sphere systems unveil that faster excitation as well as faster emission occurs in the nanocrystals. PMID- 20020692 TI - Determination of the provenance of cocoa by soil protolith ages and assessment of anthropogenic lead contamination by pb/nd and lead isotope ratios. AB - The Pb contents of chocolate and the products it flavors are among the highest of all commonly consumed substances. Others have shown that this Pb is acquired by cocoa beans after harvesting and is concentrated in their shells, portions of which are ground up with the cotyledons during processing. It is shown here that the shells also contain the lanthanides Nd and Sm, which they appear to take up more slowly than Pb when dried on bare soil. Consideration of Pb/Nd ratios, model Sm-Nd ages and the isotope ratios of Pb and Sr indicates that, in the absence of contamination, the relationship between Pb and Nd in shells is y = 13.1x(-0.383), where x is the Nd concentration in microg/kg and y is the Pb/Nd ratio. For cocoa powders, the relationship is y = 114x(-0.988). Samples that plot above these curves are probably contaminated. Model ages indicate where the cocoa of cocoa powders is grown, and these same considerations point to African samples being uncontaminated but samples from Asia containing 50% anthropogenic Pb of Australian origin. No measurable Pb contamination occurs during the transport of beans and the manufacture of chocolate. PMID- 20020693 TI - Phenolic compounds and furanic derivatives in the characterization and quality control of Brandy de Jerez. AB - This article shows the results obtained in the study of the extraction profiles from oak wood to distillate of several compounds, low molecular weight phenolics, and furanic derivatives, and the relationship of their contents with those found in commercial sherry brandies and other aged distillates of different geographical origin, in order to research the utility of these analytical variables for explaining the highly specific character of Brandy de Jerez. Using multivariate statistic techniques, the aging system (static by anadas, or dynamic, well known as Soleras y Criaderas) has been confirmed as having a great influence on the analytical profile of aged distillates (discrimination is up to 100%). Differences between commercial brandies and those aged experimentally of equivalent average age have also been confirmed. The Solera Gran Reserva Brandies de Jerez show a clear differentiation from the rest of the distillates of different origin (discrimination is up to 80%), indicating their highly specific character. PMID- 20020694 TI - Unsymmetrical tripodal ligand for lanthanide complexation: structural, thermodynamic, and photophysical studies. AB - Two tridentate and one bidentate binding strands have been anchored on a carbon atom to provide a new unsymmetrical tripodal ligand L for Ln(III) coordination. The ligand itself adopts a single conformation in solution stabilized by intramolecular hydrogen bonds evidenced in the solid state. The reaction of L with trivalent lanthanides provides different coordination complexes depending on the metal/ligand ratio. The speciation studies with selected lanthanides were performed in solution by means of NMR, ESMS, and spectrophotometric titrations. Differences in coordination properties along the lanthanide series were evidenced and may be associated with the changes in the ionic size. However, thermodynamic stability constants for the species of the same stoichiometry do not significantly vary. In addition, the structure of the dinuclear complex [Eu(2)L(2)](6+) has been elucidated in the solid state, where the complex crystallizes predominantly as an M-isomer. The crystal structure shows the coordination of two different ligands to each europium cation through tridentate strands, and the europium nine-coordinate sphere is completed with three solvent molecules. Finally, the results of photophysical investigations of [Eu(2)L(2)](6+) are in close agreement with the structural parameters determined by crystallography. PMID- 20020695 TI - Fractionation of Gallotannins from mango (Mangifera indica L.) kernels by high speed counter-current chromatography and determination of their antibacterial activity. AB - High-speed counter-current chromatography was applied to the separation of gallotannins from mango (Mangifera indica L.) kernels. The kernels were defatted and subsequently extracted with aqueous acetone [80% (v/v)]. The crude extract was purified by being partitioned against ethyl acetate. A hexane/ethyl acetate/methanol/water solvent system [0.5:5:1:5 (v/v/v/v)] was used in the head to-tail mode to elute tannins according to their degree of galloylation (tetra-O galloylglucose to deca-O-galloylglucose). The compounds were characterized using liquid chromatography and mass spectrometry in the negative ionization mode. Purities ranged from 72% (tetra-O-galloylglucose) to 100% (octa-O galloylglucose). The iron binding capacity of gallotannins was dependent on the number of galloyl groups in the molecule, with a larger capacity at lower degrees of galloylation. The minimum inhibitory concentration against Bacillus subtilis did not change among the different gallotannins tested and was in the range of 0.05-0.1 g/L in Luria-Bertani broth but up to 20 times higher in media containing more iron and divalent cations. PMID- 20020696 TI - Facile synthesis of 2,6-disubstituted benzobisthiazoles: functional monomers for the design of organic semiconductors. AB - The synthesis of several synthetically useful 2,6-disubstituted benzobisthiazoles is described. The method is based on the Lewis acid-catalyzed ring-closing reaction between substituted orthoesters and diamino benzene dithiol. The resulting benzobisthiazoles are obtained cleanly and in good yields. These materials are of interest for the development of new organic semiconductors. PMID- 20020697 TI - Preparation of tethered aldehyde ynoates and ynones by ring fragmentation of cyclic gamma-oxy-beta-hydroxy-alpha-diazo carbonyls. AB - Cyclic gamma-oxy-beta-hydroxy-alpha-diazo carbonyls undergo Lewis acid induced ring fragmentation to provide either ynoates or ynones tethered to an aldehyde, ketone, or ester. The fragmentation precursors are convenient to prepare by adding lithiated alpha-diazo carbonyls to alpha-oxy ketones. The fragmentation appears general and provides a variety of functional group-rich products in good to excellent yield. PMID- 20020698 TI - Acute and repeated dose (28 days) oral safety studies of an alkoxyglycerol extract from shark liver oil in rats. AB - Shark liver oil has been used for over 50 years as both a therapeutic and preventive agent. The active ingredients in shark liver oil have been found to be a group of ether-linked glycerols known as alkoxyglycerols. Despite its popularity, there is little published toxicology data on alkoxyglycerols. The toxicity of a supercritical fluid extract of shark liver oil (AKG-1 extract) has been evaluated in acute and repeated dose (28 days) oral toxicity studies in rats at doses of 200 and 100 times the maximum recommended dose by supplement manufacturers in humans, respectively. The AKG-1 extract administered in a single oral gavage dose of 2000 mg kg(-1) of body weight resulted in no adverse events or mortality. The AKG-1 extract administered as a daily dose of 1000 mg kg(-1) of body weight for 28 days by gavage resulted in no adverse effects or mortality. For both studies, no abnormal clinical signs, behavioral changes, body weight changes, or change in food and water consumption occurred. There were no changes in hematological and serum chemistry values, organ weights, or gross or histological characteristics. It is concluded that the AKG-1 extract is well tolerated in rats at an acute dose of 2000 mg kg(-1) and at a subchronic (28 days) dose of 1000 mg kg(-1). PMID- 20020699 TI - Multiresidue analysis of 50 pesticides in grape, pomegranate, and mango by gas chromatography-ion trap mass spectrometry. AB - A selective and sensitive multiresidue analysis method is reported for simultaneous determination of 50 pesticides of different chemical classes in three commercially important fruits of different nature viz. grape, pomegranate, and mango. The sample preparation method involves extraction of a 10 g sample with 10 mL of ethyl acetate; cleanup by dispersive solid phase extraction with primary secondary amine (PSA, 25 mg) for grape and PSA + graphitized carbon black (25 + 5 mg) for pomegranate and mango; and determination by gas chromatography ion trap mass spectrometry through multiple reaction monitoring (MRM). Sample preparation under acidified (pH 4) and cold (<4 degrees C) conditions, use of PTV large volume injection (20 microL) through multibaffled liner and chromatographic separation on a short 10 m VF-5MS capillary column gave a satisfactory response for all of the analytes including relatively unstable compounds such as captan, captafol, folpet, endrine, and iprodione within 31.8 min. The limit of quantification (LOQ) of most of the compounds was 0 and DeltaS > 0, in sharp contrast to first hydration processes in nonpolar media. The present results represent the first theoretical attempt to rationalize the large amount of experimental data on hydration and aggregation of Chla in aqueous organic media that have been accumulated over the past four decades. The data stress, in particular, the key role of Chla dihydrates, a point that has been the object of intense debate in the literature. Clearly, dihydrates are found to be more stable than monohydrates owing to a particular structure in which cooperative interactions occur between the water molecules and Chla. The calculations also explain the irregular behavior observed for Chla in aqueous THF or pyridine: In these media, Chla remains basically unhydrated because the Chla-solvent adducts are stabilized by strong dispersion interactions. PMID- 20020704 TI - Selectivity in the reaction of triplet phenyl cations. AB - A DFT study of the reaction of phenyl cation and some 4-substituted derivatives (cyano, methyl, methoxy, amino) with a pi nucleophile (ethylene) as well as with representative n nucleophiles (NH(3), MeOH, and MeCN) reveals a multiform behavior depending on both the cation multiplicity and the trap used. A straightforward addition takes place with the singlet (pi(6)sigma(0) structure) both with ethylene, where a spiro[2,5]octa-4,7-dienyl (phenonium) cation is formed, and with n nucleophiles, where the corresponding onium cations result. On the contrary, with the triplet (pi(5)sigma(1) structure) the reaction depends on the nature of the nucleophile, as indicated by MO correlation diagrams. Thus, with ethylene a bonding interaction occurs between the singly occupied sigma(sp(2)) orbital of the cation and the alkene pi orbital and leads to a planar distonic diradical cation. On the contrary, no addition takes place with n nucleophiles, which interact only with the phenyl cations pi MO, leading to weakly bonded, face-to-face complexes. An electron-withdrawing substituent such as CN allows the formation of a stabilized adduct cation also from the triplet, but only with a good nucleophile, such as ammonia. The spin-dependent dichotomy in the chemical behavior rationalizes recent experimental findings and fits with the prediction formulated by Taft 45 years ago. The unusual combination of a carbocation nature and of triplet multiplicity originates the peculiar chemistry of phenyl cations that appear to be promising intermediates in synthesis. PMID- 20020705 TI - Theoretical study on the mechanism of the reaction of CF(3)S with NO(2). AB - The singlet potential energy surface for the CF3S + NO2 reaction has been theoretically investigated using the B3LYP/6-311+G(3df) level of theory. The geometries, vibrational frequencies, and zero-point energies of all stationary points involved in the title reaction have been examined. More accurate energies of stationary points were obtained using ab initio G3//B3LYP and CBS-QB3 composite methods. The results show that the initial addition of CF3S with NO2 leads to CF3SNO2 or CF3SONO intermediates, which are formed without an electronic barrier. CF3SNO2 can easily isomerizes to CF3SONO, while CF3SONO readily isomerizes to CF3S(O)NO or dissociates to CF3SO + NO, which are the major products of the title reaction. Reaction channels leading to the formation the CF3O + SNO and CF2S + FNO2 products are highly improbable processes due to high energy barriers involved. We have also computed heats of formation for CF3SNO2, CF3SONO, and CF3S(O)NO intermediates. It was found that the most stable is the cis-perpendicular form of CF3SONO isomer with DeltaH(f,0)0 = -243.6 kcal mol-1. PMID- 20020706 TI - Role of pi-conjugation in influencing the magnetic interactions in dinitrenes: a broken-symmetry approach. AB - In the present study, we investigated the magnetic interactions of some dinitrenes by employing the broken symmetry-unrestricted density functional theoretical (BS-UDFT) approach along with the use of three basis sets. The magnetic coupling parameter (J) has been calculated, and thereby the magnetic character of the molecule and the strength of magnetic interaction are explored for these molecules. The exchange coupling parameters for the corresponding unconjugated systems are also calculated to see the role of pi-conjugation. Our results suggest that a strong antiferromagnetic interaction exists in conjugated dinitrenes, and the strength of magnetic interaction decreases with increase in spacer length. For the unconjugated dinitrenes, the nature of magnetic interaction reduced appreciably and becomes weakly antiferromagnetic. The singlet triplet energy gap for each system is also calculated. For the conjugated systems, it is observed that the singlet states are more stable than the triplet states, whereas for the unconjugated systems, the relative stability of the singlet state reduces to a considerable extent. This discrepancy of results for the conjugated and unconjugated dinitrenes can be attributed to the effect of pi conjugation, and the results can be well explained by this effect. PMID- 20020707 TI - New dimeric stilbenoids from fungal-challenged peanut ( Arachis hypogaea) seeds. AB - The peanut plant can resist fungal attacks by producing stilbene-derived phytoalexins. Once understood, such a natural phytoalexin-based mechanism of peanut resistance could be potentially manipulated to obtain fungal-resistant peanut breeding lines. Several simple stilbenoid phytoalexins from peanuts have been reported. However, more complex stilbenoid derivatives such as those that have been reported from other sources and considered important factors in plant defense have not been found in peanuts. The purpose of this research was to isolate and characterize further new oligomeric peanut stilbenoids that may act as phytoalexins. Two new prenylated stilbene dimers named arahypin-6 (3) and arahypin-7 (4) have been isolated from wounded peanut seeds challenged by an Aspergillus caelatus strain. The structures of these new putative phytoalexins were determined by analysis of NMR, MS, and UV spectroscopic data. Together with other known peanut stilbenoids that were also produced in the challenged seeds, these new compounds may play a defensive role against invasive fungi. PMID- 20020708 TI - Electron affinity of trans-2-C4F8 from electron attachment-detachment kinetics. AB - Electron attachment and detachment kinetics of 2-C(4)F(8) were studied over the temperature range 298-487 K with a flowing-afterglow Langmuir-probe apparatus. Only parent anions were formed in the attachment process throughout this temperature range. At the highest temperatures, thermal electron detachment of the parent anions is important. Analysis of the 2-C(4)F(8) gas showed an 82/18 mixture of trans/cis isomers. The kinetic data at the higher temperatures were used to determine the electron affinity EA(trans-2-C(4)F(8)) = 0.79 +/- 0.06 eV after making some reasonable assumptions. The same quantity was calculated using the G3(MP2) compound method, yielding 0.74 eV. The kinetic data were not sufficient to establish a reliable value for EA(cis-2-C(4)F(8)), but G3(MP2) calculations give a value 0.017 eV greater than that for trans-2-C(4)F(8). MP2 and density functional theory were used to study the structural properties of the neutral and anion isomers. PMID- 20020709 TI - Wavelength-dependent photodissociation of benzoic acid monomer in alpha C-O fission. AB - In concert with the latest laser-induced fluorescence (LIF) experiment [Wei et al. J. Phys. Chem. A 2008, 112, 4727], we investigated the photodissociation mechanics of the benzoic acid monomer (BAM) with alpha C-O fission by means of state-of-the-art ab initio calculations. Complete active space self-consistent field (CASSCF) and multireference CASSCF second-order perturbation theory (MSCASPT2) calculations were performed on the ground and a number of low-lying excited states of BAM. Our calculations indicated that alpha C-O fission from the S(1) state is in competition with the fission from the T(2) state upon the 266 284 nm wavelength photon. This differs from the conclusion of the previous theoretical investigation and clarified the vague experimental conclusion made earlier. According to our calculations, alpha C-O fission mainly occurs at the T(2) state upon photoexcitation at 284-294 nm, and the photon with a wavelength longer than 294 nm is unable to present the alpha C-O fission. This conclusion agrees with the LIF experimental observation. PMID- 20020710 TI - Novel KCNQ2/Q3 agonists as potential therapeutics for epilepsy and neuropathic pain. AB - Current drugs for the treatment of seizure disorders, although effective in many patients, still suffer from a number of failures and are not effective in some forms of resistant epilepsies. Historically, many of these drugs have multiple mechanisms of action including calcium and sodium channel blockade as well as GABAergic activity and thus a number of associated side effects. Modulation of the M-current through opening of KCNQ channels has been proposed as a way to attenuate neuroexcitability and have a therapeutic benefit for the treatment of seizure disorders. Therefore, as part of our program to identify new treatments for epilepsy, we set out to identify agonists of KCNQ channels. High throughput screening of our corporate collection led to the identification of 1, adamantane 1-carboxylic acid (3-methyl-3H-benzothiazol-2-ylidine) hydrazide, a potent KCNQ2/Q3 agonist. Herein, we describe the syntheses and structure-activity relationships of analogues of 1 as well as their in vivo activity in animal models of epilepsy and neuropathic pain. PMID- 20020711 TI - Time-resolved fluorescence study of aggregation-induced emission enhancement by restriction of intramolecular charge transfer state. AB - Cyano-substituted oligo (alpha-phenylenevinylene)-1,4-bis(R-cyano-4 diphenylaminostyryl)-2,5-diphenylbenzene (CNDPASDB) molecules are studied in solution and aggregate state by time-resolved fluorescence techniques. CNDPASDB exhibits a strong solvent polarity dependent characteristic of aggregation induced emission (AIE). By time-dependent spectra, the gradual transition from local excited state to intramolecular charge transfer state with the increasing solvent polarity is clearly resolved. The transition time in high polarity solvent DMF is very fast, around 0.5 ps, resulting in a low fluorescence quantum yield. While in aggregate state, the intramolecular torsion is restricted and the local environment becomes less polar. Thus, the intramolecular charge transfer state is eliminated and efficient AIE occurs. PMID- 20020712 TI - Control of muscle differentiation by a mitochondria-targeted fluorophore. AB - During muscle differentiation, mitochondria undergo dramatic changes in their morphology and distribution to prepare for the higher rate of energy consumption. By applying a mitochondria-targeted rosamine library in C2C12 myogenesis, we discovered one compound that controls muscle differentiation. When treated to undifferentiated myoblasts, our selected compound, B25, inhibited myotube formation, and when treated to fully differentiated myotubes, it induced fission of multinucleated myotubes into mononucleated fragments. Compared to myoseverin, which is known for inducing myotube fission by destabilizing microtubules, B25 affects neither microtubule stability nor cell cycle. Further investigation identified that B25 induces myotube fission through the activation of NF-kappaB, which is one of the important signaling pathways linked to skeletal muscle differentiation. So far, the use of small-molecule fluorophores is limited in the discovery of labeling agents or sensors. In addition to their potential as a sensor, here we show the application of fluorescent small molecules in the discovery of a bioactive probe that induces a specific cellular response. PMID- 20020713 TI - Oligomers of the prion protein fragment 106-126 are likely assembled from beta hairpins in solution, and methionine oxidation inhibits assembly without altering the peptide's monomeric conformation. AB - A portion of the prion protein, PrP106-126, is highly conserved among various species and is thought to be one of the key domains involving amyloid formation of the protein. We used ion mobility spectrometry-mass spectrometry (IMS-MS) in conjunction with replica exchange molecular dynamics (REMD) to examine the monomeric and oligomeric structures of normal PrP106-126 and two nonaggregating forms of the peptide, an oxidized form in which both methionine residues are oxidized to methionine sulfoxide and a control peptide consisting of the same amino acids as PrP106-126 in a scrambled sequence. Our ion mobility and simulation data indicate the presence of a population of beta-hairpin monomers for the normal and oxidized peptides. This is supported by our CD data indicating that a monomer solution of the normal peptide contains approximately 46% beta sheet and approximately 23% beta-turn content, in excellent agreement with our REMD simulations. Oligomerization was seen by IMS-MS for the normal peptide only, not the oxidized peptide or the control sequence. Both our IMS-MS and CD data suggest that this oligomerization results from the association of ordered beta hairpin monomers rather than disordered monomers. Structural analysis shows that the normal and oxidized peptides have similar secondary and tertiary structural properties, suggesting that the inhibition of aggregation caused by methionine oxidation stems from mediating interpeptide interactions rather than by altering the peptide's monomeric conformation. In contrast, an increase in alpha-helical and random coil structural components relative to the normal peptide might be responsible for the lack of observed aggregation of the control peptide. PMID- 20020714 TI - Drug and drug candidate building block analysis. AB - Drug likeness analysis is widely used in modern drug design. However, most drug likeness filters, represented by Lipinski's "Rule of 5", are based on drugs' simple structural features and some physiochemical properties. In this study, we conducted thorough structural analyses for two drug datasets. The first dataset, ADDS, is composed of 1240 FDA-approved drugs, and the second drug dataset, EDDS, is a nonredundant collection of FDA-approved drugs and experimental drugs in different phases of clinical trials from several drug databases (6932 entries). For each molecule, all possible fragments were enumerated using a brutal force approach. Three kinds of building blocks, namely, the drug scaffold, ring system, and the small fragment, were identified and ranked according to the frequencies of their occurrence in drug molecules. The major finding is that most top fragments are essentially common for both drug datasets; the top 50 fragments cover 52.6% and 48.6% drugs for ADDS and EDDS, respectively. The identified building blocks were further ranked according to their relative hit rates in the drug datasets and in a screening dataset, which is a nonredundant collection of screening compounds from many resources. In comparison with the previous reports in the field, we have identified many more high-quality building blocks. The results obtained in this study could provide useful hints to medicinal chemists in designing drug-like compounds as well as prioritizing screening libraries to filter out those molecules lack of functional building blocks. PMID- 20020715 TI - LoFT: similarity-driven multiobjective focused library design. AB - We present LoFT, a tool for focused combinatorial library design. LoFT provides a set of algorithms, constructing a focused library from a chemical fragment space under optimization of multiple design criteria. A weighted multiobjective scoring function based on physicochemical descriptors is employed for traversing the chemical search space. The new aspect of LoFT is that a similarity-driven product based library design approach is provided on fragment level. For this reason the feature tree descriptor is incorporated for similarity comparison of library compounds to given bioactive molecules as well as for diversifying the resulting libraries. The feature tree descriptor abstracts the molecular graph to a tree structure where the nodes are labeled with physicochemical properties. For comparison, the nodes of two trees are mapped onto each other. This strictly hierarchical mechanism is suitable for the efficient comparison of chemical fragments, allowing the evaluation of the resulting products on fragment level without explicitly enumerating them. LoFT was validated, applying three different data sets. Starting with a random reagent selection, we optimized the libraries using maximum similarity to known bioactive molecules and iteratively adding further criteria. Moreover, we compared these results with data we obtained with FTrees-FS. PMID- 20020716 TI - Kinetically controlled photoinduced electron transfer switching in Cu(I) responsive fluorescent probes. AB - Copper(I)-responsive fluorescent probes based on photoinduced electron transfer (PET) switching consistently display incomplete recovery of emission upon Cu(I) binding compared to the corresponding isolated fluorophores, raising the question of whether Cu(I) might engage in adverse quenching pathways. To address this question, we performed detailed photophysical studies on a series of Cu(I) responsive fluorescent probes that are based on a 16-membered thiazacrown receptor ([16]aneNS(3)) tethered to 1,3,5-triarylpyrazoline-fluorophores. The fluorescence enhancement upon Cu(I) binding, which is mainly governed by changes in the photoinduced electron transfer (PET) driving force between the ligand and fluorophore, was systematically optimized by increasing the electron withdrawing character of the 1-aryl-ring, yielding a maximum 29-fold fluorescence enhancement upon saturation with Cu(I) in methanol and a greater than 500-fold enhancement upon protonation with trifluoroacetic acid. Time-resolved fluorescence decay data for the Cu(I)-saturated probe indicated the presence of three distinct emissive species in methanol. Contrary to the notion that Cu(I) might engage in reductive electron transfer quenching, femtosecond time-resolved pump-probe experiments provided no evidence for formation of a transient Cu(II) species upon photoexcitation. Variable temperature (1)H NMR experiments revealed a dynamic equilibrium between the tetradentate NS(3)-coordinated Cu(I) complex and a ternary complex involving coordination of a solvent molecule, an observation that was further supported by quantum chemical calculations. The combined photophysical, electrochemical, and solution chemistry experiments demonstrate that electron transfer from Cu(I) does not compete with radiative deactivation of the excited fluorophore, and, hence, that the Cu(I)-induced fluorescence switching is kinetically controlled. PMID- 20020718 TI - Photodissociation of cis-, trans-, and 1,1-dichloroethylene in the ultraviolet range: characterization of Cl((2)P(J)) elimination. AB - By using photofragment velocity imaging detection coupled with a (2 + 1) resonance-enhanced multiphoton ionization technique, the elimination channel of spin-orbit chlorine atoms in photodissociation of cis-, trans-, and 1,1 dichloroethylene at two photolysis wavelengths of 214.5 and 235 nm is investigated. Translational energy and angular distributions of Cl((2)P(J)) fragmentation are acquired. The Cl((2)P(J)) fragments are produced by two competing channels. The fast dissociation component with higher translational energy is characterized by a Gaussian distribution, resulting from a curve crossing of the initially excited (pi, pi*) state to nearby repulsive (pi, sigma*) and/or (n, sigma*). In contrast, the slow component with a lower translational energy is characterized by a Boltzmann distribution, which dissociates on the vibrationally hot ground state relaxed from the (pi, pi*) state via internal conversion. cis-C(2)H(2)Cl(2) is found to have a larger branching of Boltzmann component than the other two isomers. The fraction of available energy partitioning into translation increases along the trend of cis- < trans- < 1,1-C(2)H(2)Cl(2). This trend may be fitted by a rigid radical model and interpreted by means of a torque generated during the C-Cl bond cleavage. The anisotropy parameters are determined, and the transition dipole moments are expected to be essentially along the C horizontal lineC bond axis. The results are also predicted theoretically. The relative quantum yields of Cl((2)P(J)) have a similar value for the three isomers at the two photolysis wavelengths. PMID- 20020719 TI - Sample cooling or rotation improves C60 organic depth profiles of multilayered reference samples: results from a VAMAS interlaboratory study. AB - We demonstrate two methods to improve the quality of organic depth profiling by C(60) sputtering using multilayered reference samples as part of a VAMAS (Versailles project on Advanced Materials and Standards) interlaboratory study. Sample cooling was shown previously to be useful in extending the useful depth over which organic materials can be profiled. We reinforce these findings and demonstrate that cooling results in a lower initial sputtering yield to approximately -40 degrees C, but the improvement in useful profiling depth continues as the sample is cooled further, even though there is no further reduction in the initial sputtering yield. We report, for the first time, the use of sample rotation in organic depth profiling and demonstrate that the initial sputtering yield at room temperature is maintained throughout the depth of the samples used in this study. Useful profiling depth and good depth resolution are both associated with a constant sputtering yield. The fact that rotation results in the maintenance of depth resolution underlines the fact that depth resolution is often limited by the development of ion-beam-induced topography. Constant sputtering yield results in a constant secondary-ion yield, after transient processes have occurred, and this allows simple quantification methods to be applied to organic depth profiling data. PMID- 20020720 TI - Adsorption of small molecules with the hydroxyl group on sodium halide cluster ions. AB - We have investigated adsorption of molecules with hydroxyl group, ROH, on sodium halide cluster ions, Na(n)X(n-1)(+) (X = F and I, n = 10-17) by mass spectrometry and by theoretical calculations. From analysis of the cluster ion intensities, the adsorption of one water molecule (R = H) is most efficient for Na(13)X(12)(+), whose structure has a NaX defect from a 3 x 3 x 3 cubic structure of n = 14. This result suggests that the defect has an important role in the adsorption reaction. However, it is also found that the reactivity diminishes with increasing bulk size of the R group from H to CH(3), (CH(3))(2)CH, and (CH(3))(3)C. These results imply that the adsorption reactivity is dominated by steric hindrance; the smaller molecules are adsorbed inside the basket structures of Na(13)X(12)(+). Reactivity dependence on the basket size is also discussed by comparing the results of Na(n)F(n-1)(+) and Na(n)I(n-1)(+). PMID- 20020717 TI - Advances in the biology and chemistry of sialic acids. AB - Sialic acids are a subset of nonulosonic acids, which are nine-carbon alpha-keto aldonic acids. Natural existing sialic acid-containing structures are presented in different sialic acid forms, various sialyl linkages, and on diverse underlying glycans. They play important roles in biological, pathological, and immunological processes. Sialobiology has been a challenging and yet attractive research area. Recent advances in chemical and chemoenzymatic synthesis, as well as large-scale E. coli cell-based production, have provided a large library of sialoside standards and derivatives in amounts sufficient for structure-activity relationship studies. Sialoglycan microarrays provide an efficient platform for quick identification of preferred ligands for sialic acid-binding proteins. Future research on sialic acid will continue to be at the interface of chemistry and biology. Research efforts not only will lead to a better understanding of the biological and pathological importance of sialic acids and their diversity but also could lead to the development of therapeutics. PMID- 20020721 TI - Computational studies of transition metal selectivity of octapeptide repeat region of prion protein (PrP). AB - We have presented a detailed theoretical (density functional theory and ONIOM) study on the structure and M-PrP (prion protein) interaction on various prion models, M(PrP) systems (where PrP = HGG, HGGGW x 3H(2)O, and PrP(61-84), and M = Zn, Cu, Ni, Co, Fe, Mn). It was shown that the geometry of the complex [Mn(HGGGW)(H(2)O)] x 2H(2)O is quite different for M = Mn(III) and Mn(II), and partial unfolding occurs only for the M = Mn(III). In [Zn{PrP(61-84)}], Zn(II) forms a bond with carbonyl oxygen of the Pro(84) residue in the axial position. The coordination of transition metal ions to PrP(61-84) induces the significant geometrical changes in the PrP(61-68) and Prp(69-76) octapeptide repeat regions. The Trp(65) and Trp(73) residues come closer to each other. The glutamines Gln(67) and Gln(75) also come closer to one another and result in the formation of a hydrogen bond between the carbonyl carbon of Gln(67) and one of the NH(2) hydrogens of Gln(75). A specific aggregatory effect is found for Co(II) and Mn(II). The relative binding ability of the metal ions increases in the order Zn(II) < Mn(II) < Cu(II) < Fe(II) < Co(II) < Ni(II). The PrP is most distorted upon binding to Co(II) and least distorted upon binding to Zn(II). PMID- 20020722 TI - Bonelike apatite formation utilizing carbon nanotubes as template. AB - Template-induced hydroxyapatite (HA) has broad prospects in the applied fields of regenerative medicine and bone repair. HA thin coatings have been deposited on vertically aligned multiwalled carbon nanotubes (CNTs) via the high-temperature radio-frequency (rf) magnetron sputtering deposition technique. Simulated body fluid (SBF) solution has been used to soak and incubate the HA/CNTs nanocomposites at 37 degrees C. SEM, EDS, XRD, and FTIR characterizations revealed bonelike apatite formation on top of HA/CNTs composites. Coating HA material on well-aligned CNT-template provides a way of combining the superior mechanical properties and chemical stability of the CNTs with the excellent biochemical properties of HA. PMID- 20020723 TI - Pickering emulsions stabilized by a lipophilic surfactant and hydrophilic platelike particles. AB - Liquid paraffin-water emulsions were prepared by homogenizing oil phases containing sorbitan oleate (Span 80) and aqueous phases containing layered double hydroxide (LDH) particles or Laponite particles. While water-in-oil (w/o) emulsions are obtained by combining LDH with Span 80, the emulsions stabilized by Laponite-Span 80 are always o/w types regardless of the Span 80 concentration. Laser-induced fluorescent confocal micrographs indicate that particles are absorbed on the emulsion surfaces, suggesting all the emulsions are stabilized by the particles. The difference of the particle-stabilized emulsion type may be explained by comparing particle contact angles and the oil-water interfacial tensions, indicating that more Span 80 molecules are adsorbed on the LDH particles than on Laponite. Apparently, the LDH particles are rendered more hydrophobic by Span 80, resulting in the formation of w/o emulsions. The long term stability of the emulsions was also compared. Emulsions stabilized by Span 80 alone completely separate into two bulk phases of oil and water after 3 months. However, emulsion stability is greatly enhanced with the addition of LDH or Laponite particles. This synergism was accounted for by an increase of the dilational viscoelasticity modulus of the oil-water interface after particles were added to the aqueous phase. This increase indicates that the gel-like particle layer stays at the oil-water interface and resists emulsion coalescence. Scanning electron microscope (SEM) images display the presence of a firm layer surrounding the emulsion droplets and a three-dimensional particle network which extends into the bulk phase aiding emulsion stability. PMID- 20020724 TI - Electrochemistry on a localized surface plasmon resonance sensor. AB - The optical signal of a localized surface plasmon resonance (LSPR)-based sensor combined with electrochemistry was investigated. Gold nanoparticles were immobilized on an indium tin oxide (ITO) substrate, which functioned as working electrode. Using cyclic voltammetry synchronized with LSPR sensing, surface reactions on gold were detected both electrically and optically. In the capacitive charging regime, optical signals linear to the applied potential were detected. Gold was found to be dissolved above the oxidation potential and partially redeposited during the reduction, which changed size and conformation of the gold nanoparticles. In kinetic measurements, slower potential establishment was observed at lower salt concentrations. Simulations by multiple multipole program (MMP) suggested the formation of a lossy layer by combination of charge depletion of gold and negative ion adsorption even below the reaction potential. We consider the results presented here of importance for any future sensors based on combined plasmonics and electrochemistry. PMID- 20020725 TI - Interfacial binding dynamics of bee venom phospholipase A2 investigated by dynamic light scattering and quartz crystal microbalance. AB - Bee venom phospholipase A(2) (bvPLA(2)) is part of the secretory phospholipase A(2) (sPLA(2)) family whose members are active in biological processes such as signal transduction and lipid metabolism. While controlling sPLA(2) activity is of pharmaceutical interest, the relationship between their mechanistic actions and physiological functions is not well understood. Therefore, we investigated the interfacial binding process of bvPLA(2) to characterize its biophysical properties and gain insight into how membrane binding affects interfacial activation. Attention was focused on the role of membrane electrostatics in the binding process. Although dynamic light scattering experiments indicated that bvPLA(2) does not lyse lipid vesicles, a novel, nonhydrolytic activity was discovered. We employed a supported lipid bilayer platform on the quartz crystal microbalance with dissipation sensor to characterize this bilayer-disrupting behavior and determined that membrane electrostatics influence this activity. The data suggest that (1) adsorption of bvPLA(2) to model membranes is not primarily driven by electrostatic interactions; (2) lipid desorption can follow bvPLA(2) adsorption, resulting in nonhydrolytic bilayer-disruption; and (3) this desorption is driven by electrostatic interactions. Taken together, these findings provide evidence that interfacial binding of bvPLA(2) is a dynamic process, shedding light on how membrane electrostatics can modulate interfacial activation. PMID- 20020726 TI - Constructing a superhydrophobic surface on polydimethylsiloxane via spin coating and vapor-liquid sol-gel process. AB - In this study, a superhydrophobic surface on polydimethylsiloxane (PDMS) substrate was constructed via the proposed vapor-liquid sol-gel process in conjunction with spin coating of dodecyltrichlorosilane (DTS). Unlike the conventional sol-gel process where the reaction takes place in the liquid phase, layers of silica (SiO(2)) particles were formed through the reaction between the reactant spin-coated on the PDMS surface and vapor of the acid solution. This led to the SiO(2) particles inlaid on the PDMS surface. Followed by subsequent spin coating of DTS solution, the wrinkle-like structure was formed, and the static contact angle of the water droplet on the surface could reach 162 degrees with 2 degrees sliding angle and less than 5 degrees contact angle hysteresis. The effect of layers of SiO(2) particles, concentrations of DTS solution and surface topography on superhydrophobicity of the surface is discussed. PMID- 20020727 TI - Naked-eye detection and quantification of heparin in serum with a cationic polythiophene. AB - A strategy for naked-eye detection and quantification of heparin in biological media, such as fetal bovine serum (FBS) is demonstrated by monitoring the absorbance change of a water-soluble cationic polythiophene. The negatively charged heparin interacts with positively charged polythiophene through electrostatic interaction, which leads to polymer conformation and color change from yellow to orange in solution. Under optimized conditions, addition of heparin derivatives, such as hyaluronic acid or chondroitin 4-sulfate to the same polymer solution leads to less change in polymer conformation and solution color due to their lower charge density as compared to that of heparin. Increasing the detection temperature or simply adding some organic solvent to the aqueous media reduces the polymer-polymer interchain pi stacking, and the polymer color change can be used to clearly differentiate heparin from its analogues in homogeneous solutions. Quantification of heparin is also demonstrated by correlating the changes in polymer absorbance to the heparin concentration. A linear calibration curve is observed in the 0-6.7 U/mL and 0-2.2 U/mL ranges for heparin quantification in pure water and in FBS, respectively. PMID- 20020728 TI - Crystal growth, transport, and the structural and magnetic properties of Ln(4)FeGa(12) with Ln = Y, Tb, Dy, Ho, and Er. AB - Ln(4)FeGa(12), where Ln is Y, Tb, Dy, Ho, and Er, prepared by flux growth, crystallize with the cubic Y(4)PdGa(12) structure with the Im3m space group and with a = 8.5650(4), 8.5610(4), 8.5350(3), 8.5080(3), and 8.4760(3) A, respectively. The crystal structure consists of an iron-gallium octahedra and face-sharing rare-earth cuboctahedra of the Au(3)Cu type. Er(4)Fe(0.67)Ga(12) is iron-deficient, leading to a distortion of the octahedral and cuboctahedral environments due to the splitting of the Ga2 site into Ga2 and Ga3 sites. Further, interstitial octahedral sites that are unoccupied in Ln(4)FeGa(12) (Ln = Y, Tb, Dy, and Ho) are partially occupied by Fe2. Y(4)FeGa(12) exhibits weak itinerant ferromagnetism below 36 K. In contrast, Tb(4)FeGa(12), Dy(4)FeGa(12), Ho(4)FeGa(12), and Er(4)Fe(0.67)Ga(12) order antiferromagnetically with maxima in the molar magnetic susceptibilities at 26, 18.5, 9, and 6 K. All of the compounds exhibit metallic electric resistivity, and their iron-57 Mossbauer spectra, obtained between 4.2 and 295 K, exhibit a single-line absorption with a 4.2 K isomer shift of ca. 0.50 mm/s, a shift that is characteristic of iron in an iron gallium intermetallic compound. A small but significant broadening in the spectral absorption line width is observed for Y(4)FeGa(12) below 40 K and results from the small hyperfine field arising from its spin-polarized itinerant electrons. PMID- 20020729 TI - Solvent properties governing solute partitioning in polymer/polymer aqueous two phase systems: nonionic compounds. AB - The solvatochromic solvent parameters characterizing the solvent polarity (pi*), solvent hydrogen-bond donor acidity (alpha), and solvent hydrogen-bond acceptor basicity (beta) of aqueous media were measured in the coexisting phases of nine different aqueous polymer/polymer two-phase systems (ATPS), containing 0.15 M NaCl in 0.01 M phosphate buffer, pH 7.4. Partitioning coefficients of six neutral compounds were measured in the nine ATPS at particular polymer concentrations. The solvatochromic equation was used to describe the partitioning of each compound. Three descriptors of the solvent properties of the phases could describe adequately the partitioning of the solutes in all the ATPS employed. PMID- 20020730 TI - Pseudotetragonal structure of Li(2+x)Ce(x)(3+)Ce(12-x)(4+)F(50): the first mixed valence cerium fluoride. AB - The crystal structure of the new Li(5.5)Ce(12)F(50) compound has been fully characterized by single-crystal and synchrotron powder X-ray diffraction. An accurate pseudotetragonal structure was described in the monoclinic P2(1) space group with 68 independent crystallographic sites. The Li(5.5)Ce(12)F(50) composition belongs to the Li(2+x)Ce(x)(3+)Ce(12-x)(4+)F(50) solid solution. Its structure consists of an opened fluorine framework where a channel network allows the intercalation of relatively mobile lithium cations, inducing the formation of the mixed-valence cerium (the intercalation of Li(+) leads to the reduction of a part of Ce(4+) to Ce(3+)). One part of the lithium ions, necessary for the electroneutrality of the tetravalent equivalent cerium fluoride (Li(2)Ce(12)F(50) composition), is in a locked fluorine polyhedron. Only the supplementary x amount of lithium is able to be exchanged in Li(2+x)Ce(x)(3+)Ce(12-x)(4+)F(50). The structure of Li(2+x)Ce(x)(3+)Ce(12-x)(4+)F(50) is a rearrangement, due to lithium intercalation, of the base CeF(4) structure. Bond valence calculation on Ce sites, Ce coordination polyhedra volumes, and a calculated Ce cationic radius give the indication of a partial long-range ordering of trivalent and tetravalent cerium cations in specific slabs of the structure. (7)Li NMR spectroscopy and XPS analyses have confirmed all of the structure details. PMID- 20020731 TI - Nanometer-scale water-soluble macrocycles from nanometer-sized amino acids. AB - This paper introduces the unnatural amino acids m-Abc(2K) and o-Abc(2K) as nanometer-sized building blocks for the creation of water-soluble macrocycles with well-defined shapes. m-Abc(2K) and o-Abc(2K) are homologues of the nanometer sized amino acid Abc(2K), which we recently introduced for the synthesis of water soluble molecular rods of precise length (J. Am. Chem. Soc. 2007, 129, 7272). Abc(2K) is linear (180 degrees), m-Abc(2K) creates a 120 degree angle, and o Abc(2K) creates a 60 degree angle. m-Abc(2K) and o-Abc(2K) are derivatives of 3' amino-(1,1'-biphenyl)-4-carboxylic acid and 2'-amino-(1,1'-biphenyl)-4-carboxylic acid, with two propyloxyammonium side chains for water solubility. m-Abc(2K) and o-Abc(2K) are prepared as Fmoc-protected derivatives Fmoc-m-Abc(2K(Boc))-OH (1a) and Fmoc-o-Abc(2K(Boc))-OH (1b). These derivatives can be used alone or in conjunction with Fmoc-Abc(2K(Boc))-OH (1c) as ordinary amino acids in Fmoc-based solid-phase peptide synthesis. Building blocks 1a-c were used to synthesize macrocyclic "triangles" 9a-c, "parallelograms" 10a,b, and hexagonal "rings" 11a d. The macrocycles range from a trimer to a dodecamer, with ring sizes from 24 to 114 atoms, and are 1-4 nm in size. Molecular modeling studies suggest that all the macrocycles except 10b should have well-defined triangle, parallelogram, and ring shapes if all of the amide linkages are trans and the o-alkoxy substituents are intramolecularly hydrogen bonded to the amide NH groups. The macrocycles have good water solubility and are readily characterized by standard analytical techniques, such as RP-HPLC, ESI-MS, and NMR spectroscopy. (1)H and (13)C NMR studies suggest that the macrocycles adopt conformations with all trans-amide linkages in CD(3)OD, that the "triangles" and "parallelograms" maintain these conformations in D(2)O, and that the "rings" collapse to form conformations with cis-amide linkages in D(2)O. PMID- 20020732 TI - Controlling surface energy and wetability with Diels-Alder chemistry. AB - Reversible Diels-Alder chemistry was utilized to manipulate the surface energy of glass substrates. Hydrophobic dieneophiles were prepared and attached to glass slides and capillaries to yield a nonwetting surface. Thermal treatment of the surfaces cleaved the Diels-Alder linkage, and resulted in the fabrication of a hydrophilic surface. Preliminary analysis utilized contact angle (CA) measurements to monitor the change in surface energy, and observed a hydrophilic state (CA - 70 +/- 3 degrees) before attachment of the dieneophile to a hydrophobic state (CA - 101 +/- 9 degrees) followed by regeneration of the hydrophilic state (CA - 70 +/- 6 degrees) upon cleavage of the Diels-Alder linkage. The treatments were then applied to glass capillaries, with effective treatment confirmed by fluid column measurements. Patterned treatments were also demonstrated to provide effective flow gating. Finally, attempts to create self pressurizing capillaries were unsuccessful due to pronounced contact angle hysteresis for the hydrophobic surface treatment. PMID- 20020733 TI - Stereoselective aldol reaction of glutarimides using pseudo C(2) symmetry. AB - The boron aldol reaction of beta-substituted glutaric imides bearing an oxazolidinone-based auxiliary proceeds with excellent diastereoselectivity; switching the tertiary amine employed between i-Pr(2)EtN or Et(3)N affords enantiomeric lactone product. PMID- 20020734 TI - Phase separation of lipid microdomains controlled by polymerized lipid bilayer matrices. AB - We developed a micropatterned model biological membrane on a solid substrate that can induce phase separation of lipid microdomains in a designed geometry. Micropatterned lipid bilayers were generated by the photolithographic polymerization of a diacetylene phospholipid, 1,2-bis(10,12-tricosadiynoyl)-sn glycero-3-phosphocholine (DiynePC). By changing the UV dose for the photopolymerization, we could modulate the coverage of the surface by the polymeric bilayer domains. After removing nonpolymerized DiynePC, natural phospholipid membranes were incorporated into the micropatterned polymeric bilayer matrix by a self-assembly process (vesicle fusion). As we incorporated a ternary lipid mixture of 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC), sphingomyelin (SM), and cholesterol (Chol) (1:1:1), liquid ordered domains (Lo: rich in SM and Chol) were accumulated in the polymer free regions, whereas liquid disordered domains (Ld: rich in DOPC) preferentially participated into the partially polymeric bilayer regions. It was postulated that Ld domains preferentially came in contact with the polymeric bilayer boundaries because of their lower elastic moduli and a smaller thickness mismatch at the boundary. The effect of polymeric bilayer matrix to hinder the size growth of Lo domains should also be playing an important role. The controlled phase separation should open new possibilities to locally concentrate membrane proteins and other nanometer sized materials on the substrate by associating them with the lipid microdomains. PMID- 20020735 TI - Total synthesis of pteridic acid A. AB - A convergent approach to the total synthesis of pteridic acid A, having potent plant growth regulator activity, is described. Key steps include the desymmetrization of bicyclic olefin with Brown's chiral hydroboration, acid mediated spiroketalization, and zirconium-catalyzed ethylmagnesation protocol to install the ethyl stereocenter at C14. PMID- 20020736 TI - Biaryl construction through Kumada coupling with diaryl sulfates as one-by-one electrophiles under mild conditions. AB - Diaryl sulfates were successfully applied as one-by-one organo electrophiles in Kumada coupling to construct biaryls with the emission of harmless inorganic salts. PMID- 20020737 TI - Structures and electronic properties of beryllium atom encapsulated in Si(n)((0, 1)) (n = 2-10) clusters. AB - The equilibrium geometries and energies of neutral BeSi(n) (n = 2-10) species and their anions have been studied at the highest level of Gaussian-3 (G3) theory. The results reveal that the ground-state structures of these clusters are Be encapsulated in silicon cages with n >or= 8. The reliable adiabatic electron affinities of BeSi(n) have been predicted to be 1.68 eV for BeSi(2), 1.87 eV for BeSi(3), 2.33 eV for BeSi(4), 2.29 eV for BeSi(5), 2.11 eV for BeSi(6), 2.37 eV for BeSi(7), 2.95 eV for BeSi(8), 2.74 eV for BeSi(9), and 1.92 eV for BeSi(10). The dissociation energies of Be atom from BeSi(n), Si atom from BeSi(n), and Si atom from Si(n) clusters have also been calculated, respectively, to examine relative stabilities. The trend of stability of BeSi(n) changed with n is converse to that of Si(n) when n or= 8, the encapsulated Be atom in silicon cages not only results in an identical trend for stability of BeSi(n) and Si(n) but also improves the stability of Si(n) clusters. PMID- 20020738 TI - Theoretical investigation of the gas-phase reactions of CrO(+) with ethylene. AB - The potential energy surfaces associated with the reactions of chromium oxide cation (CrO(+)) with ethylene have been characterized using density functional, coupled-cluster, and multireference methods. Our calculations show that the most probable reaction involves the formation of acetaldehyde and Cr(+) via a hydride transfer involving the metal center. Our calculations support previous experimental hypotheses that a four-membered ring intermediate plays an important role in the reactivity of the system. We have also characterized a number of viable reaction pathways that lead to other products, including ethylene oxide. Due to the experimental observation that CrO(+) can activate carbon-carbon bonds, a reaction pathway involving C-C bond cleavage has also been characterized. Since many of the reactions involve a change in the spin state in going from reactants to products, locations of these spin surface crossings are presented and discussed. The applicability of methods based on Hartree-Fock orbitals is also discussed. PMID- 20020739 TI - Enhanced laminin-derived peptide AG73-mediated liposomal gene transfer by bubble liposomes and ultrasound. AB - A promising strategy as a cancer therapeutic is tumor-targeted gene delivery. The AG73 peptide derived from the laminin alpha1 chain is a ligand for syndecans, and syndecan-2 is highly expressed in some cancer cells. In this study, AG73-PEG liposomes were developed for selective gene delivery to syndecan-2 overexpressing cancer cells. AG73-PEG liposomes were used in combination with Bubble liposomes and ultrasound exposure to enhance transfection efficiency by promoting the escape of the liposomes from the endosome to the cytosol. AG73-PEG liposomes showed selective gene delivery to syndecan-2 overexpressing cancer cells. Furthermore, AG73-mediated liposomal gene transfection efficiency was enhanced by 60-fold when Bubble liposomes and ultrasound exposure were used, despite the absence of an increase in the uptake of AG73-PEG liposomes into the cells. Confocal microscope analysis revealed that the Bubble liposomes and ultrasound promoted intracellular trafficking of the AG73-PEG liposomes during gene transfection. Thus, the combination of AG73-PEG liposomes with Bubble liposomes and ultrasound exposure may be a promising method to achieve selective and efficient gene delivery for cancer therapy. PMID- 20020740 TI - Transport of ipratropium, an anti-chronic obstructive pulmonary disease drug, is mediated by organic cation/carnitine transporters in human bronchial epithelial cells: implications for carrier-mediated pulmonary absorption. AB - Ipratropium bromide, an anticholinergic drug used for the treatment of asthma and chronic obstructive pulmonary disease, has low oral bioavailability, but systemic exposure, superior to oral administration, can be achieved by inhalation. Therefore, we investigated the pulmonary absorption mechanism of ipratropium using human bronchial epithelial BEAS-2B cells. [3H]Ipratropium uptake by BEAS-2B cells was temperature-dependent and saturable, with a K(m) value of 78.0 microM, suggesting involvement of carrier-mediated uptake. An RT-PCR study showed that organic cation/carnitine transporters OCTN1 and OCTN2 are expressed in BEAS-2B cells, but organic cation transporters (OCTs) are not. Uptake of [3H]ipratropium by HEK293 cells expressing OCTN1 (HEK293/OCTN1) and OCTN2 (HEK293/OCTN2) was significantly increased, compared with mock-transfected cells, and the estimated K(m) values were 444 microM and 53.0 microM, respectively. Finally, the contributions of OCTN1 and OCTN2 to ipratropium uptake were evaluated by measuring [3H]ipratropium uptake by BEAS-2B cells in which OCTN1 or OCTN2 gene expression had been silenced. Knock-down of OCTN1 or OCTN2 suppressed the uptake of [3H]ipratropium to 78.2% and 14.8% of that by control BEAS-2B cells, respectively. In addition, another anticholinergic, tiotropium, was also taken up by both HEK293/OCTN1 and HEK293/OCTN2 cells. Therefore, ipratropium and tiotropium are taken up primarily by OCTN2, and to a lesser extent by OCTN1, in bronchial epithelial cells. These findings are consistent with the pharmacological activity of the drugs after administration via inhalation. PMID- 20020741 TI - Thermodynamic metrics for aggregation of natural resources in life cycle analysis: insight via application to some transportation fuels. AB - While methods for aggregating emissions are widely used and standardized in life cycle assessment (LCA), there is little agreement about methods for aggregating natural resources for obtaining interpretable metrics. Thermodynamic methods have been suggested including energy, exergy, and emergy analyses. This work provides insight into the nature of thermodynamic aggregation, including assumptions about substitutability between resources and loss of detailed information about the data being combined. Methods considered include calorific value or energy, industrial cumulative exergy consumption (ICEC) and its variations, and ecological cumulative exergy consumption (ECEC) or emergy. A hierarchy of metrics is proposed that spans the range from detailed data to aggregate metrics. At the fine scale, detailed data can help identify resources to whose depletion the selected product is most vulnerable. At the coarse scale, new insight is provided about thermodynamic aggregation methods. Among these, energy analysis is appropriate only for products that rely primarily on fossil fuels, and it cannot provide a useful indication of renewability. Exergy-based methods can provide results similar to energy analysis by including only nonrenewable fuels but can also account for materials use and provide a renewability index. However, ICEC and its variations do not address substitutability between resources, causing its results to be dominated by dilute and low-quality resources such as sunlight. The use of monetary values to account for substitutability does not consider many ecological resources and may not be appropriate for the analysis of emerging products. ECEC or emergy explicitly considers substitutability and resource quality and provides more intuitive results but is plagued by data gaps and uncertainties. This insight is illustrated via application to the life cycles of gasoline, diesel, corn ethanol, and soybean biodiesel. Here, aggregate metrics reveal the dilemma facing the choice of fuels: high return on investment versus high renewability. PMID- 20020742 TI - Reactive intermediates formed during olefin polymerization by methylalumoxane activated ansa-zirconocene catalysts: identification of a chain-carrying intermediate by NMR methods. AB - Addition of 1-hexene to methylalumoxane-activated catalyst systems based on rac Me(2)Si(ind)(2)ZrMe(2) causes, concurrent with polyhexene formation and diminution of the otherwise prevalent cation [rac-Me(2)Si(ind)(2)Zr(mu Me)(2)AlMe(2)](+), formation of a hitherto unobserved species [rac Me(2)Si(ind)(2)Zr(mu-R)(mu-Me)AlMe(2)](+), where R is a Zr-bound polyhexyl chain. As hexene is increasingly consumed, this cation decays, mainly back to [rac Me(2)Si(ind)(2)Zr(mu-Me)(2)AlMe(2)](+) and, in part, to some species containing a Zr-bound allylic chain end. PMID- 20020743 TI - Cesium promotion in styrene epoxidation on silver catalysts. AB - The adsorption of a small amount of cesium on Ag(110) redirects the partial oxidation products of styrene from phenylacetaldehyde and phenylketene to styrene oxide. The cesium stabilizes the oxametallacycle intermediate and hinders its transformation to the intermediate that leads to the other products. Cesium does not appear to create any electronic effects on the bonding of the intermediates. Low coverages of cesium induce a (1 x 2) missing-row reconstruction of the entire clean Ag(110) surface and a (3 x 5) surface oxide structure on the cesium reconstructed Ag(110) surface. This (3 x 5)-ordered surface oxide is superimposed on the Ag(111) microfacets produced by the cesium-induced reconstruction, which leads to selectivity and reactivity very similar to those of the extended (111) surface. These studies provide insight into the microscopic origins of the structural effects of cesium in styrene epoxidation on silver catalysts. PMID- 20020744 TI - Verification of biomarkers for diabetic retinopathy by multiple reaction monitoring. AB - Multiple reaction monitoring was used to verify target proteins in 3 groups of vitreous and plasma samples from 3 stages of diabetic retinopathy: macular hole (nondiabetic control), nonproliferative diabetic retinopathy, and proliferative diabetic retinopathy. Twelve target proteins were quantified using triple quadrupole LC-MS/MS and 3 methods to determine the transitions (information dependent analysis, the MIDAS workflow, and the PeptideAtlas database). This study might be the first MRM experiment to analyze large numbers of clinical vitreous and plasma samples for biomarker verification. Consequently, several biomarker candidates were identified for use in further applications. PMID- 20020745 TI - Impact of climate change on three-dimensional dynamic critical load functions. AB - Changes in climate and atmospheric deposition of base cations can alter the ionic composition of soil and surface waters, and therefore affect the structure and function of sensitive ecosystems. However, these drivers are not generally explicitly considered in the calculation of critical loads or dynamic critical loads to evaluate the recovery of ecosystems from elevated acidic deposition. Here we explore the importance of accounting for these changes in calculating dynamic critical loads for ecosystems. We developed three-dimensional dynamic critical load surfaces as a function of nitrate, sulfur, and base cation deposition under current and future climate change scenarios for the Hubbard Brook Experimental Forest, New Hampshire. This case study indicates that dynamic critical loads for nitrate and sulfur will be lower under conditions of potential climate change or decreases in base cation deposition. This analysis suggests that greater emission controls may be needed to protect sensitive forest ecosystems from acidic deposition under a future climate change or conditions of lower atmospheric deposition of base cations, particularly for watersheds experiencing elevated leaching losses of nitrate. This study should facilitate more informed policy decisions on emission control strategies and assessments of ecosystem recovery. PMID- 20020747 TI - Influence of the polymerization potential on the transport properties of polypyrrole films. AB - Polypyrrole films have been prepared by potentiostatic electrochemical polymerization at low temperatures. The cyclic voltammograms and the electronic transport properties of the films are investigated as a function of the polymerization potential. As the potential increases from 520 mV to 1.2 V, the oxidation peak moves to larger voltages, while above 1.2 V, the peak voltage drops again. The film conductivity drops monotonously as the polymerization potential is increased. However, the localization length of the current-carrying states, which characterizes the temperature dependence of the conductivity, correlates with the oxidation peak and shows a minimum for films polymerized at 1.2 V. Furthermore, we show that, with an independent doping step after polymerization, the conductivity of the films can be increased by up to 50%. A maximum conductivity of 1360 S/cm has been observed. PMID- 20020746 TI - Regional electroporation of single cardiac myocytes in a focused electric field. AB - There is now a significant interest in being able to locate single cells within geometrically defined regions of a microfluidic chip and to gain intracellular access through the local electroporation of the cell membrane. This paper describes the microfabrication of electroporation devices which can enable the regional electroporation of adult ventricular myocytes, in order to lower the local electrical resistance of the cell membrane. Initially three different devices, designed to suit the characteristic geometry of the cardiomyocyte, were investigated (all three designs serve to focus the electric field to selected regions of the cell). We demonstrate that one of these three devices revealed the sequence of cellular responses to field strengths of increasing magnitudes, namely, cell contraction, hypercontraction, and lysis. This same device required a reduced threshold voltage for each of these events, including in particular membrane breakdown. We were not only able to show the gradual regional increase in the electric conductivity of the cell membrane but were also able to avoid changes in the local intra- and extracellular pH (by preventing the local generation of protons at the electrode surface, as a consequence of the reduced threshold voltage). The paper provides evidence for new strategies for achieving robust and reproducible regional electroporation, a technique which, in future, may be used for the insertion of large molecular weight molecules (including genes) as well as for on-chip voltage clamping of the primary adult cardiomyocyte. PMID- 20020748 TI - Spectroscopy and photophysics of structural isomers of naphthalene: Z phenylvinylacetylene. AB - The fluorescence spectroscopy of Z-phenylvinylacetylene (Z-PVA) has been studied under jet-cooled conditions. The laser-induced fluorescence (LIF) spectrum shows vibronic activity up to 600 cm(-1) above the pi pi* electronic origin at 33 838 cm(-1). In contrast, the single vibronic level fluorescence spectrum of the electronic origin shows strong intensity in transitions ending in ground state levels at least 1200 cm(-1) above the ground state zero-point level. The double resonance technique of ultraviolet depletion (UVD) spectroscopy was used to show that there are strong absorptions in Z-PVA that are not observed in the LIF spectrum due to the turn of a nonradiative process in this electronic state. The LIF and UVD spectra were compared quantitatively to calculate the relative single vibronic level fluorescence quantum yields. Upon inspection, there are some indications of state specific effects; however, the nature of these effects is unclear. Ab initio and density functional theory calculations of the ground and excited states were used to map the first two excited states of Z-PVA along the C[triple bond]CH bending coordinate, determining them to be pi pi* and pi sigma*, respectively, in character. The crossing of these two states is postulated to be the underlying reason for the observed loss in fluorescence intensity 600 cm(-1) above the pi pi* origin. The spectroscopy of Z-PVA has been compared to the previously characterized E isomer of phenylvinylacetylene [Liu, C. P., Newby, J. J., Muller, C. W., Lee, H. D. and Zwier, T. S. J. Phys. Chem. A 2008, 112 (39), 9454.]. PMID- 20020749 TI - Examining co-based nanocrystals on graphene using low-voltage aberration corrected transmission electron microscopy. AB - We present a method to produce graphene and few layer graphene sheets using solution phase chemistry, which are used as ultrathin support membranes for enhanced imaging of nanomaterials using transmission electron microscopy. We demonstrate this by decorating the surface of the graphene sheets with Co-based nanocrystals (CoCl(2) and hcp Co). Low-voltage aberration-corrected high resolution transmission electron microscopy at 80 kV is used to image the nanocrystals on the thin graphene supports. We show that electron beam irradiation causes the CoCl(2) nanocrystals to become mobile on the graphene surface and exhibit both rotational and translational motion. We provide real time in situ monitoring with atomic resolution of the coalescence of two CoCl(2) nanocrystals on the graphene surface, driven by electron beam irradiation. The CoCl(2) nanocrystals are then annealed in vacuum and transform into Co nanocrystals with hcp crystal structure. We show that these Co nanocrystals are catalytic and electron beam irradiation leads to the etching of the graphene surface, not observed for the CoCl(2) nanocrystals. PMID- 20020750 TI - New urinary metabolites formed from ring-oxidized metabolic intermediates of styrene. AB - The urine from mice exposed to styrene vapors (600 and 1200 mg/m(3), 6 h) was analyzed for ring-oxidized metabolites of styrene. To facilitate the identification of metabolites in urine, the following potential metabolites were prepared: 2-, 3-, and 4-vinylphenol (2-, 3-, and 4-VP), 4-vinylpyrocatechol, and 2-, 3-, and 4-vinylphenylmercapturic acid (2-, 3-, and 4-VPMA). For the analysis of vinylphenols beta-glucuronidase-treated urine was extracted and derivatized with acetanhydride/triethylamine before injection into GC/MS. Three isomers, 2-, 3-, and 4-VP, were found in the exposed urine using authentic standards. Additionally, three novel minor urinary metabolites, arylmercapturic acids 2-, 3 , and 4-VPMA, were identified by LC-ESI-MS(2) by comparison with authentic standards. Excretion of the most abundant isomer, 4-VPMA, amounted to 535 +/- 47 nmol/kg and 984 +/- 78 nmol/kg, representing approximately 0.047 and 0.043% of the absorbed dose for the exposure levels of 600 and 1200 mg/m(3), respectively. The ratio of 2-VPMA, 3-VPMA, and 4-VPMA was approximately 2:1:6. In model reactions of styrene 3,4-oxide (3,4-STO) with N-acetylcysteine in aqueous solutions and of its methyl ester in methanol, 4-vinylphenol was always the main product, while 3-vinylphenol has never been detected. No mercapturic acid was found in the reaction of 3,4-STO with N-acetylcysteine in aqueous solution at pH 7.4 or 9.7, but a small amount of 4-VPMA methyl ester was detected by LC-ESI-MS after the reaction of 3,4-STO with N-acetylcysteine methyl ester. In contrast, no mercapturic acid was found in the reaction of 3,4-STO with N-acetylcysteine in aqueous solution at pH 7.4 or 9.7. These findings indicate a capability of 3,4 STO to react with cellular thiol groups despite its rapid isomerization to vinylphenol in an aqueous environment. Moreover, the in vivo formation of 2- and 3-isomers of both VP and VPMA, neither of which was formed from 3,4-STO in vitro, strongly suggests that another arene oxide, styrene 2,3-oxide, might be a minor metabolic intermediate of styrene. PMID- 20020751 TI - Fabrication and luminescence of designer surface patterns with beta-cyclodextrin functionalized quantum dots via multivalent supramolecular coupling. AB - Supramolecular microcontact printing was used to obtain controlled patterns consisting of quantum dots (QDs) functionalized at their periphery with beta cyclodextrin (beta-CD) in combination with adamantyl terminated dendrimeric "glues". Functionalization of core--shell CdSe/ZnS QDs was achieved by surface ligation. Immobilization of the QDs from solution onto glass substrates printed with (a) adamantyl-terminated poly(propylene imine) dendrimers and (b) via direct microcontact printing of QDs onto the dendrimer layer both yielded stable and robust multilayer structures. The stability of the patterns was primarily due to multivalent supramolecular host--guest interactions between beta-CD located at the QD surface and adamantyl groups at the dendrimer periphery as the dendrimers acted as a "supramolecular glue". The surface-immobilized QDs were capable of forming host--guest complexes with other molecules of interest at binding cavities not occupied by adamantyl groups. Complex formation with ferrocene functionalized molecules at these sites led to partial quenching of the luminescence emission of QDs demonstrating the principle for sensing using the QD multilayer structures. PMID- 20020752 TI - Encapsulating (111)In in nanocontainers for scintigraphic imaging: synthesis, characterization, and in vivo biodistribution. AB - A new strategy for the radiolabeling of porous nanocontainers has been developed, and the first experiments in vivo are reported. Our approach consists of the use of nanometer-sized zeolites whose channels have been filled with the positively charged gamma-emitter (111)In(3+) via simple ion exchange. To avoid leaching of the isotope under physiological conditions, the entrances of the channels have been closed using a specifically designed molecular stopcock. This stopcock has a positively charged group that enters the channels and entraps the loaded radionuclides via electrostatic and steric repulsion. The other side of the stopcock is a bulky triethoxysilane group that can covalently bind to the walls of the zeolite entrances, thereby irreversibly closing the channels. The surface of the zeolites has been functionalized with different chemical groups in order to investigate the different biodistributions depending of the nature of the functionalizations. Preliminary in vivo experiments with Wistar rats have been performed and showed the potential of the approach. This strategy leads to a nanoimaging probe with a very high density of radioisotopes in a confined space, which is highly stable in physiological solution and could allow a large variety of functionalities on its external surface. PMID- 20020753 TI - Interaction of gold nanoparticles with common human blood proteins. AB - In order to better understand the physical basis of the biological activity of nanoparticles (NPs) in nanomedicine applications and under conditions of environmental exposure, we performed an array of photophysical measurements to quantify the interaction of model gold NPs having a wide range of NP diameters with common blood proteins. In particular, absorbance, fluorescence quenching, circular dichroism, dynamic light scattering, and electron microscopy measurements were performed on surface-functionalized water-soluble gold NPs having a diameter range from 5 to 100 nm in the presence of common human blood proteins: albumin, fibrinogen, gamma-globulin, histone, and insulin. We find that the gold NPs strongly associate with these essential blood proteins where the binding constant, K, as well as the degree of cooperativity of particle--protein binding (Hill constant, n), depends on particle size and the native protein structure. We also find tentative evidence that the model proteins undergo conformational change upon association with the NPs and that the thickness of the adsorbed protein layer (bare NP diameter <50 nm) progressively increases with NP size, effects that have potential general importance for understanding NP aggregation in biological media and the interaction of NP with biological materials broadly. PMID- 20020754 TI - Molecular capture in protein nanotubes. AB - We describe molecular capturing properties of protein nanotubes with a controllable ligand binding affinity and size selectivity. These practical biocylinders were prepared using an alternating layer-by-layer (LbL) assembly of protein and oppositely charged poly(amino acid) into the nanoporous polycarbonate (PC) membrane (pore diameter, 400 nm), with subsequent dissolution of the template. The tube wall typically comprises six layers of poly-L-arginine (PLA) and human serum albumin (HSA) [(PLA/HSA)(3)]. Use of high molecular weight PLA (M(w) = ca. 70 000) yielded robust nanotubes, which are available as lyophilized powder. The (PLA/HSA)(3) nanotubes swelled considerably in water, although the outer diameter was almost unaltered. Uranyl ion, 3,3'-diethylthiacarbocyanine iodide, and zinc(II) protoporphyrin IX (ZnPP) were bound to the HSA component in the cylinder wall. Similar nanotubes comprising recombinant HSA mutant [rHSA(His)], which has a strong binding affinity for ZnPP, captured this ligand more tightly. Furthermore, addition of excess myristic acid released ZnPP from the tubes through a ligand replacement reaction. The hybrid nanotubes bearing a single avidin layer as an internal surface captured FITC-biotin efficiently. Biotin-labeled nanoparticles are also incorporated into the tubes when their particle size is sufficiently small to enter the pores. Subsequent TEM observation revealed a line of loaded nanoparticles (100 nm) in the one dimensional space interior. PMID- 20020755 TI - Molassamide, a depsipeptide serine protease inhibitor from the marine cyanobacterium Dichothrix utahensis. AB - A new dolastatin 13 analogue, molassamide (1), was isolated from cyanobacterial assemblages of Dichothrix utahensis collected from the Molasses Reef, Key Largo, Florida, and from Brewer's Bay, St. Thomas, U.S. Virgin Islands. This is the first peptide reported from the cyanobacterial genus Dichothrix and the first natural product isolated from marine Dichothrix spp. Its planar structure was determined by NMR spectroscopic techniques, and the configurations of the asymmetric centers were assigned after chiral HPLC analysis of the hydrolysis products. The depsipeptide 1 exhibited protease-inhibitory activity, with IC(50) values of 0.032 and 0.234 muM against elastase and chymotrypsin, respectively. There was no apparent inhibition of trypsin at 10 microM, the highest concentration tested. PMID- 20020756 TI - RNA-poly(o-methoxyaniline) hybrid templated growth of silver nanoparticles and nanojacketing: physical and electronic properties. AB - Three nanobiocomposites (PRAg31, PRAg11, and PRAg13; the numbers indicate the weight ratios of poly(o-methoxyaniline) (POMA) and ribonucleic acid (RNA), respectively), produced from the same amount of POMA (P) and silver nitrate (AgNO(3)) with differing proportions of RNA (R) are prepared by aging the aqueous solutions of the mixture for 3 weeks at 30 degrees C. The scanning and transmission electron microscopy (SEM and TEM) indicate Ag nanoparticle formation on the hybrid fiber surface and in the PRAg31 system the hybrid fibrils become coated with metallic Ag, the phenomenon being termed as "nanojacketting". The circular dichroism (CD) spectra indicate a small distortion of RNA conformation from A helix toward B helix. FTIR and UV-vis spectra suggest that POMA (emeraldine base, EB) being doped by Ag(+) become oxidized to its pernigraniline base (PB) form and reduce Ag(+) to metallic Ag. The Ag nanoparticles thus produced become stabilized on the fibril surface by co-ordination through nitrogen atoms of POMA (PB) chains. The much slower red shift of pi band to polaron band transition peak in PRAg31 than that of the other two nanobiocomposites is indicative of difficulty in conformational transitions of POMA chain in the "nanojacketted" hybrid fibrils. The dc-conductivity values of the nanobiocomposites are two orders higher than that of the pure POMA-RNA hybrids. The PRAg31 system exhibits rectification property in the I-V characteristic curves and a probable explanation based on the feasibility of p-n junction formation arising from the transfer of lone pair of electrons of nitrogen of POMA (p-type) to the Ag nanoparticles (n-type) has been offered. PMID- 20020757 TI - Enhancement of friction between carbon nanotubes: an efficient strategy to strengthen fibers. AB - Interfacial friction plays a crucial role in the mechanical properties of carbon nanotube based fibers, composites, and devices. Here we use molecular dynamics simulation to investigate the pressure effect on the friction within carbon nanotube bundles. It reveals that the intertube frictional force can be increased by a factor of 1.5-4, depending on tube chirality and radius, when all tubes collapse above a critical pressure and when the bundle remains collapsed with unloading down to atmospheric pressure. Furthermore, the overall cross-sectional area also decreases significantly for the collapsed structure, making the bundle stronger. Our study suggests a new and efficient way to reinforce nanotube fibers, possibly stronger than carbon fibers, for usage at ambient conditions. PMID- 20020758 TI - Aggregation of organic dyes on TiO2 in dye-sensitized solar cells models: an ab initio investigation. AB - A density functional theory (DFT), time-dependent DFT, and ab initio second order Moller-Plesset perturbation theory study of the aggregation of the metal free indoline D102 and D149 dyes on extended TiO(2) models is reported. By selecting the relevant dimeric arrangements on the TiO(2) surface and evaluating, at the same time, the associated spectroscopic response, an almost quantitative description of the extremely different aggregation behavior of the two dyes is provided. Nicely reproducing the experimental evidence, the present results predict strong aggregation interactions and a sizable red-shift of the absorption band in the case of D102, while negligible effects for D149. Our results open the possibility of computationally screening the various aggregation patterns and predicting the corresponding optical response, thus paving the way to an effective molecular engineering of further enhanced sensitizers for solar cell applications. PMID- 20020760 TI - Pd(II)-catalyzed C-H activation/aryl-aryl coupling of phenol esters. AB - Although nitrogen-containing group-directed cyclopalladation reactions have been well-known, Pd(II) insertion into C-H bonds promoted by coordination of an oxygen only group to the palladium remains rather rare. In the present study, the first cyclopalladation complex formed from a simple phenol ester was characterized by X ray crystallography. A promising protocol for the ortho C-H activation/aryl-aryl coupling of phenol esters that was not sensitive to moisture or air was then established. The utility of the reaction was demonstrated for the synthesis of useful phenol derivatives. PMID- 20020759 TI - On the chalcogenophilicity of mercury: evidence for a strong Hg-Se bond in [Tm(Bu(t))]HgSePh and its relevance to the toxicity of mercury. AB - One of the reasons for the toxic effects of mercury has been attributed to its influence on the biochemical roles of selenium. For this reason, it is important to understand details pertaining to the nature of Hg-Se interactions and this has been achieved by comparison of a series of mercury chalcogenolate complexes that are supported by tris(2-mercapto-1-t-butyl-imidazolyl)hydroborato ligation, namely [Tm(Bu(t))]HgEPh (E = S, Se, Te). In particular, X-ray diffraction studies on [Tm(Bu(t))]HgEPh demonstrate that although the Hg-S bonds involving the [Tm(Bu(t))] ligand are longer than the corresponding Cd-S bonds of [Tm(Bu(t))]CdEPh, the Hg-EPh bonds are actually shorter than the corresponding Cd EPh bonds, an observation which indicates that the apparent covalent radii of the metals in these compounds are dependent on the nature of the bonds. Furthermore, the difference in Hg-EPh and Cd-EPh bond lengths is a function of the chalcogen and increases in the sequence S (0.010 A) < Se (0.035 A) < Te (0.057 A). This trend indicates that the chalcogenophilicity of mercury increases in the sequence S < Se < Te. Thus, while mercury is often described as being thiophilic, it is evident that it actually has a greater selenophilicity, a notion that is supported by the observation of facile selenolate transfer from zinc to mercury upon treatment of [Tm(Bu(t))]HgSCH(2)C(O)N(H)Ph with [Tm(Bu(t))]ZnSePh. The significant selenophilicity of mercury is in accord with the aforementioned proposal that one reason for the toxicity of mercury is associated with it reducing the bioavailability of selenium. PMID- 20020761 TI - Interaction of alkaline metal cations with oxidic surfaces: effect on the morphology of SnO2 nanoparticles. AB - Reaction pathways to SnO(2) nanomaterials through the hydrolysis of hydrated tin tetrachloride precursors were investigated. The products were prepared solvothermally starting from hydrated tin tetrachloride and various (e.g., alkali) hydroxides. The influence of the precursor base on the final morphology of the nanomaterials was studied. X-ray powder diffraction (XRD) data indicated the formation of rutile-type SnO(2). Transmission electron microscopy (TEM) studies revealed different morphologies that were formed with different precursor base cations. Data from molecular dynamics (MD) simulations provide theoretical evidence that the adsorption of the cations of the precursor base to the faces of the growing SnO(2) nanocrystals is crucial for the morphology of the nanostructures. PMID- 20020762 TI - Fabrication of hierarchical CaCO3 mesoporous spheres: particle-mediated self organization induced by biphase interfaces and SAMs. AB - Highly ordered hierarchical calcium carbonate is an important phase involved in calcification by a wide variety of invertebrate organisms, and its formation is of technological interest in the development of functional materials. In this article, porous CaCO(3) hierarchical microspheres with a hedgehoglike appearance have been fabricated on the flexible substrate under mild conditions. There are two points that play important roles in the regular organization of the terminal products: one is the biphase interfaces, which are generated by organic solvent n hexane and an aqueous saturated solution of Ca(OH)(2), and the other is hydroxyl terminated monolayers assembled on the flexible PET (poly(ethylene terephthalate)) substrate. The SEM images show that novel CaCO(3) hierarchical microspheres consist of densely stacked "shuttles" by the oriented self organization of CaCO(3) nanoparticles. The IR and XRD spectra indicate that the as-synthesized products are composed of a calcite phase obtained by an ACC (amorphous calcium carbonate)-to-calcite transformation. In view of the results, a nanoparticle-mediated self-organization process induced by biphase interfaces and SAMs template is proposed for the integration of functional materials and nanodevices. PMID- 20020763 TI - Ultrasound-induced in situ formation of coordination organogels from isobutyric acids and zinc oxide nanoparticles. AB - The discovery of ultrasound-induced in-situ formation of coordination organogels using various isobutyric acids (such as isobutyric acid or 2-methylisobutyric acid or 2-bromoisobutyric acid) and zinc oxide nanoparticles was described. FTIR and XRD results suggest that ultrasound irradiation triggers the quick dissolution of zinc oxide nanoparticles by isobutyric acids, resulting in the in situ formation of zinc isobutyrate complexes that undergoes fast sonocrystallization into gel fibers. FESEM results clearly demonstrate the formation of well-defined networks of fibers with several micrometers in length, but the average diameter of the fiber ranges from 30 to 65 nm, depending upon the nature of the isobutyric acids used. A combination of single-crystal structure analysis and powder XRD result was used to envisage the molecular packing present in the gel state. This is probably a very rare case of ultrasound-induced organogelation where metal oxide NPs are used as the precursor. PMID- 20020764 TI - Microplasma discharge ionization source for ambient mass spectrometry. AB - In this paper, we demonstrate the first use of a microplasma ionization source for ambient mass spectrometry. This device is a robust, easy-to-operate microhollow discharge that enables ambient direct analysis of gaseous, liquid, and solid-phase samples with minimum requirements in terms of operating power and high purity gas consumption. The initial performance of the microplasma device has been evaluated by ionizing samples containing dimethyl sulfoxide (DMSO), dimethylformamide (DMF), methyl salicylate, caffeine, l-leucine, l-histidine, loratadine, ibuprofen, acetaminophen, acetylsalicylic acid, and cocaine in various forms. These molecules are diverse in nature, but almost all have relatively high proton affinities. Thus, the major species observed in all obtained mass spectra corresponded to protonated molecules. Though these microplasmas are known to produce significant densities of metastable species and electrons with mean energies greater than several electronvolt, minimal fragmentation was observed. Background spectra showed prominent signals corresponding to H(+)(H(2)O)(2) ions and a distinct lack of H(3)O(+). Small water cluster ions are likely the dominant proton transfer agents, giving rise to mass spectral data very similar to that obtained using other plasma-based ambient ionization techniques. The simplicity, low cost, low power, low rate of gas consumption, and possibility of being batch-fabricated, makes these microplasma devices attractive candidates as ion sources for miniaturized mass spectrometry and other field detection applications. PMID- 20020765 TI - Synthesis and characterization of novel antimicrobial emulsifiers from epsilon polylysine. AB - epsilon-Polylysine (EPL) has been used in the food industry as an antimicrobial additive and also a dietary agent. To generate amphiphilic molecules from EPL, hydrophobically modified epsilon-polylysine graft copolymers, which were denoted as OSA-g-EPLs, were synthesized by reacting EPL with octenyl succinic anhydride (OSA). The success of synthesis was confirmed by (1)H NMR and FT-IR spectroscopy. It was found that OSA-g-EPLs had glass transition temperatures lower than EPL. Furthermore, they were able to form polymer micelles in water and to lower the surface tension of water, confirming their amphiphilic properties. The antimicrobial activities of OSA-g-EPLs were also examined, and the minimum inhibitory concentrations of OSA-g-EPLs against Escherichia coli O157:H7 remained the same as that of EPL. Therefore, OSA-g-EPLs have the potential of becoming bifunctional molecules, which can be used either as surfactants or emulsifiers in the encapsulation of nutraceuticals or drugs or as antimicrobial agents. PMID- 20020766 TI - Cascade nucleophilic addition-cyclic Michael addition of arynes and phenols/anilines bearing ortho alpha,beta-unsaturated groups: facile synthesis of 9-functionalized xanthenes/acridines. AB - A facile synthesis of xanthenes and acridines based on a cascade nucleophilic addition-cyclic Michael addition process of arynes and phenols/anilines substituted with alpha,beta-unsaturated groups at the ortho positions is described. The reaction has also been successfully extended to the synthesis of 9 spiro-xanthene and acridine derivatives with potential biochemical interest. PMID- 20020767 TI - Peptide conjugation at the 5'-end of oligodeoxynucleotides abrogates toll-like receptor 9-mediated immune stimulatory activity. AB - Bacterial and synthetic DNA containing unmethylated CpG motifs act as ligands of Toll-like receptor 9 (TLR9). Our earlier studies showed that 5'-accessibility of synthetic oligodeoxynucleotides containing CpG motif (ODN) is required for TLR9 mediated immune stimulatory activity. Blocking the 5'-end of ODN through conjugation to a variety of moieties reduces immune stimulatory activity (Bioconjugate Chem. 2002, 13, 966-974). In the present study, we conjugated a model peptide, a 28-amino-acid-long beta-amyloid peptide, to either the 5'- or the 3'-end of an ODN via C3 and C6 alkyl linkers. We compared the immune stimulatory activity of the resulting conjugates with that of a parent ODN without conjugation in TLR9-transfected cells, mouse spleen cell cultures, and in vivo in mice. ODN with the peptide conjugated at the 3'-end via C3 and C6 linkers had immune stimulatory activity similar to that of the parent ODN in both in vitro and in vivo in mice. On the contrary, conjugation of peptide at the 5'-end of the ODN significantly abrogated immune stimulatory activity. In conclusion, the results presented here demonstrate that peptide/protein conjugation to ODN is optimal at the 3'-end with either C3 or C6 linker and conjugation at the 5'-end leads to significant loss of TLR9-mediated immune stimulation. PMID- 20020768 TI - Benzannulation from alkyne without metallic catalysts at room temperature to 100 degrees C. AB - An efficient, novel metal-catalyst-free aminobenzannulation of 2-(prop-2 ynyl)(oxo)benzenes 1 with various dialkylamines 2 afforded a variety of 2 aminonaphthalenes 3 in good to excellent yields under mild reaction conditions at room temperature to 100 degrees C (at most). PMID- 20020769 TI - Inhibiting tumor growth by targeting tumor vasculature with galectin-1 antagonist anginex conjugated to the cytotoxic acylfulvene, 6-hydroxylpropylacylfulvene. AB - Targeted delivery of therapeutic drugs promises to become the norm to treat cancer. Here, we conjugated the cytotoxic agent 6-hydroxypropylacylfulvene (HPAF) to anginex, a peptide that targets galectin-1, which is highly expressed in endothelial cells of tumor vessels. In a human ovarian cancer model in mice, the conjugate inhibited tumor growth better than equivalent doses of either compound alone. Immunofluorescence on tumor tissue demonstrated that the conjugate, like parent anginex, selectively targeted tumor vasculature and inhibited tumor angiogenesis. Increased activity from the conjugate further suggests that HPAF retains at least some of its normal cytotoxic activity when linked to anginex. More importantly perhaps is the observation that the conjugate abrogates apparent systemic toxicity from treatment with HPAF. This work contributes to the development of tumor vascular targeting agents against cancer in the clinic. PMID- 20020770 TI - Static magnetic field effect on the Fremy's salt-ascorbic acid chemical reaction studied by continuous-wave electron paramagnetic resonance. AB - Static magnetic field effect in the framework of the radial pair mechanism (RPM) theory was studied on the biologically significant chemical reaction between ascorbic acid and Fremy's salt. The data indicate that the reaction rate depends on the applied magnetic field strength. The time scale of the studied reaction and the improved continuous-wave electron paramagnetic resonance system allowed for the first time the direct comparison of the amplitude differences between exposed and control samples in the strictly same boundary conditions. Until now the RPM was studied in a different time scale, focusing only on faster reactions by time-resolved techniques or by spectrophotometer measurement. The magnetic field effects presently measured can not be extended tout court to living systems; however the understanding of magnetic field sensitivity in basic chemical reaction in vitro could help clarifying the underlying basic step of interaction between magnetic fields and biological systems. PMID- 20020771 TI - Nuclear magnetic resonance parameters of water hexamers. AB - Nuclear magnetic resonance (1)H, (16)O, and (17)O chemical shifts, as well as (17)O quadrupolar parameters in several isomers of water hexamer clusters, are studied using density functional theory calculations and the gauge including projector augmented wave (GIPAW) pseudopotential method. The prism, cage, book, bag, chain, and two cyclic isomers are investigated, and structures with (16)O and (17)O nuclei are examined. It is found that the hydrogen and oxygen chemical shifts show a substantial variation. In six more stable hexamers, all quadrupole coupling constants decrease and asymmetry parameters increase in a comparison with bulk water, whereas a chain isomer shows an opposite behavior. The values of NMR parameters are in reasonable agreement with existing results obtained by more computationally demanding methods. PMID- 20020772 TI - Mass spectrometric analyses of peptides and proteins in human gingival crevicular fluid. AB - Gingival crevicular fluid (GCF) is a pathophysiological fluid that flows into the oral cavity. Human GCF was collected using sterile glass microcapillary tubes from inflamed periodontal sites in patients who had a history of periodontal disease and were in the maintenance phase of treatment. Samples from individual sites were analyzed using MS techniques both before and following HPLC. GCF samples were also pooled and subjected to SDS-PAGE, in-gel digestion and MS analyses using both MALDI-TOF/TOF MS and nanoLC-ESI-MS/MS. MS spectra were used to search human protein sequence databases for protein identification. With these approaches, 33 peptides and 66 proteins were positively identified in human GCF. All of the peptides discovered in this study are reported in GCF here for the first time. Forty-three of the identified proteins, such as actin and the actin binding proteins profilin, cofilin and gelsolin, have not been reported in GCF before. PMID- 20020773 TI - Global analysis of TDP-43 interacting proteins reveals strong association with RNA splicing and translation machinery. AB - TDP-43 is a highly conserved and ubiquitously expressed member of the heterogeneous nuclear ribonucleoprotein (hnRNP) family of proteins. Recently, TDP 43 was shown to be a major disease protein in the ubiquitinated inclusions characteristic of most cases of amyotrophic lateral sclerosis (ALS), tau-negative frontotemporal lobar degeneration (FTLD), and inclusion body myopathy. In these diseases, TDP-43 is redistributed from its predominantly nuclear location to ubiquitin-positive, cytoplasmic foci. The extent to which TDP-43 drives pathophysiology is unknown, but the identification of mutations in TDP-43 in familial forms of ALS and FTLD-U suggests an important role for this protein in pathogenesis. Little is known about TDP-43 function and only a few TDP-43 interacting proteins have been previously identified, which makes further insight into both the normal and pathological functions of TDP-43 difficult. Here we show, via a global proteomic approach, that TDP-43 has extensive interaction with proteins that regulate RNA metabolism. Some interactions with TDP-43 were found to be dependent on RNA-binding, whereas other interactions are RNA-independent. Disease-causing mutations in TDP-43 (A315T and M337V) do not alter its interaction profile. TDP-43 interacting proteins largely cluster into two distinct interaction networks, a nuclear/splicing cluster and a cytoplasmic/translation cluster, strongly suggesting that TDP-43 has multiple roles in RNA metabolism and functions in both the nucleus and the cytoplasm. Finally, we found numerous TDP-43 interactors that are known components of stress granules, and indeed, we find that TDP-43 is also recruited to stress granules. PMID- 20020774 TI - Exploring disease through metabolomics. AB - Metabolomics approaches provide an analysis of changing metabolite levels in biological samples. In the past decade, technical advances have spurred the application of metabolomics in a variety of diverse research areas spanning basic, biomedical, and clinical sciences. In particular, improvements in instrumentation, data analysis software, and the development of metabolite databases have accelerated the measurement and identification of metabolites. Metabolomics approaches have been applied to a number of important problems, which include the discovery of biomarkers as well as mechanistic studies aimed at discovering metabolites or metabolic pathways that regulate cellular and physiological processes. By providing access to a portion of biomolecular space not covered by other profiling approaches (e.g., proteomics and genomics), metabolomics offers unique insights into small molecule regulation and signaling in biology. In the following review, we look at the integration of metabolomics approaches in different areas of basic and biomedical research, and try to point out the areas in which these approaches have enriched our understanding of cellular and physiological biology, especially within the context of pathways linked to disease. PMID- 20020775 TI - Designed proteins to modulate cellular networks. AB - A major challenge of protein design is to create useful new proteins that interact specifically with biological targets in living cells. Such binding modules have many potential applications, including the targeted perturbation of protein networks. As a general approach to create such modules, we designed a library with approximately 10(9) different binding specificities based on a small 3-tetratricopeptide repeat (TPR) motif framework. We employed a novel strategy, based on split GFP reassembly, to screen the library for modules with the desired binding specificity. Using this approach, we identified modules that bind tightly and specifically to Dss1, a small human protein that interacts with the tumor suppressor protein BRCA2. We showed that these modules also bind the yeast homologue of Dss1, Sem1. Furthermore, we demonstrated that these modules inhibit Sem1 activity in yeast. This strategy will be generally applicable to make novel genetically encoded tools for systems/synthetic biology applications. PMID- 20020776 TI - In vivo structure-activity relationship study of dorsomorphin analogues identifies selective VEGF and BMP inhibitors. AB - The therapeutic potential of small molecule signaling inhibitors is often limited by off-target effects. Recently, in a screen for compounds that perturb the zebrafish embryonic dorsoventral axis, we identified dorsomorphin, the first selective inhibitor of bone morphogenetic protein (BMP) signaling. Here we show that dorsomorphin has significant "off-target" effects against the VEGF (vascular endothelial growth factor) type-2 receptor (Flk1/KDR) and disrupts zebrafish angiogenesis. Since both BMP and VEGF signals are known to be involved in vascular development, we sought to determine whether dorsomorphin's antiangiogenic effects are due to its impact on the BMP or VEGF signals through the development of analogues that target BMP but not VEGF signaling and vice versa. In a structure-activity relationship (SAR) study of dorsomorphin analogues based primarily on their effects on live zebrafish embryos, we identified highly selective and potent BMP inhibitors as well as selective VEGF inhibitors. One of the BMP inhibitors, DMH1, which exclusively targets the BMP but not the VEGF pathway, dorsalized the embryonic axis without disrupting the angiogenic process, demonstrating that BMP signaling was not involved in the angiogenic process. This is one of the first full-scale SAR studies performed in vertebrates and demonstrates the potential of zebrafish as an attractive complementary platform for drug development that incorporates an assessment of in vivo bioactivity and selectivity in the context of a living organism. PMID- 20020777 TI - Evaluation of established coronary heart disease on the basis of HDL and non-HDL NMR lipid profiling. AB - A (1)H NMR-based lipid profiling approach was used to investigate the prediction of coronary heart disease (CHD) and examine the confounding effect of factors such as gender, triglycerides, HDL-cholesterol and age levels on the prediction of disease. The HDL and non-HDL lipid profiles in 47 patients with triple vessel disease (TVD) and 41 patients with normal coronary arteries (NCA) both documented angiographically were generated. The presence of CHD was predicted with a sensitivity and specificity of 52% and 75% for HDL model and 78% and 80% for non HDL, respectively. The lipid constituents of HDL lipoproteins which contributed to the separation between the two groups were the saturated fatty acids, cholesterol, total omega-3 fatty acids, degree of unsaturation, diallylic protons from polyunsaturated fatty acids, linoleic acid and, to a lesser extent, the number of fatty acids, triglycerides, unsaturated fatty acids and phosphatidylcholine. Respectively, for non-HDL, lipoproteins were the saturated fatty acids, number of fatty acids, cholesterol, unsaturated fatty acids and phosphatidylcholine. Gender, triglycerides, HDL-cholesterol and age influenced the lipid constituents of HDL and non-HDL lipoproteins that contributed to the separation between subgroups and confounded the predictive power of the models. NMR-based lipid profiling analysis could contribute to the identification of noninvasive markers for the presence and the development of the disease. PMID- 20020778 TI - Proteomic changes in the photoreceptor outer segment upon intense light exposure. AB - Acute light-induced photoreceptor degeneration has been studied in experimental animals as a model for photoreceptor cell loss in human retinal degenerative diseases. Light absorption by rhodopsin in rod photoreceptor outer segments (OS) induces oxidative stress and initiates apoptotic cell death. However, the molecular events that induce oxidative stress and initiate the apoptotic cascade remain poorly understood. To better understand the molecular mechanisms of light induced photoreceptor cell death, we studied the proteomic changes in OS upon intense light exposure by using a proteolytic (18)O labeling method. Of 171 proteins identified, the relative abundance of 98 proteins in light-exposed and unexposed OS was determined. The quantities of 11 proteins were found to differ by more than 2-fold between light-exposed OS and those remaining in darkness. Among the 11 proteins, 8 were phototransduction proteins and 7 of these were altered such that the efficiency of phototransduction would be reduced or quenched during light exposure. In contrast, the amount of OS rhodopsin kinase was reduced by 2-fold after light exposure, suggesting attenuation in the mechanism of quenching phototransduction. Liquid chromatography multiple reaction monitoring (LC-MRM) was performed to confirm this reduction in the quantity of rhodopsin kinase. As revealed by immunofluorescence microscopy, this reduction of rhodopsin kinase is not a result of protein translocation from the outer to the inner segment. Collectively, our findings suggest that the absolute quantity of rhodopsin kinase in rod photoreceptors is reduced upon light stimulation and that this reduction may be a contributing factor to light-induced photoreceptor cell death. This report provides new insights into the proteomic changes in the OS upon intense light exposure and creates a foundation for understanding the mechanisms of light-induced photoreceptor cell death. PMID- 20020779 TI - Assessment of the reproducibility of random hexapeptide peptide library-based protein normalization. AB - The wide dynamic range of proteins in biological samples poses a challenge for the detection of low-abundance proteins. Recently, combinatorial hexapeptide peptide libraries have been suggested as an approach to normalization of proteins in such mixtures. We examined the reproducibility of a commercial hexapeptide ligand library for quantitative and comparative serum proteomic analysis. We also compared this technology with IgY-based affinity depletion. PMID- 20020780 TI - Artificial photosynthesis and solar fuels. PMID- 20020783 TI - Mitochondrial-driven ubiquinone enhances extracellular calcium-dependent nitric oxide production and reduces glycochenodeoxycholic acid-induced cell death in hepatocytes. AB - Ca(2+) mobilization, nitric oxide (NO), and oxidative stress have been involved in cell death induced by hydrophobic bile acid in hepatocytes. The aim of the study was the elucidation of the effect of the antioxidant mitochondrial-driven ubiquinone (Mito Q) on the intracellular Ca(2+) concentration, NO production, and cell death in glycochenodeoxycholic acid (GCDCA)-treated HepG2 cells. The role of the regulation of the intracellular Ca(2+) concentration by Ca(2+) chelators (EGTA or BAPTA-AM), agonist of Ca(2+) entrance (A23187) or NO (L-NAME or NO donor), was assessed during Mito Q cytoprotection in GCDCA-treated HepG2 cells. Cell death, NO synthase (NOS)-1, -2, and -3 expression, Ca(2+) mobilization, and NO production were evaluated. GCDCA reduced the intracellular Ca(2+) concentration and NOS-3 expression and enhanced cell death in HepG2. NO donor prevented and L-NAME enhanced GCDCA-induced cell death. The reduction of Ca(2+) entry by EGTA, but not its release from intracellular stores by BAPTA-AM, reduced the expression of NOS-3 and enhanced cell death in control and GCDCA-treated cells. Mito Q prevented the reduction of intracellular Ca(2+) concentration, NOS 3 expression, NO production, and cell death in GCDCA-treated HepG2 cells. The conclusion is that the recovery of Ca(2+)-dependent NOS-3 expression by Mito Q may be considered an additional cytoprotective property of an antioxidant. PMID- 20020784 TI - Combining hypoxic methods for peak performance. AB - New methods and devices for pursuing performance enhancement through altitude training were developed in Scandinavia and the USA in the early 1990s. At present, several forms of hypoxic training and/or altitude exposure exist: traditional 'live high-train high' (LHTH), contemporary 'live high-train low' (LHTL), intermittent hypoxic exposure during rest (IHE) and intermittent hypoxic exposure during continuous session (IHT). Although substantial differences exist between these methods of hypoxic training and/or exposure, all have the same goal: to induce an improvement in athletic performance at sea level. They are also used for preparation for competition at altitude and/or for the acclimatization of mountaineers. The underlying mechanisms behind the effects of hypoxic training are widely debated. Although the popular view is that altitude training may lead to an increase in haematological capacity, this may not be the main, or the only, factor involved in the improvement of performance. Other central (such as ventilatory, haemodynamic or neural adaptation) or peripheral (such as muscle buffering capacity or economy) factors play an important role. LHTL was shown to be an efficient method. The optimal altitude for living high has been defined as being 2200-2500 m to provide an optimal erythropoietic effect and up to 3100 m for non-haematological parameters. The optimal duration at altitude appears to be 4 weeks for inducing accelerated erythropoiesis whereas <3 weeks (i.e. 18 days) are long enough for beneficial changes in economy, muscle buffering capacity, the hypoxic ventilatory response or Na(+)/K(+)-ATPase activity. One critical point is the daily dose of altitude. A natural altitude of 2500 m for 20-22 h/day (in fact, travelling down to the valley only for training) appears sufficient to increase erythropoiesis and improve sea-level performance. 'Longer is better' as regards haematological changes since additional benefits have been shown as hypoxic exposure increases beyond 16 h/day. The minimum daily dose for stimulating erythropoiesis seems to be 12 h/day. For non-haematological changes, the implementation of a much shorter duration of exposure seems possible. Athletes could take advantage of IHT, which seems more beneficial than IHE in performance enhancement. The intensity of hypoxic exercise might play a role on adaptations at the molecular level in skeletal muscle tissue. There is clear evidence that intense exercise at high altitude stimulates to a greater extent muscle adaptations for both aerobic and anaerobic exercises and limits the decrease in power. So although IHT induces no increase in VO(2max) due to the low 'altitude dose', improvement in athletic performance is likely to happen with high-intensity exercise (i.e. above the ventilatory threshold) due to an increase in mitochondrial efficiency and pH/lactate regulation. We propose a new combination of hypoxic method (which we suggest naming Living High-Training Low and High, interspersed; LHTLHi) combining LHTL (five nights at 3000 m and two nights at sea level) with training at sea level except for a few (2.3 per week) IHT sessions of supra-threshold training. This review also provides a rationale on how to combine the different hypoxic methods and suggests advances in both their implementation and their periodization during the yearly training programme of athletes competing in endurance, glycolytic or intermittent sports. PMID- 20020785 TI - Glycaemic index, glycaemic load and exercise performance. AB - The concept of the glycaemic index (GI) was first introduced in the early 1980s as a method of functionally ranking carbohydrate foods based on their actual postprandial blood glucose response compared with a reference food (either glucose or white bread). Although the GI is a debatable topic among many exercise and health professionals, nutritional recommendations to improve exercise performance and enhance exercise capacity are regularly based on information related to the GI. Studies focusing on the consumption of a pre-exercise GI meal have provided evidence that a benefit exists in relation to endurance performance and substrate utilization when a low GI meal is compared with a high GI meal. However, other investigations have shown that when nutritional strategies incorporating GI are applied to multiple meals, there is no clear advantage to the athlete in terms of exercise performance and capacity. It has been suggested that carbohydrate ingestion during endurance exercise negates the effect of the consumption of pre-exercise GI meals. The glycaemic load (GL) is a relatively novel concept in the area of sports nutrition, and has not been widely investigated. Its premise is that the effect, if any, on exercise performance is determined by the overall glycaemic effect of a diet and not by the amount of carbohydrate alone. The claims for GL have been disputed by a number of sports nutrition specialists, and have gone largely unrecognized by professional and scientific bodies. Research on the effect of the GL on exercise performance and capacity is still at an early stage, but recent studies have shown that the concept may have some merit as far as sports nutrition is concerned. It has been suggested that the GL may be a better predictor of glycaemic responses than the GI alone. PMID- 20020786 TI - The influence of estrogen on skeletal muscle: sex matters. AB - As women enter menopause, the concentration of estrogen and other female hormones declines. This hormonal decrease has been associated with a number of negative outcomes, including a greater incidence of injury as well as a delay in recovery from these injuries. Over the past two decades, our understanding of the protective effects of estrogen against various types of injury and disease states has grown immensely. In skeletal muscle, studies with animals have demonstrated that sex and estrogen may potentially influence muscle contractile properties and attenuate indices of post-exercise muscle damage, including the release of creatine kinase into the bloodstream and activity of the intramuscular lysosomal acid hydrolase, beta-glucuronidase. Furthermore, numerous studies have revealed an estrogen-mediated attenuation of infiltration of inflammatory cells such as neutrophils and macrophages into the skeletal muscles of rats following exercise or injury. Estrogen has also been shown to play a significant role in stimulating muscle repair and regenerative processes, including the activation and proliferation of satellite cells. Although the mechanisms by which estrogen exerts its influence upon indices of skeletal muscle damage, inflammation and repair have not been fully elucidated, it is thought that estrogen may potentially exert its protective effects by: (i) acting as an antioxidant, thus limiting oxidative damage; (ii) acting as a membrane stabilizer by intercalating within membrane phospholipids; and (iii) binding to estrogen receptors, thus governing the regulation of a number of downstream genes and molecular targets. In contrast to animal studies, studies with humans have not as clearly delineated an effect of estrogen on muscle contractile function or on indices of post exercise muscle damage and inflammation. These inconsistencies have been attributed to a number of factors, including age and fitness level of subjects, the type and intensity of exercise protocols, and a focus on sex differences that typically involve factors and hormones in addition to estrogen. In recent years, hormone replacement therapy (HRT) or estrogen combined with exercise have been proposed as potentially therapeutic agents for postmenopausal women, as these agents may potentially limit muscle damage and inflammation and stimulate repair in this population. While the benefits and potential health risks of long-term HRT use have been widely debated, controlled studies using short-term HRT or other estrogen agonists may provide future new and valuable insights into understanding the effects of estrogen on skeletal muscle, and greatly benefit the aging female population. Recent studies with older females have begun to demonstrate their benefits. PMID- 20020787 TI - The pathomechanics, pathophysiology and prevention of cervical spinal cord and brachial plexus injuries in athletics. AB - Cervical spinal cord injuries may occur with catastrophic sequelae (e.g. quadriplegia) in collision sport activities. The discovery was made that the head down tackling technique in football straightens the spine into a position vulnerable for compression and, thus, is responsible for these incidents. This led to rule changes requiring head-up tackling, which in turn resulted in the reduction of the incidence of these injuries. However, the dramatic initial reduction in the occurrence - from 32 and 34 catastrophic injuries in 1975 and 1976, respectively, down to 12 in 1977 - has levelled off with ten and eight reported cases in 2006 and 2007, respectively. The football community has increased their efforts to prevent head-down tackling with additional rule changes. Brachial plexus injury prevention must rely on properly fitted shoulder pads and use of equipment such as 'cowboy' collars. Furthermore, physicians must take into consideration cervical cord neurapraxia, congenital stenosis and other risk factors in patients who wish to return to contact sports. PMID- 20020789 TI - The central governor model cannot be adequately tested by observing its components in isolation. PMID- 20020788 TI - Mountain biking injuries in children and adolescents. AB - Over the last decade, the sport of mountain biking has experienced extensive growth in youth participation. Due to the unpredictable nature of outdoor sport, a lack of rider awareness and increased participation, the number of injuries has unnecessarily increased. Many believe that the actual incidence of trauma in this sport is underestimated and is just the 'tip of the iceberg'. The most common mechanism of injury is usually attributed to downhill riding and forward falling. Although rare, this type of fall can result in serious cranial and thoraco abdominal trauma. Head and neck trauma continue to be documented, often resulting in concussions and the possibility of permanent neurological sequelae. Upper limb injuries range from minor dermal abrasions, contusions and muscular strains to complex particular fracture dislocations. These are caused by attempting to arrest the face with an outstretched hand, leading to additional direct injury. Common overuse injuries include repeated compression from the handlebars and vibration leading to neurovascular complications in the hands. Along with reports of blunt abdominal trauma and lumbar muscle strains, lower extremity injuries may include various hip/pelvic/groin contusions, patellofemoral inflammation, and various muscle strains. The primary causes of mountain biking injuries in children and adolescents include overuse, excessive fatigue, age, level of experience, and inappropriate or improperly adjusted equipment. Additional factors contributing to trauma among this age group involve musculoskeletal immaturity, collisions and falls, excessive speed, environmental conditions, conditioning and fitness status of the rider, nonconservative behavioural patterns, and inadequate medical care. The limited available data restrict the identification and understanding of specific paediatric mountain biking injuries and injury mechanisms. Education about unnecessary risk of injury, use of protective equipment, suitable bikes and proper riding technique, coupled with attentive and proper behaviour, are encouraged to reduce unnecessary injury. This article provides information on the causation and risk factors associated with injury among young mountain bikers, and recommendations to minimize trauma and enhance optimal performance and long-term enjoyment in this outdoor sport. PMID- 20020790 TI - Who should have access to the controlled substances in your office? PMID- 20020791 TI - The effect of local anesthetic on quality of recovery characteristics following dental rehabilitation under general anesthesia in children. AB - This study is a randomized, prospective, double-blind study to evaluate the effects of the combination of local anesthetics and an intravenous nonsteroidal anti-inflammatory drug (NSAID) vs NSAID alone on quality of recovery following dental rehabilitation under general anesthesia (GA). Twenty-seven healthy children aged 3-5.5 years underwent dental rehabilitation under GA. Fifteen children in the experimental group received oral infiltration of local anesthetic in addition to intravenous ketorolac tromethamine, while 12 children in the control group received intravenous ketorolac tromethamine alone for postoperative pain management. Pain behaviors were evaluated immediately postoperatively using a FLACC scale and 4 hours postoperatively by self-report using various scales. Parents reported perception of child pain and comfort and any occurrences of postoperative cheek biting. The use of intraoral infiltration local anesthesia for complete dental rehabilitation under general anesthesia for children aged 3 5.5 years did not result in improved pain behaviors in the postanesthesia care unit (PACU), nor did it result in improved pain behaviors 4-6 hours postoperatively as measured by the FLACC scale, FACES scale, and subjective reports of parents or a PACU nurse. Those children receiving local anesthesia had a higher incidence of negative symptoms related to local anesthetic administration, including a higher incidence of lip and cheek biting, which was of clinical importance, but not statistically significant. Infiltration of local anesthetic for dental rehabilitation under general anesthesia did not improve quality of recovery in children aged 3-5.5 years. PMID- 20020792 TI - Local and systemic toxicity of intraoral submucosal injections of phentolamine mesylate (OraVerse). AB - OraVerse, an injectable formulation of phentolamine mesylate (PM), was recently approved by the U.S. Food and Drug Administration (FDA) for reversal of anesthesia of the lip and tongue and associated functional deficits resulting from an intraoral submucosal injection of a local anesthetic containing a vasoconstrictor. Because PM had not been approved previously for submucosal administration, 2 Good Laboratory Practices (GLP) studies in dogs designed to investigate systemic toxicity and the local effects of single and repeated dosing of OraVerse on the inferior alveolar nerve and branches of the superior alveolar nerve and adjacent soft tissues after local administration were conducted. Systemic toxicity was measured by preinjection and postinjection clinical examinations, clinical chemistry, and gross and microscopic examinations of major organs after necropsy. No evidence of systemic toxicity was detected. Local nerve and adjacent tissue damage was assessed by conventional histopathology. Nerve degeneration was evident in 1 animal. Mild perineural inflammation adjacent to the inferior alveolar nerve and inflammatory exudates were observed in submucosal tissues in several animals. No changes were observed in the nerves at injection sites of dogs from any dose group that were considered directly related to the test articles. These data reveal that single and repeated intraoral administrations of OraVerse are well tolerated in beagle dogs. PMID- 20020793 TI - The efficacy of a repeated buccal infiltration of articaine in prolonging duration of pulpal anesthesia in the mandibular first molar. AB - Previous studies have shown declining rates of pulpal anesthesia over 60 minutes when a cartridge of 4% articaine is used with 1:100,000 epinephrine for buccal infiltration in the mandibular first molar. The authors conducted a prospective, randomized, single-blind, crossover study comparing the degree of pulpal anesthesia obtained with 2 sets of mandibular first molar buccal infiltrations, given in 2 separate appointments, to 86 adult subjects: an initial infiltration of a cartridge of 4% articaine with 1:100,000 epinephrine plus a repeated infiltration of the same anesthetic and dose given 25 minutes following the initial infiltration versus an initial infiltration of a cartridge of 4% articaine with 1:100,000 epinephrine plus a mock repeated infiltration given 25 minutes following the initial infiltration. The authors used an electric pulp tester to test the first molar for anesthesia in 3-minute cycles for 112 minutes after the injections. The repeated infiltration significantly improved pulpal anesthesia from 28 minutes through 109 minutes in the mandibular first molar. A repeated infiltration of a cartridge of 4% articaine with 1:100,000 epinephrine given 25 minutes after an initial infiltration of the same type and dose of anesthetic significantly improved the duration of pulpal anesthesia, when compared with only an initial buccal infiltration, in the mandibular first molar. PMID- 20020794 TI - Preoperative medical evaluation: part 2: pulmonary, endocrine, renal, and miscellaneous considerations. AB - A thorough assessment of a patient's medical status is standard practice when dental care is provided. Although this is true for procedures performed under local anesthesia alone, the information gathered may be viewed somewhat differently if the dentist is planning to provide sedation or general anesthesia as an adjunct to dental treatment. This article, the second of a 2-part sequence on preoperative assessment, will address pulmonary and other noncardiovascular disorders. PMID- 20020797 TI - Meaning and value of productivity to adults with intellectual disabilities. AB - Involvement in productivity roles such as work, volunteerism, and personal projects plays a central role in the lives of most adults and is associated with enhanced physical and mental health. This study examined the meaning of productivity to adults with intellectual disabilities, their satisfaction with the roles they held, and contributors and barriers to achieving satisfying productivity outcomes. The results suggest a number of key areas to address in future research and practice, including systemic and social barriers to choice and meaningful participation. PMID- 20020798 TI - Health profile of aging family caregivers supporting adults with intellectual and developmental disabilities at home. AB - The health status of 206 female caregivers supporting adults with intellectual and developmental disabilities at home was investigated using objective (i.e., presence of chronic health conditions and activity limitations) and subjective (i.e., self-perceived health status) health measures compared with those of women in the general population in 2 age groups: middle age (Ages 40-59 years) and older ages (> or =60 years). Prevalence of arthritis, high blood pressure, obesity, and activity limitations for the caregivers in both age groups was significantly higher than that for women in the general population. Middle-age caregivers reported a higher prevalence of diabetes and high blood cholesterol than their age peers from the general population. Despite the potential health challenges, the caregivers generally perceived their health better than that of women in the general population. Older caregivers' perceptions on their psychological well being, however, appeared to an exception. Implications regarding potential health risks for caregivers and residential services for persons with intellectual and developmental disabilities are discussed. PMID- 20020799 TI - People with intellectual disabilities in iceland: a bourdieuean interpretation of self-advocacy. AB - There are many barriers to social participation in Iceland for people with intellectual disabilities. This article builds on qualitative research with young adults with intellectual disabilities. The purpose of this article is to develop an approach where the struggles over the meaning of social participation of people with intellectual disabilities are seen as social strategies. In the article, the authors suggest that people with intellectual disabilities are carving out a space where intellectual disability is gaining higher social status. They also posit that people with intellectual disabilities use several social strategies in the emerging field of self-advocacy for the purpose of improving their social position. Thus, the article contributes to a new social understanding of disability and how people with disabilities gain authority over their lives and experiences. PMID- 20020800 TI - Residential characteristics, social factors, and mortality among adults with intellectual disabilities: transitions out of nursing homes. AB - The present study examined the degree to which residential characteristics and social factors are associated with mortality, after controlling for personal characteristics, among adults with intellectual disabilities who have resided in nursing homes (facilities providing skilled care and related services) at baseline in the Chicago area. Initial assessments were conducted on 330 residents, and 2 follow ups were performed over 10 years. Seventy-one residents had died by the end of the study. The variables examined included personal characteristics of age, gender, intellectual level, health, and adaptive behavior; residential characteristics of type of residence, size of facility, attractiveness of physical environment (cleanliness, conditions, and aesthetic appeal), and diversity of physical environment (personalization of residents' rooms and distinctiveness of all living spaces); and social factors of family involvement and community integration. The results indicated, beyond age, adaptive behavior, and health, that size of residences, residential characteristics, and social factors are related to mortality. PMID- 20020801 TI - Praying with lior (trachtman, 2007): the myth or reality of intellectual disability. PMID- 20020802 TI - Casting call for a supporting role. PMID- 20020803 TI - Challenging understandings of "theory of mind": a brief report. PMID- 20020806 TI - A new species of Cosmocercoides (Nematoda: Cosmocercidae) from a gymnophthalmid lizard of western Brazil. AB - A Cosmocercoides sauria n. sp., an intestinal parasite of the gymnophthalmid lizard Iphisa elegans from western Brazil, is described. Of the 18 species previously described, C. sauria represents the 19th species, and is the third to be reported in the Neotropical region. Moreover, C. sauria is the second species reported from a reptilian host and is distinguished by a smaller number of rosette papillae (4 pairs). Additionally, the new species can be distinguished from the other Neotropical species (Cosmocercoides lilloi Ramallo, Bursey and Goldberg, 2007 and C. variabilis (Harwood, 1930)) by possessing smaller spicule size and presence of both gubernaculum and lateral alae in Cosmocercoides lilloi . PMID- 20020807 TI - Antibodies to Toxoplasma gondii in stray and household dogs in Guangzhou, China. AB - The frequency of Toxoplasma gondii antibodies in stray and household dogs in Guangzhou, China was examined by ELISA on serum samples from 150 animals (36 strays and 114 from households) and the overall prevalence was 21.3%. The extent of infection in stray dogs (33.3%) was significantly higher (P < 0.05) than in household dogs (17.5%). Infection in male and female dogs of both groups was not significantly different (P >or= 0.05), i.e., 31.8% versus 35.7% for male and female in stray dogs, and 14.5% versus 22.2% in household dogs. The results of the present investigation indicate that the seroprevalence of T. gondii infection in dogs was high in Guangzhou, especially in strays. Therefore, it is essential to implement integrated strategies to prevent and control T. gondii infection in both stray and household dogs. PMID- 20020808 TI - Prevalence of antibodies to Trypanosoma cruzi, Leishmania infantum, Encephalitozoon cuniculi, Sarcocystis neurona, and Neospora caninum in Capybara, Hydrochoerus hydrochaeris, from Sao Paulo State, Brazil. AB - Little is known about the importance of capybara, Hydrochoerus hydrochaeris, as reservoirs for parasites of zoonotic or veterinary importance. Sera from 63 capybaras, from 6 counties in the state of Sao Paulo, Brazil, were examined for antibodies to Trypanosoma cruzi, Leishmania infantum, Encephalitozoon cuniculi, Sarcocystis neurona, and Neospora caninum using an indirect immunofluorescent antibody test. Five (8%) of the 63 capybaras had antibodies to T. cruzi epimastigotes. None of the samples from capybara reacted positively with L. infantum promastigotes or with spores of E. cuniculi . Two (3%) of the serum samples were positive for antibodies to S. neurona merozoites, and 2 (3%) of the serum samples were positive for antibodies to N. caninum tachyzoites. A serum sample from 1 capybara was positive for antibodies to both T. cruzi and N. caninum. None of the remaining 62 samples reacted with more than 1 parasite. PMID- 20020809 TI - Public health nurses in rural/frontier one-nurse offices. AB - INTRODUCTION: Public health nursing is the foundation of the United States' (US) public health system, particularly in rural and remote areas. Recent increasing interest in public health in the USA has highlighted that there is limited information available about public health nursing in the most isolated areas, particularly in the US. The purposes of this study were to: (1) describe the characteristics, competency levels, and practice patterns of public health nurses (PHNs) working in remote one-nurse offices; and (2) compare PHNs working in one nurse offices with nurses working in multi-nurse offices in Idaho, in relation to their demographic characteristics, practice patterns and competency levels. METHODS: Using a cross-sectional descriptive design, a statewide sample of 124 PHNs in Idaho, including 15 working in one-nurse satellite offices, were assessed in relation to their demographic characteristics, experience, educational background, job satisfaction, practice characteristics, and competency levels in March to May 2007. RESULTS: The solo (nurses working in one-nurse offices) PHNs were based in 15 different counties, 10 frontier (population density of less than 7 persons/1.6 km(2); 7 persons/mile(2)) and 5 rural. The counties ranged in population from 2781 to 28 114 (mean = 11 013), with population densities ranging from 0.9 to 29.4 persons/1.6 km(2) (mean = 8.6; 0.9 to 29.4 persons/mile(2)). The distance from their offices to the district main office ranged from 25.8 to 241.4 km (mean = 104 km; 16 to 150 miles, mean = 64.6 miles). All the solo PHNs were Caucasian females, with a mean age of 46.9 years and a mean of 22.5 years' nursing experience. Educationally, 7 (47%) held a bachelor degree in nursing, 6 (40%) had associates degrees, 1 (7%) had a diploma in nursing, and 1 (7%) was a licensed practical nurse (LPN). These solo PHNs provided a wide array of services with support from other nurses in the district, including epidemiology, family planning/sexually transmitted disease clinics, immunization clinics, communicable disease surveillance, and school nursing. They expressed strong job satisfaction, citing the benefits of autonomy, variety, and close community ties, but also voiced some frustrations related to isolation. Their self-rated levels of competency were highest in the areas of communication, cultural competency, community dimensions of care, and leadership/systems thinking skills; and lowest in the areas of financial management, analytical assessment, policy development/program planning, and basic public health sciences skills. When the solo PHNs were compared with PHNs based in multi-nurse offices, there were no statistically significant differences between the solo and non-solo PHNs in demographics or competency levels, except in the competency area of community dimensions of practice skills. The mean self-rating for solo PHNs in relation to community dimensions of practice skills was significantly higher (3.9) than non solo PHNs (3.2) (t = 3.547, p = .002). CONCLUSIONS: These findings suggest that US PHNs practicing in isolated one-nurse offices in rural and remote communities are comparable to PHNs working in less isolated settings; however, solo nurses may have stronger community dimensions of practice skills. Their practice is more generalized than other PHNs and they express high levels of job satisfaction. The study was limited in that it was conducted in only one state and data were collected only by self-report. Further research is indicated to describe this unique subset of PHNs, particularly in terms of factors promoting recruitment and retention. Additional study into the conceptual aspect of isolation is also indicated in relation to public health practice in rural and remote areas. PMID- 20020810 TI - First molecular detection of Anaplasma phagocytophilum in European brown bear (Ursus arctos). AB - In Europe, Anaplasma phagocytophilum circulates in natural foci in a tick-host cycle. Up to now, antibodies against A. phagocytophilum as well as pathogen's DNA were recorded in several domestic and wild animals. Nevertheless, the reservoir host range is still under investigation. Tissue samples from European brown bears (Ursus arctos) were tested for the presence of A. phagocytophilum DNA by a PCR amplification of 16S rRNA gene. The results of our study provides the evidence, that the range of animals involved in the circulation and maintenance of A. phagocytophilum in natural foci, is extended of another ursine carnivore, European brown bear (Ursus arctos). PMID- 20020811 TI - A case of person-to-person transmission of Q fever from an active duty serviceman to his spouse. AB - Coxiella burnetii has recently gained military relevance given its potential as a bioterrorism agent, and the multiple cases reported among U.S. military personnel deployed to the Middle East. Sexual transmission of Q fever is rare but has been reported in the literature. We describe the possible sexual transmission of Q fever from a returning serviceman from Iraq to his wife. In a recent editorial commentary, Dr. Raoult wrote about the reemergence of Q fever after September 11, 2001 (Raoult 2009). Indeed, C. burnetii has gained military relevance given its potential as a bioterrorism agent and the multiple cases reported among military personnel deployed in Southwest/Central Asia and North Africa (Botros et al. 1995 , Meskini et al. 1995 , Leung-Shea and Danaher 2006 ). Human serosurveys in these geographic areas have reported prevalence rates for Q fever ranging from 10% to 37% in contrast to the United States, which has an estimated Q fever seroprevalence of 3.1% (Botros et al. 1995, Meskini et al. 1995, Anderson et al. 2009). There is no data available for Q fever seroprevalence in Iraq. As a consequence, native populations in these regions may be more likely to possess immunity, and newcomers, such as U.S. military personnel, would be vulnerable to acute infection (Derrick 1973). We report on the possible sexual transmission of C. burnetii from a serviceman in the late recovery of acute Q fever to his wife. PMID- 20020812 TI - Time series analysis of the impact of oral vaccination on raccoon rabies in West Virginia, 1990-2007. AB - To assess the potential impact of oral rabies vaccination (ORV) on the occurrence of raccoon rabies in the mid-Atlantic region, temporal and seasonal trends of raccoon rabies cases reported in West Virginia from 1990 to 2007 were identified with both descriptive statistical analysis and exploratory time series analysis. Raccoon rabies cases in the non-ORV region maintain an enzootic pattern and increase over time; a bimodal seasonal pattern is observed with a large peak in April and a smaller peak in August. The results of the model indicate that the effect of the ORV intervention to control raccoon rabies was statistically significant. ORV should be attempted in other enzootic raccoon rabies areas. PMID- 20020813 TI - Potential zoonotic pathways of Salmonella enteritidis in laying farms. AB - Salmonella Enteritidis is a communicable zoonotic bacterium. The present investigation was done to evaluate the potential occurrence of Salmonella Enteritidis in laying hen farms and its contamination pathways. Samples were collected from 10 laying hen farms located in the Delta of Egypt. Cloacal swabs (n=300), eggshell swabs (n=400), and hand swabs from egg packagers (n=38) were collected. Pools of ovary and oviduct were obtained from 150 hens; all samples were examined for the presence of Salmonella Enteritidis. Results indicated that Salmonella Enteritidis may initially occupy in ovary, oviduct, and cloaca. The risk for eggshell contamination is highly impending from laying hen flocks infected with Salmonella Enteritidis with percentage of >30%. Farms having eggshell contamination percentage of >60% with Salmonella Enteritidis provided a high risk for packagers' hand contamination. Questionnaire of egg packagers specified that seven out of the eight smoker packagers suffered from repeated Salmonellosis. Thus, smoking during egg packing process could be considered as an exposure factor to contract the infection via hand-cigarette-mouth route. PMID- 20020814 TI - Prevalence of five pathogenic agents in questing Ixodes ricinus ticks from western France. AB - In Europe, Ixodes ricinus ticks are vectors of many emerging pathogens, including Borrelia burgdorferi sensu lato (sl), Anaplasma phagocytophilum, spotted fever group Rickettsia sp., Babesia sp., and very likely Bartonella sp. In this study, we looked for the presence of DNA of these microorganisms in 572 ticks from two forests in the west of France. DNA extraction and polymerase chain reaction (PCR) amplification were performed on individual nymphal, male, and female I. ricinus ticks. Amplification from 1 tick among the 572 samples (0.2%) resulted in PCR products with Bartonella-specific primers. Sequence analysis of the amplified fragment did not lead to species identification. Two ticks (0.3%) carried A. phagocytophilum-specific DNA. Eight ticks (1.4%) were positive with spotted fever group Rickettsia-specific primers, and all PCR fragments were related to Rickettsia helvetica. Thirty-five ticks (6.1%) were positive with B. burgdorferi sl-specific primers; the sequences were all related to Borrelia garinii or Borrelia afzelii, except one that was related to Borrelia carolinensis, a newly described species never reported in Europe so far. Thirty-five ticks (6.1%) carried Babesia sp. DNA. Female adults were more infected by B. burgdorferi sl than male adults. The prevalence of B. burgdorferi sl and Babesia sp. was significantly different between the two forests, with a higher prevalence of B. burgdorferi sl in ticks from the forest of Prince and a higher prevalence of Babesia sp. in ticks from the forest of Gavre. To our knowledge, this is the first study that has detected all five pathogens in questing I. ricinus in the west of France and the first report of B. carolinensis DNA in ticks in Europe. PMID- 20020815 TI - Seroprevalence of Trypanosoma cruzi among eleven potential reservoir species from six states across the southern United States. AB - Trypanosoma cruzi, the causative agent of Chagas' disease, is a substantial public health concern in Latin America. Although rare in humans and domestic animals in the United States, T. cruzi is commonly detected in some wildlife species, most commonly raccoons (Procyon lotor) and Virginia opossums (Didelphis virginiana). To increase our understanding of the reservoir host species range and geographic distribution, 11 species of mammals from six states spanning the known range of T. cruzi (Arizona, California, Florida, Georgia, Missouri, and Virginia) were tested for antibodies to T. cruzi using indirect immunofluorescent antibody testing. In addition, culture isolation attempts were conducted on a limited number of animals from Georgia and Florida. Evidence of T. cruzi was found in every state except California; however, low numbers of known reservoirs were tested in California. In general, the highest seroprevalence rates were found in raccoons (0-68%) and opossums (17-52%), but antibodies to T. cruzi were also detected in small numbers of striped skunks (Mephitis mephitis) from Arizona and Georgia, bobcats (Lynx rufus) from Georgia, two coyotes (Canis latrans) from Georgia and Virginia, and a ringtail (Bassariscus astutus) from Arizona. Culture based prevalence rates for raccoons were significantly greater than those for opossums; however, seroprevalences of raccoons and opossums from several geographic locations in Georgia and Florida were not different, indicating that exposure rates of these two species are similar within these areas. For both raccoons and opossums, seroprevalence was significantly higher in females than in males. No difference was detected in seroprevalence between adults and juveniles and between animals caught in urban and rural locations. Our results indicate that T. cruzi prevalence varies by host species, host characteristics, and geographic region and provides data to guide future studies on the natural history of T. cruzi in the United States. PMID- 20020817 TI - Liposomes modified with YIGSR peptide for tumor targeting. AB - YIGSR peptide anchored sterically stabilized liposomes (YIGSR-SL) were investigated for selective and preferential presentation of carrier contents at angiogenic endothelial cells overexpressing laminin receptors on and around tumor tissue and thus for assessing their targetabilty. In vitro endothelial cell binding of liposomes exhibited 7-fold higher binding of YIGSR-SL to HUVEC in comparison to the nontargeted sterically stabilized liposomes (SL). Spontaneous lung metastasis and angiogenesis assays show that YIGSR peptide anchored liposomes are significantly (P or=10 nmol/L. When ELISAs were introduced on fully automated test systems to measure neopterin concentrations, results were generally higher and the cut-off was adjusted up to 12 nmol/L. METHODS: In early 2006, new equipment (Hamilton Microlab Star pipettor + Dade Behring ELISA Processor III, BEP III) became available which monitors sample and reagent volumes for each uptake and dispense, and takes the viscosity of the fluids into account. Using this system, the performance of neopterin determinations was evaluated over 21 months and compared to earlier data. RESULTS: When the new apparatuses were introduced, an immediate decrease in mean neopterin concentrations and the percentage of samples with neopterin concentrations >or=10 nmol/L was observed. The 55,516 measurements performed in 2007 showed a mean value of 5.5 nmol/L. Neopterin in 680 donations (1.23%) were >or=10 nmol/L. CONCLUSIONS: The new Hamilton diluter increased the accuracy of the neopterin test procedure. Results are now identical with those of RIA and ELISA when run in a semi-automated manner. Differences in pipetting performance of standards and samples are considered as one possible explanation for the earlier discrepancies seen with the automated apparatus. PMID- 20020819 TI - Integrated PCR amplification and detection processes on a Lab-on-Chip platform: a new advanced solution for molecular diagnostics. AB - BACKGROUND: Several microdevices have been developed to perform only a single step of a genotyping process, such as PCR or detection by probe hybridization. Here, we describe a Lab-on-Chip (LoC) platform integrating a PCR amplification microreactor with a customable microarray for the detection of sequence variations on human genomic DNA. METHODS: Preliminary work was focused on developing the single analytical steps including PCR and labeling strategies of the amplified product by conventional reference systems. The optimized protocols included a 1:4 forward:reverse primer ratio for asymmetric PCR, and Cy5-dCTP multiple incorporation for the generation of a labeled PCR product to be hybridized to complementary probes bound to the chip surface. RESULTS: Final conditions were applied to the fully integrated LoC platform for the detection of the IVSI-110 G > A mutation in the human beta-globin (HBB) gene associated with beta-thalassemia, used as a model of genetic application, allowing for correct genotyping of 25 samples that were heterozygous, homozygous or wild-type for this mutation. CONCLUSIONS: The overall results show that the present platform is very promising for rapid identification of DNA sequence variations in an integrated, cost effective and convenient silicon chip format. PMID- 20020820 TI - Multicentre evaluation of the Tosoh HbA1c G8 analyser. AB - BACKGROUND: We report a Dutch-Belgian multicentre evaluation of the Tosoh HLC 723G8 glycohaemoglobin analyser, an ion-exchange HPLC instrument for the separation and quantification of haemoglobin A1c (HbA1c) in whole blood. METHODS: We evaluated the analytical performances of the Tosoh G8 analyser and compared the results for blood samples with its predecessor, the Tosoh G7, and with two other widely used analysers, the Bio-Rad Variant II and Adams Arkray HA-8160. RESULTS: Within- and between-batch imprecision [coefficient of variation (CVs)] was <0.5% and 2%, respectively, and compared favourably with the G7. The excellent performances in terms of speed (1.6 min/analysis) did not result in increased variability of the results or carry-over between samples. The method shows no interference from carbamylated haemoglobin, and recognises the presence of haemoglobinopathies, which triggers the correction of the HbA1c result. Comparison with established methods showed good correlation, not only with the G7 but also with the Variant II and HA-8160 systems. CONCLUSIONS: With respect to reproducibility, chromatographic resolution, speed of analysis and identification of Hb variants, the Tosoh G8 analyser can be considered to be state of the art. PMID- 20020821 TI - Decrease in uric acid in acute ischemic stroke correlates with stroke severity, evolution and outcome. AB - BACKGROUND: Although uric acid (UA) is one of the most important antioxidants in plasma and appears to be neuroprotective in animal models, results from human studies are controversial. In this study, we investigated the kinetics of serum UA concentrations in the acute, subacute and chronic phase of ischemic stroke and its relation with initial stroke severity, stroke evolution in the subacute phase and long-term stroke outcome. METHODS: Serum concentrations of UA were measured in 199 stroke patients at admission (median, 2.8 h after stroke onset), at 24 h, 72 h, day 7, month 1 and month 3 after onset of stroke. We evaluated the relationship between changes in UA concentrations and (a) stroke severity [patients with transient ischemic attack (TIA) vs. stroke patients, National Institutes of Health Stroke Scale (NIHSS) score at admission], (b) stroke evolution (stroke progression, infarct volume at 72 h), and (c) stroke outcome [modified Rankin scale (mRS) score at month 3, mortality]. RESULTS: UA concentrations decreased significantly during the first 7 days after stroke onset before returning to baseline (p < 0.001). Mean plasma UA concentrations decreased from 336.66 +/- 113.01 micromol/L at admission to 300.37 +/- 110.04 micromol/L at day 7 (p < 0.001) in patients with stroke, but did not change significantly in patients with TIA. Changes in UA concentrations from admission to day 7 (DeltaUA(day 7)) correlated with the NIHSS score (rho = 0.32; p < 0.001), stroke progression (rho = 0.29; p = 0.001), infarct volume (rho = 0.37; p < 0.001), mRS score (rho = 0.28; p = 0.001) and mortality (p = 0.010). CONCLUSIONS: Decreases in UA during the first week after onset of stroke correlates with more severe stroke, unfavorable stroke evolution, and poor long-term stroke outcome. PMID- 20020822 TI - Gender does not influence cystatin C concentrations in healthy volunteers. AB - BACKGROUND: Current data are conflicting about the influence of gender on cystatin C concentrations. The goal of this study was to determine the reference interval for serum cystatin C in normal Brazilian subjects, taking into account the influence of gender. METHODS: Ninety-seven healthy volunteers, aged 18-70 years, 44% male, had glomerular filtration rate (GFR) measured using a (51)Cr EDTA method and estimated with the Modification of Diet in Renal Disease (MDRD) study equation. Serum cystatin C was measured using a turbidimetric method, and creatinine by the Jaffe method. RESULTS: Mean serum cystatin C was not significantly different between males and females, 0.62 +/- 0.12 vs. 0.65 +/- 0.12 mg/L, respectively (p = 0.26). However, median serum creatinine was significantly higher in men [97 (80-115) vs. 80 (53-88) micromol/L; p = 0.0001]. There were also no significant differences between genders with respect to GFR measured using (51)Cr-EDTA (103 +/- 14 for males and 106 +/- 19 mL/min/1.73 m(2) for females, p = 0.47), and estimated with the MDRD equation (86 +/- 12 vs. 83 +/ 16 mL/min/1.73 m(2), respectively, p = 0.24). CONCLUSIONS: There was no effect of gender on serum cystatin C, as well as on measured and estimated GFR. PMID- 20020823 TI - Nasal administration of ondansetron using a novel microspheres delivery system Part II: ex vivo and in vivo studies. AB - Gellan gum-based mucoadhesive microspheres of ondansetron hydrochloride for intranasal systemic delivery were prepared to avoid first pass effect, an alternative route of administration to injection and to enhance systemic bioavailability of ondansetron hydrochloride. The microspheres were prepared using spray method. The evaluation results of microspheres were reported in our previous study. The aim of this work was to study the in vivo performance of mucoadhesive microspheres in comparison with oral and intravenous preparations of ondansetron hydrochloride. The nasal delivery system gave increased AUC(0-240) and C(max) as compared to those of oral delivery. In conclusion, the gellan gum based microsphere formulation of ondansetron hydrochloride with mucoadhesive properties with increased permeation rate is promising for prolonging nasal residence time and thereby nasal absorption. PMID- 20020824 TI - Developing an asymptomatic mucosal herpes vaccine: the present and the future. PMID- 20020826 TI - Phosphorylation of paramyxovirus phosphoprotein and its role in viral gene expression. AB - Paramyxoviruses include many important human and animal pathogens such as measles virus, mumps virus, human parainfluenza viruses, and respiratory syncytial virus, as well as emerging viruses such as Nipah virus and Hendra virus. The paramyxovirus RNA-dependent RNA polymerase consists of the phosphoprotein (P) and the large protein. Both of these proteins are essential for viral RNA synthesis. The P protein is phosphorylated at multiple sites, probably by more than one host kinase. While it is thought that the phosphorylation of P is important for its role in viral RNA synthesis, the precise role of P protein phosphorylation remains an enigma. For instance, it was demonstrated that the putative CKII phosphorylation sites of the P protein of respiratory syncytial virus play a role in viral RNA synthesis using a minigenome replicon system; however, mutating these putative CKII phosphorylation sites within a viral genome had no effect on viral RNA synthesis, leading to the hypothesis that P protein phosphorylation, at least by CKII, does not play a role in viral RNA synthesis. Recently, it has been reported that the phosphorylation state of the P protein of parainfluenza virus 5, a prototypical paramyxovirus, correlates with the ability of P protein to synthesize viral RNA, indicating that P protein phosphorylation does in fact play a role in viral RNA synthesis. Furthermore, host kinases PLK1, as well as AKT1 have been found to play critical roles in paramyxovirus RNA synthesis through regulation of P protein phosphorylation status. Beyond furthering our understanding of paramyxovirus RNA replication, these recent discoveries may also result in a new paradigm in treating infections caused by these viruses, as host kinases that regulate paramyxovirus replication are investigated as potential targets of therapeutic intervention. PMID- 20020827 TI - Mucosal treatments for herpes simplex virus: insights on targeted immunoprophylaxis and therapy. AB - Herpes simplex virus (HSV) serotypes 1 and 2 establish lifelong infections that can produce reactivated pools of virus at mucosal sites where primary infections were initiated. No approved vaccines are available. To break the transmission cycle, interventions must either prevent infection or reduce infectivity at mucosal sites. This article discusses the recent experimental successes of immunoprophylactic and therapeutic compounds that enhance resistance and/or reduce viral loads at genital and ocular mucosa. Current data indicate Toll-like receptor agonists and selected immunomodulating compounds effectively increase the HSV infection threshold and hold promise for genital prophylaxis. Similarly, immunization at genital and extragenital mucosal sites is discussed. Finally, preclinical success with novel immunotherapies for ocular HSV that address herpetic keratitis and corneal blindness is reviewed. PMID- 20020830 TI - Molecular phylogenetics and epidemiology of Candida albicans. AB - Candida albicans, a diploid yeast commensal and opportunist pathogen, has evolved unusual mechanisms for maintenance of genetic diversity in the absence of a complete sexual cycle. These include chromosomal polymorphisms, mitotic recombination events, and gains and losses of heterozygosity, superimposed on a fundamentally clonal mode of reproduction. Molecular typing of C. albicans strains shows geographical evolutionary associations but these have become partially blurred, probably as a result of extensive human travel. Individual patients usually carry a single C. albicans strain type, but this may undergo microvariation leading to detection of mixtures of closely related types. Associations have been found between clade 1, the most common multilocus sequence typing cluster of related C. albicans strains, and resistance to flucytosine and terbinafine. There are also clade-related associations with lengths of tandem repeats in some cell-surface proteins, but not with virulence or type of infection. PMID- 20020828 TI - Innate immune evasion strategies of influenza viruses. AB - Influenza viruses are globally important human respiratory pathogens. These viruses cause seasonal epidemics and occasional worldwide pandemics, both of which can vary significantly in disease severity. The virulence of a particular influenza virus strain is partly determined by its success in circumventing the host immune response. This article briefly reviews the innate mechanisms that host cells have evolved to resist virus infection, and outlines the plethora of strategies that influenza viruses have developed in order to counteract such powerful defences. The molecular details of this virus-host interplay are summarized, and the ways in which research in this area is being applied to the rational design of protective vaccines and novel antivirals are discussed. PMID- 20020831 TI - Malaria parasite mutants with altered erythrocyte permeability: a new drug resistance mechanism and important molecular tool. AB - Erythrocytes infected with plasmodia, including those that cause human malaria, have increased permeability to a diverse collection of organic and inorganic solutes. While these increases have been known for decades, their mechanistic basis was unclear until electrophysiological studies revealed flux through one or more ion channels on the infected erythrocyte membrane. Current debates have centered on the number of distinct ion channels, which channels mediate the transport of each solute and whether the channels represent parasite-encoded proteins or human channels activated after infection. This article reviews the identification of the plasmodial surface anion channel and other proposed channels with an emphasis on two distinct channel mutants generated through in vitro selection. These mutants implicate parasite genetic elements in the parasite-induced permeability, reveal an important new antimalarial drug resistance mechanism and provide tools for molecular studies. We also critically examine the technical issues relevant to the detection of ion channels by electrophysiological methods; these technical considerations have general applicability for interpreting studies of various ion channels proposed for the infected erythrocyte membrane. PMID- 20020829 TI - Current understanding of Pneumocystis immunology. AB - Pneumocystis jirovecii is the opportunistic fungal organism that causes Pneumocystis pneumonia (PCP) in humans. Similar to other opportunistic pathogens, Pneumocystis causes disease in individuals who are immunocompromised, particularly those infected with HIV. PCP remains the most common opportunistic infection in patients with AIDS. Incidence has decreased greatly with the advent of HAART. However, an increase in the non-HIV immunocompromised population, noncompliance with current treatments, emergence of drug-resistant strains and rise in HIV(+) cases in developing countries makes Pneumocystis a pathogen of continued interest and a public health threat. A great deal of research interest has addressed therapeutic interventions to boost waning immunity in the host to prevent or treat PCP. This article focuses on research conducted during the previous 5 years regarding the host immune response to Pneumocystis, including innate, cell-mediated and humoral immunity, and associated immunotherapies tested against PCP. PMID- 20020835 TI - Wound healing after laser skin resurfacing: the effect of a silver sulfadiazine hyaluronic acid-containing cream under an occlusive dressing. AB - INTRODUCTION: Occlusive dressings promote wound healing after laser skin resurfacing. Our goal was to evaluate whether a cream containing hyaluronic acid silver sulfadiazine could improve this process. METHODS: Ten guinea pigs received laser resurfacing on the skin of their backs. Polyurethane foam film adhesive dressing was applied. A cream containing hyaluronic acid-silver sulfadiazine was applied under the dressing over half of the wound. Skin biopsies were taken at days 2, 4, 7 and 9 and examined by a pathologist blinded to the study. RESULTS: Re-epithelialization appeared to occur faster in the cream-treated areas, together with a lower level of acute inflammation in the early phase of the healing process. CONCLUSIONS: The cream containing silver sulfadiazine/hyaluronic acid seems to improve wound healing after laser skin resurfacing in guinea pigs. The clinical application of this finding on humans should be assessed. PMID- 20020832 TI - Defensins as anti-inflammatory compounds and mucosal adjuvants. AB - Human neutrophil peptide alpha-defensins and human beta-defensins are small, well characterized peptides with broad antimicrobial activities. In mixtures with microbial antigens, defensins attenuate proinflammatory cytokine responses by dendritic cells in culture, attenuate proinflammatory cytokine responses in the nasal fluids of exposed mice and enhance antibody responses in the serum of vaccinated mice. Although the exact mechanisms are unknown, defensins first start by binding to microbial antigens and adhesins, often attenuating toxic or inflammatory-inducing capacities. Binding is not generic; it appears to be both defensin-specific and antigen-specific with high affinities. Binding of defensins to antigens may, in turn, alter the interaction of antigens with epithelial cells and antigen-presenting cells attenuating the production of proinflammatory cytokines. The binding of defensins to antigens may also facilitate the delivery of bound antigen to antigen-presenting cells in some cases via specific receptors. These interactions enhance the immunogenicity of the bound antigen in an adjuvant-like fashion. Future research will determine the extent to which defensins can suppress early events in inflammation and enhance systemic antibody responses, a very recent and exciting concept that could be exploited to develop therapeutics to prevent or treat a variety of oral mucosal infections, particularly where inflammation plays a role in the pathogenesis of disease and its long-term sequelae. PMID- 20020836 TI - Periorbital skin tightening with a broadband infrared device: preliminary study results. AB - Periorbital rhytids are often the first signs of aging skin for which young patients seek a non-invasive, no downtime treatment. Recently, several skin tightening modalities based on deep dermal heating with various energy sources, such as monopolar and bipolar radiofrequency, as well as laser and broadband infrared light sources, have been introduced to treat mild wrinkles and skin laxity. This report presents preliminary study results using a new infrared device for the treatment of periorbital rhytids on 11 volunteer patients. PMID- 20020833 TI - Defensins attenuate cytokine responses yet enhance antibody responses to Porphyromonas gingivalis adhesins in mice. AB - AIM: Our aim is to assess the ability of human neutrophil peptide alpha-defensins (HNPs) and human beta-defensins (HBDs) to attenuate proinflammatory cytokine responses and enhance antibody responses to recombinant hemagglutinin B (rHagB) or recombinant fimbrillin A (rFimA) from Porphyromonas gingivalis 381 in mice. MATERIALS & METHODS: In the first study, C57BL/6 mice were given 10 microg rHagB or rFimA without and with 1 microg HNP1, HNP2, HBD1, HBD2 or HBD3. At 24 h, mice were euthanized and cytokine concentrations were determined in nasal wash fluid (NWF), bronchoalveolar lavage fluids, saliva and serum. In the second study, C57BL/6 mice were given 10 microg rHagB or rFimA without and with 1 microg HNPs or HBDs similarly on days 0, 7 and 14. At 21 days, mice were euthanized and rHagB and rFimA-specific antibody responses were determined in NWF, bronchoalveolar lavage fluids, saliva and serum. RESULTS: Mice given rHagB + HNP2, rHagB + HBD1 and rHagB + HBD3 produced significantly lower (p < 0.05) IL-6 responses than mice given rHagB alone. Mice given rHagB + HNP1, rHagB + HNP2, rHagB + HBD1 and rHagB + HBD3 produced significantly lower (p < 0.05) keratinocyte-derived chemokine responses than mice given rHagB alone. Mice given rFimA produced very low levels of IL-6 and only moderate levels of keratinocyte-derived chemokine in NWF that were not attenuated by prior incubation of rFimA with any defensin. Mice given rHagB + HNP1 produced a significantly higher (p < 0.05) serum IgG antibody response than mice given rHagB alone and mice given rFimA + HNP2 produced a higher, but not significant, antibody response. CONCLUSION: The ability of HNPs and HBDs to attenuate proinflammatory cytokine responses in murine NWF and enhance IgG antibody responses in serum was dependent upon both the defensin and antigen of P. gingivalis. PMID- 20020837 TI - Unwanted facial hair removal with laser treatment improves quality of life of patients. AB - BACKGROUND: Unwanted facial hair can have adverse psychological effects on women and reduce their quality of life. OBJECTIVE: To assess the effects of unwanted facial hair removal with laser on improving quality of life. METHODS: In this study, 70 patients treated for unwanted facial hair by laser were assessed by Dermatology Life Quality Index (DLQI) questionnaire on admission and 3 months later after three sessions of laser treatment. RESULTS: The DLQI score before treatment was 9.42 +/- 5.99, which was reduced to 3.12 +/- 3.40 after laser treatment (p <0.05). CONCLUSION: Unwanted facial hair removal with laser can improve the quality of life of patients. PMID- 20020838 TI - The value of a postoperative computed tomography scan. PMID- 20020839 TI - Wrong-site craniotomy. PMID- 20020840 TI - The clinical significance and optimal timing of postoperative computed tomography following cranial surgery. AB - OBJECT: This study was conducted to evaluate the value of postoperative CT scans in determining the probability of return to the operating room (OR) and the optimal time to obtain such scans to determine the effects of surgery. METHODS: Between January and December 2006 (12 months), all postoperative head CT scans obtained for 3 individual surgeons were reviewed. Scans were divided into 3 groups, which were determined by the preference of each surgeon: Group A (early scans-scheduled between 0 and 7 hours); Group B (delayed scans-scheduled between 8 and 24 hours); and Group C (urgent scans-ordered because of a new neurological deficit). The initial scans were reviewed and analyzed in 2 different fashions. The first was to analyze the efficacy of the scans in predicting return to the OR. The second was to determine the optimal time for obtaining a scan. The second analysis was a review of serial postoperative scans for expected versus unexpected findings and changes in the acuity of these findings over time. RESULTS: In 251 (74%) of 338 cases, the patients had postoperative head CT scans within 24 hours of surgery. Analysis 1 determined the percent of patients returning to the OR for emergency treatment based on postoperative scans: Group A (early)-133 patients, with 0% returning to the OR; Group B (delayed)-108 patients, with 0% returning to the OR; and Group C (urgent)-10 patients, with 30% returning to the OR (p < 0.05). Analysis 2 determined the optimal timing of postoperative scans and changes in scan acuity: Group A (early scan) had an 11% incidence of change in acuity on subsequent scans. Group B (delayed scan) had a 3% incidence of change in acuity on follow-up scans (p < 0.05). CONCLUSIONS: Routine postoperative scans at 0-7 hours or at 8-24 hours are not predictive of return to the OR, whereas patients with a new neurological deficit in the postoperative period have a 30% chance of emergency reoperation based on CT scans. In addition, early postoperative scans (0-7 hours) fail to predict CT changes, which might evolve over time and may influence postoperative medical management. PMID- 20020841 TI - Poor drug distribution as a possible explanation for the results of the PRECISE trial. AB - OBJECT: Convection-enhanced delivery (CED) is a novel intracerebral drug delivery technique with considerable promise for delivering therapeutic agents throughout the CNS. Despite this promise, Phase III clinical trials employing CED have failed to meet clinical end points. Although this may be due to inactive agents or a failure to rigorously validate drug targets, the authors have previously demonstrated that catheter positioning plays a major role in drug distribution using this technique. The purpose of the present work was to retrospectively analyze the expected drug distribution based on catheter positioning data available from the CED arm of the PRECISE trial. METHODS: Data on catheter positioning from all patients randomized to the CED arm of the PRECISE trial were available for analyses. BrainLAB iPlan Flow software was used to estimate the expected drug distribution. RESULTS: Only 49.8% of catheters met all positioning criteria. Still, catheter positioning score (hazard ratio 0.93, p = 0.043) and the number of optimally positioned catheters (hazard ratio 0.72, p = 0.038) had a significant effect on progression-free survival. Estimated coverage of relevant target volumes was low, however, with only 20.1% of the 2-cm penumbra surrounding the resection cavity covered on average. Although tumor location and resection cavity volume had no effect on coverage volume, estimations of drug delivery to relevant target volumes did correlate well with catheter score (p < 0.003), and optimally positioned catheters had larger coverage volumes (p < 0.002). Only overall survival (p = 0.006) was higher for investigators considered experienced after adjusting for patient age and Karnofsky Performance Scale score. CONCLUSIONS: The potential efficacy of drugs delivered by CED may be severely constrained by ineffective delivery in many patients. Routine use of software algorithms and alternative catheter designs and infusion parameters may improve the efficacy of drugs delivered by CED. PMID- 20020842 TI - Cushing's lost cases of "radium bomb" brachytherapy for gliomas. AB - Although recent efforts to advance the treatment of gliomas through radiotherapy and chemotherapy may seem to be a relatively new area of growth and development, these efforts have been in progress since the therapeutic potential of radiation therapy was discovered in the late 19th century. Cushing's use of brachytherapy has been mentioned several times in the literature without receiving an appropriate in-depth analysis. The reasoning behind Cushing's initial use of brachytherapy was not fully examined, and a close analysis of the outcomes of this therapy was not made. In addition, Cushing's use of his "radium bomb" occurred more commonly than the 3 cases previously documented. The authors reviewed all the patient records available at the Cushing Brain Tumor Registry- which represents the most complete series of patient records from the Cushing era -and selected those patients who underwent treatment with Cushing's "radium bomb." The authors place these early attempts to optimize interstitial radiation of brain tumors in their historical perspective. PMID- 20020844 TI - Use of an artificial neural network to predict head injury outcome. AB - OBJECT: The authors describe the artificial neural network (ANN) as an innovative and powerful modeling tool that can be increasingly applied to develop predictive models in neurosurgery. They aimed to demonstrate the utility of an ANN in predicting survival following traumatic brain injury and compare its predictive ability with that of regression models and clinicians. METHODS: The authors designed an ANN to predict in-hospital survival following traumatic brain injury. The model was generated with 11 clinical inputs and a single output. Using a subset of the National Trauma Database, the authors "trained" the model to predict outcome by providing the model with patients for whom 11 clinical inputs were paired with known outcomes, which allowed the ANN to "learn" the relevant relationships that predict outcome. The model was tested against actual outcomes in a novel subset of 100 patients derived from the same database. For comparison with traditional forms of modeling, 2 regression models were developed using the same training set and were evaluated on the same testing set. Lastly, the authors used the same 100-patient testing set to evaluate 5 neurosurgery residents and 4 neurosurgery staff physicians on their ability to predict survival on the basis of the same 11 data points that were provided to the ANN. The ANN was compared with the clinicians and the regression models in terms of accuracy, sensitivity, specificity, and discrimination. RESULTS: Compared with regression models, the ANN was more accurate (p < 0.001), more sensitive (p < 0.001), as specific (p = 0.260), and more discriminating (p < 0.001). There was no difference between the neurosurgery residents and staff physicians, and all clinicians were pooled to compare with the 5 best neural networks. The ANNs were more accurate (p < 0.0001), more sensitive (p < 0.0001), as specific (p = 0.743), and more discriminating (p < 0.0001) than the clinicians. CONCLUSIONS: When given the same limited clinical information, the ANN significantly outperformed regression models and clinicians on multiple performance measures. While this paradigm certainly does not adequately reflect a real clinical scenario, this form of modeling could ultimately serve as a useful clinical decision support tool. As the model evolves to include more complex clinical variables, the performance gap over clinicians and logistic regression models will persist or, ideally, further increase. PMID- 20020845 TI - Special issue on liver toxicology. PMID- 20020843 TI - Wrong-site craniotomy: analysis of 35 cases and systems for prevention. AB - OBJECT: The purpose of this case review was to identify and analyze existing wrong-site craniotomy (WSC) cases to determine the factors that contributed to the errors and to suggest preventative strategies for WSC. Wrong-site surgery (WSS) is a devastating surgical error that has gained increased public attention in recent years due to some high-profile cases. Despite the implementation of preventative methods such as preoperative checklists and surgical time-outs, WSS still occurs to this day. The clinical consequences of WSC are distinct compared with other types of WSS due to the unique function of the brain. METHODS: The authors searched medical, legal, and media databases and contacted state medical licensing boards to identify and gather information about WSC cases. The cases were reviewed and analyzed for factors that contributed to the errors. RESULTS: Four major categories of contributing factors were found: 1) communication breakdown; 2) inadequate preoperative checks; 3) technical factors and imaging; and 4) human error. The WSC cases are used to illustrate how these types of factors can precipitate the surgical error. Clinical outcomes and disciplinary actions are summarized. Obtaining information about the cases discovered was very challenging, in part because WSS reporting is inadequate. CONCLUSIONS: This case review demonstrates that a broad range of events and factors can cause human errors to breach patient safeguards and lead to a WSC; however, in essentially all cases the WSCs were preventable with strict adherence to comprehensive and thorough protocols. PMID- 20020846 TI - Acute Liver Toxicity Induced by 1,1-Diphenyl-2-picryl-hydrazyl (DPPH) in ICR Mice. AB - ABSTRACT The hepatotoxic effects of 1,1-diphenyl-2-picryl-hydrazyl (DPPH) in ICR mice were examined. Acute liver injury was induced by a single dose of DPPH (100 mg/kg) intraperitoneally. At 12, 24, 48, and 72 h after treatment, mice were sacrificed for serum and liver. To assess hepatotoxicity, the activities of glutamate oxaloacetate transaminases (GOT), glutamate pyruvate transaminases (GPT) in serum and glutathione peroxidase (GPx), superoxide dismutase (SOD), catalase (CAT), and the levels of malonyldialdehyde (MDA), reduced glutathione (GSH), and NO in liver were measured. The present study was also conducted so that the mice which received the DPPH at doses of 25, 50, 100, 200 mg/kg, a histopathological observation and the expression of pro-inflammatory cytokines were investigated too. The result revealed that DPPH could elevate the transaminases activities and MDA and NO levels accompanied by significant reduction in GSH level and SOD, CAT, and GPx activities. The liver from the DPPH treated mice showed prominent histopathological changes characterized by hepatocyte tumefaction and vacuolar degeneration. The mRNA levels of the pro inflammatory cytokines were elevated in the DPPH-treated groups. The above results indicated that DPPH could induce the acute liver toxicity in mice. PMID- 20020847 TI - Depletion of glutathione system enzymes of the liver and erythrocytes in a rat model of acute iron poisoning. AB - ABSTRACT The proposed mechanism of iron-induced hepatotoxicity is free radical formation. It was hypothesized that the glutathione system of the liver and erythrocytes will be affected by acute iron poisoning. Male Wistar rats, 6-8 weeks of age, were assigned to one of three groups. Group I received distilled water, group II received 400 mg/kg elemental iron, and group III received 750 mg/kg elemental iron. All groups were gavage fed. Iron concentration, glutathione, and glutathione system enzymes were then measured in the liver and erythrocytes. The hepatic level of reduced glutathione (GSH) was significantly lower in groups II (3.1 +/- 4.6 mumol/mg protein) and III (4.7 +/- 4.6 mumol/mg protein) in comparison with group I (11.5 +/- 6.2 mumol/mg protein) (p < 0.001). Hepatic levels of glutathione S-transferase (GST) were higher and glutathione peroxidase (GPX) levels were lower in group III compared to groups II and I (p < 0.001 and p < 0.001). Compared to group I, glutathione reductase (GR) was lower in groups II and III (p < 0.001). There was no correlation between GSH, oxidized glutathione (GSSG), GST, GR, and GPX levels in the erythrocytes and in the liver (p = 0.41, p = 0.48, p = 0.49, p = 0.53, p = 01.4, and p = 0.84, respectively). In conclusion, acute iron intoxication in rats is associated with depletion of reduced glutathione in the liver. PMID- 20020848 TI - Comparison of Histopathological Alterations Due to Sublethal CCl(4) on Rosy Barb (Puntius conchonius) and Amphioxus (Branchiostoma belcheri) with Implications of Liver Ontogeny. AB - ABSTRACT This study was undertaken to examine the histopathological effects of CCl(4) on rosy barbs and amphioxus with an aim to compare the homology between rosy barb liver and amphioxus digestive caecum. It was found that the 96 h LC(50) values were 23.9 +/- 4 mg/L and 18.9 +/- 2 mg/L for rosy barbs and amphioxus, respectively. Histological examinations showed that exposure to sublethal CCl(4) caused damage to the liver, kidney, and gill in rosy barb, and to the digestive caecum and gill in amphioxus. It is clear that both rosy barb liver and amphioxus digestive caecum were the prominent target organs of CCl(4), suggesting that the digestive caecum in amphioxus is homologous to the liver in rosy barb at least in respect to toxic damages of CCl(4). PMID- 20020849 TI - Interaction of sanguinarine alkaloid, isolated from argemone oil, with hepatic cytochrome p450 in rats. AB - ABSTRACT Prior studies have shown that argemone oil (AO), responsible for 'Epidemic dropsy', causes inhibition of catalytic activities of Cytochrome P450 (P450). In this study interaction of sanguinarine (SAN) alkaloid, isolated from AO, with rat hepatic P450 was investigated. Hepatic microsomes prepared from 3 methylcholantherene (3MC) treated rats when incubated with SAN (1-3 muM) resulted in a spectral peak at 385 nm and a trough at 415 nm, indicative of Type I binding. Incubation of SAN (50-200 muM) with hepatic microsomes prepared from phenobarbitone (PB) treated rats also showed a Type I spectra with a peak at 395 nm and a trough at 420 nm. Relative binding efficiency (DeltaA(max)/K(s)(app) factor) of SAN with P450 was found to be 1540 and 1030 absorbance units/nmol CYP/M for 3MC and PB induced microsomes, respectively. In a P450 spectral inhibition study SAN showed higher affinity towards 3MC eliciting inhibition at much lesser concentrations (0.25-5 muM) as compared to PB (100-300 muM). The IC50s of SAN with different catalytic markers of P450 isoforms, i.e. ethoxyresorufin-O-deethylase (EROD) for CYP1A1, was 2.8 muM and for methoxyresorufin-O-deethylase (MROD) for CYP1A2 was 2.2 muM in 3MC induced microsomes, while benzoyloxyresorufin-O-deethylase (BROD) for CYP 2B1/1A1 showed an IC50 of 50 muM but pentoxyresorufin-O-deethylase (PROD) for CYP2B1 showed no inhibition even at higher concentrations of SAN (> 60 muM) in PB-induced microsomes. These results indicate that higher affinity of SAN binding towards the CYP1A family may have a role in SAN toxicity. PMID- 20020850 TI - Epigallocatechin gallate, a green tea phytochemical, attenuates alcohol-induced hepatic protein and lipid damage. AB - ABSTRACT Oxidative damage to cellular constituents is one of the major mechanisms of alcoholic liver injury, and administration of antioxidants ameliorates alcoholic liver disease. The present study investigated the influence of (-) epigallocatechin gallate (EGCG), a major polyphenol component of green tea, on oxidant-antioxidant balance, protein, and lipid damage in liver of rats fed ethanol. Chronic ethanol administration (6 g/kg/day x 60 days) caused liver damage that was manifested by excessive formation of lipid peroxidation end products such as thiobarbituric acid-reactive substances (TBARS), lipid hydroperoxides (LHP), and conjugated dienes (CD) accompanied by a reduction in enzymic and non-enzymic antioxidant levels. Further, ethanol caused a rise in protein carbonyl formation and loss of protein thiol groups. Ethanol-fed rats exhibited increased staining for the presence of 4-hydroxynonenal (4-HNE), 3 nitrotyrosine (3-NT), and 2,4-dinitrophenol (DNP) protein adducts in liver. The detrimental effects of ethanol were alleviated upon simultaneous treatment with EGCG (100 mg/kg/day) for the last 30 days of alcohol feeding. These findings show that EGCG ameliorates protein and lipid damage induced by the hepatotoxin, ethanol. PMID- 20020851 TI - Mitigation of azathioprine-induced oxidative hepatic injury by the flavonoid quercetin in wistar rats. AB - ABSTRACT Azathioprine (AZA), one of the widely prescribed immunosuppressant drugs in organ transplantation and autoimmune diseases, could cause hepatotoxicity in the course of therapy. The current work was designed to assess the protective role of the dietary flavonoid, quercetin (QE), in oxidative hepatic damage induced by AZA. Adult male Wistar rats were divided into four treatment groups. Two groups were treated with single intraperitoneal injection of AZA (50 mg/kg body weight); one of these groups was pretreated with QE (50 mg/kg body weight) intraperitoneally once a day for 7 days. A vehicle treated control group and a QE control group were also included. Hepatotoxicity, evident from increased levels of aspartate transaminase (AST), alanine transaminase (ALT), alkaline phosphatase (ALP), lactate dehydrogenase (LDH), and gamma glutamyl transpeptidase (GGT) in serum 24 h after AZA treatment, was significantly (p < 0.05) normalized by QE pretreatment. AZA administered rats displayed declined levels of endogenous antioxidants [superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx), glutathione reductase (GR), and glutathione (GSH)], along with elevated levels of malondialdehyde (MDA). However, pretreatment with QE significantly precluded lipid peroxidation and maintained the activities of antioxidant defenses at a near normal status. Besides, AZA induced oxidative stress and subsequent DNA damage was effectively manifested by QE, which was confirmed by agarose gel electrophoresis. These findings highlight the salubrious effect of QE as a hepatoprotectant in AZA-induced oxidative stress mediated hepatic injury. PMID- 20020852 TI - Intracellular glutathione plays important roles in pyrrolizidine alkaloid clivorine-induced toxicity on L-02 hepatocytes. AB - ABSTRACT This study is aimed to evaluate the role of intracellular GSH in pyrrolizidine alkaloid clivorine-induced toxicity on human normal liver L-02 cells. Clivorine induced profound intracellular GSH depletion in a concentration dependent manner. The GSH synthesis inhibitor L-Buthionine (S, R)-sulfoximine (BSO) significantly potentiated toxicity of clivorine, while GSH and GSH synthesis precursors N-Acetyl-L-cysteine (NAC) and S-adenosyl-L-methionine (SAM) protected cells against toxicity of clivorine. Overall, the results suggest that intracellular GSH plays an important role in the toxicity of clivorine on human hepatocytes. PMID- 20020853 TI - Assessment of lung inflammation in a mouse model of smoke inhalation and burn injury: strain-specific differences. AB - ABSTRACT To test concepts developed in our ovine model of acute respiratory distress syndrome, specifically the roles of neuropeptides and other peptide mediators, a recently developed murine model of combined smoke inhalation and burn (SB) injury was extended by applying methods for quantitative assessment of acute inflammation in the lung. Mice received SB injury per protocol, n = 5 to 7 per group. Mice were anesthetized with i.p. ketamine/xylazine, endotracheally intubated, and exposed to cooled cotton smoke (4 x 30 sec for Balb/C, 2 x 30 sec for C57BL/6). After s.c. injection of 1 mL 0.9% saline, each received a 40% total body surface area (TBSA) flame burn. Buprenorphine (0.1 mg/kg) was given i.p. for postoperative analgesia; 0.9% saline was given i.p. at 4 mL/kg per %TBSA burn. Evans Blue dye (EB) was injected i.v. 15 min before sacrifice. Lung wet/dry weight ratio was measured. In other animals, after vascular perfusion with buffered saline, lungs were sampled and analyzed for myeloperoxidase (MPO), using an EIA kit, and for their content of EB dye. There was a significant (p < 0.05) increase in EB dye content, wet/dry weight ratio, and MPO 24 h after injury in Balb/C mice. Similar increases were seen in C57BL/6 mice 48 h after SB injury, but not at 24 h. C57 mice tolerated less smoke inhalation than Balb/C mice, due to postexposure apnea, and required 48 h to show significant increases in these variables. Direct comparison between animals injured by 40% TBSA burn and 2 x 30 sec smoke exposure and sacrificed after 48 h showed significantly greater abnormality in the C57BL/6 mice. The mouse model can be used effectively to assess acute inflammation in the lung. PMID- 20020854 TI - Silica Induces Plasminogen Activator Inhibitor-1 Expression through a MAPKs/AP-1 Dependent Mechanism in Human Lung Epithelial Cells. AB - ABSTRACT Plasminogen activator inhibitor-1 (PAI-1) plays an important role in the silica-induced pulmonary fibrosis. The effect of silica on the expression of PAI 1 was investigated in human lung epithelial cells (A549). Silica induced PAI-1 expression in a concentration-(50-200 mug/mL) and time-(4-24 h) dependent manner in A549 cells. Furthermore, the roles of mitogen-activated protein kinase (MAPK)/activator protein-1 (AP-1) signaling pathways in silica-induced PAI-1 expression were examined. We found that silica (200 mug/mL) treatment for 4 to 24 h resulted in AP-1 activation in A549 cells. Cells were pretreated with the AP-1 inhibitor curcumin (10, 25, 50 muM), and silica-induced PAI-1 expression was reduced by 20%, 63%, and 65%, respectively. In addition, dominant-negative mutant c-Jun (TAM67) down-regulated silica-induced PAI-1 expression by 59%. P38 kinase inhibitor SB203580 (20 muM) and Erk inhibitor PD98059 (50 muM) suppressed silica induced PAI-1 expression by 35% and 51%, respectively. Additionally, PD98059 but not SB203580 inhibited the AP-1 DNA binding activity induced by silica. The results suggest that the PAI-1 expression induced by silica may be involved in the activation of MAPKs/AP-1 signaling pathways in human lung epithelial cells. PMID- 20020855 TI - The effects of inhaled formaldehyde on oxidant and antioxidant systems of rat cerebellum during the postnatal development process. AB - ABSTRACT The aim of this study was to examine the effects of formaldehyde (FA) inhalation during the early postnatal period on some oxidant and antioxidant systems of rat cerebellum in the developmental process and to determine whether the changes were reversible or not. After birth, 0 (control), 6, or 12 ppm FA5 was given to the neonatal rats throughout 30 days. This was done by placing them for 6 h/day and 5 day/week in a glass chamber containing FA vapor. After cessation of the FA exposing process, seven rats from each group were decapitated on postnatal day (PND) 30 and the remaining seven rats were decapitated on the PND 90, and all cerebellums were removed immediately. On samples, levels of malondialdehyde (MDA) and nitric oxide (NO) and activities of total superoxide dismutase (t-SOD) and glutathione peroxidase (GSH-px) enzymes were measured. We found that activity of GSH-Px and levels of MDA and NO increased; on the other hand, activity of t-SOD decreased significantly in the rats treated with FA compared to control rats at PND 30. In general, the findings at PND 90 were similar to PND 30. Additionally, we observed that the 12-ppm FA-inhaling rats were more affected than the 6-ppm FA-inhaling rats, especially at PND 30. As a result, the present findings suggest that inhalation of FA during the early postnatal period affects the oxidant and antioxidant systems and increases some free radicals in the rat cerebellum in a dose-related manner. We think that these changes were carried on for a long time or may cause irreversible toxicity and oxidative damage. PMID- 20020856 TI - Absorption of (14)C-Cyclotrimethylenetrinitramine (RDX) from Soils through Excised Human Skin. AB - ABSTRACT Cyclotrimethylenetrinitramine (RDX), a compound used widely in bursting type munitions, is a concern for the U.S. Department of Defense because it has been detected in soil and groundwater at military installations. Dermal absorption of (14)C-RDX from acetone solutions and from two different soils was studied using excised human skin (from surgery) in flow-through diffusion cells. RDX in acetone (10 muL) or in soils (10 mg) was applied to the epidermal surface of the skin (0.64 cm(2)) and allowed to transverse the skin and become dissolved in a reservoir of receptor fluid that was maintained in contact with the dermal surface. The reservoir was of the flow-through type and receptor fluid was pumped at a rate of 1.5 mL/h. Receptor fluid was collected every 6 h for 24 h. Because the bioavailability of a chemical from soils depends on soil composition, dermal absorption of (14)C-RDX from both a low-carbon (1.9%) and a high-carbon (9.5%) soil was assessed. At the conclusion of the experiment, the RDX remaining on the skin was washed with soap and water using cotton swabs, and the radioactivity present in washings was determined. The stratum corneum was removed from the deeper epidermis and radioactivity found in that layer was not considered in calculations of dermal absorption. The dermal absorption of RDX was relatively low. Only about 5.7 +/- 1.9% of the RDX that had been applied in acetone was found in the skin (epidermis and dermis) (3.2 +/- 1.9) and receptor fluid (2.5 +/ 1.8) combined (over the full 24-h duration of the study). The levels of RDX found in the skin layers were stratum corneum 2.1%, epidermis 0.83%, and dermis 0.45%. The total recovery of applied dose (receptor fluid, skin, and washings) was about 80%. The extent of RDX absorption from soil was even lower than from acetone. Approximately 2.6 +/- 1.1% of the RDX applied in the low-carbon soil and 1.4 +/- 0.41% applied in the high-carbon soil was found in receptor fluid and skin in 24 h. The total recovery of the applied dose (receptor fluid, skin, and washings) was about 87% for the low-carbon soil and 94% for the high-carbon soil. Thus, the dermal absorption of RDX from soils was reduced considerably when compared with absorption from acetone and absorption was lower in the high-carbon soil than in the low-carbon soil. PMID- 20020857 TI - Effect of n6-cyclopentyladenosine on ouabain-induced toxicity in isolated Guinea pig atria. AB - ABSTRACT The effect of N6-cyclopentyladenosine (CPA), an A(1)-selective adenosine agonist, was studied on ouabain-induced toxicity in spontaneously beating isolated guinea pig atria. CPA (2-16 nM) produced a dose-dependent decrease in the force of contractions (34%-51%) and in the rate of contractions (22%-48%). CPA significantly increased the time of onset of arrhythmia (toxicity) induced by ouabain (1.2 muM) when it was administered 10 min before ouabain was added in organ bath. Ouabain (1.2 muM) alone produced arrhythmia at 7 min and either asystole or standstill at 22 min. CPA (8 nM) increased the time required to produce arrhythmia to 27.5 min and prolonged beating atria to more than 63 min and prevented the occurrence of asystole. These findings indicated that CPA produces direct cardiac action, probably due the inhibition of cardiac Na(+) and Ca(2+) channels. Moreover, our results suggest that CPA may reduce the membrane conduction through inhibition of ionic channels, which decrease ouabain-induced toxicity. PMID- 20020858 TI - On the mechanisms underlying poisoning-induced rhabdomyolysis and acute renal failure. AB - ABSTRACT The clinical syndrome of rhabdomyolysis is caused by injury of skeletal muscles resulting in release of intracellular muscle constituents. Drug poisoning is one of the causes of severe rhabdomyolysis. Severe electrolyte disorders and acute renal failure may occur in rhabdomyolysis, leading to life-threatening situations. Early initiation of renal replacement therapy can help improve outcome. In the present retrospective study, medical records of 181 patients suspected of rhabdomyolysis from Loghman-Hakim Hospital in the period of 2004 to 2005 were reviewed. A creatinine phosphokinase (CPK) value of greater than five times normal (>/=975 IU/L) was the basis for confirmation of a rhabdomyolysis diagnosis. An increased serum creatinine level of more than 30% was the basis for acute renal failure diagnosis. Out of 156 patients, 100 were male with an age range of 13 to 78 years. One hundred and two (92%) patients had CPK >975 U/L, and 36 patients (28.6%) had a 30% or more increase in their creatinine level during their admission days. Mean fluid intake was the same in patients with renal failure and those without renal failure. In 8.3% of the cases, multiple drug poisoning was observed. The most common compound overdose associated with rhabdomyolysis was opium. It is concluded that fluid therapy alone is not adequate in the management of acute renal failure in rhabdomyolysis. Therefore, other etiological factors are involved that remain to be elucidated by further studies. PMID- 20020859 TI - Biochemical Evaluation of Human DNA-Lysine Photoadduct Treated with Peroxynitrite. AB - ABSTRACT Peroxynitrite is a reactive oxidant produced from nitric oxide ((.)NO) and superoxide anion (O(2)(.-)). It is produced by the body in response to environmental toxins, stress, ultraviolet light, ischemia/reperfusion, inflammation, etc. In vivo, peroxynitrite is formed in macrophages, endothelial cells, platelets, leukocytes, and neurons. It reacts with a variety of biomolecules including proteins, lipids, and DNA. We have investigated the photochemical addition of lysine to native DNA in view of its potential importance in the photo-cross-linking of histones to DNA in chromatin. Lysine-and arginine-rich histone H1 in nucleosome on modification by physical, chemical, or environmental agents forms histone-DNA adducts. We have characterized the photoadducts by absorption, fluorescence, and chromatographic methods. The UV absorption spectra of the DNA-lysine photoadduct showed hyperchromism, indicating structural distortions in DNA either due to single-strand breaks or opening of the double helix at the site of lysine conjugation. On peroxynitrite treatment, the melting temperature (T(m)) of the DNA-lysine adduct increased by 15 degrees C compared to the native DNA-lysine adduct. A decrease in the fluorescence intensity of the DNA-lysine photoadduct with respect to the modified adduct was observed. The gel filtration profile of the peroxynitrite-modified adduct was also found to be different from that of the native DNA and DNA-lysine photoadduct. Hence, the peroxynitrite-modified photoadduct may have important implications in toxicology, mutagenesis, and carcinogenesis. PMID- 20020860 TI - Impact of dietary restriction on peroxidative effects of nickel chloride in wistar rats. AB - ABSTRACT The purpose of this study, carried out in Wistar rats, was to evaluate the protective effect of dietary restriction (performed by intermittent fasting) against oxidative stress induced by a low concentration of nickel chloride in kidney, liver, uterus, and ovary. Lipid peroxidation (TBARS), catalase activity, and the levels of vitamins E and A in the blood were investigated in rats feed for 1 month either daily (N) or 1 day over two (intermittent fasting, IF) and then injected (NNi, IFNi) or not with nickel chloride (30 mumoles/kg body weight/day) for 10 days. Ni induced a significant increase of TBARS in organs of N rats. Intermittent fasting alone or associated to nickel treatment did not result in TBARS change in IF and IFNi rats. Catalase activity levels were found to be similar in N and IF rats. In Ni-treated rats a transient increase of catalase activity appeared at day 1 in the kidney and days 1 and 3 in the liver. Then, catalase activity was found to be inhibited until day 10. In the uterus and ovary, catalase activity was always found to be inhibited. In IFNi rats, no significant increase of catalase activity was observed as compared to IF rats. Vitamin E was inhibited from the 1st to the 10th day in Ni rats, whereas no significant changes were noted in IFNi rats. A moderate decrease of vitamin A was only found at days 1 and 3 in Ni rats. In conclusion, intermittent fasting is able to protect from oxidative stress induced by low concentration of Ni, but catalase and Vitamins E and A do not seem to be involved. PMID- 20020861 TI - Identification of Genes Responding to Low-Dose Arsenite Using HiCEP. AB - ABSTRACT Chronic ingestion of arsenic in polluted food and water can cause various human disorders including skin and lung cancers. Sensitive biomarkers from human tissue/cells could help to prevent chronic intoxication with low-dose arsenite. Using High-Coverage Expression Profiling (HiCEP), an Amplified Fragment Length Polymorphism (AFLP)-based gene expression profiling technique, we analyzed the expression of approximately 11,000 genes in human lung fibroblasts (HFLIII) and compared the profiles between cells, treated and untreated with 1 muM sodium arsenite (NaAsO(2)). Hundreds of genes appeared upregulated and downregulated more than two-fold, 2 h after the treatment. Marked induction was found (>4.4 fold) in a few genes including HMOX1, INHBA, and ANKRD11. Induction of the HMOX1 was detected with a dose of arsenite at as low as 0.3 muM (0.04 ppm) and reached its maximum at 4 h after the treatment. The arsenite-induced HMOX1 expression was attenuated by the promoted glutathione (GSH) synthesis by N-acetyl-L-cysteine (NAC). However, it was not affected by pretreating the cells with general radical scavengers, consistent with the fact that ionizing radiation at either high-or low-doses has never induced HMOX1 in the same assay system. Thus, induction of HMOX1 gene is highly sensitive and also selective against arsenite in the cells. The present process could provide a useful strategy for exploring biomarkers that might help in assessing the known and unknown risks of any natural and artificial toxic reagents. PMID- 20020863 TI - Testicular toxicity and mechanisms of chlorotoluron compounds in the mouse. AB - ABSTRACT Background: Previous studies have reported that chlorotoluron has adverse effects on male mice. Few studies, however, have investigated the mechanism of action of these herbicides. The effects of chlorotoluron and atrazine ingestion on male mouse reproductive function during the peripubertal period are reported here. METHODS: A total of 16 experimental groups and one control group were used in this study. Each experimental group was treated with chlorotoluron or atrazine alone or in combination in different doses. After mice were sacrificed, superoxide dismutase activity in blood was measured, weight ratios of testis to body were calculated, and testis tissues were examined by light and transmission electron microscopy. RESULTS AND CONCLUSIONS: The decreased weight ratio of mouse testis to body weight resulted in a reduction in the relative testis weight. Experiments on SOD enzyme activity in vivo showed that the synergistic effect of the two compounds was more notable at higher concentrations. Histological analyses of testes of animals treated with herbicides showed that the epithelium of seminiferous tubules was loosely arrayed and lacked order, spermatogenic cells were shed, and fewer layers formed. By electron microscopy it was noted that mitochondria in the seminiferous epithelium appeared vacuolated; nuclei were enlarged and irregular. In addition, the number of Sertoli cells was reduced, and part of the tight junction was destroyed. The results demonstrated that these herbicides induced degeneration and severe pathological changes in a dose-dependent manner. The toxic effects of chlorotoluron were more potent than those of atrazine when each was administrated alone. Moreover, when administered in combination, the two compounds had synergistic toxic effects. PMID- 20020862 TI - Metabolomics in lung inflammation:a high-resolution (1)h NMR study of mice exposedto silica dust. AB - ABSTRACT Here we report the first (1)H NMR metabolomics studies on excised lungs and bronchoalveolar lavage fluid (BALF) from mice exposed to crystalline silica. High-resolution (1)H NMR metabolic profiling on intact excised lungs was performed using slow magic angle sample spinning (slow-MAS) (1)H PASS (phase altered spinning sidebands) at a sample spinning rate of 80 Hz. Metabolic profiling on BALF was completed using fast magic angle spinning at 2 kHz. Major findings are that the relative concentrations of choline, phosphocholine (PC), and glycerophosphocholine (GPC) were statistically significantly increased in silica-exposed mice compared to sham controls, indicating an altered membrane choline phospholipids metabolism (MCPM). The relative concentrations of glycogen/glucose, lactate, and creatine were also statistically significantly increased in mice exposed to silica dust, suggesting that cellular energy pathways were affected by silica dust. Elevated levels of glycine, lysine, glutamate, proline, and 4-hydroxyproline were also increased in exposed mice, suggesting the activation of a collagen pathway. Furthermore, metabolic profiles in mice exposed to silica dust were found to be spatially heterogeneous, consistent with regional inflammation revealed by in vivo magnetic resonance imaging (MRI). PMID- 20020864 TI - Quantification of phenol, phenyl glucuronide, and phenyl sulfate in blood of unanesthetized rainbow trout by online microdialysis sampling. AB - ABSTRACT Free concentrations of phenol (PH), phenyl glucuronide (PG), and phenyl sulfate (PS) were measured in the bloodstream of unanesthetized rainbow trout by online microdialysis (MD) sampling. Preliminary studies were conducted to optimize the MD system and evaluate three retrodialysis calibration standards: p nitrophenyl glucuronide (PNPG), p-nitrophenyl sulfate (PNPS), and [(14)C]-phenol ((14)C-PH). PG and PNPG exhibited nearly identical dialyzing properties in vitro (saline and plasma) and in vivo (muscle tissue and dorsal aorta). A similar result was obtained for PS and PNPS. In vivo studies were therefore performed using PNPG, PNPS, and (14)C-PH as retrodialysis calibrators for PG, PS, and PH, respectively. The utility of MD sampling for kinetic studies with fish was investigated by implanting MD probes into the dorsal aorta of spinally transected rainbow trout. Each fish was then exposed to PH in water in a respirometer metabolism chamber. The free concentration of PH in blood reached a steady-state level within 12 h of initiating the exposure. A steady state for PS was generally established within 24 h, while free concentrations of PG tended to increase throughout the exposure. Terminal plasma samples were dialyzed using the same probe employed in each experiment. Analyte concentrations determined in this manner were in good agreement with calculated in vivo values. The methods described in this study can be used to collect kinetic data sets of high temporal resolution while eliminating artifacts often associated with conventional blood sampling methods. PMID- 20020865 TI - DPPH Radical Scavenging Activity of Several Naturally Occurring Coumarins and Their Synthesized Analogs Measured by the SIA Method. AB - ABSTRACT Coumarins, naturally occurring compounds derived from benzopyran, have recently been studied extensively for their antioxidant properties. A lot of coumarins have been isolated and identified from natural sources and many others have been synthesized. It is also known that pharmacological and biochemical properties and thus also therapeutic application of simple coumarins depend upon the pattern of their substitution. As a part of studies of biological effects, four naturally occurring coumarins and 18 synthesized analogs of several compounds were assayed for 2,2-diphenyl-1-picrylhydrazyl radical (DPPH) scavenging activity. For this purpose the highly reliable DPPH test modified to be performed by sequential injection analysis (SIA) system was used. This in our laboratory-developed method was originally proposed for antioxidant screening of large series of plant extracts. In this assay, the DPPH test using the SIA method was used for fast and sensitive evaluation of EC(50) of coumarins. The evaluation of EC(50) of a single compound takes only 15 to 30 min. The structure-activity relationships of tested compounds are also established. The results verified 7,8 dihydroxy-4-methylcoumarins as excellent DPPH radical scavengers. Obtained results correspond with those of other studies and suggest the SIA procedure as a suitable method for fast and sensitive antioxidant analysis of various types of compounds. PMID- 20020866 TI - N-(2-Hydroxylpropyl)-Methacrylamide-Attached Doxorubicin Induces Cytotoxicity to Prostate Cancer Cell Line DU145. AB - ABSTRACT Successful use of doxorubicin as an antitumor agent is limited by its cardiotoxicity, which takes different forms and results from multiple biochemical alterations in the cell. This study addresses the possibility to overcome these adverse effects by studying the effects of N-(2-hydroxypropyl)methacrylamide (HPMA) polymer, which contains doxorubicin attaching to it. Both time-and dose dependent studies were conducted using DU145 cell lines. The results of this study reveal that doxorubicin attached to HPMA can be used successfully in treating cancer instead of doxorubicin alone, which may open a new gate for clinicians and scientists, leading to overcoming the resistance and side effects of the currently used antitumor agents. PMID- 20020867 TI - (-) Epigallocatechin Gallate (EGCG) Prevents Lipid Changes and Collagen Abnormalities in Chronic Ethanol-Fed Rats. AB - ABSTRACT The objective of the study is to examine the influence of (-) epigallocatechin gallate (EGCG), a green tea component, on lipid and collagen abnormalities in chronic ethanol-fed rats. Solubility properties, aldehyde content, fluorescence, and peroxidation were analyzed in collagen samples isolated from liver. Chronic alcoholism (6 g/kg/day x 60 days) was associated with fatty liver and collagen accumulation. Significant alterations in the levels of lipids (cholesterol, phospholipids, free fatty acids, and triglycerides) and total collagen were observed in liver. Collagen obtained from ethanol-fed rats showed alterations in solubility properties, increased fluorescence, peroxidation, and aldehyde content. Coadministration of EGCG along with ethanol significantly reduced the levels of liver lipids and collagen, improved the solubility properties of collagen, and caused a reduction in cross-linking as evidenced by a decrease in fluorescence, peroxidation, and aldehyde content. Histology of liver sections of ethanol-fed rats showed accumulation of fat and collagen, which were largely prevented by EGCG administration. The possible mechanisms in the protective action of EGCG in alcoholic liver disease are suggested and discussed. PMID- 20020868 TI - Safety of a Novel Calcium/Potassium Salt of Hydroxycitric Acid (HCA-SX): I. Two Generation Reproduction Toxicity Study. AB - ABSTRACT (-)-Hydroxycitric acid (HCA), a natural plant extract from the dried fruit rind of Garcinia cambogia, has been reported to inhibit fat synthesis and reduce food intake. The objective of this study was to evaluate the effects of a novel calcium/potassium salt of (-)-hydroxycitric acid (HCA-SX) on the reproductive systems of male and female rats, the postnatal maturation and reproductive capacity of their offspring, and possible cumulative effects through multiple generations. Sprague-Dawley rats (30/sex/group) were maintained on feed containing HCA-SX at dose levels of 0, 1000, 3000, or 10,000 ppm for 10 weeks prior to mating, during mating, and, for females, through gestation and lactation, across two generations. During the period of study, animals were examined daily for signs of clinical toxicity and their body weight and feed consumption were recorded twice a week. For the parents (F(0) and F(1)) and the offspring (F(1) and F(2a)), reproductive parameters such as fertility and mating, gestation, parturition, litters, lactation, sexual maturity, and development of offspring were assessed. At termination, necropsy and histopathological examinations were performed on all animals. Dietary exposure of HCA-SX to parental male and female rats of both (F(0) and F(1)) the generations during the premating and mating periods, for both sexes, and during gestation and lactation in case of female rats, did not reveal any remarkable incidence of mortality or abnormal clinical signs. Compared to respective controls, HCA-SX exposure did not affect feed consumption or body weight at any of the exposure levels. HCA-SX exposure did not affect reproductive performance as evaluated by sexual maturity, fertility and mating, gestation, parturition, litter properties, lactation, and development of the offspring. Based on the results of this study, the parental as well as the offspring no-observed-adverse-effect level for HCA-SX was determined to be greater than 10,000 ppm in diet or equivalent to 1018 and 1524 mg/kg body weight/day in male and female rats, respectively. PMID- 20020869 TI - Safety of a Novel Calcium/Potassium Saltof (-)-Hydroxycitric Acid (HCA-SX): II.Developmental Toxicity Study in Rats. AB - ABSTRACT (-)-Hydroxycitric acid (HCA), active constituent (10%-30%) of the dried fruit rind of Garcinia cambogia, is commonly used as a dietary supplement for weight management. The objective of the present study was to evaluate the teratogenic potential of a novel calcium/potassium salt of HCA (HCA-SX) in Sprague-Dawley rats. Due to its potential to affect fat synthesis and reduce food intake, processes that are often crucial in normal fetal development, this teratology study was undertaken as part of a multigeneration reproductive investigation. The animals in this study were selected randomly after weaning from each F(2b) litter of the F(1) generation from the two-generation reproductive toxicity study. To start the teratology study, Sprague-Dawley rat pups ( approximately 30/sex/group) from the F(2b) generation were allowed to grow up to 10 to 12 weeks of age before mating. The rats in the treatment group were exposed directly to HCA-SX through feed, while prior to their weaning, they had indirect exposure to the test material during lactation. The dietary exposure levels were the same as those employed for the two-generation reproductive toxicity study, viz. 1000, 3000, or 10,000 ppm. Following mating at maturity, the pregnant rats were observed daily for clinical signs of adverse effects, and body weight and feed consumption were recorded. On day 20 of gestation, animals were subjected to a necropsy and cesarean section to examine the uterus, ovaries, and fetuses for assessment of different parameters of pregnancy and embryo-fetal defects. Despite a slight (13%) lowering of maternal body weight gain during gestation period in the group receiving 10,000 ppm HCA-SX, no evidence of maternal toxicity, adverse effects on the parameters evaluated for the gravid uteri, external abnormalities in the fetuses, soft tissue abnormalities in the fetuses, or skeletal abnormalities in the fetuses were noted. Based on the results of this developmental toxicity study, conducted in continuation of a two generation reproductive toxicity study, HCA-SX was not found to be teratogenic in the Sprague-Dawley rat at the dietary exposure levels of 1000, 3000, and 10,000 ppm, equivalent to the dose levels of 103, 352, or 1240 mg/kg/day, respectively. PMID- 20020870 TI - Methods for detecting chemical-chemical interaction in toxicology. AB - ABSTRACT When two toxicants overtly producing similar effects are given together, the effect of the combination may either be equal to the algebraic sum of their individual effects or may be more than or less than the algebraic sum of their individual effects. Therefore, to ascertain the nature of interaction precisely and accurately, whether it is additive, supra-additive (synergistic), or infra additive (antagonistic), appropriate experimental designs are required to be followed. Factorial experiments requiring large numbers of animals are not only uneconomical, but also fail to reveal the exact nature of interaction with regard to addition, synergism, or antagonism. Scientists have therefore devised several simple experimental designs that are based on the dose-response curves of individual interacting substances and the dose-response curve of fixed-proportion combinations of these toxic substances. Among the reported experimental designs, the most efficient methods include the isobolographic method and the interaction index method. The present review aims to discuss the principle and methodology behind the experimental designs used to study interaction of binary toxic mixtures, to make it comprehensible for researchers engaged in the field of toxicology. A procedure by which nature of interaction can be ascertained with the help of the interaction index using Student t-test has also been explained. The review has been supported with appropriate illustrations. PMID- 20020871 TI - PGE from Octopus aegina Induces Apoptosis in Ehrlich's Ascites Carcinoma of Mice. AB - ABSTRACT The present study was carried out to assess the antitumor effect of venomous peptide from the cephalopod Octopus aegina on Ehrlich's ascites carcinoma (EAC). Male albino Swiss mice were used in the present study. Four groups of animals were treated with three doses of the sublethal dose of venom, 15, 75, and 150 mug/kg body weight (intraperitoneal injection), along with the standard drug 5-fluorouracil (20 mg/kg b.w.). After 10 days of treatment, six animals from each group were sacrificed for the biochemical analysis and the rest were left to calculate the mean survival time. In EAC-bearing mice, mean lifespan, tumor volume, hemoglobin, red blood cells, and lymphocytes were significantly decreased when compared to the normal animals. While body weight, neutrophils, and viable tumor cell count were increased in the EAC-bearing mice, these changes were brought back to near normal levels in different treatment groups. The macromolecule concentration of peritoneal cells, such as DNA, RNA, and protein, were altered in the EAC-bearing mice and observed to be near normal in the treatment groups. The caspase-3 activity was significantly increased in the peritoneal cells of the treatment groups when compared to the EAC-bearing mice. The role of apoptotic cascade in EAC cell death was confirmed by the DNA fragmentation on agarose gel. Apart from the antitumor effect, octopus venom reduced the tumor burden on the liver and altered the changes in the activities of alanine transaminase (ALT), aspartate transaminase (AST), and alkaline phosphatase (ALP). Therefore, the venom from O. aegina has a potential antitumor effect on the EAC-bearing mice. PMID- 20020872 TI - Effect of repeated low-dose organophosphorothionate pesticide exposure on digoxin pharmacokinetics in rats; a possible interaction involving p-glycoprotein. AB - ABSTRACT Aside from acute occupational exposure, an important part of the population may be chronically exposed to the trace amounts of organophosphorothionate pesticides (OPTs) via residues in nutrients and drinking water. P-glycoprotein (P-gp) is a transmembrane protein responsible for the efflux of numerous drugs. OPTs were shown to inhibit P-gp function in vitro and increase its expression in vivo. Digoxin is a probe drug for the investigation of P-gp. To evaluate the effect of repeated low-dose OPT exposure on P-gp, commercial formulations of diluted OPT or tap water were administered to female Wistar rats for 8 consecutive days. On the ninth day each group was further divided into two groups and digoxin was administered either intraduodenally (ID) or intravenously (IV). Blood sampling and bile and urine collection were taken during 6 h at various intervals. The peak concentration in serum (C(max)) of digoxin was found to be decreased and the mean absorption time (MAT) was significantly increased in the digoxin OPT group. The mean residence time was significantly elevated in the digoxin(ID) OPT group. The biliary excretion% digoxin was significantly increased in the digoxin OPT group, while the renal excretion% digoxin rose only in the digoxin(ID) OPT group. No significant differences in time to reach C(max) (t(max)), area under the plasma concentration time curve (AUC)(0-360), area under the moment curve (AUMC)(0-360), and bioavailability (F) were detected. In our study, repeated low-dose OPT exposure reduced the absorption and increased the excretion% digoxin, which may be related to enhanced P-gp expression. However, alterations of digoxin pharmacokinetic parameters did not change the systemic availability of digoxin. PMID- 20020873 TI - Modulatory effects of black tea polyphenols on rat forestomach carcinogenesis. AB - ABSTRACT The present study was designed to evaluate the chemopreventive effects of black tea polyphenols (Polyphenon-B) on N-methyl-N'-nitro-N-nitrosoguanidine (MNNG)-induced gastric carcinogenesis in Wistar rats. Intragastric administration of MNNG induced well-differentiated squamous cell carcinomas that showed diminished mitochondrial lipid and protein oxidation and an increase in antioxidants. In contrast to tumor tissue, the liver mitochondria of tumor bearing animals showed elevated lipid and protein oxidation with compromised antioxidant defenses. Dietary administration of Polyphenon-B effectively suppressed MNNG-induced stomach tumors, modulated mitochondrial lipid and protein oxidation, and enhanced antioxidant enzyme activities in the stomach and liver. Our results suggest that Polyphenon-B may exert its chemopreventive effects by modulating mitochondrial cellular redox status in the tumor as well as in the host liver. PMID- 20020874 TI - Contact sensitizer potassium dichromate alters lymphocyte populations in draining lymph nodes and blood in mice. AB - ABSTRACT Chromium is a common human contact allergen, but it is not known whether chromates cause contact hypersensitivity by immunological mechanisms similar to those induced by strong haptens. To understand the immunological events of contact hypersensitivity to chromates, we investigated whether and how chromate sensitization alters lymphocyte subsets in draining lymph nodes (DLNs), blood, and spleens in mice. BALB/c mice were sensitized by painting their ears with 0.5% potassium dichromate or vehicle alone on 3 consecutive days. Flow cytometric analysis of lymphocyte surface antigens showed that the chromate exposure significantly increased the percentage of B cells and decreased the percentages of T cells in the DLNs. This was accompanied by a relative increase in T cells and a relative decrease in B cells in peripheral blood. In contrast to the chromate, sodium dodecyl sulfate (a skin irritant) did not affect B cells or T cells in the three compartments. Moreover, sensitization to the chromate led to dose-dependent decreases in the percentages of CD4(+) T cells and CD8(+) T cells in the DLNs. However, CD4(+) and CD8(+) memory T cells were significantly increased in the blood and DLNs of the chromate-sensitized mice. Additionally, the percentage of B cells in the DLNs but not blood was dose-dependently increased in the chromate-sensitized mice. Histologically, B-cell areas were dramatically enlarged in the DLNs of the chromate-sensitized mice. Thus, this report provides basic information to further elucidate the role of individual lymphocyte subsets in contact hypersensitivity to chromates. PMID- 20020875 TI - Testicular toxicity and mechanisms of chlorotoluron compounds in the mouse. AB - ABSTRACT Previous studies have reported that chlorotoluron has adverse effects on male mice. Few studies, however, have investigated the mechanism of the action of these herbicides. The study of the effects of chlorotoluron and atrazine ingestion on male mice reproductive function during a critical developmental period, the peripubertal period, is reported here. Chlorotoluron and/or atrazine were administered by PO gavage to mice from day 27 to day 52. The decreased weight ratio of mouse testis to body weight resulted in a reduction in the relative testis weight. Histological study of testis showed that the seminiferous epithelium arrayed loosely and disorderedly, spermatogenic cells became shed, and fewer layers were formed. With the use of an electron microscope, it was noted that mitochondria in the seminiferous epithelium appeared vacuolated; karyotheca swelled and bent. In addition, the number of Sertoli cells was reduced, and part of the tight junction was destroyed. The results of the present study demonstrated that herbicides in all administrated doses caused detrimental changes in mice. The toxic effects of chlorotoluron were more potent than those of atrazine when administrated alone. Moreover, the two compounds have synergistic toxic effects with combination use. All dose paradigms induced degeneration and severe pathological changes in a manner of dose response. PMID- 20020876 TI - How Sublethal Fenitrothion is Toxic in Carp (Cyprinus carpio L.) Fingerlings. AB - ABSTRACT The purpose of this study was to evaluate the effects of fenitrothion, an organophosphothionate insecticide, on selected serum hematological and biochemical parameters of carp (Cyprinus carpio L.) fingerlings and investigate histopathological changes after acute dose of exposure. Among those, cortisol and glucose are evaluated as responses of early life stages of carp to acute stress and hematological blood parameters are evaluated to investigate the mechanism of toxicity via histopathological changes to the standard test organism. No significant histological findings were observed in carp tissues (gills, liver, kidneys, spleen, gonads, brain, muscle, and skin) exposed to a sublethal concentration of 10 mg/L fenitrothion and controls. Hematocrit and erythrocyte counts were decreased significantly by exposure to fenitrothion when compared to the control group (25.26% +/- 0.82%, 1455 +/- 90.3 [10(3)/muL] vs. 28.83% +/- 1.60%, 1865 +/- 74.5 [10(3)/muL], P < 0.05), respectively. Leucocyte and thrombocyte counts did not change. Increases in plasma cortisol levels were found in the experimental group compared to controls (34.2 +/- 1.9 vs. 18.0 +/- 2.2, P < 0.05) respectively, while glucose, chloride, sodium, potassium, phosphorous, and brain MDA and FOX levels did not show significant differences from those of control (P > 0.05). As a result, we can conclude that early life stages of fish are very susceptible to the adverse effects of toxicants. PMID- 20020877 TI - Mammalian iron metabolism. AB - ABSTRACT Iron is an essential transition metal for mammalian cellular and tissue viability. It is critical to supplying oxygen through heme, the mitochondrial respiratory chain, and enzymes such as ribonucleotide reductase. Mammalian organisms have evolved with the means of regulating the metabolism of iron, because if left unregulated, the resulting excess amounts of iron may induce chronic toxicities affecting multiple organ systems. Several homeostatic mechanisms exist to control the amount of intestinal dietary iron uptake, cellular iron uptake, distribution, and export. Within these processes, numerous molecular participants have been identified because of advancements in basic cell biology and efforts in disease-based research of iron storage abnormalities. For example, dietary iron uptake across the intestinal duodenal mucosa is mediated by an intramembrane divalent metal transporter 1 (DMT1), and cellular iron efflux involves ferroportin, the only known iron exporter. In addition to duodenal enterocytes, ferroportin is present in other cell types, and exports iron into plasma. Ferroportin was recently discovered to be regulated by the expression of the circulating hormone hepcidin, a small peptide synthesized in hepatocytes. These recent studies on the role of hepcidin in the regulation of dietary, cellular, and extracellular iron have led to a better understanding of the pathways by which iron balance in humans is influenced, especially its involvement in human genetic diseases of iron overload. Other important molecular pathways include iron binding to transferrin in the bloodstream for cellular delivery through the plasma membrane transferrin receptor (TfR1). In the cytosol, iron regulatory proteins 1 and 2 (IRP1 and IRP2) play a prominent role in sensing the presence of iron in order to posttranscriptionally regulate the expression of TfR1 and ferritin, two important participants in iron metabolism. From a toxicological standpoint, posttranscriptional regulation of these genes aids in the sequestration, control, and hence prevention of cytotoxic effects from free floating nontransferrin-bound iron. Given the importance of dietary iron in normal physiology, its potential to induce chronic toxicity, and recent discoveries in the regulation of human iron metabolism by hepcidin, this review will address the regulatory mechanisms of normal iron metabolism in mammals with emphasis on dietary exposure. It is the goal of this review that this information may provide in a concise format our current understanding of major pathways and mechanisms involved in mammalian iron metabolism, which is a basis for control of iron toxicity. Such a discussion is intended to facilitate the identification of deficiencies so that future metabolic or toxicological studies may be appropriately focused. A better knowledge of iron metabolism from normal to pathophysiological conditions will ultimately broaden the spectrum of the usefulness of this information in biomedical and toxicological sciences for improving and protecting human health. PMID- 20020878 TI - Renal Lysosomal Functions on Exposure to Scatophagus argus Venom in Experimental Mice. AB - ABSTRACT Scatophagus argus venom was extracted by homogenization and centrifugation. The lysosome-enriched fraction of mice kidney was isolated by differential centrifugation. To study the effect of S. argus venom on lysosomal membrane stability, acid phosphatase and cathepsin D, the two marker enzymes for lysosome, were assayed. In the renal lysosomal enriched fraction incubated with different concentrations of venom and at various time intervals, in vivo and in vitro studies were done. Studies reveal that with increasing concentration of venom and time, the activity of the enzymes increased, which showed that the membrane integrities decreased for both in vitro and in vivo studies. The activity of the enzymes acid phosphatase and cathepsin D showed a dose-dependent relationship with S. argus venom. PMID- 20020879 TI - Beneficial effect of caffeic Acid on alcohol-induced alterations in lipid peroxidation and antioxidant defense in rats. AB - ABSTRACT We studied the effect of caffeic acid on alcohol-induced oxidative stress in alcohol-induced toxicity in rats. The elevation in the levels of lipid peroxidation in alcohol administration was accompanied by a significant decrease in the levels of vitamins C and E, superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx), reduced glutathione (GSH), and glutathione S transferase (GST) in the liver and kidney. Oral administration of caffeic acid at a dose of 12 mg/kg/body weight for 45 days significantly reduced the severe oxidative stress in alcohol toxicity as evidenced by the decrease in the levels of lipid peroxidation with a simultaneous increase in the level of enzymic and nonenzymic antioxidants in liver and kidney. Histological studies in the liver and kidney also showed that administration of caffeic acid to alcohol-treated rats resulted in a marked reduction in alcohol-induced pathological changes. These results suggest that administration of caffeic acid to alcohol-supplemented rats alleviates the alcohol-induced oxidative damage in the liver and kidney. PMID- 20020880 TI - The evaluation of altered antioxidative defense mechanism and acetylcholinesterase activity in rat brain exposed to chlorpyrifos, deltamethrin, and their combination. AB - ABSTRACT The present investigation was undertaken to examine the effects of chlorpyrifos (CP, an organophosphate) and deltamethrin (DM, a pyrethroid pesticide) treatments at low and high doses and their combination on lipid peroxidation and antioxidant enzyme activities such as SOD, GSH-Px, and CAT and also AChE activity in rat brain following 16 weeks of exposure. Antioxidative defense mechanisms and lipid peroxidation in rat brain tissues display different responses depending on different pesticide treatments and doses. Biochemical analysis showed that administration of chlorpyrifos and deltamethrin causes brain damage via production of MDA and inhibition of AChE. In the present study, we observed that LPO levels are higher at low doses than at high doses, but DM is more pronounced than CP in this increase. Observations show that SOD increases with the dose, CAT decreases at the higher dose, and GSH-Px again increases with the dose in both pesticide treatments, but especially in DM. Experimentally, oxidative-antioxidative balance is more affected by deltamethrin treatment than that of chlorpyrifos. The effects of combination of DM and CP on LPO may be due to functional, dispositional, or chemical antagonism, while the effect of the combination on CAT activity may be synergistic. PMID- 20020881 TI - Mercury activates phospholipase a(2) and induces formation of arachidonic Acid metabolites in vascular endothelial cells. AB - ABSTRACT Currently, mercury has been identified as a risk factor in cardiovascular diseases among humans. Here, we tested our hypothesis that mercury modulates the activity of the vascular endothelial cell (EC) lipid signaling enzyme phospholipase A(2) (PLA(2)), which is an important player in the EC barrier functions. Monolayers of bovine pulmonary artery ECs (BPAECs) in culture, following labeling of membrane phospholipids with [(3)H]arachidonic acid (AA), were exposed to the inorganic form of mercury, mercury chloride, and the release of free AA (index of PLA(2) activity) and formation of AA metabolites were determined by liquid scintillation counting and enzyme immunoassay, respectively. Mercury chloride significantly activated PLA(2) in BPAECs in a dose-dependent (0 to 50 muM) and time-dependent (0 to 120 min) fashion. Metal chelators significantly attenuated mercury-induced PLA(2) activation, suggesting that cellular mercury-ligand interaction is required for the enzyme activation and that chelators are suitable blockers for mercury-induced PLA(2) activation in ECs. Sulfhydryl (thiol-protective) agents, calcium chelating agents, and cPLA(2) specific inhibitor also significantly attenuated the mercury-induced PLA(2), suggesting the role of thiol and calcium in the activation of cPLA(2) in BPAECs. Significant formation of AA metabolites, including the release of total prostaglandins, thromboxane B(2), and 8-isoprostane, were observed in BPAECs following their exposure to mercury chloride. Mercury chloride induced cytotoxicity as observed by the altered cell morphology and enhanced trypan blue uptake, which was attenuated by the cPLA(2) inhibitor AACOCF(3). The results of this study revealed that inorganic mercury-induced PLA(2) activation through the thiol and calcium signaling and the formation of bioactive AA metabolites further demonstrated the association of PLA(2) with the cytotoxicity of mercury in ECs. Overall, the results of the current study underscore the importance of PLA(2) signaling in mercury-induced endothelial dysfunctions. PMID- 20020882 TI - Rapid in vitro screening of drug-metal ion interactions. AB - ABSTRACT The toxic side effects of synthetic drugs may, in part, be arising due to their interactions with essential metal ions, especially when the metal ions are administered along with the drug as mineral supplements. In this paper we report the feasibility of establishing such drug-metal ion interactions through in vitro spectrophotometric studies, which are rapid and can be used for routine screening prior to clinical studies. The interaction of the drugs levothyroxine and ranitidine with eight metal ions, copper(II), chromium(III), molybdenum(VI), magnesium(II), calcium(II), iron(II), manganese(II), and zinc(II), commonly used in mineral supplements, was verified through in vitro UV-visible spectrophotometric studies. The experiments were carried out at the physiological pH values 1.5, 7.4, and 8.0 and the concentrations of the drugs and mineral supplements used were comparable to those in their usual doses. These studies indicated interaction between ranitidine and calcium(II), magnesium(II), and iron(II) ions and between levothyroxine and copper(II) and iron(II) ions. A comparison of the results with those reported from clinical studies demonstrated the efficacy of this method. PMID- 20020883 TI - Ki-67 protein expression in smokeless tobacco (maras powder)-induced oral mucosal lesions. AB - ABSTRACT Objective: Many different forms of smokeless tobacco are used in the world. An interesting kind of smokeless tobacco (Maras powder) is used commonly in lieu of cigarettes in Kahramanmaras and its environs, in Turkey. The object of the present study was to investigate the effects of the Maras powder (MP) on oral mucosa and to examine the expression of Ki-67 in oral mucosa samples taken from MP users. STUDY DESIGN: Expression patterns for markers of cell proliferation (Ki 67) in oral mucosa of 84 MP users and 30 controls were analyzed by immunohistochemistry. RESULTS: Histologically, acanthosis, hyperorthokeratosis, and parakeratosis were found in all 84 formalin-fixed, paraffin-embedded oral mucosal samples of MP users. The spinous cell layer showed large pale staining epithelial cells with pyknotic nuclei. The subepithelial infiltrate was mild. The epithelium was hyperplastic in 17 cases and dysplastic in 67 cases. The dysplasia was mild in 25 cases, moderate in 35 cases, and severe in seven cases. Immunohistochemical findings revealed that with progression of lesions toward malignancy, there was a significant predilection for a basal-suprabasal staining pattern for Ki-67 antigen as compared with the strictly basal staining pattern seen in normal oral epithelium. Expression of Ki-67 antigen was significantly higher in MP users than in the controls (P < 0.001). CONCLUSIONS: Our study has shown that MP may induce epithelial cell proliferation, and that there was a significant positive association of Ki-67 expression and grade of dysplasia in MP users. MP, in contrast to other smokeless tobacco variants, seems to cause mucosal changes, which may transform into oral cancer. PMID- 20020884 TI - The Study of Analgesic Effects and Acute Toxicity of Tripleurospermum disciforme in Rats by Formalin Test. AB - ABSTRACT Previous studies have demonstrated the anti-inflammatory effect of Tripleurospermum disciforme extract in rats with a hind paw edema test. The present study was done to further characterize this anti-inflammatory effect using a formalin test in the intact rats, specifically studying the analgesic effect as well as its toxicity effect by determination of LD50 using graded oral doses of T. disciforme. The effects of graded doses of T. disciforme were investigated following intraperitoneal (IP) administration. Forty-eight rats were used in six groups of studies. The first group (control) received formalin 2.5%, the second group formalin 2.5% + saline, and the third group formalin 2.5% and morphine; three other groups received formalin 2.5% and 15 mg/kg, 30 mg/kg, and 60 mg/kg. After 5, 15, 30, 45, 60 min the pain responses were measured T. disciforme 60 mg/kg had a significant analgesic effect and this effect was equal to morphine 2 mg/kg in the chronic phase of the formalin test. To assess the utility of T. disciforme, the toxic effect of extract was studied by determination of LD50 using graded oral doses of 50, 500, and 5000 mg/kg of T. disciforme to fasted rats and LD50 was measured after 24, 48, and 72 hours. In conclusion, these studies show that T. disciforme is significantly nontoxic at doses required as analgesic drug and higher doses of T. disciforme could produce significant analgesic effects and confirm its previous significant anti inflammatory effect. PMID- 20020885 TI - Importance of drug transporters in pharmacokinetics and drug safety. AB - ABSTRACT The exact role of transporters in the absorption, distribution, and excretion (ADE) of drugs is still being elucidated; however, current evidence suggests that their role in these processes may be significant. In this review, the role of transporters in the ADE, pharmacology, toxicology, and clinical drug drug interactions (DDIs) of drugs will be discussed. The current biological tools to assess this role and practical perspectives for the utilization of knowledge about drug transporters to drug discovery and development will also be discussed. Currently, regulatory agencies are focusing on these potential transporter mediated DDIs. Therefore, guidance and caution on the investigation of these DDIs will be given. PMID- 20020886 TI - Increased Antioxidant Potential and Decreased Free Radical Production in Response to Mild Injection of Crude Venom, Cerastes cerastes gasperetti. AB - ABSTRACT The purpose of this study was to examine the effects of Cerastes cerastes gasperetti crude venom (CCV) on free radical (FR) production and biological antioxidant potential (BAP). Thirty male rats, matched with age and weight, were divided randomly into three groups: control (C), negative control (NC), and experimental (E). Animals in the control group served as control for needlestick and venom injection. Animals in the negative control group, which served as control for venom-induced effects, were injected intraperitoneally (IP) with saline for 8 days, whereas animals of the experimental group were injected IP with crude CCV, at a dose of 1/4 LD(50), for the same period. All animals of the three groups remained under normal laboratory and dietary conditions. Blood samples were collected by heart puncture technique and were analyzed immediately for FRs and BAP. ANOVA analysis indicated a significant difference (p < 0.05) among group means. Tukey-Kramer multiple comparisons showed that FRs increased significantly (p < 0.05) in the NC group, implying that needlestick during injection of saline for 8 days caused internal organ injury sufficient enough to induce reactive oxygen species (ROS) generation. Surprisingly, there were no significant (p > 0.05) changes in FRs in the E group as compared with the C group, which indicates that CCV counteracted the effects induced by ROS. BAP increased significantly (p < 0.05) in the E group as compared with the NC group, reflecting the activation of some antioxidant enzymes. Linear regression of FRs on BAP was significant (p < 0.05) in both the C and E groups. Based on the results of this study, it can be concluded that the CCV contains biochemical factors that activate antioxidant enzymes, as evident by elevation in the BAP that depresses ROS formation. PMID- 20020887 TI - Investigations of oxidative stress, antioxidant response, and protein binding in chlorpyrifos exposed rat neuronal PC12 cells. AB - ABSTRACT Chlorpyrifos (CPF) is a widely used organophosphate insecticide. In addition to its known properties of cholinesterase inhibition, the production of reactive oxygen species (ROS) has been suggested as a possible toxic mechanism. To investigate CPF-generated ROS, rat neuronal PC12 cells were exposed to CPF concentrations of 0 to 5000 mug/mL in Krebs buffered media (KRH), KRH + 4% bovine serum albumin (BSA), and KRH + 25 muM of the antioxidant Trolox for 0 to 5 h. Paraquat served as a positive control for ROS. The fluorescent probe 2,7 dichlorodihydro-fluorescein and the MTS assay were used to measure ROS and cytotoxicity, respectively. Examinations into CPF-albumin binding were also conducted. CPF was not strongly cytotoxic to PC12 cells, causing only mild cytotoxicity at 5000 mug/ml. In KRH media, CPF-generated ROS was observed at 4 and 5 h at 500 and 1000 mug/mL, and at 1 to 5 h at 5000 mug/mL CPF. In KRH + 4% BSA, ROS was seen only at 5 h in 5000 mug/mL CPF. Trolox significantly reduced CPF- and paraquat-induced ROS. Calculated CPF-albumin binding at 1, 10, and 100 mug/mL CPF in 4% BSA was 96%, 75%, and 15%. These data show CPF at >/=500 mug/mL induced ROS in PC12 cells, but the addition of the antioxidant Trolox and 4% BSA dramatically reduced ROS levels. PMID- 20020888 TI - Effects of tributyltin on protein tyrosine kinases and phospholipase C gamma in human natural killer cells. AB - ABSTRACT Tributyltin (TBT) has been used in wood preservation, marine antifouling paints, disinfection of circulating industrial cooling waters, and slime control in paper mills. Detectable levels have been found in human blood. Exposure to TBT decreases the tumor cell lysing (lytic) function of human natural killer (NK) lymphocytes. In this study we assessed the effects of concentrations of TBT that have been shown to decrease NK lytic function on protein tyrosine kinases (PTKs) (Syk, Zap-70, Src, and Pyk) and phospholipase C gamma (PLC-gamma) in NK cells. Exposure to 500 nM TBT caused no change in phosphorylation of any of the PTKs. A 60-min exposure of NK cells to 500 nM TBT did not significantly affect the phosphorylation state of PLC-gamma at any of the lengths of exposure. However, total levels of PLC-gamma were increased by almost 50% after this exposure. Exposure of NK cells to 300 nM TBT for 5 to 60 min caused no significant changes in the phosphorylation state PTKs or PLC-gamma. Exposure of NK cells to 200 nM TBT for 24 h caused no significant changes in the PTK phosphorylation state or total levels. Cells that were exposed to 300 nM TBT for 1 h followed by 24 h or 48 h in TBT-free media showed a significant increase in the phosphorylated forms of Syk and Zap-70 after 24 h in TBT-free media but not after 48 h. These data indicate that in vitro exposure to TBT caused no changes in PTK or PLC-gamma phosphorylation under most conditions. PMID- 20020889 TI - Auricular chondropathy in rats at a contract research toxicology laboratory. AB - ABSTRACT The veterinary staff at WIL Research Laboratories, LLC, was asked to assess and characterize a condition of swollen and reddened ears that had been observed in some of the rats in our facility. When all of the Crl:CD(SD) rats in the facility were examined in August of 2004, August of 2005, and February of 2007, the incidences were 0.25%, 0.18%, and 0.19%, respectively. The overall incidence was 0.21%, with a total of 51 of 24677 animals affected. Only bilaterally affected animals were considered positive. Incidence was strongly correlated with the presence of an ear tag. All but one of the affected animals had an ear tag present. Severe auricular chondropathy was diagnosed on microscopic exam. PMID- 20020890 TI - Serum Profiling of Rat Dermal Exposure to JP-8 Fuel Reveals an Acute-Phase Response. AB - ABSTRACT Dermal exposure to JP-8 petroleum jet fuel leads to toxicological responses in humans and rodents. Serum profiling is a molecular analysis of changes in the levels of serum proteins and other molecules in response to changes in physiology. This present study utilizes serum profiling approaches to examine biomolecular changes in the sera of rats exposed to dermal applications of JP-8 (jet propulsion fuel-8). Using gel electrophoresis and electrospray ionization (ESI) mass spectrometry (MS), levels of serum proteins as well as low mass constituents were found to change after dermal exposures to JP-8. The serum protein levels altered included the acute-phase response proteins haptoglobin, ceruloplasmin, alpha(1)-inhibitor III, and apolipoprotein A-IV. Haptoglobin levels increased after a 1-day JP-8 dermal exposure and continued to increase through 7 days of exposure. Ceruloplasmin levels increased after 5 days of exposure. Serum alpha(1)-inhibitor III was reduced after a 1-day exposure and the depletion continued after 7 days of exposure. Apolipoprotein A-IV increased after a 1-day exposure and then returned to basal levels after 3- and 5-day exposures of JP-8. Levels of the acute-phase protein alpha(2)-macroglobulin were found to not vary over these time course studies. Using ESI-MS analysis directly on the sera from rats exposed to dermal JP-8, low-mass sera constituents were found to correlate with control (acetone) or JP-8 exposure. PMID- 20020891 TI - The effect of etoricoxib, a cyclooxygenase-2-specific inhibitor, on the 1,2 dimethylhydrazine-administered rat intestinal membrane structure and function. AB - ABSTRACT To gain insight into the chemopreventive effects of etoricoxib, which is a selective inhibitor of cycloxygenase-2, a study was carried out in the procarcinogen 1,2-dimethylhydrazine-treated rat intestine. The male Sprague Dawley rats were divided into three different groups. Group 1 served as control (vehicle treated). All animals in Group 2 were given a weekly subcutaneous injection of 1,2-dimethylhydrazine (DMH; 30 mg/kg body weight) for 6 weeks. Group 3 animals were given an additional oral dose of etoricoxib (6 mg/kg body weight) along with weekly DMH injections for 6 weeks. At the end of 6 weeks of treatments, the results indicated significant alterations in the biochemical parameters, membrane lipid composition, and membrane fluorescence studies of the intestine in the presence of DMH, which were recovered nearly to the control level and, therefore, may suggest the chemopreventive efficacy of etoricoxib against the experimental intestinal cancer in rats. PMID- 20020892 TI - Modulation of spontaneous and lipopolysaccharide-induced nitric oxide production and apoptosis by d-galactosamine in rat hepatocyte culture: the significance of combinations of different methods. AB - ABSTRACT Apoptotic markers and signals produced by xenobiotics as hepatotoxic D galactosamine (D-GalN) and lipopolysaccharide (LPS) are extensively investigated in vivo. The contribution of various cells and factors as nitric oxide (NO) in mediating hepatocyte apoptosis in a rat model of systemic endotoxemia was reported. Therefore, the aim of the present work was to study the in vitro effect of D-GalN on nonstimulated or LPS-treated rat hepatocytes in culture and the potential involvement of NO in this process. Our results showed that the spontaneous and LPS-induced NO production was completely blocked by D-GalN during 0 to 24 hours. However, D-GalN slightly enhanced NO production during 24 to 48 hours. D-GalN was more potent to induce hepatocyte apoptosis and necrosis during 24 to 48 than 0 to 24 hours as evidenced morphologically (Annexin V/propidium iodide staining) and biochemically (caspase-3-like activity, alanine aminotransferase leakage, MTT test). Interestingly, D-GalN treatment suppressed mitochondrial cytochrome C release throughout the study. LPS addition to D-GalN considerably aggravated apoptotic/necrotic markers only during 0 to 24 hours. Surprisingly, a share of apoptotic cells was distinctly lower after LPS + GalN treatment than after LPS alone during 0 to 24 hours, while 24- to 48-hour incubation produced massive apoptotic/necrotic hepatocytes. It may be concluded that there is a significant modulation of NO production by D-GalN. Because the role of NO is only partly decisive in the apoptotic/necrotic events, and considering the fraction of the cells completing apoptosis while others that turn toward necrosis (aponecrosis), caution should be exercised in apoptosis data interpretation and combinations of different test methods should be applied. PMID- 20020894 TI - Metabolic markers of hypoxia: systems biology application in biomedicine. AB - ABSTRACT The overall goal of this review is to provide insight into methodologies for 'omic investigations and hypoxic biomarkers that have been identified using 'omic techniques. First, a detailed description of current metabolomic, proteomic, and genomic technologies is provided, followed by a basic introduction to biostatistics and how to interpret 'omic data. Metabolomic biomarkers of diseases in which hypoxia plays a role are then reviewed by those that involved chronic (pulmonary disease, cardiovascular disease, cancer) and acute (stroke, myocardial infarction, ischemia) hypoxia. Data are presented with consideration for the source of hypoxia, the severity of hypoxia, the length of hypoxia, and the cell or organ affected, all of which can have significant effects on biomarker profiles. Drugs that promote and antagonize hypoxia are discussed and important points to consider during tissue collection in hypoxia 'omic studies are then reviewed. PMID- 20020893 TI - Studying the effects of lead on DNA fragmentation and proapoptotic bax and antiapoptotic bcl-2 protein expression in PC12 cells. AB - ABSTRACT The nervous system is one of the most important targets of lead poisoning. Despite decades of study, the exact mechanism of lead toxicity has not been fully elucidated. One of the suggested mechanisms of lead toxicity is induction of apoptosis, which has not been shown yet in some neuronal cells such as pheochromocytoma cells (PC12). Therefore, the present study sought to examine the effect of lead poisoning on apoptosis in PC12 cells as a suitable model of neuronal cell study. The present results showed that lead could induce toxicity in PC12 cells after 24 hours with as little as 1 muM in a concentration-dependent manner. In Western blot analysis, the ratio of Bax/Bcl-2 protein expression in cells incubated with 3, 30, and 90 muM lead acetate significantly increased compared to controls. Additionally, a DNA laddering pattern in lead-treated cells was shown, which could indicate nuclear fragmentation. It might be concluded that lead could cause PC12 cell death, in which apoptosis or programmed cell death plays an important role. PMID- 20020895 TI - The role of metabolic biomarkers in drug toxicity studies. AB - ABSTRACT Metabolic profiling is a technique that can potentially provide more sensitive and specific biomarkers of toxicity than the current clinical measures benefiting preclinical and clinical drug studies. Both nuclear magnetic resonance (NMR) and mass spectrometry (MS) platforms have been used for metabolic profiling studies of drug toxicity. Not only can both techniques provide novel biomarker(s) of toxicity but the combination of both techniques gives a broader range of metabolites evaluated. Changes in metabolic patterns can provide insight into mechanism(s) of toxicity and help to eliminate a potentially toxic new chemical entity earlier in the developmental process. Metabolic profiling offers numerous advantages in toxicological research and screening as sample collection and preparation are relatively simple. Further, sample throughput, reproducibility, and accuracy are high. The area of drug toxicity of therapeutic compounds has already been impacted by metabolic profiling studies and will continue to be impacted as new, more specific biomarker(s) are found. In order for a biomarker or pattern of biomarkers to be accepted, it must be shown that they originate from the target tissue of interest. Metabolic profiling studies are amenable to any biofluid or tissue sample making it possible to link the changes noted in urine for instance as originating from renal injury. Additionally, the ease of sample collection makes it possible to follow a single animal or subject over time in order to determine whether and when the toxicity resolves itself. This review focuses on the advantages of metabolic profiling for drug toxicity studies. PMID- 20020896 TI - VX Penetration Following Percutaneous Poisoning: A Dermal Microdialysis Study in the Guinea Pig. AB - ABSTRACT VX, a potent organophosphorus compound, acts primarily by irreversibly inhibiting acetylcholinesterase resulting in an accumulation of acetylcholine, which produces the characteristic signs of nerve agent poisoning. VX is a low volatility agent, and therefore the most likely route of absorption into the body is via the skin. This study demonstrates for the first time that it is possible to follow the time course of percutaneous VX penetration using the technique of dermal microdialysis and that VX is absorbed through the skin of the anesthetized guinea pig in a concentration-dependent manner. A linear microdialysis probe (5 kDa cut-off) was implanted in the dermis of the back of the guinea pig and perfused (5 muL/min) with physiological Ringer's solution. VX (296 or 592 mug/kg) was applied (33 muL/kg) over the site of the microdialysis probe and dialysate samples collected for up to 6 h. The VX dialysate concentration was measured by liquid chromatography-tandem mass spectrometry (LC-MS-MS). Quantitation was performed over the range 0.1 to 100 ng/mL and the calibration was linear. VX was detected within 15 min, reaching a peak at 30 min following both VX doses. After this time the VX concentration decreased. There was a clear dose-dependent recovery of VX in the dialysate and the total amount recovered was statistically significant between the two doses. This study has clearly shown that microdialysis can be used to follow the time course of the percutaneous absorption of VX in the anesthetized guinea pig and will be used in future studies to develop improved medical countermeasures. Crown Copyright (c) 2007 Dstl. PMID- 20020897 TI - Toxicity Studies of Polynuclear Aromatic Hydrocarbons (PAHs) on European Amphipods. AB - ABSTRACT The effect of phenanthrene, fluoranthene, and pyrene in dimethyl sulfoxide on the amphipods Gammarus aequicauda, Gammarus locusta, and Corophium multisetosum was tested in a static exposure in sea water. The 48-h lethal concentration (LC(50)) of phenanthrene was 173.85 mug/L for G. aequicauda, 147.64 mug/L for G. locusta, and 215.20 mug/L for C. multisetosum. The 48-h LC(50) of fluoranthene was 49.99 mug/L for G. aequicauda, 42.71 mug/L for G. locusta, and 2.85 mug/L for C. multisetosum. The 48-h LC(50) of pyrene was 73.49 mug/L for G. aequicauda, 60.78 mug/L for G. locusta, and 25.29 mug/L for C. multisetosum. Together with their wide distribution along European coasts, the evidence of toxicity on the tested PAH compounds in these amphipods make these species appropriate candidates for evaluating oil-contaminated sediments in Europe. PMID- 20020898 TI - In Vitro Comparison of Two Most Promising H-Oximes (HI-6 and HLo-7) and Currently Commercially Available Reactivators Pralidoxime and Obidoxime in Reactivation of Cyclosarin-Inhibited Human Cholinesterases. AB - ABSTRACT This study describes the evaluation of the in vitro ability of two acetylcholinesterase (EC 3.1.1.7) reactivators, HI-6 and HLo-7, very promising at present, to reactivate human brain cholinesterases inhibited by the nerve agent cyclosarin. The results obtained (percentage of reactivation and appropriate constants characterizing the whole reactivation process) were compared with two currently available reactivators on the market: pralidoxime and obidoxime. It is clear that both promising oximes surpassed the potency of standard reactivators, especially at human relevant concentrations (10(-4) M and lower). Because of the prohibition of such experiments on humans, data obtained in this study could be used as input data for prediction of in vivo action of these drugs in future. PMID- 20020899 TI - Inflammatory and neurochemical changes associated with 3-nitropropionic Acid neurotoxicity. AB - ABSTRACT Administration of 3-nitropropionic acid (3-NP), a hemotoxin of fungal origin, is associated with a rare neurodegenerative disease, Huntington disease. The present study was carried out to investigate the inflammatory and neurochemical changes associated with the subchronic administration of 3 nitropropionic acid in rats. The administration of 3-nitropropionic acid (20 mg/kg SC and 15 mg/kg IP for 7 days) resulted in a significant increase in inflammatory mediators such as tissue necrosis factor (TNF)-alpha, total nitric oxide, and nitrite levels in brain homogenate. It also resulted in a significant decrease in catecholamine neurotransmitters such as dopamine and serotonin (5-HT) and an increase in turnover of these neurotransmitters as compared to the control group, estimated by HPLC with ECD detector. Inflammatory and neurochemical changes were in accordance to the behavioral and biochemical changes induced by 3 NP. The present study suggests the involvement of inflammatory changes as an important causal factor for neurodegeneration associated with 3-NP. PMID- 20020900 TI - Cytotoxicity and oxidative damage in kidney cells exposed to the mycotoxins ochratoxin a and citrinin: individual and combined effects. AB - ABSTRACT Ochratoxin A (OTA) and citrinin (CTN) are two mycotoxins, quite common contaminants, that can occur jointly in a wide range of food commodities. Both mycotoxins have several toxic effects but both share a significant nephrotoxic potential since OTA and CTN were reported to be responsible for naturally occurring human and animal kidney diseases. Considering the concomitant production of OTA and CTN, it is very likely that humans and animals are always exposed to the mixture rather than to individual compounds. Therefore, the aim of the present study was to investigate, using kidney cell culture (Vero cells), whether cytotoxicity and essentially oxidative cell damage (a key determinant of renal diseases) are enhanced by combination of both mycotoxins as compared to their effect separately. To this end, we have assessed their effects individually or combined on cell proliferation using three different cell viability assays (MTT, Trypan Blue, and Neutral Red). In addition, the role of oxidative stress was investigated by measuring the malondialdehyde (MDA) level and the expression of the heat shock protein Hsp 70. Our results clearly showed that cultured renal cells respond to OTA and CTN exposure by a moderate and weak inhibition of cell proliferation and induction of oxidative stress, respectively. However, when combined, they exert a significant increase in inhibition of cell viability as well as the induction of MDA level and Hsp 70 expression. OTA and CTN combination effects are clearly of synergistic nature. The enhanced induction of oxidative stress observed with OTA and CTN simultaneously could be relevant to explain the molecular basis of the renal diseases induced by these mycotoxins. PMID- 20020901 TI - Validation of an Analytical Methodology for Determination of Oxytetracycline Residue in Milk by HPLC with UV Detection. AB - ABSTRACT Oxytetracycline (OTC) is used for the prophylaxis and treatment of a great number of diseases since this antibiotic possesses broad-spectrum activity against many pathogenic organisms. The use of OTC has become a serious problem because of the possible existence of its residues in milk, which can be directly toxic or cause allergic reactions in some hypersensitive individuals. Even low level doses of antibiotic in milk consumed for long periods can lead to problems regarding the spread of drug-resistant microorganisms. The purpose of the present study was to investigate residual OTC in consuming milk in Tehran using high performance liquid chromatography (HPLC) with UV detector. OTC residues in extracts obtained from a preliminary cleanup procedure and recoveries from spiked OTC in desire concentrations were between 80% and 97% with appropriate coefficients of variation. The limit of detection (LOD) and limit of determination (LOQ) were 50 and 68.5 ng/mL, respectively. This result shows that this method would be useful for routine monitoring of oxytetracycline residues in bovine dairy milk. PMID- 20020902 TI - Preliminary studies of sulphur mustard-induced lung injury in the terminally anesthetized pig: exposure system and methodology. AB - ABSTRACT Although normally regarded as a vesicant, inhalation of sulphur mustard (HD) vapor can cause life-threatening lung injury for which there is no specific treatment. Novel therapies for HD-induced lung injury are best investigated in an in vivo model that allows monitoring of a range of physiological variables. HD vapor was generated using two customized thermostatically controlled glass flasks in parallel. The vapor was passed into a carrier flow of air (81 L. min(-1)) and down a length of glass exposure tube (1.75 m). A pig was connected to the midpoint of the exposure tube via a polytetrafluoroethylene-lined endotracheal tube, Fleisch pneumotachograph, and sample port. HD vapor concentrations (40 122.8 mg. m(-3)) up-and downstream of the point of exposure were obtained by sampling onto Porapak absorption tubes with subsequent analysis by gas chromatography-flame photometric detection. Real-time estimates of vapor concentration were determined using a photo-ionization detector. Lung function indices (respiratory volumes, lung compliance, and airway resistance) were measured online throughout. Trial runs with methylsalicylate (MS) and animal exposures with HD demonstrated that the exposure system rapidly reached the desired concentration within 1 min and maintained stable output throughout exposure, and that the MS/HD concentration decayed rapidly to zero when switched off. A system is described that allows reproducible exposure of HD vapor to the lung of anesthetized white pigs. The system has proved to be robust and reliable and will be a valuable tool in assessing potential future therapies against HD induced lung injury in the pig. Crown Copyright (c) 2007 Dstl. PMID- 20020903 TI - Application of benchmark dose for occupational epidemiology in lead exposure. AB - ABSTRACT The benchmark dose (BMD) method has been proposed as an alternative to the no-observed-adverse-effect level (NOAEL) approach for assessing noncancer risks associated with hazardous compounds. The benchmark dose method is a more powerful statistical tool than the traditional NOAEL approach and represents a step in the right direction for a more accurate risk assessment. The benchmark dose method involves fitting a mathematical model to all the dose-response data within a study, and thus more biological information is incorporated in the resulting estimates of guidance values. The BMD and the lower confidence limit on BMD (BMDL) of blood lead to cause renal dysfunction were determined in the population exposure to lead. The blood lead level was used as an exposure biomarker, while total protein (TP), beta(2)-microglobulin (beta(2)-MG), and N Acetyl-beta-D-glucosaminidase (NAG) in the urine were considered as effect biomarkers. The dichotomized data were used as effect endpoints. The BMD and BMDL of blood lead were determined at the 10% benchmark response for the effect biomarkers by using BMDS Version 1.3.1. The results showed that BMD and BMDL of blood lead for NAG, TP, and beta(2)-MG ranged from 323.6 to 754.3 mug/L and 274.2 to 541.5 mug/L, respectively. The BMDL for blood lead was ranked from high to low as TP, beta(2)-MG, and NAG. Urinary NAG activity could be served as a sensitive indicator to detect early renal dysfunction. PMID- 20020904 TI - Microarray analysis of the toxicogenomics and the genotoxic potential of a cationic lipid-based gene delivery nanosystem in human alveolar epithelial a549 cells. AB - ABSTRACT Viral and nonviral vectors have been widely used in gene therapy as delivery reagents for nucleic acids. Toxicity with viral vectors has increasingly led to the search for suitable nonviral vectors, such as cationic lipids/polymers, as potentially safer alternatives. However, little is known about the genomic toxicity of these delivery systems in target cells/tissues. In the current investigation, we report on the toxicogenomics and genotoxicity of cationic lipid Oligofectamine (OF) nanosystems in human alveolar epithelial A549 cells. To investigate the nature and the ontology of the gene expression changes in A549 cells upon treatment with OF nanoliposomes, microarray gene expression profiling methodology was utilized. For microarray analysis, cyanine (Cy3/Cy5) labeled cDNA samples from treated and untreated cells were hybridized on target arrays housing 200 genes. Both OF and OF-DNA lipoplex induced significant gene expression changes belonging to the different genomic ontologies such as cell defense and apoptosis pathways. Flow cytometry analyses revealed induction of apoptosis in A549 cells treated with these nanosystems that is likely due to interactions and/or deterioration of the cell membranes. However, no DNA damage was detected by the Comet assay. These data suggest that cationic nanoliposomes in the absence of direct DNA damage elicit multiple gene expression changes in A549 cells that may compromise the main goals of gene medicine where only therapy defined gene changes are required. PMID- 20020905 TI - Role of nitric oxide synthesis inhibitors in iron-induced nigral neurotoxicity: a mechanistic exploration. AB - ABSTRACT In the central nervous system, nitric oxide (NO) has been suggested to be a cell-to-cell signaling molecule that regulates guanylyl cyclase, aconitase, and iron regulatory protein. NO is also one of the substances that is involved in neuronal death. On the other hand, iron overload and enhanced hydroxyl radical formation have been implicated as the causative factors of some neurodegenerative disorders. The present study was performed to clarify whether nitric oxide is involved in iron-induced neuron death. Neurotoxicity was produced by microinjection of iron chloride (200 mM, 2.5 muL) into the left cerebral ventricle. After the intracerebroventricular (ICV) injection, all animals were kept alive for 10 days. During this period, animals in the iron + L-NAME (N-nitro L-arginine methyl ester) and iron + aminoguanidine groups received intraperitoneal (IP) L-NAME (30 mg/kg) and aminoguanidine (100 mg/kg) injections once a day, respectively. Rats belonging to the control group also received intraperitoneally the same amount of saline. After 10 days, the rats were perfused intracardially under deep urethane anesthesia. Removed brains were processed using the standard histological techniques. The total numbers of neurons in substantia nigra of all rats were estimated with stereological techniques. It was found that L-NAME significantly decreased nigral cell loss from 43.2% to 14.0%, while aminoguanidine did not affect cell loss. Results of the present study suggest that NOS inhibition by L-NAME seems to have neuroprotective effects on iron-induced nigral neurotoxicity. PMID- 20020907 TI - Foreword. PMID- 20020906 TI - Computational toxicology. PMID- 20020908 TI - Toxicity data informatics: supporting a new paradigm for toxicity prediction. AB - ABSTRACT Chemical toxicity data at all levels of description, from treatment level dose response data to a high-level summarized toxicity "endpoint," effectively circumscribe, enable, and limit predictive toxicology approaches and capabilities. Several new and evolving public data initiatives focused on the world of chemical toxicity information-as represented here by ToxML (Toxicology XML standard), DSSTox (Distributed Structure-Searchable Toxicity Database Network), and ACToR (Aggregated Computational Toxicology Resource)-are contributing to the creation of a more unified, mineable, and modelable landscape of public toxicity data. These projects address different layers in the spectrum of toxicological data representation and detail and, additionally, span diverse domains of toxicology and chemistry in relation to industry and environmental regulatory concerns. For each of the three projects, data standards are the key to enabling "read-across" in relation to toxicity data and chemical-indexed information. In turn, "read-across" capability enables flexible data mining, as well as meaningful aggregation of lower levels of toxicity information to summarized, modelable endpoints spanning sufficient areas of chemical space for building predictive models. By means of shared data standards and transparent and flexible rules for data aggregation, these and related public data initiatives are effectively spanning the divides among experimental toxicologists, computational modelers, and the world of chemically indexed, publicly available toxicity information. PMID- 20020909 TI - Computational toxicology methods in public health practice. AB - ABSTRACT Hazard identification and health risk assessment traditionally rely on results of experimental testing in laboratory animals. It is a lengthy and expensive process, which at the end still involves large uncertainty because the sensitivity of animals is unequal to the sensitivity of humans. The Agency for Toxic Substances and Disease Registry (ATSDR) Computational Toxicology and Method Development Laboratory develops and applies advanced computational models that augment the traditional toxicological approach with multilevel cross extrapolation techniques. On the one hand, these techniques help to reduce the uncertainty associated with experimental testing, and on the other, they encompass yet untested chemicals, which otherwise would be left out of public health assessment. Computational models also improve understanding of the mode of action of toxic agents, and fundamental mechanisms by which they may cause injury to the people. The improved knowledge is incorporated in scientific health guidance documents of the Agency, including the Toxicological Profiles, which are used as the basis for scientifically defensible public health assessments. PMID- 20020910 TI - Predictivity and reliability of QSAR models: the case of mutagens and carcinogens. AB - ABSTRACT This paper presents the results from an evaluation of the noncommercial, (quantitative) structure-activity relationship ([Q]SAR) models for the prediction of mutagenicity and carcinogenicity, carried out in a collaboration between the European Chemicals Bureau Group on Computational Toxicology and the Italian Istituto Superiore di Sanita'. Local QSAR models for congeneric chemical classes and structure alert (SA) models were investigated. The studied models can be interpreted mechanistically, agree with, and/or support the available scientific knowledge, and exhibit good statistics. These models were subjected to external prediction tests with chemicals not considered by the authors of the models. Local QSARs that estimated the potency of congeneric chemicals were 30% to 70% correct, whereas the models that discriminated active and inactive chemicals had considerably higher accuracy (70% to 100%). In addition, the commonly used statistical internal cross-validation procedures were poorly correlated with external validation statistics. The genotoxic-based SA models had an accuracy of about 65% for rodent carcinogens, and about 75% for Salmonella mutagens. However, the SA models did not discriminate well active and inactive chemicals within individual chemical classes, and are more suited for preliminary or large-scale screenings. Overall, the (Q)SAR-based predictions are able to significantly enrich the target of safer chemicals, contribute to the organization and rationalization of data, elucidate mechanisms of action, and complement data from other sources. PMID- 20020911 TI - The Use of (Q)SAR Methods in the Context of REACH. AB - ABSTRACT The new European REACH (Registration, Evaluation, Authorisation of Chemicals) legislation requires new information on chemicals, which is tiered according to production volume. The need for data promotes the use of (quantitative) structure-activity relationships ([Q]SARs) in order to meet the European political goal to protect the lives of animals. Within the preparation for REACH, the EU Commission set up REACH implementation projects (RIPs). They are aimed at giving guidance to industry and regulators on how to interpret the legislation. Within the framework of REACH, (Q)SARs are expected to have specific applications in several fields such as raising concerns for chemicals with low production volume for which no (eco)toxicological data have to be submitted, supporting the grouping of chemicals (family approach), and supporting the classification of chemicals as PBT (persistent/bioaccumulative/toxic) or vPvB (very persistent, very bioaccumulative) substances. Currently, the use of (Q)SARs in European regulatory decision making is limited. However, a new European tool (Toxtree) will be made available along with training courses to promote the use of (Q)SAR approaches under REACH. It is expected that the experimental data, to be submitted by industry and made publicly available under REACH, will provide opportunities to improve existing prediction tools and to develop new approaches for (Q)SAR analysis. In the political perspective, the tools will provide support as a rational basis for the intelligent design of new chemicals with sustainable (eco)toxicological properties. PMID- 20020912 TI - MultiCASE Expert Systems and the REACH Initiative. AB - ABSTRACT This article is a review of the MultiCASE Inc. software and expert systems and their use to assess acute toxicity, mutagenicity, carcinogenicity, and other health effects. It is demonstrated that MultiCASE expert systems satisfy the guidelines of the Organisation for Economic Cooperation and Development (OECD) principles and that the portfolio of available endpoints closely overlaps with the list of tests required by REACH. PMID- 20020913 TI - In silico tools for sharing data and knowledge on toxicity and metabolism: derek for windows, meteor, and vitic. AB - ABSTRACT Lhasa Limited is a not-for-profit organization that exists to promote the sharing of data and knowledge in chemistry and the life sciences. It has developed the software tools Derek for Windows, Meteor, and Vitic to facilitate such sharing. Derek for Windows and Meteor are knowledge-based expert systems that predict the toxicity and metabolism of a chemical, respectively. Vitic is a chemically intelligent toxicity database. An overview of each software system is provided along with examples of the sharing of data and knowledge in the context of their development. These examples include illustrations of (1) the use of data entry and editing tools for the sharing of data and knowledge within organizations; (2) the use of proprietary data to develop nonconfidential knowledge that can be shared between organizations; (3) the use of shared expert knowledge to refine predictions; (4) the sharing of proprietary data between organizations through the formation of data-sharing groups; and (5) the use of proprietary data to validate predictions. Sharing of chemical toxicity and metabolism data and knowledge in this way offers a number of benefits including the possibilities of faster scientific progress and reductions in the use of animals in testing. Maximizing the accessibility of data also becomes increasingly crucial as in silico systems move toward the prediction of more complex phenomena for which limited data are available. PMID- 20020914 TI - Combined Use of MC4PC, MDL-QSAR, BioEpisteme, Leadscope PDM, and Derek for Windows Software to Achieve High-Performance, High-Confidence, Mode of Action Based Predictions of Chemical Carcinogenesis in Rodents. AB - ABSTRACT This report describes a coordinated use of four quantitative structure activity relationship (QSAR) programs and an expert knowledge base system to predict the occurrence and the mode of action of chemical carcinogenesis in rodents. QSAR models were based upon a weight-of-evidence paradigm of carcinogenic activity that was linked to chemical structures (n = 1,572). Identical training data sets were configured for four QSAR programs (MC4PC, MDL QSAR, BioEpisteme, and Leadscope PDM), and QSAR models were constructed for the male rat, female rat, composite rat, male mouse, female mouse, composite mouse, and rodent composite endpoints. Model predictions were adjusted to favor high specificity (>80%). Performance was shown to be affected by the method used to score carcinogenicity study findings and the ratio of the number of active to inactive chemicals in the QSAR training data set. Results demonstrated that the four QSAR programs were complementary, each detecting different profiles of carcinogens. Accepting any positive prediction from two programs showed better overall performance than either of the single programs alone; specificity, sensitivity, and Chi-square values were 72.9%, 65.9%, and 223, respectively, compared to 84.5%, 45.8%, and 151. Accepting only consensus-positive predictions using any two programs had the best overall performance and higher confidence; specificity, sensitivity, and Chi-square values were 85.3%, 57.5%, and 287, respectively. Specific examples are provided to demonstrate that consensus positive predictions of carcinogenicity by two QSAR programs identified both genotoxic and nongenotoxic carcinogens and that they detected 98.7% of the carcinogens linked in this study to Derek for Windows defined modes of action. PMID- 20020915 TI - In Silico Screening of Chemicals for Genetic Toxicity Using MDL-QSAR, Nonparametric Discriminant Analysis, E-State, Connectivity, and Molecular Property Descriptors. AB - ABSTRACT Genetic toxicity testing is a critical parameter in the safety assessment of pharmaceuticals, food constituents, and environmental and industrial chemicals. Quantitative structure-activity relationship (QSAR) software offers a rapid, cost-effective means of prioritizing the genotoxic potential of chemicals. Our goal is to develop and validate a complete battery of complementary QSAR models for genetic toxicity. We previously reported the development of MDL-QSAR models for the prediction of mutations in Salmonella typhimurium and Escherichia coli ( Contrera et al. 2005b ); this report describes the development of eight additional models for mutagenicity, clastogenicity, and DNA damage. The models were created using MDL-QSAR atom-type E-state, simple connectivity and molecular property descriptor categories, and nonparametric discriminant analysis. In 10% leave-group-out internal validation studies, the specificity of the models ranged from 63% for the mouse lymphoma (L5178Y-tk) model to 88% for chromosome aberrations in vivo. Sensitivity ranged from a high of 74% for the mouse lymphoma model to a low of 39% for the unscheduled DNA synthesis model. The receiver operator characteristic (ROC) was >/=2.00, a value indicative of good predictive performance. The predictive performance of MDL-QSAR models was also shown to compare favorably to the results of MultiCase MC4PC ( Matthews et al. 2006b ) genotoxicity models prepared with the same training data sets. MDL-QSAR software models exhibit good specificity, sensitivity, and coverage and they can provide rapid and cost-effective large-scale screening of compounds for genotoxic potential by the chemical and pharmaceutical industry and for regulatory decision support applications. PMID- 20020916 TI - Development of a phospholipidosis database and predictive quantitative structure activity relationship (QSAR) models. AB - ABSTRACT Drug-induced phospholipidosis (PL) is a condition characterized by the accumulation of phospholipids and drug in lysosomes, and is found in a variety of tissue types. PL is frequently manifested in preclinical studies and may delay or prevent the development of pharmaceuticals. This report describes the construction of a database of PL findings in a variety of animal species and its use as a training data set for computational toxicology software. PL data and chemical structures were compiled from the published literature, existing pharmaceutical databases, and Food and Drug Administration (FDA) internal reports yielding a total of 583 compounds suitable for modeling. The database contained 190 (33%) positive drugs and 393 (77%) negative drugs, of which 39 were electron microscopy-confirmed negative compounds and 354 were classified as negatives due to the absence of positive reported data. Of the 190 positive findings, 76 were electron microscopy confirmed and 114 were considered positive based on other evidence. Quantitative structure-activity relationship (QSAR) models were constructed using two commercially available software programs, MC4PC and MDL QSAR, and internal cross-validation (10 x 10%) experiments were performed to assess their predictive performance. Performance parameters for the MC4PC model were specificity 92%, sensitivity 50%, concordance 78%, positive predictivity 76%, and negative predictivity 78%. For MDL-QSAR, predictive performance was similar: specificity 80%, sensitivity 76%, concordance 79%, positive predictivity 65%, and negative predictivity 87%. By combining the output of the two QSAR programs, the overall predictive performance was vastly improved and sensitivity could be optimized to 81% without significant loss of specificity (79%). Many of the structural alerts and significant molecular descriptors obtained from the QSAR software were found to be associated with parts of active molecules known for their cationic amphiphilic drug (CAD) properties supporting the hypothesis that the endpoint of PL is statistically correlated with chemical structure. QSAR models can be useful tools for screening drug candidate molecules for potential PL. PMID- 20020917 TI - In silico toxicological screening of natural products. AB - ABSTRACT This study closely examines six well-known naturally occurring dietary chemicals (estragole, pulegone, aristolochic acid I, lipoic acid, 1-octacosanol, and epicatechin) with known human exposure, chemical metabolism, and mechanism of action (MOA) using in silico screening methods. The goal of this study was to take into consideration the available information on these chemicals in terms of MOA and experimentally determined toxicological data, and compare them to the in silico predictive modeling results produced from a series of computational toxicology software. After these analyses, a consensus modeling prediction was formulated in light of the weight of evidence for each natural product. We believe this approach of examining the experimentally determined mechanistic data for a given chemical and comparing it to in silico generated predictions and data mining is a valid means to evaluating the utility of the computational software, either alone or in combination with each other. We find that consensus predictions appear to be more accurate than the use of only one or two software programs and our in silico results are in very good agreement with the experimental toxicity data for the natural products screened in this study. PMID- 20020918 TI - Combined computational metabolite prediction and automated structure-based analysis of mass spectrometric data. AB - ABSTRACT As high-throughput technologies have developed in the pharmaceutical industry, the demand for identification of possible metabolites using predominantly liquid chromatographic/mass spectrometry-mass spectrometry/mass spectrometry (LC/MS-MS/MS) for a large number of molecules in drug discovery has also increased. In parallel, computational technologies have also been developed to generate predictions for metabolites alongside methods to predict MS spectra and score the quality of the match with experimental spectra. The goal of the current study was to generate metabolite predictions from molecular structure with a software product, MetaDrug. In vitro microsomal incubations were used to ultimately produce MS data that could be used to verify the predictions with Apex, which is a new software tool that can predict the molecular ion spectrum and a fragmentation spectrum, automating the detailed examination of both MS and MS/MS spectra. For the test molecule imipramine used to illustrate the combined in vitro/in silico process proposed, MetaDrug predicts 16 metabolites. Following rat microsomal incubations with imipramine and analysis of the MS(n) data using the Apex software, strong evidence was found for imipramine and five metabolites and weaker evidence for five additional metabolites. This study suggests a new approach to streamline MS data analysis using a combination of predictive computational approaches with software capable of comparing the predicted metabolite output with empirical data when looking at drug metabolites. PMID- 20020919 TI - A hierarchical clustering methodology for the estimation of toxicity. AB - ABSTRACT A quantitative structure-activity relationship (QSAR) methodology based on hierarchical clustering was developed to predict toxicological endpoints. This methodology utilizes Ward's method to divide a training set into a series of structurally similar clusters. The structural similarity is defined in terms of 2 D physicochemical descriptors (such as connectivity and E-state indices). A genetic algorithm-based technique is used to generate statistically valid QSAR models for each cluster (using the pool of descriptors described above). The toxicity for a given query compound is estimated using the weighted average of the predictions from the closest cluster from each step in the hierarchical clustering assuming that the compound is within the domain of applicability of the cluster. The hierarchical clustering methodology was tested using a Tetrahymena pyriformis acute toxicity data set containing 644 chemicals in the training set and with two prediction sets containing 339 and 110 chemicals. The results from the hierarchical clustering methodology were compared to the results from several different QSAR methodologies. PMID- 20020920 TI - Characteristics of genomic signatures derived using univariate methods and mechanistically anchored functional descriptors for predicting drug- and xenobiotic-induced nephrotoxicity. AB - ABSTRACT The ideal toxicity biomarker is composed of the properties of prediction (is detected prior to traditional pathological signs of injury), accuracy (high sensitivity and specificity), and mechanistic relationships to the endpoint measured (biological relevance). Gene expression-based toxicity biomarkers ("signatures") have shown good predictive power and accuracy, but are difficult to interpret biologically. We have compared different statistical methods of feature selection with knowledge-based approaches, using GeneGo's database of canonical pathway maps, to generate gene sets for the classification of renal tubule toxicity. The gene set selection algorithms include four univariate analyses: t-statistics, fold-change, B-statistics, and RankProd, and their combination and overlap for the identification of differentially expressed probes. Enrichment analysis following the results of the four univariate analyses, Hotelling T-square test, and, finally out-of-bag selection, a variant of cross-validation, were used to identify canonical pathway maps-sets of genes coordinately involved in key biological processes-with classification power. Differentially expressed genes identified by the different statistical univariate analyses all generated reasonably performing classifiers of tubule toxicity. Maps identified by enrichment analysis or Hotelling T-square had lower classification power, but highlighted perturbed lipid homeostasis as a common discriminator of nephrotoxic treatments. The out-of-bag method yielded the best functionally integrated classifier. The map "ephrins signaling" performed comparably to a classifier derived using sparse linear programming, a machine learning algorithm, and represents a signaling network specifically involved in renal tubule development and integrity. Such functional descriptors of toxicity promise to better integrate predictive toxicogenomics with mechanistic analysis, facilitating the interpretation and risk assessment of predictive genomic investigations. PMID- 20020921 TI - Understanding genetic toxicity through data mining: the process of building knowledge by integrating multiple genetic toxicity databases. AB - ABSTRACT Genetic toxicity data from various sources were integrated into a rigorously designed database using the ToxML schema. The public database sources include the U.S. Food and Drug Administration (FDA) submission data from approved new drug applications, food contact notifications, generally recognized as safe food ingredients, and chemicals from the NTP and CCRIS databases. The data from public sources were then combined with data from private industry according to ToxML criteria. The resulting "integrated" database, enriched in pharmaceuticals, was used for data mining analysis. Structural features describing the database were used to differentiate the chemical spaces of drugs/candidates, food ingredients, and industrial chemicals. In general, structures for drugs/candidates and food ingredients are associated with lower frequencies of mutagenicity and clastogenicity, whereas industrial chemicals as a group contain a much higher proportion of positives. Structural features were selected to analyze endpoint outcomes of the genetic toxicity studies. Although most of the well-known genotoxic carcinogenic alerts were identified, some discrepancies from the classic Ashby-Tennant alerts were observed. Using these influential features as the independent variables, the results of four types of genotoxicity studies were correlated. High Pearson correlations were found between the results of Salmonella mutagenicity and mouse lymphoma assay testing as well as those from in vitro chromosome aberration studies. This paper demonstrates the usefulness of representing a chemical by its structural features and the use of these features to profile a battery of tests rather than relying on a single toxicity test of a given chemical. This paper presents data mining/profiling methods applied in a weight-of-evidence approach to assess potential for genetic toxicity, and to guide the development of intelligent testing strategies. PMID- 20020922 TI - Appendix for special issue on computational toxicology. PMID- 20020923 TI - Panmasala chewing induces deterioration in oral health and its implications in carcinogenesis. AB - ABSTRACT Panmasala containing tobacco was introduced in the Indian market during the 1970s. Panmasala consists of areca nut (betel nut), catechu, lime, cardamom, spices, and unspecified flavoring agents, etc., with tobacco locally known as gutkha or without tobacco (Plain or sada), and consumed abundantly in India and also other parts of the world, predominantly in South East Asian countries. Available studies demonstrate that the habits of chewing panmasala gutkha or plain by students and adolesescents are on the increase, which may lead to deterioration of oral health and other organ systems. Based on the experimental as well as clinical studies available on panmasala as well as on different components of panmasala, this review suggests that it has the potential in causation of various oral diseases such as Oral Sub Mucosis Fibrosis (OSMF) and leucoplakia which may lead to oral cancer. Studies reviewed on these chewing mixtures also reveal that it is likely to be carcinogenic, as tobacco and areca nut have carcinogenic potential and both have encompassing addictive potential leading to dependence on chewing mixture containing areca nut and tobacco. These mixtures might not only lead to cancer but may also affect other organs of the body, including oral hard tissues in the form of dental attrition and sensitivity. There is a need to consider the potential health hazards associated with the habits of these products, especially oral cancer. More research is needed to find out early changes which could be reversible and also intervention measures through education to desist people in indulging in such habits. PMID- 20020924 TI - Effects of Electromagnetic Radiation Use on Oxidant/Antioxidant Status and DNA Turn-over Enzyme Activities in Erythrocytes and Heart, Kidney, Liver, and Ovary Tissues From Rats: Possible Protective Role of Vitamin C. AB - ABSTRACT In this study, the aim was to investigate possible effects of Electromagnetic Radiation (EMR) use on oxidant and antioxidant status in erythrocytes and kidney, heart, liver, and ovary tissues from rats, and possible protective role of vitamin C. For this aim, 40 Wistar albino female rats were used throughout the study. The treatment group was exposed to EMR in a frequency of 900 MHz, the EMR plus vitamin C group was exposed to the same EMR frequency and given vitamin C (250 mg/kg/day) orally for 4 weeks. There were 10 animals in each group including control and vitamin C groups. At the end of the study period, blood samples were obtained from the animals to get erythrocyte sediments. Then the animals were sacrificed and heart, kidney, liver, and ovary tissues were removed. Malondialdehyde (MDA) levels and superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GSH-Px), xanthine oxidase (XO), and adenosine deaminase (ADA) enzyme activities were measured in the tissues and erythrocytes. It was observed that MDA level, XO, and GSH-Px activities significantly increased in the EMR group as compared with those of the control group in the erythrocytes. In the kidney tissues, it was found that MDA level and CAT activity significantly increased, whereas XO and ADA activities decreased in the cellular phone group as compared with those of the control group. However, in the heart tissues it was observed that MDA level, ADA, and XO activities significantly decreased in the cellular phone group as compared with those of the control group. The results suggest that EMR at the frequency generated by a cell phone causes oxidative stress and peroxidation in the erythrocytes and kidney tissues from rats. In the erythrocytes, vitamin C seems to make partial protection against the oxidant stress. PMID- 20020925 TI - Comparative Measurement of In Vitro Paraquat and Aflatoxin B1 Cytotoxicity Using Three Different Cytotoxicity Assays in Pheochromocytoma Cells (PC-12). AB - ABSTRACT Among the herbicides and mycotoxins, paraquat (PQ) and aflatoxin B1 (AFB1) are highly cytotoxic. In this study the toxicity of PQ and AFB1 in the cultured cell were determined using MTT [3-(4, 5-dimethylthiazol-2-yl)-2, 5 diphenyltetrazolium bromide], JG-B (Janus green B), and NR (neutral red) assay by multiwell scanning spectophotometry. JG-B was used not only for the vital staining of mitochondria but also for viability assay and was compared to MTT and NR assay. Various concentrations of paraquat (0.1 mM to 100 mM) and AFB1 (0.001 nM to 10 nM) on the PC-12 cells were investigated. The 50% lethal concentration of toxins (LC50) were determined for PQ (7.70 +/- 2.50, 3.67 +/- 1.53, 4.85 +/- 2.44) and AFB1 (0.16 +/- 0.01, 0.13 +/- 0.04, 0.14 +/- 0.02) as determined by these methods (JG-B, NR, and MTT, respectively). A significant correlation was found among the JG-B and MTT using PQ (r(2) = 0.99, p < 0.05) and significant correlation was also found among the three methods (r(2) = 0.95, 0.93, and 0.92, p < 0.05) using AFB1. No significant correlation was found between JG-B and MTT with NR (r(2) = 0.34 and 0.35, p < 0.05, respectively) using PQ. These results suggest that both methods (MTT assay and JG-B assay) are reliable and are comparable for determining the cytotoxicity. It is concluded that the JG-B assay may be preferable to MTT assay methods because of its simplicity, low cost, sensitivity, and objectivity; in addition, this method takes little time to be done. PMID- 20020926 TI - Antigenotoxic Effects of Curcumin and Piperine Alone or in Combination Against 7,12-Dimethylbenz(a)anthracene Induced Genotoxicity in Bone Marrow of Golden Syrian Hamsters. AB - ABSTRACT The present study investigates the effect of curcumin and piperine alone or in combination against 7,12-dimethylbenz[a]anthracene (DMBA)-induced genotoxicity in the bone marrow of hamsters. The antigenotoxic effect was evaluated by analyzing the frequency of micronucleated polychromatic erythrocytes (MnPCEs) and chromosomal aberrations. Genotoxicity was induced in experimental hamsters by single intraperitoneal injection of DMBA (30 mg/kg b.w). Oral pretreatment of curcumin (80 mg/kg b.w), piperine (50 mg/kg b.w), and curcumin (80 mg/kg b.w) + piperine (50 mg/kg b.w), respectively, for 5 days, significantly reduced the frequency of MnPCEs and the percentage of chromosomal aberrations in the bone marrow of hamsters. The results suggest that cucumin and piperine in combination have a potent antigenotoxic effect as compared to either agent alone in DMBA-induced genotoxicity in golden Syrian hamsters. PMID- 20020927 TI - Estimation of atrazine in the stratum corneum and its toxic effects in skin following topical application to rabbits. AB - ABSTRACT For risk assessment, a workable non-invasive method for the estimation of atrazine in skin was needed. Moreover, the dermato-toxic effects of different concentrations of atrazine have not been studied so far. Fifteen milligrams of 35% solution of atrazine in ethanol was topically applied to each of six different clipped sites on the back of rabbits (n = 6). Each site was tape stripped with 10 consecutive adhesive tapes at 0.5, 1, 2, 3, 4, and 6 h of topical dosing to remove the stratum corneum (SC). Atrazine in tapes was extracted with methanol and analyzed with HPLC. The amount of atrazine detected in the SC at 6 h (1955.79 +/- 47.22 mug) is equivalent to 13.03% of the total applied dose. In dermal toxicity studies, 500 mul of 35%, 70% atrazine in ethanol and a commercial preparation of atrazine (Balance(R)) was dosed on clipped backs of rabbits (n = 4) for 4 days under occluded vs non-occluded conditions. Ethanol alone served as control. On the 5th day, rabbits were euthanized and skin was scored for erythema and then examined microscopically. Significant differences (p < 0.05) in erythema scores were observed with 70% atrazine and Balance(R) as compared to the control under occluded conditions. Significant differences in epidermal thickness and cell layers were observed with Balance(R) and 74% atrazine as compared to control in both dosing conditions. There were non significant differences in erythema, epidermal thickness, or cell layers in occluded vs non-occluded applications of atrazine in ethanol, indicating that atrazine is equally toxic regardless of its application procedures. Moreover, under non-occluded application, very slight erythema was observed but microscopically significant epidermal hyperplasia was noticed. This indicates that even if there are no significant gross skin manifestations of atrazine this can still produce significant damage to the epidermal barrier and hence can result in increase in penetration of self or other toxic substances. PMID- 20020928 TI - Subacute oral toxicity of endosulfan in male new zealand white rabbits. AB - ABSTRACT The present study was conducted using 6 to 8 month old New Zealand white male rabbits (nine rabbits per treatment group). Daily gavages of 3, 1.5, 0.75, or 0 mg endosulfan/kg BW in corn oil resulted in the death of five (55%), three (33%), zero (0%), and zero (0%) rabbits, respectively, in 30 days. All rabbits were monitored for any observable toxic symptoms throughout the experimental period (30 d) and they also were weighed weekly to monitor body weight gain. All deaths occurred within the first 3 weeks and nervous symptoms were observed only for a few minutes before death. Alterations recorded in hematological parameters within the groups (hemoglobin, packed cell volume, and total erythrocyte count) were due to endosulfan exposure. Serum alkaline phosphatase (ALP) and aspartate aminotransferase (AST) levels were significantly elevated in the 3 mg/kg dose group. Gross post-mortem and histopathological changes in various organs (lung, liver, kidney, and testes) of rabbits treated with endosulfan were observed with typical organochlorine dose-dependent signs of toxicity. Although some animals appeared to adjust to relatively high daily doses of endosulfan for 30 days, biochemical and histological evidence indicated varied liver and kidney damage relative to dosage administered to these animals. The current subacute (30 day) study suggested a NOAEL of 0.75 mg endosulfan/kg in New Zealand white rabbits. PMID- 20020929 TI - The Teratogenic Effects of Flurazepam Intake during Organogenesis of the Rat Fetus. AB - ABSTRACT Flurazepam is a long acting drug with sedative, hypnotic, anxiolytic, relaxant, and anti-epileptic properties. The drug is a benzodiazepine and is commonly used by adults, including pregnant women, intentionally or sometimes unintentionally during their pregnancy. It is increasingly used these days, and therefore, it seems a special problem to the clinician treating anxiety disorders in women. It should be noticed that flurazepam exposure during pregnancy may have teratogenic effects on the fetus. Until now, many studies have been conducted on drug side-effects in poisonings, behavioral disorder, and anxiety reactions, but there is no accurate report about the teratogenic effect of flurazepam. In this study, teratogenicity flurazepam intake during pregnancy and its effects on fetus development was investigated. About 30 virgin rats of known age and weight were used. After being pregnant, they were divided into three groups: Negative and positive control group, case groups exposed for 1 to 6 mg/kg/day. The fetuses were first studied macroscopically regarding anomalies, and then histological and histochemically to inspect the defects of tissue organogenesis. Our results show that there was significant difference in the weight and length of the cases compared to the control group. The statistical results indicate that flurazepam intake during the second half of pregnancy can lead to irreversible anomalies. It seems that benzodiazepine therapy among pregnant woman would be better to avoid during the first trimester and multidrug regimens. PMID- 20020930 TI - In vitro and in silico approaches for analyzing the toxicological effect of triptolide on cx43 in sertoli cells. AB - ABSTRACT Triptolide is a diterpene triepoxide isolated from the traditional Chinese medicinal vine Trypterygium wilfordii hook f. (T. wilfordii). It possesses multiple biological activities, such as antitumor, immunosuppression, and antifertility. Previous studies suggested that triptolide might be a potential candidate for post-testicular male contraceptive agent. Nevertheless, the mechanisms of triptolide-induced reproductive toxicity remain unclear. In the present study, the results of reverse-transcription PCR and Western blotting revealed that triptolide inhibited the expression of Cx43 compared with the different effect of estradiol in cultured SCs from male rats. Further computational study revealed that triptolide can bind to the active site of human estrogenic 17beta-hydroxysteroid dehydrogenase and human estrogen receptorbeta. Therefore, our results indicated that male reproductive toxicity induced by triptolide was associated with the effects on intratesticular estrogen levels and estrogen receptors rather than its cytotoxicity. PMID- 20020931 TI - Rat Plasma Oxidation Status After Nigella Sativa L. Botanical Treatment in CCL(4) Treated Rats. AB - ABSTRACT Nigella sativa Linn. (family Ranunculaceae), commonly known as black cumin, is native to the Mediterranean area and has been used for thousands of years as a health and beauty aid. The present study investigated the protective effects of Nigella sativa (NS) extract (NSE) and oil (NSO) on CCl(4)-induced nitrosative stress and protein oxidation in rat. CCl(4) (0.8 mg/kg) was used as an aid for induction of nitrosative stress. In vitro antioxidant potential was tested in the presence of 2,4-dinitrophenylhyrdazine (DPPH) as an organic nitrogen radical. Doses of 0.2, 0.3, and 1 mg/kg of the NS extract and oil were administered to CCL(4)-treated rats for 10 days. At the end of treatment, blood was taken from rats under anesthesia and plasma was separated. The concentration of nitric oxide (NO), total antioxidant power (TAP), carbonyl molecules (CM) as measure of protein oxidation (PO), tumor necrosis factor-alpha (TNF-alpha), and total thiol molecules (TTM) were measured in plasma. In vitro evaluation of antioxidant effects of NSE and NSO showed that the highest antioxidant activity (80%) was observed with the concentration of 10 and 20 mg/ml, respectively, that were equal to vitamin E (200 mg/ml). Administration of CCL(4) increased plasma PO, NO, TNF-alpha and decreased TAP and TTM. Both NSE and NSO showed significant protection against CCl(4)-induced changes in biochemical parameters, but not dose dependently. Doses of 0.3 and 1 mg/kg were more effective than doses of 0.2 mg/kg for both NSE and NSO, but dose of 1 mg/kg was the most effective one. The results indicate the potential of NS in preventing CCL(4)-induced toxic nitrosative stress. It is concluded that NS has marked antioxidant potentials that may be beneficial in alleviating complications of many illnesses related to oxidative/nitrosative stress in humans, but preclinical safety measures should be completed before clinical trials. PMID- 20020932 TI - Determination of Bisphenol A (BPA) in Plasma of Hemodialysis Patients Using Three Methods: LC/ECD, LC/MS, and ELISA. AB - ABSTRACT Bisphenol A (BPA) values were compared in plasma of hemodialysis patients and in recycling solvents using LC/MS, LC/electrochemical detector (ECD), and enzyme linked immunosorbent assay (ELISA). BPA values in the plasma and the solvent were 0-8.4 ng/ml and 0-0.8 ng/ml for LC/ECD, 0-4.9 ng/ml and 0 0.8 ng/ml for LC/MS, and 0-15.5 ng/ml and 0-3.1 ng/ml for ELISA, respectively. There was no significant difference among BPA values both in the plasma and the solvents using three methods. Single correlation coefficients between LC/ECD and LC/MS, LC/ECD and ELISA, and LC/MS and ELISA were, respectively, 0.373 (p < 0.002), 0.347 (p < 0.002), and 0.945 (p < 0.001) in the plasma (n = 68-109) and 0.916 (p < 0.001), 0.431 (p > 0.05), and 0.332 (p > 0.05) in the solvents (n = 19). An unknown substance present in the plasma of patients but not healthy volunteers influenced the LC/ECD values of plasma repeated freezing and thawing. The results indicate that LC/MS and ELISA are appropriate for BPA analysis in plasma and both LC/MS and LC/ECD in the recycling solvents and handling with plasma before analysis is important to the analysis of BPA in patients' plasma using LC/ECD. PMID- 20020933 TI - Experimental inhalation of chlorine gas produced with a different method; effects of N-acetyl cysteine on acute pulmonary damage. AB - ABSTRACT The two most common gas inhalation injuries encountered in emergency departments are carbon monoxide and chlorine inhalations. In this study, chlorine was produced through a method different to the previous experimental models. Rats were subjected to inhale chlorine, after which the effects of N-acetylcysteine on pulmonary damage were evaluated. A total of 50 rats were equally divided into five groups. Group 1 received nothing. Groups 2 and 3 were taken as 6 h, groups 4 and 5 as 24 h control and N-acetylcysteine groups, respectively. Firstly, 200 ppm chlorine gas was given for 20 min. Then, 40 mg/kg N-acetylcysteine was given intraperitoneally. The same procedure with the same dose was repeated 3 h later. The same procedures were applied to the control group but this time saline was used. Tissue samples of lungs were taken. Glutathione levels of the rats in the N acetylcysteine group sacrificed at 24 h were significantly higher than those of the control group. Histopathological evaluation of the pulmonary tissues of the rats sacrificed at 6 and 24 h revealed mild-to-moderate degrees of tissue damage. The degree of tissue damage at 6 h and 24 h N-acetylcysteine group rats was lower than that in the control group. As a result, tissue damage resulting from experimental chlorine inhalation can be alleviated by N-acetylcysteine. This is mainly the result of the antioxidant effects of the N-acetylcysteine. PMID- 20020934 TI - Alcohol poisoning and venous hyperoxia. AB - ABSTRACT Venous PCO(2) and PO(2) in the presence of normal arterial PCO(2) and PO(2) in patients with alcoholic intoxication have not been previously evaluated. The objective of this study was to compare arterial and venous blood gases in patients with alcoholic intoxication and healthy controls. Sixteen patients with alcoholic intoxication and 20 controls underwent simultaneous blood sampling from a radial artery and an antecubital vein for acid-base analysis. Osmolality and ethanol blood concentration was estimated. Elevated venous pO(2) were found in 56% of patients with alcoholic poisoning compared with 15% of controls. A formula was found describing possible arterio-venous shunt accounting for elevated venous pO(2) and enabling calculation of the relevant venous carbon dioxide content and CO(2) product. The values of the venous pO(2) and arterio-venous shunt were more significant in the alcohol group than in controls (p = 0.002, p = 0.001, respectively). Percentage of patients with a-v shunts was significantly higher in the alcohol group (81%) than in controls (25%) (p = 0.002, OR 2.6, 95% CI 0.13 6.52). The relevant venous CO(2) and CO(2) product had the non-significant trend to be higher in the alcohol group. In conclusion, this study reports ethanol induced venous pO(2) and pCO(2) elevation. This may be associated with the effects of tissue perfusion stealing and high oxygen consumption. On the other hand, possible beneficial consequences may occur: acceleration of alcohol elimination and reduction of alcohol-induced tissue damage. PMID- 20020935 TI - Lead-induced stress response in endoplasmic reticulum of astrocytes in CNS. AB - ABSTRACT Lead is one of the most widespread toxicants in the environment, and its neurotoxicity contributes to a major medical issue. Numerous studies have shown that astrocytes are the main sites of Pb deposition in the central nervous system. A large amount of lead depositing in the astrocyte cells would result in the accumulation of unfolded protein in the endoplasmic reticulum (ER), which up regulates the expression of molecular chaperones and meanwhile inhibits the cell cycle progression and the transcription of certain proteins. The unfolded protein response (UPR) could down-regulate the expression of protein cyclinD1 and cause the stagnation of cell-cycle in primary-cultured astrocytes of rat. However, lead neither has obvious effects on the expression of C/EBP homologous protein (CHOP) nor achieves cell apoptosis in the progress of lead-induced UPR. When the stagnation of cell-cycle happens, glucose regulated protein of 78 kDa (GRP78) and other chaperones come to themselves to transport a body of unfolded-protein, consequently making cells survive. PMID- 20020936 TI - Effect of butyl benzyl phthalate on early postnatal mortality in rats. AB - ABSTRACT The present study was undertaken to examine early postnatal mortality in rat pups following exposure to butyl benzyl phthalate (BBP) during pregnancy. Seventeen pregnant rats were given 750 mg/kg bw/day of BBP by oral gavage on gestation days 13, 14, and 15, and the volume of each dose was adjusted to 5 ml/kg body weight. Four rats were given olive oil only and served as control. Natural birth was allowed to take place. One hundred and eighty-three pups were born to the experimental rats and 46 pups to the control group. Close observation of the newborn pups during the first 3 h of life revealed that all the pups in both the control and experimental groups were born alive. Only six pups from the experimental group (3.2%) died within this time period. These and four control pups were fixed and decalcified. Histological examination of the thoracic cavity of the newborn rats in both groups revealed no differences in the position or size of any of the heart chambers, ductus arteriosus, or great vessels. However, the lungs of the six experimental pups that died showed athelectasia and bronchi dilatation. The results therefore suggest that exposure to BBP of rats during pregnancy does not produce significant postnatal mortality in their offspring. PMID- 20020937 TI - Role of ABC Transporters in the BeWo Trophoblast Cell Line. AB - ABSTRACT The transport of doxorubicin and rhodamine 123, substrates of ABC transporters, was evaluated in the BeWo stabilized trophoblast cell line. Both compounds were taken up by BeWo cells, but their intracellular concentrations were highly dependent on temperature, and significantly reduced at 4 degrees C. The P-glycoprotein inhibitors verapamil and PSC833 did not modify the intracellular concentrations of the two substrates, suggesting therefore that, in these cells, the activity of P-glycoprotein is not important. MK571, which inhibits MRPs, was on the contrary effective in increasing rhodamine 123 intracellular concentrations. The efflux of both fluorescent substrates was extremely slow, and slightly reduced by MK571. Finally, a polarized transport of doxorubicin from basal to apical side was evident, although only during the first 60 min of incubation, and was reduced by P-glycoprotein, MRP, and BCRP inhibitors. No MDR1 expression was revealed at the mRNA and protein levels; on the contrary, MRP1 and BCRP were expressed in these cells. In BeWo cells the activity of ABC transporters, and in particular of P-glycoprotein, seems to be extremely limited. PMID- 20020938 TI - CAS Enhances Chemotherapeutic Drug-Induced p53 Accumulation and Apoptosis: Use of CAS for High-Sensitivity Anticancer Drug Screening. AB - ABSTRACT Cells attacked by cytotoxic toxins may express apoptosis-related proteins such as p53 to kill themselves, so as not to affect surrounding healthy cells. These apoptosis-related proteins are also crucial for inducing apoptosis of tumor cells in cancer chemotherapy. CSE1L/CAS is a cellular apoptosis susceptibility protein that plays important roles in mediating cell apoptosis induced by various cytotoxic toxins and chemotherapeutic drugs. Our studies showed that CAS over-expression increased p53 accumulation and apoptosis induced by 5-fluorouracil, doxorubicin, cisplatin, and tamoxifen in HT-29 cancer cells. A method based on coexpression of CAS with green fluorescence protein (GFP) was developed for high-sensitivity anticancer drug screening. Cancer cells transfected with CAS- and GFP-expressing vectors or the control and GFP expressing vectors were grown on 96-well microplates, treated with compounds to be screened, and detected with a microplate fluorescence reader. GFP fluorescence decreased following cancer cell death induced by the anticancer compounds. CAS transfection enhanced the cytotoxicities of anticancer compounds and therefore increased the decline in GFP fluorescence. Thus, anticancer compounds could be identified more sensitively. Our study indicates that CAS is an important p53 and apoptosis regulator and may be used for high-throughput anticancer drug screening as well as cytotoxic toxin assays. PMID- 20020939 TI - Retraction. PMID- 20020940 TI - Markers of long-term exposure to organophosphorus pesticides in farmers who work in viniculture and tobacco production in Turkey. AB - ABSTRACT The first aim of this study was to evaluate the degree of organophosphorus (OP) pesticides' exposure in viniculture and tobacco production workers via physical examination (neurology and general health aspect) and analysis of paraoxonase (PON1) and butyrylcholinesterase (BuChE) activities. The second aim was to investigate if PON1 polymorphism plays any role in long-term OP exposure. A total of 93 farmers who work as applicators in agriculture were studied. The data were evaluated according to agricultural type, and although the total exposure time was similar in both areas, BuChE and PON1 activities of farmers who work in tobacco production were lower. Overall, BuChE and PON1 activities showed a depletion in the farmer group compared to age-matched controls. When the farmers were categorized according to the number of their symptoms, the BuChE activities of farmers who had two or more symptoms were found to be depleted (n = 43, 2948 +/- 756) compared to farmers who had one or no symptoms (n = 37, 3356 +/- 659). Allele frequencies of patients and controls for positions 55 and 192 were similar to Turkish population data and there was no association between the allele polymorphism and symptoms/signs of long-term exposure. Our results indicate that there is an important inhibition of PON1 activity in chronic OP poisoning, and this together with BuChE activity might well be used as a reliable index of chronic exposure to OP. PMID- 20020941 TI - A simple method for accurate endotracheal placement of an intubation tube in Guinea pigs to assess lung injury following chemical exposure. AB - ABSTRACT Guinea pigs are considered as the animal model of choice for toxicology and medical countermeasure studies against chemical warfare agents (CWAs) and toxic organophosphate pesticides because of the low levels of carboxylesterase compared to rats and mice. However, it is difficult to intubate guinea pigs without damaging the larynx to perform CWA inhalation experiments. We describe an easy technique of intubation of guinea pigs for accurate endotracheal placement of the intubation tube. The technique involves a speculum made by cutting the medium-size ear speculum in the midline leaving behind the intact circular connector to the otoscope. Guinea pigs were anesthetized with Telazol/meditomidine, the tongue was pulled using blunt forceps, and an otoscope attached with the specially prepared speculum was inserted gently. Insertion of the speculum raises the epiglottis and restrains the movements of vocal cord, which allows smooth insertion of the metal stylet-reinforced intubation tube. Accurate endotracheal placement of the intubation tube was achieved by measuring the length from the tracheal bifurcation to vocal cord and vocal cord to the upper front teeth. The average length of the trachea in guinea pigs (275 +/- 25 g) was 5.5 +/- 0.2 cm and the distance from the vocal cord to the front teeth was typically 3 cm. Coinciding an intubation tube marked at 6 cm with the upper front teeth accurately places the intubation tube 2.5 cm above the tracheal bifurcation. This simple method of intubation does not disturb the natural flora of the mouth and causes minimum laryngeal damage. It is rapid and reliable, and will be very valuable in inhalation exposure to chemical/biological warfare agents or toxic chemicals to assess respiratory toxicity and develop medical countermeasures. PMID- 20020942 TI - Effect of chronic iron ingestion on the development of brush border enzymes in rat intestine. AB - ABSTRACT Iron is an essential element for biological systems. There is increasing evidence that excess iron supplementation results in the deposition of iron in the duodenum and enhances mucosal injury and cell proliferation in the colon and cecum. In the present study we examined whether chronic exposure to high levels of iron fortification affects the functional integrity of the small intestine, especially the activities of various brush border enzymes. Wistar rats were fed iron 29 mg/kg body weight (or 6.58 mg/kg Fe) daily in the form of FeSO(4).7H(2)O for 39 days. The activities of brush border alkaline phosphatase (AP) (p < 0.001), sucrase (p < 0.01), maltase (p < 0.05), lactase (p < 0.05), and trehalase (p < 0.001) were reduced in purified membranes in iron-fed animals compared to controls. However, the activities of leucine amino peptidase (LAP) and gamma glutamyl transpeptidase (gamma-GTP) were unaffected under these conditions. Analysis of alkaline phosphatase activity across the crypt-villus unit revealed a significant decrease (p < 0.05) all across the crypt-villus length, while sucrase activity was reduced (p < 0.01) only in the midvillus axis in iron-exposed animals. Kinetic studies showed a decrease in V(max) of AP from 1.11 to 0.83 units/mg protein and for sucrase from 0.77 to 0.43 units/mg protein in iron-fed rats, with no change in the apparent K(m) of the enzymes (AP, 8 mM; sucrase, 10 mM). Western blot analysis corroborated these findings. These results indicate that chronic iron exposure alters the activities of brush border enzymes, resulting in intestinal dysfunctions. PMID- 20020943 TI - New K-Oximes (K-27 and K-48) in Comparison with Obidoxime (LuH-6), HI-6, Trimedoxime (TMB-4), and Pralidoxime (2-PAM): Survival in Rats Exposed IP to the Organophosphate Paraoxon. AB - ABSTRACT Oximes are cholinesterase reactivators used in organophosphorus compound poisoning. The purpose of the study was to compare the protective effect of the K oximes (K-27 and K-48) in male rats with that of obidoxime (LuH-6), trimedoxime (TMB-4), and HI-6, using paraoxon (POX) as a cholinesterase inhibitor. Pralidoxime (2-PAM) was also retested. Seven groups of six rats each were used. Group 1 (G(1)) received 1 mumol/rat POX ( approximately LD(75)), the other groups (G(2-7)) received 1 mumol/rat POX + one of the six reactivators. The animals were monitored for 48 h and time of mortality was recorded. The procedure was repeated seven times. Subsequently, experiments as described were repeated using 10 and 15 mumol/rat POX. Mortality data were compared and hazards ratios (relative risks) ranked with the Cox proportional hazards model using the POX dose and group (reactivator) as time-independent covariables. K-27 followed by K-48 were the most potent reactivators. K-27 was statistically significantly superior to all other reactivators except K-48. The relative risk of death estimated by Cox analysis in K-27- and K-48-treated animals when compared with untreated animals, adjusted for the POX dose, was 0.22 (95% confidence interval [CI], 0.15 to 0.31) and 0.26 (95% CI, 0.18 to 0.37), respectively. We concluded that in the animal model used K-27 and K-48 are superior to older oximes in their ability to protect from paraoxon effects. They should be tested further using methyl- and propyl organophosphates as toxic agents. PMID- 20020944 TI - Evaluation of the Function of the Granzyme-Mediated Antiviral System through a Mathematical Model. AB - ABSTRACT Granzymes (gzms), the major components of cytolytic lymphocyte granules, are predominantly associated with immune regulatory processes against primary viral infections in vertebrates. By releasing into the infected cell cytosol, they initiate a rapid apoptotic process to eliminate infected cells. However, the kinetics and control mechanisms of gzms in antiviral processes have not yet been fully described. Based on earlier experimental data of ectromelia virus infection, we developed a mathematical model to evaluate the function of the gzm mediated antiviral system. The results of simulation runs for the infections corresponded well to the experimental data, suggesting that the computational methods could be applied in researching such field. Furthermore, by altering the parameters of the model and stability analysis, we found that for viruses with various virulence, gzm-mediated defense systems cannot completely suppress the first infections in most cases, unless the virulence is limited in a narrow range. Depending on the differences of viral virulence, gzms cause oscillations of viral level in different patterns. By adding secondary immune response to the model, we found that the oscillation evoked by the gzm-mediated antiviral system has crucial significance. Consequently, we bring forward a hypothesis that most of these oscillations may allow ancillary immune responses to effectively suppress a virus as it oscillates through a minimum level. PMID- 20020945 TI - Tumor Promotion and Oxidative Stress in Ferric Nitrilotriacetate-Mediated Renal Carcinogenesis: Protection by Adhatoda vasica. AB - ABSTRACT In the present study, we report the chemopreventive effects of Adhatoda vasica against ferric nitrilotriacetate (Fe-NTA)-induced renal oxidative stress, hyperproliferative response, and two-stage renal carcinogenesis. Fe-NTA (9 mg Fe/kg body weight, intraperitoneally) enhances renal lipid peroxidation, xanthine oxidase (XO), and hydrogen peroxide (H(2)O(2)) generation with concomitant reduction in renal glutathione content (GSH), antioxidant enzymes, and phase II metabolizing enzymes. It induces blood urea nitrogen, serum creatinine, ornithine decarboxylase (ODC) activity, and [(3)H] thymidine incorporation into renal DNA. It also enhances DEN (N-diethylnitrosamine)-initiated renal carcinogenesis by increasing the percentage incidences of kidney tumors. Pretreatment of rats orally with A. vasica (50 and 100 mg/kg body weight) resulted in a significant decrease in lipid peroxidation, H(2)O(2) generation, xanthine oxidase (XO), blood urea nitrogen, serum creatinine, renal ODC activity, DNA synthesis (p < 0.001), and incidence of tumors. Renal GSH (p < 0.01), glutathione-metabolizing enzymes (p < 0.001), and antioxidant enzymes were also recovered significantly (p < 0.001). Thus, our results show that A. vasica may meet the criteria demanded from a chemopreventive agent and in a rodent system it can reduce hyperproliferative response toxicity and carcinogenic activity of Fe-NTA. PMID- 20020946 TI - Hepatic neutrophil infiltration in the pathogenesis of alcohol-induced liver injury. AB - ABSTRACT The histopathologic evidence of the presence of neutrophils within the liver parenchyma is a prominent feature of alcoholic hepatitis in both experimental animals and chronic human alcoholics. However, the precise mechanisms by which neutrophils infiltrate the liver and cause liver injury still remain to be fully elucidated. For neutrophils to infiltrate the liver, they have to undergo systemic activation (priming) by proinflammatory cytokines, chemokines, complement factors, and other biologically active molecules (e.g., platelet-activating factor). Neutrophils accumulated in the hepatic microvasculature (sinusoids and postsinusoidal venules) can extravasate (transmigrate) into the hepatic parenchyma if they receive appropriate signals from previously sensitized or distressed cells. Transmigration can be mediated by a chemokine gradient established toward the hepatic parenchyma and generally involves the interaction between adhesion molecules on neutrophils (beta(2) integrins) and on endothelial cells (intercellular adhesion molecules [ICAM-1]). Following transmigration, neutrophils adhere to sensitized hepatocytes through their beta(2) integrins and ICAM-1 expressed on hepatocytes and mediate killing of hepatocytes mostly by oxidant stress and proteases. These neutrophilic events during chronic alcohol ingestion, based mostly on experiments with rodent models, will be emphasized in this review. PMID- 20020947 TI - Mass Spectrometry and Integrated Virus Detection System Characterization of MS2 Bacteriophage. AB - ABSTRACT In this study, we demonstrate the effect of sample matrix composition of MS2 virus on its characterization by ESI-MS and IVDS. MS2 samples grown and purified using various techniques showed different responses on ESI-MS than that on IVDS. The LC-MS of the specific biomarker of MS2 bacteriophage from an infected Escherichia coli sample was characterized by the presence of E. coli proteins. The significant impact of sample matrix was observed upon identification of MS2 using a database search. Infected E. coli with MS2 showed a matching score indifferent from uninfected ones. Only purified MS2, using CsCl and analyzed by LS-MS, showed a positive match using the database search. However, the variation in MS2 sample matrix had no effect on the deification of MS2. PMID- 20020948 TI - The Toxicity of Soman in the African Green Monkey (Chlorocebus aethiops). AB - ABSTRACT This study determines soman toxicity in African green monkeys (Chlorocebus aethiops) and is the first step in exploring the suitability of this species as a model for nerve agent studies. Male African green monkeys were surgically implanted with telemetry devices to monitor electroencephalographic (EEG) and electrocardiographic (ECG) activity. Blood was taken at various times to measure whole blood acetylcholinesterase (AChE) activity and cardiac troponin I (cTnI). Blood AChE activity relative to baseline was 0.0% to 2.5% 6 h after soman exposure and recovered to 31.9% to 72.0% by 30 days after exposure. The 6 h postexposure cTnI levels varied from 0.64 to 6.55 ng/mL, suggesting cardiac damage. Soman was prepared in saline to a concentration of 100 mug/mL. Using an up-down design for small samples, subjects were exposed to 5.01, 6.31, or 7.94 mug/kg soman IM. The first subject was given 5.01 mug/kg soman IM and survived. Three subjects received 6.31 mug/kg soman IM and survived. Three subjects received 7.94 mug/kg soman IM and died within 25 min, 26 min, or 6 h. In all subjects, toxic signs of muscle fasciculation, tremors, chewing, and profuse salivation developed within 2 to 7 min. Tonic-clonic motor convulsions and EEG seizure began between 2 and 18 min after tremor onset. The 48 h IM LD50 of soman in saline in the African green monkey was calculated to be 7.15 mug/kg. The signs and speed of soman intoxication in African green monkeys were consistent with those described in rhesus, cynomolguscynomolgus, and baboons. PMID- 20020949 TI - The Relationship Between Lipid Peroxidation and LDL Desialylation in Experimental Atherosclerosis. AB - ABSTRACT High serum total cholesterol concentration has been strongly connected with atherosclerosis in numerous studies. Being the main carrier of cholesterol in blood, low-density lipoprotein (LDL) is also the principal lipoprotein causing atherosclerosis. Sialic acids are a family of amino sugars that are commonly found as terminal oligosaccharide residues on glycoproteins and are sialylated on their apolipoprotein and glycolipid constituents. In several studies, it was demonstrated that LDL has a 2.5- to 5-fold lower content of sialic acid in patients with coronary artery disease compared with healthy subjects. The role of oxidatively modified LDL in the pathogenesis has been well documented. These studies have focused on modifications in the lipid and protein parts of LDL. But recently, desialylated LDL and its relation with the oxidation mechanisms have received attention in the pathogenesis of atherosclerosis and coronary artery disease (CAD). From these points, we have performed atheroma plaques in an experimental atherosclerosis model with rabbits and examined the LDL and plasma sialic acid and thiobarbituric acid reactive substance (TBARS) levels in the same model. We also have determined serum sialidase enzyme activities relevant with these parameters. LDL sialic acid levels were significantly decreased in the progression of the atherosclerosis (by the 30th, 60th, and 90th days). LDL and plasma TBARS levels and plasma sialidase enzyme activities were significantly elevated by the same time periods. In conclusion, serum sialidase enzyme may play an important role in the desialylation mechanism, and reactive oxygen substance (ROS) may affect this reaction. PMID- 20020950 TI - Role of vitamin b(6) deficiency in the nitrogen balance of streptozotocin diabetic rats. AB - ABSTRACT Male Wistar rats with streptozotocin-induced diabetes received a control diet and a pyridoxine-deficient diet. The animals were divided at random into four groups: control rats (CR), control diabetic rats (CDR), diabetic rats receiving a pyridoxine-free diet (DRB6), and diabetic rats receiving saline solution and no insulin treatment (DRSS). The experiment lasted 45 days. During the first 15 days the animals were observed for the development of diabetes and during the remaining 30 days they received the respective diets. The absence of vitamin B(6) did not influence the glycemia levels at the end of the experiment or the weight evolution of the animals. The rats that did not receive pyridoxine (DRB6) only showed a reduction in GPT activity (17.79 U/mL) compared to the other groups. The DRB6 group presented a significantly lower (p <0.05) nitrogen balance during each period (2.38 +/- 0.44 g N/7 days) compared to the CDR group (3.28 +/- 0.56 g N/7 days). The DRSS group presented similar or significantly higher values (2.81 +/- 0.77 g N/7 days) compared to the CDR group. Pyridoxine-deficient diabetic rats treated with insulin suffered important changes in the utilization of dietary proteins, as observed by nitrogen balance and enzyme activity studies. PMID- 20020951 TI - Beneficial effect of phosphodiesterase-5 inhibitor in experimental inflammatory bowel disease; molecular evidence for involvement of oxidative stress. AB - ABSTRACT Inflammatory bowel disease (IBD) is a common and chronic gastrointestinal disorder characterized by intestinal inflammation and mucosal tissue damage. Reactive oxygen metabolites (ROMs) play a pathogenic role in IBD. We aimed to examine the protective effect of sildenafil, a cGMP phosphodiesterase inhibitor, in the experimental mouse model of IBD. Intrarectal instillation of acetic acid was used to induce IBD. Prednisolone was used as the standard drug for comparison. Sildenafil was used at doses of 0.75, 1.5, and 3 mg/kg. Biochemicals and macroscopic and microscopic examinations of colonic tissue were performed. Results indicated that activity of myeloperoxidase (MPO) and lipid peroxidation product (TBARS) markers of oxidative stress are increased in acetic acid-treated groups and are recovered by sildenafil pretreatment and prednisolone. Sildenafil- (1.5 and 3 mg/kg) and prednisolone-treated groups showed significantly lower score values of macroscopic and microscopic characters when compared to the acetic acid-treated group. The beneficial effect of sildenafil (3 mg/kg) was comparable to that of prednisolone. It is concluded that sildenafil is helpful in the management of IBD, which is presumably related to its strong antioxidative stress potential mediated through enhanced cGMP. Results of proper clinical trials will determine the possible efficacy of phosphodiesterase-5 inhibitors in human IBD. PMID- 20020952 TI - Modulation by N-acetylcysteine of lead-induced alterations in rat brain: reduced glutathione levels and morphology. AB - ABSTRACT The present study pertains to the modulator action of N-acetylcysteine (NAC) on glutathione (GSH) status in lead-exposed brain regions of the rat. The effect of lead at a dose of 20 mg/kg body weight/day was studied on the cerebral cortex and cerebellum region of rat brain. The results showed a significant decline in the reduced glutathione level in the cerebellar and cerebral tissue homogenates. Histological analysis of the different brain regions also revealed a marked deterioration in the organization of the pyramidal cellular layer of the cerebrum and the Purkinje's cellular layer of the cerebellum. The animals that underwent lead treatment when administrated with N-acetylcysteine at a dose of 160 mg/kg body weight/day showed a significant recovery of the GSH level in the brain region, while it reached almost the normal level in the cerebellum. NAC treatment also brought about appreciable improvement in the histoarchitecture of the cellular layers, which showed clearly that NAC may play a very useful role in arresting the neurotoxicological damage of lead in cerebral and cerebellar brain regions. PMID- 20020953 TI - Methods and measurements for estimating human dermal uptake of volatile organic compounds and for deriving dermal permeability coefficients. AB - ABSTRACT Water contamination of public drinking and recreational waters with volatile organic compounds (VOCs) is widespread, resulting in human exposure through multiple routes. Although dermal absorption is of considerable importance, there is great uncertainty in the dermal permeability coefficient (K(P)) for many VOCs due to the methods by which they are derived. We present a human in vivo experimental approach for the measurement of VOC dermal uptake from water contaminated at environmentally relevant concentrations and for determination of K(P). Dermal permeability was estimated from 11 adult subjects following immersion of their hand and forearm into a sealed 4.8-L Plexiglas cylinder containing 100 mug/L each of chloroform, 1,1,1-trichloroethane (111 TCA), and toluene and 400 mug/L of methyl tertiary butyl ether (MTBE) in water. Uptake was determined by measuring the decrease in VOC water concentration during the exposure. A control glass arm accounted for nondermal losses. The concentration of VOCs was determined by solid-phase microextraction (SPME) and analysis with gas chromatography/mass spectrometry. For the eight male and three female adult subjects, on average there was a dermal uptake-attributed 13.5%, 14.9%, 20.8%, and 7.3% decrease in the concentration after 1 h exposure for chloroform, 111-TCA, toluene, and MTBE, respectively. The resulting mean K(P) (standard deviation) was estimated to be 0.166 (0.108), 0.167 (0.107), 0.250 (0.064), and 0.109 (0.157) cm/h for the respective analytes. The experimental K(P) values presented here exceed the previously published model-estimated K(P) values by factors ranging from 6 to 57, suggesting that the published model estimated K(P) values may underestimate actual VOC dermal absorption from water. PMID- 20020954 TI - Attenuation of Acetaminophen-Induced Hepatotoxicity In Vivo and In Vitro by a 43 kD Protein Isolated from the Herb Cajanus indicus L. AB - ABSTRACT The aim of this study was to evaluate the hepatoprotective role of a 43 kD protein (Hp-P) isolated from the leaves of Cajanus indicus L. against acetaminophen (APAP)-induced toxicity in mouse liver and in isolated hepatocytes. The hepatotoxicity of APAP and the hepatoprotective activity of Hp-P in vivo were determined by measuring the liver-specific serum marker enzymes alanine amino transferase (ALT) and alkaline phosphatase (ALP) in murine sera and observing the histological changes in the mice liver treated with the protein before and after (2 mg/kg body weight for 5 days) APAP (at a dose of 300 mg/kg body weight for 2 days) administration. The cell viability, LDH leakage, GSH level, and lipid peroxidation were measured in isolated hepatocytes to evaluate the cytotoxic effect of APAP and the protective role of Hp-P in vitro. Experimental results showed that APAP induced hepatotoxicity in vivo as revealed from the changes in serum-specific marker enzyme levels and histology of liver. It also induced cytotoxicity in hepatocytes as observed from the changes in cell viability and LDH leakage. Pretreatment with Hp-P prevented the APAP-induced elevation of ALT and ALP in murine sera. In addition, posttreatment with Hp-P significantly altered most of the changes induced by APAP. Although some natural recovery has been observed in toxin controls, the Hp-P-induced recovering process is more rapid than the natural ones. In histological studies, less centrilobular necrosis was found in the liver treated with Hp-P before and after APAP intoxication compared to the liver treated with APAP alone. Radical scavenging experiment showed that Hp-P scavenges DPPH radicals directly. Studies also showed that APAP induced reduced cell viability and cellular LDH leakage could be prevented by the combinatorial effect of Hp-P. Besides, treatment of hepatocytes with Hp-P and APAP together maintained the normal GSH level. APAP-induced enhanced lipid peroxidation was also decreased when cells were treated with APAP and Hp-P together. Hp-P alone, on the other hand, did not induce any alterations of the studied parameters. Results of this study have been compared with a known antioxidant, alpha-tocopherol. Data from both the in vivo (before and after APAP administration) and in vitro studies suggest that Hp-P has potent hepato- and cytoprotective properties against APAP-induced toxicity. PMID- 20020955 TI - In vivo Alterations in Glutathione-Related Processes, Lipid Peroxidation, and Cholinesterase Enzyme Activities in the Liver of Diazinon-Exposed Oreochromis niloticus. AB - ABSTRACT Although its usage is partially banned in developed countries, organophosphate (OP) pesticide diazinon finds extensive agricultural application in our country (Turkey). This study was conducted to evaluate the effects of diazinon on total glutathione (tGSH), GSH-related enzymes, cholinesterase (ChE) enzyme activities, and lipid peroxidation in the liver of Oreochromis niloticus, a freshwater fish, as a model organism. Fish were exposed to 0.1, 1, and 2 mg/L sublethal concentrations of diazinon for 1, 7, 15, and 30 days. Total GSH levels, GSH-related enzyme and ChE-specific activities, and malondialdehyde (MDA) levels were analyzed using spectrophotometric methods. tGSH levels are decreased at 1 day, while they were increased in the long-term period. GSH-related enzyme activities are affected by diazinon exposure, except glutathione reductase (GR; EC 1.6.2.4). Diazinon displayed an oxidative stress-inducing potential and it increased lipid peroxidation. Similar inhibition levels were observed in acetylcholinesterase (AChE; EC 3.1.1.7) and butyrylcholinesterase (BChE; EC 3.1.1.8.) enzyme activities, and these inhibitions were not dose dependent. ChE inhibition-related oxidative stress was observed using its correlation with elevated tGSH levels and increased glutathione S-transferase (GST; EC 2.5.1.18) enzyme activities; that reflects the diazinon-induced oxidative stress in the liver of O. niloticus. According to the results of the present study, tGSH level and GST-specific activity are suitable for reflecting the toxic effects of diazinon in fish. PMID- 20020956 TI - Microcalorimetric Investigation of the Toxic Effect of Iron Species on Escherichia coli. AB - ABSTRACT A microcalorimetric technique based on bacterial heat output was explored to evaluate the toxic effect of iron species on Escherichia coli. Power time curves of the growth metabolism of E. coli and the effect of different iron species on it were studied using the TAM III multichannel microcalorimetric system, isothermal mode, at 37 degrees C. The differences in shape of the power time curves and the thermodynamic and kinetic characteristics of E. coli growth have been compared. The thermodynamic parameters, that is, growth rate constant (k), inhibitory ratio (I), half-inhibitory concentration (IC(50)), P(max), and Q(total), have been calculated. The experimental results reveal that the sequence of antibiotic activity of the different iron species (three forms) on E. coli growth is Fe(3+) (ferric citrate) > Fe(2+) (ferrous chloride) > Fe(3+) (ferric chloride). These results are important to further studies of the physiology and pharmacology of iron species as antibacterial agents. PMID- 20020957 TI - Effects of bacterial toxins on endothelial tight junction in vitro: a mechanism based investigation. AB - ABSTRACT Lipopolysaccharide (LPS) and lipoteichoic acid (LTA), principal cell wall components of Gram-negative and Gram-positive bacteria, respectively, play a central role in altering the blood-brain barrier and facilitate bacterial infection of the host brain. Despite the significance of bacterial toxins in disease pathogenesis, mechanisms by which toxins impair the barrier are not yet known. This study, using an in vitro cell culture model, showed that LPS and LTA interacted with the endothelial cells and disrupted the tight junction between the cells that increased the barrier's permeability. Both toxins increased inducible nitric oxide synthase (iNOS) mRNA that is indicative of an increase in intracellular NO release, disrupted architecture of the tight junction proteins, suppressed zonula occludens-1 (ZO-1) and occludin (OCL) and junctional adhesive molecules (JAM) mRNA levels, and increased tumor necrosis factor alpha (TNFalpha) and interleukin-1 beta (IL-1beta) mRNA levels. Anti-CD14 antibodies blocked the increase in TNFalpha and IL-1beta mRNA levels but did not affect either changes in the tight junction or iNOS, ZO-1, OCL, and JAM mRNA levels in endothelial cells and astrocytes. Although both toxins did not cross the endothelial barrier, the abluminal neurons exhibited high inflammatory activity characterized by a sequential increase in TNFalpha, IL-1beta, external receptor kinase (ERK), and RelA-p50 that induced inflammation, followed by an increase in anti inflammatory/apoptotic factors including IL-10 and cysteine-aspartic acid protease-8 (CASPASE-8), which resolve inflammation and induce apoptosis. Anti CD14 antibodies in luminal buffer blocked the pro- and anti-inflammatory effects of the toxins in neurons. Thus, the CD14-TLR cascade that participates in the inflammatory effects of toxins may not participate in the toxin-induced barrier disruption in vitro. Since the toxins did not cross the endothelial barrier, induction of inflammation in neurons was due to a release of proinflammatory cytokines in the abluminal fluid. PMID- 20020958 TI - Effect of Ellagic Acid, a Plant Polyphenol, on Fibrotic Markers (MMPs and TIMPs) during Alcohol-Induced Hepatotoxicity. AB - ABSTRACT Alcoholic fibrosis and its end-stage cirrhosis occur when the rate of matrix synthesis exceeds matrix degradation. Hepatic fibroproliferation is associated with alterations of hepatic tissue inhibitors of matrix metalloproteinase (TIMPs) and matrix metalloproteinases (MMPs/matrixins) expressions. The alteration of hepatic matrixins and TIMPs expression to disease stage and inflammatory activity underlines their potential diagnostic markers in chronic liver disease. Ellagic acid (EA), a natural phenolic compound found in fruits and nuts, has potent antioxidant, anti-inflammatory, and anticancerous properties. The aim of our study was to gain further insight into the effect of EA on fibrotic markers (MMPs and TIMPs) during alcohol-induced tissue injury. To elucidate the effect on the MMPs/TIMPs balance by EA, gelatin zymography, multiwell zymography, succinylated gelatin assay, and ELISA technique (for TIMPs) were carried out. Coadministration of EA with alcohol decreased the expression of MMP-2 and -9 and TIMP-2 in a dose-dependent manner. These results suggest that EA at the dosage of 60 mg/kg body weight effectively decreased the expression pattern of fibrotic markers during alcohol-induced toxicity. Hence, it can be developed as an antifibrotic compound in near future. PMID- 20020959 TI - Glutathione as a mediator of the in vitro cytotoxicity of a green tea polyphenol extract. AB - ABSTRACT The 24-hr cytotoxicities of a green tea polyphenol (GTP) extract to cell lines derived from the human oral cavity were assessed using the neutral red (NR) assay. The sequence of sensitivity was carcinoma HSC-2 cells > immortalized gingival GT1 fibroblasts > normal gingival HGF-2 fibroblasts. The GTP extract generated hydrogen peroxide (H(2)O(2)) in cell culture medium and in phosphate buffer, albeit to a lesser extent. A 3-hr exposure to the GTP extract lowered the intracellular glutathione (GSH) content of the HSC-2 cells, but stimulated that of the GT1 and HGF-2 fibroblasts. The cytotoxicity of a 4-hr exposure of the GTP extract to the HSC-2 and GT1, but not to the HGF-2, cells was lessened in the presence of 2.5 mM GSH. Conversely, a 0.5 hr preexposure to the glutathione depleter, 1-chloro-2-dinotrobenzene (CDNB) at 25 muM, potentiated the 24-hr cytotoxicity of the GTP extract to the HSC-2 and GT1, but not to the HGF-2, cells. Using a cell-free system, it was shown that the GTP extract quickly depleted GSH, with depletion greatly enhanced at an alkaline pH, thus, correlating with the enhanced generation of H(2)O(2) by the GTP extract observed at alkaline pH. Apparently, a mode of cytotoxicity of the GTP extract, in particular to the carcinoma HSC-2 cells, was to induce oxidative stress, as noted by the generation of H(2)O(2), the depletion of intracellular GSH, the protection afforded by extracellular GSH, and cell hypersensitivity after pretreatment with CDNB. PMID- 20020960 TI - Role of glutathione in detoxification of copper and cadmium by yeast cells having different abilities to express cup1 protein. AB - ABSTRACT Although copper is an essential metal and cadmium is an environmental pollutant, both are toxic when present in excess. Metallothionein and glutathione are two of the key components that participate in detoxification of copper and cadmium. In the present study the role of glutathione in resistance to copper and cadmium was investigated with the yeast Saccharomyces cerevisiae. The yeast cells used in this study have different abilities to produce glutathione and Cup1 protein, the yeast metallothionein homolog encoded by CUP1 gene. It was demonstrated that Cup1 protein plays a dominant role in buffering excess copper, and yeast does not depend on glutathione to reduce copper toxicity whether it possesses single or multiple copies of CUP1. In fact, excess copper can cause glutathione oxidation and depletion and damage the glutathione system. On the other hand, it was indicated that Cup1 protein is an important cadmium detoxifying component, and the glutathione system can positively respond to cadmium. In yeast containing single or multiple copies of CUP1, glutathione is an indispensable line of defense against cadmium. Yeast having glutathione and no Cup1 protein is not able to grow in medium containing excess copper, but can tolerate higher concentrations of cadmium. In addition, it was found that yeast, independent of glutathione, can efficiently remove excess copper, whereas it cannot promptly eliminate accumulated cadmium regardless of having glutathione or not. PMID- 20020961 TI - Nonylphenol Causes Decrease in Antioxidant Enzyme Activities, Increase in O(2)(-) Content, and Alteration in Ultrastructures of FG Cells, a Flounder (Paralichthys olivaceus) Gill Cell Line. AB - ABSTRACT The findings in this study were that the exposure of FG cells to nonylphenol (NP) caused decreases in the cell growth rate, antioxidant enzyme activities, and ATP content; an increase in intracellular O(2)(-) content; and alteration of the organelles such as mitochondria and the RER. It is proposed that the reduction in antioxidant enzyme activities leads to accumulation of O(2)(-) content, which in turn results in the damage of mitochondrial infrastructures and a decline in energy production, which then causes the ultrastructural alterations in the cells. In addition, we show that FG cells are a good candidate system for evaluation of NP acute toxicities. PMID- 20020962 TI - Genotoxicity of DNA Intercalating Anticancer Drugs: Pyrimido[4(I),5(I):4,5] thieno(2,3-b)quinolines on Somatic and Germinal Cells. AB - ABSTRACT The genotoxic effect of the anticancer drugs Oxo-PTQ, Morpholino-PTQ, and Chloro-PTQ were studied in mice bone marrow by means of chromosome aberrations and micronucleus assay. These drugs, at dose levels of 21.42, 28.57, and 35.71 mg/kg b.w., respectively, were given to mice in a single application via the intraperitoneal route. Marrow was collected at 24, 48, and 72 h after the application. The chromosome aberrations and micronucleus assay were done according to standard procedures. These compounds gave rise to an increase in the number of micronuclei in a dose-dependent manner. The number of micronucleated polychromatic erythrocytes showed a maximum at 24 h, and there was partial recovery at 72 h. Chromosome aberrations increased significantly as compared to normal controls when treated with Oxo-PTQ followed by Morpholino-PTQ at 21.42 mg/kg b.w. (48 h). Statistically significant sperm abnormalities also revealed the genotoxic potency of these drugs. Our results suggest that the Pyrimido[4(I),5(I):4,5]thieno(2,3-b)quinoline drugs owe at least some of their cytotoxicity to their genotoxic effects, which seem to be mediated through interaction with topoisomerase II. PMID- 20020963 TI - Anticlastogenic Activity of Aqueous Extract of Agaricus blazei in Drug Metabolizing Cells (HTCs) During Cell Cycle. AB - ABSTRACT The mushroom Agaricus blazei has been extensively investigated because of evidence of its antimutagenic, antitumor, and anticarcinogenic activities. This study investigated the clastogenic and/or anticlastogenic activity of aqueous extract of Agaricus blazei (10% w/v) in drug-metabolizing rat hepatoma tissue cells (HTCs), with continuous treatment and treatment during different phases of the cell cycle. DNA damage was induced utilizing two direct-acting agents-methyl methane sulfonate and ethyl methane sulfonate-and two indirect acting agents-2-aminoanthracene and cyclophosphamide. The aqueous extract of A. blazei with either continuous treatment or treatment during different phases of the cell cycle showed clastogenic activity. The results with continuous treatment showed that A. blazei does not protect against DNA damage-inducing agents that are direct acting. Meanwhile, when combined with indirect-acting agents, a protective effect was demonstrated. A protective effect was also found during different phases of the cell cycle when cells were treated with indirect-acting agents. The protective effects against indirect-acting agents (continuous treatment and during the different phases of the cell cycle) suggest that A. blazei may provide some health benefits to the public when used as a functional food. PMID- 20020964 TI - Influence of umbelliferone on glycoprotein components in diabetic rats. AB - ABSTRACT Umbelliferone (7-hydroxycoumarin), a derivative of coumarin, is benzopyrone in nature, and it is present in the fruits of golden apple (Aegle marmelos Correa) and bitter orange (Citrus aurantium). The present study was designed with the objective of evaluating the effect of umbelliferone (UMB) on glycemic control and glycoprotein components in the plasma and tissues (liver, kidney, heart, and brain) of streptozotocin (STZ)-induced diabetic rats. Adult male albino rats of Wistar strain, weighing 180 to 200 g, were induced diabetes by administration of STZ (40 mg/kg b.w.) intraperitoneally. The normal and diabetic rats were treated with UMB dissolved in 10% dimethyl sulfoxide (DMSO) for 45 days. In our study, diabetic rats had decreased plasma insulin and elevated blood glucose, sialic acid, total hoxoses, fucose, and hexosamines in the plasma and tissues. Treatment with UMB brought the levels of glucose, sialic acid, total hoxoses, fucose, and hexosamines to near-normal level. Thus, our results show that treatment with UMB has improved glycemic control and thereby reduced the formation of glycoprotein components. This leads to the normalization of circulatory and tissue sialic acid, fucose, hexoses, and hexosamines in STZ diabetic rats, which reflects a protective effect of UMB from the risk of diabetic complications. PMID- 20020965 TI - Efficacy of 2-PAM, TMB-4, HI-6, and Toxogonin in Reactivation of Immobilized Acetylcholinesterase Inhibited by Compound R-33. AB - ABSTRACT Efficacy of reactivators 2-PAM, TMB-4, HI-6, and toxogonin in reactivation of acetylcholinesterase inhibited by compound R-33 has been studied. The study was performed with acetylcholinesterase immobilized and stabilized on a cotton fabric and with an indicator paper with acetylthiocholine iodide and 5,5' dithiobis(2-nitrobenzoic acid). The enzyme activity was determined by measurement of remission changes of the fabric surface, caused by reaction of the chromogenic reagent with products of enzymatic hydrolysis of the substrate. The most potent of the reactivators studied was 2-PAM at concentrations 0.5 mg.cm(-3) and 0.1 mg.cm(-3). Effectivity of this reactivator was over 20%. At these concentrations, reactivator HI-6 was less effective. Efficacy higher than that of 2-PAM was observed only after 20 min at concentrations 0.05 mg.cm(-3) and 0.01 mg.cm(-3). For TMB-4 and toxogonin the reactivation values were about 10% after 10 min, whereas after 15 or 20 min these reactivators were ineffective. PMID- 20020966 TI - Assessment of mercury concentrations in mouse brain using different routes of administration and different tissue preparations. AB - ABSTRACT The objective of this study was to determine the best method for preparing brain tissue for mercury analysis from mice exposed to methylmercury either through subcutaneous (SC) injection or through ingestion. C57BL/6 J mice at postnatal day 29 were exposed to 0.0 or 5.0 mg/kg methylmercuric chloride (MMC) given SC or through food containing MMC. Eighteen mice received vehicle (sodium bicarbonate; SC) and 18 additional mice received 5.0 mg/kg MMC (SC). Whole brain tissue was prepared using one of four tissue preparation methods: rapid freezing, saline perfusion, 4% paraformaldehyde perfusion fixation, or 4% paraformaldehyde immersion fixation. Brains from vehicle-treated mice exhibited minimal levels of mercury (0.0007 to 0.0018 ppm) in all preparation methods. Mercury content in rapidly frozen control brains differed statistically from immersion-fixed control brain tissue. There was no significant difference in mercury content from mice given 5.0 mg/kg MMC (SC) in all preparation methods (0.2660 to 0.3650 ppm). Additional mice were divided into groups of six mice each: single oral dose of 5.0 mg/kg MMC; total oral dose of 5.0 mg/kg MMC divided into five doses; and vehicle only. Forebrain (0.3243 ppm) and hindbrain (0.1908 ppm) mercury content in MMC-treated mice given multiple doses was 10 times higher than in brain tissue from mice given a single 5.0 mg/kg dose. Brain mercury content following administration of 5.0 mg/kg MMC via the oral route (0.5354 ppm) differed statistically from the SC route (0.3430 ppm). In conclusion, different tissue preparation methods do not significantly affect brain mercury content, but route of administration and dosing regimen can influence total brain mercury content. PMID- 20020967 TI - Vanadium-induced testicular toxicity and its prevention by oral supplementation of zinc sulphate. AB - ABSTRACT Transition metal vanadium has been shown to modulate the cellular redox potential and catalyze the generation of reactive oxygen intermediates. Since free radical production and lipid peroxidation are potentially important mediators in testicular physiology and pathophysiology, the present study was conducted to elucidate the vanadium-induced oxidative damages in rat testis and the ameliorative role of zinc sulphate against such adverse effects of vanadium. Adult male rats were dosed for 26 days with daily intraperitoneal injection of 0.4 mg V/kg body weight as sodium metavanadate. One group of rats was treated with zinc sulphate orally simultaneously with vanadium for 26 days, while the other group was treated with zinc sulphate alone. Changes in testicular and accessory sex organ weight, different varieties of germ cells at stage VII of spermatogenic cycle, epididymal sperm count, and enzymatic (Delta(5)3beta- HSD, 17beta- HSD, SOD, catalase), lipid peroxidation, and hormonal milieu were monitored. Vanadium treatment resulted in a significant increase in the testicular lipid peroxidation and caused a marked inhibition in the activities of antioxidant and steroidogenic enzymes. Histopathological examination revealed inhibition of spermatogenesis and the preferential loss of maturing and elongated spermatids. However, coadministration of zinc sulphate to vanadium-treated animals resulted in normalizing these parameters appreciably, emphasizing the therapeutic potentials of zinc. Taken together, the results suggest that an increase in free radical formation relative to loss of the antioxidant defense system during vanadium exposure may render testis more susceptible to oxidative damage, leading to their functional inactivation. However, zinc sulphate supplementation can be an effective antidote in the treatment of vanadium poisoning. PMID- 20020968 TI - Therapeutic Efficacy and Safety of Undenatured Type II Collagen Singly or in Combination with Glucosamine and Chondroitin in Arthritic Dogs. AB - ABSTRACT This investigation was undertaken to evaluate the therapeutic efficacy and safety of glycosylated undenatured type II collagen (UC-II) alone or in combination with glucosamine HCl and chondroitin sulfate in arthritic dogs. Twenty dogs divided into four groups (n = 5) were daily treated orally for 120 days: group I, placebo; group II, 10 mg UC-II; group III, 2,000 mg glucosamine + 1,600 mg chondroitin; group IV, UC-II (10 mg) + glucosamine (2,000 mg) + chondroitin (1,600 mg), followed by a 30-day withdrawal period. On a monthly basis, dogs were examined for overall pain, pain upon limb manipulation, and exercise-associated lameness. Serum samples were analyzed for markers of liver function (ALT and bilirubin) and renal function (BUN and creatinine). Body weight was also measured at a monthly interval. Dogs in group I exhibited no change in arthritic conditions. Dogs receiving UC-II alone showed significant reductions in overall pain within 30 days (33%) and pain upon limb manipulation and exercise associated lameness after 60 days (66% and 44%, respectively) of treatment. Maximum reductions in pain were noted after 120 days of treatment (overall pain reduction, 62%; pain reduction upon limb manipulation, 91%; and reduction in exercise-associated lameness, 78%). The overall activity of the dogs in the UC-II supplemented with glucosamine and chondroitin group (group IV) was significantly better than the glucosamine + chondroitin-supplemented group (group III). Glucosamine and chondroitin alleviated some pain, but in combination with UC-II (group IV) provided significant reductions in overall pain (57%), pain upon limb manipulation (53%), and exercise-associated lameness (53%). Following withdrawal of supplements, all dogs (groups II to IV) experienced a relapse of pain. None of the dogs in any groups showed any adverse effects or change in liver or kidney function markers or body weight. Data of this placebo-controlled study demonstrate that daily treatment of arthritic dogs with UC-II alone or in combination with glucosamine and chondroitin markedly alleviates arthritic associated pain, and these supplements are well tolerated as no side effects were noted. PMID- 20020969 TI - Chemopreventive effects of nonsteroidal anti-inflammatory drugs on 1,2 dimethylhydrazine-induced colon carcinogenesis in rats. AB - ABSTRACT The anticancer efficacy of two different classes of NSAIDs, the nonspecific cyclooxygenase (COX) inhibitor aspirin and the specific COX-2 inhibitor celecoxib, was examined at their therapeutic anti-inflammatory doses during 1,2-dimethylhydrazine (DMH)-induced colon carcinogenesis in a rat model. Eight to 10-week-old male rats of Sprague strain were divided into four groups. While group 1 served as control and received the vehicle of the drugs, groups 2, 3, and 4 were administered freshly prepared DMH in 1 mM EDTA saline (pH 7.0) (30 mg/kg body weight/week, subcutaneously). Groups 3 and 4 were also given a daily treatment of aspirin (60 mg/kg body weight, orally) and celecoxib (6 mg/kg body weight, orally), respectively, both prepared in carboxy-methyl cellulose. Animals were sacrificed at the end of 12 weeks and colons from different groups were subjected to macroscopic and histopathological studies, enzymatic activities of superoxide dismutase (SOD) and catalase (CAT), and determination of lipid peroxide level. The maximum number of raised mucosal lesions in proximal, middle, and distal regions of the colon was found in the DMH group alone, and the lowest number was found in the celecoxib-treated DMH group. Histological studies also showed the highest occurrence of dysplastic aberrant crypt foci (ACF) associated with enlarged lymphoid follicles in all the three portions of colon (i.e., proximal, middle, and distal). The aspirin-administered DMH group had lesser ACF in the proximal and middle portions and no ACF in the distal region. The celecoxib-administered DMH group showed no ACF in the middle region of the rat colon. DMH treatment induced lipid peroxidation and inhibited the activities of SOD and CAT. Both the aspirin- and celecoxib-treated DMH groups showed a marked lowering of the lipid peroxide level along with a significant enhancement of CAT activity when compared with the DMH-treated group. The results show that celecoxib was found to be more effective in reducing the ACF occurrence and aggregates of lymphoid tissue than the nonselective COX inhibitor aspirin, and suggests a possible chemoprevention modality in colon cancer. This may have important implications as COX-2 selective drugs at anti-inflammatory doses are better tolerated clinically than standard NSAIDs, thus making them potentially better chemopreventive agents in colon cancer. PMID- 20020970 TI - A case report of opium body packer; review of the treatment protocols and mechanisms of poisoning. AB - ABSTRACT Body packers are people who illegally carry drugs, mostly cocaine as well as opium and/or heroin, concealed within their bodies. The packets are inserted in the mouth, rectum, or vagina in order to get across borders without being detected. In this presentation we report a case of an opium body packer and review the available scientific literature by focusing on mechanisms of toxicity and treatment approach. The patient was a 35-year-old man who had lethargy, respiratory depression, tachycardia, normal blood pressure, hyperthermia, and pinpoint pupils on presentation. No past medical history was obtained and the only positive history was his travel from Afghanistan 2 days earlier, which he had given to emergency personnel before arriving at our hospital. Complete blood cells and kidney and liver tests were all in normal range. In the emergency department, the patient was treated with oxygen, naloxone, and hypertonic glucose. One dose of activated charcoal (1 g/kg) was administered orally. After intravenous injection of naloxone (4 mg), the lethargy, respiratory depression, and miosis were resolved. The patient was admitted to the intensive care unit and 90 min after admission, the patient redeveloped respiratory distress and lost consciousness. He was intubated and mechanically ventilated due to the suspicious of body packing. Plain abdominal x-ray showed multiple packets throughout the gastrointestinal tract; 81 packets were removed by surgery and three of them were left due to leaking. After removing the packets, the patient was treated conservatively. He suffered a pulmonary infection (aspiration pneumonia) and he regained consciousness after 4 days. Upon recovery the patient was seen by a psychiatrist prior to going to prison. Surgery is recommended for body packers who have significant signs or symptoms. PMID- 20020971 TI - Protection by pentoxifylline of diazinon-induced toxic stress in rat liver and muscle. AB - ABSTRACT The effects of diazinon, pentoxifylline, and their combination therapy on plasma glucose, the key enzymes of glycogenolysis and gluconeogenesis, and oxidative stress were studied in rat liver and muscle. Oxidative stress was determined by measuring the concentration of lipid peroxides and assessing total antioxidant capacity. Diazinon (60 mg/kg) and pentoxifylline (100 mg/kg) were administrated by gavage. Administration of diazinon increased blood glucose, hepatic glycogen phosphorylase (GP), and phosphoenol pyruvate carboxykinase (PEPCK) by 160.65%, 117.2%, and 93.5%, respectively, while it decreased plasma cholinesterase (ChE) by 53.82%. Diazinon-induced oxidative stress was demonstrated by decreased total antioxidant capacity and enhanced lipid peroxidation by 52.61% and 280% in liver and by 40.02% and 46.6% in muscle, respectively. Pentoxifylline increased plasma glucose, hepatic GP, and PEPCK by 98.65%, 60%, and 79.86%, respectively, while it did not change plasma ChE, liver and muscle lipid peroxides, and total antioxidant capacity. In combination therapy, pentoxifylline did not alter diazinon-induced change in muscle GP activity but restored a diazinon-induced increase in hepatic and muscle lipid peroxides by 39.18% and 42.35%, respectively. Pentoxifylline also recovered a diazinon-induced decrease in liver and muscle total antioxidant capacity and plasma ChE by 122.33%, 56.44%, and 115.62%, respectively. Pentoxifylline did not affect diazinon-induced hyperglycemia and increased hepatic GP and PEPCK or muscle GP activities. It is concluded that pentoxifylline is a good choice for the alleviation of acute toxic stress of diazinon in muscle and liver and ChE in plasma, while it is unable to recover diazinon-induced hyperglycemia. PMID- 20020972 TI - Ultrafine Quartz-Induced Damage in Human Lymphoblastoid Cells in vitro Using Three Genetic Damage End-Points. AB - ABSTRACT Respirable quartz is a potential human lung carcinogen. The mechanisms involved in this carcinogenesis, however, remain unclear, especially for the ultrafine particles (diameter <100 nm). The aim of the present study was to investigate the effects caused by ultrafine quartz (UF-quartz) in a human cell culture model. Genotoxicity and cytotoxicity induced by UF-quartz were investigated through the cytokinesis block micronucleus assay (CBMN), the Comet assay, the HPRT assay, the population growth assay, and the 3-(4, 5 dimethythiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. WIL2-NS cells were incubated for 10h with 0, 60, and 120 mug/mL UF-quartz. Significant decreases in percent of cell survival in the MTT assay were seen at higher doses, for example, 83%, and 64% relative survival at 60 mug/mL and 120 mug/mL, respectively. Only slight population regrowth was observed, with the population sizes recovering slightly by day 4 after quartz particles were removed. A significant increase in the frequency of micronucleated binucleated cells (MNed BNCs) was seen with 120 mug/mL quartz, from approximately 5 in 1000 BNCs in controls to 12 in 1000 BNCs. A significant reduction in the nuclear division index was observed by the CBMN assay, indicating inhibition of cell division by high-dose UF-quartz. A dose-dependent increase in induced HPRT-gene locus mutant frequency with increasing dose of UF-quartz was observed by the HPRT assay. No significant difference was found in DNA strand breakage as detected by the Comet assay. Collective findings suggest that UF-quartz can cause cytotoxicity and genotoxicity to human lymphoblasts in this model system. PMID- 20020973 TI - Effect of hesperidin on nicotine toxicity and histopathological studies. AB - ABSTRACT The study was designed to evaluate the protective nature of the bioflavonoid hesperidin on nicotine-induced toxicity in male Wistar rats. Lung toxicity was induced by subcutaneous injection of nicotine at a dose of 2.5 mg/kg of body weight (5 days a week for 22 weeks). The nicotine-treated rats showed a significant increase in the marker enzymes, cholesterol, triglycerides, phospholipids, and free fatty acids. There was a significant protection on hesperidin administration at a dose of 25 mg/kg body weight in nicotine-treated rats. This finding is supported by histopathological studies of liver, lung, and kidney. The results of the present study suggest that hesperidin showed protective effects against nicotine-induced toxicity. PMID- 20020974 TI - Effect of melatonin on N-nitrosodiethylamine-induced hepatocarcinogenesis in rats with reference to biochemical circadian rhythms. AB - ABSTRACT Tumors and tumor-bearing hosts exhibit markedly altered circadian rhythms, which serve as markers in the early diagnosis and prognosis of cancer. Our study presents the effect of melatonin on circadian rhythms of lipid peroxides and antioxidants in N-nitrosodiethylamine (NDEA)-induced hepatocarcinogenesis. The circadian rhythm characteristics (acrophase, amplitude, and mesor) of thiobarbituric acid reactive substances (TBARS), superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx), and reduced glutathione (GSH) were markedly altered in NDEA-treated rats. Melatonin administration caused a significant increase in the amplitude and mesor values of antioxidants and a significant decrease in the mesor values of TBARS. Further delays in acrophase in NDEA-treated rats were reversed by melatonin administration. In conclusion, melatonin may exert its chemopreventive effect by its role as an antioxidant as well as by altering the circadian rhythm characteristics. PMID- 20020975 TI - Adaptation of an in vitro phospholipidosis assay to an automated image analysis system. AB - ABSTRACT Phospholipidosis is the excessive intralysosomal accumulation of phospholipids and is induced in humans and animals by the chronic administration of cationic amphiphilic drugs. To identify compounds that may induce phospholipidosis early in the discovery process, we have developed a predictive fluorescent cell-based assay amenable to automated high content screening using the 2-(4,4-difluoro-5-methyl-4-bora-3a,4a-diaza-s-indacene-3-dodecanoyl)-1 hexadecanoyl-sn-glycero-3-phosphocholine (ss-BODIPY C(12)-HPC) dye and primary rat hepatocytes. ss-BODIPY C(12)-HPC localized to lysosomes that accumulate phospholipids and not to lipid droplets, indicating the selectivity for phospholipid-containing granules. Accumulation of ss-BODIPY C(12)-HPC was monitored in primary rat hepatocytes plated onto 96-well plates and 24 h after exposure to increasing concentrations of 13 drugs known to induce phospholipidosis and four negative compounds. Fluorescent images were captured and analyzed using the Discovery-1 automated cellular imaging system. Eleven out of the 12 selected positive compounds and all negative compounds were properly assigned as positive and negative inducers of phospholipidosis, respectively, indicating the high degree of sensitivity and specificity of this assay. The ability of ss-BODIPY C(12)-HPC to detect and quantify phospholipidosis is similar to that of the well-established probe, N-(7-nitrobenz-2-oxa-1,3-diazol-4-yl) dipalmitoylphosphatidylethanolamine (NBD-PE). PMID- 20020976 TI - Comparative measurement of cyanide and paraquat mitochondrial toxicity using two different mitochondrial toxicity assays. AB - ABSTRACT Cyanide (KCN) and paraquat (PQ) are very toxic to mitochondria. In this study the toxicity of KCN and PQ in the isolated rat liver mitochondria was determined using the MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] assay and JG-B (Janus green B) assay by multiwell scanning spectrophotometry. JG-B was used not only for the vital staining of mitochondria, but also for the mitochondrial viability assay and was compared to the MTT assay. The rat liver mitochondria were first isolated by centrifuge in a mixture of 0.25 M saccharose solution and 0.05 M Tris buffer. Various concentrations of paraquat (0.001 to 100 mM) and KCN (0.0001 to 100 M) on the mitochondria isolated from the liver were investigated. The 50% lethal concentration of toxins were found for PQ (4.45 +/- 0.02, 4.96 +/- 0.01) and KCN (0.22 +/- 0.02, 0.49 +/- 0.02), as determined by these assays ( JG-B and MTT, respectively ). Significant correlations were also observed among the two methods with a 95% coefficient interval (r(2) = 0.84, p < 0.001; r(2) = 0.91, p < 0.001; PQ and KCN, respectively). These results suggest that both methods are reliable and are comparable for determining the mitochondrial assay. It is concluded that the JG-B assay may be preferable to the MTT assay because of its simplicity, low cost, sensitivity, and objectivity; in addition, this method is not time dependent. PMID- 20020977 TI - Protective effect of black tea polyphenols against 7,12-dimethylbenz[a]anthracene induced genotoxicity and oxidative stress during hamster buccal pouch carcinogenesis. AB - ABSTRACT This study was designed to evaluate the protective effect of black tea polyphenols (Polyphenon B) against genotoxicity and oxidative stress during 7,12 dimethylbenz[a]anthracene (DMBA)-induced hamster buccal pouch (HBP) carcinogenesis. Since the bone marrow reflects hematopoietic stress caused by tumor cells, we used the frequency of micronuclei, the extent of lipid peroxidation, and the status of antioxidants in the bone marrow plasma as intermediate biomarkers of oxidative stress. All the hamsters painted with DMBA alone for 14 weeks developed buccal pouch carcinomas with a 75.4% increase in the incidence of bone marrow micronuclei as compared to untreated control (group 4). This was accompanied by an increase in lipid peroxidation as evidenced by the formation of thiobarbituric acid reactive substances (TBARS) and lipid hydroperoxides (LOOH) (61.3% and 17.8%, respectively) and a decrease in the ratio of reduced glutathione (GSH) to oxidized glutathione (GSSG) and the activities of GSH-dependent enzymes glutathione peroxidase (GPx) and glutathione S-transferase (GST) by 32.6%, 35.9%, and 62.8%, respectively, as compared to untreated control. Dietary administration of Polyphenon B significantly suppressed DMBA-induced HBP carcinomas by 20% and reduced the frequency of bone marrow micronuclei by 57.3% and TBARS and LOOH by 38.8% and 25.8%, respectively, compared to group 1 animals with significant elevations in the GSH:GSSG ratio (16.0%) and activities of GPx (29.8%) and GST (53.8%). Our results reveal that dietary supplementation of Polyphenon B exerts protection against DMBA-induced genotoxicity and oxidative stress by augmenting bone marrow antioxidant defense mechanisms. PMID- 20020978 TI - Differential phosphorylation of MAPK isoforms in keratinocyte cell line by contact allergens and irritant. AB - ABSTRACT Mitogen-activated protein kinase (MAPK) is an important mediator of cellular signaling pathways. In an attempt to investigate the role of these kinases in contact allergen/irritant-induced signaling cascades, we have utilized keratinocyte (A431 cell line) as antigen presenting cells (APCs). This group has previously reported the applicability of A431 as an APC. The role of these kinases in the elicitation pathways of allergic contact dermatitis was investigated using interleukin-12p40 (IL-12p40) as a marker. Inhibitors of the isoforms of MAPK reduced the allergen-induced IL-12 release in the A431 cell line. This result suggests a possible role of the isoforms of MAPK in the development of contact allergy. PMID- 20020979 TI - Protective Effects of alpha-Tocopherol and N-Acetyl-Cysteine on Diazinon-Induced Oxidative Stress and Acetylcholinesterase Inhibition in Rats. AB - ABSTRACT Diazinon, an organophosphate (OP) insecticide, is widely used in agriculture and domestically. Reactive oxygen species (ROS) caused by OPs are involved in the toxicity of various pesticides. The aim of the present study was to analyze the role of diazinon in inducing oxidative stress in adult male Wistar rats and to evaluate the possible protective effects of alpha-tocopherol (TPH) and the glutathione prodrug N-acetyl-cysteine (NAC) after 4 weeks of exposure to a sublethal dose of diazinon. TPH (10 mg/kg/day), NAC (160 mg/kg/day), diazinon (25 mg/kg/day), a combination of NAC (160 mg/kg/day) and diazinon (25 mg/kg/day), and a combination of TPH (10 mg/kg/day) and diazinon (25 mg/kg/day) were given to rats orally via gavage for 4 weeks. The thiobarbituric acid reactive substances (TBARS) marker of lipid peroxides levels, total thiol molecules, and total antioxidant capacity of plasma were all analyzed as biomarkers of oxidative stress. In addition, the acetylcholinesterase (AChE) activity was measured as a biomarker of toxicity. The results from this study well indicate diazinon-induced oxidative stress demonstrated by enhanced TBARS, decreased total thiol molecules, and total antioxidant capacity. In addition, AChE activity was inhibited as a marker of OP toxicity. Data show the protective roles of TPH and NAC in reducing the diazinon-induced oxidative stress. Interestingly, both TPH and NAC recovered diazinon-induced AChE inhibition. It is concluded that supplementation with TPH and NAC can reduce toxicity of OP in human exposure. PMID- 20020980 TI - Effect of Selenium and alpha-Tocopherol on the Antioxidant Defense System of Goat Erythrocytes and the Hemic System. AB - ABSTRACT The effect of sodium selenite, alpha-tocopherol, and their combination was studied in the hemic system of goat. Incubation of goat blood with selenium and vitamin E for 24 h resulted in a decrease in hemoglobin concentrations in Se and vitamin E-treated groups, whereas no change in the osmotic fragility was observed. In plasma the levels of albumin did not show any significant change in all the treatment groups. The acute phase protein (total protein) was found to be increased in the group treated with Se at a concentration of 10 mug, and ceruloplasmin in the plasma was significantly increased in the group supplemented with Se at a concentration of 20 mug. The catalase was found to be increased in both the Se treatment groups and the group that received vitamin E at a concentration of 10 mug. However, the effects on lipid peroxidation, acetylcholinesterase, and antioxidant enzymes such as superoxide dismutase are not significant. The sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) pattern of the proteins did not show any alteration. Also, no effect was observed in the membrane permeability and physicochemical alteration of erythrocytes in the presence of Se and vitamin E. PMID- 20020981 TI - Errata. PMID- 20020982 TI - The Genotoxicity of Priority Polycyclic Aromatic Hydrocarbons (PAHs) Containing Sludge Samples. AB - ABSTRACT In this research work we developed in vitro tests utilizing mammalian cell cultures, which can rapidly assess effect of exposure of oily sludge-derived chemicals on human and ecological health. Many of these are hazardous to health and environment due to their toxicity and/or accumulation potential in sediments as well as in organisms. Petroleum refinery and petrochemical industry-derived oily sludges contain toxic polycyclic aromatic hydrocarbons (PAHs), some of which are lipophilic in nature. Risk assessment of environmental samples suffers from inadequate availability of toxicity data, lack of knowledge about behavior of genotoxic substances in complex matrices, paucity of information on synergistic and antagonistic interactions of mixture of components, etc.; the literature describing the behavior of genotoxic substances in complex mixtures is sparse and sometimes contradictory. The present study aims at assessing the genotoxic potential of oily sludges collected from an integrated petroleum refinery and petrochemical industry located in the southwestern part of India and a petrochemical industry located in the western part of India using a battery of genotoxicity assays such as DNA damage/strand break, chromosomal aberration, p(53) protein induction, and apoptosis in CHO-K1 cell culture system. Exposure with different dose levels of sludge extracts (25, 50, 100 muL) in CHO-K1 cells could cause statistically significant level of (P < 0.001) DNA damage, chromosomal aberration, p(53) protein induction, and apoptosis in comparison to negative control treatment groups, and the genotoxicity was attributed to PAHs present in the sludge as identified by GC-MS. This implies that the sludges are genotoxic in nature in mammalian cells tested, and the exposure to these may pose a potential genotoxic risk to human beings. PMID- 20020983 TI - Factors affecting the accuracy of bisphenol a and bisphenol a-monoglucuronide estimates in Mammalian tissues and urine samples. AB - ABSTRACT Bisphenol A (BPA) (CAS Number 80-05-7; EINECS Number 201-245-8) is used in the production of plastics having food contact applications. Some biomonitoring studies have reported free BPA in blood or urine of humans. Since complete first-pass metabolism of orally administered BPA to BPA-monoglucuronide (BPA-G) occurs in humans, the presence of free BPA in human specimens raises questions as to the origin and/or possible sources of the free BPA. We hypothesized that BPA-G instability during specimen collection and analysis contributes to the presence of free BPA in the biological samples. Investigation of the in vitro hydrolysis of BPA-G in blood plasma, tissue homogenates, and diluted urine from laboratory rats and in aqueous/organic solutions commonly used for extraction in BPA analyses lent support to the hypothesis of BPA-G instability as a possible source of free BPA determinations in the biological specimens. Hydrolysis of BPA-G occurred at neutral pH and room temperature in diluted urine and in rat placental or fetal tissue homogenates at room temperature. Hydrolysis of BPA-G in aqueous/organic solutions began within minutes at pH 2 and 80 degrees C. BPA-G was degraded to an unidentified compound in a urine/water mixture or when stored in a 25/75 mixture of urine/acetonitrile at pH 9 at either 22 or 80 degrees C. Based upon these experiments, it was concluded that methods demonstrating BPA-G stability or accounting for its instability during analysis are warranted in studies designed to measure free BPA in biological specimens. PMID- 20020984 TI - Influence of diallyl disulphide on temporal patterns of circulatory lipid peroxidation products and antioxidants in N-nitrosodiethylamine-induced hepatocarcinogenesis in rats. AB - ABSTRACT Investigations on the effects of diallyl disulphide (60 mg/kg body weight) orally administered (thrice a week) on the temporal patterns of thiobarbituric acid reactive substances (TBARS), superoxide dismutase (SOD), catalase, and reduced glutathione (GSH) during N-Nitrosodiethylamine (NDEA) induced hepatocarcinogenesis were performed in rats. The acrophase of TBARS was found to be delayed and of antioxidants was found to be advanced in NDEA-treated rats. The increase in mesor of TBARS, decrease in mesor of antioxidants, and altered amplitude and acrophase indicated the negative imbalance of oxidant and antioxidant occurring during carcinogenesis. Oral treatment of diallyl disulphide (DADS) results in the resynchronization of the altered rhythms of TBARS and other antioxidants. Although NDEA has no known significant effects on the suprachaismatic nucleus (SCN), from the present results it could be hypothesized that it would influence the peripheral oscillator systems, such as liver, possibly by modulating secretion of the various hormones and growth factors during hepatocarcinogenesis. PMID- 20020985 TI - Role of an aminothiazole derivative on ethanol-induced toxicity. AB - ABSTRACT The protective effect of dendrodoine analog (DA) [4-amino-5-benzoyl-2-(4 methoxy phenylamino) thiazole] at three doses (5, 10, and 15 mg/kg body weight) was investigated on ethanol-induced hyperlipidemia. Hepatotoxicity was induced by administering 7.9 g ethanol/kg body weight for 45 days by intragastric intubation. Our results showed increased activity of aspartate transaminase (AST), alkaline phosphatase (ALP), and gamma glutamyl transferase (GGT) and increased levels of cholesterol, triglycerides, and phospholipids in the plasma of alcohol-given group when compared with normal control group. The levels of tissue (liver and kidney) cholesterol and triglycerides were increased significantly in alcohol control rats when compared with normal control rats. The levels of phospholipids decreased significantly in the liver and kidney of alcohol control rats when compared with normal control rats. The activity of phospholipase A and phospholipase C increased significantly in the liver of alcohol control rats when compared with normal control rats. Intragastric administration of DA at 10 mg/kg body weight effectively lowered the activity of hepatic marker enzymes (GGT, AST, and ALP), phospholipase A, and phospholipase C, and decreased the levels of plasma and tissue lipids. The level of tissue phospholipids increased significantly when DA was administered at a dose of 10 mg/kg body weight along with alcohol when compared with alcohol control group. Thus, we propose that DA exerts a hepatoprotective effect by modulating liver marker enzymes and lipid levels at a dosage of 10 mg/kg body weight. PMID- 20020986 TI - Protein Isolate from the Herb Phyllanthus niruri Modulates Carbon Tetrachloride Induced Cytotoxicity in Hepatocytes. AB - ABSTRACT Phyllanthus niruri is a well-known hepatoprotective herbal plant. In the present study, hepatoprotective potential of the protein isolate of P. niruri was investigated against carbon tetrachloride (CCl(4))-induced hepatoxicity in vitro. Isolated hepatocytes were treated with CCl(4) and also separately with various concentrations of the protein isolate of P. niruri along with CCl(4). Levels of different marker enzymes related to hepatic integrity and different antioxidant enzymes as well as lipid peroxidation products in hepatocytes were measured in normal, control (toxicity induced), and protein isolate-treated cells. Administration of CCl(4) increased the leakage of glutamate pyruvate transaminase (GPT) by four fold and lactate dehydrogenase (LDH) by 84% in cell suspension, along with increased lipid peroxidation (114%), and reduced the levels of antioxidant enzymes superoxide dismutase (SOD) and catalase (CAT) to almost 30% and 37% of the normal values, respectively. Treatment with the protein isolate of P. niruri significantly altered these changes. GPT value almost came down to normal levels and LDH value was reduced to 32% of normal values. Depletion of SOD and CAT activities were restored significantly to 75% and 87% of normal values, respectively. Lipid peroxidation was also reduced significantly. In the DPPH free radical scavenging activity, the protein isolate was also able to quench the free radical. Results suggest that the protein isolate of P. niruri protects hepatocytes against CCl(4)-induced oxidative damage and may be used as an effective cytoprotector against CCl(4)-induced hepatotoxicity. PMID- 20020987 TI - Alterations in the lipid profile and membrane dynamics of rat intestinal brush border membrane induced by different classes of nonsteroidal anti-inflammatory drugs. AB - ABSTRACT Nonsteroidal anti-inflammatory drugs (NSAIDs) are known to cause gastrointestinal damage. New anti-inflammatory drugs have been developed in an attempt to improve their gastrointestinal side effect profile. Therefore, the objective of the present study was to compare the effect of three different NSAIDs, aspirin, nimesulide, and celecoxib, on the lipid profile and dynamics of rat intestinal brush border membranes (BBMs). Female Wistar rats were divided into four different groups viz: group I (control), group II (aspirin treated), group III (nimesulide treated), and group IV (celecoxib treated). Groups II, III, and IV received the corresponding drugs orally at a dose of 40 mg/kg body weight dissolved in water, while the control group received the vehicle only. After 28 days, all the treatment groups showed modification in the lipid profile of intestinal BBM as compared to control. Alterations in the intestinal membrane dynamics by fluidity studies showed a considerable increase, which correlated well with the changes in the lipid composition. It was suggested that NSAIDs such as aspirin, nimesulide, and celecoxib pose intestinal side effects due to initial changes in the composition and dynamics of the membranes. It was further concluded that newly discovered NSAIDs such as celecoxib have better safety profiles because of the less intense membrane effects, but studies are still required to comment decisively on the suitability of various NSAIDs depending upon their cyclooxygenase enzyme specificity. PMID- 20020988 TI - Protective effect of ursolic Acid against myocardial ischemia induced by isoproterenol in rats. AB - ABSTRACT Sustained high levels of circulating catecholamines may induce cardiotoxicity through oxidative mechanisms. Isoproterenol is a synthetic catecholamine with increasing attention owing to this application in cardiology. The aim of the present study was to investigate the cardioprotective effects of ursolic acid against isoproterenol-induced myocardial ischemia. Normal Wistar strain rats were pretreated with UA (20, 40, and 60 mg/kg, s.c.) for 7 days and then intoxicated with isoproterenol (ISO, 85 mg/kg, s.c. for 2 consecutive days). Hearts were excised from the experimental animals and assessed for the activities of cardiac markers [alanine aminotransferase (ALT), aspartate aminotransferase (AST), lactate dehydrogenase (LDH), and creatine phosphokinase (CPK)], the levels of lipid peroxide products [thiobarbituric acid reactive substances (TBARS), lipid hydroperoxides (HPs), and conjugated dienes (CDs)], myeloperoxidase (MPO), lipid profiles [total cholesterol (TC), free cholesterol, ester cholesterol, triglycerides (TG), free fatty acids (FFAs), and phospholipids (PLs)], and membrane-bound enzymes (total ATPase, Na(+)K(+)ATPase, Ca(2+)ATPase, and Mg(2+)ATPase). In ISO-treated group, shrinkage of cardiac markers and elevated lipid peroxidation with compromised lipid profiles in the heart where accompanied by the decreased activities of membrane-bound enzymes. The prior administration of UA significantly (p < 0.05) prevented the isoproterenol-induced alterations and restored the enzymes to near normal. These findings indicate the cardioprotective activities of UA during isoproterenol-induced myocardial ischemia. PMID- 20020989 TI - Protective role of pongamia pinnata leaf extract on tissue antioxidant status and lipid peroxidation in ammonium chloride-induced hyperammonemic rats. AB - The present investigation was designed to study the protective role of Pongamia pinnata (an indigenous plant used in Ayurvedic medicine in India) leaf extract on oxidative stress during ammonium chloride-induced hyperammonemia by measuring the extent of oxidative damage as well as antioxidant status. Ethanolic extract of Pongamia pinnata (PPEt) leaves was administered orally (300 mg/kg body weight) and the effects of PPEt on the levels of thiobarbituric acid reactive substances (TBARS), hydroperoxides (HP), conjugated diene (CD), superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx), and reduced glutathione (GSH) were studied in liver and kidney of ammonium chloride-induced hyperammonemic rats. On treatment with PPEt, a significant reduction in the levels of TBARS, HP, and CD and a significant increase in the levels of SOD, CAT, GPx, and GSH in liver and kidney of ammonium chloride-induced hyperammonemic rats were observed, which clearly shows the antioxidant property of PPEt. The study of induction of the antioxidant status is considered to be a reliable marker for evaluating the antiperoxidative effect of the medicinal plant. Our present findings show the protective role of PPEt against lipid peroxidation and suggest that PPEt possesses antioxidant potential that may be used for therapeutic purposes. The exact mechanism has to be still investigated and the isolation of active constituents is required. PMID- 20020990 TI - Detecting Bacteria by Direct Counting of Structural Protein Units by IVDS and Mass Spectrometry. AB - This report explores the direct counting of "hair-like" struc-tures specific for Gram-positive bacteria. Indications show that these structures are intact after removal from the cell and are sufficiently different from species to species of bacteria to give an indication of bacteria type if not actual identification. Their detection would represent a new approach to bacteria detection and identification. This report documents the detection of the bacterial structures using the physical nanometer counting methodology in the Integrated Virus Detection System (IVDS) and electrospray ionization-mass spectrometry. PMID- 20020991 TI - Effect of Chlorpyrifos on the Hematology and Phagocytic Activity of Nile Tilapia Cells (Oreochromis niloticus). AB - Chlorpyrifos is an organophosphorous insecticide widely used in agriculture and domestic activities. However, little is known about the effect of this pesticide on the immune system of fish and other alterations of its physiological system. The aims of the present study were the evaluation of the LC(50) (lethal concentration(50)) and the potential toxicity of this substance on Nile tilapia (Oreochromis nilotucus), as well as its effect on some hematological values and phagocytic functions of this fish. Results obtained showed that chlorpyrifos does not have any effect on the following parameters: number of red blood cells, hemoglobin, hematocrit, mean corpuscular volume (MCV), mean corpuscular hemoglobin (MCH), and mean corpuscular hemoglobin concentration (MCHC). However, the phagocytic index and the percentage of phagocytic active cells were significantly affected, which could indicate that phagocytic parameters from Nile tilapia are more sensitive than hematological ones to assess the effect of acute intoxication with chlorpyrifos. PMID- 20020992 TI - Changes of serum erythropoietin during Cisplatin- or 5-Fluorouracil-induced anemia in rats. AB - The aim of the present work was to investigate changes of serum erythropoietin (Epo) during cisplatin (DDP)-or 5-Fluorouracil (5-Fu)-induced anemia in rats. Anemia was induced with single intravenous (IV) injection of DDP of 8 mg/kg body weight or 5-Fu of 150 mg/kg body weight. Serum Epo, hemoglobin (Hb), blood urea nitrogen (BUN) concentration, and reticulocyte (Ret) counts were measured 7, 21, 35, and 49 days after administration of the anticancer drugs. The changes of renal tissue were examined by light microscope. A single injection of 5-Fu decreased Ret counts and Hb concentration and increased serum Epo levels. No evident morphological abnormalities were found by light microscopy in 5-Fu treated rats. A single injection of DDP decreased Ret counts and Hb concentration and increased BUN. Serum Epo was decreased 7 days but was increased 21 days after DDP treatment; however, serum Epo elevation was modest. Pathological examination revealed that DDP caused significant renal damage in rats. These results suggest that in 5-Fu-induced anemia, the elevated serum Epo level was due to the feedback response to anemia. In DDP-induced anemia, the elevated serum Epo level was low in relation to the level of anemia, and DDP-induced nephrotoxicity might be the main cause of changes of serum Epo. PMID- 20020993 TI - Gadolinium Chloride Inhibits the Spontaneous Resolution of Fibrosis in CCL(4) Induced Cirrhosis. AB - Current evidence indicates that liver fibrosis is dynamic and can be bidirectional, involving phases of progression and regression, and that in addition to increased matrix synthesis, this pathological process involves major changes in the regulation of matrix degradation. There is also evidence that Kupffer cells participate in both fibrogenesis and fibrolysis. Therefore, the aim of the present work was to study the participation of Kupffer cells on the spontaneous resolution of hepatic fibrosis. Cirrhosis was produced by 3 months of chronic CCl(4) intoxication in male Wistar rats, and then CCl(4) was discontinued and two groups were formed: One group received gadolinium chloride (10 mg/kg, IP, daily) and the other received the vehicle (water) only for 2 months. Serum enzyme activities of alkaline phosphatase and alanine aminotransferase and liver lipid peroxidation increased by CCl(4) treatment but returned to normal by discontinuation of CCl(4). GSH, GSH/GSSG, and GSH+GSSG decreased significantly by CCl(4), but withdrawal of CCl(4) restored normal glutathione parameters. Fibrosis increased five-fold and glycogen decreased significantly by CCl(4) treatment, while discontinuation of CCl(4) reversed completely glycogen depletion and partially fibrosis. Gadolinium chloride showed effects only in the content of glycogen and collagen; the former was decreased further and the latter remained elevated despite discontinuation of the toxic agent. Persistent fibrosis induced by gadolinium chloride, a selective inhibitor of Kupffer cells, indicates that these cells play a pivotal role in fibrolysis. PMID- 20020994 TI - Mechanism of DNA Strand Cleavage Induced by Hexaaza Macrocyclic Nickel (II) Complex. AB - The hexaaza macrocyclic nickel(II) complex (Ni(II)L-1,8-Dihydroxyethyl 1,3,6,8,10,13-hexaazacyclotetradecane nickle(II) perchlorate monohydrate) was synthesized and purified. The electrochemical character of Ni(II)L was measured, and the interaction of Ni(II)L with calf thymus DNA (CT-DNA) was studied using electrochemical techniques, emission and viscometry, and circular dichroic spectral measurements. All of the experiments suggested that the complex interacted with DNA primarily by partial intercalation. The cyclic voltammetry (CV) showed that the currents of both the reduction peak and the oxidation peak decreased significantly in the presence of DNA, which indicated that Ni(II)L could interact with DNA. The fluorescence intensity of the DNA-ethidium bromide(EB) system decreased distinctly when Ni(II)L was added. The results indicated that Ni(II)L may be completed effectively with EB for the intercalative binding sites. The viscosity of DNA would be decreased slightly by the addition of the complex. Circular dichroic spectral studies revealed that B conformation of CT-DNA became more A-like in structure on interaction with the complex. Noticeably, the complex has been found to cleave plasmid pBR 322 by agarose gel electrophoresis and cleave CT-DNA by high-performance liquid chromatography (HPLC). PMID- 20020995 TI - In vivo and in vitro characterization of the biochemical and pathological changes induced by lionfish (pterios volitans) venom in mice. AB - Accidents caused by lionfish (Pterios volitans) envenomation are characterized by edema, intense pain, and necrosis at the site of sting. The mode of action and biochemistry of venoms are obviously complex and require a better knowledge and investigation to explore the toxic action and resulting biochemical changes. In the present study the LD(50) value of lionfish venom was found to be 42.5 mug/kg body weight (intraperitoneal injection) in Albino Swiss mice and was associated with reduced motor activity and asphyxiation followed by respiratory failure. The effect on vital organs revealed spongiosis in brain, vascular congestion in liver, cloudy swelling of renal tubules, congested blood vessels in renal tubules, and degeneration of myofibrils in heart. Whereas, the 10% of LD(50) (was 4.25 mug/kg b.w.), the sublethal dose showed reversible changes in the hematological (blood cell count, hematocrit, hemoglobin, mean corpuscular volume, mean corpuscular hemoglobin, and platelet count) parameters, serum enzymes (aspartate transaminase, alanine transaminase, alkaline phosphatase, lactate dehydrogenase, and creatinine phosphokinase), blood sugar, urea, creatinine, triglycerides, cholesterol, and total protein in mouse in vivo. The in vitro analysis of lionfish venom on mouse brain acetyl cholinesterase and Na(+), K(+), ATPase showed significant increased activity in a dose-dependent manner (10 to 40 mug). Moreover, the lionfish venom was observed to have a protease with a molecular weight of 45 kDa. Hence, the present study suggests the presence of bioactive proteins and peptides with excellent target specificity, which could be trapped for drug development in near future. PMID- 20020996 TI - Genomics and human health-an assessment of the state of the science. PMID- 20020997 TI - The evolution of gene expression studies in drug safety assessment. AB - Since the identification in the 1950s of deoxyribonucleic acid as the building block of life, the impact of molecular biology has been far-reaching. Understanding the processes of how DNA is replicated, transcribed into RNA and then translated into protein products has not only provided a fundamental knowledge of life but has also spawned a plethora of applications. Molecular biology has been high profile and widespread in research into the biology of disease and in drug discovery. It has additionally found application in understanding the adverse effects, or toxicity, of candidate drugs and how they interfere with biochemical and biological processes. In recent times the biggest impact of molecular biology in toxicology has been through the study of differential gene expression, largely as a result of the advent of genomics. This review seeks to describe how toxicogenomics strategies have been implemented and integrated into nonclinical studies of drug safety. PMID- 20020998 TI - Gaining Confidence on Molecular Classification through Consensus Modeling and Validation. AB - Current advances in genomics, proteomics, and metabonomics would result in a constellation of benefits in human health. Classification applying supervised learning methods to omics data as one of the molecular classification approaches has enjoyed its growing role in clinical application. However, the utility of a molecular classifier will not be fully appreciated unless its quality is carefully validated. A clinical omics data is usually noisy with the number of independent variables far more than the number of subjects and, possibly, with a skewed subject distribution. Given that, the consensus approach holds an advantage over a single classifier. Thus, the focus of this review is mainly placed on how validating a molecular classifier using Decision Forest (DF), a robust consensus approach. We recommended that a molecular classifier has to be assessed with respect to overall prediction accuracy, prediction confidence and chance correlation, which can be readily achieved in DF. The commonalities and differences between external validation and cross-validation are also discussed for perspective use of these methods to validate a DF classifier. In addition, the advantages of using consensus approaches for identification of potential biomarkers are also rationalized. Although specific DF examples are used in this review, the provided rationales and recommendations should be equally applicable to other consensus methods. PMID- 20020999 TI - Genomic approaches for investigating mechanisms of genotoxicity. AB - The current genetic toxicity testing battery enables an accurate and effective detection of genotoxicity associated with exposure to chemicals. However, the interpretation of the data in light of the relevant risk to humans is often difficult due to limited insight into underlying genotoxic mechanisms. Thus, the development of experimental approaches capable of differentiating genotoxic mechanisms is expected to facilitate risk assessment. Recent progress in science and technology has enabled the investigation of the stress response associated with chemical exposure at the genomic level. For instance, gene expression profile analysis, transcriptomics, was proposed as a tool for evaluating toxic mechanisms. In addition, the advancement in genetic tools has allowed the study of stress response on a functional level, functional genomics. This review will outline a number of recent developments in genomic analyses of genotoxic stress response and provide a perspective on their application in genetic toxicology. PMID- 20021000 TI - Toxicogenomics and cross-species biomarker discovery: applications in drug discovery and safety assessment. AB - Toxicogenomics has evolved into a useful technique for providing greater mechanistic insights into adverse effects that will spur the development of novel approaches for identifying and understanding toxicity issues. The ability to capture a snapshot of the transcriptome at any given time during the development of an adverse phenotype allows unprecedented molecular views into the dynamic physiological changes that are occurring on either time or dose continuum for a toxicology study of interest advancing our basic knowledge of adverse events, and providing the necessary scientific framework for developing new strategies and tools for safety assessment programs. The development of an effective subset of cost effective devices for identifying toxicity earlier in the drug development process will help identify the most promising candidate compounds to move forward leading to a reduction in compound attrition due to toxicity. In addition, there is a need in the pharmaceutical industry to develop safety and efficacy biomarkers that are relevant to multiple species such as rat, dog, and human. Genomics provides an opportunity to discover novel cross-species biomarkers for identifying phenotypes such as liver fibrosis, especially if the biomarkers are tissue specific secreted proteins that can be monitored in the serum. This review includes an example of how databases from multiple species, in this case rat and human tissues, can be utilized to identify candidate cross-species diagnostic markers of hepatitis and fibrosis. This study illustrates a genomic approach for identifying candidate cross-species biomarkers of cirrhosis/fibrosis for humans and rats, and a previously known biomarker of fibrosis (APOA1) and a novel candidate biomarker of fibrosis, FETUB were identified. As more "omic" databases are built, a reservoir of molecular information will become available for toxicologist to gain more extensive views on the physiological alterations induced by adverse events, which will inevitably lead to the development of better tools for predicting, identifying, categorizing, and determining cross species impact of the toxicity and ultimate provide a novel scientific scaffold for improving safety assessment protocols. PMID- 20021001 TI - Application of pharmacogenomics in clinical pharmacology. AB - Many factors can affect a patient's response to a drug. These include intrinsic factors such as age, gender, race/ethnicity, genetics, disease states, organ dysfunctions, and other physiological changes, including pregnancy, lactation, and extrinsic factors such as smoking, diet (food, juice, dietary supplements), and concomitant medications (ICH E5, 1998 and 2004). The interplay of genotypes of the enzymes, transporters and receptors, among other factors (such as concomitant medications and disease states), can affect the risk/benefit ratio for individual patients. This commentary discusses when the genomic information should be obtained during drug development and when it is to be assimilated into labeling and standards of care that can be used to "individualize" drug therapy and become one of the pillars of "personalized medicine." PMID- 20021003 TI - Exploring genotypic and phenotypic diversity of microbes using microarray approaches. AB - Application of genome-scale analysis like DNA microarray technology has revolutionized multiple scientific disciplines. Herein, a next generation of DNA microarrays, a DNA tiling approach that allows high throughput sampling of genomes with single-nucleotide precision, is described. As methods revealing a genomic scale examination of cellular phenotypes offer keen insights for genomic analyses, a high throughput system for whole cell phenotyping is similarly detailed. The merit of these technologies in discriminating pathogenic and commensal strains of microbes is emphasized using the microbe, Escherichia coli, as an example. Deployment of microarray strategies to assess closely-related microbial strains should help address diversity of organisms in their feral settings. PMID- 20021002 TI - "Omics" Technologies and the Immune System (a) , (b). AB - Recent advances in genomics-based identification of gene families and gene polymorphisms associated with immune system dysfunction have answered basic questions in immunology and have begun to move forward our understanding of immune-related disease processes. In toxicology, "omic" technologies have the potential to replace or supplement current immunotoxicological screening procedures, to provide insight into potential mode or mechanisms of action, and to provide data suitable for risk assessment. The application of omic technologies to the study of the immune system also has great potential to appreciably impact the diagnosis and treatment of immune-related diseases. This review focuses on the use of omic technologies in immunopharmacology and immunotoxicology, specifically considering the potential for these technologies to impact chemical hazard identification, risk characterization and risk assessment, and the development and application of novel therapeutics. The state of the science of omics technologies and the immune system is addressed in terms of a continuum of understanding of how omics technologies can and cannot yet be applied in the various aspects of immunopharmacology and immunotoxicology. Additionally, information gaps are identified that, once addressed, will move each area further down the continuum of understanding. PMID- 20021004 TI - DNA microarray technology in the evaluation of weight management potential of a novel calcium-potassium salt of (-)-hydroxycitric Acid. AB - Quality and quantity of diet and nutrients are key factors of human health and disease prevention. Molecular diagnostics and cellular signaling play a fundamental role in the usefulness of novel nutraceuticals and functional foods. Increasing knowledge of the genes and molecules involved in the development of obesity is creating new methods of obesity regulation. Traditional herbal medicines may have some potential in weight management. Botanical dietary supplements often contain complex mixtures of phytochemicals that have additive or synergistic interactions. Evidence from numerous human and animal dietary studies has demonstrated the potential therapeutic effects of traditional herbal medicines in controlling obesity. We analyzed the effects of low-dose oral administration of calcium-potassium salt of (-)-hydroxycitric acid (HCA-SX) on the body weight and abdominal fat transcriptome in rats. HCA-SX restricted body weight gain in rats and lowered abdominal fat leptin expression. High-density microarray analysis of 9960 genes and ESTs present in the fat tissue identified a small set of specific genes sensitive to dietary HCA-SX. Mitochondrial/nuclear proteins necessary for fundamental support of the tissue were not affected by HCA SX, further demonstrating its safety. Functional characterization of HCA-SX sensitive genes revealed that up-regulation of genes encoding serotonin receptors represents a distinct effect of HCA-SX on appetite suppression. PMID- 20021005 TI - Some major landmarks in the path from nuclein to human genome (1). AB - The completion of sequencing of the human genome as well as the genomes of other species is a spectacular achievement of 20th-century biology. It is appropriately regarded as a turning point in biology and medicine for the 21st century. Knowledge of the human genome will presumably help us understand the genetic instructions that make us human. By learning about the gene sequences, the functional dynamics of the genome as well as the individual genetic differences, scientists hope to understand the molecular basis of the normal state and the diseased state of life on one hand, and develop ways to individualize medicine and nutrition on the other hand. Thus, the science of genomics that has grown out of this genome sequencing effort is expected to revolutionize the future of biology itself. The present article is an attempt to briefly summarize some major landmarks in the path that began with the discovery of "nuclein" and led to the completion of the human genome sequencing. PMID- 20021006 TI - Methyl Parathion-Induced Changes in Free and Protein-Bound SH Levels in Rat Tissues. AB - The main objective of this study was to investigate the changes in free and protein-bound SH contents in methyl parathion-exposed rat tissues. The free and protein-bound SH levels are usually affected and depleted by oxidative stress inducing agents. Results would indicate if methyl parathion toxicity partly results from depletion of sulfhydryl content of tissues. Six-week-old male Wistar albino rats were used in this study. Following exposure to methyl parathion for 3 months, the liver, the brain, and the kidney tissues were removed from the rats. The free and protein-bound SH contents were determined in these tissues. In addition, plasma lactate dehydrogenase levels were determined. Our results showed that methyl parathion exposure significantly lowers the free and protein-bound SH levels in rat tissues. However, lactate dehydrogenase activity in the blood plasma did not display any differences compared to the control group. The free SH concentrations in the control rat liver, brain, and kidney tissues were 3.78 +/- 0.1 mumol/100 mg tissue, 1.56 +/- 0.08 mumol/100 mg tissue, and 2.16 +/- 0.08 mumol/100 mg tissue, respectively, whereas the free SH concentrations in rats exposed to methyl parathion were determined as 0.536 +/- 0.1 mumol/100 mg tissue in the liver, 1.06 +/- 0.1 mumol/100 mg tissue in the brain, and 0.108 +/- 0.03 mumol/100 mg tissue in the kidney. The protein-bound SH concentrations in the liver and in the kidney in rats exposed to methyl parathion displayed a significant decrease also. However, the protein-bound SH level in the brain did not change significantly. These results indicate that methyl parathion exposure partially depletes the free and protein-bound SH levels. Thus, it was concluded that methyl parathion toxicity may partly result from oxidative stress. PMID- 20021007 TI - Methanol intoxications in adana, Turkey. AB - Methanol is a highly toxic substance that is commonly used as an industrial solvent and antifreeze in automobiles. Methanol intoxication usually occurs following inhalation or topical exposure of spirit, homemade alcohol, or eau de cologne. The goal of this study was to examine methanol fatalities and methanol poisoning characteristics in our region and to contribute to form a database concerning methanol intoxication cases in our country. The records of the Morgue Department, Council of Forensic Medicine Adana Group Administration, were reviewed retrospectively for all methanol poisoning deaths for the period of January 1, 1997, to December 31, 2003. Age, gender, death places, methanol blood levels of the cases, and consumed products were recorded. Total number of deaths between 1997 and 2003 was 4725 and the number of deaths due to methanol poisoning was 41 (0.86%) during this period; 38 (92.68%) of the total methanol fatality cases were male and three (7.32%) female, with an age range of 16 to 54 years. PMID- 20021008 TI - A method for the analysis of tabun in multisol using gas chromatographic flame photometric detection. AB - Preparation and analysis of tabun (GA) solutions are necessary for the continued development of countermeasures to this nerve agent. GA solutions must be stable and compatible for use in the test systems chosen for study; however, GA is very unstable in saline solutions. In the past we have found GA in saline at 2 mg/mL to be stable for a month or less at -70 degrees C, whereas saline solutions of sarin (GB), soman (GD), and cyclosarin (GF) were stable for many months. Previous studies have shown that Multisol (48.5% H(2)O, 40% propylene glycol, 10% ethanol, and 1.5% benzyl alcohol) provides stable solutions of GA. We confirmed the stability of GA in Multisol with phosphorus nuclear magnetic resonance (P horizontal line NMR) and developed a method for the analysis of GA in Multisol using gas chromatographic flame photometric detection (GCFPD) in the phosphorus mode. The GC method used acetonitrile (CH(3)CN) for a dilution solvent because of its miscibility with GA in chloroform (CHCl(3)) standards and GA in Multisol samples at 1% (v/v). Furthermore, the dilutions with CH(3)CN made the phosphorus mode interference peak present in CHCl(3) analytically manageable, reduced the interferences of Multisol in the GC separation, and contributed to a safe and reliable analysis of GA at 20 mug/mL. We demonstrated the stability of GA in Multisol stored for more than a year at 70 degrees C. This method contributes a suitable technique for the preparation and analysis of reliable solutions of GA in nerve agent medical research and demonstrates the extended stability of GA in Multisol. PMID- 20021009 TI - Biochemical and histopathological evidences for beneficial effects of satureja khuzestanica jamzad essential oil on the mouse model of inflammatory bowel diseases. AB - The essential oil from Satureja Khuzestanica Jamzad (SKEO), an endemic plant from Iran, was evaluated for its activity against inflammatory bowel disease (IBD). SKEO was examined on the experimental mouse model of inflammatory bowel disease, which is acetic acid-induced colitis. Prednisolone was used as the standard drug for comparison. Biochemical, macroscopic, and microscopic examinations of colon were performed. Lipid peroxidation significantly increased in acetic acid-treated mice in comparison to the normal group (4.88 vs. 3.02 mumol/g) and was significantly restored by SKEO (500, 1000, 1500 ppm) and prednisolone treatment. The mean percentage of decreases of lipid peroxidation in SKEO (500, 1000, 1500 ppm)- and prednisolone-treated groups were 10.5, 28.5, 42.85, and 33.33 of control, respectively. The myeloperoxidase activity significantly increased in acetic acid-treated mice in comparison to the normal group (4.1 vs. 0.8 U/g) and significantly restored in SKEO (1000 and 1500 ppm)- and prednisolone-treated groups. The mean percentage of decreases of myeloperoxidase activity in SKEO (1000 and 1500 ppm)- and prednisolone-treated groups were 24.56, 50, and 52.63 of control, respectively. SKEO (1000 and 1500 ppm)- and prednisolone-treated groups showed significantly lower score values of macroscopic and microscopic characters when compared to the acetic acid-treated group. The beneficial effect of SKEO (1500 ppm) was comparable to that of prednisolone. Known antioxidant, antimicrobial, antiinflammatory, and antispasmodic potentials of Satureja Khuzestanica may be the mechanisms by which this plant protects animals against experimentally induced IBD. Proper clinical investigation should be carried out to confirm the activity in human disease. PMID- 20021010 TI - Effect of adrenalin, adrenochrome, and adrenolutin on connexin proteins in the cardiovasculature. AB - Reported myocardial pathology resulting from increased levels of catecholamines in vivo has led us to investigate the effect of adrenalin on the gap junction proteins connexin 40 (Cx40) and Cx43 and the possible relationship to vascular toxicity. Adrenalin and its known metabolites, adrenochrome and adrenolutin, were used in this study. Utilizing the A7r5 rat aortic cell line, we evaluated the effects of adrenalin, adrenochrome, and adrenolutin on the expression and function of connexin 40 and 43 that are present in both cardiac and vascular tissues. PMID- 20021011 TI - Alterations in L-Histidine Transport in-Response to Aspirin- and Nimesulide Induced Toxicity in Rat Intestine Using Everted Intestinal Sacs. AB - The present study was carried out to investigate the aspirin-and nimesulide induced alteration in the amino acid L-histidine transport across the intestine. These drugs represent the two groups of nonsteroidal anti-inflammatory drugs (NSAIDs) with varying cyclooxygenase-2 (Cox-2) selectivities and are associated with increased risk of gastrointestinal side effects. Female Wistar rats were divided into three different groups: group I (control), group II (aspirin treated, 50 mg/kg), and group III (nimesulide-treated, 10 mg/kg). At the end of 28 days of treatment, the jejunal segment was isolated and everted sacs were prepared so as to study the transport properties of L-histidine and its kinetics. A significant decrease in the transport of histidine in both the treatment groups was seen as compared to the control. Changes in Michaelis-Menten parameters viz: V(max) and the thermodynamic parameter E(a) were also seen. However, K(m) and T(c) remained constant. When studied at different time intervals, both the treatment groups demonstrated an initial rapid transport of L-histidine, which showed saturation at longer durations. Taken together, these results indicate the adverse effects of these two NSAIDs on the intestinal mucosa. Further research, however, is warranted to understand the molecular basis of NSAID-induced GI side effects and to comment decisively on the safety profile of these drugs. PMID- 20021012 TI - A Toxicologist's Guide to the Preclinical Assessment of Hepatic Microsomal Enzyme Induction. AB - The assessment of hepatic microsomal enzyme induction at the completion of preclinical toxicology studies in rodents and large mammals provides a wealth of information to the toxicologist and pharmacokineticist regarding how the drug metabolizing system of the hepatocyte endoplasmic reticulum responded to high dose levels of a xenobiotic designed for a specific pharmacological target in any of several target organs. The interpretation of these data can be greatly enhanced by a clear understanding of how this system functions and what the immediate and long-term ramifications are to organs and organ systems. This review focuses on how drugs modify the hepatic cytochrome P450 system, how those modifications are detected, the various consequences of these modifications, and some differences in the induction response among species. PMID- 20021013 TI - A physiological pharmacokinetic model based on tissue lipid content for simulating inhalation pharmacokinetics of highly lipophilic volatile organic chemicals. AB - The highly lipophilic volatile organic chemicals (HLVOCs) are distributed almost uniquely in the neutral lipid fraction of tissues and blood. As suggested by their high n-octanol:water partition coefficient (>1000), their solubility in water fraction of tissues and blood is negligible. Hypothetically, then, the kinetics of HLVOCs can be simulated solely with the consideration of their solubility and distribution in neutral lipid-equivalent (NLE) fractions of the tissues and blood. The objectives of the present study were therefore (i) to develop a physiological pharmacokinetic model based on NLE content of tissues and blood, and (ii) to apply this model framework for simulating the inhalation pharmacokinetics of HLVOCs (i.e., d-limonene, alpha-pinene, and 1,2,4 trimethylbenzene) in humans. The PBPK model developed in this study consisted of tissue compartments that represented only their NLE content. All biological parameters, except alveolar ventilation rate, were expressed on the basis of their NLE content. Tissue:blood partition coefficients were not used since the solubility of HLVOCs in tissue neutral lipids and blood neutral lipids is considered to be the same. The NLE-based physiological pharmacokinetic model was then used to simulate the uptake and disposition kinetics of alpha-pinene, d limonene, and 1,2,4-trimethylbenzene in humans. The NLE-based model developed in this study represents a novel tool for simulating the lipid concentrations and pharmacokinetics of HLVOCs without the use of tissue:blood partition coefficients. PMID- 20021014 TI - Deaths due to corrosive ingestion: a 10-year retrospective study. AB - OBJECTIVES: Evaluation of the substances used, macroscopic and microscopic autopsy findings, and the manner of corrosive ingestion deaths in Istanbul are aimed in this study. METHODS: The autopsy records of the corrosive ingestion cases in Istanbul, Turkey, between 1992 and 2001 are investigated in our retrospective descriptive study. RESULTS: The cause of death was determined as "corrosive injury" in 23 (0.089%) cases among 25,763 autopsy cases. Among these cases one was in the child age group and the corrosive substance ingestion was accidental, whereas all the other cases were adults and the ingestion was for suicidal intent. Sixteen (69.6%) of the cases were female. Nitric acid was the most frequent agent with 13 cases (56.5%). Necrosis of esophagus was determined in 19 cases (82.6%) and perforation of stomach in 11 cases (47.8%). CONCLUSION: Guidance and education are important tools for prevention from corrosive injuries, but we believe that the best weapon is to restrict easy access to corrosive agents by prohibiting the free commercialization of these most potent agents. PMID- 20021015 TI - Development of functional fish feed with natural ingredients to control heavy metals. AB - The effects of two natural ingredients, Chinese parsley (CP) and chitosan (CT), on growth, accumulation, and excretion of cadmium in fish body and preservation of essential trace metals in the body were investigated by using rainbow trout that had been fed cadmium-added diet, low and high concentration, for 3 weeks. This pretest confirmed that cadmium was accumulated in the liver, kidney, and intestine of the test fish. The cadmium level of the fish, fed diet with CP or CT, was decreased by 18% and 24%, respectively, compared to that of the fish given the control diet. But CP and CT did not have an influence on normal growth of test fish and the levels of essential trace metals in the body. In addition, the level of cadmium was higher in liver than kidney in the high-cadmium dietary group, indicating the Cd level in kidney follows that of liver as kidney lies in the final stage of metabolism. The cadmium accumulation in the fish body was supposed to be reduced, by giving CP to increase the solubility of Cd to body fluid by conjugation into metallothioneins (MTs), while CT was supposed to be responsible for the physical adsorption of cadmium ions by glucosamine groups. PMID- 20021016 TI - Myofibril membranes in relation to the neuromuscular weakness of acute monocrotophos poisoning. AB - Organophosphate poisoning is a common method of deliberate self-harm in countries where the pesticides are readily available. The severity of neuroparalysis and myopathy occurring in acute organophosphate poisoned patients is determined by the severity of poisoning and is associated with morbidity and mortality. Molecular mechanisms that underlie severe paralysis are not well delineated but are essential to know to improve treatment. In this study rats were subjected to increasing doses (0.25 to 0.8 LD(50)) of monocrotophos, and cell membrane lipid profiles, particularly those of myofibril membranes, were examined in relation to neuromuscular weakness occurring in poisoning. Increasing doses of monocrotophos inhibited brain and RBC acetylcholinesterase >/=60% early in the poisoning. RBC acetylcholinesterase levels recovered to 70% to 80% of normal while brain acetylcholinesterase remained 44% to 67% inhibited 1 week after poisoning. Increasing severity of poisoning led to significant changes in myofibril membrane lipid composition with cholesterol to phospholipid ratios increasing from 0.029 +/- 0.008 in controls to 0.063 +/- 0.023 in severe poisoning (p > 0.05). These changes were associated with neuromuscular weakness in the first day of poisoning. Membrane changes were reversible and rats recovered muscle strength in 1 week with no treatment. Lipid compositions of the intestine, brain, and muscle mitochondrial membranes were not affected by monocrotophos. The study indicated that neuromuscular weakness was associated with muscle membrane disorganization early in the course of acute organophosphate poisoning and that subclinical neurotoxicity of long duration may be a consequence of acute organophosphate poisoning. Controlling the severity of poisoning early in the course of acute organophosphate poisoning appears to be important for clinical recovery. PMID- 20021017 TI - Comparison of Elisa- and LC-MS-Based Methodologies for the Exposure Assessment of Bisphenol A. AB - Several types of methods, mainly liquid chromatography (LC), have been used for the analysis and assessment of bisphenol A (BPA) in human biological samples. Enzyme-linked immunosorbent assay (ELISA) is also being used for BPA measurement. In this study, we verified whether ELISA is suitable for measuring human samples, namely, serum and urine, by comparing the ELISA results with those obtained by liquid chromatography with multichannel colometric electrochemical detection (LC/ECD) and liquid chromatography coupled to a mass spectrometer (LC/MS/MS). Results of the measurement with LC/ECD showed urinary BPA concentrations to be 1.92 [1.45] +/- 1.99 (mean [median] +/- standard deviation) ng BPA/mL without the correction of urine volume and 1.20 [0.90] +/- 1.10 mug BPA/g creatinine; however, in serum, free and total BPA were not detected. In both samples, a good correlation of the values with the methods was not found. ELISA is one of the powerful measurement methods, since it is convenient and useful for screening bulk quantities. At this point, however, ELISA is not suitable for BPA measurement in human samples because of low levels of BPA in human samples, matrix effect, and specificity of anti-BPA antibody. PMID- 20021018 TI - In vitro test-based comparison of pesticide-induced sensitivity in marine and freshwater phytoplankton. AB - This study aims to assess the effects of two pesticides, namely the insecticide carbofuran and the herbicide isoproturon, on monospecifically cultivated marine and freshwater phytoplankton according to standard methods. In the presence of pesticide, growth rates were lower in marine species Chaetoceros gracilis and Phaeodactylum tricornutum than in freshwater species Chlorella vulgaris and Selenastrum capricornutum after 72 hours. The EC50 values were obtained with the REGTOX Macro software, and the NEC values by applying the DEBtox model. PMID- 20021019 TI - Safety assessment of a novel niacin-bound chromium-based energy formulation. AB - Over 35 million adults suffer from fatigue or lack of energy. In this study, we assessed the safety of a novel niacin-bound chromium-based Energy Formulation, which also contained caffeine, D-ribose, Withania somnifera extract, and selected amino acids. Niacin-bound chromium is a novel source of bioavailable chromium (III), and known to promote healthy lipid profile. Male and female Sprague-Dawley rats were fed 125 ppm Energy Formulation for 90 consecutive days. Body weight, feed, and water intake were monitored over the period of 90 days. No significant changes were observed between the control and treatment groups following subchronic supplementation with this Energy Formulation. Furthermore, no significant changes were observed in selected organ weights individually and as percentages of body and brain weights. The Energy Formulation supplementation did not cause changes in hepatic lipid peroxidation or DNA fragmentation after 30, 60, or 90 days of treatment. Hematology, clinical chemistry, and histopathological evaluations revealed no adverse effects in the treatment group. These findings demonstrate the safety of this Energy Formulation. PMID- 20021020 TI - Effect of four organophosphorus compounds on human blood acetylcholinesterase: in vitro studies. AB - In the present investigation acetylcholinesterase (AChE) was estimated in erythrocytes from blood samples exposed to four commonly used organophosphorus pesticides in vitro. The aim of the study was to determine the IC50 concentration of the pesticides monocrotophos, chlorpyrifos, profenofos, and acephate as inhibitors of AChE. Cholinesterase was spectrophotometrically measured using acetylthiocholine iodide (AChI) as substrate and dithiobisnitrobenzoic acid (DTNB) as the coloring agent using different aliquots of the pesticides. It was found that as the concentration of the pesticides increased, the inhibition also increased, and a dose-response relationship was obtained in a time-dependent manner. The IC50 values for RBC-AChE were 0.12 muM, 0.25 muM, 0.35 muM, and 4.0 muM for chlorpyrifos, monocrotophos, profenofos, and acephate, respectively. Chlorpyrifos was found to be a more potent inhibitor of AChE followed by the rest of the pesticides used in this study. This investigation determined the relative toxicities of organophosphorus pesticides under laboratory conditions using statistical methods. PMID- 20021021 TI - Hypolipidemic effect of ginger in 1,2-dimethyl hydrazine-induced experimental colon carcinogenesis. AB - Colorectal cancer is increasingly common nowadays in Asian countries and still remains the second leading cause of cancer death in the United States. In our laboratory, we studied the chemopreventive and hypolipidemic effect of ginger, a dietary spice, in 1,2-dimethylhydrazine (DMH)-induced colon cancer. Rats were given a weekly subcutaneous injection of DMH (20 mg/kg body weight), a known colon carcinogen, in the groin for 15 weeks. Ginger (50 mg/kg body weight P.O.) was given at the initiation and also at the postinitiation stages of carcinogenesis. The animals were sacrificed at the end of the experimental period of 30 weeks. The fecal bile acids, neutral sterols, and tissue lipid profile were evaluated using various biochemical estimations. The levels of fecal bile acids, neutral sterols, cholesterol, HMG CoA reductase, free fatty acids, triglycerides, phospholipase A, and phospholipase C were significantly increased, whereas the levels of tissue phospholipids was decreased in DMH-treated rats as compared to control rats. On administering ginger at the initiation and also at the postinitiation stages of colon carcinogenesis, the levels of fecal bile acids, neutral sterols, tissue cholesterol, HMG CoA reductase, free fatty acids, triglycerides, phospholipase A, and phospholipase C were significantly decreased, whereas the levels of phospholipids were increased as compared to unsupplemented DMH treated rats. Thus, ginger supplementation was found to reduce the risk of colon cancer markedly by virtue of its hypolipidemic and antioxidative effects. PMID- 20021022 TI - LHRH-PE40-Induced Vascular Leak Syndrome. AB - AIM: This study was designed to study the toxicity of LHRH-PE40, a recombinant DNA-derived protein composed of the decapeptide known as luteinizing hormone releasing hormone and the translocation and catalytic domains of pseudomonas exotoxin A. METHOD: Single-dose and repeat-dose toxicity of intravenous injection of LHRH-PE40 was studied by clinical signs, hematology, blood chemistry and histopathology, and lung permeability to Evans blue dye. Additional study was performed to find the relationship between dexamethasone pretreatment and vascular leak syndromes. RESULTS: Dyspnea, increased hemocrit, low serum total protein, lung edema, and high lung permeability were found on rats treated with single or repeated doses of LHRH-PE40. Dexamethasone pretreatment before LHRH PE40 administration partly lowered morbidity of rats. CONCLUSION: LHRH-PE40 induced vascular leak syndrome was the chief cause of rats' death. Dexamethasone pretreatment partly reduced the frequency of vascular leak syndrome. Hypotheses about vascular leak syndromes were also formed by reviewing recent literature. PMID- 20021023 TI - Skin penetration by metal compounds with special reference to copper. AB - The process of diffusion into and across the different structures of the skin by chemicals is reviewed, with particular attention given to copper compounds. The scarce data available from the literature indicate that, in contact with the skin, metallic copper will oxidize, and the compounds resulting will penetrate it. Results from our lab confirm that copper compounds formed with skin exudates penetrate the human stratum corneum in a time-dependent fashion. The only quantitative diffusion rates for copper compounds given in the literature so far refer to experiments performed on the cat in vitro and in vivo. Transformation of data from that study, based on certain assumptions, lead to estimated Kp values of 10(-6) to 10(-5) cm/h for the copper salts tested, values that lie at the lower end of skin diffusivity rates measured for transition metal salts. Permeability coefficients for aqueous copper sulfate and acetate through human epidermis in vitro measured in our laboratory are of the order of 10(-6) cm/h. For copper compounds formulated in combination with zinc compounds for therapeutic purposes, applied on dermatomed human skin in vitro in various vehicles, the apparent penetration coefficients Kp were in the range of 3.2 x 10( 6) and 1.6 x 10(-5) cm/h. PMID- 20021024 TI - Changes in Activities of MMP in Alcohol and Thermally Oxidized Sunflower Oil Induced Liver Damage: NAC Antioxidant Therapy. AB - Liver fibrosis is the result of imbalance between extracellular matrix (ECM) synthesis and breakdown. Ethanol-induced increase in redox state is a sign of major change in hepatic metabolism and this inhibits tricarboxylic acid cycle activity and, fatty acid oxidation and increases fatty acid uptake, thus predisposing fatty liver. Fibrotic changes induced by alcohol are provoked by diets rich in PUFA. Heating of oils rich in PUFA produces toxic volatile and nonvolatile compounds, which aggravate liver damage. Hepatotoxicity was induced in male Wistar rats by administering alcohol (20%) and thermally oxidized sunflower oil (Delta PUFA) (15%). When N-acetyl cyteine (NAC) (150 mg/kg body weight), an ROS scavenger, was administered, there was a reversal of liver damage, which was demonstrated biochemically. Matrix metalloproteinases (MMPs), being potential biochemical indicators of fibroproliferation, were estimated in the present study, which were found to be altered in alcohol, Delta PUFA, and alcohol + Delta PUFA. The altered activities of MMPs in these groups were effectively modulated by treatment with NAC. Thus, in this study, NAC was found to modulate the effect of alcohol and Delta PUFA-induced liver damage. PMID- 20021025 TI - Effects of topical phosmet on fur residue and cholinesterase activity of dogs. AB - Fleas, ticks, and mites are a major problem in many areas of the country for pet owners, and one treatment option involves the use of dips that contain pesticides. In the present study, dogs were dipped with a commercial phosmet (Imidan(R)) flea dip using the recommended guidelines for four consecutive treatments to determine the residues available for transfer to humans from the fur of the dogs. Twenty-four dogs of various breeds and weights were dipped, and each animal's fur was sampled with cotton gloves by petting for 5 minutes in a 10'' x 4'' area along the upper back before dipping and at 4 hours, and 1, 3, 7, and 14 days after dipping. Over the 4 dippings the 4-hour samples had a geometric mean of 2653 mug, and the 1-, 3-, 7-, and 14- day samples had geometric means of 877, 316, 84, and 20 mug, respectively. The samples ranged (in mug) from 80 to 16,794 at 4 hours, 44 to 7028 at 1 day, 1 to 4897 at 3 days, 1 to 2691 at 7 days, and 0.3 to 835 at 14 days. The residues removed by the petting did increase with the subsequent dips, but this was probably due to handler experience. The increase is not attributed to accumulation since there was less than 2% of transferable residue on the dog at 14 days post application. There was no significant inhibition of the plasma cholinesterase in the dogs over the study, suggesting that there was either a very low level of dermal absorption of phosmet or there was rapid detoxication (supported by EPA R 825170-01-0). PMID- 20021026 TI - Does 5-hydroxytryptophan cause acute and chronic toxic perturbations in rats? AB - In 1989, an epidemic of eosinophilia-myalgia syndrome (EMS) occurred in the United States that was attributed to contaminated l-tryptophan (LT). Features of tryptophan-induced EMS included debilitating myalgia and marked peripheral eosinophilia. Although the contaminant(s) was found only in the product produced by a single manufacturer (Showa Denko), all LT was withdrawn from the market and replaced by 5 hydroxytryptophan (5HTP). The belief was that the latter should not contain the implicated contaminant(s), because it was manufactured by a process entirely different from the banished LT. Nevertheless, in 1994 a case diagnosed as EMS appeared. Although the exact causative factor(s) in LT and the possible 5 HTP-induced EMS are uncertain, many reported finding "Peak E" in contaminated LT and the presence of "Peak X" in the 5-HTP of the 1994 case. The latter finding led some to assume that Peak X was a potential pathological agent in 5-HTP that might cause future cases of EMS. To determine whether 5-HTP could cause EMS, we followed 120 male Sprague-Dawley rats, 7 to 8 weeks of age (body weight 200-250 g), for 1 year. They were divided into three groups of 40. One group acted as control, drinking only water; a second group received a low dose of 5-HTP in their drinking water (87.5 mg/dL); and the last group drank a high dose of 5-HTP, 875 mg/dL. No significant differences in the body weights of these three groups of animals were observed over the year. After 2 months, systolic blood pressures (SBP) in the 5-HTP groups were significantly lower for the duration of the study. At the end of 12 months, SBP of the control group averaged 140 mm Hg, the low dose 5-HTP group averaged 133 mm Hg, and the high-dose group averaged 125 mm Hg. Even though enough 5-HTP was given to cause a physiological response, no significant differences were found in the hematological values, including eosinophil count. Also, no significant differences were found in hepatic and renal values. In the histological studies, no treatment-related changes were noted in the hearts, livers, pancreases, leg striated muscles, and small intestines. In particular, there was no evidence of eosinophilic infiltration and fascial/perimysial inflammation. Accordingly, no significant evidence of EMS was seen in rats receiving high-dose 5-HTP for 1 year. PMID- 20021027 TI - Development of a recombinant yeast assay to detect ah-receptor ligands. AB - Endocrine systems of humans and animals are disturbed by dioxin-like compounds, which are ligands of the aryl hydrocarbon receptor (AhR). It is important to determine the accumulation of dioxin-like compounds in the environment for maintenance of human health. In this study, we developed a new method for screening ligands of the AhR using a yeast hybrid system. Reporter genes constructed by the insertion of dioxin response elements were integrated into HIS and lacZ yeast genomes. Then yeast was transformed with GAL4-activated domain fused AhR and aryl hydrocarbon receptor nuclear translocator expression constructs. At 10(-4) M of beta-naphthoflavone, which is an AhR ligand, the absorbance of optical density at 600 nm (OD 600) and beta-galactosidase activity was significantly increased. beta-galactosidase activity was increased when the concentration of 3-methylcholanthrene (MC) was increased. ATP concentration increased as concentration of MC increased up to 10(-10) M but decreased at higher concentrations. The concentration of ATP in the cell suspensions increased linearly with OD 600, used as an index of cell density (r(2) = 0.8366, F = 209.9, p < 0.0001, n = 44). The established yeast assay could possibly be used in the future to detect dioxin-like compounds in environmental samples. PMID- 20021028 TI - Development of a microinstillation model of inhalation exposure to assess lung injury following exposure to toxic chemicals and nerve agents in Guinea pigs. AB - Respiratory disturbances due to chemical warfare nerve agents (CWNAs) are the starting point of mass casualty and the primary cause of death by these weapons of terror and mass destruction. However, very few studies have been implemented to assess respiratory toxicity and exacerbation induced by CWNAs, especially methylphosphonothioic acid S-(2-(bis(1-methylethyl)amino)ethyl)O-ethyl ester (VX). In this study, we developed a microinstillation technique of inhalation exposure to assess lung injury following exposure to CWNAs and toxic chemicals. Guinea pigs were gently intubated by placing a microcatheter into the trachea 1.5 to 2.0 cm centrally above the bifurcation. This location is crucial to deliver aerosolized agents uniformly to the lung's lobes. The placement of the tube is calculated by measuring the distance from the upper front teeth to the tracheal bifurcation, which is typically 8.5 cm for guinea pigs of equivalent size and a weight range of 250 g to 300 g. The catheter is capable of withstanding 100 psi pressure; the terminus has five peripheral holes to pump air that aerosolizes the nerve agent that is delivered in the central hole. The microcatheter is regulated by a central control system to deliver the aerosolized agent in a volume lower than the tidal volume of the guinea pigs. The average particle size of the nerve agent delivered was 1.48 +/- 0.07 micrometer. The microinstillation technology has been validated by exposing the animals to Coomassie brilliant blue, which showed a uniform distribution of the dye in different lung lobes. In addition, the concentration of the dye in the lungs correlated with the dose/time of exposure. Furthermore, histopathological analysis confirmed the absence of barotraumas following micoinstillation. This novel technique delivers the agent safely, requires less amount of agent, avoids exposure to skin, pelt, and eye, and circumvents the concern of deposition of the particles in the nasal and palette due to the switching of breathing from nasal to oronasal in whole-body dynamic chamber or nose only exposure. Currently, we are using this inhalation exposure technique to investigate lung injuries and respiratory disturbances following direct exposure to VX. PMID- 20021029 TI - Alterations in oxidative stress-related parameters in rat testis following monensin administration. AB - Monensin, a carboxylic ionophore, is well known for Na(+)/H(+) exchanger activity across biological membranes. It is also used in the poultry industry for its useful effects as a food additive. The present study has been designed to investigate the effects of monensin on some oxidative stress-related parameters in rat testis. Monensin was administered intratesticularly (5 mug/testis) to both testes by a single dose to Wistar rats for different time periods. After the completion of the respective treatments, various parameters reflecting the antioxidant defense system of the tissue were monitored and marked changes were found in the activities of various enzymes as well as in the levels of reduced glutathione and lipid peroxidation. After 1, 2, 3, and 4 days of monensin treatment, the activity of superoxide dismutase was found to be unaltered. However, after 2 days of monensin treatment, glutathione-S-transferase and catalase showed inhibition in their activities along with the depletion of glutathione (reduced) accompanied by a marked increase in lipid peroxidation. The increase in lipid peroxidation was noticeable even after 1 day of monensin administration. The inhibition in glutathione-S-transferase and glutathione peroxidase activities was also observed along with an increase in lipid peroxidation at the end of the 3-day posttreatment period, while, the 4-day posttreatment schedule caused an increase in the activity of glutathione reductase and glutathione peroxidase that was also accompanied by an inhibition of catalase. The findings of the present study are indicative of the potential of monensin in testicular tissue in contraceptive intervention. PMID- 20021030 TI - Optimization of the (32)P-Postlabeling/Thin Layer Chromatography Assay ((32)P TLC) for In Vitro Detection of 8-Oxo-Deoxyguanosine as a Biomarker of Oxidative DNA Damage. AB - During the last years, much attention was focused on the measurement of 8-oxo-7,8 dihydro-2'-deoxyguanosine (8-oxo-dG) as a marker of oxidative DNA damage. Among various analytical techniques, the (32)P-postlabeling assay has been applied in determination of 8-oxo-dG. However, artefactual DNA oxidation could take place during the work-up procedures leading to over-estimate the level of 8-oxo-dG. In the present study, we optimized the (32)P-postlabelling assay with thin layer chromatography to measure 8-oxo-dG in standard samples of 8-oxo-dG, calf thymus DNA and primary cultured rat hepatocytes. The background levels of 8-oxo-dG in calf thymus DNA and in primary cultured rat hepatocytes were lesser than those determined by the previously described (32)P-postlabeling procedures and were in the range of those determined by chromatography methods (GC-MS, HPLC-MS). PMID- 20021031 TI - Mitochondrial Bioenergetics, Diabetes, and Aging: Top-Down Analysis Using the Diabetic Goto-Kakizaki (GK) Rat as a Model. AB - In the present study we investigated the changes in the oxidative phosphorylation system of liver mitochondria, isolated from diabetic Goto-Kakizaki (GK) and Wistar (control) rats with different ages (6, 12, 26, and 52 weeks). We used a kinetic approach known as "top-down" analysis, which conceptually divides the oxidative phosphorylation system into two subsystems: one producing the protonmotive force (Deltap) and another that consumes Deltap. The overall response of the Deltap generators to Deltap was obtained from an uncoupler titration of respiration rate versus Deltap, while the overall response of Deltap consumers to Deltap was obtained from an inhibitor titration of respiration rate versus Deltap. Our results showed that GK liver mitochondrial preparations presented an increase in Deltap production and phosphorylative subsystems (using succinate as respiratory substrate). The alterations observed may suggest the existence of biochemical compensatory mechanisms to type 2 diabetes mellitus in GK rats during their first year of life, in order to reduce the injury associated with the disease. Furthermore, we observed that liver metabolic efficiency of mitochondrial respiration declined with age, this decrease in respiratory activity being visible both in control and diabetic rats. PMID- 20021032 TI - Prevention by L-arginine/nitric oxide of chlordiazepoxide-induced toxic reactions in the rat salivary gland. AB - The effects of chlordiazepoxide, L-Arginine (nitric oxide precursor), L-NAME (nitric oxide synthesis inhibitor), and concurrent treatment of these drugs on concentration of salivary total protein, calcium, sodium, potassium concentrations, amylase activity, and flow rate in rats were studied. Total saliva was collected 2 hours postintraperitoneal administration of chlordiazepoxide (5 mg/kg), L-Arginine (250 mg/kg), and L-NAME (10 mg/kg) intraorally by polyethylene cannula from anaesthetized rats for 30 min using pilocarpine (6 mg/kg) as secretagogue. Chlordiazepoxide reduced flow rate to 43%, total protein concentration to 30%, calcium concentration to 33%, and amylase activity to 33% of control. Potassium and sodium levels were not affected by chlordiazepoxide. L-Arginine increased flow rate to 10%, total protein to 65%, and calcium to 17%, while L-NAME decreased flow rate to 9%, total protein to 55%, and calcium to 25%. Concurrent treatment of rats by chlordiazepoxide plus L Arginine or L-NAME influenced chlordiazepoxide-induced alterations in saliva composition. The inhibitory effects of chlordiazepoxide on salivary flow rate, amylase activity, and concentrations of total protein and calcium were blocked by L-Arginine while potentiated by L-NAME coadministrations. It is concluded that benzodiazepines induce toxicity in salivary gland function by disturbing the homeostasis of inositol triphosphate and Ca homeostasis in the cells. Administration of L-Arginine prevents benzodiazepine-induced toxicity most probably through its vasoregulatory action and balancing intracellular Ca homeostasis. PMID- 20021033 TI - Spectrophotometric Analysis of the Cyanide Metabolite 2-Aminothiazoline-4 Carboxylic Acid (ATCA). AB - Methods of directly evaluating cyanide levels are limited by the volatility of cyanide and by the difficulty of establishing steady-state cyanide levels with time. We investigated the measurement of a stable, toxic metabolite, 2 aminothiazoline-4-carboxylic acid (ATCA), in an attempt to circumvent the challenge of directly determining cyanide concentrations in aqueous media. This study was focused on the spectrophotometric ATCA determination in the presence of cyanide, thiocyanate (SCN(-)), cysteine, rhodanese, thiosulfate, and other sulfur donors. The method involves a thiazolidine ring opening in the presence of p (hydroxy-mercuri)-benzoate, followed by the reaction with diphenylthiocarbazone (dithizone). The product is spectrophotometrically analyzed at 625 nm in carbon tetrachloride. The calibration curve was linear with a regression line of Y = 0.0022x (R(2) = 0.9971). Interference of cyanide antidotes with the method was determined. Cyanide, thiosulfate, butanethiosulfonate (BTS), and rhodanese did not appreciably interfere with the analysis, but SCN(-) and cysteine significantly shifted the standard curve. This sensitive spectrophotometric method has shown promise as a substitute for the measurement of the less stable cyanide. PMID- 20021034 TI - Proteomic analysis to identify the cellular responses induced by hydroquinone in human embryonic lung fibroblasts. AB - Hydroquinone (HQ), a major metabolite of benzene, is used widely as a reagent in photographic developers, as an antioxidant in the manufacture of rubber, as a polymerization inhibitor for acrylic and vinyl acetate monomers, and in cosmetic products as a skin-lightening agent. But the mechanism of its effect on human cells is far from clear. In the present work, we studied the cellular response induced by HQ using proteomic approaches. Human embryonic lung fibroblasts (HLFs) were treated with 100 mu M HQ for 24 h. This dose of HQ was found in assays to significantly decrease cell viability. After treatment, two-dimensional electrophoresis was performed using the Amersham Bioscience 2DE system following the manufacturer's instructions. Proteins were visualized by staining with colloidal coomassie blue. Fifteen protein spots showed significant changes after HQ treatment. Eleven protein spots were identified by peptide mass fingerprinting using MALDI-TOF or by peptide sequence tagging using MALDI-TOF-TOF. Among them are transaldolase, growth factor receptor-bound protein 2, mutant beta -actin, gamma -actin, Lasp-1, TAR DNA-binding protein, and a protein similar to neural precursor cell-expressed protein. These include proteins involved in oxidative stress, cellular signaling, RNA splicing, and cytoskeleton reconstruction. Most of their involvements in the cellular responses to HQ have not been reported. Therefore, our findings may offer new insights into the mechanisms of HQ cytotoxicity and these proteins may serve as new biomarkers for detecting exposure of human populations to HQ. It is suggested that proteomic approaches may provide new strategies to evaluate the toxicity of xenobiotics. PMID- 20021035 TI - Normal Development of Reproductive System in Rat Male Offspring Following Perinatal Combined Exposure to p,p'-DDE and 1,4-Dichlorobenzene. AB - Prenatal or early postnatal exposure to some synthetic chemicals may affect the later reproductive system of the offspring, and there may also be unique responses observed due to exposure to combinations of chemicals that are not observed when the chemicals are present individually. Organochlorine compounds are ubiquitously distributed in the environment. p,p'-DDE (1,1-dichloro-2,2 bis (p-chlorophenyl) ethylene) is a persistent metabolite of DDT and 1,4 dichlorobenzene (DCB) is used as an air freshener or a moth repellent. Oral p,p' DDE intake through marine products and DCB concentrations of residential air are demonstrated to be high in Japan. Consequently, the fetus and neonate are supposed to be exposed much more to p,p'-DDE and DCB via the maternal body. Therefore, effects of perinatal combined exposure to p,p'-DDE and DCB on the male reproductive system after maturation have been investigated in rat male offspring of dams ingesting these contaminants during the perinatal period from gestational day 1 to postpartum day 21. In this study, no deteriorated developmental effects have been observed in rat male offspring until maturation following oral administration of 125 ppm p,p'-DDE (approximately 10 mg/kg) and 25 ppm DCB (approximately 2 mg/kg) to dams. Further, no obvious effects of perinatal combined exposure to DCB and p,p'-DDE on the male reproductive organs and sperm parameters were observed in mature male offspring. PMID- 20021036 TI - Energy and redox States in the c6 glioma cells following acute exposure to zn, se(+4), and se(+6) and the correlation with apoptosis. AB - Changes in cellular energy and redox states in the C6 glioma cells exposed to increasing concentrations of either Zn or Se were studied to examine whether different elements cause different patterns of changes in cellular metabolism. Following a 3-h exposure, both Zn and Se(+4) caused dose-dependent decreases in cell viability and total adenosine nucleotides (TAN = ATP + ADP + AMP). In addition, Zn caused a dose-dependent increase in cellular ATP/TAN and a decrease in the ADP/TAN and AMP/TAN. These changes resulted in a significant increase in energy charge potential (ECP = [ATP + 0.5ADP]/TAN). Se(+4), on the other hand, caused a dose-dependent decrease in ATP/TAN but an increase in both ADP/TAN and AMP/TAN, resulting in a dose-dependent decrease in ECP. Both Zn and Se(+4) caused a dose-dependent decrease in GSH/GSSG and an increase in GSH + GSSG when compared to TAN. In contrast to Zn and Se(+4), the nontoxic Se(+6) caused no significant changes in cellular energy states but reduced the GSH/GSSG ratio from 3.14 +/- 0.49 to 2.05 +/- 0.29, which could be explained by the effect of Se on enzymes responsible for GSH metabolism. As the cellular ATP level has been considered an important element that mediates the mode of cell death, it was suggested that a significant increase in ATP/TAN upon exposure to Zn would indicate that cell death occurred via apoptosis, while Se(+4) caused a different pattern of cell death. This was confirmed by the appearance of cells with fragmented nucleus in cells treated with Zn, but not Se(+4) and Se(+6). The results demonstrated that different chemicals caused different patterns of metabolic changes. The correlation between metabolic changes and the mode of cell death was discussed. PMID- 20021037 TI - Aluminium-induced imbalance in oxidant and antioxidant determinants in brain regions of female rats: protection by centrophenoxine. AB - The present study was carried out to investigate the potential of centrophenoxine in modulating aluminium-induced neurotoxicity. Female Sprague Dawley rats were administered aluminium chloride orally (40 mg/kg b.w./day) for a period of 8 weeks. At the end of respective treatment, various markers of oxidative stress were determined in four different regions of brain: cerebrum cerebellum, medulla oblongata, and hypothalamus. Lipid peroxidation assay was also carried out using standard techniques. Simultaneously, the centrophenoxine group (100 mg/kg b.w./day) for 6 weeks was also run long to understand the role in ameliorating oxidative damage. A significant decrease in the activities of superoxide dismutase and catalase was noticed in all the four regions, the most significant being in the hypothalamus (0.603 +/- .06) and cerebrum (0.038 +/- .01). Due to aluminium toxicity, peroxidation of lipids was also found to be elevated in cerebrum (0.424 +/- .03), cerebellum (0.341 +/- .03), hypothalamus (1.018 +/- .007), and medulla oblongata (0.304 +/- .05). However, posttreatment with centrophenoxine significantly elevated the superoxide and catalase activities in different regions. In addition, lipid peroxidation status of membranes was significantly reduced after centrophenoxine posttreatment to aluminium-exposed animals. Centrophenoxine has proved to be beneficial in combating the damage caused by aluminium toxicity. However, further research is needed to have a better understanding of the molecular basis of aluminium-induced oxidative damage. In addition, the different aspects of centrophenoxine need to be unmasked. PMID- 20021038 TI - A General Physiological and Toxicokinetic (GPAT) Model for Simulating Complex Toluene Exposure Scenarios in Humans. AB - Realistic simulation of environmental exposure scenarios requires dynamic methods in which exposures and human activities vary continuously as a function of time. Simulation of such complex scenarios is, with conventional physiologically based methods, a complex and programming-intensive task. The goal of the present effort was to simplify this task by combining a commercially available general whole body human physiological model (QCP2004) with a slightly extended physiologically based toxicokinetic (PBTK) model from the literature. The QCP2004 model is a differential equation-based model similar to PBTK models except that normal organ function is simulated and the body organs are appropriately interlinked. Here QCP2004 provided estimates of physiological parameters required by the PBTK model. These were updated as the model was iteratively executed appropriate to the varying activity of the human subject. The combined general physiological model and the PBTK model was called a general physiological and toxicokinetic (GPAT) model. The GPAT model was tested and (within the constraints of available toluene exposure experiments in the literature) found to predict toluene blood concentrations, even in dynamic situations. A model of the structure used in the present work is capable of expansion as new knowledge is developed and greater detail is desired. Similarly, multiple toxicant PBTK models can be developed and incorporated for applications to mixtures risk assessment. Additionally, toxicant effects on organ systems can be achieved by altering organ function during a simulation as a function of the internal dose of toxicants. By cumulatively adding detail to the model as new physiological and chemical-specific information becomes available, the model can become a repository of knowledge for increasingly sophisticated risk-assessment applications. PMID- 20021039 TI - Effectiveness of Zinc in Modulating the CCl (4) - Induced Oxidative Stress in Rat Liver. AB - The present study was designed to evaluate the role of zinc on the enzymes involved in oxidative defense mechanism in conditions of carbon tetrachloride (CCl (4)) -induced rat liver toxicity. To carry out the study, rats were divided into four groups: group I (normal control), group II (CCl (4) treated), group III (zinc control), and group IV (zinc + CCl (4) treated). Animals in group II and group IV were administered 0.1 mL of CCl (4) mixed with 0.1 mL of groundnut oil on alternate days for a period of 8 weeks. Zinc in the form of zinc sulphate was given to animals of group II and group IV at a dose level of 227 mg/L in drinking water. Animals given CCl (4) treatment alone showed a significant increase in the activities of superoxide dismutase (SOD), catalase, glutathione peroxidase (GPx), and the concentration of malondialdehyde (MDA) product, whereas a significant depression was observed in the levels of reduced glutathione (GSH). Zinc treatment to CCl (4) -treated rats brought the altered levels of GSH, MDA, and SOD as observed following CCl (4) -treatment alone to within normal limits. However, the activities of catalase and glutathione peroxidase, which were increased under carbon tetrachloride treatment, were significantly attenuated and tended to become normal. Hence, the present study suggests that zinc may play an important role in regulating the activities of the enzymes involved in antioxidative defense system under CCl (4) toxic conditions. PMID- 20021040 TI - Delta-aminolevulinic Acid dehydratase genotype and its relationship with blood lead and zinc protoporphyrin levels in lead-exposed children living in a smelter community in northern Mexico. AB - The implications of delta-aminolevulinic acid dehydratase (ALAD) polymorphism for lead kinetics and toxicity have been mainly studied in occupationally exposed adults. Therefore, our purpose was to evaluate the distribution of ALAD genotype and its association with biomarkers of exposure (PbB levels) and effect (Blood ZPP) among children living in a smelter community in Mexico. We recruited 569 children from nine elementary schools close to a smelter site. PbB was determined by electrothermal atomic absorption spectrometry. A polymerase chain reaction (PCR)-based protocol was used for ALAD genotyping. Zinc protoporphyrin (ZPP) in blood was measured by direct fluorometry. Most children (93.15%) were homozygous for ALAD (1-1), 6.67% were heterozygous for ALAD for (1-2), and one child was homozygous for ALAD (2-2). There was an increased proportion of ALAD (1-2/2-2) genotype with respect to PbB levels. The ZPP geometric mean was slightly higher in ALAD (1-1) genotype children (63.48 mu mol ZPP/mol Hb) than in those having the ALAD-2 genotype (58.22 mu mol ZPP/mol Hb; p = 0.051). Linear and quadratic models showed significant relationships between ZPP and PbB. A significant increase in the odds ratio (OR) for the effect of lead exposure on ZPP levels was observed for ALAD (1-1) children having PbB values above 20 mu g/dL, as compared to those having PbB levels below 10 mu g/dL (OR = 2.95, 95% CI = 1.45-5.97; p = 0.003), whereas no significant increases were observed for the ALAD (1-2/2-2) children. In summary, our results suggest that heme biosynthesis was less affected in ALAD (1-2/2-2) lead-exposed children than in those carrying the ALAD (1-1) genotype. PMID- 20021041 TI - Stimulatory effects of malathion on the key enzymes activities of insulin secretion in langerhans islets, glutamate dehydrogenase and glucokinase. AB - Previous studies showed that malathion induces hyperglycemia mainly through influence on glucose metabolism in liver and skeletal muscles. The main objective of the present study was to determine what will happen on pancreatic key enzymes of insulin secretion, including glucokinase (GK) and glutamate dehydrogenase (GDH), if animals would be in acute or subchronic exposure to various doses of malathion, an organophosphorous insecticide in rats. In the subchronic study, malathion was administered orally at doses of 100 to 400 ppm for 4 weeks. In the acute experiment, animals received various doses of 3 to 75 mg/kg of malathion intraperitoneally. In each experiment, islets were isolated from the pancreas of rats by a standard collagenase digestion, separation by centrifugation, and hand picking technique. The activities of the mitochondrial GDH and the nonmitochondrial GK enzymes were determined in islets homogenates spectrophotometrically. Blood sample was taken by cardiac puncture for glucose and insulin assays. In the acute experiment, malathion (3, 15, 75 mg/kg) increased blood glucose and insulin (15 and 75 mg/kg). In the subchronic experiment, malathion (100, 200, 400 ppm) increased blood glucose and insulin (200 and 400 ppm). All doses in both acute and subchronic experiments increased the mitochondrial GDH activity. Acute (15 and 75 ppm) and subchronic (200 and 400 ppm) increased the islets GK activity. It was concluded that pancreatic islet key enzymes are stimulated following acute and subchronic exposure to malathion though not enough to overcome to hyperglycemia. Activation of islets muscarinic receptors by malathion in favor of hyperinsulinemia, overproduction of glutamate/glutathione by GDH, and overproduction of glucose via increased glycogenolysis in counteracting with malathion-induced oxidative stress are possible mechanisms for observed effects. A new NOAEL acceptable daily intake must be established for malathion. PMID- 20021042 TI - Rhodanese in mouse brain: regional differences and their metabolic implications. AB - Our results concern the regional localization of rhodanese in the mouse brain. A histoenzymatic examination was undertaken in telencephalon, diencephalon, mesencephalon, and rhombencephalon. The sites of rhodanese activity are revealed as punctuate, granular, dark dots, small in some regions such as hippocampus or bigger in others, and as long, thread-like particles especially, abundant in the region of the telencephalon in the astroglia cells and in the region of the mesencephalon in the hippocampus. There were sites with a high density of the histochemical test products, for example, the ependymoma of the forth cerebral ventricle, choroid plexus, and nerve ducts. These findings support the detoxifying role of rhodanese in brain regions. PMID- 20021043 TI - Nicotinamide prevents apoptosis in human cortical neuronal cells. AB - In previous studies, the free radical generating toxin tertiary butylhydroperoxide (t-BuOOH) was found to induce significant cell death in human cortical neuronal cells (HCN2 cells). Pretreatment with the poly (ADP-ribose) polymerase (PARP) inhibitor nicotinamide was able to prevent HCN2 cell death. In this study it is observed that apoptosis is induced following the addition of t BuOOH at 6 h as indicated by TUNEL-positive cells. When nicotinamide is added prior to t-BuOOH, it is able to prevent neuronal cell death and inhibit apoptosis. DNA microarray studies demonstrate that t-BuOOH administration causes an upregulation of proapoptotic genes like ICH-2 and BimL. On the other hand, nicotinamide-pretreated neurons have higher expression levels of inhibitors of apoptosis (IAP) genes. Therefore, it appears that one mechanism by which nicotinamide acts as neuroprotective agents is by elevating the gene expression levels of IAPs. Moreover, there is an upregulation of the glyceraldehydes-3 phosphate dehydrogenase gene in nicotinamide-pretreated HCN2 cells. Nicotinamide pretreated cells also had higher expression levels of putative "death domain" genes like p75TNFR, TRAIL2, TNFR1, and HVEM-L. Thus, nicotinamide can regulate multiple apoptotic genes with seemingly opposite roles and through its action on these various genes prevent apoptosis of neuronal cells. PMID- 20021044 TI - Amifostine-antioxidant drug in anticancer therapy. AB - Antineoplastic drugs and/or their metabolites are very reactive chemically and easily react with cell components, both those neoplastic and healthy, which leads to uncontrolled cell damage. Research is being carried out to discover new drugs preventing those destructive changes in healthy cells. Amifostine is one of the few drugs with protective abilities in regard to healthy tissues during antineoplastic therapy. Amifostine alone is inactive chemically, but active metabolite amifostine WR-1065 has been assumed to protect the healthy tissues during antineoplastic therapy by bounding of anticancer drug, causing their detoxification and/or eliminating the free radicals generated during radiation and cytostatic therapy. WR-1065 also protects the healthy tissues by inhibition of apoptosis caused by irradiation and/or alkylating compounds. However, the protective mechanism of amifostine influence on healthy cells has not been fully comprehended, and the research is still being carried out. PMID- 20021045 TI - The Tg.AC Transgenic Mouse as a Screening Tool for Anticarcinogens: Broccoli Juice Protected Against 12-O-Tetradecanoylphorbol-13-Acetate (TPA) But Not Benzo[a]Pyrene (B[a]P)-Induced Skin Tumors. AB - The Tg.AC mouse model was used to assess the utility of a short-term screening assay to evaluate compounds with suspected carcinogenic/anticarcinogenic activity. Crude broccoli juice (BROC) was evaluated for antitumorigenic effects against 12-Otetradecanoylphorbol-13-acetate (TPA) and benzo[a]pyrene (B[a]P) induced tumors. Groups of female mice were dosed three times a week (200 muL/mouse) with one of the following: acetone vehicle control, TPA (2.5 mug/mouse), B[a]P (250 mug/mouse), BROC/TPA, or BROC/B[a]P. BROC (200 muL, 1:1 acetone) was dermally administered 1 h prior to the administration of either TPA or B[a]P to evaluate anticarcinogenic activity. Papilloma numbers were recorded weekly for each mouse. Following 13 weeks of treatment, samples from the dermal test site from all mice were examined histologically. B[a]P-induced tumors were evaluated for transgene expression by RT-PCR and immunohistochemically for cyclin D1 and p53 proteins. TPA and B[a]P induced tumors in all surviving mice. BROC showed effective antitumorigenic activity against TPA but not B[a]P. Tumor development was distinct between TPA (small, benign papillomas) and B[a]P (large, ulcerated, squamous cell carcinomas). The transgene v-Ha-ras, cyclin D1, and p53 proteins were highly expressed in B[a]P tumors where progression to malignancy was rapid (< 13 weeks). The effects induced by B[a]P appeared to cooperate with transgene expression to enhance conversion to malignancy and could serve as a phenotypic indicator for genotoxic versus nongenotoxic carcinogens. The model distinguished differences in tumor response for carcinogenic and anticarcinogenic agents. The Tg.AC mouse model offers a potentially useful screen for identifying new anticarcinogenic agents and directing future mechanistic evaluations. PMID- 20021046 TI - Safety and Toxicological Evaluation of a Novel, Standardized 3-O-Acetyl-11-keto beta-Boswellic Acid (AKBA)-Enriched Boswellia serrata Extract (5-Loxin(R)). AB - The novel anti-inflammatory properties of the gum resin derived from Boswellia serrata, also known as Salai guggal in Ayurvedic medicine, are well recognized and highly recommended for human consumption. The active constituents of the gum resin are boswellic acids (BAs). Among the BAs, AKBA potently inhibits 5 lipoxygenase product formation with an IC(50) of 1.5 m muM. We developed a novel Boswellia serrata extract (5-Loxin(R)) enriched with 30% AKBA (US Patent 2004/0073060A1). The genetic basis of the anti-inflammatory effects of 5-Loxin(R) was explored in a system of TNFalpha-induced gene expression in human microvascular endothelial cells. 5-Loxin(R) significantly prevented the TNFalpha induced expression of matrix metalloproteinases and adhesion molecules (ICAM-1 and VCAM-1), and inducible expression of the mediators of apoptosis. With such interesting findings, we planned to determine the broad-spectrum safety of 5 Loxin(R). Acute oral, acute dermal, primary skin and eye irritation, and dose dependent 90-day subchronic toxicity studies were conducted. In safety studies, acute oral LD(50) of 5-Loxin(R) was found to be greater than 5,000 mg/kg in both male and female Sprague-Dawley rats. No changes in body weight or adverse effects were observed following necropsy. Acute dermal LD(50) of 5-Loxin(R) was found to be >2,000 mg/kg. Primary skin irritation test was conducted with 5-Loxin(R) on New Zealand Albino rabbits and 5-Loxin(R) was classified as nonirritating. Primary eye irritation test was conducted with 5-Loxin on rabbits and 5-Loxin(R) was classified as mildly irritating to the eye. A dose-dependent 90-day subchronic toxicity study demonstrated no significant changes in selected organ weights individually and as percentages of body and brain weights. 5-Loxin(R) supplementation did not cause changes in hepatic DNA fragmentation on 30, 60, or 90 days of treatment. Hematology, clinical chemistry, and histopathological evaluations did not show any adverse effects in all organs tested. Taken together, these results demonstrate the broad spectrum safety of 5-Loxin(R). PMID- 20021047 TI - The water disinfectant byproduct dibromoacetonitrile induces apoptosis in rat intestinal epithelial cells: possible role of redox imbalance. AB - Chemically induced oxidative stress poses cytotoxic effects on intestinal epithelial cells that may trigger various forms of injuries in intestinal mucosa. Haloacetonitriles, including dibromoacetonitrile (DBAN, a drinking water contaminant and direct acting mutagen and carcinogen), are known to induce GI disorders. Earlier, we showed that dichloroacetonitrile (DBAN analog) alters glutathione status and increases reactive oxygen species in murine macrophage cell line. Therefore, the present study was undertaken to understand the role of redox imbalance and apoptosis in DBAN-induced GI disorders using rat intestinal epithelial (RIE) cells. Cultured confluent monolayers of RIE cells were continuously exposed to DBAN at 50 to 400 ppb (0.6 to 4.4 muM). After 24, 48, and 72 h of the exposure, oxidative stress and apoptosis were determined. At higher exposure regimens (100 to 400 ppb), a concentration- and time-dependent increase in glutathione disulfide levels (1.5 to 4-fold and 1.6 to 5-fold, p < 0.05) was noticed at 48 and 72 h, respectively, as compared to control. Severe depletion of reduced glutathione was also observed at 72 h after DBAN treatment. DBAN-induced oxidative stress was demonstrated at all concentrations by increased malondialdehyde (MDA) levels (1.3 to 3- and 1.8 to 4-fold, p < 0.05) at 48 and 72 h after treatment, respectively. Increase (1.3 to 2-fold, p < 0.05) in 8-hydroxy 2-deoxyguanosine (8OHdG) levels was observed at 48 h after treatment with 100-400 ppb DBAN. At 72 h these levels were 1.7 to 3- fold higher in DBAN-treated RIE cells as compared to control. DBAN-induced apoptosis, evaluated using TUNEL assay and differential staining techniques, indicates an increase in nuclear damage along with various apoptotic features using epifluorescence or light microscopy. The results of the present study suggest that DBAN-induced redox imbalance could lead to apoptosis and overall oxidative stress in RIE cells. PMID- 20021049 TI - Errata. PMID- 20021048 TI - Effects of Piperonyl Butoxide on the Metabolism of DEF S,S,S-Tributyl Phosphorotrithioate) in Fingerling Channel Catfish. AB - The present study was undertaken to investigate the significance of monooxygenases in bioactivation of DEF to a more effective anticholinesterase in fish. Channel catfish were exposed via the water column for 20 h to piperonyl butoxide (PBO) followed by a 4-h exposure to the organophosphate defolient DEF (concurrent with the PBO). Acetylcholinesterase (AChE) and aliesterases (ALiEs) activities were determined at 0 and 12 h after the exposure period. Inhibition of brain, liver, and plasma AChE activity by DEF was antagonized by PBO; muscle AChE was not inhibited by DEF. Piperonyl butoxide did not antagonize the inhibition of liver or plasma ALiEs by DEF. These results suggest that PBO retards the formation of the metabolite(s) of DEF that inhibit AChE, and that DEF is an effective inhibitor of ALiEs without metabolic activation. PMID- 20021050 TI - Errata. PMID- 20021051 TI - Effects of dengue virus infection on the spleen of male mice given hexavalent chromium with drinking water. AB - The present study was undertaken to investigate the effects of dengue virus (DV) infection in male mice given drinking water containing 250 ppm Cr (VI) and the normal control male mice given plain water to drink. On the basis of intake of water in 24 h, the average dose of Cr (VI) in each mouse was 14.8 mg/kg. After 3, 6, and 9 weeks of drinking Cr (VI), a set of five mice from each group were inoculated intracerebrally (ic) with a 1000 x LD(50) (100 times the lethal dose that kills 50% mice) dose of DV, and the effects on the spleen were studied at the fourth and eightth day postinoculation. It was observed that Cr (VI) drinking and DV infection led to reduction in the weight of the spleen, but the peak reduction was seen in Cr (VI)-fed mice infected with DV, being 30, 34, and 61% at 3, 6, and 9 weeks respectively. A similar response was seen with respect to the cytotoxic activity of spleen homogenates, phagocytic activity of macrophages, and the mitogenic response of spleen cells to concanavalin A from different groups of animals, being most marked (58 to 60%) at the ninth week of Cr (VI) drinking. This shows a summation of adverse effects of DV infection in mice preexposed to Cr (VI). PMID- 20021052 TI - A comparative study on rat intestinal epithelial cells and resident gut bacteria: (I) effect of hexavalent chromium. AB - Toxicants including heavy metals reaching the intestine following ingestion through food and water primarily interact with an ecosystem of eukaryotic and prokaryotic cells. Gut bacteria having a dynamic interrelationship with intestinal epithelial cells are known to play important and specific metabolic, trophic, and protective functions. The present study was undertaken to compare the effects of hexavalent chromium on rat intestinal epithelial cells and the resident gut bacteria following in vitro and in vivo exposures. The survival rate and viability pattern of two types of cells were comparable. Under in vitro conditions, the gut bacteria were quick to reduce Cr (VI) in early time periods, while, at 30 h time, both types of cells showed similar capacity for the reduction of Cr (VI). Chromium intoxication (10 ppm of Cr (VI) in drinking water for 10 weeks) caused significant decrease in membrane alkaline phosphatase and Ca(2 +)-Mg(2 +)-ATPase activities of intestinal epithelial cells as well as of three gut bacteria viz. Escherichia coli, Pseudomonas sp, and Lactobacillus sp. Major structural membrane constituents like carbohydrates and phospholipids also showed significant decline in both types of cells. These findings indicate that 10 ppm and higher Cr concentrations may cause toxic insult, resulting in impaired intestinal functional efficacy. It also implies that the gut bacteria can be used at least for preliminary screening of heavy metals gastrointestinal toxicity. PMID- 20021053 TI - Ciprofloxacin-Induced Cytotoxicity and Apoptosis in HeLa Cells. AB - The aim of this study was to investigate the possible cytotoxic and apoptotic effects of a fluoroquinolone (FQ) antibiotic, ciprofloxacin (CPFX), on HeLa and HeLa-tat cell lines. The cultured cells were incubated with three different (20 to 100 mg/L) concentrations of CPFX, and cytotoxicity was determined by trypan blue test. A dose- and time-dependent decrease in cell proliferation was observed for both cell types except 50 to 100 mg/L CPFX for 24 h for HeLa-tat cells. CPFX induced apoptosis was also measured by the acridin orange-ethidium bromide staining using a fluorescence microscope in both cell types. Our results showed an induction of apoptosis at 100 mg/L concentration of CPFX after 24 and 48 h of incubation in both cell types. These data confirmed our previous studies obtained with normal human fibroblast cell cultures and indicated that CPFX induces cytotoxicity and apoptosis in HeLa cells. PMID- 20021054 TI - Differential regulation of free radicals (reactive oxygen and nitrogen species) by contact allergens and irritants in human keratinocyte cell line. AB - Immune responses to chemicals resulting in sensitization and the appearance of allergic responses following subsequent exposures are dependent upon activation of T lymphocytes. On the contrary, irritant responses are independent of immune response. The aim of this project was to identify the differential signaling cascade operated in allergic and irritant contact dermatitis. Recently, we have shown that keratinocyte cell line A431 can function as an antigen presenting cell (APC) and hence can be used as a model to differentiate between an allergen and irritant molecule. Allergen- and irritant-induced regulation of reactive oxygen species (ROS) and nitric oxide (NO) has been explored. Irritants induce release of ROS even at noncytotoxic concentration. ROS generation by allergens was not detected at nontoxic concentration but as the concentration was increased to a toxic dose there was a drastic increase in the ROS level compared to the untreated cells. Hence, the regulation of ROS is not significant in allergic responses but important in irritant responses. The major difference exists in the fact that the source of ROS for irritants is mitochondria, while that of allergens is mostly cytosolic. Antioxidant-induced protection from irritant induced cell death has also been demonstrated. NO level was found to increase by allergens and irritants in a concentration-dependent manner. Hence, the regulation of ROS and NO can be used as important mediators in contact allergic and irritant dermatitis. PMID- 20021055 TI - Exposure of Rosy Barb (Puntius conchonius) Sperm to Abamectin as an In Vitro Assay of Cytotoxicity. AB - The acute cytotoxicity of abamectin to the sperm of rosy barb (Puntius conchonius) was tested. It was found that 79% sperm remained motile at 30 min after activation by hypo-osmotic sodium buffer (HSB), while the motility of sperm exposed to 0.14, 0.28, and 0.42 mu M abamectin for 30 min was reduced to 60%, 35%, and 15%, respectively. Exposure to abamectin at 0.56 mu M decreased the motility of sperm to an even lower value of 5.67% after 20 min of exposure. This indicates that the toxicity of abamectin to rosy barb sperm is both dose dependent and exposure time dependent. The ultrastructural examination showed that most of the sperm exposed to 0.14 mu M abamectin for 30 min remained morphologically normal, while the midpiece and its surrounding mitochondria of the sperm treated with 0.28 mu M abamectin for 30 min were severely damaged. For the sperm treated with 0.42 mu M abamectin for 30 min, the majority of the sperm were abnormal. Sperm heads were dissolved and the midpiece broken. The mitochondria are located in the midpiece of the sperm and may be the target of abamectin toxicity in rosy barb sperm. The results also suggest that rosy barb sperm are a suitable candidate for rapid screening of the acute toxicities of bioorganic pesticides. PMID- 20021056 TI - Chemoprevention of colon cancer by a synthetic curcumin analog involves amelioration of oxidative stress. AB - The modulating effects of a bisdemethoxycurcumin analog (BDMC-A) on 1,2 dimethylhydrazine (DMH)-induced oxidative stress during colon carcinogenesis was investigated in male Wistar rats. The effects were compared with those of curcumin, a known anticarcinogen. All the animals given a weekly subcutaneous injection of DMH (20 mg/kg body wt.) for 15 weeks developed colon tumors. The colon and intestine administered DMH showed a decrease in lipid peroxidation with a concomitant increase in the activities of GSH-dependent enzymes (glutathione peroxidase, glutathione S-transferase) when compared to untreated control rats. In groups of animals given DMH and BDMC-A no tumors were observed, and the lipid peroxidation as well as the GSH-dependent enzymes showed a pattern similar to that of untreated control rats. We speculate that BDMC-A modulates DMH-induced oxidative stress and offers chemoprevention against colon carcinogenesis, and the modulatory effect is comparable with that of curcumin. Thus, lipid peroxidation and antioxidant status together could be used as markers of colon cancer chemoprevention by BDMC-A. PMID- 20021057 TI - Molecular structure-based prediction of the steady-state blood concentrations of inhaled organics in rats. AB - Chronic exposure to volatile organic chemicals (VOCs) in the environment leads to steady-state conditions. The establishment of quantitative relationships between steady-state blood concentrations and molecular structures of VOCs can be potentially useful. The objective of this study was therefore to investigate the relationship between the steady-state arterial blood concentration (Ca(ss)) in the rat and the molecular structures of 19 VOCs belonging to multiple chemical families (alkanes, haloalkanes, haloalkenes, and aromatics). The overall approach consisted of developing quantitative relationships between molecular fragments (CH(3), CH(2), CH, C, C horizontal lineC, H, Cl, benzene ring, and H in benzene ring structure) in alkanes, haloalkanes, haloethylenes, and aromatic hydrocarbons, as well as their Ca(ss) (associated with 1 mu mu mol/L inhalation exposure) according to an additive fragment model. This modeling approach implies that each fragment in the structure of a chemical has an additive and constant contribution to its Ca(ss). A multilinear regression was performed using a commercially available statistical software package, and the results obtained were essentially the contributions associated with each of the nine structural fragments toward Ca(ss) in the rat continuously exposed to 1 mu mu mol/L VOC in the air. The resulting model estimated adequately the Ca(ss) of VOCs initially used in the calibration (estimated/experimental ratio: 1.04 +/- 0.30, mean +/- standard deviation [SD]). This molecular structure vs. Ca(ss) relationship was then evaluated using an external dataset on Ca(ss) for three aliphatic hydrocarbons (octane, 2-methyl octane, and 1-nonene; 100 ppm exposures). The ratio of predicted to experimental Ca(ss) for these chemicals ranged from 0.6 to 1.2. The results of this study suggest that steady-state blood concentrations of inhaled VOCs can be predicted using structure-activity type models. PMID- 20021058 TI - A Test Procedure for the Determination of (2-Methoxyethoxy)acetic Acid in Urine from Jet Fuel-Exposed Mice. AB - A test procedure for the determination of (2-methoxyethoxy)acetic acid (MEAA) was adapted and applied to urine samples from jet fuel (JP-8)-exposed mice using capillary gas chromatography with a mass selective detector (MSD). MEAA is a metabolite and proposed biomarker for exposure to 2-(2-methoxyethoxy)ethanol, a glycol ether component in the formulation of JP-8. The collected urine samples were spiked with deuterated butoxyacetic acid internal standard, and extracted with ethyl acetate, and esterified with ethanol and sulfuric acid, and the esters of the glycol ethers were extracted with methylene chloride. The chromatographic conditions used easily separate the MEAA ethyl ester from interferences within mouse urine. The application of this procedure to urine samples collected from mice demonstrated that MEAA was detectable after oral (2000 mg/kg) or dermal (50 mu L) exposure for 7 days to JP-8 at levels as high as 8.5 or 6.5 mu g/mL, respectively. This pilot demonstration indicated that total urinary MEAA was a viable biomarker for the two routes of JP-8 exposure in laboratory mice. PMID- 20021059 TI - Ferulic Acid modulates altered lipid profiles and prooxidant/antioxidant status in circulation during nicotine-induced toxicity: a dose-dependent study. AB - Nicotine, an active ingredient of tobacco smoke, is known to induce hyperlipidemia and disturb the prooxidant-antioxidant status. In our study, ferulic acid, a naturally occurring nutritional compound, was tested for its antioxidant and antihyperlipidemic property in a dose-dependent manner against nicotine-induced toxicity in female Wistar rats. We tested three different doses of ferulic acid-10 mg, 20 mg, and 40 mg/kg body weight-for their protective effects. The activities of biochemical marker enzymes lactate dehydrogenase and alkaline phosphatase, levels of peroxidative indices (thiobarbituric acid reactive substances and hydroperoxides), nitric oxide, and circulatory lipids (cholesterol, triglycerides, free fatty acids, and phospholipids) were increased significantly in the nicotine-treated group when compared to normal, which were brought down in ferulic acid-treated groups. The antioxidant status (superoxide dismutase, catalase, glutathione peroxidase, vitamin E, and reduced glutathione) was found to be decreased in the nicotine-treated group, and was significantly increased in ferulic acid-administered groups. Further, ferulic acid also positively modulated the nicotine-induced changes in the micronutrients (zinc and copper) level. The dose 20 mg/kg body weight was found to be more effective than the other two doses. Our data suggest that FA exerts its preventive effects by modulating the degree of lipid peroxidation, antioxidant status, lipid profiles, and trace element levels during nicotine-induced toxicity. PMID- 20021060 TI - Plasma butyrylcholinesterase activity protects against cocaine hepatotoxicity in female mice. AB - Oral cocaine administration results in hepatic necrosis, increased plasma transaminase concentration, and decreased antioxidative capability, which is potentiated by lipopolysaccharide (LPS) in male CF-1 mice. Females administered the same treatment regimen display none of the hepatotoxic effects seen in their male counterparts. This study was conducted to further dissect the mechanism responsible for this gender difference in cocaine hepatotoxicity (CH) and lipopolysaccharide potentiation of CH. Male and female CF-1 mice were orally administered 20 mg/kg cocaine hydrochloride once daily for 7 days. Four hours after the last cocaine administration the mice were administered 12 x 10(6) EU LPS intraperitoneally. The activity of plasma esterase (butyrylcholinesterase), the enzyme responsible for the major pathway of cocaine metabolism to nonhepatotoxic metabolites, was measured. Aminotransferase release and histological analysis were used to determine hepatotoxicity. The concentration of the hepatotoxic precursor norcocaine was measured in the plasma and liver. Regardless of treatment, males were shown to have only 30% of the plasma esterase activity displayed by females. In addition, administration of testosterone to ovariectomized females resulted in a 70% reduction in plasma esterase activity when compared with surgically unaltered females. Moreover, hepatic norcocaine was not detected in the plasma or liver of surgically unaltered female animals, while it was present in males and testosterone-supplemented ovariectomized females. These results indicate that plasma esterase activity is heavily influenced by sex hormones, predominantly testosterone, in CF-1 mice. Suppression of plasma esterase by testosterone correlates with decreased norcocaine production and is therefore responsible, in part, for the increased CH seen following oral administration with and without LPS exposure in male CF-1 mice. PMID- 20021061 TI - Detecting role of apoptosis in mediating cyclophosphamide induced teratogenesis in vitro. AB - Programmed cell death (apoptosis) refers to a specific type of cell death under stringent genetic control. Even a slight alteration in this process leads to malformations characterized by birth defects. Based on the above hypothesis we deduced that apoptosis plays an important role in mediating the teratogenicity of cyclophosphamide in vitro. The present study was undertaken to see whether this phenomenon holds true or not. In this study, 11-day-old rat embryos were cultured for 24 hours with various concentrations of CP (i.e. 0, 5, 10 and 100 mug/ml culture). After culturing for 24 hours, embryos exposed to 10 and 100 mug/mL culture of CP were found having both malformations and growth retardation. Exposure to CP at 5 mug/mL culture did not show significant effect on embryonic development. Parallel to this, flow cytometric analysis (cell cycle and annexin V binding) and DNA fragmentation assay were also carried out followed by quantitation by 3'-OH labeling of cultured embryos to evaluate CP-induced apoptosis. All the results were found to be dose-dependant, and the data suggested that apoptosis is involved in mediating the teratogenicity of CP in vitro. PMID- 20021062 TI - Effects of cypermethrin on the dopaminergic neurons in the progressive hemiparkinsonian rats. AB - Cypermethrin is a potent pesticide derived from natural pyrethrin of the chrysanthemum plant. Cypermethrin has been known to modulate the blood-brain barrier and induce oxidative stress in rats. The oxidative stresses leading to increased reactive oxygen species generation have been identified within the degeneration of the dopaminergic (DA) neuron. However, in testing cypermethrin for its relationship to the degeneration of DA neurons, an experimental study has not yet been done. This study was designed to investigate the effects of cypermethrin on the DA neurons in the substantia nigra of normal and progressive hemiparkinsonian rats. The degree of degeneration of DA neurons was evaluated by tyrosine hydroxylase (TH) immunohistochemistry and forepaw adjusting step (FAS) test. The administration of cypermethrin (15 and 75 mg/kg/day) to the normal rats for 15 days did not decrease the number of TH-immunopositive (TH-IP) DA neurons in the substantia nigra. However, the low dose (15 mg/kg/day) of cypermethrin enhanced the rate of decline of DA neurons in the substantia nigra of hemiparkinsonian rats at 10 days and 3 weeks (p < 0.05). Also, the number of FAS in cypermethrin-treated hemiparkinsonian rats was reduced more rapidly than that of cypermethrin not-treated hemiparkinsonian rats at 10 days, 3 weeks, and 6 weeks (p < 0.05). These results suggest that cypermethrin per se cannot directly induce the degeneration of DA neurons but can accelerate a toxic effect on the degeneration of DA neurons in the progressive hemiparkinsonian rats. PMID- 20021063 TI - Effects of Chlorpyrifos and Chlorpyrifos-Methyl on the Outgrowth of Axon-Like Processes, Tubulin, and GAP-43 in N2a Cells. AB - The aim of this work was to study the neurodegenerative effects of the organophosphate (OP) pesticides chlorpyrifos (CPF) and chlorpyrifos-methyl (CHM) on cultured mouse N2a neuroblastoma cells. CPF or CHM, at a subcytotoxic concentration of 3 muM, were added to the cells either at the time of the induction of cell differentiation (codifferentiation) or 16 h after the induction of differentiation (postdifferentiation). CPF and CHM were similar in inhibiting significantly the outgrowth of axon-like processes from N2a cells after only 4 h exposure under both co- and postdifferentiation exposure conditions. Densitometric scanning of Western blots of extracts of cells treated with CPF or CHM for 4 h revealed significantly decreased cross-reactivity with a monoclonal antibody recognizing the protein GAP-43 under post- but not under codifferentiation exposure conditions. Exposure to CPF or CHM for 4 h under postdifferentiation conditions also resulted in reduced fluorescence of N2a cell body staining with anti-GAP-43. Cross-reactivity of Western blots with a monoclonal antibody recognizing alpha-tubulin was not significantly affected by OP treatment. These data indicate that a disturbance in GAP-43 may be important in the retraction of axons in predifferentiated N2a cells and support the notion that the mechanisms involved in CPF-and CHM-induced inhibition of axonal outgrowth may be different under co- and postdifferentiation exposure conditions. PMID- 20021064 TI - Oxidative stress due to nickel toxicity in the liver of protein-deficient rats. AB - This study was designed to determine the oxidative stress induced by nickel sulfate in the liver in the protein-deficient rats. Nickel sulfate in the dose of 800 mg/L in drinking water was administrated to Sprauge Dawley (SD) rats as well as protein-deficient rats for a total duration of 8 weeks. The effects of nickel treatment and protein deficiency separately and in combination were studied on rat liver antioxidant defense system enzymes like catalase, glutathione peroxidase (GPX), glutathione reductase (GR), superoxide dismutase (SOD), reduced glutathione (GSH), and glutathione-S-transferase (GST), as well as on lipid peroxidation (LPO). The investigations revealed a significant increase in the activity of enzymes, which include catalase, Gpx, GR and GST, and in the levels with LPO following nickel treatment in combination with protein deficiency. On the contrary, feeding to control rats resulted in a significant depression in the levels of SOD and GSH. However, nickel treatment to normal rats caused a significant increase in the activity of enzymes catalase and GST and in the levels of LPO, whereas the levels of GSH get significantly depressed. Further, nickel treatment to protein-deficient rats did not cause any additional alteration in the status of liver antioxidants as were observed in conditions of protein deficiency. PMID- 20021065 TI - Protective effects of selenium to placental lead neurotoxicity in rat pups. AB - The neurotoxic effects of lead are age-related phenomena, which in turn are dependent on nutrition status as well as the presence of other micronutrients. The present study was designed to study the protective effects of selenium on rat pups exposed to lead. The activities of different enzymes in cerebrum and cerebellum regions were estimated following maternal lead neurotoxicity through placenta. The parental animals were exposed to different treatments for 5 weeks prior to gestation and 3 weeks during gestation; that is, animals were exposed for a total period of 8 weeks and rat pups obtained from exposed parents were allowed to survive on their mothers' milk for a period of 1 month, after which they were sacrificed. Activities of succinic dehydrogenase (SDH), acetylcholine estrase (AChE), and Na(+)/K(+)-ATPase were studied in the cerebrum and cerebellum of the rat pups at 4 weeks of age. A significant reduction in the activity of all the three enzymes-SDH, AChE and Na(+)/K(+) -ATPase-was observed in the lead treated group, except in the case of AChE, which increased in the case of cerebrum. However, changes in different enzyme activities were less pronounced in the animals that were treated simultaneously with selenium and lead. No pups were delivered to the mothers who were exposed to selenium alone for period of 8 weeks. PMID- 20021066 TI - Platelet hyperfunction as risk factor for chronic and acute coronary events. AB - Indians have a very high incidence of vascular complications, such as hypertension, atherosclerosis, coronary artery disease (CAD), and stroke, compared to any other ethnic group in the world. They also have a very high incidence of multiple metabolic diseases, including type 2 diabetes. Elevated levels of known risk factors in this population for CAD does not explain adequately the significant increase in the vascular disease burden in this population. There is some speculation that genetic predisposal of this population may account for this increased incidence in vascular diseases. Environmental toxicants may also contribute significantly to the acceleration of these complex risk promoters. Functional and structural changes in the arterial wall precede the development of clinical complications such as endothelial dysfunction, hypertension, atherosclerosis, hyperfunction of platelets, and coagulation cascade. Vascular dysfunction, therefore, is the major contributor for the pathogenesis of hypertension, atherogenesis, thrombosis, and stroke. Alterations in the balance between platelet-associated vasoconstrictors and endothelial derived vasodilators result in the vascular dysfunction. Blood platelets play a very important role in the pathogenesis of hypertension, atherogenesis, thrombosis, and stroke. These cells interact with a variety of agonists. Such interactions stimulate specific receptors and lead to the activation of intracellular effector enzymes. Ionized calcium is the primary bioregulator and a variety of signaling mechanisms modulate the cellular physiology and functions. Activated platelets promote the formation of thrombin and initiate coagulation cascade. They also interact with other circulating blood cells and facilitate inflammatory response. Little is known about the effect of environmental toxicants on vascular physiology and pathology. This is true also on their effect on the circulating blood cells. There is some evidence that oxidative stress as well as proinflammatory compounds play a role in vascular biology. In this presentation an attempt will be made to briefly review the known risk factors for CAD; the role of toxicants, eicosanoids, and inflammatory mediators on vascular biology specifically; and the role of platelets and platelet-derived biomolecules on hypertension, atherogenesis, thrombosis, and intercellular communications. Alterations in signaling pathways by environmental toxicants may increase the risk for hypertension, atherosclerosis, thrombosis, and stroke. PMID- 20021067 TI - A mouse model for the study of vascular permeability changes induced by arsenic. AB - The primary objective of our present research was to develop an animal model for the investigation of arsenic-induced vasculopathy. Epidemiological evidence indicated that, aside from cancer, cardiovascular-related diseases were probably the most prominent health concerns in arseniasis areas. Although there were many investigations on the effect of arsenic on endothelial cells, most of these studies were conducted under in vitro conditions. A good animal model for studying the effects of arsenic on vascular integrity in vivo is very much needed. We have previously developed a rat model that could be used to demonstrate vascular changes induced by arsenic in vivo. In the present report, we are introducing a new model that is even more sensitive, economical, and effective than our original rat model. Taking advantage of the characteristics of mouse ears (thin, delicate, and with easily visible blood vessels), we demonstrated the pattern and extent of vascular leakage induced by arsenic clearly, quantitatively and convincingly. With this model, we demonstrated a time dependent increase in vascular permeability induced by arsenic (a fourfold increase in vascular leakage was observed between 10-60 minutes). With this model, we were also able to demonstrate that small caliber vessels were more vulnerable than vessels of larger caliber. Thus, this animal model can provide dynamic information (from toxic effects, pathology, and functional alterations to cellular/molecular mechanisms) that cell culture techniques are unable to provide. The introduction of this model hopefully will stimulate and inspire many other new in vivo investigations on vasculopathy induced by arsenic or other chemicals in the future. PMID- 20021068 TI - Statins and modulation of oxidative stress. AB - Statins inhibit the activity of a rate-limiting enzyme in cholesterol biosynthesis, converting 3-hydroxy 3-methylglutaryl coenzyme A to mevalonate, and are widely used in the treatment of cardiovascular diseases. Statins decrease the synthesis of cholesterol and other nonsteroid isoprenoids originating from mevalonate, such as farnesyl- and geranylgeranylpyrophosphate, dolichol, and ubiquinone. Recent studies indicate that the beneficial effect of statins on cardiovascular risk also occurs in persons with normal plasma cholesterol because of the pleiotropic cholesterol-independent activities of statins. Among these effects, modulation of oxidative stress is one of the most important. Statins reduce the generation of reactive oxygen species by vascular NAD(P)H oxidase, inhibit the respiratory burst of phagocytes, antagonize the prooxidant effect of angiotensin II and endothelin-1, and increase the synthesis of vascular nitric oxide. Some statins and their metabolites posses direct free radical scavenging activity. The antioxidant effect of statins contributes to inhibition of atherogenesis, stabilization of atherosclerotic plaque, inhibition of myocardial hypertrophy and remodeling, and modulation of vascular tone. However, the prooxidant effect of statins resulting from the inhibition of ubiquinone synthesis has also been reported in some experimental models. This effect may contribute to side effects of statins, such as myopathy and hepatotoxicity. PMID- 20021069 TI - In Vivo and In Vitro Silica Induces Nuclear Factor Egr-1 Activation Mediated by ERK 1/2 in RAW264.7 Cell Line. AB - The transcription factor early growth response gene (Egr-1) is a stress response gene activated by various forms of stress. The effect of silica on transcription and expression of Egr-1 was investigated in rat lung and in RAW264.7 cells. Silica induced the expression of Egr-1 in vivo and was mainly located in alveolar macrophage cells and lung epithelial cells. Furthermore, silica induced Egr-1 mRNA and protein expression in cultured RAW264.7 cells. Immunofluorescence microscopy revealed translocation of Egr-1 to the nucleus in response to silica. The contribution of the extracellular signal-regulated kinase (ERK) pathway to the activation of Egr-1 in response to silica was examined. Exposure to silica resulted in a rapid phosphorylation of ERK 1/2 kinases in RAW264.7 cells. MAP Kinase Kinase (MEK) inhibitor U0126 prevented Egr-1 induction by silica. The results suggest that silica could induce Egr-1 activation in macrophages in vivo and in vitro and that phosphorylated ERK 1/2 may be involved in this action. PMID- 20021070 TI - Inhibition Mechanisms of Mitochondrial Permeability Transition by 4 Hydroxytamoxifen: Protection of NAD(P)H and Thiol Group Oxidation. AB - Abstract The effects of 4-hydroxytamoxifen (OHTAM) on the mitochondrial permeability transition (MPT) induced by Ca(2+) plus peroxynitrite (ONOO(-)) or phenylarsine oxide (PhAsO) were studied to clarify its mechanisms of MPT inhibition. Ca(2+) plus ONOO(-) induced mitochondrial swelling, membrane potential (Delta Psi) depolarization, and Ca(2+) release. OHTAM, when preincubated with mitochondria, prevents those events, and if added after their induction this drug promotes a time-dependent reversal of Delta Psi depolarization and Ca(2+) release associated with MPT induction, because these events are also inhibited by cyclosporine A (CyA). Preincubation with OHTAM also inhibits thiol group oxidation associated with the MPT promoted by ONOO(-) and allows the NAD(P)(+) to recover their reduced state faster and in a higher extension. The mitochondrial swelling and Ca(2+) release after MPT induction with Ca(2+) plus PhAsO are inhibited by OHTAM; similarly to CyA, the oxidation of NAD(P)H induced by this combination is also inhibited. According to these data, the MPT inhibition by OHTAM may be related to its antioxidant capacity and the binding to target protein components of the MPT, preventing the oxidation of NAD(P)H and thiol groups, an event that increases the sensitivity to MPT induction. PMID- 20021071 TI - Changes in gene and protein expression in magnetic field-treated human glioma cells. AB - Because few cancer studies have examined protein profiles and genetic regulation from a single carcinogen exposure, the objective of this study was to determine genetic change via microarray and to evaluate whether that change was a precursor to cellular protein changes. In separate but experimentally identical studies, human glioma SF767 cells were exposed for 3 h to 60-Hz magnetic fields (sham or 1.2 muT). Microarray results suggested that magnetic field treatment resulted in the up-regulation of 5 genes, whereas 25 genes were down-regulated. The mean abundance of 10 identified proteins was altered following 1.2 muT exposure relative to sham (3 increase, 7 decrease). These studies suggest a limited but complicated response in the glioma cells to the magnetic field treatment. PMID- 20021072 TI - Impact of genetic polymorphism in relation to other factors on expression and function of human drug-metabolizing p450s. AB - A major cause for unpredictable drug response is the enormous variability of drug metabolizing cytochrome P450s (CYPs) in human liver in which genetic polymorphisms, regulation of gene expression, and physiological factors, including sex, may play a role. To dissect these different factors, we established a large human liver bank with extensive clinical documentation. Recent work concentrated on CYPs 2D6, 2B6, and the 3A family. CYP2D6 expression is highly polymorphic with over 70 alleles. Using liver samples and DNA from phenotyped patients, we further elucidated the genetic basis of phenotypic differences and demonstrated a novel role of alternative splicing, leading to decreased enzyme activity. These studies further emphasize the intricacy of genetic regulation at the CYP2D6 locus. In contrast, CYP2B6, the human orthologue of the rodent phenobarbital-inducible P450 2B, is known to be inducible by a range of substances, but our recent studies also show a high degree of genetic polymorphism. However, the role of polymorphism in determining CYP2B6 expression and function is less decisive as compared with CYP2D6. Thus, with respect to the balance between genetic and nongenetic factors controlling expression, CYP2B6 appears to take an intermediate position. CYP3A4 is the major P450 of human liver and contributes critically to the metabolism of at least half of all drugs. CYP3A4 expression variability is not influenced much by genetic polymorphism. Studies in our liver bank confirm that both induction and down-regulation occur as a consequence of drug treatment. As a further major determinant of hepatic CYP3A4 expression, we found sexual dimorphism, with women expressing at least twice the amount of CYP3A4 protein than men. This surprising result explains the many pharmacokinetic findings of faster metabolism of CYP3A4 drug substrates in women. In conclusion, these data illustrate how various factors contribute to the individual CYP profile in human liver. PMID- 20021073 TI - CYP450, COX-2 and Obesity Related Renal Damage. AB - Abstract The number of obese people in the world is growing rapidly worldwide and has reached epidemic status. Obesity is often associated with the clustering of metabolic and cardiovascular risk factors that contribute to metabolic syndrome or syndrome X. Likewise, metabolic syndrome and its associated traits are major contributing factors to the increase in nephropathy and end stage renal disease. The specific factors that link the metabolic syndrome traits to the progression of nephropathy remain largely unexplored. Recent studies have demonstrated that an imbalance between cyclooxygenase-2 (COX-2) and cytochrome P450 (CYP450) arachidonic acid metabolizing enzymes in the kidney may contribute to the renal damage associated with obesity. Along these lines, COX-2 inhibition decreases renal cytokine levels and glomerular injury in obese rats. Peroxisome proliferators-activated receptors (PPARs) are transcription factors that also contribute to chronic kidney disease in obesity and metabolic syndrome. Intriguingly, interactions between PPARs and arachidonic acid metabolites could be key determinants of renal damage in metabolic syndrome patients. Therefore, there is a promising future for pharmacological agents that manipulate COX-2 and CYP450 metabolites and PPARs to treat obesity related nephropathy. PMID- 20021074 TI - Physiologically based pharmacokinetic modeling using microsoft excel and visual basic for applications. AB - Abstract Physiologically based pharmacokinetic (PBPK) models are mathematical descriptions depicting the relationship between external exposure and internal dose. These models have found great utility for interspecies extrapolation. However, specialized computer software packages, which are not widely distributed, have typically been used for model development and utilization. A few physiological models have been reported using more widely available software packages (e.g., Microsoft Excel), but these tend to include less complex processes and dose metrics. To ascertain the capability of Microsoft Excel and Visual Basis for Applications (VBA) for PBPK modeling, models for styrene, vinyl chloride, and methylene chloride were coded in Advanced Continuous Simulation Language (ACSL), Excel, and VBA, and simulation results were compared. For styrene, differences between ACSL and Excel or VBA compartment concentrations and rates of change were less than +/-7.5E-10 using the same numerical integration technique and time step. Differences using VBA fixed step or ACSL Gear's methods were generally <1.00E-03, although larger differences involving very small values were noted after exposure transitions. For vinyl chloride and methylene chloride, Excel and VBA PBPK model dose metrics differed by no more than -0.013% or -0.23%, respectively, from ACSL results. These differences are likely attributable to different step sizes rather than different numerical integration techniques. These results indicate that Microsoft Excel and VBA can be useful tools for utilizing PBPK models, and given the availability of these software programs, it is hoped that this effort will help facilitate the use and investigation of PBPK modeling. PMID- 20021075 TI - Gene regulation and molecular toxicology. AB - Abstract The study of gene expression has become a cornerstone of molecular toxicology and toxicogenomics. From a toxicological standpoint, constitutive expression levels of a gene could be just as important in determining the outcome of toxicity as the inducible expression. There are six distinct steps at which gene expression can be controlled; these are transcription, RNA processing, RNA transport, translation, mRNA degradation, and control of protein activity. While this overall paradigm of gene regulation is still valid, the complexity of genetic regulation begins mostly at the level of transcription and certain post transcriptional events. A thorough understanding of the complexity and fluidity of gene and genome structure and their regulation is an integral part in the theory and practice of molecular toxicology and toxicogenomics. The present article is an attempt to briefly summarize our understanding of gene regulation beginning from the cistron concept. Relevance of molecular toxicology to gene regulatory mechanisms has been emphasized with examples wherever appropriate. PMID- 20021076 TI - Rapid communications: antiperspirant induced DNA damage in canine cells by comet assay. AB - Abstract Millions of people around the world use antiperspirants to decrease or eliminate body odors. Most antiperspirants contain aluminum zirconium or another form of aluminum as its active ingredient. The present investigation applied Comet assay to detect if Secret Platinum for women, Old Spice for men, or Crystal Natural produced DNA damage in Madin-Darby canine kidney cells (MDCKII). This study has shown that antiperspirants cause DNA damage on a single-cell level. Additionally, our data showed us that in general, Secret Platinum for women and Old Spice for men, produced equivalent damage. Crystal Natural, marketed as being safer or less damaging, induced the most extensive damage of all three antiperspirants tested. PMID- 20021077 TI - Analysis of multiple metabolomic subsets in vitro: methodological considerations. AB - Abstract Metabolomic analysis is a technology which seeks to provide a comprehensive profile of all the metabolites present in a biological sample ( Fiehn 2002 ; Noguchi et al. 2003 ; Watkins et al. 2002 ). Metabolomics is not a new science, but emerging as an exciting application which can span the scope of biotechnology and medicine, providing metabolic profile and a complement to the genomic and proteomic data ( German et al. 2003 ). Studies have already begun to explore the effects of toxicological, pharmaceutical, nutritional, and environmental intervention and to build integrated databases of metabolite concentrations throughout biological systems, ranging from microbes to plants to human and research animal populations ( Bruskiewich et al. 2003 ; Bailey et al. 2003 ; Fiehn 2003 ; Novotna et al. 2003 ). This may provide a tool for discovering a novel pathway, or determining the relationship of these metabolite concentrations to disease, and the extent to which nutrition can modulate metabolite concentrations ( Weckwerth and Fiehn 2002 ). Metabolites are the products of enzymatic processes. Their levels can be regarded as the ultimate response of biological systems to genetic or environmental changes ( Fiehn 2002 ). While the amount of an enzyme protein in a biological system could be quantified by using specific antibodies or by measuring the mRNA responsible for their production, factors such as the availability of cofactors and coenzymes, feedback regulation, pH, compartmentalization, etc. could affect their activity, thus, in turn, affect the metabolite's (i.e., substrates and their enzymatic products) levels. There have been a number of different interpretations of "metabolites." In pharmacological and toxicological studies, metabolites could be considered as the enzymatic degradative products of drugs ( Nishikawa et al. 2003 ; Plumb et al. 2002 ; Reo 2002 ; Robosky et al. 2002 ; Slim et al. 2002 ). In other studies, metabolites could be referred to as amino acids ( Noguchi et al. 2003 ), keto acids ( Bailey et al. 2003 ), or lipids ( German et al. 2003 ), which are substrates and products of selected metabolic pathways. Noguchi et al. (2003) proposed using "subset" to indicate the type of metabolites to be studied. Thus, the amino acid subset, for example, would indicate the levels of all the amino acids in a sample and lipomic subset would indicate the levels of all metabolites involved in lipid metabolism. In biochemical, physiological, and toxicological studies, animal models are preferred. In fact, a large amount of information on metabolites balance has been accumulated for a number of pharmacological, toxicological, and pathological conditions ( Henry et al. 1985 ; Needleman et al. 1968 ; Tourtellotte et al. 1966 ; McDougal, Jr. et al. 1997 ; Yang et al. 1985 ; King et al. 1967 ; Lowry et al. 1983 ; Gupta et al. 2000 ; Gupta et al. 2001a ,b; Gupta and Dettbarn 2003). Only recently has attention been directed towards studying metabolites in cell culture. Cell culture, with intact membrane and cytoplasmic organelles that operate under well understood biochemical and molecular events, provides a simple model that is best used for studying the basic mechanisms of toxicological and pharmacological actions of xenobiotics. Other advantages of using cell culture are to lighten the heavy workload and complications involved in performing animal studies, as well as to reduce the use of animals, which is also advocated on the ground of good animal ethics. To correlate the results in vivo, the data need to be transformed taking into consideration the level of toxicant at the site of action. PMID- 20021078 TI - Toxicokinetic modeling of parathion and its metabolites in humans for the determination of biological reference values. AB - Abstract A multi-compartment kinetic model was developed to describe the kinetics of parathion and its metabolites, p-nitrophenol (p-NP) and alkyl phosphates (AP), in order to assess worker exposure and health risks. Model compartments represent body burdens and excreta of parathion and its metabolites; to minimize the number of compartments and free parameters, regrouping was carried out on the basis of the time scales of the kinetic processes involved. Burden variations in time were described mathematically by differential equations that ensure conservation of mass on a mole basis. Model parameter values were determined from statistical fits to published in vivo kinetic data in humans. Except for the dermal absorption fraction and absorption rate, which are known to be subject to wide intra- and inter-individual variability, a single set of parameter values for the internal body kinetics enabled the model to simulate accurately the available kinetic data. For dermal exposure to parathion, with a typical absorption rate of 0.085 h(-1), model simulations show that it takes 20 h to recover half of the total amounts of p-NP eventually excreted in urine and 30 h for the AP. The model can be used to estimate the dose of parathion absorbed under different exposure routes and temporal scenarios, based on measurements of amounts of metabolites accumulated in urine over given time periods. Using the above dose-excreta links and the human no-observed-effect level for parathion reported in the literature for the inhibition of cholinesterase activities, biological reference values are proposed in the form of specific amounts of urinary metabolites excreted over chosen time periods. PMID- 20021079 TI - Effects of Echis carinatus Venom on the Haemodynamy and Contractility of Vascular and Visceral Smooth Muscle of Rats. AB - Abstract Echis carinatus is considered to be one of the most dangerous snakes in the world because of its venom toxicity and high population densities in rural agriculture areas. However, little information is available on the pharmacological effects of venom from this snake on haemodynamy and vasculature smooth muscle contractility. The haemodynamic alterations induced by the intravenous administration of Echis carinatus venom was investigated in anaesthetized rats. Injection of several doses of venom (1, 3, 5, and 7 mu g kg ( 1)) decreased the arterial blood pressure. For instance, injection of 3 mu g kg( 1) of Echis carinatus venom reduced the mean arterial blood pressure from 107 +/- 4 to 71 +/- 4 mmHg (p < 0.05). Exposure of the rat isolated vas deferens preparations to Echis carinatus venom (1 mu g ml(-1)) significantly reduced the contractile responses to epinephrine (10 mu g ml(- 1)) without changing the response to KCl (50 mM). To investigate the effects of venom on peripheral vascular resistance, mesenteric bed was removed and perfused with Krebs solution. Addition of different concentrations of venom (1, 3, and 10 mu g ml(-1)) to the precontracted mesenteric bed (with phenylephrine, 5 x 10(- 4) M) decreased the contraction in a concentration-dependent manner. However, preincubation of mesenteric bed with atropine (0.1 mu M), L-NAME (10 (-5) M) or indomethacin (10( 5) M) prevented this effect of the venom. These results may suggest that the inhibitory effect of the venom on nonvascular smooth muscle (such as vas deferens) contractions is a postsynaptic phenomenon. Furthermore, it could be suggested that the cardiovascular effects of Echis carinatus venom are mediated through multiple mechanisms such as activation of muscarinic receptors as well as production of nitric oxide and prostaglandins. PMID- 20021080 TI - Current Status of Short-Term Tests for Evaluation of Genotoxicity, Mutagenicity, and Carcinogenicity of Environmental Chemicals and NCEs. AB - The advent of the industrial revolution has seen a significant increase in the number of new chemical entities (NCEs) released in the environment. It becomes imperative to check the toxic potential of NCEs to nontarget species before they are released for commercial purposes because some of these may exert genotoxicity, mutagenicity, or carcinogenicity. Exposure to such compounds produces chemical changes in DNA, which are generally repaired by the DNA repair enzymes. However, DNA damage and its fixation may occur in the form of gene mutations, chromosomal damage, and numerical chromosomal changes and recombination. This may affect the incidence of heritable mutations in man and may be transferred to the progeny or lead to the development of cancer. Hence, adequate tests on NCEs have to be undertaken for the risk assessment and hazard prediction. Compounds that are positive in tests that detect such damages have the potential to be human mutagens/carcinogens. Only long-term animal bioassays, involving lifetime studies on animals, were used earlier to classify substances as mutagens/carcinogens. These tests were cumbersome and time consuming and required a lot of facilities and personnel. Short-term tests, therefore, were brought into practice. A "battery" of three to four of these short-term tests has been proposed now by a number of regulatory authorities for the classification of compounds as mutagenic or carcinogenic. This review deals with the current status of these short-term tests. PMID- 20021081 TI - Effects of chelating agents on distribution and excretion of terbium in mice. AB - When terbium chloride (TbCl(3)) was intravenously injected into mice, terbium (Tb) was mainly distributed into the spleen, lung, and liver. Thus, the effects of five chelating agents on the distribution of Tb to the spleen, lung, and liver of mice were examined. The treatments with diethyldithiocarbamate (DDTC), N-p methoxybenzyl-D-glucamine dithiocarbamate (MeOBGD) and 2,3-dimercaptopropanol (BAL) reduced the content of Tb in the spleen. The treatments with D penicillamine (D-PEN), ethylenediaminetetraacetic acid (EDTA), and MeOBGD reduced the content of Tb in the lung. However, BAL treatment enhanced the content of Tb in the lung, indicating the redistribution of Tb to the tissue. Although the biliary excretion of Tb was significantly increased in mice treated with EDTA and MeOBGD, these increases were negligibly small, and the metal was not detected in the urine. These results indicate that well-known chelating agents such as D-PEN, EDTA, DDTC, MeOBGD, and BAL have little ability to excrete Tb into the bile and urine. Further studies are necessary to develop a new type of chelating agent to remove Tb effectively from the body. PMID- 20021082 TI - The transcriptional effects of the vesicants lewisite and sulfur mustard on human epidermal keratinocytes. AB - Sulfur mustard (SM) injury is a complex process that begins with extensive alkylation of critical cellular components and culminates in cell death and skin vesication. The mechanism of toxicity is not well understood since SM broadly alkylates cellular nucleophiles. However, two events appear integral-the formation of DNA cross-links and the release of proteases into the extracellular matrix. To identify genes directly involved in vesication, the transcriptional profile of SM was compared to the vesicant lewisite (L). Similarly, to identify genes directly involved in DNA damage, the transcriptional profile of SM was compared to the genotoxic agent cisplatin (c-Pt). Microarrays containing 7,075 sequence-verified human cDNAs were screened with mRNA from human epidermal keratinocytes treated with 200 mu M agent for 2 h. A large number of differentially expressed genes were identified, with many similarities observed between agents. Many genes not previously associated with SM and L injury were also identified, including a large percentage of unknown function. A comparison of the differential expression profiles revealed that L had the broadest and most robustly altered expression. Apoptotic transcripts were clearly evident in L but not in SM, suggesting a late stage in L injury. PMID- 20021083 TI - Safety of a Novel Botanical Extract Formula for Ameliorating Allergic rhinitis. Part II. AB - Abstract Each year more than 50 million Americans suffer from allergic rhinitis, which is a state of hypersensitivity or hyperimmunity. Basically, allergic rhinitis is symptomatically recognized as the inflammation and irritation of the nasal mucosal membranes; sneezing; stuffy/runny nose; nasal congestion; and itchy; watery, and swollen eyes; and defined as a state of hypersensitivity/ hyperimmunity caused by exposure to a particular allergen (antigen) that results in increased reactivity upon subsequent exposure. A novel polyherbal formulation (Aller-7/NR-A2) was developed for the treatment of allergic rhinitis using a unique combination of extracts from seven medicinal plants, including Phyllanthus emblica, Terminalia chebula, Terminalia bellerica, Albizia lebbeck, Piper nigrum, Zingiber officinale, and Piper longum. Earlier studies in our laboratories have demonstrated potent antihistaminic, anti-inflammatory, antispasmodic, antioxidant, and mast-cell stabilization activities of Aller-7 in addition to its efficacy in a clinical setting. A series of preliminary toxicological evaluations were also conducted in the past, which demonstrated its safety. In this study, we have conducted further safety studies on Aller-7, including acute oral, acute dermal, acute dermal irritation, eye irritation, and 90-day repeated dose toxicity studies. Acute oral toxicity of Aller-7 was found to be greater than 5,000 mg/kg body weight in both male and female rats and no mortality or toxicity was observed at this dose, while the acute dermal toxicity was found to be greater than 2,000 mg/kg body weight. In the acute dermal irritation study, the skin irritancy index was found to be 0.0, which classifies Aller-7 as a nonirritant to rabbit skin. In the acute eye irritation study, Aller-7 was found to have minimal irritancy to eyes of rabbits. In the repeated-dose 90-day oral toxicity study, Aller-7 was administered at dose levels of 100, 300, and 1,000 mg/kg rat body weight for 90 consecutive days by oral gavage. Aller-7 did not induce any significant change in the hematological parameters. No ocular abnormalities were observed. Some minor histopathological changes were observed, but did not reveal any significant treatment-related histopathological changes. The above findings revealed that the no observed adverse effect level (NOAEL) of Aller-7 is greater than 1,000 mg/kg body weight. Taken together, these studies demonstrate the broad spectrum safety of Aller-7. PMID- 20021084 TI - In Vitro Cytotoxicity of the Organotin Compound Bis-(tri-n-butyltin)oxide to FG Cells. AB - The FG cell line derived from the gills of flounder paralichthys olivaceus was used to determine the acute cytotoxic effects of the organotin compound bis-(tri n-butyltin)oxide (TBTO). Its cytotoxic effects were initially measured by three endpoint systems: neutral red (NR) uptake assay, tetrazolium (MTT) assay, and cell protein assay. Results indicated that the doses of TBTO ranging from 1.7 x 10(- 10) to 1.3 x 10(- 7) M were all toxic, and no difference in cytotoxicity was found between the three test systems. The transmission electron microscopic examination of TBTO-exposed cells revealed that their ultrastructures were markedly altered, as evidenced by dilation of mitochondria, breakdown of rough endoplasmic reticulum, nuclear membrane dissolution, and increased level of lysosomes. It is clear that the cells are highly susceptible to TBTO. This renders FG cells one of several choices for rapidly evaluating the acute toxicities of organotin compounds like TBTO. The mode of action of TBTO leading to the cytotoxicity, including the ultrastructural alteration in FG cells, is also proposed. PMID- 20021085 TI - Effects of chromium on the resident gut bacteria of rat. AB - The major nonoccupational source of chromium (Cr) for humans is through ingestion with food and water, but its effect on the gut microflora has not been studied. The present study was, therefore, undertaken to investigate the effects of chronic ingestion of potassium dichromate (chromium VI) on the resident gut bacteria of male Wistar rats. A group of rats was kept on drinking water containing 10 ppm chromium VI (Cr [VI]) (called Cr-stressed animals) and the other group was given plain water. After 10 weeks, Lactobacillus, Pseudomonas sp., and Escherichia coli were isolated from the cecum of the rats and various studies were performed. The most significant findings of the present study were the stimulation of growth of facultative gut bacteria from the Cr-stressed rats and the significant increase of growth even in the presence of lower concentrations of Cr. Furthermore, the capacity to reduce Cr (VI) was significantly decreased along with the increased tolerance of the bacteria to Cr (higher minimum inhibitory concentration [MIC] values), which was associated with the development of antibiotic resistance. The effects were most marked with the Pseudomonas sp. and least with the E. coli. The antibiotic resistance developed with the Lactobacillus may be a blessing in disguise, because the bacteria may continue to provide benefits even in patients given antibiotic therapy. The gut bacteria thus provide the first line of defense to the body by converting toxic Cr (VI) to a less toxic Cr (III) and may act as a prebiotic. PMID- 20021086 TI - Micronucleus frequencies in exfoliated urothelial cells among individuals residing near a waste water drain and using underground water resources. AB - A study was conducted to score for any genetic damage in residents of a village of the Amritsar district. The village is located near an open drain that receives industrial effluents and municipal wastes. Earlier analyses of samples from the drain and from underground water resources of the village showed the presence of heavy metals and bacteria, respectively. As the contaminants have a potential to leach into the ground water and as the residents of the village depend only on the underground water for their anthropogenic activities, it was thought pertinent to look for genetic damage in some of the residents. Urothelial cells of 50 male residents and 25 normal healthy males (not living near any drain) were analyzed for the presence of micronuclei using standard micronucleus (MN) test protocol. The mean percent frequency of micronucleated cells in the resident group was statistically significant (t-test) as compared to the value in the control group. The MN frequency also increased with increasing age of the residents and their stay in the village. A higher frequency was further observed in the subgroup using only ground water as compared to that also using tap water (but only for drinking purposes). These results indicate exposure from genotoxicants to the residents, most probably from using underground water in the light of absence of exposure to other sources. PMID- 20021087 TI - Protective Role of a Novel Curcuminoid on Alcohol and PUFA-Induced Hyperlipidemia. AB - Alcohol use is contributing to an unprecedented decline in life expectancy. It induces hyperlipidemia when taken at higher concentrations. Alcoholics usually after a heavy binge of alcohols take fried food items normally made up of polyunsaturated fatty acids (PUFAs). The combined ingestion of alcohol and PUFAs is considered to be dangerous and known to result in hyperlipidemic conditions. Previous studies have shown that curcumin, an active principle of turmeric (Curcuma longa), has antihyperlipidemic properties. So in the present work we have synthesized an analog of curcumin and tested the protective role of that synthetic curcuminoid on alcohol and thermally oxidized sunflower oil-induced hyperlipidemia. Male Albino rats of Wistar strain were used for the experimental study. Antihyperlipidemic activity of the synthetic curcuminoid was evaluated by analyzing the levels of lipids (cholesterol, triglycerides [TGs], phospholipids [PLs], and free fatty acids [FFAs]) in different tissues and histopathological changes in the liver. The results showed that the levels of cholesterol, TGs, and FFAs were increased significantly in alcohol, thermally oxidized sunflower oil (Delta PUFA), and alcohol + Delta PUFAs treated groups. Administration of synthetic curcuminoid effectively reduced these levels. The phospholipid (PL) levels, which were decreased in the liver and kidney and increased in the heart in the alcohol, Delta PUFA, and alcohol + Delta PUFA groups, were positively modulated by treatment with synthetic curcuminoid (CA). Our histopathological observations were also in correlation with the biochemical parameters. From the results obtained, we could conclude that the synthetic curcuminoid effectively protects the system against alcohol and Delta PUFA-induced hyperlipidemia and may become an effective therapeutic agent for the treatment of hyperlipidemia. PMID- 20021088 TI - Exposure of active and passive smokers to aromatic amines present in tobacco smoke. AB - Aromatic amines, toxic agents causing methemoglobinemia and human carcinogens (2 naphtylamine, 2-toluidine), are present in mainstream (MS) and sidestream (SS) tobacco smoke. Because of this fact, active and passive smokers are exposed to them. The aim of this study was to assess exposure of active and passive smokers to aromatic amines present in tobacco smoke. Nine popular Polish cigarette brands were smoked by a smoking machine. Aromatic amines were absorbed in HCl solution and, after removing a part of matrices and alkalinizing, they were extracted with n-hexane and concentrated. Analysis by GC-FID, preceded with derivatization (with trifluoroacetic anhydride) and purification by SPE (Florisil columns), was performed. Total amounts of analyzed aromatic amines (aniline, 2-,3-,4-toluidine, 2-,3-,4-ethylaniline, 2,4-dimethylaniline) in SS varied from approximately 10 to approximately 30 mug/cigarette and were several times higher than in MS. Prognostic equation was used to calculate the theoretical concentration of aromatic amines in a model room (established number of smoked cigarettes, ventilation) and to calculate the theoretical dose of aromatic amines expired by active smokers. Results were compared with environmental standards. PMID- 20021089 TI - Evaluation of newly synthesized reactivators of the brain cholinesterase inhibited by sarin nerve agent. AB - In this work in vitro evaluation of the reactivation potency of the newly synthesized reactivators for acetylcholinesterase (AChE; EC 3.1.1.7) is described. Using this method, reactivation potency of 21 potential reactivators of AChE inhibited by the nerve agent sarin has been evaluated. We have confirmed the fact that currently the most promising AChE reactivator, HI-6, is the most effective reactivator of sarin-inhibited AChE. There are only three AChE reactivators-HI-6, TO033 and TO047-able to satisfactorily reactivate sarin inhibited AChE at the concentration 10(-5) M, which is nontoxic for human use. On the other hand, there are 14 AChE reactivators, that are able to reactivate sarin inhibited AChE at the concentration 10(-3) M. However, this concentration of reactivator is probably toxic for human use. PMID- 20021090 TI - Mitochondrial function is not affected by renal morphological changes in diabetic goto-kakizaki rat. AB - Renal disease is a common complication of diabetes mellitus. The pathogenesis of diabetic nephropathy is not well understood, but hyperglycemia seems to be a crucial factor. Recent evidence indicates that the overproduction of reactive oxygen species, observed in both clinical and experimental diabetes, and mitochondrial dysfunction are key factors in pathogenic process. The objective of this investigation was to test the hypothesis of whether hyperglycemia could affect kidney morphology and mitochondrial bioenergetics as well as susceptibility to oxidative stress in 12-month-old diabetic Goto-Kakizaki (GK) rats, a model of type 2 diabetes mellitus. We observed that there were no significant differences in the kidney respiratory function and phosphorylation capacity between GK and age-matched control Wistar rats. Mitochondria from kidneys of diabetic rats were equally susceptible to in vitro oxidative damage as those from normal rats, while coenzyme Q and alpha -tocopherol concentrations were similar in both types of preparations. However, the kidney of GK rats presented in most glomerulus a capillary basement membrane thickening with mesangial widening, in evolution to segmental glomerular sclerosis, and, in some interlobular arteries, excessive deposition of PAS-positive material at the tunica intima. The results show that the mild prolonged hyperglycemia and the kidney structural changes observed in GK rats are not sufficient to cause renal dysfunction and were not associated with functional and biochemical alterations in mitochondria. PMID- 20021091 TI - Protective Effects of N-Acetylcysteine and Vitamin E Derivative U83836E on Proteins Modifications Induced by Methanol Intoxication. AB - Methanol is oxidized into the formaldehyde and formate and these processes are accompanied by free radicals' generation. Formaldehyde and free radicals induce chemical modifications of proteins, leading to changes in their structure and function. The aim of this paper has been to evaluate the effect of N acetylcysteine and vitamin E derivative U83836E on free radicals' generation and protein modifications induced during acute methanol intoxication. U83836E is an analog of alpha-tocopherol and similarly protects cells against oxidative damage. Moreover, this compound has hydrophilic properties and can be dissolved in an aqueous phase of blood and interstitial fluid, and next, membranes readily take it up. This compound belonging to the benzopyran family contains the reactive trolox ring and possesses antioxidant properties. The ESR determination indicates the increase in free radicals' signal 6 and 12 h after intoxication. Methanol ingestion causes a significant decrease in GSH level (by about 35%) and a significant increase in the lipid peroxidation product malondialdehyde (by about 25%). During methanol metabolism the aromatic amino acids of proteins are modified-the amount of carbonyl groups is increased (by about 42%) and fluorescence intensity of tryptophan is statistically decreased (by about 30%). The increase (by about 200%) in bityrosine fluorescence is also observed. Moreover, a significant decrease in free sulphydryl (by about 40%) and amino groups (by about 30%) in liver proteins is observed during intoxication. This is accompanied by the loss of lysosomal protease-cathepsin B activity (by about 25%). N-acetylcysteine (in dose 150 mg/kg body weight) and U83836E (in dose 10 mg/kg body weight) prevent free radicals' generation to a similar degree. U83836E protects membrane phospholipids against peroxidation a little stronger than N acetylcysteine (concentration of MDA is decreased by 9 to 20% in the U83836 group and by 7 to 14% in the N-acetylcysteine group compared to methanol group). However after treating methanol-intoxicated rats with N-acetylcysteine, the changes in protein modification parameters are significantly smaller than in the group receiving methanol alone and they are a little smaller than after U83836E application. These findings suggest that N-acetylcysteine and to a smaller degree U83836E protect protein from modification in methanol intoxication, which can prevent liver pathologies. PMID- 20021092 TI - Induction of oxidative stress on reproductive and metabolic organs in sodium fluoride-treated male albino rats: protective effect of testosterone and vitamin e coadministration. AB - The present study was undertaken to search out the effect of sodium fluoride, a water pollutant noted throughout the world, including India, on oxidative stress induction in reproductive tissues, sperm pellet, and metabolic tissues like the liver and kidney. The protective effects of testosterone or vitamin-E coadministration were also observed on oxidative stress in the above mentioned samples. A significant diminution was noted in the activities of catalase and peroxidase, important antioxidant enzymes in testicular tissue, sperm pellet, prostate, and epididymis in sodium fluoride-treated rats at the dose of 20 mg/kg body weight/day (the level noted in drinking water in fluoride intoxicated areas) for 30 days by oral gavage. Coadministration of testosterone by intraperitoneal injection at the dose of 40 mug/100 g body weight/alternate day, 3 hours after fluoride treatment, resulted in a significant protection in the above mentioned parameters of all these samples. Moreover, fluoride treatment also resulted a significant elevation in the level of malondialdehyde and conjugated dienes, indicators of oxidative stress, in all the above mentioned samples, which were resettled toward the control level after testosterone coadministration. Testiculo somatic, prostato-somatic, and epididymo-somatic indices were decreased significantly in the fluoride-treated group when compared to the vehicle-treated control group. Testosterone coadministration resulted in significant restoration of these indices to the control level. We also measured the above parameters for the evaluation of oxidative stress in the liver and kidney, important metabolic organs, and noted that there was also a significant elevation in malondialdehyde and conjugated dienes along with diminution in catalase and peroxidase activities in the fluoride treated-group, with respect to the vehicle treated control group. Testosterone coadministration resulted a significant protection in these parameters toward the vehicle-treated control level. There was no significant change in hepato-somatic and reno-somatic indices among fluoride-treated, testosterone coadministered, and vehicle-treated rats. Body weight of the animals among these three groups were not changed significantly. To find out the antioxidative property of testosterone compared to vitamin E, one group of fluoride-treated animals were subjected to coadministration of vitamin E at the dose of 20 mg/100 g body weight. It was noted that in reproductive organs and in metabolic organs, oxidative stress parameters were recovered toward the control level. The results of our experiment suggests that fluoride at the dose noted in drinking water in contaminated areas may induce oxidative stress in reproductive and metabolic organs that can be ameliorated significantly by testosterone or vitamin E coadministration. Moreover, as there was no significant variation in body weights among these groups, it may be predicted that this effect of fluoride on reproductive and metabolic organs is specific and is not due to general effect of fluoride. PMID- 20021093 TI - Effects of Essential Oils and Monolaurin on Staphylococcus aureus: In Vitro and In Vivo Studies. AB - The antimicrobial properties of volatile aromatic oils and medium-chain fatty acids derived from edible plants have been recognized since antiquity. To give examples, Origanum oil, used as a food-flavoring agent, possesses a broad spectrum of antimicrobial activity due, at least in part, to its high content of phenolic derivatives such as carvacrol and thymol. Similarly, lauric acid, present in heavy concentrations in coconuts, forms monolaurin in the body that can inhibit the growth of pathogenic microbes. Using Staphylococcus aureus in broth cultures and a microdilution method, comparative efficacy of Origanum oil, and a constituent carvacrol, other essential oils and monolaurin were examined. Origanum oil was the most potent of the essential oils tested and proved bactericidal in culture to two strains of Staphylococcus aureus (ATCC #14154 and #14775) at 0.25 mg/mL. In vitro, monolaurin's effects mirrored Origanum oil. The combination of both was bactericidal at the 0.125 mg/mL concentration of each. In two separate In vivo experiments, injected Staphylococcus aureus (ATCC #14775) killed all 14 untreated mice within a 1-week period. In treated mice, over one third survived for 30 days when given oral Origanum oil daily for 30 days (6/14). Fifty percent of the mice survived for 30 days when receiving daily vancomycin (7/14) and monolaurin (4/8). Over 60% of mice survived when receiving a daily combination of Origanum oil and monolaurin (5/8). Origanum oil and/or monolaurin may prove to be useful antimicrobial agents for prevention and therapy of Staphylococcus aureus infections. PMID- 20021094 TI - Human exposure to imidacloprid from dogs treated with advantage(r). AB - The objective of this investigation was twofold: (1) to determine the transferable residue of imidacloprid in gloves worn while petting experimental household dogs after the application of Advantage(R) and (2) to determine the imidacloprid residue in the dog's blood. Advantage(R) contains 9.1% imidacloprid, which controls fleas on dogs for up to 30 days. Imidacloprid produces toxicity by interacting with nicotinic receptors. Advantage(R) (364 mg imidacloprid/dog) was applied topically to six household dogs. The glove and blood samples were collected at 24 h, 72 h, and then on a weekly basis for 5 weeks post-Advantage(R) application. The glove samples were collected by petting each dog for 5 minutes while wearing a different glove per dog. The blood samples (5 mL from each dog) were collected into EDTA tubes. The imidacloprid residue was determined in the blood extracts and glove samples using RP-HPLC. The highest levels of imidacloprid residues were detected at the 24-h interval in both glove (254.16 +/ 25.49 ppm) and blood (54.06 +/- 3.00 ppb) samples. The blood imidacloprid residue was reduced by one third at the 72-h interval (18.73 +/- 2.00 ppb) and was not detected after 1 week. Imidacloprid residue in the glove samples decreased approximately one third between each collection interval. The residue of imidacloprid in the glove extract by the fourth week was very low (0.08 +/- 0.02 ppm) and not detected by the fifth week. The present findings suggest that following topical application of Advantage(R), imidacloprid residue can be detected in the dog's blood for up to 72 h, and transferable residue on the dog's coat can be detected for up to 4 weeks. Repeated chronic exposure to imidacloprid may pose possible health risks to veterinarians, veterinary technologists, dog caretakers, and owners. PMID- 20021095 TI - Vitellogenin: a review of analytical methods to detect (anti) estrogenic activity in fish. AB - Vitellogenin, the fish egg yolk precursor protein, is the most common biochemical endpoint in general use for the detection of (anti) estrogen active substances in fish and other oviparous species. This review aims to cover the major methods (both protein and nucleic acid) for vitellogenin determination. Comparisons are drawn between vitellogenin and other endpoint effect levels. This review highlights strengths and weaknesses of the various techniques, concentrating on practical considerations and problems in evaluating the data they generate. PMID- 20021096 TI - Membrane uptake kinetics of jet fuel aromatic hydrocarbons from aqueous solutions studied by a membrane-coated fiber technique. AB - The absorption of aromatic hydrocarbons from aqueous media is a critical step involved in many biological processes after occupational and environmental exposures to jet fuel. A membrane-coated fiber (MCF) technique was used to study the uptake kinetics. A flow-through system was used to provide a constant concentration for the prolonged permeation experiments. Polydimethylsiloxane (PDMS) and polyacrylate (PA) MCFs were used to study the differential absorptivity of the aromatic compounds between the two membrane materials. The equilibrium absorption amount and a kinetic parameter describing the absorption kinetics were obtained by the regression of the permeation profiles of the aromatic compounds with a mathematical model. The partition coefficients, uptake, and elimination rate constants were determined for six benzene and three naphthalene derivatives. The PDMS/water partition coefficients of the benzene and naphthalene derivatives were linearly correlated with their logK(o/w) (LogK(pdms/w) = 0.871LogK(o/w) - 0.241, R(2) = 0.995). The PA/water partition coefficients of the benzene derivatives and the naphthalene derivatives were correlated differently with their logK(o/w). The correlation equations for benzene and naphthalene derivatives were LogK(pa/w) = 0.865LogK(o/w) + 0.0045, R(2) = 0.997 and LogK(pa/w) = 0.763LogK(o/w) + 0.911, R(2) = 1.00, respectively. These results suggest that the MCF technique can detect subtle differences in molecular interactions of the two group derivatives between the two membrane/water systems and may be used to study the absorption and permeation properties of closely related compounds. Finally, the regression method is a particularly useful tool to determine partition coefficients of very lipophilic compounds. PMID- 20021097 TI - Human exposure to selamectin from dogs treated with revolution: methodological consideration for selamectin isolation. AB - This study was undertaken to determine selamectin residue in dog's blood and in gloves worn while petting dogs after Revolution application. Revolution contains the active ingredient selamectin (a semisynthetic avermectin), which controls endoparasites and ectoparasites, including adult fleas, flea eggs, ticks, heartworms, ear mites, and sarcoptic mange in dogs, for 30 days. Revolution was applied topically on a group of six adult house hold dogs (240 mg selamectin/dog). The gloves worn for 5 min while petting the dogs were collected in glass jars and the blood samples (5 mL/dog) were collected in EDTA tubes at 0 h, 24 h, and 72 h, and at 1, 2, 3, 4, and 5 weeks post-Revolution application for selamectin residue determination. At no time during the study did the dogs show any signs of toxicity, weight loss, or change in body temperature. Extracts of the blood and the gloves were analyzed for selamectin residue using RP-HPLC coupled with a UV detector (246 nm). Selamectin standard used for peak identification and quantitation was purified from Revolution. Selamectin residue was detected in the blood (10.26 +/- 1.06 ng/mL) only at 72 h post-Revolution application, probably due to its poor dermal absorption and rapid elimination from the circulation. In the glove extracts, the highest concentration of selamectin (518.90 +/- 66.80 ppm) was detected 24 h after Revolution application. Transferable residue of selamectin in gloves from dog's coat was detected at a lesser magnitude after 1 week of Revolution application, and that was followed by a further descending trend during the second, third, and fourth weeks. No selamectin residue was detected in the glove extracts after the fifth week. In spite of selamectin's binding to the sebaceous glands of the skin, gloves contained significant transferable residue. Thus, these findings suggest that repeated exposure to selamectin can pose potential health risks, especially to veterinarians, veterinary technologists, dog trainers/handlers, and pet owners. PMID- 20021098 TI - Caspase-9 and Bax/Bcl-2 regulation in ultraviolet B-induced apoptosis of murine peritoneal macrophages. AB - This study was performed to determine the effect of UVB radiation on the activation of apoptosis regulatory proteins using murine peritoneal macrophages, which are terminally, differentiated nondividing cells. UVB (100 mJ/cm2) irradiation induced apoptosis in murine peritoneal macrophages concurrent with expression of p53, Apaf-1, upregulation of Bax, downregulation of Bcl-2, activation of caspases-9, -3, -2 and DNA fragmentation. Pretreatment of macrophages with serine protease inhibitors TPCK and TLCK inhibited UVB irradiation induced apoptosis. Interestingly, caspase-9 inhibitor Z-LEHD-FMK blocked caspase-2 activation suggesting that caspase-2 activation is not due to death receptor activation but results from activation of other caspases that are dependent on caspase-9 such as caspase-3. The data showed that the regulation of the Bcl-2 family and caspase-9 might work together to activate a caspase-3 mediated apoptotic pathway following UVB irradiation of macrophages. PMID- 20021099 TI - Sulfurtransferases and cyanide detoxification in mouse liver, kidney, and brain. AB - The activity of rhodanese, 3-mercaptopyruvate sulfurtransferase (MPST) and cystathionase in mouse liver, kidney, and four brain regions: tele-, meso-, di- and rhombencephalon was studied 30 min and 2 h following a sublethal dose of cyanide (4 mg/ kg body weight) intraperitoneal injection. Simultaneously, sulfane sulfur levels and total sulfur content, a direct or indirect source of sulfur for CN(-) conversion to SCN(-), were also investigated in these tissues. In the liver this dose of cyanide seemed to impair the process of cyanide detoxification by MPST, as well as rhodanese inhibition. The effects of cyanide administration to mice proved to be totally different in the liver and kidney. In the kidney, a significant increase in the rhodanese activity was observed as early as 30 min following cyanide intoxication, and an elevated cystathionase activity after 2 h was detected. This suggests the involvement of cystathionase in cyanide detoxification in the kidney. The activity of MPST remained at the same level as in the control group. In the rhombencephalon, similarly as in the kidney, L cysteine desulfuration pathways, which generate sulfane sulfur and sulfurtransferases that transfer sulfane sulfur atoms to CN(-), seemed to play an important role as a defense system against cyanide. The stable level of sulfane sulfur and total sulfur content was accompanied in the rhombencephalon by an increased activity of MPST, cystathionase and rhodanese. In other brain regions the role of these three sulfurtransferases was not so clear and it seemed that in the telencephalon, where the total sulfur content, but not the sulfane sulfur level, was significantly increased, some sulfur-containing compounds, such as GSH and/or cysteine, appeared in response to cyanide. PMID- 20021100 TI - Protective effect of curcumin on circulatory lipid peroxidation and antioxidant status during nicotine-induced toxicity. AB - Nicotine, an active substance present in tobacco has been identified as a major risk factor for lung related diseases. In the present study, we have evaluated the protective effect of curcumin on circulatory lipid peroxidation and antioxidants in nicotine administered Wistar rats. Lung toxicity was induced by subcutaneous injection of nicotine at a dose of 2.5 mg/kg body weight (5 days a week, for 22 weeks) and curcumin (80 mg/kg body weight) was given simultaneously along with nicotine by intragastric intubation for 22 weeks. Enhanced lipid peroxidation (41.68%) in the circulation of nicotine treated animals was accompanied by a significant decrease in the levels of ascorbic acid, vitamin E, reduced glutathione, glutathione peroxidase, superoxide dismutase and catalase. Further, reduction in the levels of zinc and elevation of copper and ferritin were observed in circulation of nicotine treated rats. Administration of curcumin significantly lowered the concentrations of lipid peroxides (36%) and enhanced the antioxidant status with modulation in the levels of zinc, copper and ferritin. Our data suggest that curcumin exerts its preventive effects by modulating the degree of lipid peroxidation, antioxidant status and trace element levels. PMID- 20021101 TI - Bio-elimination of conjugated metabolites of 3-bromobenzanthrone in urine of rats and Guinea pigs. AB - The profile of urinary metabolites of 3-bromobenzanthrone (3-BBA), an extensively used anthraquinone dye intermediate, in rats and guinea pigs was investigated using HPTLC system. A total of 10 fluorescent metabolites were detected in the urine of guinea pigs as compared to 8 in rats including the parent compound 3 BBA. The elimination of metabolites increased in a dose dependent manner in rats. The Rf values of metabolites in rats were 0.14, 0.29, 0.42, 0.52, 0.58, 0.64, 0.77 and 0.91 while that in guinea pigs were 0.23, 0.26, 0.34, 0.37, 0.44, 0.54, 0.56, 0.68, 0.76 and 0.95. The urine of 3-BBA (50 mg/kg b.wt) treated guinea pigs when digested with acid showed the disappearance of metabolite 1, 2, 5 and 7 indicating these to be the conjugated metabolites. Further, digestion of urine of 3-BBA treated guinea pigs with glucuronidase showed disappearance of metabolite 5 and 7 suggesting these as glucuronide conjugates. Digestion of urine with sulfatase enzyme resulted in disappearance of metabolite 1 which could be a sulfate conjugate. Urinary metabolite 2 which was found to be present even after digestion with glucuronidase or sulfatase but disappeared following acid treatment appears to be glutathione conjugate(s) which resulted in formation of metabolite 3 and 6. These results suggest that conjugated fluorescent metabolites of 3-BBA are excreted in urine of rats and guinea pigs. PMID- 20021102 TI - Cloning and identification of two unique genes involved in UV induced apoptosis on human keratinocyte (HaCaT) cell line. AB - Differential gene regulation during UVB induced apoptosis of human keratinocyte cell line (HaCaT) has been investigated. Rapid amplification of polymorphic DNA (RAPD)-PCR was done to identify novel/unique genes in the purified apoptotic and non-apoptotic populations. Two genes were identified and cloned in pGemT vector. One of these genes (apgene-1) was upregulated in UV induced apoptotic cells and in the non apoptotic cells exposed to UV. The other gene (apgene-2) was not detected in apoptotic cells but expressed in non-apoptotic/non necrotic cells that had been exposed to UV. The presence of apgene-1 mRNA was not detected in camptothecin induced apoptotic as well as non apoptotic cells. Apgene-2 was not detected in camptothecin induced apoptotic cells but expressed in non apoptotic/non necrotic cells. This data indicates differential regulation of these two genes during UV and chemical induced apoptosis in human keratinocytes. Additionally, since apgene-2 was upregulated in the non necrotic/non apoptotic population could be involved in protection. PMID- 20021103 TI - Automated in vitro dermal absorption (AIDA): development of a cost-effective diffusion cell. AB - A new diffusion cell design is reported for conducting in vitro dermal absorption tests. This disposable cell was inexpensive to construct and reduces the risk of inter-test cross-contamination. A disadvantage was that the polymer material used adsorbed the lipophilic test compound, 14C-nonyl phenol. Data is presented for absorption of tritiated water and 14C-nonyl phenol in a dialysis membrane model, Snakeskin and for tritiated water in human skin (Kp = 6.3 +/- 0.78 cm/h x 10( 3)). Further testing is needed to evaluate the capacity of the 'Dispo' cell for use with lipophilic test chemicals. PMID- 20021104 TI - Testing the immunosuppressive effects of cyclophosphamide in the popliteal lymph node assay in the modification of graft-vs-host reaction (PLNA-GvHR) in the rat. AB - We tested the hypothesis that popliteal lymph node assay (PLNA) in the modification of graft vs host reaction (GvHR) enabled to assess immunosuppressive potential of xenobiotics. We conducted PLNA in GvHR modification under two experimental conditions using cyclophosphamide (CY). In the first experiment average lymph node weight index (IW, weight ratio of popliteal lymph node of hind footpad injected with parental lymphocytes to lymph node of footpad injected with vehicle) in rats administered intraperitoneally with CY simultaneously with, or 3 days before local GvHR induction was 84% and 42%, respectively less than average IW in external control animals injected with parental lymphocytes into one footpad and vehicle into contralateral footpad. Average lymph node lymphocyte index (IL, ratio of cell number in the lymph node undergoing GvHR to control lymph node) in the tested animals was 92% and 86%, respectively less than in controls. In the second experiment in rats injected subcutaneously into one footpad with parental lymphocytes suspended in solution of CY at the concentrations of 4.6 mM or 18.6 mM and with parental lymphocytes into contralateral footpad, average IW was 88%and 92%, respectively less than average IW in the external control animals injected with parental lymphocytes into both footpads (average IL was 84 and 91%, respectively less than in controls). Our preliminary experiments showed a strong inhibitory effect of CY in both PLNA-GvHR models used and we believe that both models may be of value in designing future protocols aiming at prediction of immunosuppressive potential of chemicals. PMID- 20021105 TI - Comparison of Basal gene expression in cultured primary rat hepatocytes and freshly isolated rat hepatocytes. AB - Cultured primary hepatocytes are one of the most suitable in vitro models for hepatic toxicological studies. Unfortunately, there is a temporal loss of liver specific function in culture that limits their utility for some applications. Plating hepatocytes on a substratum has been shown to stabilize the differentiated phenotype for short-term culture. In order to identify the substratum that best supports in vivo basal hepatocyte gene expression profiles in vitro, the gene expression profiles of primary rat hepatocytes plated on collagen I in hepatocyte maintenance medium (HMM) or hepatocyte culture medium (HCM), or on matrigel in HMM medium for 2 h, 16 h, or 72 h were compared to the expression profiles of freshly isolated rat hepatocytes using the Atlas rat stress array. After 16 h in culture, there were differences in gene expression between cultured primary hepatocytes and freshly isolated hepatocytes, but no apparent substratum effects. At 72 h, the expression of 9 genes was altered in hepatocytes plated on either substratum compared to expression in freshly isolated hepatocytes. However, there were an additional 13 genes with increased expression in hepatocytes plated on collagen I that were expressed at low or non detectable levels in freshly isolated hepatocytes or primary hepatocytes plated on matrigel. In summary, after 72 h, primary hepatocytes plated on matrigel had basal gene expression patterns more similar to patterns in freshly isolated hepatocytes than did hepatocytes cultured on collagen. In addition, culture on matrigel suppressed the expression of atypical genes in culture. These preliminary studies suggest that culture on matrigel may be preferable for longer term in vitro toxicological studies. PMID- 20021106 TI - Regulation of mitochondrial biogenesis in eukaryotic cells. AB - Mitochondria amount within a cell is modulated in response to energy demand. This involves a tight regulation of mitochondrial biogenesis and the coordinated expression of hundreds of genes, both at the nuclear and at the mitochondrial level. This review will focus on two aspects of mitochondrial biogenesis regulation: (i) In mammalian cells, physiological effectors, and the regulatory proteins that control the expression of the respiratory apparatus, will be considered, and different kinds of tissue will be addressed. (ii) In yeast, the regulation of mitochondrial biogenesis in response to growth conditions as well as the signaling pathway involved will be considered. PMID- 20021107 TI - Genotoxic Effects of Diuron Contaminated Soil on the Root Meristem Cells of Allium sativum: A Possible Mechanism of Chromosome Damage. AB - Diuron, a persistant substituted urea herbicide, was tested in the root meristem cells of Allium sativum for the possible cytogenetic effects and to compare the sensitivity with Allium cepa. Test concentrations of diuron 22.5, 45.0, and 90.0 ppm were mixed in soil and the cloves of A. sativum were placed over diuron contaminated soils. Root meristematic cells were sampled at 48 h to score Mitotic/Chromosomal aberrations and to analyze the effect on mitotic index (MI). Microscopic analyses revealed significant and dose-dependent induction of mitotic as well as chromosomal breaks. The frequency of mitotic aberrations was every time found much higher than that of chromosomal aberrations. Mild percentage of Micronucleated and Binucleated cells was observed, as MI also declined during the analysis. Based on the data of valence charge densities on the atoms of herbicide molecule and spectroscopic studies, a possible mechanism of interaction of diuron with DNA molecule for chromosomal aberrations has been proposed. PMID- 20021108 TI - Effect of nickel (ni(2+)) on primary human ovarian granulosa cells in vitro. AB - Human ovarian granulosa cells obtained from women undergoing in vitro fertilization were exposed to 15.6, 31.25, 62.5, 125, 250, 500, 1000 muM Ni(2+) for 48 h. To determine the site of action of Ni(2+), the granulosa cells were stimulated to produce progesterone (P) by using maximally stimulating amounts of human chorionic gonadotropin (0.1 IU/ml hCG) or dibutyryl cyclic adenosine monophosphate (1 mM db-cAMP). The luteinizing hormone (LH) analog hCG was chosen because resultant P production requires an intact membrane receptor and db-cAMP was used to test for post LH receptor defects caused by Ni(2+). Progesterone content of the culture medium was determined by radioimmunoassay (RIA), and viability of the cells was measured by MTT (3-(4,5-Dimethylthiazol-2-yl)-2,5 diphenyltetrazolium bromide) reduction test. Concentration-dependent depression in both hGC and db-cAMP stimulated P production was seen at 15.625 muM or higher concentration of Ni(2+), which is not cytotoxic on human ovarian granulosa cells. The viability of cells was unaffected up to 31.25 muM and decreased significantly at 62.5 muM. Our results show a dose-related depression in stimulated P production of granulosa cells at a dose that does not induce significant cytotoxic action. These data indicate that the effect of Ni(2+) on P production is not due to cytotoxicity, and the cellular site(s) of inhibitory action appears to be subsequent to the membrane receptor and production of cAMP. PMID- 20021109 TI - In-situ measurements of low-level mercury vapor exposure from dental amalgam with zeeman atomic absorption spectroscopy. AB - Alongside food, emissions from amalgam fillings are an essential contribution to man's mercury burden. Previous methods for the determination of intraoral mercury vapor (Hg degrees ) release used principally some form of preconcentration of Hg on gold (film or wool), allowing relatively few measurements with unknown precision and sensitivity at selected times. Recently available computer controlled Hg detectors operating on Zeeman atomic absorption spectroscopy (ZAAS) facilitate the direct real-time measurement of Hg degrees concentrations. It was the aim to adapt this method for a comparative investigation of emission processes from fillings in situ and from amalgam specimens in vitro. In addition to the ZAAS instrument, the apparatus consisted of a pump, magnetic valves, an electronic flow controller and a handle with a disposable mouth piece for aspiration of oral air. A programmable timer integrated the computer-controlled instrument operation and the data collection into a standard sampling protocol. A fast exponential decay of the emission was found after stimulation of amalgam specimens and of fillings in situ (halftimes 8.6 and 10.7 min). Precision was evaluated by a series of measurements on a single patient which indicated a consistently low coefficient of variation between 18% and 25%. After insertion of a few new fillings, sensitivity was high enough to detect a significant increase in emission against the background emission from the majority of old fillings. Zeeman-AAS in connection with a semi-automated sampling protocol and data storage provides precise in-situ measurements of Hg degrees emission from dental amalgam with real-time resolution. This facilitates the detailed exploration of the Hg degrees release kinetics and the applicability to large-scale studies. PMID- 20021110 TI - Image processing of hematoxylin and eosin-stained tissues for pathological evaluation. AB - Color and geometric characteristics of stained areas in histochemical slides are among the features pathologists assess to evaluate the severity of lesions. In this research, image processing techniques were used to perform objective quantification of these characteristics in images of H&E-stained spleen tissues. A segmentation algorithm was developed to isolate the areas of interest in microscopic tissue images. Image features important to pathological evaluation were then extracted. These features were used to build statistical and neural network models to predict pathologist scores. A linear regression model predicted the scores to an R(2)-value of 0.6, and a neural network model classified samples to an accuracy of 75%. The results show the usefulness of image processing as a tool for pathological evaluation. PMID- 20021111 TI - Primary cell preparation of human renal tubular cells for transcriptome analysis. AB - We initiated a toxicogenomics project using Affymetrix GeneChip((R)) HG-U133A and HG-U133B arrays harboring 45,000 probe sets representing more than 39,000 transcripts to analyze gene expression in primary cultures of human cells after exposure to chemicals that cause tissue toxicity. In order to assess the quality of the samples studied, we prepared primary human renal cortical cell cultures from surgically resected human kidney and evaluated the origin of the cells and the effects of cryopreservation. We analyzed the primary cultures using GeneChip and compared their expression patterns with those in the Novartis Research Foundation (GNF) Gene Expression Database. The comparison with the GNF database revealed that the gene expression pattern of the cultured cells was compatible with kidney cells, indicating that we had purified human renal cortical cells. Due to the purification procedure, the primary cultured cells could be a mixture of renal components; however, we identified the major population as renal proximal tubule cells by assessing gamma-GTP activity and Glut2 antigen expression. We compared gene expression in the cells before and after cryopreservation. The expression of 567 selected housekeeping genes was unchanged by cryopreservation (Pearson's correlation coefficient r = 0.980; p < 0.0001). The analysis of more than 39,000 transcripts after normalization revealed no significant changes in expression. These results indicate that our method is satisfactory for obtaining adequate primary cell cultures of renal origin and that gene expression was not significantly changed by cryopreservation. PMID- 20021112 TI - Pretreatment of human epidermal keratinocytes in vitro with ethacrynic Acid reduces sulfur mustard cytotoxicity. AB - Sulfur mustard (SM) is a potent alkylating agent, profoundly cytotoxic, and a powerful vesicant. SM reacts quite extensively with glutathione (GSH) and forms GSH conjugates, which are presumably excreted through the mercapturic acid pathway in mammals. It is unknown whether any enzymes, such as the glutathione-S transferases (GST), are involved in this detoxification of SM by the formation of conjugates. A prototypic inhibitor (ethacrynic acid, EAA) and a prototypic inducer (Oltipraz, OLT) of GSH-S-transferase, have been used as pretreatment compounds in human epidermal keratinocytes (HEK) to investigate the effect of enzyme levels on cytotoxicity following SM challenge from 50 muM to 300 muM. Pretreatment of HEK for 24 h with EAA doubled survival against 200 muM SM (36% viability in non-pretreated cells vs. 81% in EAA-pretreated cells) and quadrupled survival (17% viability in non-pretreated controls vs. 71% in EAA-pretreated cells), while OLT pretreatment had no effect on cytotoxicity at either SM dose. The role of GST in SM cytotoxicity could not be tested because of the lack of an effect on modulation of GST activities by these 2 drugs. Cellular levels of GSH were increased 250-300% over control values using EAA pretreatment, while OLT pretreatment did not lead to any increase in GSH. Pretreatment of HEK with buthionine sulfoximine (BSO), a known depleter of glutathione levels, reduced glutathione levels and increased cytotoxicity. This large increase in GSH appears to be solely responsible for the enhanced survivability of EAA-pretreated HEK. PMID- 20021113 TI - Editorial. PMID- 20021114 TI - Uncoupling Proteins in Amoeboid Eukaryotes, Acanthamoeba castellanii, and Dictyostelium discoideum. AB - Uncoupling proteins, members of the mitochondrial carrier family, are present in the mitochondrial inner membrane, and they mediate free-fatty-acid-activated, purine nucleotide-inhibited proton reuptake. Since 1999, it has been shown that uncoupling proteins are present in some eukaryotic microorganisms, including the amoeboid protozoon Acanthamoeba castellanii and the amoeboid slime mold Dictyostelium discoideum. In the mitochondria of these organisms, uncoupling protein activity is revealed by the stimulation of state 4 respiration by free fatty acids accompanied by a decrease in membrane potential. The uncoupling proteins of amoeboid eukaryotes are able to divert energy from oxidative phosphorylation. The functional connection and physiological role of uncoupling protein and alternative oxidase in Acanthamoeba and Dictyostelium species is discussed. PMID- 20021115 TI - Mitochondrial membrane potential and ATP production in primary disorders of ATP synthase. AB - Studies of fibroblasts with primary defects in mitochondrial ATP synthase (ATPase) due to heteroplasmic mtDNA mutations in the ATP6 gene, affecting protonophoric function or synthesis of subunit a, show that at high mutation loads, mitochondrial membrane potential DeltaPsi(m) at state 4 is normal, but ADP induced discharge of DeltaPsi(m) is impaired and ATP synthesis at state 3-ADP is decreased. Increased DeltaPsi(m) and low ATP synthesis is also found when the ATPase content is diminished by altered biogenesis of the enzyme complex. Irrespective of the different pathogenic mechanisms, elevated DeltaPsi(m) in primary ATPase disorders could increase mitochondrial production of reactive oxygen species and decrease energy provision. PMID- 20021116 TI - Channels of the Outer Membrane of Saccharomyces cerevisiae Mitochondria: Cooperation and Regulation. AB - The trafficking of molecules across the mitochondrial outer membrane is mediated primarily by channels. The channel of the translocase of the outer membrane (the TOM complex channel) supports protein import, whereas metabolite transport is mediated by the voltage-dependent anion channel (the VDAC), known also as mitochondrial porin. Using a simple model of Saccharomyces cerevisiae mitochondria, it is possible to study the functional relationships among the channels as well as their regulation by cellular protein. PMID- 20021117 TI - The role of mitochondria in epilepsy: implications for neurodegenerative diseases. AB - The epileptic seizures observed in a broad variety of diseases involving mitochondrial DNA (mtDNA) and central nervous system pathology strongly suggest the possible role of mitochondria in the pathomechanism of various forms of epilepsy. The mtDNA mutations in these diseases affect the functions of complexes of oxidative phosphorylation that have mitochondria-encoded subunits. Similar deficiencies of oxidative phosphorylation, in particular of Complexes I and IV, have been detected in the epileptogenic brain regions of therapy-resistant focal epilepsies, such as the hippocampal subfield CA3 in temporal lobe epilepsy with Ammon's horn sclerosis. This suggests that impaired mitochondrial function can affect the viability and excitability of hippocampal neurons because of (1) decreased production of adenosine 5'-triphosphate; (2) increased generation of reactive oxygen species; and (3) alteration of calcium homeostasis. PMID- 20021118 TI - The various sources of mitochondrial oxygen radicals: a minireview. AB - The mitochondrial respiratory chain is the main source of reactive oxygen species in the cell. The main sites of superoxide radical production are within Complexes I and III; however, Complex II, glycerol phosphate dehydrogenase and dihydroorotate dehydrogenase, are also sources of oxygen radicals. By using specific inhibitors within the complexes it is possible to obtain indications at the sites where redox components react with oxygen and in the compartments where they are released. The production of oxygen radicals is enhanced when the electron flow is lowered, and partial uncoupling is believed to be a means of preventing excess radical production. PMID- 20021119 TI - Insulin-dependent signaling: regulation by amino acids and energy. AB - Recent research has indicated that amino acids stimulate a signal-transduction pathway that is also used by insulin. Moreover, for insulin to exert its anabolic and anticatabolic effects on protein, there is an absolute requirement for amino acids. This signaling pathway becomes inhibited by adenosine monophosphate activated protein kinase when energy production falls short. In this brief review some of the experimental evidence is discussed. PMID- 20021120 TI - Mitochondrial channels permeable by calcium ions. AB - The aim of this work was to characterize inner mitochondrial membrane channels permeable to calcium cations (Ca(++)). Mitochondrial membranes isolated from rat heart were incorporated into the bilayer lipid membrane, and single Ca(++) channel currents were measured. The observed channels were selective for Ca(++) and barium cations (Ba(++)) (53 and 50 mM) over Tris(+) (113 mM), but single channel currents in most cases were noisy and did not show the typical single channel shape, as it is known, for example, in the ryanodine receptor or in inositol 1,4,5-trisphosphate (IP(3)R) Ca(++)-release channels. The most commonly observed single-channel currents, measured at the 0 mV and 53 mM Ca(++) gradient, were in the range of 1 pA or less. The channels responded to pharmacological modulators. Some of the channels were inhibited by ruthenium red and cyclosporin A, and others were modulated by ryanodine. This may indicate that the observed channels passing Ca(++) may originate from the mitochondrial Ca(++) uniporter, the permeability transition pore, and the ryanodine receptor calcium channel. PMID- 20021121 TI - Carrier and channel properties of the mitochondrial transporters: physiology and pathology? AB - The mitochondrial metabolite transporters form a protein superfamily that is known to switch from its specific carrier mechanism to a channel/pore mode. The altered carrier function probably has pathophysiological significance. Thus, the permeability transition appears to be due to the switch of the adenine nucleotide translocator to a channel/pore mode. Similarly, when there exist abnormally high fatty-acid levels, mitochondrial carriers appear to mediate the fatty-acid uncoupling. It has been proposed that carriers facilitate the translocation of the fatty acid anion, although the possibility exists that the underlying mechanism is the conversion to the pore mode. PMID- 20021122 TI - Phytanic Acid toxicity: implications for the permeability of the inner mitochondrial membrane to ions. AB - Phytanic acid (3,7,11,15-tetramethylhexadecanoic acid), a degradation product of chlorophyll formed in the intestinal tracts of ruminants, is a constituent of milk products and meat. Because it is a branched-chain fatty acid, degradation of phytanic acid is initiated in mammalian peroxisomes. In certain peroxisomal disorders such as in Refsum disease, phytanic acid accumulates dramatically throughout the body to levels that are believed to be associated with neurological and cardiac abnormalities. Here it is shown that nonesterified phytanic acid alters the permeability of the inner mitochondrial membrane by (1) increasing the conductance to hydrogen H(+); (2) promoting opening of the permeability transition pore; and (3) initiating the release of endogenous Mg(++). But contrary to nonesterified long-chain fatty acids, phytanic acid only slightly stimulates the conductance of the inner membrane to K(+) and Cl(-). These results suggest that pathologically high tissue levels of nonesterified phytanic acid increase the permeability of the inner membrane by ions through activities that are harmful for the synthesis of mitochondrial adenosine 5' triphosphate as well as for mitochondrial integrity. PMID- 20021123 TI - RNA degradation in yeast and human mitochondria. AB - RNA turnover in yeast mitochondria is controlled by the complex called degradosome, which consists of two nuclear-encoded proteins: the SUV3 gene codes for an RNA helicase and the DSS1 gene codes for an RNase. In contrast to yeast, much less is known about RNA degradation in human mitochondria. We suggest that the key enzyme involved in this process is nuclear-encoded polynucleotide phosphorylase, hPNPase. PMID- 20021124 TI - Large-Conductance Potassium Cation Channel Opener NS1619 Inhibits Cardiac Mitochondria Respiratory Chain. AB - It has recently been reported that large-conductance calcium-cation-activated potassium channels, also known as BK(Ca)-type potassium channels, are present in the inner membranes of cardiac cell mitochondria. It was also shown that the BK(Ca)-channel opener NS1619 (1,3-dihydro-1-[2-hydroxy-5-(trifluoromethyl)phenyl] 5-(trifluoromethyl)-2H-benzimidazol-2-one) protects the heart against ischemic damage. In the present study we investigated the effects of NS1619 on the function of isolated cardiac mitochondria. In particular, we examined the influence of NS1619 on mitochondrial membrane potential and mitochondrial respiration. We showed that NS1619 decreases the mitochondrial membrane potential and inhibits the mitochondrial respiratory chain. Our results demonstrate that the protection induced by this channel opener in the heart may also be caused by pharmacological preconditioning through the inhibition of the mitochondrial respiratory chain. PMID- 20021125 TI - Mitochondria and big-conductance potassium channel openers. AB - Mitochondria play a central role in energy generation within the cell. Since the discovery of potassium channels in the inner mitochondrial membrane, mitochondria have been considered an important target for potassium channel openers. The purpose of this short review is to present the recent state of our knowledge about big-conductance potassium channels (BK-type channels) recently discovered in the inner mitochondrial membrane. In addition, modulation of mitochondrial functions by the BK-type potassium channel openers are described. PMID- 20021126 TI - Mitochondria as a target of cadmium nephrotoxicity: induction of swelling and cytochrome C release. AB - Cadmium (Cd) is a potent environmental toxic compound that damages the kidney by inducing apoptosis of the proximal tubule cells. The mitochondrion is a pivotal point of the apoptotic pathway because it releases pro-apoptotic factors such as cytochrome c. Using mitochondria isolated by differential centrifugation from rat kidney cortex, we monitored swelling, which is thought to reflect the opening of a mitochondrial permeability transition pore and mitochondrial dysfunction. At Cd concentrations of 5 muM or more, rapid swelling occurred. Moreover, Cd had concentration-dependent effects on swelling induced by the permeability transition pore opener PO(4)(3-): At concentrations of 5 muM or less, PO(4)(3-) induced swelling was enhanced and cytochrome c release was increased, whereas more than 5 muM Cd significantly inhibited both PO(4)(3-)-induced swelling and cytochrome c release. Thus, Cd may directly interfere with normal mitochondrial function and may also trigger pro-apoptotic pathways in proximal tubule cells in the kidney. PMID- 20021127 TI - Fatty-Acid-induced apoptosis in ehrlich ascites tumor cells. AB - The unsaturated fatty acids oleic acid and arachidonic acid induced typical apoptosis in Ehrlich ascites tumor cells harvested at the stationary growth phase. Arachidonic acid was a more potent inducer than oleic acid. Apoptosis was induced by concentrations of oleic and arachidonic acids that produced only partial uncoupling of the oxidative phosphorylation. The first symptoms of fatty acid-induced apoptosis included externalization of phosphatidylserine in the plasma membrane, whereas massive release of cytochrome c from mitochondria to the cytosol followed later on. PMID- 20021128 TI - Clinical Heterogeneity, Tissue Distribution, and Intergenerational Segregation of mtDNA Mutation A3243G. AB - Tissue distribution and segregation and the functional consequences of heteroplasmic mitochondrial DNA mutation A3243G were studied in 30 carriers. The mutation load in hair follicles was higher in 20 patients with a broad spectrum of clinical symptoms than in 10 nonaffected carriers. The onset of the disease negatively correlated with the mutation load in blood and muscle. The activities of respiratory chain complexes in isolated muscle mitochondria did not decrease in all patients and were normal in isolated platelets. Changes in the heteroplasmy level between pairs of mothers and offspring suggest that random genetic drift is the mechanism associated with the intergenerational transmission of the A3243G mutation. In conclusion, detailed clinical investigations and mitochondrial DNA analyses in several tissues are of the highest diagnostic value for the prognosis of the disease in carriers of the A3243G mutation. PMID- 20021129 TI - Mitochondrial DNA in tumors. AB - Mitochondria, cellular powerhouses, play a central role in apoptosis. In addition, they are suspected to contribute to aging. There is also increasing evidence that abnormalities in mitochondrial function help cancer development. Changes in mitochondrial transcription and oxidative phosphorylation pathways are observed in many types of cancer. Somatic mutations and quantitative changes in mitochondrial DNA were found in tumors, suggesting that alterations in mitochondrial DNA and the resulting respiratory deficiency play a role in carcinogenesis. It was postulated they could be useful as cancer markers. Mitochondria with abnormal functions are suitable targets for anticancer drugs. PMID- 20021130 TI - Mitochondrial DNA in polish centenarians. AB - Mitochondrial haplogroups can differ in frequency of occurrence in those who are 100 years old and in the rest of the population. These differences are not universally observed; moreover, they are population-specific. We analyzed the haplogroups in 97 Polish 100-year-olds and compared them with those of more than 145 people who served as controls. No statistically significant differences were observed. In additions, we checked D-loop sequences in 31 controls and in some of the centenarians. Statistically significant differences were found for sites 73 and 152 in the D-loop. PMID- 20021131 TI - Protective Effect of EGb 761 Against Oxidative Phosphorylation of Brain Mitochondria After Anoxia/Reoxygenation In Vivo and In Vitro. AB - The aim of this study was to investigate the effects of in vivo and in vitro anoxia and reoxygenation on the oxidative phosphorylation of brain mitochondria and to study the protective effects of Ginkgo biloba extract (EGb 761). Cerebral ischemia and reperfusion induced slight decreases in respiration rates and in the efficiency of oxidative phosphorylation. Total protection of mitochondrial function was observed after chronic pretreatment of rat with EGb 761. On the contrary, in vitro anoxia and reoxygenation of isolated brain mitochondria during respiratory assay promoted important alteration in respiration rates (around 50%) and in the oxidative phosphorylation yield (-44%). Partial protection was observed after anoxia and reoxygenation in the presence of EGb 761. Such a difference between in vivo and in vitro results could be explained by an intracellular antioxidant pool that could protect mitochondria in vivo. PMID- 20021132 TI - Skin irritation potential of copper compounds. AB - Data on the dermal irritation by copper and its compounds is scant, and its irritancy has not been determined, e.g., in terms of an irritant dose ID50. Irritancy of copper can only be comparatively characterized in relation to other metal salts. A rank order for the irritancy of metal compounds can be inferred from the patch test concentrations recommended as non-irritating for the purpose of cutaneous allergy testing: potassium dichromate 0.5% in petrolatum; copper sulfate, cobalt chloride and palladium chloride ex equo: 1% in aqueous solution, and nickel sulfate: 5% in petrolatum. PMID- 20021133 TI - Involvement of fas/fas ligand in ultraviolet B-induced apoptosis of murine peritoneal macrophages. AB - The role of Fas/Fas ligand in ultraviolet B (UVB)-induced apoptosis of murine peritoneal macrophages, the terminally differentiated, non-dividing cells was investigated. UVB (100 mJ/cm(2)) irradiation induced apoptosis in macrophages concurrent with expression of Fas, Fas ligand, Fas-associated death domain (FADD), activation of caspase-8, -3 and cleavage of poly (ADP-ribose) polymerase (PARP). Pretreatment of macrophages with a p38 mitogen activated protein kinase (MAPK) inhibitor SB202190, and c-Jun N-terminal kinase (JNK) inhibitor SP600125, inhibited UVB irradiation induced Fas expression and apoptosis. Alternatively, pretreatment with MAP kinase kinase (MEK) inhibitor PD98059, and phosphatidylinositol 3-kinase (PI3-K) inhibitor wortmannin, enhanced UVB induced expression of Fas and apoptosis. Apoptosis-inducing factor (AIF) release from mitochondria and Bcl-2 downregulation is also observed during apoptosis in UVB irradiated macrophages. The data suggests that UVB-induced apoptosis is at least in part mediated by Fas/FasL system, and that MAPKs and PI3-K play an important role in the apoptotic process of macrophages exposed to UVB irradiation. PMID- 20021134 TI - Toxicity screening of metals with special reference to quantitative approach. AB - A series of metals Cr(6+), Al(3+), Cd(2+), Pb(2+), Cu(2+), Zn(2+), and Hg(2+) were tested in three systems--Microtox, Motility Test, and Growth Zone Inhibition Test. Toxicity endpoint of each metal was variable from system to system. Of the three systems, Microtox was the most sensitive system. In this system, Hg(2+) reacted as the most toxic element having EC(50) value 0.08 mg/L while Cd(2+) and Cr(6+) were least toxic with their EC(50) values 19.4 and 21.0 mg/L respectively. MEC(90) value of Motility Test was always needed more concentrations of toxicant in comparison to other systems. In comparison to Microtox, ten times more concentration of Hg(2+) (1.4 mg/L) was required to find out its MEC(90) value. Growth Zone Inhibition Test was very simple method from handling point of view. The usual practice of evaluation of toxicity screening in this system is either qualitatively or semi-qualitatively. Hence a study was designed to establish a quantitative technique, Growth Inhibition Test, as an alternative to this test using the same sensor organism B. cereus, which allows determination of MAC as well as MIC. MIC for Hg(2+) was found to be 0.03 mg/L in Growth Inhibition Test while the same element was needed more concentration (1.0 mg/L) in the case of Growth Zone Inhibition test to produce halo. However, all these systems including Growth Inhibition Test showed Hg(2+) was the most toxic element. PMID- 20021135 TI - Interaction of emodin, aloe-emodin, and rhein with human serum albumin: a fluorescence spectroscopic study. AB - The interaction of the dihydroxyanthraquinones, emodin (1), aloe-emodin (2), and rhein (3) with human serum albumin (HSA) has been studied through fluorescence spectroscopy. Quenching studies and the association constant of the anthraquinoid compounds 1, 2, and 3 in the presence of HSA were estimated. The binding and quenching studies suggest that only emodin (1) may serve as a useful fluorescence probe for structure/function studies of different emodin binding proteins. No photoinduced binding was observed after irradiation of compounds 1, 2, and 3 in presence of human serum albumin. PMID- 20021136 TI - Carbofuran-induced endocrine disruption in adult male rats. AB - The objective of this investigation was to determine the acute toxic effects of the carbamate insecticide carbofuran on the levels of endocrine hormones in the serum of male Sprague-Dawley rats. Using chemiluminescent immunoassay, the hormones determined were progesterone, cortisol, estradiol, testosterone, triidothyronine (T3), total thyroxine (total T4), and non-protein-bound thyroxine (free T4). Rats exposed to an acute dose of carbofuran (1.5 mg/kg, s.c.) showed the onset of cholinergic signs (salivation, chewing, and fine tremors) within 5-7 min. With increasing intensity, toxic signs of maximal severity (severe convulsions and fasciculations) were observed within 30-60 min, and lasted for about 2 to 3 h. Time courses of hormones for 24 h revealed significant alterations in hormone levels during 0.5 to 3 h, with the exception of estradiol at 6 h. The levels of progesterone, cortisol, and estradiol were significantly increased (1279%, 202%, and 150%, respectively), while the levels of testosterone were decreased by 88%. No significant change occurred in thyroid hormones (T3, total T4, and free T4) at any time during the time course, despite the fact that body temperature was significantly low at 1 to 2 h after carbofuran injection. Carbofuran caused a >2-fold increase in glucose during early hours of toxicity. The results suggest that an acute exposure to carbofuran may cause transient endocrine disruption, which may consequently lead to serious reproductive problems following repeated exposure. PMID- 20021137 TI - L-Cysteine Uptake is Stimulated by 1-Chloro-2,4-Dinitrobenzene in vitro in Human Erythrocytes. AB - In previous studies, the transport of dinitrophenyl-glutathione from erythrocytes has been extensively investigated. However, the effect of treatment of erythrocytes with such xenobiotics on free-SH status of cells has not been well documented. Also, the effects of N-acetyl-L-cysteine or other-SH containing compounds on glutathione conjugate transport have not been investigated. The objectives of the present study were to investigate how the presence N-acetyl-L cysteine and L-cysteine affect the free-SH status of 1-chloro-2,4-dinitrobenzene treated erythrocytes and how N-acetyl-L-cysteine or L-cysteine affects the rate of dinitrophenyl-glutathione conjugate transport form erythrocytes. Our results indicated that L-cysteine is more efficient than N-acetyl-L-cysteine in increasing the free-SH content of erythrocytes in the presence of 1-chloro-2,4 dinitrobenzene. At the end of 20 min of exposure, free-SH levels remained at 5.3 mumol/ml erythrocyte in the presence of L-cysteine. However, in the presence of N acetyl-L-cysteine the free-SH level was 2 mumol/ml erythrocyte. In the absence of 1-chloro-2,4-dinitrobenze, L-cysteine uptake by erythrocytes was not efficient compared to N-acetyl-L-cysteine. The free-SH concentrations in the presence of N acetyl-L-cysteine and L-cysteine, in this case were, 9 +/- 1 and 1.5 +/- 0.1 mumol/ml erythrocytes respectively. These results clearly suggest that 1-chloro 2,4-dinitrobenzene stimulates the L-cysteine uptake in eryhtrocytes by a mechanism not described before. Our results also indicated that 1-chloro-2,4 dinitrobenzene induced L-cysteine uptake is a Na(+) and ATP dependent process. Replacement of NaCl with LiCl decreased the L-cysteine uptake by about 5-fold and in the presence of NaF decrease in L-cysteine uptake was about 2 fold. Our results conclude the presence of an in vitro L-cysteine uptake mechanism in erythrocytes stimulated by 1-chloro-2,4-dinitrobenzene. PMID- 20021138 TI - Absence of DNA binding activity of methyl parathion and chlorpyrifos. AB - We have investigated the characteristics of binding of the organophosphorus insecticides methyl parathion and chlorpyrifos to DNA. Quantitative analysis of our data indicates that if these compounds bind to DNA at all, it is only with weak affinity. Dissociation constants would, according to our analysis, be in excess of 1 mM. Concentrations of methyl parathion and chlorpyrifos in this range are considerably greater that those expected to occur in organisms exposed to sub lethal amounts of the compounds. Furthermore, the compounds methyl paraoxon and chlorpyrifos oxon, which result from biotransformation of methyl parathion and chlorpyrifos, exhibit similar characteristics (i.e., bind not at all or with weak affinity). These data suggest that direct binding of methyl parathion and chlorpyrifos to DNA is unlikely to be a mechanism through which any genotoxic effects are produced. PMID- 20021139 TI - Comparision of Pure Nicotine and Smokeless Tobacco Extract Induced Formation of 8 OH-dG. AB - Nicotine and smokeless tobacco extract have been shown to induce oxidative stress in different experimental systems. However, the effect of nicotine and smokeless tobacco extract containing equal amounts of nicotine on 8-OH-dG formation has not been investigated. 8-OH-dG is a DNA adduct formed by free radical attack and is elevated in tumor cells. The objective of the present study was to evaluate the formation of 8-OH-dG following exposure to different concentrations of nicotine and smokeless tobacco extract containing equal amounts of nicotine. Exposure of Chinese Hamster Ovary cells to 5 and 10 mM of nicotine resulted in generation of 8-OH-dG. The observed 8-OH-dG levels at these concantrations of nicotine were 5.1 +/- 1.7 and 9.4 +/- 2.3 8-OH-dG/dG x 100.000, respectively. Smokeless tobacco extract containing 1 and 5 mM nicotine also induced generation of 8-OH-dG. The measured 8-OH-dG levels at these concentrations were 4.7 +/- 0.6 and 20.6 +/- 2.2 8-OH-dG/dG x 100.000 respectively. Exposure of cells to smokeless tobacco extract containing 10 mM nicotine resulted in cell death. Coaddition of superoxide dismutase and catalase along with nicotine reversed the 8-OH-dG generation. However, superoxide dismutase and catalase did not inhibit the 8-OH-dG generation in smokeless tobacco extract treated cells. PMID- 20021140 TI - Brain regional heterogeneity and toxicological mechanisms of organophosphates and carbamates. AB - The brain is a well-organized, yet highly complex, organ in the mammalian system. Most investigators use the whole brain, instead of a selected brain region(s), for biochemical analytes as toxicological endpoints. As a result, the obtained data is often of limited value, since their significance is compromised due to a reduced effect, and the investigators often arrive at an erroneous conclusion(s). By now, a plethora of knowledge reveals the brain regional variability for various biochemical/neurochemical determinants. This review describes the importance of brain regional heterogeneity in relation to cholinergic and noncholinergic determinants with particular reference to organophosphate (OP) and carbamate pesticides and OP nerve agents. PMID- 20021141 TI - Incorporation of pharmacokinetic and pharmacodynamic data into risk assessments. AB - Risk assessment methodologies are being updated to allow the inclusion of numerical values for variance in pharmacokinetic (PK) measures and pharmacodynamic (PD) processes related to toxicity. The key PK measures and PD processes are identified from the results of carefully conducted and adequately reported studies. In some instances, studies with humans are not possible, and so the development of data useful for human PK evaluations and on PD processes in vitro or in silico represent an alternative. These results can be integrated under physiologic, anatomic, and biochemical constraints of the intact body through physiologically based pharmacokinetic (PBPK) modeling. This manuscript presents the rational for and key considerations related to the inclusion of quantitative PK and PD data in assessing chemical risks. PMID- 20021142 TI - Dose Related Absorption of JP-8 Jet Fuel Hydrocarbons Through Porcine Skin with Quantitative Structure Permeability Relationship Analysis. AB - The effects of dosage on the percutaneous absorption of jet fuel hydrocarbons is not clear, yet is essential for human risk assessment. The present study is an ongoing approach to assess the dose-related percutaneous absorption of a number of aliphatic and aromatic hydrocarbons. The first treatment (1X) was comprised of mixtures containing undecane (4.1%), dodecane (4.7%), tridecane (4.4%), tetradecane (3%), pentadecane (1.6%), naphthalene (1.1%), and dimethyl naphthalene (1.3% of jet fuels) in hexadecane solvent using porcine skin flow through diffusion cell. Other treatments (n = 4 cells) were 2X and 5X concentrations. Perfusate samples were analyzed with gas chromatography-flame ionization detector (GC-FID) using head space solid phase micro-extraction fiber technique. We have standardized the assay to have a good linear correlation for all the tested components in media standards. Absorption parameters including diffusivity, permeability, steady state flux, and percent dose absorbed were estimated for all the tested hydrocarbons. This approach provides a baseline to access component interactions among themselves and with the diluent (solvents). A quantitative structure permeability relationship (QSPR) model was derived to predict the permeability of unknown jet fuel hydrocarbons in this solvent system by using their physicochemical parameters. Our findings suggested a dose related increase in absorption for naphthalene and dimethyl naphthalene (DMN). PMID- 20021143 TI - Effects of dimethyl sulphoxide against liver injury caused by carbon tetrachloride in rats. AB - A single oral dose of 1.25 ml kg(-1) of carbon tetrachloride (CCl(4)) was sufficient to induce significantly elevated levels of serum glutamic pyruvic transaminase (SGPT) and serum glutamic oxaloacetic transaminase (SGOT) together with signs of acute centrilobular necrosis and fatty accumulation in liver tissue. Dimethyl sulfoxide (DMSO) in different dosages (2750 mg kg(-1), 5500 mg kg(-1) and 8250 mg kg(-1); dissolved in saline) were screened for their potential activity against CCl(4)-induced liver injury in Sprague-Dawley rats. The results showed that post-administration of high dosages (5500 mg kg(-1) and 8250 mg kg( 1)) of DMSO-saline solution significantly reduced CCl(4)-induced acute elevation in the levels of SGPT and SGOT. The same result was observed in histopathological study of liver tissue. DMSO, in high doses, probably prevented CCl(4)-induced liver injury through its antioxidant, anti-inflammatory or microsomal enzyme arresting properties. PMID- 20021144 TI - Relation between heavy metals and left ventricular diastolic function in patients with coronary artery disease. AB - One of the earliest detectable abnormalities in patients with coronary artery disease is left ventricular diastolic dysfunction. Its main cause is myocardial ischaemia, but still many of other mechanisms are possible. The aim of this study was to investigate the relation of cadmium, lead, copper, zinc, and selenium with left ventricular diastolic function indices in patients with coronary artery disease. The study included 33 patients (14 with single-vessel disease and 19 with multivessel disease on angiography) who were not exposed occupationally to heavy metals. Control group consisted of 18 patients with normal coronary arteries. The blood cadmium and lead concentrations and serum zinc, selenium and copper concentrations were determined using atomic absorption spectrophotometry. Left ventricular diastolic function parameters were estimated by echo-Doppler and colour M-mode. All heavy metal concentrations were within the standard ranges in all studied groups. Patients with single-vessel disease, multivessel disease and control group did not differ in concentrations of studied metals. On univariate and multivariate analysis only cadmium significantly correlated with echocardiographic parameters of left ventricular diastolic function: peak early to peak late mitral inflow velocity ratio (r = -0.36, p < 0.05) and deceleration time of early mitral inflow (r = 0.43, p < 0.01) in patients with CAD. No significant correlations between concentrations of heavy metals and indices of left ventricular diastolic function were found in control group. The present data suggest that cadmium in concentrations even below the threshold limit value for unexposed population may contribute to development of left ventricular diastolic dysfunction in patients with CAD, potentializing the effect of myocardial ischaemia. PMID- 20021145 TI - Development of a Guinea pig model for low-dose, long-term exposure to organophosphorus nerve agents. AB - An animal dosing model and related maximum tolerated dose (MTD) were developed for repeated exposures in guinea pigs to three organophosphorus chemical warfare nerve agents (CWNA). Male animals were injected subcutaneously with sarin (GB), soman (GD) or VX once a day (Monday through Friday) for 2-, 4-, or 13-weeks. An initial 13-week study for each CWNA employed doses of vehicle (normal saline), 0.2x, 0.4x, 0.6x, and 0.8x the previously established acute LD(50). A 2-week and 4-week exposure were also performed for each agent at doses less than the 13-week MTD to verify lack of toxicity. Animals dosed daily for 13 weeks with 0.4x LD(50) of GB or GD or with 0.2x LD(50) of VX did not display signs of acute cholinergic toxicity. In animals dosed daily for either 2- or 4-weeks, the MTDs were 0.4x the acute LD(50) for all three CWNA. There were no differences among these groups and their respective vehicle controls for weight gains, body temperature, complete blood cell counts, blood chemistries, nor by histopathology. At the MTD in all groups, red blood cell cholinesterase activity one hour after the last exposure was inhibited up to 90% compared with controls. The toxicity observed with repeated doses above the MTD for up to chronic exposure durations was not significantly different from symptoms observed after acute exposure. For all three nerve agents the MTDs for subacute exposure durations can be expressed by the same constant fraction of the acute LD(50), despite differences in the absolute amount of nerve agent administered. PMID- 20021146 TI - Cross-linking interferes with assessing sulfur mustard-induced DNA damage in human peripheral blood lymphocytes using the comet assay. AB - Sulfur mustard (SM) is a blistering agent that produces DNA strand breaks. To detect SM-induced DNA single strand breaks in human peripheral blood lymphocytes (PBL), cells were exposed to various concentrations of SM (10 to 1000 muM), and the comet assay (single-cell gel electrophoresis) was performed. We observed a SM concentration- and time-dependent increase in detectable DNA damage. To test whether SM-induced DNA cross-linking inhibits DNA migration in the comet assay, PBL were exposed to a) SM alone (10 to 1000 muM), b) H(2)O(2) (0.001%), which produces DNA single strand breaks with no cross-links, or c) SM followed at 2, 4, or 6 h by H(2)O(2). With H(2)O(2) alone, a large amount of strand breakage was detected. With H(2)O(2) plus SM, detectable H(2)O(2)-induced strand breaks decreased as SM concentration increased up to 30 muM; at 30 muM and above, the response with H(2)O(2) plus SM was similar to that with SM alone. Interference with the detection of H(2)O(2)-induced DNA strand breaks appears to be SM concentration-dependent up to 30 muM, and independent of SM concentration at >/=30 muM. This is presumably due to SM-induced cross-linking. It follows that cross-linking in DNA of SM-exposed PBL also interferes with DNA migration and detection of DNA strand breaks when cells are exposed to SM alone. PMID- 20021147 TI - Erratum. PMID- 20021148 TI - Mechanism of the Synergistic Toxicity of Malononitrile and p -Nitrobenzaldehyde with Photobacterium phosphoreum. AB - In this study the mechanism of synergistic toxicity between malononitrile and p nitrobenzaldehyde was revealed by examining their toxicodynamic and toxicokinetic interactions. The investigation of the toxicodynamic interactions focused on the interactions among malononitrile, p -nitrobenzaldehyde, and luciferase, and found that malononitrile does not induce luciferase to bind p -nitrobenzaldehyde. This result suggested that the synergistic toxicity does not arise from toxicodynamic interactions. Further research on toxicokinetic interactions compared the amounts of p -nitrobenzaldehyde being transported from the water phase to the biotic phase in two circumstances--in the presence of malononitrile and in its absence. The results of this research indicated that in the presence of malononitrile, more p -nitrobenzaldehyde is transported, and it is the increased amount of p nitrobenzaldehyde that leads to the synergistic toxicity of malononitrile and p nitrobenzaldehyde. PMID- 20021149 TI - Immunochemical Assay for Satratoxin G and other Macrocyclic Trichothecenes Associated with Indoor Air Contamination by Stachybotrys chartarum. AB - Although satratoxin G (SG) is the primary macrocyclic trichothecene produced by Stachybotrys chartarum, a fungus frequently associated with outbreaks of indoor air illness, little is known about the actual quantities of this mycotoxin in suspect or normal building environments. Using antibodies produced against an SG hemisuccinate-bovine serum albumin conjugate, we devised an enzyme-linked immunosorbent assay that enabled the detection of as little as 100 pg/mL of SG and that could also detect other members of the satratoxin family. Detection by this assay of satratoxins in methanolic extracts of S. chartarum in rice cultures was comparable to that of high-performance liquid chromatography, indicating that this assay will be useful in detecting SG and other satratoxins in environmental samples. PMID- 20021150 TI - Safety of a novel botanical extract formula for ameliorating allergic rhinitis. AB - Allergic rhinitis (also known as hay fever) is the most commonly occurring immunological disorder, and it affects 40 million men, women, and children in the United States. Symptomatically, it is an inflammation and irritation of the mucous membranes that line the nose. Allergy is defined as a state of hypersensitivity or hyperimmunity caused by exposure to a particular antigen (allergen) that results in increased reactivity upon subsequent exposure. A novel botanical formulation, Aller-7/NR-A2, was developed for the treatment of allergic rhinitis; it is a combination of medicinal plant extracts from Phyllanthus emblica, Terminalia chebula, Terminalia bellerica, Albizia lebbeck, Piper nigrum, Zingiber officinale, and Piper longum. This novel formulation has demonstrated potent antihistaminic, anti-inflammatory, antispasmodic, antioxidant, and mast cell-stabilization activities. All of the doses for these toxicity studies were selected according to the guidelines of the Organization for Economic Cooperation and Development, the World Health Organization, and the Environmental Protection Agency. Acute toxicity of Aller-7 was evaluated in Swiss Albino mice at doses of 125, 250, 500, 1000, and 1500 mg/kg. After 15 days of treatment, the animals were sacrificed. No histopathological changes were observed in major vital organs. A similar study was conducted in Albino Wistar rats, which were sacrificed at the end of 15 days. No histopathological changes or toxicity was observed at up to 2 g/kg body weight. Subacute toxicity was conducted in Albino Wistar rats at a dose of 90 mg/kg body weight for 3 days, then at 180 mg/kg for the next 3 days, and then at 270 mg/kg for 3 weeks. After 28 days, the animals were sacrificed and tested; no toxicity was observed. In a subchronic toxicity study, there was no observed adverse effect level at 1 g/kg body weight in rats. In a teratological assay, at doses of 3.0 g/kg (20 times the recommended dose) and 1.8 g/kg, respectively, no visceral or skeletal anomalies were observed in the fetuses. No maternal changes were observed when Aller-7 was administered during gestation and lactation. No evidence of mutagenicity was observed at doses up to 5000 mug per plate of Aller-7 in Salmonella typhimurium cells. The present study evaluated the safety of Aller-7 by conducting several in vitro and in vivo studies. Further studies of the 90-day chronic toxicity of Aller-7 are currently in progress. PMID- 20021151 TI - A rapid colorimetric assay for sulfur mustard cytotoxicity using isolated human peripheral blood lymphocytes and keratinocytes. AB - Sulfur mustard (SM) is a potent vesicating agent that has pronounced cytotoxic effects as well as mutagenic, carcinogenic, and radiomimetic properties. Isolated human peripheral blood lymphocytes (PBLs) and human epidermal keratinocytes (HEKs) have been used as in vitro models for determining SM-induced cytotoxicity. A recently developed colorimetric assay (the CellTiter 96 AQ ueous Non radioactive Cell Proliferation Assay) was assessed using both of the in vitro models described above. Using 24- or 96-well microplates, reproducible (+/- 10%) SM dose/response curves for both types of human cells were obtained using a spectrophotometric microplate reader set at 490 nm. After a 4-h incubation time, as many as 96 sample wells could be measured within 45 s using this commonly available equipment. Multiple plates of samples can be run immediately. This technique may facilitate cytotoxicity investigations of new candidate compounds for both prophylaxis of and therapy for SM intoxication. PMID- 20021152 TI - Modulation of energy metabolism in c6 glioma cells as possible mechanism contributing to zinc neurotoxicity. AB - Elevated tissue levels of zinc (Zn) have been associated with neurodegenerative diseases such as global ischemia, seizure, and Alzheimer's. The mechanism of action of Zn in causing neuronal injury is not clear. One of the possible mechanisms is the ability of Zn to alter cellular energy metabolism. Using the C6 glioma cell as a model, the present study aimed to determined the effects of increasing concentrations of Zn on cellular energy states, as defined by the levels of adenosine 5'-triphosphate (ATP), adenosine 5'-diphosphate (ADP), and adenosine 5'-monophosphate (AMP), the total adenosine nucleotides (TAN) (TAN = ATP + ADP + AMP), and the energy charge potential (ECP = [ATP + 0.5 ADP]/TAN). Uptake of Zn was visualized by the appearance of N-(6-methoxy-8-quinolyl)-p toluene sulfonamide (TSQ)-stained fluorescent granules after a 3-h exposure to Zn in the medium. At [Zn] = 1 mM, cells appeared apoptotic. Levels of ATP and TAN decreased as the level of Zn increased. The change mirrors the increase in cell death as determined by the trypan blue exclusion test. However, when the ratio of ATP:ADP:AMP within the TAN was calculated, the percentage of ATP in the TAN increased significantly, while that of AMP decreased. The change in the relative AMP level mirrored the change in cell viability as measured by the MTT assay, which indicated a decreased in mitochondrial activity. Cellular ECP increased significantly from 0.85 +/- 0.007 to 0.92 +/- 0.04. The elevated ECP and relative ATP level, together with a significant decrease in the relative AMP level, are all indicators of inhibition of cellular metabolism. These results support the notion that acute exposure of C6 glioma cells to a high concentration of Zn might initially result in a decrease in relative AMP and an inhibition of mitochondrial activity. However, the ultimate toxic action of Zn on the C6 glioma cells appears to be due to a gradual inhibition of energy utilization, leading to cell shrinkage and apoptosis. PMID- 20021153 TI - Toxicological and metabolic consequences of methanol poisoning. AB - Methanol, when introduced into all mammals, is oxidized into formaldehyde and then into formate, mainly in the liver. Such metabolism is accompanied by the formation of free radicals. In all animals, methanol oxidation, which is relatively slow, proceeds via the same intermediary stages, usually in the liver, and various metabolic systems are involved in the process, depending on the animal species. In nonprimates, methanol is oxidized by the catalase-peroxidase system, whereas in primates, the alcohol dehydrogenase system takes the main role in methanol oxidation. The first metabolite (formaldehyde is rapidly oxidized by formaldehyde dehydrogenase) is the reduced glutathione (GSH)-dependent enzyme. Generated formic acid is metabolized into carbon dioxide with the participation of H 4 folate and two enzymes, 10-formyl H 4 folate synthetase and dehydrogenase, whereas nonprimates oxidize formate efficiently. Humans and monkeys possess low hepatic H 4 folate and 10-formyl H 4 folate dehydrogenase levels and are characterized by the accumulation of formate after methanol intoxication. The consequences of methanol metabolism and toxicity distinguish the human and monkey from lower animals. Formic acid is likely to be the cause of the metabolic acidosis and ocular toxicity in humans and monkeys, which is not observed in most lower animals. Nevertheless, chemically reactive formaldehyde and free radicals may damage most of the components of the cells of all animal species, mainly proteins and lipids. The modification of cell components results in changes in their functions. Methanol intoxication provokes a decrease in the activity and concentration of antioxidant enzymatic as well as nonenzymatic parameters, causing enhanced membrane peroxidation of phospholipids. The modification of protein structure by formaldehyde as well as by free radicals results changes in their functions, especially in the activity of proteolytic enzymes and their inhibitors, which causes disturbances in the proteolytic-antiproteolytic balance toward the proteolytics and enhances the generation of free radicals. Such a situation can lead to destructive processes because components of the proteolytic antiproteolytic system during enhanced membrane lipid peroxidation may penetrate from blood into extracellular space, and an uncontrolled proteolysis can occur. This applies particularly to extracellular matrix proteins. PMID- 20021154 TI - Toxic Interaction of Lathyrus sativus and Manganese in Guinea Pig Intestine. AB - The combined effects on the intestinal cells of guinea pigs following feeding them with lathyrus and manganese (Mn) for 90 days were studied in this investigation. Guinea pigs given Mn (4 ppm of their diets) for 90 days showed no change in either intestinal bioconstituents or marker enzymes, with the exception of gamma-glutamyl transpeptidase (GGT) and quinone reductase (QR). Exposure to a diet of 80% lathyrus only resulted in significant (p <. 05) inhibition of intestinal alkaline phosphatase (ALP), sucrase, GGT, QR, and glutathione-s transferase (GST) along with significant (p <. 05) depletion of total hexose and phospholipids. Animals given lathyrus and Mn showed a significant (p <. 05) decrease in intestinal ALP, Ca +2 Mg +2 -ATPase, sucrase, GGT, GST, and QR along with significant (p <. 05) depletion in total hexose and phospholipids and concomitant enhancement in cholesterol when compared to controls. The data clearly indicate that combined treatment with lathyrus and Mn potentiates intestinal toxicity more than does Mn or lathyrus alone. PMID- 20021155 TI - Protection against cyclophosphamide-induced renal oxidative stress by amifostine: the role of antioxidative mechanisms. AB - Cyclophosphamide, an alkylating compound used in chemotheraphy, is metabolized into active metabolites that form reactive oxygen species. Reactive oxygen species can modify the components of both healthy and neoplastic cells in circumstances of decreased antioxidative abilities. That leads to the dysfunction of organs, including the kidneys. Therefore, drugs like amifostine, which protect healthy cells against reactive oxygen species, may be applied during cyclophosphamide therapy. The aim of this study was to evaluate the influence of amifostine on the antioxidative system of the kidneys of rats that were exposed to cyclophosphamide. Intraperitoneal administration of cyclophosphamide was found to decrease the activity of the kidney's antioxidative enzymes, such as superoxide dismutase, glutathione peroxidase, gluthatione reductase, and catalase. Amifostine, however, caused an increase in the activity of these enzymes. The administration of amifostine with cyclophosphamide partially prevented changes in the activities of the examined enzymes observed after cyclophosphamide injection. Cyclophosphamide also evoked a decrease in the levels of nonenzymatic antioxidants, such as reduced gluthatione, vitamin C, and vitamin E as well as the total antioxidant status. The administration of amifostine together with cyclophosphamide prevented changes in the concentration of evaluated nonenzymatic antioxidative parameters, increasing values of their concentration to the values in the control group. Changes in the kidneys' antioxidative abilities during detoxification from cyclophosphamide were accompanied by intensified lipid peroxidation, which was manifested by an increase in the concentration of malondialdehyde and 4-hydroxynonenal. Amifostine caused the inhibition of lipid peroxidation in the kidneys of control and cyclophosphamide-treated rats. In conclusion, our results suggest that amifostine significantly protects kidney antioxidant parameters from changes caused by cyclophosphamide treatment and, in consequence, prevents oxidative stress and phospholipid peroxidative damage. Thus, amifostine prevents renal injury and dysfunction. PMID- 20021156 TI - Methodology for evaluating oxidative DNA damage and metabolic genotypes in human trabecular meshwork. AB - In vitro studies have suggested the possible role of oxidative DNA damage in degenerative eye diseases such as glaucoma. We propose a method aimed at evaluating the oxidative molecular damage directly in the human trabecular meshwork (HTM) collected during surgery from patients affected by glaucoma. In the same DNA samples, we evaluated two genes involved in the cellular defense against oxidative stress, the glutathione S -transferase-encoding genes GSTM1 and GSTT1. DNA was extracted, using a high-performance phenol/chloroform procedure, from the HTM collected during surgery from nine glaucoma patients and five controls. Oxidative DNA damage was evaluated by determining the levels of 8 hydroxy-2'-deoxyguanosine (8-OHdG) by means of 32 P postlabeling, thin-layer chromatography, and electronic autoradiography. GSTM1 and GSTT1 polymorphisms were determined by polymerase chain reaction (PCR) and agarose electrophoresis. Sufficient DNA amounts were obtained from all examined specimens. 8-OHdG was detected in all samples, with a level of 4.0 +/- 6.5 (mean +/- SD) 8-OHdG molecules/10 5 normal nucleotides in the glaucoma patients and a level of 2.6 +/- 2.2 in the controls. These results were obtained by using DNA amounts as low as 0.11 mug. The genotype status of GSTM1 and GSTT1 was successfully determined in all patients by analyzing an aliquot of the same DNA used for the 8-OHdG evaluation. This method allows for the use of samples collected from living subjects during ocular surgery in order to study the role of oxidative DNA damage in the pathogenesis of degenerative eye diseases such as glaucoma. PMID- 20021157 TI - Acrylonitrile-induced neurotoxicity in normal human astrocytes: oxidative stress and 8-hydroxy-2'-deoxyguanosine formation. AB - Studies in our laboratory and others have indicated that acrylonitrile (VCN) induces acute and chronic neurotoxicity and brain tumors in animal models. Reduced glutathione (GSH) depletion was suggested as the initiator of oxidative stress in VCN-induced neurotoxicity. Astrocytes possess the majority of total brain GSH and express various immunological functions that are characteristic of the brain, including the secretion of cytokines. We hypothesized that astrocytes could be the primary target of VCN's adverse activities in the brain. Therefore, VCN-induced neurotoxicity was studied by exposing proliferating normal human astrocytes (NHAs) to various concentrations of VCN (25-400 muM). We assessed cell viability; levels of endogenous antioxidants, GSH, and catalase; levels of reactive oxygen species; and secretion of TNF- alpha, a cellular marker for oxidative stress and oxidative damage to nuclear DNA, after treatment with VCN. At VCN concentrations of 25 and 50 muM, the oxidative stress markers were unaffected and at least 85% of the cells were viable. Cell viability was significantly affected at 200 and 400 muM VCN (22-42% less than control, p <. 05). The results also indicated VCN-induced depletion of GSH and a concomitant increase in levels of oxidized GSH (GSSG). The levels of total GSH and GSSG in control and treated (400 muM VCN) cells were 37 and 2, respectively. There was a significant upregulation of catalase activity (21% more than control, p <. 05) at 100 muM of VCN and a downregulation at 400 muM (40% lower than control, p <. 05). A dose-dependent, significant increase in the formation of reactive oxygen species was observed at 200 to 400 muM of VCN. Also, an elevation (two- to three fold as compared to control, p <. 05) in oxidative damage to DNA was observed at these concentrations of VCN. Increase in TNF- alphasecretion (28% higher than control, p <. 05) was observed at 400 muM VCN. Therefore, redox imbalance in astrocytes may play a major role in VCN-induced neurotoxicity, which is indicated by compromised antioxidant defense mechanisms, such as depletion of GSH, increase in GSSG, inhibition of catalase, and increase in the formation of reactive oxygen species and TNF- alphasecretion, resulting in DNA oxidation. PMID- 20021158 TI - Pulse Analysis as a Possible Real-Time Biomarker Complementary to SGPT and SGOT for Monitoring Acute Hepatotoxicity. AB - Based on the resonance theory, the pressure wave of the arterial system could reflect the physical condition of the internal organs. Experimental evidence indicates that the physical condition of organs is related to various Fourier components of the pressure pulse. We have proved that the first harmonic of the pressure-wave spectrum is closely related to the liver. In this study, Wistar rats given massive doses of acetaminophen were examined. The amplitude of the first harmonic of the blood pressure pulse, A1, was analyzed and correlated with the blood liver function indexes that examine serum glutamate pyruvate transaminase (SGPT) and serum glutamic oxaloacetic transaminase (SGOT). When A1 was below 3650 and SGPT was above 90, the kappa value was about 0.6; the probability is greater than. 999 based on a chi-square test. When A1 was below 3650 and SGOT was above 380, the kappa value was around 0.5; the probability is greater than. 999. Our results indicate that A1 may be used as a simple, real time biomarker that is complementary to the commonly used first-line liver indicators, SGPT and SGOT levels, for monitoring acute hepatotoxicity. It promises a noninvasive, real-time liver-function monitoring method. PMID- 20021159 TI - Lipid peroxidation-independent mechanisms of vitamin e-mediated protection against cyclosporine a-induced hepatocellular toxicity and apoptosis. AB - Cyclosporine A (CsA) produces oxidative stress and apoptosis in rat hepatocytes, but it is not known whether membrane lipid peroxidation plays a role in CsA toxicity. The objective of the present study was to determine whether iron catalyzed hydroxyl or membrane alkoxyl radical formation is causally involved in the prooxidative cell injury and apoptosis. As previously reported, cultured rat hepatocytes exposed to CsA exhibited concentration-dependent signs of apoptotic cell injury, including chromatin condensation and fragmentation, increased caspase-3 activity, and release of cytosolic lactate dehydrogenase. Addition of the ferric iron chelator desferrioxamine or the novel potent lipophilic iron chelator CGP72670 did not protect against CsA-induced cell death, indicating that iron-catalyzed lipid peroxidation is unlikely to be involved in CsA-mediated apoptosis. In contrast, the classical chain-breaking lipid peroxidation inhibitor alpha-tocopherol was able to rescue cells from CsA-induced cytotoxicity and apoptosis. alpha-Tocopherol not only effectively inhibited the production of thiobarbituric acid-reactive substances and the formation of reactive oxygen species, but it also prevented proteins from being oxidized and forming mixed disulfides. Furthermore, alpha-tocopherol inhibited the cellular uptake of extracellular calcium (45 Ca 2+) into cells, similar to the reducing agent dithiothreitol. By decreasing the extracellular calcium concentrations or by adding calcium channel blockers (diltiazem, nifedipine) or a cell-permeable calcium chelator Bis-(o-aminophenoxy)-ethane-N,N,N',N'-tertraacetic acid (BAPTA), both CsA-induced caspase-3 activation and apoptosis were inhibited, indicating a pivotal role of Ca 2+ in mediating CsA-induced cell injury. These results suggest that alpha-tocopherol protects from CsA-mediated apoptosis and cytotoxictiy by preventing the oxidation of redox-sensitive Ca 2+ -ATPase. Thus, it is the attenuation of increased calcium influx and, hence, the inhibition of caspase-3 activation, rather than the downstream inhibition of lipid peroxidation, that is a key mechanism of the protection provided by alpha-tocopherol against CsA induced cell injury. PMID- 20021160 TI - The toxicokinetics of cyanide and mandelonitrile in the horse and their relevance to the mare reproductive loss syndrome. AB - The epidemiological association between black cherry trees and mare reproductive loss syndrome has focused attention on cyanide and environmental cyanogens. This article describes the toxicokinetics of cyanide in horses and the relationships between blood cyanide concentrations and potentially adverse responses to cyanide. To identify safe and humane blood concentration limits for cyanide experiments, mares were infused with increasing doses (1-12 mg/min) of sodium cyanide for 1 h. Infusion at 12 mg/min produced clinical signs of cyanide toxicity at 38 min; these signs included increased heart rate, weakness, lack of coordination, loss of muscle tone, and respiratory and behavioral distress. Peak blood cyanide concentrations were about 2500 ng/mL; the clinical and biochemical signs of distress reversed when infusion stopped. Four horses were infused with 1 mg/min of sodium cyanide for 1 h to evaluate the distribution and elimination kinetics of cyanide. Blood cyanide concentrations peaked at 1160 ng/mL and then declined rapidly, suggesting a two-compartment, open model. The distribution (alpha) phase half-life was 0.74 h, the terminal (beta phase) half-life was 16.16 h. The mean residence time was 12.4 h, the steady-state volume of distribution was 2.21 L/kg, and the mean systemic clearance was 0.182 L/h/kg. Partitioning studies showed that blood cyanide was about 98.5% associated with the red cell fraction. No clinical signs of cyanide intoxication or distress were observed during these infusion experiments. Mandelonitrile was next administered orally at 3 mg/kg to four horses. Cyanide was rapidly available from the orally administered mandelonitrile and the C max blood concentration of 1857 ng/mL was observed at 3 min after dosing; thereafter, blood cyanide again declined rapidly, reaching 100 ng/mL by 4 h postadministration. The mean oral bioavailability of cyanide from mandelonitrile was 57% +/- 6.5 (SEM), and its apparent terminal half life was 13 h +/- 3 (SEM). No clinical signs of cyanide intoxication or distress were observed during these experiments. These data show that during acute exposure to higher doses of cyanide (~600 mg/horse; 2500 ng/mL of cyanide in blood), redistribution of cyanide rapidly terminated the acute toxic responses. Similarly, mandelonitrile rapidly delivered its cyanide content, and acute cyanide intoxications following mandelonitrile administration can also be terminated by redistribution. Rapid termination of cyanide intoxication by redistribution is consistent with and explains many of the clinical and biochemical characteristics of acute, high-dose cyanide toxicity. On the other hand, at lower concentrations (<100 ng/mL in blood), metabolic transformation of cyanide is likely the dominant mechanism of termination of action. This process is slow, with terminal half-lives ranging from 12-16 hours. The large volume of distribution and the long terminal-phase-elimination half-life of cyanide suggest different mechanisms for toxicities and termination of toxicities associated with low-level exposure to cyanide. If environmental exposure to cyanide is a factor in the cause of MRLS, then it is likely in the more subtle effects of low concentrations of cyanide on specific metabolic processes that the associations will be found. PMID- 20021161 TI - Image Processing to Facilitate Histological Evaluation of Tissue Specimens Stained With Perl's Prussian Blue. AB - Image-processing techniques were developed to assist histological analysis of rat spleen specimens stained for iron with Perl's Prussian blue. The stained areas in the splenic tissue images were segmented on the basis of color attributes. A series of image features was extracted to describe the subjective concept of blueness, an important attribute in the histological evaluation of blue-stained tissues. Feature extraction was based on statistical analysis. A neural network model was developed to predict pathologists' scores from selected image features. The model predicted pathologists' scores with an R 2 value of 0.86. PMID- 20021162 TI - Photo-induced interaction of antibacterial quinolones with human serum albumin. AB - The absorption and fluorescence spectrums of four antibacterial quinolones, namely, ciprofloxacin, norfloxacin, enoxacin, and cinoxacin, were studied in the presence of human serum albumin (HSA). Of the three fluoroquinolones studied, ciprofloxacin and norfloxacin were found to bind efficiently to HSA when irradiated with visible light, whereas the third, enoxacin, bound only moderately. On the other hand, cinoxacin, a nonfluorinated quinolone of the first generation, did not show any interaction with HSA. The findings were inferred by monitoring the evolution of the fluorescence spectrums of the solutions as a function of time. A direct relationship between the capacity of the photo-induced defluorination to produce aryl cation intermediates, and the subsequent binding reaction with HSA, was observed and is discussed. PMID- 20021163 TI - Sequential Changes in the Cellular Redox State During 7,12 Dimethylbenz[a]anthracene-Induced Carcinogenesis in the Hamster Buccal Pouch. AB - Our objective was to elucidate sequential changes in the oxidant-antioxidant status during 7,12-dimethylbenz[a]anthracene (DMBA)-induced hamster buccal pouch (HBP) carcinogenesis. In designing the study, we divided hamsters into experimental and control groups. The right buccal pouches of the experimental animals were painted three times a week with a 0.5% solution of DMBA in liquid paraffin. The control animals received paraffin alone. The hamsters were killed after 1, 4, 8, 12, and 16 weeks of DMBA treatment, and the buccal pouches were examined for histopathological changes. The extent of lipid peroxidation and the status of glutathione-dependent antioxidants were evaluated in the buccal pouch, liver, and erythrocytes. Our results showed that the experimental animals developed severe hyperplasia and hyperkeratosis after 4 weeks, dysplasia after 8 weeks, and well-developed squamous cell carcinomas after 16 weeks of DMBA application. Topical application of DMBA increased lipid peroxidation in the buccal pouch up to the 8th week; there was a substantial fall after 12 weeks and significantly low levels after 16 weeks. This was accompanied by a sustained increase in reduced glutathione (GSH) and the activities of glutathione peroxidase (GPx), glutathione-S-transferase (GST), and gamma glutamyltranspeptidase (GGT) throughout the carcinogenic progression. However, in the liver and erythrocytes, the concentrations of lipid peroxides were higher, and GSH- and GSH-dependent enzyme activities were lower than in the controls throughout the experiment. This study has revealed intrinsic differences in the cellular redox state in the target organ and host tissues of tumor-bearing animals. We suggest that measurement of lipid peroxidation and GSH-dependent antioxidants could be valuable in evaluating carcinogenic progression and the effects of putative chemopreventive agents in the hamster buccal pouch model. PMID- 20021164 TI - Nuclear magnetic resonance analysis of the solution and solvolysis of sulfur mustard in deuterium oxide. AB - Our laboratory performs in vitro experiments in which cell cultures are exposed to sulfur mustard (HD) to investigate the toxicity of this agent of chemical warfare. To perform these experiments, it is important to know the rate of hydrolysis of HD in order to calculate the concentrations of HD and its hydrolysis products during the experiment. Researchers have previously investigated the kinetics and mechanism of the hydrolysis of HD using a variety of methods. In the present study, we used nuclear magnetic resonance (NMR) spectroscopy and gas chromatography/mass spectrometry (GC/MS) to investigate HD's dissolution and solvolysis in deuterium oxide (D 2 O) at 2 mM. We followed activity in proton spectrums and determined the half-life (t 1/2) of HD to be 7.0 +/- 0.5 min in four experiments performed at 22 degrees C. In addition, we determined the t 1/2 of HD in D 2 O containing 0.17 M sodium chloride to be 24 +/ 1 min in three experiments performed at 22 degrees C. As further proof of the existence of HD dissolved into D 2 O, deutero-hexane was used to extract the D 2 O HD solution. The resulting deutero-hexane solution was studied by 1 H NMR and GC/MS. The results obtained match those received from a standard deutero-hexane HD solution. These results demonstrate that HD can be identified in D 2 O with proton NMR and that proton NMR data can be used to monitor the subsequent solvolysis of HD. PMID- 20021165 TI - From the new editor-in-chief. PMID- 20021166 TI - Ecotoxicity of ethylene glycol monomethyl ether and its acetate. AB - Ethylene glycol monomethyl ether (EGME) and ethylene glycol monomethyl ether acetate (EGMEA) have been tested for their acute and chronic toxicity to various organisms occupying different trophic levels in the aquatic ecosystems. The results obtained in this study and those collected from the literature clearly reveal that EGME does not present short- or long-term ecotoxic effects in the ranges of concentrations likely to be found in aquatic environments. Indeed, in general, concentrations of 1000 to 10,000 mg/L of EGME are necessary before significant adverse effects can be observed in aquatic species. Conversely, acute toxicity occurs in fish at about 50 mg/L of EGMEA, and reproduction of Ceriodaphnia dubia is affected by 0.06 mg/L of this chemical. A teratogenic effect-with a specific malformation of the eyes-occurs in Xenopus laevis in the presence of 75 mg/L of EGMEA. This study was partially supported by the French Ministry of the Environment as part of the PNETOX program (1998). PMID- 20021167 TI - Ecotoxicity of ethylene glycol monobutyl ether and its acetate. AB - Ethylene glycol monobutyl ether (EGBE) and ethylene glycol monobutyl ether acetate (EGBEA) were tested for their acute and chronic toxicity to various organisms occupying different trophic levels in the aquatic ecosystems. The obtained results and those collected from the literature clearly show that EGBE affects the survival, growth, and reproduction of aquatic organisms only at concentrations of approximately 100 mg/L or more. EGBEA appears to be slightly more ecotoxic to aquatic species. At 1000 and 10,000 mg/L, both chemicals strongly affect the early stages of development of the Japanese oyster, but such concentrations are too high to be found in aquatic environments. Micronucleus tests on Xenopus laevis show that EGBE and EGBEA are not genotoxic in the range of concentrations tested. Consequently, neither chemical presents a risk to the aquatic environment. EGBE does affect the gonadosomatic index of adult male zebra fish (Danio rerio); however, these are preliminary results and must be confirmed by additional experiments. This study was partially supported by the French Ministry of the Environment as part of the PNETOX program (1998). PMID- 20021168 TI - Cadmium uptake by radishes from soil contaminated with nickel-cadmium batteries: toxicity and safety considerations. AB - The objective of this investigation was twofold: (1) to determine the contamination of soil by cadmium (Cd) from commonly used nickel-cadmium (Ni-Cd) batteries utilized in electronic devices and (2) to determine the uptake of Cd in common garden radishes. Under normal controlled conditions, the study utilized depleted batteries; new, undamaged batteries; and new, damaged batteries. The data revealed that the contamination of the soil by Cd was significantly greater in the presence of new, damaged batteries than in the presence of the new, undamaged batteries or depleted batteries. In another set of experiments, the uptake of Cd was determined in radishes grown in the control soil as well as in the, Cd-contaminated soil. The findings revealed that Cd uptake was significantly greater by the leaf and stem grown in the soil contaminated with new, damaged batteries. These results indicate that improper disposal of Ni-Cd batteries contaminates the soil and leads to enhanced Cd levels in garden vegetables, which can pose a serious threat to human health. Presented in part at the Annual Meeting of the Semiconductors Safety Association, April 10, 2001, New Orleans, LA. The authors thank the Semiconductor Safety Association and the Dean, Dr. Betty Blodgett, for their encouragement and support; Dr. Harry Fannin for analyzing the data; and Mrs. Debra M. Britton and Mrs. Robin B. Doss for their assistance in the preparation of this manuscript. PMID- 20021169 TI - Spectrophotometric analysis of solubilized rat hair proteins following intraperitoneal injection of 2,5-hexanedione. AB - The neurotoxic industrial solvents n-hexane and methyl n-butyl ketone are toxic by virtue of their common metabolite, 2,5-hexanedione (2,5-HD). Our previous work showed that pyrrole-like substances in solubilized rat hair proteins from rats injected (ip) daily with 2,5-HD demonstrated maximal absorbance in the 530-nm spectral region following reaction with Ehrlich's reagent (p dimethylaminobenzaldehyde). Modification of the current analytical methods of achieving high specificity and lower detection limits with small sample quantities could have important implications for monitoring human populations. Adult male Sprague-Dawley rats were housed in individual metabolic cages with food and water provided ad libitum. Individual rats were injected (ip) daily with either 50 mg/kg 2,5-HD or physiologic-buffered saline (PBS). Plucked hair samples (dorsal, right flank, and left flank) were obtained from each rat before and at 7 day intervals after exposure to 2,5-HD or PBS for 28 days. Hair proteins solubilization and extraction procedures were adapted from earlier studies. We read 1 mL of dialyzed hair protein solution (2,5-HD or PBS control) against a reference cuvette containing water. Analyses utilized a Shimadzu UV 160 V recording spectrophotometer at an absorbency spectral range of 450 to 600 nm. In all spectral tracings, absorbance maxima (at 530 nm) characteristic of pyrrole like substances were detected only in samples from 2,5-HD-treated rats. Absorbance at 530 nm was detected starting at Day 7 after exposure. The authors acknowledge Dr. Richard Whorton and Dr. Barbara Buckley for advice and for the use of their spectrophotometric equipment and Dr. Lowell A. Goldsmith for his help in our choice of the subject studied. This work was supported by the Walter P. Inman Memorial Fund in an award from Duke University to Dr. Leon Lack. PMID- 20021170 TI - Application and detection of (14)c-hd in two mouse models. AB - The CD1-haired mouse and the SKH-hairless mouse are two animal models that have been used to evaluate sulfur mustard (HD) exposure and protection in our laboratory. In a recent study we observed that a substance P inhibitor protected the haired mouse ear against an HD solution, but the same drug was not successful in protecting the hairless mouse against HD vapor. This experiment prompted us to compare HD exposures between these models. We determined the (14)C content in the skin after exposures to HD containing (14)C-HD. Rate curves were generated for applications of (1) HD in methylene chloride to the haired mouse ear; (2) HD in methylene chloride to the hairless mouse dorsal skin; and (3) saturated HD vapor to the hairless mouse dorsal skin for 6 min. The curves showed a reduction in (14)C disintegrations per min in animals euthanized 0 to 2 h postexposure. The largest percentage of decrease of (14)C content in skin occurred within 30 min of HD challenge for all exposures. An 8-mm skin-punch biopsy and a 14-mm annular skin section surrounding the region of the 8-mm skin punch were taken from the hairless mouse dorsal skin exposed to HD in methylene chloride. The ratio of the (14)C content in the 8-mm skin punch to that in the surrounding 14-mm annular skin section was 7.3, demonstrating that the HD application spreads beyond the initially biopsied site. A concentration/time value of 6.3 mug/cm(2)/min was determined by counting skin (14)C disintegrations per minute in animals euthanized immediately after exposure to saturated HD vapor. Determinations of the amount of HD showed that similar quantities of HD, 0.4 mg, were detected on each model. These results contribute to a better quantitative understanding of HD application in the haired and hairless mouse models. PMID- 20021171 TI - A new method of estimating the cytotoxic effects of suspended particulate matter on cultured alveolar macrophages in vitro. AB - A new method has been devised for evaluating the cytotoxicity of suspended particulate matters in vitro. It uses the proliferative capacity of mouse alveolar macrophages, estimated by the uptake of (3)H-tymidine, as an index of cytotoxicity, and provides a simple and rapid means of evaluating the cytotoxicity of suspended particulate matter collected on a filter by a personal air sampler. A significant decrease in (3)H-thymidine uptake (an indication of impaired proliferation) could be detected in the range of approximately 0.31 to 1.03 mug of nickel sulfide particles per square centimeter of culture dish, suggesting the relatively great sensitivity of this method to particulate toxicants. A preliminary application of this method to suspended particulate matter collected in an urban atmosphere indicated that 3 hours of sampling, corresponding to that in about 0.72 cubic meters of air, were sufficient for evaluation of toxicity by this method. PMID- 20021172 TI - Preliminary study to test the feasibility of using a local lymph node type of approach to characterize respiratory allergens. AB - There is a critical need to develop animal models that can characterize the potential of respiratory allergy. Dust-mite allergens are one of the major etiological agents in the induction of allergy and asthma in humans. In this study, the effects of intratracheal injection with dust-mite allergen were investigated by analyzing the in vivo proliferative response of lung-draining hilar lymph nodes and histopathological changes in the lung parenchyma. Balb/c mice were inoculated intratracheally with dust-mite allergens, a mixture of Dermatophagoides farinae and D. pteronyssinus dissolved in phosphate-buffered saline, or with an equal volume of saline alone. After 1 week, all the mice were injected intravenously with radioactive (3)H-thymidine and sacrificed 5 h later so as to assess the radioactivity incorporated into the hilar lymph nodes. The results indicated a marked increase in the proliferative response in the hilar lymph nodes of the animals treated with the dust-mite allergen as compared to the response of the control group. Treatment with dust-mite allergen also caused perivascular and interstitial eosinophilic inflammation of the lungs, hyperplasia of bronchus-associated lymphoid tissue, and an increase in the eosinophil peroxidase activity in the lungs. These results indicate that intratracheal injection with dust-mite allergen can trigger a number of changes consistent with respiratory allergy, including an increased proliferation in the draining lymph nodes. PMID- 20021173 TI - Alteration in aromatic hydroxylation and lipid oxidation status in the lungs of rats exposed to ozone. AB - Fischer 344 rats were exposed to ozone by inhalation to identify sensitive indices of acute exposure. 5-Aminosalicylic acid (5-ASA) hydroxylation in bronchoalveolar lavage (BAL), an indicator of hydroxyl radical (*OH) formation, and lipid oxidation in various regions of airways, representing oxidative stress, were measured to verify whether they can function as markers of exposure. BAL cells and supernatants taken from rats that received saline or 5-ASA (ip, 50 mg/kg) prior to ozone exposure (0, 0.4, or 0.8 ppm for 4 h) were analyzed for products of lipid oxidation. *OH formation was assessed by analysis of the BAL supernatant for 5-aminotetrahydroxybenzoic acid (5-ATHBA), a hydroxylation product of 5-ASA. The tetrahydroxy derivative of 5-ASA was higher in the BAL of ozone-treated rats than in air controls, reaching significance (p <. 05) at 0.8 ppm of ozone, The products of lipid oxidation propanal and hexanal were higher in BAL cells taken from rats exposed to ozone, reaching significance (p <. 05) at a 0.8 ppm ozone level, compared to air control animals, irrespective of whether they received saline or 5-ASA prior to ozone exposure. Increases in cholesterol levels were also seen in BAL cells after rats were exposed to ozone. However, there were no significant dose-related changes in the lipid oxidation products in BAL supernatants after exposure to ozone. Lipid oxidation products in BAL cells and 5-ATHBA in lavage exhibited the potential to serve as markers of ozone exposure. This work was supported by Health Canada (#4320105) and Toxic Substances Research Initiatives (TSRI #60). PMID- 20021174 TI - Dose-dependent excretion of unconjugated 3-butene-1,2-diol measured in urine with a gc/ms after 1,3-butadiene exposure. AB - 1,3-Butadiene is clearly carcinogenic, and it has a complex pathway of metabolic transformation, as it forms several reactive intermediates. We have previously shown that butadiene diolepoxide is a key metabolite in DNA and hemoglobin adduct formation. Here we report the analysis of 3-buten-1,2-diol-the precursor of butadiene diolepoxide-in urine samples from rats exposed to butadiene by means of inhalation. Urine samples were extracted with isopropanol, and extracts were analyzed using a gas chromatograph and mass spectrometer. Selected ion monitoring was performed by using ion 57 m/z; a retention time allowed reliable analysis. The analysis showed a linear excretion of 3-buten-1,2-diol during the 5 days of 6 h exposures and 18-h recovery times between exposures. The daily correlation coefficient (r) values varied from 0.9945 to 0.9999. A 6-h exposure of rats to 1000 ppm 1,3-butadiene resulted in a mean urinary concentration of 3-buten-1,2 diol of approximately 38 mg/L. Urine samples were also collected during the recovery times, and the extracts were analyzed. The linearity of the excretion during the recovery times showed r values ranging from 0.6932 to 0.9813; 0.5 mg/L of 3-butene-1,2-diol was detected in urine samples excreted after exposure to 1000 ppm 1,3-butadiene. The data demonstrated that butadiene monoepoxide was converted into 3-buten-1,2-diol which, to some extent, was excreted in urine in a nonconjugated form. The excretion of 3-buten-1,2-diol was prompt, with about 98% of the compound being excreted during exposure. We acknowledge Mr. Yrjo Peltonen for his very skillful assistance in maintaining a stable BD atmosphere during animal exposure, and we thank Ms. Marja Pihlaja for the animal care. PMID- 20021175 TI - The state of polio vaccination in the world: the case for continuing routine vaccination. AB - This article reviews the literature concerning polio vaccination and uses that information to evaluate the World Health Organization's announced plans to end polio vaccination early in the 21st century. The safe elimination of polio vaccination would require the elimination of all polio cases, including paralytic cases, mild, nonparalytic cases, and vaccine-associated cases; proof that polio does not have any nonhuman alternative host; proof that no one alive can still shed the poliovirus; proof that no poliovirus persists in the environment; and proof that no live poliovirus, either the wild type or the vaccine strain, exists in medical or terrorist hands. Because it does not seem possible to ensure that ending vaccination against polio would be safe, and given the fact the polio is a terrible and untreatable disease, it would seem prudent to continue polio vaccination for the foreseeable future. PMID- 20021177 TI - Making a difference in publishing mechanistic toxicology research and innovative toxicological methods. PMID- 20021176 TI - A sensitive method of monitoring exposure to 3-bromobenzanthrone in industrial dyestuff workers. AB - 3-Bromobenzanthrone (3-BBA) is an anthraquinone dye intermediate widely used for the synthesis of a variety of dyes. The monitoring of 3-BBA exposure in dyestuff industry workers has not been possible until now as no procedure has been available. In this article, the fluorescence properties of 3-BBA has been utilized to develop a quantitative method for the detection of this dye intermediate. The procedure allows the measurement of trace quantities of 3-BBA in biological specimens, including urine, serum, liver, and feces, in experimental studies. The method involves extraction of biological samples with an equal volume of a chloroform-methanol mixture (1:1, v/v) followed by measurement of the relative fluorescence intensity at excitation maxima of 400 nm and emission maxima of 530 nm. The detection limit was found to be as low as 50 ng of 3-BBA. The procedure can be routinely used to screen for the presence of 3 BBA in biological fluids, especially urine, as a measure of exposure to 3-BBA in dyestuff workers. The authors are thankful to Dr. P. K. Seth, Director, Industrial Toxicology Research Centre, for his keen interest in this study. The secretarial assistance of Mr. K. G. Thomas is duly acknowledged. RPS is thankful to University Grant Commission, New Delhi, for the award of a Senior Research Fellowship. PMID- 20021178 TI - Idiosyncratic drug hepatotoxicity revisited: new insights from mechanistic toxicology. AB - Drug-induced hepatotoxicity is a serious problem and has repeatedly led to the withdrawal of a drug after successful launching. Although reactive metabolites (which are predictable) are often involved, genetic or acquired host factors (which are unpredictable) can increase the penetrance and expressivity of the potential hepatotoxicity in a small subset of patients. The molecular mechanisms underlying liver injury are largely unknown. Evidence suggests, however, that there are four major modes of action: covalent modification of target proteins and oxidoreductive stress; immune-mediated reactions; interference with hepatobiliary export; and mitochondrial injury. A better prediction will include new animal models, new biomarkers, and genomics/transcriptomics analysis. PMID- 20021179 TI - Rodent toxicity and nongenotoxic carcinogenesis: knowledge-based human risk assessment based on molecular mechanisms. AB - It is necessary to determine whether chemicals or drugs have the potential to pose a threat to human health. Chemicals that can damage DNA are detected in short-term assays, but the detection of nongenotoxic carcinogens relies upon bioassays in laboratory animals. However, there are marked differences between rodents and humans in response to nongenotoxic carcinogens, which makes the relevance of rodent data to human risk assessment questionable. Here, we address the background issues concerning rodent nongenotoxic carcinogenesis and then focus upon peroxisome proliferators, chloroform, and dioxins as examples of toxicants that cause rodent-specific oxidative stress, cell proliferation, and the suppression of apoptosis. In the case of peroxisome proliferators and dioxins, this response is receptor-mediated. The evidence presented suggests that, at least for some toxicants, the molecular mechanisms of the rodent carcinogenic responses do not operate in humans; this is discussed in the context of human risk assessment. Finally, consideration is given to incorporating mechanism-based information into risk assessment for regulatory purposes. PMID- 20021180 TI - Ornithine decarboxylase activity in tissues from rats exposed to 60 Hz magnetic fields, including harmonic and transient field characteristics. AB - Ornithine decarboxylase (ODC) activity is used widely as a biomarker for tumor promotion in animal model systems. Several previous studies have reported increases in ODC activity in tissues of rats exposed to 60 Hz magnetic fields. The goals of this study were to confirm these findings and to determine whether ODC activity is increased in tissues of animals exposed to magnetic fields containing complex metrics. Three experiments were conducted in male F344 rats. Each study included a sham control group and a group exposed to pure continuous 60 Hz fields (0.2 mT). Additional groups included animals exposed to randomly time-varying 60 Hz fields (range of 0.02 to 0.2 mT); intermittent 60 Hz fields (2 mT) with on-off cycles ranging from 5 s to 5 min; pure continuous 180 Hz fields (2 mT); 60 Hz fields with a superimposed 3rd harmonic (total field strength, 2 mT); 60 Hz fields with superimposed third, fifth, and seventh harmonics (total field strength, 2 mT); 60 Hz fields (2 mT) with superimposed transients; and randomly time-varying 60 Hz fields (range of 0.02 to 0.2 mT) with superimposed transients. After 4 weeks of exposure (18.5 h/day), eight animals per group were euthanized within 1 h of magnetic field deactivation. Homogenates of liver, kidneys, spleen, and brain were prepared from each animal, quick-frozen, and shipped for analysis by four independent laboratories. No consistent pattern of differences in the ODC activity among experimental groups was found either within a laboratory or among laboratories. The results do not support the hypothesis that exposure to extremely low frequency magnetic fields stimulates ODC activity. PMID- 20021181 TI - Validation of MRI measurement of cardiac output in the dog: the effects of dobutamine and minoxidil. AB - The use of magnetic resonance imaging (MRI) for the measurement of cardiac output parameters in anesthetized adult male beagle dogs has been validated against a widely accepted thermodilution method. Using a multislice cine gradient echo MRI method to acquire images of the entire heart, left ventricular lumen volumes were measured at systole and diastole in seven animals. Cardiac output correlated well (R 2 = 0.88) with thermodilution measurements made in a parallel manner, both before and during acute stimulation with the inotrope dobutamine. In a chronic study of changes in cardiac morphology and function brought about by the antihypertensive minoxidil, MRI reliably detected the expected increases in stroke volume (28%) and cardiac output (58%) resulting from neural reaction to decreased blood pressure. Left ventricular lumen enlarged as well in response to fluid retention and plasma volume increase. Two in four minoxidil-treated animals also developed clear MRI-visible pericardial effusion. PMID- 20021182 TI - Variance of Microsomal Protein and Cytochrome P450 2E1 and 3A Forms in Adult Human Liver. AB - Differences in the pharmacokinetics of xenobiotics among humans makes them differentially susceptible to risk. Differences in enzyme content can mediate pharmacokinetic differences. Microsomal protein is often isolated from liver to characterize enzyme content and activity, but no measures exist to extrapolate these data to the intact liver. Measures were developed from up to 60 samples of adult human liver to characterize the content of microsomal protein and cytochrome P450 (CYP) enzymes. Statistical evaluations are necessary to estimate values far from the mean value. Adult human liver contains 52.9 +/- 1.476 mg microsomal protein per g; 2587 +/- 1.84 pmoles CYP2E1 per g; and 5237 +/- 2.214 pmols CYP3A per g (geometric mean +/- geometric standard deviation). These values are useful for identifying and testing susceptibility as a function of enzyme content when used to extrapolate in vitro rates of chemical metabolism for input to physiologically based pharmacokinetic models which can then be exercised to quantify the effect of variance in enzyme expression on risk-relevant pharmacokinetic outcomes. PMID- 20021183 TI - Incorporation of protein-binding kinetics and carrier-mediated membrane transport into a model of chemical kinetics in the isolated perfused rat liver. AB - Nonlinear processes present difficult problems in extrapolating toxicological kinetics from one dose or species to another. A model that includes the details of these processes will aid in identifying those conditions under which a linear extrapolation is not valid. Previously, a kinetic model of the perfused rat liver system was presented that included the effects of equilibrium protein binding, diffusion-based and saturable membrane transport, metabolism, and biliary excretion. This model has been extended here to account for two complex processes. First, the kinetics of chemical association and dissociation from binding sites in various compartments are explicitly included to allow for the possibility that the binding of the compound is not in equilibrium. Second, mediated transport via a simple four-state carrier in the membrane has been included at both the sinusoidal and biliary membranes of the liver cell. This enables inclusion of carrier-specific transport processes (such as selective transport against a concentration gradient) in modeling the kinetics in perfused rat liver experiments. Simulations demonstrating the effects of each of these processes on observable state variables have been conducted. Physiological conditions that elicit nonlinear behavior have been identified. PMID- 20021184 TI - Oxidant-antioxidant status in oral precancer and oral cancer patients. AB - The present investigation was undertaken to examine the blood levels of lipid peroxides and the antioxidants superoxide dismutase, catalase, reduced glutathione, glutathione peroxidase, and glutathione-S-transferase in oral precancer, preoperative, postoperative, and recurrent oral cancer patients with age- and sex-matched normal healthy subjects as controls. In patients with oral precancer and oral cancer, enhanced lipid peroxidation was accompanied by antioxidant depletion. These changes were more pronounced in patients with recurrent oral tumors. Twelve weeks after surgery, decreased lipid peroxidation was accompanied by increased antioxidant levels. The results of this study indicate that imbalance in the redox status of oral cancer patients may be due to enhanced lipid peroxidation and compromised antioxidant defenses. PMID- 20021185 TI - Chemoprotection against N-methyl-n'-nitro-N-nitrosoguanidine-induced oxidative stress by s-allylcysteine, a garlic constituent, in wistar rats. AB - S-allylcysteine (SAC), a water-soluble garlic constituent, was tested for its protective effect against N-methyl-N'-nitro-N-nitrosoguanidine (MNNG)-induced oxidative stress in male Wistar rats. Three different concentrations of SAC (100, 200, and 400 mg/kg (-) 1 body weight) were administered intragastrically for 5 consecutive days, followed by intragastric intubation of MNNG 1(1/2) h after the final administration. Measurement of lipid peroxidation, reduced glutathione (GSH), glutathione peroxidase (GPx), and glutathione S-transferase (GST) in the stomach, liver, and erythrocytes was used to monitor the chemoprotective effects of SAC. Enhanced lipid peroxidation in MNNG-treated rats was accompanied by a significant decrease in glutathione and in the activities of GPx and GST. Administration of SAC at the concentration of 200 and 400 mg/kg (-) 1 body weight significantly reduced the concentration of lipid peroxides and enhanced the levels of GSH-dependent antioxidants and detoxifying enzymes. Our findings suggest that SAC exerts chemoprotective effects, even at a concentration of 200 mg/kg (-) 1 body weight, against MNNG-induced oxidative stress by decreasing lipid peroxidation and enhancing the antioxidant status. PMID- 20021186 TI - Effect of chromium on certain aspects of metabolic toxicities. AB - The impact of chromium exposure was studied in the liver, kidney, testis, spleen, cerebrum, and cerebellum of male Wistar rats (80-100 g body weight). It was observed that treatment of rats with chromium (ip, at a dose of 0.8 mg/100 g body weight/day) for a period of 28 days caused significant increase in chromium content while lowering the body weight along with the organ weight, except for the liver. It was also observed that there was a significant decrease in the DNA content of various organs tested. Also, a significant decrease in RNA content was observed in all the organs tested except for the testes. The liver, cerebrum, and cerebellum showed significant decreases in total protein content in chromium treated animals, whereas the kidney, testes, and spleen showed insignificant alterations. The RNase activity was found to be significantly increased only in the testes and cerebrum. Pronase activity was significantly increased in the tissues, except for the liver. The glutamic-pyruvic acid transaminase activity decreased in all the tissues studied. On the other hand, glutamic-oxaloacetic acid transaminase activity increased in the liver, cerebrum, and cerebellum while decreasing in the kidney and spleen. It is suggested that chromium exposure at the present dose and duration induces metabolic toxicity in the form of depressive effects on nucleic acids and altered activities of RNase, pronase, and transaminases in tissues. The extent of such alterations varies from tissue to tissue and is in some cases diverse in nature. PMID- 20021187 TI - Determination of Energy Charge Potential in the C6 Glioma and the HepG-2 Cell Culture. AB - The energy charge potential (ECP) is the ratio among adenosine 5'-triphosphate (ATP), -diphosphate (ADP), and -monophosphate (AMP); it reflects the high-energy bond content in biological systems. Under normal physiological conditions, the value of ECP is between 0.7 and 1.0. In this study, two different methods were used to extract adenosine nucleotides (ATP, ADP, and AMP) for determination of the ECP in the C6 glioma and the HepG-2 human hepatoma cell lines. In the first method, cells were removed from culture plates by trypsinization before the nucleotides were extracted using perchloric acid. Some of the trypsinized cells were used for a cell count, thereby allowing the calculation of an average nucleotide level per cell. In the second method, perchloric acid was added directly to the culture plate immediately after the medium was removed. This method resulted in an instant termination of any enzymatic process that might degrade the nucleotides and alter their levels in cells. Adenosine nucleotides were determined by high-performance liquid chromatography. The direct extraction method yielded a total adenosine nucleotide (TAN) level more than twofold higher than that obtained following trypsinization. For the C6 glioma cells, the ratio of ATP to ADP to AMP in the TAN pool was 0.70:0.15:0.15, as compared to 0.36:0.27:0.37 when cells were extracted following trypsinization. Similarly, for the HepG-2 cells, the ratio was 0.62:0.15:0.23 following direct extraction, as compared to 0.15:0.28:0.57 following trypsinization. Using the results from the direct extraction method, the value of the cellular ECP was 0.78 +/- 0.06 for the C6 glioma cells and 0.70 +/- 0.06 for the HepG-2 cells. Compared to the ECP values obtained using the trypsinization method (0.50 +/- 0.05 and 0.29 +/- 0.12, respectively), the ECP values derived using the direct extraction method were significantly greater in both cell lines and were suggestive of the energy status of normal, healthy cells. PMID- 20021188 TI - A rapid in vitro method of identifying contact allergens and irritants. AB - A rapid in vitro method has been developed to screen skin allergens and irritants. It has been shown by others that allergens induce the secretion of interleukin (IL)-1 betaby dendritic cells and macrophages, which are known antigen-presenting cells present in blood, and that irritants induce the secretion of IL-1 alphaby the same cells. We have shown that the allergens nickel, chromium, isoeugenol, and dinitrofluoro benzene induce the secretion of IL-1 betaat levels that are two- to threefold higher than those of controls and that the nonallergens and irritants sodium dodecyl sulfate, Tween-20, acetic acid, sodium hydroxide, and dimethyl sulfoxide fail to induce such a response. All the chemical allergens tested induced the secretion of IL-1 alphaat concentrations two- to fivefold higher than those of controls, depending on the concentration and the particular irritant. Allergen-induced T-cell proliferation has also been demonstrated by the expression of IL-2 as well as 3-(4,5 dimethylthiazol-2-yl)-2,5-diphenyltetrazolim bromide reduction (MTT) assay (10 50% more than control). All the allergens and irritants caused cytotoxicity at higher doses, as indicated by the release of lactate dehydrogenase. This method can be used to differentiate the allergens from the irritants in a simple and convenient manner. PMID- 20021189 TI - Hepatocyte culture as an in vitro model for evaluating the hepatotoxicity of food borne toxicants and microbial pathogens: a review. AB - Hepatocyte culture is a well-established, well-characterized, and widely used in vitro tool for pharamacological and toxicological studies. The hepatocytes of a wide range of species, including humans, can be maintained in culture with a high metabolizing capacity for several days under closely controlled and easily manipulated conditions. Numerous studies have reported good correlation between in vitro hepatocytes and in vivo siutations. They have been widely used for studies of the metabolism and of the toxicity and mechanisms of action of chemicals and drugs, for the screening of mutagens, carcinogens, and micotoxins, for virulence assessment of microbial pathogens and viruses, for qualitative and quantitative interspecies comparison, and for genomics and proteiomics studies. Hepatocytes, especially human hepatocytes, are used for preclinical drug evaluation and screening as well as for studies of drug metabolism, toxicity, interactions, and structure-activity relationships. They can be used for evaluating the hepatotoxicity of herbal products, dietary supplements, food additives, food-borne toxicants, and microbial pathogens. The results obtained from such in vitro screenings can be used for in vivo studies to asses the safety of test materials of interest. At the present time, there is insufficient evidence for their use in quantitative risk assessment. However, they are suitable for use in qualitative hazard assessment, which may be used for quantitative risk analysis. PMID- 20021190 TI - HPLC/DAD Screening Method for Selected Psychotropic Drugs in Blood. AB - The influence of experimental conditions on gradient high-performance liquid chromatography/diode array detector separation and identification of 13 psychotropic drugs belonging to two groups--phenothiazines and tricyclic antidepressants--was examined. The main interaction effects of three experimental factors were determined according to a 2 3 factorial design, and the optimum conditions of analysis were searched for. The degree to which the chromatographic peaks overlap is taken into account in a proposed criterion for separation quality. The screening analysis proposed was tested on whole blood samples spiked with mixtures of the examined drugs. The method was characterized by such validation parameters as relative retention times, absorbance ratios at two wavelengths, detection limits, linearity ranges, and extraction recoveries. The method proved to be a suitable tool for the identification of psychotropic drugs tested for forensic purposes. PMID- 20021191 TI - A simple and highly sensitive spectrophotometric method for the determination of cyanide in equine blood. AB - An epidemiological association among black cherry trees (Prunus serotina), eastern tent caterpillars (Malacosoma americana), and the spring 2001 episode of mare reproductive loss syndrome in central Kentucky focused attention on the potential role of environmental cyanogens in the causes of this syndrome. To evaluate the role of cyanide (CN (-)) in this syndrome, a simple, rapid, and highly sensitive method for determination of low parts per billion concentrations of CN (-) in equine blood and other biological fluids was developed. The analytical method is an adaptation of methods commonly in use and involves the evolution and trapping of gaseous hydrogen cyanide followed by spectrophotometric determination by autoanalyzer. The limit of quantitation of this method is 2 ng/mL in equine blood, and the standard curve shows a linear relationship between CN (-) concentration and absorbance (r >. 99). The method throughput is high, up to 100 samples per day. Normal blood CN (-) concentrations in horses at pasture in Kentucky in October 2001 ranged from 3-18 ng/mL, whereas hay-fed horses showed blood CN (-) levels of 2-7 ng/mL in January 2002. Blood samples from a small number of cattle at pasture showed broadly similar blood CN (-) concentrations. Intravenous administration of sodium cyanide and oral administration of mandelonitrile and amygdalin yielded readily detectable increases in blood CN (-) concentrations. This method is sufficiently sensitive and specific to allow the determination of normal blood CN (-) levels in horses, as well as the seasonal and pasture-dependent variations. The method should also be suitable for investigation of the toxicokinetics and disposition of subacutely toxic doses of CN (-) and its precursor cyanogens in the horse as well as in other species. PMID- 20021192 TI - Associations of Blood Lead Levels, Kidney Function, and Blood Pressure with delta Aminolevulinic Acid Dehydratase and Vitamin D Receptor Gene Polymorphisms. AB - The association of lead toxicity with delta -aminolevulinic acid dehydratase (ALAD) and vitamin D receptor gene polymorphisms was examined in 216 workers occupationally exposed to lead. Urinary N-acetyl- beta-glucosaminidase (UNAG) activity and albumin concentration, adjusted by creatinine, were measured as biomarkers of nephrotoxicity. Genotyping was performed using polymerase chain reaction and restricted fragment length polymorphisms. Blood pressure and blood and urinary lead concentrations were measured. No statistically significant difference in blood lead concentrations was observed with the ALAD genotype, although the geometric mean of blood lead concentrations was higher in battery workers with the ALAD-12 genotype (50.95 mug/dL) than in workers with the ALAD-11 genotype (42.95 mug/dL). Battery workers with the vitamin D receptor (VDR)-B allele had higher mean blood lead concentrations (60.54 mug/dL) than did the workers with the VDR-bb genotype (51.65 mug/dL) (p <. 05) . Based on multiple linear regression analysis, the VDR genotype was associated with a blood lead level (p <. 021). The ALAD genotype was associated with a urinary lead level (p <. 009). Workers with the ALAD-12 genotype who also had blood lead concentrations equal to or greater than 40 mug/dL had higher UNAG activity (4.17 U/mmol.Cr) than the workers with the ALAD-11 genotype (1.27 U/mmol.Cr) (p =. 06). Compared with workers with the VDR-bb genotype, workers with the VDR-B allele had higher systolic blood pressure when the blood lead level was equal to or greater than 40 mug/dL (133.06 +/- 19.19 mmHg versus 123.12 +/- 15.69 mmHg, p =. 01). Lead workers with ALAD-12 had larger elevations in UNAG activity and increasing blood lead concentrations compared to lead workers with the ALAD-11 genotype. Lead workers with the VDR-B allele had greater elevations in systolic blood pressure and higher blood lead levels than did workers with the VDR-bb genotype. In summary, the ALAD-2 and VDR-B alleles may modify lead absorption and toxicity. PMID- 20021193 TI - Evaluation of the No Observed Adverse Effect Level of Solvent Dimethyl Sulfoxide in Drosophila melanogaster. AB - Dimethyl sulfoxide, a solvent commonly used in toxicological studies, was investigated for its cytotoxic potential and its effect on development and reproductive performance in transgenic Drosophila melanogaster (hsp70-lacZ) Bg 9. Various concentrations (0.0, 0.1, 0.3, 0.5, 1.0, 2.0, and 3.0%) of the solvent were mixed with food and fed to the flies and larvae. The toxic effects were studied by examining hatchability, emergence, fecundity, reproductive performance, and hsp70 expression by means of in situ beta-galactosidase staining in the tissues of third-instar larvae and in the reproductive organs of male and female flies of the strain. Our results showed that dimethyl sulfoxide at and above 0.5% of dietary concentration evoked cytotoxicity, as evidenced by hsp70 expression in the larval tissues, and also impaired the development and reproductive performance of the flies. This study suggests that the no observed adverse effect level of dimethyl sulfoxide is 0.3% of dietary concentration for developmental toxicity, reproductive toxicity, and cytotoxicity studies in Drosophila melanogaster. PMID- 20021194 TI - Comparative Measurement of In Vitro T-2 Toxin Cytotoxicity using Three Different Cytotoxicity Assays. AB - T-2 toxin is a secondary fungal metabolite produced by various species of fusarium. In this study, the in vitro cytotoxicity of T-2 toxin in the human hepatoma cell line (HepG2) was determined using three methods, including neutral red dye adsorption, trypan blue dye exclusion, and uptake of radiolabeled 3 H thymidine. The 50% lethal concentration of T-2 toxin was found to be similar for all three, accounting for 2.39, 2.96, and 3.44 ng/mL, respectively, as determined by these assays. Significant correlations were also observed among the three methods (p <. 05 through p <. 005). These results suggest that all three methods are reliable and are comparable for determining the 50% lethal concentration of T 2 toxin. It is concluded that the neutral red cytotoxicity assay may be preferable to other methods because of its simplicity, its use of nonradioactive materials, and its objectivity. PMID- 20021195 TI - Erratum. PMID- 20021196 TI - Serial phenotypic analysis of mouse peripheral blood leukocytes. AB - Repeated phenotypic analysis of mouse peripheral blood leukocytes over short periods of time (2 weeks) has been difficult because of the very limited volumes of blood available under guidelines of the Institutional Animal Care and Use Committee. The loss of leukocytes and variations among laboratories during conventional flow cytometry sample preparation based on lysing and repeated washing have been limiting factors when measuring multiple parameters in small samples. We describe a method of phenotypic analysis using a no-lyse, no-wash staining technique combined with fluorescent triggering for data collection that can be performed on volumes of 20 muL or less of whole blood per set of markers in one tube. This method allows repeated phenotypic analysis of peripheral whole blood from mice. Fluorescent triggering with anti-CD45-PE/Cy5 antibody allows high-quality phenotypic data to be collected for CD4, CD8, TcR- beta, CD45R (B220), CD11b, and Gr-1 epitopes on leukocytes from mouse peripheral blood without lysis. The markers selected cover the major populations in peripheral mouse blood. Reproducibility and time-course data are presented for sampling periods as long as 4 weeks. Data produced by flow cytometers manufactured by two different companies show well-correlated results. An instrument equipped with a gated amplifier or a photomultiplier tube suitable for Cy7 conjugates could measure additional parameters. Because of interference from unlysed erythrocytes, scatter parameters are not useful for identifying cell populations with this method. PMID- 20021197 TI - Cold and cryopreservation of monkey liver slices. AB - Both cynomolgus and rhesus monkeys are utilized in chemical toxicity screening and drug development studies by industry and government laboratories. Along with better utilization of their tissue for in vivo studies, it would be advantageous if the tissue could be cold-preserved or cryopreserved for future in vitro experimentation. Therefore, livers were excised from control monkeys and precision-cut tissue slices were prepared. The objective of this study was twofold: to compare cold-preservation solutions (V-7 and Viaspan) and to compare controlled-rate and vitrification cryopreservation protocols. Monkey liver slices were cold-stored in V-7 or Viaspan preservation solutions for 7 days. V-7 maintained slice viability for 5 days whereas Viaspan maintained slice viability for 1 day. In the controlled-rate freezing procedure, slices were exposed to 10% dimethyl sulfoxide and 90% fetal calf serum (FCS); cooled at the rate of 0.5 degrees C, 1 degrees C, or 12 degrees C per min to -70 degrees C; then placed into liquid nitrogen. Vitrification was accomplished by exposing slices stepwise to increasing concentrations of 1,2-propanediol (1.2, 2.4, and 4 M) in FCS with direct submersion into liquid nitrogen. In both protocols, slices were rewarmed quickly to 37 degrees C and then incubated in FCS for 4 h. Three viability parameters were used to measure slice viability - retention of potassium, leakage of lactate dehydrogenase, and synthesis of protein. Liver slices cryopreserved at a rate of 0.5 degrees C per min and vitrified successfully retained 80 to 90% of their viability. These results confirm the feasibility of functional cold preservation and cryopreservation protocols for monkey liver slices that would allow for a more efficient use of monkey tissue. PMID- 20021198 TI - The microbial biogenesis of aquatic biotoxins. AB - The biogenesis of small molecular nonprotein poisons has long been a mystery in the world of biotoxicology. The phylogenetic distribution, molecular structure, evidence of biogenesis, and possible mechanisms involving some of these poisons, such as saxitoxin, tetrodotoxin, ciguatoxin, palytoxin, maitotoxin, and brevitoxin, are briefly discussed. PMID- 20021199 TI - Detection by rapd of genetic alterations in vitro: amplification and conservation conditions of DNA extracts. AB - The technique of random amplified polymorphic DNA (RAPD) permits the study of genetic ecotoxicology without having to face the problem posed by the high number of chromosomes generally presented by the fish species used in traditional methods. Additionally, the use of in vitro systems allows the study of a large number of samples in environmental-risk assessment. Good standardization of the parameters involved in the RAPD reaction, such as primer concentration and the DNA template used and its integrity, is fundamental for obtaining reliable and repeatable results. This is especially important when the differences in DNA fingerprints between control cells and cells exposed to genotoxic agents are interpreted as toxic-dependent alterations. The use of more than one primer increases sensitivity in the detection of such differences, provided the amplification is carried out under optimum conditions. This article demonstrates that the conditions established for certain primers in previous studies can be acceptable for others, independent of the complexity of band patterns generated. Furthermore, the integrity of the DNA is shown to be stable for several months in the genomic extracts stored at 4 degrees C, which to a large extent facilitates the application of this methodology. PMID- 20021201 TI - Update on the potential of computer-aided diagnosis for breast cancer. PMID- 20021200 TI - Molecular cloning and enzymatic characterization of sheep CYP2J. AB - Cytochrome P450 (CYP) 2Js have been studied in various mammals, but not in sheep, as an animal model used to test veterinary drug metabolism. Sheep CYP2J was cloned from liver messenger RNA (mRNA) by RACE. The cDNA, after modification at its N- and C-terminals, was expressed in Escherichia coli and the sheep CYP2J protein, purified by chromatography, was 80% homologous to human and monkey CYP2J2. Reverse transcriptase-polymerase chain reaction (RT-PCR) experiments showed that CYP2J mRNA was expressed in liver, cortex, respiratory and olfactory mucosa, heart, bronchi, lung, spleen, small intestine and kidney. The purified enzyme was catalytically active towards aminopyrine, all-trans-retinoic acid, and particularly arachidonic acid forming 20-HETE, 19-HETE, and 18-HETE (about 86% of the total) and 14,15-, 11,12-, 8,9-, and 5,6-EETs (cis-epoxyeicosatrienoic acids; about 14% of total), with a regioselectivity similar to that shown by the mammalian CYP2J2s. PMID- 20021202 TI - Assessing parental contributions to childhood cancer risk. PMID- 20021203 TI - Neurosurgeons achieve world's first delivery of intra-arterial bevacizumab directly into brain tumor of glioblastoma multiforme patients. PMID- 20021204 TI - Tumor metabolism to blood flow ratio in pancreatic cancer: helpful in patient stratification? AB - Evaluation of: Komar G, Kauhanen S, Liukko K et al.: Decreased blood flow with increased metabolic activity: a novel sign of pancreatic tumor aggressiveness. Clin. Cancer Res. 15(17), 5511-5517 (2009). Novel methods in assessing pancreas cancer vascularization and metabolism are urgently needed for pretherapeutic patient stratification. Recently, noninvasive quantification of blood flow and metabolic activity of pancreatic tumors using [(15)O]H(2)O and [(18)F] fluorodeoxyglucose positron emission tomography/computed tomography imaging has been demonstrated to be a promising approach to assess these parameters. This also implied that high tumor metabolism and low blood flow (and thus probably also low tissue oxygenation) are linked with an aggressive cancer phenotype, and potentially reflect treatment resistance. Furthermore, such functional imaging might prove clinically that pancreatic cancer differs from other gastrointestinal cancer in its desmoplastic reaction and hypovascularity, which might well explain the enormous heterogeneity in treatment sensitivity. Therefore, determination of the metabolism to blood flow ratio could help in the decision on which (additional) chemotherapies or targeted agents to administer to a particular patient. Chemoresistance in pancreatic cancer might thus be regarded, at least in part, as a result of low drug delivery to the tumor cells, which is a consequence of a hypoxic environment contributing to the stromal reaction, rather than epithelial tumor cell resistance itself. PMID- 20021205 TI - miR-31: a master regulator of metastasis? AB - Evaluation of: Valastyan S, Reinhardt F, Benaich N et al.: A pleiotropically acting microRNA, miR-31, inhibits breast cancer metastasis. Cell 137(6), 1032 1046 (2009). Several microRNAs (miRNAs) regulate various steps of the metastatic cascade; however, they typically regulate both metastasis-related genes and genes involved in proliferation, cell-cycle regulation or apoptosis. miR-31 was determined to regulate a number of metastasis-related genes in breast cancer cells and tissues. Using a variety of approaches, it was demonstrated that cellular levels of miR-31 correlated with the cell's ability to invade and metastasize; cells with increased levels of the miRNA were less metastatic. These studies were extended to patient's tissues, demonstrating that breast cancer patients with higher miR-31 expression, or lower expression of the miR-31 target genes, had prolonged survival. A miRNA that works pleiotropically to regulate invasion and metastasis has been identified, suggesting a new pathway for therapeutic intervention of metastasis in breast cancer. PMID- 20021206 TI - Evaluation of a hepatitis B vaccination program in Taiwan: impact on hepatocellular carcinoma development. AB - Evaluation of: Chang MH, You SL, Chen CJ et al.; Taiwan study group: Decreased incidence of hepatocellular carcinoma in hepatitis B vaccinees: a 20 years follow up study. J. Natl Cancer Inst. 101, 1348-1355 (2009). This population-based study aimed to investigate whether a universal Taiwanese program, launched in 1984,has been able to prevent hepatocellular carcinoma (HCC) beyond childhood.The Incidence of HCC in Taiwan from 1983 to 2004 was assessed through two national cancer registries where age and sex-specific incidence were compared among a vaccinated and unvaccinated birth cohort with regression models. This study further characterized the 64 vaccinated cases that developed HCC in order to discriminate specific risk factors. PMID- 20021207 TI - Histologic tumor type and the rate of complete response after neoadjuvant therapy for esophageal cancer. AB - A review of the literature demonstrated that clinical evaluation cannot be used to determine 'complete response'. The different classification systems of the histopathologic response grading after neoadjuvant radiochemotherapy of esophageal carcinoma are summarized in this report. A systematic review of studies analyzing preoperative chemoradiation of squamous cell carcinoma (SCC) or adenocarcinoma (AC) of the esophagus demonstrated no significant difference in pathologic complete response (pCR) rates between the AC and SCC studies. Analyzing only the applied dose of radiation demonstrated that patients with AC required a higher dose than patients with SCC to achieve complete response. Incorporating chemotherapy administration does not markedly change the difference in required radiation dose. However, when the tumor does respond, the rate of pCR with increasing dosage of chemoradiotherapy increases more rapidly in AC patients than in SCC patients. PMID- 20021208 TI - Potential of ErbB4 antibodies for cancer therapy. AB - Antibodies targeting the extracellular domains of ErbB receptors have been extensively studied for cancer drug development. This work has led to clinical approval of monoclonal antibodies against the well-known oncogenes EGFR and ErbB2. Here we discuss the biological activities of ErbB4, a less-studied member of the EGFR/ErbB growth factor receptor family and speculate on the potential clinical relevance of antibodies targeting ErbB4. In addition to their significance as therapeutics, the role of ErbB4 antibodies in prognostic and predictive applications is surveyed. PMID- 20021209 TI - Emerging therapeutic targets in breast cancer bone metastasis. AB - In the past decade, our understanding of the molecular mechanisms that underlie breast cancer pathology and progression has dramatically improved. Using this knowledge, we have identified additional targets and developed novel therapeutic interventions in breast cancer. Together, these translational research efforts are helping to usher us into an age of personalized cancer therapy. Metastasis to bone is a common and devastating consequence of breast cancer. Bisphosphonates, which represent the current gold standard in bone metastasis therapies, are being improved with newer and more efficacious generations of these compounds being developed. Breast cancer growth in the bone requires activation of various signaling pathways in both cancer cells and stromal cells, including those that are stimulated by TGF-beta and RANKL, and mediated through the Src tyrosine kinase. Bone cells and cancer cells alike express promising targets for therapeutic intervention, including Cathepsin K, CXCR4 and GPNMB. In this article we discuss the molecular mechanisms behind these pro-metastatic molecules and review the most recent findings in the clinical development of their associated targeted therapies. PMID- 20021211 TI - Immunohistochemistry in the diagnosis of cutaneous neoplasms. AB - In recent years, there has been a significant increase in the number of diagnostic immunohistochemical stains available to pathologists and dermatopathologists. Immunohistochemistry has become an indispensable tool in dermatopathology, not only in diagnosis but also in the treatment and prognostication of cutaneous neoplasms. In this review we attempt to outline current, as well as new, immunohistochemical stains of relevance in the diagnosis and classification of cutaneous neoplasms. PMID- 20021210 TI - Depurinating estrogen-DNA adducts in the etiology and prevention of breast and other human cancers. AB - Experiments on estrogen metabolism, formation of DNA adducts, mutagenicity, cell transformation and carcinogenicity have led to and supported the hypothesis that the reaction of specific estrogen metabolites, mostly the electrophilic catechol estrogen-3,4-quinones, with DNA can generate the critical mutations to initiate breast and other human cancers. Analysis of depurinating estrogen-DNA adducts in urine demonstrates that women at high risk of, or with breast cancer, have high levels of the adducts, indicating a critical role for adduct formation in breast cancer initiation. Men with prostate cancer or non-Hodgkin lymphoma also have high levels of estrogen-DNA adducts. This knowledge of the first step in cancer initiation suggests the use of specific antioxidants that can block formation of the adducts by chemical and biochemical mechanisms. Two antioxidants, N acetylcysteine and resveratrol, are prime candidates to prevent breast and other human cancers because in various M in vitro and in vivo experiments, they reduce the formation of estrogen-DNA adducts. PMID- 20021212 TI - CXCL8 and its cognate receptors in melanoma progression and metastasis. AB - The incidence of melanoma is rising at an alarming rate and we are still awaiting an effective treatment for this malignancy. In its early stage, melanoma can be cured by surgical removal, but once metastasis has occurred there is no effective treatment. Recent findings have suggested multiple functional implications of CXCL8 and its cognate receptors, CXCR1 and CXCR2, in melanoma pathogenesis, thus underscoring their importance as targets for cancer therapy. This review provides an update on the roles of CXCL8 and its receptors in melanoma progression and metastasis. PMID- 20021213 TI - Future of convection-enhanced delivery in the treatment of brain tumors. AB - Gliomas are one of the most lethal forms of cancer. The poor prognosis associated with these malignant primary brain tumors treated with surgery, radiotherapy and chemotherapy has led researchers to develop new strategies for cure. Interstitial drug delivery has been the most appealing method for the treatment of primary brain tumors because it provides the most direct method of overcoming the barriers to tumor drug delivery. By administering therapeutic agents directly to the brain interstitium and, more specifically, to tumor-infiltrated parenchyma, one can overcome the elevated interstitial pressure produced by brain tumors. Convection-enhanced delivery (CED) has emerged as a leading investigational delivery technique for the treatment of brain tumors. Clinical trials utilizing these methods have been completed, with mixed results, and several more are being initiated. However, the potential efficacy of these drugs may be limited by ineffective tissue distribution. The development of computer models/algorithms to predict drug distribution, new catheter designs, and utilization of tracer models and nanocarriers have all laid the groundwork for the advancement of CED. In this review, we summarize the recent past of the clinical trials utilizing CED and discuss emerging technologies that will shape future CED trials. PMID- 20021215 TI - Role of systemic inflammatory response in predicting survival in patients with primary operable cancer. AB - Disease progression in cancer is dependent on the complex interaction between the tumor and the host inflammatory response. There is substantial evidence in advanced cancer that host factors, such as weight loss, poor performance status and the host systemic inflammatory response, are linked, and the latter is an important tumor-stage-independent predictor of outcome. Indeed, the systemic inflammatory response, as evidenced by an elevated level of C-reactive protein, is now included in the definition of cancer cachexia. This review examines the role of the systemic inflammatory response in predicting survival in patients with primary operable cancer. Approximately 80 studies have evaluated the role of the systemic inflammatory response using biochemical or hematological markers, such as elevated C-reactive protein levels, hypoalbuminemia or increased white cell, neutrophil and platelet counts. Combinations of such factors have been used to derive simple inflammation-based prognostic scores, such as the Glasgow Prognostic Score, the neutrophil:lymphocyte ratio and the platelet:lymphocyte ratio. This review demonstrates that there is now good evidence that preoperative measures of the systemic inflammatory response predict cancer survival, independent of tumor stage, in primary operable cancer. The evidence is particularly robust in colorectal (including liver metastases), gastro-esophageal and renal cancers. As described in this article, measurement of the systemic inflammatory response is simple, reliable and can be clinically incorporated into current staging algorithms. This will provide the clinician with a better prediction of outcome, and therefore better treatment allocation in patients with primary operable cancer. Furthermore, systemic inflammation-based markers and prognostic scores not only identify patients at risk, but also provide well defined therapeutic targets for future clinical trials. PMID- 20021216 TI - Ephrin expression and function in cancer. AB - Ephrins are cell membrane-associated signaling proteins bound by transmembrane Eph receptors on juxtaposed cells. Eph-ephrin interactions result in bidirectional signaling within both receptor- and ligand-bearing cells, with diverse consequences for cell morphology and behavior. Such interactions are especially important during early vertebrate development, and growing evidence has revealed equally important roles in adult-tissue homeostasis. As for the Eph receptors, abnormal expression of ephrins is associated with disease, especially cancer. The ephrins have received less attention than the Ephs in the literature, owing, in part, to their later discovery and that they are fewer in number. Here, we attempt to redress this imbalance and provide an 'ephrin-centric' discussion of the expression and function of ephrins in cancer. PMID- 20021217 TI - Clinical utility of circulating tumor cell measurement in the diagnosis of indeterminate lesions of the pancreas. AB - The quantification of circulating tumor cells has been historically problematic due to the different methods applied to their measurement. Following the development of standardized technology, they are now becoming well-established prognostic and predictive markers in patients with breast, colon and prostate cancer. While they represent a real-time noninvasive test, their use in diagnostics has seldom been reported. We report their use to help diagnose an indeterminate pancreatic mass. The use of an automated circulating tumor cell platform as described is likely to have utility as an aid to differential diagnosis, although larger studies will be required to ascertain its positive or negative predictive value. PMID- 20021214 TI - Tumor metabolism of lactate: the influence and therapeutic potential for MCT and CD147 regulation. AB - Tumor metabolism consists of complex interactions between oxygenation states, metabolites, ions, the vascular network and signaling cascades. Accumulation of lactate within tumors has been correlated with poor clinical outcomes. While its production has negative implications, potentially contributing to tumor progression, the implications of the ability of tumors to utilize lactate can offer new therapeutic targets for the future. Monocarboxylate transporters (MCTs) of the SLC16A gene family influence substrate availability, the metabolic path of lactate and pH balance within the tumor. CD147, a chaperone to some MCT subtypes, contributes to tumor progression and metastasis. The implications and consequences of lactate utilization by tumors are currently unknown; therefore future research is needed on the intricacies of tumor metabolism. The possibility of metabolic modification of the tumor microenvironment via regulation or manipulation of MCT1 and CD147 may prove to be promising avenues of therapeutic options. PMID- 20021219 TI - Fate of recombinant adeno-associated viral vector genomes during DNA double strand break-induced gene targeting in human cells. AB - Recombinant vectors based on adeno-associated virus (rAAV) are promising tools to specifically alter complex genomes through homologous recombination (HR)-based gene targeting. In a therapeutic setting, an AAV donor vector will recombine with a mutant target locus in order to correct the mutation directly in the genome. The low frequency of HR in mammalian cells can be significantly improved by insertion of a DNA double-strand break (DSB) into the target locus through expression of a site-specific endonuclease. Here, we have scrutinized the fate of rAAV vector genomes during DSB-induced gene targeting and assessed the targeting frequency and the targeting ratio as a risk-benefit indicator. In various human cell lines carrying a mutated enhanced green fluorescent protein locus with a recognition site for the homing endonuclease I-SceI, rAAV-transduced cells were assayed by flow cytometry and by quantitative allele-specific polymerase chain reaction to assess HR and unspecific integration events. Under optimal conditions gene-targeting frequencies of 65% and targeting ratios of 2:1 were achieved, that is, more gene correction than unspecific integrations. The gene-targeting frequency was highly dependent on rAAV vector design, the cell line, and on the presence of a DSB in the target locus. Although expression of I-SceI led to a significant increase in gene targeting, it did not augment unspecific integration. In conclusion, our results reveal the side effects associated with rAAV-mediated gene targeting, but also its great potential for precise genome engineering in a therapeutic context. PMID- 20021221 TI - Introduction to the Special Issue on Non-oncological Uses of Hyperthermia. PMID- 20021220 TI - Lymphadenoma: case report of a rare salivary gland tumor in childhood. AB - Lymphadenoma of the salivary gland is a rare benign tumor with only 11 reported cases in the English language literature, most of which have occurred in adults. We report a case of a lymphadenoma occurring in the parotid gland of a 15-year old girl. The tumor was composed of variably sized cystic cavities within abundant reactive lymphoid tissue. The cystic spaces were filled with eosinophilic secretions with occasional histiocytes. Many of these features were also apparent on cytologic preparations. The cysts were lined by epithelium lacking atypia and showed luminal and abluminal differentiation both by immunohistochemistry and by electron microscopy. Tumor cells were not cycling as determined by MIB1 immunostaining, and the tumor karyotype was normal. This is only the second case to be reported in the pediatric age group. Ultrastructural features and karyotype analysis are reported for the first time. Although this tumor is rarely encountered by pediatric pathologists, awareness of its existence is important to distinguish it from possible malignant mimics, such as lymphoepithelial carcinoma and metastatic mucoepidermoid carcinoma in a lymph node. PMID- 20021222 TI - Heat shock proteins and immunity: application of hyperthermia for immunomodulation. AB - Heat shock proteins (HSPs) play an important role as 'endogenous danger signals' in the immune surveillance system. Extracellular HSPs released from damaged cells can stimulate professional antigen-presenting cells, followed by cytokine release and expression of cell surface molecules. In addition to such activity stimulating innate immunity, extracellular HSPs can promote the cross presentation of HSP-bound peptide antigens to MHC class I molecules in dendritic cells, leading to efficient induction of antigen-specific cytotoxic T lymphocytes. The roles of HSPs stimulating both innate immunity and adaptive immunity can explain at least in part the molecular mechanism by which thermal stress bolsters the host immune system. In the present review, we present novel aspects of the roles of HSPs in immunity and discuss the therapeutic application of hyperthermia for immunomodulation. PMID- 20021223 TI - Heat shock protein 72 protects insulin-secreting beta cells from lipopolysaccharide-induced endoplasmic reticulum stress. AB - PURPOSE: Hyperthermia-induced activation of stress response proteins allows cells to withstand metabolic insults. In this study we set out to determine whether insulin secretion by pancreatic beta cells was affected by the acute inflammatory response, systemic inflammation-induced hyperglycaemia, and whole-body hyperthermia. Given that systemic-inflammation induces ER stress, we further examined whether hyperthermia can attenuate the extent of LPS-induced ER stress. MATERIALS AND METHODS: Rats were randomised and divided into three treatment groups. Control rats received a 0.9% NaCl solution. Rats in the lipopolysaccharide (LPS) group received 7.5 mg of LPS/kg. Rats in the whole-body hyperthermia (WBH) + LPS group were exposed to 42 degrees C for 15 min, followed by injection with 7.5 mg of LPS/kg after 48 h. Glucose-potentiated insulin release and extent of ER stress were measured in beta cells. RESULTS: LPS inhibited glucose-induced insulin release from islet cells and induced the expression of Bip/GRP78, XBP-1, and CHOP transcripts. The inhibition of glucose induced insulin release and induction of ER stress proteins by LPS was attenuated by WBH. CONCLUSIONS: Our findings suggest that LPS-induced systemic inflammation decreased insulin release due to the effects of ER stress proteins on insulin secretion. Furthermore, the induction of ER stress proteins was prevented by pretreating rats with WBH. This may suggest that inhibiting the induction of ER stress proteins through WBH can restore insulin release in various disease states. PMID- 20021224 TI - Expression and function of stress (heat shock) proteins in gastrointestinal tract. AB - This review will focus on gastrointestinal mucosal protection against cytotoxic agents and cellular stress mainly from the viewpoint of expression and function of heat shock proteins, in their role of 'molecular chaperones', as internal cytoprotectants. Also, recently identified target molecules of heat shock protein in damaged gastric mucosal cells are introduced. Elucidation of such stress responses and repairing process of damaged protein by heat shock proteins in the gastrointestinal mucosa may provide a better understanding for the mechanisms of cytoprotection and cellular repair. In addition, these findings in post-genomic level may provide new strategies for the therapy of gastrointestinal disorders. PMID- 20021225 TI - Heat shock proteins in neurodegenerative diseases: pathogenic roles and therapeutic implications. AB - Neurodegenerative diseases including amyotrophic lateral sclerosis, Parkinson's disease, Alzheimer's disease, and polyglutamine (polyQ) diseases are thought to be caused by protein misfolding. Heat shock proteins (HSPs), which function mainly as molecular chaperones, play an important role in the folding and quality control of proteins. The histopathological hallmark of neurodegenerative diseases is accumulation and/or inclusions of the disease-causing proteins in residual neurons in targeted regions of the nervous system. The inclusions combine with many components of molecular chaperone pathways and ubiquitin-proteasome, raising the possibility that misfolding and altered degradation of mutant proteins may be involved in the pathogenesis of neurodegenerative diseases. Overexpression of HSPs has been reported to reduce the number and size of inclusions and accumulation of disease-causing proteins, and ameliorate the phenotypes in neuronal cell and mouse models. Hsp90 inhibitors also exert therapeutic effects through selective proteasome degradation of its client proteins. Elucidation of its pathophysiology using animal models has led to the development of disease modifying drugs, i.e., Hsp90 inhibitor and HSP inducer, which inhibit the pathogenic process of neuronal degeneration. These findings may provide the basis for development of an HSP-related therapy for neurodegenerative diseases. PMID- 20021226 TI - A novel combination of mild electrical stimulation and hyperthermia: general concepts and applications. AB - This review discusses the basic concepts, effects and applications of hyperthermia and mild electrical stimulation (MES) using low-intensity direct current. It also proposes a novel combinatorial use of MES and hyperthermia, and briefly outlines the rationale and the effects of MES and hyperthermia combination treatment on certain diseases (diabetes, hepatic ischaemia/reperfusion injury and gastric ulcer). The integrated modalities of MES and hyperthermia might find therapeutic applications to stress-induced diseases and intractable diseases of dysregulated signalling pathways. PMID- 20021227 TI - Protective role of HSF1 and HSP70 against gastrointestinal diseases. AB - PURPOSE: It is well known that heat shock proteins (HSPs) are induced by various stressors in order to confer protection against such stressors. Since stressor induced tissue damage is involved in various diseases, especially gastrointestinal diseases, it has been thought that HSP-inducers are therapeutically beneficial for these diseases. Indirect lines of evidence suggest that HSPs provide a major protective mechanism against irritant-induced gastric lesions. However, no direct evidence exists to support this notion. On the other hand, inflammatory bowel disease (IBD) involves infiltration of leukocytes into intestinal tissue, resulting in intestinal damage. Pro-inflammatory cytokines and cell adhesion molecules (CAMs) play important roles in this infiltration of leukocytes. The roles of heat shock factor 1 (HSF1, a transcription factor for hsp genes) and heat shock proteins (HSPs) in development of IBD are unclear. In this paper, we reviewed our recent work on the role of HSPs in pathogenesis of gastric lesions and IBD by use of HSF1-null mice and transgenic mice expressing Hsp70. CONCLUSION: This study provides the first genetic evidence that HSF1 and Hsp70 play a role in protecting against both irritant-induced gastric lesions and IBD-related colitis. The aggravation of irritant-induced gastric lesions in HSF1 null mice is due to their inability to up-regulate Hsp70, leading to apoptosis. On the other hand, this protective role of Hsp70 against colitis seems to involve various mechanisms, such as suppression of expression of pro-inflammatory cytokines and CAMs, and cell death. PMID- 20021229 TI - Liquid crystalline Pluronic 105 pharmacogels as drug delivery systems: preparation, characterization, and in vitro transdermal release. AB - In this study, we report the results of our investigations on the percutaneous permeation profiles of Diclofenac sodium, Paracetamol, Propanolol hydrochloride, and alpha-Tocopherol from the different lyotropic liquid crystalline phases obtained by Pluronic P105/water mixtures, in order to understand if the particular assembly shown in the formulations could influence the delivery across the skin. Recent studies have focused on the Pluronic liquid crystalline phases to evaluate the potential use of these phases in drug delivery, but no comparative investigation has been yet performed on the drug permeation from the different liquid crystalline phases obtained by the same Pluronic surfactant. The cubic, hexagonal, and lamellar mesophases (loading the above-mentioned drug) were characterized by Deuterium Nuclear Magnetic Resonance spectroscopy and Polarized Optical Microscopy observations. Results revealed that the liquid crystalline gel microscopic structure obtained in the different formulations drastically affects the drug percutaneous availability. As a consequence these systems could be proposed as novel transdermal drug delivery systems. PMID- 20021230 TI - Gastroretentive particles formulated with thiomers: development and in vitro evaluation. AB - The objective of this study is to develop and evaluate gastroretentive particulate delivery systems using Riboflavin-5'-monophosphate sodium salt dihydrate (RF5'PNa) as model drug. Poly(acrylic acid)-cysteine and chitosan-4 thiobuthylamidine were evaluated and compared as anionic and cationic polymers for gastroretentive particles. Permeation studies were performed with freshly excised stomach mucosa from rats. Polymers and combination with glutathione were evaluated for permeation enhancing properties. Furthermore, particles were prepared by air jet milling and characterized. Permeation studies showed that the apparent permeability coefficients for RF5'PNa with thiomers and glutathione are 1.511-fold and 2.354-fold higher than control, respectively. It can be seen from the results glutathione in combination with thiomers has a significant influence for increasing permeation. Poly(acrylic acid)-cysteine and chitosan-4 thiobuthylamidine particles demonstrated a mean diameter of 336.5 +/- 16.5 and 396.3 +/- 17.0 nm and zeta potential of -19.98 +/- 1.015 and 27.15 +/- 0.500 mV, respectively. The drug loading of Poly(acrylic acid) particles was significantly higher than chitosan particles. The release rate of RF5'PNa from the thiolated particles was slower compared with unmodified particles. Moreover, thiolated particles showed higher mucoadhesive properties compared to unmodified particles. Overall, thiolated particles of both anionic and cationic polymers had improved mucoadhesive and controlled release properties. Therefore, they could be promising for gastroretentive delivery systems. PMID- 20021231 TI - Different serum enzyme levels are required to rescue the various systemic features of the mucopolysaccharidoses. AB - Mucopolysaccharidoses (MPSs) are lysosomal storage disorders characterized by progressive accumulation of glycosaminoglycans (GAGs) in various tissues. Enzyme replacement therapy (ERT) for several MPSs is available to date. However, the efficacy of ERT is limited, in particular in compartments such as bone, cartilage, the brain, and the eyes. We selected a rodent model of an MPS, with no central nervous system storage, to study the impact, on systemic features of the disease, of various stable levels of exogenous enzymes produced by adeno associated viral vector (AAV)-mediated liver gene transfer. Low levels (6% of normal) of circulating enzyme were enough to reduce storage and inflammation in the visceral organs and to ameliorate skull abnormalities; intermediate levels (11% of normal) were required to reduce urinary GAG excretion; and high levels (>or=50% of normal) rescued abnormalities of the long bones and motor activity. These data will be instrumental to design appropriate clinical protocols based on either enzyme or gene replacement therapy for MPS and to predict their impact on the pathological features of MPS. PMID- 20021233 TI - Laparoscopic versus robotic pyeloplasty: man versus machine. AB - With all its different presentations and etiologies, ureteropelvic junction obstruction has been a topic for much research and debate. For several decades, the 'gold standard' of treatment was unequivocally an Anderson-Hynes dismembered pyeloplasty. Various surgical modifications and minimally invasive alternatives have been studied. It was not until the last two decades that laparoscopic and robotic approaches have threatened to supplant the classic open approach as the preferred surgical treatment option. While the debate between the laparoscopic and robotic approaches has been a heavily contested one, it has rarely been founded on prospective, well-matched evidence. We review the existing literature and present our perspective on the clinical, academic and economic aspects of this contest between man and machine. PMID- 20021232 TI - Changes in adeno-associated virus-mediated gene delivery in retinal degeneration. AB - Gene therapies for retinal degeneration have relied on subretinal delivery of viral vectors carrying therapeutic DNA. The subretinal injection is clearly not ideal as it limits the viral transduction profile to a focal region at the injection site and negatively affects the neural retina by detaching it from the supportive retinal pigment epithelium (RPE). We assessed changes in adeno associated virus (AAV) dispersion and transduction in the degenerating rat retina after intravitreal delivery. We observed a significant increase in AAV-mediated gene transfer in the diseased compared with normal retina, the extent of which depends on the AAV serotype injected. We also identified key structural changes that correspond to increased viral infectivity. Particle diffusion and transgene accumulation in normal and diseased retina were monitored via fluorescent labeling of viral capsids and quantitative PCR. Viral particles were observed to accumulate at the vitreoretinal junction in normal retina, whereas particles spread into the outer retina and RPE in degenerated tissue. Immunohistochemistry illustrates remarkable changes in the architecture of the inner limiting membrane, which are likely to underlie the increased viral transduction in diseased retina. These data highlight the importance of characterizing gene delivery vectors in diseased tissue as structural and biochemical changes can alter viral vector transduction patterns. Furthermore, these results indicate that gene delivery to the outer nuclear layer may be achieved by noninvasive intravitreal AAV administration in the diseased state. PMID- 20021234 TI - Implantable cardioverter-defibrillator therapy in congenital heart disease. PMID- 20021236 TI - TissueMend. PMID- 20021237 TI - Permanent or temporary IVC filtration with a novel double-ring anchoring technology optional nitinol filter. AB - Permanent or temporary implantation of inferior vena cava filters for protection against pulmonary embolism is well established. There have been numerous devices developed for this purpose, each of which has proprietary design considerations that affect performance and potentially impose limitations with regard to positioning, efficacy and risk profile. This article describes a recently developed, unique inferior vena cava filter design that employs a separated filtration component and a novel double-ring anchoring system that allows intraprocedural capture and repositioning for optimized placement. In addition, early experience suggests easy removal when desired, a high rate of filtration success and excellent caval patency. PMID- 20021238 TI - Endonasal phototherapy with Rhinolight for the treatment of allergic rhinitis. AB - Allergic rhinitis, although not life threatening, significantly affects the quality of the patient's daily life. The three major steps in the treatment of the condition are avoidance of allergens, treatment of symptoms (in particular, antihistaminics and topical nasal corticosteroids) and specific immunotherapy. Avoidance of the allergen is usually not possible and symptom relief is often limited, despite the availability of a number of pharmacological options. Specific immunotherapy demands a high level of cooperation on the part of the patient for at least 3 years. Endonasal phototherapy with the Rhinolight device (Rhinolight Ltd, Szeged, Hungary) for the treatment of immunoglobulin E-mediated allergic rhinitis is a new option that utilizes the immunosuppressive effects of UV radiation. The method directs a combination of UV-B (5%), UV-A (25%) and visible light (70%) into the nasal cavity, and its effectiveness has been demonstrated in one double-blind, placebo-controlled study. The results of additional studies have been presented at various medical conferences and in abstracts. Reports in the literature confirm that phototherapy is a well established and successful treatment of atopic dermatitis and other skin diseases. PMID- 20021239 TI - Development of bioactive conducting polymers for neural interfaces. AB - Bioelectrodes for neural recording and neurostimulation are an integral component of a number of neuroprosthetic devices, including the commercially available cochlear implant, and developmental devices, such as the bionic eye and brain machine interfaces. Current electrode designs limit the application of such devices owing to suboptimal material properties that lead to minimal interaction with the target neural tissue and the formation of fibrotic capsules. In designing an ideal bioelectrode, a number of design criteria must be considered with respect to physical, mechanical, electrical and biological properties. Conducting polymers have the potential to address the synergistic interaction of these properties and show promise as superior coatings for next-generation electrodes in implant devices. PMID- 20021240 TI - Scientific evidence for the use of modern patellofemoral arthroplasty. AB - Patellofemoral arthroplasty has been utilized as a treatment for isolated patellofemoral arthritis for more than 30 years. However, the use of this procedure remains controversial, as many surgeons prefer to use total knee arthroplasty, even for isolated patellofemoral arthritis. While historically, the results with this procedure have been inconsistent, recent developments in prosthesis design and surgical indications have improved the outcomes of patellofemoral arthroplasty. Potential advantages of patellofemoral arthroplasty include a less invasive approach, less bone resection, less tissue destruction, shorter operative time, less blood loss, shorter rehabilitation, and more normal knee kinematics. However, proper indications and surgical technique are crucial in order to obtain optimal results. Some future modifications have the potential to further improve the outcomes of the procedure, although additional investigations are needed to further explore some of these aspects. This report will describe current knowledge regarding the indications and contraindications for patellofemoral arthroplasty, present the results and complications of this procedure, discuss alternative treatments for patellofemoral disease, and explore future directions for arthroplasty of the patellofemoral compartment. PMID- 20021241 TI - Why do some promising brain-stimulation devices fail the next steps of clinical development? AB - Interest in techniques of noninvasive brain stimulation (NIBS) has been growing exponentially in the last decade. Recent studies have shown that some of these techniques induce significant neurophysiological and clinical effects. Although recent results are promising, there are several techniques that have been abandoned despite positive initial results. In this study, we performed a systematic review to identify NIBS methods with promising preliminary clinical results that were not fully developed and adopted into clinical practice, and discuss its clinical, research and device characteristics. We identified five devices (transmeatal cochlear laser stimulation, transcranial micropolarization, transcranial electrostimulation, cranial electric stimulation and stimulation with weak electromagnetic fields) and compared them with two established NIBS devices (transcranial magnetic stimulation and transcranial direct current stimulation) and with well-known drugs used in neuropsychiatry (pramipexole and escitalopram) in order to understand the reasons why they failed to reach clinical practice and further steps of research development. Finally, we also discuss novel NIBS devices that have recently showed promising results: brain ultrasound and transcranial high-frequency random noise stimulation. Our results show that some of the reasons for the failure of NIBS devices with promising clinical findings are the difficulty to disseminate results, lack of controlled studies, duration of research development, mixed results and lack of standardization. PMID- 20021242 TI - Imaging as a surveillance tool in rectal cancer. AB - Despite advances in diagnosis and treatment, half of patients with treated rectal cancer will die owing to recurrent disease. There is no evidence of benefit on survival from an intensive surveillance program, even if presymptomatic recurrent disease is detected. The aim of this article is to review the results described for the different imaging techniques in diagnosing rectal cancer recurrence in different sites and to discuss their relative clinical impact. The sensitivity of imaging techniques is related to the performance of the machines and the site being examined. Computed tomography is the most used technique owing to its availability, speed, panoramic images and ease of use, while MRI of the pelvis and the liver produces the highest resolution, sensitivity and specificity in these anatomical areas. Owing to its high cost, [(18)F] fluorodeoxyglucose-PET should be used as a third-level examination, a 'problem-solving' method when the site of recurrence is unknown or to rule out other possible sites of recurrence before a second surgery, and, finally, because it offers the possibility to investigate the whole body. The follow-up must be designed for individual patients, taking into account a number of factors. In the near future, whole-body imaging, probably by MRI, that is free from radiation will become the method of choice for screening for recurrent disease. PMID- 20021243 TI - Computer-aided manufacturing technologies for guided implant placement. AB - Implant treatment increasingly focuses on the reduction of treatment time and postoperative impairment. The improvement of 3D dental diagnosis by ConeBeam computed tomography allows detailed preparation for the surgical placement of dental implants under prosthetic considerations. While the first generation of implant planning software used high-contrast multislice computed tomography, software that has been specifically designed for ConeBeam computed tomography is now available. Implant placement can be performed using surgical guides or under the control of optical tracking systems. Surgical guides are more commonly used in private office owing to their availability. The accuracy for both techniques is clinically acceptable for achieving implant placement in critical anatomical indications. When using prefabricated superstructures and in flapless surgery, special abutments or an adjusted workflow are still necessary to compensate misfits of between 150 and 600 microm. The proposition to ensure proper implant placement by dentists with limited surgical experience through the use of surgical guides is unlikely to be successful, because there is also a specific learning curve for guided implant placement. Current and future development will continue to decrease the classical laboratory-technician work and will integrate the fabrication of superstructures with virtual treatment planning from the start. PMID- 20021244 TI - Surfaces to control tissue adhesion for osteosynthesis with metal implants: in vitro and in vivo studies to bring solutions to the patient. AB - For internal fracture-fixation, metal currently remains the material of choice, since it provides strength for bone fragment support, good ductility for presurgical contouring and has been shown extensively to be biopassive. For decades, the application of metal internal fixators has proven undoubtedly successful and is deemed by many as the greatest advance in orthopedic medicine to date. However, based on this unrivalled success, newer and more challenging applications for metal internal fixators have emerged. For instance, given the large increase in the occurrence of these procedures in children and the different mechanical and biological requirements based on anatomical site of implantation, the functional requirements of metal implants have become increasingly more demanding. Given this changing demand for metal internal fixators, a more pragmatic application approach is necessary. Therefore, current metal internal fixator-related orthopedic research is based on defining specific cell and tissue responses to materials both in vitro and in vivo, as well as methods to empirically facilitate implantation site-specific tissue responses. This review discusses current knowledge from both the author's as well as others' laboratories pertaining to cell- and tissue-specific responses to metal internal fixation materials, with specific emphasis on a surface microtopographical approach to alleviating removal-related morbidity. The review also describes the 'effective roughness spectrum' hypothesis for control of cell surface integration. PMID- 20021245 TI - Recent advances in dermal wound healing: biomedical device approaches. AB - Successful repair of wounds and tissues remains a major healthcare and biomedical challenge in the 21st Century. In particular, chronic wounds often lead to loss of functional ability, increased pain and decreased quality of life, and can be a burden on carers and health-system resources. Advanced healing therapies employing biological dressings, skin substitutes, growth factor-based therapies and synthetic acellular matrices, all of which aim to correct irregular and dysfunctional cellular pathways present in chronic wounds, are becoming more popular. This review focuses on recent advances in biologically inspired devices for wound healing and includes a commentary on the challenges facing the regulatory governance of such products. PMID- 20021247 TI - Re-emergence of the progenitors of a multidrug-resistant outbreak strain of Mycobacterium tuberculosis among the post-outbreak case patients. AB - BACKGROUND. The progenitors of multidrug-resistant tuberculosis (MDR-TB) outbreak strains might evolve into new outbreak strains. We hypothesized that these strains could re-emerge among post-outbreak patients with TB and must thus be tracked. METHODS. To identify the progenitors of the outbreak strain, we first determined the precise IS6110 genomic insertion locations of a Haarlem3 strain that caused a severe MDR-TB outbreak in Tunisia. Next, we searched by polymerase chain reaction for these outbreak-specific IS6110 transposition sites in all the Haarlem3 post-outbreak isolates recovered in the epidemic region. RESULTS. By analyzing the distribution of the outbreak-specific IS6110 transposition sites, we were able to trap, among isolates recovered from post-outbreak new patients, drug-susceptible and drug-resistant isolates that are likely to represent the very close progenitors of the outbreak strain. Mycobacterial interspersed repetitive units-variable number of tandem repeats typing and sequential accumulation of rifampicin resistance-associated rpoB mutations further confirmed the identity of the outbreak's progenitor strains. CONCLUSIONS. The data of the current study show that the progenitors of an MDR-TB outbreak strain could re emerge among post-outbreak TB cases. We provide an IS6110 insertion site-based approach to better trace back the events preceding the emergence of MDR-TB outbreaks, irrespective of the availability of a pre-outbreak strain collection. PMID- 20021248 TI - Nasal-associated lymphoid tissue is not an absolute requirement for the induction of rat tear IgA antibody responses. AB - PURPOSE/AIM OF STUDY: The purpose of this work was to determine whether rat nasal associated lymphoid tissue is required for the induction of tear IgA responses. MATERIALS AND METHODS: Particulate antigen in the form of DNP-BSA encapsulated in cationic microparticles was applied topically to the eyes (ocular topically) of rats that had the nasolacrimal ducts temporarily plugged with chromic gut suture material. Eye washes and serum were monitored for development of antigen specific IgA and IgG, respectively. To track the particulate uptake, fluorescent latex beads were applied topically to the eyes of plugged and unplugged animals. The nasal-associated lymphoid tissue and the draining lymph nodes were then examined for the presence of the fluorescent beads. RESULTS: It was found that the chromic gut suture was effective in blocking the passage of antigen into the nasopharyngeal cavity for at least 24 hr. Tear antigen-specific IgA levels found in the eyes of plugged animals were not significantly lower from those of unplugged animals. Serum IgG antibody levels were also similar between the two groups. In animals with plugged nasolacrimal ducts, fluorescent beads were found predominately in the superficial cervical lymph nodes, which have been shown to drain the surface of the eye. CONCLUSIONS: These results indicate that particulate antigen can be taken up by the conjunctiva and transported to the draining lymph nodes, showing that antigen does not need to access nasal associated lymphoid tissue to induce tear IgA antibody responses. PMID- 20021249 TI - Reverse geometry rigid gas permeable contact lens wear reduces high-order aberrations and the associated symptoms in post-LASIK patients. AB - PURPOSE: This study evaluated the efficacy of custom-made reverse geometry rigid gas permeable (RGP) contact lenses in the control of symptoms related to increased wavefront aberrations after LASIK correction of myopia. MATERIALS AND METHODS: Twenty-eight myopes treated with LASIK were assessed for post-LASIK symptoms (declined night vision, glare, halos and/or monocular diplopia) and corneal topography. A set of RGP contact lenses was selected as trial lenses based on the matching between the lens and the patient's corneal topography. If the fluorescein pattern between the trial lens and the corneal surface showed a central alignment, a mid-peripheral bearing and peripheral clearance and if the patient felt comfortable with the lens wear, a duplicated lens was ordered for the patient. Otherwise, a lens satisfied with corneal topography of the patient and clinical judgment was used. Visual acuity (VA) and wavefront aberrations were measured after the lens wear. RESULTS: The custom-made lenses well matched the host cornea with no dislodgment in all subjects. The post-LASIK eyes showed an increase of 1 Snellen line in VA (from 0.90 +/- 0.33 to 1.11 +/- 0.24) and a significant improvement in all symptoms after the lens wear. The total higher order aberration is significantly reduced after the lens wear with at least 70% reduction in either spherical aberration, coma, third or fourth aberration but only 33% reduction in the fifth aberration. CONCLUSIONS: Reverse geometry RGP lenses with the design based on individual topographic data can improve visual performance of post-LASIK eyes by reducing higher order aberrations. PMID- 20021250 TI - Efficacy of topical bevacizumab for inhibiting growth of impending recurrent pterygium. AB - PURPOSE: To evaluate clinical outcome(s) and complication(s) of topical bevacizumab (avastin) in patients with impending recurrent pterygium. PATIENTS AND METHODS: This prospective clinical trial included 54 consecutive patients (54 eyes), who underwent pterygium surgery with bare sclera and mitomycin, and who were diagnosed with impending recurrent pterygium. Of the 54 patients, 26 received eye drops containing bevacizumab (5 mg/ml) twice a day and betamethasone four times daily for 1 week. In the other 28 patients, betamethasone alone was administered four times daily for 1 week. Follow-up times were 3-6 mos. RESULTS: All patients in both groups failed, i.e., fibrovascular tissue (pterygia) eventually extended onto the cornea. However, in patients receiving topical bevacizumab, the mean progression of fibrovascular tissue extension during the first week, first month, and third month of follow-up was significantly less than that in the control group (p < 0.01). The mean duration for invasion of cornea in study group patients was significantly longer than that for control group patients (p < 0.01). CONCLUSION: Short-term use of topical bevacizumab seems to be a safe and effective means for delaying recurrence of impending recurrent pterygia. PMID- 20021251 TI - Lens cadmium, lead, and serum vitamins C, E, and beta carotene in cataractous smoking patients. AB - PURPOSE: To investigate the association between smoking and subcapsular cataract and blood levels of cadmium and lead, and serum levels of vitamins C, E, and beta carotene in the middle-age male population. SUBJECTS AND METHODS: The present study comprised 60 cataractous male patients aged 44-55 years who attended Mansoura University Ophthalmic Center. They were divided into two groups: the first group was comprised of 15 patients who had never smoked before with minimal exposure to cigarette smoking (control group), while the second group was comprised of 45 age and body mass index-matched smokers. Blood cadmium and lead concentrations were measured and their levels in the cataractous lenses were also estimated. The levels of some antioxidants, such as serum vitamins C, E, and beta carotene, were determined. RESULTS: Cadmium had a statistically significant higher concentration in blood and lenses of smokers compared with that of non smokers (p < 0.0001). There was a significantly high accumulation of cadmium in the lenses of cataractous smokers reaching about a four-fold increase in heavy smokers (15.4 +/- 0.4 micromol/g) and nearly a three-fold increase in light smokers (10.1 +/- 0.4 micromol/g) when compared to that of nonsmokers (3.7 +/- 0.9 micromol/g). Regarding vitamins E, C, and beta carotene, a highly significant reduction was observed in smokers when compared with nonsmokers. There was a highly positive correlation between blood cadmium concentrations and lens cadmium levels, and blood lead concentrations and lens lead levels in heavy smokers. CONCLUSION: Cadmium present in high concentrations among smokers was associated with the presence of cataracts, through induction of oxidative stress as evidenced by reduction of levels of some antioxidant studied in this work, such as vitamins E, C, and beta carotene. Thus, education of health care professionals and the public about the cataractogenic effect of cadmium is mandatory. Intake of various antioxidants may be helpful in reducing the risk of cataract formation. PMID- 20021252 TI - Variable clinical spectrum of the myocilin Gln368X mutation in a Dutch family with primary open angle glaucoma. AB - PURPOSE: To describe the clinical phenotype in a family with primary open angle glaucoma harboring a p.Gln368X mutation in MYOC. MATERIALS AND METHODS: We identified a proband with primary open angle glaucoma and the p.Gln368X MYOC mutation. She and her six siblings were examined clinically, including Heidelberg Retina Tomography II, and venous blood samples were screened for other variants in MYOC, WDR36, OPTN, and CYP1B1. RESULTS: Four individuals showed the p.Gln368X MYOC mutation, no other genetic variations were assessed. Two of these four siblings had glaucomatous optic disc changes with corresponding visual field losses and abnormal Heidelberg Retina Tomography results by the Moorfields regression analysis, one had abnormal results by the Moorfields regression analysis but no visual field loss, and one showed no glaucomatous signs or symptoms at all. These findings did not correlate with the age of the affected individuals. CONCLUSION: In the primary open angle glaucoma family described here, we documented a wide range in clinical symptoms, demonstrating a highly variable penetrance of the MYOC p.Gln368X mutation. PMID- 20021253 TI - TGF-beta2 in human retinal pigment epithelial cells: expression and secretion regulated by cholinergic signals in vitro. AB - PURPOSE: To investigate the (i) effect(s) of cholinergics on the expression and secretion of transforming growth factor (TGF)-beta(2) in human retinal pigment epithelium (RPE) and (ii) mechanism of action of atropine in the treatment of myopia. MATERIALS AND METHODS: The RPE cell line, D407, was (i) treated with carbachol (10 microM), (ii) treated with atropine (10 nM-100 microM), or (iii) pre-treated with atropine (10 nM-100 microM) and then exposed to carbachol (10 microM). A no-treatment group served as control. Expression of TGF-beta(2), after stimulation at different time points (2, 4, 8, 16, 24, and 48 hr), was measured by RT-PCR and Western blot analysis. Secretion of TGF-beta(2) was determined by ELISA. RESULTS: Carbachol induced a time-dependent increase in the levels of TGF beta(2) mRNA and protein in the cytoplasm (p < 0.001). ELISA assays showed a time dependent increase in levels of TGF-beta(2) protein in the supernatant with carbachol treatment (p < 0.001). There was no change of TGF-beta(2) in the cytoplasm or supernatant with atropine alone (p > 0.05). The increased expression and secretion of TGF-beta(2) caused by carbachol were suppressed by atropine (in the range of 10 nM-100 microM) when compared to treatment with carbachol alone (p < 0.001). The stimulating effect of 10 microM carbachol was inhibited completely by 100 microM atropine. CONCLUSIONS: In RPE cells, atropine inhibits the expression and secretion of TGF-beta(2) by blocking the muscarinic acetylcholine receptor (mAChR), which may control the development of myopia. PMID- 20021254 TI - Inhibitory effects of arresten on bFGF-induced proliferation, migration, and matrix metalloproteinase-2 activation in mouse retinal endothelial cells. AB - PURPOSE: The potential role of arresten (alpha1(IV)NC1) as an endogenous angiogenesis inhibitor in the prevention of bFGF mediated retinal angiogenesis and regulation of matrix metalloproteinase-2 activation has not been explored. METHODS: Mouse retinal endothelial cells (MREC) were cultured on type IV collagen and treated with basic fibroblast growth factor (bFGF) alone or in the presence of arresten at concentrations ranging from 1 to 10 microg/ml. The proliferation of MRECs were evaluated using 3(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) colorimetric assay, and bFGF stimulated endothelial cell migration was assessed using Boyden chamber. Expression of matrix metalloproteinase-2 (MMP-2) was assessed by reverse transcription polymerase chain reaction (RT-PCR) analysis using RNA isolated from MRECs. Secretion and activation of MMP-2 in arresten-treated conditioned MREC growth medium was determined by gelatin zymography and Western blotting. RESULTS: Different doses of bFGF induced MREC proliferation was significantly inhibited upon arresten treatment (P < 0.005). The bFGF-induced migration was significantly inhibited by arresten at 1 and 10 microg/ml concentrations (P < 0.01). The bFGF stimulated expression of MMP-2 mRNA and secretion of MMP-2 in MREC was not affected and interestingly activation of MMP-2 was suppressed by arresten in a dose and time dependent manner. CONCLUSIONS: Inhibitory effects of arresten on proliferation, migration and MMP-2 activation but not on expression and secretion of MMP-2 in MREC; this early work with arresten supports potential therapeutic action in retinal neovascularization dependent disorders. PMID- 20021255 TI - Rabbit retinal neovascularization induced by latex angiogenic-derived fraction: an experimental model. AB - PURPOSE: To create a retinal neovascularization experimental model using intravitreal injection of microspheres loaded with latex-derived angiogenic fraction. METHODS: Thirty-two albino New Zealand rabbits, divided in 4 groups of 8 animals, were enrolled in this study. Rabbits in groups I, II, and III received one intravitreal injection of PLGA (L-lactide-co-glycolide) microspheres with 10, 30, and 50 microg of latex-derived angiogenic fraction into their right eyes, respectively, and group IV received 0.1 ml of microspheres without the angiogenic fraction. Weekly follow-up with ophthalmoscopy and fluorescein angiography was performed; the rabbits were sacrificed in the 4th week and their eyes processed for light microscopy. RESULTS: All eyes from group I demonstrated increased retinal vascular tortuosity, observed from 14 days after injection and maintained for 28 days, otherwise without new vessels detection. All group II eyes showed vascular changes similar to group I. Fifty percent of the eyes from group II rabbits developed retinal neovascularization 21 days after injection. All eyes from group III demonstrated significant vascular tortuosity and retinal new vessels 2 weeks after injection, progressing to fibrovascular proliferation and tractional retinal detachment. No vascular changes or retinal new vessels were observed in group IV eyes. Light microscopy confirmed the existence of new vessels previously seen on fluorescein angiography, in retinal sections adjacent to the optic disc, not observed in sections at the same area in the control group. CONCLUSION: Thirty- and 50-microg microspheres containing latex-derived angiogenic fraction injected into the vitreous cavity induced retinal neovascularization in rabbits. PMID- 20021256 TI - Test-retest reliability and inter-ocular symmetry of multi-focal electroretinography in Stargardt disease. AB - PURPOSE: To evaluate test-retest reliability and inter-ocular symmetry of multi focal electroretinography recordings in Stargardt disease and in patients without retinal pathology. METHODS: Multi-focal electroretinography were recorded with continuous fundus monitoring using the VERIS multifocal recording system using an array of 103 hexagons spanning 50 degrees of central retina. Recordings were made sequentially from one eye (test-retest) followed by recordings from the fellow eye. A departure score, expressed as a percentage difference from the first recording (or from the fellow eye), was calculated for each comparison and parameter. RESULTS: Multi-focal electroretinography response topographies were similar within and between eyes for a particular individual and parameter. However, local response variability within and between eyes was significantly higher in Stargardt disease than in patients without retinal pathology for both amplitude and timing parameters. Amplitude variability in Stargardt disease decreased with more peripheral targets and with larger stimuli but never to the levels achieved with normal observers. CONCLUSIONS: Both test-retest reliability and inter-ocular symmetry of multi-focal electroretinography responses were significantly lower in Stargardt disease compared to normal eyes, a finding attributable primarily to unsteady fixation. The greater variability between eyes in Stargardt disease may also reflect pathological differences in the topography of retinal function. This inherent variability must be taken into consideration if the multi-focal electroretinography is to be sufficiently sensitive to reliably detect small differences in retinal function. PMID- 20021257 TI - Mutations in RPGR and RP2 of Chinese patients with X-linked retinitis pigmentosa. AB - PURPOSE: To identify mutations in the RPGR and RP2 genes from Chinese families with X-linked retinitis pigmentosa (XLRP). MATERIALS AND METHODS: DNA fragments encompassing coding exons and adjacent intronic regions of RPGR and RP2-were analyzed by cycle sequencing. RESULTS: Three mutations (ORF15 + 483_484delGA, ORF15 + 652_653delAG, and ORF15 + 650_653delAGAG) in RPGR were identified in four families with XLRP, while two mutations (c.353G>A and c.103_1053del) in RP2 were detected in two families with retinitis pigmentosa (RP) and high myopia. CONCLUSIONS: Our results expand the frequency and spectrum of mutations at RPGR and RP2 as well as their associated clinical phenotypes in Chinese patients. PMID- 20021259 TI - Health outcomes and costs of community mitigation strategies for an influenza pandemic in the United States. AB - BACKGROUND: The optimal community-level approach to control pandemic influenza is unknown. METHODS: We estimated the health outcomes and costs of combinations of 4 social distancing strategies and 2 antiviral medication strategies to mitigate an influenza pandemic for a demographically typical US community. We used a social network, agent-based model to estimate strategy effectiveness and an economic model to estimate health resource use and costs. We used data from the literature to estimate clinical outcomes and health care utilization. RESULTS: At 1% influenza mortality, moderate infectivity (R(o) of 2.1 or greater), and 60% population compliance, the preferred strategy is adult and child social distancing, school closure, and antiviral treatment and prophylaxis. This strategy reduces the prevalence of cases in the population from 35% to 10%, averts 2480 cases per 10,000 population, costs $2700 per case averted, and costs $31,300 per quality-adjusted life-year gained, compared with the same strategy without school closure. The addition of school closure to adult and child social distancing and antiviral treatment and prophylaxis, if available, is not cost effective for viral strains with low infectivity (R(o) of 1.6 and below) and low case fatality rates (below 1%). High population compliance lowers costs to society substantially when the pandemic strain is severe (R(o) of 2.1 or greater). CONCLUSIONS: Multilayered mitigation strategies that include adult and child social distancing, use of antivirals, and school closure are cost-effective for a moderate to severe pandemic. Choice of strategy should be driven by the severity of the pandemic, as defined by the case fatality rate and infectivity. PMID- 20021260 TI - The HGF/c-Met axis synergizes with G-CSF in the mobilization of hematopoietic stem/progenitor cells. AB - As granulocyte-colony-stimulating factor (G-CSF)-induced mobilization of hematopoietic stem/progenitor cells (HSPCs) increases human serum levels of hepatocyte growth factor (HGF), our aim was to investigate the role of HGF and its receptor, c-Met, in the mobilization of HSPC. CD34(+) cells and leukocytes were isolated from the bone marrow (BM) of normal donors and the peripheral blood (PB) of patients mobilized with G-CSF and chemotherapy. Plasma HGF levels were evaluated by ELISA and HGF and c-Met expression by RT-PCR, fluorescence-activated cell sorter (FACS) analysis, and confocal microscopy. Because matrix metalloproteinases (MMPs) facilitate migration across extracellular matrix (ECM) and basement membranes, we also examined expression of MMP-9 and membrane type 1 (MT1)-MMP in hematopoietic cells after HGF stimulation. We found that plasma HGF levels in mobilized (m)PB were higher in patients who are good mobilizers and correlated with their white blood cell (WBC) and CD34(+) cell counts. Moreover, HGF and c-Met expression was significantly higher in mPB CD34(+) cells and leukocytes than in their steady-state BM counterpart cells and was up-regulated by G-CSF. Like G-CSF, HGF increased the secretion of MMP-9 and the expression of MT1-MMP in leukocytes, which was abrogated by the c-Met inhibitor K-252a. This inhibitor also significantly reduced the trans-Matrigel migration of mPB CD34(+) cells toward HGF. Our results suggest that G-CSF-mediated HSPC mobilization occurs in part through the HGF/c-Met axis in HSPC and myeloid cells, eliciting increased production of matrix-degrading enzymes and subsequently facilitating egress of HSPC. PMID- 20021264 TI - In anticipation of 2010. PMID- 20021258 TI - Sensitivity of staurosporine-induced differentiated RGC-5 cells to homocysteine. AB - PURPOSE: Homocysteine is implicated in ganglion cell death associated with glaucoma. To understand mechanisms of homocysteine-induced cell death, we analyzed the sensitivity of the RGC-5 cell line, differentiated using staurosporine, to physiologically-relevant levels of the excitotoxic amino acid homocysteine. METHODS: RGC-5 cells were differentiated 24 hr using 316 nM staurosporine and tested for expression of Thy 1.2 via immunodetection, RT-PCR, and immunoblotting. The sensitivity of staurosporine-differentiated RGC-5 cells to physiological levels of homocysteine (50, 100, 250 microM) and to high levels of homocysteine (1 mM), glutamate (1 mM), and oxidative stress (25 microM:10 mU/ml xanthine:xanthine oxidase) was assessed by TUNEL assay and by immunodetection of cleaved caspase-3. The sensitivity of undifferentiated RGC-5 cells to high (1, 5, and 10 mM) homocysteine was also examined. RESULTS: Undifferentiated RGC-5 cells express Thy 1.2 mRNA and protein. Staurosporine differentiated RGC-5 cells extend neurite processes and express Thy 1.2 after 24 hr differentiation; they express NF-L after 1 and 3 days differentiation. Treatment of staurosporine -differentiated RGC-5 cells with 50, 100, or 250 microM homocysteine did not alter neurite processes nor induce cell death (detected by TUNEL and active caspase-3) to a level greater than that observed in the control (non-homocysteine-treated, staurosporine-differentiated) cells. The 1 mM dosage of homocysteine in staurosporine-differentiated RGC-5 cells also did not induce cell death above control levels, although 18 hr treatment of non differentiated RGC-5 cells with 5 mM homocysteine decreased survival by 50%. CONCLUSIONS: RGC-5 cells differentiated for 24 hr with 316 nM staurosporine project robust neurite processes and are positive for ganglion cell markers consistent with a more neuronal phenotype than non-staurosporine-differentiated RGC-5 cells. However, concentrations of homocysteine known to induce ganglion cell death in vivo and in primary ganglion cells are not sufficient to induce death of RGC-5 cells, even when they are differentiated with staurosporine. PMID- 20021265 TI - Two naturally occurring variants of the serotonin receptor gene HTR3C are associated with nausea in pregnancy. AB - OBJECTIVE: To assess the association between pregnancy-associated symptoms and common single nucleotide polymorphisms (SNPs) in genes known to be involved in the pathogenesis of nausea and vomiting. DESIGN: In a standardized, questionnaire based interview, women selected from a control cohort for association studies were asked retrospectively about nausea and vomiting during their first pregnancy. POPULATION: A total of 593 women who had completed at least one pregnancy and for whom germline DNA was available were selected. METHODS: Eight SNPs in the serotonin receptor genes HTR3A, HTR3B, HTR3C, HTR3D, HTR3E, and NK1R (TACR1) were tested using polymerase chain reaction. The occurrence of nausea and vomiting was correlated with the patients' genotyping results and medical history parameters. RESULTS: Both young age at first pregnancy and positive smoking status were significantly associated with vomiting and nausea during pregnancy. After adjustment for these two parameters, the two SNPs rs6806362 (odds ratio (OR) 1.38 per allele; 95% confidence interval (CI) 1.06-1.79; p = 0.017) and rs6807670 (OR 1.37; 95% CI 1.05-1.79 per allele; p = 0.023) were marginally associated with pregnancy-related nausea. None of the other polymorphisms showed any association. CONCLUSION: Polymorphisms in the subunit gene HTR3C of the serotonin receptor may be involved in the pathogenesis of pregnancy-associated nausea. PMID- 20021266 TI - Cervical tissue changes in women with miscarriage: a morphological and biochemical investigation. AB - OBJECTIVE: To evaluate morphological and inflammatory events in the uterine cervix in first trimester miscarriages. DESIGN: Experimental study. SETTING: Sahlgrenska University Hospital, Gothenburg, Sweden. POPULATION: Nulliparous women with first trimester symptomatic (n = 7) or silent (n = 11) miscarriage scheduled for surgical evacuation and nulliparous women scheduled for surgical termination of first trimester pregnancies (control group n = 11). METHODS: Before evacuation, biopsies were obtained from the cervix. The specimens were either fixed in glutaraldehyde for electron microscopy or snap-frozen in liquid nitrogen and stored at -70 degrees C until analyses of interleukin-8 (IL-8) and matrix metalloproteinases (MMPs). MAIN OUTCOME MEASURES: Ultrastructure of cervical tissue, cervical tissue levels of IL-8, immunohistochemistry of MMP-1, MMP-8, MMP-9 and IL-8. RESULTS: The organization of the cervical collagen framework was deranged, the fibroblasts were reactive and the number of activated mast cells appeared to be increased in specimens from women with miscarriage compared with controls. IL-8 was significantly increased in women with miscarriage. Immunohistochemistry of MMP-1 and MMP-8 did not demonstrate any significant difference between the groups. MMP-9 was significantly lower in specimens from women with symptomatic miscarriage compared to women with silent miscarriage and women in the control group. CONCLUSIONS: An inflammatory-like response takes place in the cervix both in women with symptomatic and silent miscarriage. The intensity of the inflammatory response seems to be similar in the two groups. Therefore, inadequate cervical remodeling does not seem to be the reason why some miscarriages remain silent. PMID- 20021267 TI - Radical excision of rectovaginal endometriosis results in high rate of pain relief - results of a long-term follow-up study. AB - OBJECTIVE: To evaluate the long-term results of radical excision for rectovaginal endometriosis (RVE) with special emphasis on current symptoms and risk factors as regards recurrence. METHODS: A total of 116 patients operated upon because of RVE were offered a clinical follow-up evaluation visit; 60 (52%) consented. The time (mean +/- SD) from the index surgery to the follow-up visit was 4.0 +/- 0.5 years. MAIN OUTCOME MEASURES: Daily symptoms using a visual analogue scale for 30 consecutive days prior to clinical assessment; the amount of uterine bleeding was also assessed. Endometriosis recurrence was evaluated via clinical and ultrasonographic examination. RESULTS: The symptom sum scores (maximum 300) were low with median scores (range) of 3 (0-32) for dysmenorrhea and 9 (0-72) for pelvic pain. Evidence of RVE recurrence was found or suspected in 29 (48%) of the 60 women assessed. Clinical recurrence was not associated with pain symptoms. In univariable analysis, amenorrhea at the time of clinical assessment was associated with a lower risk of recurrence (odds ratio; OR 0.13; 95% CI (confidence interval) 0.02-0.65, p = 0.01); the effect of bowel resection was not significant (OR 0.37: 95% CI 0.13-1.07, p = 0.07). In multivariable analysis, the protective effect of bowel resection on recurrence was significant (OR 0.23; 95% CI 0.06-0.89, p = 0.03). CONCLUSIONS: Radical surgery may result in long-term pain relief in cases of RVE. Bowel resection is associated with a lower risk of RVE recurrence. Therapy that induces amenorrhea may be effective in preventing recurrence following surgical treatment of RVE. PMID- 20021268 TI - Evaluation of gonadotropin-releasing hormone agonist treatment for prevention of menstrual-related attacks in acute porphyria. AB - OBJECTIVE: To describe the benefits and adverse effects of gonadotropin-releasing hormone (GnRH) agonist treatment for prevention of recurrent menstrual attacks in women with acute intermittent porphyria and variegate porphyria. To describe concomitant add-back therapies with estradiol and progesterone and describe their benefits and adverse effects. DESIGN: A retrospective follow-up with questionnaires, interviews and medical records. SETTING: Out-patient care at the Umea University Hospital in Sweden. POPULATION: Sixteen Caucasian women with DNA diagnosed porphyria and menstrual-cycle-related porphyria attacks were treated with GnRH agonists during 1984-2000. Fourteen women participated. The mean age when treatment started was 33 years (17-48 years). The duration of treatment varied between 5 months and 9 years. METHODS: GnRH agonists were administered by the intranasal route or by injections. To reduce menopausal symptoms, add-back therapy with low doses of estradiol was administered, and for endometrial protection progesterone was usually administered. MAIN OUTCOME MEASURES: Treatment effects and adverse events as detected in questionnaires, interviews and medical records. RESULTS: Eleven women reported benefits from GnRH agonist treatment with less intense and/or less frequent porphyria attacks, and in four of them attacks almost disappeared. Two women reported no change. One woman had only temporary improvement. Porphyria attacks were triggered by solely estradiol add-back in two women and in five of nine women when progesterone was given. CONCLUSIONS: GnRH agonist treatment can ameliorate menstrual-cycle-related attacks of porphyria. Dose findings for GnRH agonists and add-back regimes especially for progesterone are intricate. PMID- 20021269 TI - Maternal serum placental growth factor and soluble fms-like tyrosine kinase 1 as early predictors of preeclampsia. AB - Abstract The aim of this study was to identify pregnant women at risk of preeclampsia (PE) before clinical manifestations appeared using a panel of serum markers. We recruited 240 consecutive women who presented for antenatal care. We investigated whether serum levels of placental growth factor (PlGF), its inhibitor, soluble fms-like tyrosine kinase-1 (sFlt-1), measured at 13-16 weeks gestation and the expression of fms-like tyrosine kinase-1 (Flt-1) in the maternal neutrophils measured by flow cytometry could be predictive of the subsequent development of PE. Serum PlGF levels were found to be significantly lower among women who developed PE than patients with gestational hypertension or patients in the control group (p < 0.001). In contrast, serum sFlt1 levels were most elevated in patients who developed PE versus those with gestational hypertension or the control group (p < 0.001). Serum levels of neutrophil-Flt-1, however, were lower in women who developed PE than in those with gestational hypertension or those in the control group (p < 0.001). Increased serum levels of sFlt-1, decreased levels of neutrophil-Flt-1, and decreased levels of PlGF may predict women at risk of developing PE later in pregnancy. PMID- 20021270 TI - Anchoring of vascular endothelial growth factor to surface-immobilized heparin on pancreatic islets: implications for stimulating islet angiogenesis. AB - In pancreatic islet transplantation, early revascularization is necessary for long-term graft function. We have shown in in vitro and in vivo models that modification with surface-attached heparin protects the islets from acute attack by the innate immune system of the blood following intraportal islet transplantation. In this study, we have investigated the ability of an immobilized conjugate composed of heparin to bind the angiogenic growth factor vascular endothelial growth factor-A (VEGF-A) as a means of attracting endothelial cells (ECs) to induce angiogenesis and revascularization. We analyzed the capacity of VEGF-A to bind to immobilized heparin and how this affected the proliferation and adherence of ECs to both artificial glass surfaces and islets. Quartz crystal microbalance with dissipation monitoring and slot-blot demonstrated the binding of VEGF-A to heparin-coated surfaces upon which ECs showed protein-dependent proliferation. Also, ECs cultured on heparin-coated glass surfaces exhibited effects upon focal contacts. Heparinized islets combined with VEGF-A demonstrated unaffected insulin release. Further, covering islets with heparin also increased the adhesion of ECs to the islet surface. Immobilized heparin on the islet surface may be a useful anchor molecule for achieving complete coverage of islets with angiogenic growth factors, ultimately improving islet revascularization and engraftment in pancreatic islet transplantation. PMID- 20021271 TI - Bioprocessing of human glioblastoma brain cancer tissue. AB - Solid cancer tumors are thought to arise from aberrant stem cell populations, called cancer stem cells (CSCs). Hence, the development of effective cancer therapies may rely on developing methods that specifically target these cells. However, the scarcity of CSCs in vivo represents a major impediment to such research, as there is an insufficient supply for basic biochemical and genetic analyses. It is therefore necessary to develop methods to expand reproducibly CSC tissue in vitro in a controlled environment. To date, we have developed bioreactor protocols for the suspension culture of an aggressive and deadly type of brain cancer called glioblastoma multiforme (GBM). Human GBM-derived cells achieved a maximum cell density of 2.4 x 10(6) cells/mL after 24 days under high shear conditions in batch culture conditions. In comparison, fed-batch cultures achieved 4.5 x 10(6) cells/mL after 32 days. Characterization of bioreactor expanded cells using both flow cytometry and a differentiation assay indicated that bioreactor-generated human GBM-derived cells have similar characteristics to the initial cell population and achieve >90% CD133 expression. Additionally, genomic characterization indicated that a very small number of key genes were differentially expressed in the bioreactor-expanded GBM-derived cells, thereby conserving the basic nature of the brain cancer tissue in the cell expansion process. PMID- 20021273 TI - Anti-inflammatory effects of lipoxins on lipopolysaccharide-induced uveitis in rats. AB - PURPOSE: Lipoxins exert potent anti-inflammatory and pro-resolving actions by reducing polymorphonuclear neutrophil (PMN) infiltration. This study describes the effect of lipoxin A4 and a stable analog on the resolution of ocular inflammation induced by intravitreal injection of lipopolysaccharides (LPS) in rats. METHODS: Six- to eight-week-old male Sprague Dawley (SD) rats were injected intravitreally with 2.5 microL physiologically balanced solution (LPS) containing 5 ng LPS, or 5 ng LPS + 50 ng LXA4 or 5 ng LPS + 50 ng 15-epi-LXA4 analog. Rats were anesthetized with intraperitoneal injection of a ketamine and xylazine cocktail. At 24 h, the animals were again anesthetized and the eyes examined for clinical signs of inflammation. The animals were then euthanized by CO2 inhalation and aqueous humor was collected in heparinized saline. Aqueous humor PMNs were counted using an Improved Neubauer Hemocytometer, and the protein concentration was determined by standard procedure. After enucleation, the eyes were dissected to remove the lens and the ocular tissues were frozen in liquid nitrogen and stored at -80 degrees C. Myeloperoxidase assay was done by a standard procedure. RESULTS: Compared to untreated LPS-injected controls, rats treated with either LXA4 or its stable analog had lower clinical inflammation score, significantly reduced aqueous humor PMN cell counts, aqueous humor protein levels, and the MPO values. The difference between the mean values of aqueous humor protein and MPO in the LXA4 and the analog injected eyes was not statistically significant, but PMN cell counts were significantly different. CONCLUSIONS: The ocular inflammatory response to intravitreally injected LPS in rats is significantly reduced by simultaneous injection of LXA4 or its analog. This finding supports an earlier independent observation of the ocular anti inflammatory effect of LXA4. Further investigation of lipoxins in the eye might offer a novel therapeutic approach to treating ocular inflammation in man. PMID- 20021274 TI - West Nile virus seroprevalence in blood donors from Central Anatolia, Turkey. AB - INTRODUCTION: West Nile virus (WNV) is a reemerging flavivirus that has displayed a drastic change in epidemiology in the last decade. Data on WNV activity in Turkey are currently limited. This study investigated WNV exposure in blood donors from Central Anatolia, Turkey. MATERIALS AND METHODS: A total of 2516 sera, collected from blood donors at four major branches of the Turkish Red Crescent Middle Anatolia Regional Blood Center, were evaluated by a commercial WNV immunoglobulin G (IgG) enzyme-linked immunosorbent assay (ELISA). Positive and borderline samples were investigated further by a WNV IgG indirect immunofluorescence test (IIFT), IgG ELISAs for tick-borne encephalitis virus and dengue virus, an IgG IIFT for yellow fever virus, and a multi-Flavivirus biochip IgG IIFT. WNV antibody specificity and titer values were determined by plaque reduction neutralization assay. IgG avidity and IgM were determined for confirmed samples. IgM-positive samples were also evaluated by a real-time reverse transcription polymerase chain reaction assay. RESULTS: Twenty-five samples (25/2516; 0.99%) were found reactive in the WNV ELISA/IIFT assays, and 14 could be confirmed by the plaque reduction neutralization assay (14/2516; 0.56%). All IgGs were of high avidity, and four samples (4/14; 28.6%), which were negative for viral RNA, were IgM positive. Although samples with neutralizing WNV IgGs had strong fluorescence intensity in IIFTs, no correlation between antibody titer values and IIFT intensity or quantitative ELISA results could be found. Three WNV nonreactive samples were positive in the dengue IgG ELISA test; one of these also displayed positive results for dengue virus in the mosaic biochip IIFT and reactivity in yellow fever virus IIFT. DISCUSSION: WNV exposure is confirmed in 0.56% of the tested healthy blood donors in Central Anatolia, with evidence for dengue/yellow fever and/or other flaviviral infections. This study is the first to document WNV exposure in individuals from Konya, Yozgat, and Sivas provinces in Central Anatolia, and it also establishes viral activity in Ankara, the capital of Turkey. PMID- 20021275 TI - Serological prevalence of hepatitis E virus in domestic animals and diversity of genotype 4 hepatitis E virus in China. AB - Pigs have been confirmed to be reservoirs of some genotypes of hepatitis E virus (HEV), and other nonhuman species are also likely infected with the virus. To assess the prevalence of HEV infection in domestic animals in China, 3579 serum samples, including 1967 swine, 700 goat, and 912 cattle sera, were collected from 26 provinces across the country and tested for HEV antibodies and antigen using enzyme immunoassays. The results showed that 82.2% of the swine samples, but only 10.4% and 28.2% of cattle and goat sera, were anti-HEV positive respectively. The prevalence of anti-HEV antibody in animals varied from province to province, ranging from 10.9% to 100% in pigs, 0% to 48% in goats, and 0% to 92.9% in cattle. About 1.9% of pigs, 1.6% of goats, and 0.8% of cattle tested in the study were positive for HEV antigen. Some samples, including all HEV antigen-positive samples, were tested for HEV-specific RNA using reverse transcription polymerase chain reaction. Fifteen swine samples, but none from the goats or cattle, were found to be HEV RNA positive. Sequence and phylogenetic analyses classified all the swine HEV isolates into HEV genotype 4, which was further divided into four subgroups. This study demonstrated that HEV infection is widespread in domestic animals, particularly pigs, in China. The HEV genotype infecting pigs in China was genotype 4. However, the isolates displayed considerable genetic diversity. PMID- 20021282 TI - Phospholipase A2 inhibitors as potential therapeutic agents for the treatment of inflammatory diseases. AB - IMPORTANCE OF THE FIELD: The various phospholipase A(2) (PLA(2)) types have been implicated in diverse kinds of lipid signaling and inflammatory diseases. Rheumatoid arthritis, lung inflammation, neurological disorders, such as multiple sclerosis, cardiovascular diseases, including atherosclerosis, and cancer are included among the diseases where PLA(2) enzymes are involved. Thus, there is a great interest in developing potent and selective PLA(2) inhibitors and some of them have entered clinical trials. AREAS COVERED IN THIS REVIEW: This review article discusses the role of each PLA(2) class in inflammatory diseases and the advances in the development of inhibitors presented in patent literature from January 2004 to May 2009. WHAT THE READER WILL GAIN: PLA(2)s cluster in four main types: secreted sPLA(2), cytosolic cPLA(2), Ca(2+)-independent iPLA(2) and lipoprotein-associated LpPLA(2). Each of those types has been implicated in diverse kinds of inflammatory diseases. Readers will rapidly gain an overview of the various PLA(2) inhibitors reported in the patent literature in the past 5 years. Furthermore, the readers will learn the difficulties related to the development of PLA(2) inhibitors as new drugs and also the different companies and research groups that are the main players in the field. TAKE HOME MESSAGE: Although the role of each PLA(2) is not yet distinct in different diseases, the development and future use of different PLA(2) inhibitors to treat human disease seems very promising. PMID- 20021283 TI - Inhibitors of diacylglycerol acyltransferase: a review of 2008 patents. AB - BACKGROUND: Postprandial hypertriglyceridemia has been identified as a major independent risk factor for future cardiovascular events. Therefore, inhibition of triglyceride synthesis has enormous therapeutic potential in the treatment of metabolic disorders. Diacylglycerol acyltransferase (DGAT) enzymes catalyze the final and only committed step in triglyceride biosynthesis and have thus been identified as potential therapeutic targets to combat human cardio-metabolic diseases. OBJECTIVE/METHOD: Significant interest in DGAT-1 inhibitors has emerged in the last several years. To provide a perspective on the exciting features of this enzyme for targeting metabolic diseases, a summary of the biology and pharmacology surrounding the DGAT enzymes is presented. Following this is a discussion of the various chemotypes that have been disclosed within relevant patent applications published in 2008. Specifically, the similarities and differences of the chemical structures and the biological data that are provided to support the corresponding claims are presented. CONCLUSION: Small molecule and biologic-based DGAT inhibitors have been successfully used for the preclinical validation of DGAT enzymes as targets for the treatment of metabolic diseases. Given the advanced stage in which some of the chemical matter resides, it is expected that DGAT inhibitors will enter the clinic in the coming years. PMID- 20021284 TI - Novel TACE inhibitors in drug discovery: a review of patented compounds. AB - TNF-alpha converting enzyme (TACE), a pro-inflammatory cytokine, catalyzes the formation of TNF-alpha from membrane bound TNF-alpha precursor protein. TNF-alpha is believed to play pathophysiological roles in inflammation, anorexia, cachexia, septic shock, viral replication and so on. TNF-alpha is a key player in inflammation and joint damage in rheumatoid arthritis. While a variety of TACE inhibitors have been reported in the literature, a vast majority of these compounds are peptidic and peptide-like compounds that are expected to have bioavailability and pharmacokinetic problems, common to such compounds, limiting their clinical effectiveness. Low molecular mass, long acting, orally bioavailable inhibitors of TACE are, therefore, highly desirable for the treatment of potential chronic diseases mentioned above. A review of patented compounds as TACE inhibitors in drug discovery is given. A selection of interesting patents recorded from 2001 to 2009 is presented. Various novel TACE inhibitors developed by different companies have been discussed. PMID- 20021285 TI - Picornaviral 3C protease inhibitors and the dual 3C protease/coronaviral 3C-like protease inhibitors. AB - IMPORTANCE OF THE FIELD: Picornaviruses are small non-enveloped RNA viruses with genomic RNA of 7500 - 8000 nucleotides, whereas coronaviruses (CoV) are RNA viruses with larger genome of 27 - 32 kb. Both types of viruses translate their genetic information into polyprotein precursors that are processed by virally encoded 3C proteases (3C(pro)) and 3C-like proteases (3CL(pro)), respectively, to generate functional viral proteins. The most studied human rhinoviruses (HRV) belonging to picornaviridae family are the main etiologic agents of the common cold. Due to lack of effective drugs, 3C(pro) has served as an excellent target for anti-viral intervention and considerable efforts have been made in the development of inhibitors. Interestingly, the inhibitors of 3C(pro) cannot inhibit 3CL(pro) potently without modification due to subtle differences in their active-site structures, but a group of common inhibitors against 3C(pro) and 3CL(pro) were found recently. AREAS COVERED IN THIS REVIEW: The inhibitors against 3C(pro) reported in the literatures and patents, with a focus on those inhibiting HRV and the dual picornaviral 3C(pro)/coronaviral 3CL(pro) inhibitors, are summarized in this review. WHAT THE READERS WILL GAIN: Readers will rapidly gain an overview of the individual and dual 3C(pro) inhibitors and the structural basis for discriminating them. TAKE HOME MESSAGE: In the future, more selective potent inhibitors against each protease and dual inhibitors against both proteases can be further developed to treat the diseases caused by picornaviruses and CoV. PMID- 20021286 TI - Radiation countermeasure agents: an update. AB - IMPORTANCE OF THE FIELD: Ionizing radiation (IR) can produce deleterious effects in living tissues, leading to significant morbidity and a potentially fatal illness affecting various organs dose-dependently. As people may be exposed to IR during cancer radiotherapy or as a result of a radiological/nuclear incident or act of terrorism, the danger of irradiation represents a serious public health problem. At present, however, this problem remains largely impervious to medical management. There is, therefore, a pressing need to develop safe and effective radiation countermeasure (RC) agents to prevent, mitigate or treat the harmful consequences of IR exposure. AREAS COVERED IN THIS REVIEW: Recent advances in the search for RC agents as reflected by the relevant patent literature of the past five years along with peer-reviewed publications are surveyed. WHAT THE READER WILL GAIN: A total of 43 patents, describing approximately 38 chemically diverse compounds with RC potential are analyzed. These include antioxidants capable of scavenging IR-induced free radicals, modulators of cell death signaling or cell cycle progression, cytokines or growth factors promoting tissue repair and inhibitors of inflammatory cytokines. TAKE HOME MESSAGE: Several of these RC candidates appear promising, including at least two that are undergoing evaluation for fast-track clinical development. PMID- 20021287 TI - Anti-VEGF agents for age-related macular degeneration. AB - Age-related macular degeneration (AMD) is a leading cause of legal blindness in the elderly in the industrialized world and the third major cause of blindness around the globe. Although neovascular AMD is less prevalent than atrophic AMD, it accounts for most cases with severe visual loss from AMD. VEGF seems to be a key contributary factor in the pathophysiology underlying neovascular AMD. Until recently, treatment options for neovascular AMD were limited. With the recent development of anti-VEGF therapies that have demonstrated efficacy in studies with broad eligibility criteria, the repertoire of treatments for neovascular AMD has been significantly expanded to now include the various recognized angiographic lesion subtypes. To discuss recent anti-VEGF agents in the management of AMD. Although therapy with anti-VEGF agents is the gold standard with promising results, many intravitreal injections are often required, and they do not cure all cases of wet AMD. With the recent advances in the medical therapy of exudative AMD, there is reason to be optimistic about future management of AMD as well. PMID- 20021289 TI - Heterocyclic indazole derivatives as SGK1 inhibitors, WO2008138448. AB - Heterocyclic indazole derivatives are claimed in patent WO2008138448 as inhibitors of the serum- and glucocorticoid-inducible-kinase 1 (SGK1) and drugs for the pharmacological treatment of SGK1-related diseases, such as diabetes, obesity, metabolic syndrome, systemic and pulmonary hypertension, cardiac fibrosis, hypertrophy and insufficiency, arteriosclerosis, glomerulosclerosis, nephrosclerosis, nephritis, nephropathy, deranged electrolyte excretion, fibrosing and inflammatory disease (e.g., liver cirrhosis, lung fibrosis, rheumatism, arthrosis, Crohn s disease, chronic bronchitis, radiation fibrosis, sclerodermia, cystic fibrosis, scar formation and Alzheimer' disease), tumor growth, peptic ulcers and some disorders hitherto not conclusively shown to involve SGK1. Most of the claims are supported by the literature. SGK1 is ubiquitously expressed and its expression is stimulated by hyperglycemia, cell shrinkage, ischemia, glucocorticoids, mineralocorticoids and several inflammatory mediators including TGF-ss. SGK1 is activated by insulin and growth factors via the phosphatidylinositol-3-kinase pathway. SGK1 regulates ion channels (including ENaC, KCNE1/KCNQ1), carriers (including NCC, NHE3, SGLT1), Na(+)/K(+)-ATPase, enzymes (including glycogen-synthase-kinase-3) and transcription factors (including FOXO3a, ss-catenin, NF-kappaB). A gain-of-function SGK1 gene variant, carried by approximately 3 - 5% of Caucasians and approximately 10% of Africans, is associated with increased blood pressure, obesity and type 2 diabetes. In vitro and in vivo experiments suggested a critical role of SGK1 in renal fluid retention and hypertension, glucose-induced obesity, coagulation and increased matrix protein formation. PMID- 20021290 TI - Clinical outcomes and nephrotoxicity associated with vancomycin trough concentrations during treatment of deep-seated infections. AB - OBJECTIVE: Higher vancomycin concentrations are thought necessary for treatment of deep-seated methicillin-resistant Staphylococcus aureus (MRSA) infection, yet this may result in greater risk of nephrotoxicity. We evaluated the relationship of serum vancomycin trough concentration with clinical outcomes and nephrotoxicity for patients with deep-seated MRSA infection. METHODS: A retrospective cohort study evaluated adults with MRSA pneumonia, endocarditis or osteomyelitis who received vancomycin for > or = 5 days from June 2005 to June 2007. Patients were stratified by mean vancomycin trough level [low (< 15 mg/l), high (> or = 15 mg/l)]. Outcomes were clinical response, mortality, length of stay (LOS) and nephrotoxicity. Three definitions of nephrotoxicity were used: i) rise in serum creatinine (SCr) > or = 0.5 mg/dl; ii) 50% increase in SCr; and iii) 25% decrease in estimated creatinine clearance. RESULTS: Fifty-five patients experiencing MRSA pneumonia (n = 28), endocarditis (n = 9), osteomyelitis (n = 20) and multiple infections (n = 2) were stratified into low (n = 39) and high (n = 16) groups. High group patients were more likely to be septic (p = 0.01) and have a higher APACHE II score (p = 0.01). After adjustment for APACHE II score, clinical response rates among survivors did not differ significantly. Risk of death was not significantly different between the high (19%) and low (5%) group patients (p = 0.1). LOS did not differ significantly between groups (p = 0.7). Nephrotoxicity occurred in the low and high groups, respectively, for 10 and 31% (p = 0.04) with definition 1, 10 and 31% (p = 0.04) with definition 2, and 13 and 25% (p = 0.1) with definition 3. After adjustment for APACHE II score, odds of nephrotoxicity based on definitions 1 or 2 were increased among the high versus low groups (OR = 3.27, 95% CI: 0.7 - 15.25, p = 0.1), although not statistically significant. CONCLUSIONS: Clinical outcomes did not differ significantly between high and low trough groups for deep-seated MRSA infections. Nephrotoxicity was consistently higher in the high trough group, regardless of the definition used. PMID- 20021291 TI - Genetic polymorphism of inosine-triphosphate-pyrophosphatase influences mercaptopurine metabolism and toxicity during treatment of acute lymphoblastic leukemia individualized for thiopurine-S-methyl-transferase status. AB - IMPORTANCE OF THE FIELD: Although genetic polymorphisms in the gene encoding human thiopurine methyltransferase (TPMT) are known to have a marked effect on mercaptopurine metabolism and toxicity, there are many patients with wild-type TPMT who develop toxicity. Furthermore, when mercaptopurine dosages are adjusted in patients who are heterozygous at the TPMT locus, there are still some patients who develop toxicity for reasons that are not fully understood. Therefore, we recently studied the effects of a common polymorphism in another gene encoding an enzyme involved in mercaptopurine metabolism (SNP rs1127354 in inosine triphospate-pyrophosphatase, ITPA), showing that genetic polymorphism of ITPA is a significant determinant of mercaptopurine metabolism and of febrile neutropenia following combination chemotherapy of acute lymphoblastic leukemia (ALL) in which mercaptopurine doses are individualized based on TPMT genotype. AREA COVERED IN THIS REVIEW: In this review, we summarize the knowledge available about the effect and clinical relevance of TPMT and ITPA on mercaptopurine pharmacogenomics, with a particular focus on the use of this medication in pediatric patients with ALL. WHAT THE READER WILL GAIN: Reader will gain insights into: i) the effects of pharmacogenomic traits on mercaptopurine toxicity and efficacy for the treatment of ALL and ii) individualization strategies that can be used to mitigate toxicity without compromising efficacy in pediatric patients with ALL. TAKE HOME MESSAGE: Mercaptopurine dose can be adjusted on the basis of TPMT genotype to mitigate toxicity in pediatric patients with ALL. As treatment is individualized in this way for the most relevant genetic determinant of drug response (i.e., for mercaptopurine, TPMT), the importance of other genetic polymorphisms emerges (e.g., ITPA). PMID- 20021292 TI - Clinically based evidence of drug-herb interactions: a systematic review. AB - IMPORTANCE OF THE FIELD: Healthcare practitioners are deeply concerned about drug herb interactions and how concurrent administration may affect both the safety and effectiveness of prescribed drugs. Interactions between botanical medicines and synthetic drugs can be clinically relevant and it is important to understand what kinds of interactions are possible. Better knowledge in this area will help avoid negative interactions and may also help enable synergistic interactions. AREAS COVERED IN THIS REVIEW: Includes articles related to the investigation of Western botanicals or whole herbal extracts in human subjects, investigating either the impact on Cytochrome P450 isoenzymes or an assessment of specific drug herb interactions within a clinical trial. Searches were conducted in both Pubmed and EMBASE from inception to March 2009. WHAT THE READER WILL GAIN: Knowledge regarding specific interactions to consider in clinical practice. The reader will also gain an appreciation of the complexities associated with the area of drug herb interactions. Summary tables of relevant drug-herb interactions are presented both for the individual herbs included and at the level of the CYP450 enzymes. TAKE HOME MESSAGE: Knowledge of drug-herb interactions is limited and much more research needs to be done to further document clinically relevant interactions. Even though preclinical data are often poorly generalizable to the human situation, caution must be taken in the absence of clinical evidence especially where drugs with narrow therapeutic windows are concerned. PMID- 20021293 TI - Voriconazole therapeutic drug monitoring: focus on safety. AB - IMPORTANCE OF THE FIELD: Voriconazole has been widely used for the treatment of invasive fungal diseases, particularly invasive aspergillosis. Drug-drug interactions are, however, the main drawback associated with voriconazole use, since this drug suffers from extensive hepatic metabolism. AREAS COVERED IN THIS REVIEW: This article reviews the current literature on voriconazole therapeutic drug monitoring, with a special focus on drug safety. WHAT THE READER WILL GAIN: An update on voriconazole metabolism, drug interactions, toxicity and the relation of these with voriconazole drug concentrations. TAKE HOME MESSAGE: Therapy with voriconazole may be better guided by measuring voriconazole concentrations in the plasma. PMID- 20021294 TI - Safety and tolerability of transdermal and oral rivastigmine in Alzheimer's disease and Parkinson's disease dementia. AB - IMPORTANCE OF THE FIELD: Cholinesterase inhibitors are the mainstay of symptomatic therapy for Alzheimer's disease (AD). Rivastigmine, an inhibitor of both acetylcholinesterase and butyrylcholinesterase, is available as a transdermal patch and in oral forms. It is also approved for the treatment of Parkinson's disease dementia (PDD) in many countries. The objective of this article is to review the safety and tolerability profile of transdermal and oral rivastigmine in AD and PDD patients. AREAS COVERED IN THIS REVIEW: Articles were identified by searching MEDLINE in July 2009 using the terms rivastigmine, Exelon, ENA 713 and clinical trial. All double-blind, placebo-controlled randomized trials in which rivastigmine monotherapy was administered to AD or PDD patients for longer than 2 weeks were included. WHAT THE READER WILL GAIN: This article provides a comprehensive summary of currently available safety data on rivastigmine. TAKE HOME MESSAGE: The main adverse events reported with rivastigmine therapy are gastrointestinal in nature. However, the transdermal patch appears to reduce these side effects, allowing more patients to access higher therapeutic doses. Overall, the safety profile of rivastigmine is favorable and the improved tolerability offered by the rivastigmine patch suggests that transdermal delivery may be the best way to deliver this drug in AD and PDD patients. PMID- 20021295 TI - The neurobiology, clinical efficacy and safety of acamprosate in the treatment of alcohol dependence. AB - IMPORTANCE TO THE FIELD: Acamprosate, marketed under the brand name Campral, (Forest Pharmaceuticals, Inc., Saint Louis, MO, USA; Merck Sante s.a.s., Lyon, France) is an orally administered drug approved in the US and throughout much of the world for treating alcohol dependence. Its safety and efficacy have been demonstrated in a number of clinical trials worldwide and as with all pharmacotherapies for alcoholism, it is used in conjunction with psychosocial interventions. AREAS COVERED IN THIS REVIEW: This article reviews the mechanism of action, clinical efficacy and safety of acamprosate in Phase I, II and III randomized controlled trials involving healthy and alcohol-dependent populations using published reports from 1984 to 2009. WHAT THE READER WILL GAIN: This review provides an update of the mechanism of action and the safety and efficacy profile of acamprosate. TAKE HOME MESSAGE: Acamprosate appears to act centrally to restore the normal activity of glutamatergic neurotransmission altered by chronic alcohol exposure. Acamprosate's excellent safety profile along with several pharmacokinetic and pharmacodynamic characteristics make it well suited for treating a broad population of alcohol-dependent patients. PMID- 20021296 TI - Gonadal dysfunction and pelvic sonographic findings in females with thalassaemia major. AB - OBJECTIVE: To investigate pubertal and menstrual problems and evaluate pelvic sonographic findings in patients with beta-thalassaemia major. MATERIAL AND METHODS: Twenty-five female patients followed for thalassaemia major constituted the study population. Sexual maturation and hormonal status were assessed. Pubertal and menstrual problems were investigated. RESULTS: There was one patient with delayed puberty and five patients with arrested puberty. Mean ferritin level in this group of patients was slightly but not significantly higher than patients with normal pubertal maturation (2620 +/- 994 ng/ml vs. 2409 +/- 1348 ng/ml, p > 0.05). There were 10 patients with primary amenorrhoea, three with secondary amenorrhoea, five with oligomenorrhoea and irregular menstruation and one with regular menstruation. Compared to menstruating patients, the mean uterine size was smaller (4.1 +/- 3.5 cm(3) vs. 52.8 +/- 14.5 cm(3)) in all patients with delayed and arrested puberty (p < 0.05). Ten patients were taking hormone replacement therapy (HRT). The mean uterine size in these patients was larger than that in patients with amenorrhoea who were not taking HRT, but smaller than that in menstruating patients (9.1 +/- 15.9 cm(3), 2.7 +/- 1.3 cm(3) and 52.8 +/- 14.5 cm(3), respectively) (p < 0.05). CONCLUSION: Thalassaemia major has important side effects on the hypothaloma-pituitary-gonadal axis resulting in pubertal and menstrual abnormalities. HRT should be given to provide normal sexual maturation in these patients. PMID- 20021297 TI - Dental caries vaccines: prospects and concerns. PMID- 20021298 TI - Filovirus vaccines: what challenges are left? PMID- 20021300 TI - 3rd Global Vaccine Congress. AB - Singapore was the location of the 3rd Global Vaccine Congress on 4-6 October 2009, and it provided a suitable place for sharing a range of developments, concepts and challenges associated with vaccines. One of the major goals of this meeting was to present a broad and balanced program of research, development, production, clinical testing and delivery of vaccines to people living in different parts of the world. This report contains an overview of recent developments in the vaccine field against a range of infections and disease situations that were presented during the meeting. The highlights included challenges to the current constraints on the timely and cost-effective development of vaccines and a focus on the H1N1 influenza pandemic concerns. The meeting also generated presentations that challenged the regulatory and political status quo in order to expand global access to vaccines and facilitate speedier and more cost-effective development of new vaccines. PMID- 20021301 TI - Pathogen sensing in innate immunity. AB - TLRs (Toll-like receptors), NLRs (Nod-like proteins) and RLRs (RIG-like receptors), pathogens sensors of innate immunity conference organized by EuroSciCon hosted a large group of scientists from across Europe. The Chairs hoped the meeting would "provide an overview of these three families of receptors and provide the most recent advances in the area of innate immune pattern recognition", with a view to informing future directions in this field. Eleven talks and six posters were presented; the Chairs gave a detailed introduction, followed by a detailed summation at the end. This article aims to describe the key topics highlighted. PMID- 20021302 TI - Topical resiquimod: a promising adjuvant for vaccine development? AB - The development of vaccines for cancer and infectious diseases that require the predominant induction of cell-mediated immunity, including effective cytotoxic T lymphocyte (CTL) responses, remains a challenge. Over the past few years, numerous studies have demonstrated that targeting of antigens to immune receptors expressed by dendritic cells (DCs), such as Toll-like receptors (TLRs), elicits potent CTL responses. Therefore, the design of new adjuvants has focused on TLR agonists that elicit maturation of DCs to effectively link the innate and adaptative immune responses. The paper under evaluation describes the topical use of resiquimod, a TLR7/8 agonist, to enhance cross-priming to subcutaneously administered ovalbumin. The results obtained demonstrated that topical resiquimod is a potent adjuvant for subcutaneous vaccines, inducing clinically relevant antigen-specific CTL responses, which protect against B16-ovalbumin tumor challenge in mice. PMID- 20021303 TI - It's all in for the HER family in tumorigenesis. AB - The EGF receptor family is a group of receptor tyrosine kinases that have been implicated in the development of a variety of malignancies. As such, they have been targeted in the generation of pharmacologic agents, several of which have been approved as anti-tumor therapeutics. The lone exception is ERBB4, for which the function and relationship to cancer are not yet clear and no targeted therapies exist. The paper under evaluation demonstrates a role for ERBB4 mutations in the development of melanoma. It identifies ERBB4 mutations present in melanomas that augment proliferation and cell survival and thus contribute to dysregulated growth. Furthermore, it shows that agents targeting the EGF receptor family can reduce the proliferation of melanoma cells harboring these mutations. These findings further emphasize the role of the ERBB subfamily in tumorigenesis and establish ERBB4 as a new target in the development of anti-tumor strategies. PMID- 20021304 TI - Perspectives for the development of human papillomavirus vaccines and immunotherapy. AB - Human papillomavirus (HPV) is the most common sexually transmitted infection and is responsible for 90-99% of cervical cancer (CxCa) cases. Although effective screening programs have reduced the incidence of CxCa in developed countries, they are often not well organized. Prophylactic vaccination against HPV seems to be a good strategy for the prevention of CxCa. However, because millions of women are already infected with HPV, therapeutic HPV vaccines need to be developed further to treat these women. This review discusses the actual perspectives on both HPV vaccines and immunotherapy worldwide. In addition, some of the perspectives in France are also briefly discussed. PMID- 20021305 TI - Prophylactic HPV vaccines: the Finnish perspective. AB - Finland is a country with a dual role in the human papillomavirus (HPV) field. Finnish scientists have pioneered in HPV research and participated in international HPV vaccine trials, but officially Finland is reluctant to implement HPV vaccines into its national vaccination program owing to the reasons discussed in this article. In contrast to other European countries, Finnish authorities were reluctant to initiate the evaluation process for HPV vaccines after licensure. Instead of prompt implementation, it was decided that a long term prospective (Phase IV effectiveness) study to evaluate the bivalent HPV vaccine in a randomized community trial should be started. In addition, the Finnish authorities refuse to accept the compelling scientific evidence on the efficacy and safety of HPV vaccines. They report that cervical cancer (CC) is effectively controlled by the national screening program, despite the fact that CC in Finland has increased very rapidly since 1992. The Finnish perspective on HPV vaccines appears to postpone all critical decisions until the second half of the 2010s. This will have two direct consequences: 1) every year of delay means that an entire birth cohort (>60,000 girls and boys) will lose the opportunity of being offered the state-of-art cancer prevention provided by prophylactic HPV vaccines; and 2) given the declining trends of CC elsewhere, within a few years Finland will fall among the countries with a high incidence of CC. It is the personal conviction of this author that there are no sustainable arguments remaining that advocate continuing this Finnish perspective on HPV vaccines any further. PMID- 20021306 TI - Animal models for the preclinical evaluation of candidate influenza vaccines. AB - At present, new influenza A (H1N1)2009 viruses of swine origin are responsible for the first influenza pandemic of the 21st Century. In addition, highly pathogenic avian influenza A/H5N1 viruses continue to cause outbreaks in poultry and, after zoonotic transmission, cause an ever-increasing number of human cases, of which 59% have a fatal clinical outcome. It is also feared that these viruses adapt to replication in humans and become transmissible from human to human. The development of effective vaccines against epidemic and (potentially) pandemic viruses is therefore considered a priority. In this review, we discuss animal models that are used for the preclinical evaluation of novel candidate influenza vaccines. In most cases, a tier of multiple animal models is used before the evaluation of vaccine candidates in clinical trials is considered. Commonly, vaccines are tested for safety and efficacy in mice, ferrets and/or macaques. The use of each of these species has its advantages and limitations, which are addressed here. PMID- 20021307 TI - Strategies for differentiating infection in vaccinated animals (DIVA) for foot and-mouth disease, classical swine fever and avian influenza. AB - The prophylactic use of vaccines against exotic viral infections in production animals is undertaken exclusively in regions where the disease concerned is endemic. In such areas, the infection pressure is very high and so, to assure optimal protection, the most efficient vaccines are used. However, in areas considered to be free from these diseases and in which there is the possibility of only limited outbreaks, the use of Differentiation of Infected from Vaccinated Animals (DIVA) or marker vaccines allows for vaccination while still retaining the possibility of serological surveillance for the presence of infection. This literature review describes the current knowledge on the use of DIVA diagnostic strategies for three important transboundary animal diseases: foot-and-mouth disease in cloven-hoofed animals, classical swine fever in pigs and avian influenza in poultry. PMID- 20021308 TI - Mechanisms of T-cell inhibition: implications for cancer immunotherapy. AB - Cancer vaccines designed to augment effector T-cell responses have been disappointing with respect to clinical efficacy. This lack of effectiveness may be due to the fact that regulatory mechanisms, both intrinsic and extrinsic to activated T cells, play important roles in inhibiting vaccine-induced effector T cell responses. This concept raises the possibility that blockade of these regulatory checkpoints might enhance anti-tumor immune responses. In this review, we discuss several regulatory mechanisms that act to control effector T-cell responses and identify strategies to circumvent these mechanisms in order to improve clinical responses. PMID- 20021311 TI - Leptin, visfatin, insulin resistance, and body composition change in chronic obstructive pulmonary disease. AB - OBJECTIVE: The aim of the study was to compare endocrine parameters such as leptin, visfatin, insulin resistance, exercise capacity and body composition change, the pulmonary functions test (PFT) and arterial blood gases (ABG) parameters of chronic obstructive pulmonary disease (COPD) patients and in healthy controls. MATERIALS AND METHOD: Fifty-five patients with COPD and without malnutrition and 25 healthy controls were included in our study. The serum leptin, visfatin, tumor necrosis factor alpha (TNF-alpha) and insulin resistance, body fat-free mass (FFM) and fat mass (FM) were measured in the groups. Additionally, body mass index (BMI) was calculated and the 6-minute walk test (6MWT), PFT and ABG analyses were performed in all of the cases. RESULTS: No difference in BMI between the COPD group and controls was determined. Serum leptin and visfatin levels, FFM and 6MWT distance were significantly lower in the patients with COPD (p < 0.001, p = 0.001, p = 0.032, p < 0.001, respectively). A correlation was found between serum leptin levels and BMI (r = 0.333, p = 0.027), and with FM (r = 0.365, p = 0.029). Serum visfatin level was correlated with the percentage of forced expiratory volume in the first second in the patients with COPD (r = 0.371, p = 0.013). HOMA-IR (Homeostasis model assessment of insulin resistance) and serum TNF-alpha levels in the patients with COPD were found to be significantly higher than controls (p = 0.001, p < 0.001). CONCLUSION: These results may be earlier signs for further diseases that can emerge in the advanced stages in patients with COPD. Evaluating the patients not only with the pulmonary function and also systemically, contributes to minimizing the mortality and morbidity. PMID- 20021312 TI - Could carbon monoxide and bilirubin be friends as well as foes of the body? AB - Endogenous carbon monoxide (CO) production was first described 60 years ago. CO is a by-product of the metabolism of haeme to biliverdin. This, in turn, becomes bilirubin. During the past 15 years epidemiological studies and animal experiments have identified bilirubin as a molecule at the crossroads of the protection of the body against reactive oxygen species (ROS). The studies have focused on bilirubin as a biomarker of arterial disease. Recently the potential of CO as a therapeutic agent has been explored. This review assesses the current state of evidence and sets the data in the context of whether CO is an endogenous signalling molecule, a marker of vascular disease and, whether, together with bilirubin, CO could be a potential therapeutic agent. PMID- 20021313 TI - Impaired glucose tolerance and endothelial damage, as assessed by levels of von Willebrand factor and circulating endothelial cells, following acute myocardial infarction. AB - BACKGROUND: Impaired glucose tolerance (IGT) following acute myocardial infarction (AMI) increases the incidence of major adverse cardiac events. We hypothesized that endothelial damage following AMI, as assessed by levels of von Willebrand factor (vWF) and circulating endothelial cells (CECs), would be more pronounced in patients with IGT compared to those with normal glucose tolerance (NGT). METHODS: We studied non-diabetic patients with AMI (n=125; 107 (86%) male; mean age 59 years (SD 12.5)) who underwent oral glucose tolerance testing 3-5 days after admission. We measured vWF (enzyme-linked immunosorbent assay) and CECs (CD146 immunobead capture) in the fasting state and at 2 h post glucose load. RESULTS: Base-line vWF and CEC levels were higher in IGT patients versus those with NGT and healthy controls (HC) (P<0.001). The acute increase in vWF and CECs in response to the glucose load was significantly higher in the IGT group compared to those with NGT and HC (P<0.01)-an increase on a par with that seen in newly diagnosed diabetics. CONCLUSION: The degree of endothelial damage post AMI in patients with IGT is greater than NGT, and comparable to that seen in frank diabetes mellitus. Subjects with IGT therefore need to be as actively sought and managed. PMID- 20021314 TI - Where will insights into hippocampal activity in schizophrenia lead us? PMID- 20021315 TI - A model of suicidal behavior among depressed immigrants. PMID- 20021317 TI - Epilepsy and bipolar disorders. AB - Although mood disorders represent a frequent psychiatric comorbidity among patients with epilepsy, data regarding bipolar disorders are still limited. However, these two conditions apparently share a number of biochemical and pathophysiological underpinnings, such as the kindling phenomenon, changes in neurotransmitters and modifications in voltage-opened ion channels and second messenger systems. Moreover, epilepsy and bipolar disorders are both episodic conditions with a time course of illness that can become chronic. Recent data suggest that mood instability is actually frequent among patients with epilepsy but is phenomenologically different from that described in bipolar disorder. The present paper reviews available evidence regarding such an intriguing comorbidity in order to discuss valuable clinical issues and to suggest novel therapeutic strategies. PMID- 20021318 TI - The role of copy number variation in schizophrenia. AB - Recent developments in microarray technology have revealed the presence of many submicroscopic deletions and duplications in the human genome. Some of these have been found to increase the risk for neuropsychiatric disorders. Over the last 2 years, several large studies on schizophrenia have implicated large deletions and duplications that increase the risk of developing this disorder. It is now clear that rare deletions at 1q21.1, 15q13.3, 15q11.2 and 22q11.2, as well as duplications at 16p11.2 and 16p13.1, increase the risk of developing schizophrenia. They are found collectively in up to 3% of patients; therefore, they account for only a small proportion of the genetic causes of schizophrenia. In this paper I will review the evidence for these findings. PMID- 20021319 TI - Progressive structural brain changes in schizophrenia. AB - Several important progressive brain changes occur in schizophrenia. Continuous progressive brain tissue decreases and lateral ventricular volume increases in chronically ill patients, up to at least 20 years after their first symptoms. The total duration of psychosis may be negatively associated with the grey matter volume change, and positively associated with the volume change of each of the lateral ventricles and the third ventricle. There are progressive frontal changes in males with adolescent-onset psychosis. Ultra high-risk patients who subsequently develop psychosis and first-episode psychosis patients develop significant grey matter reduction in the planum polare, planum temporale and caudal region; a progressive process in the superior temporal gyrus may precede the first expression of florid psychosis. The rate of decline in the sizes of the putamen and corpus callosum is greater in patients with a poor outcome compared to those with a good-outcome. In addition to describing these phenomena, their potential causes are discussed. PMID- 20021321 TI - The mononuclear phagocyte system and its cytokine inflammatory networks in schizophrenia and bipolar disorder. AB - This review describes patients with schizophrenia and bipolar disorder. In such patients, a high inflammatory set point of circulating monocytes at the transcriptome level is observed, involving various inflammatory transcripts forming distinct fingerprints (the transcriptomic monocyte fingerprint in schizophrenia overlaps with that in bipolar disorder, but also differs with it at points). There are increased levels of compounds of the IL-1, IL-6 and TNF system in the serum (be it modest and inconsistent). There is also evidence that the IL 2 system is activated in patients with schizophrenia (and perhaps those with mania), although independently of the activation of the IL-1, IL-6 and TNF systems, suggesting separate inducing mechanisms for monocyte and T-cell activation. It is not yet known whether such T cell activation involves the Th1/Th2/Th17 or Treg systems. PMID- 20021320 TI - Effect of second-generation antipsychotics on cognition: current issues and future challenges. AB - Generalized cognitive impairments are stable deficits linked to schizophrenia and key factors associated with functional disability in the disorder. Preclinical data suggest that second-generation antipsychotics could potentially reduce cognitive impairments; however, recent large clinical trials indicate only modest cognitive benefits relative to first-generation antipsychotics. This might reflect a limited drug effect in humans, a differential drug effect due to brain alterations associated with schizophrenia, or limited sensitivity of the neuropsychological tests for evaluating cognitive outcomes. New adjunctive procognitive drugs may be needed to achieve robust cognitive and functional improvement. Drug discovery may benefit from greater utilization of translational neurocognitive biomarkers to bridge preclinical and clinical proof-of-concept studies, to optimize assay sensitivity, enhance cost efficiency, and speed progress in drug development. PMID- 20021322 TI - Therapy-resistant depression. AB - Therapy-resistant depression is not untreatable. Numerous treatment methods with substantiated effectiveness exist. However, none of them are able to ensure remission. Thus, what matters when treating therapy-resistant depression is the strict, step-by-step deployment of existing therapeutic options at the right tempo and with a regular systematic evaluation of patient response. Remission should always be the aim, not just some abatement in symptoms (response). The following should be avoided: too small dosages of medication; changing the therapeutic strategy too frequently or too quickly; thoughtlessly sticking to an ineffective treatment over too long a period of time; or unsystematic poly pharmacy. If these are all avoided than therapy-resistant depressive patients will have a good chance of recovery. PMID- 20021323 TI - Efficacy of vagus nerve stimulation in the treatment of depression. AB - Major depressive disorder is a disease with prominent individual, medical and economic impacts. A relevant proportion of depressive patients suffering from a therapy-resistant course are increasingly being treated with antidepressant brain stimulation techniques. One of these interventions is the vagus nerve stimulation that has recently been tested in a number of clinical trials. To date, the acute and long-term efficacy of vagus nerve stimulation are still under debate. Thus further studies are required, especially since the exact mode of action of vagus nerve stimulation is still not well understood. In this paper we will review the results of existing clinical trials as well as the neurobiological effects measured with neuroimaging, biochemical and electrophysiology approaches. PMID- 20021325 TI - Sleep and headache: a bidirectional relationship. AB - Sleep and pain perception are two phylogenetically well-conserved functions, strictly influenced by environmental and psychological factors, and are able to interact reciprocally both in physiological and pathological situations. Sleep and head-pain perception share the involvement of several structures, such as the thalamus, the hypothalamus and brainstem nuclei, including the locus coeruleus and raphe nuclei. There ais clinical evidence indicating that sleep disorders can precede the appearance of certain headaches and that head pain, especially when frequent, can, in turn, affect sleep quality. In the present work the anatomy, physiology and pathology of sleep and head-pain perception will be reviewed with the aim of highlighting the points of contact and possible unifying treatment strategies. PMID- 20021326 TI - Drug interactions involving the new second- and third-generation antiepileptic drugs. AB - During the period 1989-2009, 14 new antiepileptic drugs (AEDs) were licensed for clinical use and these can be subdivided into new second- and third-generation AEDs. The second-generation AEDs comprise felbamate, gabapentin, lamotrigine, levetiracetam, oxcarbazepine, pregabalin, rufinamide, stiripentol, tiagabine, topiramate, vigabatrin and zonisamide. The third-generation AEDs comprise eslicarbazepine acetate and lacosamide. The interaction propensity of AEDs is very important because all new AEDs are licensed, at least in the first instance, as adjunctive therapy. The present review summarizes the interactions (pharmacokinetic and pharmacodynamic) that have been reported with the newer AEDs. The pharmacokinetic interactions include those relating to protein-binding displacement from albumin in blood, and metabolic inhibitory and induction interactions occurring in the liver. Overall, the newer AEDs are less interacting because their pharmacokinetics are more favorable and many are minimally or not bound to blood albumin (e.g., eslicarbazepine, felbamate, gabapentin, lacosamide levetiracetam, rufinamide, topiramate and vigabatrin) and are primarily renally excreted or metabolized by noncytochrome P450 or uridine glucoronyl transferases (e.g., gabapentin, lacosamide levetiracetam, rufinamide, topiramate and vigabatrin) as opposed to hepatic metabolism which is particularly amenable to interference. Gabapentin, lacosamide, levetiracetam, pregabalin and vigabatrin are essentially not associated with clinically significant pharmacokinetic interactions. Of the new AEDs, lamotrigine and topiramate are the most interacting. Furthermore, the metabolism of lamotrigine is susceptible to both enzyme inhibition and enzyme induction. While the metabolism of felbamate, tiagabine, topiramate and zonisamide can be induced by enzyme-inducing AEDs, they are less vulnerable to inhibition by valproate. Noteworthy is the fact that only five new AEDs (eslicarbazepine, felbamate, oxcarbazepine, rufinamide and topiramate) interact with oral contraceptives and compromise contraception control. The most clinically significant pharmacodynamic interaction is that relating to the synergism of valproate and lamotrigine for complex partial seizures. Compared with the first-generation AEDs, the new second- and third generation AEDs are less interacting, and this is a desirable development because it allows ease of prescribing by the physician and less complicated therapeutic outcomes and complications for patients. PMID- 20021324 TI - Temporal characteristics of botulinum neurotoxin therapy. AB - Botulinum neurotoxin is a pharmaceutical treatment used for an increasing number of neurological and non-neurological indications, symptoms and diseases. Despite the wealth of clinical reports that involve the timing of the therapeutic effects of this toxin, few studies have attempted to integrate these data into unified models. Secondary reactions have also been examined including the development of adverse events, resistance to repeated applications, and nerve terminal sprouting. Our primary intent for conducting this review was to gather relevant pharmacodynamic data from suitable biomedical literature regarding botulinum neurotoxins via the use of automated data-mining techniques. We envision that mathematical models will ultimately be of value to those who are healthcare decision makers and providers, as well as clinical and basic researchers. Furthermore, we hypothesize that the combination of this computer-intensive approach with mathematical modeling will predict the percentage of patients who will favorably or adversely respond to this treatment and thus will eventually assist in developing the increasingly important area of personalized medicine. PMID- 20021329 TI - Addressing the challenge: current and future directions in ovarian cancer therapy. AB - Numerous ovarian gene therapy strategies are in clinical phases based on concepts of replacement/ knock out of deregulated gene, suicide gene strategies, strengthening of the immune response against a tumor, inhibition of tumor angiogenesis and growth factors. Non-viral delivery systems have potential advantages over currently widely used viral vectors and other classical vectors for delivering therapeutic gene of interest. The present review provides a comprehensive overview of potential of various delivery systems currently in use. Non-viral formulations used in ovarian gene therapy include injecting naked DNA, liposomes, polyplexes, lipopolyplexes, nanoparticles, gene gun and ultrasound/microbubble mediated gene delivery. In addition to improving vector delivery, the DNA constructs need to be optimised for both efficient and long term transgene expression. Minicircles using minimal immunological defined gene expression (MIDGE) technology, are a promising future alternative to plasmid for use in non-viral ovarian gene therapy in terms of biosafety, improved gene transfer, potential bioavailability, minimal size and little immune reaction. The review explores the best route of administration for ovarian cancer gene therapy given its peritoneal dissemination which poses a major challenge in treating ovarian cancer patients. Enhancement of therapeutic index can be further achieved by overcoming barriers both at cellular and nuclear levels. Selective tumor targeting with minimal toxicity using folate modified, incorporating nuclear localization signal and PEGylated stealth liposome's represents a popular approach and needs to be exploited in ovarian gene therapy. PMID- 20021330 TI - State-of-the-art lentiviral vectors for research use: risk assessment and biosafety recommendations. AB - Lentiviral vectors (LV) are competent gene transfer vehicles, as used for both research and gene therapy applications, because of their stable integration in non-dividing and dividing cells and long-term transgene expression. Along with our understanding that LV offer solutions for gene therapy, biosafety concerns have uncovered risks due to insertional mutagenesis, the generation of replication competent lentiviruses (RCL) and vector mobilization. Researchers therefore continue to devote significant efforts in designing LV with improved efficacy and biosafety features. The choice of a particular LV system for experimental studies is often driven by functional considerations, including increased productivity and/or transduction efficiency. The design of safer vectors has also directly benefited researchers allowing them to conduct experimental studies with lower risk. Currently, vectors combine improved safety features (that decrease the risk of recombination and vector mobilization) with increased transduction efficiency. Hence, risks associated with the inadvertent transduction of cells of the investigator gain greater importance in assessing the overall risk of these vectors and become an important biosafety concern. This review outlines the different strategies used to improve LV biosafety by comparing state-of-the-art and emerging LV production systems and highlighting biosafety issues that can arise during their contained use. The few existing national and international biosafety recommendations that specifically address the use of LV in research are discussed and recommendations for most common research activities using LV are proposed. PMID- 20021331 TI - Targeted nucleic acid delivery to mitochondria. AB - Mitochondrial genetics has become an emerging area of research in the field of modern therapeutics. Mitochondrial genome is the source of 13 polypeptides which are components of subunits of complexes of electron transport chain and are used in the generation of ATP by oxidative phosphorylation. Any mutation and/or defects in these mitochondrial genes may cause diseases ranging from neurodegenerative diseases, diabetes mellitus to cancer. In an ideal condition mtDNA should be mutation free. There are various mechanisms for the repair of diseased cell or mitochondrial DNA. Nowadays, nucleic acid based therapeutics has become of interest and represent a new area of research. However, problem consistently encountered is safe and effective delivery of DNA to the mitochondria for therapeutic benefits. There are numerous barriers which are to be surpassed for successful delivery of nucleic acid to the cell interior and ultimately to the mitochondria. For efficient and effective DNA delivery to the mitochondrial matrix, a suitable carrier system is required to be designed and developed. In the present review we have discussed briefly about mitochondrial DNA and related diseases, various barriers encountered in the delivery of DNA, internalization processes, delivery strategies and methods for targeted delivery of DNA to the mitochondria. PMID- 20021332 TI - Central nervous system (CNS)-related pathologies. PMID- 20021333 TI - Creutzfeldt-jakob, Parkinson, lewy body dementia and Alzheimer diseases: from diagnosis to therapy. AB - Depositions of proteins in form of amyloid and non-amyloid plaques are common pathogenic signs of more than 20 degenerative diseases affecting the central nervous system or a variety of peripheral tissues. Among the neuropathological conditions, Alzheimer's, Parkinson's and the prion diseases, such as Creutzfeldt Jakob disease (CJD), present ambiguities as regarding their differential diagnosis. At present, their diagnosis must be confirmed by post-mortem examination of the brain. Currently the ante-mortem diagnosis is still based on the integration of multiple data (clinical, paraclinical and biological analyses) because no unique marker exists for such diseases. The detection of specific biomarkers would be useful to develop a differential diagnostic, distinguishing not only different neurodegenerative diseases but also the disease from the non pathological effects of aging. Several neurodegenerative biomarkers are present at very low levels during the early stages of the disease development and their ultra-low detection is needed for early diagnosis, which should permit more effective therapeutic interventions, before the disease concerned can progress to a stage where considerable damage to the brain has already occurred. In the case of prion diseases, there are concerns regarding not only patient care, but the wider community too, with regard to the risk of transmission of prions, especially during blood transfusion, for which, four cases of variant CJD infection associated with transfusion of non-leukocyte-depleted blood components have been confirmed. Therefore the development of techniques with high sensitivity and specificity represent the major challenge in the field of the protein misfolding diseases. In this paper we review the current analytical and/or biochemical diagnostic technologies used mainly in prion, but also in Alzheimer and Parkinson diseases and emphasizing work on the protein detection as a surrogates and specific biomarker in the body fluid of patients (urine, CSF and blood). This review highlights the urgency of the development of early and sensitive diagnostics in terms of therapeutic challenge. PMID- 20021334 TI - Anti-inflammatory effects of antidepressants: possibilities for preventives against Alzheimer's disease. AB - Increasing evidence of pro-inflammatory mediator expression in major depressions indicate that inflammatory changes may play a role. If this is true, the efficacy of antidepressants may be partially attributable to suppression of inflammation. Various types of antidepressants can suppress serum and plasma levels of pro inflammatory mediators in patients with major depression. Therefore they can inhibit the production of pro-inflammatory mediators by immune cells. These include glial cells, which are the main sources and targets of cytokines in the brain. This review summarizes the evidence showing that antidepressants have an anti-inflammatory potential. The putative mechanisms are also discussed. Because of the anti-inflammatory effects of antidepressants, they might also act as preventives for neurodegenerative dementias including Alzheimer's disease, where the pathogenesis involves chronic inflammation associated with activated microglia. PMID- 20021335 TI - Advances in the immune pathogenesis and treatment of multiple sclerosis. AB - Multiple sclerosis (MS) is a disorder of the central nervous system (CNS). It is characterized by episodic and progressive neurological dysfunction resulting from inflammatory and autoimmune reactions, myelin loss, conduction block, oligodendrocyte pathology, gliosis, and axonal loss in CNS. Recent years have witnessed advances in better understanding the immune pathogenesis of MS, prompted by animal models, human pathological observations and MRI studies. There have been significant changes in the therapeutic regimens in MS, with an emphasis on preventative treatment of an ongoing disease process. Agents in use and in the research pipeline have mechanisms that act on various anti-inflammatory and immunomodulatory properties, including blocking leukocyte migration into CNS and targeting chemoattraction. In addition, recent studies on the neurodegenerative components of MS have directed therapeutic trials to neuroprotection and neurorestoration. In this paper, we summarize the current understanding of the mechanisms of approved pharmacological agents and review the putative mechanisms and status of some important agents in clinical phase two or three trials in MS. PMID- 20021336 TI - Recent advances in the treatment of amyotrophic lateral sclerosis. Emphasis on kynurenine pathway inhibitors. AB - Amyotrophic lateral sclerosis (ALS) is an adult onset, progressive and fatal motor neuron degenerative disease [1]. The aetiology of ALS is currently unknown, though strongly suggested to be multifactorial. Recently, the kynurenine pathway (KP) has emerged as a potential contributing factor [2]. The KP is a major route for the metabolism of tryptophan, generating neuroactive intermediates in the process. These catabolites include the excitotoxic N-methyl-D-aspartate (NMDA) receptor agonist, quinolinic acid (QUIN) [3] and the neuroprotective NMDA receptor antagonist, kynurenic acid (KYNA) [4,5]. These catabolites appear to play a key role in the communication between the nervous and immune systems, and also in modulating cell proliferation and tissue function [6]. As the cause of ALS is still unknown, there is presently no efficient treatment for it. Currently, Riluzole is the drug of choice but its effect is relatively modest [7]. Targeting the KP, hence, could offer a new therapeutic option to improve ALS treatment [8]. Several drugs that block the KP are already under investigation by our laboratory and others, some of which are in or about to enter clinical trials for other diseases. For example, the KP inhibitors, Teriflunomide (Sanofi Aventis) and Laquinimod (Teva Neuroscience). Recently, a KP inhibitor has also reached the Japan market as an immunomodulative drug [9]: Tranilast/Rizaben (Angiogen Ltd.) is an anthranilic acid derivative [8]. Finally, the 8 hydroxyquinolinine metal attenuating compounds, Clioquinol and PBT2, interestingly have close structural similarity with KYNA and QUIN. Such drugs would open a new and important therapeutic door for ALS. PMID- 20021337 TI - Neurotrophic factor treatment after spinal root avulsion injury. AB - Spinal root avulsion injury causes motoneuron death and immediate loss of sensory and motor functions. Surgical intervention such as reimplantation of avulsed root is proven useful to restore neural circuitry of spinal cord and targeted muscles. Yet, additional strategies are required for faster and better functional recovery which is overall unsatisfactory. Accumulating evidences in animal studies, particularly in peripheral nerve injuries, demonstrated the effectiveness of neurotrophic factors in rescuing injured motoneurons and promoting axon regeneration. It is, however, important to recognize the differences between peripheral nerve and avulsion injury. In this review, we will briefly describe the changes in motoneurons after avulsion and provides a comprehensive list of neurotrophic factors which are known to exert neuroprotective effects on motoneurons. We will include recent studies on trophic factors for motoneuron survival and regeneration in peripheral nerve and avulsion injuries. We will also discuss the potential use of trophic factors in the context of avulsion injuries. PMID- 20021338 TI - Treatment of viral encephalitis. AB - Several viruses may cause central nervous system diseases with a broad range of clinical manifestations. The time course of the viral encephalitis can be acute, subacute, or chronic. Pathologically there are encephalitis with direct viral entry into the CNS in which brain parenchyma exhibits neuronal damaging and viral antigens and there are postinfectious autoimmune encephalitis associated with systemic viral infections with brain tissue presenting perivascular aggregation of immune cells and myelin damaging. Some virus affect previously healthy individuals while others produce encephalitis among imunocompromised ones. Factors such evolving lifestyles and ecological changes have had a considerable impact on the epidemiology of some viral encephalitis [e.g. West-Nile virus, and Japanese B virus]. Citomegalovirus and JC virus are examples of infections of the brain that have been seen more frequently because they occur in immunocompromised patients. In the other hand many scientific achievements in neuroimaging, molecular diagnosis, antiviral therapy, immunomodulatory treatments, and neurointensive care have allowed more precise and earlier diagnoses and more efficient treatments, resulting in improved outcomes. In this article, we will present the current drug options in the management of the main acute and chronic viral infection of the central nervous system of immunocompetent and immunocompromised adults, focusing on drugs mechanisms of action, efficacy, and side effects. The early diagnosis and correct management of such diseases can reduce mortality and neurological sequelae; however, even with recent treatment advances, potentially devastating outcomes are still possible. PMID- 20021339 TI - Neuroleptics and migraine. AB - Many dopamine antagonists are proven acute migraine treatments. Genetic studies also imply that polymorphisms in dopamine genes (DRD2 receptors) in persons with migraine may create dopamine hypersensitivity. However, treatment is limited by the adverse event profiles of conventional neuroleptics including extrapyramidal symptoms, anticholinergic and antihistaminergic effects, hyperprolactinemia, and prolonged cardiac QT interval. Atypical neuroleptics cause less extrapyramial symptoms and some atypical neuroleptics, including olanzapine and quetiapine, may be beneficial as both acute and preventive migraine treatment. The combination of prochlorperazine, indomethacin, and caffeine is effective in the treatment of the acute migraine attack. The mechanism of action by which neuroleptics relieve headache is probably related to dopamine D2 receptor antagonist. Other actions via serotonin (5HT) receptor antagonists may also be important, particularly for migraine prevention. Additional studies to clarify the mechanism of action of neuroleptics in migraine could lead to new drugs and better management of migraine. PMID- 20021340 TI - Mechanisms and treatment of neuropathic pain. AB - Neuropathic pain (pain associated with lesions or dysfunction of nervous system) is relatively common, occurring in about 1% of the population. Studies in animal models describe a number of peripheral and central pathophysiological processes after nerve injury that would be the basis of underlying neuropathic pain mechanism. A change in function, chemistry, and structures of neurons (neural plasticity) underlie the production of the altered sensitivity characteristics of neuropathic pain. Peripheral sensitization acts on the nociceptors, and central sensitization takes place at various levels ranging from the dorsal horn to the brain. In addition, abnormal interactions between the sympathetic and sensory pathways contribute to mechanisms mediating neuropathic pain. Despite recent advances in identification of peripheral and central sensitization mechanisms related to nervous system injury, the effective treatment of patients suffering from neuropathic pain remains a clinical challenge. Although numerous treatment options are available for relieving neuropathic pain, there is no consensus on the most appropriate treatment. However, recommendations can be proposed for first-line, second-line, and third-line pharmacological treatments based on the level of evidence for the different treatment strategies. Beside opioids, the available therapies shown to be effective in managing neuropathic pain include anticonvulsants, antidepressants, topical treatments (lidocaine patch, capsaicin), and ketamine. Tricyclic antidepressants are often the first drugs selected to alleviate neuropathic pain (first-line pharmacological treatment). Although they are very effective in reducing pain in several neuropathic pain disorders, treatment may be compromised (and outweighed) by their side effects. In patients with a history of cardiovascular disorders, glaucoma, and urine retention, pregabalin and gabapentine are emerging as first-line treatment for neuropathic pain. In addition these anti-epileptic drugs have a favourable safety profile with minimal concerns regarding drug interactions and showing no interference with hepatic enzymes. Despite the numerous treatment options available for relieving neuropathic pain, the most appropriate treatment strategy is only able to reduce pain in 70% of these patients. In the remaining patients, combination therapies using two or more analgesics with different mechanisms of action may also offer adequate pain relief. Although combination treatment is clinical practice and may result in greater pain relief, trials regarding different combinations of analgesics are lacking (which combination to use, occurrence of additive or supra-additive effects, sequential or concurrent treatment, adverse-event profiles of these analgesics, alone and in combination) are lacking. Additionally, 10% of patients still experience intractable pain and are refractory to all forms of pharmacotherapy. If medical treatments fail, invasive therapies such as intrathecal drug administration and neurosurgical interventions may be considered. PMID- 20021341 TI - New antiepileptic drugs: molecular targets. AB - In the past 20 years, a number of new antiepileptic drugs (AEDs) have been introduced and other molecules are in development, some of which are advantageous in terms of pharmacokinetics, tolerability and potential for drug interactions. These drugs are regarded as second generation compared to older agents such as barbiturates, phenytoin, carbamazepine, ethosuximide and valproic acid. Although some of these second generation compounds may be advantageous in terms of kinetics, tolerability and potential for drug interactions, all of them still target voltage-gated channels or GABA-mediated inhibition, predominantly, without any real improvement in epilepsy therapy. Studies on mechanisms of seizure generation and propagation have identified new potential targets for AEDs. The growing understanding of the pathophysiology of epilepsy and the structural and functional characterization of the molecular targets provide many opportunities to create improved epilepsy therapies. In this review the molecular targets for new AEDs are discussed, providing further suggestions on how future research can be improved. PMID- 20021342 TI - Estrogens as potential therapeutic agents in multiple sclerosis. AB - The disease activity of multiple sclerosis (MS) is known to be ameliorated during pregnancy, and pregnancy is also found to be protective in experimental autoimmune encephalomyelitis (EAE), an animal model of MS. Estrogen levels increase during pregnancy and basic researches have shown that estrogens have immunomodulatory effects on immune cells. The importance of estrogen in pathogenic autoimmune diseases has also been demonstrated in EAE by altering hormone levels. Mice treated with estrogen experienced significantly decreased EAE severity and delayed onset of disease as a result of neuroprotective and anti inflammatory effects. Brain atrophy has been detected at the early stages of MS by using MRI; thus, as a neuroprotective agent, estrogen might be effective against brain atrophy. Estrogen's effects are primarily mediated by the nuclear estrogen receptor (ER), and recent studies have shown the presence of nuclear ERs on the cells involved in the immune response. There have been some reports on genetic polymorphisms of ERs in MS. In this review paper, we discuss increasing evidence that points to a link between estrogen and MS. We also analyze the therapeutic potential of estrogens in MS and review current genetic studies on ER. PMID- 20021343 TI - The insulin-like growth factor-1 axis and its potential as a therapeutic target in central nervous system (CNS) disorders. AB - The insulin-like growth factor-1 (IGF-1) is a pleiotropic factor. Many studies have revealed its importance in the development and maintenance of the central nervous system (CNS). This review will discuss the IGF-1 axis, from the factor itself to the signalling pathways it activates, and its tight regulation. Particular focus will be brought on potential therapeutic targets of the IGF-1 axis in CNS disorders, including brain tumours and neurodegenerative diseases affecting neurons and oligodendrocytes. PMID- 20021344 TI - Endogenous regulation of neural stem cells in the adult mammalian brain. AB - Tissue-specific stem cells replenish organs by replacing cells lost due to tears and wears or injury throughout life. Long considered as an exception to this rule, the adult mammalian brain has consistently been found to possess stem cells that ensure neurogenesis. Neural stem cells persist within the subventricular zone bordering the lateral ventricles of the brain. Constitutively, neural stem cells proliferate and produce a continuous supply of new neurons that migrate towards the olfactory bulb where they ensure turnover of interneurons. Owing to their potential clinical use for the treatment of neurodegenerative diseases, the factors that control proliferation, self-renewal and differentiation of neural stem cells have received increasing interest. These studies have unraveled that the cellular dynamic within the subventricular zone is tightly controlled by astrocytes and endothelial cells that neighbor neural stem cells. These neighboring cells produce substrate-bound and soluble factors that make up a specialized microenvironment named the neurogenic niche. The equilibrium between neural stem cells activity and quiescence is adjusted by neurons located in remote brain areas that adapt neuron production to physiological and pathological constraints. Brain injury or neurodegenerative diseases affect neural stem cells proliferation, differentiation and migration suggesting that neural stem cells are involved in brain self-repair. Understanding the endogenous mechanisms that regulate neural stem cells will help to replenish cellular constituents lost by injury and thereby allow an effective development of neural stem cells based therapies of brain diseases. PMID- 20021345 TI - Biochemistry and neurobiology of prosaposin: a potential therapeutic neuro effector. AB - Prosaposin, a 66 kDa glycoprotein, was identified initially as the precursor of the sphingolipid activator proteins, saposins A-D, which are required for the enzymatic hydrolysis of certain sphingolipids by lysosomal hydrolases. While mature saposins are distributed to lysosomes, prosaposin exists in secretory body fluids and plasma membranes. In addition to its role as the precursor, prosaposin shows a variety of neurotrophic and myelinotrophic activities through a receptor mediated mechanism. In studies in vivo, prosaposin was demonstrated to exert a variety of neuro-efficacies capable of preventing neuro-degeneration following neuro-injury and promoting the amelioration of allodynia and hyperalgesia in pain models. Collective findings indicate that prosaposin is not a simple house keeping precursor protein; instead, it is a protein essentially required for the development and maintenance of the central and peripheral nervous systems. Accumulating evidence over the last decade has attracted interests in exploring and developing new therapeutic approaches using prosaposin for human disorders associated with neuro-degeneration. In this review we detail the structure characteristics, cell biological feature, in vivo efficacy, and neuro-therapeutic potential of prosaposin, thereby providing future prospective in clinical application of this multifunctional protein. PMID- 20021346 TI - Histamine H3-receptor inverse agonists as novel antipsychotics. AB - Schizophrenia (SZ) that is resistant to treatment with dopamine (DA) D2 antagonists may involve changes other than those in the dopaminergic system. Recently, histamine (HA), which regulates arousal and cognitive functions, has been suggested to act as a neurotransmitter in the central nervous system. Four HA receptors-H1, H2, H3, and H4-have been identified. Our recent basic and clinical studies revealed that brain HA improved the symptoms of SZ. The H3 receptor is primarily localized in the central nervous system, and it acts not only as a presynaptic autoreceptor that modulates the HA release but also as a presynaptic heteroreceptor that regulates the release of other neurotransmitters such as monoamines and amino acids. H3-receptor inverse agonists have been considered to improve cognitive functions. Many atypical antipsychotics are H3 receptor antagonists. Imidazole-containing H3-receptor inverse agonists inhibit not only cytochrome P450 but also hERG potassium channels (encoded by the human ether-a-go-go-related gene). Several imidazole H3-receptor inverse agonists also have high affinity for H4 receptors, which are expressed at high levels in mast cells and leukocytes. Clozapine is an H4-receptor agonist; this agonist activity may be related to the serious side effect of agranulocytosis caused by clozapine. Therefore, selective non-imidazole H3-receptor inverse agonists can be considered as novel antipsychotics that may improve refractory SZ. PMID- 20021347 TI - Therapeutic strategies in HTLV-I-associated myelopathy/tropical spastic paraparesis (HAM/TSP). AB - Human T lymphotropic virus type I (HTLV-I)-associated myelopathy/tropical spastic paraparesis (HAM/TSP) is chronic progressive myelopathy characterized by bilateral pyramidal tracts involvement with sphincteric disturbances. HTLV-I infects approximately 10-20 million people worldwide. There are large endemic areas in southern Japan, the Caribbean, Central and South America, the Middle East, Melanesia, and equatorial regions of Africa. Since the primary neuropathological feature of HAM/TSP is chronic inflammation caused by HTLV-I infection in the spinal cord, various treatments focusing on immunomodulatory or anti-viral effects were performed for HAM/TSP patients until now. However, there are still many of problems, such as insufficient effects, side effects and expensive costs in long-term treatments, etc., in these treatments. Therefore, an ideal therapeutic strategy against HAM/TSP is still not established yet. Although only a small proportion of HTLV-I-infected individuals develops HAM/TSP, neurological symptoms are certainly progressive once myelopathy develops, leading to deterioration of the quality of life. Therefore, we now need the therapeutic regimens to protect the development, or be able to commence the treatments as soon as possible after the development safely and inexpensively even in long-term course or lifelong course of treatment. As HTLV-I-infected CD4(+) T cells are the first responders in the immunopathogenesis of HAM/TSP, the ideal treatment is the elimination of HTLV-I-infected cells from the peripheral blood. In this article, we will review the therapeutic strategies against HAM/TSP up to now and will introduce our new therapeutic approach focusing on the targeting of HTLV-I infected cells in HAM/TSP patients. PMID- 20021348 TI - The trigeminal system in birds and nociception. AB - Aim of this paper is to give a concise overview of what is known about the trigeminal nociceptive system in birds. Several types of nociceptors have been discovered, thermal nociceptors and polymodal, i.e. mechanothermal and mechanochemical receptors. Information from these receptors reaches the Laminae I and II of the caudal subnucleus of the descending trigeminal system and of the dorsal horn of the rostral spinal cord. The organization of the afferents to the avian brainstem and of the primary nociceptive centers is largely the same as that in mammals. This is also true for a number of histochemical characteristics of these primary centers. The comparability of the ascending nociceptive system in birds and mammals is more problematic. This is due to the differences in organization of the forebrain in mammals and birds. The paper concludes with a short discussion on the sense of pain and the connection with nociception. PMID- 20021349 TI - The enigmatic sigma receptors. PMID- 20021350 TI - In vitro and ex vivo characterization of sigma-1 and sigma-2 receptors: agonists and antagonists in biological assays. AB - Methods for addressing sigma receptor affinity and activity have been explored and although several protocols have been employed, only few procedures resulted reliable. Sigma-1 receptor affinity protocol using guinea-pig brain and (+) [(3)H]-pentazocine and sigma-2 receptor affinity protocol employing rat liver and [(3)H]-DTG are usually reported by authors as standard procedures. By contrast, the intrinsic activity evaluation of sigma ligands has been performed in several manners: tumor cell lines, isolated organ bath, in vivo animal model. The last is not considered in the present paper because this method studied the physiological role of sigma receptors. The studies carried out in tumor cell lines involved the role of sigma receptors in tumors progression while, although isolated organ bath experiment employed physiological samples, the pharmacokinetic properties of ligands, a strictly requirement for the in vivo assays, did not affect the pharmacodynamic properties of tested compounds. The advances in the above mentioned assays have been reported. PMID- 20021351 TI - Selective sigma-1 (sigma1) receptor antagonists: emerging target for the treatment of neuropathic pain. AB - A large number of therapeutic roles have been proposed for sigma(1) receptors but the involvement of sigma(1) receptor in non-acute pain had not been well explored up to now. sigma(1) receptor knock-out mice became available offering us the possibility to study the role of sigma(1) receptor in nociception, particularly in models where central sensitization processes play a significant role. Given the attractive therapeutic potential, we have developed a chemical program aimed at the discovery of novel and selective sigma(1) ligands. Herein we discuss the rational basis of this approach and report preliminary pharmacological results of several chemical series and aspects of their structure-activity relationship on sigma(1) receptor. Functional data in pain models are presented mainly on one series that provide evidence to consider selective sigma(1) receptor antagonists an innovative and alternative approach for treating neuropathic pain. PMID- 20021353 TI - Imaging of sigma1 receptors in the human brain using PET and [11C]SA4503. AB - Sigma(1) receptors were imaged in living human brain by positron emission tomography (PET) using [(11)C] SA4503. A dynamic 90-min scan and kinetic analysis enabled quantification of receptor density in the brain. The sigma(1) receptors were distributed throughout the brain in normal subjects, but decreased in the frontal, temporal, and occipital lobes, cerebellum and thalamus in patients with early Alzheimer's disease and in the putamen in patients with Parkinson's disease. In addition, rates of receptor occupancy by the neuroleptic haloperidol and the selective serotonin reuptake inhibitor fluvoxamine were evaluated by [(11)C]SA4503-PET and found to be high. [(11)C]SA4503-PET is useful for studying the pathophysiology of neurological and psychiatric disorders such as schizophrenia and for evaluation of the pharmacodynamics of psychiatric drugs. PMID- 20021352 TI - Sigma-1 receptor chaperones and diseases. AB - Chaperones are proteins that assist the correct folding of other protein clients either when the clients are being synthesized or at their functional localities. Chaperones are responsible for certain diseases. The sigma-1 receptor is recently identified as a receptor chaperone whose activity can be activated/deactivated by specific ligands. Under physiological conditions, the sigma-1 receptor chaperones the functional IP3 receptor at the endoplasmic reticulum and mitochondrion interface to ensure proper Ca(2+) signaling from endoplasmic reticulum into mitochondrion. However, under pathological conditions whereby cells encounter enormous stress that results in the endoplasmic reticulum losing its global Ca(2+) homeostasis, the sigma-1 receptor translocates and counteracts the arising apoptosis. Thus, the sigma-1 receptor is a receptor chaperone essential for the metabotropic receptor signaling and for the survival against cellular stress. The sigma-1 receptor has been implicated in many diseases including addiction, pain, depression, stroke, and cancer. Whether the chaperone activity of the sigma-1 receptor attributes to those diseases awaits further investigation. PMID- 20021354 TI - Sigma-1 receptors and selective serotonin reuptake inhibitors: clinical implications of their relationship. AB - Endoplasmic protein sigma-1 receptors represent unique binding sites in the brain, and they exert a potent influence on a number of neurotransmitter systems. Several lines of evidence suggest that sigma-1 receptors play roles in the pathophysiology of psychiatric diseases, as well as in the active mechanisms of some selective serotonin reuptake inhibitors (SSRIs). Interestingly, we reported that some SSRIs possess moderate to high affinities at sigma-1 receptors in the brain. Among them, the order of affinity for sigma-1 receptors was as follows: fluvoxamine > sertraline > fluoxetine > citalopram " paroxetine. In a cell culture system, we demonstrated that fluvoxamine, but not sertraline or paroxetine, significantly potentiated nerve-growth factor (NGF)-induced neurite outgrowth in PC12 cells, and that the effect of fluvoxamine on NGF-induced neurite outgrowth was significantly antagonized by treatment with the selective sigma-1 receptor antagonist NE-100. Furthermore, we reported that phencyclidine (PCP)-induced cognitive deficits in mice were significantly improved by subsequent subchronic administration of fluvoxamine, but not sertraline and paroxetine, and that the effect of fluvoxamine on PCP-induced cognitive deficits was antagonized by co-administration of NE-100. Moreover, a recent study using the specific sigma-1 receptor ligand [(11)C] SA4503 and positron emission tomography (PET) have demonstrated that an oral administration of fluvoxamine, but not paroxetine, could bind to sigma-1 receptors in the healthy human brain, in a dose-dependent manner. These findings suggest that sigma-1 receptors might be implicated in the active mechanisms of fluvoxamine. In this article, the author would like to discuss the novel role of sigma-1 receptors in the active mechanisms of some SSRIs including fluvoxamine. PMID- 20021355 TI - 1-Cyclohexylpiperazine and 3,3-dimethylpiperidine derivatives as sigma-1 (sigma1) and sigma-2 (sigma2) receptor ligands: a review. AB - Herein the evolution in the development of new sigma (sigma) receptor ligands since the middle '90s by our research group is reported. In the effort to contribute to the identification of the structural features for high-affinity ligands selective versus serotonin, dopamine and other CNS-related receptors, two general classes of (naphthalene)alkylamine compounds were prepared and explored, with the aim of addressing the affinities toward the two recognized sigma receptor subtypes. The common template of these compounds was mainly an unsubstituted or methoxy-substituted naphthalene or tetralin nucleus, linked by an alkyl spacer to a substituted piperazine or piperidine ring. The design of new ligands was thought keeping in mind their possible application as PET diagnostic tools and fluorescence tools. High-affinity sigma(2) receptor ligands were found among N-cyclohexylpiperazine derivatives, such as 1-cyclohexyl-4-[3-(5-methoxy 1,2,3,4-tetrahydronaphthalen-1-yl)propyl]piperazine (3) (PB 28), when they were assayed in radioligand binding with [(3)H]-DTG in rat liver. Unfortunately, these ligands were all devoid of a significant selectivity relative to sigma(1) receptor whose binding was assayed with (+)-[(3)H]-pentazocine in guinea pig brain. Nevertheless, compound 3 had previously shown to be 40-fold selective with a slightly different binding method in animals' tissues. Moreover, it demonstrated 46-fold and 59-fold sigma(2) versus sigma(1) receptor binding selectivity in MCF7 and MCF7 ADR tumor cell lines respectively. In the class of piperazines, also high-affinity sigma(1) receptor ligands were found, possibly due to the presence of a double N-atom and an additional reverse mode of binding. Piperidine derivatives were investigated as high-affinity and selective sigma(1) receptor ligands leading to some 3,3-dimethylpiperidines such as 3,3-dimethyl-1 [3-(6-methoxynaphthalen-1-yl)propyl]piperidine (69) which resulted to be highly selective relative to the sigma(2) receptor. For the best ligands, functional assays were conducted in order to investigate agonist/antagonist activity. The effect of chirality in the intermediate methyl-alkyl chain was explored for a class of 4-methylpiperidines linked to some (4-chlorophenoxy)alkyl moieties, and compound (-)-(S)-92 emerged as the most selective sigma(1) relative to sigma(2) receptor ligand. PMID- 20021356 TI - Structure-affinity-relationship study of bicyclic sigma receptor ligands. AB - It was postulated that N(6)-allyl bicyclic derivatives 1 bind with N-8 at the proton donor site of the sigma(1) receptor and that a substituent in 2-position of the bicyclic framework 1 results in unfavorable steric interactions with the sigma(1) receptor protein. In order to support this hypothesis both enantiomers of 6-allyl-8-(4-methoxybenzyl)-6,8-diazabi-cyclo[3.2.2]non-2-ene (2/ent-2) and 6 benzyl-8-(4-methoxybenzyl)-6,8-diazabicyclo[3.2.2]nonane 3/ent-3 were synthesized stereoselectively. The (S,S)-configured enantiomers 2 and 3 are the eutomers with eudismic ratios of 31 and 4.8, respectively. Therefore, these enantiomers are used in the sigma(1) pharmacophore model. The N(6)-allyl derivative 2 with a double bond in the three carbon bridge adopts the orientation 2c with N-8 interacting with the sigma(1) receptor proton donor site (Fig. 2) resulting in slightly reduced steric interactions of the small double bond in 2/3-position. The almost C(2)-symmetric benzyl derivative 3 can adopt both orientations 2c and 2d at the sigma (1) receptor (N-8 or N-6 interacts with the sigma (1) receptor proton donor site) resulting in subnanomolar sigma(1) receptor affinity (K(i) = 0.91 nM). PMID- 20021358 TI - A structure-affinity and comparative molecular field analysis of sigma-2 (sigma2) receptor ligands. AB - Several sigma(1) receptor ligands with sub-nanomolar affinity and excellent selectivity have been reported, but relatively few sigma(2)-selective ligands are known. 1-Cyclohexyl-4-[3-(5-methoxy-1,2,3,4-tetrahydronaphthalen-1-yl)propyl] piperazine (PB28; 1) has been reported by us as a high-affinity sigma(2) receptor ligand with significant sigma(2) selectivity, and several analogs of (1) now have been developed. Among these are the class of cyclohexylpiperazines that display a good compromise between affinity/activity and selectivity for sigma(2) receptors. Very little is currently known about the nature of sigma(2) receptors. In the absence of structure-based receptor information, we applied a comparative molecular field analysis (CoMFA) - a three-dimensional structure-activity relationship (3D-QSAR) method - to a set of cyclohexylpiperazine sigma(2) ligands to develop a predictive model that might provide information about the stereoelectronic nature of the receptor binding site. Two CoMFA models were generated from two different alignments: the first used an automated FlexS algorithm, and the second used a rationally-driven manual alignment. Significantly better predictivity was obtained with the manual alignment (TSET: q(2) = 0.73, r(2) = 0.95; PSET: r(2) = 0.55/0.73) than from the automated alignment (TSET: q(2) = 0.69, r(2) = 0.98; PSET: r(2) = 0.13/0.16). The resulting CoMFA maps account for observed structure-affinity relationships and suggest a possible anatomy for the sigma(2) receptor/cyclohexylpiperazine binding site. PMID- 20021357 TI - Development of molecular probes for imaging sigma-2 receptors in vitro and in vivo. AB - The sigma-2 (sigma(2)) receptor is proving to be an important protein in the field of cancer biology. The observations that sigma(2) receptors have a 10-fold higher density in proliferating tumor cells than in quiescent tumor cells, and that sigma(2) receptor agonists are capable of killing tumor cells via apoptotic and non-apoptotic mechanisms, indicate that this receptor is an important molecular target for the development of radiotracers for imaging tumors using techniques such as Positron Emission Tomography (PET) and Single Photon Emission Computed Tomography (SPECT) and for the development of cancer chemotherapeutic agents. In spite of recent promising results towards achieving these goals, research in this field has been hampered by the fact that the molecular identity of the protein sequence of the sigma(2) receptor is currently not known. Consequently, most of what is known about this protein has been obtained using either radiolabeled or fluorescent probes for this receptor, or biochemical analysis of the effect of sigma(2) selective ligands on cells growing under tissue culture conditions. This article provides a review of the development and use of sigma(2) receptor ligands, and how these ligands have been used with a variety of in vitro and in vivo models to gain a greater understanding of the role this receptor plays in cancer. PMID- 20021359 TI - Brain serotonin, psychoactive drugs, and effects on reproduction. AB - Serotonin, a biogenic amine, is present in significant amounts in many structures of the CNS. It is involved in regulation of a wide variety of physiological functions, such as sensory and motor functions, memory, mood, and secretion of hormones including reproductive hormones. It has also been implicated in the etiology of a range of psychiatric disorders such as anxiety, depression, and eating disorders, along with other conditions such as obesity and migraine. While some drugs that affect serotonin, such as fenfluramine and fluoxetine, have been successfully used in treatment of a range of psychiatric diseases, others, such as the amphetamine analogues MDMA and METH, are potent psychostimulant drugs of abuse. Alterations in serotonergic neurons caused by many of these drugs are well characterized; however, little is known about the reproductive consequences of such alterations. This review evaluates the effects of drugs such as MDMA, pCA, fenfluramine, and fluoxetine on serotonergic transmission in the brain, examines the relationships of these drug effects with the neuroendocrine mechanisms modulating reproductive events such as gonadotropin secretion, ovulation, spermatogenesis, and sexual behavior in animal models, and discusses possible reproductive implications of these drugs in humans. PMID- 20021360 TI - On some physiological aspects of ethanol repercussion on neural and cardiorenal functions. AB - Chronic ethanol ingestion, mostly in young adults, constitutes a frequent drug abuse situation, which is associated to a wide variety of pathological disturbance affecting a number of organs, including liver, kidney, heart, pancreas and brain. The ethanol effects are more prominent when occurring at the perinatal period of life, generating, among other disabilities, brain developmental and functional impairments, as well as the so-called "fetal alcoholic syndrome". However, low doses of ethanol, although not producing conspicuous signs of physiological impairment, may affect the developing organism, impairing the renal and cardiovascular system, among others. As a consequence of increased oxidative stress produced by ethanol intake and its subsequent oxidation, lipid peroxidation increases, enhancing reactive oxygen species formation, which is potentially injurious to the brain tissue. When occurring during gestation, lipid peroxidation may occur in the placenta, an event that would partially be responsible for fetal nutrition disturbance and consequently late physiological impairment. In this short review, data on ethanol effects on the nervous and cardiorenal structure and function are analyzed at the light of the most relevant hypotheses concerning ethanol mechanisms of action. Additionally, experimental data from the authors' laboratories are presented and discussed, focusing particular attention to the possibility of differential neural and cardiorenal ethanol effects as a function of the dose used in distinct experimental models. PMID- 20021361 TI - Dual cross-talk between nitric oxide and D-serine in astrocytes and neurons in the brain. AB - The present review describes the role of the putative cross-talk between two neurotransmitters, nitric oxide (NO) and D-serine, in the brain. Under physiological conditions NO homeostasis guarantees the correct function of NO in a number of events in the brain such as neurotransmission and vascular tone regulation. D-serine, produced in astrocytes, acts synergistically with glutamate at NMDA receptors on postsynaptic neurons. Neuronal and endothelial NO synthase (nNOS and eNOS) in astrocytes cross-talk with serine racemase (SR) and D-amino acid oxydase (DAAO), catalyzing the synthesis and degradation of D-serine, respectively. SR is inhibited by NO which activates DAAO. D-serine inhibits nNOS but not eNOS and activates SR. Astrocytes and neurons also cross-talk through NO/D-serine system. D-serine released from astrocytes induces a rapid increase in NO contents in postsynaptic neurons. Overall, D-serine production in astrocytes is negatively regulated by NO. Under inflammatory conditions, pro-inflammatory cytokines or Abeta induce, first, a drop in NO contents and an increase in the amounts of D-serine in astrocytes. Together with enhanced glutamate release from presynaptic neurons, D-serine induces an increase in Ca(2+) up-take into presynaptic neurons. In astrocytes an initial drop in NO contents triggers NF kappaB activation followed by inducible NOS (iNOS) expression. iNOS-derived massive amounts of NO may potentially be toxic. Under schizophrenic conditions, D serine production is down-regulated. Together with reduced glutamate release, this situation leads to the decreased NO production in postsynaptic neurons. In astrocytes induction of iNOS expression becomes predominant. Initial drop in nNOS derived NO is potentially toxic in this scenario. PMID- 20021362 TI - Anticonvulsant and neurotoxicity evaluation of some N(4) phenyl substituted pyridyl semicarbazones. AB - A series of 4-aryl substituted semicarbazones of pyridyl carbaldehyde and pyridyl methyl ketone were designed and synthesized to meet the structural requirements essential for anticonvulsant activity. The structure of synthesized compounds were confirmed by IR spectroscopy, PMR spectroscopy and nitrogen estimation. All the compounds were evaluated for anticonvulsant activity and neurotoxicity. Anticonvulsant activity was determined after intraperitoneal (i.p.) administration to mice by maximal electroshock (MES) and subcutaneous metrazol (ScMet) induced seizure methods and minimal motor impairment was determined by rotorod test. Majority of compounds exhibited significant anticonvulsant activity after intraperitoneal administration. The results obtained showed that 85.7 % of the compounds afforded protection in the MES test and 64.2 % of the total compounds afforded protection in ScPTZ test. Some of them also showed good activity after oral administration. In this study (Methyl-4- pyridyl) ketone N(4)- (p- chloro phenyl) substituted semicarbazone emerged as most active derivative showing activity at 100 mg/kg in both the test with prolonged duration of action. In the present study semicarbazones of pyridyl containing carbonyl compounds emerges as the lead molecule, showing broad spectrum of activity with low neurotoxicity and prolong duration of action on oral administration. Thus these may be utilized for the future development of novel anticonvulsants with broad spectrum of anticonvulsant activity. PMID- 20021363 TI - Efficacy of HGF gene transfer for various nervous injuries and disorders. AB - Hepatocyte growth factor (HGF) was originally identified as a molecule that could stimulate DNA synthesis in rat and human hepatocytes by autophosphorylation of the proto-oncogene c-met, which is a high-affinity receptor for HGF. Although it was at first considered that HGF could exert biological effects only on specific target cells, it has since been demonstrated that HGF mediates inflammatory responses to tissue injury and also regulates cell growth, cell motility, and morphogenesis in a wide variety of cell types, including cells within the nervous system. In the nervous system, HGF plays a role as a potent neurotrophic and angiogenetic factor. This factor promotes both the survival of neurons and the regeneration of injured nerves, and may also function as target-derived axonal chemoattractants, guiding axons to their target. These observations raised hopes that HGF protein might be useful for the clinical treatment of nervous system disorders. However, administration of HGF as a recombinant protein is still beset by a number of problems, such as a short serum half-life and poor access to the central nervous system by the systemic route because of the presence of the blood brain barrier. These problems can be major obstacles to the clinical use of this factor in a recombinant protein form, and has highlighted the need to develop innovative therapeutic strategies for more efficient delivery into the nervous system. Gene transfer into the nervous system has enormous therapeutic potential for a wide variety of disorders. It appears to have advantages over the administration of single or multiple bolus doses of a recombinant protein because gene transfer can achieve an optimally high, local concentration within the nervous system. In this article, we demonstrate the efficacy of HGF gene transfer and provide an overview of ideal treatment regimes for various nervous injuries and disorders. PMID- 20021364 TI - Are circulating monocytes as microglia orthologues appropriate biomarker targets for neuronal diseases? AB - Microglial cells, in contrast to other central nervous system cell types such as neurons and macroglia, are of myeloid origin. They constitute the immune cells of the brain and are involved in neuroinflammatory and neurodegenerative processes. Moreover, diseases of the central nervous system with an inflammatory component are characterized by the migration of bone marrow-derived monocytes into the brain where they differentiate into microglia, the "tissue macrophages" of the nervous system, bearing a therapeutic potential for certain diseases by transplantation of bone marrow-derived hematopoietic stem and progenitor cells. Due to their common origin, microglial cells and monocytes/macrophages share expression of many surface receptors and signalling proteins. Moreover, there is overlap in the expression of many genes related to Alzheimer s disease. Activation of resident and blood-derived microglia in diseases of the central nervous system can be both beneficial, e.g. by degradation of protein aggregates, and detrimental, e.g. by secretion of neurotoxic factors. This review summarizes the current knowledge about the role of microglia in neurodegenerative diseases with a focus on Alzheimer s disease. Moreover, we present data how neuroinflammation is reflected by cellular changes in peripheral blood enabling the use of blood monocytes/macrophages for diagnosis, therapeutic target finding and outcome monitoring of neurodegenerative disorders. In summary, blood monocytes as microglia orthologues are an important model system to study the role of microglia in the pathogenesis of neurodegenerative diseases. They are suitable biomarker targets for diagnosis and prognosis and maybe also therapy of central nervous system disease. PMID- 20021366 TI - A new journal with an integrated approach in the study of aging and longevity. PMID- 20021365 TI - Hormones in the mentally disturbed brain: steroids and peptides in the development and treatment of psychopathology. AB - One of the more fascinating recent discoveries in neuroscience is the widespread influence of hormones on brain regions and functions underlying pathological behaviors. A story is unfolding that points to critical roles played by hypothalamic - pituitary - gonadal (HPG) and hypothalamic - pituitary - adrenal (HPA) axes on a startling array of mental disorders, from depression to dementia. The influence of peptides and steroids does not end with hormones released from the two axes, however. It is now clear that the brain has adapted, "highjacked" is more descriptive, HPG and HPA hormones for uses unrelated to their original functions in reproduction and responses to stress. Findings of neuromodulatory effects of HPA and HPG hormones on monoamine, GABA, glutamate and opioid pathways and of hormone receptors and enzymes involved in hormone synthesis, particularly of steroids, in the hippocampus, amygdala and other subcortical brain regions provide the brain with multiple evolutionary means to adapt to new functions. The complexity of the metabolic cascade for the steroids also leaves open mechanisms by which endogenous errors and exogenous chemicals could be involved in the etiology of psychopathologies. The planned review will examine the recent literature for evidence of steroidal and peptidergic influences on basic biological functions and on mood disorders, anxiety and PTSD, schizophrenia, substance abuse and dementia. Emphasis will be placed on animal models, although findings with patient populations will be prominently included. Special attention will be paid to novel pathways by which the precursors and metabolites of sex steroids can influence psychopathologies. We also will speculate on promising treatments with hormone modulators that may be useful in mollifying the symptomology of the mental disorders. PMID- 20021367 TI - Towards an understanding of the anti-aging mechanism of caloric restriction. AB - Accumulation of oxidatively altered cell components may play a role in the age related cell deterioration and associated diseases. Caloric restriction is the most robust anti-aging intervention that extends lifespan and retards the appearance of age-associated diseases. Autophagy is a highly conserved cell repair process in which the cytoplasm, including excess or aberrant organelles, is sequestered into double-membrane vesicles and delivered to the degradative vacuoles. Autophagy has an essential role in adaptation to fasting and changing environmental conditions. Several pieces of evidence show that autophagy may be an essential part in the anti-aging mechanism of caloric restriction: 1. The function of autophagy declines with increasing age; 2. The temporal pattern of the decline parallels the changes in biomarkers of membrane aging and in amino acid and hormone signalling. 3. These age-dependent changes in autophagy are prevented by calorie restriction. 4. The prevention of the changes in autophagy and biomarkers of aging co-varies with the effects of calorie restriction on life span. 5. A long-lasting inhibition of autophagy accelerates the process of aging. 6. A long-lasting stimulation of autophagy retards the process of aging in rats. 7. Stimulation of autophagy may rescue older cells from accumulation of altered mtDNA. 8. Stimulation of autophagy counteracts the age-related hypercholesterolemia in rodents. It is suggested that the pharmacological intensification of suppression of aging (P.I.S.A. treatment) by the stimulation of autophagy might prove to be a big step towards retardation of aging and prevention of age-associated diseases in humans. PMID- 20021368 TI - The mitochondrial free radical theory of aging: a critical view. AB - The Mitochondrial Free Radical Theory of Aging (MFRTA) proposes that mitochondrial free radicals, produced as by-products during normal metabolism, cause oxidative damage. According to MFRTA, the accumulation of this oxidative damage is the main driving force in the aging process. Although widely accepted, this theory remains unproven, because the evidence supporting it is largely correlative. For example, long-lived animals produce fewer free radicals and have lower oxidative damage levels in their tissues. However, this does not prove that free radical generation determines life span. In fact, the longest-living rodent Heterocephalus glaber- produces high levels of free radicals and has significant oxidative damage levels in proteins, lipids and DNA. At its most orthodox MFRTA proposes that these free radicals damage mitochondrial DNA (mtDNA) and in turn provoke mutations that alter mitochondrial function (e.g. ATP production). According to this, oxidative damage to mtDNA negatively correlates with maximum life span in mammals. However, in contrast to MFRTA predictions, high levels of oxidative damage in mtDNA do not decrease longevity in mice. Moreover, mice with alterations in polymerase gamma (the mitochondrial DNA polymerase) accumulate 500 times higher levels of point mutations in mtDNA without suffering from accelerated aging. Dietary restriction (DR) is the only non-genetic treatment that clearly increases mean and maximum life span. According to MFRTA caloric restricted animals produce fewer mitochondrial reactive oxygen species (mtROS). However, DR alters more than free radical production (e.g. it decreases insulin signalling) and therefore the increase in longevity cannot be exclusively attributed to a decrease in mtROS generation. Thus, moderate exercise produces similar changes in free radical production and oxidative damage without increasing maximum life span. In summary, available data concerning the role of free radicals in longevity control are contradictory, and do not prove MFRTA. In fact, the only way to test this theory is by specifically decreasing mitochondrial free radical production without altering other physiological parameters (e.g. insulin signalling). If MFRTA is true animals producing fewer mtROS must have the ability to live much longer than their experimental controls. PMID- 20021369 TI - Stress, aging and reliability of antioxidant enzyme defense. AB - Clinical and experimental data point to existence of disturbances of adaptive ability of aged organism to extreme impacts. However mechanisms of these disturbances are not clear yet. The purpose of the investigation was to study age related changes in reaction of erythrocyte antioxidant enzyme system in response to acute psycho-emotional stress and a possible role of these changes in age related alterations of oxygen blood transport in nonhuman primates. Ten young (6 8 years) and ten old (20-26 years) healthy female rhesus monkeys were subjected to acute moderate psycho-emotional stress (two hours squeeze cage restraint) at 1500h. Plasma cortisol, lipid peroxidation products (TBARS) and activities of superoxide dismutase (SOD), glutathione peroxidase, gluthatione reductase (GR), and gluthatione-S-transferase in erythrocytes were measured before stress and at 30, 60, 120, 240 min and 24 hours after beginning of the stress. We have found for the first time that SOD activity decreased in response to the stress in young monkeys while it increased in the half of old monkeys. Young animals also demonstrated essentially higher increase in GR activity and plasma cortisol level in response to the restraint in comparison with old monkeys. Level of TBARS did not practically change in response to the stress in young animals and significantly increased in old monkeys. The study demonstrated that the age related alterations in SOD and GR stress responsiveness lead to activation of peroxide oxidation of lipids that may be considered as an important factor of aging damage of erythrocyte functioning and reliability of oxygen transport to tissues under stress conditions. PMID- 20021370 TI - Inflammation in neurodegenerative disorders: friend or foe? AB - Inflammation plays a role in the development of Alzheimer's disease (AD). Several cytokines and chemokines have been detected both immunohistochemically and in cerebrospinal fluid from patients. Some of them, including Tumor Necrosis Factor alpha, Interferon-gamma-inducible Protein-10, Monocyte Chemotactic Protein-1 and Interleukin-8, are increased in AD and in Mild Cognitive Impairment (MCI), considered the prodromal stage of AD, suggesting that these modifications occur very early during the development of the disease, possibly explaining the failure of trials with anti-inflammatory agents in patients with severe AD. Further evidence suggests that cytokines and chemokines could have a role in other neurodegenerative disorders, such as Frontotemporal Lobar Degeneration and Amyothrophic Lateral Sclerosis. In this regard, analogies and differences among these neurodegenerative disorders will be discussed. Neurodegenerative disorders are considered multifactorial diseases, and genetic factors influence pathological events and contribute to change the disease phenotype from patient to patient. Gene polymorphisms in crucial molecules, including cytokines, chemokines and molecules related to oxidative stress, may act as susceptibility factors, increasing the risk of disease development, or may operate as regulatory factors, modulating the severity of pathogenic processes or the response to drug treatment. With these premises, genetic studies recently carried out will be described and discussed in detail. PMID- 20021371 TI - Hypopituitarism in the elderly: multifaceted clinical and biochemical presentation. AB - Hypopituitarism (HYPO) is a rare and under-investigated pathology in the elderly. AIM: to review our case records of patients > or =65 yrs with first diagnosis of anterior global hypopituitarism, in order to evaluate presentation symptoms, etiology, biochemical and hormonal pictures, pituitary morphology, and efficacy of therapy. PATIENTS: 15 patients (65-82 yrs) were studied: in 11 (73%) HYPO was secondary to pituitary macroadenoma (non-secreting in 10 and GH-secreting in 1); in 3 it was associated to empty sella, and in 1 to pituitary hypoplasia. RESULTS: major presenting symptoms were visual-field defects and asthenia (40%) but also memory and/or gait impairment and nausea (30%) and depression (20%) were significantly observed. Dyslipidemia (73%), anemia (20%) and severe hyponatremia (13%) were found. After starting substitutive therapy and clinical improvement, 10 patients with macroadenoma underwent uneventful neurosurgery, which improved visual alterations but not pituitary function. Immunohistochemistry showed positivity for FSH in one patient and for GH in one patient. Six out of the eight patients with a post-surgical tumor remnant required treatment (surgery/radiotherapy/somatostatin analogue treatment in the acromegalic patient). CONCLUSIONS: The diagnosis of HYPO is often delayed in the elderly, since symptoms may be ascribed to aging and associated comorbidities. In our series, most of the aspecific symptoms were retrospectively addressed to HYPO since their resolution/improvement with replacement therapy. The prevalent cause of HYPO remains non-functioning pituitary macroadenomas. Hyponatremia can be a life-threatening presenting symptom. Symptoms considered apparently aspecific in the elderly should be investigated in order to possibly diagnose an important treatable disorder as HYPO. PMID- 20021372 TI - Correlation between ROS production and InsP3 released by granulocytes from type 1 diabetic patients in a cAMP-dependent manner. AB - BACKGROUND: Diabetes is associated with a pro-inflammatory status characterized by an increased production of inflammatory molecules. Reactive oxygen species (ROS) and cAMP elevating agents represent two molecular systems, normally generated during inflammation. These molecules could be responsible for the alteration of signaling pathways. In the present paper we have studied the correlation between ROS generation and inositolpolyphosphates (InsP(1), InsP(2) InsP(3) and InsP(4)) released by granulocytes from Type 1 diabetic patients (DM1) in the presence or in the absence of cyclic AMP-elevating agents. METHODS: The effect of cAMP on ROS production was quantified in a chemoluminescence assay luminol-dependent (RLU/min). InsP(1), InsP(2) InsP(3) and InsP(4) were quantified by inositol-H(3) in a Beta-counter and the results were expressed as count per minute (CPM). RESULTS: The elevation of intracellular level of cAMP inhibited both InsP(3) and ROS production in granulocytes from healthy subjects and activated in the cells from Type 1 diabetic patients. InsP(1), InsP(2) and InsP(4) did not show significant alteration in both studied cells. There was a significant correlation between InsP(3) and ROS in the presence of elevated content of cAMP. This correlation was observed in a 15 minutes reaction for healthy subjects and in 120 minutes for DM1. CONCLUSIONS: The importance of both InsP(3) release and ROS production in an inflammatory process and tissue pathophysiology in Type 1 diabetic patients is still under debate because hyperglycemia accelerates generation of oxidative stress and may play an important role in the development of complications in diabetes. Thus, our results demonstrated alteration in metabolic response in granulocytes from Type 1 diabetic patients and it may be important for the development of therapeutic processes and drugs that interfere with signaling of ROS generation and may contribute to the improvement of the severe complications of diabetes. PMID- 20021373 TI - Tau as a molecular marker of development, aging and neurodegenerative disorders. AB - The purpose of this work is to review the changes that take place in the microtubule associated protein tau during neuronal development, aging and neurodegeneration. Human tau protein is expressed from a single gene located on chromosome 17. The DNA is transcribed into nuclear RNA and this RNA, by alternative splicing, yields different mRNA species which are developmentally regulated. In aging, or in neurodegenerative disorders, post translational modifications of tau, such as phosphorylation, could take place, and new tau isoforms may appear. Thus, tau isoforms can be used as markers to follow neuronal development, aging or neurodegeneration. PMID- 20021374 TI - High velocity power training in older adults. AB - Increases in both the age and the number of older adults in the United States will likely result in more people living with functional limitations and physical disabilities. The impact of this change in demographics will not only significantly impact older adult quality of life but may overwhelm existing health care services for this population. Resistance training with a strengthening component is currently recommended for older adults who wish to increase strength and overall health. However, muscle power has recently been found to contribute more to improvement in physical functioning than muscle strength and is becoming a focus of many resistance training studies in older adults. This review will discuss the current research supporting the implementation of traditional strength-enhancing resistance training, examine the contribution of muscle power to function, explore the rationale for implementing high velocity power training interventions, and review the recent literature on these novel power training interventions in older men and women. Recommendations for future research will be discussed. PMID- 20021375 TI - A Tai Chi Chuan training model to improve balance control in older adults. AB - The first goal of this article is to present nine Tai Chi Chuan training principles and incorporate them into a current model of motor control and motor learning theory. The second goal is to present a Tai Chi Chuan training model. The third goal is to construct a theory as to how Tai Chi Chuan principles may improve balance and motor skills in an aging population. Evidence from the areas of motor control, biomechanics, and human physiology are drawn upon to build a theory of motor skill learning and construct a Tai Chi Chuan training model. PMID- 20021376 TI - Extending life using tissue and organ replacement. AB - There is a critical shortage of organs available for donation to patients suffering from degenerative diseases of various organ systems. This shortage becomes more severe yearly, as the aging population grows and such diseases become more common. The fields of regenerative medicine and tissue engineering now offer hope for these aging patients with new advances in material science, cell transplantation, and bioengineering. Novel methods and materials to construct biological substitutes for diseased and injured tissues are being developed, and the goal is to restore and maintain normal function to the patient. The field of stem cell research is rapidly advancing, offering unforeseen options for treatment. For example, therapeutic cloning, where an enucleated oocyte receives a donor cell nucleus, yielding pluripotent stem cells, offers a potentially limitless source of cells for tissue engineering applications. However, recent discoveries in this field indicate that the use of non-controversial cells and tissues also has enormous potential for tissue engineering purposes. This article gives an overview of recent advances in regenerative medicine and describes their applications in tissue and organ replacement technology, as well as how these technologies offer new therapies to a person facing end-stage organ failure. There is a growing optimism that the successes in this field will achieve the goal of significantly extending the life of the patient. PMID- 20021377 TI - Dietary habits of Mongolian people, and their influence on lifestyle-related diseases and early aging. AB - Mongolians are known to have relatively short life expectancy. In order to examine the role of dietary habits in the early aging of Mongolians, the food intake inquiry, anthropometric measurements and blood clinical tests were performed for 365 healthy inhabitants in Murun, a northern Mongolia city, and compared to those of Japanese. Murun inhibitants were found to have a characteristic dietary habit of taking large amounts of meat, milk, dairy products and wheat flour products, in contrast little vegetables, fruits and fishes. The daily calorie intake of the adults was estimated to be 2,525 kcal, and the fat/total calorie ratio was calculated 33.7%, about 1.3-fold higher than that of Japanese. The intake ratio of fatty acid from the Mongolian foods, saturated : mono-unsaturated : poly-unsaturated fatty acids (PUFA) ratio, was 10.3 : 7.8 : 3.0. Results of blood clinical tests showed significantly higher levels of serum triglycerides, low-density lipoprotein cholesterol (LDL) and homocysteine, and lower levels of high-density lipoprotein cholesterol (HDL), n-3 PUFA, folic acid and adiponectin, in comparison with those of Japanese. In addition, the Mongolians were also found to have significantly high levels of oxidative stress markers, such as serum malondialdehyde-modified LDL (MDA-LDL), urinary 8-hydroxy-2'-deoxyguanosine (8-OHdG) and serum reactive oxygen metabolites (ROM). The serum ROM level in the Mongolians seemed to associate with their body fat ratio (p<0.05), and was significantly inverse-correlated to handgrip strength (p<0.001). Obesity was observed at a high incidence in the subjects over 30-year old, and over 40-year old their handgrip ability was markedly decreased. These findings suggest that in the Mongolians the dietary habits associate with their lifestyle-related diseases and early aging, and the improvement of dietary habits is an effective strategy for health promotion of the inhabitants. PMID- 20021378 TI - Effects of aging on thyroidal function and proliferation. AB - The endocrine system is as affected by aging as are other systems. The effect of aging on the hypothalamus-pituitary-thyroid function is still controversial. Human aging was reported as associated with a decrease in thyrotropin (TSH) secretion, but increased TSH levels in relatively healthy elders are also reported. The main point discussed is whether this increase in the immunoreactive TSH of aged subjects, and related changes in thyroid function, are "physiologic" consequences of aging on the hypothalamus-pituitary-thyroid axis or are induced by non-thyroid illnesses and/or drug use, frequent in the elderly. There are strong evidences of decreased hypothalamus-pituitary-thyroid axis activity as well as decreased thyroxine metabolism (5'-deiodination) in humans, and other mammals. For now, we must consider that the hypothalamus-pituitary-thyroid axis is affected at all three levels by normal aging, and the mild state of "total" hypothyroidism during aging is completed by a reduced response of target cells/tissues to thyroid hormones. Despite the decreased response of the old rat thyroid to TSH there is no decrease in the glands mass. Ras proteins are involved in the transduction of growth factor signals by surface receptors, in thyroid as well as in other tissues, and are key components of downstream signaling through several pathways. Ras activation of Raf, and of extracellular-signal-regulated kinases (ERK) is an important signaling pathway for many Ras effects. Very little is known about the modulation of Ras expression in the aging thyroid. We detected an increase in Ras expression in thyroids of old rats, but the signal transduction by pERK was decreased, suggesting that another RAS-signaling pathway could be activated and responsible for the maintenance of the thyroid volume. PMID- 20021379 TI - Intraventricular implant of encapsulated CNTF-secreting fibroblasts ameliorates motor deficits in aged rats. AB - The impact of intraventricular ciliary neurotrophic factor (CNTF) on motor function in aged rats was evaluated. Spontaneous locomotion and motor coordination were quantified in young (5-6 months) and aged (24-25 months) rats. Relative to young animals, aged rats were significantly less active, fell more rapidly from a rotating rod, and were unable to maintain their balance on a wooden beam. Aged animals received bilateral intraventricular implants of polymer encapsulated fibroblasts that were genetically modified to secrete CNTF. Controls received either no implant or capsules loaded with mock transfected cells. One month after implantation the aged animals that received CNTF implants were significantly more active and were improved on the rotorod and beam balance tests. The improvement in performance on the rotorod and beam balance tests was dependant on the task difficulty and dissipated at higher rotations (rotorod) and smaller beam widths (beam balance). No recovery was seen in aged animals receiving control implants. Postmortem removal of the encapsulated cells confirmed that they continued to secrete CNTF. These data are the first to suggest that intracerebral delivery of CNTF might be useful for slowing or reversing age-related changes in motor function. PMID- 20021380 TI - Inflammation and age-related iron accumulation in F344 rats. AB - Evidence suggests chronic inflammation and iron accumulation may play a role in the pathogenesis of Parkinson's disease (PD) as inflammation and iron levels increase with age and appear in the disease pathology. It is hypothesized that an aggravated inflammatory response and iron accumulation, as a function of age, increase oxidative stress and participate in the pathogenesis of PD. Intracranial injection of the bacterial endotoxin, lipopolysaccharide (LPS), has been shown to induce microglia activation, oxidative stress, mitochondrial impairment, iron accumulation, and dopaminergic neurodegeneration within the substantia nigra. We tested the hypothesis that injection of LPS into the striatum would increase iron accumulation in the substantia nigra of aged rats compared to young ones. Our results showed that four weeks post injection, LPS significantly increased microglia activation, lipid peroxidation, ferritin expression, and total nigral iron content in aged rats. In addition, LPS significantly altered the turnover ratio of homovanillic acid to dopamine. Thus, an age-related increase in iron as well as susceptibility to inflammation may play an important role in PD-related neurodegeneration, as free radicals produced from the inflammatory response can become more toxic through increased ferrous iron catalyzed Fenton chemistry. This may enhance oxidative stress, exacerbate microglia activation, and drive the progression of PD. PMID- 20021381 TI - The interface between Alzheimer's disease, normal aging, and related disorders. AB - Since the earliest descriptions of Alzheimer's disease (AD), the presence of senile plaques (SP) and neurofibrillary tangles (NFT) have been regarded as the typical pathological hallmarks of the disease. Studies over the last twenty years, however, have reported a considerable degree of heterogeneity within the AD phenotype and as a consequence, an overlap between the pathological features of AD not only with normal aging, but also with disorders related to AD. This review discusses: 1) the degree of heterogeneity within AD, 2) the concept of an 'interface' between disorders, 3) the nature and degree of the interface between AD and normal aging, vascular dementia (VD), the tauopathies, synucleinopathies, and prion disease, and 4) whether the original status of AD should be retained or whether AD, normal aging, and the related disorders should be regarded as representing a 'continuum' of neuropathological change. PMID- 20021382 TI - Mood, memory and movement: an age-related neurodegenerative complex? AB - The following review was constructed as a concept paper based on a recent workshop on neurodegenerative disease sponsored by the National Institute on Aging (NIA), the American Geriatric Society (AGS), and the John A. Hartford Foundation. The meeting was entitled "Thinking, moving and feeling: Common underlying mechanisms? 4(th) Annual Bedside-to-Bench Conference" and had the purpose to connect current basic and clinical findings on common brain-related alterations occurring with aging such as depression, movement disorders, and cognitive decline. Many prominent researchers expressed their opinion on aging and it was revealed that age-related brain dysfunction of any kind seems to share several risk factors and/or pathways. But can something be done to actively achieve "successful aging"? In this review, based largely on the workshop and current literature, we have summarized some of the current theories for depression, movement and cognitive impairment with aging, as well as potential preventive measures. We have also summarized the emerging need for relevant animal models and how these could be developed and utilized. PMID- 20021383 TI - An observational study of the needs and quality of life amongst patients in the treatment of Alzheimer's dementia with cholinesterase inhibitors. AB - Alzheimer's dementia is one of the most commost mental health disorders associated with aging. In its earlier stages sufferers live independently but gradually rely increasingly on loved ones or formal carers for support as the illness progresses. Its treatment involves both medical and social care. This study assesses the impact of outpatients' care and cholinesterase inhibitors in patients being treated for Alzheimer's dementia. The needs and quality of life of patients attending an outpatients dementia care service were assessed using the Camberwell Assessment of Need for the Elderly (CANE) and Quality of Life in Alzheimer's Disease: Patient and Caregiver report. Other tools used were the Problems Checklist and Carer Strain, the Minimental State Examination (MMSE) and a proforma to obtain sociodemographic details. All patients who had informal care were assessed using the questionnaires. 104 patients were seen of whom 34 were new and 70 were follow-up patients. 43 patients lived alone while the rest lived with their spouses or other relatives such as children. There was reduction in the number of CANE unmet needs and increased combined Quality of Life in Alzheimer's Dementia scores in the first three months amongst the newly referred patients. The findings suggest that outpatients' dementia care and prescribing of cholinesterase inhibitors helped to meet the needs of patients and improve patients' quality of life in first three months. PMID- 20021385 TI - Metabolic effects of resveratrol in mammals--a link between improved insulin action and aging. AB - Resveratrol, a polyphenol found in several vegetal sources, has been shown to possess lifespan-promoting properties in yeast and metazoans, including small mammals. While in yeast and low metazoans resveratrol acts mainly by activating the histone deacetylase Sir2, in mammals it appears to target - besides the Sir2 homolog SIRT1 - several crucial pathways for the control of metabolism, including the AMPK and the insulin-IGF1 receptors axis. The action of resveratrol on these pathways has been linked to its capability to i) prolong lifespan following chronic administration to mice and ii) protect from the development of diet induced obesity and obesity-dependent metabolic disorders. Here we summarise the current understanding on how resveratrol displays its remarkable properties by acting on the control of insulin secretion and by modulation of insulin action in pheripheral insulin-responsive tissues. Since resveratrol has the potential for pharmacological exploitation to prevent the establishment of insulin-resistance and thus postpone - or even prevent - the onset of type 2 diabetes, toxicologic and pharmacodynamics studies in humans have been initiated. These studies show that resveratrol is non-toxic and easily absorbed by humans. As a drawback, its bioavailability is very limited due to the fast metabolic alterations to which it is subjected in the plasma. Therefore, we also review here the efforts that have been made - in the drug discovery field - to identify new molecules endowed with resveratrol-like pharmacological properties but with better bioavailability, which could prove to possess therapeutic potential. PMID- 20021386 TI - Melatonin causes gene expression in aged animals to respond to inflammatory stimuli in a manner differing from that of young animals. AB - Groups of younger and aged mice were fed either minimal basal diet or the same diet containing 40 ppm melatonin. After 9.3 weeks half of each of these 4 groups of animals received either an intraperitoneal injection of lipopolysaccharide (LPS) or of saline. Three hours after this treatment, all animals were killed and mRNA from brains extracted. Quantitative PCR was performed on 13 selected mRNA species reflecting various aspects of the inflammatory pathway, the melatonin receptor, and a key glycolytic enzyme. An overall trend observed was that the effect of melatonin in modulating LPS-provoked immune responses differed markedly in old and young animals. Melatonin tended to enhance the reaction of younger animals to LPS but suppressed the inflammatory response of older mice. This difference with aging suggests that key immune processes are markedly altered by aging. It is likely that the ability of the immune system to mount a defense is impaired in older animals. PMID- 20021387 TI - Is the yeast a relevant model for aging of multicellular organisms? An insight from the total lifespan of Saccharomyces cerevisiae. AB - The applicability of the free radical theory of aging to the yeast S. cerevisiae is a matter of debate. In order to get an insight into this question, we studied the reproductive potential (the number of buds produced), reproductive lifespan (the time during which a yeast cell is able to divide), postreproductive lifespan (duration of life of yeast cells which ceased to divide) and total lifespan (sum of reproductive lifespan and postreproductive lifespan) of three isogenic pairs of yeast strains. Each pair contained a parent strain and a disruptant of gene(s) coding for important antioxidant enzyme(s) (CuZn-superoxide dismutase, all five peroxiredoxins or glutaredoxin 5). Although the reproductive potential was decreased in all antioxidant enzyme-deficient mutants, the differences in the reproductive lifespan between the parent strains and the mutants were less pronounced while postreproductive lifespan and total lifespan were not diminished in the mutants. These results suggest that either the free-radical theory of aging is not applicable to S. cerevisiae or that this yeast is not a proper model organism for the study of aging of higher organisms. In our opinion the latter possibility is more apparent and the increase in cell volume (unavoidable for a cell propagating by budding) rather than accumulation of oxidative damage may be the main reason for the cessation of budding (and perhaps postreproductive death) in S. cerevisiae. PMID- 20021388 TI - Possible links of age related hypertension and evolution imposed features of heart and aorta. AB - The left ventricle thickness is a limiting factor of optimal heart size and strength. Due to disappearance of all the features compromising left ventricular compliance, mammalian heart has decreased vascular density and coronary vessel diameter and it requires sufficient diastolic aortic pressure for the left ventricle perfusion. Atrial muscle and the right ventricle are perfused during the entire heart cycle. The systolic pressure in the left ventricle forces blood vessels in the muscle wall to collapse, particularly in the subendocardial muscle layer. This makes the most active part of the heart prone to hypoxia. Optimal perfusion of the left ventricle wall requires sufficient aortic pressure during diastole, making individuals with higher diastolic pressures advantageous, in situations requiring combination of increased heart rate and output. Described mechanisms might have contributed to the hereditary quality of age-related hypertension in humans. PMID- 20021389 TI - Nanoparticulated quercetin in combating age related cerebral oxidative injury. AB - Reactive oxygen species e.g. O(2)(*-), H(2)O(2) and *OH generated by the induction of oxidative stress exert a potential threat on the activity of endogenous antioxidant enzymes and substantially influence the aging process and age-dependant neuropathology. Chemical antioxidant is almost ineffective in protecting neuronal cells from oxidative damage as Blood Brain Barrier exists in between blood and brain interstitial fluid that restricts undegradable influx from the circulation into cerebral region. Quercetin (QC), a flavonoidal antioxidant is known as a potent antioxidant for its polyphenolic configuration. Formulation of QC in polylactide nanocapsule has been done and the efficacy of this vesicular flavonoid has been tested against cerebral ischemia induced oxidative damage in young and old rat brains. Antioxidant potential of QC loaded in nanocapsule (QC 7.2 mmol/kg b.wt., size 50 nm) was investigated by an in vivo model of cerebral ischemia and reperfusion on Sprague Dawley young (2 months, b.wt. 160-180 g) and aged (20 months, b.wt. 415-440 g) rats. Diene level, the index of lipid peroxidation and GSSG/GSH ratio were found to be higher in normal aged, compared to normal young rat brain. Endogenous antioxidants activities were lower in aged rat brain compared to young. Further reduction of these antioxidants were observed in aged rat brain by the induction of cerebral ischemia - reperfusion. Nanocapsule encapsulated QC treatment resulted a significant protection to endogenous antioxidant enzymes against ischemia induced oxidative damage in neuronal cells of young and old rats. PMID- 20021391 TI - Human skeletal muscle aging and the oxidative system: cellular events. AB - As we age, the aerobic and functional capacities of our major physiological systems progressively decline. In the case of the neuromuscular system, reductions in strength and mobility cause a deterioration in motor performance and in turn a greater tendency to fall (with increased risk of fractures), impaired mobility, disability and loss of independence in the elderly. Given the increase in our life expectancy and the consequent growth in the elderly population, these conditions will have an increasing impact on modern healthcare systems, and their prevention and attenuation needs to be addressed. Several intervention strategies have been used to improve motor performance among the aging. At the cellular level, aging is caused by a progressive decline in mitochondrial function that results in the accumulation of reactive oxygen species (ROS) generated by the addition of a single electron to the oxygen molecule As the level of oxidative stress in skeletal muscle increases with age, the production of some antioxidant enzymes increases adaptively to compensate in part. The aging process is characterized by an imbalance between an increase in the production of reactive oxygen species in the organism and the antioxidant defences as a whole. The goal of this review is to examine the results of existing studies on oxidative stress in aging human skeletal muscles, taking into account different physiological factors (sex, fiber composition, muscle type and function). PMID- 20021390 TI - Thyroid hormone levels in the prefrontal cortex of post-mortem brains of Alzheimer's disease patients. AB - Converging evidence suggests a possible link between thyroid state and Alzheimer's disease (AD), including a higher probability of dementia in individuals with higher TSH levels and a two-fold risk of AD in patients with hypothyroidism. Thyroid hormones modulate factors associated with AD, including amyloid precursor protein expression in the brain, suggesting a possible role for thyroid hormone in AD pathology. The present study is the first to directly evaluate brain thyroid hormone levels in AD. Triiodothyronine (T(3)) and thyroxine (T(4)) levels were measured with radioimmunoassay (RIA) in post-mortem samples of prefrontal cortex of patients with pathologically confirmed AD, including Braak stage I-II (n=8), Braak stage V-VI (n=8), and controls without any primary neurological disease (n=8). T(4) levels did not differ between groups. T(3) levels were significantly lower in Braak stage V-VI brains relative to controls, but there was no statistically significant difference between T(3) levels in Braak stage I-II versus controls. Results suggest that the conversion of T(4) to T(3) may be affected in advanced AD, perhaps due to alterations in deiodinase activity. Reduced conversion of T(4) to T(3) in AD may be associated with both AD pathology and the clinical presentation of dementia. PMID- 20021392 TI - Interleukin-4-induced oxidative stress via microglial NADPH oxidase contributes to the death of hippocampal neurons in vivo. AB - We investigated the effects of interleukin-4 (IL-4), a well-known anti inflammatory cytokine, on thrombin-treated rat hippocampi in vivo. Intrahippocampal injection of thrombin resulted in a significant loss of hippocampal CA1 neurons, as determined by Nissl staining and NeuN immunohistochemistry. Thrombin-induced neurotoxicity was accompanied by substantial microglial activation, as demonstrated by OX-42 immunohistochemistry. In parallel, Western blot analysis and hydroethidine histochemistry revealed activation of NADPH oxidase (as demonstrated by increased translocation of the cytosolic proteins p67(phox) and p47(phox)), generation of reactive oxygen species (ROS), and oxidative damage in the hippocampal CA1 area, where degeneration of hippocampal neurons was evident. Interestingly, immunohistochemical and biochemical analysis demonstrated that intrahippocampal injection of thrombin increased immunoreactivity and levels of IL-4 as early as 8 h post-treatment, reaching a peak at 7 days that was maintained for up to 14 days. Moreover, double-label immunohistochemistry detected IL-4 immunoreactivity solely in activated microglia. In experiments to explore the involvement of IL-4 in neurotoxicity, IL-4-neutralizing antibodies significantly increased the survival of CA1 hippocampal neurons at 7 days post-thrombin treatment. Consistent with these results, IL-4 neutralization inhibited activation of NADPH oxidase, ROS production and oxidative damage. Thus, the present study is the first to demonstrate that IL-4 generates microglial NADPH oxidase-derived oxidative stress and leads to the degeneration of hippocampal neurons in vivo, as occurs in Alzheimer's disease. PMID- 20021393 TI - Prematurely aged children: molecular alterations leading to Hutchinson-Gilford progeria and Werner syndromes. AB - Ageing is thought to be a polygenic and stochastic process in which multiple mechanisms operate at the same time. At the level of the individual organism ageing is associated with a progressive deterioration of health and quality of life, sharing common features such as: alopecia and grey hair, loss of audition, macular degeneration, neurodegeneration, cardiovascular diseases, osteoporosis, cataract formation, type-2 diabetes, lipodystrophies; a generally increased susceptibility to infection, autoimmune disorders and diseases such as cancer; and an impaired ability to cope with stress. Recent studies of mechanisms involved in the ageing process are contributing to the identification of genes involved in longevity. Monogenic heritable disorders causing premature ageing, and animal models have contributed to the understanding of some of the characteristic organism-level features associated with human ageing. Werner syndrome and Hutchinson-Gilford progeria syndrome are the best characterized human disorders. Werner syndrome patients have a median life expectancy of 47 years with clinical conditions from the second decade of life. Hutchinson-Gilford progeria syndrome patients die at a median age of 11-13 years with clinical conditions appearing soon after birth. In both syndromes, alterations in specific genes have been identified, with mutations in the WRN and LMNA genes respectively being the most closely associated with each syndrome. Results from molecular studies strongly suggest an increase in DNA damage and cell senescence as the underlying mechanism of pathological premature ageing in these two human syndromes. The same general mechanism has also been observed in human cells undergoing the normal ageing process. In the present article the molecular mechanisms currently proposed for explaining these two syndromes, which may also partly explain the normal ageing process, are reviewed. PMID- 20021394 TI - Progress and vision for 'Current Aging Science'. PMID- 20021395 TI - Does aging stop? AB - Human mortality data show stabilization in mortality rates at very late ages. But human mortality data are difficult to interpret because they are affected by changing medical practices and other historically variable causes of death. However, in the 1990s, data from a variety of labs showed that the mortality rates of medflies, fruit flies, wasps, yeasts, and nematodes also stabilize at very late ages. These reproducible "mortality-rate plateaus" forced biologists to develop theories for their existence. There are two main theories of this kind. "Lifelong heterogeneity" theories suppose that highly robust subcohorts are more abundant at later ages because less robust subcohorts have mostly died off. On this type of theory, aging does not stop; aging continues inexorably in all individuals. In contrast, in evolutionary theories for mortality-rate plateaus, based on the eventual plateaus in Hamilton's Forces of Natural Selection at late ages, aging does indeed stop. A variety of experiments have cast doubt on lifelong heterogeneity theories as explanations of mortality-rate plateaus. A few experiments have corroborated the Hamiltonian theory. This has the important corollary that it appears to be possible for aging to stop, at sufficiently late ages, at least among some populations. The implications of this result for aging research are profound. Most importantly, it suggests the possibility that the physiology of adults undergoing aging may be substantially different from the physiology of life after aging. PMID- 20021396 TI - Mitochondria, cellular stress resistance, somatic cell depletion and lifespan. AB - The causes of aging and determinants of maximum lifespan in animal species are multifaceted and complex. However, a wealth of experimental data suggests that mitochondria are involved both in the aging process and in regulating lifespan. Here we outline a somatic cell depletion (SCD) model to account for correlations between: (1) mitochondrial reactive oxygen species and lifespan; (2) mitochondrial antioxidant enzymes and lifespan; (3) mitochondrial DNA mutation and lifespan and (4) cellular stress resistance and lifespan. We examine the available data from within the framework of the SCD model, in which mitochondrial dysfunction leading to cell death and gradual loss of essential somatic cells eventually contributes to the decline in physiological performance that limits lifespan. This model is useful in explaining many of the mitochondrial manipulations that alter maximum lifespan in a variety of animal species; however, there are a number of caveats and critical experiments outstanding, and these are outlined in this review. PMID- 20021397 TI - The association of very high hair manganese accumulation and high oxidative stress in Mongolian people. AB - Oxidative stress induces several diseases and early aging. Previously, we reported that Mongolians are exposed in high oxidative stress, which may cause their early aging. In this study, to know the reason of high oxidative stress, we measured hair metals. This investigation was performed in Murun city, in the northern area of this country, and 469 healthy subjects, ranging from 10 to 82 years of age, were randomly enrolled. Oxidative stress was evaluated by the levels of serum reactive oxygen metabolites (ROM), malondialdehyde-modified low density lipoprotein (MDA-LDL) and urinary 8-hydroxy-2'-deoxyguanosine (8-OHdG). Antioxidant capacity (AOC) was estimated by the levels of biological antioxidant potential (BAP) and superoxide dismutase (SOD) activity. Scalp hair metals were measured using an inductively coupled plasma mass spectrometry method. Murun subjects showed high ROM levels of 394+/-75 Carr U (n=342), compared with Japanese healthy subjects (n=356, 326+/-51 Carr U, p<0.001). MDA-LDL and 8-OHdG levels also showed high levels. While, BAP levels of Murun subjects were 2263+/ 203 micromol/L (n=210), Japanese subjects (n=356, 2087+/-215 micromol/L, p<0.001). SOD activities were also high, suggesting that the high oxidative may accelerate the state of AOC. Murun subjects demonstrated high accumulation of several metals in the hairs. In particular, Mn accumulation exhibited from 2 fold to 40 fold increases of Japanese standard. These findings are indicative that the high Mn accumulation may contribute to the high oxidative stress. The mechanism of its high accumulation was not explained by food materials or drinking water. We should further investigate another influence such as sandy wind. In order to suppress the high oxidative stress, elimination of the high Mn accumulation should be urgently studied. PMID- 20021398 TI - Visuospatial memory in healthy elderly, AD and MCI: a review. AB - In the literature it is commonly reported that several spatial abilities decline with normal aging, even though such a decline is not uniform. So far, it is not yet clear which spatial components present a normal age-related decline, which ones are preserved and at what point the deficit is so severe to represent an index of mild cognitive impairment (MCI) or a symptom of potential degenerative progression as in the early-stage Alzheimer's disease (AD). In particular, AD (from early onset) is characterised by impairments in constructive abilities, visuospatial intelligence, spatial short-term memory deficits, and disorders of spatial orientation (topographical disorientation). MCI indicates a condition, generally affecting older individuals, characterized by cognitive deficits including memory and/or non memory impairments and at high risk of progression to dementia. Three MCI subgroups have been distinguished and a very high risk of developing AD is associated to the amnestic MCI subtypes. Further, recent studies have suggested that the allocentric component of spatial memory might be taken as predictor of AD from MCI. Given the frequency of visuospatial deficits in early stage AD, evaluation of visuospatial processes is a promising approach to find predictive markers of AD. Here we report a review of the literature exploring specific visuospatial components in normal aging, MCI, and AD. In this way we could shed some light on the role of these components in the progression from MCI to AD and pave the way for future studies. PMID- 20021399 TI - Greater specificity of sensorimotor learning in the elderly when acquiring an interceptive task. AB - In this investigation we assessed the extent to which young and elderly individuals become dependent on the specific visual situation present during practice of an interceptive task. Young and elderly participants practiced extensively a task of intercepting a virtually moving target under full vision or visual occlusion of the last 600 ms of target displacement. Before and after practice they were assessed in four visual conditions varying the time interval of visual display. The results showed that the elderly practicing under full vision had a progressive increase of temporal errors as a function of the period of visual occlusion after task acquisition. The elderly practicing under visual occlusion, conversely, achieved improved performance only in the visual condition experienced during task acquisition. Young individuals showed greater adaptability, presenting similar performance across visual conditions. Development of specific visuomotor integration only for the elderly seems to be related with the higher status that vision holds for movement control at this age. PMID- 20021400 TI - Effect of gender on main clinical chemistry parameters in aged rats. AB - INTRODUCTION: Clinical laboratory data are crucially important for the diagnosis, prognosis and therapy of aging-related diseases. The age-dependent changes in the findings of routine clinical-chemistry analyses may influence the evaluation of health status of the elderly people. Serum endogenous antioxidant levels and lipid profiles are considered to be related to the progress of age-related diseases. Analyses of these routine parameters are also important for the evaluation in the elderly. Although the analyses of serum biochemical parameters are important, there have been not enough data on changes in the levels of parameters with aging in both genders. However, elderly people generally suffer from aging-related diseases or may be using some medication that interferes with the accuracy of the result; it is thus preferred that the same procedures be applied in aged rats as a model of human aging. METHODS: Aged Sprague-Dawley rats (24 months) of both genders were used in the current study. Serum total protein, albumin, bilirubin, uric acid, lipid profiles, iron, and total iron-binding capacity levels were determined on the same day of collection by standard clinical chemistry laboratory methods. RESULTS: Serum endogenous antioxidant parameters such as uric acid and bilirubin levels varied with gender in spite of almost no changes in the serum albumin and total iron binding capacity levels. Among the main parameters studied, lipid profile, conjugated bilirubin, and uric acid levels of male rats were significantly higher than in the aged female group. On the other hand, serum unconjugated bilirubin and iron levels were all found to be lower in the aged male group. CONCLUSION: Our findings support our conviction that serum biochemical parameters of aged rats have a controlling role in differing regulating mechanisms through gender differences. The gender-related data on these main serum clinical chemistry parameters in aged rats would be useful in studies of aging-related disorders using this model. PMID- 20021401 TI - The use of sensory nerve stimulation and compression bandaging to improve sensory nerve function and healing of chronic venous leg ulcers. AB - Low frequency transcutaneous sensory nerve stimulation (LF-SNS) [International Patent PCT/AU2004/001079: "nerve function and tissue healing" (Khalil, Z.)] improves sensory nerve function and accelerates wound healing of older animals. INTERVENTION: Double blind, placebo controlled randomised trial of LF-SNS for 5 minutes, twice daily for up to 12 weeks, on healing of chronic venous leg ulcers in older people. Four layer compression bandaging was the standard therapy. OUTCOME MEASURES (METHODS): Wound healing and the rate of epithelialisation (calculated from serial wound area estimation), microvascular blood flow (measured using LASER Doppler flowmetry), transcutaneous oxygen tension (measured using a transcutaneous oxygen monitor, TCM400 Radiometer). Sensory nerve activity (assessed via measuring electrical cutaneous perception threshold using the Neurometer((R))CPT and the flare response to 5% capsaicin (a selective activator of C fibres). PARTICIPANTS: 14 older people with chronic venous ulcers randomly allocated to active (mean age 74.8+/-2.3 years) and 15 to Sham nerve stimulation (mean age 76.5+/-2.6 years). RESULTS: Microvascular blood flow improved in all participants. A high proportion of wounds healed ( approximately 60%). There were trends for better C-fibre function and faster healing rates in the Active group (1.1+/-0.3 cm(2)/wk) compared to the Sham group (0.6+/-0.2 cm(2)/wk) but failed to reach statistical significance due to the small sample size. CONCLUSIONS: The improvement in microvascular blood flow in both groups was an unexpected finding that has not previously been described. Most likely this was due to the four layer compression bandaging provided to all participants. Improved microvascular blood flow may be a significant contributor to wound healing. The observed trends to increased healing rates and improvements in C-fibre function in the actively stimulated group compared to the sham group warrant further studies of LF-SNS as an adjunct therapy for chronic venous leg ulcers. An exploration of the possibility that LF-SNS induced-improvement in C-fibre function could protect against future ulceration is also warranted. PMID- 20021402 TI - Fat and aging: a tale of two tissues. AB - The thrifty aged hypothesis (TAH) holds that aging is a "saving program", a set of strategies all of them focused to reduce energy consumption in the post reproductive age. Aged adults remove fewer energy resources from the environment, and the remaining fuel can be profitably used by offspring. Thus, from an evolutionary point of view, the variable submitted to natural selection would be the amount of energy removed by post-reproductive individuals from the environment, favoring "reduced energy extraction" (REE) strategies. Senescence is characterized by a set of metabolic changes, and one of them is a deep modification in adipose tissue. A pattern of progressive loss of subcutaneous (SF) and gain of visceral fat (VF) throughout life has been demonstrated. The evolutionary reasons for this change have not been explored. No one has questioned why energy is deposited in VF. It appears that VF is not the best place for storing fat. The objective of this theoretical paper is to analyze the relationship between aging and adipose tissue. Furthermore, an evolutionary explanation is proposed. Fat redistribution could be thought of as a consequence of the TAH. While SF responds to the equilibrium between energy needs and fuel supply, VF acts in a dysfunctional manner. VF is not coordinated with pathways that regulate the state of fat depots. Hence, the purpose of the changes observed upon aging is to limit the effectiveness of those pathways that force the organism to obtain energy from the environment. In summary, adipose tissue modifications promote REE strategies. PMID- 20021403 TI - "Oxygen supply" as modulator of aging processes: hypoxia and hyperoxia models for aging studies. AB - Cell growth is regulated by several factors, including oxygen supply, which influence cell metabolism. Aging is characterized by decreased oxygen supply to tissue, a reduction of tissue PO(2) and of the activity of several enzymes and metabolic factors. The oxygen-gradient diffusion at capillary tissue level is essential for the cellular survival, while the homeostasis of the oxygen in the arterial blood is mediated by reflexes sensitive to oxygen decrease and by release of several factors. Aging is correlated with a reduction of cells' oxygen supply concomitant to a parallel decrease in oxygen demand by tissues. Both chronic hypoxia or hyperoxia are considered as stresses. Indeed, in both conditions, free radical species, which damage structural and functional components of the membrane, are generated. ROS (reactive oxygen species) are physiological products of aerobic life and their accumulation affects aging. Because hypoxia per se modulates mitochondria activity, influencing oxygen consumption, hypoxia and aging could share some link. Moreover, the observation that in hypoxia or hyperoxia there is an accumulation of lipofucsine as a general reaction to stress is consistent with the accumulation of such components during aging. Correlation between hypoxia-hyperoxia and life-span remains open until we solve the question of how and why do cells sense oxygen. In other words, to better understand aging we need to know what O(2) species are being sensed by cells. In conclusion, hypoxia and hyperoxia represent an experimental model adequate for studying aging processes. PMID- 20021404 TI - Dominance variation in the correlation between longevity and heat-stress resistance in Drosophila melanogaster. AB - The possible genetic correlation between longevity and knockdown resistance to high temperature (KRHT) was tested in reciprocal backcrosses between nearly homozygous lines of D. melanogaster. These baselines differ dramatically for KRHT. Because nearly-homozygous lines were used, reciprocal backcrosses were informative to test for dominant versus additive genetic associations between the traits. The line of high KRHT was longer lived than the low KRHT line. The correlation between longevity and KRHT was dependent on the backcross (BC). When a BC was set up with males from the low KRHT line, the most KRHT resistant individuals were in turn the longest-lived flies. However, this was not found in the reciprocal backcross, indicating dominance. Alleles that confer increased KRHT are dominant in the correlation between longevity and KRHT. The between-line variation in the traits studied is consistent with the hypothesis that stress resistant genotypes correspond to long-lived individuals. PMID- 20021405 TI - Tissular senescence and modifications of oral ecosystem in the elderly: risk factors for mucosal pathologies. AB - The aim of this present review is to describe the pathogenesis and mechanisms behind mucosal pathologies in the elderly including a description of the risk factors for these pathologies. The oral cavity - and particularly oral mucosae - is exposed to many stresses as well as physical, chemical, thermic and pathogenic agents. In the elderly, mucosae are less resistant to the insults, and this increases the occurrence of diseases. Several factors contribute to the prevalence of mucosal pathologies with aging. There are two categories: intrinsic factors linked to the senescence of the tissues and functions, and extrinsic factors related to the older people general health status. The intrinsic factors are: 1) mucosal senescence which induces fragility 2) immunosenescence which causes a decrease in the host response against micro-organisms and an increase in the autoimmune diseases and 3) senescence of salivary glands and reduction of the saliva protective function. Furthermore, there are extrinsic factors which contribute to change the oral ecosystem during aging, such as polypathologies and polymedications, malnutrition, degradation of oral hygiene, pathogen proliferation (mainly bacteria and Candida species) and old or ill-fitted removable dentures. In the elderly several diseases occur on the oral mucosae: inflammation, bacterial infections or candidiasis, ulcerations, autoimmune dermatosis, tumoral processes. This review describes some common oral mucosal pathologies in the older people, which illustrate the impact of different risk factors described in the first part. PMID- 20021406 TI - Aging and contact dermatitis: a review. AB - As the population continues to age, understanding the differences between aged and young skin becomes more important to the clinician. Contact dermatitis is just one of the clinical entities that presents differently in the geriatric population. Reactions can be delayed in onset, and prolonged in duration. These altered presentations are the direct result of the pathophysiologic changes that occur in aging skin. It is especially important to recognize contact dermatitis in this age group, and to treat in a timely fashion. PMID- 20021407 TI - Homology modeling of adenosine A2A receptor and molecular docking for exploration of appropriate potent antagonists for treatment of Parkinson's disease. AB - Parkinson's disease (PD) is a neurodegenerative disorder of central nervous system (CNS) that impaired the patient motor skills, speech and other functions. Adenosine A2A receptors have a unique cellular distribution in the neuron, which is used as a potential target for PD. Homology modeling was used to construct the 3-D structure of A2A using the known template (PDB: 2VT4), and the stereochemical quality was validated. Several effective antagonist drugs were selected and active amino acid residues in A2A were targeted on the basis of robust binding affinity between protein-drug interactions in molecular docking. Six antagonists, Bromocriptine, Cabergoline, Etilevodopa, Lysuride, Melevodopa and Pramipexole, were found more potent for binding and the active amino acids residues were identified (http://www.rcsb.org/pdb/) in A2A receptor. It could be used as the basis for rationale designing of novel antagonist drugs against Parkinson's disease. PMID- 20021408 TI - Age-related changes in posture control are differentially affected by postural and cognitive task complexity. AB - The simple postural task of quiet standing, which requires minimal attentional resources, is generally paired with cognitive activity. Competition for attentional resources is a consequence of simultaneously performing balance tasks and cognitive tasks, and impairment of attentional resource allocation with aging leads to increased risks of fall. We investigated age-related changes in posture control during dual task performance, using a paradigm that crossed a static (quiet standing) and a dynamic (keeping balance on a translational force plate) postural task and cognitive tasks of low demand (mental arithmetic) and high demand (spatial memory). Postural performance was analyzed through center-of pressure displacements using both statistical (body sway area/sway velocity) and nonlinear (wavelet transform) methods in three age groups (younger, middle-aged, and older healthy participants). Results showed that 1) the nonlinear analysis method was more sensitive than the traditional approach in distinguishing performance between age groups, a result that explains discrepancies in the dual task literature; 2) dual-tasking costs were dependent on both postural task difficulty and cognitive task complexity, corroborating previous investigations; 3) younger adults improved their postural performance during dual-tasking, but older adults lowered their performance; 4) balance recovery strategies in the dynamic postural task appeared to differ in younger versus older adults. Together, our findings on dual-tasking can be interpreted within the conceptual frame of task prioritization: shifting attention away from postural task automates posture control in the younger adults, whereas prioritization of postural task and selection of compensatory strategy are the main characteristics of posture control in the older population. PMID- 20021409 TI - Bedside balance testing in elderly people. AB - Balance impairment in older people is a problem for both patients and physicians. Patients suffer from dizziness and fear of falling, and as many of them also have mobility problems the risk of falling is higher than in younger patients with balance impairment. Balance testing in young patients is often a time-consuming task requiring expensive devices such as videonystagmography, posturography or evoked potentials. The task is more difficult in elderly patients, who have mobility problems, hearing loss, chronic pathology, medication, etc., or who cannot afford expensive testing. In this paper, we briefly review the physiopathology of the balance system considered as a data-processing system. Based on these concepts, we review the main tests that can be easily performed at bedside or in the patient's home using no devices at all. These tests are basic oculomotor testing (like spontaneous nystagmus, head impulse test, dynamic visual acuity, head-shaking nystagmus), provocation tests (like Dix-Hallpike manoeuvre and variants for positional vertigo). Finally, stance tests with a special focus on those which can identify altered postural control patterns and high risk of falling (like the modified Clinical Test for Sensory Interaction in Balance and the Timed Up and Go test). We comment their utility, how they should be performed and what results can be obtained. PMID- 20021410 TI - The pathogenesis of susceptibility to acute kidney injury in the elderly. AB - Elderly patients, especially those with chronic kidney disease (CKD), are at high risk for the development of acute kidney injury (AKI). AKI can lead to longer hospital stays, higher mortality rates and an increased risk for the development of end-stage renal disease. Clinical variables such as combordid states, impaired renal function, polypharmacy and an increase in high-risk procedures account for some of the increased risk for AKI in elderly patients. However, specific structural, functional, hemodynamic and cellular changes that occur with aging predispose the kidney to injury in stressful states. Understanding the interactions of these intra-renal changes with aging offers the opportunity to design specific strategies that can lower the risk for the development of AKI and its complications. PMID- 20021411 TI - Aging, mitochondria and male reproductive function. AB - The rise in life expectancy over the last century, together with higher maternal and paternal ages and have highlighted the issue of reduced fertility with advancing age. Aging of the male reproductive system is incited by multi factorial changes at molecular, cellular and regulatory levels, and individual characteristics are highly variable, although strongly influenced by lifestyle and environmental factors. Damage accumulated with age leads to progressive deregulation of the hypothalamic-pituitary-gonadal axis and of local auto/paracrine interactions, thereby inducing changes in target organs such as the testis, penis and prostate. Elderly human males produce less testosterone, have fewer motile sperm and a higher incidence of erectile dysfunction and prostate disorders, all of which contribute to lower fertility. Cellular aging can manifest itself at several levels. Aging cells progressively accumulate "waste" products, resulting in a decreased functionally. Changes to mitochondria are among the most remarkable features observed in aging cells and several theories place mitochondria at the hub of cellular events related to aging, namely in terms of the accumulation of oxidative damage to cells and tissues, a process in which these organelles may play a prominent role, although alternative theories have also emerged. Furthermore, mitochondrial energy metabolism is also crucial for male reproductive function and mitochondria may therefore constitute a common link between aging and fertility loss. PMID- 20021412 TI - Age-related changes in P-glycoprotein expression in senescence-accelerated mouse. AB - P-glycoprotein, the gene product of ATP-binding cassette, sub-family B (Abcb1), is a representative efflux transporter of cerebral vessels. It was recently reported that the expressions of P-glycoprotein and Abcb1 gene were increased in hippocampal vessels with blood-brain barrier (BBB) damage in stroke-prone hypertensive rats. SAMP8, senescence-accelerated mice with age-related deficits in memory and learning, are known to show age-related damage of BBB. Accordingly, in this study, we examined the P-glycoprotein expression and the gene expression (Abcb1a/b) by real-time quantitative reverse transcriptase-polymerase chain reaction (RT-PCR), Western blotting, and immunohistochemical techniques. SAMR1, which has a spontaneous retroviral insertional mutation in Abcb1a gene, was used to assess the effects of Abcb1a gene mutation. The brain samples of SAMR1 showed decreased expressions of P-glycoprotein and Abcb1a genes and increased expression of Abcb1b gene, compared with those of SAMP8 mice. The P-glycoprotein expression increased with aging in the brain samples of SAMP8, but not in those of SAMR1. The gene expressions of Abcb1a and Abcb1b increased with aging in the brain samples of SAMP8. Immunosignals of P-glycoprotein were seen in vessel walls, mainly in the cytoplasm of CD34-positive endothelial cells and partially in astrocytes, in all mice. These findings indicate that the expressions of Abcb1a and Abcb1b genes and their gene products, P-glycoprotein, were increased with aging in SAMP8, suggesting age-related response to prevent toxic substance from accumulating in the brains of SAMP8. PMID- 20021413 TI - Mortality according to a prior assessment of biological age. AB - BACKGROUND: Measures of biological age have not been proven to predict mortality. This study examines whether measuring biological age improves the prediction of mortality. METHODS: Prospective study from 1981 to 2001 of 397 male London Civil Servants. Two indices of biological ageing were calculated. RESULTS: 60 men died and both indices of biological ageing were related to survival. In a model that mutually adjusted for both chronological and biological age, biological age using index one was statistically significant with a hazard ratio (HR) of 1.11 per year of age (95% confidence interval 1.01 - 1.21, P=0.03). The useful components of the measures of biological ageing were systolic blood pressure (HR 1.31 for 1SD), albumin, and, to a lesser degree, Erythrocyte Sedimentation Rate (ESR). Greying of the hair, skin inelasticity, arcus senilis, and baldness were not predictors of mortality as measured by our methods. Similarly serum cholesterol, creatinine, calcium and urate could be excluded. A modified index was developed including systolic pressure, ESR, urea, albumin, and bilirubin and had a sensitivity of 78% and specificity of 51% in predicting subjects who died. CONCLUSION: This study represents 'proof of principle' in demonstrating the utility and validity of measuring biological age. The modified index needs to be tested prospectively. PMID- 20021414 TI - Comparing age effects in normally and extremely highly educated and intellectually engaged 65 - 80 year-olds: potential protection from deficit through educational and intellectual activities across the lifespan. AB - Education and cognitive activity have been suggested to protect against cognitive decline in old age. However, little is known about the long-term effects of extremely high levels of education and intellectual activity across the lifespan. The present study investigated the extent to which these two variables may moderate the age-related differences in cognitive performance in old adults. Therefore, story recall, paired-associates learning, reading span and letter digit performance of 62 university professors (mean age = 72.47) were compared with those of a representative sample of 196 participants of the Zurich Longitudinal Study of Cognitive Aging (mean age = 73.04). The results demonstrate that the highly educated sample performed significantly better than the normally educated sample in the paired-associates learning and reading span test. Furthermore, age effects were found in the letter digit as well as in the paired associates learning test. While the normally educated sample demonstrated an age related decrease in the paired-associates learning test, the performance of the highly educated sample actually increased with increasing age. These findings suggest that extremely high levels of education and intellectual activity may postpone age-related deficits in paired-associates learning tasks, but not in speed of processing tasks. PMID- 20021415 TI - Pathology associated memory deficits in Swedish mutant genome-based amyloid precursor protein transgenic mice. AB - To gain insight into the relationship between pathological alterations and memory deficits observed in Alzheimer's disease (AD), a number of amyloid precursor protein (APP) transgenic animal models have been generated containing familial AD mutations. The most commonly utilized method involves a cDNA-based approach, utilizing heterologous promoters to drive expression of specific APP isoforms. As a result of the assumptions inherent in the design of each model, the different cDNA-based transgenic mouse models have revealed different relationships between the biochemical, pathological and behavioral alterations observed in these models. Here we provide further characterization of a genomic-based, amyloid precursor protein yeast artificial chromosome transgenic mouse model of AD, R1.40, that makes few assumptions regarding disease pathogenesis to study the relationship between brain pathology and altered behavior. Aged R1.40 transgenic and control mice were tested for learning and memory in the Morris water maze and for working memory in the Y maze. Results from the water maze demonstrated intact learning in the both control and R1.40 mice, but impairments in the long-term retention of this information in the transgenic mice, but not controls. Interestingly, however, long-term memory deficits did not correlate with the presence of Abeta deposits within the group of animals examined. By contrast, age related working memory impairments were also observed in the Y maze in the R1.40 mice, and these deficits correlated with the presence of Abeta deposits. Our results demonstrate unique behavioral alterations in the R1.40 mouse model of AD that are likely both dependent and independent of Abeta deposition. PMID- 20021416 TI - Relationship of dietary habits and obesity to oxidative stress in Palauan people: compared with Japanese and Mongolian people. AB - The Republic of Palau belongs to Micronesia, and obese people and lifestyle related diseases are prevalent there. We investigated the relationship of dietary habits and obesity to oxidative stress in Palauan people, as compared with those of Japanese and Mongolian people. A total of 126 healthy Palauan subjects were enrolled. Oxidative stress was evaluated by serum level of reactive oxygen metabolites (ROM). Antioxidant capacity was evaluated by serum level of biological antioxidant potential (BAP). In Palauan subjects, BMI> or =30 was observed in 45.0% of males and 59.1% of females (Japanese: males 1.3%, females 0.8%, Mongolian: males 6.3%, females 14.7%). Palauan subjects consumed 2553 kcal per day (Japanese 2121 kcal, Mongolians 2534 kcal). The ratios of carbohydrate energy to total energy were 59.8 % (Japanese 54.7 %, Mongolians 50.2%). The ratios of fat energy to total energy were 22.9% (Japanese 26.7%, Mongolians 34.5%). ROM levels in Palauan subjects showed higher than those in Japanese subjects, while BAP levels of Palauan subjects did not decrease compared to those of Japanese. ROM levels correlated with body fat ratio, and showed a reverse correlation with handgrip strength. Handgrip strength decreased in the subjects of more than forty years of age. These findings suggest that the obesity in Palauan people may have a connection with high intake of calories through carbohydrate eating rather than through fat eating. Their high oxidative stress may be induced by obesity, and contribute to an early decline of handgrip strength, ultimately in early aging. PMID- 20021417 TI - Translational pharmacology in aging men with benign prostatic hyperplasia: molecular and clinical approaches to alpha1-adrenoceptors. AB - Benign prostatic hyperplasia (BPH) occurs in over 50% of men over 60 years of age, most of whom have lower urinary tract symptoms (LUTS). The incidence of BPH appears to be increasing due to the increased longevity of men. LUTS associated with BPH (LUTS/BPH) greatly affects the patient's quality of life. Physicians should be aware of other issues associated with advancing age, such as cardiovascular diseases, sexual dysfunction and cataract, that may complicate the treatment of aging men with LUTS/BPH; therefore, management of LUTS/BPH requires careful selection of the most appropriate treatment for each patient. Alpha1 adrenoceptor (alpha(1)-AR) stimulation plays an important role in the regulation of prostate smooth muscle contraction. Recent remarkable advances in molecular biology offer the possibility of new findings about the role of alpha(1)-AR subtypes and new therapeutic options for BPH; however, even though alpha(1)-AR antagonists are used as the first-line medical treatment for patients affected by LUTS/BPH, responses to alpha(1)-AR antagonists differ among patients. The risks of unexpected adverse events, acute urinary retention and the need for invasive therapy also differ among patients. Translational pharmacology is the collaboration between researchers and clinicians to discover more effective medical therapies and to identify new drugs for various diseases. This review summarizes the recent molecular and physiological findings of alpha(1)-AR subtypes and discusses potential new strategies for BPH medical treatment. In the future, the promise of genetic-based prescriptions and therapeutic plans as a useful strategy to improve clinical outcomes of BPH medical therapy may become credible and warrants further investigation. PMID- 20021418 TI - Short- and long-term survival of nonsurgical intensive care patients and its relation to diagnosis, severity of disease, age and comorbidities. AB - OBJECTIVE: To identify variables associated with mortality in the ICU and 1 year following discharge. DESIGN: Prospective observational cohort study. SETTING: ICU of a tertiary care center and university hospital. PATIENTS: A total of 3,119 medical and neurological intensive care patients. MEASUREMENTS AND MAIN RESULTS: Pre-admission health status was quantified by the sum of risk factors and chronic diseases. Severity of the acute disease was estimated by counting the number of organ dysfunctions and the Acute Physiology Score. Concerning the primarily affected organ system, ICU mortality was highest in hematological diseases (63%) and 1-year mortality was 82%. Lowest death rates were observed with metabolic (ICU 4%, 1-yr 18%) and psychiatric diagnoses (ICU 5%, 1-yr 13%). Greater severity of illness with the need for mechanical life support was associated with decreased 1-year survival. In the respiratory and in renal diseases, the artificial support of the primarily affected organ system incurred an ICU mortality equaling the average (23%) or below (14%) that of the whole ICU population. Pre-admission health status increased the probability of developing multiple organ failure and worsened outcome 1 year after discharge in non cardiovascular patients. Age showed a weak correlation with chronic diseases and severity of the acute illness and was related to long-term, but not short-term survival. CONCLUSIONS: The most important risk factors associated with short- and long-term mortality in non-surgical intensive care patients are disease severity and the primarily affected organ system that necessitates admission. The artificial support of this organ system can improve only short-term outcome. PMID- 20021419 TI - Organ- and cell-type specific delivery of kinase inhibitors: a novel approach in the development of targeted drugs. AB - During the past years, we have explored the cellular delivery of kinase inhibitors. Kinase inhibitors have selectivity for specific kinases but they lack cellular selectivity. This is exemplified by recent reports on cardiotoxicity of kinase inhibitors used in cancer treatment. We postulate that targeted cellular delivery of kinase inhibitors can improve their safety/toxicity profiles, as will be exemplified by recent published studies. Cell specific delivery of therapeutics is a quickly growing area of investigation. This innovative strategy employs carrier molecules that bind to receptors exposed on the surface of cell types involved in disease processes. Binding and receptor mediated internalization of the carrier facilitates local accumulation of the product in target cells. Upon systemic administration, this may create local drug depots in specific organs, while other tissues are avoided, thus favoring enhanced localized drug efficacy and reduced side-effects. Synthesis of targeted kinase inhibitor-carrier conjugates was achieved using a new approach, in which kinase inhibitors were bound to a platinum(II) atom, the so-called Universal Linkage System (ULS). We review this novel linkage chemistry and demonstrate the applicability of ULS for drug targeting approaches aiming at angiogenic endothelial cells, hepatic stellate cells, and kidney tubular cells. We will review important issues like drug release mechanism, safety of the linker, and pharmacokinetics of the products in animals. Finally, we review the pharmacological efficacy of the cellular targeted drug conjugates in experimental animal models, especially in renal and liver fibrosis models. PMID- 20021420 TI - Gene therapy: the first approved gene-based medicines, molecular mechanisms and clinical indications. AB - As gene therapy has matured from clinical trials to the first commercial products, understanding of the mechanisms of gene delivery has increased tremendously. This has also been reflected in viral vector development, creating a number of new approaches to tackle issues in transduction efficiency, biodistribution and viral safety. This review will highlight the most important issues and advancements in vector development, administration, surface modification, integration to host genome and safety. The gene therapy products currently available or near market approval, based on p53 expression (Gendicine and Advexin), conditionally replicative adenoviruses (Oncorine) and thymidine kinase + ganciclovir therapy (Cerepro), are introduced with emphasis on the molecular mechanisms of action. PMID- 20021421 TI - Inhibitors of the Ubiquitin-Proteasome System and the cell death machinery: How many pathways are activated? AB - Over the past decade, the promising results of UPSIs (UPS inhibitors) in eliciting apoptosis in various cancer cells, and the approval of the first UPSI (Bortezomib/Velcade/PS-341) for the treatment of multiple myeloma have raised interest in assessing the death program activated upon proteasomal blockage. Several reports indicate that UPSIs stimulate apoptosis in malignant cells by operating at multiple levels, possibly by inducing different types of cellular stress. Normally cellular stress signals converge on the core elements of the apoptotic machinery to trigger the cellular demise. In addition to eliciting multiple stresses, UPSIs can directly operate on the core elements of the apoptotic machinery to control their abundance. Alterations in the relative levels of anti and pro-apoptotic factors can render cancer cells more prone to die in response to other anti-cancer treatments. Aim of the present review is to discuss those core elements of the apoptotic machinery that are under the control of the UPS. PMID- 20021422 TI - Apoptosis-regulatory factors as potential drug targets in the epithelium of normal and inflamed airways. AB - Airway epithelium (AE) lines the conducting airways of the respiratory system and functions to maintain airway integrity by providing both a physical barrier to inhaled noxious agents and a mechanism for their clearance via the mucociliary escalator. Normal AE cells are relatively refractory to a number of apoptotic stimuli and survival mechanisms are in place to maintain the integrity of the epithelial barrier that is exposed to agents such as reactive oxygen species (ROS) and death receptor ligands secreted by immune cells during inflammation. When damage to AE does occur, there is increased AE apoptosis, such as in the airway damage that occurs in the chronically inflamed airways in diseases like asthma where rates of AE apoptosis can be increased many-fold. The usual treatment for persistent asthma in humans involves a combination of bronchodilator and inhaled corticosteroid; there is however a need to develop strategies to better control other aspects of the disease, including minimizing the ongoing damage to AE and consequent airway remodeling. Targeting of the major apoptosis-regulatory factors in AE may be one such strategy. Here we review what is known about apoptosis and its regulatory factors in normal AE and abnormalities in these factors in the inflamed airways of mice and humans. PMID- 20021423 TI - Pharmacology and therapeutic applications of enediyne antitumor antibiotics. AB - The natural compounds that interfere with cellular DNA such as enediyne antitumor antibiotics might be important chemotherapeutic agents for the treatment of cancer. In this article, the pharmacology and anticancer activity of the enediyne antitumor agents that are approved for clinical use and undergoing pre-clinical or clinical evaluation are reviewed. Most enediyne compounds have shown potent activity against the proliferation of various cancer cells, including cells that display resistance to other chemotherapeutic drugs. Enediyne derivatives, such as an immunoconjugate composed of an enediyne compound and monoclonal antibody, reveal stronger activity and selectivity for human cancer cells. The mechanism underlying the anticancer activity of these enediyne antitumor agents may mainly lie in their generation of DNA double-strand breaks. Increasing evidence shows that the enediyne-induced DNA double-strand breaks can engage the activation of DNA damage response proteins, arresting cell cycle progression and eventually leading to apoptotic cell death. Continued investigation of the mechanisms of action and development of new enediyne derivatives and conjugates may provide more effective therapeutics for cancer treatments. PMID- 20021424 TI - Pharmacologic intervention in axonal excitability: in vivo assessment of nodal persistent sodium currents in human neuropathies. AB - Axonal excitability testing can provide new insights into the ionic mechanisms underlying the pathophysiology of hyperexcitability of motor and sensory axons in human neuropathies. Threshold tracking was developed in the 1990's to non invasively measure a number of axonal excitability indices that depend on sodium and potassium channel function, and this makes it possible to monitor the effects of pharmacologic intervention with ion channel modulators. This paper reviews recent advances in ionic-pathophysiological studies in humans. (1) Neuropathic pain or muscle cramp/fasciculation is partly caused by hyperexcitability of the injured axons. The enhanced excitability can result from altered ion channel function; such as an increase in persistent sodium currents. Persistent sodium currents can be reliably estimated using threshold tracking. In peripheral neuropathy, persistent sodium currents usually increase possibly due to over expression of sodium channels associated with axonal regeneration, and could be responsible for ectopic firings. Administration of sodium channel blockers such as mexiletine, results in marked alleviation of muscle cramping in parallel with a decrease in nodal persistent sodium currents. (2) In diabetic neuropathy, the activation of the polyol pathway mediated by an enzyme, aldose reductase, leads to reduced Na(+)/K(+) pump activity, and intra-axonal sodium accumulation; sodium currents are reduced presumably due to decreased trans-axonal sodium gradient. Aldose reductase inhibitiors improve nodal sodium currents, as well as nerve conduction, and this can be objectively assessed by threshold tracking. Studies of ion-channel pathophysiology in human subjects have recently begun. Investigating ionic mechanisms by monitoring the corresponding ionic currents. is of clinical relevance, because once a specific ionic conductance is identified, pharmacologic blocking or modulation could provide a new therapeutic option. PMID- 20021425 TI - Immunomodulatory properties of antibiotics. AB - There is growing evidence that certain antibiotics exert their beneficial effects not only by killing or inhibiting the growth of bacterial pathogens but also indirectly by immunomodulation. This review aims at giving an overview of the immunomodulatory properties of antibiotics in different diseases: The antiinflammatory properties of macrolides in chronic inflammatory pulmonary disorders were recognized more than 15 years ago and have been well documented in the last decade. Recent data suggest that several antibiotics such as tetracyclines and cephalosporins may have a beneficial immunomodulatory or neuroprotective effect on neuroimmunological and neurodegenerative diseases including multiple sclerosis and amyotrophic lateral sclerosis. Moreover, the non bacteriolytic but bactericidal antibiotics rifampicin, clindamycin and aminoglycosides kill bacteria without releasing high quantities of proinflammtory cell wall components. The use of bactericidal, non-bacteriolytic protein synthesis inhibitors reduces mortality and long-term sequelae in experimental bacterial sepsis, plague and meningitis. Clinically, macrolides have been well established as an adjunctive treatment to beta-lactam antibiotics in pulmonary diseases. For other indications, appropriate clinical trials are necessary before using the immunomodulatory properties of antibiotics in clinical practice. PMID- 20021426 TI - Targeting cancer cells by an oxidant-based therapy. AB - Despite the progress achieved in chemo- and radiotherapy, cancer is still a leading life-threatening pathology. In that sense, there is a need for novel therapeutic strategies based on our current knowledge of cancer biology. Among the phenotypical features of cancer cells, two of them are of particular interest: their nearly universal glycolytic phenotype and their sensitivity towards an oxidative stress, both resulting from the combination of high anabolic needs and hypoxic growth conditions. By using menadione (vitamin K3) and ascorbate (vitamin C), we took advantage of these features to develop an original approach that consists in the exposure of cancer cells to an oxidant insult. When used in combination, these compounds exhibit a synergistic action and are devoid of major toxicity in vivo. Thus, this review is dedicated to the analysis of the molecular pathways by which this promising combination exerts its antitumoural effect. PMID- 20021427 TI - Modulation of transmitter release via presynaptic ligand-gated ion channels. AB - Neurons communicate through the exocytotic release of transmitters from presynaptic axon terminals and the ensuing activation of postsynaptic receptors. Instantaneous responses of postsynaptic cells to released neurotransmitters are mediated by ligand-gated ion channels, whereas G protein-coupled receptors mediate rather delayed effects. Moreover, the actions of ionotropic receptors are transient (milliseconds to seconds) and those of G protein-coupled receptors are more long lasting (seconds to minutes). Accordingly, neuronal signalling via ligand-gated ion channels is termed neurotransmission, whereas signalling via G protein-coupled receptors is termed neuromodulation. Exocytotic transmitter release is modulated by a variety of mechanisms such as previous activity at the synapse and the presence of extracellular neurotransmitters. Like the postsynaptic responses, presynaptic modulation is not only mediated by slowly acting G protein-coupled receptors, but also by fast acting ligand-gated ion channels. Accordingly, members of all known families of ligand-gated ion channels (cys-loop receptors, such as GABA(A), glycine, nicotinic acetylcholine, and 5 HT(3) receptors, ionotropic glutamate receptors, P2X receptors, and vanilloid receptors) are known to control transmitter release. All these ligand-gated ion channels display heterogeneous structures and functions. Therefore, activation of such presynaptic receptors can control transmitter release in different ways and through a multitude of mechanisms. This review provides a summary of the functions of the different presynaptic ligand-gated ion channels and presents prototypic examples for the physiological and pharmacological relevance of these presynaptic receptors. PMID- 20021428 TI - Emerging therapeutic strategies for hepatitis C virus infection. AB - The universal prevalence of hepatitis C virus (HCV) infection, which causes chronic hepatitis, cirrhosis, liver failure, and hepatocellular carcinoma, has become a significant health problem worldwide. Interferon-based therapies, the current standard, IFN-based therapies have limited efficacy and undesirable adverse effects. In addition, neither vaccination against HCV nor specific antiviral reagents for HCV are yet available. Thus, a major medical need still exists for novel and more efficacious anti-HCV reagents showing broad-spectrum clinical efficacy with enhanced tolerability. With the progress in our current understanding of the function and regulation of HCV gene products, the three dimensional structures of virally encoded enzymes and the recent establishment of the HCV-replicon system, several pharmacological targets are being studied for HCV therapy, including cellular receptors mediating HCV entry, factors facilitating HCV replication and assembly, and intracellular pathways. Recently developed mouse models will be very helpful in evaluating the in vivo efficacy of novel antiviral reagents. Currently many novel antiviral drugs are under evaluation in clinical trials. This review will comprehensively discuss the current treatment options and various novel antiviral reagents available. Ongoing clinical studies of promising lead drugs are also reviewed. PMID- 20021429 TI - Diamidine activity against trypanosomes: the state of the art. AB - Aromatic diamidines and related compounds are DNA minor groove binders that have been screened against a variety of pathogenic microorganisms such as bacteria, fungi and protozoa and show promising results. Parasitic infections are widespread in developing countries and are major contributors to human mortality and morbidity, causing considerable economic hardship. Trypanosomes are unicellular protozoan organisms that cause serious public health problems in developing countries: African trypanosomiasis (sleeping sickness) in Africa, and Chagas' disease, in Latin America. Sleeping sickness, caused by sub-species of Trypanosome brucei (T. brucei gambiense and T. brucei rhodesiense), is a fatal disease if left untreated, with about 60 million people currently at risk. Trypanosoma cruzi is the etiological agent of Chagas' disease, an important parasitic illness that affects nearly 17 million individuals in endemic areas. The fact that the available clinical drugs are expensive, toxic, require long treatment periods, frequently exhibit reduced activity towards certain parasite strains and evolutive stages, and are beginning to show development of resistance, demonstrates the urgent need for the development of new drugs for both pathologies. For some time much attention has been focused on the effect of diamidines (and related compounds) on African trypanosomes. However more recent studies have pointed to their potential activity against T.cruzi. In this review the current therapeutic state of the art of aromatic diamidines and related compounds used against T.brucei and T.cruzi is reviewed with a focus on their potential use as antiparasitic drugs for the treatment of both these important neglected diseases. PMID- 20021430 TI - DNA polymerases and oxidative damage: friends or foes? AB - DNA is modified by many mutagens, including reactive oxygen species (ROS). When ROS react with DNA, various kinds of modified base and/or sugar moieties are produced. One of the most important oxidative DNA lesions is 7,8-dihydro-8 oxoguanine (8-oxo-G). Contrary to normal deoxyguanosine, 8-oxo-G favors a syn conformation, enabling it to form a Hoogsteen base pair with adenine which resembles a normal Watson-Crick base pair in shape and geometry. As a consequence, most human DNA polymerases (pols) studied so far show significant error-prone bypass of 8-oxo-G. The 1,2-dihydro-2-oxoadenine (2-OH-A) is another common DNA lesion produced by ROS. 2-OH-A possesses significant mutagenic potential in living cells. When challenged with a 2-OH-A lesion on the template, DNA pols often misinsert G and C nucleotides, with various efficiencies depending upon the sequence context. We have recently shown that human DNA pol lambda is extremely efficient in performing error-free bypass of both 8-oxo-G and 2-OH-A lesions, and that its efficiency is positively modulated by the auxiliary factors proliferating cell nuclear antigen and replication protein A. In this review we will summarize the most recent advancements in the field of oxidative DNA damage tolerance with special emphasis on the pro- and anti-mutagenic roles of DNA pols and auxiliary proteins. PMID- 20021431 TI - Host genetic factors and treatment of hepatitis C. AB - Infection with the hepatitis C virus (HCV) is a major health problem worldwide due to the associated risk of developing liver cirrhosis and its sequelae. Approximately 200 million persons are chronically infected worldwide. Furthermore, about one third of HIV-infected individuals in Europe and the US are co-infected with HCV. Currently pegylated interferon-alpha in combination with ribavirin represents the backbone of HCV-specific therapy. However, with interferon-based combination therapy sustained virologic response (SVR) is achieved in only about 50% of HCV-infected patients in clinical studies and may be even lower in clinical practice. HCV genotype and viral load are major determinants of treatment response in HCV infection. However, emerging data suggest host genetic factors also influence response to treatment. These data might hold the keys to better understand and predict outcome of HCV-specific therapy and might help to develop novel anti-HCV strategies. Here, we review the role of genetic aspects including the role of cytokines, chemokines/ chemokine receptors, and MHC alleles with respect to HCV therapy that have been elucidated so far and offer suggestions for how to use these observations as platforms for future research to further understand differential response to antiviral therapy in HCV-infected patients. PMID- 20021432 TI - Neuropharmacology of the olfactory bulb. AB - The olfactory bulb is located at the start of a hierarchical chain of sensory processing mechanisms. The relative ease of its isolation allows the possibility that models of these mechanisms might be integrated to develop a detailed understanding of function. In this sensory processing chain odour molecules evoke signal transduction in the olfactory receptor neurons. These signals represent the diverse range of molecular binding affinities of the olfactory receptor proteins. The first level of processing of this sensory input is performed by the neurons of the olfactory bulb. The olfactory system needs to filter the vast amount of sensory input it receives to be able to select the subset of biological significance. The importance of the olfactory bulb in this filtering process is suggested by its wide range of modulatory mechanisms. These mechanisms include an array of centrifugal inputs from other regions of the brain as well as numerous intrinsic feedback circuits. Given the complexity of the olfactory bulb and the range of its modulatory activity, the process of isolation of its components produces some difficulties of interpretation. This is mainly due to the removal of some of the effects of interaction and the change in balance that results. We present a summary of the current understanding of the interacting modulatory elements that are found in the olfactory bulb and a detailed account of the properties of mitral/tufted cells, the projection neurons of the olfactory bulb. This is followed by a discussion of the intrinsic and extrinsic modulatory systems acting on these cells. A consideration of the integration of the effects of these modulatory systems allows an understanding of how the output of the mitral/tufted cells is controlled. While significant progress has been made in the elucidation of the individual components as a result of advances in techniques over the last decade we suggest that there is a need for computational studies as a further aid to the understanding and interpretation of the weight of individual modulatory components in this dynamic interacting system. PMID- 20021433 TI - Hepatitis B vaccination in HIV-infected subjects. AB - Subjects at risk of infection with human immunodeficiency virus (HIV) are also at high risk of acute and chronic hepatitis B virus (HBV) infection. HIV is associated with higher HBV viraemia and with the risk of HBV reactivation, chronic active HBV infection, cirrhosis and death. Therefore, hepatitis B vaccination is recommended for all HIV-infected subjects lacking prior immunity. However, the immune response to hepatitis B vaccine is frequently suboptimal in this population. High CD4+ cell counts and low HIV viraemia are well known factors associated with a better rate of response. Moreover, higher hepatitis B vaccine doses and/or prolongation of the vaccination schedule, as implemented for patients with immune deficiencies other than HIV, may be considered. New vaccination cycles should be considered if post-vaccination titers of antibodies to hepatitis B surface antigen are < 10 mIU/mL (< 10 UI/L). The immunization of all young and middle-aged adults appears to be the most useful strategy to protect all patient-populations at high risk of sexually transmitted diseases. PMID- 20021434 TI - Biologic therapies against inflammatory bowel disease: a dysregulated immune system and the cross talk with gastrointestinal mucosa hold the key. AB - Inflammatory bowel disease (IBD) is a GI tract disorder that manifests as either Ulcerative colitis (UC) or Crohn's disease (CD). The precise etiology of IBD is still not completely elucidated but research into the immunopathogenesis of IBD suggests that dysfunctions of the intestinal immune system and cross-reactivity against host epithelial cells hold the key. In both UC and CD, polarized immune activity towards Th1 (marked by upregulation of TNF-alpha, IL-1beta, IFN-gamma, IL-6) and Th17 (marked by IL-17 secretion) response is reported, while UC appears to exhibit an added contribution of Th2 responses (characterized by secretion of IL-4, IL-5, and IL-13). Additionally, other molecules involved in leukocyte trafficking (adhesion molecules), chemokines (IL-8) and tissue repair molecules (PGE(2) and its receptors) are also crucial. Emergence of these new paradigms in the pathogenesis of IBD led to a recent trend of novel biological therapies that specifically inhibit molecules involved in the inflammatory cascade. In this review, we critically discuss recent advances in the pathogenesis of IBD, drug therapies (conventional versus biologic), drug efficacy and pharmacokinetics (murine versus human versus chimeric) and their adverse effects. We also discuss emerging novel biological therapies targeting pro-inflammatory cytokines including TNF-alpha and IFN-gamma, cytokine receptors and those targeting adhesion molecules-anti-integrin and anti-ICAM antibodies. Other potential approaches using anti-inflammatory cytokines (IL-10), anti-sense oligonucleotide and probiotics are also discussed. Finally, we summarized few imperative targets whose more detailed exploration can help to pave the way for an efficacious IBD therapy. PMID- 20021435 TI - Trends in the exploration of anticancer targets and strategies in enhancing the efficacy of drug targeting. AB - A number of therapeutic targets have been explored for developing anticancer drugs. Continuous efforts have been directed at the discovery of new targets as well as the improvement of therapeutic efficacy of agents directed at explored targets. There are 84 and 488 targets of marketed and investigational drugs for the treatment of cancer or cancer related illness. Analysis of these targets, particularly those of drugs in clinical trials and US patents, provides useful information and perspectives about the trends, strategies and progresses in targeting key cancer-related processes and in overcoming the difficulties in developing efficacious drugs against these targets. The efficacy of anticancer drugs directed at these targets is frequently compromised by counteractive molecular interactions and network crosstalk, negative and adverse secondary effects of drugs, and undesired ADMET profiles. Multi-component therapies directed at multiple targets and improved drug targeting methods are being explored for alleviating these efficacy-reducing processes. Investigation of the modes of actions of these combinations and targeting methods offers clues to aid the development of more effective anticancer therapies. PMID- 20021436 TI - Polyphenols: biological activities, molecular targets, and the effect of methylation. AB - Polyphenolic compounds are widely distributed in the plant kingdom and the anticancer benefits obtained from their consumption have been studied extensively. However, polyphenols are subject to various biotransformation reactions within the human body including methylation. Likewise, naturally occurring polyphenols may contain O-methylations in place of the hydroxyls of the parent compounds. While some studies suggest that methylations can increase the bioavailability of polyphenols, other studies indicate a decrease in the anticancer benefits of methylated polyphenols. This review will focus on the cellular activities of polyphenols, their potential molecular targets and their biological effects after enzymatic methylation. Furthermore, an assessment of the positive and negative aspects of polyphenol methylation on the anticancer activity will be discussed. Finally, the future of polyphenols in both cancer prevention and cancer intervention will be addressed. PMID- 20021437 TI - Pharmacological inhibition of the Bcl-2 family of apoptosis regulators as cancer therapy. AB - Conventional chemotherapy for cancer utilizes cytotoxic agents which elicit their therapeutic effect in part through the induction of apoptosis. In contrast, drugs which have been developed more recently and which are referred to as "targeted therapy" may exhibit less unwanted toxicity but in some cases these drugs are cytostatic. The recent development of drugs which target the apoptotic machinery offers a means to combine these two approaches. The intrinsic apoptotic pathway is controlled by the balance between anti-apoptotic proteins belonging to the Bcl 2 family and pro-apoptotic proteins bearing a single BH3 domain. Anti-apoptotic Bcl-2 family members are able to sequester the pro-apoptotic proteins by binding their BH3 domain. Compounds which inhibit this interaction are expected to promote apoptosis by preventing sequestration of the pro-apoptotic protein. Recently, a number of drugs have been developed which accomplish this, eg ABT 737, and some of these are progressing to clinical trials in oncology. These drugs may induce apoptosis on their own or synergize with existing chemotherapy. For example, ABT-737 is able to induce apoptosis when used as a single agent to treat leukemic and lung cancer cells and has also been shown to synergize with conventional chemotherapeutic agents in several cancer types. The spectrum of Bcl 2 family members expressed in a tumor cell, and the specificity of the inhibitor for these different anti-apoptotic proteins, helps determine whether Bcl-2 antagonists induce apoptosis when used as single agents. The ability of cytotoxic drugs to alter the expression of pro- and anti-apoptotic proteins is likely to help determine whether Bcl-2 antagonists synergize with cytotoxic therapy. Finally, as we begin to understand the pathways that regulate the expression of pro- and anti-apoptotic pathways, several new therapeutic strategies can be envisioned. PMID- 20021439 TI - Can increased food intake improve psychosis? A brief review and hypothesis. AB - Weight gain, diabetes, and changes in serum lipid profiles have been reported during treatment with typical and atypical antipsychotics. An association between diabetes and psychotic disorders was described long before the introduction of pharmacological agents for the treatment of schizophrenia. Several theories have been proposed to explain the baseline weight increase and metabolic disturbances in schizophrenia. Some studies suggest that increased food intake may improve psychotic symptoms in patients with schizophrenia but there have been conflicting results. Available clinical and basic research findings are discussed to evaluate the hypothesis that increased food intake may decrease sensitivity to dopamine signaling in the striatum. More research is needed to evaluate this potential link. However, basic animal research and evolutionary approaches can provide insights into metabolic disturbances associated with schizophrenia. PMID- 20021440 TI - Functional selectivity in cannabinoid signaling. AB - Cannabinoid (CB) agonists exhibit numerous potentially useful pharmacological properties, but unwanted side effects limit their use in clinical practice. Thus, novel strategies are needed to identify potential CB pharmaceuticals with fewer side effects. Activated CB receptors initiate multiple parallel intracellular signal transduction cascades. In the present paper we will review experimental data indicating that structurally different classes of CB agonists may exhibit selectivity toward individual subsets of intracellular signaling pathways. In support of this, recent findings indicate that chemically distinct classes of CB agonists frequently differ in their rank order of potency to produce analgesia versus other central nervous system effects in vivo. Structurally different agonists were also found to differ in their abilities to activate individual G protein types in vitro. Since it was suggested earlier that structurally distinct CB agonists may interact differently with the CB receptors, it has been hypothesized that different classes of cannabinoid agonists may stabilize unique active CB receptor conformations, leading to functional selectivity in CB receptor signaling. In order to obtain a direct proof for this hypothesis, we recently employed a highly sensitive biophysical method, plasmon-waveguide resonance (PWR) spectroscopy. PWR experiments have provided a direct proof that structurally different CB agonists produce qualitatively distinct changes in the shape and/or membrane orientation of the CB1 receptors, leading to functional selectivity in G protein activation. We expect that by identification of CB agonists that selectively activate preferred intracellular signaling pathways novel pharmacological lead structures can be identified for the design of improved CB analgesics with fewer side effects. PMID- 20021441 TI - Induced pluripotent stem cells, new tools for drug discovery and new hope for stem cell therapies. AB - Somatic cell nuclear transfer or therapeutic cloning has provided great hope for stem cell-based therapies. However, therapeutic cloning has been experiencing both ethical and technical difficulties. Recent breakthrough studies using a combination of four factors to reprogram human somatic cells into pluripotent stem cells without using embryos or eggs have led to an important revolution in stem cell research. Comparative analysis of human induced pluripotent stem cells and human embryonic stem cells using assays for morphology, cell surface marker expression, gene expression profiling, epigenetic status, and differentiation potential have revealed a remarkable degree of similarity between these two pluripotent stem cell types. This mini-review summarizes these ground-breaking studies. These advances in reprogramming will enable the creation of patient specific stem cell lines to study various disease mechanisms. The cellular models created will provide valuable tools for drug discovery. Furthermore, this reprogramming system provides great potential to design customized patient specific stem cell therapies with economic feasibility. PMID- 20021442 TI - Regulation of transcription factors by heterotrimeric G proteins. AB - Lessons from viral hijacks of cells and cancer biology suggest that the activation of G protein-coupled receptors (GPCRs) often results in the modulation of various transcription factors and cofactors. Since drugs acting on GPCRs represent a significant portion of therapeutic agents currently in use, it is important to understand the actions of GPCRs on gene expression. GPCRs and their associated heterotrimeric G proteins are known to regulate gene transcription through complex signaling networks. The G protein-mediated signaling cascades have been extensively studied and accumulating evidence indicates that the four subfamilies of G proteins may utilize both common and unique pathways for transcriptional regulation. This review aims to provide a contemporary account of our understanding on the regulation of transcription factors by GPCRs, with a special emphasis on specific regulations of transcription factors such as STAT3 and NF-kappaB by individual G protein subfamilies. Functional impacts of the signal integration between different pathways and the contributions by other GPCR interacting molecules will also be briefly discussed. PMID- 20021438 TI - TRPV1: on the road to pain relief. AB - Historically, drug research targeted to pain treatment has focused on trying to prevent the propagation of action potentials in the periphery from reaching the brain rather than pinpointing the molecular basis underlying the initial detection of the nociceptive stimulus: the receptor itself. This has now changed, given that many receptors of nociceptive stimuli have been identified and/or cloned. Transient Receptor Potential (TRP) channels have been implicated in several physiological processes such as mechanical, chemical and thermal stimuli detection. Ten years after the cloning of TRPV1, compelling data has been gathered on the role of this channel in inflammatory and neuropathic states. TRPV1 activation in nociceptive neurons, where it is normally expressed, triggers the release of neuropeptides and transmitters resulting in the generation of action potentials that will be sent to higher CNS areas where they will often be perceived as pain. Its activation also will evoke the peripheral release of pro inflammatory compounds that may sensitize other neurons to physical, thermal or chemical stimuli. For these reasons as well as because its continuous activation causes analgesia, TRPV1 has become a viable drug target for clinical use in the management of pain. This review will provide a general picture of the physiological and pathophysiological roles of the TRPV1 channel and of its structural, pharmacological and biophysical properties. Finally, it will provide the reader with an overall view of the status of the discovery of potential therapeutic agents for the management of chronic and neuropathic pain. PMID- 20021443 TI - Antiviral strategies: the present and beyond. AB - Historically, vaccine strategies have proven to be most effective at eradicating the targeted virus infections. With the advent of new or re-emerging altered viruses, some of which jump species to infect humans, the threat of viral pandemics exists. The protracted time to develop a vaccine during a pandemic necessitates using antiviral drugs in the intervening months prior to vaccine availability. Antiviral drugs that are pathogen specific, for example Amantidine, Tamiflu and Relenza, targeted against influenza viruses, are associated with the emergence of virus strains that are drug resistant. The use of ribavirin, a more broad spectrum antiviral, in combination therapies directed against influenza and hepatitis C virus, has proven effective, albeit to a modest extent. Attention is focused on the potential use of interferons (IFN)-alpha/beta as broad spectrum antivirals in acute infections, to invoke both direct antiviral effects against viruses and activation of specific immune effector cells. PMID- 20021444 TI - Immunological mechanisms of interleukin-2 (IL-2) treatment in HIV/AIDS disease. AB - HIV establishes a chronic infection that is marked by the progressive depletion of CD4+ T-cells, yet the mechanisms by which this depletion arises are a matter of controversy. Evidence is accumulating that T CD4+ depletion is not effected solely by virus-mediated killing and that mechanisms involving T-cell dynamics play a major role in the pathogenesis of HIV infection. Hence antiretroviral therapy, by controlling viral replication alone, invariably fails to achieve the broadest immune reconstitution. This issue has strengthened the rationale to widely explore new adjuvant immunotherapy. Most work has been performed on IL-2, given its potential to correct HIV-driven immune defects, possibly translating in a more effective immune competency. Important insights stem from the IL-2 mediated immune reconstitution pattern, with a rise in peripheral turnover and thymopoiesis, IL-7 synthesis and functional markers, resulting in the correction of the skewed T-cell immunophenotype and cytokine milieu. Combined, these findings suggest that IL-2 has a beneficial effect in correcting the severe disruption in T-cell homeostasis induced by HIV, through the interaction with T cells and cytokine microenvironment. However, whether or not these immunologic effects translate in an actual immunologic competency and therefore clinical benefit, still awaits demonstration from ongoing large, controlled clinical studies. PMID- 20021445 TI - Peroxisome proliferator-activated receptor (PPAR) beta/delta: a new potential therapeutic target for the treatment of metabolic syndrome. AB - Metabolic syndrome is defined as the clustering of multiple metabolic abnormalities, including abdominal obesity, dyslipidemia (high serum triglycerides and low serum HDL-cholesterol levels), glucose intolerance and hypertension. The pathophysiology underlying metabolic syndrome involves a complex interaction of crucial factors, but two of these, insulin resistance and obesity (especially visceral obesity), play a major role. The nuclear receptors Peroxisome Proliferator-Activated Receptors (PPAR)alpha and PPARgamma are therapeutic targets for hypertriglyceridemia and insulin resistance, respectively. Evidence is now emerging that the PPARbeta/delta; isotype is a potential pharmacological target for the treatment of disorders associated with metabolic syndrome. PPARbeta/delta; activation increases lipid catabolism in skeletal muscle, heart and adipose tissue and improves the serum lipid profile and insulin sensitivity in several animal models. In addition, PPARbeta/delta; ligands prevent weight gain and suppress macrophage-derived inflammation. These data are promising and indicate that PPARbeta/delta; ligands may become a therapeutic option for the treatment of metabolic syndrome. However, clinical trials in humans assessing the efficacy and safety of these drugs should confirm these promising perspectives in the treatment of the metabolic syndrome. PMID- 20021446 TI - Molecular basis of cardioprotection by erythropoietin. AB - Erythropoietin (EPO), a glycoprotein essential for red blood cell production acts on several non-erythropoietic tissues. The EPO receptor (EPOR) is expressed in a variety of cell types including neurons, endothelial cells, and cardiomyocytes. Recently, a number of reports have indicated that EPO preserves heart function in models of cardiac ischemia-reperfusion (I/R) injury. A diverse range of cellular/physiological processes is modulated by EPO and are thought to play a role in the preservation of heart function. In vivo, reductions in infarct size, apoptosis, oxidative stress, and inflammation have been reported. More recently, increases in angiogenesis and reductions in arrhythmias have been implicated in the cardioprotective effects of EPO. In vitro, EPO reduces apoptosis, oxidative stress, and inflammation. These cardioprotective effects appear to be mediated by a receptor interaction that is distinct from that responsible for EPO's erythropoietic effects. Downstream of receptor interactions, the activation of phosphatidylinositol-3 kinase (PI3-kinase) and Akt appear to mediate many of EPO's cardioprotective effects. However, there is emerging evidence for Akt independent mechanisms of cardioprotection including the inhibition of glycogen synthase kinase 3beta, as well as the activation of potassium channels, protein kinase C, and protein kinases such as ERK1/2. This review focuses on the effects of EPO in the heart and the molecular mechanisms by which EPO achieves its cardioprotective effects. PMID- 20021447 TI - p75NTR as a therapeutic target for neuropsychiatric diseases. AB - The p75 neurotrophin receptor (p75NTR) was originally identified as a low affinity receptor for neurotrophins. Recent studies have revealed that p75NTR can promote cell death or survival and modulate neurite outgrowth depending on the operative ligands and co-receptors. Up-regulation and ligand activation of p75NTR have been shown to be involved in neuronal cell death in cultured cells and animal models of neurodegenerative diseases. The levels of proneurotrophins, which bind to p75NTR to promote neuronal death, have been found to be increased in postmortem brains of patients with Alzheimer's disease. Furthermore, there is some evidence for the involvement of this molecule in psychiatric diseases, such as depression and schizophrenia. Mice lacking p75NTR have been shown to have several alterations in central nervous system and cognitive function. Notably, recent progress in genome-based drug discovery has enabled the identification of peptides and non-peptide small molecules targeting p75NTR, which may be potentially beneficial in the treatment of neuropsychiatric diseases. In this review, we focus on recent findings on p75NTR as a therapeutic target for neuropsychiatric diseases. PMID- 20021448 TI - Altered glutamate neurotransmission and behaviour in dementia: evidence from studies of memantine. AB - Behavioural symptoms are a significant problem in Alzheimer's disease (AD). Symptoms including agitation/aggression and psychosis reduce patient quality of life, significantly increase caregiver burden, and often trigger nursing home placement. Underlying changes in the serotonergic, noradrenergic and cholinergic systems have been linked to some behavioural problems, however, the use of antipsychotics in this population has been associated with significant safety concerns. A role for the glutamate system in schizophrenia, as well as in anxiety and depression, has been suggested, and evidence is emerging for a role for dysfunctional glutamate neurotransmission (via N-methyl-D-aspartate (NMDA) receptors) in certain behavioural changes in dementia. For example, the NMDA receptor antagonist, memantine has been shown to improve cognition, function (activities of daily living, ADLs) and, more recently, agitation/aggression, and delusions in AD patients. To date, little information is available regarding the neurochemical basis of agitation/aggression. However, the frontal and cingulate cortices--specifically, the formation of neurofibrillary tangles in glutamatergic pyramidal neurones of these areas--are proposed as regional substrates of these behaviours. Given that memantine displays a favourable tolerability profile, it is relevant to investigate the underlying mechanism linking memantine with the behavioural elements of AD. One hypothesis proposes that memantine corrects dysfunctional glutamatergic neurotransmission in the frontal and cingulate cortices, thereby normalising pathways responsible for causing agitation. An alternative hypothesis is based on the observation that increased tangle formation is associated with agitation, and on recent studies where memantine has been shown to reduce tau phosphorylation via glycogen synthase kinase (GSK)-3 or activation of protein phosphatase (PP)-2A, which might subsequently lead to reduced agitation. PMID- 20021449 TI - Group I metabotropic glutamate receptors: involvement in drug-seeking and drug induced plasticity. AB - L-glutamate is the principal excitatory neurotransmitter at fast synapses in the mammalian central nervous system, and signals though a number of ionotropic and metabotropic receptors. Among the latter are the group I metabotropic glutamate (mGlu1 and mGlu5) receptors that upon activation elevate intracellular calcium levels through activation of the phospholipase C pathway. The role of glutamatergic transmission in both the development of addiction and the phenomenon of relapse that may occur after prolonged abstinence, has come under intense scrutiny in recent times. While both mGlu1 and mGlu5 receptors have been implicated in certain aspects of the addictive state, the exact roles these receptors play in this process is, as yet, unclear. This review will introduce contemporary theories on drug addiction, including neural circuitry, before critically assessing the current body of knowledge on group I metabotropic glutamate receptors in this regard. This will involve an in-depth discussion of the distribution of these receptors in the brain, their presence in neural pathways known or postulated to be involved in addiction and their involvement in drug-related behavioral paradigms. The effect of acute and chronic drug administration on the activity and expression of group I metabotropic glutamate receptors will be investigated, as will the effect these receptors have on behavioral and biochemical responses to drugs of abuse. Finally, there will be a brief discussion on current and future therapeutic applications using our knowledge of these receptors, and the direction that future studies will need to take to close the gaps in our understanding. PMID- 20021450 TI - Biochemical, molecular and epigenetic mechanisms of valproic acid neuroprotection. AB - Valproic acid (VPA, 2-propylpentanoic acid) has been widely used as an antiepileptic drug and for the therapy of bipolar disorders for several years. Its mechanism of action was initially found to be primarily related to neurotransmission and modulation of intracellular pathways. More recently, it emerged as an anti-neoplastic agent as well, by acting on cell growth, differentiation and apoptosis. Here, it mainly exerts its effect by regulating gene expression at the molecular level, through epigenetic mechanisms. In particular, it has been demonstrated the effect of VPA in chromatin remodeling, as VPA directly inhibits histone deacetylases (HDACs) activity. Interestingly, it has been observed that these biochemical and molecular pathways are involved not only in beneficial effect of VPA against epilepsy and malignancies, but they are also responsible for more general neuroprotective mechanisms. In particular, it has been demonstrated that VPA is neuroprotective in several models of neurodegenerative diseases. Moreover, due to the involvement of the VPA-affected mechanisms in complex behaviors, VPA is increasingly used as a psychotherapeutic agent. This review summarizes the more recent data on VPA neuroprotective mechanisms at the biochemical, molecular and epigenetic levels, focusing on both in vitro and in vivo models of neurodegenerative diseases. In particular, attention is paid to mechanisms by which VPA affects neuronal survival/apoptosis and proliferation/differentiation balance, as well as synaptic plasticity, by acting both directly on neurons and indirectly through glial cells. Perspective applications of the VPA neuroprotective potential in human neurodegenerative diseases are discussed, when relevant. PMID- 20021451 TI - Personalised genetic intervention for Duchenne muscular dystrophy: antisense oligomers and exon skipping. AB - Duchenne muscular dystrophy (DMD) arises from protein-truncating mutations in the large dystrophin gene that preclude synthesis of a functional protein that primarily stabilizes muscle fibre membranes. The absence of dystrophin leads to this most common and serious form of childhood muscle-wasting. Since the identification of the dystrophin gene in 1987, cell and gene repair or replacement therapies have been evaluated for DMD treatment and one genetic intervention, exon skipping, is now in clinical trials. Antisense oligomers have been designed to redirect dystrophin splicing patterns so that targeted exons may be removed from a defective dystrophin pre-mRNA to either restore the reading frame of a deletion, or excise an in-frame exon corrupted by a nonsense mutation or micro-insertion/deletion. This review discusses the evolution of oligomer induced exon skipping, including in vitro applications, evaluation of different oligomer chemistries, the treatment of animal models and alternative exon skipping strategies involving viral expression cassettes and ex vivo manipulation of stem cells. The discussion culminates with the current clinical trials and the great challenges that lie ahead. The major obstacle to the implementation of personalised genetic treatments to address the many different mutations that can lead to DMD, are considered to be establishing effective treatments for the different patients and their mutations. Furthermore, the view of regulatory authorities in assessing preclinical data on potentially scores of different but class-specific compounds will be of paramount importance in expediting the clinical application of exon skipping therapy for this serious and relentlessly progressive muscle wasting disease. PMID- 20021452 TI - Pharmacological countermeasures for the acute radiation syndrome. AB - The acute radiation syndrome (ARS) is defined as the signs and symptoms that occur within several months after exposure to ionizing radiation (IR). This syndrome develops after total- or partial-body irradiation at a relatively high dose (above about 1 Gy in humans) and dose rate. Normal tissue injuries induced by IR differ depending on the target organ and cell type. Organs and cells with high sensitivity to radiation include the skin, the hematopoietic system, the gut, the spermatogenic cells and the vascular system. Exposure to IR causes damage to DNA, protein, and lipids in mammalian cells, as well as increased mitochondria-dependent generation of reactive oxygen species (ROS), with subsequent cell cycle checkpoint arrest, apoptosis, and stress-related responses. DNA double strand breaks (DSBs) are a primary lethal lesion induced by IR. The cellular response to damage is complex and relies on simultaneous activation of a number of signaling networks. Among these, the activation of DNA non-homologous end-joining (NHEJ) and homologous recombination (HR), and signaling pathways containing ataxia telangiectasia mutated (ATM), play important roles. The transcription factor NFkappaB has emerged as a pro-survival actor in response to IR in ATM and p53-induced protein with a death domain (PIDD) cascades. Although radiation-induced ARS has been well documented at the clinical level, and mechanistic information is accumulating, successful prophylaxis and treatment for ARS is problematic, even with the use of supportive care and growth factors. There is a pressing need to develop radiation countermeasures that can be used both in the clinic, for small-scale incidents, and outside the clinic, in mass casualty scenarios. In this review we summarize recent information on intracellular and extracellular signaling pathways relevant to radiation countermeasure research. PMID- 20021454 TI - The role of transmembrane segment II in 7TM receptor activation. AB - During the two past decades tremendous effort has been put into uncovering the activation mechanism of 7TM receptors. The majority of such studies have focused on the major binding pocket, comprised of transmembrane segments (TM) -III through -VII, as most non-peptide and peptide ligands, in addition to biogenic amines and retinal a.m.o. bind to residues in this region. Consequently the major helical movements occur here during activation, as described recently in the Global Toggle Switch Model for Family A (also known as rhodopsin-like) members of the 7TM receptors. As a result, the minor binding pocket, comprised of TM-I, -II and, in part, -III and -VII, has received much less attention. With a few exceptions, such as the highly conserved Asp in position II:10/2.50, the residues in this region have generally been considered insignificant with regard to receptor activation. However, accumulating evidence emphasize that this is not the case. In this review, we focus on TM-II with an emphasis on position II:20/2.60, and present data from structure-activity studies on a range of Family A 7TM receptors including chemokine, ghrelin and melanocortin receptors in addition to the orphan EBI2 suggesting that TM-II has important functions for both ligand-dependent and -independent activation of 7TM receptors. PMID- 20021453 TI - The role of the endogenous cannabinoid system in peripheral analgesia. AB - The therapeutic potential of cannabinoids has been studied and investigated through centuries, although many interesting discoveries have emerged from this field in the past decades. Indeed, peripheral analgesic effects of cannabinoids are a new avenue of treatment since they are avoiding the deleterious central side effects of systemic administration. Recently, it has been demonstrated that cannabinoid receptors (more specifically CB(1) and CB(2) receptors) and their endogenous ligands are present at the peripheral level, especially in different layers of skin, and mostly, in the epidermis and dermis. Those findings are reinforcing and confirming the efficacy of peripheral administration of cannabinoids used to alleviate pain in many different animal models. However, many studies have shown that the endocannabinoid system interacts with other receptors and pathways to modulate pain at the peripheral level. Thereof, the main goal of this review is to explain, in a better way, the different interactions regarding the cannabinoid system with other cellular components of its environment, its involvement in the modulation of pain at the peripheral level and, more precisely, in different layers of the skin. PMID- 20021455 TI - E2F1-mediated apoptosis as a target of cancer therapy. AB - The development of cancer depends on the delicate balance between cell proliferation and programmed cell death. p16(INK4a) -cyclinD/cdk4-pRb-E2F1 is a critical pathway regulating cell proliferation in which E2F1 is an important downstream effector. Deregulated E2F1 activity due to the aberrance of the upstream components in this pathway, such as inactivation of Rb or p16, or amplification of cyclinD or cdk4, confers growth advantage to cancer cells and has thus become a hallmark of human tumors. In addition to cell proliferation control, compelling evidence indicates that E2F1 can also induce apoptosis under various cellular contexts regardless of p53 status. Given the frequent inactivation of tumor suppressor pRb and p53 in human cancers, E2F1- induced apoptosis might serve as an additional tumor surveillance mechanism to protect the organism from tumor development. E2F1-mediated apoptosis pathway is therefore emerging as a promising target for therapeutic intervention. Here, we review the recent advances in the understanding of E2F1-mediated apoptosis in cancer cells and discuss how these discoveries may expand our view of E2F1 signaling network and contribute to the development of novel therapeutic strategy. PMID- 20021456 TI - The effects of current therapies on airway remodeling in asthma and new possibilities for treatment and prevention. AB - Airway inflammation, airway remodeling and airway hyperresponsiveness are the fundamental components of pathogenesis that lead to symptoms and lung function changes in asthma. Airway remodeling describes the structural changes to the airways in asthma. The remodeling process involves diverse pathological changes including epithelial metaplasia, subepithelial fibrosis, angiogenesis and smooth muscle thickening. Airway remodeling contributes to irreversible loss of lung function and airway hyperresponsiveness. Remodeling is associated with severe and persistent disease but can also occur early in the course of disease pathogenesis and does not resolve spontaneously. Current asthma therapies, for example corticosteroids, are successful in treating allergic inflammation, an important factor contributing to remodeling, but do not specifically target the remodeling process, and remodeling changes progress despite optimal control of inflammation. Moreover, airway remodeling is not eradicated or prevented despite widespread use of anti-inflammatory treatments. There is limited evidence for the effectiveness of leukotriene inhibitors, phosphodiesterase inhibitors, mast cell tryptase inhibitors, and peroxisome proliferator-activated receptor gamma agonists in the treatment or prevention of remodeling changes. The search for novel therapies that can specifically reverse or prevent airway remodeling is an active area of research. Treatments that may be useful in preventing airway remodeling include those that directly or indirectly target single or multiple components of the airway remodeling process. Identification of novel asthma genes may also allow disease targeting. A better understanding of airway remodeling in asthma will facilitate the development of new treatments for asthma beyond control of symptoms and inflammation. PMID- 20021457 TI - New insights on the possible role of mast cells in aspirin-induced asthma. AB - Nonsteroidal anti-inflammatory drugs (NSAIDs) are major drugs used in the treatment of inflammation and pain in a wide variety of disorders. The best-known mechanism of action of NSAIDs is the inhibition of prostaglandin synthesis as a result of their action on cyclooxygenase (COX) enzymes. However, data have been accumulating through the years indicating that NSAIDs also act on other targets in cell signaling. It has been established that NSAIDs induce anti-inflammatory effects independent of COX. Acetylsalicylic acid (ASA) and other inhibitors of COX induce severe bronchospasms and asthmatic attacks in a significant population of asthmatic patients. The etiology of ASA induced asthma is complex and not fully understood, but most evidence points towards an abnormality of arachidonic acid (AA) metabolism. Since doses of ASA necessary to treat chronic inflammatory diseases appeared much higher than those required to inhibit PG synthesis, COX independent mechanisms of NSAIDs were postulated. Recently, we have shown that NSAIDs induced expression of heat shock proteins specially HSP70. Heat shock proteins (HSPs) are normal intracellular proteins that are produced in greater amounts when cells are subjected to stress or injury. Interestingly, a potential pathogenic role for heat shock proteins in diseases such as autoimmune disease, vascular disease has been reported. Because mast cells have been reported to play a role in the pathogenesis of ASA induced asthma, a link between heat shock proteins and this disease could postulated. In this review, an overview is given on aspirin-induced asthma and the cells and mediators that may play a role therein. Mast cell signaling with regard to interaction with NSAIDs and heat shock proteins (HSPs) and toll-like receptors (TLRs) is further highlighted. PMID- 20021458 TI - Malignant mesothelioma: biology, diagnosis and therapeutic approaches. AB - Malignant mesothelioma (MM) is an aggressive neoplasm of serosal cavities, which is resistant to conventional therapy, with patient survival from presentation of <12 months. MM remains a universally fatal disease of increasing incidence worldwide. Although the main risk factor is asbestos exposure, other factors, Simian virus 40 infection and inheritance of susceptibility genes, likely play a role. Asbestos-related carcinogenic process is primarily based on the interaction between susceptibility (genetic and acquired) and exposure to carcinogenic environmental agents. Asbestos-induced carcinogenesis includes generation of reactive oxygen species, which induce DNA strand breaks and oxidant-induced base modifications to DNA. Persistent oxidative DNA damage can alter signaling cascades, gene expression, induce or arrest transcription, and increase replication errors and genomic instability. The long promotion phase observed in MM pathogenesis and the absence of early symptoms both contribute to late diagnosis of the disease. This results in delayed therapeutic intervention of patients, making the outcome of the disease very grim. There have been several developments in MM management, principally based on early detection, improved diagnosis, development of more effective therapies, and new insights into the pathobiology of the disease. Several programs have been used to screen asbestos exposed individuals for lung and pleural disease. These programs involve annual pulmonary function tests, chest radiography and high resolution computer tomography. Blood tests make screening of target populations an attractive strategy. Many current gene and protein expression studies aim to identify clinically useful biomarkers and new therapeutic targets for improved management of MM. PMID- 20021461 TI - G protein--mediated signaling: same receptor, multiple effectors. AB - The superfamily of G protein coupled receptors (GPCRs) comprises the largest group of cell surface receptors expressed by the human genome. Accordingly, these receptors are the target of a substantial portion of current pharmaceuticals. Over the past few decades there have been many substantial discoveries regarding GPCRs structure and function that have led to the current understanding of the complexity of the signal transduction which these receptors initiate. What was once generally believed to be a simple linear pathway, has become one with manifold bifurcations and multiple regulatory and feedback mechanisms. In the following we review the fundamental ground work upon which this field of research was established and the work that has more recently begun to uncover the complexity of GPCR signaling. The emerging signaling paradigm includes (i) the capacity of one receptor to couple to and initiate pathways through multiple G proteins, (ii) the capability of one G protein to activate many effectors, as well as (iii) the ability of a GPCR to transduce signals through G protein independent pathways. We also briefly touch upon some implications of GPCR oligomerization and discuss signaling cascades of two serotonin receptors, 5 HT(4) and 5-HT(7), whose pathways exemplify the richness and complexity of GPCR signaling mechanisms. PMID- 20021459 TI - Mood-stabilizers target the brain arachidonic acid cascade. AB - Bipolar disorder (BD) is a severe psychiatric illness characterized by recurrent manic and depressive episodes, without a characteristic neuropathology or clear etiology. Drugs effective in BD target many key signaling pathways in animal and cell studies. However, their mode of action in the BD brain remains elusive. In the rat brain, some of the mood stabilizers effective in treating mania (lithium, carbamazepine, valproate) or depression (lamotrigine) in BD are reported to decrease transcription of cytosolic phospholipase A(2) and cyclooxygenase-2 and to reduce levels of AP-2 and NF-kappaB, transcription factors of the two enzymes. The anti-manic drugs also decrease arachidonic acid (AA) turnover in brain phospholipids when given chronically to rats. Thus, drugs effective in BD commonly target AA cascade kinetics as well as AA cascade enzymes and their transcription factors in the rat brain. These studies suggest that of BD is associated with increased AA signaling in the brain. Developing therapeutic agents that suppress brain AA signaling could lead to additional treatments for BD. In this review, we discuss the mechanisms of action of mood stabilizers and the effects of docosahexaenoic acid on AA cascade enzymes in relation to BD. PMID- 20021460 TI - Estrogen receptor neurobiology and its potential for translation into broad spectrum therapeutics for CNS disorders. AB - Estrogens are hormones that modulate a diverse array of effects during development and adulthood. The effects of estrogen are mediated by two estrogen receptor (ER) isotypes, ERalpha and ERbeta, which classically function as transcription factors to modulate specific target gene expression and in addition regulate a growing list of intracellular signaling cascades. These receptors share protein sequence homology and protein-motif organization but have distinct differences in their tissue distribution and binding affinities for their ligands. In the nervous system estrogen has been implicated to play a role in a number of processes which regulate synaptic plasticity including synaptogenesis and neurogenesis. The role for estrogen in a range of neurological and neuropsychiatric diseases is also becoming very apparent. Estrogen is able to regulate processes and behaviours relevant for both Alzheimer's disease and schizophrenia and to modulate neuroendocrine and inflammatory processes important in neuroinflammation, anxiety and depressive disorders as well as chronic pain. We will consider the rationale for estrogen-based therapies for diseases of the nervous system. In particular we will highlight the molecular mechanisms and signal transduction pathways most likely underlying the effects of estrogen in the CNS. PMID- 20021462 TI - Molecular mechanisms of neuronal histamine and its receptors in obesity. AB - Obesity is an important health problem because it is associated with diseases such as type 2 diabetes, hypertension, and hyperlipidemia in metabolic syndrome. The detail molecular mechanisms that underlie obesity have not been fully elucidated, and its therapeutic approach is of general interest. There is increasing evidence that obesity is under control of several factors in the brain and a number of studies have revealed that the brain functions that regulate energy balance play a central role in the development of obesity. Several orexigenic and anorexigenic neuropeptides in the brain are involved in obesity, although their relative contributions are different. The histamine-containing neurons and its receptors are distributed throughout the brain. The results of pharmacological studies revealed that neuronal histamine and its receptors are involved in the regulation of obesity in rodents and humans. In this review, we describe the neuronal histamine and its receptor as a pharmacological molecular target for obesity. PMID- 20021463 TI - Ligand-gated pentameric ion channels, from binding to gating. AB - X-ray structures of molluscan acetylcholine-binding proteins and procaryotic proton-activated ion channels (ELIC and GLIC) enable us to model the ligand binding and activation mechanism of ligand-gated pentameric ion channels. Common versus distinct features can be deduced from the binding of agonists, antagonists and allosteric modulators in subunit interfaces of nicotinic acetylcholine, A type gamma-aminobutyric acid, glycine and 5-HT(3)-type serotonin receptors. Ligand interactions in subunit interfaces elicit conformational waves from the closure of the agonist-binding cavity through binding loops, beta-strands and transmembrane helices to pore gating. PMID- 20021464 TI - Hypoxia and the malignant glioma microenvironment: regulation and implications for therapy. AB - Glioblastoma Multiforme (GBM) tumors are the most common type of brain tumors. These tumors are in general very malignant and can be characterized as rapidly progressive astrocytomas. The pathological characteristics of these tumors are exemplified by an active invasiveness, necrosis and a specialized form of angiogenesis, known as microvascular hyperplasia. These pathological features are thought to be due to tissue hypoxia. Cells that are under hypoxic stress can either develop an adaptive response that includes increasing the rate of glycolysis and angiogenesis or undergo cell death by promoting apoptosis and/or necrosis. The ability of tumor cells to maintain a balance between an adaptation to hypoxia and cell death is regulated by a family of transcription factors called hypoxia-inducing factors (HIF), which are essential for the regulation of the expression of a large number of hypoxia-responsive genes. The hypothesis that tumor hypoxia would facilitate the likelihood of metastases, tumor recurrence, resistance to chemotherapy and radiotherapy and the invasive potential; all of which culminate in a decrease in patient survival. In this review we will summarize the role of hypoxia in GBM with regard to drug therapy and toxicity and attempt to describe the possible interactions between hypoxia and apoptosis. PMID- 20021466 TI - Outcomes of the Fifth International Conference on Pediatric Mechanical Circulatory Support Systems & Pediatric Cardiopulmonary Perfusion. PMID- 20021465 TI - Can colorectal cancer be prevented or treated by oral hormone replacement therapy? AB - Guanylyl cyclase C (GCC) is the receptor specifically expressed by intestinal cells for the paracrine hormones guanylin and uroguanylin and diarrheagenic bacterial heat-stable enterotoxins. This tissue-specific receptor coordinates lineage-dependent regulation of epithelial homeostasis, and its disruption contributes to intestinal tumorigenesis. It coordinates regenerative and metabolic circuits by restricting the cell cycle and proliferation and programming metabolic transitions central to organizing the dynamic crypt-surface axis. Further, mice deficient in GCC signaling are more susceptible to colon cancer induced by Apc mutations or the carcinogen azoxymethane. Moreover, guanylin and uroguanylin are gene products most commonly lost, early, in colon cancer in animals and humans. The role of GCC as a tumor suppressing receptor regulating proliferation and metabolism, together with the universal loss of guanylin and uroguanylin in tumorigenesis, suggests a model in which colorectal cancer is a paracrine hormone deficiency syndrome. In that context, activation of GCC reverses the tumorigenic phenotype by limiting growth of colorectal cancer cells by restricting progression through the G1/S transition and reprogramming metabolic circuits from glycolysis to oxidative phosphorylation, limiting bioenergetic support for rapid proliferation. These observations suggest a pathophysiological hypothesis in which GCC is a lineage-dependent tumor suppressing receptor coordinating proliferative homeostasis whose dysregulation through hormone loss contributes to neoplasia. The correlative therapeutic hypothesis suggests that colorectal cancer is a disease of hormone insufficiency that can be prevented or treated by oral supplementation with GCC ligands. PMID- 20021468 TI - The rewarming index formula for pediatric cardiopulmonary perfusion. AB - Careful rewarming of perfusion blood following cardiopulmonary bypass surgery is critical to a successful outcome, but the optimal rewarming strategy is not clear. The purpose of this study was to derive a formula for a rewarming index (defined as [rewarming time x perfusion flow]/[body weight x body surface area]) that would enable the calculation of the ideal rewarming conditions for pediatric cardiopulmonary perfusion. We retrospectively investigated 220 pediatric cardiopulmonary bypass operations conducted from July 2005 to June 2008 in Okayama University Hospital, Japan. We determined the formula as Phi = (T x Q)/(R x S) = |0.9127P - 0.0152|, where Phi = rewarming index, T = rewarming time (min), Q = perfusion volume (L), R = body weight (kg), S = body surface area (m(2)), and P = temperature gap (right angle). The formula will help those who perform pediatric cardiopulmonary bypass surgery to establish ideal perfusion flow conditions and to control physiological temperature during rewarming. PMID- 20021467 TI - Penn State Hershey--center for pediatric cardiovascular research. PMID- 20021469 TI - Perioperative monitoring of thromboelastograph on hemostasis and therapy for cyanotic infants undergoing complex cardiac surgery. AB - This study investigated features and treatments of perioperative coagulopathies in cyanotic infants with complex congenital heart disease (CCHD). Thirty-six infants with cyanotic CCHD were involved and divided into two groups: In group H (n = 20), hematocrit (HCT) > 54%, and in group L (n = 16), HCT < 54%. Blood was sampled at anesthesia induction (T1), rewarming to 36 degrees C (T2), after heparin neutralization (T3), and 4 h after operation (T4). The hemostatic changes were evaluated by thromboelastograph (TEG). After surgery, group H was treated with fibrinogen-combined platelets (PLT), while group L was treated with PLT only. We observed the effect at T4. At T1, the hemostatic function in group H, deteriorating with the increase of HCT (P < 0.01), was obviously lower than that in group L (P < 0.01), but the PLT function was still complete. In group H, the hemostatic function at T2 decreased with a significant drop of PLT function (P < 0.01) and had little change of functional fibrinogen (Ffg) (P > 0.05). At T3, compared with T2, there were improvements in hemostatic function and Ffg (P < 0.01, respectively) without increase of PLT (P > 0.05) in group H. After therapy, PLT function in both groups restored to T1 level (P > 0.05); Ffg at T4 was significantly better than at T1 (P < 0.01) in group H, but Ffg at T4 with still normal function was lower than at T1 in group L (P < 0.01). Whole hemostatic function at T4 was back to normal and had no differences between two groups. So, we proposed that fibrinogen and PLT transfusion in combination should be better for infants with high HCT CCHD, but PLT alone might be enough for low HCT ones. PMID- 20021470 TI - Pulsatile in vitro simulation of the pediatric univentricular circulation for evaluation of cardiopulmonary assist scenarios. AB - The characteristic depressed hemodynamic state and gradually declining circulatory function in Fontan patients necessitates alternative postoperative management strategies incorporating a system level approach. In this study, the single-ventricle Fontan circulation is modeled by constructing a practical in vitro bench-top pulsatile pediatric flow loop which demonstrates the ability to simulate a wide range of clinical scenarios. The aim of this study is to illustrate the utility of a novel single-ventricle flow loop to study mechanical cardiac assist to Fontan circulation to aid postoperative management and clinical decision-making of single ventricle patients. Two different pediatric ventricular assist devices, Medos and Pediaflow Gen-0, are anastomosed in two nontraditional configurations: systemic venous booster (SVB) and pulmonary arterial booster (PAB). Optimum ventricle assist device strategy is analyzed under normal and pathological (pulmonary hypertension) conditions. Our findings indicate that the Medos ventricular assist device in SVB configuration provided the highest increase in pulmonary (46%) and systemic (90%) venous flow under normal conditions, whereas for the hypertensive condition, highest pulmonary (28%) and systemic (55%) venous flow augmentation were observed for the Pediaflow ventricular assist device inserted as a PAB. We conclude that mechanical cardiac assist in the Fontan circulation effectively results in flow augmentation and introduces various control modalities that can facilitate patient management. Assisted circulation therapies targeting single-ventricle circuits should consider disease state specific physiology and hemodynamics on the optimal configuration decisions. PMID- 20021471 TI - Intravascular mechanical cavopulmonary assistance for patients with failing Fontan physiology. AB - To provide a viable bridge-to-transplant, bridge-to-recovery, or bridge-to surgical reconstruction for patients with failing Fontan physiology, we are developing a collapsible, percutaneously inserted, magnetically levitated axial flow blood pump to support the cavopulmonary circulation in adolescent and adult patients. This unique blood pump will augment pressure and thus flow in the inferior vena cava through the lungs and ameliorate the poor hemodynamics associated with the univentricular circulation. Computational fluid dynamics analyses were performed to create the design of the impeller, the protective cage of filaments, and the set of diffuser blades for our axial flow blood pump. These analyses included the generation of pressure-flow characteristics, scalar stress estimations, and blood damage indexes. A quasi-steady analysis of the diffuser rotation was also completed and indicated an optimal diffuser rotational orientation of approximately 12 degrees. The numerical predictions of the pump performance demonstrated a pressure generation of 2-25 mm Hg for 1-7 L/min over 3000-8000 rpm. Scalar stress values were less than 200 Pa, and fluid residence times were found to be within acceptable ranges being less than 0.25 s. The maximum blood damage index was calculated to be 0.068%. These results support the continued design and development of this cavopulmonary assist device, building upon previous numerical work and experimental prototype testing. PMID- 20021472 TI - Flow distribution during cardiopulmonary bypass in dependency on the outflow cannula positioning. AB - Oxygen deficiency in the right brain is a common problem during cardiopulmonary bypass (CPB). This is linked to an insufficient perfusion of the carotid and vertebral artery. The flow to these vessels is strongly influenced by the outflow cannula position, which is traditionally located in the ascending aorta. Another approach however is to return blood via the right subclavian artery. A computational fluid dynamics (CFD) study was performed for both methods and validated by particle image velocimetry (PIV). A 3-dimensional computer aided design model of the cardiovascular (CV) system was generated from realtime computed tomography and magnetic resonance imaging data. Mesh generation (CFD) and rapid prototyping (PIV) were used for the further model creation. The simulations were performed assuming usual CPB conditions, and the same boundary conditions were applied for the PIV validation. The flow distribution was analyzed for 55 cannula positions inside the aorta and in relation to the distance between the cannula tip and the vertebral artery branch for subclavian cannulation. The study reveals that the Venturi effect due to the cannula jet appears to be the main reason for the loss in cerebral perfusion seen clinically. It provides a PIV-validated CFD method of analyzing the flow distribution in the CV system and can be transferred to other applications. PMID- 20021473 TI - Comparison of pumps and oxygenators with pulsatile and nonpulsatile modes in an infant cardiopulmonary bypass model. AB - As the evidence mounts in favor of pulsatile perfusion during CPB, it is necessary to investigate the effect of circuit components on the quality of pulsatility delivered throughout the circuit. We compared two bloodpumps, the Jostra HL-20 heart-lung machine and the MEDOS DELTASTREAM DP1 Bloodpump, and two oxygenators, the Capiox Baby RX05 and the MEDOS HILITE 800LT, in terms of mean arterial pressure, energy equivalent pressure, surplus hemodynamic energy, total hemodynamic energy, and pressure drop over the oxygenators using a blood analog. The pumps and oxygenators were combined in unique circuits and tested in nonpulsatile and pulsatile modes, at two flow rates (500 and 800 mL/min), and three rotational speed differentials when using the MEDOS DELTASTREAM DP1 Bloodpump for 144 trials in total. The Jostra Roller pump produced some pulsatility in nonpulsatile mode and better pulsatility in pulsatile mode than the MEDOS DP1 Bloodpump at a rotational speed differential of 2500 rpm, but not at 3500 or 4500 rpm. The MEDOS DP1 Bloodpump produced almost no pulsatility in nonpulsatile mode. Pressure drops over the Capiox Baby RX05 were markedly higher, at 92.5 +/- 0.4 mm Hg with the MEDOS DP1 Bloodpump at 800 mL/min and 4500 rpm in pulsatile mode, than those of the MEDOS HILITE 800LT oxygenator, which was 67.0 +/- 0.1 mm Hg at the same settings. These results suggest that careful selection of each circuit component, based on the individual clinical case and component specifics, are necessary to achieve the best quality of pulsatility. PMID- 20021474 TI - Development of mechanical circulatory support devices in China. AB - Myocardial dysfunction leading to low cardiac output syndrome is a common clinical pathophysiological state. Currently, the use of mechanical circulatory support (MCS) is an essential aspect of the treatment of patients with cardiac failure. Several groups in China are engaged in the design and development of MCS devices. These devices can be classified as pulsatile, rotary, and total artificial heart (TAH). There are two types of pulsatile pump, which are driven by air (pneumatic). One of these pumps, the Luo-Ye pump, has been used clinically for short-term support since 1998. The other is a push-plate left ventricular device, which has a variable rate mode. Various rotary devices are classified into axial and centrifugal pumps, depending on the impeller geometry. Most rotary pumps are based on the maglev principle, and some types have been used clinically. Others are still being studied in the laboratory or in animal experiments. Furthermore, certain types of total implantable pump, such as the UJS-III axial pump and the UJS-IV aortic valvo-pump, have been developed. Only one type of TAH has been developed in China. The main constituents of this artificial heart are two axial pumps, two reservoir tanks mimicking the right and left atria, flow meters, two pressure gauges, and a resistance adaptor. Although the development of mechanical assist devices in China is still in a nascent stage, a number of different types of MCS devices are currently being studied. PMID- 20021475 TI - Valve surgery in congenital heart disease. AB - Congenital valve disease (CVD) occurs in isolated form or as part of complex malformations and presents distinct epidemiology, including: young age at onset; high prevalence of associated pathology; history of prior operations; critical clinical presentation. Therefore, multiple interventions are often needed, highlighting the palliative character of CVD surgery. At the same time, long-term survival and satisfactory quality of life expectations must be satisfied in a young, active patient population. The present study analyzes the unique aspects of surgery for CVD by reviewing clinical experience with 565 consecutive patients operated during a 7-year period. Treatment options and outcome are assessed with reference to the ability of respecting the unique demands of patients with CVD. In addition, future developments of CVD management are discussed. PMID- 20021476 TI - Clinical real-time monitoring of gaseous microemboli in pediatric cardiopulmonary bypass. AB - We describe the occurrence and distribution of gaseous microemboli with real-time monitoring in a pediatric cardiopulmonary bypass (CPB) circuit and in the cerebral circulation of patients using the Emboli Detection and Classification (EDAC) system and transcranial Doppler (TCD). Four patients (weights 3.2-13.8 kg) were studied. EDAC monitors were located on the venous line and on the postfilter arterial line to measure gaseous microemboli in the CPB circuit. TCD was used to measure high-intensity transient signals (HITS) in the middle cerebral artery. Before the initiation of CPB, EDAC detected gaseous microemboli in two cases when giving volume through the arterial line. At the initiation of CPB, gross air appeared in the venous line and gaseous microemboli were detected in the arterial line in all patients. EDAC detected a total of 3192-14 699 gaseous microemboli in the arterial line during the whole CPB period, more than 99% of which were smaller than 40 microns. After cessation of CPB, EDAC detected gaseous microemboli in the arterial line in all cases. The TCD detected HITS in two cases (25 and 315), and detected no HITS in two cases. We observed that the venous line acted as a principal source of gaseous microemboli, particularly when using vacuum-assisted venous drainage, and that a significant number of these gaseous microemboli smaller than 40 microns were subsequently transferred to the patient. Using EDAC and TCD together could strengthen the monitoring of gaseous microemboli in the extracorporeal circuit and cerebral circulation. PMID- 20021477 TI - The AJT report: news and issues that affect organ and tissue transplantation. PMID- 20021478 TI - Race and ethnicity in access to and outcomes of liver transplantation: a critical literature review. AB - Racial/ethnic disparities in access to and outcomes of liver transplantation are an important topic given the increasing diversity in the United States. Most reports on this topic predate the advent of allocation based on the model for end stage liver disease (MELD). For many patients with a variety of lethal conditions, liver transplantation is the only effective therapy, signifying the importance of equitable access to care. Racial/ethnic disparities have been described at various steps of the liver transplant process, including liver disease prevalence and treatment, access to a transplant center and its waitlist, receipt of a liver transplant and posttransplant outcomes. The purpose of this minireview is to critically evaluate the published literature on racial/ethnicity based disparities in liver disease prevalence and treatment, transplant center referral, transplant rates and posttransplant outcomes. We identify the shortcomings of previous reports and detail the barriers to completing properly constructed analyses, particularly emphasizing deficits in requisite data and the need for improved study design. Understanding the nature of race/ethnicity-based disparities in liver transplantation is necessary to improve research initiatives, policy design and serves the broader responsibility of providing the highest quality care to all patients with liver disease. PMID- 20021480 TI - Single kidney transplantation from young pediatric donors in the United States. AB - Kidney transplantation (KTX) from small pediatric donors is performed as single or en bloc. Criteria to determine when to split pediatric donor kidneys and transplant as singles are not well established. Data reported to the Scientific Registry of Transplant Recipient for donors <10 yrs from 1995 to 2007 were reviewed (n = 5079). Donors were categorized by weight group by 5 kg increments and solitary (n = 3503) versus en bloc (n = 1576). The primary outcome was overall graft survival. Results were compared as adjusted hazard ratios (aHR) relative to ideal standard criteria donors (SCDs) (defined as age 18-39 without other risk factors), non-ideal SCDs (all other SCDs) and expanded criteria donors (age 50-59 with other risk factors or age >or=60). Single KTX from donors >or= 35 kg conferred a similar risk of graft survival as ideal SCDs. Of donors 10-34 kg, risks of en bloc KTX were similar to ideal and risks of single KTX to non-ideal SCDs; single and en bloc KTXs had 7.9 and 5.2 graft losses per 100 follow-up years, respectively. Single KTX from donors >35 kg are similar to ideal SCDs. Single KTX from donors 10-35 kg are similar to non-ideal SCDs. From a resource perspective, pediatric donors 10-35 kg used as singles offer more cumulative graft years than when used en bloc. PMID- 20021479 TI - CD154 deficiency uncouples allograft CD8+ T-cell effector function from proliferation and inhibits murine airway obliteration. AB - Obliterative bronchiolitis (OB) limits the long-term success of lung transplantation, while T-cell effector mechanisms in this process remain incompletely understood. Using the murine heterotopic tracheal transplant model of obliterative airway disease (OAD) to characterize airway allograft rejection, we previously reported an important role for CD8(+) T cells in OAD. Herein, we studied the role of CD154/CD40 costimulation in the regulation of allospecific CD8(+) T cells, as airway rejection has been reported to be CD154-dependent. Airway allografts from CD154(-/-) recipients had significantly lower day 28 OAD scores compared to wild-type (WT) recipients, and adoptive transfer of CD8(+) T cells from WT recipients, but not CD154(-/-) recipients, were capable of airway rejection in fresh CD154(-/-) allograft recipients. Intragraft CD8(+) T cells from CD154(-/-) mice showed similar expression of the surface markers CD69, CD62L(low) CD44(high) and PD-1, but markedly impaired IFN-gamma and TNF-alpha secretion and granzyme B expression versus WT controls. Unexpectedly, intragraft and systemic CD8(+) T cells from CD154(-/-) recipients demonstrated robust in vivo expansion similar to WT recipients, consistent with an uncoupling of proliferation from effector function. Together, these data suggest that a lack of CD154/CD40 costimulation results in ineffective allospecific priming of CD8(+) T cells required for murine OAD. PMID- 20021481 TI - Apparent remission of a solitary metastatic pulmonary lesion in a liver transplant recipient treated with sorafenib. AB - Hepatocellular carcinoma (HCC) remains a significant disease worldwide and its incidence is expected to increase. In selected patients, liver transplantation offers a 5-year patient survival between 48% and 75%. However, HCC recurrence occurs in approximately 20% of transplant recipients. No therapy has proven efficacious in decreasing the risk of recurrence after transplantation. Sorafenib, a multitargeted tyrosine kinase inhibitor, has been shown to improve survival in patients with advanced HCC that have no history of liver transplantation. We report complete remission of HCC in a 54-year-old man who developed biopsy-proven lung metastasis after liver transplantation treated with sorafenib. PMID- 20021483 TI - Disappearance of gender-related difference in the toxicity of benzotriazole ultraviolet absorber in juvenile rats. AB - 2-(2'-hydroxy-3',5'-di-tert-butylphenyl)benzotriazole (HDBB) is an ultraviolet absorber used in plastic resin products, such as building materials and automobile components. In oral repeated dose toxicity studies using 5- or 6-week old rats, this chemical induced hepatic histopathological changes, such as hypertrophy accompanied with eosinophilic granular changes and focal necrosis of hepatocytes, and male rats showed nearly 25 times higher susceptibility to the toxic effects than females. Castration at approximately 4 weeks of age markedly reduced the sex-related variation in HDBB toxicity, but some difference, less than five times, remained between male and female castrated rats. Following oral HDBB administration to male and female juvenile rats from postnatal days 4-21, such gender-related difference in toxic susceptibility was not detected; therefore, it is speculated that the determinants of susceptibility to HDBB toxicity are differentiated between sexes after weaning. In young rats given HDBB, there was no gender-related difference in plasma HDBB concentration, and no metabolites were detected in the plasma of either sex. HDBB induced lauric acid 12-hydroxylase activity in the liver and this change was more pronounced in males than in females. These findings indicate that HDBB could show hepatic peroxisome proliferation activity, and the difference in the susceptibility of male and female rats to this effect might lead to marked gender-related differences in toxicity. PMID- 20021484 TI - Role of interleukin-15 in the development of mouse olfactory nerve. AB - Interleukin (IL)-15 interacts with components of the IL-2 receptor (R) and exhibits T cell-stimulating activity similar to that of IL-2. In addition, IL-15 is widely expressed in many cell types and tissues, including the central nervous system. We provide evidence of a novel role of IL-15 in olfactory neurogenesis. Both IL-15 and IL-15R alpha were expressed in neuronal precursor cells of the developing olfactory epithelium in mice. Adult IL-15R alpha knockout mice had fewer mature olfactory neurons and proliferating cells than wild-type. Our results suggest that IL-15 plays an important role in regulating cell proliferation in olfactory neurogenesis. PMID- 20021485 TI - Central nervous system congenital anomalies: a prospective neurosurgical observational study from Nigeria. AB - This is a prospective observational study on the profile of central nervous system (CNS) anomalies in a neurosurgical unit in Ikeja, Nigeria. All cases of CNS anomalies seen by the authors in 1 year, January to December 2005, were analyzed. Descriptive variables include some demographics of the patients and their parents; profiles of CNS anomalies, cranial or spinal, neural tube defects (NTD) or others; presence of associated anomalies; and rate of surgical correction of these anomalies. Sixty-one cases were recorded, approximately 64% of our total pediatric neurosurgical output. Forty-one cases were males, 20 females, with a male to female ratio of approximately 2 to 1. The age at presentation ranged from 2 days to 60 months (mean 5.6 months) and the majority of the parents were young (the mean ages of the fathers and mothers were 35 years and 28 years, respectively), and low income earners (70%). The cases comprised cranial (61%) and spinal (39%) anomalies, or 54% of NTD versus others. The cranial lesions were mainly congenital primary hydrocephalus in 44% of cases, encephalocele in 28%, and microcephaly in 17%. The spinal lesions were spinal dysraphism in more than 90%; all in the lumbosacral region. Seventy percent of the NTD had associated cranial or other system anomalies, mainly secondary hydrocephalus in 65%. The surgical correction rate of all of these cases was 38%. NTD were significantly more likely to present early, be associated with other anomalies and undergo surgical correction in this series (P < 0.001). CNS congenital anomalies are under-reported in Nigeria and other developing countries. A call is made for the establishment of nationwide efforts to fully clarify the epidemiology of this silent epidemic. PMID- 20021486 TI - Syndactyly lethal: new mutation with multiple malformations occurring in Sprague Dawley rats. AB - We previously found newborns exhibiting syndactyly of both fore- and hindlimbs in a litter from a pair of Sprague Dawley rats. Continuous breeding of the parental animals yielded pups with the same anomaly in following litters, suggesting that the syndactyly was genetic in origin. In the present study, as all the syndactylous pups died on postnatal day 0, we conducted genetic analyses using 30 phenotypically normal female progeny and the sire. The females were subjected to caesarean section on day 20 of gestation and the fetuses were examined for the phenotypes. The results of the mating experiments suggest that the mutant phenotype is caused by a single autosomal recessive gene at a homozygous condition. As homozygous mutants are lethal at the neonatal stage, the mutant gene was named syndactyly lethal, gene symbol syl. The mutant rats have multiple abnormalities, such as syndactyly, micrognathia, fused/absent/small lung lobes, absent kidney and ureter, small spleen, small uterus, fused phalanges, sternoschisis, absent/detached rib, and splitting/fused/absent/small thoracic vertebra, some of which must be the cause of death on postnatal day 0. This mutant is considered to be useful for investigating the mechanisms and/or pathogenesis of syndactyly, as well as the accompanying malformations. PMID- 20021487 TI - Early embryonic losses in mice induced by diethylstilbestrol. AB - Estrogens cause embryonic lethality and the disturbance of early placental development in mice. Diethylstilbestrol (DES) at 1, 10, or 100 microg/kg was orally administered to Institute of Cancer Research mice on gestational days (GD) 4 through 8, and the uterus and placenta were examined histopathologically on GD 9. Decidua of DES-treated mice showed insufficient development, and the uterine lumen at the implantation site did not effectively minimize. The trophoblast giant cell layer was not separated from the uterine lumen by the decidua capsularis, and hemorrhage from the denuded trophoblast giant cell layer into the uterine lumen was noted at the peripheral part of the decidua basalis. The results of the present study suggest that decidual hypoplasia and subsequent placental hemorrhage causes fetal death due to the administration of DES during the early stage of pregnancy. PMID- 20021488 TI - Cytomegalovirus-associated acute hydramnios treated by amniocentesis and maternal indomethacin. AB - A 22-year-old pregnant woman noticed a rapid increase of abdominal growth, uterine tenderness and irregular contraction, for which she hospitalized at 25 weeks of gestation. An ultrasound examination demonstrated a single fetus with normal anatomy and massive hydramnios. Serial therapeutic amniocentesis was performed for relief of maternal symptoms and indomethacin compress was initiated. Both the maternal and amniotic fluid IgM were positive for cytomegalovirus (CMV). Maternal compress indomethacin was discontinued at 32 weeks. Cesarean section was performed due to fetal distress at 34 weeks of gestation. A female infant was delivered and the neonatal examination was within normal limits with urine culture positive for CMV. At 1 year of age the child was developing normally with normal hearing and no clinical sequelae of intrauterine CMV infection. We postulate that serial and large volume reduction of amniotic fluid by amniocentesis and compress indomethacin in our case interrupted the natural course and provided sufficient time for the fetus to recover from the acute phase of viral infection. PMID- 20021489 TI - Cervical infection secondary to pyriform sinus fistula of branchial origin. AB - Complete third branchial arch anomalies are rare and have been described only in case reports, affecting mainly children and typically presenting as a cervical inflammatory process. Anomalies of the third and fourth branchial apparatus, though rare, usually present as sinuses/incomplete fistulas of pyriform sinus or recurrent suppurative thyroiditis. A 6-year-old girl presented with a small opening on the left side of her anterior neck, which had been present since birth and was associated with recurrent infection. She had no history of incision and drainage of swelling. Computed tomography with contrast injection into the cervical opening revealed a fistulous tract extending from the cervical neck skin to the pyriform fossa. Complete excision of the fistulous tract and left hemithyroidectomy were performed. There was no recurrence at 22 months of follow up. PMID- 20021490 TI - Congenital perineal hernia in a fetus with X monosomy. PMID- 20021493 TI - Developing World Bioethics is turning 10! PMID- 20021491 TI - Tripedus: a form of rachipagus twinning. PMID- 20021494 TI - Capacity building of ethics review committees across Africa based on the results of a comprehensive needs assessment survey. AB - A needs assessment survey of ethics review committees (ERCs) across Africa was conducted in order to establish their major needs and areas of weaknesses in terms of ethical review capacity. The response rate was 84% (31 of 37 targeted committees), and committees surveyed were located in 18 African countries. The majority of the responding committees (61%) have been in existence between 5 and 10 years; approximately 74% of the respondents were institutional committees, with the remainder being either national (6/31) or regional (2/31). In terms of the ethical review process, nine of the 31 committees that responded did not have standard operating procedures (SOPs), and seven of the 22 that did have SOPs had never revised them after their initial development (an average period of three years). Of the 31 committees, 10 operated without any ethical guidelines. Many of the committees (13/30) met once per month, and the number of proposals reviewed annually varied, ranging from five to over 100. All respondents relied on paper based data management and archiving systems. Overall, the survey identified the major constraints on ERCs as lack of office equipment, outdated or lack of SOPs, lack of electronic data management systems, inadequate resources, lack of or insufficient expertise on the committees, and poor recognition of the importance of the role of the committees. Consequently, the authors are addressing the identified needs and weaknesses through the Bill and Melinda Gates Foundation funded capacity building project. The impact of the intervention project will be assessed during and at the end of the four-year longitudinal project. PMID- 20021495 TI - Leishmaniasis entomological field studies: ethical issues. AB - Occupational health remains neglected in developing countries because of competing social, economic and political challenges. Ethical issues in the workplace related to the hazards and risks of becoming infected by Leishmania (Leishmania) mexicana, through the bite of naturally infected sand flies, is another area of concern that has been neglected as well. We report here the results of reviewing two entomological field studies carried out in our research center from 2003 to 2006. Eight students from our School of Biology were invited to catch sand flies. A total of six of the eight (75%) developed a typical clinical picture of Localized Cutaneous Leishmaniasis (LCL) caused by L. (L.) mexicana. In this article we identify the ethical issues related to these kinds of studies and propose some guidelines for conducting them. PMID- 20021498 TI - Vagus nerve stimulation and cardiac asystole. PMID- 20021501 TI - Phenotypic characteristics of novel swine-origin influenza A/California/07/2009 (H1N1) virus. AB - BACKGROUND: The 2009 novel A(H1N1) virus appears to be of swine origin. This strain causing the current outbreaks is a new virus that has not been seen previously either in humans or animals. We have previously reported that viruses causing pandemics or large outbreaks were able to grow at a temperature above the normal physiological range (temperature resistance, non-ts phenotype), were found to be inhibitor resistant and restricted in replication at suboptimal temperature (sensitivity to grow at low temperature, non-ca phenotype). In this study, we performed phenotypic analysis of novel A(H1N1) virus to evaluate its pandemic potential and its suitability for use in developing a live attenuated influenza vaccine. OBJECTIVES: The goal of this study is to identify phenotypic properties of novel A(H1N1) influenza virus. METHODS: A/California/07/2009 (H1N1) swine origin influenza virus was studied in comparison with some influenza A viruses isolated in different years with respect to their ability to grow at non permissive temperatures. We also analyzed its sensitivity to gamma-inhibitors of animal sera and its ability to agglutinate chicken, human and guinea pig erythrocytes. RESULTS: Swine-origin A/California/07/2009 (H1N1) virus was found to be non-ts and inhibitor resistant and was not able to grow at 25 degrees C (non-ca). We did not find any difference in the ability of the hemagglutinin of A/California/07/2009 (H1N1) virus to bind to erythrocytes of different origin. CONCLUSION: The novel swine-origin A(H1N1) virus displays a phenotype typical of the past pandemic and epidemic viruses. This finding suggests that this virus might be a good wild type parental prototype for live vaccine for potential use for controlling pandemic influenza. PMID- 20021502 TI - An ex vivo swine tracheal organ culture for the study of influenza infection. AB - BACKGROUND: The threat posed by swine influenza viruses with potential to transmit from pig populations to other hosts, including humans, requires the development of new experimental systems to study different aspects of influenza infection. Ex vivo organ culture (EVOC) systems have been successfully used in the study of both human and animal respiratory pathogens. OBJECTIVES: We aimed to develop an air interface EVOC using pig tracheas in the study of influenza infection demonstrating that tracheal explants can be effectively maintained in organ culture and support productive influenza infection. METHODS: Tracheal explants were maintained in the air interface EVOC system for 7 days. Histological characteristics were analysed with different staining protocols and co-ordinated ciliary movement on the epithelial surface was evaluated through a bead clearance assay. Explants were infected with a swine H1N1 influenza virus. Influenza infection of epithelial cells was confirmed by immunohistochemistry and viral replication was quantified by plaque assays and real-time RT-PCR. RESULTS: Histological analysis and bead clearance assay showed that the tissue architecture of the explants was maintained for up to 7 days, while ciliary movement exhibited a gradual decrease after 4 days. Challenge with swine H1N1 influenza virus showed that the EVOC tracheal system shows histological changes consistent with in vivo influenza infection and supported productive viral replication over multiple cycles of infection. CONCLUSION: The air interface EVOC system using pig trachea described here constitutes a useful biological tool with a wide range of applications in the study of influenza infection. PMID- 20021503 TI - The effect of age on the pathogenesis of a highly pathogenic avian influenza (HPAI) H5N1 virus in Pekin ducks (Anas platyrhynchos) infected experimentally. AB - BACKGROUND: Highly pathogenic avian influenza (HPAI) H5N1 viruses have recently displayed increased virulence for wild waterfowl. OBJECTIVES: To study the effect of host age on the shedding and tissue dissemination of a HPAI H5N1 virus in infected Pekin ducks. METHODS: Pekin ducks in two age-matched groups (n = 18), 8 and 12 weeks old (wo) were each infected with 10(6) EID(50)/0.1 ml of HPAI A/turkey/Turkey/1/05 (H5N1, clade 2.2). Each day for 5 days, birds were monitored clinically, and cloacal and oropharyngeal swabs collected, before three birds from each group were selected randomly for post-mortem examination. Tissue samples were collected for examination by real-time RT-PCR, histopathology and immunohistochemistry (IHC). RESULTS: Severe clinical signs, including incoordination and torticollis were observed in the 8 wo group resulting in 100% mortality by 4 dpi. Mild clinical signs were observed in the 12 wo group with no mortality. Real-time RT-PCR and IHC results demonstrated the systemic spread of H5N1 virus in birds of both age groups. Higher levels of virus shedding were detected in oropharyngeal swabs than in cloacal swabs, with similar levels of shedding detected in both age groups. Variations in level and temporal dissemination of virus within tissues of older ducks, and the presence of the virus in brain and heart were observed, which coincided with the appearance of clinical signs preceding death in younger birds. CONCLUSIONS: These results are consistent with reports of natural infections of wild waterfowl and poultry possibly indicating an age-related association with dissemination and clinical outcome in ducks following infection with H5N1 HPAI virus. PMID- 20021504 TI - Transmissibility of the 1918 pandemic influenza in Montreal and Winnipeg of Canada. AB - BACKGROUND: The threat of 2009 pandemic influenza A (H1N1) is still causing widespread public concern. A comprehensive understanding of the epidemiology of 1918 pandemic influenza commonly referred to as the Spanish flu may be helpful in offering insight into control strategies for the new pandemic. OBJECTIVE: We explore how the preparedness for a pandemic at the community and individual level impacts the spread of the virus by comparing the transmissibility of the 1918 Spanish flu in two Canadian cities: Montreal and Winnipeg, bearing in mind that each pandemic is unique and the current one may not follow the pattern of the 1918 outbreak. METHODS: The historical epidemiological data obtained for Montreal and Winnipeg in Canada is analyzed to estimate the basic reproduction number which is the most important summary measure of transmission potential of the pandemic. RESULTS: The transmissibility of the 1918 pandemic influenza virus in Winnipeg in the fall of 1918 was found to be much lower than in Montreal based on the estimated reproduction number obtained assuming different serial intervals which are the time between onsets of symptoms in an index case and a secondary case. CONCLUSION: The early preparedness and public health control measures could suggest an explanation for the fact that the number of secondary cases generated by a primary case was significantly reduced in Winnipeg comparing to it in Montreal. PMID- 20021505 TI - Identifying viral infections in vaccinated Chronic Obstructive Pulmonary Disease (COPD) patients using clinical features and inflammatory markers. AB - BACKGROUND: Known inflammatory markers have limited sensitivity and specificity to differentiate viral respiratory tract infections from other causes of acute exacerbation of COPD (AECOPD). To overcome this, we developed a multi-factorial prediction model combining viral symptoms with inflammatory markers. METHODS: Interleukin-6 (IL-6), serum amyloid A (SAA) and viral symptoms were measured in stable COPD and at AECOPD onset and compared with the viral detection rates on multiplex PCR. The predictive accuracy of each measure was assessed using logistic regression and receiver operating characteristics curve (ROC) analysis. RESULTS: There was a total of 33 viruses detected at the onset of 148 AECOPD, the majority 26 (79%) were picornavirus. Viral symptoms with the highest predictive values were rhinorrhoea [Odds ratio (OR) 4.52; 95% CI 1.99-10.29; P < 0.001] and sore throat (OR 2.64; 95% CI 1.14-6.08; P = 0.022), combined the AUC ROC curve was 0.67. At AECOPD onset patients experienced a 1.6-fold increase in IL-6 (P = 0.008) and 4.5-fold increase in SAA (P < 0.001). The addition of IL-6 to the above model significantly improved diagnostic accuracy compared with symptoms alone (AUC ROC 0.80 (P = 0.012). CONCLUSION: The addition of inflammatory markers increases the specificity of a clinical case definition for viral infection, particularly picornavirus infection. PMID- 20021506 TI - Attitudes amongst Australian hospital healthcare workers towards seasonal influenza and vaccination. AB - BACKGROUND: Amongst healthcare workers (HCWs), compliance rates with influenza vaccination are traditionally low. Although a safe and effective vaccine is available, there is little Australian data on reasons for poor compliance, especially amongst allied health and ancillary support staff. METHODS: Cross sectional investigation of a sample of clinical and non-clinical HCWs from two tertiary-referral teaching hospitals in Sydney, Australia was conducted between June 4 and October 19, 2007. The self-administered questionnaire was distributed to hospital personal from 40 different wards and departments. The main outcome measures were personal beliefs about influenza vaccination and self-reported vaccination status. RESULTS: Respondents (n = 1079) were categorized into four main groups by occupation: nurses (47.5%, 512/1079), physicians (26.0%, 281/1079), allied health (15.3%, 165/1079) and ancillary (11.2%, 121/1079). When asked whether they felt the influenza vaccine was safe or effective, 81% (879/1079) and 68% (733/1079), respectively, replied in the affirmative. Participants felt that it was more important to get vaccinated to protect patients (74%, 796/1079) than family (68%, 730/1079) or self-protection (66%, 712/1079). However, only 22% (241/1079) of the HCWs who replied reported receiving the vaccine the year the survey was conducted. CONCLUSIONS: Although HCWs had an adequate level of knowledge towards influenza vaccination, only 22% of them were vaccinated. The approach to improving influenza vaccination rates amongst HCWs and to tackling misconceptions must be multifaceted, adaptable and must evolve regularly to increase coverage. PMID- 20021507 TI - Use and abuse of faecal occult blood tests in an acute hospital inpatient setting. AB - BACKGROUND/AIMS: The role of the faecal occult blood test (FOBT) is untested. The aims of this study were to define the use of FOBT in a general hospital setting and to determine its influence on patient management. METHODS: Case notes and laboratory reports were retrospectively reviewed in all FOBTs performed in 2006 across three acute hospitals, with specific reference to clinical setting, indication, influence over clinical decision-making and management. Both guaiac and immunological tests were performed on all specimens. RESULTS: A total of 330 patients aged 2-104 (mean 74) years, 47% men, had 461 tests performed. A positive result was recorded in one or both tests in 64% of patients. Evidence of dietary restriction was found in only eight (2%) of patients and 218 (66%) patients took one or more medications that could have caused a false positive result. Indications were mostly for overt or suspected gastrointestinal blood loss with or without anaemia and/or iron deficiency, but 5% were for non-bloody diarrhoea and 3% screening for colorectal cancer. Patient care was adversely affected or delayed in 54 patients (16%), mostly because of the result being the stimulus for the decision to refer or not for endoscopy. Only one was considered appropriate as a screening test for colorectal cancer. CONCLUSIONS: The FOBT was applied in clinically inappropriate settings without consideration to confounding issues, and often led to inappropriate clinical decisions with considerable cost to hospital and patient. There is no place for FOBT in an acute hospital setting. PMID- 20021508 TI - Complex lead extraction via subclavian approach using laser technique. AB - Removal of fully intravascular pacing leads may be cumbersome. Herein, we report a technique that may be used to remove intravascular leads using laser technology through a subclavian approach, and discuss the process of lead removal as well as management of potential bleeding from the surgical site. We also address the advantages and pitfalls of this technique as compared to other surgical options. PMID- 20021509 TI - Is concomitant surgery for moderate functional mitral regurgitation indicated during aortic valve replacement for aortic stenosis? A systematic review and evidence-based recommendations. AB - BACKGROUND: Mitral regurgitation (MR) is associated with poor clinical outcomes. Functional MR is often associated with aortic stenosis (AS) and may resolve after aortic valve replacement (AVR). The objective of this study was to derive evidence-based recommendations regarding surgical intervention for moderate functional MR at the time of AVR for AS. METHODS: An exhaustive literature search strategy including Medline, Embase, the Cochrane library, and meeting abstracts was performed. Studies meeting inclusion criteria were critically appraised and data pooled according to accepted meta-analysis techniques. The primary outcome was change in moderate MR after isolated AVR. Secondary outcomes were the impact of functional MR on survival and identifying factors that predict progression of MR, in patients undergoing isolated AVR for AS. RESULTS: Thirteen nonrandomized studies including 2113 patients were reviewed. A total of 268 patients had preoperative moderate functional MR and AS. All studies were appraised as poor methodological quality. After isolated AVR a trend toward improvement in MR was observed. Left ventricular dysfunction, left atrial enlargement, and atrial fibrillation were associated with progression of MR after AVR. However, the impact of residual MR on late survival was not consistent. CONCLUSION: Pooling current evidence provided inconclusive evidence to make clinical practice recommendations for or against routine surgical intervention of moderate MR at the time of AVR for AS. The incidence of this pathology makes further clinical trial studies warranted. PMID- 20021510 TI - Lipomatous hypertrophy of the interatrial septum: a rare cause of right ventricular impairment. AB - We report a case of a 70-year-old woman admitted with symptoms of decompensated heart failure. Magnetic resonance imaging revealed lipomatous hypertrophy of the interatrial septum with partial upper right atrial inflow obstruction, partial obstruction of the right ventricular outflow tract, and excessive accumulation of adipose tissue in the pericardial space. The patient underwent excision of the septal lipomatous mass, which relieved the right ventricular outflow obstruction. PMID- 20021511 TI - Cardiac tamponade as an unusual manifestation of infective endocarditis. AB - Contained rupture of the left ventricle secondary to a myocardial abscess is uncommon. We present a case of infective endocarditis of the aortic valve with an unusual manifestation: a cardiac tamponade due to a pseudoaneurysm of the left ventricle. PMID- 20021512 TI - Neonatal closure of a large coronary-cameral fistula in a case of single coronary artery: the utility of intraoperative angiography. AB - Isolated coronary artery fistula in the newborn is a very rare congenital anomaly. When symptomatic, treatment options include surgical management with ligation and/or patch closure or transcatheter coil embolization. We describe the case of a newborn with symptomatic left coronary-right ventricular fistula, which was managed operatively with off-pump ligation and on-table angiogram for confirmation of fistula obliteration and normal distribution of flow in the coronary artery. We point out the advantages of on-table angiography to evaluate and potentially treat residual structural pathology after cardiac surgery. PMID- 20021513 TI - Distinguishing far-field appendage from local pulmonary vein signal in the left upper pulmonary vein during atrial tachycardia. PMID- 20021514 TI - A novel cryoballoon technique for mapping and isolating pulmonary veins: a feasibility and efficacy study. AB - INTRODUCTION: The study was designed to evaluate the feasibility and efficacy of a simplified cryoballoon technique in which a microcircular catheter was introduced into the central lumen of a cryoballoon catheter for the purpose of recording pulmonary vein (PV) potentials during ablation procedures and without interchanging catheters. METHODS AND RESULTS: A total of 23 consecutive patients with paroxysmal atrial fibrillation (AF) were enrolled. A single transseptal puncture was made and a cryoballoon catheter was inserted into the left atrium. A 6-pole mapping catheter with a 0.035-inch shaft diameter was introduced into the PV through the central lumen of the cryoballoon catheter. In addition to the function as a recording device, the mapping catheter was also used as a "guide wire" during the procedure. A total of 84 PVs (84/92, 91.3%) were completely isolated using this novel cryoballoon technique. In 43 of the 84 veins (51.2%), isolation was observed in real time during the cryoablation; in the remaining 41 veins (48.8%), isolation was confirmed immediately post ablation attempt with the mapping catheter. Procedure time was 152.7 +/- 54.9 minutes, and fluoroscopy time was 33.2 +/- 17.3 minutes. At follow-up (7.4 months, range 2-18 months), 17 (73.9%) patients were free from AF. There was 1 occurrence of phrenic nerve palsy during ablation of a right superior PV, which fully resolved after 1 month. CONCLUSION: The use of a cryoballoon catheter equipped with a 6-pole micromapping catheter inserted through its central lumen for the purpose of mapping and ablation during PV isolation procedures is both feasible and effective. PMID- 20021515 TI - ATP-induced dormant pulmonary veins originating from the carina region after circumferential pulmonary vein isolation of atrial fibrillation. AB - INTRODUCTION: Elimination of transient pulmonary vein recurrences (dormant PVs) induced by an ATP injection and ablation at the PV carina region is an effective strategy for atrial fibrillation (AF) ablation. The relationship between dormant PVs and the PV carina region has not been evaluated. METHODS: A total of 212 consecutive symptomatic AF patients underwent circumferential PV electrical isolation (CPVEI) with a double lasso technique. They were divided into 2 groups in a retrospective review; Group 1: those given an ATP injection during an intravenous isoproterenol infusion after the CPVEI (n = 106), and Group 2: those in which it was not given after the CPVEI (n = 106). Radiofrequency energy was applied at the earliest dormant PV activation site identified using a Lasso catheter on the CPVEI line and then PV carina region if it was ineffective. RESULTS: After a successful PVEI, 54 patients (51%) in Group 1 had PV reconnections during an ATP injection. Acute PVEI sites were observed on the carina region within the CPVEI line in the right PVs (16%) and left PVs (10%). Dormant PVs were reisolated at the carina region in the right PVs (23%) and left PVs (26%). The distribution of the dormant PV sites, except for the RIPV, significantly differed from that of the acute PVEI sites (P < 0.05). Further, AF recurred significantly in the Group 2 patients as compared to those in Group 1 during 16 +/- 6.1 months of follow-up (P < 0.05). CONCLUSION: PV carina region origins may partly be responsible for an acute PVEI and potential recurrences. PMID- 20021516 TI - Atrial fibrillation ablation by aortic retrograde approach using a magnetic navigation system. AB - Stereotaxis remote magnetic navigation system (MNS) (St. Louis, MO, USA) is a new technology with applications in the field of catheter ablation of cardiac arrhythmias. We report a case where the Stereotaxis MNS was successfully used to perform pulmonary vein isolation by a retrograde approach in a patient with atrial fibrillation in whom transseptal access was impossible. PMID- 20021517 TI - Three-dimensional CT overlay in comparison to CartoMerge for pulmonary vein antrum isolation. AB - INTRODUCTION: Three-dimensional (3D) navigation systems are widely used for pulmonary vein antrum isolation (PVAI). To circumvent left atrial (LA) mapping, 3D CT reconstructions of the LA can be superimposed directly (CT overlay) on the fluoroscopy image to guide ablation catheters and to mark ablation sites. METHODS AND RESULTS: Sixty-eight patients (pts) with symptomatic AF refractory to medical therapy were randomly assigned to CT overlay (group 1, n = 38) or CartoMerge (group 2, n = 30). In group 1 registration of the CT image was performed with contrast injections in 2 orthogonal projections. In group 2, visualization of all pulmonary vein (PV) ostia was done by PV angiography, followed by merging of the CT image and the Carto shell. We compared procedural success, procedure time, fluoroscopy time and radiation burden, measured as dose area product (DAP). Baseline characteristics were comparable in both groups. Procedural success, defined as disappearance of PV potentials in all PVs, was achieved in 37/38 (97%) of group 1 patients and 27/30 (90%) patients in group 2 (P = NS). Total procedure time was significantly shorter in group 1 compared to group 2 (129 +/- 34 vs 181 +/- 30 min, P < 0.0001). Although fluoroscopy time tended to be longer in the CT overlay group (47 +/- 16 vs 40 +/- 13 min, P = 0.06), proper use of diaphragmation resulted in comparable radiation values for both groups (DAP 53 +/ 27 vs 56 +/- 35 Gy cm(2), P = 0.76). CONCLUSIONS: CT overlay for PV isolation is feasible and may, in comparison to conventional LA navigation systems, shorten procedural time without increases in radiation burden. PMID- 20021518 TI - An in vitro assessment of acoustic radiation force impulse imaging for visualizing cardiac radiofrequency ablation lesions. AB - INTRODUCTION: Lesion placement and transmurality are critical factors in the success of cardiac transcatheter radiofrequency ablation (RFA) treatments for supraventricular arrhythmias. This study investigated the capabilities of catheter transducer based acoustic radiation force impulse (ARFI) ultrasound imaging for quantifying ablation lesion dimensions. METHODS AND RESULTS: RFA lesions were created in vitro in porcine ventricular myocardium and imaged with an intracardiac ultrasound catheter transducer capable of acquiring spatially registered B-mode and ARFI images. The myocardium was sliced along the imaging plane and photographed. The maximum ARFI-induced displacement images of the lesion were normalized and spatially registered with the photograph by matching the surfaces of the tissue in the B-mode and photographic images. The lesion dimensions determined by a manual segmentation of the photographed lesion based on the visible discoloration of the tissue were compared to automatic segmentations of the ARFI image using 2 different calculated thresholds. ARFI imaging accurately localized and sized the lesions within the myocardium. Differences in the maximum lateral and axial dimensions were statistically below 2 mm and 1 mm, respectively, for the 2 thresholding methods, with mean percent overlap of 68.7 +/- 5.21% and 66.3 +/- 8.4% for the 2 thresholds used. CONCLUSION: ARFI imaging is capable of visualizing myocardial RFA lesion dimensions to within 2 mm in vitro. Visualizing lesions during transcatheter cardiac ablation procedures could improve the success of the treatment by imaging lesion line discontinuity and potentially reducing the required number of ablation lesions and procedure time. PMID- 20021519 TI - Simultaneous electrical and mechanical mapping using 3D cardiac mapping system: novel approach for optimal cardiac resynchronization therapy. AB - Cardiac resynchronization therapy (CRT) restores synchrony in heart failure patients. However, a significant proportion of patients implanted with CRT devices do not realize any benefit from CRT. Placing a left ventricular (LV) lead at the sites of electrical or mechanical delay has been advocated to maximize response to CRT, but there is currently no technique described to measure mechanical delay in real-time. We describe a novel technique that can be used intraoperatively to assess mechanical and electrical activation of the coronary sinus for guidance of LV pacing site optimization during CRT implantation. PMID- 20021520 TI - Endpoints in ablation of paroxysmal atrial fibrillation: when is enough enough? PMID- 20021522 TI - Remote monitoring of implantable cardioverter defibrillators versus quarterly device interrogations in clinic: results from a randomized pilot clinical trial. AB - INTRODUCTION: Remote monitoring is increasingly becoming the new standard of care for implantable cardioverter defibrillator (ICD) follow-up. We sought to determine whether remote monitoring of ICDs improves patient outcomes compared with quarterly device interrogations in clinic. METHODS AND RESULTS: In this single-center pilot clinical trial, adult patients with an ICD were randomly assigned to remote monitoring versus quarterly device interrogations in clinic. The primary endpoint was a composite of cardiovascular hospitalization, emergency room visit for a cardiac cause, and unscheduled visit to the electrophysiology clinic for a device-related issue at 1 year. We also examined health-related quality of life, costs, and patient satisfaction with their ICD care. Of 151 patients enrolled in this trial, 76 were randomized to remote monitoring and 75 to quarterly device interrogations in clinic. There was no significant difference in the primary endpoint (32% in the remote monitoring arm vs 34% in the control arm; P = 0.8), mortality, or cost between the 2 arms. Quality of life and patient satisfaction were significantly better in the control arm than in the remote monitoring arm at 6 months (83 [25th, 75th percentiles 70, 90] vs 75 [50, 85]; P = 0.002 and 88 [75, 100] vs 75 [75, 88]; P = 0.03, respectively), but not at 12 months. CONCLUSION: We showed no significant reduction in cardiac-related resource utilization with remote monitoring of ICDs. However, given the small number of patients in our study, the real clinical and health economics impact of remote monitoring needs to be verified by a large, multicenter, randomized clinical trial. PMID- 20021521 TI - The effect and safety of the antithrombotic therapies in patients with atrial fibrillation and CHADS score 1. AB - BACKGROUND: The revised ACC/AHA/ESC 2006 guideline recommends either aspirin or warfarin for the prevention of ischemic stroke in patients with atrial fibrillation (AF) in CHADS(2) score 1. We hypothesized that warfarin is superior to aspirin therapy for the prevention of stroke without increasing bleeding complication in AF patients with CHADS(2) score 1. METHODS AND RESULTS: Among 1,502 patients (mean 62.4 +/- 13.8 years old, male 65.4%) who were treated for nonvalvular AF without previous stroke, the number of patients with CHADS(2) score 1 was 422 (62.9 +/- 10.7 years old, male 290 [68.7%]) and their antithrombotic therapies were as follows: warfarin (n = 143), aspirin (n = 124), other antiplatelet (n = 45), and no antithrombosis (none: n = 110). We reviewed the incidences of ischemic stroke, mortality, and bleeding complications during the follow-up period. Results were: (1) during 22.3 +/- 17.8 months of follow-up, the incidence of ischemic stroke was significantly lower in warfarin (6 patients, 4.2%, mean international normalized ratio [INR] 2.0 +/- 0.5 IU) than in aspirin (16 patients, 12.9%, P = 0.008) than none (23 patients, 20.9%, P < 0.001) without differences in all-cause mortality. (2) The incidence of major bleeding (decrease in hemoglobin >or=2 g/dL, requiring hospitalization or red blood cell transfusion >or=2 pints) was not different between warfarin (2.1%) and aspirin (0.8%, P = NS), but minor bleeding was more common in warfarin (10.5%) than in aspirin (2.4%, P = 0.007). CONCLUSION: In AF patients with CHADS(2) score 1, warfarin was better to prevent ischemic stroke than aspirin without increasing the incidence of major bleeding complications. However, the incidence of minor bleeding was higher in the warfarin group than the aspirin group. PMID- 20021523 TI - Vagal paroxysmal atrial fibrillation: prevalence and ablation outcome in patients without structural heart disease. AB - INTRODUCTION: The prevalence of vagal and adrenergic atrial fibrillation (AF) and the success rate of pulmonary vein isolation (PVI) are not well defined. We investigated the prevalence of vagal and adrenergic AF and the ablation success rate of antral pulmonary vein isolation (APVI) in patients with these triggers compared with patients with random AF. METHODS AND RESULTS: Two hundred and nine consecutive patients underwent APVI due to symptomatic drug refractory paroxysmal AF. Patients were diagnosed as vagal or adrenergic AF if >90% of AF episodes were related to vagal or adrenergic triggers; otherwise, a diagnosis of random AF was made. Clinical, electrocardiogram (ECG), and Holter follow-up was every 3 months in the first year and every 6 months afterward and for symptoms. Of 209 patients, 57 (27%) had vagal AF, 14 (7%) adrenergic AF, and 138 (66%) random AF. Vagal triggers were sleep (96.4%), postprandial (96.4%), late post-exercise (51%), cold stimulus (20%), coughing (7%), and swallowing (2%). At APVI, 94.3% of patients had isolation of all veins. Twenty-five (12%) patients had a second APVI. At a follow-up of 21 +/- 15 months, the percentage of patients free of AF was 75% in the vagal group, 86% in the adrenergic group, and 82% for random AF (P = 0.51). CONCLUSION: In patients with PAF and no structural heart disease referred for APVI, vagal AF is present in approximately one quarter. APVI is equally effective in patients with vagal AF as in adrenergic and random AF. PMID- 20021524 TI - Health care reform requires renaissance of Flexnerian Academic Medical Centers. PMID- 20021525 TI - Commentary: point. Blood pressure-lowering treatment should be offered to all patients regardless of blood pressure. PMID- 20021526 TI - Commentary: counterpoint. Simple-minded antihypertensive treatment: of assumptions, potpourri, and sausages. PMID- 20021527 TI - Urinary dopamine excretion and renal responses to fenoldopam infusion in blacks and whites. AB - Dopamine is an endogenous natriuretic amine that contributes to the maintenance of sodium homeostasis. Deficiencies in the renal production of dopamine and the action of dopamine on renal tubular receptors have been observed in human hypertension and may contribute to salt sensitivity of blood pressure. Ethnic differences in the sodium-to-dopamine relationship may contribute to the higher prevalence of salt sensitivity in blacks. The authors assessed dopaminergic activity in two studies. In the first, daytime and nighttime excretion of sodium and dopamine were compared in 11 black and 17 white normotensive patients. No racial difference in the rate of sodium or dopamine excretion during either period was observed. In the second study, a graded infusion of the dopamine-1 receptor agonist, fenoldopam, was performed in 14 black and 17 white normotensive patients. There was no racial difference in the natriuretic responses. Previously described lower rates of renal free water clearance and potassium excretion in blacks compared with whites were maintained during fenoldopam infusion, suggesting that dopamine is not a mediator of those differences. The authors conclude that there are no race-related differences in dopamine excretion or activity in normotensive patients. PMID- 20021528 TI - The relationship between flow-mediated dilatation of the brachial artery and intima-media thickness of the carotid artery to Framingham risk scores in older African Americans. AB - The objective of this study was to investigate the relationship of flow-mediated dilatation and intima-media thickness (IMT) with coronary risk in African Americans (AAs). Endothelial dysfunction and IMT of carotid arteries are considered early steps in atherosclerotic disease process and have been used as surrogate markers of subclinical atherosclerosis. Data were collected on 106 AAs with a mean age of 64.0+/-6.6 years. Carotid artery IMT was measured with B-mode ultrasonography, as was brachial artery diameter at rest, during reactive hyperemia, and after nitroglycerin. Percent change in flow-mediated dilatation (%FMD) was defined as 100x(diameter during reactive hyperemia - resting diameter)/resting diameter. Percent change in nitroglycerin-mediated dilatation (%NMD) was defined as 100x(diameter with nitroglycerin-resting diameter)/resting diameter. The Framingham 10-year risk score (FRS) was calculated for each patient using the National Cholesterol Education Program (NCEP) risk score calculator and participants were categorized into 3 groups with FRS as <10%, 10% to 20%, and >20%. Thirty-eight participants had risk scores <10%, 26 had 10% to 20%, and 42 >20%. There was a significant inverse relation between %FMD and FRS (P<.0001) and between %NMD and FRS (P<.001). IMT was not statistically different among the risk groups. Endothelial dysfunction assessed by FMD significantly correlates inversely with FRS in AAs. FMD, an index of arterial compliance, appears to be a sensitive and reliable index of cardiovascular disease. PMID- 20021529 TI - Nebivolol in high-risk, obese African Americans with stage 1 hypertension: effects on blood pressure, vascular compliance, and endothelial function. AB - The authors sought to determine whether nebivolol treatment results in changes in blood pressure (BP), nitric oxide bioavailability, and vascular function in obese African Americans with recently diagnosed stage 1 hypertension. Forty-three obese, hypertensive African Americans (mean BP: systolic, 148.8+/-14.3 mm Hg; diastolic, 90.4+/-8.2 mm Hg) were treated with nebivolol (5-10 mg/d) for 8 weeks. Primary outcomes were change in systolic and diastolic BP and efficacy in reaching normotensive BP. Mean systolic BP decreased by 9.2+/-14 mm Hg (P<.005) and diastolic BP decreased 6.8+/-9 mm Hg (P<.005) with 8 weeks of therapy. Significant improvements were seen in arterial compliance with nebivolol treatment as measured by aortic augmentation index (P<.005) and time to wave reflection (P=.013). Nebivolol treatment improved endothelial function as measured by flow-mediated dilation (P<.005). Levels of erythrocyte cellular superoxide dismutase increased with nebivolol, indirectly suggesting increased bioavailability of nitric oxide (P<.005). Monotherapy with nebivolol in obese, hypertensive African Americans results in significant systolic and diastolic BP reduction by mechanisms that include improved vascular function and compliance. PMID- 20021530 TI - Hypertension in the postpartum woman: clinical update for the hypertension specialist. AB - Hypertension can persist from pregnancy or present de novo in the postpartum period and continue to pose a risk to maternal well-being. These risks are magnified as many patients present after hospital discharge and go unrecognized because of decreased medical surveillance after delivery. Guidelines for the management of postpartum hypertension are lacking, often resulting in imprecise diagnoses and incorrect treatment strategies. As hypertension specialists are called upon to provide advice to obstetricians regarding the management of hypertension in the postpartum period, it becomes important for the hypertension specialist to develop expertise in the evaluation and treatment of hypertensive women during the postpartum period. The purpose of this clinical review article is to provide an approach to the management of postpartum hypertension. PMID- 20021531 TI - Left ventricular thrombus and embolic stroke caused by a functional paraganglioma. PMID- 20021532 TI - Lithium and antihypertensive medication: a potentially dangerous interaction. PMID- 20021533 TI - Largest meta-analysis to date suggests that patients at risk for cardiovascular disease events derive benefit from antihypertensive therapy regardless of baseline blood pressure and to reduce vascular events, lowering blood pressure is more important than choice of antihypertensive drug class. PMID- 20021536 TI - Knowledge, attitude, behavior, and practice of interns toward errors in sphygmomanometer and blood pressure measurement. PMID- 20021537 TI - The metabolic syndrome: a modern plague spread by modern technology. AB - Malnutrition and infectious disease represent the most common health threats facing the developing world. However, increasing technological developments and the expansion of western culture have contributed to the increasing prevalence of the metabolic syndrome. The epidemiologic significance and potential costs to governmental health care systems of an increasing incidence of metabolic syndrome could become high. The role of environmental influences that lead to the development of the metabolic syndrome needs to be explored. Because the metabolic syndrome becomes more common as nations develop, investigations into the ramifications of this disease often come too late. PMID- 20021538 TI - Pathogenesis and pathophysiology of the cardiometabolic syndrome. AB - The cardiometabolic syndrome represents a cluster of metabolic abnormalities that are risk factors for cardiovascular disease. The mechanism(s) responsible for developing the cardiometabolic syndrome is not known, but it is likely that multi organ insulin resistance, which is a common feature of the cardiometabolic syndrome, is involved. Insulin resistance is an important risk factor for type 2 diabetes and can cause vasoconstriction and renal sodium reabsorption, leading to increased blood pressure. Alterations in adipose tissue fatty acid and adipokine metabolism are involved in the pathogenesis of insulin resistance. Excessive rates of fatty acid release into the bloodstream can impair the ability of insulin to stimulate muscle glucose uptake and suppress hepatic glucose production. Noninfectious systemic inflammation associated with adipocyte and adipose tissue macrophage cytokine production can also cause insulin resistance. In addition, increased free fatty acid delivery to the liver can stimulate hepatic very low-density lipoprotein triglyceride production, leading to dyslipidemia. PMID- 20021539 TI - Efficacy of combination drug pulse therapy in maintaining lipid levels in patients intolerant of daily statin use. AB - The objective of this study was to evaluate the efficacy of combination drug pulse therapy in maintaining lipid levels in patients intolerant of a daily dose of statins. Twenty-three patients, previously receiving aggressive statin therapy, were treated twice weekly with rosuvastatin or atorvastatin in different dosages along with ezetimibe as well as daily doses of bile acid sequestrant for a mean period of 4.5 months. The recommended National Cholesterol Education Program Adult Treatment Panel III goals had already been achieved in 78% of patients (n=18) before starting combination pulse therapy. This combination therapy significantly increased high-density lipoprotein cholesterol values by 5.82% (t=2.138, P=.044), while the increases in total cholesterol, low-density lipoprotein cholesterol, triglyceride, and apolipoprotein B levels compared with baseline were not statistically significant. Overall, 3 of 23 patients (13%) discontinued the combination therapy because of muscle-related symptoms over a mean course of 4.5 months of treatment. PMID- 20021540 TI - Association of abdominal adiposity with diabetes and cardiovascular disease in Latin America. AB - The authors explored whether the waist circumference (WC) cutoffs currently proposed to define abdominal obesity (AO) are associated with diabetes and cardiovascular disease (CVD) in Latin America. Primary care physicians in 12 countries were randomly chosen to measure WC and body mass index and record the presence of diabetes and CVD in all consecutive adult patients, consulting them on 2 prespecified half-days. Overall, 70% of 9719 men, and 76% of 18,526 women had AO. Diabetes was reported in 10% of men and 9% of women and CVD in 9% of men and 7% of women. AO was significantly related with diabetes (age-adjusted odds ratio, 1.63 for men and 2.86 for women) and with CVD (odds ratio, 1.41 for men and 1.62 for women). Obesity was also significantly related with diabetes and CVD. Strikingly, abdominal adiposity was very frequent in women with normal body mass index, suggesting that an evidence-based definition of abdominal adiposity in Latin America is needed. PMID- 20021541 TI - Do the current medical and economic times dictate the need for the "polypill"? PMID- 20021543 TI - An analysis of respiratory activity, Q(10), and microbial community composition of soils from high and low tussock sites at Toolik, Alaska. AB - High latitude microbial communities, incurring increased global warming, are a potential major source of respiratory CO2 contributing to an enhanced greenhouse effect. Data on respiration and microbial density are presented for a moist, high tussock site compared with a low, water saturated site. The density of bacteria and eukaryotic microbes was nearly equivalent at both sites and potentially could yield substantial release of respiratory CO2 with continued warming. Respiratory rates for soil from the high site were greater than the low. The Q(10) of 2.4 for the high tussock sample was approximately 1.3 x that of the low site sample (Q(10) of 1.7). PMID- 20021544 TI - Using protistan examples to dispel the myths of intelligent design. AB - In recent years the teaching of the religiously based philosophy of intelligent design (ID) has been proposed as an alternative to modern evolutionary theory. Advocates of ID are largely motivated by their opposition to naturalistic explanations of biological diversity, in accordance with their goal of challenging the philosophy of scientific materialism. Intelligent design has been embraced by a wide variety of creationists who promote highly questionable claims that purport to show the inadequacy of evolutionary theory, which they consider to be a threat to a theistic worldview. We find that examples from protistan biology are well suited for providing evidence of many key evolutionary concepts, and have often been misrepresented or roundly ignored by ID advocates. These include examples of adaptations and radiations that are said to be statistically impossible, as well as examples of speciation both in the laboratory and as documented in the fossil record. Because many biologists may not be familiar with the richness of the protist evolution dataset or with ID-based criticisms of evolution, we provide examples of current ID arguments and specific protistan counter-examples. PMID- 20021545 TI - Introduction: protistan biology, horizontal gene transfer, and common descent uncover faulty logic in intelligent design. PMID- 20021546 TI - Integrating horizontal gene transfer and common descent to depict evolution and contrast it with "common design". AB - Horizontal gene transfer (HGT) and common descent interact in space and time. Because events of HGT co-occur with phylogenetic evolution, it is difficult to depict evolutionary patterns graphically. Tree-like representations of life's diversification are useful, but they ignore the significance of HGT in evolutionary history, particularly of unicellular organisms, ancestors of multicellular life. Here we integrate the reticulated-tree model, ring of life, symbiogenesis whole-organism model, and eliminative pattern pluralism to represent evolution. Using Entamoeba histolytica alcohol dehydrogenase 2 (EhADH2), a bifunctional enzyme in the glycolytic pathway of amoeba, we illustrate how EhADH2 could be the product of both horizontally acquired features from ancestral prokaryotes (i.e. aldehyde dehydrogenase [ALDH] and alcohol dehydrogenase [ADH]), and subsequent functional integration of these enzymes into EhADH2, which is now inherited by amoeba via common descent. Natural selection has driven the evolution of EhADH2 active sites, which require specific amino acids (cysteine 252 in the ALDH domain; histidine 754 in the ADH domain), iron- and NAD(+) as cofactors, and the substrates acetyl-CoA for ALDH and acetaldehyde for ADH. Alternative views invoking "common design" (i.e. the non-naturalistic emergence of major taxa independent from ancestry) to explain the interaction between horizontal and vertical evolution are unfounded. PMID- 20021548 TI - Are you my mother? Kin recognition in the ant Formica fusca. AB - In social insects, workers trade personal reproduction for indirect fitness returns from helping their mother rear collateral kin. Colony membership is generally used as a proxy for kin discrimination, but the question remains whether recognition allows workers to discriminate between kin and nonkin regardless of colony affiliation. We investigated whether workers of the ant Formica fusca can identify their mother when fostered with their mother, their sisters, a hetero-colonial queen or hetero-colonial workers. We found that workers always displayed less aggression towards both their mother and their foster queen, as compared to an unfamiliar hetero-colonial queen. In support of this finding, workers maintain their colony hydrocarbon profile regardless of foster regime, yet show modifications when exposed to different environments. This indicates that recognition entails environmental and genetic components, which allow both discrimination of kin in the absence of prior contact and learning of recognition cues based on group membership. PMID- 20021547 TI - A cancer-causing gene is positively correlated with male aggression in Xiphophorus cortezi. AB - The persistence of seemingly maladaptive genes in organisms challenges evolutionary biological thought. In Xiphophorus fishes, certain melanin patterns form malignant melanomas because of a cancer-causing gene (Xiphophorus melanoma receptor kinase; Xmrk), which arose several millions years ago from unequal meiotic recombination. Xiphophorus melanomas are male biased and induced by androgens however male behaviour and Xmrk genotype has not been investigated. This study found that male X. cortezi with the spotted caudal (Sc) pattern, from which melanomas originate, displayed increased aggression in mirror image trials. Furthermore, Xmrk males (regardless of Sc phenotype) bit and performed more agonistic displays than Xmrk deficient males. Male aggressive response decreased when males viewed their Sc image as compared with their non-Sc image. Collectively, these results indicate that Xmrk males experience a competitive advantage over wild-type males and that intrasexual selection could be an important component in the evolutionary maintenance of this oncogene within Xiphophorus. PMID- 20021549 TI - Local adaptation at the range peripheries of Sitka spruce. AB - High-dispersal rates in heterogeneous environments and historical rapid range expansion can hamper local adaptation; however, we often see clinal variation in high-dispersal tree species. To understand the mechanisms of the species' distribution, we investigated local adaptation and adaptive plasticity in a range wide context in Sitka spruce, a wind-pollinated tree species that has recently expanded its range after glaciations. Phenotypic traits were observed using growth chamber experiments that mimicked temperature and photoperiodic regimes from the limits of the species realized niche. Bud phenology exhibited parallel reaction norms among populations; however, putatively adaptive plasticity and strong divergent selection were seen in bud burst and bud set timing respectively. Natural selection appears to have favoured genotypes that maximize growth rate during available frost-free periods in each environment. We conclude that Sitka spruce has developed local adaptation and adaptive plasticity throughout its range in response to current climatic conditions despite generally high pollen flow and recent range expansion. PMID- 20021550 TI - Relationship between osteology and aquatic locomotion in birds: determining modes of locomotion in extinct Ornithurae. AB - The evolutionary history of aquatic invasion in birds would be incomplete without incorporation of extinct species. We show that aquatic affinities in fossil birds can be inferred by multivariate analysis of skeletal features and locomotion of 245 species of extant birds. Regularized discriminant analyses revealed that measurements of appendicular skeletons successfully separated diving birds from surface swimmers and flyers, while also discriminating among different underwater modes of swimming. The high accuracy of this method allows detection of skeletal characteristics that are indicative of aquatic locomotion and inference of such locomotion in bird species with insufficient behavioural information. Statistical predictions based on the analyses confirm qualitative assessments for both foot propelled (Hesperornithiformes) and wing-propelled (Copepteryx) underwater locomotion in fossil birds. This is the first quantitative inference of underwater modes of swimming in fossil birds, enabling future studies of locomotion in extinct birds and evolutionary transitions among locomotor modes in avian lineage. PMID- 20021551 TI - Genetic variation in flowering phenology and avoidance of seed predation in native populations of Ulex europaeus. AB - The genetic variation in flowering phenology may be an important component of a species' capacity to colonize new environments. In native populations of the invasive species Ulex europaeus, flowering phenology has been shown to be bimodal and related to seed predation. The aim of the present study was to determine if this bimodality has a genetic basis, and to investigate whether the polymorphism in flowering phenology is genetically linked to seed predation, pod production and growth patterns. We set up an experiment raising maternal families in a common garden. Based on mixed analyses of variance and correlations among maternal family means, we found genetic differences between the two main flowering types and confirmed that they reduced seed predation in two different ways: escape in time or predator satiation. We suggest that this polymorphism in strategy may facilitate maintain high genetic diversity for flowering phenology and related life-history traits in native populations of this species, hence providing high evolutionary potential for these traits in invaded areas. PMID- 20021553 TI - Gibberellin and jasmonate crosstalk during stamen development. AB - Gibberellin (GA) and jasmonate (JA) are two types of phytohormones that play important roles during stamen development. For example, Arabidopsis plants deficient in either of GA or JA develop short stamens. An apparent question to ask is whether GA action and JA action during stamen filament development are independent of each other or are in a hierarchy. Recent studies showed that GA modulates the expression of genes essential for JA biosynthesis to promote JA production and high levels of JA will induce the expression of three MYB genes MYB21, MYB24 and MYB57. These three MYB genes are crucial factors for the normal development of stamen filament in Arabidopsis. PMID- 20021554 TI - Identification of microRNAs in wild soybean (Glycine soja). AB - MicroRNAs (miRNAs) play important roles in post-transcriptional gene silencing by directing target mRNA cleavage or translational inhibition. Currently, hundreds of miRNAs have been identified in plants, but no report has been published of wild soybean (Glycine soja Sieb). We constructed a small-RNA library consisting of 2 880 sequences with high quality, in which 1 347 were 19-24 nt in length. By utilizing the miRNA, Rfam and domesticated soybean expressed sequence tag database, we have analyzed and predicted the secondary structure of these small RNAs. As a result, 15 conserved miRNA candidates belonging to eight different families and nine novel miRNA candidates comprising eight families were identified in wild soybean seedlings. All these miRNA candidates were validated by northern blot and the novel candidates expressed in a tissue-specific manner. Furthermore, putative target genes were predicted for novel miRNA candidates and two of them were verified by 5'-rapid amplification of cDNA ends experiments. These results provided useful information for miRNA research in wild soybean and plants. PMID- 20021555 TI - Determination of essential fatty acid composition among mutant lines of Canola (Brassica napus), through high pressure liquid chromatography. AB - The present study aimed to quantify the methyl esters of lenoleic acid (LA), gamma-lenolenic acid (LNA) and oleic acid (OL) in the oil of Brassica napus mutants. Five stable mutants (ROO-75/1, ROO-100/6, ROO-125/12, ROO-125/14, and ROO-125/17) of B. napus cv. 'Rainbow' (P) and three mutants (W97-95/16, W97 0.75/11 and W97-.075/13) of B. napus cv. 'Westar' (P) at M6 stage, exhibiting better yield and yield components, were analyzed for essential fatty acids. The highest seed yield was observed in the mutant (ROO-100/6) followed by ROO-125/14 of Rainbow, that is, 34% and 32% higher than their parent plants, respectively. Westar mutant W97-75/11 also showed 30% higher seed yield than its parent plant. High performance liquid chromatography analysis of the composition of fatty acids indicated that OL was the most dominant fatty acid, ranging from 39.1 to 66.3%; LA was second (15.3-41.6%) and LNA was third (18.1-28.9%). Mutant ROO-125/14 showed higher OL contents than parent (Rainbow). These results are expected to support the approval of ROO-125/14 in the National Uniform Varietal Yield Trials (NUVYT) as a new variety based on high oil quality. PMID- 20021556 TI - Hydrogen sulfide promotes root organogenesis in Ipomoea batatas, Salix matsudana and Glycine max. AB - In this report, we demonstrate that sodium hydrosulfide (NaHS), a hydrogen sulfide (H(2)S) donor, promoted adventitious root formation mediated by auxin and nitric oxide (NO). Application of the H(2)S donor to seedling cuttings of sweet potato (Ipomoea batatas L.) promoted the number and length of adventitious roots in a dose-dependent manner. It was also verified that H(2)S or HS(-) rather than other sulfur-containing components derived from NaHS could be attributed to the stimulation of adventitious root formation. A rapid increase in endogenous H(2)S, indole acetic acid (IAA) and NO were sequentially observed in shoot tips of sweet potato seedlings treated with HaHS. Further investigation showed that H(2)S mediated root formation was alleviated by N-1-naphthylphthalamic acid (NPA), an IAA transport inhibitor, and 2-(4-carboxyphenyl)-4,4,5,5-tetramethylimidazoline-1 oxyl-3-oxide (cPTIO), an NO scavenger. Similar phenomena in H(2)S donor-dependent root organogenesis were observed in both excised willow (Salix matsudana var. tortuosa Vilm) shoots and soybean (Glycine max L.) seedlings. These results indicated that the process of H(2)S-induced adventitious root formation was likely mediated by IAA and NO, and that H(2)S acts upstream of IAA and NO signal transduction pathways. PMID- 20021557 TI - Potato responds to salt stress by increased activity of antioxidant enzymes. AB - To understand the response of potato to salt stress, antioxidant enzyme activities and ion content were analyzed for a sensitive and a tolerant cultivar. Nodal cuttings of the tolerant cultivar, Kennebec, and the sensitive cultivar, Concord, were exposed to media without or with 30, 60, 90 or 120 mmol/L NaCl for 4 weeks. On exposure to NaCl, the length and fresh and dry weight of both shoots and roots of Concord showed greater decrease than those of Kennebec. The decrease in shoot growth was more severe than that of the root for both cultivars. The K(+) content of shoots and roots of both cultivars was reduced in a dose dependent manner by exposure to NaCl; the Na(+) content increased. Activities of ascorbate peroxidase, catalase and glutathione reductase were increased in NaCl exposed shoots of Kennebec; the corresponding activities in NaCl-exposed shoots of Concord were decreased. Roots of both cultivars showed similar changes in the activities of these enzymes on exposure to NaCl. These studies established that enzyme activities in Concord shoots are inversely related to the NaCl concentration, whereas those in Kennebec do not show a dose dependency, which is also the case for the roots of both cultivars. Our findings suggest that an increase in activity of antioxidant enzymes, such as ascorbate peroxidase, catalase and glutathione reductase, can contribute to salt tolerance in Kennebec, a salt resistant cultivar of potato. PMID- 20021558 TI - Drought-stimulated activity of plasma membrane nicotinamide adenine dinucleotide phosphate oxidase and its catalytic properties in rice. AB - The activity of plasma membrane (PM) nicotinamide adenine dinucleotide phosphate (NADPH) oxidase and its catalytic properties in rice was investigated under drought stress conditions. Drought stress led to decreased leaf relative water content (RWC) and, as a result of drought-induced oxidative stress, the activities of antioxidant enzymes increased significantly. More interestingly, the intensity of applied water stress was correlated with increased production of H2O2 and O2 (-) and elevated activity of PM NADPH oxidase, a key enzyme of reactive oxygen species generation in plants. Histochemical analyses also revealed increased H2O2 and O2 (-) production in drought-stressed leaves. Application of diphenylene iodonium (DPI), an inhibitor of PM NADPH oxidase, did not alleviate drought-induced production of H2O2 and O2 (-). Catalysis experiments indicated that the rice PM NADPH oxidase was partially flavin dependent. The pH and temperature optima for this enzyme were 9.8 and 40 degrees C, respectively. In addition, drought stress enhanced the activity under alkaline pH and high temperature conditions. These results suggest that a complex regulatory mechanism, associated with the NADPH oxidase-H2O2 system, is involved in the response of rice to drought stress. PMID- 20021559 TI - Development of a core set from a large rice collection using a modified heuristic algorithm to retain maximum diversity. AB - A new heuristic approach was undertaken for the establishment of a core set for the diversity research of rice. As a result, 107 entries were selected from the 10 368 characterized accessions. The core set derived using this new approach provided a good representation of the characterized accessions present in the entire collection. No significant differences for the mean, range, standard deviation and coefficient of variation of each trait were observed between the core and existing collections. We also compared the diversity of core sets established using this Heuristic Core Collection (HCC) approach with those of core sets established using the conventional clustering methods. This modified heuristic algorithm can also be used to select genotype data with allelic richness and reduced redundancy, and to facilitate management and use of large collections of plant genetic resources in a more efficient way. PMID- 20021560 TI - Association analysis of the amino acid contents in rice. AB - The main objective of the present study was to identify simple sequence repeat (SSR) markers associated with the amino acid content of rice (Oryza sativa L.). SSR markers were selected by prescreening for the relationship to amino acid content. Eighty-four rice landrace accessions from Korea were evaluated for 16 kinds of amino acids in brown rice and genotyped with 25 SSR markers. Analysis of population structure revealed four subgroups in the population. Linkage disequilibrium (LD) patterns and distributions are of fundamental importance for genome-wide mapping associations. The mean r(2) value for all intrachromosomal loci pairs was 0.033. LD between linked markers decreased with distance. Marker trait associations were investigated using the unified mixed-model approach, considering both population structure (Q) and kinship (K). A total of 42 marker trait associations with amino acids (P < 0.05) were identified using 15 different SSR markers covering three chromosomes and explaining more than 40% of the total variation. These results suggest that association analysis in rice is a viable alternative to quantitative trait loci mapping and should help rice breeders develop strategies for improving rice quality. PMID- 20021561 TI - Performance of hybrids between weedy rice and insect-resistant transgenic rice under field experiments: implication for environmental biosafety assessment. AB - Transgene escape from genetically modified (GM) rice into weedy rice via gene flow may cause undesired environmental consequences. Estimating the field performance of crop-weed hybrids will facilitate our understanding of potential introgression of crop genes (including transgenes) into weedy rice populations, allowing for effective biosafety assessment. Comparative studies of three weedy rice strains and their hybrids with two GM rice lines containing different insect resistance transgenes (CpTI or Bt/CpTI) indicated an enhanced relative performance of the crop-weed hybrids, with taller plants, more tillers, panicles, and spikelets per plant, as well as higher 1 000-seed weight, compared with the weedy rice parents, although the hybrids produced less filled seeds per plant than their weedy parents. Seeds from the F(1) hybrids had higher germination rates and produced more seedlings than the weedy parents, which correlated positively with 1 000-seed weight. The crop-weed hybrids demonstrated a generally enhanced relative performance than their weedy rice parents in our field experiments. These findings indicate that transgenes from GM rice can persist to and introgress into weedy rice populations through recurrent crop-to-weed gene flow with the aid of slightly increased relative fitness in F(1) hybrids. PMID- 20021564 TI - Requirement for small side chain residues within the GxGD-motif of presenilin for gamma-secretase substrate cleavage. AB - gamma-Secretase is a pivotal intramembrane-cleaving protease complex and important drug target for Alzheimer's disease. The protease not only releases small peptides, such as the amyloid-beta peptide, which drives Alzheimer's disease pathogenesis, but also intracellular domains, which can have critical functions in nuclear signaling. Unlike typical aspartyl proteases, gamma secretase contains a non-classical GxGD active site motif in its catalytic subunit presenilin (PS) 1 or PS2. It is not known whether both glycines are of similar functional relevance and why the glycine residues are invariant elements of the motif. Here we identify the N-terminal glycine of the GxGD motif in PS1, G382, as a critical residue of the active site domain of gamma-secretase. Substitution of G382 by a number of different amino acids abrogated gamma secretase activity. Only the smallest possible G382A substitution allowed substantial gamma-secretase activity. Depending on the substrate, however, the presence of G382 could become even an absolute functional requirement of gamma secretase. Very similar results were obtained for the C-terminal glycine residue (G384) of the GxGD motif. Our data thus identify a requirement for small side chain residues in the active site domain of gamma-secretase and suggest that the glycines of the GxGD motif could be evolutionary conserved to allow cleavage of all possible gamma-secretase substrates, including those, which are highly sensitive to minimal alteration of the PS active site domain. These findings broaden our understanding of gamma-secretase substrate recognition and cleavage, which may prove crucial for therapeutic targeting of the enzyme. PMID- 20021565 TI - Ethanol induction of steroidogenesis in rat adrenal and brain is dependent upon pituitary ACTH release and de novo adrenal StAR synthesis. AB - The mechanisms of ethanol actions that produce its behavioral sequelae involve the synthesis of potent GABAergic neuroactive steroids, specifically the GABAergic metabolites of progesterone, (3alpha,5alpha)-3-hydroxypregnan-20-one (3alpha,5alpha-THP), and deoxycorticosterone, (3alpha,5alpha)-3,21 dihydroxypregnan-20-one. We investigated the mechanisms that underlie the effect of ethanol on adrenal steroidogenesis. We found that ethanol effects on plasma pregnenolone, progesterone, 3alpha,5alpha-THP and cortical 3alpha,5alpha-THP are highly correlated, exhibit a threshold of 1.5 g/kg, but show no dose dependence. Ethanol increases plasma adrenocorticotropic hormone (ACTH), adrenal steroidogenic acute regulatory protein (StAR), and adrenal StAR phosphorylation, but does not alter levels of other adrenal cholesterol transporters. The inhibition of ACTH release, de novo adrenal StAR synthesis or cytochrome P450 side chain cleavage activity prevents ethanol-induced increases in GABAergic steroids in plasma and brain. ACTH release and de novo StAR synthesis are independently regulated following ethanol administration and both are necessary, but not sufficient, for ethanol-induced elevation of plasma and brain neuroactive steroids. As GABAergic steroids contribute to ethanol actions and ethanol sensitivity, the mechanisms of this effect of ethanol may be important factors that contribute to the behavioral actions of ethanol and risk for alcohol abuse disorders. PMID- 20021566 TI - D3 dopamine receptor signals to activation of phospholipase D through a complex with Rho. AB - Dopamine acts through a family of G protein-coupled receptors to exert its myriad effects. The D3 Dopamine receptor is one member of the D2-like dopamine receptors. We have previously demonstrated in human embryonic kidney (HEK293) cells that D3 receptor stimulation of phospholipase D (PLD) activity is pertussis toxin insensitive [Everett and Senogles. Neurosci. Lett. 371 (2004), 34]. We hypothesized that a low molecular weight G protein was involved in the agonist mediated activation of PLD. When the D3 receptor was coexpressed with RhoA in HEK293 cells, agonist-induced stimulation of PLD activity was increased. However, co-expression of Rac or Cdc42 with the D3 receptor did not change the PLD activity. As well, expression of a dominant-negative construct of RhoA, N19 Rho completely ablated D3 receptor-mediated PLD activation, when co-expressed with the D3 receptor in HEK293 cells. In contrast, expression of dominant-negative constructs of Rac or Cdc42 had no effect. Treatment of HEK293 cells transfected with the D3 receptor and treated with a D3 preferring agonist R+ hydroxy(dipropylamino)tetralin hydrobromide, results in an agonist-induced physical complex of D3 receptor and either endogenous Rho or transfected hemaglutinin (HA)-RhoA that can be detected by immunoprecipitation and western blotting. Treatment of cells transfected with D3 receptor with R+ hydroxy(dipropylamino)tetralin hydrobromide also results in agonist-dependent Rho activation, as measured by a Rho effector pull-down assay. The data suggest that D3 receptor/RhoA association and activation is necessary for D3 receptor-mediated PLD activation. PMID- 20021568 TI - Improving fatigue assessment in immune-mediated neuropathies: the modified Rasch built fatigue severity scale. AB - Fatigue is a major disabling complaint in patients with immune-mediated neuropathies (IN). The 9-item fatigue severity scale (FSS) has been used to assess fatigue in these conditions, despite having limitations due to its classic ordinal construct. The aim was to improve fatigue assessment in IN through evaluation of the FSS using a modern clinimetric approach [Rasch unidimensional measurement model (RUMM2020)]. Included were 192 stable patients with Guillain Barre syndrome (GBS), chronic inflammatory demyelinating polyradiculoneuropathy (CIDP) or polyneuropathy associated with monoclonal gammopathy of undetermined significance (MGUSP). The obtained FSS data were exposed to RUMM2020 model to investigate whether this scale would meet its expectations. Also, reliability and validity studies were performed. The original FSS did not meet the Rasch model expectations, primarily based on two misfitting items, one of these also showing bias towards the factor 'walking independent.' After removing these two items and collapsing the original 7-point Likert options to 4-point response categories for the remaining items, we succeeded in constructing a 7-item Rasch-built scale that fulfilled all requirements of unidimensionality, linearity, and rating scale model. Good reliability and validity were also obtained for the modified FSS scale. In conclusion, a 7-item linearly weighted Rasch-built modified FSS is presented for more proper assessment of fatigue in future studies in patients with immune-mediated neuropathies. PMID- 20021567 TI - Hyperlipidemia: a new therapeutic target for diabetic neuropathy. AB - Emerging data establish dyslipidemia as a significant contributor to the development of diabetic neuropathy. In this review, we discuss how separate metabolic imbalances, including hyperglycemia and hyperlipidemia, converge on mechanisms leading to oxidative stress in dorsal root ganglia (DRG) sensory neurons. We conclude with suggestions for novel therapeutic strategies to prevent or reverse diabetes-induced nerve degeneration. PMID- 20021569 TI - Neuropathic pain is associated with increased nodal persistent Na(+) currents in human diabetic neuropathy. AB - Peripheral nerve injury alters function and expression of voltage gated Na(+) channels on the axolemma, leading to ectopic firing and neuropathic pain/paresthesia. Hyperglycemia also affects nodal Na(+) currents, presumably due to activation of polyol pathway and impaired Na(+)-K(+) pump. We investigated changes in nodal Na(+) currents in peripheral sensory axons and their relation with pain in human diabetic neuropathy. Latent addition using computerized threshold tracking was used to estimate nodal persistent Na(+) currents in radial sensory axons of 81 diabetic patients. Of these, 36 (44%) had chronic neuropathic pain and severe paresthesia. Compared to patients without pain, those with pain had greater nodal Na(+) currents (p = 0.001), smaller amplitudes of sensory nerve action potentials (SNAP) (p = 0.0003), and lower hemoglobin A1c levels (p = 0.006). Higher axonal Na(+) conductance was associated with smaller SNAP amplitudes (p = 0.03) and lower hemoglobin A1c levels (p = 0.008). These results suggest that development of neuropathic pain depends on axonal hyperexcitability due to increased nodal Na(+) currents associated with structural changes, but the currents could also be affected by the state of glycemic control. Our findings support the view that altered Na(+) channels could be responsible for neuropathic pain/paresthesia in diabetic neuropathy. PMID- 20021570 TI - PEMF fails to enhance nerve regeneration after sciatic nerve crush lesion. AB - The use of electromagnetic fields has been reported to enhance peripheral nerve regeneration. This study aimed to identify the effects of a prolonged protocol of low-frequency pulsed electromagnetic field (PEMF) on peripheral nerve regeneration. Thirty-four male Swiss mice (Mus musculus) were divided into PEMF (n = 17) and control (n = 17) groups. All animals underwent a unilateral sciatic crush lesion, and the PEMF group was exposed to a 72-Hz, 2-G electromagnetic field for 30 min, five days a week, for three weeks. Functional analysis was carried out weekly. After three weeks, the animals were euthanized, and histological, morphometric, oxidative stress, and TGF-beta1 analyses were performed. Functional analysis showed no differences between the groups. Histological appearance was similar between PEMF and control nerves. Morphometric assessment showed that the PEMF nerves trended toward decreased regeneration. The levels of free radicals were more pronounced in PEMF nerves, but were not associated with an increase in the content of the TGF-beta1/Smad signaling pathway. Prolonged PEMF regimen leads to delayed histological peripheral nerve regeneration and increased oxidative stress but no loss of function recovery. PMID- 20021571 TI - C-fiber axon reflex flare size correlates with epidermal nerve fiber density in human skin biopsies. AB - The size of the neurogenic axon reflex flare (ARFS) has been proposed to serve as a non-invasive measure of C-fiber neuropathies. This idea is based on the observation that ARFS is often reduced in patients with small-fiber neuropathies. In this study, we compared ARFS and electrically evoked axon reflex sweating with intraepidermal nerve fiber density (IENF) in patients with peripheral neuropathy in order to validate these methods against an objective standard method of diagnosing small-fiber neuropathy. ARFS was significantly correlated with IENF, while axon reflex sweating was not correlated to IENF. We conclude that measurement of ARFS is a potential objective non-invasive diagnostic tool for analysis of C-fiber function in patients with small-fiber neuropathies. PMID- 20021572 TI - Effects of transient receptor potential (TRP) channel agonists and antagonists on slowly adapting type II mechanoreceptors in the rat sinus hair follicle. AB - The possible functional role of transient receptor potential (TRP) channels was investigated by testing various TRP agonists and antagonists in an isolated rat sinus hair follicle preparation. Extracellular recordings from slowly adapting type II mechanoreceptor units were made. The antagonist capsazepine depressed spontaneous and mechanically evoked activity, with an IC(50) of 82 microM. In one third of units, capsazepine caused a selective depression of mechanically evoked firing, such that the existing spontaneous firing was interrupted by an absence of activity during the mechanical stimulus. The broad spectrum TRP blocker ruthenium red (30 microM) had inconsistent effects, although in some units a delayed onset (following wash) bursting and paroxysmal firing ensued. The agonist icilin (50-100 microM) had an excitatory effect on spontaneous firing, and (-) menthol (200 microM) had inconsistent effects. Cinnamaldehyde (1-2 mM) depressed all types of activity equally, mechanically evoked and spontaneous. Camphor (0.5 2 mM) also depressed all types of activity, although it had a preferential effect on spontaneous activity. Capsaicin (1-10 microM) and allyl isothiocyanate (50-100 microM) had no clear effects. These results rule out any role for TRPA1 and TRPV1 channels in mechanotransduction processes of slowly adapting type II mechanoreceptors. PMID- 20021573 TI - Recurrences, vaccinations and long-term symptoms in GBS and CIDP. AB - We determined the frequency of recurrent Guillain-Barre syndrome (GBS), whether vaccinations led to recurrences of GBS or an increase of disability in chronic inflammatory demyelinating polyradiculoneuropathy (CIDP) and we assessed the prevalence of pain, fatigue and the impact on quality of life after GBS and CIDP. Additionally, we assessed the presence of common auto-immune disorders. Four hundred and sixty-one members of the Dutch society of neuromuscular disorders received a questionnaire. Two hundred and forty-five GBS and seventy-six CIDP patients were included (response rate 70%). Nine patients had a confirmed recurrent GBS, and two patients had experienced both GBS and CIDP. Common auto immune diseases were reported in 9% of GBS and 5% of CIDP patients. None of the 106 GBS patients who received a flu vaccination (range 1-37 times, total 775 vaccinations) reported a recurrence thereafter. Five out of twenty-four CIDP patients who received a flu vaccination (range 1-17 times) reported an increase in symptoms. Pain or severe fatigue was reported in about 70% of patients after the diagnosis of GBS (median 10 years) or after onset of CIDP (median 6 years), and quality of life was significantly reduced. Flu vaccinations seem relatively safe. GBS and CIDP patients often experience pain, fatigue and a reduced quality of life for many years after the diagnosis. PMID- 20021574 TI - Guillain-Barre syndrome mimicking brain death. PMID- 20021575 TI - Roles and rewards of journal peer reviewers. PMID- 20021576 TI - Relationship between assertiveness and burnout among nurse managers. AB - AIM: We aimed to clarify the relationship between assertiveness and burnout among nurse managers at university hospitals. METHODS: The directors at three university hospitals agreed to cooperate with our study. During a one-month period from May to June 2007, a self-administered questionnaire was distributed to 203 nurse managers (head and sub-head nurses). The Japanese version of the Rathus Assertiveness Schedule (J-RAS) and the Japanese version of the Maslach Burnout Inventory (MBI) were used as scales. Burnout was operationally defined as a total MBI score in the highest tertile. RESULTS: Valid responses were obtained from 172 nurse managers. The mean J-RAS score of the burnout group (-14.3) was significantly lower than that of the non-burnout group (-3.3). Responses about work experience and age showed no significant group difference. Total MBI score was inversely correlated with J-RAS score (R = -0.30, P < 0.01). Multiple logistic regression analyses indicated a decrease in the risk of burnout by 26% (0.74 times) for every 10 point increase in the J-RAS score, and by 60% (0.40 times) for greater satisfaction with own care provision. CONCLUSIONS: The results suggest that increasing assertiveness and satisfaction with own care provision contributes to preventing burnout among Japanese nurse managers. PMID- 20021577 TI - Development of a community health promotion center based on the World Health Organization's Ottawa Charter health promotion strategies. AB - AIM: To describe the development process of nursing school-led community health promotion centers (CHPC) to improve the health of the surrounding communities. METHODS: This study design was a research and development study. (i) Assessment of health needs by interviewing 359 people in the community to select health programs for the community health promotion center. (ii) Five health promotion strategies from the Ottawa Charter were applied to develop the community health promotion center for a city community. RESULTS: (i) The people in the community had higher socioeconomic status levels and better health behaviors compared to the general Korean population, and they also listed chronic health problem management as their first priority health service. (ii) Development of the community health promotion center was done based on the five World Health Organization's Ottawa Charter Health Promotion Strategies: build healthy public policy, create supportive environments, strengthen community actions, develop personal skills, and reorient health services. CONCLUSIONS: The present study showed that the WHO's five Ottawa Charter Health Promotion Strategies were useful for developing health promotion centers in the community. PMID- 20021578 TI - Education to promote verbal communication by caregivers in geriatric care facilities. AB - AIM: Our previous study divided the verbal communication between caregivers and elderly residents at geriatric care facilities into Type I communication (to elicit activities of daily living) and Type II communication (conversation that occurs in normal social life) and found that Type II communication promotes utterances by elderly residents. This study conducted an education intervention to promote Type II talking by caregivers and evaluated the results. METHODS: At three geriatric care facilities, 243 caregivers who might care for 36 elderly residents experienced training involving lectures and group discussion to understand the importance of Type II talking and how to apply it to their daily work. A statistical comparison was applied to the changes in Type II talking duration from before the intervention, 1 week after the intervention, and 3 months after the intervention to evaluate the effect of the educational intervention. RESULTS: At two facilities, the Type II talking duration increased significantly from before the educational intervention to 1 week after the intervention and remained higher after 3 months. However, the educational intervention's effect was not clear at one facility. There was no significant difference in the elderly persons' total utterance duration, but it increased from before the intervention to 1 week after the intervention. CONCLUSION: After the educational intervention, the amount of Type II talking by the caregivers increased significantly 1 week after the intervention for two facilities, but although the amount of Type II talking was higher at 3 months than before the intervention, it was not as high as 1 week after the intervention. PMID- 20021579 TI - Depression and hopelessness in Turkish patients with cancer undergoing chemotherapy. AB - AIM: This study aimed to determine the levels of depression and hopelessness of patients receiving chemotherapy. Through knowledge of the levels of hopelessness and depression in such patients, this study could contribute to the planning of nursing interventions. METHODS: The study involved 101 patients with cancer who presented to the outpatient unit of a medical oncology clinic to receive outpatient chemotherapy between January and March 2006. Data on the patients' sociodemographic features, as well as their scores on the Beck Hopelessness Scale and the Beck Depression Inventory, were obtained. RESULTS: The patients' mean total depression score was 16.0 +/- 8.3 and their mean hopelessness score was 6.9 +/- 3.4. There was a statistically significant positive relationship between depression and hopelessness. CONCLUSION: The results indicated that depression and hopelessness were strongly and positively correlated. PMID- 20021580 TI - Frequency and type of chronic pain care approaches used for elderly residents in Japan and the factors influencing these approaches. AB - AIM: To assess the frequency at which various chronic pain care (CPC) approaches were used while managing older residents of the Health Service Facilities for the Elderly Requiring Care (HSFERC) in Japan and to assess the factors related to nurses and care workers that influence this care. METHODS: A descriptive study design was used. The population comprised 31 nurses, 92 care workers, and 18 residents with chronic pain in eight HSFERC centers located in three provincial cities in Japan. A questionnaire was formulated by using the data collected by a literature review to assess the frequencies at which various CPC approaches were applied and the factors that might influence this care. RESULTS: The most frequently preferred CPC approaches were gentle handling and support while providing daily care, listening attentively, and providing a recreational activity. The factors that affected the provision of CPC were the qualifications, years of experience of aged care, and experience of studying about chronic pain. The nurses tended to have a misconception regarding the manner in which the residents complained of pain and their pain sensitivity. Furthermore, organizational strategies for pain management were not reported by the nurses and care workers. CONCLUSIONS: In order to provide effective and active CPC, ongoing education about pain and cooperation between nurses and care workers to manage residents' pain are highly recommended. PMID- 20021581 TI - Experiences of difficulty that patients with cancer faced in the learning process of progressive muscle relaxation. AB - AIM: The purpose of this study was to clarify the experiences of difficulty that patients with cancer faced in the learning process of progressive muscle relaxation (PMR) and to consider which kind of PMR guidance should be given to weak patients with cancer. METHODS: The participants were instructed to practise PMR for 2 weeks when they were in good condition. The interview was conducted 1 week and 2 weeks after the PMR intervention. The 21 participants who experienced difficulty were analyzed. The analysis was completed by using the content analysis method. RESULTS: Fifteen patients practised PMR by themselves one-to-two times per day. However, six patients practised PMR three-to-four times per week when they were in good condition. The interview answers fell into two categories: (i) experiences of difficulty that patients encountered in the learning process; and (ii) experiences in which patients had no feeling of change though they had practised PMR. The former category included the subcategories of experiences in which patients felt difficulty with PMR, experiences in which patients felt uncomfortable symptoms after PMR, and experiences in which patients did not feel it was good timing to learn PMR. CONCLUSION: It is easy for disease to progress in patients with cancer and those with poor strength, so it is necessary to match the functional status of individual patients. It also is important to support patients in practising PMR whenever they have the desire, by respecting their will. PMID- 20021582 TI - Personality traits affect individual interests in day service activities. AB - AIM: To assess the association of user interest in various activities with personality traits for the provision of activity programs at adult day centers. METHODS: A self-reported questionnaire survey of service users was conducted at 25 day centers in Tokyo; 133 men and 344 women, with a mean age of 81.6 +/- 7.9 years, responded. The questionnaire examined their demographics, lifestyles, requested activities, purpose of activity participation, attitudes toward participation, and personality traits by using the BASIC-3 Personality Inventory (PI)-short version. RESULTS: The participants with higher sociability and novelty seeking scores on the BASIC-3 PI-short version requested significantly more activities than those with lower scores. Music appreciation and singing were significantly associated with both sociability and novelty-seeking facets. The sociability facets were significantly associated with activities, such as walking and interaction with children, while the novelty-seeking facets were significantly associated with contact with animals and the Internet. There was no significant relationship between the neuroticism facets and any specific activity. Flower arrangement, handicrafts, fashion or make up, cooking, and singing were significantly more frequently requested by the women than by the men, while shogi or mahjong was requested significantly more frequently by the men than by the women. CONCLUSIONS: Sociability and novelty-seeking facets, sex, and age were significantly associated with an interest in particular activities. We recommend that adult day centers create a system under which nurses and formal caregivers fully assess the elderly in order to provide activity programs based on user needs. PMID- 20021584 TI - Systematic reviews in medicine. PMID- 20021585 TI - Tensions and paradoxes in electronic patient record research: a systematic literature review using the meta-narrative method. AB - CONTEXT: The extensive research literature on electronic patient records (EPRs) presents challenges to systematic reviewers because it covers multiple research traditions with different underlying philosophical assumptions and methodological approaches. METHODS: Using the meta-narrative method and searching beyond the Medline-indexed literature, this review used "conflicting" findings to address higher-order questions about how researchers had differently conceptualized and studied the EPR and its implementation. FINDINGS: Twenty-four previous systematic reviews and ninety-four further primary studies were considered. Key tensions in the literature centered on (1) the EPR ("container" or "itinerary"); (2) the EPR user ("information-processer" or "member of socio-technical network"); (3) organizational context ("the setting within which the EPR is implemented" or "the EPR-in-use"); (4) clinical work ("decision making" or "situated practice"); (5) the process of change ("the logic of determinism" or "the logic of opposition"); (6) implementation success ("objectively defined" or "socially negotiated"); and (7) complexity and scale ("the bigger the better" or "small is beautiful"). CONCLUSIONS: The findings suggest that EPR use will always require human input to recontextualize knowledge; that even though secondary work (audit, research, billing) may be made more efficient by the EPR, primary clinical work may be made less efficient; that paper may offer a unique degree of ecological flexibility; and that smaller EPR systems may sometimes be more efficient and effective than larger ones. We suggest an agenda for further research. PMID- 20021586 TI - Expensive cancer drugs: a comparison between the United States and the United Kingdom. AB - CONTEXT: This article compares the United Kingdom's and the United States' experiences with expensive cancer drugs to illustrate the challenges posed by new, extremely costly, medical technologies. METHODS: This article describes British and American coverage, access, and cost-sharing policies with regard to expensive cancer drugs and then compares the costs of eleven such drugs to British patients, American Medicare beneficiaries, and American patients purchasing the drugs in the retail market. Three questions posed by these comparisons are then examined: First, which system is fairer? In which system are cancer patients better off? Assuming that no system can sustainably provide to everyone at least some expensive cancer drugs for some clinical indications, what challenges does each system face in making these difficult determinations? FINDINGS: In both the British and American health care systems, not all patients who might benefit from or desire access to expensive cancer drugs have access to them. The popular characterization of the United States, where all cancer drugs are available for all to access as and when needed, and that of the British NHS, where top-down population rationing poses insurmountable obstacles to British patients' access, are far from the reality in both countries. CONCLUSIONS: Key elements of the British system are fairer than the American system, and the British system is better structured to deal with difficult decisions about expensive end-of-life cancer drugs. Both systems face common ethical, financial, organizational, and priority-setting challenges in making these decisions. PMID- 20021587 TI - Consumer-oriented health care reform strategies: a review of the evidence on managed competition and consumer-directed health insurance. AB - CONTEXT: For many years, leading health care reform proposals have been based on market-oriented strategies. In the 1990s, a number of reform proposals were built around the concept of "managed competition," but more recently, "consumer directed health care" models have received attention. Although price-conscious consumer demand plays a critical role in both the managed competition and consumer-directed health care models, the two strategies are based on different visions of the health care marketplace and the best way to use market forces to achieve greater systemwide efficiencies. METHODS: This article reviews the research literature that tests the main hypotheses concerning the two policy strategies. FINDINGS: Numerous studies provide consistent evidence that consumers' health plan choices are sensitive to out-of-pocket premiums. The elasticity of demand appears to vary with consumers' health risk, with younger, healthier individuals being more price sensitive. This heterogeneity increases the potential for adverse selection. Biased risk selection also is a concern when the menu of health plan options includes consumer-directed health plans. Several studies confirm that such plans tend to attract healthier enrollees. A smaller number of studies test the main hypothesis regarding consumer-directed health plans, which is that they result in lower medical spending than do more generous plans. These studies find little support for this claim. CONCLUSIONS: The experiences of employers that have adopted key elements of managed competition are generally consistent with the key hypotheses underlying that strategy. Research in this area, however, has focused on only a narrow range of questions. Because consumer-directed health care is such a recent phenomenon, research on this strategy is even more limited. Additional studies on both topics would be valuable. PMID- 20021588 TI - Aging in America in the twenty-first century: demographic forecasts from the MacArthur Foundation Research Network on an Aging Society. AB - CONTEXT: The aging of the baby boom generation, the extension of life, and progressive increases in disability-free life expectancy have generated a dramatic demographic transition in the United States. Official government forecasts may, however, have inadvertently underestimated life expectancy, which would have major policy implications, since small differences in forecasts of life expectancy produce very large differences in the number of people surviving to an older age. This article presents a new set of population and life expectancy forecasts for the United States, focusing on transitions that will take place by midcentury. METHODS: Forecasts were made with a cohort-components methodology, based on the premise that the risk of death will be influenced in the coming decades by accelerated advances in biomedical technology that either delay the onset and age progression of major fatal diseases or that slow the aging process itself. FINDINGS: Results indicate that the current forecasts of the U.S. Social Security Administration and U.S. Census Bureau may underestimate the rise in life expectancy at birth for men and women combined, by 2050, from 3.1 to 7.9 years. CONCLUSIONS: The cumulative outlays for Medicare and Social Security could be higher by $3.2 to $8.3 trillion relative to current government forecasts. This article discusses the implications of these results regarding the benefits and costs of an aging society and the prospect that health disparities could attenuate some of these changes. PMID- 20021589 TI - Translating medical effectiveness research into policy: lessons from the California Health Benefits Review Program. AB - CONTEXT: Legislatures and executive branch agencies in the United States and other nations are increasingly using reviews of the medical literature to inform health policy decisions. To clarify these efforts to give policymakers evidence of medical effectiveness, this article discusses the California Health Benefits Review Program (CHBRP). This program, based at the University of California, analyzes the medical effectiveness of health insurance benefit mandate bills for the California legislature, as well as their impact on cost and public health. METHODS: This article is based on the authors' experience reviewing benefit mandate bills for CHBRP and findings from evaluations of the program. General observations are illustrated with examples from CHBRP's reports. Information about efforts to incorporate evidence into health policymaking in other states and nations was obtained through a review of published literature. FINDINGS: CHBRP produces reports that California legislators, legislative staff, and other major stakeholders value and use routinely in deliberations about benefit mandate bills. Where available, the program relies on previously published meta-analyses and systematic reviews to streamline the review of the medical literature. Faculty and staff responsible for the medical effectiveness sections of CHBRP's reports have learned four major lessons over the course of the program's six-year history: the need to (1) recognize the limitations of the medical literature, (2) anticipate the need to inform legislators about the complexity of evidence, (3) have realistic expectations regarding the impact of medical effectiveness reviews, and (4) understand the consequences of the reactive nature of mandated benefit reviews. CONCLUSIONS: CHBRP has demonstrated that it is possible to produce useful reviews of the medical literature within the tight time constraints of the legislative process. The program's reports have provided state legislators with independent analyses that allow them to move beyond sifting through conflicting information from proponents and opponents to consider difficult policy choices and their implications. PMID- 20021590 TI - Research and advice giving: a functional view of evidence-informed policy advice in a Canadian Ministry of Health. AB - CONTEXT: As evidence-based medicine grows in influence and scope, its applicability to health policy prompts two questions: Can the principles and, more specifically, the tools used to bring research into the clinical world apply to civil servants offering advice to politicians? If not, what approach should the evidence-oriented health policy organization take to improve the use of research? METHODS: This article reviews evidence-based medicine and models of research use in policy. Then it reports the results of interviews with civil servants in the Ontario Ministry of Health, which recently adopted a stewardship rather than an operational role, incorporating many evidence-oriented strategies. The interviews focused on functional roles for research-based evidence in policy advice. FINDINGS: The clinical context and tools for evidence-based medicine can rarely be generalized to policy. Most current models of research use offer lessons to researchers wishing to apply their work to policy but little help for civil servants wishing to become more evidence oriented. The interviews revealed functional roles for research in setting agendas (noting upcoming issues and screening interest groups' claims), developing new policies (reducing uncertainty, helping speak truth to power, and preventing repetition and duplication), and monitoring or modifying existing policies (continuously improving programs and creating a culture of inquiry). Each area requires different tools to help filter the push of evidence from researchers and set agendas, to facilitate the urgent pull on relevant research by civil servants developing new policy, and to support ongoing linkage and exchange between civil servants and researchers for monitoring and modifying existing policy. CONCLUSIONS: A functional framework for evidence-informed policy advice is useful for distinguishing the activity from evidence-based medicine and "auditing" the balance of efforts across the different functional roles of research in policy. PMID- 20021592 TI - Phenology as a tool to link ecology and sustainable decision making in a dynamic environment. Symposium 14, 94th Ecological Society of America Meeting, Albuquerque, New Mexico, USA, August 2009. PMID- 20021591 TI - Weeds of agricultural importance: bridging the gap between evolutionary ecology and crop and weed science. PMID- 20021593 TI - From genes to ecosystems: an emerging synthesis of eco-evolutionary dynamics. Symposium 7, 94th Ecological Society of America Meeting, Albuquerque, New Mexico, USA, August 2009. PMID- 20021595 TI - President's message. AAPM expands the reach of Pain Medicine to address the public health problem of chronic pain. PMID- 20021596 TI - The effect of perceived racial discrimination on bodily pain among older African American men. AB - OBJECTIVES: We examined the extent to which experiences of racial discrimination are associated with bodily pain reported by African American men. METHODS: The study sample consisted of 393 African American male veterans who responded to a national survey of patients aged 50-75 who received care from the Veterans Health Administration (VHA). Veterans were surveyed by mail, with a telephone follow-up. The response rate for African Americans in the sample was 60.5%. Pain (assessed using the bodily pain subscale of the 36-item short-form health survey), experiences of discrimination, employment, education, and income were obtained through the survey. Age, race, and mental health comorbidities were obtained from VA administrative data. Multiple regression analysis adjusting for item non response (via imputation) and unit non-response (via propensity scores and weighting) was used to assess the association between racial discrimination and likelihood of experiencing moderate or severe pain over the past 4 weeks. RESULTS: Experiences of racial discrimination were associated with greater bodily pain (beta = -0.25, P < 0.0001), even after controlling for socioeconomic and health-related characteristics. CONCLUSION: Perceived racial discrimination was associated with greater pain among a sample of older African American male patients in the VA. Additional research is needed to replicate this finding among other populations of African Americans. PMID- 20021597 TI - Ultrasound-guided permanent implantation of peripheral nerve stimulation (PNS) system for neuropathic pain of the extremities: original cases and outcomes. AB - OBJECTIVE: Our objective is to describe our experience applying a minimally invasive, ultrasound-assisted technique for peripheral nerve stimulation (PNS) trial and permanent implantation in patients with neuropathic extremity pain. DESIGN: Retrospective case series and review of the literature. Setting. Tertiary referral center and academic medical center. PATIENTS: Patients with upper or lower extremity neuropathic pain resistant to other therapies who responded to an ultrasound-guided peripheral nerve block at a proximal location. Interventions. Ultrasound-assisted through-the-needle placement of percutaneous neurostimulation electrodes on target major peripheral nerves for fully percutaneous trial, staged trial or permanent implantation of PNS system. A PNS trial period of 3-7 days was used. OUTCOME MEASURES: Pain relief at last follow-up, complications, therapeutic limitations due to technique as applied. Results. Six of eight (75%) patients and 7/9 (78%) peripheral nerves had a successful trial and underwent permanent PNS system implantation using a minimally invasive, ultrasound-assisted technique from November 2007 to December 2008. All but one patient with an implanted PNS system had > or =50% pain relief at last follow-up and 3/7 (43%) permanent systems were associated with > or =80% relief. Loss of paresthesia required revision to dual-lead systems in upper extremity radial nerve PNS. Infection led to explant in one case. CONCLUSIONS: In a small series of patients, a minimally invasive, ultrasound-assisted technique for PNS trial, and permanent PNS implantation proved feasible. Patients without adequate analgesia during neurostimulation trial avoided surgical incision and those undergoing permanent implantation were not subjected to the potential morbidity associated with nerve dissection. PMID- 20021598 TI - Long-term adjustment to work-related low back pain: associations with socio demographics, claim processes, and post-settlement adjustment. AB - OBJECTIVE: Predict long-term adjustment (pain intensity, pain-related catastrophizing, and pain-related disability) from socio-demographic, claim process, and post-settlement adjustment variables in a cohort of 374 Workers' Compensation low back claimants. METHODS: Age- and gender-matched subsamples of African Americans and Caucasians were randomly selected for long-term follow-up (6 years post-settlement) from a larger, existing cohort of Workers' Compensation low back claimants in Missouri. Computer-assisted telephone interviews were used to assess pain, catastrophizing, and disability. Path analysis and logistic regression analysis were used to predict long-term adjustment from socio demographic variables (race, gender, age, and socioeconomic status), Workers' Compensation claim process variables (surgery, diagnosis, claim duration, treatment costs, settlement awards, and disability rating), and adjustment at baseline. RESULTS: Poorer long-term adjustment (higher levels of pain, catastrophizing, and pain-related disability) was significantly predicted by relatively poorer adjustment at baseline, lower socioeconomic status, and African American race. African American race associations were also mediated through lower socioeconomic status. Higher levels of occupational disability, as measured by long-term rates of unemployment and social security disability, were also predicted by African American race (in addition to age and claim process factors). CONCLUSION: Long-term adjustment to low back pain in this cohort of Workers' Compensation claimants was stable, relative to short-term adjustment soon after settlement. Long-term adjustment was worse for people of lower socioeconomic status, particularly for economically disadvantaged African Americans, suggesting the possibility of race- and class-based disparities in the Workers' Compensation system. PMID- 20021600 TI - Chronic pain in German general practice. AB - INTRODUCTION: This study evaluates the prevalence of chronic pain, intensity of pain, activity limitation, and pain-related diagnoses in German general practices. METHODS: In 40 general practices, up to 50 consecutive patients presenting to general practitioners (GP) for routine medical consultation were questioned, and those reporting pain that lasted for more than 3 months received a questionnaire referring to intensity of pain and activity limitations. GPs received a questionnaire asking about the duration of treatment and diagnoses. RESULTS: Three hundred forty-six out of 1,860 questioned patients suffered from pain that lasted for more than 3 months (a point prevalence of 18.4% [95% confidence interval 16.7-20.3]). The average degree of pain equaled 5 out of 10 points on a numerical grading scale (NRS); the average degree of activity limitation was 4.8 out of 10. In most cases, the pain was related to musculoskeletal degenerative diseases. CONCLUSION: Chronic pain patients constitute a considerable share of workload in general practice. PMID- 20021599 TI - A mind-body program for older adults with chronic low back pain: results of a pilot study. AB - OBJECTIVES: Determine the impact of an 8-week mindfulness meditation program on disability, psychological function, and pain severity in community-dwelling older adults with chronic low back pain, and to test the education control program for feasibility. DESIGN: Randomized controlled trial. Participants. Forty community dwelling older adults with moderate low back pain or greater for at least the previous 3 months. Intervention. Participants were randomized to an 8-week meditation program or an 8-week education control program. OUTCOME MEASURES: Disability, psychological function, and pain severity were assessed. The same measures were obtained for both groups at baseline, at the end of the program, and 4 months after program completion. RESULTS: Sixteen participants (80%) completed the meditation program and 19 (95%) completed the education program. Both the meditation and control group improved on measures of disability, pain, and psychological function, both at program completion and 4-month follow-up. The differences between the two groups did not reach statistical significance. The meditation group practiced mindfulness meditation a mean of 5 days/week (range 1 7) and mean of 31 minutes/session (range 22-48). At 4 months follow-up 14/16 (88%) participants continued to meditate. CONCLUSION: Both the intervention group and the education control group improved on outcome measures suggesting both programs had a beneficial effect. Participants continued to meditate on 4-month follow-up. The control program was feasible but not inert. Piloting the control program in mind-body research can inform the design of larger clinical trials. PMID- 20021601 TI - A comparison of common screening methods for predicting aberrant drug-related behavior among patients receiving opioids for chronic pain management. AB - OBJECTIVE: The ability to predict risk for violating opioid medication policies, known as aberrant drug-related behavior, is critical for providing optimal treatment. Many pain management centers measure risk using one of several partially validated measures: the Screener and Opioid Assessment for Patients with Pain (SOAPP), the Diagnosis, Intractability, Risk, and Efficacy inventory (DIRE), and/or the Opioid Risk Tool (ORT). However, little is known about how these measures compare with each other in predicting aberrant drug-related behavior and discontinuance of opioid pain medications. The current study aimed to address this research question. PATIENTS: Participants were 48 patients who attended a pain management center in Tennessee but were later discontinued from opioids for aberrant drug-related behavior. Patients referred for opioid medication for pain management participated in a semi-structured clinical interview with the staff psychologist and completed the aforementioned measures. Patients generally returned to the pain clinic on a monthly basis for medication management. Results. Analyses compared the sensitivity of each self-report measure and the clinical interview in predicting discontinuance for aberrant drug related behavior. RESULTS: showed the highest sensitivity for the clinical interview (0.77) and the SOAPP (0.72), followed by the ORT (0.45) and the DIRE (0.17). Combining the clinical interview with the SOAPP increased sensitivity to 0.90. CONCLUSIONS: Among patients who were discontinued from opioids for aberrant drug-related behaviors, the clinical interview and the SOAPP were most effective at predicting risk at baseline. Implications for future research and clinical practice are discussed. PMID- 20021602 TI - Commentary: "electroconvulsive stimulation (ECS) increases the expression of neuropeptide Y (NPY) in rat brains in a model of neuropathic pain: a quantitative real-time polymerase chain reaction (RT-PCR) study," by Okabe and colleagues. PMID- 20021603 TI - Restless legs syndrome and painful neuropathy-retrospective study. A role for nociceptive deafferentation? AB - OBJECTIVES: Restless legs syndrome (RLS) occurs in polyneuropathy with small fiber involvement, possibly as a peculiar form of neuropathic pain; however, the relationship between pain and RLS has been poorly investigated in polyneuropathy. DESIGN, SETTING, AND PATIENTS: We evaluated retrospectively the occurrence of RLS in 102 consecutive patients with polyneuropathy manifesting with neuropathic pain or dysesthesia, referred to the Neuromuscular Center, using the National Institutes of Health criteria for RLS. The patients were classified in subgroups characterized respectively by allodynia (hyperphenomena), with reported unpleasant sensations evoked by tactile stimuli, and hypoalgesia (hypophenomena), with absent pain sensation to pinprick, according to putative mechanisms of pain. RESULTS: RLS was present in 41/102 patients (40.2%). It was significantly more frequent in the "hypoalgesia" (23/37) than in the "allodynia" subgroup (9/31; P = 0.008) and in the not classifiable cases (9/34; P = 0.004). CONCLUSIONS: RLS is frequent in painful polyneuropathy and is significantly associated with decreased small fiber input, thus nociceptive deafferentation may represent a factor interacting with RLS "generators," possibly at spinal level. We suggest that overactivity of the spinal structures implicated in RLS may be triggered by nociceptive deafferentation in a subgroup of patients with painful polyneuropathy. Our findings, prompting a mechanistic characterization of RLS associated with painful polyneuropathy, have to be confirmed in a prospective study. PMID- 20021604 TI - Re: Bussing A, Michalsen A, Balzat H-J, Grunther RA, Ostermann T, Neugebauer EAM, and Mathiessen PF. Are spirituality and religiosity resources for patients with chronic pain conditions? Pain Medicine 2009; 10: 327-339. PMID- 20021605 TI - Are we treating pain or patients suffering from pain? PMID- 20021607 TI - Age-related EBV-associated B-cell lymphoproliferative disorders: diagnostic approach to a newly recognized clinicopathological entity. AB - EBV is prevalent among healthy individuals, and is implicated in numerous reactive and neoplastic processes in the immune system. The authors originally identified a series of senile or age-related EBV-associated B-cell lymphoproliferative disorders (LPD) bearing a resemblance to immunodeficiency associated ones, which may be associated with immune senescence in the elderly and which are now incorporated into the 2008 World Health Organization lymphoma classification as EBV-positive diffuse large B-cell lymphoma (DLBCL) of the elderly. This newly described disease is pathologically characterized by a proliferation of atypical large B cells including Reed-Sternberg-like cells with reactive components, which pose a diagnostic problem for pathologists. Clinically, this disease may present with lymphadenopathy, and is often extranodal, frequently involving the skin, gastrointestinal tract, or lung. Onset is usually after the age of 50; the median patient age is 70-79 years, and incidence continues to increase with age, providing additional support to the nosological term of EBV+ DLBCL of the elderly. These patients have a worse prognosis than those with EBV-negative DLBCL or EBV+ classical Hodgkin lymphoma (CHL). The aim of the present review was to summarize the clinicopathological profile of age-related EBV+ LPD and EBV+ Hodgkin lymphoma to facilitate diagnostic approach. PMID- 20021608 TI - Sclerosing angiomatoid nodular transformation of the spleen (SANT): clinicopathological study of 10 cases with or without abdominal disseminated calcifying fibrous tumors, and the presence of a significant number of IgG4+ plasma cells. AB - Sclerosing angiomatoid nodular transformation (SANT) is a peculiar splenic vascular lesion that is characterized by marked stromal sclerosis and the presence of plasma cells, which shares histopathological features associated with IgG4-related sclerosing disease. The cinicopathological features of 10 cases of SANT were reviewed and immunohistochemistry with IgG4 and IgG antibodies was performed. Nine other various splenic lesions served as controls. Five cases of SANT were found incidentally. Three cases of SANT had multiple tumors and five had associated abdominal disseminated calcifying fibrous tumors (CFT). IgG4+ plasma cells were found in all of the cases of SANT and in calcifying fibrous tumors. The densities of IgG4+ and IgG+ cells and the IgG4/IgG ratios were significantly higher in SANT than in control spleens (P= 0.001, 0.006, and 0.028, respectively). Serum IgG4 concentration was elevated in one case. In conclusion, SANT can occur either as a solitary or as multiple tumors. Frequent association with abdominal disseminated CFT simulating carcinomatosis was observed. A statistically significant number of IgG4+ plasma cells was found in all of the cases of SANT and the associated CFT. Whether they are related to the IgG4 related sclerosing disease or not is debatable and warrants further investigation. PMID- 20021609 TI - Comparison of tissue microarray and full section in immunohistochemistry of gastrointestinal stromal tumors. AB - The aim of the present study was to compare the efficiency of tissue microarray (TMA) results using immunohistochemistry markers applied to a variety of core sizes and full sections of gastrointestinal stromal tumors (GIST) using performance measures such as core loss rate and concordance rate. Six primary antibody markers (c-Kit, CD34, smooth muscle actin (SMA), S-100, p53, and Ki-67) were used with the TMA technique to analyze 0.6 mm, 2 mm, and 3 mm punch cores of GIST samples from 67 patients. No statistical association was found between core size and loss rate (P= 0.512). TMA results for the 0.6 mm, 2 mm, and 3 mm core showed that all core sizes could statistically significantly reflect full sections with regard to c-Kit, SMA, and S-100 antibodies, but that the 3 mm core section was the most representative except for CD34. With regard to p53 and Ki-67 staining, the 0.6 mm core section was not representative, but the 2 mm and 3 mm core sections could statistically significantly represent full section results. Among them, the 3 mm core section was more accurate than the 2 mm core section. Use of a single 3 mm core size in TMA is suitable for evaluating large numbers of protein and nuclear stains with regard to immunohistochemistry for GIST. PMID- 20021610 TI - Area of residual tumor beyond the muscular layer is a useful predictor of outcome in rectal cancer patients who receive preoperative chemoradiotherapy. AB - The purpose of the present study was to determine whether the amount and the location of residual tumor are associated with outcome in surgically treated rectal cancer patients who receive preoperative chemoradiation therapy. Forty three rectal cancer patients who underwent sphincter-saving operations after preoperative chemoradiation therapy were enrolled in the study. The total area of residual tumors was measured using morphometry software, and then the area of the residual tumors located within and beyond the muscular layer was also determined. Associations between clinicopathological features were evaluated. The results showed that the total area of residual tumor and area of residual tumor within the muscular layer were associated with TNM stage, tumor regression, and microscopy features, but not with patient disease-free survival. The area of the residual tumor located beyond the muscular layer was significantly associated with pathological ypT, ypN stage, tumor downstaging, perineural invasion, and the depth of tumor invasion beyond the muscular layer (P < 0.05). Further, large residual tumor area beyond the muscular layer was associated with shorter disease free survival (P < 0.05). Morphometry of residual tumor area beyond the muscular layer is a new pathological prognostic factor for rectal cancer patients receiving preoperative chemoradiation therapy. PMID- 20021611 TI - Expression of ERCC1 and class IIIbeta tubulin for predicting effect of carboplatin/paclitaxel in patients with advanced inoperable non-small cell lung cancer. AB - It was recently reported that expression of excision repair cross-complementation group 1 (ERCC1), a DNA repair protein, predicts sensitivity to platinum-based chemotherapy drugs. Microtubule inhibitors such as paclitaxel demonstrate anticancer effects by inhibiting spindle fibers during mitosis; and class IIIbeta tubulin (IIIbeta tubulin), a microtubule component, is thought to be resistant to microtubule inhibitors. The purpose of the present study was to examine the correlation between prognosis and expression of these proteins using biopsy tissues obtained from 40 patients with advanced inoperable non-small cell lung cancer who had been treated with carboplatin plus Taxol. On immunostaining 27 patients (68%) were positive for ERCC1 and 22 (55%) were positive for IIIbeta tubulin. The prognosis of the ERCC1-negative group was significantly better than that for the ERCC1-positive group (P= 0.014). As for IIIbeta tubulin, the prognosis for the negative group was also significantly better than that for the positive group (P= 0.025). Multivariate analysis showed that the expression of ERCC1 was an independent predictor of prognosis (hazard ratio: 3.485; 95% confidence interval: 1.123-10.818, P= 0.031). It was concluded that determination of the expression of these proteins is useful to predict the effects of platinum based anticancer drugs. PMID- 20021612 TI - Plasmablastic lymphoma of the retroperitoneum in an HIV-negative patient. AB - Herein is reported a case of plasmablastic lymphoma (PBL) of the retroperitoneum in an HIV-negative patient. This is the first reported case of PBL at this location and of PBL from Japan in the English-language literature. A 76-year-old Japanese man was admitted to hospital with a chief complaint of right inguinal lymph node swelling. Lymph node biopsy indicated large tumor cells with both diffuse and cohesive growth patterns, and conspicuous tumor cell proliferation in lymph node sinuses. The initial pathological diagnosis was metastatic carcinoma. The patient died approximately 1 month after admission, and autopsy showed that the main lesion was a very large retroperitoneal mass. On histology diffusely proliferated plasmablast-like or immunoblast-like tumor cells were identified, which were positive on immunohistochemistry for CD138 and negative for B-cell and epithelial markers. Approximately 90% of the tumor cells were positive for Ki-67. Tumor cells were diffusely positive for EBV-encoded small RNA on in situ hybridization. The autopsy findings suggested a diagnosis of PBL. Accordingly, PBL should be considered as a differential diagnosis when lymph node biopsy findings resemble those of the present patient. PMID- 20021613 TI - Pleural malignant mesothelioma with invasive micropapillary component and its association with pulmonary metastasis. AB - The micropapillary pattern (characterized by papillary structure with tufts lacking a central fibrovascular core) is a predictor of aggressive carcinoma. The purpose of the present study was to review 34 pleural malignant mesotheliomas (21 epithelioid, five sarcomatoid, seven biphasic and one lymphohistiocytoid), with special reference to the presence of invasive micropapillary component. Two invasive micropapillary pattern-positive tumors were identified. The invasive micropapillary pattern was seen to have a focal distribution in 15-20% of the tumor tissues. The majority of the invasive micropapillary clusters expressed MUC1 along the outer cell surface. Analysis of pleural malignant mesotheliomas with epithelioid features and with or without invasive micropapillary pattern (21 epithelioid and seven biphasic subtypes) indicated pulmonary micrometastases in only the invasive micropapillary-positive tumors (P < 0.015), and the spread was probably via the lymphatics. Lymphatic involvement (confirmed on immunohistochemistry with D2-40 antibody) and lymph node metastasis were found in both of the invasive micropapillary-positive tumor patients, whereas they were noted in only one of 10 (10%, P < 0.046) and three of nine (30%) invasive micropapillary-negative patients. To the authors' knowledge this is the first study to indicate the presence of invasive micropapillary component in pleural malignant mesothelioma. This component can predict more aggressive lymphatic spread, similar to that of carcinomas in other organs with micropapillary pattern. PMID- 20021614 TI - Collision tumor of the stomach: gastric adenocarcinoma intermixed with gastrointestinal stromal tumor. AB - The synchronous occurrence of tumors of different histological types in the stomach is uncommon. This rare association is usually detected incidentally at gastroscopy, surgery or histopathology. It is still not known whether such association represents an incidental coexistence or indicates a similar pathogenesis in the simultaneous development of tumors of different histological types in the same organ. A 73-year-old woman complained of discomfort after eating a Chinese dumpling, and had tarry stools three times. Gastroscopy showed a tumor at the juncture of the gastric fundus and corpus. The tumor was composed of gastric adenocarcinoma and gastrointestinal stromal tumor, in which the two components were intermixed with each other. To the authors' knowledge there are few reports of this extremely rare collision tumor association. PMID- 20021615 TI - Clinicopathological features of syphilitic gastritis in Korean patients. AB - Syphilitic gastritis is difficult to diagnose without a history of syphilis, because endoscopic and microscopic findings can simulate gastric cancer or lymphoma. Herein are reported two cases of syphilitic gastritis and analysis of the clinicopathological characteristics of 11 cases in Korean subjects, including nine previously reported ones. The two present patients were 25- and 32-year-old men with epigastric pain. The gastroduodenoscopy features were similar: multiple erosive or ulcerative lesions in whole gastric mucosa. Microscopically, gastric mucosa contained severe chronic active gastritis involving submucosa with crypt abscesses, venulitis and granulomas, suggesting unusual infectious gastritis. Numerous spirochetes were identified on silver staining, and serological tests for syphilis were reactive in both patients. They were treated with benzathin penicillin, and follow-up gastric biopsy specimens indicated non-specific chronic gastritis without evidence of syphilis. Of the 11 Korean patients with syphilitic gastritis, six were female and five were male (mean age, 31.7 years). The most common symptom was epigastric pain and all patients had diffuse erosive, ulcerative or infiltrative lesions on gastroduodenoscopy. Syphilitic gastritis should be considered in young patients with epigastric pain who have diffuse gastric involvement such as erosion or shallow ulcer on endoscopy and unusually extensive chronic active gastritis on microscopy, even if the history of syphilis is not clear. PMID- 20021616 TI - Unique vascular tumor primary arising in the liver and exhibiting histopathological features consistent with so-called polymorphous hemangioendothelioma. AB - Reported herein is an unusual vascular tumor primary arising in the liver and exhibiting unique histopathological features. A 47-year-old woman underwent left hepatectomy because of a large hepatic mass. On histology the tumor had a composite pattern, consisting of angiomatous, retiform and solid areas, formed by oval to cuboidal to spindle cells, that expressed only endothelial markers (CD31 and factor VIII-related antigen). These findings led to the diagnosis of a low grade vascular neoplasm with morphological features consistent with so-called polymorphous hemangioendothelioma. The tumor was completely resected. At 24 month follow up the patient was alive, without evidence of disease. Polymorphous hemangioendothelioma is a rare vascular neoplasm, with borderline malignant potential, which usually occurs in lymph nodes and, rarely, at extranodal sites. Its classification as an entity has been questioned recently. The unusual morphological features of the present case, which do not fit neatly with any other recognized hemangioendothelioma subtype, indicate that the family of vascular tumors is broader than currently accepted. In addition the present case widens the spectrum of primary vascular tumors arising in the liver. PMID- 20021617 TI - Quantitative assessment of gene methylation in neoplastic and non-neoplastic gastric epithelia using methylation-specific DNA microarray. AB - A fiber-type DNA microarray was used to calculate methylation rates (MR) of four tumor suppressor genes, lysyl oxidase (LOX), p16, RUNX3, and tazarotene-induced gene 1 (TIG1). MR were calculated in 26 primary gastric cancers and corresponding non-neoplastic gastric epithelia, and the results were compared to those of conventional methylation-specific polymerase chain reaction (MSP). MR ranged from 0.1% to 69.1% (mean, 18.3%) for LOX, 0.5-74.1% (mean, 15.7%) for p16, 0.2-76.5% (mean, 22.7%) for RUNX3, and 0.6-41.2% (mean, 5.8%) for TIG1 in primary gastric cancers, and from 0.1% to 25.8% (mean, 8.7%) for LOX, 1.0- 23.2% (mean, 10.3%) for p16, 0.7-25.1% (mean, 5.5%) for RUNX3, and 1.8-27.6% (mean, 11.4%) for TIG1 in corresponding non-neoplastic gastric epithelia. Although MR varied significantly across different samples for both neoplastic and non-neoplastic gastric epithelia, high-level methylation (MR >40%) was cancer specific and was observed in 19.2%, 19.2%, 30.8%, and 3.8% of primary gastric cancers for LOX, p16, RUNX3, and TIG1, respectively. All samples with high-level methylation, as well as some samples with low MR (particularly <10%) were judged to be methylation positive on conventional MSP. Quantitative analysis of gene methylation using methylation-specific DNA microarray is a promising method for cancer diagnosis. PMID- 20021619 TI - Cardiovascular disease in special populations III: stroke. PMID- 20021620 TI - Coronary stents and noncardiac surgery: current clinical challenges and conundrums. AB - This article discusses how perioperative physicians are currently faced with unique challenges when providing care for surgical patients who have undergone recent percutaneous transluminal coronary angioplasty with drug-eluting stent (DES) placement. Despite adhering to the currently recommended antiplatelet regimens, these patients may still be at risk for sustaining a myocardial infarction secondary to stent thrombosis during the perioperative period. Given the high morbidity and mortality rates associated with perioperative myocardial infarction, it is important that the perioperative care of surgical patients with previous DES placement should be discussed, evaluated, and clarified by all practitioners who may be involved in their care. PMID- 20021621 TI - Incidence of myocardial infarction or stroke or death at 47-month follow-up in patients with diabetes and a predicted exercise capacity 85% during an exercise treadmill sestamibi stress test. AB - A treadmill exercise sestamibi stress test (TESST) was performed in 609 consecutive diabetic persons with a mean age of 70 years and no history of coronary artery disease (CAD) who were referred for a TESST because of chest pain or dyspnea. Of 609 patients, 301 (49%) had a predicted exercise capacity 85% (group B). Group A patients had a higher prevalence of myocardial ischemia (43% vs 30%, P=.0005), 2- or 3-vessel obstructive CAD (38% vs 18%, P=.001), myocardial infarction (17% vs 9%, P=.004), death (10% vs 4%, P=.008), and myocardial infarction or stroke or death at 47-month follow-up (21% vs 12%, P=.001). Stepwise Cox regression analysis showed that the only significant independent predictor for the time to development of myocardial infarction or stroke or death was a predicted exercise capacity >85% (hazard ratio, 0.52; 95% confidence interval, 0.34-0.78; P=.002). Diabetic persons with a predicted exercise capacity >85% had a 48% lower chance of myocardial infarction, stroke, or death than those with a predicted exercise capacity C) and MDM2 SNP309 ( 410T>G), and p53 protein expression in breast tumors on survival. METHODS: We pooled data from four breast cancer cohorts within the Breast Cancer Association Consortium for which both TP53 R72P and MDM2 SNP309 were genotyped and follow-up was available (n = 3,749). Overall and breast cancer-specific survival analyses were performed using Kaplan-Meier analysis and multivariate Cox's proportional hazards regression models. RESULTS: Survival of patients did not differ by carriership of either germ-line variant, R72P (215G>C) or SNP309 (-410G>T) alone. Immunohistochemical p53 staining of the tumor was available for two cohorts (n = 1,109 patients). Survival was worse in patients with p53-positive tumors (n = 301) compared to patients with p53-negative tumors (n = 808); breast cancer specific survival: HR 1.6 (95% CI 1.2 to 2.1), P = 0.001. Within the patient group with p53-negative tumors, TP53 rare homozygous (CC) carriers had a worse survival than G-allele (GG/GC) carriers; actuarial breast cancer-specific survival 71% versus 80%, P = 0.07; HR 1.8 (1.1 to 3.1), P = 0.03. We also found a differential effect of combinations of the two germ-line variants on overall survival; homozygous carriers of the G-allele in MDM2 had worse survival only within the group of TP53 C-allele carriers; actuarial overall survival (GG versus TT/TG) 64% versus 75%, P = 0.001; HR (GG versus TT) 1.5 (1.1 to 2.0), P = 0.01. We found no evidence for a differential effect of MDM2 SNP309 by p53 protein expression on survival. CONCLUSIONS: The TP53 R72P variant may be an independent predictor for survival of patients with p53-negative tumors. The combined effect of TP53 R72P and MDM2 SNP309 on survival is in line with our a priori biologically-supported hypothesis, that is, the role of enhanced DNA repair function of the TP53 Pro-variant, combined with increased expression of the Mdm2 protein, and thus overall attenuation of the p53 pathway in the tumor cells. PMID- 20021640 TI - Temporal and spatial stability of Anopheles gambiae larval habitat distribution in Western Kenya highlands. AB - BACKGROUND: Localized mosquito larval habitat management and the use of larvicides have been proposed as important control tools in integrated malaria vector management programs. In order to optimize the utility of these tools, detailed knowledge of the spatial distribution patterns of mosquito larval habitats is crucial. However, the spatial and temporal changes of habitat distribution patterns under different climatic conditions are rarely quantified and their implications to larval control are unknown. RESULTS: Using larval habitat data collected in western Kenya highlands during both dry and rainy seasons of 2003-2005, this study analyzed the seasonal and inter-annual changes in the spatial patterns in mosquito larval habitat distributions. We found that the spatial patterns of larval habitats had significant temporal variability both seasonally and inter-annually. CONCLUSIONS: The pattern of larval habitats is extremely important to the epidemiology of malaria because it results in spatial heterogeneity in the adult mosquito population and, subsequently, the spatial distribution of clinical malaria cases. Results from this study suggest that larval habitat management activities need to consider the dynamic nature of malaria vector habitats. PMID- 20021641 TI - Effect of provider-initiated testing and counselling and integration of ART services on access to HIV diagnosis and treatment for children in Lilongwe, Malawi: a pre- post comparison. AB - BACKGROUND: The HIV prevalence in Malawi is 12% and Kamuzu Central Hospital (KCH), in the capital Lilongwe, is the main provider of adult and paediatric HIV services in the central region. The Lighthouse at KCH offers opt-in HIV testing and counselling (HTC) for adults and children. In June 2004, Lighthouse was the first clinic to provide free antiretroviral treatment (ART) in the public sector, but few children accessed the services. In response, provider-initiated HIV testing and counselling (PITC) and an ART clinic were introduced at the paediatric department at KCH in Quarter 4 (Q4) 2004. METHODS: We analysed prospectively collected, aggregated data of quarterly reports from Q1 2003 to Q4 2006 from HTC centre registers, ART registers and clinic registrations at the ART clinics of both Lighthouse and the paediatric department. By comparing data of both facilities before (Q1 2003 to Q3 2004), and after the introduction of the services at the paediatric department (Q4 2004 to Q4 2006), we assessed the effect of this intervention on the uptake of HIV services for children at KCH. RESULTS: Overall, 3971 children were tested for HIV, 2428 HIV-infected children were registered for care and 1218 started ART. Between the two periods, the median (IQR) number of children being tested, registered and starting ART per quarter rose from 101 (53-109) to 358 (318-440), 56 (50-82) to 226 (192-234) and 18 (8-23) to 139 (115-150), respectively. The median proportion of tested clients per quarter that were children rose from 3.8% (2.7-4.3) to 9.6% (8.8 to 10.0) (p = 0.0009) and the proportion of ART starters that were children rose from 6.9% (4.9-9.3) to 21.1% (19.2-24.2) (p = 0.0036). The proportion of registered children and adults starting ART each quarter increased similarly, from 26% to 53%, and 20% to 52%, respectively. CONCLUSIONS: Implementation of PITC and integration of ART services within the paediatric ward are likely to be the main reasons for improved access to HTC and ART for children at KCH, and can be recommended to other hospitals with paediatric inpatients in resource limited settings with high HIV prevalence. PMID- 20021642 TI - Genetic tools for the investigation of Roseobacter clade bacteria. AB - BACKGROUND: The Roseobacter clade represents one of the most abundant, metabolically versatile and ecologically important bacterial groups found in marine habitats. A detailed molecular investigation of the regulatory and metabolic networks of these organisms is currently limited for many strains by missing suitable genetic tools. RESULTS: Conjugation and electroporation methods for the efficient and stable genetic transformation of selected Roseobacter clade bacteria including Dinoroseobacter shibae, Oceanibulbus indolifex, Phaeobacter gallaeciensis, Phaeobacter inhibens, Roseobacter denitrificans and Roseobacter litoralis were tested. For this purpose an antibiotic resistance screening was performed and suitable genetic markers were selected. Based on these transformation protocols stably maintained plasmids were identified. A plasmid encoded oxygen-independent fluorescent system was established using the flavin mononucleotide-based fluorescent protein FbFP. Finally, a chromosomal gene knockout strategy was successfully employed for the inactivation of the anaerobic metabolism regulatory gene dnr from D. shibae DFL12T. CONCLUSION: A genetic toolbox for members of the Roseobacter clade was established. This provides a solid methodical basis for the detailed elucidation of gene regulatory and metabolic networks underlying the ecological success of this group of marine bacteria. PMID- 20021643 TI - Pancreatic adenocarcinoma in a patient with situs inversus: a case report of this rare coincidence. AB - BACKGROUND: Situs inversus (SI) is a relatively rare occurrence in patients with pancreatic adenocarcinoma. Pancreatic resection in these patients has rarely been described. CT scan imaging is a principle modality for detecting pancreatic cancer and its use in SI patients is seldom reported. CASE PRESENTATION: We report a 48 year old woman with SI who, despite normal CT scan 8 months earlier, presented with obstructive jaundice and a pancreatic head mass requiring a pancreaticoduodenectomy. The surgical pathology report demonstrated pancreatic adenocarcinoma. CONCLUSION: SI is a rare condition with concurrent pancreatic cancer being even rarer. Despite the rarity, pancreaticoduodenectomy in these patients for resectable lesions is safe as long as special consideration to the anatomy is taken. Additionally, radiographic imaging has significantly improved detection of early pancreatic cancer; however, there continues to be a need for improved detection of small neoplasms. PMID- 20021644 TI - YsxC, an essential protein in Staphylococcus aureus crucial for ribosome assembly/stability. AB - BACKGROUND: Bacterial growth and division requires a core set of essential proteins, several of which are still of unknown function. They are also attractive targets for the development of new antibiotics. YsxC is a member of a family of GTPases highly conserved across eubacteria with a possible ribosome associated function. RESULTS: Here, we demonstrate by the creation of a conditional lethal mutant that ysxC is apparently essential for growth in S. aureus. To begin to elucidate YsxC function, a translational fusion of YsxC to the CBP-ProteinA tag in the staphylococcal chromosome was made, enabling Tandem Affinity Purification (TAP) of YsxC-interacting partners. These included the ribosomal proteins S2, S10 and L17, as well as the beta' subunit of the RNA polymerase. YsxC was then shown to copurify with ribosomes as an accessory protein specifically localizing to the 50 S subunit. YsxC depletion led to a decrease in the presence of mature ribosomes, indicating a role in ribosome assembly and/or stability in S. aureus. CONCLUSIONS: In this study we demonstrate that YsxC of S. aureus localizes to the ribosomes, is crucial for ribosomal stability and is apparently essential for the life of S. aureus. PMID- 20021645 TI - Suppression of MMP activity in bovine cartilage explants cultures has little if any effect on the release of aggrecanase-derived aggrecan fragments. AB - BACKGROUND: Progressive loss of articular cartilage is a central hallmark in many joint disease, however, the relative importance of individual proteolytic pathways leading to cartilage erosion is at present unknown. We therefore investigated the time-dependant release ex vivo of MMP- and aggrecanase-derived fragments of aggrecan and type II collagen into the supernatant of bovine cartilage explants cultures using neo-epitope specific immunoassays, and to associate the release of these fragments with the activity of proteolytic enzymes using inhibitors. FINDINGS: Bovine cartilage explants were cultured in the presence or absence of the catabolic cytokines oncostatin M (OSM) and tumor necrosis factor alpha (TNFalpha). In parallel, explants were co-cultured with protease inhibitors such as GM6001, TIMP1, TIMP2 and TIMP3. Fragments released into the supernatant were determined using a range of neo-epitope specific immunoassays; (1) sandwich (342)FFGVG-G2 ELISA, (2) competition NITEGE(373)ELISA (3) sandwich G1-NITEGE(373 )ELISA (4) competition (374)ARGSV ELISA, and (5) sandwich (374)ARGSV-G2 ELISA all detecting aggrecan fragments, and (6) sandwich CTX-II ELISA, detecting C-telopeptides of type II collagen. We found that (1) aggrecanase-derived aggrecan fragments are released in the early (day 2-7) and mid phase (day 9-14) into the supernatant from bovine explants cultures stimulated with catabolic cytokines, (2) the release of NITEGE(373 )neo-epitopes are delayed compared to the corresponding (374)ARGSV fragments, (3) the MMP inhibitor GM6001 did not reduce the release of aggrecanase-derived fragment, but induced a further delay in the release of these fragments, and finally (4) the MMP-derived aggrecan and type II collagen fragments were released in the late phase (day 16-21) only. CONCLUSION: Our data support the model, that aggrecanases and MMPs act independently in the processing of the aggrecan molecules, and furthermore that suppression of MMP-activity had little if any effect on the quantity of aggrecanase-derived fragments released from explants cultures. PMID- 20021646 TI - WebCARMA: a web application for the functional and taxonomic classification of unassembled metagenomic reads. AB - BACKGROUND: Metagenomics is a new field of research on natural microbial communities. High-throughput sequencing techniques like 454 or Solexa-Illumina promise new possibilities as they are able to produce huge amounts of data in much shorter time and with less efforts and costs than the traditional Sanger technique. But the data produced comes in even shorter reads (35-100 basepairs with Illumina, 100-500 basepairs with 454-sequencing). CARMA is a new software pipeline for the characterisation of species composition and the genetic potential of microbial samples using short, unassembled reads. RESULTS: In this paper, we introduce WebCARMA, a refined version of CARMA available as a web application for the taxonomic and functional classification of unassembled (ultra )short reads from metagenomic communities. In addition, we have analysed the applicability of ultra-short reads in metagenomics. CONCLUSIONS: We show that unassembled reads as short as 35 bp can be used for the taxonomic classification of a metagenome. The web application is freely available at http://webcarma.cebitec.uni-bielefeld.de. PMID- 20021648 TI - Comparing methods for identifying patients with heart failure using electronic data sources. AB - BACKGROUND: Accurately identifying heart failure (HF) patients from administrative claims data is useful for both research and quality of care efforts. Yet, there are few comparisons of the various claims data criteria (also known as claims signatures) for identifying HF patients. We compared various HF claim signatures to assess their relative accuracy. METHODS: In this retrospective study, we identified 4174 patients who received care from a large health system in southeast Michigan and who had >or=1 HF encounter between January 1, 2004 and December 31, 2005. Four hundred patients were chosen at random and a detailed chart review was performed to assess which met the Framingham HF criteria. The sample was divided into 300 subjects for derivation and 100 subjects for validation. Sensitivity, specificity,, and area under the curve (AUC) were determined for the various claim signatures. The criteria with the highest AUC were retested in the validation set. RESULTS: Of the 400 patients sampled, 65% met Framingham HF criteria, and 56% had at least one B-type Natriuretic Peptide (BNP) measurement. There was substantial variation between claims signatures in terms of sensitivity (range 15%-77%) and specificity (range 69%-100%). The best performing criteria in the derivation set was if patients met any one of the following: >or=2 HF encounters, any hospital discharge diagnosis of HF, or a BNP >or=200 pg/ml. These criteria showed a sensitivity of 76%, specificity of 75%, and AUC of 0.754 for meeting the Framingham HF criteria. This claims signature performed similarly in the validation set. CONCLUSION: Claim signatures for HF vary greatly in their relative sensitivity and specificity. These findings may facilitate efforts to identify HF patients for research and quality improvement efforts. PMID- 20021647 TI - Oral malignant melanomas and other head and neck neoplasms in Danish dogs--data from the Danish Veterinary Cancer Registry. AB - BACKGROUND: Head and neck cancers (HNC) are relatively common and often very serious diseases in both dogs and humans. Neoplasms originating in the head and neck region are a heterogeneous group. HNC often has an unfavourable prognosis and the proximity of the tissue structures renders extirpation of tumours with sufficient margins almost incompatible with preservation of functionality. In humans oral malignant melanoma (OMM) is extremely rare, but represents a particular challenge since it is highly aggressive as is the canine counterpart, which thus may be of interest as a spontaneous animal model. METHODS: Canine cases entered in the Danish Veterinary Cancer Registry (DVCR) from May 15th 2005 through February 29th 2008 were included in this study. Fisher's exact test was used to compare proportions of HNC in dogs and humans as well as proportions of surgically treated cases of OMM and squamous cell carcinomas (SCC). Also the proportions of benign and malignant neoplasms of different locations in dogs were compared using Fisher's exact test. RESULTS: A total of 1768 cases of neoplasias (679 malignant, 826 benign, 263 unknown) were submitted. Of all neoplasias HNC accounted for 7.2% (n = 128). Of these, 64 (50%) were malignant and 44 (34%) benign. The most common types of malignant neoplasia were SCC (18; 28% of malignant), OMM (13; 20% of malignant), soft tissue sarcoma (11; 17% of malignant) and adenocarcinoma (5; 11% of malignant). The most common types of benign neoplasms were adenoma (7; 16% of benign), polyps (6; 14% of benign) and fibroma (5; 11% of benign). CONCLUSIONS: In the current study, the proportion of neoplasia in the head and neck region in dogs in Denmark was similar to other canine studies and significantly more common than in humans with a large proportion of malignancies. Spontaneous HNC in dogs thus, may serve as a model for HNC in humans.Canine OMM is a spontaneous cancer in an outbred, immune competent large mammal population and could be a clinical model for OMM in humans. PMID- 20021649 TI - Factors associated with the desire for orthodontic treatment among Brazilian adolescents and their parents. AB - BACKGROUND: In the period of adolescence physical appearance takes on significant importance in the construction of personal identity, including one's relationship with one's own body. A variety of social, cultural, psychological and personal factors influences the self-perception of dental appearance and the decision to undergo orthodontic treatment. Adolescents who seek orthodontic treatment are concerned with improving their appearance and social acceptance. The aim of the present study was to determine factors associated to the desire for orthodontic treatment among Brazilian adolescents and their parents. METHODS: The sample consisted of 403 subjects aged 14 to 18 years, selected randomly from a population of 182,291 schoolchildren in the same age group. The outcome variable "desire for orthodontic treatment" was assessed through a questionnaire. Self perception of dental aesthetics was assessed using the Oral Aesthetic Subjective Impact Scale (OASIS) and the Dental Aesthetic Index (DAI) was used for clinical assessment. Statistical analysis involved the chi-square test as well as both simple and multiple logistic regression analyses. RESULTS: The majority (78%) of the Brazilian adolescents desired orthodontic treatment and 69% of the parents reported that their children were not in orthodontic treatment due to the high costs involved. There was significant association (p < or = 0.05) between the desire for orthodontic treatment and most types of malocclusion. However, there was no significant association between the desire for orthodontic treatment and the variables gender and age. CONCLUSIONS: The following were considered factors associated to the desire for treatment: upper anterior crowding > or = 2 mm and parents' perception of their child's need for treatment. PMID- 20021650 TI - Factors associated with tocolytic hospitalizations in Taiwan: evidence from a population-based and longitudinal study from 1997 to 2004. AB - BACKGROUND: The use of tocolytic hospitalization in antenatal care is controversial and worthy of more research. We investigated individual, institutional, and area factors that affect the use of tocolytic hospitalizations in Taiwan where fertility has rapidly declined. METHODS: Longitudinal data from the 1996 to 2004 National Health Insurance Research Database in Taiwan were used to identify tocolytic hospitalizations. The probit model was used to estimate factors associated with tocolytic hospitalizations. RESULTS: The decline in fertility was significantly associated with the probability of tocolytic hospitalizations. Several physician and institutional factors-including physician's age, hospital ownership, accreditation status, bed size, and teaching status-were also significantly correlated to the dependent variables. CONCLUSIONS: The provision of inpatient tocolysis is influenced not only by clinical considerations but also by physician, institutional, and area factors unrelated to clinical need. Fertility declines in Taiwan may have led obstetricians/gynecologists to provide more tocolysis to make up for their lost income. If the explanation is further validated, reimbursement policies may need to be reviewed to correct for overuse of inpatient tocolysis. The correlation could also be explained by the increasing use of artificial reproductive technologies and higher social value of newborns. In addition, the physician and institutional variations observed in the study indicate potential misuse of inpatient tocolysis that warrant further investigation. PMID- 20021651 TI - Teleconsultation service to improve healthcare in rural areas: acceptance, organizational impact and appropriateness. AB - BACKGROUND: Nowadays, new organisational strategies should be identified to improve primary care and its link with secondary care in terms of efficacy and timeliness of interventions thus preventing unnecessary hospital accesses and costs saving for the health system. The purpose of this study is to assess the effects of the use of teleconsultation by general practitioners in rural areas. METHODS: General practitioners were provided with a teleconsultation service from 2006 to 2008 to obtain a second opinion for cardiac, dermatological and diabetic problems. Access, acceptance, organisational impact, effectiveness and economics data were collected. Clinical and access data were systematically entered in a database while acceptance and organisational data were evaluated through ad hoc questionnaires. RESULTS: There were 957 teleconsultation contacts which resulted in access to health care services for 812 symptomatic patients living in 30 rural communities. Through the teleconsultation service, 48 general practitioners improved the appropriateness of primary care and the integration with secondary care. In fact, the level of concordance between intentions and consultations for cardiac problems was equal to 9%, in 86% of the cases the service entailed a saving of resources and in 5% of the cases, it improved the timeliness. 95% of the GPs considered the overall quality positively. For a future routine use of this service, trust in specialists, duration and workload of teleconsultations and reimbursement should be taken into account. CONCLUSIONS: Managerial and policy implications emerged mainly related to the support to GPs in the provision of high quality primary care and decision-making processes in promoting similar services. PMID- 20021652 TI - The expansion of amino-acid repeats is not associated to adaptive evolution in mammalian genes. AB - BACKGROUND: The expansion of amino acid repeats is determined by a high mutation rate and can be increased or limited by selection. It has been suggested that recent expansions could be associated with the potential of adaptation to new environments. In this work, we quantify the strength of this association, as well as the contribution of potential confounding factors. RESULTS: Mammalian positively selected genes have accumulated more recent amino acid repeats than other mammalian genes. However, we found little support for an accelerated evolutionary rate as the main driver for the expansion of amino acid repeats. The most significant predictors of amino acid repeats are gene function and GC content. There is no correlation with expression level. CONCLUSIONS: Our analyses show that amino acid repeat expansions are causally independent from protein adaptive evolution in mammalian genomes. Relaxed purifying selection or positive selection do not associate with more or more recent amino acid repeats. Their occurrence is slightly favoured by the sequence context but mainly determined by the molecular function of the gene. PMID- 20021653 TI - Discovering collectively informative descriptors from high-throughput experiments. AB - BACKGROUND: Improvements in high-throughput technology and its increasing use have led to the generation of many highly complex datasets that often address similar biological questions. Combining information from these studies can increase the reliability and generalizability of results and also yield new insights that guide future research. RESULTS: This paper describes a novel algorithm called BLANKET for symmetric analysis of two experiments that assess informativeness of descriptors. The experiments are required to be related only in that their descriptor sets intersect substantially and their definitions of case and control are consistent. From resulting lists of n descriptors ranked by informativeness, BLANKET determines shortlists of descriptors from each experiment, generally of different lengths p and q. For any pair of shortlists, four numbers are evident: the number of descriptors appearing in both shortlists, in exactly one shortlist, or in neither shortlist. From the associated contingency table, BLANKET computes Right Fisher Exact Test (RFET) values used as scores over a plane of possible pairs of shortlist lengths 12. BLANKET then chooses a pair or pairs with RFET score less than a threshold; the threshold depends upon n and shortlist length limits and represents a quality of intersection achieved by less than 5% of random lists. CONCLUSIONS: Researchers seek within a universe of descriptors some minimal subset that collectively and efficiently predicts experimental outcomes. Ideally, any smaller subset should be insufficient for reliable prediction and any larger subset should have little additional accuracy. As a method, BLANKET is easy to conceptualize and presents only moderate computational complexity. Many existing databases could be mined using BLANKET to suggest optimal sets of predictive descriptors. PMID- 20021654 TI - The INeS study: prevention of multiple pregnancies: a randomised controlled trial comparing IUI COH versus IVF e SET versus MNC IVF in couples with unexplained or mild male subfertility. AB - BACKGROUND: Multiple pregnancies are high risk pregnancies with higher chances of maternal and neonatal mortality and morbidity. In the past decades the number of multiple pregnancies has increased. This trend is partly due to the fact that women start family planning at an increased age, but also due to the increased use of ART.Couples with unexplained or mild male subfertility generally receive intrauterine insemination IUI with controlled hormonal stimulation (IUI COH). The cumulative pregnancy rate is 40%, with a 10% multiple pregnancy rate.This study aims to reveal whether alternative treatments such as IVF elective Single Embryo Transfer (IVF e SET) or Modified Natural Cycle IVF (MNC IVF) can reduce the number of multiple pregnancy rates, but uphold similar pregnancy rates as IUI COH in couples with mild male or unexplained subfertility. Secondly, the aim is to perform a cost effective analyses and assess treatment preference of these couples. METHODS/DESIGN: We plan a multicentre randomised controlled clinical trial in the Netherlands comparing six cycles of intra-uterine insemination with controlled ovarian hyperstimulation or six cycles of Modified Natural Cycle (MNC) IVF or three cycles with IVF-elective Single Embryo Transfer (eSET) plus cryo cycles within a time frame of 12 months.Couples with unexplained subfertility or mild male subfertility and a poor prognosis for treatment independent pregnancy will be included. Women with anovulatory cycles, severe endometriosis, double sided tubal pathology or serious endocrine illness will be excluded.Our primary outcome is the birth of a healthy singleton. Secondary outcomes are multiple pregnancy, treatment costs, and patient experiences in each treatment arm. The analysis will be performed according tot the intention to treat principle. We will test for non-inferiority of the three arms with respect to live birth. As we accept a 12.5% loss in pregnancy rate in one of the two IVF arms to prevent multiple pregnancies, we need 200 couples per arm (600 couples in total). DISCUSSION: Determining the safest and most cost-effective treatment will ensure optimal chances of pregnancy for subfertile couples with substantially diminished perinatal and maternal complications. Should patients find the most cost effective treatment acceptable or even preferable, this could imply the need for a world wide shift in the primary treatment. TRIAL REGISTRATION: Current Controlled Trials ISRCTN 52843371. PMID- 20021655 TI - RiskDiff: a web tool for the analysis of the difference due to risk and demographic factors for incidence or mortality data. AB - BACKGROUND: Analysing the observed differences for incidence or mortality of a particular disease between two different situations (such as time points, geographical areas, gender or other social characteristics) can be useful both for scientific or administrative purposes. From an epidemiological and public health point of view, it is of great interest to assess the effect of demographic factors in these observed differences in order to elucidate the effect of the risk of developing a disease or dying from it. The method proposed by Bashir and Esteve, which splits the observed variation into three components: risk, population structure and population size is a common choice at practice. RESULTS: A web-based application, called RiskDiff has been implemented (available at http://rht.iconcologia.net/riskdiff.htm), to perform this kind of statistical analyses, providing text and graphical summaries. Code from the implemented functions in R is also provided. An application to cancer mortality data from Catalonia is used for illustration. CONCLUSIONS: Combining epidemiological with demographical factors is crucial for analysing incidence or mortality from a disease, especially if the population pyramids show substantial differences. The tool implemented may serve to promote and divulgate the use of this method to give advice for epidemiologic interpretation and decision making in public health. PMID- 20021656 TI - Endangered edible orchids and vulnerable gatherers in the context of HIV/AIDS in the southern highlands of Tanzania. AB - BACKGROUND: Tanzania is a wild orchid biodiversity hotspot and has a high prevalence of HIV/AIDS. The wild orchids in the study are endemic and protected by the Convention on International Trade in Endangered Species. Every year, however, between 2.2 and 4.1 million orchid plants consumed in Zambia are estimated as originating from Tanzania. This research examines the differences between HIV/AIDS wild edible orchid gatherers and non-HIV/AIDS gatherers with regards to the frequency of gathering, salience in naming the various orchids, gathering knowledge acquisition and perceptions regarding the current state of abundance of the edible species. METHODS: Data was collected through interviews with 224 individuals in the Makete District of Tanzania close to the boarder of Zambia. Free-listings were conducted and Sutrup's Cultural Significance Index (CSI) constructed. The independent t-test was used to compare the differences in gathering frequencies between affected and non-affected gatherers. A multiple comparison of the 4 subgroups (affected adults and children, and non-affected adults and children) in gathering frequencies was done with a one way ANOVA test and its post hoc test. To examine the difference between affected and non affected gatherers difference in source of gathering knowledge, a chi square test was run. RESULTS: Forty two vernacular names of gathered orchid species were mentioned corresponding to 7 botanical species belongs to genera Disa, Satyrium, Habenaria, Eulophia and Roeperocharis. Ninety-seven percent of HIV/AIDS affected households state that orchid gathering is their primary economic activity compared to non-HIV/AIDS affected households at 9.7 percent. The HIV/AIDS affected gathered significantly more often than the non-affected. AIDS orphans, however, gathered most frequently. Gatherers perceive a decreasing trend of abundance of 6 of the 7 species. Gathering activities were mainly performed in age based peer groups. The results revealed a significant difference between affected and non-affected individuals in terms of their source of gathering knowledge. CONCLUSIONS: HIV/AIDS is related to increased reliance on the natural environment. This appears even more so for the most vulnerable, the AIDS orphaned children followed by HIV/AIDS widows. PMID- 20021657 TI - Population pharmacokinetics of artesunate and dihydroartemisinin following single and multiple-dosing of oral artesunate in healthy subjects. AB - BACKGROUND: The population pharmacokinetics of artesunate (AS) and its active metabolite dihydroartemisinin (DHA) were studied in healthy subjects receiving single- or multiple-dosing of AS orally either in combination with pyronaridine (PYR) or as a monotherapy with or without food. METHODS: Data from 118 concentration-time profiles arising from 91 healthy Korean subjects were pooled from four Phase I clinical studies. Subjects received 2-5 mg/kg of single- and multiple-dosing of oral AS either in combination with PYR or as a monotherapy with or without food. Plasma AS and DHA were measured simultaneously using a validated liquid chromatography- mass spectrometric method with a lower limit of quantification of 1 ng/mL for both AS and DHA. Nonlinear mixed-effect modelling was used to obtain the pharmacokinetic and variability (inter-individual and residual variability) parameter estimates. RESULTS: A novel parent-metabolite pharmacokinetic model consisting of a dosing compartment, a central compartment for AS, a central compartment and a peripheral compartment for DHA was developed. AS and DHA data were modelled simultaneously assuming stoichiometric conversion to DHA. AS was rapidly absorbed with a population estimate of absorption rate constant (Ka) of 3.85 h-1. The population estimates of apparent clearance (CL/F) and volume of distribution (V2/F) for AS were 1190 L/h with 36.2% inter individual variability (IIV) and 1210 L with 57.4% IIV, respectively. For DHA, the population estimates of apparent clearance (CLM/F) and central volume of distribution (V3/F) were 93.7 L/h with 28% IIV and 97.1 L with 30% IIV, respectively. The population estimates of apparent inter-compartmental clearance (Q/F) and peripheral volume of distribution (V4/F) for DHA were 5.74 L/h and 18.5 L, respectively. Intake of high-fat and high-caloric meal prior to the drug administration resulted in 84% reduction in Ka. Body weight impacted CLM/F, such that a unit change in weight resulted in 1.9-unit change in CLM/F in the same direction. CONCLUSIONS: A novel simultaneous parent-metabolite pharmacokinetic model with good predictive power was developed to study the population pharmacokinetics of AS and DHA in healthy subjects following single- and multiple dosing of AS with or without the presence of food. Food intake and weight were significant covariates for Ka and CLM/F, respectively. PMID- 20021658 TI - Human Immunodeficiency Virus and Human Papilloma Virus - why HPV-induced lesions do not spontaneously resolve and why therapeutic vaccination can be successful. AB - HIV and HPV can both cause chronic infections and are acquired during sexual contact. HIV infection results in a progressive loss of CD4+ T cells that is associated with an increased prevalence of HPV infections, type-specific persistence and an increase in HPV-associated malignancies. On the one hand this illustrates the important role of HPV-specific CD4+ helper T-cell immunity, on the other it shows the Achilles heel of the HPV-specific immune response. The use of highly active antiretroviral therapy (HAART) results in a rapid reduction of HIV and a reconstitution of systemic CD4+ T-cell levels. The use of HAART thus has the potential to raise immunity to HPV but to the surprise of many, the incidence of HPV-induced diseases has increased rather than declined since the introduction of HAART. Here, the knowledge on how HPV-induced diseases develop in the face of a non-compromised immune system will be used to explain why the effect of HAART on HPV-induced diseases is modest at best. Furthermore, exciting new data in the field of therapeutic vaccines against HPV will be discussed as this may form a more durable and clinically successful therapeutic approach for the treatment of HPV-induced high-grade lesions in HIV-positive subjects on HAART. PMID- 20021659 TI - Comparison of the receptor FGFRL1 from sea urchins and humans illustrates evolution of a zinc binding motif in the intracellular domain. AB - BACKGROUND: FGFRL1, the gene for the fifth member of the fibroblast growth factor receptor (FGFR) family, is found in all vertebrates from fish to man and in the cephalochordate amphioxus. Since it does not occur in more distantly related invertebrates such as insects and nematodes, we have speculated that FGFRL1 might have evolved just before branching of the vertebrate lineage from the other invertebrates (Beyeler and Trueb, 2006). RESULTS: We identified the gene for FGFRL1 also in the sea urchin Strongylocentrotus purpuratus and cloned its mRNA. The deduced amino acid sequence shares 62% sequence similarity with the human protein and shows conservation of all disulfides and N-linked carbohydrate attachment sites. Similar to the human protein, the S. purpuratus protein contains a histidine-rich motif at the C-terminus, but this motif is much shorter than the human counterpart. To analyze the function of the novel motif, recombinant fusion proteins were prepared in a bacterial expression system. The human fusion protein bound to nickel and zinc affinity columns, whereas the sea urchin protein barely interacted with such columns. Direct determination of metal ions by atomic absorption revealed 2.6 mole zinc/mole protein for human FGFRL1 and 1.7 mole zinc/mole protein for sea urchin FGFRL1. CONCLUSION: The FGFRL1 gene has evolved much earlier than previously assumed. A comparison of the intracellular domain between sea urchin and human FGFRL1 provides interesting insights into the shaping of a novel zinc binding domain. PMID- 20021661 TI - Autistic disorder associated with a paternally derived unbalanced translocation leading to duplication of chromosome 15pter-q13.2: a case report. AB - Autism spectrum disorders have been associated with maternally derived duplications that involve the imprinted region on the proximal long arm of chromosome 15. Here we describe a boy with a chromosome 15 duplication arising from a 3:1 segregation error of a paternally derived translocation between chromosome 15q13.2 and chromosome 9q34.12, which led to trisomy of chromosome 15pter-q13.2 and 9q34.12-qter. Using array comparative genome hybridization, we localized the breakpoints on both chromosomes and sequence homology suggests that the translocation arose from non-allelic homologous recombination involving the low copy repeats on chromosome 15. The child manifests many characteristics of the maternally-derived duplication chromosome 15 phenotype including developmental delays with cognitive impairment, autism, hypotonia and facial dysmorphisms with nominal overlap of the most general symptoms found in duplications of chromosome 9q34. This case suggests that biallelically expressed genes on proximal 15q contribute to the idic(15) autism phenotype. PMID- 20021662 TI - Developmental iodine deficiency resulting in hypothyroidism reduces hippocampal ERK1/2 and CREB in lactational and adolescent rats. AB - BACKGROUND: Developmental iodine deficiency (ID) leads to inadequate thyroid hormone that impairs learning and memory with an unclear mechanism. Here, we show that hippocampal extracellular signal-regulated kinase (ERK1/2) and cAMP response element-binding protein (CREB) are implicated in the impaired learning and memory in lactational and adolescent rat hippocampus following developmental ID and hypothyroidism. METHODS: Three developmental rat models were created by administrating dam rats with either iodine-deficient diet or propylthiouracil (PTU, 5 ppm or 15 ppm)-added drinking water from gestational day (GD) 6 till postnatal day (PN) 28. Then, the total and phorsporylated ERK1/2 and total and phorsporylated CREB in the hippocampus were detected with western blot on PN14, PN21, PN28 and PN42. RESULTS: The iodine-deficient and hypothyroid pups showed lower serum FT3 and FT4 levels, smaller body size, and delayed eyes opening. The mean number of surviving cells in the hippocampus of the iodine-deficient and 15 ppm PTU-treated rats was significantly reduced compared to controls (P < 0.05). Iodine-deficient and 15 ppm PTU-treatment groups demonstrated significantly lower level of total and phosphorylated ERK1/2 and CREB than the controls on PN14, PN21 and PN28 (P < 0.05, respectively). The reduction of ERK1/2 and CREB was not reversible with the restoration of serum thyroid hormone concentrations on PN42. CONCLUSIONS: Developmental ID and hypothyroidism down-regulate hippocampal ERK1/2 and CREB in lactational and adolescent rats. PMID- 20021660 TI - Multilocus sequence typing supports the hypothesis that Ochrobactrum anthropi displays a human-associated subpopulation. AB - BACKGROUND: Ochrobactrum anthropi is a versatile bacterial species with strains living in very diverse habitats. It is increasingly recognized as opportunistic pathogen in hospitalized patients. The population biology of the species particularly with regard to the characteristics of the human isolates is being investigated. To address this issue, we proposed a polyphasic approach consisting in Multi-Locus Sequence Typing (MLST), multi-locus phylogeny, genomic-based fingerprinting by pulsed-field gel electrophoresis (PFGE) and antibiotyping. RESULTS: We tested a population of 70 O. anthropi clinical (n = 43) and environmental (n = 24) isolates as well as the type strain O. anthropi ATCC49188T and 2 strains of Ochrobactrum lupini and Ochrobactrum cytisi isolated from plant nodules. A Multi-Locus Sequence Typing (MLST) scheme for O. anthropi is proposed here for the first time. It was based on 7 genes (3490 nucleotides) evolving mostly by neutral mutations. The MLST approach suggested an epidemic population structure. A major clonal complex corresponded to a human-associated lineage since it exclusively contained clinical isolates. Genomic fingerprinting separated isolates displaying the same sequence type but it did not detect a population structure that could be related to the origin of the strains. None of the molecular method allowed the definition of particular lineages associated to the host-bacteria relationship (carriage, colonisation or infection). Antibiotyping was the least discriminative method. CONCLUSION: The results reveal a human-associated subpopulation in our collection of strains. The emergence of this clonal complex was probably not driven by the antibiotic selective pressure. Therefore, we hypothesise that the versatile species O. anthropi could be considered as a human-specialized opportunistic pathogen. PMID- 20021663 TI - Reduced expression of cenp-e in human hepatocellular carcinoma. AB - BACKGROUND: CENP-E, one of spindle checkpoint proteins, plays a crucial role in the function of spindle checkpoint. Once CENP-E expression was interrupted, the chromosomes can not separate procedurally, and may result in aneuploidy which is a hallmark of most solid cancers, such as hepatocellular carcinoma (HCC). We investigate the expression of CENP-E in human hepatocellular carcinoma,. and analyze the effect of low CENP-E expression on chromosome separation in normal liver cell line (LO2). METHODS: We determined its levels in HCC and para cancerous tissues, human hepatocellular carcinoma-derived cell line (HepG2) and LO2 cell line using real time quantitative PCR (QPCR) and Western blot. Further to know whether reduction in CENP-E expression impairs chromosomes separation in LO2 cells. we knocked down CENP-E using shRNA expressing vector and then count the aneuploid in LO2 cells using chromosomal counts assay. RESULTS: We found that both CENP-E mRNA and protein levels were significantly reduced in HCC tissues and HepG2 cells compared with para-cancerous tissues and LO2 cells, respectively. A significantly-increased proportion of aneuploid in these down-knocked LO2 cells compared with those treated with control shRNA vector. CONCLUSIONS: Together with other results, these results reveal that CENP-E expression was reduced in human HCC tissue, and low CENP-E expression result in aneuploidy in LO2 cells. PMID- 20021665 TI - Algorithms for locating extremely conserved elements in multiple sequence alignments. AB - BACKGROUND: In 2004, Bejerano et al. announced the startling discovery of hundreds of "ultraconserved elements", long genomic sequences perfectly conserved across human, mouse, and rat. Their announcement stimulated a flurry of subsequent research. RESULTS: We generalize the notion of ultraconserved element in a natural way from extraordinary human-rodent conservation to extraordinary conservation over an arbitrary set of species. We call these "Extremely Conserved Elements". There is a linear time algorithm to find all such Extremely Conserved Elements in any multiple sequence alignment, provided that the conservation is required to be across all the aligned species. For the general case of conservation across an arbitrary subset of the aligned species, we show that the question of whether there exists an Extremely Conserved Element is NP-complete. We illustrate the linear time algorithm by cataloguing all 177 Extremely Conserved Elements in the currently available 44-vertebrate whole-genome alignment, and point out some of the characteristics of these elements. CONCLUSIONS: The NP-completeness in the case of conservation across an arbitrary subset of the aligned species implies that it is unlikely an efficient algorithm exists for this general case. Despite this fact, for the interesting case of conservation across all or most of the aligned species, our algorithm is efficient enough to be practical. The 177 Extremely Conserved Elements that we catalog demonstrate many of the characteristics of the original ultraconserved elements of Bejerano et al. PMID- 20021664 TI - Predictors of metabolic monitoring among schizophrenia patients with a new episode of second-generation antipsychotic use in the Veterans Health Administration. AB - BACKGROUND: To examine the baseline metabolic monitoring (MetMon) for second generation antipsychotics (SGA) among patients with schizophrenia in the Veterans Integrated Service Network (VISN) 16 of the Veterans Health Administration (VHA). METHODS: VISN16 electronic medical records for 10/2002-08/2005 were used to identify patients with schizophrenia who received a new episode of SGA treatment after 10/2003, in which the VISN 16 baseline MetMon program was implemented. Patients who underwent MetMon (MetMon+: either blood glucose or lipid testing records) were compared with patients who did not (MetMon-), on patient characteristics and resource utilization in the year prior to index treatment episode. A parsimonious logistic regression was used to identify predictors for MetMon+ with adjusted odds ratios (OR) and 95% confidence intervals (CI). RESULTS: Out of 4,709 patients, 3,568 (75.8%) underwent the baseline MetMon. Compared with the MetMon- group, the MetMon+ patients were found more likely to have baseline diagnoses or mediations for diabetes (OR [CI]: 2.336 [1.846 2.955]), dyslipidemia (2.439 [2.029-2.932]), and hypertension (1.497 [1.287 1.743]), substance use disorders (1.460 [1.257-1.696]), or to be recorded as obesity (2.052 [1.724-2.443]). Increased likelihood for monitoring were positively associated with number of antipsychotics during the previous year (FGA: 1.434 [1.129-1.821]; SGA: 1.503 [1.290-1.751]). Other significant predictors for monitoring were more augmentation episodes (1.580 [1.145-2.179]), more outpatient visits (1.007 [1.002-1.013])), hospitalization days (1.011 [1.007 1.015]), and longer duration of antipsychotic use (1.001 [1.001-1.001]). Among the MetMon+ group, approximately 38.9% patient had metabolic syndrome. DISCUSSION: This wide time window of 180 days, although congruent with the VHA guidelines for the baseline MetMon process, needs to be re-evaluated and narrowed down, so that optimally the monitoring event occurs at the time of receiving a new episode of SGA treatment. Future research will examine whether or not patients prescribed an SGA are assessed for metabolic syndrome following the index episode of antipsychotic therapy, and whether or not such baseline and follow-up monitoring programs in routine care are cost-effective. CONCLUSION: The baseline MetMon has been performed for a majority of the VISN 16 patients with schizophrenia prior to index SGA over the study period. Compared with MetMon- group, MetMon+ patients were more likely to be obese and manifest a more severe illness profile. PMID- 20021666 TI - Tetrabenazine as anti-chorea therapy in Huntington disease: an open-label continuation study. Huntington Study Group/TETRA-HD Investigators. AB - BACKGROUND: Tetrabenazine (TBZ) selectively depletes central monoamines by reversibly binding to the type-2 vesicular monoamine transporter. A previous double blind study in Huntington disease (HD) demonstrated that TBZ effectively suppressed chorea, with a favorable short-term safety profile (Neurology 2006;66:366-372). The objective of this study was to assess the long-term safety and effectiveness of TBZ for chorea in HD. METHODS: Subjects who completed the 13 week, double blind protocol were invited to participate in this open label extension study for up to 80 weeks. Subjects were titrated to the best individual dose or a maximum of 200 mg/day. Chorea was assessed using the Total Maximal Chorea (TMC) score from the Unified Huntington Disease Rating Scale. RESULTS: Of the 75 participants, 45 subjects completed 80 weeks. Three participants terminated due to adverse events (AEs) including depression, delusions with associated previous suicidal behavior, and vocal tics. One subject died due to breast cancer. The other 26 subjects chose not to continue on with each ensuing extension for various reasons. When mild and unrelated AEs were excluded, the most commonly reported AEs (number of subjects) were sedation/somnolence (18), depressed mood (17), anxiety (13), insomnia (10), and akathisia (9). Parkinsonism and dysarthria [corrected] scores were significantly increased at week 80 compared to baseline. At week 80, chorea had significantly improved from baseline with a mean reduction in the TMC score of 4.6 (SD 5.5) units. The mean dosage at week 80 was 63.4 mg (range 12.5-175 mg). CONCLUSIONS: TBZ effectively suppresses HD-related chorea for up to 80 weeks. Patients treated chronically with TBZ should be monitored for parkinsonism, dysphagia and other side effects including sleep disturbance, depression, anxiety, and akathisia. TRIAL REGISTRATION: Clinicaltrials.gov registration number (initial study): NCT00219804. PMID- 20021667 TI - Short-term cultured, interleukin-15 differentiated dendritic cells have potent immunostimulatory properties. AB - BACKGROUND: Optimization of the current dendritic cell (DC) culture protocol in order to promote the therapeutic efficacy of DC-based immunotherapy is warranted. Alternative differentiation of monocyte-derived DCs using granulocyte macrophage colony-stimulating factor (GM-CSF) and interleukin (IL)-15 has been propagated as an attractive strategy in that regard. The applicability of these so-called IL-15 DCs has not yet been firmly established. We therefore developed a novel pre clinical approach for the generation of IL-15 DCs with potent immunostimulatory properties. METHODS: Human CD14+ monocytes were differentiated with GM-CSF and IL 15 into immature DCs. Monocyte-derived DCs, conventionally differentiated in the presence of GM-CSF and IL-4, served as control. Subsequent maturation of IL-15 DCs was induced using two clinical grade maturation protocols: (i) a classic combination of pro-inflammatory cytokines (tumor necrosis factor-alpha, IL-1beta, IL-6, prostaglandin E2) and (ii) a Toll-like receptor (TLR)7/8 agonist-based cocktail (R-848, interferon-gamma, TNF-alpha and prostaglandin E2). In addition, both short-term (2-3 days) and long-term (6-7 days) DC culture protocols were compared. The different DC populations were characterized with respect to their phenotypic profile, migratory properties, cytokine production and T cell stimulation capacity. RESULTS: The use of a TLR7/8 agonist-based cocktail resulted in a more optimal maturation of IL-15 DCs, as reflected by the higher phenotypic expression of CD83 and costimulatory molecules (CD70, CD80, CD86). The functional superiority of TLR7/8-activated IL-15 DCs over conventionally matured IL-15 DCs was evidenced by their (i) higher migratory potential, (ii) advantageous cytokine secretion profile (interferon-gamma, IL-12p70) and (iii) superior capacity to stimulate autologous, antigen-specific T cell responses after passive peptide pulsing. Aside from a less pronounced production of bioactive IL-12p70, short-term versus long-term culture of TLR7/8-activated IL-15 DCs resulted in a migratory profile and T cell stimulation capacity that was in favour of short-term DC culture. In addition, we demonstrate that mRNA electroporation serves as an efficient antigen loading strategy of IL-15 DCs. CONCLUSIONS: Here we show that short-term cultured and TLR7/8-activated IL-15 DCs fulfill all pre-clinical prerequisites of immunostimulatory DCs. The results of the present study might pave the way for the implementation of IL-15 DCs in immunotherapy protocols. PMID- 20021668 TI - Evolution of DNA ligases of nucleo-cytoplasmic large DNA viruses of eukaryotes: a case of hidden complexity. AB - BACKGROUND: Eukaryotic Nucleo-Cytoplasmic Large DNA Viruses (NCLDV) encode most if not all of the enzymes involved in their DNA replication. It has been inferred that genes for these enzymes were already present in the last common ancestor of the NCLDV. However, the details of the evolution of these genes that bear on the complexity of the putative ancestral NCLDV and on the evolutionary relationships between viruses and their hosts are not well understood. RESULTS: Phylogenetic analysis of the ATP-dependent and NAD-dependent DNA ligases encoded by the NCLDV reveals an unexpectedly complex evolutionary history. The NAD-dependent ligases are encoded only by a minority of NCLDV (including mimiviruses, some iridoviruses and entomopoxviruses) but phylogenetic analysis clearly indicated that all viral NAD-dependent ligases are monophyletic. Combined with the topology of the NCLDV tree derived by consensus of trees for universally conserved genes suggests that this enzyme was represented in the ancestral NCLDV. Phylogenetic analysis of ATP dependent ligases that are encoded by chordopoxviruses, most of the phycodnaviruses and Marseillevirus failed to demonstrate monophyly and instead revealed an unexpectedly complex evolutionary trajectory. The ligases of the majority of phycodnaviruses and Marseillevirus seem to have evolved from bacteriophage or bacterial homologs; the ligase of one phycodnavirus, Emiliana huxlei virus, belongs to the eukaryotic DNA ligase I branch; and ligases of chordopoxviruses unequivocally cluster with eukaryotic DNA ligase III. CONCLUSIONS: Examination of phyletic patterns and phylogenetic analysis of DNA ligases of the NCLDV suggest that the common ancestor of the extant NCLDV encoded an NAD-dependent ligase that most likely was acquired from a bacteriophage at the early stages of evolution of eukaryotes. By contrast, ATP-dependent ligases from different prokaryotic and eukaryotic sources displaced the ancestral NAD dependent ligase at different stages of subsequent evolution. These findings emphasize complex routes of viral evolution that become apparent through detailed phylogenomic analysis but not necessarily in reconstructions based on phyletic patterns of genes. REVIEWERS: This article was reviewed by: Patrick Forterre, George V. Shpakovski, and Igor B. Zhulin. PMID- 20021669 TI - Assessing the utility of whole-genome amplified serum DNA for array-based high throughput genotyping. AB - BACKGROUND: Whole genome amplification (WGA) offers new possibilities for genome wide association studies where limited DNA samples have been collected. This study provides a realistic and high-precision assessment of WGA DNA genotyping performance from 20-year old archived serum samples using the Affymetrix Genome Wide Human SNP Array 6.0 (SNP6.0) platform. RESULTS: Whole-genome amplified (WGA) DNA samples from 45 archived serum replicates and 5 fresh sera paired with non amplified genomic DNA were genotyped in duplicate. All genotyped samples passed the imposed QC thresholds for quantity and quality. In general, WGA serum DNA samples produced low call rates (45.00 +/- 2.69%), although reproducibility for successfully called markers was favorable (concordance = 95.61 +/- 4.39%). Heterozygote dropouts explained the majority (>85% in technical replicates, 50% in paired genomic/serum samples) of discordant results. Genotyping performance on WGA serum DNA samples was improved by implementation of Corrected Robust Linear Model with Maximum Likelihood Classification (CRLMM) algorithm but at the loss of many samples which failed to pass its quality threshold. Poor genotype clustering was evident in the samples that failed the CRLMM confidence threshold. CONCLUSIONS: We conclude that while it is possible to extract genomic DNA and subsequently perform whole-genome amplification from archived serum samples, WGA serum DNA did not perform well and appeared unsuitable for high-resolution genotyping on these arrays. PMID- 20021670 TI - Using mechanistic Bayesian networks to identify downstream targets of the sonic hedgehog pathway. AB - BACKGROUND: The topology of a biological pathway provides clues as to how a pathway operates, but rationally using this topology information with observed gene expression data remains a challenge. RESULTS: We introduce a new general purpose analytic method called Mechanistic Bayesian Networks (MBNs) that allows for the integration of gene expression data and known constraints within a signal or regulatory pathway to predict new downstream pathway targets. The MBN framework is implemented in an open-source Bayesian network learning package, the Python Environment for Bayesian Learning (PEBL). We demonstrate how MBNs can be used by modeling the early steps of the sonic hedgehog pathway using gene expression data from different developmental stages and genetic backgrounds in mouse. Using the MBN approach we are able to automatically identify many of the known downstream targets of the hedgehog pathway such as Gas1 and Gli1, along with a short list of likely targets such as Mig12. CONCLUSIONS: The MBN approach shown here can easily be extended to other pathways and data types to yield a more mechanistic framework for learning genetic regulatory models. PMID- 20021671 TI - Gene expression relationship between prostate cancer cells of Gleason 3, 4 and normal epithelial cells as revealed by cell type-specific transcriptomes. AB - BACKGROUND: Prostate cancer cells in primary tumors have been typed CD10-/CD13 /CD24hi/CD26+/CD38lo/CD44-/CD104-. This CD phenotype suggests a lineage relationship between cancer cells and luminal cells. The Gleason grade of tumors is a descriptive of tumor glandular differentiation. Higher Gleason scores are associated with treatment failure. METHODS: CD26+ cancer cells were isolated from Gleason 3+3 (G3) and Gleason 4+4 (G4) tumors by cell sorting, and their gene expression or transcriptome was determined by Affymetrix DNA array analysis. Dataset analysis was used to determine gene expression similarities and differences between G3 and G4 as well as to prostate cancer cell lines and histologically normal prostate luminal cells. RESULTS: The G3 and G4 transcriptomes were compared to those of prostatic cell types of non-cancer, which included luminal, basal, stromal fibromuscular, and endothelial. A principal components analysis of the various transcriptome datasets indicated a closer relationship between luminal and G3 than luminal and G4. Dataset comparison also showed that the cancer transcriptomes differed substantially from those of prostate cancer cell lines. CONCLUSIONS: Genes differentially expressed in cancer are potential biomarkers for cancer detection, and those differentially expressed between G3 and G4 are potential biomarkers for disease stratification given that G4 cancer is associated with poor outcomes. Differentially expressed genes likely contribute to the prostate cancer phenotype and constitute the signatures of these particular cancer cell types. PMID- 20021672 TI - Perceptions of hookah smoking harmfulness: predictors and characteristics among current hookah users. AB - INTRODUCTION: Tobacco cigarette smoking a well-known cause of cancer and other diseases. Hookah smoking is another form of tobacco use that has rapidly spread in the United State and Europe. This study assessed beliefs about the harmfulness of smoking hookah. METHODS: We surveyed hookah users in all cafes that provided hookah to its customers in downtown San Diego, California and nearby areas. A total of 235 hookah users participated in this study. RESULTS: Average age of study participants was 22 years, 57% were males, and 72% were not cigarette smokers. Whites were more likely to use hookah than the other ethnic groups (33%), older hookah users (26-35 years) were mostly males, and mint flavor of hookah tobacco was the most popular among a wide variety of flavors (23%). There was no significant difference in gender in relation to the wrong perception that hookah is less harmful than cigarettes, but those of Asian ethnicity were much less likely than other ethnic groups to believe that hookah is less harmful than cigarettes. More frequent users of hookah were more likely to believe that hookah is less harmful than cigarettes. The majority of hookah users (58.3%) believe hookah is less harmful than cigarette smoking. DISCUSSION: Compared to cigarettes, there appears to be a lack of knowledge about the harmfulness of smoking hookah among users regardless of their demographic background. Education about the harmfulness of smoking hookah and policies to limit its use should be implemented to prevent the spread of this new form of tobacco use. PMID- 20021673 TI - Pharmacology as a foreign language: a preliminary evaluation of podcasting as a supplementary learning tool for non-medical prescribing students. AB - BACKGROUND: Nurses and other health professionals in the U.K. can gain similar prescribing rights to doctors by undertaking a non-medical prescribing course. Non-medical prescribing students must have a thorough understanding of the pharmacology of prescribing to ensure safe practice. Pharmacology education at this level is complicated by the variation in students' prior subject knowledge of, and anxiety about, the subject. The recent advances in technology, particularly the potential for mobile learning, provide increased opportunities for students to familiarise themselves with lecture materials and hence promote understanding. The objective of this study was therefore to evaluate both the subjective (student perception) and objective (student use and exam results) usefulness of podcasts of pharmacology lectures which were provided as an extra learning tool to two cohorts (n = 69) of non-medical prescribing students. METHODS: The podcasts were made available to students through the virtual learning environment WebCT. Use of podcasts by two successive cohorts of nurse prescribing students (n = 69) was tracked through WebCT. Survey data, which was collected from 44 of these students, investigated patterns of/reasons for podcast use and perceived usefulness of podcasts as a learning tool. Of these 69 students, 64 completed the pharmacology exam. In order to examine any impact of podcasts on student knowledge, their exam results were compared with those of two historical cohorts who did not have access to podcasts (n = 70). RESULTS: WebCT tracking showed that 91% of students accessed at least one podcast. 93% of students used the podcasts to revisit a lecture, 85% used podcasts for revision, and 61% used the podcasts when they had a specific question. Only 22% used the podcasts because they had missed a pharmacology session. Most students (81%) generally listened to the entire podcast rather than specific sections and most (73%) used them while referring to their lecture handouts. The majority of students found the podcasts helpful as a learning tool, as a revision aid and in promoting their understanding of the subject. Evaluation of the range of marks obtained, mode mark and mean mark suggested improved knowledge in students with access to podcasts compared to historical cohorts of students who did not have access to pharmacology podcasts. CONCLUSIONS: The results of this study suggest that non-medical prescribing students utilised podcasts of pharmacology lectures, and have found the availability of these podcasts helpful for their learning. Exam results indicate that the availability of podcasts was also associated with improved exam performance. PMID- 20021674 TI - Atypical presentation of angiosarcoma of the scalp in the setting of human immunodeficiency virus (HIV). AB - BACKGROUND: Angiosarcoma of the head and neck is an uncommon, aggressive malignant entity most commonly found in elderly Caucasian males. We present a case in a young black female with co-existing HIV. The atypical gender, age and race of the patient reflect the unusual clinical presentation of this case of angiosarcoma, attributable to the patient's HIV status. CASE PRESENTATION: A 22 year old patient presented with a large unresectable lesion over the occiput with surrounding ulceration, satellite lesions and associated lymphadenopathy. She is HIV-infected with a CD4 count of 360 cells/microl. She was not on antiretroviral treatment based on South African treatment guidelines advocating antiretroviral treatment when the CD4 count is below 200 cells/microl, in the absence of other AIDS-defining illnesses.The patient was treated with a course of ifosfamide and anthracyline based chemotherapy. Disease progression was noted on chemotherapy and she was subsequently palliated with a course of radiotherapy. She had a satisfactory response with an improvement in local symptoms. She is currently receiving symptomatic care. CONCLUSIONS: South Africa is at the epicenter of the HIV epidemic. Consequently, the management of patients in the field of oncology in our clinical practice is often burdened with malignancies manifesting with an atypical disease presentation and clinical course. PMID- 20021675 TI - Dry needling and exercise for chronic whiplash - a randomised controlled trial. AB - BACKGROUND: Chronic whiplash is a common and costly problem. Sensory hypersensitivity is a feature of chronic whiplash that is associated with poor responsiveness to physical treatments such as exercise. Modalities such as dry needling have shown some capacity to modulate sensory hypersensitivity, suggesting that when combined with advice and exercise, such an approach may be more effective in the management of chronic whiplash. The primary aim of this project is to investigate the effectiveness of dry-needling, advice and exercise for chronic whiplash. METHOD/DESIGN: A double-blind randomised controlled trial will be conducted. 120 participants with chronic whiplash, grade II will be randomised to receive either 1) dry-needling, advice and exercise or 2) sham dry needling, advice and exercise. All participants will receive an educational booklet on whiplash. Participants who are randomised to Group 1 will receive 6 treatments of combined dry-needling and exercise delivered in the first 3 weeks of the 6 week program, and 4 treatments of exercise only in the last 3 weeks of the program. Participants randomised to Group 2 will receive an identical protocol, except that a sham dry-needling technique will be used instead of dry needling. The primary outcome measures are the Neck Disability Index (NDI) and participants' perceived recovery. Outcomes will be measured at 6, 12, 24 and 52 weeks after randomization by an assessor who is blind to the group allocation of the participants. In parallel, an economic analysis will be conducted. DISCUSSION: This trial will utilise high quality trial methodologies in accordance with CONSORT guidelines. The successful completion of this trial will provide evidence of the effectiveness and cost-effectiveness of a combined treatment approach for the management of chronic whiplash. TRIAL REGISTRATION: ACTRN12609000470291. PMID- 20021676 TI - Gamma Knife radiosurgery for vestibular schwannoma: case report and review of the literature. AB - Vestibular schwannomas, also called acoustic neuromas, are benign tumors of the vestibulocochlear nerve. Patients with these tumours almost always present with signs of hearing loss, and many also experience tinnitus, vertigo, and equilibrium problems. Following diagnosis with contrast enhanced MRI, patients may choose to observe the tumour with subsequent scans or seek active treatment in the form of microsurgery, radiosurgery, or radiotherapy. Unfortunately, definitive guidelines for treating vestibular schwannomas are lacking, because of insufficient evidence comparing the outcomes of therapeutic modalities.We present a contemporary case report, describing the finding of a vestibular schwannoma in a patient who presented with dizziness and a "clicking" sensation in the ear, but no hearing deficit. Audible clicking is a symptom that, to our knowledge, has not been associated with vestibular schwannoma in the literature. We discuss the diagnosis and patient's decision-making process, which led to treatment with Gamma Knife radiosurgery. Treatment resulted in an excellent radiographic response and complete hearing preservation. This case highlights an atypical presentation of vestibular schwannoma, associated with audible "clicks" and normal hearing. We also provide a concise review of the available literature on modern vestibular schwannoma treatment, which may be useful in guiding treatment decisions. PMID- 20021677 TI - A modified QuickDASH-9 provides a valid outcome instrument for upper limb function. AB - BACKGROUND: The 30-item Disabilities Arm Shoulder and Hand (DASH) questionnaire was introduced to facilitate assessment of upper limb functional limitations. To improve practicality and eliminate item redundancy a modified instrument was needed. The 11-item QuickDASH was developed to fulfil these requirements and translated into several languages. However, prospective investigations of psychometric and practical characteristics are limited. No published study investigated readability or used concurrent validation with a standardized upper limb criterion measure. The validity of the QuickDASH has been questioned as the results for factor structure are conflicting, and the English-language version has not yet had factor structure reported. A shortened 9-item version, the QuickDASH-9, that addresses these issues is proposed. METHODS: This two-stage observational study assessed the psychometric and practical characteristics of the QuickDASH and the extracted QuickDASH-9. The Upper Limb Functional Index (ULFI) was the criterion standard in both stages. Stage 1, calibration, reanalyzed extracted QuickDASH and QuickDASH-9 responses from a previous prospective study, by the authors, of the 30-item DASH (n = 137). Stage 2, prospective validation, investigated the QuickDASH through repeated measures in consecutive upper limb musculoskeletal participants' consulting for physical therapy in Australia (n = 67). The QuickDASH and extracted QuickDASH-9 data from both stages was analyzed and compared for psychometric properties, practical characteristics and factor structure. RESULTS: The proposed QuickDASH-9 had a unidimensional structure, high reliability (ICC 2:1, r = 0.92), internal consistency (alpha = 0.93) and responsiveness (ES = 1.05). It correlated highly with both the DASH (r = 0.97), QuickDASH (r = 0.99) and ULFI criterion (r = 0.85). QuickDASH-9 missing responses reduced to 3.5% from 26% in the QuickDASH. Completion and scoring time was 134 +/- 56 seconds and required a computational aid. The QuickDASH demonstrated a bidimensional structure making it invalid. The QuickDASH-9 summary performance was measured on the 'Measurement of Outcome Measures' at 88% and on the 'Bot' clinimetric scale at 75%. CONCLUSIONS: The proposed QuickDASH-9 had a unidimensional structure and similar psychometric precision to the full-length DASH with improved practicality and completion time. The QuickDASH was invalid as its bidimensional structure made a single summated score inappropriate. The QuickDASH-9 offers a future direction for ongoing use of the QuickDASH concept. PMID- 20021678 TI - Evaluation of self-reported ethnicity in a case-control population: the stroke prevention in young women study. AB - BACKGROUND: Population-based association studies are used to identify common susceptibility variants for complex genetic traits. These studies are susceptible to confounding from unknown population substructure. Here we apply a model-based clustering approach to our case-control study of stroke among young women to examine if self-reported ethnicity can serve as a proxy for genetic ancestry. FINDINGS: A population-based case-control study of stroke among women aged 15-49 identified 361 cases of first ischemic stroke and 401 age-comparable control subjects. Thirty single nucleotide polymorphisms (SNPs) throughout the genome unrelated to stroke risk and with established ancestry-based allele frequency differences were genotyped in all participants. The Structure program was used to iteratively evaluate for K = 1 to 5 potential genetic-based subpopulations. Evaluating the population as a whole, the Structure output plateaued at K = 2 clusters. 98% of self-reported Caucasians had an estimated probability >/=50% of belonging to Cluster 1, while 94% of self-reported African-Americans had an estimated probability >/=50% of belonging to Cluster 2. Stratifying the participants by self-reported ethnicity and repeating the analyses revealed the presence of two clusters among Caucasians, suggesting that potential substructure may exist. CONCLUSIONS: Among our combined sample of African-American and Caucasian participants there is no large unknown subpopulation and self-reported ethnicity can serve as a proxy for genetic ancestry. Ethnicity-specific analyses indicate that population substructure may exist among the Caucasian participants indicating that further studies are warranted. PMID- 20021679 TI - Cellular injury and neuroinflammation in children with chronic intractable epilepsy. AB - OBJECTIVE: To elucidate the presence and potential involvement of brain inflammation and cell death in neurological morbidity and intractable seizures in childhood epilepsy, we quantified cell death, astrocyte proliferation, microglial activation and cytokine release in brain tissue from patients who underwent epilepsy surgery. METHODS: Cortical tissue was collected from thirteen patients with intractable epilepsy due to focal cortical dysplasia (6), encephalomalacia (5), Rasmussen's encephalitis (1) or mesial temporal lobe epilepsy (1). Sections were processed for immunohistochemistry using markers for neuron, astrocyte, microglia or cellular injury. Cytokine assay was performed on frozen cortices. Controls were autopsy brains from eight patients without history of neurological diseases. RESULTS: Marked activation of microglia and astrocytes and diffuse cell death were observed in epileptogenic tissue. Numerous fibrillary astrocytes and their processes covered the entire cortex and converged on to blood vessels, neurons and microglia. An overwhelming number of neurons and astrocytes showed DNA fragmentation and its magnitude significantly correlated with seizure frequency. Majority of our patients with abundant cell death in the cortex have mental retardation. IL-1beta, IL-8, IL-12p70 and MIP-1beta were significantly increased in the epileptogenic cortex; IL-6 and MCP-1 were significantly higher in patients with family history of epilepsy. CONCLUSIONS: Our results suggest that active neuroinflammation and marked cellular injury occur in pediatric epilepsy and may play a common pathogenic role or consequences in childhood epilepsy of diverse etiologies. Our findings support the concept that immunomodulation targeting activated microglia and astrocytes may be a novel therapeutic strategy to reduce neurological morbidity and prevent intractable epilepsy. PMID- 20021680 TI - Flux-sum analysis: a metabolite-centric approach for understanding the metabolic network. AB - BACKGROUND: Constraint-based flux analysis of metabolic network model quantifies the reaction flux distribution to characterize the state of cellular metabolism. However, metabolites are key players in the metabolic network and the current reaction-centric approach may not account for the effect of metabolite perturbation on the cellular physiology due to the inherent limitation in model formulation. Thus, it would be practical to incorporate the metabolite states into the model for the analysis of the network. RESULTS: Presented herein is a metabolite-centric approach of analyzing the metabolic network by including the turnover rate of metabolite, known as flux-sum, as key descriptive variable within the model formulation. By doing so, the effect of varying metabolite flux sum on physiological change can be simulated by resorting to mixed integer linear programming. From the results, we could classify various metabolite types based on the flux-sum profile. Using the iAF1260 in silico metabolic model of Escherichia coli, we demonstrated that this novel concept complements the conventional reaction-centric analysis. CONCLUSIONS: Metabolite flux-sum analysis elucidates the roles of metabolites in the network. In addition, this metabolite perturbation analysis identifies the key metabolites, implicating practical application which is achievable through metabolite flux-sum manipulation in the areas of biotechnology and biomedical research. PMID- 20021681 TI - Supracricoid partial laryngectomy with cricohyoidoepiglottopexy in patients with radiation therapy failure. AB - BACKGROUND: To assess functional results, complications, and success of larynx preservation in patients with recurrent squamous cell carcinoma after radiotherapy. METHODS: From a database of 40 patients who underwent supracricoid partial laryngectomy (SCPL) with cricohyoidoepiglottopexy (CHEP) from June 2001 to April 2006, eight patients were treated previously with radiotherapy due to squamous cell carcinoma of the glottic region and were treated for recurrence at the site of the primary cancer. RESULTS: SCPL with CHEP was performed in six men and two women with a mean age of 67 years due to recurrence and/or persistence at a mean time of 30 months postradiotherapy (in case #8 after concomitant chemoradiotherapy). Bilateral neck dissection at levels II-V was performed in six patients. Only case #8 presented metastasis in one node. In case #5, Delphian node was positive. It was possible to preserve both arytenoids in five cases. Definitive surgical margins were negative. Complications were encountered in seven patients. Follow-up was on average 44 months (range: 20-67 months). Organ preservation in this series was 75%, and local control was 87%. Overall 5-year survival was 50%. CONCLUSIONS: In selected patient with persistence and/or recurrence after radiotherapy due to cancer of the larynx, SCPL with CHEP seems to be feasible with acceptable local control and toxicity. Complications may occur as in previously non-irradiated patients. These complications must be treated conservatively to avoid altering laryngeal function. PMID- 20021682 TI - From staff-mix to skill-mix and beyond: towards a systemic approach to health workforce management. AB - Throughout the world, countries are experiencing shortages of health care workers. Policy-makers and system managers have developed a range of methods and initiatives to optimise the available workforce and achieve the right number and mix of personnel needed to provide high-quality care. Our literature review found that such initiatives often focus more on staff types than on staff members' skills and the effective use of those skills. Our review describes evidence about the benefits and pitfalls of current approaches to human resources optimisation in health care. We conclude that in order to use human resources most effectively, health care organisations must consider a more systemic approach- one that accounts for factors beyond narrowly defined human resources management practices and includes organisational and institutional conditions. PMID- 20021684 TI - Chronic kidney-disease screening service quality: questionnaire survey research evidence from Taichung City. AB - BACKGROUND: Chronic kidney disease (CKD) is a serious public health problem in Taiwan and the world. The most effective, affordable treatments involve early prevention/detection/intervention, requiring screening. Successfully implementing CKD programs requires good patient participation, affected by patient perceptions of screening service quality. Service quality improvements can help make such programs more successful. Thus, good tools for assessing service quality perceptions are important. AIM: to investigate using a modified SERVQUAL questionnaire in assessing patient expectations, perceptions, and loyalty towards kidney disease screening service quality. METHOD: 1595 kidney disease screening program patients in Taichung City were requested to complete and return a modified kidney disease screening SERVQUAL questionnaire. 1187 returned them. Incomplete ones (102) were culled and 1085 were chosen as effective for use. Paired t-tests, correlation tests, ANOVA, LSD test, and factor analysis identified the characteristics and factors of service quality. The paired t-test tested expectation score and perception score gaps. A structural equation modeling system examined satisfaction-based components' relationships. RESULTS: The effective response rate was 91.4%. Several methods verified validity. Cronbach's alpha on internal reliability was above 0.902. On patient satisfaction, expectation scores are high: 6.50 (0.82), but perception scores are significantly lower 6.14 (1.02). Older patients' perception scores are lower than younger patients'. Expectation and perception scores for patients with different types of jobs are significantly different. Patients higher on education have lower scores for expectation (r = -0.09) and perception (r = -0.26). Factor analysis identified three factors in the 22 item SERVQUAL form, which account for 80.8% of the total variance for the expectation scores and 86.9% of the total variance for the satisfaction scores. Expectation and perception score gaps in all 22 items are significant. The goodness-of-fit summary of the SEM results indicates that expectations and perceptions are positively correlated, perceptions and loyalty are positively correlated, but expectations and loyalty are not positively correlated. CONCLUSIONS: The results of this research suggest that the SERVQUAL instrument is a useful measurement tool in assessing and monitoring service quality in kidney disease screening services, enabling the staff to identify where service improvements are needed from the patients' perspectives. PMID- 20021683 TI - Baseline hospital performance and the impact of medical emergency teams: modelling vs. conventional subgroup analysis. AB - BACKGROUND: To compare two approaches to the statistical analysis of the relationship between the baseline incidence of adverse events and the effect of medical emergency teams (METs). METHODS: Using data from a cluster randomized controlled trial (the MERIT study), we analysed the relationship between the baseline incidence of adverse events and its change from baseline to the MET activation phase using quadratic modelling techniques. We compared the findings with those obtained with conventional subgroup analysis. RESULTS: Using linear and quadratic modelling techniques, we found that each unit increase in the baseline incidence of adverse events in MET hospitals was associated with a 0.59 unit subsequent reduction in adverse events (95%CI: 0.33 to 0.86) after MET implementation and activation. This applied to cardiac arrests (0.74; 95%CI: 0.52 to 0.95), unplanned ICU admissions (0.56; 95%CI: 0.26 to 0.85) and unexpected deaths (0.68; 95%CI: 0.45 to 0.90). Control hospitals showed a similar reduction only for cardiac arrests (0.95; 95%CI: 0.56 to 1.32). Comparison using conventional subgroup analysis, on the other hand, detected no significant difference between MET and control hospitals. CONCLUSIONS: Our study showed that, in the MERIT study, when there was dependence of treatment effect on baseline performance, an approach based on regression modelling helped illustrate the nature and magnitude of such dependence while sub-group analysis did not. The ability to assess the nature and magnitude of such dependence may have policy implications. Regression technique may thus prove useful in analysing data when there is a conditional treatment effect. PMID- 20021685 TI - Murine mesenchymal progenitor cells from different tissues differentiated via mesenchymal microspheres into the mesodermal direction. AB - BACKGROUND: Because specific marker molecules for phenotypical identification of mesenchymal stem and progenitor cells are missing, the assessment of the in vitro differentiation capacity is a prerequisite to characterize these cells. However, classical differentiation protocols are often cell-consuming and time intensive. Therefore, the establishment of novel strategies for differentiation is one topic of current efforts in stem cell biology. The goal of this study was to demonstrate the practicability of a new differentiation test using plastic adherent cell isolates from different tissues. RESULTS: We introduced the mesenchymal microsphere method as a feasible time- and cell saving screening method to analyse multilineage differentiation properties of adult progenitor cells in a three-dimensional system. For this purpose we isolated, characterized and analyzed new sources of adult murine mesenchymal progenitor cells from perirenal adipose tissue and mediastinal stromal tissue in comparison to bone marrow progenitor cells. The proliferation capacity of the cells was demonstrated by determination of the daily doubling index. Although the flow cytometry analysis of undifferentiated cells revealed differences in the expression of CD marker molecules, all isolates have the capacity for multilineage differentiation following the mesenchymal microsphere protocol as well as the classical "micro mass body" protocol for chondrogenic and the monolayer cultivation protocol for osteogenic and adipogenic differentiation. Differentiation was characterized using histochemical and immunhistochemical staining as well as RT-PCR. CONCLUSIONS: We were able to show that the mesenchymal microsphere method is an efficient test system for chondro-, osteo- and adipogenic differentiation of adult progenitor cells. The advantage of this system in comparison to classical protocols is that approximately 7 times lower cell numbers are necessary. Since classical culture procedures are time intensive because high cell numbers have to be obtained, the new differentiation method may also save cells and time in future clinical applications using human mesenchymal stromal cells. PMID- 20021686 TI - Virulence potential of five major pathogenicity islands (SPI-1 to SPI-5) of Salmonella enterica serovar Enteritidis for chickens. AB - BACKGROUND: Salmonella is a highly successful parasite of reptiles, birds and mammals. Its ability to infect and colonise such a broad range of hosts coincided with the introduction of new genetic determinants, among them 5 major pathogenicity islands (SPI1-5), into the Salmonella genome. However, only limited information is available on how each of these pathogenicity islands influences the ability of Salmonella to infect chickens. In this study, we therefore constructed Salmonella Enteritidis mutants with each SPI deleted separately, with single individual SPIs (i.e. with the remaining four deleted) and a mutant with all 5 SPIs deleted, and assessed their virulence in one-day-old chickens, together with the innate immune response of this host. RESULTS: The mutant lacking all 5 major SPIs was still capable of colonising the caecum while colonisation of the liver and spleen was dependent on the presence of both SPI-1 and SPI-2. In contrast, the absence of SPI-3, SPI-4 or SPI-5 individually did not influence virulence of S. Enteritidis for chickens, but collectively they contributed to the colonisation of the spleen. Proinflammatory signalling and heterophil infiltration was dependent on intact SPI-1 only and not on other SPIs. CONCLUSIONS: SPI-1 and SPI-2 are the two most important pathogenicity islands of Salmonella Enteritidis required for the colonisation of systemic sites in chickens. PMID- 20021687 TI - Identification of ATP binding residues of a protein from its primary sequence. AB - BACKGROUND: One of the major challenges in post-genomic era is to provide functional annotations for large number of proteins arising from genome sequencing projects. The function of many proteins depends on their interaction with small molecules or ligands. ATP is one such important ligand that plays critical role as a coenzyme in the functionality of many proteins. There is a need to develop method for identifying ATP interacting residues in a ATP binding proteins (ABPs), in order to understand mechanism of protein-ligands interaction. RESULTS: We have compared the amino acid composition of ATP interacting and non interacting regions of proteins and observed that certain residues are preferred for interaction with ATP. This study describes few models that have been developed for identifying ATP interacting residues in a protein. All these models were trained and tested on 168 non-redundant ABPs chains. First we have developed a Support Vector Machine (SVM) based model using primary sequence of proteins and obtained maximum MCC 0.33 with accuracy of 66.25%. Secondly, another SVM based model was developed using position specific scoring matrix (PSSM) generated by PSI-BLAST. The performance of this model was improved significantly (MCC 0.5) from the previous one, where only the primary sequence of the proteins were used. CONCLUSION: This study demonstrates that it is possible to predict 'ATP interacting residues' in a protein with moderate accuracy using its sequence. The evolutionary information is important for the identification of 'ATP interacting residues', as it provides more information compared to the primary sequence. This method will be useful for researchers studying ATP-binding proteins. Based on this study, a web server has been developed for predicting 'ATP interacting residues' in a protein http://www.imtech.res.in/raghava/atpint/. PMID- 20021688 TI - Is the use of videotape recording superior to verbal feedback alone in the teaching of clinical skills? AB - BACKGROUND: In recent times, medical schools have committed to developing good communication and history taking skills in students. However, there remains an unresolved question as to which constitutes the best educational method. Our study aims to investigate whether the use of videotape recording is superior to verbal feedback alone in the teaching of clinical skills and the role of student self-assessment on history taking and communication skills. METHODS: A randomized controlled trial was designed. The study was conducted with 52 of the Dokuz Eylul University Faculty of Medicine second year students. All students' performances of communication and history taking skills were assessed twice. Between these assessments, the study group had received both verbal and visual feedback by watching their video recordings on patient interview; the control group received only verbal feedback from the teacher. RESULTS: Although the self-assessment of the students did not change significantly, assessors' ratings increased significantly for videotaped interviews at the second time. CONCLUSIONS: Feedback based on videotaped interviews is superior to the feedback given solely based on the observation of assessors. PMID- 20021689 TI - Association of a functional microsatellite within intron 1 of the BMP5 gene with susceptibility to osteoarthritis. AB - BACKGROUND: In a previous study carried out by our group, the genotyping of 36 microsatellite markers from within a narrow interval of chromosome 6p12.3-q13 generated evidence for linkage and for association to female hip osteoarthritis (OA), with the most compelling association found for a marker within intron 1 of the bone morphogenetic protein 5 gene (BMP5). In this study, we aimed to further categorize the association of variants within intron 1 of BMP5 with OA through an expanded genetic association study of the intron and subsequent functional analysis of associated polymorphisms. METHODS: We genotyped 18 common polymorphisms including 8 microsatellites and 9 single nucleotide polymorphisms (SNPs) and 1 insertion/deletion (INDEL) from within highly conserved regions between human and mouse within intron 1 of BMP5. These markers were then tested for association to OA by a two-stage approach in which the polymorphisms were initially genotyped in a case-control cohort comprising 361 individuals with associated polymorphisms (P < or = 0.05) then genotyped in a second case-control cohort comprising 1185 individuals. RESULTS: Two BMP5 intron 1 polymorphisms demonstrated association in the combined case-control cohort of 1546 individuals (765 cases and 781 controls): microsatellite D6S1276 (P = 0.018) and SNP rs921126 (P = 0.013). Functional analyses in osteoblastic, chondrocytic, and adipocytic cell lines indicated that allelic variants of D6S1276 have significant effects on the transcriptional activity of the BMP5 promoter in vitro. CONCLUSION: Variability in gene expression of BMP5 may be an important contributor to OA genetic susceptibility. PMID- 20021690 TI - Physical activity as a preventive measure against overweight, obesity, infections, allergies and cardiovascular disease risk factors in adolescents: AFINOS Study protocol. AB - BACKGROUND: Prior studies addressing the impacts of regular physical activity or sedentary habits on the immune system have been conducted in adults and laboratory settings. Thus, it is practically unknown how a healthy active lifestyle could affect low-grade inflammation processes, infections or allergies in young persons. The AFINOS Study was designed to determine the relationship between the regular physical activity levels of adolescents and overweight, infection, and allergies along with the presence of metabolic and immunological biomarkers of a deteriorated health status. A further objective of the AFINOS Study is to assess the health status and lifestyle habits of an adolescent population in an effort to identify any protective factors that could be used as preventive measures, since many chronic diseases and their associated co morbidities often persist from adolescence into adulthood. METHODS/DESIGN: This study was conducted as three separate sub-studies in three different populations as follows: (a) Study 1 was performed on a population sample of adolescents; (b) Study 2 on the adolescents' parents; and (c) Study 3 on a subset of the adolescents from Study 1. Study 1 assessed health and lifestyle indicators through a questionnaire administered to a representative sample of adolescents from the Madrid Region (n = 2400) aged 13 to 16 years. In Study 2, the parents of the teenagers participating in Study 1 were required to fill out a questionnaire. Finally in Study 3, body composition, physical activity, health-related physical fitness, and blood measurements were determined in a subset (n = 200) of the individuals included in Study 1. DISCUSSION: This paper describes the rationale, design, and methodologies used in the AFINOS Study. This multidisciplinary, multicenter study seeks to evaluate several aspects of existing relationships between routine physical activity/sedentary behaviour and several health status markers, specifically those related to the immune system. The results of this cross-sectional study will serve for comparisons with the available data obtained in laboratory settings and in adults. In addition, knowledge regarding the health status and lifestyle habits of Spanish adolescents and their parents will be useful for designing preventive measures. PMID- 20021691 TI - Investigation of a community outbreak of typhoid fever associated with drinking water. AB - BACKGROUND: This report is about the investigation of an outbreak of typhoid fever claimed three human lives and left more than 300 people suffered within one week. The aim of this report is to draw the attention of global health community towards the areas that are still far from basic human essentialities. METHODS: A total of 250 suspected cases of typhoid fever were interviewed, out of which 100 were selected for sample collection on the basis of criteria included temperature > 38 degrees C since the onset of outbreak, abdominal discomfort, diarrhea, vomiting and weakness. Food and water samples were also collected and analyzed microbiologically. RESULTS: Inhabitants of village lived in poor and unhygienic conditions with no proper water supply or sewage disposal facilities and other basic necessities of life. They consumed water from a nearby well which was the only available source of drinking water. Epidemiological evidences revealed the gross contamination of well with dead and decaying animal bodies, their fecal material and garbage. Microbiological analysis of household and well water samples revealed the presence of heavy bacterial load with an average total aerobic count 106-109 CFU/ml. A number of Gram positive and Gram negative bacteria including Escherichia coli, Klebsiella, Bacillus species, Staphylococcus species, Enterobacter species, and Pseudomonas aeruginosa were isolated. Lab investigations confirmed the presence of multidrug resistant strain of Salmonella enterica serovar Typhi in 100% well water, 65% household water samples and 2% food items. 22% of clinical stool samples were tested positive with Salmonella enterica serover Typhi CONCLUSIONS: This study indicated the possible involvement of well water in outbreaks. In order to avoid such outbreaks in future, we contacted the local health authorities and urged them to immediately make arrangements for safe drinking water supply. PMID- 20021692 TI - ADHD candidate gene (DRD4 exon III) affects inhibitory control in a healthy sample. AB - BACKGROUND: Dopamine is believed to be a key neurotransmitter in the development of attention-deficit/hyperactivity disorder (ADHD). Several recent studies point to an association of the dopamine D4 receptor (DRD4) gene and this condition. More specifically, the 7 repeat variant of a variable number of tandem repeats (VNTR) polymorphism in exon III of this gene is suggested to bear a higher risk for ADHD. In the present study, we investigated the role of this polymorphism in the modulation of neurophysiological correlates of response inhibition (Go/Nogo task) in a healthy, high-functioning sample. RESULTS: Homozygous 7 repeat carriers showed a tendency for more accurate behavior in the Go/Nogo task compared to homozygous 4 repeat carriers. Moreover, 7 repeat carriers presented an increased nogo-related theta band response together with a reduced go-related beta decrease. CONCLUSIONS: These data point to improved cognitive functions and prefrontal control in the 7 repeat carriers, probably due to the D4 receptor's modulatory role in prefrontal areas. The results are discussed with respect to previous behavioral data on this polymorphism and animal studies on the impact of the D4 receptor on cognitive functions. PMID- 20021693 TI - Phylogeography of the common vampire bat (Desmodus rotundus): marked population structure, Neotropical Pleistocene vicariance and incongruence between nuclear and mtDNA markers. AB - BACKGROUND: The common vampire bat Desmodus rotundus is an excellent model organism for studying ecological vicariance in the Neotropics due to its broad geographic range and its preference for forested areas as roosting sites. With the objective of testing for Pleistocene ecological vicariance, we sequenced a mitocondrial DNA (mtDNA) marker and two nuclear markers (RAG2 and DRB) to try to understand how Pleistocene glaciations affected the distribution of intraspecific lineages in this bat. RESULTS: Five reciprocally monophyletic clades were evident in the mitochondrial gene tree, and in most cases with high bootstrap support: Central America (CA), Amazon and Cerrado (AMC), Pantanal (PAN), Northern Atlantic Forest (NAF) and Southern Atlantic Forest (SAF). The Atlantic forest clades formed a monophyletic clade with high bootstrap support, creating an east/west division for this species in South America. On the one hand, all coalescent and non-coalescent estimates point to a Pleistocene time of divergence between the clades. On the other hand, the nuclear markers showed extensive sharing of haplotypes between distant localities, a result compatible with male-biased gene flow. In order to test if the disparity between the mitochondrial and nuclear markers was due to the difference in mutation rate and effective size, we performed a coalescent simulation to examine the feasibility that, given the time of separation between the observed lineages, even with a gene flow rate close to zero, there would not be reciprocal monophyly for a neutral nuclear marker. We used the observed values of theta and an estimated mutation rate for the nuclear marker gene to perform 1000 iterations of the simulation. The results of this simulation were inconclusive: the number of iterations with and without reciprocal monophyly of one or more clades are similar. CONCLUSIONS: We therefore conclude that the pattern exhibited by the common vampire bat, with marked geographical structure for a mitochondrial marker and no phylogeographic structure for nuclear markers is compatible with a historical scenario of complete isolation of refuge-like populations during the Pleistocene. The results on demographic history on this species is compatible with the Carnaval-Moritz model of Pleistocene vicariance, with demographic expansions in the southern Atlantic forest. PMID- 20021694 TI - TLK1B promotes repair of DSBs via its interaction with Rad9 and Asf1. AB - BACKGROUND: The Tousled-like kinases are involved in chromatin assembly, DNA repair, transcription, and chromosome segregation. Previous evidence indicated that TLK1B can promote repair of plasmids with cohesive ends in vitro, but it was inferred that the mechanism was indirect and via chromatin assembly, mediated by its interaction with the chromatin assembly factor Asf1. We recently identified Rad9 as a substrate of TLK1B, and we presented evidence that the TLK1B-Rad9 interaction plays some role in DSB repair. Hence the relative contribution of Asf1 and Rad9 to the protective effect of TLK1B in DSBs repair is not known. Using an adeno-HO-mediated cleavage system in MM3MG cells, we previously showed that overexpression of either TLK1B or a kinase-dead protein (KD) promoted repair and the assembly of Rad9 in proximity of the DSB at early time points post infection. This established that it is a chaperone activity of TLK1B and not directly the kinase activity that promotes recruitment of 9-1-1 to the DSB. However, the phosphorylation of Rad9(S328) by TLK1B appeared important for mediating a cell cycle checkpoint, and thus, this phosphorylation of Rad9 may have other effects on 9-1-1 functionality. RESULTS: Here we present direct evidence that TLK1B can promote repair of linearized plasmids with incompatible ends that require processing prior to ligation. Immunodepletion of Rad9 indicated that Rad9 was important for processing the ends preceding ligation, suggesting that the interaction of TLK1B with Rad9 is a key mediator for this type of repair. Ligation of incompatible ends also required DNA-PK, as addition of wortmannin or immunodepletion of Ku70 abrogated ligation. Depletion of Ku70 prevented the ligation of the plasmid but did not affect stimulation of the fill in of the ends by added TLK1B, which was attributed to Rad9. From experiments with the HO-cleavage system, we now show that Rad17, a subunit of the "clamp loader", associates normally with the DSB in KD-overexpressing cells. However, the subsequent release of Rad17 and Rad9 upon repair of the DSB was significantly slower in these cells compared to controls or cells expressing wt-TLK1B. CONCLUSIONS: TLKs play important roles in DNA repair, not only by modulation of chromatin assembly via Asf1, but also by a more direct function in processing the ends of a DSB via interaction with Rad9. Inhibition of Rad9 phosphorylation in KD overexpressing cells may have consequences in signaling completion of the repair and cell cycle re-entry, and could explain a loss of viability from DSBs in these cells. PMID- 20021695 TI - Genome-wide profiling of populus small RNAs. AB - BACKGROUND: Short RNAs, and in particular microRNAs, are important regulators of gene expression both within defined regulatory pathways and at the epigenetic scale. We investigated the short RNA (sRNA) population (18-24 nt) of the transcriptome of green leaves from the sequenced Populus trichocarpa using a concatenation strategy in combination with 454 sequencing. RESULTS: The most abundant size class of sRNAs were 24 nt. Long Terminal Repeats were particularly associated with 24 nt sRNAs. Additionally, some repetitive elements were associated with 22 nt sRNAs. We identified an sRNA hot-spot on chromosome 19, overlapping a region containing both the proposed sex-determining locus and a major cluster of NBS-LRR genes. A number of phased siRNA loci were identified, a subset of which are predicted to target PPR and NBS-LRR disease resistance genes, classes of genes that have been significantly expanded in Populus. Additional loci enriched for sRNA production were identified and characterised. We identified 15 novel predicted microRNAs (miRNAs), including miRNA*sequences, and identified a novel locus that may encode a dual miRNA or a miRNA and short interfering RNAs (siRNAs). CONCLUSIONS: The short RNA population of P. trichocarpa is at least as complex as that of Arabidopsis thaliana. We provide a first genome-wide view of short RNA production for P. trichocarpa and identify new, non-conserved miRNAs. PMID- 20021696 TI - The longitudinal changes of BOLD response and cerebral hemodynamics from acute to subacute stroke. A fMRI and TCD study. AB - BACKGROUND: By mapping the dynamics of brain reorganization, functional magnetic resonance imaging MRI (fMRI) has allowed for significant progress in understanding cerebral plasticity phenomena after a stroke. However, cerebro vascular diseases can affect blood oxygen level dependent (BOLD) signal. Cerebral autoregulation is a primary function of cerebral hemodynamics, which allows to maintain a relatively constant blood flow despite changes in arterial blood pressure and perfusion pressure. Cerebral autoregulation is reported to become less effective in the early phases post-stroke. This study investigated whether any impairment of cerebral hemodynamics that occurs during the acute and the subacute phases of ischemic stroke is related to changes in BOLD response. We enrolled six aphasic patients affected by acute stroke. All patients underwent a Transcranial Doppler to assess cerebral autoregulation (Mx index) and fMRI to evaluate the amplitude and the peak latency (time to peak-TTP) of BOLD response in the acute (i.e., within four days of stroke occurrence) and the subacute (i.e., between five and twelve days after stroke onset) stroke phases. RESULTS: As patients advanced from the acute to subacute stroke phase, the affected hemisphere presented a BOLD TTP increase (p = 0.04) and a deterioration of cerebral autoregulation (Mx index increase, p = 0.046). A similar but not significant trend was observed also in the unaffected hemisphere. When the two hemispheres were grouped together, BOLD TTP delay was significantly related to worsening cerebral autoregulation (Mx index increase) (Spearman's rho = 0.734; p = 0.01). CONCLUSIONS: The hemodynamic response function subtending BOLD signal may present a delay in peak latency that arises as patients advance from the acute to the subacute stroke phase. This delay is related to the deterioration of cerebral hemodynamics. These findings suggest that remodeling the fMRI hemodynamic response function in the different phases of stroke may optimize the detection of BOLD signal changes. PMID- 20021697 TI - Comparison of linkage disequilibrium and haplotype diversity on macro- and microchromosomes in chicken. AB - BACKGROUND: The chicken (Gallus gallus), like most avian species, has a very distinct karyotype consisting of many micro- and a few macrochromosomes. While it is known that recombination frequencies are much higher for micro- as compared to macrochromosomes, there is limited information on differences in linkage disequilibrium (LD) and haplotype diversity between these two classes of chromosomes. In this study, LD and haplotype diversity were systematically characterized in 371 birds from eight chicken populations (commercial lines, fancy breeds, and red jungle fowl) across macro- and microchromosomes. To this end we sampled four regions of approximately 1 cM each on macrochromosomes (GGA1 and GGA2), and four 1.5 -2 cM regions on microchromosomes (GGA26 and GGA27) at a high density of 1 SNP every 2 kb (total of 889 SNPs). RESULTS: At a similar physical distance, LD, haplotype homozygosity, haploblock structure, and haplotype sharing were all lower for the micro- as compared to the macrochromosomes. These differences were consistent across populations. Heterozygosity, genetic differentiation, and derived allele frequencies were also higher for the microchromosomes. Differences in LD, haplotype variation, and haplotype sharing between populations were largely in line with known demographic history of the commercial chicken. Despite very low levels of LD, as measured by r2 for most populations, some haploblock structure was observed, particularly in the macrochromosomes, but the haploblock sizes were typically less than 10 kb. CONCLUSION: Differences in LD between micro- and macrochromosomes were almost completely explained by differences in recombination rate. Differences in haplotype diversity and haplotype sharing between micro- and macrochromosomes were explained by differences in recombination rate and genotype variation. Haploblock structure was consistent with demography of the chicken populations, and differences in recombination rates between micro- and macrochromosomes. The limited haploblock structure and LD suggests that future whole-genome marker assays will need 100+K SNPs to exploit haplotype information. Interpretation and transferability of genetic parameters will need to take into account the size of chromosomes in chicken, and, since most birds have microchromosomes, in other avian species as well. PMID- 20021698 TI - Serum macrophage migration inhibitory factor reflects adrenal function in the hypothalamo-pituitary-adrenal axis of septic patients: an observational study. AB - BACKGROUND: The hypothalamo-pituitary-adrenal (HPA) axis modulates the inflammatory response during sepsis. Macrophage migration inhibitory factor (MIF), which counteracts the anti-inflammatory activity of glucocorticoid (GC), is one of the mediators of the development of inflammation. An inflammatory imbalance involving GC and MIF might be the cause or result of adrenal insufficiency. Our objective was to clarify the relationship between serum MIF and adrenal function in the HPA axis of sepsis patients using the adrenocorticotropic hormone (ACTH) stimulation test. METHODS: An observational study was performed in a university intensive care unit over a two-year period. Of 64 consecutive sepsis patients, 41 were enrolled. The enrolled patients underwent an ACTH stimulation test within 24 h of the diagnosis of severe sepsis or septic shock. Clinical and laboratory parameters, including serum MIF and cortisol, were measured. RESULTS: Based on their responses to the ACTH stimulation test, the patients were divided into a normal adrenal response (NAR) group (n = 22) and an adrenal insufficiency (AI) group (n = 19). The AI group had significantly more septic shock patients and higher prothrombin time ratios, serum MIF, and baseline cortisol than did the NAR group (P < 0.05). Serum MIF correlated significantly with the SOFA (Sequential Organ Failure Assessment) score, prothrombin time ratio, and delta max cortisol, which is maximum increment of serum cortisol concentration after ACTH stimulation test (rs = 0.414, 0.355, and -0.49, respectively, P < 0.05). Serum MIF also correlated significantly with the delta max cortisol/albumin ratio (rs = -0.501, P = 0.001). Receiver operating characteristic curve analysis identified the threshold serum MIF concentration (19.5 ng/mL, P = 0.01) that segregated patients into the NAR and AI groups. CONCLUSIONS: The inverse correlation between serum MIF and delta max cortisol or the delta max cortisol/albumin ratio suggests that high serum MIF reflects an insufficient adrenal response in the HPA axis. Serum MIF could be a valuable clinical marker of adrenal insufficiency in sepsis patients. PMID- 20021700 TI - High-starch diets induce precocious adipogenic gene network up-regulation in longissimus lumborum of early-weaned Angus cattle. AB - Adipocyte differentiation is probably controlled by transcriptional and post transcriptional regulation. Longissimus lumborum from Angus steers (aged 155 d; seven animals per diet) fed high-starch or low-starch diets for 112 d (growing phase) followed by a common high-starch diet for an additional 112 d (finishing phase) was biopsied at 0, 56, 112 and 224 d for transcript profiling via quantitative PCR of twenty genes associated with adipogenesis and energy metabolism. At 56 d steers fed high starch had greater expression of PPARgamma as well as the lipogenic enzymes ATP citrate lyase (ACLY), glucose-6-phosphate dehydrogenase (G6PD), fatty acid synthase (FASN), fatty acid binding protein 4 (FABP4), stearoyl-CoA desaturase (SCD), glycerol-3-phosphate acyltransferase, mitochondrial (GPAM), and diacylglycerol O-acyltransferase homologue 2 (DGAT2), and the adipokine adiponectin (ADIPOQ). Expression of insulin-induced gene 1 (INSIG1) was also greater with high starch at 56 d. Steers fed low starch experienced a marked increase in FASN, FABP4, SCD, DGAT2 and thyroid hormone responsive (SPOT14 homologue, rat) (THRSP) between 56 and 112 d of feeding. A greater expression of the transcription factors sterol regulatory element-binding transcription factor 1 (SREBF1) and MLX interacting protein-like (MLXIPL) was observed at 224 d in steers fed high starch, suggesting a nutritional imprinting effect. Carryover effects of low starch feeding were discerned by greater expression at 224 d of THRSP, FABP4, SCD and DGAT2. These steers also had greater PPARgamma at 224 d. Despite these responses, low starch led to greater expression at 224 d of nuclear receptor subfamily 2, group F, member 2 (NR2F2), a known repressor of rodent adipocyte differentiation through its negative effects on PPARgamma, ADIPOQ and FABP4. Results suggested that early exposure to high starch induced precocious intramuscular adipocyte proliferation and metabolic imprinting of lipogenic transcription regulators. Low starch might have blunted the PPARgamma-driven adipogenic response through up-regulation of NR2F2 but the endogenous ligand for this nuclear receptor remains unknown. PMID- 20021699 TI - Intrinsic chemoresistance to gemcitabine is associated with constitutive and laminin-induced phosphorylation of FAK in pancreatic cancer cell lines. AB - BACKGROUND: One of the major reasons for poor prognosis of pancreatic cancer is its high resistance to currently available chemotherapeutic agents. In recent years, focal adhesion kinase (FAK), a central molecule in extracellular matrix (ECM)/integrin-mediated signaling, has been thought to be a key determinant of chemoresistance in cancer cells. In this study, we aimed to determine the roles of FAK phosphorylation in the intrinsic chemoresistance of pancreatic cancer cell lines. RESULTS: Our results showed that, the level of constitutive phosphorylation of FAK at Tyr397 correlated with the extent of intrinsic resistance to Gemcitabine (Gem) in four pancreatic cancer cell lines. Moreover, in Panc-1 cells, which had high expression of pFAK, specific inhibition of constitutive FAK phosphorylation by either RNAi or FRNK overexpression decreased the phosphorylation of Akt, reduced the levels of survivin expression and Bad phosphorylation at Ser136 and increased Gem-induced cytotoxicity and apoptosis. However, in AsPC-1 cells with a low level of pFAK, neither FAK RNAi nor FRNK overexpression affected Gem-induced cell apoptosis. We further found that laminin (LN) induced FAK and Akt phosphorylation in a time-dependent manner, increased the levels of survivin and pBad (pS136) and decreased Gem-induced cytotoxicity and apoptosis in AsPC-1 cells; Specific inhibition of LN-induced FAK phosphorylation by either FAK RNAi or FRNK overexpression suppressed the effects of LN on AsPC-1 cells. Moreover, inhibition of constitutive FAK phosphorylation in Panc-1 cells and LN-induced FAK phosphorylation in AsPC-1 cells by a novel and more specific FAK phosphorylation inhibitor PF-573,228 showed similar results with those of FAK phosphorylation inhibition by FAK RNAi or FRNK overexpression. CONCLUSIONS: In conclusion, our research demonstrates for the first time that both constitutive and LN-induced FAK phosphorylation contribute to increased intrinsic chemoresistance to Gem in pancreatic cancer cell lines and these effects are partly due to the regulation of Akt and Bad phosphorylation and survivin expression. Development of selective FAK phosphorylation inhibitors may be a promising way to enhance chemosensitivity in pancreatic cancer. PMID- 20021701 TI - Predicting in vivo starch digestibility coefficients in newly weaned piglets from in vitro assessment of diets using multivariate analysis. AB - The study was based on correlating a dataset of in vivo mean starch digestibility coefficients obtained in the immediate post-weaning phase of piglets with a range of dietary in vitro variables. The paper presents a model that predicts (R2 0.71) in vivo average starch digestibility coefficients in the 0.5 small-intestinal region of newly weaned piglets fed cereal-based diets using seven in vitro variables describing starch properties that are fundamentally associated with the quality of feed materials, i.e. hydration, structure and amylolytic digestion. The variables were: Rapid Visco Analyser (RVA; measures the viscosity of materials when sheared under defined hydration and temperature regimens); RVA end viscosity; RVA (gelatinisation) peak viscosity; DeltaH (gelatinisation enthalpy that provides an estimate of helical order or degree of crystallinity in starch); water solubility index (WSI; that denotes the amount of soluble polysaccharides released from starch granules to the aqueous phase); grain endogenous amylase (concentration of endogenous alpha-amylase in cereals, assessed by pasting cereal flours in 25 g of AgNO3, an amylase inhibitor v. water using RVA). PMID- 20021702 TI - Physiological concentrations of dietary polyphenols regulate vascular endothelial cell expression of genes important in cardiovascular health. AB - Previous cell culture-based studies have shown potential health beneficial effects on gene expression of dietary polyphenols, including those found in red wine and green tea. However, these studies have tended to use higher concentrations (2-100 microm) than those observed in blood (0.1-1 microm) after consuming polyphenol-rich foods or beverages. The present study investigated effects of physiological concentrations of different classes of dietary polyphenol on the expression of genes important in cardiovascular health (endothelial NO synthase (eNOS), endothelin-1 (ET-1) and vascular endothelial growth factor (VEGF)) by cultured vascular endothelial cells (human umbilical vein endothelial cells) in the absence or presence of H2O2. Resveratrol and quercetin (0.1-1 microm) increased eNOS and VEGF mRNA expression particularly in the absence of H2O2 (50 microm) and decreased H2O2-induced ET-1 mRNA expression (P < 0.001 for polyphenol x H2O2 interactions). Similarly, resveratrol and quercetin decreased endothelin secretion into the media, blocking the stimulatory effect of 50 microm-H2O2 (P < 0.001 for polyphenol x H2O2 interaction). Of the nine other polyphenols tested, only epigallocatechin gallate had similar effects on both the eNOS and ET-1 mRNA expression, but to a lesser extent than resveratrol at an equimolar concentration (0.1 microm). The observed effects on gene expression would be expected to result in vasodilation and thereby reduced blood pressure. Since only three of the eleven polyphenols tested had biological activity, it is unclear whether particular structures are important or whether the effects might relate to the relatively high antioxidant capacities of the three active polyphenols. PMID- 20021703 TI - The effect of a multispecies probiotic on the composition of the faecal microbiota and bowel habits in chronic obstructive pulmonary disease patients treated with antibiotics. AB - Short-term antibiotic treatment profoundly affects the intestinal microbiota, which may lead to sustained changes in microbiota composition. Probiotics may restore such a disturbance. The objective of the present study was to investigate the effect of a multispecies probiotic on the faecal microbiota during and after antibiotic intake in patients with a history of frequent antibiotic use. In this randomised, placebo-controlled, double-blind study, thirty chronic obstructive pulmonary disease (COPD) patients treated with antibiotics for a respiratory tract infection received 5 g of a multispecies probiotic or placebo twice daily for 2 weeks. Faecal samples were collected at 0, 7, 14 and 63 d. Changes in the composition of the dominant faecal microbiota were determined by PCR-denaturing gradient gel electrophoresis (DGGE). Changes in bacterial subgroups were determined by quantitative PCR and culture. Bowel movements were scored daily according to the Bristol stool form scale. During and after antibiotic treatment, DGGE-based similarity indices (SI) were high ( >/= 84 %) and band richness was relatively low, both remaining stable over time. No difference in SI was observed between patients with and without diarrhoea-like bowel movements. The multispecies probiotic had a modest effect on the bacterial subgroups. Nevertheless, it affected neither the composition of the dominant faecal microbiota nor the occurrence of diarrhoea-like bowel movements. The dominant faecal microbiota was not affected by antibiotics in this COPD population, suggesting an existing imbalance of the microbiota, which may also have contributed to the lack of effect by probiotic intake. PMID- 20021704 TI - Starch-entrapped microspheres show a beneficial fermentation profile and decrease in potentially harmful bacteria during in vitro fermentation in faecal microbiota obtained from patients with inflammatory bowel disease. AB - The purpose of this research was to test the hypothesis that starch-entrapped microspheres would produce favourable fermentation profiles and microbial shifts during in vitro fermentation with the faecal microbiota from patients with inflammatory bowel disease (IBD). In vitro fermentation was carried out using a validated, dynamic, computer-controlled model of the human colon (Toegepast Natuurwetenschappelijk Onderzoek gastro-intestinal model-2) after inoculation with pooled faeces from healthy individuals, patients with inactive IBD (Crohn's disease (CD)) or patients with active IBD (ulcerative colitis (UC)). Starch entrapped microspheres fermented more slowly and produced more butyrate than fructo-oligosaccharides (FOS) when fermented with the faecal microbiota from patients with active UC. When fermented with the microbiota from patients with inactive CD, starch-entrapped microspheres also fermented more slowly but produced similar amounts of butyrate compared with FOS. Starch-entrapped microspheres showed a greater ability to maintain a low pH during simulated distal colon conditions compared with FOS. After fermentation with the microbiota from inactive CD patients, starch-entrapped microspheres resulted in lower concentrations of some potentially harmful gut bacteria, included in Bacteroides, Enterococcus, Fusobacterium and Veillonella, compared with FOS. These findings suggest that slow fermenting starch-entrapped microspheres may induce a favourable colonic environment in patients with IBD through high butyrate production, maintenance of low pH in the distal colon and inhibition of the growth of potentially harmful bacteria. PMID- 20021705 TI - Effect of prebiotic fibre supplementation on hepatic gene expression and serum lipids: a dose-response study in JCR:LA-cp rats. AB - Prebiotic fibres have been proposed to promote weight loss and lower serum cholesterol; however, the mechanisms are not fully understood. The aim of the present research was to identify possible mechanisms through which prebiotic fibres improve serum lipids. Lean and obese JCR:La-cp rats aged 8 weeks consumed one of three diets supplemented with 0, 10 or 20 % prebiotic fibre for 10 weeks. Rats were anaesthetised and a fasting blood sample was taken for lipid analysis. Real-time PCR was used to determine gene expression for cholesterol and fatty acid regulatory genes in liver tissue. Liver and caecal digesta cholesterol and TAG content were quantified. Both doses of prebiotic fibre lowered serum cholesterol levels by 24 % in the obese hyperlipidaemic rats (P < 0.05). This change was associated with an increase in caecal digesta as well as an up regulation of genes involved in cholesterol synthesis and bile production. Additionally, there was a 42 % reduction in TAG accumulation in the liver of the obese rats with 10 % prebiotic diet (P < 0.05); however, no change in liver fatty acid synthase (FAS). Prebiotic fibres appear to lower cholesterol levels through increased cholesterol excretion in the form of bile and inhibit the accumulation of TAG in the liver through a mechanism unrelated to FAS. These effects appear to be limited to the obese model and particularly the 10 % dose. The present work is significant as it provides insight into the mechanisms of action for prebiotic fibres on lipid metabolism and furthers the development of dietary treatments for hypercholesterolaemia. PMID- 20021706 TI - Only human: hostile human norms can reduce legitimization of intergroup discrimination by perpetrators of historical atrocities. AB - We investigated the effects of salient shared humanity with a benevolent or hostile human norm on perpetrators of historical atrocities. Our findings suggest that a focus on benevolent superordinate humanity enables perpetrators to legitimize intergroup discrimination and preserve existing negative attitudes towards victims. In Expt 1 (N=135), salient shared humanity with a human norm of benevolence and kindness preserved the perceived legitimacy of intergroup inequality, while exposure to a hostile norm of human nature reduced perceived legitimacy. Expt 2 (N=51) replicated the association between exposure to a hostile human norm and reduced legitimization when perpetrator intentions were unambiguously negative. In contrast, when perpetrator intentions were ambiguous, a hostile human norm had no effect on perceived legitimacy. Our findings qualify previous research, and demonstrate that the effects of emphasizing shared humanity are not equivalent or universally positive for perpetrators and victims. PMID- 20021707 TI - Explaining prosocial intentions: testing causal relationships in the norm activation model. AB - This paper examines factors influencing prosocial intentions. On the basis of the norm activation model (NAM), we propose that four variables influence prosocial intentions or behaviours: (1) personal norms (PN), reflecting feelings of moral obligation to engage in prosocial behaviour, (2) awareness of adverse consequences of not acting prosocially, (3) ascription of responsibility for the negative consequences of not acting prosocially, and (4) perceived control over the problems. We conducted a series of experimental studies to examine how the NAM variables are causally related. As hypothesized, problem awareness, responsibility, and outcome efficacy played an important role in the development of PN and various types of prosocial intentions in the social as well as environmental domain. PMID- 20021708 TI - Children's printed word database: continuities and changes over time in children's early reading vocabulary. AB - In this paper we introduce a comprehensive database of the vocabulary in reading materials used by 5 - 9 year old children in the UK. The database is available on line http://www.essex.ac.uk/psychology/cpwd and allows researchers into early reading development the possibility of rigorous control over critical characteristics of experimental stimuli such as word frequency, regularity and length, frequency of grapheme-phoneme correspondences, orthographic and phonological neighbourhoods etc. The on-line database is also a resource that can be used by practitioners with interests in literacy development and literacy instruction. It can be used to obtain characteristics for a user-generated list of words, or else to generate a list of words according to constraints specified by the user. Here we present an overview of the construction of the database, the materials entered into it, the survey of schools by which we obtained information about the books that were most likely to be used by children in each age group, and the search features available on the database website. We also discuss certain characteristics of the Vocabulary itself and compare these with those reported in an earlier non-representative database reported in Stuart, Dixon, Masterson and Gray (2003). We then present a detailed analysis of the characteristics of Vocabulary in books used in the Reception year, against the background of recent recommendations for change in the early teaching of reading. Finally, we present data showing that the database is indeed already proving a useful resource for both practitioners and researchers. PMID- 20021709 TI - Odour-based context reinstatement effects with indirect measures of memory: the curious case of rosemary. AB - Previous studies examining environmental context-dependent memory (ECDM) effects using indirect measures of memory have produced inconsistent findings. We report three experiments that examined ECDM in an indirect memory paradigm (word fragment completion) using ambient odours as environmental contexts. Expt 1 manipulated the odour present at learning and testing (rosemary or lemon) to produce reinstated-context or switched-context conditions. Reinstating rosemary led to a striking ECDM effect, indicating that indirect memory testing can be sensitive to ECDM manipulations. Odour ratings also indicated that rosemary induced a more unpleasant mood in participants than lemon. Expt 2 assessed the influence on indirect retrieval of odour-based mood induction as well as odour distinctiveness, and indicated that rosemary's capacity to promote ECDM effects appears to arise from an additive combination of its unpleasantness-inducing properties and its distinctiveness. Expt 3 partially supported these proposals. Overall, our findings indicate that some odours are capable of producing ECDM effects using indirect testing procedures. Moreover, it appears that it is the inherent proprieties of odours on dimensions such as unpleasantness and distinctiveness that mediate the emergence of ECDM effects, thereby explaining the particular potency of rosemary's mnemonic influence when it is reinstated. PMID- 20021710 TI - Parental choice: what parents want in a son-in-law and a daughter-in-law across 67 pre-industrial societies. AB - Parents are influential over mate choice, and in most human societies they choose spouses for their offspring according to their own preferences. However, surprising little is known about the qualities which make a woman desirable as a daughter-in-law and a man desirable as a son-in-law. Using evidence from 67 societies such traits are identified and three hypotheses are tested: first, the hypothesis is tested that parents desire in an in-law qualities which are beneficial to them and their kin. Second, it is hypothesized that such preferences are contingent upon the sex of the in-law, as traits are weighted differently in a daughter-in-law and in a son-in-law. The third hypothesis tested is that parental preferences vary according to the subsistence type of a given society, as traits are valued differently in agropastoral societies and foraging societies. The evidence presented here provides support for all three hypotheses. PMID- 20021711 TI - Low-dose HCG may improve pregnancy rates and lower OHSS in antagonist cycles: a meta-analysis. AB - Human chorionic gonadotrophin (HCG) may substitute FSH to complete follicular growth in IVF cycles. This may be useful in the prevention of ovarian hyperstimulation syndrome. Relevant studies were identified on Medline. To evaluate outcomes, a meta-analysis of low-dose HCG-supplemented IVF cycles versus non-supplemented ones was performed with data from 435 patients undergoing IVF who were administered low-dose HCG in various agonist and antagonist protocols and from 597 conservatively treated patients who served, as control subjects. Using these published data, a decision analysis evaluated four different management strategies. Effectiveness and economic outcomes were assessed by FSH consumption, clinical pregnancy and incremental cost-effectiveness ratios. Clinical pregnancy and ovarian hyperstimulation were the main outcome measures. Nine trials published in 2002-2007 were included. From the prospective studies, in the gonadotrophin-releasing hormone antagonist group, a trend for significance in clinical pregnancy rate was evident (odds ratio [OR], 1.54; 95% confidence interval [CI], 0.98-2.42). Ovarian hyperstimulation was less significant in the antagonist low-dose HCG protocol compared with the non-supplemented agonist protocol (OR 0.30; 95% CI 0.09-0.96). Less FSH was consumed in the low-dose HCG group but this difference was not statistically significant. Low-dose HCG supplementation may improve pregnancy rates in antagonist protocols. Overall, low dose HCG-supplemented protocols are a cost-effective strategy. PMID- 20021712 TI - Follicular-fluid anti-Mullerian hormone concentrations are predictive of assisted reproduction outcome in PCOS patients. AB - Serum anti-Mullerian hormone (AMH) concentrations constitute a sensitive marker for ovarian ageing. In addition, concentrations of AMH in the follicular fluid constitute a useful marker of embryo implantation in assisted reproduction cycles. The present study measured serum and follicular-fluid AMH concentrations on the day of oocyte retrieval. These data showed that clinical pregnancy rates (25.0, 34.1 and 42.1%, respectively, P < 0.001), embryo implantation rates (24.3, 35.0 and 44.4%, respectively, P < 0.001) and fertilization rates (59.2, 70.9 and 79.5%, respectively, P < 0.001) were markedly different among the low, moderate and high follicular-fluid AMH groups but not among the different serum AMH concentration groups. Follicular-fluid AMH concentrations were negatively correlated with follicular-fluid oestradiol concentrations. The results of this study suggest that follicular-fluid AMH concentration on the day of oocyte retrieval would appear to better reflect the reproductive outcome in PCOS patients undergoing assisted reproduction. PMID- 20021713 TI - Oxidative stress and medical antioxidant treatment in male infertility. AB - Oxidative stress (OS) has been recognized as one of the most important cause of male infertility. Despite the antioxidant activity of seminal plasma, epididymis and spermatozoa, OS damages sperm function and DNA integrity. Since antioxidants suppress the action of reactive oxygen species, these compounds have been used in the medical treatment of male infertility or have been added to the culture medium during sperm separation techniques. Nevertheless, the efficacy of such a treatment has been reported to be very limited. This may relate to: (i) patient selection bias; (ii) late diagnosis of male infertility; (iii) lack of double blind, placebo-controlled clinical trial; and/or (iv) use of end-points that are not good markers of the presence of OS. This review considers the effects of the main antioxidant compounds used in clinical practice. Overall, the data published suggest that no single antioxidant is able to enhance fertilizing capability in infertile men, whereas a combination of them seems to provide a better approach. Taking into account the pros and the cons of antioxidant treatment of male infertility, the potential advantages that it offers cannot be ignored. Therefore, antioxidant therapy should remain in the forefront of preventive medicine, including human reproductive medicine. PMID- 20021714 TI - Identification of diabetes- and obesity-associated proteomic changes in human spermatozoa by difference gel electrophoresis. AB - Difference gel electrophoresis (DIGE) of fluorescently labelled human sperm proteins was used to identify diabetes- and obesity-associated changes of the sperm proteome. Semen samples from type 1 diabetics, non-diabetic obese individuals and a reference group of clinically healthy fertile donors were evaluated in a comparative study. The adaptation of a general protein extraction procedure to the solubilization of proteins from isolated progressively motile human spermatozoa resulted in the detection of approximately 2700 fluorescent protein spots in the DIGE images. Comparison of the patients' sperm proteomes with those of the reference group allowed the identification of 20 spots containing proteins that were present in the sperm lysates at significantly increased or decreased concentrations. In detail, eight of these spots were apparently related to type 1 diabetes while 12 spots were apparently related to obesity. Tryptic digestion of the spot proteins and mass spectrometric analysis of the corresponding peptides identified seven sperm proteins apparently associated with type 1 diabetes and nine sperm proteins apparently associated with obesity, three of which existing in multiple molecular forms. The established proteomic approach is expected to function as a non-invasive experimental tool in the diagnosis of male infertility and in monitoring any fertility-restoring therapy. PMID- 20021715 TI - Chromomycin A3 staining, sperm chromatin structure assay and hyaluronic acid binding assay as predictors for assisted reproductive outcome. AB - Functional sperm tests such as the sperm chromatin structure assay (SCSA), chromomycin A3 staining (CMA(3)) and hyaluronic acid binding assay (HBA) have been suggested as predictive tests of fertility in vitro. This study aimed to define the clinical role of these functional parameters in assisted reproduction in a prospective cohort study. Conventional sperm diagnosis (motility, morphology and concentration) as well as SCSA, CMA(3) and HBA tests were performed on 205 semen samples [74 IVF, 94 ICSI and 37 combined IVF/intracytoplasmic sperm injection (ICSI)]. Main outcome parameters were fertilization rate, clinical pregnancy rate and take-home baby rate. The study showed that each of the three functional sperm tests was related to one or more conventional and one or more functional sperm tests, indicating that spermatozoa from patients with abnormal conventional semen parameters have a higher likelihood for multiple functional abnormalities. Only SCSA and CMA(3) staining were shown to have a limited predictive value when IVF or combined IVF/ICSI was applied. The proposed threshold value of =12 (HADS) and >=13 (BDI). Sensitivity was 81.0% (HADS) and 75.0% (BDI). Specificity was 90.2% for both. CONCLUSIONS: Both HADS and BDI are valid screening instruments for the diagnosis depression in ESRD patients but there is no statistical difference found between both rating scales. PMID- 20021731 TI - The meaning of self-injury and overdosing amongst women fulfilling the diagnostic criteria for 'borderline personality disorder'. AB - OBJECTIVES: To explore the meanings of self-injury and overdosing, and the relationship of each to the other, for women who have fulfilled the diagnostic criteria for borderline personality disorder. DESIGN: Four women with a history of both self-injury and overdosing were recruited through a local psychotherapy service and interviewed following a semi-structured framework. METHODS: The interview transcripts were systematically analysed utilizing interpretative phenomenological analysis. RESULTS: The analysis lead to the construction of three interrelated themes: (a) The context of distress. Examples were inclusive of temporally distal factors such as childhood abuse and loss, and proximal factors ranging from the interpersonal (e.g., conflict) through to the intra personal (e.g., negative thoughts relating to self). (b) The progressive management of distress. An overdose was depicted as a last resort once self injury had become insufficient a means of self-help to manage feelings of desperation and isolation. (c) Ambivalence in relation to death. The clearly stated intention of an overdose was 'to die'. However, both words and (descriptions of) deeds suggested considerable ambivalence in relation to this- overdoses appearing to depict an attempt to resolve unbearable feelings through an unarticulated 'cry for help' rather than suicide completion. CONCLUSIONS: The overall impression gained was of a group of women in a world of problematic relations (to both self and others) struggling to know how best to attract support and understanding. In discussion, links are made to the literature on reflectivity/reflexivity through Fonagy's notion of 'mentalization'. PMID- 20021732 TI - The Swedish version of the Outcome Questionnaire (OQ-45): reliability and factor structure in a substance abuse sample. AB - This study described the psychometric properties of the Swedish version of the Outcome Questionnaire (OQ-45.2) in a substance abuse sample (N=227). The test properties were satisfactory except for somewhat low internal consistency in the social role subscale. The results were similar to those obtained in other countries but systematic international comparisons are still lacking. PMID- 20021733 TI - Transplantation of long-term cultured porcine islets in the rat: prolonged graft survival and recipient growth on reduced immunosuppression. AB - To evaluate whether further improvement in porcine islet xenotransplantation is feasible, a number of questions were addressed. Earlier we showed significant improvement in the nude mouse of the porcine islets by selection through long term culture. Now these islets were tested in the stringent pig-to-rat model. Islets were isolated from adult pigs, cultured for 1.5-3 weeks and transplanted to rats. Possible rejection mechanisms were assessed by interference of the cellular response with cyclosporine A (CsA), blocking macrophages with gadolinium chloride (GdCl), and suppressing the humoral response with cyclophosphamide. Modifications in graft size and condition were analyzed. Untreated control recipients showed primary nonfunction (PNF). CsA treatment could fully overcome PNF and resulted in graft survival from 10 to over 134 days. Rejection was the main cause of function loss. Although rejection could not be prevented by intensifying the induction therapy, increased maintenance immunosuppression effectively blocked rejection, albeit at the expense of toxicity. Blocking the humoral response was ineffective; all grafts showed PNF. In contrast, depletion of macrophages fully prevented PNF. Combination of GdCl and CsA gave no additional effect, and grafts were rejected between 57 and 162 days. Generally, graft survivals were similar to those reported in the literature; however, long term cultured islets required much less maintenance immunosuppression. Cessation of graft function was not always due to rejection; in some cases "islet exhaustion" was found, possibly caused by discrepancy between the graft size and the rapidly growing recipient. Neither the presence of damaged islet tissue in the graft nor the size of the graft exerted any influence on graft survival. On rejection, no real infiltration of the graft was seen; destruction gradually processed from the outside. The good functional capability of the cultured islets was illustrated by disappearance of the clinical symptoms and increase in body weight, which almost doubled in the long-term survivors. PMID- 20021734 TI - Analysis of in vitro and in vivo characteristics of human embryonic stem cell derived neural precursors. AB - During the last decade, much progress has been made in developing protocols for the differentiation of human embryonic stem cells (hESCs) into a neural phenotype. The appropriate agent for cell therapy is neural precursors (NPs). Here, we demonstrate the derivation of highly enriched and expandable populations of proliferating NPs from the CCTL14 line of hESCs. These NPs could differentiate in vitro into functionally active neurons, as confirmed by immunohistochemical staining and electrophysiological analysis. Neural cells differentiated in vitro from hESCs exhibit broad cellular heterogeneity with respect to developmental stage and lineage specification. To analyze the population of the derived NPs, we used fluorescence-activated cell sorting (FACS) and characterized the expression of several pluripotent and neural markers, such as Nanog, SSEA-4, SSEA-1, TRA-1 60, CD24, CD133, CD56 (NCAM), beta-III-tubulin, NF70, nestin, CD271 (NGFR), CD29, CD73, and CD105 during long-term propagation. The analyzed cells were used for transplantation into the injured rodent brain; the tumorigenicity of the transplanted cells was apparently eliminated following long-term culture. These results complete the characterization of the CCTL14 line of hESCs and provide a framework for developing cell selection strategies for neural cell-based therapies. PMID- 20021735 TI - Bilirubin promotes de novo generation of T regulatory cells. AB - We have previously demonstrated that bilirubin administration to the recipient induces tolerance towards islet cell transplants across a complete MHC mismatch in a mouse model. Here we assess the mechanisms of such protection. Bilirubin treatment of recipients improved function of islet allografts by suppressing expressions of proinflammatory and proapoptotic genes in those islets and by increasing Foxp3(+) T regulatory (Treg) cells at the site of transplanted islets at various days after transplantation. No prolongation of graft survival was observed in recipients treated with bilirubin when CD4(+)CD25(+) T cells were predepleted from those recipients, indicating that Treg cells are necessary for the protective effect of bilirubin. Adoptive transfer of Treg cells from tolerant mice into Rag1(-/-) recipients resulted in long-term acceptance of skin allografts in an alloantigen-specific manner, suggesting that Treg cells are sufficient to induce tolerance. In addition, bilirubin treatment promoted de novo generation of Treg cells in Rag1(-/-) recipients. Thus, bilirubin treatment to the recipients prolongs islet allograft survival via a Treg-dependent manner in which CD4(+)CD25(+) Treg cells are both necessary and sufficient for tolerance induction and graft acceptance. Bilirubin treatment promotes de novo generation of Treg cells that might account for the protective effects of bilirubin given to recipients. PMID- 20021736 TI - Monocyte derivatives promote angiogenesis and myocyte survival in a model of myocardial infarction. AB - In this study, we have investigated the hypothesis that previously reported beneficial effect of peripheral blood mononuclear cells cultured under angiogenic conditions on cardiovascular function following ischemia is not limited to EPCs but also to monocytes contained therein. We first purified and analyzed the phenotype and secretome of human and murine blood monocytes cultured under angiogenic conditions (named MDs for monocyte derivatives) and tested their effect in a mouse model of myocardial infarction (MI). FACS analysis of MDs shows that these cells express mature endothelial cell markers and that their proliferative capacity is virtually absent, consistent with their end differentiated monocytic ontogeny. MDs secreted significant levels of HGF, IGF-1, MCP-1, and sTNFR-1 relative to their monocyte precursors. MDs were unable to form vascular networks in vitro when cultured on matrix coated flasks. Treatment of murine HL-1 cardiomyocyte cell line with MD-conditioned medium reduced their death induced by TNF-alpha, staurosporine, and oxidative stress, and this effect was dependent upon MD-derived sTNFR-1, HGF, and IGF-1. We further demonstrate that MD secretome promoted endothelial cell proliferation and capacity to form vessels in vitro and this was dependent upon MD-derived MCP-1, HGF, and IGF-1. Echocardiography analysis showed that MD myocardial implantation improved left ventricle fractional shortening of mouse hearts following MI and was associated with reduced myocardial fibrosis and enhancement of angiogenesis. Transplanted MDs and their secretome participate in preserving functional myocardium after ischemic insult and attenuate pathological remodeling. PMID- 20021737 TI - Standardizing optic nerve crushes with an aneurysm clip. AB - OBJECTIVES: Despite the widespread use of optic nerve injury models to simulate central nervous system injury, model protocols vary from laboratory to laboratory, making it difficult to directly compare findings between studies. METHODS: To standardize the optic nerve crush injury model, the commercially available Yasargil aneurysm clip, which provides a consistent clamping force, was used to produce a crush injury to the rat optic nerve. Histology was verified with hematoxylin-eosin. The number of retinal ganglion cells (RGCs) was counted by fluorescent gold dye labeling. RESULTS: Following nerve crush injury, the density of RGCs was substantially reduced in the aneurysm clip-operated group relative to the normal and sham-operated groups, and no discernable difference was noted between the latter two control groups. DISCUSSION: The present findings suggest that Yasargil aneurysm clip effectively produces permanent injury to the optic nerve with evidence from retrograde tracing of RGCs and may provide a standard technique for optic nerve crush studies. PMID- 20021738 TI - Effects of methyl prednisolone acetate, fibrin glue and combination of methyl prednisolone acetate and fibrin glue in prevention of epidural fibrosis in a rat model. AB - OBJECTIVES: Epidural fibrosis, which develops during the post-operative period in 6-20% of the patients who undergo lumbar spinal surgery, can cause persistent low back pain and signs of root compression. Conservative treatment protocols or repeat operations performed for the symptoms of epidural fibrosis are long-term and costly treatments and impairs the patient's quality of life. In this experimental study, we applied methyl prednisolone acetate mixed with fibrin glue to the surgical field and examined the effects on epidural fibrosis in the surgical field by delaying the absorption of methyl prednisolone acetate. METHODS: One hundred Sprague-Dawley rats were divided into five groups, and animals underwent total laminectomy of L4 and L5. We applied 0.05 ml/kg fibrin glue, 0.05 ml/kg methyl prednisolone acetate, 0.05 ml/kg fibrin glue + methyl prednisolone acetate and 0.10 ml/kg fibrin glue + methyl prednisolone acetate topically to the operative sites. Normal saline was applied in the control group. Following the surgery, animals were killed at weeks 1, 2, 4 and 6, and laminectomy sites were examined histopathologically for fibrosis, acute inflammation, necrosis and abscess formation. RESULTS: None of the options had a statistically significant transcendence over others in terms of preventing epidural fibrosis. CONCLUSION: Many biological and non-biological materials have been tried in a quest to prevent epidural fibrosis. However, inducing least amount of injury to the anatomy of the tissues and a very good hemostasis seem to be the most effective methods in the prevention of epidural fibrosis. PMID- 20021739 TI - Long-term safety of combined intracerebral delivery of free gadolinium and targeted chemotherapeutic agent PRX321. AB - OBJECTIVES: While convection enhanced delivery (CED) is an effective delivery method that bypasses the blood-brain barrier, its utility is limited by infusate leakage due to catheter misplacement. Therefore, it is critical to evaluate drug distribution during CED infusion. Gadolinium conjugated to diethylenetriamine penta-acetic acid (Gd-DTPA) is a common, readily available MRI contrast agent, which may be able to predict and actively monitor drug distribution. In this study, we assess the long-term safety and toxicity of intracerebrally infused Gd DTPA along with an experimental targeted agent PRX321. METHODS: Fifty-four immunocompetent rats were implanted with intracerebral cannulas linked to subcutaneously placed osmotic pumps. After pump implantation, the rats were randomized into six groups of nine rats each in order to assess the toxicities of six different concentrations of human serum albumin (HSA) with and without Gd DTPA and PRX321. The rats were monitored clinically for 6 weeks before they were autopsied and assessed for histological toxicity to their central nervous system (CNS). RESULTS: There was one unexplained death in a group infusing low concentration HSA, Gd-DTPA and PRX321. Upon microscopic examination of the CNS in that animal, no unexpected histological toxicity was found. Additionally, there were no signs of clinical or histological toxicity in any of the remaining rats, which all survived until the end of the 6 week observation period. DISCUSSION: Free Gd-DTPA can be safely infused via CED in a pre-clinical animal model. Future studies should include its use in predicting and actively monitoring CED drug infusions in early phase human clinical trials. PMID- 20021741 TI - Acute post-stroke adiponectin in relation to stroke severity, progression and 6 month functional outcome. AB - BACKGROUND: Circulating adiponectin (ADPN) has been inversely associated with the risk of coronary artery disease and ischemic stroke (IS), due to its anti inflammatory and anti-atherosclerotic properties. Recent experimental studies have suggested that ADPN may as well exert cerebroprotective properties in brain ischemia, therefore modifying disease outcome. We investigated whether acute post stroke ADPN in humans might be associated with disease severity, progression and outcome. METHODS: Serum ADPN was measured in 82 consecutive acute IS patients. Severity at presentation and stroke progression within the first week were evaluated according to internationally agreed definitions. Disability and functional outcomes were assessed on months 1, 3 and 6 using the modified Rankin scale (mRS) and Barthel index (BI). Additional data included information on infarct size, mortality, recurrent IS and mental state. RESULTS: Higher ADPN was indicative of greater disability (mRS) on month 1 (OR=1.141, 95% CI=1.012-1.286, p=0.031), but this result was not replicated using the BI. ADPN was not found to be associated with either stroke severity, clinical progression, infarct size, recurrent IS, mortality, mental state, disability or functional outcome during the 6 month follow-up. CONCLUSIONS: Despite previous experimental evidence, serum ADPN measured shortly after an acute IS in humans does not seem to reliably predict disease severity, progression or outcome. The concept that circulating ADPN may beneficially modify long-term outcome of an acute IS may not be the case for human stroke. PMID- 20021740 TI - Study of the prevalence of Parkinson's disease using dopamine transporter imaging. AB - INTRODUCTION: To investigate the prevalence of Parkinson's disease (PD), a three phase study was conducted. METHODS: In phase 1, standardized interviews were performed in a random sample of elderly aged 65 years or older using a questionnaire. In phase 2, neurological examinations were performed to clinically diagnose PD. In phase 3, dopamine transporter (DAT) imaging was performed to support the clinical diagnosis. After the three-phase study, longitudinal clinical observation was performed. RESULTS: A total of 714 subjects participated in the phase-1. Two hundred and twenty-two subjects, scored more than two points, were referred to the movement disorder specialist. Eighteen of these subjects showed overt or equivocal parkinsonian features. Three subjects were clinically diagnosed with possible PD: five with essential tremor with equivocal extrapyramidal signs, eight with frontal-subcortical gait disorder and two with drug-induced parkinsonism. The three subjects with possible PD showed a typical PD pattern of reduced DAT density. DAT density was normal in the other 15 subjects. Results of long-term follow-up supported the diagnoses. The crude prevalence of PD was 0.42 per 100 persons. CONCLUSION: During the clinical evaluation, we encountered a very large proportion of subjects with equivocal parkinsonian features, who posed a diagnostic challenge and a substantial risk of misestimating the prevalence of PD. The combination of DAT imaging and longitudinal clinical observation of equivocal cases enabled us to differentiate PD from other conditions. We suspect that the variation in estimates of the prevalence of PD may be attributable to a considerable proportion of subjects with equivocal parkinsonian features and how they are evaluated. PMID- 20021742 TI - Video-based assessment of operative competency in endoscopic sinus surgery. AB - BACKGROUND: The Accreditation Council for Graduate Medical Education requires that residency programs have an effective plan for assessing their residents' competence. Current evaluation tools used for the assessment of operative competency are subjective, recall based, and do not allow providing formative feedback to the resident. The purpose of this study was to evaluate the feasibility and validity of video-based assessment of endoscopic sinus surgery (ESS) skills. METHODS: Five junior (postgraduate year levels 1-3[PGY-1 to -3]) and three senior (PGY-4 and -5) residents were videotaped as they performed ESS on cadavers in the laboratory. Five faculty members watched the recorded videotapes and rated performance using global and checklist assessment tools for ESS. Evaluators were blinded to the identity of the performers. Rating was made feasible by allowing the raters to fast-forward through the tapes when appropriate. Construct validity was calculated by comparing total scores and score on each item of the instrument across the training levels. For all statistical purposes p < 0.05 was considered significant. RESULTS: Results showed construct validity with senior residents performing better than junior residents. It took an average of 20 minutes (range, 7- 39 minutes) to watch and evaluate a video. Interrater reliability, as measured by interclass correlation coefficient across evaluators, was 0.62. CONCLUSION: The use of videotapes combined with the objective assessment tool is a feasible and valid method for evaluating surgical skills. It has implications for residency programs, because it will increase faculty buy-in and reduce the potential for biased assessment. PMID- 20021745 TI - Cry toxins and proteinase inhibitors in transgenic plants do have non-zero effects on natural enemies in the laboratory: rebuttal to Shelton et al. 2009. PMID- 20021746 TI - Appropriate analytical methods are necessary to assess nontarget effects of insecticidal proteins in GM crops through meta-analysis (response to Andow et al. 2009). PMID- 20021743 TI - Neuropathology of the olfactory mucosa in chronic rhinosinusitis. AB - BACKGROUND: Chronic rhinosinusitis (CRS) is a complex heterogeneous inflammatory disease that affects the nasal cavity, but the pathological examination of the olfactory mucosa (OM) in this disease has been limited. METHODS: Nasal biopsy specimens were obtained from 20 control subjects and 50 CRS patients in conjunction with clinical assessments. Histopathology of these nasal biopsy specimens was performed and immunohistochemistry was used to characterize nonneuronal, neuronal, and inflammatory cells in the OM. These OM characteristics were then evaluated to determine the degree to which pathological features may be related to smell loss in CRS. RESULTS: Histopathological examination of control and CRS OM revealed changes in the normal pseudostratified olfactory epithelium (OE): intermixing of goblet cells, metaplasia to squamous-like cells, and erosion of the OE. Lower percentages of normal epithelium and olfactory sensory neurons were found in CRS OE compared with controls. Relative to other CRS patients, those with anosmia had the greatest amount of OE erosion, the highest density of eosinophils infiltrating the OE, and exhibited the most extensive abnormalities on CT and endoscopic examination, including being significantly more likely to exhibit nasal polyposis. CONCLUSION: Our results suggest that OM pathology observed in nasal biopsy specimens can assist in understanding the degree of epithelial change and sensorineural damage in CRS and the potential for olfactory loss. PMID- 20021747 TI - Overwintering survival of Aphthona beetles (Coleoptera: Chrysomelidae): a biological control agent of leafy spurge released in North Dakota. AB - We evaluated the effects of snow cover and debris cover on overwintering success of Aphthona from 2001 to 2004 in southeast North Dakota. Chill degree-days, soil temperature, and duration of soil temperatures were monitored in field plots. Emergence of Aphthona from soil cores collected in October and held in the laboratory under simulated winter conditions did not differ across treatments. Field emergence of Aphthona was significantly reduced compared with emergence of Aphthona in the laboratory under simulated winter. Snow cover protected overwintering of Aphthona during 3 yr. Overwintering success was 77-94% lower in no-snow plots than in snow plots except in 2002. In 2001, when the soil temperature was as low as -4.4 degrees C, 90% of the Aphthona beetles emerged from snow-covered plots. The ranges of winter temperature and winter period for overwintering Aphthona were determined as -5.0-4.5 degrees C and 56-132 d, respectively, by using linear regression. A negative linear relationship between soil temperature and winter period may help predict the minimum needed overwintering period at a given winter temperature. Warmer temperatures during the winter of 2002 caused snow to melt in the snow plots, which led to excessive moisture that seemed to reduce overwintering success of Aphthona in those plots. Overall emergence was too low to conduct statistical procedures in 2003 and 2004 because of temperature extremes and warm periods during the winters that may have caused Aphthona to break diapause prematurely. PMID- 20021748 TI - Texas is the overwintering source of fall armyworm in central Pennsylvania: implications for migration into the northeastern United States. AB - Fall armyworm, Spodoptera frugiperda (J. E. Smith) (Lepidoptera: Noctuidae), infestations in most of North America arise from annual migrations of populations that overwinter in southern Texas and Florida. Cytochrome oxidase I haplotype profiles within the fall armyworm corn strain, the subgroup that preferentially infests corn (Zea mays L.), can differentiate the Texas and Florida populations. We use this molecular metric to show that fall armyworms in central Pennsylvania originate from Texas, indicating the existence of a migratory pathway from Texas to the northeastern United States. These results were compared with historical trapping data for fall armyworm and another migratory noctuid, corn earworm Helicoverpa zea (Boddie) (Lepidoptera: Noctuidae), in the Pennsylvania and Maryland corn-producing areas to better define lepidopteran migratory pathways. PMID- 20021749 TI - Gypsy moth (Lepidoptera: Lymantriidae) flight behavior and phenology based on field-deployed automated pheromone-baited traps. AB - Populations of the gypsy moth, Lymantria dispar (L.), are extensively monitored in the United States through the use of pheromone-baited traps. We report on use of automated pheromone-baited traps that use a recording sensor and data logger to record the unique date-time stamp of males as they enter the trap. We deployed a total of 352 automated traps under field conditions across several U.S. states over a 5-yr period. In many cases, there was general congruence between male moth capture and the number of recorded events. Although it was difficult to decipher an individual recording event because of the tendency for over-recording, the overall distribution of recorded events was useful in assessing male gypsy moth flight behavior and phenology. The time stamp for recorded events corroborated a previous report of crepuscular gypsy moth male flight behavior, because, although most moths were trapped between 12 and 16 h, there was a consistent period of flight activity between 20 and 22 h. The median male flight duration was 24 d (228 DD, base threshold = 10 degrees C), but there were several traps that recorded flight periods >42 d that could not be explained by overcounting given the congruence between moth capture and the number of recorded events. Unusually long flight periods could indicate the introduction of male moths or other life stages that developed under different climatic conditions. PMID- 20021750 TI - Multicolored Asian lady beetle (Coleoptera: Coccinellidae) activity and wine grape phenology: implications for pest management. AB - We determined the phenology of the multicolored Asian lady beetle, Harmonia axyridis (Pallas), adults in relation to the phenology of wine grapes (Frontenac and Marechal Foch) in Minnesota and Wisconsin vineyards to establish a management window for H. axyridis infestations in wine grapes. In addition, we also assessed the flight activity of H. axyridis in an agricultural landscape. The phenology of berry development and ripening was determined by recording berry size and sugar content of randomly selected berries. The phenology of H. axyridis was determined by tracking its flight activity with yellow sticky cards in vineyards and with a blacklight trap in an agricultural landscape. Berry development and ripening showed three distinct growth periods or phases. The end of growth period I averaged 9 July (Frontenac) and 11 July (Marechal Foch). Veraison, which marks the end of growth period II, averaged 25 July (Frontenac) and 3 August (Marechal Foch). Harvest, the third growth period averaged 18 September for Frontenac and 17 September for Marechal Foch. A major peak of H. axyridis captures occurred between veraison and harvest (i.e., the grape susceptible stage). A similar peak in the summer was observed in the agricultural landscape approximately 10 d before the major peak in the vineyards. PMID- 20021751 TI - Investigating the effect of invasion characteristics on onion thrips (Thysanoptera: Thripidae) populations in onions with a temperature-driven process model. AB - A temperature-driven process model was developed to describe the seasonal patterns of populations of onion thrips, Thrips tabaci Lindeman, in onions. The model used daily cohorts (individuals of the same developmental stage and daily age) as the population unit. Stage transitions were modeled as a logistic function of accumulated degree-days to account for variability in development rate among individuals. Daily survival was modeled as a logistic function of daily mean temperature. Parameters for development, survival, and fecundity were estimated from published data. A single invasion event was used to initiate the population process, starting at 1-100 d after onion emergence (DAE) for 10-100 d at the daily rate of 0.001-0.9 adults/plant/d. The model was validated against five observed seasonal patterns of onion thrips populations from two unsprayed sites in the Riverina, New South Wales, Australia, during 2003-2006. Performance of the model was measured by a fit index based on the proportion of variations in observed data explained by the model (R (2)) and the differences in total thrips days between observed and predicted populations. Satisfactory matching between simulated and observed seasonal patterns was obtained within the ranges of invasion parameters tested. Model best-fit was obtained at invasion starting dates of 6-98 DAE with a daily invasion rate of 0.002-0.2 adults/plant/d and an invasion duration of 30-100 d. Under the best-fit invasion scenarios, the model closely reproduced the observed seasonal patterns, explaining 73-95% of variability in adult and larval densities during population increase periods. The results showed that small invasions of adult thrips followed by a gradual population build-up of thrips within onion crops were sufficient to bring about the observed seasonal patterns of onion thrips populations in onion. Implications of the model on timing of chemical controls are discussed. PMID- 20021752 TI - Colonization of ephemeral water bodies in the Wheatbelt of Western Australia by assemblages of mosquitoes (Diptera: Culicidae): role of environmental factors, habitat, and disturbance. AB - Environmental disturbance may have direct and indirect impacts on organisms. We studied the colonization of ephemeral water bodies by mosquitoes (Diptera: Culicidae) in the Wheatbelt region of southwest Western Australia, an area substantially affected by an expanding anthropogenic salinization. Mosquitoes frequently colonized ephemeral water bodies, responded positively to rainfall, and populated smaller water bodies more densely than larger water bodies. We found that the habitat characteristics of ephemeral water bodies changed in association with salinity. Consequently relationships between salinity and abundance of colonizing mosquitoes were direct (salinity-mosquito) and indirect (salinity-water body characteristics-mosquito). Overall, the structure of mosquito assemblages changed with increasing salinity, favoring an increased regional distribution and abundance of Aedes camptorhynchus Thomson (Diptera: Culicidae), a vector of Ross river virus (RRV; Togoviridae: Alphavirus). We conclude secondary salinization in the Western Australia Wheatbelt results in enhanced vectorial potential for RRV transmission. PMID- 20021753 TI - Effects of grassland succession on communities of orb-weaving spiders. AB - Native grasslands are among the most imperiled of the North American ecosystems, but abandoned agricultural areas may provide suitable habitat for animal taxa that are endemic to grasslands. We studied how species diversity of orb-weaving spiders was influenced by secondary succession of a grassland plant community by monitoring the abundance and species diversity in study plots that were cultivated at 6-yr intervals and left uncultivated in the interim. We tested the hypothesis that local abundance and species diversity of spiders would be positively associated with time since cultivation because plant communities in older habitats would be more architecturally complex. Local abundance of spiders in general was not associated with time since cultivation, but abundance of Mangora gibberosa (Hentz) was positively associated with the abundance of perennial plants. Species richness and diversity of spiders also were positively associated with the abundance of perennial plants and reached a threshold a few years after cultivation. Species diversity of orb-weaving spiders seems to be strongly influenced by species composition of the plant community. Therefore, effective restoration of the structure and function of endemic communities of orb weaving spiders may depend on preserving endemic grassland plant communities. PMID- 20021754 TI - Distribution of tabanids (Diptera: Tabanidae) along a two-sided altitudinal transect. AB - The pattern of horse fly (Diptera: Tabanidae) distribution and correlations among biodiversity, abundance, abiotic factors, and altitude were determined along a two-sided altitudinal transect. The sampling was carried out on five 3-d periods during tabanid seasonal activity. Linen canopy traps with 1-octen-3-ol as an attractant were used at 20 sampling sites along the transect. The results showed that the qualitative composition of tabanid species can be distinguished by altitude and, especially, between southeastern and northwestern mountain slopes. The peaks of horse fly species richness and abundance were indicated at middle elevations of both slopes, where horse fly distributional groups were overlapping and most rare and infrequent species were sampled. All expected species were sampled according to species accumulation curve. The canonical correlation analysis separated species and sampling sites into three clusters; two were positively correlated with the temperature and the wind but differed in sensitivity toward them, and the third cluster was correlated with the humidity. The horse fly distribution was nonhomogenous, and the distributional patterns were only partially determined by altitude and vegetation. The determining environmental variables were different for each slope: temperature and wind for the southern slope (Mediterranean climatic zone) and humidity for the northern slope (continental climatic zone). PMID- 20021755 TI - Sex-biased phoretic mite load on two seaweed flies: Coelopa frigida and Coelopa pilipes. AB - Two hypotheses explain male-biased parasitism. Physiological costs of male sexually selected characteristics can reduce immunocompetence. Alternatively, ecological differences could generate male-biased parasitism. One method of comparing the importance of the two theories is to investigate patterns of phoresy, which are only likely to be generated by ecological rather than immunological differences between the sexes. Here we studied the pattern of phoresy of the mite, Thinoseius fucicola, on two species of seaweed fly hosts, Coelopa frigida and Coelopa pilipes. We found a highly male-biased pattern of phoresy of T. fucicola on both species. These are the first reported instances of sex-biased phoresy in a solely phoretic parasite. We also show the first two cases of size-biased phoresy. We suggest that ecological factors, particularly, male mate searching, generated male biased patterns of phoresy. We highlight the potential importance of studies of phoresy in determining the relative roles of the immunocompetence and ecological theories in generating male-biased parasitism. We suggest that more studies of patterns of phoresy are carried out to allow detailed comparisons with patterns of parasitism. PMID- 20021756 TI - Fluctuations in richness and abundance of social wasps during the dry and wet seasons in three phyto-physiognomies at the tropical dry forest of Brazil. AB - The social wasp nests were quantified in three different plant physiognomies (forested Caatinga, shrubby Caatinga, and agricultural systems) to analyze the effect of environmental seasonality and plant physiognomy on the richness, nest abundance, and species composition of social wasps in the region of tropical dry forest of Brazil. The forested Caatinga physiognomy had the greatest richness of species (S = 16), followed by shrubby Caatinga (S = 13) and by agricultural system (S = 12). The first axis of detrended correspondence analysis (DCA) explained 67.8% of the variability and shows a gradient of the fauna from agricultural system and shrubby Caatinga to forested Caatinga. In the first axis, wet season scores were much higher than those for the dry season in forested Caatinga. The second axis explained 18.7% of the variability and shows a separation of samples collected during the wet or the dry periods in shrubby Caatinga. This separation was less evident in the agricultural system. Variations in nest abundance were more intense in arbustive caatinga (45% decrease in number of active nests in the dry period), moderate in forested Caatinga (24% decrease in number of active nests in the dry period), and low in agricultural systems (8% decrease in the dry period). PMID- 20021757 TI - Effect of an invasive and native ant on a field population of the black citrus aphid (Hemiptera: Aphididae). AB - Invasive ants often enter into facultative mutualisms that frequently lead to outbreaks of the hemipteran partner. Native ants may also enter into similar mutualisms but often these do not lead to outbreaks. However, field studies comparing the impact of an invasive and native ant on a honeydew-producing hemipteran are lacking. We monitored numerical changes of the black citrus aphid, Toxoptera aurantii, tended by adjacent colonies of the invasive Argentine ant, Linepithema humile, and the endemic odorous house ant, Tapinoma sessile, during 2005, 2006, and 2007. Ant-tended aphid numbers were higher than those of untended aphids, with L. humile-tended and T. sessile-tended T. aurantii populations being comparable in 2005 and 2007. However, in 2006, a severe storm, with heavy rainfall, reduced T. sessile and aphid populations in areas occupied by T. sessile, whereas L. humile and aphids tended by L. humile were not reduced. This suggested that T. sessile foraging activity and hemipteran-tending was negatively impacted by severe weather. A laboratory experiment simulating rainfall striking the surface of a leaf showed that T. sessile foraging activity declined sharply under severe simulated rainfall conditions, whereas foraging activity of L. humile did not. Maintaining populations of honeydew-producing Hemiptera across broad climatic conditions may be one mechanism by which L. humile gains a competitive advantage over native ants occupying overlapping niches. PMID- 20021758 TI - Transinfection of Wolbachia in planthoppers: nymphal injection of cultured Wolbachia and infection dynamics. AB - Wolbachia species are intracellular symbionts that cause reproductive alterations in arthropods. Transinfection experiments have been performed in many arthropod species to elucidate the interaction between Wolbachia and a new host. To ease transinfection of this bacterium to new arthropod hosts, we introduced two techniques: nymphal injection instead of embryonic injection and the use of a cultured source of Wolbachia instead of direct transfer from donors to recipients. Wolbachia in the small brown planthopper Laodelphax striatellus was cultivated in a cell line and injected into the nymphal body cavity of the brown planthopper Nilaparvata lugens together with the cells. By using these techniques, two transinfected planthopper lines were obtained. In one line, Wolbachia disappeared after several generations; in the other line, Wolbachia was retained for >7 yr. Infection rates in this latter transinfected line were approximately 80% in early generations after transinjection but decreased to <10% through 40-60 generations. Subsequent selection for Wolbachia-infected females in this line did not increase the infection rate as a temporary effect. Thus, this transinfected line of N. lugens showed cytoplasmic incompatibility, although the incompatibility level was lower than in L. striatellus, the original host. The method of transinfection presented herein is useful for transmitting intracellular symbionts between small arthropod hosts. PMID- 20021759 TI - Functional response of Gonatocerus ashmeadi and the "new association" parasitoid G. tuberculifemur attacking eggs of Homalodisca vitripennis. AB - The functional response of two egg parasitoids, Gonatocerus tuberculifemur (Ogloblin) and G. ashmeadi Girault (Hymenoptera: Mymaridae), to varying densities of Homalodisca vitripennis (Germar) (Hemiptera: Cicadellidae) eggs was studied in the laboratory. G. tuberculifemur showed a type II functional response where parasitism increased at a decreasing rate as host egg density increased from 5 to 40, reaching an asymptote of an upper limit of 9.3 host eggs within the 1-h exposure period. The instantaneous attack rate and handling time as estimated by the random predator equation were 2.28/h and 0.10 h, respectively. In contrast, G. ashmeadi showed a type I functional response where the number of hosts attacked followed a constant linear increase regardless of host density to an upper limit of 11.5 host eggs within the 1-h exposure period. Female G. ashmeadi parasitized on average 4.3 and 3.5 more eggs per hour when presented with 10 and 20 H. vitripennis eggs, respectively, compared with G. tuberculifemur. When 5 or 40 H. vitripennis eggs were offered, there was no significant difference in parasitism rates between parasitoid species. Percentage of female offspring was significantly higher (5%) for G. ashmeadi compared with G. tuberculifemur. Under the experimental conditions used in these evaluations, results suggest that G. tuberculifemur is an inferior parasitoid to G. ashmeadi. PMID- 20021760 TI - Fundamental host range of Pseudophilothrips ichini s.l. (Thysanoptera: Phlaeothripidae): a candidate biological control agent of Schinus terebinthifolius (Sapindales: Anacardiaceae) in the United States. AB - Schinus terebinthifolius Raddi (Sapindales: Anacardiaceae) is a non-native perennial woody plant that is one of the most invasive weeds in Florida, Hawaii, and more recently California and Texas. This plant was introduced into Florida from South America as a landscape ornamental in the late 19th century, eventually escaped cultivation, and now dominates entire ecosystems in south-central Florida. Recent DNA studies have confirmed two separate introductions of S. terebinthifolius in Florida, and there is evidence of hybridization. A thrips, Pseudophilothrips ichini s.l. (Hood) (Thysanoptera: Phlaeothripidae), is commonly found attacking shoots and flowers of S. terebinthifolius in Brazil. Immatures and occasionally adults form large aggregations on young terminal growth (stems and leaves) of the plant. Feeding damage by P. ichini s.l. frequently kills new shoots, which reduces vigor and restricts growth of S. terebinthifolius. Greenhouse and laboratory host range tests with 46 plant species in 18 families and 10 orders were conducted in Parana, Brazil, and Florida. Results of no choice, paired-choice, and multiple-choice tests indicated that P. ichini s.l. is capable of reproducing only on S. terebinthifolius and possibly Schinus molle L., an ornamental introduced into California from Peru that has escaped cultivation and is considered invasive. Our results showed that P. ichini s.l. posed minimal risk to mature S. molle plants or the Florida native Metopium toxiferum L. Krug and Urb. In May 2007, the federal interagency Technical Advisory Group for Biological Control Agents of Weeds (TAG) concluded P. ichini s.l. was sufficiently host specific to recommend its release from quarantine. PMID- 20021762 TI - Life history and life tables of Bactericera cockerelli (Homoptera: Psyllidae) on eggplant and bell pepper. AB - The development, survivorship, and fecundity of the potato psyllid, Bactericera cockerelli (Sulc), fed on eggplant (Solanum melongena L., variety Special Hibush) and bell pepper (Capsicum annuum L., variety Capsitrano) were studied in the laboratory at 26.7 +/- 2 degrees C, 70 +/- 5% RH, and at a photoperiod of 14:10 (L:D) h. Immature B. cockerelli developed faster (24.1 d) when fed on eggplant than on bell pepper (26.2 d). Survival rates of immature stages from egg to adult emergence were higher on eggplant (50.2%) than on bell pepper (34.6%). The longevity of B. cockerelli female adults fed on bell pepper was similar to that of females fed on eggplant (62.2 versus 55.0 d), but the male adults fed on eggplant lived shorter lives (39.4 d) than those fed on bell pepper (53.9 d). However, the preoviposition and oviposition periods, fecundity, and sex ratio of B. cockerelli fed on eggplant were not different from those fed on bell pepper. The r(m ) value and the finite rate of increase (lambda) of B. cockerelli were higher on eggplant (0.1099 and 1.116, respectively) than on bell pepper (0.0884 and 1.0924, respectively). Mean generation time and doubling time of B. cockerelli were shorter on eggplant (40.4 and 6.3 d, respectively) than on bell pepper (46.1 and 7.8 d, respectively). In contrast, lifetime fecundity of B. cockerelli was greater on bell pepper (227.3 offspring) than on eggplant (186.5 offspring). Based on these life history parameters, we concluded that B. cockerelli performed better on eggplant than on bell pepper. PMID- 20021761 TI - Differences in preference and performance of the water lily leaf beetle, Galerucella nymphaeae populations on native and introduced aquatic plants. AB - Plant invasions represent ecological opportunities for herbivorous insects able to exploit novel host plants. The availability of new hosts and rapid adaptations may lead to host race formation and ultimately speciation. We studied potential host race formation in the water lily leaf beetle, Galerucella nymphaeae, in response to invasion by water chestnut, Trapa natans, in eastern North America. This leaf beetle is well suited for such studies because previous work showed that different herbivore populations follow different "evolutionary pathways" and specialize locally in response to differences in habitat preferences and host plant availability. We compared host preference and performance of G. nymphaeae offspring originating from T. natans and offspring of individuals originating from an ancestral host Nuphar lutea, yellow water lily, on T. natans and three native hosts (N. lutea, Nympheae odorata, and Brasenia schreberi). Regardless of origin (Trapa or Nuphar), adults strongly preferred their native host, N. lutea, over T. natans. Although laboratory survival rates (larva to pupa) were extremely high (80%) regardless of origin or host offered, survival rates in a common garden were greatly reduced, particularly for T. natans (24%) and to a lesser extent on N. lutea (54%), regardless of beetle origin. Larval drowning during more frequent leaf changes when developing on small Trapa leaves seems to be responsible for this difference. Preference of females for N. lutea is beneficial considering the much higher larval survival on the ancestral host. Abundant T. natans where the plant is invasive provides an alternative food source that beetles can use after egg/larval loads on their preferred host reach carrying capacity, but this utilization comes at a cost of high larval mortality. PMID- 20021763 TI - Emerald ash borer (Coleoptera: Buprestidae) attraction to stressed or baited ash trees. AB - Emerald ash borer, Agrilus planipennis Fairmaire (Coleoptera: Buprestidae), has killed millions of ash (Fraxinus sp.) trees in North America since its discovery in Michigan in 2002. Efficient methods to detect low-density A. planipennis populations remain a critical priority for regulatory and resource management agencies. We compared the density of adult A. planipennis captured on sticky bands and larval density among ash trees that were girdled for 1 or 2 yr, wounded, exposed to the stress-elicitor methyl jasmonate, baited with Manuka oil lures, or left untreated. Studies were conducted at four sites in 2006 and 2007, where A. planipennis densities on untreated trees ranged from very low to moderate. In 2006, 1-yr girdled trees captured significantly more adult A. planipennis and had higher larval densities than untreated control trees or trees treated with methyl jasmonate or Manuka oil. Open-grown trees captured significantly more A. planipennis beetles than partially or fully shaded trees. In 2007, A. planipennis population levels and captures of adult A. planipennis were substantially higher than in 2006. The density of adults captured on sticky bands did not differ significantly among canopy exposure classes or treatments in 2007. Larval density was significantly higher in untreated, wounded, and 1-yr girdled trees (girdled in 2007) than in 2-yr girdled trees (girdled in 2006), where most phloem was consumed by A. planipennis larvae the previous year. A total of 36 trees (32 in 2006, 4 in 2007) caught no beetles, but 16 of those trees (13 in 2006, 3 in 2007) had A. planipennis larvae. In 2006, there was a positive linear relationship between the density of adults captured on sticky bands and larval density in trees. Our results show that freshly girdled and open grown trees were most attractive to A. planipennis, especially at low-density sites. If girdled trees are used for A. planipennis detection or survey, debarking trees to locate larval galleries is crucial. PMID- 20021764 TI - Surplus nectar available for subalpine bumble bee colony growth. AB - Mutualisms may cause coupled population expansion or decline if both partners respond to variation in the other's abundance. Many studies have shown how the abundance of animal mutualists affects plant reproduction, but less is known about how the abundance of plant mutualists affects animal reproduction. Over 2 yr, I compared reproduction of the bumble bee, Bombus appositus, across meadows that varied naturally in flower density, and I compared reproduction between fed colonies and unfed control colonies. Colony reproduction (gyne, worker, and male production) was constant across meadows that varied naturally in flower density. Forager densities per flower did not vary among meadows, and daily nectar depletion was consistently low across meadows, suggesting that bees had ample nectar in all meadows. However, colonies directly fed with supplemental nectar and pollen generally produced over twice as many gynes as control colonies. Feeding did not affect male or worker production. Although colonies may benefit from food supplementation at the nest, it is possible that they may not benefit from additional flowers because they have too few workers to collect extra resources. PMID- 20021765 TI - Determining circadian response of adult male Acrobasis nuxvorella (Lepidoptera: Pyralidae) to synthetic sex attractant pheromone through time-segregated trapping with a new clockwork timing trap. AB - Mate finding is a key lifecycle event for the pecan nut casebearer, Acrobasis nuxvorella Neunzig, as it is for virtually all Lepidoptera, many of which rely on long-range, species-specific sex pheromones, regulated largely by circadian clocks. Adult male moths were trapped at discrete time intervals during the first two seasonal flights for 6 yr to determine times of peak activity associated with male response to pheromones. From 1997 to 2002, the Harris-Coble automated clockwork timing trap was used for hourly time-segregated sampling. Analysis of variance with linear contrasts determined that circadian response of A. nuxvorella males to pecan nut casebearer pheromone began at approximately 2100 hours, the first hour of total darkness, lasting for 6-7 h. It peaked from midnight to 0400 hours and ended at the onset of morning twilight, approximately 0500 hours. The hours of peak activity are hours of minimal bat predation. The study shows that pecan nut casebearer males become responsive to pheromone several hours before females start calling and remain responsive for at least 1 h after they stop. The extended response period conforms to studies of other polygamous Lepidoptera in which a selective advantage is conferred on early responding males in scramble competition for available females. PMID- 20021766 TI - Flight activity of two sympatric termite species, Macrotermes gilvus and Macrotermes carbonarius (Termitidae: Macrotermitinae). AB - Flight activities of two sympatric termite species, Macrotermes gilvus (Hagen) and Macrotermes carbonarius (Hagen), were studied in Penang Island, Malaysia. Herein, we present the first documentation of chronological reproductive isolation of M. gilvus and M. carbonarius. Flights of M. gilvus were recorded over a remarkably long 7-mo period from March to September, whereas swarming of M. carbonarius took place from November to January. Swarming events of M. gilvus and M. carbonarius occurred under atmospheric pressures of 1,005-1,011 and 1,006 1,010 hPa, respectively. Most flights of M. gilvus occurred on days with rain, whereas M. carbonarius avoided rain. Flight activity of M. gilvus was correlated significantly with atmospheric pressure and rainfall. The threshold temperature and relative humidity of M. gilvus flights were between 23 and 26 degrees C and 83 and 98% RH, respectively; M. carbonarius swarmed between 25 and 30 degrees C and 72 and 83% RH, respectively. The flight activity of M. gilvus concentrated in the warmer and humid months with a monthly total rainfall of 228 mm. Both species swarmed at distinct times of day during the limited field observations: Flights of M. gilvus began between 0300 and 0430 hours (light intensity <1 Lx), and flights of M. carbonarius lasted for only 4-10 min between 1900 and 1910 hours (at dusk; light intensity: 20-200 Lx). Windless conditions were preferred for the flights of both species. PMID- 20021767 TI - Oviposition behavior of the wheat stem sawfly when encountering plants infested with cryptic conspecifics. AB - Insect herbivores typically oviposit on the most suitable hosts, but choices can be modulated by detection of potential competition among conspecifics, especially when eggs are deposited cryptically. Larvae of the wheat stem sawfly, Cephus cinctus Norton, developing within an already infested stem, experience elevated risk when only one will survive because of cannibalism. To increase our understanding of host selection when the choices made by females can lead to severe intraspecific competition, females were presented with either uninfested wheat plants or with plants previously exposed to other females in laboratory choice tests. The oviposition behavior of this insect was described by recording the behavioral sequences that lead to and follow the insertion of the ovipositor in both previously infested and uninfested stems. No significant differences were found in frequencies of specific behaviors or behavioral transitions associated with oviposition. In choice tests, there was no difference in the numbers of eggs laid in infested and uninfested plants. Taller plants received more eggs, irrespective of infestation. Females neither preferred nor avoided previously infested hosts. Other characteristics of the host, such as stem height, may be more important in determining suitability for oviposition. These findings support the use of management tactics relying on the manipulation of oviposition behavior, such as trap cropping. Given that there is no evidence for response to previously infested hosts, the infested plants in a trap crop would remain as suitable as they were when uninfested, which could also lead to an increase in mortality caused by intraspecific competition. PMID- 20021768 TI - Predicting black light trap catch and flight activity of Acrosternum hilare (Hemiptera: Pentatomidae) adults. AB - A regression model was developed to predict the flight activity of Acrosternum hilare (Say) using data on the number of adults collected in a single black light trap located in Painter, VA, in the 18-yr period from 1990 to 2007. Eighteen initial weather variables, including cumulative precipitation over different time periods, mean monthly precipitation (PJA) and days below freezing (DFJA) from January to April, and mean monthly temperatures from December to April were tested in developing the regression model. Mixed (backward and forward) stepwise regression analysis showed that a two-variable model using PJA and DFJA was adequate for predicting the seasonal mean weekly number of A. hilare adults in the trap. Validation of the model using five independent black light trap data sets resulted in a strong correlation (r = 0.98) between observed and predicted mean weekly number of A. hilare adults caught in traps. Three peaks in flights of A. hilare adults were observed when mean trap catch was plotted over time for the 18-yr period. Peaks occurred at 319, 892, and 1,331 degree days (DD) from 1 January. Based on known developmental rates, the first peak was attributed to overwintered adults, the second to first-generation adults, and the third to a second generation of adults. This research suggests that A. hilare undergoes two complete generations in Virginia. Cumulative trap catch estimated from the 18-yr mean trap catch showed that 10, 50, and 90% of the total seasonal catch should occur by 153, 501, and 1,066 DD. PMID- 20021769 TI - Effectiveness and biases of Winkler litter extraction and pitfall trapping for collecting ground-dwelling ants in northern temperate forests. AB - The sampling efficiency of pitfall traps and Winkler litter extraction in northern deciduous forests was compared using ants. Both techniques are among the most common methods used to measure the diversity of organisms active on the forest floor. During 2005-2006, 90 Winkler and 180 pitfall trap samples from urban forest fragments in northeastern Ohio obtained 9,203 ants representing 31 species. Winklers captured all 31 species, whereas pitfall traps collected a total of 24 species. Winkler samples accumulated species more rapidly than did pitfall traps and had greater total species richness and higher abundance of ants recorded. Consistent with other studies, Winkler sampling was found to catch a greater number of smaller ants, whereas pitfall trapping caught a greater number of large-bodied ants. According to estimates of expected species richness, the combination of the two sampling techniques allowed for the collection of approximately 90% of the ants expected in the surveyed area. Site variation had little effect on the inherent differences in sampling efficacy between the two methods. Either technique adequately collected samples for broad comparisons and documentation of the more typical and representative ant fauna, but Winkler extraction exhibited the advantage of a more complete inventory. The application of both techniques should be considered if the aims of a study require estimation of community properties, such as relative abundance. PMID- 20021770 TI - Thresholds, injury, and loss relationships for thrips in Phleum pratense (Poales: Poaceae). AB - Timothy (Phleum pratense L.) is an important forage crop in many Western U.S. states. Marketing of timothy hay is primarily based on esthetics, and green color is an important attribute. The objective of these studies was to determine a relationship between arthropod populations, yield, and esthetic injury in timothy. Economic injury levels (EILs) and economic thresholds were calculated based on these relationships. Thrips (Thripidae) numbers were manipulated with insecticides in small plot studies in 2006, 2007, and 2008, although tetranychid mite levels were incidentally flared by cyfluthrin in some experiments. Arthropod population densities were determined weekly, and yield and esthetic injury were measured at each harvest. Effects of arthropods on timothy were assessed using multilinear regression. Producers were also surveyed to relate economic loss from leaf color to the injury ratings for use in establishing EILs. Thrips population levels were significantly related to yield loss in only one of nine experiments. Thrips population levels were significantly related to injury once before the first annual harvest and twice before the second. Thrips were the most important pest in these experiments, and they were more often related to esthetic injury rather than yield loss. EILs and economic thresholds for thrips population levels were established using esthetic injury data. These results document the first example of a significant relationship between arthropod pest population levels and economic yield and quality losses in timothy. PMID- 20021771 TI - Attraction of Anoplophora glabripennis to male-produced pheromone and plant volatiles. AB - The male-produced pheromone of Anoplophora glabripennis (Motschulsky) (Coleoptera: Cerambycidae), which is an equal blend of 4-(n-heptyloxy)butan-1-ol and 4-(n-heptyloxy)butanal, was used in laboratory bioassays and in the greenhouse to determine its potential for attracting A. glabripennis adults. In modified "walking wind tunnels," virgin females were most attracted to the alcohol component, and virgin males were repelled by the pheromone blend at the lowest and highest amounts offered. Y-tube olfactometer bioassays also showed that females were significantly more attracted to the pheromone and its components than males were. However, males were more attracted to plant volatiles than females. Of 12 plant volatiles tested, delta-3-carene and (E)-caryophyllene were highly attractive to males, whereas (Z)-3-hexenyl acetate was repellent to males. Combining the male pheromone blend with (-)-linalool alone or with (Z)-3 hexen-1-ol attracted significantly more males than did the pheromone alone. We tested four trap designs in our quarantine greenhouse with eight different lures. The Intercept Panel traps and the hand-made screen sleeve traps caught more beetles than the Plum Curculio traps and Lindgren funnel traps. Intercept traps worked best when baited with male blend and (Z)-3-hexen-1-ol, whereas screen sleeve traps were most attractive when baited with (-)-linalool. Our findings provide evidence of the attractiveness of the A. glabripennis male-produced pheromone and suggest that it has a role in mate-finding. It is also a first step toward the development of an efficient trap design and lure combination to monitor A. glabripennis infestations in the field. PMID- 20021772 TI - Biotic and abiotic factors affect green ash volatile production and emerald ash borer adult feeding preference. AB - The emerald ash borer, Agrilus planipennis Fairmaire (Coleoptera: Buprestidae), is an exotic woodborer first detected in 2002 in Michigan and Ontario and is threatening the ash resource in North America. We examined the effects of light exposure and girdling on green ash (Fraxinus pennsylvanica Marsh) volatile production, and effects of light exposure, girdling, and leaf age on emerald ash borer adult feeding preferences and phototaxis. Green ash seedlings grown under higher light exposure had lower amounts of three individual volatile compounds, (Z)-3-hexenol, (E)-beta-ocimene, and (Z,E)-alpha-farnesene, as well as the total amount of six detected volatile compounds. Girdling did not affect the levels of these volatiles. Emerald ash borer females preferred mature leaves, leaves from girdled trees, and leaves grown in the sun over young leaves, leaves from nongirdled trees, and leaves grown in the shade, respectively. These emerald ash borer preferences were most likely because of physical, nutritional, or biochemical changes in leaves in response to the different treatments. Emerald ash borer females and males showed positive phototaxis in laboratory arenas, a response consistent with emerald ash borer preference for host trees growing in sunlight. PMID- 20021773 TI - Development, reproduction, and within-plant infestation patterns of Aphis craccivora (Homoptera: Aphididae) on alfalfa. AB - The goal of this research was to describe developmental rates, reproductive rates, and infestation patterns of Aphis craccivora Koch on alfalfa (Medicago sativa L.). All studies were conducted on the susceptible cultivar OK08 using aphids reared from collections made in Oklahoma. To determine thermal requirements for growth of A. craccivora, development from birth to adult was recorded at 7.2, 12.8, 18.3, 23.9, and 29.4 degrees C. The same constant temperature treatments (except for 7.2 degrees C being raised to 8.3 degrees C) were used to assess the influence of temperature on reproductive rates. Within plant distribution patterns were determined by infesting three stems on each of 24 plants and recording numbers of A. craccivora on leaf blades, petioles, and internodal stems sections at 2-d intervals through 10 d after infestation. Aphid counts were analyzed to determine significant differences among node parts (leaf blades, petioles, and stem sections). The developmental threshold temperature for A. craccivora was calculated to be 7.1 degrees C, and the thermal constant for development from the first instar to reproducing adult was 100 DD ( degrees C). The optimal temperature range for reproduction on alfalfa was 18-24 degrees C, with a mean of 82 nymphs produced per female. From the initial infestation of three apterae per stem, numbers increased to a mean of 510 per stem after 10 d. Plant profiles showed that the greatest numbers of aphids were located in middle and lower portions of the plant canopy. On all sampling dates, the proportion of aphids on internodal stem sections was significantly greater (P < 0.05) than on petioles and leaf blades. PMID- 20021774 TI - Complexity loss in physiological time series of patients in a vegetative state. AB - Consciousness has not yet been satisfactorily defined because of its puzzling nature which involve the perception of the environment (perceptual awareness) and of the self (self-awareness). Current available methods fail in establishing prognosis in patients with vegetative state (VS): to our mind, this failure stems from the heterogeneous localization of brain damages causing VS and from available approaches tending to investigate self-awareness separately from perceptual awareness, whereas consciousness should be explored as a single and indivisible whole. Moving from the assumption that consciousness depends on the normal activity of wide neural networks, that may be regarded as complex systems whose outputs show a nonlinear behaviour, we propose a nonlinear approach applied to electroencephalographic (EEG) signal, aimed at exploring residual neural networks complexity in patients with VS. For this objective the EEG recording of 10 patients previously admitted to our department were retrospectively analyzed and compared with those of ten matched healthy control subjects. Approximate Entropy (ApEn) was calculated from the average values of time series with fixed input variables. Mean ApEn values were lower in patients than in controls (t18 18 = 12.3, p < 0.001). ApEn is able to discriminate patients from controls thus supporting the hypothesis about a decreased neural networks complexity in VS. PMID- 20021775 TI - Evidence of reduced complexity in self-report data from patients with medically unexplained symptoms. AB - Physical symptoms which cannot be adequately explained by organic disease are a common problem in all fields of medicine. Reduced complexity, shown using nonlinear dynamic analysis, has been found to be associated with a wide range of illnesses. These methods have been applied to short time series of mood but not to self-rated physical symptoms. We tested the hypothesis that self-reported medically unexplained physical symptoms display reduced complexity by measuring the approximate entropy of self-reported emotions and physical symptoms collected twice daily over 12 weeks and comparing the results with series-specific surrogate data. We found that approximate entropy (ApEn) was lower for actual data series than for surrogate data. There was no significant difference in entropy between different types of symptoms and no significant correlation between entropy and the diurnal variation of the data series. Future studies should concentrate on specific symptoms and conditions, and evaluate the effect of treatment on the entropy of symptom patterns. PMID- 20021777 TI - How nonlinear systems inform meaning and early education. AB - This paper contends that educators need to prepare young learners to function with the predictable unpredictability of life in this century by participating in the kind of education that is transformational and adaptive to the ways in which children acquire meaning. When teachers implement a dynamic-themes theory of early education they can help children younger than nine years of age to integrate meaning. The isomorphic relationships apparent within each of several theories--script theory, Theory of Mind as metacognition, and chaos and complexity theory--together can provide evidence for the nature of meaning and its relationship to the nonlinear early education of children. The transformational-generative characteristics of these theories, that include constructed learning, inform dynamic early educational practice. Play, as one condition for meaningful early learning, also has particular significance in early education by affording an environment for assessing learning. PMID- 20021776 TI - Oscillations in daily pain prediction accuracy. AB - Dynamical systems modeling was used to analyze fluctuations in the pain prediction process of people with rheumatoid arthritis. 170 people diagnosed with rheumatoid arthritis completed 29 consecutive days of diaries. Difference scores between pain predictions and next-day pain experience ratings provided a time series of pain prediction accuracy. Pain prediction accuracy oscillated over time. The oscillation amplitude was larger at the start of the diary than at the end, which indicates damping toward more accurate predictions. State-level psychological characteristics moderated the damping pattern such that the oscillations for patients with lower negative affect and higher pain control damped more quickly than the oscillations for their counterparts. Those findings suggest that low negative affect and high pain control generally contributed to a more accurate pain prediction process in the chronically ill. Positive affect did not differentiate the damping pattern but, within each oscillation cycle, patients with higher positive affect spent more time making inaccurate predictions than their counterparts. The current analyses highlight the need to account for change in data through dynamical modeling, which cannot be fully observed through traditional statistical techniques. PMID- 20021778 TI - Numerical explorations of R. M. Goodwin's business cycle model. AB - Goodwin's model, which was formulated in , still attracts economists' attention. The model possesses numerous interesting properties that have been discovered only recently due to the development of the chaos theory and the complexity theory. The first numerical explorations of the model were conducted in the early s by Strotz, McAnulty and Naines (1953). They discovered the coexistence of attractors that are well-known today, two properties of chaotic systems: the sensitive dependence on the initial conditions and the sensitive dependence on parameters. The occurrence of periodic and chaotic attractors is dependent on the value of parameters in a system. In case of certain parametric values fractal basin boundaries exist which results in enormous system sensitivity to external noise. If periodic attractors are placed in the neighborhood of the fractal basin boundaries, then even a low external noise can move the trajectory into the region in which the basin's structure is tangled. This leads to a kind of movement that resembles a chaotic movement on a strange attractor. In Goodwin's model, apart from typical chaotic behavior, there exists yet another kind of complex movements - transient chaotic behavior that is caused by the occurrence of invariant chaotic sets that are not attracting. Such sets are represented by chaotic saddles. Some of the latest observation methods of trajectories lying on invariant chaotic sets that are not attracting are straddle methods. This article provides examples of the basin boundary straddle trajectory and the saddle straddle trajectory. These cases were studied by Lorenz and Nusse (2002). I supplement the results they acquired with calculations of capacity dimension and correlation dimension. PMID- 20021779 TI - Asset price and wealth dynamics as an adaptive belief system. AB - In this paper we study an asset pricing model with heterogeneous beliefs, wealth dynamics and time varying proportions. We derive the resulting deterministic nonlinear dynamic system and analyze the model with adaptive beliefs and switching mechanism in order to investigate bifurcations and global dynamics and thus consider the effects on wealth distribution among different types of agents. Our study aims at confirming that heterogeneity in beliefs may lead to complexity. PMID- 20021780 TI - Chaos, archetypes, and the all-integrating field. AB - This essay explores the notion that the fundamental ontology of the universe is an all-integrating field. The process of chaos operates within this field as do other phenomena such as gravity and electromagnetic force. I argue that there are 'no objects only events', that is, dynamic continual process is the basic building block of all existence. My accompanying artworks illustrate these principles using the beauty of computer generated fractals combined with digitally manipulated, figurative images. PMID- 20021781 TI - [Consideration and suggestions on screening for neonatal diseases in China]. PMID- 20021782 TI - [To establish the classification and admittance system of neonatal intensive care units in China]. PMID- 20021783 TI - [Hemolytic disease of neonates due to anti-M: report of one case and review of reports of 21 cases]. AB - OBJECTIVE: To analyze and summarize clinical manifestation of hemolytic disease of the newborn (HDN) due to anti-M. METHODS: Data of one case of HDN due to anti M and the reports of 21 cases seen in the past 20 years at the home country were reviewed and analyzed. RESULTS: There was an increasing number of reports of cases with HDN due to anti-M. Among the 22 cases, four were the first fetus. Of 18 infants, ten were male, and eight were female. The blood group was MN in 19/21 infants, and was M in 2/21 infants. The blood group was N in 10/21 mothers, and was NN in 11/21 mothers. Among the 18 infants, the direct antiglobulin test of 7 infants were positive, of 4 infants were dubiously positive, and of 7 infants was negative. Among the 16 infants, the antibody release test of 13 infants was positive, and of 3 infants were negative. Among 17 infants, the free antibody test of all was positive. Among the 21 mothers, the anti-M of IgG were positive in all mothers, and along with IgM in 11 mothers. The anti-M of IgG was positive in all infants. Mild or severe anemia and icterus were found in all cases. Among the 15 cases, jaundice was evident on the 1st day of life in 11 cases. Among 13 cases, marked elevation of both indirect- and direct-reacting bilirubin levels was reported in 4 cases. Phototherapy was applied when jaundice became evident. High-dose intravenous immunoglobulin was given to 4/15 cases. Exchange transfusion were performed in 8 of 22 cases. Three cases died, and 19 cases were cured. CONCLUSION: HDN of varying degrees of severity has been reported in association with anti-M and can even lead to intrauterine deaths or requiring treatment with exchange transfusion. If the mother has a history of prior intrauterine deaths, abortion, hydrops fetalis, severe fetal anemia or infertile, MN blood group and anti-M antibodies should be tested after excluding the possibility of other causes and HDN due to ABO or Rh blood group incompatibility. As the efficacy of phototherapy increases, the role of exchange transfusion in acute management is rapidly decreasing. High-dose intravenous immunoglobulin and/or intramuscular metalloporphyrins may further reduce the need for exchange transfusion. The exchange transfusion may be performed through peripheral arterial (drawn out) and venous (infused in) lines. PMID- 20021785 TI - [Gene mutation analysis of a collodion baby]. AB - OBJECTIVE: To determine the mutations pattern of the genes of a collodion baby. METHODS: Collodion baby is a genetic heterogeneous disease caused by mutations of several genes. Since the most common mutations were observed in TGM1 gene, this gene was chosen for mutation screening. The screening was carried out by PCR and direct sequencing. The allele specific primers were designed for a missense mutation and allele-specific (AS) PCR was carried out in 50 normal individuals for population study. RESULTS: Three novel alterations were detected in TGM1 gene of the proband, a missense mutation c.463C > T (p.Arg155Trp) in exon 3, a nonsense mutation c.578G > A (p.Trp193X) in exon 4, and a single nucleotide deletion (c.694delG) also in exon 4 of TGM1 gene. This infant's father was heterozygote of c.694delG mutation, while his mother carried the two mutations (c.463C > T and c.578G > A) on the same chromosome. The missense mutation was not detected in his father and in any of the control individuals by AS-PCR. CONCLUSION: Three novel mutations were identified in TGM1 gene in a Chinese collodion baby. A double mutation (c.463C > T and c.578G > A) located on the maternal allele while the c.694delG deletion on the paternal allele. PMID- 20021786 TI - [Retrospective analysis of elective caesarean section and respiratory distress syndrome in the term neonates]. AB - OBJECTIVE: Severe respiratory distress syndrome (RDS) caused by pulmonary surfactant (PS) deficiency is described not only in preterm infants but also in term babies delivered via caesarean section, especially before the onset of labour (elective caesarean section). Once RDS of term neonates happened, mechanical ventilation is needed, and the infants were at high risk of developing further complications such as persistent pulmonary hypertension of neonates (PPHN), pulmonary air leak and cardiovascular instability, even fatal outcome cannot be avoided. The present study aimed to analyze the association between the elective caesarean section and respiratory distress syndrome (RDS) in term neonates, and to determine the related factors and outcomes of RDS cases in neonatal intensive care unit (NICU) and neonatology ward. METHODS: A retrospective study was conducted at the NICU and the Neonatology Ward of A center (Children's Hospital of Zhejiang University) and the NICU of center B (Obstetrics and Gynecology Hospital of Zhejiang University) on 90 term infants who were diagnosed as RDS between June 2006 and June 2008. The general clinical data, mode of delivery, severity of the radiological sign, pulmonary surfactant (PS) application, the onset time and duration of mechanical ventilation, the ratio of PaO(2) to FIO(2) before mechanical ventilation, oxygenation index (OI), duration of oxygen supplementation, the length of hospital stay and complications including pulmonary air leaks (pneumothorax, pulmonary interstitial emphysema), PPHN, systemic hypotension and ventilator-associated pneumonia (VAP) were collected. The gestational age distribution was studied in RDS cases delivered by elective caesarean section, and the comparative analysis and non-conditional logistic regression analysis wer performed for clinical characteristics and risk factors between the RDS cases with or without complications. P < 0.05 was considered to be significant. RESULTS: There were 88 episodes of elective caesarean section in 90 RDS patients. The proportion of elective caesarean section was 97.8% (88/90). The number of RDS cases was reduced gradually with the increase of gestational age and the constituent ratio of RDS at 39 w and at 40 w was significantly lower than that of 37 w and 38 w. By logistic regression analysis, the onset time of mechanical ventilation > 12 h independently predicted complications in RDS of term neonates (odds ratio 12.667, 95% confidence interval, i.e., CI 1.455 to 110.300, P = 0.021). Moreover, there was a significant difference in the admission age (t = 11.833, P = 0.001), severity of the radiological findings (t = 4.85, P = 0.028), PS application (t = 11.911, P = 0.002), the onset time of mechanical ventilation (t =10.051, P = 0.018), the ratio of PaO(2) to FIO(2) before mechanical ventilation (chi(2) = 4.184, P = 0.005), OI > 25 (t = 4.737, P = 0.03), duration of oxygen supplementation (chi(2) = 10.475, P = 0.001), systemic hypotension (t = 11.020, P = 0.01) and the length of hospital stay (t = 9.827, P = 0.002) between the two centers. CONCLUSION: Severe RDS can occur in term babies after elective caesarean section, gestational age at the time of elective caesarean section may also be important for RDS in term neonates. The onset time of mechanical ventilation > 12 h independently predicted complications in RDS of term neonates. The main complications of RDS in term neonates were PPHN, pulmonary air leaks and systemic hypotension. Early diagnosis, early intervention can significantly reduce the complications, alleviate the severity and shorten their time for oxygen therapy and their length of stay in NICU in term RDS infants. PMID- 20021787 TI - [Evaluation on the early hemodynamic changes after cardiac surgery for congenital heart diseases in neonates]. AB - OBJECTIVE: To accurately evaluate the early hemodynamic status of neonates who undergo complex neonatal cardiac surgery, through monitoring the cardiac index (CI), serum lactate (Lac), mixed venous oxygen saturation (SvO(2)). METHODS: From January to November 2007, haemodynamic data of 80 patients who had open heart surgery for congenital heart disease were analyzed within 48 hours after operation. Of the 80 patients, 47 were neonates, their age ranged from 3 days to 29 days [mean (21.98 + or - 8.15) days] and weight ranged from 2.6 kg to 4.2 kg [mean (3.51 + or - 0.39) kg]. As the control group, 33 young infants at the age of 30 days to 180 days [mean (76.36 + or - 24.79) days] with body weight ranged from 3.1 kg to 6.0 kg [mean (4.59 + or - 0.59) kg] were also enrolled. The value of CI derived from pulse contour and was calculated by using the PiCCO system. Meanwhile, measurements of serum lactate level and SvO(2) were recorded. Serial measurements of the cardiac output were performed for the neonates. RESULTS: CI in survivors of neonates (2.01 + or - 0.35) L/(min x m(2)) was lower than that of the infants (2.26 + or - 0.39) L/(min x m(2)) after cardiac surgery (P < 0.05) at 2 h, 6 h postoperatively. However, urine output remained normal. The value of pulse pressure in neonates was less than that in young infants. Serum lactate level in neonates was significantly higher than that of young infants during cardiac surgical procedures (P < 0.01) at 12 h postoperatively; the SvO(2) was more than 60% postoperatively in survived neonates, there was no significant difference (P > 0.05) in SvO(2) between neonates and young infants during preoperative and postoperative periods. There was a positive correlation between CI and SvO(2). Four neonates and 1 young infant died after surgical treatment, surgical mortality was 8.5% and 3.0%, respectively. The deaths of the neonates were related to the cardiocirculatory function decompensation, unrelieved severe acidosis preoperatively, and the transposition of great artery with coronary artery malformation and longer cardiopulmonary bypass. The patients with significantly high arterial blood lactate levels during the first 6 - 12 hours postoperatively had poor outcome. Lactate levels were higher than 10 mmol/L and SvO(2) less than 50% in neonates who developed multiple organ system failure. One young infant died of sudden arrhythmia after surgical treatment, whose death may be related the surgical procedure itself with pulmonary artery banding and blalock-taussig shunt leading to increased preload and afterload of the heart. CONCLUSIONS: Elevated serum lactate level postoperatively may reflect intraoperative tissue hypoperfusion. Serial blood lactate level measurements may be an accurate predictor of clinical outcomes in children after pediatric open heart surgery. Mixed venous oxygen saturation changes more rapidly than other standard hemodynamic variables. The higher mortality of neonates with congenital heart disease is related to the malformation complexity itself and illness severity. PMID- 20021788 TI - [Effect of intrauterine lipopolysaccharide infusion on Toll-like receptor 4 signaling transduction pathway in lungs of perinatal rats]. AB - OBJECTIVE: To investigate the influence of intrauterine infection caused by lipopolysaccharide on Toll-like receptor 4 (TLR4) signaling pathway in fetal and neonatal rat lungs in order to explore immunomodulating activity of innate immunity responding to intrauterine infection and its effect on lung development. METHODS: On day 17 of pregnancy, 30 pregnant Sprague-Dawley (SD) rats were randomly divided into two groups: LPS group and saline group. For LPS group, LPS (10 microl, 40 microg/ml) was intrauterine injected between every two embryonic sacs of the pregnant rats, while the rats in the control group were injected with the same volume of pyrogen-free saline. Lung tissues of fetal rats and corresponding placental tissues were collected on the embryonic day 18 (E18), E20, and E22. Neonatal lung tissues were also harvested on postnatal day 1 (P1), P3, and P7. Lung sections and placental tissues were stained with hematoxylin and eosin for histological examination. Reverse transcription quantitative polymerase chain reaction (RT-PCR) analysis was performed to test mRNA expression for TLR4, myeloid differentiation 88 (MyD88) and IL-1beta, while immunohistochemistry was used to evaluate TLR4 and MyD88 expression in lung tissues. All data were analyzed with one-way analysis of variance (ANOVA) and q test. RESULTS: (1) Placental hematoxylin-eosin staining showed a great number of neutrophils infiltration, obvious interstitial hyperplasia and narrow capillaries in placental tissues in the LPS group which indicated that intrauterine infection occurred. However, there were no obvious inflammatory cells in the control group. (2) On E18, E20 and E22, the lung of LPS group showed no obvious pathological changes, and there were no apparent neutrophils infiltrated in alveoli, then some structural changes appeared. On P7, we found that the number of alveoli decreased, space of alveoli was larger than ever, septa thickened, but without significant constructive disorder. (3) In the LPS group, the TLR4, MyD88 and IL 1beta mRNA levels increased compared with control group, higher than control group at E20 and E22 (P < 0.05), and peaked at E22. Then the expression levels of these substances decreased slowly. (4) The result of immunohistochemistry showed that in lung tissues of the two groups at E18, there was no remarkable positive staining of TLR4 and MyD88, which mainly expressed in cytoplasm of bronchiole and alveolar epithelial cells, then positive cells increased slowly. CONCLUSION: (1) For perinatal rat lungs, intrauterine LPS infusion can induce an increased expression levels of TLR4 and MyD88 to a certain extent, which then returned to normal level gradually. At the same time, lung tissues showed a mild pathological change and inflammatory reaction. We propose that innate immune system of fetal lungs controls the magnitude of the LPS-induced cytokine response during the perinatal period. (2) The findings confirmed that LPS-activated signaling transduction pathway was the MyD88-dependent pathway. PMID- 20021789 TI - [Management of screening for neonatal diseases]. PMID- 20021790 TI - [Progresses in studies on hypoglycemic brain damage in neonates]. PMID- 20021791 TI - [Advances in the gene expression profile analysis and its application in the diagnosis and treatment of childhood acute leukemia]. PMID- 20021792 TI - [Clinical characteristics and follow-up of 12 cases with severe chronic active Epstein-Barr virus infection]. AB - OBJECTIVE: There are two different types of chronic active Epstein-Barr virus (CAEBV) infection: chronic EBV (CEBV) having persistent infectious mononucleosis (IM)-like illness with relatively good prognosis, and severe CAEBV (SCAEBV)infection that has rather severe manifestations and generally poor prognosis with many life-threatening complications, such as EBV-associated malignant lymphoma and hemophagocytic syndrome (HPS). The purpose of this study was to clarify the clinical and prognostic characteristics in 12 cases with SCAEBV infection. METHOD: Data of 12 cases with SCAEBV infection were analyzed retrospectively, which included clinical and auxiliary examination, pathological data, especially EB virus (EBV)-antibodies and DNA in peripheral blood mononuclear cells (PBMC) and infected tissue, and follow-up information. RESULT: Of the 12 cases, 7 were male and 5 were female. The age at the onset of diseases ranged from 35 months to 14 years (median, 11 years). The major manifestations were fever (100%), splenomegaly (91.7%), hepatomegaly (83.3%), lymphadenopathy (75.0%), and others, including skin rash, development retardation, jaundice, ascites, pulmonary hypertension, oral ulcer, cholecystitis and pleural effusion. The abnormalities of auxiliary examination were as follows: elevated LDH level (91.7%), liver dysfunction (83.3%), anemia (75.0%), leukopenia (58.3%), neutropenia (50.0%), thrombocytopenia (25.0%) and abnormal chest X-ray findings. At the time of onset, 58.3% of the patients had an IM-like illness. In all of the 12 cases, EBV serologic tests revealed high IgG antibody levels against EB viral capsid antigen (VCA). The patients often had positive IgM and IgA antibodies against VCA (33.3% and 66.7%) as well. Elevated IgG antibody level to early antigen (EA) (100.0%), occasionally positive IgA antibody (40.0%) were also seen. The mean load of EBV-DNA detected by real-time polymerase chain reaction (PCR) in the PBMC was (8.12 x 10(6), median)copies/ml. Four of 12 cases presented a poor clinical course, two of whom died from EBV-associated HPS, 1 from severe multiple pathogens infection, and 1 from multiple organ failure. In addition, 1 case developed Hodgkin's T cell lymphoma and another case showed hepatopulmonary syndrome in 2 years after splenectomy. CONCLUSIONS: The clinical feature of SCAEBV infection varied exceedingly. EBV-DNA load in PBMC of SCAEBV infected patients was significantly increased. More attention should be paid to the disease because of its severe complications, poor prognosis and high mortality. PMID- 20021794 TI - [A compound heterozygosity mutation in the interleukin-7 receptor-alpha gene resulted in severe combined immunodeficiency in a Chinese patient]. AB - OBJECTIVE: Mutation in the interleukin-7 receptor-alpha (IL-7R alpha) chain causes a rare type of severe combined immunodeficiency (SCID) with presence of NK cells in the peripheral blood. Here we report the molecular and clinical characterization of a compound heterozygosity mutation in the interleukin-7 receptor-alpha gene that resulted in SCID in a patient firstly from China. METHOD: A 5 month-old male patient and his parents were enrolled in this study. Since 15 days of age, the patient had had recurrent fever, persistent cough and diarrhea. He was in poor general condition with pyorrhea and ulceration of the BCG scar. His brother died of severe infection at 4 months of age. He was initially diagnosed as SCID according to clinical manifestation and immunological analysis. A panel of SCID candidate genes including IL-2RG, RAG1/RAG2 and IL-7R alpha of patient and his parents were amplified by polymerase chain reaction (PCR) from genomic DNA. Reverse transcription polymerase chain reaction (RT-PCR) was used to amplify the IL-7R alpha transcripts. Sequencing was performed directly on the PCR products forward and reversely. RESULT: The serum immunoglobulin (Ig) profile was IgG 6867 mg/L (normal range, 3050 - 8870 mg/L); IgM 206 mg/L and IgA 249 mg/L, IgE 2.3 IU/ml (normal range < 150 IU/ml). The patient was treated with IVIG previously. There were no T-cells but increased percentage of B-cells (58%) and NK cells (42%) in the peripheral blood was found. Needle biopsies from enlarged axillary lymph node was identified positive for Mycobacterium bovis under microscope and by culture. The patient had a compound heterozygosity mutation in the IL-7R alpha gene:on one allele, there was a splice junction mutation in intron 4 (intron 4(+1)G > A), for which his father was a carrier; whereas on the other allele, a nonsense mutation at position 638 in exon 5 with a premature stop codon (638 C > T, R206X) was identified, for which his mother was a carrier. The splice-junction mutation in intron 4 of IL-7R alpha was firstly reported. The IL-7R alpha mRNA expression of the patient was remarkably reduced whereas the parents had relatively normal IL-7R alpha mRNA expression. IL 7R alpha cDNA of the patient was amplified by nested PCR. The PCR products were purified, cloned with a TA Cloning Kit and sequenced directly. A 64 bp deletion was found in exon 4 of IL-7R alpha. No mutation was found in IL-2RG and RAG1/RAG2 of the patient and his parents. CONCLUSION: This is the first case with a compound heterozygosity mutation in the IL-7R receptor alpha gene and T-B+NK+ phenotype from China. Intron 4(+1)G > A was a novel mutation. PMID- 20021793 TI - [Clinical characteristics and treatment outcome of 36 cases with non-Hodgkin's lymphoma arising from mediastinum in children]. AB - OBJECTIVE: Non-Hodgkin's lymphoma (NHL) presenting as mediastinal mass is usually progressive and may cause severe respiratory distress and death. This study aimed to summarize the clinical features and prognosis of NHL arising from mediastinum. METHODS: Totally 36 patients with NHL arising from mediastinum reported herein were diagnosed between 1999 and 2007. Their clinical characteristics, pathologic classification, diagnosis, outcome of different treatment protocol were retrospectively analyzed. Of these 36 patients, 25 were male, 11 were female (2.2:1). The mean age was 7.9 (range 1 - 12) years. Diagnosis was established on pathology that was achieved by mediastinal mass or peripheral lymph nodes biopsy, while some were diagnosed based on bone marrow or pleural effusion cytology study and immunophenotyping. For staging, the St. Jude system was applied. Patients received T-NHL-CCCG97, T-NHL-2002 or B-NHL-2001 protocol according to morphology and immunophenotyping. Patients who experienced superior vena cava syndrome (SVCS) and/or superior mediastinum syndrome (SMS) received induction chemotherapy with cyclophosphamide (C), vincristine (O) and prednisone (P) for one week. RESULTS: Twenty-seven cases experienced mediastinal mass or peripheral lympho nodes biopsy and were diagnosed by histopathology and immunohistochemistry. Of them, 24 were lymphoblastic lymphoma and 3 were anaplastic large cell lymphoma. Nine patients were diagnosed by cytological study of bone marrow aspiration or pleural fluid. All the 36 cases were T-cell type. Twenty-four cases were in stage III, 12 in stage IV. Twenty-four patients had urgent situation of SVCS and airway obstruction, 22 patients reached good response after emergency management including COP induction chemotherapy and pleural effusion suction. Twenty-nine cases achieved complete remission (CR) while in 6 patients the disease relapsed. Thirteen patients died from disease progression, relapse or severe infection during chemotherapy. The Kaplan-Meier estimate of 5-year progression-free survival (PFS) was 61% +/- 8% (median follow up 35 months) for these 36 patients. CONCLUSION: Establishment of a diagnosis as soon as possible was important to reduce the mortality and improve long term survival of patients. Induction chemotherapy for emergency situation was efficacious. The regimen of T-NHL CCCG97, T-NHL-2002, and B-NHL-2001 for NHL arising from mediastinum based on pathological classification is feasible. PMID- 20021795 TI - [Mutation of NKX2-5 gene in patients with atrial septal defect]. AB - OBJECTIVE: The purpose of this investigation was to identify the novel genetic mutations in patients with a congenital atrial septal defect (ASD). METHODS: Clinical data and blood specimens from a total of 12 unrelated ASD pedigrees and a cohort of 168 unrelated subjects with sporadic ASD were collected and evaluated in contrast to 200 healthy individuals. The whole exons and partial flanking introns of NKX2-5 gene were amplified by polymerase chain reaction and sequenced using the di-deoxynucleotide chain termination approach. The acquired sequences were aligned with those publicized in GenBank by the aid of programme BLAST to identify the sequence variations. The software ClustalW was applied for analysis of the conservative of the altered amino acids. RESULTS: A novel heterozygous mutation of NKX2-5 gene, i.e., a substitution of thymine for cytosine at nucleotide 536, predicting the conversion of serine into phenylalanine at amino acid residue 179, was identified initially in a proband. The same missense mutation was thereafter detected in other 3 affected members of the identical family but neither in the healthy members of the kindred nor in the 200 normal controls. A cross-species alignment of the protein sequences encoded by NKX2-5 gene displayed that the mutated amino acid was highly conserved evolutionarily. No mutation of NKX2-5 gene was observed in the other 11 ASD pedigrees or in 168 patients with sporadic ASD. Additionally, a common synonymous single nucleotide polymorphism, a transition of adenine (A) into guanine (G) at nucleotide 63, was found in NKX2-5 gene. However, there were no significant differences in the prevalence of alleles A and G between ASD patients and healthy controls (chi(2) = 2.8641, P = 0.0906). CONCLUSION: A novel mutation of NKX2-5 gene identified in an ASD family suggests that mutated NKX2-5 gene is responsible for familial ASD. PMID- 20021796 TI - [Efficacy of biofeedback therapy in children with functional constipation]. AB - OBJECTIVE: To evaluate the benefits of biofeedback therapy in children with functional constipation caused by pelvic floor dysfunction (PFD). METHOD: Anorectal manometry (PC Polygraf HR, Medtronic) was performed in children with functional constipation according to the diagnostic criteria of Rome III. Among them, 47 cases with PFD were trained with biofeedback therapy (PC Polygraf HR, Medtronic) according to the degree of cooperation. They received the treatment twice every week, and those in whom the therapy was performed at least three times were enrolled to evaluate the efficacy in this study. RESULT: A total of 27 cases (20 male and 7 female) in whom the procedure was performed at least three times were eligible for inclusion into this study. The mean age of them was (6.7 + or - 2.2) years (range 4 to 12 years), and the mean duration of symptoms was (3.0 + or - 2.3) years with a range of 6 months to 8 years. Among them, 16 cases received the biofeedback training more than 5 times, while 8 cases more than 7 times. The rectal maximum contraction pressure during defecation was increased significantly with the number of biofeedback training before treatment and after 3, 5, and 7 times of treatment was (36.2 + or - 10.4), (45.1 + or - 9.5), (47.6 + or - 9.1), and (47.8 + or - 8.8) mm Hg (1 mm Hg = 0.133 kPa) respectively (P < 0.01). The potential of electromyography of external anal sphincter decreased progressively, but had not reached significance. Follow-up continued for 3 to 12 months by telephone, the clinical response to biofeedback treatment was evaluated as excellent (complete resolution of constipation), good (improvement of constipation), partial or poor (no improvement of constipation). The response was excellent in 13 cases (48.1%), good in 8 (29.6%), partial in 3 (11.1%), and 2 (7.5%) cases had no improvement, and 1 case was lost to follow-up, and the rate of success was 88.9% (24/27). CONCLUSION: Biofeedback therapy is a safe and effective treatment option for functional constipation due to PFD in children probably by increasing the rectal maximum contraction pressure during defecation. PMID- 20021797 TI - [Radiofrequency catheter ablation of arrhythmias in pediatric patients guided by three-dimensional mapping system]. AB - OBJECTIVE: To explore safety, indications and advantages of mapping and ablation of arrhythmia in children guided by Carto and Ensite system. METHODS: Guided by Carto system, radiofrequency catheter ablation (RFCA) was performed on 8 pediatric patients with tachycardia whose mean age was (6.2 + or - 1.7) years, mean weight was (18.0 + or - 2.0) kg. Guided by Ensite system, RFCA was performed on 10 pediatric patients with arrhythmia, 8 of them were ablated guided by Ensite Array system: 6 cases with premature ventricular contractions (PVCs), 2 cases with right atrial tachycardia, their mean age was (11.3 + or - 1.2) years, and mean weight (40.0 + or - 5.0) kg. The other two cases with W-P-W syndrome were ablated guided by Ensite Navx system. RESULT: Guided by Carto system, 8 cases were successfully mapped and ablated: 6 cases had incision atrial tachycardia, 1 case had left atrial tachycardia and 1 case had right atrial tachycardia. In 1 case with incision atrial tachycardia the condition recurred after 3 months, and was ablated again successfully. Guided by Ensite Array system, 6 cases with PVCs (in 2 originating from the right ventricular inflow tract and in 4 originating from the right ventricular outflow tract) and 2 cases with right atrial tachycardia were successfully mapped and ablated, PVCs of the first 6 cases were reduced from (32 333 + or - 4509) 24 h to (0-4)/24 h after ablation. In 1 case with automatic atrial tachycardia, mapping could not be done by Ensite Array system, because P wave could not be identified from T wave. Single bolus of adenosine 20 mg was given within 30 s to let ventricles stop for 2 s (cardio ventricular pacing standby) until T wave vanished, mapping and ablation were operated again successfully, but another atrial tachycardia occurred 1 day later. Guided by Ensite Navx system, 2 cases with W-P-W syndrome were successfully ablated, operation under X-rays lasted for 8 and 10 min. In none of the 9 patients the disease recurred after follow-up for 6 months. CONCLUSION: Carto system is suitable for mapping and ablation in pediatric patients with continuous tachycardia, especially with incision atrial tachycardia; Ensite Array system fits children older than 10 years with right heart discontinuous arrhythmia; and Ensite NavX system can set up model and display endocardial anatomic structure quickly. Compared with two-dimensional mapping system, the three-dimensional mapping system (Carto and Ensite) can display the origin of arrhythmia and activation sequence clearly, decrease difficulty of operation efficiently and diminish operation time under X-ray. PMID- 20021798 TI - [Retrospective analysis of 76 children with acute promyelocytic leukemia]. AB - OBJECTIVE: There are very limited data on childhood acute promyelocytic leukemia (APL), especially childhood APL treated with arsenic trioxide (As(2)O(3)). METHOD: Between January 1999 and December 2007, 76 children (< 18 years) with newly diagnosed APL were included. RESULTS: Six cases failed to achieve hematological complete remission (CR) due to early death. Three groups of patients were identified according to the different induction regimen. Forty-four patients were given all-trans retinoic acid (ATRA) alone (group1, G1), 7 patients were given As(2)O(3) alone (group 2, G2), 19 patients (group 3, G3) were treated with combination of ATRA and As(2)O(3). The CR rate of G1 was 100%. The overall CR rate of G2 + G3 was 100%. Five children underwent hematologic relapse and 2 developed molecular relapse. The 5-year cumulative incidence of relapse was 13.8%, whereas event-free (EFS), disease-free (DFS) and overall survival rates were 79.5%, 86.3% and 90.5%, respectively. The 5-year estimate of EFS and DFS between G1 and G2 + G3 had no significant difference. CONCLUSION: As(2)O(3) is an effective and well tolerable therapy for children with APL and it may be used in those who not only cannot bear side effects of ATRA but also the newly diagnosed and relapsed APL. PMID- 20021799 TI - [Progresses in studies on supplementation of vitamin D]. PMID- 20021801 TI - [Atrioventricular nodal reentrant supraventricular tachycardia of slow and rapid pathways position in a child with atrial septal defect of primary and secondary opening reversed by radiofrequency catheter ablation]. PMID- 20021802 TI - [Professor CHEN Cui-zhen: in pursuit of her dream to give sick children the best available care for her whole life]. PMID- 20021803 TI - [Update on pediatric flexible bronchoscopy in China]. PMID- 20021804 TI - [Airway obstruction caused by large blood vessel anomalies: assessment by flexible bronchoscopy]. AB - OBJECTIVE: To evaluate the diagnostic value and safety of flexible bronchoscopy in congenital great vessel diseases complicated with airway compression. METHOD: The medical records of patients with great vessels abnormalities who were admitted to the neonatal intensive care unit (NICU) from October 2005 to June 2009 were retrospectively reviewed; 34 cases were diagnosed as airway compression by flexible bronchoscopy, 10 cases as vascular ring, 24 cases as aortal arch obstruction. The age of the patients was 6 d - 11 m, body weight 2.2 - 8.7 kg [(4.6 +/- 1.4) kg]. Recorded airway abnormalities detected by bronchoscopy and CT, cardiac vascular defects and airway compression were consistent with the findings on operation. The relation between the airway compression and cardiac vascular abnormalities, treatment of the airway compression and outcome were analysed. RESULT: Bronchoscopic assessment was successfully performed in NICU or operating room for all the patients. (1) Initial presentation of the 34 cases were tachypnea, stridor, refractory lung infection and prolonged mechanical ventilation. (2) Extrinsic compression was found in all the 10 cases with vascular ring by bronchoscopy initially which indicated vascular ring, airway compression was mainly of lower part of trachea. Diagnosis of 9 cases was consistent with CT diagnosis and in 1 case the diagnosis was confirmed by surgery; among these cases, 7 had congenital tracheal stenosis. (3) In the 24 cases with aortic obstructive lesion, 5 were detected to have tracheal stenosis by CT before correction of vascular abnormality, among whom one case was indicated to have tracheal stenosis by bronchoscopy, the other 19 cases were found with airway compression by bronchoscopy during or after vascular correction. Among the 24 cases, 21 had left main bronchial stenosis, 2 had congenital tracheal stenosis. Airway compression diagnosed by bronchoscopy agreed with the findings of CT. Two cases developed transient decrease of oxygen saturation, 5 cases developed transient tachycardia. CONCLUSION: Flexible bronchoscopy plays an important role in assessment of the airway compression complicated with great vessel abnormalities. Bronchoscopy is an accurate, convenient, safe and rapid way for airway assessment, but further examination of the peripheral structure and vascular malformation need combined examination with CT. PMID- 20021805 TI - [Diagnosis and treatment of invasive pulmonary aspergillosis in 21 children with non-hematologic diseases]. AB - OBJECTIVE: To explore diagnosis and treatments of invasive pulmonary aspergillosis (IPA) in children with non-hematologic diseases. METHOD: Twenty one patients without hematological malignancy were diagnosed with proven or possible IPA from July 2002 to June 2008. The risk factors, clinical manifestations, chest radiographic findings, microbiological and histopathological evidence, diagnostic procedures, treatment and prognosis were retrospectively reviewed. RESULT: Five children had proven IPA, and 16 patients had possible IPA. Thirteen children were classified as having acute invasive pulmonary aspergillosis (AIPA), eight children as having chronic necrotizing pulmonary aspergillosis (CNPA). Definitive diagnosis of primary immunodeficiency (PID) was made in 6 children (4 with chronic granulomatous disease, 2 with cellular immunodeficiency); three children were suspected of having PID. Corticosteroids and multiple broad-spectrum antibiotics had been administered in 5 patients (3 of these 5 patients also had invasive mechanical ventilation). Two children had underlying pulmonary disease. Three patients had unknown risk factors. Among these three patients, two had history of environmental exposure. Fever and cough were present in all the children. Fine rales were found in nineteen children. Six children had hepatosplenomegaly. The common roentgenographic feature of AIPA in 13 patients was nodular or mass-like consolidation with multiple cavity. "air-crescent" was seen in 10 of patients with AIPA. Lobar consolidation with cavity and adjacent pleural thickening was found in all children with CNPA. The positive rate of sputum and/or BALF culture in AIPA and CNPA were 72.1% and 22.4%, respectively. A large number of septate hyphae on wet smear were found in all of the children whose sputum and/or BALF culture were positive. Lung biopsy was performed in 3 children with CNPA, and necrosis, granulomatous inflammation, as well as septate, branching hyphae were observed on histopathologic examination. Fifteen children were treated with anti-fungal therapy (amphotericin B, voriconazole, itraconazole and caspofungin used alone or in combination), symptoms and lung lesions resolved in 12 children. Three children died. Six children did not receive anti-fungal therapy and died. The side effects of amphotericin B include chill, fever, hypokalemia and transient increase in BUN, none of which needed discontinuation of the antifungal therapy. Children had a good tolerance to fluconazole and caspofungin, there were no apparent side effects. CONCLUSION: Most of the children without hematologic diseases who suffered from invasive pulmonary aspergillosis had risk factors or exposure history. Roentgenographic findings were relatively characteristic for invasive pulmonary aspergillosis. Risk factors and roentgenographic findings were clues to consider clinically invasive pulmonary aspergillosis. Sputum culture was the key point to clinical diagnosis. The patients in whom the antifungal therapy was initiated early had a good outcome. PMID- 20021806 TI - [Effect of vitamin D supplementation in early life on the expression of interleukin-10 and intercellular adhesion molecule-1 in rat asthma model]. AB - OBJECTIVE: To explore the effect and mechanism of vitamin D supplementation in early life on rat asthma model. METHOD: Thirty two sex-mature, female Wistar rats were randomly divided into a control group (n = 8), a low dose group (n = 8), a medium dose group (n = 8) and a high dose group (n = 8). From the seventh day of pregnancy on, the rats in each group were given different doses of vitamin D by intragastric administration, until the offspring rats were 21 days old. The rats in the control group were fed with DMSO-PBS. After the offsprings were weaned, 8 rats were randomly selected from each group. The number of male and female rats was equal. The rats were sensitized to ovalbumin (OVA) and challenged with aerosol OVA to establish the asthma model. The lung tissues were examined for pathologic changes after HE staining. ELISA was used to determine the concentrations of IL-10 in serum and BALF. Immunohistochemical staining methods were used to measure the expression of intercellular adhesion molecule-1 (ICAM-1) in lung tissues. RESULT: (1) Pathologic changes of lung tissues: compared with the control group, light microscope (LM) showed that eosinophil cells infiltration and the airway inflammation decreased in the low dose and medium dose groups, but increased in the high dose group. (2) The concentrations of IL 10 in serum and BALF: In serum, compared with the control group [(18.7 +/- 4.7) pg/ml], the concentrations of IL-10 in the low dose group [(30.2 +/- 2.8) pg/ml, P < 0.05] and the medium dose group [(51.5 +/- 6.6) pg/ml, P < 0.05] were significantly increased. And the IL-10 level of medium dose group was higher than that of the low dose group (P < 0.05). In BALF, compared with the control group [(59.1 +/- 14.4) pg/ml], the concentrations of IL-10 in the medium dose group [(90.0 +/- 14.3) pg/ml, P < 0.05] was significantly increased. There were no significant changes in the low dose group [(58.1 +/- 3.4) pg/ml, P > 0.05], whereas in the high dose group [(45.3 +/- 6.5) pg/ml, P < 0.05] the level significantly decreased. (2) The expression of ICAM-1 in lung tissues: compared with the control group, there were no significant changes in the low dose group (P > 0.05). The expression of ICAM-1 was significantly decreased in the medium dose group (P < 0.05). In the high dose group, the expression of ICAM-1 was significantly increased (P > 0.05). CONCLUSION: Adequate intervention with 1,25(OH)2D3 in the early life could alleviate the inflammation in the lung tissues, reduces eosinophil cell infiltration in rat asthma model. However, overdose might play a detrimental role. Its mechanism may be associated with the effect of 1,25(OH)2D3 on IL-10 secretion and the expression of ICAM-1. PMID- 20021807 TI - [Guide to pediatric bronchoscopy (2009 edition)]. PMID- 20021808 TI - [Clinical value of wireless capsule endoscopy in diagnosis of small bowel disease in children]. AB - OBJECTIVE: The pathological change of small bowel is difficult to examine because it is anatomically unique. The development of wireless capsule endoscopy provides an unique opportunity to visualize the entire small bowel in a minimally invasive manner. The aim of this study was to assess the safety and clinical value of wireless capsule endoscopy in children. METHODS: During the last 4 years (June, 2004-June, 2008), 46 times of wireless capsule endoscopy were performed in 43 patients with suspected small bowel disease, including obscure gastrointestinal bleeding (n = 11), recurrent abdominal pain (n = 20), chronic diarrhea (n = 9), protein losing enteropathy (n = 2), recurrent vomiting (n = 1). Of the 43 cases, 28 were male and 15 were female, the age ranged from 6 to 18 years, 8 of these cases were < 10 years old. The weight of the patients ranged between 15 kg and 60 kg. The average time of capsule passing through the stomach and the small intestine, the tolerance to and complication of wireless capsule endoscopy in patients, the image quality of capsule endoscopy, and the cleanliness of small intestine after fasting for 8 hours were observed and recorded. RESULT: All the patients could easily swallow the capsule and had good tolerance. The overall success rate was 94% (43/46). The median time of capsule passing through the stomach and small intestine was 73 min (range, 3 - 600 min) and 246 min (range, 73 - 413 min), respectively. The diagnostic yield of pathological change in small intestine was 90% (37/41), and the diagnostic accordance rate was 84% (31/37). Based on the wireless capsule endoscopy, diagnostic findings included Crohn's diseases (15), lymph follicular hyperplasia (4), nonspecific enteritis (4), vascular malformations (3), small bowel tumour (2), primary intestinal lymphangiectasia (2), gastrointestinal motility disorders (2), Meckel's diverticulum (1), angioma (1), small intestinal worm disease (1), duodenal ulcer (1), and polyposis syndromes (1). The capsule of 1 patient remained in the stomach. The cleanliness of small intestine after 8 hours fasting was good. And the capsule endoscopy can show high quality small intestine image. CONCLUSION: Wireless capsule endoscopy is a noninvasive, safe and useful tool for the investigation of the small intestine in children, especially for obscure gastrointestinal bleeding and Crohn's disease. PMID- 20021809 TI - [Polyneuro-electrophysiological studies of myoclonus in children]. AB - OBJECTIVE: To explore the clinical and neuroelectrophysiological characteristics of myoclonus of different origins in children. METHOD: Thirty-two children with myoclonic seizure were analyzed by video electroencephalogram-electromyogram (VEEG-EMG) polygraphic recordings, jerk-locked back averaging (JLA) and short latency somatosensory evoked potential (SSEP). They were classified into cortical myoclonus (CM), subcortical myoclonus (SCM), and unidentified group according to their generating locations, and also were classified into epileptic and non epileptic myoclonus based on their different properties. RESULT: The 32 patients included 14 with CM, 14 with SCM and 4 with unidentified origin. (1) CM group: the myoclonic patients presented as focal and/or multifocal seizures in 11 cases and as generalized in another 3 patients besides focal myoclonus. Arrhythmic jerks were shown completely in 11 cases and rhythmic seizures were concomitant in another 3 patients. Myoclonus sensitivity to sensory stimulus was observed in 10 patients. The durations of EMG burst were 10-52 ms. Background EEGs were presented normal in 4 patients and slowing in 10 patients. The epileptiform discharges in interictal EEG were variable. The ictal EEG showed epileptic discharges with each clinical jerk in 9 cases but only with some jerks in 4 patients. Another one had no any EEG abnormality in each jerk. The myoclonus related spikes were disclosed in 13 cases by JLA. Of the 10 cases who underwent SSEP, giant SSEPs were seen in 3 cases including the one with normal EEG and JLA analyses. (2) SCM group: myoclonus was presented as generalized in 8 cases and as focal in 6 cases. All the patients showed arrhythmic jerks and 14 cases were not sensitive to stimulus. The durations of EMG burst were from 60 ms to 400 ms. Normal background EEGs were presented in 6 patients and slowing in 8 patients. The interictal EEG showed no consistent abnormality. Epileptic discharges associated with myoclonus seizures were not found in any of 9 patients but were observed with some seizure changes in 5 cases. There was no myoclonus-related spike by JLA in this group. SSEPs were normal in all patients. (3) The group with unidentified origin: the durations of EMG were from 60 ms to 400 ms, and their EEG and SSEP recordings were normal. In addition, 32 patients could be classified as epileptic myoclonus in 14 cases and nonepileptic myoclonus in 18 cases by the polyneurophysiological tests. CONCLUSION: (1) It is not reliable to identify myoclonus seizures and their clinical properties depending on their interictal and ictal EEGs only. (2) Polyneuroelectrophysiological tests, including EEG-EMG, JLA, and SSEP, seem to be valuable and useful to identify the generating locations and properties for different myoclonus in children. PMID- 20021810 TI - [Screening and risk factors analysis of retinopathy of prematurity]. AB - OBJECTIVE: To analyze the risk factors of retinopathy of prematurity (ROP) and provide evidence for the rational establishment of screening standard. METHODS: The clinical data of 1675 preterm infants at gestational age < or = 36 weeks or birth weight < or = 2500 g who were admitted to the neonatal intensive care unit and had been screened in our hospital from July 2006 to May 2008 were analyzed retrospectively by univariate analysis and Logistic regression analysis. Gender, birth count, gestational age, birth weight, oxygen therapy, and mother's conditions were recorded. RESULTS: ROP was detected in 195 (11.6%) of 1675 infants, of whom 35 infants (2.1%) had type 1 or threshold ROP. The lower the birth weight, the smaller the gestational age and the longer the time of oxygen therapy were, the higher the incidence of ROP was. For the infants whose birth weight was < or = 1200 g, 1201 - 1500 g, 1501 - 2000 g, 2001 - 2500 g, the incidence of ROP was 73.2%, 30.4%, 8.0%, and 1.1%; for those at gestational age < or = 30 weeks, 30(+1)-32 weeks, 32(+1)-34 weeks, 34(+1)-36 weeks, the incidence of ROP was 67.6%, 16.9%, 3.9%, and 1.0%; for the infants underwent oxygen therapy for 0 d, -3 d, -5 d, -8 d, > 8 d, the incidence of ROP was 1.5%, 3.3%, 9.6%, 23.2% and 38.8%;in the infants who inhaled oxygen at concentrations of 0.40, 0.60, -0.80 and > 0.80, the incidence of ROP was 11.8%, 18.1%, 26.8%, and 52.6%, respectively. Logistic regression analysis indicated that low birth weight, small gestational age, asphyxia, apnea, oxygen therapy were the high risk factors of ROP (the odds ratio was 0.957, 1.052, 1.186, 5.314, and 1.881). CONCLUSIONS: Low birth weight, small gestational age, asphyxia, apnea, and oxygen therapy were the high risk factors of ROP. It is recommended that all preterm infants with high risk factors should be screened. PMID- 20021811 TI - [Four new cases with WT1 gene mutations in Chinese patients with Wilms' tumor]. AB - OBJECTIVE: Wilms' tumor (WT) is the most common malignant renal tumor in childhood. The WT1 gene located at 11p13 was identified in 1990 as a tumor suppressor gene important in the development in WT. The WT1 gene consists of 10 exons, with exons 1 to 6 encoding an N-terminal proline- and glutamine-rich transactivational domain, and exons 7 to 10 encoding a C-terminal zinc-finger domain involved in DNA binding. In China we know little about the frequency and genotype of WT1 mutations in Chinese WT patients. This study aimed to determine the frequency and genotype of WT1 mutations in children with nonsyndromic WT in China. METHODS: We collected peripheral blood of WT patients treated in Beijing Children's Hospital. Genomic DNA of 54 WT patients was isolated from blood samples. All coding WT1 exons and their flanking intronic sequences were amplified by PCR method. The amplified PCR products from all individuals were then subjected to automatic DNA sequencing. RESULTS: Four different constitutional WT1 mutations were identified in four children. Three mutations are predicted to produce truncated protein. One mutation is missense. Of the four mutations, three had not been reported before. Patient 1 had a 1006 A > T transition in exon 7, which caused (336)Lys to become a stop codon (K336X). DNA sequence analyses in patient 2 indicated the point mutations in exon 9 which was a 1168 C > T substitution and caused (390)Arg to become a stop codon (R390X). It indicated a point mutations in exon 6 in patient 3 which was a 814 G > T substitution and resulted in (272)Glu to become a stop codon (E272X). In patient 4 there was a homozygous mutation in exon 10. The mutation was a 1228 A > G substitution and resulted in (410)Ser to become a Gly codon (S410G). CONCLUSION: Constitutional WT1 mutations occur at a low frequency (7.4%) in Chinese patients with Wilms' Tumor. It is similar to the results of overseas study. Four WT1 gene mutations were confirmed, three were nonsense, one was missense. PMID- 20021812 TI - [Increased apoptosis after intra-amniotic endotoxin priming plus hyperoxic exposure in lungs of preterm newborn rats]. AB - OBJECTIVE: Apoptosis has been shown to be involved in lung remodeling in both rat lung and human fetal lung explants. Mounting evidence suggests the pro- and anti apoptotic members of the Bcl-2 family exert most of their function at the mitochondrial level and play pivotal roles in the process of apoptosis. Molecules such as Bax (death agonist) and Bcl-2 (death antagonist) act in competition, and their relative abundance and dimerization can determine cell death or cell survival. It is not known whether apoptosis is in part responsible for the impaired lung growth found in preterm developing lung exposed to intra-amniotic endotoxin priming plus hyperoxia, and imbalance of Bax/Bcl-2 contributes to it. The purpose of this study was to investigate the expression apoptosis index (AI), Bax and Bcl-2 in premature lungs of rats with intra-amniotic endotoxin priming and/or exposed to 60% O(2) and to elucidate the relationship between intrauterine inflammatory/chronic O(2) exposure and the pathogenesis of bronchopulmonary dysplasia (BPD). METHODS: Timed pregnant Sprague Dawley (SD) rats were randomly divided into two groups: lipopolysaccharide (LPS) group and saline group. LPS or saline were intra-amniotically injected into the sacs on gestational age day 15 (70% of term). Six days after intra-amniotic injection, the preterm rats were delivered and randomized to put in 60% O(2) exposure or in room air. On days 1, 7 and 14 after birth, the lungs were removed and dissected from the main bronchi for analysis. Total RNA were extracted from the right frozen lung tissues. Lung AI was measured by terminal transferase nick end labeling (TUNEL) and Bax and Bcl 2 mRNA levels were measured by semi-quantitative reverse transcription polymerase chain reaction (RT-PCR). RESULTS: (1) To quantify the proliferation of preterm lungs after intra-amniotic endotoxin priming and/or exposed to 60% O(2), proliferating cell nuclear antigen (PCNA) in the four experimental groups were measured: PCNA increased along with the lung development from d1 to d14 in both control and endotoxin alone groups. However, in endotoxin alone group, PCNA was significantly lower than that of controls from d1 to d14. In hyperoxia alone group, PCNA begun to decreased on day 7, and became significantly lower than that of the control group on day 14 (P < 0.05). In endotoxin plus hyperoxia group, PCNA was significantly lower than that of controls on days 1 and 7. (2) To quantify the apoptosis of immature lungs after intra-amniotic endotoxin priming and/or exposed to 60% O(2), AI increased along with the lung development from d 1 to d 7 in both control group and endotoxin alone group. However, in endotoxin alone group, AI was significantly higher than that of controls from d1 to d7. In hyperoxia alone group, AI begun to increased on day 7, and became significantly lower than that of the control group on day 14 (P < 0.05). In endotoxin plus hyperoxia groups, AI was higher than controls on days 1 and 7, and became significantly higher than those of the other 3 groups on day 14. (3) RT-PCR showed that mRNA of Bax decreased while Bcl-2 mRNA increased from day 1 to day 14 in endotoxin alone group, and they were significantly higher than those of controls on day 1 (P < 0.05). Conversely, Bax mRNA increased while Bcl-2 mRNA decreased from day 1 to day 7 in hyperoxia treated groups, and was significantly different from that of the control group on days 7 and 14 in both hyperoxia treated groups. CONCLUSIONS: These results suggest that intra-amniotic endotoxin presensitization and exposure to hyperoxia may inhibit proliferation of lung cells. Increased expression of Bax and Bcl-2 may be related to the evolution of lung injury and may contribute to the pathogenesis of BPD. PMID- 20021813 TI - [Stanozolol activates the cross-talk of estrogen receptor alpha-insulin-like growth factor-1 receptor-extracellular-signal regulated kinase 1/2 in the growth plate chondrocytes of estrogen-inhibited adolescent rats in vitro]. AB - OBJECTIVE: To investigate the effects and the mechanisms of stanozolol (ST) on the proliferation, maturation and differentiation of in vitro cultured growth plate chondrocyte isolated from gonadotropin releasing hormone analogue (GnRHa) treated adolescent rats, to study if ST mediates the proliferation of chondrocytes via the estrogen receptor alpha (ERalpha), androgen receptor (AR) and/or insulin-like growth factor-1 receptor (IGF-1R) and interactions of the two receptor and IGF-1R receptor signaling pathway, to investigate the mechanism of the biological effects in ST promoting bone growth/maturity at molecular level. METHOD: The rats were weaned at the end of 3 weeks and intramuscular injection of triptorelin of GnRHa preparations, qow x 2 was started. The rats were sacrificed at the end of 7 weeks, and then the tibiae growth plates were taken out with sterile procedure. The chondrocytes were obtained by two-time enzyme digestion method, and the experiments were carried out with the primary chondrocytes. Immunohistochemical staining of proliferating cell nuclear antigen (PCNA) and Western blot analysis were applied. RESULT: The results of PCNA demonstrated that stanozolol enhanced the proliferation of the chondrocytes, time-course studies showed that the proliferation were maximally stimulated by stanozolol after 2 days of incubation and decreased again after longer periods of incubation. The expression of p-ERalpha, p-IGF-1R and p-extracellular-signal regulated kinase 1/2 (ERK1/2) increased with the incubation period of ST treatment, and reached the peak value at a certain time, and then gradually decreased. The expression of p ERalpha, p-IGF-1R and p-ERK1/2 increased with the elevation of ST concentration, and reached the peak value at 10(-9) - 10(-8) mol/L, then gradually decreased. ST induced-p-ERalpha expression was partially blocked by ERalpha and mitogen activated protein kinase kinase inhibitors. ST induced-p-IGF-1R expression was partially blocked by ERalpha and IGF-1R inhibitors. ST induced-p-ERK1/2 expression was partially blocked by mitogen-activated protein kinase kinase and IGF-1R inhibitors. CONCLUSION: As an androgen derivation, ST exerts its biological effects of promoting proliferation of the long bone growth plate chondrocytes via activating the classic ERalpha receptor pathway and mitogen activated protein kinase pathway, and at the same time, by activation of IGF-1R. Both IGF-1R and ERalpha can promote "cross-talk" of two systems' receptor signal through mitogen-activated protein kinase signal pathway. PMID- 20021814 TI - [Analysis of the causes of pediatric rhino-source diseases]. AB - OBJECTIVE: To analyze clinical features of pediatric rhino-source diseases for reducing missed diagnosis and misdiagnosis. METHOD: Data of 3588 children with rhino-source diseases seen from April 2005 to May 2006 were retrospectively analyzed in this study in order to disclose the relationship of etiological factor, clinical features and diagnosis. RESULT: Among all these cases, 2090 complained of nasal discomfort including nasal obstruction, discharge, rhinalgia and epistaxis. However 1498 cases (41.76%) did not, of whom 470 cases had snoring and apnea, 332 cases of otalgia and otorrhea, 145 cases had chronic cough, 138 had headache and 92 had lower respiratory infection. CONCLUSION: A high percentage of children who suffered from pediatric rhino-source disease did not develop nasal symptoms. Pediatric rhino-source disease should be considered for patients in whom the therapeutic effect is unexpectedly poor. PMID- 20021815 TI - [Congenital intrapulmonary lymphangioma and pulmonary hemangioma in 5 infants and young children]. AB - OBJECTIVE: To analyze the clinical, imaging and pathological findings of congenital intrapulmonary lymphangioma and hemangioma in 5 infants and young children. METHOD: Data of 3 cases with congenital intrapulmonary lymphangioma and 2 cases with haemangioma were analyzed. RESULT: All the 5 cases had cough, difficulty in breathing, cyanosis of lips, and shortness of breath. Imaging study indicated cystic and space occupying changes of partial lung. In the two cases of hemangioma, the blood vessels passing through the hemangioma were observed on CT films. Histochemical studies showed that the cystic pockets from removed tissues were different in size and there was a line of flat endothelial tissue around these pockets. Immunochemical studies indicated D2-40 positive, factor VIII weak positive for lymphangioma cases; while in hemangioma cases, factor VIII was positive, D2-40 was negative or weakly positive, and both of cytokeratin was negative. CONCLUSION: It is very important to consider and identify congenital intrapulmonary lymphangioma or hemangioma when a patient has cystic pockets and space occupying change in their lung by imaging and pathological studies. PMID- 20021816 TI - [Expression of CD147 and matrix metalloproteinase-9 in children with non Hodgkin's lymphoma and its correlation with prognosis]. AB - OBJECTIVE: To investigate the expression of CD147 and matrix metalloproteinase-9 (MMP-9) in children with non-Hodgkin's lymphoma (NHL) and its correlation with clinical stage, tumor size, bone marrow invasion, immunological typing, serum lactate dehydrogenase (LDH) concentration, and prognosis. METHODS: Specimens excised from NHL patients were prepared. Expression of CD147 and MMP-9 were tested by streptavidin-biotin complex (SABC) immunohistochemistry and its correlation with clinical results were analyzed. RESULTS: The positive rate of CD147 expression was 73% (45/62), 17 cases were (-), 11 cases (+), 34 cases (++) and 21 cases (+++). The positive rate of MMP-9 expression was 81% (50/62), 12 cases were (-), 13 cases (+), 18 cases (++) and 19 cases (+++). The Spearman rank correlation analysis indicated that there was a positive correlation between CD147 and MMP-9 expressions in NHL (r(S) = 0.763, P = 0.034). Expression of CD147 was determined in relation to factors that included clinical bone marrow invasion, tumor size, LDH level as well as the clinical stage; expression of MMP 9 had a positive correlation with bone marrow invasion, tumor size and clinical phases. The 5-year survival rates (5YSR) were 78% (22/28) and 45% (15/34) in the cases whose CD147 expression was (-)-(+) and (++)-(+++), respectively, and 5YSR were 84% (21/25) and 43% (16/37) in the cases whose MMP-9 expression was (-)-(+) and (++)-(+++) respectively, the difference was significant. Cox multivariate analysis showed that both CD147 and MMP-9 were important prognostic factors. CONCLUSION: The increased expression of CD147 and/or MMP-9 correlates with a poor clinical outcome in patients with NHL. PMID- 20021817 TI - [Genetic mutation analysis of a kindred with a patient suffering from 17 alpha hydroxylase/17, 20-lyase deficiency]. AB - OBJECTIVE: To detect CYP17A1 gene mutation in a patient with 17 alpha hydroxylase/17, 20-lyase deficiency and her family members. METHOD: Genomic DNA was extracted from the blood of the patient, her parents and twin sister. The 8 exons of CYP17A1 gene were amplified with polymerase chain reaction (PCR) and screened for mutations by sequencing. RESULT: The analysis revealed that the patient was a compound heterozygote carrying two different inherited point mutations on CYP17A1 gene. They were nt186delC on exon 1 and nt1085G > A on exon 6. This type of mutation could induce 17OHD because of complete loss of 17 alpha hydroxylase activities. And her parents and the twin sister were carriers on CYP17A1 gene. In addition, the mutation nt186delC was a novel point mutation and it was not discovered in normal children. CONCLUSION: A new compound heterozygote carrying two different inherited point mutations on CYP17A1 gene was found, and her parents and twin sister were carriers. This is probably the first report in the world of a twin sisters of whom one is a patient with 17OHD and the other is a carrier of CYP17A1 gene mutation. PMID- 20021818 TI - [A case of Xp21 contiguous gene deletion syndrome: clinical and genetic study]. PMID- 20021819 TI - [Progress in studies on Panayiotopoulos syndrome]. PMID- 20021820 TI - [Acute lymphoblastic leukemia in adolescents]. PMID- 20021821 TI - [Inhibitory effect of knocking down microRNA-221 and microRNA-222 on glioma cell growth in vitro and in vivo]. AB - OBJECTIVE: To study the inhibitory effect of knocking down microRNA(miR)-221 and miR-222 on human glioma cell growth and its possible mechanism. METHODS: miRNA 221/222 antisense oligonucleotides (antisense miR221/222) were transfected into human glioma U251 cells by lipofectamine. Northern blot analysis was conducted to detect the mRNA expression of miR-221/222 in the control and transfected cell groups. The proliferation activity of cells was determined by MTT assay. Cell invasion ability was examined by transwell assay, and cell cycle kinetics and apoptosis were detected with flow cytometry. The expression of relevant proteins was analyzed by Western blotting. The therapeutic efficacy of antisense miR221/222 on the growth of xenograft tumors in nude mice were also observed. RESULTS: In the antisense miR-221/222-transfected cells, the expression of miR 221/222 was significantly reduced; the cell invasion ability was suppressed, cell cycle was blocked at G(0)/G(1) phase, and apoptotic cells were increased. The growth of xenograft tumors treated with antisense miR-221/222 was also inhibited. In antisense miR-221/222 treated tumor cells, the expression of bcl-2 was down regulated while connexin43, p27, PUMA, caspase-3, PTEN, TIMP3 and Bax up regulated, and p53 expression not changed. CONCLUSION: There is a significant inhibitory effect of antisense miR-221/222 on the growth of human glioma U251 cells. miR-221/222 may be considered as a candidate target for gene therapy of human gliomas. PMID- 20021822 TI - [MT1-MMP up-regulates VEGF expression in human breast carcinoma MCF-7 cells and induces tumor angiogenesis]. AB - OBJECTIVE: To study the function of MT1-MMP in tumor angiogenesis and elucidate the possible way of action that MT1-MMP contributes to angiogenesis. METHODS: MT1 MMP was transfected into human breast carcinoma cell line MCF-7 cells. Semi quantitative RT-PCR and immunofluorescence staining were employed to detect the expression of VEGF in the transfected and non-transfected MCF-7 cells. Tumor growth, microvessel density and expression of VEGF in nude mice were detected through in vivo tumorigenicity assay. RESULTS: In MT1-MMP stable transfected MCF 7 cells, mRNA expression of VEGF(189), VEGF(165), and VEGF(121) and immunofluorescence intensity were significantly elevated (P < 0.001). In vivo tumorigenicity assay in the nude mice showed that MT1-MMP promoted tumor growth. The MVD in the MT1-MMP-transfected cells-transplanted tumor tissue was significantly elevated (P < 0.05). Immunohistochemical assay showed that there was a strong immunostaining of VEGF in those tumor tissues. CONCLUSION: MT1-MMP can induce tumor angiogenesis through up-regulation of VEGF expression. This function of MT1-MMP may open a new approach for clinical anti-tumor research and anti-tumor drug development. PMID- 20021823 TI - [Over-expression of caveolin-1 inhibits proliferation and invasion of pancreatic carcinoma cells in vitro]. AB - OBJECTIVE: To investigate the effects of caveolin-1 on the biologic behavior of pancreatic carcinoma cell line panc1 cells in vitro. METHODS: Eukaryotic expression vectors containing human caveolin-1 gene was stably transfected into panc1 cells with Lipofectamine2000. The clones stably overexpressing caveolin-1 were identified by real-time PCR and Western plotting. The cell growth activity was examined by MTT assay. Anchorage-independent growth was detected by colony formation assay in soft agar. Flow cytometry was used to analyze the cell cycle and apoptosis. Cell invasion assay was used for evaluating cell invasion capacity. The relative phosphorylation level of EGFR, c-Raf, Mek, Erk, p38 and SAPK/JNK were detected by Western blotting. RESULTS: Three transfected cell clones overexpressing caveolin-1 were obtained. Comparing with the panc1 cells, the transfected cells exhibited a slower growth rate and formed fewer colonies in soft agar. The results of flow cytometry showed that over-expression of caveolin 1 resulted in the cell cycle arrest at G(0)/G(1) phase and increased the apoptotic cell fraction. Cell invasion assay showed that overexpression of caveolin-1 significantly inhibited the panc1 cell invasion. Western blotting results showed that overexpression of caveolin-1 reduced the phosphorylation of EGFR, c-Raf, Mek and Erk while did not affect the activity of p38 and SAPK/JNK. CONCLUSION: Over-expression of caveolin-1 inhibits the growth and invasion of pancreatic carcinoma cells in vitro. These phenotypes may be correlated with the inhibition of EGFR-c-Raf-Mek-Erk signaling pathway. PMID- 20021824 TI - [Exosomes derived form bladder transitional cell carcinoma cells induce CTL cytotoxicity in vitro]. AB - OBJECTIVE: To isolate and purify exosomes derived from human bladder transitional cell carcinoma T24 cells, analyze the morphology and protein composition, and investigate the antitumor effect of specific cytotoxic T lymphocytes induced by exosomes. METHODS: Exosomes were isolated and purified by ultrafiltration and sucrose gradient centrifugation, and characterized by electron microscopy and Western blot. Dendritic cells were amplified and purified from peripheral blood and pulsed with exosomes. Then they were co-cultured with T cells, and divided into 3 groups: exosome-pulsed DC group, unplused DC group and control group. Alamar-Blue assay was used to evaluate the specific cytolytic activity. RESULTS: The exosomes were in size about 30 approximately 90 nm saucer-shaped membranous vesicles. HSP70, ICAM-1 and CK20 were detected by Western blot. The CTL induced by DC pulsed with exosomes had significant cytolytic activity (P < 0.01). CONCLUSION: The exosomes derived from T24 cells are loaded with immunoprotein HSP70 and ICAM-1, and DC pulsed with exosomes can promote the anti-tumor effect of CTLs in vitro. PMID- 20021825 TI - [Effect of actovegin on intestinal mucosa and expression of bcl-2/bax genes in rats with acute radiation enteritis]. AB - OBJECTIVE: To evaluate the effect of actovegin (Nycomed, deproteinized hemoderivative of calf blood injection) on intestinal mucosa in rats with acute radiation enteritis, and observe the changes of expression of apoptosis-related bcl-2/bax genes. METHODS: An abdominal irradiation in a dose of 9.0 Gy X-ray of linear accelerator was performed once on a group of Wistar rats to establish a model of acute intestinal radiation enteritis. The experimental rats were randomly divided into five groups. Group 1 was normal control group; group 2 was model control group; groups 3, 4 and 5 were treated with low, middle and high dose of actovegin, respectively. After the model was established, actovegin injection was given intraperitoneally for successive 4 days. Corresponding intestinal tissues were taken for morphological examination with an image analysis system. The expression of apoptosis related bax and bcl-2 protein in the intestinal mucosal epithelial cells was determined by immunohistochemistry. RESULTS: The groups 4 and 5 had significantly higher height of intestinal villi, the depth of crypt, the thickness of the mucosa and entire wall (254.66/261.71 microm, 166.47/165.41 microm, 510.44/511.71 microm, 610.38/608.98 microm), compared with those of the model control group (239.12 microm, 151.45 microm, 420.27 microm and 579.32 microm), respectively (P < 0.05). Treatment with middle and high doses of actovegin also significantly down-regulated the expression of activating apoptosis protein bax (24.54/23.24) compared with that of model control group (59.32) (P < 0.05) and up-regulated the expression of inhibiting apoptosis protein bcl-2 (55.54/52.21) compared with that of model control group (20.32) (P < 0.05). The ratio of bcl-2/bax was significantly higher in the groups 4 and 5 (2.2632, 2.1275) compared with that in the model control group (0.3425) (P < 0.01). CONCLUSION: Actovegin accelerates the recovery of the acute radiation injured intestinal mucosal epithelium by decreasing apoptosis via down-regulation of the expression of activating apoptosis protein bax and up-regulation of inhibiting apoptosis protein bcl-2. PMID- 20021827 TI - [Effects of survivin in tamoxifen-induced inhibition of proliferation and enhancement of apoptosis in hepatoma Hep22 cells]. PMID- 20021826 TI - [Combination with SN-38 on human colon cancer LoVo cells]. AB - OBJECTIVE: To observe the anti-proliferation effect of bevacizumab and SN-38 (active metabolite of irinotecan), and investigate the possible mechanisms of these two agents. METHODS: Human colon cancer LoVo cells were cultured under hypoxic conditions. Inhibition of cell proliferation was evaluated by MTT assay. The drug modulation on HIF-1alpha, VEGF, ERK and AKT were assessed by the following assays. The mRNA expression of HIF-1alpha and VEGF were measured by RT PCR. The protein expression of HIF-1alpha, ERK and AKT were evaluated by Western blot analysis, and VEGF by ELISA assay. RESULTS: Among different combination schedules, Bevacizumab given after SN-38 show most synergistic anti-proliferation effect. Under hypoxic conditions, the expression of HIF-1alpha and VEGF increased as time accumulated, Bevacizumab combined with SN-38 almost completely inhibited the expression of HIF-1alpha and VEGF. Moreover, the MAP kinase pathway was involved in the drug modulation of HIF-1alpha and VEGF. CONCLUSION: These findings suggest the anti-proliferation effect of bevacizumab and SN-38 was schedule-dependent, and the synergistic effect of Bevacizumab and SN-38 was related to drug modulation of the HIF-1alpha and MAP kinase pathway. PMID- 20021828 TI - [Correlation between aneuploidy of chromosome 17, over-expression of TP53 and TOPIIalpha, and the clinicopathological features and diagnosis of gastric adenocarcinoma]. AB - OBJECTIVE: The purpose of this study was to investigate the markers which can be used in auxiliary diagnosis of gastric adenocarcinoma (GAC), and their correlation with their clinicopathological features. METHODS: 122 surgical specimens including 99 gastric adenocarcinoma (GAC), 18 adjacent mucosa and 5 distal normal mucosa were collected, and analyzed by in situ hybridization (FISH). The centromere probe cen17, specific for chromosome 17, which was reported to be frequently amplified in GAC, was selected for the FISH analysis. The clinicopathological features of the 99 GAC cases were reviewed, and the level of TP53 and TOPIIalpha gene expression, located in chromosome 17, was detected using tissue micro-array (TMA), compared with that of corresponding adjacent normal mucosa. Data were analyzed with SPSS 11.5 for Windows. RESULTS: The statistical results of FISH and TMA showed that 58.6% of cen17 in tumor tissues were aneuploid, and 45.5% of TP53 and 84.7% of TOPIIalpha were over-expressed in GAC samples, significantly higher than those in non-tumor gastric mucosa (0, 12.1% and 14.1%, respectively) (P = 0.000). 58 GAC tissues were aneuploid of cen17, including 26 cases TP53-positive and 49 cases TOPIIalpha-positive. The expression of TP53 in non-tumor gastric mucosa with dysplasia was significantly higher than that in the mucosa without dysplasia (P = 0.009). Aneuploidy of cen17 was more frequent in grade 1 or 2 than in grade 3 GAC (P < 0.05). Higher frequency of aneuploidy of cen17 was also observed in the gastric cardia than in pylorus (P < 0.05), while no correlation was found between aneuploidy of cen17 and age, sex of patients, lymph node metastasis, and clinical stage of tumors. Over-expression of TP53 protein was associated with the size of tumors (P < 0.05). In addition, a negative correlation was observed between over-expression of TOPIIalpha and lymph node metastasis (LNM) as well as TNM classification (P < 0.05). CONCLUSION: Detection of aneuploidy of cen17 as well as over-expression of TP53 and TOPIIalpha may be helpful in the diagnosis and prognostic prediction of gastric adenocarcinoma. PMID- 20021829 TI - [Case-control study of risk factors in cholangiocarcinoma]. AB - OBJECTIVE: To investigate the risk factors of intrahepatic cholangiocarcinoma (ICC) and extrahepatic cholangiocarcinoma (ECC). METHODS: The clinicopathological data of 190 patients with cholangiocarcinomas (61 ICC and 129 ECC) diagnosed and treated in the Peking Union Medical College Hospital between 1998 and 2008 were collected. The clinicopathological data of 380 matched healthy controls were also collected. The information about liver diseases, family history, diabetes, smoking and drinking were recorded and analyzed. RESULTS: The positive rate of HBsAg(+) and anti-HBc(+), HBsAg(-) and anti-HBc(+) and the incidence of choledocholithiasis or hepatolithiasis in ICC patients were 27.9%, 50.8% and 14.8%, respectively. The incidence of diabetes mellitus, cholecystolithiasis, choledocholithiasis or hepatolithiasis and previous cholecystectomy in ECC patients were 18.6%, 15.5%, 18.6% and 13.2%, respectively. The incidences of all above mentioned factors in the ICC or ECC patients were significantly higher than that in the controls (P < 0.05). Compared with the patients with ECC, the ICC patients had a significantly higher cirrhosis rate (P < 0.05). CONCLUSION: Our study results show that choledocholithiasis or hepatolithiasis, liver cirrhosis and chronic HBV infection are possible risk factors for intrahepatic cholangiocarcinoma, while choledocholithiasis or hepatolithiasis, diabetes mellitus, cholecystolithiasis, history of cholecystectomy are risk factors for extrahepatic cholangiocarcinoma. PMID- 20021830 TI - [Prognostic significance of metastatic lymph node ratio in colorectal cancer]. AB - OBJECTIVE: To investigate the prognostic significance of metastatic lymph node ratio in patients with colorectal cancer. METHODS: The clinicopathological data of 303 surgically treated patients with colorectal cancer were retrospectively analyzed. Spearman correlation analysis was used to determine the correlation coefficient. The survival was analyzed using Kaplan-Meier method, and the survival difference was assessed by Log-rank test. Multivariate analysis was performed using Cox proportional hazard regression model in forward stepwise regression. Receiver working characteristic curve was used to compare the accuracy of the metastatic lymph nodes ratio in predicting the death of patients at 5 years postoperatively with that of the number of metastatic lymph nodes. RESULTS: The MLR was not correlated with the total number of dissected lymph nodes (Spearman correlation coefficient: -0.099, P > 0.05), but the positive rate of metastatic lymph nodes did (correlation coefficient: 0.107, P < 0.05). Kaplan Meier survival analysis revealed that the MLR significantly influenced the postoperative survival time (Log-rank chi(2) = 42.878, P < 0.01), even in the patients with less than 12 resected lymph nodes. The 5-year survival rates for rN0, rN1, rN2 and rN3 were 90.9%, 68.9%, 54.7% and 39.4%, respectively. There was a significant difference between the different stages (P < 0.01). Cox proportional hazard regression model analysis showed that the metastatic lymph node ratio was an independent prognostic factor. (EXP(B) = 7.809, P < 0.01). There was no significant difference between metastatic lymph node ratio and the number of metastatic lymph nodes in predicting the death of patients at 5 years postoperatively based on the area under the receiver working characteristic curve. CONCLUSION: The metastatic lymph node ratio in colorectal cancer patients is not correlated with the total number of dissected lymph nodes. The metastatic lymph node ratio is a major independent prognostic factor for patients with colorectal cancer. The ability of metastatic lymph node ratio in predicting the death of colorectal cancer patients at 5 years postoperatively is the same as that of the number of metastatic lymph nodes. PMID- 20021831 TI - [Value of thallium-201 scintigraphy in assessment of neoadjuvant chemotherapy for osteosarcoma]. AB - OBJECTIVE: To investigate the significance of Tl-201 scintigraphy for assessment of neoadjuvant chemotherapy for osteosarcoma. METHODS: Thirty-four cases with osteosarcoma were enrolled into this study. The Tl-201 scintigraphy features including uptake ratio(UR) and size ratio(SR) were obtained in both early and delay imaging stages before and after chemotherapy. The responses of chemotherapy were classified into three grades according to the percentage of tumor necrosis in the specimens: necrosis < 50% as grade 1, necrosis of 50% approximately 90% as grade 2, diffuse necrosis > 90% as grade 3. The alteration ratio (AR) and SR were calculated according to tumor necrosis ratio (TNR). RESULTS: Of the 6 patients with grade 1 response, the mean alteration ratios were 3.19% +/- 8.40% and 26.29% +/- 63.61% in early and delay imaging stages, respectively. Among the 18 patients with grade 2 response, the mean alteration ratios were 40.07% +/- 11.95% and 39.30% +/- 9.87%, respectively. Of the 10 patients with grade 3 response, the mean alteration ratios were 78.32% +/- 8.33% and 63.26% +/- 6.06% in early and delay imaging stages, respectively. The results of liner regression analysis of TNR of the surgical specimens showed a significantly positive correlation (r = 0.71) between AR and TNR. The lesion size was reduced in 18 cases, but unchanged in 8 and increased in 8. The liner regression analysis results showed a negative correlation between SR and TNR. CONCLUSION: The alteration ratio changes significantly after neoadjuvant chemotherapy and has a significantly positive correlation with tumor necrosis ratio. Thallium-201 scintigraphy is helpful in the evaluation of neoadjuvant chemotherapy for osteosarcoma. PMID- 20021832 TI - [Correlation between PPARgamma and VEGF-C expression in extrahepatic cholangioadenocarcinoma (EHCAC) and their prognostic significance]. AB - OBJECTIVE: To evaluate the expression of vascular endothelial growth factor C (VEGF-C) and peroxisome proliferators-activated receptors (PPARgamma) in extrahepatic cholangioadenocarcinoma (EHCAC) and to elucidate its correlation with clinicopathological factors and their significance in prognosis. METHODS: The expressions of PPARgamma and VEGF-C were detected by immunohistochemistry in 69 cases of EHCAC, 12 cases of non-tumor bile duct epithelium, and their relationship to clinicopathological parameters and follow-up were analyzed. RESULTS: The positive rate of PPARgamma expression in 69 cases of EHCAC was 59.4%, significantly higher than that in 12 cases of non-tumor bile duct epithelium (0%), (P < 0.01). The positive rate of VEGF-C in 69 cases of EHCAC was 84.1%, also significantly higher than 16.7% in 12 cases of benign bile duct epithelium (P < 0.05). PPARgamma expression was associated with clinical TNM stage and lymph node metastasis. VEGF-C expression was associated with lymph node metastasis. Cox analysis results showed that portal vein and/or hepatic artery invasion, lymph node metastasis and VEGF-C expression were independent prognostic factors of EHCAC (P < 0.05). CONCLUSION: PPARgamma expression may play an important role during tumorigenesis of extrahepatic cholangioadenocarcinoma. The expressions of PPARgamma and VEGF-C are significantly correlated with the clinicopathological characteristics and biological behavior of EHCAC. Expression of VEGF-C is an independent prognosis factors in EHCAC. The detection of PPARgamma and VEGF-C is valuable for evaluation of prognosis of EHCAC. PMID- 20021833 TI - [Association between Ephb4 gene polymorphism and non-small cell lung cancer]. PMID- 20021834 TI - [Evaluation of scalene lymph node or contralateral mediastinum biopsy during mediastinoscopy for non-small cell lung cancer]. AB - OBJECTIVE: To explore the clinical indication of N3 lymph node biopsy during mediastinoscopy for non-small cell lung cancer (NSCLC). METHODS: Cervical mediastinoscopy was performed in 89 patients with clinical stage I-IIIA non-small cell lung cancer prior to thoracotomy. Of those, 12 underwent cervical medistinoscopy combined with right scalene lymph node biopsy and 10 with anterior mediastinotomy. RESULTS: Nine patients were found to have lymph node metastasis (N3 disease) during mediastinosopy. Of those, 6 had contralateral mediastinal lymph node metastasis and 3 cases with right scalene lymph node metastasis. The incidence of N3 disease in the patients with adenocarcinoma, serum CEA > 5 ng/ml and multi-station mediastinal lymph node metastasis was significantly higher than that in those with non-adenocarcinoma, CEA < 5 ng/ml and ipsilateral uni-station mediastinal lymph nodes metastasis (P < 0.05). CONCLUSION: Biopsy of scalene lymph node or contralateral mediastinal lymph node should be performed during mediastinoscopy in order to exclude N3 disease for potentially operable NSCLC patients with adenocarcinoma, serum CEA >5 ng/ml and ipsilateral multi-station mediastinal lymph nodes metastasis. PMID- 20021835 TI - [Comparison of response evaluation methods of neoadjuvant chemotherapy in breast cancer patients]. AB - OBJECTIVE: To compare the efficiency of response evaluation by clinical examination, ultrasonograghy and mammography in neoadjuvant chemotherapy (NAC) for breast cancer. METHODS: A retrospective cohort study was conducted to analyze the data of 141 patients treated with neoadjuvant chemotherapy. Response evaluation was performed by clinical palpation, ultrasound and mammography. RESULTS: Only 12 (8.5%) among the 141 patients presented with a stage I tumor. The tumor size determined by palpation was often larger than that by ultrasound before therapy (P < 0.01). Among patients with suspicions axillary nodes checked by ultrasound, 88.3% (53/60) of them had positive nodes by pathology before NAC, and 34.5% (10/29) of patients with negative nodes determined by ultrasound had positive nodes by pathology. In all the 141 patients, 21(14.9%) showed pathological complete remission in both the primary tumor and lymph node. For response evaluation, the false complete remission rate judged by clinical examination was 46.8% (22/47), and the false tumor residual rate by ultrasound was 84.0% (21/25). In 53.5% (23/43) of patients the response could not be assessed by mammography due to that the tumors were undistinguishable in size. The range of microcalcification was not reduced in 5 patients with a partial response of the tumor. 25 patients experienced needle puncture during therapy. Among them, in the 9 pathologically negative patients, only 3 achieved pCR, and the other 16 positive patients didn't achieve pCR. CONCLUSION: Using the puncture or sentinel lymph node biopsy, clinicians should pay enough emphasis on the pathological determination of the node status before chemotherapy. Clinicians will make a quite of false judgment of the tumor by clinical examination, ultrasound or mammography. They may use needle puncture during therapy to evaluate the response of neoadjuvant chemotherapy, and the result should be analyzed synthetically. PMID- 20021836 TI - [Feasibility study on CT guided percutaneous incisional needle biopsy for deep pelvic masses by different puncture approaches]. AB - OBJECTIVE: To explore the feasibility of CT guided percutaneous incisional needle biopsy (PINB) for deep pelvic masses at different locations via various puncture approaches. METHODS: PINBs under CT guidance were performed in 70 patients with 72 pelvic lesions through different puncture approaches. Their pathological findings and safety were evaluated after follow-up of a period of 1-34 months. RESULTS: PINBs were performed through transpiriform-muscle in 27 cases, 16 through transgluteal approach, 5 through posterior oblique approach in prone position, 8 by anterior or lateral transabdominal route, 8 through iliopsoas muscle and 8 by direct transosseous approach, respectively. Sixty-four malignant lesions were confirmed by pathology, including 30 adenocarcinomas, 19 squamous cell carcinomas, 5 unclassified malignant tumors, 3 small cell carcinomas, 2 malignant giant cell tumors of bone, 2 hepatocellular carcinomas and 3 false negative lesions which were confirmed at the second PINBs as malignant tumors, respectively. Benign neoplasms were confirmed in 8 cases, including fibrosis tissue in 6 lesions, bone tuberculosis in 1 and ovarian cyst in 1. The sensitivity, specificity, and accuracy rate were 95.3% (61/64), 100% (8/8), and 95.8% (69/72), respectively. Twenty-two cases via transpiriform-muscle approach suffered from transient deep pelvic pain which radiated to the lower limbs of the same side. No hematoma, nerve damage, infection, and tumor transplantation in pelvic cavity developed after the PINB procedure. CONCLUSION: CT guided percutaneous incisional needle biopsy through different puncture approaches is safe and feasible for the patients with deep masses at different locations in the pelvic cavity. PMID- 20021837 TI - [Clinical impact of extracapsular extension of axillary lymph node metastases in breast cancer]. AB - OBJECTIVE: To study the clinical significance of extracapsular extension (ECE) of axillary lymph node metastases in breast cancer. METHODS: The clinicopathological data of 1230 cases of nodal positive breast cancer treated in our department from 1989 to 1995 were analyzed retrospectively. RESULTS: 486 (39.5%) from the 1230 cases were ECE positive. There was a higher incidence of ECE in postmenopausal women than premenopausal ones (47.5% versus 35.5%, respectively, P < 0.001). The patients in ECE positive group had a larger tumor size (5.11 +/- 2.53 cm versus 3.90 +/- 1.80 cm, P < 0.001). 18.3% of patients with stage T1 were ECE positive, stage T2 were 36.4%, and stage T3 were 54.4%, and the difference was significant (P < 0.001). ECE was correlated with the number of positive axillary lymph nodes. The ECE positive group had more positive nodes than ECE negative group (16.96 +/- 12.16 versus 5.24 +/- 6.60, P < 0.001). 6.1% of patients with 1 positive node were ECE positive, 13.5% with 2 - 3, 35.8% with 4 - 9, 62.3% with 10 - 19, and 84.0% with more than 20 positive axillary nodes, and there was a significant difference among those groups (P < 0.001). ECE had no association with ER/PR status (P = 0.706). ECE was a risk factor of local-regional recurrence, but the relapse time had no significant difference (P = 0.559). ECE was also a risk factor of distant metastasis, and the relapse time had a significant difference (P < 0.001). The median metastasis free time was 30.0 (2 approximately 172) months in ECE positive group, while 37.5 (2 approximately 170) months in ECE negative group (P = 0.006). CE occurred in 60.4% of the patients with firstly diagnosed bone, skin and distant lymph node metastasis, but in 42.0% of the patients with firstly diagnosed visceral metastasis (P = 0.001). The metastasis free survival rate, locoregional recurrence-free survival rate and overall survival rate of the ECE positive group were much shorter than that of the ECE negative group. COX proportional hazard regression single factor analysis and multi-factor analysis suggested that ECE is an independent factor of metastasis free survival, locoregional free recurrence and overall survival. CONCLUSION: The presence of ECE in breast cancer is positively related with tumor size and the number of positive lymph nodes. It is also a risk factor of locoregional recurrence and distant metastasis. ECE positive group has a much shorter metastasis-free survival, locoregional recurrence-free survival and overall survival. ECE is a risk factor of those three indexes. PMID- 20021838 TI - [Clinical analysis of premedication regimen with dexamethasone at different doses for docetaxel-induced hypersensitivity reaction]. PMID- 20021839 TI - [A look back at the past 30 years of research on precancerous lesions in the stomach in Cancer Institute of China Medical University]. PMID- 20021840 TI - [Particulate matter: an old pollutant with increasing public health challenges]. PMID- 20021841 TI - [Cause-excluded health adjusted life expectancy: a new indicator in measuring the burden of diseases]. AB - Based on the integration of concept and methods of cause-eliminated life expectancy (CELE), health-adjusted life expectancy (HALE) and disability-adjusted life year (DALY), a new index named cause-excluded health adjusted life expectancy (CEHALE) was developed in this study to evaluate the health impact(both fetal and non-fetal) of single disease. In order to include the impact of both premature death and health problems among those who are alive, the concept of suppositional death was introduced to calculate CEHALE. Cause-excluded health adjusted life expectancy developed in this study, should be not only easy to understand in concept, but also has theoretical support and be not affected by age structure, and be more suitable to use in comparison among different diseases and different population. Moreover, the new index is easy to calculate. So, CEHALE should be an ideal index to evaluate the impact of disease to population health. PMID- 20021843 TI - [Pollutions of indoor fine particles in four types of public places and the influencing factors]. AB - OBJECTIVE: To study the levels of pollutions caused by fine particulate matter (PM(2.5)) in the public places and investigate the possible influencing factors. METHODS: A total of 20 public places in four types such as rest room in bath center, restaurant, karaoke bars and cyber cafe in Tongzhou district in Beijing were chosen in this study; indoor and outdoor PM(2.5) was monitored by TSI sidepak AM510. Data under varying conditions were collected and analyzed, such as doors or windows or mechanical ventilation devices being opened, rooms cramped with people and smoking. RESULTS: The average concentration of indoor PM(2.5) in 20 public places was (334.6 +/- 386.3) microg/m(3), ranging from 6 microg/m(3) to 1956 microg/m(3); while in bath center, restaurant, karaoke bars and cyber cafe were (116.9 +/- 100.1)microg/m(3), (317.9 +/- 235.3) microg/m(3), (750.6 +/- 521.6)microg/m(3) and (157.5 +/- 98.5) microg/m(3) respectively. The concentrations of PM(2.5) in restaurant (compared with bath center: Z = -10.785, P < 0.01; compared with karaoke bars: Z = -10.488, P < 0.01; compared with cyber cafe: Z = -7.547, P < 0.01) and karaoke bars (compared with bath center: Z = 16.670, P < 0.01; compared with cyber cafe: Z = -15.682, P < 0.01) were much higher than those in other two places. Single-factor analysis revealed that the average concentration of indoor PM(2.5) in 20 public places was associated with the number of smokers per cube meters(9.13 x 10(-3); r = 0.772, F = 26.579, P < 0.01) and ventilation score [(2.5 +/- 1.5) points; r = 0.667, F = 14.442, P < 0.01], and there were significant correlation between the average indoor and outdoor levels in restaurant [(317.9 +/- 235.3) microg/m(3), (67.8 +/- 78.9) microg/m(3); r = 0.918, F = 16.013, P = 0.028] and cyber cafe [(157.5 +/- 98.5) microg/m(3), (67.7 +/- 43.7) microg/m(3); r = 0.955, F = 30.785, P = 0.012]. Furthermore, significant correlation was observed between the average concentration of indoor PM(2.5) [(157.5 +/- 98.5) microg/m(3)]and the number of people per cube meters (288.7 x 10(-3)) in cyber cafe (r = 0.891, F = 11.615, P = 0.042). Multiple regression analysis showed that smoking (b' = 0.581, t = 3.542, P = 0.003) and ventilation (b' = -0.348, t = -2.122, P = 0.049) were the major factors that may influence the concentration of indoor PM(2.5) in four public places. With cluster analysis, the results showed that the major factors that influence the concentration of indoor PM(2.5) was the outdoor PM(2.5) levels [(49.6 +/- 39.5) microg/m(3); b = 1.556, t = 3.760, P = 0.007] when ventilation (score > 2) was relatively good. The number of smokers per cube meters (14.7 x 10(-3)) became the major influence factor when the ventilation score 0.05). HER2/neu protein level in pSIREN-RetroQ-neu transfected cells was inhibited by 40% and 39% respectively. CONCLUSION: Plasmid-based shRNA expression systems targeted against HER2/neu gene were generated successfully, which resulted in down-regulation of HER2/neu gene expression in the 16HBE-T efficiently. PMID- 20021854 TI - [Efficacy and adverse effects of film coated praziquantel for treatment of schistosomiasis japonica]. AB - OBJECTIVE: To understand the adverse and curative effects of film coated praziquantel for treatment of schistosomiasis japonica. METHODS: A questionnaire survey was conducted among 6 to 65 years old inhabitants from epidemic areas of schistosomiasis japonica in Jiangxi, Anhui and Hubei provinces, while indirect haemagglutination assay (IHA) was also administrated. The Serological positives were tested by Kato-Katz technique. A total of 509 habitats of serological positives and healthy persons were given praziquantel. 460 targets with no relative symptoms were followed up for adverse effect of drugs in one month and efficacy evaluation of praziquantel was carried out in 104 parasitological positives 3 months post-treatment. RESULTS: The 84.7% (144/170) individuals thought film coated tablets were no smell or only with little smell while 92.9%(315/339) targets in control group fell that tablets smelled bad. The total side effect rates of film coated praziquantel group and control group in 1-2 hours post-treatment were 20.30% (27/133) and 83.18% (272/327) respectively. There was significantly different between two groups (chi(2) = 164.316, P < 0.05). The adverse effect rates of film coated praziquantel group in neuromuscular system, digestive system and cardiovascular system were 15.79% (21/133), 9.77% (13/133), 2.26% (3/133) individually which were significantly lower than those of control groups with 81.65% (267/327), 49.24% (161/327), 12.84% (42/327) in corresponding systems (chi(2)(neuromuscular system) = 175.188, chi(2)(digestive system) = 62.601, chi(2)(cardiovascular system) = 12.010, P < 0.05). And the adverse effect rate of allergic reaction of film coated group was no significant difference with control group (2.26%(3/133) versus 0.92% (3/327), chi(2) = 1.315, P = 0.235). One day after treatment, the adverse effect rate of film coated tablets of praziquantel decreased to 3.01% (4/133), significantly lower than that of control group (P < 0.05). There was no significant difference after 2 weeks treatment while the adverse effect rates decreased to 0.75%(1/133), 0.61% (2/327) in film coated praziquantel group and control group respectively (chi(2) = 0.029, P = 0.642). The curative rates 3 months post treatment were 84.91% (45/51) in film coated tablets of praziquantel group and 82.35% (42/53) in control group without significant difference (chi(2) = 1.536, P = 0.215). CONCLUSION: The smell and adverse effects of film coated tablets of praziquantel were decreased significantly and its efficacy for treatment of schistosomiasis japonica was equal to the control tablets. The film coated tablets of praziquantel might be applied in field widely after a further verification. PMID- 20021855 TI - [Comparative analysis of measles genome between vaccine strain and wild-type strain in Zhejiang province of China]. AB - OBJECTIVE: To explore the distinction between wild-type strain MVi/Zhejiang, CHN/7.05/4 and vaccine strain Shanghai-191 at genome level. METHODS: After sequencing of measles wild-stain MVi/Zhejiang. CHN/7.05/4, the distinction between the wild-type strain and the vaccine strain was analysed by MEGA 3.1 software at genome level, and the antigen variation was studied by means of combining the epidemiological data. RESULTS: There were 822 nucleotide differences (5.17%) and 161 amino acid differences between these two strains, including three glycosylation sites variation found. Meanwhile, the antigen ratio between wild-type strain and vaccine strain was found to be 5.66. CONCLUSION: There should be certain differences between the contemporary wild-type strain MVi/Zhejiang, CHN/7.05/4 and vaccine strain Shanghai-191 at genome level, and the protective effects of measles vaccine should be studied further. PMID- 20021856 TI - [Bemusement and strategy on the efficacy of clinical application of targeted anticancer drugs]. PMID- 20021857 TI - [Epigallocatechin gallate induces apoptosis in human hepatocellular carcinoma HepG2 cells via TGF/Smad signaling pathway]. AB - OBJECTIVE: To investigate the cytotoxic effect of epigallocatechin gallate (EGCG) on human hepatocellular carcinoma cell line HepG2 cells and corresponding changes of TGF-beta1-Smad pathway. METHODS: The cytotoxic effect of EGCG on HepG2 cells was determined by MTT assay. Cell cycle and apoptosis rate were detected by flow cytometry. RT-PCR and luciferase assay were used to verify whether TGF-beta1-Smad signaling pathway is intact in HepG2. The mRNA expression of Smad 2, Smad3, Smad4 and Smad7 was detected by real-time PCR. RESULTS: EGCG induced apoptosis in the HepG2 cells in a time- and concentration-dependent manner. The proportion of G(1) phase cells was increased gradually as the concentration increased. However, the percentage of cells in S phase was decreased gradually. Annexin V/PI assay demonstrated that early apoptosis increased as the concentration increased, and late apoptosis also increased, when treated with high-concentration EGCG. The intact TGF-beta1-Smad pathway was verified by luciferase assay and RT-PCR. There was no significant effect of EGCG on mRNA level of Smad 2, Smad 3, and Smad 4 in HepG2 cells, but downregulated mRNA level of Smad 7. CONCLUSION: EGCG can reduce apoptosis in human hepatocellular carcinoma cell line HepG2 cells. The activation of TGF-beta1-Smad signaling pathway may be involved in its cytotoxicity mechanisms. PMID- 20021858 TI - [Apoptosis resistance induced by leptin and its mechanism in breast cancer cells]. AB - OBJECTIVE: To explore the apoptosis resistance induced by Leptin and its mechanism in breast cancer cells in vitro. METHODS: The leptin-mediated reduction of docetaxel-induced apoptosis in human breast cancer T47D cells was evaluated by TransAM ELISA, MTT and caspase-9 assay. The leptin-promoted survivin expression was analyzed by Western-blot and RT-PCR. The reversing effect of STAT3 knockdown on leptin-induced survivin upregulation was measured by Western-blot and RT-PCR. RESULTS: Leptin promoted T47D cells proliferation and the inhibitory rate was 63.6%. It reduced docetaxel-induced apoptosis in T47D cells by 31.9%. Leptin at different concentrations promoted survivin protein and mRNA expression in T47D cells. The expression of survivin mRNA was 4.6 fold compared with the T47D cells not treated with leptin(10 nmol/L). The expression of survivin mRNA in T47D cells was 0.55 +/- 0.15 fold after transfected with small interfering RNA (siRNA) of STAT3. The expression of survivin mRNA in STAT3 siRNA group and mock transfected group were 0.56 +/- 0.18 fold and 1.61 +/- 0.22 fold after treated by leptin, respectively. The survivin protein level of T47D mock transfected cells was increased after treated by leptin, but the protein level of T47D transfected with STAT3 siRNA cells were not changed significantly. CONCLUSION: Leptin/STAT3 signaling is a novel pathway for up-regulation of survivin expression in breast cancer cells. PMID- 20021859 TI - [Preliminary study of proteins related to blast crisis in chronic myeloid leukemia]. AB - OBJECTIVE: To identify and compare the expression profiles of differential proteins between chronic phase and blast crisis in chronic myeloid leukemia (CML) by proteomic analysis, and screen the proteins related to blast crisis. METHODS: The total cellular proteins from the bone marrow cells at chronic phase (CP) and blast crisis (BC) in CML were separated by two-dimensional polyacrylamide gel electrophoresis (2-DE) and analyzed with ImageMaster 5.0 software to screen the differential protein spots. Differential protein spots were identified by mass spectrometry for peptide mass fingerprint in combination with database searching from SWISS-PROT. Then 3 protein spots were selected to verify at protein and mRNA levels by Western blot and semi-quantitative RT-PCR, separately. RESULTS: Comparing gel pages from CML-CP and CML-BC, the expression of 13 protein spots decreased and 25 protein spots increased significantly in CML-BC. Twenty differential protein spots were identified by mass spectrometry and 15 were successfully determined. The results of Western blotting were similar to those of 2-DE and showed a high expression of hnRNPK, annexin A1 and RhoA. Semi quantitative RT-PCR analysis showed that there was no correlation between the protein expression changes and mRNA levels of hnRNPK, annexin A1 and RhoA. CONCLUSION: A group of proteins associated with blast crisis are obtained and the results may provide clues for further research to elucidate the role of these proteins in CML-BC carcinogenesis and to develop potential associated biomarkers. PMID- 20021860 TI - [Relationship between the level of RRM1 expression and the sensitivity to gemcitabine in the esophageal squamous cell carcinoma cell lines]. AB - OBJECTIVE: Ribonucleotide reductase subunit M1 (RRM1) is the intracellular target of gemcitabine (GEM). The aim of this study is to explore the relationship between the level of RRM1 expression and the sensitivity to GEM in the esophageal squamous cell carcinoma cell lines. METHODS: Four esophageal squamous cell carcinoma cell lines (Kyse-150, Kyse-450, 9706 and Eca-109) were cultured in vitro. In the same period, RRM1 expression level was measured by RT-PCR and Western blot, and cell sensitivity to GEM was determined by CCK-8 assay. The relation between cell sensitivity and RRM1 expression was further analyzed. Kyse 450 cells were continuously cultured in the medium containing 50 nM GEM. RRM1 expression was measured at different time points to monitor the dynamic changes in the surviving cells. Inhibition of RRM1 expression by RNAi method was applied and the effect on GEM-sensitivity was further examined. RESULTS: The IC(50) of Eca-109, Kyse-150, Kyse-450 and 9706 cells were (0.92 +/- 0.17), (0.48 +/- 0.11), (0.29 +/- 0.06) and (0.02 +/- 0.01) mmol/L, respectively. The expressions of RRM1 protein and mRNA of Eca-109 cell line were the highest detected by Western blot and RT-PCR, followed by Kyse-150 and Kyse-450, and the lowest one was 9706 cell line. When Kyse-450 cells were continuously treated with 50 nmol/L GEM, the level of RRM1 protein was increasing in the surviving cells. RRM1 siRNA could effectively knock down the expression of RRM1 and significantly increase the cell sensitivity to GEM (P = 0.035). CONCLUSION: The level of RRM1 expression correlates with the cell sensitivity to gemcitabine. The cells with a lower level of RRM1 expression are more sensitive to gemcitabine. PMID- 20021861 TI - [Estimation of cancer incidence and mortality in China in 2004-2005]. AB - OBJECTIVE: To estimate the cancer incidence and mortality in China in 2004 - 2005 based on available resources. METHODS: The cancer mortality and incidence ratios were obtained on the basis of national cancer registration database in 2003 and 2004 adjusted by age and area. The crude cancer mortalities were retrieved from the database of the third national death survey 2004-2005. Incidence rates of specified cancer types were calculated using mortality and M/I ratios. RESULTS: The total estimated cancer incidence and mortality per year in 2004 - 2005 were 2 596 112 new cases and 1 798 147 deaths, both were higher in males than in females. There were 1 337 227 new cases and 841 860 deaths in urban areas compared with 1 258 885 new cases and 956 287 deaths in rural areas. Cancers of the lung, stomach, liver, esophagus and colorectum/anus were the commonest cancers in China with 483 040, 428 380, 370 236, 236 589 and 197 873 new cases, respectively. Those five cancers were leading causes of cancer death with a number of 420 411, 339 308, 318 756, 190 233 and 101 684, respectively. CONCLUSION: Cancer burden in China is getting more and more serious. It is imperative to enhance effective cancer prevention and control in China. PMID- 20021862 TI - [Correlations of 99 Tc m-HL91 SPECT hypoxia imaging with HIF-1alpha and VEGF expression in non-small cell lung cancer]. AB - OBJECTIVE: (99)Tc(m)-HL91 ((99)Tc(m) labeled 4, 9-diaza-3, 3, 10, 10 tetramethyldodecan-2, 11-dione dioxime) is a potential noninvasive marker of tumor hypoxia. It has been reported that (99)Tc(m)-HL91 has validity for hypoxia imaging in non-small cell lung cancer (NSCLC). The aim of this study was to evaluate the (99)Tc(m)-HL91 SPECT hypoxia imaging of NSCLC, the expression of inducible hypoxia factor-1alpha (HIF-1alpha) and vascular endothelial growth factor (VEGF), and to analyze their correlations with clinicopathological characteristics. METHODS: Twenty NSCLC patients who underwent radical resection were enrolled into this study prospectively. (99)Tc(m)-HL91 SPECT scanning was performed in all patients at one or two days before surgery. After intravenous injection of approximately 740 MBq (99)Tc(m)-HL91, anterior, posterior and lateral planar images were collected at 2, 4 and 6 hours, respectively. Regions of interest (ROIs) were drawn in the tumor and the contralateral normal lung tissue, and the radioactivity ratio of tumor to normal tissue (T/N) was calculated. Immunohistochemistry was used to detect the expression of HIF-1alpha and VEGF in sequential histological sections of specimens. RESULTS: Among the 20 NSCLC patients, 13 showed positive expression of HIF-1alpha and 15 had positive expression of VEGF, with a positive rate of 65.0% and 75.0%, respectively. The uptake of (99)Tc(m)-HL91 was strongly correlated with the expression status of HIF-1alpha. No correlation between HIF-1alpha and VEGF expression levels was observed. The HIF-1alpha expression level was not correlated with histological subtype, but with lymph node involvement. The expression levels of HIF-1alpha and VEGF were positively correlated with tumor stage. CONCLUSION: The result of (99)Tc(m)-HL91 SPECT hypoxia imaging is found to be positively correlated with expression of HIF-1alpha in the non-small cell lung cancer. HIF-1alpha expression is positively correlated with VEGF expression. Furthermore, both HIF-1alpha and VEGF expressions are increasing with the increase of tumor stage. PMID- 20021863 TI - [Relationship between Ets-1 expression and angiogenesis, clinicopathological features and survival of patients with gastric carcinoma]. AB - OBJECTIVE: To investigate the expression of Ets-1 in gastric carcinoma, para cancerous tissue and metastatic lymph nodes, and to determine the relationship between Ets-1 expression and clinicopathological features, angiogenesis and survival of patients with gastric carcinoma. METHODS: Gastric carcinoma tissue microarray was used to determine Ets-1 protein expression by SP immunohistochemical staining in 189 advanced gastric cancer, 54 papacancerous tissues, 41 metastatic lymph nodes and 32 control tissues. RESULTS: The positive rates for Ets-1 expression of the carcinoma, paracancerous and control tissues were 71.4%, 29.6% and 18.8%, respectively, with a significant difference among the three groups (P < 0.01). In the cancer tissues, the positive rate of Ets-1 protein expression was significantly associated with depth of invasion and lymph node metastasis (P < 0.01), but not associated with degree of differentiation, Lauren's histological type, sex, age, and size of tumor (P > 0.05). The positive rates for Ets-1 expression of the 41 gastric cancer and 41 metastatic lymph nodes were significantly different (P < 0.05). In metastatic lymph nodes, the positive rate for Ets-1 expression was higher. The MVD in Ets-1 positive tumors was higher than that in the Ets-1 negative tumors, with a significant difference (P < 0.05). Kaplan-Meier survival analysis showed that the survival time of Ets-1-negative patients was longer than that of Ets-1-positive patients (P < 0.05). Cox regression analysis showed that Ets-1 expression was not an independent prognostic factor of gastric carcinoma. CONCLUSION: A higher expression of Ets-1 is involved in carcinogenesis, development, invasion, and metastasis of gastric cancer. Ets-1 plays an important role in angiogenesis in gastric cancer. Ets-1 is a useful marker for predicting the outcome for patients with gastric carcinoma, though it is not an independent prognostic indicator. PMID- 20021864 TI - [Expression of TGF-beta1 and the mechanism of invasiveness and metastasis induced by TGF-beta1 in breast cancer]. AB - OBJECTIVE: To investigate the expression of MMP-2, TIMP-2, TGF-beta1 and TGF betaRI and the relationship among them in breast cancer. METHODS: The protein expression of MMP-2, TIMP-2, TGF-beta1 and TGF-betaRI was detected on tissue chips by S-P immunohistochemical staining in 160 cases of breast carcinoma. RESULTS: The positive rates of TGF-beta1, TGF-beta1 mRNA, MMP-2, MMP-9, TIMP-1 and TIMP-2 expression were 73.7%, 56.2%, 96.9%, 95.0%, 87.5% and 89.4%, respectively. Axillary lymph node metastasis and TNM staging (P < 0.01 and P < 0.01, respectively) were positively correlated to the expression of TGF-beta1. Relase-free survival of TGF-beta1 positive group was lower than that of TGF-beta1 negative group (P = 0.023). The expression of MMP-2 or MMP-9 was positively correlated to that of TGF-beta1 (r = 0.170, P < 0.05; r = 0.221, P < 0.01) and was negatively correlated to that of TGF-beta1 mRNA (r = -0.126, P > 0.05;r = 0.019, P > 0.05). CONCLUSION: The expression of TGF-beta1 may be closely correlated with the invasion and metastasis of breast cancer. TGF-beta1-induced invasiveness and metastasis of breast cancer cells are mediated by MMP-2 and MMP 9. PMID- 20021865 TI - [Characterization of hormone receptor status in 5758 Chinese females with breast cancer]. AB - OBJECTIVE: To analyze the characteristics of hormone receptor status in Chinese females with breast cancer. METHODS: The clinicopathological data of 5758 female breast cancer patients surgically treated in our breast cancer center from Jan. 1997 to Oct. 2008 were retrospectively analyzed. RESULTS: The positive rates of estrogen receptor (ER) and progesterone receptor (PR) were 64.1% and 70.2%, respectively. The ER positive rate was significantly higher in elderly, post menopausal females with a smaller tumor and well-differentiated histology (P < 0.05), while the PR positive rate was significantly correlated with only histological differentiation and tumor size (P < 0.05). The ER and PR positive rates were significantly higher in the patients with lymph node metastasis than that in those without (P < 0.05). Multivariate analysis showed that the histological differentiation, T stage, N stage and menopause status were significantly correlated with ER positive rate, while histological differentiation, T stage and N stage were significantly correlated with PR positive rate. CONCLUSION: Our results show that the ER positive rate of breast cancer in Chinese women is lower than that in western high incidence areas. The ER positive rate is significantly correlated with age, histological differentiation, tumor size, and menopause status. The PR positive rate is correlated only with histological differentiation and tumor size. Interestingly, the ER and PR positive rates are significantly higher in the patients with axillary lymph node metastases than that in those without. However, further study is needed to verify this special phenomenon. PMID- 20021866 TI - [Expression of COX-2 and VEGF and their correlation with angiogenesis in human clear cell renal cell carcinoma]. AB - OBJECTIVE: To investigate the expression of COX-2 and VEGF in clear cell renal cell carcinoma (CCRCC) and their correlation with tumor angiogenesis. METHODS: Envision immunohistochemistry was used to determine the expression of COX-2 and VEGF, and microvessel density (MVD) was marked by CD34 in 80 CCRCC tissues and 20 normal kidney tissues. The relationship between the above mentioned markers were analyzed. RESULTS: Expressions of COX-2 and VEGF were noted in both CCRCC and normal kidney tissues. The positive rates of COX-2 and VEGF were significantly higher in CCRCC than in normal kidney (P < 0.05); The expression of COX-2 was correlated with TNM stage (P < 0.05), histological grade (P < 0.05) and lymph node metastasis (P < 0.05) in CCRCC, but not with age (P = 0.663) and diameter of tumor (P = 0.528). Both COX-2 expression (r = 0.851, P < 0.01) and VEGF expression (r = 0.736, P < 0.01) were significantly associated with MVD in CCRCC. There was a positive correlation between expression of cox-2 and that of VEGF in CCRCC. CONCLUSION: COX-2 expression is correlated with tumor angiogenesis in CCRCC. It is likely that VEGF is one of the most important mediators in the COX-2 angiogenic pathway. PMID- 20021867 TI - [Diagnostic value of multislice spiral CT and MRI in detection of tumor recurrence after liver transplantation for hepatocellular carcinoma]. AB - OBJECTIVE: To investigate the manifestation and diagnostic value of multislice spiral CT (MSCT) and MRI imaging in detection of tumor recurrence after liver transplantation for hepatocellular carcinoma (HCC). METHODS: The clinical data of 161 consecutive HCC patients who underwent orthotopic liver transplantation were retrospectively reviewed. Twenty-nine HCC patients were classified by pTNM according to the "Pittsburgh criteria". MSCT and MRI findings of tumor recurrence after liver transplantation were evaluated retrospectively in 29 stage II-IVb HCC patients. The recurrence site and relapse interval between liver transplantation and recurrence were analyzed. RESULTS: Lung tumor recurrence were found in 21 cases, presented as cotton-like lesions in a diameter of 2 - 3 cm, with a clear margin and homogeneous density. Pleural tumor recurrence was detected in 4 cases. Liver tumor recurrence were found in 9 cases, which can be divided into four subtypes: multinodular in 4 cases, diffuse lesion in 2 cases, huge mass in 2 cases, and uninodular in 1 case. Two cases showed tumor thrombus in the inferior vena cava and portal vein. Lymph node tumor recurrence was found in 9 cases, presented as multiple nodules at hepatic hilum, lesser peritoneal sac, posterior mediastinum, retroperitoneum, or around pancreatic head, and accompanied with merging and necrosis in one case. Bone tumor recurrence were found as osteolytic destruction in 4 cases, and accompanied with adjacent soft-tissue mass in 2 cases. The recurrence sites of the 29 cases were as following: lung (21 cases, 72.4%), liver (9 cases, 31.0%), lymph nodes (9 cases, 31.0%), bone (4 cases, 13.8%) and other sites (3 cases, 10.3%). Lung tumor recurrence was found in all the 10 stage IVb patients with tumor recurrence after liver transplantation, significantly more frequent than that in stage IVa patients (P = 0.023). After liver transplantation, all 25 patients with stage III approximately IVb HCC developed recurrence within one year, but in the 4 cases with stage II HCC at one year later (P = 0.009). CONCLUSION: The results of our study show that in hepatocellular carcinoma patients after liver transplantation, the lung and pleura are the most frequent site of recurrence, followed by liver, lymph node and bone as the second and third sites. The Stage IVb hepatocellular carcinoma should be regarded as a contradiction for liver transplantation due to rapid recurrence. Tumor recurrence occurs later in stage II HCC than in stage III approximately IVb patients. MSCT and MRI are of significant importance in diagnosis and formulating operation plan in HCC patients with recurrence after liver transplantation. PMID- 20021868 TI - [CT and MRI imaging features of peripheral primitive neuroectodermal tumors]. AB - OBJECTIVE: To investigate the CT and MRI features of peripheral primitive neuroectodermal tumors (pPNETs) and evaluate its diagnostic value. METHODS: The clinicopathological data of 9 surgically treated patients with peripheral primitive neuroectodermal tumors confirmed by pathology were collected, spiral CT (4/9) and MRI (6/9) plain scanning and dynamic enhancement scanning were performed preoperatively. Both CT and MRI scannings were performed in 1 case. Those CT and MR images were retrospectively reviewed and analyzed together with clinicopathological findings. RESULTS: The 9 lesions were located in skeletal muscles (n = 6), pelvic cavity (n = 2) and thoracic cavity (n = 1). The tumor size was 7.4-18.3 cm in diameter with a mean diameter of 11.6 cm. The shape of those lesions was round or ellipse (4 lesions) and irregular (5 lesions). The tumor usually presented as ill-defined masses, with homogeneous (n = 2) or inhomogeneous density (n = 7). Seven cases, including the 3 lesions located in the chest and pelvis, showed obvious necrosis and multilocular cyst formation. The tumors showed iso-density as that of the adjacent muscles on CT plain scans and moderate heterogeneous enhancement after intravenous injection of contrast agents. The features of the tumors on the MRI including slightly low signal intensity on SE T1-weighted imaging, iso-signal intensity or slightly high signal intensity on FSE T2-weighted imaging and heterogeneous dynamic delayed contrast enhancement with obvious necrosis in most of them. Six cases had a lesion in the skeletal muscles, presented as a giant ill-defined masse surrounding bone and extended along neural route with bone destruction to varying degrees. CONCLUSION: Primitive neuroectodermal tumor is a kind of malignant tumor with proliferation of small, undifferentiated neuroectodermal cells, usually occurring in children or adolescent and frequently located in the extremities, chest cavity, pelvic cavity and chest wall. It typically presents as a large, ill-defined masse extending along neural route with heterogeneous and obvious enhancement after intravenous injection of contrast agents. The tumors located in the chest and pelvic cavities and some in the extremities show obvious necrosis and multilocular cyst formation, while some of the tumors in the extremities appear as homogeneous solid masses. PMID- 20021869 TI - [Diagnostic value of double contrast-enhanced ultrasonography in preoperative staging of gastric cancer]. AB - OBJECTIVE: To evaluate the clinical value of double contrast-enhanced ultrasonography using oral and intravenous contrast agents in preoperative staging of gastric cancer. METHODS: Sixty-two patients with biopsy-proven gastric cancer were enrolled into this study, and were examined by double contrast enhanced gastric ultrasonography preoperatively. The results were compared with postoperative pathologic findings. RESULTS: The accuracy of oral contrast enhanced gastric ultrasonography and double contrast-enhanced ultrasonography in determining the T stage of gastric cancer was 72.9% (T1: 66.7%, T2: 60.0%, T3: 76.9%, T4: 71.4%) and 88.1% (T1: 66.7%, T2: 80.0%, T3: 89.7%, T4: 100%), respectively, with a statistically significant difference between the two methods (P = 0.036). The sensitivity, specificity, accuracy and Youden index of oral contrast-enhanced gastric ultrasonography and double contrast-enhanced ultrasonography in assessment of lymph node metastasis were 74.5%, 66.7%, 72.9%, and 0.41 versus 89.4%, 75.0%, 86.4%, 0.76, respectively. No significant difference in the accuracy of assessment for lymph node metastasis was observed (P > 0.05). CONCLUSION: Double contrast-enhanced ultrasonography is useful for preoperative staging of gastric cancer, especially for T staging. PMID- 20021870 TI - [Clinical value of prostate specific antigen screening in early detection of prostate cancer]. AB - OBJECTIVE: To evaluate the clinical significance of prostate-specific antigen (PSA) screening in early detection of prostate cancer in Chinese men. METHODS: PSA screening was performed in 8562 asymptomatic men who had been enrolled for health checkup and all were > or = 50 years old. Prostate biopsy was recommended for those with a serum PSA level > or = 4.0 ng/ml. The pathological and clinical features of the patients with prostate cancer detected by the PSA screening were compared with that of 82 clinically diagnosed prostate cancer patients during the same period. RESULTS: Of the 8562 asymptomatic men, 719 had PSA levels > or = 4.0 ng/ml and biopsy was performed in 295 of them. Fifty-eight prostate cancers were detected. The biopsy rate was 41.0% and positive detection rate was 19.7%. The overall age distribution in the screening group and the clinical groups was not significantly different (P = 0.176). However, 41.4% (24/58) of the patients in screening group were > 75 years old, and significantly more than that in the clinical group (25.6%, P = 0.0491). The proportion of the patients with PSA levels > or = 20 ng/ml in the screening group was significantly less than that in the patients of the clinical group (44.8% vs. 75.6%, P = 0.0002). Whether in the patients whose age was > 75 years old (P < 0.05) or < or = 75 years old (P = 0.0002), the patients in the screening group had significantly lower Gleason scores < 7 (60.3% vs. 34.1%, P = 0.002), more T1 or T2 tumor (87.9% vs. 26.8%, P < 0.0001) and more chance to receive radical prostatectomy (50.0% vs. 18.3%, P < 0.0001) than the patients in the clinical group did. However, the distributions of PSA levels at diagnosis and biopsy Gleason scores were not significantly different between the above mentioned two groups (P > 0.05). CONCLUSION: Prostate specific antigen (PSA) screening is useful for early detection of prostate cancer in Chinese men aged > or = 50 years. The patients detected by PSA screening usually show a lower PSA level, Gleason scores and early clinical stage disease, and have more chance for radical prostatectomy than the clinically diagnosed patients. PMID- 20021871 TI - [Analysis of treatment and prognosis of recurrent epithelial ovarian carcinoma]. AB - OBJECTIVE: To discuss the prognostic factors of recurrent ovarian epithelial carcinoma and to analyze the curative effect of post-relapse treatment. METHODS: The clinical records of 293 patients with ovarian epithelial carcinoma were reviewed retrospectively. There were 199 recurrent cases during the following up. RESULTS: All the 199 patients received chemotherapy. And 173 patients only received chemotherapy. 16 patients received surgery and chemotherapy and the other 10 patients received radiotherapy and chemotherapy. 158 patients received platinum-based chemotherapy again and 41 patients received chemotherapy without platinum. The response rate of all the patients was 43.7% (87/199), the response rate of only chemotherapy was 39.9% (69/173), the response rate of surgery and chemotherapy was 75.0% (12/16), and the response rate of radiotherapy and chemotherapy was 60.0% (6/10). The patients were divided into four groups according to the progression free interval (PFI). The response rates in groups that PFI < or = 6 months, 7 - 12 months, 13 - 24 months and > 24 months were 5.1%, 47.2%, 82.1% and 96.0%, respectively. The median survival time in the 16 patients received second cyto-reductive surgery was 41 months. Multivariate analysis revealed that PFI was significantly correlated with prognosis of recurrent ovarian epithelial carcinoma (OR = 0.589, P = 0.021). CONCLUSION: PFI is an individual prognostic factor for survival of recurrent ovarian epithelial carcinoma. PFI is significantly associated with the response rate of chemotherapy. Optimal secondary cytoreductive surgery may improve the overall survival of recurrent patients. The response rate of paclitaxel plus platinum chemotherapy in platinum-sensitive patients is higher than that of other platinum based chemotherapy. PMID- 20021872 TI - [Clinical observation of 21 cases of metastatic renal cell carcinoma treated with sorafenib]. PMID- 20021873 TI - [Clinical observation of the treatment with combination of transcatheter arterial chemoembolization and sorafenib for hepatocellular carcinoma with lung metastasis]. AB - OBJECTIVE: To evaluate the safety and efficacy of the combination of transcatheter arterial chemoembolization (TACE) and sorafenib in treatment of hepatocellular carcinoma (HCC) with lung metastasis. METHODS: Thirty HCC patients with lung metastasis were treated by the combination of TACE and sorafenib between Oct 2006 and May 2008, including 27 men and 3 women. The age of the patients ranged 32 to 73 years old. Sorafenib was administrated orally at 400 mg, twice daily (the less tolerant patients received 200 mg, bid.), if there was no counterindication, at 3 - 4 weeks after TACE, with every 4 weeks as a course of treatment. The efficacy was evaluated at the end of every course of treatment. RESULTS: The metastatic lesions in the lung were diminished in 6 cases and stable diseases achieved in 8 cases. The primary liver tumors were stable in 22 cases, including 10 cases achieved by TACE before sorafenib treatment. Eight cases had slightly progressed liver tumors and were treated with 1 - 3 times of TACE in combination with sorafenib. Side effects included skin lesions in 7 cases, hair loss in 6 cases, fatigue in 18 cases, diarrhea in 6 cases, anemia and bone marrow suppression in 5 cases, high blood pressure in 2 cases, and gastrointestinal bleeding in 1 case. CONCLUSION: The combination of TACE and sorafenib can be used as an effective treatment for hepatocellular carcinoma patients with lung metastasis, which may stabilize the disease in some patients. PMID- 20021874 TI - [Some suggestions on the realization of cancer genome-wide association studies in China]. PMID- 20021875 TI - [Several notable problems in current development of cataract surgery]. AB - The technologies about cataract surgery and intraocular lens has been developing expeditiously. However, some problems have been occurring and should be corrected. Cataract blindness can not restore the sight after surgery has been performed still existed. Some new technologies blindly introduced and applied. The comprehension about individualized selection of intraocular lens still has some bias. To solve the above problems and to hold rhythm of popularizing new techniques and to stick the key points of the technologies may healthily promote and advance the cataract surgery in China. PMID- 20021876 TI - [To pay attention to clinical limitations of accommodative and multifocal intraocular lens]. AB - Clinical application of accommodative and multifocal intraocular lens (IOL) were comparatively successful in recent years. However, we must notice the limitations of these kinds of IOL and the related clinical problems, such as very limited accommodative ability of accommodative IOL and glare or haloes and reduced contrast sensitivity of multifocal IOL. We must pay attention to these clinical problems from these IOL and the unexpected prognosis of these kinds of lens. We must choose the indications strictly, and avoid the misuse. PMID- 20021877 TI - [Quality of vision in eyes that underwent implantation of ReSTOR apodized diffractive multifocal intraocular lens on bilateral eyes in cataract surgery]. AB - OBJECTIVE: To compare the visual outcomes and subjective parameters in patients who underwent implantation of ReSTOR apodized diffractive multifocal intraocular lens on bilateral eyes with those in patients who had monofocal intraocular lens in cataract surgery. METHODS: Retrospective observational case series. 50 eyes of 25 patients received ReSTOR multifocal intraocular lens (multifocal group) and 56 eyes of 28 patients taken implantation of Natural monofocal intraocular lens (monofocal group) participated in the study. The distance and near visual acuity were compared, as well as contrast and glare sensitivity. Visual symptom and spectacle dependence were assessed using a standardized questionnaire. The minimal follow-up was 3 months. RESULTS: An uncorrected distance visual acuity of 0.6 or better was achieved in 94% of eyes in multifocal group and 96% in the monofocal group, respectively (chi(2) = 0.347, P > 0.05). Uncorrected near visual acuity was J(3) or better in 88% of eyes in the multifocal group and 13% in the monofocal group (chi(2) = 60.315, P < 0.01). Best distant corrected near visual acuity was Jr3 or better in 90% of eyes in the multifocal group and 11% in the monofocal group (chi(2) = 66.515, P < 0.01). No significant difference in contrast and glare sensitivity was found between the groups (P > 0.05). Glare was reported as severe in 12% of the multifocal group and 7% of the monofocal group, respectively (chi(2) = 0.365, P > 0.05). Halo was reported as severe in 8% of the multifocal group and 4% of the monofocal group, respectively (chi(2) = 0.485, P > 0.05). 96% of patients in both groups never had to wear glasses for the distant purpose (chi(2) = 0.007, P > 0.05), and 80% in multifocal group and 11% in monofocal group never had to wear glasses for the near purpose (chi(2) = 25.811, P < 0.01). CONCLUSION: The ReSTOR apodized diffractive intraocular lens provided predictably excellent uncorrected distance and near visual acuities, and obtained satisfactory quality of vision. Spectacle independence was significantly higher with this multifocal intraocular lens. PMID- 20021878 TI - [Treatment of postoperative endophthalmitis following cataract surgery without intraocular lens removal]. AB - OBJECTIVE: To explore the effects of vitrectomy combined with silicone oil injection on the treatment of postoperative endophthalmitis following cataract surgery without intraocular lens removal, and to analyze the relative factors. METHODS: This was a retrospective case series study. Clinical data of 7 eyes of 7 patients with postoperative endophthalmitis following cataract surgery underwent the treatment of vitrectomy combined with silicone oil injection without intraocular lens removal from 2003 to 2008 were collected. The outcomes of vision, slit lamp examination, direct and indirect ophthalmoscopy, IOP, and B scan were observed, and the surgical effects were analyzed. RESULTS: Five patients were male, and 2 patients were female. The age ranged from 67.0 to 84.0 years with a mean of 70.0 + or - 4.5. The onset of endophthalmitis ranged from 1 to 3 days with a mean of 2 days. Silicone oil was removed in 5 eyes 3 to 6 months postoperatively. Preoperative visual acuity ranged from non light perception to hand moving. The mean preoperative intraocular pressure (IOP) was (35.0 + or - 0.5) mmHg with a range from 35.0 to 56.0 mmHg. Follow-up periods ranged from 6 to 43 months with a mean of (10 + or - 6) months. Postoperative visual acuity ranged from non light perception to 0.8. Postoperative vision increased in 6 eyes (86%), and was stable in 1 eye (14%). The mean postoperative IOP was (18.0 + or - 1.5) mmHg with a range from 10.0 to 20.0 mmHg, this was significantly lower than that preoperatively (t = 1.94, P < 0.05). Postoperative complications mainly included fibrous exudates in the anterior chamber at early stage after the surgery (7 eyes) and temporary intraocular pressure elevation (1 eye). There was no retinal detachment and atrophia bulbi. CONCLUSION: Vitrectomy combined with silicone oil injection may be a safe and effective method in treating postoperative endophthalmitis following cataract surgery without intraocular lens removal. PMID- 20021879 TI - [Dry pars plana capsulotomy and anterior vitrectomy using 25-gauge vitrectomy system during children cataract surgery]. AB - OBJECTIVE: To describe the initial surgical experience and report the safety and feasibility of dry pars plana capsulotomy and anterior vitrectomy using a 25 gauge vitrectomy system in pediatric cataract surgery. METHODS: This was a prospective case series study. After anterior continuous curvilinear capsulorhexis (ACCC) and phacoemulsification with intraocular lens (IOL) implantation in the capsule for all patients, a pars plana capsulotomy and anterior vitrectomy was performed with a 25-gauge vitrectomy system using a dry technique in 40 eyes of 30 children with cataracts. RESULTS: All procedures were uneventful and all IOLs were implanted in the capsular bag. No intraoperative complications were noted attributable to the small-gauge instruments. The average follow-up period was eight months (range, 4 to 30). Postoperative intraocular pressure remained stable in most cases, except two eyes (5%) with transient hypotony during the first day after surgery. The postoperative inflammatory response was mild. Ultrasound biomicroscopy showed complete healing of the sclerotomy at four weeks after surgery. No posterior synechia, posterior capsule opacification, IOL capture, vitreous prolapse, choroidal detachment or retinal detachment was observed throughout the follow-up period. CONCLUSIONS: Dry pars plana capsulotomy and anterior vitrectomy using a 25-gauge vitrectomy system is safe and effective for the management of posterior lens capsules and anterior vitreous in pediatric cataract surgery. Prospective and comparative study is recommended to evaluate this procedure and traditional method. PMID- 20021880 TI - [Analysis on gene mutations in a Chinese pedigree with autosomal dominant inheritance cataract]. AB - OBJECTIVE: The aim of this study was to reveal the genetic defect of the autosomal dominant inheritance cataract in a Chinese pedigree. METHODS: Case control study. There were 26 individuals investigated with clinical examination in a Chinese four generations pedigree. The genome DNA of the individuals was extracted by the improved NaI method. The exons of six cataract candidate genes in 204 normal controls and 42 senile cataract patients were screened for the mutation by PCR restriction fragment length polymorphism (PCR-RFLP) methods. RESULTS: The phenotype of the cataract was pulverulent nuclear cataract. A novel C/T transition at nucleotide position 827 was identified in the GJA8 gene that led to a serine to phenylalanine change in codon 276. This mutation was not found in 42 senile cataract patients and in 204 controls. Four single nucleotide polymorphisms (SNPs) were also found in a cataract candidate gene in the family members. CONCLUSIONS: A novel GJA8 gene mutation was found in a Chinese autosomal dominant inheritance cataract pedigree. A substitution, C276T in GJA8 gene, was identified as the most likely causative mutation underlying the phenotype of pulverulent nuclear cataract in all affected family members. PMID- 20021881 TI - [Computer construction and analysis of protein models of mutant fibrillin-1 gene in Marfan's syndrome]. AB - OBJECTIVE: Fibrillin-1, the major constituent of extracellular microfibrils, plays an important role in the molecular pathogenesis of Marfan syndrome (MFS, #54700). The aim of this study was to analyze protein models of the mutation of the fibrillin-1 (FBN1) gene on Arg545Cys and Arg1530Cys which have been reported to cause predominant ectopia lentis in Chinese patients. METHODS: We constructed and analyzed the protein models of the mutant FBN1 gene on Arg545Cys and Arg1530Cys. Fibrillin-1 protein structures were predicted by SWISS-MODEL. Models were viewed in Swiss-Pdb Viewer. RESULTS: Computer construction and analysis of protein models of the mutant FBN1 gene revealed that the mutant Arg545Cys FBN1 protein had various changes on protein's secondary structure with an absence of a helix, decreased hydrogen bond distance, different protein surface solvent accessibility and decreased negative electrostatic potential. The mutant Arg1530Cys FBN1 showed lost of hydrogen bonds, different protein surface solvent accessibility and increased negative electrostatic potential. CONCLUSIONS: Protein models of the mutant FBN1 gene shows significant alterations on the protein's secondary structure based on computer construction and analysis technology. This study provides further evidence for the important effect of the mutant FBN1 on the pathogenesis of human ectopia lentis. PMID- 20021882 TI - [Study on soluble and insoluble lens proteins in contusion cataract of rat model]. AB - OBJECTIVE: To study the water soluble and insoluble lens proteins in contusion cataract of rat model, and to study the influence of heat shock (thermotolerance) or quercetin (heat shock protein inhibitor) on the lens proteins. METHODS: Sprague-Dawley (SD) rats were randomized into four groups (6 in each group): (1) A group, control group. (2) B group, contusion group. (3) C group, heat shock group. (4) D group, quercetin group. In contusion group, contusion of eyes was induced by dropping steel ball against the eyeball (20 g ball dropped from 20 cm height for 100 times each week for 5 weeks). In heat shock group, heat shock was induced by raising the body temperature to 40.5 - 41.5 degrees Celsius for 8 min, 2 - 3 h before the eye contusion (as previously described). Heat shock and contusion was given each week for 5 weeks. In quercetin group, rats were fed with quercetin (100 mg/kg weight) 2-3 h before the contusion. Quercetin and contusion was given each week for 5 weeks. Amount of proteins was measured by using Bradford methods. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS PAGE) was used for the separation and analysis of lens proteins. RESULTS: Five weeks after contusion, the mean water soluble protein of heat shock group was 22.71 + or - 1.99, increased significantly than that of the other three groups. The mean insoluble protein of contusion group was 2.60 + or - 0.48, increased than that of the other three groups. From the analysis of SDS-PAGE, we found that there was a high molecular band near 66 KD in opaque lens and the band became more prominent with the growing opacification of the lens. CONCLUSIONS: In contusion cataract, water insoluble lens protein is increased and water soluble lens protein is decreased. SDS-PAGE showed that lens injury resulted in lens proteins moving toward high molecular band. PMID- 20021883 TI - [Clinical research on movement of intraocular contact lens during accommodation]. AB - OBJECTIVE: To study the movement of intraocular contact lens (ICL) during accommodation. METHODS: Forty one eyes of 41 consecutive patients implanted ICL to correct high myopia were collected. The visual acuity and accommodation power were checked before and after the surgery. The movement of both the ICL and the lens was measured separately at nonaccommodated state and pilocarpine induced accommodation. RESULTS: The uncorrected visual acuity (UCVA) and best corrected visual acuity (BCVA) improved 3 months after surgery. The mean accommodation power before surgery was (2.12 + or - 1.06) D, and was (4.46 + or - 2.11) D 3 months after surgery. Accommodation power was improved significantly (t = 2.312, P = 0.022). The distance from the posterior surface of ICL to the anterior surface of the lens was (0.48 + or - 0.27) mm at nonaccommodated state, and (0.34 + or - 0.19) mm at drug induced accommodation, the difference was significant (t = 2.104, P = 0.038). The position from corneal endothelium to the anterior surface of ICL had no significant difference at nonaccommodated state and drug induced accommodation (t = 1.165, P = 0.149). Depth of the anterior chamber (from the posterior surface of the cornea to the anterior surface of the lens) between nonaccommodated and drug-induced accommodation had no statistically significant difference (t = 1.821, P = 0.071), but the anterior pole of the lens actually moved forward at drug induced accommodation checked with slit lamp. With ultrasound biomicroscopy we could see that in several surgical eyes, ICL loop contacted with periphery lens both in nonaccommodated state and drug induced accommodation condition. CONCLUSIONS: Although the constriction of pupil induced by pilocarpine decreases the distance from the posterior surface of ICL to the anterior lens surface significantly, there still has enough space to avoid the occurrence of subcapsular cataract. The contact between ICL loop and periphery lens might be one of the causes for periphery lens opacification, but the influence of this contact to the occurrence of central subcapsular opacification still requires further investigation. PMID- 20021884 TI - [Effects of central corneal thickness and corneal curvature on measurement of intraocular pressure with Goldmann applanation tonometer and non-contact tonometer]. AB - OBJECTIVE: To investigate the effects of central corneal thickness (CCT) and corneal curvature (CC) on intraocular pressure (IOP) measurements by the Goldmann applanation tonometer (GAT) and the non-contact tonometer (NCT). METHODS: One hundred and twenty patients were recruited from the clinic of Peking Union Medical College Hospital. The CCT was measured by ultrasound pachymetry and the mean radius of CC by using Canon PK-5 refractometer. The IOP of each eye was measured by both GAT and NCT. Linear regression was used to compare the measurements of GAT and NCT; multi regression was used to analyze the relationships between CCT, CC and the measurements of GAT and NCT. Bland-Altman method was used to compare the effect of NCT and GAT on the IOP measurements. RESULTS: The results of the right eyes were reported in this paper. The mean and standard deviation of IOP measured by GAT and NCT was (18.4 + or - 4.0) mm Hg (1 mm Hg = 0.133 kPa) and (17.0 + or - 4.6) mm Hg, respectively, the difference was statistically significant (r = 0.835, P = 0.000). IOP measured using GAT increased by 0.039 mm Hg per microm increase in CCT. IOP measured using NCT increased by 0.064 mm Hg per microm increase in CCT. For an increase of 1 mm of mean corneal curvature there was decrease in IOP of 2.648 mm Hg measured by the GAT and of 3.190 mm Hg measured by the NCT. Compared to the GAT, NCT underestimated at low IOP level and overestimated at higher IOP level. CONCLUSIONS: The IOP measurement obtained with both GAT and NCT varied with CCT and CC. CCT affected IOP measurements by NCT more than that by GAT. PMID- 20021885 TI - [Evaluation of serum levels of SOD and MDA in patients with Leber's hereditary optic neuropathy carrying the mitochondrial DNA G11778A mutation]. AB - OBJECTIVE: To determine the serum levels of total superoxide dismutase (SOD) activity and malondialdehyde (MDA), and to evaluate the oxidant-antioxidant status in patients with Leber's hereditary optic neuropathy (LHON) carrying the mitochondrial G11778A mutation. METHODS: Nineteen patients and 12 carriers from three Chinese G11778A LHON families were enrolled in this study, and 30 age matched healthy volunteers were recruited as normal controls. The serum levels of total SOD activity and MDA in all subjects were measured by xanthine oxidase test and thiobarbituric acid technique, respectively. RESULTS: The serum level of total SOD activity in LHON patients was significantly less than that in carriers and normal controls (q = 7.085 and 8.351, respectively, both P < 0.05), however, there was no significant difference between the carriers and normal controls (q = 0.269, P > 0.05). The serum level of MDA in patients and carriers was significantly higher than that in normal controls (q = 9.069 and 4.748, respectively, both P < 0.05), and it was also significantly higher in patients than that in carriers (q = 3.618, P < 0.05). CONCLUSIONS: Antioxidant capacity decreased significantly in patients with LHON, which indicates that the onset of LHON was related with the oxidation-antioxidation imbalance. PMID- 20021886 TI - [Correlations of telomere length changing and pathogeny of keratoconus]. AB - OBJECTIVE: To study telomere length, senescence-associated-beta-galactosidase (SA beta-galactosidase) and senescence marker protein-30 (SMP-30) in the stromal cells of keratoconus or normal corneas respectively, aiming finding the association of these indexes with the phenotype of keratoconus. METHODS: Experiment research. 37 keratoconus lesions corneas were removed from 32 keratoconus patients who were operated in Shangdong Eye Institute between January 2006 and December 2006, and 20 normal corneas were collected from eye bank. The keratoconus corneas ages were from 13 to 34 years [mean ages (19 + or - 5) years] and the control group consists of 20 normal corneas donor ages from 9 to 30 years [mean ages (19 + or - 4) years]. And there was no statistical difference of ages between keratoconus and normal corneas. Southern blot method was utilized to detect telomere length of genomic DNA. SA-beta-galactosidase was detected by 5 bromo-4-chloro-3-indolyl-beta-D-galactopyranoside (X-Gal) staining method respectively in keratoconus and normal corneas. Isolated mRNA from keratoconus and normal corneas were reverse-transcribed to cDNA and SMP-30 was detected using PCR with specific primers (sense: 5' ccg tgg atg cct ttg act at 3'; anti-sense: 5' caa ctt cat gc tgct ttg ga 3'). To compare normal corneas and keratoconus corneas by histopathological study. Statistical analysis by t test. RESULTS: The telomere length in stromal cells in keratoconus corneas were from 10.29 to 14.12 kb, mean (11.54 + or - 1.41) kb, while that of normal corneas were from 12.64 to 15.32 kb, mean (13.45 + or - 0.99) kb. The difference of telomere length in stromal cells of keratoconus and normal corneas reached a statistical significant level (t = 4.753, P < 0.05). That means the telomere length of keratoconus stroma was shorter than that of normal corneal stroma. Light microscopy revealed that collagen fibers in keratoconus corneal stroma were arranged in an irregular manner. Cells density in keratoconus stroma appeared lower than in normal ones but the decrease was not significant. The staining of SA-beta-galactosidase in the keratoconus section was evident, but there was no staining in the normal corneas. SMP-30 was not detectable with RT-PCR method in either keratoconus or normal corneas. CONCLUSION: Telomeres in the keratoconus stromas manifest higher SA-beta-galactosidase than control, implying that improper senescence might be involved in pathogenesis of keratoconus. PMID- 20021888 TI - [Developmental changes of spontaneous excitatory postsynaptic currents mediated by AMPA receptor in layer 2 and 3 pyramidal neurons of rat visual cortex]. AB - OBJECTIVE: To investigate the modification function of visual experience onto synapses by recording developmental changes of spontaneous excitatory postsynaptic currents (sEPSC) in layer 2 and 3 pyramidal neurons of rat visual cortex, and to observe the spontaneous synaptic activities during the earlier postnatal period. METHODS: This was an experimental study. By combining infrared differential interference contrast (IR-DIC) technique and a CCD-camera system with visual patch clamp whole-cell recording technique, spontaneous EPSCs of P2 approximately 7, P8 approximately 14, P15 approximately 21 and P22 approximately 28 groups were observed and analyzed. The neurons were stained by adding 0.3% Lucifer yellow to the pipette solution simultaneously. RESULTS: The amplitude of sEPSC was enhanced when aging (F = 20.69, P < 0.01). The sEPSC frequency increased with age (F = 87.46, P < 0.01). There was no significant difference between P2 approximately 7 groups and P8 approximately 14 groups (P > 0.05). The electrophysiology and dendrite of neuron became mature whilst development. CONCLUSIONS: During the early postnatal days neurons were relatively immature, and became mature during development. Visual experience played an important role in this process. In spite of most synapses were taken silent at early postnatal days, and NMDA receptor-mediated function was revealed exclusively, these results suggested that early AMPA receptor-mediated functional activities existed in layer 2 and 3 pyramidal neurons. Synapses were not completely silent, but had early spontaneous activities. PMID- 20021887 TI - [Comparison of the activities of silver nitrate with those of three antifungal agents against ocular pathogenic fungi in vitro]. AB - OBJECTIVE: To investigate antifungal activity of silver nitrate compared with fluconazole, ketoconazole and amphotericin B against ocular pathogenic fungi in vitro. METHODS: It was an experimental study. Susceptibility tests were performed against 260 isolates (15 genera and 29 species) of ocular pathogenic fungi by broth dilution antifungal susceptibility testing of filamentous fungi (M38-A) approved by National Committee for Clinical Laboratory Standards (NCCLS). Final concentrations ranged from 0.031 to 16.000 mg/L for silver nitrate, ketoconazole and amphotericin B, from 0.5 - 256.0 mg/L for fluconazole. Minimum inhibitory concentration (MIC) was defined as the lowest drug concentration that showed absence of growth or complete growth inhibition (100%). The end points were determined as 100% growth inhibition for silver nitrate and amphotericin B, and > or = 75% growth inhibition for ketoconazole and fluconazole. RESULTS: The MICs at which 90% of isolates were inhibited (MIC(90)) of silver nitrate, ketoconazole, amphotericin B and fluconazole were 2.000, 512.000, 32.000 and 2.000 mg/L for Fusarium species, respectively; 1.000, 256.000, 2.000 and 2.000 mg/L for Aspergillus species, respectively; 2.000, 128.000, 4.000 and 2.000 mg/L for Alternaria alternate, respectively; 2.000, 4.000, 0.125 and 0.500 mg/L for Curvularia lunata, respectively; and 1.000, 256.000, 1.000 and 1.000 mg/L for unusual ocular pathogens, respectively. Silver nitrate was highly active against Aspergillus species (92.9% susceptible at a MIC of < or = 1.0 mg/L) and Fusarium species (96.3% susceptible at a MIC of < or = 2.0 mg/L). 95.6% of Fusarium species and 90.8% of Aspergillus species exhibited resistance to fluconazole, 44.1% of Fusarium species and 42.9% of Aspergillus species exhibited resistance to amphotericin B, 66.2% of Fusarium species exhibited resistance to ketoconazole. The activity of silver nitrate against the fluconazole-resistant, ketoconazole-resistant and amphotericin B-resistant strains was high. CONCLUSION: Silver nitrate has promising activity against a wide variety of ocular pathogenic fungi in vitro, and may have a role in future studies of antifungal eye drops and treating fungal keratitis. PMID- 20021889 TI - [Selective non-peptide CXCR2 antagonist SB225002 inhibits choroidal neovascularization in rat model]. AB - OBJECTIVE: We tested the applicability of using SB225002, a selective non-peptide CXCR2 inhibitor, for inhibiting experimental choroidal neovascularization (CNV) in Brown Norway (BN) rats. METHODS: It was an experimental study. CNV was induced in 12 adult BN rats with laser photocoagulation. 10 micromol/L SB225002 was administered into the vitreous of 6 rats right after laser injury. DMSO was used as the control in other 6 rats. Three BN rats were served as non-laser-treated controls in quantitative real-time reverse transcription PCR (qRT-PCR) analysis. Fluorescein angiography was performed on day 7 postoperatively to observe the fluorescein leakage of CNV. Quantitative analysis on choroidal flat mounts was performed to evaluate the area changes of CNV lesions. qRT-PCR analysis was used to compare the changes shown by the SB225002-, DMSO- and non-laser-treated BN rats with regard to mRNA levels of CXCR2 and vascular endothelial growth factor (VEGF) in the RPE-choroidal complex. RESULTS: SB225002 (10 micromol/L) significantly inhibited the fluorescein leakage (P < 0.05), the extent of neovascularization on choroidal flat mount of rat CNV model was decreased up to 66% (t = 2.54, P = 0.001). CXCR2 and VEGF mRNA were reduced significantly following this treatment (t = 8.54, 3.61; P = 0.007, 0.002). CONCLUSION: SB225002 inhibits angiogenic activity of IL-8 by blocking its binding with CXCR2 in the early stage of CNV, which may provide a potential new therapeutic strategy for CNV. PMID- 20021890 TI - [Small interference RNA targeting vascular endothelial growth factor inhibits rat corneal neovascularization]. AB - OBJECTIVE: To test the effects of VEGF-siRNA-transfected corneal epithelium on corneal neovascularization (CNV). METHODS: It was an experimental study. Cultured rat corneal epithelial cells and keratocytes were transfected with synthesize VEGF siRNA by lipofectamine 2000. The level of VEGF mRNA was analyzed by real time PCR, and the protein levels were determined by enzyme-linked immunosorbent assay (ELISA). CNV was induced by cauterization with 1 mol/L sodium hydroxide in rat corneas. The VEGF-siRNA-transfected-corneal epithelium cells were transplanted to the CNV lesions. Immediately after transplantation, the VEGF siRNA combined with lipofectamine 2000 were directly transfected rat cornea through injecting into the anterior chamber. After surgery, the surface areas occupied by new vessels were measured, and VEGF protein was localized by immunohistochemistry. RESULTS: The levels of VEGF expression at both mRNA and protein in the VEGF-siRNA transfected corneal epithelial cells and keratocytes were significantly lower than those of control cells. VEGF siRNA could inhibit the expression of VEGF mRNA in corneal epithelial cells and keratocytes to 57% - 85% and 59% - 78%, respectively. The VEGF-siRNA-transfected-corneal epithelium transplantation significantly decreased the surface areas occupied by new vessels. VEGF expression level in interference groups was lower than that in the control group. CONCLUSIONS: The development of CNV is markedly suppressed by VEGF siRNA transfection in vivo. PMID- 20021895 TI - [Dry transconjunctival sutureless 25-gauge vitrectomy in the treatment of pediatric cataract]. AB - Posterior capsule opacification is the most frequent complication of pediatric cataract surgery. To prevent posterior capsule opacification, primary phacoemulsification, posterior capsulotomy and anterior vitrectomy with intraocular lens implantation is the preferred method in the treatment of pediatric cataract. Anterior vitrectomy cutter, with 18-gauge, maximum frequency at 600/min and has simultaneous cutting, irrigation and aspiration functions, is associated with more complications and poor outcomes. In 20-gauge surgery, pars plana vitrectomy is performed with two-port sclerotomy. The irrigation increases movement of vitreous and 20-gauge sclerotomy needs suture for closing. In 25 gauge surgery, the vitreous cutter can be introduced into the vitreous cavity directly though conjunctiva and sclera. The stab incision is roughly half the size of 20-gauge cutter, therefore, the sclerotomy incision can be left unsutured. Surgery with dry transconjunctival sutureless 25-gauge vitrectomy may decrease the requirement for secondary membrane surgery and the risk for retinal detachment. The application of dry transconjunctival sutureless 25-gauge vitrectomy in the treatment of pediatric cataract is reviewed. PMID- 20021897 TI - [To further improve the effects of surgical treatment for hilar cholangiocarcinoma]. PMID- 20021898 TI - [Criteria for radical resection of hilar cholangiocarcinoma]. PMID- 20021899 TI - [Current situation and evaluation of surgical treatment of hilar cholangiocarcinoma]. PMID- 20021900 TI - [Dispute issues related to surgical treatment of hilar cholangiocarcinoma]. PMID- 20021901 TI - [The impact of preoperative biliary drainage on surgical morbidity in hilar cholangiocarcinoma patients]. AB - OBJECTIVE: To evaluate the impact of preoperative biliary drainage on surgical morbidity in hilar cholangiocarcinoma patients underwent surgery. METHODS: One hundred and eleven consecutive patients with hilar cholangiocarcinoma whose serum total bilirubin (TBIL) level > 85 micromol/L and underwent surgery in the period from June 1998 to August 2007 were enrolled. There were 67 male and 44 female patients, aged from 26 to 82 years old with a mean of 56 years old. RESULTS: Fifty-five patients underwent preoperative biliary drainage with a mean of 11.4 d of drainage period (drainage group), the other (n = 56) were the non-drainage group. The preoperative TBIL level of drainage group was (154 +/- 69) micromol/L, which was significantly lower than the value of pre-drainage (256 +/- 136) micromol/L (P = 0.000) and the value of non-drainage group (268 +/- 174) micromol/L (P = 0.005). ALT and GGT levels could be lowered by preoperative biliary drainage. The postoperative complications of these two groups were comparable (36.3% vs. 28.6%, P = 0.381). Four patients in drainage group and 5 patients in non-drainage group died of liver failure. Multivariate logistic regression indicated that hepatectomy (OR = 0.284, P = 0.003) was the independent risk factor associated with postoperative morbidity. Bismuth-Corlette classification (OR = 0.211, P = 0.028) was the independent risk factor linked to postoperative mortality. CONCLUSIONS: Preoperative biliary drainage could alleviate liver injury due to hyperbilirubin, but it could not decrease the surgical morbidity and postoperative mortality. Concomitant hepatectomy and Bismuth-Corlette classification were independent risk factors linked to surgical risks. PMID- 20021902 TI - [Experience from surgical resection for 48 cases of hilar cholangiocarcinoma]. AB - OBJECTIVE: To summarize the methods, safety and efficiency of surgical resection for hilar cholangiocarcinoma. METHODS: The clinical and follow-up data of 48 patients with hilar cholangiocarcinoma underwent surgical resection from January 2003 to December 2007 were analyzed retrospectively. There were 26 male and 22 female, aged from 38 to 72 years old with a mean of 63.6 years old. RESULTS: Perioperative management including percutaneous transhepatic biliary drainage applied in 19 cases and portal vein embolization applied in 2 cases. Eight patients were treated with extrahepatic bile duct resection with or without parital hepatic segment II resection, 10 cases with perihilar hepatic resection (segment IVB, partial V, partial VIII, I), 28 cases with extended hemihepatectomy and 2 cases with central hepatic resection (segment IVB, V, VIII, I). R0 resection rate was 89.5% and the operative mortality was 2.1%. The 1-, 3- and 5 year survival rate were 93.5%, 51.8% and 36.5%, respectively. Patients undergoing extended hepatic resection survived significantly longer than those undergoing partial hepatic resection (P = 0.034). CONCLUSIONS: Extended hepatic resection for hilar cholangiocarcinoma offers good outcomes with an acceptable mortality rate. PMID- 20021903 TI - [Surgical treatment of hilar cholangiocarcinoma:experience of 53 cases]. AB - OBJECTIVE: To summarize the experience of surgical treatment of the hilar cholangiocarcinoma and explore the factors influencing the operative effect. METHODS: Fifty-three cases of hilar cholangiocarcinoma performed operation from January 1998 to January 2008 were divided into two groups: Group I included cases treated from January 1998 to December 2002 (totally 22 cases); Group II included cases treated from January 2003 to January 2008 (totally 31 cases). Comparative study was carried out between these two groups. RESULTS: Surgical resection [radical resection (R(0)) + palliative resection (R(1))] rate in group I was lower than that in group II (40.9% vs. 71.0%, P = 0.029). Hepatic lobectomy rate in group I was lower than that in group II (30.0% vs. 64.5%, P = 0.016). The resection rate was positively correlated with the hepatic lobectomy rate (r = 0.985, P < 0.01). The median survival time of the patients underwent R(0) resection was longer than that in the patients underwent R(1) resection and palliative bile duct drainage [(27.0 +/- 4.5) months vs. (10.0 +/- 0.7) months vs. (4.0 +/- 0.5) months, respectively, P < 0.01]. CONCLUSIONS: Early diagnosis and radical resection are important to improve the prognosis for patients with hilar cholangiocarcinoma. But for the patients could only receive palliative resection or with positive celiac trunk lymph nodes, combined liver lobe resection could not improve the survival. PMID- 20021904 TI - [Experience of surgical resection of Bismuth-Corlette type I and II hilar cholangiocarcinoma]. AB - OBJECTIVE: To report the experience of surgical resection of Bismuth-Corlette type I and II hilar cholangiocarcinoma. METHODS: From January 1998 and January 2008, 52 cases of Bismuth-Corlette type I and II hilar cholangiocarcinoma were operated on. The clinical data and long-term outcome of the patients was retrospectively analyzed. RESULTS: Of the 52 cases, 44 cases (84.6%) received operation, 28 patients underwent radical resection (63.6%) and 16 patients (36.4%) underwent palliative resection.Seven patients were resected on caudate lobe and other section and lobe of the liver; among them, 2 patients received combined portal vein resection and 4 underwent combined hepatic artery resection respectively. Eleven cases developed postoperative complications and another one died in hospital. The median survival was 33.2 months in radical resection group, and 1-, 3-, 5-year survival rate was 82.6%, 47.8%, 34.7%, respectively, which was significant greater than those in the palliative resection group (41.6%, 16.6%, 8.3%, respectively) (P < 0.05). The median survival was 16.7 months in the palliative resection group. CONCLUSIONS: The radical resection is still the best treatment for Bismuth-Corlette type I and II hilar cholangiocarcinoma. Intraoperative pathology for resection margin, and combined liver resection, portal vein resection and hepatic artery resection can help improve the radical resection rate. PMID- 20021905 TI - [Minor liver resection for hilar cholangiocarcinoma of Bismuth-Corlette type III]. AB - OBJECTIVE: To evaluate long-term outcomes of minor liver resection for hilar cholangiocarcinoma (HC) of Bismuth-Corlette type III. METHODS: From January 1997 to December 2007, the clinical data of 91 patients with Bismuth-Corlette type III HC underwent hepatectomy were collected and analyzed retrospectively. RESULTS: There were 60 patients underwent minor hepatectomy, and 31 undergoing major hepatectomy. Hepaticojejunostomy was made conventionally in an end-to-side fashion in the patients undergoing major liver resection, and a new technique of hepaticojejunostomy used in the patients undergoing minor liver resection. That was the anterior edges of bile duct stumps which were not sutured after suturing of posterior edges. Instead of, the anterior edge of jejunum loop to the remnant liver on the top of the bile duct stumps were sutured with intermittent "U" sutures. In all patients, in-hospital mortality rate was 0 and rate of bile leakage was only 2.1%. The actual 1-, 3- and 5-year survival rates were 91.6% and 87.0%, 61.6% and 62.0%, 31.6% and 33.0%, respectively (P > 0.05). CONCLUSIONS: Minor liver resection for the selected patients with HC of Bismuth-Corlette type III according to our criteria achieved better long-term outcomes. A new hepaticojejunostomy used in the patients undergoing minor liver resection is a safe and effective method. PMID- 20021906 TI - [Surgical treatment for hilar cholangiocarcinoma of Bismuth-Corlette type IV]. AB - OBJECTIVE: To investigate the surgical treatment for hilar cholangiocarcinoma(HCCA) of Bismuth-Corlette type IV. METHODS: The clinical data of 73 HCCA patients admitted in Southwest Hospital, the Third Military Medical University from January 2002 to December 2008 were analyzed retrospectively. There were 41 males and 32 females, aged from 30 to 84 years old (averaged, 56.8 years old). All patients were diagnosed as hilar mass with hilar biliary obstruction by B-ultrasound, CT, magnetic resonance imaging, endoscopic retrograde cholangiopancreatography, magnetic resonance cholangiopancreatography or percutaneous transhepatic cholangiography, and confirmed by pathological examination in intra-operation and post-operation. Diagnosis of all patients were according to Bismuth-Corlette type IV. RESULTS: Fifteen cases underwent percutaneous transhepatic cholangial drainage, stents were implanted in 8 patients. Simple internal drainage was performed on 25 of the 73 patients and 4 with palliative resection. Radical resection was performed on 19 patients. The radical resection rate of HCCA were 26.0%. The 1, 3 years of survival rates were 36.8%, 10.5% respectively. The 1 year survival rate was 6.3% in drainage group. CONCLUSION: Radical resection is the potentially curative treatment for HCCA, a sufficient, reasonable use of internal and external drainage would improve the patient's quality of life. PMID- 20021907 TI - [Long-term, tumor-free survival after radiotherapy combining hepatectomy-Whipple en bloc and orthotopic liver transplantation for early-stage hilar cholangiocarcinoma]. AB - OBJECTIVE: To report the experience in surveillance and early detection of cholangiocarcinoma (CC) and in using en bloc total hepatectomy pancreaticoduodenectomy-orthotopic liver transplantation (OLT-Whipple) to achieve complete eradication of early-stage CC complicating primary sclerosing cholangitis (PSC). METHODS: Asymptomatic PSC patients underwent surveillance using endoscopic ultrasound and endoscopic retrograde cholangiopancreatography (ERCP) with multilevel brushings for cytological evaluation. Patients diagnosed with CC were treated with combined extra-beam radiotherapy, lesion-focused brachytherapy, and OLT-Whipple. RESULTS: Between January 1988 and February 2001, 42 of 119 PSC patients were followed according to the surveillance protocol. CC was detected in 8 patients, 6 of whom underwent OLT-Whipple. Of those 6 patients, 4 had stage I CC, and 2 had stage II CC. All 6 OLT-Whipple patients received combined external-beam and brachytherapy radiotherapy. The median time from diagnosis to OLT-Whipple was 144 days. One patient died 55 months post-transplant of an unrelated cause, without tumor recurrence. The other 5 were well without recurrence at 79, 82, 108, 128, 129 and 145 months. CONCLUSIONS: For patients with PSC, ERCP surveillance cytology and intralumenal endoscopic ultrasound examination allow for early detection of CC. Broad and lesion-focused radiotherapy combined with OLT-Whipple to remove the biliary epithelium en bloc offers promising long-term, tumor-free survival. All patients tolerated this extensive surgery well with good quality of life following surgery and recovery. These findings support consideration of the complete excision of an intact biliary tree via OLT-Whipple in patients with early-stage hilar CC complicating PSC. PMID- 20021908 TI - [Biopathological characteristics of hilar cholangiocarcinoma and the clinical significance]. AB - OBJECTIVE: To approach the biopathological features of hilar cholangiocarcinoma and surgical pathological factors which influence the long-term survivals of patients with hilar cholangiocarcinoma. METHODS: A systemic and retrospective multi-parameter analysis was performed on 205 patients of hilar cholangiocarcinoma who received surgical treatments and had complete clinicopathological data as well as follow-up results during a ten-year-period from April 1998 to April 2008. The single factor analysis was performed on age, sex, content of pre-operative serum CA19-9, Child-pugh grading, TNM classification, operation pattern, resection margin status of bile duct, vascular invasion, adjacent liver involvement, grade differentiation, infiltration-depth of bile duct, lymph node metastasis and perineural infiltration. A multivariate analysis was performed through Cox proportional hazard model. RESULTS: The single factor analysis showed that except age, sex and content of pre-operative serum CA19-9, the mainly significant factors influencing the survivals were Child-Pugh grading, TNM classification, operation pattern, bile duct margin, vascular invasion, adjacent liver involvement, grade differentiation, infiltrating-depth of bile duct, lymph node metastasis and perineural infiltration (P < 0.05). Lymph node metastasis and infiltration-depth of bile duct wall were found to be the two independent factors influencing overall survival by multivariate analysis through the Cox model. CONCLUSIONS: The most important prognostic factors influencing the long-term survivals of patients with hilar cholangiocarcinoma after operation are lymph node metastasis and depth of tumor-infiltrating of involved bile duct. During the operation, standardized evaluation through frozen section should be carried out for detection of lymph node metastasis and depth of tumor infiltrating of involved bile ducts, which can be used as the histological indicator for surgical expansion, and could be helpful to maximize avoiding the tumor cell residues and therefore, to improve the long-term effects of surgical resection. PMID- 20021909 TI - [Anatomy problems in the radical resection for hilar cholangiocarcinoma]. AB - OBJECTIVE: To analyze the clinical data, surgical strategies and results from the patients with hilar cholangiocarcinoma (HCCA), and to explore the anatomic factors related to the radical resection. METHODS: The data from 52 patients with HCCA who underwent radical resection between January 1984 to December 2008 were investigated retrospectively, which included clinical diagnosis, Bismuth-Corlette classification, pathologic features, surgical procedures and follow-up results. RESULTS: According to the Bismuth-Corlette classification, 5, 12, 6, 16 and 13 patients belonged to type I, II, IIIa, IIIb and IV respectively. There were 24 cases underwent combined hepatic lobectomy. The 1-, 3- and 5-year survival rates were 78.8%, 36.4% and 12.1% respectively. Postoperative complications rate was 30.8% with the 3.8% mortality rate. The frequency of surgical complications was significantly higher in patients with higher level of serum total bilirubin (> 340 micromol/L) than that in patients with a relatively lower one (170 micromol/L) before operation (P < 0.05). CONCLUSIONS: Some anatomical factors should be considered during the radical resection of hilar cholangiocarcinoma, especially evaluation of potential hepatectomy, resection of caudate lobe, hepatic artery resection and/or reconstruction. The prognosis of the patients underwent R(0) radial resection could be significantly improved. PMID- 20021910 TI - [Neoadjuvant chemotherapy of FLEEOX regimen for unresectable gastric cancer resulting from advanced abdominal lymph nodes metastases]. AB - OBJECTIVE: To observe the therapeutic response of advanced gastric cancer with severe lymph nodes metastasis to FLEEOX regimen neoadjuvant chemotherapy that combined arterial and venous administration. METHODS: Neoadjuvant chemotherapy was administered to 32 cases of gastric cancer with advanced abdominal lymph nodes metastases from January 2007 to October 2008. Of the 32 patients, 28 had severe local lymph nodes metastasis, such as No.3, 7, 9, 12 lymph nodes metastasis, one patient had No.16 lymph nodes metastasis, and the other 3 patients had both regional and No.16 lymph nodes metastasis under CT scan. Neoadjuvant chemotherapy was administered as follows: 5-Fu 370 mg/m(2), intravenous drip, day 1 - 5; Leukovorin 120 mg, intravenous drip, day 1 - 5; oxaliplatin 150 mg/m(2), epirubicin 30 mg/m(2) and epotoside 70 mg/m(2), intravascular infusion through arteria gastrica sinistra, day 6 and 20. The protocol was repeated every five weeks for two or three courses. After 2 or 3 cycles of chemotherapy, abdominal CT was taken to evaluate the radiological therapeutic response and calculate the reductive rate of the tumor. RESULTS: The general conditions and symptoms was improved significantly in all the patients. Four cases got complete response (CR), 24 got partial response (PR) and 4 got no change under CT scan. For the 32 case, the radiological response rate (CR + PR) was 87.5%(28/32). Thirty patients underwent subtotal or total gastrectomy, even combined organ resection, with D2 or D2 + alpha lymphadenectomy. CONCLUSIONS: The FLEEOX regimen which combines arterial and venous administration carries a satisfactory therapeutic effect in advanced gastric cancer with severe lymph nodes metastasis. The combined routes of drug administration may improve the effects of neoadjuvant chemotherapy in stomach cancer. PMID- 20021911 TI - [The analysis of the follow-up results for the crural artery bypass]. AB - OBJECTIVE: To review the follow-up results of the crural artery bypass. METHODS: Sixty-five limbs in 64 patients with long stenosis or occlusion in femoral artery and popliteal artery were performed 65 times femoral-crural artery bypass surgery or femoral-popliteal-crural bypass surgery during April 2001 to July 2007. The ankle-brachial index before bypass surgery was 0.35 +/- 0.20 in anterior tibial artery and 0.38 +/- 0.21 in posterior tibial artery. Critical limb ischemia was 93.8%. RESULTS: The ankle-brachial index after bypass surgery was 0.84 +/- 0.26 in anterior tibial artery and 0.83 +/- 0.22 in posterior tibial artery. The perioperative mortality rate was 1.6%, the perioperative amputation rate was 1.5%. Fifty-four patients 54 limbs were followed up. The average follow-up time was (24.1 +/- 16.6) months. The follow-up limb salvage rate was 85.2%. The follow up mortality rate was 25.9%. Critical limb ischemia decreased as 13.0%. The follow-up ankle-brachial index was difference with before and after bypass surgery as 0.66 +/- 0.26 in anterior tibial artery and 0.64 +/- 0.25 in posterior tibial artery. It was no difference in cumulative limb salvage rate, cumulative primary and secondary patency rate by comparing autogenous vein with composite vascular as graft and comparing femoral-crural artery bypass surgery with femoral popliteal-crural bypass surgery as surgical method. CONCLUSIONS: When the patients are failed in endovascular intervention or have long stenosis or occlusion in femoral artery and popliteal artery to face to amputation, the crural artery bypass is a feasible method. It's helpful to improve the secondary patency rate and limb salvage rate by enhancing the follow-up after operation and early intervention. PMID- 20021912 TI - [The relationship between rhGH and blood sugar on different ages of severe degree burned patients]. AB - OBJECTIVE: Probe the effects of rhGH on severe degree burned patients' blood sugar in different age of years. METHODS: Elected 210 patients hospitalized in the Third Affiliated Hospital of Wenzhou Medical College from January 2005 to December 2008, who were burned in 48 h, older than 18 years, ever had no diabetes and tumor history and placidly pull through shock stage. Among the patients there were 132 males and 78 females. The age was from 18 to 65 years old, average (40.7 +/- 7.2) years old. The extent of burn were form TBSA 25% to TBSA 86%, average TBSA (40.4 +/- 12.5)%. The depths of burn were from superficial second degree to third degree. All of the total divided into A (18 - 44 years old) and B (> 45 years old)groups. Each group had 105 patients. Two groups were randomly divided into A(1), A(2), A(0) and B(1), B(2), B(0) groups. Each group had 35 patients. The A(1) and B(1) groups were used 0.15 U/(kg.d) growth hormone (Somatropin, S19990021), A(2) and B(2) groups were used 0.2 U/(kg.d) growth hormone, A(0) and B(0) groups were used NS as control. Observed and analyzed the change of blood sugar and insulin amount used in 210 patients. RESULTS: Of all the patients in 6 groups, there were 190 patients finished the experimentation in four weeks. The insulin amount of A(1), A(2), A(0) groups used were (2123.3 +/- 152.3), (2885.6 +/- 148.5), (724.1 +/- 31.1) U, B(1), B(2), B(0) group were (2715.1 +/- 95.3), (3652.2 +/- 198.1), (801.8 +/- 22.2) U. The consequence showed that the number need insulin to control blood sugar in B group was more than A group, as well as using 0.2 U/(kg.d) does to 0.15 U/(kg.d) does, and using growth hormone to no using(P < 0.01). The time that blood sugar of A(1), A(2), B(1), B(2) group recovered to normal range without using insulin were (5.11 +/- 0.82), (4.93 +/- 0.89), (5.2 +/- 0.65), (5.13 +/- 1.02) d (P > 0.05). CONCLUSIONS: The blood sugar's alteration has positive correlation with the age of years and the does of rhGH. As long as normative using rhGH it doesn't induce diabetes. PMID- 20021913 TI - [Clinical anatomic study of internal fixation of acetabular posterior column plate technique]. AB - OBJECTIVES: To study the best entry points, direction and length of screw in acetabular posterior column plate technique, and to prevent the serious complications of screw penetrating the joint surface. METHODS: February to July 2008 20 male cadaveric adult semi-pelvic specimen were taken.Serial cross sections of the acetabular posterior column were determined and made. The safe angle of screw entry and the length on all entry points of each cross-section were measured. And put all data into software SPSS 10.0 for statistics process. RESULTS: On margin of acetabulum, lateral-middle 1/4 point, midpoint, medial middle 1/4 point and medial margin of posterior column of each cross-section, safe entry angle of inclination was 39 degrees , 57 degrees , 74 degrees , 90 degrees and 106 degrees respectively, the length of the screw was 39, 57, 74, 90 and 106 mm respectively. CONCLUSION: On lateral 1/4 region, lateral-middle 1/4 region, medial-middle 1/4 region and medial 1/4 region, screw posterior column angle is 40 degrees to 60 degrees , 60 degrees to 75 degrees , 75 degrees to 90 degrees , 90 degrees to parallel to the quadrilateral plate, and the length of the screw is 30 mm. PMID- 20021914 TI - [Diabetes counteracts the protective effect of the diazoxide preconditioning on ischemic reperfused rat heart]. AB - OBJECTIVE: To explore the impact of diabetic condition on the protective effect of diazoxide preconditioning (DPC) on ischemic-reperfused (I/R) myocardium in rats. METHODS: Thirty normal male Sprague-Dawley rats were divided into 3 groups, including non-diabetic control group, non-diabetic I/R group, and non-diabetic I/R DPC group. Thirty diabetic male rats were also divided into the same 3 groups. The Langendorff isolated heart perfusion models were established. The control groups had a 90 min perfusion without any intervention. The I/R groups had a 30 min equilibration period, a 30 min ischemia, and a 30 min reperfusion. The I/R DPC groups had a 10 min equilibration, two cycles of 100 micromol/L diazoxide perfusion, 5 min each, followed by a 5 min diazoxide-free period before the 30 min ischemia and a 30 min reperfusion. The recovery rate of the left ventricular function, including cardiac output, left ventricular developed pressure (LVDP), and the maximum change rate of left ventricular pressure rise and fall (+/- dp/dt(max)) were recorded. The activity of creatine kinase in coronary outflow and activities of malonyldialdehyde, and superoxide dismutase in myocardium were detected. Myocardial water content was also assessed. RESULTS: In non-diabetic rats, the content of creatine kinase, malonyldialdehyde and water content were significantly decreased in I/R DPC group compared with those in I/R group. Furthermore, in I/R DPC group, the activity of superoxide dismutase and the recovery rate of the left ventricular function, including cardiac output, LVDP and +/- dp/dt(max), were significantly increased compared with those in I/R group (P < 0.05). By contrast, there were no significant changes between I/R DPC group and I/R group in diabetic rats (P > 0.05). CONCLUSION: Diabetes counteracts the protective effect of the diazoxide preconditioning on ischemic reperfused rat heart, which may be related with acute insulin resistance in cardiomyocytes. PMID- 20021915 TI - [Improving microenvironment is the important approach to enhance stem cell transplantation effect]. PMID- 20021916 TI - [Improve efficacy of stem cell therapy for coronary heart diseases: choosing "seed cells"]. PMID- 20021917 TI - [Efficacy comparison between transplanting microenvironmental induced and non induced bone marrow mesenchymal stem cells in ischemic rat hearts]. AB - OBJECTIVE: To compare the efficacy of transplanting bone marrow mesenchymal stem cell (BMSC) or microenvironmental induced BMSC (iBMSC) into the ischemic myocardium of rats with myocardial infarction. METHODS: iBMSC was defined as BMSC co-cultured with myocardial cells for 2 weeks. The stem cells or equal volume PBS were injected into ischemic border zone 1 wk after experimental infarction. Cardiac performance was evaluated at 1, 2, and 4 wk after cell transplantation by echocardiography and analyzed histologically at 4 wk after cell transplantations. RESULTS: Compared with PBS group, both BMSC and iBMSC transplantations reduced infarct size. iBMSC enhanced the beneficial effects of BMSC on improving cardiac function (FS: 28.5% +/- 4.3% in PBS, 29.0% +/- 2.0% in BMSC and 45.1% +/- 3.1% in iBMSC group at 4 weeks post transplantation, iBMSC group vs. PBS group P < 0.05, iBMSC group vs. BMSC group P < 0.05). Immunofluorescence microscopy results revealed co-localization of SPIO-labeled transplanted cells with cardiac markers for cardiomyocytes, indicating regeneration of damaged myocardium. CONCLUSION: Our data suggest that iBMSC implantation is more effective on improving cardiac function than BMSC implantation in this model. iBMSC might serve as a new promising therapeutic cell source for regenerating ischemic myocardium in patients with post-infarction heart failure. PMID- 20021918 TI - [Intracoronary and hypodermic injection of granulocyte colony-stimulating factor improved cardiac function in Swine with chronic myocardial ischemia]. AB - OBJECTIVES: To compare the efficacy and feasibility between intracoronary and hypodermic injection of granulocyte colony-stimulating factor (G-CSF) on improving cardiac function in a Swine model of chronic myocardial ischemia. METHODS: Eighteen Swine underwent placement of ameroid constrictor on left circumflex coronary artery. The presence of myocardial ischemia was verified at four weeks after the operation, and the animals were then randomly assigned into three groups (n = 6 each): (1) administration of vehicle (control), (2) hypodermic injection of G-CSF (5 microgxkg(-1)x;d(-1)) for five days (IH), and (3) intracoronary injection of a bonus G-CSF (60 microg/kg) (IC). Coronary angiogram, cardiac MRI, and (18)F-FDG-SPECT/(99m)Tc-SPECT (DISA-SPECT) measurements were performed at pre-administration and at 4 weeks post administration. Global heart function such as left ventricular end-diastolic volume (LVEDV), left ventricular end-systolic volume (LVSDV) and left ventricular ejection fraction (LVEF), myocardial perfusion, myocardial viability and myocardial infarct area were evaluated. Myocardial vWF, Bcl-2 and Bax expressions were detected by Western blot and RT-PCR. RESULTS: MRI data showed that left ventricular dilation and dysfunction were similarly prevented in IH and IC G-CSF treated animals at eight weeks after the operation. SPECT revealed that both IH and IC G-CSF equally improved the regional contractility of chronic myocardial ischemia and increased myocardial viability. Myocardial infarct size was also reduced after both G-CSF treatments as detected by MRI. Intracoronary injection of G-CSF did not lead to angiogenesis in other organs. G-CSF treatments were also associated with a significant reduction in myocardial apoptosis and significant increase in angiogenesis. CONCLUSIONS: Both intracoronary and hypodermic injection of G-CSF were safe and feasible and could equally improve cardiac function and increase angiogenesis in this Swine model of chronic myocardial ischemia. PMID- 20021919 TI - [Effects of autologous mesenchymal stem cells transfected with heme oxygenase-1 gene transplantation on ischemic Swine hearts]. AB - OBJECTIVE: To observe the effect of intracoronary transfer of autologous HO-1 overexpressed MSCs in porcine model of myocardial ischemia (1 h)/reperfusion. METHODS: Apoptosis was assayed and cytokine concentrations in supernatant were measured in cells exposed to hypoxia-reoxygen in vitro. In vivo, Chinese male mini-pigs were allocated to the following treatment groups: control group (saline), MSCs group (MSCs), MSCs transfected with pcDNA3.1-nHO-1 (HO-1-MSCs). 1 x 10(7) of autologous stem cells or identical volume of saline was injected intracoronary into porcine hearts 1 h after ischemia. MRI assay and postmortem analysis were assessed 3 months after stem cell transplantation. RESULTS: In vitro, cell apoptosis rate post hypoxia-reoxygen was significantly reduced in HO 1-MSCs group (30.30% +/- 7.64%) compared with that in MSCs group (56.93% +/- 4.68%, P < 0.001) and LacZ-MSCs group (55.88% +/- 4.38%, P < 0.001), VEGF was also significantly upregulated in HO-1-MSCs group [(768.44 +/- 78.38) pg/ml] compared with that in MSCs group [(555.27 +/- 67.67) pg/ml, P < 0.001] and LacZ MSCs group [(522.97 +/- 71.45) pg/ml, P < 0.001]. In vivo, cardiac function was significantly improved in both MSCs transplantation groups compared to saline group (all P < 0.05 vs.saline) and the left ventricular ejection fraction was significantly higher in HO-1-MSCs group compared with that in MSCs group at 3 months after transplantation (53.50% +/- 2.09% vs. 49.54% +/- 2.74%, P = 0.017), capillary density in the peri-infarct area was also significantly higher in HO-1 MSC group than that in MSCs group [(14.59 +/- 2.39)/HPF vs. (11.78 +/- 2.48)/HPF, P = 0.033]. CONCLUSIONS: Efficacy of HO-1 overexpressed MSCs on improving cardiac function and promoting angiogenesis was greater than those by MSCs in this porcine ischemia/reperfusion model. PMID- 20021921 TI - [Effects of amlodipine plus telmisartan or amlodipine plus amiloride regimen on blood pressure control in hypertensive patients: preliminary report of Chinese Hypertension Intervention Efficacy (CHIEF) trial]. AB - OBJECTIVE: To evaluate the effects of amlodipine-based antihypertensive combination regimen on blood pressure control and impact on cardiovascular events. METHODS: From Oct. 2007 to Oct. 2008, a total of 13 542 hypertensive patients from 180 centers in China were included in this multi-centre randomized, controlled, blind-endpoint assessment clinical trial. Inclusion criteria were: essential hypertension, 50 - 79 years of age with at least one cardiovascular risk factor and signed consent forms. Patients were randomly assigned to receive low-dose amlodipine + diuretics (group A) or low-dose amlodipine + telmisartan (group T). The primary endpoints are composite of non-fatal stroke/myocardial infarction and cardiovascular death. All patients will be followed-up for 4 years. RESULTS: The characteristics of patients between the two groups were similar: mean age (61.5 +/- 7.7) Yrs with 19% history of cerebrovascular diseases, 12% coronary diseases, 18% diabetes, 42% dyslipidemia, mean initial blood pressure 157/93 mm Hg. After 8-week treatment, mean blood pressure in group A and B were reduced to (133.0 +/- 11.0)/(81.0 +/- 7.6) mm Hg, (132.9 +/- 11.6)/(80.6 +/- 7.9) mm Hg respectively. Blood pressure control rates reached 72.1% and 72.6% in group A and T, respectively. CONCLUSION: Amlodipine-based antihypertensive combination regimens achieved satisfactory blood pressure control rate in patients with essential hypertension in this patient cohort. PMID- 20021922 TI - [The prognosis study of 108 idiopathic pulmonary arterial hypertension patients]. AB - OBJECTIVE: To analyze the characteristics of idiopathic pulmonary arterial hypertension (IPAH) and predict the prognosis of IPAH by Echocardiogram and Right Heart Catheterization. METHODS: 108 IPAH patients who was diagnosed as IPAH in Fuwai hospital from Jan 2003 to Jun 2007 were retrospectively analyzed and followed up to June 2008, the statistical analysis software was SPSS 13.0. RESULT: The mean age of the 108 IPAH patients was (32.5 +/- 12.6) years, the ratio of female and male was 2.86:1, the average time from appearing symptom to diagnosis was (34.9 +/- 36.3) months, the incidence of syncope, hepatomegaly, hemoptysis and Edema of lower extremity was 32.4%, 27.8%, 15.7% and 14.8% respectively. A total of 94 patients (87.0%) were followed up for (27.6 +/- 14.7) months, forty-two patients (38.9%) died of right heart failure, the mean death time was (11.6 +/- 9.4) months after diagnosis, the survival rate of 1, 2 and 3 years was 74%, 58% and 44% respectively. These following variables could predict survival in univariate analysis: Systolic Blood Pressure (SBP) (P < 0.01, RR = 0.937), pulse Pressure (PP) (P < 0.01, RR = 0.930), left ventricular End Diastolic Dimension (LVEDD) (P = 0.004, RR = 0.924), right ventricular End Diastolic Dimension (RVEDD) (P = 0.029, RR = 1.045), RVEDD/LVEDD (P = 0.003, RR = 3.222), Pulmonary Arterial Systolic Pressure (PASP) measured by echocardiogram (P = 0.002, RR = 1.022), PASP (P = 0.030, RR = 1.026), mean Pulmonary Arterial Pressure (mPAP) (P = 0.031, RR = 1.037) and Pulmonary Vascular Resistance (PVR) measured by right heart catheter (P = 0.019, RR = 1.001). in multivariate analysis, PP (P < 0.01, RR = 0.923), RVEDD/LVEDD (P = 0.006, RR = 3.126) and PASP measured by Echocardiogram (P = 0.002, RR = 1.022), PASP, mPAP, PVR measured by right heart catheter (P = 0.017, RR = 1.001) were predictive. CONCLUSION: IPAH was prone to attack young women with a poor prognosis, the diagnosis of IPAH at an early stage was difficult because of nonspecific symptom, syncope, hepatomegaly and Edema of lower extremity can predict the severity of IPAH, PVR measured by right heart catheterization, PP and PASP were independently predictive factors for IPAH. PMID- 20021923 TI - [Change in ambulatory arterial stiffness index with advancing age and the correlation with 24 hours pulse pressure in healthy individuals]. AB - OBJECTIVE: Ambulatory arterial stiffness index (AASI) has been recently proposed to reflect the dynamic relation between diastolic and systolic blood pressure throughout the whole day. The aim of our study was to investigate the change in AASI with advancing age and the correlation with 24 hours pulse pressure (24 h PP) in healthy individuals. METHODS: 246 healthy subjects [mean age (59.7 +/- 14.6) years, women 38.6%] underwent 24 hours ambulatory blood pressure monitoring (ABPM) in normal life style. The blood pressure recordings, heart rate (HR), mean arterial pressure (MAP) and pulse pressure (PP) were analyzed simultaneously by computer for every 30 minutes during 6:00 am-22:00 pm and every 60 minutes during 22:00 pm-6:00 am. Using all the blood pressure recordings, we plotted diastolic against systolic blood pressure from each individuals and calculated the regression slope. AASI was derived from 1 minus this regression slope. RESULTS: In 246 healthy individuals, AASI increased with age. Among the healthy individuals, the 95th percentile of AASI was 0.56, the upper boundary of the 95% prediction interval of AASI in relation to age were 0.49 at 20 - 39 years, 0.59 at 40 - 59 years, 0.69 at 60 - 79 years, 0.79 at > or = 80 years. The correlation coefficient between AASI and 24 h PP was 0.497 (P < 0.01). AASI linearly increased with age in healthy individuals, whereas the relation between pulse pressure and age was curvilinear. CONCLUSIONS: AASI as a index reflecting blood pressure relationship, manifested the corresponding change with advancing age. The correlation between AASI and traditional index 24 h PP indicated AASI as a new measure of arterial stiffness. PMID- 20021924 TI - [Prognostic value of point of care B-type natriuretic peptide testing and GRACE score in patients with acute coronary syndrome]. AB - OBJECTIVE: To compare the prognostic value of B-type natriuretic peptide (BNP) and GRACE score in patients with acute coronary syndrome. METHODS: A total of 246 patients with chest pain to hospital time < 24 hours were followed up to 30 days. Admission plasma B-type natriuretic peptide was measured by point-of-care. Endpoints included death, reinfarction, recurrent ischemia and new onset of congestive heart failure. The receiver operating characteristic (ROC) curve was used to evaluate prognostic value of BNP and GRACE score. The logistic regression models were used to assess the prognostic contribution of BNP level and GRACE score. RESULTS: The mean age was (67.6 +/- 12.0) years (61.8% males) and ST elevation myocardial infarction (STEMI) was diagnosed in 135 patients (54.9%). During the follow up, 34 endpoints (13.8%) were recorded including 9 deaths (3.7%). The systolic blood pressure [(121 +/- 29) mm Hg vs. (130 +/- 23) mm Hg, P = 0.034; 1 mm Hg = 0.133 kPa] was significantly lower while the heart rate and plasma creatinine were significantly higher in the endpoints group than in non endpoints group. TNI and CRP levels were similar between the two groups. The BNP level at admission (median 883.5 ng/L vs. 216.5 ng/L) and GRACE score (median 164.5 vs. 142.0) were significantly higher in the endpoints group than in non endpoints group (all P < 0.05). The prognostic criteria for BNP level (area under cure, 0.704) was 194.5 ng/L determined by ROC (P = 0.043). For GRACE score, the predictive value for endpoints was 0.742 (P = 0.003) and the cut-off point was 158. In the logistic regression model, BNP concentration (> 194.5 ng/L, OR = 3.174) and GRACE score (> 158, OR = 4.031) were independent predictors of endpoints in patients with ACS. CONCLUSION: Both BNP level at admission and GRACE score were independent predictors for endpoints at 30 days in patients with ACS. PMID- 20021925 TI - [Quantitative coronary angiography and intravascular ultrasound assessments on target lesion and reference vessel in patients with diabetes mellitus]. AB - OBJECTIVE: To evaluate the accuracy of quantitative coronary angiography (QCA) assessment on target lesion and reference vessel in patients with diabetes mellitus with intravascular ultrasound (IVUS) measurements as golden standard. METHODS: QCA and IVUS were performed in 52 diabetes mellitus patients [35 males, mean age (62.3 +/- 7.1) years]. Regression equation was ascertained with the IVUS derived plaque burden as dependent and QCA derived vessel stenosis as independent variable. The measurement results derived from the two modalities on proximal and distal reference vessels were compared. RESULT: The regression equation (constant = 0.8286, P = 0.001) of plaque burden and vessel stenosis derived from two modalities were significantly correlated (r = 0.691, P < 0.001) but QCA overestimated the stenosis severity (57.9% +/- 15.5% vs. 53.5% +/- 12.9%, P < 0.01). Target vessels negative remodeling index in these patient was 0.87 +/- 0.23. QCA significantly underestimated the proximal and distal reference segments vessel diameters [(0.81 +/- 0.24) mm, (0.64 +/- 0.17) mm, all P < 0.05] as compared to IVUS results. CONCLUSION: Due to the significant negative vessel remodeling, QCA overestimated the stenosis severity and underestimated the reference segments vessel diameters in patients with diabetes mellitus. PMID- 20021926 TI - [Eosinophilic coronary arteritis and spontaneous coronary artery dissection: a case report]. PMID- 20021927 TI - [Effect of tirofiban in acute anterior myocardial infarction patients without ST segment resolution after primary percutaneous coronary intervention]. AB - OBJECTIVE: To observe the effect of glycoprotein receptor blockade tirofiban in acute anterior myocardial infarction patients without ST segment resolution after primary percutaneous coronary intervention (PCI). METHODS: From April 2006 to April 2008, 157 acute anterior myocardial infarction patients without ST segment resolution after PCI were randomly allocated to tirofiban (intravenous bolus 10 microg/kg followed by intravenous infusion of 0.15 microgxkg(-1)xmin(-1) for 48 h, n = 80) or equal volume saline (control group, n = 77). Baseline characteristics, PCI features and clinical outcomes during hospitalization, left ventricular ejection fractions (LVEF) and major adverse cardiac events (MACE, including death, re-infarction and target vessel revascularization) at 30 and 180 days after discharge were compared between the two groups. RESULTS: The baseline clinical characteristics were comparable between the two groups. Compared to control group, the MACE rates and re-infarction rates at 30 days (6.3% vs.18.2%, P < 0.05; 1.3% vs.9.1%, P < 0.05, respectively) and 180 days (10.0% vs.23.4%, P < 0.05; 2.5% vs.10.4%, P < 0.05, respectively) were significantly reduced in tirofiban group. LVEF value was significantly higher in tirofiban group at 30 days and 180 days compared with those in control group [(51 +/- 6)% vs. (46 +/- 8)%, P < 0.05; (57 +/- 7)% vs. (50 +/- 9)%, P < 0.05]. Hemorrhagic complications were similar between the two groups. CONCLUSION: Use of tirofiban for acute anterior myocardial infarction patients without ST segment resolution after PCI is safe and can significantly improve 30 and 180 days clinical outcomes after discharge. PMID- 20021928 TI - [Renal dysfunction and survival in hospitalized patients with chronic heart failure: a retrospective analysis]. AB - OBJECTIVE: To analyze the impact of renal dysfunction on survival in hospitalized chronic heart failure (CHF) patients. METHODS: In this retrospective analysis, we collected all clinical data from eligible patients hospitalized in the second hospital of Tianjin Medical University between Jan 1980 and Aug 2007. CHF patients were divided into three groups according to glomerular filtration rate (GFR): A, normal renal function; B, mild renal dysfunction; C, renal dysfunction. Patients in group C were further divided into three subgroups according to hospitalization year: D, 1980.01 - 1989.12; E, 1990.01 - 1999.12; F, 2000.01 - 2007.08. RESULTS: Renal dysfunction was found in 714 patients. Compared with group A (n = 817) and group B (n = 928), patients in group C were older, had worse heart function and major medications included nitrates, diuretics and digitalis. From 1980 to 2007, use of Angiotensin II receptor antagonist, beta blocker, statins significantly increased and the in-hospital mortality significantly decreased in group C patients. Percent of angiotensin converting enzyme inhibitor (ACEI) use was the highest in 1990s. The hospital stay was significantly longer and all cause in-hospital mortality was significantly higher in group C compared to group A and group B (all P < 0.01). After adjustment for other risk factors by multivariate analysis, renal dysfunction was an independent risk factor of in-hospital all cause mortality. Patients faced 16.7% higher risk of all cause in-hospital mortality for every 10 mlxmin(-1) x1.73 m(-2) decrease in GFR. CONCLUSIONS: The incidence of renal dysfunction was high in CHF patients. The hospital stay was longer, in-hospital all-cause mortality was higher in CHF patients with renal dysfunction compared to CHF patients without or with mild renal dysfunction. Renal dysfunction was an independent risk factor for all-cause in-hospital mortality. Increased use of ACEI, ARB, beta-blocker and statins might be responsible for reduced in-hospital mortality in CHF with renal dysfunction patients in recent years. PMID- 20021930 TI - [Novel MYBPC3 mutations in Chinese patients with hypertrophic cardiomyopathy]. AB - OBJECTIVE: To screen the MYBPC3 gene mutations in Han Chinese patients with hypertrophic cardiomyopathy (HCM). METHODS: Sixty-six patients with HCM were enrolled for the study. The exons in the functional regions of MYBPC3 were amplified with PCR and the products were sequenced. RESULTS: Four novel mutations and four common polymorphisms were identified in this patient cohort. A Lys301fs mutation in exon10 was evidenced in a H30, and when he was 47 years old, he had the chest tightness, shortness of breath with septal hypertrophy of 18.7mm; a Asp463stop mutation in exon17 was detected in a H48, he was 24 years old 24-year old when a medical examination showed ventricular septal hypertrophy of 15.4 mm; both Gly523Arg mutation in exon18 and Tyr847His mutation in exon26 were found in a H53 with onset age 36 years old, feeling chest tightness after excise and his ventricular septal hypertrophy was 27 mm that time. MYBPC3 mutations occurred in 4.5% patients in this cohort. These mutations were not found in 100 non-HCM control patients. CONCLUSION: MYBPC3 mutation is presented in a small portion of Han Chinese patients with HCM. PMID- 20021931 TI - [Effects of PPAR-gamma agonist rosiglitazone on MMP-9 and TIMP-1 expression of monocyte-derived macrophages isolated from patients with acute coronary syndrome]. AB - OBJECTIVE: Coronary arterial plaque rupture and secondary thrombosis are the major pathogenesis of acute coronary syndrome (ACS). Metalloprotease (MMPs) secreted by monocyte/macrophage was the main predisposing factor of the plaque rupture and peroxisome proliferator-activated receptor-gamma (PPAR-gamma) is involved in a variety of inflammatory cytokine gene transcriptional regulations. We explored the possible role of PPAR-gamma in the regulation of MMP-9 and TIMP-1 expressed by peripheral monocyte-derived macrophages (MDMs) from patients with ACS. METHODS: Peripheral blood mononuclear cells were isolated from 48 patients with ACS and 28 healthy controls and stimulated by macrophage colony-stimulating factor (0.1 microg/ml for 24 hours) to form MDMs. MDMs were then incubated under various concentrations of rosiglitazone (0, 1, 10, 20 micromol/L) for 48 hours. The concentrations of MMP-9 and TIMP-1 in the supernatant were measured by enzyme linked immunosorbent assay, and the mRNA expression of PPAR-gamma, MMP-9 by RT PCR and nuclear factor-kappaB P65 (NF-kappaB P65) expression by immunohistochemistry. RESULTS: PPAR-gamma mRNA expression was significantly lower while NF-kappaB P65 and MMP-9 expression as well as MMP-9 and TIMP-1 concentrations in supernatant were significantly higher in ACS group than those in control group (all P < 0.05). After rosiglitazone intervention, PPAR-gamma mRNA expression was significantly upregulated in both ACS and control groups in a dose-dependent manner. Both the MMP-9 concentration in the supernatant and MMP-9 mRNA expression were reduced post intervention with rosiglitazone in both groups. The TIMP-1 mRNA expression and concentration in supernatant were not affected by rosiglitazone in both groups. Rosiglitazone induced significant downregulation of NF-kappaB P65 expression in both groups. CONCLUSION: Rosiglitazone intervention may downregulate MMP-9 expression by upregulating PPAR-gamma expression, and by downregulating NF-kappaB expression in MDMs isolated from patients with ACS. PMID- 20021932 TI - [Secretory expression of PR39 following adeno-associated viral-encoding fusion gene transfer induces angiogenesis in hypoxia chick embryo]. AB - OBJECTIVES: To investigate the impact of AAV-encoding NT4-TAT-His-PR39 fusion gene expression on HIF-1alpha level in ECV304 cultured under hypoxic condition (1%O(2)) and on angiogenesis in hypoxic chick embryo. METHODS: PR39 cDNA was connected with NT4, TAT, 6 x His cDNA by molecular biology methods. The recombinant AAV vector was obtained by three plasmid co-transfection in 293 cells. Then ECV304 were respectively infected with AAV-NT4-TAT-His-PR39, 6 x His expression and HIF-1alpha level in ECV304 were detected by immunocytochemistry. The chicken embryos were randomized into the AAV-PR39, EV and PBS groups (n = 10 each) subject to hypoxia (5%O(2), n = 15) or normoxia environments (n = 15), the vessel density of the chicken chorioallantoic membrane (CAM) were measured by Image Pro Plus (IPP) software. RESULTS: The expression of 6 x His protein was detected in AAV-PR39 infected ECV304 cells. HIF-1alpha protein activity was significantly increased in AAV-PR39 infected ECV304 underwent hypoxia compared to PBS and non-infected ECV304 groups (P < 0.05). The vessel density of chicken CAM in hypoxia environment but not in normoxia environment was also significantly higher in AAV-PR39 group than in EV group and PBS group (all P < 0.05). CONCLUSION: AAV-encoding NT4-TAT-His-PR39 fusion gene expression significantly increased HIF-1alpha level in ECV304 exposed to hypoxia and promoted angiogenesis in hypoxic chicken embryo. PMID- 20021933 TI - [Association between heart rate and all-cause death and coronary event in the Chinese cohort: 16 years follow up results]. AB - OBJECTIVE: To evaluate the association between resting heart rate (HR) and all cause death and coronary heart disease (CHD) events in the Chinese cohort. METHODS: Data were obtained from the PRC-USA Cooperative Study on Cardiovascular and Cardiopulmonary Epidemiology. Baseline screen surveys were conducted in 1983 and 1984 from people aged 35 to 59 years living in urban or rural areas of Beijing and Guangzhou. Follow-up visits were performed for end point events of all-cause death and first CHD events every two years till 2005. Resting HR was determined from 5 consecutive intervals between R waves on the 12-lead electrocardiogram. RESULTS: A total of 9856 (4805 males) people were included in the study and the mean follow up duration was 16.2 years. There were 1523 deaths, including 200 CHD events during the follow up period. Mean resting HR was 67.9 beat per minute (bpm) in men and 71.6 bpm in women respectively which had a trend to increase with aging. Cox Proportional Hazards model indicated the relative risk of all-cause death increased constantly with the increase of HR percentile after control of age, fasting glucose, serum cholesterol, serum triglyceride, body mass index, systolic blood pressure and diastolic blood pressure. With HR 60 89 bpm as control group, the relative risk and 95% confidence interval in group HR < 50 bpm, 50 - 59 bpm, 90 - 99 bpm and > or = 100 bpm were 0.76 (0.49 - 1.17), 0.87 (0.75 - 1.02), 1.33 (1.06 - 1.68), 1.48 (1.03 - 2.14) respectively. However there was no significant correlation between HR and CHD events in studied population. CONCLUSION: The risk of total death increased significantly in people with HR > or = 90 bpm suggesting higher resting heart rate might be an independent risk factor for all-cause death in the Chinese population. PMID- 20021934 TI - [A case of giant left atrial thrombus post ablation for atrial tachycardia diagnosed by echocardiography]. PMID- 20021935 TI - [Hypertrophic cardiomyopathy with right aortic arch, right descending aorta, and Kommerell's diverticulum: a case report]. PMID- 20021936 TI - [The successful surgical treatment of a patient with endocarditis after renal transplantation]. PMID- 20021937 TI - [Advances on cardiac stem cell research]. PMID- 20021940 TI - [Gene profiling after knocking-down the expression of NS gene in prostate cancer PC-3 cells]. AB - OBJECTIVE: To screen the genes and possible signal transduction pathways involved in the mechanism of nucleostemin (NS) in the proliferation of prostate cancer. METHODS: Oligonucleotide DNA microarray was used to screen the genome changes after knocking-down expression of NS in PC-3 cells and quantitative real-time PCR was used to further confirm the important differentially expressed genes. RESULTS: 219 differentially expressed genes were found and theses genes were involved in cell cycle, cell proliferation, signal transduction, cell apoptosis and cell differentiation, etc. INK4 family genes (p15, p16, p18) were up regulated and cyclin D1, HDAC1 were down-regulated, the main action points were CDK4/6-cyclin D and pRb-E2F1 complexes. CONCLUSION: NS may promote the progression of prostate cancer by inhibiting the expression of p15, p16, and p18 in PC-3 cells. NS is an important G(1)/S checkpoint regulator and its regulatory activity has been certified at gene level. PMID- 20021941 TI - [Identification of the regions of copy number amplification associated with hepatocellular carcinoma]. AB - OBJECTIVE: To screen and determine the regions of copy number variation (CNV) associated with hepatocellular carcinoma (HCC) using SNP array and fluorescence quantitative PCR. METHODS: The CNV from HCC cell line TJ3ZX-01 was analyzed using GeneChip Human Mapping 500K SNP array. According to the data obtained by SNP array analysis, four candidate amplification regions were verified in 41 primary HCC samples by fluorescence quantitative PCR. RESULTS: Four regions of copy number amplification at 1q21.2, 1q22 approximately 23.1, 7p22.1 and 22q13.1 were detected by SNP array analysis. The four candidate amplicons occurred in 56.1% (23/41) of HCC samples at 1q21.2; 80.5% (33/41) at 1q22 approximately 23.1; 75.6% (31/41) at 7p22.1 and 31.7% (13/41) at 22q13.1 analyzed with sequence tagged site (STS) markers by quantitative PCR. CONCLUSION: In four candidate amplification regions selected by SNP array analysis and detected by fluorescence quantitative PCR, three amplification regions show increased copy number in more than 50.0% HCC tissues. This result indicates that these amplification regions are associated with pathogenesis of hepatocellular carcinoma. PMID- 20021942 TI - [Profile of chromosomal imbalances in 10 cases of primary alveolar rhabdomyosarcoma analyzed by comparative genomic hybridization]. AB - OBJECTIVE: The aim of this study was to characterize the profile of chromosomal imbalances of alveolar rhabdomyosarcoma (ARMS). METHODS: One-step RT-PCR was used to detect the expression of PAX3-FKHR and PAX7-FKHR fusion transcripts in 10 cases of alveolar rhabdomyosarcoma and in an ARMS cell line. Comparative genomic hybridization (CGH) was used to investigate the genomic imbalances in these cases. It was analyzed according to the histological type, pathologic grading, clinical staging, gender and age, respectively. RESULTS: The 10 patients with alveolar rhabdomyosarcoma showed evidence of increased or decreased DNA sequence copy numbers involving one or more regions of the genome. (1) The frequently gained chromosome arms of ARMS were 12q, 2p, 6p, 6q, 10q, 2q, 4q, 15q, 1p, 9q, 14q and 18q (> or = 30.0%), and the frequently lost chromosome arms of ARMS were 3p, 6p, 20q and 21q (> 30.0%). (2) The frequently gained chromosome arm translocation associated with ARMS were 12q, 10q, 2p, 2q, 6p, 6q, 1p, 4q, 8q, 11q, 14q and 15q (> 30.0%). The frequently lost chromosome arms were 3p, 5q, 6p, 1q, 8p, 11p, 20q and 21q (> 30.0%). (3) There were no correlation between chromosome changes and histological type, pathologic grade, clinical stage, gender and age, respectively. CONCLUSION: These observations suggest that: (1) 12q, 2p, 6p, 6q, 10q, 2q, 4q, 15q, 1p, 9q, 14q, 18q gain and 3p, 6p, 20q, 21q loss may correlated with ARMS-related carcinogenesis; (2) 12q gain may be correlated with translocation. PMID- 20021943 TI - [Tumstatin185-191 increases the sensitivity to cisplatin in a cisplatin-resistant human lung adenocarcinoma cell line]. AB - OBJECTIVE: To investigate the effects and related mechanisms of Tumstatin 185-191 as a single agent or in combination with cisplatin on proliferation and apoptosis in a cisplatin-resistant human lung adenocarcinoma cell line A549-DDP cells. METHODS: A549-DDP cells were treated with Tumstatin185-191 and cisplatin at varying concentrations. Cell viability was assessed by a modified 3-(4,5 dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. 50% inhibiting concentration (IC(50)) values of the chemotherapeutic drugs were analyzed by MTT assay. Cell apoptosis was measured by flow cytometry. The activation of Akt and ERK was evaluated by Western blotting. RESULTS: Tumstatin185-191 inhibited the proliferation of A549-DDP cells and its IC(50) value was 80.25 micromol/L. After cotreatment with 20 micromol/L Tum185-191, the IC(50) value of cisplatin in A549 DDP cells reduced from 77.16 micromol/L to 57.97 micromol/L, the reverse index was 1.33, while with 40 micromol/L Tumstatin185-191 the IC(50) was reduced from 77.16 to 26.40 micromol/L and the reverse index was 2.92. The early apoptosis rate was 19.5% +/- 1.1% in the cotreatment group, while 13.3% +/- 1.5% in cisplatin group and 10.2% +/- 2.0% in Tum185-191 group (F = 4.09, P < 0.05). The levels of phospho-Akt (p-Akt) and phospho-ERK (p-ERK) in the A549-DDP cells were remarkably lower after treatment with Tumstatin 185-191. The Tumstatin 185-191 treatment alone or in combination with cisplatin had a similar effect on the protein levels of p-Akt and p-ERK in A549-DDP cells. CONCLUSION: Our data suggest that Tumstatin185-191 may promote apoptosis, downregulate proliferation and partly reverse the drug resistance of A549-DDP cells to cisplatin. The effects induced by Tum185-191 may be mediated through inactivation of the Akt and ERK pathways. PMID- 20021944 TI - [Inhibitory effect of thalidomide on growth of human hepatoma cell line SMMC-7721 cells]. AB - OBJECTIVE: The aim of this study was to investigate the effect of thalidomide on the growth of human hepatoma cell line SMMC-7721 cells in vitro, and to explore the curative possibility of hepatocellular carcinoma with thalidomide. METHODS: SMMC-7721 cells were treated with Thalidomide at different concentrations. The cell growth and proliferation was assessed by MTT assay. DNA ladder, apoptosis rate and changes of cell nuclei were studied by agarose electrophresis, fluorescence microscopy and flow cytometry, respectively. The expression of caspase-3 was analyzed with flow cytometry. The VEGF content of SMMC-7721 cells in culture medium was tested by ELSIA. RESULTS: When the concentration of Thalidomide solution was increased from 3.125 microg/ml to 200 microg/ml, the cell growth was inhibited by from 11.7% to 34.2%. Compared with the control group, the thalidomide solution at a concentration of 25, 50, 100 and 200 microg/ml solution significantly inhibited the proliferation of SMMC-7721 cells (P < 0.05). A ladder pattern of DNA fragments appeared after SMMC-7721 cells exposed to 200 microg/ml thalidomide for 24 h, especially for 48 h. Fluorescence microscopy revealed that the cell nuclei were condensed and fragmented after the cells were exposed to 200 microg/ml thalidomide for 48 h. In cells treated with 200 microg/ml thalidomide for 12, 24, 48 and 72 h, the apoptotic rate was 3.1% +/ 0.5%, 8.4% +/- 1.3%, 19.4% +/- 3.5% and 25.8% +/- 2.1%, respectively, significantly higher than that in the negative control group 1.6% +/- 0.6%. The cells treated with thalidomide at a concentration of 50, 100, 200 microg/ml for 48 h, the apoptotic rate was 8.7% +/- 1.2%, 16.8% +/- 2.5% and 25.4% +/- 4.5%, respectively, increasing in a dose-dependent manner, also significantly than that in the cells of control group 2.1% +/- 0.5%, (all were P < 0.05). The caspase-3 positivity of SMMC-7721 cells treated with thalidomide was increasing along with the increase of treatment time or drug concentration, but not in the control cells. The VEGF content in SMMC-7721 cells was lowering when thalidomide was used in an increasing concentration. CONCLUSION: Under the conditions used in this study, thalidomide can inhibit the proliferation of SMMC-7721 cells in vitro. Induction of apoptosis and inhibition of angiogenesis may be possibly two mechanisms for its anticancer action. PMID- 20021945 TI - [Protective effect of HIF-1alpha-dependent HO-1 overexpression on hypoxic human hepatoma cells in vitro]. AB - OBJECTIVE: To investigate the protective effect of overexpressed heme oxygenase-1 (HO-1) in hypoxia and the correlation between HO-1 overexpressoin and hypoxia inducible factor-1alpha (HIF-1alpha) in human hepatoma HepG2 cells. METHODS: The expressions of HO-1 and HIF-1alpha mRNA as well as the protein were detected by RT-PCR and Western blotting, respectively. MTT assay was used to examine the relative cell survival rate. The total superoxide dismutase (SOD) activity was examined with a SOD kit. RESULTS: Hypoxia induced overexpression of HO-1 gene in HepG2 cells at transcriptional and translational levels. The relative survival rate of HepG2 cells under hypoxia was significantly decreased after the HO-1 protein overexpression was inhibited by ZnPPIX (P < 0.01). The total SOD activity of cells was significantly increased after cells were treated by hypoxia for 16 hours (P < 0.05), while decreased significantly by HO-1 inhibitor ZnPPIX treatment (P < 0.01). HIF-1alpha was upregulated under hypoxia. In addition, the HO-1 overexpression under hypoxia was decreased by HIF-1alpha inhibitor, while the HIF-1alpha expression level under hypoxia was not significantly changed after HO-1 expression was inhibited by ZnPPIX. CONCLUSION: The overexpression of HO-1 in hypoxic HepG2 cells is HIF-1alpha-dependent or at least partly HIF-1alpha dependent. The relative survival rate of hypoxic hepatoma cells was significantly decreased by HO-1 inhibitor treatment. The results of this study may offer new thought and drug target for the therapy of human hepatoma in the future. PMID- 20021946 TI - [A prospective multicenter study of rituximab combined with high-dose chemotherapy and autologous peripheral blood stem cell transplantation for aggressive B-cell lymphoma]. AB - OBJECTIVE: To investigate the feasibility and efficacy of rituximab combined with high-dose chemotherapy supported by autologous peripheral blood stem cell transplantation (ASCT) in patients with aggressive B-cell non-Hodgkin lymphoma (NHL). METHODS: Twenty-eight patients with aggressive B-cell NHL (22 newly diagnosed, 6 relapsed) were enrolled in this study. The high-dose chemotherapy included CHOP regimen (CTX + ADM + VCR + PDN) for the newly diagnosed patients and DICE (DEX + IFO + DDP + VP-16) or EPOCH (VP-16 + PDN + VCR + CTX + ADM) for the relapsed patients. Each patient received infusion of rituximab at a dose of 375 mg/m(2) for four times, on D1 before and on D7 of peripheral blood stem cell mobilization, and on D1 before and D8 after stem cell reinfusion. RESULTS: Complete remission was achieved in all patients after high dose chemotherapy and ASCT. At a median follow-up of 37 months, the estimated overall 4-year survival and progression-free survival rate for all patients were 75.0% and 70.3%, respectively, while both were 72.7% for the previously untreated patients. The therapy was generally well tolerated with few side-effects attributable to rituximab. CONCLUSION: These results suggest that adding rituximab to high-dose chemotherapy with peripheral blood stem cell transplantation is feasible and may be beneficial for patients with aggressive B-cell non-Hodgkin lymphoma. PMID- 20021947 TI - [Study on the mechanism of imatinib-induced resistance in gastrointestinal stromal tumors]. AB - OBJECTIVE: To investigate the mechanism of imatinib mesylate (IM) induced resistance in the patients with gastrointestinal stromal tumors (GISTs) and treated with imatinib. METHODS: Eight patients with GIST treated with IM developed secondary IM resistance. A total of 16 tumor samples (pre-IM therapy) and 11 tumor samples (post-IM treatment) were available. Exon 9, 11, 13, and 17 of c-kit gene as well as exon 12 and exon 18 of PDGFRA gene were sequenced. RESULTS: In addition to the changes of baseline genotype, the IM-induced gene changes were concentrated in the kinase domain of c-kit gene in all 8 patients, 2 of them were located in the exon 13 of c-kit gene presenting with V654A, while 6 in exon 17 involving 816 and 820 to 823 codons. CONCLUSION: The mechanism of imatinib mesylate resistance after initial treatment with this agent in gastrointestinal stromal tumors is a novel mutation development in kinase domain of c-kit. PMID- 20021948 TI - [Local cellular immune response induced by ultrasound-guided tumor bed superantigen injection after percutaneous microwave coagulation therapy for liver cancer]. AB - OBJECTIVE: To investigate the local cellular immune response after injection of superantigen, the highly agglutinative staphylococin (HAS), into the tumor bed after ultrasound-guided percutaneous microwave coagulation therapy (PMCT) in the liver cancer patients. METHODS: Ninety-two patients with pathologically proven primary liver cancer were divided into two groups: 45 in group A were treated by PMCT alone and 47 in the group B by combined with ultrasound-guided percutaneous injection of highly agglutinative staphylococin (HAS). Before and after PMCT and HAS treatment, the patients underwent ultrasound-guided percutaneous biopsy from the tumor bed and the samples were examined by pathology and immunohistochemistry. The infiltration of CD3+, CD4+, CD57+ and CD68+ lymphocytes in treatment zone was compared between the two groups. Moreover, the infiltrating immunocytes were observed by transmission electron microscopy. RESULTS: One week after HAS injection, the densities of CD3+, CD4+, CD57+ and CD68+ cells in the group B were 54.50 +/- 18.44, 38.14 +/- 12.44, 33.38 +/- 10.79 and 45.56 +/- 16.53, respectively. All the above mentioned parameters increased significantly in varying degrees compared with that before PMCT or HAS injection (P < 0.05). Four weeks after HAS injection, the density of CD3+, CD4+, CD57+ and CD68+ cells in the group B were 32.67 +/- 10.42, 23.43 +/- 6.99, 18.63 +/- 7.89 and 30.01 +/- 11.05, respectively, still significantly higher than those before PMCT (P < 0.05). Five weeks after PMCT and HAS injection, the densities of CD3+, CD4+, CD57+ and CD68+ cells in the group B were 54.50 +/- 18.44, 38.14 +/- 12.44, 33.38 +/- 10.79 and 45.56 +/- 16.53, versus 32.03 +/- 8.11, 15.67 +/- 8.32, 15.23 +/- 8.26 and 29.67 +/- 11.98 in the group A, respectively, still with a significant difference between the two groups (P < 0.05). A lot of lysosomes, endoplasmic reticulum and mitochondria in the immune cells after injection of HAS were observed by transmission electron microscopy. CONCLUSION: The local cellular immunity in liver cancer treatment area can be significantly improved by ultrasound-guided injection of highly agglutinative staphylococin after percutaneous microwave coagulation therapy. PMID- 20021949 TI - [Preliminary study of nerve sparing radical hysterectomy in patients with cervical cancer]. AB - OBJECTIVE: To assess the feasibility of nerve sparing radical hysterectomy (NSRH) technique and the impact on the improvement of postoperative bladder function in patients with cervical cancer. METHODS: Forty-two patients with FIGO stage Ib1 approximately Ib2 cervical cancer were selected to receive NSRH (study group, 21 cases) or routine hysterectomy (RH) (control group, 21 cases). Duration of surgery, blood loss and mean length of postoperative stay were compared between the two groups. Immunohistochemical analysis of surgical margins using a general nerve marker (S-100) was performed to compare the nerve damages. RESULTS: The operation time of NSRH group and RH group was (248 +/- 24) min and (227 +/- 27) min, respectively, with a significant difference between the two groups (P < 0.01). No significant difference in blood loss was found between the NSRH and RH group [(459 +/- 143) ml vs. (454 +/- 121) ml, P > 0.05]. However, the median urinary catheterization time was 7 days in NSRH group versus 16 days in the RH group, with a statistically significant difference between the two groups (P < 0.01). The rate of patients who had postoperative residual urine volume in bladder (PVR) < or =100 ml was 66.7% in the NSRH group versus 19.0% in the RH group, with a significant difference between the two groups (P < 0.01). No severe perioperative complications occurred in both groups. After a follow-up of 11 to 16 months (median: 14 months), no recurrence was detected in the two groups. Immunohistochemistry with S-100 staining revealed only small nerve fibers in the surgical margins of the NSRH group, but full with large nerve bundles in that of the RH group. There was a significant difference between two group (P < 0.01). CONCLUSION: The results of our preliminary study indicate that nerve sparing radical hysterectomy (NSRH) for the patients with FIGO stage Ib1 approximately Ib2 cervical cancer is safe and feasible, and can well preserve the pelvic autonomic nerves and improve the recovery of bladder voiding function. PMID- 20021950 TI - [Prognostic factors and outcome in patients with intrahepatic recurrence after hepatectomy for hepatocellular carcinoma]. AB - OBJECTIVE: To investigate the prognostic factors and treatment choice for intrahepatic recurrence after hepatectomy in patients with hepatocellular carcinoma (HCC). METHODS: Clinicopathological data of 184 HCC patients with intrahepatic recurrence after hepatectomy were collected. The influences of twenty one clinicopathological factors and treatment modalities on the survival after recurrence were retrospectively analyzed. RESULTS: Univariate analysis showed that preoperative serum alpha-fetoprotein (AFP) >100 ng/ml, microscopic venous invasion, patients classified as Child-Pugh class B or C at diagnosis of recurrence, multiple recurrence foci and early recurrence (< or =12 months) were poor prognostic factors. Cox multivariate analysis showed that Child-Pugh class at diagnosis of recurrence, number of recurrent foci and time to recurrence were independent risk factors for survival in patients with recurrence. Median survival after recurrence was 34 months, 23 months, 15 months and 9 months, respectively, in patients treated by repeated hepatectomy, local ablation therapy, transcatheter arterial chemoembolization (TACE) or non-treatment in 69 patients with solitary recurrence. There were statistically significant differences among these four groups (P < 0.05). CONCLUSION: classification of Child-Pugh class A at the first time of diagnosis, solitary recurrence, late recurrence (> 12 months), and intrahepatic recurrence occurred after repeated hepatectomy or local ablation therapy are better prognostic factors in patients with HCC recurrence. PMID- 20021951 TI - [Influence of active breathing control on the dose distribution in the target of forward whole-breast intensity-modulated radiotherapy after breast conserving surgery]. AB - OBJECTIVE: To explore the influence of intrafraction and interfraction target displacement on the dose distribution in the target of forward whole-breast intensity-modulated radiotherapy (IMRT) assisted by active breathing control (ABC). METHODS: Each of the selected patient who had received breast conserving surgery was immobilized and received the primary CT simulation assisted by ABC device to get five sets of CT images in three different breathing status, including free breathing (FB) (one set), moderate deep inspiration breathing hold (mDIBH)(two sets) and deep expiration breathing hold (DEBH) (2 sets). After 10 to 15 fractions of radiation, the repeated CT simulation was completed and the same five sets of CT images were obtained at FB, mDIBH, and DEBH, respectively. In the Pinnacle3 treatment planning system, the forward IMRT planning was completed on the first set of mDIBH CT images from the primary CT simulation, and the planning was separately copied by the special system order to the second set of CT images from the primary CT simulation and to the first set of CT images from the repeated CT simulation, keeping the primary angle, direction, size and shape of the MLC field and prescribed dose un-changed. the volumes covered by selected high dose area in the selected segment were compared. RESULTS: In the planning based on the first set of mDIBH CT images from the primary CT simulation, the volume irradiated by equal and more than 103% of prescribed dose in the segment was (1.16 +/- 0.39) cm3, and the volumes were (3.88 +/- 1.07) cm3 and (51.66 +/- 8.68) cm3 in the plannings copied from the first set of mDIBH CT images from the primary CT simulation respectively to the second set of CT images from the primary CT simulation and first set of CT images from the repeat CT simulation, the difference of the volume in the two plannings based on the two set mDIBH CT image from the primary CT simulation was not statistically significant (t = 1.672, P = 0.103). The difference of the volume in the two plannings based on the two sets of mDIBH CT images respectively from the primary and repeat CT simulations had a significant difference (t = -5.728, P < 0.01). CONCLUSION: If the same threshold of mDIBH is maintained during IMRT after breast conserving surgery, the influence of the intrafraction target displacement on the dose distribution is not significant. However, if set-up error is not adjusted, the interfraction change of position of the segment given to cover the high dose area in the IMRT planning will be significant, resulting in a significant change of dose distribution in the breast. PMID- 20021952 TI - [Application of standardized uptake value for FDG PET-CT in predicting radiation pneumonitis]. AB - OBJECTIVE: To investigate the correlation of radiation pneumonitis (RP) with standardized uptake value (SUV) for fluorodeoxyglucose (FDG) positron emission tomography and computed tomography (PET-CT) in lung cancer patients treated with radiation therapy. METHODS: Fourty patients with unresectable non-small cell lung cancer (NSCLC) received FDG PET-CT before and after radiotherapy. The average SUV of the lung tissue irradiated with a dose of < or = 5 Gy, 5.1 approximately 15 Gy, 15.1 approximately 35 Gy, 35.1 approximately 60 Gy, >60 Gy were measured. The correlation between SUV and RP was analyzed by comparing the SUV in the patients with RP and without. The SUV ratio of the irradiated lung tissue to that of the non-irradiated lung tissue (L/B) was also calculated. RESULTS: Of the 40 patients, 8 developed RP, including 6 cases of grade 2 and 2 cases of grade 3. The SUV of irradiated lung tissues with a dose of 35.1 approximately 60 Gy was significantly correlated with RP. When SUV > or =1, the RP incidence rate was 41.7% versus 20.0% in the whole group, with a statistically significant difference. (chi2 = 3.96, P < 0.05). The sensitivity and specificity of SUV in predicting RP was 62.5% and 78.1%, respectively. When the value of L/B > or = 2.5, the RP incidence rate was 40.7% in this group versus 20.0% in the whole group, with a statistical significance (chi(2) = 4.92, P < 0.05). If taking L/B > or = 2.5 as a threshold value, the sensitivity and specificity in predicting RP was 72.7% and 90.9%, respectively. No statistically significant difference was found in predicting radiation pneumonitis between SUV > or =1 and L/B > or = 2.5 (chi2 = 0.002, P > 0.05). CONCLUSION: The standardized uptake value (SUV) and the SUV ratio of the irradiated lung tissue to that of the non-irradiated lung tissue (L/B) for FDG PET-CT are positively correlated with radiation pneumonitis, and clinicians may use it to predict the occurrence of radiation pneumonitis. PMID- 20021953 TI - [Clinical value of "Kou mode of hepatic hilar anastomosis" in resection of type III or IV hepatic hilar cholangiocarcinoma]. AB - OBJECTIVE: To evaluate the surgical technique of "Kou mode of hepatic hilar anastomosis" in the treatment for type III or IV hilar cholangiocarcinoma. METHODS: The clinical data of 89 patients with type III or IV hilar cholangiocarcinoma surgically treated in our department between Jan. 1990 and Jan. 2008 were retrospectively analyzed. Since January 2000, "Kou mode of hepatic hilar anastomosis" was performed for some patients with advanced hilar cholangiocarcinoma. The patients were divided into two groups: group A treated between 1990 and 1999, group B between 2000 and 2008. The rate of resection, therapeutic efficacy and complications in these two groups were compared, respectively. RESULTS: Of the 37 cases with hilar cholangiocarcinoma in group A, 4 were surgically treated (10.8%), with 1 (2.7%) radical resection and 3 (8.1%) palliative resection. Among the 52 cases with hilar cholangiocarcinoma in the group B, 35 (67.3%) received surgical resection, of them 15 (28.8%) underwent radical resection and 20 (38.5%) had palliative resection. Twenty-eight of these 35 cases underwent the "Kou mode of hepatic hilar anastomosis". The resection rate of advanced hilar cholangiocarcinoma in the group B was significantly higher than that in group A (P < 0.05). The complications in the 89 cases included ascites (3 cases), hemobilia (1 case), heart failure (1 case), and wound infection (2 cases). All the patients who were treated with the "Kou mode of hepatic hilar anastomosis" developed bile leakage to a varying degree and recovered after drainage and symptomatic treatment. CONCLUSION: The resection rate of type III or IV advanced hilar cholangiocarcinoma can be remarkably improved by using a novel alternative surgical technique called "Kou mode of hepatic hilar anastomosis". However, the long-term outcome still needs to be determined by close follow-up and further observation. PMID- 20021954 TI - [Analysis of the therapeutic effect of esophagectomy with extended 2-field lymph node dissection for esophageal carcinoma]. AB - OBJECTIVE: To summarize the surgical effect and clinical application value of esophagectomy with extended 2-field lymph node dissection for patients with esophageal carcinoma. METHODS: From June 1987 to December 2008, 1690 patients with esophageal cancer underwent esophagectomy with extended 2-field (thoracic and abdominal) dissection of lymph nodes. Patients with the middle and lower thoracic esophageal cancer underwent Ivor-Lewis esophagectomy, and patients with upper thoracic esophageal cancer underwent Akiyama esophagectomy. 2-field (thoracic and abdominal) lymph node metastases information and the 1, 3, 5, 10 year survival rates were analyzed retrospectively. RESULTS: Lymph node metastases were found in 713 patients. The lymph node metastases rate was 42.2% (713/1690).Thoracic lymph node metastasis rate was 39.3% (665/1690), among which in the right pleural apical para-tracheal triangle was 20.7% (349/1690), in the posterior upper mediastinum was 26.3% (444/1690), in the lower mediastinum was 18.2% (307/1690). Abdominal lymph node metastasis rate was 20.1% (339/1690). THE Postoperative complication rate was 16.4% (278/1690), among which the pulmonary complication rate ranking the first, was 43.6% (136/312). The operative mortality rate was 0.2%. The 1-year, 3-year, 5-year and 10-year survival rates were 88.2% (1388/1574), 63.5% (868/1367), 54.8% (705/1287) and 30.8% (232/754), respectively. The 5-year survival rate in patients without lymph node metastasis was 76.2% (448/588), but that in patients with lymph node metastases was 36.8% (257/669). CONCLUSION: The results of this study demonstrated that Ivor-Lewis and Akiyama esophagectomy with two-field lymph node dissection exposes the operation fields clearly and make radical lymphadenectomy thoroughly, especially the lymph nodes in the posterior upper mediastinum around the recurrent laryngeal nerve and in the right pleural apical para-tracheal triangle. It is essential that patients with esophageal carcinoma with lymph node metastases should undergo esophagectomy with extended 2-field dissection of lymph nodes. This can elevate the postoperative 5-year survival rate remarkably. PMID- 20021955 TI - [Surgical treatment for bronchioloalveolar carcinoma with ipsilateral intrapulmonary metastatic nodules]. AB - OBJECTIVE: The staging and treatment of bronchioloalveolar carcinoma (BAC) with pulmonary metastasis are still controversial. This study aimed at evaluating the current staging of BAC with ipsilateral intrapulmonary metastatic nodules and the therapeutic effectiveness of surgical resection. METHODS: The clinicopathological data of 729 completely and surgically resected patients with non-small cell lung cancer (NSCLC) from December 1999 to December 2006 were retrospectively reviewed. Prognostic factors affecting the overall survival were analyzed by the Kaplan Meier method and compared by the log rank test. RESULTS: Among 67 NSCLC patients with ipsilateral intrapulmonary metastatic nodules, 54 had multiple nodules in the lobe with primary lesion (T4, PM1) and 13 had additional nodules in the other ipsilateral lobes (M1, PM2). This series consisted of 40 males and 27 females, with a median age of 60.0 years. Of those, 28 had the lesions containing pure or some bronchioloalveolar carcinoma component, while the other 39 had a NSCLC lesions containing non-bronchioloalveolar carcinoma components. The median overall survival time of this series was 24.0 months. Prognostic study demonstrated that bronchioloalveolar carcinoma histology and mediastinal lymph node metastasis had significant adverse impact on the overall survival. The median survival time of the patients with bronchioloalveolar carcinoma was 58.0 months versus 27.0 months in patients with other subtypes of NSCLC (P < 0.01). The median survival times were 39.0 months for the patients with N0 or N1 versus 14.0 months for patients with N2, with a significant difference between the two groups (P < 0.01). There was no significant difference in the survival time between the patients with PM1 (36 months) and those with PM2 (24 months) (P > 0.05). CONCLUSION: Surgical resection is effective for NSCLC patients with ipsilateral intra-pulmonary metastasis, especially for those with bronchioloalveolar carcinoma components. Our results suggest that the current TNM classification system may be inappropriate for the NSCLC patients with ipsilateral intrapulmonary metastatic nodules, and may need a modification. PMID- 20021956 TI - [To explore a route to liver cancer research with Chinese features]. PMID- 20021957 TI - [The biological features and their roles of cancer stem cells in invasion and neovascularization of cancer]. PMID- 20021958 TI - [Recent advances in studies on vascular progenitor cells]. PMID- 20021959 TI - [An update: stem cell and cancer stem cells]. PMID- 20021960 TI - [Relationship between primary ocular adnexal mucosa-associated lymphoid tissue lymphoma and eye infection]. AB - OBJECTIVE: To study the role of pathogenic microorganisms commonly associated with chronic eye disease, including Chlamydia psittaci, Chlamydia trachomatis, Chlamydia pneumoniae, herpes simplex virus (HSV) type 1 and type 2, and adenovirus type 8 and type 19, in the development of primary ocular adnexal mucosa-associated lymphoid tissue (MALT) lymphoma in Chinese patients. METHODS: Sixty-eight archival cases of primary ocular adnexal lymphoproliferative lesions, including 38 cases of MALT lymphoma, 3 cases of non-MALT lymphoma and 27 cases of chronic inflammation, were enrolled into the study. DNA was extracted from the paraffin-embedded tissue samples. The presence of DNA of C. psittaci, C. trachomatis, C. pneumoniae, HSV type 1, HSV type 2, adenovirus type 8 and adenovirus type 19 were analyzed by multiplex touchdown enzyme time-release polymerase chain reaction (TETR-PCR). RESULTS: All of the specimens yielded PCR products of over 100 base pairs and were thus suitable for TETR-PCR screening of infectious agents. The prevalence of DNA of C. psittaci, C. trachomatis and adenovirus type 19 were 0 in MALT lymphoma, non-MALT lymphoma and chronic inflammation. There were 2 cases positive for C. pneumoniae DNA, amongst the 38 cases of MALT lymphoma studied (5.3%, 2/38). HSV type 1, HSV type 2 and adenovirus type 8 DNA was found in each of the 3 patients with chronic inflammation. CONCLUSION: The study indicates that C. psittaci, C. trachomatis, C. pneumoniae, HSV type 1, HSV type 2, adenovirus type 8 and adenovirus type 19 probably play little role in the pathogenesis of ocular adnexal MALT lymphoma in Chinese patients. PMID- 20021961 TI - [Relationship between epithelial-mesenchymal transition and basal cell-like phenotype in breast cancer]. AB - OBJECTIVES: To evaluate the relationship between epithelial-mesenchymal transition and basal cell-like phenotype breast cancer (BLBC). METHODS: Three hundred and eighty two cases of breast cancers including basal cell-like, luminal A, luminal B and Her-2 subtypes were collected from 458 cases of invasive breast cancers based on their immunophenotypes. They were then stained immunohistochemically with FOXC-2, vimentin, Syndecan-1 and E-cadherin. The relationship of these markers with the basal cell-like phenotype of breast cancer was studied. RESULTS: Of the 41 BLBC cases, FOXC-2, vimentin and Syndecan-1 were positive in 14 cases (34.1%), 18 cases (43.9%) and 36 cases (87.7%) respectively; E-cadherin expression was reduced in 26 cases (63.4%). The positive rates of FOXC 2 and vimentin were higher in BLBC than in other subtypes of breast cancer (P < 0.01). The expression of E-cadherin was the lowest among the 4 subtypes of breast cancers (P < 0.01). Syndecan-1 was positive in the stromal cells adjacent to cancer cells in 17 cases (41.5%) BLBC and the expression was higher than that in other subtypes (P = 0.007). There existed a correlation between FOXC-2 and vimentin expression in BLBC (r = 0.607, P < 0.01). The rates of positive lymph nodes in FOXC-2 and vimentin positive BLBC cases were 71.4% and 66.7% respectively, and both were higher than those of FOXC-2 and vimentin negative BLBC cases (P = 0.002 and P = 0.001). CONCLUSION: Epithelial-mesenchymal transition is probably related to the basal cell-like phenotype of breast cancers, and this may be one of the reasons accounting for the different biological behavior of BLBC from other subtypes of breast cancer. PMID- 20021962 TI - [Expression of CD147, matrix metalloproteinases and transforming growth factor beta1 in breast cancer]. AB - OBJECTIVE: To investigate the expression of CD147, matrix metalloproteinase (MMP) 2, MMP-9, Transforming growth factor (TGFbeta1) and TGFbetaRI proteins and their relationships to breast cancer. METHODS: The expression of CD147, MMP-2, MMP-9, TGFbeta1 and TGFbetaRI proteins was examined on tissue chips containing 160 cases of breast carcinomas by S-P immunohistochemical method. RESULTS: The positive rates of CD147, MMP-2, MMP-9, TGFbeta1 and TGFbetaRI proteins were 87.5% (140/160), 96.9% (155/160), 95.0% (152/160), 73.7% (118/160) and 60.6% (97/160), respectively. The expression of CD147 was positively correlated with axillary lymph node metastasis, TNM staging and HER2 over expression (P < 0.01, P < 0.05 and P < 0.01, respectively), and inversely correlated with PR expression (P < 0.05). The patients' relapse-free survival was shorter in TGFbeta1-positive group than in TGFbeta1 negative group (P < 0.05). Both the expression of MMP-2 and MMP 9 were positively correlated with CD147 expression; and both the expression of TGFbeta1 and TGFbetaRI were positively correlated with CD147 expression (P < 0.01 and P < 0.05, respectively). CONCLUSION: The expression of CD147 is considered closely correlated with tumor invasion, metastasis and prognosis in breast cancer, and has also a close correlation with MMP-2, MMP-9, TGFbeta1 and TGFbetaRI expression. PMID- 20021963 TI - [Effects of protein kinase CK2alpha on the proliferation and invasion of human nasopharyngeal carcinoma]. AB - OBJECTIVE: To investigate the effect of protein kinase CK2alpha expression on the proliferation and invasion of human nasopharyngeal carcinoma and related mechanism. METHODS: RNA interference (RNAi) was used to down-regulate the protein kinase CK2alpha expression in 5-8F cells. The biological behavior of treated cells were analyzed by MTT and in-vitro invasion assays. Cell proliferation cycle was examined by flow cytometry and the phosphorylation status of Akt protein was examined by Western blotting analysis. RESULTS: CK2alpha protein was successfully silenced by siRNA. CK2alpha knockdown significantly decreased the proliferative and invasive abilities of 5-8F cells. Flow cytometry analysis showed that the percentage of cells increased in G(0)/G(1) phase but decreased in S phase. Moreover, the expression of phosphorylated Akt was down-regulated by CK2alpha silencing. CONCLUSION: Protein kinase CK2alpha plays an important role in the proliferation and invasion of nasopharyngeal carcinoma, and may provide a potential therapeutic target against nasopharyngeal cancer. PMID- 20021964 TI - [High glucose and tea polyphenols on expression of matrix metalloproteinase-2 and extracellular matrix metalloproteinase inducer in human umbilical vein endothelial cells]. AB - OBJECTIVE: To investigate the effects of high glucose on expression of matrix metalloproteinase-2 (MMP-2) and extracellular matrix metalloproteinase inducer (EMMPRIN) in human umbilical vein endothelial cells (HUVECs). METHODS: HUVECs were cultured in DMEM media containing high glucose with or without polyphenols for 24 hours respectively. The activity of MMP-2 in the supernatant was detected by zymography. The expression of MMP-2 mRNA and protein in HUVECs were detected by RT-PCR and Western blot respectively. The expression of EMMPRIN mRNA and protein in the cells were determined by RT-PCR as well as immunocytochemistry and Western blot respectively. RESULTS: The expression of MMP-2 and EMMPRIN mRNA were suppressed by a high concentration of glucose. Both the MMP-2 activity and protein level of EMMPRIN expression were also significantly decreased. Polyphenols abolished all the above changes of HUVECs induced by a high glucose level (P < 0.05). CONCLUSIONS: An acute high glucose stimulation down-regulates the activity of MMP-2, the expressions of MMP-2 and EMMPRIN at RNA and protein levels in the endothelial cells, which may play an important roles in diabetic vascular complications in the early phase. Polyphenols treatment can diminish the detrimental effects of high glucose on HUVECs. PMID- 20021965 TI - [Expression and function of VAP-33 in murine dendritic cell sarcoma]. AB - OBJECTIVE: To elucidate the expression and function of VAP-33 gene in dendritic cell sarcoma (DCS) cell line. METHODS: The expression of VAP-33 in DCS cells was investigated by mass spectrum with immunoprecipitation membrane protein. DCS cells were treated with antigens in different dosages (150, 850, and 1500 microl) for 24, 48 and 72 h respectively. Cell morphology and phagocytosis activity of DCS cells were measured. Indirect immunofluorescence, confocal microscopy and Western blotting were used to study the distribution and expression changes of VAP-33. Moreover, DCS cells were treated with 0.5 mol/L insulin for 20 min first and followed by Western blotting to detect changes of VAP-33 and glucose transfer protein 4 (GLUT-4) in the total cellular protein, cytoplasmic protein and membrane protein. Confocal microscopy was used to document the expression and distribution changes of VAP-33 and GLUT-4 in DCS cells. RESULTS: VAP-33 expression was obtained at the cell membrane and in the cytoplasm of DCS cells. Upon antigen stimulation, DCS cells showed more active phagocytosis and morphologically became more elongated with branched protrusions. The expression of VAP-33 was decreased by the antigen stimulation. Upon the insulin stimulation, the expression of VAP-33 and GLUT-4 were increased and co-localized. CONCLUSIONS: VAP-33 expression in DCS originated from the dendritic cells (DC) seemed relating to the vesicle transportation during antigen processing in DC. Additionally, VAP 33 and GLUT-4 also take part in the glucose transportation in the cells. PMID- 20021966 TI - [Diagnosis of hematolymphoid malignancy by using effusion fluid cytology specimens: a study of 33 cases]. AB - OBJECTIVE: To study the diagnostic accuracy of hematolymphoid malignancy by using effusion fluid cytology specimens and to evaluate the values of immunocytochemistry for this assay. METHODS: The cytospin preparations/smears and cell block sections of effusion cytology specimens from 33 cases of hematolymphoid malignancy were retrospectively reviewed. Immunocytochemical study was performed. In selected cases, in-situ hybridization for Epstein-Barr virus encoded RNA and immunoglobulin and T-cell receptor gene rearrangement study were carried out as indicated. RESULTS: There were 33 cases of hematolymphoid malignancy, including 12 cases of T-lymphoblastic leukemia/lymphoma, 16 cases of mature B cell neoplasm (including 9 cases of diffuse large B-cell lymphoma, 2 cases of Burkitt lymphoma, 2 cases of plasmacytoma/multiple myeloma, 2 cases of B small lymphocytic leukemia/lymphoma and 1 case of mantle cell lymphoma), 3 cases of mature T or NK-cell neoplasm (including 1 case of extranodal nasal NK/T-cell lymphoma, 1 case of angioimmunoblastic T-cell lymphoma and 1 case of T-cell prolymphocytic leukemia), 1 case of myeloid sarcoma and 1 case of mast cell sarcoma. Amongst the 33 cases studied, 16 represented disease relapses, including 8 cases of diffuse large B-cell lymphoma, 2 cases of plasmacytoma/multiple myeloma, 2 cases of B-small lymphocytic leukemia/lymphoma, 1 case of T lymphoblastic leukemia/lymphoma, 1 case of angioimmunoblastic T-cell lymphoma, 1 case of mantle cell lymphoma and 1 case of mast cell sarcoma. The remaining 17 cases showed serous effusion as the primary manifestation, with the diagnosis primarily made upon cytologic examination. The cytologic findings seen in all the 33 cases studied were in agreement with the corresponding histologic diagnosis. CONCLUSIONS: Diagnosis of hematolymphoid malignancy by effusion fluid cytology specimens is possible, especially when coupled with the clinical history, immunophenotype, in-situ hybridization and gene rearrangement study findings. This is especially so for cases with disease relapses. PMID- 20021967 TI - [Evaluation of immunohistochemistry staining and cytologic diagnosis by using cell block sections prepared with effusion fluid cytology specimens]. AB - OBJECTIVE: To study the values of immunohistochemistry staining and cytological diagnosis by using cell block sections prepared with the effusion fluid cytology specimens. METHODS: Ninety-nine effusion cytology specimens with the diagnoses of reactive mesothelial hyperplasia, atypical cells and metastatic carcinoma were enrolled into the study. The cytospin preparations/smears, cell block sections and immunohistochemical study were performed and correlated with the clinical findings and follow-up data. RESULTS: Amongst the 99 cases studied, the percentage with positive diagnosis using cytospin preparations/smears was 68.7% (68/99). The percentages with negative and equivocal diagnoses were 16.2% (16/99) and 15.1% (15/99), respectively. As for cell block sections, the percentages were 71.7% (71/99), 16.2% (16/99) and 12.1% (12/99), respectively. On the other hands, the percentages became 76.8% (76/99), 20.2% (20/99) and 3.0% (3/99), respectively, when coupled with immunohistochemical findings. The overall percentages of positive, negative and equivocal diagnoses were 77.8% (77/99), 17.2% (17/99) and 5.0% (5/99), respectively, upon clinicopathologic correlation. The difference between cytospin preparations/smears and cell block sections was not statistically significant (P > 0.05). When coupled with immunohistochemical findings or clinicopathologic correlation, the difference in rates of equivocal diagnosis however carried statistical significance (P < 0.05). The false-negative rate of immunohistochemical study applied on cell block sections was 1.0% (1/99). CONCLUSIONS: Immunohistochemistry, when applied on cell block sections, is useful in delineation of the primary origins of the tumor cells in effusion fluid cytology specimens. Combination of morphologic examination, immunohistochemical findings and clinicopathologic correlation can further improve the rate of positive diagnosis. PMID- 20021968 TI - [Relationship between mitochondrial gene changes and cardiomyocytic injury in experimental ischemia/reperfusion in rats]. PMID- 20021969 TI - [Immunophenotype analysis on neoplastic cells in bone marrow and peripheral blood of angioimmunoblastic T-cell lymphoma]. PMID- 20021970 TI - [Primary leiomyosarcoma upper end of tibia: report of a case]. PMID- 20021971 TI - [Thymoma associated with an lipofibroadenoma: report of a case]. PMID- 20021972 TI - [Uterine papillary serous carcinoma: report of a case]. PMID- 20021973 TI - [Paratesticular desmoplastic small round cell tumor: report of a case]. PMID- 20021974 TI - [Superficial angiomyxoma: report of a case]. PMID- 20021975 TI - [Clear cell meningioma: report of a case]. PMID- 20021976 TI - [Advance in biomarkers of lung cancer in diagnosis and targeted therapy]. PMID- 20021977 TI - [Use of tumor proliferation marker ki-67 and PCNA in surgical pathology]. PMID- 20021978 TI - Effect of progesterone and pregnancy on the development of varicose veins. AB - The aim of this study was to investigate the contribution of progesterone in the development of primary varicose veins on lower limbs during pregnancy. In 50 primiparae with varicose veins, serum progesterone level was quantitatively determined in the 14th week of pregnancy and results were compared with those obtained in a control group of 25 primiparae without visible varicose veins. The mean serum progesterone concentration recorded in pregnant women with dilated veins (159.9+/-15.8 nmol/L) was significantly higher as compared with the control group (159.9+/-15.8 nmol/L vs. 40.4+/-1.6 nmol/L; P<0.0001). These findings supported the role of hormonal factor in the development of varicose veins in women. PMID- 20021979 TI - Microbiological findings in prepubertal girls with vulvovaginitis. AB - The aim of the study was to define the most common causes, symptoms and clinical features of vulvovaginitis in prepubertal girls, and to evaluate treatment success depending on the causative agent involved. The study included 115 girls aged 2-8 (mean 4.8) years, presenting with vulvovaginitis to the Outpatient Clinic for Pediatric and Adolescent Gynecology, Zagreb Children's Hospital, between September 2006 and July 2007. Medical history data were obtained from parents. Vaginal samples were collected for microbiological culture by using cotton-tipped swabs moistened with saline. All samples were referred to microbiology laboratory, where standard microbiological diagnostic procedures were performed. Selective and non-selective media were used. Of 115 study patients, 43 (37.4%) had received antibiotic therapy more than one month prior to their visit to the Clinic, mainly for upper respiratory tract infection. The most common presenting symptom was increased vaginal discharge usually noticed on the pants or diaper, found in 26 of 115 (22.6%) patients, followed by vulvar redness in 16 (13.9%), burning in seven (6.1%), itching in the vulvovaginal area in seven (6.1%), soreness in six (5.2%), odor in three (2.6%) patients, and two or more of these symptoms in another 50 (43.5%) patients. Fifty-nine of 115 children had normal clinical finding on gynecologic examination. Among the remaining 56 children, the most common finding was erythema observed in 19, vaginal discharge in ten, and a combination of discharge and erythema in 13 patients. Of 115 study patients, causative agents were isolated from vaginal culture in 38 (33%) cases. Of these, 21 grew group A beta hemolytic streptococcus, five patients Haemophilus influenzae, three Escherichia coli, two Enterococcus spp., and one each Staphylococcus aureus, Proteus mirabilis, and Streptococcus pneumoniae. Antibiotic therapy was administered in 31 of these 38 patients, except for those cases where intestinal bacteria and Staphylococcus aureus were isolated and topical therapy and hygienic measures were applied alone. Accordingly, vulvovaginitis in girls was most commonly caused by pathogens originating from the patient upper respiratory tract, accompanied by the symptoms of redness and vaginal discharge. In these cases, antibiotic treatment was administered. In the majority of prepubertal girls with either vulvitis or normal genital finding, simple measures to improve hygiene will lead to resolution of all symptoms. PMID- 20021980 TI - Treatment of stable vitiligo by ReCell system. AB - The aim of our study was to analyze the results obtained with the ReCell system for surgical treatment of stable vitiligo. At Department of Plastic and Reconstructive surgery, University of Tor Vergata in Rome, we treated 15 patients with stable vitiligo during a 2-year period. The stability of vitiligo lesions varied between 1 and 4 years. The mean stability was 2.25. The disease activity according to Vitiligo Disease Activity Score (VIDA) was 0 in all cases, which means that all patients had a stability of greater than or equal to 1 year. Three (20%) patients had vitiligo vulgaris, seven (46.6%) segmental vitiligo and five (33.3%) focal vitiligo. Repigmentation was assessed using the Vitiligo Area Scoring Index (VASI). The extent of pigmentation was scored as excellent, good, fair, and poor depending on the percentage of repigmentation in the previously depigmented site. The color of the repigmented area was compared with the adjacent normally pigmented area: excellent color match was present in ten (66.6%) and good in five (33.3%) cases. There was no fair or poor outcome. Repigmentation greater than 75% was recorded in 12 (80%) and 25% to 50% repigmentation in three (20%) of 15 patients treated. ReCell is a feasible, simple and safe technique. The method that uses noncultured autologous epidermal suspension is simpler, less expensive, less time consuming, and does not require sophisticated laboratory facilities as compared with the methods employing cultured melanocytes. PMID- 20021981 TI - History of venereology in Croatia. AB - The term sexually transmitted infections has a wider meaning than the older venereal diseases. It is known that gonorrhea was present even in ancient times, whereas syphilis appeared in Europe at the end of the 15(th) century. Soon cases were observed along the coast in Zadar, Dubrovnik and Istria, and were treated by surgeons. The relatively late establishment of medical schools, along with wars and different governments, were some of the reasons why dermatovenereology as a specialty developed later in Croatia than in other European countries. Nevertheless, during the 19(th) century, many Croatian physicians and dermatovenereologists of the past century were interested in venereology. Among the most important personalities that worked at departments and dispensaries for venereal diseases were Cambieri, Kogoj, Bozic, Nonveiller, Orhel, and many others. They introduced new diagnostic technologies and treatments, and wrote about these diseases. Significant progress was achieved in larger towns where medical schools had been opened. Croatian venereologists also organized several international congresses on sexually transmitted diseases and became members of the respective international societies. Nowadays, the interest of dermatovenereologists has been shifting towards chlamydial infections, herpes genitalis, human papillomavirus genital infection and human immunodeficiency virus infection, and their presence among homosexuals. PMID- 20021982 TI - Iron status in diffuse telogen hair loss among women. AB - The relationship between iron body status and different types of hair loss has been investigated in a number of studies, however, with relatively discrepant findings. Therefore we conducted an analytical case-control study to assess whether diffuse telogen hair loss in women of childbearing age (15 to 45 years old) is associated with iron deficiency. Using the analytical case-control methodology, we studied 30 consecutive women with documented diffuse telogen hair loss in comparison with 30 women without hair loss. Study subjects had no history of nutritional supplement intake or chronic underlying diseases, and had normal thyroid function and inflammatory profiles. Biochemical investigations were performed in all study women. The mean ferritin level and trasferrin saturation was statistically significantly lower in patients with diffuse telogen hair loss than in subjects without hair loss (16.3+/-12.6 vs. 60.3+/-50.1, ng/mL; P<0.0001 and 20.3+/-9.7 vs. 28.3+/-11.8 percent; P=0.006, respectively). Also, total iron binding capacity was significantly higher in patients than in control group (367.8+/-58.2 vs. 319.2+/-60.1 microg/dL; P=0.004). Of nine patients with iron deficiency anemia (Hb <12 g/dL), eight patients had telogen hair loss (odds ratio: 10.5, 95%CI: 1.2-90.7; P=0.013). Odds ratio (95% confidence interval) for diffuse telogen hair loss was 21.0 (4.2-105.0) at serum ferritin levels < or =30 ng/mL. Women with iron deficiency status are at a risk of telogen hair loss. The important role of serum ferritin in hair loss is becoming more evident. In women without systemic inflammation or other underlying disorders, serum ferritin levels below or equal to 30 ng/mL are strongly associated with telogen hair loss. PMID- 20021983 TI - Genital contact allergy. AB - Irritant and allergic contact dermatitis is commonly seen in patients complaining of itching, burning and irritation in the genital area. The aim of this retrospective study was to establish the prevalence of allergic contact dermatitis patients with genital complaints. We followed 33 patients with persistent or recurrent genital redness, itching and burning sensation. Diagnosis was made by history, clinical examination and patch testing. Patch tests were carried out according to the International Contact Dermatitis Research Group with a standard series of allergens. We also tested topical pharmaceutical products that individual patients used for treating genital symptoms and patients self intimate hygiene products. There were 11 male and 22 female patients, mean age 38 years. Thirteen (39%) patients had one or more positive allergic reactions, mainly to nickel-sulfate, thimerosal, balsam of Peru, formaldehyde and neomycin sulfate. In seven of 13 patients with positive patch test results, these reactions were considered to be relevant to their clinical condition. Three patients had positive patch test reactions to their intimate hygiene products. One patient had positive patch test reaction to latex condom. Patients with genital symptoms are at a risk of developing contact sensitivity. Patch testing is useful in the management of these patients and many can be helped by allergen avoidance. PMID- 20021984 TI - Orofacial Crohn's disease: an oral enigma. AB - Crohn's disease is a chronic, relapsing, inflammatory disorder which may involve any segment of the bowel from mouth to anus. The mucocutaneous manifestations of Crohn's disease in the orofacial region are multiple, including oral Crohn's disease, metastatic Crohn's disease in sites noncontiguous with the bowel system, and reactive disorders such as pyoderma gangrenosum. Clinicians should be familiar with these extraintestinal manifestations and include this important and often serious disease in the evaluation of patients with selected orofacial disorders. The recognition of these manifestations may help prevent misdiagnosis and unnecessary treatment, and facilitates timely diagnosis, palliation and definitive therapy. PMID- 20021985 TI - Verrucous systemic lupus erythematosus. AB - Few patients with systemic lupus erythematosus have features of verrucous (hypertrophic) lupus erythematosus. A 29-year-old Caucasian woman with a 7-year history of systemic lupus erythematosus developed painful verrucous plaques on the nose. Erythematous, raised, indurated, hyperkeratotic plaques localized on the dorsa of the distal parts of the toes and over the interphalangeal joints of her fingers were also noted. A large, dull-red, indurated plaque with rolled borders on the bridge of the nose was most characteristic. Rapid therapeutic effect was obtained by systemic corticosteroid regimen. This verrucous variant of lupus erythematosus, sometimes clinically resembling actinic keratosis, keratoacanthoma and squamous cell carcinoma, is reviewed. PMID- 20021986 TI - Treatment of chronic urticaria. AB - Urticaria is a disorder characterized by rapid onset of localized swelling of the skin or mucosa, called wheals or urtica. According to frequency and duration, urticaria can be divided into acute and chronic type. Chronic urticaria is any type of urticaria occurring every day or twice per week, lasting longer than 6 weeks. Chronic urticaria is a common disorder and estimated prevalence is 1% of the population. Also, it is not rare in childhood. The pathogenesis of chronic urticaria has not yet been completely understood. Chronic urticaria is a heterogeneous group of disorders, and according to the etiology and cause, several groups of chronic urticaria are distinguished, i.e. autoimmune, pseudoallergic, infection-related, physical urticaria, vasculitis urticaria and idiopathic urticaria. Treatment and management of chronic urticaria can be non pharmacological and pharmacological, and sometimes it is not possible to control the disease with antihistamines only, which are considered to be the mainstay of treatment. In severe cases of chronic urticaria, especially if autoimmunity has been proven, several authors describe different modules of immunomodulation: cyclosporine, cyclophosphamide, mycophenolate-mofetil, omalizumab, plasmapheresis, systemic corticosteroids, and immunoglobulin therapy. This article primarily addresses the treatment of chronic idiopathic and autoimmune urticaria. PMID- 20021987 TI - Hypereosinophilic syndrome and proliferative diseases. AB - Therapy principles of the last decade and recent advances in the research of polynuclear eosinophil have led to a new approach in the hypereosinophilic syndrome (HES), with important consequences on the development of new and effective therapies. HES is defined by persistent and marked eosinophilia and eosinophil-related organ damage in the absence of any evident cause of hypereosinophilia. Two variants of HES have been characterized, with different prognosis and possible associations with malignant diseases such as myeloid leukemia or T-cell lymphomas. The lymphocytic variant of HES (L-HES) is characterized by the presence of T cell clones, IL-5 expression and possible progression to T-cell lymphoma. Besides steroid therapy, the anti-IL-5 monoclonal antibody mepolizumab is considered as a target therapy for L-HES. The myeloproliferative variant of HES (M-HES) is associated with an increased risk of myeloid leukemia and good response to anti-tyrosine-kinase therapy. The imatinib target is a kinase resulting from an 800-kb deletion on chromosome 4. The fusion gene Fip1-like 1-platelet-derived growth factor receptor alpha (FIP1L1-PDGFRA) has been validated as a marker of response to anti-tyrosine-kinase therapy. Early identification of HES variants is crucial for the rapid introduction of early and appropriately adjusted therapy. PMID- 20021988 TI - The preventive effect of calcitriol on skin squamous cell carcinoma may be due to its effect on prostaglandin E2 metabolism and biologic activity. PMID- 20021989 TI - First joint event between Europe and EADV: EADV-AAD Review Course State-of-the Art in Dermatology, Munich, 25-28 June 2009. PMID- 20021991 TI - Genetic association study of treatment response with olanzapine/fluoxetine combination or lamotrigine in bipolar I depression. AB - OBJECTIVE: To evaluate common genetic variations for association with symptomatic improvement in bipolar I depression following treatment with olanzapine/fluoxetine combination (OFC) or lamotrigine. METHOD: Symptom improvement was assessed in 88 OFC-treated and 85 lamotrigine-treated white patients with bipolar I depression in the 7-week acute period of a randomized, double-blind study comparing OFC (6/25, 6/50, 12/25, or 12/50 mg/d [olanzapine/fluoxetine]) with lamotrigine (titrated to 200 mg/d). The original study was conducted from November 2003 to August 2004. Single nucleotide polymorphisms (SNPs) were genotyped in a set of 19 candidate genes corresponding to known sites of activity for olanzapine and fluoxetine or previously associated with antidepressant or antipsychotic response. Primary outcome was the reduction in Montgomery-Asberg Depression Rating Scale (MADRS) total score as assessed by the difference by genotype from baseline to week 7 from a mixed-effects repeated measures analysis with terms for visit, genotype, genotype-by-visit interaction, and baseline MADRS score as a covariate. RESULTS: SNPs within the dopamine D(3) receptor and histamine H(1) receptor (HRH1) genes were significantly associated with response to OFC. SNPs within the dopamine D(2) receptor, HRH1, dopamine beta hydroxylase, glucocorticoid receptor, and melanocortin 2 receptor genes were significantly associated with response to lamotrigine. CONCLUSIONS: SNPs in specific candidate genes were associated with symptomatic improvement in a treatment-specific fashion. These results suggest the importance of dopaminergic effects in the treatment of patients with bipolar I depression and the potential utility of genotyping in selection of pharmacologic treatments for bipolar depression. PMID- 20021992 TI - Obesity and onset of significant depressive symptoms: results from a prospective community-based cohort study of older men and women. AB - OBJECTIVE: Although several cross-sectional studies have linked obesity and depression, less is known about their longitudinal association and about the relative influence of obesity subtypes. We prospectively examined whether obesity (specifically, abdominal) increased the risk of onset of depression in a population-based sample of older persons. METHOD: Participants were 2,547 nondepressed, well-functioning white and black persons, aged 70-79 years, enrolled in the Health, Aging, and Body Composition Study, an ongoing prospective community-based cohort study. Baseline measurements were conducted between April 1997 and June 1998. Overall obesity was assessed by body mass index (BMI) and percent body fat (measured by dual energy x-ray absorptiometry), whereas abdominal obesity measures included waist circumference, sagittal diameter, and visceral fat (measured by computer tomography). Onset of significant depressive symptoms was defined as a Center for Epidemiologic Studies Depression 10-item score > or = 10 at any annual follow-up over 5 years and/or new antidepressant medication use. Persistent depression was defined as depression at 2 consecutive follow-up visits. RESULTS: Over 5 years, significant depressive symptoms emerged in 23.7% of initially nondepressed persons. In men, both overall (BMI: hazard ratio [HR] per SD increase = 1.20; 95% CI, 1.03-1.40) and abdominal obesity (visceral fat: HR per SD increase = 1.19; 95% CI, 1.07-1.33) predicted onset of depressive symptoms after adjustment for sociodemographics. When BMI and visceral fat were adjusted for each other, only visceral fat was significantly associated with depression onset (HR = 1.18; 95% CI, 1.04-1.34). Stronger associations were found for persistent depressive symptoms. No associations were found in women. CONCLUSION: This study shows that obesity, in particular visceral fat, increases the risk of onset of significant depressive symptoms in men. These results suggest that specific mechanisms might relate visceral fat to the onset of depression. PMID- 20021993 TI - Does comorbid posttraumatic stress disorder affect the severity and course of psychotic major depressive disorder? AB - BACKGROUND: Major depressive disorder (MDD) and posttraumatic stress disorder (PTSD) are commonly comorbid conditions that result in greater severity, chronicity, and impairment compared with either disorder alone. However, previous research has not systematically explored the potential effects of the psychotic subtyping of MDD and comorbid PTSD. METHOD: The sample in this retrospective case control study conducted from December 1995 to August 2006 consisted of psychiatric outpatients with DSM-IV-diagnosed psychotic MDD with PTSD, psychotic MDD without PTSD, or nonpsychotic MDD with PTSD presenting for clinic intake. Clinical indices of severity, impairment, and history of illness were assessed by trained diagnosticians using the Structured Clinical Interview for DSM-IV Axis I Disorders supplemented by items from the Schedule for Affective Disorders and Schizophrenia. RESULTS: In terms of current severity and impairment, the psychotic MDD with PTSD (n = 34) and psychotic MDD only (n = 26) groups were similar to each other, and both tended to be more severe than the nonpsychotic MDD with PTSD group (n = 263). In terms of history of illness, the psychotic MDD with PTSD group tended to show greater severity and impairment relative to either the psychotic MDD only or nonpsychotic MDD with PTSD groups. Furthermore, the psychotic MDD with PTSD patients had an earlier time to depression onset than patients with either psychotic MDD alone or nonpsychotic MDD with PTSD, which appeared to contribute to the poorer history of illness demonstrated in the former group. CONCLUSIONS: Future research should explore the possibility of a subtype of psychotic depression that is associated with PTSD, resulting in a poorer course of illness. The current findings highlight the need for pharmacologic and psychotherapeutic approaches that can be better tailored to psychotic MDD patients with PTSD comorbidity. PMID- 20021994 TI - Family outcomes from a randomized control trial of relapse prevention therapy in first-episode psychosis. AB - OBJECTIVE: We have previously reported that our combined individual and family cognitive-behavioral therapy (CBT) relapse prevention therapy (RPT) was effective in reducing relapse rates compared to treatment as usual (TAU) within a specialist program for young, first-episode psychosis patients who had reached remission on positive symptoms. Here, we report the outcomes for family participants of DSM-IV-diagnosed first-episode psychosis patients recruited between November 2003 and May 2005 over a 2.5-year follow-up period. The primary hypothesis was that, compared to family members receiving TAU, family participants who received RPT would have significantly improved appraisals of stressors related to caregiving. Secondary hypotheses were that RPT would be associated with reduced expressed emotion and improved psychological distress. METHOD: Family members were assessed at baseline and at 7-month, 12-month, 18 month, 24-month, and 30-month follow-up on appraisal of caregiving, expressed emotion, and psychological distress using the Experience of Caregiving Inventory, The Family Questionnaire, and the General Health Questionnaire of 28 Items, respectively. The family component of RPT was based on family behavioral therapy for schizophrenia with a specific focus on psychoeducation and CBT for relapse prevention. RESULTS: Thirty-two families received RPT, and 31 families received TAU. There were significant group effects for aspects of the appraisal of caregiving, including negative symptoms, positive personal experiences, and total positive score on the Experience of Caregiving Inventory. Time effects were evident for emotional overinvolvement and for aspects of the appraisal of caregiving. There were no significant effects for psychological distress. CONCLUSIONS: The relatives of patients who received RPT perceived less stress related to their relative's negative symptoms and an increase in perceived opportunities to make a positive contribution to the care of their relative compared to carers in the TAU condition. Cognitive-behavioral therapy for relapse prevention showed promise in improving the experience of caregiving for family members of first-episode psychosis patients over a 2.5-year follow-up period. TRIAL REGISTRATION: anzctr.org.au Identifier: ACTRN12605000514606. PMID- 20021995 TI - Electroconvulsive therapy decreases dopamine D2receptor binding in the anterior cingulate in patients with depression: a controlled study using positron emission tomography with radioligand [11C]FLB 457. AB - OBJECTIVE: Electroconvulsive therapy (ECT) has been confirmed as one of the most effective treatments in drug-resistant major depression. However, the mechanism of ECT is still poorly understood. Although several lines of studies have focused on its effect on dopamine neurotransmission, the effects of ECT on dopamine D(2) receptors in a living human brain have not been investigated. Using positron emission tomography (PET) scans with the radioligand [(11)C]FLB 457, we aimed to evaluate the effect of ECT on extrastriatal D(2) receptor binding in medicated patients with major depressive disorder (MDD). METHOD: Seven patients with a DSM IV diagnosis of MDD underwent PET scans before and after a series of 6-7 treatments with bilateral ECT. Eleven healthy controls were scanned for comparison. All participants were scanned at the National Institute of Radiological Sciences, Chiba, Japan, between November 2000 and September 2005. The parametric images of [(11)C]FLB 457 binding were generated on the basis of a simplified reference tissue model. Voxel-based methods were used to assess the effect of ECT on D(2) receptor binding. RESULTS: There were no significant differences in D(2) receptor binding between patients with MDD and controls. All 7 patients showed clinical improvements in response to ECT treatment (P < .001). Significant changes in D(2) receptor binding, a mean of 25.2% reduction, were found in the right rostral anterior cingulate (AC) following ECT (P < .001). CONCLUSIONS: Electroconvulsive therapy decreased D(2) receptor binding in the rostral AC in MDD patients responding to ECT. Our finding suggests that one of the biologic mechanisms of ECT could be related to dopaminergic alteration in the rostral AC. PMID- 20021996 TI - Associations between serum lipids and major depressive disorder: results from the Netherlands Study of Depression and Anxiety (NESDA). AB - BACKGROUND: Several studies have suggested an association between lipids or lipoproteins and depression, but findings are contradictory. However, previous studies did not always take into consideration potentially mediating factors or heterogeneity of symptoms, which may clarify contradicting findings. METHOD: We compared levels of serum total, low-density lipoprotein (LDL), and high-density lipoprotein (HDL) cholesterol and triglyceride between 761 subjects with current major depressive disorder (MDD) (Composite International Diagnostic Interview, based on the DSM-IV), 1,071 subjects with remitted MDD, and 629 controls, aged 18 to 65 years. Subjects participated in the baseline assessment of the Netherlands Study of Depression and Anxiety, which lasted from September 2004 to February 2007. We studied the impact of adjustment for sociodemographics, lifestyle related covariates, and antidepressant use and examined the association between specific psychopathological characteristics and lipid/lipoprotein levels. RESULTS: HDL cholesterol level was lower (P = .007) and triglyceride level was higher (P = .001) in current MDD versus remitted MDD and controls. After adjustment for level of education, body mass index (BMI), smoking status, and alcohol use, dissimilarities lost statistical significance. Depression severity, comorbid dysthymia, and melancholic and atypical features were all associated with lipids/lipoproteins, but most associations attenuated after adjustment for covariates, especially BMI. The association between melancholic features and lower HDL cholesterol (P = .038) and between atypical depression and higher total and LDL cholesterol (P = .004 and P = .002, respectively) persisted after full adjustment. CONCLUSIONS: Adverse lipoprotein patterns were found in patients with MDD. The fact that these associations diminished after adjustment for lifestyle related factors, especially BMI, suggests that the unfavorable lipid/lipoprotein pattern among depressed subjects is mainly secondary to lifestyle-related factors. However, melancholic features were independently associated with lower HDL cholesterol, and atypical depression was independently associated with higher total and LDL cholesterol. PMID- 20021997 TI - Levetiracetam in generalized social anxiety disorder: a double-blind, randomized controlled trial. AB - OBJECTIVE: This multicenter, double-blind, placebo-controlled, 2-arm, parallel group study was carried out to determine the effectiveness and safety of the novel anticonvulsant levetiracetam for the treatment of generalized social anxiety disorder (GSAD). METHOD: After a 1-week, single-blind, placebo run-in period, 217 adult outpatients meeting DSM-IV criteria for social anxiety disorder, generalized type, were randomly assigned (1:1) to 12 weeks of double blind treatment with either levetiracetam (n = 111) or placebo (n = 106). Participants were required to have scores of >or= 60 on the Liebowitz Social Anxiety Scale (LSAS) and a total score of or= 30% reduction in LSAS score) were similar with 41.3% (levetiracetam) and 46.6% (placebo). No significant between group differences were found on secondary outcome measures, which included changes in Sheehan Disability Scale, Clinical Global Impression of Change, and HDRS scores. CONCLUSIONS: Although well-tolerated, levetiracetam failed to separate from placebo in this trial for the treatment of moderate to severe GSAD. PMID- 20021998 TI - Effect of viewing angle on arm reaching while standing in a virtual environment: potential for virtual rehabilitation. AB - Functional arm movements, such as reaching while standing, are planned and executed according to our perception of body position in space and are relative to environmental objects. The angle under which the environment is observed is one component used in creating this perception. This suggests that manipulation of viewing angle may modulate whole body movement to affect performance. We tested this by comparing its effect on reaching in a virtually generated environment. Eleven young healthy individuals performed forward and lateral reaches in the virtual environment, presented on a flat screen in third-person perspective. Participants saw a computer-generated model (avatar) of themselves standing in a courtyard facing a semi-circular hedge with flowers. The image was presented in five different viewing angles ranging from seeing the avatar from behind (0 degrees), to viewing from overhead (90 degrees). Participants attempted to touch the furthest flower possible without losing balance or stepping. Kinematic data were collected to analyze endpoint displacement, arm-postural coordination and center of mass (COM) displacement. Results showed that reach distance was greatest with angular perspectives of approximately 45-77.5 degrees , which are larger than those used in analogous real world situations. Larger reaches were characterized by increased involvement of leg and trunk body segments, altered inter-segmental coordination, and decreased inter-segmental movement time lag. Thus a viewing angle can be a critical visuomotor variable modulating motor coordination of the whole body and related functional performance. These results can be used in designing virtual reality games, in ergonomic design, teleoperation training, and in designing virtual rehabilitation programs that re-train functional movement in vulnerable individuals. PMID- 20022000 TI - Understanding emotional expression using prosodic analysis of natural speech: refining the methodology. AB - Emotional expression is an essential function for daily life that can be severely affected in some psychological disorders. Laboratory-based procedures designed to measure prosodic expression from natural speech have shown early promise for measuring individual differences in emotional expression but have yet to produce robust within-group prosodic changes across various evocative conditions. This report presents data from three separate studies (total N = 464) that digitally recorded subjects as they verbalized their reactions to various stimuli. Format and stimuli were modified to maximize prosodic expression. Our results suggest that use of evocative slides organized according to either a dimensional (e.g., high and low arousal - pleasant, unpleasant and neutral valence) or categorical (e.g., fear, surprise, happiness) models produced robust changes in subjective state but only negligible change in prosodic expression. Alternatively, speech from the recall of autobiographical memories resulted in meaningful changes in both subjective state and prosodic expression. Implications for the study of psychological disorders are discussed. PMID- 20021999 TI - Loss-of-function mutations in the PRPS1 gene cause a type of nonsyndromic X linked sensorineural deafness, DFN2. AB - We report a large Chinese family with X-linked postlingual nonsyndromic hearing impairment in which the critical linkage interval spans a genetic distance of 5.41 cM and a physical distance of 15.1 Mb that overlaps the DFN2 locus. Mutation screening of the PRPS1 gene in this family and in the three previously reported DFN2 families identified four different missense mutations in PRPS1. These mutations result in a loss of phosphoribosyl pyrophosphate (PRPP) synthetase 1 activity, as was shown in silico by structural analysis and was shown in vitro by enzymatic activity assays in erythrocytes and fibroblasts from patients. By in situ hybridization, we demonstrate expression of Prps1 in murine vestibular and cochlea hair cells, with continuous expression in hair cells and postnatal expression in the spiral ganglion. Being the second identified gene associated with X-linked nonsyndromic deafness, PRPS1 will be a good candidate gene for genetic testing for X-linked nonsyndromic hearing loss. PMID- 20022001 TI - A 3D-structural model of unsulfated chondroitin from high-field NMR: 4-sulfation has little effect on backbone conformation. AB - The glycosaminoglycan chondroitin sulfate is essential in human health and disease but exactly how sulfation dictates its 3D-structure at the atomic level is unclear. To address this, we have purified homogenous oligosaccharides of unsulfated chondroitin (with and without (15)N-enrichment) and analysed them by high-field NMR to make a comparison published chondroitin sulfate and hyaluronan 3D-structures. The result is the first full assignment of the tetrasaccharide and an experimental 3D-model of the hexasaccharide (PDB code 2KQO). In common with hyaluronan, we confirm that the amide proton is not involved in strong, persistent inter-residue hydrogen bonds. However, in contrast to hyaluronan, a hydrogen bond is not inferred between the hexosamine OH-4 and the glucuronic acid O5 atoms across the beta(1-->3) glycosidic linkage. The unsulfated chondroitin bond geometry differs slightly from hyaluronan by rotation about the beta(1-->3) psi dihedral (as previously predicted by simulation), while the beta(1-->4) linkage is unaffected. Furthermore, comparison shows that this glycosidic linkage geometry is similar in chondroitin-4-sulfate. We therefore hypothesise that both hexosamine OH-4 and OH-6 atoms are solvent exposed in chondroitin, explaining why it is amenable to sulfation and hyaluronan is not, and also that 4-sulfation has little effect on backbone conformation. Our conclusions exemplify the value of the 3D-model presented here and progress our understanding of glycosaminoglycan molecular properties. PMID- 20022002 TI - Efficient prediction methods for selecting effective siRNA sequences. AB - Although short interfering RNA (siRNA) has been widely used for studying gene functions in mammalian cells, its gene silencing efficacy varies markedly and there are only a few consistencies among the recently reported design rules/guidelines for selecting siRNA sequences effective for mammalian genes. Another shortcoming of the previously reported methods is that they cannot estimate the probability that a candidate sequence will silence the target gene. This paper first reviewed the recently reported siRNA design guidelines and clarified the problems concerning the guidelines. It then proposed two prediction methods-Radial Basis Function (RBF) network and decision tree learning-and their combined method for selecting effective siRNA target sequences from many possible candidate sequences. They are quite different from the previous score-based siRNA design techniques and can predict the probability that a candidate siRNA sequence will be effective. The methods imply high estimation accuracy for selecting candidate siRNA sequences. PMID- 20022003 TI - Skill sets and competencies for the modern military surgeon: lessons from UK military operations in Southern Afghanistan. AB - INTRODUCTION: British military forces remain heavily committed on combat operations overseas. UK military operations in Afghanistan (Operation HERRICK) are currently supported by a surgical facility at Camp Bastion, in Helmand Province, in the south of the country. There have been no large published series of surgical workload on Operation HERRICK. The aim of this study is to evaluate this information in order to determine the appropriate skill set for the modern military surgical team. METHOD: A retrospective analysis of operating theatre records between 1st May 2006 and 1st May 2008 was performed. Data was collated on a monthly basis and included patient demographics, operation type and time of operation. RESULTS: During the study period 1668 cases required 2210 procedures. Thirty-two per cent were coalition forces (ISAF), 27% were Afghan security forces (ANSF) and 39% were civilians. Paediatric casualties accounted for 14.7% of all cases. Ninety-three per cent of cases were secondary to battle injury and of these 51.3% were emergencies. The breakdown of procedures, by specialty, was 66% (1463) orthopaedic, 21% (465) general surgery, 6% (139) head and neck, 5% (104) burns surgery and a further 4% (50) non-battle, non-emergency procedures. There was an almost twofold increase in surgical workload in the second year (1103 cases) compared to the first year of the deployment (565 cases, p<0.05). DISCUSSION: Surgical workload over the study period has clearly increased markedly since the initial deployment of ISAF forces to Helmand Province. A 6 week deployment to Helmand Province currently provides an equivalent exposure to penetrating trauma as 3 years trauma experience in the UK NHS. The spectrum of injuries seen and the requisite skill set that the military surgeon must possess is outside that usually employed within the NHS. A number of different strategies; including the deployment of trainee specialist registrars to combat hospitals, more focused pre-deployment military surgery training courses, and wet laboratory work are proposed to prepare for future generations of surgeons operating in conflict environments. PMID- 20022004 TI - Perioperative and long-term complications among obese women undergoing vaginal surgery. AB - OBJECTIVE: To determine whether obesity influenced the risk of perioperative and long-term complications in patients undergoing vaginal surgery. METHODS: Women diagnosed with pelvic organ prolapse who underwent vaginal surgery between March 1999 and May 2007 were classified into 3 groups: normal weight (body mass index [BMI, calculated as weight in kilograms divided by the square of height in meters] 18.5-23.0); overweight (BMI 23.0-27.5); and obese (BMI >or=27.5). Complications were subdivided into perioperative (<1 month) and long-term (>or=1 month). Statistical analysis was performed using analysis of variance, chi(2) test, and logistic regression. RESULTS: Demographic information showed no differences among the groups, except for the presence of comorbidity (P=0.03). Blood loss and transfusion as a perioperative complication and urinary retention as a long-term complication were significantly different among the groups: odds ratio (OR) 2.46 (95% confidence interval [CI], 1.38-4.39; P<0.01); and OR 2.20 (95% CI, 1.21-4.03; P=0.03), respectively. CONCLUSION: Major complications were rare, and most were not significantly different among the groups. However, obesity was a protective factor against blood loss and transfusion, and long-term urinary retention. PMID- 20022005 TI - Effectiveness of an instructional DVD on third- and fourth-degree laceration repair for obstetrics and gynecology postgraduate trainees. AB - OBJECTIVE: To assess the effectiveness of an instructional DVD on the anatomy and repair of anal sphincter lacerations to improve postgraduate trainees' understanding. METHODS: A total of 71 obstetrics and gynecology trainees completed a pretest of third- and fourth-degree lacerations to assess baseline knowledge and perceptions. Question categories included anatomy, antibiotics, anesthesia, repair methods, complications, postoperative care, and risk factors. After 1 year of clinical experience, 67 trainees (94%) were randomly assigned into DVD (intervention) and non-DVD (control) groups. A post-test was administered 4 weeks later. RESULTS: In the DVD group (n=34), mean scores on the pretest versus the post-test were 65% vs 74% for postgraduate year (PGY)-1 (P=0.09); 72% vs 83% for PGY-2 (P=0.06); 67% vs 83% for PGY-3 (P=0.01); and 75% vs 87% for PGY-4 (P<0.001). In the non-DVD group (n=33), mean scores did not change significantly for any year level. The increase in score from pretest to post-test was significantly different between the 2 groups, independent of year (P<0.001). DVD group scores improved significantly over non-DVD group scores in anatomy (P=0.005) and repair methods (P=0.042) subscales. CONCLUSION: An educational video is an effective tool for improving understanding of third- and fourth-degree lacerations for physicians-in-training. PMID- 20022006 TI - Situated clinical encounters in the negotiation of religious and spiritual plurality: a critical ethnography. AB - BACKGROUND: Despite increasingly diverse, globalized societies, little attention has been paid to the influence of religious and spiritual diversity on clinical encounters within healthcare. OBJECTIVES: The purpose of the study was to analyze the negotiation of religious and spiritual plurality in clinical encounters, and the social, gendered, cultural, historical, economic and political contexts that shape that negotiation. DESIGN: Qualitative: critical ethnography. SETTINGS: The study was conducted in Western Canada between 2006 and 2009. Data collection occurred on palliative, hospice, medical and renal in-patient units at two tertiary level hospitals and seven community hospitals. PARTICIPANTS: Participants were recruited through purposive sampling and snowball technique. Twenty healthcare professionals, seventeen spiritual care providers, sixteen patients and families and twelve administrators, representing diverse ethnicities and religious affiliations, took part in the study. METHODS: Data collection included 65 in-depth interviews and over 150h of participant observation. RESULTS: Clinical encounters between care providers and recipients were shaped by how individual identities in relation to religion and spirituality were constructed. Importantly, these identities did not occur in isolation from other lines of social classification such as gender, race, and class. Negotiating difference was a process of seeing spirituality as a point of connection, eliciting the meaning systems of patients and creating safe spaces for the expression of that meaning. CONCLUSIONS: The complexity of religious and spiritual identity construction and negotiation raises important questions about language and about professional competence and boundaries in clinical encounters where religion and spirituality are relevant concerns. PMID- 20022007 TI - Attitudes of nursing staff towards electronic patient records: a questionnaire survey. AB - BACKGROUND: A growing number of health care organizations are implementing a system of electronic patient records (EPR). This implies a change in work routines for nursing staff, but it could also be regarded as an opportunity to improve the quality of care. OBJECTIVE: The objective of this paper is to obtain more insight into the usefulness of EPR as perceived by nursing staff and to clarify the determinants of nursing staff's acceptance of EPR. Determinants were tested using an extended version of the Technology Acceptance Model. DESIGN: Nursing staff members (NAs and RNs) completed a survey questionnaire about the use of EPR in health care, and their experiences, perceptions and attitudes regarding EPR. SETTINGS: All nursing staff members were working in Dutch hospitals, psychiatric organizations, care organizations for mentally retarded people, home care organizations, nursing homes or homes for the elderly. PARTICIPANTS: The study population is a nationally representative Dutch research sample, further referred to as the Nursing Staff Panel. The Panel consists of a permanent group of Nursing Assistants (NAs) and Registered Nurses (RNs), who are prepared to fill in a postal questionnaire twice a year on average. In January 2009, 685 participants completed the questionnaire. RESULTS: Nursing staff members associate EPR with improved care, especially qualitatively better and safer care. They also expect an increase in costs of care, while anticipating only a relatively small rise in the number of patients that can be cared for. In general, the effects of EPR on the work circumstances of nursing staff are expected to be negative. Job-related characteristics were found to be determinants of attitudes towards using EPR. A relatively positive attitude towards EPR was found in three categories of nursing staff in particular, i.e. staff working at least 30h per week, staff already using EPR and staff working in hospitals. Nursing staff in management positions also tend to have a more positive attitude. When the Technology Acceptance Model was tested, attitudes towards EPR were primarily associated with job-related characteristics and perceived usefulness with respect to quality of care. CONCLUSIONS: The implementation strategies for EPR need to take account of the job characteristics of the intended future users. If implementation is to be successful, it is important that the users understand the beneficial effects of EPR on the quality of care. PMID- 20022008 TI - A report on the Genetics of Complex Diseases meeting of the British Atherosclerosis Society, Cambridge, UK, 17-18 September 2009. PMID- 20022009 TI - Commentary to factors predicting cardiovascular events in statin-treated diabetic and non-diabetic patients with coronary atherosclerosis. PMID- 20022010 TI - The mechanical effects of different levels of cement penetration at the cement bone interface. AB - The mechanical effects of varying the depth of cement penetration in the cement bone interface were investigated using finite element analysis (FEA) and validated using companion experimental data. Two FEA models of the cement-bone interface were created from micro-computed tomography data and the penetration of cement into the bone was varied over six levels each. The FEA models, consisting of the interdigitated cement-bone constructs with friction between cement and bone, were loaded to failure in tension and in shear. The cement and bone elements had provision for crack formation due to excessive stress. The interfacial strength showed a strong relationship with the average interdigitation (r(2)=0.97 and r(2)=0.93 in tension and shear, respectively). Also, the interface strength was strongly related with the contact area (r(2)=0.98 and r(2)=0.95 in tension and shear, respectively). The FEA results compared favorably to the stiffness-strength relationships determined experimentally. Overall, the cement-bone interface was 2.5 times stronger in shear than in tension and 1.15 times stiffer in tension than in shear, independent of the average interdigitation. More cracks occurred in the cement than in the bone, independent of the average interdigitation, consistent with the experimental results. In addition, more cracks were generated in shear than in tension. In conclusion, achieving and maintaining maximal infiltration of cement into the bone to obtain large interdigitation and contact area is key to optimizing the interfacial strength. PMID- 20022011 TI - Two methods for the separation of monounsaturated octadecenoic acid isomers. AB - The identification and quantification of complex mixtures of cis and trans octadecenoic (18:1) fatty acid isomers presents a major challenge for conventional one-dimensional GC/FID analysis of their methyl esters. We have compared the use of two methods to achieve optimized separations of positional and geometrical octadecenoic fatty acid isomers-comprehensive two-dimensional gas chromatography (GCxGC), and silver ion high performance liquid chromatography interfaced to atmospheric pressure photoionization (APPI) mass spectrometry. Nine isomers of octadecenoic acid methyl ester were well separated on a single silver ion column with a mobile phase of 0.018% acetonitrile and 0.18% isopropanol in hexane. Reproducible retention times were obtained with relative standard deviations of around 1% over 5 injections. The extra selectivity and reproducibility afforded by APPI-MS, together with the wide separation of cis and trans isomers by silver ion chromatography, resulted in a promising method for measurement of octadecenoic acid FAME. The GCxGC separation was performed using various column combinations, and optimal separation was obtained by coupling an ionic liquid column (Supelco SLB-IL100 [1,9-di(3-vinyl-imidazolium) nonane bis(trifluoromethyl) sulfonyl imidate]) in the first dimension with a SGE BPX50 (50% phenyl polysilphenylene-siloxane) in the second dimension. These methods have been applied to the analysis of octadecenoic acid in milk and beef fat. PMID- 20022012 TI - Factors affecting atrazine concentration and quantitative determination in chlorinated water. AB - Although the herbicide atrazine has been reported to not react measurably with free chlorine during drinking water treatment, this work demonstrates that at contact times consistent with drinking water distribution system residence times, a transformation of atrazine can be observed. Some transformation products detected through the use of high performance liquid chromatography-electrospray mass spectrometry are consistent with the formation of N-chloro atrazine. The effects of applied chlorine, pH, and reaction time on the transformation reaction were studied to help understand the practical implications of the transformation on the accurate determination of atrazine in drinking waters. The errors in the determination of atrazine are a function of the type of dechlorinating agent applied during sample preparation and the analytical instrumentation utilized. When a reductive dechlorinating agent, such as sodium sulfite or ascorbic acid is used, the quantification of the atrazine can be inaccurate, ranging from 2-fold at pH 7.5 to 30-fold at pH 6.0. The results suggest HPLC/UV and ammonium chloride quenching may be best for accurate quantification. Hence, the results also appear to have implications for both compliance monitoring and health effects studies that utilize gas chromatography analysis with sodium sulfite or ascorbic acid as the quenching agent. PMID- 20022013 TI - Design and application of Hadamard-injectors coupled with gas and supercritical fluid sample collection systems in Hadamard transform-gas chromatography/mass spectrometry. AB - A novel Hadamard transform-gas chromatography/mass spectrometry (HT-GC/MS) system equipped with on-line sample collection systems is described. A Hadamard-injector was successfully designed and then coupled with an on-line adsorption/desorption system for detecting volatile organic compounds (VOCs) and a supercritical fluid extraction (SFE) system, respectively, by HT-GC/MS. Six VOCs and three pesticides were used as model compounds. In the former case, an activated-charcoal trap was used to trap VOCs from the indoor air. After 10L of indoor air had passed through the trap, the condensed components were heated and simultaneously injected into the GC column through the Hadamard-injector, based on Hadamard codes. In a second experiment, a sample of rice was spiked with three types of pesticides and the sample then extracted using a commercially available supercritical fluid extractor. After extraction, the extracted components were transferred to a holding tank and simultaneously injected into the GC column also using the Hadamard-injector. The findings show that, in both cases, the combination of on line sample collection methods and the use of the Hadamard transform resulted in improved sensitivity and detection. Compared to the single injection used in most GC/MS systems, the signal-to-noise (S/N) ratios were substantially improved after inverse Hadamard transformation of the encoded chromatogram. PMID- 20022014 TI - Identification/quantification of multiple pesticide residues in food plants by ultra-high-performance liquid chromatography-time-of-flight mass spectrometry. AB - In this study, the potential of ultra-high-performance liquid chromatography coupled with the time-of-flight mass spectrometry (UHPLC-TOF MS) to enable rapid and comprehensive analysis of 212 pesticide residues in QuEChERS extracts obtained from four plant matrices has been investigated. Method optimization is discussed in detail. In addition to molecular adducts, also fragment ions were provided for all target pesticides, thus obtaining at least three identification points required by European Decision 2002/657/EC was achieved. To get maximum information on analytes present in the extracts, each sample was examined within two injections, the first in a positive and the next one in a negative ionization mode. Under UHPLC conditions, both analyses were completed within 24min. For more than 96% of pesticides involved in this study, the limit of quantification was < or =10micro/kg. As a part of the work, strategy enabling screening of non-target pesticides and their metabolites is demonstrated on analysis of real-life samples. PMID- 20022015 TI - Understanding and mitigating conductivity transitions in weak cation exchange chromatography. AB - Large conductivity fluctuations were observed during a high pH wash step in a weak cation exchange chromatography process. These conductivity transitions resulted in a conductivity drop during pH increase and a conductivity rise during pH decrease. In some cases, the absolute conductivity change was greater than 6mS/cm which was sufficient to affect target protein retention on the column. Further investigation revealed that wash buffer concentration, resin ligand density, and resin ligand pK have a profound effect on the magnitude of the conductivity transitions and the shape of corresponding pH traces. A potentiometric electrode selective for sodium ions was used to measure effluent counterion concentrations from two preparative resins during high pH washes, and the number of exchangeable counterions was compared to predictions made using ion exchange equilibrium theory. Results from this analysis show that conductivity transitions can be effectively mitigated without compromising process performance by optimizing the trade-off between wash buffer concentration and wash phase duration. PMID- 20022016 TI - Supercritical fluid extraction: Recent advances and applications. AB - Among the different extraction techniques used at analytical and preparative scale, supercritical fluid extraction (SFE) is one of the most used. This review covers the most recent developments of SFE in different fields, such as food science, natural products, by-product recovery, pharmaceutical and environmental sciences, during the period 2007-2009. The revision is focused on the most recent advances and applications in the different areas; among them, it is remarkable the strong impact of SFE to extract high value compounds from food and natural products but also its increasing importance in areas such as heavy metals recovery, enantiomeric resolution or drug delivery systems. PMID- 20022017 TI - Selection of suitable operating conditions to minimize the gradient equilibration time in the separation of drugs by Ultra-High-Pressure Liquid Chromatography with volatile (mass spectrometry-compatible) buffers. AB - Reversed phase gradient elution is the method of choice for pharmaceuticals analysis since it allows reducing the analysis time while improving both the quality of the separation and the detection limits. The current trends are towards faster separations which can be achieved thanks to equipments withstanding ultra-high pressures and/or high temperatures. Under such conditions, gradient separations can be carried out within a few minutes or even a few tens of seconds. A long equilibration time in addition to the gradient time can be therefore very detrimental. In this work, we investigated the extent to which the gradient equilibration time can be reduced and which parameters mainly affect the retention variability of ionizable compounds when using volatile buffers. We first found out an excellent repeatability between run-to-run experiments whatever the equilibration time and the operating conditions. We then pointed out the key operating parameters which allow achieving reproducible runs when varying the equilibration time between runs. With a view of reducing the equilibration time, the effects of various conditions were examined. The latter include the type of additive for mobile phase pH adjustment, the initial eluent composition, the type of stationary phase, the temperature and the flow-rate. Although much remains to be understood about the equilibration process, our study allows making progress in the knowledge of this phenomenon. Based on the present results, a beneficial effect of both temperature and flow-rate was highlighted and operating conditions leading to faster column equilibration are suggested. PMID- 20022018 TI - Simultaneous determination of four 5-hydroxy polymethoxyflavones by reversed phase high performance liquid chromatography with electrochemical detection. AB - Accumulating evidence has suggested the potential health-promoting effects of 5 hydroxy polymethoxyflavones (5-OH-PMFs) naturally existing in citrus genus. However, research efforts are hampered by the lack of reliable and sensitive methods for their determination in plant materials and biological samples. Using reversed-phase high performance liquid chromatography (HPLC) equipped with electrochemical (EC) detection, we have developed a fast and highly sensitive method for quantification of four 5-OH-PMFs, namely 5-hydroxy-6,7,8,3',4' pentamethoxyflavone, 5-hydroxy-3,6,7,8,3',4'-hexamethoxyflavone, 5-hydroxy-6,7,4' trimethoxyflavone, and 5-hydroxy-6,7,8,4'-tetramethoxyflavone. The method was fully validated in terms of linearity, accuracy and precision. The limit of detection (LOD) was determined as being between 0.65 and 1.8ng/mL (ppb), demonstrating an over 160 times higher sensitivity in comparison with the previously reported method using UV detection. The recovery rate of the method was between 96.17% and 110.82%, and the precision for the retention times and peak areas was all below 13%. The method was successfully used to quantify 5-OH PMFs with a wide range of abundance in the citrus products and preparations, such as orange juice, citrus peel, and dried tangerine peel. The quantification method for 5-OH-PMFs developed herein could be useful for the nutritional and pharmacological studies of these compounds in future. PMID- 20022019 TI - Microwave-assisted phase-transfer catalysis for the rapid one-pot methylation and gas chromatographic determination of phenolics. AB - Microwave-assisted phase-transfer catalysis (PTC) is reported for the first time, for the one-step extraction-derivatization-preconcentration and gas chromatographic determination of twenty phenols and ten phenolic acids. The well established phase-transfer catalytic methylation is largely accelerated when heating is replaced with the "greener" microwave irradiation. The overall procedure was thoroughly optimized and the analytes were determined by GC/MS. The method presented adequate analytical characteristics being more sensitive in analyzing phenols than phenolic acids. The limits of detection without any additional preconcentration steps (e.g. solvent evaporation) were adequate and ranged from 0.4 to 15.8ng/mL while limits of quantitation were between 1.2 and 33.3ng/mL. The method was applied to the determination of phenols, in spiked environmental samples and phenolic acids in aqueous infusions of commercially available pharmaceutical dry plants. The recoveries of fortified composite lake water samples and Mentha spicata aqueous infusions ranged from 89.3% to 117.3% for phenols and 93.3% to 115.2% for phenolic acids. PMID- 20022020 TI - Constructions of silver nanowires and copper oxide microrings by a surface formation technique. AB - We demonstrate a simple method to synthesize silver wires by thermal reduction of aqueous AgNO(3) droplet with catalytic anatase TiO(2) nanoparticles which were spin-coated on Si wafer. Structural characterization of the silver wires shows that the nanowires grow primarily along the [011] direction. SEM image of the silver wires clearly shows the catalytic TiO(2) nano-cluster attached to the end of the each silver wire. Since the process was surfactant-free, the silver nanowires prepared by our method retain the excellent electrical conductivity. The intrinsic resistivity calculated from the current-voltage curve for a wire with 12880.41 nm(2) cross-section area was 18.72 microohm cm, which is about 11.6 times higher than that of bulk silver (1.61 microohm cm). Our simple method can be also applied to generate CuO with ring-shaped microstructure by using ITO conducting glass as matrix. We have found that the size and reproducibility are well-controllable. A single phase of CuO ring-shaped microstructure with outer diameters ranging from approximately 13 to 17 microm and inner diameters ranging from approximately 1.4 to 3.3 microm was obtained. The composition of CuO microring was confirmed by thin-film XRD and XPS analyses. PMID- 20022021 TI - Improved enzymatic activity of Thermomyces lanuginosus lipase immobilized in a hydrophobic particulate mesoporous carrier. AB - Lipase from Thermomyces lanuginosus has been immobilized within particulate mesoporous silica carriers, with either hydrophilic or hydrophobic supporting surfaces, produced by the newly developed emulsion and solvent evaporation (ESE) method. The Michaelis-Menten model was used to calculate the parameters related to the enzymatic activity of lipase i.e. the turnover number, k(cat), and the specific activity. The specific activity was improved by immobilization of lipase onto the hydrophobic support, compared to lipase immobilized onto the hydrophilic support and lipase free in solution. The enhanced enzymatic activity of lipase onto a hydrophobic support was attributed to interfacial activation of the Thermomyces lanuginosus lipase when it is attached to a hydrophobic surface and a reduced denaturation. Confocal scanning laser microscopy (CLSM) studies, of fluorescently tagged lipase, showed that leakage of the lipase from the mesoporous particles was limited to an initial period of only a few hours. Both the rate and the amount of lipase leached were reduced when the lipase was immobilized onto the hydrophobic support. PMID- 20022022 TI - In situ precipitation of magnetic fluid encapsulated in giant liposomes. AB - It is well known that adding salt to a colloidal solution stabilized by electrostatic repulsions can induce its destabilization. When this colloidal solution is encapsulated inside liposomes an in situ precipitation can be induced by slightly modification of the environment: application of a magnetic field in the case of magnetic particles or increase of the temperature. It has been shown in a previous study that magnetic liposomes exhibit strong deformation under magnetic field. In this case the precipitation can be induced after their elongation under magnetic field and liposomes keep their shape even if the magnetic field is cut off. The following study is motivated by the desire to use the salt effect to precipitate magnetic nanoparticles inside giant liposomes. PMID- 20022023 TI - Extrapontine myelinolysis as presenting manifestation of adrenal failure: a case report. AB - BACKGROUND: Hyponatremia is a fairly common metabolic disorder. Hyponatremic myelinolysis is a relatively rare, life threatening complication with poorly understood pathophysiology, varied clinical manifestations and uncertain treatment. This case report highlights the range of clinical and imaging phenomena associated with hyponatremic myelinolysis. METHODS: Case report. RESULT: A middle aged lady presented with an acute delirious state, hypotension and severe hyponatremia on a background of skin hyper-pigmentation and weight loss. Her clinical course evolved to an akinetic-rigid state and later to parkinsonism. Extensive investigations for recognizing a primary neurologic disorder, including brain MRI and CSF analysis were normal, though she had disseminated miliary tuberculosis involving multiple organs. Brain MRI changes characteristic of extrapontine myelinolysis appeared two weeks after the onset of symptoms. The patient recovered completely over several weeks. CONCLUSION: This case of hyponatremic extrapontine myelinolysis occurred as the presenting manifestation of adrenal failure secondary to disseminated tuberculosis. Extraponine myelinolysis is difficult to diagnose in the context of delayed brain MRI changes and can have a favorable outcome with modern management. PMID- 20022024 TI - Neuronal and glial tau pathology in early frontotemporal lobar degeneration-tau, Pick's disease subtype. AB - Frontotemporal lobar degeneration-tau, Pick's disease subtype (PiD) is one of the major types of frontotemporal dementia, but its pathogenesis and disease mechanisms remain unclear. Here, we report a case of very early PiD. The patient was a 63-year-old healthy woman without dementia or any apparent psychosis. She was admitted to the hospital with multiple organ failure, and died three days later. The brain weighed 1050g and showed focal atrophy of the parahippocampal gyrus and right medial temporal lobe. Microscopically, neuronal loss and gliosis were limited to the atrophic areas. Surprisingly, Pick bodies (PiBs) and ballooned neurons were abundant throughout the bilateral temporal cortices, including the dentate gyrus. Cortical lamination of PiBs was predominant in the upper layer (layer II>VI), and the size of early PiBs tended to be smaller than that in severely affected areas. Numerous glial tau-positive inclusions (astrocytic inclusions, oligodendroglial coiled bodies, and threads) were found not only in the cerebral cortex but also in the temporal white matter. The neuropathological findings in this case suggest that PiB formation started long before the appearance of clinical symptoms and that PiB formation originating from small neurons may differ from other tau aggregations such as neurofibrillary tangles. PMID- 20022025 TI - Diagnostic evaluation of children with daytime incontinence. AB - PURPOSE: This article is one of the standardization documents of the International Children's Continence Society, and discusses how anatomical/iatrogenic and functional/urodynamic causes of daytime incontinence in children of all ages are to be diagnosed, how neurogenic bladder dysfunction or urinary tract infection is excluded as a cause of the wetting, and how further diagnostic evaluation of children with disturbances such as overactive bladder, voiding postponement and dysfunctional voiding is performed. The roles of history taking (including prenatal and perinatal issues and family history), physical examination, diagnostic bladder diaries, noninvasive urodynamic investigations and radiological imaging are delineated but therapy is not within the scope of this document. MATERIALS AND METHODS: This document was designed and written by an international panel of authors with a large experience in assessment of children with incontinence. RESULTS: The best evidence was retrieved from the literature and assembled in a standardization document. CONCLUSIONS: Assessment of children with daytime symptoms is discussed. A noninvasive approach in these children allows us to select patients who will need a more invasive assessment. PMID- 20022026 TI - Improvement in vesicoureteral reflux grade on serial imaging predicts resolution. AB - PURPOSE: When children are initially diagnosed with vesicoureteral reflux most undergo a period of antibiotic prophylaxis followed by serial imaging. Although improvement in reflux grade through time presumably predicts eventual resolution, the significance of changing grade through time is unknown. We examined whether improvement in reflux on serial imaging predicts resolution. MATERIALS AND METHODS: We retrospectively reviewed 1,761 children diagnosed with vesicoureteral reflux, of whom 965 had a minimum of 2 years of followup. We examined initial reflux grade and grade on serial imaging up to 5 years after the original diagnosis. For each child it was determined whether reflux was resolved, eventually resolved or never resolved. Groups were further stratified by clinical characteristics. RESULTS: Multivariate analysis revealed that male gender (HR 1.33, p = 0.05), age younger than 1 year at diagnosis (HR 1.35, p = 0.004), lower grade at presentation (grade I HR 2.2, grade II HR 1.96, grade III HR 1.33; p <0.001) and unilateral reflux (HR 1.39, p = 0.001) were all independent predictors of reflux resolution. Multivariate analysis also showed that reflux improvement on imaging 1 year after diagnosis (HR 3.14, p <0.0001) and improvement from the previous year at any point during followup (HR 1.8, p = 0.009) were independent predictors of reflux resolution. CONCLUSIONS: Consistent with previous findings, male gender, lower reflux grade at presentation, age less than 1 year at presentation and unilateral reflux were all predictive of reflux resolution. Our analysis also demonstrated that improvement in reflux grade on imaging study 1 year after diagnosis was predictive of resolution, and that reflux improvement from the previous year at any point during followup was an independent predictor of resolution. This information will prove valuable in clinical counseling and therapeutic decision making. PMID- 20022027 TI - Editorial comment. PMID- 20022028 TI - Next generation biodegradable ureteral stent in a yucatan pig model. AB - PURPOSE: Ureteral stents are commonly used to facilitate kidney drainage but they may produce significant stent symptoms and morbidity, and require a secondary procedure for removal. Previous biodegradable stents showed bio-incompatibility or inconsistent degradation, requiring extra procedures to remove undegraded stent fragments. We previously reported a first generation biodegradable stent composed of suture-like material that required placement through the lumen of a sheath and degraded by 10 weeks. We now report second and third generation biodegradable stents that degrade more rapidly and can be placed directly over a polytetrafluoroethylene guidewire. MATERIALS AND METHODS: Two groups of 16 Yucatan pigs each were unilaterally stented endoscopically with a control nondegradable (biostable) stent or a second generation degradable Uriprene stent. Blood studies, renal ultrasound and excretory urography were done throughout the study to determine renal function, hydronephrosis and stent degradation. Genitourinary organs were harvested at necropsy for pathological analysis. A third generation stent designed to improve degradation time was bilaterally implanted endoscopically into 4 Yorkshire Farm pigs (total of 8 stents), followed by excretory urography weekly to assess degradation and kidney function. Biomaterial parameters were tested. RESULTS: Second generation stents began degrading at 2 weeks and were completely degraded by 10 weeks. All third generation stents were degraded by 4 weeks. Hydronephrosis was considerably less in the Uriprene group than in control biostable stented kidneys. Biostable stented ureters showed an average higher degree of inflammation, uropathy and nephropathy. Physical characteristics indicate that Uriprene stents are significantly more resistant to stent compression and have markedly higher tensile strength and coil strength comparable to that of other commercially available plastic stents. CONCLUSIONS: Our study confirms that Uriprene stents are biocompatible and provide good renal drainage. They hold promise for decreasing the need for a secondary procedure and stent related morbidity, such as infection and irritative symptoms. PMID- 20022029 TI - Mucinous tubular and spindle cell carcinoma of the kidney. PMID- 20022031 TI - Computerized tomography of Fournier's Gangrene. PMID- 20022032 TI - Adaptive spaced education improves learning efficiency: a randomized controlled trial. AB - PURPOSE: Spaced education is a novel form of online education that harnesses the 2 psychology research findings of spacing and testing effects. Spaced education is delivered by daily emails containing clinically relevant multiple choice questions. To take advantage of the spacing effect the questions are repeated at fixed intervals for a fixed number of repetitions. An adaptive spaced education system was developed to customize spacing intervals and the number of repetitions based on learner knowledge level. To determine whether this system improves learning efficiency I performed a randomized trial to compare the learning efficiency of adaptive vs nonadaptive spaced education systems among surgery students at 2 medical schools. MATERIALS AND METHODS: A total of 62 year 3 students were randomized to identical course content in adaptive or nonadaptive spaced education formats. The course consisted of 40 validated, spaced education items on the 4 urology topics benign prostatic hyperplasia, erectile dysfunction, prostate cancer and prostate specific antigen screening. The nonadaptive cohort received daily emails containing 2 questions with a linear review of the material 20 days after initial presentation. The adaptive cohort received daily emails via an adaptive algorithm that limited the repetition of mastered content. Each cohort completed a validated end of course test. RESULTS: The adaptive cohort answered significantly fewer spaced education items than the nonadaptive cohort (p = 0.001) but achieved comparable end of course test scores (p = 0.37). The adaptive algorithm increased learning efficiency by 38%. CONCLUSIONS: Adaptive spaced education boosts learning efficiency. PMID- 20022033 TI - Improvement in neurogenic detrusor overactivity by peripheral C fiber's suppression with cyclooxygenase inhibitors. AB - PURPOSE: Cyclooxygenase inhibitors decrease micturition frequency in animals with bladder inflammation but to our knowledge the influence of cyclooxygenase inhibitors on detrusor overactivity has not been investigated. We evaluated the effects, and the site and mechanism of action of cyclooxygenase inhibitors on detrusor overactivity induced by cerebral infarction. MATERIALS AND METHODS: Cerebral infarcted rats underwent cumulative intravenous administration of the selective cyclooxygenase-1 inhibitor SC-560 (Sigma), the selective cyclooxygenase 2 inhibitor rofecoxib (Kemprotec, Middlesbrough, United Kingdom) or the nonselective cyclooxygenase inhibitor FYO-750 hourly plus a single intravenous administration of SC-560, rofecoxib or SC-560 plus rofecoxib. To evaluate the site of action cerebral infarcted rats underwent single intracerebroventricular or intrathecal administration of FYO-750. To evaluate the mechanism of action FYO 750 was intravenously administered in diuretic rats or cerebral infarcted rats pretreated with resiniferatoxin. RESULTS: For cumulative administration SC-560 (0.3 mg/kg), rofecoxib (0.3 mg/kg) and FYO-750 (0.1 to 1 mg/kg) significantly increased bladder capacity. For single administration neither SC-560 (0.03 mg/kg) nor rofecoxib (0.03 mg/kg) affected bladder capacity but SC-560 plus rofecoxib significantly increased bladder capacity vs vehicle. Intracerebroventricular and intrathecal administration of FYO-750 did not affect bladder capacity. FYO-750 did not affect urinary production in diuretic rats and the effects of FYO-750 were blocked by resiniferatoxin except at the highest drug dose. CONCLUSIONS: Results indicate that cyclooxygenase inhibitors improve detrusor overactivity caused by cerebral infarction by suppressing peripheral C fiber's without affecting urinary production. The nonselective cyclooxygenase inhibitor showed more potent efficiency than the selective cyclooxygenase-1 or the cyclooxygenase 2 inhibitor alone. PMID- 20022036 TI - Editorial comment. PMID- 20022034 TI - Bladder dysfunction and altered somatic sensitivity in PACAP-/- mice. AB - PURPOSE: PACAP and receptors are expressed in micturition pathways. Studies show that PACAP has a role in detrusor smooth muscle contraction to facilitate adenosine triphosphate release from urothelium and PACAP antagonism decreases cyclophosphamide induced bladder hyperreflexia. MATERIALS AND METHODS: PACAP contributions to micturition and somatic sensation were studied in PACAP knockout (PACAP(-/-)), litter mate heterozygote (PACAP(+/-)) and WT mice by conscious cystometry with continuous intravesical saline or acetic acid (0.5%) instillation, urination patterns, somatic sensitivity testing of hind paw and pelvic regions with calibrated von Frey filaments, and morphological bladder assessments. RESULTS: PACAP(-/-) mice had an increased bladder mass with fewer but larger urine spots. In PACAP(-/-) mice the lamina propria and detrusor smooth muscle were significantly thicker but the urothelium was unchanged. PACAP(-/-) mice had increased bladder capacity, voided volume and intercontraction interval with significantly increased detrusor contraction duration and large residual volume. WT mice responded to acetic acid (0.5%) with a decrease in voided volume and intercontraction interval but PACAP(+/-) and PACAP(-/-) mice did not respond. PACAP(-/-) mice were less responsive to somatic stimulation. PACAP(+/-) mice also had bladder dysfunction, and somatic and visceral sensory abnormalities but to a lesser degree. CONCLUSIONS: PACAP gene disruption contributes to changes in bladder morphology and function, and somatic and visceral hypoalgesia. PMID- 20022037 TI - New tissue bulking agent (polyacrylate polyalcohol) for treating vesicoureteral reflux: preliminary results in children. AB - PURPOSE: We report the preliminary results of endoscopic treatment of vesicoureteral reflux in children using polyacrylate polyalcohol copolymer. MATERIAL AND METHODS: We performed a prospective multicenter review of pediatric patients treated with subureteral injection of a new nonabsorbable substance. Only patients with 1 year of followup were included. RESULTS: A total of 83 patients underwent injection of polyacrylate polyalcohol copolymer at our institutions between 2005 and 2006. Among this group 18 males and 43 females with a median age of 58 months (range 9 months to 18 years) completed 1 year of followup. Reflux was bilateral in 27 patients (44.3%) and unilateral in 34 (55.7%). Number of injected ureters was 88. Reflux grade was V in 3 ureters (3.4%), IV in 12 (13.6%), III in 41 (46.6%) and II in 32 (36.4%). Mean +/- SD injected volume per unit was 0.76 +/- 0.43 ml. Median followup was 20 months (range 16 to 24). Complications after injection included dysuria in 6 patients (9.8%), fever in 3 (4.9%) and lumbar pain in 4 (6.6%). Reflux was eliminated in 78 renal units (88.6%), decreased to grade I in 6 (6.8%) and persisted in 4 (4.5%). Ureteral obstruction developed in 1 patient and was treated operatively. Overall success rate was 83.6%. CONCLUSIONS: Polyacrylate polyalcohol copolymer can be used to treat vesicoureteral reflux with comparable efficacy to other substances currently used, with a low rate of complications. PMID- 20022038 TI - Editorial comment. PMID- 20022039 TI - Febrile urinary tract infection after ureteroneocystostomy: a contemporary assessment at a single institution. AB - PURPOSE: Febrile urinary tract infection represents significant morbidity in patients with vesicoureteral reflux, especially following open surgical or endoscopic treatment. The reported incidence of febrile urinary tract infection after ureteroneocystostomy varies from 10% to 24%. We investigated the incidence of febrile urinary tract infection following ureteroneocystostomy in a contemporary, single institution series. MATERIALS AND METHODS: We retrospectively reviewed medical records of 395 consecutive patients undergoing ureteroneocystostomy for primary vesicoureteral reflux at our institution between 2002 and 2007. We examined demographic, diagnostic and operative data, including presence of postoperative febrile urinary tract infection. A Cox proportional hazards model was performed to assess predictors of febrile urinary tract infection following ureteroneocystostomy. RESULTS: Ureteroneocystostomy was performed in 395 patients (673 ureters) at a mean age of 58 months. The most common reflux grade was III (41%). The incidence of postoperative febrile urinary tract infection was 4.6% at a mean followup of 15 months. Postoperative dysfunctional elimination syndrome was a significant predictor of febrile urinary tract infection (HR 3.8, 95% CI 1.2-12, p = 0.02), and was identified in 58 of 340 toilet trained children (15% overall). Age at diagnosis, initial presentation, age at surgery, indication for surgery, reflux grade, laterality, surgical technique and preoperative dysfunctional elimination syndrome were not predictive of postoperative febrile urinary tract infection. CONCLUSIONS: The incidence of febrile urinary tract infection following ureteroneocystostomy may be lower than previously reported. The presence of postoperative dysfunctional elimination syndrome is a significant predictor of postoperative febrile urinary tract infection. PMID- 20022040 TI - Editorial comment. PMID- 20022041 TI - Primary bladder neck dysfunction in children and adolescents III: results of long term alpha-blocker therapy. AB - PURPOSE: Primary bladder neck dysfunction is a nonneurogenic voiding disorder frequently overlooked in pediatrics. The diagnosis classically is made by videourodynamics but can also be made with noninvasive uroflow studies with pelvic floor electromyography. We report our long-term results using alpha blocker therapy in patients with primary bladder neck dysfunction. MATERIALS AND METHODS: We reviewed 51 neurologically normal children (mean age 11.6 years, range 3.5 to 17.8) meeting criteria for primary bladder neck dysfunction who underwent alpha-blocker therapy for at least 1 year. All patients were symptomatic with abnormal flow parameters and an electromyogram lag time of 6 seconds or more on initial uroflow/electromyography. Pretreatment and on treatment uroflow/electromyogram studies were performed in all patients. Average and maximum uroflow rates, electromyogram lag times and post-void residual volumes were compared. RESULTS: After a mean followup of 46.2 months (range 12 to 124) mean average and maximum uroflow rates improved from 7.0 to 12.4 cc per second and from 12.4 to 20.3 cc per second, respectively, while mean electromyogram lag time decreased from 30.8 to 5.8 seconds (all p <0.01). Of the patients 85% reported subjective symptomatic relief. A total of 15 patients (29%) stopped alpha-blocker therapy for various reasons, none related to side effects. Repeat off-treatment uroflow/electromyogram studies showed that measured parameters reverted to pretreatment values (all p <0.05). Eight of these 15 patients eventually resumed alpha-blocker therapy, while only 3 remained asymptomatic off of the alpha-blocker. CONCLUSIONS: alpha-Blocker therapy continues to benefit children with primary bladder neck dysfunction even after 3 years of treatment. Few patients can come off of alpha-blocker therapy without returning to their pretreatment state, suggesting the condition is likely chronic in most patients. PMID- 20022044 TI - Role of TREK-1 potassium channel in bladder overactivity after partial bladder outlet obstruction in mouse. AB - PURPOSE: Mouse models of partial bladder outlet obstruction cause bladder hypertrophy. Expression of a number of ion channels is altered in hypertrophic detrusor muscle, resulting in bladder dysfunction. We determined whether mechanosensitive TREK-1 channels are present in the murine bladder and whether their expression is altered in partial bladder outlet obstruction, resulting in abnormal filling responses. MATERIALS AND METHODS: Partial bladder outlet obstruction was surgically induced in CD-1 mice and the mice recovered for 14 days. Cystometry was done to evaluate bladder pressure responses during filling at 25 microl per minute in partial bladder outlet obstruction mice and sham operated controls. TREK-1 channel expression was determined at the mRNA and protein levels by quantitative reverse transcriptase-polymerase chain reaction and Western blotting, respectively, and localized in the bladder wall using immunohistochemistry. RESULTS: Obstructed bladders showed about a 2-fold increase in weight vs sham operated bladders. TREK-1 channel protein expression on Western blots from bladder smooth muscle strip homogenates was significantly decreased in obstructed mice. Immunohistochemistry revealed a significant decrease in TREK-1 channel immunoreactivity in detrusor smooth muscle in obstructed mice. On cystometry the TREK-1 channel blocker L-methioninol induced a significant increase in premature contractions during filling in sham operated mice. L methioninol had no significant effect in obstructed mice, which showed an overactive detrusor phenotype. CONCLUSIONS: TREK-1 channel down-regulation in detrusor myocytes is associated with bladder overactivity in a murine model of partial bladder outlet obstruction. PMID- 20022045 TI - Editorial comment. PMID- 20022046 TI - Parental satisfaction after open versus robot assisted laparoscopic pyeloplasty: results from modified Glasgow Children's Benefit Inventory Survey. AB - PURPOSE: Since its inception, robot assisted laparoscopic pyeloplasty has rapidly become the minimally invasive surgical intervention of choice for treating ureteropelvic junction obstruction at our institution. The large initial investment in robot assisted surgery is frequently justified by its association with improved optics and instrument articulation, decreased postoperative pain, shorter length of hospitalization and improved cosmesis. However, there are no data specifically showing patient satisfaction with robot assisted laparoscopic pyeloplasty compared to traditional open surgery. MATERIALS AND METHODS: A previously validated satisfaction survey (Glasgow Children's Benefit Inventory) with 14 additional questions specifically addressing postoperative satisfaction was mailed to all parents (as patient proxy) of children who had undergone open or robot assisted laparoscopic pyeloplasty between January 2006 and December 2008. RESULTS: A total of 78 parents responded (response rate 70%). All responses achieving statistical significance favored robot assisted laparoscopic pyeloplasty. Parents of children who underwent robot assisted laparoscopic pyeloplasty reported significantly higher satisfaction with "overall life," confidence, self-esteem, burden of postoperative followup and size of incision scar. CONCLUSIONS: Parent satisfaction was greater with robot assisted laparoscopic pyeloplasty than with open surgery regarding amount of cosmesis and recovery. Interestingly the differences in satisfaction were not as large as anticipated, suggesting the impact of confounding factors such as age and preoperative parental expectations. Future large-scale prospective studies using validated surveys specific to pediatric surgery are needed to elucidate further the true benefits of minimally invasive surgical technology such as robot assistance. PMID- 20022047 TI - Validation of a high fidelity adult ureteroscopy and renoscopy simulator. AB - PURPOSE: Surgical simulation technology may efficiently train and accurately assess the acquisition of many skills. Surgical simulators often lack realism and can be expensive at $3,000 to more than $60,000. We assessed the face, content and construct validity of a newly developed, anatomically accurate, reasonably priced high fidelity ureteroscopy and renoscopy trainer. MATERIALS AND METHODS: A total of 46 participants, including attending urologists, urology residents, medical students and industry representatives, assessed the face and content validity of the simulator using a standard questionnaire. Ten experienced ureteroscopists with greater than 30 procedures per year and 10 novice ureteroscopists with 0 were assessed on the ability to perform flexible ureteroscopy, renoscopy and intrarenal basket extraction of a lower pole calculus using the adult ureteroscopy trainer (Ideal Anatomic Modeling, Holt, Michigan). Subject performance was assessed by an experienced ureteroscopist using a checklist, global rating scale and time to task completion. RESULTS: Of participants 100% rated the trainer as realistic and easy to use, and thought it was a good training tool, 98% thought that it would serve as a good training format and 96% would recommend it to urology trainees. All participants recommended it for use in residency programs and 96% would or would have used it during residency. Only 37.5% vs 100% of experienced vs novice ureteroscopists would use it to practice. Of participants 9% foresaw a problem with the trainer. On the trainer experienced ureteroscopists scored significantly higher on the global rating scale (mean +/- SD 33.1 +/- 1.3 vs 15.0 +/- 2.7, p <0.0001) and checklist (4.1 +/- 1.0 vs 2.4 +/- 1.1, p = 0.004), and required less time to complete the task (141.2 +/- 40.1 vs 447.2 +/- 301.7 seconds, p = 0.01). CONCLUSIONS: Our preliminary study suggests the face, content and construct validity of the adult ureteroscopy trainer as a high fidelity ureteroscopy and renoscopy trainer. PMID- 20022048 TI - Testicular asymmetry and adolescent varicoceles managed expectantly. AB - PURPOSE: Adolescent varicocele is often associated with testicular asymmetry. Depending on the degree of asymmetry, some physicians will recommend surgery. However, given the possibility that asynchronous growth may be transient, others advocate for a period of observation. We reviewed our outcomes in such patients who were initially treated expectantly. MATERIALS AND METHODS: We retrospectively reviewed our pediatric varicocele database. We analyzed the outcomes of patients presenting for evaluation of varicocele who were followed with serial testicular volume measurements using scrotal ultrasound or ring orchidometry and who had at least a 6-month interval between measurements. Fisher's exact test was used to compare groups based on initial and final testicular asymmetry. RESULTS: We identified 181 patients (median age 13.8 years) who were followed expectantly. Serial volume measurements had been obtained at a median interval of 12 months (interquartile range 8 to 21) between first and most recent visits. Mean percent asymmetry for the group did not change with time. Among patients who initially had less than 20% asymmetry 35% had 20% or greater asymmetry on followup, and among those with 20% or greater asymmetry initially 53% remained in that range (p = 0.007). CONCLUSIONS: Asymmetry can be a transient phenomenon. Patients with initial asymmetry can end up with significant asymmetry, and many with significant asymmetry can have catch-up growth. However, when patients have a peak retrograde flow of 38 cm per second or greater on duplex Doppler ultrasound in association with 20% or greater asymmetry spontaneous catch-up growth is unlikely to occur. PMID- 20022049 TI - Polymorphisms of the TNF-alpha and ACE genes, and renal scarring in infants with urinary tract infection. AB - PURPOSE: Susceptibility to renal scarring is increasingly investigated through polymorphisms of genes regulating inflammation and fibrosis. TNF-alpha and ACE gene polymorphisms have been studied in chronic renal conditions but their role in urinary tract infection and vesicoureteral reflux associated renal scarring is unclear. We investigated the relationship between TNF-alpha A/G and ACE I/D polymorphisms, and renal scarring after urinary tract infection in infants. MATERIALS AND METHODS: ACE I/D and TNF-alpha -308 A/G polymorphisms were investigated with restriction fragment length polymorphism analysis in 39 boys and 25 girls with a first urinary tract infection before age 2 years and in 77 controls. Genotype and allele frequencies were compared among children with urinary tract infection with and without renal scarring, and controls. RESULTS: ACE I/D genotype frequencies were similar among infants with urinary tract infection with and without renal scarring, and controls. However, all 6 children with severe renal scarring and impaired renal function bore a D allele, 5 of which were DD homozygotes. D allele was more common in these severely affected children than in their peers with urinary tract infection and mild or no renal scarring (OR 9.92, 95% CI 1.24-79, p = 0.012), and controls (OR 8.03, 95% CI 1.01 64, p = 0.029). No differences were observed in TNF-alpha A/G genotype frequencies among the 3 groups. Presence of vesicoureteral reflux was not related to phenotypes or allele frequencies. CONCLUSIONS: Our findings suggest that D allele polymorphism of the ACE gene is associated with urinary tract infection related severe renal scarring in young children. PMID- 20022050 TI - Editorial comment. PMID- 20022052 TI - Urinary flow patterns in first year of life. AB - PURPOSE: We studied the natural development of urinary flow and lower urinary tract function in healthy male infants. MATERIALS AND METHODS: Custom-made ultrasound flow probes connected to a flowmeter were mounted on the penis in 20 infants who previously had been assessed in the immediate postnatal period. Median subject age was 10.7 months (range 9.2 to 19.8). Flow data were sampled to a personal computer and flow curves were assessed regarding configuration, maximum flow rate and voided volume. Results were analyzed statistically and were compared to those obtained in the neonatal period. Comparison of uroflow parameters was assessed by using analysis of variance, while contingency coefficients and Wilcoxon test were used for comparisons involving nominal and paired data, respectively. A p value of less than 0.05 was considered statistically significant. RESULTS: A total of 19 infants had evaluable data, of whom 15 also had evaluable data from the original neonatal study. Flow curve pattern was bell shaped in 32% of patients, interrupted in 46%, staccato in 15%, tower in 3% and spike-dome in 3%. Dyscoordinated patterns accounted for 46% of all flows, a significant increase compared to the neonatal period, in which only 34% of flows were considered dyscoordinated (p <0.01). While voided volume increased significantly with age, maximum flow rate remained more or less stable. CONCLUSIONS: Contrary to conventional wisdom, infants continue to exhibit urinary flow dyscoordination to an even greater extent than in the neonatal period. Therefore, the anticipated normalization of urinary flow is most likely to occur after the first year of life. PMID- 20022053 TI - Bladder dysfunction in a new mutant mouse model with increased superoxide--lack of nitric oxide? AB - PURPOSE: Nitric oxide mediates urethral smooth muscle relaxation and may also be involved in detrusor activity control. Mice with mutation in the Immp2l gene have high superoxide ion levels and a consequent decrease in the bioavailable amount of nitric oxide. We studied bladder function in this mouse model. MATERIAL AND METHODS: Young male mutants at ages 4 to 6 months, old female mutants at age 18 months and healthy WT age matched controls were used. The detrusor contractile response to carbachol and electrical field stimulation was tested in isolated detrusor strips in organ baths. In vivo bladder function was evaluated by cystometry in conscious animals. RESULTS: Young male mutants had significantly lower micturition and higher post-void residual volume than WT controls. They had pronounced voiding difficulty and strained when initiating micturition. Detrusor contractile responses to carbachol and electrical field stimulation were similar in mutant and WT mice. Old female mutant mice had lower bladder capacity and micturition volume, and higher micturition frequency and bladder-to-body weight ratio than WT controls. In the in vitro study detrusor strips from mutants showed a lower maximum response to carbachol. CONCLUSIONS: Mice with mutation in the Immp2l gene have bladder dysfunction, mainly characterized by emptying abnormalities in young males and increased detrusor activity in old females. Detrusor function was preserved in young males and impaired in old females. These animals are a natural model of oxidative stress with low bioavailable nitric oxide. Thus, they are interesting tools in which to evaluate the role of these conditions on bladder dysfunction. PMID- 20022054 TI - The miR-17-92 cluster is over expressed in and has an oncogenic effect on renal cell carcinoma. AB - PURPOSE: miRNAs are small, nonprotein coding RNAs that are differentially expressed in many malignancies. We previously identified 80 miRNAs that are dysregulated in clear cell renal cell carcinoma. In this study we validated over expression of the miR-17-92 cluster in clear cell renal cell carcinoma and tested the effect of 2 members of this cluster (miR-17-5p and miR-20a) on tumor proliferation. We also elucidated the role of miRNA in clear cell renal cell carcinoma pathogenesis with bioinformatics. MATERIALS AND METHODS: miRNA expression was validated by quantitative reverse transcriptase-polymerase chain reaction. The cell proliferation effect of miR-17-5p and miR-20a was tested in a renal adenocarcinoma cell line model. Multiple in silico analyses were done of dysregulated miRNAs. RESULTS: We validated miR-71-92 cluster over expression in clear cell renal cell carcinoma by quantitative reverse transcriptase-polymerase chain reaction. Transfection of miR-20a inhibitor significantly decreased cell proliferation in a dose dependent manner. Transfection of miR-17-5p, which is not endogenously expressed in the ACHN cell line, led to increased cell proliferation compared to control values. This effect was suppressed by miR-17-5p inhibitor. Bioinformatics analysis identified 10 clusters of miRNAs dysregulated in clear cell renal cell carcinoma that followed the same expression patterns. We also identified matching patterns between reported chromosomal aberration in clear cell renal cell carcinoma and miRNA dysregulation for 37.5% of the miRNAs. Target prediction analysis was done using multiple algorithms. Many key molecules in clear cell renal cell carcinoma pathogenesis, including HIFs, mTOR, VEGF and VHL, were potential targets for dysregulated miRNAs. CONCLUSIONS: A significant number of dysregulated proteins in clear cell renal cell carcinoma are potential miRNA targets. Also, many clear cell renal cell carcinoma dysregulated miRNAs are phylogenetically conserved. PMID- 20022055 TI - Editorial comment. PMID- 20022056 TI - Long-term results of bulking agent injection for persistent incontinence in cases of neurogenic bladder dysfunction. AB - PURPOSE: We evaluated the effectiveness of bladder neck injection as a supplementary treatment for persistent low pressure incontinence after unsatisfactory fascial sling procedures in patients with neurogenic lower urinary tract dysfunction. MATERIALS AND METHODS: A total of 89 patients with neurogenic lower urinary tract dysfunction underwent fascial sling procedures between 1992 and 2005. Because of unsatisfactory results, 27 patients received endoscopic injection of a bulking agent. All patients included in the study underwent urodynamic examination after the sling procedure, which revealed persistent low pressure transurethral leakage of urine. We retrospectively analyzed the endoscopic approach used to administer the bladder neck injection, method of postoperative catheterization and number of injections given. Efficacy of bladder neck injection was graded by the patient and the urologist. RESULTS: After a median followup of 8 years (range 2.5 to 14) only 2 patients (7%) were continent after having received a single injection of bulking agent. A total of 12 patients (44%) were given a second injection and 8 (30%) were given a third injection but these subsequent injections did not result in continence. Of the patients 16 (59%) eventually underwent bladder neck surgery, 2 (7%) were dry and 8 (30%) accepted the inconvenience. Two patients underwent ileocystoplasty and 1 patient underwent botulinum A toxin (Botox(R)) injection due to decreased bladder capacity and poor bladder compliance. Neither the endoscopic approach nor the method of postoperative catheterization affected the success rate. CONCLUSIONS: Bladder neck injection after failure of primary sling procedures has limited value in patients with neurogenic lower urinary tract dysfunction. Repeat bladder neck injection yields no additional benefits. PMID- 20022057 TI - Re: Sequential sorafenib and sunitinib for renal cell carcinoma M. P. Sablin, S. Negrier, A. Ravaud, S. Oudard, C. Balleyguier, J. Gautier, C. Celier, J. Medioni and B. Escudier J Urol 2009; 182: 29-34. PMID- 20022058 TI - Ethylene glycol induced hyperoxaluria increases plasma and renal tissue asymmetrical dimethylarginine in rats: a new pathogenetic link in hyperoxaluria induced disorders. AB - PURPOSE: The pathogenesis of kidney stones remains elusive. There is some evidence that hyperoxaluria may effect vascular endothelium and many studies link renal stones to atherosclerosis. Also, renal vascular endothelial cells regulate proximal tubular epithelial cell function. We determined the effect of hyperoxaluria on plasma and tissue levels of asymmetrical dimethylarginine. The secondary aim was to determine the effect of verapamil on asymmetrical dimethylarginine. MATERIALS AND METHODS: A total of 42 Sprague-Dawley rats were included in the study. In groups 1A, 1B and 1C hyperoxaluria was induced with ethylene glycol for 2 weeks. Groups 2A, 2B and 2C received ethylene glycol for 14 days and verapamil for 28 days. Control group 3 received no specific medication but distilled water. Blood samples were obtained at 24 hours and at study end, and kidney samples were obtained at 24 hours, and 7 and 28 days for histopathological evaluation. RESULTS: Plasma asymmetrical dimethylarginine increased early in the hyperoxaluric group (p = 0.0002). The effect was retained at the end of the study period (p = 0.01). There was no increase in asymmetrical dimethylarginine in the verapamil group on short-term and long-term followup. Hyperoxaluria induced a significantly dense staining pattern in renal tissue asymmetrical dimethylarginine vs controls (p = 0.01). Asymmetrical dimethylarginine staining did not differ in the control and verapamil groups. CONCLUSIONS: Increased systemic and local tissue asymmetrical dimethylarginine may help explain the pathogenetic mechanisms of hyperoxaluria induced disorders such as nephrolithiasis and atherosclerosis. PMID- 20022059 TI - Novel excitatory effects of adenosine triphosphate on contractile and pacemaker activity in rabbit urethral smooth muscle. AB - PURPOSE: Adenosine triphosphate is thought to be an important neurotransmitter in urethral smooth muscle but its physiological role is still unclear. We characterized the effects of adenosine triphosphate on contractile and pacemaker activity in rabbit urethral smooth muscle. MATERIALS AND METHODS: Tension recordings were made from strips of rabbit proximal urethral smooth muscle. Membrane currents from freshly isolated smooth muscle cells and interstitial cells of Cajal were recorded using the patch clamp technique. Intracellular Ca(2+) was measured using confocal microscopy. RESULTS: Exogenous application of adenosine triphosphate (10 microM) evoked robust contractions that were inhibited by the type 2 purinergic receptor blocker suramin (100 microM) and the selective type 2 purinergic Y1 receptor antagonist MRS2500 (Tocris Bioscience, Ellisville, Missouri) (100 nM). Application of the type 2 purinergic Y receptor agonist 2 MeSADP (1 microM) mimicked the effects of adenosine triphosphate. When smooth muscle cells were studied under voltage clamp at -60 mV, adenosine triphosphate evoked a large single inward current (greater than 1.2 nA) but 2-MeSADP produced a small current (about 16 pA). In contrast, when interstitial cells of Cajal were held at -60 mV, they showed spontaneous transient inward currents that were increased in frequency by adenosine triphosphate and 2-MeSADP. These excitatory effects were inhibited by suramin and MRS2500. Interstitial cells of Cajal showed spontaneous Ca(2+) waves that were increased in frequency by adenosine triphosphate and 2-MeSADP. These effects were also inhibited by suramin and MRS2500. CONCLUSIONS: Contractile effects of adenosine triphosphate in urethral smooth muscle are mediated by the activation of type 2 purinergic Y receptors on interstitial cells of Cajal. PMID- 20022060 TI - In vitro, ex vivo and in vivo isotherms for renal cryotherapy. AB - PURPOSE: Preoperative planning for renal cryotherapy is based on isotherms established in gel. We replicated gel isotherms and correlated them with ex vivo and in vivo isotherms in a porcine model. MATERIALS AND METHODS: PERC-17 CryoProbes (1.7 mm) and IceRods (1.47 mm) underwent trials in gel, ex vivo and in vivo porcine kidneys. Temperatures were recorded at 13 predetermined locations with multipoint thermal sensors. RESULTS: At the cryoprobe temperatures were not significantly different along the probe in any medium for either system (p = 0.0947 to 0.9609). However, away from the probe ex vivo and in vivo trials showed warmer temperatures toward the cryoprobe tip for each system (p = 0.0003 to 0.2141). Mean +/- SE temperature 5 mm distal to the cryoprobe tip in vivo was 19.2C +/- 16.1C for CryoProbes and 27.3C +/- 11.2C for IceRods. Temperatures were consistently colder with CryoProbes than with IceRods in gel (p <0.00005), ex vivo (p <0.00005) and in vivo (p = 0.0014). At almost all sites temperatures were significantly colder in gel and in ex vivo kidney than in in vivo kidney for CryoProbes (p = 0.0107 and 0.0008, respectively) and for IceRods (each p <0.00005). CONCLUSIONS: Gel and ex vivo isotherms do not predict the in vivo pattern of freezing. Thus, they should not be used for preoperative planning. The cryoprobe should be passed 5 mm beyond the tumor border to achieve suitably cold temperatures. Multipoint thermal sensor probes are recommended to record actual temperature during renal cryotherapy. PMID- 20022061 TI - [Diffuse large B cell lymphoma associated with hemophagocytic syndrome]. PMID- 20022062 TI - [Typical and atypical pneumonia: Clinical usefulness?]. PMID- 20022063 TI - [Comparison of equations to estimate the glomerular filtration in people aged 75 or older without known renal disease]. AB - BACKGROUND AND OBJECTIVE: To compare the glomerular filtration rate (GFR) of patients aged 75 and older without known renal disease estimated by the MDRD-4 IDMS (MDRD) formula, urine 24h creatinine clearance (24h-CC) and Cockroft-Gault (CG) formula. PATIENTS AND METHODS: There were included 70 persons aged 75 and older from the geriatric external consultation. We compared the MDRD against the CG and the 24h-CC. RESULTS: Means of GFR were: 24h-CC: 56,60 + or - 22,79 ml/min/1.73 m(2) and CG: 54,27 + or - 15,25 ml/min/1.73 m(2), significantly lower than with MDRD: 69,78 + or - 18,53 ml/min/1.73 m(2) (p<0.001). Age was correlated with the difference between CG and MDRD (coefficient of correlation: 0,59; r(2):0,34; p<0,001). Moderate agreement was obtained between MDRD with CG (k coefficient = 0.37; CI95%=0.19-0.55) and MDRD with 24-CC (k coefficient = 0.39; CI95% = 0.22-0.57). CONCLUSIONS: The MDRD and CG formulas to estimate the FGR in people aged 75 and older are not interchangeable. There are needed specific studies in old people to establish the most precise formula. PMID- 20022064 TI - [Heart failure, anemia and angiodysplasia: what else can we do?]. PMID- 20022065 TI - Relationship between increased arterial stiffness and other markers of target organ damage. AB - BACKGROUND AND OBJECTIVES: The purpose of the present study was to assess the relationship of arterial stiffness with other markers of target organ damage, and the clinical factors related to it. PATIENTS AND METHODS: Cross-sectional study that included 208 (115 men) never treated hypertensive, non-diabetic patients (mean age, 49+/-12 years). In addition to a full clinical study, 24h ambulatory blood pressure (BP), and determination of left ventricular hypertrophy (LVH) and microalbuminuria were performed. Clinical arterial stiffness was assessed by carotid-femoral pulse wave velocity (PWV) obtained with applanation tonometry (SphygmoCor-System). RESULTS: PWV was 8.3 (7.3-9.9)m/s (median, interquartile range). Stepwise regression analysis revealed that age (beta=0.086, p<0.001), 24 h pulse pressure (beta=0.058, p<0.001), and low-density lipoprotein (LDL) cholesterol (beta=0.009, p<0.013) were independent determinants of PWV. PWV>12m/s (indicating target organ lesion) was present in only 16 (7.7%) patients, less frequent than LVH (28% of the patients) and microalbuminuria (16%). However, of the 16 patients with elevated PWV, 10 (62%) had neither LVH or microalbuminuria. In a logistic multivariate regression analysis the factors related to elevated PWV were age > or =45 in man and > or =55 in women (OR: 23.8, 95% CI: 2.7-195.5; p=0.004), LDL cholesterol > or =160mg/dl (OR: 10.6, 95% CI: 2.6-42.7; p=0.001) and increased 24-h pulse pressure > or =55mmHg (OR: 3.9, 95% CI: 1.2-12.9; p=0.03). CONCLUSIONS: In untreated middle age hypertensives arterial stiffness assessed by PWV is less frequent than LVH or microalbuminuria. PWV is mainly related to age, LDL cholesterol, and pulse pressure values. PMID- 20022066 TI - [On Raynaud's phenomenon]. PMID- 20022067 TI - [Pharmacologic measures for the prevention of mechanical ventilation-associated pneumonia]. AB - Ventilator associated pneumonia is the principal infectious complication in the Intensive Care Unit (ICU), and represents the main infectious cause of morbidity and mortality. Its diagnosis and management is complex. Consequently, its prevention becomes a cornerstone in daily clinical practice. Daily interruption of sedation, oral and digestive decontamination, prophylactic administration of systemic and/or inhaled antibiotics, glycemic control, stress ulcer prophylaxis, transfusion policy and timing and adequacy of antibiotic treatment are the main suggested pharmacologic interventions. The aim of this review is to critically describe the principal pharmacologic interventions for the prevention of ventilator associated pneumonia, focusing on the degree of the evidence and the appropriateness for daily clinical practice. PMID- 20022068 TI - [Philosophy, medicine and clinical reasoning]. PMID- 20022069 TI - [The role of bisphosphonates of adjuvant therapy in breast cancer]. AB - Breast cancer is the most common neoplasm in women. Bone is the most common site of metastatic spread from primary operable breast cancer, causing pain, fractures and hypercalcemia. This spread depends on the release of osteolytic substances by the cancer cells. The osteoclasts also release growth factors that can act back on the cancer cells to activate growth. Biphosphonates, antiosteolytic agents, have been shown to reduce the progression of established bone metastases. Emerging evidence suggests that biphosphonates also have antitumor and antimetastatic properties, including the inhibition of angiogenesis, tumor-cell invasion, and adhesion in bone; the induction of apoptosis; antitumor sinergy with cytotoxic chemotherapy; and immunomodulatory effects through induction of gamma/delta T cells. These findings were the background and rationale for ongoing clinical trials, in order to establish their role as a part of adjuvant treatment for early breast cancer. The oral biphosphonate clodronate (1600mg/d) is effective in patients with bone metastases. When used as adjuvant therapy, given to patients with operable breast cancer for two years, clodronate has been reported to reduce the risk of bone metastases during a 2-year study period with a significant reduction in mortality. This benefit supports its use as additional adjuvant therapy for patients with operable breast cancer. Zoledronic acid, a potent nitrogen-containing biphosphonate, has been shown to maintain or increase bone mineral density (BMD) in premenopausal women with early-stage breast cancer receiving adjuvant hormone therapies as well as healthy postmenopausal women with low BMD. Recent large clinical trials with biphosphonates in the (neo)adjuvant setting for early-breast cancer, demonstrate an improvement of disease-free and overall survival as well as increased pathologic rate responses. PMID- 20022070 TI - [Argon plasma coagulation therapy of upper esophageal web in a patient with Plummer-Vinson syndrome: a new therapeutical option]. PMID- 20022071 TI - Chemical and functional changes of human insulin by in vitro incubation with blood from diabetic patients in oxidative stress. AB - Oxidative stress damage to biomolecules has been implicated in several diseases including diabetes mellitus. In the present study, we investigated the effect of oxidative stress in whole blood (WB) from diabetic patients (n = 60) on recombinant human insulin. Insulin was incubated with WB obtained from diabetic patients (DP) who had hyperglycemia (>300 mg/dL) or from 41 healthy volunteers (HV). Whole blood of DP, unlike WB of HV, induced higher values of formazan (142%), dityrosines (279%), and carbonyls (58%) in the insulin residues. Interestingly, the insulin modified by WB of DP showed less hypoglycemic activity in rat (30%) in comparison with insulin incubated with WB of HV. The incubation of insulin in WB from DP induces chemical changes in insulin and a decrease in its biological activity, events that might be associated with the high levels of oxidative stress markers found in the plasma of these patients. PMID- 20022072 TI - High-saturated-fat diet induces gestational diabetes and placental vasculopathy in C57BL/6 mice. AB - Gestational diabetes mellitus (GDM) is a commonly encountered disorder of mid to late pregnancy that is characterized by hyperglycemia, hyperinsulinemia, and impaired glucose tolerance. Gestational diabetes mellitus is thought to be multifactorial in origin and derives from both genetic and environmental factors. However, the mechanisms underlying GDM are incompletely understood; and current GDM animal models do not appear to closely mimic the clinical situation in humans. The present study used environmental exposure to high-saturated-fat diet (HFD) in an effort to develop a GDM mouse model that closely simulates metabolic abnormalities seen in human GDM. This model was then used to determine the contributions of HFD-induced placental oxidative stress (OS) and vascular dysregulation, which are observed in GDM patients and are believed to contribute to the pathogenesis of the disease. Pathologic manifestations of the disease were quantified by evaluating the extent of placental lipid peroxidation and by determining protective effects of dietary antioxidant quercetin supplementation to reduce HFD-associated placental OS. In this study, female C57BL/6 mice were fed HFD for 1 month before conception and throughout gestation to mimic chronic maternal fast food consumption. Maternal body weight increased from gestation day (GD) 0 to GD19 by 41% with HFD, as compared with 23% in control dams; HFD dams also developed insulin resistance (66% increase in plasma insulin and 27% increase in plasma glucose levels by GD10) as compared with control dams. Placentas from HFD GD19 dams demonstrated loss of trophoblasts and OS-mediated labyrinthine endothelial cellular damage, the latter of which was prevented with quercetin supplementation. Our findings suggest that prenatal HFD alters glucose metabolism and elevates placental OS, which are believed to collectively relate to improper formation of the conceptus and impaired birth outcome. PMID- 20022074 TI - WITHDRAWN PMID- 20022073 TI - [Staging rectal cancer]. AB - Rectal cancer has a high incidence in our area. The different treatment options that can be used in each case depend fundamentally on the extension of the tumor at the time of diagnosis. Thus, radiologists have a central role in the management of this disease. We discuss the optimum imaging technique for staging rectal cancer, some fundamental histological aspects, and the accepted semiology in the interpretation of different imaging studies carried out in these patients, as well as the diagnostic performance of each technique. We provide a general overview of the prognosis and different treatment options. This knowledge is very useful for radiologists participating in multidisciplinary teams and for understanding the transcendence of the information provided by image interpretation in these patients. PMID- 20022075 TI - Emergence of a group 3 coronavirus through recombination. AB - Analyses of turkey coronavirus (TCoV), an enteric disease virus that is highly similar to infectious bronchitis virus (IBV) an upper-respiratory tract disease virus in chickens, were conducted to determine the adaptive potential, and genetic changes associated with emergence of this group 3 coronavirus. Strains of TCoV that were pathogenic in poults and nonpathogenic in chickens did not adapt to cause disease in chickens. Comparative genomics revealed two recombination sites that replaced the spike gene in IBV with an unidentified sequence likely from another coronavirus, resulting in cross-species transmission and a pathogenicity shift. Following emergence in turkeys, TCoV diverged to different serotypes through the accumulation of mutations within spike. This is the first evidence that recombination can directly lead to the emergence of new coronaviruses and new coronaviral diseases, emphasizing the importance of limiting exposure to reservoirs of coronaviruses that can serve as a source of genetic material for emerging viruses. PMID- 20022076 TI - Degradation mechanism of alachlor during direct ozonation and O(3)/H(2)O(2) advanced oxidation process. AB - The degradation of alachlor by direct ozonation and advanced oxidation process O(3)/H(2)O(2) was investigated in this study with focus on identification of degradation byproducts. The second-order reaction rate constant between ozone and alachlor was determined to be 2.5+/-0.1M(-1)s(-1) at pH 7.0 and 20 degrees C. Twelve and eight high-molecular-weight byproducts (with the benzene ring intact) from alachlor degradation were identified during direct ozonation and O(3)/H(2)O(2), respectively. The common degradation byproducts included N-(2,6 diethylphenyl)-methyleneamine, 8-ethyl-3,4-dihydro-quinoline, 8-ethyl-quinoline, 1-chloroacetyl-2-hydro-3-ketone-7-acetyl-indole, 2-chloro-2',6'-diacetyl-N (methoxymethyl)acetanilide, 2-chloro-2'-acetyl-6'-ethyl-N-(methoxymethyl) acetanilide, and two hydroxylated alachlor isomers. In direct ozonation, four more byproducts were also identified including 1-chloroacetyl-2,3-dihydro-7-ethyl indole, 2-chloro-2',6'-ethyl-acetanilide, 2-chloro-2',6'-acetyl-acetanilide and 2 chloro-2'-ethyl-6'-acetyl-N-(methoxymethyl)-acetanilide. Degradation of alachlor by O(3) and O(3)/H(2)O(2) also led to the formation of low-molecular-weight byproducts including formic, acetic, propionic, monochloroacetic and oxalic acids as well as chloride ion (only detected in O(3)/H(2)O(2)). Nitrite and nitrate formation was negligible. Alachlor degradation occurred via oxidation of the arylethyl group, N-dealkylation, cyclization and cleavage of benzene ring. After O(3) or O(3)/H(2)O(2) treatment, the toxicity of alachlor solution examined by the Daphnia magna bioassay was slightly reduced. PMID- 20022077 TI - Control of hydrocarbon content of a reforming gas by using a hydrogenation catalyst. AB - To control of hydrocarbon content in waste pyrolysis-gasification and reforming processes, the use of a hydrogenation catalyst was examined in a test system with a model gas. To reduce the concentration of benzene in the reforming gas, benzene was hydrogenated with a nickel catalyst. The catalyst is usually used to convert gas-phase unsaturated hydrocarbons to saturated hydrocarbons, and the benzene was converted to cyclohexane at a temperature range of about 130 to 180 degrees C in the presence of steam. However, the conversion to methane occurred at about 250 to 300 degrees C. Methane seems to be a useful conversion compound because it does not cohere as a light tar. Sometimes the reforming gas needs to be cooled for use as generator fuel. In this case, it is possible to avoid the tar cohesion if the benzene in the gas is converted to methane at about 300 degrees C after the reforming. Reduction of the efficiency of conversion to methane was not observed over a 60h reaction period. The lower hydrocarbons (ethylene, ethane, and propylene) were also converted to methane at about 300 degrees C. Conversion of benzene was also possible when other hydrocarbons were present at high concentrations. PMID- 20022078 TI - Estimation of suspended sediment concentrations using Terra MODIS: an example from the Lower Yangtze River, China. AB - Traditional measurements of suspended sediment concentrations (SSC) through in situ sampling in rivers are expensive and time-consuming to perform. Thus, these methods cannot provide continuous SSC records. Although remote sensing has been used for SSC estimation, little research has been undertaken on inland rivers, especially for highly turbid rivers like the Yangtze. Previous studies have proposed Landsat TM/ETM+ Band 4 as a spectral SSC indicator for the Yangtze, but its limitation on temporal resolution is insufficient for the study of dynamic changes of sediment. This paper presents a method of estimating SSC of the Yangtze at Jiujiang using time-series satellite data of high temporal resolution Terra MODIS. It was found that differences in water reflectance between Band 2 and Band 5 could provide relatively accurate SSC estimation even when in-situ atmospheric conditions were unknown. After cross-validation, mean absolute relative error (ARE) and relative root mean square error (RRMSE) were relatively low (i.e., 25.5% and 36.5%, respectively). This empirical relationship was successfully applied to the estimation of SSC at Datong in the Lower Yangtze River, although the SSC values were generally underestimated. This study suggests that Terra MODIS could be used to estimate SSC in large turbid rivers, although some influencing factors require further study to improve the accuracy of SSC estimation. PMID- 20022079 TI - Current use pesticides in Arctic media; 2000-2007. AB - This review will summarize the levels of selected current use pesticides (CUPs) that have been identified and reported in Arctic media (i.e. air, water, sediment, and biota) since the year 2000. Almost all of the 10 CUPs (chlorothalonil, chlorpyrifos, dacthal, diazinon, dicofol, lindane, methoxychlor, pentachloronitrobenzene (PCNB), pentachlorophenol, and trifluralin) examined in the review currently are, or have been, high production volume chemicals i.e. >1M lbs/y in USA or >1000 t/y globally. Characteristic travel distances for the 10 chemicals range from 55 km (methoxychlor) to 12,100 km (PCNB). Surveys and long term monitoring studies have demonstrated the presence of 9 of the 10 CUPs included in this review in the Arctic environment. Only dicofol has not been reported. The presence of these chemicals has mainly been reported in high volume air samples and in snow from Arctic ice caps and lake catchments. There are many other CUPs registered for use which have not been determined in Arctic environments. The discovery of the CUPs currently measured in the Arctic has been mainly serendipitous, a result of analyzing some samples using the same suite of analytes as used for studies in mid-latitude locations. A more systematic approach is needed to assess whether other CUPs might be accumulating in the arctic and ultimately to assess whether their presence has any significance biologically or results in risks for human consumers. PMID- 20022080 TI - Characterization of Pu concentration and its isotopic composition in a reference fallout material. AB - Because there is no reference material for fallout plutonium isotope monitoring, preparation of such a material is necessary for quality control of fallout radionuclides analysis for atmospheric environmental studies. In this work, we report the characterization of Pu activity and its isotopic composition in a reference fallout material prepared by the Meteorological Research Institute (MRI), Japan. This material was prepared from samples collected at 14 stations throughout Japan in 1963-1979, with reference values of (137)Cs, (90)Sr and (239)(+)(240)Pu activities. We analyzed the activities of (239)(+)(240)Pu and (241)Pu, and the atom ratios of (240)Pu/(239)Pu and (241)Pu/(239)Pu using an isotope dilution sector-field inductively coupled plasma mass spectrometry (SF ICP-MS). The (239)(+)(240)Pu activities in this fallout material using acid leaching and total digestion were 6.56+/-0.20 mBq/g and 6.79+/-0.16 mBq/g, respectively. Atom ratios of (240)Pu/(239)Pu were 0.1915+/-0.0030 and 0.1922+/ 0.0044, respectively. Both (240)Pu/(239)Pu and (241)Pu/(239)Pu atom ratios were slightly higher than those of global fallout, which could be attributed to the deposition of fallout radionuclides resulting from the Chinese nuclear weapons tests conducted in the 1970s. The dominant host phases of (239)(+)(240)Pu were found to be organic matter-sulfides (70%) with a relative high (240)Pu/(239)Pu atom ratio, and Fe-Mn oxides (19%) using a sequential extraction method. PMID- 20022081 TI - Factor XII: what does it contribute to our understanding of the physiology and pathophysiology of hemostasis & thrombosis. AB - Factor XII (FXII) is a coagulation protein that is essential for surface activated blood coagulation tests but whose deficiency is not associated with bleeding. For over forty years, investigators in hemostasis have not considered FXII important because its deficiency is not associated with bleeding. It is because there is a dichotomy between abnormal laboratory assay findings due to FXII deficiency and clinical hemostasis that investigators sought explanations for physiologic hemostasis independent of FXII. FXII is a multidomain protein that contains two fibronectin binding consensual sequences, two epidermal growth factor regions, a kringle region, a proline-rich domain, and a catalytic domain that when proteolyzed turns into a plasma serine protease. Recent investigations with FXII deleted mice that are protected from thrombosis indicate that it contributes to the extent of developing thrombus in the intravascular compartment. These findings suggest that it has a role in thrombus formation without influencing hemostasis. Last, FXII has been newly appreciated to be a growth factor that may influence tissue injury repair and angiogenesis. These combined studies suggest that FXII may become a pharmacologic target to reduce arterial thrombosis risk and promote cell repair after injury, without influencing hemostasis. PMID- 20022082 TI - Demonstration of substances of innate immunity in the esophageal epithelium of domesticated mammals: Part II--Defence mechanisms, including species comparison. AB - The second part of our study deals with a comparative evaluation and discussion of the immunohistochemical results that were obtained. The cryoscanning electron microscopy (cryoSEM) observations confirmed a monolayer colonization of the esophageal surface with bacteria and fungi (yeasts); the latter in particular was prominent in the ruminant species studied. We demonstrated the existence of several innate immune parameters, including pathogen recognition receptors (PRRs), such as Toll-like receptor 2, which was primarily expressed in the stratum basale; however, the presence beta-glucan receptors remained inconclusive. Furthermore, the group of cationic antimicrobial peptides (CAPs) was shown, comprising beta-defensins 2 and 3 and cathelicidin. The less keratinized esophageal epithelium of the carnivorous cat was protected by high amounts of CAPs; whereas the more strongly keratinized epithelium of the herbivorous and omnivorous species with its characteristic layer structure exhibited clearly weaker reactions. Moreover, lysozyme could distinctly be demonstrated in the cells of the esophageal epithelium. It can be concluded that a first line of defence mechanisms of the innate immune system contributes to maintaining a microbial homeostasis on the surface of the esophageal epithelium of domesticated mammals. The results are discussed in comparison to findings from studies on the human esophagus. PMID- 20022083 TI - Auto amputation of penis due to advanced penile carcinoma. AB - A middle-aged man presented with progressively enlarging ulcer on the tip of penis over a year's duration and sloughing of whole penis subsequently. Examination revealed complete loss of penis and purulent discharge at its base. Biopsy of the base of the penis confirmed penile carcinoma. Imaging studies revealed extensive metastatic lesions. Palliative care was given, but he succumbed later. In this era of advanced medical care, symptoms of penile cancer are still being ignored and have led to autoamputation of penis. PMID- 20022084 TI - Comparison of the UCLA Integrated Staging System and the Leibovich score in survival prediction for patients with nonmetastatic clear cell renal cell carcinoma. AB - OBJECTIVES: To directly compare the models-the UCLA-Integrated Scoring System (UISS) and the Leibovich models-using various survival endpoints. Several Phase III trials of adjuvant therapy in renal cell carcinoma (RCC) have been initiated after advances in targeted therapy. To select patients at high risk of relapse and mortality, 2 aforementioned prognostic models have been incorporated into these trials. These models have not been compared previously. METHODS: A retrospective study of 355 patients with unilateral nonmetastatic clear cell RCC undergoing nephrectomy between 1990 and 2006 at the Singapore General Hospital was undertaken. Performance of the UISS and the Leibovich models, as well as corresponding trial inclusion criteria, was directly compared using log likelihood statistics. Adequacy and concordance indices were also calculated. Study endpoints tested were overall survival (OS), cancer-specific survival (CSS), and disease-free survival (DFS). RESULTS: Likelihood ratio testing demonstrated a significant benefit in prediction when adding the Leibovich model to the UISS model in all outcomes tested, with no benefit using the converse approach (OS: P=.002 vs P=.27; CSS: P=.0001 vs P=.57; DFS: P=<0.0001 vs P=.30). Benefit was seen primarily in disease-free survival when adding the Leibovich trial criteria to UISS trial criteria, with no benefit using the converse approach (OS: P=.16 vs P=.27; CSS: P=.17 vs P=.11; DFS: P=.01 vs P=.26). CONCLUSIONS: Both the Leibovich model and trial criteria are superior to the UISS model and trial criteria, respectively, in estimating survival outcomes in patients with nonmetastatic clear cell RCC after nephrectomy. PMID- 20022085 TI - Surgery-related complications in 1253 robot-assisted and 485 open retropubic radical prostatectomies at the Karolinska University Hospital, Sweden. AB - OBJECTIVES: To quantify complications to surgery in patients treated with robot assisted radical prostatectomy (RARP) and open retropubic radical prostatectomy (RRP) at our institution. Radical prostatectomy is associated with specific complications that can affect outcome results in patients. METHODS: Between January 2002 and August 2007, a series of 1738 consecutive patients underwent RARP (n = 1253) or RRP (n = 485) for clinically localized prostate cancer. Surgery-related complications were assessed using a prospective hospital-based complication registry. The baseline characteristics of all patients were documented preoperatively. RESULTS: Overall, 170 patients required blood transfusions (9.7%), 112 patients (23%) in the RRP group compared with 58 patients (4.8%) in the RARP group. Infectious complications occurred in 44 RRP patients (9%) compared with 18 (1%) in the RARP group. Bladder neck contracture was treated in 22 (4.5%) patients who had undergone RRP compared with 3 (0.2%) in the RARP group. Clavien grade IIIb-V complications were more common in RRP patients (n = 63; 12.9%) than in RARP patients (n = 46; 3.7%). CONCLUSIONS: The introduction of RARP at our institution has resulted in decreased number of patients with Clavien grade IIIb-V complications, such as bladder neck contractures, a decrease in the number of patients who require blood transfusions, and decreased numbers of patients with postoperative wound infections. PMID- 20022086 TI - Effects of combined use of trospium chloride and melatonin on in vitro contractility of rat urinary bladder. AB - OBJECTIVES: To examine the effects of combined use of trospium chloride and melatonin on in vitro contractility of rat urinary bladder. METHODS: Isolated bladder strips from 20 male Wistar rats were examined in an organ bath. Contractions were evoked by acetylcholine (ACh). Initially, effective concentrations of trospium and melatonin to inhibit the contractions were determined. We subsequently tested the effects of combined use of melatonin and trospium on agonist-induced contractions. RESULTS: Exposure of bladder strips to 10 microM ACh significantly increased the contractions. After cumulative administration of increasing concentrations of trospium (1, 3, and 5 microM), agonist-induced contractions decreased significantly (P <.05). Similarly, the mean peak amplitude of contractions evoked by ACh was significantly inhibited by melatonin in a concentration-dependent manner (100, 200, and 300 microM) (P <.05). Further evaluation of the effects of combined use of trospium and melatonin on ACh-induced contractions of bladder strips revealed that one-tenth of initially applied dose of trospium (500 nM) in addition to the lowest inhibitory dose of melatonin (100 microM) significantly lowered both the peak amplitude and area under the contractility curve of contractions (P <.05). CONCLUSIONS: Our results showed that combined use of low dose trospium and melatonin had strong in vitro inhibitory capability on agonist-induced contractions of rat bladder strips. The present findings may offer an insight into lowering the dose of an antimuscarinic by combining it with an antioxidant and endogenous hormone, melatonin. PMID- 20022087 TI - Intravesical instillation of hyaluronic acid prolonged the effect of bladder hydrodistention in patients with severe interstitial cystitis. AB - OBJECTIVES: To evaluate the efficacy of intravesical instillation of hyaluronic acid (HA) after hydrodistention for the treatment of patients with interstitial cystitis (IC) having small bladder capacity. METHODS: A total of 47 patients with IC (aged 27-76 years) whose functional bladder capacity was less than 200 mL received bladder hydrodistention. Thereafter, 20 patients received intravesical instillation of 40 mg HA weekly in the first month and then monthly in the following 2 months. Sixteen patients received intravesical heparin instead and 11 patients received hydrodistention alone as the control. Mean voids per day, visual analog scale for pain, and functional bladder capacity were measured before hydrodistention and 3 and 6 months after hydrodistention in all 3 groups and 9 months after hydrodistention in HA and heparin groups. RESULTS: Two patients in the HA group and 1 in the heparin group failed to complete the treatment. Three months after hydrodistention, there was no improvement in the control group. Six and 9 months after hydrodistention, rate of improvement was significantly higher in the HA group than in the heparin group (77.8% vs 33.3%, P < .05; 50% vs 20%, P < .05). At 9 months, heparin treatment did not show any improvement. Improvement in voids per day (-1.8 +/- 2.5, P < .01), visual analog scale (-0.9 +/- 1.1, P < .01), and bladder capacity (16 +/- 18 mL, P < .01) was still significant in the HA group. CONCLUSIONS: Intravesical instillation of HA may obviously prolong the effect of bladder hydrodistention in patients with severe IC. Its effect was better than heparin. PMID- 20022088 TI - Persistent Mullerian duct syndrome with torsion of a transverse testicular ectopia: first reported case. AB - Persistent Mullerian duct syndrome with transverse testicular ectopia is a rare pathologic association. A 7-month-old boy was admitted to the emergency department with a right testicular swelling. Physical examination revealed a tender right testis, the upper limits of which could not be palpated. The left testis was impalpable. An ultrasound revealed a normal right testis with an evident blood flow, overcome by a similar mass but avascular. At surgery, the right hernial sac contained both testes with an infantile uterus and fallopian tubes. The left testis was viable after manipulation. Proximal-salpingectomy with corporeal hysterectomy was performed to allow bilateral scrotal orchiopexy. PMID- 20022089 TI - Safety and efficacy of percutaneous nephrolithotomy in infants, preschool age, and older children with different sizes of instruments. AB - OBJECTIVES: To compare the morbidity and success rates among different age groups of children undergoing percutaneous nephrolithotomy (PCNL) using adult- or pediatric-sized devices. PCNL for renal stones in children may present problems because of small size, mobility of the pediatric kidney, and the small size of the collecting system. METHODS: Patients were categorized into 2 age groups: those < or =7 years old at the time of PCNL (group 1, n = 17 [38.6%]), and those 8-16 years old (group 2, n = 27 [61.4%]). Group 2 children were further divided into subgroups according to the use of pediatric- (group 2a, n = 12 [27.3%]) or adult-sized devices (group 2 b, n = 15 [34.1%]). RESULTS: Mean patient age was 4.1, 11.7, and 13.2 years in groups 1, 2a, and 2b, respectively. Fluoroscopy time, time to access the collecting system, operative time, and average postoperative hospital stay did not differ between the groups. However, hemoglobin decrease, bleeding during surgery, and blood transfusion rate was higher in group 2b. Stones were completely cleared in 82.4%, 83.3%, and 81.3% patients, and these percentages increased to 94.1%, 91.7%, and 93.7% with adjunctive shock wave lithotripsy and ureterorenoscopy in groups 1, 2a, and 2b, respectively. CONCLUSIONS: Endourologic intervention in children usually requires instruments specific for preschool age; however, in older children with dilated collecting system, the use of adult instruments and techniques may achieve equal results. PMID- 20022090 TI - Pelvic cake kidney drained by a single ureter associated with unicornuate uterus. AB - The pelvic cake kidney in which there is complete fusion of both kidneys into a single renal parenchymal mass has been described as the rarest of renal fusion anomalies. Pelvic cake kidney most commonly drains via 2 separate ureters, with fewer than 10 reports demonstrating a cake kidney drained by a single ureter. We present the first ever reported case of a pelvic cake kidney drained by a single ureter in association with a unicornuate uterus. These findings were identified in a young female patient who underwent pelvic magnetic resonance imaging for pelvic pain. PMID- 20022091 TI - Adenocarcinoma versus urothelial carcinoma of the urinary bladder: comparison between pathologic stage at radical cystectomy and cancer-specific mortality. AB - OBJECTIVES: To compare stage at radical cystectomy (RC) and cancer-specific mortality (CSM) after RC between non-urachal adenocarcinoma (ADK) and urothelial carcinoma (UC) of the urinary bladder. METHODS: Within 17 Surveillance, Epidemiology and End Results registries, we identified ADK and UC patients who underwent a RC between 1988 and 2006. We examined differences in stage and grade at RC between ADK and UC patients. Kaplan-Meier plots depicted CSM after RC. Cox regression analyses examined CSM rates, adjusted for T and N stages, tumor grade, age, gender, race, and year of surgery. Thereafter, we relied on statistically significant variables from the multivariate Cox regression model to match ADK and UC patients. Finally, we plotted Kaplan-Meier survival curves of the matched ADK and UC patients. RESULTS: Of 306 ADK and 11 697 UC patients, 188 (61.4%) and 5538 (47.3%), respectively, showed extravesical disease (pT(3-4); P <.001) and 26.5% vs 21.7% had lymph node metastases at RC (P = .04), respectively. After adjustment for all covariates, including stage and grade, ADK was not associated with worse prognosis than UC (hazard ratio, 1.05; P = .6). Similarly, after matching, no difference in CSM was recorded between the 2 histologic subtypes (hazard ratio, 1.07; P = .5). CONCLUSIONS: ADK patients undergo RC at more advanced disease stages. However, stage- and grade-adjusted CSM is the same between ADK and UC patients. Efforts should be aimed at providing definitive treatment at earlier stages, especially in patients with ADK histologic subtype. PMID- 20022092 TI - The effect of primary cytoreduction on outcomes of patients with FIGO stage IIIC ovarian cancer stratified by the initial tumor burden in the upper abdomen cephalad to the greater omentum. AB - OBJECTIVE: Our objective was to analyze the effect of surgical outcome on progression-free survival (PFS) and overall survival (OS) of patients with advanced ovarian carcinoma stratified by the initial presence and volume of upper abdominal disease cephalad to the greater omentum (UAD) found at the time of exploration. METHODS: We evaluated all patients with FIGO stage IIIC ovarian carcinoma who underwent primary cytoreduction followed by platinum-based chemotherapy at our institution between January 1989 and December 2006. The effect of surgical outcome was investigated using a time-to-event analysis. A Cox proportional hazards model was fit using clinical, surgical, and postoperative variables. RESULTS: We identified 526 evaluable patients. Optimal versus suboptimal cytoreduction was significantly associated with improved median PFS and OS in patients with no, minimal (1 cm) UAD. On multivariate analysis, patients with bulky UAD who underwent optimal cytoreduction had a 28% decreased risk of relapse (hazard ratio, 0.72; 95% confidence interval: 0.53-0.99; P=0.04) and a 33% decreased risk of death (hazard ratio, 0.67; 95% confidence interval: 0.47-0.96; P=0.03) compared to patients who underwent suboptimal cytoreduction. CONCLUSION: The presence of large-volume disease found during surgical exploration does not preclude the benefit of optimal cytoreduction. The findings support the management strategy of maximizing surgical efforts with increasing tumor burden in patients with stage IIIC ovarian cancer. Prospective studies are needed to more precisely quantify tumor burden and accurately determine the specific impact of cytoreduction on outcome. PMID- 20022093 TI - RhoC expression level is correlated with the clinicopathological characteristics of ovarian cancer and the expression levels of ROCK-I, VEGF, and MMP9. AB - OBJECTIVE: To determine the clinicopathological significance of RhoC expression in human ovarian cancer and its effect on the expression of vascular endothelial growth factor (VEGF), Rho-associated coiled-coil-forming kinase (ROCK), and metal matrix proteinases (MMPs). METHODS: Tissue samples from normal ovaries, benign ovarian tumors, and epithelial ovarian cancer were collected. The mRNA and protein expression levels of RhoC, ROCK-I, VEGF, and MMP9 were assessed using reverse-transcriptase polymerase chain reaction (RT-PCR) and Western blot, and compared to the clinicopathological characteristics of the sample of origin using the Pearson method of correlation analysis. Small interfering RNA (siRNA) was also used to target RhoC expression in the OVCAR3 and CaOV3 ovarian cancer cell lines, after which cell invasion and migration assays were performed, and the expression of ROCK-I, VEGF, and MMP9 was evaluated. RESULTS: The expression levels of RhoC, ROCK-I, VEGF, and MMP9 mRNA and protein were significantly higher in ovarian cancer, showing a correlation with clinical stage but not histological type. RhoC expression was positively correlated with ROCK-I, VEGF, and MMP9 expression. Decreased RhoC expression in siRNA-targeted cells inhibited their ability to invade and migrate, as well as inhibiting ROCK-I, VEGF, and MMP9 expression. CONCLUSION: The expression level of RhoC is correlated to clinical stage and vascularization in ovarian cancer. PMID- 20022094 TI - A nomogram for predicting overall survival of women with endometrial cancer following primary therapy: toward improving individualized cancer care. AB - OBJECTIVES: Traditionally we have relied mainly on final FIGO stage to estimate overall oncologic outcome in endometrial cancer patients. However, it is well known that other patient factors may play equally important roles in outcome. Our objective was to develop a clinically useful nomogram in the hope of providing a more individualized and accurate estimation of overall survival (OS) following primary therapy. METHODS: Using a prospectively maintained endometrial cancer database, 1735 patients treated between 1993 and 2008 were analyzed. Characteristics known to predict OS were collected. For each patient, points were assigned to each of these 5 variables. A total score was calculated. The association between each predictor and the outcome was assessed by multivariable modeling. The corresponding 3-year OS probabilities were then determined from the nomogram. RESULTS: The median age was 62 years (range, 25-96). Final grade included: G1 (471), G2 (622), G3 (634), and missing (8). Stage included: IA (501), IB (590), IC (141), IIA (36), IIB (75), IIIA (116), IIIB (6), IIIC (135), IVA (7), and IVB (128). Histology included: adenocarcinoma (1376), carcinosarcoma (100), clear cell (62), and serous (197). Median follow-up for survivors was 29.2 months (0-162.2 months). Concordance probability estimator for the nomogram is 0.746+/-0.011. CONCLUSION: We developed a nomogram based on 5 easily available clinical characteristics to predict OS with a high concordance probability. This nomogram incorporates other individualized patient variables beyond FIGO stage to more accurately predict outcome. This new tool may be useful to clinicians in assessing patient risk when deciding on follow-up strategies. PMID- 20022095 TI - Respiratory and cardiovascular effects of doxapram and theophylline for the treatment of asphyxia in neonatal calves. AB - Respiratory stimulants are widely used in asphyxic neonatal calves despite a lack of data about their effectiveness and indications of possible side effects. The effect of doxapram and theophylline on respiratory, cardiovascular, and acid-base variables was investigated in 10 healthy neonatal calves (Bos Taurus). A venous, a peripheral arterial, and a pulmonary arterial catheter were placed, and central venous, pulmonary, and systemic blood pressures and cardiac output were measured using thermodilution technique. Doxapram, but not theophylline, led to an immediate increase in respiratory rate (P or=2 fields of both eyes. MAIN OUTCOME MEASURES: Any RVO, CRVO, or BRVO. RESULTS: The combined pooled data contained 68,751 individuals from 15 studies, with participants' ages ranging from 30 to 101 years. In analyses of 11 studies that assessed >or=2 fundus fields of both eyes (n=49,869), the age- and sex-standardized prevalence was 5.20 per 1000 (confidence interval [CI], 4.40 5.99) for any RVO, 4.42 per 1000 (CI, 3.65-5.19) for BRVO, and 0.80 per 1000 (CI, 0.61-0.99) for CRVO. Prevalence varied by race/ethnicity and increased with age, but did not differ by gender. The age- and sex-standardized prevalence of any RVO was 3.7 per 1000 (CI, 2.8-4.6) in whites (5 studies), 3.9 per 1000 (CI, 1.8-6.0) in blacks (1 study), 5.7 per 1000 (CI, 4.5-6.8) in Asians (6 studies), and 6.9 per 1000 (CI, 5.7-8.3) in Hispanics (3 studies). Prevalence for CRVO was lower than BRVO in all ethnic populations. On the basis of these data, an estimated 16.4 million (CI, 13.9-18.9) adults are affected by RVO, with 2.5 million (CI, 1.9-3.1) affected by CRVO and 13.9 million (CI, 11.5-16.4) affected by BRVO. Study limitations include non-uniform sampling frames in identifying study participants and in acquisition and grading of RVO data. CONCLUSIONS: Our study provides summary data on the prevalence of RVO and suggests that approximately 16 million people may have this condition. Research on preventive and treatment strategies for this sight-threatening eye disease is needed. PMID- 20022119 TI - Platelet monoamine oxidase activity in children with attention deficit/hyperactivity disorder. AB - Attention-deficit/hyperactivity disorder (ADHD) is a highly heritable developmental disorder characterized by symptoms of impulsivity, hyperactivity and/or inattention, and associated with structural and biochemical abnormalities in cortical and limbic structures innervated by dopamine, noradrenalin and serotonin. The enzyme monoamine oxidase, type B (MAO-B), is expressed in platelets, and metabolizes endogenous amines. Its activity has been proposed to represent a peripheral marker of various traits and forms of psychopathology. This study evaluated platelet MAO activity with a spectrofluorimetric method in 72 boys and 12 girls with predominantly hyperactive, predominantly inattentive, and combined subtype of ADHD (DSM-IV criteria), and in 64 control children. The results showed significantly lower platelet MAO activity in children with hyperactive, inattentive, and combined subtype of ADHD than in control children. There was no significant association between platelet MAO activity and gender or age. The limitation of the study was in the small sample of girls with ADHD (N=12), and in the determination of only one peripheral marker. In line with hypotheses of lower platelet MAO activity in different types of psychopathology, children with different subtypes of ADHD had significantly lower platelet MAO-B activity than control children. PMID- 20022120 TI - Effectiveness of a psychosocial intervention for relapse prevention in patients with schizophrenia receiving risperidone via long-acting injection. AB - Schizophrenia is a chronic disorder that is usually characterized by relapses alternating with periods of full or partial remission. We examined whether combined therapy with a psychosocial intervention for relapse prevention (PIRP) and risperidone administered by long-acting injection (RLAI) would be more effective in reducing relapses than RLAI with treatment-as-usual (TAU) among outpatients with schizophrenia. We conducted a prospective, controlled study over 2 years in 46 patients with schizophrenia receiving RLAI, of which 21 and 25 patients were assigned to the PIRP and TAU control groups, respectively. The 1- and 2-year relapse rates were lower and medication compliance was higher in the PIRP group than in the TAU group. Cox proportional analysis revealed that time from baseline to relapse was associated with RLAI discontinuation. These results indicate that PIRP can be effective in maintaining medication compliance, and that discontinuation of long-acting atypical antipsychotics might be predictive of the next relapse. However, these results need to be replicated in studies with larger samples. PMID- 20022122 TI - Sex-based differences in answering strategy and the influence of cross-sex hormones. AB - We investigated whether sex differences in answering strategy occur in normal controls (C). Furthermore, it was tested whether these sex differences were subject to change over time, and whether they were associated with hormonal treatment at time points 2 and 3 in patients with Gender Identity Disorder (GID). Two subtests measuring arithmetic ability were used: arithmetic aptitude (AA) and arithmetic operations (AO). Both the controls (n = 29) and GID patients (n = 33) were tested at baseline (T1), three months (T2) and 12 months (T3) after the start of hormonal treatment in the GID group. A repeated measures analysis of variance showed no differences between C males and females, for T1 and T2. At T3, C males guessed more than C females. At baseline, GID males and C males left an equal number of items unanswered. However, when being retested, C males left fewer items unanswered than GID males. No difference was found between C females and GID females at any time point. Our results suggest that healthy adult males become more confident when they are retested, and seem to adjust their answering strategy accordingly. Moreover, hormonal treatment of healthy adult GID patients born male is associated with a lack of adjustment in answering strategy. PMID- 20022121 TI - Recognition of facial affect in girls with conduct disorder. AB - Impaired recognition of facial affect has been reported in youths and adults with antisocial behavior. However, few of these studies have examined subjects with the psychiatric disorders associated with antisocial behavior, and there are virtually no data on females. Our goal was to determine if facial affect recognition was impaired in adolescent girls with conduct disorder (CD). Performance on the Ekman Pictures of Facial Affect (POFA) task was compared in 35 girls with CD (mean age of 17.9 years+/-0.95; 38.9% African-American) and 30 girls who had no lifetime history of psychiatric disorder (mean age of 17.6 years+/-0.77; 30% African-American). Forty-five slides representing the six emotions in the POFA were presented one at a time; stimulus duration was 5s. Multivariate analyses indicated that CD vs. control status was not significantly associated with the total number of correct answers nor the number of correct answers for any specific emotion. Effect sizes were all considered small. Within CD analyses did not demonstrate a significant effect for aggressive antisocial behavior on facial affect recognition. Our findings suggest that girls with CD are not impaired in facial affect recognition. However, we did find that girls with a history of trauma/neglect made a greater number of errors in recognizing fearful faces. Explanations for these findings are discussed and implications for future research presented. PMID- 20022123 TI - Targeting of Escherichia coli F4 fimbriae to Fcgamma receptors enhances the maturation of porcine dendritic cells. AB - F4(+) enterotoxigenic Escherichia coli (ETEC) infections are an important cause of postweaning diarrhoea in piglets and an oral immunization of piglets with purified F4 fimbriae protects them from a subsequent F4(+) ETEC infection. However, oral immunization of suckling piglets is hampered due to the immature status of their immune system. Targeting of antigens to Fcgamma receptors (FcgammaR) on human and murine dendritic cells (DC) has been shown to enhance DC maturation and both humoral and cellular immune responses. To investigate the effect of F4 fimbriae incorporated in immune complexes (F4-IC) on porcine DC, we used porcine monocytic-derived DC (MoDC) as a model system. The results in this study demonstrate that FcgammaRI, II and III mRNA is expressed by porcine MoDC. Furthermore, we show that FcgammaRII and III are expressed on the cell surface and that F4-IC are internalized by MoDC via FcgammaR. This FcgammaR ligation induced a significantly enhanced expression of Major Histocompatibility complex (MHCII) class II and the costimulatory molecules CD80/86 and CD40 by MoDC compared with immature MoDC. Furthermore, the phagocytic capacity of F4-IC stimulated MoDC was reduced as evidenced by a reduced uptake of DQ-ovalbumin and FITC-dextran. In an allogenic and autologous mixed lymphocyte reaction, these F4 IC-activated MoDC showed an improved T cell stimulatory capacity in comparison with immature MoDC. The F4-IC induced DC maturation correlated with significant higher expression levels of several pro-inflammatory cytokines such as interleukine (IL) 1beta, IL-6 and Tumor necrosis factor alpha, the chemokine IL-8 and IL-12p40 in comparison with immature MoDC. Altogether, these results clearly demonstrate that FcgammaR engagement enhances the maturation of porcine MoDC, which may suggest that antigen targeting to FcgammaR on DC could improve vaccine design against infections. PMID- 20022124 TI - Mutational analysis of the PITX2 and NKX2-5 genes in patients with idiopathic atrial fibrillation. AB - Atrial fibrillation (AF) is the most frequently encountered arrhythmia in clinical practice. In a subgroup of patients, AF is regarded as idiopathic when no signs of structural heart disease or other causes of the arrhythmia can be identified during conventional clinical work-up. Recent studies have demonstrated that AF has a substantial genetic basis in a number of cases. The entire coding sequences, including intron-exon boundaries, of the genes PITX2 and NKX2-5 were screened for genetic variants by means of initial polymerase chain reaction followed by DNA sequencing in 96 patients with idiopathic AF. Although we detected a number of variants, our candidate gene approach did not result in identification of mutations associated with AF in the coding regions of PITX2 or NKX2-5 in our well characterized AF cohort. PMID- 20022125 TI - A model system for evaluating surface disinfection in dairy factory environments. AB - A model system was developed for evaluating the efficacy of disinfectants for inactivating bacteria present in biofilms on surfaces within dairy factory environments. Mixed culture biofilms of six dairy factory isolates (pseudomonads, coliforms and presumptive staphylococci) were generated on factory floor tiles and subjected to up to three fouling and cleaning (FC) cycles. Disinfectants (hypochlorite, peroxyacetic acid-, acid anionic- and quaternary ammonium compound (QUAT)-based products) were applied after cleaning at the supplier's recommended concentrations. Bacterial groups were enumerated using selective agar media. All disinfectants significantly (p<0.05) reduced surface counts of the three bacterial groups. The peroxyacetic acid-based product applied at 3.0% (v/v) achieved the greatest overall reductions in counts. In some cases, disinfectants were more biocidal towards particular bacterial groups. For example, hypochlorite demonstrated greater biocidal activity towards coliforms and staphylococci while the QUAT- and acid anionic-based disinfectants demonstrated greater biocidal activity towards staphylococci. The selective biocidal activity of hypochlorite and the acid anionic-based disinfectant was maintained over at least three FC cycles. The results demonstrate that disinfectants can alter the composition of the surface microflora and can be used to selectively reduce or eliminate particular bacterial groups from production environments. The model system was adapted to assess biofilm development and inactivation on a range of floor materials used in dairy factory environments. Clear differences were observed between surface materials at the end of the inoculation stage and following one FC cycle. For example, after one FC cycle, the highest bacterial counts were obtained for extruded floor tiles and the lowest for stainless steel and the polyurethane surface coating. The model system is versatile and there is scope to employ it to investigate a range of factors influencing the efficacy of cleaning and disinfection procedures towards biofilms. In addition, other microorganisms, such as pathogenic bacteria, may be added to the model biofilm and the influence of disinfectants on these investigated. PMID- 20022126 TI - Trends in lower extremity amputations in people with and without diabetes in England, 1996-2005. AB - AIMS: To examine trends in non-traumatic lower extremity amputations over a 10 year-period in people with and without diabetes (DM) in England. METHODS: All individuals admitted to NHS hospitals for non-traumatic amputations between 1996 and 2005 in England were identified using hospital activity data. Postoperative and 1-year mortality were examined between 2000 and 2004. RESULTS: There was a reduction in minor and major amputations during the study period. The number of type 1 DM- and non-DM-related minor amputations decreased by 11.4% and 32.4%, respectively, while the number of type 2 DM-related minor amputations almost doubled. The incidence of type 1- and non-DM-related minor amputations decreased from 1.5 to 1.2 and from 8.1 to 5.1/100,000 population, respectively, while type 2 DM-related amputations increased from 2.4 to 4.1/100,000 population. The number of type 1- and non-DM-related major amputations declined by 41% and 22%, respectively, whereas type 2 DM-related amputations increased by 43%. The incidence of type 2 DM-related amputations increased from 2.0 to 2.7/100,000 population. Overall perioperative and 1-year mortality did not significantly change between 2000 and 2004. CONCLUSIONS: While several factors may explain the increase in type 2 DM-related LEAs, these findings highlight the importance of diabetes prevention strategies and controlling risk factors for LEAs in people with diabetes. PMID- 20022127 TI - Time lag characterization of two continuous glucose monitoring systems. AB - AIM: To evaluate the system time lag associated with subcutaneous interstitial glucose (IG) sensing and venous blood glucose (BG) of two continuous glucose monitoring (CGM) systems, the SEVEN((R)) (DexCom, San Diego, CA) and the Navigator((R)) (Abbott Diabetes Care, Alameda, CA), in adults with type 1 diabetes. METHODS: Fourteen subjects wore both CGM systems concurrently during the 15-day study. Reference YSI (Yellow Springs, OH) and CGM data from the in clinic sessions, conducted on day 5, 10 and 15 of the study were evaluated. The system time lag of CGM system was estimated using various regression method related statistical estimators. RESULTS: The estimated time lags based on different statistical measures are similar within each CGM system. The time lag based on correlation coefficient criteria is estimated as 4.5+/-5min (median+/ IQR) for the SEVEN((R)), and 15+/-7min for the Navigator((R)). The ranges of these estimators of two CGM systems were different (2-6min for SEVEN((R)) and 14 15min for Navigator((R))). CONCLUSIONS: The study findings suggest that commonly accessible statistics, such as correlation statistics, offer estimates that are comparable to complicated approaches. Different time lags were observed with two CGM systems. PMID- 20022128 TI - Serum fatty acid composition and insulin resistance are independently associated with liver fat markers in elderly men. AB - AIM: To investigate the relationships of serum fatty acid (FA) composition and estimated desaturase activities with the liver fat marker alanine aminotransferase (ALT). METHODS: 546 Swedish elderly men of a population-based cohort participated in this cross-sectional study. FA composition was assessed in serum cholesterol esters to determine dietary fat quality (e.g. linoleic) and desaturation products (e.g. dihomo-gamma-linolenic acid). Desaturase indices, including stearoyl coenzymeA desaturase-1 (SCD-1), were calculated by FA product to-precursor ratios. RESULTS: In linear regression analyses adjusting for lifestyle, abdominal obesity and insulin sensitivity, the dietary biomarker linoleic acid (n-6), but not n-3 FAs, was inversely related to ALT. Desaturation products including palmitoleic, oleic, gamma-linolenic and dihomo-gamma-linolenic acids, and Delta6-desaturase and SCD-1 indices were directly related to ALT (all p<0.05). After further adjustment for factors previously linked to fatty liver (i.e. serum lipids, adiponectin concentrations), SCD-1 index (p=0.004) and insulin resistance (p<0.0001) were independent determinants of ALT activity, whereas waist circumference, triglycerides, non-esterified FA and adiponectin were not. CONCLUSION: A low dietary intake of linoleic acid and elevated SCD-1 index may contribute to higher ALT activity in elderly men, even independently of obesity and insulin resistance. PMID- 20022129 TI - NF-kappaB signalling: embracing complexity to achieve translation. AB - NF-kappaB is a dimeric transcription factor that has emerged as an important regulator of liver homeostasis and is mechanistically implicated in a variety of liver pathologies including hepatitis, steatosis, fibrosis, and hepatocellular carcinoma. The question remains as to whether NF-kappaB can really be exploited for the development of therapeutics for these pathologies in the diseased human liver. This review casts a critical eye on the experimental evidence gathered to date and in particular questions the rationale for the current focus on components of the upstream IKK complex as therapeutic targets. We make the argument that translation of NF-kappaB biology to new therapies is more likely to emerge from a re-focus of basic research back to the NF-kappaB/Rel subunit functions and the complexities of their post-translational modifications and context-dependent co-regulator interactions. PMID- 20022130 TI - Liver stiffness is directly influenced by central venous pressure. AB - BACKGROUND & AIMS: Liver stiffness (LS) as measured by transient elastography [Fibroscan] offers a novel non-invasive approach to assess liver cirrhosis. Since Fibroscan seems to be unreliable in patients with congestive heart failure, it remains to be determined whether hemodynamic changes affect LS irrespective of fibrosis. METHODS & RESULTS: Using landrace pigs, we studied the direct relationship between the central venous pressure and LS measured by Fibroscan. Clamping of the inferior caval vein increased LS from 3.1 to 27.8kPa while reopening reversed LS within 5min to almost normal values of 5.1kPa. We then studied LS as a function of venous pressure in the isolated pig liver by clamping the upper and lower caval, portal vein and hepatic artery. The stepwise increase of intravenous pressure to 36cm of water column (3.5kPa) linearly and reversibly increased LS to the upper detection limit of 75kPa. We finally measured LS in 10 patients with decompensated congestive heart failure before and after recompensation. Initial LS was elevated in all patients, in 8 of them to a degree that suggested liver cirrhosis (median 40.7kPa). Upon recompensation with a median weight loss of 3.0kg, LS decreased in all 10 patients down to a median LS of 17.8kPa. Inflammation could not account for increased LS since initial liver enzyme counts were only slightly elevated and did not change significantly. CONCLUSION: LS is a direct function of central venous pressure which should be considered when assessing the degree of fibrosis. PMID- 20022131 TI - Policy and practice, lost in transition: Reasons for high drop-out from pre antiretroviral care in a resource-poor setting of eastern Uganda. AB - OBJECTIVE: This study explores reasons for drop-out from pre-ARV care in a resource-poor setting where premature death is a common consequence of delayed ARV initiation. METHODS: In Iganga, Uganda, we conducted key informant interviews with staff at the pre-ARV clinic, focus group discussions with persons who looked after people living with HIV (PLWH) and in-depth interviews with PLWH half of whom had dropped out of pre-ARV care. Content data analysis was done to identify recurrent themes. RESULTS: Reasons cited for dropping out of pre-ARV care include: inadequate post-test counseling due to staff work overload, competition from the holistic and less stigmatizing traditional/spiritual healers. Others were transportation costs, long waiting time lack of incentives to seek pre-ARV care by healthy looking PLWH and gender inequalities. CONCLUSIONS: Pre-ARV adherence counseling should be improved through recruitment of counselors or multi-skilling in counseling skills for the available staff to reduce on the work load. Traditional/ spiritual healers should be integrated and supervised to offer pre-ARV care. Door step supply of cotrimoxazole using agents could reduce transport costs, waiting time and increase access to pre-ARV. Women should be sensitized on comprehensive HIV care through the local media and local leaders to address gender inequalities. PMID- 20022132 TI - Reliable in their failure: an analysis of healthcare reform policies in public systems. AB - In this paper, we analyze recommendations of past governmental commissions and their implementation in Quebec as a case to discuss the obstacles that litter the road to healthcare system reform. Our analysis shows that the obstacles to tackling the healthcare system's main problems may have less to do with programmatic (what to do) than with political and governance (how to do it) questions. We then draw on neo-institutional theory to discuss the causes and effects of this situation. PMID- 20022133 TI - Web-based interactive 2D/3D medical image processing and visualization software. AB - There are many medical image processing software tools available for research and diagnosis purposes. However, most of these tools are available only as local applications. This limits the accessibility of the software to a specific machine, and thus the data and processing power of that application are not available to other workstations. Further, there are operating system and processing power limitations which prevent such applications from running on every type of workstation. By developing web-based tools, it is possible for users to access the medical image processing functionalities wherever the internet is available. In this paper, we introduce a pure web-based, interactive, extendable, 2D and 3D medical image processing and visualization application that requires no client installation. Our software uses a four-layered design consisting of an algorithm layer, web-user-interface layer, server communication layer, and wrapper layer. To compete with extendibility of the current local medical image processing software, each layer is highly independent of other layers. A wide range of medical image preprocessing, registration, and segmentation methods are implemented using open source libraries. Desktop-like user interaction is provided by using AJAX technology in the web-user-interface. For the visualization functionality of the software, the VRML standard is used to provide 3D features over the web. Integration of these technologies has allowed implementation of our purely web-based software with high functionality without requiring powerful computational resources in the client side. The user-interface is designed such that the users can select appropriate parameters for practical research and clinical studies. PMID- 20022134 TI - Pulmonary amyloidosis mimicking multiple metastatic lesions on F-18 FDG PET/CT. AB - We report a case of amyloid pulmonary nodules with positive FDG uptake that mimic multiple lung metastases. A 54-year-old female patient was referred for the evaluation of multiple lung nodules. A PET/CT scan revealed mild FDG uptake in various sized pulmonary nodules. Resected nodules contained amorphous eosinophilic proteineous material with focal calcification, consistent with amyloidosis. Pulmonary amyloidosis should be added to the differential diagnosis for cases of multiple pulmonary nodules that show positive FDG uptake. PMID- 20022135 TI - Increased IL-17-producing cells correlate with poor survival and lymphangiogenesis in NSCLC patients. AB - The presence of IL-17-positive cells is observed in a variety of inflammatory associated cancers and IL-17 has been found to be involved in angiogenesis. The aim of this study is to determine the prognostic significance of IL-17 in NSCLC patients and to examine the correlation between IL-17 expression and lymphatic vessel density in NSCLC tissues. The expression of IL-17 was measured by immunohistochemistry in 52 paraffin-embedded tissues with non-small cell lung cancer. The chi(2) test was used to analyze the correlation between IL-17 expression and clinical parameters and lymphatic vessel density (LVD). The Kaplan Meier method, univariate and multivariate regression analysis was used to analyze the correlation between IL-17 expression and overall survival and disease-free survival. High expression of IL-17 was observed in 25 of 52 lung cancer patients and was associated with smoking status, TNM stage, LVD, overall survival and disease-free survival. Univariate and multivariate analysis showed that IL-17 was an independent prognostic factor for overall survival and disease-free survival. Our results indicate that IL-17 may play a role in the metastasis of lung cancer by promoting lymphangiogenesis. IL-17 expression is an independent prognostic factor in both overall and disease-free survival in NSCLC. PMID- 20022136 TI - Monoclonal antibody S60-4-14 reveals diagnostic potential in the identification of Pseudomonas aeruginosa in lung tissues of cystic fibrosis patients. AB - The lipopolysaccharide (LPS) of Pseudomonas aeruginosa has been identified to contain an inner-core structure expressing a Pseudomonas-specific epitope. This target structure is characterized by a highly phosphorylated and 7-O-carbamoyl-l glycero-alpha-d-manno-heptopyranose (CmHep) and was found to be present in all human-pathogenic Pseudomonas species of the Palleroni (RNA)-classification I scheme. We raised and selected the monoclonal antibody S60-4-14 (mAb S60-4-14, subtype IgG1) from mice immunized with heat-killed Pseudomonas bacteria. The epitope of this mAb was found to reside in the inner-core structure of P. aeruginosa and, hence, successfully evaluated for the immunohistochemical detection of P. aeruginosa in formalin- or HOPE-fixed (Hepes-glutamic acid buffer mediated organic solvent protection effect) and paraffin-embedded human lung tissue slices. Lung specimens, mainly from explanted lungs of cystic fibrosis (CF) patients, as well as P. aeruginosa isolates from patients suffering from CF and patients with extrapulmonar Pseudomonas infections were investigated by PCR, immunohistochemistry, and Western blot analysis with mAb S60-4-14. The results revealed an unequivocal coincidence of PCR and immunohistochemistry. Together with the Western blot results mAb S60-4-14 displays a potential diagnostic tool for the specific identification of P. aeruginosa in infected lungs of CF. PMID- 20022137 TI - Overexpression of Arabidopsis CBF1 gene in transgenic tobacco alleviates photoinhibition of PSII and PSI during chilling stress under low irradiance. AB - A known arabidopsis cDNA clone, the CRT/DRE binding factor 1 (CBF1), was isolated and introduced into tobacco plants. It has been reported that CBF1 is one member of CBF gene family related to low temperature and enhancing low temperature tolerance of plants. In the present work, the transcripts could be detected in the transgenic lines. The photochemical efficiency of PSII (F(v)/F(m)) and the photo-oxidizable P700 in the transgenic lines overexpressing CBF1 were higher than that in the wild type plants during the chilling stress under low irradiance. Similarly, the higher NPQ, higher qf, lower Phi(NF), higher activity of SOD, and lower content of MDA were also detected in the transgenic tobacco lines. Additionally, higher expression levels of Nicotiana tabacum copper/zinc superoxide dismutase (Cu/Zn SOD) were also detected in the transgenic lines. These results suggest that CBF1 protein plays an important role in protection of PSII and PSI during the chilling stress under low irradiance. PMID- 20022138 TI - Copper exposure interferes with the regulation of the uptake, distribution and metabolism of sulfate in Chinese cabbage. AB - Exposure of Chinese cabbage (Brassica pekinensis) to enhanced Cu(2+) concentrations (1-10 microM) resulted in leaf chlorosis, a loss of photosynthetic capacity and lower biomass production at > or = 5 microM. The decrease in pigment content was likely not the consequence of degradation, but due to hindered chloroplast development upon Cu exposure. The Cu content of the root increased with the Cu(2+) concentration (up to 40-fold), though only a minor proportion (4%) was transferred to the shoot. The nitrate uptake by the root was substantially reduced at > or = 5 microM Cu(2+). The nitrogen content of the root was affected little at lower Cu(2+) levels, whereas that in the shoot was decreased at > or = 5 microM Cu(2+). Cu affected the uptake, distribution and metabolism of sulfate in Chinese cabbage. The total sulfur content of the shoot was increased at > or = 2 microM Cu(2+), which could be attributed mainly to an increase in sulfate content. Moreover, there was a strong increase in water soluble non-protein thiol content in the root and, to a lesser extent, in the shoot at > or = 1 microM, which could only partially be ascribed to a Cu-induced enhancement of the phytochelatin content. The nitrate uptake by the root was substantially reduced at > or = 5 microM Cu(2+), coinciding with a decrease in biomass production. However, the activity of the sulfate transporters in the root was slightly enhanced at 2 and 5 microM Cu(2+), accompanied by enhanced expression of the Group 1 high affinity transporter Sultr1;2, and the Group 4 transporters Sultr4;1 and Sultr4;2. In the shoot, there was an induction of expression of Sultr4;2 at 5 and 10 microM Cu(2+). The expression of APS reductase was affected little in the root and shoot up to 10 microM Cu(2+). The upregulation of the sulfate transporters may be due not only to greater sulfur demand at higher Cu levels, but also the consequence of interference by Cu with the signal transduction pathway regulating the expression and activity of the sulfate transporters. PMID- 20022139 TI - MCI patients' EEGs show group differences between those who progress and those who do not progress to AD. AB - The theta/gamma and alpha3/alpha2 ratio were investigated as early markers for prognosticating of progression to dementia. 76 subjects with mild cognitive impairment (MCI) underwent EEG recording, MRI scans and neuropsychological (NPS) tests. After 3 years of follow-up, three subgroups were characterized as converters to Alzheimer's disease (AD, N=18), converters to non-AD dementia (N=14) and non-converters (N=44). The theta/gamma and alpha3/alpha2 ratio, performance on cognitive tests and hippocampal volume, as evaluated at the time of initial MCI diagnosis, were studied in the three groups. As expected, MCI to AD converters had the smallest mean hippocampal volume and poorest performance on verbal learning tests, whereas MCI to non-AD converters had poorest cognitive performance in non-verbal learning tests, abstract thinking, and letter fluency. Increased theta/gamma ratio was associated with conversion to both AD and non-AD dementia; increased alpha3/alpha2 ratio was only associated with conversion to AD. Theta/gamma and alpha3/alpha2 ratio could be promising prognostic markers in MCI patients. In particular, the increase of high alpha frequency seems to be associated with conversion in AD. EEG markers allow a mean correct percentage of correct classification up to 88.3%. Future prospective studies are needed to evaluate the sensitivity and specificity of these measures for predicting an AD outcome. PMID- 20022140 TI - Clinical impact of (18)F-FDG PET in management of patients with renal cell carcinoma. AB - We performed a retrospective study to evaluate the accuracy, diagnostic validity, and clinical impact of (18)F-FDG PET in the management of recurrent and metastatic disease in patients with Renal Cell Carcinoma (RCC) from our database. (18)F-FDG PET studies were identified from 58 patients that matched our criteria for inclusion in the study. Results were confirmed with histopathological findings, clinical follow-up time (at least 12 months), and/ or conventional imaging methods (CIM). A sensitivity of 80.56%, specificity 86.36%, diagnostic accuracy 58.7%, positive predictive value 90.63%, and a negative predictive value of 73.08% were observed. The clinical impact was high in 25 cases (43%) and we found no impact in only 10 studies (17.2%). We concluded that (18)F-FDG PET was useful and had a high clinical impact in the management of recurrent and metastatic RCC. From our data, it seemed that a positive PET study was more helpful to the physician than a negative study. PMID- 20022141 TI - [Luisa, for instance]. PMID- 20022142 TI - [I was born a woman, the life of Mame: the dramatization of intergenerational inclusion]. AB - I was born a woman, the life of Mame is the dramatization a fictitious intergenerational story. However, the story is based on true events and the feelings of real immigrant women. The dramatization narrates the life of a woman, from the time her parents-who had recently emigrated from Senegal-settled in Andalusia, up to the adolescence of Mame's own daughter. The story is told with one person on stage who, through the use of three costume changes, plays three different generations of women: mother, daughter and granddaughter, with the video-recorded testimonies of other characters (a teacher and a midwife), who express their views of the host society against a background of African music and pictures of their native country. The present article describes their goals, such as communication in conferences, and includes a concise summary of the script, reflections on the process of integrating, and the evaluation made after a performance. PMID- 20022143 TI - [On the reliability of information sources and the ethics of their use]. PMID- 20022144 TI - [Impact of a health safety warning on strontium ranelate utilization]. AB - OBJECTIVE: To evaluate changes in the use of strontium ranelate following a health safety warning in November 2007 on the risks of the development of DRESS (Drug Rash with Eosinophilia and Systemic Symptoms) with the use of this medicine. METHOD: Monthly strontium ranelate DHD (daily defined doses/1,000 inhabitants/day) were calculated between 2006 and 2008 in a health area of Spain. The data were analyzed graphically and through a multiple regression model. RESULTS: Graphical representation showed a long-term rising trend throughout the study period, although growth slowed after the safety warning was issued. The multiple regression model showed that the health warning was statistically significant. CONCLUSION: The use of strontium ranelate continues to grow after the issue of the health safety warning, although at a slower pace than previously. PMID- 20022145 TI - [Quality of data used to calculate reproductive and perinatal health indicators in native and migrant populations]. AB - OBJECTIVE: To assess the quality of data on births in the Natural Population Movement (NPM) and congenital metabolic disorders registers with regard to calculation of reproductive and perinatal health indicators. METHODS: The following comparisons between registers were made: (1) the total number of births to mothers living in Catalonia and Valencia from 2005 to 2006, (2) the percentage of missing data on the mother's geographical origin, (3) the percentage of missing data on the mother's age and the infant's birthweight and gestational age according to maternal origin. RESULTS: The congenital metabolic disorders registers exhaustively collected the total number of births gathered in the NPM. The percentages of missing data on material origin and age were higher in the congenital metabolic disorders registers, although the proportion of births by maternal origin and the mean maternal age in each ethnic group was fairly similar to that in the NPM. The percentages of missing data on birthweight and gestational age were much higher in the NPM data than in the congenital metabolic disorders registers, especially among births registered in Catalonia and births to foreign mothers. CONCLUSIONS: Our results suggest some limitations in the quality of the data on gestational age and birthweight provided by NPM data, especially for comparisons of preterm and low birthweight indicators in the Spanish-born and immigrant populations. Moreover, the results point to the quality of the congenital metabolic disorders registers as a source to compare reproductive and perinatal health indicators. PMID- 20022146 TI - Neural computational prediction of oral drug absorption based on CODES 2D descriptors. AB - A neural model based on a numerical molecular representation using CODES program to predict oral absorption of any structure is described. This model predicts both high and low-absorbed compounds with a global accuracy level of 74%. CODES/ANN methodology shows promising utilities not only as a conventional in silico tool in high-throughput screening or improvement of absorption capabilities procedures but also the improvement of in vitro-in vivo correlation could be addressed. PMID- 20022147 TI - The shifting foundations of nursing. AB - In this paper we argue that the concerns generated by the development of Foundation Degrees and the Assistant and Associate Practitioner roles have rekindled some of the unresolved debates regarding the status and identity of nursing and nurses. Through the application of the sociological theories of professionalisation and nostalgia we have identified the shifting and unresolved nature of nursing. We argue that these theories continue to have resonance in the current climate of change and 'upskilling' of the health care workforce and argue, that the shifts illuminated are perhaps so significant as to demonstrate that we have entered a post-nursing era. PMID- 20022148 TI - Cross-border radon index map 1:100 000 Lausitz - Jizera - Karkonosze - Region (northern part of the Bohemian Massif). AB - The first cross-border map describing the radon (Rn) risk from bedrock was assembled in the northern part of the Bohemian Massif at a scale 1:100 000. The map covers the area of Lausitz (Germany), Karkonosze (Czech Republic and Poland) and Jizera (Czech Republic). The map is based on 818 measurements of soil gas Rn in rock types of Precambrian to Mesozoic age with variable geology. Geographic information system (GIS) processing enabled a good coincidence of soil gas Rn concentrations between data from all three countries in lithologically adjacent rock types as well as the direct correlation to georeferenced indoor Rn values, which was tested using the Czech indoor Rn data. The method of data processing can contribute to assembling the European Geogenic Radon Map. PMID- 20022149 TI - An approach to improve the Austrian Radon Potential Map by Bayesian statistics. AB - A Radon Potential Map as well as a mean indoor Radon Concentration Map is available from the Austrian National Radon Project (1992-2002). These maps are based on the average Radon Potential/Concentration within every municipality and they sort municipalities into three radon 'risk' classes. This is a convenient way for the administration, but it does not describe the real radon risk distribution within a municipality because of the often inhomogeneous geological situation. Therefore, a combination of indoor radon data with all relevant parameters such as house type, storey and ventilation rates along with geological information should be used to improve the existing radon maps. The method, described here, uses Bayes' theory to combine the Radon Potential derived from indoor radon measurements with information from geology. The existing Radon Potential Map was improved by using available soil gas radon data at certain geological units and extrapolated transfer factors. The modifications of the map are shown and several problems arising with the application of this technique are discussed. PMID- 20022150 TI - The role of alcohol in oral precancer: observations from a North-East England population. AB - Alcohol is known to be a risk factor for oral precancerous lesions, but evidence has been weakened by subjective estimates of alcohol intake from patients, and confounded by their use of tobacco. Red cell macrocytosis, assessed by calculation of mean corpuscular volume (MCV), may be a useful objective indicator of chronic alcohol intake. The aim of this study was to compare subjective and objective measures of alcohol intake in patients with oral precancerous lesions and assess the use of reported alcohol intake and MCV on assessing the degree of dysplasia at presentation and their role as markers of the behaviour of such lesions by assessing clinical outcome after treatment. Fifty-four new patients were recruited. All were smokers and had histologically confirmed single dysplastic oral precancerous lesions, but had had no previous treatment. Subjective data about their alcohol consumption were recorded, and blood samples taken for the assessment of MCV. All patients had laser excision of their lesions, which were assessed histopathologically for signs of dysplasia. Patients were followed up for 2 years. The significance of differences was assessed using Fisher's exact test. Alcohol intake of more than 28 units/week and MCV of over 100 were associated with increased dysplasia at presentation (p=0.01 and p=0.03, respectively). Thirty-six patients were disease-free at 2 years, but 18 developed further disease, often at new sites. Alcohol intake of more than 28 units/week was significantly associated with an increased risk of further disease (p=0.03), particularly recurrence at the same site (p=0.02). PMID- 20022151 TI - Facing the Future: evidence from Joint Aspen FACE, SoyFACE and SFB 607 meeting. PMID- 20022152 TI - The role of hydrostatic pressure on developmental stages of Pomatoceros lamarcki (Polychaeta: Serpulidae) exposed to water accommodated fractions of crude oil and positive genotoxins at simulated depths of 1000-3000 m. AB - The effect of high hydrostatic pressures on the ecotoxicological profile of pollutants is an unexplored research area. Using Pomatoceros lamarcki as a surrogate organism for this eco-barotoxicological study, it was found that in a 48 h larval bioassay with water accommodated fractions (WAF) of crude oil of up to 15.1 mg L(-1) (total hydrocarbon content) and hydrostatic pressures up to 300 bar (3000 m), an additive response was found (p < 0.001) rather than any synergism (p = 0.881). Comprehensive cytogenetic analysis of 6-h (15 degrees C) embryos exposed to WAF (0.19 mg L(-1)) at 100 bar showed no effects on mitotic fidelity or cell division rate over the 1 bar treatment. However, embryo's treated with the clastogen mitomycin-c at 100 bar exhibited a significant increase in mitotic aberrations over 1 bar treated as was the case with hypo/hypersaline treatments (p < 0.05). Conversely, an increase in hydrostatic pressure actually reduced the effects of spindle inhibition by the aneugen colchicine (p < 0.05). PMID- 20022153 TI - Tree-ring stable isotopes and historical perspectives on pollution--an overview. AB - Hydrogen (delta2H), carbon (delta13C), oxygen (delta18O) and nitrogen (delta15N) isotopes of tree rings growing in field conditions can be indicative of past pollution effects. The characteristic delta13C trend is a positive shift generally explained by invoking closure of stomata, but experimental studies suggest that increased rates of carboxylation could also generate such trends. In many cases the delta18O and delta2H values decrease in trees exposed to pollution and exhibit inverse coinciding long-term trends with delta13C values. However, some trees exposed to diffuse pollution and experimental conditions can show an increase or no delta18O change even if delta13C values increase. These diverse responses depend on how stress conditions modify physiological functions such as stomatal conductance, carboxylation, respiration, and perhaps water assimilation by the root system. Recent studies suggest that delta15N changes in trees can be caused by soil acidification and accumulation of anthropogenic emissions with isotopic signals deviating from natural N. PMID- 20022154 TI - Uranium uptake and depuration in the aquatic invertebrate Chironomus tentans. AB - Evaluation of aqueous uranium (U) uptake and depuration in larvae of the midge Chironomus tentans were investigated in two separated experiments. First, a static-renewal experiment was performed with 10-d old C. tentans larvae exposed to 300 microg U/L. The animals steadily accumulated U (K(u) = 20.3) approaching steady-state conditions (BAF = 56) in approximately 9-11 d. However, accumulated U was readily depurated (K(d) = 0.36) with U tissue concentration decreasing rapidly within 3 d of the larvae being placed in clean water (t(1/2) = 1.9 d). Also, the growth of C. tentans larvae appeared to decrease after 6-11 d of U exposure, probably due to the reallocation of resources into U detoxification mechanisms. However, growth significantly increased once C. tentans were transferred to clean water. A separate short-term experiment was performed to evaluate the possible mechanism of U uptake in this invertebrate. Results suggested a passive mechanism of U uptake coupled with an active mechanism of U depuration but no details related to the type of mechanisms or pathway was investigated. PMID- 20022155 TI - The rhizosphere and PAH amendment mediate impacts on functional and structural bacterial diversity in sandy peat soil. AB - To reveal the degradation capacity of bacteria in PAH polluted soil and rhizosphere we combined bacterial extradiol ring-cleavage dioxygenase and 16S rRNA analysis in Betula pubescens rhizoremediation. Characterisation of the functional bacterial community by RFLP revealed novel environmental dioxygenases, and their putative hosts were studied by 16S rRNA amplification. Plant rhizosphere and PAH amendment effects were detected by the RFLP/T-RFLP analysis. Functional species richness increased in the birch rhizosphere and PAH amendment impacted the compositional diversity of the dioxygenases and the structural 16S rRNA community. A shift from an Acidobacteria and Verrucomicrobia dominated to an Alpha- and Betaproteobacteria dominated community structure was detected in polluted soil. Clone sequence analysis indicated catabolic significance of Burkholderia in PAH polluted soil. These results advance our understanding of rhizoremediation and unveil the extent of uncharacterized functional bacteria to benefit bioremediation by facilitating the development of the molecular tool box to monitor bacterial populations in biodegradation. PMID- 20022156 TI - Anisocoria after open reduction and internal fixation of a mandible fracture under general anesthesia: a case report. PMID- 20022157 TI - Time to invasive airway placement and resuscitation outcomes after inhospital cardiopulmonary arrest. AB - BACKGROUND: Clinicians often place high priority on invasive airway placement during cardiopulmonary resuscitation. The benefit of early vs. later invasive airway placement remains unknown. In this study we examined the association between time to invasive airway (TTIA) placement and patient outcomes after inhospital cardiopulmonary arrest (CPA). METHODS: We analyzed data from the National Registry of Cardiopulmonary Resuscitation (NRCPR). We included hospitalized adult patients receiving attempted invasive airway placement (endotracheal intubation, laryngeal mask airway, tracheostomy, and cricothyrotomy) after the onset of CPA. We excluded cases in which airway insertion was attempted after return of spontaneous circulation (ROSC). We defined TTIA as the elapsed time from CPA recognition to accomplishment of an invasive airway. The primary outcomes were ROSC, 24-h survival, and survival to hospital discharge. We used multivariable logistic regression to evaluate the association between the patient outcome and early (<5 min) vs. later (> or =5 min) TTIA, adjusted for hospital location, patient age and gender, first documented pulseless ECG rhythm, precipitating etiology and witnessed arrest. RESULTS: Of 82,649 CPA events, we studied the 25,006 cases in which TTIA was recorded and the inclusion criteria were met. Observations were most commonly excluded for not having an invasive airway emergently placed during resuscitation. The mean time to invasive airway placement was 5.9 min (95% CI: 5.8-6.0). Patient outcomes were: ROSC 50.3% (49.7-51.0%), 24-h survival 33.7% (33.1-34.3%), survival to discharge 15.3% (14.9-15.8%). Early TTIA was not associated with ROSC (adjusted OR: 0.96, 0.91-1.01) but was associated with better odds of 24-h survival (adjusted OR: 0.94, 0.89-0.99). The relationships between TTIA and survival to discharge could not be determined. CONCLUSIONS: Early invasive airway insertion was not associated with ROSC but was associated with slightly improved 24-h survival. Early invasive airway management may or may not improve inhospital cardiopulmonary resuscitation outcomes. PMID- 20022158 TI - Wheezing and pneumonia: a complex relationship. PMID- 20022159 TI - Increased plasma metastin levels in adolescent women with polycystic ovary syndrome. AB - OBJECTIVE: This study was designed to: (1) measure metastin levels in women with polycystic ovary syndrome (PCOS) and in adolescent controls; (2) investigate the possible correlations between metastin and PCOS-related reproductive and metabolic disturbances. STUDY DESIGN: The study was a clinical study. Nineteen adolescent women with PCOS, twenty-three adult women with the syndrome, and twenty adolescent controls were selected. Blood samples were collected between day 1 and day 5 of a spontaneous bleeding episode in the PCOS groups and of a menstrual cycle of the controls at 9 a.m. after an overnight fast. Circulating levels of LH, FSH, prolactin, testosterone (T), free testosterone, DHEAS, sex hormone-binding globulin, insulin, glucose and metastin were measured. RESULT(S): Plasma metastin levels are increased in adolescent women with PCOS compared to adolescent controls. Plasma metastin levels were positively correlated with LH levels, 2-h glucose levels and T levels. CONCLUSION(S): These results indicate that metastin is increased in adolescent PCOS women. The increased metastin levels were positively correlated with LH and T levels, and may affect the development of PCOS in adolescents. PMID- 20022160 TI - Effects of landscape protection on farm management and farmers' income in Norway. AB - Protected Landscapes (PLs) are increasingly used in Norway to conserve cultural (human modified) landscapes. In many cases the maintenance of agricultural activities in PLs is required to preserve landscape character. Whilst research exists on land conservation policies in general, the particular effects of PL on management and adjustment of the farms involved have not received attention in the literature. We present results from a questionnaire sent to owners of agricultural land within PLs in Norway. Whilst landowners were divided upon the effects of PLs on farm management, the economic situation of the farm was little affected. Furthermore, changes in farm management after the establishment of a PL did not seem to have been driven by the establishment of the PLs per se. Most importantly, farm management changes were related to potential options to develop the farm and its land. A statistical model showed that PL-farms did not differ significantly from farms outside PL in the development of their land use or animal husbandry. Our findings thus indicate that the establishment of PL played a minor role as a driving force of changes in farm management and farm income. PMID- 20022161 TI - A multi-criteria assessment of fishing gear impacts in demersal fisheries. AB - Fishing gears have multiple impacts on the marine environment, and policies to reduce these impacts through modifying fishing gears are becoming common place. Different modifications result in different changes in the set of environmental impacts, and imply different sets of costs and benefits for different stakeholder groups. In this study, the analytic hierarchy process (AHP) is used to quantify the relative importance of the environmental impacts of fishing to different stakeholder groups. Forty eight individuals representing six different stakeholder groups (ecologists, biologists, economists, gear technologists, fishers and fisheries managers) were surveyed. As expected, fishers and gear technologists placed substantially greater importance on reducing discarding of commercial fish species than on habitat damage. Priorities of other stakeholder groups varied, but all placed greater priority on habitats than the commercial sector. The results suggest that management aimed at reducing environmental impacts of fishing broader than just discarding is appropriate, but such moves are likely to be opposed by the fishing industry. The derived weights also have a direct application to fisheries management, as they allow otherwise non commensurate impacts to be aggregated into an overall impact to compare environmental benefits from alternative modifications of fishing gear. PMID- 20022162 TI - Differential complication rates following radical cystectomy in the irradiated and nonirradiated pelvis. AB - BACKGROUND: Reports suggest that cystectomy following pelvic irradiation is associated with a higher morbidity and mortality than in primary cases. However, such reports are from an era when postcystectomy complication rates were higher than are currently reported. OBJECTIVE: This study evaluates perioperative complications and mortality in primary radical and postradiation salvage cystectomy. DESIGN, SETTING, AND PARTICIPANTS: Patients treated with cystectomy for bladder cancer or advanced pelvic malignancies involving the bladder were studied. MEASUREMENTS: Perioperative complications and mortality were analysed for 426 primary and 420 salvage cystectomies performed at a single institution between 1970 and 2005. RESULTS AND LIMITATIONS: The 30- and 60-d mortality in the 2000-2005 cohort were 0% and 1.2%, respectively, in the primary group and 1.4% and 4.3%, respectively, in the salvage cystectomy group. Thirty-day mortality between 1970 and 2005 was not statistically significant in the primary and salvage groups (4.2% and 7.1%, respectively). CONCLUSIONS: This large series from a high-volume centre demonstrates no difference in perioperative mortality in primary or postradiation salvage radical cystectomy. Similarly, there was no significant difference in the incidence of most of the surgical or medical complications in either group, although the stomal stenosis rate was higher postradiation. PMID- 20022163 TI - Health risk assessment of BTEX emissions in the landfill environment. AB - This study focuses on a health risk assessment related to benzene, toluene, ethylbenzene, and m,p,o-xylene (xylenes) (BTEX) exposure via inhalation for workers at a landfill (LF) site. First, the landfill gas (LFG) samples were collected and analyzed accordance with US EPA method TO-17. The mean concentrations of benzene, toluene, ethylbenzene, and xylenes were determined as 140.3, 1271.7, 239.9, and 341.3 microg/m(3), respectively. Then, a risk assessment methodology was employed to evaluate the potential adverse health effects of the individual BTEX compounds according to their carcinogenicities. The corresponding mean cancer risk for benzene was estimated to be 6.75E-05 that is lower than the designated acceptable risk level of 1.0E-04. With respect to mean non-carcinogenic risks for toluene, ethylbenzene, and xylenes, both individually and cumulatively, they were lower than the specified level of 1.0. These findings reveal that landfill BTEX emissions do not pose a health threat to workers at the landfill site. In addition, as far as the risks are concerned for the population in the neighborhood area of the landfill, air dilution of BTEX emitted from LF site is widely sufficient to guarantee their protection. PMID- 20022165 TI - Enhanced phenol-photodegradation by particulate semiconductor mixtures: interparticle electron-jump. AB - Degradation of phenol on suspended TiO(2), ZnO, CdO, Fe(2)O(3), CuO, ZnS and Nb(2)O(5) particles under UV-A light exhibit identical photokinetic behavior; follow first-order kinetics, display linear dependence on the photon flux and slowdown with increase of pH. All the semiconductors show sustainable photocatalytic activity. Dissolved O(2) is essential for the photodegradation and oxidizing agents like H(2)O(2), Na(2)BO(3), K(2)S(2)O(8), KBrO(3), KIO(3) and KIO(4), reducing agents such as NaNO(2) and Na(2)SO(3) and sacrificial electron donors like hydroquinone, diphenyl amine and trimethyl amine enhance the degradation. However, the photocatalysis is insensitive to pre-sonication. Two particulate semiconductors present together, under suspension and at continuous motion, enhance the photocatalytic degradation up to about four-fold revealing interparticle electron-jump. PMID- 20022164 TI - Total oxidation of toluene on Pt/CeO2-ZrO2-Bi2O3/gamma-Al2O3 catalysts prepared in the presence of polyvinyl pyrrolidone. AB - Pt/CeO(2)-ZrO(2)-Bi(2)O(3)/gamma-Al(2)O(3) (Pt/CZB/Al(2)O(3)) catalysts for the catalytic combustion of toluene, which is one of the volatile organic compounds (VOCs), were prepared by the wet impregnation method in the presence of polyvinyl pyrrolidone (PVP). X-ray powder diffraction, transmission electron microscopy, and BET specific surface area measurement using N(2) adsorption have been used to characterize the catalysts. The catalytic test was conducted from room temperature in a flow of 900 ppm of toluene in air and gas hourly space velocity (GHSV) of 8000 h(-1). The catalytic activity was evaluated in terms of C(7)H(8) conversion and the gas composition after the reaction was analyzed using two gas chromatographs with a flame ionization detector (FID) and a thermal conductivity detector (TCD). The Pt/CZB/Al(2)O(3) catalysts are specific for the total toluene oxidation and CO and any toluene-derivative compounds were not detected as by products. The specific surface area of the catalysts was increased by the addition of PVP in the preparation process. By the optimization of the amount of platinum, complete oxidation of toluene was realized at a temperature as low as 120 degrees C on a 7 wt%Pt/16 wt%Ce(0.64)Zr(0.15)Bi(0.21)O(1.895)/gamma-Al(2)O(3) catalyst. PMID- 20022166 TI - Statistical optimization of process parameters for landfill leachate treatment using electro-Fenton technique. AB - Mature landfill leachate is typically non-biodegradable and contains high concentration of refractory organics. The aim of this research was to optimize operating parameters in electro-Fenton process, for the removal of recalcitrant organics from semi-aerobic landfill leachate using response surface methodology (RSM). Effectiveness of important process parameters H(2)O(2)/Fe(2+) molar ratio, current density, pH and reaction time were determined, optimized and modeled successfully. Significant quadratic polynomial models were obtained (R(2)=0.9972 and 0.9984 for COD and color removals, respectively). Numerical optimization based on desirability function were employed; in a 43 min trial 94.07% of COD and 95.83% of color were removed at pH 3 and H(2)O(2)/Fe(2+) molar ratio 1, while current density was 49 mA/cm(2). The results indicate that E-Fenton process was an effective technology for semi-aerobic landfill leachate treatment. PMID- 20022167 TI - A novel estrogenic compound transformed from fenthion under UV-A irradiation. AB - The photo-transformed products of fenthion well-known as one of the most photosensitive organophosphorus insecticides and their estrogenic activities were investigated using a yeast two-hybrid assay incorporating the human estrogen receptor alpha (hERalpha). We identified fenthion sulfoxide and 3-methyl-4 methylsulfinylphenol (MMS) as the major transformed products and 3-methyl-4 (methylthio)phenol (MMP) as the minor product under UV-A irradiation. Further, significant estrogenic activity was observed in the solution irradiated for 160 min; this activity was evaluated as 18 pM converted to 17beta-estradiol (E(2)) equivalent concentration. By using authentic standards, it was found that MMP possessed weak estrogenic activity; its activity was evaluated as 1.7x10(-6) times compared with that of E(2). However, it was also revealed that the activity due to MMP was only 13%. From high-performance liquid chromatography (HPLC) and nuclear magnetic resonance (NMR) spectroscopies, we newly identified a significant estrogenic compound transformed from fenthion, O,O-dimethyl S-[3 methyl-4-(methylthio)phenyl]phosphorothioate, S-aryl fenthion. PMID- 20022168 TI - Using orthogonal design to determine optimal conditions for biodegradation of phenanthrene in mangrove sediment slurry. AB - In the present paper, the effects of four factors, each at three levels, on biodegradation of phenanthrene, a 3-ring PAH, in contaminated mangrove sediment slurry were investigated using the orthogonal experimental design. The factors and levels were (i) sediment types (clay loam, clayey and sandy); (ii) different inoculums (Sphingomonas sp., a mixture of Sphingomonas sp. and Mycobacterium sp., and without inoculum); (iii) presence of other PAHs (fluorene, pyrene, and none); and (iv) different salinities (5, 15 and 25 ppt). Variance analysis based on the percentages of Phe biodegradation showed that the presence of other PAHs had little effect on phenanthrene biodegradation. The kinetics of phenanthrene biodegradation in all experiments was best fitted by the first order rate model. The highest first order rate constant, k value was 0.1172 h(-1) with 97% Phe degradation; while the lowest k value was 0.0004 and phenanthrene was not degraded throughout the 7-d experiment. The p values of k for the four factors followed the same trend as that for the biodegradation percentage. Difference analysis revealed that optimal phenanthrene biodegradation would take place in clay loam sediment slurry at low salinity (5 to 15 ppt) with the inoculation of both Sphingomonas sp. and Mycobacterium sp. PMID- 20022169 TI - Removal of 2,4-diclorophenol from aqueous solutions using tobacco hairy root cultures. AB - 2,4-Dichlorophenol (2,4-DCP) is harmful for aquatic life and human health, so many attempts have focused on removing it through innocuous technologies. Hairy roots (HR) represent an interesting plant system to study the process and to remove efficiently this compound. In the present work, tobacco HR clones were obtained and one of them was selected for 2,4-DCP phytoremediation assays. These cultures removed 2,4-DCP in short time and with high efficiency (98%, 88% and 83%) for solutions initially containing 250, 500 and 1000 mg/L, respectively. Removal process was mainly associated with peroxidase activity. The highest efficiency for 2,4-DCP (500 mg/L) removal was reached at 60 min and using 10 mM H(2)O(2). Moreover, HR could be re-used, almost for three consecutive cycles. The diminution of pH and the increase of chloride ions in post-removal solutions suggested that 2,4-DCP dehalogenation was mediated by peroxidases. Moreover, changes in deposition pattern of lignin in HR exposed to 2,4-DCP suggested that cell walls of xylem and phloem elements would be the site of deposition of some products formed and they would be a lignin-type polymer. These findings contribute to understand 2,4-DCP removal process with tobacco HR and it might have implications in the use of this system for decontamination of polluted waters. PMID- 20022170 TI - Production of geopolymers using glass produced from DC plasma treatment of air pollution control (APC) residues. AB - Air pollution control (APC) residues are the hazardous waste produced from cleaning gaseous emissions at energy-from-waste (EfW) facilities processing municipal solid waste (MSW). APC residues have been blended with glass-forming additives and treated using DC plasma technology to produce a high calcium alumino-silicate glass. This research has investigated the optimisation and properties of geopolymers prepared from this glass. Work has shown that high strength geopolymers can be formed and that the NaOH concentration of the activating solution significantly affects the properties. The broad particle size distribution of the APC residue glass used in these experiments results in a microstructure that contains unreacted glass particles included within a geopolymer binder phase. The high calcium content of APC residues may cause the formation of some amorphous calcium silicate hydrate (C-S-H) gel. A mix prepared with S/L=3.4, Si/Al=2.6 and [NaOH]=6M in the activating solution, produced high strength geopolymers with compressive strengths of approximately 130 MPa. This material had high density (2070 kg/m(3)) and low porosity. The research demonstrates for the first time that glass derived from DC plasma treatment of APC residues can be used to form high strength geopolymer-glass composites that have potential for use in a range of applications. PMID- 20022171 TI - Effective nitrosamines trap derived from the in situ carbonized mesoporous silica MCM-41. AB - A new route to modify the mesoporous silica MCM-41 with carbon, using the inherent surfactant template in the as-synthesized sample as the carbon precursor, is reported in this article. Apart from the advantage of omitting energy and time required for removal of template, the resulting silica-carbon composites exhibit a high efficiency in adsorption and catalytic decomposition of N-nitrosopyrrolidine (NPYR), the typical carcinogenic pollutant in environment. The influence of carbonization temperature on the structure and performance of the resulting carbon-silica composite was investigated by use of XRD, N(2) adsorption-desorption, FTIR and TG-DSC techniques, and the instantaneous adsorption and the temperature-programmed surface reaction (TPSR) of NPYR as well as the reduction of nitrosamines level of cigarette smoke were also utilized to assess the actual function of these composites. The carbonization performed in the range of 773-973 K enabled the resulting composites to possess a higher activity than parent MCM-41 in eliminating volatile nitrosamines, which is beneficial for controlling carcinogenic pollutants in environment. PMID- 20022172 TI - Coprecipitation of trace elements with Ni2+/2-Nitroso-1-naphthol-4-sulfonic acid and their determination by flame atomic absorption spectrometry. AB - Ni(2+)/2-Nitroso-1-naphthol-4-sulfonic acid precipitate was used for the coprecipitation of Co, Pb, Cu, Fe and Zn prior to their flame atomic absorption spectrometric (FAAS) determinations in environmental samples. The precipitate could be easily dissolved with concentrated nitric acid. The recovery values for analyte ions were higher than 95%. The parameters including pH, sample volume, centrifuge time, amounts of nickel and matrix effects were optimized for the quantitative recoveries of the analytes. The relative standard deviations of cobalt, lead, copper, iron and zinc were found 4.5, 5.7, 3.8, 6.1 and 7.5%, respectively. The limit of detection was calculated as 1.05, 2.67, 1.30, 1.38, and 0.50 microg L(-1) for cobalt, lead, copper, iron and zinc. The validation of the procedure was checked by the analysis of IAEA 336 lichen and SLRS 4 Riverine water standard reference materials were analyzed with satisfactory results. The presented coprecipitation procedure was successfully applied to some environmental samples for determination of analyte ions. PMID- 20022173 TI - Heavy metal sources and associated risk in response to agricultural intensification in the estuarine sediments of Chaohu Lake Valley, East China. AB - In this study, the source and risk associated with heavy metals in the estuarine sediments of Chaohu Lake Valley in East China were studied. The results demonstrated that most metal contents (Cd, Pb and Zn) increased continuously from 0.21, 8.05 and 73.76 mg/kg in the substrate to 0.33, 17.20 and 100.22 mg/kg in the surface sediments on average, respectively, in all ten estuaries evaluated. In addition, the high proportion of chemically reactive forms of Cd (18.10 33.60%) implied a moderately high risk, while the risk associated with Pb and Zn was moderate in most estuaries. Enrichment factor (EF) and cluster analysis (CA) were conducted to assess the metal sources. The EF values of Cd, Pb and Zn (4.96, 3.68 and 2.25, respectively) indicated that they had accumulated in the estuarine sediments. Additionally, CA suggested that their primary sources were anthropogenic, namely the extensive use of fertilizers. Therefore, heavy metal pollution due to agricultural intensification in Chaohu Lake Valley should be taken into account during the formation of valley management strategies for the water environment. PMID- 20022174 TI - Assessment of the BCR sequential extraction procedure for thallium fractionation using synthetic mineral mixtures. AB - This work focused on the specific behavior of Tl-bearing phases in the BCR (Community Bureau of Reference) sequential extraction (SE) scheme, namely Tl bearing ferrihydrite, goethite, birnessite, calcite, illite, sphalerite and feldspar in their simple model mixtures with quartz. Several significant discrepancies between the obtained and expected behaviors of these phases in the BCR SE were observed. The amount of Tl released as the exchangeable/acid extractable fraction (55-82% of the total Tl content) showed a substantial H(+) promoted dissolution of all Fe(III) and Mn(III, IV) oxides (corresponding to up to 61% of solid Fe dissolved) and incongruent (increased) extraction of Tl from ferrihydrite and goethite. Reductive conditions of the second SE step were insufficient to complete goethite dissolution with corresponding Tl amount retained in the solid phase. Similarly, insufficient oxidation of sphalerite and lower Tl recovery of the oxidisable fraction was identified. In contrast, the BCR SE seems to produce well predictable results of Tl leaching from Tl-bearing calcite and feldspar. Only 70% of total Tl content was extracted from Tl-modified illite in the exchangeable/acid-extractable step, while 30% was associated with the reducible and residual fractions, i.e., Tl was strongly fixed to the illite matrix. PMID- 20022175 TI - Treatment efficiency and VFA composition of a thermophilic anaerobic contact reactor treating food industry wastewater. AB - The objective of this study was to examine the effects of organic loading rate and hydraulic retention time on volatile fatty acid composition and treatment efficiency of high rate thermophilic anaerobic contact reactor (TACR) treating potato-chips wastewaters. The operational performance of TACR was monitored from start-up by assessing chemical oxygen demand (COD) removal efficiencies, volatile fatty acid (VFA) production and biogas composition. The reactor was studied at different organic loading rates (OLRs) ranging from 0.6 to 8 kg COD/m(3) d. The COD removal efficiencies were found to be 86-97% and the methane percentage of the biogas produced was 68-89% during the OLRs studied. The approximate methane yield was found to be 0.42 m(3) CH(4)/kg COD(removed). The major intermediate products of anaerobic digestion were acetate, propionate, iso-butyrate, butyrate, iso-valerate, valerate, iso-caproate and caproate. The use of thermophilic anaerobic contact reactor offers a sustainable technology for the treatment of potato-chips wastewaters since high COD removal efficiencies and high methane percentage in the biogas produced can be attained even at high OLRs. PMID- 20022177 TI - Simulation of scanning transmission electron microscope images on desktop computers. AB - Two independent strategies are presented for reducing the computation time of multislice simulations of scanning transmission electron microscope (STEM) images: (1) optimal probe sampling, and (2) the use of desktop graphics processing units. The first strategy is applicable to STEM images generated by elastic and/or inelastic scattering, and requires minimal effort for its implementation. Used together, these two strategies can reduce typical computation times from days to hours, allowing practical simulation of STEM images of general atomic structures on a desktop computer. PMID- 20022176 TI - The c-Jun N-terminal kinase 1 (JNK1) in spinal astrocytes is required for the maintenance of bilateral mechanical allodynia under a persistent inflammatory pain condition. AB - Peripheral inflammation induces persistent central sensitization characterized by mechanical allodynia and heat hyperalgesia that are mediated by distinct mechanisms. Compared to well-demonstrated mechanisms of heat hyperalgesia, mechanisms underlying the development of mechanical allodynia and contralateral pain are incompletely known. In this study, we investigated the distinct role of spinal JNK in heat hyperalgesia, mechanical allodynia, and contralateral pain in an inflammatory pain model. Intraplantar injection of complete Freund's adjuvant (CFA) induced bilateral mechanical allodynia but unilateral heat hyperalgesia. CFA also induced a bilateral activation (phosphorylation) of JNK in the spinal cord, and the phospho JNK1 (pJNK1) levels were much higher than that of pJNK2. Notably, both pJNK and JNK1 were expressed in GFAP-positive astrocytes. Intrathecal infusion of a selective peptide inhibitor of JNK, D-JNKI-1, starting before inflammation via an osmotic pump, reduced CFA-induced mechanical allodynia in the maintenance phase but had no effect on CFA-induced heat hyperalgesia. A bolus intrathecal injection of D-JNKI-1 or SP600126, a small molecule inhibitor of JNK also reversed mechanical allodynia bilaterally. In contrast, peripheral (intraplantar) administration of D-JNKI-1 reduced the induction of CFA-induced heat hyperalgesia but did not change mechanical allodynia. Finally, CFA-induced bilateral mechanical allodynia was attenuated in mice lacking JNK1 but not JNK2. Taken together, our data suggest that spinal JNK, in particular JNK1 plays an important role in the maintenance of persistent inflammatory pain. Our findings also reveal a unique role of JNK1 and astrocyte network in regulating tactile allodynia and contralateral pain. PMID- 20022178 TI - Noise in off-axis type holograms including reconstruction and CCD camera parameters. AB - Phase and amplitude images as contained in digital holograms are commonly extracted via a process called "reconstruction". Expressions for the expected noise in these images have been given in the past by several authors; however, the effect of the actual reconstruction process has not been fully appreciated. By starting with the Quantum Mechanical intensity distribution of the off-axis type interference pattern, then building the digital hologram on an electron-by electron base while simultaneously reconstructing the phase/amplitude images and evaluating their noise levels, an expression is derived that consistently describes the noise in simulated and experimental phase/amplitude images and contains the reconstruction parameters. Because of the necessity to discretize the intensity distribution function, the digitization effects of an ideal CCD camera had to be included. Subsequently, this allowed a comparison between real and simulated holograms which then led to a comparison between the performance of an "ideal" CCD camera versus a real device. It was concluded that significant improvement of the phase and amplitude noise may be obtained if CCD cameras were optimized for digitizing intensity distributions at low sampling rates. PMID- 20022179 TI - Cs corrected STEM EELS: Analysing beam sensitive carbon nanomaterials in cellular structures. AB - Identification of individual single wall nanotubes (SWNTs) within a cellular structure can provide vital information towards understanding the potential mechanisms of uptake, their localisation and whether their structure is transformed within a cell. To be able to image an individual SWNT in such an environment a resolution is required that is not usually appropriate for biological sections. Standard transmission electron microscopy (TEM) techniques such as bright field imaging of these cellular structures result in very weak contrast. Traditionally, researchers have stained the cells with heavy metal stains to enhance the cellular structure, however this can lead to confusion when analysing the samples at high resolution. Subsequently, alternative methods have been investigated to allow high resolution imaging and spectroscopy to identify SWNTs within the cell; here we will concentrate on the sample preparation and experimental methods used to achieve such resolution. PMID- 20022180 TI - Near-field scanning optical microscopy using polymethylmethacrylate optical fiber probes. AB - We report the first use of polymethylmethacrylate (PMMA) optical fiber-made probes for scanning near-field optical microscopy (SNOM). The sharp tips were prepared by chemical etching of the fibers in ethyl acetate, and the probes were prepared by proper gluing of sharpened fibers onto the tuning fork in the conditions of the double resonance (working frequency of a tuning fork coincides with the resonance frequency of dithering of the free-standing part of the fiber) reported earlier for the case of glass fibers. Quality factors of the probes in the range 2000-6000 were obtained, which enables the realization of an excellent topographical resolution including state-of-art imaging of single DNA molecules. Near-field optical performance of the microscope is illustrated by the Photon Scanning Tunneling Microscope images of fluorescent beads with a diameter of 100nm. The preparation of these plastic fiber probes proved to be easy, needs no hazardous material and/or procedures, and typical lifetime of a probe essentially exceeds that characteristic for the glass fiber probe. PMID- 20022182 TI - Autonomic denervation and the origins of chronic Western diseases. AB - Many chronic Western diseases result from lifestyles that include refined diets, poor bowel habits, limited physical exercise and suboptimal patterns of childbirth. Western diets result in reduced stool weights, increased bowel transit times and persistent physical efforts during defaecation. Prolonged physical efforts during defaecation and childbirth cause latent, or direct, injuries to branches of the cardiac (thorax), coeliac (abdomen) and hypogastric (pelvis) plexi. Injuries to autonomic nerves result in impaired visceral function including visceral dysmotility, tissue hypoplasia and hyperplasia, increased susceptibility to infection, and, aberrant reinnervation with sensitisation of the central nervous system (CNS). These unrecognised injuries are vulnerable to the long list of causes of autonomic Dysfunction, e.g. stress, alcohol, drugs, infection, trauma, cancer, etc. Specific injuries at different anatomical locations in midline autonomic pathways give rise to a wide range of Western diseases from infancy to old age, through diverse and cumulative mechanisms. PMID- 20022181 TI - Cortisol function among early school-aged homeless children. AB - Homelessness represents a context of extreme poverty and risk for child development. This study compared the relative influence of two classes of risk in the context of homelessness. Levels of socioeconomic resource-related risk and negative lifetime events were examined with respect to morning cortisol levels and cortisol response to a set of cognitive tasks. Participants were 66 children between the ages of 4 and 7 years staying in an emergency shelter for families. Adversities largely reflecting family level negative life events predicted higher levels of morning cortisol and differences in initial level and change over the course of the session of cognitive tasks. In contrast, a socioeconomic cumulative risk score was not associated with morning or session-related differences in cortisol. PMID- 20022183 TI - Empirically defined subtypes of alcohol dependence in an Irish family sample. AB - Alcohol dependence (AD) is clinically and etiologically heterogeneous. The goal of this study was to explore AD subtypes among a sample of 1221 participants in the Irish Affected Sib Pair Study of Alcohol Dependence, all of whom met DSM-IV criteria for AD. Variables used to identify the subtypes included major depressive disorder, antisocial personality disorder, illicit drug dependence (cannabis, sedatives, stimulants, cocaine, opioids, and hallucinogens), nicotine dependence, the personality traits of neuroticism and novelty seeking, and early alcohol use. Using latent class analysis, a 3-class solution was identified as the most parsimonious description of the data. Individuals in a Mild class were least likely to have comorbid psychopathology, whereas a severe class had highest probabilities of all comorbid psychopathology. The third class was characterized by high probabilities of major depression and higher neuroticism scores, but lower likelihood of other comorbid disorders than seen in the severe class. Overall, sibling pair resemblance for class was stronger within than between classes, and was greatest for siblings within the severe class, suggesting a stronger familial etiology for this class. These findings are consistent with the affective regulation and behavioral disinhibition subtypes of alcoholism, and are in line with prior work suggesting familial influences on subtype etiology. PMID- 20022184 TI - Mortality among drug users after discharge from inpatient treatment: an 8-year prospective study. AB - BACKGROUND: Drug users who are leaving/completing inpatient medication-free treatment may, like drug users released from prison, have an elevated risk of dying from fatal overdoses. This is mainly explained by their low drug tolerance. METHODS: Two hundred and seventy-six drug users who had been admitted to 11 inpatient facilities in Norway, were followed prospectively after discharge from treatment during an 8-year period (1998-2006). The following instruments were used: EuropASI, SCL-25 and MCMI II. Information on deaths and causes of death were obtained from the National Death Register. RESULTS: A total of 36 deaths were registered after discharge from treatment during the observation period, of which 24 were classified as overdose deaths. During the first 4 weeks after discharge six persons died, yielding an unadjusted excess mortality of 15.7 (rate ratio) in this period (CI 5.3-38.3). All were dropouts and all deaths were classified as opiate overdoses. There was no significant association between time in index treatment and mortality after discharge, nor did any background characteristics correlate significantly with elevated mortality shortly after discharge. CONCLUSIONS: The elevated risk of dying from overdose within the first 4 weeks of leaving medication-free inpatient treatment is so dramatic that preventive measures should be taken. More studies from similar inpatient programmes are needed in order to obtain systematic knowledge about determinants of overdose deaths shortly after leaving treatment, and possible preventive measures. PMID- 20022185 TI - Virulence profile comparison between LEE-negative Shiga toxin-producing Escherichia coli (STEC) strains isolated from cattle and humans. AB - For comparison purposes, the prevalence of 8 virulence markers was investigated, by PCR, in 153 cattle and 47 human Locus for Enterocyte Effacement (LEE)-negative Shiga toxin-producing Escherichia coli (STEC) strains isolated in Argentina. Also, their correlation with severe disease was established. The virulence markers studied comprises 5 fimbrial and nonfimbrial adhesin-encoding genes (fimA, iha, efa1, lpfA(O113), and saa) and 3 toxin genes (cdt-V, subAB and astA) in addition to the Shiga toxins. The most prevalent virulence marker found was that encoded by the lpfA(O113) gene (199/200, 99%). Comparatively, the lpfA(O113), fimA, iha, saa, subAB, cdt-V and astA genes were detected in 100%, 92.8%, 85%, 52.9%, 36%, 11.8% and 9.8% of the cattle strains and in 97.9%, 95.7%, 89.4%, 40.4%, 32%, 17% and 10.6% of the human strains, respectively. All STEC strains were efa1 negative. The most prevalent profile observed among cattle and human STEC strains was lpfA(O113)iha fimA. These results show that bovine LEE negative STEC strains possessed genes encoding virulence factors present in human LEE-negative STEC strains that are associated with disease. Despite a great diversity of virulence profiles observed, further studies comparing wild type strains and their allelic mutants are needed to evaluate the role of each factor in the pathogenesis of LEE-negative STEC strains during human infections. PMID- 20022186 TI - Persistence of a Staphylococcus aureus small colony variants (S. aureus SCV) within bovine mammary epithelial cells. AB - Persistent bovine Staphylococcus aureus mastitis is attributable to the versatility of this pathogen within the mammary gland environment and to the formation of small colony variants (SCVs) that can survive within host cells. Previous studies had shown that S. aureus SCV Heba3231, isolated from a cow with chronic mastitis, had invaded and persisted in primary bovine aortic endothelial cells but caused minimal deleterious effects. The objective of this study was to investigate the interaction of SCV Heba3231 with bovine mammary epithelial cells (MAC-T cells) compared to its parent strain 3231 and to prototype strain Newbould 305. Monolayer cells were infected with each strain at various multiplicity of infections (MOIs) for 1 and 3.5h, followed by 20 min incubation with lysostaphin. Recovery of the SCV was significantly higher (P<0.05) after 3.8h with MOI of 100 compared to recovery of strains 3231 and Newbould 305. Upon further incubation, viable SCV were detected up to 96 h while 3231 were not isolated at 24h or later. Transmission electron microscopy demonstrated SCV uptake by MAC-T cells following a series of events similar to those for strain 3231. At 24h, multiple SCV were seen within enclosed vacuoles, while the 3231 parent strain was released extracellularly and the monolayer cells were damaged. The ability of SCV Heba3231 to survive inside vacuoles could be related to up-regulation of protective mechanisms. These findings highlight the potential role of bovine mammary epithelial cells and S. aureus SCV in persistent bovine mastitis. PMID- 20022187 TI - Markers of the uterine innate immune response of the mare. AB - Reproductive efficiency in mares is low and persistent mating-induced endometritis (PMIE) is an important cause of subfertility. Mating-induced endometritis (MIE) an obligate precursor to PMIE, is a ubiquitous, transient inflammatory response to the presence of sperm, seminal components and pathogens. However, the specific inflammatory pathways that derive from MIE and that may also be precursors to PMIE are not clear. The ability to identify and measure robust, repeatable markers of inflammation integral to MIE may be key to understanding the progression to PMIE. The aim of the study was to (i) refine a protocol for inducing MIE and in doing so test a range of cellular and molecular parameters as valid markers of MIE to facilitate future studies of mares susceptible to PMIE (ii) concurrently identify those parameters with potential as inflammatory indicators during MIE to inform and enhance early treatment regimens in practice. Mating-induced endometritis was induced in pony mares using a stringent protocol; mares were treated intrauterine with frozen/thawed semen (n = 5; FTS) or frozen/thawed extender (n = 6: FTEx). The parameters tested were measured before treatment were compared to samples collected at strategic time points after treatment: uterine cytology using cytological (at 8, 16, 24, 48 and 72 h after treatment) or histological analysis (at 24 and 72 h); uterine bacteriology (at 24 and 72 h); secretion of prostaglandin F(2alpha) (PGF(2alpha); at 8, 16, 24, 48 and 72 h); peripheral concentrations of serum amyloid A (SAA; at 24h); endometrial mRNA gene expression, focussing upon IL8 and TLR4, as examples of genes pertinent to inflammation (at 24 h). Uterine neutrophil cell numbers in both treatment groups increased at 8 (P < 0.001), 16 (P < 0.01) and 24 (P < 0.01) h after insemination, indicative of MIE and distinguished between different treatments because neutrophil numbers were greater from FTS mares than FTEx mares 8h after challenge. Uterine neutrophil cell numbers, assessed by histology, increased (P < 0.001) 24 and 72 h after treatment. Prostaglandin F(2alpha) concentrations increased (P < 0.05) 16 h after treatments, while SAA concentrations and bacterial growth scores were not significantly different after treatment. Endometrium from pony mares expressed mRNA for IL8 and TLR4 but expression was not altered after insemination. The protocol induced MIE, as confirmed by uterine cytology and maybe used hereafter as a repeatable and robust method for studying immune mechanisms that underlie MIE and so may aid the understanding of progression to persistent inflammation. It can be concluded that of the range of parameters tested, neutrophil cell numbers by cytological analysis and PGF(2alpha) were regarded as the most accurate markers of inflammation during MIE and important for use in practice. PMID- 20022189 TI - Revealing obliterated engraved marks on high strength aluminium alloy (AA7010) surfaces by etching technique. AB - Restoration of obliterated engraved marks on high strength Al-Zn-Mg-Cu alloy (AA7010) surfaces by etching technique was studied. The alloy surfaces were mechanically engraved with some identification marks using "Gravograph". The marks were then erased by removing the metal to different levels up to and below the depth of engraving. Five metallographic reagents were tested on the obliterated surfaces by etching. The following two methods (i) immersion in 10% aq. phosphoric acid and (ii) alternate swabbing of 60% HCl and 40% NaOH were found to be quite effective to reveal the obliterated marks. These two procedures were also able to show effectively the marks obliterated by over-engraving and centre punching. Of the two techniques immersion in phosphoric acid provided more contrast. Interestingly, alternate swabbing of 60% HCl and 40% NaOH presented itself to be the common reagent for restoration on pure aluminium as well as its alloy surfaces. This is evident from our own current experiments and those of earlier researchers [G. Peeler, S. Gutowski, H. Wrobel, G. Dower, The restoration of impressed characters on aluminium alloy motor cycle frames, J. Forensic Ident. 58 (1) (2008) 27-32; M. Izhar M. Baharum, R. Kuppuswamy, A.A. Rahman, Restoration of engraved marks on aluminium surfaces by etching technique, Forensic Sci. Int. 177 (2008) 221-227]. The findings have assumed importance as engines and chassis of cars and frames of firearms are currently made of high strength aluminium alloys and recovery on these surfaces by current methods is not satisfactory. PMID- 20022188 TI - Autonomic function and change in insulin for exercising postmenopausal women. AB - PURPOSE: Obesity, physical inactivity and altered estrogen metabolism play an integrated role contributing to the disease risk profiles of postmenopausal women. These same risk factors also affect modulation of the autonomic nervous system (ANS). METHODS: We examined 332 postmenopausal, overweight, previously sedentary women (mean+/-SD; age, 57.6+/-6.3 years; weight, 84.3+/-11.9kg; BMI, 31.7+/-3.7kg/m(2)) participating in a 6-month, moderate intensity, aerobic exercise training intervention to determine the relationship between heart rate variability (HRV) derived autonomic function and fasting insulin. We analyzed quartiles of change in time and frequency domain indices of ANS activity and changes in insulin for between and within group differences using ANCOVA and Tukey post hoc tests adjusted for age, ethnicity, randomization group, change in fitness, and change in weight. RESULTS: We observed at baseline that insulin was positively correlated with body anthropometry (body weight, r(2)=0.34; BMI, r(2)=0.39; waist circumference, r(2)=0.29; all, P<0.001), and inversely associated with rMSSD (r(2)=-0.12) and SDNN (r(2)=-0.18; all, P<0.01). After the intervention, changes in rMSSD (r(2)=-0.21, P<0.002) and SDNN r(2) -0.19, P<0.0001) were inversely correlated to insulin change. Further ANCOVA analysis revealed that rMSSD and SDNN were both significant (P<0.0001); however, only rMSSD exhibited a step-wise pattern of improvement when quartiles of rMSSD were compared to corresponding insulin reductions: Q1 (referent group, 8.41+/ 3.2uIU/ml), Q2 (-3.30+/--3.2uIU/ml), Q3 (-5.66+/--3.2uIU/ml; P<0.02), and Q4 ( 9.60+/--3.2uIU/ml; P<0.006). CONCLUSION: Our study shows that changes in autonomic function are associated with changes in insulin and that exercise training may influence this relationship in postmenopausal women. PMID- 20022190 TI - Brain metabolite changes in alcoholism: localized proton magnetic resonance spectroscopy study of the occipital lobe. AB - Chronic alcoholism is associated with altered brain metabolism, morphology and cognitive abilities. Besides deficits in higher order cognitive functions, alcoholics also show a deficit in the processing of basic sensory information viz. visual stimulation. To assess the metabolic changes associated with this deficit, (1)H MRS was carried out in the occipital lobe of alcohol dependents. A significant increase in Cho/Cr ratio (p<0.015) was observed in occipital lobe in the alcoholic group indicating altered cell membrane metabolism, which may probably be associated with the alterations in the cognitive abilities associated with vision. PMID- 20022191 TI - Virtual bronchoscopy-guided transbronchial biopsy for aiding the diagnosis of peripheral lung cancer. AB - OBJECTIVE: The aim of this study was to evaluate the clinical value of virtual bronchoscopy (VB) in aiding diagnosis of peripheral lung cancer by transbronchial biopsy (TBB). In addition, we sought to systematically analyze the factors that affect the diagnostic sensitivity of VB-guided TBB for the evaluation of peripheral lung cancers. MATERIALS AND METHODS: A hundred and twenty-two peripheral lung cancers from 122 patients (82 men and 40 women, 38-84 years; median 68.5 years) who were performed VB-guided TBB were evaluated retrospectively. VB was reconstructed from 1- or 0.5-mm slice thickness images of multi-detector CT (MDCT). Experienced pulmonologists inserted the conventional and ultrathin bronchoscopes into the target bronchus under direct vision following the VB image. RESULTS: A definitive diagnosis was established by VB guided TBB in 96 lesions (79%). The diagnostic sensitivity of small pulmonary lesions <=30 mm in maximal diameter (71%) was significantly lower than that of lesions >30 mm (91%, p=0.008). For small pulmonary lesions <=30 mm (n=76), internal opacity of the lesion was the independent predictor of diagnostic sensitivity by VB-guided TBB, and the non-solid type lung cancers were significantly lower than the solid type and part-solid type lung cancers for diagnostic sensitivity (odds ratio=0.161; 95% confidence interval=0.033-0.780; p=0.023). CONCLUSION: Use of an ultrathin bronchoscope and simulation with VB reconstructed by high quality MDCT images is thought to improve pathological diagnosis of peripheral lung cancers, especially for solid and partly solid types. For small pulmonary lesions <=30 mm, the lesion internal opacity is a significant factor for predicting the diagnostic sensitivity, and the sensitivity was low for small non-solid type of lung cancers. PMID- 20022192 TI - CEM-101, a novel fluoroketolide: antimicrobial activity against a diverse collection of Gram-positive and Gram-negative bacteria. AB - CEM-101 is a novel fluoroketolide with reported high potency against diverse groups of Gram-positive (Micrococcus spp., viridans group streptococci, Corynebacterium spp. Listeria monocytogenes, Clostridium spp., etc.) and Gram negative bacteria (Neisseria gonorrhoeae, Campylobacter jejuni, Helicobacter pylori, Bacteroides fragilis, Shigella spp., etc.), including mycoplasma and ureaplasma, as well as bacteria commonly associated with community-acquired respiratory tract infections and skin and skin structure infections. In this study, CEM-101 and comparator antimicrobials were tested against a collection of very low prevalence aerobic and anaerobic bacteria collected via the SENTRY Antimicrobial Surveillance Program platform. CEM-101 was highly active against all Gram-positive organisms (MIC(50), 0.015 microg/mL) as compared with telithromycin (MIC(50), 0.06 microg/mL), clarithromycin (MIC(50), 0.12 microg/mL), and erythromycin (MIC(50), 0.25 microg/mL). Among Gram-negative pathogens, CEM-101 also displayed a high potency against most strains (MIC(50), 4 microg/mL) but was found to be equivalent or less active when compared with other antimicrobials tested with MIC(50) values ranging from < or =0.12 microg/mL for levofloxacin to 8 microg/mL for telithromycin. Among the strict anaerobic species, CEM-101 activity mirrored that of the aerobic species: high activity against the Gram-positive anaerobes (MIC(50) results ranging from < or =0.03 microg/mL to 0.12 microg/mL) and equivalent or less susceptible against Gram negative anaerobes. Our in vitro antimicrobial susceptibility results for CEM-101 demonstrate better activity compared with other MLS(B) class agents among a diverse group of uncommonly isolated bacterial pathogens; these results provide an impetus for possible expanded indications during Phase 2 and 3 clinical trials. PMID- 20022193 TI - Susceptibility to tigecycline of isolates from samples collected in hospitalized patients with secondary peritonitis undergoing surgery. AB - Activity of tigecycline against nosocomial secondary peritonitis isolates collected along 18 months in 29 Spanish hospitals was tested by Etest in a central laboratory, considering Food and Drug Administration (FDA)/British Society for Antimicrobial Chemotherapy (BSAC)/European Committee on Antimicrobial Susceptibility Testing (EUCAST) breakpoints. A total of 600 facultative/aerobic isolates (392 Gram negative, 208 Gram positive) and 100 anaerobes were tested. None of the 220 Escherichia coli isolates was resistant to tigecycline (MIC(50)/MIC(90) = 0.25/0.5 microg/mL), with 0.5% (FDA breakpoint) and 3.6% (BSAC/EUCAST breakpoint) intermediate strains. All Extended-spectrum beta lactamase (ESBL)-producing E. coli isolates (15 strains), all Klebsiella pneumoniae, and Klebsiella oxytoca isolates (42 strains) were susceptible to tigecycline. No isolates resistant to tigecycline were found among Streptococcus viridans, Staphylococcus aureus, and Enterococcus faecium, but 18.9% of Enterococcus faecalis strains were intermediate following BSAC/EUCAST breakpoints. All (but 1) isolates of the Bacteroides fragilis group (n = 45) were tigecycline susceptible, as well as Gram-positive anaerobes. Tigecycline offers an adequate activity profile against isolates from secondary peritonitis when tested by Etest regardless of the breakpoints used for categorization. PMID- 20022194 TI - Mutations in alpha-actinin-2 cause hypertrophic cardiomyopathy: a genome-wide analysis. AB - OBJECTIVES: This study describes a genome-wide linkage analysis of a large family with clinically heterogeneous hypertrophic cardiomyopathy (HCM). BACKGROUND: Familial HCM is a disorder characterized by genetic heterogeneity. In as many as 50% of HCM cases, the genetic cause remains unknown, suggesting that other genes may be involved. METHODS: Clinical evaluation, including clinical history, physical examination, electrocardiography, and 2-dimensional echocardiography, was performed, and blood was collected from family members (n = 23) for deoxyribonucleic acid analysis. The family was genotyped with markers from the 10 cM AB PRISM Human Linkage mapping set (Applied Biosystems, Foster City, California), and 2-point linkage analysis was performed. RESULTS: Affected family members showed marked clinical diversity, ranging from asymptomatic individuals to those with syncope, heart failure, and premature sudden death. The disease locus for this family was mapped to chromosome 1q42.2-q43, near the marker D1S2850 (logarithm of odds ratio = 2.82, theta = 0). A missense mutation, Ala119Thr, in the alpha-actinin-2 (ACTN2) gene was identified that segregated with disease in the family. An additional 297 HCM probands were screened for mutations in the ACTN2 gene using high-resolution melt analysis. Three causative ACTN2 mutations, Thr495Met, Glu583Ala, and Glu628Gly, were identified in an additional 4 families (total 1.7%) with HCM. CONCLUSIONS: This is the first genome-wide linkage analysis that shows mutations in ACTN2 cause HCM. Mutations in genes encoding Z-disk proteins account for a small but significant proportion of genotyped HCM families. PMID- 20022195 TI - Antiemetic therapy for nausea and vomiting in the emergency department. AB - BACKGROUND: Antiemetic agents are among the most frequently prescribed medications in the emergency department (ED). Nevertheless, there are no widely accepted evidence-based guidelines to optimize the use of these medications for nausea or vomiting in this setting. OBJECTIVE: The objective of this article is to briefly review the evidence supporting the use of antiemetic agents for the treatment of nausea or vomiting for adults in the ED, and to provide recommendations to help guide therapy. DISCUSSION: The antiemetic agents studied include droperidol, promethazine, prochlorperazine, metoclopramide, and ondansetron. Droperidol was commonly used in the past, and is more effective than prochlorperazine or metoclopramide, but due to the US Food and Drug Administration black box warning regarding the potential for QT prolongation with this drug, its use is limited to refractory cases. Promethazine is more sedating than other comparative agents, and also has the potential for vascular damage upon intravenous administration. It may be a suitable option when sedation is desirable. Patients given prochlorperazine or metoclopramide must be monitored for akathisia that can develop at any time over 48 h post administration. Decreasing the infusion rate can reduce the incidence of this adverse effect, and the effect can be treated with intravenous diphenhydramine. Ondansetron is as effective as promethazine, and is not associated with sedation or akathisia. CONCLUSION: Based on the safety and efficacy of ondansetron, it may be used as a first-line agent for relief of nausea or vomiting for most patient populations in the ED. PMID- 20022196 TI - Lewis lead enhances atrial activity detection in wide QRS tachycardia. AB - BACKGROUND: The differential diagnosis of wide QRS tachycardia is a challenge for the emergency physician. The major tool is the electrocardiogram (ECG), even though the sensitivity and specificity may be variable, depending on presentation. Additional leads could be used to improve the diagnostic accuracy of the ECG. OBJECTIVE: To document the use of the Lewis lead in improving the diagnostic accuracy of the ECG in wide QRS tachycardia. CASE REPORT: A 52-year old woman with rheumatoid arthritis, in treatment with methotrexate, was admitted with progressive dyspnea that evolved to acute respiratory distress and shock at arrival. Pneumonia was diagnosed as the infection and she received antibiotics, and respiratory and inotropic support. She was also using amiodarone for more than 10 years, but she couldn't state the reason. On cardiac monitoring, wide QRS tachycardia was detected and ventricular tachycardia was considered on the differential diagnosis. The standard 12-lead ECG was complemented with the Lewis lead, obtained with higher speed and amplitude, demonstrating atrioventricular concordance and excluding ventricular tachycardia. The patient was treated for septic shock, and she died 2 days later. CONCLUSION: The Lewis lead is a simple and easy strategy to enhance atrial activity detection in wide QRS tachycardia. PMID- 20022197 TI - Two cases of penetrating abdominal injury from needlefish impalement. AB - BACKGROUND: Needlefish injuries to humans are extremely rare. An extensive review of the medical literature reveals only 22 cases previously reported. Most injuries reported have occurred in the Indo-Pacific region, but there are scattered reports worldwide. OBJECTIVE: To report two cases of life-threatening penetrating abdominal injuries from needlefish impalement. CASE REPORT: A 19-year old man and a 40-year-old woman were both struck in the abdomen by a needlefish and required emergent exploratory laparotomy for instability. Their injuries were a laceration of a branch of the left portal vein and a small bowel perforation, respectively. Both patients survived and recovered without incident. CONCLUSION: Although rare, penetrating injuries from needlefish can be life threatening. Treatment should be directed at the organ injured with appropriate antibiotic coverage. PMID- 20022198 TI - The rapid impact on mortality rates of a dedicated care team including trauma and emergency physicians at an academic medical center. AB - BACKGROUND: Trauma center designation can result in improved patient outcomes after injuries. Whereas the presence of trauma teams has been associated with improved trauma patient outcomes, the specific components, including the role of emergency medicine (EM)-trained, board-certified emergency physicians, have not been defined. OBJECTIVE: To assess the outcomes of patients before and after the establishment of a dedicated trauma team that incorporated full-time EM-trained physicians with trauma specialists at a Level I trauma center at an academic institution. METHODS: Secondary analysis of prospectively collected trauma registry data was performed to compare mortality rates of all treated trauma patients before and after this intervention. RESULTS: The establishment of a dedicated specialty trauma team incorporating full-time EM presence including EM trained, board-certified emergency physicians was associated with a reduction in overall non-DOA (dead on arrival) mortality rate from 6.0% to 4.1% from the time period preceding (1999-2000) to the time period after (2002-2003) this intervention (1.9% absolute reduction in mortality, 95% confidence interval [CI] 0.7%-3.0%). Among patients who were most severely injured (Injury Severity Score [ISS] >= 25), mortality rates decreased from 30.2% to 22.0% (8.3% absolute reduction in mortality, 95% CI 2.1%-14.4%). In comparison, there was minimal change in national mortality rates for patients with ISS >= 25 during the same time period (33% to 34%). CONCLUSIONS: The implementation of a dedicated full time trauma team incorporating both trauma surgeons and EM-trained, board certified or -eligible emergency physicians was associated with improved mortality rates in trauma patients treated at a Level I academic medical center, including those patients presenting with the most severe injuries. PMID- 20022199 TI - Influence of implementation of a chest pain unit on acute coronary syndrome outcomes. AB - BACKGROUND: Different strategies have been described to increase promptness and efficiency in the assessment and management of patients with acute chest pain and acute coronary syndrome (ACS) in the emergency department (ED). OBJECTIVE: The objective of this study is to evaluate the results of implementing a Chest Pain Unit (CPU) to assist patients with ACS, and to determine its impact on quality of health care indexes and clinical outcomes. METHODS: A study was conducted with a prospective cohort of patients admitted to the ED with a chief complaint of acute chest pain or suspected ACS at two different time periods: before (n = 663) and after (n = 450) introducing a CPU as part of the ED. Quality-of-care indexes analyzed in this study were adherence to a critical pathway, length of hospital stay, and hospital mortality. RESULTS: There was increased adherence to a critical pathway during the CPU period compared to the period with no designated CPU area, including compliance with prescribing aspirin, beta-blockers, and angiotensin-converting enzyme inhibitor, and performing coronary angiography in high-risk patients. After adjustment to baseline characteristics, admissions to a CPU resulted in a 65% reduction in mortality (odds ratio 0.35; 95% confidence interval 0.14-0.88; p = 0.03). There was no difference in median length of hospital stay, 7 days (interquartile range [IQR] 4-12) before CPU and 6 days (IQR 4-11) after introducing the CPU (p = 0.10). CONCLUSION: In the scenario of a crowded ED, implementation of a CPU was associated with greater adherence to a critical pathway for patients with ACS, with a concomitant reduction in mortality rates. PMID- 20022200 TI - Pneumoretroperitoneum as the manifestation of rectal perforation. PMID- 20022201 TI - Randomised controlled trial of a pedometer-based telephone intervention to increase physical activity among cardiac patients not attending cardiac rehabilitation. AB - OBJECTIVE: To determine the effectiveness of a pedometer-based telephone intervention on the physical activity levels of cardiac patients who did not attend a CRP. METHODS: A randomised controlled trial was conducted with 215 patients referred to a CRP but who could not or chose not to attend. The 6-week intervention included self-monitoring of physical activity using a pedometer and step calendar; and behavioural counselling and goal setting sessions. Data were collected at baseline, 6 weeks and 6 months. RESULTS: Study groups did not differ significantly at baseline. After 6 weeks, improvements in total physical activity time (p=0.027), total physical activity sessions (p=0.003), walking time (p=0.013) and walking sessions (p=0.002) in the intervention group were significantly greater than the control group after adjusting for baseline differences, and remained significant at 6 months. CONCLUSION: The findings that the pedometer-based telephone intervention was successful in increasing physical activity levels in cardiac patients who did not attend a CRP could result in major health benefits for this group of people. PRACTICAL IMPLICATIONS: The pedometer-based telephone intervention could be offered as an effective and accessible option for patients not attending a CRP to increase and maintain their physical activity levels after hospitalisation. PMID- 20022202 TI - Predictors of employment in substance-using male and female welfare recipients. AB - This study examined predictors of employment among substance-using men and women enrolled in welfare-to-work programs. Participants were 394 welfare applicants assigned to either coordinated care management or usual care for treatment services and job training and followed up for 1 year to track employment outcomes. Common barriers to work were assessed at baseline in four key domains: disabilities, situational barriers, labor capital, and motivation. Results indicated substantial gender differences in the number and profile of work barriers. Among men, work experience and job motivation were the only significant predictors of employment; among women, multiple factors from each barrier domain predicted job acquisition even when controlling for all other significant predictors. Findings suggest that welfare-to-work programs should emphasize job training and job seeking during the early stages of welfare interventions for men and for many women. PMID- 20022204 TI - Analysis of gender-related differences in lower extremity peripheral arterial disease. AB - INTRODUCTION: Gender-related differences continue to challenge the management of lower extremity (LE) peripheral arterial disease (PAD) in women. We analyzed the time-trends in hospital care of such differences. METHODS: Data for patients with PAD from New York, New Jersey, and Florida state hospital inpatient discharge databases (1998-2007) were analyzed using univariate and multivariate regression analyses. RESULTS: The 2.4 million PAD-related inpatient discharge records analyzed showed a slight decrease of inpatient procedures for both genders. Compared with men, women had 18% to 27% fewer PAD and 33% to 49% fewer vascular procedural hospitalizations (P < .0001). They were persistently more likely than men to be admitted emergently (56% vs 51% in 1998 and 57% vs 53% in 2007) and discharged to a nursing home. During the study period, the amputation rate declined by 36% in women and 21% in men with PAD, and similarly, open procedures decreased by 36% and 30%. Endovascular procedures, however, increased by 150% in women and 144% in men. Procedural mortality was 4.95% vs 4.37% for men (P < .0001). Female mortality rates were persistently higher after amputations (9.89% vs 8.90%, P < .0001), open (5.49% vs 4.00%, P < .0001), and endovascular procedures (2.87% vs 2.10%, P < .0001). Time trends showed improved mortality for men and women, with a stable difference between the two. CONCLUSION: The analysis of representative state administrative databases of inpatient care records demonstrated improvements in mortality and amputation rates over time. However, a gender-related disparity in PAD outcomes remains that merits further investigation. PMID- 20022205 TI - A survey of demographics, motivations, and backgrounds among applicants to the integrated 0 + 5 vascular surgery residency. AB - OBJECTIVE: The 0 + 5 integrated vascular surgery (VS) residency has altered the training paradigm for future vascular specialists. Rising interest in these novel programs highlights our need to better understand the applicant pool. We compared demographics and surveyed recent applicants to our integrated program to gain more insight into their background and motivation for accelerated vascular training. METHODS: Demographics and objective parameters were determined from all 65 applicants to the integrated VS program at Stanford University Medical Center and compared to 58 applicants interviewed by the general surgery (GS) program at Harbor-UCLA Medical Center by querying the Electronic Residency Application System for the programs in 2009. There was no overlap of applicants between programs. An anonymous, voluntary Web-based survey was sent to these cohorts with a response rate of 82% for VS applicants and 60% for GS applicants. Subjects were queried regarding their background, personal experience, prior exposure to VS, and motivations for residency specialty selection. RESULTS: Applicants to integrated VS programs tended to be older, were less likely to be from a US medical school, had a higher number of publications, and a higher percentage of cardiovascular-related publications than the GS applicants. When stratified by the 27 VS applicants (41%) that were offered an interview, this highly selected and desirable group for training was nearly 40% female, more likely to have an additional degree (PhD, master's), just as likely to be in the top quartile of their medical school class (60%), and score equally well on standardized board examinations (90th percentile) than the top GS applicants offered interviews. Survey data revealed that the majority of career choices (65%) were made during the third and fourth years of medical school. Factors most strongly influencing the decision to choose VS as a career were endovascular technologies/devices, challenging open vascular operations, clinical rotations on vascular surgery, the aging patient population, and perceived need for vascular surgeons and vascular surgeon mentorship. The most common reasons cited for particularly pursuing an integrated 0 + 5 VS training program were (1) more focused training/integration of cardiovascular medicine, (2) interest in catheter-based endovascular therapies, and (3) shorter time in training. Of the GS applicants, 58% indicated they would be interested in applying to an integrated residency in their subspecialty of interest, and 45% listed vascular surgery as a potential fellowship option after general surgery. CONCLUSION: Applicants to 0 + 5 integrated vascular residencies were more likely to have rotated on a vascular surgery service, observed vascular cases, identified a vascular surgery mentor, and been actively involved in cardiovascular research. The quality of the top VS applicant based on class rank and test scores is comparable to the top GS applicants, yet the VS applicant has a higher percentage of advanced degrees, more publications, and more involvement in cardiovascular research. Institutional strategies to increase medical student exposure to vascular surgery clinically and via research programs will optimize our ability to attract and train the best candidates in these new training programs. PMID- 20022206 TI - Perioperative beta-blockers for vascular surgery patients. AB - Perioperative beta-blocker therapy has been a heavily investigated and controversial topic during the past decade. Prior national consensus statements that recommended the routine use of these medications in patients undergoing high risk surgical procedures have been called into question because of the results of recent clinical trials that involved heterogeneous groups of surgical patients. This article reviews the evidence for perioperative beta-blocker usage as it pertains to patients undergoing vascular surgery procedures. The weight of evidence suggests that beta-blockers lower the perioperative risk of myocardial ischemia or infarction and cardiovascular death among patients with clinical risk factors undergoing major vascular surgery. However, there appears to be a concurrent risk of adverse events associated with these medications if patients are not monitored properly during the perioperative period. Perioperative beta blockers should continue to occupy a prominent role in the therapeutic armamentarium for improving outcomes among high-risk patients undergoing major vascular surgery. PMID- 20022207 TI - Technical aspects and midterm patency of iliac branched devices. AB - OBJECTIVE: This study presents technical aspects and initial results with iliac bifurcated devices (IBDs). METHODS: Since 2006, 47 IBDs were scheduled for 37 patients who were followed up between 2 and 31 months. Iliac aneurysms were unilateral in 27 patients and bilateral in 10. Two patients with bilateral common iliac artery aneurysms (CIAAs) did not have a simultaneous aortic aneurysm. Two patients underwent combined thoracoabdominal aneurysm treatment with branched stent grafts, and one underwent combined juxtarenal aneurysm repair with a fenestrated device. The helical iliac side branch device was used in 11 CIAA (23.4%), and the Zenith bifurcated iliac side branch device was used in the remaining 36 (76.6%). RESULTS: The technical success rate was 97.3% within the 47 intended-to-treat CIAAs (failure to introduce the delivery system in one case, converted to femorofemoral bypass). During follow-up, five (10.6%) hypogastric branch occlusions occurred in five patients. Two patients with bilateral repair had unilateral internal iliac artery side branch occlusions without ischemic symptoms. In contrast, of the three patients with unilateral side branch occlusion and simultaneous contralateral internal iliac artery occlusion (2 chronic and 1 coil embolization), persistent buttock claudication and sexual dysfunction developed in one. The secondary patency, including one redo case, was 87.3% at 22 months (standard error <10%). CONCLUSIONS: The use of branched stent grafts is a feasible procedure, including for patients with bilateral iliac aneurysmal disease or concomitant juxtarenal or thoracoabdominal aortic disease. PMID- 20022208 TI - Impact of metabolic syndrome on the outcomes of percutaneous renal angioplasty and stenting. AB - BACKGROUND: Endovascular therapy for symptomatic atherosclerotic renal artery stenosis (ARAS) is common and effective in the well-selected patient. Hypertension is a common indication for intervention and a major component of metabolic syndrome (MetS). The impact of MetS on outcomes after percutaneous renal intervention is unknown. METHODS: We performed a retrospective analysis of records from patients who underwent endovascular intervention for ARAS and were followed by duplex ultrasound between January 1990 and January 2008. MetS was defined as the presence of >or=3 of the following criteria: Blood pressure >or=140 mm Hg/>or=90 mm Hg; triglycerides >or=150 mg/dL; high-density lipoprotein or=110 mg/dL; or body mass index >or=30 kg/m(2). The average follow-up period was 3.3 years. Clinical benefit defined as freedom from renal-related morbidity (increase in persistent creatinine >20% of baseline, progression to hemodialysis, death from renal-related causes) or freedom from recurrent hypertension, anatomic patency, restenosis, and patient survival were measured. RESULTS: Five hundred ninety-two renal artery interventions were performed in 427 patients. Fifty-two percent were identified as having MetS. Patients with MetS were more often female (35% vs 50%, NoMetS vs MetS). There were no significant differences in presenting symptoms. There was no peri-operative mortality and equivalent morbidity (6% vs 7%, NoMetS vs MetS). Patients with MetS had equivalent survival and cumulative patency. However, the MetS group had a lower five-year freedom from restenosis (87+/-2% vs 69+/-9%, NoMetS vs MetS; P < .01) and lower five-year retained clinical benefit (71+/-8% vs 45+/-8%, NoMetS vs MetS; P < .01) with a higher number progressing to hemodialysis (3% vs 13%, NoMetS vs MetS; P < .01). Individually, the components of MetS did not influence outcomes. Statin therapy did not influence outcomes. CONCLUSION: MetS is associated with markedly reduced renal clinical benefit and increased progression to hemodialysis following endovascular intervention for atherosclerotic renal artery stenosis. MetS is thus a risk factor for poor long-term outcomes following renal interventions. PMID- 20022209 TI - Prevention of thromboembolic events in surgical patients through the creation and implementation of a computerized risk assessment program. AB - OBJECTIVES: Deep vein thrombosis (DVT) is a major source of postoperative morbidity and mortality and is currently a major quality improvement initiative. Mechanical and pharmacological prophylaxis is effective in preventing postoperative thromboembolic events, yet it remains underutilized in the clinical setting. Thus, the objective of this study was to develop and implement a computerized DVT risk assessment program in the electronic medical record and determine its effect on compliance with DVT prophylaxis guidelines. METHODS: A standardized DVT risk assessment program was developed and incorporated into the Computerized Patient Record System for all surgical patients at the Jesse Brown Veterans Affairs Medical Center. Four hundred consecutive surgical patients before and after implementation were evaluated for DVT risk, the prescription of pharmacological and mechanical DVT prophylaxis, and the development of thromboembolic events. RESULTS: With implementation of the DVT risk assessment program, the number of patients receiving the recommended pharmacological prophylaxis preoperatively more than doubled (14% to 36%) (P < .001), and use of sequential compression devices (SCD) increased 40% (P < .001). Overall, the percentage of at-risk patients receiving the recommended combined DVT prophylaxis of SCD and pharmacological prophylaxis increased nearly seven-fold (5% to 32%) (P < .001). The assessment also improved use of prophylaxis postoperatively, increasing SCD use by 27% (P < .001). With respect to DVT occurrence, there was an 80% decrease in the incidence of postoperative DVT at 30 days and a 36% decrease at 90 days; however, this did not reach statistical significance due to the low event rate. CONCLUSIONS: The creation and implementation of a standardized DVT risk assessment program in the electronic medical record significantly increased use of pharmacological and mechanical DVT prophylaxis before surgery in a Veterans Affairs Medical Center setting. PMID- 20022210 TI - The anatomy of the small saphenous vein: fascial and neural relations, saphenofemoral junction, and valves. AB - PURPOSE: Varicose veins are a frequent burden, also in the small saphenous system. Yet its basic anatomy is not described consistently. We therefore investigated the fascial and neural relationships of the small saphenous vein (SSV) as well as the frequency and position of valves and the different junctional patterns, also considering the thigh extension. MATERIALS AND METHODS: We dissected the legs of 51 cadavers during the regular dissection course held in winter 2007 at Innsbruck Medical University, with a total of 86 SSVs investigable proximally and 94 SSVs distally. RESULTS: A distinct saphenous fascia is present in 93 of 94 cases. It starts with a mean distance of 5.1 cm (SD 1.2 cm) proximal to the calcaneal tuber, where the tributaries to the SSV join to form a common trunk. The neural topography at the level of the gastrocnemius muscle's origins shows the medial sural cutaneous nerve in 88% medially and in 12% laterally to the SSV, the tibial nerve in 64% medially and in 36% laterally, and the common fibular nerve in 98% medially and in 2% laterally to the vein. The saphenopopliteal junction (SPJ) resembled in about 37% type A (UIP classification), 15% type B, and 24% type C. A total of 17% of specimens showed a venous web or star at the popliteal fossa and 6% had a doubled junction. A thigh extension could be demonstrated in about 84%. A most proximal valve was present in only 94% at a mean distance of 1.2 cm (SD 1.4 cm) to the SSVs orifice. A consecutive distal valve was only present in 65% with a mean distance of 5.1 cm (SD 2.3 cm). CONCLUSION: Two fascial points or regions can be described in the SSVs' course and its own saphenous fascia is demonstrated macroscopically in almost all cases. The neural topography is highly individual. The SPJ is highly individual where we found hitherto unclassified patterns in a remarkable number of veins. Venous valves are not as frequent as we supposed them to be. Furthermore, not all most proximal valves seem to be terminal valves. PMID- 20022212 TI - Hemin prevents in-stent stenosis in rat and rabbit models by inducing heme oxygenase-1. AB - OBJECTIVE: The introduction of drug-eluting stents (DES) has largely added benefit to the percutaneous coronary intervention. Questions about the long-term safety of DES have been raised, however, particularly with respect to late stent thrombosis. Research efforts are now being directed toward therapeutics that can impede smooth muscle proliferation and promote vascular healing. Emerging data suggest that heme oxygenase-1 (HO-1), an inducible oxidoreductase enzyme system, can exert cytoprotective effects on endothelial cells and limit smooth muscle cell proliferation. We assessed the ability of hemin, a potent HO-1 inducer, to reduce in-stent stenosis without compromising re-endothelialization. METHODS: Rat aorta and rabbit iliac arteries were stented. Animals received ongoing treated with intraperitoneal hemin (50 mg/kg) or vehicle. At 7 to 28 days after surgery, stented arterial segments were collected and processed for histologic, electron microscopy, or protein analysis. RESULTS: In both models, treatment with hemin reduced neointima growth without compromising re-endothelialization of the stented arteries. In the rat aorta, analysis of protein expression at 7 and 28 days after stenting revealed that hemin increased HO-1 expression and limited the early inflammatory, apoptotic, and proliferative cellular events that are common to in-stent stenosis. Hemin treatment decreased the expression of the Ki-67 protein and the activity of key regulators of smooth muscle cell proliferation, including p42/44, RhoA, and up-regulated the expression of cyclin-dependent kinase inhibitors. The beneficial effects of hemin were abolished in the presence of tin-protoporphyrin IX, an HO inhibitor. Finally, treatment with tricarbonylchloro(glycinato)ruthenium(II), a carbon monoxide donor, reduced in stent stenosis in the rat aorta, suggesting that carbon monoxide, a by-product of heme degradation, might contribute to the protective effect of hemin. CONCLUSION: These results suggest that HO-1 is important in limiting in-stent stenosis and can be regarded as a new therapeutic target. PMID- 20022213 TI - Dealing honestly with an honest mistake. AB - A 70-year-old woman was admitted for a symptomatic left iliofemoral deep vein thrombosis. She underwent percutaneous mechanical thrombectomy, followed by overnight thrombolysis. The next day her clot had resolved, and a culprit left iliac vein stenosis was identified. After stent placement, a heparin infusion was initiated and the patient was taken back to the ward. At 11 the evening after the procedure, the resident on call was contacted to verify the written order. The resident stated that the heparin dose was to be 250 U/h; however, the nurse documented 2500 U/h and changed the infusion pump at the patient's bedside. At 5:30 the next morning, the resident was notified that the patient's partial thromboplastin time was >300 seconds and promptly shut off the heparin infusion. No noticeable adverse events occurred because of the high heparin dosing. The charge nurse was notified, as was risk management. What should the patient be told? PMID- 20022215 TI - Statin use does not affect the outcome of acute infection: a prospective cohort study. AB - Statins are used to prevent atherosclerosis because of their hypolipemic effects. In vitro testing and murine models suggest that statins may affect outcome in sepsis. Our meta-analysis of epidemiological studies in humans confirms that previous statin use appears to have a protective effect on infection rates and outcomes. The studies considered, however, were very different and heterogeneity was high, especially for mortality criteria. OBJECTIVE: To compare outcome for current statin users and nonusers hospitalized with fever, under pragmatic circumstances. DESIGN: Prospective cohort study. SETTING: University Hospital of Brest (France). PARTICIPANTS: Febrile (>38 degrees C) patients older than 40 years, admitted to the hospital on an emergency basis in 2005. Patients' outcome was compared according to their exposure to statins (current user or nonuser). MAIN OUTCOME MEASURES: Mortality, length of hospitalization, admission to ICU and to convalescent homes RESULTS: Of 40 343 patients referred for admission by the emergency department in 2005, 964 patients older than 40 years had a fever higher than 38 degrees C and were included in the study. Statin-user status, however, was available for only 921. The ICU admission rate (relative risk: 4.69; 95% CI: 2.42-9.08) was significantly higher (p=0.009) among statin users (n=139) than nonusers (n=782). The groups had similar lengths of hospitalization and similar rates of mortality and of convalescent home admissions. CONCLUSION: Our study does not confirm that statin use affects infection outcome and suggests that previous findings may be associated instead with more cautious care or nonspecific cardiovascular prevention rather than with a specific infection related effect. Thus, these data support the continuation of statin therapy during physiological aggression, including infection. PMID- 20022217 TI - Augmentation of the atrophic edentulous mandible by a bilateral two-step osteotomy with autogenous bone graft to place osseointegrated dental implants. AB - Extensive resorption of the mandible increases the interarch space and rehabilitation with traditional dentures is often unsatisfactory due to the superficialization of intraoral muscles. A study of 19 patients who underwent augmentation of an atrophic mandible using a bilateral two-step osteotomy and interpositional bone graft technique is presented. Three horizontal bone cuts (one in the intraforamina and two in the molar region) were made and jointed together by two short vertical bone cuts mesialy to the mental nerve. The cranial fragment was lifted and the iliac bone graft was interposed recreating the correct intermaxillary relationship. A broad vascular pedicle was maintained during surgery, ensuring nutrition from the lingual side, essential to reduce resorption of the bone graft and cranial fragment. 141 Biomet 3i Osseotite((R)) implants were placed. Patients were rehabilitated with a full-arch implant supported fixed prosthesis or an implant-supported overdenture. This clinical study describes the resorption process over a 4 year follow-up. 3 of 19 suffered from persistent neurosensitive disturbances. In conclusion, bilateral two-step osteotomy in association with interpositional bone graft is a reliable surgical means to recreate the anatomical morphology of the mandible. PMID- 20022216 TI - Hepatitis C and interferon induced thyroiditis. AB - Autoimmune thyroid diseases (AITDs) are complex diseases that develop as a result of interactions between genetic, epigenetic, and environmental factors. Significant progress has been made in our understanding of the genetic and environmental triggers contributing to AITD. The major environmental triggers of AITD include iodine, smoking, medications, pregnancy, and possibly stress. In this review we will focus on two well-documented environmental triggers of AITD, hepatitis C virus (HCV) infection and interferon alpha (IFNa) therapy. Chronic HCV infection has been shown to be associated with increased incidence of clinical and subclinical autoimmune thyroiditis (i.e. the presence of thyroid antibodies in euthyroid subjects). Moreover, IFNa therapy of chronic HCV infection is associated with subclinical or clinical thyroiditis in up to 40% of cases which can be autoimmune, or non-autoimmune thyroiditis. In some cases interferon induced thyroiditis (IIT) in chronic HCV patients may result in severe symptomatology necessitating discontinuation of therapy. While the epidemiology and clinical presentation of HCV and interferon induced thyroiditis have been well characterized, the mechanisms causing these conditions are still poorly understood. PMID- 20022218 TI - Smoking initiation and schizophrenia: a replication study in a Chinese Han population. AB - Schizophrenia is associated with a greater probability of ever smoking daily and with higher rates of initiation of daily smoking after age 20 in Caucasian populations. The aims of the current study were to replicate that schizophrenia is associated with smoking and higher risk of initiating daily smoking before schizophrenia starts among a large sample of male Chinese patients. A survival analysis of onset age for daily smoking compared 776 DSM-IV male inpatients with schizophrenia to 560 male controls. The results showed that the cumulative hazard curves for age of smoking initiation in schizophrenia and controls were significantly different (p<0.001), even after controlling for education (p<0.001). After excluding the patients who started smoking within 5 years before schizophrenia started, the cumulative hazard curve for schizophrenia was significantly different from ever-smoked controls (p<0.001), even after adjusting for education (p<0.001). These findings suggest that schizophrenic patients have a higher risk of starting daily smoking suggesting that vulnerability to schizophrenia may be associated with a higher risk of becoming a daily smoker. PMID- 20022219 TI - Impaired decision making in schizophrenia and orbitofrontal cortex lesion patients. AB - BACKGROUND: The aim of this study was to examine impaired decision making in patients with schizophrenia and in patients with orbitofrontal cortex lesions. METHODS: Schizophrenia patients (N=21), healthy controls (N=20) and an independent group of orbitofrontal patients (N=10) underwent a computerized version of the "Regret Gambling Task". Participants chose between two gambles, each having different probabilities and different expected monetary outcomes, and rated their emotional states after seeing the obtained outcome. Regret was induced by providing information about the outcome of the unchosen gamble. RESULTS: Healthy controls reported emotional responses consistent with counterfactual reasoning between obtained and unobtained outcomes; they chose minimizing future regret and were able to learn from their emotional experience. In contrast, orbitofrontal patients and schizophrenia patients with prominent positive symptoms did not report any regret and did not anticipate any negative consequences of their choices. Our results demonstrate first the presence of very different behavioural deficits within the spectrum of schizophrenia patients which may have contributed to the discrepancies observed in previous studies. Second, the results suggest that a subgroup of schizophrenia patients might have an orbitofrontal dysfunction, in fact, schizophrenia patients with positive symptoms have a behavioural dysfunction analogous to that of the orbitofrontal patients. CONCLUSION: Schizophrenia patients with prominent positive symptoms were unable to integrate cognitive and emotional components of decision making which may contribute to their inability to generate adaptive behaviours in social and individual environments. PMID- 20022220 TI - Inclusion of OXA-143 primers in a multiplex polymerase chain reaction (PCR) for genes encoding prevalent OXA carbapenemases in Acinetobacter spp. PMID- 20022221 TI - Detection of bla(CTX-M-15) extended-spectrum beta-lactamase genes in Escherichia coli from hospital patients in Nigeria. PMID- 20022222 TI - Region- and phase-dependent effects of 5-HT(1A) and 5-HT(2C) receptor activation on adult neurogenesis. AB - Adult neurogenesis and serotoninergic transmission are associated to mood disorders and their treatments. The present study focused on the effects of chronic activation of 5-HT(1A) and 5-HT(2C) receptors on newborn cell survival in the dentate gyrus (DG) and olfactory bulb (OB), and examined whether potential neurogenic zones as the prefrontal cortex (PFC) and striatum (ST) are reactive to these treatments. Administration of 8-OH-DPAT, but not RO600,175 increases neurogenesis and survival of late differentiating cells (15-21days) in the DG. Both 8-OH-DPAT and RO600,175 increase neurogenesis in the OB, but only 8-OH-DPAT affected cell survival, inducing a parallel decrease in the number of BrdU cells in the OB and increase in the SVZ, which suggests an impaired migration. In the PFC and ST, 8-OH-DPAT and R0600,175 increase gliogenesis (NG2-labeled cells). This study provides new insights on the serotonergic regulation of critical phases of neurogenesis helpful to understand the neurogenic and gliogenic effects of antidepressant treatments in different brain regions. PMID- 20022223 TI - No interactions between genetic polymorphisms and stressful life events on outcome of antidepressant treatment. AB - Genetic polymorphisms seem to influence the response on antidepressant treatment and moderate the impact of stress on depression. The present study aimed to assess, whether allelic variants and stressful life events interact on the clinical outcome of depression. In a sample of 290 systematically recruited patients diagnosed with a single depressive episode according to ICD-10, we assessed the outcome of antidepressant treatment and the presence of stressful life events in a 6-month period preceding onset of depression by means of structured interviews. Further, we genotyped nine polymorphisms in the genes encoding the serotonin transporter, brain derived neurotrophic factor, catechol-O methyltransferase, angiotensin converting enzyme, tryptophan hydroxylase, and the serotonin receptors 1A, 2A, and 2C. We found no evidence that the effects of the genetic polymorphisms on treatment outcome were dependent on stressful life events experienced by the individual prior to onset of depression. PMID- 20022224 TI - The influence of liposomal formulation factors on the interactions between liposomes and hydroxyapatite. AB - Liposomes may have potentials as a drug delivery system in the oral cavity; hence, the adsorption to, oral tissues may be of importance. The aim of this study was to find an optimal liposomal formulation with appropriate in vitro stability and which liposomal formulation parameters may be of importance for the interaction to tooth enamel surfaces. Charged liposomes were adsorbed in vitro onto hydroxyapatite (HA), used as a model substance for human dental enamel. For a systematic approach of lipid selection, statistical experimental design and multivariate analysis were conducted to interpret the data. The factors investigated were the type of charge (positive, negative), type of main phospholipid (egg-PC, DPPC, DMPC), type of charged lipid (diacyl-TAP, -ethylPC, PA, -PG, -PS), the amount of charged component (2.5, 10mol%) and the inclusion of cholesterol in the lipid bilayer. The results indicated that positively charged liposomes expressed significantly higher adsorption levels than the negatively charged ones. The effect of incorporating cholesterol did not turn out to be significant. Both positive egg-PC and DPPC liposomes exhibited high adsorption levels; however egg-PC liposomes were unstable during storage. For positively charged liposomes, the factor "type of main lipid" was found to be of significance for the adsorption, whereas, for negatively charged liposomes, no such important factors were found. Based on the adsorption profile to HA and the in vitro stability in phosphate buffer, the most promising liposomal formulation to target for human enamel in this study was the positively charged DPPC liposomes with 10mol% charged lipid included. However, more experiments are needed to determine the optimum mol% of positively charged lipid for the adsorption onto HA. PMID- 20022225 TI - Detection of DNA damage induced by styrene oxide in dsDNA layer-by-layer films using adriamycin as electroactive probe. AB - The detection of DNA damage is one of the most important topics in the DNA research fields. In this paper, an electrochemical method for the detection of DNA damage by combining the layer-by-layer assembly film with adriamycin (ADM) as an electrochemical probe was developed. Firstly, the layer-by-layer {dsDNA/PEI}(n) film was prepared by the alternate adsorption of polycationic polyethyleneimine (PEI) and negatively charged natural DNA onto glassy carbon electrode (GCE) surface. Its electrochemical behaviors were characterized by electrochemical impedance spectroscopy (EIS) and differential pulse voltammetry (DPV), and the optimal value of n was determined to be 2. Secondly, the {dsDNA/PEI}(2) film was immersed into the solution of ADM and the {dsDNA/PEI}(2) ADM was obtained. While transferred into the blank solution, the {dsDNA/PEI}(2) ADM would gradually release ADM, exhibiting the good reversibility of ADM incorporation. Finally, the DNA damage induced by styrene oxide (SO) was investigated and the promising results showed that the present method can be a useful tool for the detection of DNA damage. PMID- 20022226 TI - Effect of surface modification on the in vitro calcium phosphate growth on the surface of poly(methyl methacrylate) and bioactivity. AB - Poly(methyl methacrylate) (PMMA) is a biocompatible polymer widely used for bone substitutes. Its surface properties, however, are not favorable for the induction of biological apatite which can be directly related to natural bone formation. In this study, the surface of PMMA was modified by NaOH treatment or sequential treatments with ethanol (EtOH) and NaOH. Results displayed that surface hydrophilicity was improved for increasing treatment time and NaOH concentration. Field-emission scanning electron microscope (FE-SEM) displayed that in vitro formation of calcium phosphate (CaP) coating was significantly promoted by the surface modifications. X-ray photon spectroscopy (XPS) examination elucidated that the films prepared on PMMA consisted of calcium and phosphorus and their values for Ca/P ratio were closed to octacalcium phosphate (OCP). Fourier transform infrared (FT-IR) spectra of the film coated on PMMA revealed a band characteristic of phosphate groups confirming that CaP films were formed and their characteristics were dependent on the surface properties of PMMA. Cellular assay demonstrated that the adhesion of osteoblast-like MG63 cells was significantly promoted on CaP-coated PMMA. Proliferation assay showed that CaP films appeared not to exert any cytotoxic effects on the growth of MG63 cells. PMID- 20022227 TI - Oxidation of tyrosine by permanganate in presence of cetyltrimethylammonium bromide. AB - In this paper we report the effect of cetyltrimethylammonium bromide, CTAB on the oxidative degradation of tyrosine by permanganate. The reaction rate bears a first-order dependence on the [MnO(4)(-)] under pseudo-first-order conditions (large excess of [tyrosine] for at least 10 times over [MnO(4)(-)]) in presence of 10.0x10(-4)moldm(-3) CTAB. The effect of total [CTAB] on the reaction rate was determined. When [CTAB] was less than its critical micelle concentration (CMC) the rate constants (k(psi)) values decreased from 18.5x10(-4) to 7.2x10(-4)s(-1). As the [CTAB] was greater than the CMC, the k(psi) values increase from 7.2x10( 4) to 15.8x10(-4)s(-1)at room temperature. The premicellar environment of CTAB strongly inhibits the reaction rate where as increase in rate constant ascribed to the incorporation of tyrosine and MnO(4)(-) in to the Stern layer of CTAB micelles. The reaction has acid-dependent and acid-independent paths and, in the former case, the zero-order kinetics with respect to [H(2)SO(4)] shifted to fractional-order at higher [H(2)SO(4)]. Experiments have been done to confirm the nature of Mn(IV) formed during the reduction of permanganate by tyrosine. The mechanism with the observed kinetics has been proposed and discussed. The presence of -OH group is responsible for the higher reactivity of tyrosine which easily transfers the proton to MnO(4)(-). PMID- 20022229 TI - Serum paraoxonase 1 activity and oxidant/antioxidant status in Saudi women with polycystic ovary syndrome. AB - Oxidative stress is considered to be implicated in the pathophysiology of polycystic ovary syndrome (PCOS). This study was designed to evaluate the paraoxonase 1 (PON1) activity and oxidant/antioxidant status in Saudi women with PCOS and its contribution to the risk of atherosclerosis. Lipid profile, hormonal parameters, serum PON1 activity and oxidant (malondialdehyde)/antioxidant (total antioxidant capacity (TAC) levels were analyzed in 35 patients with PCOS and 30 healthy controls using a spectrophotometric method; correlation analysis was made between these variables. Insulin resistance was calculated by homeostasis model assessment (HOMA-IR). Women with PCOS had significantly higher fasting insulin, HOMA-IR and LH levels than controls. Lipid profiles and free androgen index (FAI) were significantly higher in women with PCOS when compared with controls. Serum PON1 activity was lower in the PCOS group (161.2+/-6.1U/l vs. 217.6+/-9.3U/l, p<0.001) compared with controls, whereas malondialdehyde levels were higher in the PCOS group (4.26+/-0.18nmol/ml vs. 1.37+/-0.12nmol/ml, p<0.001) compared with controls. Total antioxidant capacity was lower in the PCOS group (0.88+/ 0.10mmolTrolox/l vs. 1.63+/-0.17mmolTrolox/l, p<0.001) compared with controls. In PCOS group, serum PON1 was positively correlated with HDL-C (r=0.425, p<0.05) and TAC (r=0.582, p<0.01) but inversely correlated with HOMA-R (r=-0.54, p<0.01), testosterone (r=-0.672, p<0.01), FAI (r=-0.546, p<0.01) and malondialdehyde (r= 0.610, p<0.01). In conclusion, our data indicate that PON1 activity and antioxidant status were significantly decreased in Saudi women with PCOS. Lower serum PON1 activity might contribute to the increased susceptibility for the development of atherosclerosis risk in Saudi women with PCOS. Therefore, measurement of serum PON1 activity may be of value in assessment of women at higher risk for development of atherosclerosis risk in PCOS. However, further studies with larger sample size are needed to verify these results, and to assess the efficacy of antioxidant therapy on these patients. PMID- 20022230 TI - [Prolonged electrocerebral depression after oral administration of pyridoxine for pyridoxine-dependent convulsions]. PMID- 20022231 TI - Sensitive and specific detection of phaseolotoxigenic and nontoxigenic strains of Pseudomonas syringae pv. phaseolicola by TaqMan real-time PCR using site-specific recombinase gene sequences. AB - Pseudomonas syringae pv. phaseolicola, the causative agent of halo blight, is the most important bacterial pathogen of bean. Both nontoxigenic (Tox(-)) and toxigenic (Tox+) strains of this pathogen cause halo blight in beans. However, nontoxigenic strains cannot be detected by currently available molecular and serological tools. In this study, a TaqMan probe and primer set were designed based on the phage integrase family site-specific recombinase of P. s. pv. phaseolicola 1448A because it is known that most site-specific recombinases are structurally and functionally diverse. The specificity of the probe and primers was evaluated using purified DNA from 29 isolates of 3 different pathovars of P. syringae. The probe and primer set were able to detect Tox(-) and Tox+ isolates of P. s. pv. Phaseolicola, but no other phytopathogenic bacteria. The assay was also able to detect at least 5 genome equivalents of cloned amplified target DNA, using purified DNA, or 7 colony forming unit (CFU) per reaction when using calibrated cell suspensions. Thus, the TaqMan real-time PCR-based method can be used for the rapid detection of both types of P. s. pv. Phaseolicola, and will potentially simplify and facilitate the diagnosis and monitoring of this pathogen, and guide plant disease management. PMID- 20022232 TI - Discovery of the mitotic selective chromatid segregation phenomenon and its implications for vertebrate development. AB - The asymmetric cell division process is required for cellular differentiation and embryonic development. Recent evidence obtained in Drosophila and C. elegans suggest that this process occurs by non-equivalent distribution of proteins or mRNA (intrinsic factors) to daughter cells, or by their differential exposure to cell extrinsic factors. In contrast, haploid fission yeast sister cells developmentally differ by inheriting sister chromatids that are differentiated by epigenetic means. Specifically, the act of DNA replication at the mating-type locus in yeast switches it's alternate alleles only in one specific member of chromosome 2 sister chromatids in nearly every chromosome replication cycle. To employ this kind of mechanism for cellular differentiation, strictly based on Watson-Crick structure of DNA in diploid organism, selective segregation mechanism is required to coordinate distribution of potentially differentiated sister chromatids to daughter cells. Genetic evidence to this postulate was fortuitously provided by the analysis of mitotic recombinants of chromosome 7 in mouse cells. Remarkably, the biased segregation occurs in some cell types but not in others and the process seems to be chromosome-specific. This review summarizes the discovery of selective chromatid segregation phenomenon and it suggests that such a process of Somatic Sister chromatid Imprinting and Selective chromatid Segregation (SSIS model) might explain development in eukaryotes, such as that of the body axis left-right visceral organs laterality specification in mice. PMID- 20022228 TI - Gastrointestinal lymphatics in health and disease. AB - Lymphatics perform essential transport and immune regulatory functions to maintain homeostasis in the gastrointestinal (GI) system. Although blood and lymphatic vessels function as parallel and integrated systems, our understanding of lymphatic structure, regulation and functioning lags far behind that of the blood vascular system. This chapter reviews lymphatic flow, differences in lymphangiogenic and hemangiogenic factors, lymphatic fate determinants and structural features, and examines how altered molecular signaling influences lymphatic function in organs of the GI system. Innate errors in lymphatic development frequently disturb GI functioning and physiology. Expansion of lymphatics, a prominent feature of GI inflammation, may also play an important role in tissue restitution following injury. Destruction or dysregulation of lymphatics, following injury, surgery or chronic inflammation also exacerbates GI disease activity. Understanding the physiological roles played by GI lymphatics is essential to elucidating their underlying contributions to forms of congenital and acquired forms of GI pathology, and will provide novel approaches for therapy. PMID- 20022233 TI - Transmitting the message: intracellular mRNA localization. AB - The intracellular localization of mRNA is an important mechanism for targeting proteins to their sites of function. Localized transcripts play key roles ranging from body axis determination to synapse plasticity, and recent studies highlight how common localized transcripts are in a variety of model organisms. In this review, we consider the full range of known mechanisms for delivering transcripts and anchoring them at their site of function. Finally, we evaluate the progress that has been made in the nervous system in understanding the mechanism and function of mRNA localization and translation regulation. PMID- 20022234 TI - Normalization and harm reduction: research avenues and policy agendas. AB - An affinity between the evidence and arguments for drug normalization and the policy and programme directions favoured by harm reduction is often assumed but seldom critically examined. This commentary looks at parallels and contradictions emerging with respect to different cultures, social settings, types of problems and responses where the match is less than perfect. Mounting evidence of normalization has also led to backlash in some countries and the mobilization of forces reaffirming prohibition. We call for further research on normalization that focuses on substance use, risks, harms, and social context across a broader spectrum of the population, and in a variety of cultures. By emphasizing the most serious harms experienced by persons in the smallest segments of drug using populations, harm reduction often has neglected broader research and policy suggestions that might be implemented to benefit controlled, recreational drug users. Future policy development with respect to normalization will require more research and more serious discussion of its implications for informing the transition toward a global public health approach to substance use. PMID- 20022235 TI - Generating CO(2)-credits through landfill in situ aeration. AB - Landfills are some of the major anthropogenic sources of methane emissions worldwide. The installation and operation of gas extraction systems for many landfills in Europe and the US, often including technical installations for energy recovery, significantly reduced these emissions during the last decades. Residual landfill gas, however, is still continuously produced after the energy recovery became economically unattractive, thus resulting in ongoing methane emissions for many years. By landfill in situ aeration these methane emissions can be widely avoided both, during the aeration process as well as in the subsequent aftercare period. Based on model calculations and online monitoring data the amount of avoided CO(2-eq). can be determined. For an in situ aerated landfill in northern Germany, acting as a case study, 83-95% (depending on the kind and quality of top cover) of the greenhouse gas emission potential could be reduced under strictly controlled conditions. Recently the United Nations Framework Convention on Climate Change (UNFCCC) has approved a new methodology on the "Avoidance of landfill gas emissions by in situ aeration of landfills" (UNFCCC, 2009). Based on this methodology landfill aeration projects might be considered for generation of Certified Emission Reductions (CERs) in the course of CDM projects. This paper contributes towards an evaluation of the potential of landfill aeration for methane emissions reduction. PMID- 20022236 TI - Binary fate decisions in differentiating neurons. AB - Neural cell fate programs must generate an enormous number of neurons with distinct adult functions. The decision to choose one neuronal subtype from two alternatives--a binary fate decision--is one way to diversify neuronal subtypes during nervous system development. Recent progress has been made in describing the genetic programs that define late-stage neuronal identity. Here, we review mechanisms that control how such fate decisions generate two different postmitotic, terminally differentiated neuronal subtypes. We survey examples from Caenorhabditis elegans and Drosophila that demonstrate different modes of binary neuronal fate specification that depend on cell division, lineage, stochastic gene expression, or extracellular signals. Comparison of these strategies reveals that, although organisms use diverse approaches to generate neural diversity, some common themes do exist. PMID- 20022237 TI - Serum 25-hydroxyvitamin D at pregnancy and risk of breast cancer in a prospective study. AB - BACKGROUND: Several laboratory and epidemiological studies have inversely linked endogenous vitamin D and the risk of breast cancer. The acquisition of vitamin D over time on the relative risk (RR) of the disease development is not known. In a longitudinal study, we evaluated the association between vitamin D levels at pregnancy over time with the risk of breast cancer, and pregnancy-associated breast cancer. METHOD: The risk for subsequent development of breast cancer associated with serum 25-hydroxyvitamin (25-OHD) levels was assessed for consecutive (1st and 2nd pregnancy) samples of 100 cases, with mean lag times (micro(t)) of 7.4 and 4.6 years between sampling and the diagnosis, and matched (parity, age, year, season) controls. Pregnancy-associated breast cancer (PABC, 111 case-control pairs, micro(t)=1 year) risk was also studied. Odds ratios (ORs) with 95% confidence intervals (CI) were calculated using the lowest quintile as the reference. RESULTS: Serum 25-OHD level was not associated with an increased risk neither at the 1st nor at the 2nd pregnancy samples (OR=1.4, 95%CI 0.6-3.4; OR 1.4, 95%CI 0.7-2.8, respectively), but was associated with an increased risk of PABC (OR=2.7, 95%CI 1.04-6.7). CONCLUSION: Generally, vitamin D may not be related to breast cancer risk but the increased PABC risk fits the association of vitamin D with the most aggressive cancers, and warrants caution with vitamin D supplementation during pregnancy. PMID- 20022238 TI - Familial risks of breast and prostate cancers: does the definition of the at risk period matter? AB - AIM: 'Being at familial risk' may have different connotations in studies on familial risk of cancer. The register-based definition of a family history considers individuals with an affected relative at familial risk independently of the family member's diagnostic time. Alternatively, the individuals are classified to be at familial risk only after the diagnosis date of their relative, relevant to clinical counselling and screening situations. The aim of this study was to compare familial breast and prostate cancer risks according to the two definitions. PATIENTS AND METHODS: The nationwide Swedish Family-Cancer Database with information on cancers from 1958 to 2006 was used to calculate the hazard ratio of breast and prostate cancers according to family history using Cox regression. Family history was defined considering the number and type of affected relatives and the relative's diagnostic age, respectively. Individuals were considered at familial risk from their entry to the study or, alternatively, from the diagnostic time of the relative. RESULTS: Hazard ratios were equal whether individuals were considered at risk independent of the relative's diagnostic date or only after the relative's diagnostic date. CONCLUSION: These results indicate that studies on familial breast or prostate cancer risk which do not take the relative's diagnosis date into account are applicable to screening and clinical counselling situations. The estimates according to the register based definition are based on larger numbers of patients, which may be crucial for analysis of small groups such as families of multiple cases. PMID- 20022239 TI - Informed decision making on PSA testing for the detection of prostate cancer: an evaluation of a leaflet with risk indicator. AB - BACKGROUND: Population-based screening for prostate cancer (PCa) remains controversial. To help men making informed decisions about prostate specific antigen (PSA) screening a risk indicator (www.uroweb.org) was developed. This risk indicator is embedded in a leaflet that informs men about the pros and cons of PCa screening and enables calculation of the individual risk of having a biopsy detectable PCa. AIM: To assess the effect of providing a leaflet including individualized risk estimation on informed decision making of men, i.e. knowledge about PCa and PSA screening, attitude towards undergoing a PSA test and intention to have a PSA test. METHODS: An intervention study among 2000 men, aged 55-65 years, randomly selected from the population registry of the city of Dordrecht, the Netherlands, in 2008. Men were sent a questionnaire on knowledge of PCa, attitude and intention to have a PSA test. Men without a history of (screening for) PCa were sent the leaflet and Questionnaire 2 within 2 weeks after returning Questionnaire 1. Validated health and anxiety measures were used. RESULTS: One thousand and twenty seven of 2000 men completed Questionnaire 1 (51%), of whom 298 were excluded due to a history of (screening for) PCa. Of the 729 remaining men, 601 completed Questionnaire 2 as well. At the second assessment significantly more men met the requirements of informed decision making (15% versus 33%, p<0.001), more men had relevant knowledge (284/601, 50% versus 420/601, 77%, p<0.001) and the intention to have a PSA test had increased (p<0.001). CONCLUSIONS: Providing information on PCa screening combined with individualized risk estimation enhanced informed decision making and may be used for shared decision making on PSA screening of physicians and patients. PMID- 20022240 TI - Non-steroidal anti-inflammatory drug use in the elderly. AB - The efficacy of non-steroidal anti-inflammatory drugs (NSAIDs) for the treatment of inflammation and pain of various origins is well established. Prescribing these drugs, however, remains a challenge because a great variety of gastrointestinal and cardiovascular safety issues need to be considered, particularly in older patients. Recent recommendations suggest that the prescription of non-selective NSAIDs and/or selective cyclo-oxygenase-2 inhibitors (coxibs) may be appropriate in patients with low gastrointestinal risk (no prior gastrointestinal events, no concomitant treatments with other damaging drugs). Gastroprotection is appropriate in patients with gastrointestinal risk factors and in older patients. In patients at high risk for gastrointestinal and cardiovascular events, however, NSAID or coxib prescriptions are contraindicated. Multidimensional impairment is a crucial point in evaluating the clinical outcome of older patients; thus, a comprehensive geriatric assessment is useful in predicting adverse outcomes, including morbidity and mortality. PMID- 20022241 TI - Separation of furans and carboxylic acids from sugars in dilute acid rice straw hydrolyzates by nanofiltration. AB - This work studied the concentration of hydrolyzates obtained from dilute acid hydrolysis of rice straw using nanofiltration (NF). In order to minimize the Donnan exclusion effect of the membrane, the hydrolyzate solution was controlled at low pH value. Negative retentions of both furans and carboxylic acids were observed. The maximum separation factor of acetic acid over xylose was 49, while the maximum separation factor of acetic acid over arabinose was 52, when the system was operated at pH 2.9 and an applied pressure of 24.5-34.3 bar. The separation factors of inhibitors over glucose became infinity due to the complete retention of glucose. The separation performance decreased when the operating temperature was increased from 25 to 40 degrees C. The flux deterioration was recovered by flushing with 0.01 N of NaOH and water. PMID- 20022242 TI - The solvent influence on the positional selectivity of Novozym 435 during 1,3 diolein synthesis by esterification. AB - The influence of solvents with a wide range of log P (from -0.23 to 4.5) on the positional selectivity of the immobilized lipase Novozym 435 during the esterification of oleic acid with glycerol for 1,3-diolein preparation was investigated. Analysis was performed on the basis of a simplified kinetic model of 1,3-diolein synthesis. The results showed that the preferential selectivity of Novozym 435 to 1-position over 2-position of the glycerol molecular became weaker and weaker with the increasing log P of the solvent. But after one 1-position was acylated, the preferential selectivity to the other 1-position over 2-position would be enhanced strongly for each solvent. The study also revealed that relatively hydrophilic solvent such as t-butanol was an ideal solvent for Novozym 435 catalyzed 1,3-diolein synthesis through esterification of oleic acid with glycerol. PMID- 20022243 TI - Piperazinyl pyrimidine derivatives as potent gamma-secretase modulators. AB - The development of a novel series of piperazinyl pyrimidines as gamma-secretase modulators for potential use in the treatment of Alzheimer's disease is disclosed herein. Optimization of a screening hit provided a series of potent gamma secretase modulators with >180-fold in vitro selectivity over inhibition of Notch cleavage. PMID- 20022245 TI - Shape and dynamics of tip-growing cells. AB - Walled cells have the ability to remodel their shape while sustaining an internal turgor pressure that can reach values up to 10 atmospheres [1-7]. Although it is undisputed that this requires a tight and simultaneous regulation of cell wall assembly and mechanics, previous theoretical studies on tip growth focused either on the mechanical behavior of the cell wall or on its assembly [8-14]. To study the interplay between growth and mechanics in shaping a walled cell, we examine the particularly simple geometry of tip-growing cells [1, 3, 15, 16], which elongate via the assembly and expansion of cell wall in the apical region of the cell. We describe the observed irreversible expansion of the cell wall during growth as the extension of an inhomogeneous viscous fluid shell under the action of turgor pressure, fed by a material source in the neighborhood of the growing tip. This allows us to determine theoretically the radius of the cell and its growth velocity in terms of the turgor pressure and the secretion rate and rheology of the cell wall material. We derive simple scaling laws for the geometry of the cell and find that a single dimensionless parameter, which characterizes the relative roles of cell wall assembly and expansion, is sufficient to explain the observed variability in shapes of tip-growing cells. More generally, our description provides a framework to understand cell growth and remodeling in plants (pollen tubes [17], root hairs, etc. [18]), fungi (hyphal growth [19, 20] and fission and budding yeast [3]), and some bacteria [21], in the context of both tip growth and diffuse growth. PMID- 20022244 TI - Epithelial polarity proteins regulate Drosophila tracheal tube size in parallel to the luminal matrix pathway. AB - Regulation of epithelial tube size is critical for organ function. However, the mechanisms of tube size control remain poorly understood. In the Drosophila trachea, tube dimensions are regulated by a luminal extracellular matrix (ECM). ECM organization requires apical (luminal) secretion of the protein Vermiform (Verm), which depends on the basolateral septate junction (SJ). Here, we show that apical and basolateral epithelial polarity proteins interact to control tracheal tube size independently of the Verm pathway. Mutations in yurt (yrt) and scribble (scrib), which encode SJ-associated polarity proteins, cause an expansion of tracheal tubes but do not disrupt Verm secretion. Reducing activity of the apical polarity protein Crumbs (Crb) suppresses the length defects in yrt but not scrib mutants, suggesting that Yrt acts by negatively regulating Crb. Conversely, Crb overexpression increases tracheal tube dimensions. Reducing crb dosage also rescues tracheal size defects caused by mutations in coracle (cora), which encodes an SJ-associated polarity protein. In addition, crb mutations suppress cora length defects without restoring Verm secretion. Together, these data indicate that Yrt, Cora, Crb, and Scrib operate independently of the Verm pathway. Our data support a model in which Cora and Yrt act through Crb to regulate epithelial tube size. PMID- 20022246 TI - Face adaptation without a face. AB - Prolonged viewing of a stimulus results in a subsequent perceptual bias. This perceptual adaptation and the resulting aftereffect reveal important characteristics regarding how perceptual systems are tuned. These aftereffects occur not only for simple stimulus features but also for high-level stimulus properties. Here we report a novel cross-category adaptation aftereffect demonstrating that prolonged viewing of a human body without a face shifts the perceptual tuning curve for face gender and face identity. This contradicts a central assumption underlying perceptual adaptation: that adaptation depends on physical similarity between how the adapting and the adapted features are perceived. Additionally, this aftereffect was not due to response bias, because its dependence on adaptation duration resembled traditional perceptual aftereffects. These body-to-face adaptation results demonstrate that bodies alone can alter the tuning properties of neurons that code for the gender and identity of faces. More generally, these results reveal that high-level perceptual adaptation can occur when the property or features being adapted are automatically inferred rather than perceived in the adapting stimulus. PMID- 20022247 TI - Regulating alternative lifestyles in entomopathogenic bacteria. AB - Bacteria belonging to the genera Photorhabdus and Xenorhabdus participate in a trilateral symbiosis in which they enable their nematode hosts to parasitize insect larvae. The bacteria switch from persisting peacefully in a nematode's digestive tract to a lifestyle in which pathways to produce insecticidal toxins, degrading enzymes to digest the insect for consumption, and antibiotics to ward off bacterial and fungal competitors are activated. This study addresses three questions: (1) What molecular signal triggers antibiotic production in the bacteria? (2) What small molecules are regulated by the signal? And (3), how do the bacteria recognize the signal? Differential metabolomic profiling in Photorhabdus luminescens TT01 and Xenorhabdus nematophila revealed that L-proline in the insect's hemolymph initiates a metabolic shift. Small molecules known to be crucial for virulence and antibiosis in addition to previously unknown metabolites are dramatically upregulated by L-proline, linking the recognition of host environment to bacterial metabolic regulation. To identify the L-proline induced signaling pathway, we deleted the proline transporters putP and proU in P. luminescens TT01. Studies of these strains support a model in which acquisition of L-proline both regulates the metabolic shift and maintains the bacterial proton motive force that ultimately regulates the downstream bacterial pathways affecting virulence and antibiotic production. PMID- 20022249 TI - Computer simulations from a finite-element model for wound contraction and closure. AB - Some implications from a simplified finite-element model are given in this study. The model takes into account the sequential steps of wound contraction, angiogenesis and wound closure. An innovation in the present study is the combination of these partially overlapping processes, yielding novel insights into the process of wound healing, such as geometry related influences, and could be used to investigate the influence of local injection of hormones that stimulate partial processes occurring during wound healing. These insights can be used to improve wound-healing treatments. The model consists of nonlinearly coupled diffusion-reaction and visco-elastic equations, in which transport, production and decay of oxygen, growth factors and various cell types. The present paper provides results of the healing of deep wounds under several regimes of endothelial and epithelial cell migration, and the results are interpreted in a biological sense. PMID- 20022248 TI - A broadly conserved pathway generates 3'UTR-directed primary piRNAs. AB - BACKGROUND: Piwi-interacting RNAs (piRNAs) are approximately 24-30 nucleotide regulatory RNAs that are abundant in animal gonads and early embryos. The best characterized piRNAs mediate a conserved pathway that restricts transposable elements, and these frequently engage a "ping-pong" amplification loop. Certain stages of mammalian testis also accumulate abundant piRNAs of unknown function, which derive from noncoding RNAs that are depleted in transposable element content and do not engage in ping-pong. RESULTS: We report that the 3' untranslated regions (3'UTRs) of an extensive set of messenger RNAs (mRNAs) are processed into piRNAs in Drosophila ovaries, murine testes, and Xenopus eggs. Analysis of different mutants and Piwi-class immunoprecipitates indicates that their biogenesis depends on primary piRNA components, but not most ping-pong components. Several observations suggest that mRNAs are actively selected for piRNA production for regulatory purposes. First, genic piRNAs do not accumulate in proportion to the level of their host transcripts, and many highly expressed transcripts lack piRNAs. Second, piRNA-producing mRNAs in Drosophila and mouse are enriched for specific gene ontology categories distinct from those of simply abundant transcripts. Third, the protein output of traffic jam, whose 3'UTR generates abundant piRNAs, is increased in piwi mutant follicle clones. CONCLUSIONS: We reveal a conserved primary piRNA pathway that selects and metabolizes the 3'UTRs of a broad set of cellular transcripts, probably for regulatory purposes. These findings strongly increase the breadth of Argonaute mediated small RNA systems in metazoans. PMID- 20022250 TI - The effects of soybean agglutinin binding on the corneal endothelium and the re establishment of an intact monolayer following injury--a short review. AB - This short review summarizes the localization and effects of the plant lectin soybean agglutinin (SBA) on the injured and non-injured organ-cultured rat corneal endothelium. Although the tissue exists as a non-cycling monolayer on the posterior corneal surface a circular freeze injury promotes wound repair as cells initiate DNA synthesis, mitosis and migration. As a result, by 24 h post-injury, endothelial cells express a surface protein that binds SBA in a diffuse punctate pattern, which by 48 h after injury, becomes confined to the cell periphery. As healing proceeds, SBA binding dramatically declines, such that, only scattered binding is observed by 72 h after wounding. In non-injured organ-cultured endothelia, weak SBA binding appears 24 h after explanation but becomes prominently detected around the cell periphery by 48 h. Incubating injured or non injured endothelia in SBA leads to alterations in their cellular appearance due to the fact that lectin exposure results in the disruption of the actin cytoskeleton. Although this does not affect migration, treatment with either SBA or N-acetylgalactosamine (the SBA binding sugar) does interfere with the reestablishment of cell-cell contact. It is postulated that the surface protein that binds SBA is expressed during conditions that are stressful to the tissue. During organ-culture the protein's appearance suggests a cellular response to explantation in order to enhance or maintain monolayer integrity. In wound repair its appearance may serve to establish preliminary cell-cell contact during the restoration of the endothelial monolayer. PMID- 20022251 TI - Changes in hip joint muscle-tendon lengths with mode of locomotion. AB - We have reported that peak hip extension is nearly identical in walking and running, suggesting that anatomical constraints, such as flexor muscle tightness may limit the range of hip extension. To obtain a more mechanistic insight into mobility at the hip and pelvis we examined the lengths of the muscle-tendons units crossing the hip joint. Data defining the three-dimensional kinematics of 26 healthy runners at self-selected walking and running speeds were obtained. These data were used to scale and drive musculoskeletal models using OpenSIM. Muscle-tendon unit (MTU) lengths were calculated for the trailing limb illiacus, rectus femoris, gluteus maximus, and biceps femoris long head and the advancing limb biceps femoris and gluteus maximus. The magnitude and timing of MTU length peaks were each compared between walking and running. The peak length of the right (trailing limb) illiacus MTU, a pure hip flexor, was nearly identical between walking and running, while the maximum length of the rectus femoris MTU, a hip flexor and knee extensor, increased during running. The maximum length of the left (leading limb) biceps femoris was also unchanged between walking and running. Further, the timing of peak illiacus MTU length and peak contralateral biceps femoris MTU length occurred essentially simultaneously during running, at a time during gait when the hamstrings are most vulnerable to stretch injury. This latter finding suggests exploring the role for hip flexor stretching in combination with hamstring stretching to treat and/or prevent running related hamstring injury. PMID- 20022252 TI - Custom-made lateral femoral condyle replacement for traumatic bone loss: a case report. AB - We report the case of a 24 year-old patient who underwent a novel treatment for a lateral femoral condyle fracture. The fracture was associated with extensive joint line depression and not considered suitable for conventional fixation techniques. Existing reconstructive options for such situations include unicondylar osteoarticular allograft, arthrodesis and arthroplasty. However, these techniques all present significant disadvantages, particularly in the management of active patients. We report our medium-term results following reconstruction using a custom-made lateral femoral condyle hemiarthroplasty replacement. Follow-up at 48 months revealed an excellent, pain-free level of function, with an Oxford Knee Score of 46/48, a Knee Society knee score of 87/100 and a functional score of 100/100. Radiographs demonstrated no evidence of prosthesis loosening or migration and no erosion of the lateral tibial plateau. The technique allows preservation of the remaining normal joint surface of the femur that may promote earlier and better restoration of function. Furthermore, the isolated condyle hemi-replacement maximises bone preservation, facilitating future anticipated revisions. The procedure presents an attractive alternative to other surgical options and their attendant problems. Further investigation into this technique is required before widespread adoption, though such studies will be compromised by the relative rarity of patients in whom the technique is indicated. PMID- 20022253 TI - Isolation, structure, and bioactivities of abiesadines A-Y, 25 new diterpenes from Abies georgei Orr. AB - Twenty-five new (abiesadines A-Y, 1-25) and 29 known (26-54) diterpenes were isolated from the aerial parts of Abies georgei. Abiesadine A (1) is a novel 8,14 seco-abietane, while abiesadine B (2) is a novel 9,10-seco-abietane. The structures of the new compounds were established on the basis of spectroscopic data analysis. Manool (52) showed the strongest effect against LPS-induced NO production in RAW264.7 macrophages with the IC(50) value of 11.0microg/mL. In another anti-inflammatory assay against TNFalpha-triggered NF-kappaB activity, (12R,13R)-8,12-epoxy-14-labden-13-ol (54) exhibited the strongest effect (IC(50)=8.7microg/mL). For antitumor assays, pomiferin A (26) and 8,11,13 abietatriene-7alpha,18-diol (29) both showed the most significant activity against LOVO cells (IC(50)=9.2microg/mL). While 7-oxocallitrisic acid (46) exhibited significant cytotoxicity against QGY-7703 tumor cells (IC(50)=10.2microg/mL). PMID- 20022254 TI - Development of a neutron-activated concrete powder reference material. AB - In this paper, the development of a neutron-activated concrete powder reference material is described. The material originated from core samples taken from a concrete bioshield of a decommissioned nuclear reactor which ceased operation almost 30 years ago after approximately 20 years of operation. The assigned values, which were in the Bq g(-1) range, for the radionuclides in the material were determined by a 'consensus' method from measurements made by 33 organisations from 15 countries. The measurements were made within a wider test exercise (the NPL Environmental Radioactivity Proficiency Test Exercise 2008). Assigned specific activity values were obtained for (60)Co, (133)Ba, (152)Eu and (154)Eu and indicative values were obtained for (3)H (total), (3)H (leachable), (14)C, (40)K, (55)Fe and (63)Ni. PMID- 20022255 TI - In-situ gamma spectrometry in environmental monitoring. AB - In-situ gamma spectrometry (scintillation or semiconductor) can be used effectively for monitoring natural and man-made radionuclide concentrations, together with the corresponding photon fields, in the environment and in workplaces. It is applied in operational and emergency monitoring of nuclear facilities, waste storage facilities and the uranium industry, in radioactive contamination measurements and mapping, environmental, radiohygienic and radiation safety studies, etc. Methods for processing and interpreting data, experimental techniques (ground or airborne arrangement), calibration and verification and examples of applications are discussed in this paper. PMID- 20022256 TI - Improved method for the calculation of the counting efficiency of electron capture nuclides in liquid scintillation samples. AB - The methods to compute the counting efficiency of electron-capture nuclides in liquid scintillation counting have been improved in several previous studies. The main improvements comprise a more realistic treatment of the ejection of photoelectrons and subsequent rearrangement processes in the atomic shell as well as a more detailed atomic rearrangement model. The latter was realized in the MICELLE code by means of a new stochastic approach. This new model was also developed to account for energy deposits within micelles. The recent improvements have now been combined in an updated version of the MICELLE code, which also makes the computation of the counting efficiency of complex decay schemes possible. In this paper, we describe and discuss recent extensions and improvements of the models and further corrections. The calculated counting efficiencies of selected radionuclides are compared with the experimental data obtained by liquid scintillation counting. For the measurements, we use standard solutions, which were calibrated by other methods. PMID- 20022257 TI - Emission probabilities of gamma-rays from 238Np and their use for determination of the thermal neutron capture cross section of 237Np. AB - The relevant absolute gamma-ray emission probabilities from the beta-decay of (238)Np were measured by means of alpha- and gamma-spectroscopic techniques. We obtained values of (25.6+/-0.4)%, (8.9+/-0.2)% and (18.8+/-0.3)% for the 984.45-, 1025.87- and 1028.54-keV gamma-rays, respectively, in agreement with the previous measured ones. These intensities were used to deduce the thermal neutron capture cross section of (237)Np for which a value of (182.2+/-4.5)b is obtained higher by 11% than the recommended value. PMID- 20022258 TI - Standardisation of positron-emitters in gas with the NPL primary gas counting system. AB - A primary standard for positron-emitters in gas has been developed. The method involves internal gas proportional counting and the use of the PENELOPE Monte Carlo code to determine corrections for counting losses. The development work was carried out using (11)C, although the method can be applied to other positron emitters. The results were compared with measurements of (11)C (in solution) carried out using a secondary standard re-entrant ionisation chamber previously calibrated with reference to absolute counting techniques. PMID- 20022259 TI - The 222Rn standard system established at IFIN-HH, Romania. AB - The paper presents the (222)Rn Standard System realized at the Radionuclide Metrology Laboratory from IFIN-HH, Romania. It contains a Pylon solid (226)Ra source, and a glass circuit for circulation and recovery of (222)Rn in glass ampoules, at the 77K temperature. The radon can be recovered both in ampoules with liquid scintillator (LS), for absolute standardization of (222)Rn by liquid scintillation counting (LSC) (Cassette et al., 2006), and in evacuated ampoules as gas, simultaneously or by the transfer of radon from gas into LS ampoules, in order to establish the traceability chain. The absolute standardization consists in the registration of the double coincidence counting rate in LS, due to the whole sequence of short life components of the (222)Rn chain, in equilibrium conditions. The main correction applied to the measurement results is due to the decay of (214)Po during the extendable dead time of the system. The following procedure was applied to take it into account. The value of the base duration of the dead time was precisely measured and used for the data corrections. The measurements of each source were repeated during a period of several days, and the decay curve parameters were calculated. If the dead time value and the correction formulae, presented throughout the paper, were adequate, then the re determination of the (222)Rn half life, T(1/2)=(3.8232+/-0.0008) d, using the decay curve of the corrected counting rates should be correct. On the other hand, the effective dead time values obtained by parallel calculations were compared. The paper presents the results obtained in the standardization of several sources, by LSC, and the evaluation of the uncertainties associated to the method. PMID- 20022261 TI - Arterial pressure variation and goal-directed fluid therapy. PMID- 20022260 TI - Sequential enrichment of sulfated glycans by strong anion-exchange chromatography prior to mass spectrometric measurements. AB - Structural characterization of sulfated glycans through mass spectrometry (MS) has been often limited by their low abundance in biological materials and inefficient ionization in the positive-ion mode. Here, we describe a microscale method for sequentially enriching sulfated glycans according to their degree of sulfation. This method is based on modifying the binding ability of strong anion exchange material through the use of different sodium acetate concentrations, thus enabling fairly selective binding and a subsequent elution of different glycans according to their degree of sulfation. Before this enrichment, the negative charge on the sialic acid, which is commonly associated with such glycans, was eliminated through permethylation that is used to enhance the positive-ion mode matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-MS) signal for all glycans. This enrichment approach minimizes competitive ionization between sulfated and neutral glycans, as well as that between sulfated species with a different degree of sulfation. The described method was initially optimized using sulfated oligosaccharide standards, while its potential has been verified for the sulfated N-glycans originated from the bovine thyroid-stimulating hormone (bTSH), a glycoprotein possessing mono- and disulfated N-glycans. This enhancement of the MALDI-MS signal facilitates analysis of some otherwise undetected components. PMID- 20022262 TI - Milrinone improves diastolic function in coronary artery bypass surgery as assessed by acoustic quantification and peak filling rate: a prospective randomized study. AB - OBJECTIVE: To compare the effects of a bolus dose of milrinone, 50 microg/kg, to placebo on diastolic function (active relaxation) in patients undergoing on-pump coronary artery bypass grafting (CABG). DESIGN: Prospective, randomized, double blind, placebo-controlled study. SETTING: University hospital. PARTICIPANTS: Twenty-four patients with stable angina and left ventricular ejection fraction >30%, scheduled for elective CABG using cardiopulmonary bypass (CPB), were included. INTERVENTION: Patients were randomized to receive either 50 microg/kg of milrinone (n = 12) or placebo (n = 12) after aortic declamping. MEASUREMENTS AND MAIN RESULTS: The diastolic function of the left ventricle (LV) was measured as peak filling rate (dA/dt [maximal diastolic area change over time]) with transesophageal echocardiography (TEE) using acoustic quantification (AQ) before CPB and 10 minutes after termination of CPB. The normalized peak filling rate (dA/dt)/EDA was also calculated. Active relaxation was statistically significantly increased in the milrinone group compared with the placebo group after CPB. CONCLUSION: Patients undergoing CABG surgery and treated with milrinone after aortic declamping had better diastolic function following cardiopulmonary bypass. PMID- 20022263 TI - Activation of JAK-STAT and nitric oxide signaling as a mechanism for donor heart dysfunction. AB - BACKGROUND: Donor heart dysfunction (DHD) precluding procurement for transplantation occurs in up to 25% of brain-dead (BD) donors. The molecular mechanisms of DHD remain unclear. We investigated the potential role of myocardial interleukin (IL)-6 signaling through the JAK2-STAT3 pathway, which can lead to the generation of nitric oxide (NO) and decreased cardiac myocyte contractility. METHODS: Hearts were procured using standard technique with University of Wisconsin (UW) solution from 14 donors with a left ventricular (LV) ejection fraction of <35% (DHD). Ten hearts with normal function (NF) after BD served as controls. LV IL-6 was quantitated by enzyme-linked immunoassay (ELISA) and JAK2-STAT3 signaling was assessed by expression of phosphorylated STAT3. Inducible NO synthase (iNOS) and caspase-3 were measured by activity assays. RESULTS: Myocardial IL-6 expression was 8-fold greater in the DHD group vs NF controls. Phosphorylated STAT3 expression was 5-fold higher in DHD than in NF, indicating increased JAK2-STAT3 signaling. LV activity of iNOS was 2.5-fold greater in DHD than in NF. LV expression of the pro-apoptotic gene Bnip3 and caspase-3 activity were 3-fold greater in the DHD group than in the NF group. CONCLUSIONS: Myocardial IL-6 expression is significantly higher in the setting of DHD compared with hearts procured with normal function. This may lead to increased JAK2-STAT3 signaling and upregulation of iNOS, which has been shown to decrease cardiac myocyte contractility. Increased NO production may also lead to increased apoptosis through upregulation of Bnip3 gene expression. Increased iNOS signaling may be an important mechanism of DHD and represents a novel therapeutic target to improve cardiac function after BD. PMID- 20022264 TI - Exercise improvement and plasma biomarker changes with intravenous treprostinil therapy for pulmonary arterial hypertension: a placebo-controlled trial. AB - BACKGROUND: Pulmonary arterial hypertension (PAH) remains a poorly understood and frequently lethal disease with few treatment options. METHODS: We conducted a placebo-controlled trial of intravenous treprostinil, a prostacyclin analog, in treatment-naive PAH patients. During 12 weeks of treatment with treprostinil or placebo, we quantified 6-minute walk distance (6MW), clinical symptoms and 11 cytokines/growth factors. RESULTS: Forty-two of 44 study patients had idiopathic/familial PAH in New York Heart Association (NYHA) Class III. Treprostinil increased 6MW by a placebo-corrected median of 83 meters (p = 0.008; mean increase 93 +/- 42 meters), reduced Borg score by a median 2.0 units (p = 0.02), and improved NYHA class by a median of 1.0 (p = 0.02). There was a trend toward improved survival with treprostinil (p = 0.051). Baseline plasma angiopoietin-2 (Ang-2), vascular endothelial growth factor (VEGF), matrix metalloproteinase-9 (MMP-9) and platelet-derived growth factor (PDGF) were elevated compared with reported normal ranges. Treatment with treprostinil was associated with decreased Ang-2 levels. Improvement in 6MW distance after treatment was associated with reductions in Ang-2 and MMP-9 levels. Most of the cytokines and growth factors studied were not abnormal with disease nor did they change with treatment. CONCLUSIONS: We conclude that treprostinil treatment significantly improved exercise capacity, dyspnea and functional class. Several plasma proteins that might track disease were abnormal at baseline, and changes were associated with improved exercise capacity. PMID- 20022266 TI - Concreteness of thinking and self-focus. AB - The present study used the experience sampling method to detect fluctuations in thinking, such as self-focus or concreteness in daily life, and to examine their relationship with depressive symptoms and concurrent negative affect. Thirty-one undergraduates recorded their negative affect, ruminative self-focus, and concreteness of thinking eight times a day for 1 week. Multilevel modeling showed that individuals with increasing levels of depression showed lower levels of concreteness in their daily thinking. Further analysis revealed a significant positive association between momentary ruminative self-focus and concurrent negative affect only with low concreteness of thinking. These results suggested that individuals with increasing levels of depression chronically process self related information on an abstract level, which reflects a malfunction of their self-regulatory cycle and might serve to maintain or even exacerbate dysphoric moods. PMID- 20022265 TI - Increased erythrocyte C4D is associated with known alloantibody and autoantibody markers of antibody-mediated rejection in human lung transplant recipients. AB - BACKGROUND: Immune responses to mismatched donor human leukocyte antigens (HLA) are important in the pathogenesis of chronic rejection. This study evaluated whether erythrocyte-bound C4d (E-C4d) is associated with known alloimmune and autoimmune markers of antibody-mediated rejection after human lung transplantation (LTx). METHODS: Flow cytometry was used to analyze 22 LTx recipients and 15 healthy individuals for E-C4d. Development of antibodies to donor-mismatched HLA (donor-specific antibody [DSA]) and antibodies to HLA were determined using the solid-phase method by Luminex. Development of antibodies to self-antigens, K-alpha-1-tubulin (KA1T) and collagen V (Col-V), were measured by enzyme-linked immunosorbent assay. C3d deposition in lung biopsy specimens was determined by immunohistochemical staining. RESULTS: Percent E-C4d (%E-C4d) levels were 19.9% in LTx patients vs 3.7% in healthy individuals (p = 0.02). DSA+ patients had higher E-C4d levels than DSA- patients (34.1% vs 16.7%, p = 0.02). In 5 patients with preformed anti-HLA, E-C4d levels were not significantly different vs 13 patients without detectable anti-HLA (p = 0.1). E-C4d levels were higher in patients who developed antibodies to KA1T (p = 0.02) and Col-V (p = 0.03). Recipients with C3d-positive tissue deposition had higher E-C4d levels than patients with C3d-negative biopsy results (p = 0.01). CONCLUSIONS: Increased %E-C4d levels are found in patients with positive DSA, high antibody titers to KA1T and Col-V, and have C3d+ lung biopsy findings. Therefore, %E-C4d can serve as a potential marker for antibody-mediated rejection after LTx. PMID- 20022267 TI - Osteopontin overexpression predicts poor prognosis of upper urinary tract urothelial carcinoma. AB - OBJECTIVES: Studies indicate overexpression of osteopontin (OPN) promotes carcinogenesis, progression and metastasis of multiple human malignancies. However, the function of OPN in urothelial carcinoma (UC) of the upper urinary tract has not been investigated. This study evaluates the clinical significance of OPN expression in upper urinary tract UC. MATERIALS AND METHODS: One hundred and ten cases (median age = 64, range = 24-84 years) of renal pelvic or ureter UC were retrospectively reviewed in this study. OPN expression were evaluated by immunohistochemistry staining on paraffin-embedded section of the tumor and scored by two qualified pathologists. RESULTS: High OPN expression was found in 54 (49.1%) of the cancer specimens. OPN expression was not significantly correlated with tumor T stage (P = 0.761), N stage (P = 0.339) or grade (P = 0.349). However, OPN expression was differently expressed by gender (P = 0.012) and cancer location (P = 0.026). OPN expression did not correlate with bladder recurrence-free (P = 0.661) or extra-bladder recurrence-free (P = 0.787) survival, but high OPN expression was a significant predictor for cancer-specific survival (P = 0.014). CONCLUSION: Our findings indicated that higher OPN expression is a potential biomarker to predict patient survival. Further study is necessary to investigate the role of OPN in the carcinogenesis of upper urinary tract UC. PMID- 20022268 TI - Neoadjuvant docetaxel/estramustine prior to radical prostatectomy or external beam radiotherapy in high risk localized prostate cancer: a phase II trial. AB - BACKGROUND: Patients with locally advanced or organ confined, high risk, prostate cancer are at significant risk of having disease recurrence despite definitive local therapy. We evaluated the 2-year progression-free survival of subjects treated with chemotherapy administered prior to definitive therapy with surgery or radiation. PATIENTS AND METHODS: Patients (n = 24) with locally advanced and high risk localized prostate cancer were treated with neoadjuvant docetaxel 36 mg/m2 i.v. weekly for 3 weeks and estramustine 140 mg orally 3 times daily for 3 consecutive days every 28 days prior to definitive treatment with prostatectomy or radiation. RESULTS: All evaluable patients, except 1, completed the proposed cycles of neoadjuvant chemotherapy with minimal dose reductions or delays. Of the 22 evaluable patients, 12 underwent radical prostatectomy and 10 underwent external beam radiation therapy. Twenty-one of 22 patients achieved a prostate specific antigen (PSA) reduction > 25%. There were no pathologic complete responses. With a median follow-up of 24 months, the 2-year progression-free survival was 45%. CONCLUSIONS: Our findings support the safety, tolerability, and efficacy of neoadjuvant chemotherapy in patients with men with high risk, locally advanced prostate adenocarcinoma, although the relative contributions of androgen deprivation therapy and docetaxel cannot be determined. The effectiveness of neoadjuvant chemotherapy in preventing prostate cancer relapses should be studied in a randomized trial. PMID- 20022269 TI - Detection of inguinal lymph node involvement in penile squamous cell carcinoma by 18F-fluorodeoxyglucose PET/CT: a prospective single-center study. AB - BACKGROUND: The extent of lymph node involvement is the most relevant prognostic factor in patients with penile cancer. OBJECTIVE: To prospectively analyze the diagnostic accuracy of 18F-FDG-PET/CT-scan in the assessment of inguinal lymph node involvement in patients with invasive penile carcinoma. PATIENTS AND METHODS: Thirty-five patients with invasive penile carcinoma were staged prospectively by 18F-FDG-PET/CT-scan, and blindly evaluated by 2 nuclear medicine physicians. In total, lymph node involvement was assessed in 70 inguinal groins. Reference standard was either histology or clinical follow-up with a minimum of 31 months (mean: 48.4 months; range: 31-68 months). RESULTS: 18-FDG-PET/CT showed a sensitivity of 88.2% and a specificity of 98.1%. Positive predictive value (PPV) was 93.8%, while negative predictive value (NPV) was 96.3%. In two groins, metastasis of 5 and 7 mm were missed by PET/CT scan. CONCLUSION: 18F-FDG-PET/CT is a promising staging tool in assessing the inguinal lymph node involvement of patients with penile carcinoma. Integration of PET/CT scanning into preoperative staging algorithms may avoid surgical staging in selected patients. PMID- 20022270 TI - Interactions of hemoglobin Lepore (deltabeta hybrid hemoglobin) with various hemoglobinopathies: A molecular and hematological characteristics and differential diagnosis. AB - Hemoglobin (Hb) Lepore is a variant consisting of two alpha-globin and two deltabeta-globin chains. In heterozygote, it is associated with clinical findings of thalassemia minor but interactions with other hemoglobinopathies can lead to various clinical phenotypes. Using a combination of Hb-HPLC, Hb-capillary electrophoresis and DNA analyses, we have identified 14 patients with Hb Lepore Hollandia including eight heterozygotes, two double heterozygotes with alpha(+) thalassemia, two compound heterozygotes with Hb E (initially diagnosed as Hb E beta-thalassemia) and two previously undescribed conditions of double heterozygote for Hb Lepore/Hb Constant Spring and Hb Lepore/alpha(0)-thalassemia, both associated with higher levels of Hb F and lower levels of Hb Lepore. Hematological and molecular features of these patients are presented along with those observed in four other Thai individuals encountered with heterozygous Hb Lepore-Washington-Boston. Haplotype analysis of the beta-globin gene cluster showed that all Hb Lepore-Hollandia genes were associated with a single haplotype not described previously in other populations, (- + - + + - +) whereas the four Hb Lepore-Washington-Boston genes were associated with haplotypes (+ - - - - + /+) (N=1) and (+ - - - - - +) (N=3), data indicating multiple origins of these two variants. Hb Lepore may not be uncommon in the Thai and other Asian populations and both hematological and molecular studies are required for accurate diagnosis. To facilitate rapid epidemiological, diagnostic screening and differentiation of the two Hb Lepore defects, a simple assay based on multiplex PCR has been developed. PMID- 20022271 TI - Guidelines for the restart of imiglucerase in patients with Gaucher disease: recommendations from the European Working Group on Gaucher disease. PMID- 20022272 TI - Group epitope mapping considering relaxation of the ligand (GEM-CRL): including longitudinal relaxation rates in the analysis of saturation transfer difference (STD) experiments. AB - In the application of saturation transfer difference (STD) experiments to the study of protein-ligand interactions, the relaxation of the ligand is one of the major influences on the experimentally observed STD factors, making interpretation of these difficult when attempting to define a group epitope map (GEM). In this paper, we describe a simplification of the relaxation matrix that may be applied under specified experimental conditions, which results in a simplified equation reflecting the directly transferred magnetisation rate from the protein onto the ligand, defined as the summation over the whole protein of the protein-ligand cross-relaxation multiplied by with the fractional saturation of the protein protons. In this, the relaxation of the ligand is accounted for implicitly by inclusion of the experimentally determined longitudinal relaxation rates. The conditions under which this "group epitope mapping considering relaxation of the ligand" (GEM-CRL) can be applied were tested on a theoretical model system, which demonstrated only minor deviations from that predicted by the full relaxation matrix calculations (CORCEMA-ST) [7]. Furthermore, CORCEMA-ST calculations of two protein-saccharide complexes (Jacalin and TreR) with known crystal structures were performed and compared with experimental GEM-CRL data. It could be shown that the GEM-CRL methodology is superior to the classical group epitope mapping approach currently used for defining ligand-protein proximities. GEM-CRL is also useful for the interpretation of CORCEMA-ST results, because the transferred magnetisation rate provides an additional parameter for the comparison between measured and calculated values. The independence of this parameter from the above mentioned factors can thereby enhance the value of CORCEMA-ST calculations. PMID- 20022273 TI - Bloch simulations with intra-voxel spin dephasing. AB - A common problem in simulations of MRI-experiments based on the numerical solution of the Bloch equations is the finite number of isochromats used in the calculations. This usually results in false or spurious signals and is a source of various differences between calculated and experimentally obtained data. In this paper, we are proposing a technique representing each sample voxel by a central and three additional isochromats, slightly shifted in orthogonal directions from center, thus providing a linear approximation of intra-voxel dephasing. This approach allows for further improvement and precision of the calculated NMR signal and virtually avoids the problem related to an finite set of isochromats. Here we provide details of the algorithm together with examples of simulations which prove the efficiency of this approach. PMID- 20022274 TI - Gastric smooth muscle hamartoma in a cat. AB - An 11-year-old cat presented for evaluation of intermittent vomiting, constipation and hyporexia of 3 weeks duration. Ultrasonographic and endoscopic examination revealed a soft tissue mass adjacent to the lower gastro-esophageal sphincter. Surgical excision of the mass was successfully performed resulting in a resolution of clinical signs. Histologically the mass was consistent with a smooth muscle hamartoma. At follow-up 7 months after surgery, the cat remained free from clinical signs. PMID- 20022275 TI - Calculating and controlling the error of discrete representations of Pareto surfaces in convex multi-criteria optimization. AB - A discrete set of points and their convex combinations can serve as a sparse representation of the Pareto surface in multiple objective convex optimization. We develop a method to evaluate the quality of such a representation, and show by example that in multiple objective radiotherapy planning, the number of Pareto optimal solutions needed to represent Pareto surfaces of up to five dimensions grows at most linearly with the number of objectives. The method described is also applicable to the representation of convex sets. PMID- 20022276 TI - [A response by A. Agostini to the article of R. Lousquy et al.: State of the art about teaching hysteroscopy to gynaecologist surgeons in France and in Europe. CONFORM investigation into training surgeons on the hysteroscopic placement of microinserts]. PMID- 20022277 TI - [Microsurgical tubal desterilization: an efficient procedure. Evaluation of a 42 cases cohort according to Guzick's biparametric method]. AB - OBJECTIVE: Tubal ligation tends to be a contraceptive method used by younger women. Regrets may then become more frequent. Although less tough today, microsurgical tubal reversal is a reliable technique and we report results taking into account exposure time and curing rate. PATIENTS AND METHODS: We studied 42 cases operated from 1990 and 1997. The patients were selected following the same protocol and operated according to the same surgical way. The results were analyzed by Guzick biparametric method, and comparison assessed by Log-rank test. We compared results depending on age, type of sterilization (mechanical and non mechanical), duration of laparotomic time, and interval between sterilization and reversal. RESULTS: There is an overall hope of 80% of pregnancy at 49 months. Even if there is no statistical difference between the different parameters studied, there is a trend to better reversal results after mechanical sterilization and intervention duration less than 90 minutes. DISCUSSION AND CONCLUSION: Microsurgical tubal reversal is a trustworthy technique, which may be offered to patients regretting their sterilization. Analyzing results by Guzick method is interesting, but insufficiently used although more precise to express a curing (= pregnancy) result than a simple Kaplan-Meier surviving curve. Microsurgical reversal may be proposed to carefully selected patients, even after 40 years. PMID- 20022278 TI - [Late post-partum eclampsia: a case report]. AB - Post-partum eclampsia is an unfrequent entity (11 to 44 % of eclampsia), which can sometimes occur late, beyond the 48th hour. We report a case of late post partum eclampsia which occurred 12 days after an uncomplicated pregnancy and delivery. Post-partum eclampsia differs from antepartum by a pauci symptomatic or unexpected context. The mortality rate is comparable with that of the ante-partum eclampsia. Brain MRI appears to be the investigation of choice in case of uncertain diagnosis. Magnesium sulfate treatment should be considered under the same conditions as during pregnancy. Identifying and informing high-risk women would allow early diagnosis, which may improve the prognosis of this atypical type of eclampsia. PMID- 20022279 TI - [PubMed for the hopeless!]. PMID- 20022280 TI - [Endometriosis of the ischio-rectal excavation at the contact of the sciatic nerve: a case report of neurolysis by pararectal incision]. AB - Localisation of endometriosis on the sciatic nerve is exceptional. We report the case of a patient presenting an endometriotic nodule of the left ischio-rectal excavation, with an extension contiguous to the sciatic nerve, responsible of invalidating sciatalgia. Two laparoscopies did not allow to localise the lesion. Finally the endometriotic nodule was treated by a direct access of the left ischio-rectal excavation through a pararectal incision. In this article we discuss the means to localise such lesion and the surgical approach to propose. PMID- 20022281 TI - [Oocyte donation as in France]. AB - Oocyte donation, initially proposed in agonadal women, saw indications expand to ovarian deficiencies and failures of in vitro fertilization (IVF), resulting in a significant increasing demand. The recruitment of oocyte donors is a critical issue for all countries that have allowed this practice. The French legislation, with the laws of bioethics, is clearly the most restrictive of European countries, imposing an absolute free gift from mother. The different solutions in the neighboring countries are analysed and in particular the interpretations made in respect of gratuity and compensation. Motivating donors (spontaneous, relational, or by reciprocity), but also motivating the medical teams can organize a program of oocyte donation in France. The authors present their results of three years experience, demonstrating that this system is possible in the current legislative framework. PMID- 20022282 TI - [Efficiency of antagonist IVF cycle programming by estrogens]. AB - OBJECTIVE: Assess the efficiency of estradiol programming in In Vitro Fertilization (IVF) with antagonists by comparing with classical long luteal agonist protocol. PATIENTS AND METHODS: It is a prospective randomized study, comparing 426 cycles in the arm estradiol antagonist with 412 cycles in the arm long agonist. Estradiol 4 mg/day begins on the 25th day of the previous cycle and continues during the menses until the first day of the stimulation which is from Thursday to Sunday whatever the beginning of the menses. The luteal protocol use Decapeptyl 0,1mg which begins on the 20th day of the previous cycle. RESULTS: Our two populations are similar. No pick-up has been done on Sunday. We have got significantly less oocytes and embryos in estradiol-antagonist (6,8+/-5,3 vs 7,6+/-5,7) and (3,7+/-3,2 vs 4,1+/-3,6) respectively. The ongoing pregnancy rate is comparable in the two groups: 28,6 % for estradiol antagonist 27,9 % for agonist for the whole population and 37 % vs 34,8 % respectively when at least one top embryo was transferred. DISCUSSION AND CONCLUSION: Programming antagonist cycles with estradiol allows the organization of the center; it is easy to implement and seems to give results as good as a long agonist protocol. PMID- 20022283 TI - Sonochemical synthesis of nano-scale mixed-ligands lead(II) coordination polymers as precursors for preparation of PbO and PbBr(OH) nano-structures; thermal, structural and X-ray powder diffraction studies. AB - Nano-structures of two new lead(II) two-dimensional coordination polymers, [Pb(2)(2-pyc)(2)(I)(2)](n) (1) and [Pb(2)(2-pyc)(2)(Br)(2)](n) (2), {2-Hpyc=2 pyridinecarboxilic acid } were synthesized by a sonochemical method that produce the coordination polymers at nano-size. The new nano-structures were characterized by scanning electron microscopy, X-ray powder diffraction, IR spectroscopy and elemental analyses. Compounds 1 and 2 were structurally characterized by single crystal X-ray diffraction and consist of two-dimensional polymeric units. The thermal stability of compounds 1 and 2 were studied by thermal gravimetric and differential thermal analyses. PbO and PbBr(OH) nano structures were obtained by calcination of the nano-structures of compounds 1 and 2 at 400 degrees C, respectively. PMID- 20022284 TI - Ultrasound-assisted three-component synthesis of 3-(5-amino-1H-pyrazol-4-yl)-3-(2 hydroxy-4,4-dimethyl-6-oxocyclohex-1-enyl)indolin-2-ones in water. AB - Indium(III) chloride was found to be an efficient catalyst for the synthesis of 3 (5-amino-1H-pyrazol-4-yl)-3-(2-hydroxy-4,4-dimethyl-6-oxocyclohex-1-enyl)indolin 2-ones by one-pot, three-component reaction of dimedone, 1H-pyrazol-5-amines and isatins in water under ultrasonic irradiation. The advantages of this method are the use of a readily available catalyst, easy workup, excellent yields, and the use of water as a solvent that is considered to be relatively environmentally benign. PMID- 20022285 TI - Health status and health behaviours in neighbourhoods: a comparison of Glasgow, Scotland and Hamilton, Canada. AB - Health status has been demonstrated to vary by neighbourhood socioeconomic status (SES). However, neighbourhood effects may vary between countries. In this study, neighbourhood variations in health outcomes are compared across four socially contrasting neighbourhoods in Glasgow, Scotland and Hamilton, Ontario Canada. Data came from the 2001 wave of the West of Scotland Twenty-07 Longitudinal Study and a 2000/2001 cross-sectional survey conducted in Hamilton. The results of the comparison point to important variations in the relationship between neighbourhood SES and health. While both cities display a socioeconomic gradient with respect to various measures of health and health behaviours, for some outcome measures the high SES neighbourhoods in Glasgow display distributions similar to those found in the low SES neighbourhoods in Hamilton. Our results suggest that a low SES neighbourhood in one country may not mean the same for health as a low SES neighbourhood in another country. As such, country context may explain the distribution of health status and health behaviours among socially contrasting neighbourhoods, and neighbourhood variations in health may be context specific. PMID- 20022286 TI - Individual and socio-environmental determinants of overweight and obesity in Urban Canada. AB - Overweight/obesity represent a significant public health problem in Canada and abroad. The objective of this paper is to identify potential associations between overweight/obesity and individual as well as socio-environmental determinants. The data sources used are the 2003 Canadian Community Health Survey and the Desktop Mapping Technologies Incorporated database. Geographical Information Systems are first employed to create neighbourhood-level variables such as neighbourhood walkability and fast food accessibility. Multivariate analysis is then applied to estimate the relative effects of individual- and neighbourhood level risk-factors of overweight/obesity. Results demonstrate the important role of the built-environment after adjustment for demographic, socio-economic and behavioural characteristics. Findings support the rationale that reversing current trends will require a multifaceted public health approach where interventions are developed from the individual- to the neighbourhood-level, with a particular focus on altering obesogenic environments. PMID- 20022287 TI - Polymer synthesis by enzymatic catalysis. AB - Using biocatalysts as enzymes for in vitro polymer synthesis is a relatively new (third) stream that has become popular in the past two decades, following a first stream using catalysts of acids, bases, radical species, and so on, used since 1920s, and a second stream using catalysts of transition metals and rare metals since 1950s. Enzymatic catalysis is environmentally benign, yet allows the synthesis of various natural and unnatural polymers that have well-defined structures with controlled stereochemistry, regioselectivity, and chemoselectivity. This review will focus on recent developments in enzyme catalyzed polymer synthesis in the areas of polysaccharides, polyesters, and polyaromatics, as well as polymer modification. PMID- 20022288 TI - Molecular imaging. PMID- 20022289 TI - Adapting genomics to study the evolution and ecology of agricultural systems. AB - In the face of global change, agriculture increasingly requires germplasm with high yields on marginal lands. Identifying pathways that are adaptive under marginal conditions is increasingly possible with advances at the intersection of evolutionary ecology, population genetics, and functional genomics. Trait-based (reverse ecology) approaches have connected flowering time in Arabidopsis thaliana to single alleles with environment-specific effects. Similarly, genetic dissection of rice flooding tolerance enabled the production of near-isogenic lines exhibiting tolerance and high yields. An alternative gene-forward (forward ecology) approach identified candidate genes for local adaptation of Arabidopsis lyrata to heavy-metal rich soils. A global perspective on plant adaptation and trait correlations provides a foundation for breeding tolerant crops and suggests populations adapted to marginal habitats be conservation priorities. PMID- 20022290 TI - The humanitarian impact of plant biotechnology: recent breakthroughs vs bottlenecks for adoption. AB - The deployment of genetically engineered (GE) crops in developing countries is regarded by some as a sinister manifestation of 'big business' in science. What is often overlooked, and sometimes even deliberately ignored by opponents of the technology, is that many researchers working in the field are not motivated by profits but by a desire to see such crops applied to humanitarian purposes. GE crops could help to address many of the world's most challenging, interrelated problems, including hunger, malnutrition, disease, and poverty. However, this potential will not be realized if the major barriers to adoption - which are political rather than technical - are not overcome. PMID- 20022291 TI - Bi-exponential decay of Eu(III) complexed by Suwannee River humic substances: spectroscopic evidence of two different excited species. AB - The bi-exponential luminescence decay of europium (III) complexed by Suwannee River fulvic acid (SRFA) and humic acid (SRHA), is studied in time-resolved luminescence spectroscopy using two different gratings at varying delay after the laser pulse, increasing accumulation time in order to obtain comparable signals. The two hypotheses found in the literature to interpret this bi-exponential decay are (i) a back transfer from the metal to the triplet state of the organic ligand and (ii) the radiative decay of two different excited species. It is shown that evolutions of the (5)D(0)-->(7)F(0) and (5)D(0)-->(7)F(2) luminescent transitions are occurring between 10 and 300 micros delay. First, the (5)D(0)-->(7)F(0) transition is decreasing relative to the (5)D(0)-->(7)F(1) showing a slightly greater symmetry of the 'slow' component, and is also slightly red shifted. Second, a slight modification of the (5)D(0)-->(7)F(2) transition is also evidencing a slightly different ligand field splitting. No significant modification of the (5)D(0)-->(7)F(1) magnetic dipole, which is less susceptible to symmetry changes, is noted in line with expectations. The (5)D(0)-->(7)F(0) transitions are adjusted with either one or two components. The use of a simple component fit seems to be well adapted for representing an average comportment of these heterogeneous compounds, and a two-component fit constrained by the bi exponential decay parameters and accumulation times yields in the proposition of the spectra for the fast and slow components. PMID- 20022292 TI - Solvatochromic behaviours and structure-spectra relationships of 4-carboxyl-2,6 dinitrophenylazohydroxynaphthalenes. AB - Comprehensive electronic absorption spectra of a new dye series, 4-carboxyl-2,6 dinitrophenylazohydroxynaphthalenes have been investigated in solvents of varying polarities. The solvent dependent UV-vis spectral shifts were analysed using some solvent physical parameters such as refractive index, dielectric function, hydrogen bonding acceptor ability, orientation of polarization and others. The observed spectral shifts were correlated with different solute-solvent interaction mechanisms using simple and multiple linear regression analyses. The results of the curve fitting coefficients enabled us to classify the various interactions of solvents with the dyes and relate the solvatochromic behaviours to the substituent effects on the dye molecules. Charge-transfer complexation occurring between one of the congeners and N,N'-dimethylformamide was extensively studied and discovered to be both concentration- and temperature-dependent. The electronic character and the chemical nature of the solvents as well as the chemical nature of the other substituents, apart from the common hydroxyl group, are important factors for the observed solvatochromic properties of the 4 carboxyl-2, 6-dinitrophenylazohydroxynaphthalenes. PMID- 20022293 TI - A study of the conformational stability and the vibrational spectra of 2,3 dichloro-1-propanol. AB - The conformational stability and the three rotor internal rotations in 2,3 dichloro-1-propanol were investigated at DFT-B3LYP/6-311+G**, MP2/6-311+G** and MP4(SDQ) levels of theory. From the calculated potential energy surface, ten distinct minima were located all of which were predicted to have real frequencies at the B3LYP level of theory. The calculated lowest energy minima in the potential curves of the molecule were predicted to correspond to the Ggg and Gtg1 structures. The observed broad and very intense infrared band centered at about 3370 cm(-1) supports the existence of the strong intermolecular H-bonding in 2,3 dichloro-1-propanol. The equilibrium constants for the conformational interconversion in the molecule were estimated from the calculated Gibb's energies at the B3LYP/6-311+G** level of calculation and found to correspond to an equilibrium mixture of about 49% Ggg, 27% Gtg1, 5% Ggt and 5% Tgg conformations at 298.15K. PMID- 20022294 TI - Study on the fluorescence characteristics of carbon dots. AB - Herein, we prepared water-soluble fluorescent carbon dots with diameter about 1.5 nm from cheap commercial lampblack. These fluorescent carbon nanoparticles are stable toward photobleaching and stable in water for more than half a year without fluorescence decrease. In order to improve its fluorescence properties, we passivated these nanoparticles with bisamino-terminated polyethylene glycol (PEG(1500 N)). Therefore, both fluorescence quantum yield and lifetime increased after this progress. In addition, the passivated carbon dots were more inert to solvent than the bare one and showed different responses to pH change. PMID- 20022295 TI - Correlation between bocavirus infection and humoral response, and co-infection with other respiratory viruses in children with acute respiratory infection. AB - BACKGROUND: Human bocavirus (HBoV), a recently discovered virus, is prevalent among children with respiratory tract infection throughout the world. Co infection was frequently found in HBoV-positive patients. Thus, whether HBoV is responsible for the respiratory disease is still arguable. OBJECTIVES: A comprehensive study was carried out to integrate clinical and virological prevalence in HBoV-positive outpatient children, and to determine genetic and serologic characteristics of HBoV in Shanghai, China. STUDY DESIGN: Nasal/throat swabs and sera were obtained over a 2-year period from 817 children with respiratory tract infection to examine the presence of HBoV and its co-infection. The seroepidemiology of HBoV was studied by ELISA and Western blot against the capsid protein VP2-based fragment. Persistence of HBoV was also analyzed in 12 pairs of return-visit cases. RESULTS: HBoV was identified in 96 samples (11.8%). The co-infection rate with other respiratory viruses was 51%. IgM was detected in 55.7% of HBoV RT-PCR-positive patients, and in 72.7% of those who had high viral genome load. In addition, persistent viral DNA positivity was detected in 10 of 12 HBoV-positive cases tested, an average of 14 days later, and one child was still HBoV-positive after 31 days. CONCLUSION: HBoV was found frequently in children with respiratory tract symptoms associated with other respiratory viruses, and persisted in the respiratory tract and in serum and urine. The presence of IgM was significantly more prevalent in viremic patients and those diagnosed with high load of HBoV DNA in nasal/throat swabs. PMID- 20022296 TI - Performance of laboratory diagnostics for the detection of influenza A(H1N1)v virus as correlated with the time after symptom onset and viral load. AB - BACKGROUND: To diagnose influenza A(H1N1)v virus infection, accurate and rapid detection are important. However, there is scanty data on the performance of various laboratory diagnostics. OBJECTIVE: To compare the performance of rapid antigen test (RAT), viral culture and RT-PCR for the detection of influenza A(H1N1)v virus and to correlate their performance with the time after symptom onset and viral load. STUDY DESIGN: From May 1, 2009 to June 25, 2009, respiratory samples were collected from 5740 individuals suspected of having influenza A(H1N1)v infection. The performance of viral culture and RT-PCR were investigated and correlated with the time after symptom onset. The sensitivity of RAT ESPLINE influenza A & B-N (Fujirebio Inc, Tokyo) was evaluated using a subset of 60 samples from patients diagnosed as having influenza A(H1N1)v infection. RESULTS: Using respiratory samples from 587 patients diagnosed with influenza A(H1N1)v infection, comparison of laboratory diagnostics showed viral culture and RT-PCR gave comparable results with overall sensitivity of 93.9% and 98.1%, respectively. For RAT, when testing a subset of 60 samples collected < or =3 days following symptom onset, the sensitivity was 62%. CONCLUSIONS: Although viral shedding is prolonged and of higher titre in influenza A(H1N1)v infection, RAT showed a low sensitivity of 62% among patients presenting < or =3 days after symptom onset. Viral culture showed comparable performance with RT-PCR and with sensitivity better than that documented for seasonal influenza. PMID- 20022297 TI - Variability and recombination of clinical human cytomegalovirus strains from transplantation recipients. AB - BACKGROUND: Human cytomegalovirus (HCMV) is the first cause of viral infection in immunocompromised transplanted patients. OBJECTIVES: Here, five HCMV genes were studied to investigate the existence of recombination events in clinical strains ex vivo. STUDY DESIGN: Sequencing and phylogenetic analysis were conducted on 21 strains from 16 renal and 5 lung transplant recipients. RESULTS: Nucleotidic polymorphism ranged from 6.6% (US3) to 12% (UL40), with a significant proportion of missense mutations (39-69%), some of which could have a functional impact. Analysis of the concatenated sequence (4804 nucleotides for each strain) evidenced two clusters of sequences presenting a reticulate topology suggestive of recombination events (SplitsTree). Phi-test pointed numerous phylogenetically conflicting signals indicating a high statistical probability of recombination. The subsequent bootscan analysis was consistent with these data. CONCLUSIONS: These results reinforce the prominent role of recombination in HCMV evolutionary history and adaptation to its host. PMID- 20022298 TI - Sleep-disordered breathing symptoms frequency and growth among prematurely born infants. AB - BACKGROUND: Children who were born prematurely are at higher risk for sleep disordered breathing (SDB) compared to their same-age peers who were born fullterm. OBJECTIVE: The objective of the present study was to assess the frequency of SDB symptoms and effects on growth among preterm infants while they are still infants, with a goal of identifying risk factors to facilitate prevention and early intervention. METHODS: The Louisville pediatric SDB risk survey was administered to the primary caretakers of prematurely born infants attending the Neonatal Follow-Up Clinic at West Virginia University Children's Hospital. RESULTS: Participation was 100% among 173 consecutive patients invited to participate in the study. At 9.13 months corrected age, 8.1% of infants born at a mean of 31.6 weeks gestation were reported to snore > or = 3 days/week, a rate consistent with diagnosis of sleep-disordered breathing among older children. A composite of nine parent-reported symptoms was created based on factor analysis. Birth weight and size for gestational age at birth did not differ between snoring groups or correlate with the composite score. But a significant negative correlation between the composite risk for SDB score and current weight for adjusted age percentile score indicate that infants with higher SDB symptom profiles have lower weight for age (r=-.18, p=.028). CONCLUSIONS: SDB symptoms are detectable among infants born preterm, while they are still infants. Because of their preferential risk for SDB and its somatic consequences, a primary research goal should be description of the natural history of SDB and identification of modifiable risk factors and treatment options. PMID- 20022299 TI - REM sleep behavior disorder in 703 sleep-disorder patients: the importance of eliciting a comprehensive sleep history. AB - OBJECTIVES: The aim of our study was to evaluate the frequency of REM sleep behavior disorder (RBD) in a mixed sleep laboratory population and to assess potential associations. Moreover, we investigated referral diagnoses of patients subsequently diagnosed with RBD and assessed the frequency of incidental RBD. METHODS: Charts and polysomnographic reports of 703 consecutive patients comprising the full spectrum of ICSD-2 sleep disorders [501 males, 202 females; mean age, 51.0+/-14.1 years (range: 10-82 years)] were carefully reviewed. The vast majority of patients were adults (98.7%). Patients were categorized into those with and without RBD. For associations, all concomitant sleep and neurological diagnoses and medications were evaluated. RESULTS: Thirty-four patients (4.8%) were diagnosed with RBD (27 men; 7 women, mean age, 57.7+/-12.3 years). RBD was idiopathic in 11 patients (1.6%; 9 men) and symptomatic in 23 patients (3.3%; 18 men) secondary to Parkinsonian syndromes (n=11), use of antidepressants (n=7), narcolepsy with cataplexy (n=4), and pontine infarction (n=1). Six out of 34 patients were referred for suspected RBD, 20 reported RBD symptoms only on specific questioning, and 8 patients had no history of RBD but showed typical RBD behavioral manifestations in the video-polysomnography. Logistic regression analysis revealed significant associations between RBD and the presence of Parkinsonian syndromes (odds ratio [OR] 16.8, 95%CI: 6.4-44.1; P<0.001), narcolepsy with cataplexy (OR 10.7, 95%CI: 2.9-40.2; P<0.001), SSRI use (OR 3.9, 95%CI: 1.6-9.8; P=0.003), and age (OR 1.5/10-year increase, 95%CI: 1.0 2.0; P=0.039). CONCLUSION: In this population of 703 consecutive sleep-disorder patients, RBD was uncommon. Its etiology was predominantly symptomatic. The majority of RBD patients reported RBD symptoms on specific questioning only, underlining the importance of eliciting a comprehensive sleep history for the diagnosis of RBD. PMID- 20022300 TI - Ultrastructure and 18S rDNA phylogeny of Apoikia lindahlii comb. nov. (Chrysophyceae) and its epibiontic protists, Filos agilis gen. et sp. nov. (Bicosoecida) and Nanos amicus gen. et sp. nov. (Bicosoecida). AB - Three heterotrophic stramenopiles--Apoikia lindahlii comb. nov. (Chrysophyceae), Filos agilis gen. et sp. nov. (Bicosoecida), and Nanos amicus gen. et sp. nov. (Bicosoecida)--were isolated from acidic peat bogs. The biflagellate A. lindahlii forms loose irregular colonies from which swimming cells may detach, and produces extensive mucilaginous material containing bacterial cells. Phylogenetic analyses of small subunit rDNA sequences demonstrated that A. lindahlii branches within the Chrysophyceae. While A. lindahlii is an obligate heterotroph, ultrastructural observations revealed a leukoplast in the perinuclear region. The pico-sized uniflagellates F. agilis and N. amicus were isolated from separate lakes and within the mucilage of A. lindahlii, suggesting their close associations in natural habitats. In SSU rDNA phylogenies, F. agilis and N. amicus were closely related to the bicosoecids Adriamonas, Siluania, Paramonas, and Nerada. While Filos, Nanos, and Siluania are similar in light microscopic features, their SSU rDNA gene sequences differed significantly (>8% differences) and were not monophyletic. Both F. agilis and N. amicus have a cytostome/cytopharynx particle ingestion apparatus. Bacterial cells and material similar to the mucilage of A. lindahlii occurred within the food vacuole of F. agilis and N. amicus. The nature of association between A. lindahlii and its epibiontic bicosoecids is discussed. PMID- 20022301 TI - Influence of posture and body type on the experience of exercise-related transient abdominal pain. AB - The aim of this study was to evaluate the influence of posture and body type on the experience of exercise-related transient abdominal pain (ETAP). Postural and somatotype assessments were performed on 104 active males and 55 active females aged 18.6+/-0.4 years (mean+/-SD) and were correlated against their self-reported experience of ETAP. Individuals demonstrating kyphosis were more likely to be susceptible to ETAP (p<0.01). Among the individuals susceptible to ETAP, the extent of kyphosis and lordosis influenced the pain severity (p<0.05). There was no relationship between any measure of body type and ETAP. The findings indicate that postural abnormality, particularly in the thoracic region, influences the experience of ETAP. PMID- 20022302 TI - Influence of sex on the "Athlete's Heart" in trained cyclists. AB - Compared to females, male endurance athletes have generally been considered to demonstrate greater values of cardiac mass and volume (the "athlete's heart"). However, studies addressing this issue have frequently failed to match training volumes between groups or provided inadequate adjustment of variables for body size and composition. This study compared echocardiographic anatomic features in 8 female and 8 male competitive cyclists with a similar training history. Conforming to most previous reports, left ventricular mass and end diastolic dimension, adjusted for fat free body mass and body surface area, respectively, were greater in the males (3.56+/-0.83gkg(-1) versus 2.50+/-0.38gkg(-1); 41.7+/ 2.2mm and 37.4+/-2.5mm per BSA(0.5)). This study indicated that when training volume as well as body size and composition are considered, male endurance athletes exhibit greater cardiac dimensions and mass compared to their female counterparts. PMID- 20022303 TI - Application and student evaluation of a Clinical Progression Portfolio: a pilot. AB - Clinical practicums are often limited by a lack of meaningful communication between nursing students and registered nurses (RNs). This pilot study evaluated the utility of the Clinical Progression Portfolio (CPP) to enable students to learn how to initiate engagement with their RNs and to develop their capacity as students to learn. The study employed a descriptive survey design, with a convenience sample of second-year Bachelor of Nursing (BN) students in Brisbane, Australia. Questionnaires were completed by 129 students from 20 clinical practicum groups. Students who used the CPP were more favourable in their usefulness ratings (-rpb=0.531, p<0.001) and, furthermore, those that used the CPP most frequently were also more favourable (r=0.555, p<0.001). Students thought the CPP helped clarify learning and target appropriate practicum opportunities. When used, the CPP was an important part of practicum, used frequently and considered useful. The CPP format met the needs of students as it was pocket-sized. Overall, students reported that the CPP was a useful learning and communication tool as it provided them direction in how they might maximise opportunities to address their learning needs. PMID- 20022304 TI - Coping, sense of coherence and the dimensions of affect in patients with chronic heart failure. AB - BACKGROUND: Living with chronic heart failure has an impact on several important dimensions of an individual's life. A patient's use of coping strategies may influence his or her health condition and emotional well-being. AIM: To investigate factors that may relate to the coping strategies used by individuals with chronic heart failure and how the coping strategies are associated with positive and negative affect. METHODS: A cross-sectional research design was used. The participants provided demographic data and filled out three questionnaires: Sense of Coherence scale, Brief COPE and Positive Affect Negative Affect Schedule. RESULTS: No differences in relation to coping strategies were found with regard to New York Heart Association class. Substance use was associated with gender and age. Sense of coherence was negatively associated with denial, behavioural disengagement, venting and self-blame, and positively associated with acceptance. It was found that avoidant coping positively and sense of coherence negatively, predicted negative affect. Problem focused coping positively predicted positive affect. Socially supported coping predicted both negative and positive affect. CONCLUSION: The present study found that the employment of different coping strategies and sense of coherence had an impact on affect and therefore also the emotional well-being among patients with chronic heart failure. PMID- 20022305 TI - Rituximab treatment overcomes reduction of regulatory iNKT cells in patients with rheumatoid arthritis. AB - Invariant natural killer T (iNKT) cells are a subset of T cells that recognize glycolipid antigens presented by the CD1d molecule. Accumulating evidences showed that iNKT cells are implicated in the regulatory mechanisms that control autoimmunity. We evaluated the number of circulating iNKT cells in patients with rheumatoid arthritis (RA) by flow cytometry and performed a longitudinal analysis of iNKT cell frequency in RA patients who were given an anti-CD20 therapy. Significantly lower iNKT cell numbers were measured in the blood from RA patients compared to healthy individuals (p<0.0001) and low iNKT cell frequencies were rather associated with an active disease. In RA patients who received rituximab treatment, iNKT cell number was increased in relation to the clinical outcome. We demonstrated that the number of iNKT cells is altered in RA patients and that following rituximab therapy, clinical remission of RA is associated with an increase of iNKT cell frequency. PMID- 20022306 TI - Defining the DNA mismatch repair-dependent apoptotic pathway in primary cells: evidence for p53-independence and involvement of centrosomal caspase 2. AB - Many studies have shown that DNA mismatch repair (MMR) has a role beyond that of repair in response to several types of DNA damage, including ultraviolet radiation (UV). We have demonstrated previously that the MMR-dependent component of UVB-induced apoptosis is integral to the suppression of UVB-induced tumorigenesis. Here we demonstrate that Msh6-dependent UVB-induced apoptotic pathway is both activated via the mitochondria and p53-independent. In addition, we have shown for the first time that caspase 2, an initiator caspase, localizes to the centrosomes in mitotic primary mouse embryonic fibroblasts, irrespective of MMR status and UVB treatment. PMID- 20022307 TI - Analysis of 8-oxo-7,8-dihydro-2'-deoxyguanosine by ultra high pressure liquid chromatography-heat assisted electrospray ionization-tandem mass spectrometry. AB - Increased amounts of reactive oxygen species (ROS), generally termed oxidative stress, are frequently hypothesized to be causally associated with many diseases. Analyses of 8-oxo-7,8-dihydro-2'-deoxyguanosine (8-oxo-dG) in DNA and urine are widely used biomarkers for oxidative stress. Over the years it became clear that analysis of 8-oxo-dG in DNA is challenging due to artifactual formation during sample work up. The present study demonstrates that 8-oxo-dG can be measured reliably and accurately when appropriate precautions are taken. First, the presence of an antioxidant, metal chelator, or free radical trapping agent during sample preparation improves reproducibility. Second, sample enrichment by HPLC fraction collection was used to optimize sensitivity. Third, heat assisted electrospray ionization (HESI) eliminated potential interferences and improved assay performance and sensitivity. Subsequently, the UPLC-HESI-MS/MS method was applied to show the biphasic dose response of 8-oxo-dG in H(2)O(2)-treated HeLa cells. Application of this method to human lymphocyte DNA (n=156) gave a mean+/ SD endogenous amount of 1.57+/-0.88 adducts per 10(6) dG, a value that is in agreement with the suggested amount previously estimated by European Standard Committee on Oxidative DNA Damage (ESCODD) and others. These results suggest that the present method is well suited for application to molecular toxicology and epidemiology studies investigating the role of oxidative stress. PMID- 20022308 TI - Ultra-performance liquid chromatography-tandem mass spectrometry analysis of the bioactive components and their metabolites of Shaofu Zhuyu decoction active extract in rat plasma. AB - A rapid, sensitive and selective ultra-performance liquid chromatography electrospray ionization tandem mass spectrometric (UPLC-ESI-MS/MS) method was developed for analysis and identification of the bioactive components and their metabolites in rat plasma following oral administration Shaofu Zhuyu decoction active extract. The analysis was carried out on an AcQuity UPLC chromatographic instrument and a QTOF mass spectrometer using positive and negative electrospray ionization (ESI), respectively. The results showed that sixteen peaks were detected and twelve peaks, including flavones, organic acids and terpene glycosides, were identified by comparing with reference compounds. Furthermore, nine metabolites, including quercetin glucuronide sulfates, quercetin diglucuronides, isorhamnetin sulfates, isorhamnetin glucosides, and isorhamnetin glucuronides were detected and identified in rat plasma based on the mass fragmentation behaviors and literature reports. These results provided a meaningful basis for evaluating the bioactive components and their action mechanisms of complex traditional Chinese medicines (TCMs). PMID- 20022309 TI - Determination of free and liposomal amphotericin B in human plasma by liquid chromatography-mass spectroscopy with solid phase extraction and protein precipitation techniques. AB - Amphotericin B is available in various drug delivery systems such as cholesteryl sulfate complex, as lipid complex, and as liposomal formulation. The separation and measurement of free drug (drug which is not bound with liposomal lipids) and liposomal drug (drug which is entrapped in liposomes) in the human plasma after injection of liposomal Amphotericin B is of prime importance due to toxicity concerns. A robust, specific and sensitive method has been developed to effectively separate and then quantify the free drug and liposomal drug, present in human plasma. This method utilizes solid phase extraction Oasis HLB cartridges, which retains the free drug and the liposomal Amphotericin B was eluted from the cartridge in first step. The eluted liposomal Amphotericin B was then extracted from lipids by protein precipitation method using 2% dimethylsulfoxide (DMSO) in acetonitrile. After separation and extraction, the quantification of free and liposomal fractions of Amphotericin B was performed by HPLC-MS-MS technique. The chromatographic separation was performed using Chromolith Performance RP 18e column. The mobile phase composed of 5 mM ammonium acetate, methanol and acetonitrile and a gradient elution program was used. The calibration curves were found to be linear for free Amphotericin B (0.25-15.0 microg/ml) and liposomal Amphotericin B (1.0-100.0 microg/ml). The recovery was about 96% for free Amphotericin B and about 92% for liposomal Amphotericin B. Recoveries were consistent over the linearity ranges defined. The intra-batch and inter-batch accuracy and precision fulfilled the international requirements. The stability of free and liposomal Amphotericin B was assessed under different storage conditions. PMID- 20022310 TI - Clinical experience with exenatide in a routine secondary care diabetes clinic. AB - Exenatide use in type 2 diabetes is limited in routine clinical practice. We examined a cross-section of 90 patients. Mean weight and HBA(1c) were 114.9+/ 20.6 kg, 10.3+/-2.1% at initiation; 108.0+/-15.3 kg (p<0.0001), 9.0+/-2.1% (p<0.001) at 3 months; 109.2+/-18.2 kg (p<0.0001), 9.5+/-2.3% (p=0.08) at 6 months. Exenatide appears effective in reducing HBA(1c) and weight. PMID- 20022311 TI - On the division of working memory and long-term memory and their relation to intelligence: A latent variable approach. AB - The present study examined the extent to which working (WM) and long-term memory (LTM) reflect the same, related, or completely different constructs and how they relate to other cognitive ability constructs. Participants performed various WM, recall, recognition, general fluid (gF) and general crystallized intelligence (gC) measures. Confirmatory factor analyses suggested that the memory measures could be grouped into three separate yet correlated factors (WM, recall, and recognition) and that these factors were strongly related to gF, but were related less so with gC. Furthermore, it was found that the common variance from the three memory factors could be accounted for by a higher-order memory factor which was strongly related to gF, but less so with gC. Finally, structural equation modeling suggested that both the variance common to the WM tasks and the variance common to all the memory tasks accounted for a unique variance in gF. These results are interpreted within an embedded process model of memory and suggest that WM and LTM tasks measure both shared and unique processes, which are important for intelligence. PMID- 20022313 TI - Lymph node dissection in the surgical management of atypical endometrial hyperplasia. AB - OBJECTIVE: The aim of this study was to evaluate the effect of lymph node dissection in patients with atypical endometrial hyperplasia. STUDY DESIGN: Patients undergoing surgical management of atypical endometrial hyperplasia during the study period were retrospectively identified. Clinical and pathologic information was analyzed. RESULTS: Eighty-eight patients comprised our cohort. Median age was 54 years (range, 37-85 years). Sixty-seven patients had lymph node dissection at the time of surgery for atypical endometrial hyperplasia, whereas 21 did not. Twenty-five of 88 (28.4%) had endometrial carcinoma on final uterine pathology. Stages were as follows: 4 IA, 15 IB, 3 IC, 2 IIB, and 1 IIIC. Surgical outcomes were not statistically significant between staged and unstaged groups. Information from lymph node dissection influenced management decisions in 7 of the 25 (28%) cancer patients. CONCLUSION: Lymph node dissection did not adversely affect surgical outcomes in patients with atypical endometrial hyperplasia. Because many of these patients have concurrent endometrial cancer, we recommend consideration of lymph node dissection in atypical endometrial hyperplasia patients undergoing definitive surgical treatment. PMID- 20022312 TI - Effect of antenatal betamethasone on maternal and fetal amino acid concentration. AB - OBJECTIVE: To determine the concentration of amino acids in women receiving the first course of antenatal betamethasone and to evaluate the umbilical venous and arterial amino acid concentrations at the time of elective cesarean section after betamethasone administration. STUDY DESIGN: Blood samples were collected from 34 pregnant women at risk of premature delivery before and 24 and 48 hours after the first course of betamethasone. In addition, maternal and cord blood samples were collected in 13 women undergoing an elective cesarean section between 24 and 192 hours after betamethasone. RESULTS: Maternal amino acid concentrations were significantly increased after the first dose of betamethasone. Overall total amino nitrogen increased 17.5% 24 hours after betamethasone administration and 20.5% after 48 hours. The concentration of most amino acids was increased both in the umbilical vein and artery after maternal betamethasone administration. CONCLUSION: The concentration of maternal and fetal amino acids increases significantly after betamethasone administration. PMID- 20022314 TI - Does intraoperative spillage of benign ovarian mucinous cystadenoma increase its recurrence rate? AB - OBJECTIVE: To report a higher than estimated recurrence rate of benign mucinous cystadenomas after complete resection of the first one, and to assess potential risk factors for recurrence after complete surgical excision. STUDY DESIGN: We retrospectively reviewed all cases of women who underwent either laparoscopic or laparotomic removal of benign mucinous adnexal cysts by either adnexectomy or cystectomy in our institution between 1996 and 2006. RESULTS: Included were the data of 42 women who fulfilled study entry criteria. Three of them (7.1%) underwent a second operation because of a recurrence of the lesion. A significantly higher rate of women who had cyst recurrence had undergone cystectomy as opposed to adnexectomy (P < .05). Intraoperative rupture of cysts during cystectomy was also significantly associated with cyst recurrence (P < .03). CONCLUSION: Mucinous cystadenoma recurrence is apparently not as rare as reported in the literature. Intraoperative cyst rupture and cystectomy instead of adnexectomy emerged as being two risk factors for recurrence. PMID- 20022317 TI - [Thick caliber errors in the application of statistical tests]. PMID- 20022315 TI - Dose-response effects of betamethasone on maturation of the fetal sheep lung. AB - OBJECTIVE: Glucocorticoid administration to women in preterm labor improves neonatal mortality and morbidity. Fetal exposure to glucocorticoid levels higher than those appropriate to the current gestational stage has multiple organ system effects. Some, eg, fetal hypertension, are maximal at lower than the clinical dose. We hypothesized that the clinical dose has supramaximal lung maturational effects. STUDY DESIGN: We evaluated the full, half, and quarter clinical betamethasone dose (12 mg/70 kg or 170 microg/kg intramuscularly twice 24 hours apart) on fetal sheep lung pressure volume curves (PVC) after 48 hours' exposure at 0.75 gestation. We measured key messenger RNAs and protein products that affect lung function and total lung dipalmitoyl phosphatidyl choline. RESULTS: Full and half doses had similar PVC and total lung dipalmitoyl phosphatidyl choline effects. Messenger RNA for surfactant proteins A, B, and D and elastin increased in a dose-dependent fashion. CONCLUSION: Half the clinical betamethasone dose produces maximal PVC improvement in fetal sheep at 0.75 gestation. PMID- 20022318 TI - Continuous time wavelet entropy of auditory evoked potentials. AB - In this paper, the continuous time wavelet entropy (CTWE) of auditory evoked potentials (AEP) has been characterized by evaluating the relative wavelet energies (RWE) in specified EEG frequency bands. Thus, the rapid variations of CTWE due to the auditory stimulation could be detected in post-stimulus time interval. This approach removes the probability of missing the information hidden in short time intervals. The discrete time and continuous time wavelet based wavelet entropy variations were compared on non-target and target AEP data. It was observed that CTWE can also be an alternative method to analyze entropy as a function of time. PMID- 20022319 TI - Real-time seizure prediction from local field potentials using an adaptive Wiener algorithm. AB - Approximately 30% of individuals with epilepsy have refractory seizures that cannot be controlled by current pharmacological treatment measures. For such patients, responsive neurostimulation prior to a seizure may lead to greater efficacy when compared with current treatments. In this paper, we present a real time adaptive Wiener prediction algorithm implemented on a digital signal processor to be used with local field potential (LFP) recordings. The hardware implementation of the algorithm enables it to be a miniaturized portable system that could be used in a hand-held device. The adaptive nature of the algorithm allows the seizure data to be compared with baseline data occurring in the recent past rather than a preset value. This enhances the sensitivity of the algorithm by accounting for the time-varying dynamics of baseline, inter-ictal and ictal activity. The Wiener algorithm was compared to two statistical-based naive prediction algorithms. ROC curves, area over ROC curves, predictive power, and time under false positives are computed to characterize the algorithm. Testing of the algorithm via offline Matlab analysis on kainate-treated rats results in prediction of seizures about 27 s before clinical onset, with 94% sensitivity and a false positive rate of 0.009 min(-1). When implemented on a real-time TI C6713 signal processor, the algorithm predicts seizures about 6.7s before their clinical onset, with 92% sensitivity and a false positive rate of 0.08 min(-1). These results compare favorably with those obtained in similar studies in terms of sensitivity and false positive rate. PMID- 20022320 TI - Pentraxin 3 in amniotic fluid as a marker of intra-amniotic inflammation in women with preterm premature rupture of membranes. AB - OBJECTIVE: To determine whether amniotic fluid levels of pentraxin 3 (PTX3) are of value in the prenatal diagnosis of acute histological chorioamnionitis in preterm premature rupture of membranes (PPROM). METHODS: Forty pregnant women with PPROM between 24 and 36 weeks of pregnancy without (n=21) and with (n=19) histological chorioamnionitis (PPROM group) and 42 women between 16 and 20 weeks of pregnancy (midtrimester group) were included in the study. We compared amniotic fluid PTX3 levels in the PPROM group with versus without histological chorioamnionitis, and between the PPROM and the midtrimester groups using nonparametric tests (Mann-Whitney test), given the non-normal distribution of the analyte. RESULTS: Patients with histological chorioamnionitis had a significantly higher median amniotic fluid PTX3 concentration than patients without the histological signs of chorioamnionitis (3.69ng/mL [0.51-106.8] versus 0.8ng/mL [0.36-121.0]; P=0.015). Patients in the PPROM group reached a significantly higher median amniotic fluid concentration of PTX3 compared with those in the midtrimester group (1.0ng/mL [0.36-121.0] versus 0.67ng/mL [0.4-2.8]; P=0.007). CONCLUSION: Histological chorioamnionitis is associated with a significant increase of amniotic fluid pentraxin 3 levels. Amniotic fluid pentraxin 3 appears to be a marker of intra-amniotic inflammation. PMID- 20022321 TI - Epidemiology of mixed urinary incontinence in China. AB - OBJECTIVES: To determine the prevalence and risk factors for mixed urinary incontinence (MUI) among Chinese women. METHOD: In this cross-sectional survey study we analyzed the answers to a modified Bristol Female Lower Urinary Tract Symptoms questionnaire provided by 19024 women older than 20 years and from 6 regions in China. RESULTS: The overall prevalence of MUI was 9.4%. The prevalence of MUI increased with age and reached 24.1% among women older than 70 years. Multivariable logistic regression analysis revealed that age, vaginal delivery, chronic constipation, pelvic organ prolapse, chronic pelvic pain, respiratory system disease, menstrual disorder, urinary system disease, alcohol consumption, pelvic surgery, obesity, and a low educational level were independent risk factors for MUI. CONCLUSION: We found age, vaginal delivery, and chronic constipation to be the main risk factors for MUI among adult women in China. PMID- 20022322 TI - The Chinese version of the Facts on Aging Quiz scale: reliability and validity assessment. AB - BACKGROUND: The use of health care services by older adults is increasing with the aging of the population. It is therefore essential that health care workers are current in their knowledge about gerontology; thus, educational programs regarding the care of elders are essential. Chinese-language assessments to measure knowledge about gerontology and aging and attitudes toward older adults are lacking. OBJECTIVE: This study was designed to measure the reliability and validity of a Chinese-language version of Palmore's facts on aging quiz (FAQ 1). METHOD: Cronbach's alpha, construct validity, content validity and convergent validity of the Chinese version of the FAQ 1 were assessed. A sample of 220 student nurses was recruited. Part of the sample (n=125; Group A) was comprised of experienced nurses who had returned to school for advanced degrees. The remaining 95 participants were first-year undergraduate students without prior work experience (Group B). The FAQ 1 was administered to all participants. The Chinese-language version of Kogan's Attitudes toward Older People (KAOP) was also administered to assess convergent validity of the FAQ 1. RESULTS: Cronbach's alpha of the FAQ was 0.68, content validity was 0.82, and construct validity and convergent validity were acceptable. Group A had more correct answers on each item than Group B, indicating that students with prior work experience with the elderly were more knowledgeable regarding older people and aging. CONCLUSIONS: Based on the results of the assessment, the Chinese version of FAQ 1 is a valid and reliable instrument for measuring students' or health care providers' knowledge about older people. PMID- 20022323 TI - Obovatol from Magnolia obovata inhibits vascular smooth muscle cell proliferation and intimal hyperplasia by inducing p21Cip1. AB - AIMS: Obovatol is isolated from Magnolia obovata leaves and this active component has various pharmacological properties such as anti-oxidant, anti-platelet, anti fungal and anti-inflammatory activities. In the present study, we investigated the inhibitory effects of obovatol on in vitro vascular smooth muscle cell (VSMC) proliferation and in vivo neointimal formation in a rat carotid artery injury model. METHODS AND RESULTS: Obovatol (1-5 microM) exerted concentration-dependent inhibition on platelet-derived growth factor (PDGF)-BB-induced rat VSMC proliferation, without exhibiting any cellular toxicity or apoptosis, as determined by cell count, [3H]thymidine incorporation and Annexin-V-binding analyses. Treatment with obovatol blocked the cell cycle in G1 phase by down regulating the expression of cyclins and CDKs, and selectively up-regulating the expression of p21Cip1, a well-known CDK inhibitor. Effects of perivascular delivery of obovatol were assessed 14 days after injury. The angiographic mean luminal diameters of the obovatol-treated groups (100 microg and 1 mg: 0.78+/ 0.06 and 0.77+/-0.07AU, respectively) were significantly larger than that of the control group (0.58+/-0.07AU). The obovatol-treated groups (100 microg and 1mg: 0.14+/-0.04 and 0.09+/-0.03 mm2, respectively) showed significant reduction in neointimal formation versus the control group (0.17+/-0.02 mm2). Immunohistochemical staining demonstrated strong expression of p21Cip1 in the neointima of the obovatol-treated groups. CONCLUSIONS: These data suggest that obovatol inhibits VSMC proliferation by perturbing cell cycle progression, possibly due to activation of p21Cip1 pathway. These results also show that obovatol may have potential as an anti-proliferative agent for the treatment of restenosis and atherosclerosis. PMID- 20022324 TI - Arterial stiffness predicts cardiovascular death independent of arterial thickness in a cohort of hemodialysis patients. AB - OBJECTIVE: Both arterial thickness and stiffness are predictors of cardiovascular disease (CVD). Although these arterial changes develop in parallel, no study has ever tested a hypothesis that arterial stiffness can predict mortality from CVD independent of arterial thickness. This study tested this possibility. METHODS: This was an observational cohort study in 423 hemodialysis patients (CKD stage 5D). We simultaneously measured intima-media thickness (CA-IMT) and stiffness parameter beta (CA-beta) by carotid ultrasonography at baseline, and the cohort was followed-up for a mean period of 70 months. RESULTS: During the follow-up, 216 all-cause deaths occurred including 124 deaths from CVD. Univariate analyses indicated both CA-IMT and CA-beta were significant predictors for CVD death. Kaplan-Meier analysis, in which the total subjects were divided into four groups by the medians of CA-IMT and CA-beta, showed that the hazards ratio (95% confidence interval) was 5.87 (3.43-10.05) for the group with higher CA IMT/higher CA-beta as compared to the group with lower CA-IMT/lower CA-beta. The hazards ratios for the group with lower CA-IMT/higher CA-beta (2.22, 1.16-4.25) and the group with higher CA-IMT/lower CA-beta (2.85, 1.52-5.33) were comparable. Multivariate Cox analysis revealed that both CA-IMT and CA-beta were independently predictive of CVD mortality even after adjustment for other relevant covariates. CONCLUSION: Increased arterial stiffness predicted cardiovascular mortality independent of arterial thickness in this cohort, implicating the distinct roles of stiffness and thickness of arterial wall in the pathogenesis of CVD. PMID- 20022325 TI - LDL and HDL subclasses in acute ischemic stroke: prediction of risk and short term mortality. AB - OBJECTIVE: Small, dense low-density lipoprotein (sdLDL) and small-sized high density lipoprotein (HDL) particles are established risk factors for ischemic heart disease. However, their clinical significance for acute ischemic stroke (AIS) is uncertain. This study evaluates associations of LDL and HDL particle sizes and subclasses with AIS risk and short-term mortality after AIS. METHODS: Two hundred AIS patients hospitalised for first-in-a-lifetime stroke and 162 apparently healthy controls were included in the study. LDL and HDL particles were separated by gradient gel electrophoresis and serum lipid parameters were measured by standard laboratory methods. Baseline characteristics of LDL and HDL particles were evaluated for the prediction of AIS and short-term mortality after AIS. RESULTS: AIS patients had significantly more LDL III and IVb, but less LDL I and II particles. They also had significantly smaller HDL size, more HDL 3a, 3b and 3c and less HDL 2b subclasses. The relative content of both sdLDL and small sized HDL particles was significantly increased in patients (P<0.001 and P<0.001, respectively). In addition, sdLDL was significantly higher in AIS fatalities (n=25) compared with survivors (n=175, P<0.05). Increased sdLDL was a significant predictor of AIS (OR=4.31; P<0.001) and in-hospital mortality after AIS (OR=5.50; P<0.05). The observed relationships persisted after adjustment for conventional risk factors. CONCLUSIONS: AIS is associated with adverse distributions of LDL and HDL subclasses. In addition, short-term mortality after AIS is associated with increased sdLDL particles. Our results indicate that sdLDL is an independent predictor of both AIS onset and consecutive short-term mortality. PMID- 20022326 TI - Uncovering chaotic structure in mechanomyography signals of fatigue biceps brachii muscle. AB - The mechanomyography (MMG) signal reflects mechanical properties of limb muscles that undergo complex phenomena in different functional states. We undertook the study of the chaotic nature of MMG signals by referring to recent developments in the field of nonlinear dynamics. MMG signals were measured from the biceps brachii muscle of 5 subjects during fatigue of isometric contraction at 80% maximal voluntary contraction (MVC) level. Deterministic chaotic character was detected in all data by using the Volterra-Wiener-Korenberg model and noise titration approach. The noise limit, a power indicator of the chaos of fatigue MMG signals, was 22.20+/-8.73. Furthermore, we studied the nonlinear dynamic features of MMG signals by computing their correlation dimension D(2), which was 3.35+/-0.36 across subjects. These results indicate that MMG is a high dimensional chaotic signal and support the use of the theory of nonlinear dynamics for analysis and modeling of fatigue MMG signals. PMID- 20022327 TI - Application of charged single isomer derivatives of cyclodextrins in capillary electrophoresis for chiral analysis. AB - The review focuses on the role of ionic or ionisable single isomer derivatives (SIDs) of cyclodextrins on the separation of chiral analytes in capillary electrophoresis (CE), covering the period since the year 2000. The advantages of using pure compounds are discussed, as well as the ways to optimise the separations in the context of a rational approach to these techniques. Specific attention is paid to the modulation of the selector-analyte interaction. The advantage due to a detailed knowledge of equilibria occurring in solution during the CE run is underlined, particularly in the case of the presence of metal complexes, as occurs in chiral ligand exchange capillary electrophoresis (CLECE). PMID- 20022328 TI - Optimization of a pressurized liquid extraction method by experimental design methodologies for the determination of fluoroquinolone residues in infant foods by liquid chromatography. AB - In the present study, we have developed a method based on pressurized liquid extraction (PLE) and liquid chromatography with fluorescence detection (LC-FLD) for the determination of residues of fluoroquinolones (FQs) in infant food products. PLE extraction has been optimized by the application of experimental design methodologies. Initially, a fractional factorial design (FFD) was used to screen the significance of four extraction parameters: solvent composition, temperature, pressure and number of cycles. The most significant factors, identified by ANOVA analysis, were the solvent composition, temperature and pressure, which were further optimized with the aid of a face centred design (FCD) and the desirability function. The optimized operating PLE conditions were as follows: ACN/o-phosphoric acid 50mM pH 3.0 (80:20, v/v), 80 degrees C, 2000psi and three extraction cycles of 5min. Under these conditions, recoveries of the target FQs varied between 69% and 107% with RSDs below 9%. The whole method was validated according to the Commission Decision 2002/657/EC guidelines. The proposed method has been successfully applied to the analysis of different infant food products bought in local supermarkets and pharmacies. The results showed the presence of residues of enrofloxacin in a non-compliant baby food sample corresponding to a chicken-based formulation, which were also confirmed and quantified by LC-MS/MS analysis. PMID- 20022329 TI - Liquid chromatography-atmospheric pressure photoionization tandem mass spectrometry for analysis of 36 halogenated flame retardants in fish. AB - A comprehensive, sensitive and high-throughput liquid chromatography-atmospheric pressure photoionization tandem mass spectrometry (LC-APPI-MS/MS) method has been developed for analysis of 36 halogenated flame retardants (HFRs). Under the optimized LC conditions, all of the HFRs eluted from the LC column within 14min, while maintaining good chromatographic separation for the isomers. Introduction of the pre-heated dopant to the APPI source decreased the background noise fivefold, which enhanced sensitivity. An empirical equation was proposed to describe the relation between the ion intensity and dopant flow. The excellent on column instrument detection limits averaged 4.7pg, which was similar to the sensitivity offered by gas chromatography-high-resolution mass spectrometry (GC HRMS). This method was used to analyze a series of fish samples. Good agreement was found between the results for PBDEs from LC-APPI-MS/MS and GC-HRMS. PMID- 20022330 TI - Chiral Separations - In honour of Prof. Volker Schurig. PMID- 20022331 TI - Optimisation of ambient and high temperature asymmetric flow field-flow fractionation with dual/multi-angle light scattering and infrared/refractive index detection. AB - Asymmetric flow field-flow fractionation (AF4) enables to analyse polymers with very high molar masses under mild conditions in comparison to size exclusion chromatography (SEC). Conventionally, membranes for AF4 are made from cellulose. Recently, a novel ceramic membrane has been developed which can withstand high temperatures above 130 degrees C and chlorinated organic solvents, thus making it possible to characterise semicrystalline polyolefins by HT-AF4. Two ceramic membranes and one cellulose membrane were compared with regard to their quality of molar mass separation and the loss of the polymer material through the pores. Separating polystyrene standards as model compounds at different cross-flow gradients the complex relationship between cross-flow velocity, separation efficiency, the molar mass and peak broadening could be elucidated in detail. Moreover, the dependence of signal quality and reproducibility on sample concentration and mass loading was investigated because the evaluation of the obtained fractograms substantially depends on the signal intensities. Finally, the performance of the whole system was tested at high temperature by separating PE reference materials of high molar mass. PMID- 20022332 TI - Direct identification of phenolic constituents in Boldo Folium (Peumus boldus Mol.) infusions by high-performance liquid chromatography with diode array detection and electrospray ionization tandem mass spectrometry. AB - A very simple and direct method was developed for the qualitative analysis of polyphenols in boldo (Peumus boldus Mol., Monimiaceae) leaves infusions by high performance liquid chromatography with diode array detection (HPLC-DAD) and electrospray ionization tandem mass spectrometry (HPLC-MS(n)). The phenolic constituents identified in infusions of the crude drug Boldo Folium were mainly proanthocyanidins and flavonol glycosides. In the infusions, 41 compounds were detected in male and 43 compounds in female leaf samples, respectively. Nine quercetin glycosides, eight kaempferol derivatives, nine isorhamnetin glycosides, three phenolic acids, one caffeoylquinic acid glycoside and twenty one proanthocyanidins were identified by HPLC-DAD and ESI-MS for the first time in the crude drug. Isorhamnetin glucosyl-di-rhamnoside was the most abundant flavonol glycoside in the male boldo sample, whereas isorhamnetin di-glucosyl-di rhamnoside was the main phenolic compound in female boldo leaves infusion. The results suggest that the medicinal properties reported for this popular infusion should be attributed not only to the presence of catechin and boldine but also to several phenolic compounds with known antioxidant activity. The HPLC fingerprint obtained can be useful in the authentication of the crude drug Boldo Folium as well as for qualitative analysis and differentiation of plant populations in the tree distribution range. PMID- 20022333 TI - Preparative enantioseparation of (+/-)-N-(3,4-cis-3-decyl-1,2,3,4 tetrahydrophenanthren-4-yl)-3,5-dinitrobenzamide by centrifugal partition chromatography. AB - The racemic compound (+/-)-N-(3,4-cis-3-decyl-1,2,3,4-tetrahydrophenanthren-4-yl) 3,5-dinitrobenzamide ((+/-)-1), an analogue of increased lipophilicity of the chiral selector (CS) contained in the Whelk-O HPLC chiral stationary phase (CSP) has been resolved into its enantiomers by applying centrifugal partition chromatography (CPC). Considering the known enantioselectivity of the Whelk-O CS for naproxen, and the reciprocity concept in enantioseparation, (S)-naproxen related compounds were tested as CSs. In the search for an adequate solvent system, the partition behaviour of the two solutes, CS and racemate, has been studied using several biphasic solvent mixtures. The optimal CS concentration and sample loading capacity were determined in the chosen solvent system. The search for an appropriate CS and solvent system, the scale-up and optimization of the enantiomer separation by CPC, as well as the rationale for the unexpected elution order of enantiomers, are here described. The comparison of the preparative CPC separation achieved with that in HPLC, using a CSP containing an analogous CS, resulted favourable to the former in terms of loading capacity, solvent consumption and throughput. PMID- 20022334 TI - Analysis of special surfactants by comprehensive two-dimensional gas chromatography coupled to time-of-flight mass spectrometry. AB - Multidimensional gas-chromatographic analyses of olesochemically based nonionic, anionic and several cationic surfactants in industrial cleaners are demonstrated. Comprehensive two-dimensional gas chromatography coupled with time-of-flight mass spectrometry allows the simultaneous determination of fatty alcohols, fatty alcohol sulphates and alkyl polyglucosides. In addition, the determination of fatty alcohol ethoxylates up to C(10)EO(8) (highest degree of ethoxylation) and C(18)EO(5) (longest C-chain at an ethoxylation degree of five) and the analysis of fatty alcohol alkoxylates that contain ethoxy (EO) and propoxy (PO) groups could be realized. Because of decomposition in the injector and a weak EI fragmentation, cationic surfactants such as alkyl benzyl dimethyl ammonium chloride could also be identified by their characteristic fragments. Thermogravimetric analyses confirmed that the temperature in a normal GC injector is not high enough to cause thermal decomposition of esterquats. However, we could demonstrate that a modified silylation procedure forms decomposition products of esterquats in the GC injector which are detectable by GCxGC-(TOF)MS and allows the identification of such GC-atypical analytes. PMID- 20022335 TI - Microfabricated planar glass gas chromatography with photoionization detection. AB - We report the development of a microfabricated gas chromatography system suitable for the separation of volatile organic compounds (VOCs) and compatible with use as a portable measurement device. Hydrofluoric acid etching of 95x95mm Schott B270 wafers has been used to give symmetrical hemi-spherical channels within a glass substrate. Two matching glass plates were subsequently cold bonded with the channels aligned; the flatness of the glass surfaces resulted in strong bonding through van der Waals forces. The device comprised gas fluidic interconnections, injection zone and 7.5 and 1.4m long, 320microm internal diameter capillaries. Optical microscopy confirmed the capillaries to have fully circular channel profiles. Direct column heating and cooling could be achieved using a combination of resistive heaters and Peltier devices. The low thermal conductivity of glass allowed for multiple uniform temperature zones to be achieved within a single glass chip. Temperature control over the range 10-200 degrees C was achieved with peak power demand of approximately 25W. The 7.5m capillary column was static coated with a 2microm film of non-polar dimethylpolysiloxane stationary phase. A standard FID and a modified lightweight 100mW photoionization detector (PID) were coupled to the column and performance tested with gas mixtures of monoaromatic and monoterpene species at the parts per million concentration level. The low power GC-PID device showed good performance for a small set of VOCs and sub ng detection sensitivity to monoaromatics. PMID- 20022336 TI - Time dependence of nucleation and growth of silver nanoparticles generated by sugar reduction in micellar media. AB - Synthesis of silver nanoparticles in homogeneous aqueous solutions of the precursors silver nitrate and three saccharides (two mono saccharides i.e. glucose and fructose and one disaccharide such as sucrose) has been performed. These Ag-nanoparticles can be derivatized by other surface treatments, as required. The anionic surfactant SDS was tested to investigate its effect on the dependence of size, growth rate, photoluminescence (PL) emission and polydispersity of the nanoparticles. The time evolution of UV-vis absorbance suggested that nucleation and growth rate markedly vary in a first order fashion w.r.t. Ag(+) salt concentration. The differences in the reducing ability of the saccharides were discussed with reference to their structure dependent adsorption behavior onto the particles. Changes in UV-vis, PL and FT-IR spectra during nucleation and growth of the nanoparticles were used to establish plausible mechanisms for the adsorption of surfactant on the particle surface, so as to restrict the growth. The results revealed a simple and easy strategy for synthesizing metal nanoparticles with well controlled shapes, sizes and structures. PMID- 20022337 TI - Controlling the morphology of electrospray-generated PLGA microparticles for drug delivery. AB - We developed a well-controlled method to generate PLGA microparticles of different morphologies using the electrospray drying route. By judiciously selecting polymer molecular weight, concentration, and solution flow rate, we can control the order in which polymer entanglements and Coulomb fission occur in the droplets and their relative importance, and subsequently govern the morphology of the resulting polymer particles. We show that spherical, monodisperse particles are generated when sufficiently strong polymer entanglements set in the evaporating droplets before they undergo any Coulomb fission. On the other hand, tailed and elongated particles are obtained if the Coulomb fission occurs first and if the droplets/particles are sufficiently evaporated to freeze in their irregular shape. Strictly spherical particles are unachievable for polymer solutions below a critical concentration, because the onset of Coulomb fission always sets in prior to the development of a sufficiently entangled polymer network. An extension of a simple model, originally used to determine the onset of electrospinning of polymer solutions, adequately predicts when non-spherical particles are produced. We conclude by demonstrating the scale-up of this approach to the synthesis of polymer particles using a compact, microfabricated, multiplexed electrospray system, which would make it suitable for practical applications. PMID- 20022338 TI - The growth hormone-insulin-like growth factor axis in glycogen storage disease type 1: evidence of different growth patterns and insulin-like growth factor levels in patients with glycogen storage disease type 1a and 1b. AB - OBJECTIVES: To investigate the growth hormone (GH)-insulin-like growth factor (IGF) system in patients with glycogen storage disease type 1 (GSD1). STUDY DESIGN: This was a prospective, case-control study. Ten patients with GSD1a and 7 patients with GSD1b who were given dietary treatment and 34 sex-, age-, body mass index-, and pubertal stage-matched control subjects entered the study. Auxological parameters were correlated with circulating GH, either at basal or after growth hormone releasing hormone plus arginine test, insulin-like growth factors (IGF-I and IGF-II), and anti-pituitary antibodies (APA). RESULTS: Short stature was detected in 10.0% of patients with GSD1a, 42.9% of patients with GSD1b (P = .02), and none of the control subjects. Serum IGF-I levels were lower in patients with GSD1b (P = .0001). An impaired GH secretion was found in 40% of patients with GSD1a (P = .008), 57.1% of patients with GSD1b (P = .006), and none of the control subjects. Short stature was demonstrated in 3 of 4 patients with GSD1b and GH deficiency. The prevalence of APA was significantly higher in patients with GSD1b than in patients with GSD1a (P = .02) and control subjects (P = .03). The GH response to the provocative test inversely correlated with the presence of APA (P = .003). Compared with levels in control subjects, serum IGF II and insulin levels were higher in both groups of patients, in whom IGF-II levels directly correlated with height SD scores (P = .003). CONCLUSION: Patients with GSD1a have an impaired GH secretion associated with reference range serum IGF-I levels and normal stature, whereas in patients with GSD1b, the impaired GH secretion, probably because of the presence of APA, was associated with reduced IGF-I levels and increased prevalence of short stature. The increased IGF-II levels, probably caused by increased insulin levels, in patients with GSD1 are presumably responsible for the improved growth pattern observed in patients receiving strict dietary treatment. PMID- 20022339 TI - Roles of parental sleep/wake patterns, socioeconomic status, and daytime activities in the sleep/wake patterns of children. AB - OBJECTIVES: To determine sleep/wake patterns of primary school children and their correlates. STUDY DESIGN: A total of 4470 sets of mother-father-child community based trios were recruited in this study. We constructed 3 integrated models with structural equation modeling to predict sleep/wake patterns of children (bedtime, wakeup time, and time in bed [TIB]). RESULTS: Our best-fitting models explained 40% to 71% variances of various sleep/wake patterns of the children, which were influenced by a web of interactive factors including school start time, parental sleep/wake patterns, sociodemographics, and daytime activities. The strongest predictor of various sleep/wake patterns was school start time. Higher socioeconomic status would shorten TIB of both children and parents, but through different pathways (by advancing wakeup time and delaying bedtime in children but by delaying bedtime in parents). Media use and homework shortened TIB of children, while leisure extracurricular activities and later school start time lengthened it. The age and sex effects on sleep/wake patterns, at least in part, were mediated by daytime activities. Daytime activities of children also influenced their parental sleep/wake patterns, especially their maternal one. A consistent pattern of stronger mother-child than father-child associations were found in various sleep/wake patterns. CONCLUSIONS: There was a complex and interactive relationship among school schedule, parental sleep/wake patterns, socioeconomic status, and daytime activities in determining the sleep/wake patterns of children. These findings have important clinical implications for the management of childhood sleep/wake habits and problems. PMID- 20022340 TI - Biomarkers of hypercoagulability and inflammation in childhood-onset arterial ischemic stroke. AB - OBJECTIVE: To test the hypothesis that acute elevations of biomarkers of hypercoagulability and inflammation are common in children with arterial ischemic stroke (AIS), particularly among etiologic subtypes that carry an increased risk of recurrent stroke. STUDY DESIGN: In this prospective/retrospective institutional-based cohort study of acute childhood-onset AIS (n = 50) conducted between 2005 and 2009, D-dimer, factor VIII (FVIII) activity, C-reactive protein (CRP), and erythrocyte sedimentation rate (ESR) were serially evaluated at the time of clinical blood sampling. Patients were classified by stroke subtype as cardioembolic, moyamoya, non-moyamoya arteriopathy, or other. RESULTS: Both D dimer and CRP were frequently elevated in acute childhood-onset AIS and exhibited a decreasing trend with time. Acute D-dimer levels were significantly higher in cardioembolic AIS compared with noncardioembolic AIS (median, 2.04 microg/mL [range 0.54-4.54 microg/mL] vs 0.32 microg/mL [0.22-3.18 microg/mL]; P = .002). At an optimal threshold of > or = 0.50 microg/mL, the sensitivity and specificity of D-dimer for cardioembolic subtype were 78% and 79%, respectively. CONCLUSIONS: Our findings identify D-dimer and CRP as candidate biomarkers for etiology and prognosis in childhood-onset AIS. Further studies should investigate the role of these and other biomarkers of hypercoagulability and inflammation in childhood onset AIS. PMID- 20022341 TI - Lung volume and ventilation inhomogeneity in preterm infants at 15-18 months corrected age. AB - OBJECTIVE: To assess whether lung volume and ventilation inhomogeneity in preterm infants at 15-18 months corrected age, and the change in these outcomes from the newborn period to 15-18 months corrected age, depend on gestational age (GA) at birth and the severity of neonatal lung disease. STUDY DESIGN: Preterm (GA range, 23-32 weeks) and term healthy control infants were studied in quiet sedated sleep at 15-18 months corrected age by multiple breath washout with 5% sulfur hexafluoride using an ultrasonic flowmeter. Valid measurements were obtained from 58 infants. Multivariate and multilevel regression was used to analyze outcomes. RESULTS: Functional residual capacity (FRC), lung clearance index, and first and second to zeroeth moment ratios were calculated. After accounting for body size at test, FRC at follow-up, and the increase in FRC from the newborn period to 15 18 months corrected age were positively associated with GA and negatively associated with the duration of endotracheal ventilation. Indices of ventilation inhomogeneity were unaltered by GA and the duration of endotracheal ventilation. CONCLUSIONS: In very preterm infants, GA and the duration of endotracheal ventilation are independently associated with reduced lung volume and lung growth during infancy, although the effect size of these findings is small. PMID- 20022342 TI - Duration of fever affects the likelihood of a positive bag urinalysis or catheter culture in young children. AB - OBJECTIVE: To test the hypothesis that there will be a clinically significant rise in the proportion of positive bag urinalyses and catheter cultures in young children with increasing duration of fever. STUDY DESIGN: This was a prospective cohort study of 818 infants and children age 3-36 months presenting to a tertiary care emergency department with documented fever without source. Following the documentation of fever from < 1 to > or = 5 days, bag specimens were collected for urinalysis. The primary outcome was the yield of positive bag dipsticks by day, defined as positive for nitrates or more than trace leukocyte esterase. The secondary outcome was positive catheter cultures on each day of fever. RESULTS: Positive bag urinalyses increased with duration of fever: 14.8% (35/237) on day 1 versus 26.4% (43/163) on day 3 (relative risk [RR] = 1.8; 95% confidence interval [CI] = 1.2-2.7; P = .004). Positive catheter cultures increased in the same fashion: 4.8% (11/229) on day 1 versus 12.6% (20/159) on day 3 (RR = 2.6; 95% CI = 1.3-5.3; P = .005). CONCLUSIONS: The yield of positive bag urinalyses and catheter cultures increased significantly in children with fever of 3 days or longer duration. PMID- 20022344 TI - Effects of short-term stress-like cortisol on cerebral metabolism: a proton magnetic resonance spectroscopy study at 3.0 T. AB - Preclinical and clinical studies suggest that glucocorticoids disturb cognitive performance and neuronal integrity. MR spectroscopy studies have tried to track these effects and correlate long-term effects of glucocorticoids with concentration changes of cerebral metabolites. However, a systematic spectroscopic study on short-term exposure to corticosteroids, in a dosage sufficient to impair memory performance, is lacking. Thus, it is not known when glucocorticoid effects become visible at 1H-MRS in vivo. Therefore, employing localized 1H-MRS at 3T we quantitatively investigated the effects of a 4-day cortisol exposure (160 mg/d) on brain metabolites in vivo in a double blind and placebo-controlled cross-over study of 21 healthy subjects. Spectroscopic measurements were performed in four different brain regions, including the posterior cingulate gyrus, right frontal white matter, left and right anterior hippocampus using the PRESS method. Our results show, after 4 days of cortisol intake and despite a nearly 3-fold increase in serum cortisol concentration, for none of the investigated metabolites a significant corticosteroid-induced concentration change. Our results show that contrary to reported long-term effects, short-term hydrocortisone exposure at a stress-like dosage does not lead to changes of prominent cerebral metabolites, including N-acetylaspartate, creatine and phosphocreatine, choline-containing metabolites, myo-inositol and glutamate. PMID- 20022343 TI - Prevalence of intracardiac shunting in children with sickle cell disease and stroke. AB - OBJECTIVE: To determine the prevalence of potential intracardiac shunts, including patent foramen ovale (PFO), in children with sickle cell disease (SCD) and stroke. STUDY DESIGN: We performed a transthoracic echocardiogram (TTE) on 40 children with SCD (39 with hemoglobin SS and 1 with sickle-beta0 thalassemia) and earlier stroke (overt stroke in 30, silent infarction in 10). We compared 3 TTE techniques: conventional 2-dimensional imaging, color Doppler ultrasound, and intravenous agitated saline contrast injection for the detection of intracardiac shunts. We also evaluated the clinical, laboratory, and radiographic findings of the children with and without shunts. RESULTS: We identified PFO or other potential intracardiac shunts in 10 of 40 children with SCD and earlier stroke (25%; 95% CI, 11.6-38.4). With contrasted TTE, we failed to detect potential shunts in 2 children. In a comparison group of 60 children with stroke but without SCD, retrospective review of clinical echocardiograms identified PFO in 7 of 60 (11.7%; 95% CI, 3.6-19.8). Clinical features significantly associated with the presence of intracardiac shunts were stroke in the setting of vaso-occlusive crisis (P = .026) and headache at stroke onset (P = .014). CONCLUSION: One quarter of children with SCD and stroke have potential intracardiac shunts. A combination of echocardiographic techniques is required for optimal shunt detection. Intracardiac shunting could be a risk factor for stroke in children with SCD because they are predisposed to thrombosis and elevations of right heart pressure, which could promote paradoxical embolization across an intracardiac shunt. PMID- 20022345 TI - Venlafaxine and mirtazapine treatment lowers serum concentrations of dehydroepiandrosterone-sulfate in depressed patients remitting during the course of treatment. AB - OBJECTIVES: The adrenal androgen dehydroepiandrosterone-sulfate (DHEA-S) seems to be involved in the pathophysiology of depression, although its precise role in the etiology and remission of depression remains unclear. In the present study we intended to examine possible differential effects of venlafaxine and mirtazapine in a randomised open trial with regard to DHEA-S serum concentrations in patients suffering from major depressive episode compared to healthy controls. METHODS: We assessed DHEA-S concentrations both at baseline and after a 4-week treatment period in 70 depressed patients (n=33 for venlafaxine and n=37 for mirtazapine) and 33 matched healthy controls. RESULTS: We describe the decrease of DHEA-S levels in depressive patients who remitted after treatment with both venlafaxine or mirtazapine. Patients without remission of depression did not show a significant decline in DHEA-S concentrations. CONCLUSIONS: Our results suggest an effect of treatment outcome upon DHEA-S concentrations rather than a direct drug effect. The change of plasma DHEA-S levels as a marker of treatment-response of depression warrant further investigation. PMID- 20022346 TI - Denaturing gradient gel electrophoresis shows that bacterial communities change with mid-ocean ballast water exchange. AB - Ships carry ballast water for better stability and to control trim. However, the discharge of ballast water near ports is known to transport invasive species from one coastal area to another. The exchange of ballast water on the high seas is supposed to reduce such invasions of exotic species. In this study, we used denaturing gradient gel electrophoresis (DGGE) to analyze the composition of the bacterial community in ballast water before and after such a mid-ocean exchange, and we also measured total bacterial counts. Our findings confirmed that the ballast water was replaced by the mid-ocean exchange, as indicated by the marked change in the composition of the bacterial community. There was also a significant decrease in bacterial abundance after the mid-ocean exchange. Finally, our findings support the incubation hypothesis, because the composition of the bacterial communities changed over time within the same ballast water. PMID- 20022347 TI - Phenolic compounds and their anti-oxidative properties and protein kinase inhibition from the Chinese mangrove plant Laguncularia racemosa. AB - Phenolic compounds, named integracin D (1), (7'R, 8'S, 8S)-8-hydroxyisoguaiacin (3), (2R, 3R) pinobanksin-3-caffeoylate (5) and threo-8S-7 methoxysyringylglycerol (6), respectively, were isolated from the Chinese mangrove plant Laguncularia racemosa (L) Gaertn. f. (Combretaceae), together with 23 known phenolic metabolites. Their structures were elucidated on the basis of extensive spectroscopic analyses including that of IR, UV, MS, CD, 1D and 2D NMR spectra as well as by comparison with literature data. Compound 5 showed significant anti-oxidative activity in the DPPH and TEAC free-radical-scavenging assays, while several of the phenolic compounds were tested for protein kinase inhibitory activity in an assay involving 24 different human tumor related protein kinases. Compounds 5, 7, and 23 showed potential inhibition with IC(50) values between 2.2 and 3.6microg/mL toward individual kinases. The ellagic acid derivatives were tested for insecticidal activity. PMID- 20022348 TI - The influence of late pregnancy and lactation on bone metabolism in mares. AB - Pregnancy and lactation are periods of significant influence on bone metabolism that has not been investigated in equines. To examine the influence of late pregnancy and lactation on bone metabolism in mares, the changes in the blood serum/plasma total calcium (Ca), inorganic phosphates (Pi), pyridinoline (Pyd) and 17beta-estradiol (E2) concentration and the bone alkaline phosphatase (BAP) activity were investigated. The samples were taken from 11 mares on 60+/-10 and 20+/-10 days before foaling, and 20+/-10 and 60+/-10 days after foaling. The concentration/activity of Ca, Pi, Pyd and BAP increased significantly in early lactation, but the Pyd than decreased in the 4th period. A significant correlation was observed between the E2 and bone metabolism parameters. The results indicate low maintenance of normocalcaemia with reduced bone synthesis in late pregnancy and prove the role of estradiol in bone metabolism in mares during pregnancy and lactation. PMID- 20022349 TI - Augmentation of CCL17 and CCL28 gene expression by TNF-alpha, IL-1beta, or IFN gamma in cultured canine keratinocytes. AB - Keratinocytes produce inflammatory mediators that are involved in the pathogenesis of skin disorders such as atopic dermatitis (AD). In particular, the CC chemokines, thymus and activation regulated chemokine (TARC)/CCL17 and mucosae associated epithelial chemokine (MEC)/CCL28 are considered to play an important role in the lesional infiltration of lymphocytes in canine AD. However, there have been no reports on the regulatory mechanisms of CCL17 and CCL28 transcription in canine keratinocytes. In this study, we investigated whether CCL17 and CCL28 transcription in cultured keratinocytes is induced by TNF-alpha, IL-1beta, or IFN-gamma. It was found that CCL17 mRNA transcription is augmented by TNF-alpha only, whereas the CCL28 mRNA level could be increased by TNF-alpha, IL-1beta, or IFN-gamma. The present study suggests that pro-inflammatory cytokines are important inducing factors for the production of CCL17 and CCL28 in the lesional skin of dogs with AD. PMID- 20022350 TI - Wastewater for agriculture: A reuse-oriented planning model and its application in peri-urban China. AB - The benefits of Integrated Water Resources Management (IWRM) are widely known but its recommendations remain thinly implemented. Designing wastewater treatment plants for reuse in irrigation is a particularly underutilized IWRM opportunity that could potentially increase agricultural yields, conserve surface water, offset chemical fertilizer demand, and reduce the costs of wastewater treatment by eliminating nutrient removal processes. This paper presents a novel planning model, consisting of a reuse-centric performance assessment and optimization model to help design wastewater treatment plants for reuse in agriculture. The performance assessment and optimization model are described, and their coupled application is demonstrated in the peri-urban district of Pixian, China. Based on the results of the performance assessment, two reuse scenarios are evaluated: wastewater to supplement business as usual (BAU) irrigation, and wastewater to replace BAU irrigation. The results indicate that wastewater supplementation could increase profits by $20 million (M) annually; alternatively, wastewater replacement could conserve 35Mm(3) of water in local rivers each year. PMID- 20022351 TI - Comprehensive life cycle inventories of alternative wastewater treatment systems. AB - Over recent decades, the environmental regulations on wastewater treatment plants (WWTP) have trended towards increasingly stringent nutrient removal requirements for the protection of local waterways. However, such regulations typically ignore other environmental impacts that might accompany apparent improvements to the WWTP. This paper quantitatively defines the life cycle inventory of resources consumed and emissions produced in ten different wastewater treatment scenarios (covering six process configurations and nine treatment standards). The inventory results indicate that infrastructure resources, operational energy, direct greenhouse gas (GHG) emissions and chemical consumption generally increase with increasing nitrogen removal, especially at discharge standards of total nitrogen <5 mgN L(-1). Similarly, infrastructure resources and chemical consumption increase sharply with increasing phosphorus removal, but operational energy and direct GHG emissions are largely unaffected. These trends represent a trade-off of negative environmental impacts against improved local receiving water quality. However, increased phosphorus removal in WWTPs also represents an opportunity for increased resource recovery and reuse via biosolids applied to agricultural land. This study highlights that where biosolids displace synthetic fertilisers, a negative environmental trade-off may also occur by increasing the heavy metals discharged to soil. Proper analysis of these positive and negative environmental trade-offs requires further life cycle impact assessment and an inherently subjective weighting of competing environmental costs and benefits. PMID- 20022352 TI - Enhancement of biogas production by addition of hemicellulolytic bacteria immobilised on activated zeolite. AB - Biogas from agricultural biomass and residues is a valuable source of renewable energy. However, recalcitrant plant cell structures represent a barrier in the fermentative biodegradation process in single- and two-stage reactors. Therefore, approaches concerning a more efficient de-polymerisation of cellulose and hemicellulose to monomeric sugars are required amongst others in order to optimise the fermentation efficiency and to increase methane yields. Here we show a new strategy for the enhancement of biogas production from hemicellulose-rich substrates. Hemicellulolytic populations from a common biogas fermenter consortium were successively enriched in batch-cultures using a synthetic medium containing xylan powder as single carbon source under anaerobic mesophilic conditions. Enriched hemicellulolytic bacteria were immobilised on trace metal activated zeolite to ensure a stable storage and easy application. Xylanase activity increased continuously during subsequent enrichment cycles by up to 162%. In batch-culture experiments we were able to observe an increase of methane by 53% compared to controls without additionally introduced microorganisms immobilised on zeolite. Specific enrichment of hemicellulolytic bacteria during the process was confirmed by using single strand conformation polymorphism (SSCP) analysis based on amplification of the eubacterial 16S rDNA fragments. Using sequence analysis conspicuous bands from SSCP patterns could be identified as belonging to the groups Bacteroides sp., Azospira oryzae (Dechlorosoma sp.) as well as to a wide spectrum of diverse species within the order of Clostridiales (Firmicutes). PMID- 20022353 TI - Arsenic sorption on TiO2 nanoparticles: size and crystallinity effects. AB - Single solute As (III) and As (V) sorption on nano-sized amorphous and crystalline TiO(2) was investigated to determine: size and crystallinity effects on arsenic sorption capacities, possible As (III) oxidation, and the nature of surface complexes. Amorphous and crystalline nanoparticles were prepared using sol-gel synthesis techniques. For amorphous TiO(2), solute pH in the range of 4-9 had a profound impact on only As (V) sorption. As (III) and As (V) sorption isotherms indicated that sorption capacities of the different TiO(2) polymorphs were dependent on the sorption site density, surface area (particle size) and crystalline structure. When normalized to surface area, As (III) surface coverage on the TiO(2) surface remained almost constant for particles between 5 and 20 nm. However, As (V) surface coverage increased with the degree of crystallinity. X ray absorption spectroscopic analysis provided evidence of partial As (III) oxidation on amorphous TiO(2) rather than crystalline TiO(2). The data also indicated that As (III) and As (V) form binuclear bidentate inner-sphere complexes with amorphous TiO(2) at neutral pH. PMID- 20022354 TI - Polychlorinated biphenyls and organochlorine pesticides in European eel (Anguilla anguilla) from the Garigliano River (Campania region, Italy). AB - Measuring organochlorine compounds in muscle tissue of European eels from the Garigliano River in Campania (Italy), overall PCBs emerged as the most abundant pollutants, followed by DDTs, Dieldrin and HCB. Target PCBs, IUPAC nos. 118, 138, 153 and 180, were the dominant congeners accounting for 64.2% of total PCBs. Among OCPs, p,p'-DDE was detected in all eels, always with higher concentration levels than other OCPs; p,p'-DDT was frequently detected, about 93.3% of the sample. The high and statistically significant correlations between concentration and length as well as weight of eels suggest that the organochlorine compounds concentrations tend to increase with the size and consequently with the age. Concentrations of DDTs and PCBs detected were similar to those reported in studies relative to France, UK and Sweden. As regards toxicological risk for human health, in general OCPs residual levels were below the limits established for fish and aquatic products. Conversely, the concentrations of PCBs exceeded the limit set by the EU for terrestrial foods. Our results imply that OCPs and PCBs are still important persistent chemical contaminants in Campania freshwaters, although their manufacture and use are banned or highly restricted. PMID- 20022355 TI - Behaviour of (14)C-sulfadiazine and (14)C-difloxacin during manure storage. AB - The persistence of sulfadiazine, difloxacin, and their metabolites has been investigated in stored manure. The manure collected from sulfadiazine ((14)C-SDZ) and difloxacin ((14)C-DIF) treated pigs contained N-acetylsulfadiazine (Ac-SDZ), 4-hydroxy-SDZ (4-OH-SDZ), and sarafloxacin (SARA) as the main metabolites, respectively along with their parent compounds. Manures were stored separately at 10 degrees C and 20 degrees C at various moisture levels. About 96-99% of the radioactivity remained in extractable parent compounds and their metabolites after 150d of storage. The formation of non-extractable residue and the rate of mineralization were both negligible in manure containing SDZ and DIF. During storage SDZ concentration increased as a result of the deacetylation of Ac-SDZ, whose concentration decreased proportionally. Hence the environmental effects may be underestimated if the parent compound alone is considered for environmental risk assessment. About 11% and 14% of 4-OH-SDZ were lost after 20 and 40d of storage; thereafter its concentration increased relatively, highlighting hydroxylation of SDZ. DIF degraded very slowly (7% loss after 150d) during the storage of manure; in contrast the concentration of SARA decreased rapidly (72 90% loss after 150d). Dilution of manure and storage at higher temperatures for a reasonable period of time enhanced the rate of reactions of SDZ, DIF and their related metabolites. PMID- 20022356 TI - Factor XII Ofunato: Lys346Asn mutation associated with blood coagulation factor XII deficiency causes impaired secretion through a proteasome-mediated degradation. AB - INTRODUCTION: Congenital blood coagulation factor XII (FXII) deficiency is a rare coagulation disease and an autosomal recessive trait. It is found by chance in many cases. We identified a novel mutation (Lys346Asn) in the FXII gene of a patient with FXII deficiency, designated as Factor XII Ofunato. METHODS: The proband was a 75-year-old Japanese woman with a prolonged activated partial thromboplastin time (52.8s). The FXII activity and antigen were greatly reduced (activity, 5%; antigen, 4.5%). We analyzed FXII gene of this patient using a direct sequencing method and characterized mutant FXII through in vitro expression studies. RESULTS: Sequence analysis of the FXII gene revealed a G-->C point mutation at nucleotide 9845, resulting in Lys346 (AAG)-->Asn (AAC) replacement in the catalytic domain. Expression studies in Chinese hamster ovary cells demonstrated that mutant FXII (346N-FXII) showed a lower level of accumulation in the cells than wild-type. Secretion of 346N-FXII was greatly reduced in culture medium. We also investigated mRNA expression levels of wild type and 346N-FXII in transfected cells using quantitative RT-PCR. Both mRNA expressions were equivalent levels. Pulse-chase experiments showed that 346N-FXII was extensively degraded intracellularly compared to wild-type. Using membrane permeable inhibitors, we observed that degradation occurred in the pre-Golgi compartment and that proteasome apparently plays a central role in this process. CONCLUSIONS: These results show that most 346N-FXII is degraded intracellularly through endoplasmic reticulum-associated degradation as the protein quality control system, resulting in an insufficient secretion phenotype. PMID- 20022357 TI - Circulating microparticles are elevated in carriers of factor V Leiden. AB - INTRODUCTION: Microparticles (MP) are small membrane bound cellular particles that play an important role in thrombosis. This study was carried out to evaluate if increased numbers or procoagulant potential of circulating MP contribute to the heterogeneity in occurrence of thrombosis in heterozygotes carrying Factor V Leiden (FVL) mutation. METHODS: Levels of circulating platelet (CD41a), endothelial (CD62e) as well as leukocyte (CD45) derived MP from 45 FVL heterozygous individuals were enumerated by flow cytometry and compared with normal controls. Functional studies included enzyme linked immunoassay based prothrombinase activity (ELISA) and modified dilute Russell Viper venom test (DRVVT). RESULTS: Circulating MP were significantly higher in the FVL cohort compared to the controls (median=2100 vs. 1508 MP/microl, respectively p=0.0021).All subsets of MP (platelet, endothelial and leukocyte) were significantly elevated in the FVL group, the most striking disparity seen in the number of CD45 positive leukocyte MP. Despite the differences in the number of MP between the controls and FVL cohorts, there was no significant difference in the prothrombinase activity recorded by the ELISA (2.0 vs 2.4 PS equivalents; p=0.7374) or clotting time assessed by the DRVVT (47 vs 46 sec, p=0.8118). When the FVL cohort was considered alone there was no significant difference in MP parameters between FVL subjects with or without a history of thrombosis. CONCLUSIONS: This is the first study on circulating MP levels in subjects who are heterozygote for factor V Leiden. We report that circulating platelet and leukocyte MP are elevated in carriers of this mutation and may be important contributors to risk of thrombosis. PMID- 20022358 TI - A novel splice site mutation in intron C of PROS1 leads to markedly reduced mutant mRNA level, absence of thrombin-sensitive region, and impaired secretion and cofactor activity of mutant protein S. AB - Protein S (PS) is a member of the vitamin K-dependent protein family containing similar gamma-carboxyglutamic acid (Gla) domains, although only PS has a thrombin sensitive region (TSR), which is located between the Gla domain and the first epidermal growth factor-like domain. In this study, a novel PROS1 mutation was identified at the last nucleotide in intron C (c.260-1G>A) in a patient suffering from recurrent deep vein thrombosis associated with PS deficiency. To investigate the molecular mechanisms of PS deficiency caused by the novel PROS1 mutation, we characterized the mutant mRNA, and the secretion and function of the mutant PS molecule associated with the mutation. RT-PCR was used to detect the aberrant mRNA in the patient's platelets, the amount of which was markedly reduced and lacked the region corresponding to exon 4 coding the TSR of the PS molecule. The recombinant mutant PS lacking the TSR (TSR-lack PS) showed a markedly reduced transient expression/secretion level, 37.9% of that of wild-type (WT) PS. Activated protein C (APC) cofactor activity assay showed that TSR-lack PS had no cofactor activity. Moreover, binding assays of monoclonal antibodies recognizing the PS Gla domain and the Gla residues indicated that the bindings of TSR-lack PS to both of these antibodies were clearly weaker than those of WT PS. These findings suggest that the novel mutation leading to the absence of the TSR not only affected the secretion of mutant PS, but was also responsible for impairment of the Gla domain conformation required for the gamma-carboxylation to express APC cofactor activity. PMID- 20022359 TI - Adjuvant treatment with concomitant radiotherapy and chemotherapy in high-risk endometrial cancer: a clinical experience. AB - OBJECTIVE: The concurrent use of radiotherapy (RT) and chemotherapy (CT) as adjuvant treatment after surgery in high-risk endometrial cancer has been generally considered cautiously. Recently some of us have reported preliminary data on the efficacy and tolerability of concomitant CT and RT. In this paper, we update our experience. METHODS: A total of 47 patients aged >18 years and <80 years with histological diagnosis of high-risk endometrial endometrioid carcinomas entered the study. Inclusion criteria were stages IC G3, IIB, IIIA (patients with positive washing without other unfavourable prognostic factors were omitted), IIIB and IIIC. The radiation plan consisted of a total dose of 50.4 Gy, given in five fractions per week (1.8 Gy: daily dose) for 6 weeks. Paclitaxel (P) at a dose of 60 mg/m(2) was infused intravenously in 250 mL of normal saline for 1 h once weekly during RT for 5 weeks. Three further cycles of Paclitaxel, at a dose of 80 mg/m(2), have been given weekly at the end of RT. RESULTS: There was no life-threatening toxicity. The overall 5-year relapse-free survival was 81.8% (95% CI, 65.2-90.9). The 5-year percent overall disease specific survival was 88.4% (95% CI, 71.1-95.6). CONCLUSIONS: These results, based on a larger series, support our previous data: Paclitaxel plus RT may represent an effective and well-tolerated treatment in high-risk endometrial cancer patients. PMID- 20022360 TI - Response to M. Pluta: "Less radical surgery than radical hysterectomy in early stage cervical cancer--a pilot study"; Gynecol Oncol 2009;113:181-184. PMID- 20022361 TI - PGFM (13,14-dihydro-15-keto-PGF(2alpha)) in pregnant and pseudo-pregnant Iberian lynx: a new noninvasive pregnancy marker for felid species. AB - In mammals, uterine and placental prostaglandin F(2alpha) is involved in the regulation of reproduction-related processes such as embryonic development, initiation of parturition, and resumption of ovarian activity. Prostaglandin F(2alpha) (PGF(2alpha)) is rapidly metabolized to its plasma metabolite PGFM (13,14-dihydro-15-keto-PGF(2alpha)), which has also been detected in urine. Therefore, the current study aimed to develop and validate an efficient, quick, and inexpensive enzyme immunoassay (EIA) for PGFM estimation in urine of the Iberian lynx (Lynx pardinus) for pregnancy monitoring and for differentiation between pregnancy and pseudo-pregnancy. Urine samples collected from captive Iberian lynx (11 pregnant and 4 pseudo-pregnant cycles) were subjected directly to a PGFM EIA. The assay was validated for parallelism, precision, and stability of urinary PGFM. In addition, high-performance liquid chromatography (HPLC) immunograms and liquid chromatography-mass spectrometry (LCMS) were performed to identify PGFM within urine samples. Urinary PGFM levels before mating and after parturition were about 1.5 ng/mL. After Day 20 postmating, both pregnant and pseudo-pregnant females showed slight increase of hormone levels; in pseudo pregnant females, this elevation did not exceed 7 ng/mL. A significant increase in pregnant females was observed after Day 45 postmating; urinary PGFM increased from 10 ng/mL at Day 45 toward a peak of 46.0+/-19.3 ng/mL around parturition. First results show that PGFM is detectable in feces as well and follows similar courses as shown for urine. In conclusion, the presented and validated PGFM assay is an easy and reliable method for noninvasive pregnancy diagnosis in the Iberian lynx (and probably other felids) if applied approximately 20 d prior parturition in pure urine or fecal extracts. High PGFM levels in urine or fecal samples may allow a pregnancy diagnosis without knowledge of mating time, making the PGFM test applicable to free-ranging animals. PMID- 20022362 TI - Oxytocin, vasopressin, prostaglandin F(2alpha), luteinizing hormone, testosterone, estrone sulfate, and cortisol plasma concentrations after sexual stimulation in stallions. AB - This experiment was designed to determine the effects of sexual stimulation on plasma concentrations of oxytocin (OT), vasopressin (VP), 15-ketodihydro PGF(2alpha) (PG-metabolite), luteinizing hormone (LH), testosterone (T), estrone sulfate (ES), and cortisol (C) in stallions. Semen samples were collected from 14 light horse stallions (Equus caballus) of proven fertility using a Missouri model artificial vagina. Blood samples were collected at 15, 12, 9, 6, and 3 min before estrous mare exposure, at erection, at ejaculation, and at 3, 6, and 9 min after ejaculation. Afterwards, blood sampling was performed every 10 min for the following 60 min. Sexual activity determined an increase in plasma concentrations of OT, VP, C, PG-metabolite, and ES and caused no changes in LH and T concentrations. The finding of a negative correlation between C and VP at erection, and between C and T before erection and at the time of erection, could be explained by a possible inhibitory role exerted by C in the mechanism of sexual arousal described for men. PMID- 20022363 TI - Is Doublesynch protocol a new alternative for timed artificial insemination in anestrous dairy cows. AB - This is the very first report that suggests high pregnancy rates can be obtained with use of the Doublesynch protocol in anestrous dairy cows. Recently, a new synchronization method has been developed (Doublesynch) that resulted in synchronized ovulations both after the first and second gonadotropin-releasing hormone (GnRH) treatments. It was suggested that this protocol has the potential to increase the pregnancy rates in primiparous dairy cows. The aim of the current study was to confirm the success of the Doublesynch protocol and further to investigate the effect of this method on pregnancy rates in anestrous cows. Lactating primiparous Holstein (Bos taurus) cows (n=165) between 60 and 172 d postpartum were monitored twice with 10-d intervals (on Days -10 and 0) by ultrasonography, and blood samples were collected. Cows were classified as anestrous if both blood samples had progesterone (P4) concentration <1 ng/mL and as cyclic if at least one of the two samples had P4 concentration >or=1 ng/mL. Cyclic cows were classified again as cyclic-high P4 (having an active corpus luteum) if the second blood samples had P4 concentrations >or=1 ng/mL and as cyclic-low P4 if P4 concentrations were <1 ng/mL on Day 0. Then, the cows classified as anestrous (n=51), cyclic-high P4 (n=63), or cyclic-low P4 (n=51) were put into two treatment groups (Ovsynch or Doublesynch) randomly to establish six groups. Cows in the Ovsynch group were administered a GnRH (lecirelin 50 microg, im) on Day 0, PGF (Prostaglandin F2 alpha, D-cloprostenol 0.150 mg, im) on Day 7, and a second dose of GnRH 48 h later. Cows in the Doublesynch group were administered a PGF on Day 0, GnRH on Day 2, a second PGF on Day 9, and a second GnRH on Day 11. Timed artificial insemination (TAI) was performed 16 to 20 h after the second GnRH in both treatment groups. Pregnancy diagnosis was conducted (by ultrasonography) 45+/-5 d after TAI. In anestrous cows and those with high and low progesterone concentration at treatment onset, Doublesynch treatment led to markedly increased pregnancy rates with respect to Ovsynch treatment (P<0.05). On the overall analysis of data, it was revealed that the Doublesynch method increased pregnancy rates by 43 percentage units (29.8% vs. 72.8%, P<0.0001) in relation to Ovsynch. Pregnancy rates of cows having small, medium, or large follicles at the day of second GnRH administration were similar in the Doublesynch group (70.4%, 85.2%, and 63.0%, respectively; P>0.05), whereas pregnancy rates reduced dramatically as follicle size increased in the Ovsynch group, particularly in cows with follicles greater than 16 mm (45.5%, 28.1%, and 5.3%, respectively; P<0.05). Our results confirm and support observations that the Doublesynch protocol increases the pregnancy rates in postpartum primiparous cows as reported previously. Our data also demonstrate that the Doublesynch method increases the pregnancy rates in anestrous cows. Thus, these data suggest that the Doublesynch protocol can be used to obtain satisfactory pregnancy rates after TAI in both anestrous and cycling primiparous dairy cows regardless of stage of estrous cycle. PMID- 20022364 TI - Family cultural socialization practices and ethnic identity in college-going emerging adults. AB - We examined how family cultural socialization related to the ethnic identity of Asian American, Latino, White, and Mixed-Ethnic emerging adults (N=225). Greater family cultural socialization was related to greater ethnic identity exploration and commitment. Ethnic minority students reported higher levels of family cultural socialization and ethnic identity compared to White students. The family cultural socialization-ethnic identity link was more pronounced for females compared to males, and for White compared to ethnic minority students. The findings highlight the importance of the family for identity development beyond adolescence. PMID- 20022365 TI - The osteogenic effect of electrosprayed nanoscale collagen/calcium phosphate coatings on titanium. AB - For orthopedic and dental implants, the ultimate goal is to obtain a life-long secure anchoring of the implant in the native surrounding bone. To this end, nanoscale calcium phosphate (CaP) and collagen-CaP (col-CaP) composite coatings have been successfully deposited using the electrospray deposition (ESD) technique. In order to study to what extent the thickness of these coatings can be reduced without losing coating osteogenic properties, we have characterized the mechanical and biological coating properties using tape tests (ASTM D-3359) and in vitro cell culture experiments, respectively. Co-deposition of collagen significantly improved coating adhesive and cohesive strength, resulting in a remarkably high coating retention of up to 97% for coating thicknesses below 100 nm. In vitro cell culture experiments showed that electrosprayed CaP and col-CaP composite coatings enhanced osteoblast differentiation, leading to improved mineral deposition. This effect was most pronounced upon co-deposition of collagen with CaP, and these coatings displayed osteogenic effects even for a coating thickness of below 100 nm. PMID- 20022366 TI - In vitro engineering of human ear-shaped cartilage assisted with CAD/CAM technology. AB - Due to the lack of appropriate scaffolds, the in vitro engineering of cartilage tissue with a sophisticated structure, such as a human ear, remains a great challenge. Although polyglycolic acid (PGA) has become one of the most successful scaffolds for cartilage regeneration, how to overcome its limitations in achieving desirable mechanical strength and accurate control over shape remains an unsolved problem. In this study, the mechanical strength of PGA scaffold was enhanced by coating with polylactic acid (PLA). The content of PLA was optimized by balancing the scaffold's biocompatibility and mechanical strength. The PLA/PGA scaffold was then fabricated into a human ear-shape mirror-symmetrical to a normal ear by pressing the scaffold in the ear negative molds, which were fabricated by the computer aided design and manufacturing (CAD/CAM) technique according to the CT scan data from the normal ear. The ear-shaped scaffold reached a similarity level of over 97% compared to the positive ear mold by the shape analysis using a 3D laser scan system. Most importantly, after chondrocyte seeding, the constructs largely retained the original shape during culture with a similarity level of over 84%. Furthermore, the constructs formed ear-shaped cartilage-like tissues at 12 weeks, which revealed a tissue structure with abundant cartilage extracellular matrices and mature lacuna. Additionally, the ear-shaped cartilage at 12 weeks also exhibited fine elasticity and good mechanical strength. These results may provide a useful strategy for reconstructing cartilage tissue with complicated shapes such as a human ear by an in vitro engineering approach. PMID- 20022367 TI - The use of carboxymethylcellulose gel to increase non-viral gene transfer in mouse airways. AB - We have assessed whether viscoelastic gels known to inhibit mucociliary clearance can increase lipid-mediated gene transfer. Methylcellulose or carboxymethylcellulose (0.25-1.5%) was mixed with complexes of the cationic lipid GL67A and plasmids encoding luciferase and perfused onto the nasal epithelium of mice. Survival after perfusion with 1% CMC or 1% MC was 90 and 100%, respectively. In contrast 1.5% CMC was uniformly lethal likely due to the viscous solution blocking the airways. Perfusion with 0.5% CMC containing lipid/DNA complexes reproducibly increased gene expression by approximately 3-fold (n=16, p<0.05). Given this benefit, likely related to increased duration of contact, we also assessed the effect of prolonging contact time of the liposome/DNA complexes by delivering our standard 80 microg DNA dose over either approximately 22 or 60 min of perfusion. This independently increased gene transfer by 6-fold (n=8, p<0.05) and could be further enhanced by the addition of 0.5% CMC, leading to an overall 25-fold enhancement (n=8, p<0.001) in gene expression. As a result of these interventions CFTR transgene mRNA transgene levels were increased several logs above background. Interestingly, this did not lead to correction of the ion transport defects in the nasal epithelium of cystic fibrosis mice nor for immunohistochemical quantification of CFTR expression. To assess if 0.5% CMC also increased gene transfer in the mouse lung, we used whole body nebulisation chambers. CMC was nebulised for 1h immediately before, or simultaneously with GL67A/pCIKLux. The former did not increase gene transfer, whereas co administration significantly increased gene transfer by 4-fold (p<0.0001, n=18). This study suggests that contact time of non-viral gene transfer agents is a key factor for gene delivery, and suggests two methods which may be translatable for use in man. PMID- 20022368 TI - Effect of delivery of MMP inhibitors from PDMS as a model IOL material on PCO markers. AB - Posterior capsule opacification (PCO) or secondary cataract formation, following intraocular lens implantation, is a significant complication affecting an estimated 28% of cataract patients. Matrix metalloproteinases (MMPs) have been demonstrated to play a role in the formation of anterior subcapsular cataracts and it has been shown that the presence of MMP inhibitors (MMPI) decreases subcapsular cataract formation ex vivo. Since the mechanisms responsible for anterior subcapsular cataract formation and posterior capsule opacification are similar, it is reasonable to suggest that MMP inhibitors may also mitigate PCO. One of the most effective ways of delivering the inhibitors may be from the implanted intraocular lens (IOL) material itself. In the current work, delivery of three different MMP inhibitors from silicone rubber as a model IOL material was examined. Loading methods were developed which allowed continuous release of active MMPI for periods of over 5 months in some cases. Reduced migration rates were observed in human lens epithelial cells in vitro, suggesting that an effect on PCO may be possible. While further studies are necessary to tune the systems to achieve the desired rates of release, this work demonstrates that delivery of MMPI from silicone IOL materials has the potential to decrease the incidence of PCO. PMID- 20022369 TI - Cyclodextrin-based device coatings for affinity-based release of antibiotics. AB - Cyclodextrin-based hydrogels were synthesized to create robust networks with tunable mechanical properties capable of serving as device coatings. The CD networks were able to swell and load drug in aqueous and organic solvents. The rheological properties of the swollen gels were investigated using stress and frequency sweeps, with both demonstrating high storage modulus, indicating strong elastic gels. The ability of the gels to swell in numerous solvents allowed for the separate loading and release of different antibiotic drug molecules with varying hydrophilicities. Based on FTIR and TGA studies, each drug was found to form an inclusion complex with CD. For comparison, dextran gels were prepared similarly. As expected for affinity-based mechanisms, the release of drugs from the CD-based gels was slower than diffusion-based release from the dextran gels, and could be sustained for more than 200 h. Coating potential was tested by coating two different medical devices: metal screws and polymer meshes. The meshes were characterized by SEM, revealing that CD-based coatings resulted in a uniform thin film, whereas the dextran gels only partly coated the device and showed delamination. Considerably longer bactericidal activity against Staphylococcus aureus was observed for both the CD hydrogels and coatings, as compared to dextran-based ones. The slow, sustained, affinity-based release of antibiotics from the CD-based networks reflects their potential as a delivery platform. PMID- 20022370 TI - The role of sterilization in the cytocompatibility of titania nanotubes. AB - Titiania nanotubes have large potential in medical implant applications but their tissue compatibility is still controversial. Since the sterilization methods may impact the biocompatibility of titania nanotubes and be the source of the controversy, we investigate the influence of three commonly used sterilization methods, autoclaving, ultraviolet irradiation and ethanol immersion, on the cytocompatibility of titania nanotubes. Two titania nanostructures, namely nanonets with an average pore diameter of 25 nm and nanotubes with an average diameter of 80 nm, are used in this study. The results show that the sterilization methods significantly affect the cytocompatibility of these titania surfaces. UV and ethanol sterilization give rise to a higher surface free energy inducing higher initial cell adhesion and proliferation compared to autoclaving, whereas UV irradiation produces the best cell functions including adhesion, proliferation, as well as differentiation represented by related gene expressions. The cytocompatibility results obtained from the nanoscale surfaces are compared to those acquired from the polished surface demonstrating the significant effects. Our results suggest that the sterilization process plays an important role in the observed cytocompatibility of titania nanotubes and may be the reason for the controversial results so far. UV sterilization is found to be the best method from the viewpoint of surface contamination elimination. PMID- 20022371 TI - A biodegradable, injectable, gel system based on MPEG-b-(PCL-ran-PLLA) diblock copolymers with an adjustable therapeutic window. AB - In situ-forming gel systems have drawn increasing attention for their potential use in a variety of biomedical applications. Here, we examined an in situ-forming gel system comprised of MPEG-b-PCL and MPEG-b-(PCL-ran-PLLA) diblock copolymers with different PLLA contents (0-10 mol%) in the PCL segment. The crystalline region of the PCL-ran-PLLA segment decreased with increasing PLLA content. The MPEG-b-(PCL-ran-PLLA) diblock copolymer solutions were liquid at room temperature and only MPEG-b-(PCL-ran-PLLA) diblock copolymer solutions with a PLLA content < or = 5 mol% in the PCL segment showed a sol-to-gel transition as the temperature was increased. The viscosity change associated with sol-to-gel phase transition depended on the PLLA content in the PCL segment. A MPEG-b-PCL diblock copolymer solution incubated in vitro showed increasing viscosity without degradation, whereas the viscosity of MPEG-b-(PCL-ran-PLLA) diblock copolymer solutions continuously and sharply decreased with increasing PLLA content in the PCL segment. As the amount of PLLA increased, the size of in vivo-formed MPEG-b-(PCL ran-PLLA) gels after initial injection tended to gradually decrease because of hydrolytic degradation of the PLLA in the PCL-ran-PLLA segment. An immunohistochemical examination showed that in vivo MPEG-b-(PCL-ran-PLLA) diblock copolymer gels provoked only a modest inflammatory response. Collectively, our results show that the MPEG-b-(PCL-ran-PLLA) diblock copolymer gel described here could serve as a minimally invasive, therapeutic, in situ-forming gel system that offers an experimental window adjustable from a few weeks to a few months. PMID- 20022372 TI - Cellular expression of the monocarboxylate transporter (MCT) family in the placenta of mice. AB - Lactate plays an important role as an alternative energy substrate, especially in conditions with a decreased utility of glucose. Proton-coupled monocarboxylate transporters (MCTs) are essential for the transport of lactate, ketone bodies, and other monocarboxylates through the plasma membrane and may contribute to the net transport of lactate through the placental barrier. The present study examined the expression profile and subcellular localization of MCTs in the mouse placenta. An in situ hybridization survey of all MCT subtypes detected intense mRNA expressions of MCT1, MCT4, and MCT9 as well as GLUT1 in the placenta from gestational day 11.5. The expression of MCT mRNAs decreased in the intensity at the end of gestation in contrast to a consistently intense expression of GLUT1 mRNA. Immunohistochemically, MCT1 and MCT4 showed a polarized localization on the maternal side and fetal side of the two cell-layered syncytiotrophoblast, respectively. The membrane-oriented localization of MCTs was supported by the coexistence of CD147 which recruits MCT to the plasma membrane. However, the subcellular arrangement of MCT1 and MCT4 along the trophoblastic cell membrane was completely opposite of that in the human placenta. Although we cannot exactly explain the reversed localization of MCTs between human and murine placentas, it may be related to differences between humans and mice in the origin of lactate and its utilization by fetuses. PMID- 20022373 TI - Large granular lymphocytic (LGL) leukemia in an adult with Down syndrome (47,XX,+21). PMID- 20022374 TI - Recurrent involvement of heterochromatic regions in multiple myeloma-a multicolor FISH study. AB - Chromosome aberrations are important prognostic markers in multiple myeloma (MM), but their identification may be hampered by complexity of the karyotypes. Using multicolor fluorescence in situ hybridization (mFISH), we found cryptic aberrations in 7 of 10 patients with a complex karyotype. Moreover, in addition to typical aberrations involving 1q, 13q, 14q and 17p and structural aberrations in chromosomes 1, 6, 9 and 19, (iso)dicentric chromosomes and whole-arm translocations were detected. These chromosome aberrations were generated by breaks in heterochromatic regions indicating an increased breakage of these regions, which may predispose to the generation of chromosome aberrations in multiple myeloma. PMID- 20022375 TI - Neutrophilic eccrine hidradenitis associated with decitabine. PMID- 20022376 TI - Pollution by psychoactive pharmaceuticals in the Rivers of Madrid metropolitan area (Spain). AB - There are a number of reports in the literature which describe the occurrence of so-called emerging pollutants, such as pharmaceuticals, in surface water. Few of these studies have reported values from large cities in relatively arid areas, such as in Spain. The persistence of some pharmaceuticals to usual wastewater treatments allows their discharge into surface waters. It is increasingly evident that mental health problems are of special concern for public health since psychiatric drugs are used in large amounts. Compared to other countries, Spain has a high pharmaceutical consumption rate, and Madrid metropolitan area is one of the most densely populated areas in Europe. The aim of the present study was to determine the presence of different psychoactive pharmaceuticals and metabolites in the main rivers of Madrid metropolitan area: Jarama, Manzanares, Guadarrama, Henares and Tajo. Sampling was done downstream of ten sewage treatment plants (STP) discharging into these rivers. Control points upstream of STPs discharge points were also sampled. Pharmaceutical compounds and metabolites for analysis were selected according to human consumption and prescription rates in Madrid, and the availability of valid techniques for detection. We observed residues of the antidepressants fluoxetine (80% of the sampling sites), citalopram (60%) and venlafaxine (100%), the anxiolytics nordiazepam (90%), oxazepam (80%) and 7-aminoflunitrazepam (10%) and the anticonvulsant carabamazepine (70%). Measured concentrations equalled or exceeded those reported for other geographical areas, although there is a pronounced lack of information for the anxiolytics and venlafaxine. This is of special concern given that Wyeth Ayerst's venlafaxine, Effexor, was the 10th greatest selling pharmaceutical worldwide in 2006. We conclude that the origin of pharmaceutical pollution in the rivers of Madrid is mainly the discharge of sewage treatment plants in Madrid's metropolitan area and a comprehensive monitoring program should be implemented. PMID- 20022377 TI - Pairwise decomposition of residue interaction energies of single chain Fv with HIV-1 p17 epitope variants. AB - Computational assisted modeling was carried out to investigate the importance of specific residues in the binding site of scFv. In this study, scFv against HIV-1 epitope at the C-terminal on p17 (scFv anti-p17) was used as a candidate molecule for evaluating the method. The wild-type p17 and its nine natural mutants were docked with scFv anti-p17. Potential mean force (PMF) scores predicted the most favorable binding interaction, and the correlation agreed well with the corresponding activity data from the peptide based ELISA. In the interaction with solvent molecules, the 3D structures of scFv anti-p17 and selected peptide epitopes were further investigated by molecular dynamics (MDs) simulation with the AMBER 9 program. Post-processing of the snapshot at equilibrium was performed to evaluate the binding free energy and pairwise decomposition or residue-based energy calculation of complexes in solution using the Molecular Mechanics Poisson Boltzmann Surface Area (MM-PBSA) protocol. Our results demonstrated that the specific residues located in the complementary determining regions (CDRs) of scFv anti-p17, MET100, LYS101, ASN169, HIS228, and LEU229, play a crucial role in the effective binding interaction with the absolute relative decomposed energy more than 2.00 kcal/mol in comparison to the original substrate. PMID- 20022378 TI - Calmodulin inhibition of E2A stops expression of surrogate light chains of the pre-B-cell receptor and CD19. AB - To create antibody diversity, B lymphocyte development is characterized by the ordered rearrangement of first immunoglobulin (Ig) heavy chain gene segments and then Ig light-chain gene segments. Early in B-cell development, expression of a pre-B-cell receptor (pre-BCR) composed of membrane-bound Ig heavy chain protein associated with surrogate light-chain (SLC) proteins serves as a critical checkpoint that monitors for functional heavy chain rearrangement. Signaling from the pre-BCR induces clonal expansion, but it also turns off transcription of the genes for the SLC proteins lambda5 and VpreB, which limits this proliferation. Here we show that signaling from the pre-BCR rapidly down-regulates lambda5 and VpreB and also the co-receptor CD19 in primary pre-B-cells. We show that calcium (Ca(2+)) signaling is essential for this silencing of the SLC and CD19 genes. The SLC genes are activated by the E2A transcription factor, and we show that E2A is required for pre-BCR-mediated regulation of the genes. E2A mutated in its binding site for the Ca(2+) sensor protein calmodulin, and thus with calmodulin-resistant DNA binding, makes lambda5, VpreB and CD19 expression resistant to the inhibition following pre-BCR activation. Thus, Ca(2+) down-regulates SLC and CD19 gene expression upon pre-BCR activation through inhibition of E2A by Ca(2+)/calmodulin. PMID- 20022379 TI - Balb/c EGFP mice are tolerant against immunization utilizing recombinant adenoviral-based vectors encoding EGFP: a novel model for the study of tolerance mechanisms and vaccine efficacy. AB - Enhanced green fluorescent protein (EGFP) is a marker gene product which is readily detectable using the techniques of fluorescence microscopy, flow cytometry, or macroscopic imaging. Previous studies have demonstrated the immunogenicity of EGFP in Balb/c mice, identifying an immunodominant H2-K(d) restricted CTL epitope. To model immunological tolerance and vaccine efficiency against self-antigens, we generated a stable transgenic BALB/c mouse expressing EGFP (Balb/c EGFP) through back-crossing C57Bl/6-TG(ACTbEGFP)10sb more than ten times with Balb/c wildtype (wt) mice. High level EGFP expression was detected in the skin and heart, whereas low level expression was observed in the kidney, liver, gut, lung, and spleen. To characterize the immune reactivity to self antigen, we immunized Balb/c EGFP and Balb/c wt mice with recombinant adenoviral based vectors encoding EGFP (Ad-EGFP) or beta-galactosidase (Ad-betagal) as a control. Immunization utilizing the Ad-betagal vector expressing 'foreign' antigen induced robust humoral and cellular transgene-specific immunity, whereas Balb/c EGFP mice presented no reactivity following Ad-EGFP immunization against the 'self-antigen' EGFP. These findings describe the creation of a transgenic mouse line tolerant against the common protein marker EGFP, providing a novel system for the evaluation of methods of tolerance disruption and vaccine efficacy. PMID- 20022381 TI - Binding of the complement inhibitor C4b-binding protein to Lyme disease Borreliae. AB - The Lyme disease spirochetes, Borrelia burgdorferi sensu stricto, Borrelia afzelii and Borrelia garinii, are tick-borne pathogens that can cause chronic disseminated infections. To survive in the human host borreliae need to evade the immune system. It is already well known that B. burgdorferi ss. and B. afzelii bind the complement (C) alternative pathway inhibitor factor H from serum using OspE and CRASP-1/Bba68 proteins to escape C attack. In the presence of natural antibodies and in chronic infections, when specific antibodies develop, borreliae have to protect themselves from antibody-induced classical pathway C attack. In this study we demonstrate binding of the classical pathway inhibitor, C4b-binding protein (C4bp), to three genospecies of B. burgdorferi sensu lato. Binding was strongest to B. garinii, which has been found to be the weakest factor H binder. The bacteria bound both purified (125)I-labeled C4bp and C4bp from serum. Unlabeled C4bp competed for binding with (125)I-C4bp, whereas BSA had no effect. Binding was salt-sensitive and inhibited by C4b and partially by heparin. C4bp maintained its cofactor activity for factor I in cleaving C4b when bound to the bacterial surface. Ligand blotting analysis of whole cell lysates and fractionated outer cell membranes of the bacteria suggested one major receptor of approximately 43 kDa (P43) for C4bp in B. garinii and B. burgdorferi sensu stricto. Binding of C4bp may thus allow Lyme disease borreliae to escape activation of the classical C pathway and allow chronic infections in humans even in the presence of specific antibodies. PMID- 20022380 TI - Endothelial P2Y2 receptor regulates LPS-induced neutrophil transendothelial migration in vitro. AB - Previous studies showed that P2 receptors are involved in neutrophil migration via stimulation of chemokine release and by facilitating chemoattractant gradient sensing. Here, we have investigated whether these receptors are involved in LPS induced neutrophil transendothelial migration (TEM) using a Boyden chamber where neutrophils migrated through a layer of lipopolysaccharide (LPS)-stimulated human umbilical vein endothelial cells (HUVECs). In line with a role of P2 receptors, neutrophil TEM was inhibited by the P2 receptor antagonists suramin and reactive blue 2 (RB-2) acting on the basolateral, but not luminal, HUVECs' P2 receptors. HUVECs express P2Y(1), P2Y(2), P2Y(4), P2Y(6) and P2Y(11). The involvement of P2Y(4) was unlikely as this receptor is insensitive to suramin while P2Y(1), P2Y(6) and P2Y(11) were excluded with available selective antagonists, leaving P2Y(2) as the only candidate. Indeed, the P2Y(2) knockdown in HUVECs inhibited neutrophil TEM compared to control HUVECs transfected with scrambled siRNA. Moreover, UTP, a P2Y(2) ligand, markedly potentiated LPS-induced TEM. Interestingly, IL-8 and ICAM-1 had a modest effect on neutrophil TEM in this 3h assay which was significantly diminished by the inhibition of Rho kinase in HUVECs with Y27632. In summary, endothelial P2Y(2) receptors control the early LPS-induced neutrophil TEM in vitro via Rho kinase activation. PMID- 20022382 TI - Obsessive-compulsive symptom dimensions in the general population: results from an epidemiological study in six European countries. AB - BACKGROUND: The prevalence of obsessive-compulsive symptom dimensions and their sociodemographic and psychopathological correlates at the population level are unknown. METHOD: Obsessive-compulsive symptom dimensions and mental disorders were assessed with the Composite International Diagnostic Interview 3.0 in a random subsample (n=2804) of individuals participating in a cross-sectional survey of the adult general population of six European countries. RESULTS: The lifetime prevalence of any obsessive-compulsive symptom dimension was 13%. Harm/Checking was the most prevalent dimension (8%) followed by Somatic obsessions (5%) and Symmetry/Ordering (3%). Females were more likely to have symptoms in Contamination/Cleaning (OR=3, 95%CI=1.06-8.51) and Somatic obsessions (OR=1.88, 95%CI=1.05-3.37). All symptom dimensions were associated with an increased risk of most mental (but not physical) disorders. There were some differences in prevalence between countries. LIMITATIONS: The interference associated with each symptom dimension could not be assessed. Few direct data are available on the validity of the CIDI to assess obsessive-compulsive symptom dimensions. CONCLUSIONS: Obsessive-compulsive symptom dimensions are relatively frequent in the general population. Their sociodemographic and psychopathological correlates may be slightly different in clinical and community samples. They are associated with an increased risk of most mental disorders. PMID- 20022383 TI - Symptoms of psychosis in anorexia and bulimia nervosa. AB - Despite evidence from case series, the comorbidity of eating disorders with psychosis is less investigated than their comorbidity with anxiety and mood disorders. We investigated the occurrence of symptoms of psychosis in 112 female patients diagnosed with DSM-IV eating disorders (anorexia nervosa=61, bulimia nervosa=51) and 631 high school girls in the same health district as the patients: the items of the SCL-90R symptom dimensions "paranoid ideation" and "psychoticism" were specifically examined. No case of co-morbid schizophrenia was observed among patients. Compared with controls, the patients with anorexia nervosa were more likely to endorse the item "Never feeling close to another person"; the patients with bulimia nervosa were more likely to endorse the item "Feeling others are to blame for your troubles". Both groups of patients were more likely than controls to endorse the item "Idea that something is wrong with your mind". The students who were identified by the EAT and the BITE as being "at risk" for eating disorders were more likely to assign their body a causative role in their problems. Symptoms of psychosis can be observed in patients with eating disorders, but these could be better explained within the psychopathology of the disorders rather than by assuming a link with schizophrenia. PMID- 20022384 TI - Principal domains of behavioral psychopathology identified by the Bipolar Inventory of Signs and Symptoms Scale (BISS). AB - Current symptom rating scales and diagnostic categories for bipolar disorder (BD) do not provide dimensional profiles of the types of behavior disturbed in this complex disorder. To overcome these limitations we identified the principal domains of behavioral symptomatology in bipolar individuals, including all mood states, and used a more comprehensive rating scale for BD: the Bipolar Inventory of Signs and Symptoms Scale (BISS). A total of 246 patients with BD (196 with BD type I, and 50 with BD type II) were interviewed using the BISS. Exploratory factor analysis was performed on the BISS results using the maximum likelihood factor extraction method, followed by oblique rotation of the extracted factor loadings. We determined the strength of relationships between factor scores using the Pearson correlation coefficient. The following five factors were extracted: mania, depression, irritability, anxiety and psychosis. Anxiety was significantly correlated with depression and irritability. The mania factor score was only weakly associated with the other four factors. The domains of the BISS capture both the historical categories of depression and mania, plus irritability, psychosis, and an additional principal domain, anxiety. Despite the common occurrence of anxiety in BD, it has not been identified in most prior factor analyses, in part due to limited coverage of anxiety symptoms in the source scales. PMID- 20022385 TI - Socioemotional deficits associated with obsessive-compulsive symptomatology. AB - Increasing emphasis has been placed on the role of socioemotional functioning in models of obsessive-compulsive disorder (OCD). The present study investigated whether OCD symptoms were associated with capacity for theory of mind (ToM) and basic affect recognition. Non-clinical volunteers (N=204) completed self report measures of OCD and general psychopathology, in addition to behavioral measures of ToM and affect recognition. The results indicated that higher OCD symptoms were associated with reduced ToM, as well as reduced accuracy decoding the specific emotion of disgust. Importantly, these relationships could not be attributed to other, more general features of psychopathology. The findings of the current study therefore further our understanding of how the processing and interpretation of social and emotional information is affected in the context of OCD symptomatology, and are discussed in relation to neuropsychological models of OCD. PMID- 20022386 TI - Canine CXCL7 and its functional expression in dendritic cells undergoing maturation. AB - Many cells, including leucocytes and stromal cells, express CXCL7, a member of the CXC chemokine family, also known as platelet basic protein. CXCL7 is a potent chemoattractant and activator of neutrophil function. Dendritic cells (DCs) play a pivotal role in antigen processing and presentation. Very little information is available on the ability of DCs to recruit neutrophils by producing chemokines. In this work, we have cloned canine CXCL7. Based on the predicted gene sequence and using the 3'RACE technique, the full-length gene was amplified from LPS treated canine peripheral blood mononuclear cells. The cloned cDNA sequence consisted of 357 nucleotides and encoded a 118 amino acid protein, including a 38 amino acid signal peptide. The use of CXCL7-containing supernatants from CXCL7 transfected BALB/3T3 in the neutrophil migration assay confirmed that canine CXCL7 had chemoattractive activity for neutrophils. We then used canine monocyte derived DCs to generate CXCL7 for the rest of the experiment. Expression of CXCL7 by DCs treated with LPS, IL-1beta, IL-6, TGF-beta, TNF-alpha, or IFN-gamma was compared using real-time RT-PCR and Western blotting. When treated with IL-1beta, IL-6, TNF-alpha, or TGF-beta, canine DCs expressed significantly higher levels of CXCL7 mRNA and protein than when treated with IFN-gamma or LPS. It is concluded that dog DCs express high levels of the neutrophil chemotactic factor CXCL7 when stimulated by proinflammatory cytokines, including IL-1beta, IL-6, TNF-alpha, or TGF-beta, and may play an important role in modulating inflammatory responses. PMID- 20022387 TI - Peripheral origin of IL-1beta production in the rodent hippocampus under in vivo systemic bacterial lipopolysaccharide (LPS) challenge and its regulation by P2X(7) receptors. AB - In this study we showed that in vivo bacterial lipopolysaccharide (LPS) challenge elevated IL-1beta level in the rodent hippocampus. Antagonists of P2X receptors inhibited LPS-induced IL-1beta level with a pharmacological profile similar to that of P2X(7)R and their inhibitory effect was attenuated in the absence of P2X(7)R. In wild-type mice, LPS overexpressed mRNA encoding P2X(4) and P2X(7) receptors in the hippocampus and caused also a remarkable increase in the levels of IL-1beta in the serum. The hippocampal increase of IL-1beta has substantially alleviated when contamination of circulating blood cells was excluded by transcardial perfusion, indicating the peripheral origin of hippocampal IL-1beta elevation. These results point to the key role of the endogenous activation of peripheral P2X(7)R in the level of IL-1beta in rodent hippocampus under systemic bacterial endotoxin challenge. PMID- 20022388 TI - Supraglottoplasty outcomes in relation to age and comorbid conditions. AB - OBJECTIVE: To determine if age and comorbid conditions effect outcomes in children undergoing supraglottoplasty for severe laryngomalacia. DESIGN: Retrospective study. SETTING: Urban tertiary-care children's hospital. PATIENTS: Children undergoing supraglottoplasty for severe laryngomalacia between February 2004 and July 2008. 56 patients were identified. OUTCOME MEASURES: Persistence of upper airway obstruction, revision surgery (supraglottoplasty), and additional surgery (tracheostomy). RESULTS: 33/56 (58.9%) patients had no comorbid conditions and 23/56 (41.1%) patients had comorbid conditions. In noncomorbid patients, 36.4% of those less than 2 months of age at the time of surgery required revision supraglottoplasty, compared to 5.3% of patients between 2 and 10 months (p<0.05). Compared to the 2-10-month age group, there was a significantly higher percentage of patients with comorbid conditions in the >10 month group (32.1% vs. 79%, p<0.01). Patients with comorbid conditions were diagnosed at a significantly later age than those without (6 mo vs. 2 mo, respectively), and had significantly higher rates of revision supraglottoplasty (47.8% vs. 18.2%) and tracheostomy (39.1% vs. 0.0%). 70% of children with neurological conditions required revision surgery, with 60% requiring tracheostomy. The revision surgery and tracheostomy rates were significantly higher compared to the noncomorbid group (p<0.01 and p<0.0001). Children with cardiac conditions had a higher rate of tracheostomy than noncomorbid children (30% vs. 0%, p<0.01). 16.7% of children with genetic conditions required supraglottoplasty, and none required tracheostomy. CONCLUSIONS: In noncomorbid patients, those undergoing supraglottoplasty less than 2 months of age had a significantly higher rate of revision supraglottoplasty. Patients with neurologic and cardiac comorbidities require tracheostomy at a significantly higher rate than noncomorbid patients. PMID- 20022389 TI - Non-junction functions of pannexin-1 channels. AB - Pannexins are large-pore ion channels with broad expression in the central nervous system (CNS). The channels function by releasing large signaling molecules, such ATP and arachidonic acid derivatives, from neurons and possibly astrocytes. They might also contribute to novel forms of non-synaptic communication in the CNS, thereby affecting synaptic function, astrocytic Ca(2+) wave propagation and possibly regulation of vascular tone in the brain. Panx1 activation in various in vitro pathological conditions implicates these channels in ischemic, excitotoxic and ATP-dependent cell death, whereas Panx coupling with purinergic receptors triggers the inflammasome. Novel functions for the pannexin channels are likely to be discovered as current understanding of how they are regulated in physiological and pathological situations improves. PMID- 20022390 TI - Rehabilitation outcome in home-based versus supervised exercise programs for chronically dizzy patients. AB - We aimed to evaluate the effectiveness of vestibular rehabilitation (VR) exercise between supervised and home-based programs in young and senior age groups of patients with chronic dizziness. Dizziness Handicap Inventory (DHI), Dynamic gait index (DGI), Tinetti fall risk performance scales and Timed "Up and Go" test (TUG) were administered to patients on their initial and follow-up visits for forty-one patients suffering from chronic dizziness. Twenty-eight patients received three 30-min vestibular training exercise sessions per week. Thirteen patients who could not visit our clinic on regular basis were instructed to do the same set of exercises at home, with the same duration and frequency. All scales were evaluated again on their follow-up visits 2 months later. Patients in both groups showed statistically significant improvement in DHI and Tinetti scales. A higher percentage of patients in the supervised-exercise-program (SP) group showed clinically significant outcome improvements. Age was not a predictive factor for rehabilitation outcome. We concluded that for all ages of chronically dizzy patients, 2 months of VR can reduce handicap, improve balance function and a consequent improvement of life quality. Health care professionals need to be educated about the importance of rehabilitation program for treatment of chronic dizziness. PMID- 20022391 TI - Factors affecting attitudes toward care of elderly mothers: urban versus agricultural areas. AB - In our previous study, we examined factors that affect rural people's attitudes toward parental care when their mothers or mothers-in-law become fragile and need 24-h care. Our next task was to examine the factors in an urban area to test external validity. In the previous studies, several factors affecting adult children's attitudes between caring directly for parents or sending parents to a nursing home were indicated. Factors identified included affection, filial obligation, sekentei (i.e., wanting to keep an appearance of taking care), and others. In this study, we examine these factors in a residential urban area, using the same model as before. Results revealed that filial obligation affected attitudes toward care in the case of a mother while affection did in the case of a mother-in-law. This is consistent with the results in the rural setting. Sekentei did not affect attitudes in the urban area, though it had in the rural area. PMID- 20022392 TI - Do red herrings swim in circles? Controlling for the endogeneity of time to death. AB - Studies on the effect of ageing on health care expenditure (HCE) have revealed the importance of controlling for time-to-death (TTD). These studies, however, are subject to possible endogeneity if HCE influences the remaining life expectancy. This paper introduces a 10-year observation period on monthly HCE, socioeconomic characteristics and survivor status to first predict TTD and then use the predicted values as an instrument in the regression for HCE. While exogeneity of TTD has to be rejected, core results concerning the role of TTD rather than age as a determinant of HCE (the 'red herring' hypothesis) are confirmed. PMID- 20022393 TI - Unilateral radiotherapy for tonsil cancer: potential dose distribution optimization with a simple two-field intensity-modulated radiation therapy beam arrangement. AB - BACKGROUND AND PURPOSE: To evaluate the feasibility and dosimetric optimization potential of a unilateral two-field intensity-modulated radiotherapy (IMRT) technique in the curative treatment of lateralized tonsil cancer. MATERIALS AND METHODS: Six patients with lateralized tonsillar carcinoma were treated unilaterally with a two-field IMRT technique (oblique-anterior and oblique posterior fields, with or without collimator and couch rotation). Alternative IMRT plans using seven non-opposed coplanar fields were compared with the two field plans for each patient. RESULTS: Planning target volume (PTV) coverage was excellent with the two-field technique, using a relatively low number of monitor units (MU) (median, 441; range, 309-550). Dose constraints were respected for all organs at risk (OAR). Mean doses to contralateral parotid and submandibular glands were 3.9 and 17.7 Gy, respectively. Seven-field IMRT provided similar PTV coverage, with statistically significant better dose homogeneity and conformality. However, the mean delivered dose to the contralateral parotid (3.9 vs. 9.0 Gy, p=0.001) as well as the mean number of MU (437 vs. 814, p=0.002) and consequently machine time were lower with two-field IMRT. CONCLUSIONS: Unilateral two-field IMRT is a simple and feasible technique providing excellent tumor coverage and optimal OAR sparing while reducing the number of MU and treatment time. PMID- 20022394 TI - Optimizing dose prescription in stereotactic body radiotherapy for lung tumours using Monte Carlo dose calculation. AB - PURPOSE: To define a method of dose prescription employing Monte Carlo (MC) dose calculation in stereotactic body radiotherapy (SBRT) for lung tumours aiming at a dose as low as possible outside of the PTV. METHODS AND MATERIALS: Six typical T1 lung tumours - three small, three large - were constructed centrally, peripherally in the lung, and nearby the thoracic wall, respectively. For each of these, five treatment plans employing dynamic conformal arc technique were made in which the dose was prescribed to encompass the PTV with the prescription isodose level (PIL) set in a range between 50% and 80% of the isocenter dose. Three shells of respectively 10mm thickness around the PTV were constructed to assess the dose in the tissues directly adjacent to the PTV. RESULTS: The PTV was nicely covered (mean 98.8%+/-0.9%) with favourable conformity indices (mean 1.09+/-0.1). Mean doses around the PTVs were 73% (+/-1.3%), 76% (+/-3.5%), and 85% (+/-5.1%) of the prescribed dose in shell 1 for PIL50%, PIL65%, and PIL80%, respectively; 40% (+/-2.6%), 44% (+/-5.1%), 54% (+/-9.3%) in shell 2; and 24% (+/ 1.9%), 26% (+/-3.6%), 33% (+/-6.8%) in shell 3. All normal tissue doses including the integral dose were also consistently worst for PIL80%. Monitor units were 30% higher for PIL65%, and 70% higher for PIL50%, compared with PIL80%. CONCLUSIONS: To improve normal tissue sparing the dose should be prescribed at an isodose lower than 80% of the isocenter dose in SBRT when using conformal arc technique with MC dose calculation. PMID- 20022395 TI - Re-irradiation of metastatic spinal cord compression: a feasibility study by volumetric-modulated arc radiotherapy for in-field recurrence creating a dosimetric hole on the central canal. AB - When local recurrences arise within an irradiated region involving metastatic spinal cord compression, the dose limit to the spinal cord reduces the chance to re-treat the patient by 3D-conformational RT technique. The possibility of using volumetric modulated arc RT by RapidArc was evaluated for dose sparing at spinal cord level and preserving target coverage. A clinically satisfactory PTV coverage and dose sparing to the spinal cord were obtained. An upcoming trial on patients will provide clinical outcomes. PMID- 20022396 TI - Use of a logistic model to assess spoilage by Byssochlamys fulva in clarified apple juice. AB - The percentage P (%) of spoiled bottles (n=40) of clarified apple juice due to Byssochlamys fulva, was modeled by using a logistic model: P = P(max)/1 + exp (k(tau-t)) where P(max) (%) the maximum percentage of spoiled bottles, k (h(-1)) a slope parameter and tau (h) the time for P=P(max)/2. Bottles of pasteurized apple juice were inoculated with B. fulva IOC 4518 ascospores for low and high initial loads, 4.8+/-2.3 ascospores/100mL and 19.3+/-4.6 ascospores/100mL respectively and incubated at 21 degrees C and 30 degrees C. P(max) was not significantly different from 100% except for a low initial load at 21 degrees C. Model parameters were estimated with a good accuracy, RMSE in the range 3.89 7.50. Then the model was used to determine the time for 10% bottles spoiled, t(10%). This time was greater at low initial loads, 57.4 and 104 h at 30 and 21 degrees C respectively, than at high initial loads 23.9 and 75.1h at 30 and 21 degrees C respectively. This study demonstrated that even at a very low initial contamination, clarified apple juice can be easily spoiled by B. fulva highlighting the importance of controlling critical control steps of fruit juice processing (i.e., fruit washing, juice filtration and pasteurization). PMID- 20022397 TI - Fate of Escherichia coli O157:H7 and Salmonella spp. on fresh and frozen cut mangoes and papayas. AB - Escherichia coli O157:H7 and Salmonella infections have been associated with consumption of a number of fruits and vegetables. Although the fate of E. coli O157:H7 and Salmonella on many of these products is well studied, little is known about their behavior on cut mango and papaya. Mangoes and papayas have been associated with four and two documented outbreaks of salmonellosis, respectively. The objective of this study was to evaluate the fate of E. coli O157:H7 and Salmonella on fresh (4 degrees C, 12 degrees C and 23 degrees C) and frozen (-20 degrees C) cut mangoes and papayas. Cut mangoes and papayas were spot inoculated with either a four-strain or five-strain cocktail of E. coli O157:H7 or Salmonella, respectively. Inoculated samples were air dried, placed in containers and stored at 4+/-2, 12+/-2, 23+/-2 and -20+/-2 degrees C. Samples were enumerated following stomaching on nonselective and selective media at days 0, 1, 3, 5, 7, 10, 14, 21 and 28 (4+/-2 and 12+/-2 degrees C); 0, 1, 3, 5 and 7 (23+/-2 degrees C); and 0, 7, 14, 21, 28, 60, 90, 120, 150 and 180 (-20+/-2 degrees C). Population levels (log CFU/g) of fruit were calculated. E. coli O157:H7 and Salmonella have the potential to grow on temperature-abused fresh cut mangoes and papayas held at 23 degrees C. At 12 degrees C, Salmonella populations can grow on cut mangoes and papayas, however E. coli O157:H7 populations only grew on papayas. E. coli O157:H7 and Salmonella survived for 28 days, at 4 degrees C, on refrigerated mangoes and papayas. E. coli O157:H7 and Salmonella can survive on frozen cut mangoes and papayas for at least 180 days. Fresh and frozen cut mangoes and papayas are potential vectors for E. coli O157:H7 and Salmonella transmission. PMID- 20022398 TI - Peripheral endomorphin-1 levels are suppressed in diabetic patients. AB - BACKGROUND: Endomorphins are endogenous ligands selective for mu-opioid receptors, which have been reported to stimulate appetite and regulate glucose homeostasis. But there are no reports about changes in peripheral endomorphin-1 (EM-1) levels in diabetic patients. The aim of this study was to measure plasma EM-1 levels in fasting and postprandial conditions in diabetic patients. METHODS: After an overnight fast, 38 patients (mean age, 67+/-8 years; 17 males and 21 females) and 22 healthy volunteers (mean age, 64+/-9 years; 9 males and 13 females) received a standard breakfast meal with total energy content of 476.1kcal. Blood samples were drawn from each subject in heparinized tubes before breakfast and 2h postprandially. Plasma concentrations of EM-1 were measured by radioimmunoassay (RIA). RESULTS: Comparing with healthy volunteers, EM-1 levels were significantly lower in diabetic patients at both time-points (fasting: 48.38+/-14.13pg/ml vs. 72.71+/-19.62pg/ml, p<0.0001; postprandial: 39.80+/ 12.28pg/ml vs. 62.93+/-21.95pg/ml, p=0.0001). When compared with fasting levels, the postprandial concentrations of EM-1 decreased in both diabetic patients, as well as healthy controls. The absolute value of decrease was not significantly different between the two groups. CONCLUSIONS: Peripheral EM-1 levels were suppressed in diabetic patients and the levels decreased postprandially in both diabetic and healthy volunteers. This implies that EM-1 concentration has correlation with the change in glucose level. Thus, EM-1 could play a role in energy metabolism. PMID- 20022399 TI - Molecular pathology of oxidative stress in diabetic angiopathy: role of mitochondrial and cellular pathways. AB - Diabetes mellitus is characterized by chronic hyperglycaemia and a significant risk of developing micro- and macrovascular complications. Growing evidence suggests that increased oxidative stress, induced by several hyperglycaemia activated pathways, is a key factor in the pathogenesis of endothelial dysfunction and vascular disease. Reactive oxidant molecules, which are produced at a high rate in the diabetic milieu, can cause oxidative damage of many cellular components and activate several pathways linked with inflammation and apoptosis. Among the mechanisms involved in oxidative stress generation, mitochondria and uncoupling proteins are of particular interest and there is growing evidence suggesting their pivotal role in the pathogenesis of diabetic complications. Other important cellular sources of oxidants include nicotinamide adenine dinucleotide phosphate oxidases and uncoupling endothelial nitric oxide synthase. In addition, diabetes is associated with reduced antioxidant defences, which generally contrast the deleterious effect of oxidant species. This concept underlines a potential beneficial role of antioxidant therapy for the prevention and treatment of diabetic vascular disease. However, large scale trials with classical antioxidants have failed to show a significant effect on major cardiovascular events, thus underlying the need of further investigations in order to develop therapies to prevent and/or delay the development of micro- and macrovascular complications. PMID- 20022400 TI - Representation of bone heterogeneity in subject-specific finite element models for knee. AB - Properly representing the heterogeneous distribution of bone tissue material properties is a key step in constructing subject-specific finite element (FE) bone models from computed tomography (CT) data. Conventional methods represent heterogeneity by subjectively grouping bone of similar attenuation together. A new technique characterizing the level of heterogeneity with an objective metric is presented. This technique identifies the minimal level of heterogeneity needed for an accurate FE model. Subject-specific models of the distal femur and proximal tibia were used in this study. An innovative application of an image processing technique in the context of material properties modeling was introduced to facilitate an objective grouping strategy, which gathered together bone based not only on density but also on location thus capturing the natural variation of bone density seen in CT images. A fully heterogeneous model containing unique material properties for each finite element was not necessary to generate an appropriate solution. Von Mises stress, strain energy density, and nodal displacements were predicted within 5% accuracy using a simplified FE femur model containing less than half the number of bone groups of the fully heterogeneous model. Each group contained attenuations varying less than 20% from the group mean. A substantial computational time savings of 60% was gained with the application of the new technique to assign bone mechanical properties. PMID- 20022401 TI - Dynamics of multiple signalling systems: animal communication in a world in flux. AB - The ubiquity of multiple signalling is a long-standing puzzle in the study of animal communication: given the costs of producing and receiving signals, why use more than a single cue? Focusing on sexually selected signals, I argue that dynamic variation in selection pressures can often explain why multiple signals coexist. In contrast to earlier research, which has taken a largely static view of the world, new insights highlight how fluctuations in ecological and social environments, as well as non-equilibrium dynamics intrinsic to coevolutionary systems, can maintain both multiple redundant and non-redundant signals. Future challenges will include identifying the circumstances under which environmental fluctuations lead to multiple signalling, and the consequences of such fluctuations for speciation in multiple-signalling species. PMID- 20022402 TI - Ascorbate regulation of 2-Cys peroxiredoxin-A promoter activity is light dependent. AB - The 2-Cys peroxiredoxin-A (2CPA) promoter is a model promoter to study redox and ABA-dependent stress signaling. Here, an Arabidopsis reporter gene line expressing luciferase under control of the 2CPA promoter was used to study the impact of ascorbate on reporter gene transcription in a series of protoplast and leaf slice incubation experiments. It was shown that ascorbate has a dual function on gene expression regulation. First, a comparison of responses to ascorbate, dehydroascorbate and reduced and oxidized glutathione demonstrated that ascorbate feeding supports gene expression regulation by increasing the catalytic capacity in redox signaling, as defined by the concentration of low molecular weight antioxidants and their oxidized counterparts. Second, ascorbate had a specific and light-dependent effect on 2CPA transcription, which cannot be substituted by reduced glutathione. Based on the differences between ascorbate and glutathione in the subcellular redox-cycling capacities, it is concluded that ascorbate feeding modulates chloroplast-specific regulation of 2CPA expression. PMID- 20022403 TI - Evaluating the impact of a hand hygiene campaign on improving adherence. AB - We monitored compliance with hand hygiene (HH) by direct observation in 3 hospitals in Cantabria, Spain before and after implementation of an HH informational campaign, separately analyzing the effect of a training program. We report that training plus an informational campaign doubled the probability of HH, whereas the informational campaign without training decreased adherence, acting as a deleterious factor in HH adherence. PMID- 20022404 TI - Infection control nurse specialist education in Korea. AB - With the amendment of the Medical Service Act in 2003, the infection control nurse specialist system was launched as one of several specialties for nursing in Korea. Nurse specialists are certified through the national qualifying examination after graduating from a specialist nursing program with core and specialty courses at a graduate school approved by the Ministry of Health and Welfare. This article describes graduate preparation and certification for infection control nurse specialists in Korea. PMID- 20022405 TI - Exposure to hospital roommates as a risk factor for health care-associated infection. AB - BACKGROUND: Numerous patient- and hospital-level characteristics have been established as risk factors for the transmission of health care-associated infections (HAIs). Few studies have quantitatively assessed the impact of exposure to hospital roommates on the acquisition of infections. This study evaluated the association between roommate exposures and the risk of HAIs. METHODS: A retrospective cohort of adult patients admitted to a Canadian teaching hospital between June 30, 2001, and December 31, 2005, was studied. Exposures were characterized as total daily roommate exposures and daily unique roommate exposures. Outcomes examined were methicillin-resistant Staphylococcus aureus (MRSA), vancomycin-resistant Enterococcus (VRE), and Clostridium difficile. RESULTS: The number of roommate exposures per day was significantly associated with MRSA and VRE infection or colonization (MRSA: hazard ratio [HR] = 1.10, 95% confidence interval [CI] = 1.05 to 1.15; VRE: HR = 1.11, 95% CI = 1.02 to 1.21), and with C difficile infection (HR = 1.11, 95% CI = 1.03 to 1.19). A significant association also was found for number of unique roommate exposures per day and VRE (HR = 1.15, 95% CI = 1.02 to 1.28). CONCLUSIONS: The significant associations found between daily roommate exposures and the infection outcomes suggest a possible role for limiting patient-to-patient contact in an infection prevention and control program in this facility. These findings have implications for the deployment and design of acute care hospitals. PMID- 20022406 TI - The pandemic influenza planning process in Ontario acute care hospitals. AB - BACKGROUND: There will be little time to prepare when an influenza pandemic strikes; hospitals need to develop and test pandemic influenza plans beforehand. METHODS: Acute care hospitals in Ontario were surveyed regarding their pandemic influenza preparedness plans. RESULTS: The response rate was 78.5%, and 95 of 121 hospitals participated. Three quarters (76.8%, 73 of 95) of hospitals had pandemic influenza plans. Only 16.4% (12 of 73) of hospitals with plans had tested them. Larger (chi(2) = 6.7, P = .01) and urban hospitals (chi(2) = 5.0, P = .03) were more likely to have tested their plans. 70.4% (50 of 71) Of respondents thought the pandemic influenza planning process was not adequately funded. No respondents were "very satisfied" with the completeness of their hospital's pandemic plan, and only 18.3% were "satisfied." CONCLUSION: Important challenges were identified in pandemic planning: one quarter of hospitals did not have a plan, few plans were tested, key players were not involved, plans were frequently incomplete, funding was inadequate, and small and rural hospitals were especially disadvantaged. If these problems are not addressed, the result may be increased morbidity and mortality when a virulent influenza pandemic hits. PMID- 20022407 TI - Outbreak of late-onset group B Streptococcus in a neonatal intensive care unit. AB - BACKGROUND: In September 2007, the Tennessee Department of Health was notified of a cluster of late-onset group B streptococcal (GBS) infections in a neonatal intensive care unit (NICU). Outbreaks of late-onset GBS are rare. METHODS: A case was defined as culture-confirmed invasive GBS infection in a neonate aged > or =7 days, identified in hospital A during August 23 to September 6, 2007. We reviewed medical records; examined NICU microbiology reports; and performed serotyping, pulsed-field gel electrophoresis (PFGE), and multilocus sequence typing (MLST) on invasive isolates. Maternal GBS screening, prophylaxis, and infection control policies were reviewed and staff practices observed. RESULTS: Five cases of late onset GBS were identified. None of the mothers of the infants received optimal GBS prophylaxis. Patient isolates were of 2 serotypes, 3 PFGE patterns, and 2 MLST patterns. Three isolates were indistinguishable on subtyping. These 3 cases were clustered in time. No common health care providers were identified. Infection control deviations in the NICU were observed. CONCLUSION: We identified a multiclonal cluster of 5 late-onset GBS cases. Multiple factors likely contributed to the outbreak, including nosocomial transmission of GBS. Further efforts to prevent late-onset GBS disease are necessary. PMID- 20022408 TI - Effect of Coenzyme Q10 in mitigating oxidative DNA damage in Down syndrome patients, a double blind randomized controlled trial. AB - Down syndrome (DS) is a chromosomal abnormality (trisomy 21) associated with a complex phenotype. Oxidative stress is known to play a major role in this pathology both due to genetic and epigenetic factors, suggesting that oxidative imbalance contributes to the clinical manifestation of DS. In particular, the implications of oxidative DNA damage in Down syndrome has been linked with neurodegeneration. Here we report the results of a double blind controlled trial aimed at investigating the protective effect of Coenzyme Q(10) on DNA oxidation in this clinical setting using the single cell gel electrophoresis technique. PMID- 20022409 TI - [Nurse prescribing: is it something demanded and really required?]. PMID- 20022410 TI - Direct and indirect effects of cytomegalovirus: can we prevent them? PMID- 20022411 TI - New pyrazoles incorporating pyrazolylpyrazole moiety: synthesis, anti-HCV and antitumor activity. AB - Three series of novel pyrazole derivatives 2b-d, 4a-d and 6a-d were synthesized via two step procedure that utilizes hydrazonoyl chlorides 1a-d and enaminones 3a d and 5a-d, respectively as starting materials. The structures of all the newly synthesized products have been established on the basis of analytical and spectral data. Moreover, some of the products 2-6 were tested against HCV and Subacute Sclerosing Panencephalitis (SSPE). In addition, compounds 2-6 were also tested for the inhibition of peroxynitrite-induced tyrosine nitration and antioxidant activity. The tested compounds are highly effective at very low concentration as anti-HCV, SSPE antioxidant and anticancer in the following ascending order 2d, 4c, 6b, 3b, 6c, 4d, 2b, 2c, 2a, 6a, 5b, 5a, 3a, 4b and 5c. It is worth to mention that all tested compounds are more potent than the reference standards used for comparing activity. All the measurements revealed that the mechanism of action of the anti cancer activities of all the tested compounds is topoisomerase I inhibitor. PMID- 20022412 TI - Synthesis of new chalcone derivatives containing acridinyl moiety with potential antimalarial activity. AB - A series of novel chalcones bearing acridine moiety attached to the amino group in their ring A have been synthesized through noncatalyzed nucleophilic aromatic substitution reaction between various 3'-aminochalcone or 4'-aminochalcones and 9 chloroacridine. The synthesized chalcone derivatives have been characterized and screened for in vitro antimalarial activity against Plasmodium falciparum NF-54. All the chalcones showed complete inhibition at concentration of 10 microg/mL and above while three compounds showed significant inhibition at concentration of 2 microg/mL. The three most active chalcone derivatives were screened for in vivo activity as well, but no significant inhibition in parasitaemia was observed when given intraperitoneally to Plasmodium yoelii infected mice model. PMID- 20022413 TI - [So, doctor, should I take the pandemic flu shot]. PMID- 20022414 TI - Multi-segmental torso coordination during the transition from sitting to standing. AB - BACKGROUND: Research into the multi-segmental mobility of the torso could add to our understanding of the contributions of the head and torso to human movement. The purpose of this study was to determine the motion and temporal coordination of the head and multiple torso segments during the sit-to-stand task. METHODS: Thirty-two young, healthy participants performed five trials of the sit-to-stand movement and 6s of sitting. Range of motion and patterns of peak flexion and extension of six segments and joints and cross correlation of pairs of the six torso segments were analyzed from 3-D kinematic data. FINDINGS: Sagittal range of motion for torso joints during the sit-to-stand task was greater than during sitting trials; motion at the lumbar/pelvis joint was greater than at other torso joints. Peak flexion of torso joints occurred earlier than peak extension. Cross correlations at zero lag and time lags of maximum cross correlations varied such that there was greater temporal coordination of intermediate torso segments compared to pairs including the head and pelvis. There was greater temporal coordination of adjacent segment pairs than for pairs that were less proximal to each other. INTERPRETATION: A high degree of mobility occurs within the torso during the sit-to-stand task. Varying coordination patterns suggest that there are regional differences in movement timing within the torso that may relate to segmental differences in functional roles. Employing multi-segmental torso models may indicate different movement strategies within a healthy population and could highlight differences between clinical populations. PMID- 20022415 TI - mRNA expression of a cadmium-responsive gene is a sensitive biomarker of cadmium exposure in the soil collembolan Folsomia candida. AB - The gene expression of environmental organisms is useful as a biomarker of environmental pollution. One of its advantages is high sensitivity. We identified the cDNA of a novel cadmium-responsive gene in the soil collembolan Folsomia candida. The deduced protein, designated "metallothionein-like motif containing protein" (MTC), was cysteine-rich and contained a metallothionein-like motif with similarity to metallothionein, but had a much longer sequence than metallothionein and contained repeated sequences of amino acids. Expression of MTC mRNA was sensitively induced by cadmium exposure at 0.3 mg/kg of dry food, a concentration at which toxic effects are not observed, but expression was not affected by gamma-ray exposure (an inducer of oxidative stress). These findings suggest that MTC is involved in cadmium-binding processes rather than in oxidative-stress responses. In conclusion, we suggest that gene expression of MTC may be a candidate biomarker for detecting low levels of cadmium contamination in soil. PMID- 20022416 TI - Delivery room resuscitation of near-term infants: role of the laryngeal mask airway. AB - AIM: This observational study aims to describe: (1) the use of positive pressure ventilation (PPV) for resuscitation in the delivery room among newly born near term infants; (2) the methods used for PPV resuscitation [e.g., bag-facial mask (BFM), laryngeal mask airway (LMA), endotracheal tube (ETT)]; and (3) the association of each device with short-term neonatal outcomes. METHODS: We identified near-term (34 0/7-36 6/7 weeks) infants delivered at the Padua University Hospital (Padua, Italy) during the years 2002-2006. The mode of delivery, gestational age, birth weight, Apgar scores, methods of resuscitation and respiratory outcome after NICU admission were analysed. RESULTS: During the 5 year study period, 921 (4.9%) near-term infants were identified from a total of 18,641 live births. PPV was provided in the delivery room to 86 (9.3%) of these infants. Among them, 36 (41.8%) were managed by LMA, 34 (39.5%) by BFM and 16 (18.6%) by ETT. Thirty-four (39.5%) resuscitated near-term infants were admitted to the Neonatal Intensive Care Unit (NICU): 15 (44.1%) after BFM, 12 (75%) after ETT and seven (19.4%) after LMA. Resuscitation with an ETT was associated with an increased rate of respiratory distress syndrome when compared with either BFM or LMA. Resuscitation with an LMA was associated with a lower rate of NICU admission and shorter length of stay when compared with either BFM or ETT. CONCLUSION: The LMA is an effective device for primary airway management of near-term infants and for secondary airway management among near-term infants failing BFM or ETT resuscitation. PMID- 20022417 TI - Critical incidents related to cardiac arrests reported to the Danish Patient Safety Database. AB - BACKGROUND: Critical incident reports can identify areas for improvement in resuscitation practice. The Danish Patient Safety Database is a mandatory reporting system and receives critical incident reports submitted by hospital personnel. The aim of this study is to identify, analyse and categorize critical incidents related to cardiac arrests reported to the Danish Patient Safety Database. METHODS: The search terms "cardiac arrest" and "resuscitation" were used to identify reports in the Danish Patient Safety Database. Identified critical incidents were then classified into categories. RESULTS: One hundred and seven reports describing 122 separate incidents were identified and classified into incidents related to: alerting the resuscitation team (n=32; 26%), human performance (n=22; 18%), equipment failure (n=19; 16%), resuscitation equipment not available (n=13; 11%), physical environment (n=14; 11%), insufficient monitoring (n=14; 11%), and medication error (n=8; 7%). CONCLUSION: Critical incidents related to cardiac arrest occur due to logistical, technical, teamworking and knowledge problems. These findings should be considered when planning education and implementing resuscitation practice. PMID- 20022418 TI - Unusual proximal heme pocket geometry in the deoxygenated Thermobifida fusca: A combined spectroscopic investigation. AB - The spectroscopic properties of the deoxygenated truncated hemoglobin from the actinobacterium Thermobifida fusca have been investigated by means of extended X ray absorption fine structure (EXAFS), X-ray absorption near edge structure (XANES), and near infrared spectroscopies both at room and cryogenic temperatures. At room temperature the near infrared charge transfer band III occurs at 772nm, a value that is unusually high for a canonical deoxygenated hemoglobin species, and can only be found as a transient species after photolysis in vertebrate hemoglobins and myoglobins or under strongly dehydrating conditions. EXAFS and XANES quantitative analyses, carried out in parallel with deoxygenated horse myoglobin, revealed an unusually short iron-histidine distance 1.90+/-0.03A, significantly shorter than the deoxygenated horse myoglobin distance of 2.11+/-0.02A. These findings provide novel structural basis for discussing the fine structural geometry of the proximal site, and eventually mapping the coordinates of the metal with respect to the pyrrole nitrogens and the proximal histidine nitrogen. PMID- 20022419 TI - Security risks in nuclear waste management: Exceptionalism, opaqueness and vulnerability. AB - This paper analyses some potential security risks, concerning terrorism or more mundane forms of crime, such as fraud, in management of nuclear waste using a PEST scan (of political, economic, social and technical issues) and some insights of criminologists on crime prevention. Nuclear waste arises as spent fuel from ongoing energy generation or other nuclear operations, operational contamination or emissions, and decommissioning of obsolescent facilities. In international and EU political contexts, nuclear waste management is a sensitive issue, regulated specifically as part of the nuclear industry as well as in terms of hazardous waste policies. The industry involves state, commercial and mixed public-private bodies. The social and cultural dimensions--risk, uncertainty, and future generations--resonate more deeply here than in any other aspect of waste management. The paper argues that certain tendencies in regulation of the industry, claimed to be justified on security grounds, are decreasing transparency and veracity of reporting, opening up invisible spaces for management frauds, and in doing allowing a culture of impunity in which more serious criminal or terrorist risks could arise. What is needed is analysis of this 'exceptional' industry in terms of the normal cannons of risk assessment - a task that this paper begins. PMID- 20022420 TI - Life cycle cost evaluation of the digital opacity compliance system. AB - The US Environmental Protection Agency (EPA) has established EPA Reference Method 9 (Method 9) as the preferred enforcement approach for verifying compliance with federal visible opacity standards. While Method 9 has an extensive history of successful employment, reliance on human observers to quantify visible emissions is inherently subjective, a characteristic that exposes Method 9 results to claims of inaccuracy, bias and, in some cases, outright fraud. The Digital Opacity Compliance System (DOCS), which employs commercial-off-the-shelf digital photography coupled with simple computer processing, is a new approach for quantifying visible opacity. The DOCS technology has been previously demonstrated to meet and, in many cases, surpass the Method 9 accuracy and reliability standards (McFarland et al., 2006). Beyond its performance relative to Method 9, DOCS provides a permanent visual record of opacity, a vital feature in legal compliance challenges. In recent DOCS field testing, the opacity analysis of two hundred and forty one (241) regulated air emissions from the following industrial processes: 1) industrial scrubbers, 2) emergency generators, 3) asphalt paving, 4) steel production and 5) incineration indicated that Method 9 and DOCS were statistically equivalent at the 99% confidence level. However, a life cycle cost analysis demonstrated that implementation of DOCS could potentially save a facility $15,732 per trained opacity observer compared to utilization of Method 9. PMID- 20022421 TI - The modified Clavien system: a plea for a standardized reporting system for surgical complications. PMID- 20022422 TI - In vitro release of adenosine triphosphate from the urothelium of human bladders with detrusor overactivity, both neurogenic and idiopathic. AB - BACKGROUND: There is increased evidence to suggest a role for nonadrenergic noncholinergic neurotransmission in the pathogenesis of bladder dysfunction. OBJECTIVE: In this set of experiments, we have assessed the contribution of the urothelium to purinergic activity by quantifying the amount of adenosine triphosphate (ATP) released from the urothelium of patients with idiopathic detrusor overactivity (IDO) and with neurogenic detrusor overactivity (NDO) and comparing these releases to those of controls. DESIGN, SETTING, AND PARTICIPANTS: Bladder tissue with urodynamically and clinically proven NDO (n=8) and IDO (n=8) were included in this study. The carefully dissected urothelium was stimulated by mechanically stretching as well as electrically stimulating and the ATP; thus, release was quantified. MEASUREMENTS: We used a Lucy Anthos 1 luminometre (Anthos Labtec Instruments GmBH, Wals, Austria) to perform the assay. The results were analysed using Stingray software (Dazdaq Ltd, Brighton, UK). RESULTS AND LIMITATIONS: Both mechanical stretch and electric field stimulation (EFS) led to increased ATP release in both sets of tissues with overactivity compared to the controls; this rise was even more significant for the IDO urothelium (2416.7+/ 479.8 pmol/g [p<0.005]) than for the NDO urothelium (133.1+/-22.4 pmol/g [p<0.01]); values for the controls were 77.6+/-16.2 pmol/g. ATP release following mechanical stretch was more sensitive to tetrodotoxin in bladders with NDO compared to those with IDO as well as to the controls, with ATP levels falling from 233.5+/-20.7 pmol/g to 107.2+/-11.6 pmol/g, expressed as percentage of basal levels (p<0.002). The experiments were performed in vitro, and the female patients were a mix of peri- and postmenopausal states. CONCLUSIONS: These experiments suggested a significant rise in ATP release from the urothelium of bladders with NDO as well as those with IDO in comparison to controls. Most of the ATP released from bladders with NDO is primarily from neuronal sources. PMID- 20022423 TI - Candida parapsilosis meningitis associated with shunt infection in an adult male. AB - Candida parapsilosis is a very rare cause of meningitis. Though several cases have now been reported in neonates and children, only one has been described in an adult. We report on a 55-year-old male that was admitted due to altered mental status. He had recent sinus drainage and polypectomy, craniotomy with drainage of brain abscess, and ventriculo-peritoneal shunt placement. On admission, imaging studies showed no evidence of shunt dysfunction but did reveal extensive white matter decreased attenuation. Microscopic examination of the first 10 daily cerebrospinal fluid (CSF) cultures revealed yeast. Flucytosine and liposomal amphotericin B were started and externalization of shunt was performed on day 3. On day 8, CSF culture from admission grew C. parapsilosis; fluconazole was added. On day 10, daily CSF still showed yeast and cultures consistently grew C. parapsilosis. Shunt was removed and bilateral ventriculostomy drains were inserted. CSF after procedure as well as at follow-up examinations throughout his 3-month hospitalization were negative for yeast. Extended treatment with flucytosine and fluconazole was initiated. At 8-month follow-up, successful treatment of C. parapsilosis infection without recurrence was confirmed. This case underscores the need for suspicion of C. parapsilosis as a cause of meningitis after invasive surgeries in adults. PMID- 20022424 TI - Optimization of the parameters for decolourization by Aspergillus niger of anaerobically digested distillery spentwash pretreated with polyaluminium chloride. AB - Molasses spentwash from distilleries is characterized by high COD and colour. The fungal decolourization of anaerobically digested molasses spentwash requires significant dilution. In this study, decolourization by Aspergillus niger isolate IITB-V8 was performed on polyaluminium chloride (PAC) treated anaerobically digested spentwash without dilution of wastewater. Optimization of parameters was studied using statistical experimental designs. In the first step, Plackett Burman design was used for screening the important parameters. Glucose was taken as the carbon source for the growth of A. niger. KH(2)PO(4) and pH were found to be the important factors affecting decolourization. In the second step, Box Behnken design was used to determine the optimum level of each of the significant parameters. A second-order polynomial was determined by the multiple regression analysis of the experimental data. The optimum values for the important factors to achieve maximum decolourization of 68.4% were 5.5 g/L Glucose, 1.2 g/L KH(2)PO(4) and 5 pH. The determination coefficient (R(2)) was 0.9973, which ensures adequate credibility of the model. The total decolourization obtained after fungal treatment was 86.8% which indicates fungal decolourization after pretreatment with PAC is a viable option for the treatment of digested molasses spentwash. PMID- 20022425 TI - Fast and efficient mesoporous adsorbents for the separation of toxic compounds from aqueous media. AB - The effect of cationic template on the adsorption of chromium (VI), furfural and copperphthalocyanine-3,4',4'',4'''-tetrasulfonic acid tetrasodium salt (CuPc) in MCM-41 and MCM-48 mesoporous materials was investigated in this work. We used cetyltrimethylammonium bromide (CTAB) as the cationic template and sodiummetasilicate (for MCM-41) and tetraethyl-orthosilicon (for MCM-48) as the silica source for the synthesis of mesoporous materials. The properties of synthesized samples were characterized with XRD-low angle, SEM, N(2) adsorption desorption and TG-DT analysis. The extent of adsorption was investigated as a function of solution pH, shaking speed, contact time, analyte concentration, reaction temperature and supporting electrolyte (sodium chloride) concentration. Langmuir and Freundlich isotherms were used to model the adsorption equilibrium data. The as-synthesized mesoporous samples showed very high adsorption capacity for the analytes and adsorption uptakes were rapid on the adsorbents reaching equilibrium in less than 2 h. The materials without surfactant did not show significant affinity for the analytes. PMID- 20022426 TI - Seawater ecotoxicity of monoethanolamine, diethanolamine and triethanolamine. AB - Monoethanolamine (MEA), diethanolamine (DEA) and triethanolamine (TEA) are compounds with potential acute, sub-chronic and chronic toxicity effects towards aquatic species. A literature review highlighted the existence of a gap in the knowledge on their toxicity with saltwater testing species. A battery of toxicity tests including the alga Phaeodactylum tricornutum Bohlin, the bivalve molluscs Crassostrea gigas (Thunberg) and Mytilus galloprovincialis (Lmk), and the crustacean Artemia franciscana, was considered to update and improve the existing ecotoxicological information. Data were provided as the Effective Concentration that induces a 50% effect in the observed population (EC50), Lowest Observed Effect Concentration (LOEC) and No Observed Effect Concentration (NOEC). EC50, LOEC and NOEC values were compared with a reviewed database containing the existing ecotoxicological data from saltwater organisms. PMID- 20022427 TI - An anti-nociceptive role for ceftriaxone in chronic neuropathic pain in rats. AB - Glial glutamate transporter-1 (GLT-1) plays an essential role in the maintenance of glutamate homeostasis and is involved in the development and maintenance of pathological pain. The present study was undertaken (1) to observe the anti nociceptive effects of ceftriaxone (Cef) in a chronic neuropathic pain model induced by chronic constrictive nerve injury (CCI) of the sciatic nerve and (2) to identify the role of spinal GLT-1 in the process. CCI induced significant thermal hyperalgesia and mechanical allodynia, which began from postoperative day 3 and lasted to day 21. This long-term hyperalgesia was accompanied by significant down-regulation of GLT-1 expression in the L4-L6 segments of the spinal dorsal horn, as revealed by immunohistochemistry and Western blot. Intraperitoneal preventive and therapeutic administration of Cef effectively prevented or reversed, respectively, the development of thermal hyperalgesia, mechanical allodynia, and GLT-1 down-regulation in the spinal dorsal horn. To further determine whether the above anti-nociceptive effects of Cef are a result of the up-regulation of spinal GLT-1 expression and its function, we further observed the effects of intrathecal administration of Cef in the same model. It was found that intrathecal administration of Cef led to the specific up regulation of GLT-1 expression and glutamate uptake ((3)H-glutamate) in the spinal dorsal horn, and similar anti-nociceptive effects to those of intraperitoneal administration of Cef. The above effects of intrathecal Cef administration were all significantly inhibited by intrathecal administration of GLT-1 antisense oligodeoxynucleotides (As-ODNs). These results indicate that Cef plays an anti-nociceptive role by up-regulating spinal GLT-1 expression and its function. PMID- 20022428 TI - Reliability of intraoral quantitative sensory testing (QST). AB - The German Research Network on Neuropathic Pain (DFNS) has recommended a protocol with 13 quantitative sensory testing (QST) measures for detecting somatosensory abnormalities. Reliability is an important scientific property and has been adequately tested for cutaneous QST. This study evaluates intraoral sites for which no reliability trials have yet been published. Inter- and intra-examiner reliability of 13 QST measures at intra- and extraoral trigeminal sites were investigated. Twenty-one healthy volunteers from Malmo University, Malmo, Sweden (13 women and 8 men, mean age 40.4 years, range 24-71) participated. Two independent examiners previously trained in the DFNS QST protocol examined the participants using the entire protocol. Each participant was examined twice on the same day, once by each examiner (inter-examiner reliability). After 1-3 weeks, one examiner re-examined all participants (intra-examiner reliability). The measurements were made on the skin of the right cheek, the tip of the tongue, and bilaterally on the gingival mucosa of the upper premolar region. The intraclass correlation coefficient (ICC) or kappa was used to calculate variations. Most tests had acceptable to excellent inter-examiner (ICC 0.41-0.89) and intra-examiner (ICC 0.43-0.87) reliability. For each test, inter- and intra examiner reliabilities at intra- and extraoral sites were similar. No significant differences between right and left sides were found intraorally. We conclude that inter- and intra-examiner reliabilities of most QST measures are acceptable for assessing somatosensory function in the orofacial region. PMID- 20022429 TI - Fleas and ticks as vectors of Leishmania spp. to dogs: caution is needed. PMID- 20022430 TI - Pulsed dye laser in burn scars: current concepts and future directions. AB - Hypertrophic scarring after partial-thickness burns is common, resulting in raised, erythematous, pruritic, and contracted scars. Treatment of hypertrophic scars, especially on the face, is challenging and has high failure rates. Excisional treatment has morbidity and can create iatrogenic deformities. After an extensive experience over 10 years with laser therapy for the treatment of difficult scars, the pulsed dye laser (PDL) has emerged as a successful alternative to excision in patients with hypertrophic burn scars. Multiple studies have shown its ability to decrease scar erythema and thickness while significantly decreasing pruritus and improving the cosmetic appearance of the scar. The history of laser therapy and the mechanism of action and results of the PDL in burn scars will be reviewed. The PDL should become an integral part of the management of burn scarring and will significantly decrease the need for excisional surgery. PMID- 20022431 TI - Feasibility and potential effect of a low-cost virtual reality system on reducing pain and anxiety in adult burn injury patients during physiotherapy in a developing country. AB - OBJECTIVE: The purpose of this study was to ascertain the feasibility and potential effect of a low-cost VR system (eMagin Z800 3DVisor), used in conjunction with pharmacological analgesia, on reducing pain and anxiety in adult burn patients undergoing physiotherapy treatment, compared to pharmacologic analgesia alone at a South African hospital. STUDY DESIGN: Single-blind, within subject study design. METHODS: Pain and anxiety outcome measures were measured by a blinded assessor using the Numeric Pain Rating Scale and Burn Specific Pain and Anxiety Scale. Box-and-whisker plot method, Chi-square tests as well as the Student's paired t-test were used to analyze data. MAIN FINDINGS: Eleven eligible adult burn patients consented to participate in this study (3 female, 8 male; median age 33 years: range 23-54 years). A marginal (p=0.06) to insignificant (p=0.13) difference between the two sessions (analgesia with VR and analgesia without VR) in reducing pain was found. No significant difference (p=0.58) was found between the two sessions (analgesia with VR and analgesia without VR) for anxiety. CONCLUSION: There seems to be a trend that the low-cost VR system, when added to routine pharmacological analgesics, is a safe technique and could be of considerable benefit if implemented into the pain management regime of burn units at a South African hospital. PMID- 20022432 TI - Transfusion of sickle cells may be a therapeutic option for patients suffering metastatic disease. AB - Red blood cells from patients with sickle cell disease will sickle under conditions of hypoxemia and acidosis which is a similar milieu found in malignant tumors. While control of tumor angiogenesis has long been a goal of cancer therapy, selective occlusion of tumor blood supply may be achieved by transfusion of sickle cells into patients who suffer metastatic cancer. Although this potential therapy has not been previously reported in the medical literature, the concept may have been elusive to medical mainstream thinking because it requires transfusion of diseased cells. For this therapy to be effective, other environmental factors may need to be manipulated such inducing mild hypoxemia or hypercarbia (respiratory acidosis) to induce red cell sickling. Preliminary evidence supportive of this therapeutic approach to cancer treatment is provided by case evidence that sickle cell occlusion of a malignant brain tumor (glioma) produced tumor necrosis. Also sickle cells have been successfully transfused into primates. Furthermore, donor blood is crossmatched and transfused into patients suffering from sickle cell disease regularly in clinics and this procedure is associated with acceptable morbidity. Most importantly, animal models of sickle cell disease and cancer currently exist, and this theory could be tried with available technologies including ultrasound detection of vaso-occlusion. While the proposed therapy may not cure metastatic cancer, this treatment could prove useful for decreasing the size and perhaps the pain from metastatic tumor burden. Therefore, it is hypothesized that ABO Rh compatible crossmatched sickle cells transfused into patients who suffer metastatic cancer under controlled conditions of blood oxygenation and pH will selectively produce vaso-occlusive infarcts in malignant tumors and be a useful therapy. The author hopes for further investigations. PMID- 20022433 TI - House-dust mites in our homes are a contamination from outdoor sources. AB - Avoidance advices for house-dust mite sensitized persons are currently based upon the idea, that the mites (Dermatophagoides spp.) are part of the indoor fauna. A closer look at development stages in the house-dust samples shows, however, that only the mites' active stages are present there and that the stages between them, the inactive moulting stages, are absent. Therefore the mites probably do not carry out their life cycles in our dwellings, but are more likely contaminations from the open. Findings of low level concentrations can be explained by mites coming from outdoors and sedimented in accordance with known physical laws. The occasional finding of higher concentrations is the result of synchronized populations of the mites developing outdoors and being passively transported into our homes by wind and dust. The hypothesis explains why we find mites in our homes but nonetheless have no effect of avoidance measures. The verification of the entire hypothesis or part of it may have great impact on the management of the disease house-dust mite allergy. PMID- 20022434 TI - Glutamatergic modulatory therapy for Tourette syndrome. AB - Tourette syndrome (TS) is a neuropsychiatric disorder characterized by the presence of chronic, fluctuating motor and vocal (phonic) tics. The disorder is commonly associated with a variety of comorbidities including obsessive compulsive disorder (OCD), attention deficit hyperactivity disorder (ADHD), school problems, anxiety, and depression. Therapeutically, if tics are causing psychosocial or physical problems, symptomatic medications are often prescribed, typically alpha-adrenergic agonists or dopamine antagonists. Recognizing that therapy is often ineffective and frequently associated with unacceptable side effects, there is an ongoing effort to identify new tic-suppressing therapies. Several lines of evidence are presented that support the use of glutamate modulators in TS including glutamate's major role in cortico-striatal-thalamo cortical circuits (CSTC), the recognized extensive interaction between glutamate and dopamine systems, results of familial genetic studies, and data from neurochemical analyses of postmortem brain samples. Since insufficient data is available to determine whether TS is definitively associated with a hyper- or hypo-glutamatergic state, potential treatment options using either glutamate antagonists or agonists are reviewed. Data from studies using these agents in the treatment of OCD are presented. If validated, modulation of the glutamate system could provide a valuable new pharmacological approach in the treatment of tics associated with Tourette syndrome. PMID- 20022435 TI - The origin of serous ovarian cancer may be found in the uterus: a novel hypothesis. AB - Since 1971 the incessant ovulation theory by Fathalla is widely accepted as theory for ovarian carcinogenesis, supported mainly by epidemiological findings. However, this theory cannot explain the protective effect of hysterectomy and tubal ligation on the incidence of ovarian cancer. Furthermore, never a precursor lesion has been identified in the ovary itself. Although recently the fallopian tube has been proposed as possible site of origin, there are reasons to believe that a precursor lesion for ovarian and pelvic serous carcinoma is located within the uterus. Uterine serous papillary carcinoma (UPSC) resembles serous ovarian and pelvic carcinoma in behavior and prognosis. Its precursor lesion endometrial intraepithelial carcinoma (EIC) is non-invasive and often multifocal in origin. Importantly, these premalignant cells have a loosely cohesive nature and are able to spread to intraperitoneal surfaces easily, thereby often found on the surface of ovaries or in the fallopian tube. We hypothesize that EIC is a precursor lesion of serous ovarian carcinoma, originating in the uterus and spreading into the intraperitoneal cavity via a mechanism as is accepted for endometriosis. To illustrate this, some cases of serous ovarian carcinoma with concordant EIC in the endometrium as only precursor lesions are presented. A paradigm shift with respect to the origin of ovarian cancer from the ovary to the endometrium could have enormous consequences for primary and secondary preventive strategies to decrease the mortality from this disease. PMID- 20022436 TI - Ovarian follicular dynamics and hormonal secretory profiles in guanacos (Lama guanicoe). AB - The objective of the present study was to describe ovarian activity in 11 adult non-mated guanacos, evaluated every second day for 40 days by transrectal ultrasonography and by plasma estradiol-17beta and progesterone concentrations. An inverse relationship (r = -0.29, P < 0.001) was observed between the diameter of the largest ovarian follicle and the total number of follicles indicating that follicular growth in guanacos occurs in waves. The mean duration of follicular wave was 15.1 +/- 4.2 days with a range from 9 to 26. The follicular growth phase was 7.0 +/- 2.4 days, the static phase 3.0 +/- 1.2 days, the regression phase 5.2 +/- 2.1 days and the inter-wave interval was 12.6 +/- 5.6 days. The maximum follicular diameter in each follicular wave was 10.2 +/- 2.1 mm with a range from 7.2 to 16.1 mm. Inter-wave intervals of longer duration were associated with a larger maximum follicle diameter (P < 0.001). Follicular activity alternated between ovaries in 93% of the waves with 48% of dominant follicles occurring in the left and 52% in the right ovary without differences (P > 0.05). Plasma estradiol-17beta concentrations showed a wave-like pattern, varying from 20.0 to 92.1 pmol/L. Plasma progesterone concentrations remained below 1 nmol/L without any ultrasonic evidence of ovulation during the study. These results in guanacos suggest a follicular wave pattern more similar to the llama (Lama glama) than previously described in other South American and Old World camelid species. PMID- 20022437 TI - The application of computerized tomography (CT) to the dental ageing of children and adolescents. AB - INTRODUCTION: Following a mass disaster, the aim of the Disaster Victim Identification process is to establish the identity of the victims. The ageing screening process on victims in Victoria may now be complemented with the use of computerized tomography (CT), where previously any dental ageing analysis was performed using conventional radiographs. The aim of this study was to assess the accuracy of age estimation using the dental ageing method proposed by Moorrees, Fanning and Hunt (MFH) using CT images. Intra- and inter-rater variability between two raters, one experienced and one inexperienced, was also assessed. MATERIALS AND METHODS: The two raters were blinded to the ages of 96 deceased Australian children aged up to 15 years. Using three-dimensional (3D) shaded surface displays (SSD) and reformatted CT images, the age was first estimated based on prior experience alone, followed at a later date by the age estimation utilizing the MFH method. These estimates were then compared to the known chronological age. The results were statistically analyzed in a one-sample t test, using the mean log-ratio of the estimated age to the chronological age. RESULTS: Our findings show that the experienced rater was more accurate in age estimation than the less experienced when using prior experience (p<0.0001). The use of reformatted CT images to perform an ageing estimate using the MFH method was found to systematically underestimate the chronological age by 10% by both raters (p=0.784). There was no significant difference between the two raters. Intra-rater reliability was high (p=0.135). CONCLUSIONS: CT can provide accurate estimates of dental ages. Prior experience with dental ageing and/or CT improves the accuracy. However, with the use of validated ageing charts, inexperienced raters can also achieve accurate age estimates using CT images. PMID- 20022438 TI - Differentiation of methylenedioxybenzylpiperazines (MDBP) by GC-IRD and GC-MS. AB - The substituted benzylpiperazine, 3,4-methylenedioxybenzylpiperazine (3,4-MDBP) and its regioisomer 2,3-methylenedioxybenzylpiperazine (2,3-MDBP) have almost identical mass spectra. Perfluoroacylation of the secondary amine nitrogen of these regioisomeric piperazines gave mass spectra with differences in relative abundance of some fragment ions. However the spectra did not yield any unique fragments for specific identification of one regioisomer to the exclusion of the other compound. Gas chromatographic separation coupled with infrared detection (GC-IRD) provides direct confirmatory data for structural differentiation between the two regioisomers. The mass spectrum in combination with the vapor-phase infrared spectrum provides for specific confirmation of each of the regioisomeric piperazines. The underivatized and perfluoroacyl derivative forms of the ring substituted benzylpiperazines were resolved on a 30-m capillary column containing an Rxi-50 stationary phase. PMID- 20022439 TI - Mild phenotype in Pelizaeus-Merzbacher disease caused by a PLP1-specific mutation. AB - We present the case of a 26 year-old man who developed normally until he began having difficulty walking at age 12. He subsequently became unable to stand at 15 years old and exhibited mental regression and generalized tonic convulsions by age 20. Magnetic resonance imaging revealed incomplete myelination of cerebral white matter, which resembled that of Pelizaeus-Merzbacher disease. By sequencing the proteolipid protein 1 (PLP1) gene, we found a novel mutation (c.352_353delAG (p.Gly130fs)) in the latter half of exon 3 (exon 3B) that is spliced out in the DM20 isoform. Exon 3B mutations are known to cause a mild phenotype since they do not disturb DM20 production. Mutations that truncate PLP1 correlate with a mild phenotype by activating the nonsense-mediated decay mechanism that specifically detects and degrades mRNAs containing a premature termination codon. This attenuates the production of toxic mutant PLP1. The very mild presentation in the present case seems to be derived from the unique nature of the mutation, which preserves DM20 production and decreases mutant PLP1. PMID- 20022440 TI - Novel approaches using alkaline or acid/guanidine treatment to eliminate therapeutic antibody interference in the measurement of total target ligand. AB - Measurement of the total target ligand can help to provide pharmacokinetic (PK) and pharmacodynamic (PD) informations. However, the presence of monocloncal antibody therapeutics (ThAs) interferes with ELISA determinations of the total target proteins. The interferences can cause over- or under-estimation of the target protein analysis. The nature of interferences was dependent upon the ThA, target protein, antibody reagents and assay conditions of the ELISA. We have developed novel alkaline and acid/guanidine treatment approaches to dissociate the protein binding and preferentially denature the ThA. The neutralized target proteins can be determined by ELISA. These methods provide reproducible measurements of total target protein without ThA interference. Serum samples, standards and QCs containing target protein and ThA were treated with alkaline buffer (pH>13) containing casein or acid/guanidine buffer (pH<1). Total target proteins for two different ThA systems were successfully measured and interferences were completely eliminated by the treatments. These methods were successfully applied to analysis in pre-clinical serum samples. PMID- 20022441 TI - New solid modifications of nateglinide. AB - New modifications of the antidiabetic drug nateglinide were found and characterized by means of thermal analysis, vibrational spectroscopy and X-ray powder diffractometry. In particular it has been verified that the product obtained during the final steps of the nateglinide synthesis is the hemihydrate form which melts at about 86 degrees C provided that the adopted experimental conditions hinder the removal of the crystallization water. Otherwise, if the crystallization water is removed, the hemihydrate transforms to a new anhydrous polymorph that melts at 102.8 degrees C. The anhydrous polymorph, if stored at room temperature and humidity, gradually changes to H polymorph while, if stored in water vapour saturated atmosphere, it gets back water and reverts to the hemihydrate form. On the contrary, both an isothermal treatment at 80 degrees C and melt cooling bring to the B polymorph. PMID- 20022442 TI - Recent advances in trace analysis of pharmaceutical genotoxic impurities. AB - Genotoxic impurities (GTIs) in pharmaceuticals at trace levels are of increasing concerns to both pharmaceutical industries and regulatory agencies due to their potentials for human carcinogenesis. Determination of these impurities at ppm levels requires highly sensitive analytical methodologies, which poses tremendous challenges on analytical communities in pharmaceutical R&D. Practical guidance with respect to the analytical determination of diverse classes of GTIs is currently lacking in the literature. This article provides an industrial perspective with regard to the analysis of various structural classes of GTIs that are commonly encountered during chemical development. The recent literatures will be reviewed, and several practical approaches for enhancing analyte detectability developed in recent years will be highlighted. As such, this article is organized into the following main sections: (1) trace analysis toolbox including sample introduction, separation, and detection techniques, as well as several 'general' approaches for enhancing detectability; (2) method development: chemical structure and property-based approaches; (3) method validation considerations; and (4) testing and control strategies in process chemistry. The general approaches for enhancing detection sensitivity to be discussed include chemical derivatization, 'matrix deactivation', and 'coordination ion spray-mass spectrometry'. Leveraging the use of these general approaches in method development greatly facilitates the analysis of poorly detectable or unstable/reactive GTIs. It is the authors' intent to provide a contemporary perspective on method development and validation that can guide analytical scientists in the pharmaceutical industries. PMID- 20022443 TI - Analysis of rare flavonoid C-glycosides in Celtis australis L. by micellar electrokinetic chromatography. AB - This manuscript reports on the first analytical procedure for the determination of flavonoids in Celtis australis. The capillary electrophoretic separation of 8 compounds, most of them flavone C-glycosides, was possible using a borax buffer with pH 9.0, which contained 25mM SDS as detergent and 7.5% of n-butanol as organic modifier. Method validation revealed that the developed assay is repeatable (sigma(rel)or=0.9996) within the tested concentration range. The quantitative analysis of several C. australis samples showed that isovitexin is the most abundant representative (0.06-0.09%), at a rather uniform content of total flavonoids of approx. 0.3% in all specimens. PMID- 20022444 TI - Endometrial and conceptus expression of HoxA10, transforming growth factor beta1, leukemia inhibitory factor, and prostaglandin H synthase-2 in early pregnant pigs with gonadotropin-induced estrus. AB - This study was conducted to evaluate the effect of estrus induction with gonadotropins on endometrial and conceptus expression of HoxA10, transforming growth factor (TGF) beta1, leukemia inhibitory factor (LIF), and prostaglandin H synthase-2 (PGHS-2) during early pregnancy in pigs. Twenty-four prepubertal gilts received 750 IU of pregnant mare serum gonadotropin (PMSG) and 500 IU of human chorionic gonadotropin (hCG) 72h later. Gilts in the control group (n=23) were observed daily for estrus behavior. Endometrial tissue samples, conceptuses, blood serum, and uterine luminal flushings (ULFs) were collected on days 10, 11, 12, and 15 after insemination. There was no effect of estrus induction on estradiol content in ULFs, or on ovulation and fertilization rates in studied gilts. However, the content of progesterone in the blood serum was greater in naturally ovulated gilts in comparison to gonadotropin-treated animals on day 12 of pregnancy (P<0.05). HoxA10 expression was up-regulated in the endometrium of pregnant gilts, with natural ovulation on days 12 (P<0.05) and 15 (P<0.001) in comparison to days 10 and 11. When compared to control gilts, administration of PMSG/hCG resulted in decreased expression of endometrial HoxA10, TGFbeta, LIF, and PGHS-2 on day 12 of pregnancy (P<0.05). Conceptus expression of studied factors was not affected by gonadotropin treatment. Overall, these results suggest improper endometrial preparation for implantation in prepubertal gilts induced to ovulate with PMSG/hCG. PMID- 20022445 TI - The role of retinal and extra-retinal photostimulation in reproductive activity in broiler breeder hens. AB - Photostimulation of retinal photoreceptors, which are sensitive to green light, appears to inhibit reproductive activity in birds, whereas photostimulation of extra-retinal photoreceptors, which are sensitive to red light, accelerates it. The objective of this study was to determine the effect of either retinal or extra-retinal photostimulation on reproductive activities of broiler breeder hens. At 23 wk of age, Cobb hens (N=135) were divided into 9 rooms with individual cages (n=15). At 24 wk of age, 3 rooms were photostimulated (14L:10D) with white light (Control, n=45). Six rooms had 2 parallel lighting systems, red (660 nm) and green (560 nm), which were both on during 6 out of 14 h of the light period. Then, in 3 of these rooms, the green light was turned off and hens were exposed to a total of 14 h of red light (Red, n=45), and in the other 3, the red light was turned off and green lighting continued for a total of 14 h (Green, n=45). The Green group had reduced egg production; reduced plasma concentrations of ovarian steroids; reduced luteinizing hormone (LH)-beta, vasoactive intestinal peptide (VIP), and prolactin mRNA expression; and greater retinal green opsin mRNA expression (P < or = 0.05). The Red group had greater egg production; greater gonadotropin-releasing hormone-I (GnRH-I) and red opsin gene expression in the hypothalamus; and lesser green opsin gene expression in the retina (P < or = 0.05). We suggest that selective photostimulation of extra-retinal photostimulation as opposed to retinal photostimulation is a key factor in the determination of successful reproduction of broiler breeder hens. PMID- 20022446 TI - Expression of Ki-67, PCNA, and p27kip1 in canine pituitary corticotroph adenomas. AB - Pituitary-dependent hypercortisolism (PDH), which is caused by adrenocorticotropic hormone (ACTH)-secreting pituitary adenomas, is a common endocrinopathy in dogs. Dogs with non-enlarged pituitaries harboring a microadenoma have a better prognosis than those with enlarged pituitaries. The aim of this study was to investigate the expression of the proliferation markers Ki-67 and proliferating cell nuclear antigen (PCNA) and the cell-cycle inhibitor p27kip1 in corticotroph adenomas in enlarged and non-enlarged pituitaries. The expression of Ki-67, PCNA, and p27kip1 was analyzed by immunohistochemical staining of 17 pituitary adenoma samples harvested during pituitary surgery in dogs with PDH. The labeling index was calculated by counting the number of immunopositive cells per 1,000 cells. The mean (+/- standard deviation) labeling index for Ki-67 was 8.4%+/-14.2% for the group with enlarged pituitaries, and 8.8%+/-5.5% for the group with non-enlarged pituitaries; that for PCNA was 35.5%+/-12.2% and 37.0%+/-15.5%; and that for p27kip1 was 29.3%+/-22.6% and 42.5%+/-27.9%, respectively. No significant differences in Ki-67, PCNA, and p27kip1 labeling indices were found between enlarged and non-enlarged pituitaries. However, a trend toward significance was observed when comparing the expression of p27kip1 in enlarged pituitaries versus normal pituitary tissue. It is concluded that Ki-67 and PCNA are not useful as proliferative markers for studying the pathobiology of pituitary corticotroph adenomas in dogs. PMID- 20022447 TI - A spline polynomial model to describe serum IGF-I concentration from birth to slaughter in calves: effects of weaning age, pre-weaning concentrate feeding and breed. AB - The statistical analysis of hormones sampled throughout the production cycle is complicated because factors such as age and weight at the measuring date interfere. Spline curves constructed from pieces of low-degree, random-effects polynomials could be used for a more accurate analysis of data. Concentration of insulin-like growth factor-1 (IGF-1), weight gain, and concentrate intake of Parda de Montana (PM) (n=27) and Pirenaica calves (n=14) were modeled with a spline model according to age at weaning, pre-weaning concentrate feeding, and breed. At birth, calves were randomly assigned to early weaning (EW) at 90 d or traditional weaning (TW) at 150 d. During lactation, half of PM calves received concentrates (S), whereas the remainder received no concentrates (NS). After weaning, calves received concentrates on an ad libitum basis until they reached a weight of 450 kg. The spline model had better likelihood than a polynomial of 6 degrees or a split-plot model. Serum IGF-1 concentration was greatly affected by age at weaning and pre-weaning concentrate feeding, but not by breed. In NS calves, IGF-1 concentration was greater in EW than in TW calves from 120 to 300 d, irrespective of breed. During lactation, S calves had greater IGF-1 concentration than NS calves. After weaning, EWNS calves reached the IGF-1 concentration of EWS calves after 4 mo on concentrates, whereas TWNS calves attained IGF-1 concentration of TWS calves after only 2 mo, because of their increased concentrate intake relative to TWS calves. Concentration of IGF-1 was positively correlated with the immediate weight gains and intake, but it was not a good predictor of performance in the long term. PMID- 20022448 TI - Three-dimensional surface deviation maps for analysis of retrieved polyethylene acetabular liners using micro-computed tomography. AB - Micro-computed tomography (micro-CT) has previously been validated for measuring wear volume in polyethylene acetabular liners. The creation of 3-dimensional (3D) articular and backside surface deviation maps using micro-CT is described. An acetabular liner was retrieved after 16.7 years of implantation and scanned with micro-CT along with a new, never-implanted liner of the same size and type. The liner surface geometries were reconstructed and co-aligned. A 3D comparison and cross-sectional analysis was performed. Maximum 3D deviation of the articular surface was -2.48 +/- 0.02 mm, with maximum backside deviation of 0.46 +/- 0.02 mm. Micro-CT can measure surface deviation and therefore calculate the volume of wear plus creep of retrieved acetabular liners, and may be applicable for wear simulator studies and analyzing other polyethylene components including tibial inserts. PMID- 20022449 TI - Guidelines for implant placement to minimize impingement during activities of daily living after total hip arthroplasty. AB - Impingement, both prosthetic and bony, precedes the vast majority of dislocations after total hip arthroplasty and may adversely impact component wear. Reconstructed computer hip models of 8 subjects were used to evaluate hip range of motion for activities of daily living (ADLs) associated with posterior instability and anterior instability. Variables examined included acetabular position, femoral offset, and head size. The majority of flexion ADLs (associated with posterior instability) encountered prosthetic impingement, whereas extension ADLs demonstrated bony impingement with the 45/20 cup placement position. Cup placement in natural anteversion and adduction allowed normal joint motion in anterior and posterior impinging activities. Insufficient femoral offset and smaller head size negatively impacted range of motion. Any anterior cup and posterior cup protrusions greater than 5 mm should be avoided. PMID- 20022450 TI - Gait analysis of knee arthritis treated with hyaluronic acid. AB - Self-reported studies have demonstrated efficacy of hyaluronic acid (HA) therapy. Gait analysis may objectively demonstrate changes associated with HA therapy. Fifty-three consecutive patients with unilateral osteoarthritis (OA) of the knee were evaluated with a validated and sensitive gait laboratory previously used for gonarthrosis. Two 100-meter walks were performed before and after HA therapy. Nineteen parameters were measured. In addition, the subjective response to treatment was correlated with changes in gait parameters. Single-limb support time, velocity, fatigability, and swing phase were not improved. Only double-limb support time was found to be significantly different (P = .04). The remaining gait variables were not changed. Separately, the subset of patients that achieved greater than 70% subjective pain relief had no objective improvements in gait parameters. Hyaluronic acid therapy may result in a placebo effect for the treatment of gonarthrosis. PMID- 20022451 TI - Extensive osteolysis caused by polyethylene particle migration in an anatomical hydroxyapatite-coated hip prosthesis: 10 years' follow-up. AB - We report our 10-year follow-up results of 630 consecutive Anatomique Benoist Giraud I hip prostheses implanted between June 1990 and December 1995. At this time, 520 were satisfactory and 25 had been revised. Although the majority of patients remained asymptomatic at the end of follow-up, the real survivorship of the implant was less than 91% (33 patients who were in the waiting list for revision due to osteolysis at that time were revised by December 2007). On the other hand, radiographic outcomes were of concern: around 90% of patients showed progressive stress shielding and large granulomatous lesions in the proximal femur, and more than 82% of patients exhibited polyethylene wear in excess of 1 mm (mean=1.69 mm). PMID- 20022452 TI - Porous-coated cementless acetabular components without bulk bone graft in revision surgery. AB - We previously reported the average 9.3-year (range, 5-13 years) results of 74 patients (83 hips) with porous-coated acetabular components that were placed without bulk bone graft at revision surgery. Since the previous report, 7 patients (7 hips) died before the minimum follow-up of 10 years, and 1 patient (1 hip) was lost to follow-up. We now report the average 15.6-year (range, 10-20 years) results for 66 patients (75 hips). Three additional acetabular components were removed or revised again: 2 for infection and 1 for dislodgement of the polyethylene liner from the metal shell. Overall, 7 (7%) components required removal or repeat revision. No shell was revised for aseptic loosening, and none was categorized as loose during the entire follow-up period. PMID- 20022453 TI - Metal-on-metal cups cemented into reinforcement rings: a possible new acetabular reconstruction procedure for young and active patients. AB - The purpose of this study was to evaluate the clinical and radiological results of Metasul cups cemented into reinforcement rings for young and active patients. Twenty-three total hip arthroplasties with Metasul cups were cemented into Muller reinforcement rings. Mean follow-up was 6.1 years (5-10). At final follow-up, the Harris hip score increased from 62.2 (39-85) to 95.2 (84-100, P = .01): no revision was undertaken for aseptic loosening or fixation failure. Considering reoperation and bearing revision as end points, survival rates were 95.8% and 100%, respectively. The mean blood concentrations of chromium, cobalt, and titanium were 1.85 MUg/L, 1.24 MUg/L, and 9.62 MUg/L, respectively. A longer follow-up is mandatory, but it seems possible to use hard-on-hard bearings with metallic rings in young patients during hip revisions or in dysplastic cases with encouraging intermediate follow-up results. PMID- 20022454 TI - Cup anteversion in hip resurfacing: validation of EBRA and the presentation of a simple clinical grading system. AB - The use of large metal on metal bearings has led to a reduction in the risk of dislocation post hip arthroplasty. Because of this, and also because of the technical difficulties associated with resurfacing surgery in particular, it could be argued that a less meticulous approach to acetabular cup placement has developed in comparison with conventional metal on polyethylene arthroplasty. Resurfacing cups may produce significant clinical problems when placed at the extremes of version, including increased production of metal debris and psoas tendonitis. Presented in this article is evidence that EBRA software (Einzel-Bild Roentgen-Analysis, University of Innsbruck, Austria) can be used to reliably assess the version of resurfacing cups, when radiographs are of sufficient quality. The cups have characteristic appearances when placed at the extremes of version. These characteristics can allow the surgeon to identify poorly positioned cups without the use of software. PMID- 20022455 TI - The position of the tibial component affecting the postoperative mechanical axis in total knee arthroplasty. AB - The purpose is to identify whether the position of the tibial component in relation to the anatomical axis affects the postoperative mechanical axis in total knee arthroplasty for Korean patients. Preoperatively, 30 patients with varus deformity lesser than 10 degrees were classified as group A, and 30 patients greater than 10 degrees were classified as group B. Postoperatively, the distance between the midline of the tibial stem and anatomical axis (medial offset) was measured. The medial offsets were 2.5+/-1.9 mm in group A and 3.9+/ 2.7 mm in group B (P=.021). The postoperative mechanical axes were varus 1.3+/ 1.2 degrees in group A and varus 2.5+/-2.0 degrees in group B (P=.004). We think that the medial position of tibial component in relation to the anatomical axis affects the measurement of postoperative mechanical axis in total knee arthroplasty. PMID- 20022456 TI - Common errors in the execution of preoperative templating for primary total hip arthroplasty. AB - We reviewed 75 primary total hip arthroplasty preoperative and postoperative radiographs and recorded limb length discrepancy, change in femoral offset, acetabular position, neck cut, and femoral component positioning. Interobturator line, as a technique to measure preoperative limb length discrepancy, had the least amount of variance when compared with interteardrop and intertuberosity lines (Levene test, P = .0527). The most common error in execution of preoperative templating was excessive limb lengthening (mean, 3.52 mm), primarily due to inferior acetabular cup positioning (Pearson correlation coefficient, P = .036). Incomplete medialization of the acetabular component contributed the most to offset discrepancy. The most common errors in the execution of preoperative templating resulted in excessive limb lengthening and increased offset. Identifying these errors can lead to more accurate templating techniques and improved intraoperative execution. PMID- 20022457 TI - Efficacy of periarticular injection of bupivacaine, fentanyl, and methylprednisolone in total knee arthroplasty:a prospective, randomized trial. AB - We evaluated the efficacy of periarticular infiltration of corticosteroid, opioid, and a local anesthetic by comparing pain scores, knee flexion, and quadriceps function on the day of surgery, first postoperative day, day of discharge, and 2 and 4 weeks after surgery between the infiltrated and the noninfiltrated knee in 40 patients undergoing simultaneous bilateral computer assisted total knee arthroplasty who were randomized to receive the injection in the right or left knee. In comparison to the noninfiltrated side, the infiltrated knee showed significantly lower pain scores, significantly greater active flexion up to 4 weeks, and superior quadriceps recovery up to 2 weeks after surgery. This simple and inexpensive technique can significantly reduce pain and hasten functional recovery in the first month after total knee arthroplasty. PMID- 20022458 TI - Unexpected finding of a fractured metal prosthetic femoral head in a nonmodular implant during revision total hip arthroplasty. AB - Though there are many reports of fractured femoral components after total hip arthroplasty; there are no reports of a fractured metal femoral head. This is a report of a fractured metal femoral head in a nonmodular total hip replacement discovered unexpectedly during revision total hip arthroplasty for acetabular failure. This surprise finding, which was not appreciated on preoperative x-rays, required unplanned extraction of a well-ingrown, fully porous coated cylindrical femoral stem. Though rare, fracture of the metal femoral head in a DePuy (Warsaw, Ind.) monoblock Anatomic Medullary Locking (AML) component is possible, and one cannot expect the fracture to be apparent on preoperative radiographs as the 2 pieces may not necessarily dissociate. As usual, the surgeon performing revision arthroplasty should be prepared to revise all components. PMID- 20022459 TI - Using VistA electronic medical record data extracts to calculate the waiting time for total knee arthroplasty. AB - This is a retrospective database study of veterans who had total knee arthroplasty (TKA) at Veterans Affairs Connecticut Healthcare System. The objective of this study is to determine if VistA medical records data can be used to create a methodology for accurate assessment of waiting times for TKAs performed at Veterans Affairs facilities. The average waiting period from date of "initial consult" to date TKA was performed was greater than two years. The average waiting period from "most recent consult" to TKA was less than a year. This new approach and methodology has great impact as it provides an electronic method for calculating the TKA wait time which is broadly generalizable for similar analysis at the VISN (Veteran Integrated Services Network) or regional level. PMID- 20022460 TI - Randomized controlled trial of radiographic and patient-assessed outcomes following fixed versus rotating platform total knee arthroplasty. AB - This prospectively blinded randomized controlled study evaluated the difference in the functional and radiological outcomes in patients who received a press-fit condylar Sigma cemented cruciate-substituting total knee arthroplasty with either a rotating platform (RP) or a fixed bearing (FB). There were 51 joints in 49 patients: 24 joints in the RP group (mean follow-up, 43 months) and 27 joints in the FB group (mean follow-up, 40 months). At baseline, there was no significant difference in age, body mass index, preoperative diagnosis, Charnley class, range of motion, clinical and functional scores, between the RP and FB groups. At mid term follow up both the RP and FB give equivalent results, but patients with the RP tended to have a higher activity level score. PMID- 20022461 TI - High outpatient pain intensity scores predict impending hospital admissions in patients with cancer. AB - CONTEXT: Pain intensity scores (PIS) are frequently collected in the outpatient setting. The implications for patients with high PIS have not been well-studied. OBJECTIVES: This retrospective review was designed to determine whether high outpatient encounter PIS identify patients at risk of hospital admission. METHODS: Numerical PIS (0-10) were collected from all outpatient medical and radiation oncology encounters at the Johns Hopkins Comprehensive Cancer Center from 2004 to 2006. These were merged with an inpatient database to identify admissions occurring within 30 days of the outpatient encounter. PIS were categorized as 0-3 (mild), 4-6 (moderate), and 7-10 (severe). Odds ratios for hospital admission were calculated using generalized estimating equations. RESULTS: Of 119,069 encounters, 116,713 (98%) were evaluable, and 5,089 encounters (4.5%) had PIS of 7-10. Twenty-nine percent of these high PIS encounters had hospital admissions within 30 days. Encounters with PIS of 7-10 and 4-6 were 96% and 43%, respectively, more likely to result in hospital admission within 30 days compared with encounters with PIS<4 (P<0.001). Hospital admission rates after encounters with PIS of 7-10 were highest in patients with melanoma (58%), sarcoma (42%), female genital cancer (39%), and upper aerodigestive (36%) cancer. CONCLUSION: Outpatients with cancer and high PIS are at increased risk of hospital admission within 30 days. This high-risk group should be targeted for early supportive care interventions aimed at reducing hospitalizations and improving quality of life. PMID- 20022462 TI - An evaluation of Bayesian techniques for controlling model complexity and selecting inputs in a neural network for short-term load forecasting. AB - Artificial neural networks have frequently been proposed for electricity load forecasting because of their capabilities for the nonlinear modelling of large multivariate data sets. Modelling with neural networks is not an easy task though; two of the main challenges are defining the appropriate level of model complexity, and choosing the input variables. This paper evaluates techniques for automatic neural network modelling within a Bayesian framework, as applied to six samples containing daily load and weather data for four different countries. We analyse input selection as carried out by the Bayesian 'automatic relevance determination', and the usefulness of the Bayesian 'evidence' for the selection of the best structure (in terms of number of neurones), as compared to methods based on cross-validation. PMID- 20022463 TI - Delay-dependent stability analysis for continuous-time BAM neural networks with Markovian jumping parameters. AB - This paper investigates the problem of stability analysis for bidirectional associative memory (BAM) neural networks with Markovian jumping parameters. Some new delay-dependent stochastic stability criteria are derived based on a novel Lyapunov-Krasovskii functional (LKF) approach. These new criteria based on the delay partitioning idea prove to be less conservative, since the conservatism could be notably reduced by thinning the delay partitioning. It is shown that the addressed stochastic BAM neural networks with Markovian jumping parameters are stochastically stable if three linear matrix inequalities (LMIs) are feasible. The feasibility of the LMIs can be readily checked by the Matlab LMI toolbox. A numerical example is provided to show the effectiveness and advantage of the proposed technique. PMID- 20022464 TI - Effects of fruit consumption on body mass index and weight loss in a sample of overweight and obese dieters enrolled in a weight-loss intervention trial. AB - OBJECTIVE: Fruit and vegetable consumption is an important part of a healthy diet and assumed to aid in the reduction of energy intake and body weight. Fruits and vegetables may display differential effects on weight and weight loss; however, the effects of the two food groups have rarely been investigated separately. METHODS: The present study focused on the effects of fruit consumption on body weight and weight loss in a sample of 77 overweight and obese dieters enrolled in an intervention program. Food consumption was assessed at baseline by food diaries and after the introduction of nutrition software through electronic food records. Body weight and additional physiologic outcomes were assessed three times, once before the intervention and again at the 3- and 6-mo follow-ups. RESULTS: Vegetable and fruit consumption differed in their associations with body weight and weight loss. Although vegetable consumption increased as a result of the intervention (P<0.01), fruit consumption did not. However, only fruit consumption was associated with body mass index, showing an inverse relation with body weight in cross-sectional and longitudinal analyses (r=-0.27 to -0.44). The relation between fruit consumption and body weight remained significant after controlling for age, gender, physical activity level, and daily macronutrient consumption (DeltaR(2)=0.06-0.13). Further, increases in fruit consumption were associated with subsequent weight loss, controlling for the same covariates (DeltaR(2)=0.05-0.07). CONCLUSION: The results indicate unique contributions of fruit consumption to the management of body weight and indicate that a separation of effects for fruit and vegetable food groups may be warranted. PMID- 20022465 TI - Obesity: genes, brain, gut, and environment. AB - Obesity, which is assuming alarming proportions, has been attributed to genetic factors, hypothalamic dysfunction, and intestinal gut bacteria and an increase in the consumption of energy-dense food. Obesity predisposes to the development of type 2 diabetes mellitus, hypertension, coronary heart disease, and certain forms of cancer. Recent studies have shown that the intestinal bacteria in obese humans and mice differ from those in lean that could trigger a low-grade systemic inflammation. Consumption of a calorie-dense diet that initiates and perpetuates obesity could be due to failure of homeostatic mechanisms that regulate appetite, food consumption, and energy balance. Hypothalamic factors that regulate energy needs of the body, control appetite and satiety, and gut bacteria that participate in food digestion play a critical role in the onset of obesity. Incretins, cholecystokinin, brain-derived neurotrophic factor, leptin, long-chain fatty acid coenzyme A, endocannabinoids and vagal neurotransmitter acetylcholine play a role in the regulation of energy intake, glucose homeostasis, insulin secretion, and pathobiology of obesity and type 2 diabetes mellitus. Thus, there is a cross-talk among the gut, liver, pancreas, adipose tissue, and hypothalamus. Based on these evidences, it is clear that management of obesity needs a multifactorial approach. PMID- 20022466 TI - Effect of 6-month nutritional intervention on non-alcoholic fatty liver disease. AB - OBJECTIVE: We evaluated the effect of diet therapy as exclusive treatment on insulin resistance, biochemical parameters of metabolic syndrome, and degree of hepatic steatosis in patients with non-alcoholic fatty liver disease. METHODS: Thirty-one patients with non-alcoholic fatty liver disease received a diet with a reduction of 500 to 1000 cal/d, containing 15% protein, 55% carbohydrates, and 30% fat, for 6 mo. At entry and 6 mo after dietary instructions, degrees of hepatic steatosis and visceral obesity were assessed by computed tomography; serum aspartate aminotransferase, alanine aminotransferase, gamma-glutamyl transferase, glucose, triacylglycerols, and high-density lipoprotein cholesterol were measured by automated methods. Body mass index, waist circumference, waist to-hip ratio, and food intake (7-d diary) were also evaluated. At the end of follow-up, the patients were classified as adherent or non-adherent to treatment according to a weight loss of more or less than 5% of initial body weight, respectively. RESULTS: Seventeen patients were classified as adherent (group 1) and 14 as non-adherent (group 2). Group 2 only presented a significant reduction in body mass index and waist circumference. In contrast, in group 1, in addition to significant improvement of all anthropometric parameters, a significant reduction was observed in alanine aminotransferase and gamma-glutamyl transferase levels, homeostasis model assessment for insulin resistance, visceral fat and tomographic liver density, together with an increase in serum high-density lipoprotein cholesterol levels. These patients presented a significant decrease in total energy intake and in total and saturated fats. CONCLUSION: Nutritional intervention as exclusive treatment, with a loss of at least 5% of initial weight, is effective in the treatment of non-alcoholic fatty liver disease. PMID- 20022467 TI - Estimation of protein loss from wound fluid in older patients with severe pressure ulcers. AB - OBJECTIVE: Protein loss from wound fluid is usually recognized as one of the factors contributing to the deterioration of the nutritional status in older patients with severe pressure ulcers. We quantified the protein loss owing to pressure ulcers and investigated associations with wound-related factors and nutritional status. METHODS: This cross-sectional study included 25 patients (>or=60 y) from 10 institutions, with full-thickness pressure ulcers. Wound fluid was collected once after accumulating beneath a film dressing. The amount of protein loss per day was estimated by the volume of wound fluid per hour and the total protein concentration in the wound fluid. Wound evaluations and nutritional assessments were performed. Correlations between variables were obtained using Spearman's rank correlation. RESULTS: The median age of the patients was 79 y (range 61-100), and median body mass index was 19.6 kg/m(2) (12.2-24.9). The median amount of protein loss was 0.2g/d (0.04-2.1), which corresponded to 0.01 g x kg(-1) x d(-1) (<0.01-0.04) and 0.6% (0.1-13.8) of protein intake. Four wounds characterized as infected or surgically debrided lost 1.5-2.1g of protein per day, which was substantially higher than other wounds lost. Protein loss was correlated with wound severity including area, depth, the wound severity score, and infectious markers (all Ps<0.05), but not with body mass index or arm muscle circumference (P>0.05). CONCLUSION: The amount of protein loss could be small and thus may not be related directly to nutritional status, although it increased as the wound became more severe. PMID- 20022468 TI - Diet supplementation with acai (Euterpe oleracea Mart.) pulp improves biomarkers of oxidative stress and the serum lipid profile in rats. AB - OBJECTIVE: We investigated the antioxidant potential and hypocholesterolemic effects of acai (Euterpe oleracea Mart.) pulp ingestion in rats fed a standard or hypercholesterolemic diet. METHODS: Female Fischer rats were fed a standard AIN 93 M diet (control) or a hypercholesterolemic diet that contained 25% soy oil and 1% cholesterol. The test diet was supplemented with 2% acai pulp (dry wt/wt) for control (group CA) and hypercholesterolemic rats (group HA) for 6 wk. At the end of the experimental period, rats were sacrificed and the blood and livers were collected. To evaluate the effect of acai consumption, levels of protein carbonyl and sulfhydryl groups, superoxide dismutase and paraoxonase activities, and lipid profiles of the sera were measured. RESULTS: Animals that were fed the hypercholesterolemic diet presented increased levels of total and non-high density lipoprotein cholesterol and decreased levels of high-density lipoprotein cholesterol. Supplementing the diet of this group with acai caused a hypocholesterolemic effect by reducing total and non-high-density lipoprotein cholesterol. Serum levels of carbonyl proteins and total, free, and protein sulfhydryl groups were reduced by acai ingestion in animals receiving the standard or hypercholesterolemic diet. Acai supplementation induced a significant reduction in superoxide dismutase activity only in the hypercholesterolemic rats, indicating an association between diet and acai treatment. Also, acai supplementation increased paraoxonase activity in the CA and HA groups. CONCLUSION: These results suggest that the consumption of acai improves antioxidant status and has a hypocholesterolemic effect in an animal model of dietary-induced hypercholesterolemia. PMID- 20022469 TI - Effects of different ratios of monounsaturated and polyunsaturated fatty acids to saturated fatty acids on regulating body fat deposition in hamsters. AB - OBJECTIVE: Effects of monounsaturated fatty acid (MUFA) and polyunsaturated fatty acid consumption on regulating body fat accumulation and body weight gain are controversial between animal and human studies. METHODS: We designed a 2 x 2 factorial study, with two levels of MUFAs (60% and 30%) and two levels of polyunsaturated-to-saturated fatty acid (P/S) ratio (5 and 3) to prepare four kinds of experimental oils consisting of 60% MUFAs with a high or low P/S ratio (HMHR or HMLR, respectively) or 30% MUFAs with a high or low P/S ratio (LMHR or LMLR, respectively). Thirty-two male golden Syrian hamsters were randomly divided into four groups and fed the experimental diets containing 15% (w/w) fat for 12 wk. RESULTS: No difference was observed in the mean daily food intake. Hamsters fed the LMLR diet had increased weight gain, epididymal and retroperitoneal white adipose tissues, plasma non-esterified fatty acids, insulin, hepatic acetyl coenzyme A carboxylase and malic enzyme activities, and mRNA expressions of peroxisome proliferator-activated receptor-alpha and sterol regulatory element binding protein-1c among all groups (P < 0.05). Hamsters fed the HMHR diet had lower plasma insulin levels and hepatic acetyl coenzyme A carboxylase activities among groups (P < 0.05) and elevated hepatic acyl coenzyme A oxidase and carnitine palmitoyltransferase-I activities compared with those fed the LMLR diet (P < 0.05). CONCLUSION: Hamsters fed the LMLR diet had increased weight gain and body fat accumulation, whereas the HMHR diet appeared to be beneficial in preventing white adipose tissue accumulation by decreasing plasma insulin levels and increasing hepatic lipolytic enzyme activities involved in beta-oxidation. PMID- 20022470 TI - Intranasal insulin: from nose to brain. AB - BACKGROUND: Intranasal insulin has proven useful to control hyperglycemia in diabetics but its mechanism of action has not been well defined. We attempted to understand several aspects of human insulin metabolism by measurement of and interaction of insulin and its associated moieties in nasal mucus, saliva and blood plasma under various physiological and pathological conditions. METHODS: Insulin, insulin receptors, insulin-like growth factor 1 (IGF1) and insulin-like growth receptor 3 (IGFR3) were measured in nasal mucus, saliva and blood plasma in normal subjects, in thin and obese subjects and in diabetics under fasting and fed conditions. RESULTS: There are complex relationships among each of these moieties in each biological fluid. Insulin and its associated moieties are present in both nasal mucus and saliva. These moieties in nasal mucus and saliva report on physiological and pathological changes in glucose metabolism as do these moieties in plasma. Indeed, insulin and its associated moieties in nasal mucus may offer specific data on how insulin enters the brain and thereby play essential roles in control of insulin metabolism. INTERPRETATION: These data support the concept that insulin is synthesized not only in parotid glands but also in nasal serous glands. They also support the concept that insulin enters the brain following intranasal administration either 1) by direct entry through the cribriform plate, along the olfactory nerves and into brain parenchyma, 2) by entry through specific receptors in blood-brain barrier and thereby into the brain or 3) some combination of 1) and 2). Conversely, data also show that insulin introduced directly into the brain is secreted out of brain into the peripheral circulation. Data in this study demonstrate for the first time that insulin and its associated moieties are present not only in saliva but also in nasal mucus. How these complex relationships among nasal mucus, saliva and plasma occur are unclear but results demonstrate these relationships play separate yet interrelated roles in physiology and pathology of human insulin metabolism. PMID- 20022471 TI - MRS-lateralisation index in patients with epilepsy and focal cortical dysplasia or a MEG-focus using bilateral single voxels. AB - PURPOSE: To evaluate if single voxel proton magnetic resonance spectroscopy (SV MRS) can help in lateralising and sometimes in localizing an epileptogenic focus. The assumption is that in MRI negative patients the underlying pathology most often is focal cortical dysplasia (FCD). Several studies have shown that in the presence of FCD there are also (1)H-MRS abnormalities on the contralateral side. However, in most cases the studied group was not homogeneous and included different forms of dysplasias, including band heterotopias and polymicrogyria, and the studies used different spectroscopy protocols. In the present study, using bilateral SV-MRS we investigated the presence of a lateralisation index in two groups of patients with localisation related epilepsy: patients with focal cortical dysplasia on MRI and patients without MRI abnormalities with a focus identified by MEG. Aim of the study was to show that in both groups the expected epileptogenic side shows more pronounced metabolic alterations, making MRS a possible screening tool for clarifying lateralisation questions in patients with cryptogenic localisation related epilepsy. METHODS: In ten patients a single voxel was placed over the FCD and in nine patients over the region of interest (ROI) as indicated by MEG. In all patients a voxel was also placed in the contralateral homologus location. We used metabolite concentrations as peak ratios relative to the creatine (Cr) peak to calculate a lateralisation index. RESULTS: In both groups NAA/Cr was significantly lower on the affected side whereas the results for Cho/Cr were more diverse. There were no significant differences between the two groups. The limitations of the used methods and the implications of the findings are discussed. PMID- 20022472 TI - Epidermal triglyceride levels are correlated with severity of ichthyosis in Dorfman-Chanarin syndrome. AB - BACKGROUND: Dorfman-Chanarin syndrome (DCS), also referred to as neutral lipid storage disease with ichthyosis, is a rare autosomal recessive form of nonbullous congenital ichthyosiform erythroderma, characterized by the presence of intracellular lipid droplets in multiorgans. DCS patients often have mutations in CGI-58, which is an activator of adipose triglyceride lipase (ATGL), leading to accumulation of triglycerides (TG). OBJECTIVE: To study whether a patient with DCS demonstrates TG accumulation in the epidermis and to analyze whether TG levels are correlated with skin disease activity. METHODS: Skin specimen from a 62-year-old man with DCS was stained with oil red O, and analyzed on electromicrographs. Sequencing analysis of CGI-58 was performed using the patient's blood cells. The scales from the lesion were subject to lipid analysis by high-performance thin-layer chromatography (HPTLC). RESULTS: The patient demonstrated ichthyoform erythroderma with a distinct seasonal fluctuation: his skin lesions were aggravated in summer but resolved during winter. Epidermis of the lesion showed intracellular lipid droplets. Sequencing analysis revealed a novel missense mutation in the exon 3 of CGI-58 gene. Lipid analysis of the scales from his lesions, compared with those from normal human control, revealed increased levels of triglycerides (TG) but, in turn, decreased levels of free fatty acids, suggesting dysfunction of adipose TG lipase. Notably, the TG levels in the scales from the patient were positively correlated with the severity of ichthyosis. CONCLUSION: These results suggest that TG accumulation by epidermal keratinocytes directly contributes to ichthyosiform phenotype of DCS. PMID- 20022474 TI - Evaluation of azithromycin, trovafloxacin and grepafloxacin as prophylaxis against experimental murine Brucella melitensis infection. AB - The prophylactic potential of the azalide azithromycin as well as the fluoroquinolones trovafloxacin and grepafloxacin was assessed for the control of infection with Brucella melitensis in an experimental mouse model, determined by reduction in splenic bacterial burden. Trovafloxacin showed limited protective efficacy when administered 2h following a low-dose B. melitensis challenge, whereas grepafloxacin was ineffective. In comparison, azithromycin provided significant control of infection both following low- and high-dose challenges. Overall, the data confirm the potential utility of azithromycin in the prophylaxis of brucellosis and suggest that neither trovafloxacin nor grepafloxacin would likely be valuable for post-exposure prophylaxis of Brucella infection. PMID- 20022475 TI - NMDA NR2B subtype-selective receptor antagonists fail to antagonize electrically precipitated seizures and elicit popping in mice. AB - NR2B-subtype-selective antagonists differ from MK-801, a nonselective NMDA receptor antagonist. MK-801 antagonizes electrical seizures at doses as low as 0.1 to 0.18mg/kg and elicits popping at doses as low as 0.5mg/kg, whereas ifenprodil and Ro 8-4304 were unable to do so at the doses tested. Ro 25-6981, however, was able to antagonize electrically-precipitated tonic hindlimb extension at 100mg/kg, but was not able to elicit popping behavior at this dose. PMID- 20022473 TI - Review of hair follicle dermal cells. AB - Hair follicle stem cells in the epithelial bulge are responsible for the continual regeneration of the hair follicle during cycling. The bulge cells reside in a niche composed of dermal cells. The dermal compartment of the hair follicle consists of the dermal papilla and dermal sheath. Interactions between hair follicle epithelial and dermal cells are necessary for hair follicle morphogenesis during development and in hair reconstitution assays. Dermal papilla and dermal sheath cells express specific markers and possess distinctive morphology and behavior in culture. These cells can induce hair follicle differentiation in epithelial cells and are required in hair reconstitution assays either in the form of intact tissue, dissociated freshly prepared cells or cultured cells. This review will focus on hair follicle dermal cells since most therapeutic efforts to date have concentrated on this aspect of the hair follicle, with the idea that enriching hair-inductive dermal cell populations and expanding their number by culture while maintaining their properties, will establish an efficient hair reconstitution assay that could eventually have therapeutic implications. PMID- 20022476 TI - [Reflections on academic appointments in pediatrics]. PMID- 20022477 TI - Association of the CCR3 gene polymorphism with aspirin exacerbated respiratory disease. AB - INTRODUCTION: Aspirin hypersensitivity represents two distinct clinical syndromes, such as aspirin exacerbated respiratory disease (AERD) and aspirin intolerant chronic urticaria/angioedema (AICU) which have different clinical phenotypes resulting from different genetic backgrounds in a Korean population. Persistent eosinophilic inflammation in airway is a characteristic feature of AERD and chemokine CC motif receptor 3 (CCR3) plays an important role in eosinophilic infiltration into the asthmatic airway. OBJECTIVES: The main objective of this study is to investigate the association between CCR3 gene polymorphisms and aspirin hypersensitivity, including AERD and AICU. METHODS: CCR3 mRNA expression was measured after an aspirin provocation test by real-time PCR. In total, 330 patients with aspirin hypersensitivity (191 AERD and 139 AICU) and 217 normal healthy controls (NC) were genotyped for two CCR3 promoter polymorphisms (-520T/G and -174C/T), and the functional effects of the polymorphisms were analyzed applying a luciferase reporter assay and an electrophoretic mobility shift assay. RESULTS: CCR3 mRNA expression was significantly increased after aspirin provocation in AERD patients (P=0.002) but not in AICU patients. An in vitro functional study showed that the reporter construct having a -520G allele exhibited significantly higher promoter activity compared with the construct having a -520T allele in human myeloid (U937), lymphoid (Jurkat), and mast (HMC-1) cell lines (P<0.001). We found -520G and 174T specific bands on EMSA. CONCLUSION: This result suggests that the CCR3 genetic polymorphisms may contribute to the development of the AERD phenotype and may be used as a genetic marker for differentiating between the two major aspirin hypersensitivity phenotypes. PMID- 20022478 TI - A comparison of municipal solid waste management in Berlin and Singapore. AB - A comparative analysis of municipal solid waste management (MSWM) in Singapore and Berlin was carried out in order to identify its current status, and highlight the prevailing conditions of MSWM. An overview of the various aspects of MSWM in these two cities is provided, with emphasis on comparing the legal, technical, and managerial aspects of MSW. Collection systems and recycling practiced with respect to the involvement of the government and the private sector, are also presented. Over last two decades, the city of Berlin has made impressive progress with respect to its waste management. The amounts of waste have declined significantly, and at the same time the proportion that could be recovered and recycled has increased. In contrast, although Singapore's recycling rate has been increasing over the past few years, rapid economic and population growth as well as change in consumption patterns in this city-state has caused waste generation to continue to increase. Landfilling of MSW plays minor role in both cities, one due to geography (Singapore) and the other due to legislative prohibition (Berlin). Consequently, both in Singapore and Berlin, waste is increasingly being used as a valuable resource and great efforts have been made for the development of incineration technology and energy recovery, as well as climate protection. PMID- 20022480 TI - Improved performance of single-chamber microbial fuel cells through control of membrane deformation. AB - Cation (CEMs) and anion exchange membrane (AEMs) are commonly used in microbial fuel cells (MFCs) to enhance Coulombic efficiencies (CEs) by reducing the flux of oxygen through the cathode to bacteria on the anode. AEMs typically work better than CEMs, but in initial experiments we observed the opposite using a membrane electrode assembly MFC. The reason was identified to be membrane deformation, which resulted in water and gas trapped between the membrane and cathode. To correct this, stainless steel mesh was used to press the membrane flat against the cathode. With the steel mesh, AEM performance increased to 46+/-4 W/m(3) in a single cathode MFC, and 98+/-14 W/m(3) in a double-cathode MFC. These power densities were higher than those using a CEM of 32+/-2 W/m(3) (single cathode) and 63+/-6 W/m(3) (double cathode). Higher pH gradients across the membrane and salt precipitation on the cathode were responsible for the reduced performance of the CEM compared to the AEM. CEs reached over 90% for both membranes at >2A/m(2). These results demonstrate the importance of avoiding water accumulation in thin films between membranes and electrodes, and explain additional reasons for poorer performance of CEMs compared to AEMs. PMID- 20022479 TI - Flow tracing microparticle sensors designed for enhanced X-ray contrast. AB - In applying the X-ray particle image velocimetry (PIV) technique to biofluid flows, the most pivotal prerequisite is suitable flow tracing sensors which should be detected effectively by the X-ray imaging system. In this study, to design those flow tracing sensors, X-ray contrast agent Iopamidol was encapsulated into the poly(vinyl alcohol) (PVA) microparticles crosslinked by glutaraldehyde (GA). The characteristics of the fabricated particle sensors were determined by optical microscopy, scanning electron microscopy, dynamic light scattering, laser Doppler electrophoresis and nuclear magnetic resonance spectroscopy ((1)H NMR). The amount of Iopamidol in the microparticles was measured using the energy dispersive X-ray spectroscopy (EDS) and (1)H NMR. The physical properties of the PVA microparticles are effectively controlled in terms of the average particle size, degree of crosslinking, degree of swelling and encapsulation efficiency of Iopamidol. By changing the amount of crosslinker, the degree of crosslinking and the efficiency of the Iopamidol encapsulation reached to the optimal. To some extent, the zeta-potential of the PVA microparticles is increased in less ionic media where the particles can effectively repel each other prohibiting aggregation. The X-ray absorption ability of the designed tracing sensors was examined by a synchrotron X-ray imaging technique. The X-ray absorption coefficients of the particle sensors were expressed by an exponential law assuming the spherical shape of the microparticles. The X-ray contrast agent, Iopamidol, was successfully encapsulated into the bio-compatible and bio degradable PVA. With the controlled physical properties of the flow tracing sensors designed in this study, the particle sensors exhibit excellent X-ray absorption contrast fairly applicable in biological systems. PMID- 20022481 TI - Rapid and direct magnetization of GFP-reporter yeast for micro-screening systems. AB - Saccharomyces cerevisiae containing fluorescent markers are ideal candidates for applications in microfluidic screening systems as fluorescence signal is emitted without the need of additional reagents. Here we develop a method for magnetic functionalization of such cells which allows their handling and immobilization in micro-screening devices. After exposure of the magnetized GFP-reporter yeast (GreenScreen) to a genotoxic compound, the fluorescence emission was detected using fluorescent spectrometer and an Epi-fluorescent microscope. Results demonstrate that GFP production and fluorescence emission is not altered by their magnetic functionalization, indicating its potential employment on biosensors, bioreactors and micro-screening studies. PMID- 20022482 TI - Carbon nanotubes with platinum nano-islands as glucose biofuel cell electrodes. AB - A novel method using intense pulsed light (IPL) for the metal nano-island formation on carbon nanotube (CNT) was introduced. The IPL-induced photothermal dewetting process improved platinum (Pt) catalyst utilization by transforming nano-islands from Pt film on CNT and increasing the surface area for the subsequent sputtering. The irradiation of high intensity of light on the Pt film causes surface-energy-driven diffusion of Pt atoms and forms the array of nano islands on CNT. The thickness of Pt film can change the size of nano-islands. Cyclic voltammetry showed a dramatically improved glucose oxidation at the IPL morphology modified Pt-CNT electrode compared to the Pt sputtered CNT electrode without IPL irradiation. The power densities of glucose/air biofuel cell based on the morphology modified Pt-CNT electrode and the as-sputtered Pt-CNT electrode were 0.768 microW/cm(2) and 0.178 microW/cm(2), respectively. The biofuel cell based on morphology modified Pt-CNT electrode showed highly stable output in long term performance. The power density dropped 14.1% in 30 days. Efforts are underway to improve the interface transfer to achieve higher potential and current output. PMID- 20022483 TI - Anodized aluminum oxide-based capacitance sensors for the direct detection of DNA hybridization. AB - We fabricated a capacitance sensor based on an anodized aluminum oxide (AAO) nanoporous structure to detect DNA hybridization. We utilized Au film deposited on the surface of the AAO membrane and Au nanowires infiltrating the nanopores as the top and bottom electrodes, respectively. When completely complementary target DNA molecules were added to the sensor-immobilized DNA molecule probes, the capacitance was reduced; with a concentration of 1pM, the capacitance decreased by approximately 10%. We measured the capacitance change for different concentrations of the target DNA solution. A linear relationship was found between the capacitance change and DNA concentration on a semi-logarithmic scale. We also investigated the possibility of detecting DNA molecules with a single base mismatch to the probe DNA molecule. In contrast to complementary target DNA molecules, the addition of one-base mismatch DNA molecules caused no significant change in capacitance, demonstrating that DNA hybridization was detected with single nucleotide polymorphism sensitivity. PMID- 20022484 TI - A sensitive fluorescence anisotropy method for the direct detection of cancer cells in whole blood based on aptamer-conjugated near-infrared fluorescent nanoparticles. AB - Based on the aptamer-conjugated core-shell near-infrared fluorescent nanoparticles (NIR-Nps) and fluorescence anisotropy measurement, the present study reported proof-of-principle for a rapid homogeneous assay approach that can detect target cancer cells without the need of the complicated separation steps in whole blood samples. Experimental investigation showed that the novel NIR-Nps have negligible background fluorescence and low inner filtration interference in complex biologic systems such as whole blood. The specific recognition characteristic of aptamer in whole blood samples was investigated by using the proposed fluorescence anisotropy method. The results showed that the fluorescent nanoparticle-tagged aptamer probes sequence could achieve specific recognition of the target cancer cells from complex mixtures including whole blood samples. And the reaction conditions for the binding between fluorescent nanoparticle conjugated aptamer probes and target cancer cells were optimized. The present approach can exhibit sensitive and reproducible fluorescence anisotropy responses to the target cells concentration and the calibration curve showed good linearity when the target cells concentration is in the range from 4.0 x 10(3) to 7.0 x 10(5)cells/mL. Moreover, the present fluorescence anisotropy assay technique could be practically utilized for the detection of acute leukemia samples with improved capabilities and be comparable to the immunophenotyping methods clinically used. PMID- 20022485 TI - The evolution of real-time PCR machines to real-time PCR chips. AB - Development of Micro-Electro-Mechanical Systems (MEMS) technology has recently allowed the migration of real-time polymerase chain reaction (PCR) machines to lab-on-a-chip systems. The miniaturization of biological instruments has been studied extensively, with several prototypes constructed and tested. In this study, the lab-on-a-chip system is evaluated; its DNA quantification is estimated by theorems, and the specifications of proposed chip prototypes are compared with the original machines. The analysis results suggest five hypotheses. Using experiments and the data collected from published papers, these hypotheses were either verified or rejected, and the advantages and shortcomings of real-time PCR chips were identified. The proven advantages of the lab-on-a-chip system are its compact size, low sample volume to nano-liter, and short analysis time-less than 10s to complete one PCR cycle and 370 s for completing the whole quantification process. However, the detection limits, quantification uncertainties, and melting analysis ability of the chip prototypes are at best comparable to, and perhaps worse than, those of commercial instruments. Real-time PCR chips are not perfectly accurate diagnostic tools for a laboratory but they have advantages over traditional techniques for point-of-care testing. PMID- 20022486 TI - Prognostic impact of thymidine phosphorylase expression in breast cancer- comparison of microarray and immunohistochemical data. AB - Contrary findings exist according to the prognostic and predictive impact of thymidine phosphorylase (TP) expression in breast cancer. Goal of our study was to investigate TP expression on the mRNA level by microarray analysis in a large cohort of 1781 breast cancers and to analyse its prognostic impact. Furthermore we compared mRNA expression and immunohistochemical data to explain discrepancies between different studies. The prognostic value of TP mRNA expression was analysed among n=622 untreated patients. Strong expression in the subgroup of n=213 ER-negative cancer correlates with improved survival (P=0.012). In contrast, no difference in survival was detected in the ER-positive group. We also failed to observe a prognostic value of TP mRNA among n=435 endocrine treated patients as well as n=111 CMF-treated patients. In an unsupervised analysis, TP clustered together with genes expressed in immune cells. Moreover, among normal tissues the highest TP mRNA expression was found in tissues of the immune system. The profile of TP expression in breast cancers correlates to a metagene of interferon induction whereas the expression of TP among normal tissues correlates to a metagene for macrophages. When comparing microarray data with immunohistochemistry from the same n=51 samples, there was no correlation with stained carcinoma cells. In contrast, the correlation with stromal staining was highly significant (P<0.001). Thus TP mRNA from microarray mainly reflects expression in stromal and immune cells. This could account for discrepant results from mRNA and IHC studies. In conclusion, the tumour infiltrating immune cells seem to be a major source of TP expression and predict a favourable prognosis in ER-negative breast cancer. Our data point to a role of TP in host immune response. PMID- 20022488 TI - Red algae and their use in papermaking. AB - Gelidialian red algae, that contain rhizoidal filaments, except the family Gelidiellaceae were processed to make bleached pulps, which can be used as raw materials for papermaking. Red algae consist of rhizoidal filaments, cortical cells usually reddish in color, and medullary cells filled with mucilaginous carbohydrates. Red algae pulp consists of mostly rhizoidal filaments. Red algae pulp of high brightness can be produced by extracting mucilaginous carbohydrates after heating the algae in an aqueous medium and subsequently treating the extracted with bleaching chemicals. In this study, we prepared paper samples from bleached pulps obtained from two red algae species (Gelidium amansii and Gelidium corneum) and compared their properties to those of bleached wood chemical pulps. PMID- 20022487 TI - Biological treatment of a dye solution by Macroalgae Chara sp.: effect of operational parameters, intermediates identification and artificial neural network modeling. AB - The potential of a macroalgae Chara sp. was investigated as a viable biomaterial for biological treatment of Malachite Green (MG) solution. The effects of operational parameters such as temperature, pH, initial dye concentration, reaction time and amount of algae on biological decolorization efficiency were studied. Biological treatment of MG solution by live and dead algae was compared. The reusability and efficiency of the live algae in long-term repetitive operations were also examined. The batch experiments results revealed the ability of algal species in biological degradation of the dye. The biological degradation compounds formed in this process were analyzed by UV-Vis, FT-IR and GC-Mass techniques. The degradation pathway of MG was proposed based on the identified compounds. In addition, an artificial neural network model was developed to predict the biological degradation efficiency. The findings indicated that ANN provides reasonable predictive performance (R(2)=0.970). The influence of each parameter on the variable studied was assessed, reaction time being the most significant factor, followed by temperature of the solution. PMID- 20022489 TI - Base-induced delignification of Miscanthus x giganteus studied by three dimensional confocal Raman imaging. AB - Confocal raman microscopy has been used to monitor the structural and chemical changes upon NaOH treatment of Miscanthus x giganteus, a potential energy crop and a model lignocellulosic material. Longitudinal and transversal-section images of the parenchyma cells in raw miscanthus samples reveal that lignin and cellulose are collocated in the cell wall and that a globular structure, composed predominantly of hemicellulose and lignin is associated with the interior cell wall. NaOH treatment results in the complete removal of lignin at long processing time but leaves the cellulose largely undisturbed as evidenced by the lack of conversion from type I to type II cellulose. Depth profiling images of partially processed (short exposure time) parenchyma cells reveal that lignin is removed preferentially from the interior surface of the cell wall as indicated by the anisotropic distribution of lignin and cellulose across the cell wall in partially processed samples. These spatially resolved chemical changes are important, because they illustrate how even simple pre-processing protocols can develop complex molecular profiles by differential rates of attack on the major components of the cell wall. PMID- 20022490 TI - Tolerance and removal of arsenic by a facultative marine fungus Aspergillus candidus. AB - The objective of the work was to investigate tolerance to and removal of arsenic by a facultative marine fungus Aspergillus candidus. The fungus showed luxuriant growth in different concentrations (25 and 50mg/L) of trivalent and pentavalent forms of arsenic. Biomass accumulation data substantiate tolerance of A. candidus towards the test concentrations of trivalent and pentavalent forms of arsenic. Highest arsenic removal (mg/g) was recorded on day 3. As removal increased with an increase in concentration. Hence, the test fungus A. candidus is a promising candidate for arsenic remediation. PMID- 20022491 TI - Nano-zerovalent iron contained porous carbons developed from waste biomass for the adsorption and dechlorination of PCBs. AB - The low-cost composite, nano-zerovalent iron (NZVI) contained in porous carbon (PC), was prepared using pinewood sawdust and ferric chloride as starting materials. The key point of this strategy was that the production of PC and the formation of NZVI were accomplished simultaneously through a simple process. The composite PC/NZVI was characterized by XRD, BET and the adsorption and simultaneous dechlorination of PCBs were efficiently demonstrated. The results showed the pinewood sawdust was activated by ferric chloride and the surface area and the pore volume of obtained composite were 423 m(2)/g and 0.23 cm(3)/g, respectively. The produced NZVI, around 27 nm in diameter, catalyzed the formation of substantial mesopores in the composite. PC/NZVI exhibited an efficient dechlorination of PCBs at room temperature, and the dechlorinated products could be completely adsorbed onto the composite. Accordingly, it is believed that PC/NZVI developed in the present study is practically applicable for PCBs-contaminated water purification. PMID- 20022492 TI - Design and synthesis of novel bis-oximinoalkanoic acids as potent PPAR alpha agonists. AB - Bis-oximinoalkanoic acid derivatives were designed and synthesized to aid in the characterization of selective PPARalpha agonists by replacing the oxazole ring with flexible oximino group in the lipophilic tail part of a previously reported compound 3. Selected compounds 9d and 9m showed excellent potency and high selectivity towards PPARalpha in vitro. These compounds found effective in reducing serum triglycerides (TG) in vivo. PMID- 20022493 TI - Heterocycle-substituted proline dipeptides as potent VLA-4 antagonists. AB - A variety of N-linked tertiary amines and heteroarylamines were examined at the 4 position of sulfonylated proline dipeptides in order to improve VLA-4 receptor off-rates and overcome the issue of CYP3A4 time-dependent inhibition of ester prodrugs. A tight-binding inhibitor 5j with a long off-rate provided sustained receptor occupancy despite poor oral pharmacokinetics. PMID- 20022494 TI - 2,3-Diaminopyrazines as Rho kinase inhibitors. AB - Inhibition of rho kinase (ROCK) has been recognized as an important target for a number of diseases, including glaucoma. Herein we report SAR development around two hits from a kinase library that led to the discovery of the ROCK inhibitor compound 38. In vitro and in vivo analysis of this compound, including its effects in a monkey model of glaucoma will be discussed. PMID- 20022495 TI - Natural ortho-dihydroxyisoflavone derivatives from aged Korean fermented soybean paste as potent tyrosinase and melanin formation inhibitors. AB - Natural o-dihydroxyisoflavone (ODI) derivatives with variable hydroxyl substituent at the aromatic ring of isoflavone and three known isoflavones were isolated from five-year-old Korean fermented soybean paste (Doenjang) and evaluated as potent inhibitors on tyrosinase activity and melanin formation in melan-a cells comparing with other known isoflavones, 7,8,4'-trihydroxyisoflavone (1) and 7,3',4'-trihydroxyisoflavone (2) inhibited tyrosinase by 50% at a concentration of 11.21+/-0.8 microM and 5.23+/-0.6 microM (IC(50)), respectively, whereas, 6,7,4'-trihydroxyisoflavone (3), daidzein (4), glycitein (5) and genistein (6) showed very low inhibition activity. Furthermore, those compounds significantly suppressed the cellular melanin formation by 50% at a concentration of 12.23+/-0.7 microM (1), 7.83+/-0.7 microM (2), and 57.83+/-0.5(6) and show more activity than arbutin. But, compounds 3, 4, and 5 showed lower inhibition activity. This study shows that the position of hydroxyl substituent at the aromatic ring of isoflavone plays an important role in the intracellular regulation of melanin formation in cell-based assay system. PMID- 20022496 TI - Simocyclinone D8 turns on against Gram-negative bacteria in a clinical setting. AB - Simocyclinone D8 (SD8) is known to affect Gram-positive bacteria only. By testing SD8 against several clinical isolates, we showed that SD8 resulted very active against Gram-negative bacteria from clinical specimens, while it was shown inactive against laboratory strains. The activity against the former was in part due to enhanced drug entry. In addition, SD8 appears to share chromosome- and plasmid-mediated resistance mechanisms with fluoroquinolones. PMID- 20022497 TI - 5'-O-dephosphorylated 2',5'-oligoadenylate (2-5A) with 8-methyladenosine at the 2'-terminus activates human RNase L. AB - Human ribonuclease L (RNase L), an interferon-induced endoribonuclease, becomes enzymatically active after binding to 2-5A. The 5'-phosphoryl group of 2-5A is reportedly necessary for the conformational change leading to RNase L activation. However, we found that 5'-O-dephosphorylated 2-5A tetramer analogs with 8 methyladenosine at the 2'-terminus were more effective as an activator of RNase L than the parent 2-5A tetramer. Introduction of 8-methyladenosine is thought to induce a dramatic shift of 2-5A in the binding site of RNase L. PMID- 20022498 TI - The discovery of novel tartrate-based TNF-alpha converting enzyme (TACE) inhibitors. AB - A novel series of TNF-alpha convertase (TACE) inhibitors which are non hydroxamate have been discovered. These compounds are bis-amides of L-tartaric acid (tartrate) and coordinate to the active site zinc in a tridentate manner. They are selective for TACE over other MMP's. We report the first X-ray crystal structure for a tartrate-based TACE inhibitor. PMID- 20022499 TI - Solution structural investigation and conformation-activity relationship of BAM8 22 by NMR and molecular dynamics simulations. AB - NMR spectroscopy and molecular dynamics simulations show that BAM8-22 (Val(8) Gly(9)-Arg(10)-Pro(11)-Glu(12)-Trp(13)-Trp(14)-Met(15)-Asp(16)-Tyr(17)-Gln(18) Lys(19)-Arg(20)-Tyr(21)-Gly(22)) possesses a relatively well-defined alpha-helix extending from Glu(12) to Arg(20), whereas both termini remain highly flexible in aqueous solution. The conformation-activity relationship of BAM8-22 indicates that the integrity of the well-defined alpha-helical structure is essential but not the sole determining factor for its bioactivity. PMID- 20022500 TI - Synthesis and antituberculosis activity of novel mefloquine-isoxazole carboxylic esters as prodrugs. AB - 5-(2,8-Bis(trifluoromethyl)quinolin-4-yloxymethyl)isoxazole-3-carboxylic acid ethyl ester (compound 3) was reported to have excellent antituberculosis activity against both replicating and non-replicating Mycobacterium tuberculosis, with a minimum inhibitory concentration (MIC) of 0.9 microM and 12.2 microM, respectively. In this study, the antituberculosis activity of compound 3 was further investigated. Its activity appeared to be very specific for organisms of the M. tuberculosis complex and it effected significant reductions of bacterial numbers in infected macrophages with an EC(90) of 4.1 microM. More importantly, the increased in vitro antituberculosis activity of the corresponding acid (compound 4) at pH 6.0 suggested that it may be active in vivo in an acidic environment produced as a consequence of inflammation in the lungs of TB patients. The fact that various ester bioisosteres of compound 3 lost anti-TB activity further suggested that the ester compound 3 may function as a prodrug. The detailed structure-activity relationships (SARs) from this study should facilitate our ultimate goal of improving the anti-TB potency of this isoxazole ester series. PMID- 20022501 TI - Synthesis of 4'-ester analogs of resveratrol and their evaluation in malignant melanoma and pancreatic cell lines. AB - 4'-Ester analogs of the disease preventative agent resveratrol were synthesized and evaluated for their potential as anti-melanoma and pancreatic cancer agents. A decarbonylative Heck coupling was used to assemble the protected stilbene core structure. The 4'-acetate and the palmitoate analogs demonstrated selective activity with DM443 and DM738 cells over normal NHDF cells. PMID- 20022502 TI - Chaperone-like activity of the acute-phase component human serum alpha 1-acid glycoprotein: inhibition of thermal- and chemical-induced aggregation of various proteins. AB - In vitro chaperone-like activity of the acute-phase component and plasma drug transporter human alpha(1)-acid glycoprotein (AAG) has been shown for the first time. AAG suppressed thermal aggregation of a variety of unrelated enzymatic (e.g., aldolase, catalase, enolase, carbonic anhydrase) and non-enzymatic proteins (beta-lactoglobulin, ovotransferrin) and it also prevented dithiothreitol induced aggregation of insulin. The anti-aggregation ability of AAG was abolished/reduced upon drug binding suggesting that protein-protein interactions established between the lipocalin beta-barrel fold of AAG and hydrophobic surfaces of the stressed proteins are involved in the chaperone-like activity. The results shed some light on the possible biological function of this enigmatic protein and suggest that besides haptoglobin, clusterin, fibrinogen and alpha(2)-macroglobulin AAG can be considered as a novel member of the extracellular molecular chaperones found in human body fluids. PMID- 20022503 TI - Synthesis and antidepressant activity of optical isomers of 2-(4-benzylpiperazin 1-yl)-1-(5-chloro-6-methoxynaphthalen-2-yl) propan-1-ol (SIPI5056). AB - Four optical isomers of SIPI5056 were synthesized and evaluated for their antidepressant activities and acute toxicities as novel multiple reuptake inhibitors of monoamine transmitters. Chiral alanines were used as educts to prepare their respective target compounds in nine steps. Pharmacological results showed that the (1R,2S)-SIPI5056 isomer has higher inhibitory activity and lower toxicity than other three isomers and is worthy of further development. PMID- 20022504 TI - Synthesis and biological evaluation of new potential inhibitors of N acylethanolamine hydrolyzing acid amidase. AB - N-Acylethanolamines, including N-palmitoyl-ethanolamine (PEA), are hydrolyzed to the corresponding fatty acids and ethanolamine by fatty acid amide hydrolase (FAAH). Recently, N-acylethanolamine-hydrolyzing acid amidase (NAAA) was identified as being able to specifically hydrolyze PEA. In order to find selective and effective inhibitors of this enzyme, we synthesized and screened several amides, retroamides, esters, retroesters and carbamates of palmitic acid (1-21) and esters with C15 and C17 alkyl chains (22-27). Cyclopentylhexadecanoate (13) exhibited the highest inhibitory activity on NAAA (IC(50)=10.0 microM), without inhibiting FAAH up to 50 microM. Compound 13 may become a useful template to design new NAAA inhibitors. PMID- 20022505 TI - Synthesis and structure-activity relationships of novel furazan-3,4-diamide analogs as potent anti-cancer agents. AB - This study describes the synthesis and structure-activity relationships of a series of furazan-3,4-diamide analogs. 1,2,5-Oxadiazole ring and electron withdrawing substituent on the phenyl ring are proposed to be the important elements which contribute to a significant extent maximal potency of anti proliferation effect. PMID- 20022506 TI - Gomphostenins: two new antimalarial compounds from the leaves of Gomphostemma niveum. AB - Phytochemical investigation of CHCl(3) extract of the Gomphostemma niveum leaves led to the isolation of two new diterpene, compound 1 and 2. Their structures were elucidated by spectroscopic procedures and single crystal XRD. Compound 1 named as Gomphostenin 1 and structure was established as 8-ethyl (5H-furan-2-one, 14-hydroxy, 2-oxo 3, 20 Z(17) diene clerodane, while compound 2 named as Gomphostenin-A; was found to be the acetyl derivative of compound 1 and revealed as 8-ethyl (5H-furan-2-one, 14-acetoxy, 2-oxo 3, 20 Z(17) diene clerodane. In vitro antimalarial activity against Plasmodium falciparum showed that compound 2 was more active than compound 1 and CHCl(3) extract as well; with IC(50) value of 3.4 microg/mL. PMID- 20022507 TI - Design and preliminary structure-activity relationship of redox-silent semisynthetic tocotrienol analogues as inhibitors for breast cancer proliferation and invasion. AB - Vitamin E (VE) is a generic term that represents a family of compounds composed of various tocopherol and tocotrienol isoforms. Tocotrienols display potent anti angiogenic and antiproliferative activities. Redox-silent tocotrienol analogues also display potent anticancer activity. The ultimate objective of this study was to develop semisynthetically C-6-modified redox-silent tocotrienol analogues with enhanced antiproliferative and anti-invasive activities as compared to their parent compound. Examples of these are carbamate and ether analogues of alpha-, gamma-, and delta-tocotrienols (1-3). Various aliphatic, olefinic, and aromatic substituents were used. Steric limitation, electrostatic, hydrogen bond donor (HBD) and hydrogen bond acceptor (HBA) properties were varied at this position and the biological activities of these derivatives were tested. Three-dimensional quantitative structure-activity relationship (3D QSAR) studies were performed using Comparative Molecular Field (CoMFA) and Comparative Molecular Similarity Indices Analyses (CoMSIA) to better understand the structural basis for biological activity and guide the future design of more potent VE analogues. PMID- 20022508 TI - Cell permeability of Py-Im-polyamide-fluorescein conjugates: Influence of molecular size and Py/Im content. AB - In order to investigate the influence of molecular size and pyrrole (Py)/imidazole (Im) content on the cell permeability of Py-Im-polyamide fluorescein conjugates we systematically designed the Py-polyamides and Im polyamides. Flow cytometric analysis revealed that Py-polyamides, even those with large molecular size, P-15 and P-18, showed good cellular uptake, but Im polyamides showed very poor uptake. Fluorescence microscopy revealed that conjugate P-6 exhibited nuclear localization, while P-18 showed less nuclear stain but intracellular localization, suggesting that increased molecular size is one of the determinants in reducing nuclear access. Furthermore, results for hairpin polyamide conjugates H-1, H-2, and H-3 containing different Py/Im content indicated that cellular uptake increases as the Im residue is reduced. It appears that Py-Im-polyamide has general properties regardless of whether they have a linear or a hairpin structure. PMID- 20022509 TI - Phenolics from Glycyrrhiza glabra roots and their PPAR-gamma ligand-binding activity. AB - Bioassay-guided fractionation of the EtOH extract of licorice (Glycyrrhiza glabra roots), using a GAL-4-PPAR-gamma chimera assay method, resulted in the isolation of 39 phenolics, including 10 new compounds (1-10). The structures of the new compounds were determined by analysis of their spectroscopic data. Among the isolated compounds, 5'-formylglabridin (5), (2R,3R)-3,4',7-trihydroxy-3' prenylflavane (7), echinatin, (3R)-2',3',7-trihydroxy-4'-methoxyisoflavan, kanzonol X, kanzonol W, shinpterocarpin, licoflavanone A, glabrol, shinflavanone, gancaonin L, and glabrone all exhibited significant PPAR-gamma ligand-binding activity. The activity of these compounds at a sample concentration of 10microg/mL was three times more potent than that of 0.5microM troglitazone. PMID- 20022510 TI - Identification and characterisation of 2-aminopyridine inhibitors of checkpoint kinase 2. AB - 5-(Hetero)aryl-3-(4-carboxamidophenyl)-2-aminopyridine inhibitors of CHK2 were identified from high throughput screening of a kinase-focussed compound library. Rapid exploration of the hits through straightforward chemistry established structure-activity relationships and a proposed ATP-competitive binding mode which was verified by X-ray crystallography of several analogues bound to CHK2. Variation of the 5-(hetero)aryl substituent identified bicyclic dioxolane and dioxane groups which improved the affinity and the selectivity of the compounds for CHK2 versus CHK1. The 3-(4-carboxamidophenyl) substituent could be successfully replaced by acyclic omega-aminoalkylamides, which made additional polar interactions within the binding site and led to more potent inhibitors of CHK2. Compounds from this series showed activity in cell-based mechanistic assays for inhibition of CHK2. PMID- 20022511 TI - A new strategy of endosymbiont midgut bacteria in ant (Ponerinae). AB - The location and ultrastructure of bacteria associated with microvilli in the midgut of Odontomachus bauri were examined by transmission electron microscopy. These filamentous type bacteria are the second morphotype described in the midgut of this ant. They colonizes only the ectoperitrophic space, more specifically attaching along microvilli. A thick capsule attaches bacteria to microvilli and protect them from acidic pH and digestive enzymes. Details of the location and association with microvilli are discussed. PMID- 20022512 TI - Measurement of beta-plus emitters by gamma-ray spectrometry. AB - The activity measurement of beta-plus emitters by gamma-ray spectrometry is studied. Experimental measurements are performed with (22)Na, (65)Zn and (64)Cu with sources included in a lead container. For these nuclides, the activity can be derived both from one photon emission peak and from the 511 keV annihilation peak, including annihilation in-flight correction and geometry correction computed by Monte Carlo simulation. The activity values obtained using the two types of peaks show satisfying agreement. The extension of the method to volume sources is discussed. PMID- 20022513 TI - Temperature dependence of gain shift in gamma-ray spectrometry system involving a long connecting cable. AB - This work demonstrates the possibility of a gain shift or a peak-shape deterioration with changes in the temperature of a long connecting cable between a pre-amplifier and a main amplifier in a Ge gamma-ray spectrometry system. The tests were performed for 50m-long RG174/U and RG58C/U cables with and without the termination. Such a temperature effect may cause from the temperature dependence of the resistivity of a central copper wire. In order to minimize such temperature effects in a Ge gamma-ray spectrometry, use of a terminator should be avoided. PMID- 20022514 TI - "Label and go"--a fast and easy radiolabelling method for pellets. AB - For the development and process optimization of pharmaceutical equipment, it is important to investigate the underlying processes. Taking the fluidized bed technology as an example, the study of particle flow pattern and convection of the particles within the functional unity is essential for construction and process improvement. With positron emission particle tracking (PEPT) it is possible to study the real-time particle motion with radiolabelled particles. We established a fast and simple labelling technique with [(18)F]fluoride for pellets composed of Avicel and anion exchange resin. The uptake of activity ranged from 1.3% to 1.7% per mg and 8.6% to 16.3% per pellet. A specific binding of [(18)F]fluoride with increasing degree of anion exchange resin in the pellets could be observed. PMID- 20022515 TI - Study of excitation functions of alpha-particle induced nuclear reactions on holmium for 167Tm production. AB - (167)Tm is a candidate radioisotope for both nuclear medicine diagnostics and therapy due to its emitted Auger-electrons, low energy X- and gamma-rays. In the frame of a systematic study of excitation functions for production of medically relevant radioisotopes by charged particle induced reactions on rare earths, the (165)Ho(alpha,2n)(167)Tm reaction and the (165)Ho(alpha,n)(168)Tm, (165)Ho(alpha,3n)(166)Tm, (165)Ho(alpha,4n)(165)Tm side reactions were measured up to 40 MeV by the stacked foil irradiation technique and gamma-ray spectroscopy. The measured results were compared to the ALICE-IPPE and EMPIRE-II theoretical curves. Thick target yields, impurity levels and specific activities were deduced and compared with the same parameters for other charged particle production routes of (167)Tm. PMID- 20022516 TI - Surgical treatment of synchronous multiple lung cancer located in a different lobe or lung: high survival in node-negative subgroup. AB - BACKGROUND: The International Association for Study of Lung Cancer Staging Committee proposes for the next revision of TNM (tumour, nodes, metastases) classification that additional nodules in a different lobe of the ipsilateral lung moves from an M1 designation to T4, while additional nodule(s) in the contralateral lung should be classified as M1a, because of poorer survival. We analysed the survival after surgery of patients presenting with synchronous lung cancers located in a different lobe or lung. METHODS: A database of 1551 patients operated on for non-small-cell lung cancer (NSCLC) between 1990 and 2007 was evaluated for unilateral (other lobe) (n=15) and bilateral (n=28) synchronous multiple lung cancers. The relationships among the location of tumours, histology, date of surgery (before and after 2000), lymph node metastasis, type of surgery, adjuvant therapy and survival were analysed. RESULTS: The 5-year survival for all synchronous multiple lung cancers (n=43) was 34%, with a median survival of 32 months. Postoperative mortality was 7%. On univariate analysis, only lymph node metastasis and surgery before the year 2000 affected the overall survival adversely, and both prognostic factors maintained a statistical significance on multivariate analysis. The 5-year survivals were 57% and 0% for patients without (n=25) and with (n=18) lymph node metastasis, respectively (p=0.004), and were 43% and 18% for patients operated upon after (n=27) and before (n=16) the year 2000, respectively (p=0.01), perhaps reflecting a better selection process related to the extensive use of positron emission tomography (PET) scanning. The 5-year survival was not different between bilateral (43%) and unilateral (27%) synchronous lung cancers (p=n.s.). CONCLUSIONS: Our data support complete surgical resection of synchronous multiple lung cancers in patients with node-negative NSCLC. Even patients with bilateral lung cancer should not be treated as metastatic disease. Provided there is no evidence of node and distant metastasis, after an extensive preoperative work-up, including PET scanning and mediastinoscopy, bilateral surgical resection should be performed in fit patients. PMID- 20022517 TI - Feasibility of transtracheal surgical lung biopsy in a canine animal model. AB - Successful natural orifice transluminal endoscopic surgery (NOTES) procedures have been reported in animal studies. However, very little is known about the optimal approach for the application of these surgeries in the thoracic cavity. This study presents the feasibility of transtracheal evaluation of pleural cavity in a canine model using the NOTES technique. PMID- 20022518 TI - Lung sealing using the tissue-welding technology in spontaneous pneumothorax. AB - OBJECTIVE: Welding of lung tissue is a new radio-frequency surgical method that allows sealing pulmonary tissue without overheating and damaging the tissue. The objective of the research was to study the results of sealing the lung tissue in a non-resectional procedure for spontaneous pneumothorax comprising ablation of bullae via video-assisted thoracoscopic surgery (VATS). METHODS: We present a series of 133 consecutive patients with primary spontaneous pneumothorax, who were operated on during the past 3 years. Among 133 patients, 123 were men and 10 were women, with an average age of 26 years (from 14 to 59 years). Indications for surgery were pneumothorax recurrence (59 patients), contralateral occurrence (13), bilateral pneumothorax (one) and haemopneumothorax (two). Prolonged air leakage for more than 2 days was observed in 58 patients. We used the tissue welding technology and an original bipolar hand-piece for bullae electroablation and lung sealing. Conventional apical pleural abrasion was carried out in all cases. Chest tubes were removed 48 h postoperatively by protocol. RESULTS: Intra operatively, emphysema-like changes and blebs under 1cm were seen in 29 patients (22%) and bullae of 1-2 cm in 48 patients (36%); in 56 cases (42%) the size of bullae exceeded 2 cm. In all cases, lung sealing was achieved by tissue welding alone, without using staplers, sutures, glues and sealants. The operating time depended on the presence of adhesions and the number of bullae, but did not exceed 65 min. Postoperative air leakage for 1-6 days was observed in six patients. Neither mortality nor major morbidity was observed. There were seven recurrences (5.2%). CONCLUSIONS: The tissue-welding procedure is easy to perform through VATS and is efficient for ablation of bullae of any size. Leak-proof sealing is achieved, allowing us to repair the pulmonary-pleural fistula, thus being a non-resectional alternative to wedge resection. No conventional wound closing devices are needed. PMID- 20022519 TI - Pulmonary function after pulmonary resection by posterior thoracotomy, anterior thoracotomy or video-assisted surgery. AB - OBJECTIVE: Predicted postoperative forced expiratory volume in 1s (ppoFEV1) is estimated in lung cancer patients before pulmonary resection, as well as the clinical stage. This study aims to evaluate ppoFEV1 and ppo-vital capacity (ppoVC) on postoperative day 7 (POD7) and to compare the results following video assisted thoracic surgery (VATS) and open thoracotomy procedures. METHODS: Of the 155 patients who underwent pulmonary resection, 70 had VATS; 30 had muscle sparing thoracotomy (anterior limiting thoracotomy (AL)); and 55 had postero lateral thoracotomy (PL). VC and FEV1 were measured on POD7 and compared with the VC and FEV1 before surgery using analysis of covariance (ANCOVA). The ratio of the actual- and the ppoVC and FEV1 was evaluated to identify factors associated with variations in postoperative residual VC/FEV1. RESULTS: There were significant differences by analysis of covariance (ANCOVA) in the VC/FEV1 among the three surgical approaches. In the VATS group, the VC ratio and the FEV1 ratio were 96.5% and 94.7%, respectively; they were significantly higher in the VATS group than in the thoracotomy group (AL: 90.4% and 90.1%, respectively; PL: 87.4% and 87.6%, respectively). Non-chronic obstructive pulmonary disease (COPD) and upper lobectomy were also associated with a low VC ratio and FEV1 ratio. CONCLUSION: Predicted postoperative pulmonary function might be overestimated in COPD patients or in those undergoing VATS or lower lobectomy. PMID- 20022520 TI - Giant right coronary pseudo-aneurysm 6 years after angioplasty and stenting. PMID- 20022521 TI - Pseudo-aneurysm of the left carotid artery after surgery for aortic arch interruption. PMID- 20022522 TI - From thorax to the abdomen--an unusual course of a post-infarction left ventricular pseudo-aneurysm. PMID- 20022523 TI - Health economics of blood transfusion safety--focus on sub-Saharan Africa. AB - BACKGROUND AND OBJECTIVES: Health economics provides a standardised methodology for valid comparisons of interventions in different fields of health care. This review discusses the health economic evaluations of strategies to enhance blood product safety in sub-Saharan Africa. METHODS: We reviewed health economic methodology with special reference to cost-effectiveness analysis. We searched the literature for cost-effectiveness in blood product safety in sub-Saharan Africa. RESULT: HIV-antibody screening in different settings in sub-Saharan Africa showed health gains and saved costs. Except for adding HIV-p24 screening, adding other tests such as nucleic acid amplification testing (NAT) to HIV antibody screening displayed incremental cost-effectiveness ratios greater than the WHO/World Bank specified threshold for cost-effectiveness. The addition of HIV-p24 in combination with HCV antibody/antigen screening and multiplex (HBV, HCV and HIV) NAT in pools of 24 may also be cost-effective options for Ghana. CONCLUSIONS: From a health economic viewpoint, HIV-antibody screening should always be implemented in sub-Saharan Africa. The addition of HIV-p24 antigen screening, in combination with HCV antibody/antigen screening and multiplex (HBV, HCV and HIV) NAT in pools of 24 may be feasible options for Ghana. Suggestions for future health economic evaluations of blood transfusion safety interventions in sub-Saharan Africa are: mis-transfusion, laboratory quality and donor management. PMID- 20022524 TI - Percutaneous distraction osteogenesis for treatment of brachymetatarsia. AB - Brachymetatarsia is not an unusual deformity and is often associated with functional and cosmetic issues that warrant surgical reconstruction. Lengthening of the affected metatarsal can be undertaken on an acute basis in a single operative procedure that involves the use of a bone graft or by means of gradual callus distraction. Because of the risk of metatarsophalangeal joint malalignment, it is important for the surgeon to take steps to stabilize the joint during the lengthening process. In this report, a percutaneous method of callus distraction for repair of brachymetatarsia and maintenance of metatarsophalangeal joint alignment is presented. PMID- 20022525 TI - Irreducible fracture dislocation of the ankle caused by tibialis posterior tendon interposition. AB - A patient with severe irreducible open fracture dislocation of the ankle was admitted to our emergency department. After wound irrigation and debridement, skeletal traction was applied to the calcaneus to minimize soft tissue injury and swelling. The patient was followed in traction for 1 week, after which reduction and fixation of the fibula was attempted but not achieved. We extended the incision distally, visualized the ankle, and located the tibialis posterior tendon between the distal tibia and fibula, thereby inhibiting the reduction. The tendon coursed into the tibiotalar joint anteriorly and pushed the talus anterolaterally. After manipulation of the tendon to its anatomically correct location, the ankle was easily reduced. The wound at the medial side was closed with a fasciocutaneous rotational flap. The ankle was then immobilized for 6 weeks postoperatively. The patient regained her full range of motion, and there were no problems with the tibialis posterior tendon, such as rupture or insufficiency. Isolated tibialis posterior tendon interposition between the distal tibiofibular and tibiotalar joints has rarely been reported, and can inhibit anatomical reduction of the fractured ankle. PMID- 20022526 TI - The porcine small intestinal submucosa (SIS) patch in foot and ankle reconstruction. AB - We undertook a retrospective cohort study of 54 patients who underwent foot and ankle soft tissue reconstructive surgery augmented with a porcine small intestinal submucosal (SIS) patch. The mean patient age was 44 (range 17 to 68) years, there were 21 (38.89%) males in the cohort, and the mean follow-up duration was 1080 (range 365 to 1943) days. Clinical outcomes were considered excellent in 46 (85.19%) patients, good/fair in 3 (5.56%) patients, and poor in 5 (9.26%) patients; and no adverse events attributable to the xenograft were observed. Direct SIS patch failure, resulting in stretching of the repair, re tear, or tendon stenosis, occurred in 3 (5.56%) patients, and delayed incision healing occurred in 6 (11.11%) patients. Based on our observations, we concluded that the porcine SIS xenograft, when used to augment cellular and vascular in growth, is a viable adjunct to musculoskeletal reconstructions of the foot and ankle. PMID- 20022527 TI - Inflammatory reaction to implanted equine pericardium xenograft. AB - Equine pericardium xenograft is now widely used to augment Achilles tendon repair. In this article, we describe the case of a postoperative complication in a healthy 37-year-old male who underwent primary repair, augmented with equine pericardium, of his acutely ruptured Achilles tendon. At 4 months postoperative, an indurated and fluctuant subcutaneous soft tissue reaction was noted at the repair site. Further questioning led the patient to recall having an allergy to horses, although it had been a very long time since he had experienced any symptoms related to the allergy and, as such, he failed to mention this important fact at the time of his initial examination. At 11 months postoperative, the xenograft was removed and, thereafter, the wound healed uneventfully. At 6 months following removal of the xenograft, the patient displayed no evidence of allergy or Achilles tendon dysfunction. PMID- 20022528 TI - Generation and characterization of human delta-globin-specific monoclonal antibodies. AB - The human delta-globin chain, unique to the hemoglobin A2 (HbA2) heterotetramer, is important for the evaluation of hemoglobinopathy. However, there are no well defined antibodies specific for the delta-globin chain, a fact that is attributed a striking similarity (93%) in amino acid sequence between delta-globin and beta globin of the hemoglobin A (HbA). In this study, two monoclonal antibodies (mAbs) against the delta-globin chain were generated and designated as 2H4 and 1H11. These antibodies were specific to HbA2 and do not cross-react with HbA and HbF (fetal hemoglobin). Moreover, the expression of HbA2 in fetal liver and mature erythrocytes was determined using these two mAbs. In addition to being useful tools for research or diagnosis, these antibodies could be valuable for development of rapid and effective antibody-based immunoassays of HbA2 expression in erythroid cells and non-erythroid tissue. PMID- 20022529 TI - Risk factors for calving assistance and dystocia in pasture-based Holstein Friesian heifers and cows in Ireland. AB - The incidence of and risk factors associated with calving assistance and dystocia in pasture-based dairy herds were determined from analysis of 152,641 records of full-term calvings from Holstein-Friesian dams served by artificial insemination (AI) sires of seven breeds in herds of 20 calvings or more per year over 4 years. The overall incidence of calving assistance and dystocia was 31.1% and 6.8%, respectively. The incidence in primiparae and pluriparae was 40.0% and 28.2% for assistance, and 9.3% and 5.8% for dystocia, respectively. Association analyses were undertaken using generalised estimating equations using a logit link function. The likelihood of calving assistance or dystocia did not change over time but was greatest in autumn and in spring. The likelihood of calving assistance or dystocia was greater for males in primiparae and males sired by Charolais sires. The odds of calving assistance and dystocia were greater in twin calves (OR 2.0 and 2.4; P<0.001) and in dams that had dystocia at the previous calving (OR 1.65 and 2.9; P<0.001). The logit of the probability of calving assistance and dystocia increased linearly per unit increase in sire predicted transmitting ability for direct calving difficulty. The probability of assisted calving, but not dystocia, increased linearly in primiparae as animals calved at a younger age relative to the median age at first calving. Herd size and day of the week of calving were not associated with the odds of dystocia. Stakeholders must focus on identified modifiable risk factors to control the incidence of dystocia in dairy herds. PMID- 20022530 TI - Pyruvate dehydrogenase E2 deficiency: a potentially treatable cause of episodic dystonia. AB - The association of progressive episodic dystonia and learning disability with distinctive neuroimaging findings may lead to consideration of atypical Pantothenate Kinase Associated Neurodegeneration (PKAN) and investigations directed towards that diagnosis. Recent reports indicate that deficiency of dihydrolipoamide acetyltransferase, the E2 component of the pyruvate dehydrogenase complex, may present similarly, and that this disorder should also be considered in the differential diagnosis. We describe two sisters with early onset episodic dystonia and pyruvate dehydrogenase deficiency caused by defects in the E2 subunit. Both have neuroimaging features similar to previously described patients and have mutations in the DLAT gene. As this condition is potentially treatable with a ketogenic diet, the possibility of this diagnosis should be considered in similar cases. PMID- 20022531 TI - Ames dwarf (Prop1(df)/Prop1(df)) mice display increased sensitivity of the major GH-signaling pathways in liver and skeletal muscle. AB - CONTEXT: Growth hormone (GH) is an anabolic hormone that regulates growth and metabolism. Ames dwarf mice are natural mutants for Prop1, with impaired development of anterior pituitary and undetectable levels of circulating GH, prolactin and TSH. They constitute an endocrine model of life-long GH-deficiency. The main signaling cascades activated by GH binding to its receptor are the JAK2/STATs, PI-3K/Akt and the MAPK Erk1/2 pathways. OBJECTIVES: We have previously reported that GH-induced STAT5 activation was higher in Ames dwarf mice liver compared to non-dwarf controls. The aim of this study was to evaluate the principal components of the main GH-signaling pathways under GH-deficiency in liver and skeletal muscle, another GH-target tissue. METHODS: Ames dwarf mice and their non-dwarf siblings were assessed. Animals were injected i.p. with GH or saline 15min before tissue removal. Protein content and phosphorylation of signaling mediators were determined by immunoblotting of tissue solubilizates. RESULTS: GH was able to induce STAT5 and STAT3 tyrosine phosphorylation in both liver and muscle, but the response was higher for Ames dwarf mice than for non dwarf controls. When Erk1/2 activation was assessed in liver, only dwarf mice showed GH-induced phosphorylation, while in muscle no response to the hormone was found in either genotype. GH-induced Akt phosphorylation at Ser473 in liver was only detected in dwarf mice. In skeletal muscle, both normal and dwarf mice responded to a GH stimulus, although dwarf mice presented higher GH activation levels. The phosphorylation of GSK-3, a substrate of Akt, increased upon hormone stimulation only in dwarf mice in both tissues. In contrast, no differences in the phosphorylation of mTOR, another substrate of Akt, were observed after GH stimulus, either in normal or dwarf mice in liver, while we were unable to determine mTOR in muscle. Protein content of GH-receptor and of the signaling mediators studied did not vary between normal and dwarf animals in the assessed tissues. CONCLUSION: These results show that several components of the main GH signaling pathways exhibit enhanced sensitivity to the hormone in liver and muscle of Ames dwarf mice. PMID- 20022533 TI - Posttraumatic psoriatic osteitis of the frontal bone successfully treated with etanercept. Report of a case. AB - We report a case of aseptic osteomyelitis of the frontal bone that developed after a local injury in an 8-year-old girl with psoriasis of the scalp as a predisposing factor. Follow-up was 36 years (1972 to 2008). Enthesitis of the forehead muscles was a plausible pathophysiological mechanism. The symptoms responded to anti-inflammatory medications and resolved immediately after the introduction of etanercept therapy. The most interesting feature of this case of psoriatic osteomyelitis is the involvement of a skull bone. PMID- 20022534 TI - Socioeconomic impact of ankylosing spondylitis in Tunisia. AB - OBJECTIVE: Ankylosing spondylitis (AS) is the second most common chronic inflammatory joint disease after rheumatoid arthritis and causes substantial functional impairment, two features that generate a heavy socioeconomic burden. Here, our objective was to assess the socioeconomic impact of AS and to identify factors associated with higher costs. PATIENTS AND METHODS: We retrospectively reviewed the medical charts of 50 patients with AS seen at the Monastir Public Health Service Hospital over the 6-month period from March to September 2006. The following were evaluated: direct costs of medical care; indirect costs related to work incapacity; and impact on marital life, offspring, social activities, and activities of daily living. RESULTS: There were 42 men and eight women (male-to female ratio, 5.25) with a mean age of 38.9+/-10.8 years (range, 19-60 years). The median mean direct cost of medical care for AS was 426.072 Tunisian Dinars (TND) (266.295 euro) per year, and the interquartile range (IQR) was 270.468 TND. Of the 34 patients who had paid employment, 12 (35%) were on sick leave. The mean indirect cost was 447.4+/-294.3 TND (279.625+/-183.937 euro) per patient per year. The median mean total cost was 873.472 TND (545,92 euro) per patient per year with an IQR of 292,324 TND. Factors associated with higher costs were the use of nonsteroidal anti-inflammatory drugs and higher values of the BASDAI and BASRI. Among married patients, 44.4% reported sexual problems, which correlated with the BASMI; and 37% reported a negative reaction on the part of the healthy spouse. Adverse effects on schooling and quality of life of the children were noted in 29.6% of cases. Among single patients, 30.4% felt their disease was responsible for their unmarried status. The disease adversely affected the ability to carry out many activities of daily living (grooming in 38% of cases, housework in 76%, shopping in 92%, sporting activities in 96%, socializing in 68%, and traveling in 80%). The patients usually reported receiving support from their family, which was physical in 74% of cases, financial in 52%, and psychological in 90%. CONCLUSION: Our data indicate that AS generates a major socioeconomic burden. Most of the factors associated with higher costs were related to greater disease activity. Therefore, early appropriate treatment is crucial. Despite the many socioeconomic problems generated by AS, the patients remained connected to their social network thanks to support from their family and friends. PMID- 20022535 TI - Extraspinal sciatica revealing late metastatic disease from parotid carcinoma. AB - Sciatica is a clinical symptom usually caused by a disk herniation and less often by other conditions such as tumors, infections, or inflammatory diseases. We report the case of a woman in whom sciatica led to the identification of a large pelvic metastasis from a carcinoma of the parotid gland. PMID- 20022536 TI - Hematogenous Staphylococcus aureus discitis in adults can start outside the vertebral body. AB - BACKGROUND: Textbooks describe hematogenous discitis in adults as starting in the anterior vertebral body near the endplates. CASE REPORT: Hematogenous Staphylococcus aureus discitis developed at L1-L2 in an 81-year-old woman. On the first magnetic resonance imaging (MRI) scan performed 4 days after symptom onset, the only abnormalities were high signal from the disk on T2-weighted images, gadolinium enhancement of the soft tissues anterior to L1-L2, and epidural involvement. The endplates and vertebral bodies were normal. A subsequent MRI scan showed features typical of infectious discitis. DISCUSSION: We found a single similar case report in the medical literature, indicating that discitis starting outside the anterior vertebral body is rare. Nevertheless, an increasing number of similar cases may be diagnosed in the future, as MRI is being performed increasingly early in patients with a clinical suspicion of infectious discitis. CONCLUSION: Hematogenous infectious discitis in adults may start in some patients within the disk, soft tissues anterior to the disk, or epidural space. To avoid diagnostic and therapeutic delays, physicians should be aware of this unusual presentation of infectious discitis on very early MRI scans. When the clinical picture suggests discitis and findings from the early MRI scan are atypical, a repeat MRI scan should be obtained 1 week later to confirm the diagnosis. PMID- 20022537 TI - Correspondence to the article by Park Min-Chan "Pro-inflammatory effect of leptin on peripheral blood mononuclear cells of patients with ankylosing spondylitis" Joint Bone Spine 2009;76:170-75. PMID- 20022538 TI - New insights in synovial angiogenesis. AB - Angiogenesis is the formation of new capillaries from pre-existing vessels. A number of soluble and cell-bound factors may stimulate neovascularization. The perpetuation of angiogenesis involving numerous soluble and cell surface-bound mediators has been associated with rheumatoid arthritis (RA). These angiogenic mediators, among others, include growth factors, primarily vascular endothelial growth factor (VEGF) and hypoxia-inducible factors (HIFs), as well as pro inflammatory cytokines, various chemokines, cell adhesion molecules, proteases and others. Among the several potential angiogenesis inhibitors, targeting of VEGF, HIF-1, angiopoietin and the alpha(V)beta(3) integrin, as well as some endogenous or synthetic compounds including angiostatin, endostatin, paclitaxel, fumagillin analogues, 2-methoxyestradiol and thalidomide seems to be promising for the management of synovial inflammation and angiogenesis. A complete review of antiangiogenic drugs used in animal models of arthritis or human RA is available in a table. PMID- 20022539 TI - Preventing the gastrointestinal adverse effects of nonsteroidal anti-inflammatory drugs: from risk factor identification to risk factor intervention. AB - Nonsteroidal anti-inflammatory drugs (NSAIDs) have huge prescription volumes, for two main reasons: the aging of the population is increasing the prevalence of diseases that respond to NSAIDs, such as osteoarthritis; and NSAIDs are highly effective drugs that contribute crucially to the management of many diseases. In France, the number of physician orders that include an NSAID is estimated at 25 to 30 million per year. Nevertheless, the use of NSAIDs is limited by adverse effects. The gastrointestinal tract is the main target of NSAID toxicity, and NSAID therapy is among the leading causes of bleeding from upper gastrointestinal ulcers. Adverse events targeting the lower gastrointestinal tract are also of concern, although they receive less attention. To effectively prevent NSAID toxicity, it must be recognized that the risk of adverse events can be diminished but not eliminated. Therefore, the risk/benefit ratio must be carefully evaluated at each prescription. A number of risk factors should be emphasized. Thus, the risk increases with age, and there is a sharp risk increase at 60 years of age. Other risk factors include a history of ulcers (most notably with bleeding), the use of high NSAID dosages, Helicobacter pylori infection, and the concomitant use of antiplatelet agents. Minimizing NSAID-related gastrointestinal toxicity requires a careful risk factor evaluation; selection of the most appropriate NSAID and NSAID dosage; and, in some patients, prophylactic gastroprotective therapy, for instance with a proton pump inhibitor. Gastrointestinal symptoms either have no value for predicting gastrointestinal events or occur too late to serve as alarm signals. The toxicity advantages of cyclooxygenase-2 inhibitors seem modest and do not eliminate the need for this rational prescription strategy. PMID- 20022540 TI - [New rules for prenatal screening for T21: collateral damages]. PMID- 20022541 TI - [A response by E. Phan to the article by B. Langer et al.: Low-risk pregnancy and low-risk delivery: can we really consider it?]]. PMID- 20022542 TI - [HBsAg screening during pregnancy in the French province Picardy]. AB - OBJECTIVE: Screening for maternal hepatitis B surface antigen (HBsAg) is mandatory in France since 1992, however, no evaluation is available. We studied the traceability of HBsAg screening and its prevalence in pregnant women in Picardy for year 2006. PATIENTS AND METHODS: Traceability of HBsAg screening was studied in a sample of 1198 hospital case files, which were randomized and stratified for all the 20 clinics of the region (22,114 deliveries), both public and private. HBsAg prevalence was also studied using various registries (PMSI national database of medical acts performed during hospitalization, central pharmacies and obstetric theatres). RESULTS: The traceability of the screening was lacking in 9.9% (range: 0-34.7%, depending on the maternity clinic). The prevalence of HBsAg during pregnancy was 1.8 per 1000 women (upper limit: 4.3 per 1000) from the case files sample. Registries examination showed large variations of HBsAg's prevalence from 0 to 12.0 per 1000 (mean: 2.9; CI 95%: 7 to 17) among the region. DISCUSSION AND CONCLUSION: HBsAg traceability during pregnancy must be improved. HBsAg prevalence largely varies among maternity clinics and is a significant issue which is underestimated in France. PMID- 20022543 TI - [Egg donation: regulation of the donation and the hidden face of the cross-border reproductive care]. AB - In France egg donation is not sufficient to meet the needs of infertile couples. As a consequence there is a long waiting time for those couples who may be driven to search solutions abroad. Nonetheless in some countries practice of egg donation raises medical and ethical questions regarding the quality of care provided to the recipients as well as the recruitment and follow-up of egg donors. Meanwhile the scope of this type of "cross-border reproductive care" remains difficult to assess, even though this trend seems to be growing and is reinforced by a recent regulation facilitating the free movement of people seeking cross-border health care within the European Union. These observations lead to question the possible causes of "cross-border reproductive care" in the light of national laws, to identify potential consequences in order to suggest possible actions at both national and international level. PMID- 20022544 TI - Sonoelectrochemical treatment of water polluted with trichloroacetic acid: from sonovoltammetry to pre-pilot plant scale. AB - The sonoelectrochemical treatment of aqueous solutions of trichloroacetic acid (TCAA) has been scaled-up from the voltammetric analysis to pre-pilot stage. The degradation in absence of ultrasound field has yield to a poor performance which has been improved in presence of ultrasound. The sonovoltametry study has provided the range of potentials and/or current densities to be used with the lowest current efficiency penalty. Sonoelectrolyses at batch scale (carried out with a horn-transducer 24 kHz positioned at about 3 cm from the surface of the electrode) achieved little improvement in the degradation. However, when a specifically designed sonoelectrochemical reactor (not optimized) was used during the scale-up, the presence of ultrasound field provided better results (fractional conversion 97%, degradation efficiency 26%, selectivity 0.92 and current efficiency 8%) at lower ultrasonic intensities and volumetric flow. PMID- 20022545 TI - Power ultrasonic transducers with extensive radiators for industrial processing. AB - High-power ultrasonics (HPU) is a green emerging technology that offers a great potential for a wide range of industrial processes. Nevertheless such potential have remained restricted during many years to a limited number of applications which reached commercial development. The possible major problem for extending the range of HPU industrial applications has been the lack of power ultrasonic transducers for large-scale application, adapted to the requirements of each specific problem with high efficiency and power capacity. A new family of HPU transducers with extensive radiators have been recently introduced. It comprises a variety of transducer types designed with the radiators adapted to different specific uses in fluids and multi-phase media. Such transducers implement high power capacity, high efficiency and radiation pattern control. In addition, their design incorporate strategies to eliminate or reduce modal interactions produced at high power as a consequence of their nonlinear behaviour. The introduction of such new transducers has significantly contributed to the development at semi industrial and industrial level of a number of processes in the food and beverage industry, in environment and in manufacturing. This paper deals with the basic structure and main characteristics of such transducers as well as their performance in the developed application processes. PMID- 20022546 TI - Ultrasound-assisted oxidative desulfurization of liquid fuels and its industrial application. AB - Latest environmental regulations require a very deep desulfurization to meet the ultra-low sulfur diesel (ULSD, 15 ppm sulfur) specifications. Due to the disadvantages of hydrotreating technology on the slashing production conditions, costs and safety as well as environmental protection, the ultrasound-assisted oxidative desulfurization (UAOD) as an alternative technology has been developed. UAOD process selectively oxidizes sulfur in common thiophenes in diesel to sulfoxides and sulfones which can be removed via selective adsorption or extractant. SulphCo has successfully used a 5000 barrel/day mobile "Sonocracking" unit to duplicate on a commercial scale its proprietary process that applies ultrasonics at relatively low temperatures and pressures. The UAOD technology estimate capital costs less than half the cost of a new high-pressure hydrotreater. The physical and chemical mechanisms of UAOD process are illustrated, and the effective factors, such as ultrasonic frequency and power, oxidants, catalysts, phase-transfer agent, extractant and adsorbent, on reaction kinetics and product recovery are discussed in this review. PMID- 20022547 TI - Ultrasound-assisted dehydrogenation of 5-acetyl-3,4-dihydropyrimidin-2(1H)-ones. AB - In this study, various 5-acetyl-3,4-dihydropyrimidin-2(1H)-ones were synthesized and the dehydrogenation of these compounds by potassium peroxydisulfate in aqueous acetonitrile under thermal and sono-thermal conditions were investigated. Whereas the effect of the nature of 4-substituent influences the rate of reaction, the application of sonic waves decreases drastically the time of thermal reaction. PMID- 20022548 TI - Potential applications of sonochemistry in spent nuclear fuel reprocessing: a short review. AB - The industrial treatment of spent nuclear fuel is based upon a hydrometallurgical process in nitric acid medium. In order to minimize the volume of radioactive waste it seems interesting to generate the reactive species in situ in such solutions using ultrasonic irradiation without addition of salt-forming reagents. This review summarizes for the first time the versatile sonochemical processes with uranium, neptunium and plutonium in homogeneous nitric acid solutions and heterogeneous systems. The dissolution of refractory solids, ultrasonically driven liquid-liquid extraction and the sonochemical degradation of the volatile products of organic solvent radiolysis issued from PUREX process are considered. Also the guidelines for required further work to ensure successful application of the studied processes at industrial scale are discussed. PMID- 20022549 TI - Social and clinical determinants of quality of life in Parkinson's disease in a Russian cohort study. AB - Parkinson's disease (PD) is a chronic neurodegenerative disorder that has a major impact on health and longevity in Eastern countries. Studies investigating health related quality of life (HRQoL) in Eastern European and Asian countries are scarce. The objective of this cross-sectional survey was to assess HRQoL in Russian patients with PD and identify its social and clinical determinants. The study included 100 outpatients with idiopathic PD and 100 controls. Patients were consecutively recruited from the neurological department of the Russian Medical State University in Moscow between October 2004 and December 2005. Regional healthy controls were matched for age and sex. The evaluation of HRQoL was performed using the EuroQol instrument (EQ-5D and EQ VAS). Disease severity was assessed using the Unified Parkinson's Disease Rating Scale (UPDRS). Multivariate regression analyses were used to identify independent determinants of HRQoL. HRQoL was more notably decreased in PD patients than in controls (98% versus 74% of individuals with moderate or severe problems in at least one dimension of the EQ-5D (p < 0.001), respectively). As compared to patients, the controls reported a higher mean EQ VAS score (74.0 +/- 16.0 versus 47.7 +/- 16.7, p < 0.001). Social and clinical determinants of HRQoL were age, disease severity, dystonia, depression, dementia and social support. While the HRQoL of patients with PD in Western countries is predominately affected by clinical parameters, social factors play an important role in Eastern countries. Our data should be considered in the development of national healthcare programs that seek to provide better social services support for patients with PD. PMID- 20022550 TI - Early experiences matter: lasting effect of concentrated disadvantage on children's language and cognitive outcomes. AB - A small but provocative literature suggests that neighbourhood socioeconomic conditions experienced by children early in life influence a variety of health and developmental outcomes later in life. We contribute to this literature by testing the degree to which neighbourhood socioeconomic conditions that children experience in Kindergarten influence their later language and cognitive outcomes in early adolescence, over and above current neighbourhood context and various child-level covariates including scores on a Kindergarten measure of school readiness. Cross-classified random effects modelling (CCREM) analyses were performed on a study population of 2648 urban children residing throughout the province of British Columbia, Canada, who were followed longitudinally from Kindergarten (age 5/6) to Grade 7 (age 12/13). Findings demonstrate that neighbourhood concentrated disadvantage experienced during Kindergarten has a durable, negative effect on children's reading comprehension outcomes seven years later-providing evidence that early social contextual experiences play a critical role in the lives of children. Possible explanations and future directions are discussed. PMID- 20022551 TI - Health inequalities in urban and rural Canada: comparing inequalities in survival according to an individual and area-based deprivation index. AB - Social health inequalities are smaller in rural than urban areas because, some argue, people's social conditions are estimated at a small-area level, hiding variability at the individual or household level. This paper compares inequalities in survival according to an individual and area-based version of a deprivation index among a large sample of Canadians living in various urban and rural settings. Results show that survival inequalities in small towns and rural areas are lower than elsewhere when an area-based index is used, except in the remote hinterland, but of equal magnitude when the individual version of this index is considered. PMID- 20022552 TI - The reproducibility of a new power mapping instrument based on the phase shifting schlieren method for the measurement of spherical and toric contact lenses. AB - PURPOSE: To assess a new method of power measurement of soft and rigid contact lenses. The method is the phase shifting schlieren method, as embodied in the Nimo TR1504 instrument. MATERIALS AND METHODS: Three Nimo TR1504 instruments were used to measure the power related dimensions of: (a) a range of custom toric rigid lenses; (b) a range of commercially available spherical hydrogel lenses; and (c) a commercially available range of toric silicone hydrogel lenses. The measurements were carried out using a standard ISO ring test protocol where independent tests were carried out under conditions of reproducibility. The analysis of the measurements was carried out using ISO methods which enabled the reproducibility standard deviation, SR, of the method to be calculated. RESULTS: The results show that this new method has SR of 0.048D for spherical soft (hydrogel) lenses. This means the back vertex power of spherical soft lenses having a power in the range +/-10.0D can be determined to current ISO product tolerances with a single measurement. The method has SR of 0.059D for sphere power and 0.093D for cylinder power for toric soft lenses having powers in the range +/-10.0D and cylinder powers in the range +/-2.0D. A single measurement will determine sphere power to current ISO tolerance limits with 95% confidence while two measurements are required to determine the cylinder power to the same confidence level. PMID- 20022553 TI - Chemoprevention of oral cancer in animal models, and effect on leukoplakias in human patients with ZengShengPing, a mixture of medicinal herbs. AB - ZengShengPing (ZSP), a mixture of six medicinal herbs, has been reported to prevent esophageal squamous cell carcinoma (SCC) in human patients with dysplasia. This study was designed to investigate the chemopreventive effects of ZSP on oral cancer in animal models and human patients. In the 7,12 dimethylbenz[a]anthracene (DMBA)-induced hamster cheek pouch model, ZSP (6g/kgBW/day by gavage for 10 weeks) significantly reduced the number of visible tumor, the tumor volume, and the incidence of SCC (P<0.01). Two biomarkers associated with cell proliferation, silver stained nucleolar organizer region (AgNOR) and proliferating cell nuclear antigen (PCNA)-labeling index, were also significantly suppressed by ZSP treatment (P<0.01). In the 4-nitroquinoline 1 oxide (4NQO)-induced oro-esophageal cancer model in mice, ZSP (10% in diet) also significantly reduced the incidence of tongue SCC from 55.2% (16/29) to 22.2% (6/27) (P<0.05), and slightly reduced the incidence of esophageal SCC from 34.5% (10/29) to 22.2% (6/27). Furthermore, in a randomized clinical trial on patients with oral leukoplakia, ZSP (4 tablets, 3 times per day for 8-12months) reduced the size of oral lesion in 67.8% (40/59) patients, whereas the placebo was effective in 17% (9/53) patients (P<0.01). Such an effect was associated with significant decrease of AgNOR and PCNA-labeling index. In summary, our studies have demonstrated the chemopreventive effects of ZSP on two animal models of oral cancer, and human patients with oral leukoplakia. PMID- 20022554 TI - A DFT/TD DFT study of the structure and spectroscopic properties of 5-methyl-2-(8 quinolinyl)benzoxazole and its complexes with Zn(II) ion. AB - The structure and spectroscopic properties of 5-methyl-2-(8 quinolinyl)benzoxazole and its complexes with Zn(II) ion were studied using a DFT and TD DFT methods with def2-TZVP basis set. It was shown that the type of functional used (B3-LYP or pbe0) implemented in TURBOMOLE package does not have essential influence on the geometry (small differences in bond length, valence and dihedral angles) of studied compounds in both ground and excited states. However, significant differences were obtained for the position of vertical absorption and emission transition but not for the oscillator strength of transition. Application of pbe0 functional seems to reproduce better the experimental spectrum. PMID- 20022555 TI - Study of lanthanide aromatic acid complexes in silica gels by photoacoustic spectroscopy. AB - Lanthanide complexes Ln(p-ABA)(3).H(2)O (p-ABA: p-aminobenzoic acid; Ln(3+):La(3+), Tb(3+) and Er(3+)) have been incorporated into silica gels via a sol-gel method. Upon heat treatment at 120 degrees C, photoacoustic (PA) intensity of the ligand increases for Tb(3+), La(3+) and Er(3+) complexes in silica gels, respectively, while this difference cannot be observed for the samples without heat treatment. Different PA intensities of the samples are interpreted by comparison with their luminescence spectra. The nephelauxetic parameters and PA branching vectors of Er(3+) complex in silica gel have been calculated. Spectral results indicate that p-ABA does not coordinate with lanthanide ions in silica matrix without a suitable heat treatment. For the co doped samples, it is shown that the emissions of Tb(3+) are enhanced with addition of La(p-ABA)(3).H(2)O and remarkably quenched with the addition of Er(p ABA)(3).H(2)O. The possible mechanisms for these phenomena are proposed. PMID- 20022556 TI - Sleep-related periodic respiration with central sleep apnea in Leber Hereditary Optic Neuropathy (LHON). PMID- 20022557 TI - Narcolepsy and Behcet's disease: report of a Chinese-Taiwanese case. PMID- 20022558 TI - Recycling and uptake of Si(OH)4 when protozoan grazers feed on diatoms. AB - Herbivory of microzooplankton is an emerging key factor of diatom mortality in the ocean. As part of the microbial loop, protozoan grazers also feed on bacteria that accelerate the degradation of diatom detritus. The potentially pivotal effect of microzooplankton grazing on Si(OH)(4) recycling was investigated with cultures of single-celled diatoms, Thalassiosira pseudonana and Chaetoceros gracilis, and heterotrophic protozoans, the dinoflagellate Oxyrrhis marina and the ciliate Strombidium sp. Both grazers ingested diatoms and the bacteria in the non-axenic cultures. C. gracilis, whose frustule is "armed" with setae, was less suitable as a prey than T. pseudonana. Ingestion rates of T. pseudonana were comparable for O. marina and Strombidium, but the dinoflagellate produced two orders of magnitude more detrital bSiO(2) than the ciliate, due to the higher abundance reached by O. marina. Total net release of Si(OH)(4) was lower in the grazing treatments compared to the control possibly due to the reduced bacterial growth by microzooplankton bacterivory, and to the transient protection of detrital bSiO(2) in discarded feeding vacuoles. Over the first 24h, microzooplankton grazing even led to enhanced uptake of Si(OH)(4) by diatoms, confirming the potential of grazing to influence the silicification of diatom frustules. Subsequently however, the Si dynamics in bottles with grazers turned rapidly from net uptake to net Si(OH)(4) release. Protozoan grazers hence tie Si(OH)(4) recycling into the microbial loop by producing detrital bSiO(2). PMID- 20022560 TI - Living in intermediate hosts: evolutionary adaptations in larval helminths. AB - In the complex life cycles of helminths, life in intermediate hosts poses special problems not covered by standard life history strategy theory. While under selection to reduce mortality and to increase growth, there is the additional problem of transmission between hosts. This review attempts to harmonise classical knowledge of the overall life cycle patterns with recent evolutionary theory as to how larval helminths exploit intermediate host tissues and avoid the gut to maximise fitness in terms of growth and mortality. It also considers the evolutionary rules by which trophically transmitted larvae are expected to increase their transmission rates to the next host. PMID- 20022561 TI - Giardia sex? Yes, but how and how much? AB - Although Giardia is of practical importance as a pathogen and has theoretical importance in evolutionary biology, it is not known whether it ever reproduces sexually. Several recent papers have shed light on this problem, without completely solving it. One paper shows that nuclei in the encysted organism can temporarily fuse and exchange genes; this may explain the genetic similarity of the two nuclei in a cell. Another paper demonstrates that Giardia does undergo sexual reproduction with outcrossing. However, we still do not know whether this involves a meiotic or a parasexual cycle, when it occurs, or how common it is. PMID- 20022559 TI - Hijacking HES1: how tumors co-opt the anti-differentiation strategies of quiescent cells. AB - Quiescent and tumor cells share the ability to evade irreversible cell fates. Recent studies have shown that the transcriptional regulator Hairy and Enhancer of Split 1 (HES1) protects quiescent fibroblasts from differentiation or senescence. HES1 is highly expressed in rhabdomyosarcomas, and the inhibition of HES1 restores differentiation in these cells. Pathways that lead to elevated HES1 levels, such as the Notch and Hedgehog pathways, are frequently upregulated in tumors. Compounds that inhibit these pathways induce differentiation and apoptosis in cancer cells and several are in clinical trials. HES1 might repress gene expression in part by recruiting histone deacetylases (HDACs). HDACs inhibit differentiation, whereas histone deacetylase inhibitors (HDACis) induce differentiation or apoptosis in tumors and are also showing promise as therapeutics. Small molecules that directly target HES1 itself were recently identified. Here, we discuss the importance of HES1 function in quiescent and tumor cells. Elucidating the pathways that control quiescence could provide valuable information not only for treating cancer but also other diseases. PMID- 20022562 TI - Implementation and sustainability of the nursing and midwifery standards for mentoring in the UK. AB - There is growing emphasis on developing sounder processes for ensuring that nursing and midwifery students are appropriately supported and assessed in practice settings, so that they are fit to practice at the point of registration. To support this, the Nursing and Midwifery Council (NMC) in 2006, introduced new mandatory standards for practice education (NMC, 2006). The standards outline mentor responsibility for developing and ensuring the practice competence of students and provide a more defined statement regarding accountability for the decisions that lead to entry to the professional register (NMC, 2006, p. 6). Integral to the standards is the creation of a new role final placement mentors; the 'Sign-off' mentor. The concept of the 'Sign-off' mentor is new and the arrangements for implementing a sustainable approach will be challenging for placement providers. Equally challenging for universities, is the development of a framework that can support, monitor and provide evidence, that regulatory requirements have been met. This paper outlines the complexities associated with maintaining mentor competence, discusses some of the challenges for Sign-off mentors and is intended to contribute to the general discussion about specific aspects of the Nursing and Midwifery Council (NMC) standards and their sustainability in practice. PMID- 20022563 TI - Rising hypospadias rates: disproving a myth. AB - Hypospadias is one of the most common congenital abnormalities occurring in males. In recent years, the prevalence of hypospadias and whether that prevalence is rising or stable has become part of a larger debate over the potential effects of so-called 'endocrine disruptors', such as phthalates and bisphenol-A, on male reproductive health. This commentary critically examines allegations suggesting a worldwide increase in hypospadias rates. Despite the lack of scientific support for this hypothesis and for related claims that the disorders of male reproductive health are related to endocrine disruptors, these constructs remain firmly entrenched in popular literature, and are being used in part to justify litigation banning suspected endocrine disruptors such as phthalates and bisphenol-A. A review of the epidemiologic data on this issue amassed to date clearly demonstrates that the bulk of evidence refutes claims for an increase in hypospadias rates. This suggests that two of the three components of alleged testicular dysfunction syndrome, i.e. decline in sperm counts and rise in urogenital anomalies, lack clinical support. It further suggests that extrapolations from data derived in laboratory animal studies about alleged occult risks to humans of various candidate compounds are unwarranted at this time. PMID- 20022564 TI - Howard Kelly establishes gynecologic brachytherapy in the United States. AB - PURPOSE: Exploration of Howard Atwood Kelly's contributions to gynecologic brachytherapy. METHODS AND MATERIALS: Review of contemporary journals, texts, newspaper accounts, and the memoirs of Kelly's associates. Information from unpublished material, including Kelly's handwritten notes and diaries, was culled from the Alan Mason Chesney Archives of the Johns Hopkins Medical Institutions. RESULTS: Despite European reports of radium's efficacy, gynecologists on both sides of the Atlantic resisted its adoption. The endorsement of radium therapy by America's foremost gynecologist, Howard Kelly, was instrumental in its acceptance. His consummate skill as clinician, investigator, publicist, and entrepreneur established brachytherapy as the primary treatment modality for carcinoma of the cervix and vagina. The technique he pioneered in the second decade of the 20th century, a combination of brachytherapy and megavoltage equivalent teletherapy, presaged modern practice. CONCLUSION: Principles for the management of female genital neoplasia, outlined by Howard Kelly nine decades ago, remain relevant today. PMID- 20022565 TI - The impact of perineural invasion on biochemical outcome after permanent prostate iodine-125 brachytherapy. AB - PURPOSE: Perineural invasion (PNI) in prostate biopsies is associated with increased risk of higher Gleason score and worse pathologic stage. We report the influence of PNI in biochemical no evidence of disease (bNED) survival after (125)I prostate brachytherapy (BT). METHODS AND MATERIALS: Pathology reports of 700 men with localized prostate cancer who underwent (125)I prostate BT in 1999 2008 were reviewed. The presence or absence of PNI in the biopsy was documented in 339 men. Clinical, treatment, and dosimetric parameters, along with PNI status, were evaluated for bNED survival, defined by "nadir+2" definition. RESULTS: Of the 339 patients, 87% had favorable risk and 13% intermediate risk. PNI was present in 89 patients (26%). After a median followup of 32 months, there were five biochemical failures (4: +PNI and 1: -PNI), of which one was local failure (+PNI). Actuarial 5-year bNED survival for the entire group was 97.0% (92.9% for +PNI; 99.2% for -PNI). In univariate analysis age, pretreatment prostate-specific antigen, Gleason score 7, and intermediate risk group predicted for worse biochemical outcome, whereas the presence of PNI showed a trend toward significance (p=0.06). Some of the regression algorithms failed to converge because of low event rates. CONCLUSIONS: We report excellent biochemical control in 339 men treated with (125)I prostate BT. The presence of PNI showed a trend toward significance in predicting 5-year bNED survival but did not impact on local control and should not influence the decision to recommend BT for localized prostate cancer. PMID- 20022566 TI - Simultaneous determination of didanosine and its amino acid prodrug, valdidanosine by hydrophilic interaction chromatography coupled with electrospray ionization tandem mass spectrometry: application to a pharmacokinetic study in rats. AB - A rapid, sensitive and selective ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) method with hydrophilic interaction chromatography has been developed and validated for the simultaneous determination of didanosine and valdidanosine (L-valine amino acid ester prodrug of didanosine) in rat plasma. Solid-phase extraction (SPE) column was employed to extract the analytes from rat plasma, with high extraction recovery (>85%) for both didanosine and valdidanosine. The analytes were then separated by hydrophilic interaction chromatography (HILIC column) and detected by a triple-quadrupole mass spectrometry equipped with an electrospray ionization (ESI) source. The method was linear over the concentration ranges of 2-20,000 ng/mL for didanosine and 4 300 ng/mL for valdidanosine. The lower limit of quantitation (LLOQ) of didanosine and valdidanosine was 2 and 4 ng/mL, respectively. The intra-day and inter-day relative standard deviation (RSD) were less than 15% and the relative errors (RE) were all within 15%. Finally, the validated UPLC-MS/MS method was successfully applied to the pharmacokinetic study after either didanosine or valdidanosine orally administrated to the Sprague-Dawley rats. PMID- 20022567 TI - Methodological issues in the biological monitoring of urinary benzene and S phenylmercapturic acid at low exposure levels. AB - Biological monitoring of low level exposure to pollutants is a very challenging analytical activity, and the quality of results is difficult to assess, especially when a certified reference material is unavailable. The aim of this work was to evaluate the reliability of the assays used to measure urinary benzene (Benz-U) and S-phenylmercapturic acid (SPMA), by applying an internal quality control protocol. Urine spot samples from 705 subjects who were either members of the general urban population, gasoline station attendants, or refinery plant workers were assayed for Benz-U and SPMA, using GC/MS and LC/MS/MS, with quantification limits of 15 ng/L and 0.10 MUg/L. The median Benz-U concentration was 263 ng/L (60-2789 ng/L, 5th-95th percentile), and the median SPMA concentration was 0.19 MUg/L (<0.1-2.5 MUg/L, 5th-95th percentile). Linearity of both assays was good, but a less-than-proportional response was found for SPMA concentrations below 1 MUg/L. Between-run precision and accuracy for Benz-U concentration determination were assessed using quality controls at 120 ng/L and 1000 ng/L and were 10.3% and 4.8%, and 104.8% and 98.9%, respectively; while the precision and accuracy for SPMA concentration determination at 0.3 MUg/L, 2.5 MUg/L, and 20 MUg/L were 40.3%, 6.2%, and 6.2%, and 48.3%, 96.3%, and 98.8%, respectively. Precision, estimated using duplicates of unknown samples, was 13.4% for Benz-U and 26.5% for SPMA analyses. Control charts for the means of the slope of the linear calibration curve of Benz-U showed good stability of the means over a five-year period. For SPMA, a two-laboratory comparison revealed acceptable agreement between ln-transformed data pairs, with a slope of the linear regression of 0.863 (confidence interval 0.774-0.952), null intercept, and a Pearson's r value of 0.844. Reliable results were obtained for Benz-U analyses over the entire concentration range, and for high and medium SPMA levels. However, the determination of SPMA concentrations at levels close to the limit of quantification was less reliable. PMID- 20022568 TI - Immunoaffinity techniques in analysis. PMID- 20022569 TI - The combined application of advanced endoscopic imaging techniques may increase the duodenal villous morphology definition in suspected celiac disease. PMID- 20022570 TI - [Using newborn congenital hypothyroidism screening specimens to detect iodine deficiency in three regions of Spain]. AB - BACKGROUND: Neonatal hyperthyrotropinemia by measurements of thyrotropin (TSH) concentrations has been assessed in three different areas of Spain. The repercussions of a possible iodine deficiency in a congenital hypothyroidism screening program have also been analysed MATERIAL AND METHODS: The study comprised 113,108 newborns, which were divided into two groups according to the time of blood sampling. In 78,646 newborns heel blood samples were obtained after 48h whereas in 34,462 newborns, samples were obtained at birth from the umbilical cord (early samples). Newborns came from three areas of the south of Spain, Seville, Huelva and Cordoba. TSH concentrations were measured by time-resolved fluoroimmunoassay. RESULTS: The percentage of hyperthyrotropinemia was greater in Huelva (5.2%) than Seville (1.0%) (p<0.001), similar to that observed in early samples, which was higher in Huelva (5.3%) than in Seville (1.9%) and Cordoba (1.7%) (p<0.001). In the early samples group, 0.3% and 0.2% of the newborns from Seville and Cordoba respectively, had TSH >20mIU/L and 10 infants should have been recalled for a new sample for each case detected. While in Huelva 17 infants (0.9%) were recalled per case detected. CONCLUSIONS: The heterogeneous distribution of TSH concentrations in newborns from several geographical areas appears to indicate an irregular and deficient iodine intake. Using early samples and a possible iodine deficiency, increase false positive results in a Neonatal Screening Program of congenital hypothyroidism. PMID- 20022572 TI - [Rasburicase versus allopurinol in the treatment of hyperuricaemia in tumour lysis syndrome]. AB - INTRODUCTION: Hyperuricaemia accompanying tumour lysis syndrome (TLS) is a serious complication in neoplasias with rapid proliferation and cellular destruction. The aim of the study was to evaluate the effects of rasburicase versus allopurinol on plasma uric acid, creatinine and phosphorus levels in paediatric patients with TLS. PATIENTS AND METHOD: A comparative study of treatment with rasburicase or allopurinol was performed in 32 paediatric patients with haematological-oncological malignancies and with established TLS or a high risk of developing it, admitted to the Paediatric Intensive Care Unit (PICU). Allopurinol (10mg/kg/day every 8h) was administered to 16 patients between January 1991 and January 2003, and 16 patients received rasburicase (0.2mg/kg/day, once daily), from February 2003 to June 2009. Plasma uric acid, creatinine and phosphorus levels were measured at baseline and 4, 12, 24, 36, 48, 72 and 96h after therapy in both study groups. RESULTS: Baseline uric acid levels were similar in both groups. Four hours after the first dose, patients treated with rasburicase achieved a greater reduction (p<0.0001) of initial plasma uric acid levels compared to allopurinol, as in the other serial determinations. Creatinine levels were higher in the allopurinol group than in the rasburicase one. Plasma phosphorus levels were similar in both groups. Haemodialysis was required in 56% of patients in the allopurinol group, whereas none of rasburicase group needed this. CONCLUSION: This study demonstrated that there is more rapid control and lower plasma uric acid levels in patients at high risk for tumour lysis syndrome who received rasburicase compared to allopurinol, as well as lower levels of creatinine and a lower percentage of haemodialysis. PMID- 20022573 TI - Seminars in fetal and neonatal medicine "continuing controversies in perinatal jaundice". PMID- 20022574 TI - Exploring the genetic architecture of neonatal hyperbilirubinemia. AB - The potential for genetic variation to modulate neonatal hyperbilirubinemia risk is increasingly being recognized. In particular, polymorphisms across three genes involved in bilirubin production and metabolism [glucose-6-phosphate dehydrogenase (G6PD), uridine diphosphate glucuronosyl transferase 1A1 (UGT1A1), and solute carrier organic anion transporter polypeptide 1B1 (SLCO1B1)] may interact with each other and/or environmental contributors to produce significant hyperbilirubinemia. Variant gene co-expression including compound and synergistic heterozygosity enhances hyperbilirubinemia risk, contributing to the etiologic heterogeneity and complex nature of neonatal jaundice. PMID- 20022571 TI - [Study of variability in the management of acute bronchiolitis in Spain in relation to age of patients. National multicenter study (aBREVIADo project)]. AB - INTRODUCTION: There are many studies on the controversial issues involved in the management of acute bronchiolitis (AB). The aim of this multicenter nationwide study in Spain was to find out the variability in the management of AB in various areas of clinical care (primary care, emergency and hospitalization) and to analyze the impact of patient age on diagnostic and therapeutic management. MATERIAL AND METHODS: A cross-sectional observational study (from October 2007 to March 2008) of all cases of BA (McConnochie criteria) treated in a sample of 31 hospitals and 60 health or primary care centers in 12 autonomous regions in Spain. A questionnaire was designed to collect study variables (general information, symptoms, risk factors, diagnostic tests and treatments) and to make a comparison of variables by age groups (newborns, 1-3 months, 3-12 months and >12 months). RESULTS: A total of 5647 cases of AB (51.2% from emergency services, 28.9% from hospitalization, 18.3% from primary care and 1.6% from ICU), whose average age was 0.34 years (95%CI 0.32 to 0.35), with 6.6% under 1 month, 23.5% between 1 and 3 months, 60.6% from 3 to 12 months and 522 over 12 months. There is an important use of diagnostic and therapeutic procedures not recommended in clinical practice guidelines. We also found differences related to age: younger infants showed an increased risk of admission, clinical differences (greater presence of rhinitis, vomiting, refusal of feedings, apnea and septic appearance and less fever and night cough; more tachypnea, retractions, hypoventilation and impaired consciousness, and lower presence of wheezing), increased number of diagnostic tests (oxygen saturation, chest x-ray, RSV test, CBC, etc.) and differences in patterns of treatment (less use of all medications before diagnosis and the maintenance phase; in the acute phase, increased use of epinephrine and parenteral corticosteroids and lower use of bronchodilators and oral corticosteroids, and more frequent use of supplemental oxygen, intravenous fluids, intravenous antibiotics, nasal washings, respiratory aspiration, chest physiotherapy, assisted ventilation and heliox). CONCLUSIONS: There are discrepancies between routine practice and evidence-based management of AB in Spain (both inpatient and outpatient). There is a high percentage of cases in which diagnostic tests and therapeutic interventions (of unproven effectiveness and usefulness in the BA) are performed, where the use is higher in younger infants. PMID- 20022575 TI - The effect of growth hormone on fibroblast proliferation and keratinocyte migration. AB - BACKGROUND: The beneficial effects of growth hormones (GHs) on wound healing have been reported. Although the mechanism of how GH promotes wound healing is unclear, there are reports showing that the principal factor lies in the GH stimulated production of IGF-1 in topical wounds. In this study, a human primary cell model was devised to examine how the topical application of GHs affects fibroblast proliferation and keratinocyte migration, which play fundamental roles in wound healing. METHODS: The fibroblasts were cultured in media with different concentrations of GH. The amount of fibroblast proliferation was assessed using a tetrazolium-based colourimetric assay (MTT assay). The amount of newly formed IGF I mRNA was measured by reverse transcription and polymerase chain reaction (RT PCR). Keratinocyte migration was compared using a migration assay. RESULTS: Fibroblast proliferation was significantly higher in the experimental group than in the control group (the absorbance of 2.5IU L(-1) GH applied group: 0.3954+/ 0.056, control group: 0.2943+/-0.0554, P<0.05), and the promotion of IGF-I formation by fibroblasts was observed. There was more keratinocyte migration in the experimental group than in the control group (the remaining gap in the 2.5IU L(-1) GH applied group after keratinocyte migration: 46.57+/-2.22% of the primary gap, control group: 75.14+/-3.44%, P<0.05). CONCLUSION: GH enhances the local formation of IGF-1, which activates fibroblast proliferation and keratinocyte migration. These results highlight the potential of the topical application of GHs in the treatment of wounds. PMID- 20022576 TI - Ear dressings post pinnaplasty. PMID- 20022577 TI - The importance of fear, beliefs, catastrophizing and kinesiophobia in chronic low back pain rehabilitation. AB - AIM: To evaluate fear, beliefs, catastrophizing and kinesiophobia in chronic low back pain patients about to begin a training programme in a rehabilitation centre. PATIENTS AND METHODS: Fifty chronic low back pain patients (including both males and females) were assessed in our physical medicine department. We used validated French-language scales to score the patients' pain-related disability, quality of life and psychosocial factors. RESULTS: Seventy percent of the patients had a major functional disability (i.e., a Roland-Morris Scale score over 12) and nearly 73% reported an altered quality of life (the daily living score in the Dallas Pain Questionnaire). Pain correlated with functional impairment and depression but not with catastrophizing or kinesiophobia. Disability was correlated with catastrophizing and kinesiophobia. CONCLUSION: Psychosocial factors are strongly associated with disability and altered quality of life in chronic low back pain patients. Future rehabilitation programs could optimizing patient management by taking these factors into account. PMID- 20022578 TI - The psychological aspects of polio survivors through their life experience. AB - OBJECTIVES: After several decades of stability, polio survivors often experience new signs and symptoms of their condition, characterized by global and muscular fatigue, decreased muscular strength and pain. The hypothesis of a potential underlying psychological component has been suggested. METHOD: This article aims to report how polio survivors cope with their condition based on an analysis of the literature and the authors' experience. RESULTS: The literature does not report a higher prevalence of psychological disorders (mood disorders, depression) in polio survivors than in the general population. Conversely, the psychological impact of the patients' decreasing abilities, regardless of the cause, is regularly reported. Most patients report a restricted involvement in their daily life activities. DISCUSSION AND CONCLUSION: It is essential to be aware of the patients' experience in coping with polio as children as well as the medical treatments they went through in order to better understand their various types of complaints and sometimes their reluctance to go back to a medical environment. With such widespread information on post-polio syndrome (PPS) and all the uncertainties surrounding it, patients fear a late onset deterioration of their condition. In order to provide the best therapeutic advice, it is essential to have a good knowledge of the symptoms but also take the time and listen to patients in order to differentiate the underlying physical and psychological components in symptoms of pain and fatigue. PMID- 20022579 TI - The effect of a chlorhexidine-based surgical lubricant during pelvic examination on the detection of group B Streptococcus. AB - OBJECTIVE: The objective of the study was to estimate whether surgical lubricant used during pelvic examination alters the detection of group B Streptococcus (GBS). STUDY DESIGN: We conducted a prospective cohort study of patients undergoing GBS screening at the prenatal clinics of a New York City public hospital. Two specimens were collected from each patient, before and after a pelvic examination with Surgilube (Fougera and Co, Melville, NY), a bacteriostatic surgical lubricant. Test performance indices using GBS status pre pelvic examination as the reference were calculated. RESULTS: Over 10 months, 168 patients were enrolled in the study. Twenty of 168 patients (11.9%; 95% confidence interval, 7.4-17.8%) tested GBS positive before the pelvic examination. Of the initial 20 GBS-positive patients, 10 tested GBS positive after the pelvic examination with surgical lubricant. The sensitivity of detecting GBS after the examination with surgical lubricant was 50%. CONCLUSION: Because pelvic examination with surgical lubricant may decrease the detection of GBS, obstetric practitioners should collect GBS screening cultures before the use of surgical lubricant. PMID- 20022581 TI - Endometrial sampling diagnosis of FIGO grade 1 endometrial adenocarcinoma with a background of complex atypical hyperplasia and final hysterectomy pathology. AB - OBJECTIVE: The purpose of this study was to determine posthysterectomy pathologic findings in patients with a preoperative endometrial sampling diagnosis of International Federation of Gynecology and Obstetrics (FIGO) grade 1 endometrial adenocarcinoma with a background of complex atypical hyperplasia (CAH). STUDY DESIGN: We reviewed 1423 consecutive cases of endometrial cancer to identify cases with a preoperative endometrial biopsy that demonstrated FIGO grade 1 endometrial adenocarcinoma. Final uterine pathologic findings were grouped into low- and high-risk based on FIGO and Gynecologic Oncology Group criteria. RESULTS: We identified 123 cases with a background of CAH and 367 cases without a background of CAH. FIGO grade in the hysterectomy specimen was more than FIGO grade 1 in 11 of 123 cases (8.9%) with a background of CAH, compared with 60 of 359 cases (16.7%) without a background of CAH (P = .04). CONCLUSION: An endometrial sampling diagnosis of FIGO grade 1 endometrial adenocarcinoma with a background of CAH is more likely to correlate with final posthysterectomy grade than a diagnosis not arising with a background of CAH. PMID- 20022582 TI - Anterior abdominal wall nerve and vessel anatomy: clinical implications for gynecologic surgery. AB - OBJECTIVE: We sought to describe relationships of clinically relevant nerves and vessels of the anterior abdominal wall. STUDY DESIGN: The ilioinguinal and iliohypogastric nerves and inferior epigastric vessels were dissected in 11 unembalmed female cadavers. Distances from surface landmarks and common incision sites were recorded. Additional surface measurements were taken in 7 other specimens with and without insufflation. RESULTS: The ilioinguinal nerve emerged through the internal oblique: mean (range), 2.5 (1.1-5.1) cm medial and 2.4 (0 5.3) cm inferior to the anterior superior iliac spine (ASIS). The iliohypogastric emerged 2.5 (0-4.6) cm medial and 2.0 (0-4.6) cm inferior. Inferior epigastric vessels were 3.7 (2.6-5.5) cm from midline at the level of the ASIS and always lateral to the rectus muscles at a level 2 cm superior to the pubic symphysis. CONCLUSION: Risk of anterior abdominal wall nerve and vessel injury is minimized when lateral trocars are placed superior to the ASISs and >6 cm from midline and low transverse fascial incisions are not extended beyond the lateral borders of the rectus muscles. PMID- 20022583 TI - Effects of recommended levels of physical activity on pregnancy outcomes. AB - OBJECTIVE: We sought to examine the relation between recommended levels of physical activity during pregnancy and pregnancy outcomes. STUDY DESIGN: We conducted an observational study with energy expenditure, aerobic fitness, and sleeping heart rate measured in 44 healthy women in late pregnancy. Medical records were examined for pregnancy outcome. RESULTS: Active women, who engaged in > or = 30 minutes of moderate physical activity per day, had significantly better fitness and lower sleeping heart rate compared to the inactive. Duration of second stage of labor was 88 and 146 minutes in the active vs inactive women, respectively (P = .05). Crude odds ratio of operative delivery in the inactive vs the active was 3.7 (95% confidence interval, 0.87-16.08). Birthweight, maternal weight gain, and parity adjusted odds ratio was 7.6 (95% confidence interval, 1.23-45.8). Neonatal condition and other obstetric outcomes were similar between groups. CONCLUSION: Active women have better aerobic fitness as compared to inactive women. The risk for operative delivery is lower in active women compared to inactive, when controlled for birthweight, maternal weight gain, and parity. Further studies with larger sample size are required to confirm the association between physical activity and pregnancy outcomes. PMID- 20022585 TI - Extraabdominal vs intraabdominal uterine repair at cesarean delivery: a metaanalysis. PMID- 20022586 TI - Clinical differences between early-onset HELLP syndrome and early-onset preeclampsia during pregnancy and at least 6 months postpartum. AB - OBJECTIVE: We sought to evaluate whether clinical and laboratory variables differ between former patients who had HELLP syndrome and former patients who had preeclampsia (PE) without HELLP. STUDY DESIGN: We compared early-onset HELLP (n = 75) with early-onset PE (n = 40) with respect to clinical features during the hypertensive complication and to metabolic, hemodynamic, and hemostatic variables determined at least 6 months postpartum. RESULTS: HELLP differed from PE by a borderline higher frequency of eclampsia (13% vs 3%) during the complication, and by a lower prevalence of hypertension (19% vs 33%), proteinuria (2% vs 23%), thrombophilia (6% vs 27%), obesity (9% vs 33%), hypertriglyceridemia (1% vs 15%), hyperglycemia (0% vs 11%), and elevated levels of fasting homocysteine (6% vs 21%) at least 6 months postpartum. CONCLUSION: Women with HELLP had fewer signs of abnormalities consistent with the metabolic syndrome and a 4-fold lower prevalence of thrombophilia as compared with PE women without HELLP. PMID- 20022587 TI - Early menstrual characteristics associated with subsequent diagnosis of endometriosis. AB - OBJECTIVE: The aim of this study was to investigate the association between early menstrual characteristics, before symptom onset, and later diagnosis of endometriosis. STUDY DESIGN: This was a case-control study of 268 Australian women with surgically confirmed moderate-to-severe endometriosis (cases) and 244 women without endometriosis (controls). Early menstrual cycle characteristics, before age at symptom onset, were analyzed. RESULTS: Menarche after age 14 years was strongly and inversely associated with endometriosis (odds ratio, 0.3; 95% confidence interval, 0.1-0.6). A history of dysmenorrhea was associated with subsequent endometriosis (odds ratio, 2.6; 95% confidence interval, 1.1-6.2). Despite a suggestive trend, shorter menstrual cycle length was not associated with endometriosis. Duration of natural menstruation and heaviness of flow were not associated with subsequent risk of endometriosis; neither was the reported type of sanitary protection used nor history of sexual intercourse during menstruation. CONCLUSION: There is a decreased risk of endometriosis with late age at menarche and an increased risk in women who report an early history of dysmenorrhea. PMID- 20022588 TI - Pelvic pain and surgeries in women before interstitial cystitis/painful bladder syndrome. AB - OBJECTIVE: The objective of the study was to compare subjects with interstitial cystitis/painful bladder syndrome (IC/PBS) with controls on prior surgeries. STUDY DESIGN: IC/PBS subjects were compared with matched controls on surgeries and possible surgical indications prior to their index dates. RESULTS: Adjusted for demographic variables, logistic regression showed subjects exceeded controls in surgeries longer than 12 months and less than 1 month before the index date. However, addition of possible surgical indications showed chronic pelvic pain (CPP) to have a strong association with IC/PBS, whereas associations with surgeries were reduced to nonsignificance. CONCLUSION: Although women with IC/PBS were more likely to have experienced prior surgeries than controls, the apparent indications for surgeries, not the surgeries themselves, were stronger risk factors for IC/PBS. In particular, a prior history of CPP had a strong association with IC/PBS. Several features of study design, including extensive medical record review, suggest that prior CPP was not undiagnosed IC/PBS. Further investigation of CPP may yield insight into the pathogenesis of IC/PBS. PMID- 20022590 TI - Mom told me scary things about this animal: parents installing fear beliefs in their children via the verbal information pathway. AB - This study investigated whether fear beliefs can be installed in children after parents had received negatively tinted information about a novel stimulus. Parents of children aged 8-13 years (N = 88) were presented with negative, positive, or ambiguous information about an unknown animal and then given a number of open-ended vignettes describing confrontations with the animal with the instruction to tell their children what would happen in these situations. Results indicated that children's fear beliefs were influenced by the information that was provided to the parent. That is, parents who had received negative information provided more threatening narratives about the animal and hence installed higher levels of fear beliefs in their children than parents who had received positive information. In the case of ambiguous information, the transmission of fear was dependent on parents' trait anxiety levels. More precisely, high trait anxious parents told more negative stories about the unknown animal, which produced higher fear levels in children. PMID- 20022591 TI - Two-dimensional genome scan identifies multiple genetic interactions in bipolar affective disorder. AB - BACKGROUND: Bipolar disorder is a highly heritable psychiatric condition, the etiology of which remains largely unknown despite extensive efforts to identify susceptibility genes. Interactions between genes of small individual effect could partially explain the difficulties of traditional one-dimensional approaches to identify genetic risk factors. METHODS: A nonparametric linkage (NPL) analysis of 65 Australian extended pedigrees containing 643 genotyped individuals (of whom 40% were diagnosed with affective disorder) was conducted. Chromosome-by chromosome correlation analysis of family-specific NPL scores was conducted to detect evidence of genetic interaction. Interaction-specific multipoint NPL and permutation analysis was used to assess linkage interdependence, using family weights derived from the alternative interacting chromosome. Finally, a single nucleotide analysis of each interaction region was conducted using the publicly available genome-wide association, datasets (2933 cases, 2534 controls). RESULTS: Significant NPL peaks were detected on chromosomes 2q24-33, 7q21-31, and 17q11-25 (Z = 3.12, 3.01, and 2.95 respectively), with four additional suggestive peaks identified. Four robust interchromosomal interaction clusters exceeding Bonferroni correction at alpha = .05 (uncorrected p < 5.38e-07) were detected on 11q23-25-2p15-12, 4q32-35-1p36, 12q23-24-4p16-15, and 20q13-9q21-22. This linkage interdependence was determined significant after permutation analysis (p = .002 .0002). A suggestive interaction was observed in the combined data on 2p14-11q23 (uncorrected p = 5.76E-10, Bonferroni corrected p = .068). CONCLUSIONS: This study indicates a complex interplay between multiple loci underlying bipolar disorder susceptibility, and highlights the continuing usefulness of extended pedigrees in complex genetics. The challenge lies in the identification of specific gene interactions and their biological validation. PMID- 20022593 TI - Heterologous CD8 T cell immune response to HSV induced by toll like receptor ligands. AB - A memory response is established following primary antigen exposure that stays more or less constant. It appears to adopt a set-point in magnitude but upon re exposure the response is quicker and better and there is an upward shift in memory frequency that varies with individuals based on the exposure pattern to other microbes or its components. Our investigations were designed to test such differences of non-specific stimulation by PAMPs in lowering the threshold of activation. Neonatal mice were pre-exposed to TLR-ligands intermittently and later analyzed for its resilience to challenge with virus during adult-life. Secondly, adult mice with pre-existing memory to virus were exposed to various TLR-ligands and analyzed for their quality of memory response. The TLR-ligands exposed animals were better responders to a new agent exposure compared to the animals kept in sterile surroundings. Moreover, immune memory recall and the viral specific CD8(+) T cells response with TLR-ligands were comparable to the recall response with the cognate antigen. The results provide insights into the role of hyper-sanitized environment versus PAMPs mediated signaling in adaptive immunity and long-term immune memory. PMID- 20022594 TI - Grasping spheres, not planets. AB - Memory for objects helps us to determine how we can most effectively and appropriately interact with them. This suggests a tightly coupled interplay between action and background knowledge. Three experiments demonstrate that grasping circumference can be affected by the size of a visual stimulus (Experiment 1), whether that stimulus appears to be graspable (Experiment 2), and the presence of a label that renders that object ungraspable (Experiment 3). The results are taken to inform theories on conceptual representation and the functional distinction that has been drawn between the visual systems for perception and action. PMID- 20022592 TI - A novel role for glyceraldehyde-3-phosphate dehydrogenase and monoamine oxidase B cascade in ethanol-induced cellular damage. AB - BACKGROUND: Alcoholism is a major psychiatric condition at least partly associated with ethanol (EtOH)-induced cell damage. Although brain cell loss has been reported in subjects with alcoholism, the molecular mechanism is unclear. Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and monoamine oxidase B (MAO B) reportedly play a role in cellular dysfunction under stressful conditions and might contribute to EtOH-induced cell damage. METHODS: Expression of GAPDH and MAO B protein was studied in human glioblastoma and neuroblastoma cell lines exposed to physiological concentrations of EtOH. Expression of these proteins was also examined in the prefrontal cortex from human subjects with alcohol dependence and in rats fed with an EtOH diet. Coimmunoprecipitation, subcellular fractionation, and luciferase assay were used to address nuclear GAPDH-mediated MAO B activation. To test the effects of inactivation, RNA interference and pharmacological intervention were used, and cell damage was assessed by terminal deoxynucleotidyl transferase (TdT)-mediated dUTP Nick End Labeling (TUNEL) and hydrogen peroxide measurements. RESULTS: Ethanol significantly increases levels of GAPDH, especially nuclear GAPDH, and MAO B in neuronal cells as well as in human and rat brains. Nuclear GAPDH interacts with the transcriptional activator, transforming growth factor-beta-inducible early gene 2 (TIEG2), and augments TIEG2-mediated MAO B transactivation, which results in cell damage in neuronal cells exposed to EtOH. Knockdown expression of GAPDH or treatment with MAO B inhibitors selegiline (deprenyl) and rasagiline (Azilect) can block this cascade. CONCLUSIONS: Ethanol-elicited nuclear GAPDH augments TIEG2-mediated MAO B, which might play a role in brain damage in subjects with alcoholism. Compounds that block this cascade are potential candidates for therapeutic strategies. PMID- 20022595 TI - Vascular intersection detection in retina fundus images using a new hybrid approach. AB - The use of vascular intersection aberration as one of the signs when monitoring and diagnosing diabetic retinopathy from retina fundus images (FIs) has been widely reported in the literature. In this paper, a new hybrid approach called the combined cross-point number (CCN) method able to detect the vascular bifurcation and intersection points in FIs is proposed. The CCN method makes use of two vascular intersection detection techniques, namely the modified cross point number (MCN) method and the simple cross-point number (SCN) method. Our proposed approach was tested on images obtained from two different and publicly available fundus image databases. The results show a very high precision, accuracy, sensitivity and low false rate in detecting both bifurcation and crossover points compared with both the MCN and the SCN methods. PMID- 20022589 TI - The effect of maternal body mass index on neonatal outcome in women receiving a single course of antenatal corticosteroids. AB - OBJECTIVE: The aim of this study was to determine the effect of maternal body mass index on the incidence of neonatal prematurity morbidities in those who receive corticosteroids. STUDY DESIGN: This was a secondary analysis of a trial of corticosteroids in women at risk for preterm birth. Women receiving a single course of corticosteroids were classified by their prepregnancy body mass index (<25 and > or = 25) and compared on a composite outcome comprised of several neonatal morbidities and on each individual outcome. RESULTS: Of 183 eligible women, 96 (52.5%) had a body mass index of <25 and 87 (47.5%) had a body mass index of > or = 25. The composite outcome occurred more frequently in the body mass index of > or = 2 5 group (28.7%), compared with those with a body mass index of <25 (18.8%), although this was not statistically significant (odds ratio, 1.75; 95% confidence interval, 0.83-3.72). Body mass index was not associated with outcomes after adjusting for confounding. CONCLUSION: Maternal body mass index did not affect neonatal prematurity morbidities in those receiving corticosteroids. PMID- 20022596 TI - Multiclass detection of cells in multicontrast composite images. AB - In this paper, we describe a framework for multiclass cell detection in composite images consisting of images obtained with three different contrast methods for transmitted light illumination (referred to as multicontrast composite images). Compared to previous multiclass cell detection results [1], the use of multicontrast composite images was found to improve the detection accuracy by introducing more discriminatory information into the system. Preprocessing multicontrast composite images with Kernel PCA was found to be superior to traditional linear PCA preprocessing, especially in difficult classification scenarios where high-order nonlinear correlations are expected to be important. Systematic study of our approach under different overlap conditions suggests that it possesses sufficient speed and accuracy for use in some practical systems. PMID- 20022597 TI - Pelvic hydatid (echinococcal) disease. AB - OBJECTIVE: To study epidemiologic and clinical features of pelvic hydatid disease and discuss its management. METHOD: A retrospective analysis of 11 cases of pelvic hydatid disease managed over 7 years and 8 months at the Maternity and Neonatalogy Unit, Tunisian Medical Center La Rabta, Tunis, Tunisia. All cases were identified from histopathologic reports. RESULTS: The 11 affected patients had a mean age of 41.6years (range, 22-79 years), 6 had a history of surgery for hydatid disease, 8 presented for chronic pelvic pain, and 1 was admitted for acute surgical abdomen. On physical examination, 6 had a pelvic mass. An ultrasound examination suggested the diagnosis preoperatively in 6. All were treated surgically. Primary laparoscopy was performed in 5 patients. Unroofing (or partial cystectomy) was performed in 6 patients and complete cystectomy in 4. The postoperative course was uneventful in all cases. Recurrence occurred only in 1 patient, 6 months after initial surgery. CONCLUSION: Pelvic hydatid disease is rare and its diagnosis often difficult preoperatively. The treatment mainstay is surgery. The laparoscopic approach seems to be safe and effective, and may increasingly replace laparotomy. PMID- 20022599 TI - Consequences of delay in obstetric care for maternal and perinatal outcomes. PMID- 20022598 TI - "Top hat" versus conventional loop electrosurgical excision procedure in women with a type 3 transformation zone. AB - OBJECTIVE: To compare the "top-hat" and conventional loop electrosurgical excision procedures (LEEP) performed in women with a type 3 transformation zone to assess the rate of endocervical margin involvement. METHODS: Women with a type 3 transformation zone randomly allocated into the conventional (n=94) and top-hat LEEP (n=86) groups were analyzed. RESULTS: The rate of endocervical margin involvement in the top-hat group was lower than that in the conventional group (32.6% vs 53.2%; RR 0.36; 95% CI, 0.19-0.68; P=0.003). Among women with positive endocervical margins, women undergoing top-hat LEEP were less likely to have residual lesions compared with those in the conventional group (52.2% vs 84.1%, respectively, P=0.04). There was no significant difference in the complication rate between the top-hat and conventional groups (7.0% vs 10.6%, respectively, P=0.39). CONCLUSION: Top-hat LEEP performed in women with a type 3 transformation zone reduces the risks of endocervical margin involvement and residual diseases compared with conventional LEEP, with no significant difference in perioperative complications. PMID- 20022600 TI - A simple procedure to prevent chronic vaginal colpotomy wound bleeding after laparoscopically assisted vaginal hysterectomy. AB - OBJECTIVE: To determine whether injecting the colpotomy wound with diluted vasopressin decreases vaginal bleeding after laparoscopically assisted vaginal hysterectomy (LAVH). METHODS: In this prospective controlled study 100 patients who underwent LAVH from July 1, 2005 to June 30, 2007, were randomized to receive an injection of vasopressin (n=50) or normal saline (n=50) solution in the colpotomy wound. RESULTS: In the vasopressin group, bleeding from the colpotomy wound occurred for more than 7 days in 9 patients (18%), and none was bleeding after 1 month; in the control group, the corresponding values were 29 (58%) and 2 (4%). Compared with the study group, the control group had a significantly higher rate of chronic bleeding from the colpotomy wound for more than 7 days and for more than 14 days after LAVH (P<0.001 for both). CONCLUSION: Infiltrating the colpotomy wound with diluted vasopressin was found to prevent chronic vaginal bleeding, which frequently occurs following LAVH. PMID- 20022601 TI - Development and validation of an instrument to measure nurse-patient bonding. AB - BACKGROUND: Most instruments on nurse-patient relationship determine the caring behavior of the nurse, but have minimal consideration of the patient's role in the interaction. Moreover, it is the patients that complete many of those instruments, thus leaving out the perspective of the nurse. There is then a need to account for the contributions that both the nurse and the patient bring into their encounter where bonding is formed. OBJECTIVE: This study aimed to develop and validate an instrument that determines the degree of bonding between nurse and patient based on their openness to each other and their engagement in patient care. SETTINGS: Data were collected from nurses and patients in the wards of four public and private tertiary hospitals in Manila, Philippines, where most Filipino nurses render care to patients before getting employed in other countries. PARTICIPANTS: A total of 420 nurses and patients (i.e., 210 dyads) participated in this research conducted in 2008. Most of the nurses were young females with beginning clinical experience, while the patients had a wider age range with the majority having no college education and no employment. METHODS: The Nurse Patient Bonding Instrument (NPBI), which dimensions were generated from qualitative observations and interviews, and corroborated by literature, was validated at the bedside setting. To determine interrater reliability, two trained raters unobtrusively observed actual nurse-patient interactions and ticked on the NPBI behavioral indicators of openness and engagement. Construct validity was established using known-groups technique. Moreover, bonding score was correlated with patient satisfaction for predictive validity. RESULTS: Reliability ranged from r=.80 to .95 (p<.01). Factor analysis demonstrated that the subscale scores of patient openness, nurse openness, patient engagement, and nurse engagement all loaded on one factor, the bonding factor, demonstrating a unified structure of the NPBI. Nurses and patients had higher bonding scores in interactions of longer duration than shorter duration, controlling for number of previous encounters. This provided evidence for construct validity using known groups technique. The NPBI was likewise shown to distinguish groups based on age, education, and civil status. Patient satisfaction correlated positively with bonding score, providing evidence suggestive of the predictive validity of the NPBI. CONCLUSION: The NPBI was shown to be a reliable and valid tool for assessing nurse-patient bonding, and can possibly predict patient satisfaction. The openness and engagement of nurse and patient were demonstrated to result in a structure, a nurse-patient dyad. This finding invites further investigations on the characteristics and development of this dyad. PMID- 20022602 TI - Endothelium oriented differentiation of bone marrow mesenchymal stem cells under chemical and mechanical stimulations. AB - Bone marrow mesenchymal stem cells (MSCs) have multi-differentiation capability. Their endothelial cell (EC) oriented differentiation is the key to vasculogenesis, in which both mechanical and chemical stimulations play important roles. Most previous studies reported individual effects of VEGF or fluid shear stress (SS), when MSCs were subjected to shear stress of 10-15 dyn/cm(2) over 24hr. In this paper, we investigated responses of MSCs from young Sprague Dawley rats to shear stress, VEGF and the combination of the two stimuli. Our study showed that the combined stimulation of shear stress and VEGF resulted in more profound EC oriented differentiation of MSCs in comparison to any individual stimulation. Furthermore, we subjected MSCs to prolonged period of fluid shear stimulation, i.e. 48 hr rather than 24hr, and increased the magnitude of the shear stress from 10 dyn/cm(2) to 15, 20 and 25 dyn/cm(2). We found that without VEGF, the endothelium oriented differentiation of MSCs that was seen following 24hr of shear stimulation was largely abolished if we extended the shear stimulation to 48hr. A similar sharp decrease in MSC differentiation was also observed when the magnitude of the shear stress was increased from 10-15 dyn/cm(2) to 20-25 dyn/cm(2) in 24hr shear stimulation studies. However, with combined VEGF and fluid shear stimulation, most of the endothelial differentiation was retained following an extended period, i.e. at 48 hr, of shear stimulation. Our study demonstrates that chemical and mechanical stimulations work together in determining MSC differentiation dynamics. PMID- 20022603 TI - Pretreatment-free fast ultraviolet detection of melamine in milk products with a disposable microfluidic device. AB - A new method for sensitive and fast screening of melamine (MEL) in milk products was developed with a low-cost disposable microfluidic device coupled with ultraviolet (UV) detection. This method avoided the need of sample pretreatment prior to the separation process, thus was simple and green. Due to the advantages of the device and fracture sampling technique, milk sample could be directly sampled through the fracture to achieve baseline separation from amino acids, and proteins in the sample did not interfere with the detection. Using 20mM phosphate running buffer (pH 9.0), a sampling time of 3s at +180V and a separation voltage of +1800V (240V/cm), this method could detect MEL in milk within 75s. At the detection wavelength of 202nm, the linear range for MEL was from 1.0 to 100microgmL(-1) with a detection limit of 0.23microgmL(-1) (S/N=3). The novel protocol had been successfully used to screen MEL in milk samples with recovery more than 82%. The environmentally friendly methodology for pretreatment-free sensitive screening of MEL provided promising applications in monitoring the quality of different foods. PMID- 20022604 TI - 6th Hydrophobic interaction chromatography/reversed-phase liquid chromatography bioseparation conference. Foreword. PMID- 20022605 TI - Compact adaptive-grid scheme for high numerical resolution simulations of isotachophoresis. AB - In a previous publication we demonstrated a fast simulation tool for solution of electrophoretic focusing and separation. We here describe the novel mathematical model and numerical algorithms used to create this code. These include the representation of advection-diffusion equations on an adaptive grid, high resolution discretization of the equations (sixth order compact), a new variational-based approach for controlling the motion of grid points, and new boundary conditions which enable solution in a moving frame of reference. We discuss the advantages of combining a high-resolution discretization with an adaptive grid in accurately resolving sharp interfaces in isotachophoresis, and provide verification against known analytical solutions and comparison with prevailing exiting numerical algorithms. PMID- 20022606 TI - Improved extraction and identification by ultra performance liquid chromatography tandem mass spectrometry of phenolic compounds in burdock leaves. AB - The simultaneous ultrasonic and microwave assisted extraction (UMAE) technique was first employed to obtain phenolics. The effects of UMAE variables including extraction time, microwave power, and solvent to solid radio on the yield of phenolics were investigated. The optimized conditions were as follows: solvent to solid ratio was 20:1 (ml/g), extraction time was 30s, microwave power was 500W and two times of extraction. Moreover, the phenolic yield of UMAE was higher than that by maceration, indicating a significant reduction of extraction time and an improvement of efficiency. The phenomenon is related to the strong disruption of leaf tissue structure by microwave induced expansion and ultrasonic shaking, which had been observed with the scanning electron microscopy. The phenolic compositions of the extract was then identified by ultra performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS), 10 compounds had been characterized, providing a more complete identification of phenolic compounds in burdock leaves than previously reported. The occurrence of benzoic acid and p coumaric acid is reported for the first time. This study suggests that UMAE is a good alternative for the extraction of phenolics, with a great potential for industrial application. Also, UMAE provides a new sample preparation technique for characterization of the phenolic compounds from plants. PMID- 20022607 TI - Slowly progressive encephalopathy with hearing loss due to a mutation in the mtDNA tRNA(Leu(CUN)) gene. AB - Pathogenic mutations in the tRNA(Leu(UCN)) gene of mitochondrial DNA (mtDNA) have been invariably accompanied by skeletal myopathy with or without chronic progressive external ophthalmoplegia (CPEO). We report a young woman with a heteroplasmic m.12276G>A mutation in tRNA(Leu(UCN)), who had childhood-onset and slowly progressive encephalopathy with ataxia, cognitive impairment, and hearing loss. Sequencing of the 22 tRNA mitochondrial genes is indicated in all unusual neurological syndromes, even in the absence of maternal inheritance. PMID- 20022608 TI - Association of hospital coronary artery bypass volume with processes of care, mortality, morbidity, and the Society of Thoracic Surgeons composite quality score. AB - OBJECTIVE: This study examines the association of hospital coronary artery bypass procedural volume with mortality, morbidity, evidence-based care processes, and Society of Thoracic Surgeons composite score. METHODS: The study population consisted of 144,526 patients from 733 hospitals that submitted data to the Society of Thoracic Surgeons Adult Cardiac Database in 2007. End points included use of National Quality Forum-endorsed process measures (internal thoracic artery graft; preoperative beta-blockade; and discharge beta-blockade, antiplatelet agents, and lipid drugs), operative mortality (in-hospital or 30-day), major morbidity (stroke, renal failure, reoperation, sternal infection, and prolonged ventilation), and Society of Thoracic Surgeons composite score. Procedural volume was analyzed as a continuous variable and by volume strata (< 100, 100-149, 150 199, 200-299, 300-449, and > or = 450). Analyses were performed with logistic and multivariate hierarchical regression modeling. RESULTS: Unadjusted mortality decreased across volume categories from 2.6% (< 100 cases) to 1.7% (> 450 cases, P < .0001), and these differences persisted after risk factor adjustment (odds ratio for lowest- vs highest-volume group, 1.49). Care processes and morbidity end points were not associated with hospital procedural volume except for a trend (P = .0237) toward greater internal thoracic artery use in high-volume hospitals. The average composite score for the lowest volume (< 100 cases) group was significantly lower than that of the 2 highest-volume groups, but only 1% of composite score variation was explained by volume. CONCLUSION: A volume performance association exists for coronary artery bypass grafting but is weaker than that of other major complex procedures. There is considerable outcomes variability not explained by hospital volume, and low volume does not preclude excellent performance. Except for internal thoracic artery use, care processes and morbidity rates were not associated with volume. PMID- 20022610 TI - Re: Testicular dysfunction with maldescent--can urologists change this with earlier orchiopexy? T. T. Turner J Urol 2009; 182: 429-430. PMID- 20022611 TI - Re: Modern technique for penile torsion repair R. Bauer and B. A. Kogan J Urol 2009; 182: 286-291. PMID- 20022612 TI - Re: High power diode laser vaporization of the prostate: preliminary results for benign prostatic hyperplasia A. Erol, K. Cam, A. Tekin, O. Memik, S. Coban And Y. Ozer J Urol 2009; 182: 1078-1082. PMID- 20022613 TI - [Effectiveness of lumbar sympathectomy in the treatment of occlusive peripheral vascular disease in lower limbs: systematic review]. AB - BACKGROUND AND OBJECTIVES: The aim of this work has been to check the scientific evidence on the efficacy of lumbar sympathectomy in the treatment of peripheral arterial disease of lower limbs. MATERIAL AND METHODS: Systematic review of the literature. DATA SOURCE: MedLine, Embase, Cochrane Library, INHATA, CRD, BioMed Central, and ClinicalTrials.gov register. Mesh terms: sympathectomy, peripheral vascular disease, lower extremity, vascular therapy, iliac artery, femoral artery, popliteal artery. INCLUSION CRITERIA: adult patients with occlusive peripheral arterial disease of lower limbs treated with sympathectomy. The quality was evaluated by means of the criteria of the CASP checklist and "ad hoc" questionnaire. RESULTS: Four clinical trials and four observational studies were included. Sympathectomy did not show significant differences for mortality, amputations and grade of intermittent claudication. When it was assessed regarding local anaesthetics or prostaglandin-E1, differences were not detected. Chemical sympathectomy showed better results than surgical sympathectomy in hospital stance. CONCLUSIONS: Evidence on the efficacy of sympathectomy does not show differences related to conventional treatments. Both sympathectomy techniques (surgical and chemical) do not report statistical differences regarding mortality or amputations. PMID- 20022614 TI - Anti-annexin A5 antibodies in women with spontaneous pregnancy loss. AB - BACKGROUND AND OBJECTIVE: The aim was to evaluate the role of anti-annexin A5 (anti-ANXA5) antibodies as risk factor for recurrent miscarriage (RM) and unexplained fetal loss (UFL). PATIENTS AND METHODS: Retrospective, cohort study. SETTING: Vall d'Hebron University Hospital. SUBJECTS: 122 women, in two groups: STUDY GROUP: 54 women with RM/UFL and control group: 68 pregnant without RM/UFL. INTERVENTION: Antiphospholipid, mainly anti-ANXA5 antibody analysis. Comparison of all antiphospholipid antibodies between groups. RESULTS: Antiphospholipid antibody (aPL) prevalence in the study group was 10/54 (14.8%) and 5/68 (7.3%) in control group (p=0.09). In the RM subgroup, it was 3/25 and 9/34 in UFL versus 5/68 in controls (p=0.013). Lupus anticoagulant (LA) was present in 4 cases, all belonging to the study group (p=0.011). Four out of 34 women with UFL were positive for anticardiolipin antibodies-IgG (IgG-aCL) versus 1/68 in controls (p=0.041). In RM subgroup, anti-ANXA5 antibodies were positive in 2/25 versus 3/68 in controls, and in UFL subgroup, 3/34 versus 3/68 cases (p=1.000). CONCLUSION: According to our results, anti-ANXA5 antibodies should not be considered as a risk factor for RM/UFL. PMID- 20022615 TI - [The research projects in health of the Spanish Sanitary Research Fund in the year 2008: accordance to the VI National Plan of R&D 2008-2011]. PMID- 20022616 TI - Characterization of the metabolic and physiologic response to chromium supplementation in subjects with type 2 diabetes mellitus. AB - The objective of the study was to provide a comprehensive evaluation of chromium (Cr) supplementation on metabolic parameters in a cohort of type 2 diabetes mellitus subjects representing a wide phenotype range and to evaluate changes in "responders" and "nonresponders." After preintervention testing to assess glycemia, insulin sensitivity (assessed by euglycemic clamps), Cr status, and body composition, subjects were randomized in a double-blind fashion to placebo or 1000 microg Cr. A substudy was performed to evaluate 24-hour energy balance/substrate oxidation and myocellular/intrahepatic lipid content. There was not a consistent effect of Cr supplementation to improve insulin action across all phenotypes. Insulin sensitivity was negatively correlated to soleus and tibialis muscle intramyocellular lipids and intrahepatic lipid content. Myocellular lipids were significantly lower in subjects randomized to Cr. At preintervention, responders, defined as insulin sensitivity change from baseline of at least 10% or greater, had significantly lower insulin sensitivity and higher fasting glucose and A(1c) when compared with placebo and nonresponders, that is, insulin sensitivity change from baseline of less than 10%. Clinical response was significantly correlated (P < .001) to the baseline insulin sensitivity, fasting glucose, and A(1c). There was no difference in Cr status between responder and nonresponders. Clinical response to Cr is more likely in insulin-resistant subjects who have more elevated fasting glucose and A(1c) levels. Chromium may reduce myocellular lipids and enhance insulin sensitivity in subjects with type 2 diabetes mellitus who do respond clinically independent of effects on weight or hepatic glucose production. Thus, modulation of lipid metabolism by Cr in peripheral tissues may represent a novel mechanism of action. PMID- 20022617 TI - Dihydrochalcones: Implication in resistance to oxidative stress and bioactivities against advanced glycation end-products and vasoconstriction. AB - Flavonoids are a group of polyphenol compounds with known antioxidant activities. Among them, dihydrochalcones are mainly found in apple leaves (Malus domestica). Glycosylated dihydrochalcones were previously found in large amounts in leaves of two genotypes of Malus with contrasting resistance to fire blight, a bacterial disease caused by Erwinia amylovora. In the present study we demonstrate that soluble polyphenol patterns comprised phloridzin alone or in combination with two additional dihydrochalcones, identified as sieboldin and trilobatin. Presence of sieboldin in young leaves correlated well with a high 1,1-diphenyl-2 picrylhydrazyl (DPPH) free radical scavenging activity. Moreover, these leaves displayed enhanced tolerance to paraquat, a photooxidative-stress generating herbicide. Interestingly, phloridzin had a high activity in the oxygen radical absorbance capacity (ORAC) assay, but its presence alone in leaves did not correlate with tolerance to paraquat. In order to further characterise the activity of these compounds, we tested their ability to prevent oxidative dependent formation of advanced glycation end-products (AGEs) and phenylephrine induced contraction of isolated rat mesenteric arteries. The antioxidant capacity of sieboldin was clearly demonstrated by showing that this compound (i) prevented vasoconstriction and (ii) inhibited AGEs formation. Both assays provided interesting information concerning a potential use of sieboldin as a therapeutic. Hence, our results strongly argue for a bioactivity of dihydrochalcones as functional antioxidants in the resistance of Malus leaves to oxidative stress. In addition, we demonstrate for the first time that sieboldin is a powerful multipotent antioxidant, effective in preventing physiopathological processes. Further work should aim at demonstrating the potential use of this compound as a therapeutic in treating free radical-involving diseases. PMID- 20022618 TI - Evaluation of a recombinant LigB protein of Leptospira interrogans serovar Canicola in an enzyme-linked immunosorbent assay for the serodiagnosis of bovine leptospirosis. AB - A recombinant leptospiral lipoprotein, LigB, was evaluated for use in the diagnosis of bovine leptospirosis by enzyme-linked immunosorbent assay (rLigB IgG ELISA). The standard reference test (Microscopic agglutination test, MAT) of 200 serum samples from cattle suspected of leptospirosis showed that 95 (47.5%) samples had positive agglutination titres, which ranged from 100 to 1600. In rLigB IgG ELISA, 49% of the samples were positive. Sensitivity of IgG ELISA for 95 bovine sera, which had MAT titres of greater than or equal to 100, were 100%. ELISA showed a specificity of 97.1% with 105 bovine sera, which were negative at a 1:50 dilution in MAT for Leptospira interrogans serovars. The results of ELISA and MAT correspond very good. When analytical specificity of IgG ELISA was evaluated using bovine serum samples from animals showing the serum antibodies to other pathogens, no cross-reaction was observed. Thus the recombinant LigB IgG ELISA can be used instead of the MAT as an aid to the diagnosis of bovine leptospirosis. PMID- 20022619 TI - [Doxycycline versus placebo trial in human prion diseases: a therapeutic challenge]. PMID- 20022620 TI - Comment on "The removal of pharmaceuticals, personal care products, endocrine disruptors and illicit drugs during wastewater treatment and its impact on the quality of receiving waters" by Barbara Kasprzyk-Hordern, Richard M. Dinsdale, and Alan J. Guwy. PMID- 20022621 TI - Detection and molecular characterization of noroviruses from five sewage treatment plants in central Italy. AB - Noroviruses (NoVs) are the most frequent etiological agents of non-bacterial gastroenteritis. These viruses are transmitted through the fecal-oral route, leading to high viral loads in sewages. The objective of this paper was to study the environmental occurrence of the most prevalent NoV strains in different wastewater treatment plants. In addition, molecular characterization of the isolated strains was performed. Two different PCR-based methods were carried out and a novel strategy was used to verify the level of RT-PCR inhibition. From May to September 2007, a total of 97 inflow and outflow samples were collected from five wastewater treatment plants in central Italy. We detected NoV by nested PCR in 96.9% of influent samples: 89.1% contained both genogroups; 4.7% contained only GI and 3.1% only GII. In effluents, we detected NoV in 78.8% of samples: 30.3% contained both genogroups, and 48.5% contained only GI. The major genotypes detected by sequencing analyses were GI/2, GI/5, GII/b, GII/4 and GII/6. This work confirms the wide circulation of NoVs in Italy with a predominance of GI strains, and the widespread distribution of NoV variants in both raw and treated wastewater. PMID- 20022622 TI - Kinetics and mechanism of the degradation of two pesticides in aqueous solutions by ozonation. AB - This study evaluated the reaction kinetics and degradation mechanism of the pesticides bromoxynil and trifluralin during conventional ozonation. The second order rate constants for the direct molecular ozone and hydroxyl radical reactions with bromoxynil and trifluralin were determined using a rapid-scan stopped-flow spectrophotometry, competition kinetics, and an organic substrate monitoring method. High reactivity toward ozone and hydroxyl radicals was observed for bromoxynil and trifluralin. The second-order rate constants for the reaction with ozone were about 10(2)M(-1)s(-1). The values of the second-order rate constants for the reaction between hydroxyl radicals and bromoxynil and trifluralin in ultrapure water at 20 degrees C were estimated to be around 8.4x10(9) and 7.5x10(9)M(-1)s(-1), respectively. The identification of oxidation by-products generated during ozonation was also addressed. It was found that hydroxylation and debromination were the primary pathways for the bromoxynil degradation, whereas hydroxylation and dealkylation were found to be the major mechanisms for trifluralin oxidation. PMID- 20022623 TI - Mass balance for POPs in hazardous and municipal solid waste incinerators. AB - The amount of different persistent organic pollutants (POPs) in the input of waste incinerators was compared to that in the output. Three cases were considered: a rotary kiln incinerating hazardous waste, a grate furnace incinerating municipal solid waste (MSW) and the same grate furnace co incinerating plastics of waste of electrical and electronic equipment (WEEE) and automotive shredder residue (ASR) with MSW. The mass balance for PCBs in the rotary kiln indicates that these POPs are destroyed effectively during incineration. The grate furnace can be a sink or source of PCDD/Fs and PCBs depending on the concentrations in the incinerated waste. In order to compare the total amount of POPs in input and output, a methodology was developed whereby the amount of POPs was weighed according to minimal risk doses (MRDs) or cancer potency factors. For both incinerators the PCDD/Fs, PCBs and polyaromatic hydrocarbons (PAHs) are the main contributors to total weighed POP output. In MSW, the PCDD/Fs, PBDD/Fs and polybrominated diphenylethers (PBDEs) are the main contributors to the weighed POP input. The ratios of the weighed POP-input over output clearly indicate that the rotary kiln incinerating hazardous waste is a weighed POP sink. The grate furnace incinerating MSW is a weighed POP sink or source depending on the POP-concentrations in the waste, but the difference between output and input is rather limited. When e.g. ASR and plastics of WEEE, containing high concentrations of PBDEs and PCBs, are co-incinerated in the grate furnace, it is clearly a weighed POP sink. PMID- 20022624 TI - Biomarker responses in Solea senegalensis exposed to sodium hypochlorite used as antifouling. AB - The time-course stress responses (0, 1, 2, and 7 d) was assessed in plasmatic, branchial and renal parameters of juveniles Solea senegalensis exposed to different concentrations of the antifouling sodium hypochlorite (0.1, 0.2, and 0.5mgL(-1)). These stress responses were only assessed for the total length of exposure (7d) at the lowest NaClO concentration due to the high toxicity of this chemical. In addition, the xenobiotic metabolism responses were evaluated by means of enzymatic activities of ethoxyresorufin O-deethylase (EROD), glutathione S-transferase (GST), glutathione reductase (GR), glutathione peroxidase (GPX), catalase (CAT), and carboxylesterase (CbE) in liver; as well as GST, GPX, CAT and acetylcholinesterase (AChE) in gill. Oxidative stress damage due to sodium hypochlorite exposure was measured by lipid peroxidation levels in liver and gill. Concentrations of 0.2 and 0.5mgL(-1) produced lethal effects after 1d and 2h of exposure, respectively. After 1d of exposure to sublethal concentration of sodium hypochlorite (0.1mgL(-1)) osmoregulatory (osmolality and chloride) and stress (cortisol, glucose and lactate) plasmatic parameters were enhanced to respect at control fish. However after 3 or 7d these parameters returned to control values. No effects were observed on plasma protein and triglyceride levels or on gill and kidney Na(+)/K(+)-ATPase activities. Diverse gill pathologies such as hypertrophy, lamellar fusion and an increase in goblet cell number and size were observed after 7d of exposure. Most biochemical parameters related to xenobiotic metabolism and oxidative stress were also significantly affected which suggests that seawater affected by sodium hypochlorite discharges from power plants, is able to alter the fish xenobiotic metabolism and generate oxidative stress. PMID- 20022625 TI - Degradation of tetracycline antibiotics: Mechanisms and kinetic studies for advanced oxidation/reduction processes. AB - This study involves elucidating the destruction mechanisms of four tetracyclines via reactions with ()OH and solvated electrons (e(aq)(-)). The first step is to evaluate the bimolecular rate constants for the reaction of ()OH and e(aq)(-). Transient absorption spectra for the intermediates formed by the reaction of ()OH were also measured over the time period of 1-250micros to assist in selecting the appropriate wavelength for the absolute bimolecular reaction rate constants. For these four compounds, tetracycline, chlortetracycline, oxytetracycline, and doxycycline, the absolute rate constants with ()OH were (6.3+/-0.1)x10(9), (5.2+/ 0.2)x10(9), (5.6+/-0.1)x10(9), and (7.6+/-0.1)x10(9) M(-1) s(-1), and for e(aq)( ) were (2.2+/-0.1)x10(10), (1.3+/-0.2)x10(10), (2.3+/-0.1)x10(10), and (2.5+/ 0.1)x10(10) M(-1) s(-1), respectively. The efficiencies for ()OH reaction with the four tetracyclines ranged from 32% to 60%. The efficiencies for e(aq)(-) reaction were 15-29% except for chlortetracycline which was significantly higher (97%) than the other tetracyclines in spite of the similar reaction rate constants for e(aq)(-) in all cases. To evaluate the use of advanced oxidation/reduction processes for the destruction of tetracyclines it is necessary to have reaction rates, reaction efficiencies and destruction mechanisms. This paper is the first step in eventually realizing the formulation of a detailed kinetic destruction model for these four tetracycline antibiotics. PMID- 20022626 TI - In vitro comparison of egg yolk-based and soybean lecithin-based extenders for cryopreservation of ram semen. AB - Substitution of egg yolk with soybean lecithin may reduce hygienic risks in extenders. Though a few studies have been performed on the effect of soybean lecithin in bull, to date evaluation of ram semen in vitro fertility after cryopreservation with use of soybean lecithin has not been studied. This study assessed the effect of 1% or 2% (wt/vol) soybean lecithin (L1 or L2) or 15% or 20% (vol/vol) egg yolk (E15 or E20) supplemented with 5% or 7% glycerol (G5 or G7) in a Tris-based medium for cryopreservation of ram (Oviss arries) semen. Although no significant difference was observed in pattern of capacitation, the best results in terms of sperm motility, viability postthaw, and cleavage rates were observed with L1G7 (51.9+/-4.8%, 48.1+/-3.5%, and 79.6+/-3.9%, respectively) and E20G7 (51.8+/-2.9%, 46.7+/-4.0%, and 72.9+/-6.4%, respectively). Our results also showed that 1% lecithin and 20% egg yolk was superior to 2% lecithin and 15% egg yolk. In terms of cleavage rate, 7% glycerol was superior to 5% glycerol. No significant difference was obtained between groups in terms of blastocysts rate per cleaved embryo. Therefore, we concluded that the optimal concentration of lecithin and egg yolk is 1% and 20%, respectively, along with 7% glycerol. In addition, our results suggest that lecithin can be used as a substitute for egg yolk. PMID- 20022627 TI - The activity of macrophage aggregates in the liver of flounder (Platichthys flesus) and wrasse (Symphodus melops) is associated with tissue damage. AB - The phagocytic activity of macrophage aggregates (MAs) in the liver of fish is characterized by high acid phosphatase activity and generation of reactive oxygen species (ROS). Question of the present study was whether these activities were associated with damage of adjacent and surrounding liver cells and the macrophages itself. Thus, the lysosomal membrane stabilities (LMS) of two different populations of lysosomes (LMS1, short destabilization periods, and LMS2, longer destabilization periods) of liver cells close to MAs of different phagocytic activity were measured as indicator for cellular integrity by computer assisted image analysis. They were then compared to the LMS of hepatocytes remote from any MA. In addition, LMS was also assessed inside the MAs. Studies were performed on adult European flounder (Platichthys flesus) and corkwing wrasse (Symphodus melops) caught at clean and polyaromatic hydrocarbon (PAH) contaminated field locations of the German Bight and the Karmoy peninsula at the West coast of Norway. In the hepatocytes adjacent to MAs which showed medium to high phagocytic activity, LMS was significantly decreased with a reduction of both, first (LMS1), and second (LMS2) destabilization periods in both species. LMS1 inside of these MAs was extremely low, whereas LMS2 was sometimes even higher compared to the liver cells. We suggest that LMS1 represents phagosomes which had contact to ROS and LMS2 represents less affected primary lysosomes. In single flounder and wrasse with liver tumors, MAs in the vicinity of these tumors showed low phagocytic activity and were not associated with adverse effects on adjacent tumor cells. Inside these MAs, LMS1 was higher compared to MAs with high phagocytic activity in wrasse from the reference site, indicating that cytotoxicity was not involved in their phagocytic deficiency. PMID- 20022628 TI - Fluorescence characteristics of chromophoric dissolved organic matter in shallow water along the Zhejiang coasts, southeast China. AB - Twenty-eight surface water samples from rivers, muddy intertidal flats, sand shores, and bedrock coasts were collected along the Zhejiang coastline in southeast China. In addition, three samples from the Changjiang (Yangtze River) were collected for comparison. CDOM (chromophoric dissolved organic matter) absorption and fluorescence excitation-emission matrix (EEM) spectroscopy, as well as nutrients and DOC were measured in these samples. According to salinity, nutrient, and DOC constituents, the 28 Zhejiang samples were categorized into four groups, i.e. highly-polluted, river derived, muddy-flat derived, and saltwater dominated ones. Among the six parameters (two humic-like and two protein-like peak intensities in fluorescence EEM contours, absorption at 300 nm, and DOC concentration) for the Zhejiang samples, any two of them were positively correlated. The submarine groundwater discharge, rather than local rivers, might have provided most of the freshwater that interacted with the saltwater during the mixing process. The high protein-like EEM peaks in samples from muddy salt marshes and rivers were probably caused by terrestrial inputs, land-based pollution, and local biological activities in combination. PMID- 20022629 TI - N-acetyl cysteine directed detoxification of 2-hydroxyethyl methacrylate by adduct formation. AB - Cytotoxicity of the dental resin monomer 2-hydroxyethyl methacrylate (HEMA) and the protective effects of N-acetyl cysteine (NAC) on monomer-induced cell damage are well demonstrated. The aim of our study was to analyze the hypothesis that the protection of NAC from HEMA cytotoxicity might be due to direct NAC adduct formation. To this end, using HPLC we first measured the actual intracellular HEMA concentrations able to cause toxic effects on 3T3-fibroblasts and then determined the decrease in intracellular and extracellular HEMA levels in the presence of NAC. In addition, by capillary electrophoresis coupled with mass spectrometry analysis (CE-MS), we evaluated NAC-HEMA adduct formation. HEMA reduced 3T3 cell vitality in a dose- and time-dependent manner. The concentration of HEMA inside the cells was 15-20 times lower than that added to the culture medium for cell treatment (0-8 mmol/L). In the presence of 10 mmol/L NAC, both intracellular and extracellular HEMA concentrations greatly decreased in conjunction with cytotoxicity. NAC-HEMA adducts were detected both in the presence and absence of cells. Our findings suggest that the in vitro detoxification ability of NAC against HEMA-induced cell damage occurs through NAC adduct formation. Moreover, we provide evidence that the actual intracellular concentration of HEMA able to cause cytotoxic effects is at least one magnitude lower than that applied extracellularly. PMID- 20022630 TI - The regulation of stem cell differentiation by cell-cell contact on micropatterned material surfaces. AB - Using the material technique recently developed by us, we prepared a micropattern on poly(ethylene glycol) (PEG) hydrogel to keep background resistant to cell adhesion for a long time, which made examination of differentiation of localized stem cells available. Our micropattern designed in this paper prevented or ensured contact between cells adhering in arginine-glycine-aspartic acid (RGD) microdomains, and thus afforded a unique way to study the effects of cell-cell contact on the lineage differentiation of stem cells while ruling out the interference of soluble factors or cell seeding concentration etc. As demonstration, mesenchymal stem cells derived from rats were examined in this study, and both osteogenic and adipogenic differentiations were found to be regulated by cell-cell contact. Isolated cells exhibited less significant differentiation than paired or aggregated cells. For those stem cells in contact, the extent of differentiation was fairly linearly related to the extent of contact characterized by coordination number. Additionally, we revealed the existence of some unknown cues besides gap junction responsible for such effects of cell-cell contact. PMID- 20022631 TI - Engineering of capillary-like structures in tissue constructs by electrochemical detachment of cells. AB - A major challenge in the development of functional thick tissues is the formation of vascular networks for oxygen and nutrient supply throughout the engineered tissue constructs. This study describes an electrochemical approach for fabrication of capillary-like structures, precisely aligned within micrometer distances, whose internal surfaces are covered with vascular endothelial cells. In this approach, an oligopeptide containing a cell adhesion domain (RGD) in the center and cysteine residues at both ends was designed. Cysteine has a thiol group that adsorbs onto a gold surface via a gold-thiolate bond. The cells attached to the gold surface via the oligopeptide were readily and noninvasively detached by applying a negative electrical potential and cleaving the gold thiolate bond. This approach was applicable not only for a flat surface but also for various configurations, including cylindrical structures. By applying this approach to thin gold rods aligned in a spatially controlled manner in a perfusion culture device, human umbilical vein endothelial cells (HUVECs) were transferred onto the internal surface of capillary structures in collagen gel. In the subsequent perfusion culture, the HUVECs grew into the collagen gel and formed luminal structures, thereby forming vascular networks in vitro. PMID- 20022633 TI - Nuclear calcium controls the apoptotic-like cell death induced by d-erythro sphinganine in tobacco cells. AB - Studies performed in animals have highlighted the major role of sphingolipids in regulating the balance between cell proliferation and cell death. Sphingolipids have also been shown to induce cell death in plants via calcium-based signalling pathways but the contribution of free cytosolic and/or nuclear calcium in the overall process has never been evaluated. Here, we show that increase in tobacco BY-2 cells of the endogenous content of Long Chain Bases (LCBs) caused by external application of d-erythro-sphinganine (DHS) is followed by immediate dose dependent elevations of cellular free calcium concentration within the first minute in the cytosol and 10min later in the nucleus. Cells challenged with DHS enter a death process through apoptotic-like mechanisms. Lanthanum chloride, a general blocker of calcium entry, suppresses the cellular calcium variations and the PCD induced by DHS. Interestingly, dl-2-amino-5-phosphopentanoic acid (AP5) and [(+)-dizocilpine] (MK801), two inhibitors of animal and plant ionotropic glutamate receptors, suppress DHS-induced cell death symptoms by selectively inhibiting the variations of nuclear calcium concentration. The selective action of these compounds demonstrates the crucial role of nuclear calcium signature in controlling DHS-induced cell death in tobacco cells. PMID- 20022632 TI - Regulation of osteogenic differentiation of rat bone marrow stromal cells on 2D nanorod substrates. AB - Bone marrow stromal cells (BMSCs) possess multi-lineage differentiation potential and can be induced to undergo differentiation into various cell types with the correct combination of chemical and environmental factors. Although, they have shown great prospects in therapeutic and medical applications, less is known about their behavior on nanosurfaces mimicking the extra cellular matrix (ECM). In this report we have employed 2D substrates coated with tobacco mosaic virus (TMV) nanorods to study the differentiation process of BMSCs into osteoblast like cells. TMV is a rod-shaped plant virus with an average length of 300 nm and diameter of 18 nm. The osteogenic differentiation of BMSCs on TMV was studied over time points of 7, 14 and 21 days. We examined the temporal gene expression changes during these time points by real-time quantitative PCR (RT-qPCR) analysis. As expected, osteo-specific genes (osteocalcin, osteopontin and osteonectin) were upregulated and showed a maximum change in expression on TMV at 14 days which was 7 days earlier than on tissue culture plastic (TCP). Based on the genes expression profile generated by RT-qPCR experiments, we proposed that the early interaction of cells with TMV triggers on signaling pathways which regulate speedy expression of osteocalcin in turn, resulting in early mineralization of the cells. To further investigate these regulating factors we studied global changes in gene expression (DNA microarray analyses) during osteogenic differentiation on the nanosubstrate. Multitudes of genes were affected by culturing cells on nanorod substrate, which corroborated our initial PCR findings. Microarray analysis further revealed additional targets influenced by the presence of nanorods on the surface, of which, the expression of bone morphogenetic protein 2 (BMP2) was of particular interests. Further investigation into the temporal change of BMP2, revealed that it acts as a major promoter in signaling the early regulation of osteocalcin on TMV coated substrates. PMID- 20022634 TI - Hotspot mutations of PIK3CA and AKT1 genes are absent in multiple myeloma. AB - The phosphatidylinositol-3-kinases PI3K/AKT pathway regulates many growth and survival mechanisms in the cell, and it has been implicated in development and progression of many human cancers, including multiple myeloma. Recently, many reports have revealed that the PIK3CA gene which encodes the p110 catalytic subunit of PI3K kinase is mutated in many human malignancies. To investigate the oncogenic role of PI3K/AKT pathway in multiple myeloma we sequenced three hot exons: exons 9 and 20 of PIK3CA gene and exon 3 of AKT1 gene in 27 multiple myeloma patients. Our results indicate the absence of the four hot spot mutations E542K, E545K, H1047R and E17K in all studied cases. These findings suggest that PI3K/AKT mutations may not play a major role in multiple myeloma. PMID- 20022635 TI - Muscular cholinesterase activities and lipid peroxidation levels as biomarkers in several Mediterranean marine fish species and their relationship with ecological variables. AB - Muscular cholinesterase activities, as potential markers of neurotoxic exposure, and lipid peroxidation levels, indicative of oxidative stress damage, both currently used in early-warning pollution monitoring, were characterised in eighteen fish species of ecologic and/or economic importance. These species comprise five orders and eleven families of teleosts and two species of elasmobranchs, feed using different strategies (benthic, epibenthic, endobenthic and pelagic), belong to different trophic levels and express different swimming behaviour. Their habitat ranges from 50 to 60 m (shallow or continental shelf) and 600 to 850 m (middle continental slope). Sampling took place in front of the Barcelona coast (NW Mediterranean) during four seasonal cruises in 2007. In the summer sampling, another site potentially exposed to a different pollution load (Vilanova) was included for comparison. Species, seasonal and site differences were tested and discussed in relation to chemical analysis of the local sediment, systematic position, habitat depth, feeding strategy, trophic level and swimming activity. Greater inter species differences rather than seasonal or site trends were seen in accordance to little pollution fluctuations. Higher cholinesterase activities were recorded in suprabenthos feeders, regardless of depth habitat, whereas LP levels were similar in all species except for the shark Scyliorhinus canicula in which they were consistently elevated. This study confirms and broadens former observations carried out with a more reduced number of fish species (Sole et al., 2008a). PMID- 20022636 TI - Isolation and immunological characterization of a novel Cladosporium herbarum allergen structurally homologous to the alpha/beta hydrolase fold superfamily. AB - Because the ascomycete Cladosporium herbarum embodies one of the most important, world-wide occurring fungal species responsible for eliciting typical IgE mediated hypersensitivity reactions ranging from rhinitis and ocular symptoms to severe involvement of the lower respiratory tract, a more comprehensive definition of its detailed allergen repertoire is unquestionably of critical medical as well as therapeutic significance. By screening a C. herbarum cDNA library with IgE antibodies pooled from 3 mold-reactive sera, we were able to identify, clone and affinity-purify a novel allergen candidate (29.9 kDa) exhibiting considerable (three-dimensional) homology to the alpha/beta hydrolase fold superfamily. The latter covers a collection of hydrolytic enzymes of widely differing phylogenetic origin as well as catalytic activity (operating in countless biological contexts) that in general exhibit only little sequence similarity yet show a remarkable conservation of structural topology. Our present study (i) characterizes recombinant non-fusion C. herbarum hydrolase as a natively folded, minor mold allergen that displays a prevalence of IgE reactivity of approximately 17% in our in vitro immunoblot experiments, (ii) proposes the existence of several putative (speculatively cross-reactive) ascomycete orthologues as determined via genome-wide in silico predictions, and (iii) finally implies that C. herbarum hydrolase could be included in forthcoming minimal testing sets when fungal allergy is suspected. PMID- 20022637 TI - Ocular and orbital manifestations of the inherited bone marrow failure syndromes: Fanconi anemia and dyskeratosis congenita. AB - PURPOSE: The inherited bone marrow failure syndromes (IBMFS) are a heterogeneous group of genetic disorders that share the inability of the bone marrow to produce an adequate number of blood cells. The 4 most frequent syndromes are Fanconi anemia (FA), dyskeratosis congenita (DC), Diamond-Blackfan anemia (DBA), and Shwachman-Diamond syndrome (SDS). All 4 syndromes have been associated with various physical abnormalities. As part of a genotype/phenotype/cancer susceptibility study, we determined the prevalence of ophthalmic manifestations in these 4 syndromes. DESIGN: Cross-sectional study of a patient cohort. PARTICIPANTS: Seventy-five patients with an IBMFS and 121 of their first-degree relatives were seen in the National Eye Institute, National Institutes of Health, from 2001 to 2007. The patient group included 22 with FA, 28 with DC, 19 with DBA, and 6 with SDS. METHODS: Every participant underwent a complete ophthalmic evaluation and digital facial photography with an adhesive paper ruler on the patient's forehead for an internal measure of scale. Interpupillary distance (IPD), inner canthal distance (ICD), outer canthal distance (OCD), palpebral fissure length (PFL), and corneal diameter (CD) were measured. Thirteen of the 22 patients with FA underwent axial length (AL) measurements by A-scan ultrasonography. MAIN OUTCOME MEASURES: Type and prevalence of ophthalmic manifestations. RESULTS: Ninety-five percent of patients with FA had at least 1 abnormal parameter, and 25% of patients had at least 4 abnormal parameters. Eighty-two percent of patients had small palpebral fissures, 69% of patients had simple microphthalmia, 64% of patients had small OCD, 55% of patients had microcornea, 28% of patients had ptosis, and 6% of patients had epicanthal folds. In patients with DC, abnormalities of the lacrimal drainage system (29%) were the most prevalent findings, followed by retinal abnormalities (pigmentary changes, retinal neovascularization, retinal detachment, exudative retinopathy) in 21%, cicatricial entropion with trichiasis and blepharitis in 7% each, and sparse eyelashes and congenital cataract in 3.5% each. No significant ophthalmic abnormalities were seen in patients with DBA or SDS. CONCLUSIONS: Syndrome specific ocular findings are associated with FA and DC and may antedate diagnosis of the specific syndrome. Early recognition of these abnormalities is important for optimal management. PMID- 20022639 TI - Primary orbital liposarcoma. AB - PURPOSE: To describe 6 new cases of primary orbital liposarcoma and provide a review of the relevant literature. DESIGN: Noncomparative consecutive case series and literature review. PARTICIPANTS: Six patients with primary orbital liposarcoma. METHODS: Review of patient charts, imaging, and histopathology; literature review. MAIN OUTCOME MEASURES: Patient demographics; clinical presentations; results of radiologic imaging; histopathology; surgical techniques used and their complications; other treatment modalities; outcomes and recurrences. RESULTS: Six cases of primary orbital liposarcoma were identified, 5 of which were primary presentations and 1 of which was a recurrence. In 4 cases, exenteration was deferred, resulting in recurrence of disease in all 4. All cases were exenterated, and 2 cases had local recurrence despite exenteration. Two cases were associated with the Li-Fraumeni syndrome and other malignancies. Literature review identified 34 other cases of primary orbital liposarcoma, which, partly because of its rarity, is frequently initially misdiagnosed. The most common subtype is myxoid (56.8%); other types are pleomorphic (10.8%) and well differentiated (29.7%). Well-differentiated tumors have the best prognosis. Non-exenterating surgery was associated with recurrence, although recurrence post exenteration also occurred. Although radiotherapy has an established role in the treatment of nonorbital liposarcoma, the role of both radiotherapy and chemotherapy in the management of primary orbital liposarcoma is still unclear. CONCLUSIONS: Orbital liposarcoma remains a diagnostic and surgical challenge. Exenteration remains the treatment of choice, but clinicians must also be aware that liposarcoma may herald the diagnosis of the Li-Fraumeni familial cancer syndrome. PMID- 20022638 TI - The complement component 5 gene and age-related macular degeneration. AB - OBJECTIVE: To investigate the association between variants in the complement component 5 (C5) gene and age-related macular degeneration (AMD). DESIGN: Separate and combined data from 3 large AMD case-control studies and a prospective population-based study (The Rotterdam Study). PARTICIPANTS: A total of 2599 AMD cases and 3458 ethnically matched controls. METHODS: Fifteen single nucleotide polymorphisms (SNPs) spanning the C5 gene were initially genotyped in 375 cases and 199 controls from The Netherlands (The Amsterdam/Rotterdam Netherlands [AMRO-NL] study population). Replication testing of selected SNPs was performed in the Rotterdam Study (NL) and study populations from Southampton, United Kingdom (UK), and New York, United States (US). MAIN OUTCOME MEASURES: Early and late stages of prevalent and incident AMD, graded according to (a modification of) the international grading and classification system of AMD. RESULTS: Significant allelic or genotypic associations between 8 C5 SNPs and AMD were found in the AMRO-NL study and this risk seemed to be independent of CFH Y402H, LOC387715 A69S, age, and gender. None of these findings could be confirmed consistently in 3 replication populations. CONCLUSIONS: Although the complement pathway, including C5, plays a crucial role in AMD, and the C5 protein is present in drusen, no consistent significant associations between C5 SNPs and AMD were found in any of these studies. The implications for genetic screening of AMD are discussed. PMID- 20022640 TI - Pituitary volume in patients with bipolar disorder and their first-degree relatives. AB - BACKGROUND: Hypothalamic-pituitary-adrenal (HPA) axis dysregulation has been reported in bipolar disorder (BD), but previous magnetic resonance imaging (MRI) studies of pituitary gland volume in BD have yielded inconsistent findings. In addition, the contribution of genetic factors to the pituitary changes in BD remains largely unknown. METHOD: We used MRI to investigate the pituitary volume in 29 remitted patients with BD, 49 of their first-degree relatives (of whom 15 had a diagnosis of Major Depressive Disorder), and 52 age- and gender-matched healthy controls. RESULTS: BD patients had a significantly larger pituitary volume compared with their relatives and healthy controls. Pituitary volume did not differ between controls and healthy relatives or relatives diagnosed with major depression. LIMITATIONS: Direct measures of HPA function (i.e., hormonal levels) were not available. CONCLUSIONS: These findings suggest that enlarged pituitary volume is associated with disease expression but not genetic susceptibility to BD. PMID- 20022641 TI - Prevalence of DFNB1 mutations in Argentinean children with non-syndromic deafness. Report of a novel mutation in GJB2. AB - OBJECTIVE: Mutations in DFNB1 locus, containing GJB2 (connexin 26) and GJB6 (connexin 30) genes, are the most common cause of autosomal recessive non syndromic hearing loss. More than 100 mutations in GJB2 have been reported worldwide. Two deletions in GJB6, del(GJB6-D13S1830) and del(GJB6-D13S1854), have been found to be frequent in the Spanish population. The aim of this study was to determine the prevalence of GJB2 mutations and both GJB6 deletions in Argentinean children with non-syndromic deafness. METHODS: This study included 94 unrelated children with moderate to profound non-syndromic sensorineural hearing impairment. Molecular analysis was performed using a tiered approach. All DNA samples were screened for c.35delG mutation by PCR/RFLP. Samples from patients who were not homozygous for c.35delG were analysed for the presence of GJB6 deletions by PCR multiplex. The samples that remained unresolved after screening were further analysed by direct sequencing of GJB2 coding region. Finally, the splice site mutation IVS1+1G-->A was analysed by PCR/RFLP. RESULTS: Sequence variations in the GJB2 and GJB6 genes were found in 49 of the 94 unrelated patients. The most prevalent GJB2 mutation, c.35delG, was found in 40 of the 68 pathogenic alleles with the second most common allele being p.R143W (4/68). Fourteen sequence variations other than c.35delG were identified. Seven already described mutations were present in more than one allele; among them, IVS1+1G- >A, the rare splice site mutation flanking exon 1. In addition to known disease related alterations, a novel GJB2 mutation, c.262G>C (p.A88P), was also identified. Six alleles were identified carrying GJB6 deletions; the most prevalent was del(GJB6-D13S1830). The frequency of the latter was found to be as high as that found in Spain from where Argentina has received one of its major immigration waves. CONCLUSIONS: The overall frequency of GJB2/GJB6 mutations in the present sample is in agreement with other Caucasian populations. As expected, c.35delG was the most prevalent mutation. The deletion del(GJB6-D13S1830) was the second most common mutation. These findings reinforce the importance of the study of GJB2/GJB6 genes in diagnosis to provide early treatment and genetic counselling. PMID- 20022642 TI - Comparison of six sewage effluents treated with different treatment technologies- population level responses in the harpacticoid copepod Nitocra spinipes. AB - Since conventional treatment technologies may fail in removing many micro pollutants, there is currently a focus on the potential of additional treatment technologies for improved sewage treatment. The aim of the present study was to evaluate six different effluents from Henriksdal Sewage Treatment Plant in Stockholm, Sweden. The effluents were; conventionally treated effluent (chemical phosphorous removal in combination with an activated sludge process, including biological nitrogen removal and a sand filter), with additional treatments individually added to the conventional treatment; active carbon filtration, ozonation at 5 mg l(-1), ozonation at 15 mg l(-1), ozonation at 5 mg l(-1)+moving bed biofilm reactor and irradiation with ultraviolet radiation+hydrogen peroxide. The evaluation was done by characterizing and comparing the effluents using a Lefkovitch matrix model based on a life cycle test with the harpacticoid copepod Nitocra spinipes, combined with analysis of juvenile development and survival over time. The conventionally treated effluent resulted in the most negative effects, leading to the conclusion that all additional treatments in the present study created effluents with less negative impacts on the copepod populations. The ozone treatments with the low dose treatment in particular, resulted in the overall least negative effects. Moving bed biofilm reactor combined with ozone did not improve the quality of the effluent in the sense that slightly more negative effects on the population abundance were seen for this treatment technology compared to ozonation alone. The active carbon treatment had more negative effects than the ozone treatments, most of which could possibly be explained by removal of essential metal ions. The effluent which was treated with ultraviolet radiation+hydrogen peroxide resulted in few developmental and survival effects over time, but still showed negative effects on the population level. Matrix population modeling proved a useful tool for biologically characterizing and comparing the effluents. Basing the assessment either on the individual level data (development and survival over time or total reproductive output) or the population level data (lambda values and projected population abundances) would not have resulted in the same conclusions as combining both analyses. The juvenile development and survival over time allowed for closer monitoring of the important molting process, whereas the population modeling provided an integrated measure of potential effects at the population level. If the dilution of the effluent in the recipient is considered, the biological effects recorded in the present study were not of substantial significance for the copepod populations, regardless of treatment technology. PMID- 20022643 TI - An unusual case of cor triatriatum sinister presenting as pulmonary oedema during labor. AB - A 35 years old second gravida was admitted to our department with acute pulmonary oedema after labor. Echocardiography disclosed an intra-atrial membrane prolapsing through the mitral valve in diastole, with two small orifices creating a mean pressure gradient of 7.9 mm Hg. However the membrane clearly originated away from the mitral valve ring, proximal to the left atrial appendage, consistent with cor triatriatum sinister. Cor triatriatum sinister rarely presents in adulthood and although forward movement of the membrane towards the mitral valve funnel has been described, such a degree of prolapse through the valve has never been reported. PMID- 20022644 TI - Pheochromocytoma-induced ventricular tachycardia and reversible cardiomyopathy. AB - A case of a 48-year-old male is described with recurrent paroxysmal ventricular tachycardia originating near the region of the posterior fascicle, as an initial presentation of a primary norepinephrine producing pheochromocytoma, including reversible catecholamine-induced cardiomyopathy. The ventricular arrhythmia was terminated by procainamide and labetalol. After adrenalectomy no recurrent ventricular arrhythmia was observed. PMID- 20022645 TI - Plasma CA-125 level is related to both sides of the heart: a retrospective analysis. AB - OBJECTIVE: CA-125 is an old marker, recently shown to be associated with systolic heart failure. We aimed to search for factors influencing its plasma level. DESIGN: 430 patients with one available CA-125 level were investigated retrospectively. 150 patients who had echocardiographic records were enrolled into final analysis. Patients were followed up, hospitalization and mortality were noted. RESULTS: CA-125 levels were negatively correlated with ejection fraction (r=-0.269, p=0.001) and positively correlated with systolic pulmonary artery pressure (r=0.370, p<0.001). In the whole group, patients with right ventricular dilatation (n=68) had significantly higher CA-125 levels compared to those without right ventricular dilatation (n=82) (125.8+/-118.4 U/ml vs.16.9+/ 16.5 U/ml, p<0.001). Presence of depressed ejection fraction (B=1.837, p=0.004), presence of right ventricular dilatation (B=4.294, p=0.002) and presence of pericardial effusion (B=1.913, p=0.018) were independent predictors of high CA 125 levels. After follow up, patients with high CA-125 level encountered more frequent hospitalization and mortality, and atrial fibrillation was more frequent among those with high CA-125. CONCLUSION: Our data suggests that plasma levels of CA-125 seem to be determined by left ventricular ejection fraction, right ventricular dilatation and presence of pericardial effusion in a group of all comers. It seems prudent to consider these factors before integrating CA-125 into clinical practice. PMID- 20022646 TI - Assessing the similarity of antimicrobial resistance phenotypes among fecal Escherichia coli isolates from two aggregated occupational cohorts of humans versus swine using cluster analysis and multivariate statistics. AB - Statistical methods employed to analyze antimicrobial resistance (AR) phenotypic data have largely focused on multiple individual antimicrobial resistance outcomes without considering the pharmacologic and biological dependence among these data. In our 3-year longitudinal study, the relationship between AR phenotype of E. coli isolates from integrated multi-site group cohorts of humans and swine and the following risk factors: host-species (human versus swine) and human vocation (swine-workers versus non-workers) was assessed; first, by using cluster analysis techniques and then multivariate generalized estimating equation (GEE) models. Human sewage wastewater draining from occupation-specific housing and swine fecal E. coli isolates (n=3,113 and 3,428, respectively) were tested for antimicrobial susceptibility to 15 agents on the 2003 NARMS panel using the Sensititre system (Trek Diagnostics, Cleveland, OH). The MIC values for each isolate were interpreted according to standardized breakpoints into resistant or susceptible. The phenotypic data (n=6,541) were cluster-analyzed using Ward's minimum variance with Jaccard's distance measure. The multivariate relationships of E. coli cluster membership with the risk factors in the study were assessed using a multivariate GEE model in a SAS((R)) macro to adjust for the multiple cluster dependencies as well as adjusting for response dependencies within each unit location. The cluster solution that best described our entire dataset and where the multivariate GEE model converged was 14. In general, the adjusted odds ratios of the multiple clusters (i.e., 14 clusters) for human isolates were significantly (P<0.05) at a higher odds of being in the pansusceptible cluster (OR=12.8), and also in clusters that contained high levels of resistance to amoxicillin/clavulanic acid, ampicillin, cefoxitin, nalidixic acid, sulfisoxazole, and/or trimethoprim/sulfamethoxazole, when compared to swine isolates. The adjusted odds-ratios of the multiple clusters for non-swine worker isolates were at significantly (P<0.05) higher risk of being in the pansusceptible cluster (OR=13.6) compared to swine-worker isolates (OR=12.1) (swine isolates were the referent group). In general, the adjusted odds-ratios of the multiple clusters for swine-worker E. coli isolates were significantly (P<0.05) at higher odds of being in multi-resistant clusters (defined as resistant to >or=3 antimicrobial agents) as compared to non-swine worker isolates. Considering vocation, swine-worker E. coli isolates exhibited increased odds of falling in multi-drug resistance clusters compared to those isolates arising from non-swine-workers. PMID- 20022647 TI - Attitudes towards the Canadian quality milk program and use of good production practices among Canadian dairy producers. AB - To harmonize good production practices (GPP) for dairy producers in Canada, the Canadian dairy industry has developed and is implementing a program called Canadian Quality Milk (CQM). A postal questionnaire was administered to all Canadian dairy producers enrolled in dairy herd-improvement organizations in 2008 (n=10,474) to investigate their attitudes towards the program and to establish baseline information on their use of GPP. The response percentage was 20.9% (2185/10,474). Two-thirds of producers (67.6%) reported participation in CQM and 61.4% of these indicated that the requirements were easy to implement. Most producers (85.0%) reported the use of cats as a pest-control method in their barns. For dead-livestock disposal, 65.0% and 38.0% indicated use of a collection service and burial, respectively. Nearly 40.0% of respondents indicated that they purchase replacement cattle, and somatic cell-count score was the main health indicator considered before purchase. Over 70% of producers reported that they clean and disinfect maternity, calf and weaned-calf pens, while only 34.1% and 53.1% reported that they provide visitors and employees, respectively, with clean clothes and boots. Through latent-class analysis, five groups (classes) of producers with distinctive patterns of reported use of GPP were identified. These were labelled as "minimal", "sanitation-only", "employee-visitor hygiene", "typical" and "ideal" user groups, with 11.1%, 23.8%, 20.2%, 37.1% and 7.7% of respondents, respectively. Respondents in the "ideal users" group had a higher probability of reporting the use of each GPP and were more likely to have completed an educational course in food safety compared to respondents in each other group. They were also more likely to have a herd size in the uppermost quartile (>65 cows) and report participation in CQM compared to each other group except the "employee-visitor hygiene users". The greatest differences were observed when compared to the "minimal users" group for completion of a food safety course (OR=2.81), participation in CQM (OR=2.39) and having a herd size of >65 vs. <36 cows (OR=3.04). Targeted education of dairy producers on the importance of various GPP (e.g. detailed health assessments of replacement cattle before purchase) for infection control is warranted. PMID- 20022648 TI - Modelling collective effectiveness of voluntary vaccination with and without incentives. AB - Control strategies of infectious diseases at a regional level can rely on vaccination. When the decision to vaccinate is made by each farmer on a voluntary basis, a farmer vaccinates to protect his own herd. If the vaccinated herds are protected against infection, the risk for other herds is reduced, creating a positive externality (i.e. an impact on a third party not directly involved in the decision). The objectives of this study are to evaluate the effectiveness at a regional level of voluntary vaccination and to analyse the effect of financial incentives and compensation for losses due to disease considering the externalities due to vaccination. We developed a dynamic deterministic model, based on a decision model interacting with an epidemiological state transition model. It determines the proportion of farmers who vaccinate over time, and the evolution of the prevalence of infected herds in a population of herds exposed to the same risk. The behaviour of farmers is modelled assuming perfect information (farmers are fully informed about prevalence, costs of disease and costs of vaccination). The decision to vaccinate is made according to a rational economic behaviour: farmers make their choices to maximize their expected utility, considering their risk aversion. The expected utility is calculated with a decision tree. The epidemiological model is based on a SIR (Susceptible, Infectious, Recovered) model modified to consider imperfect vaccination that protects herds for one year. The model is studied analytically and by simulation. For simulations, we define the epidemiological parameters of the model (transmission rate and average duration of the infectious and recovered states), the herd level effectiveness of the vaccination, the cost of vaccination, the reduction of income due to the disease, the incomes of the farmers and the risk aversion of the farmers. These parameters are based on an endemic disease, BVD (Bovine Viral Diarrhoea). The model shows that voluntary vaccination cannot eradicate the modelled disease. Risk aversion of farmers leads to a lower prevalence at the equilibrium than when farmers are risk neutral. Incentives for vaccination decrease the prevalence but do not result in eradication of the disease. Compensating for disease losses in newly infected farms leads to an increase in prevalence, due to moral hazard (farmers behave less carefully than if they were fully exposed to the risk), except if it is restricted to vaccinated herds. PMID- 20022649 TI - Effects of chitosan films on the growth of Listeria monocytogenes, Staphylococcus aureus and Salmonella spp. in laboratory media and in fish soup. AB - The objective of this study was to assess the antimicrobial effectiveness of chitosonium acetate films on the growth of Listeria monocytogenes, Salmonella spp. and Staphylococcus aureus. The samples were tested in both laboratory conditions using Tryptone Soy Broth (TSB) and in a real food system using fish soup. The study was carried out at different temperatures (4, 12, and 37 degrees C) in order to discern the influence of such variables. Moreover, a sensory evaluation of the final product was performed as a parameter of consumer acceptance. The results showed a significant reduction of the bacterial growth, which greatly depended on the bacteria type, the temperature of incubation and the food substrate. Although the effectiveness of chitosan films decreased in the fish soup, neither the sensory properties nor the pH of the soup was affected upon their addition. The application of chitosonium acetate as an internal coating of the packaging material could be a very suitable means to assure safety of liquid food products such as fish soup at the range of temperatures studied. PMID- 20022650 TI - Reducing oyster-associated bacteria levels using supercritical fluid CO2 as an agent of warm pasteurization. AB - An innovative approach to Post-Harvest Processing (PHP) of oysters is introduced focusing on the effects of supercritical carbon dioxide (scCO(2)) on bacterial contaminants trapped in the digestive system of oysters. Oysters were exposed to scCO(2) under two conditions: (1) 100 bar and 37 degrees C for 30 min and (2) 172 bar and 60 degrees C for 60 min. Using FDA standard guidelines for food analysis, variations in the Aerobic Plate Count (APC) were assessed. It was established that exposing oysters to CO(2) at 100 bar and 37 degrees C for 30 min and at 172 bar and 60 degrees C for 60 min induced 2-log and 3-log reductions in the APC respectively. The decrease in the microbial load as a result of treatment with scCO(2) was found to be significant (P=0.002). A release of adductor muscles from the shell was noted in oysters treated at 172 bar and 60 degrees C for 60 min; this was not the case for oysters treated at 100 bar and 37 degrees C for 30 min. A blind study allowing sensory analysis of treated vs. untreated oysters was also completed and no significant change in the physical appearance, smell, or texture was recorded. In this paper, we also report the effect of scCO(2) on several bacterial isolates, including a referenced ATCC strain of a non-pathogenic Vibrio (Vibrio fischeri) as well as several other bacterial isolates cultured from oyster' tissues and found to share biochemical features common to pathogenic Vibrio strains. A complete inactivation (minimum 7-log reduction) was achieved with these latter bacterial isolates. A 6-log reduction was observed with V. fischeri. PMID- 20022651 TI - Development and validation of a risk-score model for subjects with impaired glucose tolerance for the assessment of the risk of type 2 diabetes mellitus-The STOP-NIDDM risk-score. AB - AIMS: To develop a risk-score model, based on available clinical data to assess absolute risk of type 2 diabetes among people with impaired glucose tolerance (IGT). METHODS: Data from the study to prevent non-insulin dependent diabetes mellitus (STOP-NIDDM) investigating acarbose treatment in individuals with IGT were used to develop multivariable Cox proportional hazards model for the time to onset of diabetes. The final model equation was externally validated using data from the Finnish Cardiovascular Risk Factor (FINRISK) population. RESULTS: The risk-score model included the variables acarbose treatment, gender, serum triglyceride level, waist circumference, fasting plasma glucose, height, history of cardiovascular disease (CVD) and hypertension. The final model yielded an area under the receiver-operating-characteristic curve (AUC(ROC)) of 0.64 when applied to people with IGT in the STOP-NIDDM, and 0.84 and 0.90 when applied to FINRISK population with IGT alone and IGT and normal glucose tolerance combined, respectively; AUC(ROC) is a measure of the discriminatory power of the model (1, perfect discrimination). CONCLUSIONS: The STOP-NIDDM risk-score is a simple and validated tool that can identify high-risk individuals with IGT who would benefit most from type 2 diabetes or CVD prevention strategies, such as lifestyle management or early acarbose treatment. PMID- 20022652 TI - Identification of two novel variants in PRKAG2 gene in Tunisian type 2 diabetic patients with family history of cardiovascular disease. AB - We report the identification of two novel polymorphisms in the PRKAG2 gene and preliminary association study between 5'-UTR and exon 1 polymorphisms with susceptibility to type 2 diabetes. No association with type 2 diabetes was identified. However, one of these newly identified polymorphisms (p.Ser20Ile) is likely associated with cardiac disease. PMID- 20022653 TI - Vital sign triage to rule out diabetic ketoacidosis and non-ketotic hyperosmolar syndrome in hyperglycemic patients. AB - AIMS: To develop a prediction algorithm to rule out diabetic ketoacidosis (DKA) and non-ketotic hyperosmolar syndrome (NKHS) based on vital signs for early triage of patients with diabetes. METHODS: The subjects were consecutive adult diabetic patients with hyperglycemia (blood glucose >or=250mg/dl) who presented at an emergency department. Based on a derivation sample (n=392, 70% of 544 patients at a hospital in Okinawa), recursive partitioning analysis was used to develop a tree-based algorithm. Validation was conducted using the other 30% of the patients in Okinawa (n=152, internal validation) and patients at a hospital in Tokyo (n=95, external validation). RESULTS: Three risk groups for DKA/NKHS were identified: a high-risk group of patients with glucose >400mg/dl or systolic blood pressure <100mmHg; a low risk group of patients with glucose or=100mmHg, pulse 150s. The OR for trend was 0.41, 95% CI=0.25-0.71; P=0.001. The association between shock timing and survival was significant for patients with more than 150s shock delay (OR=0.19; 95% CI=0.04-0.71; P=0.02) or for trend in shock timing (0.42, 95% CI=0.20-0.84; P=0.02) after multivariable adjustment for prognostic factors age and slope of ventricular fibrillation. CONCLUSIONS: ALS takeover delays the next shock delivery in almost two-third of cases. This delay is associated with decreased survival. PMID- 20022684 TI - Pregnancy in immigrants: are we doing the best we can? PMID- 20022685 TI - IVF outcome in women with PCOS, PCO and normal ovarian morphology. AB - OBJECTIVE: To examine the outcome of IVF in women who have normal ovaries, ovulatory PCO or PCOS. STUDY DESIGN: Analysis of a prospectively collected database in an assisted conception unit in a university teaching hospital including 290 women <37 years of age undergoing their first IVF cycle. The main outcome measure was severe OHSS requiring hospitalization. RESULTS: Severe OHSS rates were significantly higher in women with PCO (12.6%) and PCOS (15.4%) compared to those with normal ovaries (2.7%). Coasting was used significantly more often. Live birth rates per cycle started are similar among women with PCO (38%), PCOS (37%) and normal ovaries (40%). CONCLUSION: Women with ovaries of polycystic morphology are at increased risk of developing severe OHSS and of requiring avoidance techniques such as coasting, regardless of ovulatory status. However, live birth rates per cycle are similar to women with normal ovaries. PMID- 20022686 TI - Hormones and mood across the first postpartum year. PMID- 20022687 TI - Conformational substates of ferricytochrome c revealed by combined optical absorption and electronic circular dichroism spectroscopy at cryogenic temperature. AB - We have investigated the heterogeneity of the Fe(III)-Met80 linkage of horse heart ferricytochrome c by probing the 695nm charge transfer band with absorption and electronic circular dichroism (ECD) spectroscopy. In order to verify the connection between conformational substates of the Fe(III)-Met80 linkage and the 695nm band spectral heterogeneity, we have performed experiments as a function of pH (neutral and acidic) and temperature (room and 20K). At room temperature, the ECD spectrum is blue shifted with respect to the absorption one; the shift is more pronounced at acidic pH and is compatible with the presence of sub-bands. ECD measurements at 20K highlighted the heterogeneous nature of the 695nm band and provided direct experimental evidence for the presence of sub-bands. Indeed, while the absorption spectra remained deceivingly unstructured, the ECD spectra showed well resolved peaks and shoulders. A consistent fit of the 20K absorption and ECD spectra showed that five Gaussians (each centered at the same frequency in the absorption and ECD spectrum) are able to reproduce the observed lineshapes. A careful analysis of frequency shifts and intensity ratios of these sub-bands enabled us to identify at least three distinct sub-bands arising from taxonomic conformational substates of the Fe(III)-Met80 linkage. In view of the major influence of the Fe(III)-Met80 linkage on the redox potential of ferricytochrome c, we speculate that these spectrally distinguishable substates may have different functional roles. PMID- 20022689 TI - Effects of subthalamic nucleus deep brain stimulation on sweating function in Parkinson's disease. AB - PURPOSE: To assess the impact of subthalamic nucleus (STN) deep brain stimulation (DBS) on the sweating function in patients with advanced Parkinson's disease (PD). METHODS: Nineteen patients with idiopathic PD (mean age+/-SD, 61.58+/-9.47) were examined immediately before and 6 months after DBS. Each examination session included registration of autonomic symptoms by means of a semi-structural questionnaire and recording of sympathetic skin response (SSR) from both palms and one sole. The neurophysiological measurements were compared to those of 19 matched for sex and age healthy controls. RESULTS: Six months post-DBS motor improvement was amounted to 65.9% and the daily levodopa equivalent dose was decreased by 36.4%. Post-operatively, dyshidrosis manifestations were reduced by 66.7% (pre-DBS sudomotor dysfunction in 47.4% of patients and sudomotor fluctuation in 57.1% of the above patients). There were no significant differences in-between pre- and post-DBS results of SSR study. However, the number of patients with at least one abnormal SSR pre-operative was reduced from 6 to 3 post-operative. No correlation was found between this neurophysiological finding and the change of clinical symptoms of hyperhidrosis or the DBS motor improvement. CONCLUSIONS: These results, although based on a small sample, suggest that STN DBS, in addition to the effect to the mobility, might also favorably regulate sweat in idiopathic PD. PMID- 20022690 TI - HA14-1 sensitizes TNF-alpha-induced apoptosis via inhibition of the NF-kappaB signaling pathway: involvement of reactive oxygen species and JNK. AB - Nuclear factor-kappa B (NF-kappaB) activation by tumor necrosis factor-alpha (TNF alpha) attenuates the TNF-alpha-induced apoptosis pathway. Thus, blockage of NF kappaB activity may improve the anti-cancer activity of TNF-alpha. HA14-1 induces apoptosis in various human cancer cells, and the molecular mechanisms of this action remain to be fully characterized. The present study evaluated the involvement of NF-kappaB, reactive oxygen species (ROS), and c-Jun N-terminal kinase (JNK) in the effects of HA14-1 by examining the sensitization effect on TNF-alpha-induced apoptosis in human leukemia cells. Such sensitization is closely associated with the inhibitory effect of HA14-1 on TNF-alpha-mediated NF kappaB activation. HA14-1 suppressed NF-kappaB activation through inhibition of phosphorylation and degradation of IkappaBalpha. This inhibition was correlated with suppression of NF-kappaB-dependent gene products (c-myc, cyclin D1, cox-2, and IAP-1). Additionally, the present findings provide evidence of a critical role of ROS accumulation induced by HA14-1 in TNF-alpha-induced apoptosis. Moreover, HA14-1 also markedly sustained TNF-alpha-mediated JNK activation. A specific JNK inhibitor abolished the sensitization effect of HA14-1 on TNF-alpha induced apoptosis. Taken together, these results indicate that ROS and JNK represent important signals in HA14-1 sensitization in TNF-alpha-induced apoptosis. PMID- 20022692 TI - Removal of copper(II) ions by a biosorbent--Cinnamomum camphora leaves powder. AB - In the present study, Cinnamomum camphora leaves powder (CLP) was investigated as a biosorbent for the removal of copper ions from aqueous solutions. The biosorbents before and after adsorption were measured by EDS and FT-IR. Kinetic data and sorption equilibrium isotherms were carried out in batch process. The adsorption kinetic experiments revealed that there are three stages in the whole adsorption process. It was found that Cu(II) adsorption onto CLP for different initial Cu(II) concentrations all followed pseudo-second order kinetics and were mainly controlled by the film diffusion mechanism. Batch equilibrium results at different temperatures suggest that Cu(II) adsorption onto CLP can be described perfectly with Langmuir isotherm model compared to Freundlich and D-R isotherm models, and the characteristic parameters for each adsorption isotherm were also determined. Thermodynamic parameters calculated show that the adsorption process has been found to be endothermic in nature. The analysis for the values of the mean free energies of adsorption (E(a)), the Gibbs free energy (DeltaG(0)) and the effect of ionic strength all demonstrate that the whole adsorption process is mainly dominated by ion-exchange mechanism, accompanied by a certain amount of surface complexation which has been verified by variations in EDS and FT-IR spectra and pH value before and after adsorption. Regeneration studies show CLP possesses an excellent reusability. PMID- 20022693 TI - Quantitative risk assessment of CO2 transport by pipelines--a review of uncertainties and their impacts. AB - A systematic assessment, based on an extensive literature review, of the impact of gaps and uncertainties on the results of quantitative risk assessments (QRAs) for CO(2) pipelines is presented. Sources of uncertainties that have been assessed are: failure rates, pipeline pressure, temperature, section length, diameter, orifice size, type and direction of release, meteorological conditions, jet diameter, vapour mass fraction in the release and the dose-effect relationship for CO(2). A sensitivity analysis with these parameters is performed using release, dispersion and impact models. The results show that the knowledge gaps and uncertainties have a large effect on the accuracy of the assessed risks of CO(2) pipelines. In this study it is found that the individual risk contour can vary between 0 and 204 m from the pipeline depending on assumptions made. In existing studies this range is found to be between <1m and 7.2 km. Mitigating the relevant risks is part of current practice, making them controllable. It is concluded that QRA for CO(2) pipelines can be improved by validation of release and dispersion models for high-pressure CO(2) releases, definition and adoption of a universal dose-effect relationship and development of a good practice guide for QRAs for CO(2) pipelines. PMID- 20022691 TI - Cross-talk between miRNA and Notch signaling pathways in tumor development and progression. AB - Notch signaling pathways are known to regulate many cellular processes, including cell proliferation, apoptosis, migration, invasion, and angiogenesis, and is one of the most important signaling pathway during normal development. Recently, emerging evidences suggest that microRNAs (miRNAs) can function as key regulators of various biological and pathologic processes during tumor development and progression. Notch signaling has also been reported to be regulated through cross talk with many pathways and factors where miRNAs appears to play a major role. This article will provide a brief overview of the published evidences for the cross-talks between Notch and miRNAs. Further, we summarize how targeting miRNAs by natural agents could become a novel and safer approach for the prevention of tumor progression and treatment. PMID- 20022694 TI - Removal of nickel on Bofe bentonite calcined clay in porous bed. AB - Bentonite clays have been showing good adsorbing characteristics and are used as an alternative material in the removal of heavy metals. The purpose of this study is to evaluate the removal of nickel on Bofe bentonite calcined clay in porous bed. Firstly, a study was conducted to define the operation outflow, based on the minimum mass transfer zone (MTZ) obtained, useful (q(U)) and total adsorbed (q(T)) removal amounts and total nickel removal percentage (Rem (%)). Assays of nickel adsorption on clay were conducted according to a 2(2) factorial design with three central points to evaluate the effect of the particle diameter and initial adsorbate concentration on variables q(U), q(T) and Rem (%). Tests to obtain the adsorbent physical and chemical characteristics were performed on samples of Bofe clay in natura, calcined, and calcined submitted to nickel adsorption. This clay was characterized according to the following techniques: Energy Dispersive Spectroscopy (EDS), Thermogravimetry (TG), Differential Scanning Calorimetry (DSC), X-ray Diffraction (XRD), Fourier Transformed Infrared Spectroscopy (FTIR), Physisorption of N(2) (BET), Helium Picnometry and Scanning Electron Microscope (SEM) with metal mapping. PMID- 20022695 TI - Characteristics of organic, nitrogen and phosphorus species released from ultrasonic treatment of waste activated sludge. AB - Batch ultrasonic treatments (sonication) were performed on two waste activated sludge (WAS) samples, BNR-WAS from the biological nitrogen removal unit and BNPR WAS from the biological nitrogen and phosphorus removal unit of two Shanghai municipal WWTPs, to determine the effects of sonication time and intensity on the amount and distribution of the organic, N and P species released from the samples. The concentration profiles of COD, TOC fractions in different molecular weight (MW) ranges (<2 kDa, 2-100 kDa, and >100 kDa), TN, organic-N, NH(3)-N, TP and PO(4)-P were monitored during the treatment at three sonication intensity levels (0.167, 0.330 and 0.500 W/mL). Species releases increased with sonication time and/or intensity; the release rates were accelerated when the sonication intensity was above a critical level between 0.330 and 0.500 W/mL. After 1 h of treatment, 37.9%, 37.5% and 50.8% of the organic content (measured as COD) of BNR WAS were released, while the same for BNPR-WAS were 40.9%, 55.3% and 56.9%. It also resulted in the release of 40.9%, 38.7%, and 52.1% of total nitrogen from BNR-WAS, relative to 46.2%, 61.6%, and 70.4% of the same from BNPR-WAS; most released nitrogen were organic-N (65.0% and 84.9%), followed by NH(3)-N (34.7% and 14.9%) and trace amounts of nitrate and nitrite. More total phosphorus of a higher orthophosphate content was released from BNRP-WAS (>60% release after 1 h of sonication, 80% was PO(4)-P) than from BNR-WAS (<50% release, 40% was PO(4) P). The differences in the releases as well as the molecular weight distribution pattern of the soluble TOC species were due to the different structure and composition of the sludge samples. Sonication is a viable sludge treatment process when it is combined with a phosphorus recovery process to remove most of the released PO(4)-P so that the supernatant may be returned for further biological treatment. PMID- 20022696 TI - Habituation, sensitization, and emotional valence modulation of pain responses. AB - The Emotional Controls of Nociception (ECON) paradigm involves the presentation of emotionally-charged pictures during which painful stimuli are delivered. Across several ECON studies, unpleasant pictures enhanced pain and nociception, whereas pleasant pictures inhibited pain and nociception. However, at this time it is unknown whether emotional valence (unpleasant, neutral, pleasant) influences the habituation or sensitization of pain responses that occurs within a testing session. Indeed, ECON assumes that emotional valence modulation of pain is consistent throughout testing; otherwise the interpretation of valence modulation (unpleasant>neutral>pleasant) could be threatened. To address this issue, the present study (N=120) presented 108 pictures that varied in emotional valence. During and in between pictures, 52 suprathreshold electrocutaneous stimuli were delivered to evoke pain, the nociceptive flexion reflex [NFR], and pain-evoked skin conductance response [SCR]. Mixed effects ANOVAs verified that within-subject changes in pain responses were influenced by stimulus repetition (NFR and SCR habituated, pain ratings sensitized) and emotional valence (responses were highest during unpleasant pictures, intermediate during neutral pictures, and lowest during pleasant pictures). However, habituation/sensitization slopes were unaffected by emotional valence, thus indicating emotional valence modulation was consistently observed throughout the testing session. These results provide additional validation for the ECON paradigm and suggest that the circuit responsible for emotional modulation of pain and nociception is less susceptible to habituation or sensitization than the circuits responsible for responses to suprathreshold shocks. PMID- 20022697 TI - Distinctiveness of psychological obstacles to recovery in low back pain patients in primary care. AB - Many psychological factors have been suggested to be important obstacles to recovery from low back pain, yet most studies focus on a limited number of factors. We compared a more comprehensive range of 20 factors in predicting outcome in primary care. Consecutive patients consulting 8 general practices were eligible to take part in a prospective cohort study; 1591 provided data at baseline and 810 at 6 months. Clinical outcome was defined using the Roland and Morris Disability Questionnaire (RMDQ). The relative strength of the baseline psychological measures to predict outcome was investigated using adjusted multiple linear regression techniques. The sample was similar to other primary care cohorts (mean age 44 years, 59% women, mean baseline RMDQ 8.6). The 20 factors each accounted for between 0.04% and 33.3% of the variance in baseline RMDQ score. A multivariate model including all 11 scales that were associated with outcome in the univariate analysis accounted for 47.7% of the variance in 6 months RMDQ score; rising to 55.8% following adjustment. Four scales remained significantly associated with outcome in the multivariate model explaining 56.6% of the variance: perceptions of personal control, acute/chronic timeline, illness identify and pain self-efficacy. When all independent factors were included, depression, catastrophising and fear avoidance were no longer significant. Thus, a small number of psychological factors are strongly predictive of outcome in primary care low back pain patients. There is clear redundancy in the measurement of psychological factors. These findings should help to focus targeted interventions for back pain in the future. PMID- 20022698 TI - NGF induces non-inflammatory localized and lasting mechanical and thermal hypersensitivity in human skin. AB - Nerve growth factor (NGF) modulates sensitivity and sprouting of nociceptors. We explored the spatial and temporal sensitization induced by NGF injection (1 microg) in human skin. Hyperalgesia was investigated in 16 volunteers (36+/-9 years) at day 1, 3, 7, 21, and 49. Areas of mechanical (brush, pin-prick) and heat (43 degrees C) sensitization were mapped and thermal (heat and cold) pain thresholds, mechanical (impact stimulation) and electrically evoked pain, and axon reflex flare were assessed. No spontaneous pain or local inflammation was recorded upon NGF injection and during 49 days. Sensitization to heat was maximum at day 3 and lasted 21 days. Hyperalgesia to cold was recorded at day 7 and 21. Hypersensitivity to mechanical impact stimuli developed delayed, reached maximum at day 21, and persisted throughout 49 days. Fifty percent of all volunteers reported a static allodynia to tonic pressure until day 21. Electrical stimulation at 7.5 mA was more painful at the NGF site at day 21, which correlated significantly to maximum impact pain. Axon reflex flare was unaffected by NGF. Sensitization was limited to the NGF injection site, no touch- or pin prick evoked secondary hyperalgesia was observed. Spatially restricted hyperalgesia indicates a peripheral rather than central mechanism. The temporal profile of lasting nociceptor sensitization suggests an altered peripheral axonal expression of sensory proteins specifically leading to mechanical and thermal sensitization. Intradermal NGF administration provokes a pattern of sensitization that can be used as experimental model for neuropathic pain. PMID- 20022700 TI - Milk and no sugar. PMID- 20022701 TI - An unusual exit point from an electrocution injury. PMID- 20022699 TI - Aberrant sensory responses are dependent on lesion severity after spinal cord contusion injury in mice. AB - Following spinal cord injury (SCI), individuals lose normal sensation and often develop debilitating neuropathic pain. Basic research has helped to elucidate many of the underlying mechanisms, but unanswered questions remain concerning how sensation changes after SCI and potential negative consequences of regenerative therapies. Mouse models provide an opportunity to explore these questions using genetic markers and manipulations. However, despite the increasing use of mice in pain and sensory research, the responses to sensory stimuli after SCI are poorly characterized in this species. This study evaluated behavioral responses to mechanical and nociceptive stimuli applied to the hindlimbs and the dorsal trunk in C57BL/6 mice after mid-thoracic SCI. Adult mice were subjected to laminectomy, contusion injuries of different severities, or complete transections to test the hypothesis that the patterns of sensory pathology depend on the extent of tissue damage at the injury site. In the hind paws, hyper-responsiveness to a heat stimulus developed independent of injury severity, while mechanical sensitivity decreased, except after the most severe contusion injuries sparing less than 2% of the white matter at the injury site, when enhanced sensitivity was observed. On the trunk, mechanical and pin prick testing revealed diminished sensitivity at and below the injury level, while responses above the level of the injury were unchanged. The contrast in injury severity threshold for thermal and mechanical hypersensitivity in the hind paws suggests that these responses have different underlying mechanisms. These results establish essential baseline information for murine studies of pain and changes in sensation after SCI. PMID- 20022702 TI - Tissue engineering of skin. AB - The engineering of skin substitutes and their application on human patients has become a reality. However, cell biologists, biochemists, technical engineers, and surgeons are still struggling with the generation of complex skin substitutes that can readily be transplanted in large quantities, possibly in only one surgical intervention and without significant scarring. Constructing a dermo epidermal substitute that rapidly vascularizes, optimally supports a stratifying epidermal graft on a biodegradable matrix, and that can be conveniently handled by the surgeon, is now the ambitious goal. After all, this goal has to be reached coping with strict safety requirements and the harsh rules of the economic market. PMID- 20022704 TI - Laptop computer-induced thigh burn. PMID- 20022705 TI - Comments on "Platelet-rich plasma in burns". PMID- 20022706 TI - Quality assessment of clinical practice guidelines for adaptation in burn injury. AB - OBJECTIVES: A number of clinical practice guidelines (CPGs) are available for managing burn injury patients but clinical practice is highly variable. We report the first steps to trans-contextual adaptation of international burn CPGs to local settings. METHODS: Key clinical topics and questions to be covered in the final guideline were defined and prioritized. Systematic search between 1990 and 2008 retrieved 546 citations, of which 24 were CPGs on the general and intensive care of burn patients. Assessment of the clinical content of CPGs was carried out. Methodological quality of CPGs was evaluated using the AGREE instrument. RESULTS: Of the 24 CPGs evaluated, 10 (42%) were evidence-based. All major burn topics were covered by at least one CPG, but no single CPG addressed all areas important in terms of outcomes. According to the AGREE criteria, 2 CPGs (8%) were strongly recommended, 14 with provisos or alterations (58%) and the rest were not recommended for adaptation. CONCLUSIONS: Although existing CPGs for the management of burn may accurately reflect agreed clinical practice, most performed poorly when evaluated for methodological quality. Future CPG efforts addressing these methodological shortcomings would add substantially to the improved management of burned patients. PMID- 20022707 TI - Title to a text is author's prerogative. PMID- 20022708 TI - The treatment of deep dermal hand burns: How do we achieve better results? Should we use allogeneic keratinocytes or skin grafts? AB - The treatment of deep dermal burns has a broad spectrum and has been subject to discussion over the past years. The treatment of hand burns is challenging due to the high requirements to aesthetic and functional outcome. 27 patients, 7 women and 20 men with deep dermal hand burns with a mean age of 41.3+/-16.5 and a mean TBSA of 15%+/-19.6% were treated either with allogeneic cryopreserved keratinocytes or with split skin grafts. Long-term follow-up revealed no statistical significant differences between the two groups concerning Vancouver Scar Scale as well as hand function judged by the DASH score; however there was a tendency to higher VSS scores and impaired aesthetic results in the keratinocyte group. Allogeneic keratinocytes are a suitable armentarium for the treatment of deep dermal hand burns; and, if used correctly, they can produce a timely healing comparable to split-thickness skin grafts. Limited availability, high costs as well as the need for special skills are key factors, which render application of this technique outside specialist burn centres virtually impossible. In our opinion, the cultivation and use of keratinocytes should be reserved to these centres in order to facilitate a sensible application for a full range of indications. We recommend usage of allogeneic keratinocytes for deep dermal hand burns only in severely burned patients with a lack of donor sites. Patients with unrestricted availability of donor sites seem to profit from the application of split-thickness skin grafts according to our results. PMID- 20022710 TI - Could the urolithiasis be treated by anti-nanobacterial therapy? PMID- 20022709 TI - Forkhead box M1 transcription factor: a novel target for cancer therapy. AB - FoxM1 signaling has been reported to be associated with carcinogenesis. Therefore, the FoxM1 may represent a novel therapeutic target, and thus the development of agents that will target FoxM1 is likely to have significant therapeutic impact on human cancer. This review describes the mechanisms of signal transduction associated with FoxM1 and provides emerging evidence in support of its role in the carcinogenesis. Further, we summarize data on several FoxM1 inhibitors especially "chemopreventive agents" and these agents could be useful for targeted inactivation of FoxM1, which indeed could become a novel approach for the prevention and/or treatment of human cancer. PMID- 20022711 TI - Follow-up motion laboratory analysis for patients with spastic hemiplegia due to cerebral palsy: analysis of the flexor carpi ulnaris firing pattern before and after tendon transfer surgery. AB - PURPOSE: To compare the preoperative and postoperative pattern of firing of the flexor carpi ulnaris (FCU) in a grasp and release functional activity for children treated with an FCU to extensor carpi radialis brevis tendon transfer for wrist flexion deformity associated with spastic hemiplegia from cerebral palsy. METHODS: Seven children, evaluated by a preoperative EMG video analysis and treated with an FCU to extensor carpi radialis brevis transfer, had a follow up postoperative EMG/video motion laboratory analysis at an average follow-up of 3.5 years (range, 1.0-6.8 years). Each preoperative and postoperative EMG/video was reviewed for the task of lifting heavy cans, as described by Jebson et al. The EMG activity of the FCU was described as active or relaxed during grasp and during release. RESULTS: Preoperatively, the most common pattern was to activate the FCU during grasp and to relax the FCU during release (4 patients). Postoperatively, 6 patients activated the FCU during grasp and relaxed the FCU during release; 1 patient activated the FCU during both grasp and release. CONCLUSIONS: Of the 7 patients studied, the FCU changed phase from preoperative to postoperative in only 1. This study concludes that most commonly the FCU does not predictably change phase when transferred from a position of wrist flexion to wrist extension. TYPE OF STUDY/LEVEL OF EVIDENCE: Therapeutic IV. PMID- 20022712 TI - Dynamics of intracarpal tunnel pressure in patients with carpal tunnel syndrome. AB - PURPOSE: To measure pressure within the carpal tunnel (intracarpal tunnel pressures) in patients with carpal tunnel syndrome and determine the effect of quantified active hand use on both the magnitude and location of peak pressures, before and after division of the transverse carpal ligament. METHODS: We measured intracarpal tunnel pressures intraoperatively in 12 patients with carpal tunnel syndrome at 5 standardized locations based on the distance between each patient's proximal pisiform and hook of hamate (HH) before endoscopic division of the ligament, using a semiconductor gauge pressure sensor inserted from proximal to distal into the tunnel under fluoroscopic control. At each location, pressure was recorded with fingers extended, fingers flexed, and 50% maximum grip using a grip dynamometer. Additional hand use activities, including maximum key and pulp pinch using a pinch dynamometer, 25% maximum grip, and maximum grip, were performed by a subset of these patients. After ligament division, we measured pressures during the same hand activities at a single location, HH. We analyzed the effect of hand activity, measurement location, and ligament division using repeated measures analysis of variance. RESULTS: Compared with fingers extended (mean pressure, 56 mm Hg), all pinch and grip activities caused significant increases in pressure at HH, with a mean peak pressure of 1151 mm Hg during maximum grip. After endoscopic release, pressures decreased significantly at HH for all hand activities. CONCLUSIONS: In patients with carpal tunnel syndrome, intracarpal tunnel pressures during active hand use are substantially greater than previously reported. Peak pressures occur at the HH, where the tunnel is most constricted and the median nerve is most compressed in carpal tunnel syndrome. TYPE OF STUDY/LEVEL OF EVIDENCE: Therapeutic IV. PMID- 20022713 TI - Medium chain fatty acid feed supplementation reduces the probability of Campylobacter jejuni colonization in broilers. AB - Campylobacteriosis in humans is associated with handling and consumption of contaminated broiler meat. Reduction of the number of Campylobacter-colonized broiler flocks could potentially be realized by decreasing their susceptibility for colonization. The aim of this study was to determine the effect of feed supplementation with a mixture of medium chain fatty acids (C(8)-C(12)) on susceptibility of broilers for Campylobacter colonization, feed conversion and body weight gain. Two experiments were carried out with individually housed commercial broilers. The birds were fed with medium chain fatty acids supplemented feed (n=227), or received feed without supplement (n=87). The birds were inoculated with a dose of Campylobacter jejuni varying between log(10)1.19 5.47 CFU. During 14 days after inoculation, cecal or fecal samples were collected, in which the presence of C. jejuni was determined by bacterial culture. Beta-binomial dose-response modeling of the colonization status at 14 days post-inoculation was performed to estimate the C. jejuni dose necessary to colonize 50% of inoculated broilers, which was estimated to be 200 times higher in broilers fed with supplemented feed (log(10)4.8 CFU) than in control broilers (log(10)2.5 CFU). Feed conversion was not affected by feed supplementation, while body weight gain was 49 g higher in broilers fed with supplemented feed. These findings indicate that susceptibility of broilers for Campylobacter colonization is decreased by supplementation with medium chain fatty acids, and that feed supplemented with this mixture may be a promising tool for the reduction of Campylobacter colonization in commercial broiler flocks. PMID- 20022714 TI - Isolation and genetic analysis revealed no predominant new strains of avian infectious bronchitis virus circulating in South China during 2004-2008. AB - Twenty-seven strains of avian infectious bronchitis virus (IBV) were isolated from dead or diseased chickens at different chicken farms in South China during 2004-2008, of which the S1 gene was sequenced. Phylogenetic analysis of the S1 gene sequences of the isolated 27 strains together with 29 strains published in Genbank revealed that all IBV strains except for one isolated and one published were clustered into six distinct genotypes I-VI. 26 isolated strains belong to genotypes I, II, and III, forming a big phylogenetic branch without new predominant strains, whereas all five vaccine strains belong to genotype V that is evolutionarily distant from genotypes I, II, and III. The study of the protease cleavage motif within the S1 protein found 12 different cleavage motifs, of which 3 motifs are shared by both isolated and published strains, 2 motifs unique to isolated strains, and 7 motifs unique to published strains, further bolstering the notion of no new predominant strains. Alignment analysis of the S1 amino acid sequences indicated that the amino acid substitutions, insertions, and deletions are polymorphic and diverse, showing no sign of predominant genetic changes among the isolated strains. Taken together, there was no predominant new strain circulating in South China during 2004-2008. Nonetheless, circulating IBV strains have been continuously evolving with genetic compositions distant from vaccine strains; this explains why there have been constant but infrequent outbreaks in commercial flocks in South China during 2004-2008. Furthermore, in order to safe guard against the sudden emergence of new predominant strains, continuing surveillance of IBV strains circulating in the field is of extreme importance. PMID- 20022715 TI - Dietary aluminosilicate supplement enhances immune activity in mice and reinforces clearance of porcine circovirus type 2 in experimentally infected pigs. AB - Aluminosilicate is the major component of clay minerals such as zeolite, bentonite and clinoptilolite. The minerals possess a number of beneficial activities, especially in regulating the immune system. The aims of the present study were to evaluate immune enhancing effects of dietary aluminosilicate supplement (DAS) in mice, and to demonstrate clearance effects of DAS against porcine circovirus type 2 (PCV2) in experimentally infected pigs as an initial step towards the development of an antibiotic substitute for use in pigs. Relative messenger RNA expression levels of interferon-gamma, interleukin-4 and tumor necrosis factor-alpha, phagocytic activities of polymorphonuclear leucocytes, serum antibody production level and spleen B cell ratio were significantly increased in the DAS groups of mice compared with the control group (each feeding group had three replications with 5 mice each). The results indicated that general immune activity including cellular and humoral immunity could be enhanced by DAS in mice. In experimentally PCV2-infected pigs, the load of viral genome in nasal swab, serum and lung of the DAS group of pigs was significantly decreased compared with the control group at 28 days post-infection (each group three pigs). Corresponding histopathological analyses demonstrated that pigs in the DAS group displayed mild and less severe abnormal changes compared with the control group, indicating that DAS reinforces clearance of PCV2 in experimentally infected pigs. This may relate to general immune enhancing effects of DAS in mice. Therefore DAS will help the health of animal, especially in swine. PMID- 20022716 TI - An update on safety studies on the attenuated "RIEMSER Schweinepestoralvakzine" for vaccination of wild boar against classical swine fever. AB - The RIEMSER Schweinepestoralvakzine is an attenuated vaccine for oral vaccination of wild boar against classical swine fever (CSF). The safety of this licensed bait vaccine which is based on the CSF virus (CSFV) strain "C" was investigated in eight animal species, e.g. weaner pigs (n=111), wild boar (n=11), ruminants (cattle, goats and sheep, n=11), foxes (n=5), rabbits (n=12), and mice (n=10). Animals were vaccinated either with a single vaccine dose containing at least 10(4.5) TCID(50), or with overdoses, i.e. the 10-fold dose, or they were subjected to repeated application schemes. During the entire observation period none of the animals which were given the vaccine virus showed clinical signs, with the exception of rabbits. These reacted to the vaccination with fever. Orally vaccinated pigs did not transmit vaccine virus to susceptible contact animals (sentinels). In none of the species examined neither vaccine virus nor viral RNA could be detected in blood after vaccination. In one wild boar viral RNA could be established in the tonsil 21 days post-vaccination (dpv); all other organ samples tested virologically negative. Up to 77.5% of the pigs and wild boar developed virus neutralising antibodies (VNA) already 14 dpv. The mean VNA titres observed in the vaccination groups seemed to depend rather on individual factors than on the administered virus dose (virus titre per dose) or the vaccination scheme. These results are comparable with findings obtained during oral vaccination campaigns in wild boar and after parenteral vaccination with this C-strain virus. From the results presented here it can be concluded that RIEMSER Schweinepestoralvakzine is safe for target and non-target species. PMID- 20022717 TI - Massage therapy improves neurodevelopment outcome at two years corrected age for very low birth weight infants. AB - BACKGROUND: Long term effects of massage therapy in very preterm newborns infants are still to be described. Few studies evaluated neurodevelopment just at six months, and included late preterm infants. OBJECTIVE: To study the effect of massage therapy on neurodevelopment of very low birth weight infants at two years corrected age. STUDY DESIGN: Newborns with birth weight between >or= 750 and 70 mm, puncture points 10 mm from the midtragus led to insertion of the arthroscope inside the upper joint compartment. For measurements < or = 70 mm, puncture points 7 mm from the midtragus led to insertion of the arthroscope inside the upper joint compartment. This suggests that for canthal-tragus distances of > 70 mm, the arthroscope should be inserted 10mm from the midtragus and for distances < or = 70 mm it should be inserted at 7 mm for the greatest likelihood of entering the upper joint compartment of the TMJ. PMID- 20022728 TI - ICOS expression by effector T cells influences the ability of regulatory T cells to inhibit anti-chromatin B cell responses in recipient mice. AB - T regulatory cells are critical for the prevention of autoimmunity. Specifically, Treg cells can control anti-chromatin antibody production in vivo, and this correlates with decreased ICOS expression on CD4(+) T helper cells. Here we test the significance of high ICOS expression by T effector cells, firstly in terms of the anti-chromatin B cell response, and secondly on the ability of Treg cells to suppress T cell help. We bred CD4(+) T cell receptor transgenic mice with mice that carry the Roquin(san/san) mutation. The Roquin gene functions to limit ICOS mRNA such that CD4 T cells from mutant mice express elevated ICOS. Using an in vivo model, TS1.Roquin(san/san) Th cells were compared with wild-type TS1 Th cells with regard to their ability to help anti-chromatin B cells in the presence or absence of Treg cells. Both TS1 and TS1.Roquin(san/san) Th cells induced anti chromatin IgM(a) antibodies, but the TS1.Roquin(san/san) Th cells resulted in the recovery of more class-switched and germinal center B cells. Neither source of Th cells were capable of inducing long-lived autoantibodies. Treg cells completely suppressed anti-chromatin IgM(a) antibody production and reduced anti-chromatin B cell recovery induced by TS1 Th cells. Importantly, this suppression was less effective when TS1.Roquin(san/san) Th cells were used. Thus, high ICOS levels on effector T cells results in autoimmunity by augmenting the autoreactive B cell response and by dampening the effect of Treg cell suppression. PMID- 20022730 TI - Thinking about semantic concepts in schizophrenia: the more familiar the less deviation. PMID- 20022731 TI - GABA concentration in schizophrenia patients and the effects of antipsychotic medication: a proton magnetic resonance spectroscopy study. AB - Gamma-amino butyric acid (GABA) is thought to play a role in the pathophysiology of schizophrenia. High magnetic field proton magnetic resonance spectroscopy ((1)H-MRS) provides a reliable measurement of GABA in specific regions of the brain. This study measured GABA concentration in the anterior cingulate cortex (ACC) and in the left basal ganglia (ltBG) in 38 patients with chronic schizophrenia and 29 healthy control subjects. There was no significant difference in GABA concentration between the schizophrenia patients and the healthy controls in either the ACC (1.36+/-0.45 mmol/l in schizophrenia patients and 1.52+/-0.54 mmol/l in control subjects) or the ltBG (1.13+/-0.26 mmol/l in schizophrenia patients and 1.18+/-0.20 mmol/l in control subjects). Among the right handed schizophrenia patients, the GABA concentration in the ltBG was significantly higher in patients taking typical antipsychotics (1.25+/-0.24 mmol/l) than in those taking atypical antipsychotics (1.03+/-0.24 mmol/l, p=0.026). In the ACC, the GABA concentration was negatively correlated with the dose of the antipsychotics (rs=-0.347, p=0.035). In the ltBG, the GABA concentration was positively correlated with the dose of the anticholinergics (rs=0.403, p=0.015). To the best of our knowledge, this is the first study to have directly measured GABA concentrations in schizophrenia patients using (1)H MRS. Our results suggest that there are no differences in GABA concentrations in the ACC or the ltBG of schizophrenia patients compared to healthy controls. Antipsychotic medication may cause changes in GABA concentration, and atypical and typical antipsychotics may have differing effects. It is possible that medication effects conceal inherent differences in GABA concentrations between schizophrenia patients and healthy controls. PMID- 20022732 TI - Grafting of chitosan as a biopolymer onto wool fabric using anhydride bridge and its antibacterial property. AB - Weak binding of chitosan on the wool constitutes the main problem in its application. In this paper, the surface modification of wool fabric using anhydrides to graft the chitosan was studied. Weight gain, antibacterial and antifelting properties of the chitosan grafted-acylated wool fabric were investigated. Wool fabrics were acylated with two anhydrides, succinic anhydride (SA) and phthalic anhydride (PA), using different solvents (dimethylsulfoxide (DMSO) and N,N-dimethyl formamide (DMF)). The effects of anhydrides, solvents, anhydride concentration, liquor ratio (L:R) and reaction time on acylation of wool were investigated. Chitosan was grafted to the acylated wool and the effects of pH, chitosan concentration, and reaction time on chitosan grafting of acylated wool were evaluated. Fourier transform infra-red (FTIR), scanning electron microscope (SEM), differential scanning colorimetry (DSC) and weight gain analyses provided evidence that chitosan was grafted on to the acylated wool through the formation of new covalent bonds. The grafted samples have antibacterial potential due to existence of the antibacterial property of chitosan. In addition, the chitosan grafted-acylated wool samples have antifelting property. The findings of this research support the potential production of new environmentally friendly textile fabrics. PMID- 20022733 TI - Mildly abnormal general movement quality in infants is associated with higher Mead acid and lower arachidonic acid and shows a U-shaped relation with the DHA/AA ratio. AB - We showed that docosahexaenoic acid (DHA) supplementation during pregnancy and lactation was associated with more mildly abnormal (MA) general movements (GMs) in the infants. Since this finding was unexpected and inter-individual DHA intakes are highly variable, we explored the relationship between GM quality and erythrocyte DHA, arachidonic acid (AA), DHA/AA and Mead acid in 57 infants of this trial. MA GMs were inversely related to AA, associated with Mead acid, and associated with DHA/AA in a U-shaped manner. These relationships may indicate dependence of newborn AA status on synthesis from linoleic acid. This becomes restricted during the intrauterine period by abundant de novo synthesis of oleic and Mead acids from glucose, consistent with reduced insulin sensitivity during the third trimester. The descending part of the U-shaped relation between MA GMs and DHA/AA probably indicates DHA shortage next to AA shortage. The ascending part may reflect a different developmental trajectory that is not necessarily unfavorable. PMID- 20022734 TI - Hypoxia-induced autophagy: cell death or cell survival? AB - Hypoxia (approximately 3-0.1% oxygen) is capable of rapidly inducing, via the hypoxia-inducible factor (HIF-1), a cell survival response engaging autophagy. This process is mediated by the atypical BH3-only proteins the Bcl-2/E1B 19kDa interacting protein 3 (BNIP3/BNIP3L (NIX)) that are induced by HIF-1. These mitochondrial associated BNIP proteins also mediate mitophagy, a metabolic adaptation for survival that is able to control reactive oxygen species (ROS) production and DNA damage. In contrast, severe hypoxic conditions or anoxia (<0.1% oxygen), where the latter is often confused with physiological hypoxia, are capable of inducing a HIF-independent autophagic response, generated via an extreme nutritional stress response implicating the AMPK-mTOR and unfolded protein response (UPR) pathways. The autophagic cell death that is often observed in these extreme stress conditions should be seen as the outcome of failed adaptation. PMID- 20022736 TI - New look inside the spindle: microtubule-dependent microtubule generation within the spindle. AB - The structure, dynamics, and mechanics of mitotic and meiotic spindles have been progressively elucidated through the advancements in microscopic technology, identification of the genes involved, and construction of theoretical frameworks. Here, we review recent works that have utilized quantitative image analysis to advance our understanding of the complex spindle structure of animal cells. In particular, we discuss how microtubules (MTs) are nucleated and distributed inside the spindle. Accumulating evidence supports the presence of MT-dependent MT generation within the spindle. This mechanism would produce dense arrays of intraspindle MTs with various lengths, which may contribute to efficient spindle assembly and stabilize the metaphase spindle. RNA interference (RNAi) screens with quantitative image analysis led to the identification of the augmin complex that plays a key role in this MT generation process. PMID- 20022735 TI - Mechanochemical crosstalk during endocytic vesicle formation. AB - Membrane curvature has emerged as a key regulatory factor in endocytic vesicle formation. From a theoretical perspective, we summarize recent progress in understanding how membrane curvature and biochemical pathways are coupled and orchestrated during the coherent process of endocytic vesicle formation. We mainly focus on clathrin-mediated and actin-mediated endocytosis in yeast and in mammalian cells. We further speculate on how mechanochemical feedback could modulate other membrane-remodeling processes. PMID- 20022737 TI - Recycling of rubble from building demolition for low-shrinkage concretes. AB - In this project concrete mixtures were prepared that were characterized by low ductility due to desiccation by using debris from building demolition, which after a suitable treatment was used as aggregate for partial replacement of natural aggregates. The recycled aggregate used came from a recycling plant, in which rubble from building demolition was selected, crushed, cleaned, sieved, and graded. Such aggregates are known to be more porous as indicated by the Saturated Surface Dry (SSD) moisture content. The recycled concrete used as aggregates were added to the concrete mixture in order to study their influence on the fresh and hardened concrete properties. They were added either after water pre-soaking or in dry condition, in order to evaluate the influence of moisture in aggregates on the performance of concrete containing recycled aggregate. In particular, the effect of internal curing, due to the use of such aggregates, was studied. Concrete behavior due to desiccation under dehydration was studied by means of both drying shrinkage test and German angle test, through which shrinkage under the restrained condition of early age concrete can be evaluated. PMID- 20022738 TI - Attenuation of hydrogen sulfide at construction and demolition debris landfills using alternative cover materials. AB - The attenuation of H(2)S emissions by various landfill cover materials was evaluated using both laboratory and field experiments. The results demonstrated that cover materials consisting of selected waste products (compost and yard trash) and soils amended with quicklime and calcium carbonate effectively attenuated H(2)S emissions and detectable H(2)S emissions were only encountered in a testing plot using a sandy soil cover (average emission rate was 4.67x10( 6)mgm(-2)s(-1)). H(2)S concentration profiles in the cover materials indicated that H(2)S was removed as it migrated through the cover materials. At the same depth in the testing area, the H(2)S concentration in the sandy soil field plot was always higher than that of other testing plots because the sand (a) demonstrated less ability to remove H(2)S and (b) exhibited a higher H(2)S concentration at the base of the cover. Laboratory experiments confirmed these observations, with a combination of physical adsorption, chemical reactions, and biological oxidation, accounting for the enhanced removal. In addition to removal, the results suggest that some of the cover materials reduced H(2)S generation by creating less favorable conditions for sulfate-reducing bacteria (e.g., high pH and temperature). PMID- 20022739 TI - An investigation on pollutant emissions from co-firing of RDF and coal. AB - Pollutant emissions from co-firing of refuse derived fuel (RDF) and coal were investigated in a vortexing fluidized bed combustor (VFBC). RDF-5 was made of common municipal solid waste (MSW). CaCO(3) was injected in the combustor to absorb HCl at 850 degrees C. The results show that NO(x) and HCl emissions increase with RDF-5 co-firing ratio. The NO(x) concentration in flue gas at the bottom of the combustor is higher than that at the top. However, the trend of HCl released is reverse compared with NO(x) emissions. It was found that the HCl concentration decreases with increasing the molar ratio of Ca/Cl. However, the effect of CaCO(3) addition on HCl retention is not significant when the molar ratio of Ca/Cl is higher than 5. The chlorine content in fly ash increases obviously with the molar ratio of Ca/Cl. PCDD/Fs emissions decrease slightly with an addition of CaCO(3). In this study incomplete combustion is regarded as the main cause for polychlorinated dibenzo-p-dioxins and dibenzofurans (PCDD/Fs) formation. PMID- 20022740 TI - Prevention of lead leaching from fly ashes by mechanochemical treatment. AB - Fly ashes from a municipal solid waste incinerator were subjected to mechanochemical (MC) treatment in a planetary ball mill, and the treated fly ashes were cemented with a binder. The leachability of lead (Pb) from the treated fly ashes and from the cement product were investigated, and the speciation of lead in the treated and untreated ashes was determined by X-ray absorption fine structure (XAFS) analysis. MC treatment of the fly ashes and subsequent cementation prevented Pb leaching by 99.9%, whereas MC treatment alone prevented Pb leaching by 92.8%. Analysis of the X-ray absorption near-edge spectrum of the untreated fly ashes revealed that the predominant Pb species in the ashes was PbCl(2). In contrast, the counterpart in the treated fly ashes was Pb(3)O(4) insoluble in water. The formation of a species of Pb with a lower solubility in water than that of PbCl(2) was confirmed by MC treatment of PbCl(2)-spiked fly ashes for 48h, indicating the reduction of PbCl(2) in the spiked fly ashes to Pb via Pb(3)O(4) during MC treatment. Our results indicate that such reduction to an insoluble species prevented Pb from leaching and that MC treatment followed by cementation is a feasible method for the recycling of fly ashes. PMID- 20022741 TI - Simultaneous determination of five-type hepatitis virus antigens in 5 min using an integrated automatic electrochemical immunosensor array. AB - An integrated automatic electrochemical immunosensor array has been designed for the simultaneous detection of 5-type hepatitis virus antigens (i.e. hepatitis A, hepatitis B, hepatitis C, hepatitis D, and hepatitis E). Initially, 5-type hepatitis virus antibodies were immobilized onto a self-made electrochemical sensor array using nanogold particles and protein A as matrices, and the immunosensor array was then used to capture their corresponding antigens from sample solution with a 1-step capture format. The detection is based on the potential change before and after the antigen-antibody reaction by using a 2 electrode system. The developed immunosensor array with allowed simultaneous determination of 5-type hepatitis virus antigens in 5 min. The detection limit of the sensor array was 0.8). Cronbach's alpha was generally greater than 0.6, indicating that the reliability was high. Of the patients that responded, 27.3% preferred the electronic format, 25.7% preferred the paper format and 47% had no preference. The average time taken to complete a questionnaire was about 9 minutes for each format. The different questionnaire formats measured toxicity effects consistently and were reliable for both gynaecological cancer and prostate cancer patients. The survey indicated that patients found the questionnaires clear, easy to understand and straightforward to complete. Electronic data capture of patient-reported toxicity for CTCAE is feasible and acceptable. PMID- 20022743 TI - Improved vitamin B(12) production by step-wise reduction of oxygen uptake rate under dissolved oxygen limiting level during fermentation process. AB - Effects of different oxygen transfer rates (OTR) on the cell growth and vitamin B(12) biosynthesis of Pseudomonas denitrificans were first investigated under dissolved oxygen limiting conditions. The results demonstrated that high OTR accelerated cell growth and initial vitamin B(12) biosynthesis rate, while lower OTR was critical for higher productivity in the late fermentation process. The oxygen uptake rates (OUR) corresponded well with OTR. Based on the metabolic intermediate analysis, a step-wise OUR control strategy was proposed. The strategy was successfully implemented in scale-up to an industrial fermenter (120,000 l). A stable maximum vitamin B(12) production of 208 + or - 2.5 mg/l was achieved, which was increased by 17.3% compared with the control. Furthermore, the glucose consumption coefficient to vitamin B(12) was 34.4% lower than that of the control. An efficient and economical fermentation process based on OUR criterion was established for industrial vitamin B(12) fermentation by P. denitrificans. PMID- 20022744 TI - Supercritical carbon dioxide extraction of seed oil from yellow horn (Xanthoceras sorbifolia Bunge.) and its anti-oxidant activity. AB - Supercritical fluid carbon dioxide (SF-CO(2)) extraction (SFE) of seed oil from yellow horn and its anti-oxidant activity were investigated. The effects of CO(2) flow rate and particle size were firstly optimized, and a central composite design (CCD) combined with response surface methodology was used to study the effects of extraction pressure, temperature and time on the extraction yields. A maximal extraction yield of 61.28% was achieved under optimal conditions of extraction pressure 30 MPa at 45.68 degrees C, 2.08 h and CO(2) flow rate 12 kg/h with 0.5mm particle size. By analyzing the chemical composition of the seed oil, we found that the content of unsaturated fatty acids was approximately 90%. Furthermore, the anti-oxidant activity of seed oil was assessed by a 2,2-diphenyl 1-picrylhydrazyl (DPPH) radical-scavenging assay and a beta-carotene bleaching test. Yellow horn seed oil possessed notable concentration-dependent anti-oxidant activity with IC(50) values of 0.151 and 0.195 g/mL, respectively. PMID- 20022745 TI - Methanogenic activities in anaerobic membrane bioreactors (AnMBR) treating synthetic municipal wastewater. AB - Two laboratory-scale anaerobic membrane bioreactors, AnMBR 1 and AnMBR 2, were run in parallel at 25 and 15 degrees C, respectively. Total chemical oxygen demand (COD) removal efficiency was more than 95% and 85% for AnMBR 1 and 2, respectively. The COD removal of AnMBR 1 was mostly carried out biologically. However, the physical removal on the membrane surface compensated for the decreased biological removal rate in AnMBR 2. The membrane in AnMBR systems is likely not only to retain all biomass in the reactor, but also complement decreased biological removal efficiency at low temperature by rejecting soluble organics. Specific methanogenic activity (SMA) test was used to investigate the methanogenic activity profiles of suspended and attached sludge in AnMBRs treating synthetic municipal wastewater at 25 and 15 degrees C. The methanogenic activity was 51.8 ml CH(4)/g VSSd on day 1 and eventually increased 27% and reached 65.7 ml CH(4)/g VSSd on day 75 for AnMBR 1. However, the methanogenic activity of AnMBR 2 sludge was lower than that of AnMBR 1. The microbial activity of suspended sludge continuously increased, while that of attached sludge gradually decreased in this study. The methanogenic activity of attached sludge was far lower than that of suspended sludge. The role of attached sludge on the membrane in AnMBRs as a biofilm for biological organic removal was minimal compared to suspended sludge. PMID- 20022746 TI - Bioethanol from lignocelluloses: Status and perspectives in Brazil. AB - The National Alcohol Program--ProAlcool, created by the government of Brazil in 1975 resulted less dependency on fossil fuels. The addition of 25% ethanol to gasoline reduced the import of 550 million barrels oil and also reduced the emission CO(2) by 110 million tons. Today, 44% of the Brazilian energy matrix is renewable and 13.5% is derived from sugarcane. Brazil has a land area of 851 million hectares, of which 54% are preserved, including the Amazon forest (350 million hectares). From the land available for agriculture (340 million hectares), only 0.9% is occupied by sugarcane as energy crop, showing a great expansion potential. Studies have shown that in the coming years, ethanol yield per hectare of sugarcane, which presently is 6000 L/ha, could reach 10,000 L/ha, if 50% of the produced bagasse would be converted to ethanol. This article describes the efforts of different Brazilian institutions and research groups on second generation bioethanol production, especially from sugarcane bagasse. PMID- 20022747 TI - Development of substituted 6-[4-fluoro-3-(piperazin-1-ylcarbonyl)benzyl]-4,5 dimethylpyridazin-3(2H)-ones as potent poly(ADP-ribose) polymerase-1 (PARP-1) inhibitors active in BRCA deficient cells. AB - We describe an extensive SAR study in the 6-[4-fluoro-3-(substituted)benzyl]-4,5 dimethylpyridazin-3(2H)-one series which led to the identification of potent PARP 1 inhibitors, capable of inhibiting the proliferation of BRCA-1 deficient cancer cells in the low nanomolar range, and displaying >100-fold selectivity over the BRCA wild type counterparts. The series of compounds was devoid of hERG channel activity, and CYP inhibition and induction liabilities. Several analogs were stable in rat and human liver microsomes and displayed moderate rat clearance, with urinary excretion of parent as the major route of elimination. PMID- 20022748 TI - Molecular modeling study of 4-phenylpiperazine and 4-phenyl-1,2,3,6 tetrahydropyridine derivatives: a new step towards the design of high-affinity 5 HT1A ligands. AB - The main feature of many drugs having a 5-HT(1A) affinity is the presence of an arylpiperazine moiety. Indeed, the protonated nitrogen and the aromatic ring of the arylpiperazine compounds are considered crucial for the interaction with the receptor. However, the replacement of the piperazine moiety by a 1,2,3,6 tetrahydropyridine ring in 4-arylpiperazine-ethyl carboxamide derivatives was recently shown to be highly favourable for 5-HT(1A) affinity. In order to better understand the favourable effect of this chemical modification, we performed a conformational analysis of these compounds mainly based on the position of the phenyl ring relative to the piperazine and tetrahydropyridine moiety. In the piperazine compounds, the phenyl ring preferentially adopts a perpendicular orientation, whereas an almost planar orientation seems to be the most favourable conformation for the tetrahydropyridine compounds. Therefore, this conformational difference appears as a key for a better interaction with the receptor binding site. This result will serve for the designing high-affinity 5-HT(1A) ligands. PMID- 20022749 TI - Novel selective thiazoleacetic acids as CRTH2 antagonists developed from in silico derived hits. Part 2. AB - Structure-activity relationships have been established by exploring the eastern and western side of 5-thiazolyleacetic acids as CRTH2 (chemoattractant receptor homologous molecule expressed on Th2 cells) antagonists. Benzhydryl motifs in the 2-position of the thiazole was found to be most advantageous. The 4-thiazole position should either carry 3- or 4-fluorophenyl rings or a 4-pyridyl suitably substituted in the flanking 2-position. Several compounds with single digit nanomolar binding affinity and full antagonistic efficacy for human CRTH2 receptor were obtained. The compound series display a good PK profile and selectivity over a large number of other targets. PMID- 20022750 TI - Hybrid anticancer agents: isothiocyanate-progesterone conjugates as chemotherapeutic agents and insights into their cytotoxicities. AB - New hybrid molecules of isothiocyanate and progesterone and their metal complexes were synthesized exhibiting promising anti-proliferative and pro-apoptotic activity against breast and prostate cancer cells. These metal complex compounds exploited an existing cellular transport pathway for delivery of cytotoxic isothiocyanate moiety across cell membrane resulting in the inhibition of cell viability and inducing apoptosis. The highest apoptotic action was shown by the copper complex, which was mediated through the inhibition of Akt signaling similar to the one shown by isothiocyanate compounds. Our results underscore the possible role of metal redox cycling, and thus it is likely will open newer avenues for further optimization for the synthesis of novel active compounds through appropriate isothiocyanate pharmacophores. PMID- 20022751 TI - Label-free fluorescent real-time monitoring of adenylyl cyclase. AB - In cellular signaling, adenylyl cyclase plays a key role in the hydrolysis of ATP to cyclic AMP and pyrophosphate. Using a synthetic fluorescent chemosensor (PyDPA) which binds strongly to the pyrophosphate group, we have developed a label-free fluorescent real-time detection system for adenylyl cyclase. This assay would be the first adenylyl cyclase assay based on chemosensing the production of pyrophosphate. PMID- 20022752 TI - Tricyclic dihydroquinazolinones as novel 5-HT2C selective and orally efficacious anti-obesity agents. AB - Agonists of the 5-HT(2C) receptor have been shown to suppress appetite and reduce body weight in animal models as well as in humans. However, agonism of the related 5-HT(2B) receptor has been associated with valvular heart disease. Synthesis and biological evaluation of a series of novel and highly selective dihydroquinazolinone-derived 5-HT(2C) agonists with no detectable agonism of the 5-HT(2B) receptor is described. Among these, compounds (+)-2a and (+)-3c were identified as potent and highly selective agonists which exhibited weight loss in a rat model upon oral dosing. PMID- 20022753 TI - The antimicrobial activity of compounds from the leaf and stem of Vitis amurensis against two oral pathogens. AB - Nine compounds isolated from the leaf and stem of Vitis amurensis Rupr. (Vitaceae) were evaluated for their antimicrobial activity against two oral pathogens, Streptococcus mutans and Streptococcus sanguis, which are associated with caries and periodontal disease, respectively. The results of several antimicrobial tests, including MIC, MBC, and TBAI, showed that three compounds inhibited the growth of the test bacteria at concentrations ranging from 12.5 to 50 microg/mL. Among these compounds, compound 5, trans-epsilon-viniferin, displayed the strongest activity against S. mutans and S. sanguis with MIC values of 25 and 12.5 microg/mL, respectively. This is the first report on the antimicrobial activity of stilbenes and oligostilbenes isolated from the leaf and stem of V. amurensis. Thus, this result suggests that natural antimicrobial compounds derived from V. amurensis may benefit oral health as plaque-control agents for the prevention of dental caries and periodontal disease. PMID- 20022754 TI - Effects of a secondary task on postural control in children with Tourette syndrome. AB - Tourette syndrome (TS) is a neurodevelopmental disorder characterized by involuntary motor and vocal tics. Sub-clinical postural control anomalies have recently been reported in children with TS. The goal of the present study was to determine whether these anomalies interact with attention in postural control. Thirty-two younger (below 10 years) and 21 older (above 10 years) children with TS were compared to 13 younger and 15 older age-matched controls. Postural control was examined during standing with and without a secondary visual attention task. Sway velocity was higher in younger children than older ones and also higher in children with TS than in controls. The secondary task exacerbated the velocity anomalies in younger children with TS. The effects were independent of tic severity, medication, and attention deficit. The results suggest that postural control anomalies in TS are sensitive to attentional requirements. PMID- 20022757 TI - Comparison of triple-to-double coincidence ratio (TDCR) efficiency calculations and uncertainty assessments for 99Tc. AB - A comparison exercise for data analysis was recently conducted by the Liquid Scintillation Counting Working Group (LSCWG) of the International Committee on Radionuclide Metrology (ICRM) to evaluate the uncertainties involved in applying different analysis methodologies (including computer programs) for the triple-to double coincidence ratio (TDCR) method. The goals of the comparison were to (1) study differences in calculation results from different TDCR analysis programs, (2) investigate differences in analysis techniques and uncertainty assessment philosophies between laboratories, and (3) study the effect of not taking asymmetry of photomultiplier tube (PMT) efficiencies into account on the calculated activity. To achieve this, a single set of TDCR data for the pure beta emitter (99)Tc, was distributed to the participants, who analyzed the data according to their normal procedures and report the activity concentration of the (99)Tc solution from their results. The results indicate that the presently used programs are generally able to calculate the same activity values, assuming that the correct input parameters are used and that not taking PMT asymmetry into account in the calculations can lead to significant (0.6% for (99)Tc) errors in reported results. The comparison also highlighted the need for a more rigorous approach to estimating and reporting uncertainties. PMID- 20022755 TI - How antibodies fold. AB - B cells use unconventional strategies for the production of a seemingly unlimited number of antibodies from a very limited amount of DNA. These methods dramatically increase the likelihood of producing proteins that cannot fold or assemble appropriately. B cells are therefore particularly dependent on 'quality control' mechanisms to oversee antibody production. Recent in vitro experiments demonstrate that Ig domains have evolved diverse folding strategies ranging from robust spontaneous folding to intrinsically disordered domains that require assembly with their partner domains to fold; in vivo experiments reveal that these different folding characteristics form the basis for cellular checkpoints in Ig transport. Taken together, these reports provide a detailed understanding of how B cells monitor and ensure the functional fidelity of Ig proteins. PMID- 20022756 TI - A simple method for determination of natural and depleted uranium in surface soil samples. AB - A simple and efficient method for determination of uranium content in surface soil samples contaminated with depleted uranium, by gamma ray spectrometry is presented. The content of natural uranium and depleted uranium, as well as the activity ratio (235)U/(238)U of depleted uranium, were determined in contaminated surface soil samples by application of this method. PMID- 20022758 TI - Laboratory experiments to characterize radiochloride diffusion in unsaturated soils. AB - Diffusion transport of (36)Cl was examined in seven soils under unsaturated conditions in tubes packed with two portions of each soil having different (36)Cl activity concentrations. Apparent diffusion coefficients (D(a)) derived from diffusion profiles varied within a narrow range (from 3x10(-10) to 7x10(-10) m(2) s(-1)) confirming the minor effect of soil properties on the diffusion of a non reactive radionuclide like (36)Cl. Instead, packing conditions had a major effect. Solid-liquid distribution coefficients (K(d)) derived from D(a) (0.02-0.2 L kg(-1)) were systematically lower than those obtained from batch experiments (0.6-1.0 L kg(-1)), but with a similar variation pattern among soils. The low values of K(d) (Cl) confirmed an almost negligible radiochloride-soil interaction. PMID- 20022759 TI - Endobronchial ultrasound-guided needle aspiration in non-small-cell lung cancer restaging verified by the transcervical bilateral extended mediastinal lymphadenectomy--a prospective study. AB - OBJECTIVES: The aim of the study was to assess the diagnostic yield of the endobronchial ultrasound-guided transbronchial needle aspiration (EBUS-TBNA) in restaging of the non-small-cell lung cancer (NSCLC) patients after neo-adjuvant therapy. METHODS: In a consecutive group of NSCLC patients with pathologically confirmed N2 disease, who underwent neo-adjuvant chemotherapy, EBUS-TBNA was performed. All patients with negative EBUS-TBNA underwent subsequently the transcervical extended bilateral mediastinal lymphadenectomy (TEMLA) as a confirmatory test. RESULTS: A total of 61 patients underwent restaging EBUS-TBNA between 1 June 2007 and 31 December 2008. There were 85 mediastinal lymph nodes biopsied (stations: 2R - 2, 4R - 24, 2L - 1, 4L - 18 and 7 - 40). EBUS-TBNA revealed metastatic lymph node involvement in 18 of 61 patients (30%) and in 22 of 85 biopsies (26%). In 43 patients with negative or uncertain EBUS-TBNA, who underwent subsequent TEMLA, metastatic nodes were diagnosed in nine patients (15%) - in seven (12%) in stations accessible for EBUS-TBNA (stations: 2R - 1, 4R - 5, 7 - 4) and in two (3%) in station not accessible for EBUS-TBNA (station: 5 - 2). The false-negative results of biopsies were found only in small nodes (5.8+/ 2.8 mm x 7.5+/-2 mm). Moreover, all positive N2 nodes diagnosed by TEMLA contained only small metastatic deposits. There were three of 61 (5%) patients with false-positive results of biopsies in stations: 4R - 1, 4L - 1, and 7 - 2. A diagnostic sensitivity, specificity, accuracy, positive predictive value (PPV) and negative predictive value (NPV) of the restaging EBUS-TBNA was 67% (95% confidence interval (CI) - 65-90), 86% (95% CI - 82-95), 80%, 91% (95% CI - 80 100) and 78% (95% CI - 73-93), respectively. No complications of EBUS-TBNA were observed. CONCLUSIONS: EBUS-TBNA is an effective and safe technique for mediastinal restaging in NSCLC patients, and after the data presented in our study, in patients with negative results of EBUS-TBNA, a surgical restaging of the mediastinum might not be mandatory. PMID- 20022760 TI - Long-term results after resection for bone sarcoma pulmonary metastases. AB - BACKGROUND: Pulmonary metastases from bone sarcomas occur in approximately 40% of the cases. The combination of both chemotherapy and surgical resection is currently the standard treatment options for these patients. We aim to study the influence of different prognostic factors on long-term survival. METHODS: We reviewed the prognostic factors and survival rate in 52 consecutive patients with pulmonary metastases from bone sarcomas. All of them were previously treated with chemotherapy and submitted to metastasectomy at our institution from 1996 to 2006. Clinical and demographic variables, related to the primary tumour as well as to the pulmonary metastases, and treatment procedures were registered. Univariate (log-rank) and multivariate (Cox regression) analysis were carried out to identify significant prognostic factors related to overall survival. Five-year survival rates were estimated using Kaplan-Meier methods. RESULTS: Median follow up was 28 months. Follow-up duration ranged 7-148 months; the median survival was 27 months. As many as 31% of the patients were alive without disease, 3% were alive with disease, 64% died of disease while 2% died from other causes. Complete resection was achieved in 49 cases (94%). The overall 3- and 5-year survival rates were 43% and 31%, respectively. Univariate analysis showed (1) disease-free interval between treatment of the primary bone tumour and first lung metastasectomy (DFI) and (2) disease-free interval between first and second lung surgery (DFI2) as prognostic factors. Gender, primary site, histology of primary tumour, surgical approach, number of lung nodules, type of lung resection and re do lung surgery did not have a significant impact on survival. CONCLUSION: The long-term survival after bone sarcoma lung metastasectomy is encouraging. In our series, DFI and DFI2 were identified as the only prognostic factors. PMID- 20022761 TI - A combined approach of endobronchial and endoscopic ultrasound-guided needle aspiration in the radiologically normal mediastinum in non-small-cell lung cancer staging--a prospective trial. AB - OBJECTIVES: This prospective study aimed to assess the diagnostic yield of the combined approach - endobronchial (EBUS) and endoscopic (EUS) ultrasound-guided needle aspiration (combined ultrasound-needle aspiration (CUS-NA)) in the radiologically normal mediastinum in non-small-cell lung cancer (NSCLC) staging. METHODS: CUS-NA was performed simultaneously under local anaesthesia and sedation in consecutive NSCLC patients with mediastinal nodes that were not enlarged on CT (stage IA-IIB). All patients with negative CUS-NA subsequently underwent the transcervical extended bilateral mediastinal lymphadenectomy (TEMLA) as a confirmatory test. RESULTS: A total of 120 NSCLC patients underwent CUS-NA between 1 January 2008 and 31 December 2008. There were 318 mediastinal nodes biopsied (158 EBUS-NA - stations: 2R - 2, 2L - 1, 4R - 34, 4L - 33 and 7 - 88 and 160 EUS-NA - stations: 4L - 57, 7 - 101 and 9 - 2). CUS-NA revealed metastatic lymph node involvement in 19 of 120 patients (16%) and in 31 of 318 biopsies (10%). The prevalence was 22%. In 99 patients with negative CUS-NA, who underwent subsequent TEMLA, metastatic nodes were diagnosed in nine patients (8%) in 11 stations: 2R - 2, 4R - 4, 4L - 1, 5 - 3 and 7 - 1. In all but one patient there were 'minimal N2' only. Diagnostic sensitivity, specificity, total accuracy, positive predictive value (PPV) and negative predictive value (NPV) of CUS-NA for normal mediastinum was 68% (95% confidence interval (CI): 48-84), 98% (95% CI: 92 100), 91% (95% CI: 86-96), 91% (95% CI: 70-99) and 91% (95% CI: 83-96), respectively. The sensitivity of CUS-NA was significantly higher than with EBUS NA alone (p=0.04) and higher, close to the level of significance than with EUS-NA alone (p=0.07). The NPV of all techniques was high and that of CUS-NA was significantly higher than EBUS-NA alone and EUS-NA alone (p=0.01, p=0.03). No complications of CUS-NA were observed. CONCLUSIONS: In the radiologically normal mediastinum, CUS-NA is a highly effective and safe technique in NSCLC staging and, if negative, a surgical diagnostic exploration of the mediastinum may be omitted. PMID- 20022762 TI - Highly diversified innate receptor systems and new forms of animal immunity. AB - Detailed understanding of animal immunity derives almost entirely from investigations of vertebrates, with a smaller, but significant, contribution from studies in fruit flies. This limited phylogenetic scope has artificially polarized the larger view of animal immunity toward the complex adaptive immune systems of vertebrates on the one hand and systems driven by relatively small, stable families of innate receptors of insects on the other. In the past few years analyses of a series of invertebrate deuterostome genome sequences, including those from echinoderms and cephalochordates, sharply modify this view. These findings have far-reaching implications for characterizing the potential range of animal immunity and for inferring the evolutionary pathway that led to vertebrate immune systems. PMID- 20022763 TI - Radiofrequency ablation of small lung metastases by a single application of a 2 cm expandable electrode: determination of favorable responders. AB - PURPOSE: To determine which lung metastases are most likely to be treated effectively with a single radiofrequency (RF) application (defined as two separate applications of RF energy at a single electrode position) with a multitined expandable electrode with arrays 2 cm in diameter. MATERIALS AND METHODS: The authors retrospectively evaluated 88 lung metastases (mean long-axis diameter, 0.9 cm) in 36 patients (20 men and 16 women; mean age, 57 years) treated with a single RF application with a multitined expandable electrode with arrays 2 cm in diameter. Based on follow-up computed tomographic examinations, the technique effectiveness rates were estimated with Kaplan-Meier analysis. Multiple variables were analyzed with the log-rank test to determine risk factors for local progression. Then, the technique effectiveness rates were again estimated when considering only metastases without risk factors and compared with those of other tumors. RESULTS: The median follow-up period was 13.2 months (range, 6.0-24.7 months). Tumor size greater than 1.0 cm (P = .033) and contact with the bronchus with an inner diameter of at least 2 mm (P = .047) were the significant risk factors for local progression. The technique effectiveness rates for metastases 1.0 cm or smaller that were not in contact with the bronchus (n = 59) were 96% at 1 year and at 2 years; those rates were significantly (P = .010) higher than those in other tumors (n = 29). CONCLUSIONS: A single RF application with a multitined expandable electrode with arrays 2 cm in diameter is most likely to suffice in small (C mutation. Cohort 1A contains 82 patients [59 males, 23 females] of whom 42% are known to have cardiac complications of FD and 38% renal complications. Cohort 1B at the time of writing contained 37 patients [15 males, 22 females] of whom the indications for ERT were cardiac in 55% and renal in 60%. Cohort 1C at the time of writing contained 125 patients [22 males, 103 females]. Enrolment is ongoing in both Cohorts 1B and 1C. When compared to subjects in the Fabry Outcome Survey and the Fabry Registry, subjects in the CFDI are less likely to be male reflecting less ascertainment bias. The CFDI is a robust national data set that will contribute to available data on the natural history of FD and on the comparative efficacy of the two commercially available ERT products. PMID- 20022778 TI - Rotavirus disease burden, Nicaragua 2001-2005: defining the potential impact of a rotavirus vaccination program. AB - BACKGROUND: In October 2006, a rotavirus vaccine was introduced in Nicaragua for routine immunization of all children. We document the baseline diarrheal disease burden in Nicaragua prior to the vaccine program to facilitate future studies to measure vaccine impact. METHODS: We analyzed national data for 2001-2005 on total acute gastroenteritis healthcare visits, hospitalizations, and mortality in Nicaraguan children aged <5 years. RESULTS: Prior to vaccine introduction, by age 5 years, one in four Nicaraguan children required an outpatient consultation, one in 34 were hospitalized, and one in 2487 died from rotavirus-associated diarrhea, representing approximately 41,122 outpatient visits, 4460 hospitalizations, and 60 deaths per year that are preventable through vaccination. Almost half of the total acute gastroenteritis burden was in children <1 year of age. Two distinct seasonal peaks were noted in acute gastroenteritis hospitalizations and deaths. CONCLUSIONS: Existing data sources on all-cause acute gastroenteritis could be useful for establishing diarrhea disease burden and monitoring trends after vaccine introduction. Blunting of winter season peaks in rates of diarrhea, particularly among children aged <1-2 years, would be a useful indicator of impact from rotavirus vaccination. PMID- 20022779 TI - Relapse of severe sarcoidosis with an uncommon peritoneal location after TNFalpha blockade. Efficacy of rituximab, report of a single case. PMID- 20022781 TI - An atypical lombosciatica due to a sacral fracture revealing Crohn's disease. PMID- 20022782 TI - Liver impairment after concomitant administration of bosentan and clarithromycin in systemic sclerosis. PMID- 20022783 TI - Assessment of soluble cell adhesion molecules and soluble CD40 ligand levels in ankylosing spondylitis. PMID- 20022784 TI - Cartilage regeneration using adipose-derived stem cells and the controlled released hybrid microspheres. AB - OBJECTIVE: This study was to evaluate the effect of hybrid microspheres (MS) composed of gelatin transforming growth factor-beta (TGF-beta1)-loaded MS and chitosan MS on the enhancement of differentiation of adipose-derived stem cells (ASCs) into chondrocytes in pellet culture in vitro and the reparative capacity of pellet from ASCs and the hybrid MS-TGF used to repair cartilage defects in vivo. METHODS: The morphology of the controlled-released MS was observed with scanning electron microscopy (SEM) and mechanical property was also tested in this study. In vitro TGF-beta1 release was evaluated by an enzyme-linked immunosorbent assay. The protein expression of Collagen II was tested by Western blot. In addition, a preliminary study on cartilage regeneration was also performed in vivo. RESULTS: When chondrogenic differentiation of ASCs in both MS was evaluated, the protein expression of Collagen II became significantly increased for the hybrid MS-TGF, as compared with the gelatin MS-TGF. Mechanical result showed that the hybrid MS was superior to the gelatin MS. Observation of histology in vivo demonstrated that the pellet from ASCs and the hybrid MS-TGF promoted cartilage regeneration in the defects of articular cartilage much better than other groups. CONCLUSION: Our study demonstrated that the pellet from ASCs and the hybrid MS-TGF can provide an easy and effective way to construct the tissue engineered cartilage in vitro and in vivo. PMID- 20022785 TI - Effects of 14 weeks of TNF alpha blockade treatment on lipid profile in ankylosing spondylitis. AB - OBJECTIVE: Cardiovascular morbidity and mortality seem to be increased in ankylosing spondylitis, perhaps as the result of biological inflammation and consecutive dyslipidemia. This study aims to investigate the impact of TNF alpha inhibitors, an effective treatment, on lipid profile. METHODS: Thirty-four ankylosing spondylitis (AS) patients with active disease undergoing anti-TNF alpha therapy (n=20, infliximab; n=7, etanercept; n=7, adalimumab) were recruited. Disease activity parameters, total cholesterol, LDL-cholesterol, HDL cholesterol and triglycerides were assessed at baseline and after 14 weeks of treatment. RESULTS: After 14 weeks of TNF alpha blockade treatment, there was a significant increase in levels of total cholesterol (5.08+/-1.20 vs. 4.73+/-1.12 mmol/l; p=0.01) and HDL-cholesterol (1.61+/-0.47 vs. 1.47+/-0.35 mmol/l; p=0.008), but no resulting change in the atherogenic index (3.43+/-1.13 vs. 3.35+/-0.93; p=0.87). There was also no change in concentrations of triglycerides (1.33+/-1.22 vs. 1.27+/-0.98 mmol/l; p=0.794) and LDL-cholesterol (3.15+/-0.99 vs. 2.91+/-0.93 mmol/l; p=0.24). TNF alpha inhibitor treatment was followed by a significant improvement in all disease activity parameters: VAS pain or VAS disease activity, BASDAI or BASFI and systemic inflammation. Sub-group analysis showed that monoclonal antibodies increased total and LDL-cholesterol levels but did not change the atherogenic index. Conversely, 14 weeks of etanercept treatment was followed by no change in lipid profile. CONCLUSION: TNF alpha inhibitors may be successful in reducing cardiovascular risk in AS, as in RA, but not by affecting lipid profile. However, there is insufficient documented evidence, and long-term investigations are needed to define the possible protective mechanisms of TNFalpha inhibitor treatment in spondylarthropathies. PMID- 20022786 TI - Irrelevance of the WHO analgesic ladder for managing rheumatic pain. PMID- 20022787 TI - Childhood scleromyositis with a negative PM/Scl antibody. AB - Overlap syndromes, such as scleromyositis, antisynthetase syndrome and mixed connective tissue disease, consist of a blend of clinical features involved in two separate autoimmune disorders. Scleromyositis, the most common overlap syndrome, is usually observed in adults, and has rarely been documented in childhood. Here, the case of a 10-year-old African American female with scleromyositis is reported. This disease has features of both dermatomyositis and systemic scleroderma. Her disease process began with signs and symptoms of dermatomyositis and slowly changed clinically to features of systemic scleroderma. This patient had a positive ANA with a nucleolar pattern and a negative anti-PM/Scl antibody, an antibody which has been used as a prognostic and diagnostic marker of scleromyositis. PMID- 20022788 TI - Why all systemic lupus erythematosus patients should be given hydroxychloroquine treatment? PMID- 20022789 TI - [Residency in obstetrics and gynaecology in Alsace (France)]. PMID- 20022790 TI - [Assisted reproductive technologies for serodiscordant couples: desire for child and pregnancy versus the fact of illness]. AB - Assisted Reproductive Technologies for serodiscordant couples with HIV-infected male partner has been possible since 2000 in French centres such as the CECOS in Toulouse. From the outset, couples in the program meet psychologists or psychiatrists. In the first instance, the results from 40 psychologist led interviews with serodiscordant couples held at the CECOS in Toulouse, identified several challenging ideas. Couples who plan to have children were in a stable long-term relationship and both were involved in managing the illness. Medically assisted procreation awakens in the women a strong desire for children and enables the couples to make plans for the future. But it also reminds the men of their reliance on medical assistance because of the demands made by the medical program, and in case of failure it can destabilise the couples' way of coping with HIV. Subsequently, interviews with 12 couples during pregnancy revealed the overbearing presence of the illness, despite the imminent birth of the desired child. In their responses, women express their anxiety concerning the risk of contamination, and the defences that the men had built up to cope with HIV appeared destabilised, now that the idea of their death is rekindled. PMID- 20022791 TI - [Which hormones promote breast cancer in postmenopause: estrogens, progestins, insulin and/or adipocytokines?]. PMID- 20022793 TI - [Choose a fetal biometric reference curve using Z-scores]. PMID- 20022794 TI - [Lymph node surgical staging for locally advanced cervical cancer]. AB - Lymph node staging in patients with locally advanced cervical cancer is the most important prognostic factor and also leads to adjuvant treatment choice. Because of the lymphadenectomy associated morbidity and delay in the beginning of adjuvant therapy, noninvasive approaches were developed during the last decennia. Recently, positron emission tomography employing a glucose analogue (FDG-PET) has been shown to be more sensitive and more specific than magnetic resonance imaging or than computed tomography usually used in diagnosis of pelvic and para-aortic lymph node metastases. Even if recent studies have reported promising results, surgical pelvic and para-aortic staging remains actually the most accurate procedure for evaluating lymph node metastases. This procedure should be accomplished by transperitoneal or extraperitoneal laparoscopy, with the benefits of minimal morbidity, shorter length of hospital stay and no significant increase of complications comparing to laparotomy approach. Laparoscopy also allows an early start of adjuvant treatment, this delay constituting an important prognostic factor for patients with locally advanced cancer. However, the survival benefit of lymph node dissection is still controversial and should be proved in randomised studies. PMID- 20022795 TI - [Women in obstetrics and gynaecology: an association with a future]. PMID- 20022796 TI - Cardiac flow component analysis. AB - In a chamber of the heart, large-scale vortices are shown to exist as the result of the dynamic blood flow and unique morphological changes of the chamber wall. As the cardiovascular flow varies over a cardiac cycle, there is a need for a robust quantification method to analyze its vorticity and circulation. We attempt to measure vortex characteristics by means of two-dimensional vorticity maps and vortex circulation. First, we develop vortex component analysis by segmenting the vortices using an data clustering algorithm before histograms of their vorticity distribution are generated. The next stage is to generate the statistics of the vorticity maps for each phase of the cardiac cycle to allow analysis of the flow. This is followed by evaluating the circulation of each segmented vortex. The proposed approach is dedicated to examining vortices within the human heart chamber. The vorticity field can indicate the strength and number of large-scale vortices in the chamber. We provide the results of the flow analysis after vorticity map segmentation and the statistical properties that characterize the vorticity components. The success of the cardiac measurement and analysis is illustrated by a case study of the right atrium. Our investigation shows that it is possible to utilize a data clustering algorithm to segment vortices after vorticity mapping, and that the vorticity and circulation analysis of a chamber vorticity can provide new insights into the blood flow within the cardiovascular structure. PMID- 20022797 TI - The resolution of inflammation and cancer. AB - Inflammation has long been thought to contribute to the development of cancer; however there is also clear evidence that the immune system can recognize and eliminate cancer cells. Current research suggests that cancer-associated inflammation has a dual role in tumor progression; inflammatory mediators promote the malignant activity of cancer cells by acting as growth factors and also stimulate angiogenesis, however, cancer-associated inflammation is also linked with immune-suppression that allows cancer cells to evade detection by the immune system. In this review we will discuss the dual role of inflammation in cancer and how endogenous anti-inflammatory mechanisms may equally be important in carcinogenesis. PMID- 20022798 TI - A broadening world of bacterial small RNAs. AB - The ubiquity of small RNAs (sRNAs) in bacteria is now well established. These transcripts are the members of regulatory circuits involved in diverse processes ranging from stress adaptation to virulence to metabolism. Recent large-scale searches suggest that there exist many times more sRNAs than previously predicted even in the best studied bacterial transcriptomes. On the basis of these and other recent findings of regulatory sRNAs that do not function in a 'classical' manner, we propose that the working definition of sRNAs be broadened. PMID- 20022799 TI - Finding a way to the nucleus. AB - Agrobacterium species transfer single-strand DNA and virulence effector proteins to plants. To understand how Agrobacterium achieves interkingdom horizontal gene transfer, scientists have investigated how the interaction of bacterial effector proteins with host proteins directs T-DNA to the plant nucleus. VirE2, a single strand DNA binding protein, likely plays a key role in T-DNA nuclear targeting. However, subcellular trafficking of VirE2 remains controversial, with reports of both cytoplasmic and nuclear localization. The recent discovery that phosphorylation of the VirE2 interacting protein VIP1 modulates both nuclear targeting and transformation may provide a solution to this conundrum. Novel experimental systems that allow tracking of VirE2 as it exits Agrobacterium and enters the plant cell will also aid in understanding virulence protein/T-DNA cytoplasmic trafficking. PMID- 20022800 TI - Fluorescence and DNA-binding spectral studies of neodymium(III) complex containing 2,2'-bipyridine, [Nd(bpy)2Cl(3)xOH2]. AB - The interaction of [Nd(bpy)(2)Cl(3)xOH(2)], where bipy is 2,2'-bipyridine, with DNA has been studied by absorption, emission, and viscosity measurements. [Nd(bpy)(2)Cl(3)xOH(2)] showed absorption decreasing in charge transfer band with increasing of DNA. The binding constant, K(b) has been determined by absorption measurement and found to be (1.5+/-0.1)x10(5)M(-1). The fluorescent of [Nd(bpy)(2)Cl(3)xOH(2)] has been investigated in detail. The interaction was also studied by fluorescence quenching technique. The results of fluorescence titration revealed that DNA had the strong ability to quenching the intrinsic fluorescence of Nd(III) complex at 327 nm. The binding site number n, apparent binding constant K(b) and the Stern-Volmer quenching constant K(SV) have been determined. Thermodynamic parameters have been calculated according to relevant fluorescent data and Van't Hoff equation. Characterization of bonding mode has been studied. The results suggested that the major interaction mode between [Nd(bpy)(2)Cl(3)xOH(2)] and DNA was groove binding. PMID- 20022801 TI - Influence of metal ions on the interaction between gatifloxacin and calf thymus DNA. AB - To study the interaction between gatifloxacin (GT), metal ions (Cu(2+), Cd(2+), Co(2+), Mg(2+)) and calf thymus DNA under condition of physiology pH, UV absorption and fluorescence methods were adopted. Result shows that metal ions and DNA are able to react with GT in ground state. In further research, by studying the influence of metal ions on binding of GT with DNA in metal ions-GT DNA ternary system, we found that influential mechanism of Mg(2+) on the binding of GT with DNA may be different from the other three. Mg(2+) can act as a bridge in the binding of GT's carboxyl/carbonyl with DNA phosphate in certain concentration range; while Cu(2+), Cd(2+), Co(2+) can combine directly with GT by reaction between GT carboxyl/carbonyl and DNA base, and enhance the binding ability of GT with DNA. The influence extent and type depend not only on the binding site of DNA with metal ions (phosphate or base), but also the binding ability of which. The stronger the binding ability of metal ions with DNA base is, the larger their promotion to binding of GT with DNA is. The order of metal ions' influential ability on the binding of GT-DNA is identical to the binding ability order of metal ions with DNA base, that is: Cu(2+)>Cd(2+)>Co(2+)>Mg(2+). PMID- 20022802 TI - Experimental and theoretical study on vibrational spectra of nickel and zinc complexes of 5,10-diphenylporphyrin. AB - The vibrational spectra of nickel and zinc 5,10-diphenylporphyrin (NiDaPP and ZnDaPP) have been studied by density functional theory (B3LYP/6-31G(d)) and experimental measurement. The assignment of observed Raman and IR bands are proposed based on theoretical calculation. Theoretical study indicates that 5,10 diphenyl substitution together with out-of-plane distortion diversify the structure of four pyrrole rings and the environment around C(m) atoms, which lowers the symmetry of porphyrin skeleton and brings about some variation to vibrational spectra. The first is the activation of all normal modes both in Raman and IR spectra. The second is the lifting of original degenerate E(u) representation and its splitting into two A modes in NiDaPP (C(1) group point) and A'/A'' presentation in ZnDaPP (C(s) group point), respectively. Furthermore, one or both of the two split components, especially for some vibration involving the motion of C(m) atom, undergoes further mixing with original non-degenerate mode (A(1g), A(2g), B(1g), or B(2g)) of same vibration. This produces new modes involving only partial motion of original mode. Besides general increasing frequency of NiDaPP with respect to ZnDaPP, different vibrational structure is also indicated by calculation for some structure-sensitive bands, especially nu(8) mode that couples with gamma(6) mode and split into two modes for NiDaPP but not for ZnDaPP. PMID- 20022803 TI - Net analyte signal standard addition method (NASSAM) as a novel spectrofluorimetric and spectrophotometric technique for simultaneous determination, application to assay of melatonin and pyridoxine. AB - In this work a new modification of the standard addition method called "net analyte signal standard addition method (NASSAM)" is presented for the simultaneous spectrofluorimetric and spectrophotometric analysis. The proposed method combines the advantages of standard addition method with those of net analyte signal concept. The method can be applied for the determination of analyte in the presence of known interferents. The accuracy of the predictions against H-point standard addition method is not dependent on the shape of the analyte and interferent spectra. The method was successfully applied to simultaneous spectrofluorimetric and spectrophotometric determination of pyridoxine (PY) and melatonin (MT) in synthetic mixtures and in a pharmaceutical formulation. PMID- 20022804 TI - Relevance of HPV mRNA detection in a population of ASCUS plus women using the NucliSENS EasyQ HPV assay. AB - BACKGROUND: DNA- and mRNA-based assays are the main tools used for detecting human papillomavirus (HPV) nucleic acid in clinical samples. A recent tool, NucliSENS EasyQ HPV, uses a new concept to directly detect the expression of HPV oncogenic factors (E6 and E7) from the most prevalent HPV genotypes in cervical cancer (16, 18, 31, 33 and 45). OBJECTIVES: The primary aim of the study is to assess the accuracy of NucliSENS EasyQ HPV in detecting high-risk (HR) HPV in a population of atypical cells of undetermined significance/low-grade squamous intraepithelial lesion/high-grade squamous lesion (ASCUS/LSIL/HSIL) patients using a clinical cut-off of a cervical dysplasia (CIN2+) histology. The secondary aim is to compare this mRNA-based assay with the DNA-based hybrid capture II (HCII) assay. STUDY DESIGN: The study population comprised 140 women referred for colposcopy and histology. NucliSENS EasyQ HPV test, hybrid capture II (HCII) test and linear array (LA) test were assessed on all samples. All the tests were performed on the samples collected in PreservCyt liquid media for liquid-based cytology (ThinPrep Pap test). RESULTS: The clinical specificity of the NucliSENS EasyQ HPV was 63% for the detection of CIN2+ or HSIL patients, significantly higher than the specificity of HCII and LA (49% and 45%, respectively, p<0.05). Agreement between HCII and NucliSENS EasyQ HPV was fair (k=0.49) and was good between HCII and LA (k=0.88). HPV 16 was the most-detected type (49% with NucliSENS EasyQ HPV and 56% with LA), and HPV 31 was the second most-detected HPV type (31% with NucliSENS EasyQ HPV and 29% with LA). CONCLUSIONS: The NucliSENS EasyQ HPV assay has interesting clinical sensitivity and specificity for the detection of HPV types in CIN2+ patients and shows comparable diagnostic values with the HCII DNA assay. This assay allows simultaneous detection of HPV mRNA and determination of the type of the main prevalent oncogenic virus. PMID- 20022805 TI - Differential diagnosis of unpleasant sensations in the legs: prevalence of restless legs syndrome in a primary care population. AB - BACKGROUND: Restless legs syndrome (RLS) is a common neurological condition. We investigated the prevalence of RLS in patients suffering from unpleasant sensations in the legs. METHODS: We included 16,543 patients consulting one of 312 primary care practices in Germany on November 8, 2007. All patients filled out a self-assessment questionnaire. Patients who reported suffering from unpleasant sensations in the legs were then assessed by the physician. Main outcome measures were the overall prevalence of unpleasant sensations in the legs and the prevalence of RLS; the most common differential diagnoses in the subpopulation suffered from unpleasant leg sensations. RESULTS: Out of all participating patients 7704 (46.6%) suffered from unpleasant sensations in the legs and 1758 (10.6%) were diagnosed with RLS according to the four essential clinical criteria. Among patients with unpleasant leg sensations, the prevalence of RLS was considerably higher (22.7%) than in the total population. The most common differential diagnoses were osteoarthritis (21.5%), disc lesion (19.2%), varicose veins (18.8%) and muscle cramps (14.6%). Of the patients with RLS 53.4% had already consulted their physician about their leg problems in the past. Still, only 20.1% of the RLS patients had received the correct diagnosis. Comorbidity rates were significantly increased in RLS patients compared to patients suffering from leg symptoms of other origin. CONCLUSIONS: This study showed a high prevalence of RLS in primary care patients with unpleasant sensations in the legs. Thus, in patients presenting with these symptoms the diagnosis of RLS should routinely be considered. PMID- 20022806 TI - Maintaining cell polarity through vegetative cell pattern dedifferentiation: cytoskeleton and morphogenesis in the hypotrich ciliate Sterkiella histriomuscorum. AB - The morphological differentiation of ciliates is achieved through the development of a submembraneous cytoskeleton in which the cilia are anchored. In most hypotrich ciliates, this cytoskeleton is mainly constructed of microtubules. In these species, cells pass through vegetative cell pattern dedifferentiated stages during their biological cycle. In order to investigate the behaviour of the cytoskeleton during these stages, we analysed the reorganization of the cytoskeleton during the sexual cycle of Sterkiella histriomuscorum by microscopy. Sterkiella exconjugants transiently dedifferentiate to form zygocysts devoid of ciliature and infraciliature. Immunofluorescence images obtained with antibodies directed against pericentrosomal material and tubulin showed that the cells resorb their ciliature and basal bodies, but retain their submembraneous microtubular cytoskeleton during the whole process and that the body plan is maintained through vegetative cell pattern dedifferentiation: the cell polarity remains printed on the cell surface by the microtubular cytoskeleton which in turn could mark the sites of basal body assembly during zygocyst morphogenesis. The results are discussed in terms of mechanisms of cell patterning. PMID- 20022807 TI - Identification of prophage gene z2389 in Escherichia coli EDL933 encoding a DNA cytosine methyltransferase for full protection of NotI sites. AB - Pulsed-field gel electrophoresis (PFGE) analysis revealed that the genomes of some pathogenic Escherichia coli O157:H7 strains, including EDL933, were resistant to NotI digestion. An amino acid sequence comparison suggested that the z2389 gene carried on prophage CP-933R in strain EDL933 is likely to encode a C(5)-cytosine methyltransferase. The z2389-equivalent gene was found in the NotI resistant strains tested, but it was not detected in the NotI-susceptible strains. PFGE analysis of the wild-type EDL933 strain and of a z2389 null mutant revealed that z2389 was associated with full genome protection against NotI digestion and partial protection against EagI digestion. In vitro methylation experiments with purified recombinant protein demonstrated that Z2389 is capable of methylating NotI and EagI sites. Sequencing of bisulfite-treated DNA indicated that the methylation occurred at the first cytosine residue of the NotI recognition sequence, whereas EagI sites remained unmethylated or were methylated at the first cytosine residue. Thus, z2389 encodes a DNA cytosine methyltransferase that confers full protection to NotI sites. PMID- 20022808 TI - Drug-eluting stents for the treatment of acute myocardial infarction: the view to the HORIZONS. AB - Drug-eluting stents (DES) offer an attractive option for the treatment of acute thrombotic lesions during acute ST-elevation myocardial infarction (STEMI) due to their ability to inhibit restenosis. Several randomised trials have demonstrated the efficacy of DES in reducing target vessel revascularisation (TVR) in this setting. However, several registries of real-world patients receiving DES for STEMI have raised long-term safety concerns about DES use in this patient subset. Given the inherent limitations of registry data, this issue is likely to remain unresolved until further data is made available from large-scale ongoing trials with long-term follow-up such as the HORIZONS-AMI trial. PMID- 20022809 TI - Gefitinib versus cisplatin plus docetaxel in patients with non-small-cell lung cancer harbouring mutations of the epidermal growth factor receptor (WJTOG3405): an open label, randomised phase 3 trial. AB - BACKGROUND: Patients with non-small-cell lung cancer harbouring mutations in the epidermal growth factor receptor (EGFR) gene respond well to the EGFR-specific tyrosine kinase inhibitor gefitinib. However, whether gefitinib is better than standard platinum doublet chemotherapy in patients selected by EGFR mutation is uncertain. METHODS: We did an open label, phase 3 study (WJTOG3405) with recruitment between March 31, 2006, and June 22, 2009, at 36 centres in Japan. 177 chemotherapy-naive patients aged 75 years or younger and diagnosed with stage IIIB/IV non-small-cell lung cancer or postoperative recurrence harbouring EGFR mutations (either the exon 19 deletion or L858R point mutation) were randomly assigned, using a minimisation technique, to receive either gefitinib (250 mg/day orally; n=88) or cisplatin (80 mg/m(2), intravenously) plus docetaxel (60 mg/m(2), intravenously; n=89), administered every 21 days for three to six cycles. The primary endpoint was progression-free survival. Survival analysis was done with the modified intention-to-treat population. This study is registered with UMIN (University Hospital Medical Information Network in Japan), number 000000539. FINDINGS: Five patients were excluded (two patients were found to have thyroid and colon cancer after randomisation, one patient had an exon 18 mutation, one patient had insufficient consent, and one patient showed acute allergic reaction to docetaxel). Thus, 172 patients (86 in each group) were included in the survival analyses. The gefitinib group had significantly longer progression-free survival compared with the cisplatin plus docetaxel goup, with a median progression-free survival time of 9.2 months (95% CI 8.0-13.9) versus 6.3 months (5.8-7.8; HR 0.489, 95% CI 0.336-0.710, log-rank p<0.0001). Myelosuppression, alopecia, and fatigue were more frequent in the cisplatin plus docetaxel group, but skin toxicity, liver dysfunction, and diarrhoea were more frequent in the gefitinib group. Two patients in the gefitinib group developed interstitial lung disease (incidence 2.3%), one of whom died. INTERPRETATION: Patients with lung cancer who are selected by EGFR mutations have longer progression-free survival if they are treated with gefitinib than if they are treated with cisplatin plus docetaxel. FUNDING: West Japan Oncology Group (WJOG): a non-profit organisation supported by unrestricted donations from several pharmaceutical companies. PMID- 20022811 TI - Using microRNAs to understand cancer biology. PMID- 20022810 TI - Relation between microRNA expression and progression and prognosis of gastric cancer: a microRNA expression analysis. AB - BACKGROUND: Analyses of microRNA expression profiles have shown that many microRNAs are expressed aberrantly and correlate with tumorigenesis, progression, and prognosis of various haematological and solid tumours. We aimed to assess the relation between microRNA expression and progression and prognosis of gastric cancer. METHODS: 353 gastric samples from two independent subsets of patients from Japan were analysed by microRNA microarray. MicroRNA expression patterns were compared between non-tumour mucosa and cancer samples, graded by diffuse and intestinal histological types and by progression-related factors (eg, depth of invasion, metastasis, and stage). Disease outcome was calculated by multivariable regression analysis to establish whether microRNAs are independent prognostic factors. FINDINGS: In 160 paired samples of non-tumour mucosa and cancer, 22 microRNAs were upregulated and 13 were downregulated in gastric cancer; 292 (83%) samples were distinguished correctly by this signature. The two histological subtypes of gastric cancer showed different microRNA signatures: eight microRNAs were upregulated in diffuse-type and four in intestinal-type cancer. In the progression-related signature, miR-125b, miR-199a, and miR-100 were the most important microRNAs involved. Low expression of let-7g (hazard ratio 2.6 [95% CI 1.3-4.9]) and miR-433 (2.1 [1.1-3.9]) and high expression of miR-214 (2.4 [1.2 4.5]) were associated with unfavourable outcome in overall survival independent of clinical covariates, including depth of invasion, lymph-node metastasis, and stage. INTERPRETATION: MicroRNAs are expressed differentially in gastric cancers, and histological subtypes are characterised by specific microRNA signatures. Unique microRNAs are associated with progression and prognosis of gastric cancer. FUNDING: National Cancer Institute. PMID- 20022812 TI - Epigenetics: molecular mechanisms and implications for disease. AB - Epigenetics is rising to prominence in biology as a mechanism by which environmental factors have intermediate-term effects on gene expression without changing the underlying genetic sequence. This can occur through the selective methylation of DNA bases and modification of histones. There are wide-ranging implications for the gene-environment debate and epigenetic mechanisms are causing a reevaluation of many traditional concepts such as heritability. The reversible nature of epigenetics also provides plausible treatment or prevention prospects for diseases previously thought hard-coded into the genome. Here, we consider how growing knowledge of epigenetics is altering our understanding of biology and medicine, and its implications for future research. PMID- 20022813 TI - Objective evaluation of personality and psychopathology in temporal lobe versus extratemporal lobe epilepsy. AB - Previous research has been equivocal on personality trait and psychopathology differences between temporal lobe and other types of epilepsy, as well as between patients with right and left temporal lobe seizure foci. In this study, personality differences between patients with right temporal (n=23), left temporal (n=21), and extratemporal (n=24) epilepsy were investigated using the NEO Personality Inventory-Revised (NEO-PI-R). No statistically significant differences were found on any of the NEO-PI-R domains or facet trait scales. There were also no significant differences between groups on the Minnesota Multiphasic Personality Inventory 2 (MMPI-2), a measure of psychopathology. However, mild elevations were seen in all groups on clinical scales related to physical symptoms, health concern, and depression. These data suggest there are no consistent personality or psychopathology differences, as measured by the NEO PI-R and the MMPI-2, between patients with left temporal, right temporal, and extratemporal epilepsy whose seizures are localized using video/EEG monitoring. PMID- 20022815 TI - Funny channel-based pacemaking. PMID- 20022814 TI - Outcomes and complications of catheter ablation for atrial fibrillation in females. AB - BACKGROUND: Most atrial fibrillation (AF) ablation studies have consisted predominantly of males; accordingly, there is a paucity of information on the safety and efficacy of catheter ablation in a large cohort of female AF patients. OBJECTIVE: The purpose of this study was to evaluate catheter ablation for AF in female patients. METHODS: From January 2005 to May 2008, 3265 females underwent pulmonary vein antrum isolation. Success rates, patient profiles, and complications were collected. RESULTS: Approximately 16% of our population was female (P <.001). Females were older (59 +/- 13 vs. 56 +/- 19 years; P <.01) and had a lower prevalence of paroxysmal atrial fibrillation (PAF; 46% vs. 55%; P <.001). Females failed more antiarrhythmics (4 +/- 1 vs. 2 +/- 3; P = .04) and were referred later for catheter ablation (6.51 +/- 7 vs. 4.85 +/- 6.5 years; P = .02) than males. More females failed ablation (31.5% vs. 22.5%; P = .001) and had nonantral sites of firing than males (P <.001). Female patients had 11 (2.1%) hematomas versus 27 (0.9%) in males. CONCLUSIONS: Five times as many males underwent catheter ablation than females. Females failed more ablations possibly because of a higher prevalence of nonantral firing, non-PAF, and longer history of AF. Females had more bleeding complications than males. PMID- 20022816 TI - Cardiac I(to), KCNE2, and Brugada syndrome: promiscuous subunit interactions, or what happens in HEK cells stays in HEK cells? PMID- 20022817 TI - Atrial fibrillation and inflammation: quest for the missing link in the etiopathogenesis of AF. PMID- 20022818 TI - Utility of tachycardia cycle length variability in discriminating atrial tachycardia from ventricular tachycardia. AB - BACKGROUND: Inappropriate implantable cardioverter-defibrillator (ICD) therapy of atrial tachycardia (AT) with 1:1 atrioventricular (AV) conduction is common because it is difficult to discriminate from ventricular tachycardia (VT) with 1:1 retrograde conduction. Tachycardia cycle length (CL) variability and the relationship between atrial and ventricular CLs may be useful in discriminating AT from VT with 1:1 retrograde conduction. OBJECTIVE: The purpose of this study was to evaluate the usefulness of the relationship between the atrial and ventricular CLs in differentiating AT with 1:1 conduction from VT with 1:1 retrograde conduction. METHODS: We studied 71 patients who had a tachycardia with a 1:1 AV relationship and significant CL variability. Thirty-nine patients had AT (21 inducible and 18 simulated), and 32 patients had VT (11 inducible and 21 simulated). The relationship between atrial and ventricular CLs was examined. RESULTS: A change in atrial CL predicted the change in subsequent ventricular CL in 37 (95%) of 39 patients with AT and in none of the patients with VT. A change in preceding ventricular CL predicted the change in atrial CL in 31 (97%) of 32 patients with VT and in only one (3%) of 39 patients with AT. The sensitivity, specificity, and positive and negative predictive values of a change in atrial CL predicting the change in ventricular CL for AT with significant CL variability were 95%, 100%, 100%, and 94%, respectively. The corresponding values for the change in preceding ventricular CL predicting the change in atrial CL for AT with significant CL variability were 97%. CONCLUSION: The relationship between atrial and ventricular CL is useful in differentiating AT from VT with retrograde conduction. A change in atrial CL that predicts the change in subsequent ventricular CL rules in AT and excludes VT. PMID- 20022819 TI - Intracardiac and extracardiac markers of inflammation during atrial fibrillation. AB - BACKGROUND: A decrease in inflammation after cure of atrial arrhythmias suggests that such arrhythmias are proinflammatory, and lower inflammatory marker levels in the coronary sinus suggest that atrial arrhythmias result in intracardiac appropriation of inflammatory cytokines. OBJECTIVE: The purpose of this study was to investigate the effect of atrial fibrillation on inflammatory markers drawn from intracardiac and extracardiac chambers. METHODS: We performed a case-control study of 167 AF patients and 207 controls. Blood from intracardiac and extracardiac sites was obtained from a subset of patients undergoing curative AF ablation (n = 46). RESULTS: No significant differences in C-reactive protein (CRP) or interleukin-6 (IL-6) levels were seen between patients with and those without a history of AF. Both levels were significantly higher when blood was drawn during AF than during sinus rhythm: median CRP 3.1 mg/dL (interquartile range [IQR] 1.0-6.0) versus 1.7 mg/dL (IQR 0.7-3.9, P = .0005); median IL-6 2.3 ng/mL (IQR 1.5-3.9) versus 1.5 ng/mL (IQR 0.7-2.5, P = .007). This finding persisted after adjusting for potential confounders. AF ablation patients in AF exhibited a positive median left atrial minus coronary sinus gradient CRP (0.3 mg/dL, IQR -0.03-1.1), whereas those in sinus rhythm had a negative median left atrial minus coronary sinus gradient CRP (-0.2, IQR -0.8-[-0.02], P = .01). Femoral artery minus femoral vein gradients in AF versus sinus rhythm did not show any differences. CONCLUSION: AF at the time of the blood draw, rather than a history of AF, was independently associated with inflammation. Differences in transcardiac gradients suggest that AF results in sequestration of inflammatory cytokines in the heart. PMID- 20022820 TI - System survival of nontransvenous implantable cardioverter-defibrillators compared to transvenous implantable cardioverter-defibrillators in pediatric and congenital heart disease patients. AB - BACKGROUND: Nontransvenous (NTV) implantable cardioverter-defibrillator (ICD) systems with pericardial and/or subcutaneous coils are used in select pediatric and congenital heart disease patients who are not candidates for transvenous ICD leads. Outcomes with these hybrid configurations are not well understood. OBJECTIVE: The purpose of this study was to compare survival of NTV ICD systems to standard transvenous (TV) ICD systems. METHODS: We conducted a retrospective single-center study in which the TV group was matched to the NTV group 2:1 by type of cardiac disease and implant date. RESULTS: There were 39 patients in the NTV group and 78 matched in the TV group. Compared to the TV group, the NTV group was younger (median 7 vs 20 years) with a smaller body surface area at implant (0.9 vs 1.8 m(2); both P <.001). Median follow-up in the NTV group was 17 months. System survival at 12, 24, and 36 months was 73%, 55%, and 49% in the NTV group versus 91%, 83%, and 76% in the TV group (P = .003). A multivariable Cox proportional hazards model including group, body surface area, and age at implant revealed the NTV group to be an independent predictor of system failure (hazard ratio 2.9, P = .04). Rate of total unanticipated interventions in the NTV group was 18 versus 6 per 1,000 person-months in the TV group. In patients with NTV systems, 23% received appropriate shocks and 18% received inappropriate shocks. CONCLUSION: Survival of ICD systems using NTV defibrillation coils is significantly shorter than with TV ICD systems. Although NTV systems provide protection for this unique subset of patients, more durable options are needed. PMID- 20022821 TI - Mechanism of right precordial ST-segment elevation in structural heart disease: excitation failure by current-to-load mismatch. AB - BACKGROUND: The Brugada sign has been associated with mutations in SCN5A and with right ventricular structural abnormalities. Their role in the Brugada sign and the associated ventricular arrhythmias is unknown. OBJECTIVE: The purpose of this study was to delineate the role of structural abnormalities and sodium channel dysfunction in the Brugada sign. METHODS: Activation and repolarization characteristics of the explanted heart of a patient with a loss-of-function mutation in SCN5A (G752R) and dilated cardiomyopathy were determined after induction of right-sided ST-segment elevation by ajmaline. In addition, right ventricular structural discontinuities and sodium channel dysfunction were simulated in a computer model encompassing the heart and thorax. RESULTS: In the explanted heart, disappearance of local activation in unipolar electrograms at the basal right ventricular epicardium was followed by monophasic ST-segment elevation. The local origin of this phenomenon was confirmed by coaxial electrograms. Neither early repolarization nor late activation correlated with ST segment elevation. At sites of local ST-segment elevation, the subepicardium was interspersed with adipose tissue and contained more fibrous tissue than either the left ventricle or control hearts. In computer simulations entailing right ventricular structural discontinuities, reduction of sodium channel conductance or size of the gaps between introduced barriers resulted in subepicardial excitation failure or delayed activation by current-to-load mismatch and in the Brugada sign on the ECG. CONCLUSION: Right ventricular excitation failure and activation delay by current-to-load mismatch in the subepicardium can cause the Brugada sign. Therefore, current-to-load mismatch may underlie the ventricular arrhythmias in patients with the Brugada sign. PMID- 20022822 TI - High-performance ion chromatography method for separation and quantification of inositol phosphates in diets and digesta. AB - A gradient high-performance ion chromatographic method for separation and quantification of inositol phosphates (InsP(2)-InsP(6)) in feedstuffs, diets, gastric and ileal digesta from pigs was developed and validated. The InsP(2) InsP(6) were separated on a Dionex CarboPac PA1 column using a gradient with 1.5 mol L(-1) methanesulfonic acid and water. The exchange of the commonly used HCl with methanesulfonic acid has two advantages: (i) the obtained baseline during the separation is almost horizontal and (ii) it is not necessary to use an inert HPIC equipment as the methanesulfonic acid is not as aggressive as HCl. Twenty three of the 27 separated inositol phosphate isomers were isolated. ICP-MS was used for quantification of phosphorus in the isolated isomers and used for calculation of correction factors for each isomer allowing InsP(6) to be used as calibration standard. The detection limits for InsP(2)-InsP(6) were in the range of 0.9-4.4 mg phosphorus L(-1). The recovery of the major part of the inositol phosphates was 80-100%, and the CV for repeatability and reproducibility were 1 17% and 1-14%, respectively. PMID- 20022824 TI - [Physical preventive measures]. PMID- 20022823 TI - Descriptive epidemiology of cholangiocarcinoma in Italy. AB - BACKGROUND: Very little data exist on the epidemiology of cholangiocarcinoma in Italy. AIM: We focus on the descriptive epidemiology of cholangiocarcinoma in Italy. METHODS: Data on incidence were obtained from the Italian Association of Tumour Registries while mortality data were obtained from the Italian National Institute of Statistics. RESULTS: A progressive increase of incidence with age was seen for extra-hepatic, intra-hepatic and not otherwise specified cholangiocarcinoma. Crude incidence rates were higher for extra-hepatic cholangiocarcinoma than those for intra-hepatic cholangiocarcinoma and in men compared to women. An increasing incidence trend was observed, from 1988 to 2005, for both extra-hepatic- and intra-hepatic cholangiocarcinoma with a 3-6% yearly increase and with a rate of increase higher for men than for women and for intra hepatic- than for extra-hepatic cholangiocarcinoma. For intra-hepatic cholangiocarcinoma, the mortality rates progressively increased from 0.15 per million in 1980 to 5.9 per million in 2003, when mortality for this cancer surpassed extra-hepatic cholangiocarcinoma. Mortality rates for extra-hepatic cholangiocarcinoma showed an increasing trend from 1980 to 1994 but, in contrast to intra-hepatic cholangiocarcinoma, a stable or slightly decreasing trend from 1995 to 2003 was observed. CONCLUSIONS: In Italy, cholangiocarcinoma showed a progressive increase in incidence and mortality in the last two decades mainly in intra-hepatic cholangiocarcinoma. PMID- 20022825 TI - [Immunization against pandemic influenza]. PMID- 20022826 TI - [Methodologic aspects of the report on pandemic A (H1N1) influenza 2009]. PMID- 20022827 TI - [Infantile vascular tumors]. AB - The use in the past of an imprecise terminology to designate vascular tumors has contributed to its incorrect diagnosis, and as a consequence, to inadequate treatment. In childhood, different types of vascular tumors may be present. Hemangiomas of infancy are by far the most frequent, and other less common types are congenital hemangiomas (rapidly involuting or RICH and non-involuting or NICH), kaposiform hemangioendothelioma, angioblastoma or tufted angioma and pyogenic granuloma. The correct knowledge and diagnosis, always in a multidisciplinary setting, is required to reduce incorrect diagnosis, unnecessary complementary examinations and invasive tests, and for the patient to receive the most effective and precise treatment in each case. This article reviews the historical evolution, nomenclature and classification of vascular lesions, the different clinical and pathological characteristics of each vascular tumor, the complementary examinations required correct diagnosis, the differential diagnosis, as well as highlighting the treatment options currently available for different vascular tumors and related clinical conditions. PMID- 20022828 TI - [Magnitude and risk groups]. PMID- 20022829 TI - [Clinical and laboratory diagnosis]. PMID- 20022830 TI - [Clinical questions, levels of evidence and recommendations]. PMID- 20022831 TI - [Treatment with antiviral agents]. PMID- 20022832 TI - [Technical report on the 2009 influenza A (H1N1) pandemic]. AB - INTRODUCTION: Since its appearance in April 2009, the influenza A (H1N1) pandemic has been a subject of continued attention by national and international health authorities, as well as in the communication media. It has been six months since the first cases were published and the winter season has just ended in the southern hemisphere. Therefore, we now have quite extensive knowledge on the behaviour of the disease, its severity and the way it manifests itself in the child/adolescent population. The Spanish Paediatric Association commissioned its Evidence Based Medicine Working Group to prepare a technical report on the influenza pandemic. This report has been prepared following the highly structured working methodology proposed by the so-called Evidence Based Medicine (EBM). This methodology requires formulating clinical questions, carrying out a systematic review of the literature looking for research works that could answer them, the critical reading of these, evaluating their methodology quality and clinical importance and finally, establishing recommendations based on those studies considered valid and important as well as on good clinical judgement. SCOPE: The present report approaches all aspects of the influenza pandemic considered to be of interest: extent of the disease, clinical and laboratory diagnosis, physical prevention measures, vaccination and pharmacological treatment. The target population of the report are children and adolescents. Many of the considerations made may also be applied to other age groups. OBJECTIVES: The primary objective of this report is to establish a group of recommendations which may serve as a generic framework for the prevention, diagnosis and treatment of the pandemic influenza in children and adolescents. The final targets of the report are paediatricians and also general/family doctors and nurses who look after children and adolescents. PMID- 20022833 TI - Increased culture sensitivity with direct inoculation of seroma fluid in blood culture bottles. PMID- 20022834 TI - Remodelling of the pinna in myxoid degeneration of the ear. AB - Idiopathic deformation of the pinna is not widely reported in the current literature. We present a series of cases in which patients have required surgery for spontaneous thickening and deformation of the auricular cartilage, and a description of a technique for surgical correction. Four cases of idiopathic deformation of the pinna are reported. Our preferred technique of scaphoid rim incision and anterior carving of the cartilage is described, with intra-operative photographs. Each patient reported spontaneous swelling of the upper poles of the ears beginning in the second or third decade of life. In 3 cases the deformity was bilateral, although in each of these cases one side was more severely affected than the other. Histology for these cases was reported as myxoid degeneration of the ear. All of the reported patients were pleased with the aesthetic outcome of their auricular remodelling. Currently, there are no typical patient demographics for idiopathic myxoid degeneration of the ear. We have achieved good aesthetic results by hand carving the anterior aspect of the deformed cartilage via scaphoid rim incisions. PMID- 20022835 TI - Orthotic devices and gait in polio patients. AB - Polio survivors are aging and facing multiple pathologies. With age, walking becomes more difficult, partly due to locomotor deficits but also as a result of weight gain, osteoarticular degeneration, pain, cardiorespiratory problems or even post polio syndrome (PPS). These additional complications increase the risk of falls in this population where the risk of fractures is already quite high. The key joint is the knee. The muscles stabilizing this joint are often weak and patients develop compensatory gait strategies, which could be harmful to the locomotor system at medium or long term. Classically, knee recurvatum is used to lock the knee during weight bearing; however, if it exceeds 10 degrees , the knee becomes unstable and walking is unsafe. Thus, regular medical monitoring is necessary. Orthoses play an important role in the therapeutic care of polio survivors. The aim is usually to secure the knee, preventing excessive recurvatum while respecting the patient's own gait. Orthoses must be light and pressure-free if they are to be tolerated and therefore effective. Other joints present fewer problems and orthoses are rarely indicated just for them. The main issue lies in the prior evaluation of treatments' impact. Some deformities may be helpful for the patients' gait and, therefore, corrections may worsen their gait, especially if a realignment of segments is attempted. It is therefore essential to carefully pre-assess any change brought to the orthoses as well as proper indications for corrective surgery. In addition, it is essential for the patient to be monitored by a specialized team. PMID- 20022836 TI - Efficacy of genotype notification to Japanese smokers on smoking cessation--an intervention study at workplace. AB - OBJECTIVES: It is well-known that smoking causes many diseases including cancers. Informing smokers of their genotypes associated with the vulnerability to the harms of smoking may be effective measures for smoking cessation. The present study examined the effects of genotype notification of an oncogene (L-myc) genotype to smokers on their behavior to quit smoking. METHODS: Subjects were 562 employees of a bank who answered to be a smoker for a questionnaire used at annual health checkup at workplace from July to December 2002. Those enrolled on August, October, and December were allocated into the genotype notification group (intervention group), and the rest into the controls. Among 286 smokers allocated into the intervention group, 257 participants (89.9%) agreed to genotype testing. One year after the enrollment, a follow-up questionnaire survey was conducted for all smokers including controls. RESULTS: Those who stated to have quitted smoking were 22 (8.0%) among the 276 controls and 15 (5.8%) among the 257 genotype notified participants, providing that the odds ratio (OR) of cessation for the intervention was 0.64 (95% confidence interval, 0.32-1.28). No psychological problems associated with genotype notification were observed. CONCLUSION: The present study did not show positive effects of genotype notification on smoking cessation rate. To elevate the cessation rate, methods to explain and notify genotypes should be improved. PMID- 20022837 TI - Why is the incidence of childhood cancer lower in rural India? PMID- 20022838 TI - Contributions of leukocytes to tumor invasion and metastasis: the "piggy-back" hypothesis. PMID- 20022839 TI - Under-diagnosis and under-ascertainment of cases may be the reasons for low childhood cancer incidence in rural India. AB - Cancer statistics from India revealed that childhood cancer incidence is lesser in rural than urban India. This might be due to under-diagnosis or under ascertainment of cases or could even be true. With registries able to explicitly measure and appropriately streamline the ascertainment of cases to comply with acceptable standards, it is under-diagnosis that is variable and highly influenced by development of or accessibility to specialized centres in or around the registry area. This is reflected implicitly by marked variation in incidence between different populations in India: weighted age standardized rates of all childhood cancers together was the highest (108 per million) in metropolitan areas, followed by other urban (86) and rural (53) areas in that order. A childhood cancer registry focusing on pertinent data collection and specific epidemiological studies is desirable to explain the variations in incidence and outcome of childhood cancers in India. PMID- 20022854 TI - [New drugs in metastatic castration-resistant prostate cancer]. AB - Despite that greater knowledge of prostate cancer biology has led to the isolation of many new and promising targets, treatment of metastatic prostate cancer is still challenging. New agents targeting these molecules are currently under development in large randomized phase III trials, to improve overall survival and the quality of life of patients with metastatic castrate-resistant prostatic cancer (CRPC). Cytotoxic chemotherapy (docetaxel-based chemotherapy) demonstrated clinical benefit on overall survival, but could be improved. Drugs targeting directly or not the androgen receptor such as abiraterone or new specific peripheral anti-androgens (MDV3100) are very promising. Bone targeted therapies (endothelin1 receptor A inhibitor, RANK ligant, metabolic irradiation) are also very promising and are in development in large phase III trials. Antiangiogenic therapies could also be effective in CRPC. Autologous vaccin against prostatic acid phosphatase seems to prolong overall survival and other vaccin and immunotherapy strategies are in development (anti-CTLA4 antibody). A recent analogue of thalidomide, probably more efficient, lenalidomide is also in development. PMID- 20022855 TI - Indoor air studies of fungi contamination at the Department of Pulmonology and Internal Medicine in Kavala Hospital in Greece. AB - PURPOSE: The aim of the study was to assess the presence of airborne fungi and the fungal flora of the walls in the Departments of Pulmonary and Internal Medicine in Kavala Hospital (Greece). MATERIAL AND METHODS: The study was carried out at the Department of Pulmonology and Internal medicine in Kavala Hospital. Materials for the tests were: the air samples (in front of the building and the selected rooms) and swabs from the walls. The air pollution was determined using SAS SUPER 100 (Pbi International). The microbial flora from walls was assessed using the Count-Tact applicator and the plate Count-Tact (BioMerieux). Humidity and temperature were evaluated by a thermo-hygrometer. RESULTS: The following fungal pathogens isolated from air were Aspergillus, Candida albicans, Candida spp., and Penicillium species in the Department of Pulmonary. Similar pathogens in the air of Department and Internal Medicine were found. Mean number of fungi colonies isolated from air in the Department of Pulmonology was significantly (p<0.001) higher compared to the Department of Internal Medicine. No significant correlations between CFU of fungi in air and temperature in both Departments were found. CONCLUSIONS: The main fungal pathogens isolated from the air samples were Aspergillus and Candida albicans in the Departments of Pulmonary and Internal Medicine in Kavala Hospital. Fungal occurrence in the air of rooms and walls varied between the both departments of the same hospital. PMID- 20022856 TI - Serum concentration of adiponectin, leptin and resistin in obese children with non-alcoholic fatty liver disease. AB - PURPOSE: Obesity, insulin resistance and dyslipidemia are the most significant risk factors of non-alcoholic fatty liver disease (NAFLD) but the role of adipokines in patomechanism of this disease is not clear. The aim of the study was to evaluate the serum levels of leptin, adiponectin and resistin in obese children with NAFLD. MATERIAL/METHODS: The fasting serum levels of adipokines were determined in 44 consecutive obese children with suspected liver disease and in 24 lean controls. The degree of the ultrasound liver steatosis was graded according to Saverymuttu. RESULTS: The fatty liver was confirmed in 33 children by ultrasonography (16 of them also showed an increased ALT activity). The serum leptin level was significantly higher and adiponectin level was lower in the obese children with NAFLD when compared to controls. Only adiponectin correlated with homeostasis model assessment of insulin resistance (HOMA-IR). Significant negative correlations were found between the ultrasonographic grades of liver steatosis and adiponectin and resistin levels. Serum adiponectin and resistin levels were lower in children with an advanced liver steatosis (grade 3, n=10) compared to patients with a mild steatosis (grade 1-2, n=23). The ability of serum adiponectin and resistin to differentiate children with an advanced liver steatosis from those with mild steatosis was significant. CONCLUSIONS: These data suggest a role of both adiponectin and resistin in the pathogenesis of NAFLD in obese children and confirm the association between adiponectin and insulin resistance. Adiponectin and resistin may be suitable serum markers in predicting an advanced liver steatosis in children with NAFLD. PMID- 20022857 TI - Norovirus and Rotavirus--two major causative agents of sporadic viral gastroenteritis in hospitalized Polish children. AB - PURPOSE: The aim of the study was to establish the main etiologic agents of acute viral gastroenteritis and to asses the severity of illness associated with the different viral agents of gastroenteritis in children hospitalized during spring/summer season 2008. MATERIAL AND METHODS: 181 stool specimens were collected from children under 5 years of age, hospitalized with acute viral diarrhea from April to September 2008. Commercial enzyme immunoassay kits were used to detect noroviruses. The immunochromatographic tests for combined detection of rotaviruses and adenoviruses were performed routinely in our department in all in-patients suspected for viral gastroenteritis. RESULTS: A viral etiologic agents were detected in 108 of 181 (59.7%) samples tested. Dual viral pathogens (rotavirus and norovirus) were found in 3 of 181 (1.7%) samples. Rotavirus was the most common viral pathogen found in the study group (86/181; 47.5%), followed by NV (19/181; 10.5%) and adenoviruses (3/181; 1.7%). Approximately, 60% of acute gastroenteritis episodes occurring in children less than 5 years of age were accounted for by infection due to rotavirus and/or norovirus. Norovirus cases were clinically indistinguishable from those of rotavirus origin in children aged less than 2 years whereas they were slightly milder in older group of patients. CONCLUSIONS: Rotavirus infections are leading cause of children's hospitalization in spring months whereas Norovirus infections during spring/summer time. There is a great need to apply molecular diagnostic tools to determine the actual and monitoring the changing etiology of acute enteritis in Polish population. PMID- 20022858 TI - Surgery within multimodal therapy concepts for esophageal squamous cell carcinoma (ESCC): the MRI approach and review of the literature. AB - BACKGROUND: Radical esophagectomy with lymphadenectomy remains the only curative therapy for patients with resectable esophageal squamous cell cancer (ESCC), however, combined treatment modalities may improve survival. Based upon more than 1300 consecutive esophageal resections, we present our current multidisciplinary ESCC approach with analysis in the context of recently published RCTs. METHODS: Subject to tumor staging, patients with resectable ESCC receive either a neoadjuvant radiochemotherapy (uT3N+) or are referred to primary surgery (uT1/2N0). By Medline searches (1997-2009), all published RCTs containing multimodal ESCC therapy concepts were identified and a systematic review was generated. RESULTS: From July 2007 to June 2009, 62 patients with ESCC were treated in our department (40 multimodal treatment concept, 21 primary surgery, 1 definite radiochemotherapy). The R0 resection rate was 78%, in hospital mortality 4.8%. 60% of patients showed a good response to neoadjuvant treatment. 18-month follow-up data revealed absence of tumor recurrence in 7 patients (18%). Our approach is aligned to the current published literature including 12 studies in this review. In line with our institutional experience, neodjuvant radiochemotherapy tends to improve overall survival and increases the likelihood of R0 resection. However, postoperative morbidity and mortality rates are increased. Adjuvant treatment failed to demonstrate any improvement in prognosis. For palliation, concurrent radiochemotherapy is the treatment of choice. CONCLUSION: The MRI approach can be aligned to the most recent published data. Surgical resection remains the principle treatment for patients with resectable ESCC. Although multimodal therapy concepts tend to improve survival rates, postoperative morbidity and mortality rates are increased. PMID- 20022859 TI - Anthropometric parameters of growth and nutritional status in children aged 6 to 7 years in R. Macedonia. AB - PURPOSE: Evaluation of age- and sex-specific differences of anthropometric parameters that were used as indicators of growth and nutritional status in children aged 6 and 7 years in R. Macedonia and their comparison with the WHO and NCHS reference values. MATERIAL AND METHODS: The study included 580 children aged 6 to 7 years from different regions of R. Macedonia. Eighteen anthropometric parameters were measured using standard equipment and measurement technique according to the International Biological Programme. The following nutritional indicators were calculated: BMI, weight-for-height, weight-for-age and height-for age. RESULTS: The majority of anthropometric parameters have shown significant age- and sex-specific differences in favour of male subjects, with exception of skin-folds that were apparently higher in female subjects. Values at the 50th percentile in our male subjects for the index weight-for-age were 23 kg and 26.95 kg, respectively, whereas in the female subjects they were 22 kg and 25 kg, respectively. The index height-for-age corresponding to the 50th percentile showed higher values in our male subjects (119.5 cm and 125.55 cm) than in our female subjects (118 cm and 124.5 cm, respectively). 50th percentile in our male subjects for BMI was 16.48 kg/m2 and 16.75 kg/m2, whereas in our female subjects it was 15.98 kg/m2 and 16.25 kg/m2, respectively. CONCLUSIONS: The found values are recommended to be applied for evaluation of deviations in the growth and nutritional status in children aged 6 and 7 years from Macedonian nationality. PMID- 20022860 TI - The GOX/CAT system: a novel enzymatic method to independently control hydrogen peroxide and hypoxia in cell culture. AB - The increasing demand in studying cellular functions in cultured cells under various levels of oxygen and hydrogen peroxide (H2O2) is only partly fulfilled by conventional approaches such as hypoxia chambers, bolus additions of H2O2 or redox-cycling drugs. This article describes the recently developed enzymatic GOX/CAT system consisting of glucose oxidase (GOX) and catalase (CAT) that allows the independent control and maintenance of both H2O2 and hypoxia in cell culture. In contrast to hypoxia chambers, the GOX/CAT system more rapidly induces hypoxia within minutes at a defined rate. The degree of hypoxia is dependent on the GOX activity and the diffusion distance of oxygen from the medium surface to the adherent cells. In contrast, H2O2 levels are solely controlled by the ratio of GOX and CAT activities. They can be adjusted at non-toxic or toxic dosages over 24 hours. Thus, the GOX/CAT system mimics a non-phosphorylating respiratory chain and allows to adjust H2O2 levels under hypoxic conditions truly simulating H2O2 release e.g. by inflammatory cells or intracellular sources. GOX/CAT can be employed to address many questions ranging from redox signaling to ischemia/reperfusion studies in transplantation medicine. Factors such as HIF1 alpha that respond both to hypoxia and H2O2 are an especially attractive target for the novel methodology. Several applications are discussed in detail to demonstrate the technical requirements and potentials. In addition, simplified protocols are presented for cell or molecular biology labs without dedicated biophysical equipment. PMID- 20022861 TI - Effect of MPEP on rat's behavioral activity in experimental episodes of hypoxia. AB - PURPOSE: The influence of the selective antagonism of metabotropic glutamate receptor subtype 5 (mGluR5) by MPEP (2-methyl-6-(phenylethynyl)-pyridine) on some behaviors was tested in control groups of rats and in rats exposed to short-term hypoxia once or to repeated episodes of hypoxia. MATERIAL AND METHODS: We used the following methods: the open field test, the passive avoidance test and the object recognition test. Experimental hypoxia was produced by placing rats in a glass chamber flushed with a mixture of 2% O2 in N2. RESULTS: MPEP applied intravenously (IV) at the dose of 1 mg kg-1 significantly enhanced locomotor and exploratory activity, impaired acquisition, but improved consolidation and retrieval in the passive avoidance situation and did not alter rats' activity in the object recognition test. The single short-term hypoxia significantly inhibited motility of rats and profoundly impaired acquisition, consolidation and retrieval processes, but the positive effect of MPEP on retrieval was preserved. Hypoxia also did not influence the activity of rats in the object recognition object. The repeated episodes of short-term hypoxia were induced for five consecutive days and it also inhibited motility of rats, but did not change consolidation and retrieval processes. The episodes of hypoxia significantly diminished the beneficial effect of MPEP on consolidation and retrieval, and also the enhancement of locomotor and exploratory activity. MPEP, used in rats subjected to the single or the repeated episodes of short-term hypoxia, did not change recognition memory. CONCLUSION: MPEP used before the single episode of hypoxia only, had beneficial effect on retrieval. PMID- 20022862 TI - Influence of stress related to war on biological and morphological characteristics of breast cancer in a defined population. AB - PURPOSE: To assess differences between patients with breast cancer before, during and after the 1991-1995 war in Croatia. MATERIAL AND METHODS: We analyzed 660 patients of Pozesko-Slavonska County, during the three periods. Relative predictive values of patient's characteristics and stage of tumor were assessed using the X2-test, and survival with Kaplan-Meier analysis. RESULTS: Tumors were significantly more often of higher stages (IIA and IIB) and with axillary lymph node metastases (N1) during the war, than in pre-war and post-war period.Breast cancer was significantly more frequent in patients who previously experienced death in the family (35,3%). The Kaplan-Meier analysis showed correlation between survival, T and N stages of tumor and clinical stage of tumor. CONCLUSION: The war aggression towards Croatia with its impact on our patients, contributed to modification of characteristics of breast cancer in the analyzed period. PMID- 20022863 TI - Remobilization of acorn nitrogen for seedling growth in holm oak (Quercus ilex), cultivated with contrasting nutrient availability. AB - The relative contribution of nitrogen (N) reserves from seeds or uptake by the roots to the growth and N content of young seedlings has received little attention. In this study, we investigated the contribution of N from the acorn or uptake by the roots to the N content of holm oak (Quercus ilex L.) seedlings and determined if remobilization of acorn N was affected by nutrient availability in the growing media. Q. ilex seedlings were cultivated for 3 months, until the end of the second shoot flush of growth, with three N fertilization rates: 8.6 mM N, 1.4 mM N or no fertilization. Fertilizer N was enriched in (15)N. Between 62 and 75% of the N contained in high and low fertilized seedlings, respectively, at the end of the second flush of growth was derived from the acorn. However, the dependence on acorn N was greater during the early root growth and first shoot flush of growth and decreased during the second shoot flush of growth, with root uptake contributing 32-54% of plant new N in this latter developmental stage in high and low fertilized plants, respectively. Fertilization rate did not affect the amount of N taken up during the earliest developmental stages, but it increased it during the second shoot flush of growth. Fertilization increased the mass of the shoot segment formed during the second shoot flush of growth and reduced the root mass, with no effect on whole plant growth. Remobilization of acorn N was faster in unfertilized plants than in fertilized plants. It is concluded that the holm oak seedlings depend greatly upon acorn N until the end of the second shoot flush of growth, that significant root N uptake starts at the beginning of the second shoot flush of growth and that acorn N remobilization is a plastic process that is accelerated under extremely low substratum nutrient content. PMID- 20022864 TI - Bud dormancy release in elm (Ulmus spp.) clones--a case study of photoperiod and temperature responses. AB - Dormancy release as influenced by duration of outdoor winter chilling in Florence (Italy) was studied under different photoperiodic and temperature treatments in collected twigs of two European (Ulmus glabra Huds. and Ulmus minor Mill.) and four Asian (Ulmus pumila L., Ulmus parvifolia Jacq., Ulmus macrocarpa Hance and Ulmus villosa Brandis) elm clones. Photoperiod had no effect on dormancy release, and there was no evidence that photoperiod affected bud burst during quiescence in the studied elm clones. Thermal time (day degrees >0 degrees C) to bud burst decreased in all the clones with increasing outdoor chilling. Although all the clones exhibited a rather weak dormancy, they significantly differed from each other. Dormancy was released earlier in the Asian than in the European clones, and the clones could be ranked from the U. pumila clone (very weak and short dormancy) to the U. minor clone (relatively stronger and longer dormancy), the other clones being intermediate. In all the clones except U. minor, the observed decrement in thermal time to bud burst was efficiently explained as an inverse exponential function of the number of chill days < or =5 degrees C received outdoor in autumn and winter. Endodormancy, as measured by the single-node cuttings test, was weak and short in all the clones. The latter result suggests that correlative inhibitions were largely responsible for preventing bud burst during winter in these elm clones. PMID- 20022865 TI - The influence of micropropagation on growth and coppicing ability of Eucalyptus polybractea. AB - A micropropagation protocol was recently developed for Eucalyptus polybractea R.T. Baker, a commercially important eucalypt grown in short-rotation coppice cultivation and harvested for its foliar 1,8-cineole oil. Micropropagation of elite E. polybractea trees has resulted in selection gains for foliar oil traits, but decreased above-ground biomass accumulation has been observed in clones compared to related half-sibling families. This study aims to use a greenhouse study to investigate if micropropagation induces somaclonal variation that can account for the reduction in above-ground biomass in E. polybractea clones. Secondly, the study aims to compare the coppicing ability of micropropagated clones with related half-sibling seedlings using de-topped plantation-grown saplings. The results of the greenhouse study suggest that micropropagation of E. polybractea induces somaclonal variation that manifests in more mature leaf morphologies such as increased foliar oil concentrations and lower specific leaf area (SLA), attributable to an isobilateral arrangement of increased palisade mesophyll layers. Lower SLA, rather than differences in root allocation, is likely to be a key contributor to the lower relative growth rates observed in early sapling growth of micropropagated clones. In the field study, all micropropagated and seedling-derived E. polybractea saplings coppiced vigorously in the 12 months after de-topping. The coppice growth was so vigorous in the 12 months after de-topping that total above-ground biomass equalled that of the 27 month-old saplings, irrespective of propagation source. The morphological distinction between leaves of micropropagated and seed-derived plants was no longer evident in the coppice regrowth. The results presented here suggest that the micropropagated leaf morphology and the resultant growth reduction is transient and micropropagated plants coppice just as vigorously as seed-derived plants. Therefore, micropropagation is unlikely to detrimentally influence above ground biomass accumulation beyond the first harvest rotation. PMID- 20022866 TI - cDNA-AFLP analysis of gene expression in hickory (Carya cathayensis) during graft process. AB - The graft technique is a valid method for propagating plants. A better elucidation of the graft mechanism is helpful in improving the production efficiency and fruit quality in hickory. In this study, cDNA-amplified fragment length polymorphism analysis was used to examine the gene expression in hickory at four time points (at 0, 3, 7 and 14 days) during the graft process. Forty-nine unique genes involved in the graft mechanism were obtained. The expression patterns of these genes were confirmed by real-time reverse transcription polymerase chain reaction analysis based on 12 selected genes representing different patterns. The 49 genes composed 19 genes of known function, nine genes of unknown function and 21 novel genes. These 19 genes of known functions were involved in the indole-3-acetic acid transport protein, cell cycle, signal transduction, water metabolism, nuclear metabolism, amino acid metabolism, protein metabolism, carbon metabolism and secretion of substances, suggesting that Carya cathayensis Sarg. undergoes a complex metabolism process during the grafting. PMID- 20022867 TI - Inhibitor studies of leaf lamina hydraulic conductance in trembling aspen (Populus tremuloides Michx.) leaves. AB - The present study investigated leaf water transport properties in trembling aspen (Populus tremuloides) leaves. Leaf lamina hydraulic conductance (K(lam)) and stomatal conductance (g(s)) were drastically suppressed by NaF (a general metabolic inhibitor). In leaves treated with 0.2 mM HgCl(2) (an aquaporin blocker), K(lam) declined by 22% when the leaves were sampled in June but the decline was not significant when the leaves were sampled in August. The leaves sampled in June that transpired 30 mM beta-mercaptoethanol following mercury application showed similar K(lam) as those in control leaves transpiring distilled water. When leaves were pressure-infiltrated with 0.1 mM HgCl(2), K(lam) significantly declined by 25%. Atrazine (a photosystem II inhibitor) drastically reduced leaf net CO(2) uptake by the leaves from seedlings and mature trees but did not have any effect on K(lam) regardless of the irradiance at the leaf level during the K(lam) measurements. When PTS(3) (trisodium 3-hydroxy 5,8,10-pyrenetrisulphonate) apoplastic tracer was pressure-infiltrated inside the leaves, its concentration in the leaf exudates did not change from ambient light to high irradiance treatment and declined in the presence of HgCl(2) in the treatment solution. Trembling aspen K(lam) appears to be linked to leaf metabolism and is uncoupled from the short-term variations in photosynthesis. Aquaporin-mediated water transport does not appear to constitute the dominant pathway for the pressure-driven water flow in the leaves of trembling aspen trees. PMID- 20022868 TI - Will simultaneous measurement of E/e' index facilitate the non-invasive assessment of left ventricular filling pressure in patients with non-valvular atrial fibrillation? AB - AIMS: The ratio of early transmitral flow velocity (E) to early diastolic mitral annular velocity (e') was applied as a non-invasive index to assess left ventricular filling pressure (LVFP). However, the reliability of E/e' was undermined in patients with atrial fibrillation (AF). Recently, a novel method entitled 'dual Doppler' was established, which allows simultaneous recording and display of E and e'. Our study investigates whether the dual-Doppler method improves the reliability of E/e' in AF patients. METHODS AND RESULTS: Forty-nine patients with non-valvular AF underwent conventional echocardiography, dual Doppler echocardiography, and cardiac catheterization within 4 h. Of 22 patients (45%c) with increased LVFP (>15 mmHg), higher E/e' measured by both conventional tissue Doppler imaging (TDI) and dual-Doppler method was observed. Conventional echocardiographic variables were correlated with LVFP (E/e'(sep), r = 0.404, E/e'(lat), r = 0.487), but E/e' measured synchronously in the dual-Doppler mode yielded a better correlation (E/e'(synchronous sep), r = 0.754, E/e'(synchronous lat), r = 0.765). The intra- and interobserver variability of the dual-Doppler method was significantly lower than the conventional TDI method. CONCLUSION: Good correlations were found between E/e'' and LVFP in patients with AF, particularly in the dual-Doppler mode. E/e' measured by the dual-Doppler method can therefore be applied to assess diastolic dysfunction in AF patients. PMID- 20022869 TI - Pitfalls of anatomical aortic valve area measurements using two-dimensional transoesophageal echocardiography and the potential of three-dimensional transoesophageal echocardiography. AB - AIMS: The aims of this study were to (i) investigate aortic annulus dynamics using two-dimensional (2D) speckle tracking echocardiography, (ii) determine optimal 2D short-axis view for the calculation of planimetric aortic valve area (AVA), and (iii) compare 2D planimetric AVA extracted from volumetric three dimensional data sets using real-time 3DTEE (three-dimensional transoesophageal echocardiography) with standard 2DTEE planimetry. METHODS AND RESULTS: We studied 60 patients with aortic stenosis (AS) and 10 control subjects. AVA was calculated by standard 2DTEE planimetry method, volumetric 3DTEE method, and continuity equation (CE) from transthoracic echocardiography. In addition, aortic annular motion was studied using 2D speckle tracking. Aortic annulus moves cranially during early systole and subsequently moves caudally during the remainder of systole and isovolumic relaxation. Annulus again moved in the cranial direction during diastole in both groups. Although AVA correlated well between 2DTEE and 3DTEE methods (r = 0.95), 2DTEE showed a significantly larger AVA compared with 3DTEE method (1.26 +/- 0.39 vs. 1.10 +/- 0.39 cm(2), P < 0.001). In patients in whom aortic cusps were visible in 2DTEE short-axis images during systole only (n = 45), AVA using 2DTEE was still larger than that measured with 3DTEE. However, the bias in AVA was significantly lower compared with the remaining 15 patients ( 0.13 +/- 0.11 vs. -0.26 +/- 0.12 cm(2), P < 0.005). Although both methods showed moderate correlation with AVA by CE (r = 0.78, 0.75), mean differences were significantly smaller by 3DTEE than 2DTEE (-0.01 +/- 0.25 vs. -0.17 +/- 0.27 cm(2), P < 0.001). CONCLUSION: Aortic annular motion affects the calculation of AVA using 2DTEE. Three-dimensional transoesophageal echocardiography has a potential for more accurate determination of anatomical AVA. PMID- 20022870 TI - Diagnosis of the prosthetic heart valve pannus formation with real-time three dimensional transoesophageal echocardiography. AB - Prosthetic heart valve obstruction (PHVO) caused by pannus formation is an uncommon but serious complication. Although two-dimensional transesophageal echocardiography (2D-TEE) is the method of choice in the evaluation of PHVO, visualization of pannus is almost impossible with 2D-TEE. While demonstrating the precise aetiology of PHVO is essential for guiding the therapy, either thrombolysis for valve thrombosis or surgery for pannus formation, more sophisticated imaging techniques are needed in patients with suspected pannus formation. We present real-time 3D-TEE imaging in a patient with mechanical mitral PHVO, clearly demonstrating pannus overgrowth. PMID- 20022871 TI - Isolated mitral valve prolapse is an independent predictor of aortic root size in a general population. AB - AIMS: Mitral valve prolapse (MVP) is associated with aortic root (AoR) enlargement in patients with inherited connective tissue disorders. This report evaluated whether MVP is related to AoR dimension in a large population with otherwise normal echocardiographic parameters. METHODS AND RESULTS: We retrospectively analysed echocardiograms performed by a single echocardiographer between 2001 and 2007 for various clinical indications. Six hundred and twenty seven subjects with isolated MVP were found and then matched by sex, age, and body mass index to 627 individuals without MVP. The whole sample included 454 men and 800 women with an average age of 37.9 +/- 0.3 years and a body mass index of 23.7 +/- 0.1 kg/m(2). MVP subjects had a higher AoR diameter (30.4 +/- 0.1 vs. 29.5 +/- 0.1 cm; P < 0.0001) compared with controls. Furthermore, multivariate analyses demonstrated an independent association between MVP and AoR size (P < 0.0001) in a model that included age, gender, body mass index, body surface area, blood pressure levels, and left ventricular mass index as confounding variables. CONCLUSION: Isolated MVP is an independent predictor of greater AoR size in a large population with otherwise normal echocardiographic parameters. PMID- 20022872 TI - Influence of bone marrow stem cells on left ventricle perfusion and ejection fraction in patients with acute myocardial infarction of anterior wall: randomized clinical trial: Impact of bone marrow stem cell intracoronary infusion on improvement of microcirculation. AB - AIMS: Randomized trial to assess change in left ventricle ejection fraction (LVEF) and myocardial perfusion in patients with acute myocardial infarction (AMI) of anterior wall treated with bone marrow stem cells (BMSCs), compared with control group-from baseline in the acute phase up to 12 months of follow-up. METHODS AND RESULTS: Forty-five patients were randomized 2:1 to BMSC group (n= 31) or to control group (n = 14). Bone marrow stem cells were administered into infarct-related artery (IRA) at 4-6 day after primary PCI. Groups were followed up with Tc-99m-MIBI SPECT, radionuclide ventriculography (EF-RNV), echocardiography (ECHO), and spiroergometric stress test. Coronary angiography was repeated after 6 months. EF-RNV did not differ significantly in both groups, but trend towards increase in EF at 6 months and its maintenance after 12 months was noticed in the BMSC group. At rest study, perfusion index (PI) of region supplied with blood by IRA distal to its previous occlusion (PI-IRA) improved significantly in the BMSC group at 6 months: PI-IRA at 4-6 days vs. PI-IRA at 6 months (3.00 +/- 0.97 vs. 2.65 +/- 0.64; P = 0.017). At 12 months, PI-IRA at rest was 2.66 +/- 0.55; P = 0.07. The difference between BMSC and control groups at rest study in PI-IRA was not observed. At dipyridamole study (PI-dip), perfusion in the BMSC group was better compared with controls at 6 months (2.26 +/- 0.44 vs. 2.47 +/- 0.40; P = 0.033) and at 12 months (2.34 +/- 0.55 vs. 2.52 +/- 0.42; P = 0.014), also for region supplied with blood by IRA (PI-IRA-dip; at 6 months 2.63 +/- 0.77 vs. 3.06 +/- 0.46; P = 0.021 and at 12 months 2.71 +/- 0.63 vs. 3.15 +/- 0.51; P = 0.001). Results of LVEF, LVEDV, LVESV in ECHO and results of spiroergometric stress test did not differ significantly between groups. Major adverse cardiac events occurred more often in the control group (P = 0.027). CONCLUSION: In our study, BMSC intracoronary transplantation in patients with anterior AMI did not result in increase in EF. Slight improvement of myocardial perfusion was noticed in the BMSC group. This finding may indicate better microcirculation enhanced by BMSCs, but small number of patients allow for hypothesis rather than final statement. PMID- 20022873 TI - Rosiglitazone reduces ischaemia-reperfusion injury in patients with the metabolic syndrome. PMID- 20022874 TI - Intracardiac echocardiography identifies the substrate of left ventricular papillary muscle ectopy. PMID- 20022875 TI - Myocardial perfusion imaging does not adequately assess the risk of coronary artery disease in patients with atrial fibrillation. AB - AIMS: The role of coronary artery disease (CAD) in atrial fibrillation (AF) is poorly investigated. This study investigated the value of myocardial perfusion single-photon emission computed tomography (SPECT) in the assessment of risk of CAD in patients with a history of AF. METHODS AND RESULTS: Out of consecutive patients without previous coronary angiogram or history of CAD referred for SPECT, patients with a history of AF (n = 129) were compared with age- and gender matched controls (n = 124). Primary endpoint was positive SPECT, i.e. unambiguous signs of ischaemia. There was no significant difference with regard to positive SPECT outcome between AF patients and controls (14 patients, 11% vs. 21 patients, 17%; P = 0.16). Coronary angiography (CAG) performed after SPECT demonstrated a higher yield of positive SPECT regarding significant CAD in control patients (10 out of 15 patients, 67%) than in AF patients (2 out of 13 patients, 15%; P = 0.006). CONCLUSION: Positive SPECT outcome was similar in patients with AF and in controls. Nevertheless, in AF patients a positive SPECT outcome was less often related to significant CAD in those patients who subsequently underwent CAG. These results emphasize the need for new non-invasive techniques to adequately assess the risk of significant CAD in AF patients. PMID- 20022876 TI - Increase in organization index predicts atrial fibrillation termination with flecainide post-ablation: spectral analysis of intracardiac electrograms. AB - AIMS: The mechanism of the action of flecainide in the termination of human atrial fibrillation (AF) is not fully understood. We studied the acute effects of flecainide on AF electrograms in the time and frequency domain to identify factors associated with AF termination. METHODS AND RESULTS: Patients who were still in AF at the end of catheter ablation for AF were given intravenous flecainide. Dominant frequency (DF) and organization index (OI) were obtained by fast Fourier transform of electrograms from the coronary sinus catheter over 10 s in AF, before and after flecainide infusion. Mean AF cycle length (CL) was also calculated. Twenty-six patients were studied (16 paroxysmal AF and 10 persistent AF). Seven converted to sinus rhythm (SR) with flecainide. In all patients, mean CL increased from 211 +/- 44 to 321 +/- 85 ms (P < 0.001). Mean DF decreased from 5.2 +/- 1.03 to 3.6 +/- 1.04 Hz (P < 0.001). Mean OI was 0.33 +/- 0.13 before and 0.32 +/- 0.11 after flecainide (P = 0.90). Comparing patients who converted to SR with those who did not, OI post-flecainide was 0.41 +/- 0.12 vs. 0.29 +/- 0.10 (P = 0.013), and the relative change in OI was 29 +/- 33 vs. -3.9 +/- 27% (P = 0.016), respectively. No significant difference was noted in the change in CL and DF in the two groups. CONCLUSION: Increase in OI, independent of changes to CL and DF, appears critical to AF termination with flecainide. Increase in OI holds promise as a sensitive predictor of AF termination. PMID- 20022877 TI - Free floating right atrial thrombus in two asymptomatic patients after electrophysiological study: role of routine echocardiography after ablation. AB - This report presents two patients with free floating right atrial and atrioventricular thrombi after electrophysiology study which were treated with thrombolysis in the first patient and surgical operation in the second patient without any complication. PMID- 20022878 TI - An original defibrillation lead implantation to avoid tricuspid prosthesis damage. AB - We report the case of a 67-year-old woman suffering from Ebstein's disease, who underwent three cardiac operations for bypass, tricuspid prosthesis and pacemaker implantation, and who needed an implantable cardioverter defibrillator for recurrent syncopes related to ventricular tachycardia. Because of the tricuspid prosthesis we chose to implant the defibrillation lead in the inferior vena cava. We collected satisfactory pacing and sensing data and performed a successful defibrillation test during the procedure. This configuration appears to be a safe alternative to conventional implantation in the coronary sinus, as already described in the literature for a few cases. PMID- 20022879 TI - Gold-coated pacemaker implantation after allergic reactions to pacemaker compounds. AB - An 86-year-old man underwent pacemaker implantation for symptomatic atrio ventricular block grade 2 Mobitz II. The patient suffered repeated admissions for iterative sterile wound necrosis, leading to two generator re-implantations. No bacterial infection was detected in the microbiological screening tests. The skin patch testing to titanium was negative. Nevertheless, we decided to remove the pacemaker system and to implant a gold-plated generator with polyurethane leads. Since then, there has been no recurrence of wound complications. Gold-plated generator and polyurethane leads are effective in treating allergic reactions to pacemaker system components in selected cases. Negative skin patch testing to titanium does not exclude allergic reaction to this pacemaker component. PMID- 20022880 TI - Three-dimensional cardiac rotational angiography: effective radiation dose and image quality implications. AB - AIMS: Three-dimensional rotational angiography (3DRA) is a promising new online tool for 3D imaging during cardiac ablation procedures. No precise data exist concerning its associated radiation dose. The current study evaluated the effective dose (ED) of cardiac rotational angiography and its relation to patient properties, imaging system input settings, and quality of reconstructed 3D images. METHODS AND RESULTS: We performed Monte Carlo simulation-based radiation dose calculations in 42 patients referred for ablation of cardiac arrhythmias. Detailed tube setting information from the 3DRA system (Siemens Axiom Artis dBC with Syngo DynaCT Cardiac software) was used to provide an accurate input for dose calculations in all 248 frames used during image acquisition. Our calculations yielded an overall mean ED of 6.6 +/- 1.8 mSv (based on ICRP 103 weighing factors). Manual collimation of the radiation beam can reduce ED by more than 20%. Image quality did not significantly relate to patient body mass index (BMI), dose per frame setting, or dose-area product (DAP), but was rather explained by contrast filling, cardiac motion reduction, and absence of image reconstruction artefacts. In the system evaluated, DAP values are nearly independent from BMI (R(2) = 0.30), due to its technical specifications. Therefore, patient BMI showed an unexpected strong inverse relation to ED. CONCLUSION: Three-dimensional rotational angiography can be performed with acceptable patient radiation dose, comparable to cardiac CT. With the 3DRA system studied (Siemens Axiom), slender patients may currently receive unnecessarily high radiation doses when compared with obese patients, so that further dose reduction seems feasible for many patients. Adequate collimation is imperative to limit patient exposure. PMID- 20022881 TI - Cervico-mediastinal goiter: is telescopic exploration of the mediastinum (video mediastinoscopy) useful? AB - Surgeons are aware that most mediastinal goiters can be excised through a Kocher transverse collar incision, but in rare circumstances a partial-complete median sternotomy or a thoracotomy are mandatory. During an operation to remove a large cervico-mediastinal goiter (CMG) a profound, not massive, bleeding in the anterior mediastinum developed. Bleeding was unsuccessfully treated with packing. Instead, to perform an urgent sternotomy we used telescopic imaging to identify the source of hemorrhage, and a metallic clip was used to stop the bleeding. Since then we have prospectively used the telescope in the case of large CMG causing compression of an adjacent structure. This report is a preliminary communication demonstrating the technique. Telescopic exploration of the mediastinum was performed in seven patients. The goiters were located in the middle mediastinum in five patients and in the anterior and middle mediastinum in one, respectively. The use of a telescope can help the surgeon during the removal of a large mediastinal goiter. It facilitates a) the visualization of the intrathoracic tributaries reducing the risk of hemorrhage, b) the research of ectopic thyroid gland, and finally c) minimizes the risk of complications of a median sternotomy. PMID- 20022882 TI - Surgical resection of solitary cardiophrenic lymph node metastasis by video assisted thoracic surgery after complete resection of hepatocellular carcinoma. AB - This report presents the case of a patient that developed a postoperative metastatic tumor in the cardiophrenic lymph node as a rare pattern of distant lymph node metastases of hepatocellular carcinoma (HCC) after a complete resection. This is the case of surgically and pathologically proved cardiophrenic lymph node metastasis of HCC using video-assisted thoracic surgery. General thoracic surgeons should therefore be aware of the possibility of this rare form of extrahepatic recurrence when a growing nodule is found in the pericardial fat pad during the follow-up of a malignancy in the liver. PMID- 20022883 TI - Separation anxiety: detachment from the extracellular matrix induces metabolic changes that can stimulate tumorigenesis. AB - One of the earliest stages of tumor progression involves the ability of cells to survive and proliferate when not attached to the extracellular matrix (ECM). New research using a physiologically relevant breast cancer model reveals how separation from the ECM stimulates metabolic changes characteristic of developing tumors. PMID- 20022884 TI - STING-ing the antiviral pathway. AB - The cytosolic DNA sensing pathway has remained poorly defined thus far. A recent study by Ishikawa et al. demonstrates that STING is essential for DNA-mediated type I IFN production and host defence against DNA pathogens. PMID- 20022885 TI - Interaction between mutations in the slide helix of Kir6.2 associated with neonatal diabetes and neurological symptoms. AB - ATP-sensitive potassium (K(ATP)) channels regulate insulin secretion from pancreatic beta-cells. Gain-of-function mutations in the genes encoding the Kir6.2 and SUR1 subunits of this channel cause neonatal diabetes. We report two novel mutations on the same haplotype (cis), F60Y and V64L, in the slide helix of Kir6.2 in a patient with neonatal diabetes, developmental delay and epilepsy. Functional analysis revealed the F60Y mutation increases the intrinsic channel open probability (Po(0)), thereby indirectly producing a marked decrease in channel inhibition by ATP and an increase in whole-cell K(ATP) currents. When expressed alone, the V64L mutation caused a small reduction in apparent ATP inhibition, by enhancing the ability of MgATP to stimulate channel activity. The V64L mutation also ameliorated the deleterious effects on the F60Y mutation when it was expressed on the same (but not a different) subunit. These data indicate that F60Y is the pathogenic mutation and reveal that interactions between slide helix residues can influence K(ATP) channel gating. PMID- 20022887 TI - Ciliary neurotrophic factor-induced sprouting preserves motor function in a mouse model of mild spinal muscular atrophy. AB - Proximal spinal muscular atrophy (SMA) is caused by homozygous loss or mutation of the SMN1 gene on human chromosome 5. Depending on the levels of SMN protein produced from a second SMN gene (SMN2), different forms of the disease are distinguished. In patients with milder forms of the disease, type III or type IV SMA that normally reach adulthood, enlargement of motor units is regularly observed. However, the underlying mechanisms are not understood. Smn(+/-) mice, a mouse model of type III/IV SMA, reveal progressive loss of motor neurons and denervation of motor endplates starting at 4 weeks of age. Loss of spinal motor neurons between 1 month and 12 months reaches 40%, whereas muscle strength is not reduced. In these animals, amplitude of single motor unit action potentials in the gastrocnemic muscle is increased more than 2-fold. Confocal analysis reveals pronounced sprouting of innervating motor axons. As ciliary neurotrophic factor (CNTF) is highly expressed in Schwann cells, we investigated its role for a compensatory sprouting response and maintenance of muscle strength in this mouse model. Genetic ablation of CNTF results in reduced sprouting and decline of muscle strength in Smn(+/-) mice. These findings indicate that CNTF is necessary for a sprouting response and thus enhances the size of motor units in skeletal muscles of Smn(+/-) mice. This compensatory mechanism could guide the way to new therapies for this motor neuron disease. PMID- 20022886 TI - FANCC suppresses short telomere-initiated telomere sister chromatid exchange. AB - Telomere shortening has been linked to rare human disorders that present with bone marrow failure including Fanconi anemia (FA). FANCC is one of the most commonly mutated FA genes in FA patients and the FANCC subtype tends to have a relatively early onset of bone marrow failure and hematologic malignancies. Here, we studied the role of Fancc in telomere length regulation in mice. Deletion of Fancc (Fancc(-/-)) did not affect telomerase activity, telomere length or telomeric end-capping in a mouse strain possessing intrinsically long telomeres. However, ablation of Fancc did exacerbate telomere attrition when murine bone marrow cells experienced high cell turnover after serial transplantation. When Fancc(-/-) mice were crossed into a telomerase reverse transcriptase heterozygous or null background (Tert(+/-) or Tert(-/-)) with short telomeres, Fancc deficiency led to an increase in the incidence of telomere sister chromatid exchange. In contrast, these phenotypes were not observed in Tert mutant mice with long telomeres. Our data indicate that Fancc deficiency accelerates telomere shortening during high turnover of hematopoietic cells and promotes telomere recombination initiated by short telomeres. PMID- 20022888 TI - Permuted tRNA genes in the nuclear and nucleomorph genomes of photosynthetic eukaryotes. AB - Transfer RNA (tRNA) is a central genetic element in the decoding of genome information for all of Earth's life forms. Nevertheless, there are a great number of missing tRNAs that have been left without examination, especially in microbial genomes. Two tRNA gene families remarkable in their structure and expression mechanism have been reported: split and permuted tRNAs. Split tRNAs in archaea are encoded on the genome as two or three fragmented genes and then processed into single tRNA molecules. Permuted tRNAs are organized with the 5' and 3' halves of the gene positioned in reverse on the genome and hitherto have been found only in one tiny red alga. Here we reveal a wide-ranging distribution of permuted tRNA genes in the genomes of photosynthetic eukaryotes. This includes in the smallest eukaryotic genome known to date, the nucleomorph genome of the chlorarachniophyte alga Bigelowiella natans. Comparison between cDNA and genomic DNA sequences of two nucleomorph-encoded tRNA(Ser) genes confirms that precursors are circularized and processed into mature tRNA molecules in vivo. In the tRNA(Ser)(AGA), adenine at the wobble position of the codon is likely modified to inosine to expand capacity of the codon recognition. We also show the presence of permuted tRNAs in the ultrasmall free-living green algae Ostreococcus and Micromonas, which are closely related to the B. natans nucleomorph. Conserved intron/leader sequence structures in the intron-containing and permuted tRNAs suggest the ancient origin of the splicing machinery in the common ancestor of eukaryotes and archaea. Meanwhile, a wide but patchy distribution of permuted tRNA genes in the photosynthetic eukaryotes implies that extant permuted tRNAs might have emerged multiple times. Taken together, our data demonstrate that permuted tRNA is an evolutionarily conserved and fundamental element in tiny eukaryotic genomes. PMID- 20022889 TI - POPTREE2: Software for constructing population trees from allele frequency data and computing other population statistics with Windows interface. AB - Currently, there is a demand for software to analyze polymorphism data such as microsatellite DNA and single nucleotide polymorphism with easily accessible interface in many fields of research. In this article, we would like to make an announcement of POPTREE2, a computer program package, that can perform evolutionary analyses of allele frequency data. The original version (POPTREE) was a command-line program that runs on the Command Prompt of Windows and Unix. In POPTREE2 genetic distances (measures of the extent of genetic differentiation between populations) for constructing phylogenetic trees, average heterozygosities (H) (a measure of genetic variation within populations) and G(ST) (a measure of genetic differentiation of subdivided populations) are computed through a simple and intuitive Windows interface. It will facilitate statistical analyses of polymorphism data for researchers in many different fields. POPTREE2 is available at http://www.med.kagawa-u.ac.jp/ approximately genomelb/takezaki/poptree2/index.html. PMID- 20022890 TI - Genome-wide association study in discordant sibships identifies multiple inherited susceptibility alleles linked to lung cancer. AB - We analyzed a series of young (median age = 52 years) non-smoker lung cancer patients and their unaffected siblings as controls, using a genome-wide 620 901 single-nucleotide polymorphism (SNP) array analysis and a case-control DNA pooling approach. We identified 82 putatively associated SNPs that were retested by individual genotyping followed by use of the sib transmission disequilibrium test, pointing to 36 SNPs associated with lung cancer risk in the discordant sibs series. Analysis of these 36 SNPs in a polygenic model characterized by additive and interchangeable effects of rare alleles revealed a highly statistically significant dosage-dependent association between risk allele carrier status and proportion of cancer cases. Replication of the same 36 SNPs in a population-based series confirmed the association with lung cancer for three SNPs, suggesting that phenocopies and genetic heterogeneity can play a major role in the complex genetics of lung cancer risk in the general population. PMID- 20022891 TI - The evolving discipline of molecular epidemiology of cancer. AB - Classical epidemiologic studies have made seminal contributions to identifying the etiology of most common cancers. Molecular epidemiology was conceived of as an extension of traditional epidemiology to incorporate biomarkers with questionnaire data to further our understanding of the mechanisms of carcinogenesis. Early molecular epidemiologic studies employed functional assays. These studies were hampered by the need for sequential and/or prediagnostic samples, viable lymphocytes and the uncertainty of how well these functional data (derived from surrogate lymphocytic tissue) reflected events in the target tissue. The completion of the Human Genome Project and Hapmap Project, together with the unparalleled advances in high-throughput genotyping revolutionized the practice of molecular epidemiology. Early studies had been constrained by existing technology to use the hypothesis-driven candidate gene approach, with disappointing results. Pathway analysis addressed some of the concerns, although the study of interacting and overlapping gene networks remained a challenge. Whole-genome scanning approaches were designed as agnostic studies using a dense set of markers to capture much of the common genome variation to study germ-line genetic variation as risk factors for common complex diseases. It should be possible to exploit the wealth of these data for pharmacogenetic studies to realize the promise of personalized therapy. Going forward, the temptation for epidemiologists to be lured by high-tech 'omics' will be immense. Systems Epidemiology, the observational prototype of systems biology, is an extension of classical epidemiology to include powerful new platforms such as the transcriptome, proteome and metabolome. However, there will always be the need for impeccably designed and well-powered epidemiologic studies with rigorous quality control of data, specimen acquisition and statistical analysis. PMID- 20022892 TI - Accuracy of cephalometric landmarks on monitor-displayed radiographs with and without image emboss enhancement. AB - The aim of this study was to evaluate the accuracy of some commonly used cephalometric landmarks of monitor-displayed images with and without image emboss enhancement. The following null hypothesis was tested: there is no improvement in landmark detection accuracy between monitor-displayed images, with and without image embossing enhancement. Forty lateral cephalometric radiographs, taken from the data files of subjects were used in this study. A purpose-made software allowed recording of the cephalometric points and then, with the help of algorithms based on cellular neural networks, to transfer the previously processed radiographs into an embossed image. Five observers recorded 22 landmarks on the displayed images from the two image modalities, i.e. monitor displayed radiograph (mode A) and monitor-displayed embossed radiograph (mode B). The positions of the landmarks were recorded and saved in the format of x and y co-ordinates and as Euclidean distance. The mean errors and standard deviation of landmarks location according to the two modalities were compared with the 'best estimate' for each landmark and the values were calculated for each of the 22 landmarks. One-way analysis of variance was then used to evaluate any statistically significant differences. Euclidean distance mean errors were higher for the embossed images (except for Po) than for the unfiltered radiographs. These differences were all statistically significant (P < 0.05) except for Or, Po, PM, Co, APOcc, and PPOcc. On the x and y co-ordinates, the accuracy of the cephalometric landmark detection improved on the embossed radiograph but only for a few points (Or on x axis and Po, PM, Co, and APOcc on y axis), as these were not statistically significant. The use of radiographic enhancement techniques, such as embossing, does not improve the level of accuracy for cephalometric points detection. Unless more precise algorithms are designed, this feature should not be used for clinical or research purposes. PMID- 20022893 TI - A prospective study of magnesium and iron intake and pancreatic cancer in men. AB - Many studies have investigated the relation between magnesium and iron intake and diabetes and, separately, between diabetes and pancreatic cancer. However, no known study has examined the direct association of magnesium and iron intake with pancreatic cancer risk. The authors obtained magnesium and iron intake data using food frequency questionnaires from the US male Health Professionals Follow-up Study, which began in 1986. During 851,476 person-years and 20 years of follow up, 300 pancreatic cancer cases were documented. Cox proportional hazards models were used to estimate relative risks, adjusting for age, smoking, and body mass index. No associations were observed between magnesium or iron intake and pancreatic cancer (highest vs. lowest quintile: relative risk (RR) = 0.94, 95% confidence interval (CI): 0.66, 1.32 and RR = 0.93, 95% CI: 0.65, 1.34, respectively). Similarly, iron or magnesium supplement use was not related to pancreatic cancer. A statistically significant inverse relation was noted between magnesium and pancreatic cancer for subjects with a body mass index of > or =25 kg/m(2) (RR = 0.67, 95% CI: 0.46, 0.99; P-trend = 0.04). Although, overall, no relation between magnesium or iron intake and pancreatic cancer was observed in this cohort of men, an inverse association with magnesium was suggested among overweight individuals, which should be examined in other studies. PMID- 20022894 TI - The potential for gene flow in a dependent lineage system of a harvester ant: fair meiosis in the F1 generation. AB - We investigated the potential for gene flow in a dependent lineage (DL) system of the harvester ant Pogonomyrmex. Each DL system is composed of 2 reproductively isolated lineages that are locked in an obligate mutualism. The genetic components that produce the worker phenotype are acquired by hybridizing with the partner lineage. In the mating flight, queens of both lineages mate with multiple males from each lineage. During colony growth and reproduction, eggs fertilized by partner-lineage sperm produce F(1) hybrid workers with interlineage genomes, whereas eggs fertilized by same-lineage sperm result in the development of new queens with intralineage genomes. New males are typically produced from unfertilized eggs laid by the pure-lineage queen but in her absence may be produced by interlineage F(1) workers. We investigated the potential for interlineage gene flow in this system using 2 classes of lineage-specific nuclear markers to identify hybrid genome combinations. We confirmed the production of viable interlineage F(1) reproductive females in field colonies, the occurrence of which is associated with the relative frequencies of each lineage in the population: interlineage F(1) queens occurred only in the rare lineage of the population with dramatically skewed lineage frequencies. In laboratory colonies, we detected fair meiosis in interlineage F(1) workers leading to the production of viable and haploid interlineage F(2) males. We conclude that the genomes of each lineage recombine freely, suggesting that extrinsic postzygotic selection maintains the integrity of each lineage genome. We compare our findings with those of the H1/H2 DL system. PMID- 20022895 TI - Biocharts: a visual formalism for complex biological systems. AB - We address one of the central issues in devising languages, methods and tools for the modelling and analysis of complex biological systems, that of linking high level (e.g. intercellular) information with lower-level (e.g. intracellular) information. Adequate ways of dealing with this issue are crucial for understanding biological networks and pathways, which typically contain huge amounts of data that continue to grow as our knowledge and understanding of a system increases. Trying to comprehend such data using the standard methods currently in use is often virtually impossible. We propose a two-tier compound visual language, which we call Biocharts, that is geared towards building fully executable models of biological systems. One of the main goals of our approach is to enable biologists to actively participate in the computational modelling effort, in a natural way. The high-level part of our language is a version of statecharts, which have been shown to be extremely successful in software and systems engineering. The statecharts can be combined with any appropriately well defined language (preferably a diagrammatic one) for specifying the low-level dynamics of the pathways and networks. We illustrate the language and our general modelling approach using the well-studied process of bacterial chemotaxis. PMID- 20022896 TI - Flow instability and wall shear stress variation in intracranial aneurysms. AB - We investigate the flow dynamics and oscillatory behaviour of wall shear stress (WSS) vectors in intracranial aneurysms using high resolution numerical simulations. We analyse three representative patient-specific internal carotid arteries laden with aneurysms of different characteristics: (i) a wide-necked saccular aneurysm, (ii) a narrower-necked saccular aneurysm, and (iii) a case with two adjacent saccular aneurysms. Our simulations show that the pulsatile flow in aneurysms can be subject to a hydrodynamic instability during the decelerating systolic phase resulting in a high-frequency oscillation in the range of 20-50 Hz, even when the blood flow rate in the parent vessel is as low as 150 and 250 ml min(-1) for cases (iii) and (i), respectively. The flow returns to its original laminar pulsatile state near the end of diastole. When the aneurysmal flow becomes unstable, both the magnitude and the directions of WSS vectors fluctuate at the aforementioned high frequencies. In particular, the WSS vectors around the flow impingement region exhibit significant spatio-temporal changes in direction as well as in magnitude. PMID- 20022897 TI - How necessary is a fast testkit for mitigation of pandemic flu? AB - It is widely feared that a novel, highly pathogenic, human transmissible influenza virus may evolve that could cause the next global pandemic. Mitigating the spread of such an influenza pandemic would require not only the timely administration of antiviral drugs to those infected, but also the implementation of suitable intervention policies for stunting the spread of the virus. Towards this end, mathematical modelling and simulation studies are crucial as they allow us to evaluate the predicted effectiveness of the various intervention policies before enforcing them. Diagnosis plays a vital role in the overall pandemic management framework by detecting and distinguishing the pathogenic strain from the less threatening seasonal strains and other influenza-like illnesses. This allows treatment and intervention to be deployed effectively, given limited antiviral supplies and other resources. However, the time required to design a fast and accurate testkit for novel strains may limit the role of diagnosis. Herein, we aim to investigate the cost and effectiveness of different diagnostic methods using a stochastic agent-based city-scale model, and then address the issue of whether conventional testing approaches, when used with appropriate intervention policies, can be as effective as fast testkits in containing a pandemic outbreak. We found that for mitigation purposes, fast and accurate testkits are not necessary as long as sufficient medication is given, and are generally recommended only when used with extensive contact tracing and prophylaxis. Additionally, in the event of insufficient medication and fast testkits, the use of slower, conventional testkits together with proper isolation policies while waiting for the diagnostic results can be an equally effective substitute. PMID- 20022898 TI - Mannose-binding lectin gene polymorphisms in Brazilian patients with systemic lupus erythematosus. AB - The mannose-binding lectin gene (MBL-2) has emerged as a candidate for systemic lupus erythematosus susceptibility, but studies in Brazilian population have not been conducted. To examine potential associations of mannose-binding lectin alleles G57E, G54D, IVSnt5, R52C and R52H with susceptibility to and clinical expression of systemic lupus erythematosus in southern Brazilian patients, we conducted a case-control study with 327 consecutive patients with diagnosis of systemic lupus erythematosus and 345 healthy controls. Genotyping was performed by restriction fragment length polymorphism-polymerase chain reaction assay. A statistically significant difference in the frequencies of allele R52C was observed in European-derived systemic lupus erythematosus patients when compared with controls (9.6% vs. 3.3%, p < 0.001, odds ratio: 3.15, 95% confidence interval: 1.76-5.62, p < 0.05). The frequencies of alleles G54D and G57E were not different between European-derived systemic lupus erythematosus patients and controls. There were no differences between clinical and laboratory features in systemic lupus erythematosus patients according to the presence or absence of mannose-binding lectin allelic variants. These results support an increased risk of systemic lupus erythematosus in European-derived individuals carrying allele R52C. Patients carrying this allele have an approximately three-fold higher odds ratio of developing systemic lupus erythematosus when compared with controls. Our data do not support associations between the mannose-binding lectin allelic variants studied and clinical expression of systemic lupus erythematosus. PMID- 20022899 TI - Residual household waste: from pay-per-bag to pay-per-kilogram. An evaluation study for Flanders. AB - To be in line with the polluter-pays principle, an increasing number of Flemish municipalities have introduced a weight-based pricing system for residual household waste. Municipalities using this new pricing scheme are keen to ascribe decreasing waste production to the introduction of the new system. In order to evaluate the real impact of the weight-based pricing, an extension to the standard difference-in-differences evaluation techniques is proposed. The main idea is to use historical observations on waste production to determine which version of the difference-in-differences technique is most suitable to assess the impact of a policy action on waste quantities. The method is easy to apply, which makes it a very useful tool for both scientists and policy makers. Surprisingly, empirical results for Flanders indicate that, on average, municipalities using the weight-based system do not perform better than other municipalities. PMID- 20022900 TI - Exploitation of biological wastes for the production of value-added hydrolases by Streptomyces sp. MSWC1 isolated from municipal solid waste compost. AB - Actinomycetes with the ability to degrade natural polysaccharides were isolated during a screening programme from soil, farmyard manure and municipal solid waste compost. One of the most potent isolates was identified as Streptomyces sp. MSWC1 using morphological and biochemical properties along with 16S rDNA partial sequence analysis. The highest enzyme production by Streptomyces was observed for the xylanase and chitinase activity on different carbon sources with an optimum of 12,100 IU ml(-1) and 110 IU ml(-1) at 3 days' culture on 1% of xylan and chitin, respectively. To meet the demand of industry, low-cost medium is required for the production of hydrolases by Streptomyces sp. Strain MSWC1 grown on manure, compost, and a natural carbon source was used to evaluate the re utilisation of biological wastes for the production of value-added products. Despite the presence of a high amount of toxic heavy metals in the compost, Streptomyces produced interesting enzymes that have been biochemically characterized. PMID- 20022901 TI - Optimisation research of petroleum hydrocarbon biodegradation in weathered drilling wastes from waste pits. AB - The aim of this article is to discuss the problem of drilling waste remediation. Analyses and research showed that material stored in waste pits could be classified as soil with a high level of petroleum impurities (total petroleum hydrocarbons [TPH] = 102,417-132,472 mg kg(-1) dry mass). While preparing the complex technology of soil decontamination (which included primary reclamation, basic bioremediation and inoculation with biopreparations based on indigenous bacteria and fungi), laboratory tests indicated the use of an ex-situ method was fundamental. Remediation was controlled with a chromatographic method of qualitative and quantitative determination of petroleum hydrocarbons. Based on analytical data, there was the possibility to determine the effectiveness of consecutive purifying phases. Laboratory tests, following 135 days of basic bioremediation stimulated by optimum conditions to activate the growth of indigenous micro-organisms, resulted in a decrease in the TPH content, which was in the range of 52.3-72.5%. The next phase of soil decontamination lasted 135 days and involved the use of inoculation with biopreparations based on indigenous micro-organisms and fungi. This process enabled a TPH decrease of 93.8- 94.3%. Laboratory biodegradation research was done with the use of the biomarker C30 17alpha(H)21beta(H)-hopane to normalize analyte (TPH, Sigman-C8-n-C22 and Sigman C23-n-C36) concentrations. The calculated first-order biodegradation constants enable estimation of the purification stage dynamics and the effectiveness of the applied biopreparations. Furthermore, they represent the biodegradation degree of individual n-alkanes in subsequent stages of the soil purification process. PMID- 20022902 TI - The pathway of in-situ ammonium removal from aerated municipal solid waste bioreactor: nitrification/denitrification or air stripping? AB - In-situ ammonium removal from municipal solid waste (MSW) landfill is an attractive method due to its economic advantages. In this study, two simulated MSW bioreactors with different degrees of initial bio-stabilization were utilized to investigate the effects of intermittent aeration mode and the addition of activated sludge on the removal of ammonium. The results showed that up to 90% of ammonium could be removed and the amount of NO(x)-N produced was less than 1% of NH4 (+)-N removed in both reactors. The pH values increased rapidly and finally arrived at a high level of 8.5-8.8. The efficiency of ammonium removal was improved by increasing the continuous aeration time, but it was not affected by the addition of activated sludge. A portion of liquid escaped from the reactors in the form of vapour, and as high as 195-258 mg L(-1) of NH(4) ( +)-N was detected in the vapour collector. According to calculation, nitrification was inhibited by the high level of free ammonia in the bioreactors. As a result, air stripping was enhanced and became the primary pathway of ammonium removal. Therefore, controlling free ammonia concentration was essential in ammonium removal from the aerated MSW bioreactor. PMID- 20022903 TI - Distribution of polycyclic aromatic hydrocarbons and polychlorinated dibenzo-p dioxins/dibenzofurans in ash from different units in a municipal solid waste incinerator. AB - Polycyclic aromatic hydrocarbons (PAHs), polychlorinated-p-dioxins (PCDDs) and polychlorinated dibenzofurans (PCDFs) are some of the major organic contaminants emitted in municipal solid waste incinerators (MSWIs). The distribution of PAHs and PCDD/Fs in different MSWI unit ashes, including bottom residue (BR), fly ash pit (FAP), fabric filter (FF), semi-dryer absorber (SDA), economizer (EC) and super heater (SH), has seldom been discussed. Average PAHs content in ash samples from BR, FAP, FF, SDA, EC and SH were found to be 0.020, 0.237, 0.234, 0.125, 0.037 and 0.034 ug TEQ g(-1), respectively. In addition, the range of total emission factor from different unit ashes varied between 0.30-13.85 mg TEQ tonne( 1) of waste, low molecular weight compounds (2~3-ring) being the dominant species. Furthermore, PAHs in ashes were mainly found in BR (41%) and FAP (31%). The FAP ash included 18% of PAHs coming from FF ash. On the other hand, PCDD/Fs content in BR, FAP, FF, SDA, EC and SH was found to be 0.0329, 1.1589, 1.2807, 0.1476, 0.6868 and 0.0921 ng I-TEQ g(-1), respectively. PCDD/ Fs emission factors in this study ranged between 1.029-67.740 ug I-TEQ tonne(-1) of waste, most PCDD/Fs being present in FAP (55%). The FAP ash included 33% of PCDD/Fs coming from FF ash. Therefore, PCDD/Fs content in FAP and FF exceeded the legal limit of 1 ng I-TEQ g( -1); therefore, a solidification process is necessary before landfilling in Taiwan. PMID- 20022904 TI - Editorial. Cairo: a colossal case of waste mismanagement to learn from. PMID- 20022906 TI - Analysis of cross-reactive and specific anti-carbohydrate antibodies against lipopolysaccharide from Chlamydophila psittaci. AB - Chlamydiae contain a rough-type lipopolysaccharide (LPS) of 3-deoxy-alpha-d-manno oct-2-ulopyranosonic acid residues (Kdo). Two Kdo trisaccharides, 2.8/2.4- and 2.4/2.4-linked, and a branched 2.4[2.8]2.4-linked Kdo tetrasaccharide occur in Chlamydiaceae. While the 2.8/2.4-linked trisaccharide contains a family-specific epitope, the branched Kdo oligosaccharide occurs only in Chlamydophila psittaci and antibodies against it will be useful in human and veterinarian diagnostics. To overcome the generation of cross-reactive antibodies that bind with high affinity to a dominant epitope formed by 2.4/2.4-linked Kdo, we immunized mice with a synthetic 2.4[2.8]-linked branched Kdo trisaccharide and used phage display of scFv to isolate recombinant antibody fragments (NH2240-31 and SAG506 01) that recognize the branched Kdo oligosaccharide with a K(D) of less than 10 nM. Importantly, although these antibodies used germline genes coding for an inherited Kdo recognition site, they were able clearly to distinguish between 2.4[2.8]2.4- and 2.4/2.4-linked Kdo. Sequence determination, binding data, and X ray structural analysis revealed the basis for the improved discrimination between similar Kdo ligands and indicated that the alteration of a stacking interaction from a phenylalanine residue in the center of the combining site to a tyrosine residue facing away from the center favors recognition of branched 2.4[2.8]2.4-linked Kdo residues. Immunofluorescence tests of infected cell monolayers using this antibody show specific staining of C. psittaci elementary bodies that allow it to be distinguished from other pathogenic chlamydiae. PMID- 20022907 TI - Information in general medical practices: the information processing model. AB - BACKGROUND: The need for effective communication and handling of secondary care information in general practices is paramount. OBJECTIVE: To explore practice processes on receiving secondary care correspondence in a way that integrates the information needs and perceptions of practice staff both clinical and administrative. METHODS: Qualitative study using semi-structured interviews with a wide range of practice staff (n = 36) in nine practices in the Northwest of England. Analysis was based on the framework approach using N-Vivo software and involved transcription, familiarization, coding, charting, mapping and interpretation. RESULTS: The 'information processing model' was developed to describe the six stages involved in practice processing of secondary care information. These included the amendment or updating of practice records whilst simultaneously or separately actioning secondary care recommendations, using either a 'one-step' or 'two-step' approach, respectively. Many factors were found to influence each stage and impact on the continuum of patient care. CONCLUSION: The primary purpose of processing secondary care information is to support patient care; this study raises the profile of information flow and usage within practices as an issue requiring further consideration. PMID- 20022908 TI - Communication challenges of HPV vaccination. AB - BACKGROUND: The UK has introduced a national human papillomavirus (HPV) immunisation programme for girls aged 12-13 from autumn 2008. Previous research has explored public understanding of the causes of cervical cancer and found little awareness of the role of HPV. OBJECTIVE: To explore GPs' and practice nurses' views of HPV vaccination, prior to implementation of the national immunisation programme, with a focus on their role and anticipated difficulties. METHODS: Qualitative semi-structured interviews were conducted with 17 GPs and practice nurses from practices in Hampshire and Wiltshire, in March 2008. RESULTS: The prevailing theme of new communication challenges was identified and these were connected to the activities of (i) explaining, (ii) consenting and (iii) managing conflicts between parents and their children with respect to vaccination decisions. The importance of decisions being fully informed was emphasized and concern was expressed about the adequacy of the information provided to girls and their parents in schools. Whether consent would be granted by the parent or by the child and how potential disagreements should be managed remained ambiguous. Participants considered it appropriate to offer the vaccine without parental consent if other criteria, such as an assessment of competency, were met. CONCLUSIONS: Success of the national immunization programme will depend on overcoming the challenges of providing explanations to ensure that individuals understand the potential benefits of HPV vaccination. Primary care can play an important role, but this study suggests there are fundamental issues that need further clarification. PMID- 20022909 TI - C-reactive protein point-of-care testing for lower respiratory tract infections: a qualitative evaluation of experiences by GPs. AB - OBJECTIVES: To explore GPs' attitudes to and experiences of introducing C reactive protein (CRP) point-of-care testing (POCT) for lower respiratory tract infections (LRTI) in primary care. METHODS: Semi-structured interview study with 20 GPs who participated in the IMPAC(3)T randomized trial evaluating the effect of GP use of CRP POCT on management of LRTI. Main outcomes were GPs' experiences and views about CRP POCT in general practice, including its role in guiding antibiotic prescribing decisions and applicability and implementation in daily practice. RESULTS: GPs expressed mainly positive attitudes. Test results were rapidly available to support diagnostic and therapeutic processes for LRTI and other common infections, enhancing patient and GP confidence in prescribing decisions and empowering GPs to prescribe antibiotics less often. GPs were concerned about responding to ambiguous test results. They regarded financial reimbursement for using the test as essential for successful uptake. CONCLUSIONS: GPs were generally positive about CRP POCT, and they felt that it empowered them to safely prescribe fewer antibiotics for LRTI without alienating their patients. Successful wider implementation should address reimbursement and updating management guidelines to include the place of CRP POCT. PMID- 20022910 TI - The indolent course and high incidence of t(14;18) in primary duodenal follicular lymphoma. AB - BACKGROUND: Information on the clinical behavior of the recently proposed primary duodenal follicular lymphoma (DFL) is limited. PATIENTS AND METHODS: Demographic data, signs, symptoms, disease stage, and treatment of the patients diagnosed in National Cancer Center Hospital from 1999 to 2007 were collected and analyzed. RESULTS: Twenty-seven patients were studied. Nineteen patients were asymptomatic at the time of diagnosis. Twenty patients had stage I disease. The histological grade was 1 or 2 in 26 patients. IgH/BCL2 fusion was shown in 20 of the examined 24 cases (83%). Fourteen patients received therapy upon diagnosis (local radiotherapy in 2 patients and chemotherapy in 12 including rituximab therapy), their response rate was 85%, and the estimated progression-free survival (PFS) rate at 3 years was 70%. One patient developed histological transformation. The other 13 patients were followed up; their estimated PFS rate at 3 years was 74%. Five among six cases responded to treatment even after progressive disease. All 27 patients have survived with a median follow-up time of 47.9 months. CONCLUSIONS: The majority of primary DFL patients have a localized tumor of low grade histology and are positive for t(14;18). Watchful waiting might be an alternative approach for its indolent course; however, further studies are warranted. PMID- 20022911 TI - Two-protein signature of novel serological markers apolipoprotein-A2 and serum amyloid alpha predicts prognosis in patients with metastatic renal cell cancer and improves the currently used prognostic survival models. AB - BACKGROUND: In metastatic renal cell cancer (mRCC), the Memorial Sloan-Kettering Cancer Center (MSKCC) risk model is widely used for clinical trial design and patient management. To improve prognostication, we applied proteomics to identify novel serological proteins associated with overall survival (OS). PATIENTS AND METHODS: Sera from 114 mRCC patients were screened by surface-enhanced laser desorption ionization time-of-flight mass spectrometry (SELDI-TOF MS). Identified proteins were related to OS. Three proteins were subsequently validated with enzyme-linked immunosorbent assays and immunoturbidimetry. Prognostic models were statistically bootstrapped to correct for overestimation. RESULTS: SELDI-TOF MS detected 10 proteins associated with OS. Of these, apolipoprotein A2 (ApoA2), serum amyloid alpha (SAA) and transthyretin were validated for their association with OS (P = 5.5 x 10(-9), P = 1.1 x 10(-7) and P = 0.0004, respectively). Combining ApoA2 and SAA yielded a prognostic two-protein signature [Akaike's Information Criteria (AIC) = 732, P = 5.2 x 10(-7)]. Including previously identified prognostic factors, multivariable Cox regression analysis revealed ApoA2, SAA, lactate dehydrogenase, performance status and number of metastasis sites as independent factors for survival. Using these five factors, categorization of patients into three risk groups generated a novel protein-based model predicting patient prognosis (AIC = 713, P = 4.3 x 10(-11)) more robustly than the MSKCC model (AIC = 729, P = 1.3 x 10(-7)). Applying this protein-based model instead of the MSKCC model would have changed the risk group in 38% of the patients. CONCLUSIONS: Proteomics and subsequent validation yielded two novel prognostic markers and survival models which improved prediction of OS in mRCC patients over commonly used risk models. Implementation of these models has the potential to improve current risk stratification, although prospective validation will still be necessary. PMID- 20022912 TI - Microbicide offers no protection against HIV infection. PMID- 20022913 TI - The molecular receptive ranges of human TAS2R bitter taste receptors. AB - Humans perceive thousands of compounds as bitter. In sharp contrast, only approximately 25 taste 2 receptors (TAS2R) bitter taste receptors have been identified, raising the question as to how the vast array of bitter compounds can be detected by such a limited number of sensors. To address this issue, we have challenged 25 human taste 2 receptors (hTAS2Rs) with 104 natural or synthetic bitter chemicals in a heterologous expression system. Thirteen cognate bitter compounds for 5 orphan receptors and 64 new compounds for previously identified receptors were discovered. Whereas some receptors recognized only few agonists, others displayed moderate or extreme tuning broadness. Thus, 3 hTAS2Rs together were able to detect approximately 50% of the substances used. Conversely, though 63 bitter substances activated only 1-3 receptors, 19 compounds stimulated up to 15 hTAS2Rs. Our data suggest that the detection of the numerous bitter chemicals is related to the molecular receptive ranges of hTAS2Rs. PMID- 20022914 TI - Impact of forced separation policy on incarcerated postpartum mothers. AB - The number of incarcerated women in the United States is steadily increasing with 9% giving birth while serving time. Mothers and babies are routinely separated immediately and during most of the postpartum period. This qualitative study examines the impact of this policy by exploring the nature and meaning of the mother-infant bonding experience when the mothers know separation is coming. Twelve incarcerated postpartum mothers were interviewed during the early postpartum period about their antepartum and postpartum relationships with their babies. Qualitative data analysis produced four relational themes: (a) "a love connection," (b) "everything was great until I birthed," (c) "feeling empty and missing a part of me," and (d) "I don't try to think too far in advance." Results of this qualitative study provide information to professionals exploring gender sensitive prison policies for the growing female population. PMID- 20022915 TI - Do wages matter?: a backward bend in the 2004 California RN labor supply. AB - Using data from the 2004 California Board of Registered Nursing Survey, a two stage least-square equation was estimated to examine the effect of wages on hours worked by female registered nurses. Wages were found to have a nonlinear effect on hours worked, with a backward bending supply curve. Wages had a positive effect on the average hours worked per week up to $24.99 per hour and a negative effect between $30.00 and $100.00 per hour when compared with the wage category of $25.00 to $29.99. Results suggest that wages are important to secure the labor supply but do not increase aggregate supply beyond a wage threshold. PMID- 20022916 TI - Riboswitch structure: an internal residue mimicking the purine ligand. AB - The adenine and guanine riboswitches regulate gene expression in response to their purine ligand. X-ray structures of the aptamer moiety of these riboswitches are characterized by a compact fold in which the ligand forms a Watson-Crick base pair with residue 65. Phylogenetic analyses revealed a strict restriction at position 39 of the aptamer that prevents the G39-C65 and A39-U65 combinations, and mutational studies indicate that aptamers with these sequence combinations are impaired for ligand binding. In order to investigate the rationale for sequence conservation at residue 39, structural characterization of the U65C mutant from Bacillus subtilis pbuE adenine riboswitch aptamer was undertaken. NMR spectroscopy and X-ray crystallography studies demonstrate that the U65C mutant adopts a compact ligand-free structure, in which G39 occupies the ligand-binding site of purine riboswitch aptamers. These studies present a remarkable example of a mutant RNA aptamer that adopts a native-like fold by means of ligand mimicking and explain why this mutant is impaired for ligand binding. Furthermore, this work provides a specific insight into how the natural sequence has evolved through selection of nucleotide identities that contribute to formation of the ligand-bound state, but ensures that the ligand-free state remains in an active conformation. PMID- 20022917 TI - Generation and bioenergetic analysis of cybrids containing mitochondrial DNA from mouse skeletal muscle during aging. AB - Mitochondrial respiratory chain defects have been associated with various diseases and normal aging, particularly in tissues with high energy demands including skeletal muscle. Muscle-specific mitochondrial DNA (mtDNA) mutations have also been reported to accumulate with aging. Our understanding of the molecular processes mediating altered mitochondrial gene expression to dysfunction associated with mtDNA mutations in muscle would be greatly enhanced by our ability to transfer muscle mtDNA to established cell lines. Here, we report the successful generation of mouse cybrids carrying skeletal muscle mtDNA. Using this novel approach, we performed bioenergetic analysis of cells bearing mtDNA derived from young and old mouse skeletal muscles. A significant decrease in oxidative phosphorylation coupling and regulation capacity has been observed with cybrids carrying mtDNA from skeletal muscle of old mice. Our results also revealed decrease growth capacity and cell viability associated with the mtDNA derived from muscle of old mice. These findings indicate that a decline in mitochondrial function associated with compromised mtDNA quality during aging leads to a decrease in both the capacity and regulation of oxidative phosphorylation. PMID- 20022918 TI - Engineering of stable bispecific antibodies targeting IL-17A and IL-23. AB - Bispecific antibodies (bsAbs) present an attractive opportunity to combine the additive and potentially synergistic effects exhibited by combinations of monoclonal antibodies (mAbs). Current challenges for engineering bsAbs include retention of the binding affinity of the parent mAb or antibody fragment, the ability to bind both targets simultaneously, and matching valency with biology. Other factors to consider include structural stability and expression of the recombinant molecule, both of which may have significant impact on its development as a therapeutic. Here, we incorporate selection of stable, potent single-chain variable fragments (scFvs) early in the engineering process to assemble bsAbs for therapeutic applications targeting the cytokines IL-17A/A and IL-23. Stable scFvs directed against human cytokines IL-23p19 and IL-17A/A were isolated from a human Fab phage display library via batch conversion of panning output from Fabs to scFvs. This strategy integrated a step for shuffling V regions during the conversion and permitted the rescue of scFv molecules in both the V(H)V(L) and the V(L)V(H) orientations. Stable scFvs were identified and assembled into several bispecific formats as fusions to the Fc domain of human IgG1. The engineered bsAbs are potent neutralizers of the biological activity of both cytokines (IC(50) < 1 nM), demonstrate the ability to bind both target ligands simultaneously and display stability and productivity advantageous for successful manufacture of a therapeutic molecule. Pharmacokinetic analysis of the bsAbs in mice revealed serum half-lives similar to human mAbs. Assembly of bispecific molecules using stable antibody fragments offers an alternative to reformatting mAbs and minimizes subsequent structure-related and manufacturing concerns. PMID- 20022919 TI - Long-term physical health consequences of childhood sexual abuse: a meta-analytic review. AB - OBJECTIVE: The purpose of the present article was to systematically review the literature investigating the long-term physical health consequences of childhood sexual abuse (CSA). METHODS: Literature searches yielded 31 studies comparing individuals with and without a history of CSA on six health outcomes: general health, gastrointestinal (GI) health, gynecologic or reproductive health, pain, cardiopulmonary symptoms, and obesity. Exploratory subgroup analyses were conducted to identify potential methodological moderators. RESULTS: Results suggested that a history of CSA was associated with small to moderate group differences on almost all health outcomes assessed, such that individuals with a history of CSA reported more complaints for each health outcome. Suggestive trends in moderating variables of study design and methodology are presented. CONCLUSIONS: Results highlight the long-term physical health consequences of CSA and identify potential moderators to aid in the design of future research. PMID- 20022920 TI - Thermoperiodic growth control by gibberellin does not involve changes in photosynthetic or respiratory capacities in pea. AB - Active gibberellin (GA(1)) is an important mediator of thermoperiodic growth in pea. Plants grown under lower day than night temperature (negative DIF) elongate less and have reduced levels of GA(1) compared with plants grown at higher day than night temperature (positive DIF). By comparing the wild type (WT) and the elongated DELLA mutant la cry(s), this study has examined the effect of impaired GA signalling on thermoperiodic growth, photosynthesis, and respiration in pea. In the WT a negative DIF treatment reduced stem mass ratio and increased both root mass ratio and leaf mass ratio (dry weight of specific tissue related to total plant dry weight). Leaf, root and stem mass ratios of la cry(s) were not affected by DIF. Under negative DIF, specific leaf area (projected leaf area per unit leaf dry mass), biomass, and chlorophyll content of WT and la cry(s) plants were reduced. Young, expanding leaves of plants grown under negative DIF had reduced leaf area-based photosynthetic capacity. However, the highest photosynthetic electron transport rate was found in fully expanded leaves of WT plants grown under negative DIF. Negative DIF increased night respiration and was similar for both genotypes. It is concluded that GA signalling is not a major determinant of leaf area-based photosynthesis or respiration and that reduced dry weight of plants grown under negative DIF is caused by a GA-mediated reduction of photosynthetic stem and leaf tissue, reduced photosynthesis of young, expanding leaves, and reduced growth caused by low temperature in the photoperiod. PMID- 20022921 TI - TaSnRK2.4, an SNF1-type serine/threonine protein kinase of wheat (Triticum aestivum L.), confers enhanced multistress tolerance in Arabidopsis. AB - Osmotic stresses such as drought, salinity, and cold are major environmental factors that limit agricultural productivity worldwide. Protein phosphorylation/dephosphorylation are major signalling events induced by osmotic stress in higher plants. Sucrose non-fermenting 1-related protein kinase2 family members play essential roles in response to hyperosmotic stresses in Arabidopsis, rice, and maize. In this study, the function of TaSnRK2.4 in drought, salt, and freezing stresses in Arabidopsis was characterized. A translational fusion protein of TaSnRK2.4 with green fluorescent protein showed subcellular localization in the cell membrane, cytoplasm, and nucleus. To examine the role of TaSnRK2.4 under various environmental stresses, transgenic Arabidopsis plants overexpressing wheat TaSnRK2.4 under control of the cauliflower mosaic virus 35S promoter were generated. Overexpression of TaSnRK2.4 resulted in delayed seedling establishment, longer primary roots, and higher yield under normal growing conditions. Transgenic Arabidopsis overexpressing TaSnRK2.4 had enhanced tolerance to drought, salt, and freezing stresses, which were simultaneously supported by physiological results, including decreased rate of water loss, enhanced higher relative water content, strengthened cell membrane stability, improved photosynthesis potential, and significantly increased osmotic potential. The results show that TaSnRK2.4 is involved in the regulation of enhanced osmotic potential, growth, and development under both normal and stress conditions, and imply that TaSnRK2.4 is a multifunctional regulatory factor in Arabidopsis. Since the overexpression of TaSnRK2.4 can significantly strengthen tolerance to drought, salt, and freezing stresses and does not retard the growth of transgenic Arabidopsis plants under well-watered conditions, TaSnRK2.4 could be utilized in transgenic breeding to improve abiotic stresses in crops. PMID- 20022922 TI - The tobacco MAP215/Dis1-family protein TMBP200 is required for the functional organization of microtubule arrays during male germline establishment. AB - The haploid microspore division during pollen development in flowering plants is an intrinsically asymmetric division which establishes the male germline for sexual reproduction. Arabidopsis gem1 mutants lack the male germline as a result of disturbed microspore polarity, division asymmetry, and cytokinesis and represent loss-of-function mutants in MOR1/GEM1, a plant orthologue of the conserved MAP215/Dis1 microtubule associated protein (MAP) family. This provides genetic evidence for the role of MAP215/Dis1 in the organization of gametophytic microtubule arrays, but it has remained unknown how microtubule arrays are affected in gem1 mutant microspores. Here, novel male gametophytic microtubule reporter Nicotiana tabacum plants were constructed, expressing a green fluorescent protein-alpha-TUBULIN fusion protein (GFP-TUA6) under the control of a microspore-specific promoter. These plants allow effective visualization of all major male gametophytic microtubule arrays and provide useful tools to study the regulation of microtubule arrays by MAPs and other effectors. Depletion of TMBP200, a tobacco homologue of MOR1/GEM1 in gametophytic microtubule-reporter plants using microspore-targeted RNA interference, induced defects in microspore polarity, division asymmetry and cytokinesis that were associated with striking defects in phragmoplast position, orientation, and structure. Our observations further reveal a requirement for TMBP200 in gametophytic spindle organization and a novel role in spindle position and orientation in polarized microspores. These results provide direct evidence for the function of MAP215/Dis1 family protein TMBP200 in the organization of microtubule arrays critical for male germline formation in plants. PMID- 20022923 TI - Redox states of glutathione and ascorbate in root tips of poplar (Populus tremula X P. alba) depend on phloem transport from the shoot to the roots. AB - Glutathione (GSH) and ascorbate (ASC) are important antioxidants that are involved in stress defence and cell proliferation of meristematic root cells. In principle, synthesis of ASC and GSH in the roots as well as ASC and GSH transport from the shoot to the roots by phloem mass flow is possible. However, it is not yet known whether the ASC and/or the GSH level in roots depends on the supply from the shoot. This was analysed by feeding mature leaves with [(14)C]ASC or [(35)S]GSH and subsequent detection of the radiolabel in different root fractions. Quantitative dependency of root ASC and GSH on shoot-derived ASC and GSH was investigated with poplar (Populus tremula X P. alba) trees interrupted in phloem transport. [(35)S]GSH is transported from mature leaves to the root tips, but is withdrawn from the phloem along the entire transport path. When phloem transport was interrupted, the GSH content in root tips halved within 3 d. [(14)C]ASC is also transported from mature leaves to the root tips but, in contrast to GSH, ASC is not removed from the phloem along the transport path. Accordingly, ASC accumulates in root tips. Interruption of phloem transport disturbed the level and the ASC redox state within the entire root system. Diminished total ASC levels were attributed mainly to a decline of dehydroascorbate (DHA). As the redox state of ASC is of particular significance for root growth and development, it is concluded that phloem transport of ASC may constitute a shoot to root signal to coordinate growth and development at the whole plant level. PMID- 20022924 TI - A major quantitative trait locus for increasing cadmium-specific concentration in rice grain is located on the short arm of chromosome 7. AB - Large phenotypic variations in the cadmium (Cd) concentration of rice grains and shoots have been observed. However, the genetic control of Cd accumulation remains poorly understood. Quantitative trait loci (QTLs) determining the grain Cd concentration of rice grown in a Cd-polluted paddy field were identified. Using a mapping population consisting of 85 backcross inbred lines derived from a cross between the low-Cd-accumulating cultivar Sasanishiki (japonica) and high-Cd accumulating cultivar Habataki (indica), two QTLs for increasing grain Cd concentration were found on chromosomes 2 and 7. A major-effect QTL, qGCd7 (QTL for grain Cd on chromosome 7), was detected on the short arm of chromosome 7. It accounted for 35.5% of all phenotypic variance in backcross inbred lines. qGCd7 was not genetically related to any QTLs for concentrations of essential trace metals (Cu, Fe, Mn, and Zn) or those for agronomic traits such as heading date, suggesting that this QTL is specific to Cd. Furthermore, the existence of qGCd7 was confirmed using chromosome segment substitution lines (CSSLs) and an F(2) population from a cross between the target CSSL and Sasanishiki grown in a Cd polluted paddy soil. To our knowledge, qGCd7 is a novel QTL with major effects for increasing grain Cd concentrations. PMID- 20022925 TI - The life course and the stress process: some conceptual comparisons. AB - This paper compares the meanings and applications of concepts relevant to both the life course and the stress process frameworks. Some of these concepts bear the same labels but serve quite different scholarly agendas. Other concepts have different labels but have closely related applications. The purpose of this kind of comparative analysis is to help both fields clarify the conceptual tools needed to advance their scholarly goals. PMID- 20022926 TI - Sexual behaviours and the risk of head and neck cancers: a pooled analysis in the International Head and Neck Cancer Epidemiology (INHANCE) consortium. AB - BACKGROUND: Sexual contact may be the means by which head and neck cancer patients are exposed to human papillomavirus (HPV). METHODS: We undertook a pooled analysis of four population-based and four hospital-based case-control studies from the International Head and Neck Cancer Epidemiology (INHANCE) consortium, with participants from Argentina, Australia, Brazil, Canada, Cuba, India, Italy, Spain, Poland, Puerto Rico, Russia and the USA. The study included 5642 head and neck cancer cases and 6069 controls. We calculated odds ratios (ORs) of associations between cancer and specific sexual behaviours, including practice of oral sex, number of lifetime sexual partners and oral sex partners, age at sexual debut, a history of same-sex contact and a history of oral-anal contact. Findings were stratified by sex and disease subsite. RESULTS: Cancer of the oropharynx was associated with having a history of six or more lifetime sexual partners [OR = 1.25, 95% confidence interval (CI) 1.01, 1.54] and four or more lifetime oral sex partners (OR = 2.25, 95% CI 1.42, 3.58). Cancer of the tonsil was associated with four or more lifetime oral sex partners (OR = 3.36, 95 % CI 1.32, 8.53), and, among men, with ever having oral sex (OR = 1.59, 95% CI 1.09, 2.33) and with an earlier age at sexual debut (OR = 2.36, 95% CI 1.37, 5.05). Cancer of the base of the tongue was associated with ever having oral sex among women (OR = 4.32, 95% CI 1.06, 17.6), having two sexual partners in comparison with only one (OR = 2.02, 95% CI 1.19, 3.46) and, among men, with a history of same-sex sexual contact (OR = 8.89, 95% CI 2.14, 36.8). CONCLUSIONS: Sexual behaviours are associated with cancer risk at the head and neck cancer subsites that have previously been associated with HPV infection. PMID- 20022927 TI - Cortisol, estrogens and risk of ischaemic heart disease, cancer and all-cause mortality in postmenopausal women: a prospective cohort study. AB - BACKGROUND: Chronic stress may affect morbidity and mortality through neuroendocrine changes, and the ratio of cortisol to sex steroid hormones has been suggested as a biomarker of stress. We aim to address a relation between the ratio of cortisol to estrogens (C/E) and risk of ischaemic heart disease (IHD), hormone-dependent cancers and all-cause mortality among postmenopausal women. METHODS: Estradiol and cortisol were measured in a subset of 838 women randomly drawn from the second wave of the Copenhagen City Heart Study (n = 5297 examined in 1981-83) as well as among all women who developed hormone-dependent cancers after baseline. The participants were followed in nationwide registers until 2007, with < 0.1% loss to follow-up. RESULTS: The C/E ratio was not associated with self-reported stress, and there were no clear associations with IHD (highest vs lowest quartile: HR = 1.23; 95% confidence interval 0.83-1.81), hormone dependent cancers (0.69; 0.45-1.08) or all-cause mortality (1.10; 0.86-1.41). CONCLUSIONS: The C/E ratio was not associated with morbidity and mortality in women, and we cannot replicate the robust findings of a relation between the cortisol to testosterone ratio and IHD previously reported in men. Whether the C/E ratio is a reasonable biomarker of stress is debatable. PMID- 20022928 TI - A potential cause of the reported increase in rates of autism. PMID- 20022929 TI - Testosterone supplementation reverses sarcopenia in aging through regulation of myostatin, c-Jun NH2-terminal kinase, Notch, and Akt signaling pathways. AB - Aging in rodents and humans is characterized by loss of muscle mass (sarcopenia). Testosterone supplementation increases muscle mass in healthy older men. Here, using a mouse model, we investigated the molecular mechanisms by which testosterone prevents sarcopenia and promotes muscle growth in aging. Aged mice of 22 months of age received a single sc injection of GnRH antagonist every 2 wk to suppress endogenous testosterone production and were implanted subdermally under anesthesia with 0.5 or 1.0 cm testosterone-filled implants for 2 months (n = 15/group). Young and old mice (n = 15/group), of 2 and 22 months of age, respectively, received empty implants and were used as controls. Compared with young animals, a significant (P < 0.05) increase in muscle cell apoptosis coupled with a decrease in gastrocnemius muscles weight (by 16.7%) and muscle fiber cross sectional area, of both fast and slow fiber types, was noted in old mice. Importantly, such age-related changes were fully reversed by higher dose (1 cm) of testosterone treatment. Testosterone treatment effectively suppressed age specific increases in oxidative stress, processed myostatin levels, activation of c-Jun NH(2)-terminal kinase, and cyclin-dependent kinase inhibitor p21 in aged muscles. Furthermore, it restored age-related decreases in glucose-6-phosphate dehydrogenase levels, phospho-Akt, and Notch signaling. These alterations were associated with satellite cell proliferation and differentiation. Collectively these results suggest involvement of multiple signal transduction pathways in sarcopenia. Testosterone reverses sarcopenia through stimulation of cellular metabolism and survival pathway together with inhibition of death pathway. PMID- 20022930 TI - Endoplasmic reticulum stress-induced activation of activating transcription factor 6 decreases cAMP-stimulated hepatic gluconeogenesis via inhibition of CREB. AB - The expression of genes encoding key hepatic gluconeogenic enzymes, including phosphoenolpyruvate carboxykinase (PEPCK) and glucose-6-phosphatase (G6Pase), is regulated at the transcriptional level by a network of transcription factors and cofactors, including cAMP response element-binding protein (CREB). It has been suggested that increased endoplasmic reticulum (ER) stress in the liver impairs hepatic glucose metabolism. However, the direct effect of ER stress on hepatic gluconeogenesis is still not clear. Here, we investigated whether ER stress influences hepatic gluconeogenesis and whether this process is mediated by activating transcription factor 6 (ATF6) through the inhibition of cAMP-mediated activation of CREB. A cAMP stimulant, forskolin, and 8-bromoadenosine-cAMP increased PEPCK and G6Pase mRNA expression in H4IIE rat hepatoma cells, and ER stress induced by tunicamycin or thapsigargin decreased the expression of these genes in forskolin or 8-bromoadenosine-cAMP-treated cells. In a transient transfection study, ATF6 inhibited the PEPCK and G6Pase promoters. Also, adenovirus-mediated overexpression of ATF6 in H4IIE cells decreased forskolin stimulated PEPCK and G6Pase gene expression. Moreover, the inhibition of endogenous ATF6 expression by small interfering RNAs restored the ER stress induced suppression of PEPCK and G6Pase gene expression. Transient transfection of ATF6 inhibited transactivation by CREB on the PEPCK and G6Pase promoters, and a gel shift assay showed that Ad-ATF6 inhibits forskolin-stimulated CREB DNA binding activity. Finally, we found that expression of ATF6 decreased fasting induced PEPCK, G6Pase mRNA expression, and blood glucose levels in mice. Taken together, these data extend our understanding of ER stress and the regulation of liver gluconeogenesis by ATF6. PMID- 20022931 TI - Role of asparagine-linked glycosylation in cell surface expression and function of the human adrenocorticotropin receptor (melanocortin 2 receptor) in 293/FRT cells. AB - Asparagine-linked glycosylation (N-glycosylation) of G protein-coupled receptors may be necessary for functions ranging from agonist binding, folding, maturation, stability, and internalization. Human melanocortin 2 receptor (MC2R) possesses putative N-glycosylation sites in its N-terminal extracellular domain; however, to date, the role of MC2R N-glycosylation has yet to be investigated. The objective of the present study is to examine whether N-glycosylation is essential or not for cell surface expression and cAMP production in native and MC2R accessory protein (MRAP alpha, -beta, or -dCT)-expressing cells using 293/FRT transfected with Myc-MC2R. Western blot analyses performed with or without endoglycosidase H, peptide:N-glycosidase F or tunicamycin treatments and site directed mutagenesis revealed that MC2R was glycosylated in the N-terminal domain at its two putative N-glycosylation sites (Asn(12)-Asn(13)-Thr(14) and Asn(17) Asn(18)-Ser(19)). In the absence of human MRAP coexpression, N-glycosylation of at least one of the two sites was necessary for MC2R cell surface expression. However, when MRAP was present, cell surface expression of MC2R mutants was either rescued entirely with the N17-18Q (QQNN) and N12-13Q (NNQQ) mutants or partially with the unglycosylated N12-13, 17-18Q (QQQQ) mutant. Functional and expression analyses revealed a discrepancy between wild-type (WT) and QQQQ cell surface receptor levels and maximal cAMP production with a 4-fold increase in EC(50) values. Taken together, these results indicate that the absence of MC2R N glycosylation abrogates to a large extent MC2R cell surface expression in the absence of MRAPs, whereas when MC2R is N-glycosylated, it can be expressed at the plasma membrane without MRAP assistance. PMID- 20022932 TI - Cyclooxygenase 2 inhibition exacerbates palmitate-induced inflammation and insulin resistance in skeletal muscle cells. AB - Palmitate-induced inflammation is involved in the development of insulin resistance in skeletal muscle cells. Here we evaluated the effect of the saturated fatty acid palmitate and the monounsaturated fatty acid oleate on Toll like receptors (TLR)-2 and -4 and cyclooxygenase 2 (COX-2) expression and examined whether the inhibition of this enzyme modulates fatty acid-induced inflammation. Skeletal muscle cells exposed to palmitate showed enhanced TLR-2 and COX-2 mRNA levels, whereas oleate did not modify their expression. Palmitate induced expression of these genes was dependent on nuclear factor (NF)-kappaB activation, because expression was reduced in the presence of the NF-kappaB inhibitor parthenolide. Coincubation of palmitate-exposed cells with oleate also prevented the increase in the expression of TLR-2 and COX-2, through a mechanism that may involve activation of peroxisome proliferator-activated receptor-alpha (PPAR alpha) by this monounsaturated fatty acid. COX-2 inhibition by NS-398 enhanced IL-6 and TNF-alpha expression and IL-6 protein secretion induced by palmitate. NF-kappaB binding activity and TNF-alpha mRNA levels were enhanced in palmitate-exposed cells in the absence or in the presence of NS-398, whereas coincubation of palmitate-exposed cells with NS-398 and prostaglandin E(2) (PGE(2)) prevented these changes. In contrast, 12-lypoxygenase and cytochrome P450 hydroxylase pathways were not involved in these changes. Similarly, COX-2 inhibition impaired insulin-stimulated Akt phosphorylation and 2-deoxy-D [(14)C]glucose uptake in palmitate-exposed skeletal muscle cells, and this effect was abolished in the presence of PGE(2). These findings indicate that COX-2 activity, through the production of PGE(2), attenuates the fatty acid-induced inflammatory process and insulin resistance. PMID- 20022933 TI - Plasma transcortin influences endocrine and behavioral stress responses in mice. AB - Glucocorticoids are released after hypothalamus-pituitary-adrenal axis stimulation by stress and act both in the periphery and in the brain to bring about adaptive responses that are essential for life. Dysregulation of the stress response can precipitate psychiatric diseases, in particular depression. Recent genetic studies have suggested that the glucocorticoid carrier transcortin, also called corticosteroid-binding globulin (CBG), may have an important role in stress response. We have investigated the effect of partial or total transcortin deficiency using transcortin knockout mice on hypothalamus-pituitary-adrenal axis functioning and regulation as well as on behaviors linked to anxiety and depression traits in animals. We show that CBG deficiency in mice results in markedly reduced total circulating corticosterone at rest and in response to stress. Interestingly, free corticosterone concentrations are normal at rest but present a reduced surge after stress in transcortin-deficient mice. No differences were detected between transcortin-deficient mice for anxiety-related traits. However, transcortin-deficient mice display increased immobility in the forced-swimming test and markedly enhanced learned helplessness after prolonged uncontrollable stress. The latter is associated with an approximately 30% decrease in circulating levels of free corticosterone as well as reduced Egr-1 mRNA expression in hippocampus in CBG-deficient mice. Additionally, transcortin deficient mice show no sensitization to cocaine-induced locomotor responses, a well described corticosterone-dependent test. Thus, transcortin deficiency leads to insufficient glucocorticoid signaling and altered behavioral responses after stress. These findings uncover the critical role of plasma transcortin in providing an adequate endocrine and behavioral response to stress. PMID- 20022934 TI - Functional role of c-Jun-N-terminal kinase in feeding regulation. AB - c-Jun-N-terminal kinase (JNK) is a signaling molecule that is activated by proinflammatory signals, endoplasmic reticulum (ER) stress, and other environmental stressors. Although JNK has diverse effects on immunological responses and insulin resistance in peripheral tissues, a functional role for JNK in feeding regulation has not been established. In this study, we show that central inhibition of JNK activity potentiates the stimulatory effects of glucocorticoids on food intake and that this effect is abolished in mice whose agouti-related peptide (AgRP) neurons are degenerated. JNK1-deficient mice feed more upon central administration of glucocorticoids, and glucocorticoid receptor nuclear immunoreactivity is enhanced in the AgRP neurons. JNK inhibition in hypothalamic explants stimulates Agrp expression, and JNK1-deficient mice exhibit increased Agrp expression, heightened hyperphagia, and weight gain during refeeding. Our study shows that JNK1 is a novel regulator of feeding by antagonizing glucocorticoid function in AgRP neurons. Paradoxically, JNK1 mutant mice feed less and lose more weight upon central administration of insulin, suggesting that JNK1 antagonizes insulin function in the brain. Thus, JNK may integrate diverse metabolic signals and differentially regulate feeding under distinct physiological conditions. PMID- 20022936 TI - Breakthrough bacteraemia with a susceptible Enterococcus faecalis during tigecycline monotherapy. PMID- 20022935 TI - Stress process model for individuals with dementia. AB - PURPOSE: Individuals with dementia (IWDs) face particular challenges in managing and coping with their illness. The experience of dementia may be affected by the etiology, stage, and severity of symptoms, preexisting and related chronic conditions, and available informal and formal supportive services. Although several studies have examined particular features of IWD's illness experience, few draw upon a conceptual model that outlines the global illness experience and the resulting stressors that commence with symptom onset, proliferate over time, and continue through the later stages of cognitive loss. Building on the work of Pearlin and colleagues (1990, Caregiving and the stress process: An overview of concepts and their measures. The Gerontologist, 30, 583-594), this article proposes a stress process model (SPM) for IWDs that conceptualizes and examines the illness experience of IWDs. IMPLICATIONS: The proposed SPM for IWDs serves as a guide to (a) consider and understand the short- and long-term complexities of the illness experience for IWDs, (b) investigate specific hypotheses by outlining key stressors in the illness experience and by positing relationships among stressors and outcomes, and (c) help inform the development of interventions to prevent or reduce the negative stressors and enhance the positive experiences of living with a dementing illness. PMID- 20022937 TI - Molecular mechanisms of Bartonella henselae resistance to azithromycin, pradofloxacin and enrofloxacin. PMID- 20022938 TI - In vitro selection of Plasmodium falciparum drug-resistant parasite lines. AB - The in vitro selection of antimicrobial resistance in important pathogens can provide critical information on the genetic basis of drug resistance, and such information can be used to predict, anticipate and even contain the spread of resistance in clinical practice. For instance, the discovery of the role of pfmdr1 in mefloquine resistance in malaria parasites resulted from in vitro studies. However, the in vitro selection of resistance is difficult, challenging and time consuming. In this review, we discuss the key parameters that impact on the efficiency of the in vitro selection of resistance, and propose strategies to improve and streamline this process. PMID- 20022939 TI - Prosthetic hip joint infection with a Streptococcus agalactiae isolate not susceptible to penicillin G and ceftriaxone. PMID- 20022940 TI - Non-diabetic metabolic syndrome and obesity do not affect serum paraoxonase and arylesterase activities but do affect oxidative stress and inflammation. AB - OBJECTIVE: Paraoxonase-1 (PON-1), which has PON and arylesterase activities, is a high-density lipoprotein (HDL)-bound antioxidant enzyme that inhibits atherosclerosis. Diabetes has been shown to have an impact on oxidative stress. The effect of metabolic syndrome (MetS) on oxidative stress and PON-1 has been shown before, and PON-1 has been found to be related with accelerated atherogenesis. This study aimed to determine the oxidative state and PON and arylesterase activities in non-diabetic MetS and non-MetS obese patients. DESIGN: Thirty obese patients (3 M and 27 F) without MetS, 40 non-diabetic obese patients (3 M and 37 F) with MetS, and 30 controls (2 M and 28 F) were enrolled. METHODS: A 75 g glucose tolerance test was performed. PON-1, PON, arylesterase, total antioxidant status (TAS), high-sensitive C-reactive protein (hsCRP), and metabolic parameters were analyzed. RESULTS: PON and arylesterase activities were similar between the groups, while TAS was low in both MetS and obese groups compared to controls (P<0.01 and P<0.05 respectively). CRP was higher in the MetS group compared with the obese and control groups (P<0.01 and P<0.001 respectively). In both the obese and MetS groups, CRP showed a positive correlation with body mass index (BMI). TAS was negatively correlated with BMI, waist circumference, triglyceride levels, and systolic and diastolic blood pressures (P<0.001). CONCLUSIONS: Oxidative stress is altered in non-diabetic MetS and non-MetS obese patients, but PON and arylesterase activities seem not to be affected. This result may be due to the absence of diabetes, the most severe form of altered carbohydrate metabolism. PMID- 20022941 TI - Beta-cell development and turnover during prenatal life in humans. AB - INTRODUCTION: beta-cell regeneration is an area under active investigation for the future treatment of diabetes, but little is known about the patterns and dynamics of prenatal beta-cell development in humans. In particular, the quantitative changes in beta-cell mass in the developing pancreas have not been elucidated in detail. We addressed the following questions in prenatal humans: i) what is the timing of beta-cell occurrence and islet growth? ii) What are the dynamics of beta-cell replication and apoptosis? METHODS: Pancreatic tissue was obtained from 65 human embryos and foetuses aged between 8 weeks post conception (p.c.) and birth. Sections were stained for insulin, glucagon, Ki67 (proliferation marker), TUNEL (apoptosis marker) and CD31 (blood vessel marker), and morphometric analyses were performed. RESULTS: beta-cells were detected from gestational week 9 onward, whereas glucagon expression was detected already at week 8. The fractional beta-cell area of the pancreas increased in a linear fashion until birth (r=0.60, P<0.001). The first endocrine cells were found within or adjacent to the primitive ductal epithelium. beta-cell replication was readily detected in the newly forming islets already starting at week 9 p.c. (average frequency 2.8+/-0.4%). A small percentage of cells co-expressed insulin and glucagon during the early foetal period. There was a close relationship between the development of endocrine islets and blood vessels during all stages of prenatal pancreas development suggesting a possible interaction between both cell types. The frequency of beta-cell apoptosis was relatively high throughout all ages (1.5+/-0.3%). CONCLUSIONS: beta-cell differentiation in humans occurs from week 9 p.c. onward. The first endocrine cells are closely associated with the ductal epithelium suggesting differentiation from precursor cells. High rates of beta-cell replication suggest that this mechanism plays an important role in the prenatal expansion of beta-cell mass. PMID- 20022942 TI - An anticoagulant RNA aptamer that inhibits proteinase-cofactor interactions within prothrombinase. AB - The interaction of factor Xa with factor Va on membranes to form prothrombinase profoundly increases the rate of the proteolytic conversion of prothrombin to thrombin. We present the characterization of an RNA aptamer (RNA(11F7t)) selected from a combinatorial library based on its ability to bind factor Xa. We show that RNA(11F7t) inhibits thrombin formation catalyzed by prothrombinase without obscuring the active site of Xa within the enzyme complex. Selective inhibition of protein substrate cleavage arises from the ability of the aptamer to bind to factor Xa and exclude interactions between the proteinase and cofactor within prothrombinase. Competition for enzyme complex assembly results from the binding of RNA(11F7t) to factor Xa with nanomolar affinity in a Ca(2+)-dependent interaction. RNA(11F7t) binds equivalently to the zymogen factor X as well as derivatives lacking gamma-carboxyglutamic acid residues. We suggest that the ability of RNA(11F7t) to compete for the Xa-Va interaction with surprisingly high affinity likely reflects a significant contribution from its ability to indirectly impact regions of Xa that participate in the proteinase-cofactor interaction. Thus, despite the complexity of the macromolecular interactions that underlie the assembly of prothrombinase, efficient inhibition of enzyme complex assembly and thrombin formation can be achieved by tight binding ligands that target factor Xa in a discrete manner. PMID- 20022943 TI - Enhanced formation of disulfide-bridged dimer (Fab-PE38)2 utilizing repeats of the Fab binding domain of protein G. AB - Fab-PE38 used in this study is B3(Fab)-ext-PE38, and it is an antibody toxin that is made by fusing the Pseudomonas exotoxin to the Fab domain of B3 antibody. This antibody toxin selectively binds to cancer cells and kills the target cancer cells. B3(Fab)-ext-PE38 has a cysteine residue on the ext sequence, and (B3(Fab) ext-PE38)(2) is the disulfide-bridged dimer of the B3(Fab)-ext-PE38 monomer. (B3(Fab)-ext-PE38)(2) has been found to have 11-fold higher cytotoxicity on the CRL-1739 cell line than monomeric B3(scFv)-PE38. We made a recombinant tandem repeat of the domain III of Streptococcal protein G that has Fab binding property up to seven repeats. Multiple monomers were found to form non-covalent complexes with this tandem repeat. Complexes were purified by size-exclusion chromatography, and we could enhance the production of the disulfide-bridged dimer by reduction and oxidation of the complexes. The tandem repeat makes close intermolecular interactions between monomers possible, and the use of it greatly enhances the yield of the disulfide-bridged dimer. PMID- 20022944 TI - Her4 and Her2/neu tyrosine kinase domains dimerize and activate in a reconstituted in vitro system. AB - Her4 (ErbB-4) and Her2/neu (ErbB-2) are receptor-tyrosine kinases belonging to the epidermal growth factor receptor (EGFR) family. Crystal structures of EGFR and Her4 kinase domains demonstrate kinase dimerization and activation through an allosteric mechanism. The kinase domains form an asymmetric dimer, where the C lobe surface of one monomer contacts the N-lobe of the other monomer. EGFR kinase dimerization and activation in vitro was previously reported using a nickel chelating lipid-liposome system, and we now apply this system to all other members of the EGFR family. Polyhistidine-tagged Her4, Her2/neu, and Her3 kinase domains are bound to these nickel-liposomes and are brought to high local concentration, mimicking what happens to full-length receptors in vivo following ligand binding. Addition of nickel-liposomes to Her4 kinase domain results in 40 fold activation in kinase activity and marked enhancement of C-terminal tail autophosphorylation. Activation of Her4 shows a sigmoidal dependence on kinase concentration, consistent with a cooperative process requiring kinase dimerization. Her2/neu kinase activity is also activated by nickel-liposomes, and is increased further by heterodimerization with Her3 or Her4. The ability of Her3 and Her4 to heterodimerize and activate other family members is studied in vitro. Her3 kinase domain readily activates Her2/neu but is a poor activator of Her4, which differs from the prediction made by the asymmetric dimer model. Mutation of Her3 residues (952)ENI(954) to the corresponding sequence in Her4 enhanced the ability of Her3 to activate Her4, demonstrating that sequence differences on the C-lobe surface influence the heterodimerization and activation of ErbB kinase domains. PMID- 20022945 TI - Mg2+ dependence of 70 S ribosomal protein flexibility revealed by hydrogen/deuterium exchange and mass spectrometry. AB - The ribosome from Escherichia coli requires a specific concentration of Mg(2+) to maintain the 70 S complex formation and allow protein synthesis, and then the structure must be stable and flexible. How does the ribosome acquire these conflicting factors at the same time? Here, we investigated the hydrogen/deuterium exchange of 52 proteins in the 70 S ribosome, which controlled stability and flexibility under various Mg(2+) concentrations, using mass spectrometry. Many proteins exhibited a sigmoidal curve for Mg(2+) concentration dependence, incorporating more deuterium at lower Mg(2+) concentration. By comparing deuterium incorporation with assembly, we have discovered a typical mechanism of complexes for acquiring both stability and flexibility at the same time. In addition, we got information of the localization of flexibility in ribosomal function by the analysis of related proteins with stalk protein, tRNA, mRNA, and nascent peptide, and demonstrate the relationship between structure, assembly, flexibility, and function of the ribosome. PMID- 20022946 TI - mTOR Ser-2481 autophosphorylation monitors mTORC-specific catalytic activity and clarifies rapamycin mechanism of action. AB - The mammalian target of rapamycin (mTOR) Ser/Thr kinase signals in at least two multiprotein complexes distinguished by their different partners and sensitivities to rapamycin. Acute rapamycin inhibits signaling by mTOR complex 1 (mTORC1) but not mTOR complex 2 (mTORC2), which both promote cell growth, proliferation, and survival. Although mTORC2 regulation remains poorly defined, diverse cellular mitogens activate mTORC1 signaling in a manner that requires sufficient levels of amino acids and cellular energy. Before the identification of distinct mTOR complexes, mTOR was reported to autophosphorylate on Ser-2481 in vivo in a rapamycin- and amino acid-insensitive manner. These results suggested that modulation of mTOR intrinsic catalytic activity does not universally underlie mTOR regulation. Here we re-examine the regulation of mTOR Ser-2481 autophosphorylation (Ser(P)-2481) in vivo by studying mTORC-specific Ser(P)-2481 in mTORC1 and mTORC2, with a primary focus on mTORC1. In contrast to previous work, we find that acute rapamycin and amino acid withdrawal markedly attenuate mTORC1-associated mTOR Ser(P)-2481 in cycling cells. Although insulin stimulates both mTORC1- and mTORC2-associated mTOR Ser(P)-2481 in a phosphatidylinositol 3 kinase-dependent manner, rapamycin acutely inhibits insulin-stimulated mTOR Ser(P)-2481 in mTORC1 but not mTORC2. By interrogating diverse mTORC1 regulatory input, we find that without exception mTORC1-activating signals promote, whereas mTORC1-inhibitory signals decrease mTORC1-associated mTOR Ser(P)-2481. These data suggest that mTORC1- and likely mTORC2-associated mTOR Ser-2481 autophosphorylation directly monitors intrinsic mTORC-specific catalytic activity and reveal that rapamycin inhibits mTORC1 signaling in vivo by reducing mTORC1 catalytic activity. PMID- 20022947 TI - ERp57 modulates STAT3 signaling from the lumen of the endoplasmic reticulum. AB - ERp57 is an endoplasmic reticulum (ER) resident thiol disulfide oxidoreductase. Using the gene trap technique, we created a ERp57-deficient mouse model. Targeted deletion of the Pdia3 gene, which encodes ERp57, in mice is embryonic lethal at embryonic day (E) 13.5. Beta-galactosidase reporter gene analysis revealed that ERp57 is expressed early on during blastocyst formation with the highest expression in the inner cell mass. In early stages of mouse embryonic development (E11.5) there is a relatively low level of expression of ERp57. As the embryos developed, ERp57 became highly expressed in both the brain and the lungs (E15.5 and E18.5). The absence of ERp57 has no impact on ER morphology; expression of ER associated chaperones and folding enzymes, ER stress, or apoptosis. ERp57 has been reported to interact with STAT3 (signal transducer and activator of transcription)-DNA complexes. We show here that STAT3-dependent signaling is increased in the absence of ERp57 and this can be rescued by expression of ER targeted ERp57 but not by cytoplasmic-targeted protein, indicating that ERp57 affects STAT3 signaling from the lumen of the ER. ERp57 effects on STAT3 signaling are enhanced by ER luminal complex formation between ERp57 and calreticulin. In conclusion, we show that ERp57 deficiency in mouse is embryonic lethal at E13.5 and ERp57-dependent modulation of STAT3 signaling may contribute to this phenotype. PMID- 20022948 TI - TRPC3 regulates agonist-stimulated Ca2+ mobilization by mediating the interaction between type I inositol 1,4,5-trisphosphate receptor, RACK1, and Orai1. AB - There is a body of evidence suggesting that Ca(2+) handling proteins assemble into signaling complexes required for a fine regulation of Ca(2+) signals, events that regulate a variety of critical cellular processes. Canonical transient receptor potential (TRPC) and Orai proteins have both been proposed to form Ca(2+)-permeable channels mediating Ca(2+) entry upon agonist stimulation. A number of studies have demonstrated that inositol 1,4,5-trisphosphate receptors (IP(3)Rs) interact with plasma membrane TRPC channels; however, at present there is no evidence supporting the interaction between Orai proteins and IP(3)Rs. Here we report that treatment with thapsigargin or cellular agonists results in association of Orai1 with types I and II IP(3)Rs. In addition, we have found that TRPC3, RACK1 (receptor for activated protein kinase C-1), and STIM1 (stromal interaction molecule 1) interact with Orai1 upon stimulation with agonists. TRPC3 expression silencing prevented both the interaction of Orai1 with TRPC3 and, more interestingly, the association of Orai1 with the type I IP(3)R, but not with the type II IP(3)R, thus suggesting that TRPC3 selectively mediates interaction between Orai1 and type I IP(3)R. In addition, TRPC3 expression silencing attenuated ATP- and CCh-stimulated interaction between RACK1 and the type I IP(3)R, as well as Ca(2+) release and entry. In conclusion, our results indicate that agonist stimulation results in the formation of an Orai1-STIM1-TRPC3-RACK1 type I IP(3)R complex, where TRPC3 plays a central role. This Ca(2+) signaling complex might be important for both agonist-induced Ca(2+) release and entry. PMID- 20022949 TI - Stimulation of inducible nitric oxide by hepatitis B virus transactivator protein HBx requires MTA1 coregulator. AB - Nitric oxide has been implicated in the pathogenesis of inflammatory disorders, including hepatitis B virus-associated hepatocellular carcinoma. Transactivator protein HBx, a major regulator of cellular responses of hepatitis B virus, is known to induce the expression of MTA1 (metastasis-associated protein 1) coregulator via NF-kappaB signaling in hepatic cells. However, the underlying mechanism of HBx regulation of the inducible nitric-oxide synthase (iNOS) pathway remains unknown. Here we provide evidence that MTA1 is a positive regulator of iNOS transcription and plays a mechanistic role in HBx stimulation of iNOS expression and activity. We found that the HBx-MTA1 complex is recruited onto the human iNOS promoter in an NF-kappaB-dependent manner. Pharmacological inhibition of the NF-kappaB signaling prevented the ability of HBx to stimulate the transcription, the expression, and the activity of iNOS; nevertheless, these effects could be substantially rescued by MTA1 dysregulation. We further discovered that HBx-mediated stimulation of MTA1 is paralleled by the suppression of miR-661, a member of the small noncoding RNAs, recently shown to target MTA1. We observed that miR-661 controls of MTA1 expression contributed to the expression and activity of iNOS in HBx-expressing HepG2 cells. Accordingly, depletion of MTA1 by either miR-661 or siRNA in HBx-expressing cells severely impaired the ability of HBx to modulate the endogenous levels of iNOS and nitrite production. Together, these findings reveal an inherent role of MTA1 in HBx regulation of iNOS expression and consequently its function in the liver cancer cells. PMID- 20022950 TI - Dissecting the mechanism of insulin resistance using a novel heterodimerization strategy to activate Akt. AB - Insulin resistance can occur in response to many different external insults, including chronic exposure to insulin itself as well as other agonists such as dexamethasone. It is generally thought that such defects arise due to a defect(s) at an early stage in the insulin signaling cascade. One model suggests that this involves activation of the mammalian target of rapamycin/S6 kinase pathway, which inactivates insulin receptor substrate via Ser/Thr phosphorylation. However, we have recently shown that insulin receptor substrate is not a major node for insulin resistance defects. To explore the mechanism of insulin resistance, we have developed a novel system to activate Akt independently of its upstream effectors as well as other insulin-responsive pathways such as mitogen-activated protein kinase. 3T3-L1 adipocytes were rendered insulin-resistant either with chronic insulin or dexamethasone treatment, but conditional activation of Akt2 stimulated hemagglutinin-tagged glucose transporter 4 translocation to the same extent in these insulin-resistant and control cells. However, addition of insulin to cells in which Akt was conditionally activated resulted in a reversion to the insulin-resistant state, indicating a feedforward inhibitory mechanism activated by insulin itself. This effect was overcome with wortmannin, implicating a role for phosphatidylinositol 3-kinase in this inhibitory process. We conclude that in chronic insulin- and dexamethasone-treated cells, acute activation with insulin itself is required to activate a feedforward inhibitory pathway likely emanating from phosphatidylinositol 3-kinase that converges on a target downstream of Akt to cause insulin resistance. PMID- 20022951 TI - Minor modifications of the C-terminal helix reschedule the favored chemical reactions catalyzed by theta class glutathione transferase T1-1. AB - Adaptive responses to novel toxic challenges provide selective advantages to organisms in evolution. Glutathione transferases (GSTs) play a pivotal role in the cellular defense because they are main contributors to the inactivation of genotoxic compounds of exogenous as well as of endogenous origins. GSTs are promiscuous enzymes catalyzing a variety of chemical reactions with numerous alternative substrates. Despite broad substrate acceptance, individual GSTs display pronounced selectivities such that only a limited number of substrates are transformed with high catalytic efficiency. The present study shows that minor structural changes in the C-terminal helix of mouse GST T1-1 induce major changes in the substrate-activity profile of the enzyme to favor novel chemical reactions and to suppress other reactions catalyzed by the parental enzyme. PMID- 20022952 TI - Dissecting the factors involved in the locomotion mode of neuronal migration in the developing cerebral cortex. AB - Neuronal migration is essential for proper cortical layer formation and brain function, because migration defects result in neurological disorders such as mental retardation and epilepsy. Neuronal migration is divided into several contiguous steps: early phase (multipolar mode), locomotion mode, and terminal translocation mode. The locomotion mode covers most of the migration route and thereby is the main contributor to cortical layer formation. However, analysis of the molecular mechanisms regulating this mode is difficult due to the secondary effects of defects at the early phase of migration. In this study, we established an ex vivo chemical inhibitor screening, allowing us to directly analyze the locomotion mode of migration. Roscovitine and PP2, inhibitors for Cdk5 and Src family kinases, respectively, suppressed the locomotion mode of migration. In line with this, a small percentage of Cdk5- or Src family kinase (Fyn)-knockdown cells exhibited locomoting morphology but retarded migration, although the majority of cells were stalled at the early phase of migration. We also showed that rottlerin, widely used as a specific inhibitor for protein kinase Cdelta (PKCdelta), suppressed the locomotion mode. Unexpectedly, however, the dominant negative form as well as RNA interference for PKCdelta hardly affected the locomotion, whereas they may disturb terminal translocation. In addition, we found JNK to be a potential downstream target of rottlerin. Taken together, our novel chemical inhibitor screening provides evidence that Cdk5 and Src family kinases regulate the locomotion mode of neuronal migration. It also uncovered roles for Fyn and PKCdelta in the early and final phases of migration, respectively. PMID- 20022953 TI - The transcription activity of Gis1 is negatively modulated by proteasome-mediated limited proteolysis. AB - The transcriptional response to environmental changes has to be prompt but appropriate. Previously, it has been shown that the Gis1 transcription factor is responsible for regulating the expression of postdiauxic shift genes in response to nutrient starvation, and this transcription regulation is dependent upon the Rim15 kinase. Here we demonstrate that the activity of Gis1 is negatively modulated by proteasome-mediated limited proteolysis. Limited degradation of Gis1 by the proteasome leads to the production of smaller variants, which have weaker transcription activities than the full-length protein. The coiled-coil domain, absent from the smaller variants, is part of the second transcription activation domain in Gis1 and is essential for both the limited proteolysis of Gis1 and its full activity. Endogenous Gis1 and its variants, regardless of their transcription capabilities, activate transcription in a Rim15-dependent manner. However, when the full-length Gis1 accumulates in cells due to overexpression or inhibition of the proteasome function, transcription activation by Gis1 is no longer solely controlled by Rim15. Together, these data strongly indicate that the function of the limited degradation is to ensure that Gis1-dependent transcription is strictly regulated by the Rim15 kinase. Furthermore, we have revealed that the kinase activity of Rim15 is essential for this regulation. PMID- 20022954 TI - Vaccinia virus protein F1L is a caspase-9 inhibitor. AB - Apoptosis plays important roles in host defense, including the elimination of virus-infected cells. The executioners of apoptosis are caspase family proteases. We report that vaccinia virus-encoded F1L protein, previously recognized as anti apoptotic viral Bcl-2 family protein, is a caspase-9 inhibitor. F1L binds to and specifically inhibits caspase-9, the apical protease in the mitochondrial cell death pathway while failing to inhibit other caspases. In cells, F1L inhibits apoptosis and proteolytic processing of caspases induced by overexpression of caspase-9 but not caspase-8. An N-terminal region of F1L preceding the Bcl-2-like fold accounts for caspase-9 inhibition and significantly contributes to the anti apoptotic activity of F1L. Viral F1L thus provides the first example of caspase inhibition by a Bcl-2 family member; it functions both as a suppressor of proapoptotic Bcl-2 family proteins and as an inhibitor of caspase-9, thereby neutralizing two sequential steps in the mitochondrial cell death pathway. PMID- 20022955 TI - Identification of a novel inhibitor of coactivator-associated arginine methyltransferase 1 (CARM1)-mediated methylation of histone H3 Arg-17. AB - Methylation of the arginine residues of histones by methyltransferases has important consequences for chromatin structure and gene regulation; however, the molecular mechanism(s) of methyltransferase regulation is still unclear, as is the biological significance of methylation at particular arginine residues. Here, we report a novel specific inhibitor of coactivator-associated arginine methyltransferase 1 (CARM1; also known as PRMT4) that selectively inhibits methylation at arginine 17 of histone H3 (H3R17). Remarkably, this plant-derived inhibitor, called TBBD (ellagic acid), binds to the substrate (histone) preferentially at the signature motif, "KAPRK," where the proline residue (Pro 16) plays a critical role for interaction and subsequent enzyme inhibition. In a promoter-specific context, inhibition of H3R17 methylation represses expression of p21, a p53-responsive gene, thus implicating a possible role for H3 Arg-17 methylation in tumor suppressor function. These data establish TBBD as a novel specific inhibitor of arginine methylation and demonstrate substrate sequence directed inhibition of enzyme activity by a small molecule and its physiological consequence. PMID- 20022956 TI - Isolation and characterization of cytoplasmic cofilin-actin rods. AB - Cofilin-actin bundles (rods), which form in axons and dendrites of stressed neurons, lead to synaptic dysfunction and may mediate cognitive deficits in dementias. Rods form abundantly in the cytoplasm of non-neuronal cells in response to many treatments that induce rods in neurons. Rods in cell lysates are not stable in detergents or with added calcium. Rods induced by ATP-depletion and released from cells by mechanical lysis were first isolated from two cell lines expressing chimeric actin-depolymerizing factor (ADF)/cofilin fluorescent proteins by differential and equilibrium sedimentation on OptiPrep gradients and then from neuronal and non-neuronal cells expressing only endogenous proteins. Rods contain ADF/cofilin and actin in a 1:1 ratio. Isolated rods are stable in dithiothreitol, EGTA, Ca(2+), and ATP. Cofilin-GFP-containing rods are stable in 500 mM NaCl, whereas rods formed from endogenous proteins are significantly less stable in high salt. Proteomic analysis of rods formed from endogenous proteins identified other potential components whose presence in rods was examined by immunofluorescence staining of cells. Only actin and ADF/cofilin are in rods during all phases of their formation; furthermore, the rapid assembly of rods in vitro from these purified proteins at physiological concentration shows that they are the only proteins necessary for rod formation. Cytoplasmic rod formation is inhibited by cytochalasin D and jasplakinolide. Time lapse imaging of rod formation shows abundant small needle-shaped rods that coalesce over time. Rod filament lengths measured by ultrastructural tomography ranged from 22 to 1480 nm. These results suggest rods form by assembly of cofilin-actin subunits, followed by self-association of ADF/cofilin-saturated F-actin. PMID- 20022957 TI - AAA+ chaperone ClpX regulates dynamics of prokaryotic cytoskeletal protein FtsZ. AB - AAA(+) chaperone ClpX has been suggested to be a modulator of prokaryotic cytoskeletal protein FtsZ, but the details of recognition and remodeling of FtsZ by ClpX are largely unknown. In this study, we have extensively investigated the nature of FtsZ polymers and mechanisms of ClpX-regulated FtsZ polymer dynamics. We found that FtsZ polymerization is inhibited by ClpX in an ATP-independent manner and that the N-terminal domain of ClpX plays a crucial role for the inhibition of FtsZ polymerization. Single molecule analysis with high speed atomic force microscopy directly revealed that FtsZ polymer is in a dynamic equilibrium between polymerization and depolymerization on a time scale of several seconds. ClpX disassembles FtsZ polymers presumably by blocking reassembly of FtsZ. Furthermore, Escherichia coli cells overproducing ClpX and N terminal domain of ClpX show filamentous morphology with abnormal localization of FtsZ. These data together suggest that ClpX modulates FtsZ polymer dynamics in an ATP-independent fashion, which is achieved by interaction between the N-terminal domain of ClpX and FtsZ monomers or oligomers. PMID- 20022958 TI - Deficiency in APOBEC2 leads to a shift in muscle fiber type, diminished body mass, and myopathy. AB - The apoB RNA-editing enzyme, catalytic polypeptide-like (APOBEC) family of proteins includes APOBEC1, APOBEC3, and activation-induced deaminase, all of which are zinc-dependent cytidine deaminases active on polynucleotides and involved in RNA editing or DNA mutation. In contrast, the biochemical and physiological functions of APOBEC2, a muscle-specific member of the family, are unknown, although it has been speculated, like APOBEC1, to be an RNA-editing enzyme. Here, we show that, although expressed widely in striated muscle (with levels peaking late during myoblast differentiation), APOBEC2 is preferentially associated with slow-twitch muscle, with its abundance being considerably greater in soleus compared with gastrocnemius muscle and, within soleus muscle, in slow as opposed to fast muscle fibers. Its abundance also decreases following muscle denervation. We further show that APOBEC2-deficient mice harbor a markedly increased ratio of slow to fast fibers in soleus muscle and exhibit an approximately 15-20% reduction in body mass from birth onwards, with elderly mutant animals revealing clear histological evidence of a mild myopathy. Thus, APOBEC2 is essential for normal muscle development and maintenance of fiber-type ratios; although its molecular function remains to be identified, biochemical analyses do not especially argue for any role in RNA editing. PMID- 20022959 TI - Cell signaling, internalization, and nuclear localization of the angiotensin converting enzyme in smooth muscle and endothelial cells. AB - The angiotensin converting enzyme (ACE) catalyzes the extracellular formation of angiotensin II, and degradation of bradykinin, thus regulating blood pressure and renal handling of electrolytes. We have previously shown that exogenously added ACE elicited transcriptional regulation independent of its enzymatic activity. Because transcriptional regulation generates from protein-DNA interactions within the cell nucleus we have investigated the initial cellular response to exogenous ACE and the putative internalization of the enzyme in smooth muscle cells (SMC) and endothelial cells (EC). The following phenomena were observed when ACE was added to cells in culture: 1) it bound to SMC and EC with high affinity (K(d) = 361.5 +/- 60.5 pM) and with a low binding occupancy (B(max) = 335.0 +/- 14.0 molecules/cell); 2) it triggered cellular signaling resulting in late activation of focal adhesion kinase and SHP2; 3) it modulated platelet-derived growth factor receptor-beta signaling; 4) it was endocytosed by SMC and EC; and 5) it transited through the early endosome, partially occupied the late endosome and the lysosome, and was localized to the nuclei. The incorporation of ACE or a fragment of it into the nuclei reached saturation at 120 min, and was preceded by a lag time of 40 min. Internalized ACE was partially cleaved into small fragments. These results revealed that extracellular ACE modulated cell signaling properties, and that SMC and EC have a pathway for delivery of extracellular ACE to the nucleus, most likely involving cell surface receptor(s) and requiring transit through late endosome/lysosome compartments. PMID- 20022960 TI - Heparan sulfate is required for embryonic stem cells to exit from self-renewal. AB - Pluripotent embryonic stem cells (ESCs) must select between alternative fates of self-renewal and lineage commitment at each division during continuous proliferation. Heparan sulfate (HS) is a highly sulfated polysaccharide and is present abundantly on the ESC surface. In this study, we investigated the role of HS in ESC self-renewal by examining Ext1(-/-) ESCs that are deficient in HS. We found that Ext1(-/-) ESCs retained their self-renewal potential but failed to transit from self-renewal to differentiation upon removal of leukemia inhibitory factor. Furthermore, we found that the aberrant cell fate commitment is caused by defects in fibroblast growth factor signaling, which directly retained high expression of the pluripotency gene Nanog in Ext1(-/-) ESCs. Therefore, our studies identified and defined HS as a novel factor that controls ESC fate commitment and also delineates that HS facilitates fibroblast growth factor signaling, which, in turn, inhibits Nanog expression and commits ESCs to lineage differentiation. PMID- 20022961 TI - Investigation of human telomerase holoenzyme assembly, activity, and processivity using disease-linked subunit variants. AB - After the initial discovery of human telomerase deficiency in the X-linked form of the bone marrow failure syndrome dyskeratosis congenita, mutations in genes encoding telomerase subunits have been identified in patients with a wide spectrum of disorders. Structure/function studies of disease-linked variants of human telomerase RNA (hTR) or telomerase reverse transcriptase (TERT) have exploited in vitro reconstitution of the enzyme catalytic core and/or a PCR amplified activity assay readout that would not reflect alterations of cellular RNP assembly efficiency, telomeric primer recognition, and/or repeat addition processivity. Here we used telomerase reconstitution in vivo and direct telomeric repeat primer extension activity assays to compare the ribonucleoprotein (RNP) assembly and activity properties of disease-linked subunit variants in holoenzyme context. Analysis of a large panel of hTR variants revealed numerous biochemical mechanisms for telomerase loss of function, including reduced association of hTR with TERT, reduced RNP catalytic activity, or loss in fidelity of telomeric repeat synthesis. An absolute correlation exists between hTR loss of function and hematopoietic deficiency, but there is no readily apparent telomerase deficiency imposed by an hTR variant linked to pulmonary fibrosis. Some disease-linked TERT variants have altered properties of holoenzyme assembly or repeat addition processivity, but other TERT variants linked to either pulmonary fibrosis or hematopoietic deficiency retained normal hTR interaction and RNP catalytic activity. Combined with additional hTR structure/function studies, our results establish a new resolution of insight into hTR structural requirements for hTR TERT interaction and for the catalytic cycle of human telomerase holoenzyme. PMID- 20022962 TI - A DDX5 S480A polymorphism is associated with increased transcription of fibrogenic genes in hepatic stellate cells. AB - We recently identified a missense single nucleotide polymorphism (SNP) in DDX5 (rs1140409, p.S480A) that enhances the risk of developing cirrhosis. DDX5 is an ATP-dependent RNA helicase and transcriptional modulator. We hypothesized that the activity of DDX5 in regulating fibrogenic gene transcription in hepatic stellate cells (HSCs) is altered by the S480A SNP. To test this, we employed two approaches: 1) transient overexpression of DDX5 cDNA or siRNA knockdown of endogenous DDX5, with replacement by either DDX5 wild type (WT) or SNP cDNA, or 2) stable expression of exogenous DDX5 WT and SNP in HSC lines. WT DDX5 mRNA in HSCs was inversely correlated with gene expression for alpha2(I) collagen, tissue inhibitor of metalloproteinase-1, and transforming growth factor-beta1. Stable DDX5 SNP-expressing cells had higher basal and transforming growth factor-beta1 stimulated expression and enhanced promoter activities of fibrogenic genes. DDX5 variant-expressing cells also had higher Smad3 and AP-1-responsive reporter activities. In a one-hybrid GAL4 system, co-expression of the DDX5 SNP variant with chimeras of GAL4 DNA binding domain linked to JunD or Sp1 displayed higher transactivation of a GAL4-responsive reporter than that of DDX5 WT. Increased fibrogenic gene expression in DDX5 SNP-expressing cells was associated with reduced recruitment of DDX5 homodimers to responsive promoters, but there was no difference in the recruitment of the co-repressor HDAC1 (histone deacetylase 1). These data suggest that DDX5 is a repressor of fibrogenic genes in HSCs through interaction with transcriptional complexes. The enhanced fibrogenic activity of the DDX5 risk variant is linked to a reduced repressive function toward these target genes. PMID- 20022963 TI - Inositol 1,4,5-trisphosphate 3-kinase-A is a new cell motility-promoting protein that increases the metastatic potential of tumor cells by two functional activities. AB - Cellular migration is an essential prerequisite for metastatic dissemination of cancer cells. This study demonstrates that the neuron/testis-specific F-actin targeted inositol 1,4,5-trisphosphate 3-kinase-A (ITPKA) is ectopically expressed in different human tumor cell lines and during tumor progression in the metastatic tumor model Balb-neuT. High expression of ITPKA increases invasive migration in vitro and metastasis in a xenograft SCID mouse model. Mechanistic studies show that ITPKA promotes migration of tumor cells by two different mechanisms as follows: growth factor independently high levels of ITPKA induce the formation of large cellular protrusions by directly modulating the actin cytoskeleton. The F-actin binding activity of ITPKA stabilizes and bundles actin filaments and thus increases the levels of cellular F-actin. In growth factor stimulated cells, the catalytically active domain enhances basal ITPKA-induced migration by activating store-operated calcium entry through production of inositol 1,3,4,5-tetrakisphosphate and subsequent inhibition of inositol phosphate 5-phosphatase. These two functional activities of ITPKA stimulating tumor cell migration place the enzyme among the potential targets of anti metastatic therapy. PMID- 20022964 TI - PemK toxin of Bacillus anthracis is a ribonuclease: an insight into its active site, structure, and function. AB - Bacillus anthracis genome harbors a toxin-antitoxin (TA) module encoding pemI (antitoxin) and pemK (toxin). This study describes the rPemK as a potent ribonuclease with a preference for pyrimidines (C/U), which is consistent with our previous study that demonstrated it as a translational attenuator. The in silico structural modeling of the PemK in conjunction with the site-directed mutagenesis confirmed the role of His-59 and Glu-78 as an acid-base couple in mediating the ribonuclease activity. The rPemK is shown to form a complex with the rPemI, which is in line with its function as a TA module. This rPemI-rPemK complex becomes catalytically inactive when both the proteins interact in a molar stoichiometry of 1. The rPemI displays vulnerability to proteolysis but attains conformational stability only upon rPemK interaction. The pemI-pemK transcript is shown to be up-regulated upon stress induction with a concomitant increase in the amount of PemK and a decline in the PemI levels, establishing the role of these modules in stress. The artificial perturbation of TA interaction could unleash the toxin, executing bacterial cell death. Toward this end, synthetic peptides are designed to disrupt the TA interaction. The peptides are shown to be effective in abrogating TA interaction in micromolar range in vitro. This approach can be harnessed as a potential antibacterial strategy against anthrax in the future. PMID- 20022965 TI - Regulation of endoplasmic reticulum stress-induced cell death by ATF4 in neuroectodermal tumor cells. AB - The neuroectodermal tumors neuroblastoma and melanoma represent biologically aggressive and chemoresistant cancers. The chemotherapeutic agents fenretinide and bortezomib induce apoptosis through endoplasmic reticulum (ER) stress in these tumor types. The aim of this study was to test the hypothesis that the early events of ER stress signaling and response pathways induced by fenretinide and bortezomib are mediated by the eukaryotic initiation factor 2alpha (eIF2alpha)-ATF4 signaling pathway. Treatment of neuroblastoma and melanoma cell lines with fenretinide, bortezomib, or thapsigargin resulted in induction of eIF2alpha signaling, characterized by increased expression of phosphorylated eIF2alpha, ATF4, ATF3, and GADD34. These events correlated with induction of the pro-apoptotic protein Noxa. The cytotoxic response, characterized by up regulation of Noxa and cell death, was dependent on ATF4, but not the ER-related pro-death signaling pathways involving GADD153 or IRE1. Although PERK-dependent phosphorylation of eIF2alpha enhanced ATF4 protein levels during ER stress, cell death in response to fenretinide, bortezomib, or thapsigargin was not abrogated by inhibition of eIF2alpha phosphorylation through PERK knockdown or overexpression of wild-type eIF2alpha. Furthermore, ATF4 induction in response to ER stress was dependent primarily on transcriptional activation, which occurred in a PERK- and phosphorylated eIF2alpha-independent manner. These results demonstrate that ATF4 mediates ER stress-induced cell death of neuroectodermal tumor cells in response to fenretinide or bortezomib. Understanding the complex regulation of cell death pathways in response to ER stress-inducing drugs has the potential to reveal novel therapeutic targets, thus allowing the development of improved treatment strategies to overcome chemoresistance. PMID- 20022966 TI - Nuclear localization of the p75 neurotrophin receptor intracellular domain. AB - The p75 neurotrophin receptor, a member of the tumor necrosis factor superfamily of receptors, undergoes an alpha-secretase-mediated release of its extracellular domain, followed by a gamma-secretase-mediated intramembrane cleavage. Like amyloid precursor protein and Notch, gamma-secretase cleavage of the p75 receptor releases an intracellular domain (ICD). However, it has been experimentally challenging to determine the precise subcellular localization and functional consequences of the p75 ICD. Here, we utilized a nuclear translocation assay and biochemical fractionation approaches to follow the fate of the ICD. We found that the p75 ICD can translocate to the nucleus to activate a green fluorescent protein reporter gene. Furthermore, the p75 ICD was localized in nuclear fractions. Chromatin immunoprecipitation experiments indicated that nerve growth factor induced the association of endogenous p75 with the cyclin E(1) promoter. Expression of the p75 ICD resulted in modulation of gene expression from this locus. These results suggest that the p75 ICD generated by gamma-secretase cleavage is capable of modulating transcriptional events in the nucleus. PMID- 20022967 TI - Outcomes of patients ventilated with synchronized intermittent mandatory ventilation with pressure support: a comparative propensity score study. AB - BACKGROUND: Few data are available regarding the benefits of one mode over another for ventilatory support. We set out to compare clinical outcomes of patients receiving synchronized intermittent mandatory ventilation with pressure support (SIMV-PS) compared with assist-control (A/C) ventilation as their primary mode of ventilatory support. METHODS: This was a secondary analysis of an observational study conducted in 349 ICUs from 23 countries. A propensity score stratified analysis was used to compare 350 patients ventilated with SIMV-PS with 1,228 patients ventilated with A/C ventilation. The primary outcome was in hospital mortality. RESULTS: In a logistic regression model, patients were more likely to receive SIMV-PS if they were from North America, had lower severity of illness, or were ventilated postoperatively or for trauma. SIMV-PS was less likely to be selected if patients were ventilated because of asthma or coma, or if they developed complications such as sepsis or cardiovascular failure during mechanical ventilation. In the stratified analysis according to propensity score, we did not find significant differences in the in-hospital mortality. After adjustment for propensity score, overall effect of SIMV-PS on in-hospital mortality was not significant (odds ratio, 1.04; 95% CI, 0.77-1.42; P = .78). CONCLUSIONS: In our cohort of ventilated patients, ventilation with SIMV-PS compared with A/C did not offer any advantage in terms of clinical outcomes, despite treatment-allocation bias that would have favored SIMV-PS. PMID- 20022968 TI - Evidence-based comparison of commercial interferon-gamma release assays for detecting active TB: a metaanalysis. AB - Test accuracy of interferon-gamma release assays (IGRAs) for diagnosing TB differs when using older or precommercial tools and inconsistent diagnostic criteria. This metaanalysis critically appraises studies investigating sensitivity and specificity of the commercial T-Spot.TB and the QuantiFERON-TB Gold In-Tube Assay (QFT-IT) among definitely confirmed TB cases. We searched Medline, EMBASE, and Cochrane bibliographies of relevant articles. Sensitivities, specificities, and indeterminate rates were pooled using a fixed effect model. Sensitivity of the tuberculin skin test (TST) was evaluated in the context of IGRA studies. In addition, the rates of indeterminates of both IGRAs were assessed. The pooled sensitivity of TST was 70% (95% CI, 0.67-0.72) compared with 81% (95% CI, 0.78-0.83) for the QFT-IT and 88% (95% CI, 0.85-0.90) for the T Spot.TB. Sensitivity increased to 84% (95%CI, 0.81-0.87) and 89% (95% CI, 0.86 0.91) for the QFT-IT and T-Spot.TB, respectively, when restricted to performance in developed countries. In contrast, specificity of the QFT-IT was 99% (95% CI, 0.98-1.00) vs 86% for the T-Spot.TB (95% CI, 0.81-0.90). The pooled rate of indeterminate results was low, 2.1% (95% CI, 0.02-0.023) for the QFT-IT and 3.8% (95% CI, 0.035-0.042) for the T-Spot.TB, increasing to 4.4% (95% CI, 0.039-0.05) and 6.1% (95% CI, 0.052-0.071), respectively, among immunosuppressed hosts. The newest commercial IGRAs are superior, in comparison with the TST, for detecting confirmed active TB disease, especially when performed in developed countries. PMID- 20022969 TI - Clinical management of pandemic 2009 influenza A(H1N1) infection. AB - Antiviral therapy and vaccination are important strategies for controlling pandemic 2009 influenza A(H1N1) but efficacy depends on the timing of administration and is often limited by supply shortage. Patients with dyspnea, tachypnea, evidence of hypoxemia, and pulmonary infiltrates on chest radiograph should be hospitalized. Patients with severe illness or underlying medical conditions that increase the risk of more severe disease should be treated with oseltamivir or zanamivir as soon as possible, without waiting for the results of laboratory tests. Lung-protective ventilation strategy with a low tidal volume and adequate pressure, in addition to a conservative fluid management approach, is recommended when treating adult patients with ARDS. Extracorporeal membrane oxygenation has emerged as an important rescue therapy for critically ill patients. Use of systemic steroids was associated with delayed viral clearance in severe acute respiratory syndrome and H3N2 infection. Low-dose corticosteroids may be considered in the treatment of refractory septic shock. Passive immunotherapy in the form of convalescent plasma or hyperimmune globulin may be explored as rescue therapy. More data are needed to explore the potential role of IV gamma globulin and other drugs with immunomodulating properties, such as statins, gemfibrozil, and N-acetyl-cysteine. Health-care workers must apply strict standard and droplet precautions when dealing with suspected and confirmed case and upgrade to airborne precautions when performing aerosol-generating procedures. Nonpharmacologic measures, such as early case isolation, household quarantine, school/workplace closure, good community hygiene, and restrictions on travel are useful measures in controlling an influenza pandemic at its early phase. PMID- 20022970 TI - Increased risk of myocardial infarction and stroke following exacerbation of COPD. AB - OBJECTIVE: Patients with COPD are at risk for cardiovascular events. This is attributed to increased systemic inflammation. The course of COPD is punctuated by exacerbations, which further increase systemic inflammation, but the risk of vascular events in the postexacerbation period has never been defined. METHODS: We analyzed data from 25,857 patients with COPD entered in The Health Improvement Network database over a 2-year period. Exacerbations were defined using a health care use definition of prescription of oral corticosteroids > 20 mg/d and/or selected oral antibiotics. The risk of myocardial infarction (MI) and stroke in the postexacerbation period was calculated relative to the patient's baseline risk using the self-controlled case series approach. RESULTS: We identified 524 MIs in 426 patients and 633 ischemic strokes in 482 patients. The incidence rates of MI and stroke were 1.1 and 1.4 per 100 patient-years, respectively. There was a 2.27-fold (95% CI, 1.1-4.7; P = .03) increased risk of MI 1 to 5 days after exacerbation (defined by prescription of both steroids and antibiotics). This relative risk diminished progressively with time and was not significantly different from the baseline MI risk at any other postexacerbation time interval. One in 2,513 exacerbations was associated with MI within 1 to 5 days. There was a 1.26-fold (95% CI, 1.0-1.6; P = .05) increased risk of stroke 1 to 49 days after exacerbation. CONCLUSION: The results suggest that exacerbations of COPD increase the risk of MI and stroke. This may have implications for therapy in both stable and exacerbated COPD. PMID- 20022971 TI - Adipokines in children with obstructive sleep apnea and the effects of treatment. AB - BACKGROUND: The objective of this study was to evaluate circulating adipokines concentration in children with and without obstructive sleep apnea (OSA) and to determine the effects of treatment of OSA on their plasma concentration. METHODS: Children with habitual snoring and symptoms suggestive of OSA were consecutively recruited. Their parents completed a sleep apnea symptom questionnaire, and the subjects underwent physical examination and an overnight polysomnography (PSG). OSA was diagnosed if they had an obstructive apnea index > 1. Fasting serum adiponectin, leptin, and lipid profiles were taken after overnight PSG. The subjects were divided into groups as obese, nonobese, and with and without OSA for comparison. RESULTS: One hundred forty-one children, of whom 96 were boys, with a median (interquartile range) age of 10.8 (8.5-12.8) years were recruited. Forty-three subjects had OSA. Subjects with OSA did not have significantly different adiponectin and leptin concentrations than those without OSA for both the obese and nonobese groups. Stepwise multiple linear regressions revealed that systolic BP, age, high-density lipoprotein cholesterol, and BMI z-score were independently associated with adiponectin, whereas diastolic BP, triglyceride, height, and BMI z-score were independently associated with leptin concentration. Sixteen children with OSA underwent treatment, and there was reduction in their plasma adiponectin concentration after intervention, but such change became insignificant after controlling for change in the BMI z-score. CONCLUSIONS: BMI rather than OSA was the main determinant of adipokines in children. PMID- 20022972 TI - Comparison of plethysmographic and helium dilution lung volumes: which is best for COPD? AB - BACKGROUND: Theoretical considerations and limited scientific evidence suggest that whole-body plethysmography overestimates lung volume in patients with severe airflow obstruction. We sought to compare plethysmography (Pleth)-, helium dilution (He)- and CT scan-derived lung volume measurements in a sample containing many patients with severe airflow obstruction. METHODS: We measured total lung capacity (TLC) in 132 patients at three hospitals, with monitored application of recommended techniques for Pleth and He measurements of lung volume and by thoracic CT scans obtained during breath hold at full inspiration. RESULTS: Average TLC among 132 subjects was 6.18 L (+/- 1.69 L) by Pleth-derived TLC, 5.55 L (+/- 1.39 L) by He-derived TLC, and 5.31 L (+/- 1.47) by CT scan derived TLC. Pleth-derived TLC was significantly greater than either He-derived TLC or CT scan-derived TLC (P < or = .001), whereas there was no significant difference between He-derived and CT scan-derived values. When examined separately, there were significant within-subject differences in TLC by measurement technique among subjects with airflow obstruction, but not among those without airflow obstruction. Plethysmographic overestimation of TLC was greatest among subjects with FEV(1) < 30% of predicted. CONCLUSIONS: In the setting of airflow obstruction, Pleth systematically overestimates lung volume relative to He or thoracic imaging despite adherence to current recommendations for proper measurement technique. PMID- 20022973 TI - Copy number variant detection in inbred strains from short read sequence data. AB - SUMMARY: We have developed an algorithm to detect copy number variants (CNVs) in homozygous organisms, such as inbred laboratory strains of mice, from short read sequence data. Our novel approach exploits the fact that inbred mice are homozygous at virtually every position in the genome to detect CNVs using a hidden Markov model (HMM). This HMM uses both the density of sequence reads mapped to the genome, and the rate of apparent heterozygous single nucleotide polymorphisms, to determine genomic copy number. We tested our algorithm on short read sequence data generated from re-sequencing chromosome 17 of the mouse strains A/J and CAST/EiJ with the Illumina platform. In total, we identified 118 copy number variants (43 for A/J and 75 for CAST/EiJ). We investigated the performance of our algorithm through comparison to CNVs previously identified by array-comparative genomic hybridization (array CGH). We performed quantitative PCR validation on a subset of the calls that differed from the array CGH data sets. PMID- 20022974 TI - webMGR: an online tool for the multiple genome rearrangement problem. AB - SUMMARY: The algorithm MGR enables the reconstruction of rearrangement phylogenies based on gene or synteny block order in multiple genomes. Although MGR has been successfully applied to study the evolution of different sets of species, its utilization has been hampered by the prohibitive running time for some applications. In the current work, we have designed new heuristics that significantly speed up the tool without compromising its accuracy. Moreover, we have developed a web server (webMGR) that includes elaborate web output to facilitate navigation through the results. AVAILABILITY: webMGR can be accessed via http://www.gis.a-star.edu.sg/~bourque. The source code of the improved standalone version of MGR is also freely available from the web site. SUPPLEMENTARY INFORMATION: Supplementary data are available at Bioinformatics online. PMID- 20022975 TI - RNA-Seq gene expression estimation with read mapping uncertainty. AB - MOTIVATION: RNA-Seq is a promising new technology for accurately measuring gene expression levels. Expression estimation with RNA-Seq requires the mapping of relatively short sequencing reads to a reference genome or transcript set. Because reads are generally shorter than transcripts from which they are derived, a single read may map to multiple genes and isoforms, complicating expression analyses. Previous computational methods either discard reads that map to multiple locations or allocate them to genes heuristically. RESULTS: We present a generative statistical model and associated inference methods that handle read mapping uncertainty in a principled manner. Through simulations parameterized by real RNA-Seq data, we show that our method is more accurate than previous methods. Our improved accuracy is the result of handling read mapping uncertainty with a statistical model and the estimation of gene expression levels as the sum of isoform expression levels. Unlike previous methods, our method is capable of modeling non-uniform read distributions. Simulations with our method indicate that a read length of 20-25 bases is optimal for gene-level expression estimation from mouse and maize RNA-Seq data when sequencing throughput is fixed. PMID- 20022976 TI - Identification of growth insensitive to ABA3 (gia3), a recessive mutation affecting ABA Signaling for the control of early post-germination growth in Arabidopsis thaliana. AB - The stress phytohormone ABA inhibits the developmental transition taking the mature embryo in the dry seed towards a young seedling. ABA also induces the accumulation of the basic leucine zipper (bZIP) transcription factor ABA insensitive 5 (ABI5) which, apart from blocking endosperm rupture, also protects the embryo by stimulating the expression of late embryogenesis abundant (LEA) genes that conferred osmotolerance during seed maturation. It is unknown whether ABA recruits additional embryonic pathways to control early seedling growth and fitness. Here we identify gia3 (growth insensitive to ABA3), a recessive locus in Arabidopsis mediating cotyledon cellular maturation and ABA-dependent repression of cotyledon expansion and greening. Microarray studies showed that expression of the essential mid-embryogenesis gene Maternal Embryo Effect 26 (MEE26) is induced by ABA during early seedling growth in wild-type (WT) or abi5 plants but not in gia3 mutants. However, we also show that the GIA3 locus controls ABA-dependent gene expression responses that partially overlap with those controlled by ABI5. Thus, the gia3 locus identifies an additional arm of ABA signaling, distinct from that controlled by ABI5, which recruits MEE26 expression and maintains cotyledon embryonic identity. Fine mapping localized the gia3 locus within a 1 Mb interval of chromosome 3, containing a large DNA insertion of a duplicated region of chromosome 2. It remains unknown at present whether gia3 phenotypes are the result of single or multiple genetic alterations. PMID- 20022977 TI - Selective excitation of photosystems in chloroplasts inside plant leaves observed by near-infrared laser-based fluorescence spectral microscopy. AB - In this study, we produced selective images of photosystems in plant chloroplasts in situ. We used a spectroimaging microscope, equipped with a near-infrared (NIR) laser that provided light at wavelengths mainly between 800 and 830 nm, to analyze chlorophyll autofluorescence spectra and images from chloroplasts in leaves of Zea mays at room temperature. Femtosecond laser excitation of chloroplasts in mesophyll cells revealed a spectral shape that was attributable to PSII and its antenna in the centers of grana spots. We found that a continuous wave emitted by the NIR laser at a wavelength as long as 820 nm induced chlorophyll autofluorescence with a high contribution from PSI through a one photon absorption mechanism. A spectral shape attributable to PSI and its antenna was thus obtained using continuous wave laser excitation of chloroplasts in bundle sheath cells. These highly pure spectra of photosystems were utilized for spectral decomposition at every intrachloroplast space to show distributions of PSI and PSII and their associated antenna. A new methodology using an NIR laser to detect the PSI/PSII ratio in single chloroplasts in leaves at room temperature is described. PMID- 20022978 TI - Alternative expression of vacuolar iron transporter and ferritin genes leads to blue/purple coloration of flowers in tulip cv. 'Murasakizuisho'. AB - Flowers of tulip cv. 'Murasakizuisho' have a purple perianth except for the bottom region, which is blue in color even though it has the same anthocyanin, delphinidin 3-O-rutinoside, as the entire perianth. The development of the blue coloration in the perianth bottom is due to complexation by anthocyanin, flavonol and iron (Fe), as well as a vacuolar iron transporter, TgVit1. Although transient expression of TgVit1 in the purple cells led to a color change to light blue, the coloration of the transformed cells did not coincide with the dark blue color of the cells of the perianth bottom. We thought that another factor is required for the blue coloration of the cells of perianth bottom. To examine the effect of ferritin (FER), an Fe storage protein, on blue color development, we cloned an FER gene (TgFER1) and performed expression analyses. TgFER1 transcripts were found in the cells located in the upper region of the petals along with purple color development by anthocyanin and were not found in the blue cells of the perianth bottom. This gene expression is in contrast to that of TgVit1, expressed only in the cells of the perianth bottom. Co-expression of TgVIT1 and TgFER-RNAi, constructed for suppressing endogenous TgFER1 by RNA interference (RNAi), changed the purple petal cells to a dark blue color similar to that of the natural perianth bottom. These results strongly suggest that TgVit1 expression and TgFER1 suppression are critical for the development of blue color in the perianth bottom. PMID- 20022979 TI - Critical communication. PMID- 20022981 TI - Selected reaction monitoring-mass spectrometric immunoassay responsive to parathyroid hormone and related variants. AB - BACKGROUND: Parathyroid hormone (PTH) assays able to distinguish between full length PTH (PTH1-84) and N-terminally truncated PTH (PTH7-84) are of increasing significance in the accurate diagnosis of endocrine and osteological diseases. We describe the discovery of new N-terminal and C-terminal PTH variants and the development of selected reaction monitoring (SRM)-based immunoassays specifically designed for the detection of full-length PTH [amino acid (aa)1-84] and 2 N terminal variants, aa7-84 and aa34-84. METHODS: Preparation of mass spectrometric immunoassay pipettor tips and MALDI-TOF mass spectrometric analysis were carried out as previously described. We used novel software to develop SRM assays on a triple-quadrupole mass spectrometer. Heavy isotope-labeled versions of target peptides were used as internal standards. RESULTS: Top-down analysis of samples from healthy individuals and renal failure patients revealed numerous PTH variants, including previously unidentified aa28-84, aa48-84, aa34-77, aa37-77, and aa38-77. Quantitative SRM assays were developed for PTH1-84, PTH7-84, and variant aa34-84. Peptides exhibited linear responses (R(2) = 0.90-0.99) relative to recombinant human PTH concentration limits of detection for intact PTH of 8 ng/L and limits of quantification of 16-31 ng/L depending on the peptide. Standard error of analysis for all triplicate measurements was 3%-12% for all peptides, with <5% chromatographic drift between replicates. The CVs of integrated areas under the curve for 54 separate measurements of heavy peptides were 5%-9%. CONCLUSIONS: Mass spectrometric immunoassays identified new clinical variants of PTH and provided a quantitative assay for these and previously identified forms of PTH. PMID- 20022980 TI - Automated detection of inaccurate and imprecise transitions in peptide quantification by multiple reaction monitoring mass spectrometry. AB - BACKGROUND: Multiple reaction monitoring mass spectrometry (MRM-MS) of peptides with stable isotope-labeled internal standards (SISs) is increasingly being used to develop quantitative assays for proteins in complex biological matrices. These assays can be highly precise and quantitative, but the frequent occurrence of interferences requires that MRM-MS data be manually reviewed, a time-intensive process subject to human error. We developed an algorithm that identifies inaccurate transition data based on the presence of interfering signal or inconsistent recovery among replicate samples. METHODS: The algorithm objectively evaluates MRM-MS data with 2 orthogonal approaches. First, it compares the relative product ion intensities of the analyte peptide to those of the SIS peptide and uses a t-test to determine if they are significantly different. A CV is then calculated from the ratio of the analyte peak area to the SIS peak area from the sample replicates. RESULTS: The algorithm identified problematic transitions and achieved accuracies of 94%-100%, with a sensitivity and specificity of 83%-100% for correct identification of errant transitions. The algorithm was robust when challenged with multiple types of interferences and problematic transitions. CONCLUSIONS: This algorithm for automated detection of inaccurate and imprecise transitions (AuDIT) in MRM-MS data reduces the time required for manual and subjective inspection of data, improves the overall accuracy of data analysis, and is easily implemented into the standard data analysis work flow. AuDIT currently works with results exported from MRM-MS data processing software packages and may be implemented as an analysis tool within such software. PMID- 20022982 TI - Comparison of multiplex immunoassay platforms. AB - BACKGROUND: Candidate biomarkers discovered with high-throughput proteomic techniques (along with many biomarkers reported in the literature) must be rigorously validated. The simultaneous quantitative assessment of multiple potential biomarkers across large cohorts presents a major challenge to the field. Multiplex immunoassays represent a promising solution, with the potential to provide quantitative data via parallel analyses. These assays also require substantially less sample and reagents than the traditional ELISA (which is further limited by its ability to measure only a single antigen). We have measured the reproducibility, reliability, robustness, accuracy, and throughput of commercially available multiplex immunoassays to ascertain their suitability for serum biomarker analysis and validation. METHODS: Assay platforms MULTI-ARRAY (Meso Scale Discovery), Bio-Plex (Bio-Rad Laboratories), A(2) (Beckman Coulter), FAST Quant (Whatman Schleicher & Schuell BioScience), and FlowCytomix (Bender MedSystems) were selected as representative examples of technologies currently used for high-throughput immunoanalysis. All assays were performed according to protocols specified by the manufacturers and with the reagents (diluents, calibrators, blocking reagents, and detecting-antibody mixtures) included with their kits. RESULTS: The quantifiable interval determined for each assay and antigen was based on precision (CV < 25%) and percentage recovery (measured concentration within 20% of the actual concentration). The MULTI-ARRAY and Bio Plex assays had the best performance with the lowest limits of detection, and the MULTI-ARRAY system had the most linear signal output over the widest concentration range (10(5) to 10(6)). Cytokine concentrations in unspiked and cytokine-spiked serum samples from healthy individuals were further investigated with the MULTI-ARRAY and Bio-Plex assays. CONCLUSIONS: The MULTI-ARRAY and Bio Plex multiplex immunoassay systems are the most suitable for biomarker analysis or quantification. PMID- 20022983 TI - New serology assays can detect gluten sensitivity among enteropathy patients seronegative for anti-tissue transglutaminase. AB - BACKGROUND: Some patients with celiac disease (CD) may be seronegative with the commonly used test for IgA anti-tissue transglutaminase (anti-tTG) antibodies. Our aim was to explore whether newer assays incorporating synthetic deamidated gliadin-related peptides (DGPs) or other TG isoenzymes as antigen are useful for detecting gluten sensitivity in IgA anti-tTG-seronegative patients. METHODS: We assayed serum samples obtained at diagnosis from (a) anti-tTG-seronegative patients with a CD-like enteropathy (n = 12), (b) skin biopsy-proven dermatitis herpetiformis (DH) patients (n = 26), and (c) IgA anti-tTG-positive CD patients (n = 26). All patients had typical total IgA concentrations. All patients underwent intestinal biopsy and serum testing for (a) detection of IgA and IgG isotypes of both anti-DGP and anti-tTG in a single assay (tTG/DGP Screen; INOVA Diagnostics), (b) simultaneous detection of both IgA and IgG anti-DGP antibody isotypes (DGP Dual; INOVA Diagnostics), and (c) detection of antibodies to transglutaminase 3 (TG3) or transglutaminase 6 (TG6). RESULTS: All anti-tTG seropositive patients also tested positive in anti-DGP assays. Overall, tTG/DGP Screen detected 6 (31.6%) of the 19 anti-tTG seronegatives, and anti-DGP Dual produced positive results in 5 (26.3%) of these cases. Whereas both assays detected 2 anti-tTG-negative DH patients with partial villous atrophy, they were positive in only 2 of the 5 cases with no histologically discernible mucosal damage. Testing for antibodies to TG3 and TG6 identified 7 (36.8%) of the 19 anti tTG-negative patients, 5 of which were also positive for anti-DGP. CONCLUSIONS: Detection of anti-DGP with tTG/DGP Screen or anti-DGP Dual, or detection of antibodies to other TG isoenzymes, enhances the sensitivity for detecting gluten sensitivity among non-IgA- deficient, anti-tTG-seronegative patients with CD-like enteropathy. PMID- 20022984 TI - IgG antibodies against deamidated gliadin peptides for diagnosis of celiac disease in patients with IgA deficiency. AB - BACKGROUND: Assays for IgG antibodies against deamidated gliadin (IgG-anti-dGli) are comparable in performance with tests detecting IgA antibodies against tissue transglutaminase (IgA-anti-tTG) in diagnosing celiac disease (CD). IgA-anti-tTG are absent in IgA deficiency, a condition often associated with CD. In IgA deficiency, IgG-anti-tTG, which have a lower overall diagnostic accuracy, are routinely measured. We examined whether IgG-anti-dGli would be useful for diagnosing CD in patients with IgA deficiency. METHODS: We studied 34 IgA deficient CD patients, 185 IgA-competent newly diagnosed children with CD, 316 children without CD, 400 adult blood donors, and 6 control IgA-deficient individuals without CD. Anti-dGli and anti-tTG were measured by ELISA, and endomysium antibodies (EmA) were measured by immunofluorescence on monkey esophagus (IgA as well as IgG class for all antibodies). We calculated diagnostic sensitivity (percentage of patients above cutoff with 95% CIs) according to age specific cutoffs for 95% diagnostic specificity and according to cutoffs proposed by the manufacturer of the assays. RESULTS: No IgA-deficient CD patients were positive for any IgA-based antibody assay. Diagnostic sensitivity of IgG-anti-tTG was 91.2% (95% CI 76.3%-97.7%) according to age-specific cutoffs and 82.4% (66.1% 92.0%) according to manufacturer cutoffs. The diagnostic sensitivity of IgG-EmA was 75.8% (58.8%-87.4%) and the sensitivity of IgG-anti-dGli was 88.2% (72.8% 95.9%) according to both cutoffs. CONCLUSIONS: IgG-anti-dGli and IgG-anti-tTG have comparable diagnostic sensitivities for IgA-deficient celiac patients. IgG anti-dGli may be useful for diagnosing CD in IgA-deficient patients. PMID- 20022985 TI - Integration of proteomic-based tools for improved biomarkers of myocardial injury. AB - BACKGROUND: Given the mounting evidence in favor of early pharmacologic and catheter-based interventions for patients across the spectrum of acute coronary syndromes, discovering novel diagnostically sensitive and specific biomarkers that provide biochemical proof of early or reversible myocardial injury could have a substantial positive impact on patient care. CONTENT: To address unmet needs in disease biomarkers, investigators have turned to proteomics approaches. We describe advances in proteomics discovery technologies based on liquid chromatography-tandem mass spectrometry that facilitate the unbiased analysis of low-abundance blood proteins. We detail the development of emerging techniques to enhance the biomarker verification process, such as accurate inclusion mass screening, stable isotope dilution-multiple reaction monitoring-mass spectrometry (SID-MRM-MS), and stable isotope standards with capture by antipeptide antibodies, which combines the advantages of specific immunoaffinity enrichment of a target peptide with the structural specificity and quantitative capabilities of SID-MRM-MS. We highlight new assays incorporating these techniques for troponin I, a representative low-abundance cardiac biomarker, and interleukin-33, an emerging novel marker of myocardial stress for which no existing ELISA exists. We demonstrate that troponin I and interleukin-33 peptides have a linear, dynamic range spanning 4 orders of magnitude and limits of detection of approximately 0.5 microg/L back-calculated to the protein concentration. CONCLUSIONS: There remain important unmet diagnostic and prognostic needs in cardiology. Advances in technology may allow proteomics to play a vital role in the discovery and validation of novel biomarkers to help fill those needs. PMID- 20022986 TI - Medical treatment predicts mortality after hip fracture. AB - BACKGROUND: The mortality after hip fracture has remained high and stable the past 50 years despite improved surgical treatment. The aim of this study was to identify medications and medical factors associated with mortality after hip fracture. METHODS: This is a prospective observational study with median observation time of 21 months. Three hundred and sixty-four patients, mean age 83.4 years and 75.8% women, were enrolled. Information on comorbidity, medications, surgery, and clinical findings were collected at the time of fracture. Information on cause and time of death was obtained from the Norwegian Cause of Death Register. RESULTS: Six risk factors and one protective factor were identified by Cox proportional hazards model adjusted for propensity score: the use of diuretics (adjusted hazard ratio [HR] = 4.03, 95% confidence interval [CI] = 2.13-7.64), history of coronary heart disease (CHD) (HR = 2.61, CI = 1.37 4.98), male sex (HR = 2.32, CI = 1.27-4.24), Barthel Index < or = 18/20 (HR = 2.48, CI = 1.23-5.01), heart rate > 100 on admission (HR = 2.47, CI = 1.18-5.14), body mass index < or = 20 (HR = 1.94, CI = 1.13-3.34), and the use of statins (HR = 0.23, CI = 0.08-0.68). Patients using diuretics had increased risk of death from all causes, including death from CHD, chronic obstructive pulmonary disease, and falls or other accidents. CONCLUSIONS: The use of diuretics is the strongest predictor of mortality, followed by CHD at the time of fracture, whereas the use of statins is associated with improved survival. Future research is needed to evaluate whether improved diagnosis and management of CHD and congestive heart failure among hip fracture patients would improve survival. PMID- 20022988 TI - Ethnic differences in incidence of type 1 diabetes among second-generation immigrants and adoptees from abroad. AB - OBJECTIVE: The incidence of type 1 diabetes shows a large variation worldwide, but whether the causes are environmental or genetic has not been settled. We examine here the incidence of type 1 diabetes among second-generation immigrants and adoptees from abroad to disentangle genetic/ethnic vs. environmental influence, assuming adoptees from abroad have similar environmental exposures compared with the native Swedes, with the only difference in their genetic background. METHODS: Second-generation immigrants and adoptees from abroad were retrieved from the MIGMED2 database, and they were followed up until the diagnosis of type 1 diabetes, death, or the end of study. Standardized incidence ratios (SIRs) were calculated for type 1 diabetes among these immigrants compared with native Swedes. RESULTS: A total of 1,050,569 children were defined as second generation immigrants and the overall SIR of type 1 diabetes was significantly decreased. A decreased risk was observed for all countries of origin, with an exception for children with parents from Finland. A total of 51,557 children born in foreign countries were adopted by Swedes. Adoptees from Eastern Europe, Soviet countries, India, Pakistan, Afghanistan, East and Southeast Asia, Chile, and other Central and South American countries had a significantly decreased SIR. CONCLUSIONS: The decreased incidence of type 1 diabetes observed in some second generation immigrants and adoptees from abroad strongly suggests that ethnic genetic heterogeneity could play an important role on type 1 diabetes. PMID- 20022987 TI - The effect of branched chain amino acids on skeletal muscle mitochondrial function in young and elderly adults. AB - CONTEXT: A reduction in maximal mitochondrial ATP production rate (MAPR) and mitochondrial DNA (mtDNA) abundance occurs with age in association with muscle weakness and reduced endurance in elderly people. Branched chain amino acids (BCAA) have been extensively used to improve physical performance. OBJECTIVE: The objective was to determine whether an 8-h infusion of BCAA enhances MAPR equally in healthy young and elderly adults. METHODS: Using a crossover study design, we compared the effect BCAA vs. saline infusion in 12 young (23.0 +/- 0.8 yr) and 12 elderly (70.7 +/- 1.1 yr) participants matched for sex and body mass index. Skeletal muscle MAPR and mtDNA abundance were measured in muscle biopsy samples obtained before and at the end of the 8-h infusion. RESULTS: In young participants, MAPR with the substrates glutamate plus malate (supplying electrons to complex I) and succinate plus rotenone (complex II) increased in response to BCAA infusion, relative to a decline in MAPR in response to the saline infusion. In contrast, MAPR was unaffected by BCAA infusion in the elderly participants. Moreover, mtDNA abundance was lower in the elderly compared with the young participants but was unaffected by the BCAA infusion. Insulin and C-peptide concentrations declined over time during the saline infusion, but these declines were prevented by the BCAA infusion. CONCLUSIONS: BCAA increased skeletal muscle MAPR in the young participants in comparison with saline, but this effect was not seen in the elderly participants indicating, that unlike in the young, BCAA does not increase muscle mitochondrial function in the elderly. PMID- 20022989 TI - The sulfaphenazole-sensitive pathway acts as a compensatory mechanism for impaired nitric oxide availability in patients with primary hyperparathyroidism. Effect of surgical treatment. AB - OBJECTIVE: The aim of this study was to assess whether patients with primary hyperparathyroidism (PHPT) show reduced endothelial function and to determine the mechanisms involved. The impact of parathyroidectomy (PTx) on endothelial function was also assessed. BACKGROUND: Endothelial dysfunction is reported in patients with PHPT, but the mechanisms involved are unknown. METHODS: We evaluated forearm blood flow changes (strain gauge plethysmography) induced by intraarterial acetylcholine or sodium nitroprusside in 17 PHPT women and 17 age matched controls. Nitric oxide (NO) availability and oxidative stress were studied by repeating acetylcholine during intraarterial infusion of L-N(G) monomethyl arginine (L-NMMA, a NO synthase inhibitor) and ascorbic acid (an oxidative stress scavenger). The role of cytochrome P450 epoxygenase (CYP 2C9) derived endothelium-derived hyperpolarizing factor (EDHF) was assessed by repeating acetylcholine under intraarterial sulfaphenazole. In six PHPT patients, the study was repeated 12 months after successful PTx. RESULTS: Responses to sodium nitroprusside and acetylcholine were similar in PHPT patients and controls. L-NMMA inhibited the response to acetylcholine in controls (P < 0.001), whereas it had no effect in PHPT patients. In both groups, ascorbic acid failed to affect acetylcholine. Sulfaphenazole administration, although not affecting vasodilation to acetylcholine in controls, blunted the response to acetylcholine in PHPT patients (P < 0.005). After PTx, the inhibitory effect of L-NMMA on acetylcholine was restored (P < 0.001), and the inhibitory effect of sulfaphenazole on acetylcholine was abrogated. CONCLUSIONS: PHPT patients show compromised NO availability, whereas oxidative stress generation is not involved. A compensatory CYP 2C9-derived EDHF pathway is activated to sustain endothelium dependent vasodilation. This PHPT-related endothelial dysfunction is reversed after PTx. PMID- 20022990 TI - Prevention of bone loss by zoledronic acid in premenopausal women undergoing adjuvant chemotherapy persist up to one year following discontinuing treatment. AB - CONTEXT: Adjuvant chemotherapy is associated with significant reductions in bone mineral density (BMD) in premenopausal women with breast cancer (BC) that is prevented with zoledronic acid (ZA) every 3 months for 1 yr. OBJECTIVE: The aim of the study was to examine the effect on BMD of discontinuing ZA during the subsequent year. DESIGN: We conducted a randomized, double-blind trial. PATIENTS: Premenopausal women (mean age, 42 yr) undergoing adjuvant chemotherapy for BC participated in the study. INTERVENTION: ZA (4 mg iv every 3 months) vs. placebo was administered for 12 months. OUTCOME MEASURES: We measured percentage change in BMD and bone turnover markers at 12 and 24 months (1 yr after last infusion). RESULTS: Of 101 women randomized, 85 completed 12-month and 62 completed 24-month evaluations. In the placebo group, serum C-telopeptide (CTX) increased progressively over the first 12 months, returned toward baseline but remained significantly above baseline by 24 months. Lumbar spine BMD decreased from baseline by 5.5% at 12 and 6.3% at 24 months. Similarly, by 24 months, total hip and femoral neck BMD declined by 2.6 and 2.4%, respectively. In ZA patients, BMD remained stable (P < 0.0001 compared to placebo). Serum CTX declined significantly by 6 months, but returned to baseline by 12 months, remaining there at 24 months. CONCLUSIONS: Premenopausal women receiving chemotherapy for BC sustained significant bone loss during the first year, without recovery during the second year. ZA effectively prevented bone loss during the first year of chemotherapy. BMD remained stable 1 yr after completion of ZA. Serum CTX increased significantly by 12 and 24 months. More frequent administration may be required to suppress bone resorption in this patient population. PMID- 20022991 TI - A comparative phenotypic study of kallmann syndrome patients carrying monoallelic and biallelic mutations in the prokineticin 2 or prokineticin receptor 2 genes. AB - CONTEXT: Both biallelic and monoallelic mutations in PROK2 or PROKR2 have been found in Kallmann syndrome (KS). OBJECTIVE: The objective of the study was to compare the phenotypes of KS patients harboring monoallelic and biallelic mutations in these genes. DESIGN AND PATIENTS: We studied clinical and endocrine features that reflect the functioning of the pituitary-gonadal axis, and the nonreproductive phenotype, in 55 adult KS patients (42 men and 13 women), of whom 41 had monoallelic mutations and 14 biallelic mutations in PROK2 or PROKR2. RESULTS: Biallelic mutations were associated with more frequent cryptorchidism (70% vs. 34%, P < 0.05) and microphallus (90% vs. 28%, P < 0.001) and lower mean testicular volume (1.2 +/- 0.4 vs. 4.5 +/- 6.0 ml; P < 0.01) in male patients. Likewise, the testosterone level as well as the basal FSH level and peak LH level under GnRH-stimulation were lower in males with biallelic mutations (0.2 +/- 0.1 vs. 0.7 +/- 0.8 ng/ml; P = 0.05, 0.3 +/- 0.1 vs. 1.8 +/- 3.0 IU/liter; P < 0.05, and 0.8 +/- 0.8 vs. 5.2 +/- 5.5 IU/liter; P < 0.05, respectively). Nonreproductive, nonolfactory anomalies were rare in both sexes and were never found in patients with biallelic mutations. The mean body mass index of the patients (23.9 +/- 4.2 kg/m(2) in males and 26.3 +/- 6.6 kg/m(2) in females) did not differ significantly from that of gender-, age-, and treatment-matched KS individuals who did not carry a mutation in PROK2 or PROKR2. Finally, circadian cortisol levels evaluated in five patients, including one with biallelic PROKR2 mutations, were normal in all cases. CONCLUSION: Male patients carrying biallelic mutations in PROK2 or PROKR2 have a less variable and on average a more severe reproductive phenotype than patients carrying monoallelic mutations in these genes. Nonreproductive, nonolfactory clinical anomalies associated with KS seem to be restricted to patients with monoallelic mutations. PMID- 20022992 TI - Metabolic, cardiovascular, and cerebrovascular outcomes in growth hormone deficient subjects with previous cushing's disease or non-functioning pituitary adenoma. AB - CONTEXT: Previous exposure to hypercortisolism due to Cushing's disease (CD) may adversely affect long-term metabolic and cardiovascular outcomes. In particular, metabolic and cardiovascular outcomes of patients with previous CD who require GH replacement have not been fully established. OBJECTIVE: The aim of the study was to compare the prevalence and incidence of metabolic syndrome (Adult Treatment Panel III criteria), diabetes mellitus, cardiovascular disease, and cerebrovascular disease in GH-treated subjects with previous CD with GH-treated subjects with previous nonfunctioning pituitary adenoma (NFPA). DESIGN: We conducted post hoc analysis of the observational Hypopituitary Control and Complications Study conducted at 362 international centers (1995-2006). SUBJECTS: We studied adult-onset GH-deficient subjects with previous CD (n = 160) or NFPA (n = 879). All subjects received GH replacement therapy and were GH naive at enrollment. Multiple pituitary deficits were prevalent in both groups. MAIN OUTCOME MEASURES: We measured the prevalence and incidence of metabolic syndrome, diabetes mellitus, cardiovascular disease, and cerebrovascular disease at baseline and at 3 yr, standardized for age and sex differences between groups. RESULTS: Compared with subjects with previous NFPA, subjects with previous CD had a significantly greater 3-yr incidence of metabolic syndrome (CD, 23.4%; NFPA, 9.2%; P = 0.01), baseline (CD, 6.3%; NFPA, 2.2%; P < 0.01) and 3-yr (CD, 7.6%; NFPA, 3.9%; P = 0.04) prevalence of cardiovascular disease, and baseline (CD, 6.4%; NFPA, 1.8%; P = 0.03) and 3-yr (CD, 10.2%; NFPA, 2.9%; P = 0.01) prevalence of cerebrovascular disease. CONCLUSIONS: Previous hypercortisolism may predispose GH-treated, GH-deficient subjects with prior CD to an increased risk of metabolic syndrome, cardiovascular disease, and cerebrovascular disease. PMID- 20022993 TI - Mental practice for relearning locomotor skills. AB - Over the past 2 decades, much work has been carried out on the use of mental practice through motor imagery for optimizing the retraining of motor function in people with physical disabilities. Although much of the clinical work with mental practice has focused on the retraining of upper-extremity tasks, this article reviews the evidence supporting the potential of motor imagery for retraining gait and tasks involving coordinated lower-limb and body movements. First, motor imagery and mental practice are defined, and evidence from physiological and behavioral studies in healthy individuals supporting the capacity to imagine walking activities through motor imagery is examined. Then the effects of stroke, spinal cord injury, lower-limb amputation, and immobilization on motor imagery ability are discussed. Evidence of brain reorganization in healthy individuals following motor imagery training of dancing and of a foot movement sequence is reviewed, and the effects of mental practice on gait and other tasks involving coordinated lower-limb and body movements in people with stroke and in people with Parkinson disease are examined. Lastly, questions pertaining to clinical assessment of motor imagery ability and training strategies are discussed. PMID- 20022994 TI - Similarity of joint kinematics and muscle demands between elliptical training and walking: implications for practice. AB - BACKGROUND: People with physical disabilities often face barriers to regaining walking ability and fitness after discharge from rehabilitation. Physical therapists are uniquely positioned to teach clients the knowledge and skills needed to exercise on functionally relevant equipment available in the community, such as elliptical trainers. However, therapeutic use is hindered by a lack of empirical information. OBJECTIVE: The purpose of this study was to examine joint kinematics and muscle activation recorded during walking and elliptical training to provide evidence-based data to guide clinical decision making. DESIGN: This was a prospective, controlled laboratory study using a repeated-measures design. METHODS: Twenty adults free from impairments that might hinder gait participated. After familiarization procedures, subjects walked and trained on 4 elliptical devices while kinematic, electromyographic (EMG), and stride characteristic data were recorded. RESULTS: Movement similarities between elliptical training and walking were supported by the documentation of relatively high coefficients of multiple correlation for the hip (.85-.89), thigh (.92-.94), knee (.87-.89) and, to a lesser extent, the ankle (.57-.71). Significantly greater flexion was documented at the trunk, pelvis, hip, and knee during elliptical training than during walking. One of the elliptical trainers most closely simulated sagittal plane walking kinematics, as determined from an assessment of key variables. During elliptical training, gluteus maximus and vastus lateralis muscle activation were increased; medial hamstring, gastrocnemius, soleus, and tibialis anterior muscle activation were decreased; and gluteus medius and lateral hamstring muscle activation were relatively unchanged compared with muscle activation of those muscles in walking. On the basis of EMG findings, no elliptical trainer clearly emerged as the best for simulating gait. LIMITATIONS: To date, only 4 elliptical trainers have been studied, and the contributions of the upper extremities to movement have not been quantified. CONCLUSIONS: Although one of the elliptical trainers best simulated sagittal-plane walking kinematics, EMG analysis failed to identify one clearly superior device. This research provides evidence-based data to help guide clinical decision making related to the use of elliptical trainers across the health care continuum and into the community. PMID- 20022996 TI - Gait parameters associated with responsiveness to treadmill training with body weight support after stroke: an exploratory study. AB - BACKGROUND: Task-specific training programs after stroke improve walking function, but it is not clear which biomechanical parameters of gait are most associated with improved walking speed. OBJECTIVE: The purpose of this study was to identify gait parameters associated with improved walking speed after a locomotor training program that included body-weight-supported treadmill training (BWSTT). DESIGN: A prospective, between-subjects design was used. METHODS: Fifteen people, ranging from approximately 9 months to 5 years after stroke, completed 1 of 3 different 6-week training regimens. These regimens consisted of 12 sessions of BWSTT alternated with 12 sessions of: lower-extremity resistive cycling; lower-extremity progressive, resistive strengthening; or a sham condition of arm ergometry. Gait analysis was conducted before and after the 6 week intervention program. Kinematics, kinetics, and electromyographic (EMG) activity were recorded from the hemiparetic lower extremity while participants walked at a self-selected pace. Changes in gait parameters were compared in participants who showed an increase in self-selected walking speed of greater than 0.08 m/s (high-response group) and in those with less improvement (low response group). RESULTS: Compared with participants in the low-response group, those in the high-response group displayed greater increases in terminal stance hip extension angle and hip flexion power (product of net joint moment and angular velocity) after the intervention. The intensity of soleus muscle EMG activity during walking also was significantly higher in participants in the high response group after the intervention. LIMITATIONS: Only sagittal-plane parameters were assessed, and the sample size was small. CONCLUSIONS: Task specific locomotor training alternated with strength training resulted in kinematic, kinetic, and muscle activation adaptations that were strongly associated with improved walking speed. Changes in both hip and ankle biomechanics during late stance were associated with greater increases in gait speed. PMID- 20022995 TI - Meaningful gait speed improvement during the first 60 days poststroke: minimal clinically important difference. AB - BACKGROUND: When people with stroke recover gait speed, they report improved function and reduced disability. However, the minimal amount of change in gait speed that is clinically meaningful and associated with an important difference in function for people poststroke has not been determined. OBJECTIVE: The purpose of this study was to determine the minimal clinically important difference (MCID) for comfortable gait speed (CGS) associated with an improvement in the modified Rankin Scale (mRS) score for people between 20 to 60 days poststroke. DESIGN: This was a prospective, longitudinal, cohort study. METHODS: The participants in this study were 283 people with first-time stroke prospectively enrolled in the ongoing Locomotor Experience Applied Post Stroke (LEAPS) multi-site randomized clinical trial. Comfortable gait speed was measured and mRS scores were obtained at 20 and 60 days poststroke. Improvement of >or=1 on the mRS was used to detect meaningful change in disability level. RESULTS: Mean (SD) CGS was 0.18 (0.16) m/s at 20 days and 0.39 (0.22) m/s at 60 days poststroke. Among all participants, 47.3% experienced an improvement in disability level >or=1. The MCID was estimated as an improvement in CGS of 0.16 m/s anchored to the mRS. LIMITATIONS: Because the mRS is not a gait-specific measure of disability, the estimated MCID for CGS was only 73.9% sensitive and 57.0% specific for detecting improvement in mRS scores. CONCLUSIONS: We estimate that the MCID for gait speed among patients with subacute stroke and severe gait speed impairments is 0.16 m/s. Patients with subacute stroke who increase gait speed >or=0.16 m/s are more likely to experience a meaningful improvement in disability level than those who do not. Clinicians can use this reference value to develop goals and interpret progress in patients with subacute stroke. PMID- 20022997 TI - Daily stepping in individuals with motor incomplete spinal cord injury. AB - BACKGROUND: In individuals with motor incomplete spinal cord injury (SCI), ambulatory function determined in the clinical setting is related to specific measures of body structure and function and activity limitations, although few studies have quantified the relationship of these variables with daily stepping (steps/day). OBJECTIVE: The aim of this study was to quantify daily stepping in ambulatory individuals with SCI and its relationship with clinical walking performance measures and specific demographics, impairments, and activity limitations. DESIGN: A cross-sectional study was performed to estimate relationships among clinical variables to daily stepping in self-identified community versus non-community (household) walkers. METHODS: Average daily stepping was determined in 50 people with chronic, motor incomplete SCI. Data for clinical and self-report measures of walking performance also were collected, and their associations with daily stepping were analyzed using correlation and receiver operating characteristic (ROC) analyses. Relationships between daily stepping and the measures of demographics, impairments, and activity limitations were identified using correlation and regression analyses. RESULTS: The ROC analyses revealed a significant discriminative ability between self-reported community and non-community walkers using clinical gait measures and daily stepping. Stepping activity generally was low throughout the sample tested, however, with an average of approximately 2,600 steps/day. Knee extension strength (force-generating capacity) and static balance were the primary variables related to daily stepping, with metabolic efficiency and capacity and balance confidence contributing to a lesser extent. LIMITATIONS: The small sample size and use of specific impairment-related measures were potential limitations of the study. CONCLUSIONS: Daily stepping is extremely limited in individuals with incomplete SCI, with a potentially substantial contribution of impairments in knee extension strength and balance. PMID- 20022998 TI - Striding out with Parkinson disease: evidence-based physical therapy for gait disorders. AB - Although Parkinson disease (PD) is common throughout the world, the evidence for physical therapy interventions that enable long-term improvement in walking is still emerging. This article critiques the major physical therapy approaches related to gait rehabilitation in people with PD: compensatory strategies, motor skill learning, management of secondary sequelae, and education to optimize physical activity and reduce falls. The emphasis of this review is on gait specifically, although balance and falls are of direct importance to gait and are addressed in that context. Although the researchers who have provided the evidence for these approaches grounded their studies on different theoretical paradigms, each approach is argued to have a valid place in the comprehensive management of PD generally and of gait in particular. The optimal mix of interventions for each individual varies according to the stage of disease progression and the patient's preferred form of exercise, capacity for learning, and age. PMID- 20022999 TI - Can strength training predictably improve gait kinematics? A pilot study on the effects of hip and knee extensor strengthening on lower-extremity alignment in cerebral palsy. AB - BACKGROUND: Computer simulations have demonstrated that excessive hip and knee flexion during gait, as frequently seen in ambulatory children with cerebral palsy (CP), can reduce the ability of muscles to provide antigravity support and increase the tendency of hip muscles to internally rotate the thigh. These findings suggest that therapies for improving upright posture during gait also may reduce excessive internal rotation. OBJECTIVE: The goal of this study was to determine whether strength training can diminish the degree of crouched, internally rotated gait in children with spastic diplegic CP. DESIGN: This was a pilot prospective clinical trial. METHODS: Eight children with CP participated in an 8-week progressive resistance exercise program, with 3-dimensional gait analysis and isokinetic testing performed before and after the program. Secondary measures included passive range of motion, the Ashworth Scale, and the PedsQL CP Module. To identify factors that may have influenced outcome, individual and subgroup data were examined for patterns of change within and across variables. RESULTS: Strength (force-generating capacity) increased significantly in the left hip extensors, with smaller, nonsignificant mean increases in the other 3 extensor muscle groups, yet kinematic and functional outcomes were inconsistent. The first reported subject-specific computer simulations of crouch gait were created for one child who showed substantial benefit to examine the factors that may have contributed to this outcome. LIMITATIONS: The sample was small, with wide variability in outcomes. CONCLUSIONS: Strength training may improve walking function and alignment in some patients for whom weakness is a major contributor to their gait deficits. However, in other patients, it may produce no change or even undesired outcomes. Given the variability of outcomes in this and other strengthening studies in CP, analytical approaches to determine the sources of variability are needed to better identify those individuals who are most likely to benefit from strengthening. PMID- 20023000 TI - How does explicit prioritization alter walking during dual-task performance? Effects of age and sex on gait speed and variability. AB - BACKGROUND: Previous studies have demonstrated that the performance of a secondary task during walking alters gait. OBJECTIVE: This study investigated the effects of task prioritization on walking in young and older adults to evaluate the "default" prioritization scheme used, the flexibility to alter prioritization and cortical resources allocated to gait and a secondary cognitive task, and any age-associated changes in these abilities. DESIGN: A cross-sectional study that explicitly altered the focus of attention was used to investigate the effects of prioritization in young and older adults who were healthy. METHODS: Gait speed and gait variability were evaluated in young adults (n=40) and older adults (n=17) who were healthy, both during usual walking and under 3 dual-task conditions: (1) no specific prioritization instructions, (2) prioritization of gait, and (3) prioritization of the cognitive task. RESULTS: Young adults significantly increased gait speed in the gait prioritization condition compared with gait speed in the no-instruction condition; a similar tendency was seen in the older adults. Gait speed was reduced when priority was given to the cognitive task in both age groups; however, this effect was less dramatic in the older adults. In the young adults, prioritization of gait tended to have different effects on gait speed among both men and women. In the older adults, but not in the young adults, all dual-task conditions produced increased gait variability, whereas prioritization did not alter this gait feature. LIMITATIONS: The sample size and the relative homogeneity of the older adults could be considered as possible limitations of the study. CONCLUSIONS: Even among young adults, the effects of secondary, cognitive tasks on gait speed are strongly influenced by prioritization. This finding was less significant in the older adults, suggesting that there is an age-associated decline in the ability to flexibly allocate attention to gait. Somewhat surprisingly, when prioritization was not explicitly instructed, gait speed in both young and older adults most closely resembled that of the condition when they were instructed to focus attention on the cognitive task. PMID- 20023001 TI - Neurophysiologic and rehabilitation insights from the split-belt and other locomotor adaptation paradigms. AB - Locomotion is incredibly flexible. Humans are able to stay upright and navigate long distances in the face of ever-changing environments and varied task demands, such as walking while carrying a heavy object or in thick mud. The focus of this review is a behavior that is critical for this flexibility: motor adaptation. Adaptation is defined here as the process of adjusting a movement to new demands through trial-and-error practice. A key feature of adaptation is that more practice without the new demand is required to return the movement to its original state. Thus, motor adaptation is a short-term motor learning process. Several studies have been undertaken to determine how humans adapt walking to novel circumstances. Many of these studies have examined locomotor adaptation using a split-belt treadmill. The results of these studies of people who were healthy and people with neurologic damage suggest that the cerebellum is required for normal adaptation of walking and that the role of cerebral structures may be less critical. They also suggest that intersegmental and interlimb coordination is critical but readily adaptable to accommodate changes in the environment. Locomotor adaptation also can be used to determine the walking potential of people with specific neurologic deficits. For instance, split-belt and limb weighting locomotor adaptation studies show that adults with chronic stroke are capable of improving weight-bearing and spatiotemporal symmetry, at least temporarily. Our challenge as rehabilitation specialists is to intervene in ways that maximize this capacity. PMID- 20023002 TI - Dynamic principles of gait and their clinical implications. AB - A healthy gait pattern depends on an array of biomechanical features, orchestrated by the central nervous system for economy and stability. Injuries and other pathologies can alter these features and result in substantial gait deficits, often with detrimental consequences for energy expenditure and balance. An understanding of the role of biomechanics in the generation of healthy gait, therefore, can provide insight into these deficits. This article examines the basic principles of gait from the standpoint of dynamic walking, an approach that combines an inverted pendulum model of the stance leg with a pendulum model of the swing leg and its impact with the ground. The heel-strike at the end of each step has dynamic effects that can contribute to a periodic gait and its passive stability. Biomechanics, therefore, can account for much of the gait pattern, with additional motor inputs that are important for improving economy and stability. The dynamic walking approach can predict the consequences of disruptions to normal biomechanics, and the associated observations can help explain some aspects of impaired gait. This article reviews the basic principles of dynamic walking and the associated experimental evidence for healthy gait and then considers how the principles may be applied to clinical gait pathologies. PMID- 20023003 TI - Cognitive load and dual-task performance during locomotion poststroke: a feasibility study using a functional virtual environment. AB - BACKGROUND: Gait and cognitive functions can deteriorate during dual tasking, especially in people with neurological deficits. Most studies examining the simultaneous effects of dual tasking on motor and cognitive aspects were not performed in ecological environments. Using virtual reality technology, functional environments can be simulated to study dual tasking. OBJECTIVES: The aims of this study were to test the feasibility of using a virtual functional environment for the examination of dual tasking and to determine the effects of dual tasking on gait parameters in people with stroke and age-matched controls who were healthy. DESIGN: This was a cross-sectional observational study. METHODS: Twelve community-dwelling older adults with stroke and 10 age-matched older adults who were healthy participated in the study. Participants walked on a self-paced treadmill while viewing a virtual grocery aisle projected onto a screen placed in front of them. They were asked to walk through the aisle (single task) or to walk and select ("shop for") items according to instructions delivered before or during walking (dual tasking). RESULTS: Overall, the stroke group walked slower than the control group in both conditions, whereas both groups walked faster overground than on the treadmill. The stroke group also showed larger variability in gait speed and shorter stride length than the control group. There was a general tendency to increase gait speed and stride length during dual-task conditions; however, a significant effect of dual tasking was found only in one dual-task condition for gait speed and stride duration variability. All participants were able to complete the task with minimal mistakes. LIMITATIONS: The small size and heterogeneity of the sample were limitations of the study. CONCLUSIONS: It is feasible to use a functional virtual environment for investigation of dual tasking. Different gait strategies, including an increase or decrease in gait speed, can be used to cope with the increase in cognitive demands required for dual tasking. PMID- 20023004 TI - Rectus femoris to gracilis muscle transfer with fractional lengthening of the vastus muscles: a treatment for adults with stiff knee gait. AB - BACKGROUND: Stiff knee gait, which may be seen in patients with upper motor neuron injury, describes a gait pattern with a relative loss of sagittal knee motion. It interferes with foot clearance during swing, often leading to inefficient compensatory mechanisms and ambulatory dysfunction. Distal rectus femoris muscle transfers and fractional lengthening of the vastus muscles have been performed in adult patients. OBJECTIVE: The purpose of this study was to describe a unique surgical technique and report on initial outcomes. DESIGN: A retrospective case-series study design was used. METHODS: The patients were adults with stiff knee gait due to stroke or traumatic brain injury who underwent distal rectus femoris muscle transfer with fractional lengthening of the vastus muscles. The patients (19 men and 18 women) had an average age of 51 years at the time of surgery. Lower-extremity examinations, clinical gait analyses, and satisfaction levels were recorded preoperatively and postoperatively. RESULTS: At a mean follow-up time of 10 months, 36 (97%) of the 37 patients were satisfied with their clinical and functional results, and the average Viosca score improved from 3.1 to 3.5. LIMITATIONS: Limitations of the study include use of a retrospective design, lack of a control group, and limited quantitative measures of gait. CONCLUSION: Distal rectus femoris muscle transfer and fractional lengthening of the vastus muscles were found to be a possible treatment for adults with stiff-knee gait caused by stroke or traumatic brain injury. PMID- 20023005 TI - Activation of nerve growth factor-induced B alpha by methylene-substituted diindolylmethanes in bladder cancer cells induces apoptosis and inhibits tumor growth. AB - Nerve growth factor-induced B (NGFI-B) genes are orphan nuclear receptors, and NGFI-B alpha (Nur77, TR3) is overexpressed in bladder tumors and bladder cancer cells compared with nontumorous bladder tissue. 1,1-Bis(3'-indolyl)-1-(p methoxyphenyl)-methane (DIM-C-pPhOCH(3)) and 1,1-bis(3'-indolyl)-1-(p phenyl)methane have previously been identified as activators of Nur77, and both compounds inhibited growth and induced apoptosis of UC-5 and KU7 bladder cancer cells. The proapoptotic effects of methylene-substituted diindolylmethanes (C DIMs) were unaffected by cotreatment with leptomycin B and were dependent on nuclear Nur77, and RNA interference with a small inhibitory RNA for Nur77 (iNur77) demonstrated that C-DIM-induced activation of apoptosis was Nur77 dependent. Microarray analysis of DIM-C-pPhOCH(3)-induced genes in UC-5 bladder cancer cells showed that this compound induced multiple Nur77-dependent proapoptotic or growth inhibitory genes including tumor necrosis factor-related apoptosis-inducing ligand (TRAIL), cystathionase, p21, p8, and sestrin-2. DIM-C pPhOCH(3) (25 mg/kg/d) also induced apoptosis and inhibited tumor growth in athymic nude mice bearing KU7 cells as xenografts, demonstrating that Nur77 active C-DIMs exhibit potential for bladder cancer chemotherapy by targeting Nur77, which is overexpressed in this tumor type. PMID- 20023006 TI - Iron chelator-mediated alterations in gene expression: identification of novel iron-regulated molecules that are molecular targets of hypoxia-inducible factor-1 alpha and p53. AB - Iron deficiency affects 500 million people, yet the molecular role of iron in gene expression remains poorly characterized. In addition, the alterations in global gene expression after iron chelation remain unclear and are important to assess for understanding the molecular pathology of iron deficiency and the biological effects of chelators. Considering this, we assessed the effect on whole genome gene expression of two iron chelators (desferrioxamine and 2-hydroxy 1-napthylaldehyde isonicotinoyl hydrazone) that have markedly different permeability properties. Sixteen genes were significantly regulated by both ligands, whereas a further 50 genes were significantly regulated by either compound. Apart from iron-mediated regulation of expression via hypoxia inducible factor-1 alpha, it was noteworthy that the transcription factor p53 was also involved in iron-regulated gene expression. Examining 16 genes regulated by both chelators in normal and neoplastic cells, five genes (APP, GDF15, CITED2, EGR1, and PNRC1) were significantly differentially expressed between the cell types. In view of their functions in tumor suppression, proliferation, and apoptosis, these findings are important for understanding the selective antiproliferative effects of chelators against neoplastic cells. Most of the genes identified have not been described previously to be iron-regulated and are important for understanding the molecular and cellular effects of iron depletion. PMID- 20023008 TI - Differential responses of epithelial Sertoli cells of the rat testis to Toll-like receptor 2 and 4 ligands: implications for studies of testicular inflammation using bacterial lipopolysaccharides. AB - The relative contribution of epithelial Sertoli cells in response to bacterial infection of the testis remains poorly characterised, since studies on inflammatory properties of these cells have invariably used unpurified lipopolysaccharide (LPS) preparations contaminated with bacterial lipopeptides. Consequently, isolated rat Sertoli cells were stimulated with either unextracted or phenol re-extracted LPS, and analysed for Toll-like receptor (TLR) 4, TLR2 and inflammatory cytokine gene expression by quantitative reverse transcription polymerase chain reaction (RT-PCR). Expression of TLR4 and its co-receptor protein myeloid differentiation (MD) 2 in Sertoli cells and testicular macrophages were similar, but Sertoli cells displayed low basal or LPS-induced expression of the TLR4 accessory protein, CD14. In Sertoli cells, unextracted LPS produced cytokine responses which were considerably greater in magnitude and duration compared with their response to purified LPS. Sertoli cells also responded to the synthetic lipopeptide, Pam(3)Cys (a TLR2 ligand) with a similar pattern of prolonged gene expression. Sertoli cells were more than 10-fold less sensitive to purified LPS than macrophages, but expressed similar levels of interleukin (IL)-1alpha and IL-6, and much greater levels of the immunoregulatory cytokine activin A, when maximally stimulated. These data demonstrate that Sertoli cells display differential cytokine responses to bacterial stimuli, mediated by both TLR2 and TLR4, that are distinct from those of testicular macrophages. PMID- 20023007 TI - ONO 3403, a synthetic serine protease inhibitor, inhibits lipopolysaccharide induced tumor necrosis factor-{alpha} and nitric oxide production and protects mice from lethal endotoxic shock. AB - ONO 3403, a new synthetic serine protease inhibitor, is a derivative of camostat mesilate and has a higher protease-inhibitory activity. The effect of ONO 3403 on lipopolysaccharide (LPS)-induced tumor necrosis factor (TNF)-alpha and nitric oxide (NO) production in RAW 264.7 macrophage-like cells was examined. ONO 3403 significantly inhibited LPS-induced TNF-alpha production at a lower concentration than camostat mesilate. It also inhibited LPS-induced NO production. Their inhibition was responsible for the reduced mRNA expression of TNF-alpha and inducible NO synthase. In LPS-stimulated cells, ONO 3403 prevented the augmentation of MyD88 expression and inhibited the phosphorylation of IkappaB alpha, stress-activated protein kinase (SAPK) and IRF-3, and the production of interferon-beta. ONO 3403 abolished the elevation of the extracellular serine protease activity in response to LPS. Further, it reduced the circulating TNF alpha level, hepatic injury and mortality in mice receiving an injection of D galactosamine and LPS. ONO 3403 was suggested to inhibit LPS-induced inflammatory responses via inactivation of MyD88-dependent and independent pathways. PMID- 20023009 TI - Functional analysis of the stability determinant AlfB of pBET131, a miniplasmid derivative of bacillus subtilis (natto) plasmid pLS32. AB - Bacillus subtilis plasmid pBET131 is a derivative of pLS32, which was isolated from a natto strain of Bacillus subtilis. The DNA region in pBET131 that confers segregational stability contains an operon consisting of three genes, of which alfA, encoding an actin-like ATPase, and alfB are essential for plasmid stability. In this work, the alfB gene product and its target DNA region were studied in detail. Transcription of the alf operon initiated from a sigma(A)-type promoter was repressed by the alfB gene product. Overproduction of AlfA was inhibitory to cell growth, suggesting that the repression of the alf operon by AlfB is important for maintaining appropriate levels of AlfA. An electrophoretic mobility shift assay and footprinting analysis with purified His-tagged AlfB showed that it bound to a DNA region containing three tandem repeats of 8-bp AT rich sequence (here designated parN), which partially overlaps the -35 sequence of the promoter. A sequence alteration in the first or third repeat did not affect the AlfB binding and plasmid stability, whereas that in the second repeat resulted in inhibition of these phenomena. The repression of alfA-lacZ expression was observed in the constructs carrying a mutation in either the first or third repeat, but not in the second repeat, indicating a correlation between plasmid stability, AlfB binding, and repression. It was also demonstrated by the yeast two-hybrid system that AlfA and AlfB interact with each other and among themselves. From these results, it was concluded that AlfB participates in partitioning pBET131 by forming a complex with AlfA and parN, the mode of which is typified by the type II partition mechanism. PMID- 20023010 TI - A novel pheromone quorum-sensing system controls the development of natural competence in Streptococcus thermophilus and Streptococcus salivarius. AB - In streptococcal species, the key step of competence development is the transcriptional induction of comX, which encodes the alternative sigma factor sigma(X), which positively regulates genes necessary for DNA transformation. In Streptococcus species belonging to the mitis and mutans groups, induction of comX relies on the activation of a three-component system consisting of a secreted pheromone, a histidine kinase, and a response regulator. In Streptococcus thermophilus, a species belonging to the salivarius group, the oligopeptide transporter Ami is essential for comX expression under competence-inducing conditions. This suggests a different regulation pathway of competence based on the production and reimportation of a signal peptide. The objective of our work was to identify the main actors involved in the early steps of comX induction in S. thermophilus LMD-9. Using a transcriptomic approach, four highly induced early competence operons were identified. Among them, we found a Rgg-like regulator (Ster_0316) associated with a nonannotated gene encoding a 24-amino-acid hydrophobic peptide (Shp0316). Through genetic deletions, we showed that these two genes are essential for comX induction. Moreover, addition to the medium of synthetic peptides derived from the C-terminal part of Shp0316 restored comX induction and transformation of a Shp0316-deficient strain. These peptides also induced competence in S. thermophilus and Streptococcus salivarius strains that are poorly transformable or not transformable. Altogether, our results show that Ster_0316 and Shp0316, renamed ComRS, are the two members of a novel quorum sensing system responsible for comX induction in species from the salivarius group, which differs from the classical phosphorelay three-component system identified previously in streptococci. PMID- 20023011 TI - Glycine betaine biosynthesized from glycine provides an osmolyte for cell growth and spore germination during osmotic stress in Myxococcus xanthus. AB - Glycine sarcosine methyltransferase (Gsm) and sarcosine dimethylglycine methyltransferase (Sdm) catalyze glycine betaine synthesis from glycine. Disruption of the M. xanthus gsmA (MXAN 7068) or sdmA (MXAN 3190) gene, encoding Gsm or Sdm homologue proteins, respectively, generated mutants that exhibited a longer lag period of growth and delayed spore germination under osmostress. PMID- 20023012 TI - Completed genome sequence of the anaerobic iron-oxidizing bacterium Acidovorax ebreus strain TPSY. AB - Acidovorax ebreus strain TPSY is the first anaerobic nitrate-dependent Fe(II) oxidizer for which there is a completed genome sequence. Preliminary protein annotation revealed an organism optimized for survival in a complex environmental system. Here, we briefly report the completed and annotated genome sequence of strain TPSY. PMID- 20023013 TI - Nitrogenase switch-off and regulation of ammonium assimilation in response to light deprivation in Rhodospirillum rubrum are influenced by the nitrogen source used during growth. AB - Nitrogen fixation and ammonium assimilation in Rhodospirillum rubrum are regulated in response to changes in light availability, and we show that the response in terms of glutamine synthetase activity and P(II) modification is dependent on the nitrogen source used for growth, N(2) or glutamate, although both lead to nitrogenase derepression. PMID- 20023014 TI - Genome-wide identification of targets for the archaeal heat shock regulator phr by cell-free transcription of genomic DNA. AB - The hyperthermophilic archaeon Pyrococcus furiosus grows optimally near 100 degrees C and undergoes a heat shock response at 105 degrees C, mediated at least in part by the heat shock regulator Phr. Genes encoding a small heat shock protein (HSP20) and a member of the AAA(+) ATPase are the only known targets of the regulator, but a genetic mutant of Phr has yet to be characterized. We describe here an alternative approach for the identification of the regulon of Phr based on cell-free transcription of fragmented chromosomal DNA in the presence or absence of the regulator and hybridization of in vitro RNA to P. furiosus whole-genome microarrays. Our results confirmed the phr, the hsp20, and the aaa(+) ATPase genes as targets of Phr and also identified six additional open reading frames, PF0624, PF1042, PF1291, PF1292, PF1488, and PF1616, as Phr responsive genes, which include that encoding di-myo-inositol phosphate synthase. Transcription of the identified novel genes was inhibited by Phr in standard transcription assays, and the novel consensus sequence 5' TTTAnnnACnnnnnGTnAnnAAAA-3' (uppercase letters denote a high conservation of the bases) was inferred from our data as the Phr recognition motif. Mutational evidence for the significance of this sequence as Phr recognition was provided in DNA-binding experiments. PMID- 20023015 TI - Discovery and characterization of cadherin domains in Saccharophagus degradans 2 40. AB - Saccharophagus degradans strain 2-40 is a prominent member of newly discovered group of marine and estuarine bacteria that recycle complex polysaccharides. The S. degradans 2-40 genome codes for 15 extraordinary long polypeptides, ranging from 274 to 1,600 kDa. Five of these contain at least 52 cadherin (CA) and cadherin-like (CADG) domains, the types of which were reported to bind calcium ions and mediate protein/protein interactions in metazoan systems. In order to evaluate adhesive features of these domains, recombinant CA doublet domains (two neighboring domains) from CabC (Sde_3323) and recombinant CADG doublet domains from CabD (Sde_0798) were examined qualitatively and quantitatively for homophilic and heterophilic interactions. In addition, CA and CADG doublet domains were tested for adhesion to the surface of S. degradans 2-40. Results showed obvious homophilic and heterophilic, calcium ion-dependent interactions between CA and CADG doublet domains. Likewise, CA and CADG doublet domains adhered to the S. degradans 2-40 surface of cells that were grown on xylan from birch wood or pectin, respectively, as a sole carbon source. This research shows for the first time that bacterial cadherin homophilic and heterophilic interactions may be similar in their nature to cadherin domains from metazoan lineages. We hypothesize that S. degradans 2-40 cadherin and cadherin-like multiple domains contribute to protein-protein interactions that may mediate cell cell contact in the marine environment. PMID- 20023016 TI - Evidence for rare capsular switching in Streptococcus agalactiae. AB - The polysaccharide capsule is a major antigenic factor in Streptococcus agalactiae (Lancefield group B streptococcus [GBS]). Previous observations suggest that exchange of capsular loci is likely to occur rather frequently in GBS, even though GBS is not known to be naturally transformable. We sought to identify and characterize putative capsular switching events, by means of a combination of phenotypic and genotypic methods, including pulsed-field gel electrophoretic profiling, multilocus sequence typing, and surface protein and pilus gene profiling. We show that capsular switching by horizontal gene transfer is not as frequent as previously suggested. Serotyping errors may be the main reason behind the overestimation of capsule switching, since phenotypic techniques are prone to errors of interpretation. The identified putative capsular transformants involved the acquisition of the entire capsular locus and were not restricted to the serotype-specific central genes, the previously suggested main mechanism underlying capsular switching. Our data, while questioning the frequency of capsular switching, provide clear evidence for in vivo capsular transformation in S. agalactiae, which may be of critical importance in planning future vaccination strategies against this pathogen. PMID- 20023017 TI - CotE binds to CotC and CotU and mediates their interaction during spore coat formation in Bacillus subtilis. AB - CotE is a morphogenic protein that controls the assembly of the coat, the proteinaceous structure that surrounds and protects the spore of Bacillus subtilis. CotE has long been thought to interact with several outer coat components, but such interactions were hypothesized from genetic experiment results and have never been directly demonstrated. To study the interaction of CotE with other coat components, we focused our attention on CotC and CotU, two outer coat proteins known to be under CotE control and to form a heterodimer. We report here the results of pull-down experiments that provide the first direct evidence that CotE contacts other coat components. In addition, coexpression experiments demonstrate that CotE is needed and sufficient to allow formation of the CotC-CotU heterodimer in a heterologous host. PMID- 20023018 TI - Genome diversity of Pseudomonas aeruginosa PAO1 laboratory strains. AB - Pseudomonas aeruginosa PAO1 is the most commonly used strain for research on this ubiquitous and metabolically versatile opportunistic pathogen. Strain PAO1, a derivative of the original Australian PAO isolate, has been distributed worldwide to laboratories and strain collections. Over decades discordant phenotypes of PAO1 sublines have emerged. Taking the existing PAO1-UW genome sequence (named after the University of Washington, which led the sequencing project) as a blueprint, the genome sequences of reference strains MPAO1 and PAO1-DSM (stored at the German Collection for Microorganisms and Cell Cultures [DSMZ]) were resolved by physical mapping and deep short read sequencing-by-synthesis. MPAO1 has been the source of near-saturation libraries of transposon insertion mutants, and PAO1-DSM is identical in its SpeI-DpnI restriction map with the original isolate. The major genomic differences of MPAO1 and PAO1-DSM in comparison to PAO1-UW are the lack of a large inversion, a duplication of a mobile 12-kb prophage region carrying a distinct integrase and protein phosphatases or kinases, deletions of 3 to 1,006 bp in size, and at least 39 single-nucleotide substitutions, 17 of which affect protein sequences. The PAO1 sublines differed in their ability to cope with nutrient limitation and their virulence in an acute murine airway infection model. Subline PAO1-DSM outnumbered the two other sublines in late stationary growth phase. In conclusion, P. aeruginosa PAO1 shows an ongoing microevolution of genotype and phenotype that jeopardizes the reproducibility of research. High-throughput genome resequencing will resolve more cases and could become a proper quality control for strain collections. PMID- 20023019 TI - The Mycobacterium tuberculosis DosR regulon assists in metabolic homeostasis and enables rapid recovery from nonrespiring dormancy. AB - Mycobacterium tuberculosis survives in latently infected individuals, likely in a nonreplicating or dormancy-like state. The M. tuberculosis DosR regulon is a genetic program induced by conditions that inhibit aerobic respiration and prevent bacillus replication. In this study, we used a mutant incapable of DosR regulon induction to investigate the contribution of this regulon to bacterial metabolism during anaerobic dormancy. Our results confirm that the DosR regulon is essential for M. tuberculosis survival during anaerobic dormancy and demonstrate that it is required for metabolic processes that occur upon entry into and throughout the dormant state. Specifically, we showed that regulon mechanisms shift metabolism away from aerobic respiration in the face of dwindling oxygen availability and are required for maintaining energy levels and redox balance as the culture becomes anaerobic. We also demonstrated that the DosR regulon is crucial for rapid resumption of growth once M. tuberculosis exits an anaerobic or nitric oxide-induced nonrespiring state. In summary, the DosR regulon encodes novel metabolic mechanisms essential for M. tuberculosis to survive in the absence of respiration and to successfully transition rapidly between respiring and nonrespiring conditions without loss of viability. PMID- 20023020 TI - Purification of Helicobacter pylori NCTC 11637 cytochrome bc1 and respiration with D-proline as a substrate. AB - Helicobacter pylori is a microaerophilic bacterium associated with gastric inflammation and peptic ulcers. Knowledge of how pathogenic organisms produce energy is important from a therapeutic point of view. We found d-amino acid dehydrogenase-mediated electron transport from d-proline or d-alanine to oxygen via the respiratory chain in H. pylori. Coupling of the electron transport to ATP synthesis was confirmed by using uncoupler reagents. We reconstituted the electron transport chain to demonstrate the electron flow from the d-amino acids to oxygen using the recombinant cytochrome bc(1) complex, cytochrome c-553, and the terminal oxidase cytochrome cbb(3) complex. Upon addition of the recombinant d-amino acid dehydrogenase and d-proline or d-alanine to the reconstituted electron transport system, reduction of cytochrome cbb(3) and oxygen consumption was revealed spectrophotometrically and polarographically, respectively. Among the constituents of H. pylori's electron transport chain, only the cytochrome bc(1) complex had been remained unpurified. Therefore, we cloned and sequenced the H. pylori NCTC 11637 cytochrome bc(1) gene clusters encoding Rieske Fe-S protein, cytochrome b, and cytochrome c(1), with calculated molecular masses of 18 kDa, 47 kDa, and 32 kDa, respectively, and purified the recombinant monomeric protein complex with a molecular mass of 110 kDa by gel filtration. The absorption spectrum of the recombinant cytochrome bc(1) complex showed an alpha peak at 561 nm with a shoulder at 552 nm. PMID- 20023021 TI - The MG1363 and IL1403 laboratory strains of Lactococcus lactis and several dairy strains are diploid. AB - Bacteria are normally haploid, maintaining one copy of their genome in one circular chromosome. We have examined the cell cycle of laboratory strains of Lactococcus lactis, and, to our surprise, we found that some of these strains were born with two complete nonreplicating chromosomes. We determined the cellular content of DNA by flow cytometry and by radioactive labeling of the DNA. These strains thus fulfill the criterion of being diploid. Several dairy strains were also found to be diploid while a nondairy strain and several other dairy strains were haploid in slow-growing culture. The diploid and haploid strains differed in their sensitivity toward UV light, in their cell size, and in their D period, the period between termination of DNA replication and cell division. PMID- 20023022 TI - The El Tor biotype of Vibrio cholerae exhibits a growth advantage in the stationary phase in mixed cultures with the classical biotype. AB - Vibrio cholerae strains of the O1 serogroup that typically cause epidemic cholera can be classified into two biotypes, classical and El Tor. The El Tor biotype emerged in 1961 and subsequently displaced the classical biotype as a cause of cholera throughout the world. In this study we demonstrate that when strains of the El Tor and classical biotypes were cocultured in standard LB medium, the El Tor strains clearly had a competitive growth advantage over the classical biotype starting from the late stationary phase and could eventually take over the population. The classical biotype produces extracellular protease(s) in the stationary phase, and the amounts of amino acids and small peptides in the late stationary and death phase culture filtrates of the classical biotype were higher than those in the corresponding culture filtrates of the El Tor biotype. The El Tor biotype cells could utilize the amino acids more efficiently than the classical biotype under the alkaline pH of the stationary phase cultures but not in medium buffered to neutral pH. The growth advantage of the El Tor biotype was also observed in vivo using the ligated rabbit ileal loop and infant mouse animal models. PMID- 20023023 TI - Plasmid segregation: birds of a feather try not to flock together. PMID- 20023024 TI - The nonphosphorylative Entner-Doudoroff pathway in the thermoacidophilic euryarchaeon Picrophilus torridus involves a novel 2-keto-3-deoxygluconate- specific aldolase. AB - The pathway of glucose degradation in the thermoacidophilic euryarchaeon Picrophilus torridus has been studied by in vivo labeling experiments and enzyme analyses. After growth of P. torridus in the presence of [1-(13)C]- and [3 (13)C]glucose, the label was found only in the C-1 and C-3 positions, respectively, of the proteinogenic amino acid alanine, indicating the exclusive operation of an Entner-Doudoroff (ED)-type pathway in vivo. Cell extracts of P. torridus contained all enzyme activities of a nonphosphorylative ED pathway, which were not induced by glucose. Two key enzymes, gluconate dehydratase (GAD) and a novel 2-keto-3-deoxygluconate (KDG)-specific aldolase (KDGA), were characterized. GAD is a homooctamer of 44-kDa subunits, encoded by Pto0485. KDG aldolase, KDGA, is a homotetramer of 32-kDa subunits. This enzyme was highly specific for KDG with up to 2,000-fold-higher catalytic efficiency compared to 2 keto-3-deoxy-6-phosphogluconate (KDPG) and thus differs from the bifunctional KDG/KDPG aldolase, KD(P)GA of crenarchaea catalyzing the conversion of both KDG and KDPG with a preference for KDPG. The KDGA-encoding gene, kdgA, was identified by matrix-assisted laser desorption ionization-time of flight (MALDI-TOF) mass spectrometry (MS) as Pto1279, and the correct translation start codon, an ATG 24 bp upstream of the annotated start codon of Pto1279, was determined by N-terminal amino acid analysis. The kdgA gene was functionally overexpressed in Escherichia coli. Phylogenetic analysis revealed that KDGA is only distantly related to KD(P)GA, both enzymes forming separate families within the dihydrodipicolinate synthase superfamily. From the data we conclude that P. torridus degrades glucose via a strictly nonphosphorylative ED pathway with a novel KDG-specific aldolase, thus excluding the operation of the branched ED pathway involving a bifunctional KD(P)GA as a key enzyme. PMID- 20023025 TI - Characterization of a glutamate transporter operon, glnQHMP, in Streptococcus mutans and its role in acid tolerance. AB - Glutamate contributes to the acid tolerance response (ATR) of many Gram-negative and Gram-positive bacteria, but its role in the ATR of the oral bacterium Streptococcus mutans is unknown. This study describes the discovery and characterization of a glutamate transporter operon designated glnQHMP (Smu.1519 to Smu.1522) and investigates its potential role in acid tolerance. Deletion of glnQHMP resulted in a 95% reduction in transport of radiolabeled glutamate compared to the wild-type UA159 strain. The addition of glutamate to metabolizing UA159 cells resulted in an increased production of acidic end products, whereas the glnQHMP mutant produced less lactic acid than UA159, suggesting a link between glutamate metabolism and acid production and possible acid tolerance. To investigate this possibility, we conducted a microarray analysis with glutamate and under pH 5.5 and pH 7.5 conditions which showed that expression of the glnQHMP operon was downregulated by both glutamate and mild acid. We also measured the growth kinetics of UA159 and its glnQHMP-negative derivative at pH 5.5 and found that the mutant doubled at a much slower rate than the parent strain but survived at pH 3.5 significantly better than the wild type. Taken together, these findings support the involvement of the glutamate transporter operon glnQHMP in the acid tolerance response in S. mutans. PMID- 20023026 TI - Characterization of a gene family of outer membrane proteins (ropB) in Rhizobium leguminosarum bv. viciae VF39SM and the role of the sensor kinase ChvG in their regulation. AB - The outer membrane of Gram-negative bacteria represents the interface between the bacterium and its external environment. It has a critical role as a protective barrier against harmful substances and is also important in host-bacteria interactions representing the initial physical point of contact between the host cell and bacterial cell. RopB is a previously identified outer membrane protein from Rhizobium leguminosarum bv. viciae that is present in free-living cells but absent in bacteroids (H. P. Roest, I. H. Mulders, C. A. Wijffelman, and B. J. Lugtenberg, Mol. Plant Microbe Interact. 8:576-583, 1995). The functions of RopB and the molecular mechanisms of ropB gene regulation have remained unknown. We identified and cloned ropB and two homologs (ropB2 and ropB3) from the R. leguminosarum VF39SM genome. Reporter gene fusions indicated that the expression of ropB was 8-fold higher when cells were grown in complex media than when they were grown in minimal media, while ropB3 expression was constitutively expressed at low levels in both complex and minimal media. Expression of ropB2 was negligible under all conditions tested. The use of minimal media supplemented with various sources of peptides resulted in a 5-fold increase in ropB expression. An increase in ropB expression in the presence of peptides was not observed in a chvG mutant background, indicating a role for the sensor kinase in regulating ropB expression. Each member of the ropB gene family was mutated using insertional mutagenesis, and the mutants were assayed for susceptibility to antimicrobial agents and symbiotic phenotypes. All mutants formed effective nodules on pea plants, and gene expression for each rop gene in bacteroids was negligible. The functions of ropB2 and ropB3 remain cryptic, while the ropB mutant had an increased sensitivity to detergents, hydrophobic antibiotics, and weak organic acids, suggesting a role for RopB in outer membrane stability. PMID- 20023027 TI - The genome of the amoeba symbiont "Candidatus Amoebophilus asiaticus" reveals common mechanisms for host cell interaction among amoeba-associated bacteria. AB - Protozoa play host for many intracellular bacteria and are important for the adaptation of pathogenic bacteria to eukaryotic cells. We analyzed the genome sequence of "Candidatus Amoebophilus asiaticus," an obligate intracellular amoeba symbiont belonging to the Bacteroidetes. The genome has a size of 1.89 Mbp, encodes 1,557 proteins, and shows massive proliferation of IS elements (24% of all genes), although the genome seems to be evolutionarily relatively stable. The genome does not encode pathways for de novo biosynthesis of cofactors, nucleotides, and almost all amino acids. "Ca. Amoebophilus asiaticus" encodes a variety of proteins with predicted importance for host cell interaction; in particular, an arsenal of proteins with eukaryotic domains, including ankyrin-, TPR/SEL1-, and leucine-rich repeats, which is hitherto unmatched among prokaryotes, is remarkable. Unexpectedly, 26 proteins that can interfere with the host ubiquitin system were identified in the genome. These proteins include F- and U-box domain proteins and two ubiquitin-specific proteases of the CA clan C19 family, representing the first prokaryotic members of this protein family. Consequently, interference with the host ubiquitin system is an important host cell interaction mechanism of "Ca. Amoebophilus asiaticus". More generally, we show that the eukaryotic domains identified in "Ca. Amoebophilus asiaticus" are also significantly enriched in the genomes of other amoeba-associated bacteria (including chlamydiae, Legionella pneumophila, Rickettsia bellii, Francisella tularensis, and Mycobacterium avium). This indicates that phylogenetically and ecologically diverse bacteria which thrive inside amoebae exploit common mechanisms for interaction with their hosts, and it provides further evidence for the role of amoebae as training grounds for bacterial pathogens of humans. PMID- 20023028 TI - Escherichia coli RNase R has dual activities, helicase and RNase. AB - In Escherichia coli, the cold shock response occurs when there is a temperature downshift from 37 degrees C to 15 degrees C, and this response is characterized by induction of several cold shock proteins, including the DEAD-box helicase CsdA, during the acclimation phase. CsdA is involved in a variety of cellular processes. Our previous studies showed that the helicase activity of CsdA is critical for its function in cold shock acclimation of cells and that the only proteins that were able to complement its function were another helicase, RhlE, an RNA chaperone, CspA, and a cold-inducible exoribonuclease, RNase R. Interestingly, other major 3'-to-5' processing exoribonucleases of E. coli, such as polynucleotide phosphorylase and RNase II, cannot complement the cold shock function of CsdA. Here we carried out a domain analysis of RNase R and showed that this protein has two distinct activities, RNase and helicase, which are independent of each other and are due to different domains. Mutant RNase R proteins that lack the RNase activity but exhibit the helicase activity were able to complement the cold shock function of CsdA, suggesting that only the helicase activity of RNase R is essential for complementation of the cold shock function of CsdA. We also observed that in vivo deletion of the two cold shock domains resulted in a loss of the ability of RNase R to complement the cold shock function of CsdA. We further demonstrated that RNase R exhibits helicase activity in vitro independent of its RNase activity. Our results shed light on the unique properties of RNase R and how it is distinct from other exoribonucleases in E. coli. PMID- 20023029 TI - Manipulating each MreB of Bdellovibrio bacteriovorus gives diverse morphological and predatory phenotypes. AB - We studied the two mreB genes, encoding actinlike cytoskeletal elements, in the predatory bacterium Bdellovibrio bacteriovorus. This bacterium enters and replicates within other Gram-negative bacteria by attack-phase Bdellovibrio squeezing through prey outer membrane, residing and growing filamentously in the prey periplasm forming an infective "bdelloplast," and septating after 4 h, once the prey contents are consumed. This lifestyle brings challenges to the Bdellovibrio cytoskeleton. Both mreB genes were essential for viable predatory growth, but C-terminal green fluorescent protein tagging each separately with monomeric teal-fluorescent protein (mTFP) gave two strains with phenotypic changes at different stages in predatory growth and development. MreB1-mTFP cells arrested growth early in bdelloplast formation, despite successful degradation of prey nucleoid. A large population of stalled bdelloplasts formed in predatory cultures and predation proceeded very slowly. A small proportion of bdelloplasts lysed after several days, liberating MreB1-mTFP attack-phase cells of wild-type morphology; this process was aided by subinhibitory concentrations of an MreB specific inhibitor, A22. MreB2-mTFP, in contrast, was predatory at an almost wild type rate but yielded attack-phase cells with diverse morphologies, including spherical, elongated, and branched, the first time such phenotypes have been described. Wild-type predatory rates were seen for all but spherical morphotypes, and septation of elongated morphotypes was achieved by the addition of A22. PMID- 20023030 TI - Characterization of CamH from Methanosarcina thermophila, founding member of a subclass of the {gamma} class of carbonic anhydrases. AB - The homotrimeric enzyme Mt-Cam from Methanosarcina thermophila is the archetype of the gamma class of carbonic anhydrases. A search of databases queried with Mt Cam revealed that a majority of the homologs comprise a putative subclass (CamH) in which there is major conservation of all of the residues essential for the archetype Mt-Cam except Glu62 and an acidic loop containing the essential proton shuttle residue Glu84. The CamH homolog from M. thermophila (Mt-CamH) was overproduced in Escherichia coli and characterized to validate its activity and initiate an investigation of the CamH subclass. The Mt-CamH homotrimer purified from E. coli cultured with supplemental zinc (Zn-Mt-CamH) contained 0.71 zinc and 0.15 iron per monomer and had k(cat) and k(cat)/K(m) values that were substantially lower than those for the zinc form of Mt-Cam (Zn-Mt-Cam). Mt-CamH purified from E. coli cultured with supplemental iron (Fe-Mt-CamH) was also a trimer containing 0.15 iron per monomer and only a trace amount of zinc and had an effective k(cat) (k(cat)(eff)) value normalized for iron that was 6-fold less than that for the iron form of Mt-Cam, whereas the k(cat)/K(m)(eff) was similar to that for Fe-Mt-Cam. Addition of 50 mM imidazole to the assay buffer increased the k(cat)(eff) of Fe-Mt-CamH more than 4-fold. Fe-Mt-CamH lost activity when it was exposed to air or 3% H(2)O(2), which supports the hypothesis that Fe(2+) has a role in the active site. The k(cat) for Fe-Mt-CamH was dependent on the concentration of buffer in a way that indicates that it acts as a second substrate in a "ping-pong" mechanism accepting a proton. The k(cat)/K(m) was not dependent on the buffer, consistent with the mechanism for all carbonic anhydrases in which the interconversion of CO(2) and HCO(3)(-) is separate from intermolecular proton transfer. PMID- 20023031 TI - Nucleases encoded by the integrated elements CJIE2 and CJIE4 inhibit natural transformation of Campylobacter jejuni. AB - The species Campylobacter jejuni is naturally competent for DNA uptake; nevertheless, nonnaturally transformable strains do exist. For a subset of strains we previously showed that a periplasmic DNase, encoded by dns, inhibits natural transformation in C. jejuni. In the present study, genetic factors coding for DNase activity in the absence of dns were identified. DNA arrays indicated that nonnaturally transformable dns-negative strains contain putative DNA/RNA nonspecific endonucleases encoded by CJE0566 and CJE1441 of strain RM1221. These genes are located on C. jejuni integrated elements 2 and 4. Expression of CJE0566 and CJE1441 from strain RM1221 and a homologous gene from strain 07479 in DNase negative Escherichia coli and C. jejuni strains indicated that these genes code for DNases. Genetic transfer of the genes to a naturally transformable C. jejuni strain resulted in a decreased efficiency of natural transformation. Modeling suggests that the C. jejuni DNases belong to the Serratia nuclease family. Overall, the data indicate that the acquisition of prophage-encoded DNA/RNA nonspecific endonucleases inhibits the natural transformability of C. jejuni through hydrolysis of DNA. PMID- 20023032 TI - Production of the Bsa lantibiotic by community-acquired Staphylococcus aureus strains. AB - Lantibiotics are antimicrobial peptides that have been the focus of much attention in recent years with a view to clinical, veterinary, and food applications. Although many lantibiotics are produced by food-grade bacteria or bacteria generally regarded as safe, some lantibiotics are produced by pathogens and, rather than contributing to food safety and/or health, add to the virulence potential of the producing strains. Indeed, genome sequencing has revealed the presence of genes apparently encoding a lantibiotic, designated Bsa (bacteriocin of Staphylococcus aureus), among clinical isolates of S. aureus and those associated with community-acquired methicillin-resistant S. aureus (MRSA) infections in particular. Here, we establish for the first time, through a combination of reverse genetics, mass spectrometry, and mutagenesis, that these genes encode a functional lantibiotic. We also reveal that Bsa is identical to the previously identified bacteriocin staphylococcin Au-26, produced by an S. aureus strain of vaginal origin. Our examination of MRSA isolates that produce the Panton-Valentine leukocidin demonstrates that many community-acquired S. aureus strains, and representatives of ST8 and ST80 in particular, are producers of Bsa. While possession of Bsa immunity genes does not significantly enhance resistance to the related lantibiotic gallidermin, the broad antimicrobial spectrum of Bsa strongly indicates that production of this bacteriocin confers a competitive ecological advantage on community-acquired S. aureus. PMID- 20023033 TI - Deletion of the ftsZ-like gene results in the production of superparamagnetic magnetite magnetosomes in Magnetospirillum gryphiswaldense. AB - Magnetotactic bacteria (MTB) synthesize unique organelles termed "magnetosomes," which are membrane-enclosed structures containing crystals of magnetite or greigite. Magnetosomes form a chain around MamK cytoskeletal filaments and provide the basis for the ability of MTB to navigate along geomagnetic field lines in order to find optimal microaerobic habitats. Genomes of species of the MTB genus Magnetospirillum, in addition to a gene encoding the tubulin-like FtsZ protein (involved in cell division), contain a second gene termed "ftsZ-like," whose function is unknown. In the present study, we found that the ftsZ-like gene of Magnetospirillum gryphiswaldense strain MSR-1 belongs to a 4.9-kb mamXY polycistronic transcription unit. We then purified the recombinant FtsZ-like protein to homogeneity. The FtsZ-like protein efficiently hydrolyzed ATP and GTP, with ATPase and GTPase activity levels of 2.17 and 5.56 mumol phosphorus per mol protein per min, respectively. The FtsZ-like protein underwent GTP-dependent polymerization into long filamentous bundles in vitro. To determine the role of the ftsZ-like gene, we constructed a ftsZ-like mutant (DeltaftsZ-like mutant) and its complementation strain (DeltaftsZ-like_C strain). Growth of DeltaftsZ-like cells was similar to that of the wild type, indicating that the DeltaftsZ-like gene is not involved in cell division. Transmission electron microscopic observations indicated that the DeltaftsZ-like cells, in comparison to wild-type cells, produced smaller magnetosomes, with poorly defined morphology and irregular alignment, including large gaps. Magnetic analyses showed that DeltaftsZ-like produced mainly superparamagnetic (SP) magnetite particles, whereas wild-type and DeltaftsZ-like_C cells produced mainly single-domain (SD) particles. Our findings suggest that the FtsZ-like protein is required for synthesis of SD particles and magnetosomes in M. gryphiswaldense. PMID- 20023034 TI - Bacterial ammeline metabolism via guanine deaminase. AB - Melamine toxicity in mammals has been attributed to the blockage of kidney tubules by insoluble complexes of melamine with cyanuric acid or uric acid. Bacteria metabolize melamine via three consecutive deamination reactions to generate cyanuric acid. The second deamination reaction, in which ammeline is the substrate, is common to many bacteria, but the genes and enzymes responsible have not been previously identified. Here, we combined bioinformatics and experimental data to identify guanine deaminase as the enzyme responsible for this biotransformation. The ammeline degradation phenotype was demonstrated in wild type Escherichia coli and Pseudomonas strains, including E. coli K12 and Pseudomonas putida KT2440. Bioinformatics analysis of these and other genomes led to the hypothesis that the ammeline deaminating enzyme was guanine deaminase. An E. coli guanine deaminase deletion mutant was deficient in ammeline deaminase activity, supporting the role of guanine deaminase in this reaction. Two guanine deaminases from disparate sources (Bradyrhizobium japonicum USDA 110 and Homo sapiens) that had available X-ray structures were purified to homogeneity and shown to catalyze ammeline deamination at rates sufficient to support bacterial growth on ammeline as a sole nitrogen source. In silico models of guanine deaminase active sites showed that ammeline could bind to guanine deaminase in a similar orientation to guanine, with a favorable docking score. Other members of the amidohydrolase superfamily that are not guanine deaminases were assayed in vitro, and none had substantial ammeline deaminase activity. The present study indicated that widespread guanine deaminases have a promiscuous activity allowing them to catalyze a key reaction in the bacterial transformation of melamine to cyanuric acid and potentially contribute to the toxicity of melamine. PMID- 20023035 TI - MreB drives de novo rod morphogenesis in Caulobacter crescentus via remodeling of the cell wall. AB - MreB, the bacterial actin-like cytoskeleton, is required for the rod morphology of many bacterial species. Disruption of MreB function results in loss of rod morphology and cell rounding. Here, we show that the widely used MreB inhibitor A22 causes MreB-independent growth inhibition that varies with the drug concentration, culture medium conditions, and bacterial species tested. MP265, an A22 structural analog, is less toxic than A22 for growth yet equally efficient for disrupting the MreB cytoskeleton. The action of A22 and MP265 is enhanced by basic pH of the culture medium. Using this knowledge and the rapid reversibility of drug action, we examined the restoration of rod shape in lemon-shaped Caulobacter crescentus cells pretreated with MP265 or A22 under nontoxic conditions. We found that reversible restoration of MreB function after drug removal causes extensive morphological changes including a remarkable cell thinning accompanied with elongation, cell branching, and shedding of outer membrane vesicles. We also thoroughly characterized the composition of C. crescentus peptidoglycan by high-performance liquid chromatography and mass spectrometry and showed that MreB disruption and recovery of rod shape following restoration of MreB function are accompanied by considerable changes in composition. Our results provide insight into MreB function in peptidoglycan remodeling and rod shape morphogenesis and suggest that MreB promotes the transglycosylase activity of penicillin-binding proteins. PMID- 20023036 TI - Rhizobium leguminosarum hupE encodes a nickel transporter required for hydrogenase activity. AB - Synthesis of the hydrogen uptake (Hup) system in Rhizobium leguminosarum bv. viciae requires the function of an 18-gene cluster (hupSLCDEFGHIJK-hypABFCDEX). Among them, the hupE gene encodes a protein showing six transmembrane domains for which a potential role as a nickel permease has been proposed. In this paper, we further characterize the nickel transport capacity of HupE and that of the translated product of hupE2, a hydrogenase-unlinked gene identified in the R. leguminosarum genome. HupE2 is a potential membrane protein that shows 48% amino acid sequence identity with HupE. Expression of both genes in the Escherichia coli nikABCDE mutant strain HYD723 restored hydrogenase activity and nickel transport. However, nickel transport assays revealed that HupE and HupE2 displayed different levels of nickel uptake. Site-directed mutagenesis of histidine residues in HupE revealed two motifs (HX(5)DH and FHGX[AV]HGXE) that are required for HupE functionality. An R. leguminosarum double mutant, SPF22A (hupE hupE2), exhibited reduced levels of hydrogenase activity in free-living cells, and this phenotype was complemented by nickel supplementation. Low levels of symbiotic hydrogenase activity were also observed in SPF22A bacteroid cells from lentil (Lens culinaris L.) root nodules but not in pea (Pisum sativum L.) bacteroids. Moreover, heterologous expression of the R. leguminosarum hup system in bacteroid cells of Rhizobium tropici and Mesorhizobium loti displayed reduced levels of hydrogen uptake in the absence of hupE. These data support the role of R. leguminosarum HupE as a nickel permease required for hydrogen uptake under both free-living and symbiotic conditions. PMID- 20023039 TI - MEF2 transcriptional activity maintains mitochondrial adaptation in cardiac pressure overload. AB - AIMS: The transcription factor MEF2 is a downstream target for several hypertrophic signalling pathways in the heart, suggesting that MEF2 may act as a valuable therapeutic target in the treatment of heart failure. METHODS AND RESULTS: In this study, we investigated the potential benefits of overall MEF2 inhibition in a mouse model of chronic pressure overloading, by subjecting transgenic mice expressing a dominant negative form of MEF2 (DN-MEF2 Tg) in the heart, to transverse aortic constriction (TAC). Histological analysis revealed no major differences in cardiac remodelling between DN-MEF2 Tg and control mice after TAC. Surprisingly, echocardiographic analysis revealed that DN-MEF2 Tg mice had a decrease in cardiac function compared with control animals. Analysis of the mitochondrial respiratory chain showed that DN-MEF2 Tg mice displayed lower expression of NADH dehydrogenase subunit 6 (ND6), part of mitochondrial Complex I. The reduced expression of ND6 in DN-MEF2 Tg mice after pressure overload correlated with an increase in cell death secondary to overproduction of reactive oxygen species (ROS). CONCLUSION: Our data suggest that MEF2 transcriptional activity is required for mitochondrial function and its inhibition predisposes the heart to impaired mitochondrial function, overproduction of ROS, enhanced cell death, and cardiac dysfunction, following pressure overload. PMID- 20023040 TI - L41Q polymorphism of the G protein coupled receptor kinase 5 is associated with left ventricular apical ballooning syndrome. PMID- 20023041 TI - Heart failure registry: a valuable tool for improving the management of patients with heart failure. AB - AIMS: Guidelines on how to diagnose and treat patients with heart failure (HF) are published regularly. However, many patients do not fulfil the diagnostic criteria and are not treated with recommended drugs. The Swedish Heart Failure Registry (S-HFR) is an instrument which may help to optimize the handling of HF patients. METHODS AND RESULTS: The S-HFR is an Internet-based registry in which participating centres (units) can record details of their HF patients directly online and transfer data from standardized forms or from computerized patient documentation. Up to December 2007, 16,117 patients from 78 units had been included in the S-HFR. Of these, 10,229 patients had been followed for at least 1 year, and 2133 deaths were recorded. Online reports from the registry showed that electrocardiograms were available for 97% of the patients. Sinus rhythm was found in 51% of patients and atrial fibrillation in 38%. Echocardiography was performed in 83% of the patients. Overall, 77% of patients were treated with angiotensin converting enzyme inhibitors or angiotensin II receptor blockers, 80% were on beta-blockers, 34% on aldosterone antagonists, and 83% on diuretics. CONCLUSION: The S-HFR is a valuable tool for improving the management of patients with HF, since it enables participating centres to focus on their own potential for improving diagnoses and medical treatment, through the online reports provided. PMID- 20023042 TI - Acute kidney injury and outcomes in acute decompensated heart failure: evaluation of the RIFLE criteria in an acutely ill heart failure population. AB - AIMS: The clinical course including the outcome of acute decompensated heart failure (ADHF) correlates with renal dysfunction, but the evaluation of renal function has not yet been standardized. We therefore investigated the relationship between the prognosis of ADHF and acute kidney injury (AKI) evaluated using the risk, injury, failure, loss, end stage (RIFLE) criteria. METHODS AND RESULTS: This study assessed 376 consecutive patients with ADHF admitted to the intensive care unit (ICU) (mean age 71.6 years; 238 male). The underlying aetiology was ischaemic heart disease, hypertensive heart disease, cardiomyopathy, valvular diseases, and 'other' in 124, 70, 60, 107, and 15 patients, respectively. We defined AKI according to the RIFLE criteria, and the most severe RIFLE classifications during hospitalization were adopted to assess patient outcomes. The in-hospital mortality was significantly higher among patients with AKI (29 of 275; 10.5%) than in those without AKI (1 of 101; 1.0%, P = 0.0010). Both ICU and hospital stays were longer for patients with AKI (8.8 +/- 15.4 vs. 48.6 +/- 47.6 days), than for patients without (5.0 +/- 2.8 vs. 25.7 +/- 16.8 days, P < 0.05 and P < 0.001). CONCLUSION: Acute kidney injury evaluated by the RIFLE criteria was associated with a poorer outcome for patients with ADHF. PMID- 20023043 TI - Differential prognostic effect of systolic blood pressure on mortality according to left-ventricular function in patients with acute heart failure. AB - AIMS: To evaluate the relationship between systolic blood pressure (SBP) and long term mortality in patients with acute heart failure (AHF) stratified by ejection fraction (LVEF): reduced (< or =40%) vs. preserved (> or =50%). METHODS AND RESULTS: We studied 1049 consecutive patients admitted with AHF. Systolic blood pressure was determined in the emergency department. Left-ventricular ejection fraction was categorized as < or =40% (n = 288), 41-49% (n = 174), or > or =50% (n = 587). Cox regression analysis was used for multivariable analysis. Mean age and SBP were 73 +/- 11 years and 150 +/- 36 mmHg, respectively. During a median follow-up of 18 months, 290 deaths (33.1%) were identified. Higher SBP was associated with lower mortality. In multivariable analysis, a differential effect of SBP across LVEF status was documented (P-value for interaction = 0.036). In linear models, SBP was shown to be inversely related with mortality in both groups (per 10 mmHg decrease): HR((LVEF > or = 50%)): 1.06, CI 95% = 1.01-1.11; P = 0.016, and HR((LVEF < or = 40%)): 1.16, 95% CI = 1.08-1.25; P < 0.001). When SBP was modelled with restrictive cubic splines, an inverse and almost linear relationship with mortality was shown in patients with LVEF < or =40% (P < 0.001), whereas in patients with LVEF > or =50%, SBP followed a J-shape curve. CONCLUSION: In patients with AHF, SBP showed a differential prognostic effect on mortality according to LVEF status; when LVEF was < or =40%, SBP was linearly and inversely associated with mortality. Conversely, in patients with LVEF > or =50% this relationship showed a J-shape pattern. PMID- 20023044 TI - Diagnostic performance of multisequential cardiac magnetic resonance imaging in acute cardiac allograft rejection. AB - AIMS: We evaluated cardiac magnetic resonance imaging (CMR) as a non-invasive test for cardiac allograft rejection. METHODS AND RESULTS: We performed CMR on 50 heart-transplant recipients. Acute rejection was confirmed in 11 cases by endomyocardial biopsy (EMB) and presumed in 8 cases with a recent fall in left ventricular ejection fraction (LVEF) not attributable to coronary allograft vasculopathy. Control patients had both normal LVEF and no significant rejection on EMB. Cardiac magnetic resonance imaging evaluated myocardial function, oedema, and early and late post-Gadolinium-DTPA contrast enhancement. Patients with confirmed rejection demonstrated elevated early relative myocardial contrast enhancement (4.1 +/- 0.3 vs. 2.8 +/- 0.2, P < 0.001) and a trend to higher oedema suggested by higher relative myocardial intensity on T(2)-weighted imaging compared to controls (2.1 +/- 0.1 vs. 1.7 +/- 0.1, P = 0.1). With rejection defined as increased early contrast enhancement or myocardial oedema, the sensitivity and specificity of CMR compared with EMB were 100 and 73%, respectively. Eight patients with presumed rejection also had elevated early myocardial contrast enhancement compared with controls, (8.7 +/- 1.9 vs. 2.8 +/- 0.2, P < 0.05), which reduced following increased immunosuppression (8.7 +/- 1.9 vs. 4.6 +/- 1.2, P < 0.05). In these patients LVEF improved following increased immunosuppression (32 +/- 5 vs. 46 +/- 5%, P < 0.05). CONCLUSION: Cardiac magnetic resonance imaging is a promising modality for non-invasive detection of cardiac allograft rejection. PMID- 20023045 TI - Integrating functional and anatomical information to guide cardiac resynchronization therapy. PMID- 20023046 TI - Impact of gender on benefits of exercise training on sympathetic nerve activity and muscle blood flow in heart failure. AB - AIMS: We compared the effects of exercise training on neurovascular control and functional capacity in men and women with chronic heart failure (HF). METHODS AND RESULTS: Forty consecutive HF outpatients from the Heart Institute, University of Sao Paulo, Brazil were divided into the following four groups matched by age: men exercise-trained (n = 12), men untrained (n = 10), women exercise-trained (n = 9), women untrained (n = 9). Maximal exercise capacity was determined from a maximal progressive exercise test on a cycle ergometer. Forearm blood flow was measured by venous occlusion plethysmography. Muscle sympathetic nerve activity (MSNA) was recorded directly using the technique of microneurography. There were no differences between groups in any baseline parameters. Exercise training produced a similar reduction in resting MSNA (P = 0.000002) and forearm vascular resistance (P = 0.0003), in men and women with HF. Peak VO(2) was similarly increased in men and women with HF (P = 0.0003) and VE/VCO(2) slope was significantly decreased in men and women with HF (P = 0.0007). There were no significant changes in left-ventricular ejection fraction in men and women with HF. CONCLUSION: The benefits of exercise training on neurovascular control and functional capacity in patients with HF are independent of gender. PMID- 20023048 TI - Caveat medicus! Pulmonary hypertension in the elderly: a word of caution. AB - Heart failure with preserved ejection fraction (HFpEF) is a common but under recognized cause of pulmonary hypertension (PH), particularly among the elderly. While elevated pulmonary artery pressures are clearly associated with increased mortality in HFpEF, it remains unknown whether or how pulmonary vascular disease in HFpEF should be treated. This case emphasizes the importance of maintaining a high index of suspicion for HFpEF in elderly patients presenting with PH and illustrates potential hazards associated with selective pulmonary vasodilation in this growing population. PMID- 20023047 TI - An economic evaluation of rosuvastatin treatment in systolic heart failure: evidence from the CORONA trial. AB - AIMS: To estimate the cost-effectiveness of 10 mg rosuvastatin daily for older patients with systolic heart failure in the Controlled Rosuvastatin Multinational Study in Heart Failure (CORONA) trial. METHODS AND RESULTS: This within trial analysis of CORONA used major cardiovascular (CV) events as the outcome measure. Resource use was valued and the costs of hospitalizations, procedures, and statin use compared. Cost-effectiveness was estimated as cost per major CV event avoided. There were significantly fewer major CV events in the rosuvastatin group compared with the placebo group (1.04 vs. 1.20 per patient; difference 0.164; 95% CI: 0.075-0.254, P < 0.001). The average cost of CV hospitalizations and procedures was significantly lower for those receiving rosuvastatin ( pound1531 vs. pound1769; difference pound238; 95% CI: pound73-403, P = 0.005); the additional cost of the statin resulted in significantly higher total costs for the rosuvastatin group ( pound1769 vs. pound2072; difference pound303; 95% CI: pound138-468, P < 0.001). Overall, rosuvastatin was found to cost pound1840 (95% CI: pound562-6028) per major CV event avoided. CONCLUSION: This economic analysis showed that a significant reduction in major CV events with rosuvastatin led to significantly reduced costs of CV hospitalizations and procedures. The reduction in associated costs for major CV events was found to offset partially (by 44%) the cost of rosuvastatin treatment in patients with systolic heart failure. PMID- 20023050 TI - Developing a quality screening colonoscopy referral system in primary care practice: a report from the national colorectal cancer roundtable. AB - The use of colonoscopy in colorectal cancer (CRC) screening has increased substantially in recent years. Media messages and changes in insurance reimbursement, as well as new screening guidelines from the American Cancer Society and the US Preventive Services Task Force, have contributed to this increase. Primary care providers (PCPs) are frequently responsible for making the recommendation and referral for screening. The process of successfully referring a patient for screening colonoscopy can be cumbersome and requires a coordinated effort between the PCP and the endoscopist. In recognition of the potential complexity of this process, the National Colorectal Cancer Roundtable has issued a report to describe the components of a quality screening colonoscopy referral system in primary care practice. The elements of a quality program include an optimal scheduling and referral system, the appropriate patient preparation information, consistent reporting and follow-up systems, and a detailed approach to dealing with special situations. PMID- 20023051 TI - Investigation of the involvement of P-glycoprotein and multidrug resistance associated protein 2 in the efflux of ximelagatran and its metabolites by using short hairpin RNA knockdown in Caco-2 cells. AB - Liver and bile secretion can be an important first-pass and clearance route for drug compounds and also the site of several drug-drug interactions. In the clinical program for ximelagatran development, an unexpected effect of erythromycin on the pharmacokinetics of the direct thrombin inhibitor ximelagatran and its metabolites was detected. This interaction was believed to be mediated by inhibition of drug transporters, which normally extrude the drug into the bile. Previous Caco-2 cell experiments indicated the involvement of an active efflux mechanism for ximelagatran, hydroxy-melagatran, and melagatran possibly mediated by P-glycoprotein (P-gp). However, the inhibitors used may not have been specific enough and the possibility that transporters other than P-gp were important in the Caco-2 cell assay cannot be excluded. In this study we used RNA interference, a post-transcriptional gene silencing mechanism in which mRNA is degraded in a sequence-specific manner, to specifically knock down P-gp or multidrug resistance-associated protein 2 (MRP2) transporters in Caco-2 cells. The data obtained from bidirectional transport studies in these cells indicate a clear involvement of P-gp but not of MRP2 in the transport of ximelagatran, hydroxy-melagatran, and melagatran across the apical cell membrane. The present study shows that short hairpin RNA Caco-2 cells are a valuable tool to investigate the contribution of specific transporters in the transcellular transport of drug molecules and to predict potential sites of pharmacokinetic interactions. The results also suggest that inhibition of hepatic P-gp is involved in the erythromycin-ximelagatran interaction seen in clinical studies. PMID- 20023052 TI - Acute urinary retention due to viral coinfections (HIV, HBV, VZV). AB - We report a case of acute urinary retention with meningoradiculoneuritis resulting from coinfection with varicella-zoster virus (VZV), HIV, and hepatitis B virus (HBV). The case is discussed in the context of other neurological syndromes associated with VZV and HIV infections, and of other viral causes of acute retention of urine. PMID- 20023053 TI - Cupriavidus pampae sp. nov., a novel herbicide-degrading bacterium isolated from agricultural soil. AB - A bacterial consortium able to degrade the herbicide 4-(2,4-dichlorophenoxy) butyric acid (2,4-DB) was obtained from an agricultural soil of the Argentinean Humid Pampa region which has a history of long-term herbicide use. Four bacterial strains were isolated from the consortium and identified as members of the genera Cupriavidus, Labrys and Pseudomonas. A polyphasic systematic analysis was carried out on strain CPDB6(T), the member of the 2,4-DB-degrading consortium able to degrade 2,4-DB as a sole carbon and energy source. The Gram-negative, rod-shaped, motile, non-sporulating, non-fermenting bacterium was shown to belong to the genus Cupriavidus on the basis of 16S rRNA gene sequence analyses. Strain CPDB6(T) did not reduce nitrate, which differentiated it from the type species of the genus, Cupriavidus necator; it did not grow in 0.5-4.5 % NaCl, although most species of Cupriavidus are able to grow at NaCl concentrations as high as 1.5 %; and it was able to deamidate acetamide, which differentiated it from all other species of Cupriavidus. DNA-DNA hybridization data revealed low levels of genomic DNA similarity (less than 30 %) between strain CPDB6(T) and the type strains of Cupriavidus species with validly published names. The major cellular fatty acids detected were cis-9-hexadecenoic (16 : 1omega7c) and hexadecanoic (16 : 0) acids. On the basis of phenotypic and genotypic characterizations, strain CPDB6(T) was recognized as a representative of a novel species within the genus Cupriavidus. The name Cupriavidus pampae sp. nov. is proposed, with strain CPDB6(T) (=CCUG 55948(T)=CCM-A-29:1289(T)) as the type strain. PMID- 20023054 TI - Sphingopyxis bauzanensis sp. nov., a psychrophilic bacterium isolated from soil. AB - Strain BZ30(T) was isolated from hydrocarbon-contaminated soil. The Gram negative, aerobic bacterium was psychrophilic and able to grow at temperatures ranging from 1 to 30  degrees C. The predominant cellular fatty acids of strain BZ30(T) were summed feature 3 (C(16 : 1)omega7c and/or iso-C(15 : 0) 2-OH) (37.4 %), C(18 : 1)omega7c (19.6 %), C(16 : 0) (8.2 %), C(14 : 0) 2-OH (8.0 %) and C(16 : 0) 2-OH (5.0 %). The predominant ubiquinone was Q-10. Major polar lipids were sphingoglycolipids, diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine and phosphatidylcholine. Spermidine was the major polyamine. The genomic DNA G+C content was 64.4 mol%. Phylogenetic analysis based on 16S rRNA gene sequence similarity showed that strain BZ30(T) belonged to the family Sphingomonadaceae of the alpha-4 group of the phylum Proteobacteria, and was related to the members of the genus Sphingopyxis, sharing the highest sequence similarities with the type strains of Sphingopyxis chilensis (98.3 %), S. witflariensis (98.2 %), S. taejonensis (97.4 %) and S. ginsengisoli (97.2 %). On the basis of the phenotypic characteristics, phylogenetic analysis and DNA-DNA relatedness data, strain BZ30(T) represents a novel species of the genus Sphingopyxis, for which the name Sphingopyxis bauzanensis is proposed. The type strain is BZ30(T) (=DSM 22271(T) =CGMCC 1.8959(T) =CIP 110136(T)). PMID- 20023055 TI - Description of Microvirga aerophila sp. nov. and Microvirga aerilata sp. nov., isolated from air, reclassification of Balneimonas flocculans Takeda et al. 2004 as Microvirga flocculans comb. nov. and emended description of the genus Microvirga. AB - Two bacterial strains, 5420S-12(T) and 5420S-16(T), isolated from air samples, were characterized using a polyphasic approach. 16S rRNA gene sequence analysis showed that strain 5420S-12(T) was related phylogenetically to Microvirga subterranea FaiI4(T) (97.4 % sequence similarity) and Microvirga guangxiensis 25B(T) (97.1 %) and that strain 5420S-16(T) was closely related to Balneimonas flocculans TFB(T) (98.0 %) and Microvirga guangxiensis 25B(T) (97.2 %). The G+C content of the genomic DNA was 62.2 mol% for strain 5420S-12(T) and 61.5 mol% for strain 5420S-16(T). The major fatty acid was C(18 : 1)omega7c. The results of DNA DNA hybridization and the phenotypic data showed that strains 5420S-12(T) and 5420S-16(T) could be distinguished from phylogenetically related species and represent two novel species within the genus Microvirga, for which the names Microvirga aerophila sp. nov. (type strain 5420S-12(T) =KACC 12743(T) =NBRC 106136(T)) and Microvirga aerilata sp. nov. (type strain 5420S-16(T) =KACC 12744(T) =NBRC 106137(T)) are proposed. Furthermore, the reclassification of Balneimonas flocculans as Microvirga flocculans comb. nov. (type strain TFB(T) =JCM 11936(T) =KCTC 12101(T) =IAM 15034(T) =ATCC BAA-817(T)) is proposed and an emended description of the genus Microvirga is provided. PMID- 20023056 TI - Photobacterium gaetbulicola sp. nov., a lipolytic bacterium isolated from a tidal flat sediment. AB - A Gram-negative, motile, non-spore-forming and lipolytic bacterial strain, designated Gung47(T), was isolated from a tidal flat on the west coast of Korea. Strain Gung47(T) grew optimally at 30 degrees C and with 2-5 % (w/v) NaCl. Phylogenetic analyses based on 16S rRNA gene sequences revealed that strain Gung47(T) belonged to the genus Photobacterium. Strain Gung47(T) exhibited 98.1 % 16S rRNA gene sequence similarity with Photobacterium rosenbergii LMG 22223(T) and 94.3-96.3 % similarity with other type strains of species of the genus Photobacterium. Strain Gung47(T) exhibited 47 % DNA-DNA relatedness to P. rosenbergii LMG 22223(T). Strain Gung47(T) contained Q-8 as the predominant ubiquinone and C(16 : 1)omega7c and/or iso-C(15 : 0) 2-OH, C(16 : 0) and C(18 : 1)omega7c as the major fatty acids. In this study, two closely related type strains, P. rosenbergii LMG 22223(T) and Photobacterium halotolerans LMG 22194(T), were also found to have Q-8 as the predominant ubiquinone. The DNA G+C content of strain Gung47(T) was 50.6 mol%. The differential phenotypic properties together with the phylogenetic and genetic distinctiveness of strain Gung47(T) demonstrated that this strain is distinguishable from recognized Photobacterium species. Therefore, strain Gung47(T) is considered to represent a novel species of the genus Photobacterium, for which the name Photobacterium gaetbulicola sp. nov. is proposed. The type strain is Gung47(T) (=KCTC 22804(T) =CCUG 58399(T)). PMID- 20023057 TI - Pedobacter bauzanensis sp. nov., isolated from soil. AB - A Gram-negative, aerobic, rod-shaped, non-motile bacterium, designated BZ42(T), was isolated from the soil of an industrial site. Strain BZ42(T) was able to grow at 5-25 degrees C. The major fatty acids were iso-C(15 : 0) (46.2 %), C(16 : 1)omega7c and/or iso-C(15 : 0) 2-OH (23.2 %) and iso-C(17 : 0) 3-OH (9.1 %). The predominant menaquinone was MK-7. The genomic DNA G+C content was 36.5 mol% (HPLC). 16S rRNA gene sequence phylogenetic analysis revealed that strain BZ42(T) was a member of the genus Pedobacter, family Sphingobacteriaceae, and 16S rRNA gene sequence similarities between strain BZ42(T) and the type strains of species of the genus Pedobacter with validly published names were 90.4-93.2 %. On the basis of phenotypic, chemotaxonomic and phylogenetic distinctiveness, strain BZ42(T) was considered to represent a novel species of the genus Pedobacter, for which the name Pedobacter bauzanensis sp. nov. is proposed. The type strain is BZ42(T) (=DSM 22554(T) =CGMCC 1.10187(T) =CIP 110134(T)). PMID- 20023058 TI - Kangiella japonica sp. nov., isolated from a marine environment. AB - Three Gram-negative, aerobic, halophilic, non-motile, yellowish-pigmented bacteria, designated KMM 3896, KMM 3897 and KMM 3899(T), were isolated from coastal seawater and sediment samples of the Sea of Japan, Russia. The isolates were shown to belong to the same species on the basis of 16S rRNA gene sequence similarities (99.6-99.9 %) and DNA-DNA relatedness (73-98 %). Phylogenetic analysis of 16S rRNA gene sequences demonstrated that the isolates formed a subcluster within the genus Kangiella in the class Gammaproteobacteria. 16S rRNA gene sequence similarities between strain KMM 3899(T) and its closest phylogenetic neighbours, Kangiella koreensis SW-125(T) and Kangiella aquimarina SW-154(T), were 96.6 and 95.5 %, respectively. On the basis of phenotypic differences and phylogenetic distances, it is proposed that strains KMM 3896, KMM 3897 and KMM 3899(T) are assigned to a novel species of the genus Kangiella, Kangiella japonica sp. nov. The type strain is KMM 3899(T) (=NRIC 0764(T) =JCM 16211(T)). PMID- 20023059 TI - Microbacterium lindanitolerans sp. nov., isolated from hexachlorocyclohexane contaminated soil. AB - A Gram-stain-positive, aerobic, rod-shaped, yellow actinobacterium, designated MNA2(T), was isolated from a hexachlorocyclohexane-contaminated soil in North India. Strain MNA2(T) showed 95 % 16S rRNA gene sequence similarity with Microbacterium halotolerans YIM 70130(T). Phylogenetic analysis of 16S rRNA gene sequences showed that strain MNA2(T) belonged to a clade represented by the genus Microbacterium of the family Microbacteriaceae. Strain MNA2(T) contained anteiso C(15 : 0), anteiso-C(17 : 0), iso-C(16 : 0) and iso-C(15 : 0) as the predominant fatty acids and diphosphatidylglycerol, phosphatidylglycerol and two unknown polar lipids. The menaquinones were MK-12, MK-11, MK-13 and MK-10, in an approximate molar ratio of 45 : 40 : 13 : 3, respectively. The DNA G+C content was 65.3 mol%. The peptidoglycan was of the B type of cross-linkage with ornithine as the diagnostic diamino acid. The results of the phylogenetic, phenotypic and chemotaxonomic analysis indicate that strain MNA2(T) belongs to a previously unrecognized species of the genus Microbacterium, for which the name Microbacterium lindanitolerans sp. nov. is proposed. The type strain is MNA2(T) (=DSM 22422(T) =CCM 7585(T)). PMID- 20023060 TI - Amycolatopsis helveola sp. nov. and Amycolatopsis pigmentata sp. nov., isolated from soil. AB - Three short spore chain-forming actinomycete strains were isolated from soil samples collected from subtropical islands in Japan. The cell-wall peptidoglycan of these strains contained meso-diaminopimelic acid (meso-A(2)pm), glutamic acid and alanine. The major isoprenoid quinone was MK-9(H(4)), iso-C(16 : 0) and 2-OH iso-C(16 : 0) were the major cellular fatty acids and phosphatidylethanolamine was a component of the polar lipids. The G+C content of the genomic DNA was 67-69 mol%. Phylogenetic analyses based on the 16S rRNA gene sequences showed that the novel strains consistently formed a monophyletic cluster with Amycolatopsis taiwanensis. On the basis this polyphasic taxonomical study, it is proposed that the two new isolates represent two novel species: Amycolatopsis helveola (type strain TT00-43(T)=NBRC 103394(T)=KCTC 19329(T)) and Amycolatopsis pigmentata (type strain TT99-32(T)=NBRC 103392(T)=KCTC 19330(T)). PMID- 20023061 TI - Slackia piriformis sp. nov. and Collinsella tanakaei sp. nov., new members of the family Coriobacteriaceae, isolated from human faeces. AB - Three Gram-positive, strictly anaerobic, non-spore-forming, rod-shaped organisms (strains YIT 12062(T), YIT 12063(T) and YIT 12064) were isolated from human faeces. Strain YIT 12062(T) was asaccharolytic and possessed a DNA G+C content of 58.3 mol%. Cells of strain YIT 12062(T) were negative for catalase, oxidase, urease, hydrolysis of aesculin and gelatin, nitrate reduction and indole production. Based on 16S rRNA gene sequence analysis, strain YIT 12062(T) was assigned to the genus Slackia (91.7-96.0 % sequence similarities to type strains of Slackia species). Biochemical data showed that the isolate was phenotypically distinct from all recognized species of the genus Slackia. Strain YIT 12062(T) therefore represents a novel species in the genus Slackia, for which the name Slackia piriformis sp. nov. is proposed. The type strain is YIT 12062(T) (=DSM 22477(T)=JCM 16070(T)). Following 16S rRNA gene sequence analysis, strains YIT 12063(T) and YIT 12064, which were isolated from different subjects, were shown to be most closely related to species of the genus Collinsella (93.8-95.1 % similarities to type strains). Although their phenotypic characteristics were very similar and they shared >99 % 16S rRNA gene sequence similarity and >97+/ 1.8 % DNA-DNA relatedness, the two isolates could be discriminated by RAPD fingerprints. The DNA G+C contents of strains YIT 12063(T) and YIT 12064 were 60.8 and 61.0 mol%, respectively. They were saccharolytic in API test systems, positive for aesculin hydrolysis and negative for catalase, oxidase, urease, indole production, nitrate reduction and gelatin hydrolysis. The major end products of glucose fermentation of these strains were lactate, acetate and formate. Biochemical data supported the affiliation of strains YIT 12063(T) and YIT 12064 to the genus Collinsella and showed that they were phenotypically distinct from all recognized species of the genus Collinsella. Strains YIT 12063(T) and YIT 12064 therefore represent a novel species of the genus Collinsella, for which the name Collinsella tanakaei sp. nov. is proposed. The type strain is YIT 12063(T) (=DSM 22478(T)=JCM 16071(T)). PMID- 20023062 TI - Methylophilus flavus sp. nov. and Methylophilus luteus sp. nov., aerobic, methylotrophic bacteria associated with plants. AB - Novel yellow, obligately methylotrophic and restricted facultatively methylotrophic bacteria, respectively designated strains Ship(T) and Mim(T), with the ribulose monophosphate pathway of C(1) assimilation are described. Cells were strictly aerobic, Gram-negative, asporogenous, non-motile rods that multiply by binary fission, were mesophilic and neutrophilic and synthesized indole-3-acetic acid and exopolysaccharide. The predominant cellular fatty acids were C(16 : 0) and C(16 : 1). The major ubiquinone was Q-8. The predominant phospholipids were phosphatidylethanolamine and phosphatidylglycerol; diphosphatidylglycerol was absent. The two strains lacked alpha-ketoglutarate dehydrogenase and glutamate dehydrogenase. They assimilated ammonium via the glutamate cycle enzymes glutamine synthetase and glutamate synthase. The DNA G+C contents of strains Ship(T) and Mim(T) were 50.7 and 54.5 mol% (T(m)), respectively. The level of 16S rRNA gene sequence similarity between these strains was very high (99.8 %) but they shared a low level of DNA-DNA relatedness (44 %). Based on 16S rRNA gene sequence analysis and low levels of DNA-DNA relatedness with the type strains of recognized species of the genus Methylophilus (31-36 %), strains Ship(T) and Mim(T) are considered to represent novel species of the genus Methylophilus, for which the names Methylophilus flavus sp. nov. (type strain Ship(T) =DSM 23073(T) =VKM B-2547(T) =CCUG 58411(T)) and Methylophilus luteus sp. nov. (type strain Mim(T) =DSM 22949(T) =VKM B-2548(T) =CCUG 58412(T)) are proposed. PMID- 20023063 TI - Halogeometricum rufum sp. nov., a halophilic archaeon from a marine solar saltern, and emended description of the genus Halogeometricum. AB - Two halophilic archaea, strains RO1-4(T) and RO1-64, were isolated from a marine solar saltern in Jiangsu, China. Cells of the two strains were pleomorphic, motile, and stained Gram-negative. Colonies were red-pigmented. Strains RO1-4(T) and RO1-64 were able to grow at 25-55 degrees C (optimum 40-42 degrees C), at 2.1-5.1 M NaCl (optimum 3.9 M NaCl), at 0.05-0.7 M MgCl(2) (optimum 0.3 M MgCl(2)) and at pH 6.0-8.5 (optimum pH 7.0). Cells lyse in distilled water and the minimal NaCl concentration to prevent cell lysis is 12 % (w/v). On the basis of 16S rRNA gene sequence analysis, strains RO1-4(T) and RO1-64 were closely related to Halogeometricum borinquense PR3(T) (98.0 and 98.2 % similarity, respectively) and Halosarcina pallida BZ256(T) (97.8 and 97.9 %). The major polar lipids of the two strains were phosphatidylglycerol, phosphatidylglycerol phosphate methyl ester and two glycolipids (S-DGD-1 and DGD-1). The DNA G+C contents of strains RO1-4(T) and RO1-64 are 64.9 and 62.4 mol%, respectively. The DNA-DNA hybridization value between strains RO1-4(T) and RO1-64 was 83.0 % and both strains showed low DNA-DNA relatedness with Halogeometricum borinquense PR3(T) (42.5 and 50.1 % relatedness, respectively) and Halosarcina pallida BZ256(T) (37.6 and 42.1 % relatedness). It was concluded that strains RO1-4(T) and RO1-64 represent a novel species of the genus Halogeometricum, for which the name Halogeometricum rufum sp. nov. is proposed. The type strain is RO1-4(T) (=CGMCC 1.7736(T) =JCM 15770(T)). PMID- 20023064 TI - Orbus hercynius gen. nov., sp. nov., isolated from faeces of wild boar, is most closely related to members of the orders 'Enterobacteriales' and Pasteurellales. AB - A novel gammaproteobacterium, strain CN3(T), was isolated from the faeces of wild boar. Strain CN3(T) was facultatively anaerobic and appeared coccoid or rod shaped. The partial 16S rRNA gene sequence determined for strain CN3(T) suggested a distant relationship with members of the orders 'Enterobacteriales' and Pasteurellales. The gene sequence showed highest similarity (90.3 %) with Obesumbacterium proteus DSM 2777(T), a member of the family Enterobacteriaceae. The closest relatives outside the order 'Enterobacteriales' according to 16S rRNA gene sequence analysis were members of the order Pasteurellales with 88.7 % similarity (Mannheimia haemolytica NCTC 9380(T) and Actinobacillus lignieresii NCTC 4189(T)). In contrast to most members of the order 'Enterobacteriales', strain CN3(T) was oxidase-positive. The pattern of fatty acids, in particular the high relative abundance of C(18 : 1)omega7c (38.5 %), was clearly distinct from the conserved pattern found for members of the order Pasteurellales. EcoRI ribotyping of strain CN3(T) yielded no significant similarity to existing database entries. The major ubiquinone of strain CN3(T) was Q-8. The DNA G+C content was 36.4 mol%. Strain CN3(T) hosted a phage and secreted considerable amounts of three proteins into the culture supernatant. A spontaneous mutant of strain CN3(T) was isolated which formed long filaments. Microscopic studies revealed the presence of a capsule that the mutant strain was unable to partition after cell division. Strain CN3(T) thus represents a novel species within a new genus, for which the name Orbus hercynius gen. nov., sp. nov. is proposed. The type strain of the type species is CN3(T) (=DSM 22228(T)=CCUG 57622(T)). Classification of the novel species to the family and order level will require further investigations. PMID- 20023065 TI - Increased risk of death among children with Lennox-Gastaut syndrome and infantile spasms. AB - The magnitude and causes of death among a cohort of children with epilepsy were determined. A follow-up study with a population-based cohort of 10-year-old children in the metropolitan Atlanta area with epilepsy was conducted. The National Death Index and linkage to State of Georgia death certificates were used to identify deaths. The authors estimated the expected numbers of deaths by applying mortality rates adjusted by age, race, and sex for the entire state of Georgia to the population for the follow-up period. Among the 688 children who were in the final epilepsy cohort, 64 deaths occurred; 20.6 deaths were expected (mortality ratio adjusted for age, race, and sex = 3.11). The mortality ratios for children with Lennox-Gastaut syndrome and infantile spasms were 13.92 and 11.91, respectively. Children and adolescents with epilepsy, especially those with Lennox-Gastaut syndrome or infantile spasms, have an increased risk of death. PMID- 20023066 TI - Diagnosis, treatment, and long-term outcomes of late-onset (type III) multiple acyl-CoA dehydrogenase deficiency. AB - We report 4 children with late-onset (type III) multiple acyl-CoA dehydrogenase deficiency, also known as glutaric aciduria type II, which is an autosomal recessive disorder of fatty acid and amino acid metabolism. The underlying deficiency is in the electron transfer flavoprotein or electron flavoprotein dehydrogenase. Clinical presentations include fatal acute neonatal metabolic encephalopathies with/without organ system anomalies (types I and II) and late onset acute metabolic crises, myopathy, or neurodevelopmental delays (type III). Two patients were identified in childhood following a metabolic crisis and/or neurodevelopmental delay, and 2 were identified by newborn metabolic screening. Our cases will illustrate the difficulty in making a biochemical diagnosis of late-onset (type III) multiple acyl-CoA dehydrogenase deficiency from plasma acylcarnitines and urine organic acids in both symptomatic and asymptomatic children. However, they emphasize the need for timely diagnosis to urgently implement prophylactic treatment for life-threatening metabolic crises with low protein/fat diets supplemented with riboflavin and carnitine. PMID- 20023067 TI - Comparative transcript profiling of Candida albicans and Candida dubliniensis identifies SFL2, a C. albicans gene required for virulence in a reconstituted epithelial infection model. AB - Candida albicans and Candida dubliniensis are closely related species displaying differences in virulence and genome content, therefore providing potential opportunities to identify novel C. albicans virulence genes. C. albicans gene arrays were used for comparative analysis of global gene expression in the two species in reconstituted human oral epithelium (RHE). C. albicans (SC5314) showed upregulation of hypha-specific and virulence genes within 30 min postinoculation, coinciding with rapid induction of filamentation and increased RHE damage. C. dubliniensis (CD36) showed no detectable upregulation of hypha-specific genes, grew as yeast, and caused limited RHE damage. Several genes absent or highly divergent in C. dubliniensis were upregulated in C. albicans. One such gene, SFL2 (orf19.3969), encoding a putative heat shock factor, was deleted in C. albicans. DeltaDeltasfl2 cells failed to filament under a range of hypha-inducing conditions and exhibited greatly reduced RHE damage, reversed by reintroduction of SFL2 into the DeltaDeltasfl2 strain. Moreover, SFL2 overexpression in C. albicans triggered hyphal morphogenesis. Although SFL2 deletion had no apparent effect on host survival in the murine model of systemic infection, DeltaDeltasfl2 strain-infected kidney tissues contained only yeast cells. These results suggest a role for SFL2 in morphogenesis and an indirect role in C. albicans pathogenesis in epithelial tissues. PMID- 20023068 TI - Candida albicans PEP12 is required for biofilm integrity and in vivo virulence. AB - To investigate the role of the prevacuolar secretion pathway in biofilm formation and virulence in Candida albicans, we cloned and analyzed the C. albicans homolog of the Saccharomyces cerevisiae prevacuolar trafficking gene PEP12. C. albicans PEP12 encodes a deduced t-SNARE that is 28% identical to S. cerevisiae Pep12p, and plasmids bearing C. albicans PEP12 complemented the abnormal vacuolar morphology and temperature-sensitive growth of an S. cerevisiae pep12 null mutant. The C. albicans pep12 Delta null mutant was defective in endocytosis and vacuolar acidification and accumulated 40- to 60-nm cytoplasmic vesicles near the plasma membrane. Secretory defects included increased extracellular proteolytic activity and absent lipolytic activity. The pep12Delta null mutant was more sensitive to cell wall stresses and antifungal agents than the isogenic complemented strain or the control strain DAY185. Notably, the biofilm formed by the pep12Delta mutant was reduced in overall mass and fragmented completely upon the slightest disturbance. The pep12Delta mutant was markedly reduced in virulence in an in vitro macrophage infection model and an in vivo mouse model of disseminated candidiasis. These results suggest that C. albicans PEP12 plays a key role in biofilm integrity and in vivo virulence. PMID- 20023069 TI - Hgc1 mediates dynamic Candida albicans-endothelium adhesion events during circulation. AB - Common iatrogenic procedures can result in translocation of the human pathogenic fungus Candida albicans from mucosal surfaces to the bloodstream. Subsequent disseminated candidiasis and infection of deep-seated organs may occur if the fungus is not eliminated by blood cells. In these cases, fungal cells adhere to the endothelial cells of blood vessels, penetrate through endothelial layers, and invade deeper tissue. In this scenario, endothelial adhesion events must occur during circulation under conditions of physiological blood pressure. To investigate the fungal and host factors which contribute to this essential step of disseminated candidiasis, we have developed an in vitro circulatory C. albicans-endothelium interaction model. We demonstrate that both C. albicans yeast and hyphae can adhere under flow at a pressure similar to capillary blood pressure. Serum factors significantly enhanced the adhesion potential of viable but not killed C. albicans cells to endothelial cells. During circulation, C. albicans cells produced hyphae and the adhesion potential first increased, then decreased with time. We provide evidence that a specific temporal event in the yeast-to-hyphal transition, regulated by the G(1) cyclin Hgc1, is critical for C. albicans-endothelium adhesion during circulation. PMID- 20023070 TI - Distinct subcellular localization of a group of synaptobrevin-like SNAREs in Paramecium tetraurelia and effects of silencing SNARE-specific chaperone NSF. AB - We have identified new synaptobrevin-like SNAREs and localized the corresponding gene products with green fluorescent protein (GFP)-fusion constructs and specific antibodies at the light and electron microscope (EM) levels. These SNAREs, named Paramecium tetraurelia synaptobrevins 8 to 12 (PtSyb8 to PtSyb12), showed mostly very restricted, specific localization, as they were found predominantly on structures involved in endo- or phagocytosis. In summary, we found PtSyb8 and PtSyb9 associated with the nascent food vacuole, PtSyb10 near the cell surface, at the cytostome, and in close association with ciliary basal bodies, and PtSyb11 on early endosomes and on one side of the cytostome, while PtSyb12 was found in the cytosol. PtSyb4 and PtSyb5 (identified previously) were localized on small vesicles, PtSyb5 probably being engaged in trichocyst (dense core secretory vesicle) processing. PtSyb4 and PtSyb5 are related to each other and are the furthest deviating of all SNAREs identified so far. Because they show no similarity with any other R-SNAREs outside ciliates, they may represent a ciliate specific adaptation. PtSyb10 forms small domains near ciliary bases, and silencing slows down cell rotation during depolarization-induced ciliary reversal. NSF silencing supports a function of cell surface SNAREs by revealing vesicles along the cell membrane at sites normally devoid of vesicles. The distinct distributions of these SNAREs emphasize the considerable differentiation of membrane trafficking, particularly along the endo-/phagocytic pathway, in this protozoan. PMID- 20023071 TI - Phospholipid-binding protein EhC2A mediates calcium-dependent translocation of transcription factor URE3-BP to the plasma membrane of Entamoeba histolytica. AB - The Entamoeba histolytica upstream regulatory element 3-binding protein (URE3-BP) is a transcription factor that binds DNA in a Ca(2+)-inhibitable manner. The protein is located in both the nucleus and the cytoplasm but has also been found to be enriched in the plasma membrane of amebic trophozoites. We investigated the reason for the unusual localization of URE3-BP at the amebic plasma membrane. Here we identify and characterize a 22-kDa Ca(2+)-dependent binding partner of URE3-BP, EhC2A, a novel member of the C2-domain superfamily. Immunoprecipitations of URE3-BP and EhC2A showed that the proteins interact and that such interaction was enhanced in the presence of Ca(2+). Recombinant and native EhC2A bound phospholipid liposomes in a Ca(2+)-dependent manner, with half-maximal binding occurring at 3.4 muM free Ca(2+). A direct interaction between EhC2A and URE3-BP was demonstrated by the ability of recombinant EhC2A to recruit recombinant URE3 BP to phospholipid liposomes in a Ca(2+)-dependent manner. URE3-BP and EhC2A were observed to translocate to the amebic plasma membrane upon an increase in the intracellular Ca(2+) concentration of trophozoites, as revealed by subcellular fractionation and immunofluorescent staining. Short hairpin RNA-mediated knockdown of EhC2A protein expression significantly modulated the mRNA levels of URE3-BP-regulated transcripts. Based on these results, we propose a model for EhC2A-mediated regulation of the transcriptional activities of URE3-BP via Ca(2+) dependent anchoring of the transcription factor to the amebic plasma membrane. PMID- 20023072 TI - tRNA gene sequences are required for transcriptional silencing in Entamoeba histolytica. AB - Transcriptional silencing by trans inactivation can contribute to the regulation of gene expression in eukaryotic cells. In the human intestinal protozoan parasite Entamoeba histolytica, trans inactivation of the amoebapore-A gene (AP A) was recently achieved by episomal transfection of E. histolytica trophozoites with the plasmid psAP1. The mechanism of AP-A trans inactivation is largely unknown, though it was suggested that a partial short interspersed transposable element (SINE) is required. By systematic assessment of various E. histolytica isolates transfected with psAP1 derivates, trans inactivation of AP-A was restricted to the strain HM-1:IMSS (2411) but could not be achieved in other standard laboratory strains. Importantly, sequences of an E. histolytica tRNA array that were located on psAP1 in close proximity to the AP-A upstream region and comprising the glutamic acid (TTC) (E) and tyrosine (GTA) (Y) tRNA genes were indispensable for AP-A silencing. In contrast to the case described in previous reports, SINE was not required for AP-A trans inactivation. AP-A expression could be regained in silenced cells by episomal transfection under the control of a heterologous E. histolytica promoter, opening a way toward future silencing of individual genes of interest in E. histolytica. Our results indicate that tRNA gene-mediated silencing is not restricted to Saccharomyces cerevisiae. PMID- 20023074 TI - Studying the dynamics of flagella in multicellular communities of Escherichia coli by using biarsenical dyes. AB - This paper describes a new approach for labeling intact flagella using the biarsenical dyes FlAsH and ReAsH and imaging their spatial and temporal dynamics on live Escherichia coli cells in swarming communities of bacteria by using epifluorescence microscopy. Using this approach, we observed that (i) bundles of flagella on swarmer cells remain cohesive during frequent collisions with neighboring cells, (ii) flagella on nonmotile swarmer cells at the leading edge of the colony protrude in the direction of the uncolonized agar surface and are actively rotated in a thin layer of fluid that extends outward from the colony, and (iii) flagella form transient interactions with the flagella of other swarmer cells that are in close proximity. This approach opens a window for observing the dynamics of cells in communities that are relevant to ecology, industry, and biomedicine. PMID- 20023075 TI - Milk protein fragments induce the biosynthesis of macedocin, the lantibiotic produced by Streptococcus macedonicus ACA-DC 198. AB - The aim of the present work was to study the mode of the induction of the biosynthesis of macedocin, the lantibiotic produced by Streptococcus macedonicus ACA-DC 198. Macedocin was produced when the strain was grown in milk but not in MRS or M17 broth. No autoinduction mechanism was observed. Production did not depend on the presence of lactose or galactose in the culture medium or on a coculture of the producer strain with macedocin-sensitive or macedocin-resistant strains. Induction seemed to depend on the presence of one or more heat-stable protein components produced when S. macedonicus ACA-DC 198 was grown in milk. The partial purification of the induction factor was performed by a combination of chromatography methods, and its activity was confirmed by a reverse transcription PCR approach (RT-PCR). Mass spectrometric (MS) and tandem mass spectrometric (MS/MS) analyses of an induction-active fraction showed the presence of several peptides of low molecular mass corresponding to fragments of alpha(S1)- and beta casein as well as beta-lactoglobulin. The chemically synthesized alpha(S1)-casein fragment 37-55 (2,253.65 Da) was proven to be able to induce macedocin biosynthesis. This is the first time that milk protein degradation fragments are reported to exhibit a bacteriocin induction activity. PMID- 20023076 TI - Correlation between environmental factors and prevalence of Vibrio parahaemolyticus in oysters harvested in the southern coastal area of Sao Paulo State, Brazil. AB - The presence of Vibrio parahaemolyticus in 123 oyster samples collected from an estuary on the southern coast of Sao Paulo state, Brazil, was investigated. Of the 123 samples, 99.2% were positive with densities ranging from < 3 to 10(5) most probable number (MPN)/g. Densities correlated significantly with water temperature (r = 0.48; P < 0.001) but not with salinity (r = -0.09; P = 0.34). The effect of harvest site on counts was not significant (P > 0.05). These data provide information for the assessment of exposure of V. parahaemolyticus in oysters at harvest. PMID- 20023077 TI - Association of novel and highly diverse acid-tolerant denitrifiers with N2O fluxes of an acidic fen. AB - Wetlands are sources of denitrification-derived nitrous oxide (N2O). Thus, the denitrifier community of an N2O-emitting fen (pH 4.7 to 5.2) was investigated. N2O was produced and consumed to subatmospheric concentrations in unsupplemented anoxic soil microcosms. Total cell counts and most probable numbers of denitrifiers approximated 10(11) cells x g(DW)(-1) (where DW is dry weight) and 10(8) cells x g(DW)(-1), respectively, in both 0- to 10-cm and 30- to 40-cm depths. Despite this uniformity, depth-related maximum reaction rate (v(max)) values for denitrification in anoxic microcosms ranged from 1 to 24 and -19 to 105 nmol N2O h(-1) x g(DW)(-1), with maximal values occurring in the upper soil layers. Denitrification was enhanced by substrates that might be formed via fermentation in anoxic microzones of soil. N2O approximated 40% of total nitrogenous gases produced at in situ pH, which was likewise the optimal pH for denitrification. Gene libraries of narG and nosZ (encoding nitrate reductase and nitrous oxide reductase, respectively) from fen soil DNA yielded 15 and 18 species-level operational taxonomic units, respectively, many of which displayed phylogenetic novelty and were not closely related to cultured organisms. Although statistical analyses of narG and nosZ sequences indicated that the upper 20 cm of soil contained the highest denitrifier diversity and species richness, terminal restriction fragment length polymorphism analyses of narG and nosZ revealed only minor differences in denitrifier community composition from a soil depth of 0 to 40 cm. The collective data indicate that the regional fen harbors novel, highly diverse, acid-tolerant denitrifier communities capable of complete denitrification and consumption of atmospheric N2O at in situ pH. PMID- 20023078 TI - Metabolic differentiation in biofilms as indicated by carbon dioxide production rates. AB - The measurement of carbon dioxide production rates as an indication of metabolic activity was applied to study biofilm development and response of Pseudomonas sp. biofilms to an environmental disturbance in the form of a moving air-liquid interface (i.e., shear). A differential response in biofilm cohesiveness was observed after bubble perturbation, and the biofilm layers were operationally defined as either shear-susceptible or non-shear-susceptible. Confocal laser scanning microscopy and image analysis showed a significant reduction in biofilm thickness and biomass after the removal of the shear-susceptible biofilm layer, as well as notable changes in the roughness coefficient and surface-to-biovolume ratio. These changes were accompanied by a 72% reduction of whole-biofilm CO2 production; however, the non-shear-susceptible region of the biofilm responded rapidly after the removal of the overlying cells and extracellular polymeric substances (EPS) along with the associated changes in nutrient and O2 flux, with CO2 production rates returning to preperturbation levels within 24 h. The adaptable nature and the ability of bacteria to respond to environmental conditions were further demonstrated by the outer shear-susceptible region of the biofilm; the average CO2 production rate of cells from this region increased within 0.25 h from 9.45 +/- 5.40 fmol of CO2 x cell(-1) x h(-1) to 22.6 +/- 7.58 fmol of CO2 x cell(-1) x h(-1) when cells were removed from the biofilm and maintained in suspension without an additional nutrient supply. These results also demonstrate the need for sufficient monitoring of biofilm recovery at the solid substratum if mechanical methods are used for biofouling control. PMID- 20023080 TI - Inactivation of adenoviruses, enteroviruses, and murine norovirus in water by free chlorine and monochloramine. AB - Inactivation of infectious viruses during drinking water treatment is usually achieved with free chlorine. Many drinking water utilities in the United States now use monochloramine as a secondary disinfectant to minimize disinfectant by product formation and biofilm growth. The inactivation of human adenoviruses 2, 40, and 41 (HAdV2, HAdV40, and HAdV41), coxsackieviruses B3 and B5 (CVB3 and CVB5), echoviruses 1 and 11 (E1 and E11), and murine norovirus (MNV) are compared in this study. Experiments were performed with 0.2 mg of free chlorine or 1 mg of monochloramine/liter at pH 7 and 8 in buffered reagent-grade water at 5 degrees C. CT values (disinfectant concentration x time) for 2- to 4-log(10) (99 to 99.99%) reductions in virus titers were calculated by using the efficiency factor Hom model. The enteroviruses required the longest times for chlorine inactivation and MNV the least time. CVB5 required the longest exposure time, with CT values of 7.4 and 10 mg x min/liter (pH 7 and 8) for 4-log(10) inactivation. Monochloramine disinfection was most effective for E1 (CT values ranged from 8 to 18 mg x min/liter for 2- and 3-log(10) reductions, respectively). E11 and HAdV2 were the least susceptible to monochloramine disinfection (CT values of 1,300 and 1,600 mg-min/liter for 3-log(10) reductions, respectively). Monochloramine inactivation was most successful for the adenoviruses, CVB5, and E1 at pH 7. A greater variation in inactivation rates between viruses was observed during monochloramine disinfection than during chlorine disinfection. These data will be useful in drinking water risk assessment studies and disinfection system planning. PMID- 20023079 TI - Archaeal diversity and distribution along thermal and geochemical gradients in hydrothermal sediments at the Yonaguni Knoll IV hydrothermal field in the Southern Okinawa trough. AB - A variety of archaeal lineages have been identified using culture-independent molecular phylogenetic surveys of microbial habitats occurring in deep-sea hydrothermal environments such as chimney structures, sediments, vent emissions, and chemosynthetic macrofauna. With the exception of a few taxa, most of these archaea have not yet been cultivated, and their physiological and metabolic traits remain unclear. In this study, phylogenetic diversity and distribution profiles of the archaeal genes encoding small subunit (SSU) rRNA, methyl coenzyme A (CoA) reductase subunit A, and the ammonia monooxygenase large subunit were characterized in hydrothermally influenced sediments at the Yonaguni Knoll IV hydrothermal field in the Southern Okinawa Trough. Sediment cores were collected at distances of 0.5, 2, or 5 m from a vent emission (90 degrees C). A moderate temperature gradient extends both horizontally and vertically (5 to 69 degrees C), indicating the existence of moderate mixing between the hydrothermal fluid and the ambient sediment pore water. The mixing of reductive hot hydrothermal fluid and cold ambient sediment pore water establishes a wide spectrum of physical and chemical conditions in the microbial habitats that were investigated. Under these different physico-chemical conditions, variability in archaeal phylotype composition was observed. The relationship between the physical and chemical parameters and the archaeal phylotype composition provides important insight into the ecophysiological requirements of uncultivated archaeal lineages in deep-sea hydrothermal vent environments, giving clues for approximating culture conditions to be used in future culturing efforts. PMID- 20023081 TI - A mariner-based transposon system for in vivo random mutagenesis of Clostridium difficile. AB - Understanding the molecular basis of Clostridium difficile infection is a prerequisite to the development of effective countermeasures. Although there are methods for constructing gene-specific mutants of C. difficile, currently there is no effective method for generating libraries of random mutants. In this study, we developed a novel mariner-based transposon system for in vivo random mutagenesis of C. difficile R20291, the BI/NAP1/027 epidemic strain at the center of the C. difficile outbreaks in Stoke Mandeville, United Kingdom, in 2003 to 2004 and 2004 to 2005. Transposition occurred at a frequency of 4.5 (+/-0.4) x 10(-4) per cell to give stable insertions at random genomic loci, which were defined only by the nucleotide sequence TA. Furthermore, mutants with just a single transposon insertion were generated in an overwhelming majority (98.3% in this study). Phenotypic screening of a C. difficile R20291 random mutant library yielded a sporulation/germination-defective clone with an insertion in the germination-specific protease gene cspBA and an auxotroph with an insertion in the pyrimidine biosynthesis gene pyrB. These results validate our mariner-based transposon system for use in forward genetic studies of C. difficile. PMID- 20023082 TI - Type II diacylglycerol acyltransferase from Claviceps purpurea with ricinoleic acid, a hydroxyl fatty acid of industrial importance, as preferred substrate. AB - Claviceps purpurea, the fungal pathogen that causes the cereal disease ergot, produces glycerides that contain high levels of ricinoleic acid [(R)-12 hydroxyoctadec-cis-9-enoic acid] in its sclerotia. Recently, a fatty acid hydroxylase (C. purpurea FAH [CpFAH]) involved in the biosynthesis of ricinoleic acid was identified from this fungus (D. Meesapyodsuk and X. Qiu, Plant Physiol. 147:1325-1333, 2008). Here, we describe the cloning and biochemical characterization of a C. purpurea type II diacylglycerol acyltransferase (CpDGAT2) involved in the assembly of ricinoleic acid into triglycerides. The CpDGAT2 gene was cloned by degenerate RT-PCR (reverse transcription-PCR). The expression of this gene restored the in vivo synthesis of triacylglycerol (TAG) in the quadruple mutant strain Saccharomyces cerevisiae H1246, in which all four TAG biosynthesis genes (DGA1, LRO1, ARE1, and ARE2) are disrupted. In vitro enzymatic assays using microsomal preparations from the transformed yeast strain indicated that CpDGAT2 prefers ricinoleic acid as an acyl donor over linoleic acid, oleic acid, or linolenic acid, and it prefers 1,2-dioleoyl-sn-glycerol over 1,2-dipalmitoyl-sn-glycerol as an acyl acceptor. The coexpression of CpFAH with CpDGAT2 in yeast resulted in an increased accumulation of ricinoleic acid compared to the coexpression of CpFAH with the native yeast DGAT2 (S. cerevisiae DGA1 [ScDGA1]) or the expression of CpFAH alone. Northern blot analysis indicated that CpFAH is expressed solely in sclerotium cells, with no transcripts of this gene being detected in mycelium or conidial cells. CpDGAT2 was more widely expressed among the cell types examined, although expression was low in conidiospores. The high expression of CpDGAT2 and CpFAH in sclerotium cells, where high levels of ricinoleate glycerides accumulate, provided further evidence supporting the roles of CpDGAT2 and CpFAH as key enzymes for the synthesis and assembly of ricinoleic acid in C. purpurea. PMID- 20023083 TI - Indirect effects of temperature on stink bug fitness, via maintenance of gut associated symbionts. AB - Impacts of climate change on organisms are already apparent, with effects ranging from the individual to ecosystem scales. For organisms engaged in mutualisms, climate may affect population performance directly or indirectly through mediated effects on their mutualists. We tested this hypothesis for two stink bugs, Acrosternum hilare and Murgantia histrionica, and their gut-associated symbionts. We reared these species at two constant temperatures, 25 and 30 degrees C, and monitored population demographic parameters and the presence of gut-associated symbionts with diagnostic PCR primer sets. Both stink bugs lost their respective gut symbionts within two generations at 30 degrees C. In addition, the insect survivorship and reproductive rates of both A. hilare and M. histrionica at 30 degrees C were lower than at 25 degrees C. Other demographic parameters also indicated a decrease in overall insect fitness at the high temperature. Collectively our data showed that the decrease in host fitness was coupled with, and potentially mediated by, symbiont loss at 30 degrees C. This work illustrates the need to better understand the biology of animal-symbiont associations and the consequences of local climate for the dynamics of these interactions. PMID- 20023084 TI - Improved electrotransformation and decreased antibiotic resistance of the cystic fibrosis pathogen Burkholderia cenocepacia strain J2315. AB - The bacterium Burkholderia cenocepacia is pathogenic for sufferers from cystic fibrosis (CF) and certain immunocompromised conditions. The B. cenocepacia strain most frequently isolated from CF patients, and which serves as the reference for CF epidemiology, is J2315. The J2315 genome is split into three chromosomes and one plasmid. The strain was sequenced several years ago, and its annotation has been released recently. This information should allow genetic experimentation with J2315, but two major impediments appear: the poor potential of J2315 to act as a recipient in transformation and conjugation and the high level of resistance it mounts to nearly all antibiotics. Here, we describe modifications to the standard electroporation procedure that allow routine transformation of J2315 by DNA. In addition, we show that deletion of an efflux pump gene and addition of spermine to the medium enhance the sensitivity of J2315 to certain commonly used antibiotics and so allow a wider range of antibiotic resistance genes to be used for selection. PMID- 20023085 TI - Shear-enhanced oral microbial adhesion. AB - Shear-enhanced adhesion, although not observed for fimbria-mediated adhesion of oral Actinomyces spp., was noted for Hsa-mediated adhesion of Streptococcus gordonii to sialic acid-containing receptors, an interaction implicated in the pathogenesis of infective endocarditis. PMID- 20023086 TI - Optimization of RNA extraction for PCR quantification of aromatic compound degradation genes. AB - Seven different bacterial strains and primer sets and a mixed community were used to evaluate the use of reverse transcriptase quantitative PCR (Q-PCR) and Q-PCR of oxygenase genes to assess various approaches for monitoring the bioremediation of polluted sites. Differences in maximum activity were seen when different RNA extraction kits were compared. PMID- 20023087 TI - Transfer of a phage T4 gene into Enterobacteriaceae, determined at the single cell level. AB - The transfer range of phage genes was investigated at the single-cell level by using an in situ DNA amplification technique. After absorption of phages, a phage T4 gene was maintained in the genomes of non-plaque-forming bacteria at frequencies of 10(-2) gene copies per cell. The gene transfer decreased the mutation frequencies in nonhost recipients. PMID- 20023088 TI - Thermococcus kodakarensis genetics: TK1827-encoded beta-glycosidase, new positive selection protocol, and targeted and repetitive deletion technology. AB - Inactivation of TK1761, the reporter gene established for Thermococcus kodakarensis, revealed the presence of a second beta-glycosidase that we have identified as the product of TK1827. This enzyme (pTK1827) has been purified and shown to hydrolyze glucopyranoside but not mannopyranoside, have optimal activity at 95 degrees C and from pH 8 to 9.5, and have a functional half-life of approximately 7 min at 100 degrees C. To generate a strain with both TK1761 and TK1827 deleted, a new selection/counterselection protocol has been developed, and the levels of beta-glycosidase activity in T. kodakarensis strains with TK1761 and/or TK1827 deleted and with these genes expressed from heterologous promoters are described. Genetic tools and strains have been developed that extend the use of this selection/counterselection procedure to delete any nonessential gene from the T. kodakarensis chromosome. Using this technology, TK0149 was deleted to obtain an agmatine auxotroph that grows on nutrient-rich medium only when agmatine is added. Transformants can therefore be selected rapidly, and replicating plasmids can be maintained in this strain growing in rich medium by complementation of the TK0149 deletion. PMID- 20023089 TI - Soil microbial community responses to multiple experimental climate change drivers. AB - Researchers agree that climate change factors such as rising atmospheric [CO2] and warming will likely interact to modify ecosystem properties and processes. However, the response of the microbial communities that regulate ecosystem processes is less predictable. We measured the direct and interactive effects of climatic change on soil fungal and bacterial communities (abundance and composition) in a multifactor climate change experiment that exposed a constructed old-field ecosystem to different atmospheric CO2 concentration (ambient, +300 ppm), temperature (ambient, +3 degrees C), and precipitation (wet and dry) might interact to alter soil bacterial and fungal abundance and community structure in an old-field ecosystem. We found that (i) fungal abundance increased in warmed treatments; (ii) bacterial abundance increased in warmed plots with elevated atmospheric [CO2] but decreased in warmed plots under ambient atmospheric [CO2]; (iii) the phylogenetic distribution of bacterial and fungal clones and their relative abundance varied among treatments, as indicated by changes in 16S rRNA and 28S rRNA genes; (iv) changes in precipitation altered the relative abundance of Proteobacteria and Acidobacteria, where Acidobacteria decreased with a concomitant increase in the Proteobacteria in wet relative to dry treatments; and (v) changes in precipitation altered fungal community composition, primarily through lineage specific changes within a recently discovered group known as soil clone group I. Taken together, our results indicate that climate change drivers and their interactions may cause changes in bacterial and fungal overall abundance; however, changes in precipitation tended to have a much greater effect on the community composition. These results illustrate the potential for complex community changes in terrestrial ecosystems under climate change scenarios that alter multiple factors simultaneously. PMID- 20023090 TI - Functional role of Bradyrhizobium japonicum trehalose biosynthesis and metabolism genes during physiological stress and nodulation. AB - Trehalose, a disaccharide accumulated by many microorganisms, acts as a protectant during periods of physiological stress, such as salinity and desiccation. Previous studies reported that the trehalose biosynthetic genes (otsA, treS, and treY) in Bradyrhizobium japonicum were induced by salinity and desiccation stresses. Functional mutational analyses indicated that disruption of otsA decreased trehalose accumulation in cells and that an otsA treY double mutant accumulated an extremely low level of trehalose. In contrast, trehalose accumulated to a greater extent in a treS mutant, and maltose levels decreased relative to that seen with the wild-type strain. Mutant strains lacking the OtsA pathway, including the single, double, and triple DeltaotsA, DeltaotsA DeltatreS and DeltaotsA DeltatreY, and DeltaotsA DeltatreS DeltatreY mutants, were inhibited for growth on 60 mM NaCl. While mutants lacking functional OtsAB and TreYZ pathways failed to grow on complex medium containing 60 mM NaCl, there was no difference in the viability of the double mutant strain when cells were grown under conditions of desiccation stress. In contrast, mutants lacking a functional TreS pathway were less tolerant of desiccation stress than the wild-type strain. Soybean plants inoculated with mutants lacking the OtsAB and TreYZ pathways produced fewer mature nodules and a greater number of immature nodules relative to those produced by the wild-type strain. Taken together, results of these studies indicate that stress-induced trehalose biosynthesis in B. japonicum is due mainly to the OtsAB pathway and that the TreS pathway is likely involved in the degradation of trehalose to maltose. Trehalose accumulation in B. japonicum enhances survival under conditions of salinity stress and plays a role in the development of symbiotic nitrogen-fixing root nodules on soybean plants. PMID- 20023091 TI - Direct-imaging-based quantification of Bacillus cereus ATCC 14579 population heterogeneity at a low incubation temperature. AB - Bacillus cereus ATCC 14579 was cultured in microcolonies on Anopore strips near its minimum growth temperature to directly image and quantify its population heterogeneity at an abusive refrigeration temperature. Eleven percent of the microcolonies failed to grow during low-temperature incubation, and this cold induced population heterogeneity could be partly attributed to the loss of membrane integrity of individual cells. PMID- 20023092 TI - Inhibitory effects of low-energy pulsed ultrasonic stimulation on cell surface protein antigen C through heat shock proteins GroEL and DnaK in Streptococcus mutans. AB - This study concerns the use of low-energy pulsed ultrasound as nondestructive photodynamic antimicrobial therapy for controlling dental plaque. We examined the antibacterial and bactericidal effects of low-energy pulsed ultrasound on mutans streptococci and its inhibitory effects on bacterial cell adhesion of Streptococcus mutans. The results indicated weak antibacterial and bactericidal effects. However, ultrasonic stimulation for less than 20 min markedly decreased bacterial cell adhesion. To analyze the mechanism underlying the inhibitory effect, we examined cell surface protein antigen C (PAc) and glucosyltransferase I (GTF-I) expression in S. mutans. The levels of PAc gene and protein expression were markedly decreased by ultrasonic stimulation for 20 min. However, no change in GTF-I expression was observed. The expression of stress response heat shock proteins GroEL and DnaK was also examined. GroEL and DnaK levels were significantly decreased by ultrasonic stimulation, and the expression of the PAc protein was also diminished upon the addition of GroEL or DnaK inhibitors without ultrasonic stimulation. These observations suggest that the expression of the PAc protein in S. mutans may be dependent on heat shock proteins. Thus, low-energy pulsed ultrasound decreases bacterial adhesion by the inhibitory effect on the PAc protein and heat shock protein expression and may be useful as photodynamic antimicrobial chemotherapy in controlling dental plaque. PMID- 20023093 TI - High heterogeneity within methicillin-resistant Staphylococcus aureus ST398 isolates, defined by Cfr9I macrorestriction-pulsed-field gel electrophoresis profiles and spa and SCCmec types. AB - During recent years, the animal-associated methicillin-resistant Staphylococcus aureus clone ST398 has extensively been studied. The DNA of these isolates turned out to be refractory to SmaI restriction, and consequently, SmaI is unsuitable for subtyping this clone by standard pulsed-field gel electrophoresis (PFGE). Very recently, ST398 DNA was shown to be digested by Cfr9I, a neoschizomer of SmaI. In the present study, we employed Cfr9I PFGE on 100 German and 5 Dutch ST398 isolates and compared their PFGE profiles, protein A gene variable repeat regions (spa types), and types of the staphylococcal cassette chromosome mec (SCCmec). The isolates (from healthy carrier pigs, clinical samples from pigs, dust from farms, milk, and meat) were assigned to 35 profiles, which were correlated to the SCCmec type. A dendrogram with the Cfr9I patterns assigned all profiles to two clusters. Cluster A grouped nearly all isolates with SCCmec type V, and cluster B comprised all SCCmec type IVa and V* (a type V variant first identified as III) carriers plus one isolate with SCCmec type V. Both clusters also grouped methicillin-susceptible S. aureus isolates. The association of the majority of isolates with SCCmec type V in one large cluster indicated the presence of a successful subclone within the clonal complex CC398 from pigs, which has diversified. In general, the combination of Cfr9I PFGE with spa and SCCmec typing demonstrated the heterogeneity of the series analyzed and can be further used for outbreak investigations and traceability studies of the MRSA ST398 emerging clone. PMID- 20023094 TI - Transport and distribution of Salmonella enterica serovar Typhimurium in loamy and sandy soil monoliths with applied liquid manure. AB - A leaching experiment, where liquid manure spiked with Salmonella enterica serovar Typhimurium (Tet(+)) DSM554 was applied to soil surfaces, was conducted on intact soil monoliths (60 cm in diameter and 100 cm long). A total of 6.5 x 10(10) CFU was applied to each column. We found that Salmonella serovar Typhimurium could be transported to a 1-m depth in loamy soil at concentrations reaching 1.3 x 10(5) CFU/ml of leachate. The test strain was found in concentrations ranging from 300 to 1.3(5) cells/ml in loamy soil throughout the 27 days of the experiment, while concentrations below 20 cells/ml were sporadically detected in the leachates from sandy monoliths. Real-time PCR targeting invA DNA showed a clear correspondence between the total and culturable numbers of cells in the leachate, indicating that most cells leached were viable. On day 28, distribution of Salmonella serovar Typhimurium at five depths in the four monoliths was determined. The highest recovery rate, ranging from 1.5% to 3.8% of the total applied inoculum, was found in the top 0.2 m. PMID- 20023095 TI - Effect of daily temperature fluctuation during the cool season on the infectivity of Cryptosporidium parvum. AB - The present work calculated the rate of inactivation of Cryptosporidium parvum oocysts attributable to daily oscillations of low ambient temperatures. The relationship between air temperature and the internal temperature of bovine feces on commercial operations was measured, and three representative 24-h thermal regimens in the approximately 15 degrees C, approximately 25 degrees C, and approximately 35 degrees C ranges were chosen and emulated using a thermocycler. C. parvum oocysts suspended in deionized water were exposed to the temperature cycles, and their infectivity in mice was tested. Oral inoculation of 10(3) treated oocysts per neonatal BALB/c mouse (approximately 14 times the 50% infective dose) resulted in time- and temperature-dependent reductions in the proportion of infected mice. Oocysts were completely noninfectious after 14 24-h cycles with the 30 degrees C regimen and after 70 24-h cycles with the 20 degrees C regimen. In contrast, oocysts remained infectious after 90 24-h cycles with the 10 degrees C regimens. The estimated numbers of days needed for a 1-log(10) reduction in C. parvum oocyst infectivity were 4.9, 28.7, and 71.5 days for the 30, 20, and 10 degrees C thermal regimens, respectively. The loss of infectivity of oocysts induced by these thermal regimens was due in part to partial or complete in vitro excystation. PMID- 20023096 TI - Regulation of arabinose and xylose metabolism in Escherichia coli. AB - Bacteria such as Escherichia coli will often consume one sugar at a time when fed multiple sugars, in a process known as carbon catabolite repression. The classic example involves glucose and lactose, where E. coli will first consume glucose, and only when it has consumed all of the glucose will it begin to consume lactose. In addition to that of lactose, glucose also represses the consumption of many other sugars, including arabinose and xylose. In this work, we characterized a second hierarchy in E. coli, that between arabinose and xylose. We show that, when grown in a mixture of the two pentoses, E. coli will consume arabinose before it consumes xylose. Consistent with a mechanism involving catabolite repression, the expression of the xylose metabolic genes is repressed in the presence of arabinose. We found that this repression is AraC dependent and involves a mechanism where arabinose-bound AraC binds to the xylose promoters and represses gene expression. Collectively, these results demonstrate that sugar utilization in E. coli involves multiple layers of regulation, where cells will consume first glucose, then arabinose, and finally xylose. These results may be pertinent in the metabolic engineering of E. coli strains capable of producing chemical and biofuels from mixtures of hexose and pentose sugars derived from plant biomass. PMID- 20023097 TI - Physical characteristics of spores of food-associated isolates of the Bacillus cereus group. AB - All 47 food-borne isolates of Bacillus cereus sensu stricto, as well as 10 of 12 food-borne, enterotoxigenic isolates of Bacillus thuringiensis, possessed appendages. Spores were moderately to highly hydrophobic, and each had a net negative charge. These characteristics indicate that spores of food-associated B. thuringiensis and not only B. cereus sensu stricto have high potential to adhere to inert surfaces. PMID- 20023098 TI - Prolonged survival of Campylobacter species in bovine manure compost. AB - The persistence of naturally occurring campylobacteria in aerobic compost constructed of manure from beef cattle that were administered chlortetracycline and sulfamethazine (AS700) or from cattle not administered antibiotics (control) was examined. Although there were no differences in population sizes of heterotrophic bacteria, the temperature of AS700 compost was more variable and did not become as high as that of control compost. There were significant differences in water content, total carbon (C), total nitrogen (N), and electrical conductivity but not in the C/N ratio or pH between the two compost treatments. Campylobacteria were readily isolated from pen manure, for up to day 15 from control compost, and throughout the active phase of AS700 compost. Campylobacter DNA (including Campylobacter coli, Campylobacter fetus, Campylobacter hyointestinalis, and Campylobacter jejuni) was detected over the ca. 10-month composting period, and no reductions in quantities of C. jejuni DNA were observed over the duration of the active phase. The utilization of centrifugation in combination with ethidium monoazide (EMA) significantly reduced (>90%) the amplification of C. jejuni DNA that did not originate from cells with intact cell membranes. No differences were observed in the frequency of Campylobacter DNA detection between EMA- and non-EMA-treated samples, suggesting that Campylobacter DNA amplified from compost was extracted from cells with intact cell membranes (i.e., from viable cells). The findings of this study indicate that campylobacteria excreted in cattle feces persist for long periods in compost and call into question the common belief that these bacteria do not persist in manure. PMID- 20023099 TI - Mechanisms involved in governing adherence of Vibrio cholerae to granular starch. AB - Vibrio cholerae has been shown to adhere to cornstarch granules. The present work explored the mechanisms involved in this adhesion and the possibility of its occurrence in vivo. The findings suggest that both specific and nonspecific interactions are involved in the adhesion. Nonspecific hydrophobic interactions may play a role, since both V. cholerae and cornstarch granules exhibited hydrophobic properties when they were tested using a xylene-water system. In addition, the presence of bile acids reduced the adhesion. The adhesion also involves some specific carbohydrate-binding moieties on the cell surface, as reflected by reduced adhesion following pretreatment of the bacteria with proteinase K and sodium m-periodate. Further investigations supported these observations and showed that media containing low-molecular-weight carbohydrates had a significant inhibitory effect. Binding cell lysate to starch granules and removing the adhered proteins using either glucose or bile acids led to identification (by liquid chromatography-tandem mass spectrometry analysis) of several candidate V. cholerae outer membrane-associated starch-binding proteins. Different sets of proteins were isolated by removal in a glucose solution or bile acids. When the upper gastrointestinal tract conditions were simulated in vitro, both bile salts and the amylolytic activity of the pancreatic juices were found to have an inhibitory effect on the adherence of V. cholerae to starch. However, during acute diarrhea, this inhibitory effect may be significantly reduced due to dilution, suggesting that adhesion does occur in vivo. Such adhesion may contribute to the beneficial effects observed following administration of granular starch-based oral rehydration solutions to cholera patients. PMID- 20023100 TI - Seasonal retention and release of Cryptosporidium parvum oocysts by environmental biofilms in the laboratory. AB - Cryptosporidium is a genus of waterborne protozoan parasites that causes significant gastrointestinal disease in humans. These parasites can accumulate in environmental biofilms and be subsequently released to contaminate water supplies. Natural microbial assemblages were collected each season from an eastern Pennsylvania stream and used to grow biofilms in laboratory microcosms in which influx, efflux, and biofilm retention were determined from daily oocyst counts. For each seasonal biofilm, oocysts attached to the biofilm quickly during oocyst dosing. Upon termination of oocyst dosing, the percentage of oocysts retained within the biofilm decreased to a new steady state within 5 days. Seasonal differences in biofilm retention of oocysts were observed. The spring biofilm retained the greatest percentage of oocysts, followed (in decreasing order) by the winter, summer, and fall biofilms. There was no statistically significant correlation between the percentage of oocysts attached to the biofilm and (i) any measured stream water quality parameter (including temperature, pH, conductivity, and dissolved organic carbon concentration) or (ii) experimental temperature. Seasonal differences in oocyst retention persisted when biofilms were tested with stream water from a different season. These data suggest that seasonal variation in the microbial community and resulting biofilm architecture may be more important to oocyst transport in this stream site than water quality. The biofilm attachment and detachment dynamics of C. parvum oocysts have implications for public health, and the drinking water industry should recognize that the potential exists for pathogen-free water to become contaminated during the distribution process as a result of biofilm dynamics. PMID- 20023101 TI - Use of a foam spatula for sampling surfaces after bioaerosol deposition. AB - The present study had three goals: (i) to evaluate the relative quantities of aerosolized Bacillus atrophaeus spores deposited on the vertical, horizontal top, and horizontal bottom surfaces in a chamber; (ii) to assess the relative recoveries of the aerosolized spores from glass and stainless steel surfaces with a polyester swab and a macrofoam sponge wipe; and (iii) to estimate the relative recovery efficiencies of aerosolized B. atrophaeus spores and Pantoea agglomerans using a foam spatula at several different bacterial loads by aerosol distribution on glass surfaces. The majority of spores were collected from the bottom horizontal surface regardless of which swab type and extraction protocol were used. Swabbing with a macrofoam sponge wipe was more efficient in recovering spores from surfaces contaminated with high bioaerosol concentrations than swabbing with a polyester swab. B. atrophaeus spores and P. agglomerans culturable cells were detected on glass surfaces using foam spatulas when the theoretical surface bacterial loads were 2.88 x 10(4) CFU and 8.09 x 10(6) CFU per 100-cm(2) area, respectively. The median recovery efficiency from the surfaces using foam spatulas was equal to 9.9% for B. atrophaeus spores when the recovery was calculated relative to the theoretical surface spore load. Using a foam spatula permits reliable sampling of spores on the bioaerosol-exposed surfaces in a wide measuring range. The culturable P. agglomerans cells were recovered with a median efficiency of 0.001%, but staining the swab extracts with fluorescent dyes allowed us to observe that the viable cell numbers were higher by 1.83 log units than culturable organisms. However, additional work is needed to improve the analysis of the foam extracts in order to decrease the limit of detection of Bacillus spores and Gram-negative bacteria on contaminated surfaces. PMID- 20023102 TI - Yeast surface display of trifunctional minicellulosomes for simultaneous saccharification and fermentation of cellulose to ethanol. AB - By combining cellulase production, cellulose hydrolysis, and sugar fermentation into a single step, consolidated bioprocessing (CBP) represents a promising technology for biofuel production. Here we report engineering of Saccharomyces cerevisiae strains displaying a series of uni-, bi-, and trifunctional minicellulosomes. These minicellulosomes consist of (i) a miniscaffoldin containing a cellulose-binding domain and three cohesin modules, which was tethered to the cell surface through the yeast a-agglutinin adhesion receptor, and (ii) up to three types of cellulases, an endoglucanase, a cellobiohydrolase, and a beta-glucosidase, each bearing a C-terminal dockerin. Cell surface assembly of the minicellulosomes was dependent on expression of the miniscaffoldin, indicating that formation of the complex was dictated by the high-affinity interactions between cohesins and dockerins. Compared to the unifunctional and bifunctional minicellulosomes, the quaternary trifunctional complexes showed enhanced enzyme-enzyme synergy and enzyme proximity synergy. More importantly, surface display of the trifunctional minicellulosomes gave yeast cells the ability to simultaneously break down and ferment phosphoric acid-swollen cellulose to ethanol with a titer of approximately 1.8 g/liter. To our knowledge, this is the first report of a recombinant yeast strain capable of producing cell associated trifunctional minicellulosomes. The strain reported here represents a useful engineering platform for developing CBP-enabling microorganisms and elucidating principles of cellulosome construction and mode of action. PMID- 20023103 TI - Comparison of PCR binary typing (P-BIT), a new approach to epidemiological subtyping of Campylobacter jejuni, with serotyping, pulsed-field gel electrophoresis, and multilocus sequence typing methods. AB - To overcome some of the deficiencies with current molecular typing schema for Campylobacter spp., we developed a prototype PCR binary typing (P-BIT) approach. We investigated the distribution of 68 gene targets in 58 Campylobacter jejuni strains, one Campylobacter lari strain, and two Campylobacter coli strains for this purpose. Gene targets were selected on the basis of distribution in multiple genomes or plasmids, and known or putative status as an epidemicity factor. Strains were examined with Penner serotyping, pulsed-field gel electrophoresis (PFGE; using SmaI and KpnI enzymes), and multilocus sequence typing (MLST) approaches for comparison. P-BIT provided 100% typeability for strains and gave a diversity index of 98.5%, compared with 97.0% for SmaI PFGE, 99.4% for KpnI PFGE, 96.1% for MLST, and 92.8% for serotyping. Numerical analysis of the P-BIT data clearly distinguished strains of the three Campylobacter species examined and correlated somewhat with MLST clonal complex assignations and with previous classifications of "high" and "low" risk. We identified 18 gene targets that conferred the same level of discrimination as the 68 initially examined. We conclude that P-BIT is a useful approach for subtyping, offering advantages of speed, cost, and potential for strain risk ranking unavailable from current molecular typing schema for Campylobacter spp. PMID- 20023104 TI - Composition of the landfill microbial community as determined by application of domain- and group-specific 16S and 18S rRNA-targeted oligonucleotide probes. AB - The microbial community composition of colonized cotton and leachate samples from a landfill was quantified using small subunit (SSU) rRNA probes (quantitative rRNA hybridization). Relative quantification of bacteria, eukaryotes, and archaea revealed variations in the landfill microbial community between samples from different areas of the landfill site and indicated the presence of potentially novel archaea. Anaerobic fungi were quantified in rumen fluid samples but were not sufficiently abundant for direct detection in the landfill samples. PMID- 20023105 TI - Adaptation of the highly productive T7 expression system to Streptomyces lividans. AB - Streptomyces lividans is a Gram-positive bacterium known for its remarkable secretion efficiency and low extracellular protease activity. In the present work, we adapted the highly productive T7 expression system to S. lividans. A codon-optimized T7 RNA polymerase gene was chromosomally integrated, and a bifunctional T7 expression vector was constructed. PMID- 20023106 TI - Metabolic impact of increased NADH availability in Saccharomyces cerevisiae. AB - Engineering the level of metabolic cofactors to manipulate metabolic flux is emerging as an attractive strategy for bioprocess applications. We present the metabolic consequences of increasing NADH in the cytosol and the mitochondria of Saccharomyces cerevisiae. In a strain that was disabled in formate metabolism, we either overexpressed the native NAD(+)-dependent formate dehydrogenase in the cytosol or directed it into the mitochondria by fusing it with the mitochondrial signal sequence encoded by the CYB2 gene. Upon exposure to formate, the mutant strains readily consumed formate and induced fermentative metabolism even under conditions of glucose derepression. Cytosolic overexpression of formate dehydrogenase resulted in the production of glycerol, while when this enzyme was directed into the mitochondria, we observed glycerol and ethanol production. Clearly, these results point toward different patterns of compartmental regulation of redox homeostasis. When pulsed with formate, S. cerevisiae cells growing in a steady state on glucose immediately consumed formate. However, formate consumption ceased after 20 min. Our analysis revealed that metabolites at key branch points of metabolic pathways were affected the most by the genetic perturbations and that the intracellular concentrations of sugar phosphates were specifically affected by time. In conclusion, the results have implications for the design of metabolic networks in yeast for industrial applications. PMID- 20023107 TI - Caldicellulosiruptor obsidiansis sp. nov., an anaerobic, extremely thermophilic, cellulolytic bacterium isolated from Obsidian Pool, Yellowstone National Park. AB - A novel, obligately anaerobic, extremely thermophilic, cellulolytic bacterium, designated OB47(T), was isolated from Obsidian Pool, Yellowstone National Park, WY. The isolate was a nonmotile, non-spore-forming, Gram-positive rod approximately 2 microm long by 0.2 microm wide and grew at temperatures between 55 and 85 degrees C, with the optimum at 78 degrees C. The pH range for growth was 6.0 to 8.0, with values of near 7.0 being optimal. Growth on cellobiose produced the fastest specific growth rate at 0.75 h(-1). The organism also displayed fermentative growth on glucose, maltose, arabinose, fructose, starch, lactose, mannose, sucrose, galactose, xylose, arabinogalactan, Avicel, xylan, filter paper, processed cardboard, pectin, dilute acid-pretreated switchgrass, and Populus. OB47(T) was unable to grow on mannitol, fucose, lignin, Gelrite, acetate, glycerol, ribose, sorbitol, carboxymethylcellulose, and casein. Yeast extract stimulated growth, and thiosulfate, sulfate, nitrate, and sulfur were not reduced. Fermentation end products were mainly acetate, H2, and CO2, although lactate and ethanol were produced in 5-liter batch fermentations. The G+C content of the DNA was 35 mol%, and sequence analysis of the small subunit rRNA gene placed OB47(T) within the genus Caldicellulosiruptor. Based on its phylogenetic and phenotypic properties, the isolate is proposed to be designated Caldicellulosiruptor obsidiansis sp. nov. and OB47 is the type strain (ATCC BAA 2073). PMID- 20023108 TI - Association of hemolytic activity of Pseudomonas entomophila, a versatile soil bacterium, with cyclic lipopeptide production. AB - Pseudomonas entomophila is an entomopathogenic bacterium that is able to infect and kill Drosophila melanogaster upon ingestion. Its genome sequence suggests that it is a versatile soil bacterium closely related to Pseudomonas putida. The GacS/GacA two-component system plays a key role in P. entomophila pathogenicity, controlling many putative virulence factors and AprA, a secreted protease important to escape the fly immune response. P. entomophila secretes a strong diffusible hemolytic activity. Here, we showed that this activity is linked to the production of a new cyclic lipopeptide containing 14 amino acids and a 3 C(10)OH fatty acid that we called entolysin. Three nonribosomal peptide synthetases (EtlA, EtlB, EtlC) were identified as responsible for entolysin biosynthesis. Two additional components (EtlR, MacAB) are necessary for its production and secretion. The P. entomophila GacS/GacA two-component system regulates entolysin production, and we demonstrated that its functioning requires two small RNAs and two RsmA-like proteins. Finally, entolysin is required for swarming motility, as described for other lipopeptides, but it does not participate in the virulence of P. entomophila for Drosophila. While investigating the physiological role of entolysin, we also uncovered new phenotypes associated with P. entomophila, including strong biocontrol abilities. PMID- 20023110 TI - Critical evaluation of the volumetric "bottle effect" on microbial batch growth. AB - We have analyzed the impact of surface-to-volume ratio on final bacterial concentrations after batch growth. We examined six bottle sizes (20 to 1,000 ml) using three independent enumeration methods to quantify growth. We found no evidence of a so-called volumetric bottle effect, thus contradicting numerous previous reports. PMID- 20023109 TI - Musca domestica salivary gland hypertrophy virus, a globally distributed insect virus that infects and sterilizes female houseflies. AB - The housefly, Musca domestica, is a cosmopolitan pest of livestock and poultry and is of economic, veterinary, and public health importance. Populations of M. domestica are naturally infected with M. domestica salivary gland hypertrophy virus (MdSGHV), a nonoccluded double-stranded DNA virus that inhibits egg production in infected females and is characterized by salivary gland hypertrophy (SGH) symptoms. MdSGHV has been detected in housefly samples from North America, Europe, Asia, the Caribbean, and the southwestern Pacific. In this study, houseflies were collected from various locations and dissected to observe SGH symptoms, and infected gland pairs were collected for MdSGHV isolation and amplification in laboratory-reared houseflies. Differences among the MdSGHV isolates were examined by using molecular and bioassay approaches. Approximately 600-bp nucleotide sequences from each of five open reading frames having homology to genes encoding DNA polymerase and partial homology to the genes encoding four per os infectivity factor proteins (p74, pif-1, pif-2, and pif-3) were selected for phylogenetic analyses. Nucleotide sequences from 16 different geographic isolates were highly homologous, and the polymorphism detected was correlated with geographic source. The virulence of the geographic MdSGHV isolates was evaluated by per os treatment of newly emerged and 24-h-old houseflies with homogenates of infected salivary glands. In all cases, 24-h-old flies displayed a resistance to oral infection that was significantly greater than that displayed by newly eclosed adults. Regardless of the MdSGHV isolate tested, all susceptible insects displayed similar degrees of SGH and complete suppression of oogenesis. PMID- 20023111 TI - CalA, a cyanobacterial AbrB protein, interacts with the upstream region of hypC and acts as a repressor of its transcription in the cyanobacterium Nostoc sp. strain PCC 7120. AB - The filamentous, heterocystous, nitrogen-fixing cyanobacterium Nostoc sp. strain PCC 7120 may contain, depending on growth conditions, up to two hydrogenases directly involved in hydrogen metabolism. HypC is one out of at least seven auxiliary gene products required for synthesis of a functional hydrogenase, specifically involved in the maturation of the large subunit. In this study we present a protein, CalA (Alr0946 in the genome), belonging to the transcription regulator family AbrB, which in protein-DNA assays was found to interact with the upstream region of hypC. Transcriptional investigations showed that calA is cotranscribed with the downstream gene alr0947, which encodes a putative protease from the abortive infection superfamily, Abi. CalA was shown to interact specifically not only with the upstream region of hypC but also with its own upstream region, acting as a repressor on hypC. The bidirectional hydrogenase activity was significantly downregulated when CalA was overexpressed, demonstrating a correlation with the transcription factor, either direct or indirect. In silico studies showed that homologues to both CalA and Alr0947 are highly conserved proteins within cyanobacteria with very similar physical organizations of the corresponding structural genes. Possible functions of the cotranscribed downstream protein Alr0947 are presented. In addition, we present a three-dimensional (3D) model of the DNA binding domain of CalA and putative DNA binding mechanisms are discussed. PMID- 20023114 TI - Translumbar central venous catheters for long-term haemodialysis. AB - BACKGROUND: Vascular access for haemodialysis is achieved by tunnelled central venous catheter (CVC) in at least 23% of prevalent patients in the UK, Canada and the USA. Use of CVCs is associated with an increased incidence of venous stenosis that can progressively limit future vascular access routes. Lack of conventional venous access routes mandates the use of alternative strategies such as the translumbar approach. METHODS: We retrospectively analysed patients at our centre requiring translumbar inferior vena caval CVCs (TesioCath) for haemodialysis in the period 1999-2008. Written and electronic records capturing dialysis adequacy and complications, hospital admissions and laboratory data were examined. RESULTS: Thirty-nine pairs of translumbar CVCs were inserted in 26 patients with 15 864 catheter days follow-up, mean patient age 61.9 +/- 12.1 years, 31% diabetic, 15% with ischaemic heart disease. All insertions were successful. Insertion of one CVC was associated with a self-limiting retroperitoneal haematoma. No patients died of a catheter-related cause or through lack of vascular access. Cumulative assisted primary catheter site patency was 81% at 6 months and 73% at 1 year (median 18.5 months). Good dialysis adequacy was achieved throughout (mean single-pool Kt/V 1.5 +/- 0.4). The incidence of access related infection was 2.84/1000 catheter days (exit site infection rate 2.02/1000 catheter days; catheter-related bacteraemia rate 0.82/1000 catheter days). Catheter dysfunction (need for thrombolytic infusion or catheter change) led to 0.88 admissions per 1000 catheter days. CONCLUSION: Translumbar inferior vena caval CVCs can offer relatively safe and effective long-term haemodialysis access in patients with no other options. PMID- 20023118 TI - Diabetic conditions act as matchmaker for monocytes and vascular smooth muscle cells. PMID- 20023120 TI - Divergent roles of prokineticin receptors in the endothelial cells: angiogenesis and fenestration. AB - Prokineticins are secreted peptides that activate two G protein-coupled receptors: PKR1 and PKR2. Prokineticins induce angiogenesis and fenestration, but the cognate receptors involved in these functions are unknown. We hypothesized a role for prokineticin receptor signaling pathways and expression profiles in determining the selective effects of prokineticins on coronary endothelial cells (H5V). Activation of the PKR1/MAPK/Akt signaling pathway stimulates proliferation, migration, and angiogenesis in H5V cells, in which PKR1 predominates over PKR2. PKR1 was colocalized with Galpha(11) and was internalized following the stimulation of these cells with prokineticin-2. Knock down of PKR1 or Galpha(11) expression in H5V cells effectively inhibited prokineticin-2 induced vessel formation and MAPK/Akt activation, indicating a role for PKR1/Galpha(11) in this process. However, in conditions in which PKR2 predominated over PKR1, these cells displayed a fenestrated endothelial cell phenotype. H5V cells overexpressing PKR2 displayed large numbers of multivesicular bodies and caveolar clusters and a disruption of the distribution of zonula occluden-1 tight junction protein. Prokineticin-2 induced the colocalization of PKR2 with Galpha(12), and activated Galpha(12), which bound to zonula occluden-1 to trigger the degradation of this protein in these cells. Prokineticin-2 induced the formation of vessel-like structures by human aortic endothelial cells expressing only PKR1, and disorganized the tight junctions in human hepatic sinusoidal endothelial cells expressing only PKR2, confirming the divergent roles of these receptors. Our findings show the functional characteristics of coronary endothelial cells depend on the expression of PKR1 and PKR2 levels and the divergent signaling pathways used by these receptors. PMID- 20023119 TI - Calmodulin kinase II is required for angiotensin II-mediated vascular smooth muscle hypertrophy. AB - Despite our understanding that medial smooth muscle hypertrophy is a central feature of vascular remodeling, the molecular pathways underlying this pathology are still not well understood. Work over the past decade has illustrated a potential role for the multifunctional calmodulin-dependent kinase CaMKII in smooth muscle cell contraction, growth, and migration. Here we demonstrate that CaMKII is enriched in vascular smooth muscle (VSM) and that CaMKII inhibition blocks ANG II-dependent VSM cell hypertrophy in vitro and in vivo. Specifically, systemic CaMKII inhibition with KN-93 prevented ANG II-mediated hypertension and medial hypertrophy in vivo. Adenoviral transduction with the CaMKII peptide inhibitor CaMKIIN abrogated ANG II-induced VSM hypertrophy in vitro, which was augmented by overexpression of CaMKII-delta2. Finally, we identify the downstream signaling components critical for ANG II- and CaMKII-mediated VSM hypertrophy. Specifically, we demonstrate that CaMKII induces VSM hypertrophy by regulating histone deacetylase 4 (HDAC4) activity, thereby stimulating activity of the hypertrophic transcription factor MEF2. MEF2 transcription is activated by ANG II in vivo and abrogated by the CaMKII inhibitor KN-93. Together, our studies identify a complete pathway for ANG II-triggered arterial VSM hypertrophy and identify new potential therapeutic targets for chronic human hypertension. PMID- 20023121 TI - Physical activity and heart rate variability measured simultaneously during waking hours. AB - Heart rate (HR) variability (HRV) during ambulatory recordings may be affected by individual differences in daily physical activity (PA). However, the influence of various levels of PA on different measures of HRV is not exactly known. We examined the association between simultaneously measured HRV and objective PA data obtained with an accelerometer during waking hours among 45 healthy adults. Bouts of PA were identified from minute-by-minute accelerometer data as metabolic equivalent (METs) values and calculated as mean METs for 30 min. HRV was analyzed concurrently. Within-individual correlation analyses and sign tests were performed to study the relationships between various HRV indexes and PA. The mean PA time was 15:44 +/- 1:01 h, and the mean MET was 1.91 +/- 0.14. HR and sample entropy, but not the other measures of HRV, had a significant relationship with PA, as shown by both correlation analyses (r = 0.64, P = 0.021, and r = -0.55, P = 0.022, respectively) and sign tests (P < 0.0001 for both). Beat-to-beat R-R interval fluctuation expressed as SD1 also demonstrated a significant relation to PA according to the sign test (P = 0.037) and a trend of association according to the correlation analysis (r = -0.40, P = 0.129). The complexity measure of HRV, in addition to average HR and the short-term index of HRV (SD1), is significantly influenced by the level of PA during ambulatory conditions. Long-term HRV indexes remained relatively stable at various activity levels, making them the most robust indexes for the assessment of cardiac autonomic function during free running ambulatory conditions. PMID- 20023122 TI - Mechanisms underlying hypothermia-induced cardiac contractile dysfunction. AB - Rewarming patients after profound hypothermia may result in acute heart failure and high mortality (50-80%). However, the underlying pathophysiological mechanisms are largely unknown. We characterized cardiac contractile function in the temperature range of 15-30 degrees C by measuring the intracellular Ca(2+) concentration ([Ca(2+)](i)) and twitch force in intact left ventricular rat papillary muscles. Muscle preparations were loaded with fura-2 AM and electrically stimulated during cooling at 15 degrees C for 1.5 h before being rewarmed to the baseline temperature of 30 degrees C. After hypothermia/rewarming, peak twitch force decreased by 30-40%, but [Ca(2+)](i) was not significantly altered. In addition, we assessed the maximal Ca(2+)-activated force (F(max)) and Ca(2+) sensitivity of force in skinned papillary muscle fibers. F(max) was decreased by approximately 30%, whereas the pCa required for 50% of F(max) was reduced by approximately 0.14. In rewarmed papillary muscle, both total cardiac troponin I (cTnI) phosphorylation and PKA-mediated cTnI phosphorylation at Ser23/24 were significantly increased compared with controls. We conclude that after hypothermia/rewarming, myocardial contractility is significantly reduced, as evidenced by reduced twitch force and F(max). The reduced myocardial contractility is attributed to decreased Ca(2+) sensitivity of force rather than [Ca(2+)](i) itself, resulting from increased cTnI phosphorylation. PMID- 20023124 TI - Cross-bridge cycling gives rise to spatiotemporal heterogeneity of dynamic subcellular mechanics in cardiac myocytes probed with atomic force microscopy. AB - To study how the dynamic subcellular mechanical properties of the heart relate to the fundamental underlying process of actin-myosin cross-bridge cycling, we developed a novel atomic force microscope elastography technique for mapping spatiotemporal stiffness of isolated, spontaneously beating neonatal rat cardiomyocytes. Cells were indented repeatedly at a rate close but unequal to their contractile frequency. The resultant changes in pointwise apparent elastic modulus cycled at a predictable envelope frequency between a systolic value of 26.2 +/- 5.1 kPa and a diastolic value of 7.8 +/- 4.1 kPa at a representative depth of 400 nm. In cells probed along their major axis, spatiotemporal changes in systolic stiffness displayed a heterogeneous pattern, reflecting the banded sarcomeric structure of underlying myofibrils. Treatment with blebbistatin eliminated contractile activity and resulted in a uniform apparent modulus of 6.5 +/- 4.8 kPa. This study represents the first quantitative dynamic mechanical mapping of beating cardiomyocytes. The technique provides a means of probing the micromechanical effects of disease processes and pharmacological treatments on beating cardiomyocytes, providing new insights and relating subcellular cardiac structure and function. PMID- 20023123 TI - Uterine and fetal blood flow indexes and fetal growth assessment after chronic estrogen suppression in the second half of baboon pregnancy. AB - Although estrogen regulates important aspects of maternal cardiovascular physiology, the role of estrogen on uteroplacental and fetal blood flow is incompletely understood. This study tested the hypothesis that chronically suppressing endogenous estrogen production during the second half of baboon pregnancy alters uterine and fetal blood flow dynamics assessed by ultrasonography. Pregnant baboons were untreated or treated daily with the aromatase inhibitor letrozole or letrozole plus estradiol on days 100-160 of gestation (term = 184 days). Blood flow dynamics were determined by Doppler ultrasonography on day 60 and longitudinally between days 110 and 160 of gestation. Letrozole decreased maternal serum estradiol and estrone concentrations by 95% (P < 0.001). Fetal growth biometrical parameters increased (P < 0.001) between days 110 and 160 of gestation and were similar in untreated and letrozole-treated animals. Uterine, umbilical, and fetal middle cerebral artery pulsatility index and resistance index declined (P < 0.01) by 30-50% and uterine artery volume flow increased sixfold (P < 0.001) between days 60 and 160, but values were similar in untreated, letrozole-treated, and letrozole plus estradiol-treated baboons. Thus uterine and fetal artery blood flow indexes, uterine artery volume flow, and fetal growth were maintained at normal levels despite chronic estrogen suppression in the second half of baboon pregnancy. This suggests that elevated levels of endogenous estrogen are not required to maintain low impedance blood flow within the uteroplacental vascular bed during the second half of nonhuman primate pregnancy. PMID- 20023125 TI - Pericytes in the macrovascular intima: possible physiological and pathogenetic impact. AB - The frequently observed de-endothelialization of venous coronary bypass grafts prepared using standard methods exposes subendothelial prothrombotic cells to blood components, thus endangering patients by inducing acute thromboembolic infarction or long-term proliferative stenosis. Our aim was to gain deeper histological and physiological insight into these relations. An intricate network of subendothelial cells, characterized by histological features specific for true pericytes, was detected even in healthy vessels and forms, coupled to the luminal endothelium, a second leaflet of the macrovascular intima. These cells, and particularly those in the venous intima, express enormous concentrations of tissue factor and can recruit additional amounts of up to the 25-fold concentration within 1 h during preincubation with serum (intimal pericytes of venous origin activate 30.71 +/- 4.07 pmol coagulation factor x.min(-1).10(-6) cells; n = 15). Moreover, decoupled from the endothelium, they proliferate rapidly (generation time, 15 +/- 2.1 h, n = 8). Central regions of atherosclerotic plaques, as well as of those of restenosed areas of coronary vein grafts, consist almost completely of these cells. In stark contrast with the prothrombogenicity of the intimal pericytes, intact luminal endothelium recruits high concentrations of thrombomodulin (CD 141) specifically within its intercellular junctions, activates Protein C rapidly (42 +/- 5.1 pmol/min.10(6) venous endothelial cells at thrombin saturation; n = 15), can thus actively prevent coagulatory processes, and never expresses histologically detectable and functionally active tissue factor. Given this strongly prothrombotic potential of the intimal pericytes and their overshooting growth behavior in endothelium denuded vascular regions, they may play important roles in the development of atherosclerosis, thrombosis, and saphenous vein graft disease. PMID- 20023126 TI - Energy and amino acid utilization in expeller-extracted canola meal fed to growing pigs. AB - Two experiments were conducted to determine the nutritive value of expeller extracted canola meal (EECM) for growing pigs. In Exp. 1, a total of 6 ileally cannulated barrows (average initial BW = 26.8 kg) were fed 3 diets in a replicated 3 x 3 Latin square design to determine the apparent and standardized ileal digestibility (SID) values of N and AA and the SID AA contents of EECM. The 3 diets were a cornstarch-based diet with either solvent-extracted canola meal (SECM) or EECM as the sole source of protein, and a low-casein cornstarch-based diet, which was used to estimate basal endogenous N and AA losses to determine the SID of N and AA. All 3 diets contained chromic oxide as an indigestible marker for determining nutrient digestibility by the indicator method. In Exp. 2, a total of 18 intact barrows (average initial BW = 25.9 kg) were fed 3 diets in a completely randomized design (6 pigs per diet) to determine apparent total tract digestibility and retention of nutrients and the DE and ME contents of EECM. The diets were a basal corn-based diet or the basal diet with corn replaced by 35% SECM or EECM. The basal diet was used for determining the total tract digestible nutrient content by the difference method. Solvent-extracted canola meal, which is commonly used in the formulation of swine diets, was fed in both experiments for comparison with EECM. The SECM and EECM were similar in CP content (41.8 vs. 41.4%). Expeller-extracted canola meal was, however, greater in ether extract content (12.03 vs. 5.54%) and decreased in NDF content (23.8 vs. 29.9%) compared with SECM. The EECM also had a greater content of all the AA except Met, Cys, and Ser, by approximately 6.6%; Cys was greater in SECM, whereas Met and Ser were similar between the 2 meals. The EECM had greater (P < 0.05) SID of N, Arg, Ile, Leu, Phe, Glu, and Pro. The SID contents of Arg, His, Ile, Leu, Phe, Val, Ala, Asp, Glu, Gly, Pro, and Tyr were also greater (P < 0.05) for EECM than for SECM by an average of 15%. The EECM had greater (P < 0.01) DE (4,107 vs. 3,790 kcal/kg) and ME (3,978 vs. 3,564 kcal/kg) values compared with SECM. The results show that the EECM used in the current study had greater digestible AA and energy and a greater ME content than the SECM; hence, it may be a better source of protein and energy for growing pigs than SECM. PMID- 20023127 TI - Introduction of meat sheep breeds in extensive systems of Patagonia: lamb growth and survival. AB - Genotype effects on lamb growth, survival, and commercial finishing were estimated in a 5-yr study aimed at assessing potential benefits from introducing meat breeds into extensive sheep systems of northeastern Patagonia. Five ram [Corriedale: CO; Border Leicester: BL; Ile de France: IF; Texel: TX; and synthetic CRIII (25% Merino, 37.5% IF, 37.5% TX)] and 5 dam (CO; synthetic CRIII; BLCO: BL x CO; IFCO: IF x CO; and TXCO: TX x CO) genotypes were represented in the study. Data were collected from 1,426 born and 1,258 weaned lambs of 9 resulting genotypes (CO x CO, BL x CO, IF x CO, TX x CO, CRIII x CO, CRIII x BLCO, CRIII x IFCO, CRIII x TXCO, and CRIII x CRIII). Birth weight was recorded on all lambs; subsequently, BW and BCS (scale 1 to 5) were recorded at regular intervals until weaning. Body weights were adjusted to 60 and 90 (weaning) d of age, and ADG were calculated from the adjusted BW for the periods 0 to 60 d and 60 to 90 d; BCS was adjusted to 90 d. Survival to weaning and percentage of lambs reaching commercial finishing (BW >or= 23 kg and BCS >or= 2.5 points) were also recorded. Significant (P < 0.05) genotype x litter size interactions were detected for birth weight and ADG 60 to 90 d. With the exception of CRIII x CO, crossbred and synthetic genotypes presented greater (P < 0.05) ADG 0 to 60 d and BW at 60 and 90 d than CO x CO lambs. Second cross lambs reared as singles presented greater ADG 60 to 90 d (P < 0.05) than BL x CO, TX x CO, and CRIII x CO, but less (P < 0.05) ADG 60 to 90 d, and no differences were observed for twins. The IF x CO, CRIII x BLCO, CRIII x IFCO, and CRIII x CRIII genotypes showed greater BCS at 90 d (P < 0.05) than CO x CO. The probability of greater commercial finishing for crossbred and synthetic genotypes relative to CO x CO was at least 79%. Probabilities of greater survival to weaning in CO x CO and CRIII x BLCO lambs relative to IF x CO, TX x CO, CRIII x CO, and CRIII x CRIII lambs were greater than 81%. Results indicate significant improvements in lamb BW and saleable lambs from the introduction of meat genotypes. Among the terminal sire breeds evaluated, BL and IF would produce the greatest impact. Differential nutritional management of pregnant ewes carrying twins and of twin lambs beyond 60 d should be implemented to mitigate litter size x genotype interactions constraining growth potential benefits, which may be critical for northeastern Patagonia conditions. PMID- 20023128 TI - Effect of beta-glucans contained in barley- and oat-based diets and exogenous enzyme supplementation on gastrointestinal fermentation of finisher pigs and subsequent manure odor and ammonia emissions. AB - The objective of the current experiment was to evaluate the influence of dietary cereal sources of beta(1,3)(1,4)-d-glucan (beta-glucan) and enzyme supplementation on indices of environmental pollution from finisher pigs. An experiment with a 2 x 2 factorial arrangement of treatments was initiated to investigate the effect of dietary source of beta-glucan (barley vs. oats) and enzyme supplementation (no vs. yes) on nutrient digestibility, N utilization, intestinal fermentation, and manure odor and ammonia emissions from finisher boars (n = 4; BW = 73.9 kg; SD = 4.7). Sixteen boars were assigned to 1 of 4 dietary treatments (n = 4/treatment): 1) barley-based diet, 2) barley-based diet + exogenous enzyme, 3) oat-based diet, and 4) oat-based diet + enzyme. The enzyme supplement used contained endo-1,3(4)-beta-glucanase and endo-1,4-beta-xylanase. Experimental diets were formulated to contain similar concentrations of DE (13.5 MJ/kg) and digestible lysine (8.8 g/kg). Pigs offered oat-based diets had reduced digestibility of DM (0.795 vs. 0.849; SEM 0.007; P = 0.001), OM (0.808 vs. 0.865; SEM 0.007; P = 0.001), GE (0.806 vs. 0.845; SEM 0.006; P = 0.002), and NDF (0.233 vs. 0.423; SEM 0.033; P < 0.003) compared with those offered barley-based diets. Oat-based diets increased populations of Bifidobacterium spp. (7.26 vs. 6.38 log cfu/g of digesta; SEM 0.201; P = 0.005) and Lactobacillus spp. (6.99 vs. 6.18 log cfu/g of digesta; SEM 0.234; P = 0.022) in the proximal colon and decreased manure odor emissions [2,179.6 vs. 4,984.6 Ou(E)/m(3) (where Ou(E) refers to European odor units); SEM 653.7; P < 0.011] compared with barley-based diets. There was an interaction between cereal type and enzyme inclusion on manure ammonia emissions from 0 to 96 h (P = 0.050). Pigs offered barley-based diets containing an enzyme supplement had increased manure ammonia emissions compared with those offered unsupplemented barley-based diets. However, there was no effect of enzyme inclusion on oat-based diets. In conclusion, pigs offered oat based diets harbored increased Bifidobacterium spp. and Lactobacillus spp. populations in the proximal colon and had decreased manure odor emissions compared with those offered barley-based diets. Enzyme inclusion had no effect on manure ammonia emissions from pigs offered oat-based diets. PMID- 20023129 TI - Comparative proteomic profiling of 2 muscles from 5 different pure pig breeds using surface-enhanced laser desorption/ionization time-of-flight proteomics technology. AB - The objectives of this study were to evaluate the influence of different pure pig breeds and muscle types on the expression of muscle proteins, as well as their interactions, and second, to find biomarkers for breed and muscle types. A total of 126 male pigs, including 43 Landrace, 21 Duroc, 43 Large White, 13 Pietrain, and 6 Belgian Landrace, were slaughtered at the age of 174 +/- 6 d. Samples from the semimembranosus muscle (SM) and LM were collected 24 h postmortem. Proteomic spectra were generated on an anion exchanger (Q10), a cation exchanger (CM10), and on immobilized metal affinity capture (IMAC30) ProteinChip arrays and analyzed using surface-enhanced laser desorption/ionization time-of-flight mass spectrometry ProteinChip techniques. Breed and muscle type did not affect the number of peaks per spectrum but, interestingly, affected the average intensity of the peaks. Of these peaks, a total of 4 proved to be potential protein biomarkers to differentiate LM or SM muscles, and 2 to classify specific breed types. Additionally, several peaks influenced by the interaction between muscle and breed types could correctly classify pig muscles according to their breed. Further studies need to be carried out to validate and identify these potential protein biomarkers for breed and muscle types in finishing pigs. PMID- 20023130 TI - Longitudinal changes in ultrasonic measurements of body composition during growth in Suffolk ram lambs and evaluation of alternative adjustment strategies for ultrasonic scan data. AB - Four equations were used to compare alternative procedures to adjust ultrasonic estimates (y) of backfat thickness (BF) and LM area (LMA) for BW using data from a series of 7 scans on 24 Suffolk ram lambs born in 2007. Equations were linear, linear + quadratic, allometric (y = alphaBW(beta)), and allometric + BW (ABW; y = alphaBW(beta)e(gammaW)). Goodness of fit was very similar between equations over the range of the data. Resulting adjustment equations were tested using 3 serial scans on winter-born Suffolk (n = 150), Hampshire (n = 36), and Dorset (n = 43) rams and 52 fall-born Dorset rams tested at the Virginia Ram Test in 1999 through 2002. Partial correlations (accounting for the effect of year) between predicted and actual measures ranged from 0.78 to 0.87 for BF and 0.66 to 0.93 for LMA in winter-born rams and from 0.70 to 0.71 for BF and 0.72 to 0.78 for LMA in fall born rams. No significant differences in predictive ability existed between equations for BF or LMA (P > 0.05), and there was no indication that the allometric equation was a better predictor than linear within the range of the data. Adjustment equations were also tested using serial scan data from 37 Suffolk ewe lambs born in the same contemporary group as the rams used to derive the prediction equations but fed for a substantially slower rate of BW gain. Correlations between predicted and actual values of BF and LMA indicated lambs were too young and small at the first scan (77 d, 32.4 kg) to reliably predict carcass measures at typical slaughter weights. For prediction using data from the 2 subsequent scans, at mean ages >96 d and mean BW >39 kg, correlations between predicted and actual values were 0.72 to 0.74 for BF and 0.54 to 0.76 for LMA. Little difference existed between equations for predicting BF. For LMA, the ABW form was a weaker predictor than the others, and the linear equation was slightly superior to allometric. Therefore, it appears the linear and allometric forms are both suitable for use in central ram test and performance-tested farm flocks. PMID- 20023131 TI - Methane emissions, feed intake, and performance of finishing beef cattle offered maize silages harvested at 4 different stages of maturity. AB - This experiment aimed to quantify the methane emissions and intake, digestibility, performance, and carcass characteristics of finishing beef cattle offered maize (Zea mays) silages harvested at 1 of 4 sequential stages of maturity and to relate these values to those obtained from animals offered an ad libitum concentrate-based diet. Sixty continental crossbred steers with a mean initial BW of 531 kg (SD 23.8) were blocked (n = 12 blocks) according to BW and allocated from within block to 1 of 5 dietary treatments in a randomized complete block design: maize silage harvested on September 13 (DM = 277 g/kg), maize silage harvested on September 28 (DM = 315 g/kg), maize silage harvested on October 9 (DM = 339 g/kg), maize silage harvested on October 23 (DM = 333 g/kg), and ad libitum concentrates (ALC). Diets based on maize silage were supplemented with 2.57 kg of concentrate DM daily, and ALC diets were supplemented with 1.27 kg of grass silage DM daily. Silage and total DMI were greater (P = 0.004) with maize silage harvested on September 28 than with any other treatment, which in turn did not differ. Advancing maize maturity at harvest did not affect BW or carcass gain, with the ALC diet exhibiting greater (P = 0.036) rates of carcass gain than any of the maize silage-based treatments. Apparent in vivo digestibility, determined using the AIA indigestible marker technique, was not affected by harvest maturity, with no linear or quadratic trends being identified. Digestibility of DM from the ALC diet was greater (P < 0.001) than with any of the maize silage treatments. Starch digestibility did not differ across maize silage maturities; however, a linear (P = 0.009) decrease in NDF digestibility was observed. Methane emissions, (g/d) measured using the sulfur hexafluoride tracer technique, were not affected by maize silage maturity. Methane emissions relative to DMI tended (P = 0.05) to decline with advancing maize silage maturity, with a similar decline observed when methane was expressed per kilogram of carcass gain. Advancing maize maturity did not result in significant linear or quadratic responses in methane output proportional to GE intake. The ALC diet resulted in less methane output than the maize silage treatments irrespective of the unit of expression. In conclusion, advancing maize harvest maturity did not affect beef cattle performance but reduced methane output relative to DMI and carcass gain. Cattle offered ALC exhibited greater rates of BW gain and less emission of methane compared with cattle offered any of the maize silage treatments. PMID- 20023132 TI - Accounting for additive genetic mutations on litter size in Ripollesa sheep. AB - Little is known about mutational variability in livestock, among which only a few mutations with relatively large effects have been reported. In this manuscript, mutational variability was analyzed in 1,765 litter size records from 404 Ripollesa ewes to characterize the magnitude of this genetic source of variation and check the suitability of including mutational effects in genetic evaluations of this breed. Threshold animal models accounting for additive genetic mutations were preferred to models without mutational contributions, with an average difference in the deviance information criterion of more than 5 units. Moreover, the statistical relevance of the additive genetic mutation term was checked through a Bayes factor approach, which showed that the models with mutational variability were 8.5 to 22.7 times more probable than the others. The mutational heritability (percentage of the phenotypic variance accounted for by mutational variance) was 0.6 or 0.9%, depending on whether genetic dominance effects were accounted for by the analytical model. The inclusion of mutational effects in the genetic model for evaluating litter size in Ripollesa ewes called for some minor modifications in the genetic merit order of the individuals evaluated, which suggested that the continuous uploading of new additive mutations could be taken into account to optimize the selection scheme. This study is the first attempt to estimate mutational variances in a livestock species and thereby contribute to better characterization of the genetic background of productive traits of interest. PMID- 20023133 TI - Effect of dietary supplemental vitamin A concentration on performance, carcass merit, serum metabolites, and lipogenic enzyme activity in yearling beef steers. AB - A randomized complete block design experiment with 360 single-source black yearling steers (average BW = 316.1 +/- 9.1 kg) fed a 91% concentrate (steam flaked corn base) diet was conducted to evaluate the effects of supplemental vitamin A (0, 1,103, 2,205, 4,410, or 8,820 IU/kg of dietary DM) on plasma and liver vitamin A and E concentrations, lipogenic enzyme activity, marbling score, and performance of yearling steers. Final BW (586, 580, 590, 585, and 584 kg for 0, 1,103, 2,205, 4,410, and 8,820 IU of supplemental vitamin A/kg of DM, respectively) did not differ (P = 0.39) among treatments. Feed efficiency, ADG, and daily DMI did not differ (P > 0.10) among treatments within each 28-d period or for the overall experiment. From d 57 to slaughter, average DMI (10.33, 10.28, 10.57, 9.75, and 10.22 kg/steer daily for 0, 1,103, 2,205, 4,410, and 8,820 IU of vitamin A/kg of DM, respectively) was less (P < 0.02) by steers receiving 4,410 IU of supplemental vitamin A/kg of dietary DM than for steers in the other treatments. Furthermore, DMI was greater (P = 0.06) for steers receiving 2,205 IU of supplemental vitamin A/kg of dietary DM than for steers receiving 8,820 IU of supplemental vitamin A/kg of DM. Marbling score, HCW, LM area, and 12th-rib fat thickness did not differ (P > 0.10) among treatments. Similarly, the percentage of carcasses grading >or=USDA Choice (62.6, 52.8, 64.0, 58.4, and 58.4% for 0, 1,103, 2,205, 4,410, and 8,820 IU of vitamin A/kg of DM, respectively), Select, or 0.10) among treatments. Except for d 56 (P = 0.050; r = 0.18 for liver retinol), no correlations (P > 0.10) between marbling score and any plasma or liver tissue retinol or alpha-tocopherol concentrations or vitamin A intake were found, and no differences (P > 0.10) in lipogenic enzyme activity were detected among treatments. Taken together with previous and concurrent research, results of this experiment suggest that vitamin A supplementation at a concentration up to twice the NRC recommendation has little effect on performance, marbling, or lipogenic enzyme activity in adipose tissue samples in yearling feedlot steers, and that 2,205 IU of supplemental vitamin A/kg of DM (20,000 IU/steer daily) or less is adequate to meet the vitamin A requirements of finishing beef cattle. PMID- 20023134 TI - Genetic diversity and population structure of the synthetic Pannon White rabbit revealed by pedigree analyses. AB - Demographic history, current status, and efficiency of the mating strategy were analyzed using the pedigree of Pannon White (PW) rabbits born between 1992 and 2007. Potential accumulation of detrimental effects and loss of genetic diversity were also considered. Calculations and estimates were done most often for rabbits born in 2007, whereas other reference populations (REFPOPXXXX) were specified explicitly. The pedigree contained 4,749 individuals and 580 founders, and its completeness was 82.1% up to 10 and 94.5% up to 5 generations, respectively. Generation intervals through different pathways averaged 1.2 yr. When adjusted to the pedigree completeness, the amount of inbreeding (F(i)) of rabbits was comparable (5.54%) with that of other livestock populations, whereas the 10 (30) founders contributing the most to inbreeding explained a large part of the population inbreeding [i.e., 42.24% (73.18%)]. The ancestral inbreeding coefficient of REFPOP2004 (10.67%) was one-half that of REFPOP2007 (20.66%), showing its strong dependence on pedigree length. Family variance, inbreeding, and realized effective population size were 84.18 (REFPOP2006; this variable could not be calculated for the last year examined), 37.19, and 91.08, respectively. The effective numbers of ancestors, founders, and founder genomes were 48, 26, and 7.33, respectively. Although the circular mating scheme applied was generally effective, the large accumulated reduction in genetic variability indicates the need to revise and improve the current breeding strategy. PMID- 20023135 TI - Effects of distillers dried grains with solubles on amino acid, energy, and fiber digestibility and on hindgut fermentation of dietary fiber in a corn-soybean meal diet fed to growing pigs. AB - The objective of this experiment was to measure the effect of distillers dried grains with solubles (DDGS) on the digestibility of AA, energy, and fiber, on the fermentation of fiber, and on the first appearance of digesta at the end of the ileum, in the cecum, and in the feces of growing pigs fed a corn-soybean meal based diet. Sixteen pigs (initial BW = 38.0 +/- 1.6 kg) were prepared with a T cannula in the distal ileum and a T-cannula in the cecum and allotted to 2 treatments. In period 1, all pigs were fed a corn-soybean meal diet. In periods 2, 3, and 4, pigs were fed the control diet or a diet containing corn, soybean meal, and 30% DDGS. First appearance of digesta at the end of the ileum, in the cecum, and over the entire intestinal tract was measured at the end of period 4. The apparent ileal digestibility (AID) and the apparent total tract digestibility (ATTD) of nutrients were measured, and the concentration of VFA was analyzed in ileal, cecal, and fecal samples. The AID of Lys (74.1%) in the DDGS diet was less (P < 0.05) than in the control diet (78.6%), but the AID of most other AA and GE, NDF, and total dietary fiber (TDF) were not different between the 2 diets. The ATTD of GE (81.0%), NDF (57.2%), TDF (55.5%), and DM (81.7%) were less (P < 0.05) in the DDGS diet than in the control diet (86.0, 69.3, 66.0, and 87.2%, respectively). The concentration of VFA in ileal, cecal, and fecal samples was not different between pigs fed the 2 diets. The pH of ileal and cecal digesta from pigs fed the DDGS diet (6.3 and 5.5) was greater (P < 0.01) than from pigs fed the control diet (5.8 and 5.3). The ATTD of DM, GE, ADF, NDF, and TDF did not change with collection period, but the AID of ADF, NDF, and TDF increased (P < 0.05) from period 2 to period 4. The concentration of all VFA, except isobutyrate, was greater (P < 0.05) in cecal samples from period 4 compared with period 2, and the concentration of all VFA except propionate and isovalerate were greater (P < 0.05) in fecal samples collected in period 4 compared with those collected in period 2. The first appearance of digesta at the end of the ileum, in the cecum, and in the feces was not affected by DDGS. In conclusion, pigs fed the diet containing DDGS had less digestibility of Lys, GE, ADF, NDF, and TDF than pigs fed the control diet. The digestibility of DM and GE was not influenced by collection period, but the concentration of VFA in cecal digesta and feces increased with the length of time pigs received the diets. PMID- 20023136 TI - Developmental programming: the concept, large animal models, and the key role of uteroplacental vascular development. AB - Developmental programming refers to the programming of various bodily systems and processes by a stressor of the maternal system during pregnancy or during the neonatal period. Such stressors include nutritional stress, multiple pregnancy (i.e., increased numbers of fetuses in the gravid uterus), environmental stress (e.g., high environmental temperature, high altitude, prenatal steroid exposure), gynecological immaturity, and maternal or fetal genotype. Programming refers to impaired function of numerous bodily systems or processes, leading to poor growth, altered body composition, metabolic dysfunction, and poor productivity (e.g., poor growth, reproductive dysfunction) of the offspring throughout their lifespan and even across generations. A key component of developmental programming seems to be placental dysfunction, leading to altered fetal growth and development. We discuss various large animal models of developmental programming and how they have and will continue to contribute to our understanding of the mechanisms underlying altered placental function and developmental programming, and, further, how large animal models also will be critical to the identification and application of therapeutic strategies that will alleviate the negative consequences of developmental programming to improve offspring performance in livestock production and human medicine. PMID- 20023137 TI - Mouse AMP-activated protein kinase gamma3 subunit R225Q mutation affecting mouse growth performance when fed a high-energy diet. AB - The Rendement Napole (RN) genotype widely exists in Hampshire pigs. Recently, RN gene was identified as a R200Q mutation in AMP-activated protein kinase (AMPK) gamma3 subunit. The effect of RN genotype on the growth performance of animals and the underlying mechanisms remain controversial. Using transgenic mice carrying an analogous R225Q mutation, the objective of this study was to study the role of RN gene in the growth performance of animals at different energy levels. Wild-type (WT) mice and those with the RN mutation were assigned to 4 groups: 1) WT plus normal diet, 2) RN plus normal diet, 3) WT plus high-energy diet, and 4) RN plus high-energy diet. Mice were weaned at 21 d old and fed the trial diets for 1 mo and then killed for carcass measurements. The pH of postmortem muscle from RN mice was less (P < 0.01) than that from WT mice. No difference in growth performance was observed when mice were fed a normal diet. When fed a high-energy diet, RN mice showed a greater fat accumulation (WT vs. RN, 1.11 vs. 1.63 g for gonadal fat and 1.40 vs. 1.84 g for subcutaneous fat; P < 0.05). Muscle weight was also increased (WT vs. RN, 0.27 vs. 0.30 g for gastrocnemius muscle; P < 0.05). The food consumption was greater in RN compared with WT mice (2.95 vs. 2.49 g; P < 0.05). The AMPK content and its downstream target, acetyl-CoA carboxylase (ACC), content were greater in RN mice (P < 0.05). The phosphorylation of ACC at Ser 79, a site exclusively phosphorylated by AMPK, was increased (P < 0.05), showing greater AMPK activity in RN mouse muscle. No difference in muscle fiber composition and mitochondrial content was observed between WT and RN mice. High fat diet downregulates protein kinase B but upregulates extracellular signal-regulated kinase signaling. In conclusion, the R225Q mutation has no major effect on the growth performance of animals fed a normal diet; a high-energy diet increased fatness in RN mice, likely due to their greater consumption of feed compared with WT mice. PMID- 20023138 TI - Strong associations among rumen endotoxin and acute phase proteins with plasma minerals in lactating cows fed graded amounts of concentrate. AB - The objective of this investigation was to determine associations among rumen endotoxin, plasma serum amyloid A (SAA), and C-reactive protein (CRP) with plasma Ca, Fe, Zn, and Cu in lactating cows challenged with graded amounts of rolled barley grain in the diet (i.e., 0, 15, 30, and 45% of DMI). Correlative relationships among variables were determined by linear and nonlinear regression procedures adjusted for the effects of day, animal, and experimental period. Increasing the amount of grain in the diet was successful in inducing an acute phase response, as assessed by augmentation of rumen endotoxin and plasma CRP and SAA (P < 0.01). The correlative analysis revealed inverse, nonlinear relationships of rumen endotoxin and plasma SAA with circulating Ca. Interestingly, plasma Ca reached the asymptotic plateau at 10.6 mg/dL. The increase in rumen endotoxin was associated with an abrupt decrease in plasma Fe (R(2) = 0.91; P < 0.001). A similar relationship, although at a reduced estimation accuracy (R(2) = 0.21; P < 0.01), was observed between rumen endotoxin and plasma Zn. Augmentation of rumen endotoxin and plasma CRP resulted in a positive, biphasic response of plasma Cu. In conclusion, the increase in rumen endotoxin in response to high-grain diets, and the resulting increases in plasma SAA and CRP, were strongly correlated with fluctuations of plasma minerals. Results suggest that new feeding strategies should be developed to curb the release of endotoxin in the rumen fluid to prevent perturbing minerals in the plasma. PMID- 20023139 TI - Effects of winter growing programs on subsequent feedlot performance, carcass characteristics, body composition, and energy requirements of beef steers. AB - The purpose of this study was to investigate the effects of winter growing program on subsequent finishing performance, carcass merit, and body composition of beef steers. Four steers were slaughtered to determine initial body composition. Remaining steers (n = 256) were blocked by BW and randomly allotted to 1 of 4 treatment groups: 1) ad libitum fed a high-concentrate diet (CF), 2) grazed on wheat pasture (WP), 3) fed a sorghum silage-based diet (SF), or 4) program fed a high-concentrate diet (PF). Steers in the WP, SF, and PF groups were managed to achieve approximately equal rates of BW gain. After the growing phase (112 d), 6 steers were randomly selected from the WP, SF, and PF treatments for determination of body composition. Remaining steers were adapted to a high concentrate diet for finishing and slaughtered at 1.27 cm of 12th-rib fat. Six steers from each treatment were used to determine carcass, offal, and empty body composition. During the growing phase, WP, SF, and PF steers gained 1.15, 1.10, and 1.18 kg/d, respectively, and ME intake did not differ (P = 0.50) among treatments. Program-fed and SF steers had greater (P < 0.05) offal and empty body fat content than WP steers. Gain in offal and empty body fat was greatest (P < 0.05) for PF steers, intermediate for SF steers, and least for WP steers. During the finishing phase (123, 104, 104, 196 d for WP, SF, PF, and CF, respectively) DMI was greater (P < 0.01) for SF steers (10.9 kg/d) than for PF steers (10.1 kg/d); WP steers were intermediate (10.4 kg/d). Daily BW gain was greatest (P < 0.05) for SF steers (2.02 kg/d), intermediate for PF steers (1.85 kg/d), and least for WP and CF steers (1.64 and 1.63 kg/d, respectively). Accretion (kg/d) of carcass and empty body mass was less (P < 0.05) for WP and CF steers compared with PF and SF steers. Calf-fed steers had greater (P < 0.05) fat content of offal than SF and PF steers; WP steers were intermediate. Gain in empty body and carcass energy (Mcal/d) was greater (P < 0.05) for PF steers than CF steers with SF and WP steers being intermediate. At slaughter, SF steers had reduced (P < 0.01) yield grades and greater marbling scores compared with CF and WP steers; PF steers were intermediate. In conclusion, growing programs that increase fat composition of feeder calves did not negatively affect subsequent finishing performance. Finishing steers as calves may reduce retained energy of carcass tissues and increase internal fat during high-grain feeding compared with steers that previously underwent a growing program. PMID- 20023140 TI - Protein digestibility evaluations of meat and fish substrates using laboratory, avian, and ileally cannulated dog assays. AB - Meat and fish serve as important protein sources in the companion animal diet; however, limited protein digestibility data are available for assessing protein digestibility differences among good-quality protein sources. Beef loin, pork loin, chicken breast, pollock fillet, and salmon fillet were evaluated for composition, protein digestibility, and AA bioavailability using the immobilized digestive enzyme assay, cecectomized rooster assay, and ileally cannulated dog assay. Pollock contained the greatest amount of CP, total essential AA (TEAA), and total nonessential AA (TNEAA; DM basis; 96.9, 38.6, and 50.3%, respectively). Salmon contained the next greatest amounts (92.8, 36.4, and 44.6%), followed by chicken (90.3, 36.1, 43.2%). Beef had the least CP content (82.7%), but had slightly greater TEAA and TNEAA concentrations (33.9, 42.0%) compared with pork (86.2, 33.6, 41.3%). Immobilized digestive enzyme assay values were greatest for pollock fillet (0.71) and least for chicken breast (0.52). Beef loin, pork loin, and salmon fillet were similar (0.63, 0.62, and 0.64, respectively). Standardized TEAA and TNEAA digestibility coefficients, evaluated using the cecectomized rooster assay, were greatest (P < 0.05) for pollock fillet (90.4 and 89.8%, respectively) and least (P < 0.05) for chicken breast (86.6 and 85.9%, respectively) and salmon fillet (87.8 and 86.4%, respectively). Dogs assigned to a 5 x 5 Latin square design were fed 5 diets, with each test substrate as the major protein source. No significant differences (P > 0.05) were found in ileal digestibility of protein. Values ranged from 88.9% for chicken to 90.5% for pork loin and pollock fillet. Ileal TEAA and TNEAA coefficients were not different among test substrates, with values between 91.7 and 92.7%, and 88.8 and 90.4%, respectively. Total tract CP apparent digestibility values ranged from 94.4 to 94.8%, with no differences noted among treatments. Despite marked differences in composition and predicted and standardized digestibility values, when the protein sources were added to diets at a concentration of approximately 30% (25% of total energy intake), no differences in test protein substrates were noted in either ileal or total tract nutrient digestibility. PMID- 20023141 TI - Maternal responses to daily maternal porcine somatotropin injections during early mid pregnancy or early-late pregnancy in sows and gilts. AB - Piglet neonatal survival and postnatal growth and efficiency are positively related to birth weight. In gilts, daily maternal porcine ST (pST) injections from d 25 to 100 (term approximately 115 d), but not d 25 to 50, of pregnancy increase progeny birth weight. Daily maternal pST injections from d 25 to 50 increase fetal weight at d 50 in gilts and sows. We therefore hypothesized that daily pST injections from d 25 to 100, but not d 25 to 50, of pregnancy would increase birth weight similarly in both parities. Landrace x Large White gilts and sows were uninjected (controls) or were injected daily with pST (gilts: 2.5 mg/d; sows: 4.0 mg/d, each approximately 15 microg of pST/kg per day) from d 25 to 50 or 100 of pregnancy. Litter size and BW were recorded at birth, midlactation, and weaning. Dams were followed through the subsequent mating and pregnancy. Maternal pST injections from d 25 to 100, but not d 25 to 50, increased mean piglet birth weight by 11.6% in sows (P 0.1) the weaning-remating interval, conception rate, or subsequent litter size. Greater pST-induced birth weight increases in sows than in gilts may mean that underlying metabolic or placental mechanisms for pST action are constrained by maternal competition for nutrients in rapidly growing gilts. PMID- 20023142 TI - Assessment of buffalo semen with the 3-(4,5-dimethylthiazol-2-yl)-2,5 diphenyltetrazolium bromide reduction assay. AB - The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, which measures the reduction of MTT, is commonly used to validate the viability of metabolically active cells. This study was conducted to evaluate and validate the MTT assay to assess the spermatozoal viability of Nili-Ravi buffalo bulls and compare the efficiency of the test with the supravital staining technique (eosin and nigrosin) and the hypoosmotic swelling test. Fresh semen samples from breeding Nili-Ravi buffalo bulls (n = 25) were collected using an artificial vagina. After assessing the quality of the semen for routine variables, the MTT assay was carried out in PBS. Results revealed a correlation (r = 0.979; P < 0.001) between the viability of spermatozoa and the rate of reduction of MTT. The other proportions of same semen samples showed a poor relationship between the eosin and nigrosin test (r = -0.25), the hypoosmotic swelling test (r = -0.12), and motility (r = -0.15). However, the MTT assay was found to be superior compared with other tests because it was able to determine those spermatozoa that were more than 90% viable. In conclusion, the MTT assay is a simple, robust test that can be used to select Nili-Ravi buffalo bulls on the basis of spermatozoa quality. PMID- 20023143 TI - Effects of maternal protein nutrition and subsequent grazing on chicory (Cichorium intybus) on parasitism and performance of lambs. AB - Forty-eight 4- to 5-yr-old Blackface x Bluefaced Leicester (Mule) ewes and their 24-d-old twin lambs were used to assess the effects of maternal protein nutrition and subsequent grazing on chicory (Cichorium intybus) on performance and parasitism. The experiment consisted of 2 grazing periods: safe pasture period and experimental pasture period. During an adaptation period of 66 d, ewes were infected through oral dosing with Teladorsagia circumcincta infective larvae (3 d per wk) and were supplemented with protein (HP) or not (LP) for the last 45 d of this period. At the end of this period, ewes and their lambs were turned out onto a parasitologically safe pasture; all ewes continued to be dosed with parasite (once a week), and HP ewes received protein supplementation for the first 35 d. Ewes and lambs grazed the safe pasture for an additional 43 d after termination of protein supplementation and of oral dosing with parasites. Ewes and their lambs were then moved onto newly established experimental pastures sown with chicory or grass/clover (Lolium perenne/Trifolium repens). During the safe pasture period, HP ewes had decreased fecal egg counts (FEC) compared with LP ewes, whereas HP lambs had temporarily less (P < 0.05) FEC, decreased (P < 0.001) plasma pepsinogen concentrations, and grew faster (P = 0.028) than LP lambs. Lambs grazing chicory had consistently less (P < 0.001) FEC and grew faster (P = 0.013) than lambs grazing grass/clover but had greater (P < 0.001) concentrations of pepsinogen. Pasture larvae counts were decreased (P = 0.07) for the chicory compared with the grass/clover plots. There were no interactions (P > 0.10) between maternal nutrition and grazed forage type on performance or parasitological measurements. Our results suggest that increased maternal protein nutrition and subsequent grazing of chicory independently improve lamb performance and reduce lamb parasitism. PMID- 20023144 TI - Effects of sulfur and monensin concentrations on in vitro dry matter disappearance, hydrogen sulfide production, and volatile fatty acid concentrations in batch culture ruminal fermentations. AB - Effects of monensin (MON) and S on in vitro fermentation and H(2)S production were evaluated in 2 experiments. In Exp. 1, 2 ruminally cannulated steers were adapted (>14 d) to a 75% concentrate diet [steam-flaked corn (SFC)-based], and ruminal fluid was collected approximately 4 h after feeding. Substrate composed (DM basis) of 85.2% SFC, 9% alfalfa hay, 5% cottonseed meal, and 0.8% urea was added with ruminal fluid and buffer to sealed 125-mL serum bottles to allow for gas collection. A Na(2)SO(4) solution was added to yield S equivalent to 0.2, 0.4, and 0.8% of substrate DM, and MON was included at 0, 2, 4, and 6 mg/L of culture volume. Bottle head-space gas was analyzed for H(2)S. No MON (P = 0.29) or MON x S interaction (P = 0.41) effects were detected for H(2)S production. Increasing S linearly increased (P < 0.01) H(2)S production (micromoles/g of fermented DM). The IVDMD (average 70.0%) was not affected by MON (P = 0.93), S (P = 0.18), or the MON x S interaction (P = 0.56). Total VFA concentrations were not affected by MON (P = 0.87), S (P = 0.14), or the MON x S interaction (P = 0.86), but increasing MON linearly decreased (P 21 d) to a 75% concentrate diet (SFC base) that contained 15% (DM basis) wet corn distillers grains plus solubles (WDGS) and MON at 22 mg/kg of DM. In vitro substrate DM was composed of 75.4% SFC, 15% WDGS, 9% alfalfa hay, and 0.6% urea, and S and MON concentrations were the same as in Exp. 1. No effects of MON (P = 0.93) or the MON x S interaction (P = 0.99) were noted for H(2)S production; however, increasing S linearly increased (P < 0.01) H(2)S production. No effects of MON (P = 0.16), S (P = 0.43), or the MON x S interaction (P = 0.10) were noted for IVDMD (average 70.9%). Total VFA concentrations were not affected by MON (P = 0.40), S (P = 0.26), or the MON x S interaction (P = 0.59). As in Exp. 1, increasing MON linearly decreased (P < 0.05) molar proportions of acetate, butyrate, and A:P and linearly increased (P < 0.01) propionate. Increasing S concentration increased in vitro H(2)S production, but S did not affect VFA concentrations and proportions. Monensin did not affect in vitro H(2)S production, and changes in VFA molar proportions were evident with MON regardless of S concentration. PMID- 20023145 TI - The involvement of lipid peroxide-derived aldehydes in aluminum toxicity of tobacco roots. AB - Oxidative injury of the root elongation zone is a primary event in aluminum (Al) toxicity in plants, but the injuring species remain unidentified. We verified the hypothesis that lipid peroxide-derived aldehydes, especially highly electrophilic alpha,beta-unsaturated aldehydes (2-alkenals), participate in Al toxicity. Transgenic tobacco (Nicotiana tabacum) overexpressing Arabidopsis (Arabidopsis thaliana) 2-alkenal reductase (AER-OE plants), wild-type SR1, and an empty vector transformed control line (SR-Vec) were exposed to AlCl(3) on their roots. Compared with the two controls, AER-OE plants suffered less retardation of root elongation under AlCl(3) treatment and showed more rapid regrowth of roots upon Al removal. Under AlCl(3) treatment, the roots of AER-OE plants accumulated Al and H(2)O(2) to the same levels as did the sensitive controls, while they accumulated lower levels of aldehydes and suffered less cell death than SR1 and SR-Vec roots. In SR1 roots, AlCl(3) treatment markedly increased the contents of the highly reactive 2-alkenals acrolein, 4-hydroxy-(E)-2-hexenal, and 4-hydroxy (E)-2-nonenal and other aldehydes such as malondialdehyde and formaldehyde. In AER-OE roots, accumulation of these aldehydes was significantly less. Growth of the roots exposed to 4-hydroxy-(E)-2-nonenal and (E)-2-hexenal were retarded more in SR1 than in AER-OE plants. Thus, the lipid peroxide-derived aldehydes, formed downstream of reactive oxygen species, injured root cells directly. Their suppression by AER provides a new defense mechanism against Al toxicity. PMID- 20023146 TI - The missing link in plant histidine biosynthesis: Arabidopsis myoinositol monophosphatase-like2 encodes a functional histidinol-phosphate phosphatase. AB - Histidine (His) plays a critical role in plant growth and development, both as one of the standard amino acids in proteins, and as a metal-binding ligand. While genes encoding seven of the eight enzymes in the pathway of His biosynthesis have been characterized from a number of plant species, the identity of the enzyme catalyzing the dephosphorylation of histidinol-phosphate to histidinol has remained elusive. Recently, members of a novel family of histidinol-phosphate phosphatase proteins, displaying significant sequence similarity to known myoinositol monophosphatases (IMPs) have been identified from several Actinobacteria. Here we demonstrate that a member of the IMP family from Arabidopsis (Arabidopsis thaliana), myoinositol monophosphatase-like2 (IMPL2; encoded by At4g39120), has histidinol-phosphate phosphatase activity. Heterologous expression of IMPL2, but not the related IMPL1 protein, was sufficient to rescue the His auxotrophy of a Streptomyces coelicolor hisN mutant. Homozygous null impl2 Arabidopsis mutants displayed embryonic lethality, which could be rescued by supplying plants heterozygous for null impl2 alleles with His. In common with the previously characterized HISN genes from Arabidopsis, IMPL2 was expressed in all plant tissues and throughout development, and an IMPL2:green fluorescent protein fusion protein was targeted to the plastid, where His biosynthesis occurs in plants. Our data demonstrate that IMPL2 is the HISN7 gene product, and suggest a lack of genetic redundancy at this metabolic step in Arabidopsis, which is characteristic of the His biosynthetic pathway. PMID- 20023148 TI - Generation of backbone-free, low transgene copy plants by launching T-DNA from the Agrobacterium chromosome. AB - In both applied and basic research, Agrobacterium-mediated transformation is commonly used to introduce genes into plants. We investigated the effect of three Agrobacterium tumefaciens strains and five transferred (T)-DNA origins of replication on transformation frequency, transgene copy number, and the frequency of integration of non-T-DNA portions of the T-DNA-containing vector (backbone) into the genome of Arabidopsis (Arabidopsis thaliana) and maize (Zea mays). Launching T-DNA from the picA locus of the Agrobacterium chromosome increases the frequency of single transgene integration events and almost eliminates the presence of vector backbone sequences in transgenic plants. Along with novel Agrobacterium strains we have developed, our findings are useful for improving the quality of T-DNA integration events. PMID- 20023147 TI - Genomic inventory and transcriptional analysis of Medicago truncatula transporters. AB - Transporters move hydrophilic substrates across hydrophobic biological membranes and play key roles in plant nutrition, metabolism, and signaling and, consequently, in plant growth, development, and responses to the environment. To initiate and support systematic characterization of transporters in the model legume Medicago truncatula, we identified 3,830 transporters and classified 2,673 of these into 113 families and 146 subfamilies. Analysis of gene expression data for 2,611 of these transporters identified 129 that are expressed in an organ specific manner, including 50 that are nodule specific and 36 specific to mycorrhizal roots. Further analysis uncovered 196 transporters that are induced at least 5-fold during nodule development and 44 in roots during arbuscular mycorrhizal symbiosis. Among the nodule- and mycorrhiza-induced transporter genes are many candidates for known transport activities in these beneficial symbioses. The data presented here are a unique resource for the selection and functional characterization of legume transporters. PMID- 20023149 TI - Glucanases and chitinases as causal agents in the protection of Acacia extrafloral nectar from infestation by phytopathogens. AB - Nectars are rich in primary metabolites and attract mutualistic animals, which serve as pollinators or as an indirect defense against herbivores. Their chemical composition makes nectars prone to microbial infestation. As protective strategy, floral nectar of ornamental tobacco (Nicotiana langsdorffii x Nicotiana sanderae) contains "nectarins," proteins producing reactive oxygen species such as hydrogen peroxide. By contrast, pathogenesis-related (PR) proteins were detected in Acacia extrafloral nectar (EFN), which is secreted in the context of defensive ant-plant mutualisms. We investigated whether these PR proteins protect EFN from phytopathogens. Five sympatric species (Acacia cornigera, A. hindsii, A. collinsii, A. farnesiana, and Prosopis juliflora) were compared that differ in their ant-plant mutualism. EFN of myrmecophytes, which are obligate ant-plants that secrete EFN constitutively to nourish specialized ant inhabitants, significantly inhibited the growth of four out of six tested phytopathogenic microorganisms. By contrast, EFN of nonmyrmecophytes, which is secreted only transiently in response to herbivory, did not exhibit a detectable inhibitory activity. Combining two-dimensional sodium dodecyl sulfate-polyacrylamide gel electrophoresis with nanoflow liquid chromatography-tandem mass spectrometry analysis confirmed that PR proteins represented over 90% of all proteins in myrmecophyte EFN. The inhibition of microbial growth was exerted by the protein fraction, but not the small metabolites of this EFN, and disappeared when nectar was heated. In-gel assays demonstrated the activity of acidic and basic chitinases in all EFNs, whereas glucanases were detected only in EFN of myrmecophytes. Our results demonstrate that PR proteins causally underlie the protection of Acacia EFN from microorganisms and that acidic and basic glucanases likely represent the most important prerequisite in this defensive function. PMID- 20023150 TI - AtMetExpress development: a phytochemical atlas of Arabidopsis development. AB - Plants possess many metabolic genes for the production of a wide variety of phytochemicals in a tissue-specific manner. However, the metabolic systems behind the diversity and tissue-dependent regulation still remain unknown due to incomplete characterization of phytochemicals produced in a single plant species. Thus, having a metabolome dataset in addition to the genome and transcriptome information resources would enrich our knowledge of plant secondary metabolism. Here we analyzed phytochemical accumulation during development of the model plant Arabidopsis (Arabidopsis thaliana) using liquid chromatography-mass spectrometry in samples covering many growth stages and organs. We also obtained tandem mass spectrometry spectral tags of many metabolites as a resource for elucidation of metabolite structure. These are part of the AtMetExpress metabolite accumulation atlas. Based on the dataset, we detected 1,589 metabolite signals from which the structures of 167 metabolites were elucidated. The integrated analyses with transcriptome data demonstrated that Arabidopsis produces various phytochemicals in a highly tissue-specific manner, which often accompanies the expression of key biosynthesis-related genes. We also found that a set of biosynthesis-related genes is coordinately expressed among the tissues. These data suggested that the simple mode of regulation, transcript to metabolite, is an origin of the dynamics and diversity of plant secondary metabolism. PMID- 20023151 TI - Tryptophan-derived metabolites are required for antifungal defense in the Arabidopsis mlo2 mutant. AB - Arabidopsis (Arabidopsis thaliana) genes MILDEW RESISTANCE LOCUS O2 (MLO2), MLO6, and MLO12 exhibit unequal genetic redundancy with respect to the modulation of defense responses against powdery mildew fungi and the control of developmental phenotypes such as premature leaf decay. We show that early chlorosis and necrosis of rosette leaves in mlo2 mlo6 mlo12 mutants reflects an authentic but untimely leaf senescence program. Comparative transcriptional profiling revealed that transcripts of several genes encoding tryptophan biosynthetic and metabolic enzymes hyperaccumulate during vegetative development in the mlo2 mlo6 mlo12 mutant. Elevated expression levels of these genes correlate with altered steady state levels of several indolic metabolites, including the phytoalexin camalexin and indolic glucosinolates, during development in the mlo2 single mutant and the mlo2 mlo6 mlo12 triple mutant. Results of genetic epistasis analysis suggest a decisive role for indolic metabolites in mlo2-conditioned antifungal defense against both biotrophic powdery mildews and a camalexin-sensitive strain of the necrotrophic fungus Botrytis cinerea. The wound- and pathogen-responsive callose synthase POWDERY MILDEW RESISTANCE4/GLUCAN SYNTHASE-LIKE5 was found to be responsible for the spontaneous callose deposits in mlo2 mutant plants but dispensable for mlo2-conditioned penetration resistance. Our data strengthen the notion that powdery mildew resistance of mlo2 genotypes is based on the same defense execution machinery as innate antifungal immune responses that restrict the invasion of nonadapted fungal pathogens. PMID- 20023154 TI - MHCalpha and cardiogenesis. PMID- 20023155 TI - Dissecting the regulatory switches of development: lessons from enhancer evolution in Drosophila. AB - Cis-regulatory modules are non-protein-coding regions of DNA essential for the control of gene expression. One class of regulatory modules is embryonic enhancers, which drive gene expression during development as a result of transcription factor protein binding at the enhancer sequences. Recent comparative studies have begun to investigate the evolution of the sequence architecture within enhancers. These analyses are illuminating the way that developmental biologists think about enhancers by revealing their molecular mechanism of function. PMID- 20023156 TI - Promoting developmental transcription. AB - Animal growth and development depend on the precise control of gene expression at the level of transcription. A central role in the regulation of developmental transcription is attributed to transcription factors that bind DNA enhancer elements, which are often located far from gene transcription start sites. Here, we review recent studies that have uncovered significant regulatory functions in developmental transcription for the TFIID basal transcription factors and for the DNA core promoter elements that are located close to transcription start sites. PMID- 20023157 TI - Hematopoietic stem cells in Drosophila. AB - The Drosophila lymph gland, the source of adult hemocytes, is established by mid embryogenesis. During larval stages, a pool of pluripotent hemocyte precursors differentiate into hemocytes that are released into circulation upon metamorphosis or in response to immune challenge. This process is controlled by the posterior signaling center (PSC), which is reminiscent of the vertebrate hematopoietic stem cell niche. Using lineage analysis, we identified bona fide hematopoietic stem cells (HSCs) in the lymph glands of embryos and young larvae, which give rise to a hematopoietic lineage. These lymph glands also contain pluripotent precursor cells that undergo a limited number of mitotic divisions and differentiate. We further find that the conserved factor Zfrp8/PDCD2 is essential for the maintenance of the HSCs, but dispensable for their daughter cells, the pluripotent precursors. Zfrp8/PDCD2 is likely to have similar functions in hematopoietic stem cell maintenance in vertebrates. PMID- 20023158 TI - FGF-receptor signalling controls neural cell diversity in the zebrafish hindbrain by regulating olig2 and sox9. AB - The mechanisms underlying the generation of neural cell diversity are the subject of intense investigation, which has highlighted the involvement of different signalling molecules including Shh, BMP and Wnt. By contrast, relatively little is known about FGF in this process. In this report we identify an FGF-receptor dependent pathway in zebrafish hindbrain neural progenitors that give rise to somatic motoneurons, oligodendrocyte progenitors and differentiating astroglia. Using a combination of chemical and genetic approaches to conditionally inactivate FGF-receptor signalling, we investigate the role of this pathway. We show that FGF-receptor signalling is not essential for the survival or maintenance of hindbrain neural progenitors but controls their fate by coordinately regulating key transcription factors. First, by cooperating with Shh, FGF-receptor signalling controls the expression of olig2, a patterning gene essential for the specification of somatic motoneurons and oligodendrocytes. Second, FGF-receptor signalling controls the development of both oligodendrocyte progenitors and astroglia through the regulation of sox9, a gliogenic transcription factor the function of which we show to be conserved in the zebrafish hindbrain. Overall, for the first time in vivo, our results reveal a mechanism of FGF in the control of neural cell diversity. PMID- 20023159 TI - Lineage-specific effects of Notch/Numb signaling in post-embryonic development of the Drosophila brain. AB - Numb can antagonize Notch signaling to diversify the fates of sister cells. We report here that paired sister cells acquire different fates in all three Drosophila neuronal lineages that make diverse types of antennal lobe projection neurons (PNs). Only one in each pair of postmitotic neurons survives into the adult stage in both anterodorsal (ad) and ventral (v) PN lineages. Notably, Notch signaling specifies the PN fate in the vPN lineage but promotes programmed cell death in the missing siblings in the adPN lineage. In addition, Notch/Numb mediated binary sibling fates underlie the production of PNs and local interneurons from common precursors in the lAL lineage. Furthermore, Numb is needed in the lateral but not adPN or vPN lineages to prevent the appearance of ectopic neuroblasts and to ensure proper self-renewal of neural progenitors. These lineage-specific outputs of Notch/Numb signaling show that a universal mechanism of binary fate decision can be utilized to govern diverse neural sibling differentiations. PMID- 20023160 TI - Role of Notch signaling in establishing the hemilineages of secondary neurons in Drosophila melanogaster. AB - The secondary neurons generated in the thoracic central nervous system of Drosophila arise from a hemisegmental set of 25 neuronal stem cells, the neuroblasts (NBs). Each NB undergoes repeated asymmetric divisions to produce a series of smaller ganglion mother cells (GMCs), which typically divide once to form two daughter neurons. We find that the two daughters of the GMC consistently have distinct fates. Using both loss-of-function and gain-of-function approaches, we examined the role of Notch signaling in establishing neuronal fates within all of the thoracic secondary lineages. In all cases, the 'A' (Notch(ON)) sibling assumes one fate and the 'B' (Notch(OFF)) sibling assumes another, and this relationship holds throughout the neurogenic period, resulting in two major neuronal classes: the A and B hemilineages. Apparent monotypic lineages typically result from the death of one sibling throughout the lineage, resulting in a single, surviving hemilineage. Projection neurons are predominantly from the B hemilineages, whereas local interneurons are typically from A hemilineages. Although sibling fate is dependent on Notch signaling, it is not necessarily dependent on numb, a gene classically involved in biasing Notch activation. When Numb was removed at the start of larval neurogenesis, both A and B hemilineages were still generated, but by the start of the third larval instar, the removal of Numb resulted in all neurons assuming the A fate. The need for Numb to direct Notch signaling correlated with a decrease in NB cell cycle time and may be a means for coping with multiple sibling pairs simultaneously undergoing fate decisions. PMID- 20023161 TI - Auxin modulates the transition from the mitotic cycle to the endocycle in Arabidopsis. AB - Amplification of genomic DNA by endoreduplication often marks the initiation of cell differentiation in animals and plants. The transition from mitotic cycles to endocycles should be developmentally programmed but how this process is regulated remains largely unknown. We show that the plant growth regulator auxin modulates the switch from mitotic cycles to endocycles in Arabidopsis; high levels of TIR1 AUX/IAA-ARF-dependent auxin signalling are required to repress endocycles, thus maintaining cells in mitotic cycles. By contrast, lower levels of TIR1-AUX/IAA ARF-dependent auxin signalling trigger an exit from mitotic cycles and an entry into endocycles. Our data further demonstrate that this auxin-mediated modulation of the mitotic-to-endocycle switch is tightly coupled with the developmental transition from cell proliferation to cell differentiation in the Arabidopsis root meristem. The transient reduction of auxin signalling by an auxin antagonist PEO-IAA rapidly downregulates the expression of several core cell cycle genes, and we show that overexpressing one of the genes, CYCLIN A2;3 (CYCA2;3), partially suppresses an early initiation of cell differentiation induced by PEO IAA. Taken together, these results suggest that auxin-mediated mitotic-to endocycle transition might be part of the developmental programmes that balance cell proliferation and cell differentiation in the Arabidopsis root meristem. PMID- 20023162 TI - DNA LIGASE I exerts a maternal effect on seed development in Arabidopsis thaliana. AB - Maternal effects are defined by mutations that affect the next generation when they are maternally inherited. To date, most indepth studies of maternal effects in plants have attributed their origin to genomic imprinting that restricts expression to the maternal allele. The DNA glycosylase DEMETER (DME) removes methylated cytosine residues, causing transcriptional activation of the maternal allele of imprinted genes. In this study, we show that loss-of-function of the major DNA LIGASE I (AtLIG1) in Arabidopsis thaliana causes maternal effects in the endosperm, which is the seed tissue that nurtures embryo development. AtLIG1 expression is not imprinted and has a limited impact on imprinted gene expression. Genetic interaction analyses further indicate that AtLIG1 acts downstream of DME. The removal of methylated cytosine residues by DME involves the creation of DNA single-strand breaks and our results suggest that AtLIG1 repairs these breaks. PMID- 20023163 TI - Dynamics of Delta/Notch signaling on endomesoderm segregation in the sea urchin embryo. AB - Endomesoderm is the common progenitor of endoderm and mesoderm early in the development of many animals. In the sea urchin embryo, the Delta/Notch pathway is necessary for the diversification of this tissue, as are two early transcription factors, Gcm and FoxA, which are expressed in mesoderm and endoderm, respectively. Here, we provide a detailed lineage analysis of the cleavages leading to endomesoderm segregation, and examine the expression patterns and the regulatory relationships of three known regulators of this cell fate dichotomy in the context of the lineages. We observed that endomesoderm segregation first occurs at hatched blastula stage. Prior to this stage, Gcm and FoxA are co expressed in the same cells, whereas at hatching these genes are detected in two distinct cell populations. Gcm remains expressed in the most vegetal endomesoderm descendant cells, while FoxA is downregulated in those cells and activated in the above neighboring cells. Initially, Delta is expressed exclusively in the micromeres, where it is necessary for the most vegetal endomesoderm cell descendants to express Gcm and become mesoderm. Our experiments show a requirement for a continuous Delta input for more than two cleavages (or about 2.5 hours) before Gcm expression continues in those cells independently of further Delta input. Thus, this study provides new insights into the timing mechanisms and the molecular dynamics of endomesoderm segregation during sea urchin embryogenesis and into the mode of action of the Delta/Notch pathway in mediating mesoderm fate. PMID- 20023164 TI - C. elegans STRADalpha and SAD cooperatively regulate neuronal polarity and synaptic organization. AB - Neurons are polarized cells with morphologically and functionally distinct axons and dendrites. The SAD kinases are crucial for establishing the axon-dendrite identity across species. Previous studies suggest that a tumour suppressor kinase, LKB1, in the presence of a pseudokinase, STRADalpha, initiates axonal differentiation and growth through activating the SAD kinases in vertebrate neurons. STRADalpha was implicated in the localization, stabilization and activation of LKB1 in various cell culture studies. Its in vivo functions, however, have not been examined. In our present study, we analyzed the neuronal phenotypes of the first loss-of-function mutants for STRADalpha and examined their genetic interactions with LKB1 and SAD in C. elegans. Unexpectedly, only the C. elegans STRADalpha, STRD-1, functions exclusively through the SAD kinase, SAD-1, to regulate neuronal polarity and synaptic organization. Moreover, STRD-1 tightly associates with SAD-1 to coordinate its synaptic localizations. By contrast, the C. elegans LKB1, PAR-4, also functions in an additional genetic pathway independently of SAD-1 and STRD-1 to regulate neuronal polarity. We propose that STRD-1 establishes neuronal polarity and organizes synaptic proteins in a complex with the SAD-1 kinase. Our findings suggest that instead of a single, linear genetic pathway, STRADalpha and LKB1 regulate neuronal development through multiple effectors that are shared in some cellular contexts but distinct in others. PMID- 20023165 TI - Control of cell proliferation in Arabidopsis thaliana by microRNA miR396. AB - Cell proliferation is an important determinant of plant form, but little is known about how developmental programs control cell division. Here, we describe the role of microRNA miR396 in the coordination of cell proliferation in Arabidopsis leaves. In leaf primordia, miR396 is expressed at low levels that steadily increase during organ development. We found that miR396 antagonizes the expression pattern of its targets, the GROWTH-REGULATING FACTOR (GRF) transcription factors. miR396 accumulates preferentially in the distal part of young developing leaves, restricting the expression of GRF2 to the proximal part of the organ. This, in turn, coincides with the activity of the cell proliferation marker CYCLINB1;1. We show that miR396 attenuates cell proliferation in developing leaves, through the repression of GRF activity and a decrease in the expression of cell cycle genes. We observed that the balance between miR396 and the GRFs controls the final number of cells in leaves. Furthermore, overexpression of miR396 in a mutant lacking GRF-INTERACTING FACTOR 1 severely compromises the shoot meristem. We found that miR396 is expressed at low levels throughout the meristem, overlapping with the expression of its target, GRF2. In addition, we show that miR396 can regulate cell proliferation and the size of the meristem. Arabidopsis plants with an increased activity of the transcription factor TCP4, which reduces cell proliferation in leaves, have higher miR396 and lower GRF levels. These results implicate miR396 as a significant module in the regulation of cell proliferation in plants. PMID- 20023166 TI - The MED12-MED13 module of Mediator regulates the timing of embryo patterning in Arabidopsis. AB - The Arabidopsis embryo becomes patterned into central and peripheral domains during the first few days after fertilization. A screen for mutants that affect this process identified two genes, GRAND CENTRAL (GCT)and CENTER CITY (CCT). Mutations in GCT and CCT delay the specification of central and peripheral identity and the globular-to-heart transition, but have little or no effect on the initial growth rate of the embryo. Mutant embryos eventually recover and undergo relatively normal patterning, albeit at an inappropriate size. GCT and CCT were identified as the Arabidopsis orthologs of MED13 and MED12 - evolutionarily conserved proteins that act in association with the Mediator complex to negatively regulate transcription. The predicted function of these proteins combined with the effect of gct and cct on embryo development suggests that MED13 and MED12 regulate pattern formation during Arabidopsis embryogenesis by transiently repressing a transcriptional program that interferes with this process. Their mutant phenotype reveals the existence of a previously unknown temporal regulatory mechanism in plant embryogenesis. PMID- 20023167 TI - The Drosophila nuclear receptors DHR3 and betaFTZ-F1 control overlapping developmental responses in late embryos. AB - Studies of the onset of metamorphosis have identified an ecdysone-triggered transcriptional cascade that consists of the sequential expression of the transcription-factor-encoding genes DHR3, betaFTZ-F1, E74A and E75A. Although the regulatory interactions between these genes have been well characterized by genetic and molecular studies over the past 20 years, their developmental functions have remained more poorly understood. In addition, a transcriptional sequence similar to that observed in prepupae is repeated before each developmental transition in the life cycle, including mid-embryogenesis and the larval molts. Whether the regulatory interactions between DHR3, betaFTZ-F1, E74A and E75A at these earlier stages are similar to those defined at the onset of metamorphosis, however, is unknown. In this study, we turn to embryonic development to address these two issues. We show that mid-embryonic expression of DHR3 and betaFTZ-F1 is part of a 20-hydroxyecdysone (20E)-triggered transcriptional cascade similar to that seen in mid-prepupae, directing maximal expression of E74A and E75A during late embryogenesis. In addition, DHR3 and betaFTZ-F1 exert overlapping developmental functions at the end of embryogenesis. Both genes are required for tracheal air filling, whereas DHR3 is required for ventral nerve cord condensation and betaFTZ-F1 is required for proper maturation of the cuticular denticles. Rescue experiments support these observations, indicating that DHR3 has essential functions independent from those of betaFTZ F1. DHR3 and betaFTZ-F1 also contribute to overlapping transcriptional responses during embryogenesis. Taken together, these studies define the lethal phenotypes of DHR3 and betaFTZ-F1 mutants, and provide evidence for functional bifurcation in the 20E-responsive transcriptional cascade. PMID- 20023168 TI - Evidence for a role of vertebrate Disp1 in long-range Shh signaling. AB - Dispatched 1 (Disp1) encodes a twelve transmembrane domain protein that is required for long-range sonic hedgehog (Shh) signaling. Inhibition of Disp1 function, both by RNAi or dominant-negative constructs, prevents secretion and results in the accumulation of Shh in source cells. Measuring the Shh response in neuralized embryoid bodies (EBs) derived from embryonic stem (ES) cells, with or without Disp1 function, demonstrates an additional role for Disp1 in cells transporting Shh. Co-cultures with Shh-expressing cells revealed a significant reduction in the range of the contact-dependent Shh response in Disp1(-/-) neuralized EBs. These observations support a dual role for Disp1, not only in the secretion of Shh from the source cells, but also in the subsequent transport of Shh through tissue. PMID- 20023169 TI - Interaction between Drosophila bZIP proteins Atf3 and Jun prevents replacement of epithelial cells during metamorphosis. AB - Epithelial sheet spreading and fusion underlie important developmental processes. Well-characterized examples of such epithelial morphogenetic events have been provided by studies in Drosophila, and include embryonic dorsal closure, formation of the adult thorax and wound healing. All of these processes require the basic region-leucine zipper (bZIP) transcription factors Jun and Fos. Much less is known about morphogenesis of the fly abdomen, which involves replacement of larval epidermal cells (LECs) with adult histoblasts that divide, migrate and finally fuse to form the adult epidermis during metamorphosis. Here, we implicate Drosophila Activating transcription factor 3 (Atf3), the single ortholog of human ATF3 and JDP2 bZIP proteins, in abdominal morphogenesis. During the process of the epithelial cell replacement, transcription of the atf3 gene declines. When this downregulation is experimentally prevented, the affected LECs accumulate cell-adhesion proteins and their extrusion and replacement with histoblasts are blocked. The abnormally adhering LECs consequently obstruct the closure of the adult abdominal epithelium. This closure defect can be either mimicked and further enhanced by knockdown of the small GTPase Rho1 or, conversely, alleviated by stimulating ecdysone steroid hormone signaling. Both Rho and ecdysone pathways have been previously identified as effectors of the LEC replacement. To elicit the gain-of-function effect, Atf3 specifically requires its binding partner Jun. Our data thus identify Atf3 as a new functional partner of Drosophila Jun during development. PMID- 20023170 TI - G protein-coupled receptor 48 upregulates estrogen receptor alpha expression via cAMP/PKA signaling in the male reproductive tract. AB - The epididymis and efferent ducts play major roles in sperm maturation, transport, concentration and storage by reabsorbing water, ions and proteins produced from seminiferous tubules. Gpr48-null male mice demonstrate reproductive tract defects and infertility. In the present study, we found that estrogen receptor alpha (ERalpha) was dramatically reduced in the epididymis and efferent ducts in Gpr48-null male mice. We further revealed that ERalpha could be upregulated by Gpr48 activation via the cAMP/PKA signaling pathway. Moreover, we identified a cAMP responsive element (Cre) motif located at -1307 to -1300 bp in the ERalpha promoter that is able to interact with Cre binding protein (Creb). In conclusion, Gpr48 participates in the development of the male epididymis and efferent ducts through regulation of ERalpha expression via the cAMP/PKA signaling pathway. PMID- 20023171 TI - A novel regulatory mechanism for Fgf18 signaling involving cysteine-rich FGF receptor (Cfr) and delta-like protein (Dlk). AB - Fibroblast growth factors (FGFs) transduce signals through FGF receptors (FGFRs) and have pleiotropic functions. Besides signal-transducing FGFRs, cysteine-rich FGF receptor (Cfr; Glg1) is also known to bind some FGFs, although its physiological functions remain unknown. In this study, we generated Cfr-deficient mice and found that some of them die perinatally, and show growth retardation, tail malformation and cleft palate. These phenotypes are strikingly similar to those of Fgf18-deficient mice, and we revealed interaction between Cfr and Fgf18 both genetically and physically, suggesting functional cooperation. Consistently, introduction of Cfr facilitated Fgf18-dependent proliferation of Ba/F3 cells expressing Fgfr3c. In addition, we uncovered binding between Cfr and delta-like protein (Dlk), and noticed that Cfr-deficient mice are also similar to Dlk transgenic mice, indicating that Cfr and Dlk function in opposite ways. Interestingly, we also found that Dlk interrupts the binding between Cfr and Fgf18. Thus, the Fgf18 signaling pathway seems to be finely tuned by Cfr and Dlk for skeletal development. This study reveals a novel regulatory mechanism for Fgf18 signaling involving Cfr and Dlk. PMID- 20023172 TI - Distinguishing direct from indirect roles for bicoid mRNA localization factors. AB - Localization of bicoid mRNA to the anterior of the Drosophila oocyte is essential for patterning the anteroposterior body axis in the early embryo. bicoid mRNA localizes in a complex multistep process involving transacting factors, molecular motors and cytoskeletal components that remodel extensively during the lifetime of the mRNA. Genetic requirements for several localization factors, including Swallow and Staufen, are well established, but the precise roles of these factors and their relationship to bicoid mRNA transport particles remains unresolved. Here we use live cell imaging, super-resolution microscopy in fixed cells and immunoelectron microscopy on ultrathin frozen sections to study the distribution of Swallow, Staufen, actin and dynein relative to bicoid mRNA during late oogenesis. We show that Swallow and bicoid mRNA are transported independently and are not colocalized at their final destination. Furthermore, Swallow is not required for bicoid transport. Instead, Swallow localizes to the oocyte plasma membrane, in close proximity to actin filaments, and we present evidence that Swallow functions during the late phase of bicoid localization by regulating the actin cytoskeleton. In contrast, Staufen, dynein and bicoid mRNA form nonmembranous, electron dense particles at the oocyte anterior. Our results exclude a role for Swallow in linking bicoid mRNA to the dynein motor. Instead we propose a model for bicoid mRNA localization in which Swallow is transported independently by dynein and contributes indirectly to bicoid mRNA localization by organizing the cytoskeleton, whereas Staufen plays a direct role in dynein dependent bicoid mRNA transport. PMID- 20023173 TI - Role of protein tyrosine phosphatase SHP2 in barrier function of pulmonary endothelium. AB - Pulmonary edema is mediated in part by disruption of interendothelial cell contacts. Protein tyrosine phosphatases (PTP) have been shown to affect both cell extracellular matrix and cell-cell junctions. The SH2 domain-containing nonreceptor PTP, SHP2, is involved in intercellular signaling through direct interaction with adherens junction proteins. In this study, we examined the role of SHP2 in pulmonary endothelial barrier function. Inhibition of SHP2 promoted edema formation in rat lungs and increased monolayer permeability in cultured lung endothelial cells. In addition, pulmonary endothelial cells demonstrated a decreased level of p190RhoGAP activity following inhibition of SHP2, events that were accompanied by a concomitant increase in RhoA activity. Furthermore, immunofluorescence microscopy confirmed enhanced actin stress fiber formation and diminished interendothelial staining of adherens junction complex-associated proteins upon SHP2 inhibition. Finally, immunoprecipitation and immunoblot analyses demonstrated increased tyrosine phosphorylation of VE-cadherin, beta catenin, and p190RhoGAP proteins, as well as decreased association between p120 catenin and VE-cadherin proteins. Our findings suggest that SHP2 supports basal pulmonary endothelial barrier function by coordinating the tyrosine phosphorylation profile of VE-cadherin, beta-catenin, and p190RhoGAP and the activity of RhoA, signaling molecules important in adherens junction complex integrity. PMID- 20023174 TI - Muscarinic receptor-mediated bronchoconstriction is coupled to caveolae in murine airways. AB - Cholinergic bronchoconstriction is mediated by M(2) and M(3) muscarinic receptors (MR). In heart and urinary bladder, MR are linked to caveolin-1 or -3, the structural proteins of caveolae. Caveolae are cholesterol-rich, omega-shaped invaginations of the plasma membrane. They provide a scaffold for multiple G protein receptors and membrane-bound enzymes, thereby orchestrating signaling into the cell interior. Hence, we hypothesized that airway MR signaling pathways are coupled to caveolae as well. To address this issue, we determined the distribution of caveolin isoforms and MR subtype M2R in murine and human airways and investigated protein-protein associations by fluorescence resonance energy transfer (FRET)-confocal laser scanning microscopy (CLSM) analysis in immunolabeled murine tissue sections. Bronchoconstrictor responses of murine bronchi were recorded in lung-slice preparations before and after caveolae disruption by methyl-beta-cyclodextrin, with efficiency of this treatment being validated by electron microscopy. KCl-induced bronchoconstriction was unaffected after treatment, demonstrating functional integrity of the smooth muscle. Caveolae disruption decreased muscarine-induced bronchoconstriction in wild-type and abolished it in M2R(-/-) and M3R(-/-) mice. Thus M2R and M3R signaling pathways require intact caveolae. Furthermore, we identified a presumed skeletal and cardiac myocyte-specific caveolin isoform, caveolin-3, in human and murine bronchial smooth muscle and found it to be associated with M2R in situ. In contrast, M2R was not associated with caveolin-1, despite an in situ association of caveolin-1 and caveolin-3 that was detected. Here, we demonstrated that M2R- and M3R-mediated bronchoconstriction is caveolae-dependent. Since caveolin-3 is directly associated with M2R, we suggest caveolin-3 as novel regulator of M2R mediated signaling. PMID- 20023175 TI - Functional importance of the NH2-terminal insertion sequence of lung surfactant protein B. AB - Lung surfactant protein B (SP-B) is required for proper surface activity of pulmonary surfactant. In model lung surfactant lipid systems composed of saturated and unsaturated lipids, the unsaturated lipids are removed from the film at high compression. It is thought that SP-B helps anchor these lipids closely to the monolayer in three-dimensional cylindrical structures termed "nanosilos" seen by atomic force microscopy imaging of deposited monolayers at high surface pressures. Here we explore the role of the SP-B NH(2) terminus in the formation and stability of these cylindrical structures, specifically the distribution of lipid stack height, width, and density with four SP-B truncation peptides: SP-B 1-25, SP-B 9-25, SP-B 11-25, and SP-B 1-25Nflex (prolines 2 and 4 substituted with alanine). The first nine amino acids, termed the insertion sequence and the interface seeking tryptophan residue 9, are shown to stabilize the formation of nanosilos while an increase in the insertion sequence flexibility (SP-B 1-25Nflex) may improve peptide functionality. This provides a functional understanding of the insertion sequence beyond anchoring the protein to the two-dimensional membrane lining the lung, as it also stabilizes formation of nanosilos, creating reversible repositories for fluid lipids at high compression. In lavaged, surfactant-deficient rats, instillation of a mixture of SP-B 1-25 (as a monomer or dimer) and synthetic lung lavage lipids quickly improved oxygenation and dynamic compliance, whereas SP-B 11-25 surfactants showed oxygenation and dynamic compliance values similar to that of lipids alone, demonstrating a positive correlation between formation of stable, but reversible, nanosilos and in vivo efficacy. PMID- 20023177 TI - Gene expression profile in flow-associated pulmonary arterial hypertension with neointimal lesions. AB - Pulmonary arterial hypertension (PAH) is a pulmonary angioproliferative disease with high morbidity and mortality, characterized by a typical pattern of pulmonary vascular remodeling including neointimal lesions. In congenital heart disease, increased pulmonary blood flow has appeared to be a key mediator in the development of these characteristic lesions, but the molecular mechanisms underlying the pulmonary vascular lesions are largely unknown. We employed a rat model of flow-associated PAH, which induced specific pulmonary neointimal lesions. We identified gene expression profiles in rats specifically related to the addition of increased pulmonary blood flow to monocrotaline and the associated occurrence of neointimal lesions. Increased pulmonary blood flow induced the expression of the transcription factors activating transcription factor-3 (ATF3) and early growth response factor-1 (EGR-1), for which presence was confirmed in neointimal lesions. Monocrotaline alone induced increased numbers of activated mast cells and their products. We further identified molecular pathways that may be involved in treatment with the prostacyclin analog iloprost, a vasoactive compound with clinically beneficial effects in patients with PAH, which were similar to pathways described in samples from patient studies. These pathways, associated with the development of angioproliferative lesions as well as with the response to therapy in PAH, may provide new therapeutic targets. PMID- 20023176 TI - Transforming growth factor-beta modulates the expression of nitric oxide signaling enzymes in the injured developing lung and in vascular smooth muscle cells. AB - Nitric oxide signaling has an important role in regulating pulmonary development and function. Expression of soluble guanylate cyclase (sGC) and cGMP-dependent protein kinase I (PKGI), both critical mediators of nitric oxide (NO) signaling, is diminished in the injured newborn lung through unknown mechanisms. Recent studies suggest that excessive transforming growth factor-beta (TGF-beta) activity inhibits injured newborn lung development. To explore mechanisms that regulate pulmonary NO signaling, we tested whether TGF-beta decreases sGC and PKGI expression in the injured developing lung and pulmonary vascular smooth muscle cells (SMC). We found that chronic oxygen-induced lung injury decreased pulmonary sGCalpha(1) and PKGI immunoreactivity in mouse pups and that exposure to a TGF-beta-neutralizing antibody prevented this reduction of sGC and PKGI protein expression. In addition, TGF-beta(1) decreased expression of NO signaling enzymes in freshly isolated pulmonary microvascular SMC/myofibroblasts, suggesting that TGF-beta has a direct role in modulating NO signaling in the pup lung. Moreover, TGF-beta(1) decreased sGC and PKGI expression in pulmonary artery and aortic SMC from adult rats and mice, suggesting a general role for TGF-beta in modulating NO signaling in vascular SMC. Although other cytokines decrease sGC mRNA stability, TGF-beta did not modulate sGCalpha(1) or PKGIbeta mRNA turnover in vascular SMC. These studies indicate for the first time that TGF-beta decreases NO signaling enzyme expression in the injured developing lung and pulmonary vascular SMC. Moreover, they suggest that TGF-beta-neutralizing molecules might counteract the effects of injury on NO signaling in the newborn lung. PMID- 20023178 TI - Coping styles and strategies: a comparison of adolescent students with and without learning disabilities. AB - In this study, the authors compared the results of a coping measure completed by 98 seventh through ninth grade students who were assessed as having learning disabilities with published means from the general Australian student population. The Adolescent Coping Scale was the measure used. The results suggested higher use by students aged 12 to 13 years who had learning disabilities of an overall nonproductive coping style and in particular of the nonproductive strategies of ignoring the problem and not coping. Although there was no difference in overall productive or nonproductive coping style for 14- to 15-year-olds, the students in this age group who had learning disabilities reported higher use of the strategies of not coping and ignoring the problem. These findings are discussed in relation to a need for interventions that give students who have learning disabilities strategies that address the risk of a passive coping style. PMID- 20023179 TI - Interesting insights from a very simple (unicellular) source. PMID- 20023180 TI - The Hippocratic paradigm in medicine: origins of the clinical encounter. PMID- 20023181 TI - Plasma exchange for heparin-induced thrombocytopenia: is there enough evidence? PMID- 20023182 TI - How do we bridge the gap? PMID- 20023183 TI - Neuraxial blockade in patients with spinal stenosis: between a rock and a hard place. PMID- 20023185 TI - "Dry" sevoflurane and the falling sky (or lack thereof). PMID- 20023186 TI - False confidences in preoperative pregnancy testing. PMID- 20023187 TI - Estimation of hemodynamic parameters by arterial waveform: available technologies. PMID- 20023188 TI - Double-lumen tubes: which lumen for the gum elastic bougie? PMID- 20023189 TI - Banana blues. PMID- 20023190 TI - Thromboelastometry to guide recombinant activated factor VII therapy for postoperative refractory intracranial bleeding. PMID- 20023193 TI - Society for Pediatric Anesthesia/American Academy of Pediatrics/Congenital Cardiac Anesthesia Society: winter meeting review. PMID- 20023194 TI - Regulation of Arabidopsis brassinosteroid signaling by atypical basic helix-loop helix proteins. AB - Basic helix-loop-helix (bHLH) proteins are highly conserved transcription factors critical for cell proliferation and differentiation. Recent studies have implicated bHLH proteins in many plant signaling processes, including brassinosteroid (BR) signaling. Here, we report identification of two families of atypical bHLH proteins capable of modulating BR signaling. We found that activation-tagged bri1 suppressor 1-Dominant (atbs1-D), previously identified as a dominant suppressor of a weak BR receptor mutant bri1-301, was caused by overexpression of a 93-amino acid atypical bHLH protein lacking amino acids critical for DNA binding. Interestingly, atbs1-D only suppresses weak BR mutants, while overexpression of a truncated ATBS1 lacking the basic motif also rescues bri1-301, suggesting that ATBS1 likely stimulates BR signaling by sequestering negative BR signaling components. A yeast two-hybrid screen using ATBS1 as bait discovered four ATBS1-Interacting Factors (AIFs) that are members of another atypical bHLH protein subfamily. AIF1 exhibits an overlapping expression pattern with ATBS1 and its homologs and interacts with ATBS1 in vitro and in vivo. AIF1 overexpression nullifies the suppressive effect of atbs1-D on bri1-301 and results in dwarf transgenic plants resembling BR mutants. By contrast, silencing of AIF1 partially suppressed the bri1-301 phenotype. Our results suggested that plants use these atypical bHLH proteins to regulate BR signaling. PMID- 20023195 TI - Class I alpha-mannosidases are required for N-glycan processing and root development in Arabidopsis thaliana. AB - In eukaryotes, class I alpha-mannosidases are involved in early N-glycan processing reactions and in N-glycan-dependent quality control in the endoplasmic reticulum (ER). To investigate the role of these enzymes in plants, we identified the ER-type alpha-mannosidase I (MNS3) and the two Golgi-alpha-mannosidase I proteins (MNS1 and MNS2) from Arabidopsis thaliana. All three MNS proteins were found to localize in punctate mobile structures reminiscent of Golgi bodies. Recombinant forms of the MNS proteins were able to process oligomannosidic N glycans. While MNS3 efficiently cleaved off one selected alpha1,2-mannose residue from Man(9)GlcNAc(2), MNS1/2 readily removed three alpha1,2-mannose residues from Man(8)GlcNAc(2). Mutation in the MNS genes resulted in the formation of aberrant N-glycans in the mns3 single mutant and Man(8)GlcNAc(2) accumulation in the mns1 mns2 double mutant. N-glycan analysis in the mns triple mutant revealed the almost exclusive presence of Man(9)GlcNAc(2), demonstrating that these three MNS proteins play a key role in N-glycan processing. The mns triple mutants displayed short, radially swollen roots and altered cell walls. Pharmacological inhibition of class I alpha-mannosidases in wild-type seedlings resulted in a similar root phenotype. These findings show that class I alpha-mannosidases are essential for early N-glycan processing and play a role in root development and cell wall biosynthesis in Arabidopsis. PMID- 20023196 TI - Mutations of an alpha1,6 mannosyltransferase inhibit endoplasmic reticulum associated degradation of defective brassinosteroid receptors in Arabidopsis. AB - Asn-linked glycans, or the glycan code, carry crucial information for protein folding, transport, sorting, and degradation. The biochemical pathway for generating such a code is highly conserved in eukaryotic organisms and consists of ordered assembly of a lipid-linked tetradeccasaccharide. Most of our current knowledge on glycan biosynthesis was obtained from studies of yeast asparagine linked glycosylation (alg) mutants. By contrast, little is known about biosynthesis and biological functions of N-glycans in plants. Here, we show that loss-of-function mutations in the Arabidopsis thaliana homolog of the yeast ALG12 result in transfer of incompletely assembled glycans to polypeptides. This metabolic defect significantly compromises the endoplasmic reticulum-associated degradation of bri1-9 and bri1-5, two defective transmembrane receptors for brassinosteroids. Consequently, overaccumulated bri1-9 or bri1-5 proteins saturate the quality control systems that retain the two mutated receptors in the endoplasmic reticulum and can thus leak out of the folding compartment, resulting in phenotypic suppression of the two bri1 mutants. Our results strongly suggest that the complete assembly of the lipid-linked glycans is essential for successful quality control of defective glycoproteins in Arabidopsis. PMID- 20023199 TI - Collective efficacy in the school context: does it help explain victimization and bullying among Greek primary and secondary school students? AB - Collective efficacy, defined as informal social controls that operate under social norms of trust, is an emerging theoretical concept that has been applied to explain violence rates in neighborhoods, affiliation with deviant peers, partner violence, and adolescent delinquency. This study employed a multilevel design to examine the association between collective efficacy at the class-level and individual-level bullying perpetration and victimization using survey data from 1,729 Greek students, aged 11 to 14 years. School class collective efficacy was defined as cohesion and trust among class members combined with their willingness to intervene in the case of aggressive or bullying incidents. Our findings indicate that individual-level victimization is more frequent in classes with lower levels of collective efficacy. We conclude that the notion of collective efficacy might also prove useful in explaining bullying involvement. PMID- 20023197 TI - Ethylene interacts with abscisic acid to regulate endosperm rupture during germination: a comparative approach using Lepidium sativum and Arabidopsis thaliana. AB - The micropylar endosperm cap covering the radicle in the mature seeds of most angiosperms acts as a constraint that regulates seed germination. Here, we report on a comparative seed biology study with the close Brassicaceae relatives Lepidium sativum and Arabidopsis thaliana showing that ethylene biosynthesis and signaling regulate seed germination by a mechanism that requires the coordinated action of the radicle and the endosperm cap. The larger seed size of Lepidium allows direct tissue-specific biomechanical, biochemical, and transcriptome analyses. We show that ethylene promotes endosperm cap weakening of Lepidium and endosperm rupture of both species and that it counteracts the inhibitory action of abscisic acid (ABA) on these two processes. Cross-species microarrays of the Lepidium micropylar endosperm cap and the radicle show that the ethylene-ABA antagonism involves both tissues and has the micropylar endosperm cap as a major target. Ethylene counteracts the ABA-induced inhibition without affecting seed ABA levels. The Arabidopsis loss-of-function mutants ACC oxidase2 (aco2; ethylene biosynthesis) and constitutive triple response1 (ethylene signaling) are impaired in the 1-aminocyclopropane-1-carboxylic acid (ACC)-mediated reversion of the ABA induced inhibition of seed germination. Ethylene production by the ACC oxidase orthologs Lepidium ACO2 and Arabidopsis ACO2 appears to be a key regulatory step. Endosperm cap weakening and rupture are promoted by ethylene and inhibited by ABA to regulate germination in a process conserved across the Brassicaceae. PMID- 20023198 TI - Arabidopsis formin3 directs the formation of actin cables and polarized growth in pollen tubes. AB - Cytoplasmic actin cables are the most prominent actin structures in plant cells, but the molecular mechanism underlying their formation is unknown. The function of these actin cables, which are proposed to modulate cytoplasmic streaming and intracellular movement of many organelles in plants, has not been studied by genetic means. Here, we show that Arabidopsis thaliana formin3 (AFH3) is an actin nucleation factor responsible for the formation of longitudinal actin cables in pollen tubes. The Arabidopsis AFH3 gene encodes a 785-amino acid polypeptide, which contains a formin homology 1 (FH1) and a FH2 domain. In vitro analysis revealed that the AFH3 FH1FH2 domains interact with the barbed end of actin filaments and have actin nucleation activity in the presence of G-actin or G actin-profilin. Overexpression of AFH3 in tobacco (Nicotiana tabacum) pollen tubes induced excessive actin cables, which extended into the tubes' apices. Specific downregulation of AFH3 eliminated actin cables in Arabidopsis pollen tubes and reduced the level of actin polymers in pollen grains. This led to the disruption of the reverse fountain streaming pattern in pollen tubes, confirming a role for actin cables in the regulation of cytoplasmic streaming. Furthermore, these tubes became wide and short and swelled at their tips, suggesting that actin cables may regulate growth polarity in pollen tubes. Thus, AFH3 regulates the formation of actin cables, which are important for cytoplasmic streaming and polarized growth in pollen tubes. PMID- 20023200 TI - Intimate partner aggression perpetrated and sustained by male Afghanistan, Iraq, and Vietnam veterans with and without posttraumatic stress disorder. AB - Veterans with posttraumatic stress disorder (PTSD) consistently evidence higher rates of intimate partner aggression perpetration than veterans without PTSD, but most studies have examined rates of aggression among Vietnam veterans several years after their deployment. The primary aim of this study was to examine partner aggression among male Afghanistan or Iraq veterans who served during Operation Enduring Freedom (OEF) and Operation Iraqi Freedom (OIF) and compare this aggression to that reported by Vietnam veterans with PTSD. Three groups were recruited, OEF/OIF veterans with PTSD (n = 27), OEF/OIF veterans without PTSD (n = 31), and Vietnam veterans with PTSD (n = 28). Though only a few comparisons reached significance, odds ratios suggested that male OEF/OIF veterans with PTSD were approximately 1.9 to 3.1 times more likely to perpetrate aggression toward their female partners and 1.6 to 6 times more likely to report experiencing female perpetrated aggression than the other two groups. Significant correlations among reports of violence perpetrated and sustained suggested many men may have been in mutually violent relationships. Taken together, these results suggest that partner aggression among Iraq and Afghanistan veterans with PTSD may be an important treatment consideration and target for prevention. PMID- 20023201 TI - Origin of ovarian cancer may have implications for screening. PMID- 20023202 TI - Ductal carcinoma in situ: should the name be changed? PMID- 20023203 TI - StatBite: Relative survival and incidence rates: ductal carcinoma in situ. PMID- 20023204 TI - Mammography screening: after the storm, calls for more personalized approaches. PMID- 20023205 TI - Bile alcohols function as the ligands of membrane-type bile acid-activated G protein-coupled receptor. AB - TGR5 is a G protein-coupled receptor that is activated by bile acids, resulting in an increase in cAMP levels and the subsequent modulation of energy expenditure in brown adipose tissue and muscle. Therefore, the development of a TGR5-specific agonist could lead to the prevention and treatment of various metabolic disorders related to obesity. In the present study, we evaluated the ability of bile alcohols, which are structurally and physiologically similar to bile acids and are produced as the end products of cholesterol catabolism in evolutionarily primitive vertebrates, to act as TGR5 agonists. In a cell-based reporter assay and a cAMP production assay performed in vitro, most bile alcohols with a side chain containing hydroxyl group(s) were highly efficacious agonists for TGR5 comparable to its most potent ligand in the naturally occurring bile acid, lithocholic acid. However, the abilities of the bile alcohols to activate TGR5 varied with the position and number of the hydroxyl substituent in the side chain. Additionally, the conformation of the steroidal nucleus of bile alcohols is also important for its activity as a TGR5 agonist. Thus, we have provided new insights into the structure-activity relationships of bile alcohols as TGR5 agonists. PMID- 20023206 TI - FAT/CD36 is localized in sarcolemma and in vesicle-like structures in subsarcolemma regions but not in mitochondria. AB - The primary aim of the present study was to investigate in which cellular compartments fatty acid trans-locase CD36 (FAT/CD36) is localized. Intact and fully functional skeletal muscle mitochondria were isolated from lean and obese female Zucker rats and from 10 healthy male individuals. FAT/CD36 could not be detected in the isolated mitochondria, whereas the mitochondrial marker F(1)ATPase-beta was clearly detected using immunoblotting. Lack of markers for other membrane structures indicated that the mitochondria were not contaminated with membranes known to contain FAT/CD36. In addition, fluorescence immunocytochemistry was performed on single muscle fibers dissected from soleus muscle of lean and obese Zucker rats and from the vastus lateralis muscle from humans. Costaining against FAT/CD36 and MitoNEET clearly show that FAT/CD36 is highly present in sarcolemma and it also associates with some vesicle-like intracellular compartments. However, FAT/CD36 protein was not detected in mitochondrial membranes, supporting the biochemical findings. Based on the presented data, FAT/CD36 seems to be abundantly expressed in sarcolemma and in vesicle-like structures throughout the muscle cell. However, FAT/CD36 is not present in mitochondria in rat or human skeletal muscle. Thus, the functional role of FAT/CD36 in lipid transport seems primarily to be allocated to the plasma membrane in skeletal muscle. PMID- 20023207 TI - Effect of neutrophil adhesion on the mechanical properties of lung microvascular endothelial cells. AB - Neutrophil adhesion to pulmonary microvascular endothelial cells (ECs) initiates intracellular signaling, resulting in remodeling of F-actin cytoskeletal structure of ECs. The present study determined the mechanical properties of ECs and the changes induced by neutrophil adhesion by atomic force microscopy. The elastic moduli of ECs were compared before neutrophils were present, as soon as neutrophil adhesion was detected, and 1 minute later. ECs that were adjacent to those with adherent neutrophils were also evaluated. Neutrophil adhesion induced a decrease in the elastic moduli in the 6.25-MUm rim of ECs surrounding adherent neutrophils as soon as firmly adherent neutrophils were detected, which was transient and lasted less than 1 minute. Adjacent ECs developed an increase in stiffness that was significant in the central regions of these cells. Intercellular adhesion molecule-1 crosslinking did not induce significant changes in the elastic modulus of ECs in either region, suggesting that crosslinking intercellular adhesion molecule-1 is not sufficient to induce the observed changes. Our results demonstrate that neutrophil adhesion induces regional changes in the stiffness of ECs. PMID- 20023208 TI - Religion as attachment: normative processes and individual differences. AB - The authors review findings from the psychology of religion showing that believers' perceived relationships with God meet the definitional criteria for attachment relationships. They also review evidence for associations between aspects of religion and individual differences in interpersonal attachment security and insecurity. They focus on two developmental pathways to religion. The first is a "compensation" pathway involving distress regulation in the context of insecure attachment and past experiences of insensitive caregiving. Research suggests that religion as compensation might set in motion an "earned security" process for individuals who are insecure with respect to attachment. The second is a "correspondence" pathway based on secure attachment and past experiences with sensitive caregivers who were religious. The authors also discuss conceptual limitations of a narrow religion-as-attachment model and propose a more inclusive framework that accommodates concepts such as mindfulness and "nonattachment" from nontheistic religions such as Buddhism and New Age spirituality. PMID- 20023209 TI - Religiousness as a cultural adaptation of basic traits: a five-factor model perspective. AB - Individual differences in religiousness can be partly explained as a cultural adaptation of two basic personality traits, Agreeableness and Conscientiousness. This argument is supported by a meta-analysis of 71 samples (N = 21,715) from 19 countries and a review of the literature on personality and religion. Beyond variations in effect magnitude as a function of moderators, the main personality characteristics of religiousness (Agreeableness and Conscientiousness) are consistent across different religious dimensions, contexts (gender, age, cohort, and country), and personality measures, models, and levels, and they seem to predict religiousness rather than be influenced by it. The copresence of Agreeableness and Conscientiousness sheds light on other explanations of religiousness, its distinctiveness from related constructs, its implications for other domains, and its adaptive functions. PMID- 20023210 TI - The truth about the truth: a meta-analytic review of the truth effect. AB - Repetition has been shown to increase subjective truth ratings of trivia statements. This truth effect can be measured in two ways: (a) as the increase in subjective truth from the first to the second encounter (within-items criterion) and (b) as the difference in truth ratings between repeated and other new statements (between-items criterion). Qualitative differences are assumed between the processes underlying both criteria. A meta-analysis of the truth effect was conducted that compared the two criteria. In all, 51 studies of the repetition induced truth effect were included in the analysis. Results indicate that the between-items effect is larger than the within-items effect. Moderator analyses reveal that several moderators affect both effects differentially. This lends support to the notion that different psychological comparison processes may underlie the two effects. The results are discussed within the processing fluency account of the truth effect. PMID- 20023211 TI - Teaching pharmacology in an innovative medical curriculum: challenges of integration, technology, and future training. PMID- 20023212 TI - Antibody-array interaction mapping, a new method to detect protein complexes applied to the discovery and study of serum amyloid P interactions with kininogen in human plasma. AB - Protein-protein interactions are fundamentally important in biological processes, but the existing analytical tools have limited ability to sensitively and precisely measure the dynamic composition of protein complexes in biological samples. We report here the development of antibody-array interaction mapping (AAIM) to address that need. We used AAIM to probe interactions among a set of 48 proteins in serum and found several known interactions as well potentially novel interactions, including multiprotein clusters of interactions. A novel interaction initially identified between the innate immune system protein C reactive protein and the inflammatory protein kininogen (KNG) was confirmed in subsequent experiments to involve serum amyloid P instead of its highly related family member, C-reactive protein. AAIM was used in a variety of formats to further study this interaction. In vitro studies confirmed the ability of the purified proteins to interact and revealed a zinc dependence of the interaction. Studies using plasma samples collected longitudinally following a controlled myocardial infarction revealed no consistent changes in the serum amyloid P-KNG interaction levels but consistent changes in KNG activation and interactions with plasma prekallikrein. These results demonstrate a versatile platform for measuring the dynamic composition of protein complexes in biological samples that should have value for studies of normal and disease-related signaling networks, multiprotein clusters, or enzymatic cascades. PMID- 20023213 TI - Distinct roles for LFA-1 affinity regulation during T-cell adhesion, diapedesis, and interstitial migration in lymph nodes. AB - During the course of homing to lymph nodes (LNs), T cells undergo a multistep adhesion cascade that culminates in a lymphocyte function-associated antigen 1 (LFA-1)-dependent firm adhesion to the luminal surface of high endothelial venules (HEVs). The importance of LFA-1 affinity regulation in supporting T-cell arrest on HEVs has been well established, however, its importance in the postadhesion phase, which involves intraluminal crawling and diapedesis to the extravascular space, remains elusive. Here we have shown that LFA-1 affinity needs to be appropriately regulated to support these essential steps in the homing cascade. Genetically engineered T cells that were unable to properly down regulate LFA-1 affinity underwent enhanced, chemokine-independent arrest in HEVs but showed perturbed intravascular crawling to transmigration sites and compromised diapedesis across HEVs. By contrast, the extravascular migration of T cells was insensitive to the affinity-enhancing LFA-1 mutation. These results highlight the requirement for balanced LFA-1 affinity regulation in intravascular and transvascular, but not extravascular, T-cell migration in LNs. PMID- 20023214 TI - alphaB-crystallin regulation of angiogenesis by modulation of VEGF. AB - alphaB-crystallin is a chaperone belonging to the small heat shock protein family. Herein we show attenuation of intraocular angiogenesis in alphaB crystallin knockout (alphaB-crystallin(-/-)) mice in 2 models of intraocular disease: oxygen-induced retinopathy and laser-induced choroidal neovascularization. Vascular endothelial growth factor A (VEGF-A) mRNA and hypoxia inducible factor-1alpha protein expression were induced during retinal angiogenesis, but VEGF-A protein expression remained low in alphaB-crystallin(-/ ) retina versus wild-type mice, whereas VEGF-R2 expression was not affected. Both alphaB-crystallin and its phosphorylated serine59 formwere expressed, and immunoprecipitation revealed alphaB-crystallin binding to VEGF-A but not transforming growth factor-beta in cultured retinal pigment epithelial (RPE) cells. alphaB-crystallin and VEGF-A are colocalized in the endoplasmic reticulum in RPE cells under chemical hypoxia. alphaB-crystallin(-/-) RPE showed low VEGF-A secretion under serum-starved conditions compared with wild-type cells. VEGF-A is polyubiquitinated in control and alphaB-crystallin siRNA treated RPE; however, mono-tetra ubiquitinated VEGF-A increases with alphaB-crystallin knockdown. Endothelial cell apoptosis in newly formed vessels was greater in alphaB crystallin(-/-) than wild-type mice. Proteasomal inhibition in alphaB-crystallin( /-) mice partially restores VEGF-A secretion and angiogenic phenotype in choroidal neovascularization. Our studies indicate an important role for alphaB crystallin as a chaperone for VEGF-A in angiogenesis and its potential as a therapeutic target. PMID- 20023217 TI - Yes, Virginia, there will be healthcare reform. PMID- 20023215 TI - Rituximab plus concurrent infusional EPOCH chemotherapy is highly effective in HIV-associated B-cell non-Hodgkin lymphoma. AB - Rituximab plus intravenous bolus chemotherapy is a standard treatment for immunocompetent patients with B-cell non-Hodgkin lymphoma (NHL). Some studies have suggested that rituximab is associated with excessive toxicity in HIV associated NHL, and that infusional chemotherapy may be more effective. We performed a randomized phase 2 trial of rituximab (375 mg/m(2)) given either concurrently before each infusional etoposide, vincristine, doxorubicin, cyclophosphamide, and prednisone (EPOCH) chemotherapy cycle or sequentially (weekly for 6 weeks) after completion of all chemotherapy in HIV-associated NHL. EPOCH consisted of a 96-hour intravenous infusion of etoposide, doxorubicin, and vincristine plus oral prednisone followed by intravenous bolus cyclophosphamide given every 21 days for 4 to 6 cycles. In the concurrent arm, 35 of 48 evaluable patients (73%; 95% confidence interval, 58%-85%) had a complete response. In the sequential arm, 29 of 53 evaluable patients (55%; 95% confidence interval, 41% 68%) had a complete response. The primary efficacy endpoint was met for the concurrent arm only. Toxicity was comparable in the 2 arms, although patients with a baseline CD4 count less than 50/microL had a high infectious death rate in the concurrent arm. We conclude that concurrent rituximab plus infusional EPOCH is an effective regimen for HIV-associated lymphoma. PMID- 20023216 TI - CCR6 ligands inhibit HIV by inducing APOBEC3G. AB - We have identified a post-entry CCR6-dependent mechanism of inhibition of HIV occurring at an early stage of infection mediated by the induction of the host restriction factor apolipoprotein B mRNA-editing enzyme-catalytic polypeptide like 3G (APOBEC3G). We observed induction of APOBEC3G expression only in CCR6(+) cells but not in cells treated with the G inhibitory (Gi) pathway inhibitor pertussis toxin. CCR6 is highly expressed on peripheral blood CD4(+)CCR5(+) memory T cells and by 2 populations of CD4(+) T cells within the gut, alpha4beta7(+) and T helper type 17, that have been implicated in cell-to-cell spread of HIV and enhanced restoration of CD4(+) T cells within gut-associated lymphoid tissue, respectively. This novel CCR6-mediated mechanism of inhibition allows the identification of pathways that induce intrinsic immunity to HIV, which could be useful in devising novel therapeutics that selectively target CCR6(+) cells. PMID- 20023218 TI - Protecting patients and maintaining professional sovereignty in the midst of intrusive government change. PMID- 20023219 TI - Eliminating barriers for patients with limited English proficiency. AB - The increasing number of patients with limited English proficiency (LEP) in the United States is an important issue for patients, physicians, federal and state government agencies, and insurance carriers. Physicians can actively address this healthcare barrier by becoming familiar with established state and federal guidelines and determining when appropriate linguistic services are required for their LEP patient populations. Physicians can ensure that appropriate care is administered to LEP patients by developing and monitoring plans that include training staff to identify and work with LEP patients, providing language assistance to LEP patients, and notifying LEP patients of special services. PMID- 20023220 TI - Carpal tunnel syndrome: ultrasonographic imaging and pathologic mechanisms of median nerve compression. AB - Median nerve compression is a well-known cause of carpal tunnel syndrome (CTS). Yet, reasons why the most common idiopathic form of CTS develops in certain individuals are not well understood. To further understand the compressive mechanisms at work in CTS development, the authors used ultrasonographic imaging of the median nerve to evaluate 2 patients with CTS. Findings were compared to those of 2 control subjects who did not have CTS. In the patients who had CTS, the transverse carpal ligament was pulled taut by thenar muscle contraction as the flexor tendons tightened, compressing the median nerve between the ligament and tendons. No such compression was observed with the control subjects. Thus, a pathologic mechanism of median nerve compression was confirmed in the patients with CTS. Demonstration of such pathologic mechanisms during prehensile hand movement may improve understanding of how to treat patients with CTS and prevent nerve injury. PMID- 20023221 TI - Musculoskeletal dysfunction and drop foot: diagnosis and management using osteopathic manipulative medicine. AB - Drop foot arises from dysfunction within the anatomic, muscular, or neurologic aspects of the lower extremity. The authors describe a patient with drop foot who had a compressed common peroneal nerve caused by posterior fibular head dysfunction. One 15-minute session of osteopathic manipulative treatment resolved the patient's symptoms. It is important for physicians to use osteopathic manipulative medicine to diagnosis and manage this condition, particularly when it results from fibular head dysfunction. PMID- 20023222 TI - Perception of osteopathic medicine among allopathic physicians in the deep central southern United States. PMID- 20023223 TI - Keeping the osteopathic medical profession parallel and distinctive. PMID- 20023224 TI - Discrimination against DOs alive and well. PMID- 20023225 TI - Cranial palpation pressures used by osteopathy students. PMID- 20023226 TI - Reject influence of pharmaceutical industry. PMID- 20023227 TI - Neurophysiological evaluation of convergent afferents innervating the human esophagus and area of referred pain on the anterior chest wall. AB - Noxious stimuli in the esophagus cause pain that is referred to the anterior chest wall because of convergence of visceral and somatic afferents within the spinal cord. We sought to characterize the neurophysiological responses of these convergent spinal pain pathways in humans by studying 12 healthy subjects over three visits (V1, V2, and V3). Esophageal pain thresholds (Eso-PT) were assessed by electrical stimulation and anterior chest wall pain thresholds (ACW-PT) by use of a contact heat thermode. Esophageal evoked potentials (EEP) were recorded from the vertex following 200 electrical stimuli, and anterior chest wall evoked potentials (ACWEP) were recorded following 40 heat pulses. The fear of pain questionnaire (FPQ) was administered on V1. Statistical data are shown as point estimates of difference +/- 95% confidence interval. Pain thresholds increased between V1 and V3 [Eso-PT: V1-V3 = -17.9 mA (-27.9, -7.9) P < 0.001; ACW-PT: V1 V3 = -3.38 degrees C (-5.33, -1.42) P = 0.001]. The morphology of cortical responses from both sites was consistent and equivalent [P1, N1, P2, N2 complex, where P1 and P2 are is the first and second positive (downward) components of the CEP waveform, respectively, and N1 and N2 are the first and second negative (upward) components, respectively], indicating activation of similar cortical networks. For EEP, N1 and P2 latencies decreased between V1 and V3 [N1: V1-V3 = 13.7 (1.8, 25.4) P = 0.02; P2: V1-V3 = 32.5 (11.7, 53.2) P = 0.003], whereas amplitudes did not differ. For ACWEP, P2 latency increased between V1 and V3 [ 35.9 (-60, -11.8) P = 0.005] and amplitudes decreased [P1-N1: V1-V3 = 5.4 (2.4, 8.4) P = 0.01; P2-N2: 6.8 (3.4, 10.3) P < 0.001]. The mean P1 latency of EEP over three visits was 126.6 ms and that of ACWEP was 101.6 ms, reflecting afferent transmission via Adelta fibers. There was a significant negative correlation between FPQ scores and Eso-PT on V1 (r = -0.57, P = 0.05). These data provide the first neurophysiological evidence of convergent esophageal and somatic pain pathways in humans. PMID- 20023228 TI - Selective deficiency of HIF-1alpha in myeloid cells influences secondary intention wound healing in mouse skin. AB - BACKGROUND: Hypoxia-inducible factor-1 (HIF-1) influences myeloid cell function. In this study we examined the role of myeloid cell HIF-1alpha on wound healing in vivo using a cell-specific knockout (KO) mouse model. MATERIALS AND METHODS: HIF 1alpha KO mice and wild-type (WT) controls received 8 mm full thickness dorsal dermal wounds. Wound dimensions were measured until full closure. Tissue was obtained from 3-day-old wounds for (immuno-)histochemical analysis. Production of interleukin-1beta (IL-1beta) and nitric oxide (NO) in response to lipopolysaccharide (LPS) and/or desferrioxamine (DFX) was examined in vitro. RESULTS: Early wound closure occurred significantly faster in HIF-1alpha KO mice than in WT mice. Wounds of KO mice contained similar numbers of neutrophils and macrophages, but more activated keratinocytes, consistent with accelerated re epithelialization. Interestingly, while LPS and LPS+DFX elicited a similar IL 1beta response in macrophages from the 2 mouse types, NO production was blunted in HIF-1alpha KO macrophages. CONCLUSION: Absence of HIF-1alpha in myeloid cells accelerates the early phase of secondary intention wound healing in vivo. This may be associated with a deficient ability of myeloid cells to initiate an appropriate NO production response. Pharmacologic modulators of HIF-1alpha should be explored in situations with abnormal wound healing. PMID- 20023229 TI - Efficient liver-directed gene transfer by in situ generation of retroviral vector from adenoviral templates. AB - To improve liver-directed retroviral-mediated gene transfer, we injected C57/BL10 mice intravenously with three adenoviral vectors encoding retroviral vector genome and structural components: AdGagPol expressing the respective structural genes of Moloney murine leukaemia virus, Ad10A1Env expressing the 10A1 envelope protein of 10A1-MuLV, and AdLEIN, encoding the LEIN retrovirus genome, expressing green fluorescence protein (eGFP) and the neomycin resistance gene. MATERIALS AND METHODS: The extent of eGFP expression was determined after 1 and 15 weeks by fluorescence microscopy and FACS analysis. Proviral integration was determined by a novel PCR-based technique. RESULTS: Hepatocytes infected with all three Ad vectors generated LEIN retrovirus after one week and in situ transduction of neighbouring cells resulted in stable proviral integration associated with eGFP expression ranging from 4.3% to 20.5% in different liver cell populations 15 weeks post-infection. CONCLUSION: Hybrid adeno-retroviral vectors can be efficiently used to improve the efficiency of retroviral-mediated gene transfer to the liver. PMID- 20023230 TI - Phenotypic correction of Age-associated functional decline in murine immune cells by Thymax, a thymic extract. AB - BACKGROUND: We have recently demonstrated that Thymax, a gross thymic extract, induces the functional activity of human dendritic cells (DCs) in vitro. In this study, the role of Thymax in phenotypic correction of age-associated functional decline in immune cells in mice was evaluated. MATERIALS AND METHODS: C57BL/6 mice (13 months old) were treated with Thymax orally (20% v/v) for 4 weeks. Different splenic cell types, dendritic cells (DCs), B-cells, T-cells and natural killer (NK) cells, were analyzed using flow cytometry. RESULTS: Treatment with Thymax resulted in: i) a significant increase in the percentages of DCs (1.6 fold), B-cells (7-fold) and T-cells (5-fold) over the control (p<0.05); ii) an increase in the percentages of activation markers (CD25 and CD69) of CD4(+) and CD8(+) T-cells; and iii) an enhancement in NK activity. Thymax showed no adverse side-effects. CONCLUSION: Thymax might have a role in reversing immune dysfunction in the elderly. PMID- 20023231 TI - Inhibiting platelet-derived growth factor beta reduces Ewing's sarcoma growth and metastasis in a novel orthotopic human xenograft model. AB - BACKGROUND: Despite aggressive therapy, Ewing's sarcoma (ES) patients have a poor five-year overall survival of only 20-40%. Pulmonary metastasis is the most common form of demise in these patients. The pathogenesis of pulmonary metastasis is poorly understood and few orthotopic models exist that allow study of spontaneous pulmonary metastasis in ES. MATERIALS AND METHODS: We have developed a novel orthotopic xenograft model in which spontaneous pulmonary metastases develop. While the underlying biology of ES is incompletely understood, in addition to the EWS-FLI-1 mutation, it is known that platelet-derived growth factor receptor beta (PDGFR-beta) is highly expressed in ES. Hypothesizing that PDGFR-beta expression is indicative of a specific role for this receptor protein in ES progression, the effect of PDGFR-beta inhibition on ES growth and metastasis was assessed in this novel orthotopic ES model. RESULTS: Silencing PDGFR-beta reduced spontaneous growth and metastasis in ES. CONCLUSION: Preclinical therapeutically relevant findings such as these may ultimately lead to new treatment initiatives in ES. PMID- 20023232 TI - Chemotherapy in canine acute megakaryoblastic leukemia: a case report and review of the literature. AB - Acute myeloid leukemia (AML) in dogs is a rare disease with poor prognosis. In most subjects, palliative treatment or euthanasia is performed. A 3.5-year-old male castrated labrador with AML-M7, which was treated with induction polychemotherapy (8 cycles) using vincristine (0.5 mg/m(2)/cycle), daunorubicin (20 mg/m(2)/cycle), cytosine arabinoside (ARA-C, 100 mg/m(2)/cycle) and prednisolone (1 mg/kg/day) is reported. Treatment was well tolerated and complete remission was achieved. Postinduction chemotherapy consisted of ARA-C, daunorubicin and prednisolone. After 3, 5 and 18 months, the subject relapsed. Each relapse was treated with ARA-C (up to 1,000 mg/m(2)) and etoposide or daunorubicin. Again, no severe side-effects occurred and the disease was controlled, with 37 chemotherapy-cycles (ARA-C, 3 x 1,000 mg/m(2)/cycle), for 24 months. Based on a literature-search, this is the first report documenting a long term response of canine AML, probably resulting from the high-dose ARA-C. Clinical trials using high-dose ARA-C are now required to confirm antileukemic efficacy in canine leukemias. PMID- 20023233 TI - Role of nitrites in the genesis of adenocarcinoma associated with Barrett's esophagus. AB - BACKGROUND: Barrett's esophagus (BE) is one of the complications of gastroesophageal reflux disease (GERD) and a premalignant condition. It consists of a process of replacement of the squamous epithelium of the esophagus by intestinal columnar epithelium containing goblet cells, known as specialized intestinal metaplasia with goblet cells, and several factors have been related to its pathogenesis. The objective of this study was to evaluate an experimental model of duodenogastroesophageal reflux and the effect of ingestion of sodium nitrite solution on the genesis of adenocarcinoma associated with Barrett's esophagus. MATERIALS AND METHODS: Sixty male Wistar rats were divided into four groups. Twenty were not submitted to surgery and served as controls (10 animals ingesting only water and 10 ingesting water plus a solution of sodium nitrite), while the remaining 40 animals were submitted to side-to-side duodenogastroesophageal anastomosis (20 animals ingesting only water and 20 ingesting water plus the sodium nitrite solution). The Vienna classification for dysplasia and adenocarcinoma was used in the analysis of results. RESULTS: After 42 weeks of observation, Barrett's esophagus was found in 26.3% (5/19) of the animals submitted to surgery that had not ingested nitrites compared to 72.3% (13/18) of the animals in the group submitted to surgery and given nitrites. Six cases of adenocarcinoma (33.3%) were also found in this latter group. Barrett's esophagus was not found in any of the animals that were not submitted to surgery. Categories 2, 3 and 5 of the Vienna classification were only found in the animals submitted to surgery that also received sodium nitrite (66.7%). CONCLUSION: The ingestion of sodium nitrite associated with duodenogastroesophageal reflux plays an important role in the genesis of adenocarcinoma associated with Barrett's esophagus. PMID- 20023234 TI - The frequency of glassy cells in Barrett's mucosa: a study in Baboons. AB - BACKGROUND: Glands with glassy cells (GGCs) were recently found in 1.8% of patients showing Barrett's mucosa in esophageal biopsies. Similar GGCs were more recently detected in a baboon having glandulo-metaplastic esophageal mucosa (GMEM). The aim was to assess the frequency of baboons with GMEM having GGCs. MATERIALS AND METHODS: GGCs were sought in 68 consecutive baboons having GMEM. Sections were stained with H&E, and with alcian blue (pH 2.5) to detect sialomucins in goblet cells (a marker of Barrett's mucosa in GMEM). RESULTS: Two out of the 68 baboons with Barrett's mucosa (2.9%) showed GGCs. CONCLUSION: In similarity to humans, the Barrett's mucosa in baboons may show GGCs. Although the significance of GGCs in baboons (and in humans) remains poorly understood, their presence might not be a fortuitous event but may be linked to the molecular events leading to the development of intestinal metaplasia in Barrett's mucosa, a known pre-neoplastic mucosal change in the distal esophagus in humans. PMID- 20023235 TI - The effects of alginate encapsulation on NIT-1 insulinoma cells: viability, growth and insulin secretion. AB - Transplantation of microencapsulated insulin-secreting cells is proposed as a promising therapy for the treatment of type I diabetes mellitus. In recent years, important advances have been made in the field of immunoisolation and many studies have shown that alginate provides some major advantages for encapsulation over other systems. Since it is known that the extracellular matrix influences the behaviour of encapsulated cells, the aim of the present work has been to study the consequences of encapsulation on some cell functions. For this purpose, cell growth and dynamics of insulin release of NIT-1 cells entrapped in alginate capsules compared with those exhibited by free NIT-1 cells were investigated by means of growth curves, assays, Trypan blue staining and ELISA test. All investigations performed allowed us to conclude that alginate-entrapped NIT-1 cells maintain their growth features and secretory functions although with some important differences. In particular, alginate encapsulation affects the cellular growth profile and causes the lost of time dependence of insulin secretion profile. PMID- 20023236 TI - Imbalance of tumor suppression genes expression following rat tongue carcinogenesis induced by 4-nitroquinoline 1-oxide. AB - This study was undertaken to investigate, by immunohistochemistry, the expression of some tumor suppressor genes such as p16, p21 and Retinoblastoma (Rb) during 4 Nitroquinoline 1-oxide induced rat tongue carcinogenesis. Male Wistar rats were distributed into three groups of 10 animals each and treated with 50 ppm 4NQO solution through their drinking water for 4, 12 or 20 weeks. Ten animals were used as negative control. Neither histopathological abnormalities were induced in the epithelium after 4 weeks of carcinogen exposure, nor statistically significant differences (p>0.05) in expression of all the tumor suppressor genes were found when compared to the negative control. However, the levels of Rb were increased (p<0.05) in pre-neoplastic lesions at 12 weeks following carcinogen exposure. In well-differentiated squamous cell carcinoma induced after 20 weeks of treatment with 4NQO, p16 and Rb were expressed in some tumor cells. Taken together, the results support the belief that the expression of Rb is closely event-related to malignant transformation and conversion of the oral mucosa, being a reliable biomarker linked to oral cancer pathogenesis. PMID- 20023237 TI - Differential effect of phenothiazines on MRP1 and P-glycoprotein activity. AB - BACKGROUND: Overexpression of ATP-binding cassette (ABC) transporters such as P glycoprotein (P-gp), multidrug resistance-associated protein 1 (MRP1) or breast cancer resistance protein (BCRP) accounts for majority of cases of multidrug resistance (MDR) of cancer cells. MATERIALS AND METHODS: In the present work, the interactions of seven commercially available phenothiazine derivatives, known P glycoprotein inhibitors, with this transporter and MRP1 were compared. By flow cytometry, it was shown that all the drugs increased the accumulation of rhodamine 123 in the P-gp-overexpressing lymphoma cell line L5178 MDR. On the other hand, phenothiazine derivatives stimulated MRP1-mediated efflux of fluorescent probe (BCPCF) out of human erythrocytes. RESULTS: In this way, these phenothiazine derivatives were identified as a group of atypical MDR modulators that differently interact with P-gp (as inhibitors) and MRP1 (as stimulators). CONCLUSION: This observation clearly shows that the activity of all new modulators should be tested for their effects towards different ABC transporters as a standard procedure. PMID- 20023238 TI - Associations of -374T/A polymorphism of receptor for advanced glycation end products (RAGE) gene in Turkish diabetic and non-diabetic patients with coronary artery disease. AB - BACKGROUND: In this study we aimed to determine the possible risks for the development of coronary artery disease (CAD) in diabetic (DM(+)) and non-diabetic (DM(-)) patients according to the -374T/A polymorphism of the receptor for advanced glycation end products (RAGE) gene which affects the function of RAGE itself. MATERIALS AND METHODS: This study was carried out in 52 non-diabetic and 62 diabetic patients with CAD, and 55 CAD-free, healthy volunteers as controls. The A-T transversion polymorphism at position -374 in the promotor region of the RAGE gene was analyzed by polymerase chain reaction (PCR)-restriction fragment length polymorphism (RFLP) techniques. RESULTS: The -374T/A AA genotype frequency was statistically higher in the whole patient group when compared with the control group (p=0.034), and statistically higher in the DM(+) group when compared with the control group (p=0.003). Homozygosity for the -374A allele was found to be higher, but not statistically meaningful, in DM(-) patients (17.3%) when compared with the control group (13.2%). In this study, in contrast with other studies, we found possesion of the A allele to be an independent risk factor in CAD in patients with diabetes mellitus. CONCLUSION: Possesion of the 374A allele may contribute to the CAD in diabetic patients with triggering macrophages by increased levels of AGEs. PMID- 20023240 TI - Chemosensitizing effect of nordihydroguaiaretic acid and its tetra-acetylated derivative on parental and multiresistant TA3 mouse mammary adenocarcinoma cells. AB - BACKGROUND: Multidrug resistance (MDR) continues being the major obstacle for successful anticancer chemotherapy. MATERIALS AND METHODS: The action of nordihydroguaiaretic acid (NDGA) and its tetra-acetylated derivative (NDGATA) on TA3 mouse mammary adenocarcinoma cells and their ability to restore doxorubicin (DOX), cisplatin (CPT) and methotrexate (MTX) sensitivity of the multiresistant variant TA3-MTX-R was examined. RESULTS: Both NDGA and NDGATA synergistically enhanced the cytotoxicity of DOX, CPT and MTX, with a more evident effect in the TA3-MTX-R than in the TA3 cells. NDGATA was more effective than NDGA, as analyzed by the isobologram method. The combination of NDGATA and DOX also reduced the tumor growth rate in mice. Although it did not prolong the median survival time, 30% of mice showed no vestiges of tumor 200 days after implantation with either TA3 or TA3-MTX-R cells. Moreover, NDGA and NDGATA increased the accumulation of DOX and rhodamine (RHO) 123 in both cell lines. CONCLUSION: NDGA and NDGATA are able to chemosensitize tumor cells and combination therapy with NDGATA and DOX is effective at inhibiting tumor growth in mice. PMID- 20023239 TI - Further studies on the frequency and length of the glandulo-metaplastic esophageal mucosa in baboons. AB - BACKGROUND: In previous studies, the length of the glandulo-metaplastic esophageal mucosa (GMEM) at the gastroesophageal junction was assessed in a selected group of baboons. In this study, the length of the GMEM was measured in the entire esophagus in a cohort of unselected adult baboons. MATERIALS AND METHODS: In 15 female baboons, the entire esophagus was removed en bloc at autopsy, from the tongue to the angle of His. No part of the stomach was included. The length of GMEM was measured using a calibrated ocular microscale. RESULTS: GMEM was found in 11 out of the 15 esophagi. The total length of GMEM recorded in the 11 cases was 115 mm (mean 10.5 mm, range 1-45 mm). The mean age for animals with GMEM was 15.5 years (range 7-32 years) and for animals without GMEM was 14.0 years (range 7-20 years); the difference was non-significant (p<0.6). No significant association was found between the length of the GMEM and the age of the animals (p<0.6). CONCLUSION: This study substantiates the notion that GMEM in baboons is a postnatal physiological adaptative process of the esophageal mucosa to daily regurgitation with rumination of gastric juices of low pH. The GMEM apparently progresses upwards, along the esophageal mucosa. The baboon might be an excellent animal model to study the series of histological events that take place in the distal esophagus under the influence of protracted gastroesophageal reflux. PMID- 20023241 TI - Is LOX-1 K167N polymorphism protective for coronary artery disease? AB - BACKGROUND: Human lectin-like oxidized low-density lipoprotein receptor 1 (LOX-1, OLR1) has been identified as a cell surface endocytosis receptor for oxidized low density lipoprotein (oxLDL) on vascular endothelial cells. OxLDLs are avidly ingested by macrophages, resulting in foam cell formation. OxLDLs are also involved in inducing smooth muscle cell migration, proliferation and transformation. A single nucleotide polymorphism K167N (G501C) of the LOX-1 gene results in an amino acid dimorphism (Lys/Asn) at residue 167. Replacement of this Lys residue causes reduced binding and internalization of oxLDL. The purpose of this study was to investigate the effect of the LOX-1 K167N gene polymorphism in Turkish patients with coronary artery disease (CAD). MATERIALS AND METHODS: K167N polymorphism were studied in 91 patients with CAD and 72 healthy controls by the PCR-RFLP method. RESULTS: The frequencies of the KK genotype and the K allele were higher in the CAD group than the controls (p<0.05), while the frequency of the NN genotype was higher in the control group than in the CAD group (p<0.05). It was observed that the decreased CAD risk in patients who had the N allele was reversed by male sex (OR: 0.400 -->0.481) and smoking (OR: 0.400 -->0.949). Although male sex and smoking were lower than other cardiovascular risk factors in patients with the N allele they were higher than other cardiovascular risk factors in patients with the K allele. CONCLUSION: Male sex and smoking decrease the protective effects of the N allele. The adverse effects of the K allele on the CAD risk resulting from the K167N polymorphism appear to be independent of other cardiovascular risk factors. PMID- 20023242 TI - The plant-derived natural compound Flavin 7 attenuates oxidative stress in cultured renal proximal tubule cells. AB - BACKGROUND: Cancer therapies and cancer progression can increase oxidative stress that might account for renal toxicity in cancer patients. Flavin 7 (F7) is a natural polyphenol-containing dietary supplement with potential antioxidant activity. Therefore, it might help to attenuate renal toxicity of chemotherapeutics. MATERIALS AND METHODS: Cultured mouse renal proximal tubule cells were subjected to H(2)O(2)-mediated oxidative stress. Potential antioxidant effects of F7 were assessed by measuring the production of reactive oxygen species (ROS), mitochondrial depolarization and injury (lactate dehydrogenase release as well as trypan blue exclusion) in cells that were pretreated with F7 prior to treatment with H(2)O(2). RESULTS: F7 pretreatment significantly attenuated H(2)O(2)-induced ROS production, mitochondrial depolarization and consequent injury in renal proximal tubule cells. CONCLUSION: F7 supplementation might be beneficial for cancer patients in order to prevent renal toxicity of anticancer drug- or cancer progression-related oxidative stress. PMID- 20023243 TI - Fundic gland cysts in Atp4a-/- mice mimic fundic gland polyps in humans. AB - BACKGROUND: Innumerable fundic gland polyps (FGPs) (that is massive FGP polyposis) may be found in the human stomach. Innumerable fundic gland cysts (FGCs) develop in mice lacking the gene Atp4a (Atp4a((-/-))), that encodes the enzyme ATPase. The aim was to monitor the evolution of FGCs in Atp4a((-/-)) mice. MATERIALS AND METHODS: Six pairs (male-female) of Atp4a((-/-)) mice, 8, 12 and 20 months old, were investigated. Wild-type Atp4a((+/+)) mice were used as controls. RESULTS: In Atp4a((-/-)) female mice, the mucosal thickness increased both at 8 months (943+/-72 mm) (p<0.02) and at 12 months (p<0.0008) and even further at 20 months (p<0.0001). The FGC area was 13.183+/-7.218 mm(2) at 12 months and 73.361+/-50.591 mm(2) at 20 months (p<0.0008). FGCs in Atp4a((-/-)) mice were lined by parietal cells, mucous pit-foveolar cells and chief cells, thus mimicking FGPs in humans. CONCLUSION: Older female Atp4a((-/-)) mice developed innumerable FGCs, a phenomenon very similar to massive FGPs seen in humans. In the light of these similarities, the Atp4a((-/-)) mouse emerges as a suitable animal model to investigate the series of events taking place during the evolution of FGPs in humans. PMID- 20023244 TI - Evaluation of polyurethane dressing with ibuprofen in the management of split thickness skin graft donor sites. AB - BACKGROUND: The authors investigated the possible effect of ibuprofen when included in polyurethane dressing foam in the management of pain and healing related to split-thickness skin graft (STSG) donor sites. The study focused on the use of a foam dressing, Biatain-Ibu, the combination of an absorbent hydrophilic polyurethane foam, Biatain, and the active ingredient ibuprofen as an integral part of the matrix. PATIENTS AND METHODS: A prospective study was conducted from October 2006 to April 2007 and included 40 patients undergoing surgery for any reconstructive purposes with the use of STSG. The patients were divided into two groups in a randomized fashion. In the first group of 20 patients, the donor sites were covered using Biatain-Ibu foam dressing. In the second group of 20 patients, the donor sites were closed intra-operatively with a standard dressing which did not contain any known healing promoting agent. To evaluate the extent and quality of the pain experienced by the patients and to score pain over time, the patients in the study were asked to complete a form containing a visual analogue scale and answer questions on the quality of pain and the way normal daily activities were affected. RESULTS: The combined use of ibuprofen with bio-occlusive dressings accelerated wound healing compared to fine mesh gauze dressings and almost eliminated pain and discomfort in all patients treated. In patients receiving topical ibuprofen, itch did not present a major problem. DISCUSSION: This study demonstrates that the Biatain-Ibu dressing is a useful tool in the management of STSG donor sites by providing an optimal environment for wound healing due to its bio-occlusive properties and by minimizing pain and discomfort. PMID- 20023245 TI - HER2 expression in breast cancer: correlation with endocrine function and psychological status in operable and metastatic breast cancer. AB - BACKGROUND: Node involvement, negative estrogen receptor (ER) and HER2 expression are the main negative prognostic factors for breast cancer. Prolactin (PRL) is involved in the control of breast cancer growth and differentiation. Surgery induced hyperprolactinemia seems to be a positive prognostic factor for operable breast cancer, whereas high PRL levels may predict a poor prognosis in women with metastatic breast cancer. In this study, we evaluated the relation between HER2 expression and PRL blood concentrations in women with metastatic breast cancer women and those whit operable breast cancer patients prior to before and 7 days after surgery. PATIENTS AND METHODS: The study included 50 women with breast cancer, 22 of whom had metastatic disease. HER 2 expression and serum levels of PRL were evaluated by fluorescence in situ hybridization (FISH) method and immunoradiometric assay (IRMA) method, respectively. RESULTS: HER2 expression occurred in 11/28 operable cases and in 8/22 metastatic cases. The percentage of surgery-induced hyperprolactinemia was significantly higher in HER2-negative patients than in those with its expression. Moreover, HER2-positive metastatic cases showed significantly higher mean serum PRL levels than in the negative group. CONCLUSION: These preliminary results show that metastatic cancer-related hyperprolactinemia and lack of surgery-induced hyperprolactinemia are statistically more frequent in HER2-positive patients, thus suggesting a link between PRL endogenous secretion and HER2 expression in breast cancer. PMID- 20023246 TI - Clinical application of autologous three-cellular cultured skin substitutes based on esterified hyaluronic acid scaffold: our experience. AB - BACKGROUND: The aim of this work was to present our experience in the use of autologous three-cellular cultured skin substitutes (CSS). This method represents a surgical alternative in the treatment of a variety of pathologies, including burns, ulcers, giant nevi and tumors. PATIENTS AND METHODS: CSS were obtained from full-thickness skin biopsies collected after enrolment of 11 patients in a clinical trial protocol approved by the local Institutional Review Boards of the 'La Sapienza' University of Rome and registered in clinicaltrials.gov (ID: NCT00718978). CSS consisted of a structure made by a pluristratified epithelial cell surface with melanocytes (ratio 1/20) and a basement of fibroblasts kept together by an esterified hyaluronic acid scaffold that can be surgically manipulated and is gradually reabsorbed after implantation and substituted by the host connectival stroma. RESULTS: At the time of withdrawal of medication, the graft take was comparable to that of autografts, whereas in the follow-up visits, loss of the epithelial layer varied markedly (from 5 to 70%) while fibroblast cellular component growth prevailed. In one patient, there was complete dermal epidermal construct survival. CONCLUSION: Given the anatomical complexity of the skin, we still have a long way to go before we are able to recreate all the cellular and structural characteristics of this organ. Results are, however, gradually improving, as is demonstrated by a successful graft, which was histologically shown to have a three-dimensional structure that maintained its conformation in vivo (epithelium, basement membrane, dermis, subcutis) in one patient. The take of melanocytes improved the final esthetic outcome. PMID- 20023247 TI - Polymorphisms in the promoter region of relaxin-2 and preterm birth: involvement of relaxin in the etiology of preterm birth. AB - BACKGROUND: Circulating relaxin levels have repeatedly been associated with preterm birth (PTB). Our aim was to investigate if mothers carrying promoter single nucleotide polymorphisms (SNPs) in one of the three relaxin genes (RLN1, RLN2, RLN3) are predisposed for PTB. PATIENTS AND METHODS: Maternal DNA from 80 preterm cases (40 very preterm births (24-34 weeks) and 40 moderate preterm (34 36 weeks) and 40 controls (term delivery)) nested in the Danish National Birth Cohort were examined for nine SNPs. RESULTS: Maternal homozygosity of the rarer allele in the relaxin 2 gene (RLN2, rs10115467 and rs4742076) had an increased risk of moderate PTB (odds ratio 4.1 [95% CI 1.4-12] and 8.8 [95% CI 1.03-75] respectively). Only rs10115467 remained significant after correction for multiple testing. CONCLUSION: Women homozygous for prevalent SNPs in the RLN2 gene may have a genetic susceptibility for PTB. PMID- 20023248 TI - A clinical pilot study of lignin--ascorbic acid combination treatment of herpes simplex virus. AB - Antiviral drugs as well as natural remedies have been used to reduce symptoms and the rate of recurrences of herpes simplex virus type 1 (HSV-1) infection, a common disease. To evaluate anti-HSV-1 activity of a pine cone lignin and ascorbic acid treatment, a clinical pilot study was carried out. Forty-eight healthy patients of both genders between 4 and 61 years old (mean: 31+/-16 years), with active lesions of HSV-1, took part in the study. According to the HSV-1 stage at the presentation, the patients were classified into the prodromic (16 patients), erythema (11 patients), papule edema (1 patient), vesicle/pustule (13 patients) and ulcer stages (7 patients). One mg of lignin-ascorbic acid tablet or solution was orally administered three times daily for a month. Clinical evaluations were made daily the first week and at least three times a week during the second week after the onset and every six months during the subsequent year to identify recurrence episodes. The patients who began the lignin-ascorbic acid treatment within the first 48 hours of symptom onset did not develop HSV-1 characteristic lesions, whereas those patients who began the treatment later experienced a shorter duration of cold sore lesions and a decrease in the symptoms compared with previous episodes. The majority of the patients reported the reduction in the severity of symptoms and the reduction in the recurrence episodes after the lignin-ascorbic acid treatment compared with previous episodes, suggesting its possible applicability for the prevention and treatment of HSV-1 infection. PMID- 20023249 TI - Incidence of arm lymphoedema following sentinel node biopsy, axillary sampling and axillary dissection in patients with breast cancer. AB - The aim of this study was to compare the incidence of lymphoedema after different treatments of the axilla in patients with breast cancer (BC). Medical records of 205 women (median age 61 years, range 26-72 years) who underwent curative surgery for primary BC were reviewed. According to the treatment of the axilla, the study population was divided into four age- and stage-matched groups of patients: Group A (N=54 patients), sentinel node biopsy (SLNB) alone; Group B (N=48 patients), SLNB followed by axillary node (AN) sampling using ultrasound scissors (harmonic scalpel); Group C (N=53 patients), AN dissection using ultrasound scissors; Group D (N=50 patients), traditional AN dissection. The median follow-up was 22 months (range 18-28 months). The intraoperative frozen section of SLNB (Groups A and B) showed 32 out of 102 (31.4%) patients with metastasis to AN, while final pathology showed AN metastases in 20, 17, 16 and 17 patients of groups A, B, C and D, respectively (p=NS). The sensitivity of SLNB alone was 80% and that of SLNB followed by AN sampling was 95% (p=NS). At follow-up patients with lymphoedema were 2 (3.7%), 2 (4.2%), 3 (5.6%) and 8 (16%) in groups A, B, C and D, respectively (p=NS). In conclusion, AN sampling is a sensitive and low morbidity procedure which, in conjunction with the use of harmonic scalpel, may reduce the onset of arm lymphoedema. PMID- 20023250 TI - Adrenomedullin concentration in second trimester amniotic fluid cannot be used as a predictor of preterm delivery. AB - BACKGROUND: Adrenomedullin, secreted by decidua and trophoblast cells, is considered to participate in regulating uterine and placental blood flow, leading to control of placental hormonal secretion. Furthermore, adrenomedullin has an antimicrobial activity. The objective of this study was to determine whether adrenomedullin concentrations in midtrimester amniotic fluid can be used as a predictor of preterm delivery. PATIENTS AND METHODS: Amniotic fluid samples were collected in a retrospective cross-matched study that included 362 women with singleton pregnancies who presented for genetic amniocentesis. Adrenomedullin concentrations were determined by ELISA in amniotic fluid taken from women with spontaneous preterm delivery (n=41) and maternal age-matched controls who had normal pregnancy at term (n=41). RESULTS: No difference was found in adrenomedullin concentrations between women with spontaneous preterm delivery (median: 1.33 ng/ml, range: 0.36-8.53 ng/ml) and controls (median: 1.32 ng/ml, range: 0.33-4.07 ng/ml), nor between a subset of cases of preterm premature rupture of membranes (n=19) and their controls (n=19). CONCLUSION: Adrenomedullin concentration in amniotic fluid cannot serve as a predictor of preterm delivery. PMID- 20023251 TI - Amebic liver abscesses: a new epidemiological trend in a non-endemic area? AB - BACKGROUND: Amoebic liver abscess (ALA) is the most common extraintestinal complication of colonic amebiasis. In recent decades its incidence in developed European countries has significantly increased because of travel and immigration of individuals from highly endemic areas. We report our 29-year experience in echo-guided percutaneous needle/catheter drainage (EPND/EPCD) of ALA. PATIENTS AND METHODS: From May 1979 to November 2007, 68 ALA corresponding to 56 patients were diagnosed at our Department. All patients were treated with a metronidazole plus EPND/EPCD approach. RESULTS: The majority of the cases did not need more than two echo-guided punctures. Two patients, both male immigrants (HIV negative), had unmodified lesions after two EPNDs: catheter drainage was performed. A quick worsening of their clinical conditions and onset of neurological symptoms occurred; in both patients, computed tomography (CT) revealed a brain abscess. Intravenous medical therapy was started, but both died 4 and 3 days, respectively, after the onset of neurological symptoms (overall mortality rate: 3.57%). CONCLUSION: The unfavorable outcome of two cases is a rare example of failure of percutaneous therapy of ALA. Mortality is a possible event even in a non-endemic area such as Italy. More observational data are needed to confirm the possibility of a new epidemiological trend. PMID- 20023253 TI - Challenges and opportunities for medical directors in pathology and laboratory medicine: standardization, integration, and innovation. AB - Because the practice of pathology and laboratory medicine evolves rapidly, laboratory medical directors must constantly introduce new tests and services and continue to provide consistent, reliable results for existing tests. Innovations in laboratory medicine are frequently introduced, and the number of commercial vendors of test kits and reagents increases yearly. These innovations, however, may pose barriers to standardization and integration of laboratories and to interpretation of results generated by different laboratories. We propose a practical framework for medical directors to address the seemingly contradictory challenges of standardizing and integrating while simultaneously providing the flexibility to introduce innovations. We recommend initiating standardization first, then integration, while maintaining flexibility for innovation. As organizations strive to create effective processes to enhance value, the role of the laboratory medical director will become critical in resolving the natural tension between standardization/integration and innovation in laboratory medicine and pathology. PMID- 20023254 TI - The effect of renal dysfunction on BNP, NT-proBNP, and their ratio. AB - We examined the effect of renal dysfunction on B-natriuretic peptide (BNP), N terminal (NT)-proBNP, and their molar ratio at varying severities of cardiac function in 94 Thai patients with chest pain (52 men; 32 women), also measuring creatinine and left ventricular ejection fraction (LVEF). Renal function was classified into 5 stages by estimated glomerular filtration rate. The molar NT proBNP/BNP ratio was calculated. Cardiac status was classified by LVEF (normal, >50%; moderate, 35%-50%; severe, <35%). BNP, NT-proBNP, and their ratio corresponded to renal disease stage exponential (0.51, 1.05, and 0.54, respectively; correlation coefficients, >or=0.95). BNP and the ratio are affected less than NT-proBNP by renal dysfunction, starting in stage III; NT-proBNP expresses effects starting in stage II. NT-proBNP is more sensitive than BNP to renal disease stage. For log of geometric means vs stage of renal disease, the BNP slopes and correlation coefficients vary considerably (slopes, 0.036-0.531; r(2), 0.017-0.99). The NT-proBNP slopes and regression coefficients vary considerably (slopes, 0.18-0.71; r(2), 0.33-0.99). For the ratio, the slopes show low variation (0.148-0.337), r(2) greater than 0.96, women differing from men (P = .012). The effect of renal disease differs by gender. BNP and NT-proBNP increase by stage III for women but not for men. One must consider renal function, gender, and LVEF when using BNP or NT-proBNP as cardiac biomarkers. The ratio of the 2 peptides is the most consistent marker across LVEFs. PMID- 20023255 TI - Application of the Toyota Production System improves core laboratory operations. AB - To meet the increased clinical demands of our hospital expansion, improve quality, and reduce costs, our tertiary care, pediatric core laboratory used the Toyota Production System lean processing to reorganize our 24-hour, 7 d/wk core laboratory. A 4-month, consultant-driven process removed waste, led to a physical reset of the space to match the work flow, and developed a work cell for our random access analyzers. In addition, visual controls, single piece flow, standard work, and "5S" were instituted. The new design met our goals as reflected by achieving and maintaining improved turnaround time (TAT; mean for creatinine reduced from 54 to 23 minutes) with increased testing volume (20%), monetary savings (4 full-time equivalents), decreased variability in TAT, and better space utilization (25% gain). The project had the unanticipated consequence of eliminating STAT testing because our in-laboratory TAT for routine testing was less than our prior STAT turnaround goal. The viability of this approach is demonstrated by sustained gains and further PDCA (Plan, Do, Check, Act) improvements during the 4 years after completion of the project. PMID- 20023256 TI - Cytoplasmic expression of nucleophosmin accurately predicts mutation in the nucleophosmin gene in patients with acute myeloid leukemia and normal karyotype. AB - Mutations in the nucleophosmin (NPM1) exon 12 resulting in delocalization of NPM1 into the cytoplasm occur in 50% to 60% of acute myeloid leukemia cases with a normal karyotype (AML-NK). As recent studies suggest such patients have a favorable prognosis and there are discordant reports of the immunohistochemical detection of cytoplasmic NPM1 (NPMc+) for predicting NPM1 gene mutations, we correlated the immunohistochemical detection of NPMc+, NPM1 gene mutations, and prognosis in 57 cases of AML-NK. All 31 NPMc+ cases (54% of total) had NPM1 mutations, but none of the 26 nucleus-restricted (NPMc-) cases (46% of total) had NPM1 mutations (P < .0001). NPM1 mutations were correlated with FLT3-internal tandem duplication (ITD) (P = .0062), absence of CD34 (P = .0001), and absence of CD7 (P = .041). There was a favorable survival outcome in AML-NK cases that were NPM1 mutated and FLT3-ITD nonmutated. Our data confirm that cytoplasmic NPM1 immunoreactivity predicts NPM1 mutations and warrants inclusion in the routine diagnostic and prognostic workup of AML. PMID- 20023257 TI - Expression profiling of transcription factors in B- or T-acute lymphoblastic leukemia/lymphoma and burkitt lymphoma: usefulness of PAX5 immunostaining as pan Pre-B-cell marker. AB - The optimal use of transcription factors to determine B-lineage specificity in B acute lymphoblastic leukemia/lymphoma (B-ALL) has not been fully investigated. We undertook an extensive immunohistochemical study of a panel of B-cell transcription factors in B- and T-ALL and Burkitt lymphoma to evaluate those with the best specificity and sensitivity. Tissue microarrays were constructed from 34 B-ALL, 19 T-ALL, and 30 Burkitt lymphoma samples. All 34 (100%) cases of B-ALL expressed PAX5; 32 (94%), BOB.1; 33 (97%), PU.1; 29 (85%), CD79a; 27 (79%), CD22; 2 (6%), CD20; 9 (26%), OCT-2; and 3 (9%), MUM1. Burkitt lymphoma cases were positive for PAX5 (30/30 [100%]), BOB.1 (27/30 [90%]), PU.1 (23/30 [77%]), CD79a (29/30 [97%]), CD22 (14/30 [47%]), CD20 (30/30 [100%]), OCT-2 (23/30 [77%]), and MUM1 (5/30 [17%]). T-ALLs were only positive for PU.1 (15/19 [79%]) and BOB.1 (12/19 [63%]). PAX5 demonstrated better specificity for B-lineage determination than BOB.1 and PU.1 and better sensitivity than CD79a, CD22, and CD20. These findings suggest that PAX5 has the greatest diagnostic usefulness and lineage determination in B-ALL, especially in cases with an inadequate specimen for flow cytometric analysis. PMID- 20023258 TI - Extranodal follicular dendritic cell sarcoma of the pharyngeal region: a potential diagnostic pitfall, with literature review. AB - Extranodal follicular dendritic cell sarcoma (FDCS) of the pharyngeal region is a rare malignant tumor recognized in recent years, with approximately 37 cases so far reported in the literature. It is often not considered at the initial evaluation and may be misdiagnosed in a small biopsy specimen. We report 4 cases of extranodal FDCS, 2 cases in the nasopharynx that were diagnosed as undifferentiated carcinomas because they were characterized by syncytial epithelial cells with sheet or nest-like distribution and 2 cases in the tonsil and soft palate that were characterized by vaguely concentric whorls consisting of spindle to ovoid cells. The latter case was diagnosed as ectopic meningioma. The analysis of all cases from the literature and ours shows that 58% (21/36) of the cases are misdiagnosed initially, often as undifferentiated carcinoma or meningioma, which the differential diagnoses should be mostly focused on. With a median follow-up of 27 months, the recurrence, metastasis, and mortality rates are 23%, 21%, and 3%, respectively, suggesting that extranodal FDCS of the pharyngeal region remains a low-grade sarcoma. Radical surgery is recommended, whereas there is no evidence to support adjuvant therapy. PMID- 20023259 TI - Characteristics of cutaneous marginal zone lymphomas with marked plasmacytic differentiation and a T cell-rich background. AB - Primary cutaneous marginal zone lymphoma (MZL) is a common B-cell lymphoma of skin and is characterized by an infiltrate of neoplastic marginal zone B cells typically within the marginal zones of reactive lymphoid follicles and the interfollicular region. However, in our experience, many cases have underemphasized features such as marked plasmacytic differentiation and/or a prominent T-cell component, which may obscure the neoplastic B cells and lead to misdiagnosis. We wanted to draw attention to these features and have studied 15 cases of MZL with marked plasmacytic differentiation, 10 of which had numerous T cells, some with cytologic atypia, and few B cells in the interfollicular region. Plasma cells were monotypic in all cases by in situ hybridization. By polymerase chain reaction, 6 of 8 T cell-rich cases had an IGH gene rearrangement, and none were clonal for T-cell receptor gene. We discuss the terminology, morphologic features, molecular profile, behavior, and differential diagnosis of cutaneous MZL. PMID- 20023260 TI - Aberrant nuclear p53 expression predicts hemizygous 17p (TP53) deletion in chronic lymphocytic leukemia. AB - Hemizygous TP53 gene deletion is the most important adverse risk factor in chronic lymphocytic leukemia (CLL), but its relationship with p53 protein expression is unclear. We investigated 110 CLL cases and correlated nuclear p53 protein immunoreactivity with TP53 gene deletion status and other CLL-associated genetic risk factors. Fluorescence in situ hybridization detected hemizygous TP53 deletions in 15 cases (13.6%), whereas immunohistochemical analysis detected nuclear p53 protein expression in 14 (12.7%). All cases expressing nuclear p53 protein had hemizygous TP53 deletions. Hemizygous TP53 gene deletion and p53 protein expression were strongly correlated (P < .001). There was no association between p53 expression and del(13q), del(11q) or trisomy 12 in CLL cases. Our data indicate that nuclear p53 protein expression, detected by a widely available immunohistochemical method, is strongly associated with TP53 deletion and that p53 immunohistochemical analysis may be adopted as a rapid, robust diagnostic tool for risk stratification of CLL. PMID- 20023261 TI - Therapy-related acute lymphoblastic leukemia without 11q23 abnormality: report of six cases and a literature review. AB - Therapy-related acute lymphoblastic leukemia (t-ALL) is a rare secondary leukemia following chemotherapy and/or radiotherapy for primary malignancies. Chromosomal 11q23 abnormality, frequently detected in therapy-related acute myeloid leukemia, is the most common cytogenetic alteration in t-ALL. However, t-ALL cases without 11q23 abnormality have been rarely described. We describe 6 adults with secondary t-ALL without 11q23 abnormalities following various treatment regimens for primary malignancies. We also reviewed 48 t-ALL cases, with complete chromosomal karyotyping, reported in the literature from 1992 to 2007. In the 48 cases, an 11q23 abnormality involving the MLL gene locus was the predominant chromosomal aberration (32 [67%]), followed by t(9;22) (6 [13%]) and a normal karyotype (4 [8%]). Compared with t-ALL cases with an 11q23 abnormality, cases without an 11q23 abnormality had a relatively longer latency period (median, 36 vs 19 months) and a different primary malignancy spectrum. No major difference was observed between groups in regard to age, sex, or receipt of a topoisomerase II inhibitor. The t(8;14)(q11.2;q32), a rare, nonrandom, balanced chromosomal translocation differing from the more common translocation involving c-MYC on chromosome 8q24, was seen in 1 adult t-ALL case, which may suggest another possible pathogenesis of this disease. PMID- 20023262 TI - Simultaneous cytomorphologic and multiparametric flow cytometric analysis on lymph node samples is faster than and as valid as histopathologic study to diagnose most non-Hodgkin lymphomas. AB - We evaluated the validity and accuracy of cytomorphology and multiparametric flow cytometry (C-FCM) in diagnosing oncohematologic disease in 223 consecutive lymph node biopsy specimens from patients with lymphadenopathy, from 2004 to 2007. C FCM and histopathologic studies were interpreted independently by hematologists and pathologists, respectively. C-FCM detected neoplastic disorders in 133 samples (59.6%): 92 non-Hodgkin lymphomas (NHLs; 41.3%), 21 Hodgkin lymphomas (HLs; 9.4%), 19 malignant nonhematologic neoplasms (8.5%), and 1 multiple myeloma (0.4%). Sensitivity and specificity were 87.25% and 95.95%, respectively. Positive predictive value and negative predictive value (NPV) were 97.74% and 78.89%, respectively. Sensitivity and NPV were 94.79% and 96.81% upon excluding HL and malignant nonhematologic neoplasms from the analysis. Of the 92 NHLs, 89 (97%) were categorized according to the 2001 World Health Organization classification of hematolymphoid neoplasms with a concordance of 87%. The C-FCM study was significantly faster than the histopathologic study. C-FCM has high sensitivity and specificity, allowing for a valid and reliable diagnosis, especially in NHLs and enabling their subclassification. C-FCM is faster than the histopathologic examination, allowing for therapeutic decisions to be made quickly. However, in the samples in which C-FCM cannot establish a diagnosis, histopathologic results are needed. PMID- 20023263 TI - Diagnosis of myelodysplastic syndrome among a cohort of 119 patients with fanconi anemia: morphologic and cytogenetic characteristics. AB - Predisposition to myelodysplastic syndrome (MDS) and acute leukemia is a hallmark of Fanconi anemia (FA). Morphologic criteria for MDS in FA are not well established, nor is the significance of clonal chromosomal abnormalities. We reviewed bone marrow samples of 119 FA patients: 23 had MDS, with the most common subtype refractory cytopenia with multilineage dysplasia. The presence of MDS was highly correlated with the presence of clonal abnormalities. Neutrophil dysplasia and increased blasts were always associated with the presence of a clone, in contrast with dyserythropoiesis. The most frequent clones had gains of 1q and 3q and/or loss of 7. Karyotype complexity also correlated with MDS. One third of patients with 3q as a sole abnormality had no MDS; patients with 3q and an additional abnormality all had MDS. The data provide a rationale for integrating cytogenetic findings with independently evaluated morphologic findings for monitoring bone marrow status in FA. PMID- 20023264 TI - Frequency of bacille Calmette-Guerin (BCG) and Mycobacterium tuberculosis in tissue biopsy specimens of children vaccinated with BCG. AB - Vaccination of all newborns with bacille Calmette-Guerin (BCG) vaccine is a standard practice in developing countries. Disseminated mycobacterial infection in an immunocompromised child can be caused by BCG and other mycobacteria. A total of 21 patients with a histopathologic diagnosis of mycobacterial infection were studied in a period of 4 years. DNA was extracted from formalin-fixed, paraffin-embedded tissues. Real-time polymerase chain reaction was performed to determine the mycobacterial species. The overall sensitivity of the assay was 71.5%. The prevalence rates of BCG, Mycobacterium tuberculosis, and other mycobacteria in the positive results were 80% (12/15), 13% (2/15), and 7% (1/15), respectively. PMID- 20023265 TI - Comparison of two rapid assays for Clostridium difficile Common antigen and a C difficile toxin A/B assay with the cell culture neutralization assay. AB - We compared 3 rapid assays for Clostridium difficile with a cell culture cytotoxicity neutralization assay (CCNA). Of 600 stool samples, 46 were positive for toxigenic C difficile. Both rapid common antigen assays were highly sensitive (91.3%-100%) and, therefore, were appropriate screening tests. The rapid toxin assay had poor sensitivity (61%) but excellent specificity (99.3%). Testing stools for glutamate dehydrogenase (step 1) and those positive with a rapid toxin assay (step 2) would correctly classify 81% of submitted specimens within 2 hours, including during periods of limited staffing (evenings, nights, and weekends). CCNA could then be used as a third step to test rapid toxin-negative samples, thereby providing a final result for the remaining 19% of samples by 48 to 72 hours. The use of rapid assays as outlined could enhance timely diagnosis of C difficile. PMID- 20023266 TI - Comparison of DiaSorin and Bio-Rad test kits for the detection of hepatitis B virus total core and surface antibodies on the Bio-Rad Evolis. AB - The new MONOLISA Bio-Rad kits were compared to DiaSorin kits for the detection of hepatitis B virus (HBV) total core (HBcTAb) and surface (HBsAb) antibodies on the Bio-Rad Evolis immunoanalyzer. The resolved sensitivities, specificities, positive and negative predictive values, and overall accuracy were 91%, 100%, 100%, 96%, and 97%, respectively, for HBcTAb and 99%, 95%, 96%, 99%, and 97%, respectively, for HBsAb. Whereas accuracy and reagent cost were comparable between the kits, Bio-Rad kits required less specimen volume and less instrument processing time to results than the DiaSorin kits. PMID- 20023267 TI - Diagnostic usefulness of fluorescent cytogenetics in differentiating chromophobe renal cell carcinoma from renal oncocytoma: a validation study combining metaphase and interphase analyses. AB - We investigated the usefulness of interphase fluorescence in situ hybridization (FISH) analysis to differentiate between 11 chromophobe renal carcinomas and 12 renal oncocytomas, showing different clinical outcomes, when compared with conventional metaphase cytogenetics by karyotyping. Karyotypically, 3 chromophobe renal cell carcinomas showed losses of chromosomes, 3 were polyploid, 1 was normal, and 4 failed to grow. Of 12 oncocytomas, 5 showed a normal numeric karyotype and 6 additional structural rearrangements. FISH on chromophobe renal cell carcinomas showed a high percentage of cases (10/11 [91%]) with multiple numeric losses among chromosomes 1, 2, 6, 10, and 17; this interphase pattern was observed irrespective of the 3 different metaphase karyotypes. Of 12 oncocytomas, 11 (92%) revealed a normal numeric chromosomal status showing at least 2 chromosomes without aneusomy by interphase FISH. The study demonstrates that indeed FISH performed on formalin-fixed, paraffin-embedded tissue can provide clinically useful information more reliably than karyotyping of most of these tumors. PMID- 20023268 TI - Non-viral-related pathologic findings in liver needle biopsy specimens from patients with chronic viral hepatitis. AB - Liver biopsy is a vital part of chronic viral hepatitis management. Pathologists should view these biopsies as screening tools for other liver diseases. We describe our experience in a 6-year period and discuss the pathologist's role. Liver biopsies of 1,842 patients with hepatitis B or C, for the 2001-2007 period at the University Health Network, Toronto, Canada, were reviewed; 410 other diagnoses were documented in 377 patients (20.5%; mean age, 25.4 years; range, 15 80 years). These diagnoses included 58 hepatocellular carcinomas and 16 dysplastic nodules, which are recognized complications of chronic viral hepatitis. The remaining findings included the following: steatosis/steatohepatitis, 251; hemosiderosis, 62; granulomatous disease, 7; drug induced hepatitis, 4; primary biliary cirrhosis, 3; Wilson disease, 2; metastases, 2; and cholangiocarcinoma, atypical lymphoid proliferation, alpha(1) antitrypsin deficiency, cystic fibrosis, and schistosomiasis, 1 each. Liver biopsies in patients with viral hepatitis revealed other processes with the potential to modify disease progression and/or the management strategy in 20.5% of patients. PMID- 20023269 TI - Primary oncocytic adenocarcinomas of the lung: a clinicopathologic, immunohistochemical, and molecular biologic analysis of 16 cases. AB - Sixteen cases of primary oncocytic adenocarcinomas of the lung are reported. The patients were 11 women and 5 men between the ages of 47 and 81 years (median, 75 years) with symptoms of cough, chest pain, and shortness of breath. Surgical staging disclosed 14 patients (88%) with stage I disease, 1 (6%) with stage II, and 1 (6%) with stage III. Histologically, all the cases displayed prominent oncocytic features with conventional growth patterns, including acinar, papillary, and bronchioloalveolar. Immunohistochemically, the tumors displayed positive staining for keratin 7, thyroid transcription factor-1, and mitochondrial antibody. Molecular studies showed 3 (20%) of 15 tumors with EGFR mutations and 3 additional cases with KRAS mutations. Clinical follow-up of at least 24 months was obtained in all patients and showed that 5 patients had recurrences, 2 patients died of tumor, and 2 other patients died of unrelated conditions. These cases represent an unusual variant of pulmonary adenocarcinoma. PMID- 20023270 TI - BRAF mutation status in gastrointestinal stromal tumors. AB - Gastrointestinal stromal tumors (GISTs) are mesenchymal tumors characterized by mutations of KIT or PDGFRA. The objectives of this study were to evaluate BRAF mutations in GISTs and then to correlate BRAF mutational status in the tumor with clinical parameters, with B-raf expression, and with activation of some cellular pathways. BRAF mutation was screened in 321 GISTs with 70 wild-type GISTs. BRAF V600E was detected in 9 (13%) of 70 wild-type GISTs. No mutations were detected in GISTs bearing KIT or PDGFRA mutations. BRAF V600E detection in the tumor does not induce a higher expression of the B-raf protein or the preferential activation of the p42/44 mitogen-activated protein kinase (MAPK) signaling pathway compared with GISTs without the BRAF mutation. In comparison with the GIST group with KIT or PDGFRA mutation or the wild-type GIST group without BRAF mutation, the wild-type GIST group with a BRAF mutation is not different in terms of B-raf expression or the p44/42 MAPK- or AKT-activated signaling pathway. PMID- 20023271 TI - Detection of c-KIT and PDGFRA gene mutations in gastrointestinal stromal tumors: comparison of DHPLC and DNA sequencing methods using a single population-based cohort. AB - Mutational analysis of c-KIT or PDGFRA has become an important laboratory assay for patients with gastrointestinal stromal tumors (GISTs) because the results are useful in predicting the responsiveness to imatinib. To assess the diagnostic usefulness of denaturing high-pressure liquid chromatography (DHPLC) in this setting, we performed DHPLC and DNA sequencing to study exons 9, 11, 13, and 17 of c-KIT and exons 12 and 18 of PDGFRA in 54 consecutive cases of GIST collected from a single population. Most (40/54 [74%]) carried c-KIT mutations, and 7 (13%) carried PDGFRA mutations. These results were similar to those described in the literature. It is important to note that DHPLC was found to be highly sensitive, detecting all of the mutations in these 6 exons that were identified by DNA sequencing. Our data suggest that DHPLC is a cost-effective, rapid, and sensitive test for screening for mutations of c-KIT and PDGFRA in GISTs. PMID- 20023272 TI - Experience with voice recognition in surgical pathology at a large academic multi institutional center. AB - There are few reports of institutional use of voice recognition technology in clinical practice. We describe our experience with voice recognition-integrated synoptic-like dictation, associating templates with key spoken phrases, that we have used in gross examination of common specimens and as a major component of our workflow since 2001. The primary application is VoiceOver Enterprise (Voicebrook, Lake Success, NY), which uses Dragon NaturallySpeaking Medical Edition (Nuance Communications, Burlington, MA) as its speech engine. This integrates with the anatomic pathology laboratory information system (APLIS) and other applications, such as Microsoft Office (Microsoft, Redmond, WA). The largest user group, pathology assistants, mainly dictates biopsy reports, numbering approximately 210,000 specimens since 2001. The technology has been useful in our anatomic pathology workflow and provided a good return on investment, including marked improvements in turnaround time, results standardization, error reduction, and cost savings. The most helpful features of the software are templating, the seamless integration with APLIS, and the voice command creation tools. PMID- 20023273 TI - Thrombin generation and plasma dilution. PMID- 20023274 TI - A survey of family members' satisfaction with the services provided by hospice palliative care volunteers. AB - A total of 22 family members, whose deceased loved ones had used the services of a hospice palliative care volunteer, responded to a brief survey designed to assess the importance of the different kinds of support offered to them (family members) by the volunteer, their impressions of the volunteers' personal qualities/characteristics, their general experiences with the volunteer, and their overall satisfaction with the volunteer services. The kind of support that received the highest importance rating from family members was the opportunity to take a much-needed break from the demands of caring for their loved one, closely followed by emotional support, the volunteer spending time with them, and the volunteer providing them with information. Family members rated volunteers highly on a list of qualities/characteristics that exemplify individuals who are effective in this role. In all, 85% of the family members felt that their volunteer was well trained and 95% did not feel that their or their loved one's privacy had been invaded by having a volunteer. Overall, family members were very satisfied with the volunteer support they received. Some limitations of the study are discussed. PMID- 20023275 TI - Hemodynamic analysis of arterial blood flow in the coiled umbilical cord. AB - The most significant anatomical structure of the umbilical cord is its level of coiling. The coiled geometry of the umbilical cord largely affects umbilical blood flow that is vital for fetus's well-being and normal development. In this study, we developed a computational model of steady blood flow through the coiled structure of an umbilical artery. The results showed that the driving pressure for a given blood flow rate is increasing as the number of coils in cord structure increases. The driving gradient pressures also vary with the pitch that dictates the coils' spreading. The coiled structure is resulting in interwoven streamlines along the helix and wall shear stresses (WSS) with significant spatial gradients along the cross-sectional perimeter anywhere within the helical coil. These gradients may have an adverse effect on the development of the fetus cardiovascular system in cases with over coiling (OC) or under coiling (UC) characteristics. The number of coils does not affect the distribution and levels of WSS. However, when the coils are more spread (eg, larger pitch number), the maximal WSS is significantly smaller. Cases with twisted and OC cords seem to yield very large values and gradients of WSS, which may place the fetus into high risk of abnormal development. PMID- 20023276 TI - Equine disease surveillance, July to September 2009. PMID- 20023277 TI - Third eyelid resection as a treatment for suspected squamous cell carcinoma in 24 horses. AB - Between October 2000 and January 2007, 24 horses were presented with suspected squamous cell carcinoma (SCC) of the third eyelid. The hospital's medical records were analysed retrospectively to gain data about the cases, and telephone follow up was obtained from the owners and referring veterinary surgeons. The resected third eyelid was submitted for histological examination in 21 cases; in the other three cases the tissue was not submitted at the owners' request, for economic reasons. SCC was confirmed in 16 of these 21 cases, three cases were diagnosed histologically as lymphoid hyperplasia, one as a mast cell tumour and one as a sebaceous gland adenocarcinoma. Long-term follow up over a median period of 41 months revealed no recurrence of SCC or associated problems; six of the 16 confirmed SCC cases were euthanased for unrelated reasons during the follow-up period. PMID- 20023278 TI - Effect of anthelmintics on reproductive performance and first-lactation culling rate in Holstein heifers. AB - Female Holstein calves were treated with ivermectin from birth to first oestrus to study the effect of parasitic burden and anthelmintic treatment on reproductive and productive performance. First oestrus, age at first service and age at calving were advanced by 30, 70 and 110 days, respectively (P<0.05), in ivermectin-treated animals compared with controls. No significant differences were observed in the conception rate, the number of services and the characteristics of the newborn calves and any problems at calving between the two groups. Daily milk yield, fat content in milk during first lactation, and the concentrations of growth hormone, insulin-like growth factor type 1, insulin and prolactin in serum were similar in both groups of cows. Culling during the first lactation was more common in untreated (47 per cent) than in treated (11 per cent) cows (P<0.05). PMID- 20023279 TI - Simultaneous analysis of the nasal shedding kinetics of field and vaccine strains of Bordetella bronchiseptica. AB - Groups of four two-week-old puppies were administered serial dilutions of an intranasal vaccine containing live Bordetella bronchiseptica and canine parainfluenza virus vaccine and housed individually in isolator cages. Three vaccinated groups and one unvaccinated control group were exposed to virulent B bronchiseptica four weeks after vaccination and evaluated. Nasal swabs for bacterial culture and sera for agglutination tests were taken from all the dogs every week for four weeks. The bacteria isolated were identified by growth on specific agar and by specific PCR to distinguish between vaccine and challenge strains. The vaccine strain persisted in the nasal cavity after vaccination but no adverse reactions were observed. Serum agglutination titres were raised in the vaccinated dogs at challenge. Vaccine strains were not isolated after the challenge from most of the vaccinated dogs. The challenge strain was shed in the dogs vaccinated with the lowest dose (10(6.0) cfu/dose) for two to three weeks but the other vaccinated groups (10(7.0) and 10(8.0) cfu/dose) shed the challenge strain transiently or not at all. Only the group vaccinated with 10(6.0) cfu/dose exhibited clinical signs after challenge. PMID- 20023280 TI - Isolation of bluetongue virus serotypes 15 and 21 in West Bengal, India. PMID- 20023281 TI - Seroprevalence of H5 avian influenza virus in birds in the United Arab Emirates. PMID- 20023282 TI - Prevalence of eight bovine viruses in sika deer (Cervus nippon yesoensis) in Japan. PMID- 20023283 TI - Contagious bovine pleuropneumonia--a reminder. PMID- 20023284 TI - Urinary capillariosis in a cat in the UK. PMID- 20023285 TI - Therapy for horses with chronic progressive lymphoedema. PMID- 20023286 TI - Support for professional colleagues. PMID- 20023287 TI - Growth retardation, impaired triacylglycerol catabolism, hepatic steatosis, and lethal skin barrier defect in mice lacking comparative gene identification-58 (CGI-58). AB - Comparative gene identification-58 (CGI-58), also designated as alpha/beta hydrolase domain containing-5 (ABHD-5), is a lipid droplet-associated protein that activates adipose triglyceride lipase (ATGL) and acylates lysophosphatidic acid. Activation of ATGL initiates the hydrolytic catabolism of cellular triacylglycerol (TG) stores to glycerol and nonesterified fatty acids. Mutations in both ATGL and CGI-58 cause "neutral lipid storage disease" characterized by massive accumulation of TG in various tissues. The analysis of CGI-58-deficient (Cgi-58(-/-)) mice, presented in this study, reveals a dual function of CGI-58 in lipid metabolism. First, systemic TG accumulation and severe hepatic steatosis in newborn Cgi-58(-/-) mice establish a limiting role for CGI-58 in ATGL-mediated TG hydrolysis and supply of nonesterified fatty acids as energy substrate. Second, a severe skin permeability barrier defect uncovers an essential ATGL-independent role of CGI-58 in skin lipid metabolism. The neonatal lethal skin barrier defect is linked to an impaired hydrolysis of epidermal TG. As a consequence, sequestration of fatty acids in TG prevents the synthesis of acylceramides, which are essential lipid precursors for the formation of a functional skin permeability barrier. This mechanism may also underlie the pathogenesis of ichthyosis in neutral lipid storage disease patients lacking functional CGI-58. PMID- 20023288 TI - In vivo labelling of resting monocytes in the reticuloendothelial system with fluorescent iron oxide nanoparticles prior to injury reveals that they are mobilized to infarcted myocardium. AB - AIMS: To evaluate the feasibility of loading resting monocytes/macrophages by intravenous (i.v.) injection of fluorescent iron oxide nanoparticles prior to injury and tracking of these cells in the very same animal to myocardial infarction (MI) by magnetic resonance imaging (MRI) and optical imaging. METHODS AND RESULTS: Rats were injected with fluorescent iron oxide nanoparticles (10 mg/kg) (n = 15) prior to injury. After disappearance of the nanoparticles from the blood, MI was induced. Monocytes/macrophages were then tracked in the very same animal by MRI and optical imaging. Control groups were (i) non-injected animals (n = 15), (ii) injected animals associated with a sham operation (n = 8), and (iii) animals treated with an anti-inflammatory agent (n = 6). The presence of iron-loaded cells can be detected by MRI in vivo in the infarcted myocardium. Here, we showed that the detection of inflammatory cells in vivo correlated well with ex vivo imaging (MRI and reflectance fluorescence) and histology. We also showed that the method is robust enough to depict changes in the inflammatory response. CONCLUSION: This study demonstrates that resting monocytes/macrophages can be loaded in vivo by a simple i.v. injection of fluorescent superparamagnetic iron oxide nanoparticles prior to injury and then tracked, in the same animal, in a model of ischaemia-reperfusion leading to myocardial infarct. Although previous studies of macrophages infiltration following MI have labelled the macrophages after injury, this study, for the first time, has pre-load the resting monocytes with fluorescent iron oxide nanoparticles. PMID- 20023289 TI - Non-invasive diagnosis of chylopericardium by cardiac magnetic resonance imaging. PMID- 20023290 TI - A cycle-based model to predict blastocyst transfer cancellation. AB - BACKGROUND: Extended culture for blastocyst transfer is thought to result in embryos with high implantation potential, enabling the transfer of fewer embryos and the reduction of multiple pregnancies after in vitro fertilization (IVF). However, one major drawback of extended culture is the risk of transfer cancellation if no blastocyst develops by Day 5, despite the observation of adequate early embryo development. We set out to develop a model to predict blastocyst transfer cancellation. METHODS: The model was built from 562 consecutive first IVF cycles and tested on an independent validation cohort. Multivariable logistic regression analysis was used to test the association of patient and cycle characteristics with the presence of cultured blastocysts on Day 5 and to create a nomogram. The model's performance was quantified by discrimination and calibration and clinical utility was evaluated for various thresholds. RESULTS: Fertilization technique, number and quality ratio of the Day 3 embryos and female age were independently associated with blastocyst development. The final prediction model showed an area under the curve (AUC) of 0.75 in the training set (95% confidence interval (CI): 0.73-0.77) and was well calibrated. The AUC for the validation data set was 0.80 (95% CI: 0.78-0.83) and calibration was acceptable. Using a decision threshold of 0.55, the model showed a negative predictive value of 80.5% and a false positive rate of 17%. CONCLUSION: A cycle-based model could prove clinically-relevant for reducing the incidence of cancelled Day 5 transfers. PMID- 20023291 TI - Transvaginal ultrasonography with bowel preparation is able to predict the number of lesions and rectosigmoid layers affected in cases of deep endometriosis, defining surgical strategy. AB - BACKGROUND: Successful surgical treatment of deep bowel endometriosis depends on obtaining detailed information about the lesions, prior to the procedure. The objective of this study was to determine the capability of transvaginal ultrasonography with bowel preparation (TVUS-BP) to predict the presence of one or more rectosigmoid nodules and the deepest bowel layer affected by the disease. METHODS: A prospective study of 194 patients with clinical and TVUS-BP suspected deep endometriosis submitted to videolaparoscopy. Image data were compared with surgical and histological results. RESULTS: With respect to bowel nodule detection and presence of at least two rectosigmoid lesions, TVUS-BP had a sensitivity of 97 and 81%, specificity 100 and 99%, positive predictive value (PPV) 100 and 93% and negative predictive value (NPV) 98 and 96%, respectively. Regarding diagnosis of infiltration of the submucosal/mucosal layer, TVUS-BP had a sensitivity of 83%, specificity 94%, PPV 77%, NPV 96%. CONCLUSIONS: These findings show that TVUS-BP is an adequate exam for evaluating the presence of one or more rectosigmoid nodules and the deepest layer affected in deep infiltrating bowel endometriosis, confirming the importance of this technique for defining the most appropriate surgical strategy to be implemented. PMID- 20023292 TI - Metformin influence on hormone levels at birth, in PCOS mothers and their newborns. AB - BACKGROUND: Polycystic ovary syndrome (PCOS) tends to run in families and excess intrauterine androgen exposure has been suggested as one possible cause of PCOS. We wanted to study the relationship between maternal and offspring sex hormone levels and the possible effects of metformin treatment in PCOS pregnancies. METHODS: We performed a post hoc analysis of a trial in which 40 pregnant women with PCOS were randomized in the first trimester, to use either metformin 850 mg twice daily or placebo until delivery. Maternal venous blood and umbilical arterial and venous blood samples were collected at delivery. Outcome measures were levels of androgens, estrogens and sex hormone binding globulin (SHBG). RESULTS: (i) In newborns, SHBG levels were higher in the metformin group. All other hormones, both in mothers and offspring, were unaffected by metformin treatment. (ii) Mothers, who gave birth to boys, had higher estrone and estradiol levels compared with those who gave birth to girls. (iii) Male newborns had higher levels of testosterone, androstanediol glucuronide and estradiol compared with females. (iv) Positive correlations were found between maternal and newborn levels of androstenedione, dihydrotestosterone and estradiol. CONCLUSIONS: Intrauterine metformin exposure seems to result in elevated SHBG levels in newborns. However, at birth, maternal and newborn androgen and estrogen levels are unaffected by metformin use in pregnancy. Although androgen and estrogen levels are higher in male newborns compared with females, maternal and newborn androgen and estrogen levels are highly correlated at birth. PMID- 20023293 TI - Pregnancy loss in lesbian and bisexual women: an online survey of experiences. AB - BACKGROUND: Although pregnancy loss is a distressing health event for many women, research typically equates women's experiences of pregnancy loss to 'married heterosexual women's experiences of pregnancy loss'. The objective of this study was to explore lesbian and bisexual women's experiences of miscarriage, stillbirth and neonatal death. METHODS: This study analysed predominantly qualitative online survey data from 60 non-heterosexual, mostly lesbian, women from the UK, USA, Canada and Australia. All but one of the pregnancies was planned. Most respondents had physically experienced one early miscarriage during their first pregnancy, although a third had experienced multiple losses. RESULTS: The analysis highlights three themes: processes and practices for conception; amplification of loss; and health care and heterosexism. Of the respondents, 84% conceived using donor sperm; most used various resources to plan conception and engaged in preconception health care. The experience of loss was amplified due to contextual factors and the investment respondents reported making in impending motherhood. Most felt that their loss(es) had made a 'significant'/'very significant' impact on their lives. Many respondents experienced health care during their loss. Although the majority rated the overall standard of care as 'good'/'very good'/'outstanding', a minority reported experiencing heterosexism from health professionals. CONCLUSIONS: The implications for policy and practice are outlined. The main limitation was that the inflexibility of the methodology did not allow the specificities of women's experiences to be probed further. It is suggested that both coupled and single non-heterosexual women should be made more visible in reproductive health and pregnancy loss research. PMID- 20023294 TI - Patients' conceptualization of cryopreserved embryos used in their fertility treatment. AB - BACKGROUND: Although IVF treatments using cryopreserved embryos are offered by most infertility centres, little is known about patients' opinions regarding treatment using these embryos. The objective of this study is to describe how patients think about (treatment with) cryopreserved embryos. METHODS: In-depth interviews based on the method of interpretative phenomenological analysis were conducted with patients undergoing IVF/ICSI treatment at the Department of Reproductive Medicine of the Ghent University Hospital between May and July 2006. Seven couples (one couple considered as one participant) and 11 female patients who attended the clinic without their partner were interviewed. RESULTS: Most participants knew little about medical-technical procedures involved in treatment with cryopreserved embryos. This was compensated by a high confidence in the medical team. However, seven of the eighteen participants thought the quality of cryopreserved embryos diminished during the storage period. A lack of knowledge about medical-technical procedures was compensated by metaphors related to other domains of experience, especially kitchen metaphors: this might add to the belief that frozen embryos have an expiry date. However, none of the patients who considered treatment with cryopreserved embryos as less effective ever thought of refusing this type of treatment or discussed this with medical staff. In addition, patients rarely discussed the moral status of their cryopreserved embryos. CONCLUSIONS: The beliefs of infertile patients about (the effectiveness of treatment with) cryopreserved embryos encompass misconceptions, and doubts and fears which may influence their decision-making but which are seldom discussed with the medical staff. PMID- 20023295 TI - Expression of eicosanoid biosynthetic and catabolic enzymes in peritoneal endometriosis. AB - BACKGROUND: Increased peritoneal eicosanoid concentrations have been reported in endometriosis patients and might be important in disease-associated pain and inflammation. Here, we evaluated the expression of key biosynthetic and catabolic enzymes involved in this abnormal eicosanoid production in peritoneal macrophages and endometriotic lesions. METHODS: Peritoneal macrophages, endometriotic lesions and matched eutopic endometrium were collected from endometriosis patients (n = 40). Peritoneal macrophages and eutopic endometrium samples were also collected from disease-free women (n = 25). Expression of type IIA secretory phospholipase A(2) (sPLA(2)-IIA), cyclooxygenase-2 (COX-2), microsomal prostaglandin E synthase 1 (mPGES-1), 15-hydroxyprostaglandin dehydrogenase (15-PGDH) and 5-lipoxygenase (5-LO) was quantified by real-time PCR, and these five key enzymes were localized by immunohistochemistry. RESULTS: sPLA(2)-IIA, COX-2 and mPGES-1 mRNA was significantly increased in peritoneal macrophages of endometriosis patients compared with controls (P = 0.006, P = 0.016 and P = 0.025, respectively). In endometriosis patients, sPLA(2)-IIA, mPGES-1 and 15-PGDH mRNA was significantly enhanced in peritoneal lesions compared with matched eutopic endometrium (P < 0.001, P < 0.001 and P = 0.005, respectively). In eutopic endometrium, a significant decrease in 15-PGDH mRNA was found in the endometriosis group compared with controls (P = 0.023). Finally, sPLA(2)-IIA, COX-2, mPGES-1 and 15 PGDH immunostaining was found mainly in endometrial glands, whereas 5-LO was distributed throughout the glands and stroma. CONCLUSIONS: Our study highlights an imbalance between eicosanoid biosynthesis and degradation in endometriosis patients. Both peritoneal macrophages and endometriotic lesions may be involved. Research into new molecules inhibiting biosynthetic enzymes (such as sPLA(2)-IIA and mPGES-1) and/or activating catabolic enzymes (such as 15-PGDH) may prove to be a major field of investigation in the development of targeted medical therapies. PMID- 20023296 TI - The experience of spontaneous pregnancy loss for infertile women who have conceived through assisted reproduction technology. AB - BACKGROUND: The purpose of this qualitative, phenomenological study was to explore the subjective experiences of infertile women who conceived through the use of assisted reproduction technology--ovarian stimulation, intrauterine insemination or IVF--only to lose their pregnancy at 2-16 weeks gestation. METHODS: Ten women participated in in-depth, tape-recorded interviews. After initial content analysis, a phenomenological analysis was undertaken to identify common themes in the participants' stories. RESULTS: Nine common themes were identified. These included: a sense of profound loss and grief; diminished control; a sense of shared loss with their partners; injustice or lack of fairness; ongoing reminders of the loss; social awkwardness; fear of re-investing in the treatment process or a subsequent pregnancy; the need to make sense of their experience; and feelings of personal responsibility for what had happened. CONCLUSIONS: Participants' experiences of pregnancy loss were embedded within their experiences of infertility and medical treatment, and shaped by their significant investment in having a child. A significant feature was their marked ambivalence regarding future reproductive options after their pregnancy loss, reflecting a unique overlay of prominent anxiety in their grief experience. PMID- 20023297 TI - Understanding the mechanisms of human tubal ectopic pregnancies: new evidence from knockout mouse models. AB - Ectopic pregnancy, a worldwide health problem, is potentially life-threatening and occurs in approximately 1.5-2% of all pregnancies in the western world; however, the precise mechanisms underlying the initiation and development of tubal ectopic pregnancy are unknown. Tubal abnormalities and dysfunction, such as altered contractility or abnormal ciliary activity, have been speculated to lead to tubal ectopic pregnancy. To elucidate the cellular and molecular mechanisms of the tubal transport process, several knockout (KO) mouse models have been developed. This review summarizes what has been learned from studies of the Fallopian tube in caspase-1, cannabinoid receptor and Dicer1 KO mice. Our understanding of the mechanisms which contribute to tubal ectopic pregnancy in humans may be enhanced through further study of these KO mouse models. PMID- 20023298 TI - Establishment of a protein frequency library and its application in the reliable identification of specific protein interaction partners. AB - The reliable identification of protein interaction partners and how such interactions change in response to physiological or pathological perturbations is a key goal in most areas of cell biology. Stable isotope labeling with amino acids in cell culture (SILAC)-based mass spectrometry has been shown to provide a powerful strategy for characterizing protein complexes and identifying specific interactions. Here, we show how SILAC can be combined with computational methods drawn from the business intelligence field for multidimensional data analysis to improve the discrimination between specific and nonspecific protein associations and to analyze dynamic protein complexes. A strategy is shown for developing a protein frequency library (PFL) that improves on previous use of static "bead proteomes." The PFL annotates the frequency of detection in co immunoprecipitation and pulldown experiments for all proteins in the human proteome. It can provide a flexible and objective filter for discriminating between contaminants and specifically bound proteins and can be used to normalize data values and facilitate comparisons between data obtained in separate experiments. The PFL is a dynamic tool that can be filtered for specific experimental parameters to generate a customized library. It will be continuously updated as data from each new experiment are added to the library, thereby progressively enhancing its utility. The application of the PFL to pulldown experiments is especially helpful in identifying either lower abundance or less tightly bound specific components of protein complexes that are otherwise lost among the large, nonspecific background. PMID- 20023299 TI - A targeted spatial-temporal proteomics approach implicates multiple cellular trafficking pathways in human cytomegalovirus virion maturation. AB - The assembly of infectious virus particles is a complex event. For human cytomegalovirus (HCMV) this process requires the coordinated expression and localization of at least 60 viral proteins that comprise the infectious virion. To gain insight into the mechanisms controlling this process, we identified protein binding partners for two viral proteins, pUL99 (also termed pp28) and pUL32 (pp150), which are essential for HCMV virion assembly. We utilized HCMV strains expressing pUL99 or pUL32 carboxyl-terminal green fluorescent protein fusion proteins from their native location in the HCMV genome. Based on the presence of ubiquitin in the pUL99 immunoisolation, we discovered that this viral protein colocalizes with components of the cellular endosomal sorting complex required for transport (ESCRT) pathway during the initial stages of virion assembly. We identified the nucleocapsid and a large number of tegument proteins as pUL32 binding partners, suggesting that events controlling trafficking of this viral protein in the cytoplasm regulate nucleocapsid/tegument maturation. The finding that pUL32, but not pUL99, associates with clathrin led to the discovery that the two viral proteins traffic via distinct pathways during the early stages of virion assembly. Additional investigation revealed that the majority of the major viral glycoprotein gB initially resides in a third compartment. Analysis of the trafficking of these three viral proteins throughout a time course of virion assembly allowed us to visualize their merger into a single large cytoplasmic structure during the late stages of viral assembly. We propose a model of HCMV virion maturation in which multiple components of the virion traffic independently of one another before merging. PMID- 20023300 TI - Modifications of protein environment of the [2Fe-2S] cluster of the bc1 complex: effects on the biophysical properties of the rieske iron-sulfur protein and on the kinetics of the complex. AB - The rate-determining step in the overall turnover of the bc(1) complex is electron transfer from ubiquinol to the Rieske iron-sulfur protein (ISP) at the Q(o)-site. Structures of the ISP from Rhodobacter sphaeroides show that serine 154 and tyrosine 156 form H-bonds to S-1 of the [2Fe-2S] cluster and to the sulfur atom of the cysteine liganding Fe-1 of the cluster, respectively. These are responsible in part for the high potential (E(m)(,7) approximately 300 mV) and low pK(a) (7.6) of the ISP, which determine the overall reaction rate of the bc(1) complex. We have made site-directed mutations at these residues, measured thermodynamic properties using protein film voltammetry to evaluate the E(m) and pK(a) values of ISPs, explored the local proton environment through two dimensional electron spin echo envelope modulation, and characterized function in strains S154T, S154C, S154A, Y156F, and Y156W. Alterations in reaction rate were investigated under conditions in which concentration of one substrate (ubiquinol or ISP(ox)) was saturating and the other was varied, allowing calculation of kinetic terms and relative affinities. These studies confirm that H-bonds to the cluster or its ligands are important determinants of the electrochemical characteristics of the ISP, likely through electron affinity of the interacting atom and the geometry of the H-bonding neighborhood. The calculated parameters were used in a detailed Marcus-Bronsted analysis of the dependence of rate on driving force and pH. The proton-first-then-electron model proposed accounts naturally for the effects of mutation on the overall reaction. PMID- 20023301 TI - Activation of the chloroplast monogalactosyldiacylglycerol synthase MGD1 by phosphatidic acid and phosphatidylglycerol. AB - One of the major characteristics of chloroplast membranes is their enrichment in galactoglycerolipids, monogalactosyldiacylglycerol (MGDG), and digalactosyldiacylglycerol (DGDG), whereas phospholipids are poorly represented, mainly as phosphatidylglycerol (PG). All these lipids are synthesized in the chloroplast envelope, but galactolipid synthesis is also partially dependent on phospholipid synthesis localized in non-plastidial membranes. MGDG synthesis was previously shown essential for chloroplast development. In this report, we analyze the regulation of MGDG synthesis by phosphatidic acid (PA), which is a general precursor in the synthesis of all glycerolipids and is also a signaling molecule in plants. We demonstrate that under physiological conditions, MGDG synthesis is not active when the MGDG synthase enzyme is supplied with its substrates only, i.e. diacylglycerol and UDP-gal. In contrast, PA activates the enzyme when supplied. This is shown in leaf homogenates, in the chloroplast envelope, as well as on the recombinant MGDG synthase, MGD1. PG can also activate the enzyme, but comparison of PA and PG effects on MGD1 activity indicates that PA and PG proceed through different mechanisms, which are further differentiated by enzymatic analysis of point-mutated recombinant MGD1s. Activation of MGD1 by PA and PG is proposed as an important mechanism coupling phospholipid and galactolipid syntheses in plants. PMID- 20023303 TI - Effect of nanostructured titanium on anodization growth of self-organized TiO2 nanotubes. AB - To understand the effect of substrate microstructure on the formation of TiO2 nanotubes, anodic oxidizations of commercially pure titanium subjected to surface mechanical attrition treatment (SMATed-Ti) and unSMATed-Ti in a glycol solution containing NH4F and small amounts of water were investigated. The SMATed-Ti exhibit a nanocrystallized surface layer containing a high density of grain boundaries compared with unSMATed-Ti. The anodization results show that the formed TiO2 nanotube layer on the SMATed-Ti is much thicker than that on the unSMATed-Ti. It is indicated that nanocrystallized Ti is propitious to the growth of TiO2 nanotubes; grain boundaries and dislocations play the leading role in accelerating the reaction rate and ion diffusion coefficient during anodization. In addition, nanocrystallization of Ti does not change surface morphologies and phase components of the TiO2 nanotubes. PMID- 20023302 TI - Gene deletion of 7,8-linoleate diol synthase of the rice blast fungus: studies on pathogenicity, stereochemistry, and oxygenation mechanisms. AB - Linoleate diol synthases (LDS) are heme enzymes, which oxygenate 18:2n-6 sequentially to (8R)-hydroperoxylinoleic acid ((8R)-HPODE) and to (5S,8R) dihydroxy-, (7S,8S)-dihydroxy-, or (8R,11S)-dihydroxylinoleic acids (DiHODE). The genome of the rice blast fungus, Magnaporthe oryzae, contains two genes with homology to LDS. M. oryzae oxidized 18:2n-6 to (8R)-HPODE and to (7S,8S)-DiHODE, (6S,8R)-DiHODE, and (8R,11S)-HODE. Small amounts of 10-hydroxy-(8E,12Z) octadecadienoic acid and traces of 5,8-DiHODE were also detected by liquid chromatography-mass spectrometry. The contribution of the 7,8-LDS gene to M. oryzae pathogenicity was evaluated by replacement of the catalytic domain with hygromycin and green fluorescent protein variant (SGFP) cassettes. This genetically modified strain Delta7,8-LDS infected rice leaves and roots and formed appressoria and conidia as the native fungus. The Delta7,8-LDS mutant had lost the capacity to biosynthesize all the metabolites except small amounts of 8 hydroxylinoleic acid. Studies with stereospecifically deuterated linoleic acids showed that (8R)-HPODE was formed by abstraction of the pro-S hydrogen at C-8 and antarafacial oxygenation, whereas (7S,8S)-DiHODE and (8R,11S)-DiHODE were formed from (8R)-HPODE by suprafacial hydrogen abstraction and oxygenation at C-7 and C 11, respectively. A mac1 suppressor mutant (Delta mac1 sum1-99) of M. oryzae, which shows cAMP-independent protein kinase A activity, oxygenated 18:2n-6 to increased amounts of (10R)-HPODE and (5S,8R)-DiHODE. Expression of the 7,8-LDS gene but not of the second homologue was detected in the suppressor mutant. This suggests that PKA-mediated signaling pathway regulates the dioxygenase and hydroperoxide isomerase activities of M. oryzae. PMID- 20023304 TI - A study of mesoporous silica-encapsulated gold nanorods as enhanced light scattering probes for cancer cell imaging. AB - Mesoporous encapsulation of gold nanorods (GNRs) in a silica shell of controllable thickness (4.5-25.5 nm) was realized through a single-step coating method without any intermediary coating. The dependence of localized surface plasmon resonance (LSPR) extinction spectra of the coated GNRs on the thickness of the silica shell was investigated with both simulation and experiments, which agreed well with each other. It was found that cetyltrimethyl ammonium bromide (CTAB) molecules, which act as surfactants for the GNRs and dissociate in the solution, greatly affect the silica coating. Mesoporous silica-encapsulated GNRs were also shown to be highly biocompatible and stable in bio-environments. Based on LSPR enhanced scattering, mesoporous silica-encapsulated GNRs were utilized for dark field scattering imaging of cancer cells. Biomolecule-conjugated mesoporous silica-encapsulated GNRs were specifically taken up by cancer cells in vitro, justifying their use as effective optical probes for early cancer diagnosis. Mesoporous silica can also be modified with functional groups and conjugated with certain biomolecules for specific labeling on mammalian cells as well as carrying drugs or biomolecules into biological cells. PMID- 20023305 TI - Compression of freestanding gold nanostructures: from stochastic yield to predictable flow. AB - Characterizing the mechanical response of isolated nanostructures is vitally important to fields such as microelectromechanical systems (MEMS) where the behaviour of nanoscale contacts can in large part determine system reliability and lifetime. To address this challenge directly, single crystal gold nanodots are compressed inside a high resolution scanning electron microscope (SEM) using a nanoindenter equipped with a flat punch tip. These structures load elastically, and then yield in a stochastic manner, at loads ranging from 16 to 110 microN, which is up to five times higher than the load necessary for flow after yield. Yielding is immediately followed by displacement bursts equivalent to 1-50% of the initial height, depending on the yield point. During the largest displacement bursts, strain energy within the structure is released while new surface area is created in the form of localized slip bands, which are evident in both the SEM movies and still-images. A first order estimate of the apparent energy release rate, in terms of fracture mechanics concepts, for bursts representing 5-50% of the structure's initial height is on the order of 10-100 J m(-2), which is approximately two orders of magnitude lower than bulk values. Once this initial strain burst during yielding has occurred, the structures flow in a ductile way. The implications of this behaviour, which is analogous to a brittle to ductile transition, are discussed with respect to mechanical reliability at the micro- and nanoscales. PMID- 20023306 TI - A study of the compartmentalization of core-shell nanoparticles through fluorescence energy transfer of dopants. AB - Hybrid organic-inorganic templates and core-shell nanoparticles were used as models to study the communication between fluorescent probes placed inside nanoparticles. The hybrid templates were prepared on the basis of a mixed surfactant system using octadecyltrimethoxysilane as a reactive amphiphile. The core-shell particles were obtained after coating of the templates with a siloxane shell, using the silanol groups on their surface. Atomic force microscopy imaging showed that the templates were made of a flexible material that flattened significantly after deposition on a substrate and evaporation of the solvent. Pyrene was sequestered by the templates in an aqueous suspension, which placed it in a nonpolar environment, as observed by its fluorescence response. Subsequently, double-doped templates were prepared by sequestering coumarin 153 (C153), with pyrene-doped hybrid templates. The communication between these probes was studied on the basis of their spectral properties, by means of fluorescence resonance energy transfer (FRET). Energy transfer between the dyes with efficiencies up to 55% was observed. Similarly, double-doped core-shell particles prepared on the basis of the hybrid templates were doped with this pair of dyes. Despite the presence of the shell, which was intended to increment the average separation between the probes, interaction of the dyes was observed, although with lower efficiencies. A similar study was performed with C153 and 4 (dicyanomethylene)-2-methyl-6-p-(dimethylamino)styryl-4H-pyran (DCM). FRET studies indicated that the probes were placed in proximity to each other. We confirmed these observations by means of fluorescence lifetime measurements, which showed a decrease in the lifetime of the donor upon addition of the acceptor. PMID- 20023307 TI - The synthesis of single PdAu bimetallic nanowire: feasibility study for hydrogen sensing. AB - Single PdAu bimetallic nanowires have been synthesized via the sequential processes of electrochemical deposition and dielectrophoresis (DEP). In the first step, Pd/Au grains on predefined Au electrodes were grown by electrochemical deposition and could assist effectively the formation of a single PdAu nanowire with a good directionality by the subsequent DEP process. The synthesized PdAu nanowires have an average of approximately 10 at.% of Pd, a good resistance of a few hundred Omega, diameters of 300 nm on average and lengths of up to 15 microm. Based on the single PdAu nanowire, hydrogen detection was demonstrated in the range from 100 to 2500 ppm. PMID- 20023308 TI - Branched nanotrees with immobilized acetylcholine esterase for nanobiosensor applications. AB - A novel lab-on-a-chip nanotree enzyme reactor is demonstrated for the detection of acetylcholine. The reactors are intended for use in the RISFET (regional ion sensitive field effect transistor) nanosensor, and are constructed from gold tipped branched nanorod structures grown on SiN(x)-covered wafers. Two different reactors are shown: one with simple, one-dimensional nanorods and one with branched nanorod structures (nanotrees). Significantly higher enzymatic activity is found for the nanotree reactors than for the nanorod reactors, most likely due to the increased gold surface area and thereby higher enzyme binding capacity. A theoretical calculation is included to show how the enzyme kinetics and hence the sensitivity can be influenced and increased by the control of electrical fields in relation to the active sites of enzymes in an electronic biosensor. The possible effects of electrical fields employed in the RISFET on the function of acetylcholine esterase is investigated using quantum chemical methods, which show that the small electric field strengths used are unlikely to affect enzyme kinetics. Acetylcholine esterase activity is determined using choline oxidase and peroxidase by measuring the amount of choline formed using the chemiluminescent luminol reaction. PMID- 20023309 TI - The rapid growth of 3 microm long titania nanotubes by anodization of titanium in a neutral electrochemical bath. AB - The length of titania nanotubes formed by anodization of 0.1 mm thick titanium foil was found to be a strong function of the pH of the electrolyte. The longest nanotubes were formed by using an electrolyte consisting of 1 M Na(2)SO(4) plus 5 wt% NH(4)F with pH 7. At this pH, after 30 min of anodization, 3 microm length nanotubular titania arrays with top diameters of approximately 50 nm and bottom diameters of 100 nm were produced. No acid was added to this electrolyte. The formation of titania nanotubes in neutral pH systems was therefore successful due to the excess NH(4)F in the electrolyte which increases the chemical dissolution process at the metal/oxide interface. Since the pH of the electrolyte at the top part of the nanotubes is kept very high, the dissolution of the nanotubes at the surface is minimal. However, the amount is adequate to remove the initial barrier layer, forming a rather well-defined nanoporous structure. All anodized foils were weakly crystalline and the transformation to anatase phase was achieved by heat treatment at temperatures from 200 to 500 degrees C for 1 h in air. Annealing at temperatures above 500 degrees C induce rutile phase formation and annealing at higher temperatures accelerates the diffusion of Ti(4+) leading to excessive growth and the nanotubular structure diminishes. PMID- 20023310 TI - Tight-binding calculation of optical gain in tensile strained [001]-Ge/SiGe quantum wells. AB - It is known that under a tensile strain of about 2% of the lattice constant, the energy of the bottom conduction state of bulk Ge at the Gamma point falls below the minimum at the L point, leading to a direct gap material. In this paper we investigate how the same condition is realized in tensile strained Ge quantum wells. By means of a tight-binding sp(3)d(5)s(*) model, we study tensile strained Ge/Si(0.2)Ge(0.8) multiple quantum well (MQW) heterostructures grown on a relaxed SiGeSn alloy buffer along the [001] direction. We focus on values of the strain fields at the crossover between the indirect and direct gap regime of the MQWs, and calculate band edge alignments, electronic band structures, and density of states. We also provide a numerical evaluation of the MQW material gain spectra for TE and TM polarization under realistic carrier injection levels, taking into account the leakages related to the occupation of the electronic states at the L point. The analysis of the different orbital contributions to the near-gap states of the complete structure allows us to give a clear interpretation of the numerical results for the strain-dependent TM/TE gain ratio. Our calculations demonstrate the effectiveness of the structures under consideration for light amplification. PMID- 20023311 TI - The fabrication of highly ordered and visible-light-responsive Fe-C-N-codoped TiO2 nanotubes. AB - A one-step method for the fabrication of Fe-C-N-codoped TiO2 nanotubes by electrochemical anodization is reported. The proposed method is both simple and efficient. The prepared samples were annealed at 550 degrees C for 3 h. The resulting nanotubes were characterized by SEM, XRD, XPS, EDX and UV-vis spectrophotometer. The results showed that the average tube diameter of the nanotubes was 70 nm and wall thickness was 20 nm and the average tube length was 2.4 microm. The doped TiO2 nanotubes exhibited strong absorption in the visible light region. The maximum photocurrent efficiency measured under solar simulator was 2.7%. PMID- 20023312 TI - Synthesis of silicon nanotubes with cobalt silicide ends using anodized aluminum oxide template. AB - Silicon nanotubes (SiNTs) are compatible with Si-based semiconductor technology. In particular, the small diameters and controllable structure of such nanotubes are remaining challenges. Here we describe a method to fabricate SiNTs intrinsically connected with cobalt silicide ends based on highly ordered anodic aluminum oxide (AAO) templates. Size and growth direction of the SiNTs can be well controlled via the templates. The growth of SiNTs is catalyzed by the Co nanoparticles reduced on the pore walls of the AAO after annealing, with a controllable thickness at a given growth temperature and time. Simultaneously, cobalt silicide forms on the bottom side of the SiNTs. PMID- 20023313 TI - A highly sensitive humidity sensor based on a nanofibrous membrane coated quartz crystal microbalance. AB - A novel humidity sensor was fabricated by electrospinning deposition of nanofibrous polyelectrolyte membranes as sensitive coatings on a quartz crystal microbalance (QCM). The results of sensing experiments indicated that the response of the sensors increased by more than two orders of magnitude with increasing relative humidity (RH) from 6 to 95% at room temperature, exhibiting high sensitivity, and that, in the range of 20-95% RH, the Log(Deltaf) showed good linearity. The sensitivity of fibrous composite polyacrylic acid (PAA)/poly(vinyl alcohol) (PVA) membranes was two times higher than that of the corresponding flat films at 95% RH. Compared with fibrous PAA/PVA membranes, the nanofibrous PAA membranes exhibited remarkably enhanced humidity sensitivity due to their high PAA content and large specific surface area caused by the formation of ultrathin nanowebs among electrospun fibers. Additionally, the resultant sensors exhibited a good reversible behavior and good long term stability. PMID- 20023314 TI - Intracellular ion monitoring using a gold-core polymer-shell nanosensor architecture. AB - In this study, we describe the design of new ratiometric fluorescent nanosensors, whose architecture is based on a gold core surrounded by a poly(vinyl alcohol) polyacetal shell. To the gold core, indicator dyes and reference dyes are attached via a cysteine linker. This nanosensor architecture is flexible with regards to the number and types of fluorophore linkages possible. The robust poly(vinyl alcohol)-polyacetal shell protects the fluorophores linked to the core from non-specific interactions with intracellular proteins. The nanosensors developed in this way are biocompatible and can be easily incorporated into mammalian cells without the use of transfection agents. Here, we show the application of these nanosensors for intracellular pH and sodium ion measurements. PMID- 20023315 TI - The role of metal film electron density in a surface plasmon polariton assisted light emitter. AB - A novel and simple Sm(3+)-doped-polymer/Ag/ SiO(2) trilayer structure was fabricated to investigate the impact of surface plasmon polaritons (SPPs) on the emission quantum efficiency of the dipole emitter Sm(3+). Photoluminescence (PL) enhancement factors for samples with different Ag thicknesses (30-70 nm) were found to be different when the thickness of the Sm(3+)-doped-polymer film was kept constant (220 nm). Theoretical analysis suggests that the electron density of the metal film plays an important role in the SPP assisted PL enhancement process, and this was confirmed in a Sm(3+)-doped-polymer/Ag/Au/ SiO(2) multilayer structure. PMID- 20023316 TI - Photoluminescence of self-assembled InAs quantum dots embedded in photonic crystal nanocavities with shifted air holes. AB - This study investigates the photoluminescence for self-assembled InAs quantum dots embedded in photonic crystal nanocavities as two of the air holes nearest the H1 cavity were shifted. A rapid decrease of resonant wavelength and quality factor for the cavity modes, in which the electric field patterns extended in the shifting direction, were found as the shift increased from 0.2 to 0.4 lattice constants. This phenomenon is interpreted as being caused by the formation of two point defects between the nearest and second nearest air holes. PMID- 20023317 TI - Molecular-receptor-specific, non-toxic, near-infrared-emitting Au cluster-protein nanoconjugates for targeted cancer imaging. AB - Molecular-receptor-targeted imaging of folate receptor positive oral carcinoma cells using folic-acid-conjugated fluorescent Au(25) nanoclusters (Au NCs) is reported. Highly fluorescent Au(25) clusters were synthesized by controlled reduction of Au(+) ions, stabilized in bovine serum albumin (BSA), using a green chemical reducing agent, ascorbic acid (vitamin-C). For targeted-imaging-based detection of cancer cells, the clusters were conjugated with folic acid (FA) through amide linkage with the BSA shell. The bioconjugated clusters show excellent stability over a wide range of pH from 4 to 14 and fluorescence efficiency of approximately 5.7% at pH 7.4 in phosphate buffer saline (PBS), indicating effective protection of nanoclusters by serum albumin during the bioconjugation reaction and cell-cluster interaction. The nanoclusters were characterized for their physico-chemical properties, toxicity and cancer targeting efficacy in vitro. X-ray photoelectron spectroscopy (XPS) suggests binding energies correlating to metal Au 4f(7/2) approximately 83.97 eV and Au 4f(5/2) approximately 87.768 eV. Transmission electron microscopy and atomic force microscopy revealed the formation of individual nanoclusters of size approximately 1 nm and protein cluster aggregates of size approximately 8 nm. Photoluminescence studies show bright fluorescence with peak maximum at approximately 674 nm with the spectral profile covering the near-infrared (NIR) region, making it possible to image clusters at the 700-800 nm emission window where the tissue absorption of light is minimum. The cell viability and reactive oxygen toxicity studies indicate the non-toxic nature of the Au clusters up to relatively higher concentrations of 500 microg ml(-1). Receptor-targeted cancer detection using Au clusters is demonstrated on FR(+ve) oral squamous cell carcinoma (KB) and breast adenocarcinoma cell MCF-7, where the FA-conjugated Au(25) clusters were found internalized in significantly higher concentrations compared to the negative control cell lines. This study demonstrates the potential of using non-toxic fluorescent Au nanoclusters for the targeted imaging of cancer. PMID- 20023318 TI - Enhanced thermal conductivities of nanofluids containing graphene oxide nanosheets. AB - Stable ethylene-glycol-based nanofluids containing graphene oxide nanosheets have been prepared. The measurements of thermal conductivity indicate that the nanofluids have substantially higher thermal conductivities than the base fluid. The thermal conductivity enhancement depends strongly on the volume fraction of graphene oxide nanosheets and increases with the increasing loading. When the nanosheet loading is 5.0 vol%, the enhancement ratio is up to 61.0%. The thermal conductivity of the fluids remains almost constant for seven days, indicating their high stability. The level of enhancement is independent of temperature in the measured temperature range. PMID- 20023319 TI - Dynamic [Cl(-)](i) measurement with chloride sensing quantum dots nanosensor in epithelial cells. AB - We have synthesized a chloride sensing quantum dots (QD) nanosensor, Cl-QD, for the dynamic measurements of chloride ion concentration in the millimolar range, a sensitivity that is applicable to most physiological intracellular chloride ion concentration ([Cl(-)](i)) measurements in epithelial cells. The Cl-QD is synthesized by conjugating an anion receptor, 1-(2-mercapto-ethyl)-3-phenyl thiourea (MEPTU) to a water soluble CdSe/ZnS QD at an emission wavelength of 620 nm. Upon binding of chloride ions to the Cl-QD, a photo-induced electron transfer mechanism caused the fluorescence of the QD to quench. This resulted in an inversely proportional relationship between the chloride ion concentration and the fluorescence intensity of the Cl-QD. We have utilized this Cl-QD to measure [Cl(-)](i) in T84 and CF-PAC cultured cells, with either the C1C-2 or CFTR chloride channels being manipulated by pharmacological chloride channel activators and inhibitors. Activations of C1C-2 and CFTR chloride channels in T84 by the respective lubiprostone and genistein caused predictive increases in the fluorescence of the Cl-QD, i.e., a decrease of [Cl(-)](i). Conversely, glibenclamide, a chloride channel inhibitor, applied to the CF-PAC cells caused a predictable decrease in the fluorescence of Cl-QD due to the increase of [Cl( )](i). These are the first data in using QD-based chloride ion sensors for dynamic measurements of intracellular chloride ion concentrations in epithelial cells. PMID- 20023320 TI - Adhesion mechanisms of nanoparticle silver to substrate materials: identification. AB - Nanoparticle silver (NPS) conductors are increasingly being investigated for printed electronics applications. However, the adhesion mechanism of the nanoparticle silver to substrate materials has not been identified yet. In particular, the adhesion of NPS to organic materials such as the widely used polyimide Kapton HN and Kapton FPC dry films is concerned with low adhesion strength because the processed polymer surface is chemically inert. Moreover, its adhesion to substrate materials such as benzocyclobutene (BCB), copper and aluminum was found to be very weak. Therefore, in this paper, the mechanisms of NPS adhesion to organic and inorganic materials are identified as the first step in improving NPS adhesion strength. Improving the adhesion strength of NPS will be the key issue for printed electronics applications. The adhesion of NPS to substrate materials was found to be mainly attributed to van der Waals forces based on particle adhesion mechanisms. This finding provides the initiative of developing an adhesion prediction model of NPS to substrate materials in order to provide guidelines for improving the NPS adhesion strength to the substrate materials used in printed electronics. PMID- 20023321 TI - Lattice-resolved frictional pattern probed by tailored carbon nanotubes. AB - In this study, we demonstrate a high-resolution friction profiling technique using synchronous atomic/lateral force microscopy (AFM/LFM). The atomic resolution is achieved by our special carbon nanotube (CNT) probes made via in situ tailoring and manipulation inside an ultra-high vacuum transmission electron microscope (UHV TEM). The frictional pattern mapped on graphite displays a periodic distribution similar to the atomic (0001)-oriented graphite lattice structure. Furthermore, the electrothermal process in the UHV TEM renders a graphite-capped CNT tip, which delivers the nanotribology study within two graphite layers by the LFM measurement on graphite. The synchronous AFM and LFM images can discern a spatial shift between the atomic points and local friction maxima. We further interpret this shift as caused by the lattice distortion, which in turn induces irreversible energy dissipation. We believe this is the origin of atomic friction on the sub-nanonewton scale. PMID- 20023322 TI - A microcantilever heater-thermometer with a thermal isolation layer for making thermal nanotopography measurements. AB - This paper presents a microcantilever having a microscale heater-thermometer fabricated from doped single crystal silicon that is mounted on a silicon nitride thermal isolation structure. The silicon nitride isolation structure is in turn connected to doped single crystal silicon legs. The cantilever fabrication, its characterization, and its application in thermal nanotopography measurements are presented in this work. The cantilever can reach temperatures over 600 degrees C with a heating power of 4 mW. The cantilever has a thermal resistance that exceeds 10(5) K W(-1) when away from a substrate. Making a contact-mode scan over a silicon calibration grating of height 20 nm, the cantilever has a topography reading sensitivity of 1.3 x 10(-4) nm(-1), and a topography reading resolution of about 7 pm Hz(-1/2). These performance characteristics compare extremely well to published ones for other kinds of cantilevers. PMID- 20023323 TI - A cradle-shaped gradient coil to expand the clear-bore width of an animal MRI scanner. AB - The never ending quest for higher magnetic field strengths in MRI and MRS has led to small and medium bore scanners at 9.4 T and above for both human and animal use; however, these bore diameters restrict the size of object that can be accommodated when using a conventional gradient coil. By replacing a cylindrical gradient-coil insert with a single-sided gradient coil, the scanner's functionality can be extended to include localized imaging of wider samples. As a prototype, a three-axis, cradle-shaped gradient coil was designed, fabricated and implemented in a 9.4 T animal MRI scanner. Since gradient fields are required only to be monotonic over the desired field of view, the cradle gradient coil was designed to produce high gradient efficiencies (up to 2.25 mT m(-1) A(-1) over a 5 cm imaging region) at the expense of gradient linearity. A dedicated three dimensional algorithm was developed to correct the resultant image distortion. Preliminary images of a grid phantom and a mouse demonstrated the fidelity of the algorithm in correcting image distortion of greater than 200%. Eddy currents were measured along each gradient axis. A large 65.2 (Hz mT(-1) m) B(0) eddy current was produced by the y-axis, suggesting potential limitations of single-sided gradient coils. PMID- 20023324 TI - A method for measuring the energy spectrum of coincidence events in positron emission tomography. AB - Positron emission tomography (PET) system energy response is typically characterized in singles detection mode, yet there are situations in which the energy spectrum of coincidence events might be different than the spectrum measured in singles mode. Examples include imaging with isotopes that emit a prompt gamma in coincidence with a positron emission, imaging with low activity in a LSO/LYSO-based cameras, in which the intrinsic activity is significant, and in high scatter situations where the two 511 keV photons have different scattering probabilities (i.e. off-center line source). The ability to accurately measure the energy spectrum of coincidence events could be used for validating simulation models, optimizing energy discriminator levels and examining scatter models and corrections. For many PET systems operating in coincidence mode, the only method available for estimating the energy spectrum is to step the lower and upper level discriminators (LLD and ULD). Simple measurement techniques such as using a narrow sliding energy window or stepping only the LLD will not yield a spectrum of coincidence events that is accurate for cases where there are different energy components contributing to the spectrum. In this work we propose a new method of measuring the energy spectrum of coincidence events in PET based on a linear combination of two sets of coincident count measurements: one made by stepping the LLD and one made by stepping the ULD. The method was tested using both Monte Carlo simulations of a Siemens microPET R4 camera and measured data acquired on a Siemens Inveon PET camera. The results show that our energy spectrum calculation method accurately measures the coincident energy spectra for cases including the beta/gamma spectrum of the (176)Lu intrinsic activity present in the LSO scintillator crystals, a (68)Ge source and an (124)I source (in which there are prompt gamma-rays emitted together with the positron). PMID- 20023325 TI - An anatomically shaped lower body model for CT scanning of cadaver femurs. AB - Bone specific, CT-based finite element (FE) analyses have great potential to accurately predict the fracture risk of deteriorated bones. However, it has been shown that differences exist between FE-models of femora scanned in a water basin or scanned in situ within the human body, as caused by differences in measured bone mineral densities (BMD). In this study we hypothesized that these differences can be reduced by re-creating the patient CT-conditions by using an anatomically shaped physical model of the lower body. BMD distributions were obtained from four different femora that were scanned under three conditions: (1) in situ within the cadaver body, (2) in a water basin and (3) in the body model. The BMD of the three scanning protocols were compared at two locations: proximally, in the trabecular bone of the femoral head, and in the cortical bone of the femoral shaft. Proximally, no significant differences in BMD were found between the in situ scans and the scans in the body model, whereas the densities from the water basin scans were on average 10.8% lower than in situ. In the femoral shaft the differences between the three scanning protocols were insignificant. In conclusion, the body model better approached the in situ situation than a water basin. Future studies can use this body model to mimic patient situations and to develop protocols to improve the performance of the FE models in actual patients. PMID- 20023326 TI - Hybrid plan verification for intensity-modulated radiation therapy (IMRT) using the 2D ionization chamber array I'mRT MatriXX--a feasibility study. AB - The 2D ionization chamber array I'mRT MatriXX (IBA, Schwarzenbruck, Germany) has been developed for absolute 2D dosimetry and verification of intensity-modulated radiation therapy (IMRT) for perpendicular beam incidence. The aim of this study is to evaluate the applicability of I'mRT MatriXX for oblique beam incidence and hybrid plan verification of IMRT with original gantry angles. For the assessment of angular dependence, open fields with gantry angles in steps of 10 degrees were calculated on a CT scan of I'mRT MatriXX. For hybrid plan verification, 17 clinical IMRT plans and one rotational plan were used. Calculations were performed with pencil beam (PB), collapsed cone (CC) and Monte Carlo (MC) methods, which had been previously validated. Measurements were conducted on an Elekta SynergyS linear accelerator. To assess the potential and limitations of the system, gamma evaluation was performed with different dose tolerances and distances to agreement. Hybrid plan verification passed the gamma test with 4% dose tolerance and 3 mm distance to agreement in all cases, in 82-88% of the cases for tolerances of 3%/3 mm, and in 59-76% of the cases if 3%/2 mm were used. Separate evaluation of the low dose and high dose regions showed that I'mRT MatriXX can be used for hybrid plan verification of IMRT plans within 3% dose tolerance and 3 mm distance to agreement with a relaxed dose tolerance of 4% in the low dose region outside the multileaf collimator (MLC). PMID- 20023327 TI - Optimal margin and edge-enhanced intensity maps in the presence of motion and uncertainty. AB - In radiation therapy, intensity maps involving margins have long been used to counteract the effects of dose blurring arising from motion. More recently, intensity maps with increased intensity near the edge of the tumour (edge enhancements) have been studied to evaluate their ability to offset similar effects that affect tumour coverage. In this paper, we present a mathematical methodology to derive margin and edge-enhanced intensity maps that aim to provide tumour coverage while delivering minimum total dose. We show that if the tumour is at most about twice as large as the standard deviation of the blurring distribution, the optimal intensity map is a pure scaling increase of the static intensity map without any margins or edge enhancements. Otherwise, if the tumour size is roughly twice (or more) the standard deviation of motion, then margins and edge enhancements are preferred, and we present formulae to calculate the exact dimensions of these intensity maps. Furthermore, we extend our analysis to include scenarios where the parameters of the motion distribution are not known with certainty, but rather can take any value in some range. In these cases, we derive a similar threshold to determine the structure of an optimal margin intensity map. PMID- 20023328 TI - Cerenkov radiation allows in vivo optical imaging of positron emitting radiotracers. AB - In this paper, we showed that Cerenkov radiation (CR) escaping from the surface of small living animals injected with (18)F-FDG can be detected with optical imaging techniques. (18)F decays by emitting positrons with a maximum energy of 0.635 MeV; such positrons, when travelling into tissues faster than the speed of light in the same medium, are responsible of CR emission. A detailed model of the CR spectrum considering the positron energy spectrum was developed in order to quantify the amount of light emission. The results presented in this work were obtained using a commercial optical imager equipped with charged coupled detectors (CCD). Our data open the door to optical imaging (OI) in vivo of the glucose metabolism, at least in pre-clinical research. We found that the heart and bladder can be clearly identified in the animal body reflecting the accumulation of the (18)F-FDG. Moreover, we describe two different methods based on the spectral analysis of the CR that can be used to estimate the depth of the source inside the animal. We conclude that (18)F-FDG can be employed as it is as a bimodal tracer for positron emission tomography (PET) and OI techniques. Our results are encouraging, suggesting that it could be possible to apply the proposed approach not only to beta(+) but also to pure beta(-) emitters. PMID- 20023329 TI - Enhancement of irradiation effects on cancer cells by cross-linked dextran-coated iron oxide (CLIO) nanoparticles. AB - We investigated iron oxide nanoparticles with two different surface modifications, dextran coating and cross-linked dextran coating, showing that their different internalization affects their capability to enhance radiation damage to cancer cells. The internalization was monitored with an ultrahigh resolution transmission x-ray microscope (TXM), indicating that the differences in the particle surface charge play an essential role and dominate the particle cell interaction. We found that dextran-coated iron oxide nanoparticles cannot be internalized by HeLa and EMT-6 cells without being functionalized with amino groups (the cross-linked dextran coating) that modify the surface potential from 18 mV to 13.4 mV. The amount of cross-linked dextran-coated iron oxide nanoparticles uptaken by cancer cells reached its maximum, 1.33 x 10(9) per HeLa cell, when the co-culture concentration was 40 microg Fe mL(-1) or more. Standard tests indicated that these internalized nanoparticles increased the damaging effects of x-ray irradiation, whereas they are by themselves biocompatible. These results could lead to interesting therapy applications; furthermore, iron oxide also produces high contrast for magnetic resonance imaging (MRI) in the diagnosis and therapy stages. PMID- 20023330 TI - X-ray beam filtration, dosimetry phantom size and CT patient dose conversion factors. AB - We examine how the choice of CT x-ray beam filtration and phantom size influences patient dose (D) to computed tomography dose index (CTDI) conversion factors (i.e. D/CTDI). The ratio of head to body phantom CTDI(w) for a defined scan technique is alpha, and the ratio of organ dose when the body filter is changed to the head filter is beta. CTDI and organ doses were obtained using the ImPACT CT patient dosimetry calculator, and values of alpha and beta were determined for 39 CT scanners. The average value of alpha for the 39 CT scanners covering a 20 year period was 1.99 +/- 0.23, but 30% of scanners had alpha values that differed by more than 10% from the average. For GE, the value of alpha has been approximately constant at approximately 2.0. Both Philips and Siemens show a definite upward trend from values well below 2.0 in the early 1990s to well over 2.0 for their latest models. The data for Toshiba show no overall trend with time with half the data points below 2.0 and the remainder above this value. The average value of beta was 1.09 +/- 0.25. All vendors showed a downward trend in the beta parameter, and where the most recent scanners from each vendor had a beta value close to unity. Our results show that average D/CTDI conversion factors for a body phantom/filter combination are typically double those appropriate for a head phantom/filter combination. PMID- 20023331 TI - Simulation study of spatial resolution and sensitivity for the tapered depth of interaction PET detectors for small animal imaging. AB - Improvements to current small animal PET scanners can be made by improving the sensitivity and the spatial resolution of the scanner. In the past, efforts have been made to minimize the crystal dimensions in the axial and transaxial directions to improve the spatial resolution and to increase the crystal length to improve the sensitivity of the scanner. We have designed tapered PET detectors with the purpose of reducing the gaps between detector modules and optimizing the sensitivity of a future-generation small animal PET scanner. In this work, we investigate spatial resolution and sensitivity of a scanner based on tapered detector elements using Monte Carlo simulations. For tapered detector elements more scintillation material is used per detector resulting in a higher sensitivity of the scanner. However, since the detector elements are not uniform in size, degradation in spatial resolution is also expected. To investigate characteristics of tapered PET detectors, the spatial resolution and sensitivity of a one-ring scanner were simulated for a system based on traditional cuboid detectors and a scanner based on tapered detectors. Additionally, the effect of depth of interaction (DOI) resolution on the spatial resolution for the traditional and tapered detectors was evaluated. All simulations were performed using the Monte Carlo simulation package GATE. Using the tapered arrays, a 64% improvement in the sensitivity across the field of view was found compared with traditional detectors for the same ring diameter. The level of DOI encoding was found to be the dominating factor in determining the radial spatial resolution and not the detector shape. For all levels of DOI encoding, no significant difference was found for the spatial resolution when comparing the tapered and the cuboid detectors. Detectors employing the tapered crystal design along with excellent DOI resolution will lead to PET scanners with higher sensitivity and uniform spatial resolution across the field of view. PMID- 20023332 TI - Local corticosteroid treatment for carpal tunnel syndrome: a 6-month clinical and electrophysiological follow-up study. AB - OBJECTIVE: To evaluate the clinical and electrophysiological effects of local depo-methylprednisolone injection in patients with carpal tunnel syndrome (CTS) over a 6-months period. METHODS: Twenty one patients, of whom 7 were lost for follow-up (mean age 57.9 +/- 8.4) with clinical and electrophysiological evidence of CTS were treated by injection of depo-methylprednisolone 40 mg proximal to the carpal tunnel. Severity of pain (VAS), rates of numbness/paresthesias and nocturnal awakening, motor and sensory nerve conduction studies were used as outcomes. All tests were performed before, 1, 3 and 6 months after the injection. RESULTS: Severity of pain was significantly reduced at all follow-up time points (p < 0.001). Prior to injection all patients complained of night pain and awakening. On the first, third and sixth months, 0(0%), 4 (29%) and 7 (50%) of the patients, respectively, had night awakening. All patients complained of numbness before the treatment. This symptom disappeared in 81% of the patients after one month and reappeared in all after three months. Significant improvement was shown in the mean distal motor latency (DML) of the median nerve: 5.2 +/- 0.9 msec. before, 4.6 +/- 0.6 msec. and 4.7 +/- 0.6 msec. 1 and 3 months after the injection, respectively (p < 0.05). Mean values of motor muscle potential amplitudes, sensory latency and sensory amplitude did not change significantly after the treatment. CONCLUSIONS: Local corticosteroid injection for the treatment of CTS provides significant symptom improvement for three months. No electrophysiological parameters were improved after injection, except the improvement in distal motor latency of the median nerve. PMID- 20023333 TI - Can workers with chronic back pain shift from pain elimination to function restore at work? Qualitative evaluation of an innovative work related multidisciplinary programme. AB - INTRODUCTION: Workers with chronic low back pain (LBP) mean a heavy human and social-economic burden. Their medical histories often include different treatments without attention to work-relatedness or communication with occupational health providers, leaving them passive and medicalized in (outpatient) health care. So we developed and implemented an innovative, patient activating alternative: the multidisciplinary outpatient care (MOC) programme, including work(place) intervention and graded activity. It aims at function restore (instead of pain elimination), return to work (RTW) and coordinated communication. OBJECTIVES: To qualitatively explore how patients and health care providers perceive the programme effectiveness and which factors influence its implementation. METHODS: In-depth, semi structured interview with patients and focus groups of health care providers are used, all recorded, transformed into verbatim transcript and analysed. RESULTS: This qualitative study shows that although patients' expectations were low at the start of the program, and despite long LBP histories, including many different therapies, (primarily) directed at pain reduction, the MOC programme was successful in changing patients' goal setting from pain oriented towards function restore and RTW. The programme was therefore perceived as applicable and effective. Patient compliance was influenced by barriers - despair, supervisory and subordinate resistance at work, waiting period, medicalisation in health care - and facilitators: disciplinary motivation, protocolled communication, information supply, tailor-made exercises. For some patients the barriers were too high. Several improvement suggestions were given. CONCLUSIONS: This qualitative study shows that generally, patients and professionals perceived the multidisciplinary outpatient care programme as applicable and effective. After incorporating improvement suggestions this program seems promising for further, broader application and hypothesis testing. For those, negatively evaluating the programme, alternatives should be explored. PMID- 20023334 TI - Effects of Knight-Taylor brace on balance performance in osteoporotic patients with vertebral compression fracture. AB - OBJECTIVE: To assess the changes in static and dynamic balance and movement strategies in patients with severe osteoporotic vertebral compression fracture while wearing and not wearing the Knight-Taylor (K-T) spinal brace. SUBJECTS: 47 patients with severe osteoporotic vertebral compression fracture, which was confirmed on radiographs and with bone density measurements obtained by dual energy X-ray absorption. INTERVENTION: Patients were randomly subjected to computerized dynamic posturography, which contained sensory organization tests, motor control balance test at 75% limit of stability (LOS) in 8 movement directions, and left/right rhythmic weight shift test (L/R RWS), while wearing and not wearing the K-T spinal brace, respectively. RESULTS: Patients wearing the spinal brace had significantly increased average stability, significantly increased average maximal stability under the swayed vision with fixed support surface condition and under the eye open with swayed support surface condition, significantly increased ankle strategy and decreased average velocity of COG target sway under the eye open with swayed support surface condition, significantly reduced the frequency of falls under the eye closed with swayed support surface condition and swayed vision with swayed support surface condition, and significantly decreased in the percentage of directional control with no difference of reaction time in the LOS test, and an increase in the on axis velocity in the L/R RWS test. CONCLUSIONS: The K-T spinal brace efficiently enables the subjects to maintain static and dynamic motor balance. Its use decreases the fall frequency but limits the directional control in severe osteoporotic patients with vertebral compression fracture. PMID- 20023335 TI - Comparison of effectiveness according to different approaches of epidural steroid injection in lumbosacral herniated disk and spinal stenosis. AB - PURPOSE: This study was to compare the effectiveness of the translaminar, caudal, and transforaminal technique with small and large volume of injectate in the treatment of lumbosacral herniation of intervetebral disc (HIVD) or spinal stenosis (SS). METHOD: Medical records reviewed retrospectively were of patients with radicular pain over 3 months and had diagnosed as HIVD or SS. Patients who underwent four weeks of treatments such as analgesics, anti-inflammatory drugs, or physical therapy for radicular pain with no improvement and afterwards received ESI were selected. Exclusion criteria was those who had suffered trauma, patients with other serious diseases, patients demonstrating adverse reactions to the medications, and patients with more than two levels of disease. The number of HIVD and SS group is 95 and 138, respectively. Visual Analog Scale (VAS) pain score, Patient Satisfaction Index (PSI), and Roland 5-point pain score were compared between different epidural approaches at pretreatment, 2 weeks, 1 month, and 2 months after treatment. RESULTS: Higher ratio of successful results was found in translaminar and transforaminal techniques than caudal technique in VAS in the HIVD group and in VAS and PSI in the SS group. Reduction of Roland score was maintained until 2 months in all techniques in HIVD and SS groups. In SS group, transforaminal groups showed more reduction of Roland score than caudal approach. No difference was found between small and large volume of transforaminal techniques. CONCLUSION: Translaminar and transforaminal approach were more effective than caudal approach in HIVD and SS groups. Especially, effectiveness of transforaminal approach was more prominent in SS group as compared with HIVD group. PMID- 20023336 TI - Sacroiliac joint pathologies in low back pain. AB - OBJECTIVE: The study describes the clinical spectrum of patients with low back pain due to sacroiliac joint (SIJ) involvement with the proposition of a diagnostic scheme. METHODS: In this retrospective review, 61 patients with SIJ pain (unilateral or bilateral) greater than six weeks duration were evaluated by pain history, clinical examination including SIJ provocative tests, laboratory investigations and skeletal imaging. RESULTS: Fifty two patients (M: F, 31:21) were diagnosed to have specific SIJ pathologies amongst 61 patients presenting between 2002 to 2004. Forty patients (65%) were diagnosed with rheumatic conditions - ankylosing spondylitis (AS) - 21, undifferentiated spondyloarthropathy (UspA) - 11, psoriatic arthropathy (PS) - 5, reactive arthropathy (ReA) - 1 and juvenile spondyloarthropathy (JS)-2. Non rheumatic conditions were involved in 12 patients (20%) - osteitis condensus ilii (OCI) - 4, osteomalacia - 2, tuberculosis - 2, pyogenic arthritis - 1, pregnancy related sacroiliac joint pain - 2 and malignancy in 1 patient. The diagnosis could not be confirmed in 9 patients (15%). CONCLUSIONS: Medical history, clinical examination including SIJ tests, plain radiography and laboratory investigations were helpful in diagnosing SIJ pathology in 39% cases (n = 24), 46% (n = 28) needed CT or MRI. A diagnostic scheme of dividing the SIJ pathologies into rheumatic and non rheumatic conditions was helpful in evaluating patients with suspected SIJ pathologies. PMID- 20023337 TI - Health education home-based program in females with fibromyalgia: a pilot study. AB - The objective of this study was to assess the long- and short-term effects of a multimodal program, specifically designed to be carried out by fibromyalgia syndrome (FMS) patients themselves, after a 12-week period of learning. Thirty female FMS patients volunteered for the study. The learning phase consisted on twelve weekly sessions of an hour, combining muscular resistance and flexibility exercises with techniques of breathing and relaxing, plus a half-hour incentive physiotherapy session. Flexibility, illness impact and pain were assessed at the beginning of the study, at the end of the learning phase and six months afterwards (follow-up). Twenty-one patients completed the programme. At the end of the learning phase, the improvement in the patients' physical condition was significant, as was the impact of the illness. Thirty-three percent of the patients continued training during the follow-up period and kept up the improvements achieved. The self-controlled multimodal programme was efficient in improving flexibility and reducing the impact of the illness in women fibromyalgia patients. However, adherence was poor when patients had to exercise on their own. PMID- 20023339 TI - Aerobic fitness and limiting factors of maximal performance in chronic low back pain patients. AB - BACKGROUND: Studies of measurement of maximal aerobic uptake in patients with chronic low back pain have shown inconsistent results and none has focused on clinical endpoints of ergometry tests. OBJECTIVE: To determine the level of cardiorespiratory fitness and to establish factors limiting the maximal effort during the ergometry. METHODS: Patients with chronic low back pain performed a graded maximal exercise test. Clinical endpoints of the test were determined and grouped as "maximal effort" and "symptom-limited effort" endpoints. RESULTS: 101 patients aged 29.8 +/- 7.5 achieved a VO2 max value of 30.0 (+/- 7.27) ml.kg( 1).min(-1). In men and women independently, a linear regression analysis demonstrated that VO2 max was inversely and weakly related to age. When compared to normative categories of VO2 max, men and women were situated in the fair category. Quadriceps/leg fatigue was reported by 47.5% of patients and was the most frequent limiting factor of the tests. "Symptom limited effort" endpoints were reported by 54.4% of the subjects. CONCLUSIONS: Clinical limiting factors of maximal exertion interfere the achievement of maximal oxygen uptake in patients with low back pain during ergometry. Further, there exists an association among the clinical endpoints of the tests and the VO2 value achieved. CLBP patients have a lower level of aerobic fitness than healthy controls. PMID- 20023338 TI - Long segment instrumentation of thoracolumbar burst fracture: fusion versus nonfusion. AB - OBJECTIVE: The treatment of thoracolumbar burst fracture is a controversial issue. Although spinal fusion has been a touchstone of spinal fixation, nonfusion technique have become raising its popularity recently. Some studies suggested that nonfusion had several advantages over fusion. The aim of this prospective study was to compare long segment posterior instrumentation with fusion versus long-segment posterior instrumentation without fusion. METHODS: For this purpose, 42 consecutive patients were assigned to two groups. Group 1 included 21 patients treated by long segment instrumentation with fusion (WF), whereas Group 2 included 21 patients treated by long segment instrumentation without fusion (WOF). Long segment instrumentation was hook fixation (claw hooks attached to second upper vertebra and infralaminar hooks attached to first upper vertebra) above and pedicle fixation (pedicle screws attached to first and second lower vertebrae) below the fractured vertebra. RESULTS: Measurements of local kyphosis, sagittal index and anterior vertebral height compression showed that both group had similar outcome at final follow-up. Moreover, there was no difference between the two groups according to low back outcome score. Also, implant failure rate (4.7%) was quite low in both groups. However, WF group had prolonged operative time, increased blood loss and donor site morbidity. CONCLUSIONS: Radiological and clinical parameters demonstrated that spinal fusion is not necessary in long segment posterior instrumentation for the management of thoracolumbar burst fractures. PMID- 20023340 TI - A case of brucellar spondylodiscitis involving the cervical spine. AB - We have presented a patient of 71-year-age with brucellar spondylodiscitis, involving the cervical spine, especially the C3-C4 segment. The patient had painful percussion of the cervical spine and passive mobilization of the neck, decreased range of motion, and cervical paravertebral tenderness; but no abnormalities observed on neurological examination. Wright agglutination test for brucella was positive at 1/320. Cervical localization for brucellar spondylodiscitis is an unusual case and should be detected and treated as early as possible. In endemic regions, spinal involvement of brucellosis should be considered in cases with fever, neck and low back pain. PMID- 20023341 TI - Adult case of acrodysostosis with severe neurologic involvement. AB - OBJECTIVE: Acrodysostosis is a rare syndrome characterized by peripheral dysostosis, nasal hypoplasia and frequently mental retardation. Only one adult case of acrodysostosis has been reported to have neurologic symptoms. We report one further adult case of acrodysostosis with severe neurologic findings including myelopathy and spastic paraparesis due to diffuse spinal stenosis and recurrent deep vein thrombosis possibly caused by neurologic deficits. RESULTS: We report a 43-year-old woman who had back and neck pain with weakness in the extremities of several years. 1~year before admission to our hospital, she had been treated with a missed diagnosis of sero (-) spondyloarthropathy but had not benefited. She became unable to walk, thereafter she underwent decompression surgery with a diagnosis of degenerative spinal stenosis. She presented at our outpatient department complaining of lowback pain and difficulty walking. She had marked facial and peripheral appearance of acrodysostosis. Spinal MRI revealed extensive spinal stenosis. A diagnosis was made through the genetic investigation, clinical and radiological findings. Spastic paraparesis were detected. There was widespread neuropathic pain. 15 days after admission, she developed swelling and redness of the left lower extremity and the venous doppler ultrasonography showed left acute and right past DVT. We treated DVT with anticoagulant therapy. Gabapentin and Baclofen were initiated for neuropathic pain and spasticity. A conventional rehabilitation program was performed. She left walking with a walker without pain and spasticity. CONCLUSIONS: We would like to remind physicians to be aware of peripheral malformations as signs of skeletal dysplasias and to consider acrodysostosis in the differential diagnosis. Although it is a rare condition, if diagnosed early, possible complications can be treated and outcomes may be improved. PMID- 20023342 TI - Trunk stabilization training: an evidence basis for the current state of affairs. PMID- 20023343 TI - Hamstring flexibility in young women following passive stretch and muscle energy technique. AB - BACKGROUND: Research is limited and controversial on the effectiveness of various stretching methods on muscle lengthening. OBJECTIVE: Studying the effectiveness of static stretch and muscle energy technique on the flexibility of hamstring muscles. METHODS: Thirty young females (20-25 yrs old) participated voluntarily in this study. Subjects were randomly assigned to two groups: static stretch group (controls; n=15) and muscle energy technique (MET) group (experimental; n=15). The control group underwent ten sessions of static stretches of hamstring muscle, and the experimental group was assigned to ten sessions of MET at 50% voluntary isometric contraction. Hamstring flexibility was evaluated by passive knee extension test at baseline and again following interventions. RESULTS: Both treatment methods were capable of significantly improving the shortness of hamstring muscles (P< 0.01); and no considerable difference was observed between the methods. CONCLUSION: Ten sessions of hamstring stretches in normal young women using either static stretch or MET had similar effect on restoring flexibility to hamstrings. PMID- 20023344 TI - Reliability of quality measures of movement in lumbar spine flexion-extension radiography. AB - BACKGROUND DATA: For center of rotation (COR) and centroid length variables in lumbar flexion-extension radiography, enough reliability information is not available. OBJECTIVE: The objective of this study was to determine the intra rater reliability of COR and centroid length of lumbar intervertebral movement. METHODS: A methodological research for reliability was designed. Fifteen males with chronic non specific low back pain aged between 22-43 years participated in this study. Study variables included COR of full, flexion and extension arc and centroid length. The proposed method from Putto was used for flexion-extension radiography. Five x-rays were taken in neutral, full flexion and extension, mid flexion and mid-extension positions. The variables were calculated using Computer Aided Radiographic Analysis of Spine (CARA) software after scanning. The intraclass correlation coefficient (ICC) and standard error of measurement (SEM) were utilized for relative and absolute reliability respectively. RESULTS: The results demonstrated high ICC values on x axis (range: 0.67-0.88) and y axis (range: 0.55-0.90) for full arc COR, high ICC values of flexion arc COR on x axis (range: 0.54-0.97) and y axis (range: 0.48-0.99). ICC values results of extension arc ranged between 0.19-0.60 for x axis and 0.16-0.83 for y axis. ICC values for centroid length ranged between 0.22-0.88. CONCLUSION: Results support previous studies which show very high reliability of full arc COR. In addition, flexion arc COR has desirable reliability and low measurement errors. But, extension arc COR and centroid length have moderate reliability and large measurement errors. PMID- 20023345 TI - Intra and inter-rater reliability of a goniometric lower trunk rotation measurement. AB - OBJECTIVES: Limited lower trunk rotation, which includes rotation of the lumbar spine, may hinder or even prevent functional activities. Currently, due to the lack of reliable, valid, and clinically useful tests, there is no standard objective measure of lower trunk rotation that can be easily performed in the clinic. The purpose of this study was to establish a standard protocol and to determine inter-rater and intra-rater reliability for a goniometric measurement developed to measure lower trunk rotation. METHODS: Lower trunk rotation was measured using a specific, goniometric method in 41 subjects. Each subject was measured 6 times by 2 different examiners for a total of 12 measurements. RESULTS: Pearson correlation coefficients indicate good intra-rater reliability ranging from 0.59 to 0.82 for right rotation (P< 0.001) and 0.76 to 0.82 for left rotation (P< 0.001), as well as good inter-rater reliability ranging from 0.62 to 0.83 with right rotation (P< 0.001) and 0.75 to 0.77 for left rotation (P< 0.001). CONCLUSION: This measure of trunk rotation may be useful for objectively documenting lower trunk rotation. PMID- 20023346 TI - Evaluation of trunk muscle activity in chronic low back pain patients and healthy individuals during holding loads. AB - OBJECTIVES: Low back pain after load-carrying is the most important disorder in musculoskeletal system and a cause of dysfunction and economic problems. Holding materials can disturb spinal stability; nevertheless, there are few researches about the pattern of trunk muscle recruitment in patients with chronic low back pain (CLBP) during load holding. METHODS: Ten female patients with CLBP and ten matched healthy subjects participated in this study. Normalized electromyography activation of trunk muscles during holding loads was analyzed. RESULTS: The low back pain group demonstrated significantly higher activation levels of the External oblique abdominis muscle during loading 12 kg in flexed trunk position and lower activation levels of the Internal oblique abdominis muscle during loading 6 and 12 kg in neutral trunk position than the control group. With the highest external load and trunk flexion, the electrical activity of back muscles increased significantly in both groups. With increasing load, the activation of Rectus abdominis muscle in patients with CLBP and the activation of Rectus abdominis and Internal oblique muscles were increased significantly in healthy subjects. CONCLUSION: Higher activation of global and lower activation of local abdominal muscles in patients with CLBP may represent that pain changes neuromuscular control systems. The increased activity of extensor muscles during trunk flexion is probably needed for stability and controlling of flexion. PMID- 20023347 TI - Computerized back postural assessment in physiotherapy practice: Intra-rater and inter-rater reliability of the MIDAS system. AB - BACKGROUND AND PURPOSE: Assessment of spinal posture during physiotherapy practice is routine, yet few objective measures exist to this end. The Middlesbrough Integrated Digital Assessment System (MIDAS) is a low cost portable system able to record 3D information on posture. The purpose of this study was to assess both the intra-rater and inter-rater reliability of the MIDAS system. METHODS: Twenty-five healthy subjects were recruited. A repeated measures design was used to record fifteen pre-palpated landmarks on the back of each subject. To limit the sources of variability, the principal researcher palpated the landmarks for each subject. Each of three raters took two measurements on each subject in a standardized upright posture. X (medio-lateral), Y (antero-posterior) and Z (height) landmark positions were recorded via a computer interface. RESULTS: Both intra-rater agreement (mean ICCs - rater 1 r=0.970, rater 2 r=0.965 and rater 3 r=0.965, p< 0.001) and inter-rater agreement (mean ICCs r=0.967, p< 0.001) was very high between repeated measures and between markers. Error values for the z axis (height) were the lowest. CONCLUSIONS: The MIDAS demonstrated both high inter-rater and intra-rater reliability and provides an objective method for the assessment of posture in physiotherapy practice. PMID- 20023348 TI - Efficacy of two multimodal treatments on physical strength of occupationally subgrouped male with low back pain. AB - OBJECTIVES: Some occupations are more prone to low back pain (LBP) due to their static work postures and work place design. Multidisciplinary pain programs have shown their effectiveness in the management of LBP in general population but which treatment and which segment of the population will be benefited more was not investigated yet. This study determines the effect of two treatment protocols on five occupationally subgrouped male LBP patients. METHODS: A total of 102 occupational male, 20-40 yrs of age, with sub-acute or chronic nonspecific LBP were randomized and treated either with conventional treatment a combination of two electrotherapy (ultrasound and short wave diathermy) and one exercise therapy (lumbar strengthening exercises) or dynamic muscular stabilization techniques (DMST) an active approach of stabilizing training. At the end of the treatment, subjects of both the groups were further stratified in five subgroups on the basis of their occupation. The pain was the primary outcome measure while physical strength [back pressure changes (BPC) and abdominal pressure changes (APC)] the secondary. RESULTS: The Pain, BPC and APC of all subgroups improved significantly (P < 0.01) in the both treatments but more in DMST. For each variable, improvement in subgroups differed within and between the treatments. Overall improvement in all assessed variables were evident on Desk workers followed by Shop keepers the most while BPC of Movement job, APC of Others and Pain of Sedentary and Shop keepers improved the least. CONCLUSIONS: Study concluded that for the management of occupational LBP, DMST is more effective than conventional treatment. The Pain of Sedentary and Shopkeepers and physical strength of Movement job and Others may need more clinical attention. Findings of this study may be helpful in the management of occupational LBP. PMID- 20023349 TI - Association between age, sex, BMI and CT-evaluated spinal degeneration features. AB - OBJECTIVE: The aim of our study was to evaluate the association between age, sex and body mass index (BMI) and lumbar spine degeneration features evaluated on computed tomography (CT) in a community-based sample. METHODS: This cross sectional study was an ancillary project to the Framingham Study. A sample of 3529 participants of the Framingham study aged 40-80 had a CT scan performed to assess aortic calcification. 187 individuals were randomly enrolled in this ancillary study. The prevalence of intervertebral disc narrowing, facet joint osteoarthritis (OA), spondylolysis, spondylolisthesis and spinal stenosis were evaluated. To evaluate the association between spinal degeneration features and age, sex and BMI we used chi2 test and logistic regression analyses. RESULTS: 104 men and 83 women, mean age 52.6 +/- 10.8 participated in the study. Statistically significant sexual dimorphism was found in prevalence of spondylolysis (p = 0.015) the male-to-female ratio was 3.3:1; and degenerative spondylolisthesis (p=0.008), the male-to-female ratio was 1:2.8. Prevalence of disc narrowing, facet joint OA, and degenerative spondylolisthesis showed a significant linear trend (p < 0.0001) of association with increasing age. Significantly higher prevalence of facet joint OA was found in the obese group OR (95%CI): 2.8 (1.1 7.2). CONCLUSIONS: Marked differences in the prevalence of spinal degeneration features occur in association with age, sex and obesity. Given the high prevalence of many of these degeneration features these simple demographic factors should be considered when interpreting imaging results reporting these features. PMID- 20023350 TI - The correlation between craniovertebral angle, backpack weights, and disability due to neck pain in adolescents. AB - PURPOSES: To investigate the responses of the craniovertebral (CV) angle to backpack loadings in adolescents with and without neck pain and to explore the relationships between CV angle, relative backpack weight, neck pain and disability. METHODS: A cross-sectional single-blinded study was conducted on 60 adolescents (30 neck pain and 30 non-neck pain) aged from 13 to 18 years old. The verbal analog scale (VAS) and Chinese version of Northwick Park Neck Pain Questionnaire (NPQ) were used to assess neck pain severity and disability respectively. CV angle was measured in neutral and with backpack loadings of 5% to 30% of subject's body weight by using the Head Posture Spinal Curvature Instrument (HPSCI). RESULTS: In both groups, CV angles gradually decreased with increment of backpack loadings and the amount of decreases became significant from 10% body weight onwards (P < 0.05). Although the changes of CV angles did not show any significant differences at any point of comparison between the groups, the neck pain group showed a clinically significant decrease of CV angle ( approximately 5 degrees ) at 10% relative loading whereas non-neck pain group did it at 15% relative loading. Change of CV angles did not show significant correlations with relative backpack weight, cervical pain and disability (P > 0.05). CONCLUSIONS: Our findings suggested a safety limit of 10% relative backpack load for adolescents. The results showed the tendency that the ability of maintaining good head posture in response to backpack loadings by non-neck pain subjects might be better than those with neck pain. PMID- 20023351 TI - Isometric endurance of the back extensors in school-aged adolescents with and without low back pain. AB - BACKGROUND AND OBJECTIVE: Studies on back extensor endurance in adolescents are scarce. This study sought to establish reference data and pattern of back extensor endurance in school-aged adolescents with and without low-back pain (LBP) from Nigeria. SUBJECTS AND METHODS: This study recruited 625 adolescents aged 11 to 19 years from eight randomly selected secondary schools. The modified Biering-Sorensen test of Static Muscular Endurance (BSME) was used to assess isometric endurance of the back extensors. Demographic and anthropometric data were collected. A modified LBP questionnaire was used to assess the presence of LBP. Descriptive and inferential analyses were used to analyze data. Significance was set at 0.05 alpha-level. RESULTS: The mean isometric holding time (IHT) of all the participants was 132.9 $?pm$ 65.6. Males had significantly higher significant (p=0.026) IHT than females. Adolescents without LBP had a higher significant IHT (p=0.042) than those with reported history of previous LBP and those with present LBP (p=0.000) respectively. Using percentile values, poor endurance was defined as IHT that is < 90.0 s and < 67 s for males and females respectively; medium endurance was defined as IHT that ranged between 90 and 193 s and 67 and 170 s for males and females respectively while good endurance was defined as IHT that is > 193 s and > 170 s for males and females respectively. IHT was significantly related to each of body mass index, hip circumference and waist-to-hip ratio (p < 0.05). CONCLUSION: Isometric back extensors endurance in Nigerian adolescents was comparable to the original Biering-Sorensen mean value. Majority of the participants had medium endurance performance with the back endurance pattern in the ratio 1:2:1. Male had higher isometric back extensors endurance than females. Decreased isometric back extensors endurance was associated with the presence of LBP in adolescents. PMID- 20023352 TI - Effects of body weight supported treadmill training on cardiac and pulmonary functions in the patients with incomplete spinal cord injury. AB - We aimed to investigate the effects of body weight supported treadmill training (BWSTT) on cardio-pulmonary functions and on depression in subjects with incomplete spinal cord injury (SCI). Eight subjects (2 female, 6 male) with incomplete SCI participated in this study. Heart rate (HR), blood pressure (BP) and resting pulmonary function parameters were obtained from each subject at baseline and after BWSTT. The training programme was scheduled five times per week for six weeks. The psychological status was evaluated by Beck Depression Inventory (BDI). The post-BWSTT HR value was lower than the pre-training HR value (p< 0.05). In comparison of pre- and post-BWSTT pulmonary parameters, there were only significant improvement in FVC and IC (p< 0.05). The mean BDI score was lower after BWSTT compared to before BWSTT value. As a conclusion, 6 weeks BWSTT had positive effect on heart rate and limited effect on pulmonary functions. There was an improvement in depression level of the SCI subjects. PMID- 20023353 TI - Lower limb involvement in spinal function and low back pain. AB - AIM: To evaluate evidence for involvement of the lower limb in spinal function and low back pain (LBP). DESIGN: A hypothesis based on a critical review of the relevant biomechanical and clinical literature. RESULTS: The spine resembles an inverted pendulum that supports the weight of the upper body; its stability requires a moving base that is provided by the joints of the lower limb, especially the hip. However, the sacroiliac joints are unlikely to be important for spinal function. The changing pattern of gait and development of lumbar lordosis, in early childhood, provide evidence for the inter-dependence of spinal curvature and lower limb action. Clinical signs associated with LBP may be associated with an inability to rotate the trunk about the hips. These include disorientation of the pelvis and weakness or tightness of muscles around the hip. The "sway back" posture seen in LBP involves flexion of the hip, knee and ankle to compensate for abdominal and back muscle weakness. CONCLUSIONS: In order to understand the varied clinical presentation of LBP patients, the function of the spine should be considered in the context of the whole body, especially the lower limb. PMID- 20023354 TI - Shoulder pain and functional consequences: does it differ when it is at dominant side or not? AB - PURPOSE: The shoulder complex plays an integral role in the activities of daily living and the limitation of its function can greatly affect the patient. The aim of this study was to research the functional consequences of painful conditions in the shoulder region. METHODS: Sixty five adult patients with shoulder pain for longer than one month in duration were included to the study. Dominant hands of the patients were determined. The subjects were divided into two groups: shoulder pain at the dominant upper extremity (Group I) or the nondominant one (Group II). The subjects were evaluated for shoulder pain with visual analog scale (VAS), range of motion and functional status with the Disabilities of the Arm, Shoulder, and Hand (DASH) questionnaire. RESULTS: In Group I, DASH score was significantly correlated with the VAS score measuring the pain during activity (p< 0.0001, r=0,625). In Group II, no relationship between DASH score, VAS scores and any other parameters was detected. CONCLUSION: The most important parameter disturbing the daily living of a patient with shoulder disease is the pain at the shoulder of dominant extremity occurring in active movement. PMID- 20023355 TI - A complication during caudal steroid injection. AB - BACKGROUND AND OBJECTIVE: Epidural injections in the lumbar spine are provided by caudal, lumbar interlaminar or transforaminal routes. Caudal epidural steroid injections are often used for low back pain. Fluoroscopic guidance has been frequently cited as a requirement for this procedure. In this case report, we demonstrate the importance of fluoroscopic guidance during caudal epidural injection. CASE REPORT: A 60 years old male patient was admitted to our Algology Department for low back pain. After physical examination caudal epidural steroid injection was planned. The caudal space was identified under fluoroscopic control initially using an anteroposterior projection. After the resultant epidurogram demonstrated vascular spread along the caudal epidural space the needle was withdrawn and the procedure was completed after reinserting the needle. CONCLUSION: A careful real time fluoroscopic monitoring should be applied with the injection of opaque material to minimize the risk of vascular injection. PMID- 20023356 TI - Osteopoikilosis: a case with osteopenia. AB - Osteopoikilosis is a benign osteosclerotic dysplasia of unknown origin, which is an inherited autosomal disorder. It is usually asymptomatic and may only be recognized on radiological examination. We report a 20-year-old man who suffers from back and shoulder pain during the routine physical examinations performed prior to the military recruitment. Numerous symmetric, small, ovoid and circular sclerotic bone lesions were found on the X-rays of the spine and shoulder. Dual energy X-ray absorptiometry (DXA) examination revealed osteopenia. In conclusion, typical radiographic examination of osteopoikilosis is usually diagnostic. However, it may sometimes be difficult to rule out the other diseases like osteoblastic metastatic lesions as it is an uncommon disease. Physicians should be aware of possible coexisiting osteopenia or osteoporosis as reported in our case. PMID- 20023357 TI - The effect of viscosupplementation on neuromuscular control of the knee in patients with osteoarthritis. AB - OBJECTIVE: The aim of this study was to investigate the short-term effects of intra-articular injection of hyaluronan (Hylan G-F 20) on proprioception, isokinetic muscle force, self reported pain, and functional condition in patients with knee osteoarthritis (OA). METHODS: 63 patients with stage II-III bilateral knee OA were included in this randomized, placebo controlled, and prospective study. Subjects were randomized with 42 of them into the treatment group and 21 of them into the placebo group. Hyaluronan was intraarticularly injected into both knees of the subjects which were in the treatment group, whereas physiological saline was intraarticularly injected to the subjects which were in the placebo group. Proprioception and the isokinetic muscle force measurement were performed. Visual analogue scale (VAS) and WOMAC scale were used to evaluate pain and physical function. RESULTS: Statistical analysis was performed on 120 knees of 60 patients completing the trial. The average absolute angular error (AAAE) value showing the proprioceptive error level in the treatment group was detected to be statistically significantly lower compared to placebo at the measurements performed after the 3rd injection (p = 0.02) and after one week (p = 0.01). While there was no inter-group difference in isokinetic measurements performed at 180 and 240 degrees/sec, a significant difference was detected at the measurement performed at 60 degrees/sec in favor of the treatment group (p = 0.02). Activity and resting VAS-pain values, WOMAC parameters (except the WOMAC stiffness) were detected to be significantly lower in the treatment group. Local adverse events were not reported in any patient. CONCLUSION: In this study, it was demonstrated that intraarticular injection of hyaluronan in patients with knee OA led to a short-term increase in proprioception and isokinetic muscle force, and also significant improvements in the functional conditions of patients. Long-term studies are needed. PMID- 20023358 TI - Cross cultural adaptation and reliability of the Turkish version of Amputee Body Image Scale (ABIS). AB - OBJECTIVES: The Amputee Body Image Scale (ABIS) is a self assessed questionnaire with 20 items created to measure body image perception of amputees. The questions assess how an individual perceives and feels about his or her body experience. But no Turkish version is available. The aim of this study was to cross culturally adapt the ABIS for use with Turkish speaking lower limb amputees and to determine reliability. METHODS: The sample consisted of 50 transtibial amputees. The mean age of the participants was 43.14, SD: 14.66. The ABIS has been filled by the patients for two times. RESULTS: Internal consistency of the ABIS was very high (Cronbach alpha 0.834 for test and 0.842 for retest). The test retest reliability was excellent for the ABIS (ICC = 0.939, 95% CI 0.895-0.965). CONCLUSIONS: The Turkish version of the ABIS is a reliable instrument to assess body image in lower limb amputees. PMID- 20023359 TI - Gabapentin monotherapy in patients with chronic radiculopathy: the efficacy and impact on life quality. AB - OBJECTIVE: The purpose of this, open-label, non-comparative study, was to evaluate the efficacy on quality of life and the efficacy of gabapentin monotherapy in patients with chronic radiculopathy. METHODS: Thirty-five patients with radicular pain and diagnosed as L4, L5 or S1 radiculopathy were treated with oral gabapentin from a total of 300 mg per day once up to a total of 1800 mg per day divided in 3 doses for eight-week trial period. Quality of life, functional disability and psychological mood of the patients were assessed using the Nottingham Healthy Profile (NHP), Oswestry Low Back Pain Disability Questionnaire (ODQ) and Beck Depression Inventory (BDI). RESULTS: Of the patients (n = 35), 25 were females and 10 were males (mean age: 41.8 +/- 10.4, range: 24-60 years); mean radiculopathy duration was 16.4 +/- 14.2 months (range: 3-48 months). The pain intensity at rest, quality of life, functional disability and depression scores were determined significantly improved after treatment and 4 months compared to baseline scores (p < 0.001). 1.5 points compared to baseline for at pain rest and 15 points improvement on the ODQ were obtained. CONCLUSION: Gabapentin may provide benefits in terms of alleviation of pain and overall quality of life in patients with chronic radiculopathy. PMID- 20023360 TI - Association between disc degeneration and degenerative spondylolisthesis? Pilot study. AB - OBJECTIVES: The aim of this pilot study was to test the generally believed hypothesis that intervertebral disc degeneration is a prerequisite for degenerative spondylolisthesis (DS). METHODS: This cross-sectional study was an ancillary project to the Framingham Study. A sample of 3529 participants aged 40 80 years had a CT scan performed to assess aortic calcification. 188 individuals were randomly enrolled in this study. The prevalence of intervertebral disc narrowing, facet joint osteoarthritis (FJOA) and DS were evaluated. We used the multiple logistic regressions to evaluate the association between DS as a dependent variable and FJOA, disc narrowing, age, sex and BMI as independent variables. RESULTS: There were 23 individuals (24 spinal segments) affected by DS (15 female, 8 male), mean age 62.0 +/- 6.8 years. In segments with DS, FJOA was observed and rated "severe" at 20 (83.3%) segments, "moderate" at 3 (12.5%) levels, and "mild" at 1 (4.2%) level. Intervertebral disc height was normal in 6 (25.0%), slightly decreased in 6 (25.0%), moderately decreased in 5 (20.8%) and severely decreased in 7 (29.2%) of the segments. Three (12.5%) segments with DS had severe FJOA but no apparent disc degeneration. In 9 (37.5%) segments with DS we found no or mild disc degeneration and severe FJOA. In a multiple regression analysis age, sex and FJOA, but not disc narrowing, showed significant associations with DS. CONCLUSIONS: The results of our study did not support the theory that disc degeneration necessarily precedes vertebral subluxation in DS. PMID- 20023361 TI - Determination of spondylolisthesis in low back pain by clinical evaluation. AB - AIM: Current guides recommend to evaluate the patients with low back pain complaints with initial clinical assessment and history, and to utilize radiological or other imaging technics, in case of possible diagnosis. The aim of this study was to compare the findings of radiological and clinical assessment, and validate the reliability of spondylolisthesis diagnosed with clinical assessment. This study is conducted on 100 patients with, and 30 patients without (control group) radiological diagnosis of spondylolisthesis, who had applied to Department of Physical Therapy and Rehabilitation, Haydarpasa Numune Hospital with low back pain complaints in one and a half year. Clinic assessment was consisted of 20 parameters including examinations of motor system such as, sign of slipping observed on palpation and inspection, extension of trunk and increase in lumbar lordosis. Antero-posterior, lateral, oblique and lateral flexion/extension radiographies were used for radiological assessment. Slipping degree and lumbar lordosis angle were measured. RESULTS: Women/men patients ratio was 91/9 in spondylolisthesis group and 22/8 in control group. Age of 69% of patients were 50 and over. In both groups, sciatalgia was observed in more than half of the patients, and no significant difference was detected in localization (p > 0.05). In clinical assessment, weak and drooping abdominal wall, paravertebral muscle hypertrophy, increase in lumbar lordosis, sign of slipping observed on palpation and inspection, hamstring muscle spasm, pain during lateral trunk flexion-extension tasks and during double leg raising task were found to be positively correlated with radiological assesment (p < 0.05). CONCLUSION: In our study, a systematic clinical assessment was proved to be useful in determination of possible spondylolisthesis cases. Radiological assessments are required in order to make the diagnosis clear and to determine the grade and prognosis of spondylolisthesis. Advanced imaging techniques like MRI and CT have to be used when neurological symptoms are present, and when surgical intervention is indicated. PMID- 20023362 TI - Impairment of static upright posture in subjects with undifferentiated arthritis in sacroiliac joint in conjunction with elevation of streptococcal serology. AB - BACKGROUND AND OBJECTIVES: Our latest work has demonstrated a strong correlation between the anti-streptolysin O (ASO) titer and the sacroiliac (SI) joint scintigraphy in subjects with undifferentiated arthritis [Journal of Rheumatology 34 (2007), 1746-1752]. Of a significant percentage in those subjects with sacroiliac disorder reported suffering from postural abnormality. The purpose of this study was to determine whether there was an abnormality of upright postural sway in those subjects. METHODS: All subjects who have been examined for ASO titer levels and SI joint scintigraphy were divided into two groups according to the reference level of ASO titer in our central laboratory, and were subjected to ten sway tests to assess static postural sway when they were standing upright. The comparisons of the sway parameters were analyzed by using two sample t-test for continuous variables and repeated-measures analysis of variance (ANOVA) for the degree effect and interaction effect (sloped degree x group) in varying stressful conditions (eyes open vs closed, plantar flexion or dorsiflexion of feet). RESULTS: In a total of 84 subjects, mean age was 23 years (range 18.0 36.4). Compared with the low ASO (ASO titer 116 IU/mL) group had 2.76-, 4.46- and 4.59-fold in sway area, 1.32-, 1.50- and 1.61-fold in sway velocity, and 2.02-, 1.97- and 1.70-fold in sway intensity, over the study period at 0 degrees, 10 degrees, and 20 degrees in conditions of eyes open and plantar flexion. The values of sway velocity/intensity obtained with eyes open and plantarflexion/dorsiflexion had lower intensity values when compared with those obtained in closed eyes and plantar flexion/dorsiflexion in high ASO group, but not the same as in low ASO group. Repeated-measures ANOVA showed that the sloped degree only affected the sway area in condition of eyes closed and dorsiflexion (P = 0.016), and affected the velocity/intensity in all conditions tested (all P < 0.0001). In consideration of interaction effect, the sloped degree showed significant difference in sway area in conditions of eyes open and plantar flexion/dorsiflexion (P = 0.03 and P = 0.0113), in sway velocity in most conditions tested (P < 0.05), and in sway intensity in condition of eyes open and dorsiflexion only (P = 0.0004). CONCLUSION: Subjects with high level of streptococcal serology demonstrated increased sway on all postural control measures as compared to those with low serology. Proprioceptive deficits in the SI joint might contribute to the postural impairment measured in this study. PMID- 20023363 TI - Change of quality of life due to exercise training in knee osteoarthritis: SF-36 and WOMAC. AB - The aim of this study is to assess the effect of a 12 week multicomponent exercise training program on the quality of life in females with knee osteoarthritis. Thirty four subjects, aged 50-69 years, completed a 12 week multicomponent exercise training program. The SF-36 and WOMAC were applied at baseline, at mid-training, and post-training, and the responses of the exercise group (EG) and the control group (CG) were compared. The multicomponent training program resulted in significant group differences in all domains of SF-36 (p < 0.004), while there were no group difference in WOMAC domains (p > 0.004). In the EG between mid-training and baseline period vitality score increased; physical performance and general health scores increased between mid-training and post training period; physical function, body pain, mental health, vitality, and general health scores increased between baseline and post-training period (p < 0.004). Between mid-training and baseline period of the WOMAC domains changed in the EG (p > 0.004), while all domains increased between mid-training and post training and also between baseline and post-training (p < 0.004). Changes in quality of life after multicomponent training are affected by the different evaluation techniques. PMID- 20023364 TI - The rehabilitation programme of an adult phenylketonuria with upper motor neuron involvement. AB - Phenylketonuria (PKU) is a disorder characterized by several biochemical mechanisms which may impair the brain functions in PKU, leading to neurological problems. Our case report concerns a 19 year-old man with phenylketonuria who was evaluated with the onset of stiffness following the abandonment of the phenylalanine-restricted diet. He was assessed with grade-4 spasticity according to Modified Aschworth scale. The deep tendon reflexes had increased and the plantar reflexes were positive. Knee extensions were limited due to the shortening of the hamstring muscles. Serum phenylalanine concentration was elevated and plasma vitamin B12 level was low. Cranial magnetic resonance imaging scan revealed demyelinization area in periventricular deep white matter. We administered a phenylalanine-restricted diet and a rehabilitation program. Following the treatment, spasticity was reduced to grade-1 and patient could walk without aid. This case shows that, the combination of diet, medication and a rehabilitation program is an effective treatment model on adult PKU with upper motor neuron involvement. PMID- 20023365 TI - A rare cause of a piriformis syndrome. AB - BACKGROUND AND OBJECTIVE: A less common but important cause of buttock and leg pain known as "Piriformis Syndrome". Piriformis syndrome is all intrinsic pathology of the piriformis itself, such as myofascial pain, anatomical variations, hypertrophy, and myositis ossificans or it is caused by trauma to the pelvis or buttock. In this case report we are going to present a rare cause of piriformis syndrome. CASE REPORT: Our first case was a 32 year old woman. She was referred to our pain clinic for leg pain that radiates from buttock to backside of the knee for 5 years. She did not have any problems in her history or laboratory findings. But in her lower extremity ortho roentgenogram, her leg was 2 cm short at the effected side. Second case was a 23 year old woman who had pain radiating from gluteal region to backside of the knee. In her history she had a car accident two years ago. In this accident, she had a fracture of collum femoris, and her leg was 1 cm short at the effected side. CONCLUSION: We conclude that "short leg" is one of the rare causes of piriformis syndrome and can be seen alone or with the other causes. The injection on piriformis muscle could be more effective for the patients who have PS after the "short leg" treated. PMID- 20023366 TI - A color discriminating broad range cell staining technology for early detection of cell transformation. AB - BACKGROUND: Advanced diagnostic tools stand today at the heart of successful cancer treatment. CellDetect(R) is a new histochemical staining technology that enables color discrimination between normal cells and a wide variety of neoplastic tissues. Using this technology, normal cells are colored blue/green, while neoplastic cells color red. This tinctorial difference coincides with clear morphological visualization properties, mainly in tissue samples. Here we show that the CellDetect(R) technology can be deployed to distinguish normal cells from transformed cells and most significantly detect cells in their early pre cancerous transformed state. MATERIALS AND METHODS: In tissue culture, we studied the ability of the CellDetect(R) technology to color discriminate foci in a number of two stage transformation systems as well as in a well defined cellular model for cervical cancer development, using HPV16 transformed keratinocytes. RESULTS: In all these cellular systems, the CellDetect(R) technology was able to sensitively show that all transformed cells, including pre-cancerous HPV 16 transformed cells, are colored red, whereas normal cells are colored blue/green. The staining technology was able to pick up: (i) early transformation events in the form of small type 1 foci (non-invasive, not piled up small, with parallel alignment of cells), and (ii) early HPV16 transformed cells, even prior to their ability to form colonies in soft agar. The study shows the utility of the CellDetect(R) technology in early detection of transformation events. PMID- 20023367 TI - Endothelial dysfunction aassociated with hypercortisolism is reversible in Cushing's syndrome. AB - Cushing's syndrome (CS) as represented by chronic glucocorticoid excess is associated with increased rate of cardiovascular morbidity and mortality. Because endothelial dysfunction is an early event of atherosclerosis, we investigated whether endothelial dysfunction is associated with CS and reversible. Twenty-one CS patients due to different causes were studied for vascular endothelial function by ultrasound measurement of flow-mediated vasodilation (FMD), among whom 12 patients were re-evaluated after surgical and medical treatment; 12 age- and gender-matched subjects served as control. Percent (%) FMD in CS patients (5.8+/-1.9 %) was significantly (p =0.0014) lower than that in control subjects (8.1+/-1.7 %). In CS patients, %FMD showed significant (p < 0.01) negative correlations with morning serum cortisol levels (r =-0.58) and 24-h urinary free cortisol excretion (r =-0.58). After surgical and medical treatment in CS patients, morning cortisol levels significantly (p =0.0025) decreased from 23.4 [15.6-37.3] to 10.2 [7.7-12.9] microg/dL, whereas %FMD significantly (p =0.0024) increased from 5.2+/-1.9 to 7.8+/-2.3 %; changes of %FMD after treatment significantly (p =0.0004) and inversely correlated with those of morning cortisol levels (r =-0.85), but not with those of body mass index, blood pressure, glycemic or lipid profiles. Taken together, the present study clearly revealed that endothelial dysfunction in CS patients is related to hypercortisolemia and reversible after treatment, suggesting the possible role of cortisol excess in the development of endothelial dysfunction, thereby possibly leading to increased cardiovascular complications. PMID- 20023368 TI - The epidemiology of appendicitis and appendectomy in South Korea: national registry data. AB - BACKGROUND: Appendicitis is one of the most frequent acute surgical conditions of the abdomen, and appendectomy is one of the most commonly performed operations in the world. However, epidemiological data on appendicitis have not been reported for South Korean or East Asian populations. METHODS: We analyzed the epidemiological features and lifetime risk of appendicitis and appendectomy in South Korea using data collected for the national health insurance database from 2005 through 2007. RESULTS: Appendectomy was performed in 59.70% of inpatients diagnosed with appendicitis. The overall incidences of appendicitis, total appendectomy, and perforated appendectomy were 22.71, 13.56, and 2.91 per 10 000 population per year, respectively. The incidence of appendicitis and appendectomy showed clear seasonality, with a peak in summer. The standardized lifetime risks of appendicitis and appendectomy were constant from 2005 through 2007. A life table model suggests that the lifetime risk of appendicitis is 16.33% for males and 16.34% for females, and that the lifetime risk of appendectomy is 9.89% for males and 9.61% for females. CONCLUSIONS: As compared to results obtained in research on Western populations, appendicitis and appendectomy had a similar perforation rate and seasonality, but a higher overall incidence, in South Koreans. Between 2005 and 2007, the incidence of appendicitis and appendectomy was constant. Overall, an estimated 15 incidental appendectomies are performed to prevent 1 inpatient with suspected appendicitis, and 26 incidental appendectomies are performed to prevent 1 appendectomy. Incidental appendectomy may have greater preventive value in Koreans. PMID- 20023369 TI - [Mechanism for the induction of host immune response by virulence factors of Listeria monocytogenes]. PMID- 20023371 TI - Treatment for glioma recurrence: new hope? PMID- 20023374 TI - An EGOcentric view of TORC1 signaling. PMID- 20023372 TI - Stimulation or Inhibition: Conflicting evidence for (+/-)-catechin's role as a chemical facilitator and disease protecting agent. AB - The occurrence of plant hormesis is a poorly understood phenomenon, wherein low doses of phytotoxins unusually promote growth responses in higher plants. In contrast, negative plant-plant interactions mediated through secreted small molecular weight compounds initiate growth inhibitory responses. Studies related to (+/-)-catechin mediated allelopathy have transpired both novel information and generated significant controversy. Specifically, studies related to the phytotoxicity responses mediated by (+/-)-catechins have been seriously debated. The pronged opinion that (+/-)-catechin is phytotoxic versus non-phytotoxic relies more on the target plant systems and the conditions used to test phytotoxic responses. It is reported that lower than MIC dosage supplementation of (+/-)-catechin could promote growth responses in the model plant Arabidopsis thaliana. Furthermore, it was shown that sub-MIC levels of (+/-)-catechin supplementation leads to elicitation of disease resistance against Pseudomonas syringae DC3000 (hereafter DC3000). Intrigued by the unique hormesis response observed, we tested whether (+/-)-catechin indeed promotes growth responses in A. thaliana. In our hands, we observed no growth promotion responses of (+/-) catechin against A. thaliana under in vitro or in soil conditions. We also evaluated the previously reported disease protecting properties of (+/-)-catechin in A. thaliana against DC3000. The systematic observations to evaluate disease protecting properties entailing colony counts, disease incidences and loss of chlorophyll studies showed no disease protecting properties of (+/-)-catechin. The transcriptional response for a marker pathogenesis related PR1 defense gene showed no induction post (+/-)-catechin supplementation. The cell death genes (ACD2 and CAD1) associated with programmed cell death revealed unchanged expression levels in plants treated with sub-MIC levels of (+/-)-catechin. Further, we report supplementation of sub-MIC levels of (+/-)-catechin negates any change in the expression of an auxin responsive gene. Our results refute the previous claims of growth and defense inducing effects of (+/-)-catechin, thus suggesting that a thorough reexamination is required to evaluate the hormetic effect of (+/-)-catechin under both controlled and natural conditions. PMID- 20023376 TI - An emerging role for class I bHLH E2-2 proteins in EMT regulation and tumor progression. AB - EMT is a complex process whereby cells lose cell-cell interactions and other epithelial properties whilst acquiring a migratory and mesenchymal phenotype. EMT is presently recognized as an important even for tumor invasion and metastasis. Functional E-cadherin loss is a hallmark of EMT and required for tumor invasion in the majority of carcinomas. Transcriptional downregulation is one of the major mechanisms for E-cadherin suppression in carcinomas. In the last decade several E cadherin repressors, belonging to different transcriptional families, have been identified that, importantly, also act as potent EMT inducers. One of the last additions to EMT regulators are the class I bHLH factors E2-2 (also known as TCF4). However, the hierarchical and functional interrelations between the different EMT inducers are still poorly understood. Here, we comment on the new and so far unrecognized function of E2-2 factors in EMT and discuss on the potential interactions among various EMT inducers. Emerging evidence supporting the participation of TCF4 in human malignancies is also discussed. Thus, increasing understanding of EMT and its regulators is providing meaningful insights into the present knowledge on tumor progression. PMID- 20023377 TI - Foxo1 in hepatic lipid metabolism. PMID- 20023380 TI - A cautionary tale on genetic screens based on a gain-of-expression approach: The case of LanB1. AB - Gain of function screens have being frequently used to search for genes affecting a particular adult character or developmental process. These experiments are made possible by the adoption of the Gal4/UAS system to flies, and by the design of P elements bearing UAS sequences. We recently published two screens in which a large number of newly generated P-UAS insertions were crossed with Gal4 drivers expressed in the pupal veins and in the central region of the wing disc. From the data obtained in these and other screens, it seems that a gain-of-function phenotype is a rare occurrence observed only for about 5-8% of insertion sites. Insertions affecting the expression of signaling molecules were particularly enriched in the screens. In contrast, gain-of-function phenotypes due to insertions not belonging to this class appear to be caused by multiple protein specific mechanisms that could only be unraveled after extensive analysis. We present some data concerning the overexpression of LamB1, a gene encoding the B subunit of Laminin trimers in Drosophila, and show that Notch protein subcellular localization and signaling is compromised in cells overexpressing LanB1. PMID- 20023378 TI - Blocking ovarian cancer progression by targeting tumor microenvironmental leukocytes. AB - Current therapies for metastatic ovarian carcinoma are based on surgical debulking followed by chemotherapy. After more than three decades implementing treatments that selectively target the tumor cell, the 5-year survival rate for metastatic ovarian cancer patients is still lower than 30%. Novel strategies are therefore urgently needed to complement classical treatments for this malignancy. Recently, leukocytes in the ovarian cancer microenvironment such as regulatory T cells and immature pro-angiogenic/tolerogenic myeloid cells have been demonstrated to play a fundamental role in tumor progression. This review focuses on our recent understanding of the potential of eliminating and/or modulating the phenotype of these leukocytes in vivo and in situ as a novel intervention to complement standard ovarian cancer treatments. The significant effects of targeting these crucial microenvironmental players on cancer vascularization, local tumor growth, distal metastatic spreading and spontaneous anti-tumor immune responses are discussed. PMID- 20023379 TI - A systematic approach to understand the functional consequences of non-protein coding risk regions. AB - A primary goal of genetic association studies is to elucidate genes and novel biological mechanisms involved in disease. Recently, genome-wide association studies have identified many common genetic variants that are significantly associated with complex diseases such as cancer. In contrast to Mendelian disorders, a sizable fraction of the variants lies outside known protein-coding regions; therefore, understanding their biological consequences presents a major challenge in human genetics. Here we describe an integrated framework to allow non-protein coding loci to be annotated with respect to regulatory functions. This will facilitate identification of target genes as well as prioritize variants for functional testing. PMID- 20023381 TI - Identification of a functional nuclear export sequence in diacylglycerol kinase zeta. AB - Diacylglycerol kinases (DGKs) are key regulators of diacylglycerol-dependent signaling pathways. Among the 10 DGK isoforms, DGK-zeta is the only nuclear form that contains a nuclear localization signal. Here, by site-directed mutagenesis, we showed that DGK-zeta also displays a functional independent nuclear export signal (NES) sequence between the amino acid residues 362-370. Indeed, the NES mutant forms of DGK-zeta accumulated in the nucleus to a much greater extent than wildtype DGK-zeta. Moreover, treatment with leptomycin B, an inhibitor of leucine rich type NES, resulted in accumulation of both endogenous and ectopically expressed DGK-zeta in the nucleus, demonstrating that nuclear export of DGK-zeta is chromosome regional maintenance protein 1 (CRM1)-dependent. Previously, we reported that nuclear DGK-zeta is a negative regulator of cell cycle progression in C2C12 mouse myoblasts. In this paper, we documented that enhancement of DGK zeta nuclear localization by NES sequence mutation, increases G(0)/G(1) block in C2C12 cells. Overall, our data demonstrate that DGK-zeta export from nucleus to cytoplasm is regulated by a leucine-rich NES through the exportin CRM1 and suggest that the nuclear localization of DGK-zeta could finely tune its function as a regulator of G(1)/S cell cycle transition. PMID- 20023382 TI - Expression of CD133 correlates with differentiation of human colon cancer cells. AB - CD133 has been identified as a cancer stem cell marker in colon and several other cancers, but its function is still unknown. We examined the CD133 expression in 44 human cancer cell lines, and found five of the 8 positive lines were from colon cancer. The CD133 positive subpopulation of colon cancer cells showed more vigorous growth and lower differentiation. Induction of differentiation reduced the CD133-positive population. Knockdown of CD133 expression in colon cancer cells could not induce cellular differentiation. Care must be taken if CD133 is used as the only marker of cancer stem cells in colon cancer, especially in established cell lines. CD133 negatively correlates with cell differentiation, but it is not a regulator of differentiation. PMID- 20023383 TI - Crosstalk between planar cell polarity signaling and miR-8 control of NHERF1 mediated actin reorganization. AB - The response to osmotic stress in developing zebrafish embryos requires proper apical patterning and trafficking of transmembrane ion transporters in ionocytes, specialized cells of the epidermis. The miR-8 family of miRNAs plays a key role in this process by precisely regulating the activity of NHERF1, a regulator of sodium hydrogen exchange that also serves as an adaptor protein linked to the actin cytoskeleton. We have discovered that NHERF1 activity is also coupled to Planar Cell Polarity (PCP) signaling in the zebrafish epidermis. Loss of NHERF1 in wild type fish disrupts actin organization but the observed defects can be largely restored when combined with mutants in the PCP pathway. We propose that proper apical patterning depends on input and coordination between PCP signaling and the response to stress. PMID- 20023384 TI - Defective autophagy control by the p53 rheostat in cancer. AB - Autophagy is a finely regulated, lysosomal catabolic pathway that contributes to the turnover of long-lived proteins and to the elimination of old/damaged organelles. Autophagy exerts bona fide oncosuppressive functions by: (1) limiting chromosomal instability; (2) reducing potentially mutagenic oxidative stress; and (3) restraining intratumoral necrosis and local inflammation. Defective autophagy constitutes a hallmark of cancer cells together with: (1) provision of autonomous growth signals;, (2) insensitivity to antiproliferative stimuli; (3) disabled apoptosis; (4) limitless replication; (5) production of angiogenic factors; (6) tissue invasion with metastasis; (7) avoidance of the immune response; and (8) enhanced anabolism. p53 is the best-known human oncosuppressor protein, and its genetic/epigenetic inactivation has been observed in more than 50% of all human cancers. p53 mostly mediates tumor suppression by transactivating pro-apoptotic and cell cycle arresting genes, but also by favoring mitochondrial apoptosis in a transcription-independent fashion, by modulating metabolic circuitries and by regulating autophagy. p53 mutations (or epigenetic changes) that simultaneously abolish its pro-apoptotic and autophagy-inhibitory functions behave as "multi hit" events, as opposed to "single-hit" mutations that only affect the classical (pro-apoptotic and/or cell cycle-arresting) functions of the p53 system. We speculate that, in this latter case, additional genetic/epigenetic events resulting in disabled autophagy are likely to contribute to accelerated oncogenesis. PMID- 20023385 TI - Sterol biosynthesis in oomycete pathogens. AB - Oomycetes are a diverse group of filamentous eukaryotic microbes comprising devastating animal and plant pathogens. They share many characteristics with fungi, including polarized hyphal extension and production of spores, but phylogenetics studies have clearly placed oomycetes outside the fungal kingdom, in the kingdom Stramenopila which also includes marine organisms such as diatoms and brown algae. Oomycetes display various specific biochemical features, including sterol metabolism. Sterols are essential isoprenoid compounds involved in membrane function and hormone signaling. Oomycetes belonging to Peronosporales, such as Phytophthora sp., are unable to synthesize their own sterols and must acquire them from their plant or animal hosts. In contrast, a combination of biochemical and molecular approaches allowed us to decipher a nearly complete sterol biosynthetic pathway leading to fucosterol in the legume pathogen Aphanomyces euteiches, an oomycete belonging to Saprolegniales. Importantly, sterol demethylase, a key enzyme from this pathway, is susceptible to chemicals widely used in agriculture and medicine as antifungal drugs, suggesting that similar products could be used against plant and animal diseases caused by Saprolegniales. PMID- 20023387 TI - Modulating c-Abl nuclear activity as a strategy to preserve female fertility. PMID- 20023388 TI - Pinpointing Pin1 in non-small cell lung carcinoma. AB - Factors that promote tumorigenesis encompass a wide array of proteins that include transcription factors, signal transduction molecules, negative regulators of tumor suppressors, transmembrane receptors and enzymes involved in a variety of processes. Identifying these factors remains one of the most pertinent issues in cancer research, as they can be used as both diagnostic markers and therapeutic targets. The ability to characterize tumors based on molecular markers will enable one to easily assess tumor severity, select a method of treatment and ultimately improve patient outcome. While the treatment of primary tumors has improved by leaps and bounds over the past decade, metastases continue to plague both patients and the research community. Along these lines, markers that predict metastatic potential will help to both limit secondary tumor growths and improve patient outcome. PMID- 20023389 TI - Linking rDNA transcription to the cellular energy supply. PMID- 20023390 TI - Artophagy: the art of autophagy--the Cvt pathway. AB - Science informs art, and art informs science. Both processes involve creativity and imagination, and collaboration between scientists and artists often leads to new insights in both fields. We took advantage of the power of artistic imagery to demonstrate a dynamic cellular process, autophagy. In particular, we depicted the cytoplasm to vacuole targeting pathway, which involves dynamic membrane rearrangements to sequester a specific cargo via an autophagy-related process. By depicting this event in the context of a crowded cellular milieu, we hoped to stimulate researchers to consider aspects of the process that might be overlooked in the overly simplistic schematic drawing that typify most scientific models. PMID- 20023391 TI - Plant SUN domain proteins: components of putative plant LINC complexes? PMID- 20023392 TI - AMPK: A metabolic checkpoint that regulates the growth of EGFR activated glioblastomas. PMID- 20023393 TI - The PP2C-SnRK2 complex: the central regulator of an abscisic acid signaling pathway. PMID- 20023394 TI - Phospho-DeltaNp63alpha/NF-Y protein complex transcriptionally regulates DDIT3 expression in squamous cell carcinoma cells upon cisplatin exposure. AB - Cisplatin remains the most important chemotherapeutic agent for patients with human head and neck cancer. However, tumor cells often develop resistance to cisplatin-induced apoptosis. We previously found that head and neck squamous cell carcinoma (HNSCC) cells exposed to cisplatin display a marked ATM-induced phosphorylation of DeltaNp63alpha. However, the mutated Np63-S385G failed to undergo phosphorylation by ATM kinase. We used HNSCC cell lines expressing the wild type DeltaNp63alpha or mutated DeltaNp63alpha-S385G to determine the effect of S385G mutation on the DeltaNp63alpha transcriptional activity and protein protein interactions. The S385G mutation in DeltaNp63alpha dramatically abolished the upregulation/downregulation of downstream gene targets and the binding of DeltaNp63alpha-S385G to certain promoters. In contrast to the non-phosphorylated DeltaNp63alpha-S385G, the phospho-DeltaNp63alpha forms protein-protein complexes with NF-YA transcription factor and regulates the transcription of DDIT3 gene implicated in the programmed cell death of HNSCC cells upon cisplatin exposure. We suggest that the transcriptional activation of DeltaNp63alpha through its phosphorylation by ATM kinase in HNSCC cells exposed to cisplatin is a critical step in the subsequent sensitivity of certain human head and neck cancers to platinum therapy. PMID- 20023395 TI - Co-localization of mitochondria with chloroplasts is a light-dependent reversible response. PMID- 20023397 TI - An adult tissue-specific stem cell molecular phenotype is activated in epithelial cancer stem cells and correlated to patient outcome. AB - Recent studies have shown that embryonic stem cell-like molecular phenotypes are commonly activated in human epithelial primary tumors and are linked to adverse patient prognosis.(1,2) However it remains unclear whether these correlations to outcome are linked to the differentiation status of the human primary tumors(1) or represent molecular reminiscences of epithelial cancer stem cells.(2) In addition, while it has been demonstrated that leukemic cancer stem cells re acquire an embryonic stem cell-like phenotype,(3,4) the molecular basis of stem cell function in epithelial cancer stem cells has not been investigated. Here we show that a normal adult tissue-specific stem cell molecular phenotype is commonly activated in epithelial cancer stem cells and for the first time provide evidence that enrichment in cancer stem cells-specific molecular signatures are correlated to highly aggressive tumor phenotypes in human epithelial cancers. PMID- 20023398 TI - Hes1 oscillation: making variable choices for stem cell differentiation. PMID- 20023399 TI - Dual regulation of iron deficiency response mediated by the transcription factor IDEF1. PMID- 20023400 TI - Mechanistic insights into mRNA export through structures of Dbp5. AB - Nuclear export of mRNA is a critical event in mRNA biogenesis. Passage of mature messenger ribonucleoproteins (mRNPs) through nuclear pore complexes (NPCs) is facilitated by the Mex67/Mtr2 heterodimer. At the NPC cytoplasmic face, the DEAD box RNA helicase Dbp5 remodels mRNPs by removing Mex67/Mtr2. This remodeling process prevents mRNPs from returning to the nucleus, thereby imposing unidirectionality on mRNA export. Biochemical studies show that Gle1 and inositol hexaphosphate (IP6) activate Dbp5's ATPase activity at the cytoplasmic face of NPC, therefore providing critical spatial regulation of mRNP remodeling during directional transport. Recent structural studies on Dbp5 in free form and in complex with its ligands (ADP, AMPPNP/RNA) as well as with cytoplasmic nucleoporin Nup214 reveal that the binding of ADP or AMPPNP/RNA induces large conformational changes of Dbp5, and RNA and NUP214 bind to Dbp5 in a mutually exclusive manner. These structural data combined with complementary functional analysis significantly advance our understanding on the mechanism governing mRNA export albeit some key issues remain elusive. PMID- 20023402 TI - Identification of an RNA-dependent RNA polymerase in Drosophila establishes a common theme in RNA silencing. AB - In lower eukaryotes, such as A. thaliana, C. elegans, S. pombe and N. crassa, RNA dependent RNA polymerase (RdRP) is a required component of the RNA silencing pathway. Remarkably, even though robust RNA silencing occurs in Drosophila in response to exogenous dsRNA and siRNAs, no RdRP homolog has been identified in the Drosophila genome or in any other higher eukaryote characteristic of the known cellular RdRPs. We showed recently that the largest subunit of the Drosophila RNA polymerase II core elongator complex, called D-elp1, has RdRP activity capable of using unprimed or primed synthesis mechanisms to convert single stranded RNA templates into double stranded RNA (dsRNA) that can be cleaved by Dcr-2. Loss of D-elp1 inhibits both siRNA and dsRNA directed RNAi in S2 cells but does not affect micro RNA targeting. Transposon RNA levels also increase with the loss of D-elp1 while the corresponding endo siRNAs, critical for transposon suppression, are dramatically reduced and this is correlated with a reduction in transposon antisense RNA levels. D-elp1 associates tightly with Dicer-2, similar to the Dicer-RdRP interaction observed in lower eukaryotes. With the exception of S. cerevisiae, which lacks the RNAi machinery altogether, RdRP activity is conserved in the elp1 homologs from S. pombe to human. This commentary focuses on the importance and universality of RdRP in RNA silencing. PMID- 20023401 TI - Thermo and pH stable ATP-independent chaperone activity of heat-inducible Hsp70 from Pennisetum glaucum. AB - Heat shock proteins (Hsps) are a class of molecular chaperones that play an essential role in preserving cellular functions under stressful conditions. The over production of recombinant proteins often causes cellular stress that results in aggregation/misfolding of proteins, which sometimes leads to the formation of inclusion bodies. Here we report the cloning and characterization of heat inducible PgHsp70 from Pennisetum glaucum, a heat and drought tolerant plant that showed stability and chaperone activity at elevated temperatures. The predicted amino acid sequence of PgHsp70 revealed a high homology with Hsp70 from other plants, and the overall 3D structure homology modeling is similar to that of the constitutively expressed bovine cytosolic Heat Shock Cognate (HSC)-70. The purified recombinant protein had an apparent molecular mass of 70 kDa and displayed optimal chaperone activity at 50 degrees C, and pH 8.0. Under these conditions, the T(1/2) of PgHsp70 increased from 10 to 15 h in the presence of glycerol. The PgHsp70 exhibited a higher chaperone activity towards glutamate dehydrogenase than alcohol dehydrogenase. The expression of recombinant carbonic anhydrase (CA) in E. coli in a catalytically active soluble form rather than in inclusion bodies was made feasible by co-expression of PgHsp70. Circular dichroism (CD) studies of the recombinant PgHsp70 did not reveal any discernible changes in the alpha-helix content, with increase in temperature from 35 to 85 degrees C, thus suggesting a critical role of alpha-helix content in maintaining the chaperone activity. PMID- 20023403 TI - The plant as a biomechatronic system. AB - Our vision of plants is changing dramatically: from insensitive and static objects to complex living beings able to sense the environment and to use the information collected to adapt their behaviour. At all times humans imitate ideas and concepts from nature to resolve technological problems. Solutions coming from plants have the potential to face challenges and difficulties of modern engineering design. Characteristic concepts of the plant world such as reiteration, modularity and swarm behaviour could be of great help resolving technological problems. On the other hand a biorobotic approach would facilitate the resolution of many biological problems. In this paper, the concept of a plant inspired robot is proposed for the investigation of both biological and technological issues. PMID- 20023404 TI - Targeting the checkpoint kinase Chk1 in cancer therapy. AB - A paramount objective of the eukaryotic cell division cycle is to overcome numerous internal and external insults to faithfully duplicate the genetic information once per every cycle. This is carried out by elaborate networks of genome surveillance signaling pathways, termed replication checkpoints. Central to replication checkpoints are two protein kinases, the upstream kinase ATR, and its downstream target kinase, Chk1. When the DNA replication process is interrupted, the ATR-Chk1 pathway transmits signals to delay cell cycle progression, and to maintain fork viability so that DNA duplication can resume after the initial damage is corrected. Previous studies showed that replicative stress not only activated Chk1, but also triggered the ubiquitin-dependent destruction of Chk1 in cultured human cells. In a recent study, we identified the F-box protein, Fbx6, as the mediator that regulates Chk1 ubiquitination and degradation in both normally cycling cells and during replication stress. We further showed that expression levels of Chk1 and Fbx6 exhibited an overall inverse correlation in both cultured cancer cell lines and in breast tumor tissues, and that defects in Chk1 degradation, for instance, due to reduced expression of Fbx6, rendered tumor cells resistant to anticancer treatment. Here we highlight those findings and their implications in the replication checkpoint and cellular sensitivity to cancer therapies. PMID- 20023405 TI - Mechanisms of microRNA-mediated auxin signaling inferred from the rice mutant osaxr. AB - Auxin, known as the central hormone, plays essential roles in plant growth and development. In auxin signaling pathways, the tiny RNA molecules, i.e., microRNAs (miRNAs), show their strong potential in modulating the auxin signal transduction. Recently, we isolated a novel auxin resistant rice mutant osaxr (Oryza sativa auxin resistant) that exhibited plethoric root defects. Microarray experiments were carried out to investigate the expression patterns of both the miRNAs and the protein-coding genes in osaxr. A number of miRNAs showed reduced auxin sensitivity in osaxr compared with the wild type (WT), which may contribute to the auxin-resistant phenotype of the mutant. Auxin response elements (AuxREs) were demonstrated to be more frequently present in the promoters of auxin-related miRNAs. In our previous report, a comparative analysis of miRNA and protein coding gene expression datasets uncovered a number of reciprocally expressed miRNA-target pairs. A feedback circuit between miRNA and auxin response factor (ARF) was then proposed. Here, we will discuss in-depth some points raised in the previous report, in particular, the organ-specific expression patterns of miR164, the feedback regulatory model between miR167 and certain ARFs, and the potential signal interactions between auxin and nutrition or stress that are mediated by miRNAs in rice roots. PMID- 20023406 TI - Regulation of the female mouse germ cell cycle during entry into meiosis. AB - During mouse embryonic development germ cells proliferate extensively until they commit to the male or female pathway and arrest in mitosis or meiosis respectively. Whilst the transition of female germ cells exiting the mitotic cell cycle and entering meiosis is well defined histologically, the essential cell cycle proteins involved in this process have remained unresolved. Using flow cytometry we have examined the entry of female germ cells into meiosis, their termination of DNA synthesis and entry into prophase I. Analysis of key G(2)/M cell cycle proteins revealed that entry into meiosis and cell cycle exit at G(2)/M involves repression of G(2)/M promoting Cyclin B1, coincident upregulation of G(2)/M repressing Cyclin B3 and robust establishment of the ATM/CHK2 pathway. By contrast we show that the ATR/CHK1 pathway is activated in male and female germ cells. This data indicates that an important G(2)/M surveillance mechanism operates during germ cell proliferation and that passage into meiotic G(2)/M involves the combined repression of G(2)/M through Cyclin B3 and activation of the key G(2)/M checkpoint regulatory network modulated through ATM and CHK2. This work shows that the core regulatory machinery that controls G(2)/M progression in mitotic cells is activated in female mouse germ cells as they enter meiosis. PMID- 20023407 TI - Reactive oxygen species derived from impaired quality control of photosystem II are irrelevant to plasma-membrane NADPH oxidases. AB - Protein quality control plays an important role in the photosynthetic apparatus because its components receive excess light energy and are susceptible to photooxidative damage. In chloroplasts, photodamage is targeted to the D1 protein of Photosystem II (PSII). The coordinated PSII repair cycle (PSII disassembly, D1 degradation and synthesis, and PSII reassembly) is necessary to mitigate photoinhibition. A thylakoid protease FtsH, which is formed predominantly as a heteromeric complex with two isoforms of FtsH2 and FtsH5 in Arabidopsis, is the major protease involved in PSII repair. A mutant lacking FtsH2 (termed var2) shows compromised D1 degradation. Furthermore, var2 accumulates high levels of chloroplastic reactive oxygen species (cpROS), reflecting photooxidative stress without functional PSII repair. To examine if the cpROS produced in var2 are connected to a ROS signaling pathway mediated by plasma membrane NADPH oxidase (encoded by AtRbohD or AtRbohF), we generated mutants in which either Rboh gene was inactivated under var2 background. Lack of NADPH oxidases had little or no impact on cpROS accumulation. It seems unlikely that cpROS in var2 activate plasma membrane NADPH oxidases to enhance ROS production and the signaling pathway. Mutants that are defective in PSII repair might be valuable for investigating cpROS and their physiological roles. PMID- 20023408 TI - How and where are nonsense mRNAs degraded in mammalian cells? AB - The nonsense-mediated mRNA decay (NMD) pathway is responsible for the rapid degradation of eukaryotic mRNAs on which ribosomes fail to terminate translation properly. NMD thereby contributes to the elimination of aberrant mRNAs, improving the fidelity of gene expression, but also serves to regulate gene expression at the post-transcriptional level. Here we discuss recent evidence as to how and where mRNAs targeted to NMD are degraded in human cells. We discuss accumulating evidence that the decay step of human NMD can be initiated by two different mechanisms: either by SMG6-mediated endonucleolytic cleavage near the aberrant stop codon, or by deadenylation and decapping. While there is evidence that mRNAs targeted for NMD have the capacity to accumulate with other translationally repressed mRNAs in P-bodies, there is currently no evidence that this is required for the degradation of the NMD substrate. It therefore remains an open question whether NMD in human cells is restricted to a particular cellular location or whether it can be initiated wherever translation of the NMD substrate takes place. PMID- 20023410 TI - The life span-prolonging effect of sirtuin-1 is mediated by autophagy. AB - The life span of various model organisms can be extended by caloric restriction as well as by autophagy-inducing pharmacological agents. Life span-prolonging effects have also been observed in yeast cells, nematodes and flies upon the overexpression of the deacetylase Sirtuin-1. Intrigued by these observations and by the established link between caloric restriction and Sirtuin-1 activation, we decided to investigate the putative implication of Sirtuin-1 in the response of human cancer cells and Caenorhabditis elegans to multiple triggers of autophagy. Our data indicate that the activation of Sirtuin-1 (by the pharmacological agent resveratrol and/or genetic means) per se ignites autophagy, and that Sirtuin-1 is required for the autophagic response to nutrient deprivation, in both human and nematode cells, but not for autophagy triggered by downstream signals such as the inhibition of mTOR or p53. Since the life spanextending effects of Sirtuin-1 activators are lost in autophagy-deficient C. elegans, our results suggest that caloric restriction and resveratrol extend longevity, at least in experimental settings, by activating autophagy. PMID- 20023409 TI - Apolipoprotein D: an overview of its role in aging and age-related diseases. PMID- 20023411 TI - Probing the actin-auxin oscillator. AB - The directional transport of the plant hormone auxin depends on transcellular gradients of auxin-efflux carriers that continuously cycle between plasma membrane and intracellular compartments. This cycling has been proposed to depend on actin filaments. However, the role of actin for the polarity of auxin transport has been disputed. To get insight into this question, actin bundling was induced by overexpression of the actin-binding domain of talin in tobacco BY 2 cells and in rice plants. This bundling can be reverted by addition of auxins, which allows to address the role of actin organization on the flux of auxin. In both systems, the reversion of a normal actin configuration can be restored by addition of exogenous auxins and this fully restores the respective auxin dependent functions. These findings lead to a model of a self-referring regulatory circuit between polar auxin transport and actin organization. To further dissect the actin-auxin oscillator, we used photoactivated release of caged auxin in tobacco cells to demonstrate that auxin gradients can be manipulated at a subcellular level. PMID- 20023412 TI - DNA damage signaling is activated during cancer progression in human colorectal carcinoma. AB - PURPOSE: Recent studies have shown that the DNA damage response (DDR) is activated in precancerous lesions, suggesting that neoplastic cells may avoid apoptosis by impairing the DDR which acts as a barrier against tumor progression. To define the role of the DDR pathway in human colorectal carcinoma, we investigated the level of phosphorylated proteins of the DDR pathway. RESULTS: Immunostaining for pATM, gammaH2AX and pChk2 revealed that all were significantly expressed during tumor progression in advanced carcinoma (vs. normal tissue for pATM [p < 0.05]; vs. normal and adenoma for gammaH2AX [p < 0.05]; and vs. normal tissue for pChk2 [p < 0.05]. Western blot analysis of gammaH2AX and pChk2 revealed that their level increased gradually during tumor progression and was maximal in advanced carcinoma (vs. normal tissue; p < 0.05). No apoptotic cells were found in any tissue sample. EXPERIMENTAL DESIGN: Colorectal tissue samples were obtained at the time of surgery, from 55 patients at two hospitals. The tissues were classified into four groups according to pathology: normal mucosa, adenoma, early carcinoma and advanced carcinoma. We evaluated phosphorylated ataxia telangiectasia mutated (pATM), phosphorylated H2AX (gammaH2AX) and Chk2 (pChk2) protein levels by immunohistochemistry and western blot analysis. We also evaluated apoptosis by the TUNEL assay. CONCLUSIONS: The DDR pathway was activated during cancer progression, but no apoptosis was detected, even among the cells with activated DDR. It is likely that activation of DDR was induced by stress signaling as a consequence of oxidative, replication and mechanical stresses occurring during growth and expansion of the colorectal cancer. PMID- 20023413 TI - The evolution of the regulatory mechanism of chloroplast division. PMID- 20023414 TI - Colorectal tumor derived fibronectin alternatively spliced EDA domain exserts lymphangiogenic effect on human lymphatic endothelial cells. AB - The objective of this study was to investigate whether tumor derived fibronectin alternatively spliced EDA domain has a lymphangiogenic potency on human lymphatic endothelial cells (LECs) in tumor generation to facilitate tumor lymphatic metastasis. LECs were cultured in three-dimentional culture system and treated with SW480 supernant which was highly rich in EDA, the result demonstrated that SW480 supernant could facilitate tube-like formations of LECs evidently when compared with controls. Integrinalpha9 was identified by immunofluorescence to be a specific receptor for EDA because we found co-locozation of EDA and integrinalpha9 on LECs as well as significant upregulation of integrinalpha9 in SW480 supernant treated group. Western blot and immunofluorescence revealed that EDA also had important roles accommodating the expressions of some key regulators of lymphangiogenesis such as Prox1 and F-actin so as to facilitate motility and sprouting of LECs. In addition, it had been confirmed that all of these effects could be inhibited markedly by EDA antibody (IST-9). Based on these findings, we assert that EDA derived from tumor cells has an important role in facilitating lymphangiogenesis of malignant tumor. Furthermore, EDA pathway may provide a potent target for tumor lymphatic metastasis therapy. PMID- 20023415 TI - Pivotal role of calcium/calmodulin-dependent protein kinase II in ER stress induced apoptosis. PMID- 20023416 TI - Magnesium-sensing riboswitches in bacteria. AB - All living organisms require mechanisms for coordination of intracellular metal concentrations. Many types of metal-sensing regulatory proteins have been described previously. Data in recent years have revealed that posttranscriptional mechanisms are also utilized for control of metal ion homeostasis in bacteria. In particular, two classes of RNA structural elements have been discovered to coordinate magnesium-induced structural transitions with expression levels of downstream genes. We discuss these types of regulatory RNAs herein and compare them to other classes of riboswitches that respond instead to small organic metabolites. PMID- 20023417 TI - KN-93 inhibits androgen receptor activity and induces cell death irrespective of p53 and Akt status in prostate cancer. AB - It has been suggested that the downregulation of AR expression should be considered the principal strategy for the treatment of hormone-refractory prostate cancer. We have previously shown that inhibition of AR induced PI3K independent activation of Akt that was mediated by CaMKII. In this study, we found that the CaMKII inhibitor KN-93 has a broader effect on apoptosis than just inhibition of CaMKII: first, KN-93 inhibits AR activity and induces cell death in PCa cells after androgen deprivation when many other drugs fail to kill prostate cancer cells; second, KN-93 inhibits expression of the anti-apoptotic protein Mcl 1 and induces expression of the pro-apoptotic protein PUMA; third, KN-93-mediated cell death is p53-independent; and fourth, KN-93 induces the generation of ROS. The ROS induction allows KN-93 to circumvent the activation of Akt, which occurs in prostate cancer cells under androgen deprivation, since Akt could not inhibit ROS-mediated apoptosis. KN-93 also synergistically induces cell death in combination with low doses of doxorubicin and converts the phenotype of prostate cancer cells from TRAIL-resistant to -sensitive. These data suggest that KN-93 could be used for novel therapeutic approaches when hormonal therapy has failed. PMID- 20023418 TI - FANCD2: A DNA binding protein regulated by MRE11-RAD50-NBS1. PMID- 20023419 TI - Use of modified RECIST criteria to improve response assessment in targeted therapies: challenges and opportunities. PMID- 20023420 TI - Defective autophagy associated with LC3 puncta in epothilone-resistant cancer cells. AB - Autophagy is commonly characterized by the redistribution of the microtubule associated light chain 3 (LC3) protein into cytoplasmic puncta, coinciding with its lipidation, as well as by a decrease in the abundance of autophagic substrates including p62 and ubiquitinylated proteins. Here, we describe a cell line, A549-B480, which, in contrast to its parental A549 line, exhibits massive accumulation of LC3 (or a GFP-LC3 fusion protein) in cytoplasmic puncta. These puncta co-localize with accumulated p62 and ubiquitinylated proteins, yet are not enwrapped by membranes. Indeed, LC3 is not lipidated in A549-B480, even when these cells are cultured in conditions in which A549 cells would develop autophagy. A549-B480 cells have been selected for their resistance against the microtubule-stabilizing agent epothilone B and actually require the continuous presence of epothilone B for their survival. Parental A549 cells treated with epothilone B manifested all signs of bona fide autophagy. In contrast, the autophagic program of A549-B480 was defective, irrespective of the absence or presence of epothilone B, and correlated with the complete absence of Atg7, a protein that is reputed to be essential for autophagy. These results establish novel functional links between microtubules and autophagy, identify a new chemotherapy resistance-associated autophagic defect, and describe the existence of LC3 puncta outside from autophagosomes. PMID- 20023421 TI - DNA double-strand breaks and ATM activation by transcription-blocking DNA lesions. AB - A taxia telangiectasia mutated (ATM), the deficiency of which causes a severe neurodegenerative disease, is a crucial mediator for the DNA double-strand break (DSB) response. We recently showed that transcription-blocking topoisomerase I cleavage complexes (TOP1cc) produce DSBs related to R-loop formation and activate ATM in post-mitotic neurons and lymphocytes. Here we discuss how TOP1cc can produce transcription arrest with R-loop formation and generate DSBs that activate ATM, as well as data suggesting that those transcription-dependent DSBs tend to form at the IgH locus and at specific genomic sites. We also address the potential roles of ATM in response to transcription-blocking TOP1cc. PMID- 20023422 TI - Enhancing perifosine's anticancer efficacy by preventing autophagy. AB - Our long-term research goal is to develop efficacious regimens for cancer therapy through our understanding of cancer biology and drug mechanisms. Perifosine is an alkylphospholipid exhibiting antitumor activity and is currently being tested in clinical trials. Its activity is partly associated with its ability to inhibit Akt activity. In an effort to understand the mechanism by which perifosine exerts its anticancer activity, our recent work shows that perifosine, in addition to inhibition of Akt, inhibits mTOR signaling through a different mechanism than classical mTOR inhibitors such as rapamycin via facilitating the degradation of major components in the mTOR axis including mTOR, raptor and rictor. Accordingly, perifosine substantially induces autophagy in addition to apoptosis. The combination of perifosine with a lysosomal inhibitor enhances apoptosis and inhibition of the growth of xenografts in nude mice, suggesting that perifosine induced autophagy protects cells from undergoing apoptosis. Thus, our findings highlight a novel mechanism accounting for perifosine's anticancer activity involving degradation-mediated mTOR inhibition and also suggest a potential strategy to enhance perifosine's anticancer efficacy by preventing autophagy. PMID- 20023423 TI - H2A.Z overexpression promotes cellular proliferation of breast cancer cells. AB - We recently showed that histone H2A.Z, as well as members of the ATP-dependent p400 chromatin remodeling complex (p400.com), are essential components of estrogen receptor alpha (ERalpha) signaling. More specifically, we showed that H2A.Z and p400.com are incorporated into the promoter regions of ERalpha target genes only upon gene induction, and also in a cyclic fashion. RNAi-mediated cellular depletion of H2A.Z and p400.com strongly impedes estrogen-dependent growth of breast cancer cells as well as strongly affect ERalpha-target gene expression. Two mechanisms emerged from our studies of how H2A.Z incorporation within ERalpha-target regulatory regions can actually regulate estrogen-mediated signaling: (1) by stabilizing nucleosomes within the translational DNA axis, a process that allows general transcription factors to be efficiently recruited to promoter regions; (2) by allowing estrogen-responsive enhancer function. In the current study, we now show that in MCF7 cells, ectopic overexpression of H2A.Z increases proliferation, and such in conditions where estrogen levels are low. Also, immunohistochemical studies of breast cancer biopsies show that the presence of H2A.Z correlates highly with that of ERalpha, but is associated with high-grade ER-negative cancers. Finally we show that ERalpha directly associates to the H2A.Z promoter, and consequently modulates its expression. Our study provides a possible link between H2A.Z and endocrine resistance by showing that H2A.Z overexpression leads to increased growth, particularly when estrogen levels are very low. PMID- 20023424 TI - Role of cytokinin responsive two-component system in ABA and osmotic stress signalings. PMID- 20023425 TI - Identification and prediction of abiotic stress responsive transcription factors involved in abiotic stress signaling in soybean. AB - Abiotic stresses such as extreme temperature, drought, high salinity, cold and waterlogging often result in significant losses to the yields of economically important crops such as soybean (Glycine max L.). Transcription factors (TFs) which bind to DNA through specific cis-regulatory sequences either activate or repress gene transcription have been reported to act as control switches in stress signaling. Recent completion of the soybean genomic sequence has open wide opportunities for large-scale identification and annotations of regulatory TFs in soybean for functional studies. Within the soybean genome, we identified 5,035 TF models which grouped into 61 families. Detailed annotations of soybean TF genes can be accessed at SoybeanTFDB (soybeantfdb.psc.riken.jp). Moreover, we have reported a new idea of high throughput prediction and selection of abiotic stress responsive TFs based on the existence of known stress responsive cis-element(s) located in the promoter regions of respective TFs and GO annotations. We, therefore, have provided a basic platform for the genome-wide analysis of regulatory mechanisms underlying abiotic stress responses and a reliable tool for prediction and selection of stress responsive TFs for further functional studies and genetic engineering. PMID- 20023426 TI - High-throughput transient transformation of Arabidopsis roots enables systematic colocalization analysis of GFP-tagged proteins. AB - Determination of the subcellular localization of an unknown protein is a major step towards the elucidation of its function. Lately, the expression of proteins fused to fluorescent markers has been very popular and many approaches have been proposed to express these proteins. Stable transformation using Agrobacterium tumefaciens generates stable lines for downstream experiments, but is time consuming. If only colocalization is required, transient techniques save time and effort. Several methods for transient assays have been described including protoplast transfection, biolistic bombardment, Agrobacterium tumefaciens cocultivation and infiltration. In general colocalizations are preferentially performed in intact tissues of the same species, resembling the native situation. High transformation rates were described for cotyledons of Arabidopsis, but never for roots. Here we report that it is possible to transform Arabidopsis root epidermal cells with an efficiency that is sufficient for colocalization purposes. PMID- 20023427 TI - A role for Chk2 in DNA damage induced mitotic delays in human colorectal cancer cells. AB - Progression into mitosis in the presence of DNA damage leads to spindle checkpoint (SAC) dependent mitotic delays and cytokinesis failure. In Drosophila embryos, DNA damage does not delay mitotic entry but triggers Checkpoint kinase-2 (Chk2) kinase dependent delays in mitotic exit. It is unclear if damage associated mitotic delays in human cells result from kinase signaling or breaks in centromere DNA that disrupt kinetochore function and activate the SAC. We show that transgenic expression of Human Chk2 in a Drosophila chk2 mutant background restores damage induced mitotic delays during early embryogenesis. Parental HCT116 colorectal cancer cells that progress into mitosis following DNA damage, due to either G(2) checkpoint adaptation or G(2) checkpoint abrogation by caffeine or the Chk1 inhibitor UCN-01, delay in mitosis and show high rates of cytokinesis failure. Significantly, these mitotic responses are suppressed in HCT116 chk2 knockout cells, and the response is restored by transgenic expression of wild type Chk2. However, both parental and chk2(-/-)HCT116 cells arrested in G(2) for prolonged periods by DNA damage prior to release from the G(2) block do show significant mitotic delays. Chk2 thus appears to have a conserved function in control of mitotic progression following G(2)/M transition with DNA damage. However, prolonged G(2) arrest with DNA damage can trigger Chk2 independent mitotic delays that may be secondary to kinetochore disruption. PMID- 20023428 TI - Beclin 1 modulates the anti-apoptotic activity of Bcl-2: insights from a pathogen infection system. AB - Coxiella burnetii is an obligate intracellular bacterium that generates large vacuoles in which this pathogen replicates and survives. We have previously demonstrated that C. burnetii interacts with the autophagic pathway as a strategy for its survival and replication. Coxiella displays an anti-apoptotic activity to maintain host cell viability, leading to a persistent infection. Our recent study reveals that Beclin 1 is recruited to the Coxiella-membrane vacuole favoring its development and bacterial replication. In contrast, the anti-apoptotic protein Bcl-2 alters the normal development of the Coxiella-replicative compartment. In addition, our results indicate that C. burnetii infection modulates autophagy and apoptotic pathways via Beclin 1-Bcl-2 interplay to establish a successful infection in the host cell. Of note, this pathogen-host cell model has allowed uncovering a novel function of Beclin 1 as a regulator of the anti-apoptotic activity of Bcl-2. We have also established that a proper interplay between Beclin 1 and Bcl-2 is required for both autophagy and apoptosis modulation. PMID- 20023429 TI - Synergy and antagonism of macroautophagy and chaperone-mediated autophagy in a cell model of pathological tau aggregation. AB - Tau aggregation characterizes a series of neurodegenerative diseases including AD and other tauopathies. The distribution of Tau deposits correlates with the loss of neurons in these neurodegenerative diseases, and Tau-induced toxicity depends on its ability to aggregate. We have used an inducible cell model to study the expression of Tau variants, the buildup of aggregates, and their removal by the autophagy-lysosomal system. Incomplete chaperone-mediated autophagy of Tau generates amyloidogenic fragments that promote aggregation. The Tau aggregates are removed from cells by macroautophagy. Thus the two autophagic pathways could become possible therapeutic targets. PMID- 20023430 TI - RFC3 regulates cell proliferation and pathogen resistance in Arabidopsis. PMID- 20023431 TI - Plant autophagy puts the brakes on cell death by controlling salicylic acid signaling. AB - It has long been recognized that autophagy in plants is important for nutrient recycling and plays a critical role in the ability of plants to adapt to environmental extremes such as nutrient deprivation. Recent reverse genetic studies, however, hint at other roles for autophagy, showing that autophagy defects in higher plants result in early senescence and excessive immunity related programmed cell death (PCD), irrespective of nutrient conditions. Until now, the mechanisms by which cells die in the absence of autophagy were unclear. In our study, using biochemical, pharmacological and genetic approaches, we reveal that excessive salicylic acid (SA) signaling is a major factor in autophagy-defective plant-dependent cell death and that the SA signal can induce autophagy. These findings suggest a novel physiological function for plant autophagy that operates via a negative feedback loop to modulate proper SA signaling. PMID- 20023432 TI - Biological evaluation of paclitaxel-peptide conjugates as a model for MMP2 targeted drug delivery. AB - Paclitaxel (PTX) is a highly effective cytotoxic agent widely used for the treatment of several solid tumors. However, PTX shows dose-limiting cytotoxicity and in most cases induces drug resistance followed by failure in treatment. To enhance the therapeutic index of a given drug, various drug delivery methods have been explored to systemically deliver sufficient amount of the drug to the desired site. In the present study, we designed and synthesized two PTX prodrugs by conjugating PTX at different sites with an octapeptide (AcGPLGIAGQ) that can be cleaved by MMP2 at tumor sites. As a result, PTX is expected to be released at the tumor sites, absorbed by the tumor cells, and thereby inhibit the tumor growth. We evaluated the in vitro activities of the two drugs in a panel of drug sensitive and -resistant cancer cell lines and their in vivo efficacies in a HT1080 fibrosarcoma mouse xenograft model that overexpresses MMP2. Our in vitro results showed that the PTX-AcGPLGIAGQ conjugates inhibited cancer cell proliferation with higher activity compared to that observed for free PTX, both of which were mediated by an arrest of G(2)/M-phase of the cell cycle. Consistent with the in vitro results, treatment with PTX-octapeptide conjugate resulted in extensive areas of necrosis and a lower percentage of proliferating cells in xenograft tumor sections. Together, our results indicate the potential of the tumor-targeted delivery of PTX to exploit the specific recognition of MMP2, reduce toxicity, and selectively kill tumor cells. PMID- 20023433 TI - ZIP4 upregulates the expression of neuropilin-1, vascular endothelial growth factor, and matrix metalloproteases in pancreatic cancer cell lines and xenografts. AB - We have recently found that a zinc transporter, ZIP4, is overexpressed in human pancreatic cancer and contributes to pancreatic cancer pathogenesis and progression. However, the detailed mechanism that how ZIP4 regulates pancreatic cancer growth is not clear. In this study, we further investigated the key molecules regulated by ZIP4 in pancreatic cancer angiogenesis and metastasis. We found that overexpression of ZIP4 caused significantly increased expression of NRP-1, VEGF, MMP-2 and MMP-9 in both pancreatic cancer cell lines and xenografts. Conversely, silencing of ZIP4 by short hairpin RNA (shRNA) was associated with decreased expression of NRP-1 and VEGF in pancreatic cancer xenografts. The ZIP4 expression and NRP-1 level are also correlated in established human pancreatic cancer cell lines. These results indicate that ZIP4-mediated pancreatic cancer growth might involve angiogenesis, invasion and metastasis pathways, and NRP-1, VEGF and MMPs are important intermediate molecules in transducing the ZIP4 initiated signal cascades in pancreatic cancer. PMID- 20023434 TI - pRB and E2F4 play distinct cell-intrinsic roles in fetal erythropoiesis. AB - The retinoblastoma tumor suppressor protein pRB functions, at least in part, by directly binding to and modulating the activity of the E2F transcription factors. Previous studies have shown that both E2F4 and pRB play important roles in fetal erythropoiesis. Given that these two proteins interact directly we investigated the overlap of E2F4 and pRB function in this process by analyzing E2f4(-/-), conditional Rb knockout (Rb(1lox/1lox)), and compound E2f4(-/-);Rb(1lox/1lox) embryos. At E15.5 E2f4(-/-) and Rb(1lox/1lox) fetal erythroid cells display distinct abnormalities in their differentiation profiles. When cultured in vitro, both E2f4(-/-) and Rb(1lox/1lox) erythroid cells show defects in cell cycle progression. Surprisingly, analysis of cell cycle profiling suggests that E2F4 and pRB control cell cycle exit through different mechanisms. Moreover, only pRB, but not E2F4, promotes cell survival in erythroid cells. We observed an additive rather than a synergistic impact upon the erythroid defects in the compound E2f4( /-);Rb(1lox/1lox) embryos. We further found that fetal liver macrophage development is largely normal regardless of genotype. Taken together, our results show that E2F4 and pRB play independent cell-intrinsic roles in fetal erythropoiesis. PMID- 20023435 TI - Learning and doing: operational research and access to HIV treatment in Africa. PMID- 20023436 TI - Higher risk of unsafe sex and impaired quality of life among patients not receiving antiretroviral therapy in Cameroon: results from the EVAL survey (ANRS 12-116). AB - OBJECTIVE: Cameroon has initiated a national programme of HIV care decentralization providing access to antiretroviral therapy (ART) for patients with CD4 cell counts less than 200 cells/microl or AIDS stage. Current clinical research suggests these criteria may be too stringent. This study aimed at evaluating the effect of not receiving ART on patients' psychosocial outcomes. DESIGN: The national cross-sectional survey EVAL (ANRS 12-116) collected psychosocial and clinical data for 3151 patients attending HIV services (September 2006 to March 2007). METHODS: Propensity score matching was used to control for demographic/clinical-immunological differences between patients receiving ART and those who did not. Generalized linear models were used to assess the impact, for different CD4 cell levels, of "not receiving" ART on health-related quality of life (HRQoL) inconsistent condom use with a sexual partner either serodiscordant or of unknown HIV status, self-reported symptoms and disclosure of HIV status to relatives or friends. RESULTS: Seventy-eight per cent of patients included in the survey were receiving ART. Non-treated patient breakdown was as follows: 8% (CD4<200 or AIDS stage), 5% (200350). In the multivariate matched-pairs analysis, impaired physical HRQoL, more frequent inconsistent condom use, more self-reported symptoms and less frequent disclosure of HIV status were all significantly associated (P < 0.0001) with not receiving ART, irrespective of the CD4 cell level. CONCLUSION: In addition to increasing clinical effectiveness, earlier initiation of ART at less severe immune-depression levels than previously recommended by World Health Organization guidelines for low-resource settings may be justified for improving subjective health and positive prevention among people living with HIV. PMID- 20023437 TI - Non-nucleoside reverse transcriptase inhibitor outcomes among combination antiretroviral therapy-treated adults in Botswana. AB - BACKGROUND: National initiatives offering non-nucleoside reverse transcriptase inhibitor (NNRTI)-based combination antiretroviral therapy (cART) have expanded in sub-Saharan Africa. The Tshepo study is the first clinical trial evaluating the long-term efficacy and tolerability of efavirenz versus nevirapine-based cART among adults in Botswana. METHODS: A 3-year randomized study (n = 650) using a 3 x 2 x 2 factorial design comparing efficacy and tolerability among: (i) zidovudine/lamivudine versus zidovudine/didanosine versus stavudine/lamivudine; (ii) efavirenz versus nevirapine; and (iii) community-based supervision versus standard adherence strategies. This paper focuses on comparison (ii). RESULTS: There was no significant difference by assigned NNRTI in time to virological failure with resistance (log-rank P = 0.14), nevirapine versus efavirenz [risk ratio (RR) 1.54, 95% CI 0.86-2.70]. Rates of virological failure with resistance were 9.6% nevirapine-treated (95% CI 6.8-13.5) versus 6.6% efavirenz-treated (95% CI 4.2-10.0) at 3 years. Women receiving nevirapine-based cART trended towards higher virological failure rates when compared with efavirenz-treated women, Holm corrected (log-rank P = 0.072), nevirapine versus efavirenz (RR 2.22, 95% CI 0.94 5.00). A total of 139 patients had 176 treatment-modifying toxicities, with a shorter time to event in nevirapine-treated versus efavirenz-treated patients (RR 1.85, 1.20-2.86; log-rank P = 0.0002). CONCLUSION: Tshepo-treated patients had excellent overall immunological and virological outcomes, and no significant differences were observed by randomized NNRTI comparison. Nevirapine-treated women trended towards higher virological failure with resistance compared with efavirenz-treated women. Nevirapine-treated adults had higher treatment modifying toxicity rates when compared with those receiving efavirenz. Nevirapine-based cART can continue to be offered to women in sub-Saharan Africa if patient education concerning toxicity is emphasized, routine safety monitoring chemistries are performed and the potential risk of efavirenz-related teratogenicity is considered. PMID- 20023438 TI - Who starts antiretroviral therapy in Durban, South Africa?... not everyone who should. AB - OBJECTIVE: To evaluate rates of antiretroviral therapy (ART) initiation within 12 months of a new HIV diagnosis in Durban, South Africa. DESIGN: Prospective observational cohort. METHODS: Adults (>or=18 years) were enrolled before HIV testing at two outpatient clinics into the South African Test, Identify and Link cohort. Both sites offer comprehensive HIV care. HIV test results, CD4 cell counts, dates of ART initiation and dates of death were collected from medical records and 12-month patient/family interviews were conducted. ART eligibility was defined as a CD4 cell count less than 200 cells/microl within 90 days of HIV diagnosis. The primary endpoint was ART initiation within 12 months for ART eligible subjects. RESULTS: From November 2006 to October 2008, 1474 newly diagnosed HIV-infected outpatients were enrolled, 1012 (69%) of whom underwent CD4 cell count testing within 90 days. The median CD4 cell count was 159 cells/microl (interquartile range 65-299). Of those who underwent CD4 cell count testing, 538 (53%) were ART-eligible. Only 210 (39%) eligible enrollees were known to have initiated ART within 12 months. Among ART-eligible subjects, there were 108 known deaths; 82% occurred before ART initiation or with unknown ART initiation status. Men [rate ratio (RR) 1.3, 95% confidence interval (CI) 1.1 1.5] and subjects without an HIV-infected family member/friend (RR 1.3, 95% CI 1.1-1.7) were more likely not to start ART. CONCLUSION: Less than half of ART eligible subjects started ART within 12 months. Substantial attrition and mortality follow HIV diagnosis before ART initiation in Durban, South Africa. Major efforts directed towards earlier HIV diagnosis, effective linkage to care and timely ART initiation are urgently needed. PMID- 20023439 TI - Can the deployment of community health workers for the delivery of HIV services represent an effective and sustainable response to health workforce shortages? Results of a multicountry study. AB - In countries severely affected by HIV/AIDS, shortages of health workers present a major obstacle to scaling up HIV services. Adopting a task shifting approach for the deployment of community health workers (CHWs) represents one strategy for rapid expansion of the health workforce. This study aimed to evaluate the contribution of CHWs with a focus on identifying the critical elements of an enabling environment that can ensure they provide quality services in a manner that is sustainable. The method of work included a collection of primary data in five countries: Brazil, Ethiopia, Malawi, Namibia, and Uganda. The findings show that delegation of specific tasks to cadres of CHWs with limited training can increase access to HIV services, particularly in rural areas and among underserved communities, and can improve the quality of care for HIV. There is also evidence that CHWs can make a significant contribution to the delivery of a wide range of other health services. The findings also show that certain conditions must be observed if CHWs are to contribute to well-functioning and sustainable service delivery. These conditions involve adequate systems integration with significant attention to: political will and commitment; collaborative planning; definition of scope of practice; selection and educational requirements; registration, licensure and certification; recruitment and deployment; adequate and sustainable remuneration; mentoring and supervision including referral system; career path and continuous education; performance evaluation; supply of equipment and commodities. The study concludes that, where there is the necessary support, the potential contribution of CHWs can be optimized and represents a valuable addition to the urgent expansion of human resources for health, and to universal coverage of HIV services. PMID- 20023440 TI - Scaling up access to antiretroviral treatment for HIV infection: the impact of decentralization of healthcare delivery in Cameroon. AB - BACKGROUND: The independent evaluation of the Cameroonian antiretroviral therapy (ART) Programme, which reached one of the highest coverage in the eligible HIV infected population (58%) in Sub-Saharan Africa, offered the opportunity to assess ART outcomes in the context of the decentralization of HIV care delivery. MATERIALS AND METHODS: A cross-sectional survey (EVAL, ANRS 12-116, 2007) was carried out in a random sample of 3151 HIV-positive patients (response rate 90%) attending 27 treatment centres at the different level of the healthcare delivery (central, provincial and district), as well as in the exhaustive sample of doctors in charge of HIV care in these centres (response rate 92%, n = 97). Multivariate two-level analyses were conducted to assess the impact of the level of healthcare delivery on CD4 cell gains since initiation of treatment and adherence to treatment in the subsample of patients who were ART-treated for 6 months or more (n = 1985). RESULTS: District treatment centres were characterized by more limited technical and human resources but a lower workload. ART-treated patients followed up in these centres had significantly lower socioeconomic status. After adjustment for other explanatory factors, immunological improvement was similar in patients followed up at the central and district level, whereas adherence to ART was better both at provincial and district levels. CONCLUSION: Success in scaling-up access to ART in Cameroon has been facilitated by decentralization of the healthcare system. Long-term sustainability urgently implies better integration of this HIV-targeted programme in the global healthcare reform of financing mechanisms, management of human resources and drug procurement systems. PMID- 20023441 TI - Quality of HIV care provided by non-physician clinicians and physicians in Mozambique: a retrospective cohort study. AB - OBJECTIVES: To compare HIV care quality provided by non-physician clinicians (NPC) and physicians. DESIGN: Retrospective cohort study assessing the relationship between provider cadre and HIV care quality among non-pregnant adult patients initiating antiretroviral therapy (ART) in the national HIV care programme. METHODS: Computerized medical records from patients initiating ART between July 2004 and October 2007 at two HIV public HIV clinics in central Mozambique were used to develop multivariate analyses evaluating differences in process and care continuity measures for patients whose initial provider was a NPC or physician. RESULTS: A total of 5892 patients was included in the study, including 4093 (69.5%) with NPC and 1799 (30.5%) with physicians as initial providers. Those whose initial provider was a NPC were more likely to have a CD4 cell count 90-210 days [risk ratio (RR) 1.13, 1.044 mm Hg on average and >5 mm Hg in >10% of recording time), 101 (93%) were responders (ie, increase in the NPH score of >2). Correspondingly, only 2 of 21 (10%) without increased ICP pulsatility were responders. Superficial wound infection was the only complication of ICP monitoring and occurred in 4 (2%) patients. CONCLUSION: Surgical results in iNPH were good with almost 80% of patients improving after treatment. The data indicate that improvement after surgery can be anticipated in 9 of 10 iNPH patients with abnormal ICP pulsatility, but in only 1 of 10 with normal ICP pulsatility. Diagnostic ICP monitoring had a low complication rate. PMID- 20023541 TI - Malignant tumors of the anterolateral skull base. AB - OBJECTIVE: Malignancies of the anterolateral skull base are clinically and pathologically distinct from those of the central anterior skull base and the temporal bone. The purpose of this report is to describe the outcomes and complications after skull base surgery and multimodality therapy in a group of patients with anterolateral skull base malignancies. PATIENT DATA AND METHODS: The mean duration of follow-up for living patients was 57.2 months (median, 56.8 months). The median age of the 52 patients who met the inclusion criteria for this study was 47 years (range, 1-81 years). The most common presenting feature was cranial nerve palsy (60%). Of these cranial nerve palsies, trigeminal neuropathies causing facial numbness were the most common, with V2 being affected in 35%, V3 affected in 33%, and V1 affected in 17%. Abducens neuropathy was present in 14% of patients. The most frequently occurring pathologies after the various sarcomas were squamous cell carcinoma (SCC) and adenoid cystic carcinoma (ACC) in 23% and 14% of patients, respectively. Of the 30 sarcomas, 16 were classified as low grade and 14 were classified as high grade. RESULTS: Complications of treatments were identified in 16 patients (31%). Ten patients had a single complication, whereas 6 patients experienced multiple complications. The most common complications were a new or worsened cranial nerve deficit (n = 4), pneumonia (n = 4), and flap necrosis (n = 3). Recurrence after the treatment associated with the index surgery occurred in 37 patients (71%). The recurrence was local in 30 patients (58%), both local and distant (metastatic) in 4 patients (8%), and only distant in 3 patients (12%). The median progression-free survival (PFS) was 2.1 years (range, 1.2-3.0 years). Median PFS times of 0.6 and 1.6 years were noted for patients with high-grade sarcoma (HGS) and low-grade sarcoma (LGS), respectively. The mean PFS (median not reached) for the patients with SCC was 4.6 years, whereas the median PFS for patients with ACC was 3.3 years. The overall 2- and 5-year survivals for all patients were 81% and 53% (median, 5.0 years; 95% confidence interval, 3.9-6.1 years), respectively. The median survival for patients with nonsarcomas was 6.9 years, the 2-year survival was 82%, and the 5-year survival was 55%. Patients with HGS survived the shortest time (median, 3.3 years; 2-year, 64%; 5-year, 27%), whereas those patients with LGS had an intermediate survival (median, 5.3 years; 2-year, 94%, 5-year, 72%). CONCLUSION: It is our belief that anterolateral skull base malignancies comprise a distinct group of tumors. These lesions should be analyzed separately from central anterior skull base lesions and temporal bone malignancies. With a multimodality treatment protocol, acceptable survivals may be obtained that are comparable to results that have been reported for tumors involving less difficult areas of the skull base. PMID- 20023542 TI - Endovascular treatment increases but gamma knife radiosurgery decreases angiogenic activity of arteriovenous malformations: an in vivo experimental study using a rat cornea model. AB - OBJECTIVE: To compare the angiogenic potentials of embolized, gamma knife-treated or untreated cerebral arteriovenous malformations (AVMs), using a rat cornea angiogenesis model. METHODS: Tissue samples from cerebral AVM patients who were either untreated or had previously been treated with embolization or gamma knife radiosurgery and who had undergone operations for hemorrhage at the Neurosurgery Department or the Neurological Sciences Institute of Marmara University were used. For the macroscopic evaluation of angiogenesis, tissue samples were inoculated in a micropocket created on the rat eye, and the level of angiogenic activity was graded macroscopically for 15 days, with glioblastoma multiforme and normal brain artery tissues serving as positive and negative controls, respectively. For the other part of the experiment, eyes of another set of rats were inoculated with the study samples only using the same cornea angiogenesis model, in which microvessel count and vascular endothelial growth factor assessment was done at days 3, 7, 11, and 15. RESULTS: Based on our macroscopic findings in the cornea angiogenesis model, embolized AVMs exhibited the highest angiogenic activity, followed by untreated AVMs and gamma knife-treated AVMs. Evaluations of vascular endothelial growth factor expression and microvessel counts showed a similar relation among the 3 tissue groups with regard to the level of angiogenic activity, supporting the results of macroscopic examinations. CONCLUSION: This study, for the first time, provides experimental semiquantitative data to compare the angiogenic potentials of embolized and gamma knife-treated AVM tissues. Embolization may increase angiogenic activity, and gamma knife radiosurgery may decrease it when compared with activity in previously untreated AVMs. These data can be useful to understand why recurrence of AVMs after angiographically demonstrated endovascular occlusion is common but after gamma knife occlusion is rare. PMID- 20023543 TI - Manganese-enhanced magnetic resonance imaging in experimental spinal cord injury: correlation between T1-weighted changes and Mn(2+) concentrations. AB - OBJECTIVE: Manganese (Mn(2+))-enhanced magnetic resonance imaging (MEMRI) is a potentially important tool for assessing neural tissue regeneration after spinal cord injury (SCI). We evaluated the relation between Mn(2+) and T1-weighted magnetic resonance (MR) signals in an SCI rat model. METHODS: Rats were divided into 4 groups with or without SCI (T9-level transection) and with or without Mn(2+) injection. Two microliters of 0.2 mol/L MnCl(2) was injected into the lateral ventricles. Magnetic resonance imaging (MRI) was performed 60 hours after injection. Signal intensities at cervical, thoracic, and lumbar levels were measured and normalized to the intensity of perivertebral muscles. Spinal cord sections were analyzed by inductively coupled plasma mass spectrometry (ICP-MS) for total Mn(2+) content. The results of ICP-MS were compared with MR signal intensity. RESULTS: T1-weighted MR signal intensity and ICP-MS-measured Mn(2+) were significantly decreased below the SCI injury site in Mn(2+)-injected groups with or without SCI, and were similar to intensity and Mn(2+) levels of noninjected animals. Signal intensity and Mn(2+) concentration tended to decrease from cervical to lumbar spinal levels in the control rats. ICP-MS data correlated with MRI results. CONCLUSION: The results confirmed Mn(2+) uptake in the spinal cord after intraventricular injection. T1-weighted MR signal intensity correlates with spinal Mn(2+) concentration as measured with ICP-MS. This work establishes the repeatability of MEMRI of the injured spinal cord and makes it possible to compare changes in axonal transport rates through the spinal cord after neuronal regeneration in vivo at different stages. MEMRI in animal models may improve understanding of the factors required to promote spinal cord regeneration. PMID- 20023544 TI - An automated algorithm to improve the precision of basilar artery diameter measurements before and after subarachnoid hemorrhage-induced vasospasm in an animal model. AB - OBJECTIVE: Quantifying vasospasm has traditionally been performed manually, a method prone to imprecision and user bias. An alternative approach is to use computerized image analysis techniques to define and quantify the diameter of a vessel. The goal of this article is to demonstrate a novel automated vessel measurement algorithm specific to the needs of vasospasm studies and to compare it with traditional manual measurements in an animal model of vasospasm. METHODS: A total of 576 arterial diameter measurements were collected by 4 independent, blinded examiners from 24 angiograms in a rabbit subarachnoid hemorrhage (SAH) model. Measurements were taken from 3 segments of the basilar artery in anteroposterior and lateral projections, both before SAH and after SAH-induced vasospasm. Means and standard deviations of 288 manual measurements were compared with 288 automated measurements. RESULTS: The precision of automated measurements was significantly improved compared with standardized manual measurements (85.7% decrease in variation; P < .001). When using automated measurements, the precision was not affected by vessel size, but when using manual measurements, smaller arteries were less precise (P = .04). There was no significant difference in precision between 2 different contrast concentrations (P = .32). CONCLUSION: Automated measurements of basilar artery diameters are more precise than manual measurements, both before and after SAH-induced vasospasm. The variability in the manual group worsens when the artery is smaller secondary to vasospasm, indicating a need for the use of this segmentation method. PMID- 20023545 TI - Platelet-derived growth factor BB mediates the tropism of human mesenchymal stem cells for malignant gliomas. AB - OBJECTIVE: Bone marrow-derived human mesenchymal stem cells (hMSCs) are capable of localizing to gliomas after systemic delivery and can be used in glioma therapy. However, the mechanism underlying the tropism of hMSCs for gliomas remains unclear. In vitro studies suggest that platelet-derived growth factor BB (PDGF-BB) may mediate this tropism. However, a causal role of PDGF-BB has not been demonstrated in vivo. Therefore, we tested the hypothesis that PDGF-BB mediates the attraction of hMSCs to gliomas in vitro and in vivo. METHODS: U87 or LN229 glioma cells were transfected with plasmids encoding human PDGF-B. Stable transfected clones that secreted large amounts of PDFG-BB and clones that produced low levels of PDGF were chosen. In vitro migration of hMSCs toward PDGF B or conditioned media from high- and low-secreting PDGF-B tumor cells was assessed using Matrigel invasion assays. For in vivo localization studies, hMSCs were tracked by bioluminescence imaging (BLI) after transduction with an adenovirus containing luciferase cDNA. In other studies, hMSCs were labeled with green fluorescent protein (gfp) and analyzed for intratumoral localization by immunohistochemistry. RESULTS: In vitro invasion assays showed that significantly more hMSCs migrated toward glioma cells engineered to secrete high levels of PDGF BB compared with low-secreting gliomas. Anti-PDGF-BB-neutralizing antibody abrogated this increase in migration. Pretreatment of hMSCs with inhibitory antibodies against PDGF receptor-beta also reduced hMSC migration. To demonstrate that PDGF-BB mediates the localization of hMSCs in vivo, hMSCs-Ad-Luc were injected into the carotid artery of mice harboring orthotopic 7-day-old U87-PDGF BB-high secreting or U87-PDGF-BB-low secreting xenografts and analyzed by BLI. Statistically significant increases in hMSCs were seen within PDGF-BB-high xenografts compared with PDGF-BB-low xenografts. To control for PDGF-BB-induced differences in tumor size and vascularity, gfp-labeled hMSCs were injected into the carotid arteries of animals harboring 4-day old PDGF-BB-high secreting xenografts or 7-day old PDGF-BB-low secreting xenografts. At these times tumors had similar size and vessel density. Statistically significant more hMSCs localized to PDGF-BB-high secreting xenografts compared with PDGF-BB-low secreting xenografts. Pretreatment of hMSCs with anti-PDGFR-beta-inhibitory antibodies decreased the localization of hMSCs in this intracranial model. CONCLUSION: PDGF-BB increases the attraction of hMSCs for gliomas in vitro and in vivo, and this tropism is mediated via PDGF-beta receptors on hMSCs. These findings can be exploited for advancing hMSC treatment. PMID- 20023546 TI - Effect of vasodilation by milrinone, a phosphodiesterase III inhibitor, on vasospastic arteries after a subarachnoid hemorrhage in vitro and in vivo: effectiveness of cisternal injection of milrinone. AB - OBJECTIVE: Cerebral vasospasm remains a major cause of morbidity and mortality. Milrinone, a bipyridine phosphodiesterase III inhibitor, is a potent member of the inodilator class of cardiac agents for vasospasm and is injected intra arterially or intracisternally. There have been no studies investigating the duration of action (context-sensitive half-life) of milrinone for vasospasm or the most effective route of administration (intra-arterial versus intracisternal). We examined the effects of intracisternal and intra-arterial injection of milrinone on chronic cerebral vasospasm in dogs. METHODS: A double hemorrhage canine model was used. In a preliminary isometric tension study of canine vasospastic basilar arteries, the vasodilatory effects of milrinone were examined 7 days after an initial injection of blood. Milrinone was injected intracisternally (0.1 mg, 0.47 mmol/L) or intra-arterially (0.3 mg/kg, 1.2 mmol/L), and angiograms were performed 30, 60, 120, 180, 240, 300, and 360 minutes later on day 7. RESULTS: Milrinone produced concentration-dependent vasodilation and was effective intracisternally, resulting in significant dilation until 180 minutes after injection and a tendency for dilation until 240 minutes. The effect of intra-arterial injection was not as significant compared with an intracisternal injection, resulting in significant dilation only at 180 minutes after intra-arterial injection. CONCLUSION: Intracisternal injection of milrinone was more effective than intra-arterial injection for chronic cerebral vasospasm in dogs because intracisternal injection produced a higher concentration in vasospastic arteries (0.034-0.068 mmol/L intracisternally versus 0.016 mmol/L intra-arterially). PMID- 20023547 TI - Activated protein C is neuroprotective and mediates new blood vessel formation and neurogenesis after controlled cortical impact. AB - OBJECTIVE: Activated protein C (APC) is neuroprotective in stroke models and promotes postischemic neovascularization and neurogenesis. We used a controlled cortical impact (CCI) in mice to determine the effects of APC on neuroprotection and angiogenesis and neurogenesis after traumatic brain injury (TBI). METHODS: Mice were given (1) single-dose APC (0.8 mg/kg intraperitoneally) 15 minutes after injury, (2) multidose APC (0.8 mg/kg intraperitoneally) 15 minutes and 6 to 48 hours after injury, or (3) vehicle. We then assessed the effects of APC on posttraumatic motor function with the rotarod and wire grip and beam balance tasks, and we determined the lesion volumes and studied the formation of new blood vessels and markers of neurogenesis. RESULTS: Mice treated with single-dose or multidose APC, compared with vehicle, showed significantly improved motor function on all tests. In the single-dose and multidose APC treatment groups, at 7 days after treatment, lesion volume was significantly decreased by 30% and 50%, respectively. Multidose APC, but not single-dose APC, increased new blood vessel formation as shown by CD105(+)/Ki-67(+) double immunostaining by nearly 2-fold at 7 days. Multidose APC also promoted posttraumatic proliferation of neuroblasts in the subventricular zone (SVZ) and their migration from the SVZ to the perilesional area. CONCLUSION: Activated protein C improves functional outcome and is neuroprotective after TBI. It also promotes angiogenesis and survival and migration of neuroblasts from the SVZ to the perilesional area, but the exact role of these brain repair mechanisms remains to be determined. The present findings suggest that APC therapy may hold a significant therapeutic potential for TBI. PMID- 20023548 TI - Intrasellar ultrasound in transsphenoidal surgery: a novel technique. AB - OBJECTIVE: Residual tumor masses are common after transsphenoidal surgery. The risk of a residual mass increases with tumor size and parasellar or suprasellar growth. Transsphenoidal surgery is usually performed without image guidance. We aimed to investigate a new technical solution developed for intraoperative ultrasound imaging during transsphenoidal surgery, with respect to potential clinical use and the ability to identify neuroanatomy and tumor. METHODS: In 9 patients with pituitary macroadenomas, intrasphenoidal and intrasellar ultrasound was assessed during transsphenoidal operations. Ultrasound B-mode, power-Doppler and color-Doppler images were acquired using a small prototype linear array, side looking probe. The long probe tip measures only 3 x 4 mm. We present images and discuss the potential of intrasphenoidal and intrasellar and ultrasound in transsphenoidal surgery. RESULTS: We present 2-dimensional, high-resolution ultrasound images. A small side-looking, high-frequency ultrasound probe can be used to ensure orientation in the midline for the surgical approach to identify important neurovascular structures to be avoided during surgery and for resection control and identification of normal pituitary tissue. The image resolution is far better than what can be achieved with current clinical magnetic resonance imaging technology. CONCLUSION: We believe that the concept of intrasellar ultrasound can be further developed to become a flexible and useful tool in transsphenoidal surgery. PMID- 20023549 TI - Integrating magnetic resonance imaging postprocessing results into neuronavigation for electrode implantation and resection of subtle focal cortical dysplasia in previously cryptogenic epilepsy. AB - OBJECTIVE: Focal cortical dysplasias (FCDs) are highly epileptogenic lesions. Surgical removal is frequently the best treatment option for pharmacoresistant epilepsy. However, subtle FCDs may remain undetected even after high-resolution magnetic resonance imaging (MRI). Morphometric MRI analysis, which compares the individual brain with a normal database, can facilitate the detection of FCDs. We describe how the results of normal database-based MRI postprocessing can be used to guide stereotactic electrode implantation and subsequent resection of lesions that are suspected to be FCDs. METHODS: A presurgical evaluation was conducted on a 19-year-old woman with pharmacoresistant hypermotor seizures. Conventional high resolution MRI was classified as negative for epileptogenic lesions. However, morphometric analysis of the spatially normalized MRI revealed abnormal gyration and blurring of the gray-white matter junction, which was suggestive of a small and deeply seated FCD in the left frontal lobe. RESULTS: The brain region highlighted by morphometric analysis was marked as a region of interest, transferred back to the original dimension of the individual MRI, and imported into a neuronavigation system. This allowed the region of interest-targeted stereotactic implantation of 2 depth electrodes, by which seizure onset was confirmed in the lesion. The electrodes also guided the final resection, which rendered the patient seizure-free. The lesion was histologically classified as FCD Palmini and Luders IIB. CONCLUSION: Transferring normal database-based MRI postprocessing results into a neuronavigation system is a new and worthwhile extension of multimodal neuronavigation. The combination of resulting regions of interest with functional and anatomic data may facilitate planning of electrode implantation for invasive electroencephalographic recordings and the final resection of small or deeply seated FCDs. PMID- 20023550 TI - Endoscopic approach to the infratemporal fossa: anatomic study. AB - OBJECTIVE: Classic surgical exposures of the infratemporal fossa region, including the adjacent intracranial space, temporal bone, and sinonasal region, require the extensive exposure associated with the transcranial, transfacial, and transmandibular approaches with their inherent neurological and cosmetic morbidities. In this study, we evaluated the feasibility and exposure afforded by combining 2 endoscopic transmaxillary approaches, endonasal and Caldwell-Luc supplement, to the infratemporal fossa. METHODS: Endoscopic transmaxillary dissection was performed in 4 formalin-fixed cadaver heads (8 sides). We quantified the extent of exposure achieved within the pterygopalatine and infratemporal fossae after our initial dissection, which was endonasal with a medial antrostomy, and after addition of a Caldwell-Luc incision with an anterior antrostomy. Complementing this anatomic study, we report on a patient in whom this endoscopic transmaxillary approach combining the endonasal and Caldwell-Luc approaches was used for resection of a trigeminal schwannoma in the infratemporal fossa. RESULTS: The combination of these 2 endoscopic transmaxillary approaches enabled visualization of the entire region of the pterygopalatine fossa and anteromedial aspect of the infratemporal fossa. Additional posterolateral exposure of the infratemporal fossa requires significant traumatic traction on the nose. Addition of the Caldwell-Luc transmaxillary approach exposed the remainder of the infratemporal fossa, including the mandibular nerve and branches, middle meningeal artery, and even the distal cervical portion of the internal carotid artery. CONCLUSION: Endoscopic exposure of the infratemporal fossa is feasible. Using the combination of the endonasal and Caldwell-Luc approaches for direct transmaxillary access significantly extended exposure, allowing safe and effective resection of infratemporal fossa lesions. PMID- 20023551 TI - Utilization of fluorescein for identification and preservation of the facial nerve and semicircular canals for safe mastoidectomy: a proof of concept laboratory cadaveric study. AB - OBJECTIVE: Mastoidectomy can be a very challenging procedure for many reasons. The normal anatomy can be distorted because of inflammatory processes and tumors and recurrences. Avoiding injuries to the semicircular canals (SCCs) and facial canal is mandatory, and there is need to find a way to recognize the facial nerve and SCCs for safe performance of mastoidectomy. We describe, as a proof of concept, a novel technique to drill the mastoid while allowing the surgeon to recognize and avoid injuries to vital structures, in the cadaver. METHODS: Four fresh cadaveric heads (8 sides) were prepared by cannulating the major vessels at the level of the neck. After removal of the mastoid cortex, indocyanine green was injected in the vessels. The sigmoid sinus alongside the facial nerve and SCCs was skeletonized using the drilling guidance provided by the fluorescence. The mucosa covering the air cells of the mastoid is very well vascularized compared with the thick bone representing the outer layer of the SCCs and facial canal. Consequently, after the indocyanine green injection, the mucosa shines whereas the bone does not. The fluorescence guides the drilling displaying air cells that are safe to remove. RESULTS: Eight mastoidectomies were performed, resulting in optimal drilling with no injuries to the facial canal and SCCs. CONCLUSION: With this novel technique, it is possible to perfectly skeletonize the facial nerve and the SCCs in the cadaver. We think that this technique can be an adjunct in the armamentarium of trainees that are not familiar with the anatomy of the temporal bone and eventually of neurosurgeons facing lesions that require the removal of various degrees of the mastoid. PMID- 20023552 TI - Clinical and pathological characteristics of brain metastasis resected after failed radiosurgery. AB - OBJECTIVE: This study evaluates the tumor histopathology and clinical characteristics of patients who underwent resection of their brain metastasis after failed gamma knife radiosurgery. METHODS: This study was a retrospective review from a prospective database. A total of 1200 brain metastases in 912 patients were treated by gamma knife radiosurgery during a 7-year period. Fifteen patients (1.6% of patients, 1.2% of all brain metastases) underwent resective surgery for either presumed tumor progression (6 patients) or worsening neurological symptoms associated with increased mass effect (9 patients). Radiographic imaging, radiosurgical and surgical treatment parameters, histopathological findings, and long-term outcomes were reviewed for all patients. RESULTS: The mean age at the time of radiosurgery was 57 years (age range, 32-65 years). Initial pathological diagnoses included metastatic non-small cell lung carcinoma in 8 patients (53%), melanoma in 4 patients (27%), renal cell carcinoma in 2 patients (13%), and squamous cell carcinoma of the tongue in 1 patient (7%). The mean time interval between radiosurgery and surgical extirpation was 8.5 months (range, 3 weeks to 34 months). The mean treatment volume for the resected lesion at the time of radiosurgery was 4.4 cm(3) (range, 0.6-8.4 cm(3)). The mean dose to the tumor margin was 21Gy (range, 18-24 Gy). In addition to the 15 tumors that were eventually resected, a total of 32 other metastases were treated synchronously, with a 78% control rate. The mean volume immediately before surgery for the 15 resected lesions was 7.5 cm(3) (range, 3.8 10.2 cm(3)). Histological findings after radiosurgery varied from case to case and included viable tumor, necrotic tumor, vascular hyalinization, hemosiderin laden macrophages, reactive gliosis in surrounding brain tissue, and an elevated MIB-1 proliferation index in cases with viable tumor. The mean survival for patients in whom viable tumor was identified (9.4 months) was significantly lower than that of patients in whom only necrosis was seen (15.1 months; Fisher's exact test, P < 0.05). CONCLUSION: Radiation necrosis and tumor radioresistance are the most common causes precipitating a need for surgical resection after radiosurgery in patients with brain metastasis. PMID- 20023554 TI - Point of care documentation impact on the nurse-patient interaction. AB - Electronic medical record (EMR) point-of-care (POC) documentation in patients' rooms is a recent shift in technology use in hospitals. POC documentation reduces inefficiencies, decreases the probability of errors, promotes information transfer, and encourages the nurse to be at the bedside. However, EMR POC documentation has the potential to distract the nurse's attention away from the patient and compromise the nurse-patient interaction. PMID- 20023555 TI - From the editor. Creating a culture of nursing engagement. PMID- 20023556 TI - Guest editorial. PMID- 20023557 TI - Nine structures and leadership practices essential for a magnetic (healthy) work environment. AB - Improving clinical nurse work environments is a major challenge faced by nurse executives today. To meet this challenge, nurse leaders must implement the "right" structures and best leadership practices so that clinical nurses can engage in the work processes and relationships that are empirically linked to quality patient outcomes. What are these "right" structures and best leadership practices? Meta-analyses of 2 sets of publications were used to identify organizational structures and best leadership practices essential to a healthy work environment, that is, a work environment that enables them to engage in the work processes and relationships needed for quality patient care outcomes. The first set was 12 publications from 7 professional organizations/regulatory bodies that advocated forces, hallmarks, and standards for a healthy work environment. The second set was 18 publications from the Essentials of Magnetism structure identification studies, in which the aggregated results from 1300 interviews with staff nurse, manager, and physician "experts" were compared with the agency results. Broadening the categories and final aggregation yielded the 9 most important and influential structures essential to a quality work environment. Suggestions for implementing these structures are provided. PMID- 20023558 TI - The culture of nursing engagement: a historical perspective. AB - Work engagement is key to organizational efforts to retain nurses and mitigate future nursing shortages. In their dual role as caregivers and organization "keepers," nurses may already have the key to creating a culture of engagement. The characteristics and benefits of "engaging" work environments are captured in nursing professional practice models, as evidenced by a quarter century of research on Magnet hospitals and professional practice. An inflection point, providing an opportunity for transformational change in the nursing work environment, may be generated by a critical need for nurses and a call for healthcare delivery system redesign. This article explores this opportunity for change by (1) examining history and nursing's deep roots in professional practice and its journey as an evolving profession and (2) mapping the growth of hospitals and the advancement of nursing's role in the United States, in the context of organizational theory. Finally, it examines the relationship and alignment of goals between the Institute of Medicine's call for system redesign and professional practice model elements, as the potential infrastructure for change. Focusing on the past and today provides us with a beginning framework from which to move expeditiously toward creating a culture of nursing engagement now. PMID- 20023559 TI - Transformation of a nursing culture through actualization of a nursing professional practice model. AB - Leading and effecting meaningful change in a nursing division culture, such as the type required to achieve Magnet designation, entails senior nurse executives to be well-acquainted not only with the facts and figures of their business but also with the nuances, myths, and cultures that either enable or block a change from occurring. Expert nurse leaders embrace the story being told by data on dashboards and the quality outcomes achieved and look beyond those points of information out to the edges of their division. These nurse executives also seek to understand the pivotal, perhaps seemingly inconsequential things (notions, beliefs, cultural beliefs, and stories) that will block or tip a culture to change and achieve success. At the Hospital of the University of Pennsylvania (HUP), a Magnet-designated organization, the road to Magnet was not straightforward. Instead, the path was a winding, learning journey. Through authentic leadership and the conception and actualization of a professional practice model, the HUP Nursing Excellence in Professional Practice (HUP NEPP) model, Magnet designation was achieved and a nursing culture was transformed. PMID- 20023560 TI - Strengthening the voice of the clinical nurse: the design and implementation of a shared governance model. PMID- 20023561 TI - Collaboration to partnerships. AB - Partnerships are at the center of the Hospital of the University of Pennsylvania Nursing Excellence Professional Practice (HUP-NEPP) model. Through the use of collaboration, skilled communication, and respectful workplace, partnerships can be formed, leading ultimately to world-class patient care. At HUP, interdisciplinary partnerships are evidenced by the clinical nurses through shared governance. This article describes the components necessary to form successful partnerships. PMID- 20023562 TI - Improving the work environment through the use of research instruments: an example. AB - Over the past several years, although the quality of the work environment has improved for registered nurses, there is much that still needs to be done. However, much of the published research conducted in relation to the quality of work environments has focused on the more traditional components of the work environment. To assess the nontraditional components of a work environment, it is critical that reliable and valid instruments be utilized in order to ensure that the data gathered is accurate and truly reflective of what exists within the work environment. This article will describe one instrument, the Sieloff-King Assessment of Group Outcome Attainment within Organizations, that has demonstrated reliability and validity. The application of this instrument in the improvement of the health care environment will be discussed. PMID- 20023563 TI - Development of a reliable and valid organization-specific professional practice assessment tool. AB - PURPOSE: This article describes a study to devise an organization-specific professional practice model (PPM) assessment that reflects actual unit involvement. A secondary study goal is the development of a unit-based index that can be used to conduct comparative analyses in an efficient way. DESIGN: Each of the 5 elements of the organization's PPM was represented by 1 or more items on an author-developed instrument. The tool was structured so that item scores could be summed to achieve a single subscale for each PPM element and further aggregated into a total score. METHODS: The instrument was administered to a 40% random sample of all regularly scheduled, full- and part-time registered nurses in an academic, community Magnet hospital in 2003 and 2005. Descriptive statistics were calculated for items, subscales, and summary scores for each patient care unit and overall. A weighted, unit-based index was developed to reflect each unit's score on a scale of 100. FINDINGS: The 2003 assessment response rate was 51% (n = 200); the 2005 response rate was 48% (n = 193). Subscale scores and a total PPM score were calculated by summing the values of each individual item. Submissions enabled calculations of total scores by unit, mean scores by item, and the development of a unit-specific PPM index of performance. CONCLUSIONS: Beyond shared principles of empowerment, the specifics of each organization's PPM may differ in those key components of care delivery nurses are empowered to effect. Thus, fidelity to the organization-specific PPM is not well tested with generic decisional-involvement instruments. An organization-specific assessment such as this one can provide evidence of not only organizational PPM fidelity but a quantitative method to ensure that staff nurse decisional involvement is continuously evolving to an ever higher state. PMID- 20023564 TI - Role conflict, role ambiguity, and job satisfaction in nurse executives. AB - This article reports the results of a cross-sectional correlational survey design of nurse executives' (NEs') views on role conflict, role ambiguity, job satisfaction, and depression. A total of 380 surveys were returned from 1000 randomly selected participants. Four instruments were utilized, 1 for demographic data and 3 established instruments. Results indicated that the respondents had low to moderate amounts of role ambiguity, high levels of job satisfaction, low levels of depression, a negative relationship between role conflict and role ambiguity and job satisfaction, and a positive relationship between role conflict and depression. Furthermore, although levels of role conflict are decreasing, levels of role ambiguity are increasing. Neither educational level nor age was related to the dependent variables. Role conflict and role ambiguity are both forms of role stress. This study indicates that stress levels are not overwhelming to the NE and may be encouraging development of required skills. Healthcare organizations will continue to evolve and increase in complexity putting pressure on NEs in their many roles. Although this study did not consider strategies for coping with stress, this important avenue needs to be considered as NEs move forward with more extensive roles in the leadership of our healthcare system. PMID- 20023566 TI - The importance of engaging with TERCAP: taxonomy of error root cause analysis and practice-responsibility. PMID- 20023567 TI - Ixabepilone, a new treatment option for metastatic breast cancer. AB - Patients with metastatic breast cancer (MBC) who develop resistance to anthracyclines and taxanes have limited therapeutic options. There is an unmet need for agents that can overcome tumor resistance and are effective in treating drug-resistant disease. The novel epothilone B analog, ixabepilone, binds to beta tubulin and stabilizes microtubules, leading to growth inhibition and apoptosis. Ixabepilone is able to overcome common mechanisms of resistance that limit the efficacy of many chemotherapeutic agents such as anthracyclines, taxanes, and capecitabine. Single-agent ixabepilone has clinical activity and a manageable safety profile against several different solid tumors including MBC, nonsmall cell lung cancer (NSCLC), and prostate cancer. Several phase II and III trials have demonstrated that ixabepilone-as monotherapy and in combination with capecitabine-is active in patients with pretreated or resistant MBC. Ixabepilone is the first epothilone approved for use as monotherapy or in combination with capecitabine in the treatment of metastatic or locally advanced breast cancer that is resistant or refractory to anthracyclines, taxanes, and/or capecitabine. Further clinical studies are evaluating the combination of ixabepilone with targeted agents such as trastuzumab or bevacizumab to further define the role of this novel agent in the treatment of MBC. PMID- 20023568 TI - Prolonged chemoradiation in locally advanced carcinoma of the uterine cervix: final results of a phase II study (ESTER-1). AB - INTRODUCTION: The aim of this phase II study was to evaluate response and toxicity of a prolonged chemoradiation regimen in patients with locally advanced cervical cancer. PATIENTS AND METHODS: Three cycles of concomitant chemotherapy were used with cisplatin (20 mg/m2, 2-hour intravenous infusion, days 1-4) and 5 fluorouracil (1000 mg/m2, 24-hour continuous intravenous infusion, days 1-4) administered at weeks 1, 5, and 9 of radiotherapy. In combination, radiotherapy was delivered to a planning target volume (PTV) defined as the CTV (clinical target volume) plus 8 mm. The CTV was defined as follows: gross tumor volume, upper half of the vagina (if not involved) or the whole vagina (if clinically involved), uterus, obturator nodes, external iliac nodes, internal iliac nodes, and the presacral nodes (cranial to S2). The prescribed dose to the PTV was 50 Gy, 2 Gy/fraction (ICRU 62) delivered in 25 fractions with a 2-week break at 20Gy and 40 Gy (split-course technique). Early and late toxicity was assessed according to the RTOG and RTOG/EORTC toxicity scales. Perioperative toxicity was evaluated according to the Chassagne classification of surgical complications. RESULTS: A total of 25 patients were included in this study. Median age was 52 years (range, 28-69). Clinical stage was: IB2-II: 19 patients (76%), III-IVA: 6 patients (24%). All patients completed the prescribed dose of chemoradiation and were evaluated 4 weeks after the end of treatment. Complete and partial clinical local response was observed in 4 and 19 patients, respectively (totaling 92% of clinical responses). About 32% of patients experienced grade 3 to 4 toxicity, in particular, grade 3 or 4 hematological toxicity was observed in 7 patients and 1 patient developed grade 3 genitourinary toxicity. No patients developed grade 3 gastrointestinal toxicity or skin toxicity. Of total, 22 patients (88%) underwent radical hysterectomy. Seven patients (28%) showed a complete response (CR) to treatment, and 7 patients (28%) showed microscopic residual disease (MUPR), totaling 14 patients (56%) complete/partial microscopic responses. Perioperative morbidity was higher than reported in historical controls especially in terms of tissue fibrosis (64%) and perioperative urinary toxicity (14%). Actuarial 2-year local control, disease-free survival, and overall survival were 65.5%, 61.8%, and 80.8%, respectively. CONCLUSION: A prolonged treatment with more chemotherapy courses does not improve tumor response and increases the risk of perioperative complication. This treatment regimen, considering the low incidence of acute gastrointestinal toxicity, might be tested in the adjuvant setting. PMID- 20023569 TI - Ductal carcinoma in situ: size and resection volume predict margin status. AB - OBJECTIVES: There is strong evidence that breast conservation surgery (BCS) with negative margins for ductal carcinoma in situ (DCIS) is associated with low rates of recurrence. Our goal was to identify factors associated with positive margins in BCS for DCIS. METHODS: A retrospective database review identified 823 patients diagnosed with DCIS. The current analysis included 546 of those patients treated with BCS from 2000 to 2006 with complete data regarding tumor and lumpectomy dimensions. Variables analyzed included tumor size, lumpectomy volume, estrogen and progesterone receptor status, histologic subtype, grade, and age at diagnosis. chi analysis and t tests were used to identify factors that may predict positive margins. A multivariate regression model was developed to determine independent variables predictive of positive margin status. RESULTS: A total of 33% of specimens had positive margins. Lumpectomy volume, tumor size, nuclear grade (low vs. high), and number of slides positive for DCIS were all significant for positive margin status by bivariate analysis. On multivariate analysis, tumor size (P < 0.001; odds ratio, 2.37; 95% confidence interval, 1.712, 3.296) and resection volume (P = 0.0006; odds ratio, 0.48; 95% confidence interval, 0.318, 0.729) remained significantly associated with positive margin status. Age at diagnosis, histologic subtype, tumor grade, and estrogen and progesterone status all were not associated with margin status. CONCLUSIONS: Positive margins after BCS for DCIS are associated with larger lesions and a smaller volume of resection. With 33% of patients having positive margins, these data suggest that a more aggressive initial resection may avoid positive margins and thus lower the risk of recurrence or the need for additional surgery. PMID- 20023570 TI - Relationship between chemotherapy use and cognitive impairments in older women with breast cancer: findings from a large population-based cohort. AB - BACKGROUND: Several small scale clinical trials indicated a possible relationship between chemotherapy administration and the increased risk of cognitive impairments in patients with breast cancer, but little information was available from large population-based cohort studies. METHODS: We studied 62,565 women who were diagnosed with stages I-IV breast cancer at age >=65 years from 1991 through 2002 from 16 regions in the Surveillance, Epidemiology and End Results program who were free of cognitive impairments at diagnosis with up to 16 years of follow up, and also studied 9752 matched cohort based on the propensity of receiving chemotherapy. The cumulative incidence of cognitive impairments was calculated and the time to event (cognitive impairments) analysis was conducted using Cox hazard regression model. RESULTS: Overall, patients who received chemotherapy were 8% more likely to have drug-induced dementia compared with those without chemotherapy, but that was not statistically significant after adjusting for patient and tumor characteristics (hazard ratio = 1.08, 95% confidence interval = 0.85-1.37). The risk of developing Alzheimer disease, vascular dementia, or other dementias was significantly lower in patients receiving chemotherapy except for cognitive disorder which was not significantly different between the 2 chemotherapy groups. The results were somewhat similar in the entire cohort and the matched cohort based on the probability of receiving chemotherapy. CONCLUSION: There was no significant association between chemotherapy and the risk of developing drug-induced dementia and unspecified cognitive disorders. The risk of developing Alzheimer disease, vascular dementia, or other dementias was significantly lower in patients receiving chemotherapy. This study with long-term follow-up did not support the findings that chemotherapy was associated with an increased risk of late stage cognitive impairments. PMID- 20023571 TI - Triple-negative breast cancer: a short review. AB - This review describes the pathology, prognosis, current treatment options, and future directions for the management of patients with triple-negative breast cancer. "Triple-negative" tumors lack expression of estrogen and progesterone receptors, and HER2. The subtype comprises some 15% of all breast cancers, with tumors of a typically larger size and higher grade. Clinically, triple-negative breast cancer has a relatively high rate of recurrence and distant metastasis, and poor overall survival. The standard of care is chemotherapy, although recent research suggests a sound rationale for the use of targeted agents with antitumor and/or antiangiogenic activity such as receptor tyrosine kinase inhibitors. PMID- 20023572 TI - Differential effects of 5-fluorouracil on glucose transport and expressions of glucose transporter proteins in gastric cancer cells. AB - Although 5-fluorouracil (5-FU) is a widely used chemotherapeutic agent in the treatment of gastric cancer, the underlying mechanism for 5-FU resistant phenotype, has yet to be elucidated. We hypothesized that the sensitivity of gastric cancer to 5-FU treatment might be related to the rate of glucose transport (GLUT), and investigated the expressions of GLUT1, 2, 3, and 4 in two different gastric cancer cells (SNU-216, moderately differentiated gastric adenocarcinoma; and SNU-668, signet ring cell gastric carcinoma). Immunohistochemistry of GLUT1 and GLUT4 and immunoblot analysis of glycogen synthase kinase 3 were also performed. Hexokinase activity was measured. We found that 5-FU suppressed glucose uptake in SNU-216, while it stimulated GLUT in SNU 668. Further analysis revealed that 5-FU decreased the expression levels of GLUT1, 2, and 4 in SNU-216 cells and increased the expression levels of GLUT1, 2, and 4 in SNU-668 cells. Consistent with GLUT expression levels, immunohistochemistry analysis showed that 5-FU increased GLUT1 and GLUT4 levels in SNU-216 and decreased GLUT1 and GLUT4 levels in SNU-668. We also observed that glycogen synthase kinase 3 activity was decreased in SNU-216 and increased in SNU 668 with 5-FU treatment. No significant difference in hexokinase activities was observed with 5-FU treatment. Taken together, these results suggest that 5-FU exerts differential effects on GLUT depending on gastric cancer cell types, which may indicate a possible explanation, at least in part, for the differing responses to 5-FU chemotherapy in gastric cancer. PMID- 20023573 TI - Current use of clozapine in Parkinson disease and related disorders. AB - OBJECTIVE: Clozapine has been shown to be effective in the treatment of Parkinson disease psychosis and tremor; however, use of clozapine has been limited because of potential adverse effects and stringent monitoring criteria. METHODS: To evaluate safety and efficacy in a naturalistic setting, we conducted a retrospective chart review of all patients ever treated with clozapine at a movement disorders clinic in Rhode Island. RESULTS: Clozapine was indicated for psychosis in 39 patients, for tremor in 19, and for both tremor and psychosis in an additional 6 patients. Fifty of 61 patients reported improvement in their symptoms. Clozapine use was interrupted in 26 patients, for various reasons including transition to hospice care (3), worsening of motor symptoms (4), agranulocytosis (1), intolerable adverse effects (9), symptom resolution (2), and other or unknown reasons (7). CONCLUSIONS: Clozapine may be useful in the management of movement disorders and related symptoms so long as adequate supervision and monitoring can be provided. PMID- 20023575 TI - Trim long eyebrow hairs in chronic keratitis. PMID- 20023574 TI - A modified deep anterior lamellar keratoplasty using intacs dissectors. AB - PURPOSE: To describe a new technique for deep anterior lamellar keratoplasty in the human eye. METHODS: We obtained globes from the San Diego Eye Bank and performed central and peripheral pachymetry measurements at the 8-mm optical zone. We made a radial incision at the edge of the optical zone at 90% depth of central corneal thickness. We enlarged the incision and started lamellar dissection. The curved tunnel was created using dissecting instruments (Addition Technology, Des Plaines, IL), used in Intacs channels. With a sweeping motion, we connected these channels. An 8-mm Barron Radial Vacuum Trephine (Baron Precision Instruments, LLC, Grand Blanc, MI) was placed over the optical zone until the 90% depth of thickness was reached. The cap tissue was detached and viscoelastic was used to separate Descemet membrane and stromal tissue. We photographed the surface for analysis. RESULTS: This technique was easy to perform with little remaining posterior stromal tissue. Residual tissue could easily be dissected, and the vacuum trephine could easily reach the same depth of the pocket. The outer edge of the bed had a clean sharp edge. The stromal bed surface and the inner corneal cap were smooth. CONCLUSIONS: We conclude that this modified technique is easy to perform and results in a more regular stromal surface. This technique may reduce the need for penetrating keratoplasty. PMID- 20023576 TI - Transfer of a T-helper type 2 clone into conjunctiva induces corneal damage in mice. AB - PURPOSE: We established a T-helper Type 2 (Th2) clone-induced conjunctival eosinophilia model by injecting D10.G.4.1 (D10) cells, a murine Th2 clone, and conalbumin, its specific antigen, into conjunctiva of AKR/J mice. Using this model, we investigated the effect of a coinjection of D10 cells and conalbumin into conjunctiva on corneal damage. METHODS: Corneal fluorescein staining scores and eosinophil peroxidase (EPO) activity in conjunctiva were measured after coinjection of D10 and conalbumin into conjunctiva, and the effects of cyclosporine A, betamethasone, and anti-interleukin-5 antibody on staining scores and EPO activity were examined. RESULTS: Coinjection of D10 and conalbumin induced an increase of the corneal fluorescein staining score after 24, 48, and 96 hours and 10 days. EPO activity in conjunctiva increased time-dependently until 24 hours after coinjection. The increase in the staining score followed the time dependent increase in EPO activity. The instillation of cyclosporine A, an inhibitor of cytokine production from T-cells, and betamethasone significantly inhibited the increase in corneal fluorescein score and EPO activity. Intraperitoneal administration of anti-interleukin-5 monoclonal antibody, which inhibits the infiltration of eosinophils into the conjunctiva, completely inhibited the increase in staining score. CONCLUSION: The transfer of the Th2 clone into the murine conjunctiva induced corneal damage, which may have been caused by Th2 cell-produced interleukin-5 that mediated the activation of eosinophils. PMID- 20023577 TI - Comparative analysis of carmellose 0.5% versus hyaluronate 0.15% in dry eye: a flow cytometric study. AB - PURPOSE: To compare the effects of Viscofresh 0.5% (carmellose sodium 0. 5%) versus Lubristil (sodium hyaluronate 0.15%) in dry eye syndrome and to study the influence of these two treatments on the expression of various inflammatory markers by flow cytometry in impression cytology specimens. METHODS: In this randomized, masked-observer, parallel group, single-center study, 15 patients with dry eye syndrome were randomized to sodium carmellose 0.5% or sodium hyaluronate 0.15% 1-month treatment after a 1-week washout period. Corneal staining with flurescein, breakup time, Schirmer 1 test with anesthesia (Jones test), and tear clearance were assessed. Besides, conjunctival impression cytology was performed to investigate inflammatory markers (CD3, CD11b, and HLA DR) using flow cytometry. RESULTS: Carmellose group shows statistical improvement compared with the hyaluronate group in breakup time, corneal staining, and HLA DR. The two other inflammatory markers had also a tendency for a decreased expression in both groups, with no statistical significance. There were neither visual acuity loss nor other complications related to treatment. CONCLUSION: Both artificial tears improve dry eye signs and symptoms and inflammatory markers expression, with significant better results in carmellose group. PMID- 20023578 TI - Central corneal thickness in children with intellectual disability: a controlled study. AB - PURPOSE: To evaluate the central corneal thickness (CCT) in children with intellectual disability (ID) and to determine the association between ID-related variables and CCT. METHODS: A total of 77 subjects with ID and 38 healthy controls were included in the study. The subjects with ID were subdivided into mild (IQ 50-69; n = 34), moderate (IQ 35-49; n = 30), and severe (IQ <34; n = 13) groups and syndromic (n = 40) versus nonsyndromic (n = 37) distinctions. All children underwent CCT assessment by ultrasound pachymetry, in addition to complete ophthalmologic examination. Analysis of variance, chi test, and linear regression analysis were used for statistical analysis. RESULTS: CCT was significantly higher in the ID group (549.7 +/- 21.4 microm) than that the control group (521.6 +/- 16.9 microm; P < 0.0001). Linear regression analysis revealed that presence of syndromic etiology significantly predicted higher CCT among the individuals with ID (P < 0.0001). CONCLUSIONS: Children with ID have an increased CCT compared with healthy controls. Syndromic etiology is the only variable that predicts higher CCT among the individuals with ID. Although the finding of an increased CCT in children with ID is statistically significant, it is not clinically significant. PMID- 20023579 TI - Deep anterior lamellar keratoplasty in patients with keratoconus: big-bubble technique. AB - PURPOSE: To evaluate the visual and refractive outcomes after deep anterior lamellar keratoplasty (DALK) using the big-bubble technique in eyes with keratoconus. METHODS: In this case series, eyes with moderate to advanced keratoconus underwent DALK. All of them had unacceptable spectacle-corrected visual acuity or were contact lens intolerant. DALK was performed using the big bubble technique. Full thickness donor cornea without Descemet's membrane (DM) was sutured to the recipient bed with three different suturing techniques. The visual acuity (presented in logMAR), refractive status, intraoperative and postoperative complications were evaluated. RESULTS: A total of 129 consecutive eyes of 121 patients were included. In 3 eyes, the operation was converted to penetrating keratoplasty. Therefore, 126 eyes (79 male) were enrolled in the study. Mean patient age at the time of surgery was 26.08 +/- 7.6 years. Mean follow-up period was 21.62 +/- 9.0 months. Mean preoperative corrected visual acuity was 1.27 +/- 0.4 logMAR, increasing to 0.25 +/- 0.2 logMAR at final follow up examination (P < 0.001). Postoperative mean spherical equivalent refractive error, refractive and keratometric astigmatism were -3.41 +/- 3.1 D, 3.04 +/- 2.3 D, and 3.67 +/- 2.1 D, respectively. Bared DM was achieved in 103 eyes, whereas in 23 eyes pre-Descemet dissection was performed. DM perforation occurred in 5 eyes. The main complications encountered were filamentary keratitis, subepithelial graft rejection, and astigmatism. CONCLUSIONS: : DALK using the big bubble technique appears to be a safe and effective procedure for eyes with moderate to advanced keratoconus. In case of extensive intraoperative DM perforation, it does not pose any limitation to ongoing penetrating keratoplasty. PMID- 20023580 TI - Skepticism and science. PMID- 20023582 TI - Conjunctival melanoma arising from primary acquired melanosis in a patient with neurofibromatosis type I. AB - PURPOSE: To describe a case of conjunctival malignant melanoma (MM) arising from primary acquired melanosis in a patient with neurofibromatosis type 1 (NF-1). METHODS: Case report and literature review. RESULTS: A 66-year-old woman with a history of NF-1 presented with extensive pigmentation of the left bulbar conjunctiva. Conjunctival biopsies demonstrated MM arising from primary acquired melanosis with atypia. Two excision and cryotherapy procedures did not completely eradicate the conjunctival pigment, which was then treated with topical mitomycin C. Subsequent biopsies and clinical examinations have revealed no remaining tumor. CONCLUSION: Conjunctival MM is uncommon in patients with NF-1 and can be successfully treated with excision, cryotherapy, and topical mitomycin C. PMID- 20023583 TI - Conductive keratoplasty followed by collagen cross-linking with riboflavin-UV-A in patients with keratoconus. AB - PURPOSE: To evaluate the combined effect of conductive keratoplasty (CK) followed by corneal collagen cross-linking (CXL) in 2 patients with keratoconus. METHODS: CK spots were applied on the flatter side of the cornea followed by CXL using riboflavin and UV-A light. RESULTS: Immediately after CK, a significant corneal topographic improvement was observed. The CK effect regressed 3 months postoperatively and remained unchanged until the sixth postoperative month in both patients. CONCLUSION: Corneal remodeling with CK in patients with keratoconus seems to have a temporary effect despite the subsequent application of CXL. PMID- 20023584 TI - New INTACS SK implantation in patients with post-laser in situ keratomileusis corneal ectasia. AB - In this case series, 2 patients with post-laser in situ keratomileusis corneal ectasia were included. Patients were treated with new intrastromal corneal ring segments, INTACS SK (severe keratoconus or steep "K") (Addition Technology, Inc, Des Plaines, IL) implantation. Two segments were inserted without any intraoperative or postoperative complications. Three months after the procedure, uncorrected visual acuity was improved in both patients from counting fingers preoperatively to 20/40 and 20/50, respectively. Best spectacle-corrected visual acuity improved from 20/40 to 20/32 and from 20/50 to 20/40. Manifest refraction improved from -8.75/-5.00 x 125 and -14.25/-6.50 x 33 preoperatively to -5.50/ 2.25 x 125 and -8.25/-1.50 x 15 with an improvement in topographic findings. One year postoperatively, uncorrected visual acuity, best spectacle-corrected visual acuity, and topographic findings remained improved. In conclusion, new INTACS SK might be able to improve visual acuity in patients with post-laser in situ keratomileusis corneal ectasia. PMID- 20023585 TI - Concurrent acanthamoeba and Fusarium keratitis with silicone hydrogel contact lens use. AB - PURPOSE: To report a case of simultaneous Acanthamoeba and Fusarium keratitis associated with no-rub multipurpose contact lens solution and silicone hydrogel contact lens use. METHOD: Observational case report. RESULTS: A 39-year-old woman was referred for worsening of a presumed bacterial corneal ulcer in the setting of silicone hydrogellens wear with occasional overnight wear, no-rub multipurpose contact lens solution use, and combined topical antibiotic/corticosteroid treatment. Initial corneal scrapings and culture confirmed Acanthamoeba and Fusarium solani, corroborated by in vivo confocal microscopy findings, yet despite topical chlorhexidine 0.02%, propamidine 1%,neomycin/polymyxin B ointment, and natamycin 5% along with oral itraconazole, the ulcer worsened. Four days after amoebic and fungal therapy initiation, it was discovered that the pharmacy accidentally dispensed neomycin/polymyxin B/dexamethasone, and despite immediate discontinuation, therapeutic penetrating keratoplasty from corneal melting was ultimately required. Corneal histopathology confirmed the presence of amoebic cysts and fungal elements. CONCLUSIONS: Coexisting infection with Acanthamoeba and Fusarium species can occur in contact lens wear. Atypical infection must be considered in patients with corneal ulcers demonstrating poor therapeutic response in the setting of contact lens wear. Corticosteroids should be used with extreme caution in contact lens-related corneal infections, especially when the diagnosis remains unknown because they can lead to acceleration of active infection and keratolysis. PMID- 20023586 TI - Absence of pathogenic mutations in VSX1 and SOD1 genes in patients with keratoconus. AB - PURPOSE: Keratoconus (KC) is a bilateral, noninflammatory, and progressive corneal ectasia that occurs mostly as a sporadic disorder, but it has long been recognized that a significant minority of patients also exhibit a family history. In recent years, several candidate genes, including VSX1 and SOD1, have been proposed and some disease-causing mutations have been identified. METHODS: To investigate the role of the 2 genes in 113 Slovenian patients with sporadic and familial KC, the complete coding region with corresponding intronic sequences was analyzed. The same regions of both genes were also checked in 100 healthy blood donors. We also checked the relation of 627+23G>A polymorphism in the VSX1 gene with the hereditary form of the disease. RESULTS: No disease-causing mutations were identified in either gene. We did discover a significant association of 627+23G>A polymorphism distribution (VSX1) with unrelated patients diagnosed with the hereditary form of KC. CONCLUSION: The absence of pathogenic mutations in our large number of unrelated patients with KC indicates that other genetic factors are involved in the development of this disorder. PMID- 20023587 TI - Mucous membrane grafting for lid margin keratinization in Stevens-Johnson syndrome: results. AB - PURPOSE: To study the efficacy of mucous membrane grafting asa surgical technique to address lid margin keratinization, an important cause for chronic blink related microtrauma in Stevens-Johnson syndrome (SJS). METHODS: The keratinized strip of conjunctiva along the lid margin was excised and replaced by lip mucous membrane using fibrin glue in 54 eyes of 31 patients with lid margin keratinization after SJS. This was a retrospective case series carried out between April 2005 and November 2006. RESULTS: In 50 of 54 eyes (92.6%), there was improvement or stabilization in patient comfort, conjunctival hyperemia, ocular surface staining characteristics, and best-corrected visual acuity over a mean follow-up period of 6 months. CONCLUSIONS: The authors describe the method of mucous membrane grafting for addressing lid margin keratinization, one of the important causes for persistent inflammation in SJS. This procedure not only improves patient comfort and visual acuity by reducing surface inflammation but also possibly prevents further deterioration of ocular surface in SJS. PMID- 20023588 TI - Beauveria keratitis and biopesticides: case histories and a random amplification of polymorphic DNA comparison. AB - PURPOSE: The purposes of this study were to describe 2 contact lens-associated Beauveria keratitis cases and to compare the isolates of 3 contact lens associated Beauveria keratitis cases with Beauveria-based biopesticides using random amplification of polymorphic DNA (RAPD). METHODS: A 55-year-old diabetic woman from New Mexico and a 31-year-old healthy woman from southern Wisconsin developed soft contact lens-related corneal ulcers unresponsive to topical moxifloxacin and prednisolone acetate drops. Their corneal cultures grew B. bassiana. These isolates, an isolate from a third soft contact lens-related Beauveria keratitis case, and Beauveria-based biopesticides sold in the United States were analyzed using morphological features, DNA sequencing, and RAPD. A PubMed, Cochrane Library, OVID, UpToDate, and Google search using the term "Beauveria" found only 9 reported Beauveria keratitis infections. RESULTS: Patient 1 responded to topical natamycin, ketoconazole, and 200 mg oral ketoconazole twice daily before developing a secondary bacterial infection requiring penetrating keratoplasty. After subsequent cataract surgery, the best corrected visual acuity was 20/20. Patient 2 was treated with topical natamycin, topical amphotericin, and 200 mg oral voriconazole twice daily for 1 month with residual scarring and a best-corrected visual acuity of 20/25. RAPD showed that all isolates were unrelated. CONCLUSIONS: Although earlier reported Beauveria keratitis cases occurred after corneal injury in patients who did not wear contact lenses, 3 recent patients wore soft contact lenses and denied trauma, mirroring a changing trend in microbial keratitis. RAPD analysis showed that the Beauveria isolates were unrelated to one another and to Beauveria-based biopesticides. In Patient 2, oral voriconazole worked well. PMID- 20023589 TI - Endothelial keratoplasty: endothelial cell loss after deep lamellar endothelial keratoplasty with retention of an open-loop anterior chamber intraocular lens. AB - PURPOSE: To prospectively assess the impact of a retained open-loop anterior chamber intraocular lens (ACIOLs) on endothelial cell loss after deep lamellar endothelial keratoplasty (DLEK). METHODS: Prospectively gathered central endothelial cell densities of eyes with open-loop ACIOLs after DLEK were examined at 6 months, 1 year, and 2 years. RESULTS: Nine eyes with an open-loop ACIOL were examined. Mean endothelial cell loss was 37% at 6 months (n = 9, P = 0.001), 36% at 1 year (n = 6, P = 0.001), and 41%, at 2 years (n = 5, P = 0.002) after surgery. CONCLUSION: Postoperative endothelial cell loss over the first 2 years in this small series of patients undergoing DLEK with retention of an open-loop ACIOL may be greater in the first 6-12 months but similar at 2 years when compared with that found in the literature for the DLEK procedure. PMID- 20023590 TI - UV-A Collagen Cross-Linking Treatment of Bullous Keratopathy Combined With Corneal Ulcer. AB - PURPOSE: To evaluate the therapeutic effect of corneal cross-linking (CXL) in 2 cases of bullous keratopathy combined with corneal ulcer. METHODS: Two patients (2 eyes) were recruited for the sake of the study. Both suffered from bullous keratopathy and presented a gradually deteriorating, vision-threatening, central corneal ulcer, despite intense local antibiotic therapy. The same surgical procedure was performed in both eyes. De-epithelialization of the affected corneas was accompanied by UV-A cross-linking and finally by the application of a therapeutic contact lens. Local antibiotic therapy was resumed after the procedure. RESULTS: Within 24 hours of the treatment, both patients reported significant subjective improvement of their visual acuity and ocular discomfort. Clinical evaluation revealed improvement of the corneal ulcer and the bullous keratopathy associated with significant decrease of the corneal thickness and haziness. During the 2-month follow-up period, a significant improvement of visual acuity was recorded in both cases. CONCLUSION: CXL should be considered as a potential adjuvant therapeutic tool in patients with combined bullous keratopathy and infectious keratitis, who are resistant to traditional topical therapy. PMID- 20023591 TI - Nondisplacement of descemet stripping automated endothelial keratoplasty lenticule after traumatic rupture of the globe. AB - PURPOSE: The purpose of this study was to report a case of nondisplacement of an endothelial keratoplasty (EK) lenticule after traumatic rupture of the globe. METHODS: The authors conducted a review of the clinical course of a 35-year-old woman who sustained a traumatic rupture of the globe 4 months after Descemet stripping endothelial keratoplasty. RESULTS: The rupture did not involve the 4 month-old temporal scleral tunnel wound used for the EK. The lenticule was attached despite the injury and remained so in the postoperative course. CONCLUSIONS: Blunt trauma after endothelial keratoplasty can result in the disruption of the operative wound and lenticule dislocation. However, in our case, the rupture site did not involve the surgical wound, and the lenticule remained attached throughout the trauma, the surgical repair of the ruptured globe, and the postinjury course. This case illustrates the potential strength of EK wounds and the strength of adhesion of the transplanted lenticule and poses unanswered questions about the process of wound healing and attachment of donor lenticules in EK cases. PMID- 20023592 TI - Small animal tumour imaging with MRI and the ECAT EXACT scanner: application of partial volume correction and comparison with microPET data. AB - OBJECTIVE: Partial volume effects caused by limited spatial resolution of conventional positron emission tomography (PET) scanners result in an underestimation of the activity concentration in small tumours. The aim of the study was to evaluate the feasibility of small animal tumour imaging with the clinical PET scanner ECAT EXACT after partial volume correction based on MRI calculations. The same tumour model was examined additionally with the small animal PET system, microPET focus 120. METHODS: Before the ECAT EXACT studies recovery coefficients for different sphere volumes were generated with phantom experiments. For the following in-vivo study DS-sarcoma cells were implanted on both hind foot dorsum of male Sprague-Dawley rats. In-vivo tumour volume calculations were done with the high-resolution MRI system, Magnetom Vision Experimental. Dynamic F-fluorodeoxyglucose (FDG) PET was performed with the scanner ECAT EXACT (5 MBq intravenous, two-dimensional mode, n = 16 tumours) or with the microPET focus 120 (20 MBq intravenous, two-dimensional mode, n = 10 tumours). The animals were then killed, the tumours rapidly explanted, weighed and homogenized. The concentration of F-FDG was measured with a gamma counter and decay corrected; the ex-vivo F-FDG concentration was compared with the mean tumour activity concentration of the PET data. RESULTS: Using the ECAT EXACT mean underestimation of actual tumour F-FDG concentration was 35.4%, for partial volume-corrected data this error decreased to 1.7%. In addition, after partial volume correction congruence and linear correlation between the regions of interest-based activity concentration and ex-vivo measurements were excellent (r = 0.98). These results were quite similar to the microPET experiments without partial volume correction: r = 0.99. CONCLUSION: These data indicate that partial volume correction might allow use of the clinical PET system, ECAT EXACT, for the metabolic assessment of small animal tumours >/=10 mm with sufficient accuracy if no dedicated animal PET is available. PMID- 20023593 TI - The effect of acquisition time on visual and semi-quantitative analysis of F-18 FDG-PET studies in patients with head and neck cancer. AB - OBJECTIVE: To evaluate the impact of acquisition time on F-18 fluorodeoxyglucose positron emission tomography (PET) image quality, lesion detection rate, standard uptake and lesion volume measures. METHODS: In 17 consecutively referred head and neck cancer patients, a 10-min acquisition of the head and neck was performed in list mode after completion of a whole-body PET-computed tomography scan. For each patient, 0.5, 1, 2, 3, 5, 7 and 10-min PET images were reconstructed. Image quality was scored on a 5-point scale. Lesions were visually identified and lesion locations were compared between scans with various acquisition times. Standard uptake values (SUVs) and lesion volumes were automatically obtained and compared. RESULTS: Image quality was scored excellent/good for 10 and 7-min scans; moderate for 5 and 3-min scans and poor/very poor for 2, 1 and 0.5-min scans. Lesion detection rate did not differ between scans with shorter acquisition times and the reference. Similarly, SUVmax was not significantly different between images with various scan times and the reference, except for the 0.5-min PET image (Wilcoxon, P = 0.04). Overall, lesion volume increased significantly with longer scan time (Friedman, P<0.0001). For individual lesions, however, the effect of scan time on volume was quite variable. The variability was significantly larger for images with scan times below 3 min than for images with scan times of > or = 3 min. CONCLUSION: Despite poor image quality for scans with short acquisition times, the lesion detection rate was not adversely affected. Increased scan time resulted in an increase in lesion volumes. This finding is of particular interest if PET images are implemented in radiation oncology treatment planning. PMID- 20023595 TI - Gene variations in the cholecystokinin system in patients with panic disorder. AB - OBJECTIVES: Panic disorder (PD) is a common psychiatric disease occurring more frequently in women than men. Multiple common and/or rare variants in the genome contribute to the complex etiology of the disorder. The neuropeptide cholecystokinin (CCK) and its receptors (the CCK system) have been suggested to be involved in the pathogenesis of PD. METHODS: We examined the promoter, exon, and exon-intron boundaries of the genes encoding CCK and its receptors (CCKAR and CCKBR) for variations in 187 patients with PD and 277 screened control individuals. Up to 1342 additional healthy population controls were examined for some of the variations. One CCK gene intron variation was analyzed for alternative splicing using an exon-trapping assay. RESULTS: The promoter variant (-36C > T; rs1799923) and an intron 1 polymorphism (IVS1-7C > G; rs754635) in the CCK gene were found to protect against PD (P<0.05). The intron 1 variation did not seem to alter the splicing of the gene. None of the other variations found were associated with PD, but a 2-marker haplotype (rs1800855/rs1800857) in the CCKAR gene protected women against PD (P=0.004). In addition, we found two novel rare missense variations in the CCKBR gene (Lys329Asn and Pro446Leu) in two and one patient, respectively. CONCLUSION: The results suggest that the CCK system may play a role in the pathogenesis of PD, with susceptibility alleles both protecting and contributing to the disease. Both common and rare variants seem to be involved. The involvement of the CCK system may also contribute to the increased prevalence of PD in women. PMID- 20023594 TI - Bisphosphonates pathway. PMID- 20023596 TI - Identifying unreported and undiagnosed cases of congenital syphilis in Arizona using live birth and fetal death registries. AB - To investigate the drop in reported congenital syphilis cases from 28 in 2005 to 16 in 2006, the Arizona infant registries were cross matched with reported syphilis test among women in th estat STD database. Six previously unreported cases were identified; four live births and two still births. PMID- 20023597 TI - Gastric syphilis: a systematic review of published cases of the last 50 years. AB - The authors conducted a systematic review of the English literature for cases of Gastric Syphilis (GS) in the last 50 years. The 34 studies which met selection criteria included 52 patients with GS. Of the reviewed patients, only 13% had a history of syphilis diagnosis and 46% had prior or concurrent clinical manifestations of the disease. Epigastric pain/fullness was the most common presenting symptom (92%) and epigastric tenderness being the most common sign. Gastric bleeding of variable intensity was documented in 35% of the cases. In the radiologic examinations, fibrotic narrowing and rigidity of the gastric wall was the most common finding (43%), followed by hypertrophic and irregular folds, while in endoscopy the most common lesion types were multiple ulcerations (48%), nodular mucosa, and erosions. The antrum was the most commonly affected area (56%). The majority of the patients received penicillin (83%) with a rapid resolution of their symptoms. Seventeen percent of the patients were treated surgically either due to a complication or due to strong suspicion of infiltrating tumor or lymphoma. The nonspecific clinical, radiologic, and pathologic characteristics of GS can establish it as a great imitator of other gastric diseases. GS should be considered in the differential diagnosis in patients at risk for sexually transmitted diseases who present with abdominal complaints and unusual endoscopic lesions and no other diagnosis is made, irrespective of the presence of H. pylori. The absence of primary or secondary luetic lesions should not deter one from considering GS. PMID- 20023598 TI - Prevalence and correlates of Trichomonas vaginalis among incarcerated persons assessed using a highly sensitive molecular assay. AB - We describe the epidemiology of Trichomonas vaginalis (TV) among San Francisco County Jail inmates using APTIMA TV analyte-specific reagents on remnant urine. We detected TV in 15/713 (2.1%) men and 95/297 (32.0%) women. Among women, increased age was significantly associated with TV. The benefits of TV screening should be determined. PMID- 20023599 TI - Systemic administration of a centrally acting acetylcholinesterase inhibitor improves outcome from hemorrhagic shock during acute alcohol intoxication. AB - Previously, we have demonstrated that acute alcohol intoxication impairs hemodynamic counter-regulation to hemorrhage in unanesthetized rats, and that this phenomenon is associated with an impaired neuroendocrine response to blood loss. Moreover, we demonstrated that central acetylcholinesterase inhibition restores the hemodynamic and neuroendocrine responses to hemorrhage in alcohol intoxicated rats. We hypothesized that similar responses could be elicited by systemic administration of physostigmine, an acetylcholinesterase inhibitor that penetrates the blood brain barrier. The relevance of this approach was to establish effectiveness of a more clinically applicable route of drug administration than that used previously. Chronically catheterized adult male Sprague-Dawley rats (250-275 g) were administered a bolus of physostigmine (i.v., 100 microg/kg) at rest, and in a separate study, simultaneously with Ringer's lactate solution after an overnight intragastric infusion of 30% alcohol (approximately 7 g/kg for 15 h) or 52% isocaloric dextrose and fixed-pressure hemorrhage. I.v. physostigmine administration immediately increased sympathetic outflow via activation of central nicotinic receptors and improved the pressor response to fluid resuscitation in both dextrose controls and alcohol-intoxicated animals. The improved hemodynamic recovery achieved with physostigmine was also associated with attenuation of the rises in the markers of liver and renal damage alanine aminotransferase and blood urea nitrogen in alcohol-intoxicated animals. Additional studies are warranted to determine the effect of central acetylcholinesterase inhibition on tissue injury and survival after severe blood loss, as well as its effects on long-term metabolic and inflammatory responses. PMID- 20023600 TI - The P38alpha and P38delta MAP kinases may be gene therapy targets in the future treatment of severe burns. AB - Microvascular barrier damage, induced by thermal injury, imposes life-threatening problems owing to the pathophysiological consequences of plasma loss and impaired perfusion that finally may lead to multiple organ failure. The aim of the present study was to define the signaling role of selected mitogen-activated protein kinases (MAPKs) in general vessel hyperpermeability caused by burns and to look for a potential gene therapy. Rearrangement of cytoskeletons and cell tight junctions were evaluated by phalloidin labeling of actin and immunocytochemical demonstration of the ZO-1 protein, whereas blood vessel permeability was evaluated by a fluorescence ratio technique. The p38 MAPK inhibitor SB203580 largely blocked burn serum-induced stress-fiber formation and tight-junction damage. Using the adenoviral approach to transfect dominant negative forms of p38 MAPKs, we found that p38alpha and p38delta had similar effects. The in vivo part of the study showed that transfection of these two constructs significantly lowered general venular hyperpermeability and enhanced the survival of burned animals. Because the p38 MAPK pathway seems to play a crucial role in burn induced vascular hyperpermeability, general transfection with p38 MAP dominant negative constructs might become a new therapeutic method to block burn-induced plasma leakage. PMID- 20023601 TI - Deficiency of Bid protein reduces sepsis-induced apoptosis and inflammation, while improving septic survival. AB - Increased apoptotic cell death is believed to play a pathological role in patients with sepsis and experimental animals. Apoptosis can be induced by either a cell death receptor (extrinsic) or a mitochondrial (intrinsic) pathway. Bid, a proapoptotic member of the Bcl-2 family, is thought to mediate the cross talk between the extrinsic and intrinsic pathways of apoptosis; however, little is known about the action of Bid in the development of apoptosis and organ-specific tissue damage/cell death as seen in polymicrobial sepsis. Our results show that after the onset of sepsis, tBid (the active form of Bid) is significantly increased in mitochondrial fractions of the thymus, spleen, Peyer patches, and liver, and that Fas or FasL deficiency blocks Bid activation in various tissues after septic challenge. Increased Bid activation is correlated with increased active caspase-3, caspase-9, and apoptosis during sepsis. Bid-deficient mice exhibit significantly reduced apoptosis in the thymus, spleen, and Peyer patches compared with background mice after sepsis. Furthermore, Bid-deficient mice had significantly reduced systemic and local inflammatory cytokine levels and improved survival after sepsis. These data support not only the contribution of Bid to sepsis-induced apoptosis and the onset of septic morbidity/mortality, but also the existence of a bridge between extrinsic apoptotic signals, e.g., FasL:Fas, TNF:TNFR, and so on, and the intrinsic mitochondrial pathway via Bid tBid activation during sepsis. PMID- 20023602 TI - Notoginsenoside R1 attenuates renal ischemia-reperfusion injury in rats. AB - Ischemia-reperfusion (I/R) injury of the kidney is a complex pathophysiological process and a major cause of acute renal failure. It has been shown that I/R injury is related to inflammatory responses and activation of apoptotic pathways. Inhibition of certain elements of inflammatory responses and apoptotic pathway seemed to ameliorate renal I/R injury. As an effective element of Panax notoginseng, NR1 has antioxidant, anti-inflammatory, antiapoptotic, and immune stimulatory activities. Therefore, we speculate that NR1 can attenuate renal I/R injury. Ischemia-reperfusion injury was induced by renal pedicle ligation followed by reperfusion along with a contralateral nephrectomy. Male Sprague Dawley rats were randomized to four groups: sham group, I/R control group, NR1-1 group (rats treated with NR1, 20 mg.kg.d) and NR1-2 group (rats treated with NR1, 40 mg.kg.d). All animals were killed 72 h after I/R induction. Blood and renal tissues were collected. Renal dysfunction was observed by the level of serum creatinine and histological evaluation. Apoptosis and inflammatory response in the tissue of kidney were detected mainly with molecular biological methods. NR1 attenuated I/R-induced renal dysfunction as indicated by the level of serum creatinine and histological evaluation. It prevented the I/R-induced increases in the levels of proinflammatory cytokine TNF-alpha, myeloperoxidase activity, phosphorylation of p38, and activation of nuclear factor kappaB with cell apoptosis in the kidney and enhanced expression of antiapoptosis cytokine bcl-2. Treatment with NR1 improves renal function after I/R associated with a significant reduction in cell apoptosis and inflammatory responses, which may be related to p38 and nuclear factor kappaB inhibition. PMID- 20023603 TI - The impact of workers' compensation on outcomes of surgical and nonoperative therapy for patients with a lumbar disc herniation: SPORT. AB - STUDY DESIGN: Prospective randomized and observational cohorts. OBJECTIVE: To compare outcomes of patients with and without workers' compensation who had surgical and nonoperative treatment for a lumbar intervertebral disc herniation (IDH). SUMMARY OF BACKGROUND DATA: Few studies have examined the association between worker's compensation and outcomes of surgical and nonoperative treatment. METHODS: Patients with at least 6 weeks of sciatica and a lumbar IDH were enrolled in either a randomized trial or observational cohort at 13 US spine centers. Patients were categorized as workers' compensation or nonworkers' compensation based on baseline disability compensation and work status. Treatment was usual nonoperative care or surgical discectomy. Outcomes included pain, functional impairment, satisfaction and work/disability status at 6 weeks, 3, 6, 12, and 24 months. RESULTS: Combining randomized and observational cohorts, 113 patients with workers' compensation and 811 patients without were followed for 2 years. There were significant improvements in pain, function, and satisfaction with both surgical and nonoperative treatment in both groups. In the nonworkers' compensation group, there was a clinically and statistically significant advantage for surgery at 3 months that remained significant at 2 years. However, in the workers' compensation group, the benefit of surgery diminished with time; at 2 years no significant advantage was seen for surgery in any outcome (treatment difference for SF-36 bodily pain [-5.9; 95% CI: -16.7-4.9] and physical function [5.0; 95% CI: -4.9-15]). Surgical treatment was not associated with better work or disability outcomes in either group. CONCLUSION: Patients with a lumbar IDH improved substantially with both surgical and nonoperative treatment. However, there was no added benefit associated with surgical treatment for patients with workers' compensation at 2 years while those in the nonworkers' compensation group had significantly greater improvement with surgical treatment. PMID- 20023605 TI - Patients' preferences for scoliosis brace treatment: a discrete choice experiment. AB - STUDY DESIGN: Discrete choice experiment. OBJECTIVE: To investigate the reduction in the risk of surgery that scoliosis patients would require in order to consider brace treatment as acceptable, and to elicit the trade-offs individuals make between characteristics of brace treatment. SUMMARY OF BACKGROUND DATA: The effectiveness of brace treatment in idiopathic scoliosis patients has not been established in randomized controlled trials (RCTs). Treatment with a brace can be quite bothersome. Patients' preferences for brace treatment are unknown. Insight into patients' preferences for (characteristics of) brace treatment will be useful for future trials and for the development of braces that may optimize compliance with brace treatment. METHODS: A total of 197 patients who had completed treatment (brace and/or surgery) for scoliosis were approached for the study, of which 135 gave informed consent. A discrete choice experiment was designed in which patients had to choose between hypothetical brace treatment profiles that differed in following 4 treatment attributes: effectiveness, visibility, discomfort, and treatment duration. A multinomial logit model was used to analyze the relative importance of these attributes. Subgroup analyses were conducted for brace-only, brace-surgery, and surgery-only patients. RESULTS: The response rate was 86% (116/135). All treatment attributes proved to be important for patients' choices. All subgroups were prepared to initiate treatment with a Boston brace if the brace would reduce the need for surgery by 53%. Risk reductions in a range of 32% to 74% were required for acceptance of a treatment duration of 3 years. CONCLUSION: Scoliosis patients stated to be prepared to undergo brace treatment only if it provides sizeable reduction of the risk of surgery. Effectiveness and discomfort in wearing a brace were the most important determinants of the choices. These results are important if RCTs would conclusively establish that bracing is effective, and show directions for the further technical development of braces to increase the compliance with brace treatment. PMID- 20023606 TI - Anterior cervical decompression and fusion accelerates adjacent segment degeneration: comparison with asymptomatic volunteers in a ten-year magnetic resonance imaging follow-up study. AB - STUDY DESIGN: Prospective 10-year follow-up magnetic resonance imaging (MRI) study of patients who underwent anterior cervical decompression and fusion (ACDF) and healthy control subjects. OBJECTIVE: To clarify the incidence of adjacent segment degeneration during 10 years after ACDF. SUMMARY OF BACKGROUND DATA: There have been few studies which investigated incidence of progression of degenerative changes at adjacent segments in patients treated by ACDF comparing with healthy subjects. METHODS: Sixty-four patients who underwent ACDF (48 males, 16 females, mean age 47.3 years, mean follow-up 12.1 year) and 201 asymptomatic volunteers who underwent MRI in our previous study (113 males, 88 females, mean age; 41.1 year, mean follow-up; 11.7 years) were included in this study. The patients and control subjects underwent follow-up MRI in this study. Following MR findings were evaluated using a numerical grading system from C2-C3-C7-T1: (1) Decrease in signal intensity of disc (DSI), (2) Posterior disc protrusion (PDP), (3) Disc space narrowing, and (4) Foraminal stenosis. When an increase in at least one grade in any of the radiographic parameters was detected between the 2 time points, progression of disc degeneration was judged as present at the level of interest. RESULTS: Progression of DSI was significantly more frequent in ACDF group than in control group at C4-C5, while progression of PDP was significantly more frequent in ACDF group than in control group at all levels except for C5-C6. Progression of disc space narrowing and foraminal stenosis was significantly more frequent in ACDF group at C3-C4 and at C6-C7, respectively. CONCLUSION: Although both ACDF patients and control subjects demonstrated progression of disc degeneration during 10 years, ACDF patients had significantly higher incidence of progression of disc degeneration at adjacent segments than control subjects, while progression of disc degeneration at adjacent segments was not always related to development of clinical symptoms. PMID- 20023607 TI - The effect of backpacks on the lumbar spine in children: a standing magnetic resonance imaging study. AB - STUDY DESIGN: This study is a repeated measures design to measure the lumbar spine response to typical school backpack loads in healthy children. The lumbar spine in this setting was measured for the first time by an upright magnetic resonance imaging (MRI) scanner. OBJECTIVE: The purpose of this study is to measure the lumbar spine response to typical school backpack loads in healthy children. We hypothesize that backpack loads significantly increase disc compression and lumbar curvature. SUMMARY OF BACKGROUND DATA: Children commonly carry school backpacks of 10% to 22% bodyweight. Despite growing concern among parents about safety, there are no imaging studies which describe the effect of backpack loads on the spine in children. METHODS: Three boys and 5 girls, age 11 +/- 2 years (mean +/- SD) underwent T2 weighted sagittal and coronal MRI scans of the lumbar spine while standing. Scans were repeated with 4, 8, and 12 kg backpack loads, which represented approximately 10%, 20%, and 30% body weight for our sample. Main outcome measures were disc compression, defined as post- minus preloading disc height, and lumbar asymmetry, defined as the coronal Cobb angle between the superior endplates of S1 and L1. RESULTS: Increasing backpack loads significantly compressed lumbar disc heights measured in the midline sagittal plane (P < 0.05, repeated-measures analysis of variance [ANOVA]). Lumbar asymmetry was: 2.23 degrees +/- 1.07 degrees standing, 5.46 degrees +/- 2.50 degrees with 4 kg, 9.18 degrees +/- 2.25 degrees with 8 kg, and 5.68 degrees +/- 1.76 degrees with 12 kg (mean +/- SE). Backpack loads significantly increased lumbar asymmetry (P < 0.03, one-way ANOVA). Four of the 8 subjects had Cobb angles greater than 10 degrees during 8-kg backpack loads. Using a visual analogue scale to rate their pain (0-no pain, 10-worst pain imaginable), subjects reported significant increases in back pain associated with backpack loads of 4, 8, and 12 kg (P < 0.001, 1-way ANOVA). CONCLUSION: Backpack loads are responsible for a significant amount of back pain in children, which in part, may be due to changes in lumbar disc height or curvature. This is the first upright MRI study to document reduced disc height and greater lumbar asymmetry for common backpack loads in children. PMID- 20023609 TI - Should we screen for oral cancer? PMID- 20023608 TI - Prevalence of autism spectrum disorders - Autism and Developmental Disabilities Monitoring Network, United States, 2006. AB - PROBLEM/CONDITION: Autism spectrum disorders (ASDs) are a group of developmental disabilities characterized by atypical development in socialization, communication, and behavior. ASDs typically are apparent before age 3 years, with associated impairments affecting multiple areas of a person's life. Because no biologic marker exists for ASDs, identification is made by professionals who evaluate a child's developmental progress to identify the presence of developmental disorders. REPORTING PERIOD: 2006. METHODS: Earlier surveillance efforts indicated that age 8 years is a reasonable index age at which to monitor peak prevalence. The identified prevalence of ASDs in U.S. children aged 8 years was estimated through a systematic retrospective review of evaluation records in multiple sites participating in the Autism and Developmental Disabilities Monitoring (ADDM) Network. Data were collected from existing records in 11 ADDM Network sites (areas of Alabama, Arizona, Colorado, Florida, Georgia, Maryland, Missouri, North Carolina, Pennsylvania, South Carolina, and Wisconsin) for 2006. To analyze changes in identified ASD prevalence, CDC compared the 2006 data with data collected from 10 sites (all sites noted above except Florida) in 2002. Children aged 8 years with a notation of an ASD or descriptions consistent with an ASD were identified through screening and abstraction of existing health and education records containing professional assessments of the child's developmental progress at health-care or education facilities. Children aged 8 years whose parent(s) or legal guardian(s) resided in the respective areas in 2006 met the case definition for an ASD if their records documented behaviors consistent with the Diagnostic and Statistical Manual of Mental Disorders, 4th edition, text revision (DSM-IV-TR) criteria for autistic disorder, pervasive developmental disorder--not otherwise specified (PDD NOS), or Asperger disorder. Presence of an identified ASD was determined through a review of data abstracted from developmental evaluation records by trained clinician reviewers. RESULTS: For the 2006 surveillance year, 2,757 (0.9%) of 308,038 [corrected] children aged 8 years residing in the 11 ADDM sites were identified as having an ASD, indicating an overall average prevalence of 9.0 per 1,000 population (95% confidence interval [CI] = 8.6--9.3). ASD prevalence per 1,000 children aged 8 years ranged from 4.2 in Florida to 12.1 in Arizona and Missouri, with prevalence for the majority of sites ranging between 7.6 and 10.4. For 2006, ASD prevalence was significantly lower in Florida (p<0.001) and Alabama (p<0.05) and higher in Arizona and Missouri (p<0.05) than in all other sites. The ratio of males to females ranged from 3.2:1 in Alabama to 7.6:1 in Florida. ASD prevalence varied by type of ascertainment source, with higher average prevalence in sites with access to health and education records (10.0) compared with sites with health records only (7.5). Although parental or professional concerns regarding development before age 36 months were noted in the evaluation records of the majority of children who were identified as having an ASD, the median age of earliest documented ASD diagnosis was much later (range: 41 months [Florida]-60 months [Colorado]). Of 10 sites that collected data for both the 2002 and 2006 surveillance years, nine observed an increase in ASD prevalence (range: 27%-95% increase; p<0.01), with increases among males in all sites and among females in four of 11 sites, and variation among other subgroups. INTERPRETATION: In 2006, on average, approximately 1% or one child in every 110 in the 11 ADDM sites was classified as having an ASD (approximate range: 1:80-1:240 children [males: 1:70; females: 1:315]). The average prevalence of ASDs identified among children aged 8 years increased 57% in 10 sites from the 2002 to the 2006 ADDM surveillance year. Although improved ascertainment accounts for some of the prevalence increases documented in the ADDM sites, a true increase in the risk for children to develop ASD symptoms cannot be ruled out. On average, although delays in identification persisted, ASDs were being diagnosed by community professionals at earlier ages in 2006 than in 2002. PUBLIC HEALTH ACTIONS: These results indicate an increased prevalence of identified ASDs among U.S. children aged 8 years and underscore the need to regard ASDs as an urgent public health concern. Continued monitoring is needed to document and understand changes over time, including the multiple ascertainment and potential risk factors likely to be contributing. Research is needed to ascertain the factors that put certain persons at risk, and concerted efforts are essential to provide support for persons with ASDs, their families, and communities to improve long-term outcome. PMID- 20023610 TI - Oral health promotion initiated during pregnancy successful in reducing early childhood caries. AB - DESIGN: This was a randomised controlled trial using Zelen's design. INTERVENTION: Women in the test group received three rounds of printed information supplied in the form of anticipatory guidance. The first round of information was provided at enrolment. The second and the third rounds of information were mailed to the mother's home address when their child reached 6 and 12 months of age. The topics covered in each round offered mothers information and support relevant to their own and the child's oral development across the subsequent 6 months period. In addition, the first round of literature included information for expectant mothers related to oral health changes during pregnancy, with emphasis on oral hygiene during pregnancy and proper nutrition. Other topics covered in this round were the importance of primary teeth, the use of pacifiers and the sleeping patterns of the child. The second and third rounds of information were concerned with the eruption of teeth, oral hygiene and nutrition. The oral health promotion material was supplemented with the existing nutritional recommendations. Half of the mothers in the test group were randomly selected to test the impact of a structured telephone consultation as an additional mode of contact. The structured telephone consultation was conducted as a scripted interview. Four questionnaires were also conducted (a maternal oral health survey, refusal to participate, child's oral health survey, and evaluation of oral health intervention). OUTCOME MEASURES: The main outcome was severe early childhood caries (S-ECC). A case of S-ECC was defined when one or more upper incisor teeth labial surfaces were carious, either noncavitated or cavitated. Diagnosis was based on visual criteria only. RESULTS: Out of 649 women enrolled in the programme (test group, 327l control group, 322), 441 had their child examined at followup. The incidence of S-ECC in the test group was 1.7% and in the control group was 9.6% (P < 0.001). CONCLUSIONS: An oral health promotion programme based on repeated rounds of anticipatory guidance initiated during the mother's pregnancy was successful in reducing the incidence of S-ECC in these very young children. PMID- 20023611 TI - Prophylactic extraction of deciduous canines. AB - DATA SOURCES: The Cochrane Oral Health Group Trials Register, Cochrane Central Register of Controlled Trials, Medline and Embase were searched, along with searches by hand of relevant journals, when these had not already been searched as part of the Cochrane Oral Health Groups handsearching programme. The references of relevant publications were checked for further studies. STUDY SELECTION: Trials were selected if they met the following criteria: they were randomised controlled trials (RCT) or quasi-RCT, involved the extraction of the primary maxillary canine and assessed eruption/ noneruption of the palatally displaced maxillary permanent canine. DATA EXTRACTION AND SYNTHESIS: Seven review authors independently, in duplicate, examined the studies found in the search. The primary outcome was the reported prevalence of eruption or noneruption of the ectopic permanent canine into the mouth following observation or intervention. Results were to be expressed as risk ratios for dichotomous outcomes with 95% confidence intervals and mean differences for continuous outcomes. Heterogeneity was to be investigated, including both clinical and methodological factors. RESULTS: Of 324 publications identified, 295 were excluded after reviewing the abstract. Full articles were obtained for the remaining 31, of which 19 were not reported in English and required translation. Three reports of two RCT were identified for possible inclusion in the review but the data in the publications were not presented in a usable form and the authors have been contacted for further details. CONCLUSIONS: There is currently no evidence to support the extraction of the deciduous maxillary canine to facilitate the eruption of the palatally ectopic maxillary permanent canine. Two RCT were identified but, unfortunately, because of deficiencies in reporting, they cannot be included in the review at the present time. PMID- 20023612 TI - Prescribing systemic antibiotics when replanting avulsed teeth. AB - DATA SOURCES: Four databases were searched for relevant citations: Ovid Medline, Cochrane Library, PubMed and ISI Web of Science. STUDY SELECTION: Studies were included if they were conducted on human permanent anterior teeth and they reported systematic antibiotic use and periodontal healing outcomes. Only English language papers were considered. DATA EXTRACTION AND SYNTHESIS: A meta-analysis was conducted using a random-effects model. Heterogeneity was assessed using Cochran Q test and I(2) statistic. RESULTS: Three studies were included in the meta-analyses involving, in total, 326 teeth: 247 (76%) of these were treated with systemic antibiotics, whereas 79 (24%) had no systemic antibiotic therapy following replantation. There was no statistically significant difference between prescribing or not prescribing antibiotics for acceptable periodontal healing without progressive root resorption (common odds ratio, 0.90; standard error of the mean, 0.29; 95% confidence intervals, 0.51-1.58). CONCLUSIONS: There was inconclusive evidence for an association between systemic antibiotic therapy and an increased likelihood of acceptable periodontal healing. Pending future research to the contrary, dentists are recommended to follow current guidelines in prescribing systemic antibiotic therapy when replanting avulsed teeth. PMID- 20023613 TI - Splinting duration for replanted avulsed teeth. AB - DATA SOURCES: Four databases were searched for relevant citations: Ovid Medline, Cochrane Library, PubMed and ISI Web of Science. STUDY SELECTION: studies were included in the review if they had been conducted on humans and addressed the management of traumatised permanent teeth. Only English language papers were considered. Final selection was based on relevance to the question, and availability of sufficient detail in the paper. DATA EXTRACTION AND SYNTHESIS: A qualitative summary of the studies identified is presented. RESULTS: The review was limited by the studies' small sample sizes, variability in study design methodology and observation periods, and lack of uniformity in terminology for outcomes. A total of 138 replanted avulsed permanent teeth could be pooled from four papers, each reporting both short-term splinting (14 days or less) and long term splinting (over 14 days) in accord with current clinical guidelines. CONCLUSIONS: The evidence appears inconclusive for any association between short term splinting and increased likelihood of functional periodontal healing, acceptable healing, or decreased development of replacement resorption. The study found no evidence to contraindicate the current guidelines and suggests that the likelihood of successful periodontal healing after replantation is unaffected by splinting duration. Pending future research to the contrary, it is recommended that dentists continue to use the currently-recommended splinting periods when replanting avulsed permanent teeth. PMID- 20023614 TI - Periodontal treatment during pregnancy did not reduce the occurrence of poor pregnancy outcomes. AB - DESIGN: This was a randomised controlled trial (RCT). INTERVENTION: Participants were assigned randomly to receive periodontal treatment either before 23 weeks' gestational age or after delivery. Treatment consisted of up to four sessions of supragingival and subgingival scaling and root planing using hand and ultrasonic instruments to complete the baseline therapy. Local anaesthesia was used as needed. Those in the treatment group also received full-mouth tooth polishing and oral hygiene home instructions. There were no followup periodontal treatment visits. After delivery, those in the delayed-treatment group received periodontal therapy at no cost. OUTCOME MEASURE: The principal outcome (gestational age <37 weeks) included induced or spontaneous deliveries, foetal demise and miscarriage, but not therapeutic abortions. Secondary outcomes included gestational age <35 weeks; mean birth weight among neonates adjusted for race, sex and gestational age; and a composite measure of neonatal morbidity and mortality before discharge. RESULTS: The trial randomised 1806 patients at three performance sites and 1760 evaluable patients completed the study. At baseline, there were no differences between the treatment and control arms for any of the periodontal or obstetric measures. The rate of preterm delivery for the treatment group was 13.1%, and 11.5% for the control group (P 0.316). There were no significant differences when comparing women in the treatment group with those in the control group with regard to the adverse event rate or the major obstetric and neonatal outcomes. CONCLUSIONS: Periodontal therapy did not reduce the incidence of preterm delivery. PMID- 20023615 TI - No available evidence to assess the effectiveness of bonded amalgams. AB - DATA SOURCES: Sources of data were the Cochrane Oral Health Group's Trials Register, Cochrane CENTRAL (Cochrane Library), Medline and Embase. Reference lists of relevant articles were also searched and the investigators of the included studies were contacted for details of additional published and unpublished trials. STUDY SELECTION: Randomised controlled trials (RCT) were chosen that compared adhesively bonded versus traditional nonbonded amalgam restorations in conventional preparations utilising deliberate retention, in adults with permanent molar and premolar teeth suitable for Class I and II amalgam restorations only. DATA EXTRACTION AND SYNTHESIS: Two review authors independently screened papers, extracted trial details and assessed the risk of bias in the included study. RESULTS: One trial was included, comprising 31 patients who received 113 restorations. At 2 years, only three out of 53 restorations in the nonbonded group were lost, which was attributed to a lack of retention, and 55 out of 60 bonded restorations survived, with five unaccounted for at followup. Postinsertion sensitivity was not significantly different (P >0.05) at baseline or 2-year followup. No fractures of tooth tissue were reported and there was no significant difference between the groups or matched pairs of restorations in their marginal adaptation (P >0.05). CONCLUSIONS: There is no evidence to either claim or refute a difference in survival between bonded and nonbonded amalgam restorations. This review only found one methodologically sound but somewhat under-reported trial. This trial did not find any significant difference in the in-service performance of moderately sized adhesively bonded amalgam restorations, in terms of their survival rate and marginal integrity, in comparison with nonbonded amalgam restorations over a 2-year period. In view of the lack of evidence on the additional benefit of adhesively bonding amalgam compared with with nonbonded amalgam, it is important that clinicians are mindful of the additional costs that may be incurred. PMID- 20023616 TI - Fine-hybrid and nanohybrid restorative materials show similar durability in Class II cavities. AB - DESIGN: This was a randomised trial. INTERVENTION: Individuals who required a minimum of two replacement fillings were recruited. Restorations were placed using either Grandio bonded with Solobond M (Voco, Cuxhaven, Germany) or Tetric Ceram bonded with Syntac (Ivoclar Vivadent, Schaan, Liechtenstein). OUTCOME MEASURE: At the initial recall (baseline, ie, within 2 weeks), and after 6 months, 1 and 2 years, all restorations were assessed according to the modified United States Public Health Service (US PHS) criteria by two independent investigators using mirrors, probes, bitewing radiographs, impressions and intra oral photographs. Recall assessments were not performed by the clinician who initially placed the restorations. RESULTS: Both recall rate and survival rate were 100% after 4 years of clinical service. No significant difference was found between the restorative materials. Hypersensitivities were significantly reduced over time (P < 0.05; Friedman test). A significant deterioration over time was found for the criteria of marginal integrity (66% sufficient after 4 years),tooth integrity (15% sufficient), filling integrity (73% sufficient) and proximal contact. Stereo light microscopy and scanning electron microscopy (SEM) analysis of restoration margins revealed differences in the amount of perfect margins in favour of Tetric Ceram (P <0.05). CONCLUSIONS: Both materials performed satisfactorily over the 4-year observation period. Because of the extension of the restorations, wear was clearly visible after 4 years of clinical service with 50% sufficient ratings. PMID- 20023617 TI - Lack of evidence about the effectiveness of the different denture cleaning methods. AB - DATA SOURCES: Relevant studies were sourced by searching the Cochrane Oral Health Group Trials Register, Cochrane CENTRAL (Cochrane Library), Medline, Embase, LILACS and CINAHL. There were no language restrictions. STUDY SELECTION: Randomised controlled trials (RCT) were considered if they compared any mechanical method (eg, brushing or ultrasound) or chemical (eg, enzymes, sodium hypochlorite, oral rinses or peroxide solutions) in adults over the age of 18 years who wore removable partial dentures or complete dentures. DATA EXTRACTION AND SYNTHESIS: Two independent review authors screened and extracted information from, and independently assessed the risk of bias in, the included trials. RESULTS: Although six RCT were included in this review, the wide range of different interventions and outcome variables did not permit pooling of data in a meta-analysis. Isolated reports indicated that chemicals and brushing appear to be more effective than placebo in reducing plaque coverage and reducing microbial counts of anaerobes and aerobes on complete denture bases. CONCLUSIONS: There is a lack of evidence about the comparative effectiveness of the different denture cleaning methods considered in this review. Few well-designed RCT were found. Future research should focus on comparisons between mechanical and chemical methods, should assess the association between methods, and consider primary variables and costs. PMID- 20023618 TI - Temporomandibular joint arthrocentesis and lavage. AB - DATA SOURCES: The Cochrane Oral Health Group's Trials Register, Cochrane CENTRAL (Cochrane Library), Medline and Embase were used to identify likely studies. OpenSIGLE,(*) CBMdisc (the Chinese Biomedical Literature Database) and the Chinese Medical Library were also searched. All the Chinese professional journals in the oral health field were searched by hand and conference proceedings were consulted. There was no language restriction. STUDY SELECTION: Randomised controlled trials (RCT) and quasi-RCT that aimed to test the therapeutic effects of arthrocentesis and lavage on temporomandibular joint (TMJ) disorders were included.ATA EXTRACTION AND SYNTHESIS: Two review authors independently extracted data, and three review authors independently assessed the risk of bias of included trials. The first authors of the selected articles were contacted for additional information. RESULTS: Two trials were included in the review, one of which was judged to have a high risk of bias and one to be unclear in its risk of bias. The two trials, which included 81 individuals who had TMJ disorders, compared arthrocentesis with arthroscopy. No statistically significant difference was found between the interventions in terms of pain. A statistically significant difference was found in favour of arthroscopy in maximum incisal opening (weighted mean difference of -5.28; 95% confidence interval, -7.10 to -3.46). Mild and transient adverse reactions, such as discomfort or pain at the injection site, were reported in both groups. No data about quality of life were reported. CONCLUSIONS: There is insufficient, consistent evidence to either support or refute the use of arthrocentesis and lavage for treating patients with TMJ disorders. Further high-quality RCT of arthrocentesis need to be conducted before firm conclusions can be drawn with regard to its effectiveness. PMID- 20023619 TI - Oral health promotion programmes during orthodontic treatment beneficial. AB - DATA SOURCES: The Cochrane Central Register of Controlled Trials, Medline and Embase databases were searched. A hand search was made of the American Journal of Orthodontics and Dentofacial Orthopaedics, (British) Journal of Orthodontics, European Journal of Orthodontics and Angle Orthodontist, Google Scholar and the reference lists of relevant articles. STUDY SELECTION: Only randomised controlled trials (RCT) and quasi-randomised controlled clinical trials (CCT), which specifically stated that they assessed reductions in dental plaque levels and/ or gingival bleeding when comparing oral health promotion (OHP) interventions, were included. Trials that involved plaque removal by a professional (except at baseline) or the use of proprietary antiplaque agents were excluded. DATA EXTRACTION AND SYNTHESIS: Data extraction was carried out independently by two reviewers. Study quality was assessed for their method of allocation, concealment of allocation, masking of assessment and reporting of withdrawals. Direct comparison between the trials was difficult because of the heterogeneity in the outcome measures between the included studies. RESULTS: Six RCT and quasi randomised CCT met the inclusion criteria. Positive effects on plaque and/ or gingival health were produced in only four of the included trials. OHP did not result in any detectable difference in two of the included trials. None of the trials that were included produced a negative effect of orthodontic OHP on oral hygiene and gingival health. CONCLUSIONS: An OHP programme for people undergoing fixed appliance orthodontic treatment produces a short-term reduction (of up to 5 months) in plaque and improvement in gingival health. No particular OHP method produced a greater short-term benefit to periodontal health during fixed appliance orthodontic treatment. Further studies using appropriate methods and, in particular,r longer followup periods are required. PMID- 20023620 TI - Mini-implants for direct or indirect orthodontic anchorage. AB - DATA SOURCES: Studies were sourced using Medline, the Cochrane Library and hand searches of key orthodontic journals. STUDY SELECTION: The review was restricted to peer-review articles (randomised controlled studies, prospective clinical studies and retrospective clinical studies) dealing with mini-implants (implant diameter smaller than 2.5 mm) and conducted in humans. Articles were excluded if: they dealt with standard dental implants, onplants (palatal implants), miniplates used as orthodontic anchorage, or miniscrews or microscrews for dental surgery, and implant materials research; if they were animal studies, in-vitro studies, case reports and case series, or technique presentations of mini-implants and micro-implants, review articles and letters; or if they were articles that did not meet the objective of this review or were reported in a language other than English. DATA EXTRACTION AND SYNTHESIS: Data were extracted independently by two authors, with disagreements resolved by discussion. Study quality was assessed. A qualitative synthesis was conducted. RESULTS: Sixteen articles met the inclusion criteria and, because of quality limitations, the literature was summarised in two broad areas, placement-related and loading-related factors. CONCLUSIONS: Mini implants are effective as anchorage, and their success depends on proper initial mechanical stability and loading quality and quantity. PMID- 20023621 TI - Vitamin B12 for the treatment of recurrent aphthous stomatitis. AB - DESIGN: This was a randomised, double-blind, placebo-controlled trial. INTERVENTION: One tablet was taken each day before sleep for 6 months. The test group received sublingual vitamin B12 tablets (1000 mcg of vitamin B12) whereas the control group took a placebo of the same shape, size, colour and flavour. Participants met with staff monthly. OUTCOME MEASURE: Duration (days) of an aphthous stomatitis episode, monthly number of aphthous ulcers, and severity of pain according to the Numerous Rating Scale (NRS), were recorded in a diary. RESULTS: Fifty-eight people suffering from recurrent aphthous stomatitis (RAS) participated: 31 were allocated to the intervention group and 27 to the control group. The duration of outbreaks, the number of ulcers, and the level of pain were reduced significantly (P <0.05) at 5 and 6 months of treatment with vitamin B12, regardless of initial vitamin B12 levels in the blood. During the last month of treatment a significant number of participants in the intervention group reached "no aphthous ulcers status" (74.1% vs 32.0%; P <0.01). CONCLUSIONS: Vitamin B12 treatment, which is simple, inexpensive and low-risk, seems to be effective for patients suffering from RAS, regardless of the serum vitamin B12 level. PMID- 20023622 TI - Interventions for prevention and treatment of herpes simplex virus in cancer patients. AB - DATA SOURCES: Relevant data was sourced using the Cochrane Oral Health Group Trials Register, Cochrane Central Register of Controlled Trials, Medline, Embase, CINAHL, CANCERLIT, SIGLE and LILACS. Searching by hand was also carried out, reference lists checked for further trials, and authors and known specialists in the field contacted to try to identify any additional published or unpublished trials. STUDY SELECTION: Randomised controlled trials that concerned treatment or prophylaxis of orofacial lesions [caused by herpes simplex virus (HSV)] in adults, children (or both) who were immunocompromised because of cancer were eligible for inclusion. Outcomes of interest were presence/ absence of clinical/ culture-positive HSV infections (prevention), time to complete healing of lesions (treatment), duration of viral shedding, recurrence of lesions, relief of pain, amount of analgesia, duration of hospital stay, cost of oral care, patient quality of life, and adverse effects. The reports obtained from the electronic and other forms of searches were assessed independently by the review authors. Disagreements were resolved by discussion and reasons recorded. DATA EXTRACTION AND SYNTHESIS: Authors were contacted for details of randomisation, blindness and sample demographics. Quality assessment was carried out on randomisation, blindness, withdrawals and selective reporting. The Cochrane Collaborations statistical guidelines were followed and risk ratio values were calculated using random effects models. RESULTS: In the 17 trials, three interventions were studied: use of aciclovir, valaciclovir and prostaglandin E. No trials reported on duration of hospital stay, amount of analgesia or patient quality of life. In the placebo-controlled trials, aciclovir was found to be effective for prevention and treatment. In comparisons of active interventions, there is no evidence of a significant difference in efficacy between valaciclovir and aciclovir, or higher doses of valaciclovir and lower doses. In the single study assessing the effectiveness of prostaglandin E for prevention, this intervention was less effective than placebo. No adverse effects were reported. CONCLUSIONS: There is evidence that aciclovir is effective at preventing and treating HSV infections. There is no evidence that valaciclovir is more efficacious than aciclovir, or that a high dose of valaciclovir is better than a low dose of valaciclovir. There is evidence that, as a prophylaxis, placebo is more efficacious than prostaglandin E. In all included trials the risk of bias was unclear. PMID- 20023623 TI - NHS clinical knowledge summaries. AB - The UK National Health Service (NHS) Clinical Knowledge Summaries, formerly known as PRODIGY, are part of the National Library for Health and provide a source of evidence-based information and practical know-how relating to the common conditions managed in primary care. PMID- 20023627 TI - A clean slate. PMID- 20023628 TI - Crystal structure of DNA-PKcs reveals a large open-ring cradle comprised of HEAT repeats. AB - Broken chromosomes arising from DNA double-strand breaks result from endogenous events such as the production of reactive oxygen species during cellular metabolism, as well as from exogenous sources such as ionizing radiation. Left unrepaired or incorrectly repaired they can lead to genomic changes that may result in cell death or cancer. DNA-dependent protein kinase (DNA-PK), a holoenzyme that comprises the DNA-PK catalytic subunit (DNA-PKcs) and the heterodimer Ku70/Ku80, has a major role in non-homologous end joining-the main pathway in mammals used to repair double-strand breaks. DNA-PKcs is a serine/threonine protein kinase comprising a single polypeptide chain of 4,128 amino acids and belonging to the phosphatidylinositol-3-OH kinase (PI(3)K) related protein family. DNA-PKcs is involved in the sensing and transmission of DNA damage signals to proteins such as p53, setting off events that lead to cell cycle arrest. It phosphorylates a wide range of substrates in vitro, including Ku70/Ku80, which is translocated along DNA. Here we present the crystal structure of human DNA-PKcs at 6.6 A resolution, in which the overall fold is clearly visible, to our knowledge, for the first time. The many alpha-helical HEAT repeats (helix-turn-helix motifs) facilitate bending and allow the polypeptide chain to fold into a hollow circular structure. The carboxy-terminal kinase domain is located on top of this structure, and a small HEAT repeat domain that probably binds DNA is inside. The structure provides a flexible cradle to promote DNA double-strand-break repair. PMID- 20023629 TI - Deubiquitinase USP9X stabilizes MCL1 and promotes tumour cell survival. AB - MCL1 is essential for the survival of stem and progenitor cells of multiple lineages, and is unique among pro-survival BCL2 family members in that it is rapidly turned over through the action of ubiquitin ligases. B- and mantle-cell lymphomas, chronic myeloid leukaemia, and multiple myeloma, however, express abnormally high levels of MCL1, contributing to chemoresistance and disease relapse. The mechanism of MCL1 overexpression in cancer is not well understood. Here we show that the deubiquitinase USP9X stabilizes MCL1 and thereby promotes cell survival. USP9X binds MCL1 and removes the Lys 48-linked polyubiquitin chains that normally mark MCL1 for proteasomal degradation. Increased USP9X expression correlates with increased MCL1 protein in human follicular lymphomas and diffuse large B-cell lymphomas. Moreover, patients with multiple myeloma overexpressing USP9X have a poor prognosis. Knockdown of USP9X increases MCL1 polyubiquitination, which enhances MCL1 turnover and cell killing by the BH3 mimetic ABT-737. These results identify USP9X as a prognostic and therapeutic target, and they show that deubiquitinases may stabilize labile oncoproteins in human malignancies. PMID- 20023630 TI - Innate production of T(H)2 cytokines by adipose tissue-associated c-Kit(+)Sca 1(+) lymphoid cells. AB - Innate immune responses are important in combating various microbes during the early phases of infection. Natural killer (NK) cells are innate lymphocytes that, unlike T and B lymphocytes, do not express antigen receptors but rapidly exhibit cytotoxic activities against virus-infected cells and produce various cytokines. Here we report a new type of innate lymphocyte present in a novel lymphoid structure associated with adipose tissues in the peritoneal cavity. These cells do not express lineage (Lin) markers but do express c-Kit, Sca-1 (also known as Ly6a), IL7R and IL33R. Similar lymphoid clusters were found in both human and mouse mesentery and we term this tissue 'FALC' (fat-associated lymphoid cluster). FALC Lin(-)c-Kit(+)Sca-1(+) cells are distinct from lymphoid progenitors and lymphoid tissue inducer cells. These cells proliferate in response to IL2 and produce large amounts of T(H)2 cytokines such as IL5, IL6 and IL13. IL5 and IL6 regulate B-cell antibody production and self-renewal of B1 cells. Indeed, FALC Lin(-)c-Kit(+)Sca-1(+) cells support the self-renewal of B1 cells and enhance IgA production. IL5 and IL13 mediate allergic inflammation and protection against helminth infection. After helminth infection and in response to IL33, FALC Lin( )c-Kit(+)Sca-1(+) cells produce large amounts of IL13, which leads to goblet cell hyperplasia-a critical step for helminth expulsion. In mice devoid of FALC Lin( )c-Kit(+)Sca-1(+) cells, such goblet cell hyperplasia was not induced. Thus, FALC Lin(-)c-Kit(+)Sca-1(+) cells are T(H)2-type innate lymphocytes, and we propose that these cells be called 'natural helper cells'. PMID- 20023631 TI - Extreme-angle broadband metamaterial lens. AB - For centuries, the conventional approach to lens design has been to grind the surfaces of a uniform material in such a manner as to sculpt the paths that rays of light follow as they transit through the interfaces. Refractive lenses formed by this procedure of bending the surfaces can be of extremely high quality, but are nevertheless limited by geometrical and wave aberrations that are inherent to the manner in which light refracts at the interface between two materials. Conceptually, a more natural--but usually less convenient--approach to lens design would be to vary the refractive index throughout an entire volume of space. In this manner, far greater control can be achieved over the ray trajectories. Here, we demonstrate how powerful emerging techniques in the field of transformation optics can be used to harness the flexibility of gradient index materials for imaging applications. In particular we design and experimentally demonstrate a lens that is broadband (more than a full decade bandwidth), has a field-of-view approaching 180 degrees and zero f-number. Measurements on a metamaterial implementation of the lens illustrate the practicality of transformation optics to achieve a new class of optical devices. PMID- 20023632 TI - Three-dimensional imaging of strain in a single ZnO nanorod. AB - Nanoscale structures can be highly strained because of confinement effects and the strong influence of their external boundaries. This results in dramatically different electronic, magnetic and optical material properties of considerable utility. Third-generation synchrotron-based coherent X-ray diffraction has emerged as a non-destructive tool for three-dimensional (3D) imaging of strain and defects in crystals that are smaller than the coherence volume, typically a few cubic micrometres, of the available beams that have sufficient flux to reveal the material's structure. Until now, measurements have been possible only at a single Bragg point of a given crystal because of the limited ability to maintain alignment; it has therefore been possible to determine only one component of displacement and not the full strain tensor. Here we report key advances in our fabrication and experimental techniques, which have enabled diffraction patterns to be obtained from six Bragg reflections of the same ZnO nanocrystal for the first time. All three Cartesian components of the ion displacement field, and in turn the full nine-component strain tensor, have thereby been imaged in three dimensions. PMID- 20023633 TI - Dynamic display of biomolecular patterns through an elastic creasing instability of stimuli-responsive hydrogels. AB - Surfaces with physicochemical properties that can be modulated using external stimuli offer great promise for designing responsive or adaptive materials. Here, we describe biocompatible dynamic scaffolds based on thin hydrogel coatings that reversibly hide and display surface chemical patterns in response to temperature changes. At room temperature, the gel absorbs water, triggering an elastic creasing instability that sequesters functionalized regions within tight folds in the surface. Deswelling at approximately 37 degrees C causes the gel surface to unfold, thereby regenerating the biomolecular patterns. Crease positions are directed by topographic features on the underlying substrate, and are translated into two-dimensional micrometre-scale surface chemical patterns through selective deposition of biochemically functionalized polyelectrolytes. We demonstrate specific applications of these dynamic scaffolds--selective capture, sequestration and release of micrometre-sized beads, tunable activity of surface immobilized enzymes and reversible encapsulation of adherent cells--which offer promise for incorporation within lab-on-a-chip devices or as dynamic substrates for cellular biology. PMID- 20023634 TI - Real-time imaging reveals the single steps of brain metastasis formation. AB - Brain metastasis frequently occurs in individuals with cancer and is often fatal. We used multiphoton laser scanning microscopy to image the single steps of metastasis formation in real time. Thus, it was possible to track the fate of individual metastasizing cancer cells in vivo in relation to blood vessels deep in the mouse brain over minutes to months. The essential steps in this model were arrest at vascular branch points, early extravasation, persistent close contacts to microvessels and perivascular growth by vessel cooption (melanoma) or early angiogenesis (lung cancer). Inefficient steps differed between the tumor types. Long-term dormancy was only observed for single perivascular cancer cells, some of which moved continuously. Vascular endothelial growth factor-A (VEGF-A) inhibition induced long-term dormancy of lung cancer micrometastases by preventing angiogenic growth to macrometastases. The ability to image the establishment of brain metastases in vivo provides new insights into their evolution and response to therapies. PMID- 20023635 TI - Generation of stable monoclonal antibody-producing B cell receptor-positive human memory B cells by genetic programming. AB - The B cell lymphoma-6 (Bcl-6) and Bcl-xL proteins are expressed in germinal center B cells and enable them to endure the proliferative and mutagenic environment of the germinal center. By introducing these genes into peripheral blood memory B cells and culturing these cells with two factors produced by follicular helper T cells, CD40 ligand (CD40L) and interleukin-21 (IL-21), we convert them to highly proliferating, cell surface B cell receptor (BCR) positive, immunoglobulin-secreting B cells with features of germinal center B cells, including expression of activation-induced cytidine deaminase (AID). We generated cloned lines of B cells specific for respiratory syncytial virus and used these cells as a source of antibodies that effectively neutralized this virus in vivo. This method provides a new tool to study B cell biology and signal transduction through antigen-specific B cell receptors and for the rapid generation of high-affinity human monoclonal antibodies. PMID- 20023636 TI - Biofilm-like extracellular viral assemblies mediate HTLV-1 cell-to-cell transmission at virological synapses. AB - Human T cell leukemia virus type 1 (HTLV-1) is a lymphotropic retrovirus whose cell-to-cell transmission requires cell contacts. HTLV-1-infected T lymphocytes form 'virological synapses', but the mechanism of HTLV-1 transmission remains poorly understood. We show here that HTLV-1-infected T lymphocytes transiently store viral particles as carbohydrate-rich extracellular assemblies that are held together and attached to the cell surface by virally-induced extracellular matrix components, including collagen and agrin, and cellular linker proteins, such as tetherin and galectin-3. Extracellular viral assemblies rapidly adhere to other cells upon cell contact, allowing virus spread and infection of target cells. Their removal strongly reduces the ability of HTLV-1-producing cells to infect target cells. Our findings unveil a novel virus transmission mechanism based on the generation of extracellular viral particle assemblies whose structure, composition and function resemble those of bacterial biofilms. HTLV-1 biofilm like structures represent a major route for virus transmission from cell to cell. PMID- 20023637 TI - Salt-sensitive hypertension in circadian clock-deficient Cry-null mice involves dysregulated adrenal Hsd3b6. AB - Malfunction of the circadian clock has been linked to the pathogenesis of a variety of diseases. We show that mice lacking the core clock components Cryptochrome-1 (Cry1) and Cryptochrome-2 (Cry2) (Cry-null mice) show salt sensitive hypertension due to abnormally high synthesis of the mineralocorticoid aldosterone by the adrenal gland. An extensive search for the underlying cause led us to identify type VI 3beta-hydroxyl-steroid dehydrogenase (Hsd3b6) as a new hypertension risk factor in mice. Hsd3b6 is expressed exclusively in aldosterone producing cells and is under transcriptional control of the circadian clock. In Cry-null mice, Hsd3b6 messenger RNA and protein levels are constitutively high, leading to a marked increase in 3beta-hydroxysteroid dehydrogenase-isomerase (3beta-HSD) enzymatic activity and, as a consequence, enhanced aldosterone production. These data place Hsd3b6 in a pivotal position through which circadian clock malfunction is coupled to the development of hypertension. Translation of these findings to humans will require clinical examination of human HSD3B1 gene, which we found to be functionally similar to mouse Hsd3b6. PMID- 20023638 TI - Enzymatic and structural insights for substrate specificity of a family of jumonji histone lysine demethylases. AB - Combinatorial readout of multiple covalent histone modifications is poorly understood. We provide insights into how an activating histone mark, in combination with linked repressive marks, is differentially 'read' by two related human demethylases, PHF8 and KIAA1718 (also known as JHDM1D). Both enzymes harbor a plant homeodomain (PHD) that binds Lys4-trimethylated histone 3 (H3K4me3) and a jumonji domain that demethylates either H3K9me2 or H3K27me2. The presence of H3K4me3 on the same peptide as H3K9me2 makes the doubly methylated peptide a markedly better substrate of PHF8, whereas the presence of H3K4me3 has the opposite effect, diminishing the H3K9me2 demethylase activity of KIAA1718 without adversely affecting its H3K27me2 activity. The difference in substrate specificity between the two is explained by PHF8 adopting a bent conformation, allowing each of its domains to engage its respective target, whereas KIAA1718 adopts an extended conformation, which prevents its access to H3K9me2 by its jumonji domain when its PHD engages H3K4me3. PMID- 20023639 TI - Strong cooperativity between subunits in voltage-gated proton channels. AB - Voltage-activated proton (Hv) channels are essential components in the innate immune response. Hv channels are dimeric proteins with one proton permeation pathway per subunit. It is unknown how Hv channels are activated by voltage and whether there is any cooperation between subunits during voltage activation. Using cysteine accessibility measurements and voltage-clamp fluorometry, we show data consistent with the possibility that the fourth transmembrane segment S4 functions as the voltage sensor in Ciona intestinalis Hv channels. Unexpectedly, in a dimeric Hv channel, the S4 in both subunits must move to activate the two proton permeation pathways. In contrast, if Hv subunits are prevented from dimerizing, the movement of a single S4 is sufficient to activate the proton permeation pathway in a subunit. These results indicate strong cooperativity between subunits in dimeric Hv channels. PMID- 20023640 TI - The opening of the two pores of the Hv1 voltage-gated proton channel is tuned by cooperativity. AB - In voltage-gated sodium, potassium and calcium channels, the functions of ion conduction and voltage sensing are performed by two distinct structural units: the pore domain and the voltage-sensing domain (VSD). In the hydrogen voltage gated channel 1 (Hv1), the VSD, unusually, performs both functions. Hv1 was recently found to dimerize and to form channels made of two pores. However, the channels were also found to function when dimerization was prevented, raising a question about the functional role of dimerization. Here we show that the two subunits of the human Hv1 dimer influence one another during gating, with positive cooperativity shaping the response to voltage of the two pores. We also find that the two voltage sensors undergo conformational changes that precede pore opening and that these conformational changes are allosterically coupled between the two subunits. Our results point to an important role for dimerization in the modulation of Hv1 activity. PMID- 20023641 TI - An intramembrane aromatic network determines pentameric assembly of Cys-loop receptors. AB - Cys-loop receptors are pentameric ligand-gated ion channels (pLGICs) that mediate fast synaptic transmission. Here functional pentameric assembly of truncated fragments comprising the ligand-binding N-terminal ectodomains and the first three transmembrane helices, M1-M3, of both the inhibitory glycine receptor (GlyR) alpha1 and the 5HT(3)A receptor subunits was found to be rescued by coexpressing the complementary fourth transmembrane helix, M4. Alanine scanning identified multiple aromatic residues in M1, M3 and M4 as key determinants of GlyR assembly. Homology modeling and molecular dynamics simulations revealed that these residues define an interhelical aromatic network, which we propose determines the geometry of M1-M4 tetrahelical packing such that nascent pLGIC subunits must adopt a closed fivefold symmetry. Because pLGIC ectodomains form random nonstoichiometric oligomers, proper pentameric assembly apparently depends on intersubunit interactions between extracellular domains and intrasubunit interactions between transmembrane segments. PMID- 20023642 TI - New modes for subsurface atomic force microscopy through nanomechanical coupling. AB - Non-destructive, nanoscale characterization techniques are needed to understand both synthetic and biological materials. The atomic force microscope uses a force sensing cantilever with a sharp tip to measure the topography and other properties of surfaces. As the tip is scanned over the surface it experiences attractive and repulsive forces that depend on the chemical and mechanical properties of the sample. Here we show that an atomic force microscope can obtain a range of surface and subsurface information by making use of the nonlinear nanomechanical coupling between the probe and the sample. This technique, which is called mode-synthesizing atomic force microscopy, relies on multi-harmonic forcing of the sample and the probe. A rich spectrum of first- and higher-order couplings is discovered, providing a multitude of new operational modes for force microscopy, and the capabilities of the technique are demonstrated by examining nanofabricated samples and plant cells. PMID- 20023643 TI - Colloidal lenses allow high-temperature single-molecule imaging and improve fluorophore photostability. AB - Although single-molecule fluorescence spectroscopy was first demonstrated at near absolute zero temperatures (1.8 K), the field has since advanced to include room temperature observations, largely owing to the use of objective lenses with high numerical aperture, brighter fluorophores and more sensitive detectors. This has opened the door for many chemical and biological systems to be studied at native temperatures at the single-molecule level both in vitro and in vivo. However, it is difficult to study systems and phenomena at temperatures above 37 degrees C, because the index-matching fluids used with high-numerical-aperture objective lenses can conduct heat from the sample to the lens, and sustained exposure to high temperatures can cause the lens to fail. Here, we report that TiO(2) colloids with diameters of 2 microm and a high refractive index can act as lenses that are capable of single-molecule imaging at 70 degrees C when placed in immediate proximity to an emitting molecule. The optical system is completed by a low-numerical-aperture optic that can have a long working distance and an air interface, which allows the sample to be independently heated. Colloidal lenses were used for parallel imaging of surface-immobilized single fluorophores and for real-time single-molecule measurements of mesophilic and thermophilic enzymes at 70 degrees C. Fluorophores in close proximity to TiO(2) also showed a 40% increase in photostability due to a reduction of the excited-state lifetime. PMID- 20023644 TI - Large-area spatially ordered arrays of gold nanoparticles directed by lithographically confined DNA origami. AB - The development of nanoscale electronic and photonic devices will require a combination of the high throughput of lithographic patterning and the high resolution and chemical precision afforded by self-assembly. However, the incorporation of nanomaterials with dimensions of less than 10 nm into functional devices has been hindered by the disparity between their size and the 100 nm feature sizes that can be routinely generated by lithography. Biomolecules offer a bridge between the two size regimes, with sub-10 nm dimensions, synthetic flexibility and a capability for self-recognition. Here, we report the directed assembly of 5-nm gold particles into large-area, spatially ordered, two dimensional arrays through the site-selective deposition of mesoscopic DNA origami onto lithographically patterned substrates and the precise binding of gold nanocrystals to each DNA structure. We show organization with registry both within an individual DNA template and between components on neighbouring DNA origami, expanding the generality of this method towards many types of patterns and sizes. PMID- 20023645 TI - Electrostatic focusing of unlabelled DNA into nanoscale pores using a salt gradient. AB - Solid-state nanopores are sensors capable of analysing individual unlabelled DNA molecules in solution. Although the critical information obtained from nanopores (for example, DNA sequence) comes from the signal collected during DNA translocation, the throughput of the method is determined by the rate at which molecules arrive and thread into the pores. Here, we study the process of DNA capture into nanofabricated SiN pores of molecular dimensions. For fixed analyte concentrations we find an increase in capture rate as the DNA length increases from 800 to 8,000 base pairs, a length-independent capture rate for longer molecules, and increasing capture rates when ionic gradients are established across the pore. Furthermore, we show that application of a 20-fold salt gradient allows the detection of picomolar DNA concentrations at high throughput. The salt gradients enhance the electric field, focusing more molecules into the pore, thereby advancing the possibility of analysing unamplified DNA samples using nanopores. PMID- 20023646 TI - Switching binary states of nanoparticle superlattices and dimer clusters by DNA strands. AB - Nanoscale components can be self-assembled into static three-dimensional structures, arrays and clusters using biomolecular motifs. The structural plasticity of biomolecules and the reversibility of their interactions can also be used to make nanostructures that are dynamic, reconfigurable and responsive. DNA has emerged as an ideal biomolecular motif for making such nanostructures, partly because its versatile morphology permits in situ conformational changes using molecular stimuli. This has allowed DNA nanostructures to exhibit reconfigurable topologies and mechanical movement. Recently, researchers have begun to translate this approach to nanoparticle interfaces, where, for example, the distances between nanoparticles can be modulated, resulting in a distance dependent plasmonic response. Here, we report the assembly of nanoparticles into three-dimensional superlattices and dimer clusters, using a reconfigurable DNA device that acts as an interparticle linkage. The interparticle distances in the superlattices and clusters can be modified, while preserving structural integrity, by adding molecular stimuli (simple DNA strands) after the self assembly processes has been completed. Both systems were found to switch between two distinct rigid states, but a transition to a flexible device configuration within a superlattice showed a significant hysteresis. PMID- 20023647 TI - Electrically controlled DNA adhesion. AB - The ability to control the interaction of polyelectrolytes, such as DNA or proteins, with charged surfaces is of pivotal importance for a multitude of biotechnological applications. Previously, we measured the desorption forces of single polymers on charged surfaces using an atomic force microscope. Here, we show that the adhesion of DNA on gold electrodes modified with self-assembled monolayers can be biased by the composition of the monolayer and externally controlled by means of the electrode potential. Positive potentials induced DNA adsorption onto OH-terminated electrodes with adhesion forces up to 25 pN (at +0.5 V versus Ag/AgCl), whereas negative potentials suppressed DNA adsorption. The measured contributions of the DNA backbone phosphate charges and the doubly charged terminal phosphate on adsorption agreed with a model based on the Gouy Chapman theory. Experiments on an NH(2)-terminated electrode revealed a similar force modulation range of the coulomb component of the desorption force. These findings are important for the development of new DNA-based biochips or supramolecular structures. PMID- 20023648 TI - HERC2 coordinates ubiquitin-dependent assembly of DNA repair factors on damaged chromosomes. AB - Regulatory ubiquitylation is emerging as an important mechanism to protect genome integrity in cells exposed to DNA damage. However, the spectrum of known ubiquitin regulators of the DNA damage response (DDR) is limited and their functional interplay is poorly understood. Here, we identify HERC2 as a factor that regulates ubiquitin-dependent retention of repair proteins on damaged chromosomes. In response to ionising radiation (IR), HERC2 forms a complex with RNF8, a ubiquitin ligase involved in the DDR. The HERC2-RNF8 interaction requires IR-inducible phosphorylation of HERC2 at Thr 4827, which in turn binds to the forkhead-associated (FHA) domain of RNF8. Mechanistically, we provide evidence that HERC2 facilitates assembly of the ubiquitin-conjugating enzyme Ubc13 with RNF8, thereby promoting DNA damage-induced formation of Lys 63-linked ubiquitin chains. We also show that HERC2 interacts with, and maintains the levels of, RNF168, another ubiquitin ligase operating downstream of RNF8 (Refs 7, 8). Consequently, knockdown of HERC2 abrogates ubiquitin-dependent retention of repair factors such as 53BP1, RAP80 and BRCA1. Together with the increased radiosensitivity of HERC2-depleted cells, these results uncover a regulatory layer in the orchestration of protein interactions on damaged chromosomes and they underscore the role of ubiquitin-mediated signalling in genome maintenance. PMID- 20023649 TI - GM1 structure determines SV40-induced membrane invagination and infection. AB - Incoming simian virus 40 (SV40) particles enter tight-fitting plasma membrane invaginations after binding to the carbohydrate moiety of GM1 gangliosides in the host cell plasma membrane through pentameric VP1 capsid proteins. This is followed by activation of cellular signalling pathways, endocytic internalization and transport of the virus via the endoplasmic reticulum to the nucleus. Here we show that the association of SV40 (as well as isolated pentameric VP1) with GM1 is itself sufficient to induce dramatic membrane curvature that leads to the formation of deep invaginations and tubules not only in the plasma membrane of cells, but also in giant unilamellar vesicles (GUVs). Unlike native GM1 molecules with long acyl chains, GM1 molecular species with short hydrocarbon chains failed to support such invagination, and endocytosis and infection did not occur. To conceptualize the experimental data, a physical model was derived based on energetic considerations. Taken together, our analysis indicates that SV40, other polyoma viruses and some bacterial toxins (Shiga and cholera) use glycosphingolipids and a common pentameric protein scaffold to induce plasma membrane curvature, thus directly promoting their endocytic uptake into cells. PMID- 20023650 TI - Parallel pathways for vocal learning in basal ganglia of songbirds. AB - The cortical nucleus LMAN (lateral magnocellular nucleus of the anterior nidopallium) provides the output of a basal ganglia pathway that is necessary for vocal learning in juvenile songbirds. The shell subregion of LMAN (LMAN(shell)) gives rise to recurrent loops that may subserve specific learning-related functions. We found that lesions in the LMAN(shell) pathway caused no immediate disruption of vocal behavior, but prevented the development of stable vocal sequences and the ability to imitate vocal sounds. PMID- 20023651 TI - Inactivation of primate superior colliculus impairs covert selection of signals for perceptual judgments. AB - Primates base perceptual judgments on some sensory inputs while ignoring others. The covert selection of sensory information for perception is often thought to be accomplished mostly by the cerebral cortex, whereas the overt orienting toward relevant stimuli involves various additional structures such as the superior colliculus, a subcortical region involved in the control of eye movements. Contrary to this view, we show that the superior colliculus is necessary for determining which stimuli will inform perceptual judgments, even in the absence of orienting movements. Reversible inactivation of the superior colliculus in monkeys performing a motion discrimination task caused profound inattention for stimuli in the affected visual field, but only when distracters containing counterinformative signals appeared in the unaffected field. When distracting stimuli contained no information, discrimination performance was largely unaffected. Thus, the superior colliculus is a bottleneck in the covert selection of signals for perceptual judgments. PMID- 20023652 TI - Activity in the amygdala elicited by unfair divisions predicts social value orientation. AB - 'Social value orientation' characterizes individual differences in anchoring attitudes toward the division of resources. Here, by contrasting people with prosocial and individualistic orientations using functional magnetic resonance imaging, we demonstrate that degree of inequity aversion in prosocials is predictable from amygdala activity and unaffected by cognitive load. This result suggests that automatic emotional processing in the amygdala lies at the core of prosocial value orientation. PMID- 20023653 TI - A robust and high-throughput Cre reporting and characterization system for the whole mouse brain. AB - The Cre/lox system is widely used in mice to achieve cell-type-specific gene expression. However, a strong and universally responding system to express genes under Cre control is still lacking. We have generated a set of Cre reporter mice with strong, ubiquitous expression of fluorescent proteins of different spectra. The robust native fluorescence of these reporters enables direct visualization of fine dendritic structures and axonal projections of the labeled neurons, which is useful in mapping neuronal circuitry, imaging and tracking specific cell populations in vivo. Using these reporters and a high-throughput in situ hybridization platform, we are systematically profiling Cre-directed gene expression throughout the mouse brain in several Cre-driver lines, including new Cre lines targeting different cell types in the cortex. Our expression data are displayed in a public online database to help researchers assess the utility of various Cre-driver lines for cell-type-specific genetic manipulation. PMID- 20023654 TI - The relationship between visual resolution and cone spacing in the human fovea. AB - Visual resolution decreases rapidly outside of the foveal center. The anatomical and physiological basis for this reduction is unclear. We used simultaneous adaptive optics imaging and psychophysical testing to measure cone spacing and resolution across the fovea, and found that resolution was limited by cone spacing only at the foveal center. Immediately outside of the center, resolution was worse than cone spacing predicted and better matched the sampling limit of midget retinal ganglion cells. PMID- 20023655 TI - Control of hippocampal gamma oscillation frequency by tonic inhibition and excitation of interneurons. AB - Gamma-frequency oscillations depend on phasic synaptic GABA(A) receptor (GABA(A)R)-mediated inhibition to synchronize spike timing. The spillover of synaptically released GABA can also activate extrasynaptic GABA(A)Rs, and such tonic inhibition may also contribute to modulating network dynamics. In many neuronal cell types, tonic inhibition is mediated by delta subunit-containing GABA(A)Rs. We found that the frequency of in vitro cholinergically induced gamma oscillations in the mouse hippocampal CA3 region was increased by the activation of NMDA receptors (NMDARs) on interneurons. The NMDAR-dependent increase of gamma oscillation frequency was counteracted by the tonic inhibition of the interneurons mediated by delta subunit-containing GABA(A)Rs. Recordings of synaptic currents during gamma activity revealed that NMDAR-mediated increases in oscillation frequency correlated with a progressive synchronization of phasic excitation and inhibition in the network. Thus, the balance between tonic excitation and tonic inhibition of interneurons may modulate gamma frequency by shaping interneuronal synchronization. PMID- 20023656 TI - State dependence of olfactory perception as a function of taste cortical inactivation. AB - As anyone who has suffered through a head cold knows, food eaten when the olfactory system is impaired tastes 'wrong', an experience that leads many to conclude that taste stimuli are processed normally only when the olfactory system is unimpaired. Evidence that the taste system influences olfactory perception, however, has been vanishingly rare. We found just such an influence; if taste cortex was inactivated when an odor was first presented, later presentations were properly appreciated only if taste cortex was again inactivated. PMID- 20023657 TI - Chimeric mouse tumor models reveal differences in pathway activation between ERBB family- and KRAS-dependent lung adenocarcinomas. AB - To recapitulate the stochastic nature of human cancer development, we have devised a strategy for generating mouse tumor models that involves stepwise genetic manipulation of embryonic stem (ES) cells and chimera generation. Tumors in the chimeric animals develop from engineered cells in the context of normal tissue. Adenocarcinomas arising in an allelic series of lung cancer models containing HER2 (also known as ERBB2), KRAS or EGFR oncogenes exhibit features of advanced malignancies. Treatment of EGFR(L858R) and KRAS(G12V) chimeric models with an EGFR inhibitor resulted in near complete tumor regression and no response to the treatment, respectively, accurately reflecting previous clinical observations. Transcriptome and immunohistochemical analyses reveal that PI3K pathway activation is unique to ERBB family tumors whereas KRAS-driven tumors show activation of the JNK/SAP pathway, suggesting points of therapeutic intervention for this difficult-to-treat tumor category. PMID- 20023658 TI - Genome-wide association study identifies two susceptibility loci for nonsyndromic cleft lip with or without cleft palate. AB - We conducted a genome-wide association study for nonsyndromic cleft lip with or without cleft palate (NSCL/P) in 401 affected individuals and 1,323 controls, with replication in an independent sample of 793 NSCL/P triads. We report two new loci associated with NSCL/P at 17q22 (rs227731, combined P = 1.07 x 10(-8), relative risk in homozygotes = 1.84, 95% CI 1.34-2.53) and 10q25.3 (rs7078160, combined P = 1.92 x 10(-8), relative risk in homozygotes = 2.17, 95% CI 1.32 3.56). PMID- 20023659 TI - Mutations in the formin gene INF2 cause focal segmental glomerulosclerosis. AB - Focal segmental glomerulosclerosis (FSGS) is a pattern of kidney injury observed either as an idiopathic finding or as a consequence of underlying systemic conditions. Several genes have been identified that, when mutated, lead to inherited FSGS and/or the related nephrotic syndrome. These findings have accelerated the understanding of glomerular podocyte function and disease, motivating our search for additional FSGS genes. Using linkage analysis, we identified a locus for autosomal-dominant FSGS susceptibility on a region of chromosome 14q. By sequencing multiple genes in this region, we detected nine independent nonconservative missense mutations in INF2, which encodes a member of the formin family of actin-regulating proteins. These mutations, all within the diaphanous inhibitory domain of INF2, segregate with FSGS in 11 unrelated families and alter highly conserved amino acid residues. The observation that alterations in this podocyte-expressed formin cause FSGS emphasizes the importance of fine regulation of actin polymerization in podocyte function. PMID- 20023660 TI - Wt1 is required for cardiovascular progenitor cell formation through transcriptional control of Snail and E-cadherin. AB - The epicardial epithelial-mesenchymal transition (EMT) is hypothesized to generate cardiovascular progenitor cells that differentiate into various cell types, including coronary smooth muscle and endothelial cells, perivascular and cardiac interstitial fibroblasts and cardiomyocytes. Here we show that an epicardial-specific knockout of the gene encoding Wilms' tumor-1 (Wt1) leads to a reduction in mesenchymal progenitor cells and their derivatives. We show that Wt1 is essential for repression of the epithelial phenotype in epicardial cells and during embryonic stem cell differentiation through direct transcriptional regulation of the genes encoding Snail (Snai1) and E-cadherin (Cdh1), two of the major mediators of EMT. Some mesodermal lineages do not form in Wt1-null embryoid bodies, but this effect is rescued by the expression of Snai1, underscoring the importance of EMT in generating these differentiated cells. These new insights into the molecular mechanisms regulating cardiovascular progenitor cells and EMT will shed light on the pathogenesis of heart diseases and may help the development of cell-based therapies. PMID- 20023661 TI - The activating receptor NKp46 is essential for the development of type 1 diabetes. AB - The mechanism of action of natural killer (NK) cells in type 1 diabetes is still unknown. Here we show that the activating receptor NKp46 recognizes mouse and human ligands on pancreatic beta cells. NK cells appeared in the pancreas when insulitis progressed to type 1 diabetes, and NKp46 engagement by beta cells led to degranulation of NK cells. NKp46-deficient mice had less development of type 1 diabetes induced by injection of a low dose of streptozotocin. Injection of soluble NKp46 proteins into nonobese diabetic mice during the early phase of insulitis and the prediabetic stage prevented the development of type 1 diabetes. Our findings demonstrate that NKp46 is essential for the development of type 1 diabetes and highlight potential new therapeutic modalities for this disease. PMID- 20023662 TI - Thioredoxin-interacting protein links oxidative stress to inflammasome activation. AB - The NLRP3 inflammasome has a major role in regulating innate immunity. Deregulated inflammasome activity is associated with several inflammatory diseases, yet little is known about the signaling pathways that lead to its activation. Here we show that NLRP3 interacted with thioredoxin (TRX)-interacting protein (TXNIP), a protein linked to insulin resistance. Inflammasome activators such as uric acid crystals induced the dissociation of TXNIP from thioredoxin in a reactive oxygen species (ROS)-sensitive manner and allowed it to bind NLRP3. TXNIP deficiency impaired activation of the NLRP3 inflammasome and subsequent secretion of interleukin 1beta (IL-1beta). Akin to Txnip(-/-) mice, Nlrp3(-/-) mice showed improved glucose tolerance and insulin sensitivity. The participation of TXNIP in the NLRP3 inflammasome activation may provide a mechanistic link to the observed involvement of IL-1beta in the pathogenesis of type 2 diabetes. PMID- 20023669 TI - Response to Protocol Review Scenario: Get engaged. PMID- 20023670 TI - A questionable citation and appeal. PMID- 20023671 TI - Response to protocol review scenario: a word from USDA. PMID- 20023672 TI - Response to protocol review scenario: hands off. PMID- 20023663 TI - Shiga toxins--from cell biology to biomedical applications. AB - Shiga toxin-producing Escherichia coli is an emergent pathogen that can induce haemolytic uraemic syndrome. The toxin has received considerable attention not only from microbiologists but also in the field of cell biology, where it has become a powerful tool to study intracellular trafficking. In this Review, we summarize the Shiga toxin family members and their structures, receptors, trafficking pathways and cellular targets. We discuss how Shiga toxin affects cells not only by inhibiting protein biosynthesis but also through the induction of signalling cascades that lead to apoptosis. Finally, we discuss how Shiga toxins might be exploited in cancer therapy and immunotherapy. PMID- 20023673 TI - Response to protocol review scenario: lessons learned. PMID- 20023674 TI - Sudden death in chacma baboons (Papio ursinus). PMID- 20023675 TI - Diagnosis | Acute gastric dilatation. PMID- 20023676 TI - Nasogastric tube placement in the rabbit. AB - If a rabbit is ill or will be undergoing certain types of surgery, it may need to be fed using a nasogastric tube. Nasogastric intubation is easy and is an effective means of delivering nutrition and fluids when other feeding methods are not feasible. This column describes how to place a nasogastric tube in a rabbit. PMID- 20023677 TI - A jugular bleeding technique in rabbits. AB - When studying pharmacokinetics in rabbits, researchers must often take multiple blood samples from conscious rabbits. Researchers usually collect these samples via the auricular vein, typically through a port or an indwelling catheter. The authors have developed an easy and efficient alternative method for obtaining multiple blood samples from conscious rabbits via the external jugular vein. This jugular bleeding technique serves as a refinement to blood sampling methods that require rabbits to undergo surgery (e.g., to insert a port) because it requires no alleviation of pain. During a 2-year period, the authors have taken multiple blood samples from more than 400 rabbits and have seen no adverse events attributed to this procedure. PMID- 20023678 TI - Intraperitoneal catheter placement for pharmacological imaging studies in conscious mice. AB - Imaging studies that use rodents sometimes involve intraperitoneal administration of pharmacological compounds. To facilitate such studies, the authors developed a simple and easily mastered technique for placing an intraperitoneal catheter in a conscious mouse. This technique eliminates the need to remove the animal from the scanner to administer a drug through the intraperitoneal route. PMID- 20023679 TI - The promise of cell-based therapy. PMID- 20023680 TI - Uncontrolled hypertension in children on hemodialysis. PMID- 20023681 TI - Contemporary dialysis: as good as transplantation? PMID- 20023682 TI - Usefulness of RAS inhibition depends on baseline albuminuria. PMID- 20023683 TI - Is angiotensin system blockade indicated in the elderly? PMID- 20023684 TI - Cytomegalovirus prophylaxis: how long is enough? PMID- 20023685 TI - The Framingham risk score is valuable in Europeans. PMID- 20023686 TI - Oral bicarbonate: renoprotective in CKD? PMID- 20023687 TI - Warfarin: safe in dialysis patients with atrial fibrillation? PMID- 20023688 TI - Did 'renalism' skew the results of a negative study? PMID- 20023689 TI - Early intervention in acute kidney injury. AB - The prevalence of acute kidney injury (AKI) in the US is increasing, which can be attributed in part to the aging population and the pandemic of obesity, diabetes and hypertension. Individuals over the age of 70 years constitute 7% of the US population, but account for 36-70% of AKI cases. Presently, major efforts are underway to enable the early detection of AKI by use of various innovative biomarkers, thereby facilitating early intervention. Now is the time, therefore, to consider potential interventions to attenuate or prevent AKI. Several randomized studies have failed to alter the course of AKI, but these studies have been complicated by extrarenal consequences, which could have obscured the potential intrarenal benefit of the agent. Thus, agents that have exclusive effects on renal hemodynamics and tubular function, or the intrarenal infusion of short-acting agents that do not have systemic effects, need to be considered in future studies for the prevention of AKI. PMID- 20023690 TI - Small-cell neuroendocrine carcinoma of the prostate: are heterotransplants a better experimental model? AB - Small-cell neuroendocrine carcinoma of the prostate (SCNCP) is an uncommon type of prostate cancer. However, it is of clinical importance because it is one of the most aggressive tumors of the prostate with a very poor prognosis. There exist few artificially cultured tumor cell lines to study SCNCP. Then, another approach to that study consists in the use of fresh tumor tissue obtained from patients and its heterotransplantation into host mice. The purpose of this review is to integrate data from more than 20 years of heterotransplantation research in the study of small-cell neuroendocrine carcinoma of the prostate (SCNCP). Heterotransplantation has provided data regarding the histopathology, karyotype, DNA content, cell cycle frequency, tumor markers, androgen receptor expression, metastasis and take rate of this prostate disease. When possible, comparisons between original in situ specimens removed from patients and heterotransplanted tissue from host mice have been made. There are advantages, as well as limitations, that have been identified for SCNCP heterotransplants versus xenotransplantation of cultured cells. Overall, heterotransplanted tumors are better than conventional tumor xenografts at retaining tumor morphology, pathology, secretory activity and expression of tumor markers of the patient's original specimen. Furthermore, heterotransplanted tissue preserves the three dimensional tumor architecture of the prostate to maintain critical stromal epithelial cell interactions. PMID- 20023691 TI - Lentivirus-mediated SMO RNA interference inhibits SMO expression and cell proliferation, and affects the cell cycle in LNCaP and PC3 cancer cell lines. AB - Smoothened (SMO) is an important member of the Hedgehog signaling pathway. We constructed a specific recombinant lentiviral vector for RNA interference, targeting the SMO gene (NM_005631) to observe its effect on SMO expression, cell proliferation and the cell cycle in the human androgen-sensitive prostate cancer cell line, LNCaP, and in the androgen-independent prostate cancer cell line, PC3. Four siRNA sequences were designed and inserted into a lentiviral vector pGCSIL GFP to construct four recombinant vectors. The vector with the highest interfering efficiency was co-transfected with packaging vectors (pHelper1.0 and pHelper2.0) in 293T cells to assemble lentivirus particles by liposome for infecting LNCaP and PC3 cell lines, respectively. The expression level of SMO mRNA, tumor cell proliferation and cell cycle were measured by quantitative realtime polymerase chain reaction (qRT-PCR), 3-(4,5)-dimethylthiahiazo (-z-y1) 3,5-di-phenytetrazoliumromide (MTT) assay and flow cytometry, respectively. Sequence results showed that recombinant lentiviral vectors were constructed successfully. pGCSIL-GFP-723 had the highest interfering efficiency, named Lv-SIL SMO723 after co-transfection, with which LNCaP and PC3 cell lines were infected. Compared with the control groups, results showed significantly decreased (P < 0.05) SMO mRNA expressions of LNCaP and PC3, lower mean percentage of S-phase cells and higher mean percentage of G(2)/M phase cells, as well as obviously slow proliferation (P < 0.01) of LNCaP in the infected group. Yet, the proliferation of PC3 was not altered (P > 0.05). In conclusion, the recombinant lentivirus particles were able to suppress SMO expression, regulate the cell cycle in the LNCaP and PC3 cell lines and markedly inhibit proliferation of LNCaP cells but not PC3 cells. PMID- 20023692 TI - Topical delivery of silymarin constituents via the skin route. AB - AIM: Silibinin (SB), silydianin (SD), and silychristin (SC) are components of silymarin. These compounds can be used to protect the skin from oxidative stress induced by ultraviolet (UV) irradiation and treat it. To this end, the absorption of silymarin constituents via the skin was examined in the present report. METHODS: Transport of SB, SD, and SC under the same thermodynamic activity through and into the skin and the effects of pH were studied in vitro using a Franz diffusion assembly. RESULTS: The lipophilicity increased in the order of SC90% of the injected dose accumulates 30 min after injection. Regardless of the presence of the polyanionic inhibitory domain, confocal imaging of live tissues reveals that the majority of the ACPP and CPP remain in punctate organelles, presumably endosomes. Therefore further improvements in the efficiency of delivery to the cytosol and nucleus are necessary. In addition to improved target specificity, a major advantage of ACPPs over CPPs for potential clinical applications is reduced toxicity. Systemically administered r(9) CPP causes acute toxicity in mice at a dose 4-fold lower than the MMP cleavable ACPP, a complication not observed with an uncleavable ACPP presumably because the polycationic charge remains masked systemically. These data suggest that ACPPs have greater potential than CPPs for systemic delivery of imaging and therapeutic agents. PMID- 20023746 TI - Targeting novel integrative nuclear FGFR1 signaling by nanoparticle-mediated gene transfer stimulates neurogenesis in the adult brain. AB - Neurogenesis, the process of differentiation of neuronal stem/progenitor cells (NS/PC) into mature neurons, holds the key to the treatment of various neurodegenerative disorders, which are a major health issue for the world's aging population. We report that targeting the novel integrative nuclear FGF Receptor 1 signaling (INFS) pathway enhances the latent potential of NS/PCs to undergo neuronal differentiation, thus promoting neurogenesis in the adult brain. Employing organically modified silica (ORMOSIL)-DNA nanoplexes to efficiently transfect recombinant nuclear forms of FGFR1 and its FGF-2 ligand into the brain subventricular zone, we find that INFS stimulates the NS/PC to withdraw from the cell cycle, differentiate into doublecortin expressing migratory neuroblasts and neurons that migrate to the olfactory bulb, subcortical brain regions and in the brain cortex. Thus, nanoparticle-mediated non-viral gene transfer may be used to induce selective differentiation of NS/PCs, providing a potentially significant impact on the treatment of a broad range of neurological disorders. PMID- 20023745 TI - In vivo characterization of activatable cell penetrating peptides for targeting protease activity in cancer. AB - Activatable cell penetrating peptides (ACPPs) are novel in vivo targeting agents comprised of a polycationic cell penetrating peptide (CPP) connected via a cleavable linker to a neutralizing polyanion (). Adsorption and uptake into cells are inhibited until the linker is proteolyzed. An ACPP cleavable by matrix metalloproteinase-2 (MMP-2) in vitro was the first one demonstrated to work in a tumor model in vivo, but only HT-1080 xenografts and resected human squamous cell carcinomas were tested. Generality to other cancer types, in vivo selectivity of ACPPs for MMPs, and spatial resolution require further characterization. We now show that ACPPs can target many xenograft tumor models from different cancer sites, as well as a thoroughly studied transgenic model of spontaneous breast cancer (mouse mammary tumor virus promoter driving polyoma middle T antigen, MMTV PyMT). Pharmacological inhibitors and genetic knockouts indicate that current ACPPs are selective for MMP-2 and MMP-9 in the above in vivo models. In accord with the known local distribution of MMP activity, accumulation is strongest at the tumor-stromal interface in primary tumors and associated metastases, indicating better spatial resolution (<50 mum) than other currently available MMP cleavable probes. We also find that background uptake of ACPPs into normal tissues such as cartilage can be decreased by appending inert macromolecules of 30-50 KDa to the polyanionic inhibitory domain. Our results validate an approach that should generally deliver imaging agents and chemotherapeutics to sites of invasion, tumor-promoting inflammation, and metastasis. PMID- 20023747 TI - Multidimensional degradomics identifies systemic autoantigens and intracellular matrix proteins as novel gelatinase B/MMP-9 substrates. AB - The action radius of matrix metalloproteinases or MMPs is not restricted to massive extracellular matrix (ECM) degradation, it extends to the proteolysis of numerous secreted and membrane-bound proteins. Although many instances exist in which cells disintegrate, often in conjunction with induction of MMPs, the intracellular MMP substrate repertoire or degradome remains relatively unexplored. We started an unbiased exploration of the proteolytic modification of intracellular proteins by MMPs, using gelatinase B/MMP-9 as a model enzyme. To this end, multidimensional degradomics technology was developed by the integration of broadly available biotechniques. In this way, 100-200 MMP-9 candidate substrates were isolated, of which 69 were identified. Integration of these results with the known biological functions of the substrates revealed many novel MMP-9 substrates from the intracellular matrix (ICM), such as actin, tubulin, gelsolin, moesin, ezrin, Arp2/3 complex subunits, filamin B and stathmin. About 2/3 of the identified candidates were autoantigens described in multiple autoimmune conditions and in cancer (e.g. annexin I, nucleolin, citrate synthase, HMGB1, alpha-enolase, histidyl-tRNA synthetase, HSP27, HSC70, HSP90, snRNP D3). These findings led to the insight that MMPs and other proteases may have novel (immuno)regulatory properties by the clearance of toxic and immunogenic burdens of abundant ICM proteins released after extensive necrosis. In line with the extracellular processing of organ-specific autoantigens, proteolysis might also assist in the generation of immunodominant 'neo-epitopes' from systemic autoantigens. The study of proteolysis of ICM molecules, autoantigens, alarmins and other crucial intracellular molecules may result in the discovery of novel roles for proteolytic modification. PMID- 20023748 TI - Hematopoietic stem and progenitor cells in adhesive microcavities. AB - The homeostasis of hematopoietic stem and progenitor cells (HSC) in the bone marrow is regulated by a complex interplay of exogenous signals, including extracellular matrix (ECM) molecules, cell-cell contacts, and cytokines. To investigate the influence of spatial restriction and adhesive interactions on HSC fate decisions, we prepared a set of fibronectin-coated micrometer-sized cavities. Analysis of human CD133+ HSCs isolated after culture on these surfaces revealed that proliferation and differentiation is decreased when HSCs are supported by substrates with small microcavities. Single cell analysis of adherent cells also revealed decreased DNA synthesis and higher levels of HSC marker expression inside the smaller cavities. Increasing the cytokine concentration highlighted the tight balance of adhesion related signals and soluble cues acting on HSC fate decisions. Our results suggest that confining human HSCs in ECM-coated microcavities is a possible method to maintain these cells in a quiescent and immature state, an important advantage for several HSC applications. PMID- 20023753 TI - Introducing a new Method section for Integrative Biology. PMID- 20023754 TI - Integration column: biofunctional polymeric nanoparticles for spatio-temporal control of drug delivery and biomedical applications. AB - Polymeric nanoparticle technology has evolved from drug carrier design to advanced multifunctional macromolecular structures. They enable drug delivery and release of a bioactive under spatio-temporal control rather than just passive release by a long-circulating carrier. As such, the carrier is enabling the biomolecule or the bioactive to carry out its designed biological function. Due to their small size nanoparticles may also induce perturbations of biological systems different from any other biomaterials, therefore opening up new biomedical applications as well as raising concerns about adverse effects. PMID- 20023755 TI - Nanomechanical sequencing of collagen: tropocollagen features heterogeneous elastic properties at the nanoscale. AB - Collagen is the most important structural protein in biology and is responsible for the strength and integrity of tissues such as bone, teeth, cartilage and tendon. Here we report a systematic computational sequencing of the effect of amino acid motif variations on the mechanical properties of single tropocollagen molecules, with a particular focus on elastic deformation at varying applied strains. By utilizing a bottom-up computational materiomics approach applied to four model sequence motifs found in human type I collagen, we show that variations in the amino acid motif severely influence the elastic behavior of tropocollagen molecules, leading to softening or stiffening behavior. We also show that interpeptide interactions via H-bonds vary strongly with the type of motif, which implies that it plays a distinct role in the molecule's stability. The most important implication of our results is that deformation in tropocollagen molecules is highly inhomogeneous, since softer regions deform more than stiffer regions, potentially leading to strain and stress concentrations within collagen fibrils. We confirm the hypothesis of inhomogeneous molecular deformation through direct simulation of stretching of a segment of tropocollagen from human type I collagen that features the physiological amino acid sequence. Our results show that the biomechanical properties of tropocollagen must be understood in the context of the specific amino acid sequence as well as the state of deformation, since the elastic properties depend strongly on the amount of deformation applied to a molecule. PMID- 20023756 TI - Directing hepatic differentiation of embryonic stem cells with protein microarray based co-cultures. AB - Embryonic stem cells hold considerable promise in tissue engineering and regenerative medicine as a source of tissue-specific cells. However, realizing this promise requires novel methods for guiding lineage-specific differentiation of stem cells. In this study, we developed a micropatterned co-culture platform for stimulating hepatic differentiation of mouse embryonic stem cells (mESCs). Studies of mESC and hepatic cell adhesion preferences revealed that mESCs required fibronectin for attachment, while hepatic cells (HepG2) preferred collagen (I) substrate and did not adhere to fibronectin. Printing columns of collagen (I) and fibronectin spots (300 microm diameter), followed by sequential seeding of the two cell types, allowed the positioning of clusters of mESCs adjacent to groups of hepatic cells within the same microarray. These micropatterned co-cultures were maintained for up to two weeks in hepatic differentiation media supplemented. To examine the differentiation, mESCs were selectively extracted from the co-culture using laser microdissection and analyzed using real-time reverse transcriptase (RT)-polymerase chain reaction (PCR). These analyses revealed that mESCs co-cultured with HepG2 cells showed a decrease in pluripotency gene expression concomitant with up-regulation of endodermal genes. In addition, the co-culture format induced a significant increase in the expression of liver genes compared to mESCs cultured alone. In conclusion, micropatterned co-cultures of mESCs and hepatic cells showed a significant promise in driving stem cell differentiation towards hepatic phenotype. In the future, this cell culture platform will be further enhanced to enable efficient conversion of mouse and human ESCs to hepatocytes. PMID- 20023757 TI - Adhesion and migration of ovarian cancer cells on crosslinked laminin fibers nanofabricated by multiphoton excited photochemistry. AB - Ovarian cancer is the deadliest gynecological cancer, which may arise in part due to the concurrent invasion and metastasis of high grade tumors. It is thus crucial to gain insight into the adhesion and migration mechanisms in vivo, as this may ultimately lead to new treatment/detection options. To explore this possibility, we have used multiphoton excited photochemistry (MPE) to synthesize models of the ovarian basal lamina consisting of crosslinked laminin nanofibers to quantify the adhesion/migration dynamics. The nanostructured laminin patterns permit the systematic comparison of total migration, directed migration, adhesion, and morphology of "normal" immortalized human ovarian epithelial cells (IOSE) and three lines of varying metastatic potential (OVCA433, SKOV-3.ip1, and HEY-1 cells). We find that the migration of all the cell lines is directed by the crosslinked fibers, and that the contact guidance enhances the total migration rates relative to monolayers. These rates increase with increasing metastatic potential, and the more invasive cells are less rigid and more weakly adhered to the nanofibers. The extent of directed migration also depends on the cell polarity and focal adhesion expression. For the invasive cells, these findings are similar to the integrin-independent ameboid-like migration seen for polar cells in collagen gels. Collectively, the results suggest that contact mediated migration as well as decreased adhesion may be operative in metastasis of ovarian cancer in vivo. PMID- 20023758 TI - Dielectric characterization of complete mononuclear and polymorphonuclear blood cell subpopulations for label-free discrimination. AB - Dielectric spectroscopy is a powerful technique for the elucidation of a number of important cell biophysical properties, and it can provide information about cell morphology, physiological state, viability and identity. A high-impact application for dielectric cell analysis would be microfluidic flow-through impedance sensing to perform what is perhaps the most routinely ordered medical diagnostic assay, a complete blood count with white blood cell differential enumeration. To assess the biophysical feasibility of such an analysis, we obtained reference dielectric measurements of the complete complement of purified leukocyte subpopulations using the dielectrophoretic crossover frequency method. The sensitivity of this method can detect subtle changes in cell morphology and physiology, so we developed a leukocyte isolation protocol based on a suite of negative selection techniques to yield cell subpopulations that were minimally processed and in an as near native state as possible. This is the first reported study of the dielectric properties of all the major leukocyte subpopulations that includes separate analysis of the polymorphonuclear neutrophil, basophil and eosinophil cell types. We show that T-lymphocytes, B-lymphocytes, monocytes and granulocytes possess distinct membrane dielectric properties and that the morphologically similar granulocyte subpopulations can be identified via their dielectric and size properties. Finally, we discuss the application of our findings to label-free systems for the analysis of leukocytes. PMID- 20023764 TI - Application of aptamers and autofluorescent proteins for RNA visualization. AB - The repertoire of RNAs transcribed and processed within living cells is of extraordinary complexity. With new types of RNA being identified, the need for tools to investigate the spatio-temporal aspects of processing and trafficking of these molecules has become more evident. To visualize RNA in living cells, autofluorescent proteins (AFPs) appear as a promising alternative to synthetic fluorescent compound based labels. While current fluorescent protein-based RNA labelings have provided many new insights into the biology of RNA regulation, further improvements and adaptations are desirable to make AFP labels as valuable in the RNA world as they have proven to be for protein tagging. This article reviews the achievements and existing challenges in engineering AFPs as efficient RNA tags for high resolution fluorescence microscopy in living cells. PMID- 20023766 TI - Investigating the role of the extracellular environment in modulating hepatic stellate cell biology with arrayed combinatorial microenvironments. AB - Hepatic stellate cells (HSCs) are a major cell type of the liver that are involved in liver homeostasis. Upon liver damage, HSCs exit their normally quiescent state and become activated, leading to an increase of their proliferation, production of abnormal extracellular matrix proteins (ECMPs) and inflammatory mediators, and eventually liver fibrosis and cirrhosis. Current in vitro approaches to identify components that influence HSC biology typically investigate one factor at a time and generally ignore the complex crosstalk among the myriad of components that comprise the microenvironments of quiescent or activated HSCs. Here we describe a high throughput screening (HTS) approach to identify factors that affect HSC biology. Specifically, we integrated the use of ECMPs and signaling molecules into a combinatorial cellular microarray technology platform, thereby creating comprehensive "microenvironments". Using this technology, we performed real-time simultaneous screening of the effects of hundreds of unique microenvironments composed of ECMPs and signaling molecules on HSC proliferation and activation. From these screens, we identified combinations of microenvironment components that differentially modulate the HSC phenotype. Furthermore, analysis of HSC responses revealed that the influences of Wnt signaling molecules on HSC fate are dependent on the ECMP composition in which they are presented. Collectively, our results demonstrate the utility of high content, array-based screens to provide a better understanding of HSC biology. Our results indicate that array-based screens may provide an efficient means for identifying candidate signaling pathways to be targeted for anti-fibrotic therapies. PMID- 20023765 TI - Spontaneous migration of cancer cells under conditions of mechanical confinement. AB - When cancer cells spread away from the primary tumor, they often follow the trajectories of lymphatic vessels, nerves, white matter tracts, or other heterogeneous structures in tissues. To better understand this form of guided cell migration we designed a series of microfluidic devices that mechanically constrain migrating cancer cells inside microchannels with cross-section comparable to cell size. We observed unexpectedly fast and persistent movement in one direction for several hours of cancer cells of different types. The persistent motility occurs spontaneously, in the absence of external gradients, suggesting the presence of intrinsic mechanisms driving cancer cell motility that are induced in conditions of mechanical confinement. To probe the mechanisms responsible for this behavior, we exposed cancer cells inside channels to drugs targeting the microtubules, and measured a significant reduction in the average migration speed. Surprisingly, a small number of cells appeared not to be affected by the treatment and displayed fast and persistent migration, comparable to the untreated cells. The new matrix-free, 3D-confined motility assay replicates critical interactions that cancer cells would normally have inside tissues, is compatible with high-content, high-throughput analysis of cellular motility at single cell level, and could provide useful insights into the biology of cancer cell migratory phenotype. PMID- 20023767 TI - Size selective behavior of mesenchymal stem cells on ZrO(2) and TiO(2) nanotube arrays. AB - This work reports on the behavior of mesenchymal stem cells on anodic ZrO(2) nanotube layers grown by a self-ordering process on zirconium with defined diameters between 10 and 50 nm. It is demonstrated that mesenchymal stem cells show a size-specific reaction to these nanoscale patterned surfaces. We compare the behavior on these ZrO(2) nanotubes to findings on TiO(2) nanotubes of different diameters. For both nanotube materials, TiO(2) and ZrO(2), cell adhesion and spreading are enhanced for nanotube diameters of approximately 15-30 nm, while a strong decay in cell activity is observed for diameters >50 nm. Focal complex formation on adherent cells is selectively modulated by the specific nanoscale. Moreover, even if the surface chemistry of the nanotubes is completely modified with a dense AuPd coating onto the formed nanotube layers, or the length of the nanotubes is varied, the observed nano size effects still prevail. This demonstrates how strong the pure geometric diameter dependence in the range between 15 and 100 nm dominates over other possible effects on cell activity. PMID- 20023768 TI - Impaired interaction between skeletal ryanodine receptors in malignant hyperthermia. AB - Functional coupling between clustered membrane receptors has been identified as a novel mechanism to improve signaling performance in a number of physiological processes. The potential role of defective inter-receptor coupling in the pathogenesis of disease, however, has not previously been explored. Ryanodine receptors (RyRs), the primary calcium release channel of muscle, usually form ordered two-dimensional arrays in the sarcoplasmic reticulum membranes. Mutations in RyRs are known to cause a number of severe diseases both in skeletal muscle and in heart. Here we present a model demonstrating how impaired functional coupling between neighboring mutant RyR1(R615C) channels may contribute to the pharmacogenetic skeletal muscle sensitivity, malignant hyperthermia (MH). We find that purified RyR1(R615C) from MH susceptible porcine skeletal muscle shows significantly reduced oligomerization when compared to RyR1(WT), indicating a potential loss of intrinsic intermolecular control. The MH-triggering volatile anesthetic, halothane, activates RyR1(R615C) and RyR1(WT) to a similar extent, using [(3)H]ryanodine binding as a measure of activation. Modeling RyR1 array function with parameters modified to simulate the loss of functional inter-RyR coupling recapitulates the MH molecular phenotype-RyR1 channels leaky to Ca(2+) at rest and long open-times following exposure to halothane. Our work suggests that a defect in inter-RyR1 coupling is a novel direction for research into the pathogenesis of MH. PMID- 20023769 TI - Single-cell transcript analysis of human embryonic stem cells. AB - We demonstrate the qualitative and quantitative power of single-cell transcript analysis to characterize transcriptome dynamics in human embryonic stem cells (hESC's). Single-cell analysis can systematically determine unique cellular profiles for use in cell sorting and identification, show the potential to augment standing models of cellular differentiation, and elucidate the behavior of stem cells exiting pluripotency. Using single-cell analysis of H9 hESC's differentiating under three culture conditions, we revealed transient expression of mesendodermal markers in all three protocols, followed by increasingly stable expression of embryonic endoderm and extra-embryonic endoderm markers. Our single cell profiles reveal mixed populations of cell types, with both transcriptional and temporal heterogeneity marking differentiation under all conditions. Interestingly, we also observe extensive and prolonged co-expression of markers regulating both pluripotency and lineage differentiation in all culture conditions, and we find that pluripotency marker transcripts remain detectable in the majority of cells for many days. Finally, we show that cells derived from undifferentiated hESC colonies display consistent gene expression profiles characterized by three cohorts of transcripts: uniform, absent and sporadically detected messages, and that a striking correlation exists between genes' membership in these cohorts and their hESC promoter chromatin state, with bivalent promoters dominating the sporadic transcripts. PMID- 20023774 TI - Microfluidic sample preparation: cell lysis and nucleic acid purification. AB - Due to the lack of development in the area of sample preparation, few complete lab-on-a-chip systems have appeared in recent years that can deal with raw samples. Cell lysis and nucleic acid extraction systems are sufficiently complex even before adding the complexity of an analysis system. In this review, a variety of microfluidic sample preparation methods are discussed and evaluated. Microsystems for cell lysis are discussed by grouping them into categories based on their lysis mechanisms: mechanical, chemical, thermal or electrical. We classify the nucleic acid purification techniques according to the mechanism that links nucleic acids to substrates: silica-based surface affinity, electrostatic interaction, nanoporous membrane filtration, and functionalized microparticles. The techniques for microfluidic cell lysis and nucleic acid purification are compared based on the ease of microfabrication and integration, and sample flexibility. These assessments can help us determine the appropriate sample preparation technique for generating a true lab-on-a-chip. PMID- 20023775 TI - Parylene peel-off arrays to probe the role of cell-cell interactions in tumour angiogenesis. AB - Microenvironmental conditions impact tumour angiogenesis, but the role of cell cell interactions in modulating the angiogenic capability of tumour cells is not well understood. We have microfabricated a peel-off cell-culture array (PeelArray) chip to spatiotemporally control interactions between tumour cells in a large array format and to analyse angiogenic factor secretion in response to these conditions. The PeelArray chip consists of a polyethylene glycol (PEG) treated glass coverslip coated with a parylene-C template that can be easily peeled off to selectively micropattern biomolecules and cells. We have designed the PeelArray chip to reproducibly deposit large uniform arrays of isolated single cells or isolated cell clusters on fibronectin features of defined surface areas. We have utilised this microfabricated culture system to study the secretion of angiogenic factors by tumour cells, in the presence or absence of cell-cell contact as controlled by micropatterning. Our results indicate that cell-cell interactions play a synergistic role in regulating the expression of angiogenic factors (i.e., vascular endothelial growth factor [VEGF] and interleukin-8 [IL-8]) in various cancer cell lines, independent of other more complex microenvironmental cues (e.g. hypoxia). Our PeelArray chip is a simple and adaptable micropatterning method that enables quantitative profiling of protein secretions and hence, a better understanding of the mechanisms by which cell-cell interactions regulate tumour cell behaviour and angiogenesis. PMID- 20023776 TI - Cell-cell communication between adipocytes and pancreatic beta-cells in acoustically levitated droplets. AB - Dysfunctional adipocytes and insulin-producing pancreatic beta-cells are hallmarks of human Type 2 diabetes and play important roles in the onset and progression of the disease. However, the precise mechanisms involved are complex and only partially understood. Here we present a new and unique method to perform single-cell and cell-cell communication studies in Type 2 diabetes-related research. The airborne analytical system offers "contactless" sample handling in the sub-microlitre volume range and is here equipped with fluorescence imaging detection. The system utilizes acoustically levitated droplets as "wall-less" test tubes and in-house constructed piezoelectric flow-through picolitre droplet dispensers for precise reagent supply. Hormone-mediated regulation of adipocyte lipolysis and communication between adipocytes and beta-cells can be studied at the few-cell level. Thus, lipolysis could be detected in single adipocytes, whether it was induced by isoprenaline or inhibited by insulin. Furthermore, the airborne system allowed the comparison of lipolysis in adipocytes of different sizes: a large adipocyte responded more slowly than a small cell. Furthermore, stimulation of insulin secretion by high glucose or acetylcholine administration to a levitated drop containing insulin-producing beta-cells resulted in inhibition of isoprenaline-induced lipolysis in adipocytes present in the same drop. The results show the applicability of the airborne analytical system for single cell analysis and for cell-cell communication studies as well as the potential for future analysis directly from human cells obtained from clinical biopsies. PMID- 20023777 TI - Millisecond timescale slimfield imaging and automated quantification of single fluorescent protein molecules for use in probing complex biological processes. AB - Fluorescence microscopy offers a minimally perturbative approach to probe biology in vivo. However, available techniques are limited both in sensitivity and temporal resolution for commonly used fluorescent proteins. Here we present a new imaging system with a diagnostic toolkit that caters for the detection and quantification of fluorescent proteins for use in fast functional imaging at the single-molecule level. It utilizes customized microscopy with a mode of illumination we call "slimfield" suitable for rapid (approximately millisecond) temporal resolution on a range of common fluorescent proteins. Slimfield is cheap and simple, allowing excitation intensities approximately 100 times greater than those of widefield imaging, permitting single-molecule detection at high speed. We demonstrate its application on several purified fluorescent proteins in standard use as genetically-encoded reporter molecules. Controlled in vitro experiments indicate single protein molecules over a field of view of approximately 30 microm(2) area, large enough to encapsulate complete prokaryotic and small eukaryotic cells. Using a novel diagnostic toolkit we demonstrate automated detection and quantification of single molecules with maximum imaging rates for a 128 x 128 pixel array of approximately 500 frames per second with a localization precision for these photophysically poor fluorophores to within 50 nm. We report for the first time the imaging of the dim enhanced cyan fluorescent protein (ECFP) and CyPet at the single-molecule level. Applying modifications, we performed simultaneous dual-colour slimfield imaging for use in co-localization and FRET. We present preliminary in vivo imaging on bacterial cells and demonstrate approximately millisecond timescale functional imaging at the single molecule level with negligible photodamage. PMID- 20023786 TI - Integrating biology and technology. PMID- 20023788 TI - Integration column: Microfluidic high-throughput screening. AB - Biology has always been a heavily technology limited field. Burgeoning fields such as systems biology require the development and implementation of new technologies, enabling high-throughput and high-fidelity measurements of large systems. Microfluidics promises to fulfil many of the requirements put forth. Here I will discuss the various approaches employed to date for performing high throughput screening experiments on-chip, encompassing biochemical, biophysical, and cell-based assays. PMID- 20023789 TI - Manipulation of biological samples using micro and nano techniques. AB - The constant interest in handling, integrating and understanding biological systems of interest for the biomedical field, the pharmaceutical industry and the biomaterial researchers demand the use of techniques that allow the manipulation of biological samples causing minimal or no damage to their natural structure. Thanks to the advances in micro- and nanofabrication during the last decades several manipulation techniques offer us the possibility to image, characterize and manipulate biological material in a controlled way. Using these techniques the integration of biomaterials with remarkable properties with physical transducers has been possible, giving rise to new and highly sensitive biosensing devices. This article reviews the different techniques available to manipulate and integrate biological materials in a controlled manner either by sliding them along a surface (2-D manipulation), by grapping them and moving them to a new position (3-D manipulation), or by manipulating and relocating them applying external forces. The advantages and drawbacks are mentioned together with examples that reflect the state of the art of manipulation techniques for biological samples (171 references). PMID- 20023790 TI - Raman tweezers and their application to the study of singly trapped eukaryotic cells. AB - In this review the recent emergence of Raman tweezers as an analytical technique for single eukaryotic cell analysis is described. The Raman tweezer technique combines Raman spectroscopy as a diagnostic tool with optical tweezers by which means single cells can be trapped and manipulated in a laser beam using a high numerical aperture imaging microscope. Necessary instrumental requirements to facilitate Raman tweezer experiments are discussed together with practical considerations such as the potential for photodamage of cells subjected to trapping and Raman excitation. Specific applications of Raman tweezers to the analysis of cancer cells, erythrocytes and lymphocytes, micro-organisms and sub cellular components e.g.chromosomes and mitochondria are then discussed followed by a summary of the future potential of the technique for single cell analysis. PMID- 20023791 TI - Advances and perspectives in aptamer arrays. AB - Aptamers are oligonucleotides (typically 10-60 bases in length) capable of binding target ligands with affinities similar to antibodies. The generation of high density multiplexed aptamer arrays for molecular diagnostics was first proposed nearly ten years ago for the quantification of the thousands of proteins within biological samples, including blood and urine. The tagless aptameric detection of small molecular compounds extends the application of such arrays to bioanalyses at the metabolite level. We present here a minireview on some existing technologies and highlight recent innovations that are being applied to this field, which may facilitate the vision of highly multi-parallelized arrays for the quantitative analysis of biological systems. PMID- 20023792 TI - Perturbation of single hematopoietic stem cell fates in artificial niches. AB - Hematopoietic stem cells (HSCs) are capable of extensive self-renewal in vivo and are successfully employed clinically to treat hematopoietic malignancies, yet are in limited supply as in culture this self-renewal capacity is lost. Using an approach at the interface of stem cell biology and bioengineering, here we describe a novel platform of hydrogel microwell arrays for assessing the effects of either secreted or tethered proteins characteristic of the in vivo microenvironment, or niche, on HSC fate in vitro. Time-lapse microscopic analyses of single cells were crucial to overcoming inevitable heterogeneity of FACS enriched HSCs. A reduction in proliferation kinetics or an increase in asynchronous division of single HSCs in microwells in response to specific proteins (Wnt3a and N-Cadherin) correlated well with subsequent serial long-term blood reconstitution in mice in vivo. Single cells that divided once in the presence of a given protein were capable of in vivo reconstitution, providing evidence of self-renewal divisions of HSCs in vitro. These results validate the hydrogel microwell platform as a broadly applicable paradigm for dissecting the regulatory role of specific signals within a complex stem cell niche. PMID- 20023793 TI - Human mammary progenitor cell fate decisions are products of interactions with combinatorial microenvironments. AB - In adult tissues, multi-potent progenitor cells are some of the most primitive members of the developmental hierarchies that maintain homeostasis. That progenitors and their more mature progeny share identical genomes, suggests that fate decisions are directed by interactions with extrinsic soluble factors, ECM, and other cells, as well as physical properties of the ECM. To understand regulation of fate decisions, therefore, would require a means of understanding carefully choreographed combinatorial interactions. Here we used microenvironment protein microarrays to functionally identify combinations of cell-extrinsic mammary gland proteins and ECM molecules that imposed specific cell fates on bipotent human mammary progenitor cells. Micropatterned cell culture surfaces were fabricated to distinguish between the instructive effects of cell-cell versus cell-ECM interactions, as well as constellations of signaling molecules; and these were used in conjunction with physiologically relevant 3 dimensional human breast cultures. Both immortalized and primary human breast progenitors were analyzed. We report on the functional ability of those proteins of the mammary gland that maintain quiescence, maintain the progenitor state, and guide progenitor differentiation towards myoepithelial and luminal lineages. PMID- 20023794 TI - Mapping mammary gland architecture using multi-scale in situ analysis. AB - We have built a novel computational microscopy platform that integrates image acquisition, storage, processing and analysis to study cell populations in situ. This platform allows high-content studies where multiple features are measured and linked at multiple scales. We used this approach to study the cellular composition and architecture of the mouse mammary gland by quantitatively tracking the distribution and type, position, proliferative state, and hormone receptor status of epithelial cells that incorporated bromodeoxyuridine while undergoing DNA synthesis during puberty and retained this label in the adult gland as a function of tissue structure. Immunofluorescence was used to identify label-retaining cells, as well as epithelial cells expressing the proteins progesterone receptor and P63. Only 3.6% of luminal cells were label-retaining cells, the majority of which did not express the progesterone receptor. Multi scale in situ analysis revealed that luminal label-retaining cells have a distinct nuclear morphology, are enriched 3.4-fold in large ducts, and are distributed asymmetrically across the tissue. We postulated that LRC enriched in the ventral mammary gland represent progenitor cells. Epithelial cells isolated from the ventral versus the dorsal portion of the gland were enriched for the putative stem cell markers CD24 and CD49f as measured by fluorescence activated cell sorting. Thus, quantitative analysis of the cellular composition of the mammary epithelium across spatial scales identified a previously unrecognized architecture in which the ventral-most, large ducts contain a reservoir of undifferentiated, putative stem cells. PMID- 20023795 TI - Real-time detection of the early event of cytotoxicity of herbal ingredients on single leukemia cells studied in a microfluidic biochip. AB - A microfluidic approach has been developed for the real-time detection of drug effects, based on the quantitative measurement of calibrated cytosolic calcium ([Ca(2+)](i)) on single cancer cells. This microfluidic method is rapid by detecting the early event of cytotoxicity of drug candidates on cancer cells, without waiting for a couple of days needed for cell seeding and drug treatment by conventional assays. The miniaturized biochip consists of a V-shaped structure for the single-cell selection and retention. Various test reagents such as the chemotherapy drug (daunorubicin), an ionophore (ionomycin), and herbal ingredients from licorice (isoliquiritigenin or IQ) were investigated for their abilities to stimulate sustained cellular [Ca(2+)](i) elevations. The microfluidic results obtained in hours have been confirmed by conventional cytotoxicity assays which take days to complete. Moreover, any color or chemical interference problems found in the conventional assays of herbal compounds could be resolved. Using the microfluidic approach, IQ (50 microM) has been found to cause a sustained [Ca(2+)](i) elevation and cytotoxic effects on leukemia cells. The microfluidic single-cell analysis not only reduces reagent cost, and demands less cells, but also reveals some phenomena due to cellular heterogeneity that cannot be observed in bulk analysis. PMID- 20023796 TI - Genome-wide transcriptome analysis of 150 cell samples. AB - A major challenge in molecular biology is interrogating the human transcriptome on a genome wide scale when only a limited amount of biological sample is available for analysis. Current methodologies using microarray technologies for simultaneously monitoring mRNA transcription levels require nanogram amounts of total RNA. To overcome the sample size limitation of current technologies, we have developed a method to probe the global gene expression in biological samples as small as 150 cells, or the equivalent of approximately 300 pg total RNA. The new method employs microfluidic devices for the purification of total RNA from mammalian cells and ultra-sensitive whole transcriptome amplification techniques. We verified that the RNA integrity is preserved through the isolation process, accomplished highly reproducible whole transcriptome analysis, and established high correlation between repeated isolations of 150 cells and the same cell culture sample. We validated the technology by demonstrating that the combined microfluidic and amplification protocol is capable of identifying biological pathways perturbed by stimulation, which are consistent with the information recognized in bulk-isolated samples. PMID- 20023797 TI - Influence of applied currents on the viability of cells close to microelectrodes. AB - Electrodes have become more and more popular in biomedical and bioengineering applications, where they are used mostly to apply/measure potentials or currents to/from biological systems. Under such conditions, electrochemical reactions commonly occur at the electrode surface. With the aim to better describe these processes we applied constant currents using transparent indium tin oxide microelectrodes to induce a local change in pH, associated with electrolysis. The pH change was monitored optically within the first lateral 170 microm vicinity using microscopy and a pH sensitive fluorescent dye combination as indicator. The data were then fitted with a simple diffusion model. The effect of such an induced pH change was also assessed by measuring the desorption of a cationic polyelectrolyte (poly(l-lysine)-grafted-poly(ethylene glycol)) as a function of the local pH. Since this polymer interacts electrostatically with surfaces in a pH dependent manner, we could show a strong effect in unbuffered solutions while buffered solutions restricted the electrochemically induced pH change below the optical resolution of the microscope. The effect of applied current on the behavior of cells was also studied on myoblasts cultured directly on the microelectrodes. We have found that current densities larger than 0.57 A m(-2) induced cell death within 2 min of exposure. Based on our model we could attribute this to the change in local pH although the effect of other electrochemically created reactive molecules could not be excluded. PMID- 20023798 TI - Analysis of aptamer sequence activity relationships. AB - DNA sequences that can bind selectively and specifically to target molecules are known as aptamers. Normally such binding analyses are performed using soluble aptamers. However, there is much to be gained by using an on-chip or microarray format, where a large number of aptameric DNA sequences can be interrogated simultaneously. To calibrate the system, known thrombin binding aptamers (TBAs) have been mutated systematically, producing large populations that allow exploration of key structural aspects of the overall binding motif. The ability to discriminate between background noise and low affinity binding aptamers can be problematic on arrays, and we use the mutated sequences to establish appropriate experimental conditions and their limitations for two commonly used fluorescence based detection methods. Having optimized experimental conditions, high-density oligonucleotide microarrays were used to explore the entire loop-sequence functionality relationship creating a detailed model based on over 40 000 analyses, describing key features for quadruplex-forming sequences. PMID- 20023799 TI - The development of integrative biology: bridging the gap--a view from the Scientific Editors. [An interview with David Beebe and Mary Helen Barcellos-Hoff by Kathleen Too]. PMID- 20023800 TI - Models of protein linear molecular motors for dynamic nanodevices. AB - Protein molecular motors are natural nano-machines that convert the chemical energy from the hydrolysis of adenosine triphosphate into mechanical work. These efficient machines are central to many biological processes, including cellular motion, muscle contraction and cell division. The remarkable energetic efficiency of the protein molecular motors coupled with their nano-scale has prompted an increasing number of studies focusing on their integration in hybrid micro- and nanodevices, in particular using linear molecular motors. The translation of these tentative devices into technologically and economically feasible ones requires an engineering, design-orientated approach based on a structured formalism, preferably mathematical. This contribution reviews the present state of the art in the modelling of protein linear molecular motors, as relevant to the future design-orientated development of hybrid dynamic nanodevices. PMID- 20023801 TI - The evolution of chemotaxis assays from static models to physiologically relevant platforms. AB - The role of chemotactic gradients in the immunological response is an area which elicits a lot of attention due to its impact on the outcome of the inflammatory process. Consequently there are numerous standard in vitro designs which attempt to mimic chemotactic gradients, albeit in static conditions, and with no control over the concentration of the chemokine gradient. In recent times the design of the standard chemotaxis assay has incorporated modern microfluidic platforms, computer controlled flow devices and cell tracking software. Assays under fluid flow which use biochips have provided data which highlight the importance of shear stress on cell attachment and migration towards a chemokine gradient. However, the in vivo environment is far more complex in comparison to conventional cell assay chambers. The designs of biochips are therefore in constant flux as advances in technology permit ever greater imitations of in vivo conditions. Researchers are focused on designing a generation of new biochips and enhancing the physiological relevance of the current assays. The challenge is to combine a shear flow with a 3D scaffold containing the endothelial layer and permitting a natural diffusion of chemokines through a tissue-like basal matrix. Here we review the latest range of chemotaxis assays and assess the innovative features of their designs which enable them to better imitate the in vivo environment. We also present some alternative designs that were initially employed in tissue engineering which could potentially be used in the establishment of novel chemotaxis assays. PMID- 20023802 TI - From the cellular perspective: exploring differences in the cellular baseline in macroscale and microfluidic cultures. AB - Microfluidic devices for cell culture based assays provide new types of engineered microenvironments and new methods for controlling and quantifying cellular responses to these microenvironments. However, without an understanding of the effects of the microenvironments present in microdevices from a cellular perspective, it will be challenging to integrate work done in microdevices with biological data obtained via traditional methods. With the adaptation and validation of In Cell Westerns (ICWs) and in situ analysis techniques to microfluidic devices, we can begin to look at a variety of cellular responses to microcultures. Here we observe several differences in proliferation, glucose metabolism, signaling pathway activation and protein expression levels between cells cultured in traditional macroscale cultures and in microfluidic cultures. The issues of glucose starvation, growth factor restriction, volume density and effects of interactions with poly(dimethylsiloxane) (PDMS) were examined to determine the relative importance of each to cell behavior. Changes in glucose metabolism, insensitivity to volume density or media supplementation, and finally reduced proliferation as the exposure to PDMS increased, suggests that perhaps interactions between media/cells and this commonly employed polymer may be significant for some cell based assays. The differences between cells in macroscale and microfluidic cultures suggest that the cellular baseline may be substantially altered in microcultures due to both inherent differences in scale as well as material differences. The observations highlight the need to biologically validate micofluidic devices for cell based assays in order to accurately interpret the data obtained with them in the context of traditional macroculture data. Additional areas of study that will further characterize and validate microscale culture are discussed. PMID- 20023803 TI - Surface topography induces 3D self-orientation of cells and extracellular matrix resulting in improved tissue function. AB - The organization of cells and extracellular matrix (ECM) in native tissues plays a crucial role in their functionality. However, in tissue engineering, cells and ECM are randomly distributed within a scaffold. Thus, the production of engineered-tissue with complex 3D organization remains a challenge. In the present study, we used contact guidance to control the interactions between the material topography, the cells and the ECM for three different tissues, namely vascular media, corneal stroma and dermal tissue. Using a specific surface topography on an elastomeric material, we observed the orientation of a first cell layer along the patterns in the material. Orientation of the first cell layer translates into a physical cue that induces the second cell layer to follow a physiologically consistent orientation mimicking the structure of the native tissue. Furthermore, secreted ECM followed cell orientation in every layer, resulting in an oriented self-assembled tissue sheet. These self-assembled tissue sheets were then used to create 3 different structured engineered-tissue: cornea, vascular media and dermis. We showed that functionality of such structured engineered-tissue was increased when compared to the same non-structured tissue. Dermal tissues were used as a negative control in response to surface topography since native dermal fibroblasts are not preferentially oriented in vivo. Non structured surfaces were also used to produce randomly oriented tissue sheets to evaluate the impact of tissue orientation on functional output. This novel approach for the production of more complex 3D tissues would be useful for clinical purposes and for in vitro physiological tissue model to better understand long standing questions in biology. PMID- 20023804 TI - Manipulation and charge determination of proteins in photopatterned solid supported bilayers. AB - This work demonstrates the use of deep UV micropatterned chlorotrimethylsilane (TMS) monolayers to support lipid membranes on SiO(2) surfaces. After immersing such a patterned surface into a solution containing small unilamellar vesicles of egg PC, supported bilayer lipid membranes were formed on the hydrophilic, photolyzed regions and lipid monolayer over the hydrophobic, non-photolyzed regions. A barrier between the lipid monolayer and bilayer regions served to stop charged lipids migrating between the two. This allows the system to be used to separate charged lipids or proteins by electrophoresis. Either oppositely charged fluorescence labeled lipids [Texas Red DHPE (negative charge) and D291 (positive charge)] or lipids with different charge numbers [Texas Red DHPE (one negative charge) and NBD PS (two negative charges)] can be separated. We have also studied the migration of streptavidin attached to a biotinylated lipid. Negatively charged streptavidin responds to the applied electric field by moving in the direction of electroosmotic flow, i.e. towards the negative electrode. However the direction of streptavidin movement can be controlled by altering the difference in zeta potential between that of the streptavidin (zeta(1)) and the lipid membrane (zeta(2)). If zeta(1) > zeta(2), streptavidin moves to the negative electrode, while if zeta(1) < zeta(2), streptavidin moves to the positive electrode. This balance was manipulated by adding positively charged lipid DOTAP to the membrane. After measuring the average drift velocity of streptavidin as a function of DOTAP concentration, the point where zeta(1) approximately zeta(2) was found. At this point zeta(1) was calculated to be -9.8 mV which is in good agreement with the value of -13 mV from force measurements and corresponds to a charge of -2e per streptavidin, thus demonstrating the applicability of this method for determining protein charge. PMID- 20023805 TI - Mechanical and spatial determinants of cytoskeletal geodesic dome formation in cardiac fibroblasts. AB - This study tests the hypothesis that the cell cytoskeletal (CSK) network can rearrange from geodesic dome type structures to stress fibers in response to microenvironmental cues. The CSK geodesic domes are highly organized actin microarchitectures within the cell, consisting of ordered polygonal elements. We studied primary neonatal rat cardiac fibroblasts. The cues used to trigger the interconversion between the two CSK architectures (geodesic domes and stress fibers) included factors affecting spatial order and the degree of CSK tension in the cells. Microfabricated three-dimensional substrates with micrometre sized grooves and peaks were used to alter the spatial order of cell growth in culture. CSK tension was modified by 2,3-butanedione 2-monoxime (BDM), cytochalasin D and the hyphae of Candida albicans. CSK geodesic domes occurred spontaneously in about 20% of the neonatal rat cardiac fibroblasts used in this study. Microfabricated structured surfaces produced anisotropy in the cell CSK and effectively converted geodesic domes into stress fibers in a dose-dependent manner (dependence on the period of the features). Affectors of actin structure, inhibitors of CSK tension and cell motility, e.g. BDM, cytochalasin D and the hyphae of C. albicans, suppressed or eliminated the geodesic domes. Our data suggest that the geodesic domes, similar to actin stress fibers, require maintenance of CSK integrity and tension. However, microenvironments that promote structural anisotropy in tensed cells cause the transformation of the geodesic domes into stress fibers, consistent with topographic cell guidance and some previous CSK model predictions. PMID- 20023806 TI - Empirically corrected DFT and semi-empirical methods for non-bonding interactions. AB - Computational modeling of systems governed by non-bonded interactions, especially van der Waals (dispersion) interactions, is currently a difficult task, since many conventional quantum mechanical techniques neglect such interactions. For example, the popular semi-empirical and Hartree-Fock methods, as well as most DFT methods all neglect long-range dispersion. In attempt to model dispersion interactions at reduced computational expense, one approach is to add an empirical potential to one of the quantum mechanical techniques. This approach has been successfully used to model a large variety of systems that involve, or are governed by, dispersion interactions. The accuracy of empirically-corrected density functional theory (DFT-D) and empirically-corrected semiempirical (SE-D) methods are reviewed here. The analysis considers both the ability to reproduce benchmark energies and geometries. PMID- 20023807 TI - Similarity between the kinetic parameters of the buffer-mediated proton exchange reaction of a xanthenic derivative in its ground- and excited-state. AB - Buffer-mediated proton exchange reactions of a xanthenic dye were studied in the ground and the excited state by single molecule and bulk fluorescence techniques, respectively. The rate constant obtained supported the uniformity of the process in the ground and the excited state, and the need of adequate character of the buffer species to be able to promote excited-state reactions. PMID- 20023808 TI - Measuring the three-phase contact angle of nanoparticles at fluid interfaces. AB - We report a generic technique to image and study the wettability of spherical nanoparticles adsorbed at liquid surfaces and demonstrate that nanoparticle monolayers can be imprinted at air-water and oil-water interfaces and their three phase contact angle at the original liquid interface can be determined by an atomic force microscopy scan on a replica of the interface; the technique is tested using four different types of nanoparticles, the smallest one having a radius of 37 nm. PMID- 20023809 TI - Manipulating the growth of aqueous semiconductor nanocrystals through amine promoted kinetic process. AB - In the conventional procedure of the preparation of aqueous semiconductor nanocrystals (NCs), the growth of NCs was mainly through the thermodynamics favored Ostwald ripening process. It required additional energy to promote NC growth, such as reflux, hydrothermal method, microwave irradiation, and sonochemical synthesis. Energy-promoted growth usually led to the decomposition of mercapto-ligands and therewith decreased the quality of NCs. Consequently, in this study, the growth of aqueous semiconductor NCs was designed through an amine promoted kinetic process, which efficiently shortened the growth duration and avoided the decomposition of ligands, thus providing a universal method for preparing various aqueous binary and ternary NCs. PMID- 20023810 TI - DFB laser action in a flexible fully plastic multilayer. AB - An all-plastic distributed-feedback flexible multilayer laser (FML), able to keep its lasing characteristics even upon strong bending from planarity, is proposed and the possibility of using this system to make solid-state dye lasers is suggested. PMID- 20023811 TI - Towards understanding of shape formation mechanism of mesoporous silica particles. AB - Growth of even simple crystals is a rather hard problem to describe because of the non-equilibrium nature of the process. Meso(nano)porous silica particles, which are self-assembled in a sol-gel template synthesis, demonstrate an example of shapes of high complexity, similar to those observed in the biological world. Despite such complexity, here we present the evidence that at least a part of the formation of these shapes is an equilibrium process. We demonstrate it for an example of mesoporous fibers, one of the abundant shapes. We present a quantitative proof that the fiber free energy is described by the Boltzmann distribution, which is predicted by the equilibrium thermodynamics. This finding may open up new ground for a quantitative description of the morphogenesis of complex self-assembled shapes, including biological hierarchy. PMID- 20023812 TI - Electron transfer dynamics across self-assembled N-(2-mercaptoethyl) octadecanamide/mycolic acid layers: impedimetric insights into the structural integrity and interaction with anti-mycolic acid antibodies. AB - The integrity and properties of mycolic acid (MA) antigens integrated into a self assembled monolayer (SAM) of N-(2-mercaptoethyl)octadecanamide, (MEODA), on a gold electrode have been interrogated using cyclic voltammetry (CV) and electrochemical impedance spectroscopy (EIS). EIS data showed that Au-MEODA and Au-MEODA-MA behave as microelectrode arrays, with pinholes acting as the microelectrodes that permit electron transport between a redox-active probe in solution and the underlying gold surface. The average radii of the pinholes (r(a)) and half the distance between the centers of the neighbouring pinholes (r(b)), were estimated from EIS using the pore size model, and discussed. Anti-MA antibodies present in a tuberculosis (TB)-infected patient (co-infected with HIV) strongly interact with Au-MEODA-MA showing a rather compact and stable bio complex structure that is virtually defect-free. The electrochemical impedimetric properties associated with the ability of the Au-MEODA-MA to discriminate between TB positive and negative human sera are also discussed. We prove that the Au MEODA and Au-MEODA-MA electrodes, as well as the MA-anti-MA antibody interactions, are characterized with time-constant dispersion, typical of microstructures with grain/grain boundary phases. These crucial physico electrochemical insights into the behaviour of surface-confined MA should provide a useful basis for the design and development of a potential impedimetric immunosensing platform for active tuberculosis. PMID- 20023813 TI - Characterization of Fe sites in Fe-zeolites by FTIR spectroscopy of adsorbed NO: are the spectra obtained in static vacuum and dynamic flow set-ups comparable? AB - The present paper shows a direct comparison of IR spectra of adsorbed NO on two catalyst systems (Fe-silicalite and Fe-ZSM-5), recorded in a flow-through cell (in operando, where NO is carried by an excess of inert gas) and static cell (in situ, where a given pure NO equilibrium pressure is dosed). A progressive NO poly adsorption is observed in the static cell upon increasing the NO equilibrium pressure (from Fe(2+)...(NO)(2) to Fe(2+)...(NO)(3)), while predominantly Fe(2+)...NO adducts are observed in the flow-through cell. By comparing literature spectra, it is shown that these spectral differences are intrinsically inherent to the two different experimental approaches. The two experimental set ups are able to observe preferentially only a part of the total Fe species present on Fe-zeolites. Water contamination experiments employing different experimental conditions (order of dosage, co-dosage, different NO/H(2)O ratios) did not reproduce in the static environment the IR spectra collected with the dynamic set-up. The spectral differences could have a thermodynamic origin and be related to different adsorption enthalpies of mono- and di-nitrosyl complexes of the two Fe(2+) families and to the different NO partial pressure adopted in the two experimental configurations. These considerations have important consequences for the conclusions reached by different research groups on the structure of active Fe sites. PMID- 20023814 TI - Photochemical fabrication of silver nanostructures at the solid-liquid interface using a recyclable photosensitized reduction process. AB - Photochemical synthesis of metal nanostructures through photosensitized reduction of metal sources is a powerful method in a wide variety of environments, because light can selectively promote the desired reaction without damaging the surrounding environment. A drawback of the method is that most organic photosensitizers are disposable and consumed during the photochemical reactions and as a consequence a much larger amount of photosensitizer than that of the metal source is usually required, and therefore, the photosensitized reduction method is wasteful in terms of the sensitizer. In the present work, we propose a new photochemical synthetic method using a recyclable photosensitizer and applied it to fabricate Ag nanostructures at the solid-liquid interface. The photosensitized reduction of Ag ion at the solid-liquid interface resulted in the formation of unique Ag nanostructures (nanowires or plates) without the use of templates or stabilizing agents. The relation between the morphology and several external contributing factors, such as solvent polarity and interaction between the substrate and crystal phase, was discussed. PMID- 20023815 TI - Linear and nonlinear optical properties of triphenylamine-functionalized C60: insights from theory and experiment. AB - In the present study we report on the linear and nonlinear optical properties of C(60)-triphenylamine (TPhA) hybrids. The synthesized materials were prepared following the 1,3-dipolar cycloaddition of azomethine ylides onto the skeleton of C(60) forming the TPhA-based monoadduct, equatorial bis-adduct and dumbell C(60). Complementary spectroscopic techniques, such as NMR, MALDI-TOF-MS, and ATR-IR are applied for the structural characterization of the hybrid materials, while intermolecular electronic interactions are investigated by UV-Vis measurements. In all considered cases, C(60) serves as an acceptor while the triphenylamine unit is chosen as a donor. In order to investigate the dependence of the second order hyperpolarizability on the architecture of the systems (D-A, A-D-A) we use a Z-scan technique employing 532 nm, 35 ps laser pulses. We have found that the total second-order hyperpolarizability of the C(60)-TphA-C(60) system is several times larger than that of TPhA-C(60). The results of experimental measurements are supported by quantum-chemical calculations. PMID- 20023816 TI - Structure and water dynamics of aqueous peptide solutions in the presence of co solvents. AB - We perform neutron diffraction and quasi-elastic neutron scattering (QENS) to probe hydration water structure, and dynamics down to supercooled temperatures, of a concentrated amphiphilic peptide system with the co-solvents glycerol and dimethyl sulfoxide. We find that the kosmotropic co-solvent glycerol preserves the hydration structure near the peptide that is observed in the water solvent alone, that in turn preserves the dynamical temperature trends of two water relaxation processes--one corresponding to a localized relaxation process of the peptide bound surface water and a second relaxation process of the outer hydration layers. By contrast the chaotropic co-solvent, by disrupting the hydration layer near the peptide surface, eliminates the inner hydration layer relaxation process induced by the peptide, to show a single time scale for translational water dynamics. PMID- 20023818 TI - RGD tripeptide onto perfect and grooved rutile surfaces in aqueous solution: adsorption behaviors and dynamics. AB - Molecular dynamics (MD) simulations were performed to investigate the adsorption behavior and dynamics of Arg-Gly-Asp (RGD) tripeptide onto the rutile TiO(2) (110) perfect and grooved surfaces in aqueous solution. The simulation results suggest that, driven by the electrostatic attractions between charged groups of the tripeptide and opposite-type charges of the surface atoms, RGD substitutes the adsorbed water molecules and binds to TiO(2) surface strongly through direct interactions of carboxyl oxygen (O(coo(-))) atoms with nearby titanium atoms in the interface, in agreement with some experimental observations and theoretical data. Once bonded to both perfect and grooved surfaces, RGD tripeptides show a reasonable propensity to remain there with the carboxyl groups providing anchors to the substrate surface, while the amide groups (NH(3)(+) and NH(2)) with larger separations from the attached portions, undergo relatively remarkable fluctuations during the whole simulation time. The trajectories for atom-surface distances, backbone dihedral angles and root-mean-squared deviations from the initial structure have revealed less mobility and more stable adsorption of RGD onto grooved surface than onto perfect surface, which is confirmed again by greater values of adsorption energy for available grooved surfaces. PMID- 20023817 TI - Effects of co-solvents on peptide hydration water structure and dynamics. AB - We evaluate the molecular response of hydration water as a function of temperature and proximity to the surface of the peptide N-acetyl-leucine methylamide (NALMA) when in the presence of the kosmotrope co-solvent glycerol or the chaotrope co-solvent dimethyl sulfoxide (DMSO), using molecular dynamics simulation with a polarizable force field. These detailed microscopic studies complement established thermodynamic analysis on the role of co-solvents in shifting the equilibrium for proteins away from or towards the native folded state. We find that the structure of the water at the peptide interfaces reflects an increase in hydration number in the glycerol solution and a decrease in hydration numbers in the DMSO solution. While the water dynamics around NALMA in the presence of both co-solvents is slower than that observed with the water solvent alone, in the DMSO mixture we no longer measure a separation in water motion time scales at low temperatures as is seen in the pure water solvent, but rather one single relaxation time. In the glycerol, however, we do observe a separation of time scales at low temperatures, supporting the hypothesis that hydration water near a hydrophobic solute evolves on a separate time scale than the extensive hydrogen-bonding network of more bulk-like water. Our simulation studies highlight the differences in the two co-solvent solutions due to the relative frequency of water contacts with the hydrophobic vs. hydrophilic peptide surface, and direct water interactions with the co-solvents. PMID- 20023819 TI - Reformulating time-dependent density functional theory with non-orthogonal localized molecular orbitals. AB - Time-dependent density functional theory (TDDFT) has broad application in the study of electronic response, excitation and transport. To extend such application to large and complex systems, we develop a reformulation of TDDFT equations in terms of non-orthogonal localized molecular orbitals (NOLMOs). NOLMO is the most localized representation of electronic degrees of freedom and has been used in ground state calculations. In atomic orbital (AO) representation, the sparsity of NOLMO is transferred to the coefficient matrix of molecular orbitals (MOs). Its novel use in TDDFT here leads to a very simple form of time propagation equations which can be solved with linear-scaling effort. We have tested the method for several long-chain saturated and conjugated molecular systems within the self-consistent charge density-functional tight-binding method (SCC-DFTB) and demonstrated its accuracy. This opens up pathways for TDDFT applications to large bio- and nano-systems. PMID- 20023820 TI - The detailed balance limit of photochemical energy conversion. AB - Limits and optimization of a solar energy conversion system consisting of a photochemical charge separating unit coupled to an energy storage state are explored by multi-objective genetic algorithms. Pareto fronts were evaluated to obtain information about the ideal parameter combinations, guaranteeing highest efficiency. The light absorbing and charge separating unit is described by a chain of chromophores and electron carriers, connected by Marcus type electron transfer processes. It is coupled to the thermal equilibrium of charge conduction and transport in an energy storage system according to the principle of detailed balance. In addition to our previous findings for an optimal charge separation unit, consisting of a minimum number of charge carriers with adapted recombination and reaction rates, the complete photochemical unit must fulfil further requirements. Low reorganization energies are found to be essential for the initial charge separation steps and can be realized by a low dielectric constant in the local environment. The identified optimal operation rates can be realized by antenna systems adapted to the illumination conditions. For standard solar illumination and a realistic parameter setting energy conversion efficiencies up to 26.8% are predicted, comparable to the limit (31.8%) of ideal single junction semiconductor solar cells. PMID- 20023821 TI - Processing and characterisation of Mo6S2I8 nanowires. AB - One-dimensional nanostructures based on the Mo-S-I system have recently aroused a lot of interest as a viable alternative to the ubiquitous carbon nanotube due to their uniform structure and electronic properties for a given composition. Previous research on the Mo(6)S(3)I(6) and Mo(6)S(4.5)I(4.5) stoichiometries has also shown them to be soluble in common solvents like water, acetone or isopropyl alcohol, and to debundle on dilution. Here, the solubility, debundling and composition of Mo(6)S(2)I(8) nanowires are presented. They were found to be most soluble in dimethylformamide, which retained 47 wt% of a 0.08 gl(-1) nanowire (NW) material dispersion as thin NW bundles after one week. Dispersions of 0.8 gl(-1) and 5 gl(-1) even retained 54 wt% and 66 wt%, respectively. However the NW material was completely insoluble in water, and the surface energy of Mo(6)S(2)I(8) NWs was deduced as 67 mJ m(-2), higher than for other Mo-S-I NWs. UV-vis-NIR spectroscopy showed nanowire peaks familiar from Mo(6)S(3)I(6) and Mo(6)S(4.5)I(4.5) spectra around 1.8 and 2.8 eV, as well as unforeseen ultraviolet peaks at 3.5 and 4.4 eV. These chemical differences suggest an alternate, more strongly bonded structure to that seen for Mo(6)S(3)I(6) and Mo(6)S(4.5)I(4.5) NWs. Films deposited from a range of concentrations were investigated using atomic force microscopy (AFM) to determine bundle diameter distributions. The average diameter and the spread in diameters were found to decrease somewhat with decreasing concentration. However extrapolation gave a finite bundle size at infinite dilution, and an extension of the existing debundling model is proposed to take this into account. To confirm the nominal stoichiometry of Mo(6)S(2)I(8), which does not follow the generic Mo(6)S(x)I(9-x) formula of previous stoichiometries, EDX was carried out. The composition of nanowire bundles was found to be Mo(6)S(2.3)I(8.6), supporting the nominal composition. PMID- 20023822 TI - A DFT periodic study on the interaction between O2 and cation exchanged chabazite MCHA (M = H+, Na+ or Cu+): effects in the triplet-singlet energy gap. AB - O(2) adsorption in proton, sodium and copper exchanged chabazite has been studied using periodic and cluster approaches by means of density functional theory. The Grimme's correction has been used to include the dispersion contribution to B3LYP. Two cation locations have been considered: one with the cation at the six membered ring (MCHA(I)) and the other with the cation at the 8-membered ring (MCHA(IV)). The O(2)-HCHA and O(2)-NaCHA adsorption complexes present a eta(1) O(2) bent coordination. The adsorption energies, which are due to dispersion, are between -15 and -19 kJ mol(-1), in agreement with the experimental values. On the other hand, the O(2) coordination to CuCHA is through a eta(2)-side-on mode with a square planar coordination around the metal center. This structure favors the Cu d -->pi* O(2) charge transfer which becomes the predominant stabilizing factor. The adsorption of singlet states of O(2) in HCHA and NaCHA, modeled with an ONIOM M12T:48T, is of the same nature as that of the ground state, and only the highest in energy (1)Sigma is significantly more stabilized in MCHA than the triplet state by 14 to 24 kJ mol(-1). The adsorption of singlet O(2) in Cu exchanged zeolites presents a higher electron transfer from Cu(+) to O(2) than that calculated for the triplet species and thus both singlet states are stabilized with respect to the ground state O(2). Generally, singlet oxygen appears more attractive to active zeolite models than those calculated with triplet oxygen, indicating a source of oxidative efficiency for designed structures. PMID- 20023823 TI - Influence of phase separating lipids on supported lipid bilayer formation at SiO2 surfaces. AB - The importance of the lipid phase on the formation of supported lipid bilayers (SLBs) via vesicle fusion and on the resulting SLB homogeneity at SiO(2) surfaces has been studied by the quartz crystal microbalance with dissipation (QCM-D) monitoring technique. Physiologically relevant lipid compositions were chosen to correspond to different regions (l(d), l(o) and coexistence of phases) in established phase diagrams of 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC), N-palmitoyl-D-erythro-sphingosylphosphorylcholine (PSM) and cholesterol. For most compositions, SLBs formed through vesicle rupture in a critical-surface coverage dependent manner. Inclusion of PSM and cholesterol into POPC vesicles significantly impaired the vesicle rupture process such that a higher critical concentration of vesicles on the surface was needed before the rupture process started. When increasing the cholesterol content the vesicles formed SLBs containing more defects in the form of intact vesicles adsorbed on the surface up to a point (l(o) phase) where vesicles did not break at all but formed supported vesicular layers. The hampering of vesicle rupture is interpreted in terms of the ability of cholesterol to accommodate vesicle deformation. Experiments using elevated temperatures to alter the lipid membrane into a more fluid phase significantly improved the quality of the SLB showing the importance of both cholesterol content and the lipid phase on SLB homogeneity. PMID- 20023824 TI - On the nature of dense CO adlayers on fcc(100) surfaces: a kinetic Monte Carlo study. AB - We present a kinetic Monte Carlo lattice gas model including top and bridge sites on a square lattice, with pairwise lateral interactions between the adsorbates. In addition to the pairwise lateral interactions we include an additional interaction: an adsorbate is forbidden to adsorb on a bridge site formed by two surface atoms when both surface atoms are already forming a bond with an adsorbate. This model is used to reproduce the low and high coverage adsorption behaviour of CO on Pt(100) and Rh(100). The parameter set used to simulate CO on Pt(100) produces the c(2 x 2)-2t ordered structure at 0.50 ML coverage, a one dimensionally ordered structure similar to the experimentally observed (3 square root(2) x square root(2)) - 2t + 2b structure at 0.67 ML coverage, the c(4 x 2) 4t + 2b ordered structure at 0.75 ML coverage, and the recently reported c(6 x 2) 6t + 4b ordered structure at 0.83 ML coverage. The (5 square root(2) x square root(2)) ordered structure at 0.60 ML coverage is not reproduced by our model. The parameter set used to simulate CO on Rh(100) produces the c(2 x 2)-2t ordered structure at 0.50 ML coverage, a one-dimensionally ordered structure similar to the experimentally observed (4 square root(2) x square root(2)) - 2t + 4b structure at 0.75 ML coverage, and the c(6 x 2)-6t + 4b ordered structure at 0.83 ML coverage. Additionally, the simulated change of top and bridge site occupation as a function of coverage matches the trend in experimental vibrational peak intensities. PMID- 20023825 TI - Solvent polarity effect on intramolecular electron transfer in a corrole naphthalene bisimide dyad. AB - A dyad (C3-NI) based on corrole and naphthalene bisimide has been synthesized and its photoreactivity compared to that of the model component corrole (C3) and naphthalene bisimide (NI) in solvents of different polarity: toluene (TL) and dichloromethane (DCM). The major emitting species in NI solutions, in TL, is identified as a dimeric species (lambda = 470 nm, tau = 2.3 ns) but traces of monomer can also be detected (lambda ca. 390 nm, tau = 40 ps). In DCM the major emitting component is the monomer (lambda = 383 nm, tau = 20 ps) but traces of different aggregates (lambda = 540 and 570 nm, tau = 4.5 and 11 ns) are present. C3 has a fluorescence nearly unaffected by solvent polarity, with a maximum around 655 and a lifetime of 3.5 or 3.8 ns in DCM and TL, respectively. The dyad C3-NI does not appear to be affected by aggregation problems in any of the solvents. Excitation of the imide component in C3-NI (C3-1NI) results in an energy transfer to corrole (1C3-NI) with rate k = 2.0 x 10(11) s(-1) in both solvents. The latter state reacts further via a LUMO-LUMO electron transfer to the naphthalene bisimide yielding the charge separated state C3+-NI- (k = 1.8 x 10(9) s(-1) in TL and k = 3.7 x 10(9) s(-1) in DCM). The same type of reactivity is displayed by direct excitation of the corrole moiety in the dyad to 1C3-NI. C3+-NI- decays with a rate comparable to that of its formation in DCM (k = 4.0 x 10(9) s(-1) in DCM), precluding its accumulation, whereas it decays with a slower rate in TL (k = 7.1 x 10(8) s(-1)). The charge separated state recombines to the ground singlet state; recombination to the triplet state of corrole (excited state at the lowest energy) is in fact excluded on the basis of the experimentally determined triplet yields. The failure of the commonly used methods in the calculation of CS energy levels in apolar solvents is confirmed. PMID- 20023826 TI - EXAFS and XRD characterization of palladium sorbents for high temperature mercury capture from fuel gas. AB - Removal of pollutants such as mercury at elevated temperatures provides improvements in the overall thermal efficiency during the process of coal gasification. The two high temperature sorbents studied were 5 wt% Pd/Al(2)O(3) and 5 wt% Pd/SiO(2): materials shown to have significantly different Hg adsorption capacities. A combination of XRD and EXAFS has been used to characterize the Pd-Hg alloy formed when these Pd-based sorbents were exposed to fuel gas (CO, CO(2), H(2)) containing Hg vapour at 204 degrees C. Significant differences were found in the nature of the alloy formed on the two sorbents following Hg exposure. The Pd/Al(2)O(3) sorbent produced a single homogeneous solid solution of Pd-Hg whilst the silica-supported Pd produced an alloy of varying composition. PMID- 20023827 TI - Phase behaviour and conductivity study on multi-component mixtures for electrodeposition in supercritical fluids. AB - Electrochemistry in supercritical CO(2) (scCO(2)) is difficult because the very low dielectric constant of the fluid restricts the solubility of ionic species and the conductivity of dissolved electrolytes. To overcome this problem to allow us to carry out electrodeposition at macroelectrodes from scCO(2) we have investigated the use of co-solvents and modified electrolyte salts chosen to increase their solubility and dissociation in the supercritical fluid. Here we report results of phase behaviour studies for mixtures of CO(2) with [NBu(n)(4)][BF(4)] and either methanol (CH(3)OH) or acetonitrile (CH(3)CN) as the co-solvent. These show that the solubility of [NBu(n)(4)][BF(4)] is approximately 5 times larger when CH(3)CN is the co-solvent rather than CH(3)OH. Consequently the phase behaviour of the ternary of CO(2)-[NBu(n)(4)][BF(4)]-CH(3)CN was studied in greater detail over a range of compositions. To enhance the conductivity of scCO(2)-CH(3)CN a range of electrolyte salts was synthesised in which the [NBu(n)(4)](+) and/or [BF(4)](-) ion were replaced by different derivatives. Results for the phase behaviour and conductivity of these modified electrolyte salts in scCO(2)-CH(3)CN are reported for several different compositions. We find that increasing the degree of fluorination and size of the ions increases the solubility of the electrolyte salt in scCO(2)-CH(3)CN. Of the 11 electrolytes investigated [NBu(n)(4)][B{3,5-C(6)H(3)(CF(3))(2)}(4)] appears the most suitable for use in scCO(2)-CH(3)CN with a molar conductivity of 22-26 S cm(2) mol(-1) and a maximum measured conductivity of approximately 3 mS cm(-1) for 0.07 M [NBu(n)(4)][B{3,5-C(6)H(3)(CF(3))(2)}(4)] dissolved in scCO(2)-CH(3)CN (molar ratio CH(3)CN : CO(2) approximately 0.12) at 20 MPa and 328.15 K. This is an order of magnitude improvement over similar results for the [NBu(n)(4)][BF(4)] parent. Studies of the conductance as a function of the electrolyte concentration suggest that triple ions make an important contribution to the conductivity of the supercritical fluid. PMID- 20023828 TI - Fast optical vapour sensing by Bloch surface waves on porous silicon membranes. AB - The coupling of optical Bloch surface waves at the truncated end of one dimensional porous silicon photonic crystals is exploited for fast vapour sensing. Self-standing multilayered membranes bound to transparent substrates were fabricated by electrochemical etching and used in an attenuated total reflection configuration to resonantly excite the surface waves and perform real time sensing. PMID- 20023829 TI - Adjustment of the bioresistivity by electron irradiation: self-assembled monolayers of oligo(ethyleneglycol)-terminated alkanethiols with embedded cleavable group. AB - The bioresistivity of protein-repelling films, such as e.g. oligoethyleneglycol (OEG) bearing self-assembled monolayers (SAMs), can be adjusted by electron irradiation. We have studied the effect of an embedded irradiation-sensitive functional group (so called "weak link") on the irradiation sensitivity of such films. As test systems, we used two OEG-substituted alkanethiolate SAMs with different lengths of the OEG chain and a sulfone group serving as a "weak link" moiety; this group was embedded between the OEG and alkyl chains of the target molecules. The expected cleavage of the molecular chains at the predetermined "weak link" position was found to be accompanied by direct damage of the OEG matrix, with a dominance of the latter process in the case of a long OEG chain. At the same time, in the case of short OEG chain, the insertion of sulfone group resulted in a noticeable gain in the irradiation sensitivity of the respective SAMs and, consequently, in enhanced tunability of their protein-adhesive properties. These films, along with other OEG-substituted SAMs and polymer layers with and without an embedded weak link moiety, can be used as primary matrices for the fabrication of different protein patterns by electron beam lithography. PMID- 20023830 TI - A proton NMR relaxation study of water dynamics in bovine serum albumin nanoparticles. AB - Water dynamics and compartmentation in glutaraldehyde cross-linked bovine serum albumin nanoparticles have been investigated by an integrated nuclear magnetic resonance (NMR) protocol based on water relaxation times and self-diffusion coefficients measurements. Multi-exponentially of water relaxation curves has been accounted for according to a diffusive and chemical exchange model (see B. P. Hills, S. F. Takacs and P. S. Belton, Mol. Phys., 1989, 67(4), 903, and Mol. Phys., 1989, 67(4), 913; E. Brosio, M. Belotti and R. Gianferri, in Food Science and Technology: New Research, ed. L. V. Greco and M. N. Bruno, Nova Science Publishers, Hauppauge (NY), 2008) that made it possible to single out water molecules in the molecular spaces in the interior of albumin nanoparticles, in the meso-cavities formed by packed nanoparticles and in the meniscus on top of the nanoparticles suspension. A quantitative rationalization of T(2) values of water different components allowed morphological information to be acquired as for the size of water filled compartments, while self-diffusion coefficient measurements of water excess or fluxed packed nanoparticles suspensions are describers of transport properties of soft biomaterials. The paper reports an NMR approach that can be seen as a general and relevant method to characterize excess water-swollen soft biomaterials. PMID- 20023831 TI - Folding oligomers of difluorinated thienylfurans: a DFT study. AB - Density functional theory calculations were performed on a series of difluorinated-thienylfuran oligomers containing up to six repeat units. For any given oligomer, a building unit was chosen to be either in the transoid or in the cisoid conformation. The oligomers are found to be more stable in the all-cisoid conformation as a result of intramolecular F...H hydrogen bonding interactions, as well as stacking pi-pi interactions for the larger oligomers. The relative stability increases markedly with the size of the oligomers, allowing for the adoption of well-defined folding helical patterns for systems with at least five repeat units. Conveniently designed isodesmic reactions show that folding of the oligomers is a thermodynamically favorable process. PMID- 20023833 TI - Mechanically bonded macromolecules. AB - Mechanically bonded macromolecules constitute a class of challenging synthetic targets in polymer science. The controllable intramolecular motions of mechanical bonds, in combination with the processability and useful physical and mechanical properties of macromolecules, ultimately ensure their potential for applications in materials science, nanotechnology and medicine. This tutorial review describes the syntheses and properties of a library of diverse mechanically bonded macromolecules, which covers (i) main-chain, side-chain, bridged, and pendant oligo/polycatenanes, (ii) main-chain oligo/polyrotaxanes, (iii) poly[c2]daisy chains, and finally (iv) mechanically interlocked dendrimers. A variety of highly efficient synthetic protocols--including template-directed assembly, step-growth polymerisation, quantitative conjugation, etc.--were employed in the construction of these mechanically interlocked architectures. Some of these structures, i.e., side-chain polycatenanes and poly[c2]daisy chains, undergo controllable molecular switching in a manner similar to their small molecular counterparts. The challenges posed by the syntheses of polycatenanes and polyrotaxanes with high molecular weights are contemplated. PMID- 20023834 TI - Fabrication of biomolecular devices via supramolecular contact-based approaches. AB - This tutorial review provides an outlook on contact-based fabrication methods suitable for biomolecular platforms. Contact-based methods have emerged in response to serial and expensive fabrication techniques that built devices in a serial way, typically point by point or region by region. The review surveys the biological applications of microcontact printing and affinity contact printing. There is a special focus on DNA printing methods harnessing the supramolecular interactions between two complementary DNA strands. PMID- 20023835 TI - Fibre optic microarrays. AB - This tutorial review describes how fibre optic microarrays can be used to create a variety of sensing and measurement systems. This review covers the basics of optical fibres and arrays, the different microarray architectures, and describes a multitude of applications. Such arrays enable multiplexed sensing for a variety of analytes including nucleic acids, vapours, and biomolecules. Polymer-coated fibre arrays can be used for measuring microscopic chemical phenomena, such as corrosion and localized release of biochemicals from cells. In addition, these microarrays can serve as a substrate for fundamental studies of single molecules and single cells. The review covers topics of interest to chemists, biologists, materials scientists, and engineers. PMID- 20023837 TI - Development and applications of tethered bis(8-quinolinolato) metal complexes (TBOxM). AB - The tethered bis(8-quinolinolato) metal (TBOxM) complexes have found practical use in many asymmetric reactions. The development of highly enantioselective reactions using TBOxM is surveyed. General reaction schemes and examples using these highly active and selective catalysts are given and discussed in this tutorial review. PMID- 20023836 TI - Responsive magnetic resonance imaging contrast agents as chemical sensors for metals in biology and medicine. AB - This tutorial review highlights progress in the development of responsive magnetic resonance imaging (MRI) contrast agents for detecting and sensing biologically relevant metal ions. Molecular imaging with bioactivatable MRI indicators offers a potentially powerful methodology for studying the physiology and pathology of metals by capturing dynamic three-dimensional images of living systems for research and clinical applications. This emerging area at the interface of inorganic chemistry and the life sciences offers a broad palette of opportunities for researchers with interests ranging from coordination chemistry and spectroscopy to supramolecular chemistry and molecular recognition to metals in biology and medicine. PMID- 20023838 TI - Bright functional rotaxanes. AB - As a hot topic in research, various rotaxanes continue to be constructed. This tutorial review focuses mainly on bright rotaxanes with functional properties, which have been developed in recent years: fluorescent rotaxanes, rotaxanes as logic gates and information storage devices, gelation of rotaxanes, rotaxanes on solid surfaces, sensory rotaxanes, as well as rotaxanes related to biology and so on. Novel synthetic protocols towards several functional rotaxanes are also illustrated. PMID- 20023839 TI - Hydrogen generation from formic acid and alcohols using homogeneous catalysts. AB - This tutorial review describes recent progress in the development of homogeneous catalytic methodology for the direct generation of hydrogen gas from formic acid and alcohols. PMID- 20023840 TI - Rearrangement of beta-amino alcohols via aziridiniums: a review. AB - This tutorial review focuses on the rearrangement of beta-amino alcohols via aziridinium intermediates. It covers the literature from 1947 to January 2009 (55 references). The rearrangement of beta-amino alcohols can be performed by activation of the hydroxy group followed by the addition of nucleophiles (Nu). In most examples, an aziridinium intermediate is involved in the rearrangement. The ratio of amines resulting from the attack of nucleophiles at either the C-1 or C 2 position of the aziridinium intermediate, depends on the nature of the nucleophiles and the R(2) substituent. In some cases, solvent as well as temperature can influence the ratio of amines. PMID- 20023841 TI - Chemistry and materials options of sustainable carbon materials made by hydrothermal carbonization. AB - The production of functional nanostructured materials starting from cheap natural precursors using environmentally friendly processes is a highly attractive subject in material chemistry today. Recently, much attention has been focused on the use of plant biomass to produce functional carbonaceous materials, encompassing economic, environmental and social issues. Besides the classical route to produce activated carbons from agricultural side products, the hydrothermal carbonization (HTC) process shows clear advantages in that it can generate a variety of cheap and sustainable carbonaceous materials with attractive nanostructure and functionalization patterns for a wide range of applications. In this tutorial review we present the latest developments in this traditional but recently invigorated technique. It will be shown that HTC does not only access carbonaceous materials under comparatively mild hydrothermal conditions, but also replaces the more technical and structurally well-defined charring by a controlled chemical process. It will be shown that this makes it possible to tailor the final structure with the tools of colloid and polymer science, leading to very different morphologies with miscellaneous applications, including modern carbon nanocomposites and hybrids. PMID- 20023842 TI - Natural and artificial peptide motifs: their origins and the application of motif programming. AB - In this tutorial review, I discuss the nature and application of peptide motifs. Motifs are usually identified through analysis of the sequence of natural proteins and are linked to particular biological functions, though the association between a motif and its function is only speculative in some cases. In other cases, however, the transplantability and functional independence of motifs have been experimentally proven, providing us with the opportunity to use those motifs as programming units for biotechnological application. In addition to natural motifs, peptide aptamers created using in vitro evolution systems can also serve as motif units. The associated functions of these artificial motifs are related to their binding ability. Numerous binders against both natural biomolecules and inorganic materials have been created from peptide phage systems. By programming these natural and artificial motifs, artificial proteins with the potential to contribute to medical diagnosis and treatment, nanotechnology, and various areas of basic science have been created. In addition, the transplantability and functional independence of motifs provide insight into the nature of protein evolution. PMID- 20023843 TI - Sensors for the optical detection of cyanide ion. AB - This tutorial review focuses on recent developments arising from studies of optical sensors for cyanide ions, which are categorized by approaches involving cyanide selective receptors, the utilization of metal coordinated complexes, and chemodosimeters. PMID- 20023844 TI - Supramolecular n/p-heterojunction photosystems with oriented multicolored antiparallel redox gradients (OMARG-SHJs). AB - Directional electron and hole transport is essential in photosynthesis. Applied to molecular optoelectronics such as organic solar cells, these lessons from nature call for oriented supramolecular n/p-heterojunctions (SHJs) that contain various chromophores and antiparallel redox gradients (OMARGs). In this tutorial review, we summarize recent progress made toward the construction of OMARG-SHJs. This conceptually innovative twist added to a timely topic should appeal to the synthetic organic, supramolecular, biological, physical, analytical and materials chemist as well as to the expert in energy and environmental sciences. PMID- 20023845 TI - Functional evolution on the assembled DNA template. AB - After more than twenty years of effort, using DNA to fabricate addressable nanostructures is now a well-established technology. In this tutorial review, we attempt to present an overview of the applications of DNA templates, including its biological significance, the directing of chemical reactions at the molecular level, as well as the placing of nanoparticles and proteins in position. PMID- 20023846 TI - Hierarchical structures made of proteins. The complex architecture of spider webs and their constituent silk proteins. AB - Biopolymers fulfil a variety of different functions in nature. They conduct various processes inside and outside cells and organisms, with a functionality ranging from storage of information to stabilization, protection, shaping, transport, cellular division, or movement of whole organisms. Within the plethora of biopolymers, the most sophisticated group is of proteinaceous origin: the cytoskeleton of a cell is made of protein filaments that aid in pivotal processes like intracellular transport, movement, and cell division; geckos use a distinct arrangement of keratin-like filaments on their toes which enable them to walk up smooth surfaces, such as walls, and even upside down across ceilings; and spiders spin silks that are extra-corporally used for protection of offspring and construction of complex prey traps. The following tutorial review describes the hierarchical organization of protein fibers, using spider dragline silk as an example. The properties of a dragline silk thread originate from the strictly controlled assembly of the underlying protein chains. The assembly procedure leads to protein fibers showing a complex hierarchical organization comprising three different structural phases. This structural organization is responsible for the outstanding mechanical properties of individual fibers, which out-compete even those of high-performance artificial fibers like Kevlar. Web-weaving spiders produce, in addition to dragline silk, other silks with distinct properties, based on slightly variant constituent proteins--a feature that allows construction of highly sophisticated spider webs with well designed architectures and with optimal mechanical properties for catching prey. PMID- 20023847 TI - Stereocontrolled ring-opening polymerization of cyclic esters: synthesis of new polyester microstructures. AB - Synthesis of aliphatic polyesters has been studied intensively due to their biocompatible and biodegradable properties and their potential applications in medical and agricultural fields. There has been particular emphasis over the past decade on the synthesis of discrete, well-characterized complexes that are active polymerization initiators for the ring-opening polymerization (ROP) of lactide (LA) and beta-butyrolactone (BBL) to give, respectively, poly(lactide) (PLA) and poly(3-hydroxybutyrate) (PHB). These recent advances in catalyst design have led to a variety of polyester microstructures. This tutorial review focuses on the use of metal-based complexes for the stereoselective ROP of rac-LA and rac-BBL. PMID- 20023848 TI - Smart molecules at work--mimicking advanced logic operations. AB - Molecular logic is an interdisciplinary research field, which has captured worldwide interest. This tutorial review gives a brief introduction into molecular logic and Boolean algebra. This serves as the basis for a discussion of the state-of-the-art and future challenges in the field. Representative examples from the most recent literature including adders/subtractors, multiplexers/demultiplexers, encoders/decoders, and sequential logic devices (keypad locks) are highlighted. Other horizons, such as the utility of molecular logic in bio-related applications, are discussed as well. PMID- 20023849 TI - Lanthanide luminescence for functional materials and bio-sciences. AB - Recent startling interest for lanthanide luminescence is stimulated by the continuously expanding need for luminescent materials meeting the stringent requirements of telecommunication, lighting, electroluminescent devices, (bio )analytical sensors and bio-imaging set-ups. This critical review describes the latest developments in (i) the sensitization of near-infrared luminescence, (ii) "soft" luminescent materials (liquid crystals, ionic liquids, ionogels), (iii) electroluminescent materials for organic light emitting diodes, with emphasis on white light generation, and (iv) applications in luminescent bio-sensing and bio imaging based on time-resolved detection and multiphoton excitation (500 references). PMID- 20023850 TI - The chemistry of graphene oxide. AB - The chemistry of graphene oxide is discussed in this critical review. Particular emphasis is directed toward the synthesis of graphene oxide, as well as its structure. Graphene oxide as a substrate for a variety of chemical transformations, including its reduction to graphene-like materials, is also discussed. This review will be of value to synthetic chemists interested in this emerging field of materials science, as well as those investigating applications of graphene who would find a more thorough treatment of the chemistry of graphene oxide useful in understanding the scope and limitations of current approaches which utilize this material (91 references). PMID- 20023851 TI - Amphiphilic peptides and their cross-disciplinary role as building blocks for nanoscience. AB - Peptides are particularly attractive as molecular building blocks in the bottom up fabrication of supramolecular structures based on self-assembly and have potential in many important applications in the fields of biotechnology and bioengineering. In the first part of this critical review the main categories of peptide-based amphiphiles will be discussed by showing some relevant examples, which demonstrate the importance of amphiphilic peptides as molecular building blocks for nanostructures. In the second part of this review we will review the cross-disciplinary role of peptide-based supramolecular nanoarchitectures ranging from chemistry to biology, medicine, materials science, and engineering through discussing several examples of applied nanomaterials (216 references). PMID- 20023852 TI - Discotic nematic liquid crystals: science and technology. AB - The nematic phase of discotic liquid crystals, although rarely observed, has made very significant progress over the past three decades since their discovery. It has made its way from a mere scientific curiosity to application in commodities. The negative birefringence films formed by polymerized nematic discotic liquid crystals have been commercialized as compensation films to enlarge the viewing angle and enhance the contrast ratio of commonly used twisted nematic liquid crystal displays. High strength and high performance carbon fibers for industrial applications have been obtained from the carbonaceous mesophase and a liquid crystal display device with wide and symmetrical viewing angle has been demonstrated by using discotic nematic liquid crystals. Discotic films with patterned colours have been obtained from cholesteric lyo-mesophases of discotic liquid crystals. Various molecular architectures have been designed and synthesized to exhibit the discotic nematic phase over a wide range of temperature. This critical review focuses on the synthesis and physical properties of these fascinating materials. It deals with the structure of various nematic phases, different discotic cores exhibiting the nematic phase, novel designing and transition temperature engineering principles, alignment and physical properties, and finally the application of discotic nematic LCs as the active switching component and as optical compensation films for widening the viewing angle and contrast ratio of liquid-crystal display devices (98 references). PMID- 20023853 TI - Controlled polymer synthesis--from biomimicry towards synthetic biology. AB - The controlled assembly of synthetic polymer structures is now possible with an unprecedented range of functional groups and molecular architectures. In this critical review we consider how the ability to create artificial materials over lengthscales ranging from a few nm to several microns is generating systems that not only begin to mimic those in nature but also may lead to exciting applications in synthetic biology (139 references). PMID- 20023854 TI - Green chemistry: principles and practice. AB - Green Chemistry is a relatively new emerging field that strives to work at the molecular level to achieve sustainability. The field has received widespread interest in the past decade due to its ability to harness chemical innovation to meet environmental and economic goals simultaneously. Green Chemistry has a framework of a cohesive set of Twelve Principles, which have been systematically surveyed in this critical review. This article covers the concepts of design and the scientific philosophy of Green Chemistry with a set of illustrative examples. Future trends in Green Chemistry are discussed with the challenge of using the Principles as a cohesive design system (93 references). PMID- 20023855 TI - Biocatalytic carboxylation. AB - Dwindling petroleum feedstocks and increased CO(2)-concentrations in the atmosphere currently open the concept of using CO(2) as raw material for the synthesis of well-defined organic compounds. In parallel to recent advances in the chemical CO(2)-fixation, enzymatic (biocatalytic) carboxylation is currently being investigated at an increased pace. On the one hand, this critical review provides a concise overview on highly specific biosynthetic pathways for CO(2) fixation and, on the other hand, a summary of biodegradation (detoxification) processes involving enzymes which possess relaxed substrate specificities, which allow their application for the regioselective carboxylation of organic substrates to furnish the corresponding carboxylic acids (145 references). PMID- 20023856 TI - Polypeptide-polymer bioconjugates. AB - Creating bioconjugates by combining polymers with peptides and proteins is an emerging multidisciplinary field of research that has enjoyed increased attention within the scientific community. This critical review provides an overview of the strategies employed for the construction of these materials and will highlight the underlying synthetic methods used. This review is therefore relevant for chemists, material scientists and chemical biologists facing the challenge of constructing polypeptide-polymer bioconjugates in a controlled fashion (269 references). PMID- 20023857 TI - Dynamic control over cell adhesive properties using molecular-based surface engineering strategies. AB - In complex organisms, cells are often dependent on their extracellular matrix (ECM) for structural integrity, the mechanical properties of tissues, and for signaled regulation of cellular processes including adhesion, migration, growth, secretion, gene expression and apoptosis. Achieving dynamic control, i.e. by using an external stimulus, over the interactions between cells and artificial interfaces holds considerable promise in tissue engineering, medicine, cell biology and immunology. For example, improved spatial control over cell-surface interaction is potentially useful in the design of cell-based screening devices. Dynamic control over SAMs for cell adhesion provides an additional handle to direct and study the attachment of cells to surfaces, e.g., in studying cell spreading from a predetermined pattern in order to screen the cytotoxicity of drug candidates. However, 'reversible' control of cell adhesion onto substrates is an area that is still in its infancy. In this critical review recent developments in cell adhesion of mammalian cells to SAM-modified surfaces, the physical properties of which can be controlled by an external stimulus, e.g. by light, electrochemistry, etc., are discussed (118 references). PMID- 20023858 TI - Cyclic and acyclic oligo(N2O2) ligands for cooperative multi-metal complexation. AB - This review describes the design and synthesis of hosts based on salen or related N(2)O(2) ligands, which are useful for the development of cooperative metal complexation systems. A key to the design of the N(2)O(2)-based metallohosts is the negative charges of the phenoxo groups, which further coordinate to metal ions in a bridging fashion. In particular, the integration of two or more N(2)O(2) coordination sites is effective for the construction of metallohosts. A variety of cyclic and acyclic oligo(N(2)O(2)) compounds were designed and synthesized. They show unique complexation behavior due to the integrated coordination sites. For examples, cyclic tris(N(2)O(2)) ligands formed novel cluster complexes and acyclic bis(N(2)O(2)) ligands bind alkaline earth and rare earth metal ions in the central recognition site via metal exchange. The suitable arrangement of the phenoxo moieties contributes to the preorganization for the cooperative metal complexation, producing metallocyclic or metallohelical structures. PMID- 20023859 TI - Organopalladium and platinum chemistry in oxidising milieu as models for organic synthesis involving the higher oxidation states of palladium. AB - This perspective focuses on the higher oxidation state (III, IV) organometallic chemistry of palladium involving a range of strong oxidants, with consideration of platinum chemistry where it is informative for the evaluation of structure and mechanism. Particular emphasis is placed on hypervalent iodine reagents, halogens and related oxidants of intense current interest in organic synthesis, together with linkages in concepts between this chemistry and recent advances in studies of diaryl disulfides, diaryl diselenides, and diaryl peroxides as oxidants. PMID- 20023860 TI - Template control in ionothermal synthesis of aluminophosphate microporous materials. AB - Pentaerythritol/quaternary ammonium eutectic mixture as an ionic solvent: the quaternary ammonium cations can be easily controlled to exert their structure directing effect in ionothermal synthesis of aluminophosphate microporous materials. PMID- 20023861 TI - A thiazoline-containing cobalt(II) complex based colorimetric fluorescent probe: "turn-on" detection of fluoride. AB - A new thiazoline-containing cobalt(II) complex displayed colorimetric and fluorescent "turn-on" selectivity and sensitivity toward fluoride anions compared to the other anions investigated through color changes from pink to blue and changes in the emission spectra at approximately 570 nm when excited at 520 nm. PMID- 20023862 TI - Controlling the molecular aggregation. An amphiphilic Schiff-base zinc(II) complex as supramolecular fluorescent probe. AB - A designed amphiphilic Schiff-base bis(salicylaldiminato)zinc(II) complex, 1, forms defined aggregates in dilute solutions of non-coordinating solvents. The switching to the monomer can be driven by addition of a coordinating species and involves a dramatic enhancement of the fluorescence emission. PMID- 20023863 TI - A unique type of a dicobalt cage templated by a weakly coordinated hexafluorophosphate anion: design, structure and solid-state NMR investigations. AB - A dinuclear cobalt capsule [PF(6) subset(CH(3)CN)(2)(Co(2)L(4))][PF(6)](3) (2), (L = bis-benzimidazole-1,3-phenylene) has been prepared using a self-assembly approach. This unique class of cage is perfectly designed to encapsulate weakly coordinated anions, i.e. BF(4)(-) and PF(6)(-)via an Manion weak interaction as illustrated by X-ray and solid-state NMR investigations. Selective anion binding for BF(4)(-) over PF(6)(-) is also observed. PMID- 20023864 TI - Hydride-induced cleavage of C-O bond in pyran ring of 2-pyranylidene carbene complexes: mechanism and synthetic application. AB - Hydride-induced cleavage of the C-O bond in the pyran ring followed by ring closure of unsaturated acylmetalates and reductive elimination was achieved. PMID- 20023865 TI - Conformational analysis of PEt3 and P(OMe)3 in metal complexes. AB - The conformations of the archetypal acyclic phosphorus ligands PEt(3) and P(OMe)(3) are classified on the basis of the observation that torsions about the P-C (or P-O) bonds show favoured conformations lying close to gauche (+/-60 degrees) or anti values (180 degrees). Analysis of the symmetry of the conformation space defined by the three M-P-X-C (X = CH(2) or O) torsion angles (t(1-3)) implies the existence of seven unique conformer types (A (aaa), B (g(+)g(+)g(+)), C (ag(+)g(+)), D (aag(+)), E (g(-)ag(+)), F (ag(-)g(+)), G (g( )g(+)g(+)) and their symmetry equivalents) arising from the combinations of g(-), g(+) and a torsions. These conformers are observed in 1972 M-PEt(3) and 735 M P(OMe)(3) fragments from crystal structures of metal complexes in the CSD following the popularity sequence: F > C >> D > B > G > E >> A for M-PEt(3); and: C > D > F >> E >> A, B, G. for M-P(OMe)(3). Pathways for low-energy interconversion of these conformers, dominated by single chain flip routes, are readily inferred for M-P(OMe)(3). The conformers of M-PEt(3) are apparently less readily interconverted. The popularity of conformations is only loosely related to the energies of these conformations as calculated by DFT or MM methods for two , four- (square planar) and six-coordinate metal complexes of these ligands (and free PEt(3) and P(OMe)(3)). It would appear that the conformational preferences observed are determined by a balance between intra-ligand effects (repulsion between chains and avoidance of syn-pentane-like); inter-ligand effects (repulsions between gauche substituents at P and cis co-ligands notably when the coordination number at the metal is high); and residual anomeric effects (weakly favouring anti conformations in P(OMe)(3) species). PMID- 20023866 TI - Synthesis, structural characterization and in vitro cytotoxicity of new Au(III) and Au(I) complexes with thioamides. AB - The reactions of tetrachloroauric(III) acid (HAuCl4) with the thioamides; 2 mercapto-benzothiazole (mbztH) and 5-ethoxy-2-mercapto-benzimidazole (EtmbzimH) lead to the desulfuration of the ligands and the formation of the ionic complexes {[AuCl4]- [bztH2]+} (1), and {[AuCl4]- [EtbzimH2]+ (H2O)} (2) (where bztH2+ and EtbzimH2+ are the desulfurated cations of the starting ligands). The reaction of HAuCl4 with 2-mercapto-nicotinic acid (mnaH2), however results in the formation of 2-sulfonate-nicotininc acid (C6H5NO5S) (3) with the simultaneous oxidation of the sulfur atom. On the other hand, the reactions of the gold(I) complex [Au(tpp)Cl] (4) (tpp = triphenylphosphine (Ph3P)) with the thioamides; 2-mercapto thiazolidine (mtzdH), 2-mercapto-benzothiazole (mbztH) and 5-chloro-2-mercapto benzothiazole (ClmbztH) in the presence of potassium hydroxide resulted in the formation of the gold(I) complexes of formulae [Au(tpp)(mtzd)] (5), [Au(tpp)(mbzt)] (6) and [Au(tpp)(Clmbzt)] (7) without ligand desulfuration. All complexes have been characterized by elemental analysis, FT-IR, far-FT-IR,1H-NMR, spectroscopic techniques and X-Ray crystallography. The electrochemical behavior of 1, 2 and 4-7 complexes and the ligands EtmbzimH, mbztH and mnaH2 was also studied in acetonitrile and DMF using cyclic voltammetry. The results are in support of a mechanism of desulfuration of the ligands by Au(III), involving a first oxidation of S to -SO3-, followed by a C-S bond cleavage. This is also supported by PM6 calculations of bond dissociation energies of the various compounds involved. Complexes 1, 2 and 4-7 were tested for in vitro cytotoxicity against leiomyosarcoma cells and the results are discussed in relation with the geometry of the complexes and compared with those of cisplatin and other metals. Complexes 1 and 5 showed higher activity than that of cisplatin, while HAuCl4 was inactive against sarcoma cells. PMID- 20023867 TI - Structural modification of rhodamine-based sensors toward highly selective mercury detection in mixed organic/aqueous media. AB - In virtue of the sulfurphilic nature of Hg(2+), three new sensors RN1, RN2 and RST1 that combine a thiophene group and one or two rhodamine choromophores, or a thiospirolactam rhodamine chromophore, were designed and prepared for the selective detection of Hg(2+) in aqueous media, respectively. These sensors all displayed good brightness and fluorescence enhancement following Hg(2+) coordination with limits of detection for Hg(2+) at the ppb level. Thus, they have the potential for distinguishing between safe and toxic levels of inorganic mercury in drinking water. RN1 exhibited chromogenic and fluorogenic selectivity over alkali, alkaline earth metals, divalent first-row transition metal ions as well as heavy metals, but the presence of Cu(2+) had a small but significant influence on the absorption detection of Hg(2+). Compared to RN1, the introduction of sufficient sulfur atoms could increase the binding capability of RST1 towards Hg(2+) relative to the sensor RN1, but decrease its Hg specific ability. The existence of some heavy and transition metal ions, such as Pb(2+), Ag(+), Cu(2+) enhance the silent absorption spectra of RST1. Spectral evidence and X-ray structural investigations of the mercury complex revealed a possible 1:2 complexation behaviour between the Hg(2+) ion and the sensor RN1 or RST1. Sensor RN2 which contains two rhodamine carboxhydrazone arms exhibited better selectivity, compared to those of RN1 and RST1. The addition of Cu(2+) only caused a small interference for the absorption detection of Hg(2+) under the same conditions, demonstrating the efficiency of the robust bis-chelating mode with regard to the selectivity for Hg(2+). PMID- 20023868 TI - Systematic investigation of an array of TCNQ lanthanide complexes: synthesis, structure and magnetic properties. AB - An array of TCNQ (TCNQ = 7,7,8,8-tetracyano-p-quinodimethane) lanthanide complexes, namely, [Ln2(TCNQ)4(H2O)10(EtOH)2][2TCNQ] x xH2O [Ln = La (1, x = 5), Pr (2, x = 3)], [Ln2(TCNQ)4(H2O)10][2TCNQ] x yH2O [Ln = Gd (3, y = 6), Dy (4, y = 5)] and [Ln(TCNQ)2(H2O)6][TCNQ] x H2O x MeOH [Ln = Er (5), Lu (6)], have been synthesized by salt-elimination reactions. X-ray crystallographic analysis indicates that 1-6 are discrete ion complexes. Cations of 1-4 adopt a dimer structure, while cations of 5 and 6 display a monomer structure. The magnetic study proposes that an antiferromagnetic interaction between TCNQ radicals exists in 1-6. The depopulation of the Stark levels for 2 and 5 leads to a continuous decrease in mT when the samples are cooled from 300 K to 2 K. The desolvated 3, namely, 3a exhibits a unique magnetic conversion from the magnetic disordering to the magnetic ordering at 4 K. 4 shows a ferromagnetic coupling between paramagnetic centers. PMID- 20023869 TI - Butterfly and rhombus structures for binuclear cobalt carbonyl sulfur and phosphinidene complexes of the type Co2(CO)6E2 (E = S, PX). AB - Theoretical studies on Co(2)(CO)(6)(PX)(2) derivatives (X = H, Cl, OH, OMe, NH(2), NMe(2)) predict the lowest energy structures to be butterfly structures containing five two-electron two-center bonds in the central Co(2)P(2) unit. Among these butterfly structures the energy increases as the unique bond forming the "body" of the butterfly changes from Co-Co to Co-P and then P-P. Higher energy rhombus structures are also found for Co(2)(CO)(6)(PX)(2) with only Co-P bonds in the Co(2)P(2) framework without any Co-Co or P-P bonds. In addition, for Co(2)(CO)(6)(POR)(2) (R = H, Me) still higher energy "diphosphine" structures are also found containing only three rather than four Co-P bonds, one P-P bond, and no Co...Co bond. For the isoelectronic Co(2)(CO)(6)S(2) a rhombus structure is competitive in energy with the butterfly structures with five structures lying within approximately 4 kcal/mol thereby predicting a fluxional system. A tetrahedrane structure was not found for Co(2)(CO)(6)S(2) in contrast to the tetrahedrane structure known experimentally for the related Fe(2)(CO)(6)S(2) with one less electron per metal atom. PMID- 20023870 TI - Homogeneous and heterogeneous catalytic oxidation of sulfides by H2O2 over zinc(II) compounds. AB - It has been recently shown that zinc compounds are effective catalysts for the oxidation of alkyl aryl sulfides to the corresponding sulfoxides in the presence of hydrogen peroxide. In this paper, we have investigated homogeneous and heterogeneous catalytic oxidation of sulfides by H(2)O(2) over Zn(NO(3))(2) x 6 H(2)O and the metal-organic porous material [Zn(2)(bdc)(L-lac)(dmf)] x DMF (where H(2)bdc = p-benzenedicarboxylic acid, H(2)lac = lactic acid), respectively. The experimental data can be explained by the proposed catalytic cycle which includes the activation of H(2)O(2) via coordination to Zn(II) ions followed by oxygen transfer step. In homogeneous conditions, the presence of a large amounts of H(2)O(2) results in the coordination of two molecules of hydrogen peroxide to Zn(II), so that sulfone is formed via transfer of two oxygen atoms from Zn(H(2)O)(4)(H(2)O(2))(2)(2+) active species. Contrary to the homogeneous system, the use of [Zn(2)(bdc)(L-lac)(dmf)] x DMF as catalyst does not lead to the formation of sulfone in the initial period of reaction. This is consistent with the proposed catalytic cycle of sulfoxidation as each Zn(II) center in the crystalline framework is able to activate only one H(2)O(2) molecule. Our investigations indicate that the sorption and activation of H(2)O(2) molecules by microporous framework [Zn(2)(bdc)(L-lac)(dmf)] occur faster than sulfide sorption and oxygen transfer. PMID- 20023871 TI - The reaction of tin(IV) iodide with phosphines: formation of new halotin anions. AB - The reaction of SnI(4) with each of a primary, secondary and tertiary phosphine has been investigated and in none of the cases are simple adducts formed. With Cy(3)P, [Cy(3)PI](+) salts of both [SnI(3)](-) (1) and [SnI(5)](-) (2) are isolated arising from reactions involving both reduction at tin and halogen transfer. With the secondary and primary amines Ph(2)PH and CyPH(2), respectively, additional HI elimination reactions occur and the salts [Ph(2)PH(2)](+)(6)[Sn(3)I(12)](6-) (3), [Ph(2)PH(2)](+)(2)[SnI(6)](2-) (4) and [CyPH(3)](+)(2)[SnI(4)](2-) (5) have been isolated. Compounds 1-5 have been characterised crystallographically. PMID- 20023872 TI - Coordinative flexibility in an acyclic bis(sulfonamide) ligand. AB - The new acyclic and potentially heptadentate dinucleating ligand, 2,6-bis(N,N-bis (2-pyridylmethyl)-sulfonamido)-4-methylphenolato (bpsmp(-)) contains two tertiary sulfonamide groups. The sulfonamide donors permit a greater degree of control over the accessibility of mono versus dinuclear complexes compared to their closely related amine-containing counterparts, on account of their relatively weaker donor properties. A series of air-stable dinuclear complexes of Co(II), Mn(II) and Cu(II) containing two auxiliary acetate ligands have been prepared. The absence of acetate in reaction mixtures containing Co(II) and Mn(II) led to mononuclear complexes, with water ligands completing the coordination spheres of the metal ions, even in the presence of large excess of the metal ions. Thus, bridging acetate ligands appear to stabilise the dinuclear structures for the relatively labile Co(II) and Mn(II) ions. A mononuclear complex of V(IV)=O was isolated even in the presence of acetate, possibly because the oxyl groups on each V(IV) prevent formation of a bis-acetato-bridged complex. Reaction of one equivalent of CuCl(2) with bpsmpH led to isolation of two different mononuclear complexes, dependent on the identity of the solvent. The phenol group is coordinated in only one of these complexes. A dinuclear Cu(II) complex was isolated when two equivalents of the metal salt were used in the reaction. PMID- 20023873 TI - Syntheses, structures, and photoluminescence of 1-D lanthanide coordination polymers. AB - Five new lanthanide 1-D coordination polymers are reported which are formed from flexible salen type Schiff-base ligands H(2)L and H(2)L' (H(2)L = N,N(1)-ethylene bis(salicylideneimine); H(2)L' = N,N(1)-bis(3-methoxysalicylidene)ethylene-1,2 diamine). The polymeric structures are formed by bridging neutral H(2)L units in the case of {[Ln(2)L(2)(CF(3)SO(3))(H(2)L)(4)(MeOH)] x CF(3)SO(3)}(n) (Ln = Eu (1), Nd (2) and Er (3)), and by acetate (OAc(-)) groups in [Yb(2)(L)(2)(OAc)(2)(MeOH)(2)](n) (4) and {[Tb(3)(L')(2)(OAc)(5)] x Et(2)O x (MeOH)(0.5)}(n) (5). The structures of 1-5 were determined by single crystal X ray crystallographic studies and the luminescence properties of 1 and 5 in MeOH solution were determined. PMID- 20023874 TI - Synthesis, structures and magnetic properties of polynuclear mixed-valence Mn(II)Mn(III) complexes containing 3-(2-phenol)-5-(pyridin-2-yl)-1,2,4-triazole ligand. AB - This paper reports the synthesis, crystal structures, and magnetic properties of a series of mixed-valence polynuclear manganese complexes bridged by a dianionic 1,2,4-triazole-based ligand L(2-) (H(2)L = 3-(2-phenol)-5-(pyridin-2-yl)-1,2,4 triazole). Complexes 1 to 3 show a similar binuclear structure, with the central core of [Mn(II)Mn(III)(L)(2)] and some other small molecules also coordinated to the two Mn centers. The two [Mn(II)Mn(III)(L)(2)] subunits can be further connected by different dicarboxylic acids to construct non-charged tetranuclear complexes 4 to 7. In these complexes, each deprotonated dianionic ligand L(2-) acts as a mu(2)-bridged ligand to coordinate two Mn centers via the 1,2,4 triazole ring. Bond valence sum (BVS) calculations and Mn surrounding bond lengths indicate that the Mn center coordinated to the pyridine N atoms in the +2 oxidation state, while another Mn center coordinated to the phenolic O atoms is in the +3 oxidation state. The Mn(III) centre shows Jahn-Teller elongation along one of the axes. Magnetic studies show the presence of ferromagnetic Mn(III) Mn(II) coupling in the binuclear (1-3) and tetranuclear (4-7) complexes with the magnetic coupling constant (J) ranging from 0.58 to 1.17 cm(-1), based on the Hamiltonian H = -2JS(1) x S(2) (S(1) = 5/2 and S(2) = 2). In all cases both z'J' (intermolecular interactions) and D parameter (Zero Field Splitting of the Mn (III) ion) has been introduced to improve the corresponding fit. The relationship of the coupling interaction between the Mn(II)/Mn(III) and their electronic structures is discussed. PMID- 20023875 TI - Transition metal complexes of 2-amino-3,5-dihalopyridines: Syntheses, structures and magnetic properties of (3,5-diCAPH)2CuX4 and (3,5-diBAPH)2CuX4. AB - A family of bis(2-amino-3,5-dihalopyridinium)tetrahalocuprate(II) compounds has been synthesized, including (3,5-diCAPH)2CuCl4 (1), (3,5-diCAPH)2CuBr4 (2), (3,5 diBAPH)2CuCl4 (3), and (3,5-diBAPH)2CuBr4 (4) [3,5-diCAPH = 2-amino-3,5 dichloropyridinium; 3,5-diBAPH = 2-amino-3,5-dibromopyridinium]. These complexes have been analyzed through single crystal X-ray diffraction and temperature dependent magnetic susceptibility. Compound 1 crystallizes in the P-1 space group and the tetrachlorocuprate ion is best described as possessing a distorted square planar geometry. Compounds 2-4 are structurally similar and crystallized in the P2(1)/n, P2(1)/c, and P2(1)/n space groups respectively. The tetrahalocuprate ions are best described as distorted tetrahedra. All four compounds show antiferromagnetic interactions and were fit to the uniform chain Heisenberg model with resulting 2J/kB values of -11.71(2) K, -2.21(1) K, -12.43 (2) K, and 1.36(1) K, respectively. The exchange values correlate well with the two-halide exchange pathway parameters. The unusual observation that the chloride complexes show stronger magnetic exchange than the bromide complexes provides strong support that the exchange can be strongly dependent upon the Cu-X...X angles and Cu-X...X-Cu torsion angles. PMID- 20023876 TI - Magnetically recyclable Ag-ferrite catalysts: general synthesis and support effects in the epoxidation of styrene. AB - A series of new magnetically recyclable catalysts (MRCs), Ag-M(1-x)Fe(2+x)O(4) (M = Co, Ni, Mn, Zn) nanocomposites, have been synthesized via a generalized one-pot route, and these materials prove to be efficient MRCs in the catalytic epoxidation of styrene. Characterization by X-ray diffraction (XRD), scanning electron microscopy (SEM), transmission electron microscopy (TEM), infrared (IR) and X-ray photoelectron spectroscopy (XPS), thermogravimetric analysis (TGA), and magnetic measurement reveal that the nanocomposites are assembled from well crystallized Ag and corresponding ferrite primary particles with the assistance of polyvinylpyrrolidone (PVP). The nanocomposite samples are comparable in Brunauer-Emmett-Teller (BET) specific surface areas and are superparamagnetic at room temperature. The as-prepared MRCs exhibit high activities in the epoxidation of styrene in toluene with tert-butylhydroperoxide (TBHP) as the oxidant and very high yields of styrene oxide can be achieved within 5 h of reaction. All the MRCs can be easily recovered by using a magnet and no decrease in the yield of styrene oxide has been observed after several consecutive rounds of reaction. It has been demonstrated that the ferrite supports affect the structure of Ag and the efficiency in oxygen supply and thus the catalytic performance. PMID- 20023877 TI - Phosphine-free synthesis of high quality ZnSe, ZnSe/ZnS, and Cu-, Mn-doped ZnSe nanocrystals. AB - High quality zinc blende ZnSe and ZnSe/ZnS core/shell nanocrystals have been synthesized by two converse injection methods (i.e. zinc precursor injection or selenium precursor injection) when Se-ODE complex was chosen as the phosphine free selenium precursor. Absorption spectroscopy, fluorescence spectroscopy, X ray diffraction (XRD), and transmission electron microscopy (TEM) were used to characterize the as-synthesized ZnSe and ZnSe/ZnS nanocrystals. The quality of the as-prepared ZnSe nanocrystals reached the same high level compared with the method using phosphine selenium precursors since the quantum yields were between 40 and 60% and photoluminescence (PL) full width at half-maximum (FWHM) was well controlled between 14 and 17 nm. The parameter window for the growth of high quality ZnSe nanocrystals was found to be much broader and monodisperse ZnSe nanocrystals were synthesized successfully even when the reaction temperature was set as low as 240 degrees C. As cores, such zinc blende ZnSe nanocrystals were also used to synthesize ZnSe/ZnS core/shell nanocrystals with high fluorescence quantum yields of 70%. Cu(2+) or Mn(2+) doped ZnSe nanocrystals were also synthesized by simply modifying this phosphine-free method. The emission range has been extended to 500 and 600 nm with the use of Cu(2+) and Mn(2+) dopants compared with the emission coverage of ZnSe at around 400 nm. This is the first totally "green approach" (i.e. phosphine-free synthesis) for the synthesis of high quality ZnSe, ZnSe/ZnS, and Cu(2+) or Mn(2+) doped ZnSe nanocrystals. PMID- 20023878 TI - Kinetics and mechanism of the Co(II)-assisted oxidation of L-ascorbic acid by dioxygen and nitrite in aqueous solution. AB - A detailed study of the oxidation of L-ascorbic acid by dioxygen and nitrite in water at pH 5.8 and 7.0, catalyzed by the octasulfophenyltetrapyrazinoporphyrazine complex of cobalt(II), was carried out using conventional spectrophotometric, low-temperature and high-pressure stopped flow techniques. The Co(II) complex activates L-ascorbic acid through an intramolecular one-electron oxidation step that involves the reduction of the octasulfophenyltetrapyrazinoporphyrazine. The reaction rate strongly depends on pH due to the different redox behaviour of the L-ascorbic acid/ascorbate species present in solution. Kinetic parameters for the different reaction steps of the catalytic process were determined. The final product of the reaction between L ascorbic acid and nitrite was found to be nitrous oxide. PMID- 20023879 TI - The influence of halogen substituents on the course of hydrogallation and hydroalumination reactions. AB - Treatment of trimethylsilylethynylbenzene derivatives with HGaCl(2) afforded products, [C(6)H(6-x){C(H)=C(SiMe(3))GaCl(2)}(x)], in which by a very fast cis/trans-rearrangement the Ga and H atoms occupied opposite sides of the resulting C=C double bonds. The stability of the cis-forms considerably increased upon application of 1,3-dibromo- and pentafluorophenylalkyne derivatives. Two pairs of cis/trans-isomers could be characterized by crystal structure determinations and allow the direct comparison of structural parameters. For the first time an equilibrium was detected between cis- and trans-forms in solution. Treatment of 1,4-di(tert-butylalkynyl)benzene with HAlR(2) (R = CMe(3), CH(2)CMe(3)) afforded cyclophane-type molecules by the release of AlR(3). Only the neopentyl derivative could be isolated and characterized by crystal structure determination. In contrast, the dibromo compound, 1,4-Br(2)-2,5-(Me(3)CC[triple bond]C)(2)C(6)H(2), yielded the simple addition product, C(6)H(2)Br(2){C(AlR(2))=C(H)CMe(3)}(2) (R = CMe(3)). Condensation was hindered in this case by intramolecular Al-Br interactions. Surprisingly, the simple addition product was also isolated from the reaction of 1,4-(Me(3)CC[triple bond]C)(2)C(6)H(4) with the relatively small hydride HAlEt(2). Solid-state NMR spectra of the product revealed strong intermolecular Al-C interactions involving the negatively charged terminal vinylic carbon atoms, to give one-dimensional coordination polymers. PMID- 20023880 TI - Synthesis, structural characterization and photoluminescence properties of rhenium(I) complexes based on bipyridine derivatives with carbazole moieties. AB - Three N,N-bidentate ligands, 5,5'-dibromo-2,2-bipyridine (L1) and two carbazole containing ligands of 5-bromo-5'-carbazolyl-2,2-bipyridine (L2), 5,5' dicarbazolyl-2,2'-bipyridine (L3), and their corresponding rhenium Re(CO)3Cl(L) complexes (ReL1-ReL3) have been successfully synthesized and characterized by elemental analysis, 1H NMR and IR spectra. Their photophysical properties and thermal analysis, along with the X-ray crystal structure analysis of L3 and complexes ReL1 and ReL3 are also described. In CH2Cl2 solution at room temperature, all complexes display intense absorption bands at ca. 220-350 nm, which can be assigned to spin-allowed intraligand (pi-->pi*) transitions, and the low energy broad bands in the 360-480 nm region are attributed to the metal to ligand charge-transfer d(Re)-->pi* (diimine) (MLCT). The introduction of carbazole moieties improves the MLCT absorption and molar extinction coefficient of these complexes. Upon excitation at the peak maxima, all complexes show strong emissions around 620 nm, which are assigned to d(Re)-->pi* (diimine) MLCT phosphorescence. The photoluminescence lifetime decay of Re(I) complexes were measured and the quantum efficiencies of the rhenium(I) complexes were calculated by using air-equilibrated [Ru(bpy)3]2+ x 2 Cl- aqueous solution as standard (phi(std) = 0.028). The complexes with appended carbazole moieties exhibit enhanced luminescence performances relative to ReL1. PMID- 20023881 TI - Coumarin-based luminescent ligand that forms helicates with dicationic metal ions. AB - The potentially hexadentate ligand L, which contains two terminal coumarin fluorophores, forms dinuclear double-stranded helicates with dicationic metal ions, giving species of the generic form [M(2)L(2)](4+). In solution the free ligand was fluorescent with emission attributed to the coumarin fluorophores (lambda(em) = 437 nm). The luminescent properties of the corresponding dimetallic helicates complexes were examined and revealed that the Zn(2+) complex demonstrates enhanced emission when compared to the parent ligand, whereas Co(2+), Cu(2+), Cd(2+) and Hg(2+) induce varying degrees of fluorescence quenching. In particular, comparative luminescence measurements at 77 K and room temperature showed that the quenching mechanism for [Cu(2)L(2)](4+) can be attributed to a photoinduced electron transfer. ESI-MS selectivity studies showed that in the presence of a mixture of metal dications no preference for any one metal ion was observed. PMID- 20023882 TI - A rearrangement of 1,1-linked four-membered rings to bicyclic bridgehead five membered rings: a phosphorus turn. AB - Novel rearrangements of 2,3,2',3'-tetra-tert-butyl-4,4'-bis-trimethylsilanyloxy 1,2,1',2'-tetrahydro-[1,1']biphosphetyl (7) to 1,2,5,6-tetra-tert-butyl-3,7-bis trimethylsilanyloxy-1H,5H-[1,2]diphospholo-[1,2-a][1,2]diphosphole (9) and of 3,4,3,4-tetra-tert-butyl-[1,1']biphosphetanyl-2,2'-dione (10) to 2,3,6,7-tetra tert-butyl-tetrahydro-[1,2]diphospholo[1,2-a][1,2]diphosphole-1,5-dione (12) were observed during sublimation at 150 degrees C in 0.1 mm vacuum. The C-P-C internal angles of the four-membered rings, which reflect steric strain in (7), were 71.9(2) degrees and 77.9(1) degrees; the C-P-P internal angles of the five membered rings of 9 were 90.88(6) degrees and 93.02(6) degrees after rearrangement. Reaction of 7 with metallic caesium provided the caesium salt of 3,4-di-tert-butyl-3,4-dihydro-phosphet-2-ol (15), which is the first example of a 2lambda-3-phospha-1-enolate being incorporated into four-membered ring. Due to its large size, the caesium cation has a rare coordination mode with both the phosphorus and oxygen atoms of the 2lambda-3-phospha-1-enolate, forming infinite polymeric chains in the solid state of 15. Two tert-butyl groups in the vicinal positions of compounds 7, 9, 10, 12 and 15 dictate the gauche conformations in compounds 7, 9, 10, 12 and 15, enforcing this stereoselectivity around the C-C-C portion of the rings in the enlargement reaction. PMID- 20023884 TI - Novel Re(I) dendrimers: synthesis, characterization and theoretical studies. AB - Four novel diimine rhenium(I) carbonyl complexes with the formula [Re(CO)3(L)Br], where L = 2-(4-(9H-carbazol-9-yl)phenyl)-1H-imidazo[4,5-f][1,10]phenanthroline (P1), 2-(4-(3,6-di-tert-butyl-9H-carbazol-9-yl)phenyl)-1H-imidazo-[4,5 f][1,10]phenanthroline (P2), 2-(4-(6-(9H-carbazol-9-yl)-9H-3,9-bicarbazol-9 yl)phenyl)-1H-imidazo[4,5-f][1,10]phenanthroline (D1), and 2-(4-(3',6'-di-tert butyl-6-(3,6-di-tert-butyl-9H-carbazol-9-yl)-9H-3,9-bicarbazol-9-yl)phenyl)-1H imidazo[4,5-f][1,10]phenanthroline (D2), have been successfully synthesized and fully characterized by 1H NMR, IR, and UV-Vis, etc. The luminescence quantum yields (LQYs) of the parent Re(I) complexes P1 and P2 are 0.13 and 0.16, respectively, which are much higher than the previously reported Re(I) dendrimers. The HOMOs and the LUMOs of P1 and P2 are calculated to be mainly composed of [d(Re) + pi(CO + Br)] and pi*(L) orbital, respectively. However, those of the Re(I) dendrimers D1 (LQY = 0.066) and D2 (LQY = 0.0048) are mainly localized on ligand L, indicating that the component of the metal-to-ligand charge-transfer d pi (Re) --> pi*(N-N) (MLCT) transitions in P1 and P2 should be more than those in D1 and D2. As a result, the higher LQYs of P1 and P2 are tentatively assigned to the disturbance of the MLCT transitions during the photoluminescence process. PMID- 20023883 TI - Palladium complexes of abnormal N-heterocyclic carbenes as precatalysts for the much preferred Cu-free and amine-free Sonogashira coupling in air in a mixed aqueous medium. AB - A series of new PEPPSI (Pyridine Enhanced Precatalyst Preparation Stabilization and Initiation) themed precatalysts of abnormal N-heterocyclic carbenes for the highly desirable Cu-free and amine-free Sonogashira coupling in air in a mixed aqueous medium is reported. Specifically, the PEPPSI themed (NHC)PdI2(pyridine) type precatalysts, 1b-4b, efficiently carried out the highly convenient Cu-free and amine-free Sonogashira coupling of aryl bromides and iodides with terminal acetylenes in air in a mixed aqueous medium. Complexes, 1b-4b, were synthesized by the direct reaction of the corresponding imidazo[1,2-a]pyridinium iodide salts, 1a-4a, with PdCl2 in pyridine in the presence of K2CO3 as a base while the imidazo[1,2-a]pyridinium iodide salts, 1a-4a, were in turn synthesized by the alkylation reactions of the respective imidazo[1,2-a]pyridine derivatives with alkyl iodides. The density functional theory (DFT) studies revealed that these imidazol-3-ylidene[1,2-a]pyridine derived abnormal carbenes are strongly sigma donating and consequently significantly weaken the catalytically important labile trans pyridine ligand in 1b-4b. PMID- 20023885 TI - 1,1,3,3-Tetramethylguanidine solvated lanthanide aryloxides: pre-catalysts for intramolecular hydroalkoxylation. AB - The synthesis and structural characterization of six 1,1,3,3-tetramethylguanidine (H-TMG) solvated lanthanide aryloxide complexes are reported. Ln[N{Si(CH3)3}2]3 (Ln = Nd, La) was reacted with two equivalents of both H-TMG and HOAr {HOAr = HOC6H2(CMe3)2-2,6 (H-DBP) or HOC6H2(CMe3)2-2,6-CH3-4 (H-4MeDBP)} and one equivelent of ethanol (HOEt) to yield the corresponding [Nd(H TMG)2(4MeDBP)2(OEt)] (1) and [La(H-TMG)2(DBP)2(OEt)] (2). Compounds 1 and 2 were further reacted with 4-pentyn-1-ol {HO(CH2)3C[triple bond]CH} to isolate [Nd(H TMG)2(4MeDBP)2{O(CH2)3C[triple bond]CH}] (3) and [La(H TMG)2(DBP)2{O(CH2)3C[triple bond]CH}] (4), respectively. Three equivalents of HOAr and one equivalent of H-TMG were additionally reacted with Ln[N{Si(CH3)3}2]3 to generate [Nd(4MeDBP)3(H-TMG)] (5) and [La(DBP)3(H-TMG)] (6). In order to examine the formation of 1-6, the interaction of H-TMG and HOAr was further examined in solution and the hydrogen bonded complexes (H-TMG:HOAr), 7 and 8, were isolated. Upon successful isolation of 1-6, the utility of 1, 2, 4 and 5 as pre-catalysts for the intramolecular hydroalkoxylation of 4-pentyn-1-ol was investigated. The bulk powders for all complexes were found to be in agreement with the crystal structures based on elemental analyses, FT-IR spectroscopy, and 1H and 13C NMR investigations. PMID- 20023886 TI - Templated assembly of mu(5)-CO3(2-) decanuclear praseodymium and neodymium clusters through spontaneous fixation of atmospheric carbon dioxide. AB - Reactions of Ln(III) acetate (Ln = Pr and Nd) and a polydentate Schiff-base in a mixture of methanol and acetonitrile resulted in the unprecedented assembly of novel Ln(10) aggregates containing two Ln(5) pentagons templated by mu(5)-CO(3)(2 ), introduced via spontaneous fixation of atmospheric carbon dioxide. Magnetic analysis using an expression including the ligand field effects and molecular field approximation indicates weak antiferromagnetic coupling between the metal ions. This synthetic approach may represent a promising new route toward the design of new lanthanide clusters and novel multifunctional materials. PMID- 20023887 TI - Remarkable thermal stability of Eu(4-phosphonobenzoate): structure investigations and luminescence properties. AB - A new 3D rare-earth hybrid material Eu(p-O(3)PC(6)H(4)COO) has been synthesised by a hydrothermal route from Eu(NO(3))(3) x 5 H(2)O and the rigid precursor, 4 phosphonobenzoic acid. The structure of Eu(p-O(3)PC(6)H(4)COO) has been solved by X-ray diffraction on a powder sample and is described as an inorganic network in which both carboxylic and phosphonic acid groups are linked to Eu ions forming a three-dimensional architecture. Thermal analysis performed on this compound has underlined its remarkable stability up to 510 degrees C and an optical study has been conducted to examine its luminescence properties that have been related to the structure of the material. The structural and luminescence properties have also been compared with the related material Eu phenylphosphonate. PMID- 20023890 TI - Dalton Transactions themed issue on metal anticancer compounds. PMID- 20023891 TI - In remembrance of Barnett Rosenberg. PMID- 20023893 TI - Metal complexes as photochemical nitric oxide precursors: potential applications in the treatment of tumors. AB - The bioregulatory molecule NO plays key roles in cancer biology and has been implicated in both tumor growth and suppression. Furthermore, it is a gamma radiation sensitizer that may enhance selective killing of neoplastic tissues. For these reasons, there is considerable interest in developing methods for NO delivery to specific physiological targets. In this Perspective, we describe ongoing investigations focused on photochemical methodologies to deliver therapeutic doses of NO to such targets utilizing transition metal complexes that are nitric oxide precursors. The photochemical strategy has the advantages that it allows for precise control of the timing, location, and dosage for the targeted delivery of a bioactive agent. PMID- 20023892 TI - Non-traditional platinum compounds for improved accumulation, oral bioavailability, and tumor targeting. AB - The five platinum anticancer compounds currently in clinical use conform to structure-activity relationships formulated (M. J. Cleare and J. D. Hoeschele, Bioinorg. Chem., 1973, 2, 187-210) shortly after the discovery that cis diamminedichloroplatinum(II), cisplatin, has antitumor activity in mice. These compounds are neutral platinum(II) species with two am(m)ine ligands or one bidentate chelating diamine and two additional ligands that can be replaced by water through aquation reactions. The resulting cations ultimately form bifunctional adducts on DNA. Information about the chemistry of these platinum compounds and correlations of their structures with anticancer activity have provided guidance for the design of novel anticancer drug candidates based on the proposed mechanisms of action. This article discusses advances in the synthesis and evaluation of such non-traditional platinum compounds, including cationic and tumor-targeting constructs. PMID- 20023894 TI - The Midas touch in cancer chemotherapy: from platinum- to gold-dithiocarbamato complexes. AB - The unquestionable therapeutic success of the anticancer drug cisplatin and its second- and third-generation analogues has triggered, in the past forty years, the development of several metal-based potential chemotherapeutic agents, most of which have failed to enter clinical trials. In this context, during the last decade, our research group has been making quite an effort to design a number of metal-dithiocarbamato derivatives that were expected, at least in principle, to resemble the main features of cisplatin together with higher activity, improved selectivity and bioavailability, and lower side-effects. Among all, gold(III) complexes have shown outstanding in vitro and in vivo antitumour properties and reduced or no systemic and renal toxicity, compared to the reference drug. Here, we summarize the results achieved to date, focusing on the mechanistic studies and the potential future developments opened up by our research work. PMID- 20023895 TI - The mechanism of action of platinum anticancer agents--what do we really know about it? AB - Although 40 years have passed since the discovery of the anticancer activity of cisplatin, the mechanism of action of the drug is unclear. There are several working hypotheses that guide the researchers in this field. Unexpected results that we obtained cannot be reconciled with some of those assumptions. Our main intention is to call upon researchers in the field to re-examine the working hypotheses that may have been formulated on the basis of outdated methodologies. PMID- 20023897 TI - Novel and emerging approaches for the delivery of metallo-drugs. AB - Cisplatin and its derivatives are the most widely used clinical anticancer agents. They bring enormous benefits to patients but are also associated with unpleasant side-effects because of their abilities to interact with biomolecules other than the target DNA and their broad tissue toxicity across the body. While two molecular re-designs of cisplatin have entered worldwide clinical use (carboplatin and oxaliplatin) and many more have been trialled, these side effects and drawbacks remain. Recently new strategies have been developed to attempt to decrease these side effects and/or modify the tissue activity spectrum through more localized and effective delivery of the drug to the desired targets. In this review we present an overview of the principal approaches that have been explored, ranging from conjugation to biomolecular vectors or polymers, through pro-drug strategies, to adsorption on ceramic materials and encapsulation in macrocycles, nanotubes and nanocapsules, biomolecules and polymers. PMID- 20023896 TI - Photoactivated chemotherapy (PACT): the potential of excited-state d-block metals in medicine. AB - The fields of phototherapy and of inorganic chemotherapy both have long histories. Inorganic photoactivated chemotherapy (PACT) offers both temporal and spatial control over drug activation and has remarkable potential for the treatment of cancer. Following photoexcitation, a number of different decay pathways (both photophysical and photochemical) are available to a metal complex. These pathways can result in radiative energy release, loss of ligands or transfer of energy to another species, such as triplet oxygen. We discuss the features which need to be considered when developing a metal-based anticancer drug, and the common mechanisms by which the current complexes are believed to operate. We then provide a comprehensive overview of PACT developments for complexes of the different d-block metals for the treatment of cancer, detailing the more established areas concerning Ti, V, Cr, Mn, Re, Fe, Ru, Os, Co, Rh, Pt, and Cu and also highlighting areas where there is potential for greater exploration. Nanoparticles (Ag, Au) and quantum dots (Cd) are also discussed for their photothermal destructive potential. We also discuss the potential held in particular by mixed-metal systems and Ru complexes. PMID- 20023898 TI - Dual anti-angiogenic and cytotoxic properties of ruthenium(III) complexes containing pyrazolato and/or pyrazole ligands. AB - An oxo-bridged diruthenium(III) complex containing pyrazolato and pyrazole ligands is stable against ascorbic-acid reduction, induces apoptosis (60%, 48 h) against HeLa cells at 10 microM level and exhibits promising anti-angiogenic activity at its sub-cytotoxic concentrations. Other mononuclear ruthenium(III) complexes containing pyrazole ligands [Ru(pz)(4)X(2)](+) exhibit dual anti angiogenic and cytotoxic properties. PMID- 20023899 TI - Interactions of ruthenium coordination cubes with DNA. AB - The interactions of four octacationic ruthenium coordination cubes with duplex and quadruplex (telomeric and c-myc) DNA have been studied by FID and SPR, showing these cubes to bind strongly to tetrastranded DNA structures. PMID- 20023900 TI - Synthesis, characterization, in vitro antitumoral investigations and interaction with plasmid pBR322 DNA of R2eddp-platinum(IV) complexes (R = Et, n-Pr). AB - The studies on synthetic, spectroscopic and biological properties of platinum(IV) complexes, [PtCl(4)(R(2)eddp)] (R = Et, 1; n-Pr, 2), containing kappa(2)N,N' bidentate ligands, esters of ethylenediamine-N,N'-di-3-propionic acid (HOOCCH(2)CH(2)NHCH(2)CH(2)NHCH(2)CH(2)COOH, H(2)eddp), are reported. Complexes have been characterized by infrared, (1)H and (13)C NMR spectroscopy and elemental analysis and it was concluded that the coordination of the ligands occurs via nitrogen donor atoms of the ester ligands (R(2)eddp). Cytotoxicity studies were performed for ligand precursors and corresponding platinum(IV) complexes. Although the n-Pr(2)eddp.2HCl itself showed no activity (IC(50) values > 125 microM) in selected cell lines, the activity of complex 2, via apoptotic mode of cell death, has increased significantly for a broad range of cancer cell lines tested in vitro (IC(50) = 8.6-49 microM). As it was found that complexes 1 and 2 are able to interact with pBR322 plasmid DNA, platinum(IV) complexes of this type may act as drugs and pro-drugs. PMID- 20023901 TI - A novel ruthenium(III) complex with a tridentate dianionic P,O,O-ligand showing high cytotoxic activity. AB - Tris(o-anisyl)phosphane was reacted with ruthenium(III) chloride forming a novel Ru(III) complex of formula [Ru(PAn(3))(P(An)(phenolate)(2))Cl] containing a tridentate dianionic P,O,O-ligand and a bidentate neutral P,O ligand. The tridentate ligand was formed by elimination of two methyl groups from the starting ligand in the reaction with the ruthenium salt. The molecular structure was determined by single crystal X-ray diffraction analysis. The compound shows a high cytotoxic activity in ovarian cancer cell lines comparable with cisplatin and overcoming cisplatin-resistance. PMID- 20023902 TI - Excellent correlation between cathepsin B inhibition and cytotoxicity for a series of palladacycles. AB - The reaction of the five- or six-membered C,N or C,S-palladacycles [(L)PdCl](2) with PTA (1,3,5-triaza-7-phosphaadamantane) led to the monomeric complexes [(L)Pd(PTA)Cl] 6a, 6b and 7 where LH= N,N-dimethyl-1-phenylmethanamine, benzyl(methyl)sulfane or 1-methyl-5-phenyl-1H-benzo[e][1,4]diazepin-2(3H)-one respectively. Dimeric complexes have also been synthesised: [Pd(2)L(2)(mu dppe)Cl(2)], where LH = 1-methyl-5-phenyl-1H-benzo[e][1,4]diazepin-2(3H)-one (1a), (R)- or (S)-3-isopropyl-1-methyl-5-phenyl-1H-benzo[e][1,4]diazepin-2(3H) one (1b, 1c), [Pd(2)L(2)(mu-dppf)Cl(2)], where L= 1-methyl-5-phenyl-1H benzo[e][1,4]diazepin-2(3H)-one (4a) or (R)-3-isopropyl-1-methyl-5-phenyl-1H benzo[e][1,4]diazepin-2(3H)-one (4b), respectively, and dppe = 1,2 bis(diphenylphosphino)ethane, dppf = 1,1'-bis(diphenylphosphino)ferrocene. The complexes were characterised in solution, by (1)H and (31)P NMR spectroscopy, and single crystals of complexes 6b and 7 were studied in the solid state by X-ray crystallography. The palladacycles were evaluated for in vitro activity as cytotoxic agents on A2780/S cells and also as cathepsin B inhibitors, an enzyme implicated in a number of cancer related events. PMID- 20023903 TI - Mono- and dinuclear platinum(II) compounds with 5,7-dimethyl-1,2,4-triazolo[1,5 a]pyrimidine. Structure, cytotoxic activity and reaction with 5'-GMP. AB - Mono- and dinuclear platinum(II) coordination compounds of formula cis [PtCl(2)(NH(3))(dmtp)], 1, cis-[PtCl(2)(dmtp)(2)], 2 and {H(+)[C(28)H(32)Cl(2)N(16)Pt(2)](2+)(NO(3))(3)(H(2)O)(6)}, 3, in which dmtp is 5,7-dimethyl-1,2,4-triazolo[1,5-a]pyrimidine have been synthesized and characterized by infrared and by (1)H, (13)C, (195)Pt NMR spectroscopy. The coordination units of the cationic species of formula [Pt(2)(mu dmtp)(2)Cl(2)(dmtp)(2)](2+) are built up by two platinum atoms in a square-planar environment. Two sites are occupied by two 5,7-dimethyl-1,2,4-triazolo[1,5 a]pyrimidine (dmtp) bridging ligands which are linked to both metal atoms through their nitrogen atoms in positions 3 and 4. The other two positions are occupied by one monodentate 5,7-dimethyl-1,2,4-triazolo[1,5-a]pyrimidine (dmtp) molecule and a coordinated chloride. This compound is the first in which the same triazolopyrimidine ligand (dmtp) coordinates to a metal ion in two different ways, i.e. bridging bidentate and non-bridging monodentate. In addition, the interaction of compounds 1 and 2 with 5'-GMP was investigated in solution by (1)H NMR spectroscopy. The cytotoxicity of all the new platinum(II) compounds were studied by using two different cell lines: T47D (breast cancer) and HCV29T (bladder cancer). PMID- 20023904 TI - Synthetic strategies towards ruthenium-porphyrin conjugates for anticancer activity. AB - The conjugation of porphyrins to metal fragments is a strategy for making new compounds that are expected to combine the phototoxicity and the tumour localization properties of the porphyrin chromophore with the cytotoxicity of the metal fragment for additive antitumour effect. We report here the preparation of new classes of porphyrin-ruthenium conjugates with potential bio-medical applications. Ruthenium was chosen because several Ru compounds have shown promising anticancer activity. The conjugation with the porphyrin moiety was accomplished either through peripheral pyridyl rings (e.g.meso-4' tetrapyridylporphyrin, 4'TPyP) or through bpy units (e.g.meso-(p-bpy phenyl)porphyrins, bpy(n)-PPs, n = 1-4). The number of Ru fragments attached to the porphyrins ranges from 1 to 4 and the total charge of the conjugates from -4 to +8. Different types of peripheral fragments, both Ru(III) and Ru(II), have been used: in some cases they are structurally similar to established anticancer compounds. Examples are [Na](4)[4'TPyP{trans-RuCl(4)(dmso-S)}(4)] (2), that bears four NAMI-type Ru(III) fragments, or [4'TPyP{Ru([9]aneS3)(en)}(4)][CF(3)SO(3)](8) (3) and [bpy(4)-PP{Ru([9]aneS3)(dmso-S)}(4)][CF(3)SO(3)](8) (9) (en = ethane-1,2 diamine, [9]aneS3 = 1,4,7-trithiacyclononane) that have four half-sandwich Ru(II) compounds. The Ru fragments may either contain one or more labile ligands, such as in 2 or in 9, or be coordinatively saturated and substitutionally inert, such as in 3 or in [bpy(4)-PP{Ru([12]aneS4)}(4)][CF(3)SO(3)](8) (11) ([12]aneS4 = 1,4,7,10-tetrathiacyclododecane). Most of the ruthenium-porphyrin conjugates described in this work are soluble--at least moderately--in aqueous solution and are thus suitable for biological investigations, in particular for cytotoxicity and photo-cytotoxicity tests. PMID- 20023905 TI - Microwave synthesis of mixed ligand diimine-thiosemicarbazone complexes of ruthenium(II): biophysical reactivity and cytotoxicity. AB - A novel microwave-assisted synthetic method has been used to synthesise a series of mixed ligand ruthenium(II) compounds containing diimine as well as bidentate thiosemicarbazone ligands. The compounds contain the diimine 1,10-phenanthroline (phen) or 2,2'-bipyridine (bpy) and the thiosemicarbazone is derived from 9 anthraldehyde. Based on elemental analyses and spectroscopic data, the compounds are best formulated as [(phen)(2)Ru(thiosemicarbazone)](PF(6))(2) and [(phen)(2)Ru(thiosemicarbazone)](PF(6))(2) where thiosemicarbazone = 9 anthraldehydethiosemicarbazone, 9-anthraldehyde-N(4)-methylthiosemicarbazone, and 9-anthraldehyde-N(4)-ethylthiosemicarbazone. Fluorescence competition studies with ethidium bromide, along with viscometric measurements suggests that the complexes bind calf thymus DNA (CTDNA) relatively strongly via an intercalative mode possibly involving the aromatic rings of the diimine ligands. The complexes show good cytotoxic profiles against MCF-7 and MDA-MB-231 (breast adenocarcinoma) as well as HCT 116 and HT-29 (colorectal carcinoma) cell lines. PMID- 20023906 TI - Cytotoxicity, cellular localisation and biomolecular interaction of non-covalent metallo-intercalators with appended sex hormone steroid vectors. AB - A range of terpyridine platinum(II) metallo-intercalators with bioactive steroids attached has been created with the aim of localizing cytotoxic drugs. Complexes where the steroid does not interfere with access to the terpyridine are shown to retain potent cytotoxicity and show certain selectivity towards their natural receptors. Because the intercalation of the terpyridine moiety between the bases of the DNA is the origin of the biological activity, a dramatic decrease of the activity is observed when the access to the terpyridine unit is hindered by the steroidal unit. PMID- 20023907 TI - On the many roles of NH3 ligands in mono- and multinuclear complexes of platinum. AB - The role of the NH(3) ligands in the highly successful antitumour agents cisplatin and carboplatin is not fully understood. Suggestions that the ammonia ligands are involved in target recognition through hydrogen bond formation, e.g. with guanine-O6, have been questioned. Here, we review the roles and functions of NH(3) ligands of cis-PtCl(2)(NH(3))(2) and likewise of its trans-isomer in complexes with model nucleobases as well as other N-heterocyclic ligands. Specifically, their roles in hydrogen bonding interactions with nucleobases as well as anions, the influence on acid-base properties of co-ligands, their involvement in condensation reactions, as well as a variety of displacement reactions will be examined. As a result, it can be stated that the ammonia ligands in cis- and trans-Pt(II)(NH(3))(2) entities display additional features to those generally discussed in the last four decades since the discovery of the antitumour activity of cisplatin. PMID- 20023908 TI - Iron(III) complexes of fluorescent hydroxamate ligands: preparation, properties, and cellular processing. AB - Iron(III) complexes containing hydroxamic acid fluorophores were investigated as models of hypoxia selective prodrugs in vitro. Two complexes were synthesised, [Fe(c343haH)(3)] and [Fe(salen)(c343haH)]. The fluorescence of the hydroxamate coumarin fluorophore was almost completely quenched on coordination to the iron(III) centre in [Fe(c343haH)(3)]. However, quenching was minimal for [Fe(c343haH)(salen)] in aqueous media and we propose that the fluorescence results from structural rearrangements that occur because of the inherent strain in the iron-salen structure. Fluorescence was also measured in the presence of the cellular reductants ascorbic acid and cysteine. Fluorescence intensity increased over time, with the most rapid return of fluorescence occurring over a two hr period. The rapid fluorescence return indicates that the complexes undergo ligand release, either via reduction followed by aquation, or via direct ligand exchange with the reductants. Electrochemical studies demonstrated that both complexes have very negative reduction potentials. Furthermore, [Fe(c343haH)(salen)] was shown to exhibit quasi-reversibility of reduction. The distribution of the free hydroxamate ligand and the complexes were monitored in A2780 cells. The free ligand displayed non-specific distribution, which differed from the nucleolar distribution of [Fe(c343haH)(3)] and the lysosomal accumulation of [Fe(c343haH)(salen)] over time. Thus the results of the present study show that iron(III) complexes present a viable model for monitoring hydroxamate fluorophore displacement in vitro to determine the fate of prodrugs. PMID- 20023909 TI - Naphthalimide gold(I) phosphine complexes as anticancer metallodrugs. AB - Gold(I) phosphine complexes exhibit promising properties for anticancer drug development. Here we report on a series of gold(I) phosphine complexes containing a naphthalimide ligand. Strong antiproliferative effects were observed in MCF-7 breast cancer cells as well as in HT-29 colon carcinoma cells. The cellular and nuclear gold levels were increased compared to analogues, in which the naphthalimide ligand was replaced by a chloro ligand. Compound 4a was selected for more detailed biochemical and biological studies, which revealed solvent dependent fluorescence emission, uptake of the compound into the organelles of tumor cells as well as antiangiogenic effects concerning angiogenesis and tumor induced angiogenesis in vivo. Antiangiogenic properties of 4a were observed in two different zebrafish angiogenesis models, including a tumor-cell induced neovascularization assay. PMID- 20023910 TI - Anticancer activity of heteroleptic diimine complexes of dirhodium: a study of intercalating properties, hydrophobicity and in cellulo activity. AB - The series of complexes cis-[Rh(2)(mu-O(2)CCH(3))(2)(dppn)(L)](2+), where dppn = benzo[i]dipyrido[3,2-a:2',3'-c] phenazine, and L = bpy (2,2'-bipyridine) (1), phen (1,10-phenanthroline) (2), dpq (dipyrido[3,2-f:2',3'-h]quinoxaline) (3), dppz (dipyrido[3,2-a:2',3'-c]phenazine) (4), and dppn (5) were synthesized and their effect on the human cancer cells HeLa and COLO-316 was monitored. Complexes 1 and 2 interact with DNA through intercalation, whereas compounds 3-5 bind only electrostatically. It was found that the dirhodium complex 4 is the most effective compound at inhibiting cell viability of the human cancer cells HeLa and COLO-316. A general conclusion is that the hydrophobicity of the compounds correlates with their in cellulo activity in both cell lines. The ability of the compounds to reach nuclear DNA and form adducts was explored using the comet assay. The results indicate that compounds 1-5 either do not form adducts with DNA that are detrimental to the cell or that they are successfully repaired by the cellular machinery. The results of an annexin V assay indicate that compounds 1-4 trigger apoptosis, whereas compound 5 clearly does not. These findings are significant because they support the contention that dirhodium complexes can be tuned to direct their effect to cellular targets other than nuclear DNA. PMID- 20023911 TI - Divalent later transition metal complexes of the traditional chinese medicine (TCM) liriodenine: coordination chemistry, cytotoxicity and DNA binding studies. AB - Liriodenine (L), a natural alkaloid, was isolated as an active component from the anticancer traditional Chinese medicine (TCM), Zanthoxylum nitidum. It reacted with Mn(II), Fe(II), Co(II) and Zn(II) to afford four metal complexes: [MnCl(2)(L)(2)] (1), [FeCl(2)(L)(2)] (2), [Co(L)(2)(H(2)O)(2).Co(L)(2)(CH(3)CH(2)OH)(2)](ClO(4))(4) (3), and [Zn(2)(L)(2)(mu(2)-Cl)(2)Cl(2)] (4), which were characterized by elemental analysis, IR, ESI-MS. Their crystal structures were determined by the single crystal X-ray diffraction method. The in vitro cytotoxicity of L and complexes 1 4 against 10 human tumour cell lines was assayed. Some of these metal-based compounds exhibited enhanced cytotoxicity vs. free L to selected tumour cell lines. The binding properties of L and its complexes 1-4 to ct-DNA were investigated by spectroscopic methods and viscosity measurements. Agarose gel electrophoresis experiments were also carried out to evaluate their unwinding ability towards plasmid DNA and their inhibition towards Topoisomerase I. All the results indicate that complexes 1-4 may bind more intensively to the DNA helix than does L, and intercalative binding for complexes 1-4 and electrostatic interactions for complexes 3-4 to DNA should be considered. For complex 4, covalent binding to DNA may exist. Of special note, all these metal complexes effectively inhibit Topoisomerase I even at low concentration (< or = 10 microM). PMID- 20023912 TI - Cytotoxicity of the traditional chinese medicine (TCM) plumbagin in its copper chemistry. AB - The anticancer traditional Chinese medicine (TCM), plumbagin (PLN), was isolated from Plumbago Zeylanica. Reaction of plumbagin with Cu(II) salt, afforded [Cu(PLN)(2)].2H(2)O (1). With 2,2'-bipyridine (bipy) as a co-ligand, PLN reacts with Cu(II) to give [Cu(PLN)(bipy)(H(2)O)](2)(NO(3))(2).4H(2)O (2). 1 and 2 were characterized by elemental analysis, IR, ESI-MS spectra. Their crystal structures were determined by single crystal X-ray diffraction methods. The in vitro cytotoxicity of PLN, 1 and 2 against seven human tumour cell lines was assayed. The metal-based compounds exhibit enhanced cytotoxicity vs. that of free PLN, suggesting that these compounds display synergy in the combination of metal ions with PLN. The binding properties of PLN, 1 and 2 to DNA were investigated through UV-vis, fluorescence, CD spectra, and gel mobility shift assay, which indicated that 1 and 2 were non-covalent binding and mainly intercalated the neighboring base pairs of DNA. PLN, 1 and 2 exhibit inhibition activity to topoisomerase I (TOPO I), but 1 and 2 were more effective than PLN. PMID- 20023913 TI - Gadolinium texaphyrin (Gd-Tex)-malonato-platinum conjugates: synthesis and comparison with carboplatin in normal and Pt-resistant cell lines. AB - The synthesis of a new PEG-solubilized gadolinium texaphyrin (Gd-Tex) conjugate containing a malonate-Pt(NH(3))(2) moiety is described. The effect of the tumor localizing Gd-Tex macrocycle on platinum activity was evaluated in cell culture. The malonate moiety, analogous to that present in carboplatin, is expected to release an aquated Pt(NH(3))(2) species under physiological conditions. The half life in phosphate-buffered saline was found to be ca. 3 days at room temperature, and the hydrolytic product released from the conjugate was collected and confirmed as Pt-based by flameless atomic absorption spectrophotometry. Anti proliferative activity was tested using A549 human lung cancer and A2780 human ovarian cancer cell lines. In both cell lines, the activity of the Gd-Tex conjugate was found to be similar to that of carboplatin. Efficacy against a Pt resistant ovarian cell line greater than that displayed by carboplatin was also observed. PMID- 20023914 TI - Synthesis and biological studies of some gold(I) complexes containing functionalised alkynes. AB - The propargyl ethers 7-chloro-(4-propargyloxy)quinoline, 1 propargyloxynaphthalene and 2-propargyloxybenzophenone react with [AuCl(PPh(3))] in the presence of KOH to give the gold(I) alkynyl complexes [Au(C[triple bond]COCH(2)Ar)(PPh(3))] in good yields. The compounds were fully characterised by spectroscopic methods and were subsequently examined for their biological activity against four tumour cell lines as well as their activity against Plasmodium falciparum, the parasite responsible for malaria. The compounds show antiproliferative effects in human cancer cells with IC(50) values ranging from 0.4-12 microM. PMID- 20023915 TI - DNA cleavage and antitumour activity of platinum(II) and copper(II) compounds derived from 4-methyl-2-N-(2-pyridylmethyl)aminophenol: spectroscopic, electrochemical and biological investigation. AB - The reaction of the redox-active ligand, Hpyramol (4-methyl-2-N-(2 pyridylmethyl)aminophenol) with K(2)PtCl(4) yields monofunctional square-planar [Pt(pyrimol)Cl], PtL-Cl, which was structurally characterised by single-crystal X ray diffraction and NMR spectroscopy. This compound unexpectedly cleaves supercoiled double-stranded DNA stoichiometrically and oxidatively, in a non specific manner without any external reductant added, under physiological conditions. Spectro-electrochemical investigations of PtL-Cl were carried out in comparison with the analogue CuL-Cl as a reference compound. The results support a phenolate oxidation, generating a phenoxyl radical responsible for the ligand based DNA cleavage property of the title compounds. Time-dependent in vitro cytotoxicity assays were performed with both PtL-Cl and CuL-Cl in various cancer cell lines. The compound CuL-Cl overcomes cisplatin-resistance in ovarian carcinoma and mouse leukaemia cell lines, with additional activity in some other cells. The platinum analogue, PtL-Cl also inhibits cell-proliferation selectively. Additionally, cellular-uptake studies performed for both compounds in ovarian carcinoma cell lines showed that significant amounts of Pt and Cu were accumulated in the A2780 and A2780R cancer cells. The conformational and structural changes induced by PtL-Cl and CuL-Cl on calf thymus DNA and phiX174 supercoiled phage DNA at ambient conditions were followed by electrophoretic mobility assay and circular dichroism spectroscopy. The compounds induce extensive DNA degradation and unwinding, along with formation of a monoadduct at the DNA minor groove. Thus, hybrid effects of metal-centre variation, multiple DNA-binding modes and ligand-based redox activity towards cancer cell-growth inhibition have been demonstrated. Finally, reactions of PtL-Cl with DNA model bases (9-Ethylguanine and 5'-GMP) followed by NMR and MS showed slow binding at Guanine-N7 and for the double stranded self complimentary oligonucleotide d(GTCGAC)(2) in the minor groove. PMID- 20023916 TI - 1 : 2 Adducts of copper(I) halides with 1,2-bis(di-2-pyridylphosphino)ethane: solid state and solution structural studies and antitumour activity. AB - The 1 : 2 adducts of copper(I) halides with 1,2-bis(2-pyridylphosphino)ethane (d2pype) have been synthesized and solution properties characterized by variable temperature (1)H, (31)P and (65)Cu NMR spectroscopy. Single-crystal structure determinations for the chloride, bromide and iodide complexes show these to crystallize from acetonitrile in the triclinic space group P1 as isostructural centrosymmetric dimers [(d2pype)Cu(mu-d2pype)(2)Cu(d2pype)]X(2).(solvent) with a approximately 12.6, b approximately 12.7, c approximately 15.3 A, alpha approximately 84, beta approximately 67, gamma approximately 84 degrees. In contrast to the analogous AuCl:2(d2pype) and AgNO(3):2(d2pype) adducts, in solution these CuX:2(d2pype) adducts (where X = Cl, Br and I) exist almost exclusively as bis-chelated monomeric [Cu(d2pype)(2)]X; evidence for an equilibrium between monomeric and dimeric forms is detected only for the CuCl adduct in methanol. Cytotoxicity studies in two human breast cancer lines and two matched liver progenitor cell lines indicate that [Cu(d2pype)(2)]Cl is non selectively toxic to both non-tumourigenic and tumourigenic cells. However, the analogous Au(I) compound [Au(d2pype)(2)]Cl, is toxic to highly tumourigenic cells and more selective in its toxicity to tumourigenic cells compared to non tumourigenic cells. The significance of these results to the further development of selective, mitochondria-targeted, Au(I) antitumour complexes is discussed. PMID- 20023917 TI - Synthesis, oxidation chemistry and cytotoxicity studies on ferrocene derivatives of diethylstilbestrol. AB - A series of compounds is described in which one of the ethyl groups in diethylstilbestrol has been replaced by a ferrocenyl substituent. Only those derivatives incorporating phenol moieties underwent isomerisation from the Z to the E form, and some of them could be chemically oxidized to a quinone species. The compounds were less cytotoxic against hormone-independent MDA-MB-231 breast cancer cell lines than their corresponding ferrocenyl phenyl or phenol isomers in which the ferrocene and ethyl moieties are linked to the same carbon atom. The biochemical results were evaluated in conjunction with information obtained from electrochemical and chemical oxidation experiments. PMID- 20023918 TI - Inert ruthenium half-sandwich complexes with anticancer activity. AB - In this study, we investigate the anticancer properties of an inert half-sandwich metal complex scaffold. UV melting experiments with duplex DNA and (1)H-NMR experiments with 9-ethylguanine reveal that the apoptotic ruthenium complex DW12 does not interact with DNA. On the other hand, diminishing the kinase inhibition properties of DW12 by methylating the maleimide nitrogen (DW12-Me) abolishes the anticancer activity. Furthermore, the incorporation of a fluorine into the pyridine moiety (NP309) improves the IC(50) value for glycogen synthase kinase 3 (GSK-3) and at the same time the cytotoxicity, implying that the anticancer activity correlates with the inhibition of GSK-3 and maybe other not yet identified kinases. We demonstrate in Burkitt-like lymphoma (BJAB) cells that NP309 is not necrotic but induces apoptosis and that this apoptosis is mediated by a loss of the mitochondrial membrane potential, caspase-9 processing, and is partly dependent on Bcl-2 expression. In addition, NP309 efficiently induces apoptosis in vincristine- and cytarabine-resistant human B-cell precursor cell lines. PMID- 20023919 TI - The role of bridging ligands in determining DNA-binding ability and cross-linking patterns of dinuclear platinum(II) antitumour complexes. AB - The DNA binding ability and binding mode of platinum complexes are crucial factors that govern their cytotoxic activity. In this work, circular dichroism spectroscopy, gel electrophoresis and MALDI-TOF MS spectrometry combined with enzymatic degradation have been used to elucidate the role of bridging ligands in DNA-binding ability and cross-linking patterns of two dinuclear antitumour active platinum(II) complexes, {[cis-Pt(NH(3))(2)Cl](2)L1}(NO(3))(2) (1, L1= 4,4' methylenedianiline) and {[cis-Pt(NH(3))(2)Cl](2)L2}(NO(3))(2) (2, L2 = alpha,alpha'-diamino-p-xylene). Although both complexes have two cis-diammine Pt(II) moieties (1,1/c,c), complex 1 exhibits much higher DNA-binding ability than complex 2. The former readily forms both 1,3- and 1,4-intrastrand cross links with DNA oligonucleotides, while the latter preferentially forms 1,4- rather than 1,3-intrastrand cross-links. Cytotoxicity studies against a human non small-cell lung cancer cell line (A549) demonstrate that complex 1 has higher activity than 2. These results show that the linker properties play a critical role in controlling the DNA-binding and cross-linking abilities and in modulating the cytotoxicity of dinuclear platinum complexes. PMID- 20023920 TI - Reactions of palladium and gold complexes with zinc-thiolate chelates using electrospray mass spectrometry and X-ray diffraction: molecular identification of [Pd(bme-dach)], [Au(bme-dach]+ and [ZnCl(bme-dach)]2Pd. AB - The reaction between the complexes [MCl(L)]Cl(x) (L = 2,2',2''-terpyridine, terpy and dien, diethylenetriamine; M = Pd, x = 1; M = Au, x = 2) and [Zn(bme dach)](2), an N(2)S(2)-Zn-thiolate bridged dimer used to mimic zinc finger protein sites, was studied by Electrospray Ionisation Mass Spectrometry and the structures of some of the products confirmed by X-ray crystallography. All reactions investigated in this work gave heteronuclear (Zn-thiolate)-metal products, the predominant species being the trinuclear dithiolate-bridged aggregate {[Zn(bme-dach)](2)M}(n+) (M = Pd, Au). X-Ray diffraction studies verified the molecular structure of [{ZnCl(bme-dach)}(2)Pd], and further confirmed that the zinc within the [Zn(bme-dach)](2) unit was retained within the N(2)S(2) binding site. The Zn-bound thiolates form stable thiolate bridges to Pd(2+) in a stair-step shape, held together by a planar PdS(4) center. In addition, both zinc atoms maintained penta-coordinate coordination with apical chloride ligands rather than the more commonly observed tetrahedral geometry. Further, [Pd(bme-dach)] was directly synthesized for X-ray structural characterization of the metal exchanged product observed in mass spectrometry experiments. In the case of Au compounds, the reactions were very fast and the products were similar for both [AuCl(L)]Cl(2) (L = terpy and dien) starting materials. In addition to the multimetallic Zn,Au,Zn aggregate formation, the predominant species from the reaction between [Zn(bme-dach)](2) and both Au compounds was the [Au(bme-dach](+) cation observable via ESI-MS, suggesting Zn/Au metal exchange immediately after mixing the compounds. The direct synthesis of [Au(bme-dach)]BPh(4) confirmed the molecular structure of this species through X ray crystallography. The reactivity profile of Pd(2+) and Au(3+) species is compared with previous studies using the isostructural Pt compounds and the biological relevance of the results discussed. PMID- 20023921 TI - Synthesis, characterization, and cytotoxicity of dinuclear platinum bisphosphonate complexes to be used as prodrugs in the local treatment of bone tumours. AB - For over 30 years cisplatin has been one of the most active antitumour agents in clinical use, nevertheless research for overcoming cisplatin toxicity and resistance or for improving its efficacy has never ceased. In this context we have recently proposed dinuclear Pt complexes with bridging geminal bisphosphonates as novel Pt-prodrugs with potential activity at the bone surface after embedment in inorganic matrices and implantation at the tumour site. In the present paper we report the synthesis and full characterization of four new platinum complexes having a dinuclear structure with a bisphosphonate (2-ammonium 1-hydroxyethane-1,1-diyl-bisphosphonate or 3-ammonium-1-hydroxypropane-1,1-diyl bisphosphonate, AHBP-H and PAM-H, respectively) acting as a bridging ligand between two platinum moieties (cis-[Pt(NH(3))(2)](2+), directly related to cisplatin, and [Pt(cis-1,4-DACH)](2+), known to be able to overcome the cisplatin resistance). Moreover, as a preliminary investigation, the in vitro cytotoxicity of the new complexes has been evaluated on a panel of 13 human tumour cell lines including cisplatin- and multidrug-resistant sublines. PMID- 20023922 TI - Synthesis, molecular structure and evaluation of new organometallic ruthenium anticancer agents. AB - A number of new ruthenium compounds have been synthesised, isolated and characterised, which exhibit excellent cytotoxicity against a number of different human tumour cell lines including a defined cisplatin resistant cell line and colon cancer cell lines. Addition of hydrophobic groups to the ruthenium molecules has a positive effect on the cytotoxicity values. Evidence is provided that, after incubation of a ruthenium compound with a 46 mer oligonucleotide duplex and subsequent nuclease treatment, ruthenium is bound to a guanine residue. PMID- 20023923 TI - Nuclear targets of photodynamic tridentate ruthenium complexes. AB - Octahedral ruthenium complexes, capable of photodynamic singlet oxygen production at near 100% efficiency, were shown to cause light-dependent covalent crosslinking of p53 and PCNA subunits in mammalian cells and cell lysates. Azide, a singlet oxygen quencher, greatly reduced the p53 photocrosslinking, consistent with the idea that singlet oxygen is the reactive oxygen species involved in p53 photocrosslinking. A photodynamically inactive ruthenium complex, [Ru(tpy)(2)](2+) (tpy = [2,2';6',2'']-terpyridine), had no effect on p53 or PCNA photocrosslinking. Photodynamic damage to p53 has particular relevance since p53 status is an important determinant of phototoxicity and the effectiveness of photodynamic cancer therapy. The two photodynamic complexes studied, [Ru(tpy)(pydppn)](2+), where pydppn = (3-(pyrid-2'-yl)-4,5,9,16-tetraaza dibenzo[a,c]naphthacene, and [Ru(pydppn)(2)](2+), differed in their efficiency of p53 and PCNA photocrosslinking in cells, but showed similar efficiency of photocrosslinking in cell lysates, suggesting that they differ in their ability to enter cells. Photocrosslinking of PCNA by [Ru(tpy)(pydppn)](2+) increased linearly with concentration, time of uptake, or light exposure. Both [Ru(tpy)(pydppn)](2+) and [Ru(pydppn)(2)](2+) caused photodynamic protein-DNA crosslinking in cells, but [Ru(tpy)(pydppn)](2+) was more efficient. The efficiency of photodynamic protein-DNA crosslinking by [Ru(tpy)(pydppn)](2+) in cells increased with increasing levels of photodynamic damage. Photodynamic damage by [Ru(tpy)(pydppn)](2+) caused inhibition of DNA replication in a classical biphasic response, suggesting that DNA damage signaling and cell cycle checkpoint pathways were still operative after significant damage to nuclear proteins. PMID- 20023927 TI - Structure-related frustrated magnetism of nanosized polyoxometalates: aesthetics and properties in harmony. AB - The structural versatility characterizing polyoxometalate chemistry, in combination with the option to deliberately use well-defined building blocks, serves as the foundation for the generation of a large family of magnetic clusters, frequently comprising highly symmetric spin arrays. If the spin centers are coupled by antiferromagnetic exchange, some of these systems exhibit spin frustration, which can result in novel magnetic properties of purely molecular origins. We discuss here the magnetic properties of selected nanosized polyoxometalate clusters featuring spin triangles as their magnetic 'building blocks' or fragments. This includes unique porous Keplerate clusters of the type {(Mo)Mo(5)}(12)M(30) (M = Fe(III), Cr(III), V(IV)) with the spin centers defining a regular icosidodecahedron and the {V(15)As(6)}-type cluster sphere containing a single equilateral spin triangle; these species are widely discussed and studied in the literature for their role in materials science as molecular representations of Kagome lattices and in relation to quantum computing, respectively. Exhibiting fascinating and unique structural features, these magnetic molecules allow the study of the implications of frustrated spin ordering. Furthermore, this perspective covers the impact of spin frustration on the degeneracy of the ground state and related problems, namely strong magnetic anisotropy and the interplay of antisymmetric exchange and structural Jahn-Teller effects. PMID- 20023928 TI - Multidentate fluorinated alkoxide ligand platforms for oxophilic metal centers: from MOCVD source reagents to polymerization catalysts. AB - In contrast to simple fluorinated alkoxides, the coordination chemistry of multidentate ligands that incorporate OC(CF(3))(2)CR(2)-type moieties and additional donors (N, O, etc.) has been briefly investigated. In this Perspective, we review some well-defined, unambiguously authenticated main group and transition metal complexes supported by multidentate fluorinated alkoxide ligands and we present aspects of their syntheses, structures, and reactivities. The first part is devoted to the first syntheses of fluorinated alkoxy-imino and fluorinated alkoxy-amino ligand platforms and their application in the preparation of late transition (Ru, Co, Ir, Ni, Pd, Cu) and main group (Sr, Ba, Ga) metal complexes, many of which have been used as chemical vapor deposition (CVD) source reagents. In the second part, heteroleptic complexes based on oxophilic metals (Y, La, Ti, Zr, Hf, Al) and the catalytic performance of these systems in olefin polymerization and ring-opening polymerization (ROP) of cyclic esters are highlighted. PMID- 20023929 TI - Unexpected pincer-type coordination (kappa3-SBS) within a zerovalent platinum metallaboratrane complex. AB - The first structurally characterised zerovalent platinum complex to contain a tridentate pincer-type coordination mode (kappa(3)-SBS) is presented, raising further questions concerning the geometries and trans influence of Z-type ligands. PMID- 20023930 TI - Unexpected formation of a novel pyridinium-containing catecholate ligand and its manganese(III) complex. AB - Nucleophilic aromatic substitution of tetrachloro-o-benzoquinone by pyridine and reduction of the o-quinone to the catechol by hydroxylamine forms 1,2-dihydroxy 3,5,6-trichlorobenzene-4-pyridinium chloride. This compound reacts with manganese(II) acetate in air to form chlorobis(3,5,6-trichlorobenzene 4 pyridinium catecholate)manganese(III), which represents the first complex of this ligand class to be structurally characterized by X-ray diffraction; this complex is active in the catalytic reduction of dioxygen to hydrogen peroxide under ambient conditions and turnover frequencies (TOFs) >10,000 h(-1) can be obtained. PMID- 20023931 TI - 3D Mn(II) coordination polymer with alternating azide/azide/formate/formate bridged chains: synthesis, structure and magnetic properties. AB - A new 3D Mn(II) coordination polymer, [Mn(2)(HCO(2))(L)(2)(N(3)) x MeOH](infinity) (1) (L = nicotinate N-oxide), was synthesized by a solvothermal reaction and characterized magnetically. Complex 1 is a typical antiferromagnet with alternating azide/azide/formate/formate bridges, exhibiting a new J(1)J(1)J(2)J(2)- coupling sequence. This is the first complex to contain an azide, formate and another type of carboxylate ligand. PMID- 20023932 TI - New alkylzinc complexes with bulky tris(triazolyl)borate ligands: surprising water stability and reactivity. AB - Bulky alkylzinc complexes, (Ttz(tBu,Me))ZnMe and (Ttz(tBu,Me))ZnEt, show remarkable stability towards water and both complexes crystallize with one molecule of adventitious water hydrogen bonded to two triazole rings; in contrast the less bulky complex, (Ttz(Ph,Me))ZnEt, reacts with water to yield (Ttz(Ph,Me))(2)Zn. PMID- 20023933 TI - Potassium-mediated zincation of ferrocene and ruthenocene: potassium, the architect behind supramolecular structural variations. AB - Direct zincation of ferrocene and ruthenocene by the synergic base [PMDETA x K(mu TMP)(mu-Me)Zn(Me)] produces the monozincated complexes [{PMDETA x K(mu Me)(2)Zn(Fc)}(infinity)] and [{PMDETA x K(mu-Me)(2)Zn(Rc)}(2)], respectively, having similar monomeric (dinuclear) units but aggregating supramolecularly in very different polymeric and dimeric forms. PMID- 20023934 TI - Stepwise assembly of a bis-hydroxylaminato gallium-indium complex. AB - A heterobimetallic gallium-indium complex with five-coordinate InMe group bonded via two O and two N atoms to a [GaR(2)ON(Me)(CH(2))(2)N(Me)O](2) macrocycle resulted from the reaction of the OH- and NH-functional bishydroxylaminato complex of gallium, MeN(OH)(CH(2))(2)N(H)(Me)OGaR(2)O cyclo[N(Me)(CH(2))(2)N(Me)OGaR(2)] (R = (t)Bu) with InMe(3), whereas the reaction of the latter with Me(3)N x AlH(3) affords a gallium hydride with Ga-Ga bonds. PMID- 20023935 TI - A new binding geometry for an ortho-xylylene-linked bis(NHC)cyclophane: a ruthenium(II) complex with a chelating (eta(1)-NHC)2:eta(6)-arene ligand. AB - Using two different reaction procedures, a Ru(II) complex has been isolated that contains an ortho-xylylene-linked bis(NHC)cyclophane (NHC = N-heterocyclic carbene) that binds to the Ru centre through two carbene carbons and one of the arene rings in an eta(6)-mode. PMID- 20023936 TI - Pt(II) complexes of unsubstituted guanine and 7-methylguanine. AB - In solution, unsubstituted guanine (GH2) is in a tautomeric equilibrium between 1H,7H-keto and 1H,9H-keto forms. Upon coordination with (dien)Pt(II), N9-bound mononuclear complex is preferentially formed. With excess (dien)Pt(II), the dinuclear complex [{Pt(dien)}2(GH-N7,N9)]3+ is formed at basic pH. The X-ray crystal structure analysis of mononuclear complexes [Pt(dien)(GH2-N9)](ClO4)2 x 2.25 H2O (1a), [Pt(dien)(GH2-N9)](Cl)(ClO4) (1b), and the dinuclear complex [{Pt(dien)}2(GH-N7,N9)](ClO4)3 (3) are reported. The N7 linkage isomer [Pt(dien)(GH2-N7)]2+ (2) could not be isolated and was tentatively assigned in solution. In HCl-acidic medium, the dinuclear complex 3 dissociates to form N9 bound mononuclear complex (1) as the major product and presumably the N7 linkage isomer (2) as a minor side product. Species 1 and 3 have been identified by the pD dependence of the 1H NMR guanine resonances. pKa values (calculated for H2O) for deprotonation of the N9 linkage isomer (1a, 1b) are 6.51 +/- 0.01 and 10.53 +/- 0.05 as well as 0.52 +/- 0.07 and 8.55 +/- 0.06 for the dinuclear complex [{Pt(dien)}2(GH-N7,N9)]3+ (3). Ab initio calculations have been performed for [Pt(dien)(GH2-N7)]2+, [Pt(dien)(GH2-N9)]2+ and [{Pt(dien)}2(GH-N7,N9)]3+. They reveal that in gas phase the N7 linkage isomer is more stable than the N9 linkage isomer by DeltaE = 8.4 kcal mol(-1). Reactions with (dien)Pt(II) and cis (NH3)2Pt(II) were also carried out to study the metal binding behaviour of the ligand, 7-methylguanine (7-MeGH). The complexes [Pt(dien)(7-MeGH-N9)](NO3)(ClO4) (4), cis-[Pt(NH3)2(7-MeGH-N9)2]Cl2 x 2 H2O (5) and cis,cis-[{PtCl(NH3)2}2(7-MeGH N3,N9)]Cl2 x 3 H2O (6) have been characterized by X-ray crystallography. pKa values for 4 are < 2 and 8.16 +/- 0.04, and for 5 1.86 +/- 0.45 and 8.00 +/- 0.08 (average for deprotonation of two 7-MeGH ligands). Structural peculiarities of N9 platinated guanine ligands are discussed and pKa values of Pt complexes with related guanine nucleobases are compared. PMID- 20023937 TI - Syntheses and characterization of bis(trifluoromethyl)phosphino naphthalenes and acenaphthenes. AB - Syntheses of heteroleptic 1,8-bis(phosphino)naphthalenes and 5,6 bis(phosphino)acenaphthenes were attempted using several synthetic strategies. Reaction of aryllithium with triphenylphosphite gave ArP(OPh)(2) (Ar = substituted naphthalene or acenaphthene), which was transformed into ArP(CF(3))(2), using a nucleophilic trifluoromethylation reaction with Me(3)SiCF(3)/CsF. The importance of the correct choice of solvent for the trifluoromethylation reactions is discussed. The incompatibility of ArP(CF(3))(2) with organolithium hampered the attachment of the second phosphine functionality to the organic backbone. Tetraphenoxyethylene was obtained in a small amount as a side product in the trifluoromethylation reaction. Selected new compounds were characterized by single-crystal X-ray diffraction. PMID- 20023938 TI - Thermal stability of alkali and alkaline-earth substituted LAMOX oxide-ion conductors. AB - The high temperature demixing/recombination phenomenon previously observed in Ca substituted La(2)Mo(2)O(9) oxide ion conductors [A. Selmi et al., Solid State Ionics, 2006, 177, 3051; Eur. J. Inorg. Chem., 2008, 1813] has been visualised using scanning electron microscopy and EDX analysis. The demixed state appears as CaMoO(4) straight solid streams erupted from pores within LAMOX grains. The thermal stability study is extended to other alkali and alkaline-earth substituted LAMOX compounds, all of which are shown, in temperature-controlled X ray diffractograms, to present similar demixing/recombination processes. The most spectacular effect is observed in La(1.88)K(0.12)Mo(0.6)W(1.4)O(8.88) where demixing takes the form of a total decomposition, before full recombination at a higher temperature. Such a phenomenon is interpreted as originating from temperature-dependent solid solution limits with higher substitutional ranges at higher temperatures. It results in the metastabilisation of pure phases by quenching (or rapid cooling), whereas the stable state is demixed, as shown on slowly cooled samples. PMID- 20023939 TI - Activation of C-H bonds mediated by Mo[triple bond]Mo moieties in heterobimetallic Zn/O/Mo clusters. AB - Heterobimetallic drive and cooperativity lead to a striking scenario for C-H bond activation and heteroaggregation involving Mo(III) dimers. Reaction of (RO)(3)Mo[triple bond]Mo(OR)(3) with molar excess of ZnMe(2) at-78 degrees C affords the novel mixed Zn, Mo oxo clusters [Mo(2)(MeZn)(6)(mu(2)-Me)(mu(3) CH(2))(2)(OR)(7)] 1 (R = neopentyl) and 2 (R = cyclohexyl) preserving the Mo[triple bond]Mo triple bond, while the same reaction performed at ambient temperature gives rise to the related cluster [Mo(2)(MeZn)(6)(mu(2)-CH(2))(mu(3) CH(2))(2)(OR)(6)] 3. These complexes represent the first examples of heterobimetallic Mo[triple bond]Mo complexes with bridging methyl and methylene groups and agostic C-H-->Mo interactions, as well as penta-coordinate carbon. The complexes 1-3 were characterized by elemental analyses, multinuclear NMR and single-crystal X-ray diffraction analysis. PMID- 20023940 TI - Hydrothermal syntheses, crystal structures and magnetic properties of two new mixed metal copper(I) sulfites. AB - Two new examples of mixed metal copper(I) sulfites, namely, Na3Mn2Cu(SO3)4(H2O)5 (1) and NaMn4Cu(SO3)5(H2O)3 (2), have been synthesized and structurally characterized. The 1D structure of 1 is built from a chain of [Mn2Cu(SO3)4]3- bridged by Na+ ions whereas the structure of 2 features a complicated 3D framework in which Mn8O36 octanuclear clusters are bridged by sulfite anions and copper(I) ions into a 3D [CuMn4(SO3)5(H2O)3]2- network. The eight-coordinated sodium(I) ions are located at the cavities of the 3D structure. In both compounds, the oxygen atoms of the sulfite anion are bonded to Mn(II) ions whereas the sulfur atom is coordinated to the Cu+ ion. Magnetic measurements indicate that there exists antiferromagnetic interactions between Mn(II) centers in both compounds. PMID- 20023941 TI - High pressure studies of hydroxo-bridged Cu(II) dimers. AB - A combination of high pressure single crystal X-ray diffraction and high pressure SQUID magnetometry has been used to study three hydroxo-bridged copper(II) dimers. [Cu2(OH)2(H2O)2(tmen)2](ClO4)2 (1; tmen = tetramethylethylenediamine), [Cu2(OH)2(tben)2](ClO4)2 (2; tben = di-tbutylethylenediamine) and [Cu2(OH)2(bpy)2](BF4)2 (3; bpy = 2,2'-bipyridine) have been structurally determined to 2.5, 0.9 and 4.7 GPa, respectively. The application of hydrostatic pressure imposes significant distortions and modifications in the structures of all three complexes. This is particularly true of the bond distances and angles between the metal centres and the bridging hydroxo groups. Compound 1 undergoes a phase transition between 1.2 and 2.5 GPa caused by the loss of a coordinated water molecule. This leads to a loss of symmetry and dramatic changes in the molecular structure of the complex. The structural changes are manifested in changes in the magnetic behaviour of the complexes as seen in dc susceptibility measurements up to approximately 0.9 GPa for 1, 2 and 3: the exchange becomes less antiferromagnetic in 1 and 2 and more ferromagnetic in 3. PMID- 20023942 TI - Synthesis and structural and magnetic characterisation of copper(II) complexes of mixed phosphonate-antimonate ligands. AB - A polynucleating oxygen donor ligand based on condensation of p chlorophenylstibonic acid (ArSbO(3)H(2)) and t-butylphosphonic acid is reported. [(SbAr)(2)O(HO(3)P(t)Bu)(6)] contains two antimony centres, bridged by an oxide and two hydrogen phosphonates. Reaction with copper acetate under solvothermal conditions produces four new polymetallic copper complexes. With pyridine used as a base the major product is a tetracopper cage, [Cu(4)O(2)(SbAr)(2)(O(3)P(t)Bu)(2)(O(2)CMe)(2)(OMe)(6)], with the four copper centres arranged in a rhombus; a minor product is a tricopper complex, [Cu(3)O(4)(SbAr)(2)(O(3)P(t)Bu)(4)(py)(3)]. With LiOMe as base the major product is again the tetranuclear cage, but the minor product is a heterometallic cage, [Cu(5)Li(4)O(6)(SbAr)(4)(O(3)P(t)Bu)(6)(O(2)CMe)(2)(OMe)(4) (MeOH)(4)]. With 2,6 lutidine as base an octametallic complex is formed, [Cu(8)O(4)(SbAr)(2)(O(3)P(t)Bu)(6)(O(2)CMe)(4)(lutidine)(2)]. Magnetic studies show both anti-ferromagnetic and weak ferromagnetic exchange between the copper centres in these new complexes. PMID- 20023943 TI - Reactivity of germanium(II) hydride with nitrous oxide, trimethylsilyl azide, ketones, and alkynes and the reaction of a methyl analogue with trimethylsilyl diazomethane. AB - The reactions of stable beta-diketiminate germanium(II) hydride LGeH (1) [L = HC(CMeNAr)2, Ar = 2,6-iPr2C6H3] with nitrous oxide, trimethylsilyl azide, ketones, and alkynes are described. 1 reacts with nitrous oxide to yield the germanium(II) hydroxide LGeOH (2), and with trimethylsilyl azide affords in toluene at room temperature the germanium(II) azide LGeN3 (3), and also the germanium(IV) diamide L'Ge(NHSiMe3)2 (L' = CH{(CCH2)(CMe)(2,6-iPr2C6H3N)2}) (4). Ketones (PhCOCF3, 2-C4H3SCOCF3) and 1 generated the germanium(II) alkoxides (5-6) in high yield. The activated terminal alkyne (HC[triple bond]CCO2Me) and disubstituted alkyne (EtO2CC[triple bond]CCO2Et) react with 1 to form the germanium(II) substituted alkenes (vinyl germylene) (7-8). Further reaction of the methylgermanium(II) compound LGeMe (9) with trimethylsilyl diazomethane resulted in the formation of germanium(IV) amide L'Ge(Me)NHN=CHSiMe3 (10). Compounds 2-8, and 10 were characterized by microanalysis and multi-nuclear NMR spectroscopy. Furthermore compounds 3-6, and 8 are confirmed by X-ray structural analysis. PMID- 20023944 TI - Synthesis and characterization of xanthene-bridged Schiff-base dimanganese(III) complexes: bimetallic catalysts for asymmetric oxidation of sulfides. AB - Dimanganese(III) complexes of salen-type ligands anchored by 9,9-dimethylxanthene 4,5-diyl spacers were synthesized. Two types of structures, cyclic and acyclic forms, are presented. The 2 + 2 Schiff-base condensation of 5,5'-(9,9 dimethylxanthene-4,5-diyl)bis(salicylaldehyde) and 1,2-diaminobenzene gave a macrocyclic ligand, from which a cyclic dimanganese(III) complex was synthesized. A similar dimanganese(III) complex with stereogenic centers was synthesized by the metal-assisted condensation reaction using (1R,2R)-1,2-diaminocyclohexane. Acyclic dimanganese(III) complexes anchored by one xanthene spacer were obtained by metal-assisted stepwise condensation reactions using the xanthene-bridged bis(salicylaldehyde), (1R,2R)-1,2-diaminocyclohexane monohydrochloride, and salicylaldehydes. X-Ray crystal structure analysis confirmed that both cyclic and acyclic complexes have two cofacially oriented salen-type manganese(III) units in a complex cation, showing an Mn...Mn distance of ca. 5.1 A. Asymmetric oxidation of sulfides was performed by using the chiral dimeric complexes as catalysts, which showed enantiomeric excesses ranging from 5 to 19%. The addition of 4 (dimethylamino)pyridine to the reaction system improved the enantioselectivity up to 39% ee. This effect was not observed for the corresponding mononuclear catalyst. PMID- 20023945 TI - Spin crossover in iron(III) Schiff-base 1-D chain complexes. AB - A series of iron(III) 1-D polymeric materials of the general formula [Fe(III)(Schiff-base)(L)](BPh(4)).n(CH(3)OH) (Schiff base = N,N' ethylenebis(salicylaldimine) (H(2)salen), N,N'-o-phenylenebis(salicylaldimine) (H(2)salophen) and N,N'-ethylenebis(acety1acetone)-2,2'-imine (H(2) acen); L = bridging di-pyridyl or di-imidazole ligand, n = 0-4) and analogues therein, have been synthesised and structurally and magnetically characterised. In this series, a range of structural motifs are observed including linear 1-D chains, hydrogen bonded chains, a 'hybrid' 1-D chain- and -dimer compound and a hydrogen-bonded dinuclear material; all exhibit extensive intermolecular interactions. The magnetic consequences of varying both the equatorial Schiff-base ligands and axial bridging ligands have been investigated. Overall, we find that, independent of the axial bridging ligand employed, the salen equatorial ligand results in a high spin character and the acen ligand results in spin crossover character, generally with a spin transition of a gradual nature. Variations in magnetic behaviour can be rationalized, in part, in terms of the C(2)N(2) backbone conformation of the equatorial Schiff base ligand, which may either inhibit or allow a spin transition. PMID- 20023946 TI - Y(BH4)3--an old-new ternary hydrogen store aka learning from a multitude of failures. AB - Fourteen different synthetic approaches towards pure solvent-free Y(BH(4))(3) have been tested, thirteen of which have failed. Attempted reactions of YCl(3) or Y(OC(4)H(9))(3) with LiBH(4) in THF, those of YCl(3) with (C(4)H(9))(4)N(+) BH(4)(-), as well as between YH(x approximately 3) and R(4)NBH(3) (R = CH(3), C(2)H(5)) in the presence or absence of a solvent (n-hexane or CH(2)Cl(2)) did not lead to the expected product. The mechanochemical solid/solid reactions (MBH(4) + 3 YX(3)--> Y(BH(4))(3) + 3 LiCl, where M = Li, Na; X = F, Cl) have succeeded only for the LiBH(4) and YCl(3) reagents, but the separation of the crystalline reaction products (Y(BH(4))(3) in its Pa3 phase and LiCl) by dissolution or flotation in various solvents has not been successful. The thermal decomposition process of Y(BH(4))(3) in a mixture with LiCl has been investigated with thermogravimetric (TGA) and calorimetric analysis (DSC) combined with spectroscopic evolved gas analysis (EGA). Three major endothermic steps could be distinguished in the DSC profile at ca. 232, 282, 475 degrees C (heating rate 10 K min(-1)) corresponding to a phase transition and two steps of thermal decomposition. Solid decomposition products are amorphous except for the new cubic polymorph of Y(BH(4))(3) overlooked in previous work. The high-temperature phase forms at the onset of thermal decomposition and it may be prepared by heating of the low-temperature phase up to a narrow temperature range (194-210 degrees C) followed by rapid quenching. Y(BH(4))(3) constitutes a novel highly efficient hydrogen storage material (theor. 9.0 wt% H) but, unfortunately, the evolved H(2) is contaminated by toxic boron hydrides and products of their pyrolysis. PMID- 20023947 TI - Dinuclear, tetranuclear and one-dimensional pyrazine-based copper(II) complexes: preparation, X-ray structure and magnetic properties. AB - The preparation, crystal structures and magnetic properties of the copper(II) complexes of formula [Cu(2)(tppz)(H(2)O)(2)(CF(3)SO(3))(4)] (1), [Cu(tppz)(CrO(4))](n) x 3nH(2)O (2) and [Cu(4)(tppz)(4)(H(2)O)(4)(MoO(4))(2)](CF(3)SO(3))(4) x 7 H(2)O (3) [tppz = 2,3,5,6-tetrakis(2-pyridyl)pyrazine] are reported. Complex 1 is a dinuclear species where tppz adopts a bis-tridentate bridging mode with an intramolecular copper-copper separation of 6.5221(6) A. Each copper(II) ion in 1 has an elongated octahedral geometry with three tppz nitrogen atoms and a water molecule in the equatorial positions and two triflate oxygen atoms occupying the axial sites. Complex 2 is a uniform copper(II) chain where the copper atoms are bridged by two oxygen atoms of the chromate group (mu(1,2)-CrO(4)), the value of the intrachain metal-metal separation being 4.2614(5) A. Each copper(II) ion in 2 exhibits a somewhat distorted square pyramidal geometry with three nitrogen atoms from a tridentate tppz ligand and a chromate oxygen atom in the basal plane and an oxygen atom of a symmetry-related chromate group in the apical position. Complex 3 is a cyclic tetranuclear compound where peripheral [Cu(tppz)(H(2)O)](2+) units are connected through two molybdate groups, each one adopting the mu(1,1,2)-MoO(4) bridging mode. Regular alternating five [Cu(1) and Cu(4)] and six [Cu(2) and Cu(3)] coordinated copper(II) ions occur in 3 with distorted square pyramidal CuN(3)O(2) and elongated octahedral CuN(3)O(3) surroundings, respectively. The four copper atoms in 3 have in common the presence of a molybdate-oxygen and three tppz-nitrogen atoms in the basal [Cu(1) and Cu(4)]/equatorial [Cu(2) and Cu(3)] planes and a water molecule in the apical [Cu(1) and Cu(4)]/one axial [Cu(2) and Cu(3)] positions whereas the remaining axial position is occupied by a molybdate-oxygen [Cu(2) and Cu(3)]. The shorter intramolecular copper-copper separation is 3.371(1) A [Cu(2)...Cu(3)], the other ones varying in the range 5.537(1)-6.301(1) A. The investigation of the magnetic properties of 1-3 in the temperature range 1.9-295 K has shown the occurrence of intermediate antiferro- [J = -48.0(1) cm(-1) with H = -JS(1) x S(2) (1)], weak ferro- [J = +0.28(1) cm(-1) with H = -JSigma(i)S(i) x S(i+I) (2)] and coexistence of weak ferro- and antiferromagnetic interactions [J(1) = +2.38(2) cm(-1), J(2) = +0.56(2) cm(-1) and J(3) = -1.53(2) cm(-1) with H = -J(1) (S(2) x S(3)) + J(2) (S(1) x S(3) + S(2) x S(4)) + J(3) (S(1) x S(2) + S(3) x S(4)) (3)] between the copper(II) ions across bis-tridentate tppz (1), mu(1,2)-CrO(4) (2), di-mu(1,1) MoO(4) and mu(1,2)-MoO(4) (3) bridges. The values of the magnetic interactions are analyzed in terms of simple orbital symmetry considerations and compared with those reported for parent systems. PMID- 20023948 TI - Dipyrrin based luminescent cyclometallated palladium and platinum complexes. AB - A series of complexes based on the combination of cyclometallated palladium or platinum moieties with functionalized dipyrrin ligands bearing mesityl- or benzonitrile groups have been prepared and characterized both in the solid state and in solution; these compounds exhibit a characteristic dipyrrin-centered luminescence with an emission intensity modulated by the degree of rotational freedom of the aromatic group attached to the dipyrrin chelate. PMID- 20023949 TI - Densely packed Gd(III)-chelates with fast water exchange on a calix[4]arene scaffold: a potential MRI contrast agent. AB - A pyridine-N-oxide functionalized DOTA analogue has been conjugated to a calix[4]arene and the corresponding Gd-complex was characterized with respect to its suitability as MRI contrast agent. The compound forms spherical micelles in water with a cmc of 35 microM and a radius of 8.2 nm. The relaxivity of these aggregates is 31.2 s(-1) mM(-1) at 25 degrees C and 20 MHz, which corresponds to a molecular relaxivity of 125 s(-1) mM(-1). The high relaxivity mainly originates from the short tau(M) (72.7 ns) and the size of the micelles. The interaction with bovine serum albumin (BSA) was studied and an observed relaxivity of up to 40.8 s(-1) mM(-1) (163.2 s(-1) mM(-1) per binding place) at 20 MHz and 37 degrees C was found in the presence of 2.0 mM protein. PMID- 20023950 TI - A Mn(III) triplesalen-based 1D pearl necklace: exchange interactions and zero field splittings in a C3-symmetric Mn(III)6 complex. AB - The reaction of the tert-butyl-substituted triplesalen ligand H(6)talen(t-Bu(2)) with 2.8 equivalents of Mn(OAc)(2) x 4 H(2)O in MeOH in the presence of NaBPh(4) results in the formation of the one-dimensional (1D) coordination polymer {[{(talen(t-Bu(2)))Mn(3)(MeOH)}(2)(mu(2)-OAc)(3)](mu(2)-OAc)}(n)(BPh(4))(2n) ({[Mn(III)(6)](OAc)}(n)(BPh(4))(2n)) which has been characterized by FTIR, elemental analysis, ESI-MS, single-crystal X-ray diffraction and magnetic measurements. The triplesalen ligand (talen(t-Bu(2)))(6-) provides three salen like coordination compartments bridged in a meta-phenylene arrangement by a phloroglucinol backbone resulting in the trinuclear Mn(III) base unit {(talen(t Bu(2)))Mn(3)}(3+). Two of these base units are bridged by three inner acetate ligands giving rise to the hexanuclear complex [{(talen(t Bu(2)))Mn(3)(MeOH)}(2)(mu(2)-OAc)(3)](3+) ([Mn(III)(6)](3+)). These complexes are bridged by a single external acetate to form a 1D chain as pearls in a pearl necklace. Variable temperature-variable field and mu(eff)vs. T magnetic data have been analyzed in detail by full-matrix diagonalization of the appropriate spin Hamiltonian consisting of isotropic exchange, zero-field splitting, and Zeeman interaction taking into account the relative orientation of the D-tensors. Satisfactory reproduction of the experimental data have been obtained for parameters sets J(1) = -(0.60 +/- 0.15) cm(-1), J(2) = -(1.05 +/- 0.15) cm(-1), and D(Mn) = -(3.0 +/- 0.7) cm(-1) with J(1) describing the exchange through the phloroglucinol backbone and J(2) describing the exchange through the inner acetates. The non-necessity to incorporate the bridging outer acetates correlates with the longer Mn-O bonds. The experimental data can neither be analyzed without incorporating zero-field splitting nor by the application of a single effective spin ground state. PMID- 20023951 TI - Trinuclear ruthenium dioxolene complexes based on the bridging ligand hexahydroxytriphenylene: electrochemistry, spectroscopy, and near-infrared electrochromic behaviour associated with a reversible seven-membered redox chain. AB - The trinuclear complexes [{(R2bipy)2Ru}3(mu3-HHTP)](PF6)3 [1(PF6)3, R = H; 2(PF6)3, R = 4-tBu] contain three {Ru(R2bipy)2}2+ fragments connected to the triangular tris-chelating ligand hexahydroxytriphenylene (H6HHTP). This bridging ligand contains three dioxolene-type binding sites, each of which can reversibly convert between dianionic catecholate (cat), monoanionic semiquinone (sq) or neutral quinone (q) redox states. The bridging ligand as a whole can therefore exist in seven different redox states from fully reduced [cat,cat,cat]6- through to fully oxidised, neutral [q,q,q]. Cyclic voltammetry of 1(PF6)3 in MeCN reveals six redox processes of which the three at more positive potentials (the sq/q couples) are reversible but the three at more negative potentials (the sq/cat couples) are irreversible with distorted wave shapes due to the insolubility of the reduced forms of the complex. In contrast, the more soluble complex 2(PF6)3 displays six reversible one-electron redox processes making all components of a seven-membered redox chain accessible. UV/Vis/NIR spectro-electrochemical studies reveal rich spectroscopic behaviour, with--in particular--very intense transitions in the near-IR region in many of the oxidation states associated with Ru(II)-->(dioxolene) MLCT and bridging ligand centred pi-pi* transitions. TDDFT calculations were used to analyse the electronic spectra in all seven oxidation states; the calculated spectra generally show very good agreement with experiment, which has allowed a fairly complete assignment of the low-energy transitions. The strong electrochromism of the complexes in the near-IR region has formed the basis of an optical window in which a thin film of 1(PF6)3 or 2(PF6)3 on a conductive glass surface can be reversibly and rapidly switched between redox states that alternate between strongly absorbing or near transparent at 1100 nm, with--for 2(PF6)3--the switching being stable and reversible in water over thousands of cycles. PMID- 20023952 TI - Vanadate complexes in serum: a speciation modeling study. AB - The speciations of two drug candidate ligands, 2-hydroxypyridine-N-oxide (Hhpno) and 2-mercaptopyridine-N-oxide (Hmpno), with vanadate (V(V)) were determined at 25.0 degrees C and 0.20 mol dm(-3) KCl by pH-metric and (51)V-NMR methods. At pH 7.4, the two predominant compounds with both ligands are the VO(2)L(2) and VO(2)L(OH). NH(4)[VO(2)(hpno)(2)] x 3 H(2)O was prepared in solid form, and its crystal structure was determined by X-ray diffraction. The stabilities of the complexes VO(2)L(2) of five drug candidate ligands were compared at pH 7.4. In view of the stability sequence hpno > maltol approximately hdp (Hhdp: 3-hydroxy 1,2-dimethyl-4-pyridinone) >> mpno > picolinic acid, the first two of these ligands were chosen for equilibrium studies with apotransferrin (apoTf) competition. The V(V)-apoTf stability constants (log K(1) = 6.03 +/- 0.10; log K(2) = 5.46 +/- 0.18) determined by (51)V-NMR spectroscopy were confirmed by ultrafiltration. Both methods proved that there seems to be no hydrogencarbonate vanadate competition for the apoTf anion-binding positions. The other potential high molecular mass V(V) binder in the serum is human serum albumin (HSA). As no interaction was detected by (51)V-NMR spectroscopy or fluorimetry, the binding properties of HSA were quantified on the basis of literature data. As a final conclusion, speciation modeling calculations suggest that, under serum conditions, apoTf is probably the primary metal ion binder, even in the presence of the most stable V(V) carrier ligands hpno and maltol and HSA plays a negligible role in V(V) binding. PMID- 20023953 TI - Isocyanate diinsertion into the N-H bond of the 2-pyridylamino ligand of organolanthanides. AB - [Cp2LnNHPy]2 (Py = 2-pyridyl) (1a-e) react with phenyl isocyanate to form the N-H diinsertion products Cp2Ln[eta2:eta1-PyNCON(Ph)CONHPh](THF) (Ln = Yb (3a), Er (3b), Y (3c), Dy (3d), Gd (3e)). It has been proven that nPr2NH can abstract one PhNCO unit from 3c to form Cp2Y[eta3-OC(NHPh)NPy] (2c) and nPr2NHCONHPh (4), representing a rare example of selective release of a functional group of ligands in organolanthanide chemistry. Hydrolysis of 2c gives the organic nitrogen containing product PyNHCONHPh (5). Moreover, 3c can also be obtained by the reaction of 2c with PhNCO. These results demonstrate that the diinsertion of PhN=C=O into the N-H bond of coordinated amino ligands might proceed in a stepwise manner. All the compounds were characterized by elemental analysis and spectroscopic properties. The structures of compounds 3a-e and 4 are also determined through X-ray single-crystal diffraction analysis. PMID- 20023954 TI - Synthesis of InP nanofibers from tri(m-tolyl)phosphine: an alternative route to metal phosphide nanostructures. AB - The synthesis of InP nanofibers via a new Ullmann-type reaction of indium nanoparticles with tri(m-tolyl)phosphine (P(PhMe)(3)) was typically performed to illustrate an alternative route for the preparation of nanostructured metal phosphides, including III-V (13-15) and transition-metal phosphides. Triarlyphosphine compounds such as other two tri(m-tolyl)phosphine isomers, diphenyl(p-tolyl)phosphine, and triphenylphosphine were comparably employed to synthesize InP nanocrystals. From the aspect of the carbonization of triarlyphosphines, Raman spectroscopy and thermo-gravimetric analysis (TGA) investigations of the InP products showed that the stability of these triarlyphosphines conformed to the order of tri(p-tolyl)phosphine approximately tri(o-tolyl)phosphine < diphenyl(p-tolyl)phosphine < tri(m-tolyl)phosphine < triphenylphosphine. The correlation between the stability of triarlyphosphines and the growth of InP nanocrystals was investigated, and experimental results showed that the relatively stable triarlyphosphines (tri(m-tolyl)phosphine and triphenylphosphine) were favorable for the preparation of one-dimensional (1D) InP nanostructures (nanofibers and nanowires). The reactivity (stability) of triarlyphosphines was also compared with those of P(SiMe(3))(3) (typically see: J. M. Nedeljkovic, O. I. Micic, S. P. Ahrenkiel, A. Miedaner and A. J. Nozik, J. Am. Chem. Soc., 2004, 126, 2632) and P(C(8)H(17))(3) (C. Qian, F. Kim, L. Ma, F. Tsui, P. D. Yang and J. Liu, J. Am. Chem. Soc., 2004, 126, 1195) according to the difference in preparative temperature for phosphide synthesis. Raman and photoluminescence properties of the as-synthesized InP nanocrystals were further studied, and the synthetic mechanism of our method was reasonably investigated by GC-MS analysis. Moreover, the current route was successfully extended to prepare GaP, MnP, CoP and Pd(5)P(2) nanocrystals. PMID- 20023955 TI - Synthesis of phosphine substituted beta-diketiminate based isomeric Ge(II) complexes. AB - Treatment of phosphine substituted beta-diketiminate lithium, Ph2PC[C(Me)N(Dipp)]2LiOEt2 (1) (Dipp = 2,6-iPr2C6H3) with dioxane x GeCl2 resulted in a five-membered N,P chelate complex, Ph2PC[C(Me)N(Dipp)]2GeCl (2). An isomer of 2 CH{[C(CH2PPh2)N(Dipp)][C(Me)N(Dipp)]}GeCl (5) was obtained from the germanium complex L'Ge (L' = CH[C(CH2)N(Dipp)][C(Me)N(Dipp)])Ge (4) and Ph2PCl. In 5 the PPh2 group remains uncoordinated. Both complexes were characterized by X ray structural analysis and in the case of 5 both enantiomers crystallize in the same unit cell. PMID- 20023956 TI - Platinum(II) and palladium(II) metallomacrocycles derived from cationic 4,4' bipyridinium, 3-aminopyrazinium and 2-aminopyrimidinium ligands. AB - A series of cationic, ditopic N-donor ligands based on 4,4'-bipyridine (4,4' bipy), 3-aminopyrazine (apyz) and 2-aminopyrimidine (apym), each incorporating two positively-charged N-heterocycles linked by a conformationally-flexible spacer unit, have been synthesised and treated with palladium(II) or platinum(II) precursors [M(2,2'-bipy)(NO(3))(2)] (M = Pd(II) or Pt(II)) to form highly cationic metallocyclic species. Treatment of 1,6-bis(4,4'-bipyridinium)hexane nitrate with [M(2,2'-bipy)(NO(3))(2)] in aqueous solution, followed by the addition of KPF(6), resulted in the formation of the [2+2] species [M(2)(2,2' bipy)(2){4,4'-bipy(CH(2))(6)4,4'-bipy}(2)](PF(6))(8). Treatment of [Pd(PhCN)(2)Cl(2)] with 1,3-bis(4,4'-bipyridinium)propane hexafluorophosphate in MeCN afforded [Pd(2)Cl(4){4,4'-bipy(CH(2))(3)4,4'-bipy}(2)](PF(6))(4). When the cationic apyz or apym ligands were used in aqueous solution, the analogous metallomacrocycles did not form. Instead, deprotonation of the exocyclic amino group occurred upon coordination of the ligand to afford a tetranuclear [4+2] species in the case of platinum(II), with Pt(II)...Pt(II) bonding supported by strong UV-vis absorption at lambda = 428 nm which was assigned to a metal-metal to-ligand charge transfer (MMLCT) band. Thus, treatment of 1,6-bis(3 aminopyrazinium)hexane nitrate with [Pt(2,2'-bipy)(NO(3))(2)], followed by the addition of KPF(6), led to the formation of the red species [Pt(4)(2,2' bipy)(4){apyz(CH(2))(6)apyz-2H}(2)](PF(6))(8). No related products could be identified with palladium(II), consistent with the low propensity for this metal ion to form strong Pd(II)...Pd(II) bonding interactions. PMID- 20023957 TI - Trilacunary A-beta-Keggin tungstogermanates and -silicates functionalized with phenyltin(IV) electrophiles. AB - The first organic derivative of a beta-Keggin tungstogermanate, the monomeric Keggin anion [{(C(6)H(5))Sn(OH)}(3)(A-beta-GeW(9)O(34))](4-) (1), was the only species isolated from the reaction of (C(6)H(5))SnCl(3) with Na(10)[A-beta GeW(9)O(34)] in water, whereas the Si-analogue [{(C(6)H(5))Sn(OH)}(3)(A-beta SiW(9)O(34))](4-) (2) and two dimeric tungstosilicates, the unprecedented species [{(C(6)H(5))Sn(A-beta-H(3)SiW(9)Sn(2)O(37))}(2)O(2)](8-) (3) and the known sandwich-type polyanion [{(C(6)H(5))Sn(OH)}(3)(A-beta-H(3)SiW(9)O(34))(2)](8-) (4), were isolated starting from [A-beta-SiW(9)O(34)](10-). The novel polyanion 3 can be described as the product of a double Sn-C hydrolysis followed by condensation of two subunits of 2 through double Sn-O-Sn bridge formation. Solid state characterization has been carried out by infrared spectroscopy, thermogravimetry and single-crystal X-ray diffraction, representing the first structural analysis for 4. A full solution multinuclear NMR ((1)H, (13)C, (117)Sn, and (183)W) and solid-state (117)Sn CP-MAS NMR study enabled full characterization of 1. A weak 1-dimensional polyanion association through (Sn)O HO(t) hydrogen contacts involving one of the protonated bridging O atoms of the (C(6)H(5))(3)Sn(3)O(12) corner-shared triad is observed for 1 by X-ray diffraction, resulting in a local C(s) pseudosymmetry for the Sn centers detected by solid-state (117)Sn NMR spectroscopy. PMID- 20023958 TI - Synthesis of 1,3-azaphosphol-2-ones. Crystal and molecular structures of [SP-4-2] dichlorobis(3-phenyl-1,3-dihydrobenzo[1,3]azaphosphol-2-one-P)palladium(II) and its chloro(methyl)platinum(II) analogue. AB - Reaction of secondary phosphine (+/-)-(2-aminophenyl)phenylphosphine, (+/-)-app, with PCl(5) in toluene gives the hydrochloride salt of the expected chlorophosphine (+/-)-(2-aminophenyl)chlorophenylphosphine, (+/-)-acpp.HCl, however, this is not the case with triphosgene. Rather the first example of a 1,3 azaphosphol-2-one is isolated, viz. (+/-)-3-phenyl-1,3 dihydrobenzo[1,3]azaphosphol-2-one, (+/-)-pbap. The hydrochloride salt (+/-) acpp.HCl readily reacts with excess vinyl-, 2-methylphenyl- or 2-methoxyphenyl magnesium bromide to give the corresponding tertiary phosphines (+/-)-(2 H(2)NC(6)H(4))PPhR (where R = CH=CH(2), 2-C(6)H(4)Me or 2-C(6)H(4)OMe). Hydrophosphination of the vinyl substituted tertiary phosphine with (+/-)-app in the presence of KOBu(t) provides a synthetic route to the elusive P(2)N(2) quadridentate ligand (R(P)*,R(P)*)- and (R(P)*,S(P)*)-(CH(2))(2)(PPhC(6)H(4)NH(2) 2)(2), albeit in low yield. The azaphospholone (+/-)-pbap can be readily deprotonated with KOBu(t) in thf and subsequently alkylated with methyl iodide or benzyl bromide to give the analogous N-methyl or N-benzyl derivatives. Alkylation with 1,3-dibromopropane gives the bis(azaphospholone) (R(P)*,R(P)*)- and (R(P)*,S(P)*)-1,3-bis[1-{3-phenyl-1,3-dihydrobenzo[1,3]azaphosphol-2 one}]propane. The latter and the N-methyl substituted azaphospholone can also be synthesised by the reaction of the corresponding secondary phosphine, viz. (R(P)*,R(P)*)- and (R(P)*,S(P)*)-(CH(2))(3)(NHC(6)H(4)PHPh-2)(2) and (+/-)-(2 methylaminophenyl)phenylphosphine, with triphosgene. All three azaphospholones react with [PtClMe(1,5-cyclooctadiene)] in thf to give complexes of the type cis [PtClMeL(2)] in which ligand L is coordinated via the P atom of the azaphospholones. The ligand (+/-)-pbap has also been complexed to palladium(II) via the reaction with Li(2)[PdCl(4)] in methanol to give cis-[PdCl(2){(+/-) pbap}(2)]. The structures of cis-[PtClMe{(+/-)-pbap}(2)] and cis-[PdCl(2){(+/-) pbap}(2)] have been confirmed by X-ray analysis. PMID- 20023959 TI - Dialkylamino cyclopentadienyl ruthenium(ii) complex-catalyzed alpha-alkylation of arylacetonitriles with primary alcohols. AB - Aminocyclopentadienyl ruthenium complexes, [(eta(5) C(5)H(4)NMe(2))Ru(PPh(3))(2)(CH(3)CN)](+)BF(4)(-) and [(eta(5) C(5)H(4)NEt(2))Ru(PPh(3))(2)(CH(3)CN)](+)BF(4)(-), are moderately active catalysts for alpha-alkylation of arylacetonitriles with primary alcohols; on the other hand, the analogous unsubstituted cyclopentadienyl ruthenium complex [(eta(5)-C(5)H(5))Ru(PPh(3))(2)(CH(3)CN)](+)BF(4)(-) shows very low catalytic activity. On the basis of experimental results and theoretical calculations, rationalization for the much higher catalytic activity of the aminocyclopentadienyl complexes over that of the unsubstituted Cp complex is provided. In the catalytic systems with the former, it is possible to regenerate the active solvento complexes via protonation of the metal hydride intermediates and subsequent ligand substitution; this process is, however, very nonfacile in the catalytic system with the latter. PMID- 20023961 TI - The contribution of computational studies to organometallic catalysis: descriptors, mechanisms and models. AB - With the growing availability of cheap and fast computing resources, increasingly detailed and sophisticated computational studies of homogeneous organometallic catalysis have become feasible. In addition, databases of calculated descriptors can be used to analyse and interpret the key factors contributing to a wide range of experimental observations, facilitating the development of novel catalysts. Such descriptors, most frequently used to capture ligand properties, can also be used to quantify catalyst similarities and derive maps of ligand space which guide experimental screening. Recent publications have shown that descriptor databases can be combined with calculated mechanistic information to gain more detailed insight into catalytic cycles, useful for the design of catalysts. This perspective seeks to describe the contribution of computational studies to the field of homogeneous organometallic catalysis, and to summarise the challenges and opportunities for the future of computational prediction and evaluation of catalytically active transition metal complexes. PMID- 20023962 TI - Recent progress in olefin polymerization catalyzed by transition metal complexes: new catalysts and new reactions. AB - A number of transition metal catalysts including early and late transition metals have been investigated for olefin polymerization and copolymerization. Progress of the field in this decade is reviewed, and important topics among them such as binary catalysts and polymerization of new monomers are described in detail. Our recent results on the polymerization of olefins having cycloalkenes and non conjugated dienes using late transition metal complexes are also mentioned. PMID- 20023963 TI - Probing bioinorganic chemistry processes in the bloodstream to gain new insights into the origin of human diseases. AB - In the context of elucidating the origin of human diseases, past poisoning epidemics have revealed that exceedingly small doses of inorganic environmental pollutants can result in dramatic effects on human health. Today, numerous organic and inorganic pollutants have been quantified in human blood, but the interpretation of these concentrations remains--from a public health point of view--problematic. Conversely, the biomolecular origin for several grievous human diseases is essentially unknown. Taken together and viewed in the context of recent bioinorganic research findings, the established human blood concentrations of toxic metals and metalloids may be functionally connected with the etiology of specific human diseases. To unravel the underlying biomolecular mechanisms, and taking into account the basic flow of dietary matter through mammalian organisms, a better understanding of the bioinorganic chemistry of toxic metals and metalloid compounds in the bloodstream is emerging as a promising avenue for future research. To this end, the concerted application of modern proteomic methodologies, synchrotron-based X-ray absorption spectroscopy and established spectroscopic techniques will contribute to better define the role that blood based bioinorganic chemistry-related processes play in the origin of human diseases. The application of this and other modern proteomic methodologies could contribute to a better understanding of the role that blood-based bioinorganic chemistry-related processes play in the origin and etiology of human diseases. PMID- 20023964 TI - Modern multiphase catalysis: new developments in the separation of homogeneous catalysts. AB - Homogeneous catalysts are powerful tools for the synthesis of fine chemicals, pharmaceuticals and materials, however their exploitation on an industrial scale is often held back due to the challenges of separating and recycling the catalyst. This perspective focuses on approaches to multiphase catalysis that have emerged in the last decade, highlighting methods that can address the separation issues and in some cases result in superior catalyst performance and environmental benefits. PMID- 20023965 TI - Probing the kinetics of ligand exchange on colloidal gold nanoparticles by surface-enhanced Raman scattering. AB - We report a new approach to study the kinetics of ligand exchange on colloidal gold nanoparticles, where the coordination and dissociation of surface ligands could be monitored in situ by surface-enhanced Raman scattering. PMID- 20023966 TI - U(IV) and U(V) azide complexes supported by amide or aryloxide ligands. AB - The uranium(IV) azides [Li(THF)(3)](2)[U(OAr)(4)(N(3))(2)], Ar = 2,6 Me(2)C(6)H(3), and {[Na(THF)(4)][U[N(SiMe(3))(2)](3)(N(3))(2)]}(x) have been synthesised and structurally characterised. Oxidation of these complexes affords [Li(THF)(3)][U(OAr)(5)(N(3))] and U[N(SiMe(3))(2)](3)(N(3))(2), which are the first azides of U(V). PMID- 20023967 TI - Host-guest influence on metallo-supramolecular assemblies with a cyclotriveratrylene-type ligand. AB - The structure-directing effect of guest binding in metallo-supramolecular assemblies of host-type ligands is observed with a 2-D coordination polymer formed between Cd(OAc)(2) and tris(4-pyridylmethylamino)cyclotriguaiacylene in the presence of the globular guest o-carborane, and a trinuclear Cd(II) complex formed in solution in the absence of the bulky guest. PMID- 20023968 TI - Facile entry to 3d late transition metal boryl complexes. AB - Mononuclear nickel and cobalt boryl complexes have been prepared via sigma-bond metathesis reactions and in the case of nickel, an intermediate comprised of a Lewis acid-base type adduct has been experimentally detected by (31)P NMR spectroscopy and its structure probed by DFT calculations. PMID- 20023969 TI - Zirconium bis(pyridonate): a modified amidate complex for enhanced substrate scope in aminoalkene cyclohydroamination. AB - A new bis(amidate)zirconium bis(amido) hydroamination pre-catalyst using 6-tert butyl-3-phenyl-2-pyridone as a proligand has been prepared and characterized. This rare example of an early transition metal complex incorporating a 2 pyridonate derivative as an ancillary ligand was found to be effective for the cyclohydroamination of aminoalkenes, including more challenging substrates bearing unactivated internal C=C bonds. PMID- 20023970 TI - Mono-alkylated bisphosphines as dopants for ESI-MS analysis of catalytic reactions. AB - Bisphosphines Ph(2)P(CH(2))(n)PPh(2) (n = 1, 2, 4, 6) may be easily monobenzylated to generate cationic phosphine/phosphonium ligands [Ph(2)P(CH(2))(n)PPh(2)CH(2)Ph](+). These ligands may be doped into a catalytic reaction involving neutral complexes with labile phosphine ligands, and replacement of a neutral phosphine with a charged analogue renders the resulting complex amenable to electrospray ionisation mass spectrometry (ESI-MS). Examination of olefin hydrogenation with Wilkinson's catalyst, RhCl(PPh(3))(3), revealed that this approach yielded rapid identification of all off-cycle solution species as well as catalyst poisons. Reactive intermediates could be generated using collision-induced dissociation (CID) of a triphenylphosphine ligand to make three-coordinate RhClP(2) species, and these react with alkenes in the gas phase to form RhClP(2)(alkene). The solution speciation and gas phase behaviour revealed by ESI-MS match closely to what is already known about the system from kinetic and NMR studies. PMID- 20023971 TI - Three-coordinate late transition metal fluorinated alkoxide complexes. AB - Homoleptic fluorinated alkoxide complexes have been prepared from KOC4F9, 1, via salt metathesis routes. One four-coordinate K{K(18C6)}[Co(OC4F9)4], 2, and four three-coordinate complexes: {K(18C6)}[Fe(OC4F9)3], 3, {K(18C6)}[Co(OC4F9)3], 4, {K(18C6)}[Cu(OC4F9)3], 5, and {K(18C6)}[Zn(OC4F9)3], 6, have been prepared and all except 5 have been characterized crystallographically. Compounds 3, 4, and 6 are very rare examples of monomeric, trigonal alkoxide complexes. All compounds have been characterized with UV-vis and IR spectroscopy, solution magnetic susceptibility, and elemental analysis. In solution, compound 2 exists in an equilibrium with 4 and 1, which has been probed with cyclic voltammetry, supporting energetically different Co2+/Co3+ potentials in the three-coordinate (E(p,a) = approximately 1.2 V vs Fc/Fc+) and four-coordinate (E(p,a) = approximately 0.9 V) geometries. The ligand field engendered by the perfluoro-t butoxide ligand has been studied with DFT calculations on 4 and the hypothetical [Co(OC4H9)3]- as well as the previously reported [Co(mes)3]- and [Co{N(TMS)2}3]- showing significant -type interactions in the xy plane as well as above and below for the two alkoxide species. PMID- 20023972 TI - Versatile assembly of p-carboxylatocalix[4]arene-O-alkyl ethers. AB - Crystallisation of lower-rim tetra-O-alkylated p-carboxylatocalix[4]arenes from pyridine results in the formation of both bi-layer and pillar type supramolecular motifs. Full alkylation at the calixarene lower rim has significant influence over the supramolecular self-assembly motif, including preclusion of pyridine guest molecules from the calixarene cavity in the solid state. PMID- 20023973 TI - Using metal-ligand interactions for the synthesis of metallostar polymers. AB - In this work we report the synthesis and assembly of metallostar polymers using a combination of metal complexation, RAFT and Click chemistry. PMID- 20023974 TI - A new hybrid scorpionate ligand: a study of the metal-boron bond within metallaboratrane complexes. AB - A new boron-based hybrid scorpionate ligand based upon one 2-mercaptopyridine (mp) and two 1-methyl-imidazoyl-2-thione (mt) units, Na[HB(mt)(2)(mp)] has been prepared. This new ligand together with the recently reported ligand K[HB(mp)(3)] have been used to prepare analogues of the original metallaboratrane complex [Ru(CO)(PPh(3)){kappa(4)-SSBS-B(mt)(3)}]. The effect of tautomerisation of the ligand arms upon the electronic properties of the boron and metal centres is examined in an attempt to probe further the nature of the metal-boron bond within metallaboratrane complexes. PMID- 20023975 TI - Chelating tris(amidate) ligands: versatile scaffolds for nickel(II). AB - The synthesis and characterization of nickel complexes supported by a family of open-chain, tetradentate, tris(amidate) ligands, [N(o-PhNC(O)R)(3)](3-) ([L(R)](3 ) where R = (i)Pr, (t)Bu, and Ph) is described. The complexes [Ni(L(iPr))](-), [Ni(L(tBu))](-), and [Ni(L(Ph))(CH(3)CN)](-) have been characterized by solution state spectroscopic methods and single crystal X-ray diffraction. Each ligand gives rise to a different primary coordination sphere about the nickel centre. These studies indicate that the ligands' acyl substituents can be used to regulate the coordination mode of the amidate donors to nickel and the coordination number of the nickel centres. In addition, the ability of these complexes to bind cyanide has been explored. These experiments demonstrate that only one of these complexes, [Ni(L(iPr))](-), is able to irreversibly bind cyanide and can be used to assemble [Et(4)N](3)[Ni(L(iPr))(mu(2)-CN)Co(L(iPr))], a cyanide bridged, heterobimetallic complex. The synthesis and characterization of the cyanide containing complexes, including magnetic susceptibility studies, are described. PMID- 20023976 TI - Spontaneous formation of crystalline lithium molybdate from solid reagents at room temperature. AB - Lithium molybdate has been prepared by grinding LiOH x H(2)O with MoO(3) in air at room temperature. X-Ray powder diffraction data show that the formation of highly crystalline Li(2)MoO(4) is largely complete after 10 min. The phenacite structure of this material is the same as that derived from an X-ray diffraction study of a single crystal obtained from aqueous solution [R3; a = 14.3178(14) A, c = 9.5757(9) A]. Anhydrous lithium hydroxide fails to give the same reaction indicating that the water of crystallisation of LiOH x H(2)O is a vital component in this rapid synthesis. Differential scanning calorimetry measurements show that this reaction can proceed spontaneously between the two stable solid reagents at sub-ambient temperatures and is driven by the liberation of water from the crystalline lattice. Lithium molybdate prepared in this manner has significantly smaller and more regularly shaped particles than samples prepared by other synthetic methods. PMID- 20023977 TI - Tautomerisation and hydrogen-bonding interactions in four-coordinate metal halide and azide complexes of N-donor-extended dipyrromethanes. AB - The synthesis and structures of Fe, Co, and Zn halide complexes [MX(2)(H(2)L)] (M = Fe, X = Br; M = Co, Zn, X = Cl) of the N-donor extended dipyrromethane ligand H(2)L are described, from which it is clear that bond rearrangements from imine pyrrole to amine-azafulvene tautomers occur on metal co-ordination, both in the solid state and in solution. In the structure of [FeBr(2)(H(2)L)], this H migration results in a pendant amine that is involved in both inter- and intramolecular hydrogen bonds to the bromide ligands, so forming a dimer. As the tautomerisation renders the N-H protons less acidic, metal-based ligand substitution reactions can occur in favour of deprotonation. As such, the reaction between [MCl(2)(H(2)L)] (M = Co, Zn) and NaN(3) results in the formation of the bis(azide) complexes [M(N(3))(2)(H(2)L)] which for Co displays both inter- and intramolecular N-H...N(3)-Co hydrogen bonds in the solid state. In contrast, reactions of the dihalides with the lithium bases LiNMe(2) or LiMe (M = Fe), or reduction reactions with C(8)K (M = Fe, Co) result in the formation of the known dinuclear helicates [M(2)(L)(2)]. PMID- 20023978 TI - Reactivity studies of group 15 Zintl ions towards homoleptic post-transition metal organometallics: a 'bottom-up' approach to bimetallic molecular clusters. AB - Reactions between ethylenediamine (en) solutions of the intermetallic Zintl phases K(3)E(7) (E = P, As) and a series of homoleptic post-transition metal organometallics such as Cu(5)(Mes)(5), M(C(6)H(5))(2) (M = Zn, Cd) and In(C(6)H(5))(3) have yielded a family of novel bimetallic cluster anions. These new species were isolated as [K(2, 2,2-crypt)](+) salts in [K(2,2,2 crypt)](4)Cu(2)E(14) (E = P (1), As (2)), [K(2,2,2-crypt)](4)ZnE(14) (E = P (3), As (4)), [K(2,2,2-crypt)](4)CdP(14) x 6py (5) and [K(2,2,2-crypt)](2)E(7)InPh(2) (E = P (6), As (7)). Species 2, 3, 5 and 6 were crystallographically characterised by single-crystal X-ray diffraction. The stability of all of the cluster anions in solution was confirmed by electrospray mass-spectrometry and by (1)H, (13)C{(1)H} and (31)P{(1)H} NMR spectroscopy when possible. PMID- 20023979 TI - Chromophoric dipyrrin complexes capable of binding to TiO2: synthesis, structure and spectroscopy. AB - The synthesis, characterisation, and TiO2 binding studies of a series of chromophoric complexes of 5-(4-carboxyphenyl)-4,6-dipyrrin (L(b)) are presented. The synthesis of [Ru(bipy)(L(b))2] (bipy = 2,2-bipyridine), [Rh(L(b))3], and [Pd(L(b))2] was achieved by initial coordination of 5-(4-methoxycarbonylphenyl) 4,6-dipyrrin (L(a)) followed by hydrolysis of the ester group. The carboxyl groups that are located on the peripheries of these complexes are able to engage in intermolecular hydrogen bonding interactions in the solid state, as revealed by X-ray crystallography. These groups also allow the complexes to anchor to the surface of TiO2 nanoparticles, as evidenced by colouration of the TiO2 and FT-IR spectroscopy. The ability of these complexes to capture a significant fraction of sunlight and to adhere to TiO2 surfaces renders them viable dyes for photochemical devices such as dye sensitised solar cells. PMID- 20023980 TI - Is there really a diagnostically useful relationship between the carbon-oxygen stretching frequencies in metal carboxylate complexes and their coordination mode? AB - An investigation into the relationship between the asymmetric and symmetric stretching modes of ancillary carboxylate ligands and their mode of binding in a family of manganese(III) complexes of tetradentate N(2)O(2) Schiff base ligands, which includes comparison of an (18)O isotopically labelled derivative, has been undertaken. The results suggest that caution should be employed in using this technique to assign the binding mode of the carboxylate ligands in these complexes despite its very extensive use in the literature. PMID- 20023981 TI - Insertion of platinum and palladium into a sulfur(IV)-sulfur(II) bond of a sulfur substituted sulfurane. AB - Transition metal complexes are known to react with disulfides and insert into the disulfide bond, but there has been no report on similar reactivity towards the hypervalent sulfur(IV)-sulfur(II) bond compound, a sulfur-substituted sulfurane, because of its instability. In this paper, we report the reactions of the sulfur substituted sulfurane 1 with platinum and palladium complexes, in which the corresponding complexes 2 and 3 with an O,S,S'-ligand were obtained by bond cleavage of both the sulfur(IV)-sulfur(II) bond and the sulfur(IV)-oxygen bond. The structures of the complexes were characterized by NMR spectroscopic studies, elemental analysis, and X-ray crystallographic analysis. The palladium complex 3 underwent elimination of triphenylethylene episulfide when treated with dppe to give the dppe palladium complex 4 with an O,S-ligand, which was also obtained by the reaction of Pd(dba)2 and dppe with the sulfurane 1. X-ray crystallographic analysis of complexes 2 and 4 revealed a square-planar structure around the platinum or palladium. The formation mechanisms of these complexes 2-4 are discussed. PMID- 20023982 TI - The interaction of rare earth chlorides with 4,4'-bipyridine for the reversible formation of template based luminescent Ln-N-MOFs. AB - By solvent free syntheses of the rare earth trichlorides LnCl(3), Ln = Pr, Nd, Sm, Eu, Tb with melts of 4,4'-bipyridine two-dimensional frameworks of the formula (2)(infinity)[Ln(2)Cl(6)(4,4'-bipy)(3)] x 2(4,4'-bipy) are obtained, with 4,4'-bipy = C(10)H(8)N(2), 4,4'-bipyridine. 4,4'-Bipyridine acts both as a linker ligand as well as a template and populates all cavities in the structures. The template can be evaporated at temperatures >200 degrees C giving a new high temperature compound with a yet unknown structure. Further heating results in the release of the remaining linking equivalents of the ligand and reformation of LnCl(3). Thus the reaction can be run in cycles if the evaporated ligand is collected. Luminescence in the visible range without quenching by concentration is observed for the lanthanides Eu and Tb, which is identified for Eu(III) with the transitions (5)D(0)-->(7)F(J) and (5)D(4)-->(7)F(J) for Tb(III). The hybrid character of the material is reflected by the additional strong antenna effect of the ligand: main excitation is observed via the 4,4'-bipyridine linkers followed by an energy transfer to the metal centres. It is remarkable that both the template containing MOFs as well as the new high temperature compounds exhibit luminescence properties. PMID- 20023983 TI - A copper-activated magnetic resonance imaging contrast agent with improved turn on relaxivity response and anion compatibility. AB - We present the synthesis and characterization of Copper-Gad-7 (CG7), a new copper activated magnetic resonance imaging (MRI) contrast agent that possesses a Gd(3+) DO3A scaffold with an appended thioether-rich receptor for copper recognition. Installation of additional carboxylate groups on the periphery of the CG scaffold affords a practical strategy to increase the absolute relaxivity of these types of copper-responsive MRI sensors as well as reduce their sensitivity to biologically abundant anions. Due in large part to restricted access of inner sphere water molecules to the paramagnetic Gd(3+) core, in the absence of copper ions, CG7 exhibits a relatively low relaxivity value of r(1) = 2.6 mM(-1) s(-1); addition of Cu(+) triggers a 340% enhancement in relaxivity to r(1) = 11.4 mM(-1) s(-1). For comparison, the relaxivity of the analogous CG2 sensor without peripheral carboxylates increases from r(1) = 1.5 to 6.9 mM(-1) s(-1) upon Cu(+) binding. CG7 features high selectivity for Cu(+) over a range of biologically relevant metal ions, including the cellular abundant alkali and alkaline earth cations and d-block ions Zn(2+) and Cu(2+). Moreover, the Cu(+)-response of the CG7 sensor is not significantly affected by bicarbonate, phosphate, citrate, and lactate anions at cellular levels. (17)O NMR dysprosium-induced shift (DIS) and nuclear magnetic relaxation dispersion (NMRD) experiments suggest that the origin of the improved anion compatibility of CG7 is a reduced q modulation compared to previous members of the CG family, and T(1)-weighted phantom images confirm that CG7 can monitor changes in copper levels by MRI at clinically relevant field strengths. PMID- 20023984 TI - Reactivity and kinetic-mechanistic studies of regioselective reactions of rhodium porphyrins with unactivated olefins in water that form beta-hydroxyalkyl complexes and conversion to ketones and epoxides. AB - This article reports on the selective oxidation of unactivated alkenes to ketones and epoxides through the intermediacy of beta-hydroxyalkyl rhodium porphyrin complexes which are formed by reactions of terminal alkenes with tetra(p sulfonatophenyl)porphyrin rhodium(III) complex. The beta-hydroxyalkyl rhodium porphyrin complexes in water undergo beta-C-H elimination to produce ketones in aqueous pH 9.0 solutions and O-H deprotonation in KOH/DMSO solutions resulting in the rapid and quantitative intramolecular nucleophilic displacement to form 1,2 epoxyalkanes. PMID- 20023985 TI - Syntheses and structures of zirconium(IV) complexes supported by 2,6-di adamantylaryloxide ligands and formation of arene-bridged dizirconium complexes with an inverse sandwich structure. AB - The reaction of 2,6-di-adamantyl-4-R-phenol (Ar(R)OH, R = Me, tBu) with nBuLi in THF gave good yields of the lithiated derivatives (Ar(R)O)Li(THF)2. Addition of 2 equiv. of (ArRO)Li(THF)2 to ZrCl4(THF)2 in THF afforded the base-free dichloride complexes (Ar(R)O)ZrCl2 (R = Me 1a, tBu 1b). The dibenzyl derivative (Ar(Me)O)2Zr(CH2Ph)2 2 was synthesized by addition of 2 equiv. of PhCH2MgCl to 1a in toluene. Reduction of the dichloride complexes 1a and 1b with KC8 in toluene led to the formation of the toluene-bridged dizirconium complexes [(Ar(R)O)2Zr]2( 6:6-C7H8) (R = Me 3a, tBu 3b). A similar reaction was carried out in benzene to produce the corresponding benzene-bridged analogue [(Ar(R)O)2Zr]2(mu eta(6):eta(6)-C6H6) (R = Me 4a, tBu 4b). Treatment of 3a with 2 equiv. of 1 azidoadamantane (AdN3) in THF resulted in the release of toluene and N2, generating the monomeric imide complex (Ar(Me)O)2Zr(NAd)(thf) 5. When the analogous reaction was carried out in toluene, (Ar(Me)O)2Zr(AdNN=NNAd) 6 was obtained. Structures of (Ar(R)O)Li(THF)2, 1a, 2 and 5 were determined by X-ray crystallography. PMID- 20023986 TI - Phosphorus stabilized carbene complexes: bisphosphonate dianion synthesis, reactivity and DFT studies of O~C~O zirconium(IV) complexes. AB - The reactivity of the geminal dianion of tetraisopropyl methylenediphosphonate, 3, with [ZrCl4(THF)2] affords a trinuclear structure 5 in which an unprecedented triscarbene-Zr dianionic fragment is found. The overall trinuclear arrangement is assembled by PO bridging moieties. The X-ray crystal structure of 5 is presented. The reaction of complex 5 with stoichiometric amounts of pyridine affords a dinuclear complex 6 whose formulation was ascertained by an X-ray crystal structure. Complexes 5 and 6 cleanly react with aldehydes (R1)(H)CO to afford the corresponding olefin of general formula [((OiPr)2P=O)2C=C(R1)(H)]. DFT calculations which were carried out on the model complex of 5t and a hypothetical monometallic Zr complex [Zr(3)Cl2(OMe2)2] 8t indicate that the Zr-C bond only features a weak pi-interaction between the carbon atom and a vacant orbital at the metal. The polarization of the orbital towards the carbon center is consistent with the observed nucleophilic character of the carbene complexes described here. PMID- 20023987 TI - Lanthanide tri-benzyl complexes: structural variations and useful precursors to phosphorus-stabilised lanthanide carbenes. AB - Reaction of [Ln(I)3(THF)4] (Ln = Ce, Pr) or [Ln(I)3(THF)3.5] (Ln = Nd, Sm, Gd, Dy, Er) with three equivalents of [KBz] (Bz = CH2C6H5) at 0 degrees C afforded the corresponding lanthanide tri-benzyl complexes [Ln(Bz)3(THF)3] [Ln = Ce (2), Pr (3), Nd (4), Sm (5), Gd (6), Dy (7), Er (8) La (11)] in 48-75% crystalline yields, with the exception of the redox active samarium complex, which was isolated in poor (20%) yield. Complexes 2-8 were found to adopt distorted octahedral geometries, where the Bz and THF groups are bound in a mutually fac manner in the solid state. Although the series is structurally similar, classification of three structural types can be made on the basis of the lanthanide contraction: (i) complexes which exhibit three eta(2) Ln...C(ipso) contacts (1-4, 11); (ii) complexes which show one eta(2) Ln...C(ipso) contact (5); (iii) complexes with no multi-hapto interactions (6-8). For ytterbium, the mixed valence, Yb(II)/Yb(III) complex [Yb(II)(Bz)(THF)5]+[Yb(III)(Bz)4(THF)2]- (9) was reproducibly formed at 0 degrees C and -78 degrees C as a result of partial (50%) Yb(III) --> Yb(II) reduction with concomitant formation of half an equivalent of 1,2-diphenylethane by oxidative coupling. Tri-valent [Yb(Bz)3(THF)3] (10) was apparently not formed. The synthetic utility of tri benzyl lanthanide complexes 2-8 and 11 were tested in reactions with the bis (iminophosphorano)methane H2C(PPh2NSiMe3)2 (H2-BIPM), which afforded [Ln(BIPM)(H BIPM)] [Ln = La (12), Ce (13), Pr (14), Nd (15), Sm (16), Gd (17)] and [Ln(BIPM)(Bz)(THF)] [Ln = Dy (18), Er (19)]. Compounds 2-9 and 12-19 have been variously characterised by X-ray crystallography, multi-nuclear NMR spectroscopy, FTIR spectroscopy, room temperature Evans method solution magnetic moments and CHN micro-analyses. PMID- 20023988 TI - cis-2,6-Dimethylpiperidide: a structural mimic for TMP (2,2,6,6 tetramethylpiperidide) or DA (diisopropylamide)? AB - Four novel heterobimetallic ate complexes containing cis-2,6-dimethylpiperidide (cis-DMP) have been prepared and characterised. Two contain one cis-DMP ligand, namely the bisalkyl-amido lithium, and sodium zincates [(TMEDA) x MZn(cis DMP)(tBu)2] (M = Li for 1, Na for 2). Both 1 and 2 are synthesised by co complexation of the respective alkali metal amide with di-tert-butylzinc in the presence of a molar equivalent of N,N,N',N'-tetramethylethylenediamine (TMEDA) in a hydrocarbon medium. The third complex, containing two cis-DMP ligands, is the alkyl-bisamido sodium zincate [(TMEDA) x NaZn(cis-DMP)2(tBu)] 3. Complex 3 is prepared from 2 via a disproportionation reaction where the by-product is [(TMEDA) x NaZn(tBu)3]. Another alkyl-diamido sodium zincate, [(TMEDA) x NaZn(DIBA)2(tBu)] 4 is synthesised by utilising diisobutylamine [DIBA(H)]. This reaction emphasises the generality of this disproportionation process. Complex 5 contains three cis-DMP ligands and is a tris-amido sodium magnesiate [(TMEDA) x NaMg(cis-DMP)3]. It is prepared by treating an equimolar mixture of butylsodium and dibutylmagnesium with three and one molar equivalents of cis-DMP(H) and TMEDA respectively, in hydrocarbon solution. By comparison of 1-5 with appropriate complexes from the literature, it has been possible to experimentally determine that the steric bulk of cis-DMP closely resembles that of DA but is considerably less bulky than 2,2,6,6-tetramethylpiperidide (TMP). PMID- 20023989 TI - New insights into addition reactions of dialkylzinc reagents to trifluoromethyl ketones: structural authentication of a beta-hydride elimination product containing a tetranuclear zinc chain. AB - A systematic study of the stoichiometric alkylation reactions of 2,2,2 trifluoroacetophenone 1 with [ZnR(2)(TMEDA)] (R= Me, Et, (t)Bu, CH(2)SiMe(3); TMEDA= N,N,N',N'-tetramethylethylenediamine) monitored by (1)H and (19)F NMR spectroscopy is presented. For R = Me, Et the alkylation products alkyl(alkoxides) [(TMEDA)Zn(R){OC(CF(3))(R)Ph}] (R = Me, 2: Et, 3) are obtained as the single products of the reaction. When the steric bulk of the dialkylzinc reagent is increased the alkylation reaction is inhibited. Thus, for R = (t)Bu, the reduction product [(TMEDA)Zn((t)Bu){OC(CF(3))(H)Ph}] is obtained as a result of beta-hydride elimination from one of the (t)Bu groups of the organometallic reagent. (1)H NMR spectroscopic monitoring of the reaction allowed the detection of isobutene as a side product of this reduction process. For the highly sterically demanding group R = CH(2)SiMe(3) which lacks hydrogen atoms at the beta position, no reaction is observed even under refluxing conditions. Two important intermediates from these reactions have been structurally elucidated: [(TMEDA)Zn(Me){OC(CF(3))(Me)Ph}] (2) which could be involved in the previously reported alkylation reaction of trifluoromethyl ketones by ZnR(2) catalysed by TMEDA and unprecedented tetranuclear [((t)Bu)(2)Zn(4){OC(CF(3))(H)Ph}(6)] (5) resulting from the reduction of 1 when reacted with (t)Bu(2)Zn, which displays a rare Zn...Zn...Zn...Zn linear chain arrangement for a zinc alkyl(alkoxide). PMID- 20023990 TI - A new hexakis(isocyanato)silicate(IV) and the first neutral Lewis-base adducts of silicon tetraisocyanate. AB - The new silicates K2Si(NCE)6, E = O, S (1a,b) were synthesised directly by reaction of silicon tetrachloride with potassium cyanate or thiocyanate. 1a,b can be converted to the salts (PPN)2[Si(NCE)6] (2a,b) with bulky cations (PPN+ = N(PPh3)2+), which contain isolated silicate dianions. Reaction of 1a with diimines affords the hexacoordinate complexes Si(NCO)4L, L = bpy (3), phen (4), which are the first isolated neutral adducts to Si(NCO)4. The compounds were characterised by a combination of IR and NMR spectroscopies, MS, TGA, DSC and X ray diffraction. PMID- 20023991 TI - Synthesis and structural characterization of well-defined anionic aluminum alkoxide complexes supported by NON-type diamido ether tridentate ligands and their use for the controlled ROP of lactide. AB - The tridentate proligands (RNH-o-C6H4)2O (1a, R = C5H9; 1b, R = Cy) were found to readily react with LiAlH4 to yield the corresponding lithium aluminium dihydrido salt species [eta(2)-N,N-{(RN-o-C6H4)2O}AlH(mu-H)Li(THF)]2 (2a, R = C5H9; 2b, R = Cy) in 50% and 42% yield, respectively. The solid-state structure of both complexes 2a and 2b were determined by X-ray crystallographic studies. Compounds 2a and 2b readily react with one equivalent of benzaldehyde to afford the corresponding mono-benzyloxide species eta(2)-N,N-{RN-o-C6H4)2O}Al(H)(mu OCH2Ph)Li(THF)2 (4a, R = C4H9; 4b, R = Cy), as confirmed by X-ray studies in the case of 4b. In a similar manner, when compounds 2a and 2b are reacted with two equivalents of benzaldehyde the bis-benzyloxide derivatives 5a and 5b eta(3) N,N,O-{RN-o-C6H4)2O}Al( -OCH2Ph)2Li(THF)2 (5a, R = C4H9; 5b, R = Cy) may be prepared. While the lithium Al mono-alkoxide species 4a-b are inactive in the ring-opening polymerization (ROP) of lactide, their bis-alkoxide Al analogues 5a b polymerize rac-lactide and (S)-lactide at room temperature, which is rather uncommon for Al-based alkoxide systems. Kinetic studies of the lactide ROP initiated by compound 5a suggest a strong preference for racemic enchainment during the ROP chain growth; the resulting PLAs are however moderately heterotactic due to detrimental transesterification processes occurring as the chain grows. PMID- 20023992 TI - Electron rich bidentate phosphinimine-imine ligands: synthesis and reactivity of late transition metal complexes. AB - Electron rich phosphinimine-imine proligands Ph(3)PN(C(6)H(4))C(Ph)(NAr) (L(Ar)) (Ar = 4-(OEt)C(6)H(4) (OEt), 3,5-Me(2)C(6)H(3)(Xyl)) were synthesized in three steps from 2-aminobenzophenone. These compounds, along with previously reported L(Mes) and L(Tol) (Mes = 2,4,6-Me(3)C(6)H(2), Tol = 4-MeC(6)H(4)) were used to synthesize a series of tetracarbonyltungsten(0) complexes: L(Mes)W(CO)(4) (1), L(Tol)W(CO)(4) (2), L(OEt)W(CO)(4) (3), and L(Xyl)W(CO)(4) (4). The ligands were evaluated by analysis of the carbonyl stretching frequencies of the tungsten complexes and were shown to be better sigma-donors and poorer pi-acceptors compared to similar ligands in the literature. The coordination chemistry of the proligands was expanded to other late transition metals and L(Mes)CoCl(2) (5), L(Tol)CoCl(2) (6), L(Mes)NiBr(2) (7), L(Tol)NiBr(2) (8), L(Mes)ZnCl(2) (9), and L(Tol)ZnCl(2) (10) were synthesized by the direct reaction of L(Mes) and L(Tol) with the respective metal dihalide precursors. The complexes were fully characterized and the molecular structures of complexes 3, 6, 7, and 10 were reported. The synthesis of zinc complexes 9 and 10 was dependent on the steric bulk of the ligand. Complex 10 proved to be resistant to derivatization via a number of routes. PMID- 20023993 TI - Structure and magnetic behaviour of mono- and bimetallic chromium(III) complexes of amine-bis(phenolate) ligands. AB - Two lithium amine-bis(phenolate) and four chromium(III) amine-bis(phenolate) complexes have been prepared. The diprotonated tripodal tetradentate ligand precursors 2-tetrahydrofurfuryl-N,N-bis(2-methylene-4-methyl-6-tert-butylphenol), H(2)[O(2)NO](BuMe); 2-tetrahydrofurfuryl-N,N-bis(2-methylene-4,6-tert butylphenol), H(2)[O(2)NO](BuBu); 2-pyridylamino-N,N-bis(2-methylene-4,6 methylphenol), H(2)[O(2)NN'](MeMe); and 2-pyridylamino-N,N-bis(2-methylene-4,6 tert-butylphenol), H(2)[O(2)NN'](BuBu); can be lithiated using n-butyllithium. Isolation of the Li(2)[O(2)NO](RR') compounds generates dimeric {Li(2)[O(2)NO](RR')}(2) (R = t-Bu, R' = Me in 1 and R = R' = t-Bu in 2) in the solid state as shown by single-crystal X-ray diffraction. The lithiated ligands were used to prepare a series of Cr(III) complexes. Monometallic complexes are obtained when prepared and purified under strictly anhydrous conditions, giving CrCl(THF)[O(2)NO](RR') (R = t-Bu, R' = Me in 3 and R = R' = t-Bu in 4). However, bimetallic Cr complexes are obtained upon recrystallization in air, where adventitious water reacts with the complex resulting in protonation of one of the phenolate groups of the ligand and generating hydroxide, which bridges two Cr(III) centres. Solid-state single-crystal X-ray diffraction studies of {CrCl[O(2)NN'](MeMe)}(mu-HO){CrCl[HO(2)NN'](MeMe)}, 5, and {CrCl[O(2)NN'](BuBu)}(mu-HO){CrCl[HO(2)NN'](BuBu)}, 6, were performed. The paramagnetic Cr(III) complexes were also characterized by UV-vis spectroscopy, mass spectrometry and magnetic measurements. PMID- 20023994 TI - Ligand effects in chromium diphosphine catalysed olefin co-trimerisation and diene trimerisation. AB - A series of symmetric and unsymmetric N,N-bis(diarylphosphino)amine ('PNP') ligands (Ar2PN(R)PNAr'2: R = Me, Ar2 = o-anisyl, Ar'2 = Ph, 1, R = Me, Ar2 = o tolyl, Ar'2 = Ph, 2, R = Me, Ar2 = Ph(o-ethyl), Ar'2 = Ph, 3, R = Me, Ar2 = Ar'2 = o-anisyl, 4, R = iPr, Ar2 = Ar'2 = Ph, 5) and symmetric N,N' bis(diarylphosphino)dimethylhydrazine ('PNNP') ligands (Ar2PN(Me)N(Me)PAr2: Ar2 = o-tolyl, 6, Ar2 = o-anisyl, 7) have been synthesised. Catalytic screening for ethene/styrene co-trimerisation and isoprene trimerisation was performed via the in situ complexation to [CrCl3(THF)3] followed by activation with methylaluminoxane (MAO). PNNP catalytic systems showed a significant increase in activity and selectivity over previously reported PNP systems in isoprene trimerisation. Comparing the symmetric and unsymmetric variants in ethene and styrene co-trimerisation resulted in a switch in selectivity, an unsymmetric catalytic (o-anisyl)2PN(Me)PPh2 (1) ligand system affording unique incorporation of two styrenic monomers into the co-trimer product distribution differing from the familiar two ethene and one styrene -substituted alkenes. Complexes of the type [(diphosphine)Cr(CO)4] 8-11 were also synthesised, the single-crystal X-ray diffraction of which are reported. We propose the mechanisms of these catalytic transformations and an insight into the effect of the ligand series on the chromacyclic catalytic intermediates. PMID- 20023995 TI - Toward the development of prochelators as fluorescent probes of copper-mediated oxidative stress. AB - A fluorescent sensor prochelator, FlamB (fluorescein hydrizido 2 imidophenylboronic ester), has been developed that selectively probes for copper under conditions of oxidative stress. High levels of hydrogen peroxide trigger the release of a boronic ester masking group from the prochelator to unveil a metal chelator, FlamS (fluorescein hydrizido 2-imidophenol), that provides a modest fluorescence increase in response to Cu(2+) but not other metal ions. X Ray crystal structures of FlamB, FlamS, and Cu-bound FlamS are all reported. The fluorescence turn-on results from opening of a fluorescein spirolactam ring upon Cu(2+) binding to FlamS in aqueous solution. Oxidation of the aryl boronic ester of FlamB to the metal-binding phenol of FlamS proceeds in organic solvents. However, in aqueous solution a competing mechanism occurs due to hydrolytic instability of the masked prochelator. Hydrolysis of FlamB leads to formation of fluorescein hydrazide, which interacts with copper or H(2)O(2) to produce fluorescein and a significant fluorescence increase. PMID- 20023996 TI - Metathesis catalysts in confining reaction fields--confinement effects vs. surface effects. AB - It has been recognized previously that inside nanosized cavities like in mesoporous materials a small ensemble situation can be realized. The limited number of reactants per confinement leads to unusual chemical behaviour whenever inter- vs. intramolecular pathways compete against each other. Therefore, the metathesis reaction might represent a chemical process for which the product distribution is influenced by a confining reaction field. In the current paper we report about the unusual reactivity of the Grubbs generation I catalyst inside mesoporous silica materials. It has been observed that due to a combination of confinement and surface effects the Grubbs catalyst is not stable inside small pores. It quickly rearranges to a variety of unexpected products. However, the undesired deactivation of the catalyst can be prohibited by surface modification of the silica surfaces. PMID- 20023997 TI - Hydrolytic cleavage of DNA-model substrates promoted by polyoxovanadates. AB - Hydrolysis of 4-nitrophenyl phosphate (NPP) and bis-4-nitrophenyl phosphate (BNPP), two commonly used DNA model substrates, was examined in vanadate solutions by means of (1)H, (31)P and (51)V NMR spectroscopy. The hydrolysis of the phosphoester bond in NPP at 50 degrees C and pH 5.0 proceeds with a rate constant of 1.74 x 10(-5) s(-1). The cleavage of the phosphoester bond in BNPP at 70 degrees C and pH 5.0 proceeds with a rate constant of 3.32 x 10(-6) s(-1), representing an acceleration of four orders of magnitude compared to the uncatalyzed cleavage. Inorganic phosphate and nitrophenol (NP) were the only products of hydrolysis. The NMR spectra did not show evidence of any paramagnetic species, excluding the possibility of V(V) reduction to V(IV), indicating that the cleavage of the phosphoester bond is purely hydrolytic. The pH dependence of k(obs) revealed that the hydrolysis proceeds fastest in solutions of pH 5.5. Comparison of the rate profile with the concentration profile of polyoxovanadates shows a striking overlap of the k(obs) profile with the concentration of decavanadate (V(10)). Kinetic experiments at 37 degrees C using a fixed amount of NPP and increasing amounts of V(10) permitted the calculation of catalytic (k(c) = 5.67 x 10(-6) s(-1)) and formation constants for the NPP-V(10) complex (K(f) = 71.53 M(-1)). Variable temperature (31)P NMR spectra of a reaction mixture revealed broadening and shifting of the (31)P resonance upon addition of increasing amounts of decavanadate and upon increasing temperature, implying the dynamic exchange process between free and bound NPP at higher temperatures. The origin of the hydrolytic activity of V(10) is most likely due its high lability and its dissociation into smaller fragments which may allow the attachment of NPP and BNPP into the polyoxovanadate framework. PMID- 20023998 TI - Olefin metathesis as a tool for multinuclear Co(III)salen catalyst construction: access to cooperative catalysts. AB - The construction of novel (macrocyclic) multinuclear Co(III)salen catalysts is reported. Olefin metathesis has been used as a key construction tool for the multimetallic structures starting from versatile allyl-substituted salen scaffolds. The Co(III) complexes were tested in the hydrolytic kinetic resolution of (rac)-1,2-epoxyhexane and epoxide ring opening reactions using methanol as the nucleophile. The preliminary results suggest a cooperative mode of catalysis in the case of bis-Co(III)salen macrocycle 10. PMID- 20023999 TI - Tuning the phase behavior of ionic liquids in organically functionalized silica ionogels. AB - We have synthesized mesoporous silica monoliths functionalized with 2-(4 pyridylethyl)triethoxysilane 1 and N,N-dimethyl-pyridine-4-yl-(3-triethoxysilyl propyl)-ammonium iodide 2. The organically modified silica monoliths were characterized via IR spectroscopy, nitrogen sorption, small angle X-ray scattering (SAXS), thermogravimetric analysis-differential thermal analysis (TGA DTA), and acid-base titration. The degree of functionalization can be changed by the ratio of the functional silane to the silica precursor tetramethyl orthosilicate (TMOS). The functionalized silica monoliths were filled with 1 ethyl-3-methyl imidazolium [Emim]-X (X = dicyanamide [N(CN)2] or triflate [TfO]) ionic liquids (ILs) using an established methanol-IL exchange technique. The phase behavior of the resulting ionogels was investigated via differential scanning calorimetry (DSC). DSC curves show that the modification of the silica pore walls with organic groups strongly affects the phase behavior of the confined ILs. Modification with silane 1 completely suppresses the glassy state of [Emim][TfO] previously observed in unmodified silica monoliths (Gobel et al., Phys. Chem. Chem. Phys. 2009, 11, 3653). In contrast, modification with silane 2 leads to the appearance and disappearance, respectively, of a presumed additional phase in [Emim][TfO] and [Emim][N(CN)2] with varying degree of monolith functionalization. The data thus show that organic modification of silica matrix materials could be a viable approach for the tuning of ionogel properties. PMID- 20024000 TI - A multicomponent CuAAC "click" approach to a library of hybrid polydentate 2 pyridyl-1,2,3-triazole ligands: new building blocks for the generation of metallosupramolecular architectures. AB - A one pot, multicomponent CuAAC reaction has been exploited for the safe generation of alkyl, benzyl or aryl linked polydentate pyridyl-1,2,3-triazole ligands from their corresponding halides, sodium azide and alkynes in excellent yields. The ligands have been fully characterised by elemental analysis, HR-ESMS, IR, (1)H and (13)C NMR and in two cases the structures were confirmed by X-ray crystallography. Additionally, we have examined the Ag(I) coordination chemistry of these ligands and found, using HR-ESMS, (1)H NMR, and X-ray crystallography, that both discrete and polymeric metallosupramolecular architectures can be formed. PMID- 20024001 TI - Acetylenedithiolate as directional bridging ligand in cobalt(I) alkyne platinum dithiolato bimetallic complexes. AB - The eta(2)-C,C-acetylenedithiolate (acdt2-) complex K[(triphos)Co(acdt)], K-5, {triphos = 1,1,1-tris(diphenylphosphinomethyl)ethane} was obtained by consecutive removal of S-protection groups in [(triphos)Co(1)]PF6, 3-PF6 (1 = 1 Trimethylsilyl-7-phenyl-3,6-dithiahept-4-ine). Reaction of K-5 with selected Pt(II) salts resulted in the formation of the new heterobimetallic complexes [(phen)Pt(5)]BPh4, 6-BPh4, (phen = 1,10-phenanthroline) and [(dppe)Pt(5)]BPh4, 7 BPh4, {dppe = 1,2-bis(diphenylphosphino)ethane}, which were fully characterized. X-ray diffraction studies showed that Co and Pt are linked by acdt2- in the eta(2)-C,C-2-S,S-bridging mode. The electronic structure of 6-BPh4 and 7-BPh4 was investigated by electronic absorption spectroscopy, cyclic voltammetry and X-band EPR spectroscopy of neutral 7. In addition, NMR spectroscopy, X-ray diffraction and reactivity studies with the alkyne complexes [(PMe3)3Co(1)]-PF6, 2-PF6, with 3-PF6 and the intermediate product [(triphos)Co{eta(2)-(S)C2(SCH2Ph)}], 4, uncovered the flexibility of the CoC2S2-moiety within the persisting complex scaffold throughout the synthetic scheme. PMID- 20024003 TI - Bis(oxazolinyl)phenylborane: a Lewis acid-containing ligand for methide abstraction-based coordination to aluminum(III). AB - A compound that contains a Lewis acidic boron center and coordinating oxazoline groups, bis(4,4-dimethyl-2-oxazolinyl)phenylborane (PhB(Ox(Me2))2; 1), has been prepared and spectroscopically characterized. Solvent dependent 15N and 11B NMR spectroscopic properties and solid-state 11B NMR measurements provide support for intermolecular interactions involving Lewis acid and base sites. The bifunctional nature of oxazolinylborane 1 is demonstrated by its reaction with (AlMe3)2, which proceeds via methide abstraction by the boron and oxazoline coordination to aluminum to yield [(kappa(2)-PhMeB(Ox(Me2))2AlMe2] (2). Compound 2 contains a planar six-membered chelate ring, in contrast to related bis(pyrazolyl)boratoaluminum compounds that are puckered. Additionally, compound 2 and related bidentate tris(oxazolinyl)phenylborato dimethylaluminum are inert toward aluminum-methyl bond protonolysis. This robust nature suggested the possibility of using these oxazolinylboratoaluminum compounds in catalytic reactions, as is demonstrated by lactide ring-opening polymerization. PMID- 20024002 TI - Synthesis and reactivity of palladium(II) fluoride complexes containing nitrogen donor ligands. AB - This article describes the synthesis, characterization, and reactivity of palladium(II) fluoride complexes containing sp(2) and sp(3) nitrogen-containing supporting ligands. Both cis and trans complexes of general structure (N)(N')Pd(II)(R)(F) (R = Ar or CH(3)) as well as cis-(N)(2)Pd(II)(F)(2) are reported. Crystallographic characterization of these molecules has allowed structural comparisons to related phosphine-ligated species. Furthermore, these studies have revealed that nitrogen-donor ligands support some of the longest and the shortest Pd-F bonds reported to date. The thermal decomposition of (N)(N')Pd(II)(R)(F) has also been examined, and no products of C-F bond-forming reductive elimination were obtained in any case. PMID- 20024007 TI - The Brooklyn traffic real-time ambient pollutant penetration and environmental dispersion (B-TRAPPED) study. PMID- 20024008 TI - Overview of the Brooklyn traffic real-time ambient pollutant penetration and environmental dispersion (B-TRAPPED) study: theoretical background and model for design of field experiments. AB - The Brooklyn traffic real-time ambient pollutant penetration and environmental dispersion (B-TRAPPED) study was a multidisciplinary field research project that investigated the transport, dispersion, and infiltration processes of traffic emission particulate matter (PM) pollutants in a near-highway urban residential area. The urban PM transport, dispersion, and infiltration processes were described mathematically in a theoretical model that was constructed to develop the experimental objectives of the B-TRAPPED study. In the study, simultaneous and continuous time-series PM concentration and meteorological data collected at multiple outdoor and indoor monitoring locations were used to characterize both temporal and spatial patterns of the PM concentration movements within microscale distances (<500 m) from the highway. Objectives of the study included (1) characterizing the temporal and spatial PM concentration fluctuation and distribution patterns in the urban street canyon; (2) investigating the effects of urban structures such as a tall building or an intersection on the transport and dispersion of PM; (3) studying the influence of meteorological variables on the transport, dispersion, and infiltration processes; (4) characterizing the relationships between the building parameters and the infiltration mechanisms; (5) establishing a cause-and-effect relationship between outdoor-released PM and indoor PM concentrations and identifying the dominant mechanisms involved in the infiltration process; (6) evaluating the effectiveness of a shelter-in-place area for protection against outdoor-released PM pollutants; and (7) understanding the predominant airflow and pollutant dispersion patterns within the neighborhood using wind tunnel and CFD simulations. The 10 papers in this first set of papers presenting the results from the B-TRAPPED study address these objectives. This paper describes the theoretical background and models representing the interrelated processes of transport, dispersion, and infiltration. The theoretical solution for the relationship between the time-dependent indoor PM concentration and the initial PM concentration at the outdoor source was obtained. The theoretical models and solutions helped us to identify important parameters in the processes of transport, dispersion, and infiltration. The B TRAPPED study field experiments were then designed to investigate these parameters in the hope of better understanding urban PM pollutant behaviors. PMID- 20024009 TI - The Brooklyn traffic real-time ambient pollutant penetration and environmental dispersion (B-TRAPPED) field study methodology. AB - The Brooklyn Traffic Real-Time Ambient Pollutant Penetration and Environmental Dispersion (B-TRAPPED) field study examined indoor and outdoor exposure to traffic-generated air pollution by studying the individual processes of generation of traffic emissions, transport and dispersion of air contaminants along a roadway, and infiltration of the contaminants into a residence. Real-time instrumentation was used to obtain highly resolved time-series concentration profiles for a number of air pollutants. The B-TRAPPED field study was conducted in the residential Sunset Park neighborhood of Brooklyn, NY, USA, in May 2005. The neighborhood contained the Gowanus Expressway (Interstate 278), a major arterial road (4(th) Avenue), and residential side streets running perpendicular to the Gowanus Expressway and 4(th) Avenue. Synchronized measurements were obtained inside a test house, just outside the test house facade, and along the urban residential street canyon on which the house was located. A trailer containing Federal Reference Method (FRM) and real-time monitors was located next to the Gowanus Expressway to assess the source. Ultrafine particulate matter (PM), PM(2.5), nitrogen oxides (NO(x)), sulfur dioxide (SO(2)), carbon monoxide (CO), carbon dioxide (CO(2)), temperature, relative humidity, and wind speed and direction were monitored. Different sampling schemes were devised to focus on dispersion along the street canyon or infiltration into the test house. Results were obtained for ultrafine PM, PM(2.5), criteria gases, and wind conditions from sampling schemes focused on street canyon dispersion and infiltration. For comparison, the ultrafine PM and PM(2.5) results were compared with an existing data set from the Los Angeles area, and the criteria gas data were compared with measurements from a Vancouver epidemiologic study. Measured ultrafine PM and PM(2.5) concentration levels along the residential urban street canyon and at the test house facade in Sunset Park were demonstrated to be comparable to traffic levels at an arterial road and slightly higher than those in a residential area of Los Angeles. Indoor ultrafine PM levels were roughly 3-10 times lower than outdoor levels, depending on the monitor location. CO, NO(2), and SO(2) levels were shown to be similar to values that produced increased risk of chronic obstructive pulmonary disease hospitalizations in the Vancouver studies. PMID- 20024010 TI - Characterization of traffic-related PM concentration distribution and fluctuation patterns in near-highway urban residential street canyons. AB - Analyses of outdoor traffic-related particulate matter (PM) concentration distribution and fluctuation patterns in urban street canyons within a microscale distance of less than 500 m from a highway source are presented as part of the results from the Brooklyn Traffic Real-Time Ambient Pollutant Penetration and Environmental Dispersion (B-TRAPPED) study. Various patterns of spatial and temporal changes in the street canyon PM concentrations were investigated using time-series data of real-time PM concentrations measured during multiple monitoring periods. Concurrent time-series data of local street canyon wind conditions and wind data from the John F. Kennedy (JFK) International Airport National Weather Service (NWS) were used to characterize the effects of various wind conditions on the behavior of street canyon PM concentrations.Our results suggest that wind direction may strongly influence time-averaged mean PM concentration distribution patterns in near-highway urban street canyons. The rooftop-level wind speeds were found to be strongly correlated with the PM concentration fluctuation intensities in the middle sections of the street blocks. The ambient turbulence generated by shifting local wind directions (angles) showed a good correlation with the PM concentration fluctuation intensities along the entire distance of the first and second street blocks only when the wind angle standard deviations were larger than 30 degrees. Within canyon turbulent shearing, caused by fluctuating local street canyon wind speeds, showed no correlation with PM concentration fluctuation intensities. The time averaged mean PM concentration distribution along the longitudinal distances of the street blocks when wind direction was mostly constantly parallel to the street was found to be similar to the distribution pattern for the entire monitoring period when wind direction fluctuated wildly. Finally, we showed that two different PM concentration metrics-time-averaged mean concentration and number of concentration peaks above a certain threshold level-can possibly lead to different assessments of spatial concentration distribution patterns. PMID- 20024011 TI - Establishing a link between vehicular PM sources and PM measurements in urban street canyons. AB - The Brooklyn Traffic Real-Time Ambient Pollutant Penetration and Environmental Dispersion (B-TRAPPED) study, conducted in Brooklyn, NY, USA, in 2005, was designed with multiple goals in mind, two of which were contaminant source characterization and street canyon transport and dispersion monitoring. In the portion of the study described here, synchronized wind velocity and azimuth as well as particulate matter (PM) concentrations at multiple locations along 33rd Street were used to determine the feasibility of using traffic emissions in a complex urban topography as a sole tracer for studying urban contaminant transport. We demonstrate in this paper that it is possible to link downwind concentrations of contaminants in an urban street canyon to the vehicular traffic cycle using Eigen-frequency analysis. In addition, multivariable circular histograms are used to establish directional frequency maxima for wind velocity and contaminant concentration. PMID- 20024012 TI - Time-series analysis to study the impact of an intersection on dispersion along a street canyon. AB - This paper presents data analysis from the Brooklyn Traffic Real-Time Ambient Pollutant Penetration and Environmental Dispersion (B-TRAPPED) study to assess the transport of ultrafine particulate matter (PM) across urban intersections. Experiments were performed in a street canyon perpendicular to a highway in Brooklyn, NY, USA. Real-time ultrafine PM samplers were positioned on either side of an intersection at multiple locations along a street to collect time-series number concentration data. Meteorology equipment was positioned within the street canyon and at an upstream background site to measure wind speed and direction. Time-series analysis was performed on the PM data to compute a transport velocity along the direction of the street for the cases where background winds were parallel and perpendicular to the street. The data were analyzed for sampler pairs located (1) on opposite sides of the intersection and (2) on the same block. The time-series analysis demonstrated along-street transport, including across the intersection when background winds were parallel to the street canyon and there was minimal transport and no communication across the intersection when background winds were perpendicular to the street canyon. Low but significant values of the cross-correlation function (CCF) underscore the turbulent nature of plume transport along the street canyon. The low correlations suggest that flow switching around corners or traffic-induced turbulence at the intersection may have aided dilution of the PM plume from the highway. This observation supports similar findings in the literature. Furthermore, the time-series analysis methodology applied in this study is introduced as a technique for studying spatiotemporal variation in the urban microscale environment. PMID- 20024013 TI - The effect of a tall tower on flow and dispersion through a model urban neighborhood: part 1. Flow characteristics. AB - Wind tunnel experiments were performed to examine the effect of a tall tower on the flow around an otherwise uniform array of buildings. Additionally, preliminary CFD simulations were run to visualize the flow with more resolution. The model used in both the wind tunnel and CFD studies was designed to simulate an area of Brooklyn, NY, USA, where blocks of residential row houses form a neighborhood bordering a major urban highway. This area was the site of a field study that, along with the work reported here, had the goal of improving the understanding of airflow and dispersion patterns within urban microenvironments. Results reveal that a tall tower has a dramatic effect on the flow in the street canyons in the neighboring blocks, enhancing the exchange between the street canyon flow and the freestream flow aloft. In particular, vertical motion down the windward side and up the leeward side of the tower resulted in strong flows in the lateral street canyons and increased winds in the street canyons in the immediate vicinity of the tower. These phenomena were visible in both the wind tunnel and CFD results, although some minor differences in the flow fields were noted. PMID- 20024014 TI - The effect of a tall tower on flow and dispersion through a model urban neighborhood: part 2. Pollutant dispersion. AB - This article is the second in a two-paper series presenting results from wind tunnel and computational fluid dynamics (CFD) simulations of flow and dispersion in an idealized model urban neighborhood. Pollutant dispersion results are presented and discussed for a model neighborhood that was characterized by regular city blocks of three-story row houses with a single 12-story tower located at the downwind edge of one of these blocks. The tower had three significant effects on pollutant dispersion in the surrounding street canyons: drawing the plume laterally towards the tower, greatly enhancing the vertical dispersion of the plume in the wake of the tower, and significantly decreasing the residence time of pollutants in the wake of the tower. In the wind tunnel, tracer gas released in the avenue lee of the tower, but several blocks away laterally, was pulled towards the tower and lifted in the wake of the tower. The same lateral movement of the pollutant was seen in the next avenue, which was approximately 2.5 tower heights downwind of the tower. The tower also served to ventilate the street canyon directly in its wake more rapidly than the surrounding areas. This was evidenced by CFD simulations of concentration decay where the residence time of pollutants lee of the 12-story tower was found to be less than half the residence time behind a neighboring three-story building. This same phenomenon of rapid vertical dispersion lee of a tower among an array of smaller buildings was also demonstrated in a separate set of wind tunnel experiments using an array of cubical blocks. A similar decrease in the residence time was observed when the height of one block was increased. PMID- 20024015 TI - Building characterization and aerosol infiltration into a naturally ventilated three-story apartment building. AB - Understanding infiltration of outdoor pollutants was an integral part of the Brooklyn Traffic Real-Time Ambient Pollutant Penetration and Environmental Dispersion (B-TRAPPED) study. For this reason, the structural and air exchange properties of the three-story row house in Brooklyn, NY, USA, that was used in the B-TRAPPED experiments were fully characterized. Factors investigated included representativeness of the construction and impact of building design features on the natural ventilation and infiltration of outdoor aerosol. Both blower door and perfluorocarbon tracer (PFT) air exchange rate (AER) experiments showed that the ventilation rates of the building were quite typical of similar structures in the New York City (NYC) metropolitan area. Indoor/outdoor (I/O) aerosol count ratios by particle size were comparable to a similar vintage naturally ventilated building in Boston, MA, USA. I/O ratio analyses were consistent with literature findings and showed I/O ratios ranging from 0.310 to 0.601, varying across particle sizes (from 0.3 to 5.0 [corrected] mum) and between first and second floor apartments. An effort to apply the rebound method of Thatcher et al. (Aerosol Sci. Technol., 2003, 37, 847-864) in determining aerosol infiltration rates proved unsuccessful due to unexpectedly long (>60 min) equilibration times after the filtration period. Uninsulated interior wall renovations in the study house created a cavity that resulted in a large intermediate dead volume (for infiltration) that apparently could not be accommodated by a simple infiltration model. Simple two-compartment models evidently have finite application limitations for even modestly complex settings. PMID- 20024016 TI - Parameterization of meteorological variables in the process of infiltration of outdoor ultrafine particles into a residential building. AB - As part of the Brooklyn Traffic Real-Time Ambient Pollutant Penetration and Environmental Dispersion (B-TRAPPED) study, a field investigation was conducted of the mechanisms involved in infiltration of outdoor particles (0.02 microm to 1 microm) into a near-highway urban residential building. Using continuous real time total number concentration time-series data measured simultaneously at multiple outdoor and indoor locations, the infiltration time was estimated for various indoor sites by using the cross-correlation analysis method. The effects of meteorological variables on infiltration times were also characterized at given locations. In particular, infiltration time was examined as a function of outdoor mean rooftop wind speed. Outdoor and indoor temperature and relative humidity were also investigated in relation to infiltration times. Our results showed that outdoor wind speed was the dominant meteorological parameter affecting the infiltration rate, but no correlation was found with temperature. The outdoor-indoor PM concentration ratio was found to be significantly different for the first and second floors of the test building. Finally, we determined the effectiveness of a protective shelter-in-place area inside the building by calculating infiltration times into the area. We found that the shelter-in-place area offered some protection against the infiltrating particles because of increased attenuation of the outdoor concentration. PMID- 20024017 TI - Analysis of indoor air pollution trends and characterization of infiltration delay time using a cross-correlation method. AB - High-density housing in close proximity to freeways in conjunction with high concentrations of traffic emissions may contribute to significant degradation of indoor air quality. Densely populated areas may also be targeted for intentional releases of biological or chemical agents because an urban release could result in higher morbidity and mortality from the attack. Since people tend to spend the majority of their time indoors, it is paramount to explore the relationships between outdoor and indoor air quality and, specifically, the time scales that characterize transport of airborne contaminants from outdoors to indoors. In the Brooklyn Traffic Real-Time Ambient Pollutant Penetration and Environmental Dispersion (B-TRAPPED) study, a three-story row house with a flat face and roof and multiple rooms was used to investigate outdoor-to-indoor contaminant time scales. The building was located in the Sunset Park neighborhood of Brooklyn, NY, USA, in the vicinity of a major expressway and a heavily trafficked arterial road. It was found that the building shell has a profound impact on the indoor concentrations. A strong hourly periodicity (see Eisner et al., this issue, DOI: 10.1039/b907132f) in concentration outside the building during the morning "rush hour" was used as evidence to suggest that indoor contaminants originated from outdoor air penetration. Although the indoor concentrations followed a similar pattern, indoor concentrations were found to be more persistent than outdoor concentrations. Stronger persistency is used here to describe the tendency of the indoor concentration to continue to rise even if the outdoor concentration has started to drop, or vice versa. This may be an important factor in assessing negative health risks to inhabitants or first responders. A cross-correlation technique was employed to study the correlation between outdoor and indoor time series. In the high-density housing residential building used in the study, it was found that a long lag time exists (11 min) before indoor and outdoor concentrations reach maximal correlation. PMID- 20024018 TI - Fate of organohalogens in US wastewater treatment plants and estimated chemical releases to soils nationwide from biosolids recycling. AB - This study examined the occurrence in wastewater of 11 aromatic biocides, pesticides and degradates, and their fate during passage through US treatment plants, as well as the chemical mass contained in sewage sludge (biosolids) destined for land application. Analyte concentrations in wastewater influent, effluent and sludge from 25 facilities in 18 US states were determined by liquid chromatography electrospray (tandem) mass spectrometry. Dichlorocarbanilide, fipronil, triclocarban, and triclosan were found consistently in all sample types. Dichlorophene, hexachlorophene, and tetrachlorocarbanilide were detected infrequently only, and concentrations of the phenyl urea pesticides diflubenzuron, hexaflumuron, and linuron were below the limit of detection in all matrixes. Median concentrations (+/-95% confidence interval) of quantifiable compounds in influent ranged from 4.2 +/- 0.8 microg L(-1) for triclocarban to 0.03 +/- 0.01 microg L(-1) for fipronil. Median concentrations in effluent were highest for triclocarban and triclosan (0.23 +/- 0.08 and 0.07 +/- 0.04 microg L( 1), respectively). Median aqueous-phase removal efficiencies (+/-95% CI) of activated sludge treatment plants decreased in the order of: triclosan (96 +/- 2%) > triclocarban (87 +/- 7%) > dichlorocarbanilide (55 +/- 20%) > fipronil (18 +/- 22%). Median concentrations of organohalogens were typically higher in anaerobically than in aerobically digested sludges, and peaked at 27 600 +/- 9600 and 15 800 +/- 8200 microg kg(-1) for triclocarban and triclosan, respectively. Mass balances obtained for three primary pesticides in six activated sludge treatment plants employing anaerobic digestion suggested a decreasing overall persistence from fipronil (97 +/- 70%) to triclocarban (87 +/- 29%) to triclosan (28 +/- 30%). Nationwide release of the investigated organohalogens to agricultural land via municipal sludge recycling and into surface waters is estimated to total 258 000 +/- 110 00 kg year(-1) (mean +/- 95% confidence interval), with most of this mass derived from antimicrobial consumer products of daily use. This study addresses some of the data gaps identified by the National Research Council in its 2002 study on standards and practices of biosolids application on land. PMID- 20024019 TI - The determination of nitrogen dioxide in ambient air with free hanging filters as passive samplers, and a new calibration method using fritted bubblers. AB - This paper describes two new methods for the determination of NO(2) in the ambient air. The first method consists of free hanging filters with a diameter of 2.5 cm as passive samplers. The filters are impregnated with triethanolamine to bind NO(2). With standard colorimetrical analysis, the amount of NO(2) on the filters is determined. The second method is performed with fritted bubblers filled with Saltzman reagent, where, with a special procedure the absorption efficiencies of the bubblers are determined using ambient air, without the use of standard gases and electronic analytical instruments. The results of the bubblers are used to calibrate the free hanging filters. The two methods were applied simultaneously in the city of Yogyakarta, Indonesia. The methods are inexpensive and very well suited for use in low-budget situations. A characteristic of the free filter is the Sampling Volume, SV. This is the ratio of the amount of NO(2) on the filter and the ambient concentration. With the filter used in this study, the amount of triethanolamine and exposure time, the SV is 0.0166 m(3). The sampling rate (SR) of the filter, 4.6 cm(3)/s, is high. Hourly averaged measurements are performed for 15 hours per day in four busy streets. The measured amounts of NO(2) on the filters varied between 0.57 and 2.02 microg NO(2), at ambient air concentrations of 32 to 141 microg/m(3) NO(2). During the experiments the wind velocity was between 0.2 and 2.0 m/s, the relative humidity between 24 and 83 % and the temperature between 295 K and 311 K. These variations in weather conditions have no influence on the uptake of NO(2). PMID- 20024020 TI - Chemotrapping-atomic fluorescence spectrometric method as a field method for volatile arsenic in natural gas. AB - Volatile arsenic compounds in natural gas, existing in the form of trimethylarsine (TMAs), have been determined using gas cryo-trapping gas chromatography coupled to inductively coupled plasma-mass spectrometry (CT-GC-ICP MS). The results from a number of different gas wells revealed a huge concentration spread ranging from below the detection limit of 0.2 up to 1800 microg/m(3) TMAs (as As) in the gas. Due to the toxicity and corrosive nature of these arsines, they need near real time monitoring via a method that can easily be implemented on site, i.e. during gas exploitation. Here, we introduce a novel method which utilises silver nitrate impregnated silica gel tubes for quantitative chemotrapping of trimethylarsine (TMAs) from a natural gas matrix. Subsequent elution with hot nitric acid followed by online photo-oxidation hydride generation atomic fluorescence spectrometry (HG-AFS) is used for the determination of TMAs gas standards in nitrogen and natural gas samples, respectively. The chemotrapping method was validated using CT-GC-ICP-MS as a reference method. The recovery of arsenic from nitrogen or natural gas matrix ranged from 85 to 113% for a range of 20 to 2000 ng As. Trapping efficiency was >98%, from the methods LOD of 20 ng to 4.8 microg (absolute amount As) with sample sizes of 0.02 and 2 L gas. Method performance was established by comparing the results obtained for eight natural gas samples containing between 1 and 140 microg As/m(3) with those achieved by the reference method (CT-GC-ICP-MS). PMID- 20024021 TI - Vegetation greening in the Canadian Arctic related to decadal warming. AB - This study is presented within the context that climate warming and sea-ice decline has been occurring throughout much of the Arctic over the past several decades, and that terrestrial ecosystems at high latitudes are sensitive to the resultant alterations in surface temperatures. Results are from analyzing interannual satellite records of vegetation greenness across a bioclimate gradient of the Canadian Arctic over the period of 1982-2006. Here, we combine multi-scale sub-pixel analysis and remote sensing time-series analysis to investigate recent decadal changes in vegetation greenness along spatial gradients of summer temperature and vegetation. Linear autoregression temporal analysis of vegetation greenness was performed with relatively "pure" vegetation pixels of Advanced Very High Resolution Radiometer (AVHRR) data, spanning Low Arctic, High Arctic and polar desert ecosystems. Vegetation greenness generally increased over tundra ecosystems in the past two decades. Peak annual greenness increased 0.49-0.79%/yr over the High Arctic where prostrate dwarf shrubs, forbs, mosses and lichens dominate and 0.46-0.67%/yr over the Low Arctic where erect dwarf shrubs and graminoids dominate. However, magnitudes of vegetation greenness differ with length of time series and periods considered, indicating a nonlinear response of terrestrial ecosystems to climate change. The decadal increases of greenness reflect increasing vegetation production during the peak of the growing season, and were likely driven by the recent warming. PMID- 20024024 TI - PCR amplification and genetic analysis in a microwell cell culturing chip. AB - We have previously described a microwell chip designed for high throughput, long term single-cell culturing and clonal analysis in individual wells providing a controlled way of studying high numbers of individual adherent or non-adherent cells. Here we present a method for the genetic analysis of cells cultured on chip by PCR and minisequencing, demonstrated using two human adherent cell lines: one wild type and one with a single-base mutation in the p53 gene. Five wild type or mutated cells were seeded per well (in a defined set of wells, each holding 500 nL of culture medium) in a 672-microwell chip. The cell chip was incubated overnight, or cultured for up to five days, depending on the desired colony size, after which the cells were lysed and subjected to PCR directly in the wells. PCR products were detected, in the wells, using a biotinylated primer and a fluorescently labelled primer, allowing the products to be captured on streptavidin-coated magnetic beads and detected by a fluorescence microscope. In addition, to enable genetic analysis by minisequencing, the double-stranded PCR products were denatured and the immobilized strands were kept in the wells by applying a magnetic field from the bottom of the wells while the wells were washed, a minisequencing reaction mixture was added, and after incubation in appropriate conditions the expected genotypes were detected in the investigated microwells, simultaneously, by an array scanner. We anticipate that the technique could be used in mutation frequency screening, providing the ability to correlate cells' proliferative heterogeneity to their genetic heterogeneity, in hundreds of samples simultaneously. The presented method of single-cell culture and DNA amplification thus offers a potentially powerful alternative to single-cell PCR, with advantageous robustness and sensitivity. PMID- 20024025 TI - Field-effect based attomole titrations in nanoconfinement. AB - This paper describes a novel capacitive method to change the pH in micro- and nanofluidic channels. A device with two metal gate electrodes outside an insulating channel wall is used for this purpose. The device is operated at high ionic strength with thin double layers. We demonstrate that gate potentials applied between the electrodes cause a release or uptake of protons from the silicon nitride surface groups, resulting in a pH shift in the channel and a titration of solution compounds present. Due to the high quality silicon nitride insulating layer, the effect is purely capacitive and electrolysis can be neglected. Fluorescein was employed as a fluorescent pH indicator to quantify the induced pH changes, and a maximum change of 1.6 pH units was calculated. A linear relationship was found between applied potential and fluorescein intensity change, indicating a linear relation between actuated proton amount and applied voltage. Since this pH actuation method avoids redox reactions and can be operated at physiological ionic strength, it can be very useful as a "soft" way to change the pH in very small volumes e.g. in bioassays or cell-based research. The sensitivity of the optical detection method poses the only limit to the detectable amount of substance and the observed volume. In a preliminary measurement we show one possible application, namely titration of 100 attomol of TRIS in a 7 pL detection volume. It is important to stress that this pH actuation principle fundamentally differs from the pH changes occurring in ionic transistors which are due to counterion enrichment and coion exclusion, because it does not rely on double-layer overlap. As a result it can be operated at high ionic strength and in channels of up to at least 1 microm height. PMID- 20024026 TI - Room-temperature intermediate layer bonding for microfluidic devices. AB - In this work a novel room-temperature bonding technique based on chemically activated Fluorinated Ethylene Propylene (FEP) sheet as an intermediate between chemically activated substrates is presented. Surfaces of silicon and glass substrates are chemically modified with APTES bearing amine terminal groups, while FEP sheet surfaces are treated to form carboxyl groups and subsequently activated by means of EDC-NHS chemistry. The activation procedures of silicon, glass and FEP sheet are characterized by contact angle measurements and XPS. Robust bonds are created at room-temperature by simply pressing two amine terminated substrates together with activated FEP sheet in between. Average tensile strengths of 5.9 MPa and 5.2 MPa are achieved for silicon-silicon and glass-glass bonds, respectively, and the average fluidic pressure that can be operated is 10.2 bar. Moreover, it is demonstrated that FEP-bonded microfluidic chips can handle mild organic solvents at elevated pressures without leakage problems. This versatile room-temperature intermediate layer bonding technique has a high potential for bonding, packaging, and assembly of various (bio-) chemical microfluidic systems and MEMS devices. PMID- 20024027 TI - "Print-n-Shrink" technology for the rapid production of microfluidic chips and protein microarrays. AB - An innovative method for the production of microfluidic chips integrating protein spots is described. The technology, called "Print-n-Shrink", is based on the screen-printing of a microfluidic design (using a dielectric ink) onto Polyshrink polystyrene sheets. The initial print which has a minimum size of 15 microm (height) x 230 microm (width) is thermally treated (30 seconds, 163 degrees C) to shrink and generate features of 85 microm (height) x 100 microm (width). Concomitantly, proteins such as monoclonal antibodies or cellular adhesion proteins are spotted onto the Polyshrink sheets and shrunk together with the microfluidic design, creating a complete biochip integrating both complex microfluidic designs and protein spots for bioanalytical applications. PMID- 20024028 TI - A blood sampling microsystem for pharmacokinetic applications: design, fabrication, and initial results. AB - This paper describes a microsystem for automated blood sampling from laboratory mice used in pharmacokinetic studies. Intended to be mounted as a "backpack" on a mouse, it uses a microneedle, reservoir, and an actuator to instantaneously prick the animal for a time-point sample, eliminating the need for a tethered catheter with large dead volume. The blood is collected by capillary effect through a 31 33 gauge microneedle (250-210 microm OD) into a approximately 1 microL micromachined steel reservoir. The voice coil actuator provides a peak force of approximately 300 mN, which amply exceeds the measured piercing force of mouse skin (i.e., 60-85 mN for a 31-gauge needle with 12 degrees bevel). The sampling system was tested in vitro using a mock vessel with adjustable pressure; the reservoir was filled in <0.15 s by a combination of the capillary effect and blood pressure. The system may also be used to sample interstitial fluid, but the absence of blood pressure makes it necessary to enhance the capillary effect of the needle. This is accomplished by either electropolishing the inner surface to make it more hydrophilic or using a polymer wire insert to increase the surface area. The steel surface of the reservoir is also coated with silicon oxynitride by plasma-enhanced chemical vapor deposition to improve its hydrophilicity. Blood from fresh bovine tissue was collected into the reservoir to simulate interstitial fluid sampling. In vivo tests on live, anesthetized mice resulted in successful collection of blood into the reservoir. The possible integration of the device in microanalytical systems and the device scalability for multisampling are discussed. PMID- 20024029 TI - Rapid integrated biosensor for multiplexed immunoassays based on actuated magnetic nanoparticles. AB - The realization of biomolecular detection assays for diagnostic purposes is technologically very challenging because such tests demand full integration for ease of use and need to deliver a high analytical performance with cost-effective use of materials. In this article an optomagnetic immunoassay technology is described based on nanoparticles that are magnetically actuated and optically detected in a stationary sample fluid. The dynamic control of nanoparticles by magnetic fields impacts the key immunoassay process steps, giving unprecedented speed, assay control and seamless integration of the total test. The optical detection yields sensitive and multiplexed assays in a low-cost disposable cartridge. We demonstrate that the optomagnetic technology enables high sensitivity one-step assays in blood serum/plasma and whole saliva. Drugs of abuse are detected at sub-nanogram per millilitre levels in a total assay time of 1 min, and the cardiac marker troponin I is detected at sub-picomole per litre concentrations in a few minutes. The optomagnetic technology is fundamentally suited for high-performance integrated testing and is expected to open a new paradigm in biosensing. PMID- 20024030 TI - High-pressure on-chip mechanical valves for thermoplastic microfluidic devices. AB - A facile method enabling the integration of elastomeric valves into rigid thermoplastic microfluidic chips is described. The valves employ discrete plugs of elastomeric polydimethylsiloxane (PDMS) integrated into the thermoplastic substrate and actuated using a threaded stainless steel needle. The fabrication process takes advantage of poly(ethylene glycol) (PEG) as a sacrificial molding material to isolate the PDMS regions from the thermoplastic flow channels, while yielding smooth contact surfaces with the PDMS valve seats. The valves introduce minimal dead volumes, and provide a simple mechanical means to achieve reproducible proportional valving within thermoplastic microfluidic systems. Burst pressure tests reveal that the valves can withstand pressures above 12 MPa over repeated open/close cycles without leakage, and above 24 MPa during a single use, making the technology well suited for applications such as high performance liquid chromatography. Proportional valve operation is demonstrated using a multi valve chemical gradient generator fabricated in cyclic olefin polymer. PMID- 20024031 TI - Correlating short-term Ca(2+) responses with long-term protein expression after activation of single T cells. AB - In order to elucidate the dynamics of cellular processes that are induced in context with intercellular communication, defined events along the signal transduction cascade and subsequent activation steps have to be analyzed on the level of individual cells and correlated with each other. Here we present an approach that allows the initiation of cell-cell or cell-particle interactions and the analysis of cellular reactions within various regimes while the identity of each individual cell is preserved. It utilizes dielectrophoresis (DEP) and microfluidics in a lab-on-chip system. With high spatial and temporal precision we contacted single T cells with functionalized microbeads and monitored their immediate cytosolic Ca(2+) response. After this, the cells were released from the chip system and cultivated further. Expression of the activation marker molecule CD69 was analyzed the next day and correlated with the previously recorded Ca(2+) signal for each individual cell. We found a significant difference in the patterns of Ca(2+) traces between activated and non-activated cells, which shows that Ca(2+) signals in T cells can provide early information about a later reaction of the cell. Although T cells are non-excitable cells, we also observed irregular Ca(2+) transients upon exposure to the DEP field only. These Ca(2+) signals depended on exposure time, electric field strength and field frequency. By minimizing their occurrence rate, we could identify experimental conditions that caused the least interference with the physiology of the cell. PMID- 20024032 TI - 2D-PCR: a method of mapping DNA in tissue sections. AB - A novel approach was developed for mapping the location of target DNA in tissue sections. The method combines a high-density, multi-well plate with an innovative single-tube procedure to directly extract, amplify, and detect the DNA in parallel while maintaining the two-dimensional (2D) architecture of the tissue. A 2D map of the gene glyceraldehyde 3-phosphate dehydrogenase (GAPDH) was created from a tissue section and shown to correlate with the spatial area of the sample. It is anticipated that this approach may be easily adapted to assess the status of multiple genes within tissue sections, yielding a molecular map that directly correlates with the histology of the sample. This will provide investigators with a new tool to interrogate the molecular heterogeneity of tissue specimens. PMID- 20024033 TI - Microfluorimeter with disposable polymer chip for detection of coeliac disease toxic gliadin. AB - Coeliac disease is an inflammatory disease of the upper small intestine and results from gluten ingestion in genetically susceptible individuals, and is the only life-long nutrient-induced enteropathy. The only treatment is a strict gluten-free diet and the longer the individual fails to adhere to this diet, the greater the chance of developing malnutrition and other complications. The existence of reliable gluten free food is crucial to the well-being of the population. Here we report on a microfluorimeter device for the in situ detection of gliadin in foodstuffs, which could be used for a rapid control of raw materials in food processing, as well as for process control of gliadin contamination. The microfluorimeter is based on a reflector that is used inside a microfluidic chip, exploiting various strategically placed reflective or totally metallised mirrors for efficient collection of the fluorescent light emitted in a large solid angle. The chip is capable of executing five assays in parallel and has been demonstrated to possess detection sensitivity applicable to fluoroimmunoassays. Various immunoassay formats exploiting fluorescence detection, using enzyme/fluorophore labels were developed and compared in terms of sensitivity, ease of assay, assay time and compatibility with buffer used to extract gliadin from raw and cooked foodstuffs, with the best performance observed with an indirect competition assay using a fluorophore-labelled anti mouse antibody. This assay was exploited within the microfluorimeter device, and a very low detection limit of 4.1 ng/mL was obtained. The system was observed to be highly reproducible, with an RSD of 5.9%, for a concentration of 50 ng/mL of gliadin applied to each of the five channels of the microfluorimeter. Biofunctionalised disposable strips incorporated into the microfluorimeter were subjected to accelerated Arrhenius thermal stability studies and it was demonstrated that strips pre-coated with gliadin could be stored for approximately 2 years at 4 degrees C, with no discernable loss in sensitivity or detectability of the assay. Finally, the microfluorimeter was applied to the analysis of commercial gluten-free food samples, and an excellent correlation with routine ELISA measurements was obtained. The developed microfluorimeter should find widespread application for on-site execution of fluoroimmunoassays. PMID- 20024034 TI - Rapid protein depletion from complex samples using a bead-based microfluidic device for the point of care. AB - Translation of sample preparation methods to point-of-care formats has remained a challenge. We present a plastic laminate microfluidic device for protein depletion from human plasma using ligand immobilized porous beads stored dry within a novel, pneumatically-driven mixer. The card design accelerated the protein depletion process from hours to minutes. Using immunoglobulin G as a model protein, we have successfully shown protein removal efficiency from spiked buffer between 70-80% and from diluted human plasma samples between 66-77%. Low non-specific binding of our downstream target ligand, immunoglobulin M, was observed with the spiked buffer and diluted human plasma samples. For future device optimization, the physical limitations to rapid protein removal on card were also explored. Bench-top experiments with improved mixing efficiency and a lower sample dilution factor achieved 99% IgG removal using the same amount of mixing time. This design can easily be adapted for depletion of other high abundance or interfering proteins by inclusion of other ligand immobilized beads. PMID- 20024035 TI - Detection of viruses with molecularly imprinted polymers integrated on a microfluidic biochip using contact-less dielectric microsensors. AB - Rapid detection of viral contamination remains a pressing issue in various fields related to human health including clinical diagnostics, the monitoring of food borne pathogens, the detection of biological warfare agents as well as in viral clearance studies for biopharmaceutical products. The majority of currently available assays for virus detection are expensive, time-consuming, and labor intensive. In the present work we report the creation of a novel micro total analysis system (microTAS) capable of continuously monitoring viral contamination with high sensitivity and selectivity. The specific interaction between shape and surface chemistry between molecular imprinted polymer (MIP) and virus resulted in the elimination of non-specific interaction in the present sensor configuration. The additional integration of the blank (non-imprinted) polymer further allowed for the identification of non-specific adsorption events. The novel combination of microfluidics containing integrated native polymer and MIP with contact-less dielectric microsensors is evaluated using the Tobacco Mosaic Virus (TMV) and the Human Rhinovirus serotype 2 (HRV2). Results show that viral binding and dissociation events can be readily detected using contact-less bioimpedance spectroscopy optimized for specific frequencies. In the present study optimum sensor performance was achieved at 203 kHz within the applied frequency range of 5-500 kHz. Complete removal of the virus from the MIP and device reusability is successfully demonstrated following a 50-fold increase in fluid velocity. Evaluation of the microfluidic biochip revealed that microchip technology is ideally suited to detect a broader range of viral contaminations with high sensitivity by selectively adjusting microfluidic conditions, sensor geometries and choice of MIP polymeric material. PMID- 20024037 TI - Sequential processing from cell lysis to protein assay on a chip enabling the optimization of an F(1)-ATPase single molecule assay condition. AB - We developed an integrated protein assay device, "Single Molecule MicroTAS (SMM)," which enables cell lysis, protein extraction, purification, and activity assay. The assay was achieved at the single-molecule scale for a genetically engineered protein, F(1)-ATPase, which is the smallest known rotary motor. A cell lysis condition, with a wide range of applied voltages (50-250 V) and other optimized values (pulse width: 50 micros; duty: 0.01%; electrode gap: 25 microm; total flow rate: 5 microL min(-1)) provided a high enough protein concentration for the assay. Successively, the protein was extracted and purified by specific binding in a microfluidic channel. During the assay process, the diffusion effect of lysate between a two-phase laminar flow contributes to optimizing the single molecule assay condition, because the concentration of the original lysate from the E. coli solution is too high to assay. To achieve the most efficient assay condition, the protein diffusion effect on the assay was experimentally and numerically evaluated. The results reveal that, in our experimental conditions, concentrations of F(1) and other contaminated effluents are optimized for the F(1) rotational assay at a channel position. The adenosine triphosphate (ATP) driven rotation speed measured in the SMM was compatible with that obtained by conventional purification and assay. Such a sequential process from cell lysis to assay proves that the SMM is an example of a sample-in-answer-out system for F(1) protein evaluation. PMID- 20024038 TI - Multifunctional plasmonic sensors on low-cost subwavelength metallic nanoholes arrays. AB - Localized surface plasmon resonance (LSPR) sensing is combined with surface enhanced Raman scattering (SERS) detection on periodic arrays of subwavelength metallic nanoholes for the first time. LSPR sensors provide detection of molecular adsorption in both transmission and reflectivity modes with sensitivity greater that 300 nm/RIU and a spectral shift of 16 nm in the case of binding a monolayer of p-aminothiophenol (p-ATP) molecules. Subsequent SERS analysis enables identification of the adsorbed p-ATP molecule, its structure and orientation on the metal surface. This synergistic LSPR-SERS approach on low-cost metallic films perforated with periodic arrays of subwavelength nanoholes opens a route for molecular dual-modal detection to be integrated in lab-on-chip systems to increase the reliability of biological detection. PMID- 20024036 TI - Highly parallel introduction of nucleic acids into mammalian cells grown in microwell arrays. AB - High-throughput cell-based screens of genome-size collections of cDNAs and siRNAs have become a powerful tool to annotate the mammalian genome, enabling the discovery of novel genes associated with normal cellular processes and pathogenic states, and the unravelling of genetic networks and signaling pathways in a systems biology approach. However, the capital expenses and the cost of reagents necessary to perform such large screens have limited application of this technology. Efforts to miniaturize the screening process have centered on the development of cellular microarrays created on microscope slides that use chemical means to introduce exogenous genetic material into mammalian cells. While this work has demonstrated the feasibility of screening in very small formats, the use of chemical transfection reagents (effective only in a subset of cell lines and not on primary cells) and the lack of defined borders between cells grown in adjacent microspots containing different genetic material (to prevent cell migration and to aid spot location recognition during imaging and phenotype deconvolution) have hampered the spread of this screening technology. Here, we describe proof-of-principles experiments to circumvent these drawbacks. We have created microwell arrays on an electroporation-ready transparent substrate and established procedures to achieve highly efficient parallel introduction of exogenous molecules into human cell lines and primary mouse macrophages. The microwells confine cells and offer multiple advantages during imaging and phenotype analysis. We have also developed a simple method to load this 484-microwell array with libraries of nucleic acids using a standard microarrayer. These advances can be elaborated upon to form the basis of a miniaturized high-throughput functional genomics screening platform to carry out genome-size screens in a variety of mammalian cells that may eventually become a mainstream tool for life science research. PMID- 20024039 TI - Rapid multicomponent optical protein patterning. AB - Cells sense spatial distributions of molecules which trigger signal transduction pathways that induce the cell to migrate or extend by remodelling the cytoskeleton. However, the influence of local and small variations of extracellular protein concentration on chemotaxis is not fully understood, due in part to the lack of simple and precise methods to pattern proteins in vitro. We recently developed a new technology to fabricate such patterns which relies on photobleaching fluorophores to adsorb proteins on a cell culture substrate: laser assisted protein adsorption by photobleaching (LAPAP). Here we report several key improvements to LAPAP: we created arbitrary patterns made of several different proteins simultaneously, we reduced the fabrication time more than one order of magnitude and we used secondary antibodies to significantly enlarge the spectrum of proteins that can be employed. As a result, multicomponent protein gradients can be produced using reagents that are typically available in life science research laboratories on a standard inverted microscope equipped with a camera port. PMID- 20024040 TI - Micro contactor based on isotachophoretic sample transport. AB - It is demonstrated how isotachophoresis (ITP) in a microfluidic device may be utilized to bring two small sample volumes into contact in a well-controlled manner. The ITP contactor serves a similar purpose as micromixers that are designed to mix two species rapidly in a microfluidic channel. In contrast to many micromixers, the ITP contactor does not require complex channel architectures and allows a sample processing in the spirit of "digital microfluidics", i.e. the samples always remain in a compact volume. It is shown that the ITP zone transport through microchannels proceeds in a reproducible and predictable manner, and that the sample trajectories follow simple relationships obtained from Ohm's law. Firstly, the micro contactor can be used to synchronize two ITP zones having reached a channel at different points in time. Secondly, fulfilling its actual purpose it is capable of bringing two samples in molecular contact via an interpenetration of ITP zones. It is demonstrated that the contacting time is proportional to the ITP zone extension. This opens up the possibility of using that type of device as a special type of micromixer with "mixing times" significantly below one second and an option to regulate the duration of contact through specific parameters such as the sample volume. Finally, it is shown how the micro contactor can be utilized to conduct a hybridization reaction between two ITP zones containing complementary DNA strands. PMID- 20024041 TI - An automated, pre-programmed, multiplexed, hydraulic microvalve. AB - An automated, pre-programmed, multiplexed hydraulic valve actuator is described. The valve is membrane-based and normally open. In contrast to the membrane-based pneumatic valve, the hydraulic valve uses hydraulic liquid to exert the control pressure. The line pressure is controlled with a roller moving over a prefabricated topology. Multiple rollers, each traversing its own track, are assembled into a single carriage, which can be actuated either manually or with a single computer-controlled motor. A valve manifold and roller actuators are designed, fabricated, and tested to demonstrate three-way valve actuation in a pre-determined sequence. The performance of the valve is evaluated and the utility of the valve in the operation of a micro thermal cycler was demonstrated. Hydraulic controllers of the type described here can be operated either manually or under computer control and provide an inexpensive means of controlling flow in lab-on-a-chip devices. PMID- 20024042 TI - Centrifugo-pneumatic valve for metering of highly wetting liquids on centrifugal microfluidic platforms. AB - We designed and experimentally validated a new type of passive valve for centrifugal microfluidic platforms. A liquid column entering an unvented receiving chamber is stopped by the counter-pressure of compressed air. This valve opens under defined conditions at high centrifugal frequencies at which the interface between liquid and air becomes unstable and enables a phase exchange, forwarding the liquid. Burst frequencies of the valve were determined for liquids typically used in biochemical assays: pure water, water with detergent concentrations between 0.01 and 10%, and pure ethanol. Burst frequencies between 8.5 +/- 0.6 and 27.9 +/- 2.0 Hz were measured for different surface tensions. The burst frequencies can be tuned by simple geometrical changes in the valving structure. The valve does not require ultra-precise structures or local surface modifications and is therefore ideal for low-cost microfluidic polymer disks. Potential applications are in the field of multiparameter and panel analysis, such as PCR-genotyping. PMID- 20024044 TI - Precompetitive preclinical ADME/Tox data: set it free on the web to facilitate computational model building and assist drug development. AB - Web-based technologies coupled with a drive for improved communication between scientists have resulted in the proliferation of scientific opinion, data and knowledge at an ever-increasing rate. The increasing array of chemistry-related computer-based resources now available provides chemists with a direct path to the discovery of information, once previously accessed via library services and limited to commercial and costly resources. We propose that preclinical absorption, distribution, metabolism, excretion and toxicity data as well as pharmacokinetic properties from studies published in the literature (which use animal or human tissues in vitro or from in vivo studies) are precompetitive in nature and should be freely available on the web. This could be made possible by curating the literature and patents, data donations from pharmaceutical companies and by expanding the currently freely available ChemSpider database of over 21 million molecules with physicochemical properties. This will require linkage to PubMed, PubChem and Wikipedia as well as other frequently used public databases that are currently used, mining the full text publications to extract the pertinent experimental data. These data will need to be extracted using automated and manual methods, cleaned and then published to the ChemSpider or other database such that it will be freely available to the biomedical research and clinical communities. The value of the data being accessible will improve development of drug molecules with good ADME/Tox properties, facilitate computational model building for these properties and enable researchers to not repeat the failures of past drug discovery studies. PMID- 20024045 TI - An automated microdroplet passive pumping platform for high-speed and packeted microfluidic flow applications. AB - Surface tension driven passive pumping is a microfluidic technology that uses the surface tension present in small droplets to generate flow. To enhance the potential of this type of passive pumping, a new 'micro passive pumping' technique has been developed that allows for high throughput fluidic delivery by combining passive pumping with a small droplet-based fluidic ejection system. Flow rates of up to four milliliters per minute (mL/min) were achieved that are solely limited by the channel geometry and droplet size. Fluid exchange rates can be performed within tens of milliseconds (ms) by delivering fluids from multiple nozzles. The technique can be extended to a multitude of platforms, as channels are not pressurized and therefore do not require bonding to a substrate. This technique provides a novel flow control for high-speed and packeted flow applications without requiring external tubing connections or substrate bonding. PMID- 20024046 TI - Capture of circulating tumor cells from whole blood of prostate cancer patients using geometrically enhanced differential immunocapture (GEDI) and a prostate specific antibody. AB - Geometrically enhanced differential immunocapture (GEDI) and an antibody for prostate-specific membrane antigen (PSMA) are used for high-efficiency and high purity capture of prostate circulating tumor cells from peripheral whole blood samples of castrate-resistant prostate cancer patients. PMID- 20024047 TI - Electrically evoking and electrochemically resolving quantal release on a microchip. AB - A microchip was applied to electrically depolarize rat pheochromocytoma (PC12) cells and to simultaneously detect exocytotic catecholamine release amperometrically. Results demonstrate exocytosis elicited by flowing cells through an electric field generated by a potentiostat circuit in a microchannel, as well as exocytosis triggered by application of an extracellular voltage pulse across. Electrical finite element model (FEM) analysis illustrated that larger cells experienced greater depolarizing excitation from the extracellular electric fields due to the smaller shunt path and higher resistance to current flow in the channel around the cell. Consistent with these simulations, data recorded from cell clusters and large cells exhibited increased release rates relative to data from the smaller cells. Overall, the system was capable of resolving single vesicle quantal release, in the zeptomole range, as well as the kinetics associated with the vesicle fusion process. Analysis of spike population statistics suggested detection of catecholamines from multiple release sites around the cells. The potential for such a device to be used in flow cytometry to evoke and detect exocytosis was demonstrated. PMID- 20024048 TI - A multi-layer microfluidic device for efficient culture and analysis of renal tubular cells. AB - We have developed a simple multi-layer microfluidic device by integrating a polydimethyl siloxane (PDMS) microfluidic channel and a porous membrane substrate to culture and analyze the renal tubular cells. As a model cell type, primary rat inner medullary collecting duct (IMCD) cells were cultured inside the channel. To generate in vivo-like tubular environments for the cells, a fluidic shear stress of 1 dyn/cm(2) was applied for 5 hours, allowing for optimal fluidic conditions for the cultured cells, as verified by enhanced cell polarization, cytoskeletal reorganization, and molecular transport by hormonal stimulations. These results suggest that the microfluidic device presented here is useful for resembling an in vivo renal tubule system and has potential applications in drug screening and advanced tissue engineering. PMID- 20024049 TI - Co-culture of epithelial cells and bacteria for investigating host-pathogen interactions. AB - The human gastrointestinal (GI) tract is a unique environment in which intestinal epithelial cells and non-pathogenic (commensal) bacteria co-exist. This equilibrium is perturbed by the entry of pathogens into the GI tract. A key step in the infection process is the navigation of the pathogen through the commensal bacterial layer to attach to epithelial cells. It has been proposed that the microenvironment that the pathogen encounters in the commensal layer plays a significant role in determining the extent of attachment and colonization. Current culture methods for investigating pathogen colonization are not well suited for investigating this hypothesis as they do not enable co-culture of bacteria and epithelial cells in a manner that mimics the GI tract microenvironment. Here we report the development of a microfluidic co-culture model that enables independent culture of eukaryotic cells and bacteria, and testing the effect of the commensal microenvironment on pathogen colonization. A pneumatically-actuated system was developed to form reversible islands that allow development of bacterial biofilm along with culture of an epithelial cell monolayer. The co-culture model used to develop a commensal Escherichia coli biofilm among HeLa cells, followed by introduction of enterohemorrhagic E. coli (EHEC) into the commensal island, in a sequence that mimics the sequence of events in GI tract infection. Using wild-type E. coli and a tnaA mutant (lacks the signal indole) as the commensal bacteria, we demonstrate that the commensal biofilm microenvironment is a key determinant of EHEC infectivity and virulence. Our model has the potential to be used in fundamental studies investigating the effect of GI tract signals on EHEC virulence as well as for screening of different probiotic strains for modulating pathogen infectivity in the GI tract. PMID- 20024050 TI - Perfused multiwell plate for 3D liver tissue engineering. AB - In vitro models that capture the complexity of in vivo tissue and organ behaviors in a scalable and easy-to-use format are desirable for drug discovery. To address this, we have developed a bioreactor that fosters maintenance of 3D tissue cultures under constant perfusion and we have integrated multiple bioreactors into an array in a multiwell plate format. All bioreactors are fluidically isolated from each other. Each bioreactor in the array contains a scaffold that supports formation of hundreds of 3D microscale tissue units. The tissue units are perfused with cell culture medium circulated within the bioreactor by integrated pneumatic diaphragm micropumps. Electronic controls for the pumps are kept outside the incubator and connected to the perfused multiwell by pneumatic lines. The docking design and open-well bioreactor layout make handling perfused multiwell plates similar to using standard multiwell tissue culture plates. A model of oxygen consumption and transport in the circulating culture medium was used to predict appropriate operating parameters for primary liver cultures. Oxygen concentrations at key locations in the system were then measured as a function of flow rate and time after initiation of culture to determine oxygen consumption rates. After seven days of culture, tissue formed from cells seeded in the perfused multiwell reactor remained functionally viable as assessed by immunostaining for hepatocyte and liver sinusoidal endothelial cell (LSEC) phenotypic markers. PMID- 20024051 TI - In situ generation of pH gradients in microfluidic devices for biofabrication of freestanding, semi-permeable chitosan membranes. AB - We report the in situ generation of pH gradients in microfluidic devices for biofabrication of freestanding, semi-permeable chitosan membranes. The pH-stimuli responsive polysaccharide chitosan was enlisted to form a freestanding hydrophilic membrane structure in microfluidic networks where pH gradients are generated at the converging interface between a slightly acidic chitosan solution and a slightly basic buffer solution. A simple and effective pumping strategy was devised to realize a stable flow interface thereby generating a stable, well controlled and localized pH gradient. Chitosan molecules were deprotonated at the flow interface, causing gelation and solidification of a freestanding chitosan membrane from a nucleation point at the junction of two converging flow streams to an anchoring point where the two flow streams diverge to two output channels. The fabricated chitosan membranes were about 30-60 microm thick and uniform throughout the flow interface inside the microchannels. A T-shaped membrane formed by sequentially fabricating orthogonal membranes demonstrates flexibility of the assembly process. The membranes are permeable to aqueous solutions and are removed by mildly acidic solutions. Permeability tests suggested that the membrane pore size was a few nanometres, i.e., the size range of antibodies. Building on the widely reported use of chitosan as a soft interconnect for biological components and microfabricated devices and the broad applications of membrane functionalities in microsystems, we believe that the facile, rapid biofabrication of freestanding chitosan membranes can be applied to many biochemical, bioanalytical, biosensing applications and cellular studies. PMID- 20024052 TI - Fabrication of a cyclic olefin copolymer planar waveguide embedded in a multi channel poly(methyl methacrylate) fluidic chip for evanescence excitation. AB - The fabrication and characterization of a novel cyclic olefin copolymer (COC) waveguide embedded in a poly(methyl methacrylate), PMMA, fluidic chip configured in a multi-channel format with an integrated monolithic prism for evanescent fluorescence excitation are reported. The fabrication approach allowed the embedded waveguide to be situated orthogonal to a series of fluidic channels within the PMMA wafer to sample fluorescent solutions in these channels using the evanescence properties of the waveguide. Construction of the device was achieved using several fabrication techniques including high precision micromilling, hot embossing and stenciling of a polymer melt to form the waveguide and coupling prism. A waveguide channel was fabricated in the fluidic chip's cover plate, also made from PMMA, and was loaded with a COC solution using a pre-cast poly(dimethylsiloxane), PDMS, stencil containing a prism-shaped recess. The PMMA substrate contained multiple channels (100 microm wide x 30 microm deep with a pitch of 100 microm) that were situated orthogonal to the waveguide to allow penetration of the evanescent field into the sampling solution. The optical properties of the waveguide in terms of its transmission properties and penetration depth of the evanescent field in the adjacent solution were evaluated. Finally, the device was used for laser-induced fluorescence evanescent excitation of a dye solution hydrodynamically flowing through multiple microfluidic channels in the chip and processed using a microscope equipped with a charge-coupled device (CCD) for parallel readout. The device and optical system were able to image 11 channels simultaneously with a limit-of-detection of 7.1 x 10(-20) mol at a signal-to-noise ratio of 2. The waveguide was simple to manufacture and could be scaled to illuminate much higher channel numbers making it appropriate for high-throughput measurements using evanescent excitation. PMID- 20024053 TI - Low-power microfluidic electro-hydraulic pump (EHP). AB - Low-power electrolysis-based microfluidic pumps utilizing the principle of hydraulics, integrated with microfluidic channels in polydimethylsiloxane (PDMS) substrates, are presented. The electro-hydraulic pumps (EHPs), consisting of electrolytic, hydraulic and fluidic chambers, were investigated using two types of electrodes: stainless steel for larger volumes and annealed gold electrodes for smaller-scale devices. Using a hydraulic fluid chamber and a thin flexible PDMS membrane, this novel prototype successfully separates the reagent fluid from the electrolytic fluid, which is particularly important for biological and chemical applications. The hydraulic advantage of the EHP device arises from the precise control of flow rate by changing the electrolytic pressure generated, independent of the volume of the reagent chamber, mimicking the function of a hydraulic press. Since the reservoirs are pre-filled with reagents and sealed prior to testing, external fluid coupling is minimized. The stainless steel electrode EHPs were manufactured with varying chamber volume ratios (1 : 1 to 1 : 3) as a proof-of-concept, and exhibited flow rates of 1.25 to 30 microl/min with electrolysis-based actuation at 2.5 to 10 V(DC). The miniaturized gold electrode EHPs were manufactured with 3 mm diameters and 1 : 1 chamber volume ratios, and produced flow rates of 1.24 to 7.00 microl/min at 2.5 to 10 V(AC), with a higher maximum sustained pressure of 343 KPa, suggesting greater device robustness using methods compatible with microfabrication. The proposed technology is low-cost, low-power and disposable, with a high level of reproducibility, allowing for ease of fabrication and integration into existing microfluidic lab-on-a-chip and analysis systems. PMID- 20024054 TI - Ultrafast high-pressure AC electro-osmotic pumps for portable biomedical microfluidics. AB - This paper details the development of an integrated AC electro-osmotic (ACEO) microfluidic pump for dilute electrolytes consisting of a long serpentine microchannel lined with three dimensional (3D) stepped electrode arrays. Using low AC voltage (1 V rms, 1 kHz), power (5 mW) and current (4.5 mA) in water, the pump is capable of generating a 1.3 kPa head pressure, a 100-fold increase over prior ACEO pumps, and a 1.3 mm/s effective slip velocity over the electrodes without flow reversal. The integrated ACEO pump can utilize low ionic strength solutions such as distilled water as the working solution to pump physiological strength (100 mM) biological solutions in separate microfluidic devices, with potential applications in portable or implantable biomedical microfluidic devices. As a proof-of-concept experiment, the use of the ACEO pumps for DNA hybridization in a microfluidic microarray is demonstrated. PMID- 20024055 TI - A nano-needle/microtubule composite gliding on a kinesin-coated surface for target molecule transport. AB - An alternative method of micro/nano-transport has been achieved by using motor proteins. Microtubules on a kinesin-coated surface have potential to act as a nano-transport system. When microtubules are used as carriers, either cargo or cargo linkers are attached on the microtubule surface. Such cargo attachments can significantly affect kinesin motion. To deal with the difficulty caused by molecular attachment to the microtubule surface, the cargo loading and transport mechanism should be separated. In this work, we propose to use micromachined needles as cargo carriers which then can be transported on microtubules. Because of the separation of needle functionalization and transport mechanism, functionalization of the needles can proceed without any effect on the microtubule structure, significantly increasing the possible types of cargo. We have fabricated silicon needles in mass numbers using a simple and effective method and have shown that the microtubule-needle composites are transported without affecting the kinesin activity. PMID- 20024056 TI - Micro OS-ELISA: Rapid noncompetitive detection of a small biomarker peptide by open-sandwich enzyme-linked immunosorbent assay (OS-ELISA) integrated into microfluidic device. AB - A novel detection system that combines the merits of open-sandwich (OS) enzyme linked immunoadsorbent assay (ELISA) and a microfluidic sensor chip system, and which enables rapid and noncompetitive immunodetection of small antigens of less than 1000 in molecular weight, has been proposed. Equipped with a sensitive thermal lens microscope, a minute amount of the carboxyl-terminal peptide of human osteocalcin (BGP), a biomarker for bone metabolism, was quantified utilizing antigen-dependent stabilization of an antibody variable region (OS principle). In a short analysis time (approximately 12 min), we could attain a detection limit comparable to that of the microplate-based OS ELISA (1 microg L( 1)). In addition, the effects of several pretreatments for serum-derived samples were investigated: an albumin absorption resin, addition of a protease inhibitor cocktail and heat treatment. Each pretreatment was found to be effective. Consequently, intrinsic BGP and its fragments could be detected in healthy human serum with a superior detection limit and working range compared to those of the conventional competitive ELISA method. PMID- 20024057 TI - A low power, on demand electrothermal valve for wireless drug delivery applications. AB - We present a low power, on demand Parylene MEMS electrothermal valve. A novel Omega-shaped thermal resistive element requires low power (approximately mW) and enables rapid valve opening (approximately ms). Using both finite element analysis and valve opening experiments, a robust resistive element design for improved valve opening performance in water was obtained. In addition, a thermistor, as an inrush current limiter, was added into the valve circuit to provide variable current ramping. Wireless activation of the valve using RF inductive power transfer was demonstrated. PMID- 20024058 TI - Integrated active mixing and biosensing using surface acoustic waves (SAW) and surface plasmon resonance (SPR) on a common substrate. AB - This article presents a device incorporating surface plasmon resonance (SPR) sensing and surface acoustic wave (SAW) actuation integrated onto a common LiNbO(3) piezoelectric substrate. The device uses Rayleigh-type SAW to provide active microfluidic mixing in the fluid above the SPR sensor. Validation experiments show that SAW-induced microfluidic mixing results in accelerated binding kinetics of an avidin-biotin assay. Results also show that, though SAW action causes a parasitic SPR response due to heat injection into the fluid, a relatively brief relaxation time following the SAW pulses allows the effect to dissipate, without affecting the overall assay response. Since both SPR sensors and SAW transducers can be fabricated simultaneously using low-cost microfabrication methods on a single substrate, the proposed design is well suited to lab-on-chip applications. PMID- 20024060 TI - Microfluidic probe: a new tool for integrating microfluidic environments and electronic wafer-probing. AB - We demonstrate a new tool for integrating microfluidic channels with commonly used electronic probing techniques. The "microfluidic probe" allows rapid and repeatable fluidic and electronic addressing of small die sites on a variety of substrate types without the need for permanent modification or dicing of the device wafers. We also use the probe to demonstrate locally patterned chemical modification of a substrate. The probes are easily fabricated using standard soft lithography and basic machining making this a widely accessible technique for electronics and fluidics researchers. PMID- 20024059 TI - A new method for studying gradient-induced neutrophil desensitization based on an open microfluidic chamber. AB - During inflammation neutrophils rapidly migrate to the site of tissue damage or infection by following complex gradients of bacterial peptides and host-derived chemokines. The efficiency and speed of neutrophil migration is critically dependent upon the ability of neutrophils to sense new gradients and utilize only those that provide the most direct path to the damaged or infected site. Receptor desensitization plays an important role in migration efficiency and is most commonly studied using bath application of chemotactic factor solutions instead of presenting cells with gradients analogous to those they would experience in vivo. Here we describe a new method for examining gradient-induced neutrophil desensitization using a previously-developed open-chamber microfluidic gradient generator. PMID- 20024061 TI - Molecular BioSystems--celebrations, growth and development. PMID- 20024063 TI - Transcriptional control of the proliferation cluster by the tumor suppressor p53. AB - When genome-wide expression profiles of tumors are compared to those of normal tissues, the most recurring transcriptional pattern is characterized by an increased expression of cell-cycle and proliferation-associated genes, collectively referred to as the 'proliferation cluster'. Tumors with increased expression of the proliferation cluster are frequently associated with augmented proliferation rate, chromosomal instability and metastasis as well as with poor prognosis. Recent in vitro and in vivo data establish a link between the tumor suppressor p53 and the proliferation cluster, implicating loss of p53 activity as the major event responsible for elevated expression of the proliferation cluster in tumors. Moreover, a complex regulatory network, which links p53 with the transcription factors that govern the expression of the proliferation cluster genes, is being gradually elucidated. PMID- 20024064 TI - Of proteins and DNA--proteomic role in the field of chromatin research. AB - To paraphrase Robert Burns's poem To a Mouse, the best laid schemes of DNA protein complex purification often go awry. Chromatin with its heterogeneous and dynamic protein composition remains difficult to analyze. Still critical progress has been made in recent years in characterizing the interface between DNA and proteins due, in part, to significant advances in proteomic technologies. Proteomics has progressed to a point where affinity purification of soluble complexes and protein identification by mass spectrometry are routine. The new challenge for chromatin proteomics lies in studying proteins and protein complexes in their native environment, which is on chromatin. These novel types of data represent an additional layer of information that can be used to better characterize and understand cellular processes. This review will focus on the past contributions as well as on emerging mass spectrometry-based methodologies attempting to better define the complex relationship between proteins, protein complexes and DNA. PMID- 20024065 TI - SCOP/PHLPP and its functional role in the brain. AB - SCOP (suprachiasmatic nucleus (SCN) circadian oscillatory protein) was originally identified in 1999 in a differential display screen of the rat SCN for genes whose expression were regulated in a circadian manner (K. Shimizu, M. Okada, A. Takano and K. Nagai, FEBS Lett., 1999, 458, 363-369). The SCN is the principle pacemaker of the circadian clock, and expression of SCOP protein in the SCN was found to oscillate, increasing during the subjective night, even when animals were housed in constant darkness. SCOP interacts with and inhibits multiple proteins important for intracellular signaling, either by directly binding to K Ras or by dephosphorylating p-Akt and p-PKC. Since the functions of K-Ras, Akt, and PKC are considerably divergent, SCOP may have several roles. We recently discovered that SCOP participates in the formation of long-term hippocampus dependent memories, and other investigators have examined its role in cell proliferation and survival. In this review, we introduce SCOP from its molecular structure to its physiological functions, focusing mainly on its role in ERK1/2 activation and memory consolidation. PMID- 20024066 TI - Small-molecule modulators of the Sonic Hedgehog signaling pathway. AB - Sonic hedgehog (Shh) is the most widely characterized of the three vertebrate Hedgehog homologs, and is essential for proper embryonic development. Shh binds to its receptor, Patched (Ptch1), resulting in the de-repression of Smoothened (Smo). This leads to the activation of Gli2, which regulates the transcription of target genes that include Gli1 and Ptch1. Several synthetic and naturally occurring small-molecule modulators of Smo have been discovered. Shh-signaling antagonists that bind to Smo include cyclopamine, SANT1, and Cur-61414. Shh signaling agonists that bind to Smo include the synthetic small molecules purmorphamine and SAG. Small molecules that inhibit Shh signaling downstream of Smo, GANT58 and GANT61 have also been reported. Robotnikinin inhibits the Shh pathway by directly targeting Shh. Although progress has been made in understanding and modulating Shh signaling, fundamental aspects of Shh signal transduction remain obscure, including the mechanism(s) whereby Ptch1 regulates Smo activity. Small-molecule modulators of Shh signaling provide a means to regulate the activity of a pathway implicated in medulloblastoma, basal cell carcinoma (BCC), pancreatic cancer, prostate cancer and developmental disorders. Several Shh inhibitors have not succeeded in the clinic for unknown reasons, but clinical trials in BCC and pancreatic cancer with the promising Smo antagonists GDC-0449 and IPI-926 are currently underway. PMID- 20024067 TI - Predicting protein-protein interactions in the context of protein evolution. AB - Here we review the methods for the prediction of protein interactions and the ideas in protein evolution that relate to them. The evolutionary assumptions implicit in many of the protein interaction prediction methods are elucidated. We draw attention to the caution needed in deploying certain evolutionary assumptions, in particular cross-organism transfer of interactions by sequence homology, and discuss the known issues in deriving interaction predictions from evidence of co-evolution. We also conject that there is evolutionary knowledge yet to be exploited in the prediction of interactions, in particular the heterogeneity of interactions, the increasing availability of interaction data from multiple species, and the models of protein interaction network growth. PMID- 20024068 TI - Incorporation of unnatural amino acids for synthetic biology. AB - The challenge of synthetic biology lies in the construction of artificial cellular systems. This requires the development of modular "parts" that can be integrated into living systems to elicit an artificial, yet programmed, response or function. The development of methods to engineer proteins bearing unnatural amino acids (UAAs) provides essential components that may address this challenge. Here we review the emerging strategies for incorporating UAAs into proteins with the endgame of engineering artificial cells and organisms. PMID- 20024069 TI - Molecular recognition of poly(A) targeting by protoberberine alkaloids: in vitro biophysical studies and biological perspectives. AB - The use of small molecules to specifically control important cellular functions through binding to nucleic acids is an area of major current interest at the interface of chemical biology and medicinal chemistry. The polyadenylic acid [poly(A)] tail of mRNA has been recently established as a potential drug target due to its significant role in the initiation of translation, maturation and stability of mRNA as well as in the production of alternate proteins in eukaryotic cells. Very recently some small molecule alkaloids of the isoquinoline group have been found to bind poly(A) with remarkably high affinity leading to self-structure formation. Plant alkaloids are small molecules known to have important traditional roles in medicinal chemistry due to their extensive biological activity. Especially, noteworthy are the protoberberine alkaloids that are widely distributed in several botanical families exhibiting myriad therapeutic applications. This review focuses on the structural and biological significance of poly(A) and interaction of protoberberine alkaloids with this RNA structure for the development of new small molecule alkaloids targeted to poly(A) structures as futuristic therapeutic agents. PMID- 20024070 TI - Biomolecule detection with peroxidase-mimicking DNAzymes; expanding detection modality with fluorogenic compounds. AB - The identification of a suitable non-fluorescent molecule that can be oxidized by G-quadruplexes into a fluorescent product will be important for several fields such as bioanalyte sensing and cancer therapeutic discovery. Herein, we demonstrate that 2',7'-dichlorodihydrofluorescein diacetate is a superior reducing substrate for the fluorometric detection of bioanalytes using peroxidase mimicking G-quadruplex DNAzymes. PMID- 20024071 TI - Rapid identification of orexin receptor binding ligands using cell-based screening accelerated with magnetic beads. AB - We report here a simple and rapid method by which to screen one bead one compound libraries for highly specific ligands to cell surface proteins such as G protein coupled receptors. This protocol, which harvests "hits" in a cell-based binding screen magnetically, eliminates the most tedious aspects of previously published bead screening techniques and allows millions of different compounds to be screened rapidly and cheaply. The method is demonstrated using the orexin receptor 1, which resulted in the isolation of moderate potency antagonists. PMID- 20024072 TI - Methodological considerations in the development of HPLC-MS methods for the analysis of rodent plasma for metabonomic studies. AB - A study of the factors involved in obtaining valid global metabolite profiles from the HPLC-MS of rat or mouse plasma for the purposes of metabonomic analysis has been undertaken. Plasma proteins were precipitated with three volumes of either methanol or acetonitrile. Chromatographic separations were performed on a C18-bonded stationary phase using 3.5 and 5 mum particles packed into 2.1 and 4.6 mm i.d. formats, respectively, and on a C8 phase using 3.5 mum particles and a 2.1 mm i.d. column. Three reversed-phase gradient solvent systems, based on acidified water-acetonitrile, acidified water-methanol and acidified water methanol-acetonitrile mixtures, were investigated. The column eluent was analysed with both positive and negative electrospray ionisation using a quadrupole-linear ion trap mass spectrometer. These studies revealed that while accurate classification of sample type can be made, there are a number of methodological problems associated with the analysis of plasma with respect to factors such as repeatability and column longevity. In particular, special care has to be taken to ensure that the analytical system is properly "conditioned" by the repeated injection of matrix samples. The use of biological quality control (QC) samples provided an important means of monitoring method performance. Finally, the source of the plasma (Zucker wild-type or (fa/fa) rat or mouse tumour model) also appeared to have an effect on the repeatability of the methodology. PMID- 20024073 TI - In vivo proton HR-MAS NMR metabolic profile of the freshwater cladoceran Daphnia magna. AB - The method concerning in vivo proton HR-MAS NMR metabolic profiling of the freshwater cladoceran Daphnia magna is presented. Viability tests of D. magna under different spinning rates were performed. All surviving daphnids after analysis have developed eggs and embryos like control animals. Better survival rate at the slowest rotation speed were observed. The maximum length of analysis during which the integrity of the daphnid is maintained was assessed. The recorded proton spectra of in vivo daphnia were attributed to lipids from the triglycerol category. Saturated and unsaturated omega-3 like fatty acid moieties of triacylglycerol were well identified. The relationship between physiological state of daphnids and lipid profile are discussed. PMID- 20024074 TI - Competitively selected protein ligands pay their increase in specificity by a decrease in affinity. AB - Protein-ligand interactions characterise and govern the current state and fate of a living cell. The specificity of proteins is mainly determined by the relative affinities to each potential ligand. To investigate the consequences and potentials of ligands with increased specificity in comparison with ligands optimised solely for affinity, it was necessary to identify ligands that are optimised towards specificity instead of a barely optimised affinity to a given target. In the presented example, a modified phage display screening procedure yielded specific ligands for the LckSH3 domain. We found that increased specificity of one of the hereby obtained ligands for LckSH3 is achieved at the cost of a slightly reduced affinity to LckSH3 and a drastically reduced affinity to other SH3 domains. A surface plasmon resonance experiment simulating in vivo like realistic competitive binding conditions exerted enhanced binding behaviour of the specific ligand under these binding conditions. The experimental data, together with a mathematical model describing the complex experimental situation, and theoretical considerations lead to the conclusion that increased specificity is achieved at the cost of reduced affinity, but after all, it pays if the ligand is applied under realistic, i.e. competitive, conditions. PMID- 20024075 TI - Transcriptional control of the quorum sensing response in yeast. AB - Quorum sensing is a process of intercellular communication. It allows individual cells to assess population density and to co-ordinate behaviour by secreting and sensing communication molecules. In the yeast Saccharomyces cerevisiae, the communication molecules are the aromatic alcohols tryptophol and phenylethanol, and quorum sensing regulates the transition between the solitary yeast form and the filamentous form. Though it is known that addition of these communication molecules to yeast cultures causes large changes in gene expression, how these changes are orchestrated and whether this system is conserved in related fungal species is still unknown. In this work, by employing an integrated computational approach that makes use of large-scale genomics datasets, such as ChIP-ChIP and expression analysis upon deletion and over-expression of transcriptional factors, we predict CAT8 and MIG1 as key transcriptional regulators that control the differential expression of the genes affected by aromatic alcohol communication. In addition, through a comparative genomic analysis involving 31 fungal species, we show that the S. cerevisiae quorum sensing system is a recent evolutionary innovation and that the genes which are differentially expressed upon treatment with these molecules are distributed across the genome in a highly non-random manner. The identified transcription factors will aid in further unravelling the molecular mechanisms of S. cerevisiae quorum sensing and may facilitate the engineering of regulatory circuits for applications such as the expression of heterologous proteins via aromatic alcohols. PMID- 20024076 TI - Amino acid transport in thermophiles: characterization of an arginine-binding protein in Thermotoga maritima. AB - Members of the periplasmic binding protein superfamily are involved in the selective passage of ligands through bacterial cell membranes. The hyperthermophilic eubacterium Thermotoga maritima was found to encode a highly stable and specific periplasmic arginine-binding protein (TM0593). Following signal sequence removal and overexpression in Escherichia coli, TM0593 was purified by thermoprecipitation and affinity chromatography. The ultra-stable protein with a monomeric molecular weight of 27.7 kDa was found to exist as both a homodimer and homotrimer at appreciable concentrations even under strongly denaturing conditions, with an estimated transition temperature of 116 degrees C. Its multimeric structure may provide further evidence of the importance of quaternary structure in the movement of nutrients across bacterial membranes. Purified and refolded TM0593 was further characterized by fluorescence spectroscopy, mass spectrometry, and circular dichroism to demonstrate the specificity of the protein for arginine and to elucidate structural changes associated with arginine binding. The protein binds arginine with a dissociation constant of 20 muM as determined by surface plasmon resonance measurements. Due to its high thermodynamic stability, TM0593 may serve as a scaffold for the creation of a robust fluorescent biosensor. PMID- 20024077 TI - Genome-scale modeling and in silico analysis of mouse cell metabolic network. AB - Genome-scale metabolic modeling has been successfully applied to a multitude of microbial systems, thus improving our understanding of their cellular metabolisms. Nevertheless, only a handful of works have been done for describing mammalian cells, particularly mouse, which is one of the important model organisms, providing various opportunities for both biomedical research and biotechnological applications. Presented herein is a genome-scale mouse metabolic model that was systematically reconstructed by improving and expanding the previous generic model based on integrated biochemical and genomic data of Mus musculus. The key features of the updated model include additional information on gene-protein-reaction association, and improved network connectivity through lipid, amino acid, carbohydrate and nucleotide biosynthetic pathways. After examining the model predictability both quantitatively and qualitatively using constraints-based flux analysis, the structural and functional characteristics of the mouse metabolism were investigated by evaluating network statistics/centrality, gene/metabolite essentiality and their correlation. The results revealed that overall mouse metabolic network is topologically dominated by highly connected and bridging metabolites, and functionally by lipid metabolism that most of essential genes and metabolites are from. The current in silico mouse model can be exploited for understanding and characterizing the cellular physiology, identifying potential cell engineering targets for the enhanced production of recombinant proteins and developing diseased state models for drug targeting. PMID- 20024078 TI - Cross-species analysis of the glycolytic pathway by comparison of molecular interaction fields. AB - The electrostatic potential of an enzyme is a key determinant of its substrate interactions and catalytic turnover. Here we invoke comparative analysis of protein electrostatic potentials, along with sequence and structural analysis, to classify and characterize all the enzymes in an entire pathway across a set of different organisms. The electrostatic potentials of the enzymes from the glycolytic pathway of 11 eukaryotes were analyzed by qPIPSA (quantitative protein interaction property similarity analysis). The comparison allows the functional assignment of neuron-specific isoforms of triosephosphate isomerase from zebrafish, the identification of unusual protein surface interaction properties of the mosquito glucose-6-phosphate isomerase and the functional annotation of ATP-dependent phosphofructokinases and cofactor-dependent phosphoglycerate mutases from plants. We here show that plants possess two parallel pathways to convert glucose. One is similar to glycolysis in humans, the other is specialized to let plants adapt to their environmental conditions. We use differences in electrostatic potentials to estimate kinetic parameters for the triosephosphate isomerases from nine species for which published parameters are not available. Along the core glycolytic pathway, phosphoglycerate mutase displays the most conserved electrostatic potential. The largest cross-species variations are found for glucose-6-phosphate isomerase, enolase and fructose-1,6-bisphosphate aldolase. The extent of conservation of electrostatic potentials along the pathway is consistent with the absence of a single rate-limiting step in glycolysis. PMID- 20024079 TI - High-confidence mapping of chemical compounds and protein complexes reveals novel aspects of chemical stress response in yeast. AB - Chemical genetics in yeast has shown great potential for clarifying the pharmacology of various drugs. Investigating these results from a systems perspective has uncovered many facets of natural chemical tolerance, but many cellular interactions of chemicals still remain poorly understood. To uncover previously overlooked players in resistance to chemical stress we integrated several independent chemical genetics datasets with protein-protein interactions and a comprehensive collection of yeast protein complexes. As a consequence we were able to identify the potential targets and mode of action of certain poorly understood compounds. However, most complexes recovered in our analysis appear to perform indirect roles in countering deleterious effects of chemicals by constituting an underlying intricate buffering system that has been so far under appreciated. This buffering role appears to be largely contributed to by complexes pertaining to chromatin and vesicular dynamics. The former set of complexes seems to act by setting up or maintaining gene expression states necessary to protect the cell against chemical effects. Among the latter complexes we found an important role for specific vesicle tethering complexes in tolerating particular sets of compounds, indicating that different chemicals might be routed via different points in the intracellular trafficking system. We also suggest a general operational similarity between these complexes and molecular capacitors (e.g. the chaperone Hsp90). Both have a key role in increasing the system's robustness, although at different levels, through buffering stress and mutation, respectively. It is therefore conceivable that some of these complexes identified here might have roles in molding the evolution of chemical resistance and response. PMID- 20024080 TI - Metabolic profiling of the fission yeast S. pombe: quantification of compounds under different temperatures and genetic perturbation. AB - Metabolomics is a rapidly growing branch of post-genomic chemical biology. The fission yeast Schizosaccharomyces pombe is an excellent eukaryotic model organism. Although the entire S. pombe genome has been sequenced and detailed transcriptomic analyses were performed, little metabolic profiling has been done. Here we report the first global semi-quantitative analysis of the S. pombe metabolome using liquid chromatography high-resolution mass spectrometry. Procedures to obtain metabolic compounds from S. pombe extracts were established. One hundred and twenty-three distinct metabolites were identified while approximately 1900 peaks from the approximately 6000 observed were assigned. A software system (MZviewer) was developed to visualize semi-quantitative metabolome data using a dynamically generated scatter plot. We examined the metabolome of S. pombe cells exponentially grown in synthetic culture medium (EMM2) at two different temperatures, 26 degrees C and 36 degrees C. The profiles were similar except for varying amounts of certain amino acids and a significant increase in several compounds at 36 degrees C, such as trehalose (200-fold), glycerophosphoethanolamine (50-fold), arabitol (16-fold), ribulose (8-fold), and ophthalmic acid (5-fold). Reproducibility was demonstrated using a deletion mutant sib1Delta that lacked ferrichrome synthetase and showed no significant metabolic effects except the disappearance of the hexapeptide ferrichrome and the appearance of a putative dipeptide precursor. Taking advantage of the metabolic profile similarity at 26 degrees C and 36 degrees C, we analyzed the metabolome of a temperature-sensitive hcs1-143 mutant defective in the HMG-CoA synthase. As expected, HMG-CoA was decreased. In addition, extensive secondary metabolic effects, including a decrease in urea cycle intermediates and an increase in acetylated compounds, were observed. These findings confirm that S. pombe can be applied as an appropriate model to monitor metabolic responses to environmental conditions as well as genetic perturbations. PMID- 20024081 TI - Alternate dab-aegPNAs: synthesis, nucleic acid binding studies and biological activity. AB - As part of our research on new oligonucleotide analogs for therapeutic and diagnostic use, here we explored the ability of an alternate dab-aegPNA oligomer to bind complementary natural nucleic acids. The alternate homothymine dab aegPNA, synthesized following a chirally safe procedure and fully characterized by ESIMS and CD, was capable of forming hybrids with complementary DNA and RNA with enhanced thermal stability in comparison to natural oligomers, as shown by CD and UV spectroscopies. The stoichiometry of the complexes formed was determined by CD titration experiments that suggested triple helices formation. With respect to an analogous t(12) strand composed entirely of aegPNA, the chiral alternate t(12) oligomer presented an enhanced solubility in aqueous medium and did not form aggregates. Human serum stability assays performed on the new alternate oligomer evidenced a noteworthy enzymatic resistance. Moreover, the efficiency of dab-aegPNA in interfering with the reverse transcription of eukaryotic mRNA, and the absence of cytotoxic effects of the new analog were demonstrated, encouraging us to further study this chiral PNA analog in view of its possible in vivo/in vitro biotechnological applications. PMID- 20024082 TI - Selective oxidative demethylation of veratric acid to vanillic acid by CYP199A4 from Rhodopseudomonas palustris HaA2. AB - CYP199A4 (RPB3613) from Rhodopseudomonas palustris HaA2 is a heme monooxygenase that catalyzes the hydroxylation of para-substituted benzoic acids. Monooxygenase activity of CYP199A4 can be reconstituted in a Class I electron transfer chain with an associated [2Fe-2S] ferredoxin, HaPux, (RPB3614) and the flavin-dependent reductase, HaPuR, (RPB3656) that is not associated with a CYP gene. CYP199A4 and the ferredoxin HaPux are produced in greater quantities using recombinant Escherichia coli expression systems when compared to the equivalent proteins in the closely related CYP199A2-Pux-PuR Class I system from R. palustris CGA009. HaPuR and HaPux can also replace PuR and Pux in supporting the CYP199A2 enzyme turnover with high activity. Whole-cell in vivo substrate oxidation systems for CYP199A4 and CYP199A2 with HaPux and HaPuR as the electron transfer proteins have been constructed. These E. coli systems were capable of selectively demethylating veratric acid at the para position to produce vanillic acid at rates of up to 15.3 microM (g-cdw)(-1) min(-1) and yields of up to 1.2 g L(-1). PMID- 20024083 TI - The influence of EDTA and citrate anticoagulant addition to human plasma on information recovery from NMR-based metabolic profiling studies. AB - The widely-used blood anticoagulants citrate and EDTA give rise to prominent peaks in (1)H NMR spectra of plasma samples collected in epidemiological and clinical studies, and these cause varying levels of interference in recovering biochemical information on endogenous metabolites. To investigate both the potential metabolic information loss caused by these substances and any possible inter-molecular interactions between the anticoagulants and endogenous components, the (1)H NMR spectra of 40 split human plasma samples collected from 20 individuals into either citrate or EDTA have been analysed. Endogenous metabolite peaks were selectively obscured by large citrate peaks or those from free EDTA and its calcium and magnesium complexes. It is shown that the endogenous metabolites that give rise to peaks obscured by those from EDTA or citrate almost invariably also have other resonances that allow their identification and potential quantitation. Also, metabolic information recovery could be maximised by use of spectral editing techniques such as spin-echo, diffusion-editing and J-resolved experiments. The NMR spectral effects of any interactions between the added citrate or EDTA and endogenous components were found to be negligible. Finally, identification of split samples was feasible using simple multivariate statistical approaches such as principal components analysis. Thus even when legacy epidemiological plasma samples have been collected using the NMR-inappropriate citrate or EDTA anticoagulants, useful biochemical information can still be recovered effectively. PMID- 20024084 TI - 'On-the-fly' optical encoding of combinatorial peptide libraries for profiling of protease specificity. AB - Large solid-phase combinatorial libraries currently play an important role in areas such as infectious disease biomarker discovery, profiling of protease specificity and anticancer drug discovery. Because compounds on solid support beads are not positionally-encoded as they are in microarrays, innovative methods of encoding are required. There are many advantages associated with optical encoding and several strategies have been described in the literature to combine fluorescence encoding methods with solid-phase library synthesis. We have previously introduced an alternative fluorescence-based encoding method ("colloidal barcoding"), which involves encoding 10-20 mum support beads during a split-and-mix synthesis with smaller 0.6-0.8 mum silica colloids that contain specific and identifiable combinations of fluorescent dye. The power of this 'on the-fly' encoding approach lies in the efficient use of a small number of fluorescent dyes to encode millions of compounds. Described herein, for the first time, is the use of a colloid-barcoded library in a biological assay (i.e., protease profiling) combined with the use of confocal microscopy to decode the colloidal barcode. In this proof-of-concept demonstration, a small focussed peptide library was optically-encoded during a combinatorial synthesis, incubated with a protease (trypsin), analysed by flow cytometry and decoded via confocal microscopy. During assay development, a range of parameters were investigated and optimised, including substrate (or probe) loading, barcode stability, characteristics of the peptide-tagging fluorophore, and spacer group configuration. Through successful decoding of the colloidal barcodes, it was confirmed that specific peptide sequences presenting one or two cleavage sites were recognised by trypsin while peptide sequences not cleavable by trypsin remained intact. PMID- 20024085 TI - Comparison of the interactomic networks of different species in terms of accessibility. AB - Protein-protein interaction networks were investigated in terms of outward accessibility, which quantifies the effectiveness of each protein in accessing other proteins and is related to the internality of nodes. By comparing the accessibility between 144 ortholog proteins in yeast and the fruit fly, we found that the accessibility tends to be higher among proteins in the fly than in yeast. In addition, z-scores of the accessibility calculated for different species revealed that the protein networks of less evolved species tend to be more random than those of more evolved species. The accessibility was also used to identify the border of the yeast protein interaction network, which was found to be mainly composed of viable proteins. PMID- 20024086 TI - A residue outside the active site CXXC motif regulates the catalytic efficiency of Glutaredoxin 3. AB - The glutaredoxin (Grx) family of oxidoreductases has a conserved residue at position 8 that varies between Arginine in Grx1 and Lysine in Grx3. It has been proposed that this Arg/Lys change is the main cause for the 35 mV difference in redox potential between the two enzymes. To gain insights into the catalytic machinery of Grx3 and directly evaluate the role of residue 8 in the catalysis of thiol-disulfide exchange by this enzyme, we synthesized the "wild type" enzyme (sGrx3), and four analogues substituting the lysine at position 8 with arginine, ornithine (Orn), citrulline (Cit) and norvaline (Nva). The redox potential and equilibration kinetics with thioredoxin (Trx1) were determined for each enzyme by fluorescence intensity. While minor effects on redox potential were observed, we found that residue 8 had a more marked effect on the catalytic efficiency of this enzyme. Surprisingly, truncation of the functional group resulted in a more efficient enzyme, Lys8Nva, exhibiting rate constants that are an order of magnitude higher than sGrx3 for both forward and reverse reactions. These observations pose the question why would a residue that reduces the rate of enzyme turnover be evolutionarily conserved? The significant changes in the kinetic parameters suggest that this position plays an important role in the thiol-disulfide exchange reaction by affecting the nucleophilic thiolate through electrostatic or hydrogen bonding interactions. Since the reduced Grx has an exposed thiol that could easily be alkylated, either Arg or Lys could act as a gatekeeper that deters unwanted electrophiles from attacking the active site thiolate. PMID- 20024087 TI - Effect of clustered peptide binding on DNA condensation. AB - DNA condensation in-vitro has been studied as a model system to reveal common principles underlying gene packaging in biology, and as the critical first step towards the development of non-viral gene delivery vectors. In this study, we use a bio-inspired approach, where small DNA-binding peptides are controllably clustered by an amphiphilic block copolymer scaffold, to reveal the effect of clustered peptide binding on the energetics, size, shape and physical properties of DNA condensation in-vitro. This provides insights into the general architectural effect of gene-binding proteins on DNA condensation process. Moreover, the versatility afforded by regulating the clustering density and composition of peptides may provide a novel design platform for gene delivery applications in the future. PMID- 20024088 TI - Poly(methyl methacrylate)-graft-oligoamines as low cytotoxic and efficient nonviral gene vectors. AB - A series of poly(methyl methacrylate)-graft-oligoamines (PMMA-g-oligoamines), including PMMA-g-DETA, PMMA-g-TETA and PMMA-g-TEPA, were synthesized through aminolysis of the PMMA with diethylenetriamine, triethylenetetramine and tetraethylenepentamine. Agarose gel retardation assay indicated that PMMA-g oligoamines had good binding capability with plasmid DNA, and the binding capability increased with increasing length of oligoamines and content of nitrogen (N%). The results of particle size, zeta potential and morphology observation further showed that the PMMA-g-oligoamines could condense DNA efficiently and the PMMA-g-oligoamine/DNA complexes were uniform nanospheres. The in vitro cell viability indicated that PMMA-g-oligoamines were less toxic than 25 kDa PEI, though the cytotoxicity of PMMA-g-oligoamines increased slightly with increasing length of oligoamines as well as the N% of PMMA-g-oligoamines. The transfection efficiency of PMMA-g-oligoamines/DNA complexes in 293 T and HeLa cells demonstrated that PMMA-g-oligoamines could transfect cells efficiently with increasing the length of oligoamines, especially PMMA-g-TEPA with highest N%, and showed similar transfection capability as 25 kDa PEI. The cellular uptake study showed that the distribution of YOYO-1 labeled DNA in the cytoplasm and nuclei increased gradually with increasing length of oligoamines. PMID- 20024089 TI - Dynamical effects of epigenetic silencing of 14-3-3sigma expression. AB - The development and progression of malignant tumours are often due to deregulated cell cycle control involving a plethora of different molecules. Among these, tumour suppressor proteins like p53 play a crucial role. p53 induces 14-3-3sigma, a multifunctional protein kinase inhibitor, centrally involved in cell cycle control and DNA damage repair after genotoxic stress. Recently, it has been shown that 14-3-3sigma is epigenetically silenced in a variety of tumours, which might contribute to tumour development and progression via impaired cell cycle control. In addition, p53, its inhibitor MDM2 and 14-3-3sigma form a signalling module in which 14-3-3sigma positively regulates the activity of p53 through feedback regulation. Here we present a mathematical model integrating the effects of 14-3 3sigma gene silencing, the dynamics of 14-3-3sigma induction and compartmentalisation by genotoxic stress and the role of interacting molecules p53 and MDM2. In vitro experiments with different melanoma cell lines were performed and our mathematical model was subjected to computer simulations to analyse different scenarios of activation depending on gene methylation status and DNA damage levels. Our analysis indicates that 14-3-3sigma expression is silenced by high gene methylation, but also that strong stimulation is necessary to induce 14-3-3sigma expression in cases of intermediate levels of gene methylation. More intriguingly, the model suggests that epigenetic silencing of 14-3-3sigma affects p53 dynamics in a synergistic way, such that the accumulative effect of partial downregulation of p53 expression and reduction of its nuclear fraction could affect drastically the activity of p53 as a transcription factor. PMID- 20024092 TI - New therapeutic potential for psychoactive natural products. AB - Much of our knowledge in neuroscience was discovered through the study of mind altering natural products. However, although much has been learned about human physiology and basic biological processes, the underlying causes of CNS disorders and other disease states are still elusive. Based on its main past successes, the continued study of mind-altering compounds promises to yield novel agents that may be developed into medications and to identify new targets for the treatment of diseases. This Highlight describes the history of investigations into several classes of mind-altering natural products and relates recent and potential therapeutic uses for these agents. PMID- 20024093 TI - Biosynthesis and biomimetic synthesis of alkaloids isolated from plants of the Nitraria and Myrioneuron genera: an unusual lysine-based metabolism. AB - This review describes a wide panel of alkaloids isolated from plants of the Nitraria genus, focusing on their biosynthesis and discussing the resulting biomimetic chemistry in relevant cases. The scope is purposely limited to alkaloids derived at least to some extent from L-lysine, considering that most of these molecules have unique structures and are specific to the genus. Some of the biosynthetic pathways described are taken from the literature, but others are proposed here for the first time. The latter are mostly hypotheses justified by the fact that they are based on metabolic routes frequently encountered for other Nitraria alkaloids, and thus permit unification of the biosynthesis around common pivotal biosynthetic intermediates. Myrioneuron alkaloids are also presented as a newly discovered class with striking similarities to Nitraria alkaloids. PMID- 20024094 TI - Prenylated indole derivatives from fungi: structure diversity, biological activities, biosynthesis and chemoenzymatic synthesis. AB - Prenylated indole alkaloids are hybrid natural products derived from prenyl diphosphates and tryptophan or its precursors and widely distributed in filamentous fungi, especially in the genera Penicillium and Aspergillus of ascomycota. These compounds represent a group of natural products with diverse chemical structures and biological activities. Significant progress on their biosynthesis has been achieved in recent years by identification of biosynthetic gene clusters from genome sequences and by molecular biological and biochemical investigations. In addition, a series of prenylated indole derivatives have been produced by chemoenzymatic synthesis using overproduced and purified enzymes. PMID- 20024091 TI - Epigenome manipulation as a pathway to new natural product scaffolds and their congeners. AB - The covalent modification of chromatin is an important control mechanism used by fungi to modulate the transcription of genes involved in secondary metabolite production. To date, both molecular-based and chemical approaches targeting histone and DNA posttranslational processes have shown great potential for rationally directing the activation and/or suppression of natural-product encoding gene clusters. In this Highlight, the organization of the fungal epigenome is summarized and strategies for manipulating chromatin-related targets are presented. Applications of these techniques are illustrated using several recently published accounts in which chemical-epigenetic methods and mutant studies were successfully employed for the de novo or enhanced production of structurally diverse fungal natural products (e.g., anthraquinones, cladochromes, lunalides, mycotoxins, and nygerones). PMID- 20024095 TI - Triterpenoids. AB - This review covers the isolation and structure determination of triterpenoids including squalene derivatives, protostanes, lanostanes, holostanes, cycloartanes, dammaranes, euphanes, tirucallanes, tetranortriterpenoids, lupanes, oleananes, friedelanes, ursanes, hopanes, isomalabaricanes and saponins; 574 references are cited. PMID- 20024096 TI - Clickable fluorescent dyes for multimodal bioorthogonal imaging. AB - Bioorthogonal ligation with functionalized fluorescent dyes enables visualization of nuclei acids, proteins and metabolites in biological systems. Bright and modular azide- and alkyne-functionalized dyes are therefore needed to expand the fluorescence imaging capabilities of bioorthogonal ligation methods. We describe the concise synthesis of clickable fluorescent dyes based on 2-dicyanomethylene-3 cyano-2,5-dihydrofuran fluorophores and demonstrate their utility for multicolor imaging of azide- and alkyne-modified proteins as well as FRET studies. PMID- 20024097 TI - Towards an asymmetric synthesis of the bacterial peptide deformylase (PDF) inhibitor fumimycin. AB - Studies towards the synthesis of the bacterial peptide deformylase (PDF) inhibitor fumimycin are reported. The synthetic approach features an organocatalytic access to the alpha,alpha-disubstituted amino acid unit and results in the synthesis of an advanced intermediate which already contains all functionalities of fumimycin. PMID- 20024098 TI - Robust preparation of novel imidazo[5,1-b][1,3,4]oxadiazoles. AB - Cyclodehydration of amino acid-derived acyl hydrazide amides to the corresponding oxadiazoles was followed by a second dehydration event, smoothly furnishing the novel imidazo[5,1-b][1,3,4]oxadiazole motif . PMID- 20024099 TI - Total synthesis of (5R,6R,8R,9S)-(-)-5,9Z-indolizidine 221T using sulfinimine derived N-sulfinyl beta-amino ketones. AB - The first total asymmetric synthesis of the poison frog alkaloid (-)-221T, a 5,6,8-trisubstituted indolizidine is described. The key core piperidine ring was constructed via an acid catalyzed intramolecular cascade Mannich cyclization reaction of a N-sulfinyl syn-alpha-methyl beta-amino ketone and crotonaldehyde. The beta-amino ketone was prepared via the reaction of prochiral lithium Weinreb amide enolate with an enantiopure N-2,4,6-triisopropylphenylsulfinyl imine. PMID- 20024100 TI - Novel asymmetrically functionalized bis-dipicolylamine metal complexes: peripheral decoration of a potent anion recognition scaffold. AB - We report the design and synthesis of a novel class of asymmetrically functionalized, ditopic bis-dipicolylamine (BDPA) ligands. A key feature of this research involved the controlled, sequential functional group decoration of a potent molecular recognition scaffold. Calorimetric screening identified a BDPA analogue as a highly potent (K(a) approximately 10(6) M(-1)) and selective sensor for inorganic phosphate. PMID- 20024101 TI - Introduction of disulfide bond to the main chain of PNA to switch its hybridization and invasion activity. AB - In order to facilitate the removal of peptide nucleic acid (PNA), when necessary, from its duplexes and invasion complexes, a disulfide bond was introduced to its main chain. The disulfide bond was readily cleaved by various reducing agents (2 mercaptoethanol, dl-dithiothreitol, and tris(2-carboxyethyl)phosphine) even when the PNA was forming a duplex with its complementary DNA. The resultant two short PNA fragments were spontaneously removed from the DNA. Double-duplex invasion complexes of two disulfide-containing PNA strands were also promptly cleaved by the reducing agents. By using this modified PNA, a desired DNA fragment was picked up from DNA mixtures, and obtained in a pure form (free from the PNA) by the reductive treatment. Importantly, this separation was achieved at low temperatures (e.g., 37 degrees C), where all the DNAs (and other biomolecules if any) should be kept intact. Strong potential of the modified PNA for various biological applications has been indicated. PMID- 20024102 TI - Selective demethylation and debenzylation of aryl ethers by magnesium iodide under solvent-free conditions and its application to the total synthesis of natural products. AB - An efficient selective demethylation and debenzylation method for aryl methyl/benzyl ethers using magnesium iodide under solvent-free conditions has been developed and applied to the synthesis of natural flavone and biphenyl glycosides. A variety of functional groups including glycoside were tolerated under the reaction conditions. Experimental results indicated that the removal of an O-benzyl group was easier than that of an O-methyl group, regardless of wherever they were meta or para to the carbonyl. Thus selective debenzylation can be achieved for substrates bearing both benzyloxy and methoxy groups. PMID- 20024103 TI - An efficient copper-catalyzed synthesis of hexahydro-1H- phenothiazines. AB - Hexahydro-1H-phenothiazine moieties can be synthesized by domino aziridine ring opening with o-halothiophenols followed by copper-catalyzed Goldberg coupling cyclization with good to excellent yields. Less reactive o-chlorothiophenols are also successfully utilized for the domino reaction to synthesize phenothiazine derivatives. This methodology is successfully applied in the synthesis of phenothiazine skeletons containing biologically active molecules such as antihistamine agents. PMID- 20024104 TI - Facile syntheses of (+)-gabosines A, D, and E. AB - (+)-Gabosines A (12), D (4), and E (5), which share the same trihydroxycyclohexenone skeleton, were synthesized from enone 11 as the common intermediate. The key building block 11 was accessed by an intramolecular aldol cyclization of a diketone derived from D-glucose (8). PMID- 20024106 TI - Efficient one-pot synthesis of 2,4-di(het)aryl and 2,4-diamino pyrido[3,2 d]pyrimidines involving regioselective S(N)Ar and palladium-catalyzed reactions. AB - An efficient and original synthesis of various 2,4-disubstituted pyrido[3,2 d]pyrimidines is reported. One-pot di(het)arylation and diamination approaches were used to obtain highly functionalized products in very good yields. The two one-pot processes were compared to their step-by-step related synthesis. Both routes started from 2,4-dichloropyrido[3,2-d]pyrimidine and included a chlorine discrimination during the first reaction. PMID- 20024105 TI - Pyridine and pyrimidine analogs of acetaminophen as inhibitors of lipid peroxidation and cyclooxygenase and lipoxygenase catalysis. AB - Herein we report an investigation of the efficacy of pyridine and pyrimidine analogs of acetaminophen (ApAP) as peroxyl radical-trapping antioxidants and inhibitors of enzyme-catalyzed lipid peroxidation by cyclooxygenases (COX) and lipoxygenases (LOX). In inhibited autoxidations we find that ApAP, the common analgesic and antipyretic agent, is a very good antioxidant with a rate constant for reaction with peroxyl radicals (k(inh) = 5 x 10(5) M(-1) s(-1)) that is higher than many widely-used phenolic antioxidants, such as the ubiquitous butylated hydroxytoluene (BHT). This reactivity is reduced substantially upon incorporation of nitrogen into the phenolic ring, owing to an increase in the O-H bond dissociation enthalpy of pyridinols and pyrimidinols with respect to phenols. Incorporation of nitrogen into the phenolic ring of ApAP was also found to decrease its efficacy as an inhibitor of prostaglandin biosynthesis by ovine COX-1 (oCOX-1). This is explained on the basis of an increase in its oxidation potential and its reduced reactivity as a reducing co-substrate of the peroxidase protoporphyrin. In contrast, the efficacy of ApAP as an inhibitor of lipid hydroperoxide biosynthesis by soybean LOX-1 (sLOX-1) increased upon incorporation of nitrogen into the ring, suggesting a different mechanism of inhibition dependent on the acidity of the phenolic O-H which may involve chelation of the catalytic non-heme iron atom. The greater stability of the 3-pyridinols and 5 pyrimidinols to air oxidation as compared to phenols allowed us to evaluate some electron-rich pyridinols and pyrimidinols as inhibitors of oCOX-1 and sLOX-1. While the pyridinols had the best combination of activities as antioxidants and inhibitors of oCOX-1 and sLOX-1, they were found to be more toxic than ApAP in preliminary assays in human hepatocellular carcinoma (HepG2) cell culture. The pyrimidinols, however, were up to 17-fold more reactive to peroxyl radicals and up to 25-fold better inhibitors of prostaglandin biosynthesis than ApAP, with similar cytotoxicities to HepG2 cells at high levels of exposure. PMID- 20024107 TI - Free-energy relationships for the interactions of tryptophan with phosphocholines. AB - In membrane proteins and peptides, tryptophan exhibits a marked tendency to occur in locations that correspond to the interfacial region of the lipid bilayer. The relative contributions of electrostatic, dipolar, hydrophobic and conformational effects on the interactions of tryptophan with lipids have been the subject of much speculation. In order to elucidate the fundamental properties of tryptophan phosphocholine interactions in the absence of competing factors such as protein conformation and membrane perturbation, we have determined the binding characteristics of a homologous series of tryptophan analogues to 1,2-dimyristoyl sn-glycero-3-phosphocholine (DMPC) in deuterochloroform using NMR titrimetric approaches. The data are analysed using a binding model that includes lipid aggregation and the explicit association of water with the lipid. For a series of substituents (OMe, Me, H, F, Cl, Br, I, NO(2)) at the 5-position of the indole ring, the trends in the free energy of association for the formation of 1 : 1 and 1 : 2 lipid-tryptophan adducts both follow an inverted- relationship as a function of the corresponding para-Hammett parameter, with tryptophan (R = H) exhibiting the weakest binding. These trends are shown to be consistent with participation of the indole side chain in both hydrogen bonds and cation-pi interactions. Molecular dynamics simulations of tryptophan and DMPC in an explicit chloroform solvent model demonstrate that for the formation of lipid tryptophan adducts, binding is driven predominantly by carbonyl-cation and cation pi interactions with the choline ammonium group, alongside hydrogen bonding interactions with the lipid phosphate. Some of these interactions operate co operatively, which may account for the observed trends in free energy. PMID- 20024108 TI - Rapid synthesis of Abelson tyrosine kinase inhibitors using click chemistry. AB - Protein kinases catalyze the phosphorylation of serine, threonine, tyrosine and histidine residues in proteins. Aberrant regulation of kinase activity has been implicated in many diseases including cancer. Thus development of new strategies for kinase inhibitor design remains an active area of research with direct relevance to drug development. Abelson (Abl) tyrosine kinase is one of the Src family of tyrosine kinases and is directly implicated in Chronic Myelogenous Leukemia (CML). In this article, we have, for the first time, developed an efficient method for the construction of small molecule-based bisubstrate inhibitors of Abl kinase using click chemistry. Subsequent biochemical screenings revealed a set of moderately potent inhibitors, a few of which have comparable potency to Imatinib (an FDA-approved drug for treatment of chronic myeloid leukemia) against Abl. PMID- 20024109 TI - Sialidase substrate specificity studies using chemoenzymatically synthesized sialosides containing C5-modified sialic acids. AB - para-Nitrophenol-tagged sialyl galactosides containing sialic acid derivatives in which the C5 hydroxyl group of sialic acids was systematically substituted with a hydrogen, a fluorine, a methoxyl or an azido group were successfully synthesized using an efficient chemoenzymatic approach. These compounds were used as valuable probes in high-throughput screening assays to study the importance of the C5 hydroxyl group of sialic acid in the recognition and the cleavage of sialoside substrates by bacterial sialidases. PMID- 20024110 TI - An enantiospecific route towards taiwaniaquinoids. First synthesis of (-) taiwaniaquinone H and (-)-dichroanone. AB - A new methodology for the enantiospecific synthesis of taiwaniaquinoids, based on a thermal 6pi electrocyclization, is reported. Under this procedure, 4a methylhexahydrofluorene terpenoids bearing an A/B trans-configuration has been prepared for the first time. This methodology also makes it feasible to synthesize taiwaniaquinoids with an A/B cis-configuration and 4a methyltetrahydrofluorene terpenoids. Accordingly, the first synthesis of (-) taiwaniaquinone G, (-)-taiwaniaquinone H and (-)-dichroanone has been achieved. PMID- 20024111 TI - Topically resolved intramolecular CH-pi interactions in phenylalanine derivatives. AB - NMR spectra of imines and nitrones derived from benzophenone and phenylalanine or tyrosine show clear evidence of an aromatic edge-to-face interaction in solution. At low temperatures the two ortho protons of the edge interacting phenyl ring become topically resolved with the ortho proton NMR signal involved in the CH-pi interactions shifted well upfield (delta 5.4-5.8 at -88 degrees C) of the other ortho signal. Introduction of a para substituent into the phenylalanine ring has a modest effect on the upfield shift. The edge-to-face arrangement also manifests in the X-ray crystal structures of two of these compounds. Barriers to rotation around the syn phenyl-imino bond are also reported (10.5-11.1 kcal mol(-1)). PMID- 20024112 TI - Towards a chemo-enzymatic method for the asymmetric synthesis of beta-amino tertiary alcohols. AB - The synthesis of a number of beta-amino tertiary alcohols has been achieved via ring-opening of an unsymmetrical epoxide with primary and secondary amines. The results revealed that primary amines give symmetrical triol products following an undesired acyl migration reaction, whereas secondary amines give the desired chiral (racemic) products. Furthermore, we demonstrate that asymmetric products can be formed using enantiomerically pure epoxide and aromatic amines without any loss of enantiomeric excess. PMID- 20024113 TI - Novel dihydro-beta-agarofuran sesquiterpenes as potent modulators of human P glycoprotein dependent multidrug resistance. AB - P-Glycoprotein (Pgp) overexpression is one factor contributing to multidrug resistance (MDR) in cancer cells and represents one drawback in the treatment of cancer. In an attempt to find more specific and less toxic anticancer MDR reversal agents, we report herein the isolation, structure elucidation and biological activity of nine new (, and ) and seven known (, and ) dihydro-beta agarofuran sesquiterpenes from the leaves of Celastrus vulcanicola. Their stereostructures were elucidated on the basis of spectroscopic analysis, including 1D and 2D NMR techniques, CD studies and biogenetic means. All the compounds were assayed on human MDR1-transfected NIH-3T3 cells, in order to determine their ability to reverse the MDR phenotype due to Pgp overexpression. Six compounds from these series (, , , , and ) showed an effectiveness that was similar to (or higher than) the classical Pgp reversal agent verapamil for the reversal of resistance to daunomycin and vinblastine. The structure-activity relationships are discussed. PMID- 20024114 TI - Reactions of 2-(pyrrol-1-yl)benzyl radicals and related species under flash vacuum pyrolysis conditions. AB - 2-(Pyrrol-1-yl)phenoxyl, aminyl, thiophenoxyl and benzyl radicals 2a-2d, respectively, were generated in the gas-phase under flash vacuum pyrolysis conditions. In all cases except the phenoxyl, cyclisation took place providing acceptable synthetic routes to the fused heterocycles 11, 14 and 15, respectively. Only sigmatropic rearrangement products were isolated, in low yields, from the phenoxyl 2a. The pyrrolo[1,2-a]benzimidazole 11 adopts the 1H tautomer exclusively in chloroform solution. Electrophilic substitution reactions of pyrrolo[2,1-b]benzothiophene 14 were studied, including protonation, deuterium exchange, Vilsmeier formylation and reaction with dimethyl acetylenedicarboxylate. 2-(2,5-Diarylpyrrol-1-yl)thiophenoxyl, phenoxyl and aminyl radicals 23a-f, were also generated in the gas-phase under similar conditions. The thiophenoxyls 23a/b gave extremely complex pyrolysate mixtures in which primary cyclisation products were formed by attack of the radical at the pyrrrole ring and attack at the ipso-, ortho- and meta- positions of the aryl ring. Secondary pyrolysis products were obtained by specific sigmatropic shifts of the N-aryl group. The 2,5-di(thien-2-yl)thiophenoxyl radical 23c gave the pyrrolobenzothiazole 31c as the only cyclisation product in low yield. FVP of the phenoxyl and aminyl radical generators 26d and 26f, respectively, gave 3 arylpyrrolo[1,2-f]phenanthridines 46d and 46f, respectively, by a hydrogen transfer-cyclisation mechanism. PMID- 20024115 TI - Sensitive and specific enzyme immunoassays for antigenic trisaccharide from Bacillus anthracis spores. AB - A straightforward synthesis of an anthrose-containing trisaccharide derived from Bacillus anthracis was achieved. Antibodies raised against this hapten provide a highly sensitive enzyme immunoassay with a detection limit of 8.5 pmol mL(-1). By investigating the specificity of the antibodies obtained using different mono-, di- and trisaccharide synthetic analogues, we demonstrated that the epitope was mainly made up of the methyl group at C-5, the butamido group at C-4 and the hydroxyl at C-3 of the anthrose unit, the other parts of the trisaccharide appearing little involved in the recognition. PMID- 20024116 TI - Syntheses of difluorinated carbasugar phosphates from trifluoroethanol. AB - Difluorinated cyclohexene diols (prepared from trifluoroethanol) can be elaborated to racemic analogues of phosphorylated sugars via regioselective protection and phosphorylation of the exposed C-1 hydroxyl group. Cis-diol protection was achieved using stannylene methodology, though the regioselectivity depended on the orientation of the methyl group at C-5. UpJohn dihydroxylation is effective with the phosphotriester in place and global deprotection to the tetrol monophosphates is efficient. PMID- 20024117 TI - A post-modification approach to peptide foldamers. AB - Hydrophilic/hydrophobic patterning is a powerful strategy to control folding in non-natural polymers/oligomers. In this contribution, we present a novel strategy for the preparation of alternating hydrophilic/hydrophobic patterned non-natural peptide foldamers. This strategy relies on the post-modification of a reactive peptide precursor that can be prepared via standard solid phase peptide synthesis without the need for side chain protective groups. The peptide scaffolds consisted of an alternating sequence of l-leucine and l-allylglycine residues. Using thiol-ene chemistry, the double bonds in the side chains of the l allylglycine units could be post-modified with cysteamine hydrochloride, thioglycolic acid, 1-thioglycerol or 2,3,4,6-tetra-O-acetyl-thio-beta-d glucopyranose to afford alternating hydrophilic/hydrophobic patterned peptides. In agreement with the alternating hydrophilic/hydrophobic patterned primary structure, cysteamine and thioglycolic acid post-modified octapeptides were found to adopt a beta-sheet secondary structure in basic or acidic aqueous media, respectively. The proposed synthetic approach is not only of interest to generate diverse libraries of peptide foldamers from a limited number of reactive precursor scaffolds, but may also represent an attractive, general strategy for the synthesis of peptides with complex side chain functionalities that are not easily accessible via standard solid phase techniques. PMID- 20024118 TI - Synthesis and evaluation of fused bispyrimidinoacridines as novel pentacyclic analogues of quadruplex-binder BRACO-19. AB - The present article reports on the design and the synthesis of a series of mono- and bis-pyrimidinoacridines and their evaluation as a novel family of quadruplex binders. It is shown that bispyrimidinoacridines represent an interesting compromise between easy synthetic access and efficiency in terms of quadruplex interaction (both affinic and selective), as judged by G4-FID assay and molecular modelling. The present study also highlights that control of the pi-stacking interactions taking place between the ligand and the accessible G-tetrad of a quadruplex-DNA is indeed essential for good recognition but not exclusively (key role of direct and water-mediated H-bonds). The introduction of additional amino side chains, valuable in the acridine series, results here in steric perturbations of the ligand/quadruplex recognition and lowers the quadruplex/duplex selectivity. PMID- 20024119 TI - Macrocyclic aromatic polysulfones and sulfide-sulfones: synthesis and structural characterisation of molecular pentagons and rectangles. AB - Cyclo-condensation of arylenedithiols with bis(4-chlorophenylenesulfone)s under pseudo-high-dilution conditions affords macrocyclic aromatic sulfide-sulfones which are readily oxidised to all-sulfone-linked macrocycles. The cyclic pentamer of poly(1,4-phenylenesulfone) and cyclic dimer of poly(1,4-phenylenesulfonyl-4,4' biphenylenesulfone) have been isolated and characterised. PMID- 20024120 TI - Mechanism of glycoside hydrolysis: A comparative QM/MM molecular dynamics analysis for wild type and Y69F mutant retaining xylanases. AB - Computational simulations have been performed using hybrid quantum mechanical/molecular-mechanical potentials to investigate the catalytic mechanism of the retaining endo-beta-1, 4-xylanase (BCX) from B. circulans. Two-dimensional potential-of-mean-force calculations based upon molecular dynamics with the AM1/OPLS method for wild-type BCX with a p-nitrophenyl xylobioside substrate in water clearly indicates a stepwise mechanism for glycosylation: the rate determining step is nucleophilic substitution by Glu78 to form the covalently bonded enzyme-substrate intermediate without protonation of the leaving group by Glu172. The geometrical configuration of the transition state for the enzymic reaction is essentially the same as found for a gas-phase model involving only the substrate and a propionate/propionic acid pair to represent the catalytic glutamate/glutamic acid groups. In addition to stabilizing the (2,5)B boat conformation of the proximal xylose in the non-covalent reactant complex of the substrate with BCX, Tyr69 lowers the free-energy barrier for glycosylation by 42 kJ mol(-1) relative to that calculated for the Y69F mutant, which lacks the oxygen atom O(Y). B3LYP/6-31+G* energy corrections reduce the absolute height of the barrier to reaction. In the oxacarbenium ion-like transition state O(Y) approaches closer to the endocyclic oxygen O(ring) of the sugar ring but donates its hydrogen bond not to O(ring) but rather to the nucleophilic oxygen of Glu78. Comparison of the average atomic charge distributions for the wild-type and mutant indicates that charge separation along the bond between the anomeric carbon and O(ring) is matched in the former by a complementary separation of charge along the O(Y)-H(Y) bond, corresponding to a pair of roughly antiparallel bond dipoles, which is not present in the latter. PMID- 20024121 TI - Assessing the cluster glycoside effect during the binding of concanavalin A to mannosylated artificial lipid rafts. AB - Mannosyl glycolipids with perfluoroalkyl membrane anchors have been synthesised. When inserted into vesicles, these mannosyl lipids either dispersed evenly over the surface or, in the presence of cholesterol, phase-separated into artificial lipid rafts. At 1% mol/mol, the affinity of dispersed mannosyl lipids for Con A was 3-fold weaker than in solution, perhaps reflecting steric blocking by the surface. However increasing membrane loading 5-fold increased Con A affinity by up to 75% and indicated weak intramembrane chelation of Con A. Despite this observation, concentrating the mannosyl lipids into artificial lipid rafts did not significantly improve affinity for Con A. This lack of a cluster glycoside effect was ascribed to lipid congestion inhibiting intra-raft chelation of Con A, and implies that glycolipids located in lipid rafts may not necessarily be preorganised for multivalent binding. PMID- 20024122 TI - Cyclooctyne-based reagents for uncatalyzed click chemistry: A computational survey. AB - With the goal of identifying alkyne-like reagents for use in click chemistry, but without Cu catalysts, we used B3LYP density function theory (DFT) to investigate the trends in activation barriers for the 1,3-dipolar cycloadditions of azides with various cyclooctyne, dibenzocyclooctyne, and azacyclooctyne compounds. Based on these trends, we find monobenzocyclooctyne-based reagents that are predicted to have dramatically improved reactivity over currently employed reagents. PMID- 20024123 TI - Synthesis and characterization of coumarin-based europium complexes and luminescence measurements in aqueous media. AB - A series of new ligands suitable for the formation of luminescent lanthanide complexes in water is described. The chelates are designed for analyte labeling and play the role of fluorescent donor in homogeneous time-resolved fluorescence assays using LEDs as a light source for excitation at 370 nm. Ligands are constructed from a coumarin nucleus, for lanthanide sensitization, and different aminomethylenecarboxy moieties are introduced in positions 7 and 5, 6, or 8 of the sensitizer. A reactive spacer arm under biocompatible conditions (maleimide, azide) is introduced at position 3 for ultimate bioconjugation purposes. The synthesis and characterization of the ligands are described, together with the preparation of their corresponding europium complexes. Photophysical properties of the complexes are investigated in water by means of UV-vis and luminescence spectroscopy. PMID- 20024125 TI - Synthetic biology. AB - There is currently much excitement surrounding the rapidly growing discipline of synthetic biology, which utilizes the design and construction principles of engineering to develop, evolve and standardize biological components and systems. This systematic approach to improving and increasing the programmability and robustness of biological components is expected to lead to the facile assembly of artificial biological components and integrated systems that enable innovative approaches to solving a wide range of societal challenges. Here we discuss the current state of the art and outline the next wave of synthetic biology: integrating individual components into systems. PMID- 20024126 TI - Synthesis of natural products containing spiroketals via intramolecular hydrogen abstraction. AB - Although known for over a quarter of a century, the oxidative radical cyclisation route to spiroketals has found limited use in natural product synthesis in comparison to classical approaches. Its successful application in this field of research forms the subject of this perspective. PMID- 20024127 TI - Highly efficient synthesis of medium-sized lactones via oxidative lactonization: concise total synthesis of isolaurepan. AB - A catalytic amount of TEMPO in the presence of PhI(OAc)(2) effected oxidative lactonization of 1,6- and 1,7-diols, directly affording seven- and eight-membered lactones, respectively, in good yields. PMID- 20024128 TI - Impregnated copper on magnetite: an efficient and green catalyst for the multicomponent preparation of propargylamines under solvent free conditions. AB - Impregnated copper on magnetite is a versatile, inexpensive and simple catalyst for the selective multicomponent reaction of terminal alkynes, aldehydes and secondary amines to give the corresponding propargylamines with excellent yields. The catalyst can be easily recovered and reused by using a simple magnet. The process could be repeated up to ten times without losing its activity. PMID- 20024129 TI - Catalytic asymmetric conjugate addition of dialkylzinc reagents to alpha,beta unsaturated sulfones. AB - An efficient method is reported for the highly enantioselective copper-catalyzed conjugate addition of dialkylzinc reagents to alpha,beta-unsaturated sulfones using a monodentate phosphoramidite ligand. PMID- 20024130 TI - Small molecule induced control in duplex and triplex DNA-directed chemical reactions. AB - Triplex DNA binders can effectively control copper-catalysed alkyne-azide click reactions in DNA architecture, such that either duplex or triplex DNA directed reactions of terminally attached azides and alkynes occur, in the absence or presence of triplex DNA binder, respectively. PMID- 20024131 TI - Synthesis, structure and properties of decakis(phenylthio)corannulene. AB - Decakis(phenylthio)corannulene has been prepared from decachlorocorannulene by direct nucleophilic substitution; electronic structure properties and the X-ray crystal structure were determined and compared to predictions made by ab initio quantum chemical calculations. PMID- 20024132 TI - Synthesis and application of a new cleavable linker for "click"-based affinity chromatography. AB - A new chemically-cleavable linker has been synthesised for the affinity independent elution of biomolecules by classical affinity chromatography. This azo-based linker is shown to couple efficiently with "click" derivatised ligands such as biotin propargyl amide through a copper(I)-catalysed Huisgen 1,3-dipolar cycloaddition reaction. Binding to Affi-Gel matrices displaying ligands coupled to the new linker is both efficient and selective. The captured material may be readily released from the resin upon treatment with sodium dithionite. These mild elution conditions have allowed for the efficient isolation of the affinity partner from complex protein mixtures such as those found in fetal bovine serum. PMID- 20024133 TI - Development of a new microwave-assisted cleavable backbone amide linker (BAL): a comparative study. AB - A thorough comparative study to demonstrate the properties of a new microwave labile backbone amide linker is presented. A cyclic pentapeptide, cyclo(Trp-Gln Gly-beta-Ala-Phe), was used as a model and synthesized following 16 different conditions. The new backbone amide linker is stable towards acid and base at r.t., and can be cleaved at elevated temperature in trifluoroacetic acid under microwave irradiation, avoiding the use of aggressive reagents like HF. The new linker is compatible with Boc- as well as Fmoc-strategy and allows the cleavage of acid labile side chain protective groups at r.t., prior to cleavage of the cyclic pentapeptide from the resin. PMID- 20024134 TI - Synthesis and biological evaluation of phosphatidylinositol phosphate affinity probes. AB - The synthesis of the complete family of phosphatidylinositol phosphate analogues (PIPs) from five key core intermediates A-E is described. These core compounds were obtained from myo-inositol orthoformate 1 via regioselective DIBAL-H and trimethylaluminium-mediated cleavages and a resolution-protection process using camphor acetals 10. Coupling of cores A-E with phosphoramidites 34 and 38, derived from the requisite protected lipid side chains, afforded the fully protected PIPs. Removal of the remaining protecting groups was achieved via hydrogenolysis using palladium black or palladium hydroxide on carbon in the presence of sodium bicarbonate to afford the complete family of dipalmitoyl- and amino-PIP analogues 42, 45, 50, 51, 58, 59, 67, 68, 76, 77, 82, 83, 92, 93, 99 and 100. Investigations using affinity probes incorporating these compounds have identified novel proteins involved in the PI3K intracellular signalling network and have allowed a comprehensive proteomic analysis of phosphoinositide interacting proteins. PMID- 20024135 TI - Asymmetric Michael addition reaction of 3-substituted-N-Boc oxindoles to activated terminal alkenes catalyzed by a bifunctional tertiary-amine thiourea catalyst. AB - The current article reports an organocatalytic strategy for the asymmetric catalysis of chiral oxindoles bearing 3-position all-carbon quaternary stereocenters. Accordingly, highly enantioselective Michael addition reactions of 3-substituted oxindoles to terminal alkenes have been developed by utilizing a bifunctional tertiary-amine thiourea catalyst. The reactions accommodate a number of Michael donor compounds (different substituted 3-aryl or methyl oxindoles), and Michael acceptor compounds (vinyl ketones and vinyl sulfones) to give the desired oxindole products with moderate to excellent yields (up to 99%) and moderate to excellent enantioselectivities (up to 91% ee). PMID- 20024136 TI - The stability of imine-containing dynamic [2]rotaxanes to hydrolysis. AB - Large amounts (>100 mol equivalents) of water are required to effect by hydrolysis the partial disassembly of the rings from the dumbbell components of two dynamic [2]rotaxanes. The two dynamic [2]rotaxanes are comprised of [24]crown 8 rings-each of which incorporate two imine bonds-encircling a dumbbell component composed of a dibenzylammonium ion in which each of the two benzyl substituents carries two methoxyl groups attached to their 3- and 5-positions. A mechanism for the partial disassembly of the two dynamic [2]rotaxanes, involving the cleavage of the kinetically labile imine bonds by water molecules, is proposed. The most important experimental observation to be noted is the fact that the hydrolysis of the macrocyclic diimines, associated with the templating -CH(2)NH(2)(+)CH(2) centres in the middle of their dumbbells, turns out to be an uphill task to perform in the face of the molecular recognition provided by strong [N(+)-HO] hydrogen bonds and weaker, yet not insignificant, [C-HO] interactions. The dynamic nature of the imine bond formation and hydrolysis is such that the acyclic components produced during hydrolysis of the imine bonds can be enticed to cyclise once again around the -CH(2)NH(2)(+)CH(2)-template, affording the [2]rotaxanes. The reluctance of imine bonds, present in substantial numbers in larger molecular and extended structures, is significant when it comes to exercising dynamic chemistry in compounds where multiple imine bonds are present. PMID- 20024137 TI - Microwave-assisted three-component coupling-addition-S(N)Ar (CASNAR) sequences to annelated 4H-thiopyran-4-ones. AB - A whole family of annelated 4H-thiopyran-4-ones as the core structural unit was readily synthesized in good yields by a microwave-assisted coupling-addition S(N)Ar (CASNAR) sequence starting from readily available (het)aroyl chlorides, alkynes and sodium sulfide nonahydrate in a consecutive one-pot three-component reaction. All representatives display a pronounced halochromicity of the absorption bands upon protonation. According to DFT calculations, the electronic ground state of the annelated 4H-thiopyran-4-ones possess a considerable zwitterionic character. PMID- 20024138 TI - Bis-cation salt complexation by meso-octamethylcalix[4]pyrrole: linking complexes in solution and in the solid state. AB - Pyridinium and imidazolium bis-cations are shown to link calix[4]pyrrole anion complexes both in solution and in the solid state. This is accomplished by binding of the bis-cations to the electron-rich bowl shaped cavities formed by two separate calixpyrrole-anion complexes. These resulting sandwich-type structures provide a new way of organising calix[4]pyrrole anion complexes in space. PMID- 20024139 TI - Carbamate complexation by urea-based receptors: studies in solution and the solid state. AB - The interactions of a series of urea based neutral hydrogen bond donor anion receptors have been investigated with i) alkylcarbamate anions formed by the reaction of carbon dioxide with primary aliphatic amines and ii) the zwitterionic species formed by the reaction of carbon dioxide with 1,4,5,6 tetrahydropyrimidine. Significant downfield chemical shift changes were observed for the urea NH protons in many cases, consistent with host : anion hydrogen bonding interactions, and thus stabilisation of the carbon dioxide bound species. In the case of the alkylammonium-alkylcarbamate salts, this represents successful competition with electrostatic interactions between the alkylcarbamate and alkylammonium components of the salt. A synchrotron structure of a ternary complex formed by an amide appended diindolylurea, the ammonium carbamate salt formed by 1,3-diaminopropane and CO(2) and 18-crown-6, was elucidated and shows the carbamate group bound by six hydrogen bonds (accepting five and donating one) to the functionalised diindolylurea. PMID- 20024140 TI - Synthesis of dihydrodehydrodiconiferyl alcohol: the revised structure of lawsonicin. AB - Structural revision of lawsonicin, a natural product of Lawsonia alba, is reported based upon comparison of its spectral data with that of the naturally occurring dihydrobenzo[b]furan neolignan (rac)-trans-dihydrodehydrodiconiferyl alcohol, which is found to be identical. A concise synthesis of dihydrodehydrodiconiferyl alcohol, via Rh(2)[S-DOSP](4)-catalysed intramolecular C-H insertion, is described. PMID- 20024141 TI - On the importance of simultaneous infrared/fiber-optic temperature monitoring in the microwave-assisted synthesis of ionic liquids. AB - The temperature profiles obtained from both an external infrared and internal fiber-optic sensor were compared for heating and synthesizing the ionic liquid 1 butyl-3-methylimidazolium bromide (bmimBr) under microwave conditions. Utilizing a single-mode microwave reactor that allows simultaneous infrared/fiber-optic temperature measurements, significant differences between the two methods of temperature monitoring were revealed. Due to the strong microwave absorptivity of ionic liquids and the delay experienced in monitoring temperature on the outer surface of a heavy-walled glass vial, external infrared temperature sensors can not be used to accurately control the temperature in the heating of ionic liquids under microwave conditions. The use of internal fiber-optic probes allows the monitoring and control of the heating behavior in a much better way. In order to prevent the strong exotherm in the synthesis of bmimBr under microwave conditions the use of a reaction vessel made out of silicon carbide is the method of choice. Because of the high thermal conductivity and effusivity of silicon carbide, the heat generated during the ionic liquid formation is efficiently exchanged with the comparatively cool air in the microwave cavity via the silicon carbide ceramic. PMID- 20024142 TI - The crystal structure of an LLL-configured depsipeptide substrate analogue bound to isopenicillin N synthase. AB - Isopenicillin N synthase (IPNS) is a non-heme iron(ii) oxidase, which catalyses the biosynthesis of isopenicillin N (IPN) from the tripeptide delta-l-alpha aminoadipoyl-l-cysteinyl-d-valine (lld-ACV) in a remarkable oxidative bicyclisation reaction. The natural substrate for IPNS is the lld-configured tripeptide. lll-ACV is not turned over by the enzyme, but inhibits turnover of the lld-tripeptide. The mechanism by which this inhibition takes place is not fully understood. Recent studies have employed a range of lld-configured depsipeptide substrate analogues in crystallographic studies to probe events preceding beta-lactam closure in the IPNS reaction cycle. Herein, we report the first crystal structure of IPNS in complex with an lll-configured depsipeptide analogue, delta-l-alpha-aminoadipoyl-l-cysteine (1-(R)-carboxy-2-thiomethyl)ethyl ester (lll-ACOmC). This report describes the crystal structure of the IPNS:Fe(ii):lll-ACOmC complex to 2.0 A resolution, and discusses attempts to oxygenate this complex at high pressure in order to probe the mechanism by which lll-configured substrates inhibit IPNS catalysis. PMID- 20024143 TI - Total synthesis of Le(A)-LacNAc pentasaccharide as a ligand for Clostridium difficile toxin A. AB - The toxins TcdA and TcdB produced by the human pathogen Clostridium difficile gain entrance to host epithelial cells by recognizing cell-surface carbohydrate ligands. Inhibiting the attachment of these toxins to host cells has been proposed to be a viable therapy to treat C. difficile infections. Glycan array screening previously revealed that the Le(A)-LacNAc pentasaccharide binds strongly to TcdA. Here we report the efficient syntheses of the pentasaccharide and a structurally related tetrasaccharide motif. These compounds will be used to better define the carbohydrate-binding specificity of toxins from C. difficile, which will hopefully lead to the development of improved therapeutics. PMID- 20024144 TI - Dendron-anchored organocatalysts: the asymmetric reduction of imines with trichlorosilane, catalysed by an amino acid-derived formamide appended to a dendron. AB - Asymmetric reduction of ketimines with trichlorosilane can be catalysed by the Lewis-basic N-methylvaline-derived formamide anchored to a soluble dendron () with good enantioselectivity (S equilibrium of the fluorinated sugar ring. PMID- 20024148 TI - DIBAL-H mediated triple and quadruple debenzylations of perbenzylated cyclodextrins. AB - Diisobutylaluminium hydride (DIBAL-H) mediated reductive removal of benzyl groups was investigated for perbenzylated alpha-, beta- and gamma-cyclodextrins using DIBAL-H in hexane as the reagent. It was found that under the new conditions, the debenzylation can be better controlled to provide sequentially tri- and tetra debenzylated products in moderate yields and in a regioselective manner. In the case of alpha-cyclodextrin, the removal of the third and fourth benzyl groups took a different path involving the secondary rim, compared to beta- and gamma cyclodextrins which both gave only 6-O-debenzylated products. PMID- 20024149 TI - Epimeric and amino disaccharide analogs as probes of an alpha-(1-->6) mannosyltransferase involved in mycobacterial lipoarabinomannan biosynthesis. AB - Mycobacterial lipoarabinomannan (LAM) is an important, immunologically active glycan found in the cell wall of mycobacteria, including the human pathogen Mycobacterium tuberculosis. At the core of LAM is a mannan domain comprised of alpha-(1-->6)-linked-mannopyranose (Manp) residues. Previously, we and others have demonstrated that alpha-Manp-(1-->6)-alpha-Manp disaccharides (e.g., Manp-(1 ->6)-alpha-ManpOctyl, ) are the minimum acceptor substrates for enzymes involved in the assembly of the LAM mannan core. We report here the synthesis five epimeric and three amino analogs of , and their subsequent biochemical evaluation against an alpha-(1-->6)-ManT activity present in a membrane preparation from M. smegmatis. Changing the manno- configuration of either residue of to talo- or gluco- led to a reduction or loss of activity, thus confirming earlier work showing that the C-2 and C-4 hydroxyl groups of each monosaccharide were important for enzymatic recognition. Characterization of the products formed from these analogs was done using a combination of mass spectrometry and glycosidase digestion, and full substrate kinetics were also performed. The analogs in which the acceptor hydroxyl group had been replaced with an amino group were, as expected, not substrates for the enzyme, but were weak inhibitors. PMID- 20024150 TI - Asymmetric ruthenium-catalyzed 1,4-additions of aryl thiols to enones. AB - Well defined, stable, one-point binding ruthenium complexes and selectively bind and activate alpha,beta-unsaturated carbonyl compounds for cycloaddition reactions. These mild Lewis acids catalyze asymmetric 1,4-addition reactions of aryl thiols to enones with product selectivities up to 87% ee. (31)P NMR experiments provide an insight into the intricate equilibria governing the reaction mechanism. The absolute configuration of the major products indicates enones to react in the syn-s-trans orientation. Models based on X-ray structures of the Ru complexes can be used to rationalize selectivity. PMID- 20024151 TI - A novel bimodal lipidic contrast agent for cellular labelling and tumour MRI. AB - We have synthesized a bimodal lipidic molecule bearing both fluorophore and contrast agent signatures on the same structure in order to create a robust bimodal liposome for both magnetic resonance imaging (MRI) and fluorescence microscopy utility. The dual-modality concept considered in the synthesis of this new paramagnetic and fluorescent lipid is valuable in that anatomical information (MRI) as well as very sensitive localization (ex vivo fluorescence microscopy) of signal, and therefore liposome biodistribution, is obtainable. Bimodal cationic and neutral PEGylated liposomes were formulated using this novel lipid probe and used to label cells in vitro and image human ovarian xenografts in vivo. Tumour signal enhancement was increased by over 6-fold post-administration of the neutral PEGylated liposomes, and was maintained at this level up to the 24 h end point. Our results showed this lipid to be more effective and sensitive than the single signature paramagnetic lipid Gd.DOTA.DSA at cellular labelling and tumour MRI. PMID- 20024152 TI - Stereoselective synthesis and structure-affinity relationships of bicyclic kappa receptor agonists. AB - Reductive amination of the bicyclic ketone 4 led diastereoselectively to endo configured amines, which were transformed into the amides 7-10. The synthesis of the diastereomers 25 with an exo-configured amino moiety at position 6 was only successful after deactivation of both N-atoms of the 1,4 diazabicyclo[3.3.1]nonane system. The N-1-oxide 19 with an N-4-tosyl moiety was the crucial intermediate, which allows SN2 substitution with NaN3 under inversion of the configuration at position 6. Whereas the endo-configured pyrrolidine 7a (WMS-1302) revealed a kappa receptor affinity of 73 nM, the exo-configured diastereomer 25a was almost inactive at the kappa receptor (Ki > 1 microM). Replacement of the 3,4-dichlorophenylacetyl residue by other acyl and sulfonyl residues showed that it is essential for high kappa affinity. The kappa receptor affinities of the conformationally constrained pyrrolidines 7a and 25a were correlated with the dihedral angle N(pyrrolidine)-C-C-N(acetamide). A systematic conformational analysis of the potent but flexible kappa agonist 2 showed that a dihedral angle of 168 degrees (as in 25a) is energetically more disfavored than a dihedral angle of 58 degrees (7a). However, even the conformation with a dihedral angle of 58 degrees does not represent an energy minimum, which might explain the reduced kappa affinity of 7a. PMID- 20024153 TI - Oxidative spirocyclisation routes towards the sawaranospirolides. Synthesis of ent-sawaranospirolides C and D. AB - Two routes are described for the synthesis of the sawaranospirolides, stereoisomeric spirolactone ascorbigenins isolated from Chamaecyparis pisifera. Trapping of the keto enal formed by oxidation of a functionalised 2-(4 hydroxybutyl)furan affords a potential butenolide spiroacetal precursor to sawaranospirolides A and C. Alternatively, epoxidation of protected 3 (dihydropyran-2-yl)-3-arylpropanoic acids results in spirolactonisation to generate ent-sawaranospirolide C; a related acid-mediated spirocyclisation gave access to ent-sawaranospirolide D. PMID- 20024154 TI - Identification and biosynthesis of tropone derivatives and sulfur volatiles produced by bacteria of the marine Roseobacter clade. AB - Bacteria of the Roseobacter clade are abundant marine bacteria and are important contributors to the global sulfur cycle. The volatiles produced by two of its members, Phaeobacter gallaeciensis and Oceanibulbus indolifex, were analyzed to investigate whether the released compounds are derived from sulfur metabolism, and which biosynthetic pathways are involved in their formation. Both bacteria emitted different sulfides and thioesters, including new natural compounds such as S-methyl phenylethanethioate (16) and butyl methanesulfonate (21). The S methyl alkanoates were identified by comparison with standards that were synthesized from the respective methyl alkanoates by a new method using an easily prepared aluminium/sulfur reagent. Phaeobacter gallaeciensis is also able to produce tropone (37) in large amounts. Its biosynthesis was investigated by various feeding experiments, showing that 37 is formed via a deviation of the phenylacetate catabolism. The unstable tropone hydrate 42 was identified as an intermediate of the tropone biosynthesis that was also released together with tropolone (38). PMID- 20024155 TI - Host-guest complexations of local anaesthetics by cucurbit[7]uril in aqueous solution. AB - The cucurbit[7]uril (CB[7]) host molecule forms very stable host-guest complexes with the local anaesthetics procaine (K(CB[7]) = (3.5 +/- 0.7) x 10(4) dm(3) mol( 1)), tetracaine (K(CB[7]) = (1.5 +/- 0.4) x 10(4) dm(3) mol(-1)), procainamide (K(CB[7]) = (7.8 +/- 1.6) x 10(4) dm(3) mol(-1)), dibucaine (K(CB[7]) = (1.8 +/- 0.4) x 10(5) dm(3) mol(-1)) and prilocaine (K(CB[7]) = (2.6 +/- 0.6) x 10(4) dm(3) mol(-1)) in aqueous solution (pD = 4.75). The stability constants are 2-3 orders of magnitude greater than the values reported for binding by the comparably sized beta-cyclodextrin host molecule. The inclusion by CB[7] raises the first pK(a) values of the anaesthetics by 0.5-1.9 pK units, as the protonated forms are bound more strongly in acidic solution. The complexation-induced chemical shift changes in the guest proton resonances provide an indication of the site(s) of binding and the effects of protonation on the location of the binding sites. PMID- 20024156 TI - Cucurbit[7]uril host-guest complexes of cholines and phosphonium cholines in aqueous solution. AB - The neutral host cucurbit[7]uril forms very stable complexes with a series of cationic cholines (R(3)NCH(2)CH(2)OR'(+)) and their phosphonium analogues (R(3)PCH(2)CH(2)OR'(+)) (R(3) = Me(3), Et(3), or Me(2)Bz, or R(3)N = quinuclidinium, and R' = H, COCH(3), CO(CH(2))(2)CH(3), or PO(3)H), and (+/-) carnitine, in aqueous solution. The complexation behaviour has been investigated using (1)H and (31)P NMR spectroscopies, and ESI mass spectrometry. The complexation-induced chemical shift changes of the guests clearly indicate the effects of replacing the N(CH(3))(3)(+) end group by P(CH(3))(3)(+), and changing the nature of R on the position of the guest with respect to the CB[7] cavity and its polar portal-lining carbonyl groups. This study demonstrates that molecular recognition of cholines in aqueous solution is achievable with a neutral host without the need for aromatic walls for cation-pi interactions. PMID- 20024157 TI - Prediction of the main macrocyclic conjugation pathway for porphyrinoids from the ring current distribution. AB - For some porphyrinoids, such as orangarin and amethyrin, the main route of macrocyclic pi-circulation is different from the main macrocyclic conjugation pathway predicted by porphyrin chemists. Our analytical theory of ring-current diamagnetism allows us to predict the main macrocyclic conjugation pathway from the ring current distribution. We can now interpret macrocyclic aromaticity and macrocyclic circulation consistently within the same theoretical framework. PMID- 20024158 TI - Facile P,N-heterocycle synthesis via tandem aminomethylation-cyclization of H phosphinate building blocks. AB - Various heterocycles containing phosphorus and nitrogen are easily synthesized from readily available H-phosphinate building blocks. Aminomethylation of these H phosphinates is followed by in situ cyclization through substitution or cross coupling to produce novel heterocycles in moderate to good yields. PMID- 20024159 TI - Study on the selectivity in the electrophilic monofluorination of 2,3-allenoates with Selectfluor: an efficient synthesis of 4-fluoro-2(5H)-furanones and 3-fluoro 4-oxo-2(E)-alkenoates. AB - Different from the reaction of 2,3-allenoic acids with Selectfluor, 4-fluoro 2(5H)-furanones and (E)-3-fluoro-4-oxo-2-alkenoates were highly selectively generated from 2,4-disubstituted 2,3-allenoates with Selectfluor under different conditions in moderate yields. The reaction of 2,4,4-trisubstituted 2,3 allenoates afforded the corresponding 4-fluoro-2(5H)-furanones highly selectively with up to 95% yield under different conditions. The scope of the substrates has been carefully explored. Due to the more readily availability of 2,3-allenoates as compared to 2,3-allenoic acids, new 4-fluoro-2(5H)-furanones were prepared. Based on the isolation and characterization of the minor fluorohydroxylation product E-5m, a mechanism has been proposed. PMID- 20024160 TI - A novel biomimetic photochemical switch at work: design of a photomodulable peptide. AB - We report the outcomes of our recent computational and experimental work for the development of a novel biomimetic molecular switch. Furthermore, we present the new results on the design and computational characterization of a "functional" cyclic peptidomimetic formed by the switch conjugated to a biologically active peptide: the RGD sequence involved in the control of cell adhesion. Structural properties of the construct are investigated in aqueous solution using molecular dynamics (MD) simulations. Analysis of MD trajectories reveals that, for each diastereoisomer of the switch (E or Z), different conformations are stabilized. Electrostatic and spectroscopic properties of such conformers are evaluated by means of ab initio multiconfiguration quantum chemical method implemented in a quantum-mechanical/molecular-mechanic (CASPT2//CASSCF/6-31G*/AMBER) scheme. PMID- 20024161 TI - Enhanced efficiency of the visible-light photocatalytic hydrogen generation by the ruthenium tris(2,2'-bipyridyl)-methyl viologen system in the presence of cucurbit[n]urils. AB - Cucurbiturils are cyclic oligomers of glycoluril units whose molecular shape defines an internal hollow space accessible through carbonyl portals. It is known that methyl viologen forms strong host-guest complexes with CB[7] and CB[8]. In the latter case even a dimer can be hosted inside CB[8]. Classical solution systems for visible light H(2) generation use methyl viologen as an electron relay. In this work we have found that the efficiency for H(2) generation of the photocatalytic system comprising EDTA-ruthenium tris(2,2'-bipyridyl)-methyl viologen-Pt increases when cucurbiturils are present in the solution. The enhancement follows the order CB[6] < CB[7] < CB[8]. By means of laser flash photolysis we have established that this efficiency increase arises from the combination of an increase in the relative quantum yield of charge separation (higher efficiency in the formation of MV*(+)) and a decrease of MV*(+) lifetime. PMID- 20024162 TI - The photophysics of alloxazine: a quantum chemical investigation in vacuum and solution. AB - (Time-dependent) Kohn-Sham density functional theory and a combined density functional/multi-reference configuration interaction method (DFT/MRCI) were employed to explore the ground and low-lying electronically excited states of alloxazine, a flavin related molecule. Spin-orbit coupling was taken into account using an efficient, nonempirical mean-field Hamiltonian. Intersystem crossing (ISC) rate constants for S --> T transitions were computed, employing both direct and vibronic spin-orbit coupling. Solvent effects were mimicked by a conductor like screening model and micro-hydration with up to six explicit water molecules. Multiple minima were found on the first excited singlet (S(1)) potential energy hypersurface (PEH) with electronic structures (1)(npi*) and (1)(pipi*), corresponding to the dark 1 (1)A'' (S(1)) state and the nearly degenerate, optically bright 2 (1)A' (S(2)) state in the vertical absorption spectrum, respectively. In the vacuum the minimum of the (1)(npi*) electronic structure is clearly found below that of the (1)(pipi*) electronic structure. Population transfer from (1)(pipi*) to (1)(npi*) may proceed along an almost barrierless pathway. Hence, in the vacuum, internal conversion (IC) between the 2 (1)A' and the 1 (1)A'' state is expected to be ultrafast and fluorescence should be quenched completely. The depletion of the (1)(npi*) state is anticipated to occur via competing IC and direct ISC processes. In aqueous solution this changes, due to the blue shift of the (1)(npi*) state and the red shift of the (1)(pipi*) state. However, the minimum of the (1)(npi*) state still is expected to be found on the S(1) PEH. For vibrationally relaxed alloxazines pronounced fluorescence and ISC by a vibronic spin-orbit coupling mechanism is expected. At elevated temperatures or excess energy of the excitation laser, the (1)(npi*) state is anticipated to participate in the deactivation process and to partially quench the fluorescence. PMID- 20024163 TI - Kinetics of reversible photoisomerization: determination of the primary quantum yields for the E-Z photoisomerization of silylenephenylenevinylene derivatives. AB - The kinetics scheme for directly excited, photoreversible reactions is solved exactly under the assumptions of no irreversible side reactions and constant excitation intensity for the duration of the reaction. The advantages of the methodology over the extrapolation-to-zero-time and the back-reaction correction methods are (i) that the quantum yields of both the forward and reverse photoreactions can be obtained starting from either pure reactant or pure product and (ii) the conversion percentage is not limited to a narrow domain in the neighborhood of small conversions. Examples of E-Z photoisomerizations are given to illustrate the fitting procedures required. The results from these examples are compared to the photoisomerization method of extrapolating the empirical quantum yields to zero time and the back-reaction correction. The exact equations are used to justify the extrapolation-to-zero-time method and to establish criteria on extrapolation ranges for the conversion percentage of starting material. PMID- 20024164 TI - A novel elsinochrome A derivative: a study of drug delivery and photodynamic activity. AB - Elsinochrome A (EA) possesses the highest singlet-oxygen quantum yield (0.98) amongst the perilenoquinoid pigments and may be suitable as a phototherapeutic drug. However, there have been virtually no studies into its medicinal applications. Based on the analysis of chemical derivatives of hypocrellins (the same family as EA), 5-(3-mercapto-1-propanesulfonic acid)-substituted elsinochrome A (MPEA) with an amphiphilicity was designed and synthesized by considering drug delivery and biological activity requirements. MPEA possesses a water solubility of 5.1 mg mL(-1), which is just sufficient to enable dissolution at a clinically acceptable concentration, while its partition coefficient (n octanol/phosphate buffered saline) of 7 guarantees affinity to biological targets. MPEA could photogenerate semiquinone anion radicals and reactive oxygen species, especially singlet oxygen, at a yield of 0.73, which approaches that for hypocrellin B. Biological tests confirmed that the photodynamic activity of MPEA was as high as 60% of that of its parent EA, which is significantly higher than that of most other photosensitizers. PMID- 20024166 TI - Effect of bridgehead substitution on the fluorescence quenching of 2,3 diazabicyclo[2.2.2]oct-2-enes by solvents and antioxidants. AB - Azoalkanes of the 2,3-diazabicyclo[2.2.2]-oct-2-ene type have been introduced as probes for antioxidants in homogeneous solution as well as in liposomes and micelles. The bimolecular fluorescence quenching of the bridgehead dichloro substituted 1,4-dichloro-2,3-diazabicyclo[2.2.2]-oct-2-ene (3) was compared with that of the parent compound 2,3-diazabicyclo[2.2.2]-oct-2-ene (1) and the bridgehead-dialkylated compound 4-methyl-1-isopropyl-2,3-diazabicyclo[2.2.2]-oct 2-ene (2). Compound 3 showed a more efficient fluorescence quenching in C-H containing solvents (e.g., in n-hexane: 30 ns for 3 versus 340 ns for 1 and 770 ns for 2), but a less efficient quenching in aqueous solution (e.g., in deaerated H(2)O: 485 ns for 3 versus 420 ns for 1 and 340 ns for 2), and also by molecular oxygen (k(q)/10(9) M(-1) s(-1) = 0.32 for versus 2.5 for 1 and 1.9 for 2). Towards low-molecular weight antioxidants, compound 3 showed a significantly higher reactivity (e.g., for reduced glutathione: k(q)/10(9) M(-1) s(-1) = 1.8 for 3 versus 0.82 for 1 and 0.39 for 2), at the expense of a lower differentiation between the investigated antioxidants (lower selectivity). The increased reactivity of 3 and lower, as well as qualitatively different, selectivity is attributed to a combination of factors, most importantly the slightly increased excitation energy of 3 and its lower excited-state nucleophilicity. The latter was independently corroborated, besides its longer fluorescence lifetime in aqueous solution, through the trends in quenching rate constants of the azoalkanes 1-3 towards electron-deficient versus electron-rich lactone antioxidants of the benzofuranone type. While common inorganic buffer constituents caused no fluorescence quenching, significant quenching was observed, as a curiosity, for hydrogencarbonate (k(q)/10(6) M(-1) s(-1) = 1.7 for 3 versus 2.4 for 1 and 0.45 for 2), with a fully manifested kinetic deuterium isotope effect (k(q)(H(2)O)/k(q)(D(2)O) = 12) for 3. PMID- 20024165 TI - Fluence rate-dependent intratumor heterogeneity in physiologic and cytotoxic responses to Photofrin photodynamic therapy. AB - Photodynamic therapy (PDT) can lead to the creation of heterogeneous, response limiting hypoxia during illumination, which may be controlled in part through illumination fluence rate. In the present report we consider (1) regional differences in hypoxia, vascular response, and cell kill as a function of tumor depth and (2) the role of fluence rate as a mediator of depth-dependent regional intratumor heterogeneity. Intradermal RIF murine tumors were treated with Photofrin PDT using surface illumination at an irradiance of 75 or 38 mW cm(-2). Regional heterogeneity in tumor response was examined through comparison of effects in the surface vs. base of tumors, i.e. along a plane parallel to the skin surface and perpendicular to the incident illumination. 75 mW cm(-2) PDT created significantly greater hypoxia in tumor bases relative to their surfaces. Increased hypoxia in the tumor base could not be attributed to regional differences in Photofrin concentration nor effects of fluence rate distribution on photochemical oxygen consumption, but significant depth-dependent heterogeneity in vascular responses and cytotoxic response were detected. At a lower fluence rate of 38 mW cm(-2), no detectable regional differences in hypoxia or cytotoxic responses were apparent, and heterogeneity in vascular response was significantly less than that during 75 mW cm(-2) PDT. This research suggests that the benefits of low-fluence-rate PDT are mediated in part by a reduction in intratumor heterogeneity in hypoxic, vascular and cytotoxic responses. PMID- 20024167 TI - Demetalation kinetics of natural chlorophylls purified from oxygenic photosynthetic organisms: effect of the formyl groups conjugated directly to the chlorin pi-macrocycle. AB - Demetalation kinetics of natural chlorophyll (Chl) d purified from Acaryochloris marina was first studied and compared with those of Chls a and b. The demetalation rate constant of Chl d, which possessed a formyl group at the 3 position, was five-fold smaller than that of Chl a possessing a vinyl group at the same position in aqueous acetone at the proton concentration of 1.2 x 10(-3) M at 25 degrees C. In contrast, the demetalation rate constant of Chl b possessing a formyl group at the 7-position was 26 times smaller than that of Chl a. The activation energy of demetalation reaction of Chl d was larger than that of Chl a, but smaller than that of Chl b. These indicate that the substitution effect of 3-formyl group on the acidic removal of central magnesium in Chls was smaller than that of 7-formyl group. PMID- 20024168 TI - Encapsulation of 2-(4'-N,N-dimethylamino)phenylimidazo[4,5-b]pyridine in beta cyclodextrin: effect on H-bond-induced intramolecular charge transfer emission. AB - The effect of beta-cyclodextrin (beta-CD) inclusion complex formation on the hydrogen bond-induced intramolecular charge transfer (ICT) of 2-(4'-N,N dimethylamino)phenylimidazo[4,5-b]pyridine (DMAPIP-b) has been examined by fluorescence excitation, emission and time-resolved fluorescence techniques. The study reveals that DMAPIP-b forms 1 : 1 inclusion complex with beta-CD. The host guest complex is formed by partial inclusion of DMAPIP-b, i.e. only the dimethylaminophenyl ring is encapsulated inside the core of the beta-CD nanocavity. The imidazopyridine ring of the guest molecule resides outside CD cavity and forms H-bonds with the water molecules that are present near the rim and in the bulk phase. (1)H NMR studies are used to confirm the inclusion complex. The H-bond of water with the pyridine nitrogen ensures the formation of the ICT state and both normal and ICT emissions are enhanced inside the beta-CD cavity. Fluorescence lifetime measurements suggest that the formation of the ICT state from the locally excited state is irreversible. Dual emission is observed in the presence of beta-CD at pH approximately 3.5, due to emission from monocations formed by the protonation of pyridine nitrogen (MC1) and imidazole nitrogen (MC2). PMID- 20024169 TI - Drug efflux transporters, MRP1 and BCRP, affect the outcome of hypericin-mediated photodynamic therapy in HT-29 adenocarcinoma cells. AB - Photodynamic therapy (PDT) is a flexible multi-target therapeutic approach. One of the main requirements of successful PDT is sufficient intracellular concentration of an applicable photosensitizer. Mechanisms of anticancer drug elimination by tumour cells are mostly linked to the elevated expression and activity of P-glycoprotein (P-gp), multidrug resistance-associated protein 1 (MRP1), breast cancer resistance protein (BCRP) and P450 monooxygenases. The interaction of hypericin with this cell drug-defence system is still unclear. We report here for the first time increased activity of MRP1 and BCRP in HT-29 colon cancer cells treated with hypericin per se. On the contrary, pre-treatment with proadifen (SKF525A) affected the function of MRP1 and BCRP leading to increased hypericin content, which might indicate a possible link between proadifen and these ABC transporter proteins. Subsequent enhanced intracellular oxidative stress was accompanied by loss of mitochondrial membrane potential, activation of caspase-9 and -3, PARP cleavage and onset of apoptosis. In conclusion, our study suggests that drug efflux transporters MRP1 and BCRP affect the pharmacokinetics of hypericin in HT-29 colon adenocarcinoma cells, and the action of hypericin mediated PDT (HY-PDT) should be modulated by pre-treatment with their specific inhibitors. PMID- 20024170 TI - Photophysical properties and photodynamic activity of octacationic oxotitanium(IV) phthalocyanines. AB - The photophysical and photosensitizing properties of two octacationic oxotitanium phthalocyanines (TiOPcs), bearing pyridiniomethyl or cholinyl substituents, have been studied in aqueous and alcohol solutions. In water, both compounds were monomeric with the high quantum yields of fluorescence (Phi(F) = 0.17-0.19) and singlet oxygen formation (Phi(Delta) = 0.4-0.5). The Phi(F) and Phi(Delta) of both phthalocyanines decreased with the increase of solvent hydrophobicity from water to ethanol. This effect was much stronger in alcohol solutions of the pyridiniomethyl-substituted phthalocyanine and probably results from aggregation of TiOPc molecules caused by association of chloride anions with phthalocyanine cationic groups. Evidence is presented that under illumination aqueous TiOPc solutions also produce hydroxyl radicals, which probably appear owing to photocleavage of water molecules. The quantum yield of OH formation was (3-5) x 10(-5) after argon purging and twice as much in the presence of air. It is shown that irradiation of TiOPc solutions causes photobleaching of TiOPcs. The photobleaching quantum yield in water was found to be about 1 x 10(-4). The data suggest that photobleaching occurs owing to the reactivity of hydroxyl radicals, though singlet oxygen is generated by TiOPcs much more efficiently. The phototoxicity of the tested TiOPcs toward bacteria has been revealed. It is proposed that both OH and (1)O(2) might be responsible for the observed bactericidal effects. PMID- 20024171 TI - Photochemical investigation of a photochromic diarylethene compound that can be used as a wide range actinometer. AB - The photochromic diarylethene derivative 1,2-bis(5-(4-ethynylphenyl)-2 methylthiophen-3-yl)perfluorocyclopentene (1) was submitted to photochemical, thermal stability and fatigue resistance studies in acetonitrile, also to evaluate its possible application as a new actinometer. This photochromic system covers a wide spectral absorption range, with intense bands in the UV and visible regions for the open-ring and closed-ring isomers, respectively. Very high ring closure quantum yield values were obtained, in contrast with the low ring-opening quantum yields, which are nevertheless high enough to exploit 1 as an actinometer. The procedure required to determine the photon flux of an irradiation source with this fatigue resistant compound is indeed very simple. PMID- 20024172 TI - Cucurbituril complexes cross the cell membrane. AB - Cucurbiturils (CBs) of the appropriate size (CB[7] and CB[8]) form strong guest host complexes in phosphate buffer solution (PBS) with acridine orange (AO) and pyronine Y (PYY) with 1 : 1 and 2 : 1 stoichiometries for CB[7] and CB[8] complexes, respectively. Binding constants in the range 0.87-1.60 x 10(6) M(-1) and 5.2-6.3 x 10(13) M(-2) were determined by titration with fluorescence spectroscopy for 1 : 1 and 2 : 1 complexes, respectively. These binding constants in PBS and the eight-fold excess of CBs minimize the presence of free dye in solution and also stabilize the host-guest complex in the culture medium. Images showing that the CB complexes can cross the cell membrane of 3T3 cells have been acquired using fluorescence microscopy. Given the current importance of supramolecular CB complexes and the search for new drug delivery systems, the present findings open avenues for the use of CBs as nanocapsules to transport drugs into the cells. PMID- 20024173 TI - Pyrearinus termitilluminans larval click beetle luciferase: active site properties, structure and function relationships and comparison with other beetle luciferases. AB - Several beetle luciferases have been cloned and sequenced. However, most studies on structure and function relationships and bioanalytical applications were done with firefly luciferases, which are pH sensitive. Several years ago we cloned Pyrearinus termitilluminans larval click beetle luciferase, which displays the most blue-shifted bioluminescence among beetle luciferases and is pH insensitive. This enzyme was expressed in E. coli, purified, and its properties investigated. This luciferase shows slower luminescence kinetics, K(M) values comparable to other beetle luciferases and high catalytic constant. Fluorescence studies with 8 anilino-1-naphtalene-sulfonic acid (1,8-ANS) and modeling studies suggest that the luciferin binding site of this luciferase is very hydrophobic, supporting the solvent and orientation polarizability effects as determining mechanisms for bioluminescence colors. Although pH insensitive in the range between pH 6-8, at pH 10 this luciferase displays a remarkable red-shift and broadening of the bioluminescence spectrum. Modeling studies suggest that the residue C312 may play an important role in bioluminescence color modulation. Compared to other beetle luciferases, Pyrearinus termitilluminans luciferase also displays higher thermostability and sustained luminescence in a bacterial cell environment, which makes this luciferase particularly suitable for in vivo cell analysis and bioimaging. PMID- 20024176 TI - Viruses and their potential in bioimaging and biosensing applications. AB - Successful development of ultrasensitive constructs for bioimaging and biosensing is a challenging task. Recently, viruses have drawn increasing attention due to their exquisite three-dimensional structures and unique properties, including multivalency, orthogonal reactivities, and responsiveness to genetic modifications. With such well-characterized structures, functional units, such as imaging and binding motifs, can be engineered on the surface of viruses in a programmable, polyvalent manner, which leads to novel nanosized sensing/imaging systems with enhanced signaling and targeting performance. This review highlights some recent progress in the applications of viruses in bioimaging and biosensing. PMID- 20024177 TI - Molecular imaging of adrenal gland by desorption electrospray ionization mass spectrometry. AB - Subtle differences in the spatial distributions of closely related compounds including norepinephrine and epinephrine as well multiple lipids are easily distinguished in adult porcine (17 x 8 mm) and rabbit (7 x 4 mm) adrenal glands in a DESI-MS imaging experiment at atmospheric pressure with a spatial resolution of approximately 200 microm. Sensitive and specific detection in the course of DESI imaging discloses details of catecholamine distribution in porcine adrenal medulla and cortex; the average mass of epinephrine interrogated in each pixel is estimated to be about 150 pg. The distribution of ascorbic acid was revealed in the negative ion mode. In addition, the distribution of cholesterol, which cannot be observed using conventional DESI, was obtained using in situ reaction with betaine aldehyde added to the DESI spray while imaging the porcine adrenal gland tissue. Four characteristic types of distributions were observed, with major amounts of the components in the medulla, the cortex, the reticular zone or in the fourth case, being homogeneously distributed. The results agree with and extend information available from histological studies. PMID- 20024178 TI - A novel reversible relative-humidity indicator ink based on methylene blue and urea. AB - A new relative-humidity sensitive ink based on methylene blue and urea is described which can utilise the deliquescent nature of urea. PMID- 20024179 TI - Improving limits of detection for B-type natriuretic peptide using PC-IDMS: an application of the ALiPHAT strategy. AB - Hydrophobic tagging of biomolecules has been reported by our group and others to increase their ionization efficiency during electrospray ionization and facilitate their detection by mass spectrometry. As such, hydrophobic tagging should provide a viable method for augmenting MS-based quantification of low abundance proteins by decreasing their detection limits. Herein we have evaluated two commercial alkylation reagents and several newly synthesized hydrophobic alkylation reagents for their utility in quantifying B-type Natriuretic Peptide, a low abundance cardiac biomarker, by protein cleavage isotope dilution mass spectrometry. For the cysteine containing tryptic peptide evaluated, a approximately 3.5-fold decrease in the detection limit was observed for the best performing hydrophobic reagent, 2-iodo-N-octylacetamide, relative to the commonly used alkylation reagent, iodoacetamide. Additionally, we have evaluated the use of nonpolar surface areas as a metric for assessing the effectiveness of the alkylation reagents in improving ESI response. PMID- 20024180 TI - Bioconjugated lanthanide luminescent helicates as multilabels for lab-on-a-chip detection of cancer biomarkers. AB - The lanthanide binuclear helicate [Eu(2)(L(C2(CO(2)H)))(3)] is coupled to avidin to yield a luminescent bioconjugate EuB1 (Q = 9.3%, tau((5)D(0)) = 2.17 ms). MALDI/TOF mass spectrometry confirms the covalent binding of the Eu chelate and UV-visible spectroscopy allows one to determine a luminophore/protein ratio equal to 3.2. Bio-affinity assays involving the recognition of a mucin-like protein expressed on human breast cancer MCF-7 cells by a biotinylated monoclonal antibody 5D10 to which EuB1 is attached via avidin-biotin coupling demonstrate that (i) avidin activity is little affected by the coupling reaction and (ii) detection limits obtained by time-resolved (TR) luminescence with EuB1 and a commercial Eu-avidin conjugate are one order of magnitude lower than those of an organic conjugate (FITC-streptavidin). In the second part of the paper, conditions for growing MCF-7 cells in 100-200 microm wide microchannels engraved in PDMS are established; we demonstrate that EuB1 can be applied as effectively on this lab-on-a-chip device for the detection of tumour-associated antigens as on MCF-7 cells grown in normal culture vials. In order to exploit the versatility of the ligand used for self-assembling [Ln(2)(L(C2(CO(2)H)))(3)] helicates, which sensitizes the luminescence of both Eu(III) and Tb(III) ions, a dual on-chip assay is proposed in which estrogen receptors (ERs) and human epidermal growth factor receptors (Her2/neu) can be simultaneously detected on human breast cancer tissue sections. The Ln helicates are coupled to two secondary antibodies: ERs are visualized by red-emitting EuB4 using goat anti-mouse IgG and Her2/neu receptors by green-emitting TbB5 using goat anti-rabbit IgG. The fact that the assay is more than 6 times faster and requires 5 times less reactants than conventional immunohistochemical assays provides essential advantages over conventional immunohistochemistry for future clinical biomarker detection. PMID- 20024181 TI - Fourier transform infrared (FTIR) spectromicroscopic characterization of stem like cell populations in human esophageal normal and adenocarcinoma cell lines. AB - We have tested an approach to identify putative cancer stem cells that involves measurement of the infrared absorption spectrum of individual cells in an aqueous environment, and their subsequent classification using multivariate data analysis techniques. Two primary esophageal cell lines were characterized: the immortalized normal esophageal epithelial cell line, Het-1A, and the esophageal adenocarcinoma cell line, OE33. In addition, we also evaluated spheroids, reflecting stem-like cell populations, which were derived from each parent cell line when grown in serum-free media. As differences in cell size appeared to be a strong discriminating factor, a correction needs to be performed to allow a reliable classification based on infrared absorption spectra. We demonstrated that stem-like cells derived from Het-1A could easily be discriminated on the basis of absorbance differences in the 1000-1200 cm(-1) spectral interval, whereas this was not possible for OE33. Furthermore, we found that changes due to aging of OE33 cells in culture dominated the infrared absorption spectra and somewhat limited the potential of this approach to identify stem-like cell populations using this in vitro model system. PMID- 20024183 TI - Potassium-sensitive G-quadruplex DNA for sensitive visible potassium detection. AB - A novel visual method for K(+) detection is developed utilizing a K(+)-sensitive G-quadruplex DNA named PS5.M as the sensing element. In the absence of K(+), PS5.M is in a random coil state and does not bind hemin. Upon addition of K(+), PS5.M folds into the quadruplex structure stabilized by K(+). Such quadruplex structure is able to bind hemin to form the hemin-G-quadruplex DNAzyme that catalyzes the H(2)O(2)-mediated oxidation of colorless 3,3',5,5' tetramethylbenzidine to the blue product. Under the optimal conditions, the color change can be clearly observed with the naked eye in the concentration range from 2 to 1000 microM. Through this facile approach, K(+) can be detected down to 2 microM, with high selectivity against other cations. In view of its simplicity, sensitivity and specificity, our developed visual method for K(+) detection would potentially be utilized for bioassays and K(+)-sensitive test paper. PMID- 20024182 TI - Sequential CD34 cell fractionation by magnetophoresis in a magnetic dipole flow sorter. AB - Cell separation and fractionation based on fluorescent and magnetic labeling procedures are common tools in contemporary research. These techniques rely on binding of fluorophores or magnetic particles conjugated to antibodies to target cells. Cell surface marker expression levels within cell populations vary with progression through the cell cycle. In an earlier work we showed the reproducible magnetic fractionation (single pass) of the Jurkat cell line based on the population distribution of CD45 surface marker expression. Here we present a study on magnetic fractionation of a stem and progenitor cell (SPC) population using the established acute myelogenous leukemia cell line KG-1a as a cell model. The cells express a CD34 cell surface marker associated with the hematopoietic progenitor cell activity and the progenitor cell lineage commitment. The CD34 expression level is approximately an order of magnitude lower than that of the CD45 marker, which required further improvements of the magnetic fractionation apparatus. The cells were immunomagnetically labeled using a sandwich of anti CD34 antibody-phycoerythrin (PE) conjugate and anti-PE magnetic nanobead and fractionated into eight components using a continuous flow dipole magnetophoresis apparatus. The CD34 marker expression distribution between sorted fractions was measured by quantitative PE flow cytometry (using QuantiBRITE PE calibration beads), and it was shown to be correlated with the cell magnetophoretic mobility distribution. A flow outlet addressing scheme based on the concept of the transport lamina thickness was used to control cell distribution between the eight outlet ports. The fractional cell distributions showed good agreement with numerical simulations of the fractionation based on the cell magnetophoretic mobility distribution in the unsorted sample. PMID- 20024184 TI - High throughput screening of lead utilising disposable screen printed shallow recessed microelectrode arrays. AB - The cathodic stripping voltammetry of lead at disposable screen printed shallow recessed microelectrode arrays has been developed for the first time. The array comprises 6 microdiscs which have radii of 116 (+/-6) microns which are recessed by 4 microns and are separated by 2500 microns from their nearest neighbour in a hexagonal arrangement. The electroanalytical determination of lead was explored in 0.1 M nitric acid and found that using a 120 s deposition time, a detection limit of 3 microM is feasible which is not possible utilising a screen printed graphite macro-electrode. The sensitivity of this analytical protocol can be tailored by varying the deposition time and it is found that increasing this to 320 s facilitates a limit of detection of 39 nM. This methodology is shown to be feasible for the portable and economical screening of lead in river water samples at the levels indicated by the EC Dangerous Substances Directive (76/464/EEC). PMID- 20024185 TI - Rapid identification of stabilisers in polypropylene using time-of-flight mass spectrometry and DART as ion source. AB - A method for analysing plastic samples without any sample pretreatment using direct analysis in real time mass spectrometry (DART-MS) was developed. DART-MS allows the direct, simple and rapid identification of polymer additives in plastic products. To demonstrate the suitability of DART-MS for the detection of a wide range of commonly employed stabilising agents, a test set of 21 stabilisers was selected. In a first step standard solutions of these stabilisers in toluene as well as toluene-extracts from polymer samples were analysed. Subsequently, to prove the applicability of the developed DART-MS method also for the direct analysis of plastic products, samples of polypropylene containing a range of stabilisers were prepared using a lab-scale compounder. Polymer samples were cut into 0.5 cm wide pieces and directly placed into the DART ion source. Focusing on the DART ionisation, several parameters like discharge needle potential, potential of the grid electrode and the discharge electrode, the heater temperature and the gas flow had to be varied to guarantee optimum results. Both positive and negative ionisation was tested, whereby the positive ion mode led to higher signal intensities for all analytes. Determination of accurate masses to improve the certainty in signal assignment could be achieved by using PEG 600 as an internal standard for mass calibration. The developed method allowed the detection of all selected additives (including some of their degradation products) in real polymer samples. PMID- 20024186 TI - Cadmium binding in mixtures of phytochelatins and their fragments: a voltammetric study assisted by multivariate curve resolution and mass spectrometry. AB - Phytochelatins (PC(n), (gamma-Glu-Cys)(n)Gly) are cysteine-rich peptides synthesized by plants which are involved in metal bioregulation and phytoremediation. Multivariate Curve Resolution by Alternating Least Squares (MCR ALS) is applied to voltammetric data obtained from the analysis of the competitive binding of Cys or Cys-Gly with PC(2) or PC(3) by Cd(2+). The displacements between ligands, the chain length dependence on the competitive binding to PC(n) and the possible existence of mixed ligand metal-complexes are investigated. The shape analysis of the resulting pure voltammograms and concentration profiles of the components resolved by MCR-ALS suggests that ligands containing more thiol groups are able to displace the shorter chain ligands from their metal complexes, whereas the opposite does not happen. Electrochemical results are compared with ESI-MS measurements. PMID- 20024188 TI - Fluorescent sensing of homocysteine in urine: using fluorosurfactant-capped gold nanoparticles and o-Phthaldialdehyde. AB - This study reports the development of a simple, sensitive, and selective detection system for homocysteine (HCys) based on the combination of fluorosurfactant-capped gold nanoparticles (FSN-AuNPs) and o-Phthaldialdehyde (OPA). The proposed assay utilizes FSN-AuNPs as extractors for HCys and cysteine (Cys), which can then be collected by centrifugation. As long as the HCys and Cys are isolated from the initial sample, they can be liberated from the NP surface by 2-mercaptoethanol (2-ME). The derivatization of released HCys with OPA/2-ME has a strong fluorescence maximum at 485 nm, whereas the derivatization of released Cys with the same reagent shows an extremely weak fluorescence maximum at 457 nm. As a result, the selectivity of this system is more than 100-fold for HCys over any aminothiols when excited at 370 nm. The extraction and derivation efficiencies are monitored as functions of the concentration of FSN-AuNPs and OPA, respectively. The proposed system has a detection limit of 180 nM at a signal-to-noise ratio of 3 for HCys. This study validates the applicability of this system by analyzing the amount of HCys in urine samples. PMID- 20024187 TI - Dual contactless conductivity and amperometric detection on hybrid PDMS/glass electrophoresis microchips. AB - A new approach for the integration of dual contactless conductivity and amperometric detection with an electrophoresis microchip system is presented. The PDMS layer with the embedded channels was reversibly sealed to a thin glass substrate (400 microm), on top of which a palladium electrode had been previously fabricated enabling end-channel amperometric detection. The thin glass substrate served also as a physical wall between the separation channel and the sensing copper electrodes for contactless conductivity detection. The latter were not integrated in the microfluidic device, but fabricated on an independent plastic substrate allowing a simpler and more cost-effective fabrication of the chip. PDMS/glass chips with merely contactless conductivity detection were first characterized in terms of sensitivity, efficiency and reproducibility. The separation efficiency of this system was found to be similar or slightly superior to other systems reported in the literature. The simultaneous determination of ionic and electroactive species was illustrated by the separation of peroxynitrite degradation products, i.e. NO(3)(-) (non-electroactive) and NO(2)( ) (electroactive), using hybrid PDMS/glass chips with dual contactless conductivity and amperometric detection. While both ions were detected by contactless conductivity detection with good efficiency, NO(2)(-) was also simultaneously detected amperometrically with a significant enhancement in sensitivity compared to contactless conductivity detection. PMID- 20024189 TI - BSA activated CdTe quantum dot nanosensor for antimony ion detection. AB - A novel fluorescent nanosensor for Sb(3+) determination was reported based on thioglycolic acid (TGA)-capped CdTe quantum dot (QD) nanoparticles. It was the first antimony ion sensor using QD nanoparticles in a receptor-fluorophore system. The water-soluable TGA-capped CdTe QDs were prepared through a hydrothermal route, NaHTe was used as the Te precursor for CdTe QDs synthesis. Bovine serum albumin (BSA) conjugated to TGA-capped CdTe via an amide link interacting with carboxyl of the TGA-capped CdTe. When antimony ion enters the BSA, the lone pair electrons of the nitrogen and oxygen atom become involved in the coordination, switching off the QD emission and a dramatic quenching of the fluorescence intensity results, allowing the detection of low concentrations of antimony ions. Using the operating principle, the antimony ion sensor based on QD nanoparticles showed a very good linearity in the range 0.10-22.0 microg L(-1), with the detection limit lower than 2.94 x 10(-8) g L(-1) and the relative standard deviation (RSD) 2.54% (n = 6). In a study of interferences, the antimony sensitive TGA-QD-BSA sensor showed good selectivity. Therefore, a simple, fast, sensitive, and highly selective assay for antimony has been built. The presented method has been applied successfully to the determination of antimony in real water samples (n = 6) with satisfactory results. PMID- 20024190 TI - A sensitive electrochemical approach for monitoring the effects of nano-Al2O3 on LDH activity by differential pulse voltammetry. AB - In this paper, a sensitive electrochemical approach for monitoring the effect of nano-Al(2)O(3) on lactate dehydrogenase (LDH) activity is established. It is based on the determination of reduction current of NAD(+) involved in enzyme promoting catalytic reaction of "pyruvate + NADH + H+ [see text]L D H lactate + NAD+" by differential pulse voltammetry (DPV). Various influencing factors including nano-type, nano-size, and adsorbed pollutant organics have been investigated. The experimental results show that the proposed electrochemical method is useful in monitoring and evaluating the toxic effects of nanoparticles, which might be suitable to the environmental pollutant's toxicity analysis. PMID- 20024191 TI - An electrochemical molecular switch for one-step, reusable detection of a single base mutation of DNA. AB - A new strategy for one-step, reusable and sensitive detection of a single-base mutation based on an electrochemical molecular switch is developed in the present work. When the hybridization reaction takes place in the presence of target DNA, the Fc-labeled terminal of the open switch molecule can be captured by the probe through the predesigned complementary bases of both sequences. By this method, a signal-on sensor featuring both generalizability and simplicity towards reagentless detection of DNA with sensitivity and selectivity electrochemical system is built on. The approach had been demonstrated with the identification of a single-base mutation of alpha-thalassemia point mutation in Hb Constant Spring codon 142 (TAA --> CAA). The wild-type and mutant-type of the synthetic 16 mer DNA sequences as the model targets were successfully discriminated. The results showed that the response signal was linear to the logarithm of the target concentration in the range from 0.01 to 100 pM with a detection limit of 0.01 pM. The regeneration experiment demonstrated that the sensor interface can be easily and successfully regenerated. All these revealed that the present system is a promising candidate for single-base mutation discrimination. PMID- 20024192 TI - Analytical protocol for identification of BMAA and DAB in biological samples. AB - Beta-N-methylamino-L-alanine (BMAA) is a non-protein amino acid, thought to be inflicting neurodegenerative diseases related to ALS/PDC in human beings. Due to conflicting data concerning the presence of BMAA in various biological matrixes, we present a robust and sensitive method for high confidence identification of BMAA after derivatization by 6-aminoquinolyl-N-hydroxysuccinimidyl carbamate (AQC). The efficient sample pretreatment in combination with LC-MS/MS SRM enables chromatographic separation of BMAA from the isomer 2,3-diaminobutyric acid (DAB). The method is applicable for selective BMAA/DAB detection in various biological samples ranging from a prokaryotic cyanobacterium to eukaryotic fish. PMID- 20024193 TI - Following interfacial kinetics in real time using broadband evanescent wave cavity-enhanced absorption spectroscopy: a comparison of light-emitting diodes and supercontinuum sources. AB - A white light-emitting diode (LED) with emission between 420 and 700 nm and a supercontinuum (SC) source with emission between 450 and 2500 nm have been compared for use in evanescent wave broadband cavity-enhanced absorption spectroscopy (EW-BB-CEAS). The method is calibrated using a dye with known absorbance. While the LED is more economic as an excitation source, the SC source is superior both in terms of baseline noise (noise equivalent absorbances lower than 10(-5) compared to 10(-4) absorbance units (a.u.)) and accuracy of the measurement; these baseline noise levels are comparable to evanescent wave cavity ringdown spectroscopy (EW-CRDS) studies while the accessible spectral region of EW-BB-CEAS is much larger (420-750 nm in this study, compared to several tens of nanometres for EW-CRDS). The improvements afforded by the use of an SC source in combination with a high sensitivity detector are demonstrated in the broadband detection of electrogenerated Ir(IV) complexes in a thin-layer electrochemical cell arrangement. Excellent signal to noise is achieved with 10 micros signal accumulation times at a repetition rate of 600 Hz, easily fast enough to follow, in real time, solution kinetics and interfacial processes. PMID- 20024194 TI - A novel isotachophoresis of cobalt and copper complexes by metal ion substitution reaction in a continuous moving chelation boundary. AB - A novel separation mode of isotachophoresis (ITP) was advanced for the study on the continuous moving chelation boundary (MCB) formed with EDTA and two metal ions of Co(II) and Cu(II). The experiments were performed systemically. The relevant results indicated that: (1) there were three boundaries in the whole system, viz., a sharp MCB, a wide moving substitution boundary (MSB) and a sharp complex boundary (CB); (2) within the MSB, an ion substitution reaction occurred between [Co-EDTA](2-) and Cu(II), and the reaction resulted in the release of Co(II) and EDTA from [Co-EDTA](2-) and the binding of Cu(II) with the released EDTA due to log K(Cu(II)) (= 18.80) > log K(Co(II)) (= 16.31); (3) because of the novel ITP mode induced by the MSB as well as the merging of the MCB and CB, the original low concentration Co(II) and Cu(II) were chemically separated as two characteristic coloured zones of pink [Co-EDTA](2-) and blue [Cu-EDTA](2-), and the sensitivities for detection of the two metal ions were greatly enhanced. The quantitative analyses of the zone composition by ICP-AES and UV-vis spectrophotometry supported the mechanism of the novel separation mode induced by the MSB. The further theoretical and experimental results indicated that the separation mode was a novel ITP relied on moving reaction boundary (MRB), rather than a classic ITP based on the moving boundary system developed about 60 years ago. These findings provide guidance for the development of the MRB and the MCB based ITP separation of metal ions in environmental and biological matrices. PMID- 20024195 TI - Functionalized monolayers on mesoporous silica and on titania nanoparticles for mercuric sensing. AB - Heterogeneous "naked-eye" colorimetric and spectrophotometric cation sensors were prepared by immobilization of an azobenzene-coupled receptor onto mesoporous silica (AR-SiO(2)) or titania nanoparticles (AR-TiO(2)) via sol-gel or hydrolysis reactions. The optical sensing ability of AR-SiO(2) was studied by addition of metal ions such as K(+), Ca(2+), Sr(2+), Co(2+), Cd(2+), Pb(2+), Zn(2+), Fe(3+), Cu(2+) and Hg(2+) ions (all as chlorides) in aqueous solution. Upon the addition of Hg(2+) ion in suspension, the AR-SiO(2) resulted in a color change from yellow to deep red. No significant color changes were observed in the parallel experiments with K(+), Ca(2+), Sr(2+), Co(2+), Cd(2+), Pb(2+), Zn(2+), Fe(3+) or Cu(2+) ion. These findings confirm that the AR-SiO(2) can be useful as chemosensors for selective detection of Hg(2+) ion over a range of metal ions in aqueous solution. Also, the color change of AR-SiO(2) was independent of the presence of anions NO(3)(-), ClO(4)(-), Br(-) and I(-). We also prepared a portable chemosensor kit by coating a 4 microm thick film of AR-TiO(2) onto a glass substrate. We found that this AR-TiO(2) film detects Hg(2+) ion at pH 7.4 with a sensitivity of 28 nM. Finally, we tested the effect of pH on AR-TiO(2) with Hg(2+) ion between pH 1.0 to 11.0. The absorbance and color changes of AR TiO(2) were almost constant between pH 4 and 11. The results imply that the AR TiO(2) film is applicable as a portable colorimetric sensor for the detection of Hg(2+) ion in the environmental field. PMID- 20024196 TI - The differentiation and engraftment potential of mouse hematopoietic stem cells is maintained after bio-electrospray. AB - The bio-electrospray technique has been recently pioneered to manipulate living, immortalised and primary cells, including a wide range of stem cells. Studies have demonstrated that the creation of viable, fully functional in vitro microenvironments is possible using this technique. By modifying the bio electrospray procedure (referred to as cell electrospinning), a variety of microenvironment morphologies have been fabricated. Because bio-electrospraying of biological material is a relatively new technique, it is important to determine if there are any unwanted consequences to the manipulated cells as a result of the procedure. Here, we establish the validity of the process using a heterogeneous, living population of hematopoietic stem/progenitor cells, using a functional in vitro assay and in vivo mouse model to investigate for side-effects that previous in vitro assays may not have detected. Our studies demonstrate that these bio-protocols have no obvious negative effects, thus indicating significant promise for utility in biological sciences and for a plethora of healthcare applications. PMID- 20024198 TI - Cyclic voltammetry on recessed nanodisk-array electrodes prepared from track etched polycarbonate membranes with 10-nm diameter pores. AB - This paper described cyclic voltammograms (CVs) on a recessed nanodisk-array electrode (RNE) prepared from a track-etched polycarbonate membrane (TEPCM) with 10-nm diameter pores. A RNE was prepared via sputtering of a gold thin film onto a TEPCM whose pores were coated with polyvinylpyrrolidone. CVs of such a RNE changed from peak-shaped to sigmoidal with decreasing scan rate due to the transition of diffusion modes of redox-active molecules, as with previous reports. However, faradic currents in these CVs were larger than theoretical currents calculated from the membrane thickness, pore density, and pore diameter. The larger faradic currents may reflect the presence of a surface layer on TEPCM nanopores to which the redox species preferentially distribute as compared with the nanopore cavity. The limiting current of uncharged 1,1'-ferrocenedimethanol was not affected by solution pH or supporting electrolyte concentration. In contrast, those of anionic ferricyanide and cationic ferrocenylmethyltrimethylammonium decreased and increased, respectively, with increasing pH from 4 to 9. These pH-dependent changes in limiting current were larger at lower supporting electrolyte concentration. The dependence of the CVs on solution pH and supporting electrolyte concentrations was attributed to electrostatic interaction between the redox species and the charged TEPCM nanopores. PMID- 20024197 TI - Rapid tandem-column micro-gas chromatography based on optofluidic ring resonators with multi-point on-column detection. AB - We demonstrated a novel tandem-column micro-gas chromatography (microGC) based on optofluidic ring resonator (OFRR). The OFRR is a thin-walled fused silica capillary whose interior surface is coated with a polymeric stationary phase. The circular cross section of the OFRR forms the micro-ring resonator and supports whispering gallery modes (WGMs). Via tapered optical fibers in contact with the OFRR, the WGM can be excited externally at any positions along the OFRR capillary, thus enabling multi-point, on-column, real-time detection of vapor molecules flowing through the OFRR. In the present OFRR-based tandem-column-based microGC implementation, a 180 cm long conventional GC column coated with a nonpolar stationary phase was followed by a relatively short OFRR column coated with a polar phase. Two detection positions, one at the inlet of the OFRR and the other a few centimeters downstream, were used to monitor the separation achieved by the first and the second column, respectively. Owing to the multi-point on column detection that provides complementary retention time information on each chemical compound, co-eluted analytes can be well separated and identified on at least one detection channel and no modulation is needed at the interface of tandem columns. Separation and detection of twelve analytes with various volatilities and polarities within four minutes were demonstrated. In addition, the chromatograms obtained from three different locations along the OFRR column demonstrated the system's capability of on-column monitoring of the separation process for the target analyte in a vapor mixture. Our results will lead to the development of a rapid, simple, and portable microGC system with significantly improved selectivity and chemical identification capabilities. PMID- 20024199 TI - Multiplex tumor marker detection with new chemiluminescent immunoassay based on silica colloidal crystal beads. AB - A new multiplex chemiluminescent immunoassay (CLIA) based on silica colloidal crystal beads (SCCBs) was developed for tumor marker detection. As the code is the characteristic reflection peak originating from the stop-band of colloid crystal, they avoid photobleaching, the potential interference of encoding fluorescence with analyte-detection fluorescence and chemical instability. Meanwhile our SCCBs suspension array improved the luminescence analysis efficiency by using chemiluminescent detection of enzyme labels. By forming a sandwich immunocomplex on SCCBs, the proposed suspension array was used for simultaneous multiplex detection of tumor markers in one test tube. The results showed that the linear range was 0.5-100ng ml(-1) and 1.0-120ng ml(-1) for carcinoembryonic antigen (CEA) and alpha-fetoprotein (AFP) with a detection limit of 0.12ng ml(-1) and 0.16ng ml(-1) at 3sigma. The proposed array showed the storage stability and the accuracy for sample detection were acceptable, and the results were in acceptable agreement with the reference electrochemiluminescence method. This technique provided an automated, simple, sensitive and low-cost approach for multianalyte immunoassay. PMID- 20024200 TI - Development and validation of a multiplex bead assay for measuring growth mediators in wound fluid. AB - Large amounts of biological samples are usually required to measure multiple components by the enzyme-linked immunosorbant assay. However, the amounts of many tissue extracts and fluids, including gingival crevicular fluid (GCF), are generally extremely small. The aim of this study was, therefore, to develop and validate a novel multiplex bead assay (MBA) to simultaneously measure a profile of healing-related mediators in the GCF of treated periodontal wounds. An MBA was developed and validated by assessment of assay selectivity, recovery, precision and sensitivity, using eight recombinant human growth mediators as assay standards. GCF samples were collected on paper strips from healing wound (test) and healthy unaffected (control) sites of 15 patients with periodontitis, seven days post-periodontal surgery. Each GCF sample was eluted and the levels of the mediators measured using the MBA and antibody pairs specific for angiopoietin-1, vascular endothelial growth-factor, bone morphogenetic protein-2, osteoprotegerin, tissue inhibitor of metalloprotease-1 (TIMP-1), basic fibroblast growth-factor, keratinocyte growth-factor, and platelet derived growth-factor. Less than 1.8% of cross-reactivity was observed between antibodies and the eight different analytes, for which the recovery was more than 85%. Mean intra- and inter-assay precision were within the acceptance criteria of 20% and 25%, respectively. Detection of all mediators was highly sensitive (DMT and c,a-DMT<>DMT leads to the formation of dimer radical anions with the lifetime of at least 80 micros, thus showing that the latter are much more stable than previously believed. PMID- 20024206 TI - Highly enantioselective hetero-Diels-Alder reactions between Rawal's diene and aldehydes catalyzed by chiral dirhodium(II) carboxamidates. AB - The first example of a chiral Lewis acid-catalyzed enantioselective hetero-Diels Alder (HDA) reaction between 1-dimethylamino-3-silyloxy-1,3-butadiene (Rawal's diene) and aldehydes is described. The cycloaddition reaction under the influence of 1 mol% of dirhodium(II) tetrakis[N-benzene-fused-phthaloyl-(S) piperidinonate], Rh(2)(S-BPTPI)(4), proceeded cleanly and gave, after treatment with acetyl chloride, the corresponding dihydropyranones in up to 99% ee. PMID- 20024207 TI - Highly enantioselective synthesis of tertiary alcohols: C2-symmetric N,N'-dioxide Sc(III) complex promoted direct aldol reaction of alpha-ketoesters and diazoacetate esters. AB - A C(2)-symmetric N,N'-dioxide-Sc(III) complex has been developed to promote the asymmetric catalytic aldol reaction of alpha-ketoesters and diazoacetate esters to afford tertiary alcohols in good yields with excellent enantioselectivities. PMID- 20024208 TI - CO dissociation induced by adsorbed oxygen and water on Ir(111). AB - Although CO does not dissociate on the clean Ir(111) surface, the addition of atomic oxygen induces CO dissociation at low temperatures (lower than 400 K); similarly, CO dissociation has also been observed on water co-adsorbed Ir(111) or water and oxygen co-adsorbed Ir(111). PMID- 20024209 TI - A supramolecular hydrogel as a reusable heterogeneous catalyst for the direct aldol reaction. AB - An L-proline based supramolecular hydrogel is used as an efficient heterogeneous organocatalyst for the direct aldol reaction with high stereoselectivity (up to 90% ee) and recyclability (up to 3 runs). The reversible nature of this self assembled supramolecular system allows for easy recovery and regeneration of the catalyst. PMID- 20024210 TI - Metamagnetism in a pi-stacked bis-dithiazolyl radical. AB - At temperatures below 5 K the field dependence of the magnetization of the pi stacked bis-dithiazolyl radical 1 (R(1) = Me, R(2) = H) is consistent with metamagnetic behavior. PMID- 20024211 TI - Facile synthesis of a chiral urea bridged bisoxazoline ligand and structural characterization of its bis-copper(II)-chloride complex. AB - A facile synthesis of a new bisoxazoline ligand is described. This ligand contains a urea bridging unit and is capable of stabilizing bimetallic complexes. An X-ray crystal structure of a bis-copper complex is reported. PMID- 20024212 TI - An organo-silver compound that shows antimicrobial activity against Pseudomonas aeruginosa as a monomer and plasma deposited film. AB - In this communication we describe the synthesis, characterisation and plasma deposition of a novel organo-silver compound for the prevention of the growth of Pseudomonas aeruginosa on both polystyrene surfaces and polypropylene non-woven fabrics. PMID- 20024213 TI - Hemoglobin bis-tetramers via cooperative azide-alkyne coupling. AB - Cross-linked hemoglobin-azides react with a bis-alkyne to form a bis-tetramer through sequential "click" reactions where the second step is promoted by the first. PMID- 20024214 TI - Fluoride-enhanced lanthanide luminescence and white-light emitting in multifunctional Al3Ln2 (Ln = Nd, Eu, Yb) heteropentanuclear complexes. AB - Multifunctional heteropentanuclear Al(3)Ln(2) (Ln = Nd, Eu, Yb) clusters with a novel structure exhibit significantly fluoride-enhanced lanthanide emission intensity and lifetime in both solid state and solution as well as unusual white light emitting for an Al(3)Eu(2) complex. PMID- 20024215 TI - Responsive and reactive terbium complexes with an azaxanthone sensitiser and one naphthyl group: applications in ratiometric oxygen sensing in vitro and in regioselective cell killing. AB - A terbium complex has been designed that may be used to sensitise formation of singlet oxygen (phi(em) 12%), allowing the creation of a ratiometric sensory system for pO(2) (versus the insensitive Eu analogue) and the definition of a reactive probe that can be used to cause local damage to cells loaded with the complex and exposed to laser excitation. PMID- 20024216 TI - Synthetic autocatalysts show organocatalysis of other reactions. AB - A molecule capable of both autocatalysis and organocatalysis was synthesized. The autocatalytic activity results from the self-complementary recognition sites provided by hydrogen bonding between heterocyclic subunits and the organocatalysis resides in an embedded thiourea function. The behavior of the molecule suggests both replication and metabolism can be engineered into synthetic compounds. PMID- 20024217 TI - Stereoselective gold-catalyzed cycloaddition of functionalized ketoenynes: synthesis of (+)-orientalol F. AB - A stereoselective gold-catalyzed [2 + 2 + 2] cycloaddition of ketoenynes substituted at the propargylic position with OR groups has been applied for the synthesis of (+)-orientalol F and pubinernoid B. PMID- 20024218 TI - Efficient palladium-catalyzed coupling reactions of aryl bromides and chlorides with phenols. AB - A convenient and general palladium-catalyzed coupling reaction of aryl bromides and chlorides with phenols was developed. Various functional groups such as nitriles, aldehydes, ketones and esters are well tolerated and the corresponding products are obtained in good to excellent yield. PMID- 20024219 TI - Low activation energy dehydrogenation of aqueous formic acid on platinum ruthenium-bismuth oxide at near ambient temperature and pressure. AB - Highly selective dehydrogenation of formic acid in water was observed at near ambient temperature on a metal/metal oxide catalyst composed of platinum ruthenium and bismuth with a low activation energy of 37.3 kJ mol(-1). PMID- 20024220 TI - Ni-catalyzed regioselective three-component coupling of alkyl halides, arylalkynes, or enynes with R-M (M = MgX', ZnX'). AB - A new method for the regioselective three-component cross-coupling of alkyl halides, alkynes, or enynes with organomagnesium or organozinc reagents in the presence of a nickel catalyst and a dppb ligand has been developed. PMID- 20024221 TI - Preparation of highly substituted tetrahydropyrans via a metal assisted dipolar cycloaddition reaction. AB - A range of highly substituted tetrahydropyrans have been prepared by reaction of a donor-acceptor cyclobutane, where the donor is a metal-alkyne complex, with an aldehyde under Lewis acid conditions. PMID- 20024222 TI - Aryloxide anions can form outer sphere complexes with metals as electropositive as uranium. AB - A uranium complex containing an outer sphere aryloxide anion is formed by the proteolytic cleavage of the methyl group in the mono-methyl uranium metallocene, (C(5)Me(5))(2)[(i)PrNC(Me)N(i)Pr]UMe, by 2,6-di-tert-butyl-4-methyl phenol. PMID- 20024223 TI - Chemical transformation and morphology change of nickel-silica hybrid nanostructures via nickel phyllosilicates. AB - Ni@SiO(2) core-shell nanoparticles were transformed to Ni particles on silica spheres via a branched nickel phyllosilicate phase by hydrothermal and hydrogen reduction reactions; the final morphology was successfully employed as an active nanocatalyst for the hydrogen transfer reaction of acetophenone. PMID- 20024224 TI - Multifunctional core-shell magnetic cisplatin nanocarriers. AB - Towards an integrated multifunctional nanocarrier, core-shell nanostructures have been developed using the electrostatic self-assembly of an organic shell onto magnetic nanoparticles. PMID- 20024225 TI - Solar light-driven photocatalyzed alkylations. Chemistry on the window ledge. AB - Tetrabutylammonium decatungstate photocatalysis is effective for the C-H activation of alkanes, aldehydes, ethers and amides and the alkylation of alkenes occurs effectively by merely exposing the solution to the sun in a glass vessel on a window ledge (up to 10 g per dm(2) in a few days). PMID- 20024226 TI - Hydrogen bonding-induced color recovery of gold nanoparticles upon conjugation of amino acids. AB - Hydrogen bonding-induced redispersion of the aggregated Au nanoparticles upon N hydroxysuccinimide ester bioconjugations may provide a simple and colorimetric tool as an optical sensor to detect a trace amount of amino acids as low as approximately 10(-6) M in an aqueous solution. PMID- 20024227 TI - Sensitive, selective and label-free protein detection using a smart polymeric transducer and aptamer/ligand system. AB - We have demonstrated a smart polymeric transducer and aptamer/intercalating dye system that allows the label-free detection of protein with high sensitivity and selectivity. PMID- 20024228 TI - Facile synthesis of carbon-decorated single-crystalline Fe3O4 nanowires and their application as high performance anode in lithium ion batteries. AB - A facile microwave-hydrothermal approach has been used to synthesize single crystalline Fe(3)O(4) nanowires within 15 min at 150 degrees C. The Fe(3)O(4) nanowires, after decorating with carbon, exhibit excellent cyclability and rate performance when employed as an anode in lithium ion batteries. PMID- 20024229 TI - A new approach to switching of enantioselectivity in NHC-Cu-catalyzed conjugate addition of alkylzincs to cyclic enones. AB - Conjugate addition of Et(2)Zn to 2-cyclohexen-1-one catalyzed by Cu(OTf)(2) combined with an azolium salt derived from (S)-leucinol produced the corresponding (S)-adduct, while the use of Cu(acac)(2) in combination with the same ligand afforded the (R)-adduct as a major product. PMID- 20024230 TI - Synthesis of fluorinated allenes via palladium-catalyzed monofluoromethylation using FBSM. AB - Palladium-catalyzed monofluoromethylation of substituted 2-bromo-1,3-dienes using fluorobis(phenylsulfonyl)methane (FBSM) as a pronucleophile gave previously unknown monofluoromethylated allenes in high yields, which are the isosteres of biologically attractive allenic alcohols. PMID- 20024231 TI - Highly enantio- and diastereoselective synthesis of alpha trifluoromethyldihydropyrans using a novel bifunctional piperazine-thiourea catalyst. AB - The first enantioselective Michael addition of alpha-cyanoketones to alpha,beta unsaturated trifluoromethyl ketones using a novel piperazine-thiourea catalyst was described. The resulting alpha-trifluoromethyldihydropyrans were obtained in high yields and with up to 95% ee within a short reaction time. A useful transformation of the chiral adduct was also illustrated. PMID- 20024232 TI - Facile synthesis of sulfonyl amidines via carbon-nitrogen bond formation mediated by FeCl3. AB - A facile synthesis of sulfonyl amidines via carbon-nitrogen bond formation mediated by FeCl(3) was developed and an interesting major product of cyclic tertiary amine was observed, which showed the good selectivity of FeCl(3) mediated activation of cyclic alpha-C-H bonds of cyclic tertiary amines. PMID- 20024233 TI - Probing the structural factors influencing columnar mesophase formation and stability in triphenylene discotics. AB - Series of structurally related substituted triphenylene derivatives were designed and synthesised to interrogate key features which determine mesophase formation and stability, and to challenge the general conclusions previously proposed by us and others. It is apparent that no single, simple principle can be universally applied. PMID- 20024234 TI - Synthesis and validation of a probe to identify quorum sensing receptors. AB - The synthesis and evaluation of a 'tag-free' probe to isolate and identify receptors for N-acyl homoserine lactones is described. PMID- 20024235 TI - Thermal rearrangement of thiocarbonyl-stabilised triphenylphosphonium ylides leading to (Z)-1-diphenylphosphino-2-phenylsulfenylalkenes. AB - While thermolysis of thiocarbonyl-stabilised phosphonium ylides generally results in extrusion of Ph(3)PS to give alkynes, those with a P=CH function instead undergo a novel P to S transfer of a phenyl group to give (Z)-configured 1 phosphino-2-sulfenylalkenes of interest as bidentate ligands. PMID- 20024236 TI - A reversible fluorescent Hg2+/K+ switch that works as keypad lock in the presence of F- ion. AB - A thiacalix[4]arene based fluorescent chemosensor 3 of 1,3-alternate conformation bearing two dansyl groups and a crown-5 ring behaves as an "On-Off" reversible switch for two chemical inputs Hg(2+) and K(+) ions and mimics a molecular level keypad lock in the presence of F(-) ions. PMID- 20024237 TI - Conjugated ligand-based tribochromic luminescence. AB - A Au(I) complex containing a terthienyl diphosphine ligand is non-emissive in the crystalline form, but exhibits intense ligand-based emission upon grinding attributed to increased planarization in the terthienyl ligand. PMID- 20024238 TI - Sulfide-selective chemosignaling by a Cu2+ complex of dipicolylamine appended fluorescein. AB - A new sulfide-selective chemosignaling system was devised based on a Cu(2+) complex of fluorescein derivative having a dipicolylamine (DPA) binding site; the fluorescein-DPA conjugate 1, in the presence of Cu(2+) ions, revealed a selective turn-on type signaling behavior toward sulfide ions with a detection limit of 420 nM in 100% aqueous solution. PMID- 20024239 TI - Steric effects direct the binding of porphyrins to tetramolecular quadruplex DNA. AB - The binding motifs of copper(II) porphyrins with G quadruplex DNA structures vary markedly depending on the steric demands of the ligand and the host structure. PMID- 20024240 TI - Enhanced reversible electrochromism via in situ phase transformation in tungstate monohydrate. AB - This study demonstrates that realizing the correlation between in situ crystallographic structure modifications of an electrochromic material and its functionality leads to improved performances, which can then contribute to a variety of energy-efficient applications. PMID- 20024241 TI - Products of the iterative polyketide synthases in 9- and 10-membered enediyne biosynthesis. AB - The iterative polyketide synthases from the biosynthetic pathways of three enediyne natural products were examined. The results established the all-trans conjugated pentadecaheptanene as the only major product shared by the PKSs. The experiments further revealed some intrinsic differences among the PKSs by demonstrating the formation of different by-products. PMID- 20024242 TI - Heterogeneity in the stereochemistry of mycolactones isolated from M. marinum: toxins produced by fresh vs. saltwater fish pathogens. AB - A novel mycolactone has been identified from Mycobacterium marinum infecting freshwater fish. PMID- 20024243 TI - A new class of double alkyl-substituted, liquid crystalline imidazolium ionic liquids--a unique combination of structural features, viscosity effects, and thermal properties. AB - 1,3-Didodecylimidazolium salts have been prepared as first representatives of a new class of ionic liquid crystals (ILCs), showing thermotropic liquid crystalline behaviour in an extended temperature range below 70 degrees C. Remarkably strong non-Newtonian viscosity behaviour was found for the liquid crystalline state of these ILCs. PMID- 20024244 TI - A fluorescent heteroditopic ligand responding to free zinc ion over six orders of magnitude concentration range. AB - A fluorescent heteroditopic ligand useful in live-cell imaging studies responds to free zinc ion concentration over a range of six orders of magnitude in a buffered aqueous solution via dual-channel fluorescence. PMID- 20024245 TI - Preparation of organic nanoscrews from simple porphyrin derivatives. AB - Twisted supramolecular assemblies "nanoscrews", were prepared by using simple porphyrin derivatives and acetonitrile solvent. The electrical properties of the assemblies were measured by using microgap electrodes. The nanoscrew had conductivity and showed photo-current. By changing the solvent, the pitch of the screws and their aggregation shapes could be controlled. PMID- 20024246 TI - Facile syntheses of bimetallic ytterbium bisamides stabilized by a flexible bridged bis(phenolato) ligand and the high activity for the polymerization of L lactide. AB - The "ate" and neutral bimetallic ytterbium bisamido complexes stabilized by a flexible bridged bis(phenolate) group are prepared, which exhibited high activity for the ring-opening polymerization of L-lactide. PMID- 20024247 TI - 1,2-Olefin addition of a frustrated amine-borane Lewis pair. AB - Addition of B(C(6)F(5))(3) to an alpha-dimethylamino-o-vinylferrocenophane system gives a frustrated Lewis pair that undergoes intramolecular 1,2-N-B addition to the alkene. PMID- 20024248 TI - Label-free dual assay of DNA sequences and potassium ions using an aptamer probe and a molecular light switch complex. AB - A homogeneous assay is reported using Ru(phen)(2)(dppz)(2+) and a K(+)-binding aptamer for the selective and sensitive detection of a target oligonucleotide and potassium ions, based on reduction in fluorescence emission according to the formation of the G-quadruplex structure from the aptamer in the presence of K(+). PMID- 20024249 TI - Two-photon absorption properties of hexa-substituted benzene derivatives. Comparison between dipolar and octupolar molecules. AB - Hexa-substituted benzene derivatives show larger two-photon cross sections than the corresponding dipolar counterparts and the largest values of delta(max) = 2000 GM and delta(max)/MW = 2.25 GM g(-1) have been obtained with a compound having C[triple bond]C and C=C bonds in the conjugation bridge. PMID- 20024250 TI - Mechanism of enantioselection in Rh-catalyzed asymmetric hydrogenation. The origin of utmost catalytic performance. AB - This article describes recent developments in the understanding of the mechanism of enantioselection in one of the most efficient artificial catalytic reactions- Rh-catalyzed asymmetric hydrogenation. Numerous experimental and computational data are consistent with the idea that the enantioselection takes place through the reversible coordination of the double bond in octahedral dihydride complexes, which can happen only if the resulting chelate cycle is formed in the less hindered quadrant. In its main features, this mechanism of generating chirality resembles the recently uncovered three-stage recognition process occurring in enzymatic reactions. PMID- 20024251 TI - Selective difluoromethylation and monofluoromethylation reactions. AB - The selective introduction of fluorine atom(s) and fluorinated moieties into organic molecules has become an important and fast-growing research field, since fluorine atoms play crucial roles in life science and materials science-related applications. Similar to the trifluoromethyl group, both difluoromethyl and monofluoromethyl groups can often bring about many beneficial effects to the target molecules, and a variety of CF(2)H- and CH(2)F-containing pharmaceuticals and agrochemicals have been developed. Among the synthetic methods for CF(2)H- and CH(2)F-containing compounds, selective di- and monofluoromethylation (i.e., introduction of CF(2)H and CH(2)F groups into organic molecules) represent one of the most straightforward synthetic methods and thus can be conveniently used in the synthetic design. This feature article summarizes the presently known selective difluoromethylation and monofluoromethylation methods, including nucleophilic, electrophilic, and free radical di- and monofluoromethylation reagents and reactions. PMID- 20024252 TI - A selective oligonucleotide-based luminescent switch-on probe for the detection of nanomolar mercury(II) ion in aqueous solution. AB - An oligonucleotide-based luminescent platinum(II) switch-on probe has been developed for selective detection of nanomolar Hg(2+) ions. PMID- 20024253 TI - Selectivity in small molecule binding to human telomeric RNA and DNA quadruplexes. AB - Quadruplex RNAs are less well understood than their DNA counterparts, yet of potentially high biological relevance. The interactions of several quadruplex binding ligands with telomeric RNA quadruplexes are reported and compared with their binding to the analogous DNA quadruplexes. PMID- 20024254 TI - Emission quench of water-soluble ZnS-AgInS2 solid solution nanocrystals and its application to chemosensors. AB - Redox-dependent emission quenching of low toxic ZnS-AgInS(2) semiconductor nanocrystals was studied and the obtained behavior was found to be applicable to fabrication of fluorescent biosensors in combination with redox enzymes. PMID- 20024255 TI - Quinone tailored selective oxidation of methane over palladium catalyst with molecular oxygen as an oxidant. AB - With the in situ generated H(2)O(2) tailored by the addition of p tetrachlorobenzoquinone, the product can be effectively steered towards either HCOOH or the methanol derivative CF(3)COOCH(3) during the direct oxidation of methane with molecular oxygen over palladium catalyst. PMID- 20024256 TI - Vectorial growth/regulations in a {P8W48}-type polyoxotungstate compartment: trapped unusual molybdenum oxide acts as a handle. AB - Reaction of the cyclic {P(8)W(48)} polyoxotungstate host with sodium molybdate in solution in the presence of a reducing agent leads to the formation and stabilization of unprecedented neutral {Mo(V)(4)O(10)(H(2)O)(3)} aggregates with handle function, thereby proving the potential of the present host for performing future interesting studies related to mixed-valence type chemistry under confined conditions. PMID- 20024257 TI - Highly ordered transparent mesoporous TiO2 thin films: an attractive matrix for efficient immobilization and spectroelectrochemical characterization of cytochrome c. AB - We demonstrate remarkably fast incorporation and high loading of cytochrome c within thin films of periodically ordered nanocrystalline TiO(2) deposited on transparent electrodes. The immobilized cytochrome c is not denaturated and it can be reversibly reduced without mediator over the time scale of a few seconds as evidenced by spectroelectrochemistry. PMID- 20024258 TI - Self-assembled, cogged hexameric nanotubes formed from pyrogallol[4]arenes with a unique branched side chain. AB - A branched side chain pyrogallol[4]arene self-assembles into a previously unreported cogged hydrogen-bonded nanotube structure in the solid state and self assembles at the air-water interface on the Langmuir trough. PMID- 20024259 TI - Switching and tuning organic solid-state luminescence via a supramolecular approach. AB - Unusual intermolecular interactions of organic luminescent acid, 2-cyano-3(4 (diphenylamino)phenyl)acrylic acid (CDPA), with amines lead to the formation of supramolecular luminescence systems with switchable and tunable solid-state luminescence. PMID- 20024260 TI - Affinity ionic liquid. AB - An affinity ionic liquid, based on biomolecular recognition, was developed and found to be capable of quantitative partitioning of biomacromolecules from aqueous buffer to ionic liquid. PMID- 20024261 TI - Two distinct beta-sheet fibrils from silk protein. AB - This study showed that Bombyx mori silk protein could be selectively induced to fold into fibrils dominated by either cross- or parallel-beta-sheet structure, where the beta-strands arrange perpendicular or parallel to the long fibril axis, incubated in ethanol-water quiescently or in water under shear. PMID- 20024262 TI - Laser printing mediated cell patterning. AB - An approach for complex cell patterning, using laser printing, is described allowing essentially any cellular image or pattern to be rapidly fabricated. PMID- 20024263 TI - The commensurate modulated structure of the metastable state in spin crossover complex [Fe(abpt)2(NCS)2]. AB - Two new polymorphs (C and D) of [t-Fe(abpt)(2)(NCS)(2)] are characterized, a commensurate modulated structure with the c axis tripled in the thermally quenched metastable high spin state at 25 K is found in polymorph C. PMID- 20024264 TI - Glutathione-bound gold nanoclusters for selective-binding and detection of glutathione S-transferase-fusion proteins from cell lysates. AB - A straightforward method for the rapid detection of the presence of glutathione S transferase (GST)-tagged proteins from sample solutions using glutathione (GSH) bound gold nanoclusters (Au@GSH NCs) with luminescence properties as the detection probes by simple observation with the naked eye was proposed in this study. PMID- 20024265 TI - Ultrathin tough double network hydrogels showing adjustable muscle-like isometric force generation triggered by solvent. AB - Ultrathin double-network hydrogels, which have super-high toughness under micro scale thickness (elastic elongation epsilon(b) > 1000%, tensile strength sigma(b) > 2 MPa and tearing energy G approximately 600 J m(-2)), and solvent-triggered fast and high isometric stress generation, were synthesized by coupling the salt controlled swelling process and polymer chain pre-reinforced technique. PMID- 20024266 TI - Radiolabelling of proteins with fluorine-18 via click chemistry. AB - The study describes for the first time the application of Cu(I)-mediated 1,3 dipolar [3+2]cycloaddition for the labelling of proteins with the short-lived positron emitter fluorine-18 as exemplified with azide-functionalized human serum albumin (HSA). PMID- 20024267 TI - Mechanism of supported gold nanoparticles as photocatalysts under ultraviolet and visible light irradiation. AB - Gold nanoparticles strongly absorb both visible light and ultraviolet light to drive an oxidation reaction for a synthetic dye, as well as phenol degradation and selective oxidation of benzyl alcohol under UV light. PMID- 20024268 TI - In situ neutron powder diffraction and structure determination in controlled humidities. AB - A controlled-humidity sample environment has been constructed, allowing bulk powder samples undergoing humidity-induced phase transitions and reactions to be studied via in situ neutron diffraction. Associated developments in data collection and analysis permit this to be achieved without the use of D(2)O. PMID- 20024269 TI - Towards black chromophores: mu-oxo linked phthalocyanine-porphyrin dyads and phthalocyanine-subphthalocyanine dyad and triad arrays. AB - A versatile synthetic strategy based on the use of a non-peripherally substituted dihydroxysilicon phthalocyanine and dihydroxygermanium octaethylporphyrin has permitted controlled construction of heterodyads that absorb across the whole visible spectrum; the same phthalocyanine also reacts with chlorosubphthalocyanine to form a Si-O-B linked dyad and triad. PMID- 20024270 TI - Template synthesis of nanoparticle arrays of CdS in transparent layered double hydroxide films. AB - Highly ordered transparent composite films of CdS and layered double hydroxides (LDHs) have been fabricated using a template synthesis method; the size of the CdS particles in the LDH host can be controlled by adjusting the length of time of the reaction with H(2)S. PMID- 20024271 TI - Enhanced lithium storage capacity and cyclic performance of nanostructured TiO2 MoO3 hybrid electrode. AB - New porous heterostructure of TiO(2)-MoO(3) nanohybrids have been successfully synthesized via an exfoliation and reassembling method: the hybridization between TiO(2) and MoO(3) gives rise to a remarkable enhancement of the reversible capacity of 420 mAhg(-1). PMID- 20024272 TI - A new open benzodipyrrole-based chemosensor for hydrogenpyrophosphate anion in aqueous environment. AB - Benzodipyrrolic derivatives have been synthesised and evaluated as neutral receptors for anions, in a mixed acetone-water media. PMID- 20024273 TI - Facile synthesis of nickel oxide nanotubes and their antibacterial, electrochemical and magnetic properties. AB - Nickel oxide nanotubes with great antibacterial activities, electrochemical capacitance, and magnetic properties have been synthesized through a precursor method with dimethylglyoxime as precipitant for the precursor, and the method has been developed for the synthesis of Ni/C nanorods. PMID- 20024274 TI - Enhanced electrochemiluminescence efficiency of Ru(II) derivative covalently linked carbon nanotubes hybrid. AB - The synthesized derivative Ru(bpy)(3) covalently linked CNTs hybrid shows good electrochemical activity and ca. 17 times higher luminescence quantum efficiency than the adsorbed derivative Ru(bpy)(3). The Ru-CNTs based ECL sensor exhibits high stability toward determination of TPA with a detection limit as low as 8.75 pM. PMID- 20024275 TI - Fluorescent mimics of 5-hydroxytryptamine based on N-alkylated derivatives of 6 hydroxycarbostyril. AB - Fluorescent probes based on a 6-hydroxycarbostyril core accumulate inside neurons and astroglia in the absence of a serotonin uptake inhibitor. PMID- 20024276 TI - A new MOF-505 analog exhibiting high acetylene storage. AB - A new microporous metal-organic framework Cu(2)(EBTC)(H(2)O)(2) x xG (EBTC = 1,1' ethynebenzene-3,3',5,5'-tetracarboxylate; G = guest molecule) was rationally designed with a NbO net, exhibiting significantly high acetylene storage of 252 and 160 cm(3) g(-1) at 273 and 295 K under 1 bar, respectively. PMID- 20024277 TI - Self-assembly of cerium-based metal-organic tetrahedrons for size-selectively luminescent sensing natural saccharides. AB - New Ce-based Werner type tetrahedrons were achieved for size-selectively luminescent detection of natural carbohydrates through incorporating amide groups as both the multiple hydrogen bonding triggers and binding-signalling transductor. PMID- 20024278 TI - Ceria-terbia solid solution nanobelts with high catalytic activities for CO oxidation. AB - Ceria-terbia solid solution nanobelts were prepared by an electrochemical route and tested as catalysts of high activity for CO oxidation. PMID- 20024279 TI - Anisotropic ionic conductivities in lyotropic supramolecular liquid crystals. AB - The designed aromatic amide discotic molecule with sulfonic acid groups at its periphery exhibits a hexagonal supramolecular columnar liquid crystalline phase, which leads to the achievement of anisotropic ionic conductivity through macroscopically aligning the ionic channels. PMID- 20024280 TI - Transformation of framework solids into processable metallo-polymers. AB - A novel design strategy that extends metal-organic frameworks (MOFs) into tractable metallo-polymers was successfully explored by incorporating a homologous series of alkyl side chains to an organic linker, in which drastic solid to solid transformation is also observed. PMID- 20024281 TI - Synthesis of transition metal isocyanide compounds from carbonyl complexes via reaction with Li[Me3SiNR]. AB - The reaction between a transition metal carbonyl compound, L(n)MCO, and Li[Me(3)SiNR] yields the corresponding isocyanide derivative, L(n)MCNR, thereby providing a new route to transition metal isocyanide compounds that does not require the use of free isocyanides as reagents. PMID- 20024282 TI - Controlled synthesis of Co3O4 nanopolyhedrons and nanosheets at low temperature. AB - Cobalt oxide (Co(3)O(4)) nanopolyhedrons and nanosheets were controlled synthesized at the low temperature of 80 degrees C via a novel, simple, aqueous method; the obtained nanosheets are only 2-3 nm thick. PMID- 20024283 TI - Catalytic production of sulfur heterocycles (dihydrobenzodithiins): a new application of ligand-based alkene reactivity. AB - Activation of bis-o-phenylene tetrasulfide to render it a practical benzodithiete equivalent for [4+2] cycloadditions with alkenes has been achieved with catalytic amounts of Mo(tfd)(2)(bdt) (tfd = S(2)C(2)(CF(3))(2); bdt = S(2)C(6)H(4)). Substituted 2,3-dihydro-1,4-benzodithiins are produced. PMID- 20024284 TI - Mesoporous nanowire array architecture of manganese dioxide for electrochemical capacitor applications. AB - Mesoporous MnO(2) nanowire array architecture exhibits enhanced capacitive and charge/discharge performance for electrochemical capacitors. PMID- 20024285 TI - Direct fabrication of mesoporous zeolite with a hollow capsular structure. AB - We report the direct fabrication of mesoporous zeolite with a hollow spherical/ellipsoidal capsule structure by using conventional TPAOH and CTAB as soft micro- and mesopore generating templates; the mesoporous zeolite shows a high surface area of 717 m(2) g(-1) and small mesopore size of around 3 nm. PMID- 20024287 TI - Surfactant-free synthesis of mesoporous and hollow silica nanoparticles with an inorganic template. AB - A surfactant-free synthesis of mesoporous and hollow silica nanoparticles is reported in which boron acts as the templating agent. Using such a simple and mild procedure as a treatment with water, the boron-rich phase is selectively removed, affording mesoporous pure silica nanoparticles with wormhole-like pores or, depending on the synthetic conditions, silica nanoshells. PMID- 20024286 TI - Assembly of indole-2-carboxylic acid esters through a ligand-free copper catalysed cascade process. AB - A straightforward synthesis of indole-2-carboxylic esters was developed through a ligand-free copper-catalysed condensation/coupling/deformylation cascade process from 2-halo aryl aldehydes or ketones with ethyl isocyanoacetate. The reactions proceeded well for most of the 2-iodo-, bromo-, and chloro-substrates under room temperature or mild conditions. PMID- 20024288 TI - Palladium(II)-catalyzed coupling reactions with a chelating vinyl ether and arylboronic acids: a new Heck/Suzuki domino diarylation reaction. AB - A mild and novel palladium(II)-catalyzed domino Heck/Suzuki alpha,beta diarylation-reduction of a dimethylaminoethyl substituted chelating vinyl ether was developed by using electron-rich arylboronic acids in combination with p benzoquinone. Based on the preparative results, a catalytic cycle is proposed. Further, highly regioselective palladium(II)-catalyzed alpha- or beta monoarylation of the chelating vinyl ether was achieved using either a bidentate ligand or by employing ligand-less conditions. PMID- 20024289 TI - A hierarchical Fe/ZSM-5 zeolite with superior catalytic performance for benzene hydroxylation to phenol. AB - We report the one-step synthesis of a highly active hierarchical Fe/ZSM-5 zeolite catalyst with a strongly improved lifetime in the selective hydroxylation of benzene to phenol with nitrous oxide; compared to the best Fe/ZSM-5 catalyst, the turnover number after 24 h on stream is almost four times higher. PMID- 20024291 TI - Advances in chemical ligation strategies for the synthesis of glycopeptides and glycoproteins. AB - A number of recent advances in the chemical synthesis of glycopeptides and glycoproteins are described, with particular focus on the development of peptide ligation strategies and their implementation in the convergent assembly of complex glycopeptides. Recent applications in the synthesis of full length homogeneous glycoproteins are also highlighted. PMID- 20024292 TI - Gas storage in porous metal-organic frameworks for clean energy applications. AB - Depletion of fossil oil deposits and the escalating threat of global warming have put clean energy research, which includes the search for clean energy carriers such as hydrogen and methane as well as the reduction of carbon dioxide emissions, on the urgent agenda. A significant technical challenge has been recognized as the development of a viable method to efficiently trap hydrogen, methane and carbon dioxide gas molecules in a confined space for various applications. This issue can be addressed by employing highly porous materials as storage media, and porous metal-organic frameworks (MOFs) which have exceptionally high surface areas as well as chemically-tunable structures are playing an unusual role in this respect. In this feature article we provide an overview of the current status of clean energy applications of porous MOFs, including hydrogen storage, methane storage and carbon dioxide capture. PMID- 20024293 TI - Mechanically interlocked and switchable molecules at surfaces. AB - Mechanically interlocked molecules are of considerable interest from data storage, molecular scale or sensory device perspectives. Though the solution phase characterisation of these compounds has been extensively explored, progress towards real world application will, in many cases, necessitate a detailed understanding of their interfacing with supportive, optically transparent or electroactive surfaces. This feature article summarises the developments made in surface assembly and characterisation including recent progress in exploiting templating methods to interlock molecular systems on surfaces. PMID- 20024294 TI - Reductive dissolution of Fe3O4 facilitated by the Au domain of an Fe3O4/Au hybrid nanocrystal: formation of a nanorattle structure composed of a hollow porous silica nanoshell and entrapped Au nanocrystal. AB - The Fe(3)O(4) grain of a Fe(3)O(4)/Au hybrid nanocrystal encapsulated in a silica nanosphere was rapidly and exclusively dissolved through a reductive process facilitated by the attached Au grain, resulting in the formation of a nanorattle structure which has utility as a nanoreactor to template the growth of nanocrystals inside the cavity. PMID- 20024295 TI - Self-association based on orthogonal C=O...C=O interactions in the solid and liquid state. AB - A network of orthogonal C=O...C=O interactions was identified in the X-ray crystal structure of an alpha,alpha-difluorocyclopentanone derivative. This finding inspired investigations of self-association driven by these weak dipolar interactions in apolar solvents, which was proven by (1)H NMR spectroscopy. PMID- 20024296 TI - Synthesis of functionalized maoecrystal V core structures. AB - Two strategies toward the total synthesis of maoecrystal V (1) culminating in the construction of core structures 2 and 3 are described. PMID- 20024297 TI - Highly dispersible, superparamagnetic magnetite nanoflowers for magnetic resonance imaging. AB - A one-pot reaction process was developed to synthesize highly dispersible, superparamagnetic Fe(3)O(4) nanoflowers; the potential of these nanoflowers as MRI contrast agents was investigated. PMID- 20024299 TI - Decarbonylation of ethanol to methane, carbon monoxide and hydrogen by a [PNP]Ir complex. AB - The putative three-coordinate Ir(i) PNP(Pr(i)) (PNP(Pr(i)) = [N{2-P(CHMe(2))(2)-4 MeC(6)H(3)}(2)](-)) pincer complex decarbonylates ethanol to yield methane, hydrogen and [PNP(Pr(i))]Ir(CO). The mechanism involves the isolable trans [PNP(Pr(i))]Ir(H)(Me)(CO), which is susceptible to photochemical reductive elimination of methane. PMID- 20024298 TI - Bioluminescent nanosensors for protease detection based upon gold nanoparticle luciferase conjugates. AB - This communication reports the use of click chemistry to site-specifically conjugate bioluminescent Renilla luciferase proteins to gold nanoparticles (Au NPs) for sensing protease activity. The bioluminescent emission from luciferase was efficiently quenched by Au NPs, but significantly recovered after the proteolytic cleavage. PMID- 20024300 TI - Conformational polymorphism facilitates assignment of trans and cis-conformers of an alpha-substituted oligothiophene via IR spectroscopy. AB - Conformational polymorphism is exploited as a means to assign cis-trans and trans trans conformations of an oligothiophene backbone using solid-state IR spectroscopy. PMID- 20024301 TI - Single and double substrate insertion into the Ti=N(alpha) bonds of terminal titanium hydrazides. AB - Nitriles, CO(2) and isocyanates undergo net single or double insertion reactions into the Ti=N(alpha) multiple bonds of terminal titanium hydrazides. These are the first such examples of this type of reactivity for any transition metal hydrazide complex. PMID- 20024302 TI - Formation of tetrameric water clusters driven by a cavitand template. AB - We describe an innovative approach to the generation of tetrameric water clusters in the solid state. The specific H-bond pattern induced by the tetraphosphonate cavitand template via its rigidly preorganized P=O acceptor groups leads to the exclusive formation of the unique cyclic homodromic water tetramer of C(4) symmetry. PMID- 20024303 TI - Thioether S-ligation in a side-on micro-eta2:eta2-peroxodicopperii complex. AB - [(ANS)Cu(I)(CH(3)CN)](+) reacts with O(2) giving [{(ANS)Cu(II)}(2)(micro eta(2):eta(2)-O(2)(2-))](2+), nu(O-O) = 731 cm(-1), shown to possess S-thioether ligation, based on comparisons with analogues having all N-ligands or a -S(Ph) group. The finding is a rare occurrence and new for side-on O(2)(2-) binding. PMID- 20024304 TI - Enzyme logic gate associated with a single responsive microparticle: scaling biocomputing to microsize systems. AB - A microsize biocomputing system based on enzyme logic processing biochemical signals was developed. Optical transduction of pH signals generated in situ by the enzyme OR logic gate was achieved with the use of a single optode microparticle. PMID- 20024305 TI - Aza-dibenzocyclooctynes for fast and efficient enzyme PEGylation via copper-free (3+2) cycloaddition. AB - A strained aza-dibenzocyclooctyne was prepared via a high-yielding synthetic route. Copper-free, strain-promoted click reaction with azides showed excellent kinetics, and a functionalised aza-cyclooctyne was applied in fast and efficient PEGylation of enzymes. PMID- 20024306 TI - Subtleties in asymmetric catalyst structure: the resolution of a 6-phospha-2,4,8 trioxa-adamantane and its applications in asymmetric hydrogenation catalysis. AB - An efficient, classical resolution of the versatile P-ligand intermediate 6 phospha-2,4,8-trioxa-adamantane (CgPH) is described and the rhodium complex of the optically pure secondary phosphine beta-CgPH is an active and moderately selective asymmetric hydrogenation catalyst. PMID- 20024307 TI - Multimodal cell imaging by ruthenium polypyridyl labelled cell penetrating peptides. AB - The capacity of ruthenium polypyridyl complexes as probes for combined confocal luminescence and resonance Raman imaging, enabled by their large Stokes shift, is demonstrated for a novel membrane sensitive Ru(ii) polypyridyl peptide. Confocal luminescence and resonance Raman imaging provides complementary information about the membrane phospholipid regions of the cell and the location of the dye within the cell. PMID- 20024309 TI - ssPNA templated assembly of oligo(p-phenylenevinylene)s. AB - A single-stranded oligothymine peptide nucleic acid (PNA) was used as a template for the assembly of a chiral oligo(p-phenylenevinylene) diaminotriazine derivative (OPV) in methylcyclohexane (MCH) revealing nanostructures in which the size is controlled by the template. PMID- 20024308 TI - Protein nanopatterning and release from gold nano-cavity arrays. AB - Selective chemical modification of a gold nano-cavity array is achieved via nanoscale templating to create fibrinogen patterned cavities with a polyethylene glycol modified top surface. Application of a reducing potential to the array readily releases the protein from the cavities. PMID- 20024310 TI - Molecular tectonics: tubular crystals with controllable channel size and orientation. AB - The combination of flexible neutral organic tectons based on two pyridines interconnected by a thioether or thioester type spacer with an inorganic ZnSiF(6) pillar leads to the formation of 2-D coordination networks and the packing of the latter generates crystals offering controllable tubular channels with imposed orientation along the pillar axis. PMID- 20024311 TI - Molecular tectonics: from 1-D interwoven racemic chains to quadruple-stranded helices. AB - The combination of two positional isomers tectons 1 and 2, based on a racemic 1,1'-spirobi(indane) scaffold bearing two pyridine units, with HgCl(2) affords doubly interwoven and quadruple-stranded helical architectures, respectively. PMID- 20024312 TI - Facile scalable synthesis of magnetite nanocrystals embedded in carbon matrix as superior anode materials for lithium-ion batteries. AB - A simple and scalable process was developed for the synthesis of highly crystalline magnetite nanocrystals embedded in a carbon matrix using low cost starting materials; the resulting nanocomposite showed a very high specific capacity of 863 mA hg(-1) in the initial cycle and high capacity retention of 90% after 30 cycles. PMID- 20024314 TI - Hydrophobic chromophore cargo in micellar structures: a different strategy to sensitize lanthanide cations. AB - We propose a new approach for the versatile sensitization of luminescent lanthanide cations. A hydrophobic chromophore is incorporated into a micellar assembly formed by the amphiphilic lanthanide chelate. The sensitizer to lanthanide energy transfer occurs between the two moieties without covalent linkage. PMID- 20024315 TI - Synthesis and photophysical properties of a hydrogen-bonded phthalocyanine perylenediimide assembly. AB - A supramolecular phthalocyanine-perylenediimide donor-acceptor array has been assembled by using a melamine/perylenediimide motif. Photoexcitation of the perylenediimide component affords transduction of singlet excited state energy to the energetically lower lying phthalocyanine. PMID- 20024313 TI - Shell crosslinked nanoparticles carrying silver antimicrobials as therapeutics. AB - Amphiphilic polymer nanoparticles loaded with silver cations or/and N heterocyclic carbene-silver complexes were assessed as antimicrobial agents against Gram-negative pathogens Escherichia coli and Pseudomonas aeruginosa. PMID- 20024316 TI - Synthesis of 3-hydroxyoxindoles by Pd-catalysed intramolecular nucleophilic addition of aryl halides to alpha-ketoamides. AB - Pd/PtBu(3)-catalysed intramolecular nucleophilic addition of aryl halides to alpha-ketoamides in the presence of nBuOH and base has been realized with high yields, providing a new, direct, and efficient synthetic strategy to obtain 3 hydroxyoxindoles. PMID- 20024317 TI - Dioxygen mediated hydroacylation of vinyl sulfonates and sulfones on water. AB - Herein we report a mild, facile method for the preparation of 1,4-keto-sulfonates and sulfones on water. Further synthetic manipulations can result in products that are not readily accessed by hydroacylation of electron rich alkenes. PMID- 20024318 TI - A short, unsupported CuI...CuI interaction, 2.65 A, in a dinuclear guanidine chloride complex. AB - The reaction of CuCl with the neutral guanidine Hhpp ligand (1,3,4,6,7,8 hexahydropyrimido[1,2-a]pyrimidine) yields two crystalline polymorphs of the neutral dimer, bis(1,3,4,6,7,8-hexahydropyrimido[1,2-a]pyrimidine) dichloro-di copper(I) with the shortest distance known for a bridging unsupported copper(I) copper(I) interaction. PMID- 20024319 TI - An enzymatic route to 5-deoxy-5-[18F]fluoro-D-ribose, a [18F]-fluorinated sugar for PET imaging. AB - An efficient two-step, one-pot, biotransformation involving the fluorinase enzyme is described for the synthesis of 5-deoxy-5-[(18)F]fluororibose, a novel [(18)F] fluorinated sugar suitable for positron emission tomography (PET) applications. PMID- 20024320 TI - Polynitramino compounds outperform PETN. AB - New polynitramino compounds were synthesized and fully characterized using IR and multinuclear ((1)H, (13)C, (15)N) NMR spectroscopy, and elemental analysis as well as single-crystal X-ray diffraction. PMID- 20024321 TI - Enhanced acyl radical formation in the Au nanoparticle-catalysed aldehyde oxidation. AB - EPR spectroscopy and spin-trapping experiments showed that polymer-encapsulated Au nanoparticles promote aldehyde oxidation via a radical pathway by initiating formation of acyl radicals. PMID- 20024322 TI - Recycle of tin thiolate compounds relevant to ammonia-borane regeneration. AB - The use of benzenedithiol as a digestant for ammonia-borane spent fuel has been shown to result in tin thiolate compounds which we demonstrate can be recycled, yielding Bu(3)SnH and ortho-benzenedithiol for reintroduction to the ammonia borane regeneration scheme. PMID- 20024323 TI - Pd-catalyzed cascade carbopalladation-annulation reaction of 3-(2-iodobenzyl) indoles into fused 6/5/7/6- and 6/5/5/6- heterocyclic systems. AB - Polycyclic indole structures, possessing fused seven-membered rings were efficiently synthesized via the Pd-catalyzed intramolecular carbopalladation annulation of 3-(2-iodobenzyl)-indoles and alkynes. A remarkable base effect on the chemoselectivity of this transformation has been found: switching from Et(3)N to CsOAc completely reverses the reaction path to intramolecular cyclization forming fused five-membered rings. PMID- 20024324 TI - Total synthesis of largamide H. AB - Total synthesis of largamide H has been completed, utilising the oxidative elimination reaction of enantiomerically pure 2-amino-3-(phenylselenyl)butanoic acid residues to stereospecifically install both (Z)- and (E)-2,3-dehydro-2 aminobutanoic moieties. PMID- 20024325 TI - Highly enantioselective hydrogenation of alpha-aryl-beta-substituted acrylic acids catalyzed by Ir-SpinPHOX. AB - The enantioselective hydrogenation of a series of challenging substrates, alpha aryl-beta-substituted acrylic acids, was realized with high efficiency and enantioselectivity (up to 96%) under the catalysis of Ir(i) complex of Spiro based P,N ligand, SpinPHOX. PMID- 20024326 TI - Ortho-C-H borylation of benzoate esters with bis(pinacolato)diboron catalyzed by iridium-phosphine complexes. AB - Iridium complexes generated from [Ir(OMe)(COD)](2) and tris[3,5 bis(trifluoromethyl)phenyl]phosphine efficiently catalyzed the ortho-C-H borylation of benzoate esters with bis(pinacolato)diboron in octane at 80 degrees C to produce the corresponding arylboronates in high yields with excellent regioselectivities. PMID- 20024327 TI - Application of d6 transition metal complexes in fluorescence cell imaging. AB - Luminescent d(6) transition metal complexes have often been proposed as useful fluorophores for cell imaging due to their attractive photophysical attributes, but until very recently their actual applications have been scarce, and largely limited to ruthenium complexes in DNA and oxygen sensing. In the last few years, however, there has been an increasing number of reports of the design and application in cellular studies of a diverse range of Ir, Re and Ru complexes tailor-made for imaging applications. The design principles, uptake and cellular localisation of this new class of imaging agents are presented in context in this feature article. PMID- 20024328 TI - Bis(oxazolinyl)phenyl transition-metal complexes: asymmetric catalysis and some reactions of the metals. AB - Several transition-metal complexes that bind the bis(oxazolinyl)phenyl moiety as a tridentate ligand have been prepared to showcase their highly potent activities for asymmetric catalysis in conjugate reductions, reductive aldol reactions, direct aldol reactions and hydrogenation reactions. Some reactions of the metals were also examined. PMID- 20024329 TI - Silver(I)-mediated highly enantioselective synthesis of axially chiral allenes under thermal and microwave-assisted conditions. AB - Silver(I) salts mediated stereospecific transformation of optically active propargylamines to axially chiral allenes with excellent enantioselectivities (17 examples with 96-99% ee; one substrate with 91% ee) without subsequent racemization. PMID- 20024330 TI - Spontaneous and reversible self-assembly of a polypeptide fragment of insulin like growth factor binding protein-2 into fluorescent nanotubular structures. AB - In this communication, we report the spontaneous and reversible in vitro self assembly of a polypeptide fragment derived from the C-terminal domain of Insulin like Growth Factor Binding Protein (IGFBP-2) into soluble nanotubular structures several micrometres long via a mechanism involving inter-molecular disulfide bonds and exhibiting enhanced fluorescence. PMID- 20024331 TI - Inducing synthesis of amorphous EuFePt nanorods and their comprehensive enhancement of magnetism, thermostability and photocatalysis. AB - EuFePt ternary amorphous alloy nanorods are first synthesized through Eu itself inducing action, and this nanoalloy including 4f electrons exhibits excellent properties on magnetism, thermostability, especially the cooperation photocatalysis activity of TiO(2). PMID- 20024332 TI - Asymmetric synthesis of 2-substituted pyrrolidines by addition of Grignard reagents to gamma-chlorinated N-tert-butanesulfinyl imine. AB - A highly diastereoselective addition of various Grignard reagents to chiral gamma chlorinated N-tert-butanesulfinyl imine resulting in the formation of 2 substituted pyrrolidines is reported. This method is general and also efficient for the preparation of both enantiomers of 2-aryl, 2-alkyl and 2-vinyl substituted pyrrolidines in high yields. PMID- 20024333 TI - Supramolecular photocatalysis by confinement--photodimerization of coumarins within cucurbit[8]urils. AB - Cucurbit[8]uril (as low as 10 mol%) acts as a supramolecular catalytic nanoreaction vessel and facilitates the photodimerization of coumarins in water leading to syn dimers. Saturation kinetics shows a sigmoidal dependence with a turnover number of 3.4 min(-1) and a Hill constant of 1.8 indicating a co operative mechanism in the catalytic process. PMID- 20024335 TI - Single crystal structure of copper hexadecafluorophthalocyanine (F16CuPc) ribbon. AB - The single crystal structure of a micrometre-scale copper hexadecafluoro phthalocyanine (F(16)CuPc) ribbon synthesized by vaporization-condensation recrystallization (VCR) process was resolved by using a synchrotron X-ray diffractometer. PMID- 20024334 TI - Ionogels as drug delivery system: one-step sol-gel synthesis using imidazolium ibuprofenate ionic liquid. AB - Ionogels containing imidazolium ibuprofenate have been shown to be an efficient drug releasing system with kinetics controlled by the nature of the silica wall. PMID- 20024336 TI - Hydrophobic ionic liquids with strongly coordinating anions. AB - Ionic liquids containing the hexafluoroacetylacetonate anion are immiscible with water and they exhibit strong metal-complexing ability. PMID- 20024337 TI - Increasing the hydrophobicity and electrospray response of glycans through derivatization with novel cationic hydrazides. AB - Novel tags are used to increase the hydrophobicity of glycans and impart a permanent charge yielding as great as a approximately 5-fold increase in electrospray response from both a standard and complex mixture. PMID- 20024338 TI - Photonic interaction between quantum dots and gold nanoparticles in discrete nanostructures through DNA directed self-assembly. AB - Discrete nanostructures of CdSe@ZnS QDs and Au NPs were prepared and the photonic interactions between the QDs and Au NPs were systematically investigated. We found that the Au/QD ratio, separation distances between Au NPs and QDs, and the size of the Au NPs in a given discrete nanostructure all affect the interaction between Au NPs and QDs. PMID- 20024340 TI - Porous ionic/molecular crystal composed of highly symmetric magnetic clusters. AB - A porous ionic/molecular crystal composed of discrete triakis tetrahedral Co(8) clusters with an ideal T(d) symmetry shows interesting magnetism and porosity. PMID- 20024339 TI - Addressable electrochemiluminescence detection system based on redox-cycling of Ru(bpy)(3)(2+). AB - This is the first report on addressable electrochemiluminescence (ECL) based on redox-cycling of tris(2,2'-bipyridine)-ruthenium(II) (Ru(bpy)(3)(2+)). By changing the column or row electrodes addressed, the ECL at each address point can be detected separately. PMID- 20024341 TI - A liposome-based nanostructure for aptamer directed delivery. AB - A therapeutic aptamer conjugated liposome drug delivery system which delivered loaded drug to target cells with high specificity and excellent efficiency was prepared and characterized. PMID- 20024342 TI - Can nitrones functionalize carbon nanotubes? AB - An unprecedented functionalization of multi-walled carbon nanotubes (MWCNTs) has been conveniently achieved by the 1,3-dipolar cycloaddition of a cyclic nitrone. This organic functionalization yields materials with a great solubility in DMF (close to 10 mg per mL of DMF) preferentially occurring at the defects of the MWCNT sp(2) network. PMID- 20024343 TI - Synthesis of monodispersed nanoparticles functionalized carbon nanotubes in plasma-ionic liquid interfacial fields. AB - Metal nanoparticles intercalated into and encapsulated inside single-walled (SWNTs) and double-walled (DWNTs) carbon nanotubes are synthesized using a plasma technique combined with the introduction of ionic liquids under low gas pressures. Owing to the synthesis in nano-spaces of the SWNTs and DWNTs as a template, high-density and monodispersed metal nanoparticles are realized, which could be applied to specific composite-nanodevices based on carbon nanotubes. PMID- 20024344 TI - One-pot tandem 1,4-1,2-addition of phosphites to quinolines. AB - Trialkyl and silylated dialkyl phosphites were evaluated as phosphorus nucleophiles for the addition to quinolines in a strong acidic medium allowing consecutive 1,4- and 1,2-addition breaking up the aromatic stabilisation, thereby leading to 2,4-diphosphono-1,2,3,4-tetrahydroquinolines in one single reaction step in moderate to good yields (2-84%). PMID- 20024345 TI - Total synthesis of a library of designed hybrids of the microtubule-stabilising anticancer agents taxol, discodermolide and dictyostatin. AB - A hybrid library of the marine natural products dictyostatin and discodermolide, incorporating the taxol or taxotere side chains, were synthesised; preliminary biological evaluation in the PANC-1 cancer cell line revealed significant antiproliferative activity, demonstrating that a macrolide scaffold is an effective surrogate for the baccatin core of taxol. PMID- 20024346 TI - A click chemistry based coordination polymer inside small heat shock protein. AB - A branched iron-phenanthroline based coordination polymer has been constructed in a water based system using a click chemistry approach to link monomeric coordination complexes together within a protein cage nanoarchitecture, which acts both as a template and a sized constrained reaction environment. PMID- 20024347 TI - Stereoselective synthesis of trans-beta-lactams by palladium-catalysed carbonylation of vinyl aziridines. AB - The palladium-catalysed carbonylation of vinyl aziridines can give either the trans- or cis-beta-lactam preferentially or even the delta-lactam simply by adjusting the reaction parameters ([Pd], [CO], temperature). PMID- 20024348 TI - Palladium-catalyzed [3C + 2C + 2C] cycloaddition of enynylidenecyclopropanes: efficient construction of fused 5-7-5 tricyclic systems. AB - We report a Pd-catalyzed intramolecular [3C + 2C + 2C] cycloaddition between alkylidenecyclopropanes, alkynes and alkenes. The method provides synthetically relevant 5-7-5 tricyclic structures, with good chemoselectivity and complete diastereoselectivity. PMID- 20024349 TI - Dicationic and zwitterionic catalysts for the amine-initiated, immortal ring opening polymerisation of rac-lactide: facile synthesis of amine-terminated, highly heterotactic PLA. AB - Dicationic, zwitterionic and "conventional" yttrium compounds act as catalysts for the primary or secondary amine-initiated immortal ROP of rac-lactide; amine terminated, highly heterotactic poly(rac-lactides) with narrow polydispersities and well-controlled molecular weights are prepared in this manner. PMID- 20024350 TI - Efficient protein renaturation using tunable hemifluorinated anionic surfactants as additives. AB - We show that hemifluorinated anionic surfactants are effective as renaturing additives for bovine carbonic anhydrase (CAB) below their critical micelle concentration (CMC) providing high refolding yields for thermally and chemically denatured protein. PMID- 20024351 TI - A seven atom cluster in a carbon cage, the crystallographically determined structure of Sc4(mu3-O)3@Ih-C80. AB - The tetrahedral array of four scandium atoms with oxygen atoms capping three of the four faces found in Sc(4)(mu(3)-O)(3)@I(h)-C(80) is the largest cluster isolated to date inside a fullerene cage. PMID- 20024352 TI - Synthesis of CuO on mesoporous silica and its applications for coupling reactions of thiols with aryl iodides. AB - Novel CuO on mesoporous silica is prepared under a convenient approach, and has been shown to be an efficient catalyst for cross-coupling reactions of thiols with aryl iodides with only 1.0-5.0 mol% catalyst loading. PMID- 20024353 TI - Comparative spectroscopic studies on porphyrin derivatives: electronic perturbation of N-confused and N-fused porphyrins. AB - N-confused and N-fused porphyrins revealed quite unique photophysical properties arising from their perturbed pi-conjugation pathways induced by structural modifications such as confusion and fusion of pyrrole rings compared with their parent porphyrin molecules. PMID- 20024354 TI - Molecular recognition of halogen-tagged aromatic VOCs at the air-silicon interface. AB - Selective and reversible complexation of halogen-tagged aromatic VOCs by a quinoxaline cavitand-decorated Si surface is demonstrated. The specific host guest interactions of the Si-bonded receptors are proved to be responsible of the surface recognition properties, while extracavity non specific adsorptions are totally suppressed compared to the bulk material. PMID- 20024355 TI - Targeted synthesis of a 3D porous aromatic framework for selective sorption of benzene. AB - A novel 3D porous aromatic framework (PAF) based on a tetraphenylmethane block and a triangular triazine ring been designed and synthesized, with 1109 m(2) g( 1) Langmuir surface area, and shows selective sorption of benzene. PMID- 20024357 TI - Heteromeric double helix formation by cross-hybridization of chloro-and fluoro substituted quinoline oligoamides. AB - Oligoamides of 8-chloroquinoline have been synthesized and shown to assemble into double helical dimers both in solution and in the solid state, and to undergo cross-hybridization with 8-fluoroquinoline oligoamide analogues; the handedness of these double helices can be controlled via chiral residues. PMID- 20024356 TI - Biocompatibility of choline salts as crosslinking agents for collagen based biomaterials. AB - A series of novel choline based salts, some of which can be described as ionic liquids, are prepared and evaluated for their biocompatibility; when combined with collagenous biomaterials they exhibit good cell viability and adhesion properties as required for biomedical implant applications. PMID- 20024358 TI - Flexible synthesis, structural determination, and synthetic application of a new C1-symmetric chiral ammonium betaine. AB - A C(1)-symmetric chiral ammonium betaine has been developed, and its intramolecular ion-pairing structure in solid state and its catalytic performance to achieve the highly stereoselective Mannich-type reaction of 2-alkoxythiazol 5(4H)-ones are revealed. PMID- 20024359 TI - Synthesis of nanostructured silicon carbide spheres from mesoporous C-SiO2 nanocomposites. AB - Nanostructured porous silicon carbide hollow spheres have been synthesized by phase separation in mesoporous silica-carbon nanocomposites at high temperatures and subsequent carbothermal reduction and crystallization. PMID- 20024360 TI - Enormous enhancement in photocurrent generation using electrochemically fabricated gold nanostructures. AB - A self-assembled monolayer of porphyrins fabricated on the surface of an electrochemically deposited gold nanostructure exhibits enormous enhancement of photocurrent due to porphyrin excitation, especially in the near-infrared region, where localized surface plasmon resonance was responsible. PMID- 20024361 TI - Novel AAO films and hollow nanostructures fabricated by ultra-high voltage hard anodization. AB - Novel anodic aluminium oxide (AAO) films and hollow nanostructures were synthesized via a simple electrochemical and chemical etching route; fluctuating nanotube growth inside AAO film fabricated under ultra-high voltage was considered to be the main reason for the formation of such new structures. PMID- 20024362 TI - PPh3-catalyzed [2 + 2 + 2] and [4 + 2] annulations: synthesis of highly substituted 1,2-dihydropyridines (DHPs). AB - PPh(3)-catalyzed [2 + 2 + 2] annulations between two units of an activated terminal alkyne and one unit of an aryl N-tosylimine have been developed to provide a synthetic method for highly substituted dihydropyridines. On the basis of the mechanism of the [2 + 2 + 2] annulation, PPh(3)-catalyzed [4 + 2] annulations have also been discovered. PMID- 20024363 TI - Rhodium-mediated activation of an alkane-type C-H bond. AB - Abnormal C4-bonding of N-heterocyclic carbenes effectively modulates the electron density at rhodium and allows for the selective cleavage of an unactivated C(sp(3))-H bond, whereas no such intramolecular C-H bond breaking is observed when the carbene binds normally through the C2 carbon. PMID- 20024364 TI - A facile synthesis of dibenzopyrroloazepinones as tetracyclic allocolchicinoids- an unusual 1,2-phenyl shift. AB - A facile synthesis of dibenzopyrroloazepinones via an electrophilic cyclisation of a biphenyl-acyliminium ion is described; an unusual 1,2-phenyl shift occurs when the C-1' carbon is the more nucleophilic than the C-2' carbon. PMID- 20024365 TI - Chiral bifunctional phase transfer catalysts for asymmetric fluorination of beta keto esters. AB - Chiral bifunctional phase transfer catalysts introducing bis(diarylhydroxymethyl) substituents at 3,3'-positions of the chiral binaphthyl core were successfully applied to asymmetric fluorination of cyclic beta-keto esters with high enantioselectivities. PMID- 20024366 TI - Template-controlled synthesis of a planar [16]ane-P2C(NHC)2 macrocycle. AB - Reaction of tetraphosphine platinum(II) complexes with 2-azidoethyl isocyanide 3 in methanol leads to complexes with two trans-coordinated NH,NH-stabilized carbene ligands 6(X)(2). Complexes 6(X)(2) react with phenyldivinylphosphine under substitution of the remaining phosphine ligands followed by an intramolecular hydroamination reaction to produce complex 1(PF(6))(2) with a macrocyclic [16]ane-P(2)C(NHC)(2) ligand. PMID- 20024367 TI - Organocatalytic asymmetric Povarov reactions with 2- and 3-vinylindoles. AB - The asymmetric Povarov reaction of N-arylimines with 2- and 3-vinylindoles has been developed using a chiral phosphoric acid ((S)-TRIP) as catalyst. The peculiar reactivity of vinylindoles allowed also the disclosure of a Povarov Friedel-Crafts sequence, and the trapping of the reaction intermediate with nucleophilic species, thus providing a versatile platform for the preparation of highly enantioenriched indole derivatives. PMID- 20024368 TI - A kit for the investigation of live Escherichia coli cell adhesion to glycosylated surfaces. AB - A combination of microtiter plate functionalization techniques and two facile bacterial adhesion inhibition assays form a flexible toolbox for the investigation of bacterial adhesion mechanisms on glycosylated surfaces. PMID- 20024369 TI - 13C labeling reveals multiple amination reactions in the biosynthesis of a novel polyketide polyamine antibiotic zeamine from Dickeya zeae. AB - The structure of zeamine, a novel polyamino-amide antibiotic metabolite of Dickeya zeae has been established by NMR and detailed MS analyses; labelling studies with (13)C-labelled acetates suggest that the repeating secondary amine containing motif may be biosynthesised via a modular PKS containing aminotransferase domains. PMID- 20024370 TI - Catalytic SNAr reaction of non-activated fluoroarenes with amines via Ru eta6 arene complexes. AB - Ru-catalyzed S(N)Ar reaction of non-activated fluoroarenes with secondary amines proceeded through eta(6)-arene complexes to give aminated products in up to 79% yield. PMID- 20024371 TI - A zwitterionic zirconium complex that catalyzes hydroamination of aminoalkenes at room temperature. AB - The zwitterionic cyclopentadienyl-bis(2-oxazolinyl)borate diamidozirconium(IV) complex is a precatalyst for the cyclization of aminoalkenes to five- and six membered rings under mild conditions. PMID- 20024374 TI - Lanthanide ions as spectral converters for solar cells. AB - The use of lanthanide ions to convert photons to different, more useful, wavelengths is well-known from a wide range of applications (e.g. fluorescent tubes, lasers, white light LEDs). Recently, a new potential application has emerged: the use of lanthanide ions for spectral conversion in solar cells. The main energy loss in the conversion of solar energy to electricity is related to the so-called spectral mismatch: low energy photons are not absorbed by a solar cell while high energy photons are not used efficiently. To reduce the spectral mismatch losses both upconversion and downconversion are viable options. In the case of upconversion two low energy infrared photons that cannot be absorbed by the solar cell, are added up to give one high energy photon that can be absorbed. In the case of downconversion one high energy photon is split into two lower energy photons that can both be absorbed by the solar cell. The rich and unique energy level structure arising from the 4f(n) inner shell configuration of the trivalent lanthanide ions gives a variety of options for efficient up- and downconversion. In this perspective an overview will be given of recent work on photon management for solar cells. Three topics can be distinguished: (1) modelling of the potential impact of spectral conversion on the efficiency of solar cells; (2) research on up- and downconversion materials based on lanthanides; and (3) proof-of-principle experiments. Finally, an outlook will be given, including issues that need to be resolved before wide scale application of up- and downconversion materials can be anticipated. PMID- 20024375 TI - What are the factors that control non-aqueous/AOT/n-heptane reverse micelle sizes? A dynamic light scattering study. AB - Polar solvent-AOT interactions control non-aqueous/AOT/n-heptane reverse micelle polar core sizes. PMID- 20024376 TI - Microporous carbon coated silicon core/shell nanocomposite via in situ polymerization for advanced Li-ion battery anode material. AB - A microporous carbon coated core/shell Si@C nanocomposite prepared by in situ polymerization exhibits a stable capacity of over 1200 mAh g(-1) with 95.6% retention even after 40 cycles, which makes it a promising anode material for lithium ion batteries. PMID- 20024377 TI - The role of interstitial H2 in hydrogen diffusion in light metal borohydrides. AB - First principles calculations suggest that mobility of H in solid borohydrides is dominated by neutral interstitial H(2), not charged defects. PMID- 20024378 TI - Fischer-Tropsch synthesis of liquid fuels: learning lessons from homogeneous catalysis. AB - Herein, we present results opposing the so-called carbide mechanism in the Fischer-Tropsch synthesis of liquid fuels and point out analogies between this heterogeneously catalysed process and related homogeneous processes. PMID- 20024379 TI - Prediction and characterization of the single-electron sodium bond complexes Y C...Na-H [Y = H3, H3CH2, (H3C)2H and (H3C)3]. AB - The prediction and characterization of the single-electron sodium bond complexes Y-C...Na-H [Y = H(3), H(3)CH(2), (H(3)C)(2)H and (H(3)C)(3)] have been investigated for the first time by using MP2/6-311++G(d,p), MP2/6-311++G(2d,2p) and MP2/aug-cc-pVDZ methods. The strength of the interactions in H(3)C...Na-H, H(3)CH(2)C...Na-H, (H(3)C)(2)HC...Na-H, and (H(3)C)(3)C...Na-H complexes has been analyzed. It is shown that the (H(3)C)(3)C radical with Na-H forms the strongest single-electron sodium bond, followed by the (H(3)C)(2)HC radical and then the H(3)CH(2)C radical. H(3)C radical forms the weakest single-electron sodium bond. NBO and AIM analyses have also been used to estimate such conclusions. Furthermore, there are few linear/nonlinear relationships among the several parameters in system and the interaction mode of single-electron Na bond is LP(1)(C) --> LP(1)*(Na), which is different from the single-electron H bond and single electron halogen bond. By comparisons with some related systems, it is concluded that the strength of single-electron bond is increased in the order: hydrogen bond < sodium bond < bromine bond < lithium bond. PMID- 20024380 TI - Theoretical analysis of carbon nanotube wetting in polystyrene nanocomposites. AB - Besides chemical functionalisation, the use of surfactants can be applied to debundle and disperse carbon nanotubes before further application in polymer nanocomposites. In this work we present a theoretical analysis of the interaction between single-walled carbon nanotubes and sodium dodecyl sulfate as surfactant and/or polystyrene as polymer matrix using semi-empirical AM1 calculations. Results indicate that the use of short potassium sulfate-terminated polystyrene chains as an extra component can help to remove the surfactant from the nanotube surface within the matrix, resulting in improved electronic properties of the nanocomposite. PMID- 20024381 TI - Molecular dynamics simulation of nanoconfined glycerol. AB - We present results from molecular dynamics simulations of liquid glycerol confined in a realistic model of a cylindrical silica nanopore. The influence of the hydrophilic surface and the geometrical confinement on the structure, hydrogen-bond lifetime, rotational and translational molecular dynamics are analysed. Layering and dynamical heterogeneities are induced by confinement. These features share some similarities with previous observations in simpler van der Waals glass-forming liquids. In addition, the specificity of glycerol as an associated liquid shows up in confinement by the formation of interfacial hydrogen bonds and some modifications of the in-pore hydrogen-bonding network. Confinement is also seen to influence the relaxation dynamics and the glassy behaviour in the supercooled state. These phenomena revealed by molecular simulation are important inputs for a better understanding of the many recent experimental results on confined glycerol and more generally for the possible manipulation of associated liquids in porous or fluidic devices. PMID- 20024382 TI - Redox-induced partner radical formation and its dynamic balance with radical dimer in cucurbit[8]uril. AB - N-(4-Hydroxy-phenoxyethyl)-N'-ethyl-4,4'-bipyridium (1) can form a stable 1 : 1 inclusion complex with CB[8] in aqueous solution, in which the hydroxyphenol (HP) moiety is back-folded and inserted together with the viologen moiety into the cavity of CB[8]. When the ethyl viologen dication (EV(2+)) in 1 is reduced, chemically or electrochemically, an intramolecular partner radical (EV(+)* HP)/CB[8] can be detected, meanwhile, a dynamic balance between the partner radical and the intermolecular radical dimer (EV(+)*-HP)(2)/CB[8] can be observed. PMID- 20024383 TI - In situ STM investigation of the lithium underpotential deposition on Au(111) in the air- and water-stable ionic liquid 1-butyl-1-methylpyrrolidinium bis(trifluoromethylsulfonyl)amide. AB - In the present paper the underpotential deposition (UPD) of lithium on Au(111) from 0.5 mol L(-1) LiTFSA in the air- and water stable ionic liquid 1-butyl-1 methylpyrrolidinium bis(trifluoromethylsulfonyl)amide, [Py(1,4)]TFSA, has been investigated by cyclic voltammetry (CV) and in situ scanning tunneling microscopy (STM). The pure [Py(1,4)]TFSA was found to be inert in the potential regime investigated. The results show that the lithium UPD on Au(111) in [Py(1,4)]TFSA begins at potentials considerably positive to the electrode potential of bulk deposition and follows a layer-by-layer mechanism with the formation of at least two monolayers. A large number of monoatomically deep pits appear when the potential reaches positive values, which is an indication that a Li-Au alloy was formed. PMID- 20024384 TI - Para-hydrogen induced polarization in multi-spin systems studied at variable magnetic field. AB - A theoretical description of para-hydrogen-induced polarization (PHIP) is developed, applicable to coupled multi-spin systems that are polarized at an arbitrary magnetic field. Scalar spin-spin interaction is considered to be the leading factor governing PHIP formation and transfer. At low magnetic fields, these interactions make the spins strongly coupled and cause efficient, coherent re-distribution of spin polarization. We describe the effects of strong coupling and field cycling for a three-spin system and compare calculated spectra with the experimental examples available. By using a fast field-cycling device, which shuttles the whole NMR probe, and thereby makes high-resolution NMR detection at high field possible, we studied PHIP patterns for a set of different fields between 0.1 mT and 7 T. PHIP spectra were measured for ethylbenzene as the product of a catalytic reaction between para-hydrogen and styrene. Additionally, the polarizations of ethylbenzene bound to the catalyst, and of the starting styrene molecule were analyzed. This is the first time that the full field dependence of PHIP has been determined experimentally. The spectra obtained are in perfect agreement with the simulations for the CH(2) and CH(3) protons of ethylbenzene and even for its weakly-polarized aromatic protons. Analysis of styrene polarization shows that the time profile of the field variation has pronounced effects on the PHIP pattern. Our study gives evidence that scalar spin spin interactions determine the PHIP patterns. Possible applications of the theory are discussed. PMID- 20024385 TI - On the low volatility of cyclic esters: an infrared spectroscopy comparison between dimers of gamma-butyrolactone and methyl propionate. AB - The dramatically lower volatility of gamma-butyrolactone compared to its open chain analog methyl propionate is analyzed at the molecular dimer level using FTIR spectroscopy in supersonic jets. It is found that the spectral shifts from the monomer to the dimer are about three times more pronounced in the lactone at low temperatures. The spectra are consistent with sandwich-like dimers optimizing their strong dipole-dipole interaction, possibly augmented by specific C-H...O=C hydrogen bond contacts. The spectra show significant evolution from the dimer to the condensed phase, indicative of secondary interactions with the ester oxygen and long range forces. The reduced dipole moment in the open chain ester leads to less specific interactions, unless a trans conformation of the ester group as in the lactones is enforced. The latter is not energetically accessible in open chain esters because it would bring the molecular C=O and C-O-C dipole moments into an unfavorable near-parallel orientation, thus their higher volatility. PMID- 20024386 TI - Temperature evolution of structure and bonding of formic acid and formate on fully oxidized and highly reduced CeO2(111). AB - Adsorption of formate on oxide surfaces plays a role in water-gas shift (WGS) and other reactions related to H(2) production and CO(2) utilization. CeO(2) is of particular interest because its reducibility affects the redox of organic molecules. In this work, the adsorption and thermal evolution of formic acid and formate on highly ordered films of fully oxidized CeO(2)(111) and highly reduced CeO(x)(111) surfaces have been studied using reflection absorption infrared spectroscopy (RAIRS) under ultra-high vacuum conditions, and the experimental results are combined with density functional theory (DFT) calculations to probe the identity, symmetry, and bonding of the surface intermediates. Disordered ice, ordered alpha-polymorph and molecular formic acid bonded through the carbonyl are observed at low temperatures. By 250 K, desorption and deprotonation lead to formate coexisting with hydroxyl on CeO(2)(111), identified to be a bridging bidentate formate species that is coordinated to Ce cations in nearly C(2v) symmetry and interacting strongly with neighboring H. Changes in the spectra at higher temperatures are consistent with additional tilting of the formate, resulting in C(s)(2) or lower symmetry. This change in bonding is caused primarily by interaction with oxygen vacancies introduced by water desorption at 300 K. On reduced CeO(x), multiple low-symmetry formate states exist likewise due to interactions with oxygen vacancies. Isotopic studies demonstrate that the formyl hydrogen does not contribute to H incorporated in hydroxyl on the surface, and that both formate oxygen atoms may exchange with lattice oxygen at 400 K. The combined experimental and theoretical results thus provide important insights on the surface reaction pathways of formic acid on ceria. PMID- 20024387 TI - Influence of flocculation and coalescence on the evolution of the average radius of an O/W emulsion. Is a linear slope of R3 vs. t an unmistakable signature of Ostwald ripening? AB - The LSW theory of Ostwald ripening, predicts a linear variation of the cube of the average radius of a dispersion as a function of time (R(3)vs. t) [I. M. Lifshitz, V. V. Slyozov, J. Phys. Chem. Solids, 1961, 19, 35-50; C. Wagner, Z. Elektrochem., 1961, 65, 581-591]. It also envisages a left-skewed drop-size distribution with a cut-off radius of 1.5R. Consequently, non-linear changes of R(3)vs. t are usually ascribed to either a transient period of time (previous to the attainment of the asymptotic limit of ripening) or other destabilisation processes. Up to now the effect of Brownian motion on Ostwald ripening (OR) has not been considered, although it is by far the strongest limitation of the LSW theory. In this work we show the results of incorporating the algorithm of De Smet et al. for Ostwald ripening simulations [Y. De Smet, L. Deriemaeker, R. Finsy, Langmuir, 1997, 13, 6884-6888] to our emulsion stability simulations (ESS) code. In particular, the short-time evolution of a dilute dodecane/water nanoemulsion in the absence of stabilisers is studied. At high ionic strength, the simulations suggest that R(3) can change linearly with time during the transient period of Ostwald ripening, due to the flocculation and the coalescence of the drops. This behavior is confirmed by the experiments for t < 100 s. At low ionic strength a concave downward curve is observed, both theoretically and experimentally. PMID- 20024388 TI - The method of local increments for the calculation of adsorption energies of atoms and small molecules on solid surfaces. Part I. A single Cu atom on the polar surfaces of ZnO. AB - The method of local increments is used in connection with the supermolecule approach and an embedded cluster model to calculate the adsorption energy of single Cu atoms at different adsorption sites at the polar surfaces of ZnO. Hartree-Fock calculations for the full system, adsorbed atom and solid surface, and for the fragments are the first step in this approach. In the present study, restricted open-shell Hartree-Fock (ROHF) calculations are performed since the Cu atom possesses a singly-occupied 4s orbital. The occupied Hartree-Fock orbitals are then localized by means of the Foster-Boys localization procedure. The correlation energies are expanded into a series of many-body increments which are evaluated separately and independently. In this way, the very time-consuming treatment of large systems is replaced with a series of much faster calculations for small subunits. In the present application, these subunits consist of the orbitals localized at the different atoms. Three adsorption situations with rather different bonding characteristics have been studied: a Cu atom atop a threefold-coordinated O atom of an embedded Zn(4)O(4) cluster, a Cu atom in an O vacancy site at the O-terminated ZnO(000-1) surface, and a Cu atom in a Zn vacancy site at the Zn-terminated ZnO(0001) surface. The following properties are analyzed in detail: convergence of the many-body expansion, contributions of the different n-body increments to the adsorption energy, treatment of the singly occupied orbital as "localized" or "delocalized". Big savings in computer time can be achieved by this approach, particularly if only the localized orbitals in the individual increment under consideration are described by a large correlation adapted basis set, while all other orbitals are treated by a medium-size Hartree Fock-type basis set. In this way, the method of local increments is a powerful alternative to the widely used methods like DFT or RI-MP2. PMID- 20024389 TI - A pi-stacked phenylacetylene and 1,3,5-triazine heterodimer: a combined spectroscopic and ab initio investigation. AB - The IR-UV double resonance spectroscopy of a complex between phenylacetylene and 1,3,5-triazine reveals that the acetylene C-H group of phenylacetylene is minimally perturbed due to its interaction with 1,3,5-triazine. Further, the IR spectrum clearly indicates that 1,3,5-triazine primarily interacts with pi electron density of the benzene ring in phenylacetylene. Geometries obtained at the DFT/MO6-2X and MP2/aug-cc-pVDZ levels, combined with highly accurate energy calculations at the complete basis set (CBS) limit of CCSD(T), establish formation of the displaced pi-stacked heterodimer between phenylacetylene and 1,3,5-triazine. PMID- 20024390 TI - Equation of state for hard-sphere fluids offering accurate virial coefficients. AB - The asymptotic expansion method is extended by using currently available accurate values for the first ten virial coefficients for hard sphere fluids. It is then used to yield an equation of state for hard sphere fluids, which accurately represents the currently accepted values for the first sixteen virial coefficients and compressibility factor data in both the stable and the metastable regions of the phase diagram. PMID- 20024391 TI - Valence shell electronic spectroscopy of isoprene studied by theoretical calculations and by electron scattering, photoelectron, and absolute photoabsorption measurements. AB - The first ab initio calculations (vertical energies and oscillator strengths) are reported for the neutral electronic transitions of isoprene (2-methyl-1,3 butadiene), CH(2)CHC(CH(3))CH(2). The VUV photoabsorption spectroscopy of the molecule is presented in the energy range 4.6 to 10.8 eV (270-125 nm) with the highest resolution yet reported above 6.05 eV, revealing new spectral features. Valence and Rydberg transitions have been assigned in accordance with the theoretical results and the associated vibronic series have been analysed. The absolute photoabsorption cross sections at energies below 6.89 eV have been used to calculate the photolysis lifetime of isoprene in the upper stratosphere (20-50 km). Electron energy loss spectroscopy (EELS) measurements have enabled further photoabsorption cross sections to be derived in the range 9-28 eV. The first ab initio calculations have been carried out to determine excitation energies to the lowest energy ionic states of isoprene. The calculations are compared with the He(i) photoelectron spectrum (8 to 17 eV) and new vibrational structure is observed in the first photoelectron band. PMID- 20024392 TI - A computational study of the nuclear magnetic resonance parameters for double proton exchange pathways in the formamide-formic acid and formamide-formamidine complexes. AB - In this paper, we present density functional theory calculations to predict the NMR parameters for two model systems: the formamide-formic acid (FM...FA) and formamide-formamidine (FM...FI) complexes, where intermolecular double proton exchange occurs. For the first time, the NMR parameters have been calculated along the reaction paths of the proton transfers described by means of the intrinsic reaction coordinate (IRC) procedure. The most interesting one-bond spin spin coupling constants, (1(h))J(XH), between migrating protons and heavier nuclei change character from intra- to intermolecular along the pathway. The maximal positive values of the reduced (1(h))K(XH) coupling constants correspond to the situation when they are intramolecular; they decrease along the path, change sign and reach small negative values, becoming intermolecular couplings. The differing character of the double proton exchange resulting from the synchronicity or asynchronicity of the process is reflected in the calculated NMR parameters. Surprisingly substantial values have been calculated for the six-bond intermolecular proton-proton (6h)J(HH) coupling constants between protons bound to the carbon atoms. A simple procedure consisting of removal of the proton(s) forming the hydrogen bonds has been employed to indicate an influence of hydrogen bonding on the intermolecular coupling constants. Some of the spin-spin coupling constants ((2h)J(XY)) are predominantly transmitted through hydrogen bonds and decrease with removal of the proton(s), while others ((4h)J(CC)) are less sensitive to the presence or absence of the protons of hydrogen bonding. PMID- 20024393 TI - Impedance spectra of mixed conductors: a 2D study of ceria. AB - In this paper we develop an analytical framework for the study of electrochemical impedance of mixed ionic and electronic conductors (MIEC). The framework is based on non-equilibrium thermodynamics and it features the coupling of electrochemical reactions, surface transport and bulk transport processes. We utilize this work to analyze two-dimensional systems relevant for fuel cell science via the finite element method (FEM). Alternate current impedance spectroscopy (AC-IS or IS) of a ceria symmetric cell is simulated near equilibrium conditions (zero bias) for a wide array of working conditions including variations of temperature and H(2) partial pressure on a two-dimensional doped ceria sample with patterned metal electrodes. The model shows agreement between computed IS curves and the experimental literature where the relative error on the impedance is consistently below 2%. Important two-dimensional effects such as the impact of thickness decrease and the influence of variable electronic and ionic diffusivities on the impedance spectra are also explored. PMID- 20024394 TI - Silver nanoparticles behave as hydrophobic solutes towards the liquid water structure in the interaction shell. A Raman study in the O-H stretching region. AB - Weak interactions, never reported yet, between silver nanocolloids and water, have been studied by Raman spectroscopy in the O-H stretching region. Subtle interactions regarding the first hydration shell, and perturbing only at a short range the bulk liquid structure of the solvent have been revealed. The interface interactions cause the O-H stretching band of water in the interface layer spectrum to be shifted at higher wavenumbers with respect to the neat water Raman signals. Order/disorder ratio variations, with respect to the liquid spectrum of pure water, are reported. PMID- 20024395 TI - A quantum mechanical study of TiCl3 alpha, beta and gamma crystal phases: geometry, electronic structure and magnetism. AB - The electronic structure of different magnetic states of alpha, beta and gamma modifications of TiCl(3) has been computed employing the density functional theory with periodic boundary conditions and localized Gaussian basis sets. The analysis of the density of the electronic states (DOS) and of the spin density makes it possible to classify these halides as Mott-Hubbard insulators, where the band gap appears a result of large on-site Coulomb interaction. For each crystalline phase, the relative stability of different magnetic states has been analyzed in terms of exchange mechanisms. The electronic population data along with the spin density maps support the assumption of a d(1) Titanium ion in a distorted octahedral crystal field, notwithstanding the not fully ionic character of TiCl(3) modifications. Dispersion forces are particularly important for this material: a classical correction (of the type f(R)/R(6)) has been added to the DFT energies and gradients, providing a good agreement with structural data. PMID- 20024396 TI - A TD-DFT investigation of ground and excited state properties in indoline dyes used for dye-sensitized solar cells. AB - The ground and excited state properties of three indoline dyes, namely D102, D131 and D149, especially designed for dye sensitized solar cell (DSC) applications have been studied by the means of density functional theory (DFT) and time dependent DFT (TD-DFT) and compared with experimental absorption and fluorescence spectra. By comparison, insight on the behavior of a related dye (D205) is also given. Beside the good agreement between the computed and experimental spectra further proving the accuracy of the method used, that is, a hybrid (here PBE0) exchange correlation functional in conjunction with a polarizable continuum solvent (PCM) model and a medium size basis set, the calculations allow new insights into the electronic structure of this family of indolines, and in particular the electronic and geometrical structure at the first excited state. Possible insights on the optimization of dyes for photovoltaics applications, as well as warnings on the extrapolation of isolated dye properties towards cells behavior are given. PMID- 20024397 TI - Adsorption-desorption and photocatalytic properties of inorganic-organic hybrid cadmium thiosulfate compounds. AB - Three inorganic-organic hybrid framework cadmium thiosulfate phases have been investigated for adsorption and photodegradation of organic dye molecules. Different classes of organic dyes, viz., triaryl methane, azo, xanthene, anthraquinone, have been studied. The anionic dyes with sulfonate groups appear to readily adsorb on the cadmium thiosulfate compounds in an aqueous medium. The adsorption of the dye molecules, however, does not create any structural changes on the cadmium thiosulfate compounds, though weak electronic interactions have been observed. The adsorbed dyes have been desorbed partially in an alcoholic medium, suggesting possible applications in scavenging specific anionic dyes from the aqueous solutions. Langmuir adsorption/desorption isotherms have been used to model this behavior. UV-assisted (lambda(max) = 365 nm) photocatalytic decomposition studies on the cationic dyes indicate reasonable activity comparable with that of Degussa P-25 (TiO(2)) catalyst. Sunlight assisted photocatalytic studies have been carried out in detail employing hybrid framework compounds. The Langmuir-Hinshelwood kinetics model, employed to follow the degradation profile of the organic dyes, indicates that the photocatalytic degradation follows the order: triaryl methane > azo > xanthene. PMID- 20024398 TI - LOBA: a localized orbital bonding analysis to calculate oxidation states, with application to a model water oxidation catalyst. AB - We propose a method for calculation of oxidation states in transition metal complexes, utilizing a bonding analysis based on localized molecular orbitals in conjunction with traditional population analyses. The localized orbital bonding analysis (LOBA) is seen to accurately produce both the oxidation state and chemically intuitive views of bonding in the complexes studied. This is in contrast to simple population analyses where the oxidation states are not reproduced for even simple systems and more complex analyses which break down on problematic systems. We report the application to a manganese complex with potential activity as oxygen-evolving catalyst, determining the location of the oxidations. PMID- 20024399 TI - Directed assembly of Au and Fe nanoparticles on a TiOx/Pt(111) ultrathin template: the role of oxygen affinity. AB - The essential role of O affinity in the directed assembly of size-selected Au and Fe nanoparticles (NPs) on a TiO(x)/Pt(111) ultrathin oxide phase, an effective template for size selected metal NP growth, is revealed through scanning tunneling microscopy and density-functional calculations. A weakly interacting element (Au) diffuses rapidly and gets trapped in the vacancy defects (picoholes) located inside parallel rows (troughs, spaced 1.44 nm apart) peculiar to the film structure, producing size-selected NPs arranged in regular linear arrays aligned along the troughs. In contrast, an element with greater O affinity (Fe) experiences higher diffusion barriers, and the growth is dominated by kinetic effects, with a less effective preferential nucleation and the appearance of irregular NP morphologies. PMID- 20024400 TI - Growth of boehmite particles in the presence of xylitol: morphology oriented by the nest effect of hydrogen bonding. AB - The ability to design nanoparticles size and shape through the addition of simple and commercially available organic molecules is of particular interest in the catalytic domain because huge amounts of very fine powders are needed. The origin of this effect is all the more difficult to elucidate because the involved interactions are weak. In this paper, we have investigated the shaping of boehmite AlO(OH) nanoparticles in the presence of polyols like xylitol (C(5) alditol) by a combined experimental and theoretical approach. Experimental techniques such as XRD, TEM, IEP measurements, adsorption isotherms measurements, and (13)C MAS NMR experiments demonstrate that the effect of xylitol has a thermodynamic origin and suggest weak interactions between xylitol and the surface. Furthermore, the strongest proportion of lateral faces ((100), (001), and (101)) that of basal face would be in agreement with a preferential adsorption upon lateral surfaces. These results were refined by a computational approach. DFT calculations of surface energies (taking into account temperature and solvation effects) and of NMR shielding constants corroborate that molecular adsorption mode is preferred over all adsorption modes involving exchanges with surface OH groups. The preferred adsorption on lateral surfaces is attributed to the nest effect induced by hydroxyl groups localized on the concavities of the (001) and (101) surfaces, able to stabilize the xylitol molecule by hydrogen bonding, whereas the basal (010) surface is almost flat. This combined experimental and computational approach thus provides interesting rationalization for the morphology effects observed. PMID- 20024401 TI - Molecular dynamics simulations of polystyrene brushes in dry conditions and in toluene solution. AB - The properties of polystyrene brushes in dry conditions and in toluene solution are studied as a function of grafting density using molecular dynamics simulations. Both, individual brushes and double layers of opposing brushes are considered, the structural properties of which were found to be similar. The density profiles show very pronounced density oscillations which extend up to approximately 1.8 nm and fall into two groups of three peaks each. These features are observed regardless of grafting density and solvent conditions. In the absence of solvent, the chains undergo a transition from an oblate to a spherical shape as the grafting density increases. In contrast, in good solvent, the chains remain spherical independent of the grafting density. Solvation also increases the extension of the polystyrene chains roughly by a factor 2.5. Isotropic and two-dimensional radial distribution functions are used to characterize the structure of the polystyrene brushes. Toluene is observed to form up to four layers at the base of the grafted chains irrespective of grafting density. PMID- 20024402 TI - Manipulation and characterisation of accumulation and coarse mode aerosol particles using a Bessel beam trap. AB - Micron and sub-micron sized aerosol particles are captured, manipulated and characterised in a Bessel beam optical trap. Bright field microscopy and elastic light scattering measurements are used in combination to interrogate trapped particles and explore the optical landscape of the trap. We conclude that the Bessel trap has a number of advantages over optical tweezers in terms of characterisation of accumulation mode particles, manipulation of particles over macroscopic length scales and effective control of the gas phase. As such, the Bessel trap is a valuable addition to the aerosol optical toolkit. PMID- 20024403 TI - Pathways for hydrogen bond switching in a tetrameric methanol cluster. AB - Computational techniques (second order Moller-Plesset MP2 perturbation theory in conjunction with medium and large size basis sets) are applied to explore structural aspects of a hydrogen-bonded tetrameric cluster of methanol molecules, based geometrically on a tetrahedral arrangement of the four oxygen atoms of the cluster. The hydrogen-bonded structures that represent minima on the potential energy surface are established, and the complete set of pathways that allow interconversion between these structures through "switching" of the hydrogen bonding arrangement are elucidated. The implications of these results in terms of dynamic properties of the cluster are discussed. PMID- 20024405 TI - Static, rheological and mechanical properties of polymer nanocomposites studied by computer modeling and simulation. AB - Polymer nanocomposites (PNCs) often exhibit excellent mechanical, thermal, electrical and optical properties, because they combine the performances of both polymers and inorganic or organic nanoparticles. Recently, computer modeling and simulation are playing an important role in exploring the reinforcement mechanism of the PNCs and even the design of functional PNCs. This report provides an overview of the progress made in past decades in the investigation of the static, rheological and mechanical properties of polymer nanocomposites studied by computer modeling and simulation. Emphases are placed on exploring the mechanisms at the molecular level for the dispersion of nanoparticles in nanocomposites, the effects of nanoparticles on chain conformation and glass transition temperature (T(g)), as well as viscoelastic and mechanical properties. Finally, some future challenges and opportunities in computer modeling and simulation of PNCs are addressed. PMID- 20024406 TI - Gas phase folding of an (Ala)4 neutral peptide chain: spectroscopic evidence for the formation of a beta-hairpin H-bonding pattern. AB - IR and UV laser spectroscopy of an Ala-based 4-residue model peptide recorded under gas phase isolated conditions provides evidence for the intrinsic stability of compact folded structures resembling the extremity of a beta-hairpin, with a C(14) H-bond bridging the two ends of the chain, and enables us to assess the capabilities of new quantum chemistry techniques to account for dispersive interactions in a medium-size molecule. PMID- 20024407 TI - Computational identification of a metal organic framework for high selectivity membrane-based CO2/CH4 separations: Cu(hfipbb)(H2hfipbb)0.5. AB - The identification of membrane materials with high selectivity for CO(2)/CH(4) mixtures could revolutionize this industrially important separation. We predict using computational methods that a metal organic framework (MOF), Cu(hfipbb)(H(2)hfipbb)(0.5), has unprecedented selectivity for membrane-based separation of CO(2)/CH(4) mixtures. Our calculations combine molecular dynamics, transition state theory, and plane wave DFT calculations to assess the importance of framework flexibility in the MOF during molecular diffusion. This combination of methods should also make it possible to identify other MOFs with attractive properties for kinetic separations. PMID- 20024408 TI - Water structuring and collagen adsorption at hydrophilic and hydrophobic silicon surfaces. AB - The adsorption of a collagen fragment on both a hydrophobic, hydrogen-terminated and a hydrophilic, natively oxidised Si surface is investigated using all-atom molecular dynamics. While favourable direct protein-surface interactions via localised contact points characterise adhesion to the hydrophilic surface, evenly spread surface/molecule contacts and stabilisation of the helical structure occurs upon adsorption on the hydrophobic surface. In the latter case, we find that adhesion is accompanied by a mutual fit between the hydrophilic/hydrophobic pattern within the protein and the layered water structure at the solid/liquid interface, which may provide an additional driving force to the classic hydrophobic effect. PMID- 20024409 TI - Enhanced dihydrogen adsorption in symmetry-lowered metal-porphyrin-containing frameworks. AB - Porphyrin is a very important component of natural and artificial catalysis and oxygen delivery in blood. Here, we report that, based on first-principles density functional calculations, a hydrogen molecule can be adsorbed non-dissociatively onto Ti-, V-, and Fe-porphyrins, similar to oxygen adsorption in heme-containing proteins, with a significant energy gain, greater than 0.3 eV per H(2). The dihydrogen-heme complex will be non-magnetic, as is oxyhemoglobin. In contrast to the backward electron donation of Fe(III)-O(2)(-) in oxyhemoglobin, the dihydrogen binding originates from electron donation from H(2) to the Fe(II). We have identified that the local symmetry of the transition metal center of porphyrins uniquely determines the binding strength, and, thus, one can even manipulate the strength by intentionally and systematically breaking symmetry. PMID- 20024410 TI - NMR tensors in planar hydrocarbons of increasing size. AB - (13)C nuclear shielding and (13)C-(13)C spin-spin coupling tensors were calculated using density functional theory linear response methods for a series of planar hydrocarbons. As calculation of the spin-spin coupling is computationally demanding for large molecules due to demands placed on basis-set quality, novel, compact completeness-optimized (co) basis sets of high quality were employed. To maximize the predictive value of the data, the convergence of the co basis sets was compared to well-known basis-set families. The selection of the exchange-correlation functional was performed based on the available experimental data and coupled-cluster calculations for ethene and benzene. The series of hydrocarbons, benzene, coronene, circumcoronene and circumcircumcoronene, was chosen to simulate increasingly large fragments of carbon nanosheets. It was found that the nuclear shielding and the one-, two-, and three-bond spin-spin coupling constants, as well as the corresponding anisotropies with respect to the direction normal to the plane, approach convergence as the number of carbon atoms in the fragment is increased. Predictions of the investigated properties can then be done for the limit of large planar hydrocarbons or carbon nanosheets. From the results obtained with a judicious choice of the functional, PBE, and co basis close to convergence, limiting values are estimated as follows: sigma = 54 +/- 1 ppm [corresponding to the chemical shift of 134 ppm with methane (CH(4)) as a reference], Deltasigma = 207 +/- 4 ppm, (1)J = 59.0 +/- 0.5 Hz, Delta(1)J = -1.5 +/- 0.5 Hz, (2)J = 0.2 +/ 0.4 Hz, Delta(2)J = -4.6 +/- 0.2 Hz, (3)J = 6 +/- 1 Hz, and Delta(3)J = 3 +/- 1 Hz. PMID- 20024411 TI - Spin-Peierls transition in low-dimensional quantum spin systems: a Green's function approach. AB - We study the spin-Peierls (SP) transition of one-dimensional chain polymeric complexes coupled to lattice by means of many-body Green's function theory. The chain effective elastic constant is an intrinsic factor that determines the order of SP transition. It is found that the SP transition temperature T(SP) and the susceptibility-maximum temperature T(max) are in agreement with the experimental results. When an external magnetic field is applied to the chain, it makes T(SP) and T(max) decrease, and drives the SP transition from the second order to the first order. Besides, we show that the two-site thermal entanglement entropy is a good indicator of SP transition. Further considering the effect of interchain coupling on SP transition, with weak coupling of double-chain, the theoretical values are closer to the experimental results. We also calculate the density of states and spectral functions, which show that the energy gap vanishes at a critical temperature lower than T(SP), indicating a gapless SP phase lies in the gapped dimerized phase. The interchain coupling can drive the SP transition from the second order to the first order, while the SP dimerization may collapse for large interchain couplings. PMID- 20024412 TI - Cooperativity of pi-stacking and hydrogen bonding interactions and substituent effects on X-ben//pyr...H-F complexes. AB - Quantum chemical calculations have been performed to gauge the effect of pi stacking and hydrogen bonding interactions on each other in X-ben//pyr...H-F (X = NO(2), CF(3), CN, F, Cl, CH(3) and OH) complexes. The results indicate the cooperativity of interactions in these complexes where face-to-face aromatic interactions and hydrogen bonding interactions coexist. The effects of substituents on the X-ben//pyr...H-F complexes have also been studied with the MP2 method using 6-31G** basis set. The total binding energy increases in both electron-donating and withdrawing substituents. Herein, computational results indicate an enhanced pi-stacking interaction for all substituted complexes related to an unsubstituted case. On the other hand, H...N hydrogen bond interaction is declined by strong electron withdrawing substituents (NO(2) and CN) only. The cooperativity of pi-stacking and H...N hydrogen bond interaction has also been studied by using the atoms in molecules (AIM), natural bond orbital (NBO) and molecular electrostatic potential (MEP) analyses. There are good relationships between the Hammett constants and energy data, geometrical parameters, and the results of population analysis in X-ben//pyr...H-F and X ben//pyr complexes. The characteristics of interactions are directly related to the electrostatic interaction between the rings. PMID- 20024413 TI - Electronic structure of disjoint diradical 4,4'-bis(1,2,3,5-dithiadiazolyl) thin films. AB - The electronic structure of the thiazyl diradical, 4,4'-bis(1,2,3,5 dithiadiazolyl) (BDTDA), has been investigated by ultraviolet photoemission spectroscopy. Stacked BDTDA dimers showed an energy band dispersion of about 0.3 eV for the highest occupied molecular orbital in the direction of the surface normal of the BDTDA solid film. The pi-orbital overlap between the stacked dimers therefore evolves into a quasi one-dimensional energy band along the dimer stacking direction. PMID- 20024414 TI - Mechanism of electro-oxidation of carbon monoxide on stepped platinum electrodes in alkaline media: a chronoamperometric and kinetic modeling study. AB - From a detailed analysis of the chronoamperometry of CO stripping on stepped platinum single-crystal electrodes in alkaline solution, in combination with kinetic modeling, a mechanistic and kinetic picture of the CO oxidation mechanism is derived. On Pt(111), CO oxidation starts at defect sites (steps, kinks), following a one-dimensional nucleation-and-growth mechanism, or a Langmuir Hinshelwood mechanism with no effective competition between CO and OH. The carbonate that is formed in this reaction blocks the active oxidation sites, so that CO adsorbed on terraces further away from the defect sites can be oxidized at defects sites only very slowly. At potentials above ca. 0.75 V vs. RHE this CO is oxidized on the Pt(111) terrace. On stepped Pt electrodes, CO oxidation is also initiated at the kink (step defects) and step sites. We postulate that carbonate partially blocks the active site, but CO from the terrace still prefers to react at the steps over the terrace sites. The oxidation of terrace-bound CO at the step sites follows a competitive Langmuir-Hinshelwood mechanism. The least reactive CO on the surface is the CO that is adsorbed on the step sites, and it is oxidized by OH on terraces. Finally, on the terrace, the Tafel slope for CO stripping is close to 60 mV dec(-1), suggesting an EC mechanism, i.e. reversible OH formation followed by a chemical rate-determining step, presumably the CO + OH combination reaction. At the step site, the Tafel slope is close to 120 mV dec( 1), suggesting an electrochemical rate-determining step, ascribed to slow OH formation at the step site. PMID- 20024415 TI - Graph-topological approach to magnetic properties of benzenoid hydrocarbons. AB - Application of topological properties and graph theory to benzenoid hydrocarbons allowed us to construct an effective approach interpreting ring current formation in molecules when exposed to an external magnetic field. Transformation of unexcited canonical structures for molecules of 34 benzenoid hydrocarbons into circuit structures and then to directed circuit structures allowed us to define global magnetic characteristics (GMC). GMC/n(2) values correlate very well with exaltation of magnetic susceptibility DeltaLambda/n(2) (computed at the CSGT/B3LYP/6-311G** level of theory by using optimized geometries at the B3LYP/6 311G** DFT level) with cc = 0.993. If the approach is applied to individual rings, then the correlation between local magnetic characteristics (LMC) for 129 various rings of 34 benzenoid hydrocarbons and NICS(1) works with cc = -0.975. PMID- 20024416 TI - A detailed comparison of centrifugal sudden and J-shift estimates of the reactive properties of the N + N2 reaction. AB - An extended comparison of the reactive properties of the N + N(2) exchange reaction calculated on a non-collinear dominant potential energy surface using both a centrifugal sudden and a J-shift quantum method is reported. The choice of carrying out such an investigation for N + N(2) is motivated by the fact that the best available (and currently used for spacecraft re-entry simulations) computed set of kinetic data has been worked out using the low level J-shift approximation though based on exact quantum zero total angular momentum probabilities. The fact that our investigation is carried out for a heavy system and a potential energy surface free of wells in the strong interaction region minimizes the occurrence of tunnel, resonance and interference effects which would make the rationalization of the result difficult and the centrifugal sudden treatment less accurate. The study has provided evidence of two important limits of the J-shift approximation: the wrong determination of the maximum value of the total angular momentum quantum number J contributing to reactivity and the lack of deformation of the partial reactive probability dependence on energy at fixed J value. Accordingly, it has been found that the J-shift state-specific cross sections underestimate the corresponding CS values when the initial diatomic rotational energy is low while the situation reverses when the initial diatomic rotational energy is high. PMID- 20024417 TI - On the stability of X2NO radicals (X = F, Cl, Br, I). AB - The stability of X(2)NO radicals has been studied by investigating the three possible dissociative channels, namely X+XNO, X(2)+NO, and NX(2)+O. While all the radicals have been found stable with respect to the latter, the second pathway shows that Br(2)NO and I(2)NO are unstable with respect to dissociation. The first dissociative channel has been thoroughly investigated with the aim of understanding whether F(2)NO and Cl(2)NO are stable and how much. This implied the molecular structures and energies of X(2)NO and XNO, with X = F, Cl, to be computed at high level of theory. The coupled cluster ansatz in conjunction with hierarchical series of basis sets has been employed, thus accounting for extrapolation to the complete basis-set limit. Core correlation as well as higher excitations in the electronic-correlation treatment have also been taken into account. It is particularly noteworthy that explicit inclusion of quadruple excitations allowed to obtain for the first time equilibrium geometries of FNO and ClNO in full quantitative agreement with their experimental counterparts. PMID- 20024418 TI - Resonance Raman spectra of beta-carotene in solution and in photosystems revisited: an experimental and theoretical study. AB - The resonance Raman (RR) spectra of beta-carotene have been studied in solution and in the protein complexes of photosystems I and II (PS I, PS II). The experimental studies are complemented by density functional theory (DFT) calculations that allow for a consistent assignment of most of the experimental RR bands in the region between 900 and 1650 cm(-1). Thus, it was shown that the prominent peak at ca. 1525 cm(-1) is composed of two closely spaced modes, both of which dominated by C=C stretching coordinates of the polyene chain. These two modes exhibit different excitation profiles in the region of the allowed electronic S(0) --> S(2) transition with maxima that are separated by ca. 1300 cm(-1). Calculated RR spectra and excitation profiles obtained by time-dependent DFT in conjunction with the transform method indicate that the enhancement pattern of these modes cannot be rationalised within the Condon approximation (A term scattering). Furthermore, symmetry considerations rule out enhancement via vibronic coupling. Instead, we suggest that the different excitation profiles of the two modes result from force constant changes and mode mixing upon electronic transition (Dushinsky rotation). RR spectra and excitation profiles are very similar in solution and in the protein complexes. Thus, the excitation-dependent frequency variation of the 1525-cm(-1) peak in PS I and PS II is an intrinsic molecular property of beta-carotene and does not reflect different pools of the pigments in specific molecular environments. PMID- 20024419 TI - Electronically excited states of protonated aromatic molecules: benzaldehyde. AB - The photofragmentation spectrum of protonated benzaldehyde has been recorded in the 435-385 nm wavelength range. The first excited state is a pipi* state, strongly red shifted compared to the pipi* state of neutral benzaldehyde. The spectrum presents well resolved vibronic bands in contrast to some other protonated aromatic molecules like benzene or tryptophan in which the excited state dynamics is so fast that no vibrational structure can be observed. The bands can be assigned on the basis of a Franck-Condon analysis using ground and excited state frequencies calculated at the CC2/TZVP level. PMID- 20024420 TI - Mg-25 ultra-high field solid state NMR spectroscopy and first principles calculations of magnesium compounds. AB - Due to sensitivity problems, (25)Mg remains a largely under-explored nucleus in solid state NMR spectroscopy. In this work at an ultrahigh magnetic field of 21.1 T, we have studied at natural abundance the (25)Mg solid state (SS) NMR spectra for a number of previously unreported magnesium compounds with known crystal structures. Some previously reported compounds have been revisited to clarify the spectra that were obtained at lower fields and were either not sufficiently resolved, or misinterpreted. First principles calculations of the (25)Mg SS NMR parameters have been carried out using plane wave basis sets and periodic boundary conditions (CASTEP) and the results are compared with experimental data. The calculations produce the (25)Mg absolute shielding scale and give us insight into the relationship between the NMR and structural parameters. At 21.1 T the effects of the quadrupolar interactions are reduced significantly and the sensitivity and accuracy in determining chemicals shifts and quadrupole coupling parameters improve dramatically. Although T(1) measurements were not performed explicitly, these proved to be longer than assumed in much of the previously reported work. We demonstrate that the chemical shift range of magnesium in diamagnetic compounds may approach 200 ppm. Most commonly, however, the observed shifts are between -15 and +25 ppm. Quadrupolar effects dominate the (25)Mg spectra of magnesium cations in non-cubic environments. The chemical shift anisotropy appears to be rather small and only in a few cases could the contribution of the CSA be detected reliably. A good correspondence between the calculated shielding constants and experimental chemical shifts was obtained, demonstrating the good potential of computational methods in spectroscopic assignments of solid state (25)Mg NMR spectroscopy. PMID- 20024421 TI - Diffusion coefficients and local structure in basic molten fluorides: in situ NMR measurements and molecular dynamics simulations. AB - The local structure and the dynamics of molten LiF-KF mixtures have been studied by nuclear magnetic resonance (NMR) and molecular dynamics simulations. We have measured and calculated the self-diffusion coefficients of fluorine, lithium and potassium across the full composition range around the liquidus temperature and at 1123 K. Close to the liquidus temperature, D(F), D(Li) and D(K) change with composition in a way that mimics the phase diagram shape. At 1123 K D(F), D(Li) and D(K) depend linearly on the LiF molar fraction. These results show that the composition affects the self-diffusion of anions and cations more weakly than the temperature. The activation energy for diffusion was also determined and its value can be correlated with the strength of the anion-cation interaction in molten fluoride salts. PMID- 20024422 TI - X-ray reflectometry studies on the effect of water on the surface structure of [C4mpyr][NTf2] ionic liquid. AB - The effect of water on the surface structure of 1-butyl-1-methylpyrrolidinium trifluoromethylsulfonylimide [C(4)mpyr][NTf(2)] ionic liquid was investigated using X-ray reflectometry. The measured reflectivity data suggests a significant amount of water is adsorbed at the surface, with the first layer from the gas (nitrogen)-liquid phase boundary mainly occupied by a mixture of cations and water. Beyond the cation + water layer, the scattering length density increases towards the bulk value, indicating a decreasing amount of water and cations, and/or an increasing amount of anions. The orientation of the butyl chain of cation at the phase boundary and the population of water at the surface were described based on results from an independent molecular dynamics (MD) simulation. We show that the presence of water in the ionic liquid has a non monotonic effect on the overall thickness of the surface. At low water content, the addition of water does not change the surface thickness since water is mainly present in the bulk. As the water content increases, the surface swells before eventually shrinking down close to the solubility limit of water. The non monotonic surface thickness is used to explain the anomalous trend of surface tension in ionic liquid-water mixtures reported in the literature. PMID- 20024423 TI - Amplified spontaneous emission from opal photonic crystals engineered with structural defects. AB - In this work, we report on the optical properties and amplified spontaneous emissions (ASE) of polystyrene opals engineered with planar structural defects containing a conjugated polymer emitter. Defects in opals give rise to allowed states inside the photonic stop band, which are probed by transmittance and reflectance spectroscopy. The emission spectrum of the polymer embedded in the defect layer is strongly modified and fingerprints of defect states located inside the stop band are recognized. Amplified spontaneous emission for these engineered photonic crystals is clearly observed. PMID- 20024424 TI - The OH + D2 --> HOD + D angle-velocity distribution: quasi-classical trajectory calculations on the YZCL2 and WSLFH potential energy surfaces and comparison with experiments at ET = 0.28 eV. AB - The angle-velocity distribution (HOD) of the OH + D(2) reaction at a relative translational energy of 0.28 eV has been calculated using the quasi-classical trajectory (QCT) method on the two most recent potential energy surfaces available (YZCL2 and WSLFH PESs), widely extending a previous investigation of our group. Comparison with the high resolution experiments of Davis and co workers (Science, 2000, 290, 958) shows that the structures (peaks) found in the relative translational energy distributions of products could not be satisfactorily reproduced in the calculations, probably due to the classical nature of the QCT method and the importance of quantum effects. The calculations, however, worked quite well for other properties. Overall, both surfaces led to similar results, although the YZCL2 surface is more accurate to describe the H(3)O PES, as derived from comparison with high level ab initio results. The differences observed in the QCT calculations were interpreted considering the somewhat larger anisotropy of the YZCL2 PES when compared with the WSLFH PES. PMID- 20024425 TI - Low temperature behavior of thermodynamic perturbation theory. AB - Monte Carlo simulations have been carried out for a hard sphere square well model fluid with well widths of lambda = 1.01, 1.02 and 1.04 to obtain thermodynamic properties, such as pressure, excess Helmholtz free energy and internal energy, constant volume excess heat capacity, excess chemical potential, and excess enthalpy. The extremely narrow well widths considered allowed us to perform simulations at extremely low temperatures without reaching the liquid-vapour transition, which is metastable for these values of lambda. These simulation data have been used to explore the low temperature behavior of two thermodynamic perturbation theories (TPT), namely a high temperature series expansion truncated at second order within the local compressibility approximation (thereafter denoted as 2nd-order LCA-TPT) and a recently proposed coupling parameter series expansion truncated at 3rd-, 4th- and 5th-order (thereafter denoted as 3rd-order, 4th-order and 5th-order TPT, respectively). It has been found that the 2nd-order LCA-TPT is qualitatively incorrect in most of the cases analyzed, whereas the 3rd order TPT is quantitatively correct in most of them. With increasing the well width, and consequently the temperatures considered, the 3rd-order TPT quickly becomes more accurate. Among the six thermodynamic quantities analyzed, the one most difficult to predict accurately is the constant volume excess heat capacity, for which the TPT based on the coupling parameter expansion provides satisfactory results only in the case of lambda = 1.04. For the cases studied, the performance of the 3rd-order, 4th-order, and 5th-order TPT are essentially equal; the change of the order of the truncation sometimes results in an improvement of some thermodynamic quantities within certain regions of the thermodynamic surface of states, while worsening in others. PMID- 20024426 TI - Theoretical study of solvent effect on one- and two-photon absorption properties of starburst DCM derivatives. AB - The equilibrium geometries, electronic structures, one-photon absorption (OPA) and two-photon absorption (TPA) properties of starburst 4-(dicyanomethylene)-2 methyl-6-[p-(dimethylamino)styryl]-4H-pyran (DCM) derivatives have been studied by using density functional theory (DFT) and Zerner's intermediate neglect of differential overlap (ZINDO) program. Results showed that increasing the number of branches in DCM molecules and introducing stronger electron withdrawing groups are efficient ways to enhance the TPA cross sections. For the two- and three branched molecules of the DCM derivatives, the solvent effect on the OPA and TPA properties was also considered by adapting the self-consistent reaction field (SCRF) theory with the polarizable continuum model (PCM) to model the interaction with the solvent in ZINDO program. The calculated molecular structure, transition dipole moment and intramolecular charge transfer (ICT) in different solvents have been analyzed to approximate how the solvent affects the TPA cross section. Results indicated that the TPA cross section increases as the polarity of the solvents increases, and it reaches a maximum when the dielectric constant epsilon equals 20.7. PMID- 20024427 TI - A DFT comparative study of carbon adsorption and diffusion on the surface and subsurface of Ni and Ni3Pd alloy. AB - Carbon diffusion in transition metal nanoparticles is assumed to be a key factor in the catalyzed growth of carbon nanotubes (CNT). Aiming at designing more efficient catalysts, we have compared this carbon diffusion process in the near surface and in the bulk of Ni and Ni(3)Pd by means of density functional theory (DFT) calculations. Ni nanoparticles are indeed the most largely used catalysts and the alloying with Pd could modify and improve their properties. The alloy has the same crystal structure as pure Ni, with a slight lattice expansion due to the presence of palladium. For both systems, the subsurface octahedral site is the most stable adsorption site, but the thermodynamic trend favoring the penetration to the subsurface is larger on the alloy than on the Ni. As a result, in the conditions of temperature and pressure for nanotube growth, the population of the subsurface sites is a more exothermic process on the alloy. In addition, while on pure nickel the diffusion over the (111) surface is easy, on the alloy the vertical process leading the carbon to the subsurface is preferred. Palladium atoms have the double effect to expand the lattice parameter providing more adapted diffusion channels for the carbon and to create new adsorption sites less stable than the all-nickel ones. The results can be related to more selective formation of nanotubes on the alloy at low temperature, where Ni produces fibers. PMID- 20024428 TI - Kinetic model of the electrochemical oxidation of graphitic carbon in acidic environments. AB - The electrochemical oxidation of graphitic carbon results in the performance decay of electrochemical systems such as aqueous, acidic fuel cells, redox-flow batteries, and supercapacitors. An electrochemical mechanism and numerical model is proposed to explain long-standing questions. The model predicts carbon weight loss and surface oxide growth as a function of time, temperature, and potential. Experimentally observed phenomena are discussed and analyzed using the numerical model. Three mechanisms are concluded to contribute to the current decay commonly observed during electrochemical oxidation: mass loss, reversible passive oxide formation, and irreversible oxide formation. Although reversible passive oxide formation governs the current decay under potentiostatic oxidation, a reduction in the equilibrium catalytic oxide is the most significant decay mechanism under potential cycling. Finally, the model is used to determine the change in active site concentration resulting from high-temperature heat treatment of carbon black. PMID- 20024429 TI - Re-dissolution and de-compaction of DNA-cationic surfactant complexes using non ionic surfactants. AB - Addition of a cationic surfactant to a solution of DNA causes the formation of compacted DNA-cationic surfactant complexes which precipitate from aqueous solution. It has been shown previously that addition of anionic surfactant will re-dissolve and de-compact the DNA-cationic surfactant complexes and we find that addition of non-ionic surfactants of the alkylpolyoxyethylene type can be used similarly. In principle, these de-compaction and re-dissolution processes could occur either by stripping of the cationic surfactant from the DNA into mixed micelles with the non-ionic surfactant or by solubilisation of the DNA-cationic surfactant complexes within the non-ionic micelles. Solubility phase-boundary measurements, fluorescence microscopy observations of the de-compaction process and light scattering results indicate that de-compaction and re-dissolution occur by the stripping mechanism, even for non-ionic surfactants where the favourable attractive electrostatic interaction between the two surfactants is absent. Using measurements of critical micelle concentrations and calculations based on regular solution mixed micelle theory, we show that re-dissolution and de-compaction of the DNA-cationic surfactant complexes occurs when the concentration of free monomeric cationic surfactant is reduced (by incorporation into mixed micelles) below a critical value. PMID- 20024430 TI - Influence of salt on assembly and compression of PDADMAC/PSSMA polyelectrolyte multilayers. AB - Influence of NaCl and NaBr and their concentration on assembly of polyelectrolyte multilayers and their compression behavior was studied using poly(diallyldimethylammonium chloride) (PDADMAC) and poly(4-styrenesulfonic acid co-maleic acid, 1 : 1 SS:MA) sodium salt (PSSMA 1 : 1) as the building blocks. Stepwise growth of the PSSMA/PDADMAC multilayers was characterized by a quartz crystal microbalance with dissipation (QCM-D), revealing faster and slower growth of the layer thickness in higher concentration of NaCl and NaBr solutions, respectively. Moreover, QCM-D, ellipsometry and scanning force microscopy (SFM) measurements demonstrated that the multilayers were highly hydrated with a factor of 3-9 in a wet state depending on the type and concentration of the salts used during the assembly. The type and concentration of the salts used during the assembly also influenced greatly the compression ratio of the polyelectrolyte multilayers, i.e. the compression ratio increased along with the increase of NaCl concentration, whereas it was constant when NaBr was used. The friction force of the PEI(PSSMA/PDADMAC)(7) multilayers measured by SFM showed that the force peaks gradually shifted from low to high scanning frequency along with the increase of NaCl concentration, illustrating the gradual softening of the multilayers. By contrast, the peak positions in the friction force curves for all the multilayers assembled in NaBr solution kept unchanged regardless of the NaBr concentration. Contrast experiments further revealed that the compression ratios, masses in wet and wet thicknesses of the multilayers alternatingly treated in Cl(-) and in Br( ) solutions were all reversible. PMID- 20024431 TI - Integral and differential cross sections for the S(1D)+HD reaction employing the ground adiabatic electronic state. AB - We present converged quantum mechanical calculations for the title reaction employing a time-dependent wavepacket method. We obtained integral and differential cross sections over an energy range from 0.23 to 0.35 eV total energy as well as product state distributions for both product channels. The excitation functions decrease with energy and point to statistical dynamics as do the cold vibrational distributions and highly inverted rotational distributions. The differential cross sections oscillate strongly with energy for both product channels. Our differential cross sections for both product channels at 2.5 kcal/mol, one of the experimental energies, compare well to the experimental results. The quantum results obtained in this study are similar to what has been found employing QCT methods, implying that the differences between the experimental and theoretical results are due to the potential energy surface or non-adiabatic effects rather than due to quantum effects or the methods employed. PMID- 20024432 TI - Impact of vibrational excitation on the kinetics of a nascent ketene. AB - The formation and decay of a ketene intermediate photochemically formed from o nitrobenzaldehyde has been studied by femtosecond UV/Vis and IR spectroscopy. The ketene is formed predominantly within a few 100 fs and to a minor extent within approximately 200 ps via the recombination of a triplet phased bi-radical. In tetrahydrofuran solution the ketene intermediate is seen to form a secondary intermediate with biphasic kinetics. The first phase of this decay occurs within a few picoseconds. It can be attributed to the reaction of vibrationally excited ketenes. The second phase characterized by a time constant of 2 ns is due to the reaction of thermalized molecules. In 2-butanol solution the lifetime of the thermalized ketene is only approximately 60 ps and the rapid and the slow phases of the decay start to merge. PMID- 20024433 TI - Molecular recognition in the gas phase: benzocaine-phenol as a model of anaesthetic-receptor interaction. AB - The benzocaine-phenol complex is proposed as a model system of the interaction between the local anaesthetic benzocaine and the tyrosine residue. The complex has been generated by supersonic expansion of benzocaine and phenol in helium and probed by 1- and 2-color mass-resolved laser spectroscopies. The electronic excitation spectrum of the 1 : 1 complex spans some approximately 700 cm(-1) and includes well resolved bands from at least two isomers, as demonstrated using UV UV hole burning spectroscopy. The combination of ion dip infrared spectroscopy (IDIRS) and ab initio calculations shows that both isomers are stabilized by an OH...N hydrogen bond between the phenol hydroxyl group and the benzocaine amino moiety, differing only in the conformation adopted by the benzocaine monomer (trans and gauche). The application of the fragmentation threshold method to benzocaine-phenol suggests the existence of chemical reactions in the electronic excited state of the complex and/or in the ion. Such hypothesis is also supported by the calculated potential energy curves along the hydrogen bond coordinate. PMID- 20024434 TI - Non covalent interactions in RNA and DNA base pairs: a quantum-mechanical study of the coupling between solvent and electronic density. AB - It is well-known that a solvent can modify the relative importance of the different constituents (electrostatic and dispersion) of non-covalent interactions, but much less is known about how these solvent-induced modifications specifically couple with the polarization of the electronic density and electronic correlation. Here we present a quantum mechanical analysis of the effects of the solvent on the non covalent interactions (both stacking and hydrogen bonding) in base pairs using a hierarchy of combinations between a MP2 correlated description for the base pairs and the polarizable continuum model (PCM) for the solvent. A comparison of the results obtained in these different combinations of increasing accuracy allows us to better analyze the important role played by the coupling between correlated electronic densities and solvent polarization in determining the relative importance of stacking and hydrogen bonding effects. PMID- 20024435 TI - A multi-technique approach to the analysis of SAMs of aromatic thiols on copper. AB - The adsorption of aromatic thiols on Cu and the SAM film stability in acidic solutions have been studied by XPS, contact angle and electrochemical techniques. Three short molecules, benzenethiol (BT), 2-naphthalenethiol (2-NT) and 4 acetamidothiophenol (4-AA), were selected as representatives of aromatic thiols to highlight the effect of aromatic rings and hydrophilic terminal groups on the copper protection. All the three molecules form stable S-Cu bonds as a consequence of their adsorption process on polycrystalline copper. Although none of them provides a full copper passivation, the adsorbed films persist without major degradation on Cu electrodes even after 12 h immersion in 0.5 M sulfuric acid. Comparing the freshly prepared adsorbed films, the larger 2-NT molecule provides a better Cu passivation, but the shorter BT molecule favours a higher surface coverage. The terminal groups of 4-AA are responsible for a higher Cu surface wettability in water, compared to that with SAMs of the other molecules, and allow for an easier charge-transfer to the electrolyte and for a higher electrochemical capacitance. After long enough ageing, however, the 4-AA-based molecular films are able to self-organize and to provide a steadily improving copper passivation. Adlayers of the BT and 2-NT molecules, on the contrary, over a long time tend to protect less and less the Cu substrate, probably because of progressive electrolyte infiltration. PMID- 20024436 TI - Electron attachment and electron ionization of acetic acid clusters embedded in helium nanodroplets. AB - The effect of incident electrons on acetic acid clusters is explored for the first time. The acetic acid clusters are formed inside liquid helium nanodroplets and both cationic and anionic products ejected into the gas phase are detected by mass spectrometry. The cation chemistry (induced by electron ionization at 100 eV) is dominated by production of protonated acetic acid (Ac) clusters, Ac(n)H(+), although some fragmentation is also observed. In the case of anion production (at 2.8 eV electron energy) there is a clear distinction between the monomer and the clusters. For the monomer the dominant product is the dehydrogenated species, [Ac-H](-), whereas for the clusters both the parent anion, Ac(n)(-), and the dehydrogenated species, [Ac(n)-H](-), have similar abundances. A particularly intriguing contrast between the monomer and cluster anions is that helium atoms are seen attached to the latter whereas no evidence of helium atom attachment is found for the monomer. This surprising observation is attributed to the formation of acyclic (head-to-tail) acetic acid clusters in helium nanodroplets, which have more favourable electronic properties for binding helium atoms. The acyclic clusters represent a local minimum on the potential energy surface and in the case of the dimer this is distinct from the cyclic isomer (the global minimum) identified in gas phase experiments. PMID- 20024437 TI - Principal component analysis of potential energy surfaces of large clusters: allowing the practical calculation of the master equation. AB - The number of variables in many-particle systems is typically unmanageably large; some way to reduce that number and still retain access to the important information about the system of interest is one of the great challenges in the broad topic of complexity. Principal components and principal coordinates provide a powerful means to extract--from unwieldy, large data sets--a reduced collection of variables that provide the information one needs, in a relatively efficient way and useful form. We investigate the application of principal components to the analysis of kinetics of the atomic motions in atomic clusters, particularly of clusters that are large enough so that a full description in terms of the entire high-dimensional potential surface is entirely impractical. A specific application is the use of principal components linking minima with their adjacent saddles, permitting the evaluation of rate coefficients (in the context of transition state theory) as ratios of partition functions of only one or two key variables. PMID- 20024438 TI - XPS study of ruthenium tris-bipyridine electrografted from diazonium salt derivative on microcrystalline boron doped diamond. AB - Boron doped diamond (BDD) functionalization has received an increasing interest during the last few years. Such an infatuation comes from the original properties of BDD, including chemical stability or an electrochemical window, that opens the way for the design of (bio)sensors or smart interfaces. In such a context, diazonium salts appear to be well suited for BDD functionalization as they enable covalent immobilization of functional entities such as enzymes or DNA. In this study we report microcrystalline BDD functionalization with a metallic complex, ruthenium tris(bipyridine), using the p-(tris(bipyridine)Ru(2+))phenyl diazonium salt. Electrografting using cyclic voltammetry (CV) allowed the formation of a ruthenium complex film that was finely characterized using electrochemistry and X ray photoelectron spectroscopy (XPS). Moreover, we showed that chronopotentiometry (CP) is a convenient tool to monitor Ru complex film deposition through the control of the electrochemical pulse parameters (i.e. current density and pulse duration). Finally, such a control was demonstrated through the correlation between electrochemical and XPS characterizations. PMID- 20024440 TI - Interaction of internally mixed aerosols with light. AB - Atmospheric aerosols scatter and absorb solar radiation leading to variable effects on Earth's radiative balance. Aerosols individually comprising mixtures of different components ("internally mixed") interact differently with light than mixtures of aerosols, each comprising a different single component ("externally mixed"), even if the relative fractions of the different components are equal. In climate models, the optical properties of internally mixed aerosols are generally calculated by using electromagnetic "mixing rules", which average the refractive indices of the individual components in different proportions, or by using coated sphere Mie scattering codes, which solve the full light scattering problem assuming that the components are divided into two distinct layers. Because these calculation approaches are in common use, it is important to validate them experimentally. In this article, we present a broad perspective on the optical properties of internally mixed aerosols based on a series of laboratory experiments and theoretical calculations. The optical properties of homogenously mixed aerosols comprised of non-absorbing and weakly absorbing compounds, and of coated aerosols comprised of strongly absorbing, non-absorbing, and weakly absorbing compounds in different combinations are measured using pulsed and continuous wave cavity ring down aerosol spectrometry (CRD-AS). The success of electromagnetic mixing rules and Mie scattering codes in reproducing the measured aerosol extinction values is discussed. PMID- 20024442 TI - Supramolecular nanoarchitectures for light energy conversion. AB - Recent developments in synthetic and supramolecular techniques have made it possible to control precisely, organize and arrange molecules at the nanometre level. Such synthetic and supramolecular strategies enable us to construct photofunctional molecular architectures for light energy conversion, such as photovoltaics. In photovoltaic cells, processes such as light-harvesting, charge separation for carrier generation, and carrier transport are generally required. Therefore, the construction of supramolecular assemblies based on these three processes is interesting and promising for the future development of photovoltaics. In this perspective, the focus is on the recent developments of supramolecular systems for light energy conversion, which are mainly composed of porphyrin dyes and nanocarbon materials, such as fullerenes and carbon nanotubes. The specific topics are as follows: (i) preparation, photodynamics, and photoelectrochemistry of self-assembled porphyrin nanoparticles prepared by simple blend, (ii) highly organized supramolecular nanoassemblies of porphyrins and fullerenes using gold nanoparticles, dendritic and polypeptide structures, (iii) the supramolecular formation and photoelectrochemical property of carbon nanotubes, and (iv) supramolecular photofunctional nanorods of porphyrins. PMID- 20024441 TI - Photoinduced short-range electron transfer in DNA with fluorescent DNA bases: lessons from ethidium and thiazole orange as charge donors. AB - Charge transfer processes through the double helix of DNA cover a broad range of mechanistic models ranging from superexchange to hopping mechanisms. Over the last decade, these processes were studied by our group in a photoinduced fashion since (i) the starting time for the charge transfer is clearly defined by the absorption of the photon and (ii) photoexcitation delivers the necessary driving force to the DNA system. It is a prerequisite to modify oligonucleotides synthetically with suitable organic fluorophores that serve as photoinducable charge donors. In the first part of this perspective article we summarize our recent advances in the area of DNA-mediated reductive electron transfer processes over short ranges using synthetic DNA-donor-acceptor systems. The second part of this article focuses on ethidium as the photoinducable charge donor. Ethidium modified DNA can be used to compare oxidative hole transfer with reductive electron transfer since the type of charge transfer can be controlled by choosing the right charge acceptor. Recent results showed that an efficient charge transfer through DNA using covalently bound ethidium is strongly influenced mainly by DNA dynamics but also by several other parameters that affect the electronic coupling between charge donor and acceptor. PMID- 20024443 TI - A bifunctional strategy towards experimentally (synthetically) attainable molecules with planar tetracoordinate carbons. AB - Bifunctional strategy, including efficiently utilizing valence electrons and offering steric protection, has been proposed to advance C(2)Al(4) global minimum with double planar tetracoordinate carbons (ptC) to a new family of ptC molecules which could be promising for synthetic realization. PMID- 20024444 TI - Resolving ligand hyperfine couplings of type 1 and 2 Cu(II) in ascorbate oxidase by high field pulse EPR correlation spectroscopy. AB - Ascorbate oxidase contains two paramagnetic Cu(ii) binding sites, type 1 (T1) and type 2 (T2) and in both sites the Cu(ii) is coordinated to histidine residues. We use several pulse EPR techniques at high field (95 GHz) to determine ligand (1)H and (14)N hyperfine couplings in the two sites and identify the T1 signals by a new triple resonance correlation technique named THYCOS. PMID- 20024445 TI - On the efficiency limit of triplet-triplet annihilation for photochemical upconversion. AB - Photochemical upconversion is performed, whereby emitter triplet states are produced through triplet energy transfer from sensitizer molecules excited with low energy photons. The triplet emitter molecules undergo triplet-triplet annihilation to yield excited singlet states which emit upconverted fluorescence. Experiments comparing the 560 nm prompt fluorescence when rubrene emitter molecules are excited directly, using 525 nm laser pulses, to the delayed, upconverted fluorescence when the porphyrin sensitizer molecules are excited with 670 nm laser pulses reveal annihilation efficiencies to produce excited singlet emitters in excess of 20%. Conservative measurements reveal a 25% annihilation efficiency, while a direct comparison between the prompt and delayed fluorescence yield suggests a value as high as 33%. Due to fluorescence quenching, the photon upconversion efficiencies are lower, at 16%. PMID- 20024446 TI - Pseudorotation in pyrrolidine: rotational coherence spectroscopy and ab initio calculations of a large amplitude intramolecular motion. AB - Pseudorotation in the pyrrolidine molecule was studied by means of femtosecond degenerate four-wave mixing spectroscopy both in the gas cell at room temperature and under supersonic expansion. The experimental observations were reproduced by a fitted simulation based on a one-dimensional model for pseudorotation. Of the two conformers, axial and equatorial, the latter was found to be stabilized by about 29 +/- 10 cm(-1) relative to the former one. The barrier for pseudorotation was determined to be 220 +/- 20 cm(-1). In addition, quantum chemical calculations of the pseudorotational path of pyrrolidine were performed using the synchronous transit-guided quasi-Newton method at the MP2 and B3LYP levels of theory. Subsequent CCSD(T) calculations yield the energy preference of the equatorial conformer and the barrier for pseudorotation to be 17 and 284 cm(-1), respectively. PMID- 20024447 TI - The 1,4-phenylenediisocyanide dimer: gas-phase properties and insights into organic self-assembled monolayers. AB - The 1,4-phenylenediisocyanide (PDI) dimer serves as an intriguing case of the substituted benzene dimer, as well as a prototype system for self-assembled monolayers of organic isocyanide complexes. Structures and binding energies are explored using recently developed dual-basis second-order Moller-Plesset perturbation theory energies and gradients. The structures are dictated by a combination of dispersion and electrostatics, a combination not properly treated with local or gradient-corrected density functionals. The PDI dimer binds more than twice as strongly as unsubstituted benzene dimers in several configurations, and greater directional specificity between parallel-displaced and T-shaped structures is observed. A rotated-parallel structure is the predicted lowest energy, gas-phase configuration, in which the isocyanide ligands are staggered on the monomers. Relevant potential energy curves of the dimer are also presented, and insights into PDI monolayer formation on metal surfaces are explored via simple two-body models. Based on the adsorbate interaction alone, a high-coverage configuration and non-vertical tilt are predicted to be favorable, although the total binding for PDI in these configurations is still insufficient to form ordered monolayers, a result consistent with previous experimental findings. Additional phenyl rings (biphenyldiisocyanide, triphenyldiisocyanide) significantly stabilize the interaction and provide the additional dispersion necessary for an ordered monolayer. PMID- 20024448 TI - A UV-Vis micro-spectroscopic study to rationalize the influence of Cl(-)(aq) on the formation of different Pd macro-distributions on gamma-Al(2)O(3) catalyst bodies. AB - The influence of the Cl(-)(aq) concentration, solution pH and equilibration time on the PdCl(4)(2-)(aq) dynamics and molecular structure after impregnation of gamma-Al(2)O(3) catalyst bodies has been studied using UV-Vis micro-spectroscopy. To do so, 0.2 wt% Pd catalysts have been prepared from acidic solutions (pH 1 and 5) of the Na(2)PdCl(4) precursor salt with different amounts of NaCl. It was found that egg-shell catalysts are obtained when a less acidic pH (pH 5) is combined with [Cl(-)(aq)] < 0.6 M and less than 24 h of equilibration time are implemented, while to achieve egg-white catalysts the solution pH should be 1. Moreover, by increasing the equilibration time up to 96 h, the egg-shell profiles vanish to provide a uniform Pd distribution, while the egg-white distribution becomes egg-yolk. Additionally, Pd complexes appeared with different molecular structures depending on the solution pH, equilibration time and macro distribution achieved. The protocol developed to create different Pd macro distributions has been applied to prepare two 1 wt% Pd/gamma-Al(2)O(3) egg-shell and egg-white catalysts. The Pd dynamics and molecular structure have been followed after impregnation, drying and calcination, demonstrating that the profiles created after impregnation are retained. PMID- 20024449 TI - Interplay of slow bath fluctuations and energy transfer in 2D spectroscopy of the FMO light-harvesting complex: benchmarking of simulation protocols. AB - Recently developed approaches to simulate environment-induced fluctuation effects in two-dimensional (2D) spectroscopy of excitons are compared for the Fenna Matthews-Olson light-harvesting complex. Fast fluctuations induce population redistribution between exciton energy-levels and raise homogeneous line widths of various peaks in 2D spectra. These effects are easily accounted for in sum-over eigenstates (SOS) approach and the quasi-particle (QP) scattering approach through relaxation and dephasing rate constants. Slow fluctuations cause correlations of energies at various delay times in 2D photon-echo spectra. These may be calculated either by doing cumulant expansion in SOS techniques or by statistical averaging over static disorder in SOS and QP approaches. We compare the 2D photon-echo signal simulated using two levels of cumulant expansion approaches and two statistical averaging approaches for the same system. These levels differ by the treatment of energy-level correlations at different delay times and give rise to different cross-peak shapes: the cross-peaks retain their original diagonally elongated shapes when correlations are included, while they are more spherically broadened when correlations are neglected. Statistical averaging over disorder give very similar results but requires much higher computational effort. The peak redistribution timescales are very similar for all levels of theory. The spectral signatures at these different levels of theory are compared and simulation cost is estimated. Approaches which do require statistical averaging over disorder are orders of magnitude slower. PMID- 20024450 TI - Quasi-classical statistico-dynamical description of polyatomic photo dissociations: state-resolved distributions. AB - An alternative methodology to investigate indirect polyatomic processes with quasi-classical trajectories is proposed, which effectively avoids any binning or weighting procedure while provides rovibrational resolution. Initial classical states are started in terms of angle-action variables to closely match the quantum experimental conditions and later transformed into Cartesian coordinates, following an algorithm very recently published [J. Chem. Phys. 2009, 130, 114103]. Trajectories are then propagated using the 'association' picture, i.e. an inverse dynamics simulation in the spirit of the exit-channel corrected phase space theory of Hamilton and Brumer [J. Chem. Phys. 1985, 82, 595], which is shown to be particularly convenient. Finally, an approximate quasi-classical formula is provided which under general conditions can be used to add possible rotational structures into the vibrationally-resolved quasi-classical distributions. To introduce the method and illustrate its capabilities, correlated translational energy distributions from recent experiments in the photo-dissociation of ketene at 308 nm [J. Chem. Phys. 2006, 124, 014303] are investigated. Quite generally, the overall theoretical algorithm reduces the total number of trajectories to integrate and allows for fully theoretical predictions of experiments on polyatomics. PMID- 20024451 TI - Endohedral metallofullerenes in self-assembled monolayers. AB - A method has been developed for the attachment of a dithiolane group to endohedral metallofullerenes via a 1,3-dipolar cycloaddition reaction. This sulfur-containing functional group serves as an anchor, enabling efficient immobilisation of endohedral fullerenes on Au(111) surfaces at room temperature, directly from the solution phase. The functionalised fullerenes form disordered monolayers that exhibit no long-range ordering, which is attributed to both the strong bonding of the dithiolane anchor to the surface and to the conformational flexibility of the functional group. Endohedral fullerenes Er(3)N@C(80) and Sc(3)N@C(80) have been used as models for functionalisation and subsequent surface deposition. Their chemical reactivity towards dithiolane functionalisation and their surface behaviour have been compared to that of C(60). The endohedral fullerenes appear to be significantly less reactive towards the functionalisation than C(60), however they bind in a similar manner to a gold surface as their dithiolane terminated C(60) counterparts. The optical activity of Er(3)N@C(80) molecules is preserved after attachment of the functional group. We report a splitting of the endohedral Er(3+) emission lines due to the reduction in symmetry of the functionalised fullerene cage, as compared to the highly symmetrical icosahedral C(80) cage of pristine Er(3)N@C(80). PMID- 20024452 TI - The role of the methyl group in stabilising the weak N-H...pi hydrogen bond in the 4-fluorotoluene-ammonia complex. AB - The 4-fluorotoluene-ammonia van der Waals complex has been studied using a combination of resonant two-photon ionisation (R2PI) spectroscopy, ab initio molecular orbital calculations and multidimensional Franck-Condon analysis. The R2PI spectrum shows two sets of features assignable to two distinct conformers: one in which the ammonia binds between the hydrogen meta to the methyl group and the fluorine atom in a planar configuration and the other a pi-bound structure involving one bond between an ammonia hydrogen and the pi-system and another between the ammonia lone pair and the slightly acidic hydrogens on the methyl group. Ground state estimated CCSD(T) interaction energies were computed at the basis-set limit: these calculations yielded very similar interaction energies for the two conformers, whilst zero point energy correction yielded a zero point binding energy for the pi-complex about 10% larger than that of the in-plane, sigma-complex. The results of multidimensional Franck-Condon simulations based on ab initio ground and excited state geometry optimisations and vibrational frequency calculations showed good agreement with experiment, with further improvements achieved using a fitting procedure. The observation of a pi-complex in addition to a sigma-complex supports the intuitive expectation that electron donating groups should help to increase pi-density and hence stabilise pi-proton acceptor complex formation. In this case, this occurs in spite of the presence of a strongly electron-withdrawing fluorine atom. PMID- 20024453 TI - A sublattice-model isotherm for the competitive coadsorption of hydrogen and bromide on a Pt(100) electrode. AB - Previous work demonstrated that the Frumkin isotherm is inadequate to model the competitive coadsorption of species with different saturation coverages, such as hydrogen and bromide coadsorption on Pt(100) [N. Garcia-Araez et al., J. Electroanal. Chem., 2006, 588, 1]. Therefore, Monte Carlo simulations were necessary to determine meaningful values of the microscopic parameters (namely, energies of adsorption and interaction). In the present work, an alternative analytical isotherm is developed, by taking into account the occupation of two sublattices, which together compose the whole lattice of adsorption sites. Despite its relatively simple mathematical form, this isotherm presents, under certain conditions, a significant improvement over the classical Frumkin isotherm for the modeling of competitive adsorption processes, thus providing a closer agreement with results from Monte Carlo simulations. Finally, it is demonstrated that the sublattice-model isotherm will be generally applicable to systems in which the formation of segregated adlayers, whose structure is not explicitly taken into account in the model, is energetically unfavorable. PMID- 20024454 TI - Theoretical study of H(2) splitting and storage by boron-nitrogen-based systems: a bimolecular case and some qualitative aspects. AB - The experimentally observed reversible hydrogen activation reaction by trans-2,6 dimethyl-2,6-diphenylpiperidine and B(C(6)F(5))(3) is modeled at RI-SCS-MP2/aug cc-pVTZ//B3LYP/6-31G* level. A Morokuma analysis is performed for the transition state and for the product to study the energy contributions. The role of solvent effects, and substitution of ligands is discussed. Some more general points are made on 'frustration' (meaning the prevention of B-N bond formation by bulky ligands) and 'Coulomb pays for Heitler-London' (meaning that the attraction between the product counterions is comparable with the homolytic splitting energy of H(2)). Both ideas refer to hypothetical deviations from the experimental route. A simple NH(3) + H(2) + BH(3)<--> (NH)(BH) model reaction is used to estimate the frustration energy and to locate the ionic/covalent crossings of this 'bimolecular' model. The ionic structures are stabilised further in oligomers (and crystals), a phenomenon termed as 'collective Madelung ionisation' (CMI) and illustrated by a [(NH)(BH)](4) tetramer. PMID- 20024455 TI - First-principles investigations of Ti-substituted hydroxyapatite electronic structure. AB - The electronic structure of Ti-substituted hydroxyapatite is investigated using density functional theory within a periodic slab model. Two sorption mechanisms have been considered: i.e., Ti(4+) and Ti(OH)(2)(2+) as the likely species to exchange with Ca(2+). Ti(4+) has a small ionic radius compared to Ca(2+) and can dope into both distinct sites, showing no site preference; however, when two H were removed from the OH channel to obtain charge compensation, preferential site II substitution appears, accompanied with a large O shift forming a strong Ti-O bond. The species Ti(OH)(2)(2+) displays a strong site preference: substitution by Ti(OH)(2)(2+) on the hydroxyl channel (site II) is exothermic and favored strongly over the Ca column (site I). Ti(OH)(2)(2+) substitution for Ca(2+) induces a large geometry relaxation and distortion, especially within the OH channel and Ca(2+) column, with a considerable shift of Ti compared to the Ca sites in pure HA. These results are consistent with the experimental observation that material synthesis with high Ti doping (atomic ratio > 0.1) shows irregular particles formation with reduced crystallinity. The calculated cell shape and volume relaxations indicate that the volume and cell parameters both expand in all the substituted HA models. The site preference and volume expansion differences found are attributed to the metal ion shift caused in meeting the requirement of strong Ti-O coordination in site I and site II polyhedra. PMID- 20024456 TI - Enhancing the interactions between neutral molecular tweezers and anions. AB - Several structural modifications to the original molecular tweezers of the Klarner's group were made with the aim of improving its binding capacity towards anions. The proposed modifications raise the molecular electrostatic potential inside the cavity and provide more conformational flexibility. The complexes of these new molecules with the halide anions Cl(-), Br(-), I(-) were optimized at the MPW1B95/6-31+G* level of theory. The molecular interactions were analyzed by single point density fitted local second-order Moller-Plesset perturbation theory (DF-LMP2) and DF-LMP2 spin-component-scaled MP2 (SCS-MP2), calculations were performed with the cc-pVTZ basis set. In view of the large magnitude of the interaction energies computed and the stability of the complexes in different solvents, this kind of molecule is a good candidate as molecular host for anion recognition. PMID- 20024457 TI - Molecular dynamics simulations of atomically flat and nanoporous electrodes with a molten salt electrolyte. AB - The electric double layer (EDL) structure and capacitance have been studied for atomically flat and nanoporous conductive electrodes with a molten LiCl electrolyte using an electroactive interface molecular dynamics simulation methodology. For the atomically flat electrodes the electrolyte was observed to form a multilayer structure near the electrode described by exponentially decaying sinusoidal oscillations in ion and charge densities perpendicular to the electrode/electrolyte interface. The differential EDL capacitance vs. electrode potential was found to exhibit "U-shaped" behavior while the EDL capacitance exhibited complex dependence on electrode potential including regions of negative capacitance near zero electrode potential. Increased capacitance and an enhanced degree of electrode-electrolyte interface structure were observed with decreasing temperature. For nanoporous electrodes with both slit and cylindrical pore geometries, the electrolyte was observed to form highly structured alternating charged layers within the electrode nanopores. A maximum in the normalized (per unit electrode area) EDL capacitance was found for pore widths that accommodate several charged layers inside the pores. The observed dependence of capacitance on pore size appears to be a compromise between increasing structure/charge imbalance and decreasing ion density with decreasing pore width/diameter. PMID- 20024458 TI - The influence of alkaline earth ions on the structural organization of acetone probed by the noncoincidence effect of the nu(C=O) band: experimental and quantum chemical results. AB - We have investigated the Raman noncoincidence effect (NCE = nu(aniso)-nu(iso), where nu(aniso) and nu(iso) are the anisotropic and the isotropic Raman frequencies) of the nu(C=O) band of acetone arising from the interactions of this solvent with the metal ions in acetone electrolytic solutions of alkaline earth metal (Mg, Ca, Sr, Ba) perchlorates. Assisted by the results of ab initio molecular orbital (MO) calculations carried out at the Hartree-Fock (HF) level with the 6-31+G(2df,p) and LanL2DZ basis sets, we have been able to attribute the anisotropic and isotropic components of this band to the formation of acetone metal ion clusters, (acetone)(n)M(2+), and to interpret its high and negative NCE, opposed to the positive NCE of the bulk liquid, as the consequence of the large separation between the higher frequency of the in-phase mode (active in the Raman isotropic spectrum) and the lower (average) frequency of the n- 1 out-of phase modes (predominantly active in the Raman anisotropic spectrum). The negative sign of the NCE is compatible with the transition dipole coupling (TDC) mechanism. The comparison between the observed NCE for each electrolytic solution at the concentrations used in this study and those calculated for the different solvation numbers n of each (acetone)(n)M(2+)cluster gives a clear indication of the highest stability of the hexa-coordinated cluster for the Mg(2+) ion, but leaving uncertain (n = 6 or 8) this conclusion for the acetone clusters of the remaining M(2+) ions. We have interpreted the observed and calculated decrease of the magnitude of NCE with the ion size through the ion polarizing power in the light of the ion effective charge and its distance (M(2+)...O=C) from the C=O oscillators. PMID- 20024459 TI - Carotenoids can act as antioxidants by oxidizing the superoxide radical anion. AB - The electron transfer (ET) reaction between carotenoids and the superoxide radical anion is found to be not only a viable process but also a very unique one. The nature of the O(2) (-) inverts the direction of the transfer, with respect to ET involving other ROS: the O(2) (-) becomes the electron donor and carotenoids (CAR) the electron acceptor. Therefore the "antioxidant" activity of CAR when reacting with O(2) (-) lies in their capacity to prevent the formation of oxidant ROS. This peculiar charge transfer is energetically feasible in non polar environments but not in polar media. In addition the relative reactivity of CAR towards O(2) (-) is drastically different from their reactivity to other ROS. Asthaxanthin (ASTA) is predicted to be a better O(2) (-) quencher than LYC and the other CAR. The CAR + O(2) (-) reactions were found to be diffusion controlled. The agreement with available experimental data supports the density functional theory results from the present work. PMID- 20024460 TI - Electron spin relaxation at low field. AB - The low field ESR lineshape and the electron spin-lattice relaxation correlation function are calculated using the stochastic Liouville theory for an effective electron spin quantum number S = 1. When an axially symmetric permanent zero field splitting provides the dominant relaxation mechanism, and when it is much larger than the rotational diffusion constant, it is shown that both electron spin correlation functions S(0)S(t) (n = 0,1) are characterized by the same relaxation time tau(S) = (4D(R))(-1). This confirms the conjectures made by Schaefle and Sharp, J. Chem. Phys., 2004, 121, 5287 and by Fries and Belorizky, J. Chem. Phys., 2005, 123, 124510, based on numerical results using a different formalism. The stochastic Liouville approach also gives the paramagnetically enhanced nuclear spin relaxation time constants, T(1) and T(2), and the ESR lineshape function I(omega). In particular, the L-band (B(0) = 0.035 T) ESR spectrum of a low symmetry Ni(ii)-complex with a cylindrical ZFS tensor is shown to be detectable at sufficiently slowly reorientation of the complex. The analysis shows that the L-band spectrum becomes similar to the zero-field spectrum with a electron spin relaxation time tau(S) = (4D(R))(-1). PMID- 20024461 TI - Polarizable continuum model associated with the self-consistent-reaction field for molecular adsorbates at the interface. AB - In this work, a new procedure has been developed in order to realize the self consistent-reaction field computation for interfacial molecules. Based on the extension of the dielectric polarizable continuum model, the quantum-continuum calculations for interfacial molecules have been carried out. This work presents an investigation into how the molecular structure influences the adsorbate solvent interaction and consequently alters the orientation angle at the air/water interface. Taking both electrostatic and non-electrostatic energies into account, we investigate the orientation behavior of three interfacial molecules, 2,6-dimethyl-4-hydroxy-benzonitrile, 3,5-dimethyl-4-hydroxy benzonitrile and p-cyanophenol, at the air/water interface. The results show that the hydrophilic hydroxyl groups in 2,6-dimethyl-4-hydroxy-benzonitrile and in p cyanophenol point from the air to the water side, but the hydroxyl group in 3,5 dimethyl-4-hydroxy-benzonitrile takes the opposite direction. Our detailed analysis reveals that the opposite orientation of 3,5-dimethyl-4-hydroxy benzonitrile results mainly from the cavitation energy. The different orientations of the hydrophilic hydroxyl group indicate the competition of electrostatic and cavitation energies. The theoretical prediction gives a satisfied explanation of the most recent sum frequency generation measurement for these molecules at the interface. PMID- 20024462 TI - Thermodynamics of nano- and macrocrystalline anatase using cell voltage measurements. AB - In view of increasing scientific and technological interest in nanomaterials, it is important to examine whether or, more exactly, to what extent the thermodynamic parameters change with size. Electrochemical e.m.f. measurements which provide a direct and elegant access to these thermodynamic data have been used in this study to investigate the excess contributions of anatase due to nano size. The e.m.f. measurements are carried out (250-450 degrees C) on different particle sizes (1.2 microm-5 nm) using the cell: Au, O(2), Na(2)Ti(6)O(13), TiO(2) (anatase) |Na-beta'' alumina |TiO(2) (rutile), Na(2)Ti(6)O(13), O(2), Au. The e.m.f. observed is closely related to the difference of the Gibbs energies of formation (Delta(f)G degrees) of the titania crystals on both sides. Such cell voltage measurements with various sizes of anatase (1200, 100, 15, and 5 nm) as working electrodes enable us to calculate the excess enthalpy and entropy due to surface contributions and to provide refined data for the macroscopic anatase. No electrochemical Ostwald ripening or chemical Ostwald ripening was observed in the case of anatase nanoparticles up to 500 degrees C. PMID- 20024463 TI - Valence electron energy-loss spectroscopy of silicon negative electrodes for lithium batteries. AB - All compounds present in the lithium-silicon binary phase diagram were synthesized and analyzed by electron energy-loss spectroscopy. In order to limit beam damage, and to develop a fast and local method of characterizing silicon negative electrodes, the valence energy-loss spectrum region was investigated, in particular the very intense plasmon peak in these alloys. Experimental spectra are in strong agreement with theoretical ones obtained from density functional theory. These results constitute a database for Li(x)Si alloys' plasmon energies. The method is applied to the study of the first discharge of a silicon electrode, thus identifying a Li(2.9+/-0.3)Si phase in equilibrium with Si on the voltage plateau. A nucleation process of this phase in the pristine Si is revealed, as well as a possible over-lithiation beyond the end of discharge Li(15)Si(4) crystalline phase. PMID- 20024464 TI - Spectroscopic characterization of Fe-doped synthetic chrysotile by EPR, DRS and magnetic susceptibility measurements. AB - Fe-doped synthetic geomimetic chrysotile nanocrystals represent a reference standard to investigate the health hazard associated with asbestos fibers and constitute interesting inorganic nanotubes for specific technological applications in light harvesting systems, optoelectronics and photonics. As the fiber toxicity is catalyzed by iron ions in specific crystallographic sites and the mechanical behaviour of synthetic chrysotile nanotubes is strongly affected by the iron doping extent, the characterization of Fe substitution to Mg and/or Si sites in the chrysotile structure appears highly important. By EPR, DRS spectroscopic analyses and magnetic investigations, Mg and/or Si ion replacement by Fe(3+) in a synthetic geomimetic chrysotile structure has been investigated. The results highlight that, as a function of the Fe doping extent and of the Fe doping process, iron can replace both Mg and Si sites. The contemporary iron substitution into the octahedral and tetrahedral sheets is associated with the presence of both of isolated Fe(3+) centres in high-spin 3d(5) configuration (S = 5/2, (6)A(1)((6)S)) in O(h) and T(d) symmetry and of intra-lattice clustered species. Increasing the Fe doping extent increases the concentration of aggregated species, while magnetic susceptibility confirms a paramagnetic anisotropy. The results allow to define the opportunity of using or not metallic Fe during the synthesis to obtain doped chrysotile nanocrystals with tailored morphological and structural properties suitable as a reference to study asbestos toxicity and apt to prepare new inorganic nanotubes and quantum wires for innovative technological applications. PMID- 20024465 TI - Dealloying to nanoporous Au/Pt alloys and their structure sensitive electrocatalytic properties. AB - A simple and general dealloying method is employed to fabricate nanoporous Au/Pt alloys with pre-determined alloy compositions. Structural characterization by electron microscopes demonstrates that selective etching of Cu from Au/Pt/Cu alloy precursors results in the formation of three-dimensional bicontinuous porous network structures with uniform pores and ligaments less than 10 nm. X-Ray photoelectron spectroscopy and X-ray diffraction demonstrate that nanoporous Au/Pt alloys have a single-phase cubic structure with relatively uniform compositions across the samples. These high surface area alloy nanostructures show much enhanced specific activity and distinct surface reactivity toward the electrooxidation of some small organic molecules, such as methanol and formic acid, as the Au content varies within the structure, thus holding great potential for use in clean energy and environmental applications. PMID- 20024466 TI - Mass spectrometry and beam deflection studies of tin-lead nanoalloy clusters. AB - Photo-ionization mass spectrometry and electric beam deflection experiments were used to study isolated Sn(M)Pb(N) clusters (7 85-90%, intraluminal thrombus, ICA tortuosity, gross surface ulceration of plaque and excessive calcification. Type III aortic arch and arch calcification also precluded CAS. Standard techniques were used for both procedures with a protection device routinely used for CAS. Most CEAs were performed under general anaesthesia, with selective intraluminal shunting. One hundred and eighty-six patients were selected for CAS; nine (48% ) were converted to CEA for technical reasons. The operative risk profile was similar, but significantly more in the CAS group were hypertensive. Almost half (49% ) in the CAS group were asymptomatic vs 26% in the CEA group. All asymptomatics had 70+ % stenosis on Duplex Doppler. Results were reported within one month of the procedure. The stroke rate was 2.3% for CAS vs 1.9% for CEA (p > 0.05). Stroke and death plus one M1 was 4.5% after CAS vs 3.4% after CEA (p > 0.05). Disabling stroke occurred in 1.1% of CAS patients vs 0.4% of CEA patients. These results are satisfactory and compare favourably with other similar series. PMID- 20024473 TI - Evaluation of the reliability of the EuroSCORE risk analysis prediction in high risk older patients undergoing CABG. AB - BACKGROUND: The aim of this prospective study was to evaluate the reliability of EuroSCORE risk-analysis predictions on early mortality in high-risk older patients who underwent heart surgery. METHODS: From January 2008 to February 2009, a total of 128 consecutive high-risk older patients who underwent open heart surgery were included. Patients who required emergency surgery, had pulmonary hypertension, a recent myocardial infarction, underwent combined heart surgery procedures or had renal disease were included. The patients had a mean age of 72 +/- 9 years (range 64-91, 53.1% male) and were evaluated for surgery. RESULTS: Coronary artery bypass graft (CABG) surgery was performed on 112 patients and valve surgery on 16. Eight patients (6.25% ) died in hospital. The observed mortality rate was lower than the expected mortality obtained using EuroSCORE (6.25% vs 11.2 +/- 7.2%, respectively, p < 0.021). CONCLUSION: There was no correlation between the preoperative logarithmic score of expected mortality and the observed mortality rate in these older high-risk patients who underwent open-heart surgery. PMID- 20024474 TI - Oscillometric blood pressure reference values of African full-term neonates in their first days postpartum. AB - BACKGROUND: Knowing the normative blood pressure (BP) in a newborn baby is important in order to identify abnormal BP readings. This study was done to determine normative BP values of Nigerian newborns, using the 8100 Dinamap monitor. METHODS: Consecutive full-term neonates delivered in a tertiary centre in Nigeria were recruited for the study. The babies' systolic (SBP), diastolic (DBP) and mean arterial (MAP) blood pressures were measured within the first four days after birth. RESULTS: A total of 473 babies were recruited for the study. The mean SBP, DBP and MAP readings on day 1 were 66.8 +/- 7.7, 38.5 +/- 6.3 and 47.9 +/- 6.3 mmHg, respectively. The day 1 SBP of babies > 4 kg were significantly higher than those who weighed < 2.5 and 2.5-4 kg (p = 0.01, p = 0.05), respectively. CONCLUSION: This study provided current normative SBP, DBP and MAP values for Nigerian neonates. The BP readings compared with their Caucasian counterparts. PMID- 20024475 TI - Pattern of heart failure in Abuja, Nigeria: an echocardiographic study. AB - AIM: Despite heart failure having been identified in subjects in sub-Saharan Africa over the last 60 years, there is still a dearth of data, especially echocardiographic data on heart failure. We therefore set out to analyse the clinical and echocardiographic features of all consecutive subjects presenting with heart failure in a tertiary institution in Nigeria. METHODS: Three hundred and forty subjects with heart failure, according to the guidelines of the European Society of Cardiology, were studied. Each patient had two-dimensional guided transthoracic echocardiography. RESULTS: The mean age of the patients was 50.60 +/- 15.29 years, and 50.9% of the study population were males while 49.1% were females. The commonest cause of heart failure identified was hypertension in 61.5% of the patients; 75.5% had systolic heart failure, whereas 23.5% had heart failure with preserved ejection fraction. CONCLUSIONS: Untreated hypertension has been identified as the leading cause of heart failure in Abuja, Nigeria, which is similar to that in many other parts of sub-Saharan Africa. Coronary artery disease is a rare cause of heart failure in this population group. PMID- 20024476 TI - Percutaneous closure of a secundum atrial septal defect after surgical pericardectomy. AB - A case of successful closure of a percutaneous atrial septal defect following surgical pericardectomy is described. Clinical and haemodynamic aspects of atrial septal defects associated with constrictive pericarditis are also discussed. PMID- 20024477 TI - An unusual case of an obtuse marginal branch supplying the left ventricular apex. AB - In most patients, the left anterior descending coronary artery (LAD) feeds the anterior aspect of the interventricular septum and a large segment of the anterior wall of the left ventricle. It then passes around the left ventricular apex to supply that area as well. However, in some cases, the LAD is shorter and terminates either at or before the left ventricular apex. The latter in these cases is instead supplied by a long posterior descending coronary artery (PDA), originating from the right coronary artery (RCA) at the crux, then passing along the entire length of the posterior interventricular groove, and either terminating at the apex or extending beyond it to pass upward in the anterior interventricular groove. In this case report, we describe a 50-year-old man with an apical perfusion defect on stress myocardial perfusion imaging in whom diagnostic coronary angiography revealed that the left ventricular apex was supplied by a large, obtuse marginal branch of the left circumflex coronary artery (LCX) instead of the LAD. PMID- 20024478 TI - Late presentation of aorto-right ventricular fistula and associated aortic regurgitation following penetrating chest trauma. AB - We present a case in which a 39-year-old man was assaulted in 2002 and sustained penetrating chest trauma. He was treated conservatively, and presented six years later with exertional dyspnoea and a continuous murmur. Echocardiography detected an aorta-right ventricular fistula and an important aortic regurgitation. The aorta-right ventricular fistula was repaired and the aortic valve was replaced with a mechanical prosthesis. Traumatic aorto-right ventricular fistulas and aortic regurgitation are often masked by the primary injury and may take months or years to become clinically evident. PMID- 20024479 TI - An unintentional left ventricular stimulation. AB - A 31-year-old man had undergone pacemaker implantation for complete atrio ventricular block six years before. Permanent ventricular stimulation was seen on the ECG. The QRS complex was negative in leads I, VL and V2 to V5. The ventricular lead tip was found to be screwed into a branch of the great cardiac vein. Unintentional coronary sinus fixed-screw lead implantation has not to our knowledge been reported before. PMID- 20024480 TI - Localised dissection in the convexity of the aortic arch. AB - We present images of a chest CT scan of an 89-year-old man, showing an area of localised dissection in the convexity of the aortic arch. The ascending and descending aorta were not involved in the disease. The patient suffered a stroke, with severe neurological impairment. Differential diagnosis was undertaken. PMID- 20024481 TI - Prediction of potential protein post-translational modifications of the thioredoxin-1 molecule. AB - Thioredoxin-1 is a 12-kDa protein with anti-oxidative, antiapoptotic and pro proliferative functions, and it offers protection against cardiac hypertrophy. It has been found that the physiological regulation of thioredoxin-1 requires post translational modifications and this may be an important step in the pathophysiology of cardiomyopathy. In this study, the potential protein post translational modifications in thioredoxin-1 were determined with a standard bioinformatics technique, FindMod. No potential protein posttranslational modifications were found, which may indicate that they are not an important step in the pathophysiology of cardiomyopathy. PMID- 20024482 TI - Servier Programme for Registrars in Cardiology (SPARC). PMID- 20024489 TI - Protein Z and protein Z-dependent protease inhibitor and renal tubules. PMID- 20024490 TI - Protective effect of ischaemic preconditioning on ischaemia/reperfusion-induced microvascular obstruction determined by on-line measurements of coronary pressure and blood flow in pigs. AB - We investigated the protective effect of ischaemic preconditioning (IP) on the maintenance of coronary patency using on-line measurements of coronary pressures and blood flow in a closed-chest reperfused acute myocardial infarction (MI) model in pigs. Catheter-based 90-min occlusion followed by 60-min reperfusion of the left anterior descending coronary artery (LAD) was performed in anesthetised pigs (MI group). IP was applied (IP group) through two cycles of 5-min occlusion and 5-min reperfusion of the LAD before MI induction. Coronary patency was determined by measurements of coronary wedge pressure, collateral fractional flow reserve (FFRcoll), collateral pressure index (CPI) and absolute coronary blood flow (CBF). Inducible and constitutive nitric oxide synthase (iNOS/cNOS) activities and expressions were determined in the myocardium. Plasma levels of myeloperoxidase (MPO, index of activated leukocytes) and mean platelet volume (MPV, index of activated platelets) were measured. IP resulted in significantly lower levels of MPO (0.52 +/- 0.19 vs. 1.05 +/- 0.24 U/l, p<0.001) and MPV (9.1 +/- 0.6 vs. 9.6 +/- 1.0 fl, p=0.04), higher FFRcoll (0.17 +/- 0.05 vs. 0.04 +/- 0.05, p<0.001), CPI (0.13 +/- 0.05 vs. 0.02 +/- 0.05, p<0.001) and CBF (70.7 +/- 4.2 vs. 50.8 +/- 4.8 m/min, p<0.001) post-reperfusion as compared with the MI group. IP resulted in significantly higher cNOS activity and eNOS expression. Significant negative correlation was found between MPO and measures of coronary patency (FFRcoll, CPI and CBF) and cNOS activity. Moreover, cNOS activity correlated significantly with FFRcoll, CPI and CBF. In conclusion, IP attenuates the release of MPO and platelet activation, thereby contributing to the maintenance of vessel patency at microvascular level after reperfusion of the infarct-related artery. PMID- 20024491 TI - Intra-patient variability of thromboelastographic parameters following in vivo and ex vivo administration of recombinant activated factor VII in haemophilia patients. A multi-centre, randomised trial. AB - Thromboelastography methods have been used to predict or monitor treatment of haemophilia patients with recombinant activated factor VII (rFVIIa). However, neither of the two thromboelastographic methods (ROTEM and TEG) has as yet been validated. This multi-centre, randomised trial compared both methods in terms of intra- and inter- patient variability following in vivo and ex vivo rFVIIa administration to haemophilia A and B patients with and without inhibitors. Patients ((3)16 years old) received the same intravenous rFVIIa dose (45, 90 or 180 microg/kg) twice, 1-12 weeks apart. Blood samples were collected pre-dose and 15, 60, 120 and 240 minutes post-dose for ROTEM and TEG analysis. Pre-dose samples were also spiked ex vivo with rFVIIa (0.6, 1.2 or 2.4 microg/ml), to correspond to the three in vivo doses. Twenty-six haemophilia A and four haemophilia B patients were enrolled. A significant treatment effect was observed with in vivo rFVIIa (p<0.05) with more pronounced effects in inhibitor (n=14) versus non-inhibitor (n=16) patients. There was a strong positive correlation between ROTEM and TEG parameters. Intra- and inter-patient variation was large for all thromboelastography parameters at all time points and rFVIIa doses. Intra patient variation was generally lower for non-inhibitor than inhibitor patients, and lower following ex vivo spiking versus in vivo rFVIIa administration. In conclusion, there was a clear effect of rFVIIa on all thromboelastography parameters, but the large intra- and inter-patient variability following in vivo rFVIIa administration renders the use of our method unsuitable for dose-response prediction for haemophilia patients in the clinical setting. PMID- 20024492 TI - What is the appropriate approach to prevention of thromboembolism in heart failure? AB - Many studies suggest a higher incidence of thromboembolic syndromes such as stroke, peripheral arterial thrombosis and pulmonary embolism in patients with heart failure (HF), particularly those with left ventricular systolic dysfunction. As a result, some clinicians have chosen to treat patients with HF with anticoagulants as primary prevention against thromboembolic events. However, this practice is not well-supported by scientific data. Retrospective analyses of large HF trials have yielded contradictory results and randomised trials designed to specifically address this question have been under-populated and under powered. As a result, there is no general consensus among professional societies in either recommending or advising against anticoagulants in HF. We hope that ongoing clinical trials, WARCEF in particular, will yield results that will guide clinicians in deciding for or against routine use of anticoagulants in HF. PMID- 20024493 TI - Activation of platelets in young infants during cardiopulmonary bypass. PMID- 20024494 TI - Loss of high-molecular-weight von Willebrand factor multimers mainly affects platelet aggregation in patients with aortic stenosis. AB - Severe aortic stenosis is associated with a haemostatic abnormality that resembles acquired von Willebrand syndrome type 2. It is assumed that high shear conditions render large von Willebrand factor (VWF) multimers accessible to cleavage by ADAMTS-13. However, whether loss of these large multimers affects platelet function by impairing adhesion, aggregate formation, or both has not been evaluated in clinical studies. We prospectively enrolled 47 patients with severe aortic stenosis, and studied them prior to aortic valve surgery and at a median of six months after valve replacement. We investigated levels of large VWF multimers, platelet function under high shear conditions, and residual response to suboptimal concentrations of ADP to express P-selectin. As expected, there was a significant reduction of VWF large multimers before surgery that resolved thereafter in most patients (p<0.0001). The closure time of the ADP cartridge of the PFA-100 was also corrected in most patients after the operation (p<0.0001). We used the cone and plate(let) analyser Impact-R to differentiate between adhesion and aggregation. Both adhesion (p=0.03) and ADP-inducible platelet aggregation (p=0.002) improved considerably after valve replacement. Consequently, ADP-inducible expression of P-selectin was higher after valve replacement (p=0.001). We conclude that reduced levels of large VWF multimers associated with aortic stenosis lead to impairment of both adhesion and, especially, ADP-inducible platelet aggregation. PMID- 20024496 TI - Frequency, demographics and risk (according to tumour type or site) of cancer associated thrombosis among patients seen at outpatient DVT clinics. AB - Venous thromboembolism (VTE) is a clinically important complication for both hospitalised and ambulatory cancer patients. In the current study, the frequency, demographics and risk (according to tumour site) of VTE were examined among patients seen at outpatient DVT (deep-vein thrombosis) clinics. Of 10,015 VTE cases, 1,361 were diagnosed with cancer, for an overall rate of cancer-associated VTE of 13.6% in this outpatient population. Patients with cancer-associated VTE were significantly older than cancer-free VTE cases (66.4 +/- 12.7 vs. 58.8 +/- 18.5 years; p<0.0001). The frequency of cancer-associated VTE peaked earlier among females than males, occurring in the sixth (137/639, 21.4% vs. 98/851, 11.3%; p<0.001) and seventh decades (213/980, 21.7% vs. 197/1096, 18%; p=0.036). VTE was described most frequently in common cancers - breast, prostate, colorectal and lung (56.1% of cases). The risk of VTE varied widely across 17 cancer types. Calculating odds ratios (OR) to assess the effect size of cancer type on VTE risk, the highest odds were observed for patients with pancreatic cancer (OR 9.65, 95% confidence interval [CI] (5.51-16.91). Tumours of the head and neck had higher odds than previously reported (OR 8.24, 95% CI 5.06-13.42). Reduced risk estimates were observed for skin cancers (melanoma and non-melanoma: OR 0.89, 95% CI 0.42-1.87; OR 0.74, 95% CI, 0.32-1.69, respectively). We conclude that outpatients have a similar rate of cancer-associated VTE as VTE patient populations previously reported, that cancer-associated VTE occurs in an older age group and earlier in females and that outpatients exhibit distinct tumour site-specific risk from that described among hospitalised cancer patients. PMID- 20024495 TI - Impaired thrombin generation and fibrin clot formation in patients with dilutional coagulopathy during major surgery. AB - Patients subjected to haemodilution during surgery are at increased risk of bleeding. We hypothesised that, in the acquired dilutional coagulopathy, insufficient haemostasis is due to either insufficient thrombin generation or insufficient fibrin clot formation. In tissue factor-activated plasmas from patients with coagulation deficiency, we measured time curves of thrombin generation and fibrin clot formation (thromboelastography). Investigated were in study A: 10 patients treated with vitamin K antagonist and five healthy subjects; in study B: 30 patients undergoing cardiopulmonary bypass (CPB) surgery and infused with on average 2,000 ml crystalloids and colloids (no major bleeding); in study C: 58 patients undergoing major general surgery, and transfused with >5,000 ml crystalloids, colloids and red cell concentrates, who experienced major bleeding and were post-transfused with fresh frozen plasma. The treatment with vitamin K antagonist led to a progressive reduction in thrombin generation but not fibrin clot formation. In CPB patients, plasma factor levels post-surgery were 53-60% of normal. This was accompanied by moderate reduction in both haemostatic processes. In plasmas from patients undergoing major surgery, factor levels were 38-41% of normal, and these levels increased after plasma transfusion. Taking preset thresholds for normal thrombin generation and fibrin clot formation, at least one of these processes was low in 88-93% of the patients with (persistent) bleeding, but only in 40-53% of the patients without bleeding. In conclusion, the ability of thrombin generation and fibrin clot formation is independently reduced in acquired dilutional coagulopathy, while minimal levels of both are required for adequate haemostasis. PMID- 20024497 TI - Proteolytic resistance conferred to fibrinogen by von Willebrand factor. AB - The formation of platelet-rich thrombi under high shear rates requires both fibrinogen and von Willebrand factor (VWF) as molecular adhesives between platelets. We attempted to describe the role of VWF as a potential substrate and modulator of the fibrinolytic system using binding assays, as well as kinetic measurements on the cleavage of fibrin(ogen) and a synthetic plasmin substrate (Spectrozyme-PL). The similar dissociation constants for the binding of plasminogen, plasmin, and active site-blocked plasmin onto immobilised VWF suggest that the primary binding site in plasmin(ogen) is not the active site. The progressive loss of clottability and generation of degradation products during fibrinogen digestion with plasmin were delayed in the presence of VWF at physiological concentrations, while VWF cleavage was not detectable. Determination of kinetic parameters for fibrinogen degradation by plasmin, miniplasmin and microplasmin showed that VWF did not modify the Km, whereas kcat values decreased with increasing VWF concentrations following the kinetic model of non-competitive inhibition. Inhibitory constants calculated for VWF were in the range of its physiological plasma concentration (5.4 mg/ml, 5.7 mg/ml and 10.0 mg/ml for plasmin, miniplasmin and microplasmin, respectively) and their values suggested a modulating role of the kringle 5 domain in the interaction between VWF and (mini)plasmin. VWF had no effect on the amidolytic activity of plasmin on Spectrozyme-PL, or on fibrin dissolution by (mini)plasmin. Our data suggest that VWF, while a poor plasmin substrate relative to fibrinogen, protects fibrinogen against degradation by plasmin preserving its clottability in plasma and its adhesive role in platelet-rich thrombi. PMID- 20024498 TI - Toll-like receptor 2 stimulation of platelets is mediated by purinergic P2X1 dependent Ca2+ mobilisation, cyclooxygenase and purinergic P2Y1 and P2Y12 receptor activation. AB - Toll-like receptor 2 (TLR2), which recognise and respond to conserved microbial pathogen-associated molecular patterns, is expressed on the platelet surface. Furthermore, it has recently been shown that the TLR2/1 agonist Pam3CSK4 stimulates platelet activation. The aim of the present study was to clarify important signalling events in Pam3CSK4-induced platelet aggregation and secretion. Platelet interaction with Pam3CSK4 and the TLR2/6 agonist MALP-2 was studied by analysing aggregation, ATP-secretion, [Ca2+]i mobilisation and thromboxane B2 (TxB2) production. The results show that Pam3CSK4 but not MALP-2 induces [Ca2+]i increase, TxB2 production, dense granule secretion and platelet aggregation. Preincubation of platelets with MALP-2 inhibited the Pam3CSK4 induced responses. The ATP-secretion and aggregation in Pam3CSK4-stimulated platelets was impeded by the purinergic P2X1 inhibitor MRS 2159, the purinergic P2Y1 and P2Y12 antagonists MRS 2179 and cangrelor, the phospholipase C inhibitor U73122, the calcium chelator BAPT-AM and aspirin. The calcium mobilisation was lowered by MRS 2159, aspirin and U73122 whereas the TxB2 production was antagonised by MRS 2159, aspirin and BAPT-AM. When investigating the involvement of the myeloid differentiation factor-88 (MyD88) -dependent pathway, we found that platelets express MyD88 and interleukin 1 receptor-associated kinase (IRAK 1), which are proteins important in TLR signalling. However, Pam3CSK4 did not stimulate a rapid (within 10 minutes) phosphorylation of IRAK-1 in platelets. In conclusion, the results show that Pam3CSK4-induced platelet aggregation and secretion depends on a P2X1-mediated Ca2+ mobilisation, production of TxA2 and ADP receptor activation. The findings in this study further support a role for platelets in sensing bacterial components. PMID- 20024499 TI - Anticoagulation for valvular heart disease in community-based practice. AB - Little is known about patients who receive oral anticoagulation for valvular heart disease (VHD) in community-based practice. It was this study's objective to describe the characteristics, management, and outcomes of patients anticoagulated for VHD, compared to patients anticoagulated for atrial fibrillation (AF). We used a nationally-representative cohort of community-based anticoagulation care in the United States. Data collected included indications for therapy, demographics, selected comorbid conditions, international normalised ratio (INR) target ranges, INR control, and clinical outcomes. We identified 1,057 patients anticoagulated for VHD (15.6% of the overall cohort) and 3,396 patients anticoagulated for AF (50.2%). INR variability was similar between the two groups (0.64 vs. 0.69, p = 0.80). Among patients with aortic VHD, for whom a standard (2 3) target INR range is recommended, 461 (84%) had a high target range (2.5-3.5), while 95 (16%) had a standard target range. VHD patients had a higher rate of major haemorrhage compared to AF patients (3.57 vs. 1.78 events per 100 patient years, incidence rate ratio 2.02, 95% CI 1.33 - 3.06). The rate of stroke/systemic embolus was similar between groups (0.67 vs. 0.97 events per 100 patient-years, incidence rate ratio 0.71, 95% CI 0.32 - 1.57). In our community based study, approximately 15.6% of patients receiving warfarin were anticoagulated for VHD. VHD patients achieved similar anticoagulation control to patients with AF, as measured by INR variability. Nevertheless, the rate of major haemorrhage was elevated among VHD patients compared to AF patients; this finding requires further investigation. PMID- 20024500 TI - Sulfated, low-molecular-weight lignins are potent inhibitorsof plasmin, in addition to thrombin and factor Xa: Novel opportunity for controlling complex pathologies. AB - Recently we prepared sulfated, low-molecular-weight lignins (LMWLs) to mimic the biological activities of heparin and heparan sulfate. Chemo-enzymatically prepared sulfated LMWLs represent a library of diverse non-sugar, aromatic molecules with structures radically different from the heparins, and have been found to potently inhibit thrombin and factor Xa. To assess their effect on the fibrinolytic system, we studied the interaction of LMWLs with human plasmin. Enzyme inhibition studies indicate that the three sulfated LMWLs studied inhibit plasmin with IC50 values in the range of 0.24 and 1.3 mM, which are marginally affected in the presence of antithrombin. Similarly, plasmin degradation of polymeric fibrin is also inhibited by sulfated LMWLs. Michaelis-Menten kinetic studies indicate that maximal velocity of hydrolysis of chromogenic substrates decreases nearly 70% in the presence of LMWLs, while the effect on Michaelis constant is dependent on the nature of the substrate. Competitive binding studies indicate that the sulfated LMWLs compete with full-length heparin. Comparison with thrombin-heparin crystal structure identifies an anionic region on plasmin as a plausible sulfated LMWL binding site. Overall, the chemo-enzymatic origin coupled with coagulation and fibrinolysis inhibition properties of sulfated LMWLs present novel opportunities for designing new pharmaceutical agents that regulate complex pathologies in which both systems are known to play important roles such as disseminated intravascular coagulation. PMID- 20024501 TI - Economic evaluation of dabigatran etexilate for the prevention of venous thromboembolism in patients aged over 75 years or with moderate renal impairment undergoing total knee or hip replacement. AB - Oral dabigatran etexilate is indicated for the prevention of venous thromboembolism (VTE) in patients undergoing total knee replacement or total hip replacement. We investigated the cost-effectiveness of the 150 mg once daily (od) dose recommended for patients aged over 75 or with moderate renal impairment, from a United Kingdom National Health Service perspective. Dabigatran etexilate was compared with subcutaneous enoxaparin 40 mg od, using a decision model. Risks for VTE and bleeding were derived from subgroup analyses of the phase III trials. Dabigatran etexilate was less costly than enoxaparin; cost savings varied from pound62 to pound274 (base-case analyses) and were primarily due to differences in administration costs. Results were robust across a range of sensitivity analyses. Dabigatran etexilate 150 mg od is cost saving compared with enoxaparin 40 mg od in patients aged over 75years and in patients with moderate renal impairment, with comparable efficacy and safety. PMID- 20024503 TI - Thrombophilia and repeated in vitro fertilisation and embryo transfer failure: an open issue. PMID- 20024502 TI - Contribution of exosite occupancy by heparin to the regulation of coagulation proteases by antithrombin. AB - Heparin promotes the antithrombin (AT) inactivation of factors IXa (fIXa) and Xa (fXa) through a conformational activation of the serpin and also by a template mechanism in the presence of physiological levels of Ca2+. Recently, it was reported that heparin induces conformational changes in the active-sites of fIXa and fXa, raising the possibility that heparin also modulates the reactivity of these proteases with AT by this mechanism. To test this possibility, we prepared an AT mutant in which four critical heparin-binding residues of the serpin (Arg 45, Arg-46, Lys-114, and Arg-129) were replaced with non-basic residues. This mutant lost its affinity for heparin, but retained its normal reactivity with coagulation proteases. Thus, the high-affinity AT-binding pentasaccharide fragment of heparin had no cofactor effect on the reactivity of the AT mutant with coagulation proteases. Full-length heparin-concentration dependence of the AT inhibition of fIXa and fXa revealed that in contrast to a greater than 4-5 orders of magnitude accelerating effect for heparin on the AT inhibition of fIXa and fXa, heparin exhibits a negligible cofactor effect (<2-fold) on the mutant AT inhibition of these proteases. The same results were obtained for the mutant AT inhibition of thrombin and factor VIIa, however, heparin accelerated the mutant AT inhibition of factor XIa ~10-fold. We conclude that, with the exception of factor XIa, heparin-mediated conformational modulation of the active-sites of coagulation proteases makes a minor contribution to the regulation of these proteases by AT. PMID- 20024504 TI - Endogenous thrombin potential, prothrombin fragment 1+2 and D-dimers during pregnancy. PMID- 20024505 TI - The TRITON versus PLATO trials: differences beyond platelet inhibition. AB - Clopidogrel monopoly as an exclusive oral antiplatelet agent used in combination with aspirin or as a monotherapy for treatment or/and prevention of occlusive thrombotic vascular events has been recently challenged. Based on the indirect comparison of TRITON and PLATO trial data, ticagrelor is clearly superior to prasugrel in a population of patients with acute coronary syndrome (ACS) because of absolute mortality reduction, realistic second myocardial infarction (MI) prevention, growing over time vascular outcome benefit, fewer haemorrhagic fatalities, potentially less coronary artery bypass graft (CABG)- related bleeding events, and lack of cancer risks. Despite an unfavourable immediate safety profile, ticagrelor has a lot of room to compensate for agitation, dyspnea, and ventricular pauses, if used in appropriate patients. It will be naive and wrong to assume that ticagrelor will completely substitute clopidogrel, especially considering higher discontinuation rates after ticagrelor, generic competition, and other health economics issues. However, unless the regulatory authorities discover some unexpected serious flaws with PLATO, the ticagrelor will substantially change the present landscape of oral antiplatelet therapy, especially in high-risk patients, diabetics, and those with repeated vascular events including stent thrombosis. In contrast, a too exclusive trial design, a lack of persistent vascular benefit despite issues with event adjudication, growing-over-time bleeding complications, an issue with cancer, and finally an increase in mortality risk among unstable angina and non ST-elevated myocardial infarction will likely prevent a broad prasugrel implementation, unless more reassuring evidence becomes available. PMID- 20024506 TI - Heyde's syndrome: From controversy to mainstream. PMID- 20024508 TI - Horseshoe kidney in an 80 year old with chronic kidney disease. AB - This image illustrates a horseshoe kidney from an elderly man that was being evaluated for chronic kidney disease associated with diabetes. Horseshoe kidney is the most common renal fusion abnormality, with an incidence of 1/400 worldwide and a male-to-female ratio of 2:1 and is not a cause of impaired kidney function. One theory of the abnormal fusion of the kidneys is that the metanephric blastemas close together before renal capsule maturation, resulting in fusion. The normal embryonic ascent of the fused kidney is disrupted by the inferior mesenteric artery, leaving the adult horseshoe kidney in a lower position than normal. About 90% of the time, the lower poles are fused and the excretory renal units and ureters are maintained separately. PMID- 20024507 TI - Human solCD39 inhibits injury-induced development of neointimal hyperplasia. AB - Blood platelets provide the initial response to vascular endothelial injury, becoming activated as they adhere to the injured site. Activated platelets recruit leukocytes, and initiate proliferation and migration of vascular smooth muscle cells (SMC) within the injured vessel wall, leading to development of neointimal hyperplasia. Endothelial CD39/NTPDase1 and recombinant solCD39 rapidly metabolise nucleotides, including stimulatory ADP released from activated platelets, thereby suppressing additional platelet reactivity. Using a murine model of vascular endothelial injury, we investigated whether circulating human solCD39 could reduce platelet activation and accumulation, thus abating leukocyte infiltration and neointimal formation following vascular damage. Intraperitoneally-administered solCD39 ADPase activity in plasma peaked 1 hour (h) post-injection, with an elimination half-life of 43 h. Accordingly, mice were administered solCD39 or saline 1 h prior to vessel injury, then either sacrificed 24 h post-injury or treated with solCD39 or saline (three times weekly) for an additional 18 days. Twenty-four hours post-injury, solCD39-treated mice displayed a reduction in platelet activation and recruitment, P-selectin expression, and leukocyte accumulation in the arterial lumen. Furthermore, repeated administration of solCD39 modulated the late stage of vascular injury by suppressing leukocyte deposition, macrophage infiltration and smooth muscle cell (SMC) proliferation/migration, resulting in abrogation of neointimal thickening. In contrast, injured femoral arteries of saline-injected mice exhibited massive platelet thrombus formation, marked P-selectin expression, and leukocyte infiltration. Pronounced neointimal growth with macrophage and SMC accretion was also observed (intimal-to-medial area ratio 1.56 +/- 0.34 at 19 days). Thus, systemic administration of solCD39 profoundly affects injury-induced cellular responses, minimising platelet deposition and leukocyte recruitment, and suppressing neointimal hyperplasia. PMID- 20024509 TI - A case of immunotactoid glomerulopathy with rapid progression to end-stage renal disease. AB - Immunotactoid glomerulopathy (IGN) is a rare immunoglobulin deposition disease. It is often mistaken for cryoglobulinemia or amyloidosis due to the similarities on biopsy findings. The disease progresses to end-stage renal disease (ESRD) within 7 months to 10 years. This is the first case reported of a patient with a diagnosis of IGN who developed acute kidney injury (AKI) and ESRD within 1 week of initial presentation. PMID- 20024510 TI - Calciphylaxis mimicking penile gangrene: a case report. AB - Calciphylaxis is a rare, but life-threatening, disease, mostly seen in patients with renal failure, especially those undergoing dialysis. It is characterized by violaceous tender areas of cutaneous plaques, necrosis, and eschar formation, mostly involving toes and fingers, but rarely the penis. Peripheral pulses are mostly preserved. The parathyroid hormone (PTH) level is elevated, along with raised calcium phosphorus product. There is radiological evidence of blood vessel and soft tissue calcification. Predisposing factors are obesity and diabetes. It is rarely encountered by a urologist and closely resembles penile gangrene. PMID- 20024511 TI - Molecular markers in upper urothelial carcinoma associated to Balkan endemic nephropathy. Aristolochic acid as the major risk factor of the worldwide disease. AB - The role of aristolochic acid in the etiology of Balkan endemic nephropathy (BEN) and associated upper urothelial carcinoma (UUC) was recently confirmed. The aim of this study was to determine the marker(s) specific for BEN-associated UUC. A total of 82 patients with UUC (38 from the BEN region and 44 control tumors) were included in the study. The Ki-67 index in BEN tumors correlated with the grade and multifocality (p < 0.05), but in regression analysis, only the grade of BEN tumor. The p53 index was significantly higher in BEN than in control tumors (p < 0.05), as well as the alteration of p53 (p < 0.05). BEN low-stage tumors, tumors without limphovascular invasion (LVI), and tumors of the renal pelvis had a higher p53 index than the control tumors (p < 0.05, 0.01, 0.05, respectively). The Ki-67 index was higher in control tumors with high-stage and solid growth than in BEN UUC (p < 0.050, 0.005). The Ki-67 correlated with the grade, growth, stage, LVI, and multifocality of UUC on the best way, but not with the group. In regression analysis, only multifocality of UUC had predictive influence on Ki-67 activity (p < 0.001). P53 correlated with the grade, growth, and group (p < 0.05). This investigation identifies the p53 pathway as the specific cell cycle marker involved in BEN-associated UUC. PMID- 20024512 TI - Induced abortion: risk factors for adolescent female students, a Brazilian study. AB - The purpose of this study was to analyze risk factors for abortion among female teenagers from 12 to 19 years of age in the city of Maceio, Brazil. This is a cross-sectional study, conducted in ten schools. The sample was calculated by considering the number of admissions for postabortion curettage, obtained from the Information System of Hospitalization. Data were obtained through a semi structured questionnaire divided into three basic blocks of data: sociodemographic, sexual life, and pregnancy/abortion. To analyze the data, the logistic regression model was used. The Forward Method was chosen to set the final model that minimizes the number of variables and maximizes the accuracy of the model. The significant analysis between the dichotomous variables provided eight significant variables. Two of them are protective for abortion: the ages 12 14 years and talking with parents about sex. After the logistic regression, the receipt of support for abortion was the most significant variable of all. The adolescent with an active sexual life, a previous pregnancy, who is married, and has received support for an abortion has a 99.74% probability for an abortion. The results of this study, demonstrating the importance of the group in adolescence, and the statistical significance of having a partner to support and approve the pregnancy appears as a preventive factor for abortion. It shows the importance of support and companionship for adolescent women. PMID- 20024513 TI - Influence of poly-(L-lactic acid) nanofiber functionalization on maximum load, Young's modulus, and strain of nanofiber scaffolds before and after cultivation of osteoblasts: an in vitro study. AB - The aim of this study was to characterize the influence of functionalization of synthetic poly-(L-lactic acid) (PLLA) nanofibers on mechanical properties such as maximum load, elongation, and Young's modulus. Furthermore, the impact of osteoblast growth on the various nanofiber scaffolds stability was determined. Nanofiber matrices composed of PLLA, PLLA-collagen, or BMP-2-incorporated PLLA were produced from different solvents by electrospinning. Standardized test samples of each nanofiber scaffold were subjected to failure protocol before or after incubation in the presence of osteoblasts over a period of 22 days under osteoinductive conditions. PLLA nanofibers electrospun from hexafluoroisopropanol (HFIP) showed a higher strain and tended to have increased maximum loads and Young s modulus compared to PLLA fibers spun from dichloromethane. In addition, they had a higher resistance during incubation in the presence of cells. Functionalization by incorporation of growth factors increased Young's modulus, independent of the solvent used. However, the incorporation of growth factors using the HFIP system resulted in a loss of strain. Similar results were observed when PLLA was blended with different ratios of collagen. Summarizing the results, this study indicates that different functionalization strategies influence the mechanical stability of PLLA nanofibers. Therefore, an optimization of nanofibers should not only account for the optimization of biological effects on cells, but also has to consider the stability of the scaffold. PMID- 20024514 TI - Melanocortin peptide therapy for the treatment of arthritic pathologies. AB - Arthritic pathologies are a major cause of morbidity within the western world, with rheumatoid arthritis affecting approximately 1% of adults. This review highlights the therapeutic potential of naturally occurring hormones and their peptides, in both arthritic models of disease and patients. The arthritides represent a group of closely related pathologies in which cytokines, joint destruction, and leukocytes play a causal role. Here we discuss the role of naturally occurring pro-opiomelanocortin (POMC)-derived melanocortin peptides (e.g., alpha melanocyte stimulating hormone [alpha-MSH]) and synthetic derivatives in these diseases. Melanocortins exhibit their biological efficacy by modulating proinflammatory cytokines and subsequent leukocyte extravasation. Their biological effects are mediated via seven transmembrane G-protein-coupled receptors, of which five have been cloned, identified, and termed MC1 to MC5. Adrenocorticotrophic hormone represents the parent molecule of the melanocortins; the first 13 amino acids of which (termed alpha-MSH) have been shown to be the most pharmacologically active region of the parent hormone. The melanocortin peptides have been shown to display potent anti-inflammatory effects in both animal models of disease and patients. The potential anti-inflammatory role for endogenous peptides in arthritic pathologies is in its infancy. The ability to inhibit leukocyte migration, release of cytokines, and induction of anti inflammatory proteins appears to play an important role in affording protection in arthritic injury, and thus may lead to potential therapeutic targets. PMID- 20024515 TI - Mirror-like mechanisms and music. AB - The neural processes underlying sensory-motor integration have always attracted strong interest. The classic view is that action and perception are two extremes of mental operations. In the past 2 decades, though, a large number of discoveries have indeed refuted such an interpretation in favor of a more integrated view. Specifically, the discovery of mirror neurons in monkey premotor cortex is a rather strong demonstration that sensory and motor processes share the same neural substrates. In fact, these cells show complex sensory-motor properties, such that observed, heard, or executed goal-directed actions could equally activate these neurons. On the other hand, the neuroscience of music has similarly emerged as an active and productive field of research. In fact, music related behaviors are a useful model of action-perception mechanisms and how they develop through training. More recently, these two lines of research have begun to intersect into a novel branch of research. As a consequence, it has been proposed recently that mirror-like mechanisms might be at the basis of human music perception-production abilities. The scope of the present short review is to set the scientific background for mirror-like mechanisms in music by examining recent published data. PMID- 20024516 TI - Visualizing vertebrate embryos with episcopic 3D imaging techniques. AB - The creation of highly detailed, three-dimensional (3D) computer models is essential in order to understand the evolution and development of vertebrate embryos, and the pathogenesis of hereditary diseases. A still-increasing number of methods allow for generating digital volume data sets as the basis of virtual 3D computer models. This work aims to provide a brief overview about modern volume data-generation techniques, focusing on episcopic 3D imaging methods. The technical principles, advantages, and problems of episcopic 3D imaging are described. The strengths and weaknesses in its ability to visualize embryo anatomy and labeled gene product patterns, specifically, are discussed. PMID- 20024517 TI - Trastuzumab resistance: role for Notch signaling. AB - Epidermal growth factor receptor-2 (ErbB-2/HER2) is a potent breast oncogene that has been shown to be amplified in 20% of breast cancers. Overexpression of ErbB-2 predicts for aggressive tumor behavior, resistance to some cytotoxic and antihormonal therapies, and poor overall survival. Trastuzumab, the humanized, monoclonal antibody directed against ErbB-2 has shown tremendous efficacy and improved overall survival for women when combined with a taxane-based chemotherapy. However, resistance to trastuzumab remains a major concern, most notably in women with metastatic breast cancer. Numerous mechanisms that include overexpression of alternate receptor tyrosine kinases and/or loss of critical tumor suppressors have been proposed in the last several years to elucidate trastuzumab resistance. Here we review the many possible mechanisms of action that could contribute to resistance, and novel therapies to prevent or reverse the resistant phenotype. Moreover, we provide a critical role for Notch signaling cross-talk with overlapping or new signaling networks in trastuzumab-resistant breast. PMID- 20024519 TI - Lewy body variant of Alzheimer's disease: selective neocortical loss of t-SNARE proteins and loss of MAP2 and alpha-synuclein in medial temporal lobe. AB - Lewy bodies (LBs) appear in the brains of nondemented individuals and also occur in a range of neurodegenerative disorders, such as dementia with Lewy bodies (DLB) and Parkinson's disease. A number of people with a definite diagnosis of Alzheimer's disease (AD) also exhibit these intraneuronal inclusions in allo- and/or neocortical areas. The latter, referred to as Lewy body variant of AD (LBV), bears a clinical resemblance to AD in terms of age at onset, duration of illness, cognitive impairment, and illness severity. Since the presence of LBs is accompanied by neuronal cytoskeleton changes, it is possible that the latter may influence neuronal connectivity via alterations to the synaptic network. To address this, we examined the expression of synaptic proteins (synaptophysin, syntaxin, SNAP-25, and alpha-synuclein) and two cytoskeletal proteins (tau and MAP2) in the brain tissue of subjects enrolled in a population-based autopsy study (n = 47). They were divided into groups with no memory problems (control group, n = 15), LBV (n = 5), AD devoid of LBs (n = 17), cerebrovascular dementia (n = 3), and mixed dementia (n = 7). The LBV and AD groups had a similar degree of cognitive impairment and neuropathological staging in terms of Braak staging and CERAD score. In comparison with the control group and the dementia groups without LBs, the LBV group had significantly lower levels of syntaxin and SNAP-25 (23%) in the neocortex, and depletion of MAP2 (64%), SNAP-25 (34%), and alpha synuclein (44%) proteins in the medial temporal lobes. These findings suggest that the t-SNARE complex deficit present in LBV may be associated with the presence of LB-related pathology and may explain the more profound cholinergic loss seen in these patients. PMID- 20024520 TI - Molecular biology of rotavirus entry and replication. AB - Rotavirus is a nonenveloped, double-stranded, RNA virus belonging to the Reoviridae family and is the major etiological agent of viral gastroenteritis in young children and young animals. Remarkable progress in the understanding of the rotavirus cycle has been made in the last 10 years. The knowledge of viral replication thus far acquired is based on structural studies, the expression and coexpression of individual viral proteins, silencing of individual genes by siRNAs, and the effects that these manipulations have on the physiology of the infected cell. The functions of the individual rotavirus proteins have been largely dissected; however, the interactions between them and with cell proteins, and the molecular mechanisms of virus replication, are just beginning to be understood. These advancements represent the basis for the development of effective vaccination and rational therapeutic strategies to combat rotavirus infection and diarrhea syndromes. In this paper, we review and try to integrate the new knowledge about rotavirus entry, replication, and assembly, and pose some of the questions that remain to be solved. PMID- 20024521 TI - Cancer risk among Japanese chromium platers, 1976-2003. AB - OBJECTIVE: The aim of our prospective cohort study was to assess cancer mortality risks among chromium platers. METHODS: The cohort comprised 1193 male platers (626 with exposure to chromium, the remainder with no exposure) with a follow-up period of 27 years (1 October 1976 to 31 December 2003). Mortality risk was assessed by the standardized mortality rate (SMR) with reference to the national population. RESULTS: Lung cancer mortality was elevated only in the chromium plater subgroup, with borderline statistical significance [SMR=1.46, observations (Obs)=28, 95% confidence interval (95% CI) 0.98-2.04]. The chromium plater subgroup also showed elevated mortality risks for brain tumor (SMR=9.14, Obs=3, 95% CI 1.81-22.09) and malignant lymphoma (SMR=2.84, Obs=6, 95% CI 1.05-5.51). Risks were particularly elevated for lung cancer (SMR=1.59, Obs=23, 95% CI 1.01 2.38) and malignant lymphoma (SMR=3.80, Obs=6, 95% CI 1.39-8.29) among those with initial chromium exposure prior to 1970. CONCLUSIONS: In Japan, occupational exposure to chromium through work as a chromium plater is a risk factor for lung cancer, especially for platers working prior to 1970. Occupational chromium exposure may also increase the risk of brain tumor and malignant lymphoma. PMID- 20024518 TI - Altered nuclear functions in progeroid syndromes: a paradigm for aging research. AB - Syndromes of accelerated aging could provide an entry point for identifying and dissecting the cellular pathways that are involved in the development of age related pathologies in the general population. However, their usefulness for aging research has been controversial, as it has been argued that these diseases do not faithfully reflect the process of natural aging. Here we review recent findings on the molecular basis of two progeroid diseases, Werner syndrome (WS) and Hutchinson-Gilford progeria syndrome (HGPS), and highlight functional connections to cellular processes that may contribute to normal aging. PMID- 20024522 TI - Occupation and the risk of hearing impairment--results from the Nord-Trondelag study on hearing loss. AB - OBJECTIVES: We studied the effect of occupation on hearing and if it remained after adjustment for noise exposure, education, income, and other risk factors. METHODS: Audiometry and a questionnaire concerning exposure was administered to a general adult population sample in Norway (N=49,948). Information on occupation, education, and income was obtained from population census registers. RESULTS: Occupation had marked effects on hearing loss. Occupation explained 2-3% of the variance in hearing loss among men > or =45 years in addition to the hearing loss due to age (10-19%). Occupation explained < or =1% of hearing loss among women of all ages and young men. Controlling for self-reported occupational noise exposure reduced the occupational effect by 20-40% in men > or =45 years. Controlling for leisure-time noise, ear infections, and head injuries did not change the effect of occupation, which was slightly reduced after controlling for education and income. The most elevated hearing thresholds in men were observed among: wood workers; miners; linemen and cable jointers; construction carpenters and workers; seamen; and workshop mechanics. CONCLUSIONS: There was a moderate association between occupation and hearing loss. Unbiased estimates of occupational hearing loss may help identify high-risk occupations, for which interventions are needed, and identify individuals with hearing loss. PMID- 20024523 TI - [Phlegmon of the palm of the hand as initial manifestation of the Lemierre syndrome]. AB - Lemierre's syndrome is a rare disease in young otherwise healthy people showing septic embolism in the lungs and peripheral vessels. We report the case of a 19 year-old male patient who presented initially with a phlegmon of the right palm and beginning septic shock. During the clinical course a subcutaneous abscess of the left shoulder, multiple lesions of the lungs and a pericardial abscess were identified and Lemierre's syndrome was diagnosed. In this case, positron emission tomography (PET) was revealed to be an appropriate instrument to determine the extent of the disease in a one step procedure. PMID- 20024524 TI - [Chronic osteitis of the lower extremities. An interdisciplinary treatment concept]. AB - INTRODUCTION: The purpose of this investigation was the retrospective analysis of patients with delayed infections, chronic posttraumatic osteitis of the lower extremities and free-flap coverage after radical debridement of bone and soft tissue. METHODS: From the time period 1994-2003 a total of 22 patients including 4 females and 18 males were investigated. In 16 patients treatment was carried out on the lower leg and in 6 patients the foot was treated with subsequent free flap coverage. In 14 cases the latissimus dorsi muscle was used, in 5 cases the gracilis muscle, in 2 cases parascapula flaps were used and in 1 case the serratus anterior muscle. The average age of the patients was 43 years (range 17 63 years) and grouping was according to the HOST classification. Functional outcome was evaluated by a standardized questionnaire (Funktionsfragebogen Hannover FFbH-OA 2,0), quality of life and social reintegration by non standardized questionnaires. RESULTS: In the cases investigated the following results could be achieved: full leg activity 55%, leg pain while walking 73%, special footwear 68%, normal gait 55%, positive quality of life and social reintegration 55%, port activities 36% and reemployment 45%. CONCLUSION: According to the results of this study the quality of life of patients with chronic osteitis of the lower leg is in general satisfying. In order to improve quality management and cost reduction in public health an interdisciplinary treatment concept of plastic and orthopedic surgeons should be established for complex fracture management as this is the most effective tool in treating chronic osteitis. PMID- 20024525 TI - [Biomechanical investigation of fixed-angle plate osteosynthesis of the proximal humerus]. AB - INTRODUCTION: Proximal humeral fractures are common in the elderly population and are often associated with osteoporosis. Fixation of unstable proximal humeral fractures is problematic due to loss of fixation in osteoporotic bone. Fixed angle devices are intended to provide superior mechanical stability due to the principle of an internal fixator. The NCB(R)-PH (non-contact-bridging proximal humerus) plate is a new fixed-angle device that locks the screws to the plate by secondary insertion of a locking cap. The aim of this study was to investigate if and to what extent NCB-PH plates applied in the locked mode provide higher mechanical stability in a proximal humerus fracture model. METHODS: For this investigation 16 (8 pairs) fresh frozen cadaveric humeri were used. An AO/ASIF 11 B 1 fracture of the proximal humerus was created in each bone and fixed with the NCB-PH plate. In a paired setting one bone was fixed with the plate in the locked mode and the other with the plate in the non-locked mode. The bones were then subjected to 100 cycles of axial loading and interfragmentary motion was measured. Bones surviving this test were subjected to load-to-failure testing and the force at which failure occurred was recorded. RESULTS: Bones fixed with the plate in the locked mode showed a statistically significant lower (51%) interfragmentary rotation compared to bones fixed with the plate in the non locked mode. There was also a tendency for the bones fixed with the plate in the locked mode to fail first under higher forces (16%) during load-to-failure testing. CONCLUSION: The NCB-PH plate provides superior interfragmentary stability when used in the locked mode in a human cadaveric proximal humerus fracture model. Therefore, we recommend that all screws should be inserted in the locked mode. The results suggest that the NCB-PH plate in the locked mode provides higher primary postoperative stability thus permitting early functional treatment of the patient. PMID- 20024526 TI - [Guideline compliance in the treatment of schizophrenic patients. Introduction of a computer-assisted treatment pathway]. AB - BACKGROUND: The goal of S3 Guidelines for the Treatment of Schizophrenia was to improve the care of patients with schizophrenic psychoses. However, the publication of guidelines alone does not ensure their consistent implementation. The use of treatment pathways represents one possible approach to help implement the complex treatment recommendations contained in the S3 Guidelines. The first computer-assisted treatment pathway for patients with schizophrenic psychoses was successfully incorporated into the everyday routine of psychiatric hospitals. The aim of the present study was to systematically analyse the impact of this measure on guideline compliance. MATERIALS AND METHODS: Based on the S3 Guidelines for the Treatment of Schizophrenia developed by the German Association of Psychiatry, Psychotherapy and Neurology (Deutsche Gesellschaft fur Psychiatrie, Psychotherapie und Nervenheilkunde; DGPPN), diagnostic and treatment procedures were defined by a multiprofessional working group with members from five different hospitals and subsequently incorporated into an existing hospital information system. In one of the five hospitals, the impact of this measure was analysed in a pilot study in a systematic manner. In the year 2007, approximately 100 patients in each of two wards in the hospital received in a parallel group design either standard care or care based on a computer-assisted treatment pathway. Based on their place of residence, patients were assigned to the two units consecutively. Both groups were analysed to determine the extent to which the care they received conformed to treatment guidelines. Data available from the years 2004 and 2005 served as a historical comparison to the present results. RESULTS: The differences in guideline compliance between the two wards were heterogeneous and, in certain respects, counterintuitive. As expected, the treatment pathway group showed an increased number of laboratory tests, more frequent drug screening at hospital admission and more appropriate dosing of neuroleptics. However, the rate of participation in psychoeducational interventions was disappointing. A conspicuous finding was the negative relationship between initial disease severity and compliance with guidelines on psychopharmacological treatment. In contrast, the historical comparison revealed that guideline compliance had increased slightly in both the treatment pathway and standard treatment groups. CONCLUSION: Developing computer-assisted treatment pathways based on S3 Guidelines and incorporating them into existing hospital information systems is feasible and well accepted by users. The initial effects on guideline compliance are mostly positive, but not strongly so. Moreover, there was a reduction in duration of hospital stay. Disease-related factors such as disease severity appear to compromise guideline compliance. PMID- 20024527 TI - [Magnetoencephalography in psychiatry]. AB - Neuropsychiatric disorders usually come with only sublime structural changes. Functional imaging can point at specific disturbances in information processing in neural networks. Besides imaging of receptor and metabolic functions with PET and fMRI, electromagnetic methods such as electroencephalography (EEG) and magnetoencephalography (MEG) offer the possibility for imaging of dynamic dysfunctions. As compared to EEG, MEG has a shorter history and is less common despite offering considerable advantages in temporospatial resolution and sensitivity to detect impaired signal processing and network functioning which renders it particularly interesting for psychiatric applications. Disturbed processing in the auditory and visual domain emerging in schizophrenic, affective and anxiety disorders can be detected with high sensitivity. Moreover, the neuromagnetic baseline activity allows conclusions to be drawn regarding neural network functions. Due to its high sensitivity to single deficits in information processing and to pharmacological effects, MEG will achieve clinical significance in specific areas. PMID- 20024528 TI - Cardioinhibitory properties of potato glycoalkaloids in beetles. AB - The semi-isolated heart bioassay was used to evaluate the effect of glycoalkaloids extracted from potato leaves on the heart contractile activity of three beetle species Zophobas atratus, Tenebrio molitor and Leptinotarsa decemlineata. The dose-response curves indicated species specific action of tested substances. Application of glycoalkaloids on the continuously perfused Z. atratus heart inhibited progressively frequency contractions; higher concentrations exerted short and reversible cardiac arrests. In the rest two beetle species tested glycoalkaloids caused no cardiotropic effect. In vivo bioassay with 1 day old Z. atratus pupae showed that the extract induces a negative inotropic effect on the heart. PMID- 20024529 TI - Bone health and deterioration in quality of life among participants from the Hertfordshire cohort study. AB - We utilised the Hertfordshire cohort study to examine relationships between bone density at baseline and SF-36 status 4 years later. We found deterioration in the mental health domain over follow-up in osteoporotic men (but not women) compared with other groups (relative rate ratio=5.78, 95% confidence interval (CI) 1.78 19.2). INTRODUCTION: Osteoporosis is associated with decreased quality of life, although it has been difficult to evaluate the confounding effects of fracture and co-morbidity. Having previously shown that male osteoporotics have poorer health than counterparts with normal bone mineral density, even after adjustment for co-morbidity and prior fracture, we assessed quality of life in both groups 4 years apart. METHODS: Four hundred and ninety-eight men and 468 women completed questionnaires detailing lifestyle factors, co-morbidities and quality of life (SF-36) before undergoing bone density measurements at the lumbar spine and total femur. At follow-up 4 years later, 322 men and 320 women were reassessed. RESULTS: Multinomial logistic regression confirmed deterioration in mental health over follow-up in osteoporotic men compared with other groups (relative rate ratio=5.78, 95% CI 1.78-19.2). These patterns were not apparent among women. CONCLUSIONS: Men with lower bone density at baseline had poorer quality of life some 4 years later, even after adjustment for co-morbidity and fracture. This may reflect secondary osteoporosis in men (due to alcohol or hypogonadism). PMID- 20024531 TI - Chemical characterization of engineered nanoparticles. PMID- 20024530 TI - Spore-to-spore agar culture of the myxomycete Physarum globuliferum. AB - The ontogeny of the myxomycete Physarum globuliferum was observed on corn meal agar and hanging drop cultures without adding sterile oat flakes, bacteria or other microorganisms. Its complete life cycle including spore germination, myxamoebae, swarm cells, plasmodial development, and maturity of fructifications was demonstrated. Details of spore-to-spore development are described and illustrated. PMID- 20024532 TI - Determination of the natural abundance delta15N of nortropane alkaloids by gas chromatography-isotope ratio mass spectrometry of their ethylcarbamate esters. AB - An important route for the detoxification of tropane alkaloids involves N demethylation to the nor-compounds followed by further degradation. In order to study the mechanisms of the pertinent reactions, a suitable means to determine the isotope ratios of the substrates and products is required. However, the polarity and functionality of the nortropane compounds makes their analysis as free bases difficult. A method is described which allows both the quantification of nortropane alkaloids and the determination of their natural abundance delta(15)N values. The protocol exploits the derivatisation of the alkaloids by reaction with ethyl chloroformate in aqueous medium and the quantitative extraction of the ensuing ethylcarbamate esters. The improved chromatographic properties of these derivatives gives ample separation of the isomeric nortropine and norpseudotropine for measurement of their delta(15)N (per thousand) values by isotope ratio mass spectrometry interfaced to gas chromatography. Adequate separation could not be achieved with the underivatised compounds. Repeatability and precision are sufficient to allow differences in the delta(15)N values (Deltadelta(15)N) > 0.8 per thousand to be measured, with a standard deviation routinely approximately 0.3 per thousand. The methodology has been tested by determining the changes in the delta(15)N values of nortropine and norpseudotropine during degradation by cell suspension cultures of a Pseudomonas strain expressing a specific capacity for tropine catabolism. The precision and reproducibility are shown sufficient to allow the evolution of the delta(15)N values to be followed during the fermentation. PMID- 20024533 TI - Quantitative LC-ESI-MS/MS metabolic profiling method for fatty acids and lipophilic metabolites in fermentation broths from beta-lactam antibiotics production. AB - In the present paper, we report on the development of a straightforward reversed phase liquid chromatography-electrospray ionization-tandem mass spectrometry method for the determination of the most abundant fatty acids; alpha-tocopherol and cephalosporin P1 in fermentation broths. Using this method, fatty acids could be successfully determined in extracts of fermentation broths from penicillin and cephalosporin production without prior derivatization. Matrix effects were investigated in detail, and various kinds of calibrations (i.e., by use of neat standard solutions as well as by matrix-matched calibration employing standard addition each with and without internal standards) were comparatively assessed. The optimized and validated method was employed for the analysis of extracts of fermentation broths and nutrition media. PMID- 20024534 TI - Phthalates determination in pharmaceutical formulae used in parenteral nutrition by LC-ES-MS: importance in public health. AB - A method for determining a group of phthalate esters in pharmaceutical formulae used in parenteral nutrition samples (with and without vitamins) has been developed. The phthalic acid esters (PAEs) studied were dimethyl phthalate, diethyl phthalate, butyl benzyl phthalate, dibutyl phthalate, di-(2-ethylhexyl) phthalate, and dioctyl phthalate. This group of phthalates was determined by high performance liquid chromatography (HPLC)-electrospray ionization-mass spectrometry, working in positive ion mode. The phthalates analyzed were extracted from the sample using hexane and sodium hydroxide. The hexane was then evaporated, and the compounds were redissolved in acetonitrile. The compounds were separated by HPLC working in gradient mode with acetonitrile-ultrapure water starting from 5% to 75% acetonitrile in 5 min, followed by isocratic elution for 27 min. Standard calibration curves were linear for all the analytes over the concentration range 10-250 microg L(-1). The method was precise (with RSD from 3.3% to 12.9%) and sensitive. The proposed analytical method has been applied to the analysis of these compounds in different pharmaceutical formulae (with different compositions) for parenteral nutrition samples in order to check the presence of phthalates and determine their concentration. PMID- 20024535 TI - The effect of long-term danazol prophylaxis on liver function in hereditary angioedema-a longitudinal study. AB - BACKGROUND: Danazol is a drug most widely used for the prophylaxis of hereditary angioedema resulting from the deficiency of the C1-inhibitor. Potential hepatotoxic or liver tumor-inducing side effects of long-term danazol prophylaxis have been investigated during the follow-up of hereditary angioedema patients. METHODS: Characteristic parameters of liver function (including bilirubin, GOT, GPT, gammaGT, total protein, ALP, LDH), as well as findings of viral serology screens and abdominal ultrasonography-determined during years 0 and 5 of follow up of patient groups taking/not taking danazol-have been reviewed and analyzed comparatively. RESULTS: From a population of 126 hereditary angioedema patients, 46 subjects taking danazol and another 46 not taking danazol fulfilled the inclusion criteria. Longitudinal follow-up did not reveal any clinically relevant difference between the liver function parameters determined in years 0 and 5 in the two groups. Abdominal ultrasound did not detect neoplastic or other potentially treatment-related alterations of the liver parenchyma. There were no discontinuations of treatment during the study. CONCLUSIONS: Our results clearly suggest that, administered at the lowest effective dose, danazol does not induce liver injury in hereditary angioedema patients. PMID- 20024536 TI - Drug dependence associated with triptans and ergot derivatives: a case/non-case study. AB - INTRODUCTION: The aim of this case/non-case study was to assess and compare the risk of drug dependence associated with different migraine-specific drugs, i.e., ergot derivatives and triptans, using the French pharmacovigilance database. METHODS: Reports on drug side effects recorded in this database between January 1985 and June 2007 were analyzed, and triptans (almotriptan, eletriptan, naratriptan, sumatriptan, and zolmitriptan) as well as ergot derivatives used in acute migraine were examined. For all reports, cases were defined as those reports corresponding to "drug abuse," "physical or mental drug dependence," and "pharmacodependence," whereas "non-cases" were defined as all the remaining SED reports. The method's reliability was assessed by calculating the risk associated with a negative (amoxicillin) and a positive (benzodiazepines) control. The risk of dependence associated with each drug and control was evaluated by calculating the odds ratio (OR) with a confidence interval of 95%. RESULTS: Among the 309,178 reports recorded in the database, drug dependence accounted for 0.8% (2,489) of the reports, with 10.9% (449) involving a triptan, and 9.33% (332) an ergot derivative. The risk of dependence was similar for triptans and ergot derivatives and did not differ from that of benzodiazepines. In the triptan group, the risk (odds ratio [95% CI]) ranged from 10.3 [4.8-22.3] for sumatriptan to 21.5 for eletriptan [10.1-45.6], while in the ergot derivative group, it ranged from 12 [8 17.9] for ergotamine to 20.6 [8-53] for dihydroergotamine. CONCLUSIONS: These findings confirm the hypothesis that triptans and ergot derivatives are associated with an increased risk of drug dependence. PMID- 20024537 TI - The phylogenetic distribution of non-CTCF insulator proteins is limited to insects and reveals that BEAF-32 is Drosophila lineage specific. AB - Chromatin insulators are DNA sequences found in eukaryotes that may organize genomes into chromatin domains by blocking enhancer-promoter interactions and preventing heterochromatin spreading. Considering that insulators play important roles in organizing higher order chromatin structure and modulating gene expression, very little is known about their phylogenetic distribution. To date, six insulators and their associated proteins have been characterized, including Su(Hw), Zw5, CTCF, GAF, Mod(mdg4), and BEAF-32. However, all insulator proteins, with the exception of CTCF, which has also been identified in vertebrates and worms, have been exclusively described in Drosophila melanogaster. In this work, we have performed database searches utilizing each D. melanogaster insulator protein as a query to find orthologs in other organisms, revealing that except for CTCF all known insulator proteins are restricted to insects. In particular, the boundary element-associated factor of 32 kDa (BEAF-32), which binds to thousands of sites throughout the genome, was only found in the Drosophila lineage. Accordingly, we also found a significant bias of BEAF-32 binding sites in relation to transcription start sites (TSSs) in D. melanogaster but not in Anopheles gambiae, Apis mellifera, or Tribolium castaneum. These data suggest that DNA binding proteins such as BEAF-32 may have a dramatic impact in the genome of single evolutionary lineages. A more thorough evaluation of the phylogenetic distribution of insulator proteins will allow for a better understanding of whether the mechanism by which these proteins exert their function is conserved across phyla and their impact in genome evolution. PMID- 20024538 TI - Evolutionary rates in Veronica L. (Plantaginaceae): disentangling the influence of life history and breeding system. AB - The evolutionary rate at which DNA sequences evolve is known to differ between different groups of organisms. However, the reasons for these different rates are seldom known. Among plants, the generation-time hypothesis, which states that organisms that reproduce faster also have more DNA substitutions per time, has gained most popularity. We evaluate the generation-time hypothesis using 131 DNA sequences from the plastid trnLF region and the nuclear ribosomal ITS region of the genus Veronica (Plantaginaceae). We also examine the alternative hypothesis that a higher substitution rate is correlated with selfing breeding system. Selfing is associated with annual life history in many organisms and may thus often be the underlying reason for observed correlations of annual life history with other characters. We provide evidence that annual life history is more likely to be the responsible factor for higher substitution rates in Veronica than a selfing breeding system. Nevertheless, the way in which annual life history may influence substitution rate in detail remains unknown, and some possibilities are discussed. PMID- 20024539 TI - The origin of conserved protein domains and amino acid repeats via adaptive competition for control over amino acid residues. AB - Some proteins, such as homeodomain transcription factors, contain highly conserved regions of sequence. It has recently been suggested that multiple functional domains overlap in the homeodomain, together explaining this high conservation. However, the question remains why so many functional domains cluster together in one relatively small and constrained region of the protein. Here we have modeled an evolutionary mechanism that can produce this kind of clustering: conserved functional domains are displaced from the parts of the molecule that are undergoing adaptive evolution because novel functions generally out-compete conserved functions for control over the identity of amino acid residues. We call this model COAA, for Competition Over Amino Acids. We also studied the evolution of amino acid repeats (a.k.a. homopeptides), which are especially prevalent in transcription factors. Repeats that are encoded by non homogenous mixtures of synonymous codons cannot be explained by replication slippage alone. Our model provides two explanations for their origin, maintenance, and over-representation in highly conserved proteins. We demonstrate that either competition between multiple functional domains for space within a sequence, or reuse of a sequence for many functions over time, can cause the evolution of amino acid repeats. Both of these processes are characteristic of multifunctional proteins such as homeodomain transcription factors. We conclude that the COAA model can explain two widely recognized features of transcription factor proteins: conserved domains and a tendency to accumulate homopeptides. PMID- 20024540 TI - Coordinate loss of MHC class II expression in the diffuse large B cell lymphoma cell line OCI-Ly2 is due to a novel mutation in RFX-AP. AB - Loss of major histocompatibility complex class II (MHCII) antigen expression on diffuse large B cell lymphoma (DLBCL) corresponds closely with significant decreases in patient survival. However, the mechanisms accounting for MHCII loss in DLBCL have not been thoroughly characterized to date. In this report, we demonstrate that coordinate loss of MHCII expression in OCI-Ly2 DLBCL cells is associated with an 11-base deletion in the cDNA encoding RFX-AP, one of the subunits of the heterotrimeric regulatory factor X (RFX) that is required for activating MHCII transcription. This deletion results in a frameshift in the RFX AP protein beginning at amino acid 234 and, therefore, in the loss of C-terminal amino acids that are required for function. Stable transfection of OCI-Ly2 DLBCL cells with an expression vector for wild-type RFX-AP restores MHCII expression, which strongly suggests that the defect in RFX-AP accounts for MHCII loss in these cells. PMID- 20024541 TI - Synthesis and production of polyhydroxyalkanoates by halophiles: current potential and future prospects. AB - Biodegradable materials with plastic or elastomeric properties are in great demand for a variety of applications. Polyhydroxyalkanoates (PHAs), polyesters synthesized by microorganisms, possess such desired features. Industrial production of PHAs is currently achieved using recombinant Escherichia coli. Nevertheless, recent research on halophiles, salt requiring microorganisms, has shown a remarkable potential for biotechnological production of PHAs. The halophilic archaeon Haloferax mediterranei accumulates a co-polymer, i.e., poly(3 hydroxybutyrate-co-3-hydroxyvalerate) in large amounts using glucose, starch, and hydrolyzed whey as carbon sources. Chemical composition and molecular weight of PHAs produced by H. mediterranei can be modified depending on the substrate utilized as precursor. Phylogenetic studies on haloarchaeal enzymes able to polymerize the components of PHAs (i.e., PHA synthases) reveal a novel cluster, with a close relationship with PHA polymerases of bacteria and archaea found in marine-related niches. On the other hand, sequences of PHA synthases of two halophilic bacteria are more closely affiliated to synthases of Proteobacteria. Several bacterial species of the family Halomonadaceae accumulate PHAs. Halomonas boliviensis reached PHA yields and volumetric productivities close to the highest reported so far. Furthermore, H. boliviensis and other Halomonas species are able to co-produce PHA and osmolytes, i.e., ectoines and hydroxyectoine, in one process. PMID- 20024542 TI - Production of pig liver esterase in batch fermentation of E. coli Origami. AB - The establishment of a fermentation process for the production of pig liver esterase (PLE) in high yields is necessary for industrial applications. In our previous studies, we reported the recombinant expression of PLE in Escherichia coli Origami (DE3) in shake flask. Only a coexpression with chaperones GroEL/ES allowed the production of soluble and active enzyme. The optimization of the cultivation conditions, such as temperature, inducer concentrations, or media compositions to increase enzyme yield in a fermentation process is described here. Using fed-batch fermentation cell densities up to OD = 50 were obtained, but almost no active enzyme was expressed. Only batch fermentation was found suitable for production of active pig liver esterase and cell densities between OD = 7-13 and activities of 300-400 U L(-1) for isoenzyme PLE-1 (gammaPLE) and 1,400 U L(-1) for PLE-5 were obtained after 22 h total cultivation time or 18 h after induction of PLE expression, respectively. PMID- 20024543 TI - Rapid solubilization of insoluble phosphate by a novel environmental stress tolerant Burkholderia vietnamiensis M6 isolated from ginseng rhizospheric soil. AB - We isolated and characterized novel insoluble phosphate (P)-solubilizing bacteria tolerant to environmental factors like high salt, low and high pHs, and low temperature. A bacterium M6 was isolated from a ginseng rhizospheric soil and confirmed to belong to Burkholderia vietnamiensis by BIOLOG system and 16S rRNA gene analysis. The optimal cultural conditions for the solubilization of P were 2.5% (w/v) glucose, 0.015% (w/v) urea, and 0.4% (w/v) MgCl(2).6H(2)O along with initial pH 7.0 at 35 degrees C. High-performance liquid chromatography analysis showed that B. vietnamiensis M6 produced gluconic and 2-ketogluconic acids. During the culture, the pH was reduced with increase in gluconic acid concentration and was inversely correlated with P solubilization. Insoluble P solubilization in the optimal medium was about 902 mg l(-1), which was approximately 1.6-fold higher than the yield in NBRIP medium (580 mg l(-1)). B. vietnamiensis M6 showed resistance against different environmental stresses like 10-45 degrees C, 1-5% (w/v) salt, and 2-11 pH range. The maximal concentration of soluble P produced by B. vietnamiensis M6 from Ca(3)(PO(4))(2), CaHPO(4), and hydroxyapatite was 1,039, 2,132, and 1,754 mg l(-1), respectively. However, the strain M6 produced soluble P with 20 mg l(-1) from FePO(4) after 2 days and 100 mg l(-1) from AlPO(4) after 6 days, respectively. Our results indicate that B. vietnamiensis M6 could be a potential candidate for the development of biofertilizer applicable to environmentally stressed soil. PMID- 20024544 TI - Method to detect only viable cells in microbial ecology. AB - Propidium monoazide can limit the analysis of microbial communities derived from genetic fingerprints to viable cells with intact cell membranes. However, PMA treatment cannot completely suppress polymerase chain reaction (PCR) amplification when the targeted gene is too short. PMA treatment in combination with two-step nested PCR was designed to overcome this problem. Four experiments were performed to determine the limitation of PMA treatment and to evaluate the suitability of the method by applying the following samples: (1) pure cultures of Escherichia coli O157:H7, Enterobacter aerogenes, and Alcaligenes faecalis; (2) pond water samples spiked with heat-killed E. coli O157:H7 and E. aerogenes; (3) anaerobic sludge samples exposed to increasing heat stress; and (4) selected natural samples of estuarine sediment and lake mud. Results from the first two experiments show that PMA treatment cannot efficiently suppress dead cells from PCR amplification when the targeted gene is as short as 190 bp, however, the two step nested PCR can overcome this problem. The last two experiments indicate the method that PMA treatment in combination with two-step nested PCR is useful for viable cells detection in microbial ecology. PMID- 20024545 TI - Loss of phosphomannomutase activity enhances actinorhodin production in Streptomyces coelicolor. AB - Phosphomannomutase (ManB), whose main function is the conversion of mannose-6 phosphate to mannose-1-phosphate, is involved in biosynthesis of GDP-mannose for numerous processes such as synthesis of structural carbohydrates, production of alginates and ascorbic acid, and post-translational modification of proteins in prokaryotes and eukaryotes. ManB isolated from Streptomyces coelicolor was shown to have both phosphomannomutase and phosphoglucomutase activities. Deletion of manB in S. coelicolor caused a dramatic increase in actinorhodin (ACT) production in the low-glucose Difco nutrient (DN) medium, whereas the wild-type strain did not produce ACT on this medium. Experiments involving complementation of the manB deletion showed that increased ACT production in DN media was due to blockage of phosphomannomutase activity rather than phosphoglucomutase activity. This result therefore provides useful information for the design of strategies that enhance antibiotic production through the control of carbon flux. PMID- 20024546 TI - New applications and performance of bioelectrochemical systems. AB - Bioelectrochemical systems (BESs) are emerging technologies which use microorganisms to catalyze the reactions at the anode and/or cathode. BES research is advancing rapidly, and a whole range of applications using different electron donors and acceptors has already been developed. In this mini review, we focus on technological aspects of the expanding application of BESs. We will analyze the anode and cathode half-reactions in terms of their standard and actual potential and report the overpotentials of these half-reactions by comparing the reported potentials with their theoretical potentials. When combining anodes with cathodes in a BES, new bottlenecks and opportunities arise. For application of BESs, it is crucial to lower the internal energy losses and increase productivity at the same time. Membranes are a crucial element to obtain high efficiencies and pure products but increase the internal resistance of BESs. The comparison between production of fuels and chemicals in BESs and in present production processes should gain more attention in future BES research. By making this comparison, it will become clear if the scope of BESs can and should be further developed into the field of biorefineries. PMID- 20024547 TI - Reduced immune effector cell NKG2D expression and increased levels of soluble NKG2D ligands in multiple myeloma may not be causally linked. AB - BACKGROUND: There is limited understanding of the dysregulation of the innate immune system in multiple myeloma (MM). We analysed the expression of the activating receptor NKG2D on NK cells and T cells of MM patients and investigated the impact of soluble versus membrane-bound NKG2D ligands on the expression of NKG2D. DESIGN: NKG2D expression on NK cells and CD8+ alphabeta T cells from patients with MM or monoclonal gammopathy of uncertain significance and healthy controls was examined flow-cytometrically. Sera from patients and controls were analysed for soluble NKG2D ligands (sNKG2D ligands). RESULTS: Significantly fewer NK cells and CD8+ alphabeta T cells from patients expressed NKG2D compared to healthy controls (NK cells: median 54% interquartile range (IQR) 32-68 versus 71% IQR 44-82%, P = 0.017, CD8+ alphabeta T cells: median 63% IQR 52-81 versus 77% IQR 71-90%, P = 0.018). The sNKG2D ligand sMICA was increased in patients [median 175 (IQR 87-295) pg/ml] versus controls [median 80 (IQR 32-129) pg/ml, P < 0.001], but levels of sMICA did not correlate with NKG2D expression on effector cells. To elucidate the mechanism of NKG2D down-regulation, we incubated lymphocytes from healthy donors in the presence of sNKG2D ligands or in co culture with MM cell lines. sNKG2D ligands in clinically relevant concentrations did not down-regulate NKG2D expression, but co-culture of effector cells with myeloma cells with high surface expression of NKG2D ligands reduced NKG2D expression significantly. CONCLUSIONS: These results indicate that MM is associated with a significant reduction in NKG2D expression which may be contact mediated rather than caused by soluble NKG2D ligands. PMID- 20024548 TI - A case of pentacuspid pulmonary valve. AB - A pentacuspid pulmonary valve is a rare congenital anomaly. The present report describes a case of pentacuspid pulmonary valve encountered during the autopsy of a 50-year-old man with a ruptured abdominal aortic aneurysm. The pentacuspid pulmonary valve had three relatively equal cusps and two smaller cusps. In this case, a tricuspid aortic valve was identified with normal structure and no evidence of heart failure was found clinically or pathologically. PMID- 20024549 TI - Imatinib, cytokines and interstitial lung disease in a patient with primary myelofibrosis. PMID- 20024550 TI - Acquired Hb H disease associated with elevated Hb F level in patient affected by primary myelofibrosis. PMID- 20024551 TI - Pulmonary marginal zone B-cell lymphoma of MALT type--what is a prognostic factor and which is the optimal treatment, operation, or chemotherapy?: Consortium for Improving Survival of Lymphoma (CISL) study. AB - Pulmonary marginal zone B-cell lymphoma of the MALT type (P-MZL) is a relatively rare form of lymphoma. We conducted a retrospective analysis of the clinical features and treatment outcomes of P-MZL for the evaluation of prognostic factors, and to collect information about the optimal treatment modality for this condition. From 1991 to 2008, a total of 61 patients with biopsy-confirmed P-MZL were retrospectively analyzed. The median age of our subjects was 60 (range, 34 79) years. Twenty-five of the patients (41%) were initially diagnosed without any symptoms. Video-assisted thoracic surgery was utilized for diagnosis in 19 patients (31%). Thirty-eight patients' conditions (62%) involved a single lobe. Lung lesions were bilateral in 15 patients (25%). Eleven patients evidenced synchronous involvement of extra-pulmonary site MZL. Overall, 56 of 61 patients were treated with surgery (n = 22), chemotherapy (n = 28), or radiotherapy (n = 6). Among them, 46 patients achieved complete or partial remission. The median time to progression (TTP) was 5.6 (95% CI, 2.6-8.6) years. Five patients died during follow-up. Extra-pulmonary MZL and LN involvement were shown to be poor prognostic factors for TTP. We noted no differences between the operation group and chemotherapy group in terms of TTP. P-MZL tends to be an indolent disease characterized by prolonged survival with frequent relapses. This is similar to what is observed with other cases of MALT-type site MZL. In order to conserve lung function and reduce the risks of operation, chemotherapy should be considered as a first-line option for the treatment of P-MZL. PMID- 20024552 TI - An aggressive and lethal course of Churg-Strauss syndrome with alveolar hemorrhage, intestinal perforation, cardiac failure and peripheral neuropathy. AB - Churg-Strauss syndrome (CSS) is a rare type of necrotizing vasculitis affecting small to medium-sized vessels typically characterized by asthma, lung infiltrates, necrotizing granulomas and hypereosinophilia. Herein, we describe a case of CSS presenting severe and aggressive course. A 35-year-old male patient with weight loss, dyspepsia, dyspnea and hemoptysis was admitted. The laboratory analyses indicated a remarkable eosinophilia, elevated levels of serum total IgE and positive cANCA. Thorax CT findings were suggestive of alveolar hemorrhage. Bronchoalveolar lavage revealed alveolar hemorrhage with eosinophilia and transbronchial lung biopsy showed eosinophilic vasculitis. Cardiac enzymes were increased and murmurs were audible revealing cardiomyopathy proven by echocardiography. Pulse cyclophosphamide and methyl prednisolone was immediately started. On the 21st day, intestinal perforation developed and urgent surgery was performed. During a follow-up, although a radiological improvement was observed in the chest X-ray, cardiac failure, peripheral neuropathy and skin lesions developed and high-dose intravenous immunoglobulin and anti-TNF therapy (adalimumab) were applied. Despite the therapy, he died from heart failure and septicemia at 68th day of therapy. PMID- 20024553 TI - Primary central nervous system vasculitis: clinical experiences with 21 new European cases. AB - Primary central nervous system vasculitis (PCNSV) represents a rare and poorly understood form of vascular inflammatory disease restricted to the brain and spinal cord. This study was undertaken to better characterize demographic and clinical features and to evaluate treatments and outcome. Our cohort included 21 white patients with PCNSV treated in a single German institution between 2003 and 2008. Clinical and diagnostic features were obtained by retrospective chart review; follow-up information and outcome were obtained prospectively. Clinical findings and outcomes were compared among patients categorized by different characteristics. The mean age at onset of symptoms was 42.48 years. In 6 (28.6%) of 21 patients, PCNSV diagnosis was established based on positive CNS tissue histopathology. Additional angiography was only performed in two of these six patients and was negative in both cases. For 13 patients (61.9%), the diagnosis was confirmed by positive cerebral angiogram. The most frequent clinical presentations over the whole follow-up period were hemiparesis (76.2%), cerebral ischemia (66.7%), headache (57.1%) and altered cognition (52.4%). Patients with MR-tomographic evident microangiopathy showed significantly worse modified Rankin disability scores during the onset and at last visit compared to the other patients (P = 0.047, P = 0.039). Clinical features and course of PCNSV of Europeans analyzed in this German study are comparable to American results but some further reported characteristics of different subtypes could not be duplicated. Although this well-defined analysis adds to our knowledge of PCNSV, it also raises important open questions. The enigma of PCNSV could only be solved by implementing large prospective multicenter databases and elucidating animal models. PMID- 20024554 TI - Effects of the anti-interleukin-6 receptor antibody, tocilizumab, on serum lipid levels in patients with rheumatoid arthritis. AB - We investigated the effects of anti-IL-6 receptor antibody, tocilizumab (TCZ), on lipid metabolism. Nineteen patients with rheumatoid arthritis (RA), entered in clinical case-control study of SAMURAI trial at Sasebo Chuo Hospital, were examined. Nine patients received TCZ monotherapy at 8 mg/kg intravenously every 4 weeks (TCZ group) and 10 patients received conventional DMARDs (control group). Serum total cholesterol (TC), high-density lipoprotein cholesterol (HDL), low density lipoprotein cholesterol (LDL), apolipoprotein (Apo) A-1, Apo A-2 and Apo B as well as disease activity score (DAS), C-reactive protein and serum amyloid A protein were examined at baseline and after 3 months of the treatment. IL-6 inversely was correlated with LDL, Apo A-1 and Apo A-2, and also tended to correlate with Apo B. In TCZ group, serum levels of TC, HDL, LDL, Apo A-1 and Apo A-2 were significantly increased after 3 months treatment with TCZ. There was no significant change in Apo B, the atherogenic index, and TC/HDL by the TCZ treatment. Changes in the DAS28-ESR negatively correlated with those in TC. In one patient, whose serum level of TCZ was not detected after 3 months of the treatment, the absence of the increment in serum levels of Apo A-1 and A-2 in the patient was remarkable. All of the markers did not change during 3 months in control group. These data may raise an important issue to evaluate the impact of these alternations in lipid metabolism for longer periods in RA patients treated with TCZ. PMID- 20024555 TI - Serum chemokines in patients with rheumatoid arthritis treated with etanercept. AB - Chemokines promote leucocyte traffic into the synovium, leading to the initiation and progression of the rheumatoid arthritis (RA). The aim of the study was to determine the effects of etanercept, a soluble tumour necrosis factor receptor (sTNFr), on the serum chemokines levels in patients with active RA. Patients were treated with 50 mg of subcutaneous injection of etanercept per week and methotrexate (10-25 mg/week). Serum levels of interleukin-8 (IL-8), RANTES (regulated upon activation, normal T cell expressed and secreted) and monocyte chemoattractant protein-1 (MCP-1) were assessed by ELISA at months 0, 3, 6, 9 and 12, prior to injection. 3-month treatment with etanercept diminished serum concentrations of IL-8, RANTES and MCP-1 (P < 0.05, P < 0.01 and P < 0.001, respectively). Subsequent etanercept administrations prolonged decrease in serum chemokines levels and in the case of IL-8 even intensified the reduction of its concentration in serum. These changes were accompanied by significant decrease of disease activity score (DAS28) (in all cases P < 0.001). Prior to the first etanercept administration, serum concentrations of studied chemokines correlated with markers of RA activity such as the erythrocyte sedimentation rate (ESR) and DAS28. Following next drug injection such associations were less or not significant. Therapy with etanercept and MTX not only caused a clinical improvement but also diminished serum chemokines levels in RA patients. Further treatment with etanercept sustained chemokines suppression. PMID- 20024556 TI - Polymorphism of the extrapituitary prolactin promoter and systemic sclerosis. PMID- 20024557 TI - C1-C2 spondylodiscitis in an adult with SAPHO syndrome: an unusual presentation. AB - The main advantage of recognition and diagnosis of SAPHO syndrome is the avoidance of unnecessary prolonged antibiotic treatment and repeated invasive procedures. The combination of synovitis, acne, pustulosis, hyperostosis and aseptic osteitis is known as SAPHO syndrome. The most common site of the disease is the upper anterior chest wall, characterized by predominantly osteosclerotic lesions, hyperostosis, and arthritis of the adjacent joints. Osteosclerosis of the vertebral bodies, hyperostosis, and erosions of the vertebral plates are rarely encountered. We present a case of SAPHO syndrome with C1-C2 spondylodiscitis. Diagnosis of SAPHO syndrome was established using Khan et al. criteria (Schilling, SAPHO syndrome, Encyclopedie Orphanet, 2004). CT showed osteosclerotic lesions of dens axis. MRI sequences reveal inflammatory bone marrow oedema. Radiologists should be aware of this unusual syndrome to avoid misdiagnosis (tumour/infection), unnecessary surgery, and antibiotic therapy. PMID- 20024558 TI - Clopidogrel-associated acute arthritis. AB - A 54-year-old Japanese man was urgently admitted to our hospital because of "unmovable" polyarthralgia, high fever and pruritus. He had taken clopidogrel 3 weeks before this admission to prevent ischemic cerebrovascular events. On the laboratory values, acute phase reactants and serum IgE were elevated. Although his symptoms had promptly improved with non-steroidal anti-inflammatory drugs (NSAIDs) and cessation of the drug, the same phenomena were re-induced by re administration of the drug. According to the above manifestations, he was diagnosed as suffering from clopidogrel-associated acute arthritis. PMID- 20024559 TI - [Femtosecond laser-assisted penetrating keratoplasty]. AB - The femtosecond laser technique allows completely new trephination methods in penetrating keratoplasty with profiles in the graft and the host cornea. The most common so-called profiled trephinations are the top hat and mushroom profiles. Due to the profile it is easier to get a watertight wound closure intraoperatively and due to the larger wound surface the wound healing is faster and more stable. This will possibly allow an earlier suture removal with final visual rehabilitation. First clinical results show that femtosecond laser assisted penetrating keratoplasty is a safe surgical procedure. The long term results after complete suture removal will show whether the astigmatism results are better than those of conventional trephination techniques. PMID- 20024560 TI - [Perioperative conversion of oral anticoagulants to heparin (bridging) in ophthalmic medicine]. AB - Since topical anesthesia was introduced in ophthalmic surgery, anticoagulation therapy can often be used in patients with thromboembolic risk. But some surgical procedures with an increased risk for intraoperative bleeding necessitate changing a patient's Coumadin therapy to heparin. To reduce intraoperative bleeding and perioperative thromboembolic complications, ophthalmologists and referring general practitioners should cooperate closely. PMID- 20024561 TI - [Visual diagnosis: Waardenburg syndrome]. AB - BACKGROUND: Waardenburg syndrome (WS) is a rare disease characterized by a sensorineural hearing loss and pigment anomalies of the iris, skin and hair due to mutations in PAX3. WS can be subdivided into four groups according to major and minor clinical signs. CASE REPORT: We report the case of a 2 1/2-year-old coloured patient who presented in our department of paediatric ophthalmology for a syndrome search. The patient presented with hearing loss, brilliant blue iris colour and dystopia canthorum. The patient was slightly hypermetropic. Visual acuity was within normal limits according to the Cardiff acuity test. The ocular fundus examination revealed no abnormalities. CONCLUSION: According to the major and minor criteria defined by the Waardenburg consortium our patient showed the major criteria of WS1, i.e. hearing loss, hypopigmentation of the pigment epithelium of the iris and dystopic canthi. Diagnosis of WS is usually based on the clinical presentation. An additional molecular genetic analysis is possible. PMID- 20024562 TI - [Morbus Morbihan : A rare cause of edematous swelling of the eyelids]. AB - Morbus Morbihan is a rare disease characterized by persistent erythema and edema of the central and upper half of the face, mainly restricted to the forehead, glabella, eyelids, nose and cheeks. It is thought to be a rare complication of rosacea and may lead to fibrotic induration. The characteristic features are a typical clinical picture with histological alterations. Diagnosis and treatment are difficult because of the absence of specific laboratory findings and in most cases resistance to clinical therapy.We report the first case of morbus Morbihan in the ophthalmic literature. A 53-year-old woman complained of erythematous and edematous eyelids which caused a significant visual field defect for 4 years. The typical clinical picture, histological findings and the exclusion of several differential diagnoses led to the diagnosis of morbus Morbihan. Oral rosacea treatment did not result in any improvement, therefore, eyelid reduction surgery followed by lymphatic drainage was performed. Apart from cosmetic improvement a significant reduction of visual field defects was observed postoperatively. PMID- 20024563 TI - [Solitary synchronous intraocular metastasis of a peripheral non-small cell lung cancer : A multidisciplinary approach with curative intention]. AB - A solitary synchronous distant metastasis of newly diagnosed non-small cell lung cancer (NSCLC) occurs in only a small number of patients. The available literature suggests a significant rise in long-term survival for these patients when resection of the bronchial carcinoma and the metastasis are feasible and performed. To demonstrate a multidisciplinary concept, we present the case of a patient with a peripheral NSCLC and a solitary synchronous intraocular metastasis. To our knowledge, this concept with curative intention has not been published before regarding a patient with an intraocular metastasis. PMID- 20024564 TI - [Laser embolectomy for central retinal artery occlusion]. AB - We describe a case of successful Nd:YAG laser embolectomy in a patient with occlusions of the retinal artery affecting the macula and reducing visual acuity. The treatment can only be recommended in cases with a visible embolus and short duration of symptoms. PMID- 20024565 TI - [Value-based medicine in ophthalmology]. PMID- 20024566 TI - [Patient-reported and patient-weighted outcomes in ophthalmology]. AB - Considering patients' values and preferences in comparative effectiveness research (CER) is one of the main challenges in ophthalmology (value-based medicine). This article defines core terms in CER. The concept of patient relevant (or patient-important) outcomes is distinguished from patient-reported outcomes (PRO) by means of examples in the field of ophthalmology. In order to be able to give a consistant recommendation if an intervention leads to conflicting results for different outcomes (trade-off), a ranking of outcomes will be necessary. Examples of studies in glaucoma patients are provided that demonstrate the possibilities of ranking of outcomes based on patient preferences. PMID- 20024567 TI - Intestinal osmoregulatory acclimation and nitrogen metabolism in juveniles of the freshwater marble goby exposed to seawater. AB - The objective of this study was to elucidate the role of the intestine from juveniles of the marble goby, Oxyeleotris marmorata, during seawater (SW) exposure. It has been reported elsewhere that SW-exposed juvenile O. marmorata exhibits hypoosmotic and hypoionic regulation, with the induction of branchial Na(+)/K(+)-ATPase (NKA), Na(+):K(+):2Cl(-) cotransporter (NKCC), and cystic fibrosis transmembrane receptor-like chloride channels. Here, we report that SW exposure also led to significant increases in the activity and protein abundance of NKA in, and probably an increase in Na(+) uptake through, its intestine. Additionally, there was an increase in apical NKCC immunoreactivity in the intestinal epithelium, indicating that there could be increased Cl(-) uptake through the intestine. These results suggest that absorption of ions, and hence water, from the intestinal lumen could be an essential part of the osmoregulatory process in juvenile O. marmorata during exposure to SW. Furthermore, there were significant increases in the glutamate content, and the aminating activity and protein abundance of glutamate dehydrogenase (GDH) in the intestine of fish exposed to SW. Since the intestinal glutamine synthetase activity and protein abundance decreased significantly, and the intestinal glutamine content remained unchanged, in the SW-exposed fish, excess glutamate formed via increased GDH activity in the intestine could be channeled to other organs to facilitate the increased synthesis of amino acids. Taken together, our results indicate for the first time that, besides absorbing ions and water during SW exposure, the intestine of juvenile O. marmorata also participated in altered nitrogen metabolism in response to salinity changes. PMID- 20024568 TI - Minimal invasive fixation of proximal humeral fractures with an intramedullary nail: good results in elderly patients. AB - OBJECTIVE: To report on the results of a minimally invasive technique for the fixation of displaced proximal humeral fractures with a locked intramedullary nail. PATIENTS AND METHODS: All consecutive patients treated with a T2 intramedullary nail between 2004 and 2007 were evaluated. Thirty-three patients were included [mean age 78, m:f ratio (1:4)]. Fracture characteristics were classified according to AO and Neer (eighteen 2-part, eleven 3-part, five 4-part fractures). RESULTS: Functional outcome (Constant Score) was excellent in nine, satisfactory in eight and poor in three patients. Subjective outcome was satisfactory to good for patients with 2-part and 3-part fractures but poor for 4 part fractures. Major complications comprised four fixation failures, two cases of impingement and one deep infection. CONCLUSIONS: Minimally invasive fixation of displaced 2-part and 3-part humeral fractures in an elderly population shows satisfactory to excellent results in 80% of patients. PMID- 20024569 TI - Successful photodynamic therapy with topical 5-aminolevulinic acid for five cases of cervical intraepithelial neoplasia. AB - OBJECTIVES: Photodynamic therapy (PDT) is a minimally invasive treatment for cervical intraepithelial neoplasia. The purpose of this study was to assess the feasibility and effectiveness of PDT in patients with CIN and high-risk HPV infection. METHODS: Five patients diagnosed CIN 2 or CIN 3 with human papillomavirus (HPV) infection were included. Each patient had gynecologic examination including cervical cytology, HPV DNA testing, colposcopy and biopsy. Two grams of 5-aminolevulinic acid (ALA) gel (118 mg/g) was topically applied to the cervix and covered with a special plastic cap for 3-4 h, followed by 20 min illumination of both ecto- and endo-cervical canal with red coherent light (wavelength 633 nm) using a PDT laser and a special light catheter. The PDT therapy was repeated with an interval of 1 week. Follow-up examination including biopsy and histology, colposcopy, HPV DNA testing were carried out after 3, 6 and 9 months. RESULTS: Treatment could be accomplished in all cases and no severe side effect was encountered. All the CIN2 patients had a complete response for 9 months and one CIN3 HPV remained positive for 6 months after three or four treatments. CONCLUSION: PDT seems to be a non-invasive, repeatable procedure for CIN and cervical HPV infection with minimal side effects and can be easily performed on outpatient basis. PMID- 20024570 TI - Longitudinal analysis of changes in energy intake and macronutrient composition during pregnancy and 6 weeks post-partum. AB - PURPOSE: This prospective study was carried out in order to investigate changes in nutritional intake during pregnancy with regard to caloric intake and macronutrient composition in normal-weight pregnant women. METHODS: Using food scales, 32 healthy pregnant women estimated their food intake over a period of 2 days at gestational week 16, 22, 30, 36 and, in addition, 6 weeks after delivery. The recorded food logs were analyzed with respect to caloric intake and macronutrient composition. RESULTS: The women's age was averaged 29.6 +/- 4.5 years, and the mean pre-pregnancy BMI was 22.4 +/- 2.5 kg/m2. Mean weight gain was 16.5 +/- 4.5 kg. On average, birth took place during the 40th week of gestation, with the babies having a mean birth weight of 3,515 +/- 406 g. Caloric intake during the gestation period and post-partum showed no statistically significant changes and added up 9,237 +/- 1,876 kJ/day during the 16th week of gestation, 9,496 +/- 2,437 kJ/day during the 22nd, 9,073 +/- 1,863 kJ/day during the 30th, 9,525 +/- 2,135 kJ/day during the 36th week and 8,445 +/- 2,160 kJ/day 6 weeks after delivery. No significant changes were observed in the composition of macronutrients during the pregnancy. The daily composition of macronutrients was as follows: carbohydrates 281 +/- 57 g/day (51.7%), fat 86 +/- 16 g/day (35.8%) and protein 75 +/- 13 g/day (13.9%). However, we observed a decrease in carbohydrate intake of 239 +/- 72 g/day (48%) after birth. CONCLUSIONS: Healthy pregnant women showed no significant changes in their caloric intake or nutritional profiles. Performed with an accurate measurement, this prospective study shows that healthy pregnant require neither increased caloric intake nor a change in macronutrient composition. PMID- 20024571 TI - Is single umbilical artery an independent risk factor for perinatal mortality? AB - OBJECTIVE: To evaluate perinatal outcome of fetuses with isolated single umbilical artery (SUA), and specifically to examine whether an isolated SUA is an independent risk factor for perinatal mortality. METHODS: A population-based study was conducted, comparing pregnancies of women with and without SUA. Deliveries occurred between the years 1988-2006, in a tertiary medical center. Multiple gestations, chromosomal abnormalities and malformations were excluded from the analysis. Stratified analysis was performed using multiple logistic regression models to evaluate the association between SUA and perinatal mortality, while controlling for confounders. RESULTS: Out of 194,809 deliveries, 243 (0.1%) were of fetuses with isolated SUA. Fetuses with SUA were smaller (2,844 +/- 733 vs. 3,197 +/- 530 g, P < 0.001), and were delivered at an earlier gestational age (38.3 +/- 3.0 vs. 39.3 +/- 2.1 weeks, P < 0.001), when compared with fetuses with normal umbilical vessels. Mothers to fetuses with isolated SUA tended to have a history of infertility treatments (4.5 vs. 1.7%; P = 0.001) when compared with the comparison group. Fetuses with SUA had more complications, including fetal growth restriction (FGR 9.5 vs. 1.9%, P < 0.001), polyhydramnios (11.5 vs. 3.7%; P < 0.001) and oligohydramnios (6.6 vs. 2.2%; P < 0.001). Deliveries of SUA fetuses had higher rates of placental abruption (3.3 vs. 0.7%; P < 0.001), placenta previa (1.2 vs. 0.4%; P = 0.03) and cord prolapse (2.9 vs. 0.4%; P < 0.001). Higher rates of cesarean deliveries were noted in this group (23.9 vs. 12.2%; P < 0.001). SUA newborns had higher rates of low Apgar scores (<7) in one (11.8 vs. 3.7%; P < 0.001) and 5 min (3.5 vs. 0.4%; P < 0.001). Higher rates of perinatal mortality were noted in the SUA group, as compared to fetuses with normal umbilical vessels (6.6 vs. 0.9%, OR 7.78; 95% CI 4.7-13.0; P < 0.001). Using a multiple logistic regression model, controlling for possible confounders, such as FGR, oligohydramnios, polyhydramnios, prolapse of cord, maternal hypertension and diabetes mellitus, isolated SUA remained an independent risk factor for perinatal mortality (adjusted OR = 3.91, 95% CI 2.06-7.43; P < 0.001). CONCLUSION: Isolated SUA in our population was noted as an independent risk factor for perinatal mortality. PMID- 20024572 TI - Laparoscopic subtotal hysterectomy for large uteri using modified five port technique. AB - OBJECTIVE: To evaluate outcome of laparoscopic subtotal hysterectomy (LASH) performed in women with large uteri using a modified five port technique. DESIGN: Prospective Cohort Study (Canadian Task Force classification III). SETTING: Advanced laparoscopic Unit at the Whipps Cross University Hospital, London, UK. INCLUSION CRITERIA: (1) All women with symptomatic and large uteri undergoing LASH, (2) previous three normal cervical smears and consent for future smears as protocol, (3) patient consent for the procedure. METHODS: Prospectively collected database was analysed statistically for 29 women with enlarged uteri weighing more than 300 g who underwent LASH using the modified five port technique, during the time period of 24 months from Jan 2007 to Jan 2009. This technique demonstrates that the port placement facilitates the procedure. RESULTS: The mean age of the patients was 47 +/- 4 years. Median BMI was 29 (range 22-41), Most common indication of surgery was menorrhagia in 90% of cases followed by palpable lump in 67% of cases. Median size of uterus was 18 weeks (range 12-30). Mean operative time was 84 +/- 42 min. Median blood loss was 200 ml (range 100-800 ml).There was only one case with major complication such as bladder injury which was diagnosed and repaired intraoperatively. Mean hospital stay was 2 +/- 1 day. None of the cases needed conversion to laparotomy. CONCLUSION: Modified five port technique of LASH for large uteri provides optimal exposure of the operative field to increase the efficiency and ease of the procedure and has its added cosmetic benefits. PMID- 20024573 TI - Risk factors in prolonged postpartum urinary retention: an analysis of six cases. AB - PURPOSE: Prolonged first and second stage of labor, isolated prolongation of the second stage, forceps delivery or vacuum extraction, perineal laceration, nulliparity and epidural anesthesia are known risk factors for developing prolonged postpartum urinary retention (PUR). The aim of our study was to analyze number and constellations of these risk factors, in prolonged postpartum urinary retention (PPUR) in our own unit to facilitate the identification of patients at high risk and thus to prevent bladder overdistension by early intervention. METHODS: We performed a retrospective analysis of all our cases with PPUR between 2003 and 2008 including variables like age weight, height, body mass index, fetal birth weight and head circumference. RESULTS: The incidence of PPUR at our institution is low being 0.06%. No woman combined all six risk factors. The majority had five risk factors, all had at least four. An isolated prolonged second stage of labor was common to all patients with PPUR. Five women had an epidural anesthesia, three were nulliparous and only two women delivered spontaneously. All but one woman suffered from perineal tears. Interestingly, fetal head circumference was larger than 36 cm in four of six cases. CONCLUSION: In contrast to simple PUR, the prolonged form of PUR could be the result of a cumulative effect of different single risk factors. PMID- 20024574 TI - Parkinsonism in a population of northern Tanzania: a community-based door-to-door study in combination with a prospective hospital-based evaluation. AB - The prevalence of Parkinson's disease (PD) in sub-Saharan Africa is still a matter of debate. The few studies that have been conducted have shown prevalences lower than those in the western world. Whether this represents a genuine finding is unclear to date. In northern Tanzania, we have conducted a hospital-based evaluation and a community-based door-to-door study to assess the prevalence of Parkinsonism, including that of PD. Over a period of 8 months, all patients admitted to a mid-size rural hospital were screened for PD. In parallel, 1,569 people aged >or=50 years were recruited from the communities and assessed for PD with standard questions. Sampling was performed according to the method of "multistage cluster sampling." The questions had previously been tested in a pilot study prior to the survey. People who screened positive were examined by a specialist neurologist. In the hospital, eight of 740 people with neurological diagnoses had Parkinsonism, of whom three patients had a diagnosis of PD. In the community-based study, 18 people answered positively to least one of the 12 screening questions. However, the diagnosis of PD could not be confirmed by further examination in any of them. The prevalence of PD in northern Tanzania was found to be very low. This result would need confirmation in studies with larger populations, ideally of different African ethnicities. PMID- 20024575 TI - Men and women exhibit a similar time to task failure for a sustained, submaximal elbow extensor contraction. AB - Sex differences in muscle fatigue-resistance have been observed in a variety of muscles and under several conditions. This study compared the time to task failure (TTF) of a sustained isometric elbow extensor (intensity 15% of maximal strength) contraction in young men (n = 12) and women (n = 11), and examined if their neurophysiologic adjustments to fatigue differed. Motor-evoked potential amplitude (MEP), silent period duration, interference electromyogram (EMG) amplitude, maximal muscle action potential (M (max)), heart rate, and mean arterial pressure were measured at baseline, during the task, and during a 2-min ischemia period. Men and women did not differ in TTF (478.2 +/- 31.9 vs. 500.4 +/ 41.3 s; P = 0.67). We also performed an exploratory post hoc cluster analysis, and classified subjects as low (n = 15) or high endurance (n = 8) based on TTF (415.3 +/- 16.0 vs. 626.7 +/- 25.8 s, respectively). The high-endurance group exhibited a lower MEP and EMG at baseline (MEP 16.3 +/- 4.1 vs. 37.2 +/- 3.0% M (max), P < 0.01; EMG 0.98 +/- 0.18 vs. 1.85 +/- 0.26% M (max), P = 0.03). These findings suggest no sex differences in elbow extensor fatigability, in contrast to observations from other muscle groups. The cluster analyses results indicated that high- and low-endurance groups displayed neurophysiologic differences at baseline (before performing the fatigue task), but that they did not differ in fatigue-induced changes in their neurophysiologic adjustments to the task. PMID- 20024576 TI - Pacing strategy during the initial phase of the run in triathlon: influence on overall performance. AB - The aim of the present study was to determine the best pacing strategy to adopt during the initial phase of a short distance triathlon run for highly trained triathletes. Ten highly trained male triathletes completed an incremental running test to determine maximal oxygen uptake, a 10-km control run at free pace and three individual time-trial triathlons (1.5-km swimming, 40-km cycling, 10-km running) in a randomised order. Swimming and cycling speeds were imposed as identical to the first triathlon performed and the first run kilometre was done alternatively 5% faster (Tri-Run(+5%)), 5% slower (Tri-Run(-5%)) and 10% slower (Tri-Run(-10%)) than the control run (C-Run). The subjects were instructed to finish the 9 remaining kilometres as quickly as possible at a free self-pace. Tri Run(-5%) resulted in a significantly faster overall 10-km performance than Tri Run(+5%) and Tri-Run(-10%) (p < 0.05) but no significant difference was observed with C-Run (p > 0.05) (2,028 +/- 78 s vs. 2,000 +/- 72 s, 2,178 +/- 121 s and 2,087 +/- 88 s, for Tri-Run(-5%), C-Run, Tri-Run(+5%) and Tri-Run(-10%), respectively). Tri-Run(+5%) strategy elicited higher values for oxygen uptake, ventilation, heart rate and blood lactate at the end of the first kilometre than the three other conditions. After 5 and 9.5 km, these values were higher for Tri Run(-5%) (p < 0.05). The present results showed that the running speed achieved during the cycle-to-run transition is crucial for the improvement of the running phase as a whole. Triathletes would benefit to automate a pace 5% slower than their 10-km control running speed as both 5% faster and 10% slower running speeds over the first kilometre involved weaker overall performances. PMID- 20024577 TI - Effect of low-level laser therapy (GaAs 904 nm) in skeletal muscle fatigue and biochemical markers of muscle damage in rats. AB - We wanted to test if pre-exercise muscle irradiation with 904 nm laser affects the development of fatigue, blood lactate levels and creatine kinase (CK) activity in a rat model with tetanic contractions. Thirty male Wistar rats were divided into five groups receiving either one of four different laser doses (0.1, 0.3, 1.0 and 3.0 J) or a no-treatment control group. Laser irradiation was performed immediately before the first contraction for treated groups. Electrical stimulation was used to induce six tetanic tibial anterior muscle contractions with 10 min intervals between them. Contractions were stopped when the muscle force fell to 50% of the peak value for each contraction; blood samples were taken before the first and immediately after the sixth contraction. The relative peak forces for the sixth contraction were significantly better (P < 0.05) in the two laser groups irradiated with highest doses [151.27% (SD +/- 18.82) for 1.0 J, 144.84% (SD +/- 34.47) for 3.0 J and 82.25% (SD +/- 11.69) for the control group]. Similar significant (P < 0.05) increases in mean performed work during the sixth contraction for the 1.0 and 3.0 J groups were also observed. Blood lactate levels were significantly lower (P < 0.05) than the control group in all irradiated groups. All irradiated groups except the 3.0 J group had significantly lower post-exercise CK activity than the control group. We conclude that pre exercise irradiation with a laser dose of 1.0 J and 904 nm wavelength significantly delays muscle fatigue and decreases post-exercise blood lactate and CK in this rat model. PMID- 20024578 TI - Effects of a mental task on splanchnic blood flow in fasting and postprandial conditions. AB - We tested the hypothesis that a mental task attenuates the meal-induced vasodilation in the splanchnic vasculature. Ten subjects performed a 5-min colour word conflict test (CWT) under fasting and postprandial conditions. Subjects in the postprandial condition had ingested solid food with an energy content of 300 kcal (1,255 kJ) before either performing the CWT (mental task trial) or resting (resting control trial). The mean blood velocities (MBV) in the coeliac artery (CA) and superior mesenteric artery (SMA), and the mean arterial pressure (MAP) were measured. The MBV in the CA and SMA were divided by the MAP to assess the vascular conductance (VC). The MBV in the CA and SMA were significantly increased by the CWT under fasting conditions. In the postprandial condition, the MBV in the CA and SMA significantly increased immediately after the meal in both the mental task and resting control trials. The VC in the SMA, but not in the CA, was significantly decreased by the CWT under fasting conditions. In the postprandial conditions, there was no significant difference in the VC in both arteries between mental task and resting control trials. These results suggest that a mental task exerts different effects on the CA and SMA under fasting but not postprandial condition. The vasoconstrictive effect of a mental task on the SMA does not counter the vasodilatory effect of meal ingestion. PMID- 20024579 TI - Are there differences in running economy at different velocities for well-trained distance runners? AB - The present study investigated whether there are differences in running economy at different velocities for well-trained distance runners, and to what extent a commonly used incremental protocol for measuring oxygen uptake (VO(2)) at different velocities affects the reliability of these measurements. Fifteen well trained distance runners (9 male and 6 female) participated in this study. Gross oxygen cost of running (C (R)), heart rate (HR) and [La(-)](b) during 5-min runs at velocities ranging from 8.0 to 17 km h(-1), representing intensities ranging from 60 to 90% of maximal oxygen consumption (VO(2max)) was measured on two different days in random order. The athletes were also tested for lactate threshold, VO(2max) and time to exhaustion at MAS ((t)MAS). No significant differences in C (R) between the different relative velocities or the different set velocities were found up to 90% of VO(2max). The incremental protocol for measuring VO(2) at different velocities was found not to affect the reliability of these measurements. All athletes reached their VO(2max) whilst running to exhaustion at MAS. The females showed significantly lower VO(2max), but significantly better C (R) than the males. At velocities representing intensities between 60 and 90% of VO(2max), no differences in C (R) were found. The commonly used incremental protocol for measuring oxygen uptake (VO(2)) at different velocities was found not to affect the reliability of these measurements. This means that C (R) measured at sub-maximal velocities are representative for C (R) at race velocity for distances above 10,000 m for most runners. PMID- 20024580 TI - The multiple actions of NO. AB - Studies in the last two decades have firmly established that nitric oxide (NO) exerts a broad range of effects on bodily functions including muscle contractility, platelet aggregation, metabolism, neuronal activity, and immune responses. The underlying mechanisms rely primarily on elevating guanosine 3',5' cyclic monophosphate due to the stimulation of soluble guanylyl cyclase, inhibiting mitochondria respiration by the action on cytochrome C oxidase, and nitrosylating proteins and enzymes. Under pathophysiological conditions, an increased production of NO concurrently with an enhanced generation of superoxide leads to the formation of peroxynitrite, a potent oxidative agent, and thus tissue injuries. This article intends to provide a brief review on the effects of NO in the modulations of muscle contractility, platelet aggregation, metabolism, neuronal activity, and immune responses. The actions of NO vary depending on the interactions between this gaseous molecule, its derivates, and their effectors as well as the local redox environments. Considering the complexity of these interactions and the widespread presence of NO in various body systems and cell types, there is no doubt that this area of research will remain very challenging and rewarding in the foreseeable future. PMID- 20024581 TI - Separable functions of wingless in distal and ventral patterning of the Tribolium leg. AB - The gene wingless (wg) in Drosophila is an important factor in leg development. During embryonic development wg is involved in the allocation of the limb primordia. During imaginal disk development wg is involved in distal development and it has a separate role in ventral development. The expression pattern of wg is highly conserved in all arthropods (comprising data from insects, myriapods, crustaceans, and chelicerates), suggesting that its function in leg development is also conserved. However, recent work in other insects (e.g. the milkweed bug Oncopeltus fasciatus) argued against a role of wg in leg development. We have studied the role of wg in leg development of the flour beetle Tribolium castaneum. Using stage-specific staggered embryonic RNAi in wild-type and transgenic EGFP expressing enhancer trap lines we are able to demonstrate separable functions of Tribolium wg in distal and in ventral leg development. The distal role affects all podomeres distal to the coxa, whereas the ventral role is restricted to cells along the ventral midline of the legs. In addition, severe leg defects after injection into early embryonic stages are evidence that wg is also involved in proximal development and limb allocation in Tribolium. Our data suggest that the roles of wg in leg development are highly conserved in the holometabolous insects. Further studies will reveal the degree of conservation in other arthropod groups. PMID- 20024582 TI - Can renal oncocytoma be distinguished from chromophobe renal cell carcinoma by the presence of fibrous capsule? AB - The most important differential diagnosis of chromophobe renal cell carcinoma (CRCC) is renal oncocytoma. Due to overlapping morphological characteristics of renal oncocytoma and CRCC, particularly its eosinophilic variant, making a correct diagnosis can be challenging. To date, no data are available on the presence of the tumor fibrous capsule as a diagnostic feature in differentiating these tumors. The main purpose of this study was to establish the presence and compare the thickness of the tumor fibrous capsule between two tumor groups. A total of 37 tumors--18 cases of CRCC (three eosinophilic and 15 classic) and 19 cases of renal oncocytoma--were analyzed. Four slides of each tumor stained with hematoxylin and eosin were first scanned at low-power magnification (x40) to assess the presence of the capsule. If present, the capsule was measured in three different thickest areas at higher magnification (x200). The mean value of capsule thickness was calculated and taken into consideration. The capsule was present in 12 (66.7%) cases of CRCCs and in only two (10.5%) cases of renal oncocytomas. Statistical analysis showed significant difference between the presence of fibrous capsule in these two observed tumor groups (P = 0.001). Average thickness of capsule in CRCCs was 337.7 microm, and 115.4 microm in renal oncocytomas, but the median was not statistically significant (P = 0.198). Studies with a larger number of cases are needed to conclude if this characteristic could be a low-cost, reliable microscopic feature in differentiating between CRCC and renal oncocytoma. PMID- 20024583 TI - Genome-wide discovery of DNA polymorphism in Brassica rapa. AB - Single nucleotide polymorphisms (SNPs) and/or insertion/deletions (InDels) are frequent sequence variations in the plant genome, which can be developed as molecular markers for genetic studies on crop improvement. The ongoing Brassica rapa genome sequencing project has generated vast amounts of sequence data useful in genetic research. Here, we report a genome-wide survey of DNA polymorphisms in the B. rapa genome based on the 557 bacterial artificial clone sequences of B. rapa ssp. pekinensis cv. Chiifu. We identified and characterized 21,311 SNPs and 6,753 InDels in the gene space of the B. rapa genome by re-sequencing 1,398 sequence-tagged sites (STSs) in eight genotypes. Comparison of our findings with a B. rapa genetic linkage map confirmed that STS loci were distributed randomly over the B. rapa whole genome. In the 1.4 Mb of aligned sequences, mean nucleotide polymorphism and diversity were theta = 0.00890 and pi = 0.00917, respectively. Additionally, the nucleotide diversity in introns was almost three times greater than that in exons, and the frequency of observed InDel was almost 17 times higher in introns than in exons. Information regarding SNPs/InDels obtained here will provide an important resource for genetic studies and breeding programs of B. rapa. PMID- 20024584 TI - Examination of FGFRL1 as a candidate gene for diaphragmatic defects at chromosome 4p16.3 shows that Fgfrl1 null mice have reduced expression of Tpm3, sarcomere genes and Lrtm1 in the diaphragm. AB - Fgfrl1 (also known as Fgfr5; OMIM 605830) homozygous null mice have thin, amuscular diaphragms and die at birth because of diaphragm hypoplasia. FGFRL1 is located at 4p16.3, and this chromosome region can be deleted in patients with congenital diaphragmatic hernia (CDH). We examined FGFRL1 as a candidate gene for the diaphragmatic defects associated with 4p16.3 deletions and re-sequenced this gene in 54 patients with CDH. We confirmed six known coding single nucleotide polymorphisms (SNPs): c.209G > A (p.Pro20Pro), c.977G > A (p.Pro276Pro), c.1040T > C (p.Asp297Asp), c.1234C > A (p.Pro362Gln), c.1420G > T (p.Arg424Leu), and c.1540C > T (p.Pro464Leu), but we did not identify any gene mutations. We genotyped additional CDH patients for four of these six SNPs, including the three non-synonymous SNPs, to make a total of 200 chromosomes, and found that the allele frequency for the four SNPs, did not differ significantly between patients and normal controls (p > or = 0.05). We then used Affymetrix Genechip Mouse Gene 1.0 ST arrays and found eight genes with significantly reduced expression levels in the diaphragms of Fgfrl1 homozygous null mice when compared with wildtype mice Tpm3, Fgfrl1 (p = 0.004), Myl2, Lrtm1, Myh4, Myl3, Myh7 and Hephl1. Lrtm1 is closely related to Slit3, a protein associated with herniation of the central tendon of the diaphragm in mice. The Slit proteins are known to regulate axon branching and cell migration, and inhibition of Slit3 reduces cell motility and decreases the expression of Rac and Cdc42, two genes that are essential for myoblast fusion. Further studies to determine if Lrtm1 has a similar function to Slit3 and if reduced Fgfrl1 expression can cause diaphragm hypoplasia through a mechanism involving decreased myoblast motility and/or myoblast fusion, seem indicated. PMID- 20024585 TI - Nitrogen fluxes from treefrogs to tank epiphytic bromeliads: an isotopic and physiological approach. AB - Diverse invertebrate and vertebrate species live in association with plants of the large Neotropical family Bromeliaceae. Although previous studies have assumed that debris of associated organisms improves plant nutrition, so far little evidence supports this assumption. In this study we used isotopic ((15)N) and physiological methods to investigate if the treefrog Scinax hayii, which uses the tank epiphytic bromeliad Vriesea bituminosa as a diurnal shelter, contributes to host plant nutrition. In the field, bromeliads with frogs had higher stable N isotopic composition (delta(15)N) values than those without frogs. Similar results were obtained from a controlled greenhouse experiment. Linear mixing models showed that frog feces and dead termites used to simulate insects that eventually fall inside the bromeliad tank contributed, respectively, 27.7% (+ or 0.07 SE) and 49.6% (+ or - 0.50 SE) of the total N of V. bituminosa. Net photosynthetic rate was higher in plants that received feces and termites than in controls; however, this effect was only detected in the rainy, but not in the dry season. These results demonstrate for the first time that vertebrates contribute to bromeliad nutrition, and that this benefit is seasonally restricted. Since amphibian-bromeliad associations occur in diverse habitats in South and Central America, this mechanism for deriving nutrients may be important in bromeliad systems throughout the Neotropics. PMID- 20024586 TI - Severe respiratory adverse events associated with rituximab infusion. PMID- 20024587 TI - Cutaneous warts in children before and after renal transplantation. AB - Cutaneous warts occur in 3.9-4.9% of children in the UK. The incidence is increased in organ transplant recipients and may be increased in patients with chronic kidney disease (CKD), since uraemia reduces the immune system's function. We surveyed the records from our CKD and renal transplant clinic to ensure patients with warts were identified and appropriately treated. Data were collected by questionnaire. The presence of warts, location, treatment, levels of pain and emotional upset were recorded. Nine of 49 (18.4%) pre-transplantation patients (33 male, median age 12.1 years) were currently suffering from warts compared with 17 of 60 (28.3%) post-transplantation patients (34 male, median age 13.9 years). A further 14 pre-transplantation and 16 post-transplantation patients had previously suffered from warts which had resolved. Forty-one patients had sought treatment for warts, mainly from primary care. Five patients, all having received transplants, were seen by a dermatologist. Self-rated levels of pain and emotional upset were generally low, apart from those of four adolescent patients who expressed significant emotional upset. We concluded that cutaneous warts are more common among CKD patients. Appropriate information and treatment are required before and after transplantation. The majority of warts can be treated in primary care, but selected patients with extensive warts that cause distress need early referral for dermatology opinion. PMID- 20024588 TI - Spleen-preserving distal pancreatectomy with conservation of the splenic artery and vein: techniques and its significance. AB - BACKGROUND: Preservation of the spleen in distal pancreatectomy has recently attracted considerable attention. Since our first trial and success with spleen preserving distal pancreatectomy with conservation of the splenic artery and vein for tumors of the pancreas and chronic pancreatitis, this procedure (Kimura's procedure) has been performed very frequently. METHODS: The techniques for spleen preserving distal pancreatectomy (SpDP) with conservation of the splenic artery and vein are clarified. The splenic vein is identified behind the pancreas and within the thin connective tissue membrane (fusion fascia of Toldt). The connective tissue membrane is cut longitudinally above the splenic vein. It is important to remove the splenic vein from the pancreas by working from the body of the pancreas toward the spleen (median approach), because it is very difficult to remove it in the other direction. The pancreas is removed from the splenic artery by proceeding from the spleen toward the head of the pancreas. RESULTS: Preservation of the spleen offers various advantages. The maximum platelet levels in blood serum are significantly lower in postoperative patients with splenic preservation than in those with splenectomy. The platelet count was maximal on postoperative day 10 in the 16 patients with SpDP and the count was maximal on postoperative day 13 in the 16 patients with distal pancreatectomy with splenectomy (DPS), and there was a smaller increase in the patients with SpDP than in the patients with DPS. Postoperative bleeding from an ablated splenic artery and vein in SpDP has not been encountered. Either DPS or spleen preservation without preservation of the splenic artery and vein may reduce the blood supply to the residual proximal stomach after distal gastrectomy, which is different from the findings in the Kimura procedure. CONCLUSION: In SpDP, a very slight elevation of the platelet count in serum may help to prevent infarction of the lungs and brain compared to DPS. Another advantage of SpDP performed according to our procedure is that the blood supply to the proximal stomach is conserved in patients with SpDP who undergo distal gastrectomy with resection of the left gastric artery. Benign lesions, as well as low-grade malignancy of the body and tail of the pancreas, may be indications for this procedure. Surgeons should know the techniques and significance of SpDP with conservation of the splenic artery and vein, which is a very safe and reliable method. PMID- 20024589 TI - Neuraminidase-deficient Sendai virus HN mutants provide protection from homologous superinfection. AB - Binding of hemagglutinin-neuraminidase proteins (HN) to sialylated receptors initiates the infection process of several paramyxoviruses, whereas later in the viral life cycle, the neuramindase (NA) activity of newly synthesized HN destroys all receptors. Prior to NA action, expressed HN has to bind the receptor. To evaluate this HN-receptor complex with respect to receptor inactivation, three temperature-sensitive Sendai virus HN mutants carrying amino acid exchanges at positions 262, 264 and/or 461 were created that uncoupled NA activity from receptor binding at 39 degrees Celsius. Interestingly, at elevated temperature, when there is no detectable neuramindase activity, all infected cells are protected against homologous superinfection. Mutated HN protein on the cell surface is mainly bound to sialylated cell-surface components but can be released by treatment with NA. Thus, continuous binding to HN already inactivates the receptors quantitatively. Furthermore, mutant HN bound to receptors is prevented from being incorporated into virus particles in the absence of NA. It is shown here for the first time that during paramyxoviral infection, quantitative receptor inactivation already occurs due to binding of receptors to expressed HN protein without involvement of NA and is independent of NA activity of viral progeny. NA subsequently functions in the release of HN from the complex, coupled with desialysation of receptors. These findings could have implications for further antiviral drug development. PMID- 20024590 TI - Effects of the bisphosphonate risedronate on osteopenia in OASIS-deficient mice. AB - Endoplasmic reticulum (ER) stress has been reported to be linked to various diseases such as diabetes, neurodegenerative diseases, and osteogenesis imperfecta (OI). Old astrocyte specifically induced substance (OASIS), a novel type of ER stress transducer, is a basic leucine zipper transcription factor belonging to the CREB/ATF family and is markedly expressed in osteoblasts. Recently, we demonstrated that OASIS activates the transcription of the gene for type I collagen, Col1a1, and contributes to the secretion of bone matrix proteins in osteoblasts. OASIS-/- mice exhibit severe osteopenia involving a decrease in type I collagen in the bone matrix and a dysfunction of osteoblasts, which show abnormal expansion of the rough ER. These phenotypic features of osteopenia are similar to those observed in OI type I. In this study, we investigated whether administration of the third-generation bisphosphonate risedronate (RIS) is effective for treating osteopenia in OASIS-/- mice. Histological and histomorphometric analyses revealed that the trabecular bones increased dramatically in OASIS-/- mice treated with RIS, owing to the inhibition of bone resorption. Intriguingly, the abnormal expansion of the rough ER in OASIS-/- osteoblasts was improved by the treatment with RIS. Taken together, we conclude that OASIS-/- mice will be useful as new model mice for evaluating the medicinal effects of osteopenia treatments and developing new drugs for the osteopenia associated with diseases such as OI and osteoporosis. PMID- 20024591 TI - Hop rho iso-alpha acids, berberine, vitamin D3 and vitamin K1 favorably impact biomarkers of bone turnover in postmenopausal women in a 14-week trial. AB - Osteoporosis is a major health issue facing postmenopausal women. Increased production of pro-inflammatory cytokines resulting from declining estrogen leads to increased bone resorption. Nutrition can have a positive impact on osteoporosis prevention and amelioration. The objective of this study was to investigate the impact of targeted phytochemicals and nutrients essential for bone health on bone turnover markers in healthy postmenopausal women. In this 14 week, single-blinded, 2-arm placebo-controlled pilot study, all women were instructed to consume a modified Mediterranean-style low-glycemic-load diet and to engage in limited aerobic exercise; 17 randomized to the placebo and 16 to the treatment arm (receiving 200 mg hop rho iso-alpha acids, 100 mg berberine sulfate trihydrate, 500 IU vitamin D(3) and 500 microg vitamin K(1), twice daily). Thirty two women completed the study. Baseline nutrient intake did not differ between arms. At 14 weeks, the treatment arm exhibited an estimated 31% mean reduction (P = 0.02) in serum osteocalcin (a marker of bone turnover), whereas the placebo arm exhibited a 19% increase (P = 0.03) compared to baseline. Serum 25-hydroxyvitamin D (25(OH)D) increased by 13% (P = 0.24) in the treatment arm and decreased by 25% (P < 0.01) in the placebo arm. The between-arm differences for OC and 25(OH)D were statistically significant. Serum IGF-I was increased in both arms, but the increase was more significant in the treatment arm at 14 weeks (P < 0.01). Treatment with hop rho iso-alpha acids, berberine sulfate trihydrate, vitamin D(3) and vitamin K(1) produced a more favorable bone biomarker profile that supports a healthy bone metabolism. PMID- 20024593 TI - A retrospective overview of treatment choice and outcome in 126 cases with arrested eruption of mandibular second molars. AB - The purpose of the present retrospective study was to analyze treatment choice and outcome in patients with retention/impaction of the mandibular second molar. Radiographic material, from three large clinics, from 106 patients (60 males and 46 females) with 126 retained/impacted permanent mandibular second molars treated during the years 1985-2005 was evaluated for treatment choice and treatment outcome. Follow-up questionnaires were sent to dentists in cases where treatment outcome could not be determined from the radiographic material. Clinical evaluation was not possible as the patients were no longer associated with the clinic where they were treated. The cases were categorized into six groups: (A) no treatment; (B) orthodontic treatment; (C) surgical exposure of the second molar; (D) removal of the third molar; (E) removal of the second molar; and (F) other treatments. The various treatment choices performed during 1985-2005 showed acceptable results in 66 of the cases. In 23 cases, the results were unacceptable. In 37 cases, the radiographic material could not document the outcome nor was evaluation of the final outcome possible due to the patient's young age. Remarkable are the high percentages of unacceptable treatment outcome, 25.9% in group D (removal of third molar) and 23% in group E (removal of second molar). As the material was collected before new advanced methods of surgical uprighting and new methods of orthodontic uprighting had been introduced, these percentages are expectedly lower today. Even so, it is highly recommended to focus especially on these two groups in future studies on treatment outcome. PMID- 20024592 TI - Bisphosphonates: restrictions for vasculogenesis and angiogenesis: inhibition of cell function of endothelial progenitor cells and mature endothelial cells in vitro. AB - Bisphosphonate-associated osteonecrosis of the jaws (BP-ONJ) is one of the main side effects in patients treated with bisphosphonates for metastasis to the bone or osteoporosis. BP-ONJ usually occurs in patients treated with highly potent nitrogen-containing bisphosphonates. The exact mechanism of action and etiopathology is still unknown. In addition to inhibition of bone remodelling, an anti-angiogenetic effect has become the focus of research. The aim of these study was to investigate the effect of different bisphosphonates on human umbilicord vein endothelial cells (HUVEC) and endothelial progenitor cells (EPC), which play an important role in angiogenesis. Using varying concentrations, the impact of one non-nitrogen-containing bisphosphonate (clodronate) and three nitrogen containing bisphosphonates (ibandronate, pamidronate and zoledronate) on HUVEC and EPC was analysed. The biologic behaviour of HUVEC after incubation with different bisphosphonates was measured in a Boyden migration assay as well as in a 3D angiogenesis assay. The number of apoptotic cells was measured by Tunnel assay. To underline the importance of neoangiogenesis in the context of BP-ONJ, we measured the EPC number after incubation with different bisphosphonates in vitro. HUVEC and EPC were significantly influenced by bisphosphonates at different concentrations compared with the non-treated control groups. The nitrogen-containing bisphosphonates pamidronate and zoledronate had the greatest impact on the cells, whereas clodronate followed by ibandronate was less distinct on cell function. These results underline the hypothesis that inhibited angiogenesis induced by bisphosphonates might be of relevance in the development and maintenance of BP-ONJ. The increased impact by highly potent bisphosphonates on HUVEC and EPC may explain the high prevalence of BP-ONJ in patients undergoing this treatment. PMID- 20024594 TI - Chronic kidney disease management in an academic internal medicine clinic. AB - OBJECTIVE: We sought to evaluate the current state of chronic kidney disease (CKD) management in our academic internal medicine clinic. METHODS: A retrospective review was carried out involving all patients with laboratory evidence of CKD enrolled in our clinic. We evaluated the rate of CKD recognition as well as compliance with standard guidelines. We further subdivided our results based on physician training level, presence of diabetes, recognition of CKD, age, and race. RESULTS: Factors that significantly improved recognition and treatment of CKD in our study included presence of diabetes (p < 0.001), black race (p = 0.013), younger age (p = 0.004), and treatment by a resident physician (p = 0.009). Recognition of stage 3 CKD was associated with significant increases in urine protein analysis (p < 0.001) and nephrology consultation (p < 0.001). CONCLUSION: Chronic kidney disease remains under-recognized and undertreated despite well-publicized guidelines and widespread use of routine eGFR reporting. PMID- 20024595 TI - Lung and thoracic imaging. From the editor's desk. PMID- 20024597 TI - Receptive music therapy and salivary histamine secretion. AB - INTRODUCTION: Receptive music therapy (rMT) not only provides a good feeling but also a more effective healing process and mastery of stress. METHODS AND RESULTS: In a preliminary study it could be shown that American Doudouk-music (feel-good music) suppressed salivary histamine secretion in two groups (n = 4) of allergic and non-allergic young volunteers. Stress was induced by eating adverse food/allergenic food during music exposure. There was no response in the vein blood samples and no significant difference between the allergic and non-allergic groups. CONCLUSION: It can be concluded that saliva is an appropriate medium for histamine measurements during music exposure. PMID- 20024598 TI - Inflammation Research. Meeting report. PMID- 20024596 TI - From nature versus nurture, via nature and nurture, to gene x environment interaction in mental disorders. AB - It is now generally accepted that complex mental disorders are the results of interplay between genetic and environmental factors. This holds out the prospect that by studying G x E interplay we can explain individual variation in vulnerability and resilience to environmental hazards in the development of mental disorders. Furthermore studying G x E findings may give insights in neurobiological mechanisms of psychiatric disorder and so improve individualized treatment and potentially prevention. In this paper, we provide an overview of the state of field with regard to G x E in mental disorders. Strategies for G x E research are introduced. G x E findings from selected mental disorders with onset in childhood or adolescence are reviewed [such as depressive disorders, attention deficit/hyperactivity disorder (ADHD), obesity, schizophrenia and substance use disorders]. Early seminal studies provided evidence for G x E in the pathogenesis of depression implicating 5-HTTLPR, and conduct problems implicating MAOA. Since then G x E effects have been seen across a wide range of mental disorders (e.g., ADHD, anxiety, schizophrenia, substance abuse disorder) implicating a wide range of measured genes and measured environments (e.g., pre-, peri- and postnatal influences of both a physical and a social nature). To date few of these G x E effects have been sufficiently replicated. Indeed meta-analyses have raised doubts about the robustness of even the most well studied findings. In future we need larger, sufficiently powered studies that include a detailed and sophisticated characterization of both phenotype and the environmental risk. PMID- 20024599 TI - Improved production of polygalacturonate lyase by combining a pH and online methanol control strategy in a two-stage induction phase with a shift in the transition phase. AB - Polygalacturonate lyase is a kind of enzyme that is abundantly used in the textile industry for cotton scouring. Previously, we reconstructed the polygalacturonate lyase gene in Pichia pastoris for the expression of this enzyme. To enhance the production of polygalacturonate lyase (PGL), a combined strategy was formulated by combining online methanol control and two-stage pH control strategies. For the two-stage pH control strategy during the growth phase, the pH was controlled at 5.5, and in the induction phase different pH levels were investigated for the optimum enzyme production. During the online methanol control strategy, the different levels of methanol (v/v) were investigated for the best enzyme production at pH 5.5. These two strategies were combined together for enhanced PGL productivity, and the induction phase was divided into two stages in which methanol concentrations were maintained at different levels online. The transition phase was introduced during the induction phase instead of introducing it after the growth phase. The two-stage combination strategy was formulated on the bases of methanol consumption of cells, optimal pH, cell viability and the production of polygalacturonate lyase by P. pastoris. By using this strategy, the production was doubled compared with common conditions, and the highest polygalacturonate lyase activity reached 1,631 U/ml. This strategy proved to be very useful for the enhancement of polygalacturonate lyase production by achieving higher cell viability, alcohol oxidase activity and phosphate-related compounds of the cells during the induction phase. PMID- 20024600 TI - Microarray analysis of p-anisaldehyde-induced transcriptome of Saccharomyces cerevisiae. AB - p-Anisaldehyde (4-methoxybenzaldehyde), an extract from Pimpinella anisum L. seeds, is a potential novel preservative. To reveal the possible action mechanism of p-anisaldehyde against microorganisms, yeast-based commercial oligonucleotide microarrays were used to analyze the genome-wide transcriptional changes in response to p-anisaldehyde. Quantitative real-time RT-PCR was performed for selected genes to verify the microarray results. We interpreted our microarray data with the clustering tool, T-profiler. Analysis of microarray data revealed that p-anisaldehyde induced the expression of genes related to sulphur assimilation, aromatic aldehydes metabolism, and secondary metabolism, which demonstrated that the addition of p-anisaldehyde may influence the normal metabolism of aromatic aldehydes. This genome-wide transcriptomics approach revealed first insights into the response of Saccharomyces cerevisiae (S. cerevisiae) to p-anisaldehyde challenge. PMID- 20024602 TI - Robust structure-based resonance assignment for functional protein studies by NMR. AB - High-throughput functional protein NMR studies, like protein interactions or dynamics, require an automated approach for the assignment of the protein backbone. With the availability of a growing number of protein 3D structures, a new class of automated approaches, called structure-based assignment, has been developed quite recently. Structure-based approaches use primarily NMR input data that are not based on J-coupling and for which connections between residues are not limited by through bonds magnetization transfer efficiency. We present here a robust structure-based assignment approach using mainly H(N)-H(N) NOEs networks, as well as (1)H-(15) N residual dipolar couplings and chemical shifts. The NOEnet complete search algorithm is robust against assignment errors, even for sparse input data. Instead of a unique and partly erroneous assignment solution, an optimal assignment ensemble with an accuracy equal or near to 100% is given by NOEnet. We show that even low precision assignment ensembles give enough information for functional studies, like modeling of protein-complexes. Finally, the combination of NOEnet with a low number of ambiguous J-coupling sequential connectivities yields a high precision assignment ensemble. NOEnet will be available under: http://www.icsn.cnrs-gif.fr/download/nmr. PMID- 20024601 TI - Gli1 inhibition induces cell-cycle arrest and enhanced apoptosis in brain glioma cell lines. AB - The Hedgehog (HH)-Gli1 signaling pathway plays an important role in the patterning and development of the central nervous system during embryogenesis. Recent data have shown its possible involvement in a subset of human gliomas, and inhibition of the pathway resulted in tumor suppression in both in vitro and in vivo studies. The underlying mechanisms of tumor suppression, however, remain to be fully elucidated. Here, we investigated Gli1 expression in 65 surgically resected malignant glioma tissues and found the Ki-67 labeling index to be higher in Gli1-positive gliomas than in Gli1-negative gliomas. Depletion of Gli1 expression by small interfering RNA (siRNA) interference led to remarkably decreased cell proliferation and enhanced apoptosis in U87 glioma cell line. To explore the molecular mechanisms of the phenotypic changes, we performed real time quantitative RT-PCR analysis to monitor the changes of a series of genes which play critical roles in the regulation of cell cycle and apoptosis. The result showed that downregulation of G(1) cyclins, downregulation of Bcl-2, and upregulation of p21 were detected after Gli1 downregulation. Additionally, cyclopamine was used to inhibit the HH signaling activity as an indirect approach to decrease Gli1 expression, and we observed that cyclopamine exclusively inhibited cell growth in HH-pathway-active glioma cell lines. The cell phenotypic and molecular changes induced by cyclopamine were consistent with those caused by siGli1 interference. In conclusion, our findings support an important role of Gli1 in cell-cycle and apoptosis regulation in human brain gliomas; hence, it can serve as a potential target of new therapeutic strategies for these diseases. PMID- 20024603 TI - Can iron be teratogenic? AB - Several kinds of evidence indicate that elevated iron during the 3-8 week embryonic (organogenesis) period of human gestation may be teratogenic. (1) In the embryonic period, the natural maternal absorption of food iron is 30% below the estimated daily iron loss. (2) As compared with maternal serum, embryonic fetal coelomic fluid contains only one-fourth as much iron but nearly six times the quantity of the iron withholding protein, ferritin. (3) In the embryonic period, intraplacental oxygen pressure is 2-3 times lower than in the subsequent fetal growth period. (4) Iron is a strong inducer of emesis which peaks in the embryonic period. (5) In a murine gestation model, iron was neurotoxic at a sharp peak of 8-9 days. Thus it would be prudent, in human pregnancy, to delay any needed iron supplementation until the embryonic period has been completed. PMID- 20024605 TI - Diagnostic validity of hospital discharge codes in the ascertainment of amyotrophic lateral sclerosis. PMID- 20024604 TI - Upward trends in symptom reporting in the UK Armed Forces. AB - Several reports have shown increases in the prevalence of non-specific symptoms in the general population. Research in the military tends to focus on comparisons between deployed and non-deployed personnel and does not examine trends over time. 4,257 and 4,295 male participants of the Gulf war and Iraq war studies not deployed to either of these wars were randomly sampled and surveyed in 1997/1998 and 2004/2006 in two independent cross-sectional studies. Information was collected on 50 symptoms and the General Health Questionnaire (GHQ-12). Factor analysis was performed to identify an underlying pattern of symptom dimensions, and multivariate regressions were carried out to examine changes in symptom dimensions between the two surveys and the possible role of psychological morbidity. Factor analysis identified a robust pattern of eight symptom dimensions. An increase in the prevalence of symptoms was evident across all symptom dimensions. Adjustment for demographic and service characteristics revealed increases in the odds of scoring highly on symptom dimensions, varying from odds ratios 1.57, 95% CI 1.36-1.81 (cardio-respiratory dimension) to 2.24, 95% CI 1.93-2.60 (fatigue dimension). Unexpectedly, increases were even greater when adjusting for psychological morbidity. There is clear evidence of an increase in the reporting of non-specific symptoms over a 7 year period in the UK Armed Forces. It suggests that the threshold for reporting symptoms has decreased and cannot be explained by psychological distress. The possible implication of this trend for medical practice in the wider population deserves close scrutiny. PMID- 20024606 TI - Evidence for cross-talk between atrial natriuretic peptide and nitric oxide receptors. AB - Guanylyl cyclases (GCs), a ubiquitous family of enzymes that metabolize GTP to cyclic GMP (cGMP), are traditionally divided into membrane-bound forms (GC-A-G) that are activated by peptides and cytosolic forms that are activated by nitric oxide (NO) and carbon monoxide. However, recent data has shown that NO activated GC's (NOGC) also may be associated with membranes. In the present study, interactions of guanylyl cyclase A (GC-A), a caveolae-associated, membrane-bound, homodimer activated by atrial natriuretic peptide (ANP), with NOGC, a heme containing heterodimer (alpha/beta) beta1 isoform of the beta subunit of NOGC (NOGCbeta1) was specifically focused. NOGCbeta1 co-localized with GC-A and caveolin on the membrane in human kidney (HK-2) cells. Interaction of GC-A with NOGCbeta1 was found using immunoprecipitations. In a second set of experiments, the possibility that NOGCbeta1 regulates signaling by GC-A in HK-2 cells was explored. ANP-stimulated membrane guanylyl cyclase activity (0.05 +/- 0.006 pmol/mg protein/5 min; P < 0.01) and intra cellular GMP (18.1 +/- 3.4 vs. 1.2 +/- 0.5 pmol/mg protein; P < 0.01) were reduced in cells in which NOGCbeta1 abundance was reduced using specific siRNA to NOGCbeta1. On the other hand, ANP-stimulated cGMP formation was increased in cells transiently transfected with NOGCbeta1 (530.2 +/- 141.4 vs. 26.1 +/- 13.6 pmol/mg protein; P < 0.01). siRNA to NOGCbeta1 attenuated inhibition of basolateral Na/K ATPase activity by ANP (192 +/- 22 vs. 92 +/- 9 nmol phosphate/mg protein/min; P < 0.05). In summary, the results show that NOGCbeta1 and GC-A interact and that NOGCbeta1 regulates ANP signaling in HK 2 cells. The results raise the novel possibility of cross-talk between NOGC and GC-A signaling pathways in membrane caveolae. PMID- 20024607 TI - Up-regulated alpha-actin expression is associated with cell adhesion ability in 3 D cultured myocytes subjected to mechanical stimulation. AB - This study was aimed to investigate the alteration of alpha-actin in three dimensionally (3-D) cultured myocytes under cyclic tensile stress loading. Myocytes were collected from neonatal SD rat's lateral pterygoid muscle for primary cell culture. The third-passage cells were implanted and 3-D cultured in poly lactic-co-glycolic acid (PLGA) scaffold, and then subjected to cyclic tensile stress (0.5 Hz, 2,000 microstrain) for 0, 2, 4, 8, 12, and 24 h through a four-point bending strain system. The alpha-actin mRNA was investigated by semi quantitative RT-PCR. The alpha-actin protein expression was examined by immunofluorescent cytochemistry, laser confocal scanning microscopy (LCSM), and image analysis technology. The dynamic adhesion of myocytes to PLGA scaffolds was investigated by fluorescence microscope and the viability of the myocytes was measured by MTT assay. After mechanical loading, the alpha-actin mRNA increased at 2 h and then declined. The alpha-actin protein expression kept increased until peaked at 12 h, but declined at 24 h. The time course changing of alpha-actin protein expression parallelled with that of cell adhesion ability. It is concluded that alpha-actin expression is probably associated with cell adhesion ability in myocytes subjected to mechanical stimulation. PMID- 20024608 TI - No increase in criminal convictions in Hans Asperger's original cohort. AB - Hans Asperger originally used the term "autistic psychopathy" to describe his patients on the autism spectrum, leading to a possible confusion with psychopathic disorder and delinquent behaviour. We conducted a penal register search for 177 former patients of Asperger's clinic with a childhood diagnosis of "autistic psychopathy" or features of the disorder in Austria. The mean percentage of registered convictions was similar to that in the general male population of Austria over the studied time period. A qualitative assessment of offence types in Asperger's former patients suggests that the nature of offences does not differ from that in the general population. In this original cohort of Asperger's patients, convictions were no more common than in the general male population. PMID- 20024609 TI - Molecular characterization and phylogeny of two begomoviruses infecting Malvastrum americanum in Jamaica: evidence of the contribution of inter-species recombination to the evolution of malvaceous weed-associated begomoviruses from the northern Caribbean. AB - Two distinct full-length begomovirus DNA-A components and a DNA-B component were PCR amplified, cloned and sequenced from Jamaican Malvastrum americanum plants exhibiting yellow mosaic symptoms. Whereas one of the DNA-A components is from a potentially new species that we have tentatively named Malvastrum yellow mosaic Helshire virus (MaYMHV), the other DNA-A and the DNA-B form a cognate pair and represent a new virus species tentatively named Malvastrum yellow mosaic Jamaica virus (MaYMJV). The MaYMJV genome components together infected M. americanum and produced yellow mosaic symptoms similar to those seen in naturally infected plants. Both the MaYMJV and MaYMHV DNA-A components are typical of those of bipartite begomoviruses from the Western Hemisphere. The DNA-As of MaYMJV and MaYMHV are most closely related to each other (sharing 84% sequence identity) and cluster phylogenetically with begomoviruses found infecting malvaceous weeds in Cuba and Florida. The DNA-B component of MaYMJV is most similar to that of Sida golden mosaic virus-[USA:Florida] (SiGMV-[US:Flo]) and Sida golden mosaic Costa Rica virus-[Costa Rica] (SiGMCRV-[CR]). As with many other geminivirus species, the genomes of MaYMJV and MaYMHV bear traces of inter-species recombination. PMID- 20024610 TI - Racial differences in the incidence of breast cancer subtypes defined by combined histologic grade and hormone receptor status. AB - Breast cancer encompasses several distinct clinical entities of very different characteristics and behaviors, a fact which likely contributes to the higher breast cancer mortality in African-Americans (AA) despite the higher incidence in European-Americans (EA). We are interested in how incidence variability in cancer subtypes defined by combined estrogen receptor (ER) and grade contributes to racial mortality disparities. As an initial step, we compared age-specific and age-adjusted incidence rates for each ER/Grade subtype in South Carolina (SC-a southern state) with Ohio (a northern mid-western state), using state registry data for 1996-2004. Each ER/Grade subtype had a distinct incidence pattern and rate, with three striking racial/geographic differences. First, the racial incidence disparity in ER negative (ER-) cancers was mostly within the ER-/G3 subtype, of which AAs had ~65% higher incidence than did EAs; ER-/G2 was much less common, but of significantly higher incidence in AAs. Second, the racial disparity in ER positive (ER+) cancers was in the ER+/lower-grade cancers, with a marked EA excess in both states. Third, AA incidence of the ER+/lower-grade subtypes was ~26% higher in Ohio than in SC. The other subtypes (ER-/G1 and ER+/G3) varied minimally by race and state, and the latter showed a strong association with age. Age adjustment halved the racial difference in mean age at diagnosis to about 2 years younger in AAs, compared to 4 years younger in case comparisons. Use of age-adjusted and age-specific rates of breast cancer subtypes may improve understanding of racial incidence and mortality disparities over time and geography. This approach also may aid in estimating the race-specific incidence rates of triple-negative breast cancer. PMID- 20024611 TI - Erratum to: Glutamate regulation of calcium and IP(3) oscillating and pulsating dynamics in astrocytes. AB - [This corrects the article DOI: 10.1007/s10867-009-9155-y.]. PMID- 20024612 TI - Amplification of fibroblast growth factor receptor-1 in breast cancer and the effects of brivanib alaninate. AB - Fibroblast growth factor receptor-1 (FGFR-1) is amplified in 10% of human breast cancers. The goal of this study was to test the correlation between FGFR-1 amplification and expression and sensitivity to brivanib, an FGFR-1 small molecule inhibitor, in breast cancer cell lines in vitro. Using CGH array and gene expression profiling, FGFR-1 DNA copy number, mRNA, and protein expression were measured in 21 cell lines and correlated with growth inhibition by brivanib. We examined FGFR-1 autophosphorylation and kinase activity, as well as phosphorylation of downstream signaling molecules in response to bFGF and brivanib exposure. CAMA, MDA-MB-361, and HCC38 cells had FGFR-1 amplification and protein overexpression. Brivanib GI(50) values were significantly lower in the gene amplified (15.17 MUM, n = 3) compared to normal copy number (69.09 MUM, n = 11) or FGFR-1 deleted (76.14 MUM, n = 7) cells (P = 0.0107). Among nonamplified cells, there was no correlation between FGFR-1 mRNA or protein expression levels and brivanib sensitivity. Two of three FGFR-1 amplified cells were sensitive to bFGF-induced growth stimulation, which was blocked by brivanib. In cells with amplified FGFR-1, brivanib decreased receptor autophosphorylation, inhibited bFGF induced tyrosine kinase activity, and reduced phosphorylation of ERK and AKT. Breast cancer cell lines with FGFR-1 gene amplification and protein overexpression are more sensitive to growth inhibition by brivanib than nonamplified cells. These findings suggest that FGFR-1 amplification or protein overexpression in breast cancers may be an indicator for brivanib treatment, where it may have direct anti-proliferative effects in addition to its' anti angiogenic effects. PMID- 20024613 TI - Recommendations for research priorities in breast cancer by the coalition of cancer cooperative groups scientific leadership council: imaging and local therapy. AB - Imaging and local therapy are important modalities for detection and management of localized breast cancer. Improvements in screening and local therapy have contributed to reduced breast cancer-associated morbidity and mortality. The Coalition of Cancer Cooperative Groups (CCCG) convened the Scientific Leadership Council (SLC) in breast cancer, an expert panel, to identify priorities for future research and current trials with greatest practice-changing potential. Panelists formed a consensus on research priorities for breast imaging and locoregional therapy, and also identified six trials judged to be of high priority. Current high priority trials included trials determining: (1) the role of accelerated partial breast versus whole-breast radiation (B39), (2) the feasibility, safety, and local and systemic control of small localized breast cancers treated with tumor ablation (Z1072), (3) the role of removal of the primary cancer in selected patients with metastatic disease (E2108), and (4) the clinical and biological effects of pre-operative anti-HER2-directed and ER directed therapies in localized or locally advanced breast cancer (B41, Z1031, Z1041). Ongoing and future trials will further refine optimal locoregional management, and additional research is required to develop improved screening methods and identify high risk populations most likely to benefit from targeted screening. PMID- 20024614 TI - Concentrations, distributions and critical level exceedance assessment of SO2, NO2 and O3 in South Africa. AB - South Africa has been identified as a source of industrial pollution that is significant at a global scale. This study was designed to provide quantitative information, by direct measurement, across northeastern South Africa, which includes the highly industrialised Mpumalanga Highveld. The specific aim of the study was to evaluate whether or not acidic atmospheric pollution poses a threat to soils, plants and water bodies of South Africa. To address this aim, a network of 37 passive sampling sites was established to measure monthly mean concentrations of near-surface SO(2), NO(2) and ozone. The area covered extended over the northern and eastern interior of South Africa while avoiding sources of local emissions such as towns, mines and highways. The field campaign was conducted between August 2005 and September 2007. Spatial distributions and temporal trends for these pollutant gases were assessed. Critical levels analysis comparisons were made against applicable air quality standards, guidelines and limits to evaluate the potential for adverse atmospheric pollution impacts on regional environments. The assessment indicates that only in the central source area of the South African industrial Highveld are some levels exceeded. In remote areas, including the sensitive forested regions of the Drakensberg escarpment, pollutant concentrations are below the critical thresholds for environmental damage. PMID- 20024615 TI - Riparian vegetation assessment of Cauvery River Basin of South India. AB - The Cauvery river basin of South India has a large phyto-floristic wealth, rightfully enough to constitute a separate phyto-geographic unit. Increase in the anthropogenic pressures within the river basin and surrounding landscapes have persistently stressed the riparian ecosystem structure adversely, besides altering its composition. The objective of this study was to examine the present status of riparian vegetation along the Cauvery river basin, in response to anthropogenic pressures. For vegetation analysis, the riparian forest coming in the middle stretch of Cauvery river was categorized into two zones, viz., forest zone covering ~54 km stretch and agroecosystem zone covering ~80 km stretch. In each zone, tree species were quantified using transects at 8-km interval. Overall tree species accounting for both forest and agroecosystem were recorded and compared. The results indicate that the mean density and basal area of trees per plot were higher in the forest zone than agroecosystem zone. The Shannon-Weiner diversity of forest zone is 4.6, which is higher than agroecosystem. In addition, species composition indicated a relatively low or poor similarity between the two zones. The vegetation density and site disturbance scores for all the study sites reveals that sand mining and grazing areas have exerted negative impact on riparian forest. The results of the present study clearly brought out the need for preparing and implementing site-specific conservation plans for riparian ecosystem. PMID- 20024616 TI - Influence of phosphorus on copper sensitivity of fluvial periphyton: the role of chemical, physiological and community-related factors. AB - The influence of eutrophication of fluvial ecosystems (caused by increased phosphorus concentrations) on periphyton Cu sensitivity is explored from a multi scale perspective, going from the field to the laboratory. The study design included three tiers: a field study including the characterization of land use and the ecological state of the corresponding river sections in the Fluvia River watershed, an experimental investigation performed with natural periphyton from the previously studied stream sites in indoor channels, and finally a culture study in the laboratory. Results showed that differences in copper sensitivity of natural periphyton communities followed the gradient of nutrient concentration found in the field. Results from the culture experiments demonstrated that both, P-conditions during growth and P-content in the media are important factors modulating the toxicological response of algae to Cu. The observations from this study indicate that the ecological effects of metal pollution in rivers might be obscured by eutrophication. PMID- 20024619 TI - Protective effects of HFE7A, mouse anti-human/mouse Fas monoclonal antibody against acute and lethal hepatic injury induced by Jo2. AB - HFE7A is a mouse anti-human/mouse Fas monoclonal antibody which, protects mice from fulminant hepatitis induced by Jo2. Herein, we report on the mechanism of the protective effect of HFE7A against Jo2-induced acute and lethal hepatic injury. HFE7A reduced the serum aminotransferase level which was elevated after Jo2 injection. HFE7A also inhibited caspase activation and mitochondrial depolarization in hepatocytes derived from apoptosis induced by Jo2 injection. The protective effect of HFE7A against Jo2-induced apoptosis in mouse hepatocytes was reproducible in vitro. The cell death and caspase activation in isolated mouse hepatocytes were induced by incubating these cells with Jo2 in vitro, and HFE7A inhibited the cell death and caspase activation in mouse hepatocytes in a dose-dependent manner. The affinity of HFE7A to mouse Fas was lower than that of Jo2. The binding of Jo2 to neither recombinant mouse Fas nor mouse hepatocytes was inhibited by an excessive amount of HFE7A. Interestingly, HFE7A bound to hepatocytes isolated from Fas knockout mice. From these results, it is suggested that HFE7A may exert a protective effect against Jo2-induced hepatitis not by competitively inhibiting the binding of Jo2 to Fas on hepatocytes, and that a distinct molecule other than Fas may possibly be involved in the protective effect of HFE7A against Jo2-induced hepatic injury. PMID- 20024617 TI - Growth and antioxidant response in Hydrocharis dubis (Bl.) Backer exposed to linear alkylbenzene sulfonate. AB - A two-week exposure experiment was designed to investigate the toxicity of linear alkylbenzene sulfonate (LAS) on the aquatic plant Hydrocharis dubis (Bl.) Backer, focusing on growth, photosynthetic pigments and the activities of antioxidant enzymes. No significant differences were observed in the growth parameters of H. dubis when H. dubis was exposed to lower LAS doses (10 mg l(-1)) resulted in significant decreases in all growth parameters of H. dubis. No significant effect on pigment contents was observed at up to 50 mg l(-1) LAS, beyond which pigment contents declined gradually. Malondialdehyde (MDA) content did not show obvious differences when H. dubis plants were exposed to 100 times) over manual-marking whilst achieving a highly satisfactory sensitivity (0.92) and positive-prediction value (0.89). PMID- 20024625 TI - End-of-life ethics and disability: differing perspectives on case-based teaching. AB - The way in which medical professionals engage in bioethical issues ultimately reflects the type of care such patients are likely to receive. It is therefore critical for doctors and other health care professionals to have a broad understanding of disability. Our purpose in this paper is to explore ways of teaching bioethical issues to first year medical students by integrating alternative approaches. Such approaches include (a) the use of the narrative format, (b) the inclusion of a disability perspective, and (c) the presentation and facilitation of classes by people with disabilities. We consider how these new kinds of presentations are evaluated by students, faculty, people with disabilities and professional ethicists. We hope new knowledge may provide health care professionals with a greater understanding of the perspectives of patients with disabilities, who are confronted by conflicting ethical values and frameworks for decision-making in their interaction with such professionals. PMID- 20024627 TI - Population-based normative values for the Western Ontario and McMaster (WOMAC) osteoarthritis index and the Australian/Canadian (AUSCAN) hand osteoarthritis index functional subscales. AB - AIM: To develop age- and gender-specific normative values for the physical function subscales of the WOMAC and AUSCAN Indices. METHODS: A scannable survey questionnaire capable of capturing WOMAC and AUSCAN Index and demographic information was developed, pre-tested, and distributed to a stratified random sample of 24,000 members of the Australian general public generated by the Australian Electoral Commission (AEC). RESULTS: Age- and gender-specific WOMAC and AUSCAN normative values were estimated based on approximately 5,500 subjects. Age-related differences were noted at the subscale level. In general, disability increased with age for all items and both Indices. CONCLUSIONS: Normative values provide opportunity for benchmarking the health status of individuals against their age- and gender-matched peers in the general population. These normative values provide unique opportunities, for using the WOMAC and AUSCAN Indices in benchmarking applications, in both clinical practice and research. PMID- 20024626 TI - Melatonin formation in mammals: in vivo perspectives. AB - Melatonin is a hormone secreted from the pineal gland specifically at night and contributes to a wide array of physiological functions in mammals. Melatonin is one of the most well understood output of the circadian clock located in the suprachiasmatic nucleus. Melatonin synthesis is controlled distally via the circadian clock located in the suprachiasmatic nucleus and proximally regulated by norepinephrine released in response to the circadian clock signals. To understand melatonin synthesis in vivo, we have performed microdialysis analysis of the pineal gland, which monitors melatonin as well as the precursor (serotonin) and intermediate (N-acetylserotonin) of melatonin synthesis in freely moving animals in realtime at high resolution. Our data revealed a number of novel features of melatonin production undetected using conventional techniques, which include (1) large inter-individual variations of melatonin onset timing; (2) circadian regulation of serotonin synthesis and secretion in the pineal gland; and (3) a revised view on the rate-limiting step of melatonin formation in vivo. This article will summarize the main findings from our laboratory regarding melatonin formation in mammals. PMID- 20024628 TI - Silencing of CT120 by antisense oligonucleotides could inhibit the lung cancer cells growth. AB - BACKGROUND AND AIMS: The CT120 gene had been proven to be a novel gene closely related to pulmonary carcinogenesis and cancer progression. Our aim was to explore the mechanism of growth suppression caused by silencing CT120. METHODS: CT120 was detected in lung cancer tissues and the cell line A549, and the cell clones for silencing CT120 were obtained. Then the target genes were detected and the downstream proteins from the silencing of CT120 were separated and identified. RESULTS: The expression of CT120 was higher in lung cancer tissues and A549 cells. Silencing of CT120 was shown to inhibit cell growth, reduce the expression of cyclin D1 and Cdk4, and increase the expression of p53 and caspase 3. The differential proteins were related to carcinogenesis, invasiveness, and metastasis. CONCLUSION: CT120 may play an important role in tumor progression, and the down-regulation of CT120 expression could be a new drug target candidate in the treatment of lung cancer. PMID- 20024629 TI - Microelements in stones, urine, and hair of stone formers: a new key to the puzzle of lithogenesis? AB - The role of trace elements in lithogenesis is still unclear. The aim of this study was to evaluate the levels of elements in urinary stones and in the urine and hair of stone formers to identify these elements that have synergic correlations in studied materials and may contribute to lithogenesis. A total of 219 consecutive patients with idiopathic upper urinary tract stones were prospectively enrolled in the study. Urine and hair samples were collected from all patients. The content of the stone was evaluated using atomic absorption spectrometry, spectrophotometry, and colorimetric methods. The analysis of 29 elements in stones and hair and 21 elements in urine was performed using inductively coupled plasma-atomic emission spectrometry. The strength of correlation was described with the value of Spearman's rank correlation coefficient. The positive correlation between concentration of sodium, potassium, magnesium, barium, vanadium, zinc, silicon, phosphorus, and iodine in phosphate stones was observed. Only a few incidental correlations between the composition of stones and the distribution of elements in urine and in hair were found. There were 109 positive two-element correlations between two materials. The most common were observed for vanadium, aluminum, lead, cobalt, and molybdenum. Two-element positive correlations for all samples were established only for three elements: vanadium, lead, and aluminum. Results indicate that analysis of particular elements in hair and urine cannot predict the composition of urinary stones. This study showed, for the first time, correlations between the levels of vanadium, lead, and aluminum in the stones, urine, and hair of stone formers. PMID- 20024630 TI - Clinical characteristics and outcomes of women with stage IV endometrial cancer. AB - Treatment strategies for patients with stage IV endometrial cancer (EC) remain controversial. Some studies have suggested that optimal cytoreduction improves survival. We retrospectively analyzed the clinical characteristics and outcomes of 41 women with stage IV EC. The results of preoperative cytologic evaluation and biopsy of the endometrium were reviewed by a single pathologist for patients in whom stage IV EC was diagnosed preoperatively. Of the 41 patients with stage IV EC (median age, 62 years), 31 had surgical stage IV disease and 10 had clinical stage IV disease. Twenty-eight patients were diagnosed of stage IV EC before surgery or without surgery. Progression-free survival and overall survival were 10.4 and 21.3 months, respectively. On univariate analysis, grade 1 or 2 endometrioid subtype, 0 or 1 sites of extraperitoneal metastasis, and hormonal therapy were associated with good outcomes. Multivariate analysis revealed that grade 1 or 2 endometrioid subtype (P=0.005, hazard ratio [HR] 0.23 [0.08-0.65]) and 0 or 1 sites of extraperitoneal metastasis (P=0.001, HR 0.24 [0.10-0.57]) were independent predictors of survival. Neither surgery as primary therapy nor optimal cytoreduction was significantly related to overall survival in either the 28 patients in whom stage IV was diagnosed preoperatively or in all 41 patients. In women with stage IV EC, histologic features and extent of disease are more important determinants of outcomes than any kind of treatment. The indication for surgery should be carefully considered in this subset of patients. PMID- 20024631 TI - Pathology of retinal hemorrhage in abusive head trauma. AB - Abusive head injury, characterized by repeated acceleration-deceleration forces, is associated with retinal hemorrhages as demonstrated in many clinical and postmortem studies. The theory that vitreoretinal traction is the major factor in the pathogenesis of retinal hemorrhages is presently the most widely accepted explanation based on different lines of research. Postmortem examination of the eye and orbital structures is essential for recognizing abusive head injury and also for identifying other possible medical conditions which can cause retinal hemorrhage. PMID- 20024632 TI - Letter to the editor: Use of a trochanteric flip osteotomy improves outcomes in Pipkin IV fractures. PMID- 20024633 TI - T cell density and location can influence the prognosis of ovarian cancer. AB - The aims of this study were to examine the significance of CD3+ cells in patients with epithelial ovarian cancer and to determine their influence on the disease in relation to their location within tumours. A 157-core tissue-microarray constructed from primary ovarian cancer patients treated at Nottingham-University Hospitals (2000-2007) was stained for the T-cell marker CD3. The number of CD3+ cells in direct contact with tumour cells was counted per tumour area. These were considered as "intra-tumoural T-cells (ITTC)". Cores were divided into CD3 'high' or 'low' density tumours. "Stromal T-cells (STC)" were assigned as 'positive' or 'negative'. The study population had a median follow-up time of 36-months (0-75). The number of ITTC counted in tumour cores ranged between 0 and 184/mm(2). 90 tumours-(57%) were found to be in the "low-density" rubric, while 56-(36%) were of a "high-density" T-cell population. STC were found in 118-cores-(75%)-compared to 22-cores-(14%)-negative cores. Higher number of ITTC correlated with lower grade-(p = 0.045), tumour-type-(p = 0.034), and longer-median-survival-times (57 versus 37-months for high-and low-ITTC densities, respectively, p = 0.038). This relationship was reversed when tumours were infiltrated by CD3+ cells in the stroma, predicting worse-survival (Log-rank-test, p = 0.028). Combining ITTC with STC produced an interesting pattern where the ITTC-low/STC + ve had the worst prognosis (p = 0.003). Infiltration of ovarian cancer by T-cells can influence its prognosis depending on the location of these cells (intra-tumoural-versus stromal). The former predicts improved survival, while the latter is probably contributing to tumour progression and, in turn, worse survival. PMID- 20024634 TI - Effects of nonexhaustive bouts of high-intensity intermittent swimming training on GLUT-4 expression in rat skeletal muscle. AB - We previously reported that 14 bouts of exhaustive high-intensity intermittent training [20 s periods of swimming while carrying a weight (14% of body weight), separated by pauses of 10 s] is the highest stimuli in terms of exercise training induced glucose transporter 4 (GLUT-4) expression in rat epitrochlearis (EPI) muscles. In the present study, we found that the GLUT-4 protein content in the skeletal muscle of male Sprague-Dawley rats (age 5 weeks old; body weight 90-110 g) that underwent intermittent exercise training of 3 and 14 bouts of 20 s swimming for 5 days was increased over age-matched sedentary control rats by 75 and 71%, respectively, 18 h after the last bout of exercise. These results suggest that GLUT-4 content in rat EPI muscle increases dramatically after very short (60 s) and nonexhaustive high-intensity intermittent exercise training. PMID- 20024635 TI - Biomarker and no end to it? PMID- 20024636 TI - Biomarkers in heart failure--better than history or echocardiography? AB - The management of heart failure patients (diagnosis, treatment monitoring, assessment of prognosis) crucially depends on the accuracy of the tools used for patient evaluation. While the diagnostic accuracy of anamnesis, clinical signs and symptoms is poor, "classic" tools like chest X-ray or electrocardiogram show reasonable specificity but poor sensitivity. Transthoracic echocardiography is well suited for the diagnosis of heart failure, but is relatively costly and depends on availability and the skills of the investigator. Over the last 2 decades, numerous biomarkers have emerged that potentially might aid in the complex decisionmaking processes for diagnosis, treatment and monitoring of heart failure. The ideal biomarker would possess a favorable diagnostic test profile and deliver pivotal information. In this respect, the currently best evidence base has been accumulated for the natriuretic peptide family. A large array of other heart failure biomarkers (i.e., markers of inflammation, extracellular matrix remodeling, and myocyte damage and stress) are still in earlier phases of testing. This short review will provide a balanced comparison of those "classic tools" and natriuretic peptides focusing on their diagnostic capacity in various clinical settings, factors influencing test values, cutoff values and their interpretation, and the capacity of these biomarkers to guide the diagnostic and therapeutic decision-making processes in patients with suspected and established heart failure. PMID- 20024637 TI - Biomarkers for chronic heart failure : diagnostic, prognostic, and therapeutic challenges. AB - Cardiac biomarkers are very important in diagnosis, risk stratification, and management of patients with heart failure. Although not meeting all criteria for an ideal biomarker, natriuretic peptides primarily have a diagnostic and prognostic role. Other routinely available and newly emerging biomarkers have a complementary role in patient management; thus multimarker strategy might be warranted in future. The quest for a single marker or a combination is ongoing and several established, widely available biomarkers might have been overlooked in the field of heart failure. The authors review some of those biomarkers and speculate on the possible roles of combining two or more of them. PMID- 20024638 TI - Growth differentiation factor-15: a new biomarker in cardiovascular disease. AB - Growth differentiation factor-(GDF-)15 is a stress-responsive cytokine that is emerging as a biomarker of cardiac and vascular dysfunction and disease. Elevated circulating levels of GDF-15 identify high-risk individuals across the cardiovascular continuum, from stable coronary artery disease to acute coronary syndrome and heart failure. The association of GDF-15 with outcome in these conditions is independent of clinical risk factors and established biomarkers, including NT-proBNP (N-terminal pro-B-type natriuretic peptide) and troponin. The prognostic information provided by GDF-15 in cardiovascular disease may inform patient management, e.g., by identifying patients with non-ST segment elevation acute coronary syndrome who benefit from an invasive strategy, or by monitoring treatment response in heart failure. Future studies need to evaluate prospectively whether GDF-15, alone or as part of a multimarker strategy, can improve contemporary risk prediction algorithms and support therapeutic management of patients with cardiovascular disease. PMID- 20024639 TI - Troponins and high-sensitivity troponins as markers of necrosis in CAD and heart failure. AB - For years, cardiac troponins (cTn) have been regarded as the preferred biomarkers for the diagnosis of myocardial infarction and for the risk stratification of patients with acute coronary syndromes, as well as for the selection of patients who need an early invasive strategy, and for the guidance of adjunctive pharmacological therapy. In addition, measurement of cTn has been found useful for detection of myocardial necrosis in conditions unrelated to myocardial ischemia including acute pulmonary embolism, myocarditis, heart failure, sepsis, and end-stage renal disease. In these conditions, an unfavorable prognosis is unequivocally associated with detectable concentrations of cTn.A major limitation of most currently available cTn assays is the lack of adequate precision, i.e., to measure cTn concentrations at the 99th percentile value with a coefficient of variation < 10%. As a consequence, many manufacturers have developed more sensitive cTn assays that now comply with precision criteria required by the Joint European Society of Cardiology/ American College of Cardiology/American Heart Association/World Heart Federation Task Force for the Redefinition of Acute Myocardial Infarction.Using assays with higher analytic sensitivity more patients will be seen in clinical practice with the high-sensitivity cardiac troponin T (TnThs) above the 99th percentile discriminator. The causes of these elevations may be due to acute, subacute and chronic cardiac disease such as heart failure or cardiomyopathies. PMID- 20024640 TI - CRP in cardiovascular disease. AB - In primary prevention, traditional risk factors are a useful first step in determining who is at cardiovascular risk, however, it has been noted that a considerable number of those at risk cannot be identified on the basis of traditional risk factors alone. Among blood biomarkers, C-reactive protein (CRP), measured by high-sensitivity assays (hsCRP), has received widespread interest and a large database has been accumulated on its potential role as a predictor of cardiovascular risk, although observed associations between circulating CRP and coronary heart disease (CHD) are unlikely to be causal, as recently indicated by various Mendelian randomization studies. In a meta-analysis of 22 prospective studies, the multivariable adjusted, combined odds ratio for CRP to predict CHD, comparing extreme tertiles, was 1.58 (95% confidence interval, 1.48-1.68). Several recent studies showed a significant contribution of CRP to coronary risk prediction independent of the Framingham Risk Score, with better discrimination, calibration and improved, albeit modest reclassification of subjects at risk. To test the hypothesis whether or not subjects with normal low-density lipoprotein cholesterol but elevated CRP represent a population at increased risk that might benefit from statin treatment, the JUPITER trial randomized 17,802 apparently healthy persons to either 20 mg rosuvastatin daily or placebo. Rosuvastatin significantly reduced the incidence of major cardiovascular events. The rates of the primary endpoint (composite of nonfatal myocardial infarction, nonfatal stroke, hospitalization for unstable angina, revascularization, and confirmed death from cardiovascular causes) were 0.77 and 1.36 per 100 person-years of follow-up in the rosuvastatin and placebo groups, respectively. Relative risk reduction was 44%. CRP may have more accurately selected high-risk subjects due to its association with very many risk factors, thus representing an integrative marker of the total inflammatory burden of an individual. JUPITER has revitalized the discussion on CRP in clinical practice and will make it more difficult in the future to neglect the evidence built around CRP and cardiovascular risk. PMID- 20024641 TI - Biomarkers in inflammatory and noninflammatory cardiomyopathy. AB - Acute myocarditis (AMC) and its sequela, dilated cardiomyopathy (DCM), are most often caused by cardiotropic viral infections in the Western world. Inflammatory cardiomyopathy (DCMi) is a specific cardiomyopathy entity of DCM, being defined by the proof of intramyocardial inflammation and/or viral infection in endomyocardial biopsies (EMBs). Diagnostic procedures of EMBs are indispensable for the etiopathogenic differentiation of the disease. Experienced cardiology centers have reported low complication rates of EMB obtainment. The histological Dallas criteria are prone to substantial sampling error and interobserver variability, have no prognostic impact and, moreover, are not suitable to select AMC/DCMi patients who favorably respond to immunosuppression. Immunohistological detection of myocarditis and viral persistence have proven adverse prognostic impact in AMC and DCM patients, respectively. This contemporary diagnostic repertoire on EMBs is essential for the selection of DCMi patients who will likely benefit from immunomodulatory treatment, which has been addressed in randomized trials. During the past decade, cardiac magnetic resonance (CMR) has developed as a valuable noninvasive diagnostic approach for the detection and localization of intramyocardial inflammation, and CMR guidelines for AMC have been elaborated. Late gadolinium enhancement (LGE) has been associated with adverse prognostic outcome in DCM patients. CMR techniques, however, are not suitable to specifically detect myocardial viral infections. To date, no classic biomarker has been shown to differentiate between DCMi and noninflammatory cardiomyopathies. PMID- 20024642 TI - [Differentiation of malignant from nonmalignant, inflammatory pericardial effusions with biomarkers]. AB - Disorders of the pericardium are commonly associated with pericardial effusion. Its etiology comprises a broad spectrum of diseases including also malignancies. Pericardiocentesis, pericardioscopy and targeted epicardial biopsy with consecutive pericardial fluid and epicardial biopsy analysis by cytology, molecular biology and immunology establish the underlying etiology in the majority of cases. Of particular therapeutic and prognostic importance is the definite differentiation of malignant pericardial effusion from benign pericardial effusion. Biomarkers for cardiovascular diseases can be divided into biochemical, histological, immunologic, serologic and molecular markers as well as imaging biomarkers. Biomarkers have proven to be useful in the diagnosis, differential diagnosis and prognosis of ischemic heart disease and heart failure. With respect to pericardial disorders, a comprehensive approach combining clinical information, imaging biomarkers, biomarkers of pericardial effusion and analysis of epicardial biopsies often leads to the definite etiologic diagnosis of pericardial effusion. Computed tomography and magnetic resonance imaging allow further characterization of the effusion and, of note, also of the surrounding tissue, which is of particular interest in case of malignancies. Biomarkers of pericardial effusion include biochemical markers, autoantibodies, tumor markers, and cytokines. Analysis of pericardial fluid specific gravity, protein level and lactate dehydrogenase (LDH) separates transudates from exsudates. High adenosine deaminase levels (ADA) and low levels of carcinoembryonic antigen (CEA) in the pericardial effusion are observed in tuberculous pericarditis allowing the differentiation from malignant pericardial effusion. Additional markers, such as interferon and lysozyme, have also been suggested for the diagnosis of tuberculous pericarditis. Tumor markers in pericardial fluid have been used to diagnose malignant pericarditis. CEA levels are significantly higher in malignant than benign effusion. By a cutoff level of CEA > 5 ng/ml the diagnostic sensitivity and specificity are 75% and 100%, respectively, in the diagnosis of malignant pericardial effusion. Further analysis of cytokines and mediators, serologic, immunologic and inflammatory markers may help to understand the pathophysiology of the pericardial disease and provide useful diagnostic information. PMID- 20024643 TI - [BNP in HIV-infected patients]. AB - BACKGROUND AND PURPOSE: HIV-infected patients exhibit an increased rate of cardiac diseases, due to an elevated rate of cardiac risk factors and side effects of antiretroviral therapy. The aim of the present study was to analyze the impact of B-type natriuretic peptide (BNP) in this patient population. PATIENTS AND METHODS: The HIV-HEART (HIV infection and HEART disease) study is a prospective and multicenter cohort study. Outpatients with a known HIV infection were included over a 20-month recruiting period. BNP values were determined by a standardized laboratory test at study inclusion. RESULTS: 802 HIV-infected patients (male: 83.4%, mean age: 44.2 +/- 10.3 years) were included. The BNP concentrations (median 11.3 pg/ml; BNP < or = 100 pg/ml: 96.2%) were associated with echocardiographic parameters and with the right ventricular diameter (p = 0.02) and the systolic pulmonary arterial pressure (p = 0.01). Patients with a BNP concentration of > 50 pg/ml had a significantly higher rate of heart failure (p < 0.001), cardiomyopathy (p < 0.001), and coronary artery disease (p < 0.001). CONCLUSION: The data demonstrate that BNP is suitable for the detection of cardiac disorders in HIV-infected subjects. Therefore, BNP could be an appropriate tool for a screening program for HIV-associated disorders in this patient population. PMID- 20024644 TI - Myocardial dissection due to temporary cardiac pacemaker. PMID- 20024645 TI - Transfemoral aortic valve implantation in a patient with prior mechanical mitral valve replacement. AB - Transfemoral aortic valve implantation has emerged as a promising alternative to surgical aortic valve replacement for high-risk patients with severe symptomatic aortic stenosis. In the setting of previous mechanical mitral valve replacement, the procedure represents a challenge due to the risk of interference and subsequent functional impairment of the mechanical prosthesis. The authors report a case of successful transfemoral implantation of a selfexpandable aortic bioprosthesis in a patient with a Bjork-Shiley tilting-disk valve in mitral position demonstrating that the implantation is also feasible in this setting but requires careful preinterventional evaluation. Prior balloon aortic valvuloplasty with thorough observation of the mitral prosthesis during balloon inflation may be a helpful tool for indicating feasibility of this approach. PMID- 20024647 TI - MR features of the levator ani muscle in the immediate postpartum following cesarean delivery. AB - INTRODUCTION AND HYPOTHESIS: We wanted to show that at the time of cesarean delivery, the active process of labor itself was sufficient to create early alterations of the pelvic floor musculature as detectable via magnetic resonance imaging (MRI). METHODS: Thirty primipara patients underwent pelvic MRI between the second and third day after cesarean delivery. Ten patients had a cesarean without labor while 20 patients underwent urgent cesarean delivery after the onset of labor. RESULTS: Patients undergoing active labor during cesarean had 2.7 times more abnormalities than the patients with cesareans without labor. The abnormalities the most frequently found were a hypersignal in the puborectalis (p = .004), a hypersignal in the iliococcygeus (p = .064) and a defect in the orientation of this same muscular bundle (p = .049). CONCLUSION: This preliminary study suggests that active labor during the time of a cesarean induces early lesions of the pelvic muscular floor. PMID- 20024648 TI - Preliminary findings with the Solyx single-incision sling system in female stress urinary incontinence. AB - INTRODUCTION AND HYPOTHESIS: The aim of this study was to test the safety and efficacy of the Solyx single-incision sling (SIS) in women with stress urinary incontinence (SUI). METHODS: A retrospective study of 63 women who had undergone implantation at three medical centers (December-March 2009) was conducted. RESULTS: Mean patient age was 51 years (range, 30-87 years); 37 (59%) patients had concomitant urogynecological procedures. At a mean follow-up of 6.5 months (range, 5-8 months), 95% of patients were dry on the basis of subjective and objective assessment. Two patients experienced transient urinary retention, which resolved spontaneously. No complications of the procedure were reported, including no bladder, bowel, vessel, or nerve perforations and no erosions or extrusions. No pain was reported that was attributed to the implant. CONCLUSIONS: The very early efficacy results indicate that the Solyx SIS system is an attractive treatment option for SUI. The minimal number of steps involved in the procedure will facilitate uptake of the technique. PMID- 20024649 TI - Evaluation of an hPXR reporter gene assay for the detection of aquatic emerging pollutants: screening of chemicals and application to water samples. AB - Many environmental endocrine-disrupting compounds act as ligands for nuclear receptors. Among these receptors, the human pregnane X receptor (hPXR) is well described as a xenobiotic sensor to various classes of chemicals, including pharmaceuticals, pesticides, and steroids. To assess the potential use of PXR as a sensor for aquatic emerging pollutants, we employed an in vitro reporter gene assay (HG5LN-hPXR cells) to screen a panel of environmental chemicals and to assess PXR-active chemicals in (waste) water samples. Of the 57 compounds tested, 37 were active in the bioassay and 10 were identified as new PXR agonists: triazin pesticides (promethryn, terbuthryn, terbutylazine), pharmaceuticals (fenofibrate, bezafibrate, clonazepam, medazepam) and non co-planar polychlorobiphenyls (PCBs; PCB101, 138, 180). Furthermore, we detected potent PXR activity in two types of water samples: passive polar organic compounds integrative sampler (POCIS) extracts from a river moderately impacted by agricultural and urban inputs and three effluents from sewage treatment works (STW). Fractionation of POCIS samples showed the highest PXR activity in the less polar fraction, while in the effluents, PXR activity was mainly associated with the dissolved water phase. Chemical analyses quantified several PXR-active substances (i.e., alkylphenols, hormones, pharmaceuticals, pesticides, PCBs, bisphenol A) in POCIS fractions and effluent extracts. However, mass-balance calculations showed that the analyzed compounds explained only 0.03% and 1.4% of biological activity measured in POCIS and STW samples, respectively. In effluents, bisphenol A and 4-tert-octylphenol were identified as main contributors of instrumentally derived PXR activities. Finally, the PXR bioassay provided complementary information as compared to estrogenic, androgenic, and dioxin-like activity measured in these samples. This study shows the usefulness of HG5LN-hPXR cells to detect PXR-active compounds in water samples, and further investigation will be necessary to identify the detected active compounds. PMID- 20024650 TI - Apparent motion during saccadic suppression periods. AB - Sensitivity to many visual stimuli, and, in particular, image displacement, is reduced during a change in fixation (saccade) compared to when the eye is still. In these experiments, we studied the sensitivity of observers to ecologically relevant image translations of large, complex, real world scenes either during horizontal saccades or during fixation. In the first experiment, we found that such displacements were much less detectable during saccades than during fixation. Qualitatively, even when trans-saccadic scene changes were detectable, they were less salient and appeared slower than equivalent changes in the absence of a saccade. Two further experiments followed up on this observation and estimated the perceived magnitude of trans-saccadic apparent motion using a two interval forced-choice procedure (Experiment 2) and a magnitude estimation procedure (Experiment 3). Both experiments suggest that trans-saccadic displacements were perceived as smaller than equivalent inter-saccadic displacements. We conclude that during saccades, the magnitude of the apparent motion signal is attenuated as well as its detectability. PMID- 20024651 TI - Egocentric and allocentric reference frames for catching a falling object. AB - When programming movement, one must account for gravitational acceleration. This is particularly important when catching a falling object because the task requires a precise estimate of time-to-contact. Knowledge of gravity's effects is intimately linked to our definition of 'up' and 'down'. Both directions can be described in an allocentric reference frame, based on visual and/or gravitational cues, or in an egocentric reference frame in which the body axis is taken as vertical. To test which frame humans use to predict gravity's effect, we asked participants to intercept virtual balls approaching from above or below with artificially controlled acceleration that could be congruent or not with gravity. To dissociate between these frames, subjects were seated upright (trunk parallel to gravity) or lying down (body axis orthogonal to the gravitational axis). We report data in line with the use of an allocentric reference frame and discuss its relevance depending on available gravity-related cues. PMID- 20024652 TI - Chronic intermittent materno-fetal hyperoxygenation in late gestation may improve on hypoplastic cardiovascular structures associated with cardiac malformations in human fetuses. AB - Hypoplasia of cardiovascular structures is a common finding in fetuses with cardiac malformations. Materno-fetal hyperoxygenation (HO) during late gestation promotes venous return to the fetal heart. This analysis in human fetuses sought to define whether this "loading" effect might improve hypoplastic cardiovascular dimensions. Fifteen late-gestation fetuses presented with varying degrees of hypoplastic cardiovascular structures. In these cases, chronic intermittent materno-fetal HO was administered during periods ranging from 8 to 33 days. Cardiac measurements were taken before and at the end of treatment and translated into Z-scores as well as plotted on normal growth charts. During the treatment period, chronic intermittent materno-fetal HO was associated with improved dimensions of >or=1 hypoplastic cardiovascular structures in most fetuses. However, in some cases, the effect of HO was neutralized or impaired by the presence of ventricular septal defects as well as obstructions to ventricular filling or emptying. Chronic intermittent materno-fetal HO near term may be associated with improvements of hypoplastic cardiovascular dimensions in fetuses with a spectrum of cardiac malformations. This effect may facilitate postnatal treatment and improve prognosis in suitable cases. PMID- 20024653 TI - Improved classification of coronary artery abnormalities based only on coronary artery z-scores after Kawasaki disease. AB - Competing definitions and classifications of coronary artery abnormalities (CAAs) after Kawasaki disease (KD) have been arbitrarily defined based on clinical experience. We sought to propose a classification system for CAAs based only on coronary artery z-scores. All echocardiograms performed between 1990 and 2007 on patients with a previous history of KD were reviewed. Coronary artery luminal dimensions were converted to body-surface-area-adjusted z-scores and compared to current classification systems. A total of 1356 patients with a previous history of KD underwent 4379 echocardiograms. There was important overlap in the distributions of coronary artery z-scores between the different CAA classes as defined by the American Heart Association (AHA). The AHA classification underestimated the severity of CAAs in 19-32% of small CAAs and 35-78% of medium CAAs. We determined the optimal definition of CAA to be small if the z-score is >or=2.5 to <5.0, large if the z-score is >or=5.0 to <10.0, and giant if the z score is >or=10.0. This classification seems to appropriately apply to the circumflex branch despite a lack of normal values for this branch. The current AHA classification might not accurately classify CAAs in KD patients. Accurate classification is important for defining management and prognosis consistently across patient age and size. PMID- 20024654 TI - Harnessing ecosystem models and multi-criteria decision analysis for the support of forest management. AB - The decision-making environment in forest management (FM) has changed drastically during the last decades. Forest management planning is facing increasing complexity due to a widening portfolio of forest goods and services, a societal demand for a rational, transparent decision process and rising uncertainties concerning future environmental conditions (e.g., climate change). Methodological responses to these challenges include an intensified use of ecosystem models to provide an enriched, quantitative information base for FM planning. Furthermore, multi-criteria methods are increasingly used to amalgamate information, preferences, expert judgments and value expressions, in support of the participatory and communicative dimensions of modern forestry. Although the potential of combining these two approaches has been demonstrated in a number of studies, methodological aspects in interfacing forest ecosystem models (FEM) and multi-criteria decision analysis (MCDA) are scarcely addressed explicitly. In this contribution we review the state of the art in FEM and MCDA in the context of FM planning and highlight some of the crucial issues when combining ecosystem and preference modeling. We discuss issues and requirements in selecting approaches suitable for supporting FM planning problems from the growing body of FEM and MCDA concepts. We furthermore identify two major challenges in a harmonized application of FEM-MCDA: (i) the design and implementation of an indicator-based analysis framework capturing ecological and social aspects and their interactions relevant for the decision process, and (ii) holistic information management that supports consistent use of different information sources, provides meta-information as well as information on uncertainties throughout the planning process. PMID- 20024655 TI - Diversity and community structure of archaea in deep subsurface sediments from the tropical Western pacific. AB - Archaeal 16S rRNA gene clone libraries using PCR amplicons from eight different layers of the MD06-3051 core were obtained from the tropical Western Pacific sediments. A total of 768 clones were randomly selected, and 264 representative clones were sequenced by restriction fragment length polymorphism. Finally, 719 valid clones and 104 operational taxonomic units were identified after chimera check and > or =97% similarity analysis. The phylogenetic analysis of 16S rDNA sequences obtained from sediment samples were very diverse and showed stratification with depth. Majority of the members were most closely related to uncultivated groups and physiologically uncharacterized assemblages. All phylotypes were affiliated with Crenarchaeota (76%) and Euryarchaeota (24%), respectively. Deep-sea archaeal group (DSAG, 41% of total clones) and miscellaneous crenarchaeotic group (MCG, 29% of total clones) belonging to Crenarchaeota were the most predominant archaeal 16S rDNA phylotypes in clone libraries. Phylotypes in this study shared high similarity with those in subsurface sediments from Peru Margin sites, which indicated that different geographical zones might host similar members of archaeal populations based on similar sedimentary environments. In our study, members of DSAG and MCG seemed to dominate certain layers of the nonhydrate sediments, suggesting a wide ecophysiological adaptation than previously appreciated. The spatial distribution and community structure of these groups might vary with the different geochemical gradients of the environment. PMID- 20024656 TI - A transcriptional "Scream" early response of E. coli prey to predatory invasion by Bdellovibrio. AB - We have transcriptionally profiled the genes differentially expressed in E. coli prey cells when predatorily attacked by Bdellovibrio bacteriovorus just prior to prey cell killing. This is a brief, approximately 20-25 min period when the prey cell is still alive but contains a Bdellovibrio cell in its periplasm or attached to and penetrating its outer membrane. Total RNA was harvested and labelled 15 min after initiating a semi-synchronous infection with an excess of Bdellovibrio preying upon E. coli and hybridised to a macroarray spotted with all predicted ORFs of E. coli. SAM analysis and t-tests were performed on the resulting data and 126 E. coli genes were found to be significantly differentially regulated by the prey upon attack by Bdellovibrio. The results were confirmed by QRT-PCR. Amongst the prey genes upregulated were a variety of general stress response genes, potentially "selfish" genes within or near prophages and transposable elements, and genes responding to damage in the periplasm and osmotic stress. Essentially, the presence of the invading Bdellovibrio and the resulting damage to the prey cell elicited a small "transcriptional scream", but seemingly no specific defensive mechanism with which to counter the Bdellovibrio attack. This supports other studies which do not find Bdellovibrio resistance responses in prey, and bodes well for its use as a "living antibiotic". PMID- 20024657 TI - Proton beam therapy following resection for childhood ependymoma. AB - INTRODUCTION: Proton radiation therapy is a form of radiation with physical properties that can provide an advantage in normal tissue sparing compared to the more commonly used photon therapy. DISCUSSION: The greatest benefit may be for young patients with tumors requiring relatively high doses of radiation and adjacent to critical structures. Children with central nervous system ependymomas meet these criteria and have a very high likelihood of benefit from proton radiation. At present, proton radiation is limited to select centers. However, knowledge of many of the physical advantages of proton therapy has spread rapidly over the past several years, and its availability is becoming more widespread. PMID- 20024658 TI - Perforation holes in ventricular catheters--is less more? AB - OBJECTIVE: Obstruction is a common cause of cerebrospinal fluid (CSF) shunt failure. Risk factors for proximal obstructive malfunction are suboptimal ventricular catheter positioning and slit-like ventricles. A new ventricular catheter design to decrease risk of obstruction was evaluated. METHODS: A review of histopathological tissue investigation from occluded ventricular catheters (n = 70) was performed. A new ventricular catheter design was realized with six perforation holes. These catheters were compared to regular catheters (16 holes, Miethke, Aesculap) for flow characteristics using ink studies and flow velocity at hydrostatic pressure levels from 14 to 2 cmH(2)O in an experimental setup. The six-hole catheters were implanted in hydrocephalic patients with slit-like ventricles (n = 55). A follow-up was performed to evaluate the need of catheter revisions. RESULTS: Histological evaluation showed that obstructive tissue involved 43-60% extraventricular tissue, including gliosis, connective and inflammatory cells. In flow characteristic studies, the 16-hole catheters showed that only proximal perforations are of functional relevance. For six-hole catheters, all perforations were shown to be relevant with remaining reserve capacity. Flow velocity however showed no significant differences between six and 16 perforations. The six-hole catheter was implanted in 55 patients with a mean follow-up period of 15 +/- 9 months. A total of 12 catheters were explanted, revealing an overall survival proportion of 77.4%. CONCLUSION: In narrow ventricles, we assume that catheter perforations that are located also in the tissue might be a risk for CSF shunt obstruction. Fewer amounts of perforations in the catheters with equal flow features might decrease this risk when catheters can be implanted with adequate precision. PMID- 20024659 TI - Ependymal cells: biology and pathology. AB - The literature was reviewed to summarize the current understanding of the role of ciliated ependymal cells in the mammalian brain. Previous reviews were summarized. Publications from the past 10 years highlight interactions between ependymal cells and the subventricular zone and the possible role of restricted ependymal populations in neurogenesis. Ependymal cells provide trophic support and possibly metabolic support for progenitor cells. Channel proteins such as aquaporins may be important for determining water fluxes at the ventricle wall. The junctional and anchoring proteins are now fairly well understood, as are proteins related to cilia function. Defects in ependymal adhesion and cilia function can cause hydrocephalus through several different mechanisms, one possibility being loss of patency of the cerebral aqueduct. Ependymal cells are susceptible to infection by a wide range of common viruses; while they may act as a line of first defense, they eventually succumb to repeated attacks in long lived organisms. Ciliated ependymal cells are almost certainly important during brain development. However, the widespread absence of ependymal cells from the adult human lateral ventricles suggests that they may have only regionally restricted value in the mature brain of large size. PMID- 20024660 TI - New developments in reflux-associated cough. AB - Gastro-oesophageal reflux disease (GORD) is generally considered one of the three main causes of chronic cough, along with asthma and nasal disease. The diagnosis of GORD is often based upon a successful trial of anti-acid treatment however GORD is a complex condition taking many forms. Only recently have studies started to address the different types of GORD in patients with chronic cough and how these may infer the mechanisms linking these common conditions. GORD can be assessed in a number of ways; whilst endoscopy provides evidence of oesophagitis (i.e. erosive disease), 24-h ambulatory oesophageal pH monitoring may demonstrate abnormal oesophageal acid exposure in the absence of oesophageal damage (i.e. non erosive disease). The development of oesophageal impedance monitoring now allows the assessment of all reflux events (regardless of degree of acidity) and further classification of reflux by the proximal extension e.g. to upper oesophagus or even pharynx. Chronic cough patients may still be considered to have GORD if there is a significant temporal association between reflux events and coughing. Recent studies have examined the relationships between cough and reflux events, the roles of distal and proximal/pharyngeal reflux and also micro-aspiration in chronic cough patients. Increasing evidence suggests a significant proportion of patients display statistical associations between reflux and cough events, in the absence of an excessive numbers of reflux events either within or outside of the oesophagus. PMID- 20024662 TI - Expression level of Bmi-1 oncoprotein is associated with progression and prognosis in colon cancer. AB - PURPOSE: To investigate the clinicopathologic significance and predictive value of Bmi-1 expression in patients with colon cancer. METHODS: Bmi-1 expression was assessed by immunohistochemistry, PCR, and western blotting in specimens from 203 patients and by immunohistochemistry in 66 specimens of lymph node metastasis (LNM). RESULTS: Positive staining of Bmi-1 occurred in 7.9% (16/203), 66.5% (135/203), and 86.4% (57/66) of specimens from normal tissue, colon cancer, and LNM, respectively. Staining was significantly correlated with clinical stage, depth of invasion, nodal involvement, distant metastasis, and Ki67 level. Bmi-1 was upregulated at the transcriptional and translational levels. Patients with Bmi-1-positive localized tumors had a much lower 5-year disease-free survival (relative risk 2.919, P < 0.0001) and overall survival (relative risk 5.056, P < 0.0001). Bmi-1 immunoreactivity emerged as an independent prognostic factor in the multivariate analysis. CONCLUSIONS: We have shown that expression of Bmi-1 was elevated in colon cancer and might serve as an independent prognostic marker. PMID- 20024663 TI - Rigorous kinetic model considering positional specificity of lipase for enzymatic stepwise hydrolysis of triolein in biphasic oil-water system. AB - A rigorous kinetic model describing the stepwise triglyceride hydrolysis at the oil-water interface, based on the Ping Pong Bi Bi mechanism using suspended lipase having positional specificity, was constructed. The preference of the enzyme to cleave to the ester bonds at the edge and the center of the glycerol backbone of the substrates (tri-, di- or monoglyceride) was incorporated in the model. This model was applied to the experimental results for triolein hydrolysis using suspended Porcine pancreatic lipase (an sn-1,3 specific lipase) and Candida rugosa lipase (a non-specific lipase) in a biphasic oil-water system under various operating conditions. In order to discuss the model's advantages, other models that do not consider the positional specificity of the lipase were also applied to our experimental results. The model considering the positional specificity of the lipase gave results which fit better with the experimental data and described the effect of the initial enzyme concentration, the interfacial area, and the initial concentrations of triolein on the entire process of the stepwise triolein hydrolysis. This model also gives a good representation of the rate for cleaving the respective ester bonds of each substrate by each type of lipase. PMID- 20024665 TI - Photodynamic therapy as an adjunctive treatment for chronic periodontitis: a meta analysis. AB - Several antimicrobial strategies have been proposed in response to the alarming rise in antimicrobial resistance of periodontal pathogens. Antimicrobial photodynamic therapy (a-PDT) is a promising novel approach that has been used in several clinical applications including in the treatment of periodontal diseases. The aim of this review was to systematically investigate the effectiveness of a PDT as an adjuvant treatment for chronic periodontitis. The guidelines of the Quality of Reporting of Meta-analyses (QUOROM) conference statement were followed in the preparation of this meta-analysis. An electronic search for randomized controlled trials (RCTs) that investigated the combined use of scaling and root planing (SRP) and a-PDT in comparison with SRP alone was performed without language restriction up to 1 October 2008. RevMan 5.0 software was used to analyze the data. A random effects model was chosen and standardized mean differences with 95% confidence intervals were calculated for continuous data. Four RCTs were included. The use of a-PDT in conjunction with SRP was associated with significantly greater attachment gain (mean difference 0.29, 95% confidence interval 0.08 to 0.50, p=0.007), and greater reduction in probing depth (mean difference 0.11, 95% confidence interval -0.12 to 0.35, p=0.35) at 12 weeks. However, the changes in gingival recession showed slight differences. This review and meta-analysis supported the potential improvements in clinical attachment level and probing depth provided by the combined approach (SRP with a-PDT). Nevertheless, the findings of this review should be interpreted with caution given the small number of included studies. PMID- 20024664 TI - The Michel Benoist and Robert Mulholland yearly European Spine Journal Review: a survey of the "surgical and research" articles in the European Spine Journal, 2009. PMID- 20024666 TI - Social cost of heavy drinking and alcohol dependence in high-income countries. AB - OBJECTIVE: A comprehensive review of cost drivers associated with alcohol abuse, heavy drinking, and alcohol dependence for high-income countries was conducted. METHOD: The data from 14 identified cost studies were tabulated according to the potential direct and indirect cost drivers. The costs associated with alcohol abuse, alcohol dependence, and heavy drinking were calculated. RESULTS: The weighted average of the total societal cost due to alcohol abuse as percent gross domestic product (GDP)--purchasing power parity (PPP)--was 1.58%. The cost due to heavy drinking and/or alcohol dependence as percent GDP (PPP) was estimated to be 0.96%. CONCLUSIONS: On average, the alcohol-attributable indirect cost due to loss of productivity is more than the alcohol-attributable direct cost. Most of the countries seem to incur 1% or more of their GDP (PPP) as alcohol-attributable costs, which is a high toll for a single factor and an enormous burden on public health. The majority of alcohol-attributable costs incurred as a consequence of heavy drinking and/or alcohol dependence. Effective prevention and treatment measures should be implemented to reduce these costs. PMID- 20024667 TI - Small field of view imaging using wavelet encoding with 2 dimensional RF pulses and gradient echo: phantom results. AB - OBJECT: The objective of this work is to propose an imaging sequence based upon the wavelet encoding approach to provide MRI images free from folding artifacts, in the small field of view (FOV) regime, such as dynamic magnetic resonance imaging (MRI) studies. MATERIALS AND METHODS: The method consists of using a 2D spatially selective RF excitation pulse inserted into a gradient- echo pulse sequence to excite spins within a determined plane where wavelet encoding is achieved in one direction and slice selection is performed in the second direction. Wavelet encoding allows for spatially localized excitation and consequently restricts the spins excited within a reduced FOV. It consists of varying, according to a predetermined scheme, the width and position of the profile of the so-called fast RF pulse of the 2D RF excitation pulse, to obey wavelet encoding translation and dilation conditions. This sequence is implemented on a 3 Tesla whole body Siemens scanner. RESULTS: Compared to Fourier encoding, the proposed technique tested on phantoms with different shapes and structures, is able to provide gradient-echo reduced FOV images free from aliased signals. CONCLUSION: Wavelet encoding is suitable for small FOV imaging in dynamic MRI studies. PMID- 20024668 TI - Combining RF encoding with parallel imaging: a simulation study. AB - OBJECT: The aim of this work was to investigate combining spatial encoding by radio frequency (RF) excitation with conventional parallel imaging (PI) methods to determine whether this could improve overall imaging performance. MATERIALS AND METHODS: A simulation framework was developed to predict imaging performance for regular, central and random under-sampled parallel imaging methods augmented by RF spatial signal modulation. Optimisation methods were used to find the RF modulation patterns that produce optimal image reconstruction using the condition number of the PI encoding matrix as a quality metric. The diverse patterns of raw data sampling produced were compared using a measure of data uniformity across k space. RESULTS: Regular under-sampling of k-space provided the best reconstruction quality. When other under-sampling schemes were employed then RF modulation could be used to improve reconstruction, with the optimum achieved by redistributing the signal in k-space to return to regular sub-sampling. For all tested under-sampling patterns, no further improvements in image quality were attained. CONCLUSION: Using the simulation framework and metrics described the interaction of different spatial encoding approaches could be investigated. Regular sub-sampling provided optimal reconstruction, independent of whether the spatial encoding was achieved by gradients only or a combination of gradient and RF. PMID- 20024669 TI - Specific induction of TaAAPT1, an ER- and Golgi-localized ECPT-type aminoalcoholphosphotransferase, results in preferential accumulation of the phosphatidylethanolamine membrane phospholipid during cold acclimation in wheat. AB - Cold acclimation requires substantial alteration in membrane property. In contrast to well-documented fatty acid unsaturation during cold acclimation, changes in phospholipid biosynthesis during cold acclimation are less understood. Here, we isolated and characterized two aminoalcoholphosphotransferase (AAPT) cDNAs, TaAAPT1 and TaAAPT2, from wheat. AAPTs utilize diacylglycerols and CDP choline/ethanolamine as substrates and catalyze the final step of the CDP choline/ethanolamine pathway for phosphatidylcholine (PC)/phosphatidylethanolamine (PE) synthesis, respectively. Functionality of TaAAPT1 and TaAAPT2 was demonstrated by heterologous expression in a yeast cpt1Delta ept1Delta double mutant that lacks both AAPT activities. Detailed characterization of AAPT activities from the transformed mutant cells indicated that TaAAPT1 is an ECPT-type enzyme with higher ethanolamine phosphotransferase (EPT) activity than choline phosphotransferase (CPT) activity, while TaAAPT2 is a CEPT-type with the opposite substrate preference. Transient expression of GFP fused TaAAPT1 and TaAAPT2 proteins in wheat and onion cells indicated they are localized to both the endoplasmic reticulum and Golgi apparatus, suggesting that the final synthesis of PE and PC via the CDP-choline/ethanolamine pathway occurs in these organella. Quantitative PCR analyses revealed that TaAAPT1 expression is strongly induced by cold, while TaAAPT2 was constitutively expressed at lower levels. Measurement of phospholipid content in wheat leaves indicated that PE is more prominently increased in response to cold than PC and accordingly PE/PC ratio increased from 0.385 to 0.530 during 14 days of cold acclimation. Together, these data suggested that an increase in the PE/PC ratio during cold acclimation is regulated at the final step of the biosynthetic pathway. PMID- 20024670 TI - PLGA microparticles in respirable sizes enhance an in vitro T cell response to recombinant Mycobacterium tuberculosis antigen TB10.4-Ag85B. AB - PURPOSE: To study the use of poly (lactide-co-glycolide) (PLGA) microparticles in respirable sizes as carriers for recombinant tuberculosis (TB) antigen, TB10.4 Ag85B, with the ultimate goal of pulmonary delivery as vaccine for the prevention of TB. MATERIALS AND METHODS: Recombinant TB antigens were purified from E. coli by FPLC and encapsulated into PLGA microparticles by emulsion/spray-drying. Spray drying condition was optimized by half-factorial design. Microparticles encapsulating TB antigens were assessed for their ability to deliver antigens to macrophages for subsequent presentation by employing an in vitro antigen presentation assay specific to an Ag85B epitope. RESULTS: Spray-drying condition was optimized to prepare PLGA microparticles suitable for pulmonary delivery (aerodynamic diameter of 3.3 microm). Antigen release from particles exhibited an initial burst release followed by sustained release up to 10 days. Antigens encapsulated into PLGA microparticles induced much stronger interleukin-2 secretion in a T-lymphocyte assay compared to antigen solutions for three particle formulations. Macrophages pulsed with PLGA-MDP-TB10.4-Ag85B demonstrated extended epitope presentation. CONCLUSION: PLGA microparticles in respirable sizes were effective in delivering recombinant TB10.4-Ag85B in an immunologically relevant manner to macrophages. These results set the foundation for further investigation into the potential use of PLGA particles for pulmonary delivery of vaccines to prevent Mycobacterium tuberculosis infection. PMID- 20024671 TI - The association between conduct problems and maltreatment: testing genetic and environmental mediation. AB - It is often assumed that childhood maltreatment causes conduct problems via an environmentally mediated process. However, the association may be due alternatively to either a nonpassive gene-environment correlation, in which parents react to children's genetically-influenced conduct problems by maltreating them, or a passive gene-environment correlation, in which parents' tendency to engage in maltreatment and children's conduct problems are both influenced by a hereditary vulnerability to antisocial behavior (i.e. genetic mediation). The present study estimated the contribution of these processes to the association between maltreatment and conduct problems. Bivariate behavior genetic analyses were conducted on approximately 1,650 twin and sibling pairs drawn from a large longitudinal study of adolescent health (Add Health). The correlation between maltreatment and conduct problems was small; much of the association between maltreatment and conduct problems was due to a nonpassive gene-environment correlation. Results were more consistent with the hypothesis that parents respond to children's genetically-influenced conduct problems by maltreating them than the hypothesis that maltreatment causes conduct problems. PMID- 20024672 TI - A developmental examination of story recall and coherence among children with ADHD. AB - This study investigated developmental differences in story recall in children with attention-deficit/hyperactivity disorder (ADHD), N = 57 (77.2% male) and their comparison peers, N = 98 (61.2% male). Children at the ages of 4-6 or 7-9 completed a free recall immediately after viewing each of two televised stories, once in the presence of toys during viewing and once in their absence. This procedure was repeated with new stories 21 months later. Comparison children recalled more story events and showed a greater sensitivity to the thematic importance of the story events than did children with ADHD, a pattern that remained stable over time. Older comparison children showed a dramatic increase over time in the global coherence of their narrations, whereas the older children with ADHD showed limited improvement over time. The implications of these findings for academic performance and the possible need for remediation are discussed. PMID- 20024673 TI - Synemin isoforms in astroglial and neuronal cells from human central nervous system. AB - The intermediate filament (IF) synemin gene encodes three IF proteins (H 180, M 150, L 41 kDa) with overlapping distributions. Synemin M was present early with vimentin and nestin. Synemin H was found later in the nervous system and mesodermic derivatives concomitantly with angiogenesis and the migration of neural crest cells. Synemin L appeared later in neurons. A series of in vitro cell cultures were done to identify the linkage between synemin isoforms and specific cell types of the central nervous system (CNS). The neurons and glia from the brains of humans and rats were cultured and double immunostaining done with antibodies against the H/M or L synemin isoforms and neural cell types (betaIII-tubulin or NeuN) or astrocyte intermediate filaments (GFAP or vimentin). In neurons of the CNS, synemin H/M were co-expressed with GFAP, vimentin or nestin in glial cells, whereas synemin L was found in neurons. PMID- 20024674 TI - Pulmonary adenomatosis and maedi-visna in Ethiopian central highland sheep: a microscopic study. AB - Microscopic examination of pneumonic lungs of the Ethiopian highland sheep (n = 35) was made and compared with the pneumonic lungs from ten sheep and 66 goats from the lowlands. Lesions compatible with sheep pulmonary adenomatosis (SPA; 8/35, 22.8%), and maedi-visna (MV; 9/35, 25.7%) were recorded only in sheep from the central highlands. Interstitial pneumonia (43.2%), bronchopneumonia (35.1%), and verminous pneumonia (6.3%) were recorded in both sheep and goats from the high- and the lowlands. SPA was documented for the first time in sheep from Ethiopia in this report. We believe that MV and SPA were introduced into Ethiopia through importation of exotic sheep. These infections should be considered in dealing with the diagnosis of respiratory diseases in all the sheep breeds in the central highlands and in the exotic and the crossbred sheep in the other parts of the country. PMID- 20024675 TI - Colloids as a sink for certain pharmaceuticals in the aquatic environment. AB - BACKGROUND, AIM, AND SCOPE: The occurrence and fate of pharmaceuticals in the aquatic environment is recognized as one of the emerging issues in environmental chemistry and as a matter of public concern. Existing data tend to focus on the concentrations of pharmaceuticals in the aqueous phase, with limited studies on their concentrations in particulate phase such as sediments. Furthermore, current water quality monitoring does not differentiate between soluble and colloidal phases in water samples, hindering our understanding of the bioavailability and bioaccumulation of pharmaceuticals in aquatic organisms. In this study, an investigation was conducted into the concentrations and phase association (soluble, colloidal, suspended particulate matter or SPM) of selected pharmaceuticals (propranolol, sulfamethoxazole, meberverine, thioridazine, carbamazepine, tamoxifen, indomethacine, diclofenac, and meclofenamic acid) in river water, effluents from sewage treatment works (STW), and groundwater in the UK. MATERIALS AND METHODS: The occurrence and phase association of selected pharmaceuticals propranolol, sulfamethoxazole, meberverine, thioridazine, carbamazepine, tamoxifen, indomethacine, diclofenac, and meclofenamic acid in contrasting aquatic environments (river, sewage effluent, and groundwater) were studied. Colloids were isolated by cross-flow ultrafiltration (CFUF). Water samples were extracted by solid-phase extraction (SPE), while SPM was extracted by microwave. All sample extracts were analyzed by liquid chromatography-tandem mass spectrometry (LC-MS/MS) in the multiple reaction monitoring. RESULTS AND DISCUSSION: Five compounds propranolol, sulfamethoxazole, carbamazepine, indomethacine, and diclofenac were detected in all samples, with carbamazepine showing the highest concentrations in all phases. The highest concentrations of these compounds were detected in STW effluents, confirming STW as a key source of these compounds in the aquatic environments. The calculation of partition coefficients of pharmaceuticals between SPM and filtrate (observed partition coefficients, Kobsp, Kobsoc), between SPM and soluble phase (intrinsic partition coefficients, Kintp, Kintoc), and between colloids and soluble phase (Kcoc) showed that intrinsic partition coefficients (Kintp, Kintoc) are between 25% and 96%, and between 18% and 82% higher than relevant observed partition coefficients values, and are much less variable. Secondly, Kcoc values are 3-4 orders of magnitude greater than Kintoc values, indicating that aquatic colloids are substantially more powerful sorbents for accumulating pharmaceuticals than sediments. Furthermore, mass balance calculations of pharmaceutical concentrations demonstrate that between 23% and 70% of propranolol, 17-62% of sulfamethoxazole, 7-58% of carbamazepine, 19-84% of indomethacine, and 9-74% of diclofenac are present in the colloidal phase. CONCLUSIONS: The results provide direct evidence that sorption to colloids provides an important sink for the pharmaceuticals in the aquatic environment. Such strong pharmaceutical/colloid interactions may provide a long-term storage of pharmaceuticals, hence, increasing their persistence while reducing their bioavailability in the environment. RECOMMENDATIONS AND PERSPECTIVES: Pharmaceutical compounds have been detected not only in the aqueous phase but also in suspended particles; it is important, therefore, to have a holistic approach in future environmental fate investigation of pharmaceuticals. For example, more research is needed to assess the storage and long-term record of pharmaceutical residues in aquatic sediments by which benthic organisms will be most affected. Aquatic colloids have been shown to account for the accumulation of major fractions of total pharmaceutical concentrations in the aquatic environment, demonstrating unequivocally the importance of aquatic colloids as a sink for such residues in the aquatic systems. As aquatic colloids are abundant, ubiquitous, and highly powerful sorbents, they are expected to influence the bioavailability and bioaccumulation of such chemicals by aquatic organisms. It is therefore critical for colloids to be incorporated into water quality models for prediction and risk assessment purposes. PMID- 20024676 TI - Fractionation and bioavailability of Cu in soil remediated by EDTA leaching and processed by earthworms (Lumbricus terrestris L.). AB - BACKGROUND, AIM, AND SCOPE: Soil remediation with ethylenediamine tetraacetic acid (EDTA) leaching is capable of removing only part of the total metal concentration in the soil, mostly the labile, bioavailable metal species (metal bioavailability stripping). However, reintroduction of remediated soil in the environment exposes the soil to various environmental factors, which could potentially shift nonlabile residual metals back to labile bioavailable forms. We studied the effect of autochthonous earthworm species as model biotic environmental factor on the fractionation and bioavailability of Cu residual in soil after remediation. MATERIALS AND METHODS: We used soil from a 50-year-old vineyard regularly managed and treated with CuSO(4)*5H(2)O (Bordeaux mixture) as fungicide. Soil containing 400 mg kg(-1) of Cu was leached with total 15 mmol kg( 1) EDTA. Remediated and nonremediated soil was processed by fully clitellated adult specimens of Lumbricus terrestris L., a prevailing autochthonous soil earthworm species. Cu fractionation, phytoavailability, and oral-bioavailability in processed and nonprocessed soil were determined using six-step sequential extraction, extraction with diethylenediamine pentaacetic acid, and in vitro physiologically based extraction test, respectively. RESULTS: EDTA leaching removed 41% of the pseudototal Cu, mostly from the soil Fe- and Mn-oxides, carbonates, and organic matter. A 2.7-fold decrease in Cu phytoavailability and a 4.4- and 2.8-fold decrease in Cu oral-bioavailability in the stomach and small intestine fractions, respectively, were achieved after remediation. In nonremediated soil, earthworms increased the share of nonlabile Cu in residual soil fraction, while in remediated soil they increased the share of Cu bound to carbonates. A statistically significant 1.1- and 1.7-fold increase in Cu phytoavailability and intestinal oral-bioavailability, respectively, was observed in earthworm processed remediated soil. DISCUSSION: Cu occurs in various soil "pools" of different solubilities with different chemical characteristics and consequently different functions. By removing the labile part of the metals from the soil during remediation, we disrupt the chemical equilibrium; the nonlabile residual metals left in soil after remediation might become more labile in time in tendency to re-establish that equilibrium. Earthworms alter the physical and chemical properties of soil affecting consequently the fractionation of metals. The increase in earthworm's gut pH due to the excretion of ammonia and/or calcium carbonate into the intestine could lead to the transbounding of metals into the carbonate fraction. However, their activity in remediated soil increased Cu phytoavailability and intestinal oral-bioavailability, and it would, therefore, be improper to generalize the influence of earthworms on metal availability in soil. CONCLUSIONS: The results presented here show that residual Cu in remediated soil is affected by environmental factors such as earthworms, which should be considered in evaluating the effect of Cu polluted soil remediation. RECOMMENDATIONS AND PERSPECTIVES: Information on the behavior of residual metals in soil after its remediation is surprisingly scarce. The development of new effective remediation techniques should imply also the evaluation of postremediation effects on remediated soil. The results presented in this work indicate a possible tool for assessing the effect of biotic environmental factors on residual metals left in soil after its remediation. PMID- 20024677 TI - MFalpha signal peptide enhances the expression of cellulase eg1 gene in yeast. AB - Ethanol production from lignocellulose by recombinant yeast with high level expression of heterologous cellulase genes has been a major anticipation. The native secretion signal sequence of the cellulase endoglucanase I (eg1) gene was replaced by Saccharomyces cerevisiae mating factor alpha prepro-leader sequence (MFalpha). The transformants containing native secretion signal (Y(1)) and MFalpha secretion signal (Y(2)) were characterized with respect to gene expression and growth on cellulose substrate. Increased enzyme activity and cellulose utilization were observed. The enzyme activity of Y(2) was 0.084 U/ml, 61.5% higher than Y(1) (0.052 U/ml). The sufficiency parameter (S value) was raised from 0.6 to 0.84. MFalpha signal peptide was more efficient than the native signal peptide of eg1 gene, suggesting that signal peptide replacement is an efficient way to enhance the cellulase expression level in yeast, for cellulose-derived ethanol production. PMID- 20024678 TI - Clinical update of the AS04-adjuvanted human papillomavirus-16/18 cervical cancer vaccine, Cervarix. AB - Persistent infection with human papillomavirus (HPV) is a necessary cause of cervical cancer, resulting annually in 274,000 deaths worldwide. Two prophylactic HPV vaccines are licensed in >100 countries, and immunization programs in young, adolescent girls have been widely implemented. HPV-16/18 AS04-adjuvanted vaccine (Cervarix; GlaxoSmithKline Biologicals, Rixensart, Belgium) has demonstrated type specific protection against the five most frequent cancer-causing types (16, 18, 31, 33, and 45) that are responsible for 82% of invasive cervical cancers globally. Cervarix has demonstrated efficacy against HPV-45, which is the third most common HPV type in cervical cancer and adenocarcinoma. Final results of a large phase 3 trial recently showed Cervarix substantially reduced the overall burden of cervical precancerous lesions (cervical intraepithelial neoplasia 2+) by 70.2% in an HPV-naive population approximating young girls prior to sexual debut, the target of most current vaccination programs. Protection offered by Cervarix against nonvaccine types (mainly 31, 33, and 45) might potentially allow for 11%-16% additional protection against cervical cancers, compared to a vaccine only offering protection against HPV-16/18. Another recent study directly compared the antibody response of Cervarix to that of quadrivalent HPV-6/11/16/18 vaccine (Gardasil; Merck, Whitehouse Station, NJ, USA). Cervarix induced significantly superior neutralizing antibody levels as compared with Gardasil for HPV-16 and HPV-18 in all age groups studied. This may translate into more women having detectable (neutralizing) antibodies in cervicovaginal secretions for HPV 16 and HPV-18 after vaccination with Cervarix when compared with Gardasil. Cervarix induced significantly higher frequencies of antigen-specific memory B cells and T-cells in responders for HPV-16 and HPV-18 as compared with Gardasil. Cervarix continues to show sustained high levels of total and neutralizing antibodies for HPV-16 and HPV-18, 7.3 years after vaccination. This is associated with high efficacy and no breakthrough cases in the HPV-naive population, and is the longest duration follow-up for safety, immunogenicity, and efficacy for any licensed HPV vaccine to date. PMID- 20024679 TI - LigaSure versus clamp-and-tie in thyroidectomy: a single-center experience. AB - PURPOSE: LigaSure (ValleyLab, CO, USA) is a bipolar diathermy system that seals vessels with reduced thermal spread. The device has been used successfully in abdominal surgery and has been introduced as a new method for hemostasis during thyroidectomy. This study compares the efficacy and advantages of LigaSure, when used for thyroidectomy in benign nodular thyroid disease, with the conventional clamp-and-tie technique. METHODS: Between December 2003 and December 2008, 410 patients underwent thyroid surgery (333 with the conventional clamp-and-tie technique and 77 with LigaSure). Nine patients did not sign the ethical approval document and so were excluded from the study; the results of 68 LigaSure patients were evaluated. The main outcomes measured were age, sex, operating time, postoperative hypocalcemia, recurrent laryngeal nerve palsy, and drain use. Statistical analysis consisted of the chi-squared test and Student's t-test. RESULTS: The mean operating time was significantly shorter in the LigaSure group by 24.1+/-1.7 minutes for a total thyroidectomy (P<0.0001) and 7.6+/-0.1 minutes for one side total plus other side sub- total thyroidectomy (P<0.0001). Bleeding during surgery was significantly lower in the LigaSure group (P<0.0001). There were no significant differences between the study groups in terms of postoperative hypoparathyroidism, recurrent laryngeal nerve palsy, and postthyroidectomy hemorrhage. CONCLUSION: The LigaSure vessel sealer is a safe, effective alternative for thyroid surgery that reduces the overall operating time. However, its higher cost is a significant handicap for its use in surgery in developing countries. PMID- 20024680 TI - Amlodipine/valsartan/hydrochlorothiazide triple combination therapy in moderate/severe hypertension: Secondary analyses evaluating efficacy and safety. AB - INTRODUCTION: An 8-week trial of amlodipine/valsartan/hydrochlorothiazide (Aml/Val/HCTZ) for moderate or severe hypertension demonstrated more-pronounced blood pressure (BP)-lowering effects compared with dual-component therapies. To elucidate the effects of time and baseline BP on the observed responses, exploratory analyses were performed. METHODS: Patients aged 18-85 years with mean sitting systolic BP (MSSBP) 145 to <200 mmHg and mean sitting diastolic BP (MSDBP) 100 to <120 mmHg were randomized to Aml 10 mg/Val 320 mg/HCTZ 25 mg; Val 320 mg/HCTZ 25 mg; Aml 10 mg/Val 320 mg; or Aml 10 mg/HCTZ 25 mg. During the first 2 weeks, regimens were force-titrated in two stages. RESULTS: All least square mean reductions in MSSBP and MSDBP (baseline to Week 3 and end of study) were significantly greater with triple therapy than with each dual therapy in the overall population and the severe systolic subgroup (baseline MSSBP > or =180 mmHg; except vs. Aml 10 mg/Val 320 mg at Week 3). At Week 3, more patients on triple therapy achieved MSSBP reductions of > or =-60, > or =-50, > or =-40, > or =-30, and > or =-20 mmHg (2.5%, 9.7%, 23.2%, 46.9% and 74.5%, respectively) than those on dual therapy (1.1%-2%, 5.6%-5.9%, 14.5%-16.7%, 33.5%-39.1%, and 58.8% 65.5%, respectively); this was also true at study endpoint. End-of-study MSSBP reductions were greater in triple-therapy recipients who had higher (vs. lower) baseline MSSBPs. LSM reductions ranged from -27.2 mmHg for baseline MSSBP 145 to <150 mmHg, to > or =49.6 mmHg for baseline MSSBP > or =180 mmHg. All treatments were well tolerated regardless of baseline MSSBP. CONCLUSION: Aml 10 mg/Val 320 mg/HCTZ 25 mg triple therapy is highly effective in reducing BP compared with dual components early in therapy, and systolic BP-lowering effects were proportionate to hypertension severity. PMID- 20024681 TI - Myocardial bridges of the coronary arteries in the human fetal heart. AB - During the last century, many investigators reported on myocardial bridges in the adult human heart. In the present study, 39 human fetal hearts (the mean gestastional age was 30 weeks) were studied for myocardial bridging, and the results were correlated with adult data. Among the 39 (27 male and 12 female) fetal hearts studied, 26 bridges were observed on 18 fetal hearts (46.2%). Ten of the bridges had one myocardial bridge, whereas double myocardial bridges were observed in eight fetal hearts. The most frequent myocardial bridges were observed on the left anterior descending artery (LAD), which had 13 bridges (50%). Eight (30.7%) myocardial bridges were on the diagonal artery, and on the posterior descending artery there were five (19.3%). Myocardial bridges were not observed on the circumflex artery. The data presented in this study may provide potentially useful information for the preoperative evaluation of the newborn and may have a clinical implication for sudden fetal death. PMID- 20024682 TI - Novel fabrication of silver-coated glass capillaries for ready SERS-based detection of dissolved chemical species. AB - A novel method to deposit a highly surface-enhanced Raman scattering (SERS) active silver film onto the inside surface of a glass capillary is developed. Firstly, Ag sol was synthesized by the reaction of AgNO(3) with poly (ethylenimine) (PEI), and then toluene and benzenethiol (BT) were added into the sol. The mixture was flowed through the glass capillary to obtain the SERS-active Ag film-coated glass capillary. The SERS activity of the Ag-coated capillary was dependent on the amount of PEI and BT used. In addition, BT could be easily desorbed from the Ag surface by treating it with a borohydride solution, maintaining the initial SERS activity. The SERS enhancement factor at 632.8-nm excitation was estimated to be on the order of 10(6). The detection limits of adenine and dipicolinic acid were then as low as 1.0 x 10(-8) and 1.0 x 10(-7) M, respectively, based on an S/N ratio of 3. This clearly suggests that the Ag coated capillary is an invaluable device for the analysis of effluent chemicals by SERS. PMID- 20024683 TI - Analysis and occurrence of alkylphenolic compounds and estrogens in a European river basin and an evaluation of their importance as priority pollutants. AB - As a part of a project aiming to assess the potential toxicological effects of contaminants in aquatic ecosystems, the objective of this work was to determine the occurrence of several selected endocrine-disrupting compounds in water and sediment and to estimate the estrogenicity of the water. The study consisted of four sampling campaigns at seven sampling points in the lower Llobregat catchment area (NE Spain). Water and sediment samples underwent chemical target analysis for 19 steroid estrogens and alkylphenols, which are known to be endocrine disrupting compounds. In this study, the only estrogens detected in the water samples were estrone and estrone-sulfate, which were found at low levels (2-5 ng l(-1)). The alkylphenolic compound showing the highest concentrations was nonylphenol di-ether carboxylate (NP2EC), which was found at levels up to 30.62 microg l(-1) in water samples and 535 ng g(-1) in sediment samples. K (d) was determined for several alkylphenolic compounds and showed the expected trend of decreasing K (d) with increasing polarity. The concentrations of nonylphenol and octylphenol only exceeded the annual average of the European Union's environmental quality standards (EQS) in one sampling point. However, the calculated estrogenic potential surpassed the expected effect concentration in several sampling points, indicating a potential risk. Therefore, we recommend that future EQS include short-chain alkylphenol ethoxylates and carboxylates. PMID- 20024684 TI - Diversity and community structure of Archaea inhabiting the rhizoplane of two contrasting plants from an acidic bog. AB - Plant root exudates increase nutrient availability and influence microbial communities including archaeal members. We examined the archaeal community inhabiting the rhizoplane of two contrasting vascular plants, Dulichium arundinaceum and Sarracenia purpurea, from an acidic bog in upstate NY. Multiple archaeal 16S rRNA gene libraries showed that methanogenic Archaea were dominant in the rhizoplane of both plants. In addition, the community structure (evenness) of the rhizoplane was found markedly different from the bulk peat. The archaeal community in peat from the same site has been found dominated by the E2 group, meanwhile the rhizoplane communities on both plants were co-dominated by Methanosarcinaceae (MS), rice cluster (RC)-I, and E2. Complementary T-RFLP analysis confirmed the difference between bulk peat and rhizoplane, and further characterized the dominance pattern of MS, RC-I, and E2. In the rhizoplane, MS was dominant on both plants although as a less variable fraction in S. purpurea. RC-I was significantly more abundant than E2 on S. purpurea, while the opposite was observed on D. arundinaceum, suggesting a plant-specific enrichment. Also, the statistical analyses of T-RFLP data showed that although both plants overlap in their community structure, factors such as plant type, patch location, and time could explain nearly a third of the variability in the dataset. Other factors such as water table, plant replicate, and root depth had a low contribution to the observed variance. The results of this study illustrate the general effects of roots and the specific effects of plant types on their nearby archaeal communities which in bog-inhabiting plants were mainly composed by methanogenic groups. PMID- 20024686 TI - Relevance of pupil size in the clinical determination of retinal straylight on young healthy human eyes. AB - PURPOSE: To determine the impact of pupil size in the psychometric determination of intraocular straylight determined by the compensation comparison method in a population of young healthy adults. METHODS: Straylight was measured in 21 eyes from 11 young healthy subjects using the C-Quant (Oculus, Germany). Subjects were dilated with Phenylephrine 5%, and varying pupil size was simulated using three different diaphragms of 3, 5 and 6 mm in diameter. Three measurements were taken with each of the apertures in a randomized sequence. Mixed effects and repeated measures ANOVA was applied to test the significance of differences in straylight values for the fixed pupil diameters. Correlation analysis was applied to test relationships between the measurements for each fixed pupil diameter, and Bland Altmann plots displayed the distribution of differences. RESULTS: Significant differences between the straylight values obtained for the different fixed pupil diameters were not found. Correlation analysis did not show relationships between the values obtained for different diameters either (p > 0.05 in all cases). Bland Altmann plots show that average differences are around zero regardless of the actual value measured, but there is also a substantial individual variation, with differences going as far up as 0.5 Log(s) units in some cases. CONCLUSION: In healthy human eyes, straylight values measured with the Van den Berg straylightmeter are not affected significantly by the subject's pupil size, although individual variations exist implying that no control for pupil size is required when estimating average amounts of retinal straylight from samples of eyes, but is desirable for the assessment of longitudinal variations of straylight in individual subjects, especially when assessing effects of surgical procedures and/or lens designs. PMID- 20024685 TI - Microbial fuel cells and microbial ecology: applications in ruminant health and production research. AB - Microbial fuel cell (MFC) systems employ the catalytic activity of microbes to produce electricity from the oxidation of organic, and in some cases inorganic, substrates. MFC systems have been primarily explored for their use in bioremediation and bioenergy applications; however, these systems also offer a unique strategy for the cultivation of synergistic microbial communities. It has been hypothesized that the mechanism(s) of microbial electron transfer that enable electricity production in MFCs may be a cooperative strategy within mixed microbial consortia that is associated with, or is an alternative to, interspecies hydrogen (H(2)) transfer. Microbial fermentation processes and methanogenesis in ruminant animals are highly dependent on the consumption and production of H(2)in the rumen. Given the crucial role that H(2) plays in ruminant digestion, it is desirable to understand the microbial relationships that control H(2) partial pressures within the rumen; MFCs may serve as unique tools for studying this complex ecological system. Further, MFC systems offer a novel approach to studying biofilms that form under different redox conditions and may be applied to achieve a greater understanding of how microbial biofilms impact animal health. Here, we present a brief summary of the efforts made towards understanding rumen microbial ecology, microbial biofilms related to animal health, and how MFCs may be further applied in ruminant research. PMID- 20024687 TI - Selective retina therapy (SRT) in patients with geographic atrophy due to age related macular degeneration. AB - BACKGROUND: For geographic atrophy (GA) due to age-related macular degeneration (AMD) there is so far no approved treatment option. Usually, increased autofluorescence (AF) levels of different patterns adjacent to the atrophic area indicate lipofuscin-laden retinal pigment epithelium (RPE) cells at a high risk for apoptosis. Herein, SRT was used to selectively treat these cells to stimulate RPE proliferation, in order to reduce or ideally stop further growth of the atrophic area. MATERIAL AND METHODS: Six eyes of six patients with bilateral equally pronounced GA were treated by SRT, while the fellow eye served as control. Irradiation was performed using a prototype SRT laser (Medical Laser Center Lubeck, Nd:YLF laser; 527 nm; 200 ns/1.7 micros pulse duration; 30 repetitive pulses at 100 Hz). Test lesions with increasing energies were applied at the lower vessel arcade to determine the individual angiographic and ophthalmoscopic threshold radiant exposures. Treatment was then performed in the area of increased AF adjacent to the GA using energies between both thresholds. The GA progression rates of treated and fellow eyes were evaluated. RESULTS: After a 1-year follow-up, a progression of the atrophic area was observed in the treated eyes (0.7-8.0 mm(2)/yr, mean 3.0 mm(2)/yr; 46%/yr) whereas the progression rates of the fellow eyes were insignificantly lower (0.46-4.04 mm(2)/yr, mean 1.9 mm(2)/yr; 30%/yr; p = 0.134). The progression rate in the treated eyes of two patients increased significantly, while in the other four patients, the progression rates were nearly the same between both eyes. Moreover, one of these two eyes showed an unexpected RPE reaction after treatment, since all laser lesions led to RPE atrophy and thus an accelerated enlargement of the GA occurred. CONCLUSION: SRT in the hyperautofluorescent areas of GA was not able to stop or slow down the progression of GA. However, modified treatment strategies might be more promising, e.g. placing the spots outside the hyperautofluorescent areas where RPE apoptosis is postulated. Moreover, SRT studies on GA might be more successfully performed on specific subgroups of GA, based on autofluorescence and other findings. PMID- 20024689 TI - Endoscopically assisted release of the ulnar nerve for cubital tunnel syndrome. AB - PURPOSE: Recently, the simple decompression of the ulnar nerve has been advocated as the best surgical approach for the treatment of the cubital tunnel syndrome. Encouraged by the positive results observed with the use of the endoscopic approach for the treatment of the carpal tunnel syndrome, there have been reports about the use of endoscopes for decompression of the ulnar nerve at the level of the elbow since 1999. The objective of this study was to demonstrate the surgical results obtained with a simple and replicable technique employed for endoscopic release of the ulnar nerve in cases of cubital tunnel syndrome. METHODS: It was retrospectively studied thirteen patients who presented signs and symptoms of cubital tunnel syndrome and who were operated on by an endoscopically assisted technique, from 2007 to 2009. The approach included the use of a 0 degrees lens neuroendoscope usually employed for ventricular endoscopy. Preoperative clinical and electrophysiological data were collected and compared to those observed 6 months after the surgery. The Dellon's scale was used for rating the severity of the lesions, and the postoperative outcome was assessed based on the Bishop rating system. RESULTS: All procedures were completed successfully via the endoscopically assisted approach, and no surgery had to be converted to an open operation. Postoperatively, 76.9% of the cases were completely free of signs and symptoms (8 and 9 points on the Bishop scale), 15.3% presented with light complaints (7 points), and only one subject (7.6%) reached 5 points on the outcome scale. Complete normalization of the electrophysiological studies was also observed in seven patients, most of whom were classified preoperatively as Dellon's grades I and II, but three of whom were classified as grade III. Normalization of the sensory conduction studies was observed in ten cases, normalization of the motor conduction studies in six subjects, and in four patients, there was improvement in electromyographic parameters. CONCLUSIONS: The endoscopically assisted approach for decompression of the ulnar nerve at the level of the elbow is a minimally invasive technique that demonstrated surgical results similar to those reported via the open approach. It may have additional advantages such as the reduction of soft tissue manipulation, faster mobilization of the arm, and quicker return of the patients to their daily activities. PMID- 20024690 TI - Does molecular docking reveal alternative chemopreventive mechanism of activation of oxidoreductase by sulforaphane isothiocyanates? AB - Isothiocyanates (ITC) are well-known chemopreventive agents extracted from vegetables. This activity results from the activation of human oxidoreductase. In this letter, the uncompetitive activatory mechanism of ITC was investigated using docking and molecular dynamics simulations. This indicates that NAD(P)H:quinone oxidoreductase can efficiently improve enzyme-substrate recognition within the catalytic site if the ITC activator supports the interaction in the uncompetitive binding site. PMID- 20024691 TI - A novel Chk inhibitor, XL-844, increases human cancer cell radiosensitivity through promotion of mitotic catastrophe. AB - Check point kinases (Chk) play a major role in facilitating DNA repair upon radiation exposure. We tested the potency of a novel inhibitor of Chk1 and Chk2, XL-844 (provided by Exelixis Inc., CA, USA), to radiosensitize human cancer cells grown in culture and investigated the underlying mechanisms. HT-29 cells (a human colon cancer line) were exposed to XL-844, radiation, or both, and assessed for clonogenic cell survival. Treatment-dependent effects on phosphorylated forms of Chk proteins were assessed by Western blots. Further mechanistic investigations in HT-29 cells included cell cycle analysis by flowcytometry and assessment of DNA repair kinetics by immuno-cytochemistry (ICC) for nuclear appearance of the phosphorylated form of histone 2AX protein (gamma-H2AX) staining. Cells undergoing mitotic catastrophe were identified by irregular pattern of mitotic spindle markers alpha and gamma-tubulin staining by ICC. XL-844 enhanced radiosensitivity in a dose and schedule-dependent manner and the enhancement factor was 1.42 at 0.5 survival fraction. Mechanistically XL-844 abrogated radiation-induced Chk2 phosphorylation, induced pan-nuclear gamma-H2AX, and prolonged the presence of radiation-induced gamma-H2AX foci, and promoted mitotic catastrophe. In conclusion, our data showed that inhibition of Chk2 activity by XL-844 enhanced cancer cell radiosensitivity that was associated with inhibition of DNA repair and induction of mitotic catastrophe. PMID- 20024692 TI - Missense mutations in IHH impair Indian Hedgehog signaling in C3H10T1/2 cells: Implications for brachydactyly type A1, and new targets for Hedgehog signaling. AB - Heterozygous missense mutations in IHH result in Brachydactyly type A1 (BDA1; OMIM 112500), a condition characterized by the shortening of digits due to hypoplasia/aplasia of the middle phalanx. Indian Hedgehog signaling regulates the proliferation and differentiation of chondrocytes and is essential for endochondral bone formation. Analyses of activated IHH signaling in C3H10T1/2 cells showed that three BDA1-associated mutations (p.E95K, p.D100E and p.E131K) severely impaired the induction of targets such as Ptch1 and Gli1. However, this was not a complete loss of function, suggesting that these mutations may affect the interaction with the receptor PTCH1 or its partners, with an impact on the induction potency. From comparative microarray expression analyses and quantitative real-time PCR, we identified three additional targets, Sostdc1, Penk1 and Igfbp5, which were also severely affected. Penk1 and Igfbp5 were confirmed to be regulated by GLI1, while the induction of Sostdc1 by IHH is independent of GLI1. SOSTDC1 is a BMP antagonist, and altered BMP signaling is known to affect digit formation. The role of Penk1 and Igfbp5 in skeletogenesis is not known. However, we have shown that both Penk1 and Igfbp5 are expressed in the interzone region of the developing joint of mouse digits, providing another link for a role for IHH signaling in the formation of the distal digits. PMID- 20024693 TI - Steroid 11-beta hydroxylase deficiency caused by compound heterozygosity for a novel mutation in intron 7 (IVS 7 DS+4A to G) in one CYP11B1 allele and R448H in exon 8 in the other. AB - Congenital adrenal hyperplasia (CAH) due to steroid 11-beta hydroxylase deficiency (11beta-OHD) is a rare genetic disorder of steroidogenesis transmitted as an autosomal recessive trait. We describe a new case of 11beta-OHD CAH caused by compound heterozygosity for a novel mutation in intron 7 and previously described mutation in exon 8 of CYP 11B1 gene. A 2.5-year-old boy of Croatian descent presented with accelerated growth and bone age, borderline hypertension, and pseudoprecocious puberty. Hormonal studies established diagnosis of 11beta OHD: elevated plasma levels of 11-deoxycortisol, 17-hydroxyprogesterone, androstenedione and testosterone, low levels of cortisol and aldosterone, and suppressed plasma renin activity. Sequencing of the CYP11B1 gene identified compound heterozygous mutation consisting of a novel splicing mutation in intron 7 (IVS 7DS+4A to G) and R448H mutation in exon 8 previously reported mostly in Moroccan Jews. This is the first patient with CAH due to 11beta-OHD in Croatia (and Slavic population in general) in whom molecular diagnosis of CYP11B1 gene was performed. PMID- 20024694 TI - Antifreeze protein in Antarctic marine diatom, Chaetoceros neogracile. AB - The antifreeze protein gene (Cn-AFP) from the Antarctic marine diatom, Chaetoceros neogracile was cloned and characterized. The full-length Cn-AFP cDNA contained an open reading frame of 849 bp and the deduced 282 amino acid peptide chain encodes a 29.2 kDa protein, which includes a signal peptide of 30 amino acids at the N terminus. Both the Cn-AFP coding region with and without the signal sequence were cloned and expressed in Escherichia coli. Recombinant Cn AFPs were shown to display antifreeze activities based on measuring the thermal hysteresis and modified morphology of single ice crystals. Recombinant mature Cn AFP showed 16-fold higher thermal hysteresis activity than that of pre-mature Cn AFP at the same concentration. The ice crystal shape changed to an elongated hexagonal shape in the presence of the recombinant mature Cn-AFP, while single ice crystal showed a circular disk shape in absence of Cn-AFP. Northern analysis demonstrated a dramatic accumulation of Cn-AFP transcripts when the cells were subjected to freezing stress. This rapid response to freeze stress, and the antifreeze activity of recombinant Cn-AFPs, indicates that Cn-AFP plays an important role in low temperature adaptation. PMID- 20024695 TI - Changes in corticospinal excitability in the reactions of forearm muscles in humans to vibration. AB - Transcranial magnetic stimulation was used to study changes in corticospinal excitability during vibration of the flexor and extensor muscles of the wrist in healthy humans. The ratios of muscle stimulation responses to activity levels in these muscles on contraction associated with vibration (the tonic vibratory reflex, TVR) and after vibration of antagonist muscles in isometric conditions (the antagonist vibratory reflex, AVR) were analyzed. The normalized muscle response in the wrist flexors was found to increase by 66% compared with threshold values in the direct vibratory response (TVR), by 75% in the relayed vibratory response (AVR), and by 18% on voluntary contraction. However, increases in the motor response in vibratory responses as compared with those on voluntary contraction did not reach significance, which contrasted with the responses in the forearm flexors. These results are discussed from the point of view that the motor cortex plays different roles in vibratory responses in the distal and proximal muscles. PMID- 20024696 TI - Multiple sex partners and perceived risk of HIV infection in Zambia: attitudinal determinants and gender differences. AB - While there is much attention on perceived risk of HIV and subsequent cautionary behavior, evidence of the reciprocity of the relationship between recent risky sexual behavior and perceived risks of HIV infection remains scarce. This paper tests the hypothesis that having multiple sex partners within the last 12 months influences individual's perceived risk of HIV among Zambian men and women. It also examines attitudinal factors associated with having multiple sex partners and perceived HIV risk. Data come from the 2005 Household and Community Surveys of the Health Communication Partnership project. The sample includes 2610 men and women, and 445 community leaders in 36 districts. Tests of exogeneity confirm that having multiple sex partners is exogenous to perceived risk of HIV infection. Structural equation modeling is employed to test the hypothesis. Findings show that having multiple partners is the strongest predictor of perceived risk of HIV. Men were also more likely than women to have multiple sex partners, but less likely to consider themselves at risk of HIV, compared to women. Men were also more likely to be influenced by their peers and community members. Women were more likely than men to report some risk of HIV if they were aware of their partner's serostatus and had more interpersonal communication on HIV. Married women were less likely to consider themselves at risk than unmarried women. The findings indicate that it is essential for men to recognize the connections between unsafe sex behaviors and risks of HIV infection and that distinct intervention approaches to Zambian men and women are needed. While couple communication needs to be enhanced for both, community-based interventions that approach men are particularly important. PMID- 20024697 TI - We never thought this would happen: transitioning care of adolescents with perinatally acquired HIV infection from pediatrics to internal medicine. AB - PURPOSE: Transitioning the medical care of children with perinatally acquired HIV from pediatric care to internal medicine practices has become increasingly important as newer therapies prolong survival. The study aims to describe challenges to caring for these adolescents and the potential barriers to transitioning them to internal medicine-based care. METHODS: Qualitative study in which data were gathered from open-ended interviews conducted from November 2005 to April 2006 with 18 adolescents with HIV, 15 of their parents, and 9 pediatric health care providers from the Yale Pediatric AIDS Care Program, New Haven, Connecticut. RESULTS: Issues of stigma played a prominent role in both the challenges to care and barriers to transitioning care. Challenges to care were: (1) poor adherence to medication regimens; (2) adolescent sexuality; and (3) disorganized social environments. Potential barriers to transitioning care were: (1) families' negative perceptions of and experiences with stigma of HIV disease which undermined the desire to meet new providers; (2) perceived and actual lack of autonomy - pediatric providers feared that staff in adult clinics would demand a level of independence that adolescents did not have; and (3) difficulty letting go of relationships - adolescents, guardians, and providers described a familial relationship and expressed anxiety about terminating their relationships. CONCLUSION: Understanding these challenges and barriers can inform both pediatric and adult HIV care providers and enable them to create successful transition programs, with the goal of improving retention and follow-up to care. PMID- 20024698 TI - The "work" of women when considering and using interventions to reduce mother-to child transmission (MTCT) of HIV. AB - This paper explores HIV-infected women's experiences of considering and using recommended interventions during pregnancy and postpartum to reduce mother-to child transmission of HIV. Data were collected from 45 HIV-infected women aged 18 44 years living in Melbourne, Australia. A semi-structured interview was used to collect qualitative information on women's reproductive experience and intentions. The 15 women who had their children after their HIV diagnosis engaged in significant work including surveillance and safety work to minimise stigma and infection, information work to inform decisions and actions, accounting work to calculate risk and benefit, hope and worry work concerning a child's infection status and impact of interventions, work to redefine an acceptable maternal identity, work to prepare an alternative story to counter the disclosure effect of the intervention and emotional work to reconcile guilt when considering these interventions. This study provides a framework to help clinicians understand the real and on-going "work" that women engage in when they are considering interventions recommended by their physicians to reduce transmission of HIV. Even in circumstances where access to and acceptance of interventions are high, women continue to engage in this work even after they have a made a decision about a particular intervention. PMID- 20024699 TI - Availability of HIV-related health services in adolescent substance abuse treatment programs. AB - Given that alcohol and drug abuse heightens the risk of adolescents acquiring HIV, substance abuse treatment programs for youths may represent an important site of HIV prevention. In this research, we explored the adoption of three HIV related health services: risk assessment during intake, HIV prevention programing, and HIV testing. Data were collection through telephone interviews with 149 managers of adolescent-only substance abuse treatment programs in the USA. About half of these programs had adopted HIV risk assessment and HIV prevention. On-site HIV testing was less widely adopted, with only one in four programs offering this service. At the bivariate level, the availability of on site primary medical care and the availability of an overnight level of care were positively associated with these three types of services. The association for the measure of an overnight level of care was no longer significant once medical services were controlled. However, in a separate analysis, it was found that programs offering an overnight level of care were much more likely to offer on site medical care than outpatient-only facilities. There was also evidence that publicly funded treatment programs were more likely to offer HIV prevention and on-site HIV testing, after controlling for other organizational characteristics. Much more research about the adoption of HIV-related services in adolescent substance abuse treatment is needed, particularly to offer greater insight into why certain types of organizations are more likely to adopt these health services. PMID- 20024700 TI - Long-term efficacy and safety of polyalkylimide gel for the treatment of HIV associated lipoatrophy. AB - The long-term safety and efficacy of products used in the correction of HIV associated facial lipoatrophy (FLA) are largely unknown. The purpose of this study was to describe the long-term efficacy and safety of polyalkylimide gel (PAIG) in the treatment of HIV-associated FLA. In this open-label, randomized, single-center study, 31 HIV-positive individuals (median age 48 years (interquartile ranges (IQR) 45, 55, 97% male) with FLA were randomized to immediate (week 0 and six) or delayed (week 12 and 18) PAIG injections. Week 96 endpoints included change in FLA severity scores (FLSS) (five-point scale), proportion of patients with adverse events, and changes in quality of life, depression and anxiety using validated surveys. Results at week 96 were available for 28 patients. Adverse events, including swelling, redness, bruising and pain, were mild, and resolved after a median of three days following the injection. At week 96, median changes in physician and patient FLSS scores were -2 (IQR -3, -1; p<0.001 vs. baseline) and -2 (IQR -2, -1; p<0.001 vs. baseline), respectively. Physician and patient FLSS scores were not significantly different between the groups at week 96. Significant improvements in patient's anxiety (p<0.001), depression (p<0.001) and mental health (p=0.01) were observed from baseline to week 96. In conclusion, treatment with PAIG was associated with sustained improvements in both the physical and psychological components of FLA through 96 weeks of follow-up. PMID- 20024701 TI - Social networks and concurrent sexual relationships--a qualitative study among men in an urban South African community. AB - The aim was to explore and describe characteristics of males' social and sexual networks in a South African peri-urban community. Twenty in-depth interviews were conducted with men participating in a larger quantitative study where the median age of the men was 28.7 years and almost 56% had some high-school education, 17.2% were unemployed and 94.7% were not married. A Thematic Question Guide with open-ended questions was used for the interviews. A thematic content analysis was conducted to explore the characteristics and dynamics of social and sexual relationships among these men. A high number of temporary and stable concurrent female sexual partners, geographic mobility and high levels of unprotected sex were common. Increased status as a man and lack of trust in women's fidelity were given as important reasons for concurrent female sexual relationships. Strong social networks within male core groups provided economic and social support for the pursuit and maintenance of this behaviour. Concurrent sexual relationships in combination with high viral loads among newly infected individuals unaware of their HIV status create an extremely high-risk environment for the spread of HIV in this population. Interventions targeting men at high risk of HIV need to challenge current societal norms of masculinity to help promote individual sexual risk reduction strategies. Such strategies should go beyond increasing condom use, to include a reduction in the number of concurrent sexual partners. PMID- 20024702 TI - Development and validation of a culturally appropriate HIV/AIDS Stigma Scale for Puerto Rican health professionals in training. AB - HIV/AIDS stigma continues to be an obstacle for primary and secondary HIV prevention. Its consequences for the lives of people living with the disease have been well documented and continue to be of great concern for health care providers and researchers in the field. These consequences are worsened when such stigma emanates from health professionals, as this can limit access to services. One of the main obstacles for HIV/AIDS stigma research in Puerto Rico is the absence of quantitative measures to assess HIV/AIDS stigma manifestations among health professionals. In light of this gap in the scientific literature, the main objective of this study was to develop and test the psychometric properties of a culturally appropriate HIV/AIDS Stigma Scale for Puerto Rican health care providers and to develop a reduced form of the scale suitable for use in time limited clinical settings. The developed measure was based on previous qualitative evidence gathered from Puerto Rican health professionals (Varas-Diaz, Serrano-Garcia, & Toro-Alfonso, 2005) and administered to a sample of 421 health professionals in training. The scale addresses 12 HIV/AIDS stigma dimensions. In quantitative analyses 11 of these dimensions demonstrated satisfactory validity and reliability. These dimensions in turn were subcomponents of a higher-order general stigma factor. Implications and limitations of these findings are discussed. PMID- 20024703 TI - Validity and reliability of the Malay version of WHOQOL-HIV BREF in patients with HIV infection. AB - This study determines the validity and reliability of the Malay version of the World Health Organization Quality of Life (WHOQOL) assessment instrument in patients with human immunodeficiency virus (HIV) infection. A cross-sectional study on 157 patients with HIV seen at the Infectious Disease Unit, Hospital Raja Perempuan Zainab II, Kota Bharu, Kelantan was conducted. Factor analysis identified five major domains: physical needs, spirituality, social relationship, psychological, and environment. Significant correlation was found between each domain scores and the general health questions. The instrument was able to discriminate between asymptomatic and symptomatic HIV positive patients for all domain scores except for the spirituality domain. The internal consistency of the five domains ranged from 0.70 to 0.83. The intraclass correlation coefficient (ICC) ranged from 0.60 to 0.87 across all domains. In conclusion, the Malay version of WHOQOL-HIV BREF is a valid and reliable instrument in assessing quality of life in HIV positive patients. PMID- 20024704 TI - A strategy for selecting sexual partners believed to pose little/no risks for HIV: serosorting and its implications for HIV transmission. AB - A common HIV/AIDS risk reduction strategy among men who have sex with men (MSM) is to limit their unprotected sex partners to those who are of the same HIV status, a practice referred to as serosorting. Decisions to serosort for HIV risk reduction are based on personal impressions and beliefs, and there is limited guidance offered on this community derived strategy from public health services. This paper reviews research on serosorting for HIV risk reduction and offers an evidence-based approach to serosorting guidance. Following a comprehensive electronic and manual literature search, we reviewed 51 studies relating to the implications of serosorting. Studies showed that HIV negative MSM who select partners based on HIV status are inadvertently placing themselves at risk for HIV. Infrequent HIV testing, lack of HIV status disclosure, co-occurring sexually transmitted infections, and acute HIV infection impede the potential protective benefits of serosorting. Public health messages should continue to encourage reductions in numbers of sexual partners and increases in condom use. Risk reduction messages should also highlight the limitations of relying on one's own and partner's HIV status in making sexual risk decisions. PMID- 20024705 TI - More than ancillary: HIV social services, intermediate outcomes and quality of life. AB - In HIV care, the use of social or "ancillary" services to stabilize life situations and remove barriers to care is often seen as a means to the end of ensuring more consistent participation in medical care. By examining the impact of HIV social services on the achievement of intermediate outcomes (i.e., ceasing substance use, initiating anti-retroviral therapy (ART), and entering stable housing) and the relationship between intermediate outcome status and quality of life (QOL), our analysis aims to demonstrate the importance of achieving intermediate outcomes in and of themselves and thereby the importance of the ancillary services that assist clients in attaining desired intermediate outcomes. Our analysis relies on baseline and follow-up data from 1646 HIV positive participants collected during a longitudinal outcome evaluation of 23 HIV social service programs in the New York metropolitan area. Multivariate linear regression modeling was used to assess the impact of achieving intermediate outcomes on QOL at follow-up, controlling for baseline QOL, and demographic factors. The greatest improvements in QOL were found in individuals who changed their intermediate outcome status from using drugs to not using, from not using ART to using ART, and from being unstably housed to being stably housed. Our analysis strongly suggests the importance of achieving intermediate outcomes in improving QOL, and thereby the importance of social services that facilitate the achievement of these intermediate outcomes. The analysis also provides further validation of a QOL measure, by showing that it varies in systematic and expected ways with the achievement of intermediate outcomes. Our study suggests that social services are not merely ancillary in HIV care but rather crucial for achieving both intermediate outcomes as well as the final outcome of improved QOL. PMID- 20024706 TI - The effect of post-traumatic stress disorder on HIV disease progression following hurricane Katrina. AB - Post-traumatic stress disorder (PTSD) is a common psychological outcome of any disaster. The purpose of this study was to examine the effects of PTSD on disease progression among HIV-infected persons in metropolitan New Orleans post-hurricane Katrina. One-year post-storm, a convenience sample of 145 HIV-infected patients who returned to care at the HIV Outpatient Program clinic in New Orleans were interviewed. Clinical factors pre and one and two years post-disaster were abstracted from medical records and compared by PTSD status. Of the 145 participants, 37.2% had PTSD. Those with PTSD were more likely than those without PTSD to have detectable plasma viral loads at both follow-up time points post disaster and more likely to have CD4 cell counts <200/mm(3) two years post disaster. They were also more likely to have had medication interruptions immediately post-disaster. Our findings corroborate the findings of others that PTSD accelerates HIV disease progression. Disaster planners should consider the special counseling and medication safeguards needs of HIV-infected persons. PMID- 20024707 TI - Older African Americans' management of HIV/AIDS stigma. AB - The purpose of this study was to describe HIV/AIDS-related stigma in older African Americans living in the South, a population disproportionately affected by HIV/AIDS. Four focus groups were conducted with 24 men and women over 50 years old and a confirmed diagnosis of HIV. The focus group discussions were audiotaped and transcribed for analysis. Additionally, two stigma instruments, Self Perceptions of HIV Stigma, and Stigma Impact of HIV, were used to enhance the qualitative data from the focus groups. Constant comparative data analysis of the focus group discussions resulted in four themes related to HIV/AIDS stigma: (1) disclosure; (2) stigma experiences; (3) need for HIV/AIDS education; and (4) acceptance of the disease. Strategies to prevent or decrease anticipated stigma were described, such as selective or non-disclosure and not receiving care where they lived. The stigma instruments indicated that the participants had experienced the most stigma related to their internalized shame about having HIV disease, and had experienced little or no direct stigma. The study findings have implications for designing prevention programs, and strategies to improve social support for this age group. PMID- 20024708 TI - Barriers and facilitators to HIV testing and linkage to primary care: narratives of people with advanced HIV in the Southeast. AB - Persons with unrecognized HIV infection forgo timely clinical intervention and may unknowingly transmit HIV to partners. However, in the USA, unrecognized infection and late diagnosis are common. To understand barriers and facilitators to HIV testing and care, we conducted a qualitative study of 24 HIV infected persons attending a Southeastern HIV clinic who presented with clinically advanced illness. The primary barrier to HIV testing prior to diagnosis was perception of risk; consequently, most participants were diagnosed after the onset of clinical symptoms. While most patients were anxious to initiate care rapidly after diagnosis, some felt frustrated by the passive process of connecting to specialty care. The first visit with an HIV care provider was identified as critical in the coping process for many patients. Implications for the implementation of Centers for Disease Control and Prevention HIV routine screening guidelines are discussed. PMID- 20024709 TI - Common elements in self-management of HIV and other chronic illnesses: an integrative framework. AB - HIV/AIDS is widely recognized as a chronic illness within HIV care, but is often excluded from chronic disease lists outside the field. Similar to other chronic diseases, HIV requires lifetime changes in physical health, psychological functioning, social relations, and adoption of disease-specific regimens. The shift from acute to chronic illness requires a self-management model in which patients assume an active and informed role in healthcare decision making to change behaviors and social relations to optimize health and proactively address predictable challenges of chronic diseases generally and HIV specifically. This article reviews literature on chronic disease self-management to identify factors common across chronic diseases, highlight HIV-specific challenges, and review recent developments in self-management interventions for people living with HIV (PLH) and other chronic diseases. An integrated framework of common elements or tasks in chronic disease self-management is presented that outlines 14 elements in three broad categories: physical health; psychological functioning; and social relationships. Common elements for physical health include: a framework for understanding illness and wellness; health promoting behaviors; treatment adherence; self-monitoring of physical status; accessing appropriate treatment and services; and preventing transmission. Elements related to psychological functioning include: self-efficacy and empowerment; cognitive skills; reducing negative emotional states; and managing identity shifts. Social relationship elements include: collaborative relationships with healthcare providers; social support; disclosure and stigma management; and positive social and family relationships. There is a global need to scale up chronic disease self-management services, including for HIV, but there are significant challenges related to healthcare system and provider capacities, and stigma is a significant barrier to HIV-identified service utilization. Recognizing that self-management of HIV has more in common with all chronic diseases than differences suggests that the design and delivery of HIV support services can be incorporated into combined or integrated prevention and wellness services. PMID- 20024710 TI - Economic and quality of life outcomes of antiretroviral therapy for HIV/AIDS in developing countries: a systematic literature review. AB - The impacts of antiretroviral therapy (ART) on quality of life, mental health, labor productivity, and economic wellbeing for people living with HIV/AIDS in developing countries are only beginning to be measured. We conducted a systematic literature review to analyze the effect of ART on these economic and quality of life indicators in developing countries and assess the state of research on these topics. We searched Ovid/Medline, PubMed, Psych Info, Web of Science, Google Scholar, and the abstract database of the International AIDS Society Conference and the Conference on Retroviruses and Opportunistic Infections. Both qualitative and quantitative studies were included, as were peer-reviewed articles, gray literature, and conference abstracts and presentations. Findings are reported from 21 publications, including 14 full-length articles, six abstracts, and one presentation (representing 16 studies). Compared to HIV-positive patients not yet on treatment, patients on ART reported significant improvements in physical, emotional and mental health, and daily function. Work performance improved and absenteeism decreased, with the most dramatic changes occurring in the first three months of treatment and then leveling off. Little research has been done on the impact of ART on household wellbeing, with modest changes in child and family wellbeing within households where adults are receiving ART reportrd so far. Most studies from developing countries have not yet assessed economic and quality of life outcomes of therapy beyond the first year; therefore, longitudinal outcomes are still unknown. Findings were limited geographically, with an emphasis on sub Saharan Africa and adult treatment. As ART roll out extends throughout high HIV prevalence, low-resource countries and is sustained over years and decades, research on pediatric and differential gender economic and quality of life outcomes will become increasingly urgent, as will systematic evaluation of ART programs. PMID- 20024711 TI - The impact of taking or not taking ARVs on HIV stigma as reported by persons living with HIV infection in five African countries. AB - AIM: This study examined the impact of taking or not taking antiretroviral (ARV) medications on stigma, as reported by people living with HIV infection in five African countries. DESIGN: A two group (taking or not taking ARVs) by three (time) repeated measures analysis of variance examined change in reported stigma in a cohort sample of 1454 persons living with HIV infection in Lesotho, Malawi, South Africa, Swaziland, and Tanzania. Participants self-reported taking ARV medications and completed a standardized stigma scale validated in the African context. Data were collected at three points in time, from January 2006 to March 2007. Participants taking ARV medications self-reported a mean CD4 count of 273 and those not taking ARVs self-reported a mean CD4 count of 418. RESULTS: Both groups reported significant decreases in total HIV stigma over time; however, people taking ARVs reported significantly higher stigma at Time 3 compared to those not taking ARVs. DISCUSSION: This study documents that this sample of 1454 HIV infected persons in five countries in Africa reported significantly less HIV stigma over time. In addition, those participants taking ARV medications experienced significantly higher HIV stigma over time compared to those not taking ARVs. This finding contradicts some authors' opinions that when clients enroll in ARV medication treatment it signifies that they are experiencing less stigma. This work provides caution to health care providers to alert clients new to ARV treatment that they may experience more stigma from their families and communities when they learn they are taking ARV medications. PMID- 20024712 TI - Experience of sexual violence among women in HIV discordant unions after voluntary HIV counselling and testing: a qualitative critical incident study in Uganda. AB - HIV-serodiscordant relationships are those in which one partner is infected with HIV while the other is not. We investigated experiences of sexual violence among women in HIV discordant unions attending HIV post-test club services in Uganda. A volunteer sample of 26 women from three AIDS Information Centres in Uganda who reported having experienced sexual violence in a larger epidemiological study were interviewed, using the qualitative critical incident technique. Data were analysed using TEXTPACK, a software application for computer-assisted content analysis. Incidents of sexual violence narrated by the women included use of physical force and verbal threats. Overall, four themes that characterise the women's experience of sexual violence emerged from the analysis: knowledge of HIV test results, prevalence of sexual violence, vulnerability and proprietary views and reactions to sexual violence. Alcohol abuse by the male partners was an important factor in the experience of sexual violence among the women. Their experiences evoked different reactions and feelings, including concern over the need to have children, fear of infection, desire to separate from their spouses/partners, helplessness, anger and suicidal tendencies. HIV counselling and testing centres should be supported with the capacity to address issues related to sexual violence for couples who are HIV discordant. PMID- 20024713 TI - Expanding the lens of HIV services provision in Canada: results of a national survey of HIV health professionals. AB - Those living with HIV may experience a range of disabilities, including body impairments, activity limitations, and social participation restrictions. The aim of this study was to examine HIV services provision in Canada by exploring practices, referrals, and service delivery challenges from the perspective of HIV health professionals (including nurses, physicians, social workers, pharmacists, psychologists, and dieticians), and to explore differences in referrals and perceived service delivery challenges by professional group, jurisdiction, community size, and practice in a Northern region. We conducted a nationwide mail survey with the population of selected HIV health professionals in Canada using the Dillman tailored design survey method. Of the 731 deliverable mailings, we received 462 (63%) responses, with 36% of eligible respondents completing the survey (n=214). The large majority (90%) of HIV professionals were located in metropolitan or urban communities and worked predominantly in hospital in-patient (42%), out-patient (50%), and HIV specialty clinic (46%) settings in one of the three provinces (Ontario, Quebec, and British Columbia) with the highest HIV prevalence. HIV health professionals referred primarily, and at relatively high levels, to AIDS service organizations (79%) and social workers (84%) to address participation restrictions and social issues; a lower percentage referred to rehabilitation professionals and other service providers to address impairments, activity limitations, or participation restrictions. Of respondents, 74% perceived barriers to care specific to HIV. Our results suggest that there is little difference in referral patterns by profession, jurisdiction, community size, or northern region of practice. There is a need for increased information and education of HIV health professionals that may refer to rehabilitation and other health services. In addition, new approaches are needed to coordinate multisectoral care and enhance the access and delivery of HIV rehabilitation health services to better meet the disablement needs of people living with HIV in Canada. PMID- 20024714 TI - Voices on adherence to ART in Ethiopia and Uganda: a matter of choice or simply not an option? AB - This paper explores HIV patients' adherence to antiretroviral treatment (ART) in resource-limited contexts in Uganda and Ethiopia, where ART is provided free of charge. Qualitative semi-structured interviews were conducted with 79 patients, 17 peer counselors, and 22 providers in ART facilities in urban and rural areas of Ethiopia and Uganda. Interviewees voiced their experiences of, and views on ART adherence both from an individual and a system level perspective. Two main themes emerged from the content analysis: "Patients' competing costs and systems' resource constraints" and "Patients' trust in ART and quality of the patient provider encounters." The first theme refers to how patients' adherence was challenged by difficulties in supporting themselves and their families, paying for transportation, for drug refill and follow-up as well as paying for registration fees, opportunistic infection treatment, and expensive referrals to other hospitals. The second theme describes factors that influenced patients' capacity to adhere: personal responsibility in treatment, trust in the effects of antiretroviral drugs, and trust in the quality of counseling. To grant patients a fair choice to successfully adhere to ART, transport costs to ART facilities need to be reduced. This implies providing patients with drugs for longer periods of time and arranging for better laboratory services, thus not necessitating frequent revisits. Services ought to be brought closer to patients and peripheral, community-based healthworkers used for drug distribution. There is a need for training providers and peer counselors, in communication skills and adherence counseling. PMID- 20024715 TI - The cost effectiveness of integrated care for people living with HIV including antiretroviral treatment in a primary health care centre in Bujumbura, Burundi. AB - The incremental cost effectiveness of an integrated care package (i.e., medical care including antiretroviral therapy (ART) and other services such as psychological and social support) for people living with HIV/AIDS was calculated in a not-for-profit primary health care centre in Bujumbura run by Society of Women against AIDS-Burundi (SWAA-Burundi), an African non-governmental organisation (NGO). Results are expressed as cost-effectiveness ratio 2007, constant US$ per disability-adjusted life year (DALY) averted. Unit costs are estimated from the NGO's accounting data and activity reports, healthcare utilisation is estimated from the medical records of a cohort of 149 patients. Effectiveness is modelled on the survival of this cohort, using standard calculation methods. The incremental cost of integrated care for people living with HIV/AIDS in the Bujumbura health centre of SWAA-Burundi is 258 USD per DALY averted. The package of care provided by SWAA-Burundi is therefore a very cost effective intervention in comparison with other interventions against HIV/AIDS that include ART. It is however, less cost effective than other types of interventions against HIV/AIDS, such as preventive activities. PMID- 20024716 TI - Sexual risk behavior among early initiators of sexual intercourse. AB - The aim of this study was to examine sexual risk behavior among adolescents in the United States classified as early initiators of sexual intercourse. Secondary analyses were carried out with data from the 2007 Youth Risk Behavior Survey (n=14,041). Only adolescents 16-18 years old who reported being sexually experienced were selected for analyses (n=5315, 48% female). Of sexually experienced adolescents, 11% were classified as early initiators of sexual intercourse. Based on prior research it was expected that early initiators of sexual intercourse would demonstrate increased sexual risk behavior when compared to adolescents who initiated sexual intercourse at later ages and that there would be ethnic and gender differences in risk behavior. Early initiators were significantly more likely to report multiple sex partners in their lifetime and were more likely to drink or use drugs before the last time they had sex. Overall, the findings did not demonstrate differences in sexual risk based on gender and ethnicity. PMID- 20024717 TI - State of the ART: clinical efficacy and improved quality of life in the public antiretroviral therapy program, Free State province, South Africa. AB - The South African public-sector antiretroviral treatment (ART) program has yielded promising early results. To extend and reinforce these preliminary findings, we undertook a detailed assessment of the clinical efficacy and outcomes over two years of ART. The primary objective was to assess the clinical outcomes and adverse effects of two years of ART, while identifying the possible effects of baseline health and patient characteristics. A secondary objective was to address the interplay between positive and negative outcomes (clinical benefits versus adverse effects) in terms of the patients' physical and emotional quality of life (QoL). Clinical outcome, baseline characteristics, health status, and physical and emotional QoL scores were determined from clinical files and interviews with 268 patients enrolled in the Free State ART program at three time points (6, 12, and 24 months of ART). Age, sex, education, and baseline health (CD4 cell count and viral load) were all independently associated with the ART outcome in the early stages of treatment, but their impact diminished as the treatment progressed. The number of patients classified as treatment successes increased over the first two years of ART, whereas the proportion of patients experiencing adverse effects diminished. Importantly, our findings show that ART had strong and stable positive effects on physical and emotional QoL. These favorable results demonstrate that a well-managed public-sector ART program can be very successful within a high-HIV-prevalence resource-limited setting. This finding emphasizes the need to adopt treatment scale-up as a key policy priority, while at the same time ensuring that the highest standards of healthcare provision are maintained. Healthcare services should also target vulnerable groups (males, less-educated patients, those with low baseline CD4 cell counts, and high baseline viral loads) who are most likely to experience treatment failure. PMID- 20024718 TI - Treatment partners and adherence to HAART in Central Mozambique. AB - Adherence to highly active antiretroviral treatment (HAART) has been associated with increased survival rates and decreased drug resistance in various settings. There is growing concern that loss to follow-up will increase and adherence rates will decrease as HAART programs are expanded in resource-limited settings. In Central Mozambique, an innovative program was implemented, using community-based (trained community activists) and self-selected (family members or friends) "treatment partners" to provide psycho-social support to patients on HAART. We calculated adherence rates based on pharmacy records for all patients who refilled their medication for at least six consecutive months between September 2004 and June 2006. Medical charts were reviewed for a subset of 375 patients having high (> or =90%) adherence and 59 patients having low (<90%) adherence. Multivariate logistic regression analysis assessed the association between the type of treatment partner used and adherence to HAART. A total of 305 patients (70%) had self-selected treatment partners, 121 (28%) had community-based treatment partners, and 8 (2%) had no treatment partner. In adjusted analysis, patients who had no treatment partner were more likely to have low adherence (OR 9.47; 95% confidence interval 2.37-37.86 compared to self-selected treatment partner). Patients with community-based treatment partners did not have significantly lower adherence than patients with self-selected treatment partners. While it cannot be determined from these data which aspects or types of peer support are most effective in maintaining adherence, it appears that peer support was beneficial to this study population. While the study results are not directly applicable to other populations, other HAART programs should consider the potential benefit of providing treatment support to patients. PMID- 20024719 TI - The risk of HIV and HCV infections among injection drug users in northeast India. AB - Injection drug users (IDUs) and their associated risk behavior are responsible for driving the human immunodeficiency virus (HIV) epidemic in northeast India. So a group of IDUs from two northeastern states (Mizoram and Nagaland) of India were studied to find the prevalence of HIV, co-infection with hepatitis C virus (HCV), hepatitis B virus (HBV), and associated risk behaviors. Out of the 400 IDUs enrolled, 398 consented for HIV, HCV, and hepatitis B surface antigen (HbsAg) test. Of them, 10.8% were HIV-1 antibody positive, 47.8% had HCV antibody, and 3.8% had detectable HBsAg. Among the HIV infected subjects, 79.1% were co-infected with HCV and 6.9% had triple infection. Heroin users showed a higher association with HIV (OR = 7.3, 95% CI: 2.5-21.5, p=0.0003) and HCV infection (OR = 7.6, 95% CI: 3.5-16.6, p<0.0001) than Spasmo-proxyvon (dextropropoxyphene, a synthetic opiod analgesic). In summary, apart from the known risk variables among IDUs, type of injecting drugs also influences the HIV/HCV transmission pattern among the IDUs. PMID- 20024720 TI - Self-fulfillment despite barriers: volunteer work of people living with HIV. AB - With the decline in the morbidity and mortality associated with HIV infection, the majority of people living with HIV (PWHIV) experience a higher quality of life and longer life expectancy. Since this diagnosis no longer prevents an active lifestyle, many PWHIV are re-integrating into the workplace or are contemplating this possibility. Despite the multiple advantages associated with a return to work, research has identified barriers related to work and HIV infection. These barriers could prevent an important minority of PWHIV who wish to return to work from re-integrating into this environment. In this context, volunteering could be an alternate way to regain an active lifestyle. This research found that volunteer work allowed participants to enrich their social lives, to regain a sense of psychological well-being, and to apply their abilities to the benefit of others. However, participants are restricted by their preference to volunteer for HIV-related organizations in order to avoid social stigma and rejection. PMID- 20024721 TI - Trends in hospitalizations with psychiatric diagnoses among HIV-infected women in the USA, 1994-2004. AB - Psychiatric illnesses commonly co-occur with HIV infection and such illnesses have been linked to women's poorer medication adherence and suicide. Using hospital discharge data from the 1994-2004 Nationwide Inpatient Sample, we conducted this study to describe hospitalizations with psychiatric diagnoses from 1994 through 2004 and evaluate the association of specific psychiatric disorders among hospitalized HIV-infected women in the USA with their lack of adherence to medical treatment and suicide attempt. Multivariable logistic regression analyses were used to examine trends in hospitalizations with psychiatric diagnoses among nonpregnant HIV-infected women and the association between specific disorders and women's lack of adherence to medical treatment and suicide attempt. Between 1994 and 2004, the estimated number of all hospitalizations among nonpregnant HIV infected women increased by 8%, while the number of hospitalizations with a psychiatric diagnosis in this population increased by 73%. After adjusting for demographic factors and alcohol/substance abuse, we found that HIV-infected women were more likely to be hospitalized for mood (odds ratio (OR): 2.35; 95% confidence interval (CI): 1.93-2.88), anxiety (OR: 2.24, 95%CI: 1.74-2.88), and psychotic (OR: 1.45, 95%CI: 1.10-1.90) disorders in 2004 than in 1994. There was a significant association of alcohol/substance abuse with mood, adjustment, anxiety, personality, and psychotic disorders. Noncompliance with medical treatment was significantly associated with psychotic disorders, whereas suicide attempt/self-inflicted injury was significantly associated with mood, adjustment, anxiety, personality, and psychotic disorders. The number of hospitalizations with a psychiatric diagnosis among HIV-infected women in the USA has increased substantially. As HIV-infected women live longer, these results highlight the need for targeted public health interventions to address mental health issues in this population. PMID- 20024722 TI - Female sex work and HIV risks in Croatia. AB - The aim of this paper was to analyze and compare the prevalence of HIV-related sexual risk taking among Croatian female sex workers (FSW) in two major urban areas. Two groups of FSWs were interviewed in Zagreb (n=65) and Split (n=89). Participants' mean age was 33.3 (SD=8.32). Interviews were conducted by outreach organizations that provide health services to sex workers in the two cities. The study used a brief questionnaire with standardized behavioral and HIV knowledge indicators. The two groups of FSWs differed significantly in most socio demographic and socio-sexual indicators. Women from the Split sample were somewhat younger (chi(2)=6.87, p<0.05), less educated (chi(2)=7.71, p<0.05), less likely to be single (chi(2)=19.81, p<0.001), and less likely to be unemployed (chi(2)=5.22, p<0.05). In addition, they injected drugs in higher proportion (chi(2)=35.03, p<0.001), but had less clients in the preceding month (chi(2)=12.54, p<0.001) and were less likely to be abused by them (chi(2)=7.18, p<0.01). HIV testing was significantly more prevalent among participants in the Split sample (chi(2)=4.95, p<0.05). In multivariate analysis, selling sex in Zagreb (OR = 14.48, p<0.01), having secondary or higher education (OR = 4.76, p<0.05), ever tested for HIV (OR = 8.34, p<0.05), and having assessed the risk of getting infected with HIV as high (OR = 0.23, p<0.05) were significantly associated with consistent condom use with clients in the last month. The findings of this first systematic study on HIV-related risks among FSWs in Croatia point to the need to update targeted intervention programs by improving the prevention of HIV risks associated with injecting drug use (Split) and by a more efficient HIV educational approach. PMID- 20024723 TI - Emotional and behavioral problems and mental health service utilization of youth living with HIV acquired perinatally or later in life. AB - This study sought to examine predictors of psychological symptoms and psychiatric service receipt among youth with HIV. Data were from the baseline assessment of Adolescent Impact, a study of 13-21-year-old youth with HIV in three US cities. Between August 2003 and February 2005, participants completed the age-appropriate youth or adult self-report symptom checklists (Achenbach system of empirically based assessment) and reported their psychiatric treatment history. Psychiatric diagnoses were abstracted from medical records. The 164 participating youth living with HIV were Black (81%), female (52%), Heterosexually identified (62%), and perinatally HIV-infected (60%). Thirty-one percentage reported levels of internalizing (i.e., self-focused/emotional), externalizing (i.e., outwardly focused/behavioral), or overall symptoms consistent with clinical psychopathology. In multivariate analyses, questioning one's sexual identity was associated with greater internalizing problems, whereas identifying as Bisexual was associated with greater externalizing problems (p<0.05). Symptoms were not associated with HIV transmission group. Participants with > or =1 composite score within the clinical range were more likely to have received > or =1 psychiatric service (Odds ratio (OR): 2.51; 95% confidence interval (CI): 1.22, 5.13) and a psychiatric diagnosis in the past year (OR: 2.16; 95% CI: 1.09, 4.27). However, 27% with clinically elevated scores had never received psychiatric care. Results suggest that among youth with HIV, those who identify as Bisexual or Questioning are at greatest risk for emotional and behavioral problems. Despite available mental health services, some youth with HIV are not receiving needed mental health care. Enhanced evaluation, referral and mental health service linkage is needed for these high-risk youth. PMID- 20024724 TI - Mediators of HIV-related stigma and risk behavior in HIV infected young women. AB - Stigma in HIV positive persons has been associated with numerous negative sequelae, including decreased social support, depressive symptoms, and engagement in risk behaviors. Few studies examined the interrelationships of these factors to facilitate understanding of the mechanisms by which HIV stigma influences risk behavior, thus the current study focuses on identifying pathways between HIV related stigma and risk behavior in 147 young HIV positive women. Depression and social support were hypothesized to mediate between HIV-related stigma and risk behavior. Structural equation modeling was used to test these hypothesized pathways, results suggested that depression was a significant mediator between HIV-related stigma and risk behavior. Implications for interventions with young HIV positive women who report high levels of HIV-related stigma include a focus on depression as a method of reducing engagement in risk behavior and improving mental health and health behaviors in persons living with HIV. PMID- 20024725 TI - Effectiveness of a training program to increase the capacity of health care providers to provide HIV/AIDS care and treatment in Swaziland. AB - Implementation of HIV care and treatment programs in sub-Saharan Africa is a complex undertaking that requires training of health care providers (HCPs). Many sub-Saharan African countries have introduced training programs to build human resources for health. Evaluation of the ongoing trainings is warranted so that programs can be improved. The purpose of this study was to evaluate Baylor International Pediatric AIDS Initiative's (BIPAI) HCP training program in Swaziland. The specific aims were: (1) to assess coverage and delivery of the training program; and (2) to determine the impact of the training program on HCPs' knowledge about HIV and pediatric practices, attitudes toward HIV/AIDS patients, and self-efficacy to provide antiretroviral therapy (ART). The evaluation was a multimethod design with two types of data collection and analysis: (1) one-group pretest-posttest survey with 101 HCPs; and (2) semi structured in-depth interviews with seven trainers from Baylor College of Medicine and 16 local HCPs in Swaziland. Quantitative data were analyzed using Stata Statistical Software version 8.2 for descriptive and multivariate analysis while factor analysis was done using Statistical Program for Social Sciences version 14. The transcribed interviews were analyzed using a didactic approach. Process evaluation showed that the training had good coverage, was delivered as intended, and improved as the work progressed. The training program led to a significant increase (p=0.0000) in HCPs' knowledge about HIV/AIDS, ART, and relevant clinical pediatrics practices between pretest (mean 68.7% SD 13.7) and post training (mean 84.0% SD 12.0). The training program also increased trainees' self-efficacy to provide ART and their attitudes toward AIDS patients (p=0.0000 and 0.02, respectively). In conclusion, BIPAI training program in Swaziland had good coverage of all health care facilities and HCPs in Swaziland. The training was effective in imparting knowledge and skills to HCPs and in their attitudes toward HIV/AIDS patients. PMID- 20024726 TI - Social-cognitive determinants of condom use in a cohort of young gay and bisexual men. AB - The aim of this prospective study was to identify relevant determinants of young gay and bisexual men's (YGBM) condom use when having anal sex with casual partners. Respondents (185 YGBM in the midst of their coming-out; mean age 18.9 years) completed an online questionnaire on social-cognitive determinants of condoms use derived from the theory of planned behavior (Ajzen, 1991) at Wave 1. At six months follow-up (Wave 2) sexual behavior with casual partners was assessed. A total of 63 YGBM reported sex with a casual partner in the six months between Waves 1 and 2, of whom 49% (N=31) had anal sex. Of the YGBM who had anal sex, 42% (N=13) had unprotected anal sex. Condom use with casual partners was best predicted by the intention to always use condoms. Furthermore, attitude, descriptive and personal norms, and perceived control significantly predicted intention to always use condoms. Interventions, targeting YGBM, aiming to promote condom use with casual partners should focus on increasing attitudes and strengthening skills to negotiate and use condoms. PMID- 20024727 TI - Stressful or traumatic life events, post-traumatic stress disorder (PTSD) symptoms, and HIV sexual risk taking among men who have sex with men. AB - The present study assessed the presence of post-traumatic stress disorder (PTSD) symptoms in response to stressful or traumatic life events and their impact on HIV risk behaviors and associated psychosocial variables among men who have sex with men (MSM). Participants (n=189; 60% HIV-infected) who were recruited by notices posted in a community health clinic and via a modified respondent-driven sampling technique completed a behavioral assessment survey. Sixty percentage of participants screened positive for having PTSD symptoms using the startle, physiological arousal, anger, and numbness screening instrument. After controlling for race, sexual self-identification, and HIV status, multivariable logistic regression analyses revealed that screening in for having PTSD symptoms was significantly associated with having engaged in unprotected anal (insertive or receptive) sex in the past 12 months, over and above any effects of whether or not a traumatic/stressful event occurred during the year (adjusted odds ratio [OR] = 2.72; p<0.02; 95% confidence interval [CI] = 1.19-6.20). In addition, MSM with PTSD symptoms were more likely to have clinically significant depressive symptoms (adjusted OR = 3.50; p<0.001) and/or symptoms of social anxiety (adjusted OR = 2.87; p<0.01; 95% CI = 1.48-5.62). The current study, in the context of other research documenting the high rates of co-occurring psychosocial issues facing MSM, points to the importance of incorporating coping with these issues in HIV and sexually transmitted disease prevention and care interventions. PMID- 20024728 TI - Psychological well-being and socio-economic hardship among AIDS orphans and other vulnerable children in Guinea. AB - Over the past decade, the effects of AIDS-related parental death on children's socio-economic, educational and psychological well-being have become apparent. Most studies, however, have compared the plight of so-called AIDS orphans with non-orphaned children only. Consequently, such study designs are unable to establish if the AIDS-related cause of death of the parents confers effects additional to those of parent-bereavement. We therefore conducted a cross sectional survey to assess the psychological well-being and socio-economic hardship among 140 non-orphaned children, 133 children orphaned by causes other than AIDS (O) and 124 children orphaned by AIDS (O-A) in Conakry, N'Zerekore and the villages around N'Zerekore, Guinea. Multi-way analysis of variance and multiple (ordinal) logistic regression models were applied to measure the association between the orphan status and psychological well-being, school attendance, economic activities, frequency of going to bed hungry and sleeping commodity. After adjustment for confounding factors, the psychological well-being score (PWS) was significantly lower among AIDS-orphaned children than among O (P<0.001). Additionally, AIDS-orphaned children were more likely to be engaged in economic activities (adjusted odds ratio (AOR) = 3.04; 95% CI: 1.45-6.36) and to go to bed hungry on a daily basis (AOR = 2.73; 95% CI: 1.24-6.02) than other orphans. The differences in school attendance and the proportion of children with a bed or couch to sleep between AIDS-orphaned children and O were not statistically significant. This situation calls for sustainable and holistic approaches to ensure the psychological and socio-economic stability of AIDS orphans and other vulnerable children. PMID- 20024729 TI - Effects of structural intervention on increasing condom availability and reducing risky sexual behaviours in gay bathhouse attendees. AB - The study purpose was to evaluate the effectiveness of a structural intervention in reducing unprotected sexual behaviours, increasing condom accessibility and their relationship to the prevalence of HIV infection and sexually transmitted infections (STIs) in gay bathhouse attendees. A quasi-experimental study with a non-equivalent control group for comparison was conducted at nine gay bathhouses in Taiwan from 2004 to 2006. A structural intervention designed to increase condom distribution and accessibility inside the bathhouses was implemented at one bathhouse in each city. Bathhouse attendees were invited to complete a questionnaire and to be screened for HIV infection and STIs. Of 632 study participants, 270 were surveyed six months after the intervention was conducted. At the time of follow-up after controlling for baseline data, patrons attending bathhouses for which intervention was performed were more likely to report accessing condoms inside bathhouses than those attending control bathhouses (92.6% versus 81.3%, P = 0.016), and condoms were more likely to be available at the reception desk of the bathhouse entrance (87.5% in intervention bathhouses versus 69.4% in control, P = 0.047). In a multivariate analysis adjusted for age, access to condoms inside bathhouses (adjusted odds ratio (AOR): 4.35; 95% confidence interval (CI): 1.48-12.78) and attendance at bathhouses with intervention (AOR: 2.07; 95% CI: 1.07-3.97) were independently associated with consistent condom use during anal intercourse. There were no significant differences in the prevalences of HIV infection and STIs at the six-month follow up. Bathhouses with structural intervention were associated with consistent condom use during anal intercourse among bathhouse patrons. Our findings highlight the importance of extension of condom distribution inside the bathhouses to increase condom use among bathhouse patrons. PMID- 20024730 TI - Positive future orientation as a mediator between traumatic events and mental health among children affected by HIV/AIDS in rural China. AB - OBJECTIVE: The current study was designed to explore the effect of future orientation in mediating the relationship between traumatic events and mental health in children affected by HIV/AIDS in rural China. METHODS: Cross-sectional data were collected from 1221 children affected by HIV/AIDS (755 AIDS orphans and 466 vulnerable children). Future orientation among children was measured using three indicators (future expectation, hopefulness toward the future, and perceived control over the future). Measures of mental health consisted of depression, loneliness, and self-esteem. Children's experience of any traumatic events was measured using a modified version of the Life Incidence of Traumatic Events-Student Form. Mediation analysis was conducted using structural equation modeling (SEM) methods. RESULTS: Among the children surveyed, most of the traumatic indicators were negatively associated with future expectation, hopefulness, perceived control, and self-esteem, and positively associated with depression and loneliness. The SEM of mediation analysis demonstrated an adequate fit. Future orientation fully mediated the relationship between traumatic events and mental health and accounted for 67.9% of the total effect of traumatic events on mental health. CONCLUSIONS: Results of this study support the positive effect of future expectation in mediating the relationship between traumatic events and mental health among children affected by HIV/AIDS in China. Future mental health promotion and intervention efforts targeting children affected by HIV/AIDS should include components that can mitigate the negative impact of traumatic events on their lives. These components may aim to develop children's positive future expectations, increase their hopefulness toward the future, and improve their perceived control over the future. PMID- 20024731 TI - Quality of life, psychosocial health, and antiretroviral therapy among HIV positive women in Zimbabwe. AB - Little is known about the psychosocial impact of antiretroviral therapy (ART) among women in sub-Saharan Africa. Therefore, we conducted a cross-sectional study in Zimbabwe to assess the impact of ART on HIV-positive women's health related quality of life, using the Medical Outcomes Study-HIV Quality of Life (QOL) questionnaire. Additionally, we assessed socio-demographics, reproductive and sexual health, HIV-related history, disclosure, social stigma, self-esteem, and depression. Structured interviews were conducted with 200 HIV-positive women and categorized into three groups by treatment: (1) Group 1 (n=31) did not meet clinical or laboratory criteria to begin treatment; (2) Group 2 (n=73) was eligible to begin treatment but awaiting initiation of treatment; and (3) Group 3 (n=96) was on ART for a median of 13 months. The women had similar socio demographic characteristics but varied significantly in clinical characteristics. Women on ART reported fewer AIDS-related symptoms in the last week and year and had higher current and lower baseline CD4 counts compared to women not on ART. On most QOL domains women on ART reported higher mean scores as compared to women not on ART (p<0.01). Additionally, women on ART reported less depression compared to women not on ART (p<0.001). Between the two groups of women not on ART, unexpectedly, there were no significant differences in their scores for QOL or depression. Thus, Zimbabwean women living with HIV experience better overall QOL and lower depression on ART. Altogether, our findings suggest that ART delivery in resource-poor communities can enhance overall QOL as well as psychosocial functioning, which has wide-ranging public health implications. PMID- 20024732 TI - Patients and family care givers' experiences around highly active antiretroviral therapy (HAART). AB - Even with advances in the medical management of HIV/AIDS, it remains an important cause of morbidity and mortality. Network members are a source of support and may be designated as the patient's surrogate therapeutic decision maker. However, little is known about the role of caregivers regarding highly active antiretroviral therapy (HAART) and adherence to medication, especially among low income and medically underserved HIV-infected individuals. The objective of this analysis was to explore patients and their caregivers' understanding, insight, and perceptions of HAART. This was a qualitative study conducted in an urban teaching hospital in the Bronx, NY, consisting of 144 adults with advanced HIV/AIDS and their informal/familial caregivers. Patients and caregivers completed in-depth interviews and brief questionnaires. The data were analyzed using standard qualitative techniques. Themes related to HAART efficacy, side effects, and adherence were identified in the discussions with both patients and their care providers. Looking for consistencies and discordant reports of patients-caregivers dyadic experience with HAART and their association with healthcare sentiment, it was found that the accounts of both members of the dyad were more likely to reflect positive feelings about the patient's healthcare experience when they shared perceptions about the treatment, and were more likely to have a negative viewpoint when the dyad was discordant about treatment. These findings support the importance for HIV healthcare providers to include both patients and their caregivers in discussions about HAART, in order to improve their understanding of and satisfaction with the medication, and, to ultimately contribute to patients' adherence. The study also suggests that healthcare teams may enhance dialog with patients and caregivers to create therapeutic decisions to accommodate the priorities and values of the patient and their family. Clinical, educational, and evaluating tools need to be further developed and evaluation to facilitate this process. PMID- 20024733 TI - Male involvement in antenatal HIV counseling and testing: exploring men's perceptions in rural Malawi. AB - Antenatal care can act as an excellent tool to improve access to HIV counseling and testing services. This paper investigates an issue that may weaken its potential, namely lack of male involvement. We explored married men's perceptions of HIV in pregnancy and male involvement in antenatal HIV testing and counseling in Southern Malawi through 11 focus group discussions and a cross-sectional survey (n=388). The main findings were that men were largely unaware of available antenatal HIV testing and counseling services, and perceived it overall problematic to attend female-oriented health care. Most men supported provision of antenatal HIV testing. They perceived husbands to participate in the process indirectly through spousal communication, being faithful during pregnancy, and supporting the wife if found HIV-positive. Involvement of husbands was compromised by men's reluctance to learn their HIV status and the threat that HIV poses on marriage. Men stressed the importance of prior spousal agreement of antenatal HIV testing and considered HIV testing without their consent a valid reason for divorce. We suggest that male involvement in antenatal HIV testing requires refocusing of information and health services to include men. To avoid negative social outcomes for women, comprehensive and early involvement of men is essential. PMID- 20024734 TI - Costs of care for people living with combined HIV/AIDS, chronic mental illness, and substance abuse disorders. AB - To determine healthcare access and costs for triply diagnosed adults, we examined baseline data from the HIV/AIDS Treatment Adherence, Health Outcomes and Cost Study, a multi-site cohort study of HIV+ adults with co-occurring mental and substance abuse disorders conducted between 2000 and 2004. Baseline interviews were conducted with 1138 triply diagnosed adults in eight predominantly urban sites nationwide. A modified version of Structured Interview for DSM-IV Axis I Disorders (SCID) was used to assign Diagnostic and Statistical Manual of Mental Disorders, 4th edition (DSM-IV) diagnoses for the preceding year. Utilization of a broad range of inpatient and outpatient services and medications over the preceding three months was patient-reported in face-to-face interviews. We then applied nationally representative unit costs to impute average monthly expenditures. We measured (poor) access to care during the three-month period by whether the patient had: (a) no outpatient medical visits; (b) at least one emergency room visit without an associated hospitalization; and (c) at least one hospitalization. At baseline, mean expenditures were $3880 per patient per month. This is nearly twice as high as expenditures for HIV/AIDS patients in general. Inpatient care (36%), medications (33%), and outpatient services (31%) each accounted for roughly one-third of expenditures. Expenditures varied by a factor of 2:1 among subgroups of patients, with those on Medicare or Medicaid, not in stable residences, or with poor physical health or high viral loads exhibiting the highest costs. Access to care was worse for women and those with low incomes, unstable residences, same-sex exposure, poor physical or mental health, and high viral loads. We conclude that HIV triply diagnosed adults account for roughly one fifth of medical spending on HIV patients and that there are large variations in utilization/costs across patient subgroups. Realized access is good for many triply diagnosed patients, but remains suboptimal overall. Deficiencies in HIV care are unevenly distributed, tending to concentrate on already disadvantaged populations. PMID- 20024735 TI - The phenomenology of body image in men living with HIV. AB - Research into how people living with HIV or AIDS (PLWHA) experience and make sense of feared or actual body changes, such as lipodystrophy, is limited. The present study conducted in-depth interviews with gay men living with HIV. Interpretative phenomenological analysis (IPA) yielded themes across five domains. The ambiguity of early-stage lipodystrophy was a cause of distress. There was a drive to detect changes early, leading to anxiety, uncertainty and negative feelings about the body as well as possible misperception of change. In later stages, lipodystrophy was felt to be highly distinctive. Participants struggled to live with a shape that transgressed a body ideal. Feelings of loss of control were evident in both the increased ineffectiveness of strategies to maintain a desirable appearance and in the tendency for such changes to act as a visible marker of status. Conflicting feelings emerged in ideas of thinness and of health, with loss of fat seen as desirable in certain contexts. The study builds on previous research suggesting that the high value of appearance, particularly within gay communities, may lead to extreme compensatory behaviours. HIV places increased risk on a group highly vulnerable to body dissatisfaction and eating disorders. The study concludes with a cognitive-behavioural model of body image for PLWHA and suggestions for intervention. Further research is needed to validate the model and investigate whether the findings are generalisable. However, body image concerns should be acknowledged when addressing HIV-related health. PMID- 20024736 TI - Survival of an AIDS cohort in Thailand (2000-2005). AB - This study investigates the survival from the time of diagnosis to death of 98,876 men and 57,316 women between the ages of 15 and 96 years who received medical care at public and private hospitals in Thailand after being diagnosed with symptomatic HIV-positive or AIDS between 2000 and 2005 from all regions of Thailand. Using a retrospective cohort study, risk ratios (RR) and 95% confidence intervals (CI) are estimated by the Cox proportional hazards model adjusting for age, gender, marital status, occupation, region of residence, and year of diagnosis. Significant increased risk of mortality is observed for patients diagnosed with AIDS as compared to those with symptomatic HIV-positive, the risk being approximately twofold for men (RR=2.37, 95% CI=2.27, 2.47), almost threefold for women (RR=2.85, 95% CI=2.69, 3.02), and more than threefold for women in the 15-24-year age group (RR=3.36, 95% CI=2.83, 3.98). The risk also varied from being twofold in the northern region of Thailand (RR=2.23, 95% CI=2.11, 2.35) to being threefold in the northeast (RR=3.05, 95% CI=2.79, 3.32). Our findings of increased risk of mortality for subgroups of the population suggest the need for sustained attention to HIV prevention efforts with support from society at large, and to the early diagnosis and treatment of these patients, especially women and youth. PMID- 20024737 TI - HIV-positive patients' discussion of oral health with their HIV primary care providers in Miami, Florida. AB - Over 90% of HIV-positive persons will have at least one oral manifestation of HIV disease during the course of infection. Clinical guidelines suggest that examination of the oral cavity should be included in initial and interim physical examinations of all HIV-infected patients by their HIV care providers. Clinically significant manifestations of oral disease may impact prescribed treatment regimens. The objective of this analysis was to describe HIV-positive patients' discussion of oral health and dental health with their HIV primary care providers and the correlates of this discussion. We used cross-sectional data from the baseline of a randomized trial testing the efficacy of a risk reduction intervention. Participants were HIV-positive male and female patients attending five HIV primary care clinics in Miami-Dade County, Florida, USA. Overall, 37% of patients did not discuss oral health with their provider. After controlling for age, gender, education, and clinic, odds of discussion of oral health for respondents with five or more primary care visits in the past year were half the odds of those with fewer visits (odds ratio (OR)=0.525, 95% confidence interval (CI): 0.336, 0.821). Odds of discussion for men reporting illicit drug use were 35% of that for non-drug using men (OR=0.353, 95% CI: 0.186, 0.671). Odds of discussion were 1.4 times greater for each additional health topic discussed (e.g., nutrition and smoking) (95% CI: 1.317, 1.544). Given that more than one third of patients reported no discussion of oral health with HIV primary care providers in the past year, there is a need to increase the focus on oral health in the HIV primary care setting. PMID- 20024738 TI - Condom use promotion among isiXhosa speaking women living with HIV in the Western Cape Province, South Africa: a pilot study. AB - The prevalence of HIV infection continues to increase among women in South Africa while there are few interventions specifically targeting condom use promotion in this population. We report the results of an experimental pilot study of a health education intervention aimed at enhancing coping skills and consistent condom use among HIV-positive women attending primary health clinics in the Western Cape province of South Africa. One hundred and twenty women were randomised into the intervention condition or a control condition. Both groups completed an interviewer administered questionnaire that included measures of self-esteem, attitude towards condom use, and self-efficacy towards condom use and negotiating condom use, and provided vaginal swab specimen at baseline and three months after the intervention. Tests for intervention effects at three months while controlling for baseline revealed that only self-esteem was significantly higher in the intervention group relative to the control group. No significant differences were found on measures of coping skills and condom use behaviour. Importantly, incidence for Chlamydia Trachomatis, Neisseria Gonorrhea and Trichomona vaginalis during the study period were significantly lower in the intervention group than the control group. These results are strong indications that this intervention could serve as a basis for the development of potentially effective interventions to reduce STI-related sexual risk behaviours among HIV positive black women in South Africa. PMID- 20024739 TI - Longitudinal association of alcohol use with HIV disease progression and psychological health of women with HIV. AB - We evaluated the association of alcohol consumption and depression, and their effects on HIV disease progression among women with HIV. The study included 871 women with HIV who were recruited from 1993-1995 in four US cities. The participants had physical examination, medical record extraction, and venipuncture, CD4+T-cell counts determination, measurement of depression symptoms (using the self-report Center for Epidemiological Studies-Depression Scale), and alcohol use assessment at enrollment, and semiannually until March 2000. Multilevel random coefficient ordinal models as well as multilevel models with joint responses were used in the analysis. There was no significant association between level of alcohol use and CD4+ T-cell counts. When participants were stratified by antiretroviral therapy (ART) use, the association between alcohol and CD4+ T-cell did not reach statistical significance. The association between alcohol consumption and depression was significant (p<0.001). Depression had a significant negative effect on CD4+ T-cell counts over time regardless of ART use. Our findings suggest that alcohol consumption has a direct association with depression. Moreover, depression is associated with HIV disease progression. Our findings have implications for the provision of alcohol use interventions and psychological resources to improve the health of women with HIV. PMID- 20024740 TI - Which method of adherence measurement is most suitable for daily use to predict virological failure among immigrant and non-immigrant HIV-1 infected patients? AB - In industrialized countries, virological failure occurs more often among HIV infected immigrant patients. Non-adherence is the most credible explanation. We compared adherence of immigrant patients with that of non-immigrant patients in the Netherlands, and investigated which method of adherence measurement is most suitable for daily use to predict virological treatment failure: testing knowledge of the current regimen, a quantitative adherence interview, pharmacy prescription refill ratio (dispensed medication divided by prescribed medication, DM/PM), and plasma drug levels. Included were 61 immigrants and 81 non immigrants. Virological failure did occur more often in immigrants than in non immigrants (19.7% (12/61) versus 8.6% (7/81), p=0.056), especially among previously naive patients (19.6% (11/56) versus 0% (0/54), p<0.01). There were no differences between both groups on any of the four adherence measures. Virological failure was associated with reporting stopping medication when not feeling well (OR=12, 95%CI=1.9-77.7, p=0.02), and, among naive patients, also with a DM/PM < 0.85 (Odds Ratio=5.1, 95%Confidence Interval=1.2-22.3, p=0.03). Although our study confirmed a much higher virological failure rate among immigrants, we were unable to identify clear differences in adherence between immigrants and non-immigrant patient, although virological failure was associated with stopping medication when not feeling well and a low DM/PM. Unstructured treatment interruptions are a likely explanation of the findings. Interventions should be aimed at preventing patients to stop medication. A DM/PM below 0.85 can be indicative for patients who did stop medication and are at risk for virological failure. PMID- 20024741 TI - Knowledge of HIV serostatus and risk behaviour among injecting drug users in Estonia. AB - We used the findings from two, cross-sectional studies of HIV serostatus and risk behaviours to assess the effects of knowledge of HIV serostatus and risk behaviours (relating to sex and injection drug use) among injecting drug users (IDUs). Respondent-driven sampling was used simultaneously at two sites in Estonia (the capital Tallinn, and the second-largest city of Ida-Virumaa County, Kohtla-Jarve). The research tool was an interviewer-administered survey. Biological samples were collected for HIV testing. Participants were categorised into three groups based on HIV testing results and self-report on HIV serostatus: HIV-negative (n=133); HIV-positive unaware of their serostatus (n=75); and HIV positive aware of their serostatus (n=168). In total, 65% of the participants tested positive for HIV. Of those 69% were aware of their positive serostatus. HIV-positive IDUs aware of their serostatus exhibited more risk behaviours than their HIV-positive counterparts unaware of their serostatus or HIV-negative IDUs. Effective prevention of HIV among IDUs should therefore, include programmes to reduce high-risk sexual and drug use behaviours at the public health scale and enhanced prevention efforts focusing on HIV-infected individuals. PMID- 20024742 TI - Discussion and revision of the mathematical modeling tool described in the previously published article "Modeling HIV Transmission risk among Mozambicans prior to their initiating highly active antiretroviral therapy". AB - Mathematical models are increasingly used in social and behavioral studies of HIV transmission; however, model structures must be chosen carefully to best answer the question at hand and conclusions must be interpreted cautiously. In Pearson et al. (2007), we presented a simple analytically tractable deterministic model to estimate the number of secondary HIV infections stemming from a population of HIV-positive Mozambicans and to evaluate how the estimate would change under different treatment and behavioral scenarios. In a subsequent application of the model with a different data set, we observed that the model produced an unduly conservative estimate of the number of new HIV-1 infections. In this brief report, our first aim is to describe a revision of the model to correct for this underestimation. Specifically, we recommend adjusting the population-level sexually transmitted infection (STI) parameters to be applicable to the individual-level model specification by accounting for the proportion of individuals uninfected with an STI. In applying the revised model to the original data, we noted an estimated 40 infections/1000 HIV-positive persons per year (versus the original 23 infections/1000 HIV-positive persons per year). In addition, the revised model estimated that highly active antiretroviral therapy (HAART) along with syphilis and herpes simplex virus type 2 (HSV-2) treatments combined could reduce HIV-1 transmission by 72% (versus 86% according to the original model). The second aim of this report is to discuss the advantages and disadvantages of mathematical models in the field and the implications of model interpretation. We caution that simple models should be used for heuristic purposes only. Since these models do not account for heterogeneity in the population and significantly simplify HIV transmission dynamics, they should be used to describe general characteristics of the epidemic and demonstrate the importance or sensitivity of parameters in the model. PMID- 20024743 TI - HIV prevalence among men and women admitted to a South African public psychiatric hospital. AB - In settings with low seroprevalence, people with severe mental illness have a higher prevalence of HIV infection compared to the general population. In the high-prevalence countries of southern Africa, where the pandemic taxes resources for HIV prevention, care, and treatment, the needs of people with mental illness can be easily overlooked if they are not identified as vulnerable to infection. Yet, few African studies have investigated HIV seroprevalence in psychiatric settings. We systematically examined the HIV seroprevalence among psychiatric patients admitted to a public psychiatric institution in KwaZulu Natal province, South Africa, between 27 July and 14 November 2003. We conducted anonymous testing among 151 patients who were psychiatrically stable and able to give informed consent. Forty patients (26.5%) were HIV-positive; women were more likely to be infected than men (OR 2.74; 95% CI=1.25-6.04; P=0.012). Our findings demonstrate that in the midst of a generalized AIDS epidemic, people with mental illness are also vulnerable and must be included in prevention and treatment efforts. These results underscore the importance of integrated mental health and HIV care in institutional and outpatient mental health settings and affirm the need for detailed HIV risk assessment as a routine part of psychiatric care. Correspondingly, HIV care and treatment programs should be made available to people with psychiatric symptoms. PMID- 20024744 TI - Motivational interviewing by peer outreach workers: a pilot randomized clinical trial to retain adolescents and young adults in HIV care. AB - Youth living with HIV (YLH) are at particularly high risk for poor retention in HIV primary care. This study utilized Motivational Interviewing (MI) to improve youth retention in primary care and compared the fidelity and outcomes of peer outreach workers (POW) to masters level staff (MLS). Eighty-seven YLH were randomized to receive two MI sessions from POW or MLS. YLH were aged 16-29 and 92% were African American. Thirty-seven audiotaped sessions were coded with the Motivational Interviewing Treatment Integrity (MITI) coding system. Retention in care was assessed by review of medical records. POW had higher fidelity on two MITI scales, and did not differ from MLS on remaining three scales. While both groups improved the regularity of primary care appointments, the effect size for POW on retention in care and intervention dose was larger than that of MLS. The results suggest that POW can provide MI with quality comparable to MLS with adequate training and supervision. MI provided by POW to improve retention in health care services may increase the cost-effectiveness of evidence-based practices in urban settings. PMID- 20024745 TI - Trends in primary and secondary abstinence among Kenyan youth. AB - We used data from Kenya Demographic and Health Surveys in 1993, 1998, and 2003 to examine 10-year trends in primary and secondary abstinence among never-married youth aged 15-24 and to explore the role of HIV prevention knowledge, schooling, and contextual factors in affecting their abstinence behaviors. Our analysis shows that both primary and secondary abstinence levels have risen in the past 10 years, with the abstinence levels higher among females than among males. Logistic regression models indicate that knowledge that abstinence can prevent HIV infection was positively associated with the likelihood of practicing abstinence. However, knowledge that condom use can prevent HIV infection was associated with lower abstinence practice. In-school youth were more likely to abstain from sex than those working. Effects of the contextual variables were only significant on the likelihood of primary abstinence among female youth. These findings suggest abstinence programs needed to be gender sensitive and culturally appropriate. PMID- 20024746 TI - Access to highly active antiretroviral therapy (HAART) for women and children in the WHO European Region 2002-2006. AB - Objective. To assess the level of access to highly active antiretroviral therapy (HAART) for women and children in the WHO European Region. Methods. Analysis of data from three national surveys of 53 WHO European Member States. The comparative level of access to HAART for women and children was assessed by comparing the percentage of reported HIV cases with the percentage of HAART recipients in women at the end of 2002 and 2006 and in children at the end of 2004 and 2006. Findings. Overall, the data suggest that there is equivalence of access to antiretroviral therapy by gender and age in Europe. However, in central and eastern Europe women were disproportionately more likely to receive HAART when compared with men in 2006, representing 29% of HIV cases when compared with 39% of HAART recipients in central Europe, and 34% of HIV cases when compared with 42% of HAART recipients in eastern Europe. In comparison with adults, children (<15 years of age) were over-represented among HAART recipients when compared with HIV cases in eastern Europe, accounting for 1% of HIV cases and 9% of people on HAART in 2004 and 1% of HIV cases and 8% HAART recipients in 2006. Conclusion. Access to HAART remains inequitable in terms of gender in central and eastern Europe, favouring women over men, and in terms of age in eastern Europe, favouring children over adults. Despite high and increasing coverage with HAART in many European countries, countries must address how to further increase the number of people on treatment while ensuring equitable access for all population groups in need. PMID- 20024747 TI - The role of HIV testing, counselling, and treatment in coping with HIV/AIDS in Uganda: a qualitative analysis. AB - HIV/AIDS has had a devastating impact at individual, household and community levels. This qualitative research investigates the role of HIV voluntary counselling and testing (VCT) and treatment in enabling HIV-positive Ugandans to cope with this disease. Twelve predetermined focus group discussions (FGDs) were conducted; six with men and six with women. Half of the men and women's groups were receiving antiretroviral therapy (ART) and half were not. An FGD was held with the health care providers administering ART. Testing for HIV was perceived as soliciting a death warrant. Participants affirmed that the incentive for testing was the possibility of accessing free ART. They described experiencing gender-variant stigma and depression on confirming their HIV status and commended the role of counselling in supporting them to adopt positive living. For those receiving ART, counselling reinforced treatment adherence. The findings also revealed gender differences in treatment adherence strategies. ART was described to reduce disease symptoms and restore physical health allowing them to resume their daily activities. Additionally, ART was preferred over traditional herbal treatment because it had clear dosages, expiry dates and was scientifically manufactured. Those that were not receiving ART bore myths and misconceptions about the effectiveness and side effects of ART, delaying the decision to seek treatment. Stigma and the attached concern of HIV/AIDS-related swift death, is a major barrier for VCT. Based on this study's findings, ensuring the provision of quality assured and gender conscious VCT and ART delivery services will enhance positive living and enforce compliance to ART programmes. PMID- 20024748 TI - Comparing prevalence of HIV-related behaviors among female injecting drug users (IDU) whose regular sexual partner was or was not IDU in Sichuan and Yunnan Provinces, China. AB - Injecting drug users made up a high percentage of HIV cases in China; partners of IDU are hence, at very high risk of HIV transmission. The goal of this study is to compare characteristics and prevalence of injecting drug use and sex-related risk behaviors among female IDU who were or were not having a regular sex partner (RSP) with concordant IDU status. Data were obtained from all female IDU respondents (n=1414) surveyed in the behavioral surveillance surveys as conducted during 2003 through 2006 in Sichuan and Yunnan, China. The findings from this research reveal that about half of the female IDU respondents belonged to the concordant group (50.4%). As compared to members of the discordant group, such concordant respondents were significantly more likely to have practiced different types of studied syringe sharing behaviors (adjusted odds ratios AOR=1.34-2.23, p<0.05), to have served as female sex workers in the last month (AOR=1.24, p<0.05), or to have used some particular types of HIV-related prevention services (AOR=1.36-1.60, p<0.05) but were less likely to be always using new needles for drug injection in the last month (AOR=0.54, p<0.05) or to have used a condom in the last episode of sex with their RSP (AOR=0.64, p<0.01). The findings suggest that concordant IDU status is very common among RSP of IDU. The risk for HIV transmission between concordant RSP seems to be very high and the prevention services seem unable to alleviate the risk. Specific programs targeting concordant IDU couples are greatly warranted. PMID- 20024749 TI - Community interventions providing care and support to orphans and vulnerable children: a review of evaluation evidence. AB - Children affected by HIV in their families and communities face multiple risks to their health, education and psychosocial wellbeing. Community interventions for children who have been orphaned or rendered vulnerable take many forms, including educational assistance, home-based care, legal protection and psychosocial support. Despite a recent influx of funding for programme implementation, there exists little evidence to inform policymakers about whether their investments are improving the lives of vulnerable children and meeting key benchmarks including the Millennium Development Goals. This paper reviews the current evidence base on evaluations of community interventions for orphans and vulnerable children (OVC) in high HIV-prevalence African settings, focusing on studies' methodologies. Sources reviewed include published research studies and evidence from the unpublished programmatic "grey literature" located through database and internet searches. A total of 21 studies, varying in scope and generalisability, were identified. Interventions reviewed address children's wellbeing through various strategies within their communities. Evaluation methodologies reflect quantitative and qualitative approaches, including surveys (with and without baseline or comparison data), costing studies, focus groups, interviews, case studies, and participatory review techniques. Varied study methodologies reflect diverse research questions, various intervention types, and the challenges associated with evaluating complex interventions; highlighting the need to broaden the research paradigm in order to build the evidence base by including quasi-experimental and process evaluation approaches, and seeking further insights through participatory qualitative methodologies and costing studies. Although findings overall indicate the value of community interventions in effecting measurable improvements in child and family wellbeing, the quality and rigour of evidence is varied. A strategic research agenda is urgently needed to inform resource allocation and programme management decisions. Immediate imperatives include building local technical capacity to conduct quantitative and qualitative evaluation research, and strengthening monitoring and evaluation systems to collect process and outcome data (including costing) on key support models. Donors and implementers must support the collection of sound empirical evidence to inform the development and scale-up of OVC programmes. PMID- 20024750 TI - No differences in quality of life between men and women undergoing HIV antiretroviral treatment. Impact of demographic, clinical and psychosocial factors. AB - INTRODUCTION: The health-related quality of life (HRQL) is therefore a highly important tool for the evaluation of the health status of HIV positive patients. However, very little information is available about the differences between HRQL in men and women. The aim of this study is to analyse gender differences in HRQL in HIV positive patients. METHODOLOGY: A sample group of 540 patients infected with HIV and undergoing antiretroviral treatment was studied. All of these patients attended outpatient appointments in the infectious disease units of four hospitals in the southern Spanish region of Andalusia. HRQL was measured using the MOS-HIV health survey, which had been validated for use in this setting. RESULTS: One hundred forty-nine women (26.6%) and 411 men (73.4%) took part in the study. On the whole, the women were younger, more likely to be married and had more children. The men, on the other hand, were more likely to present with coinfection with hepatitis C, have a history of intravenous drug use, be undergoing methadone treatment or have lower levels of CD4. Both the overall average scores and the scores for specific dimensions were higher in women than in men. When these scores were adjusted using different clinical variables, these differences were not statistically significant. Amongst the women, psychic morbidity, longer periods of time as HIV carriers and low levels of social support were linked to lower MHS and PHS scores. An independent link was established between older age and a lower PHS score. Amongst the men, psychic morbidity, difficulty in taking tablets, intravenous drug use and low levels of social support were linked to lower PHS and MHS scores. AIDS and a higher number of antiretroviral medicines were linked to a lower PHS score. CONCLUSIONS: By carrying out an analysis according to gender it is possible to see different factors associated with a poorer quality of physical and mental health. This makes it possible to identify areas which could be developed and to optimise results in terms of patients' health. PMID- 20024752 TI - Using the Theory of Planned Behavior to investigate condom use behaviors among female injecting drug users who are also sex workers in China. AB - Female injecting drug users who are sex workers (IDUFSWs) is a strategic "bridge population" for HIV transmission. Goals of the study were to investigate condom use behaviors during commercial sex among IDUFSWs using the Theory of Planned Behavior (TPB), and to investigate moderating effects that modify the strength of associations between the TPB-related variables and inconsistent condom use during commercial sex. A total of 281 non-institutionalized IDUFSWs were recruited using snowball sampling method. Anonymous face-to-face interviews were administered by trained doctors. The results showed that the prevalence of inconsistent condom use during commercial sex in the last six months was 64%. After adjusting for some significant background variables (e.g. main venue of sex work), all associations between the five TPB-related variables and the studied condom use variable were statistically significant (Odds Ratio (OR) = 0.43-0.68, p<0.001). In the hierarchical nested models, three background variables (age, venue of sex work, and ever used HIV-related interventions) entered in the first step (-2LL = 294.98, p<0.001) and the Social Norm Scale, the Perceived Behavioral Control Scale and the Behavioral Intention Scale were selected by the second step (OR = 0.67 - 0.72, p<0.01; -2LL = 160.99, p<0.001). Significant moderating effects between some TPB-related variables (Positive Condom use Attitude Scale and Behavioral Intention Scale) and duration of sex work and duration of drug use were also reported. The results highlighted the potential of using the TPB to better understand condom use behaviors in IDUFSWs in China. Theory-based research and intervention work should be developed in China in the future. PMID- 20024751 TI - A systematic review comparing antiretroviral adherence descriptive and intervention studies conducted in the USA. AB - We examined the extent to which studies aimed at testing interventions to improve antiretroviral adherence have targeted the facilitators of and barriers known to affect adherence. Of the 88 reports reviewed, 41 were reports of descriptive studies conducted with US HIV-positive women and 47 were reports of intervention studies conducted with US HIV-positive persons. We extracted from the descriptive studies all findings addressing any factor linked to antiretroviral adherence and from the intervention studies, information on the nature of the intervention, the adherence problem targeted, the persons targeted for the intervention, and the intervention outcomes desired. We discerned congruence between the prominence of substance abuse as a factor identified in the descriptive studies as a barrier to adherence and its prominence as the problem most addressed in those reports of intervention studies that specified the problems targeted for intervention. We also discerned congruence between the prominence of family and provider support as factors identified in the descriptive studies as facilitators of adherence and the presence of social support as an intervention component and outcome variable. Less discernible in the reports of intervention studies was specific attention to other factors prominent in the descriptive studies, which may be due to the complex nature of the problem, individualistic and rationalist slant of interventions, or simply the ways interventions were presented. Our review raises issues about niche standardization and intervention tailoring, targeting, and fidelity. PMID- 20024753 TI - Depression and symptomatic response among HIV-infected drug users enrolled in a randomized controlled trial of directly administered antiretroviral therapy. AB - BACKGROUND: Depression is highly prevalent among HIV-infected injection drug users (IDUs) and has been associated with poor adherence to antiretroviral therapy and increased morbidity and mortality. Factors associated with changes in depressive symptoms among this group receiving antiretroviral treatment that have not been extensively evaluated. METHODS: This post-hoc analysis of prospective clinical trial analyzes the factors associated with changes in depressive symptomatology using the Center for Epidemiologic Studies of Depression Scale (CES-D) among HIV-infected IDUs enrolled in a prospective, six months randomized controlled trial of directly administered antiretroviral therapy (DAART) versus self-administered therapy. RESULTS: Of the 127 evaluable IDUs enrolled in the study, 89 subjects (70%) had complete six-month follow-up data. Of these, 58 (63%) met baseline criteria for severe or major depressive disorder (MDD) using the CES-D. CES-D scores improved significantly from baseline to six months overall for the 89 subjects (p=0.01) and for the 58 who had MDD with six-month data (p=0.001). Using multiple regression, an improvement in CES-D score was independently associated with: (1) increase in CD4 count; (2) increase in adherence; (3) non-Caucasian race; and (4) older age. Worsening in CES-D score was associated with: (1) increase in HIV-1 RNA levels; (2) homelessness; (3) poor self-efficacy; (4) active drug use; and (5) male gender. Factors not correlated with changes in CES-D were receipt of DAART, engagement in drug treatment, use of antidepressant medication, and employment. Using generalized estimating equation modeling, factors that remained positively associated with improvements in CES-D score were absence of drug use at six months, having housing, higher self efficacy, increase in CD4 count and increases in adherence. CONCLUSION: Improvements in depressive symptoms could occur with improvement of alterable factors that are associated with strengthening adherence such as linkages to case management, mental health and substance abuse treatment services as well as through enhancement of social stabilization factors through social support and supportive housing. PMID- 20024754 TI - The relative risk of HIV among IDUs not in treatment in Malaysia. AB - Despite the growing HIV threat among injecting drug users (IDUs) in Malaysia, there is a dearth of information on their HIV risk behaviour. This study focused on identifying specific risk behaviours that distinguished HIV positive IDUs from those who were not. For the first time, data on IDUs not in treatment were obtained through a cross-sectional survey of 526 subjects recruited from five selected cities across peninsular Malaysia. A structured questionnaire and face to-face interviews were utilised to collect detailed information on their drug use practices and sexual behaviours. On-site serological testing determined their HIV and hepatitis C status. The findings indicated that ethnic Malays, who are also Muslims, form the majority of IDUs not in treatment. Bivariate analysis identified six risk factors associated with HIV seropositivity: being 44 years or younger; not holding a regular job; initiating drug use at age 23 or younger; being a morphine user; sharing injecting equipment and having multiple-sex partners. However, only the last two remained significant in multivariate analysis. That sharing contaminated injecting equipment is a significant risk factor strongly justifies the widening of the pilot needle and syringe exchange programme initiated hesitantly in late 2005 as a reaction to the worsening HIV/AIDS situation. Condom use, though not independently significant, remains important because consistent and wider use could neutralise the second risk factor--having multiple-sex partners. The finding that injecting drug use is increasingly occurring in groups underscores the need for outreach programmes that emphasise safe injecting practices in group settings. In addition, counsellors should endeavour to convince drug users to enter treatment since being in treatment appears to reduce risk behaviours. Finally, conservative Muslim unease about harm reduction must be assuaged quickly since Malay Muslims form the majority of IDUs not in treatment. PMID- 20024755 TI - Feasibility of using audio computer-assisted self-interview (ACASI) screening in routine HIV care. AB - We evaluated the feasibility of implementing audio computer-assisted self interviews (ACASI) as part of routine clinical care at two community hospital based HIV clinics in New York City. Between June 2003 and August 2006, 215 patients completed 1001 ACASI sessions in English or Spanish prior to their scheduled clinical appointments. Topics covered included antiretroviral therapy adherence, depression symptoms, alcohol and drug use, and condom use. Patients and providers received feedback reports immediately after each session. Feasibility was evaluated by quantitative analysis of ACASI responses, medical chart reviews, a brief patient questionnaire administered at the conclusion of each computer session, patient focus groups, and semi-structured provider interviews. ACASI interviews frequently identified inadequate medication adherence and depression symptoms: at baseline, 31% of patients reported < or =95% adherence over the past three days and 52% had symptoms of depression (CES-D score > or =16). Substance abuse problems were identified less frequently. Patients were comfortable with the ACASI and appreciated it as an additional communication route with their providers; however, expectations about the level of communication achieved were sometimes higher than actual practice. Providers felt the summary feedback information was useful when received in a timely fashion and when they were familiar with the clinical indicators reported. Repeated ACASI sessions did not have a favorable impact on adherence, depression, or substance use outcomes. No improvements in HIV RNA suppression were observed in comparison to patients who did not participate in the study. We conclude that it is feasible to integrate an ACASI screening tool into routine HIV clinical care to identify patients with inadequate medication adherence and depression symptoms. Repeated screening was not associated with improved clinical outcomes. ACASI screening should be considered in HIV clinical care settings to assist providers in identifying patients with the greatest need for targeted psychosocial services including adherence support and depression care. PMID- 20024756 TI - Can you hear me now? Limited use of technology among an urban HIV-infected cohort. AB - Recent studies support technology-based behavioral interventions for individuals with HIV. This study focused on the use of cell phone and internet technologies among a cohort of 515 HIV-infected individuals. Socio-demographic and clinic data were collected among individuals presenting at an urban Midwestern university HIV clinic in 2007. Regular internet usage occurred more often with males, Caucasians, those who were employed, had higher income, and were more educated. Higher levels of education and income >$10,000 predicted regular usage when controlling for race, employment, and gender. Cell phone ownership was associated with being Caucasian, employed, more educated, and salary >$10,000. Employment was the only predictor of owning a cell phone when controlling for income, race, and education. Individuals who were <40 years of age, employed, and more educated were more likely to know how to text message. Employment and post-high school education predicted knowledge of text messaging, when controlling for age. Disparities among internet, cell phone, and text messaging usage exist among HIV infected individuals. PMID- 20024757 TI - Stigma, social support, and depression among people living with HIV in Thailand. AB - BACKGROUND: People living with HIV/AIDS (PLWHA) in Thailand face tremendous challenges, including HIV-related stigma, lack of social support, and mental health issues such as depression. This study aims to examine complex relationships among demographics, HIV-related stigma, and social support and their impact on depression among PLWHA in Thailand. METHODS: This study uses data collected in northern and northeastern Thailand. A total of 408 PLWHA were recruited and interviewed in 2007. HIV-related stigma was measured by two subscales: "Internalized Shame" and "Perceived Stigma." Based on correlation analyses, hierarchical multiple regression models were used to examine the predictors of depression, social support, and HIV-related stigma, controlling for demographic characteristics. RESULTS: Correlational analysis revealed that depression was significantly associated with both dimensions of stigma: internalized shame and perceived stigma. Self-reported emotional social support was negatively associated with depression. We found that internalized shame and emotional social support were significant predictors of depression after controlling for gender, age, income, and education. CONCLUSIONS: HIV-related stigma has a negative impact on psychological wellbeing of PLWHA in Thailand, and emotional social support remains a protective factor against depression. Intervention developers and clinicians working with PLWHA may find it useful to incorporate the association between stigma and depression into their programs and treatments, and to address social support as a protective effect for the mental health of PLWHA. PMID- 20024758 TI - Gender differences in unsafe sexual behavior among young people in urban Mali. AB - We developed gender-specific explanatory models for unsafe sexual behavior among unmarried young people living in urban West Africa using a culturally adapted instrument which addresses personal, relational and socio-cognitive factors. Data were collected on condom use and number of sexual partners, and on their potential determinants, using in-person interviews on a heterogeneous sample of in-school and out-of-school young women (n=185) and (n=214) men who reported ever having had sexual intercourse. Recruitment was done at various sites in 21 randomly selected neighborhoods in Bamako. Bivariate analysis and multivariate logistic regressions were conducted to identify determinants of lack of condom at last sex and having more than one partner in the last six months and to test for interactions with gender. Similar percentages of men (40%) and women (46%) reported not using a condom at last sex. However, more men (64%) reported multiple partnering in the last six months than women (32%). Our findings suggest that the context of sexual debut, social status, relation to peers and family as well as attitudinal, normative and behavioral control constructs may influence young men and women's sexual behavior, but that pathways leading to unsafe sex are different across genders. Our findings also show that factors associated with condom use and sexual partnering are distinct. Earlier sexual debut seems to be risk-inducing in women while risk-reducing in men. Poor communication with peers and receiving money from sexual partner were associated with lack of condom use in women. High behavioral control was associated with fewer sexual partners in men. Determinants of unsafe sexual behavior were found at the social, interpersonal and individual levels for both men and women, but notable gender differentials existed. These findings underscore the importance of addressing gender as a crucial factor shaping HIV-risk profiles. PMID- 20024759 TI - Factors influencing adherence to anti-retroviral treatment in children with human immunodeficiency virus in South India--a qualitative study. AB - The aim of the study was to gain a deeper understanding of the factors that influence adherence to Anti-Retroviral Treatment (ART) in a paediatric population in South India. Semi-structured interviews, guided by a questionnaire based on literature review, were undertaken with 14 primary caregivers of children with Human Immunodeficiency Virus (HIV) on ART focussing on the factors influencing adherence and non-adherence. Adherence to ART was assessed by caregiver reported adherence for the last three days. The framework approach was used for analysis of the transcribed interviews. Adherence behaviour in children taking ART was influenced by medication, child, caregiver, family and healthcare service related factors. Medication related factors such as palatability, formulation, and difficult regime were among the common reasons for non-adherence. While a supportive family appeared to contribute to adherence, some caregivers were reluctant to inform their extended family members for fear of isolation. All participants were strongly appreciative of the study centre's model of care and this had a positive effect on adherence behaviour. From the public health point of view, the respondents' reluctance to access publicly funded HIV care services is a cause for concern. Almost all caregivers were sceptical of the quality of free ART medications provided by public services, indicating an urgent need for appropriate education to alleviate their concerns. The cost of ART was reported by caregivers as an important barrier, particularly by those with low income. Most caregivers recommended wider access to free ART, availability of child friendly medications and adherence counselling as important interventions to improve adherence. All caregivers in this study reported some difficulty in sustaining adherence in their children. Further research on improving adherence in children by tailoring interventions to the specific needs of caregivers and their families is warranted. PMID- 20024760 TI - Household impacts of AIDS: using a life course approach to identify effective, poverty-reducing interventions for prevention, treatment and care. AB - A life course approach was used to assess household level impacts and inform interventions around HIV risk and AIDS vulnerability across seven major age related stages of life. Our focus was sub-Saharan Africa. We provided a qualitative review of evidence from published literature, particularly multicountry reviews on impacts of AIDS, on determinants of risk and vulnerability, and reports of large surveys. Areas of potential stress from birth to old age in households affected by AIDS, and interventions for dealing with these specific stresses were identified. While specific interventions for HIV are important at different stages, achieving survival and development outcomes demands a wider set of health, social security and development interventions. One way to determine the priorities amongst these actions is to give weighting to interventions that address factors that have latent impacts later in life, which interrupt accumulating risk, or that change pathways to reduce the risk of both immediate and later stress. This qualitative review suggested that interventions, important for life cycle transitions in generalised epidemics where HIV risk and AIDS vulnerability is high, lie within and outside the health sector, and suggested examples of such interventions. PMID- 20024761 TI - Condom use: exploring verbal and non-verbal communication strategies among Latino and African American men and women. AB - A growing body of literature provides evidence of a link between communication with sexual partners and safer sexual practices, including condom use. More research is needed that explores the dynamics of condom communication including gender differences in initiation, and types of communication strategies. The overall objective of this study was to explore condom use and the dynamics surrounding condom communication in two distinct community-based samples of African American and Latino heterosexual couples at increased risk for HIV. Based on 122 in-depth interviews, 80% of women and 74% of men reported ever using a condom with their primary partner. Of those who reported ever using a condom with their current partner, the majority indicated that condom use was initiated jointly by men and women. In addition, about one-third of the participants reported that the female partner took the lead and let her male partner know she wanted to use a condom. A sixth of the sample reported that men initiated use. Although over half of the respondents used bilateral verbal strategies (reminding, asking and persuading) to initiate condom use, one-fourth used unilateral verbal strategies (commanding and threatening to withhold sex). A smaller number reported using non-verbal strategies involving condoms themselves (e.g. putting a condom on or getting condoms). The results suggest that interventions designed to improve condom use may need to include both members of a sexual dyad and focus on improving verbal and non-verbal communication skills of individuals and couples. PMID- 20024762 TI - HIV-related behaviors among black students attending Historically Black Colleges and Universities (HBCUs) versus white students attending a traditionally white institution (TWI). AB - Recent data indicated the unexpected movement of the HIV epidemic in the Southeastern US and the transmission among college students especially in black campuses. The current study is the first one attempted to assess and compare HIV related behaviors among black students attending Historically Black Colleges and Universities (HBCUs) and white students attending a traditionally white institution (TWI). Black students were recruited from HBCUs (n=222) and white students from a TWI (n=335) via online surveys. The majority of students in both samples were between 18 and 24 years old (mean = 20.65) and self-identified as heterosexual oriented (91%), although the HBCU sample revealed slightly higher proportion of females (81.1% vs. 72.5%). Data indicated different behavioral patterns. Although, both groups reported similar prevalence of oral and anal sex; after adjusting for age and gender, HBCU blacks were more likely to have had vaginal sex (OR = 1.7) and at younger age (16.3 vs. 17.5 years), more likely to have had Sexually Transmitted Infections (OR = 4.4), have been or gotten someone pregnant (OR = 3.6). They were, however, less likely to use alcohol before sex of any type, and more likely to have been tested for HIV (OR = 8.4), ask partner's status (OR = 3.8) or being asked of status (OR = 2.9). Comparing with TWI whites, HBCU blacks also perceived higher risk of HIV infection, higher peer norms toward vaginal and anal sex. The findings showed HBCU students taking more protective behaviors in some aspects while practicing riskier behaviors in others. Results have implications on developing tailored HIV behavioral interventions. PMID- 20024763 TI - Characteristics of sexual partnerships, not just of individuals, are associated with condom use and recent HIV infection in rural South Africa. AB - Characteristics of sexual partnerships, as well as those of the individuals involved, might influence the use of condoms and risk of HIV transmission. We set out to identify characteristics of non-spousal sexual partnerships associated with condom use at last sex in the previous year and HIV infection in the previous three years among sexually active young people in rural South Africa. We conducted an analysis of follow-up data (collected in 2004) from a cohort of 14 35-year old men and women recruited to a cluster-randomised trial. Data on 1647 non-spousal sexual partnerships during the previous year were reported in 2004 and analysed alongside new HIV infections over the previous three years among 762 individuals who were HIV-negative in 2001. Structured interviews elicited information on sexual behaviour. HIV serostatus was assessed through oral-fluid ELISA. Condom use at last sex was reported for 615/1647 non-spousal sexual partnerships (37.3%) and was more commonly reported by individuals who were younger, more educated and aware of their HIV status. Condom use was more common in casual partnerships, those where the male partner was younger, where sex was less frequent and where the respondent believed the partner to have other sexual contacts. New HIV infection in the last three years was identified for 87/762 individuals (11.4%) and was more common among females and those out of school. Infection risk was associated with the age of the partners and was less common among individuals reporting less frequent intercourse in the previous year. Characteristics of sexual partnerships, as well as those of individuals, are important determinants of condom use and risk of HIV infection. Male characteristics may be particularly important because of their greater capacity to make decisions about HIV prevention. Established non-spousal sexual partnerships are an increasingly important context for HIV transmission in this setting. PMID- 20024764 TI - Managing identity impacts associated with disclosure of HIV status: a qualitative investigation. AB - Disclosure of HIV status to potential and current sex partners by HIV-positive people (HIVPP) is a complex issue that has received a significant amount of attention. Research has found that disclosure depends upon the evaluation by HIVPP of potential benefits and risks, especially of the risks stemming from the profound social stigma of HIV and AIDS. Drawing on concepts from Goffman's classic stigma theory and Anderson's more recently developed cultural-identity theory of drug abuse, we analyzed data from in-depth, post-intervention qualitative interviews with 116 heterosexually active, HIV-positive injection drug users enrolled in a randomized trial of a behavioral intervention to prevent HIV transmission. We explored how disclosure experiences lead to "identity impacts" defined as: (1) identity challenges (i.e. interactions that challenge an individual's self-concept as a "normal" or non-deviant individual); and (2) identity transformations (i.e. processes whereby an individual comes to embrace a new identity and reject behaviors and values of an old one, resulting in the conscious adoption of a social and/or public identity as an HIV-positive individual). Participants engaged in several strategies to manage the identity impacts associated with disclosure. Implications of these findings for research and prevention programming are discussed. PMID- 20024766 TI - Increased HIV disclosure three months after an online video intervention for men who have sex with men (MSM). AB - A behavioral intervention for men who have sex with men (MSM) was created for online delivery. The nine-minute video drama "The Morning After" (www.hivbigdeal.org) was designed to promote critical thinking about HIV risk. MSM were recruited for the evaluation through banner ads on a subscription-based gay sexual meeting web site. Participants viewed the intervention online and completed online behavioral questionnaires at enrollment and three month follow up. Each participant served as his own control. No participant incentives were provided. Follow-up was completed by 522 (54%) of 971 eligible men. Men completing follow-up were similar to those who did not by age and prevalence of unprotected anal intercourse (UAI) but reported more lifetime sex partners. They also differed somewhat by race, education, and HIV testing. In the three months after the intervention, men were significantly more likely to disclose HIV status to partners (odds ratio [OR] = 3.37, p<0.001) and less likely to report a casual partner or UAI in their most recent sexual encounter than at baseline. Of 120 men HIV tested during follow-up, 17 (14%) reported being HIV seropositive. These findings show that MSM at high risk for HIV will participate in a brief online video intervention designed to engage critical thinking and that significant, self-reported increases in HIV disclosure and decreases in risk behavior occurred three months after MSM viewed the video. PMID- 20024768 TI - Does age affect sexual behaviour among gay men in Sydney, Melbourne and Brisbane, Australia? AB - Concern internationally that young gay men are at increased risk of HIV infection has not been reflected in earlier findings in Australia where younger men have not been found to be at increased likelihood to engage in unprotected anal intercourse with casual partners (UAIC). There has, however, been little attention paid to the issue of age in relation to HIV risk behaviour in Australia in recent years. In 2007, among men who completed Gay Community Periodic Survey questionnaires in Sydney, Melbourne and Brisbane, Australia, younger men were more likely to report being in relatively short-term monogamous relationships than were their older counterparts. They were also less likely to know their own or their partners' HIV serostatus. Men aged less than 25 years reported fewer recent partners and were less likely to report sex with casual male partners in the previous six months (p<0.001). Younger men were also less likely to engage in group sex. Approximately, one-quarter of the sample reported engaging in any UAIC in the previous six months during each year of the survey. Younger men were no less likely to report UAIC overall, but they were somewhat more likely to report taking the receptive position during UAIC. While age is a consideration in the assessment of risk of HIV transmission among gay men, this risk is dependent upon the context in which it occurs: Age mixing may be an important consideration in understanding HIV risk among young gay men. PMID- 20024767 TI - Development of a treatment optimism scale for HIV-positive gay and bisexual men. AB - Potential items to be included in an HIV Treatment Optimism scale were reviewed by 17 HIV-positive gay and bisexual men (GBM), resulting in a 21-item test instrument. After pilot testing, data were collected from a multi-city sample of high-risk HIV-positive GBM (n=346), who were currently on treatment and were recruited to attend a two-day sexual health seminar. The scale items were analyzed utilizing Principal Components Analysis and reliability testing. The factor analysis resulted in the development of three separate scales. The Susceptibility scale contained 10 items associated with a belief that HIV is less transmissible while on HIV treatment. The Condom Motivation scale contained five items addressing a decreased motivation to use condoms while on treatment and the Severity scale contained four items associated with a decreased sense of the severity of an HIV diagnosis. Reliability coefficients (alpha) and mean inter item correlations (M) for the three scales were acceptable (Susceptibility, alpha = 0.86, M=0.39; Condom Motivation, alpha = 0.84, M = 0.50; Severity, alpha = 0.71, M=0.37). Combined as one scale,the reliability coefficient was respectable (alpha = 0.76), but the mean inter-item correlation was 0.14. Based on this analysis, use of a single measure was not supported and three separate scales were developed. The scales were equivalent across racial groups except White men were more like to report a decreased motivation to use condoms compared to Black or Latino men. Three separate scales addressing beliefs about the transmissibility of HIV while on treatment (Susceptibility), the quality of life while on HIV treatment (Severity) and the motivation to use condoms consistently while on treatment (Condom Motivation) may be better markers for assessing optimistic beliefs about HIV treatment among HIV-positive GBM. PMID- 20024769 TI - HIV and tuberculosis in Durban, South Africa: adherence to two medication regimens. AB - Given that antiretroviral (ARV) medication adherence has been shown to be high in resource-limited countries, the question remains as to whether adherence will remain at that level as medications become more widely available. Comparing adherence to tuberculosis (TB) medications, which have been readily available, and ARV medications may help to indicate the likely future adherence to ARVs as access to these medications becomes more widespread. This study examined sense of coherence, social support, symptom status, quality of life, and adherence to medications in two samples of individuals being treated either for TB or human immunodeficiency virus/acquired immunodeficiency syndrome (HIV/AIDS) at clinics in Durban, South Africa. Findings revealed the distinctive socio-economic backgrounds of the two cohorts. Although there were significant differences with regard to the psychosocial variables, there were no significant differences by the two samples in adherence to medications as well as adherence to appointments. Given the self-selected nature of the participants in this study, namely those able to attend clinic, as well as those likely to be adherent to ARVs, there is every reason for caution in the interpretations of these findings. As access to ARV medications becomes more widely available in South Africa, the question remains as to whether such high adherence will be maintained given the constraints of access to food and other basic necessities. PMID- 20024770 TI - Social determinants of mixed feeding behavior among HIV-infected mothers in Jos, Nigeria. AB - Mixed feeding confers excess risk of mother-to-child transmission (MTCT) of HIV compared with exclusive breastfeeding and exclusive formula feeding. We undertook a qualitative and quantitative cross-sectional survey to identify the social determinants of mixed feeding among a subset of the 469 HIV-infected women enrolled in a MTCT prevention program in Jos, Nigeria. Formula was provided free of-cost. Of the 91 participants, 68 (75%) exclusively formula fed, 7 (8%) exclusively breastfed, and 16 (18%) practiced mixed feeding. Of the mixed feeding women, seven primarily formula fed and nine primarily breastfed. Women who primarily formula fed described family pressure as the reason for mixed feeding, while women who primarily breastfed reported insufficient breast milk. In a multivariate analysis, lack of partner support of the feeding decision predicted mixed feeding behavior (OR: 4.2; 95% CI: 1.2-14.9; p=0.03). Disclosure of HIV status was significantly correlated (p<0.001) with partner support. HIV prevention interventions aimed at reducing mixed feeding should encourage supportive partner relationships that facilitate disclosure of HIV status. Attention should also be made to the differing pressures faced by women attempting to exclusively breast feed and exclusively formula feed. PMID- 20024771 TI - Access to HIV/AIDS care for mothers and children in sub-Saharan Africa: adherence to the postnatal PMTCT program. AB - Despite scale up of perinatal prevention of mother-to-child transmission (PMTCT) of HIV interventions, postnatal continuity of comprehensive HIV/AIDS care, for both the mother and baby, remains a challenge in developing countries. We determined adherence to the postnatal PMTCT program (PN-PMTCT) and the associated factors among mothers at a public urban hospital in Uganda. We interviewed HIV positive postnatal mothers on discharge and we determined adherence to PN-PMTCT by the proportion of mothers that honored their return appointments by the end of eight weeks postpartum. We had focus group discussions to assess factors that influence adherence to PN-PMTCT. Of 289 mothers, only 110 (38%) adhered to PN PMTCT. Previous attendance of a routine postnatal review and having access to a phone were significantly associated with adherence to PMTCT among mothers older than 25 years (odds ratio (OR) 3.6 (95% confidence interval (CI); 1.2-10.4)) and (OR 3.1 (95% CI; 1.3-7.1)), respectively. On the other hand, Christianity (OR 3.2 (95% CI; 1.1-9.0)) was significantly associated with adherence to PN-PMTCT among mothers below 25 years of age. Mothers' perceived benefits of the PN-PMTCT program, easy access to the program, and presence of social support from a spouse were important motivators for mothers to adhere to PN-PMTCT. Even with improved antenatal and intra-partum PMTCT services, only a third of the HIV-infected mothers adhered to the PN-PMTCT program. Mothers who previously attended a routine postnatal care were 3.6 fold more likely to adhere to PN-PMTCT. We recommend strategies to increase mothers' adherence to PN-PMTCT interventions in order to increase access to HIV/AIDS care for mothers and children in sub-Saharan Africa. PMID- 20024772 TI - Measuring the quality of life of residents in SADC communities affected by HIV. AB - About a third of all people living with HIV are found in Southern Africa. Local and regional organisations work in and with communities subjected to high HIV prevalence rates. It is necessary to measure the Quality of Life (QoL) of community members affected by HIV to improve monitoring and evaluation of project impact. This study aimed to measure the QoL and to investigate the QoL scale as a rapid cross-cultural assessment tool. QoL surveys were conducted in 14 community sites in seven countries, including: Lesotho, Malawi, Mozambique, South Africa, Swaziland, Zambia and Zimbabwe. These communities are amongst the most severely affected by the HIV/AIDS epidemic in the region. Face-to-face interviews, using a structured questionnaire, were conducted by teams of trained fieldworkers. The studies included 2176 participants, including 37% males. Results indicated evidence of the heavy burden of the disease. More than 30% of the households had a member who was ill for more than three months and more than 10% of the households lost a member after an illness of three months in the past 12 months. More than 35% of the households housed an orphan. Unemployment rates in the communities were very high. QoL was measured using four domains: General Health, Psychological Wellbeing, Social Relationships and Environmental factors. General Health and Environmental quality consistently rated lower than Psychological Wellbeing and Social Relationships. The QoL questionnaire seems to be appropriate in measuring QoL of community members cross-culturally. Results will enable programmes and organisations to tailor and develop projects. It will assist organisations to continuously monitor activities and its impact. PMID- 20024773 TI - Adherence to antiretroviral therapy: a survey of factors associated with medication usage. AB - The use of highly active antiretroviral therapy (HAART) has decreased morbidity and mortality for people living with HIV/AIDS, but adherence to HAART is a critical factor in successful treatment. Adherence to medication is a complex and poorly understood behavior. A survey was undertaken to evaluate subjective correlates of adherence and non-adherence based on previously distilled themes from a qualitative study of adherence. A 60-question survey was completed by patients in the outpatient clinic setting regarding demographics, CD4 cell count, viral load, adherence, and screening questions about medication usage and attitudes toward HIV. Ninety-eight adherent and 34 non-adherent patients completed the survey. After logistic regression analysis, several questions appeared to be the main predictors of non-adherence: Have you ever thought of having HIV as a "punishment?" Do you feel that your medicines are hard to take? Do you believe the medicines for HIV that you take are working for you? These questions may be helpful in the development of a clinically useful screening tool to assess patients at risk for non-adherence. PMID- 20024774 TI - Social support and delays seeking care after HIV diagnosis, North Carolina, 2000 2006. AB - Many adults in the USA enter primary care late in the course of HIV infection, countering the clinical benefits of timely HIV services and missing opportunities for risk reduction. Our objective was to determine if perceived social support was associated with delay entering care after an HIV diagnosis. Two hundred and sixteen patients receiving primary care at a large, university-based HIV outpatient clinic in North Carolina were included in the study. Dimensions of functional social support (emotional/informational, tangible, affectionate, and positive social interaction) were quantified with a modified Medical Outcomes Study Social Support Scale and included in proportional hazards models to determine their effect on delays seeking care. The median delay between diagnosis and entry to primary care was 5.9 months. Levels of social support were high but only positive social interaction was moderately associated with delayed presentation in adjusted models. The effect of low perceived positive social interaction on the time to initiation of primary care differed by history of alcoholism (no history of alcoholism, hazard ratio (HR): 1.43, 95% confidence interval (CI): 0.88, 2.34; history of alcoholism, HR: 0.71, 95% CI: 0.40, 1.28). Ensuring timely access to HIV care remains a challenge in the southeastern USA. Affectionate, tangible, and emotional/informational social support were not associated with the time from diagnosis to care. The presence of positive social interaction may be an important factor influencing care-seeking behavior after diagnosis. PMID- 20024775 TI - Cost of OraQuick oral fluid rapid HIV testing at 35 community clinics and community-based organizations in the USA. AB - The US Centers for Disease Control and Prevention recommends the expanded use of rapid HIV antibody tests in community settings to increase the proportion of persons who are aware of their HIV status. The cost of providing rapid testing services in these settings is not known. We conducted a cost survey of non-profit community clinics and community-based organizations (CBOs). The final study sample for the cost analyses included 28 community clinics and seven CBOs that offered rapid HIV testing, all of which used OraSure's OraQuick oral fluid assay. Overall, the mean per-client rapid testing cost was 36.68 dollars for HIV negative clients (2006 dollars, provider perspective) and 44.22 dollars for preliminary-positive clients. Personnel costs accounted for 55.6% of overall testing costs for negative clients and 63.2% for preliminary-positive clients. This study contributes unique empirical data about the costs of OralQuick rapid testing that can be used by clinic and CBO directors to assess the economic impact of implementing rapid testing at their sites. The findings also could be used to inform discussions regarding national and local HIV testing policies. PMID- 20024776 TI - Validity of the K-10 in detecting DSM-IV-defined depression and anxiety disorders among HIV-infected individuals. AB - It has been suggested that an HIV diagnosis may increase the likelihood of mental disorders among infected individuals and that the progression of HIV may be hastened by mental disorders like anxiety and depression. Therefore, a brief screening measure, with good sensitivity/specificity for psychiatric diagnoses that could be given to HIV-infected individuals would be useful. We assessed the validity of the K-10, using the MINI International Neuropsychiatric Interview as the gold standard, in a sample of 429 HIV-infected adults enrolled in HIV care and treatment services near Cape Town, South Africa. There was significant agreement between the K-10 and the MINI-defined depressive and anxiety disorders. A receiver operating characteristic (ROC) curve analysis indicated that the K-10 showed agreeable sensitivity and specificity in detecting depression (area under the ROC curve, 0.77), generalized anxiety disorder (0.78), and posttraumatic stress disorder (PTSD) (0.77). The K-10 may be a useful screening measure for detecting mood and anxiety disorders, including PTSD, in patients with HIV/AIDS. PMID- 20024777 TI - The role of minor and adult children in the adjustment of women living with HIV. AB - The present study examined how having children can relate to the psychological functioning of HIV-positive women, and the place and function children have in their mother's social support and social burden networks. As part of initial face to-face interviews with 46 HIV-positive women enrolled in a longitudinal study, participants indicated their maternal status, the nature of their social support and burden (with a modified Multidimensional Social Support Inventory) and their levels of depressive symptoms (Brief Symptom Inventory). Using Analysis of Variance (ANOVA) and non-parametric analyses when statistical assumptions were not met, findings indicate that women with minor children reported fewer depressive symptoms and more confidence in regulating social support than women without children. Maternal status was not significantly related to other measures of social support. Women with minor children reported greater comfort and confidence in regulating social burden and reported experiencing less social burden than women with adult children only. Women with adult children only reported intermediate levels of depression, but high levels of received burden and low levels of comfort and confidence in regulating (lessening) this burden. Analyses of women's social networks (Fisher's Exact Test) indicated that minor children were less likely to be named regarding support than adult or other family members and were infrequently turned to for practical, emotional, or HIV related support. Adult children were more often listed as providing various types of support than minor children, yet these relationships were also often associated with relatively higher levels of social burden. PMID- 20024778 TI - Methamphetamine initiation among HIV-positive gay and bisexual men. AB - This study describes factors associated with methamphetamine initiation in a racially diverse sample of 340 methamphetamine-using, HIV-positive gay and bisexual men. A factor analysis was conducted on reasons for initiation, and four factors were identified: to party, to cope, for energy, and to improve self esteem. Methamphetamine to party accounted for more than one-third of the variance in the factor analysis. Methamphetamine to cope captured almost 9% of the variance, methamphetamine for energy accounted for approximately 8% of the variance, and methamphetamine for self-esteem accounted for approximately 7% of the variance. Regression analyses revealed differential associations between methamphetamine-initiation factors and HIV-risk behaviors. Methamphetamine for self-esteem predicted binge methamphetamine use, while methamphetamine to cope was associated with injecting methamphetamine. Using methamphetamine for energy was associated with number of illicit drugs-used and using methamphetamine to party was associated with having a greater number of sexually transmitted infections. These findings suggest that methamphetamine initiation among gay and bisexual men is multifaceted, which could have implications for intervention development. PMID- 20024779 TI - Anal use of the female condom: does uncertainty justify provider inaction? AB - Despite limited safety data and the absence of efficacy data, several studies have reported that the female condom is being used for anal sex by men who have sex with men. We describe providers' awareness of female condom use during anal sex among their clients and their experiences in counseling clients. We conducted semi-structured interviews with 78 health-care providers recruited from various health-care delivery systems in New York City: a family planning agency, a sexually transmitted infection agency, a hospital-based obstetrics and gynecology clinic, and two community-based AIDS service organizations. While two-thirds of providers reported that they were uncertain as to whether the female condom could or should be used for anal intercourse, nearly one-third believed that anything is better than nothing to prevent HIV/sexually transmitted infections during anal sex. Few providers had actually talked with clients about anal use of the female condom, and clients themselves had seldom mentioned nor asked for information about such use. Our findings highlight providers' uncertainty about anal use of the female condom. Lacking guidelines regarding the safety and efficacy of female condom use during anal sex, health-care providers are left to make their own well intentioned recommendations (or not) to potential users. The dearth of information on female condom use during anal sex could encourage individuals to use the female condom for anal sex, which may increase HIV transmission risk or represent a missed opportunity for protecting non-condom users. There is a need for a series of harm-reduction, acceptability, and efficacy studies and, in the interim, for the development of a carefully qualified safety set of guidelines regarding anal use of the female condom for health-care providers. PMID- 20024780 TI - Efficacy of convenience sampling through the internet versus respondent driven sampling among males who have sex with males in Tallinn and Harju County, Estonia: challenges reaching a hidden population. AB - This paper examines challenges obtaining representative samples of males who have sex with males (MSM) in Estonia and provides descriptive HIV behavioral data gathered from four cross-sectional surveys; three using the internet, and one using respondent driven sampling (RDS) to recruit MSM in Tallinn and Harju County. Estonian MSM were sampled between March and May in 2004 (n=193), August and November in 2005 (n=146) and September and December in 2007 (n=238) using internet websites. MSM in Tallinn and Harju County were sampled between April and June in 2007 (n=59) using RDS. Recruitment of MSM using RDS did not acquire the calculated sample size. The RDS study reached a less diverse group of MSM than did the internet studies which recruited a larger proportion of MSM who were older, bisexual, having female sexual partners during the last six months, and unlikely to have been tested for HIV. The findings and observations presented in this paper could inform researchers in Estonia, and the region, about the efficacy of and socio-cultural challenges to sampling MSM to collect HIV biological and/or behavioral data. PMID- 20024781 TI - Factors associated with late HIV testing for Latinos diagnosed with AIDS in Los Angeles. AB - Latinos are more likely to test late for HIV infection compared to other racial/ethnic groups in the United States. A population-based interview study was used to examine factors associated with late HIV testing for Latinos diagnosed with AIDS in Los Angeles County (LAC) to develop more effective HIV testing outreach strategies. Latinos testing for HIV within one year of an AIDS diagnosis were considered as late testers, while those diagnosed with AIDS more than one year after an HIV diagnosis were defined as non-late testers. After adjusting for age, education, country of birth, and injection drug use in a logistic regression analysis, completion of the interview in Spanish was the main factor associated with late testing (adjusted Odds Ratio (AOR) = 2.9, 95% Confidence Intervals (CIs): 1.4, 6.0). Latinos testing late for HIV were also more likely to test due to illness (p<0.0001) and less likely to test as part of a clinical screening (p<0.0001). Late testers were more likely to receive their first positive HIV test as a hospital inpatient (p<0.0001) and less likely to test positive at a community health center or public clinic (p=0.05). To accomplish widespread and timely HIV testing for Latinos in LAC, Spanish-language social marketing campaigns are needed and Spanish-speaking patients should be offered HIV testing in all clinical settings. PMID- 20024782 TI - Effect of scrapie on the stability of housekeeping genes. AB - Scrapie is the archetype of prion diseases, fatal neurodegenerative disorders that affect humans and animals. Gene expression analysis of normal and infected sheep may provide clues to clarify the molecular mechanisms involved in the neuropathology of these diseases. Real time quantitative PCR has become a powerful and accurate technique for examination of transcription patterns in different biological conditions. One of the critical steps in the comparison of transcription profiles is the selection of stable genes for normalization of expression data. In this work, we have investigated the effect of scrapie on the stability of eight commonly used housekeeping genes in the central nervous system of sheep. We found that their stability decreased in scrapie-infected tissues, with the effect of the disease most evident in the medulla oblongata, a highly affected area of the brain stem. The risk of choosing inappropriate housekeeping genes for expression analysis was evaluated. Although the stability of each reference gene was suitable, a wide variation in expression of target genes (BAX and BCL2) was observed when only one or two housekeeping genes were used to normalize. However, reliable results were obtained with a normalization factor based on three reference genes, regardless of their position in a stability ranking. PMID- 20024783 TI - Associations of A-FABP and H-FABP markers with the content of intramuscular fat in Beijing-You chicken. AB - This study has assessed the association of single nucleotide polymorphisms (SNP) identified in the adipocyte fatty acid binding protein (A-FABP) and heart-type fatty acid binding protein (H-FABP) genes with the content of intramuscular fat (IMF) in a population of male Beijing-You chickens. A previously described SNP in the chicken A-FABP gene had a significant (P < 0.05) effect on IMF content. Chickens inheriting the homozygous BB genotype at A-FABP had a significantly higher content of IMF in thigh muscles and breast muscles than did those inheriting the AA and AB genotypes. A novel SNP, identified here, in the H-FABP gene was also significantly (P < 0.05) associated with IMF content in thigh and breast muscle. Chickens inheriting the genotypes of DD and CD had much higher content of IMF than those inheriting the homozygous genotype of CC. Markers at the A-FABP and H-FABP genes were associated with IMF content in the studied population. Chickens inheriting the BB genotype at A-FABP, along with the CD genotype at H-FABP, produced muscles with a much higher content of IMF when compared with all other genotypes. A weak interaction between A-FABP and H-FABP was detected (P < 0.09) for IMF content in the tested population. The statistical significance of interaction is tentative because of the limited number of observations for some genotypic combinations. Markers identified within the A FABP and H-FABP genes are suitable for future use in identifying chickens with the genetic potential to produce more desirable muscle with higher IMF content, at least in the population of Beijing-You male chickens. PMID- 20024784 TI - Yak (Bos grunniens) stomach lysozyme: molecular cloning, expression and its antibacterial activities. AB - The cDNA coding for stomach lysozyme in yak was cloned. The cloned cDNA contains a 432 bp open reading frame and encodes 143 amino acids (16.24 KDa) with a signal peptide of 18 amino acids. Further analysis revealed that its amino acid sequence shares many common properties with cow milk lysozyme. Expression of this gene was also detected in mammary gland tissue by RT-PCR. Phylogenetic relationships among yak stomach lysozyme and 8 cow lysozymes indicated that the yak enzyme is more closely related to both cow milk lysozyme and the pseudogene PsiNS4 than cow stomach lysozyme. Recombinant yak lysozyme purified by Ni(2+)-column showed a molecular weight of 33.78 kDa and exhibited lytic activity against Staphylococcus aureus, providing evidence of its antibacterial activities. PMID- 20024785 TI - A single nucleotide polymorphism and sequence analysis of CSN1S1 gene promoter region in Chinese Bos grunniens (yak). AB - The aim of this study was to investigate the polymorphism of the CSN1S1 gene promoter region in 4 Chinese yak breeds, and compare the yak CSN1S1 gene promoter region sequences with other ruminants. A Polymerase Chain Reaction-Single Strand Conformation Polymorphism protocol was developed for rapid genotyping of the yak CSN1S1 gene. One hundred fifty-eight animals from 4 Chinese yak breeds were genotyped at the CSN1S1 locus using the protocol developed. A single nucleotide polymorphism of the CSN1S1 gene promoter region has been identified in all yak breeds investigated. The polymorphism consists of a single nucleotide substitution G-->A at position 386 of the CSN1S1 gene promoter region, resulting in two alleles named, respectively, G(386) and A(386), based on the nucleotide at position 386. The allele G(386) was found to be more common in the animals investigated. The corresponding nucleotide sequences in GenBank of yak (having the same nucleotides as allele G(386) in this study), bovine, water buffalo, sheep, and goat had similarity of 99.68%, 99.35%, 97.42%, 95.14%, and 94.19%, respectively, with the yak allele A(386.). PMID- 20024786 TI - A single nucleotide polymorphism of chicken acetyl-CoA carboxylase A gene associated with fatness traits. AB - Acetyl-CoA carboxylase alpha (ACCalpha) is a major rate-limiting enzyme in the biogenesis of long-chain fatty acids. It can catalyze the carboxylation of acetyl CoA to form malonyl-CoA that plays a key role in the regulation of fatty acid metabolism. The objective of the present study was to investigate the associations of ACCalpha gene polymorphisms with chicken growth and body composition traits. The Northeast Agricultural University broiler lines divergently selected for abdominal fat content and the Northeast Agricultural University F(2) Resource Population were used in the current study. Body weight and body composition traits were measured in the aforementioned two populations. A synonymous mutation was detected in the exon 19 region of ACCalpha gene, then polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method was developed to genotype all the individuals derived from the aforementioned populations. Association analysis revealed that the polymorphism was associated with abdominal fat weight and percentage of abdominal fat in the two populations. The results suggested that ACCalpha gene could be a candidate locus or linked to a major gene that affects abdominal fat content in the chicken. PMID- 20024787 TI - Expression studies of the obesity candidate gene FTO in pig. AB - Obesity is an increasing problem worldwide and research on candidate genes in good animal models is highly needed. The pig is an excellent model as its metabolism, organ size, and eating habits resemble that of humans. The present study is focused on the characterization of the fat mass and obesity associated gene (FTO) in pig. This gene has recently been associated with increased body mass index in several human populations. To establish information on the expression profile of FTO in the pig we performed quantitative PCR in a panel of adult pig tissues and in tissues sampled at different developmental stages. Expression of the FTO transcript was detected in all tissues tested with significantly higher levels in brain tissues (cortex, cerebellum, and hippocampus; P < 0.001). These levels varied through the development and between the specific parts of the brain studied (i.e., frontal cortex and cerebellum). Additionally, in order to see the involvement of the FTO gene in obesity, the changes in expression level were investigated in a nutritional study in brain of Gottingen minipigs under a high cholesterol diet. Significantly higher (P < 0.01) levels of FTO transcript were found in cortex when compared with cerebellum of the high-cholesterol fed pigs. Furthermore, SNPs were investigated in the coding sequence of the FTO in the Gottingen minipig and in the Danish commercial pig. Eleven synonymous SNPs and a two bp insertion were found between the two pig lines. PMID- 20024788 TI - Molecular cloning and comparative characterization of the porcine troponin I family. AB - Troponin I (TnI) is a family of three muscle-specific myofibrillar proteins involved in calcium-sensitive regulation of contraction in cardiac and skeletal muscle. In this study, the full-length cDNA and genomic sequence of three genes of porcine TnI family were cloned and sequenced. The full-length cDNA of TNNI1, TNNI2, and TNNI3 genes were 989 bp, 734 bp, and 831 bp in length, which contained an open reading frame of 564, 549, and 636 nucleotides, respectively. Three Troponin I shared 54.4 approximately 58.3% similarity with each other in their predicted amino acid sequences. The TNNI1, TNNI2, and TNNI3 displayed the same genomic structure as other vertebrates and spanned over 9785 bp, 2373 bp, and 3648 bp genomic regions, respectively. The regulatory elements in the proximal promoter of TNNI2 and TNNI3 were conserved among human, mouse, and pig, but regulatory element differences existed in the TNNI1 promoter among them. Expression profiling showed that TnI genes were widely expressed in the tissues studied, with the highest expression level of TNNI1 and TNNI2 in skeletal muscle, and TNNI3 in cardiac muscle. PMID- 20024789 TI - Biofouling control in water by various UVC wavelengths and doses. AB - UV light irradiation is being increasingly applied as a primary process for water disinfection, effectively used for inactivation of suspended (planktonic) cells. In this study, the use of UV irradiation was evaluated as a pretreatment strategy to control biofouling. The objective of this research was to elucidate the relative effectiveness of various targeted UV wavelengths and a polychromatic spectrum on bacterial inactivation and biofilm control. In a model system using Pseudomonas aeruginosa, the inactivation spectra corresponded to the DNA absorption spectra for all wavelengths between 220 and 280 nm, while wavelengths between 254 nm and 270 nm were the most effective for bacterial inactivation. Similar wavelengths of 254-260-270 nm were also more effective for biofilm control in most cases than targeted 239 and 280 nm. In addition, the prevention of biofilm formation by P. aeruginosa with a full polychromatic lamp was UV dose dependent. It appears that biofilm control is improved when larger UV doses are given, while higher levels of inactivation are obtained when using a full polychromatic MP lamp. However, no significant differences were found between biofilms produced by bacteria that survived UV irradiation and biofilms produced by control bacteria at the same microbial counts. Moreover, the experiments showed that biofilm prevention depends on the post-treatment incubation time and nutrient availability, in addition to targeted wavelengths, UV spectrum and UV dose. PMID- 20024790 TI - O-glycosylation expression in fibroadenoma. AB - Fibroadenomas are human benign breast tumors characterized by proliferation of epithelial and stroma cells of the terminal ductal unit. Expression of O-glycans seems to contribute to the proliferation and transformation events. With this in mind, we evaluated the expression of glycans in fibroadenoma tissue through immunohistochemistry with antibodies against mucin epitopes (Anti CA15-3 and MUC1), as well as with lectins specific for glycans linked to proteins or lipids, and we compared findings with healthy breast specimens. Our results show positive expression of CA15-3 and MUC1 in fibroadenoma tissue, mainly in duct and stroma cells, whereas, in normal samples, staining was observed in duct cells. The lectin from Glycine max recognized equally well duct and stroma cells; this was the only lectin showing co-localization with anti-CA15-3 in healthy and tumor tissues. Dolichos biflorus, Artocarpus integrifolia, and Griffonia simplicifolia lectins recognized duct cells in control healthy tissues as well as in fibroadenoma tissue. The lectin from Amaranthus leucocarpus recognized only duct cells in control samples, whereas, in fibroadenoma tissue, it recognized duct and some stromal cells, suggesting that O-glycans-type mucin linked to proteins and mucin participate in the development of fibroadenomas. PMID- 20024791 TI - A large scale method for preparation of plant thylakoids for use in body weight regulation. AB - A method for preparation of thylakoids from plant leaves on a large scale is described. The method involves: 1) disruption of the cells with a blender followed by filtration to remove large cell debris and non disrupted cells. 2) precipitation of the thylakoids by adjusting the pH to the isoelectric point, pH 4.7. 3) a washing step by dilution of the precipitate in water followed by precipitation at the same pH. 4) concentration of the precipitate by freeze- thawing or freeze -drying to get the final product. The product is characterized, with respect to protein composition, by SDS-PAGE and mass-spectroscopy, the content of carotenoids, particularly the xanthophylls violaxanthin, antheraxanthin, and zeaxanthin. The thylakoid preparation has about the same capacity to inhibit pancreatic lipase/colipase activity as thylakoids prepared by standard laboratory methods using sucrose in the medium and centrifugation. In a study with mice, it was found that, when the thylakoids were added to the food over 32 days, they significantly reduced the body weight gain and the percentage body fat. The large scale method described here allows studies on the effect of thylakoids in appetite regulation on experimental animals in a longer lasting time and also on humans. PMID- 20024792 TI - Extracellular lipolytic enzyme activity of a newly isolated Debaryomyces hansenii. AB - A strain isolated from waste of a milk products plant and exhibited extracellular lipolytic activity was identified as Debaryomyces hansenii by 5.8S rRNA and 28S rRNA gene sequence analyses. Lipolytic activity was assayed spectrophotometrically by using p-nitrophenylpalmitate. Higher specific lipolytic activities were obtained in the presence of tristearin (0.68 U/mg prot), oleic acid (0.56 U/mg prot), and soybean oil (0.36 U/mg prot) than other triglycerides, fatty acids, and vegetable oils considered as carbon sources. Cheese whey appeared to be a good alternative to lipidic substances for lipolytic activity. Among various organic and inorganic nitrogen sources, soy flour was found to attain the lipolytic activity similar to that provided by universal yeast medium components. This work is the first report on the discussion of lipolytic activity enhancement by D. hansenii through modulating the cultivation medium. It also proposes low cost medium nutrients that could be of industrial value and could serve as basal nutrients for further optimization studies on the lipase production by D. hansenii. PMID- 20024793 TI - Enhanced production of 6-aminopenicillanic acid in aqueous methyl isobutyl ketone system with immobilized penicillin G acylase. AB - Enzymatic hydrolysis of penicillin G for production of 6-amino-penicillanic-acid (6-APA) was achieved by using penicillin G acylase as catalyst in an aqueous methylisobutyl ketone (MIBK) system. The optimization was carried out and it was found that the best conversion was improved 10% more than the aqueous system, which was obtained at the conditions: initial pH 8.0, 5.0% (W/V) substrate (penicillin G), and temperature at 35 degrees C, and the ratio of aqueous and organic phase was 3:1. The stability of the biocatalyst was studied at the operational conditions. After 5 cycles of semi-batch reactions, the residual activity of penicillin G acylase was 69.2% of the initial activity. There was no apparent loss of the yield of product. This process has a potential application in the industrial scale production of 6-APA because it simplifies the process effectively. PMID- 20024794 TI - Construction and expression of a spliced variant of Epstein-Barr virus bzlf1 and preparation of its polyclonal antibody. AB - The BZLF1 gene-encoded protein, Zta (EB1, ZEBRA), is a key transcriptional activator of induction of the lytic cycle of EBV. Zta; it contains a basic region with homology to the DNA binding domains of the AP-1 family. In this study, an alternatively spliced BZLF1 (Delta BZLF1) cDNA lacking exon 2, which encodes the DNA-binding domain of Zta, was isolated from B95-8 marmoset cell line releasing EBV. The cDNA was inserted into a prokaryotic expression vector pET-28a+. The His tagged recombinant protein was overproduced in E. coli BL21(DE3) and purified by nickel affinity chromatography. The purified fraction was characterized by Western blot and MALDI-TOF-MS analysis and used as an antigen to immunize mice. The antibody against Delta Zta can recognize both denatured and natural Zta protein. The Delta Zta protein and its antibody can be used to further investigate its unknown functions. PMID- 20024795 TI - Preparation and activity of bubbling-immobilized cellobiase within chitosan alginate composite. AB - Cellobiase can hydrolyze cellobiose into glucose; it plays a key role in the process of cellulose hydrolysis by reducing the product inhibition. To reuse the enzyme and improve the economic value of cellulosic ethanol, cellobiase was immobilized using sodium alginate and chitosan as carriers by the bubbling method. The immobilization conditions were optimized as follows: enzyme loading of 100 U cellobiase/g carrier, 30 min immobilization, 3.5 wt% sodium alginate, 0.25 wt% chitosan, and 2 wt% calcium chloride. Compared to free enzyme, the immobilized cellobiase had a decreased apparent K(m) and the maximum activity at a lower pH, indicating its higher acidic and thermal stability. The immobilized cellobiase was further tested in the hydrolysis of cellobiose and various cellulosic substrates (microcrystalline cellulose, filter paper, and ammonia pretreated corn cobs). Together with cellulases, the immobilized cellobiase converted the cellulosic substrates into glucose with the rate and extent similar to the free enzyme. PMID- 20024796 TI - Enhanced stability of Bacillus licheniformis L-arabinose isomerase by immobilization with alginate. AB - Recombinant Escherichia coli whole cells harboring Bacillus licheniformis L arabinose isomerase (BLAI) were harvested to prepare alginate-immobilized biocatalysts. The operational conditions for immobilization were optimized according to relative activity and the cell leakage of the immobilized cell. The optimal conditions are as follows: alginate concentration, Ca(2+) concentration, cell mass loading, and curing time were 2% (w/v), 0.1 M, 50 g l(-1), and 4 hours, respectively. After immobilization, cross-linking with 0.1% glutaraldehyde significantly reduced cell leakage. The immobilized whole cells harboring BLAI were very stable with 89% residual activity remaining after 33 days of incubation at 50 degrees C and were much more stable than the free enzyme and cells. The results showed that immobilizing whole cells harboring BLAI is suitable for use as a biocatalyst in the production of L-ribulose, largely due to its high stability and low cost. PMID- 20024797 TI - Identification of carotenoids produced from cheese whey by Blakeslea trispora in submerged fermentation. AB - The identification of carotenoids in B. trispora during pigment production from deproteinized hydrolyzed whey supplemented with plant oils was studied. The carotenoid content in Blakeslea trispora were beta-carotene, gamma-carotene, and lycopene. The composition of carotenoids depends of the amount of oils added to the cheese whey. At the maximum concentration of carotenoids, the proportions of beta-carotene, gamma-carotene, and lycopene (as percent of total carotenoids) was 60.1%, 32.5%, and 7.4%, respectively. PMID- 20024798 TI - Optimum pyrolytic conditions for furan derivative formation from polyamino sugars. AB - Furan derivatives were prepared by hydrolysis of glucosamine derived from polyamino sugars assisted with pyrolysis in alkaline condition. Pyrolysis was carried out with autoclaving at 15 psi/121 degrees C/1 hour. The reaction was monitored by observing spectral characteristics at lambda(max) 545 nm with p dimethylaminobenzaldehyde. Alkaline hydrolysis favors the formation of furan derivatives in N-acetylglucosamine, glucosamine, hyaluronic acid, but not in dermatan sulfate and chondroitin sulfate. These results indicate -OH group orientation in the cis position at C(4) of the Haworth projection formula of hexosamines may be playing a role in the formation of furans. This can be a useful method for estimation of the glucosamine in glycoprotein and urine. PMID- 20024799 TI - Conversion of trypsin to a copper enzyme: tyrosinase/catechol oxidase by chemical modification. AB - New active sites can be introduced into naturally occurring enzymes by the chemical modification of specific amino acid residues in concert with genetic techniques. Chemical strategies have had a significant impact in the field of enzyme design such as modifying the selectivity and catalytic activity which is very different from those of the corresponding native enzymes. Thus, chemical modification has been exploited for the incorporation of active site binding analogs onto protein templates and for atom replacement in order to generate new functionality such as the conversion of a hydrolase into a peroxidase. The introduction of a coordination complex into a substrate binding pocket of trypsin could probably also be extended to various enzymes of significant therapeutic and biotechnological importance. The aim of this study is the conversion of trypsin into a copper enzyme: tyrosinase by chemical modification. Tyrosinase is a biocatalyst (EC.1.14.18.1) containing two atoms of copper per active site with monooxygenase activity. The active site of trypsin (EC 3.4.21.4), a serine protease was chemically modified by copper (Cu(+2)) introduced p-aminobenzamidine (pABA- Cu(+2): guanidine containing schiff base metal chelate) which exhibits affinity for the carboxylate group in the active site as trypsin-like inhibitor. Trypsin and the resultant semisynthetic enzyme preparation was analysed by means of its trypsin and catechol oxidase/tyrosinase activity. After chemical modification, trypsin-pABA-Cu(+2) preparation lost 63% of its trypsin activity and gained tyrosinase/catechol oxidase activity. The kinetic properties (K(cat), K(m), K(cat)/K(m)), optimum pH and temperature of the trypsin-pABA-Cu(+2) complex was also investigated. PMID- 20024801 TI - Molecular dynamics study of prolyl oligopeptidase with inhibitor in binding cavity. AB - We used the crystal structure of prolyl oligopeptidase (POP) with bound Z-pro prolinal (ZPP) inhibitor (Protein Data Bank (PDB) structure 1QFS) to perform an intensive molecular dynamics study of the POP-ZPP complex. We performed 100 ns of simulation with the hemiacetal bond, through which the ZPP is bound to the POP, removed in order to better investigate the binding cavity environment. From basic analysis, measuring the radius of gyration, root mean square deviation, solvent accessible surface area and definition of the secondary structure of protein, we determined that the protein structure is highly stable and maintains its structure over the entire simulation time. This demonstrates that such long time simulations can be performed without the protein structure losing stability. We found that water bridges and hydrogen bonds play a negligible role in binding the ZPP thus indicating the importance of the hemiacetal bond. The two domains of the protein are bound by a set of approximately 12 hydrogen bonds, specific to the particular POP protein. PMID- 20024802 TI - The acid-base profile of a contemporary set of drugs: implications for drug discovery. AB - Acid-base ionization constant (pK(a)) values have considerable influence on the physicochemical and pharmacokinetic properties of therapeutic substances. A set of 907 drugs was examined to determine the proportion of drugs that contain an ionizable group and the distribution of their pK(a) values. Using this contemporary set of compounds it was found that 64% of these compounds contained an ionizable group. Within this group of ionizable compounds, 34% contained a single basic group while only 20% contained a single acidic functional group. The single acid and single base containing substances were investigated further to examine the distributions of their pK(a) values. These data are discussed and analyzed with a focus on the entire set as well as central nervous system, non central nervous system and oral drugs. The findings from this research will prompt pharmaceutical companies to assess the constitution of their screening libraries, such that focus is placed on the proportion of ionizable substances, the ratio of acids to bases and the distribution of pK(a) values. PMID- 20024803 TI - Mechanism-based categorization of aromatase inhibitors: a potential discovery and screening tool. AB - Cytochrome P450 aromatase is a key steroidogenic enzyme that converts androgens to estrogens in vertebrates. There is much interest in aromatase inhibitors (AIs) both because of their use as pharmaceuticals in the treatment of estrogen sensitive breast cancers, and because a number of environmental contaminants can act as AIs, thereby disrupting endocrine function in humans and wildlife through suppression of circulating estrogen levels. The goal of the current work was to develop a mechanism-based structure-activity relationship (SAR) categorization framework highlighting the most important chemical structural features responsible for inhibition of aromatase activity. Two main interaction mechanisms were discerned: steroidal and non-steroidal. The steroid scaffold is most prominent when the structure of the target chemical is similar to the natural substrates of aromatase - androstenedione and testosterone. Chemicals acting by non-steroidal mechanism(s) possess a heteroatom (N, O, S) able to coordinate the heme iron of the cytochrome P450, and thus interfere with steroid hydroxylation. The specific structural boundaries controlling AI for both analyzed mechanisms were defined, and a software tool was developed that allowed a decision tree (profile) to be built discriminating AIs by mechanism and potency. An input chemical follows a profiling path and the structure is examined at each step to decide whether it conforms with the structural boundaries implemented in the decision tree node. Such a system would aid drug discovery efforts, as well as provide a screening tool to detect environmental contaminants that could act as AIs. PMID- 20024804 TI - QNA-based 'Star Track' QSAR approach. AB - In the existing quantitative structure-activity relationship (QSAR) methods any molecule is represented as a single point in a many-dimensional space of molecular descriptors. We propose a new QSAR approach based on Quantitative Neighbourhoods of Atoms (QNA) descriptors, which characterize each atom of a molecule and depend on the whole molecule structure. In the 'Star Track' methodology any molecule is represented as a set of points in a two-dimensional space of QNA descriptors. With our new method the estimate of the target property of a chemical compound is calculated as the average value of the function of QNA descriptors in the points of the atoms of a molecule in QNA descriptor space. Substantially, we propose the use of only two descriptors rather than more than 3000 molecular descriptors that apply in the QSAR method. On the basis of this approach we have developed the computer program GUSAR and compared it with several widely used QSAR methods including CoMFA, CoMSIA, Golpe/GRID, HQSAR and others, using ten data sets representing various chemical series and diverse types of biological activity. We show that in the majority of cases the accuracy and predictivity of GUSAR models appears to be better than those for the reference QSAR methods. High predictive ability and robustness of GUSAR are also shown in the leave-20%-out cross-validation procedure. PMID- 20024805 TI - Assessing the reproductive toxicity of some (con)azole compounds using a structure-activity relationship approach. AB - The present research investigates the study of a set of 27 (con)azoles and their reproductive toxicity. (Con)azoles are used as fungicides and herbicides in agriculture for treatment of fruits, vegetables, cereals, and seeds, or as human antimycotic therapeutics. According to EEC Directive 91/414, active substances used in plant protection products must undergo reproductive toxicity testing. Reproductive toxicity is a complex biological endpoint, which includes many different biological processes and, therefore, it can only to a limited extent be assessed by a single quantitative structure-activity relationship (QSAR) model. The proposed SAR models are built using unsupervised methods, such as hierarchical clustering, principal component analysis and self-organizing maps, with the aim of studying the similarity relationships between structures. The molecular structures are represented with a set of topological and structural descriptors. The models showing clusters, closest neighbours or outliers may support the categorization and the classification of (con)azoles as potential reproductive toxicants. PMID- 20024806 TI - Modelling the relative toxicity of metals on respiration of nitrifiers using ion characteristics. AB - The effects of eight transition metals were studied in a nitrifying system to investigate the relationship between the ionic characteristics of metals and their toxicity to nitrifiers. The cumulative oxygen consumption and the cumulative carbon dioxide production were monitored throughout each respirometric batch run to determine the toxicity of metals to nitrifiers. Several quantitative cationic-activity relationship (QCAR) models were developed on the basis of these different toxicity endpoints using quantum chemical descriptors. Descriptors were calculated with density functional theory (DFT) at the B3LYP/LANL2DZ level using the Gaussian 03W software. Additionally, the same descriptors were recalculated using another basis set, B3LYP/SDD, to test the impact of the basis set on prediction of toxicity. Of the calculated descriptors, mainly the gaseous phase descriptors explained significant variances in both toxicity endpoints. However, the energy of the polarized solute-solvent (E(PSS)) was the most important common descriptors in modelling labile toxicity. A combination of the aqueous phase energy of the highest occupied molecular orbital (E(HOMO(aq))) and the maximum value for the energy of the lowest unoccupied molecular orbital of the most important metal species (E(LUMO)C(max)) produced the best two-descriptor model for both pTO(2) and pTCO(2). The electron donor/acceptor ability of metals and the electron acceptor ability of metal species (E(LUMO)C(max)) seemed to be important in explaining toxicity in aqueous media regardless of the measured endpoints for nitrifiers. PMID- 20024807 TI - Data-driven model for the prediction of protein transmembrane regions. AB - We present a novel approach combining mathematical methods and artificial neural networks to predict the transmembrane regions of transmembrane proteins, considering protein sequence information alone. We have focused on developing a data-driven model based on a non-linear modelling method, the counter-propagation artificial neural network, and on mathematical descriptors defining the sequence information of transmembrane proteins with known three-dimensional structures. The developed model has proven to be promising in predicting protein transmembrane regions, with an error below 10% for the external validation set. In combination with available experimental data the model can give us a better understanding of transmembrane proteins. PMID- 20024808 TI - In silico method for identification of promising anticancer drug targets. AB - In recent years, the accumulation of the genomics, proteomics, transcriptomics data for topological and functional organization of regulatory networks in a cell has provided the possibility of identifying the potential targets involved in pathological processes and of selecting the most promising targets for future drug development. We propose an approach for anticancer drug target identification, which, using microarray data, allows discrete modelling of regulatory network behaviour. The effect of drugs inhibiting a particular protein or a combination of proteins in a regulatory network is analysed by simulation of a blockade of single nodes or their combinations. The method was applied to the four groups of breast cancer, HER2/neu-positive breast carcinomas, ductal carcinoma, invasive ductal carcinoma and/or a nodal metastasis, and to generalized breast cancer. As a result, some promising specific molecular targets and their combinations were identified. Inhibitors of some identified targets are known as potential drugs for therapy of malignant diseases; for some other targets we identified hits in the commercially available sample databases. PMID- 20024809 TI - Quantitative structure-activity relationship modelling of oral acute toxicity and cytotoxic activity of fragrance materials in rodents. AB - Fragrance materials are used as ingredients in many consumer and personal care products. The wide and daily use of these substances, as well as their mainly uncontrolled discharge through domestic sewage, make fragrance materials both potential indoor and outdoor air pollutants which are also connected to possible toxic effects on humans (asthma, allergies, headaches). Unfortunately, little is known about the environmental fate and toxicity of these substances. However, the use of alternative, predictive approaches, such as quantitative structure activity relationships (QSARs), can help in filling the data gap and in the characterization of the environmental and toxicological profile of these substances. In the proposed study, ordinary least squares regression-based QSAR models were developed for three toxicological endpoints: mouse oral LD(50), inhibition of NADH-oxidase (EC(50) NADH-Ox) and the effect on mitochondrial membrane potential (EC(50) DeltaPsim). Theoretical molecular descriptors were calculated by using DRAGON software, and the best QSAR models were developed according to the principles defined by the Organization for Economic Co-operation and Development. PMID- 20024814 TI - Separate but unequal: addressing disparities through health care reform. PMID- 20024815 TI - The Intercultural Cancer Council's Fact Sheet series: a tool for public and professional education. PMID- 20024816 TI - Native People's Circle of Hope. PMID- 20024817 TI - Leadership in public health. PMID- 20024818 TI - The need for justice in human participants research: life after the Belmont Commission. PMID- 20024819 TI - Making cultural competence work for cancer prevention: fact of fiction. PMID- 20024820 TI - Shaping oncology care for the future: an Oncology Nursing Society perspective. PMID- 20024821 TI - Issues, priorities, and political implications for cancer: an organization's perspective. PMID- 20024822 TI - Using a biopsychosocial approach to address health disparities--one person's vision. PMID- 20024823 TI - Programming for results. PMID- 20024824 TI - The case for Eliminating Disparities in Clinical Trials. PMID- 20024825 TI - Policies to address disparities in clinical trials: the EDICT Project. PMID- 20024826 TI - Implementing recommendations to eliminate disparities in clinical trial research. PMID- 20024827 TI - EDICT policy recommendations support protection of research participants. PMID- 20024828 TI - Addressing disparities in clinical trials: Culturally and Linguistically Appropriate Standards in Clinical Trials (CLAS-ACT) and the EDICT BackPack initiative. PMID- 20024829 TI - Making the invisible visible: professional education to eliminate disparities in clinical trials. PMID- 20024830 TI - How Medicare supports participation of beneficiaries in clinical trials. PMID- 20024831 TI - EDICT and the Intercultural Cancer Council. PMID- 20024832 TI - Breast health issues of undocumented women in California and Texas. PMID- 20024833 TI - Eyes on the Pacific: cancer issues of Native Hawaiians and Pacific Islanders in Hawai'i and the US-associated Pacific. PMID- 20024834 TI - Collegiate Cancer Council: University of Houston Chapter. PMID- 20024835 TI - From cancer alley to biotech: the importance of mentors. PMID- 20024836 TI - American Heart Association--Scientific Sessions 2009. Part 1. AB - The American Heart Association (AHA) 2009 meeting held in Orlando, FL, USA included topics covering new developments in the field of cardiovascular disease. This conference report highlights selected presentations on translational science, cardiovascular disease genomics and the US healthcare reform. PMID- 20024837 TI - American Heart Association--Scientific Sessions 2009. Part 2. AB - The American Heart Association (AHA) Scientific Sessions 2009 meeting held in Orlando, FL, USA included topics covering new treatments for cardiovascular disease. This conference report highlights selected presentations on stem cell treatments for ischemia, treatments for hypertension and atherosclerosis, and LDL lowering compounds. PMID- 20024838 TI - AACR-NCI-EORTC--21st International Symposium. Molecular targets and cancer therapeutics--Part 1. AB - The 21st international symposium of the American Association for Cancer Research (AACR), the NCI and the European Organization for Research and Treatment of Cancer (EORTC), held in Boston, included topics covering the development of therapeutics and molecular targets in the field of cancer research. This conference report highlights selected presentations on the development of novel drugs for cancer. Investigational drugs discussed include XL-139 (Bristol-Myers Squibb Co/Exelixis Inc), SPC-3042 (Enzon Pharmaceuticals Inc/ Santaris Pharma A/S), CH-5137291 (Chugai Pharmaceutical Co Ltd), SN-29966/SN-29926/SN-3000 (Proacta Inc), withaferin-A (University of Kansas), Reolysin (Oncolytics Biotech Inc), EZN-3889 and EZN-3892 (Enzon Pharmaceuticals/Santaris Pharma), and CBP-501 (CanBas Co Ltd/Takeda Pharmaceutical Co Ltd). PMID- 20024839 TI - AACR-NCI-EORTC--21st International Symposium. Molecular targets and cancer therapeutics--Part 2. AB - The 21st international symposium of the American Association for Cancer Research (AACR), the NCI and the European Organization for Research and Treatment of Cancer (EORTC), held in Boston, included topics covering the development of therapeutics and molecular targets in the field of cancer research. This conference report highlights selected presentations on the development of novel drugs for cancer. Investigational drugs discussed include RO-506876 (F Hoffmann La Roche Ltd), GDC-0980 (Genentech Inc), EMD-1214063 and EMD-1204831 (Merck Serono SA), AR-mTOR01 and AR-mTOR-26 (Array BioPharma Inc), GSK-2126458 (GlaxoSmithKline plc), EXEL-1415 and EXEL-2008 (Exelixis Inc), FP-1039 (FivePrime Therapeutics Inc), and AV-412 (AVEO Pharmaceuticals Inc). PMID- 20024840 TI - HDAC Inhibitors--CHI's Third Annual Conference. AB - The HDAC Inhibitors meeting, held in Boston, included topics covering new therapeutic developments in the field of HDAC inhibitors. This conference report highlights selected presentations on HDAC inhibitors for the treatment of cancer, inflammation, and neurodegenerative and other CNS diseases. Investigational drugs discussed include several compounds under evaluation by Karus Therapeutics Ltd, OCID-4681 (Orchid Research Laboratories Ltd) and EVP-0334 (EnVivo Pharmaceuticals Inc). PMID- 20024841 TI - Ion Channels as Therapeutic Targets--CHI's Fourth Annual Conference. AB - The Ion Channels as Therapeutic Targets meeting, held in Boston, included topics covering new therapeutic developments in the field of ion-channel drug discovery. This conference report highlights selected presentations on sensory transient receptor potential channels, potassium-channel modulation, as well as methods for chloride-channel screening and channel occupancy evaluation. Human genetic mutations observed in channelopathies and methods to predict the cardiovascular safety of ion-channel modulators were also discussed. PMID- 20024842 TI - RNAi for Developing Targeted Therapeutics--CHI's Third Annual Meeting. AB - The RNAi for Developing Targeted Therapeutics meeting, held in Boston, included topics covering new therapeutic developments in the field of RNAi-based drug discovery. This conference report highlights selected presentations on the reversal of drug resistance using targeted siRNA or small-hairpin RNA and EnGeneIC drug delivery vehicles, the development of siRNA therapeutics for lung diseases, siRNA-based topical microbicides, locked nucleic acid-based oligonucleotides, and company-specific approaches to the development of RNAi therapeutics from Alnylam Pharmaceuticals Inc and MDRNA Inc. In addition to in vivo delivery approaches, novel RNAi-based approaches are also discussed, including dicer substrate siRNA, sd-rxRNA from RXi Pharmaceuticals Corp and a cationic liposome formulation of RNAi. PMID- 20024843 TI - International Drug Discovery Science and Technology--BIT's Seventh Annual Congress. AB - BIT's Seventh Annual International Drug Discovery Science and Technology Congress, held in Shanghai, included topics covering new therapeutic and technological developments in the field of drug discovery. This conference report highlights selected presentations on open-access approaches to R&D, novel and multifactorial targets, and technologies that assist drug discovery. Investigational drugs discussed include the anticancer agents astuprotimut-r (GlaxoSmithKline plc) and AS-1411 (Antisoma plc). PMID- 20024844 TI - Financial Times Global Pharmaceutical & Biotechnology Conference 2009. AB - The Financial Times Global Pharmaceutical & Biotechnology conference, held in London, included topics covering the current and future challenges confronting the pharma and biotech industry, and presented possible solutions to those challenges. This conference report highlights selected presentations on the industry challenges for big pharma companies, diversification as a solution to industry problems, overcoming challenges with collaborations and M&As, and the role of emerging markets in the pharma industry. Other subjects discussed included the expected impact of personalized medicine on the industry, the entry of big pharma into the generics market and the problems that are confronting the small pharma and biotech industry. PMID- 20024845 TI - Current drug patenting for retinal diseases: beyond VEGF inhibitors. AB - An analysis of patent applications that address strategies for the pharmacological treatment of retinal diseases that are not directly related to VEGF inhibition, published under the PCT during the 18-month period from January 2008 to June 2009, is presented. The largest number of therapeutic patent applications focused on attempts to correct visual cycle dysfunctions, complement overactivation or beta-amyloid deposition in drusen to control age-related macular degeneration (AMD). Biomarker-based and genetic diagnostic modalities that assess AMD risk were also frequently claimed in the patent applications, and have become a significant factor in patenting for ocular disorders. The fields of both visual cycle therapy and AMD biomarkers were dominated by non-corporate patent assignees. Diabetic retinopathy has not received as much attention from inventors compared with AMD; retinopathy of prematurity remains a field in which little specific patenting occurs. PMID- 20024846 TI - Palifosfamide, a bifunctional alkylator for the treatment of sarcomas. AB - Ifosfamide is a chemotherapeutic prodrug used in the treatment of several tumor entities, including bone and soft-tissue sarcoma. However, the application of high-dose ifosfamide is not feasible because of severe side effects caused by metabolites. The active metabolite isophosphoramide mustard is not suitable for administration because of chemical instability. ZIOPHARM Oncology Inc, under license from Dekk-Tec Inc, is developing palifosfamide, a formulation of isophosphoramide mustard with tris(hydroxymethyl)aminomethane salt-stabilization (palifosfamide-tris) and previously with lysine-stabilization (palifosfamide lys). Preclinical studies and phase I and I/II clinical trials demonstrated that palifosfamide-tris had an antitumor efficiency comparable or superior to that of ifosfamide. Patients treated with palifosfamide-tris did not display any of the neurotoxic or nephrotoxic side effects associated with ifosfamide. At the time of publication, data from phase II trials were being evaluated and phase III trials were being planned. palifosfamide-tris is expected to be a safer and less toxic alternative to ifosfamide; however, considering other new approaches under investigation for tumors such as sarcoma, such as molecular-based treatment strategies, it is unclear what position palifosfamide-tris might occupy on the market. PMID- 20024847 TI - Left ventricular dysfunction and NT-proBNP levels in patients with one-vessel disease after first ST-elevation myocardial infarction treated with primary coronary angioplasty. AB - BACKGROUND: Cardiovascular diseases continue to be the main cause of death in the general population. Despite the fact that primary percutaneous coronary intervention (PCI) improves prognosis in patients with acute myocardial infarction (AMI), heart failure can still develop in some of these subjects. Measurement of natriuretic peptide levels has been shown to be useful in the detection of left ventricular (LV) dysfunction and the risk stratification of patients with AMI. AIM: To evaluate the role of NT-proBNP in the identification of patients with early systolic LV dysfunction after first AMI with one--vessel disease successfully treated with primary PCI. METHODS: 161 consecutive patients with first ST-elevation AMI treated with primary PCI with stent implantation (occlusion in infarct-related artery was the patient's only lesion) were included. Measurement of NT-proBNP level and echocardiography were performed on the 4-5th day of AMI. RESULTS: In 53 (33%) patients LV dysfunction (defined as EF < 55%) was detected. The remaining 108 (67%) patients had normal LV systolic function. Multivariate regression analysis revealed that NT-proBNP level > 463.4 pg/ml (median level) was the strongest predictor of EF < 55%. A strong negative correlation between EF and NT-proBNP concentration measured on the 4-5th day of AMI was observed (r = -0.54), which increased with decreasing EF value. CONCLUSIONS: Elevated levels of NT-proBNP are the strongest predictor of early LV dysfunction in low-risk patients after first AMI with one-vessel disease treated with primary PCI with complete coronary revascularisation. PMID- 20024848 TI - A new external stent - intimal proliferation and apoptosis in the vein graft in the animal model. AB - BACKGROUND: We have previously showed that the extravascular dacron mesh stent wrapped around a vein graft and implanted into the arterial system prevented the hypertrophy of the graft's wall, impeded the overgrowth of the intima and decreased the proliferation rate of venous graft cellular elements. AIM: To determine the role of cellular proliferation and apoptosis in the process of remodelling in the stent in an animal model in a 12-week period. METHODS: Male sheep (n = 21) received by transplantation the hybrid graft (group 1) or carotid artery radial vein grafts (group 2). A hybrid graft was composed of a radial vein, collagen fibrin glue and highly flexible torlen/dacron mesh tubing. Grafts were retrieved on day 5, 9 and then week 4, 6, 8, 10, 12, respectively. A proliferation process was assessed using a Ki-67 antigen kit. The presence of apoptosis was detected using a TUNEL kit, strictly according to the manufacturer's manual. RESULTS: The number of proliferating cells has presented a decreasing trend in both groups, whereas the mean quantity of apoptotic cells increased over a 12-week period (p < 0.001) in both groups. Proliferation was more prominent during the first 5 weeks in both groups. The trend had a tendency to reverse during the last 7 weeks of observation. The ratio of proliferating to apoptotic cells differed between groups (1.6 vs. 1.9 on day 5 and 0.2 vs. 0.6 in week 12, in group 1 and group 2, respectively). No linear correlation between proliferation and apoptosis was observed (p > 0.05). CONCLUSIONS: Different kinetics in the trico hybrid graft group in comparison with the radial vein graft group was observed, with a more prominent cellular turnover in the trico hybrid graft. Apoptosis in an unprotected vein wall was overcomed by the proliferation process. In trico hybrid vein grafts, beneficial remodelling of the intimal layer was predominantly dependent on inhibition of intimal proliferation rather than the effect of changes of the apoptosis ratio. There was no linear correlation between proliferation and apoptosis in the investigated grafts. PMID- 20024849 TI - Relationship between thoracic fluid content and natriuretic peptide type B in patients with systolic heart failure. AB - BACKGROUND: Thoracic fluid content (TFC) is one of the basic parameters measured by impedance cardiography (ICG). The B-type natriuretic peptide (BNP) is a neuroendocrine mediator produced in the ventricular myocardium and released in response to the increase of wall tension. AIM: To determine the relationship between TFC measured by ICG and BNP serum level in patients with systolic heart failure (HF). METHODS: The study population included 50 patients: a group of 30 patients with systolic HF in functional NYHA class II and III [27 males and 3 females, aged 53 +/- 6 years, with mean left ventricular ejection fraction (LVEF) 23 +/- 6%], and 20 controls without HF symptoms and preserved LVEF. The TFC and BNP serum level were measured on the same day. RESULTS: Mean BNP serum concentration was 521 +/- 882 pg/ml in HF patients and 44 +/- 36 pg/ml in healthy controls (p = 0.02). The TFC values did not differ significantly between the two groups (27.3 +/- 4.5 1/kW in the study group versus 26.3 +/- 2.8 1/kW in control subjects, NS). A significant correlation between TFC and BNP was found in patients with overt HF (r = 0.57, p = 0.001); however, after excluding one patient with exacerbation of HF symptoms, the correlation was non-significant (r = 0.24, p = 0.22). No correlation between these parameters was observed in healthy controls (r = 0.17, p = 0.51). CONCLUSIONS: There was no significant correlation between TFC measured by ICG and BNP serum level in haemodynamically stable patients with HF symptoms. The usefulness of ICG measurements in patients with exacerbated chronic HF needs further investigations. PMID- 20024850 TI - Quality of life and emotional functioning in selected cardiovascular diseases. AB - BACKGROUND: Numerous psychosomatic diseases, especially cardiovascular, are regarded as diseases of modern civilization. Psychological factors play a dominant role in these diseases. Among these factors we can distinguish several different types of emotional functioning and subjective estimation of patients' quality of life (QoL). AIM: To assess the level of QoL and types of emotional functioning among patients suffering from ischaemic heart disease (IHD) and hypertension. METHODS: A group of 160 male patients was subjected to examinations. They were divided into two groups: IHD with hypertension (120 patients) and a control group (40 healthy patients). In the examination the Emotional Control Questionnaire by Brzezinski (KKE) was applied together with Life Quality Test SF-36 and with a personal questionnaire. RESULTS: Approximately 25% of patients had low QoL. The mean QoL was lower than in healthy subjects (p < 0.05). Also parameters of emotional functioning were significantly lower in patients than in controls (p < 0.05). Significant correlations were found between level of QoL and several types of emotional functioning (p < 0.05, p < 0.01 and p < 0.001). CONCLUSIONS: 1. Quality of life level among psychosomatic patients is significantly lower than in healthy individuals. 2. Quality of life level is closely related to emotional functioning of the studied individuals. 3. Among psychosomatic patients lower level of QoL is accompanied by higher intensity of negative emotional functioning schemes. PMID- 20024851 TI - Association between 22 cytokine gene polymorphisms and dilated cardiomyopathy in Macedonian patients. AB - BACKGROUND: Inflammation is an important component in the pathogenesis of many cardiovascular diseases and one of the commonest mechanisms in cardiomyopathy. There have been several studies on the cytokine polymorphism and dilated cardiomyopathy (DCM), but the results obtained were contradictory. AIM: To examine a possible role of 22 cytokine gene polymorphisms in host susceptibility to or protection against DCM in Macedonians. METHODS: In this study 301 healthy unrelated individuals and 52 patients with DCM were studied. Cytokine genotyping was performed by PCR with sequence-specific priming (PCR-SSP) (Heidelberg kit). RESULTS: After the Bonferroni adjustment, the IL-4 -1098/T, IL-4 -1098/T:T, IL 4/TCC, and IL-4/TCC:TTC cytokine genes were positively associated with DCM, while a negative association was identified for IL-4 -1098/G, IL-4 -1098/G:T, IL-1B +3962/C:C, IL-4/GCC, and IL-4/GCC:TTC. CONCLUSIONS: These results suggest that some cytokine gene polymorphisms are significantly associated and affect host susceptibility/resistance to DCM in Macedonians. PMID- 20024852 TI - [Dilation of atypical native coarctation of aorta with stent implantation]. AB - We describe a case of a 21-year-old women with atypical (kinking) native coarctation of aorta. She had arterial hypertension treated pharmacologically. To visualise stenotic segment, standard angiographic planes were not usefull, only right oblique projection showed precisely stenotic segment. Thereafter stent was successfully applied and trans aortic gradient decreased from 30 to 0 mmHg. PMID- 20024853 TI - [Tako-tsubo syndrome without typical anamnesis]. AB - A 77-year-old female in severe emotional distress was admitted to Coronary Care Unit with typical features of acute coronary syndrome, namely severe chest pain, high cardiac necrosis markers and ST-elevation in ECG. Due to the lack of significant changes in coronary arteries in coronary angiography the contractile function of the heart muscle was assessed in echocardiographic examination. That allowed to diagnose tako-tsubo cardiomyopathy. This condition is commonly triggered by stress. This case report puts emphasis on anamnesis, which is often underestimated when diagnosing tako-tsubo syndrome. PMID- 20024854 TI - [Myocarditis in a cachectic female, nonsteroidal anti-inflammatory drugs abuser, in a course of progressive systemic sclerosis]. AB - A case of 70-year-old cachectic female, nonsteroid anti-inflammatory drugs abuser, with progressive systemic sclerosis, who was admitted to our hospital due to joint pain and fatigue is presented. During hospitalisation the patient developed symptoms of acute myocarditis. Angiography of coronary arteries did not reveal narrowing of the vessels. Alimentary supplementation and therapy for heart failure (diuretics, vasodilators, angiotensin-converting enzyme inhibitor and beta-blocker) were used. In repeated echocardiography examinations ejection fraction systematically improved and hemodynamic stabilisation was obtained. Scleroderma, malnutrition, toxicity of nonsteroid anti-inflammatory drugs and infectious agents were considered as a cause of myocarditis. PMID- 20024856 TI - [Atrial fibrillation, other arrhythmias and digestive tract]. PMID- 20024855 TI - [Pulmonary embolism as a first manifestation of synchronous occurrence of two neoplasms]. AB - A case of 67-year-old man with a first episode of acute, unprovoked venous thromboembolism (VTE). Screening for cancer revealed coexistence of two neoplasms: colon sigmoid cancer (operated on 6 weeks after pulmonary embolism onset), and multiple myeloma (treated successfully with thalidomide and dexamethasone). Low molecular weight heparin use as VTE treatment was followed by thromboprophylaxis for myeloma therapy. During a 30-month follow-up period, neither new thromboembolic complications nor cancer recurrence were observed. Overlapping different prothrombotic mechanisms of double malignancy might result in detection of both neoplasms at early stage. PMID- 20024857 TI - [Can ventilatory parameters replace maximal oxygen uptake in the assessment of patients with heart failure]. PMID- 20024858 TI - [Left ventricular non-compaction with papillary muscle involvement - a case report]. AB - Non-compaction myocardium of the left ventricle (LVNC) is a genetically heterogeneous congenital cardiomyopathy characterised by excessive prominent trabeculations and deep intertrabecular recesses which communicate with the left ventricular cavity. Echocardiography plays a pivotal role as a first line diagnostic tool of this rare abnormality. We presented a case of 64-year-old male with LVNC and with papillary muscle involvement. PMID- 20024859 TI - [Acute coronary syndrome in a female patient with multilevel atherosclerosis]. AB - A case of 74-years-old woman who was admitted to Department of Interventional Cardiology due to acute coronary syndrome is presented. The multilevel atherosclerosis was diagnosed. The patient was successfully treated with two stage percutaneous coronary and peripheral revascularisation. During 6-months follow-up patient was in good condition. PMID- 20024860 TI - [Termination of slow ventricular tachycardia which is below ICD detection rate - a case report]. AB - Slow ventricular tachycardia (VT), which is below the detection rate of implantable cardioverter-defibrillator may cause haemodynamical instability, when pharmacological agents or antitachycardia pacing are unsuccessful, electrical cardioversion is necessary. We present another method of termination of slow VT by ICD, in which transcutaneous pacing mimics faster VT and triggers ICD discharge. PMID- 20024861 TI - [Target patients for polyunsaturated omega-3 fatty acids in cardiovascular disease prevention]. PMID- 20024862 TI - Young ambitious cardiologists and scientific research - reasons to get involved. PMID- 20024863 TI - [Use of eptifibatide in patients with acute stent thrombosis, requiring urgent surgical revascularisation - report of 2 cases]. AB - Number of patients requiring intensive antiaggregation therapy in diseases of circulatory system is increasing. Simultaneously many patients are referred to CABG. In the article, based on two case reports, we discuss optimal time frame for performing cardiac surgery, methods to analyze platelet inhibition, and we describe technique of bridging by eptifibatide. PMID- 20024864 TI - Local perforator flaps in soft tissue reconstruction of the upper limb. AB - The quality of reconstruction of soft tissue defects in the upper extremity, resulting either from traumatic injury or tumor excision, has relevant implications both from functional and aesthetic standpoints. Various local and free flaps with more or less consistent donor-site morbidity have been described in the past. The recent introduction of the perforator-based flap concept, has led to an evolution in upper extremity reconstruction, optimizing results at the recipient site whilst minimizing damage to the donor site and, performing this in the simplest way possible. In this study between 2001 and 2008, 31 patients having post-traumatic or post-tumor excision soft tissue defects of the upper limb, were treated using local perforator flaps raised according to two different modalities: "pedicled fasciocutaneous" and "transposition fasciocutaneous/cutaneous". Complete and stable coverage of the soft tissue losses was obtained in all cases with an inconspicuous, only aesthetic, donor site defect. Superficial or partial necrosis of the tip of the flap, due to venous congestion, was observed in 2 cases of "pedicled fasciocutaneous flap". An additional surgical procedure was required in only one of these cases. In our series all 9 patients who had a transposition flap, underwent routinely a preoperative echo color Doppler investigation to identify the main perforators. In only one case did the Doppler investigation fail to accurately locate the perforator. Local perforator flaps allow the coverage of medium size defects in the upper extremity, can be raised with a relatively simple surgical technique, have a high success rate and good aesthetic results without functional impairment. In the light of this they can be considered among the surgical choices to resurface complex soft tissue defects of the upper extremity. Preoperative identification of the perforators in case of "transposition flaps" greatly facilitates the operation. In our experience echo color Doppler investigations provided reliable results. PMID- 20024865 TI - Commentary on the article of C. Radtke et al.: efficient production of transfected human keratinocytes under serum free and feeder layer free conditions. PMID- 20024867 TI - Skin rejuvenation. PMID- 20024868 TI - Overview of facial aging. AB - Facial aging is a multidimensional, multifactorial process. The aging face has traditionally been treated by each specialty in a different manner. However, by understanding the process from the perspective of different specialties, each physician may better treat the spectrum of facial aging. Whether or not the facial plastic surgeon injects products to restore volume, uses lasers to resurface the epidermis and dermis, incorporates cosmeceuticals to enhance and maintain improvements in the skin integrity and appearance, or relaxes muscles with botulinum toxins, he or she can best advise patients and address facial aging by having a functional understanding of these various modalities. With this knowledge, the facial plastic surgeon can parse the component of facial aging that enables him or her to correct each with the appropriate treatment. PMID- 20024869 TI - Cosmeceuticals for rejuvenation. AB - Facial plastic surgeons are frequently called upon to enhance the appearance of their patients. Typically, this involves surgery of the face or neck. Increasingly, this may also require the use of injectables such as botulinum toxins, fillers, and volumizers. Lasers and light sources are used to enhance the appearance of the surface of the skin. Each of these modalities will help to improve the patient but are limited by the fact that they require the patient to be treated in the office. With the advent of effective topical agents, patients are increasingly able to improve their appearance with the use of prescription and nonprescription topical products. It is incumbent upon the facial plastic surgeon to have an understanding of these topical agents so that they can assist the patient in deciding what does and does not make sense. In addition, many cosmetic physicians will dispense these products in their office, and it behooves them to understand the various products available and what will likely be produced in the future. PMID- 20024870 TI - Intense pulsed light: what works, what's new, what's next. AB - Intense pulsed light technology has revolutionized the noninvasive treatment of a whole variety of cutaneous cosmetic problems. Today's machines are safer, more diverse, less painful, and readily available. Intense pulsed lights can be used to treat vascular and pigmented lesions and are used for facial rejuvenation and the removal of unwanted hair. Newer, less powerful home-device intense pulsed light sources will not replace those used in a physician's office but will augment the use of today's more powerful medical office systems. PMID- 20024871 TI - Dermabrasion and microdermabrasion. AB - Dermabrasion and microdermabrasion are facial resurfacing techniques that mechanically ablate aged or damaged skin to promote reepithelialization. Although the act of physically abrading the skin is common to both procedures, dermabrasion and microdermabrasion employ different instruments and are distinct in their technical executions. Dermabrasion completely removes the epidermis and penetrates to the level of the papillary or reticular dermis, inducing remodeling of the skin's structural proteins. Microdermabrasion only removes the uppermost layer of the epidermis, accelerating the natural process of exfoliation. Both techniques can result in clinically significant improvements in the appearance of skin and may become increasingly relevant among practitioners as new applications are developed. PMID- 20024872 TI - Treatment of facial scarring: lasers, filler, and nonoperative techniques. AB - Many techniques have been described to improve facial scars. In this review we address nonoperative strategies to manage facial scarring. Scar characteristics and anatomic location are discussed. The following nonoperative techniques are comprehensively examined: pulsed dye laser (PDL), subcision, fat transplantation, collagen injection, dermabrasion, steroid injection, and fillers. The PDL is advocated to flatten and decrease the volume of hypertrophic scars while improving texture and pliability. Dermabrasion is used to blend acne scars into the surrounding facial skin by subtly improving their contour. Fat transplantation, collagen, and filler injection is recommended in the treatment of acne depressed scars and traumatic scars with resulting contour deformity. PMID- 20024873 TI - Treatment of laser complications. AB - Modern lasers and light-based sources that were developed based on the theory of selective photothermolysis are capable of destroying specific tissue targets while minimizing the risk of scarring and pigmentary changes. This is accomplished through the use of a wavelength and pulse duration that is best absorbed by a specific chromophore such as melanin or hemoglobin. However, not all lasers and light sources adhere to this principle. Continuous wave (CW) lasers are least selective and may produce unwanted tissue damage and scarring through heat conduction to normal skin. Quasi-CW lasers limit excessive thermal destruction by delivery of a series of brief laser pulses but still pose a higher risk of nonspecific tissue damage and thermal injury. The pulsed and Q-switched (QS) systems adhere most closely to the principles of selective photothermolysis and result in the highest degree of selective destruction with the lowest risk of scarring from excessive thermal diffusion. Certainly, any laser system potentially can result in scarring and tissue damage when used incorrectly; therefore, adequate operator education and skill are essential. Side effects and complications that occur as a consequence of laser treatment can be significantly reduced if diagnosed and treated in an expeditious manner. PMID- 20024874 TI - Complications of hyaluronic acid fillers. AB - Hyaluronic acid filler use, user groups, and indications have expanded significantly in the past several years. This group of fillers is extremely safe in experienced hands. Complications are infrequent but can be devastating. There can be no substitution for recognized and specific training. Prompt recognition and proper treatment of serious complication can moderate and even prevent serious sequelae. This article describes the most frequent and serious complications, their prevention, and treatment. PMID- 20024875 TI - Chemical peels: what's new and what isn't new but still works well. AB - Chemical peeling is becoming increasingly popular for the treatment of skin rejuvenation where it can improve damaged skin and fine wrinkles. The basic procedure aims at obtaining a controlled chemical burn of the epidermis and/or dermis. This results in epidermal regeneration and postinflammatory collagen neoformation with remodeling of collagen and elastic fibers and deposition of glycosaminoglycans in the dermis. Various chemicals have been used as peeling agents, of which the most used are the alpha-hydroxy acids, such as glycolic acid, or beta-hydroxy acids, such as salicylic acid. The choice of the compound is linked to the different indications and to the depth of the desired peeling. Phenol is still the best agent for deep peeling but requires specific indications, prescription, and post-peeling care. Combination of different compounds is one innovation in the field of chemical peelings. Further controlled studies are necessary to set up specific guidelines. PMID- 20024876 TI - Current and new treatments of photodamaged skin. AB - Patients with photodamaged skin need guidance in selecting treatment plans that optimize outcomes, minimize downtime, and reduce adverse effects. The gold standard among cosmeceuticals is the topical retinoids, such as tretinoin. A topical formulation of folic acid and creatine appears to be a viable treatment option for the treatment of photodamaged skin. The use of specific topical cosmeceuticals in combination with nonablative photorejuvenation is recommended in choosing modalities that address the concerns of the patient. A combination of intense pulsed light (IPL), low-intensity diode light, and biostimulating drugs has been shown to provide results superior to those of IPL alone for photorejuvenation. Photodynamic therapy (PDT) with 5-aminolevulinic acid (5-ALA) is the treatment of choice for type C photodamage. Low-strength 5-ALA (1 to 2%) applied several times, every 10 to 15 minutes, and incubated for 30 to 60 minutes with 550 to 630 nm, 530 to 1200 nm, or 570 to 1200 nm light activation improved hyperpigmented lesions, skin smoothing, and skin elasticity with high patient satisfaction. The use of 0.5% liposome-encapsulated 5-ALA spraying has been shown to be an alternative to 20% 5-ALA in a cream base in patients undergoing photorejuvenation. Adipose-derived stem cells and their derived secretory factors may have potential as treatments of photodamage. PMID- 20024877 TI - [Transfer of patients between a unit of geriatric psychiatry and general hospitals: a one-year analysis]. AB - BACKGROUND AND OBJECTIVE: Patients with psychogeriatric disorders normally are affected by comorbid medical illnesses. In this context the question where to set therapeutic priorities often arises. With regard to that issue frequency of and indications for patient transfer from general hospital to a psychogeriatric unit and vice versa were evaluated in this study. PATIENTS AND METHODS: Retrospective analysis of all admissions during one year in a geriatric psychiatry department of a regional hospital for psychiatry and neurology. RESULTS: 1005 episodes of treatment were analysed: 65.9 % female patients, mean age 76.7 + or - 9.3 years, mean length of stay 37.2 + or - 25.1 days; main diagnoses: organic mental disorders 50.9 %, affective disorders 30.8 %, others 18.3 %. In 330 cases patients were transferred, either from general hospital to the psychogeriatric unit (n = 164) or vice versa (n = 100), or transfer took place back and forth (n = 66). In the overall 166 transfers to general hospitals acute general medical conditions were the most frequent causes (62.7 %), with cardiovascular and respiratory diseases representing the most common diagnoses (24.1 and 13.3 % of all transfers, respectively). CONCLUSION: Although the results presented here originate from one single region and thus cannot be taken as representative for Germany as a whole they indicate a considerable overlap of the clientele of general hospitals and psychogeriatric units. In view of expected demographic changes which presumably will result in a growing proportion of persons with (psycho)geriatric disorders, cooperation between general hospitals, psychogeriatric departments and outpatient treatment has to be intensified with high priority. PMID- 20024878 TI - [Neuroendocrine tumor of the esophagus - successful endoscopic treatment of a very rare entity]. AB - HISTORY AND CLINICAL FINDINGS: A 77-year old women presented with dysphagia and loss of weight for two months INVESTIGATIONS, TREATMENT AND DIAGNOSIS: Endoscopy revealed a polypoid tumor was found in the upper third of the esophagus. It was resected endoscopically after a deeper infiltration of the esophageal wall and enlarged lymphadenopathy or distant metastasis had been ruled out. Histology showed a highly differentiated neuroendocrine tumor of the esophagus with beginning infiltration of the submucosa. It was completely resected. COURSE: The follow-up of 48 months showed no local recurrence or distant metastasis. This case report is the second description of a successful endoscopic treatment of a neuroendocrine tumor of the esophagus. CONCLUSION: EMR is a diagnostic and therapeutic tool in case of mucosal lesions of the gastrointestinal tract. Highly differentiated localized NET of the esophagus are very rare. PMID- 20024879 TI - [Metabolic disturbances in liver cirrhosis (part 2), hepatogenous diabetes: diagnostic aspects and treatment]. AB - Uip to 20% of patients with liver cirrhosis develop hepatogenous diabetesdue to the hepatocellular functional loss and insulin resistance. Optimizing diabetic metabolic conditions is not only important to avoid typical late complications of diabetes, but also cirrhosis-associated complications e.g. gastrointestinal bleeding, hepatic encephalopathy or the occurence of hepatocellular carcinoma. So far there have beenno recommendations orguidelines for the diagnosis and treatmentof hepatogenous diabetes. The medical teatment of a diabetic condition is mainly influenced by its side effects. The risk of hypoglycemia must be considered carefully during drug treatment, especially in patients with chronic alcohol abuse. Suitable oral antidiabetics are glinides and short-acting sulfonylureas or possibly meal-related insulin administration with short-acting insulins or rapid-acting insulin analogues. Biguanide and PPAR-gamma agonists are contraindicated because of side effects in liver cirrhosis. Regarding basic treatment, an adequate daily energy and protein supply should be ensuredbecause the majority of patients with liver cirrhosis are malnourished. PMID- 20024880 TI - [Mediquiz case 2943. 52-year-old woman after removal of an inguinal central venous line]. PMID- 20024881 TI - [Lipoid pneumonia - an underestimated syndrome]. AB - Lipoid pneumonia, first described by Laughlen 1925 may be classified as endogenous or exogenous. The endogenous form is seen when fat is deposited into the lung tissue. It is usually associated with proximal obstructive lesions, necrotic tissue after radio- or chemotherapy, with lipid storage disease or hyperlipidemia . Exogenous lipoid pneumonia results from inhaling or aspirating animal, vegetable or mineral oil. There are usually some underlying neurological defects or esophageal abnormalities. Patients may present with cough, sputum, hemoptysis and chest pain or may be asymptomatic. There is no classic chest film appearance: it may appear as diffuse airspace infiltration or localized consolidation simulating tumour. Computed tomography is diagnostically helpful and shows hypodense areas measuring from -100 to - 30 Hounsfield units. Bronchoscopic biopsies are mandatory for histological confirmation of the diagnosis. Treatment of exogenous lipoid pneumonia has always been conservative by discontinuing the use of oil, correction of underlying defects that may favor aspiration and treatment of intercurrent pneumonia. Other measures, for example corticosteroid therapy, are of uncertain benefit. Complications of lipoid pneumonia that worsen prognosis are recurrent bacterial pneumonias including nontuberculous mycobacteria or aspergillus, or lung cancer that has developed in areas of pre-existing exogenous lipoid pneumonia. PMID- 20024882 TI - [Entecavir - options and obstacles of an effective treatment for chronic hepatitis B]. AB - The level of HBV-DNA is a crucial determinant of the progression to liver cirrhosis or hepatocellular carcinoma. Thus the effective suppression of HBV-DNA below the limit of detection of high sensitive assays is a major aim of treatment. This aim will be achieved in almost all patients with current available direct antiviral drugs. However, the development of drug resistance remains a main challenge for the future. Entecavir has a good profile regarding both antiviral efficacy and resistance profile in the treatment of previously untreated patients. After five year on Entecavir more than 90 % of HBe-Ag positive patients had achieved HBV-DNA below 300 copies/ml and resistance developed in only about 1 %. However, patients in whom there had been previous resistance to lamivudine have lower treatment responses and higher resistance rates because only one or two additional mutations in the HBV-polymerase gene are required for the development of entecavir resistance in contrast to three mutations required in treatment naive patients. In summary, Entecavir adds a new milestone in the treatment of chronic hepatitis B. PMID- 20024883 TI - [Liability of a hospital and its medical director for errors committed by one of their self-employed consulting doctors]. PMID- 20024884 TI - [Clinical course of severe influenza-A/H1N1 infection with fulminant lung failure]. PMID- 20024885 TI - Effects of physical efforts on injury in elite soccer. AB - In this study, the influence of physical efforts on occurrence of match injury in a professional soccer club was investigated. Computerised motion-analysis was used to measure the physical efforts of players during 10 injury situations. Total distance and those covered at different movement intensities were measured across the 5-min period preceding injury. If the final run preceding injury involved a high-intensity action (HIA), the distance, duration and speed of the effort and the recovery time between this and the penultimate HIA were measured. To determine the influence of these physical efforts, the results were compared to a normative profile for players computed from data across 5 games for the same variables; habitual distances covered over a 5-min period and characteristics of and recovery time between HIA. Compared to the normative profile, no differences were reported in physical characteristics during the period leading up to injury or for HIA although the latter were substantially higher in intensity (duration and distance). A lower than normal recovery time between HIA prior to injury was observed (35.6+/-16.8 s vs. 98.8+/-17.5 s, p=0.003). Within the limitations of the small sample, these findings may aid in further understanding injury and physical performance in elite soccer. PMID- 20024886 TI - Implementation of a critical incident reporting system in a neurosurgical department. AB - BACKGROUND: Critical incident monitoring is an important tool for quality improvement and the maintenance of high safety standards. It was developed for aviation safety and is now widely accepted as a useful tool to reduce medical care-related morbidity and mortality. Despite this widespread acceptance, the literature has no reports on any neurosurgical applications of critical incident monitoring. We describe the introduction of a mono-institutional critical incident reporting system in a neurosurgical department. Furthermore, we have developed a formula to assess possible counterstrategies. METHODS: All staff members of a neurosurgical department were advised to report critical incidents. The anonymous reporting form contained a box for the description of the incident, several multiple-choice questions on specific risk factors, place and reason for occurrence of the incident, severity of the consequences and suggested counterstrategies. The incident data was entered into an online documentation system (ADKA DokuPik) and evaluated by an external specialist. For data analysis we applied a modified assessment scheme initially designed for flight safety. RESULTS: Data collection was started in September 2008. The average number of reported incidents was 18 per month (currently 216 in total). Most incidents occurred on the neurosurgical ward (64%). Human error was involved in 86% of the reported incidents. The largest group of incidents consisted of medication related problems. Accordingly, counterstrategies were developed, resulting in a decrease in the relative number of reported medication-related incidents from 42% (March 09) to 30% (September 09). CONCLUSIONS: Implementation of the critical incident reporting system presented no technical problems. The reporting rate was high compared to that reported in the current literature. The formulation, evaluation and introduction of specific counterstrategies to guard against selected groups of incidents may improve patient safety in neurosurgical departments. PMID- 20024887 TI - Long-term follow-up of a non-infantile desmoplastic ganglioglioma. AB - Desmoplastic gangliogliomas are mixed cerebral tumors traditionally reported in infants. However, a few non-infantile cases have been documented. A case of a desmoplastic ganglioglioma in a 16-year male is presented. The patient reported severe headaches. Radiological examination revealed a large mass occupying the right frontal lobe. The lesion was totally excised. Histopathological examination confirmed the diagnosis of a desmoplastic ganglioglioma. The postoperative course was excellent. At the 10(1/2) year follow-up there was no evidence of tumor recurrence. Although desmoplastic gangliogliomas have aggressive features, complete surgical removal is the treatment of choice obviating the need for adjuvant therapy. PMID- 20024888 TI - Current techniques in preoperative imaging for abdomen-based perforator flap microsurgical breast reconstruction. AB - Perforator-based microsurgical reconstruction of the breast has steadily increased since the introduction of the technique in the 1990s. The procedure appears to offer less postoperative pain, lower abdominal morbidity, and better preservation of the rectus muscles than the more conventional flaps. However, the major disadvantage of these flaps that they can be difficult to harvest, resulting in a longer operative times. The challenges in flap dissection are a result of the variability in the vascular anatomy of the deep inferior epigastric artery (DIEA) and its perforating branches through the rectus muscle. The location, number, and caliber of the perforators and the intramuscular trajectory of the DIEA branches vary greatly not only from individual to individual, but from one hemiabdomen to the other. The establishment of a presurgical map of the vessels on the abdomen facilitates surgical planning and may decrease operating room time, reduce intraoperative complications, and possibly improve outcomes. This article reviews the available techniques for preoperative planning with the currently available imaging modalities of handheld Doppler, color Doppler (duplex) ultrasound, computed tomography angiography, and magnetic resonance angiography. PMID- 20024889 TI - 13C-NMR detection of STD spectra. AB - We have investigated the use of (13)C for the detection of saturation transfer difference (STD) NMR spectra. By detecting the STD spectrum in the (13)C channel it is possible to eliminate the residual water signal in the STD-NMR spectrum. We have employed an INEPT transfer in order to shift the magnetization from the proton channel to (13)C. As a sample system to check our method we have used human serum albumin and phenylalanine. We have shown that such a transfer can be accomplished and gives reasonable signal intensities. PMID- 20024890 TI - Tear analysis of ascorbic acid, uric acid and malondialdehyde with capillary electrophoresis. AB - Tears have a significant role in antioxidant defense in ocular tissues and since their collection is quick and noninvasive, their analysis would facilitate monitoring of pathophysiological changes. However, their low volume and low content of antioxidants makes analysis difficult; methods of high sensitivity are needed. In this paper, we present a method for tear analysis of two antioxidant molecules (ascorbic and uric acid) and of a lipid peroxidation indicator (malondialdehyde) with capillary electrophoresis. Tears were collected with Schirmer strips, extracted with a low-pH phosphate buffer, centrifuged through membrane filters and an antioxidant was added. They were stable at -70 degrees C for 15 days. After pilot experiments, optimum electrophoretic separation was achieved in a 25 mM borate buffer, pH 10.0, containing 100 mM sodium dodecyl sulfate at 25 degrees C and 20 kV. The developed method has good repeatability (<5% RSD), precision (<15% relative error values) and high sensitivity (LLOQ values of 20, 2.3 and 2.5 microM for ascorbate, urate and malondialdehyde, respectively). It was applied to the analysis of tears from healthy individuals and the antioxidant levels are in agreement with those obtained with other techniques. This method might serve as a tool to clarify the role of endogenous antioxidants in the pathophysiology of ocular diseases. PMID- 20024891 TI - An LC/MS method for the quantitative determination of 7alpha-OH DHEA and 7beta-OH DHEA: an application for the study of the metabolism of DHEA in rat brain. AB - Dehydroepiandrosterone (DHEA) is an important neurosteroid with neuronal protection and memory enhancement functions. 7alpha-OH DHEA and 7beta-OH DHEA are the two important metabolites of DHEA in the brain. We have developed an LC/MS method to quantitatively analyze 7alpha-OH DHEA and 7beta-OH DHEA. Chromatographic separation was carried out on a C18 column with gradient elution using mobile phases of formic acid in acetonitrile and in water formic acid. Mass spectral detection was performed with a ThermoFinnigan LCQ advantage quadruple ion trap mass spectrometer with electrospray ionization. Positive ion chromatograms were acquired using single ion monitoring. The protonated molecule was 305 m/z, but the most abundant ion (269 m/z) was used for quantification. This method was validated and applied to investigate the 7-hydroxylation of DHEA. When incubating DHEA with rat brain microsomes, both 7alpha-OH DHEA and 7beta-OH DHEA were observed, but 7alpha-OH DHEA was the major metabolite. PMID- 20024892 TI - Assessment of COMT isolation by HIC using a dual salt system and low temperature. AB - Sodium citrate (SC) and low temperatures between 7 and 5 degrees C are effective in suppressing aggregation of proteins and may be beneficial to be included during a purification process. In this work, we analyzed the application of dual salt system, ammonium sulfate (AS) and SC on binding and elution conditions of recombinant hSCOMT on typical HIC sorbents. Specifically in butyl and octyl supports, the use of, respectively, 300 mM AS/200 mM SC and 25 mM AS/25 mM SC in the loading buffer resulted in complete binding of COMT. Elution was obtained by decreasing the ionic strength to 0 M of salt. For the delineate goal, it also favorably increased the support chain length while a consequent decrease in the dual ionic strength was observed for hSCOMT retention. In the presence of dual salt systems octyl media exhibited classic HIC behavior, good protein selectivity, an excellent purification factor and reduced denaturation effects of hSCOMT observed with higher salt concentrations. Also the inclusion of temperature control during the elution step appears to be advantageous for greater activity recovery without enzyme aggregation. In fact, these results could allow the prediction of most stabilizing conditions for this termolabile enzyme on the chromatographic stage, regarding salt types and therefore effectiveness to improve HIC selectivity and desirable purity on the target fractions. PMID- 20024893 TI - Reliability of knee joint range of motion and circumference measurements after total knee arthroplasty: does tester experience matter? AB - BACKGROUND AND PURPOSE: Two of the most utilized outcome measures to assess knee joint range of motion (ROM) and intra-articular effusion are goniometry and circumference, respectively. Neither goniometry nor circumference of the knee joint have been examined for both intra-tester and inter-tester in patients with total knee arthroplasty (TKA). The purpose of this study was to determine the intra-tester and inter-tester reliability of active and passive knee joint ROM and circumference in patients with TKA when administered by physiotherapists (testers) with different clinical experience. METHOD: The design was an intra tester, inter-tester and intra-day reliability study. Nineteen outpatients (10 females) having received a TKA were examined by an inexperienced and an experienced physiotherapist. Following a standardized protocol, active and passive knee joint ROM and circumference measurements were obtained using a universal goniometer and a tape measure, respectively. To establish reliability, intraclass correlation coefficients (ICC(2,1)) and smallest real difference (SRD) were calculated. RESULTS: The knee joint ROM and circumference measurements were generally reliable (ICC > 0.8) within and between physiotherapists (except passive knee extension). Changes in knee joint ROM of more than 6.6 degrees and 10 degrees (except active knee flexion) and knee joint circumference of more than 1.0 cm and 1.63 cm represent a real clinical improvement (SRD) or deterioration for a single individual within and between physiotherapists, respectively. Generally, the experienced tester recorded larger knee joint ROM and lower circumference values than that of the inexperienced tester. CONCLUSIONS: In clinical practice, we suggest that repeated knee goniometric and circumferential measurements should be recorded by the same physiotherapist in individual patients with TKA. Tester experience appears not to influence the degree of reliability. PMID- 20024895 TI - The structure of common mental disorders: a replication study in a community sample of adolescents and young adults. AB - Previous research suggests that patterns of comorbidity of common mental disorders among adults are best reflected by a hierarchical three-factor structure with two correlated factors ('anxious-misery' and 'fear') summarized in a second-order 'internalizing' factor and one 'externalizing' factor. This three factor structure has not been examined yet in a sample of adolescents and young adults.A representative sample of 3021 adolescents and young adults (baseline age 14-24) were prospectively followed over 10 years. Mental disorders were assessed according to the Diagnostic and Statistical Manual of Mental Disorders, Fourth Edition (DSM-IV) by using the standardized Munich Composite International Diagnostic Interview. Ten mental disorders (major depressive episode, dysthymia, generalized anxiety disorder, social phobia, specific phobia, agoraphobia, panic disorder, alcohol dependence, drug dependence, antisocial personality) were fitted to a series of Confirmatory Factor Analysis models using: (1) 12-month data, and (2) lifetime data from a person-year data set.The three-factor model showed good fit to the observed data in our sample both when 12-month diagnoses and lifetime-to-date diagnoses from a person-year data file were used; yet the higher-order 'internalizing' factor summarizing 'anxious misery' and 'fear' had to be omitted.The three-factor model could be replicated in a sample of adolescents and young adults with the exception that the second-order 'internalizing' factor was not consistent with the data. Further research is necessary to provide more complete insight into the structure of mental disorders by examining the stability of the structure of mental disorders in different developmental stages (ages) and by using a more extensive set of mental disorders. PMID- 20024894 TI - Comparison of three different extraction methods and HPLC determination of the anthraquinones aloe-emodine, emodine, rheine, chrysophanol and physcione in the bark of Rhamnus alpinus L. (Rhamnaceae). AB - INTRODUCTION: Rhamnus alpinus L. (Rhamnaceae), a traditional plants in the flora of the Abruzzo region, is known to contain active anthraquinone secondary metabolites. However, the content of anthraquinones varies among R. alpinus samples depending on collection season and site. Thus, using simple, reliable and accurate analytical methods for the determination of anthraquinones in R. alpinus extracts allows comparative study of different methods of extraction. OBJECTIVE: After a partial validation of an HPLC method for the simultaneous determination of five anthraquinones, aloe-emodine, rheine, emodine, chrysophanol and physcione, in the bark of R. alpinus, we compared three different methods of extraction. METHODOLOGY: Anthraquinones were extracted from the bark of R. alpinus using different techniques (methanol maceration, ultrasonic and supercritical CO(2) extraction). Separation and quantification of anthraquinones were accomplished using a reversed-phase C(18) column with the mobile phase of H(2)O-methanol (40 : 60, v/v, 1% formic acid) at a wavelength of 254 nm. The qualitative analyses were also achieved at wavelength of 435 nm. RESULTS: All calibration curves were linear over the concentration range tested (10-200 mM) with the determination coefficients >or=0.991. The detection limits (S/N = 3) were 5 mM for each analytes. All five anthraquinones were found in the samples tested at concentrations reported in experimental data. CONCLUSION: The described HPLC method and optimised extraction procedure are simple, accurate and selective for separation and quantification of anthraquinones in the bark of R. alpinus and allow evaluation of the best extraction procedure between the tested assays. PMID- 20024896 TI - In vitro studies on the influence of surface modification of Ni-Ti alloy on human bone cells. AB - The in vitro cell behavior on Nitinol after different surface treatments was investigated. As references samples, commercially pure titanium (cpTi) and bioactive titanium were used. The surface treatments influenced the topography, surface energy, crystallographic structure, ion release, chemistry, and ability to form apatite layer from simulated body fluids. Regardless of the surface treatment, the bioactivity study showed that the kinetics of apatite film formation was similar for all tested samples. No clear indication of the surface characteristics influence on the ability for calcium-phosphate precipitation was evident. Cell activity studies showed that ground nickel titanium, spark oxidized and thermally oxidized (at 400 degrees C and below) had higher cellular activity and caused increased alkaline phosphatase (ALP) and osteocalcin (OC) expression which was comparable to control tissue culture plastic and titanium reference samples. Regardless of surface modifications, preimmersion of the samples in media for 72 h resulted in cell proliferation at the same level for all samples. Therefore, it can be concluded that preconditioning of samples alters surface properties and modulates the cell response regardless of the initial surface treatment and its properties. Moreover, a detrimental effect on cell response was observed after 7 and 14 days in culture for alkali treated samples. This was attributed to a high surface nickel concentration and a high nickel ion release rate from these surfaces. PMID- 20024897 TI - Platycodin D inhibits adipogenesis of 3T3-L1 cells by modulating Kruppel-like factor 2 and peroxisome proliferator-activated receptor gamma. AB - In this study, platycodin D was found to inhibit intracellular triglyceride accumulation in 3T3-L1 cells with an IC(50) of 7.1 microM. The expression levels of genes involved in lipid metabolism such as fatty-acid-binding protein 4 and lipoprotein lipase were significantly downregulated following treatment with platycodin D. Treatment with platycodin D also resulted in a reduction of Peroxisome proliferator-activated receptor(PPAR)gamma expression and its binding to target DNA sequence. Among the various upstream regulators of PPARgamma, the expression of Kruppel-like factor(KLF)2, an anti-adipogenic factor, was significantly upregulated following platycodin D treatment. When the upregulation of KLF2 was inhibited by KLF2 siRNA, the expression and binding of PPARgamma to its target sequence were significantly recovered under these conditions. The results of this study suggested that anti-adipogenic effect of platycodin D involves the upregulation of KLF2 and subsequent downregulation of PPARgamma. PMID- 20024900 TI - Are postoperative hearing results better with titanium ossicular reconstruction prostheses? PMID- 20024902 TI - Serum anti-glycan antibodies predict complicated Crohn's disease behavior: a cohort study. AB - BACKGROUND: A high proportion of patients with Crohn's disease (CD) over time develop complications like fistulae and strictures, requiring surgery. We tested a panel of antiglycan antibodies for predicting the occurrence of complications and CD-related surgery in an adult patient cohort. METHODS: Serum samples of 149 CD patients of the German inflammatory bowel disease (IBD) network were tested for the presence of anti-laminarin IgA (Anti-L), anti-chitin IgA (Anti-C), anti chitobioside IgA (ACCA), anti-laminaribioside IgG (ALCA), anti-mannobioside IgG (AMCA), and anti-Saccaromyces cerevisiae IgG (gASCA) carbohydrate antibodies by enzyme-linked immunosorbent assay (ELISA) (IBDX(R) panel, Glycominds, Lod, Israel) in a blinded fashion. Clinical data were available on occurrence of complicated disease or CD-related surgery as well as disease activity, onset, and location. RESULTS: The median follow-up of the patients without any previous complication or surgery at time of sample procurement was 53.7 months. Overall, 26.3% developed a complication and 17.1% underwent CD-related surgery, respectively. Positivity for gASCA, AMCA, ACCA, and Anti-L alone or an increasing frequency of positive serum antibodies independently predicted a faster progression toward a more severe disease course. Once a complication or surgery had occurred only positivity for Anti-L or more than 3 markers out of the whole panel indicated progression to an additional surgery or complication. The antibody status of most patients remained stable over time. CONCLUSIONS: This is the first study showing the clinical value of serum antiglycan antibodies for prediction of a more complicated disease course in adult patients with CD. PMID- 20024903 TI - Management of inflammatory bowel diseases: a changing paradigm. Can we finally change the disease course? PMID- 20024904 TI - Variants at the 3p21 locus influence susceptibility and phenotype both in adults and early-onset patients with inflammatory bowel disease. AB - BACKGROUND: To date, a number of high-profile studies have yielded over 50 inflammatory bowel disease (IBD) disease genes/loci. The polymorphisms rs9858542 (BSN) and rs3197999 (MST1), on 3p21 locus, have been found associated with susceptibility to IBD. We aimed to replicate these associations in adult and early-onset cohorts of IBD Italian patients, by analyzing also potential gene gene interactions with variants in NOD2/CARD15, IL23R, ATG16L1, and IRGM genes, and investigating genotype-phenotype correlation. METHODS: In all, 1808 patients with IBD, 855 with Crohn's disease (CD) and 953 with ulcerative colitis (UC), including 539 patients with their initial diagnosis <19 years of age, and 651 controls were analyzed for SNPs rs9858542 and rs3197999. RESULTS: BSN and MST1 were significantly associated with either CD (P(rs9858542) 2.5 x 10(-7); P(rs3197999) 3.9 x 10(-7)), and UC (P(rs9858542) = 3.1 x 10(-4); P(rs3197999) = 8 x 10(-4)). Prevalence of these variants was significantly increased in both adult and early-onset IBD patients. After stepwise logistic regression, the 2 variants were associated in adult UC with distal colitis (P(rs9858542) = 0.013, odds ratio [OR] = 2.04, 95% confidence interval [CI] = 1.16-3.59; P(rs3197999) = 0.018, OR 1.9, 95% CI 1.2-3.3), while the rs3197999 variant was inversely associated with occurrence of extraintestinal manifestations in adult CD(P = 0.017, OR 0.6, 95% CI 0.4-0.9). CONCLUSIONS: We confirmed the association of BSN and MST1 with IBD susceptibility, either in the adult or the early-onset cohorts. These variants appeared to influence either the distal location of the disease in the UC cohort and extraintestinal manifestations in CD patients. PMID- 20024905 TI - Enhanced translocation of bacteria across metabolically stressed epithelia is reduced by butyrate. AB - BACKGROUND: The gut microflora in some patients with Crohn's disease can be reduced in numbers of butyrate-producing bacteria and this could result in metabolic stress in the colonocytes. Thus, we hypothesized that the short-chain fatty acid, butyrate, is important in the maintenance and regulation of the barrier function of the colonic epithelium. METHODS: Confluent monolayers of the human colon-derived T84 or HT-29 epithelial cell lines were exposed to dinitrophenol (DNP (0.1 mM), uncouples oxidative phosphorylation) + Escherichia coli (strain HB101, 10(6) cfu) +/- butyrate (3-50 mM). Transepithelial resistance (TER), and bacterial internalization and translocation were assessed over a 24 hour period. Epithelial ultrastructure was assessed by transmission electron microscopy. RESULTS: Epithelia under metabolic stress display decreased TER and increased numbers of pseudopodia that is consistent with increased internalization and translocation of the E. coli. Butyrate (but not acetate) significantly reduced the bacterial translocation across DNP-treated epithelia but did not ameliorate the drop in TER in the DNP+E. coli exposed monolayers. Inhibition of bacterial transcytosis across metabolically stressed epithelia was associated with reduced I-kappaB phosphorylation and hence NF-kappaB activation. CONCLUSIONS: Reduced butyrate-producing bacteria could result in increased epithelial permeability particularly in the context of concomitant exposure to another stimulus that reduces mitochondria function. We speculate that prebiotics, the substrate for butyrate synthesis, is a valuable prophylaxis in the regulation of epithelial permeability and could be of benefit in preventing relapses in IBD. PMID- 20024908 TI - Cytomics for discovering drugs. PMID- 20024907 TI - Phenotypes of stem cells from diverse origin. AB - Stem cells have turned into promising tools for studying the mechanisms of development, regeneration, and for cell therapy of various disorders. Stem cells are found in the embryo and in most adult tissues participating in endogenous tissue regeneration. They are capable of autorenovation, often maintain their multipotency of differentiation into various tissues of their germ line and are, therefore, ideal candidates for cellular therapy taken that they can be unequivocally identified and isolated. In this review, we report stem cell marker expression used for identification of various stem cell lineages, including very small embryonic stem cells, neural, hematopoietic, mesenchymal, epithelial and limbal epithelial stem cells, endothelial progenitor cells, supra-adventitial adipose stromal cells, adipose pericytes, and cancer stem cells. These cells usually cannot be distinguished by a single stem cell marker, because their expression partially overlaps between lineages. Recent advances in flow cytometry allowing the simultaneous detection of various markers have facilitated stem cell identification for clinical diagnosis and research. So far experimental evidence suggests the existence of cells with different properties, i.e., the capability to different in various cell types. Several studies indicate that expression of classical markers for stem cell classification, such as CD34, CD45, and CD133, may differ between the virtually same stem and progenitor cells, i.e., endothelial progenitor or mesenchymal stem cells, when they were obtained from different tissues. This finding raises questions whether phenotypic differences are due to the source or if it is only caused by different isolation and experimental conditions. PMID- 20024912 TI - Disruption of bis leads to the deterioration of the vascular niche for hematopoietic stem cells. AB - The stem cell niche plays an important role in the microenvironmental regulation of hematopoietic stem cells, but the integration of niche activity remains poorly understood. In this study, we show that a functional deficiency of Bis/BAG-3/CAIR 1, a protein related to apoptosis and the response to cellular stress, results in perturbation of the vascular stem cell niche, causing a series of hematopoietic derangements. Mice with a targeted disruption of bis (bis(-/-)) exhibited a loss of hematopoietic stem cells and defective B-cell development. However, this hematological defect of bis(-/-) mice was not reproduced when bis(-/-) bone marrow cells were transplanted into bis(+/+) recipients. Moreover, bis(+/+) bone marrow cells, when transplanted into bis(-/-) mice, reproduced the same defect as bis(-/-) cells, pointing to the microenvironmental origin of the phenotypes. Subsequent analysis of bis(-/-) mice bone marrow revealed a characteristic defect in the vascular stem cell niche that included the defective growth of stromal progenitor cells in colony forming unit-fibroblasts, the defect in sinusoidal endothelium, and the loss of stromal cells expressing CXCL-12 or IL-7 in the bone marrow. In contrast, no abnormalities were observed in the growth and hematopoietic supporting activities of osteoblasts from bis(-/-) mice bone marrows. Collectively, these results indicate that Bis functions to mediate cellular regulation of the stem cell niche on the vascular compartment and suggest that the vascular and osteoblastic compartments of the stem cell niche can be independently regulated during the in vivo orchestration of hematopoiesis. PMID- 20024913 TI - Improved peak capacity for CE separations of enzyme inhibitors with activity based detection using magnetic bead microreactors. AB - A technique for separating and detecting enzyme inhibitors was developed using CE with an enzyme microreactor. The on-column enzyme microreactor was constructed using NdFeB magnet(s) to immobilize alkaline phosphatase-coated superparamagnetic beads (2.8 microm diameter) inside a capillary before the detection window. Enzyme inhibition assays were performed by injecting a plug of inhibitor into a capillary filled with the substrate, AttoPhos. Product generated in the enzyme microreactor was detected by LIF. Inhibitor zones electrophoresed through the capillary, passed through the enzyme microreactor, and were observed as negative peaks due to decreased product formation. The goal of this study was to improve peak capacities for inhibitor separations relative to previous studies, which combined continuous engagement electrophoretically mediated microanalysis and transient engagement electrophoretically mediated microanalysis to study enzyme inhibition. The effects of electric field strength, bead injection time and inhibitor concentrations on peak capacity and peak width were investigated. Peak capacities were increased to >or=20 under optimal conditions of electric field strength and bead injection time for inhibition assays with arsenate and theophylline. Five reversible inhibitors of alkaline phosphatase (theophylline, vanadate, arsenate, L-tryptophan and tungstate) were separated and detected to demonstrate the ability of this technique to analyze complex inhibitor mixtures. PMID- 20024914 TI - A sensitive method for simultaneous determination of four macrolides by CE with electrochemiluminescence detection and its applications in human urine and tablets. AB - A sensitive method for simultaneous determination of azithromycin (AZI), acetylspiramycin (ACE), erythromycin (ERY), and josamycin (JOS) was developed by CE coupled with electrochemiluminescence detection with Ru(bpy)(3) (2+). The parameters related to separation and detection were investigated in detail. The four macrolides were well separated and detected within 6 min under the optimized conditions. The LOD (S/N=3) of AZI, ACE, ERY, and JOS were 1.2 x 10(-9), 7.1 x 10(-9), 3.9 x 10(-8) and 9.5 x 10(-8) mol/L, respectively. The LOQ (S/N=10) of AZI, ACE, ERY, and JOS in human urine were 8.2 x 10(-8), 2.5 x 10(-7), 8.9 x 10( 7) and 1.2 x 10(-6) mol/L, respectively. The recoveries of the four macrolides in human urine and pharmaceutical tablet samples were 85.0-104.0% at different concentration levels. PMID- 20024915 TI - Testing of the influenza virus purification by CIEF. AB - In virological practice, the pre-concentration, purification and subsequent determination of the purity and concentration of the viruses from the cultural medium and/or from the real sample are required. The conventional techniques used today are equipment demanding, time-consuming and laborious. In this study, the CIEF of influenza viruses with UV detection has been developed and subsequently used to test the purification of the virus from the biological samples. The equine and swine influenza viruses present in infected allantoic fluid of specific pathogen free embryonated chicken eggs were precipitated by using PEG 6000 and sodium chloride. The precipitated viruses were centrifuged at 14 000 x g, and the impurities of different densities were removed by using the sucrose gradients. The efficiency of the virus purification technique was examined by the CIEF and compared to the results of real-time PCR. The pIs of both influenza viruses were determined. Simultaneously, the CIEF was found to be a suitable method for the rapid testing of the efficiency of the virus purification. PMID- 20024916 TI - CE-ESI-TOF/MS for human growth hormone analysis. AB - CE is a powerful analytical tool used to separate intact biomolecules such as proteins. The coupling of CE with TOF/MS produces a very promising method that can be used to detect and identify proteins in different matrices. This paper describes an efficient, rapid, and simple CE-ESI-TOF/MS procedure for the analysis of endogenous human growth hormone and recombinant human growth hormone without sample preparation. Operational factors were optimized using an experimental design, and the method was successfully applied to distinguish human growth hormone and recombinant human growth hormone in unknown samples. PMID- 20024917 TI - Monodispersed submicron porous silica particles functionalized with CD derivatives for chiral CEC. AB - Rapid and efficient enantioseparation of halogen aryl alcohols and beta-blockers propranolol and pindolol in packed bed CEC (p-CEC) using as-prepared submicron porous silica chiral stationary phases (CSPs) has been achieved. Monodispersed 0.66 and 0.81 microm chiral submicron porous silica spheres were prepared using tetramethoxysilane and hexadecyltrimethylammonium bromide, followed by a hydrothermal treatment method with ammonia-ethanol to expand the pore of silica spheres without changing their spherical morphology. A proper specific surface of ca. 230 m(2)/g and pore sizes average of 6-8 nm were obtained by this method. The submicron porous silica spheres were modified with mono-6-phenylcarbamoylated beta-CD via thiol-en radical addition. They were packed into 9 cm 50 microm id capillary columns with photopolymerized monolithic frits. These submicron CSPs showed greater column efficiency (about 476 000 plates/m for 4-iodophenyl-1 ethanol) and higher resolution than the corresponding 3 microm CSP. PMID- 20024918 TI - Continuous intact cell detection and viability determination by CE with dual wavelength detection. AB - We introduce here a method for continuous intact cell detection and viability determination of individual trypan blue stained cells by CE with ultraviolet visible dual-wavelength detection. To avoid cell aggregation or damage during electrophoresis, cells after staining were fixed with 4% formaldehyde and were continuously introduced into the capillary by EOF. The absorbance of a cell at 590 nm was used to determine its viability. An absorbance of two milli-absorbance unit at 590 nm was the clear cut-off point for living and dead Hela cells in our experiments. Good viability correlation between the conventional trypan blue staining assay and our established CE method (correlation coefficient, R(2)=0.9623) was demonstrated by analysis of cell mixtures with varying proportions of living and dead cells. The CE method was also used to analyze the cytotoxicity of methylmercury, and the results were in good agreement with the trypan blue staining assay and 3-(4,5-dimethyl-2-thiazyl)-2,5-diphenyl-2H tetrazolium bromide methods. Compared with the 3-(4,5-dimethyl-2-thiazyl)-2,5 diphenyl-2H-tetrazolium bromide method, our established CE method can be easily automated to report cell viability based on the state of individual cells. Tedious manual cell counting and human error due to investigator bias can be avoided by using this method. PMID- 20024919 TI - Sonographic diagnosis of penile Mondor's disease associated with absence of a dorsal penile artery. AB - Penile Mondor's disease is an unusual condition, which is characterized by thrombosis in superficial dorsal penile vein. Here, we report the sonographic findings of this disease in a 35-year-old male along with the absence of right dorsal penile artery. PMID- 20024920 TI - An electrophoretic method for the detection of chymotrypsin and trypsin activity directly in whole blood. AB - In biomedical research and clinical diagnostics, it is a major challenge to measure disease-related degradative enzyme activity directly in whole blood. Present techniques for assaying degradative enzyme activity require sample preparation, which makes the assays time-consuming and costly. This study now describes a simple and rapid electrophoretic method that allows detection of degradative enzyme activity directly in whole blood using charge-changing fluorescent peptide substrates. Charge-changing substrates eliminate the need for sample preparation by producing positively charged cleavage fragments that can be readily separated from the oppositely charged fluorescent substrate and blood components by electrophoresis. Two peptide substrates have been developed for pancreatic alpha-chymotrypsin and trypsin. For the first substrate, a detection limit of 3 ng for both alpha-chymotrypsin and trypsin was achieved in whole rat blood using a 4% agarose gel. This substrate had minimal cross-reactivity with the trypsin-like proteases thrombin, plasmin, and kallikrein. For the second substrate (trypsin-specific), a detection limit of about 10-20 pg was achieved using thinner higher resolution 20 and 25% polyacrylamide gels. Thus, the new charge changing peptide substrates enable a simple electrophoretic assay format for the measurement of degradative enzyme activity, which is an important step toward the development of novel point-of-care diagnostics. PMID- 20024921 TI - Sonographic diagnosis of postpartum pseudoaneurysms of the uterine artery: a report of 2 cases. AB - Two cases of post-partum uterine artery pseudoaneurysm are described, 1 after normal vaginal delivery and the other after Caesarean section. Both cases were complicated by heavy bleeding and treated with catheter embolization. A turbulent arterial-type velocity waveform with high peak velocity and forward end-diastolic flow was registered in the feeding vessel. The peak blood flow velocity was high in both cases. The resistance index in the uterine artery was lower in the affected side of the uterus. Three-dimensional angiosonography was used to calculate the volume of the lesion, demonstrate its shape, and identify the feeding vessel. PMID- 20024922 TI - Reflections on Takahama Jr. et al's "Comparison of two prognostic scores for patients with parotid carcinoma". PMID- 20024923 TI - Study on enantiomeric separation of basic drugs by NACE in methanol-based medium using erythromycin lactobionate as a chiral selector. AB - A wide variety of chiral selectors have been employed in CZE, and among them macrocyclic antibiotics including glycopeptides, ansamycins, aminoglycosides and polypeptides exhibited prominent enantioselective properties toward abundant racemic compounds. Compared with CZE, the use of macrocyclic antibiotics as chiral selectors in NACE has not been reported previously. In this study, an approach to the enantioseparation of basic drugs by means of NACE with erythromycin lactobionate (EL) belonging to the group of macrolide antibiotics has been investigated. Especially different from the above four classes of antibiotics, there are no reports concerned with the use of macrolides which belong to macrocyclic antibiotics as chiral selectors in CE. In this work EL is first used as a chiral selector in NACE for the enantiomeric separations of two racemic basic drugs that possess high separability consisting of propranolol and duloxetine. Furthermore, EL possesses advantages such as high solubility and low viscosity in the solvent and very weak UV absorption. The chiral separations were achieved using Tris-boric acid as the BGE and methanol as the organic medium. In the course of this work we observed that both migration time and enantioseparation were influenced by several parameters such as the pH and composition of the BGE, EL concentration, capillary temperature and applied voltage. Consequently, these parameters were systematically optimized in order to obtain the optimum enantioseparations. PMID- 20024924 TI - A framework for the development of STR genotyping in domestic animal species: characterization and population study of 12 canine X-chromosome loci. AB - This study reports the methodology used to search, select and characterize STR loci on the canine X chromosome using publicly available genome resources and following the current guidelines for human and non-human forensic testing. After several rounds of selection, 12 X-STR markers were optimized for simultaneous co amplification in a single PCR, and genetic profiles were determined in a sample of 103 unrelated dogs. Mendelian inheritance was verified and mutation rates were assessed using family groups. Alleles that varied in size were sequenced to create a standardized nomenclature proposal based on the number of repeats. All loci conformed to Hardy-Weinberg expectations. The resulting panel showed high forensic efficiency, presenting high values of power of discrimination (in males and females) and mean exclusion chance, both in trios involving female offspring and in duos composed of dam and male offspring. Its use may complement the information obtained by autosomal STR analysis and contribute to the resolution of complex cases of kinship in dogs. The presented methodology for the de novo construction of an STR multiplex may also provide a helpful framework for analogous work in other animal species. As an increasing number of reference genomes become available, convenient tools for individual identification and parentage testing based on STR loci selected from autosomes or sex chromosomes' sequences may be created following this strategy. PMID- 20024925 TI - Ghrelin-liposome interactions: characterization of liposomal formulations of an acylated 28-amino acid peptide using CE. AB - Ghrelin is a pharmacologically interesting peptide hormone due to its effects on appetite and metabolism. The cationic, octanoylated 28 amino acid peptide has a short biological half-life; thus, prolonged release formulations are of interest. Acylated peptides have been suggested to bind to or be incorporated into liposomes. Formulations based on neutral dipalmitoylphosphatidylcholine (DPPC) liposomes and phosphatidylcholine:cholesterol (70:30 mol%) liposomes, and negatively charged dipalmitoylphosphatidylcholine:dipalmitoylphosphatidylserine (DPPC:DPPS) (70:30 mol%) liposomes (2 mM total lipid concentration) were characterized using ACE. Pre-equilibrium CZE and frontal analysis CE methods circumventing capillary wall adsorption of the peptide and the liposomes and suitable for characterizing ghrelin-liposome interactions were developed. The cationic peptide exhibited low affinity (<10% bound) for DPPC and phosphatidylcholine:cholesterol (70:30 mol%) liposomes whereas electrostatic interactions caused a higher affinity for DPPC:DPPS (70:30 mol%) liposomes. Studies on desacyl ghrelin instead of ghrelin demonstrated the significance of the n-octanoyl side chain as an affinity providing moiety towards DPPC:DPPS liposomes (48 and 73% bound peptide, respectively). CE experiments showed that the binding was characterized by rapid dissociation kinetics. PMID- 20024926 TI - High-throughput negative detection of SDS-PAGE separated proteins and its application for proteomics. AB - A negative detection method for proteins on SDS-PAGE is described. In this method, Eosin Y (EY) was selectively precipitated in the gel background, which is absent from those zones where proteins are located through the formation of a stable water-soluble protein-dye complex. Negative staining of proteins using EY, allows high-sensitivity, low-cost, and simple protocol. The new described method takes less than an hour to complete all the protocol, with a detection limit of 0.5 ng of single protein band. Comparing with imidazole-zinc negative stain, EY dye provides broader linear dynamic range, higher sensitivity and reproducibility, and better obvious contrast between the protein bands or spots and background. Furthermore, the novel technique developed here presented a real practical method for simultaneous processing of multiple gels, which makes it possible to perform high-throughput staining for proteome research. Additionally, we have also compared the influence of staining method on the quality of mass spectra by PMF. PMID- 20024927 TI - Ultrasonic-assisted derivatization of estrogenic compounds in a cup horn booster and determination by GC-MS. AB - Cup horn boosters are miniaturized ultrasound baths that maximize efficiency and precision. The optimization of an ultrasonic-assisted derivatization step by means of a cup horn booster and the determination of estrone, 17beta-estradiol, estriol, 17alpha-ethynyl estradiol and mestranol was developed by GC-MS. Different derivatization reagents and solvents were studied for maximizing the di derivatization of 17alpha-ethynyl estradiol under ultrasound energy. Only N,O bis(trimethylsilyl)trifluoroacetamide with 1% of trimethylchlorosilane in pyridine gave satisfactory results and this mixture was further used in the optimization of the ultrasound assisted derivatization. The experiment designs included sonication time (1-10 min), sonication power (20-80%), sonication cycles (1-9), derivatization reagent volume (25-125 microL) and solvent volume (25-125 microL). Once the optimum conditions were fixed, the effect of organic matter and the frequency of the water bath change were studied. Finally, the validation of the analytical method was carried out using spiked natural and synthetic waters. Recoveries (natural (138-70%) and synthetic (112-89%)), the LODs (0.35-1.66 ng/L), and LOQs (1.16-5.52 ng/L) and the precision (0.2-5.3%) of the method were studied. This is the first work in the literature where a cup horn booster is used with the aim of minimizing derivatization time during the determination of estrogenic compounds. PMID- 20024928 TI - Study of mass transfer in a dynamic hollow-fibre liquid phase microextraction system. AB - The extraction characteristics of a dynamic hollow-fibre liquid phase microextraction system were investigated by studying the mass transfer and diffusion rates of dinitrophenols from plasma samples over the liquid membrane (dihexylether). The measured diffusion coefficients were compared with theoretical values calculated from Stokes diameters. The diffusion mechanism was simulated by computer and the most polar compounds, 2,4-dinitrophenol and 4,6-o dinitrocresol, had associated diffusion coefficients that were close to the calculated theoretical values. 2-sec-Butyl-4,6 dinitrophenol and 2-tert-butyl-4,6 dinitrophenol, the compounds with the highest log P values, were retained by the polypropylene membrane, which reduced the experimentally observed diffusion rates to about half of the theoretical values. The retention was most likely due to dispersive forces interacting with the pore inner walls. Extraction was linearly correlated with time for all compounds and the repeatability was high (RSDs 7 11%), even for the shortest extraction times. Method LOD as the amount injected ranged between 0.3 and 3.1 ng for an extraction cycle of 213 s. PMID- 20024929 TI - LC of sulfonamide residues in poultry muscle and eggs extracts using fluorescence pre-column derivatization and monolithic silica column. AB - A new, rapid, sensitive and selective HPLC method with fluorescence detection is described for the simultaneous determination of 12 sulfonamides, in the presence of putrescine as internal standard, after pre-column derivatization with fluorescamine. The drugs were separated on a Chromolith Performance RP-18 column (100x4.6 mm), using a gradient elution with a binary mobile phase of methanol/0.05 M acetate buffer (pH 3.4). Linearity of derivatization was obtained for concentrations from 3.0 to 300 microg/L in standard solutions. The whole procedure was evaluated and fully validated, according to the European Union Decision 2002/657/EC, for the determination of sulfonamides in turkey muscle and hen eggs following SPE. The LODs varied from 2 to 17 microg/kg in turkey and 2 to 15 microg/kg in egg samples. The average recoveries ranged between 96.9-108.6% in turkey muscle and 96.0-108.4% in egg samples, respectively. PMID- 20024930 TI - 2-D hydrophilic interaction liquid chromatography-RP separation in urinary proteomics--minimizing variability through improved downstream workflow compatibility. AB - Optimization of every step in a bottom-up urinary proteomics approach was studied with respect to maximize the protein recovery and making the downstream steps in the workflow fully compatible without compromising on the amount of information obtained. Sample enrichment and desalting using centrifugal filtration (5 kDa cut off) yielded protein recoveries up to 97% when 8 M urea was used. Although yielding lower recoveries (88%), addition of Tris-HCl/NaCl was considered a better choice due to good down-stream compatibility. The consecutive depletion of HSA, using an immunoaffinity column was successfully adapted for use in urine. Separation of the trypsin generated peptides in an off-line 2-D chromatographic system consisting of a hydrophilic interaction liquid chromatography column, followed by a RP chromatography column showed a high peak capacity and good repeatability in addition to a high degree of orthogonality. All operations were modified in order to keep sample handling between every step to a minimum, reducing the variability of each process. In order to test the suitability of the full method in an extensive proteomic experiment, a urine sample from a kidney transplanted patient was analyzed (n=6). The total variability of the method was identified with RSD values ranging from 11 to 30%. Eventually, we identified a total of 1668 peptides and 438 proteins from a single urine sample despite the use of low-resolution MS/MS equipment. The optimized and "streamlined" complex method has shown potential for use in future urinary proteomic studies. PMID- 20024931 TI - Testing computational toxicology models with phytochemicals. AB - Computational toxicology employing quantitative structure-activity relationship (QSAR) modeling is an evidence-based predictive method being evaluated by regulatory agencies for risk assessment and scientific decision support for toxicological endpoints of interest such as rodent carcinogenicity. Computational toxicology is being tested for its usefulness to support the safety assessment of drug-related substances (e.g. active pharmaceutical ingredients, metabolites, impurities), indirect food additives, and other applied uses of value for protecting public health including safety assessment of environmental chemicals. The specific use of QSAR as a chemoinformatic tool for estimating the rodent carcinogenic potential of phytochemicals present in botanicals, herbs, and natural dietary sources is investigated here by an external validation study, which is the most stringent scientific method of measuring predictive performance. The external validation statistics for predicting rodent carcinogenicity of 43 phytochemicals, using two computational software programs evaluated at the FDA, are discussed. One software program showed very good performance for predicting non-carcinogens (high specificity), but both exhibited poor performance in predicting carcinogens (sensitivity), which is consistent with the design of the models. When predictions were considered in combination with each other rather than based on any one software, the performance for sensitivity was enhanced, However, Chi-square values indicated that the overall predictive performance decreases when using the two computational programs with this particular data set. This study suggests that complementary multiple computational toxicology software need to be carefully selected to improve global QSAR predictions for this complex toxicological endpoint. PMID- 20024932 TI - Toxicology and risk assessment of coumarin: focus on human data. AB - Coumarin is a secondary phytochemical with hepatotoxic and carcinogenic properties. For the carcinogenic effect, a genotoxic mechanism was considered possible, but was discounted by the European Food Safety Authority in 2004 based on new evidence. This allowed the derivation of a tolerable daily intake (TDI) for the first time, and a value of 0.1 mg/kg body weight was arrived at based on animal hepatotoxicity data. However, clinical data on hepatotoxicity from patients treated with coumarin as medicinal drug is also available. This data revealed a subgroup of the human population being more susceptible for the hepatotoxic effect than the animal species investigated. The cause of the high susceptibility is currently unknown; possible mechanisms are discussed. Using the human data, a TDI of 0.1 mg/kg body weight was derived, confirming that of the European Food Safety Authority. Nutritional exposure may be considerably, and is mainly due to use of cassia cinnamon, which is a popular spice especially, used for cookies and sweet dishes. To estimate exposure to coumarin during the Christmas season in Germany, a telephone survey was performed with more than 1000 randomly selected persons. Heavy consumers of cassia cinnamon may reach a daily coumarin intake corresponding to the TDI. PMID- 20024933 TI - Antioxidative and antigenotoxic properties of vegetables and dietary phytochemicals: the value of genomics biomarkers in molecular epidemiology. AB - There is considerable evidence that consumption of fruits and vegetables may contribute to the prevention of cancer. It is however remarkable that evidence for such a preventive action arising from mechanistic studies is becoming stronger, whereas results of some recent prospective studies are less convincing. This apparent discrepancy may be overcome, or at least understood, by introducing molecular markers in future epidemiological studies, taking modulation of molecular processes as well as genetic variability in human populations into account. Both human and animal studies demonstrated that vegetable intake modulates gene expression in the gastrointestinal tract of many genes involved in biological pathways in favor of cancer risk prevention. Gene sets identified in this type of studies can be further evaluated, linked to the biological effects of phytochemicals and developed into biomarkers for larger human studies. Human dietary intervention studies have demonstrated that, apart from target tissues, also peripheral lymphocytes can be used for biomonitoring of chemopreventive effects. Transcriptomic responses and metabolite profiling may link phenotypic markers of preventive effects to specific molecular processes. The use of genomics techniques appears to be a promising approach to establish mechanistic pathways involved in chemoprevention by phytochemicals, particularly when genetic variability is taken into account. PMID- 20024934 TI - Involvement of ERK, Akt and JNK signalling in H2O2-induced cell injury and protection by hydroxytyrosol and its metabolite homovanillic alcohol. AB - The olive oil polyphenol, hydroxytyrosol (HT), is believed to be capable of exerting protection against oxidative kidney injury. In this study we have investigated the ability of HT and its O-methylated metabolite, homovanillic alcohol (HVA) to protect renal cells against oxidative damage induced by hydrogen peroxide. We show that both compounds were capable of inhibiting hydrogen peroxide-induced kidney cell injury via an ability to interact with both MAP kinase and PI3 kinase signalling pathways, albeit at different concentrations. HT strongly inhibited death and prevented peroxide-induced increases in ERK1/2 and JNK1/2/3 phosphorylation at 0.3 microM, whilst HVA was effective at 10 microM. At similar concentrations, both compounds also prevented peroxide-induced reductions in Akt phosphorylation. We suggest that one potential protective effect exerted by olive oil polyphenols against oxidative kidney cell injury may be attributed to the interactions of HT and HVA with these important intracellular signalling pathways. PMID- 20024935 TI - Transcriptome analysis in benefit-risk assessment of micronutrients and bioactive food components. AB - The establishment of functional effects due to variation in concentrations of micronutrients in our diet is difficult since they are often not immediately recognized as being healthy or unhealthy. Indeed, effects induced by micronutrients are hard to identify and therefore the establishment of the recommended daily intake, the optimal intake and the upper limit pose a challenge. For bioactive food components this is even more complicated. Whole genome transcriptome analysis is highly suitable to obtain unbiased information on potential affected biological processes on a whole-genome level. Here, we will describe and discuss several aspects of transcriptome analysis in benefit-risk assessment, including effect size, sensitivity and statistical power, that have to be taken into account to faithfully identify functional effects of micronutrients and bioactive food components. PMID- 20024936 TI - Guidelines for LC-MS identifications of flavoring substances in nature, made by the Working Group on Methods of Analysis of the International Organization of the Flavor Industry (IOFI). AB - These guidelines are intended to assist authors and reviewers of publications concerning the validity of identifications of flavouring substances in nature using LC-MS. They address the choice of the transition between precursor and product ions (with examples using the "Identification Point" concept), the tolerances for ion-abundance ratios, and the importance of performing blank experiments. PMID- 20024938 TI - Identifying functional promoter SNPs using allelic imbalance. PMID- 20024939 TI - Database overkill. PMID- 20024941 TI - dbSNP in the detail and copy number complexities. AB - dbSNP is a general catalog of genetic polymorphism maintained by NCBI, mainly collating information for single nucleotide variations, many of which will be single nucleotide polymorphisms (SNPs), but also including small indels. It takes submissions from many sources, now also including large numbers of sequence variants identified by next-generation sequencing. A number of differently designed studies have attempted to estimate the error rates in data archived in dbSNP. Most recently, a study added to earlier studies identifying specific issues for duplicons and copy number variations (CNVs); earlier analyses have focused on stop codons, splice sites, and the general content of dbSNP. This article overviews dbSNP itself, these studies, and their implications. PMID- 20024942 TI - Cystic papillary thyroid carcinoma in fine needle aspiration may represent a subset of the encapsulated variant in WHO classification. AB - This study audits the reliability of ultrasound-guided fine needle aspiration (FNA) in excluding papillary thyroid carcinoma (PTC) in thyroid cysts containing mural nodules, and investigates the histological counterpart of cystic PTC diagnosed on FNA. Using a 10-5 MHz ultrasound probe and a 27-gauge needle, solid portions of thyroid nodules were sampled and assessed immediately using both Diff Quik and Ultrafast Papanicolaou stains. Unlike usual PTCs that demonstrate hypercellularity, the aspirates of cystic PTC showed low cellularity and consisted of papillae with atypical nuclei scattered in abundant thin colloid. Over a period of 13 years, histological follow-up was obtained from 11 women and 6 men in whom cystic PTC was reported on FNA. This represented 4.4% of 383 cases of PTC reported and 0.25% of all thyroid FNAs performed. In all 17 cases, histopathology showed encapsulated PTC in various stages of cystic degeneration. Sonography correlated well with histopathology, where findings ranged from cysts with small mural nodules to solid nodules with pockets of thin colloid. In 87 patients with thyroid cysts containing mural nodules, FNA findings were benign, as was clinical follow-up that ranged from 1 to 12 years. In conclusion, ultrasound-guided FNA, if performed in the manner described, can reliably distinguish cystic PTC from a benign cyst with a mural nodule. Cystic PTC on FNA in this series correlates to a subset of the encapsulated variant of PTC, an entity described in the 1988 WHO Histological Typing of Thyroid Tumours in the good prognostic category. PMID- 20024940 TI - Testosterone metabolites differentially maintain adult morphology in a sexually dimorphic neuromuscular system. AB - The lumbar spinal cord of rats contains the sexually dimorphic, steroid-sensitive spinal nucleus of the bulbocavernosus (SNB). Androgens are necessary for the development of the SNB neuromuscular system, and in adulthood, continue to influence the morphology and function of the motoneurons and their target musculature. However, estrogens are also involved in the development of the SNB system, and are capable of maintaining function in adulthood. In this experiment, we assessed the ability of testosterone metabolites, estrogens and nonaromatizable androgens, to maintain neuromuscular morphology in adulthood. Motoneuron and muscle morphology was assessed in adult normal males, sham castrated males, castrated males treated with testosterone, dihydrotestosterone, estradiol, or left untreated, and gonadally intact males treated with the 5alpha reductase inhibitor finasteride or the aromatase inhibitor fadrozole. After 6 weeks of treatment, SNB motoneurons were retrogradely labeled with cholera toxin HRP and reconstructed in three dimensions. Castration resulted in reductions in SNB target muscle size, soma size, and dendritic morphology. Testosterone treatment after castration maintained SNB soma size, dendritic morphology, and elevated target muscle size; dihydrotestosterone treatment also maintained SNB dendritic length, but was less effective than testosterone in maintaining both SNB soma size and target muscle weight. Treatment of intact males with finasteride or fadrozole did not alter the morphology of SNB motoneurons or their target muscles. In contrast, estradiol treatment was completely ineffective in preventing castration-induced atrophy of the SNB neuromuscular system. Together, these results suggest that the maintenance of adult motoneuron or muscle morphology is strictly mediated by androgens. PMID- 20024943 TI - Additive survival least-squares support vector machines. AB - This work studies a new survival modeling technique based on least-squares support vector machines. We propose the use of a least-squares support vector machine combining ranking and regression. The advantage of this kernel-based model is threefold: (i) the problem formulation is convex and can be solved conveniently by a linear system; (ii) non-linearity is introduced by using kernels, componentwise kernels in particular are useful to obtain interpretable results; and (iii) introduction of ranking constraints makes it possible to handle censored data. In an experimental setup, the model is used as a preprocessing step for the standard Cox proportional hazard regression by estimating the functional forms of the covariates. The proposed model was compared with different survival models from the literature on the clinical German Breast Cancer Study Group data and on the high-dimensional Norway/Stanford Breast Cancer Data set. PMID- 20024944 TI - Structural neuroplasticity in the sensorimotor network of professional female ballet dancers. AB - Evidence suggests that motor, sensory, and cognitive training modulates brain structures involved in a specific practice. Functional neuroimaging revealed key brain structures involved in dancing such as the putamen and the premotor cortex. Intensive ballet dance training was expected to modulate the structures of the sensorimotor network, for example, the putamen, premotor cortex, supplementary motor area (SMA), and the corticospinal tracts. We investigated gray (GM) and white matter (WM) volumes, fractional anisotropy (FA), and mean diffusivity (MD) using magnetic resonance-based morphometry and diffusion tensor imaging in 10 professional female ballet dancers compared with 10 nondancers. In dancers compared with nondancers, decreased GM volumes were observed in the left premotor cortex, SMA, putamen, and superior frontal gyrus, and decreased WM volumes in both corticospinal tracts, both internal capsules, corpus callosum, and left anterior cingulum. FA was lower in the WM underlying the dancers' left and right premotor cortex. There were no significant differences in MD between the groups. Age of dance commencement was negatively correlated with GM and WM volume in the right premotor cortex and internal capsule, respectively, and positively correlated with WM volume in the left precentral gyrus and corpus callosum. Results were not influenced by the significantly lower body mass index of the dancers. The present findings complement the results of functional imaging studies in experts that revealed reduced neural activity in skilled compared with nonskilled subjects. Reductions in brain activity are accompanied by local decreases in GM and WM volumes and decreased FA. PMID- 20024945 TI - Lateral line system and its innervation in Tetraodontiformes with outgroup comparisons: descriptions and phylogenetic implications. AB - The lateral line system and its innervation in ten tetraodontiform families and five outgroup taxa were examined. Although some homology issues remained unresolved, tetraodontiforms were characterized by having two types (at least) of superficial neuromasts (defined by the presence or absence of supporting structures) and accessory lateral lines and neuromasts (except Molidae in which "accessory" elements were absent). The preopercular line in Tetraodontiformes was not homologous with that of typical teleosts, because the line was innervated by the opercular ramule that was newly derived from the mandibular ramus, the condition being identical to that in Lophiidae. Within Tetraodontiformes, the number of neuromasts varied between 70 and 277 in the main lines and between 0 and 52 in accessory elements. Variations were also recognized in the presence or absence of the supraorbital commissure, mandibular line, otic line, postotic line, ventral trunk line, and some lateral line nerve rami, most notably the dorsal branch of the opercular ramule, being absent in Aracanidae, Ostraciidae, Tetraodontidae, Diodontidae, and Molidae. Morphological characteristics derived from the lateral line system and its innervation provided some support for a sister relationship of tetraodontiforms with lophiiforms. PMID- 20024946 TI - Comparative myoanatomy of cycliophoran life cycle stages. AB - The metazoan phylum Cycliophora includes small cryptic epibionts that live attached to the mouthparts of clawed lobsters. The life cycle is complex, with alternating sexual and asexual generations, and involves several sessile and free living stages. So far, the morphological and genetic characterization of cycliophorans has been unable to clarify the phylogenetic position of the phylum. In this study, we add new details on the muscular anatomy of the feeding stage, the attached Prometheus larva, the dwarf male, and the female of one of the two hitherto described species, Symbion pandora. The musculature of the feeding stage is composed of myofibers that run longitudinally in the buccal funnel (two fibers) and in the trunk (variable number of fibers). The mouth opening is lined by a myoepithelial ring musculature. A complex myoepithelial sphincter is situated proximal to the anus. In the attached Prometheus larva, three longitudinal sets of myofilaments run dorsally, laterally, and ventrally along the entire anterior-posterior body axis. The muscular architecture of the dwarf male is complex, especially close to the penis, in the posterior part of the body. An X-shaped muscle structure is found on the dorsal side, whereas on the ventral side, longitudinal muscles and a V-shaped muscle structure are present. These muscles are complemented by additional dorsoventral muscles. The mesodermal muscle fibers attach to the cuticle via the epidermis in all life cycle stages studied herein. The musculature of the female is similar to that of the Pandora larva of Symbion americanus and includes dorsoventral muscles and longitudinal muscles that run in the dorsal and ventral body region. Overall, our results reveal striking similarities in the muscular arrangement of the life cycle stages of both Symbion species. PMID- 20024947 TI - Impact of Bacillus thuringiensis strains on survival, reproduction and foraging behaviour in bumblebees (Bombus terrestris). AB - BACKGROUND: Bacillus thuringiensis (Bt) and its protein crystals are used worldwide, either as a spray or when expressed in transgenic crops, for the control of pest insects. However, owing to their intensive use, there exists a debate regarding the involvement of this microbial insecticide in bee colony losses. In this study, in a tiered approach using laboratory microcolonies, an evaluation was made of the potential lethal and sublethal hazards on colony reproduction and foraging behaviour of workers of the bumblebee Bombus terrestris (L.) of two commercial Bt strains: kurstaki (Dipel) and aizawai (Xentari). Bumblebees, like honey bees, are intensively used in modern agriculture for pollination and fulfil a crucial role in the natural ecosystem. RESULTS: Exposure of bumblebees dermally or via treated pollen to either of the two Bt formulations at their field recommended rates (0.1%) caused no reduction in survival. However, when applied in the feeding sugar water, aizawai killed all workers at a concentration of 0.1%, but this lethal effect was lost at 0.01%. With respect to reproductive effects, kurstaki was harmless, while aizawai at 0.1% delivered in the feeding sugar water and pollen reduced reproduction by 100 and 31% respectively. Lower doses of 0.01% aizawai in the sugar water showed no more effect. In addition, kurstaki at 0.1% and aizawai at 0.01% in the feeding sugar water did not impair the foraging behaviour, resulting in normal nest colony performance. CONCLUSION: The results with kurstaki and aizawai demonstrated that, in general, the Bt strains are safe to B. terrestris bumblebees, although in some cases there were detrimental effects that depended on strain and route of exposure. In addition, the authors believe that to draw firm conclusions regarding the hazards of Bt to bumblebees would require more information on relevant concentrations of Bt products in the environment. Hence, routine testing for lethal and sublethal effects is recommended to ascertain combined use of Bt products and bumblebees in modern agriculture. PMID- 20024948 TI - Managing earthworm casts (Oligochaeta: Lumbricidae) in turfgrass using a natural byproduct of tea oil (Camellia sp.) manufacture. AB - BACKGROUND: Earthworm casts are a worldwide problem on golf courses and sports fields when they disrupt the playability, aesthetics and maintenance of closely mowed playing surfaces. Currently, no pesticides are labeled for earthworms in the United States. Tea seed pellets (TSPs), a saponin-rich byproduct of Camellia oleifera Abel oil manufacture, were tested for expelling earthworms and reducing casts on creeping bentgrass turf. The fate of expelled worms, methods for removing them and impacts on pest and beneficial arthropods were also evaluated. RESULTS: Application of TSPs at 2.93 kg 100 m(-2), followed by irrigation, quickly expelled earthworms from the soil. A single application reduced casts by 80-95% for at least 5 weeks. Mowing or sweeping removed expelled earthworms from putting green surfaces. Most expelled earthworms burrowed down when transferred to untreated turf, but few survived. Bioassay-guided fractionation confirmed the vermicidal activity results from a mix of saponins. TSPs did not reduce the abundance of beneficial soil arthropods, nor did they control black cutworms or white grubs in treated turf. CONCLUSION: TSPs are an effective botanical vermicide that could be useful for selectively managing earthworm casts on closely mowed turfgrass. They might also be used to suppress earthworms in grassy strips alongside runways to reduce bird strike hazard at airports. PMID- 20024949 TI - Characterisation and toxicity of Bacillus thuringiensis strains from hazelnut pests and fields. AB - BACKGROUND: In order to find and identify more toxic insecticidal Bacillus thuringiensis Berliner (Bt) strains, a survey was carried out of B. thuringiensis isolate pests belonging to Coleoptera, Lepidoptera and Diptera and from soils in hazelnut fields. Of 16 isolates having Bacillus cereus-B. thuringiensis morphology, eight were classified as B. thuringiensis because of the production of parasporal delta-endotoxin crystals. RESULTS: In this study, eight isolates of B. thuringiensis from hazelnut pests (isolates Bn1, Mm2, Mnd and Xd3) and from hazelnut soils (isolates 6, 27, 40 and 46) have been characterised in detail. These isolates were compared with reference strains by electron microscopy, SDS PAGE analysis, cry gene content, serological test and insecticidal activity. CONCLUSION: Results indicate that Bn1 and MnD are B. thuringiensis subsp. kurstaki, and Mm2 and Xd3 are B. thuringiensis subsp. tenebrionis. In addition, isolate 6 is B. thuringiensis subsp. israelensis, isolates 27 and 46 are B. thuringiensis subsp. kumamotoensis and isolate 40 is B. thuringiensis subsp. indiana. The four B. thuringiensis isolates from hazelnut pests may be valuable as biological control agents against coleopteran and lepidopteran insects. PMID- 20024950 TI - Combined QF-PCR and MLPA molecular analysis of miscarriage products: an efficient and robust alternative to karyotype analysis. AB - OBJECTIVES: To replace G-banded chromosome analysis for miscarriage products with a combined molecular approach: QF-PCR and MLPA, to increase efficiency, reduce costs, and improve the diagnostic success rate for these samples. METHODS: A review of 10 years of karyotype results for miscarriages products indicated that 2.7% of nonmosaic chromosome imbalance would not be detected by the molecular approach. The molecular approach was validated on 117 samples in parallel with karyotype analysis; no discrepancies were detected. The molecular approach was implemented in September 2007, and in the first 18 months 500 samples were processed. RESULTS: In 500 samples, 117 samples (23%) were abnormal. Of these abnormalities, 64% were trisomies, 12% triploid, 11% monosomy X and 13% other abnormalities. When compared to karyotype analysis, the success rate was higher (95% cf 70%) and the reporting time was lower (88% within 28 days cf 79%). In addition, efficiency was higher as labour-intensive cell culture and karyotyping were replaced by batch testing and automated analysis. CONCLUSIONS: This molecular approach is less labour-intensive, allows a higher sample throughput and has a higher success rate than karyotype analysis; it is therefore an efficient and cost-effective diagnostic testing strategy for miscarriage products. PMID- 20024951 TI - NMR parameters of two tricyclododecadienes--strained hydrocarbons with close distance between perpendicularly or parallelly arranged double bonds. AB - Quantum chemical calculations of the hypothetical tricyclo[5.5.0.0(4, 10)]dodeca 1(7),4(10)-diene with a perpendicular arrangement of double bonds and of the known tricyclo[4.2.2.2(2, 5)]dodeca-1,5-diene isomer with the parallel arrangement yield, in agreement with expectations, greater stability of the latter molecule. They reproduce the available experimental results for chemical shifts and coupling constants for the latter molecule. Large calculated values of sigma(Csp2) and those of (1)J(C=C) for both dienes are not due to nonplanar distortions on the double bonds but due to the close distances between them. The calculated NMR parameters of the hypothetical diene could be useful for its future identification as the calculated values of the parameters for 1 are larger than those for 2. PMID- 20024952 TI - Theoretical and experimental evaluation of continuous arterial spin labeling techniques. AB - Continuous arterial spin labeling is known to be the most sensitive arterial spin labeling technique. To avoid magnetization transfer effects and to overcome hardware limitations, several sequences have been proposed that adiabatically label the inflowing blood. Four of these methods are examined with respect to their sensitivity both theoretically by Bloch equation simulations and experimentally. All sequences were optimized carefully by adjusting their measurement parameters based exclusively on the results of simulations. Perfusion measurements on the human brain obtained at 3 T result in excellent images from all techniques, while differences in sensitivity are similar to those expected from the simulations. PMID- 20024953 TI - 4D phase contrast MRI at 3 T: effect of standard and blood-pool contrast agents on SNR, PC-MRA, and blood flow visualization. AB - Time-resolved phase contrast (PC) MRI with velocity encoding in three directions (flow-sensitive four-dimensional MRI) can be employed to assess three-dimensional blood flow in the entire aortic lumen within a single measurement. These data can be used not only for the visualization of blood flow but also to derive additional information on vascular geometry with three-dimensional PC MR angiography (MRA). As PC-MRA is sensitive to available signal-to-noise ratio, standard and novel blood pool contrast agents may help to enhance PC-MRA image quality. In a group of 30 healthy volunteers, the influence of different contrast agents on vascular signal-to-noise ratio, PC-MRA quality, and subsequent three dimensional stream-line visualization in the thoracic aorta was determined. Flow sensitive four-dimensional MRI data acquired with contrast agent provided significantly improved signal-to-noise ratio in magnitude data and noise reduction in velocity data compared to measurements without contrast media. The agreement of three-dimensional PC-MRA with reference standard contrast-enhanced MRA was good for both contrast agents, with improved PC-MRA performance for blood pool contrast agent, particularly for the smaller supra-aortic branches. For three-dimensional flow visualization, a trend toward improved results for the data with contrast agent was observed. PMID- 20024954 TI - The effects of osmotic stress on the structure and function of the cell nucleus. AB - Osmotic stress is a potent regulator of the normal function of cells that are exposed to osmotically active environments under physiologic or pathologic conditions. The ability of cells to alter gene expression and metabolic activity in response to changes in the osmotic environment provides an additional regulatory mechanism for a diverse array of tissues and organs in the human body. In addition to the activation of various osmotically- or volume-activated ion channels, osmotic stress may also act on the genome via a direct biophysical pathway. Changes in extracellular osmolality alter cell volume, and therefore, the concentration of intracellular macromolecules. In turn, intracellular macromolecule concentration is a key physical parameter affecting the spatial organization and pressurization of the nucleus. Hyper-osmotic stress shrinks the nucleus and causes it to assume a convoluted shape, whereas hypo-osmotic stress swells the nucleus to a size that is limited by stretch of the nuclear lamina and induces a smooth, round shape of the nucleus. These behaviors are consistent with a model of the nucleus as a charged core/shell structure pressurized by uneven partition of macromolecules between the nucleoplasm and the cytoplasm. These osmotically-induced alterations in the internal structure and arrangement of chromatin, as well as potential changes in the nuclear membrane and pores are hypothesized to influence gene transcription and/or nucleocytoplasmic transport. A further understanding of the biophysical and biochemical mechanisms involved in these processes would have important ramifications for a range of fields including differentiation, migration, mechanotransduction, DNA repair, and tumorigenesis. PMID- 20024955 TI - Preventive effect of (-)epigallocatechin gallate on lipids, lipoproteins, and enzymes of lipid metabolism in isoproterenol-induced myocardial infarction in rats. AB - This article reports data on the preventive effect of (-)epigallocatechin gallate (EGCG) on lipid metabolism and lipoproteins in isoproterenol (ISO)-induced myocardial infarction (MI) in Wistar rats. The rats were induced MI by ISO (100 mg/kg) at an interval of 24 h for 2 days. EGCG (30 mg/kg) was given to rats as pretreatment for 21 days orally using an intragastric tube. EGCG significantly reduced the increased serum levels of cholesterol, triglycerides, and free fatty acids in the heart and serum phospholipids (PLs) in ISO-treated rats. It also significantly increased the reduced levels of heart PLs in ISO-induced rats. EGCG reduced the levels of serum low-density lipoprotein cholesterol and very low density lipoprotein cholesterol and increased serum high-density lipoprotein (HDL)-cholesterol in ISO-treated rats. It also reduced the increased cholesterol/PL ratio and atherogenic index and significantly increased the reduced ratio of HDL-cholesterol/total cholesterol. Also EGCG significantly increased the reduced activity of lecithin cholesterol acyl transferase in ISO treated rats. Thus, EGCG prevented the accumulation of lipids and altered the levels of lipoproteins in myocardial-infarcted rats. PMID- 20024956 TI - Metabolism of the mesoionic compound (MI-D) by mouse liver microsome, detection of its metabolite in vivo, and acute toxicity in mice. AB - The mesoionic derivative 4-phenyl-5-[4-nitrocinnamoyl]-1,3,4-thiadiazolyl-2 phenylamine chloride (MI-D) has antitumoral and anti-inflammatory effects. In this study, we present aspects of its metabolism and toxicity in mice. MI-D was metabolized in vitro by liver microsome, generating a main product with a much shorter retention time than MI-D in high-performance liquid chromatography (HPLC) analysis but with a spectrum similar to that of the original molecule. Mass spectrometry with electrospray ionization in positive mode analysis of the purified compound by HPLC indicated that the product of metabolism has four additional hydroxyl groups (m/z = 465) compared with MI-D (m/z = 401). The HPLC analyses of plasma and urine samples from mice treated with MI-D showed the presence of the metabolite product. The kinetic parameters K(m) (19.5 +/- 4.5 microM) and V(max) [1.5 +/- 0.4 units of fluorescence/(100 microg of microsomal protein/mL/s)] were estimated, confirming the metabolism of MI-D and indicating that the reaction follows Michaelis-Menten kinetics. Acute toxicity was established on the basis of an estimation of mean lethal dose (LD-50; 181.2 mg/kg) and histopathological analysis of animals that survived the LD-50 test. Abdominal adhesions, inflammatory foci, and formation of granulomas were observed. Altogether, the results contribute to the advancement of research in support of MI-D as a future chemotherapeutic drug. PMID- 20024957 TI - Oncogene expression profiles in K6/ODC mouse skin and papillomas following a chronic exposure to monomethylarsonous acid. AB - We have previously observed that a chronic drinking water exposure to monomethylarsonous acid [MMA(III)], a cellular metabolite of inorganic arsenic, increases tumor frequency in the skin of keratin VI/ornithine decarboxylase (K6/ODC) transgenic mice. To characterize gene expression profiles predictive of MMA(III) exposure and mode of action of carcinogenesis, skin and papilloma RNA was isolated from K6/ODC mice administered 0, 10, 50, and 100 ppm MMA(III) in their drinking water for 26 weeks. Following RNA processing, the resulting cRNA samples were hybridized to Affymetrix Mouse Genome 430A 2.0 GeneChips(R). Micoarray data were normalized using MAS 5.0 software, and statistically significant genes were determined using a regularized t-test. Significant changes in bZIP transcription factors, MAP kinase signaling, chromatin remodeling, and lipid metabolism gene transcripts were observed following MMA(III) exposure as determined using the Database for Annotation, Visualization and Integrated Discovery 2.1 (DAVID) (Dennis et al., Genome Biol 2003;4(5):P3). MMA(III) also caused dose-dependent changes in multiple Rho guanine nucleotide triphosphatase (GTPase) and cell cycle related genes as determined by linear regression analyses. Observed increases in transcript abundance of Fosl1, Myc, and Rac1 oncogenes in mouse skin support previous reports on the inducibility of these oncogenes in response to arsenic and support the relevance of these genomic changes in skin tumor induction in the K6/ODC mouse model. PMID- 20024958 TI - Sex-specific effects of gestational and lactational coexposure to lead and cadmium on hepatic phase I and phase II xenobiotic/steroid-metabolizing enzymes and antioxidant status. AB - Liver has evolved complex enzymatic mechanisms to detoxify a wide array of xenobiotic substances, ranging from dietary components to environmental toxins to pharmaceuticals. Activities of many steroid-metabolizing enzymes in adult rat liver microsomes are sexually differentiated. Toxic effects of lead and cadmium on hepatic tissue have been well established in our earlier studies. We thus monitored the effects of gestational and lactational coexposure to lead and cadmium on hepatic phase I and phase II xenobiotic- and steroid-metabolizing enzyme activities in both male and female F1 generation postnatal day (PND) 56 rats. Adult pregnant female rats were treated subcutaneously [0.05 mg/(kg body wt. day)] with sodium acetate (control group), lead acetate, and cadmium acetate separately and in combination throughout the gestational and lactational period. Hepatic phase I xenobiotic-metabolizing enzymes (NADPH- and NADH-cytochrome c reductase) activities significantly decreased significantly in all the metal treated groups in both PND 56 male and female rats as compared with the control group. Hepatic phase II enzymes (uridine diphosphate-glucuronosyl transferase, gamma-glutamyl transpeptidase, glutathione-S-transferase, 17-beta-hydroxysteroid oxidoreductase) were also highly susceptible to all the metal-treated groups. The observed alterations in the oxidative stress and biochemical parameters in the liver of F1 generation male and female rats resulted from an independent effect of lead and/or cadmium and also from their interaction. Results suggest that early developmental exposure to lead and cadmium both alone and in combination can suppress the hepatic xenobiotic-metabolizing enzyme activities in the liver of F1 generation male and female rats in a sex-dependent manner. PMID- 20024959 TI - Mast cells promote atherosclerosis by inducing both an atherogenic lipid profile and vascular inflammation. AB - Accumulating in vitro and in vivo studies have proposed a role for mast cells in the pathogenesis of atherosclerosis. Here, we studied the role of mast cells in lipoprotein metabolism, a key element in the atherosclerotic disease. Male mice deficient in low-density lipoprotein receptors and mast cells on a Western diet for 26 weeks had significantly less atherosclerotic changes both in aortic sinus (55%, P = 0.0009) and in aorta (31%, P = 0.049), as compared to mast cell competent littermates. Mast cell-deficient female mice had significantly less atherosclerotic changes in aortic sinus (43%, P = 0.011). Furthermore, we found a significant positive correlation between the extent of atherosclerosis and the number of adventitial/perivascular mast cells in aortic sinus of mast cell competent mice (r = 0.615, P = 0.015). Serum cholesterol and triglyceride levels were significantly lower in both male (63%, P = 0.0005 and 57%, P = 0.004) and female (73%, P = 0.00009 and 54%, P = 0.007) mast cell-deficient mice, with a concomitant decrease in atherogenic apoB-containing particles and serum prebeta high-density lipoprotein and phospholipid transfer protein activity in both male (69% and 24%) and female (74% and 54%) mast cell-deficient mice. Serum soluble intercellular adhesion molecule was decreased in both male (32%, P = 0.004) and female (28%, P = 0.003) mast cell-deficient mice, whereas serum amyloid A was similar between mast cell-deficient and competent mice. In conclusion, mast cells participate in the pathogenesis of atherosclerosis in ldlr(-/-) mice by inducing both an atherogenic lipid profile and vascular inflammation. PMID- 20024960 TI - Mechanism of diepoxybutane-induced p53 regulation in human cells. AB - Diepoxybutane (DEB) is the most potent active metabolite of the environmental chemical 1,3-butadiene (BD). BD is a known mutagen and human carcinogen and possesses multisystems organ toxicity. We previously reported the elevation of p53 in human TK6 lymphoblasts undergoing DEB-induced apoptosis. In this study, we have characterized the DEB-induced p53 accumulation and investigated the mechanisms by which DEB regulates this p53 accumulation. The elevation of p53 levels in DEB-exposed TK6 lymphoblasts and human embryonic lung (HEL) human fibroblasts was found to be largely due to the stabilization of the p53 protein. DEB increased the acetylation of p53 at lys-382, dramatically reduced complex formation between p53 and its regulator protein mdm2 and induced the phosphorylation of p53 at serines 15, 20, 37, 46, and 392 in human lymphoblasts. A dramatic increase in phosphorylation of p53 at serine 15 in correlation to total p53 levels was observed in DEB-exposed Ataxia Telangiectasia Mutated (ATM) proficient human lymphoblasts as compared to DEB-exposed ATM-deficient human lymphoblasts; this implicates the ATM kinase in the elevation of p53 levels in DEB-exposed cells. Collectively, these findings explain for the first time the mechanism by which p53 accumulates in DEB-exposed cells and contributes to the understanding of the molecular toxicity of DEB and BD. PMID- 20024961 TI - Nutritional composition and digestibility by ocelots (Leopardus pardalis) of whole animals and a commercial diet. AB - Felids are adapted to eat whole prey, but in North American zoos are usually fed processed diets based on muscle meat. We analyzed proximate nutrient composition and digestibility by ocelots of a commercial processed diet and whole animals of five species. The processed diet did not differ significantly from the whole animals in proximate composition, although it was at one end of the range of results for all nutrients. Domestic chicks were significantly lower than all other dietary items tested in digestibility of energy and fat, and lower than rabbits and quail in digestibility of dry matter. There were no other significant differences. These results suggest that the commercial diet tested provides an appropriate nutritional environment for ocelots with respect to proximate constituents. Studies of vitamin and mineral composition and digestibility and comparisons to wild prey species should be conducted to permit a full evaluation. PMID- 20024962 TI - The potential for microchip-automated technology to improve enrichment practices. AB - The greatest limitation involved with providing enrichment and evaluating its effects on captive mammals is a lack of available staff time [Hoy et al., 2009]. One method to overcome this is through the use of automated husbandry systems to dispense enrichment to individual animals by using implantable radio frequency microchips. An international multi-institutional survey was conducted to determine whether the personnel involved with management, research, and husbandry of captive mammals would support the development of such automated husbandry systems. The survey was also designed to establish which applications of the system would be considered to be most beneficial. Overall, there was a very high level of interest by survey respondents in adopting automated husbandry systems and all potential applications were considered beneficial to varying degrees. These results, together with the relatively low cost of installation and operation, demonstrate that in the future automated enrichment could be an important and effective means of enhancing captive animal management. PMID- 20024964 TI - Osseointegration into a novel titanium foam implant in the distal femur of a rabbit. AB - A novel porous titanium foam implant has recently been developed to enhance biological fixation of orthopaedic implants to bone. The aim of this study was to examine the mechanical and histological characteristics of bone apposition into two different pore sizes of this titanium foam (565 and 464 micron mean void intercept length) and to compare these characteristics to those obtained with a fully porous conventionally sintered titanium bead implant. Cylindrical implants were studied in a rabbit distal femoral intramedullary osseointegration model at time zero and at 3, 6, and 12 weeks. The amount of bone ingrowth, amount of periprosthetic bone, and mineral apposition rate of periprosthetic bone measured did not differ among the three implant designs at 3, 6, or 12 weeks. By 12 weeks, the interface stiffness and maximum load of the beaded implant was significantly greater than either foam implant. No significant difference was found in the interface stiffness or maximum load between the two foam implant designs at 3, 6, or 12 weeks. The lower compressive modulus of the foam compared to the more dense sintered beaded implants likely contributed to the difference in failure mode. However, the foam implants have a similar compressive modulus to other clinically successful coatings, suggesting they are nonetheless clinically adequate. Additional studies are required to confirm this in weight-bearing models. Histological data suggest that these novel titanium foam implants are a promising alternative to current porous coatings and should be further investigated for clinical application in cementless joint replacement. PMID- 20024965 TI - Corrosion of phosphate-enriched titanium oxide surface dental implants (TiUnite) under in vitro inflammatory and hyperglycemic conditions. AB - Endosseous dental implants use is increasing in patients with systemic conditions that compromise wound healing. Manufacturers recently have redesigned implants to ensure more reliable and faster osseointegration. One design strategy has been to create a porous phosphate-enriched titanium oxide (TiUnite) surface to increase surface area and enhance interactions with bone. In the current study, the corrosion properties of TiUnite implants were studied in cultures of monocytic cells and solutions simulating inflammatory and hyperglycemic conditions. Furthermore, to investigate whether placement into bone causes enough mechanical damage to alter implant corrosion properties, the enhanced surface implants as well as machined titanium implants were placed into human cadaver mandibular bone, the bone removed, and the corrosion properties measured. Implant corrosion behavior was characterized by open circuit potentials, linear polarization resistance, and electrical impedance spectroscopy. In selected samples, THP1 cells were activated with lipopolysaccharide prior to implant exposure to simulate an inflammatory environment. No significant differences in corrosion potentials were measured between the TiUnite implants and the machined titanium implants in previous studies. TiUnite implants exhibited lower corrosion rates in all simulated conditions than observed in PBS, and EIS measurements revealed two time constants which shifted with protein-containing electrolytes. In addition, the TiUnite implants displayed a significantly lower corrosion rate than the machined titanium implants after placement into bone. The current study suggests that the corrosion risk of the enhanced oxide implant is lower than its machined surface titanium implant counterpart under simulated conditions of inflammation, elevated dextrose concentrations, and after implantation into bone. PMID- 20024966 TI - Comparing the efficacy of three bioceramic matrices for the release of vancomycin hydrochloride. AB - A number of calcium phosphate materials have been investigated as drug release matrices for the prophylactic treatment of implant-related osteomyelitis. However, some studies have shown the influence of processing on the efficacy of the delivered drug. The objective of this study was to evaluate the influence of pH during processing on the efficacy of vancomycin hydrochloride (VH) against Staphylococcus aureus. VH was loaded into a brushite cement (CaHPO(4).2H(2)O; pH 2.4); a hydroxyapatite cement (Ca(10)(PO(4))(6)OH(2); pH 9.4); and an apatite xerogel (pH 7.4). The pH of the material during processing had a significant influence on the mechanism of release from the cement. VH released from the apatite cement (pH 9.4) was not released in accordance with the Higuchi model. In addition to affecting release, the basic pH was shown to diminish the antibacterial potency of the released VH. Despite exceeding the minimum inhibitory concentration, the eluent from the apatite cement was ineffective against a culture of S. aureus. The findings of this study reinforce the importance of evaluating not only the release of the drug from the material matrix but also the antibacterial potency of the released drug. PMID- 20024967 TI - Intravaginal and in vitro temperature changes with tampons of differing composition and absorbency. AB - Vaginal tampons are Class II medical devices used by women to manage menstruation. The purpose of this study was to investigate intravaginal temperature changes with simulated and actual menstrual tampon use. Tampons (with varying absorbent compositions) embedded with a thermocouple sensor were used to study temperature effects in vitro in a model of the vagina (condom placed in a hollow glass tube, jacketed in a 37 degrees C water bath, and dosed with human menses to fluid saturation) and clinically during menstrual tampon wear under controlled conditions (up to 8 h in a stationary, supine position). Elevations in the temperature of the tampon core occurred upon menses fluid acquisition both in vitro and clinically. Temperature profile characteristics varied from a transient spike with commercial cotton-rayon blend tampons of two different absorbencies to a small but sustained rise (> or =6 h) with a carboxymethyl cellulose (CMC) containing prototype. On the basis of the results from this study, fluid absorption by tampons generates an exothermic event whose characteristics vary with tampon design and composition. We speculate the small, sustained increased in tampon temperature noted during this study may enhance the production of a bacterial exotoxin associated with tampons composed of CMC. PMID- 20024968 TI - Antibacterial effects of 4-META/MMA-TBB resin containing chlorhexidine. AB - This study evaluated the antibacterial effects of 4-acryloyloxyethyl trimellitate anhydride/methyl methacrylate-tri-n-butylborane (4-META/MMA-TBB) resin containing chlorhexidine (CHX) digluconate. The CHX was incorporated into the resins at varying concentrations from 0.0 (control) to 3.0%. The antibacterial effect of each resin against seven microorganisms was examined using agar diffusion tests. Growth inhibition of two streptococci was also tested in brain heart infusion (BHI) liquid broth containing each resin. Release of CHX was analyzed using HPLC, and enamel and dentin bond strengths of each resin were measured. In the agar diffusion tests, growth of all the microorganisms was inhibited by the resin specimens containing CHX at 1.0% or higher except for Enterococcus faecalis. There was greater sensitivity in Streptococcus mutans and Streptococcus sobrinus than in the other bacteria tested. Complete growth inhibition of these two streptococci was observed in BHI liquid broth containing 1.0-1.5% or greater CHX incorporated resin. According to HPLC analysis, 1.0% and 1.5% CHX-containing resins released means of 3.63 and 8.59 microg/mL of CHX, respectively. Specimens with 0.5-1.5% and 0.5-2.0% CHX exhibited no significant reduction in enamel and dentin bond strengths, respectively, when compared to the control (p > 0.05). This in vitro study suggested that incorporation of 1.0-1.5% CHX digluconate into the 4-META resin is optimal in terms of antibacterial effects and bond strength to the tooth. PMID- 20024969 TI - Fibrillogenesis in continuously spun synthetic collagen fiber. AB - The universal structural role of collagen fiber networks has motivated the development of collagen gels, films, coatings, injectables, and other formulations. However, reported synthetic collagen fiber fabrication schemes have either culminated in short, discontinuous fiber segments at unsuitably low production rates, or have incompletely replicated the internal fibrillar structure that dictates fiber mechanical and biological properties. We report a continuous extrusion system with an off-line phosphate buffer incubation step for the manufacture of synthetic collagen fiber. Fiber with a cross-section of 53+ or - 14 by 21 + or - 3 microm and an ultimate tensile strength of 94 + or - 19 MPa was continuously produced at 60 m/hr from an ultrafiltered monomeric collagen solution. The effect of collagen solution concentration, flow rate, and spinneret size on fiber size was investigated. The fiber was further characterized by microdifferential scanning calorimetry, transmission electron microscopy (TEM), second harmonic generation (SHG) analysis, and in a subcutaneous murine implant model. Calorimetry demonstrated stabilization of the collagen triple helical structure, while TEM and SHG revealed a dense, axially aligned D-periodic fibril structure throughout the fiber cross-section. Implantation of glutaraldehyde crosslinked and noncrosslinked fiber in the subcutaneous tissue of mice demonstrated limited inflammatory response and biodegradation after a 6-week implant period. PMID- 20024971 TI - Elucidation of real-time hardening mechanisms of two novel high-strength calcium phosphate bone cements. AB - Despite the numerous literature data available in the field of calcium phosphate bone cements, the mechanism and kinetics of their hardening, both of which are of great importance for cements application, in most cases, is unknown. In this work, the mechanism and kinetics of hardening of two novel high-strength calcium phosphate bone cements were studied using the energy dispersive X-ray diffraction technique, which allows rapid collection of the patterns. The phase transformations occurring on the setting and hardening processes were monitored in situ. Containing minimal quantity of components, whose mixing leads to the formation of cements with pH close to neutral, the cements under study are simple in handling. The main component of both formulations is tetracalcium phosphate. In both cements, the effect of the addition of high- and low-molecular weight chitosan on phase development and kinetics was investigated in detail. One of the cements has the compressive strength of about 70 MPa, whereas the strength of the other, containing Ca(3)Al(2)O(6), is much higher, about 100 MPa. This latter cement could be regarded as an alternative to the common low-strength bioresorbable brushite cements. PMID- 20024970 TI - Molecular and tissue responses in the healing of rat calvarial defects after local application of simvastatin combined with alpha tricalcium phosphate. AB - We have previously reported that healing of rat calvarial defects was enhanced by application of alpha tricalcium phosphate (alphaTCP) combined with simvastatin, a cholesterol synthesis inhibitor. The purpose of the present study was to investigate the cellular and molecular mechanisms in this phenomenon. Rat calvarial defects were grafted with alphaTCP with or without simvastatin or left untreated. Animals were sacrificed on 3, 7, 10, 14, and 21 days postoperatively and histological changes in the defect region were assessed. Gene expression patterns were examined by RT-PCR. Proliferation and migration of osteoprogenitor cells from the dura mater were increased in simvastatin group from day 3 to day 10 (p < 0.01). New bone formation was significantly increased in simvastatin group on day 14 and day 21 (p < 0.01). BMP-2 expression was significantly higher in simvastatin group on day 3 and day 14 (p < 0.05) and maintained until day 21. Increased upregulation of TGF-beta1 was also observed in the simvastatin group on day 7 (p < 0.05) which was maintained until day 14. These findings suggest that the proliferation and recruitment of osteoprogenitor cells were critical steps in early stage of bone healing and that these steps were enhanced by TGF-beta1 and BMP-2, which were stimulated by simvastatin. PMID- 20024972 TI - DNA microarrays to define and search for genes associated with obesity. AB - One of the major goals of this review was to identify obesity-specific gene profiles in animal models to help comprehend the pathogenic mechanisms and the prediction of the phenotypic outcomes of obesity and its associated metabolic diseases. The genomic examination of insulin-sensitive tissues, such as the adipose and hepatic tissues, has provided a wealth of information about the changes in gene expression in obesity and its associated metabolic diseases. The overexpression of genes related to inflammation, immune response, adhesion molecules, and lipid metabolism is a major characteristic of white adipose tissue, while the overexpression of the genes related to lipid metabolism, adipocyte differentiation, defense, and stress responses is noticeable in the non alcoholic fatty liver of obese rodents. The hepatic-gene expression profiles led us to hypothesize that in obese rodents, the livers are supplied with large amounts of free fatty acids under conditions associated with obesity either through increased fatty acid biosynthesis or through decreased fatty acid oxidation, which may lead to increased mitochondrial respiratory activity. The wide list of genes that were identified in previous studies could be a source of potential therapeutic targets because most of these genes are involved in the key mechanisms of obesity development, from adipocyte differentiation to the disturbance of metabolism. PMID- 20024973 TI - Structural features of apramycin bound at the bacterial ribosome a site as detected by NMR and CD spectroscopy. PMID- 20024975 TI - Concentration-dependent hydrogen-bonding effects on the dimethyl sulfoxide vibrational structure in the presence of water, methanol, and ethanol. AB - The effects of hydrogen bonding between dimethyl sulfoxide (DMSO) and the co solvents water, methanol, and ethanol on the symmetric and antisymmetric CSC stretching vibrations of DMSO are investigated by means of Raman spectroscopy. The Raman spectra are recorded as a function of co-solvent concentration and reflect changes in structure and polarizability as well as hydrogen-bond donor and acceptor ability. In all cases studied a nonideal mixing behavior is observed. The spectra of the DMSO/water system show blue-shifted CSC stretching modes. The antisymmetric frequencies are always further blue-shifted than the symmetric stretching ones. The DMSO/methanol system also features blue-shifted CSC stretching frequencies but at high mole fractions a pronounced red shifting is observed. In the binary DMSO/ethanol system, the co-solvent also gives rise to blue shifts of the CSC stretching frequencies but restricted to mole fractions between x=0.38 and 0.45. The different magnitudes and occurrences of both blue- and red-shifted spectral lines are comprehensively and critically discussed with respect to the existing literature concerning wavenumbers and Raman intensities in both absolute and normalized values. In particular, the normalized Raman intensities show a higher sensitivity for the nonideal mixing behavior because they are independent of the mole fraction. PMID- 20024976 TI - A molecular magnet confined in the nanocage of a globular protein. AB - The effect of confinement and energy transfer on the dynamics of a molecular magnet, known as a model system to study quantum coherence, is investigated. For this purpose the well-known polyoxovanadate [V(15)As(6)O(42)(H(2)O)](6-) (V(15)) is incorporated into a protein (human serum albumin, HSA) cavity. Due to a huge overlap of the optical absorption spectrum of V(15) with the emission spectrum of a fluorescence center of HSA (containing a single tryptophan residue), energy transfer is induced and probed by steady-state and time-resolved fluorescence. The geometrical coordination and the distance of the confined V(15) to the tryptophan moiety of HSA are investigated at various temperatures. This effect is used as a local probe for the thermal denaturation of the protein at elevated temperatures. PMID- 20024977 TI - Nano-ice on boron nitride nanomesh: accessing proton disorder. AB - Water was investigated on a h-BN/Rh(111) nanomesh template using variable temperature scanning tunneling microscopy (STM) and density functional theory (DFT) calculations. Below 52 K, two distinct phases self-assemble within the 3.2 nm unit cell of the nanomesh that consists of "holes" and "wires". In the 2 nm holes, an ordered phase of nano-ice crystals with about 40 molecules is found. The ice crystals arrange in a bilayer honeycomb lattice, where hydrogen atoms of the lower layer point to the substrate. The phase on the 1 nm wires is a low density gas phase. Tunneling barrier height dI/dz spectroscopy measurements reveal the dipoles of individual molecules in the nano-ice clusters and access proton disorder. PMID- 20024978 TI - Rational design of amyloid binding agents based on the molecular rotor motif. PMID- 20024980 TI - Conversion of cellulose to hexitols catalyzed by ionic liquid-stabilized ruthenium nanoparticles and a reversible binding agent. PMID- 20024979 TI - 5-Benzylidenethiazolidin-4-ones as multitarget inhibitors of bacterial Mur ligases. AB - Mur ligases participate in the intracellular path of bacterial peptidoglycan biosynthesis and constitute attractive, although so far underexploited, targets for antibacterial drug discovery. A series of hydroxy-substituted 5 benzylidenethiazolidin-4-ones were synthesized and tested as inhibitors of Mur ligases. The most potent compound 5 a was active against MurD-F with IC(50) values between 2 and 6 microm, making it a promising multitarget inhibitor of Mur ligases. Antibacterial activity against different strains, inhibitory activity against protein kinases, mutagenicity and genotoxicity of 5 a were also investigated, and kinetic and NMR studies were conducted. PMID- 20024981 TI - 3-Heterocycle-phenyl N-alkylcarbamates as FAAH inhibitors: design, synthesis and 3D-QSAR studies. AB - Carbamates are a well-established class of fatty acid amide hydrolase (FAAH) inhibitors. Here we describe the synthesis of meta-substituted phenolic N alkyl/aryl carbamates and their in vitro FAAH inhibitory activities. The most potent compound, 3-(oxazol-2yl)phenyl cyclohexylcarbamate (2 a), inhibited FAAH with a sub-nanomolar IC(50) value (IC(50)=0.74 nM). Additionally, we developed and validated three-dimensional quantitative structure-activity relationships (QSAR) models of FAAH inhibition combining the newly disclosed carbamates with our previously published inhibitors to give a total set of 99 compounds. Prior to 3D-QSAR modeling, the degree of correlation between FAAH inhibition and in silico reactivity was also established. Both 3D-QSAR methods used, CoMSIA and GRID/GOLPE, produced statistically significant models with coefficient of correlation for external prediction (R(2) (PRED)) values of 0.732 and 0.760, respectively. These models could be of high value in further FAAH inhibitor design. PMID- 20024982 TI - Thermally stable porous hydrogen-bonded coordination networks displaying dual properties of robustness and dynamics upon guest uptake. AB - Two series of microporous lanthanide coordination networks of the general formula, {[Ln(ntb)Cl(3)] x xH(2)O}(n) (series 1: monoclinic C2/c, Ln = Sm and Tb; series 2: hexagonal P3(1)/c, Ln = Sm and Eu; ntb = tris(benzimidazol-2 ylmethyl)amine, x = 0-4) have been synthesized and characterized by IR, elemental analyses, thermal gravimetry, and single-crystal and powder X-ray diffraction methods. In both series, the monomeric [Ln(ntb)Cl(3)] coordination units are consolidated by N-H...Cl or C-H...Cl hydrogen bonds to sustain three-dimensional (3D) networks. However, the different modes of hydrogen bonding in the two series lead to crystallization of the same [Ln(ntb)Cl(3)] monomers in different forms (monoclinic vs. hexagonal), consequently giving rise to distinct porous structures. The resulting hydrogen-bonded coordination networks display high thermal stability and robustness in water removal/inclusion processes, which was confirmed by temperature-dependent single-crystal-to-single-crystal transformation measurements. Adsorption studies with H(2), CO(2), and MeOH have been carried out, and reveal distinct differences in adsorption behavior between the two forms. In the case of MeOH uptake, the monoclinic network shows a normal type I isotherm, whereas the hexagonal network displays dynamic porous properties. PMID- 20024983 TI - Sigma-alkylpalladium intermediates in intramolecular Heck reactions:isolation and catalytic activity. AB - The isolation of sigma-alkylpalladium Heck intermediates, possible when beta hydride elimination is inhibited, is a rather rare event. Performing intramolecular Heck reactions on N-allyl-2-halobenzylamines in the presence of [Pd(PPh(3))(4)], we isolated and characterized a series of stable bridged palladacycles containing an iodine or bromine atom on the palladium atom. Indolyl substrates were also tested for isolation of the corresponding complexes. X-ray crystallographic analysis of one of the indolyl derivatives revealed the presence of a five-membered palladacycle with the metal center bearing a PPh(3) ligand and an iodine atom in a cis position with respect to the nitrogen atom. The stability of the sigma-alkylpalladium complexes is probably a consequence of the strong constraint resulting from the bridged junction that hampers the cisoid conformation essential for beta-hydride elimination. Subsequently, the thus obtained bridged five-membered palladacycles were proven to be effective precatalysts in Heck reactions as well as in cross-coupling processes such as Suzuki and Stille reactions. PMID- 20024984 TI - Pincer-type Heck catalysts and mechanisms based on Pd(IV) intermediates: a computational study. AB - Pincer-type palladium complexes are among the most active Heck catalysts. Due to their exceptionally high thermal stability and the fact that they contain Pd(II) centers, controversial Pd(II)/Pd(IV) cycles have been often proposed as potential catalytic mechanisms. However, pincer-type Pd(IV) intermediates have never been experimentally observed, and computational studies to support the proposed Pd(II)/Pd(IV) mechanisms with pincer-type catalysts have never been carried out. In this computational study the feasibility of potential catalytic cycles involving Pd(IV) intermediates was explored. Density functional calculations were performed on experimentally applied aminophosphine-, phosphine-, and phosphite based pincer-type Heck catalysts with styrene and phenyl bromide as substrates and (E)-stilbene as coupling product. The potential-energy surfaces were calculated in dimethylformamide (DMF) as solvent and demonstrate that Pd(II)/Pd(IV) mechanisms are thermally accessible and thus a true alternative to formation of palladium nanoparticles. Initial reaction steps of the lowest energy path of the catalytic cycle of the Heck reaction include dissociation of the chloride ligands from the neutral pincer complexes [{2,6 C(6)H(3)(XPR(2))(2)}Pd(Cl)] [X=NH, R=piperidinyl (1 a); X=O, R=piperidinyl (1 b); X=O, R=iPr (1 c); X=CH(2), R=iPr (1 d)] to yield cationic, three-coordinate, T shaped 14e(-) palladium intermediates of type [{2,6-C(6)H(3)(XPR(2))(2)}Pd](+) (2). An alternative reaction path to generate complexes of type 2 (relevant for electron-poor pincer complexes) includes initial coordination of styrene to 1 to yield styrene adducts [{2,6-C(6)H(3)(XPR(2))(2)}Pd(Cl)(CH(2)=CHPh)] (4) and consecutive dissociation of the chloride ligand to yield cationic square-planar styrene complexes [{2,6-C(6)H(3)(XPR(2))(2)}Pd(CH(2)=CHPh)](+) (6) and styrene. Cationic styrene adducts of type 6 were additionally found to be the resting states of the catalytic reaction. However, oxidative addition of phenyl bromide to 2 result in pentacoordinate Pd(IV) complexes of type [{2,6 C(6)H(3)(XPR(2))(2)}Pd(Br)(C(6)H(5))](+) (11), which subsequently coordinate styrene (in trans position relative to the phenyl unit of the pincer cores) to yield hexacoordinate phenyl styrene complexes [{2,6 C(6)H(3)(XPR(2))(2)}Pd(Br)(C(6)H(5))(CH(2)=CHPh)](+) (12). Migration of the phenyl ligand to the olefinic bond gives cationic, pentacoordinate phenylethenyl complexes [{2,6-C(6)H(3)(XPR(2))(2)}Pd(Br)(CHPhCH(2)Ph)](+) (13). Subsequent beta hydride elimination induces direct HBr liberation to yield cationic, square planar (E)-stilbene complexes with general formula [{2,6 C(6)H(3)(XPR(2))(2)}Pd(CHPh=CHPh)](+) (14). Subsequent liberation of (E)-stilbene closes the catalytic cycle. PMID- 20024985 TI - Substrate-controlled diastereoselectivity switch in catalytic asymmetric direct Mannich reaction of glycine derivatives with imines: from anti- to syn-alpha,beta diamino acids. PMID- 20024986 TI - Symmetrization of cationic hydrogen bridges of protonated sponges induced by solvent and counteranion interactions as revealed by NMR spectroscopy. AB - The properties of the intramolecular hydrogen bonds of doubly (15)N-labeled protonated sponges of the 1,8-bis(dimethylamino)naphthalene (DMANH(+)) type have been studied as a function of the solvent, counteranion, and temperature using low-temperature NMR spectroscopy. Information about the hydrogen-bond symmetries was obtained by the analysis of the chemical shifts delta(H) and delta(N) and the scalar coupling constants J(N,N), J(N,H), J(H,N) of the (15)NH(15)N hydrogen bonds. Whereas the individual couplings J(N,H) and J(H,N) were averaged by a fast intramolecular proton tautomerism between two forms, it is shown that the sum |J(N,H)+J(H,N)| generally represents a measure of the hydrogen-bond strength in a similar way to delta(H) and J(N,N). The NMR spectroscopic parameters of DMANH(+) and of 4-nitro-DMANH(+) are independent of the anion in the case of CD(3)CN, which indicates ion-pair dissociation in this solvent. By contrast, studies using CD(2)Cl(2), [D(8)]toluene as well as the freon mixture CDF(3)/CDF(2)Cl, which is liquid down to 100 K, revealed an influence of temperature and of the counteranions. Whereas a small counteranion such as trifluoroacetate perturbed the hydrogen bond, the large noncoordinating anion tetrakis[3,5 bis(trifluoromethyl)phenyl]borate B[{C(6)H(3)(CF(3))(2)}(4)](-) (BARF(-)), which exhibits a delocalized charge, made the hydrogen bond more symmetric. Lowering the temperature led to a similar symmetrization, an effect that is discussed in terms of solvent ordering at low temperature and differential solvent order/disorder at high temperatures. By contrast, toluene molecules that are ordered around the cation led to typical high-field shifts of the hydrogen-bonded proton as well as of those bound to carbon, an effect that is absent in the case of neutral NHN chelates. PMID- 20024987 TI - Systematic investigation of photoinduced electron transfer controlled by internal charge transfer and its consequences for selective PdCl2 coordination. AB - Fluoroionophores of fluorophore-spacer-receptor format were prepared for detection of PdCl(2) by fluorescence enhancement. The fluorescent probes 1-13 consist of a fluorophore group, an alkyl spacer and a dithiomaleonitrile PdCl(2) receptor. First, varying the length of the alkylene spacer (compounds 1-3) revealed a dominant through-space pathway for oxidative photoinduced electron transfer (PET) in CH(2)-bridged dithiomaleonitrile fluoroionophores. Second, fluorescent probes 4-9 containing two anthracene or pyrene fragments connected through CH(2) bridges to the dithiomaleonitrile unit were synthesized. Modulation of the oxidation potential (E(Ox)) through electron-withdrawing or -donating groups on the anthracene moiety regulates the thermodynamic driving force for oxidative PET (DeltaG(PET)) in bis(anthrylmethylthio)maleonitriles and therefore the fluorescence quantum yields (Phi(f)), too. The new concept was confirmed and transferred to pyrenyl ligands, and fluorescence enhancements (FE) greater than 3.2 in the presence of PdCl(2) were achieved by 7 and 8 (FE=5.4 and 5.2). Finally, for comparison, monofluorophore ligands 10-13 were synthesized. PMID- 20024988 TI - Enhancing the orienting properties of poly(gamma-benzyl-L-glutamate) by means of additives. AB - Residual dipolar couplings (RDCs) have recently become increasingly important in organic structure determination due to their unique information content. One main limitation for the use of RDCs in organic compounds is the orientation that needs to be induced to be able to measure RDCs. So far, there are very few possibilities to modulate the orientational properties of organic solutes and even less when chiral media are considered. Based on our recent findings that the critical concentration of the liquid-crystalline phase of homopolypeptides depends on their molecular weight, we sought for further ways to modulate the orienting properties. We were especially interested in seeing whether we could not only influence the induced degree of orientation, but whether we could also change the solute's preferred orientation and even enhance enantiodifferentiation. We thus tried different aprotic and protic additives and were successful in all of the above-mentioned aspects by using CCl(4) as the additive. Furthermore, we consider DMSO to be a very useful additive. The LC phase of low MW poly(gamma-benzyl-L-glutamate) (PBLG) is usually unstable when DMSO is added. The high MW PBLG used in this study, however, remained stable up to a DMSO/CDCl(3) ratio of 1:2. By using this combination of solvents, the alignment of the two enantiomers of a compound, which is insoluble in CDCl(3), namely, the HCl salt of a tryptophane ester, was possible leading to high-quality spectra. The two enantiomers of the tryptophane ester showed different couplings, thus indicating that enantiodifferentiation is taking place. Thus we were able to modulate the orienting properties (degree of orientation, preferred orientation and enantiodifferentiation) of PBLG by using additives and to increase the accessible solvent and solute range significantly. PMID- 20024989 TI - Efficient intramolecular charge transfer in oligoyne-linked donor-pi-acceptor molecules. AB - Studies are reported on a series of triphenylamine-(C[triple bond]C)(n)-2,5 diphenyl-1,3,4-oxadiazole dyad molecules (n=1-4, 1, 2, 3 and 4, respectively) and the related triphenylamine-C(6)H(4)-(C[triple bond]C)(3)-oxadiazole dyad 5. The oligoyne-linked D-pi-A (D=electron donor, A=electron acceptor) dyad systems have been synthesised by palladium-catalysed cross-coupling of terminal alkynyl and butadiynyl synthons with the corresponding bromoalkynyl moieties. Cyclic voltammetric studies reveal a reduction in the HOMO-LUMO gap in the series of compounds 1-4 as the oligoyne chain length increases, which is consistent with extended conjugation through the elongated bridges. Photophysical studies provide new insights into conjugative effects in oligoyne molecular wires. In non-polar solvents the emission from these dyad systems has two different origins: a locally excited (LE) state, which is responsible for a pi*-->pi fluorescence, and an intramolecular charge transfer (ICT) state, which produces charge-transfer emission. In polar solvents the LE state emission vanishes and only ICT emission is observed. This emission displays strong solvatochromism and analysis according to the Lippert-Mataga-Oshika formalism shows significant ICT for all the luminescent compounds with high efficiency even for the longer more conjugated systems. The excited-state properties of the dyads in non-polar solvents vary with the extent of conjugation. For more conjugated systems a fast non-radiative route dominates the excited-state decay and follows the Engelman-Jortner energy gap law. The data suggest that the non-radiative decay is driven by the weak coupling limit. PMID- 20024990 TI - Yb(OTf)3-catalyzed construction of indole derivatives through formal [3+3] cycloaddition of 1,1-vinylidenecyclopropanediesters with nitrones. PMID- 20024991 TI - Ring-opening/ring-closing protocols from nitrothiophenes: six-membered versus unusual eight-membered sulfur heterocycles through Michael-type addition on nitrobutadienes. AB - When Ar is a low-aromaticity homo- or heterosystem, the sulfonyl-stabilized anion of nitrobutadienes 4 (which derive from the initial ring opening of 3 nitrothiophene) undergoes a rather surprising addition onto the aromatic ring itself, thereby leading to the construction of an unusual eight-membered sulfur heterocycle condensed with the original Ar ring. The competitiveness of such a pathway with respect to the formation of the thiopyran ring (i.e., addition onto the nitrovinyl moiety) is favored at low temperatures, thus revealing its nature as a kinetically controlled process. PMID- 20024992 TI - A highly versatile catalyst system for the cross-coupling of aryl chlorides and amines. AB - The syntheses of 2-(di-tert-butylphosphino)-N,N-dimethylaniline (L1, 71%) and 2 (di-1-adamantylphosphino)-N,N-dimethylaniline (L2, 74 %), and their application in Buchwald-Hartwig amination, are reported. In combination with [Pd(allyl)Cl](2) or [Pd(cinnamyl)Cl](2), these structurally simple and air-stable P,N ligands enable the cross-coupling of aryl and heteroaryl chlorides, including those bearing as substituents enolizable ketones, ethers, esters, carboxylic acids, phenols, alcohols, olefins, amides, and halogens, to a diverse range of amine and related substrates that includes primary alkyl- and arylamines, cyclic and acyclic secondary amines, N-H imines, hydrazones, lithium amide, and ammonia. In many cases, the reactions can be performed at low catalyst loadings (0.5-0.02 mol % Pd) with excellent functional group tolerance and chemoselectivity. Examples of cross-coupling reactions involving 1,4-bromochlorobenzene and iodobenzene are also reported. Under similar conditions, inferior catalytic performance was achieved when using Pd(OAc)(2), PdCl(2), [PdCl(2)(cod)] (cod = 1,5 cyclooctadiene), [PdCl(2)(MeCN)(2)], or [Pd(2)(dba)(3)] (dba = dibenzylideneacetone) in combination with L1 or L2, or by use of [Pd(allyl)Cl](2) or [Pd(cinnamyl)Cl](2) with variants of L1 and L2 bearing less basic or less sterically demanding substituents on phosphorus or lacking an ortho-dimethylamino fragment. Given current limitations associated with established ligand classes with regard to maintaining high activity across the diverse possible range of C-N coupling applications, L1 and L2 represent unusually versatile ligand systems for the cross-coupling of aryl chlorides and amines. PMID- 20024993 TI - Direct growth of single-crystal Pt nanowires on Sn@CNT Nanocable: 3D electrodes for highly active electrocatalysts. AB - A newly designed and fabricated novel three dimensional (3D) nanocomposite composed of single-crystal Pt nanowires (PtNW) and a coaxial nanocable support consisting of a tin nanowire and a carbon nanotube (Sn@CNT) is reported. This nanocomposite is fabricated by the synthesis of Sn@CNT nanocables by means of a thermal evaporation method, followed by the direct growth with PtNWs through a facile aqueous solution approach at room temperature. Electrochemical measurements demonstrate that the PtNW--Sn@CNT 3D electrode exhibits enhanced electrocatalytic performance in oxygen reduction reaction (ORR) for polymer electrolyte membrane fuel cells (PEMFCs), methanol oxidation (MOR) for direct methanol fuel cells (DMFCs), and CO tolerance compared with commercial ETEK Pt/C catalyst made of Pt nanoparticles. PMID- 20024994 TI - Intermediates involved in the oxidation of nitrogen monoxide: photochemistry of the cis-N(2)O(2)O(2) complex and of sym-N(2)O(4) in solid Ne matrices. AB - Pure sym-N(2)O(4) isolated in solid Ne was obtained by passing cold neon gas over solid N(2)O(4) at -115 degrees C and quenching the resulting gaseous mixture at 6.3 K. Filtered UV irradiation (260-400 nm) converts sym-N(2)O(4) into trans ONONO(2), a weakly interacting (NO(2))(2) radical pair, and traces of the cis N(2)O(2)O(2) complex. Besides the weakly bound ONO(2) complex, cis-N(2)O(2)O(2) was also obtained by co-deposition of NO and O(2) in solid Ne at 6.3 K, and both complexes were characterised by their matrix IR spectra. Concomitantly formed cis N(2)O(2) dissociated on exposure to filtered IR irradiation (400-8000 cm(-1)), and the cis-N(2)O(2)O(2) complex rearranged to sym-N(2)O(4) and trans-ONONO(2). Experiments using (18)O(2) in place of (16)O(2) revealed a non-concerted conversion of cis-N(2)O(2)O(2) into these species, and gave access to four selectively di-(18)O-substituted trans-ONONO(2) isotopomers. No isotopic scrambling occurred. The IR spectra of sym-N(2)O(4) and of trans-ONONO(2) in solid Ne were recorded. IR fundamentals of trans-ONONO(2) were assigned based on experimental (16/18)O isotopic shifts and guided by DFT calculations. Previously reported contradictory measurements on cis- and trans-ONONO(2) are discussed. Dinitroso peroxide, ONOONO, a proposed intermediate in the IR photoinduced rearrangement of cis-N(2)O(2)O(2) to the various N(2)O(4) species, was not detected. Its absence in the photolysis products indicates a low barrier (0.65 cm(3) g(-1)) is advantageous to efficiently fasten the enzymatic reaction. The targeted specific glycopeptides of the products can be selectively isolated and enriched in B-MOSF through the chemo-affinity between boronic acid and glycol groups, while the non-specific peptides are released to the solutions, or further purified by MOSF and NH(2)-MOSF, which have opposite charges. As a result, the protein digestion and glycol-peptide isolation can be simultaneously achieved in the functionalized macroporous materials in one step, which is a great advantage compared to conventional multi-procedure and time-consuming techniques. PMID- 20024996 TI - Colourimetric carboxylate anion sensors derived from viologen-based receptors. AB - A series of tri- and tetrapodal viologen-based anion receptors showing a colourimetric response to carboxylates, such as acetate, have been synthesised. Alteration of the anion binding sites allows for binding site competition within a receptor. This results in a delayed colourimetric response for urea derivatives compared with pyridinium systems because the anions are initially bound to the periphery of the receptor, away from the viologen unit. DFT calculations and experimental measurements allow the colour change to be assigned to an anion receptor charge-transfer process, facilitated by the exceptionally low reduction potential of the cationic host compounds. Evidence for electron transfer to give the viologen radical cation is also seen in some cases. PMID- 20024997 TI - The sequential building of chiral macrocyclic bis-beta-lactams by double staudinger-Cu-catalyzed azide-alkyne cycloadditions. AB - A novel approach for the synthesis of macrocyclic bis-beta-lactams based on the Cu-catalyzed alkyne-azide cycloaddition (CuAAC) is reported. The procedure is general and allows access to a full range of diastereomerically or enantiomerically pure macrocyclic cavities in good yields. The incorporation of chiral oxazolidinone fragments at C3 in the beta-lactam rings allows the total enantiocontrol of the process. PMID- 20024998 TI - Generation of gas-phase nanosized vanadium oxide clusters from a mononuclear precursor by solution nucleation and electrospray ionization. PMID- 20024999 TI - Reversible dimerization of EGFR revealed by single-molecule fluorescence imaging using quantum dots. AB - The current work explores intermolecular interactions involved in the lateral propagation of cell-signaling by epidermal growth factor receptors (EGFRs). Activation of EGFRs by binding an EGF ligand in the extracellular domain of the EGFR and subsequent dimerization of the EGFR initiates cell-signaling. We investigated interactions between EGFRs in living cells by using single-molecule microscopy, Forster resonance energy transfer (FRET), and atomic force microscopy. By analyzing time-correlated intensity and propagation trajectories of quantum dot (QD)-labeled EGFR single molecules, we found that signaling dimers of EGFR [(EGF-EGFR)(2)] are continuously formed in cell membrane through reversible association of heterodimers [EGF(EGFR)(2)]. Also, we found that the lateral propagation of EGFR activation takes place through transient association of a heterodimer with predimers [(EGFR)(2)]. We varified the transient association between activated EGFR and predimers using FRET from QD-labeled heterodimers to Cy5-labeled predimers and correlated topography and fluorescence imaging. Without extended single-molecule fluorescence imaging and by using bio conjugated QDs, reversible receptor dimerization in the lateral activation of EGFR remained obscured. PMID- 20025000 TI - Dynamic supramolecular polymers based on benzene-1,3,5-tricarboxamides: the influence of amide connectivity on aggregate stability and amplification of chirality. AB - N-Centred benzene-1,3,5-tricarboxamides (N-BTAs) composed of chiral and achiral alkyl substituents were synthesised and their solid-state behaviour and self assembly in dilute alkane solutions were investigated. A combination of differential scanning calorimetry (DSC), polarisation optical microscopy (POM) and X-ray diffraction revealed that the chiral N-BTA derivatives with branched 3,7-dimethyloctanoyl chains were liquid crystalline and the mesophase was assigned as Col(ho). In contrast, N-BTA derivatives with linear tetradecanoyl or octanoyl chains lacked a mesophase and were obtained as crystalline compounds. Variable-temperature infrared spectroscopy showed the presence of threefold, intermolecular hydrogen bonding between neighbouring molecules in the mesophase of the chiral N-BTAs. In the crystalline state at room temperature a more complicated packing between the molecules was observed. Ultraviolet and circular dichroism spectroscopy on dilute solutions of N-BTAs revealed a cooperative self assembly behaviour of the N-BTA molecules into supramolecular polymers with preferred helicity when chiral alkyl chains were present. Both the sergeants-and soldiers as well as the majority-rules principles were operative in stacks of N BTAs. In fact, the self-assembly of N-BTAs resembles closely that of their carbonyl (C=O)-centred counterparts, with the exception that aggregation is weaker and amplification of chirality is less pronounced. The differences in the self-assembly of N- and C=O-BTAs were analysed by density functional theory (DFT) calculations. These reveal a substantially lower interaction energy between the monomeric units in the supramolecular polymers of N-BTAs. The lower interaction energy is due to the higher energy penalty for rotation around the Ph--NH bond compared to the Ph--CO bond and the diminished magnitude of dipole-dipole interactions. Finally, we observed that mixed stacks are formed in dilute solution when mixing N-BTAs and C=O BTAs. PMID- 20025001 TI - Frustrated Lewis pairs: metal-free hydrogen activation and more. AB - Sterically encumbered Lewis acid and Lewis base combinations do not undergo the ubiquitous neutralization reaction to form "classical" Lewis acid/Lewis base adducts. Rather, both the unquenched Lewis acidity and basicity of such sterically "frustrated Lewis pairs (FLPs)" is available to carry out unusual reactions. Typical examples of frustrated Lewis pairs are inter- or intramolecular combinations of bulky phosphines or amines with strongly electrophilic RB(C(6)F(5))(2) components. Many examples of such frustrated Lewis pairs are able to cleave dihydrogen heterolytically. The resulting H(+)/H(-) pairs (stabilized for example, in the form of the respective phosphonium cation/hydridoborate anion salts) serve as active metal-free catalysts for the hydrogenation of, for example, bulky imines, enamines, or enol ethers. Frustrated Lewis pairs also react with alkenes, aldehydes, and a variety of other small molecules, including carbon dioxide, in cooperative three-component reactions, offering new strategies for synthetic chemistry. PMID- 20025002 TI - C(sp(3))-N bond-forming reductive elimination of amines: reactions of bisphosphine-ligated benzylpalladium(II) diarylamido complexes. PMID- 20025003 TI - Highly stable pleated-sheet secondary structure in assemblies of amphiphilic alpha/beta-peptides at the air-water interface. PMID- 20025004 TI - Multistep microchemical synthesis enabled by microfluidic distillation. PMID- 20025005 TI - A polysaccharide-based container transportation system powered by molecular motors. PMID- 20025006 TI - A toxic disinfection by-product, 2,6-dichloro-1,4-benzoquinone, identified in drinking water. PMID- 20025007 TI - A three-coordinate copper(II) amide from reductive cleavage of a nitrosamine. PMID- 20025008 TI - Contact electrification between identical materials. PMID- 20025009 TI - Genetically encoded alkenes in yeast. PMID- 20025010 TI - Mechanistic insights into the formation of InP quantum dots. PMID- 20025011 TI - Room temperature C-H activation and cross-coupling of aryl ureas in water. PMID- 20025012 TI - Isolation of a small carbon nanotube: the surprising appearance of D(5h)(1) C(90). PMID- 20025013 TI - N-Acyliminium ion chemistry: highly efficient and versatile carbon-carbon bond formation by nucleophilic substitution of hydroxy groups catalyzed by Sn(NTf(2))(4). PMID- 20025014 TI - Pharmacological and genetic analysis of mechanisms underlying detrusor overactivity in rats. AB - AIMS: Suprapontine lesions, such as those resulting from cerebrovascular disease, cause bladder storage dysfunction. Detrusor overactivity (DO) following cerebral infarction may be explained by impairment of the suprapontine regulatory system. However, precise mechanisms underlying DO is not clear. The following studies were undertaken to examine pharmacological and genetic mechanisms of DO in rats. RESULTS AND CONCLUSIONS: Mechanisms of long-lasting DO in rats with cerebral infarction require signal transfer, which begins with the opening of glutamate receptors at the dorsal pontine tegmentum. DO induced by cerebral infarction has been proven to be accompanied by an increase in c-fos and zif268 expression in the dorsal pontine tegmentum and periaqueductal gray and mediated by the activation of N-methyl-D-aspartate (NMDA) receptors, cyclooxygenase-2 (COX-2), and prostaglandin E synthase (PGES). Therefore, the arachidonic acid cascade is dynamically activated in the brain after brain ischemia. Bladder sensory pathways are potential targets for drugs used to treat various bladder dysfunctions because of their role in storage symptoms (i.e., urgency, frequency) and in triggering reflex bladder activity. Antimuscarinic drugs and alpha(1)-blockers are the main treatments for overactive bladder, a condition caused by neurologic lesions, aging, bladder outlet obstruction, and other pathologies. These drugs affect sensory bladder storage symptoms, suggesting an action on bladder and urethral afferent pathways. Using animal models of DO, we demonstrated that these drugs improved bladder storage function via suppression of C-fiber afferent nerves from the lower urinary tract. PMID- 20025016 TI - CONSORTing with a QUOROM of MOOSES: the standards movement in scientific reporting. PMID- 20025017 TI - Pelvic organ prolapse and overactive bladder. AB - AIMS: In this review we try to shed light on the following questions: *How frequently are symptoms of overactive bladder (OAB) and is detrusor overactivity (DO) present in patients with pelvic organ prolapse (POP) and is there a difference from women without POP? *Does the presence of OAB symptoms depend on the prolapsed compartment and/or stage of the prolapse? *What is the possible pathophysiology of OAB in POP? *Do OAB symptoms and DO change after conservative or surgical treatment of POP? METHODS: We searched on Medline and Embase for relevant studies. We only included studies in which actual data about OAB symptoms were available. All data for prolapse surgery were without the results of concomitant stress urinary incontinence (SUI) surgery. RESULTS: Community- and hospital-based studies showed that the prevalence of OAB symptoms was greater in patients with POP than without POP. No evidence was found for a relationship between the compartment or stage of the prolapse and the presence of OAB symptoms. All treatments for POP (surgery, pessaries) resulted in an improvement in OAB symptoms. It is unclear what predicts whether OAB symptoms disappear or not. When there is concomitant DO and POP, following POP surgery DO disappear in a proportion of the patients. Bladder outlet obstruction is likely to be the most important mechanism by which POP induces OAB symptoms and DO signs. However, several other mechanisms might also play a role. CONCLUSIONS: There are strong indications that there is a causal relationship between OAB and POP. PMID- 20025018 TI - What was "hot" at the ICS Annual Meeting San Francisco 2009. PMID- 20025015 TI - Beyond neurons: Involvement of urothelial and glial cells in bladder function. AB - AIM: The urothelium, or epithelial lining of the lower urinary tract (LUT), is likely to play an important role in bladder function by actively communicating with bladder nerves, smooth muscle, and cells of the immune and inflammatory systems. Recent evidence supports the importance of non-neuronal cells that may extend to both the peripheral and central processes of the neurons that transmit normal and nociceptive signals from the urinary bladder. Using cats diagnosed with a naturally occurring syndrome termed feline interstitial cystitis (FIC), we investigated whether changes in physiologic parameters occur within 3 cell types associated with sensory transduction in the urinary bladder: 1) the urothelium, 2) identified bladder dorsal root ganglion (DRG) neurons and 3) grey matter astrocytes in the lumbosacral (S1) spinal cord. As estrogen fluctuations may modulate the severity of many chronic pelvic pain syndromes, we also examined whether 17beta-estradiol (E2) alters cell signaling in rat urothelial cells. RESULTS: We have identified an increase in nerve growth factor (NGF) and substance P (SP) in urothelium from FIC cats over that seen in urothelium from unaffected (control) bladders. The elevated NGF expression by FIC urothelium is a possible cause for the increased cell body size of DRG neurons from cats with FIC, reported in this study. At the level of the spinal cord, astrocytic GFAP immuno-intensity was significantly elevated and there was evidence for co expression of the primitive intermediate filament, nestin (both indicative of a reactive state) in regions of the FIC S1 cord (superficial and deep dorsal horn, central canal and laminae V-VIl) that receive input from pelvic afferents. Finally, we find that E2 triggers an estrus-modifiable activation of p38 MAPK in rat urothelial cells. There were cyclic variations with E2-mediated elevation of p38 MAPK at both diestrus and estrus, and inhibition of p38 MAPK in proestrous urothelial cells. CONCLUSION: Though urothelial cells are often viewed as bystanders in the processing of visceral sensation, these and other findings support the view that these cells function as primary transducers of some physical and chemical stimuli. In addition, the pronounced activation of spinal cord astrocytes in an animal model for bladder pain syndrome (BPS) may play an important role in the pain syndrome and open up new potential approaches for drug intervention. PMID- 20025019 TI - Compendium of review articles and the new standardisation report conducted between the International Urogynaecological Association (IUGA) and the International Continence Society (ICS) dealing with the new suggested terminology for female pelvic floor dysfunction. PMID- 20025020 TI - Fourth International Consultation on Incontinence Recommendations of the International Scientific Committee: Evaluation and treatment of urinary incontinence, pelvic organ prolapse, and fecal incontinence. PMID- 20025021 TI - Neurologic urinary incontinence. AB - INTRODUCTION: This manuscript summarizes the work of Committee 10 on neurologic bladder and bowel of the International Consultation on Incontinence in 2008-2009. As the data are very large the outcome is presented in different manuscripts. This manuscript deals with neurologic urinary incontinence. METHODS: Through in debt literature review all aspects of neurological urinary incontinence were studied for levels of evidence. Recommendations for diagnosis and treatment, and for future research were made. RESULTS: Pathophysiology was summarized for different levels of lesions. For epidemiology, specific diagnostics, conservative treatment and surgical treatment of neurologic urinary incontinence, levels of evidence and grades of recommendation were made following ICUD criteria. CONCLUSIONS: Though data are available that advice and guide in the management of urinary incontinence in neurologic patients, not many data have a high level of evidence or permit a high grade of recommendation. More and well-structured research is needed. PMID- 20025022 TI - Neurologic fecal incontinence. AB - INTRODUCTION: This manuscript summarizes the work of Committee 10 on neurologic bladder and bowel of the International Consultation on Incontinence in 2008-2009. As the data are very large the outcome is presented in different manuscripts. This manuscript deals with neurologic fecal incontinence (FI). METHODS: Through in debt literature review all aspects of neurologic urinary and FI were studied for levels of evidence. Recommendations for diagnosis and treatment, and for future research were made. RESULTS: Pathophysiology was summarized for different levels of lesions. For epidemiology, specific diagnostics, conservative treatment, and surgical treatment of neurologic FI levels of evidence and grades of recommendation were made. CONCLUSIONS: Though data are available that advice and guide in the management of FI in neurologic patients, not many data are with a high level of evidence or high grade of recommendation. More and well structured research is needed. PMID- 20025023 TI - Interstitial cells in the urinary bladder--localization and function. AB - AIMS: This review summarizes the currently available literature on the localization and proposed functions of a novel group of cells in the urinary bladder known as interstitial cells or interstitial cells of Cajal (ICC). METHODS: On-line searches of "Pubmed" for bladder, c-Kit, ICC, interstitial cell and myofibroblast were performed to identify relevant studies for the review. RESULTS: The literature contains substantial data that several sub-populations of ICC are present in the wall of the mammalian urinary bladder. These are located in the lamina propria and within the detrusor with distinctive cell shapes and morphological arrangements. Bladder ICC are identified with transmission electron microscopy or by immunohistochemical labeling using antibodies to the Kit receptor which is an established ICC marker. Lamina propria-ICC form a loose network connected via Cx43 gap junctions and are associated with mucosal nerves. Detrusor ICC track the smooth muscle bundles and make frequent contacts with intramural nerves. Both groups of ICC exhibit spontaneous electrical and Ca2+ signalling and also respond to application of neurotransmitter substances including ATP and carbachol. There is emerging evidence that the expression of ICC is upregulated in pathophysiological conditions including the overactive bladder. CONCLUSIONS: There is now a convincing body of evidence that specialized ICC are present in the urinary bladder making important associations with other cells that make up the bladder wall and possessing physiological properties consistent with a role of bladder activity modulation. PMID- 20025025 TI - Neural control of the lower urinary and gastrointestinal tracts: supraspinal CNS mechanisms. AB - Normal urinary function is contingent upon a complex hierarchy of CNS regulation. Lower urinary tract afferents synapse in the dorsal horn of the spinal cord and ascend to the midbrain periaqueductal gray (PAG), with a separate nociception path to the thalamus. A spino-thalamo-cortical sensory pathway is present in some primates, including humans. In the brainstem, the pontine micturition center (PMC) is a convergence point of multiple influences, representing a co-ordinating center for voiding. Many PMC neurones have characteristics necessary to categorize the center as a pre-motor micturition nucleus. In the lateral pontine brainstem, a separate region has some characteristics to suggest a "continence center." Cerebral control determines that voiding is permitted if necessary, socially acceptable and in a safe setting. The frontal cortex is crucial for decision making in an emotional and social context. The anterior cingulate gyrus and insula co-ordinate processes of autonomic arousal and visceral sensation. The influence of these centers on the PMC is primarily mediated via the PAG, which also integrates bladder sensory information, thereby moderating voiding and storage of urine, and the transition between the two phases. The parabrachial nucleus in the pons is also important in behavioral motivation of waste evacuation. Lower urinary tract afferents can be modulated at multiple levels by corticolimbic centers, determining the interoception of physiological condition and the consequent emotional motor responses. Alterations in cognitive modulation, descending modulation, and hypervigilance are important in functional (symptom-based) clinical disorders. PMID- 20025026 TI - Surgical treatment of stress incontinence in men. AB - AIMS: The committee was charged with the responsibility of reviewing and evaluating all published data relating to surgical treatment of male urinary incontinence since the previous consultation in 2004. METHODS: Articles from peer reviewed journals, abstracts from scientific meetings, and literature searches by hand and electronically formed the basis of this review. The articles were evaluated using Levels of Evidences adapted by the ICUD from the Oxford Centre for Evidence Based Medicine. The Recommendations for Care were based on the level of evidence and discussed among the committee members to reach consensus. The incontinence problems were classified according to their etiology, that is, either primarily sphincter or bladder related. RESULTS: Specialist evaluation of the patient is primarily a clinical approach with history, frequency-volume chart, physical examination, and post-void residual urine. Other investigations such as radiographic imaging of the lower urinary tract, cystoscopy, and urodynamic studies can provide important information for the clinician. For stress incontinence of various etiologies the artificial urinary sphincter (AUS) has the longest record of satisfactory results. Consideration must be given to the need for revisions for mechanical breakdown, erosion/infection, and recurrent incontinence, as well as cost. Sling procedures are increasingly being reported to have good outcomes for mild to moderate incontinence. Injectable agents have not shown durable results but newer technologies such as volume-adjustable balloons have shown favorable early results. Incontinence following cystectomy with neobladder and pelvic trauma has been treated most commonly with the AUS. CONCLUSIONS: Although the literature is replete with well-done cohort studies, there is a need for prospective randomized clinical trials. Recommendations for trials include standardized workup and outcome measures and complete reporting of adverse events and long-term results. Further research is also needed to elucidate the mechanism of post-prostatectomy incontinence. PMID- 20025024 TI - Neural control of the lower urinary tract: peripheral and spinal mechanisms. AB - This review deals with individual components regulating the neural control of the urinary bladder. This article will focus on factors and processes involved in the two modes of operation of the bladder: storage and elimination. Topics included in this review include: (1) The urothelium and its roles in sensor and transducer functions including interactions with other cell types within the bladder wall ("sensory web"), (2) The location and properties of bladder afferents including factors involved in regulating afferent sensitization, (3) The neural control of the pelvic floor muscle and pharmacology of urethral and anal sphincters (focusing on monoamine pathways), (4) Efferent pathways to the urinary bladder, and (5) Abnormalities in bladder function including mechanisms underlying comorbid disorders associated with bladder pain syndrome and incontinence. PMID- 20025027 TI - Incontinence in the frail elderly: report from the 4th International Consultation on Incontinence. AB - AIMS: To summarize current knowledge on the etiology, assessment, and management of urinary incontinence (UI) in frail older persons. "Frail" here indicates a person with a clinical phenotype combining impaired physical activity, mobility, muscle strength, cognition, nutrition, and endurance, associated with being homebound or in care institutions and a high risk of intercurrent disease, disability, and death. METHODS: Revision of 3rd ICI report using systematic review covering years 2004-2008. RESULTS: We focus on the etiologic, management, and treatment implications of the key concept that UI in frail persons constitutes a syndromic model with multiple interacting risk factors, including age-related physiologic changes, comorbidity, and the common pathways between them. We present new findings with evidence summaries based on all previous data, and an evidence-based algorithm for evaluation and management of UI in frail elderly. We also present new evidence and summarize the data on etiology and management of nocturia and urinary retention in frail elderly. CONCLUSIONS: Despite the overwhelming burden of UI among this population, there remains a continuing paucity of new clinical trials in frail elderly, limiting evidence for the effectiveness of the full range of UI therapy. Future research is needed on current UI treatments (especially models of care delivery, and pharmacologic and surgical therapies), novel management approaches, and the etiologic mechanisms and pathways of the syndromic model. PMID- 20025028 TI - International Consultation on Urological Diseases: Evidence-based medicine overview of the main steps for developing and grading guideline recommendations. PMID- 20025029 TI - Bladder Pain Syndrome Committee of the International Consultation on Incontinence. AB - AIMS OF STUDY: The Bladder Pain Syndrome Committee of the International Consultation on Incontinence was assigned the task by the consultation of reviewing the syndrome, formerly known as interstitial cystitis, in a comprehensive fashion. This included the topics of definition, nomenclature, taxonomy, epidemiology, etiology, pathology, diagnosis, symptom scales, outcome assessment, principles of management, specific therapies, and future directions in research. STUDY DESIGN, MATERIALS, METHODS: The emphasis was on new information developed since the last consultation 4 years previously. Where possible, existing evidence was assessed and a level of recommendation was developed according to the Oxford system of classification. RESULTS: The consultation decided to refer to the condition as "bladder pain syndrome" (BPS) because the designation is more descriptive of the clinical condition and better fits standard classification taxonomy. Reasonable definitions of BPS include the definition of the ESSIC European group and a slight modification made at a SUFU sponsored Miami meeting in early 2008. Males or females with pain, pressure, or discomfort that they perceive to be related to the bladder with at least one urinary symptom, such as frequency not obviously related to high fluid intake, or a persistent urge to void should be evaluated for possible BPS. The initial assessment consists of a frequency/volume chart, focused physical examination, urinalysis, and urine culture. Urine cytology and cystoscopy are recommended if clinically indicated. Treatment progresses from conservative management through various oral and intravesical therapies, with most surgical therapies reserved for unresponsive cases. Pain management is critical throughout the treatment process. The consultation believes that the disorder is best viewed as one of a group of chronic pain syndromes, rather than as primarily an inflammatory bladder disorder. Recommendations for future research pathways are suggested. PMID- 20025030 TI - Risk of tape-related complications after TVT is at least 4%. PMID- 20025031 TI - Management of fecal incontinence in adults. AB - This article summarises the findings from the Conservative Management of Faecal Incontinence in Adults Committee of the International Consultation on Incontinence. We conducted comprehensive literature searches using the following keywords combined with the relevant intervention: "anal, anorectal, bowel, faecal, fecal, rectal, stool" and "continent$ or incontinent$," Prevalence etimates for faecal or anal incontinence vary widely, from 2.2% to 2.5%. Expert opinion supports the use of general health education, patient teaching about bowel function and advice on lifestyle modification, but the evidence base is small. Unlike urinary incontinence, few "lifestyle" associations have been identified with FI and little is known about whether interventions designed to reduce potential risk factors might improve FI. The article summarises the evidence and recommendations from the committee for clinical practice and future research. PMID- 20025032 TI - Cross-talk and sensitization of bladder afferent nerves. AB - The coordination of pelvic physiologic function requires complex integrative sensory pathways that may converge both peripherally and/or centrally. Following a focal, acute irritative or infectious pelvic insult, these same afferent pathways may produce generalized pelvic sensitization or cross-sensitization as we show bi-directionally for the bladder and bowel in an animal model. Single unit bladder afferent recordings following intracolonic irritation reveal direct sensitization to both chemical and mechanical stimuli that's dependent upon both intact bladder sensory (C-fiber) innervation and neuropeptide content. Concurrent mastocytosis (preponderantly neurogenic) likely plays a role in long-term pelvic organ sensitization via the release of nociceptive and afferent-modulating molecules. Prolonged pelvic sensitization as mediated by these convergent and antidromic reflexive pathway may likewise lead to chronic pelvic pain and thus the overlap of chronic pelvic pain disorders. PMID- 20025034 TI - Investigating afferent nerve activity from the lower urinary tract: highlighting some basic research techniques and clinical evaluation methods. AB - AIMS: To give a review of some basic research recording and clinical evaluations of bladder afferent nerves and the sensory information related to them. METHODS: Literature survey. RESULTS: Direct investigation of the afferent nerve pathways of the lower urinary tract (LUT) can be done in animal studies where potentials can be recorded and measured directly in the dorsal roots after laminectomy. Differentiation between A delta and C fibers is possible when conduction speed is determined. In humans afferent innervation can be studied clinically with determination of the sensation on sensation-related bladder diary, during cystometrical bladder filling, with local electrical stimulation. All need further study. Electrodiagnostic tests are further explored. CONCLUSIONS: Both basic research and clinical evaluation of afferent nerves and sensory function in the LUT are possible. To find out how both relate to each other, and how this function can be evaluated, is the task to be done now. PMID- 20025035 TI - Detrusor myocyte activity and afferent signaling. AB - AIMS: To discuss (1) mechanisms involved in the generation and control of myocyte contractions and consequent afferent nerve activity and (2) these mechanisms as targets for drugs aimed for treatment of overactive bladder (OAB) symptoms and detrusor overactivity (DO). METHODS: Literature review of myocyte activation, bladder afferent nerves, mediators in the bladder, and translational aspects of the findings. RESULTS: During bladder filling, there is normally no parasympathetic outflow from the spinal cord. Despite this, the bladder develops tone during filling and also exhibits non-synchronized local contractions and relaxations that are caused by a basal myogenic mechanical activity that may be reinforced by release of, for example, acetylcholine from non-neuronal and/or neuronal sources or local mediators, such as prostaglandins and endothelins. It is suggested that these spontaneous contractions are able to generate activity in afferent nerves ("afferent noise") that may contribute to DO and OAB. CONCLUSIONS: Spontaneous bladder myocyte contractions and factors that are able to modulate them, as well as the consequent afferent nerve activity, may be targets for drugs meant for treatment of OAB/DO. PMID- 20025033 TI - Changes in afferent activity after spinal cord injury. AB - AIMS: To summarize the changes that occur in the properties of bladder afferent neurons following spinal cord injury. METHODS: Literature review of anatomical, immunohistochemical, and pharmacologic studies of normal and dysfunctional bladder afferent pathways. RESULTS: Studies in animals indicate that the micturition reflex is mediated by a spinobulbospinal pathway passing through coordination centers (periaqueductal gray and pontine micturition center) located in the rostral brain stem. This reflex pathway, which is activated by small myelinated (Adelta) bladder afferent nerves, is in turn modulated by higher centers in the cerebral cortex involved in the voluntary control of micturition. Spinal cord injury at cervical or thoracic levels disrupts voluntary voiding, as well as the normal reflex pathways that coordinate bladder and sphincter function. Following spinal cord injury, the bladder is initially areflexic but then becomes hyperreflexic due to the emergence of a spinal micturition reflex pathway. The recovery of bladder function after spinal cord injury is dependent in part on the plasticity of bladder afferent pathways and the unmasking of reflexes triggered by unmyelinated, capsaicin-sensitive, C-fiber bladder afferent neurons. Plasticity is associated with morphologic, chemical, and electrical changes in bladder afferent neurons and appears to be mediated in part by neurotrophic factors released in the spinal cord and the peripheral target organs. CONCLUSIONS: Spinal cord injury at sites remote from the lumbosacral spinal cord can indirectly influence properties of bladder afferent neurons by altering the function and chemical environment in the bladder or the spinal cord. PMID- 20025036 TI - The emotional motor system and micturition control. AB - Micturition is, similar to all other movements of the body, the result of activation of the motor system in the central nervous system. This review explains how the brain and brainstem control micturition. The basic reflex system begins with a distinct cell group called Gert's Nucleus (GN) in the sacral cord. GN receives information about bladder contents via A-delta fibers from the bladder and bladder sphincter and relays this information to the central part of the midbrain periaqueductal gray (PAG), but not to the thalamus. The PAG, in turn, in case of substantial bladder filling, excites the pontine micturition center (PMC), which cell group, via its long descending pathways to the sacral cord, induces micturition. Higher brain regions in prefrontal cortex and limbic system, by means of its projections to the PAG are able to interrupt this basic reflex system. It allows the individual to postpone micturition until time and place are appropriate. Lesions in the pathways from prefrontal cortex and limbic system to the PAG probably cause urge-incontinence in the elderly. PMID- 20025037 TI - Introduction to special issue on developmental and reproductive toxicity study designs for pharmaceuticals. PMID- 20025038 TI - Embryo-fetal developmental toxicity study design for pharmaceuticals. AB - Assessment of potential developmental and reproductive toxicity of human pharmaceuticals is currently guided by the International Conference on Harmonization (ICH) S5(R2) document (available at http://www.ich.org). The studies that assess developmental hazard are generally conducted in rodents and rabbits. Based on the authors' collective experience, adequate designs (including range-finding studies) and the presentation of data for these studies are described in detail. In addition, the suggested initiation and then total duration of these studies in relation to clinical studies that enroll women of childbearing potential are described. Optional parameters that may be included in the studies are discussed, as are study designs that combine assessments of fertility and developmental toxicity. New methods that may replace or enhance current procedures are outlined. The details described herein will assist all laboratories performing these studies, individuals who need to plan for the studies, and regulatory agencies that ultimately review these studies. PMID- 20025039 TI - The nonclinical fertility study design for pharmaceuticals. AB - Assessment of potential developmental and reproductive toxicity of human pharmaceuticals is currently guided by the ICH S5(R2) document, "Detection of Toxicity to Reproduction for Medicinal Products and Toxicity to Male Fertility." Studies that assess a candidate drug's effect on fertility are generally conducted in rats. The evolution of, and ultimate harmonization of, fertility study designs are reviewed, and specific elements of an acceptable design, as well as the recommendations for presentation of data, are described in detail. Additionally, the timing of nonclinical fertility studies in relation to clinical studies that enroll men and women of reproductive potential is reviewed. Possible strategies for combining fertility assessment with other study designs are also presented. This article provides testing laboratories, sponsors, and regulatory agencies with a comparison of current methods and designs, with the aim of providing a common understanding of the critical design features. PMID- 20025040 TI - Pre- and postnatal developmental toxicity study design for pharmaceuticals. AB - Assessment of potential developmental and reproductive toxicity of human pharmaceuticals is currently guided by the ICH S5(R2) document. The studies that assess the hazard of both pre- and postnatal exposure are predominantly conducted in rodents (rat and mouse). Utilizing the collective experience of the authors, acceptable designs for both the range-finding and definitive studies are presented with detailed descriptions for the presentation of data. In addition, the suggested initiation and then total duration of these studies in relation to clinical studies are described. Optional parameters that may be included in the studies, as well as possible combination with other study designs are discussed. The details described herein will assist all laboratories performing these studies, individuals who need to plan for the studies, and regulatory agencies that ultimately review these studies. PMID- 20025041 TI - Translational systems biology and voice pathophysiology. AB - OBJECTIVES/HYPOTHESIS: Personalized medicine has been called upon to tailor healthcare to an individual's needs. Evidence-based medicine (EBM) has advocated using randomized clinical trials with large populations to evaluate treatment effects. However, due to large variations across patients, the results are likely not to apply to an individual patient. We suggest that a complementary, systems biology approach using computational modeling may help tackle biological complexity in order to improve ultimate patient care. The purpose of the article is: 1) to review the pros and cons of EBM, and 2) to discuss the alternative systems biology method and present its utility in clinical voice research. STUDY DESIGN: Tutorial. METHODS: Literature review and discussion. RESULTS: We propose that translational systems biology can address many of the limitations of EBM pertinent to voice and other health care domains, and thus complement current health research models. In particular, recent work using mathematical modeling suggests that systems biology has the ability to quantify the highly complex biologic processes underlying voice pathophysiology. Recent data support the premise that this approach can be applied specifically in the case of phonotrauma and surgically induced vocal fold trauma, and may have particular power to address personalized medicine. CONCLUSIONS: We propose that evidence around vocal health and disease be expanded beyond a population-based method to consider more fully issues of complexity and systems interactions, especially in implementing personalized medicine in voice care and beyond. PMID- 20025042 TI - Characterization of human papillomavirus type 11-specific immune responses in a preclinical model. AB - OBJECTIVES/HYPOTHESIS: Human papillomavirus (HPV) types 6 and 11 are associated with recurrent respiratory papillomatosis (RRP). Although a prophylactic vaccine has been developed that protects against HPV infection, a therapeutic vaccine is still needed for those patients infected with and/or suffering from persistent disease. Therefore, we developed a novel, therapeutic DNA vaccine targeting HPV 11 and characterized the in vivo immunologic responses generated against HPV-11 E6 and E7 after DNA vaccination in a preclinical model. METHODS: We generated a DNA vaccine that encodes the HPV-11 E6 and E7 genes in a pcDNA3 backbone plasmid. We then vaccinated C57BL/6 mice with the pcDNA3-HPV11-E6E7 DNA plasmid. Splenocytes were harvested from these vaccinated animals and were incubated with overlapping peptides spanning either the HPV-11 E6 or E7 protein. The frequency of interferon-gamma-releasing CD8(+) T cell responses was then analyzed by flow cytometry. RESULTS: Vaccinated mice with the HPV11-E6E7 DNA generated strong CD8(+) T cell responses against the E6(aa44-51) peptide. We determined that the epitope is presented by the MHC class I H2-K(b) molecule. No significant E7 peptide-specific T cell responses were observed. CONCLUSIONS: We developed a novel DNA vaccine that targets the E6 gene of HPV-11. Characterization of the immunologic responses elicited by this DNA vaccine reveals that the E6(aa44-51) peptide contains the most immunogenic region for the HPV-11 viral type. Knowledge of this specific T cell epitope and generation of a RRP preclinical model will allow for the development and evaluation of novel vaccine strategies targeting the RRP patient population. PMID- 20025043 TI - Reconstruction of significant saddle nose deformity using autogenous costal cartilage graft with incorporated mirror image spreader grafts. PMID- 20025044 TI - Insular thyroid carcinoma in a patient with Cowden syndrome. AB - Cowden syndrome is an autosomal dominant disorder characterized by benign and malignant hamartomatous lesions that can develop from all three germ cell derivatives. This disorder predisposes patients to develop malignant tumors of the breast, endometrium, and thyroid. We present a patient with clinically relevant manifestations of Cowden syndrome, with genetic verification, impacting by way of airway compromise due to hamartomas, urinary tract abnormalities, and insular thyroid cancer. This case illustrates the value of recognizing Cowden syndrome at an earlier stage when the patient could have received appropriate management to decrease the morbidity of untreated hamartomatous growths, and an elective thyroidectomy would have been a viable option to manage his malignancy. Through this case report, we provide further insight into management of this disorder. PMID- 20025045 TI - Drug-eluting stent fracture: incidence, contributing factors, and clinical implications. AB - Stent fracture has been observed in noncoronary vessels, especially in the superficial femoral and popliteal arteries and with bare metal stents in saphenous vein grafts of coronary arteries. Since the introduction of drug eluting stents, stent fractures have also been reported in small studies and case reports. We reviewed these publications to assess what is known regarding the incidence, contributing factors, and clinical implications of drug-eluting stent fracture in coronary arteries. The reported rate of drug-eluting stent fracture in coronary arteries ranges from 1 to 8%, although much of the available literature is derived from single-center studies that are heterogeneous in their study methods. A higher risk of stent fracture may be associated with the right coronary artery location, excessive tortuosity or angulation of the vessel, overlapping stents, and longer stents. The closed-cell design of the Cypher stent has been associated with increased rigidity that may increase the risk of stent fracture, although these studies did not assess the overall outcomes between the Cypher and Taxus stents in a head-to-head comparison. Stent fracture has been shown by most studies to be associated with a statistically increased incidence of focal in-stent restenosis, and some have shown an increased risk of target lesion revascularization. Other complications observed with stent fracture include stent thrombosis, coronary aneurysms, myocardial infarction, and sudden death. PMID- 20025046 TI - Developmental and reproductive toxicology studies in nonhuman primates. AB - Developmental and reproductive toxicology testing in nonhuman primates (NHPs) has become more common due to the increasing number of biopharmaceuticals in drug development, since NHPs are frequently the only species to express pharmacologic responses similar to humans. NHPs may also be used to help resolve issues associated with small-molecule reproductive toxicology in traditional species (rodents and rabbits). Adequate designs in NHP are presented for developmental toxicity (embryo-fetal development, pre-postnatal development, enhanced pre postnatal development), reproductive toxicity (male and female), and juvenile toxicity studies. Optional parameters that may be included in these studies are discussed, as are new study designs that consolidate multiple aspects of the reproductive assessment and thereby conserve the limited supply of sexually mature NHPs available for testing. The details described will assist scientists in pharmaceutical, regulatory, and contract research organizations who are involved in conducting these unique studies to optimize their design based on case-by-case considerations. PMID- 20025047 TI - Juvenile animal toxicity study designs to support pediatric drug development. AB - The objective of juvenile animal toxicity studies of pharmaceuticals is to obtain safety data, including information on the potential for adverse effects on postnatal growth and development. Studies in juvenile animals may assist in identifying postnatal developmental toxicities or other adverse effects that are not adequately assessed in the routine toxicity evaluations and that cannot be safely or adequately measured in pediatric clinical trials. Unlike the traditional reproductive and developmental toxicology studies that have been discussed in the accompanying reports, the design requirements for toxicity studies in juvenile animals are not explicitly defined in regulatory guidance. However, studies in juvenile animals can be useful in providing safety information necessary to enable pediatric clinical trials in pediatric patients or when there are special concerns for toxicities that cannot be safely or adequately measured in clinical trials. These juvenile animal toxicity studies are designed on a case-by-case basis. General design considerations and examples of study designs for assessment of juvenile animal toxicity are discussed. PMID- 20025048 TI - Sea urchin embryos exposed to thalidomide during early cleavage exhibit abnormal morphogenesis later in development. AB - BACKGROUND: Clinical use of thalidomide has increased drastically, pushing the questions concerning the teratogenic mechanisms of this drug back to the forefront. Progress in understanding the teratogenic mechanisms has been slow, with the lack of non-primate vertebrate animal models susceptible to the classic reduction deformities remaining a concern. Sea urchin embryos have been used as model organisms for developmental studies for the last century. Like vertebrates, they are deuterostomes and share similar developmental and signaling pathways suggesting they may be an effective system for thalidomide studies. Therefore, we tested sea urchin embryos to see if they were sensitive to the effects of thalidomide. METHODS: Sea urchin embryos were obtained using standard spawning and fertilization techniques. Thalidomide dissolved in DMSO was added to embryo cultures either at fertilization or during early cleavage. Samples of the embryos were evaluated during specific development stages. RESULTS: Lytechinus pictus embryos exposed to 400 microM thalidomide at fertilization or within a window during early cleavage (2-6 hours post-fertilization) exhibit significant levels of abnormal embryos (60-82%) at the pluteus stage, compared to controls levels (< or =10%). Strongylocentrotus purpuratus embryos exposed at initial fertilization or during early cleavage (2-6 hours post-fertilization) exhibit similar responses with significant abnormal levels ranging from (55-70%) at pluteus stage. CONCLUSIONS: Both species of sea urchin tested were susceptible to thalidomide induced teratogenesis during cleavage (4-16 cell stages). This response during cleavage stages warrants further study and indicates that sea urchin embryos may prove to be a useful tool for studying thalidomide effects early in development. PMID- 20025049 TI - Pseudomonas aeruginosa serological analysis in young children with cystic fibrosis diagnosed through newborn screening. AB - BACKGROUND: With newborn screening (NBS) for cystic fibrosis (CF), eradication of Pseudomonas aeruginosa (PA) is possible if PA detection occurs early. A serological response to infection likely precedes culture positivity in CF patients, so PA serological testing is very appealing in this population. However, controversies continue to exist about serology testing, titer cutoffs for enzyme-linked immunosorbent assay (ELISA) antibody tests, and their value in children with CF. METHODS: This longitudinal, prospective study collected respiratory secretions as oropharyngeal swabs or expectorated sputum for culture and also sera over 6 years in 69 patients diagnosed by NBS. Serology assessed PA antibody titers against cell lysate, exotoxin A, and elastase. A novel statistical approach with weighted receiver operating characteristic (ROC) curves was used to determine best antibody titer cutoff values to predict subsequent PA positive cultures. RESULTS: Using these weighted ROC curves, the order of sensitivity was found to be cell lysate, exotoxin A, and then elastase while age specific cutoffs were better than fixed cutoffs previously used. Age-specific serological cutoffs both predict and detect PA respiratory infections with a higher sensitivity and specificity. Serological responses to the PA antigens determined that a response to cell lysate occurs significantly earlier than culture positivity. CONCLUSIONS: Age-specific serological cutoffs rather than fixed values against common PA antigens improve early PA identification in infants and young children diagnosed with NBS. Regular serological assessment with age-specific cutoffs in these children appears to be a worthy diagnostic tool. PMID- 20025050 TI - Reproducibility of multiple breath washout indices in the unsedated preterm neonate. AB - Multiple breath inert gas washout (MBW) is gaining popularity for measurements of resting lung volume and ventilation inhomogeneity. Test reproducibility is an important determinant of the clinical applicability of diagnostic tests. The between-test reproducibility of variables derived from MBW tests in newborn infants is unknown. We aimed to determine the within-test repeatability and short term between-test reproducibility of MBW variables in unsedated preterm infants. We hypothesized that measurements obtained within a 3-day interval in clinically stable preterm infants would be reproducible and suitable for use as an objective clinical outcome measurement. In this cross-sectional observational study, clinically stable hospitalized preterm infants whose parents had given informed consent for MBW studies were tested twice within 72 hr during quiet, unsedated sleep. Functional residual capacity (FRC), lung clearance index (LCI), and the first and second to zeroeth moment ratios (M(1):M(0); M(2):M(0)) were computed from MBW traces obtained using a mainstream ultrasonic flowmeter and 4% sulphur hexafluoride (MBW(SF6)). Within-test repeatability and between-test reproducibility were determined. Within-test repeatability (expressed as a coefficient of variability (C(v))) for differences between two and four replicate measurements on the same test occasion, were 9.3% (FRC), 9.0% (LCI), 7.6% (M(1):M(0)), and 15.6% (M(2):M(0)), respectively. The within-test C(v)'s were not statistically different to the between-tests C(v)'s, which were 7.7% (FRC), 10.3% (LCI), 6.1% (M(1):M(0)), and 13.0% (M(2):M(0)), respectively. Among unsedated preterm infants, between-test reproducibility over a 3-day interval was similar to within-test repeatability. The wide limits of agreement may limit the application of these measures to detect a clinically significant change in condition in small preterm infants. PMID- 20025051 TI - The bronchial circulation--worth a closer look: a review of the relationship between the bronchial vasculature and airway inflammation. AB - Until recently, the bronchial circulation has been relatively ignored in the research and clinical arenas, perhaps because of its small volume and seeming dispensability relative to the pulmonary circulation. Although the bronchial circulation only receives around 1% of the cardiac output in health, it serves functions that are critical to maintaining airway and lung function. The bronchial circulation also plays an important role in many lung and airway diseases; through its ability to increase in size, the bronchial circulation is able to provide lung parenchymal perfusion when the pulmonary circulation is compromised, and more recently the role of the bronchial circulation in the pathogenesis of inflammatory airway disease has been explored. Due to the anatomic variability and small volume of the bronchial circulation, much of the research to date has necessitated the use of animal models and invasive procedures. More recently, non-invasive techniques for measuring bronchial blood flow in the mucosal microvascular network have been developed and offer a new avenue for the study of this circulation in humans. In conjunction with molecular research, measurement of airway blood flow (Q(aw)) may help elucidate the role of the bronchial circulation in inflammatory airway disease and become a useful tool for monitoring therapy. PMID- 20025052 TI - NHLBI training workshop report: The vanishing pediatric pulmonary investigator and recommendations for recovery. AB - The adequacy of the pipeline of advanced pulmonary fellows to supply appropriately trained and committed researchers to enter academic careers was the major topic of a recently held National Heart Lung and Blood Institute NHLBI Workshop: Respiratory Medicine-Related Research Training for Adult and Pediatric Fellows. The special challenges and opportunities for the academic pediatric pulmonary trainee were discussed as part of this workshop and are discussed as a companion paper to the report by the full workshop. Surveys were conducted of pediatric chairs of academic departments and pediatric pulmonary training directors in the United States to examine the current status and opportunities for the pediatric pulmonary trainee. Strategies for recruitment and retention of talented young trainees and junior faculty are proposed. PMID- 20025053 TI - A reproducible technique for specific labeling of antigens using preformed fluorescent molecular IgG-F(ab')2 complexes from primary antibodies of the same species. AB - Immunolabeling two different antigens using the indirect approach with antibodies from the same species is not possible as secondary antibodies can bind to either primary target antibodies. In this study, we describe how preformed complexes of primary and secondary labeled antibodies can be used in such circumstances. In this situation, the first antigen is labeled using the conventional indirect method followed by incubation with the preformed primary-secondary antibody complex against the second antigen. To prevent unbound secondary antibody from binding the indirectly-labeled antibodies, resulting in a false positive, we quenched excess secondary antibody with nonimmune murine serum from the species of the primary antibody. Before the formation of the preformed complex, the optimum dilution of both primary and secondary antibodies was determined. Once these concentrations were established, the concentration of nonimmune murine serum required to quench excess unbound secondary was determined. This step was accomplished by first incubating the sample with an antibody against an antigen known to be localized away from the antigen of interest, followed by the preformed complex. If specific staining was seen, other than that expected from the preformed complex, then the concentration of the serum was deemed insufficient for quenching, and increased accordingly. We demonstrate that this approach is successful in determining the optimum conditions for the preformation of ascites and purified monoclonal primary IgG with fluorescently conjugated F(ab')(2). Double immunolabelling of two focal adhesion antigens and two cytoskeletal proteins, with two murine primary antibodies, are presented as examples of the methodology. PMID- 20025054 TI - Infection of the red palm weevil (Rhynchophorus ferrugineus) by the entomopathogenic fungus Beauveria bassiana: a SEM study. AB - The Red Palm Weevil (Rhynchophorus ferrugineus) is a devastating pest of palms in the Mediterranean, Middle East, and Eastern countries. No effective control measures are available. R. ferrugineus has been found naturally infected by the entomopathogenic fungus Beauveria bassiana, but its infection process in this host is unknown. We have studied the infection of R. ferrugineus larvae and adults by B. bassiana using dry conidia and conidia suspensions using scanning electron microscopy (SEM). In early stages, SEM revealed acquisition of B. bassiana conidia by cuticle ornamentation in legs, antennae, and elytra of R. ferrugineus adults. Subsequently, conidia germinated and frequent episodes of hyphal/conidial fusion were found. Appressoria, signs of adhesion and cuticle degradation led to penetration (even direct) and colonization of R. ferrugineus hosts by the fungus. B. bassiana conidiophores were found in a R. ferrugineus cuticle, which indicate the completion of the life cycle of the fungus in the insect host. SEM has proven that dry conidia of B. bassiana is an adequate inoculum for R. ferrugineus infection. SEM revealed that conidia of B. bassiana attached to the cuticle of R. ferrugineus can germinate and differentiate appressoria. PMID- 20025055 TI - The cathepsin K inhibitor AAE581 induces morphological changes in osteoclasts of treated patients. AB - Inhibitors of Cathepsin K (Cat-K) are recognized as an interesting way to inhibit osteoclast (OC) activity. OCs from patients treated with the anticathepsin-K inhibitor AAE581 (balicatib) were found enlarged. They contained numerous vacuoles filled with tartrate resistant acid phosphatase (TRAcP), an intracellular enzyme that terminates the degradation of collagen internalized in OC transcytotic vesicles. In a phase 2 clinical study, 675 patients with postmenopausal osteoporosis received the Cat-K inhibitor AAE581 at 0, 5, 10, 25, or 50 mg/D during 1 year. Eleven patients had a transiliac bone biopsy, studied undecalcified. Histoenzymatic detection of TRAcP was used to identify and count OC number. The histomorphometrist was not aware of the randomization of patients at the time of analysis. OC were unstained in one patient because of a failure in the fixation protocol, but easily observable in the 10 remaining patients. Whatever the received dose, treated patients exhibited a characteristic aspect of the OC cytoplasm which appeared filled of deeply-stained brown vacuoles, making cells looking like bunches of grape. These round vacuoles, evidenced on TRAcP stained sections, were due to the accumulation of intracytoplasmic TRAcP. This led to a moderate enlargement of the OC size when compared to a series of control osteoporotic patients. AAE581 did not induce OC apoptosis at any dosage but it modified OC morphology. Cat-K inhibition (inhibiting the extracellular collagen breakdown) is associated with a compensatory accumulation of intracellular TRAcP that could not be used to complete protein degradation. TRAcP is also known to be degraded by Cat-K. PMID- 20025056 TI - Curcumin protects against hepatic and renal injuries mediated by inducible nitric oxide synthase during selenium-induced toxicity in Wistar rats. AB - The aim of this study is to evaluate the effect of curcumin in protecting against selenium-induced toxicity in liver and kidney of Wistar rats. Light microscopy evaluation of selenium alone administered rats showed liver to be infiltrated with mononuclear cells, vacuolation, necrosis, and pronounced degeneration. Control liver sections showed a regular morphology of parenchymal cells with intact hepatocytes and sinusoids. Kidney from selenium alone administered rats showed vacuolar degeneration changes in the epithelial cells, cellular proliferation with fibrosis, thickening of capillary walls, and glomerular tuft atrophy. Such changes were also observed in rats administered with selenium and curcumin simultaneously and rats administered first with selenium and then curcumin 24 h later. Interestingly, such degenerative changes observed in liver and kidney induced by selenium were not seen in rats that were administered with curcumin first and selenium 24 h later. This clearly suggests the protective nature of curcumin against selenium toxicity. To understand the probable mechanism of action of curcumin, we analyzed inducible nitric oxide synthase (iNOS) expression by immunohistochemistry, and the results showed an increased iNOS expression in selenium-alone induced liver and kidney. Such high iNOS levels were inhibited in liver and kidney of rats pretreated with curcumin and then with selenium 24 h later. Based on the histological results, it can be concluded that curcumin functions as a protective agent against selenium-induced toxicity in liver as well as kidney, and this action is probably by the regulatory role of curcumin on iNOS expression. PMID- 20025057 TI - Expression of cyclooxygenase-2 and microsomal prostaglandin-E synthase in amoeboid microglial cells in the developing brain and effects of cyclooxygenase-2 neutralization on BV-2 microglial cells. AB - Microglia express cyclooxygenase-2 (COX-2) and microsomal prostaglandin-E synthase (mPGES-1) but their localization in the amoeboid microglial cells (AMC), considered to be the nascent brain macrophages, in the developing brain has remained unexplored; furthermore, their interrelation and regulation have also remained to be fully elucidated. We show here that AMC in postnatal rat brain constitutively expressed COX-2 and mPGES-1 whose immunoexpression was upregulated in rats given lipopolysaccharide (LPS) injections. Reverse transcriptase polymerase chain reaction and Western blot analysis of the callosal tissue rich in AMC revealed that COX-2 and mPGES-1 mRNA and protein expression was augmented following LPS injections. BV-2 cells also exhibited COX-2 and mPGES-1 expression which was enhanced by LPS. However, in cells treated with LPS coupled with COX-2 neutralization, the mRNA expression levels of COX-2, mPGES-1, tumor necrosis factor-alpha, interleukin-1beta and inducible nitric oxide synthase were significantly suppressed; production of prostaglandin E(2) and reactive oxygen species also decreased. Western blot analysis confirmed the changes of protein levels of the above mediators. Remarkably, COX-2 neutralization concomitantly suppressed the protein expression levels of nuclear factor-kappa B (NF-kappaB), phos-NF-kappaB and phos-IkappaB-alpha as well as translocation of NF-kappaB as determined by flow cytometry. In conclusion, AMC in the developing brain expressed COX-2 and mPGES-1 notably when stimulated by LPS. It is suggested that this may be involved in local inflammation during development. Our results have further shown that COX-2 neutralization may be effective in suppressing production of inflammatory mediators and hence its potential use in alleviating neuroinflammation. PMID- 20025058 TI - Prothrombin kringle-2 induces death of mesencephalic dopaminergic neurons in vivo and in vitro via microglial activation. AB - We have shown that prothrombin kringle-2 (pKr-2), a domain of human prothrombin distinct from thrombin could activate cultured rat brain microglia in vitro. However, little is known whether pKr-2-induced microglial activation could cause neurotoxicity on dopaminergic (DA) neurons in vivo. To address this question, pKr 2 was injected into the rat substantia nigra (SN). Tyrosine hydroxylase (TH) immunohistochemistry experiments demonstrate significant loss of DA neurons seven days after injection of pKr-2. In parallel, pKr-2-activated microglia were detected in the SN with OX-42 and OX-6 immunohistochemistry. Reverse transcription PCR and double-label immunohistochemistry revealed that activated microglia in vivo exhibit early and transient expression of inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2) and several proinflammatory cytokines. The pKr-2-induced loss of SN DA neurons was partially inhibited by the NOS inhibitor N(G)-nitro-L-arginine methyl ester hydrochloride, and the COX-2 inhibitor DuP-697. Extracellular signal-regulated kinase 1/2, c-Jun N-terminal kinase and p38 mitogen-activated protein kinase were activated in the SN as early as 1 hr after pKr-2 injection, and localized within microglia. Inhibition of these kinases led to attenuation of mRNA expression of iNOS, COX-2 and several proinflammatory cytokines, and rescue of DA neurons in the SN. Intriguingly, following treatment with pKr-2 in vitro, neurotoxicity was detected exclusively in co-cultures of mesencephalic neurons and microglia, but not microglia-free neuron-enriched mesencephalic cultures, indicating that microglia are required for pKr-2 neurotoxicity. Our results strongly suggest that microglia activated by endogenous compound(s), such as pKr-2, are implicated in the DA neuronal cell death in the SN. PMID- 20025059 TI - Necroptosis, a novel form of caspase-independent cell death, contributes to neuronal damage in a retinal ischemia-reperfusion injury model. AB - Necroptosis is programmed necrosis triggered by death receptor signaling. We investigated whether necroptosis contributes to neuronal damage and functional impairment in a model of retinal ischemia. METHODS: Sprague-Dawley rats were subjected to raised intra-ocular pressure for 45 min and received intravitreal injections of the specific necroptosis inhibitor, Nec-1, its inactive analogue (Nec-1i) or vehicle. Seven days after ischemia, ERGs were performed and then the eyes were enucleated for histological analysis. In other animals, retinas were subjected to propodium iodide, TUNEL staining or Western Blotting and probed with anti-LC-3 antibody. RESULTS: Retinal ischemia resulted in selective neuronal degeneration of the inner layers. Pretreatment with Nec-1 led to significant preservation in thickness and histoarchitecture of the inner retina and functional improvement compared with vehicle-treated controls. Pretreatment with Nec-1i did not provide histological or functional protection. Post-treatment with Nec-1 also significantly attenuated the ERG b-wave reduction compared with ischemic vehicle controls. Nec-1 had no effect on the number of caspase or TUNEL labelled cells in the ischemic retina but did inhibit the induction of LC-3 II and reduced the number of PI-labelled cells after ischemia. CONCLUSION: Necroptosis is an important mode of neuronal cell death and involves autophagy in a model of retinal ischemia. PMID- 20025060 TI - Endothelial cell heterogeneity of blood-brain barrier gene expression along the cerebral microvasculature. AB - The blood-brain barrier (BBB) refers to the network of microvessels that selectively restricts the passage of substances between the circulation and the central nervous system (CNS). This microvascular network is comprised of arterioles, capillaries and venules, yet the respective contribution of each of these to the BBB awaits clarification. In this regard, it has been postulated that brain microvascular endothelial cells (BMEC) from these different tributaries might exhibit considerable heterogeneity in form and function, with such diversity underlying unique roles in physiological and pathophysiological processes. Means to begin exploring such endothelial differences in situ, free from caveats associated with cell isolation and culturing procedures, are crucial to comprehending the nature and treatment of CNS diseases with vascular involvement. Here, the recently validated approach of immuno-laser capture microdissection (immuno-LCM) coupled to quantitative real-time PCR (qRT-PCR) was used to analyze gene expression patterns of BMEC retrieved in situ from either capillaries or venules. From profiling 87 genes known to play a role in BBB function and/or be enriched in isolated brain microvessels, results imply that most BBB properties reside in both segments, but that capillaries preferentially express some genes related to solute transport, while venules tend toward higher expression of an assortment of genes involved in inflammatory-related tasks. Fuller appreciation of such heterogeneity will be critical for efficient therapeutic targeting of the endothelium and the management of CNS disease. PMID- 20025061 TI - Analysis of the expression and function of BRINP family genes during neuronal differentiation in mouse embryonic stem cell-derived neural stem cells. AB - We previously identified a novel family of genes, BRINP1, 2, and 3, that are predominantly and widely expressed in both the central nervous system (CNS) and peripheral nervous system (PNS). In the present study, we analyzed the expression pattern of three BRINP genes during differentiation of mouse embryonic stem (ES) cell-derived neural stem cells (NSCs) and their effects on the cell-cycle regulation of NSCs. While there was no significant expression of any BRINP-mRNA expressed in mouse ES cells, BRINP 1 and 2-mRNAs was expressed at high levels in the ES cell-derived neural stem cells. Upon differentiation into neuronal cells in the presence of retinoic acid and BDNF, all three types of BRINP-mRNA were induced with a similar time course peaking at day three of treatment. Upon differentiation into astroglial cells in the presence of serum, BRINP1-mRNA was slightly up-regulated, while BRINP2- and BRINP3-mRNAs were almost abolished in the astrocytes. While 69.2, 26.1, and 7.7% of cells in a population of NSCs in the exponentially growing phase were in the G1, S and G2 phases, respectively, over-expression of any one of the three BRINP genes completely abolished cells in the G2 phase and significantly reduced the cells in S phase to 11.8-13.8%. Based on these results, the physiological roles of induced BRINP genes in the cell cycle suppression of terminally differentiated post-mitotic neurons are discussed. PMID- 20025063 TI - Role of mast cell activation in inducing microglial cells to release neurotrophin. AB - The brain-derived neurotrophic factor (BDNF) plays a critical role in pain hypersensitivity. BDNF is the ligand of P2X4 receptors (P2X4R) in the microglia. The causative factors involving the P2X4R over expression in the microglia remains unclear. Mast cell activation has a close relation with pain hypersensitivity. However, the underlying mechanism between mast cell activation and pain hypersensitivity is unknown. The present study aimed to elucidate the mechanism by which mast cell activation promoted the expression of P2X4R in the microglia. The results of present study showed that mast cell activation markedly promoted the expression of P2X4R and BDNF in microglial cells, which significantly enhanced the release of BDNF from microglial cells upon exposure to adenosine triphosphate. Mast cell-derived tryptase activated PAR2 that resulted in promoting the expression of P2X4R in microglial cells. Pretreatment with antibodies against tryptase or PAR2, or using tryptase-deficient HMC-1 cells or PAR2-deficient microglial cells abolished the increase in P2X4R expression and BDNF release. Increase in mitogen activated protein kinase phosphorylation was observed in the processes of mast cell-induced BDNF release and P2X4R expression. We conclude that mast cell activation has the capacity to promote the expression of P2X4R and BDNF in microglial cells. PMID- 20025062 TI - High-frequency stimulation of the subthalamic nucleus restores neural and behavioral functions during reaction time task in a rat model of Parkinson's disease. AB - Deep brain stimulation (DBS) has been used in the clinic to treat Parkinson's disease (PD) and other neuropsychiatric disorders. Our previous work has shown that DBS in the subthalamic nucleus (STN) can improve major motor deficits, and induce a variety of neural responses in rats with unilateral dopamine (DA) lesions. In the present study, we examined the effect of STN DBS on reaction time (RT) performance and parallel changes in neural activity in the cortico-basal ganglia regions of partially bilateral DA- lesioned rats. We recorded neural activity with a multiple-channel single-unit electrode system in the primary motor cortex (MI), the STN, and the substantia nigra pars reticulata (SNr) during RT test. RT performance was severely impaired following bilateral injection of 6 OHDA into the dorsolateral part of the striatum. In parallel with such behavioral impairments, the number of responsive neurons to different behavioral events was remarkably decreased after DA lesion. Bilateral STN DBS improved RT performance in 6-OHDA lesioned rats, and restored operational behavior-related neural responses in cortico-basal ganglia regions. These behavioral and electrophysiological effects of DBS lasted nearly an hour after DBS termination. These results demonstrate that a partial DA lesion-induced impairment of RT performance is associated with changes in neural activity in the cortico-basal ganglia circuit. Furthermore, STN DBS can reverse changes in behavior and neural activity caused by partial DA depletion. The observed long-lasting beneficial effect of STN DBS suggests the involvement of the mechanism of neural plasticity in modulating cortico-basal ganglia circuits. PMID- 20025064 TI - What's the best statistic for a simple test of genetic association in a case control study? AB - Genome-wide genetic association studies typically start with univariate statistical tests of each marker. In principle, this single-SNP scanning is statistically straightforward--the testing is done with standard methods (e.g. chi(2) tests, regression) that have been well studied for decades. However, a number of different tests and testing procedures can be used. In a case-control study, one can use a 1 df allele-based test, a 1 or 2 df genotype-based test, or a compound procedure that combines two or more of these statistics. Additionally, most of the tests can be performed with or without covariates included in the model. While there are a number of statistical papers that make power comparisons among subsets of these methods, none has comprehensively tackled the question of which of the methods in common use is best suited to univariate scanning in a genome-wide association study. In this paper, we consider a wide variety of realistic test procedures, and first compare the power of the different procedures to detect a single locus under different genetic models. We then address the question of whether or when it is a good idea to include covariates in the analysis. We conclude that the most commonly used approach to handle covariates--modeling covariate main effects but not interactions--is almost never a good idea. Finally, we consider the performance of the statistics in a genome scan context. PMID- 20025065 TI - Detecting interacting genetic loci with effects on quantitative traits where the nature and order of the interaction are unknown. AB - Standard techniques for single marker quantitative trait mapping perform poorly in detecting complex interacting genetic influences. When a genetic marker interacts with other genetic markers and/or environmental factors to influence a quantitative trait, a sample of individuals will show different effects according to their exposure to other interacting factors. This paper presents a Bayesian mixture model, which effectively models heterogeneous genetic effects apparent at a single marker. We compute approximate Bayes factors which provide an efficient strategy for screening genetic markers (genome-wide) for evidence of a heterogeneous effect on a quantitative trait. We present a simulation study which demonstrates that the approximation is good and provide a real data example which identifies a population-specific genetic effect on gene expression in the HapMap CEU and YRI populations. We advocate the use of the model as a strategy for identifying candidate interacting markers without any knowledge of the nature or order of the interaction. The source of heterogeneity can be modeled as an extension. PMID- 20025066 TI - Influence of persistent thyroxine reduction on spermatogenesis in rats neonatally exposed to 2,2',4,4',5,5'-hexa-chlorobiphenyl. AB - BACKGROUND: The aim of the present study is to determine the long-term testicular effects of neonatal exposure to PCB153. METHODS: Sprague-Dawley (SD) rats were treated by oral gavage with PCB153 in corn oil at doses of 0, 0.025, and 2.5 mg/kg per day from postnatal day 3 (PND 3) to PND7. The rats were sacrificed on PND 8 and PND 77. TUNEL in situ detection for testis apoptosis, immunohistochemical staining of thyroid gland for thyroxine (T4), semi quantitative RT-PCR for mRNA expression, and radioimmunoassay (RIA) for serum hormone levels were performed. RESULTS: Neonatal treatment with PCB153 at both doses had no obvious effects on body weight, testis weight, testis histology, and germ cell apoptosis, but decreased T4 staining in thyroid gland was observed on PND 8. On PND 77, neonatal treatment with 2.5 mg/kg per day of PCB153 significantly reduced daily sperm product (DSP). Serum levels of thyroxine (T4) and free thyroxine (FT4) decreased, but there were no differences in thyroid stimulating hormone (TSH) level between the control and exposed groups. Gap junction connexin43 (CX43) and cyclin-dependent kinase inhibitor (CDKI) P27kip1 mRNA expression, which was associated with Sertoli cell differentiation, was significantly reduced after PCB153 treatment on PND 8 but not on PND 77. Androgen binding protein (ABP) and androgen receptor (AR) mRNA expression, which indicates Sertoli cell maturation, was suppressed on PND 77 after neonatal PCB153 exposure. CONCLUSIONS: The findings in this study suggest that neonatal exposure to PCB153 induces persistent T4 reduction, which disturbs Sertoli cell function, and subsequently results in alterations in adult spermatogenesis. Birth Defects Res (Part B) 89:18-25, 2010. (c) 2009 Wiley-Liss, Inc. PMID- 20025067 TI - Correlations between auditory structures and hearing sensitivity in non-human primates. AB - Primates show distinctions in hearing sensitivity and auditory morphology that generally follow phylogenetic patterns. However, few previous studies have attempted to investigate how differences in primate hearing are directly related to differences in ear morphology. This research helps fill this void by exploring the form-to-function relationships of the auditory system in a phylogenetically broad sample of non-human primates. Numerous structures from the outer, middle, and inner ears were measured in taxa with known hearing capabilities. The structures investigated include the overall size and shape of the pinna, the areas of the tympanic membrane and stapedial footplate, the masses and lever arm lengths of the ossicles, the volumes of the middle ear cavities, and the length of the cochlea. The results demonstrate that a variety of auditory structures show significant correlations with certain aspects of hearing (particularly low frequency sensitivity). Although the majority of these relationships agree with expectations from auditory theory, some traditional (and possibly outdated) ideas were not supported. For example, the common misconception that higher middle ear transformer ratios (e.g., impedance transformer ratio) result in increased hearing sensitivity was not supported. Although simple correlations between form and function do not necessarily imply causality, the relationships defined in this study not only increase our understanding of auditory patterns in extant taxa but also lay the foundation to begin investigating the hearing in fossil primates. PMID- 20025068 TI - Hybrid ultrasound MRI for improved cardiac imaging and real-time respiration control. AB - A hybridized dual-imaging system combining real-time ultrasound imaging and MRI was utilized for cardiac imaging at 1.5 T and 3 T. The ultrasound scanner with a programmable software interface was connected via computer to the MRI scanner. Electronic noise was eliminated with electromagnetic shielding and grounding to the screen room. At 3 T, real-time prospective motion compensation in dynamic cine cardiac imaging was implemented using B-mode ultrasound imaging. The ultrasound technique avoided drawbacks such as signal saturation or steady-state interruption of the MR navigator gating. At 1.5 T, a low-latency real-time feedback to balanced steady state free precision MR imaging was performed in three normal volunteers. Results showed active tracking of the heart during respiratory motion and improvement in time-averaged cardiovascular images. Future studies can fully exploit the potential of the high-frequency position information provided by the ultrasound system for more advanced applications in real-time organ tracking. PMID- 20025069 TI - Guideline for optimization of colorectal cancer surgery and pathology. AB - BACKGROUND AND OBJECTIVES: There is evidence of gaps in care for colorectal cancer surgery related to obtaining negative resection margins and lymph node assessment. Recommendations on the surgical and pathological management of curable colon and rectal cancer were developed. METHODS: A systematic review on colorectal resection margins and lymph nodes was conducted. This evidence, combined with evidence from existing guidelines and expert consensus, was used to develop recommendations. The draft guideline was reviewed by an expert panel and was externally reviewed by practitioners in Ontario, Canada. RESULTS: The search of the recent literature identified 107 articles pertinent to resection margins and lymph node assessment. The majority of the evidence was of poor quality. Of the 63 practitioners who reviewed the guideline, 97% agreed with the draft recommendations and 92% thought that the report should be approved as a practice guideline. CONCLUSIONS: Achieving optimized performance concerning margin status and lymph node assessment requires the coordinated efforts of surgeons and pathologists, as well as other medical professionals. Focus should be on ensuring that colorectal cancers are resected with negative (R0) margins and that an adequate number of lymph nodes are assessed to allow for accurate decision making relating to prognosis and adjuvant therapy. PMID- 20025070 TI - Evidence-based clinical practice guideline development: principles, challenges, and accountability to evidence. PMID- 20025071 TI - Preoperative serum CA-125 levels and risk of suboptimal cytoreduction in ovarian cancer: a meta-analysis. AB - BACKGROUND: This meta-analysis was designed to determine the ability of pretreatment CA-125 level to predict optimal cytoreduction in advanced ovarian cancer (OC). METHODS: Through literature search, 14 studies were identified. In addition, we retrospectively reviewed the data of 154 patients with OC. Using the bi-variate model, diagnostic performance of CA-125 was assessed at the various cut-off levels. An overall odds ratio was obtained using random effects model. RESULTS: A total of 2,192 patients were included in the analysis. The pooled optimal cytoreduction rate and the mean of median CA-125 levels were 53.7% and 580 U/ml, respectively. At the cut-off of 500 U/ml, overall sensitivity and specificity were 68.9% (95% confidence interval [CI] 62.0-75.1%) and 63.2% (95% CI 53.7-71.7%), respectively. Positive and negative likelihood ratios were 1.87 (95% CI 1.40-2.50) and 0.49 (95% CI 0.37-0.66). The CA-125 >500 U/ml showed strong association with a risk of suboptimal cytoreduction with an odds ratio of 3.69 (95% CI 2.02-6.73). CONCLUSIONS: The current analysis indicates that CA-125 is a strong risk factor of suboptimal cytoreduction and it may be applied in preoperative counseling and treatment planning. However, it also shows that CA 125 lacks the ability to predict optimal cytoreduction accurately. PMID- 20025072 TI - Intracisternal A particle genes: Distribution in the mouse genome, active subtypes, and potential roles as species-specific mediators of susceptibility to cancer. AB - Rodents, mice and rats in particular, are the species of choice for evaluating chemical carcinogenesis. However, different species and strains often respond very differently, undermining the logic of extrapolation of animal results to humans and complicating risk assessment. Intracisternal A particles (IAPs), endogenous retroviral sequences, are an important class of transposable elements that induce genomic mutations and cell transformation by disrupting gene expression. Several lines of evidence support a role of IAPs as mouse-specific genetic factors in responses to toxicity and expression of disease phenotypes. Since multiple subtypes and copies of IAPs are present in the mouse genome, their activity and locations relative to functional genes are of critical importance. This study identified the major "active" subtypes of IAPs (subtype 1/1a) that are responsible for newly transposed IAP insertions described in the literature, and confirmed that (1) polymorphisms for IAP insertions exist among different mouse strains and (2) promoter activity of the LTRs can be modulated by chemicals. This study further identified all the genes in the C57BL/6 mouse genome with IAP subtype 1 and 1a sequences inserted in their proximity, and the major biofunctional categories and cellular signaling networks of those genes. Since many "IAP-associated genes" play important roles in the regulation of cell proliferation, cell cycle, and cell death, the associated IAPs, upon activation, can affect cellular responses to xenobiotics and disease processes, especially carcinogenesis. This systemic analysis provides a solid foundation for further investigations of the role of IAPs as species- and strain-specific disease susceptibility factors. PMID- 20025073 TI - Dietary folate and vitamin B6 are not associated with p53 mutations in esophageal adenocarcinoma. AB - Recent studies have suggested an association between dietary folate, and related B-vitamins, and risk for cancer, potentially mediated by the p53 tumor suppressor gene. The aim of this study was to explore the effect of dietary folate and vitamin B(6) intake on p53 in the molecular pathogenesis of esophageal adenocarcinoma (EADC). For each participant, a structured questionnaire was used to obtain detailed sociodemographic and lifestyle risk factors, including diet, from which folate and vitamin B(6) intake were calculated. Risks for p53 mutations, p53 mutations at CpG sites, and p53 protein overexpression among EADC cases (n = 54) were calculated using logistic regression with dietary folate and vitamin B(6) intake as predictive variables, adjusting for age, gender, smoking, and alcohol consumption. No significant differences were found for patients with EADC who had p53 mutations (n = 21) compared with patients with wild-type p53 (n = 33) with respect to selected clinicopathologic variables (age, gender, tumor grade, stage, alcohol, or tobacco consumption) and dietary intake of folate or vitamin B(6). No statistically significant associations were seen between dietary folate and vitamin B(6) intake (highest vs. lowest quartiles) and p53 mutations, p53 mutations at CpG sites (n = 12), and p53 protein overexpression (n = 17). We conclude that dietary intake of folate and vitamin B(6) do not appear to have an effect on p53, suggesting alternative molecular mechanisms underlying esophageal adenocarcinogenesis. PMID- 20025074 TI - Airways obstruction among older construction and trade workers at Department of Energy nuclear sites. AB - BACKGROUND: A study of chronic obstructive pulmonary disease (COPD) among 7,579 current and former workers participating in medical screening programs at Department of Energy (DOE) nuclear weapons facilities through September 2008 was undertaken. METHODS: Participants provided a detailed work and exposure history and underwent a respiratory examination that included a respiratory history, respiratory symptoms, a posterior-anterior (P-A) chest radiograph classified by International Labour Office (ILO) criteria, and spirometry. Statistical models were developed to generate group-level exposure estimates that were used in multivariate logistic regression analyses to explore the risk of COPD in relation to exposures to asbestos, silica, cement dust, welding, paints, solvents, and dusts/fumes from paint removal. Risk for COPD in the study population was compared to risk for COPD in the general US population as determined in National Health and Nutrition Examination Survey (NHANES III). RESULTS: The age standardized prevalence ratio of COPD among DOE workers compared to all NHANES III data was 1.3. Internal analyses found the odds ratio of COPD to range from 1.6 to 3.1 by trade after adjustment for age, race, sex, smoking, and duration of DOE employment. Statistically significant associations were observed for COPD and exposures to asbestos, silica, welding, cement dusts, and some tasks associated with exposures to paints, solvents, and removal of paints. CONCLUSIONS: Our study of construction workers employed at DOE sites demonstrated increased COPD risk due to occupational exposures and was able to identify specific exposures increasing risk. This study provides additional support for prevention of both smoking and occupational exposures to reduce the burden of COPD among construction workers. PMID- 20025077 TI - Hierarchically constrained dynamics and emergence of complex behavior in nanohybrids. PMID- 20025076 TI - Curcumin inhibits the Sonic Hedgehog signaling pathway and triggers apoptosis in medulloblastoma cells. AB - Medulloblastoma is an aggressive primary brain tumor that arises in the cerebellum of children and young adults. The Sonic Hedgehog (Shh) signaling pathway that plays important roles in the pathology of this aggressive disease is a promising therapeutic target. In the present report we have shown that curcumin has cytotoxic effects on medulloblastoma cells. Curcumin suppressed also cell proliferation and triggered cell-cycle arrest at G(2)/M phase. Moreover, curcumin inhibited the Shh-Gli1 signaling pathway by downregulating the Shh protein and its most important downstream targets GLI1 and PTCH1. Furthermore, curcumin reduced the levels of beta-catenin, the activate/phosphorylated form of Akt and NF-kappaB, which led to downregulating the three common key effectors, namely C myc, N-myc, and Cyclin D1. Consequently, apoptosis was triggered by curcumin through the mitochondrial pathway via downregulation of Bcl-2, a downstream anti apoptotic effector of the Shh signaling. Importantly, the resistant cells that exhibited no decrease in the levels of Shh and Bcl-2, were sensitized to curcumin by the addition of the Shh antagonist, cyclopamine. Furthermore, we have shown that curcumin enhances the killing efficiency of nontoxic doses of cisplatin and gamma-rays. In addition, we present clear evidence that piperine, an enhancer of curcumin bioavailability in humans, potentiates the apoptotic effect of curcumin against medulloblastoma cells. This effect was mediated through strong downregulation of Bcl-2. These results indicate that curcumin, a natural nontoxic compound, represents great promise as Shh-targeted therapy for medulloblastomas. PMID- 20025075 TI - Occupational factors and risk of Parkinson's disease: A population-based case control study. AB - BACKGROUND: Parkinson's disease (PD) has been associated with various workplace factors, but the evidence is inconsistent. OBJECTIVE: To estimate the risk of PD associated with various jobs and workplace exposures. METHODS: We conducted a population-based, case-control study of 404 incident PD cases and 526 age and sex matched controls, collecting self-reported work histories including job titles and exposures to various industrial toxicants. Relative risks of PD from these exposures were estimated with odds ratios (OR) and 95% confidence intervals (CI) using logistic regression. RESULTS: Risk was not significantly affected by farming work, by metal work, or by exposure to pesticides, metals, or solvents. CONCLUSIONS: These findings do not provide support for the hypothesis that workplace factors affect the risk of PD. PMID- 20025078 TI - One-dimensional heterostructures of single-walled carbon nanotubes and CdSe nanowires. PMID- 20025079 TI - Intracellular silicon chips in living cells. PMID- 20025080 TI - Water-dispersible, multifunctional, magnetic, luminescent silica-encapsulated composite nanotubes. AB - A multifunctional one-dimensional nanostructure incorporating both CdSe quantum dots (QDs) and Fe(3)O(4) nanoparticles (NPs) within a SiO(2)-nanotube matrix is successfully synthesized based on the self-assembly of preformed functional NPs, allowing for control over the size and amount of NPs contained within the composite nanostructures. This specific nanostructure is distinctive because both the favorable photoluminescent and magnetic properties of QD and NP building blocks are incorporated and retained within the final silica-based composite, thus rendering it susceptible to both magnetic guidance and optical tracking. Moreover, the resulting hydrophilic nanocomposites are found to easily enter into the interiors of HeLa cells without damage, thereby highlighting their capability not only as fluorescent probes but also as possible drug-delivery vehicles of interest in nanobiotechnology. PMID- 20025081 TI - Ultrathin alumina membranes for surface nanopatterning in fabricating quantum sized nanodots. AB - Using ultrathin alumina membranes (UTAMs) as evaporation or etching masks large scale ordered arrays of surface nanostructures can be synthesized on substrates. However, it is a challenge for this technique to synthesize quantum-sized surface structures. Here an innovative approach to prepare UTAMs with regularly arrayed pores in the quantum size range is reported. This new approach is based on a well controlled pore-opening process and a modulated anodization process. Using UTAMs with quantum-sized pores for the surface patterning process, ordered arrays of quantum dots are synthesized on silicon substrates. This is the first time in realizing large-scale regularly arrayed surface structures in the quantum size range using the UTAM technique, which is an important breakthrough in the field of surface nanopatterning. PMID- 20025082 TI - Missing data in the exposure of interest and marginal structural models: a simulation study based on the Framingham Heart Study. AB - Missing data are common in longitudinal studies and can occur in the exposure interest. There has been little work assessing the impact of missing data in marginal structural models (MSMs), which are used to estimate the effect of an exposure history on an outcome when time-dependent confounding is present. We design a series of simulations based on the Framingham Heart Study data set to investigate the impact of missing data in the primary exposure of interest in a complex, realistic setting. We use a standard application of MSMs to estimate the causal odds ratio of a specific activity history on outcome. We report and discuss the results of four missing data methods, under seven possible missing data structures, including scenarios in which an unmeasured variable predicts missing information. In all missing data structures, we found that a complete case analysis, where all subjects with missing exposure data are removed from the analysis, provided the least bias. An analysis that censored individuals at the first occasion of missing exposure and includes a censorship model as well as a propensity model when creating the inverse probability weights also performed well. The presence of an unmeasured predictor of missing data only slightly increased bias, except in the situation such that the exposure had a large impact on missing data and the unmeasured variable had a large impact on missing data and outcome. A discussion of the results is provided using causal diagrams, showing the usefulness of drawing such diagrams before conducting an analysis. PMID- 20025083 TI - Psychosocial effects of mastectomy on married African women in Northwestern Nigeria. AB - OBJECTIVES: Few studies have examined the psychosocial impact of breast cancer and its treatment on African women who come from a poverty-stricken, uneducated background in a developing country. The purpose of this study was to describe the specific psychosocial effects of breast cancer on married African women in order to help physicians educate and counsel future women and their families in Northwestern Nigeria. METHODS: Two semi-structured, self-report questionnaires were given to 81 consenting married African women treated with unilateral total mastectomy secondary to operable breast cancer at the Ahmadu Bello University Teaching Hospital. Questionnaires were answered at the time of the diagnosis and treatment consultation and at 6 months postmastectomy and included questions about demographics, frequency of conjugal relations and how the woman's sense of femininity was affected. Marital status was also tracked up to 3 years. RESULTS: Six months after surgery, the survey responses revealed that 67.9% of women felt inadequate as a woman because of the mastectomy and that 79.0% experienced a decrease in frequency of conjugal relations. Three years after primary breast cancer treatment, 61.7% of the participants were still married while 38.3% reported being divorced/separated from their husbands. CONCLUSIONS: These results indicated that married African women face significant physical, emotional and social changes and difficulties following primary breast cancer treatment. Culturally sensitive therapeutic groups and interventions should be established to help Nigerian women with breast cancer and their spouses and families understand and cope with the disease and its long-term health and quality-of-life implications. PMID- 20025084 TI - Diethylene glycol in propofol infusion syndrome? PMID- 20025085 TI - [Osteoporosis guidance]. PMID- 20025086 TI - [Prenatal diagnosis and eugenics]. PMID- 20025087 TI - Immunocytochemical methods and protocols. PMID- 20025088 TI - Risperidone and dementia-related aggression: new indication. A last resort, no better than haloperidol. AB - When elderly Alzheimer's patients remain dangerously aggressive despite non-drug measures, there is no evidence that risperidone is safer or any more effective than haloperidol. PMID- 20025089 TI - Lamotrigine and absence seizures: new indication. Try valproic acid first. AB - In a trial including 38 children, lamotrigine, which had to be introduced very gradually in order to reduce the risk of potentially severe skin reactions, took longer than valproic acid to control typical absence seizures. PMID- 20025090 TI - Pantoprazole available without a prescription: new status. Better than H2 receptor agonists, but not for pregnant women. AB - When lifestyle measures and antacids are inadequate, and when no alarm symptoms are present, pantoprazole has a better risk-benefit balance than H2 receptor agonists. PMID- 20025091 TI - Aripiprazole and adolescent schizophrenia: a simple neuroleptic. No comparison with standard neuroleptic in adolescents. PMID- 20025092 TI - Glatiramer: a second look. With longer follow-up: still no proven benefit in multiple sclerosis. PMID- 20025094 TI - Vitamin B6: peripheral neuropathy. Dependent on dose and duration of use. PMID- 20025093 TI - Valproic acid: long-term effects on children exposed in utero. AB - (1) Numerous follow-up studies of pregnancies in women with epilepsy show that valproic acid is more teratogenic than other antiepileptics. The risk of malformations increases with doses above 1000 mg/day; (2) Malformations associated with valproic acid include neural tube defects in 1-2% of exposed children, as well as urogenital, craniofacial and digital abnormalities. Cardiac disorders and limb defects have also been reported; (3) Convergent results of several cohort studies show that exposure to valproic acid in utero has detrimental effects on intelligence, language and behavior, which appear in school-age children; (4) In practice, the use of valproic acid should be avoided throughout pregnancy, as well as by women of childbearing age not using effective contraception. If a woman is planning pregnancy, the choice of valproic acid should be reassessed with the patient. If valproic acid therapy is maintained, the minimum effective daily dose should be determined and folic acid supplementation initiated. PMID- 20025095 TI - Injectable amphotericin B: mix-ups between lipid and non-lipid formulations. AB - (1) Amphotericin B is a standard antifungal drug with a narrow therapeutic margin. It has potentially severe adverse effects, especially renal and cardiac toxicity; (2) Three injectable forms of amphotericin B are currently available in France and elsewhere. They mainly differ in their dose regimens; (3) Cases of fatal overdose have occurred following confusion between the various injectable formulations; (4) Measures intended to prevent such errors include: specific training of healthcare staff; informing users that the different injectable forms of amphotericin B are not interchangeable; simultaneously mentioning the excipient and the international nonproprietary name (INN) and the brand name; remembering the maximum dose of the conventional formulation; noting the dose per kilogram per day, as well as the total daily dose; and if possible, involving the patient. PMID- 20025096 TI - Testosterone gel: children exposed through contact with adults. Virilisation cases were reported. PMID- 20025097 TI - Non-anaemic pregnant women should not take iron supplements. AB - (1) Iron-deficiency anaemia during pregnancy increases the risk of low birth weight and preterm birth; (2) In a randomised double-blind placebo-controlled trial, iron supplementation in pregnant women with haemoglobin levels of at least 13.2 g/100 ml at the beginning of the 2nd trimester was associated with low birth weight and maternal hypertension; (3) In a trial in women with haemoglobin levels of at least 11.5 g/100 ml who took supplemental iron, haemoglobin levels above 14.5 g/100 ml at 28 weeks of gestation were associated with an 8-fold increase in the risk of preterm birth and a 6-fold increase in the risk of low birth weight; (4) An epidemiological study showed a link between high maternal haemoglobin levels and low birth weight; (5) In practice, iron supplements should not be taken by pregnant women whose haemoglobin levels exceed 11 g/100 ml during the 1st and 3rd trimesters and 10.5 g/100 ml during the 2nd trimester. PMID- 20025098 TI - Treatment of multiple myeloma: 2009 update. AB - (1) New marketing authorizations continue to be granted for treatments of multiple myeloma, and new trials and meta-analyses continue to be published. This review re-examines our previous conclusions based on data published between 2003 and 2008. We again used the standard Prescrire methodology to review the latest data; (2) In patients who are symptomatic but who do not qualify for haematopoietic stem cell transplantation (especially people aged over 65), the results of five comparative trials suggest that adding thalidomide to the melphalan-prednisone combination delays myeloma progression by an additional 5 to 10 months. There is possibly also an increase in overall survival time. This triple combination is therefore a first-line treatment option. There is no firm evidence that replacing thalidomide with bortezomib in this combination provides an advantage; (3) In symptomatic patients aged under 65, two meta-analyses have compared high-dose chemotherapy followed by autologous stem cell transplantation with conventional chemotherapy. Unlike early trials, these meta-analyses showed no overall survival benefit but only a delay in myeloma progression. Initial treatment with two successive transplantation procedures has a negative risk benefit balance. (4) The optimal chemotherapy regimen prior to autologous stem cell transplantation is controversial. It is unclear which combination (vincristine + doxorubicin + dexamethasone, cyclophosphamide + dexamethasone, cyclophosphamide + dexamethasone, or bortezomib + dexamethasone, etc.) has a better risk-benefit balance in terms of survival and quality of life; (5) According to a meta-analysis of three clinical trials, thalidomide maintenance therapy appears to improve overall survival after Autologous stem cell transplantation; (6) Despite their inadequate evaluation, lenalidomide and pegylated liposomal doxorubicin are licensed for use in patients who relapse or who are refractory to initial treatment. In view of their major adverse effects, we consider that these drugs should only be used in clinical trials. PMID- 20025099 TI - Forensic science and human rights: the challenges. PMID- 20025100 TI - Complex regional pain syndrome: the role of the psychiatrist as an expert witness. AB - Complex regional pain syndrome (CRPS) is a regional pain syndrome of unclear pathophysiology, typically affecting the hand or foot. CRPS can be subject to personal injury litigation and may be the cause of substantial future loss of earnings claims. In clinical practice, a variety of medical experts can be involved in the diagnosis and management of CRPS and this can lead to confusion about the most appropriate expert for lawyers to instruct. This paper defines the role of psychiatrists as experts to the court in CRPS and reviews the associated psychiatric literature. Psychiatrists do not have expertise in the clinical diagnosis of CRPS, but they are able to assess the presence of co-morbid psychiatric conditions. They can assist the court as experts in the underlying neuro-physiological and psychological processes which may explain some of the important features of CRPS and an individual's psychological vulnerability to CRPS. Their expertise in psychological and psycho-pharmacological interventions should be considered for any multidisciplinary treatment programme for CRPS along with the likely contribution of these interventions to the overall prognosis. PMID- 20025101 TI - Outcome of high security patients admitted to a medium secure unit: the Edenfield Centre study. AB - This study examined the outcome of a cohort of 72 patients from a high secure psychiatric hospital (HSPH) who were admitted to and discharged from the Edenfield Centre over a 16-year period. The sample was predominately male and mentally ill and the majority was subject to restriction orders. Compared with two similar studies focusing specifically on the outcomes of HSPH patients in medium security, our sample had fewer women and the group had more extensive criminal and psychiatric histories. None of our cohort was discharged to the community via district services or transferred to other hospitals. We found that the Edenfield Centre had relatively higher rates of readmissions to the HSPH and a slightly higher reconviction rate than similar studies focusing solely on the outcomes of HSPH patients. In line with previous research, those detained under the legal category of psychopathic disorder were more likely to be reconvicted following discharge to the community than those detained under mental illness. Further large scale studies are needed to compare the outcomes of HSPH patients discharged through medium secure services that operate parallel or integrated aftercare service models. PMID- 20025102 TI - Concentration distributions of the drugs most frequently identified in post mortem femoral blood representing all causes of death. AB - Interpreting the concentrations of drugs determined in post-mortem blood is not an easy task owing to poly-drug use, adverse drug-drug interactions, as well as a host of pre-analytical factors and various artefacts in post-mortem toxicology. Highly sensitive and specific methods (GC-FID, GC-NPD. GC-MS and LC-MS) were used to determine the concentrations of drugs in femoral blood from 24,876 autopsies representing all causes of death. Ethanol topped the list of psychoactive substances (N=8108 or 33%) at mean, median and highest concentrations of 1.43 g/L, 1.20 g/L and 8.0 g/L, respectively. In second place was paracetamol (N=2741 or 11%). Amphetamine and cannabis were the major illicit drugs at 13th and 15th positions, respectively. Newer antidepressants, citalopram (no 3), sertraline (no 14), venlafaxine (no 16) were prominent as were sedative-hypnotics, such as diazepam (no 4), zopiclone (no 5) and zolpidem (no 18). This compilation of drugs and their concentration distributions will be useful to identify and flag for a likely overdose or drug-related poisoning death. The drug concentration together with the findings at autopsy and the police report can then be used to reach a conclusion about the cause and manner of death. PMID- 20025103 TI - Genetics and criminal behaviour: recent accomplishments. AB - The past two decades have seen an explosion in research in the fields of violence and behavioural genetics. Advances in human genetics have raised the possibility that genetic mechanisms can explain various aspects of human criminal and aggressive behaviour. However, this new knowledge can pose enormous challenges concerning the moral and legal conceptions of free will and responsibility. This paper reviews the main aspects of behavioural genetics, focusing on criminal and aggressive behaviour and describes the most important genes known to influence this behaviour. PMID- 20025104 TI - True and simulated homicidal hangings: a six-year retrospective study. AB - The manner of death in hangings is virtually always suicide. Uncommonly, accidental hangings do also occur. Homicidal hangings, however, are generally thought to be highly unusual. Despite some case reports of homicidal hangings, retrospective studies have demonstrated that homicidal hangings are virtually non existent. The present study was undertaken retrospectively to evaluate the incidence of true and simulated homicidal hangings in the forensic population of Quebec (Canada) during a six-year study period. In a total of 251 cases of hanging, suicide was the leading manner of death (239 cases); the remaining cases were accidents (eight cases) and homicides (four cases). This unusually high homicidal hanging rate in Quebec (1.6%) is hard to explain. It could be attributed to an intrinsic particularity of the population and culture. It could also be that our forensic team shares a strong interest in asphyxial deaths as a research topic and, therefore, is particularly alert in detecting such homicides. Nevertheless, this relatively higher homicidal rate may be a reminder that homicidal hangings do sometimes occur. This study emphasises the importance of not disregarding this manner of death in hangings. PMID- 20025105 TI - Service audit of a forensic rehabilitation ward. AB - An open forensic rehabilitation ward provides an important link bridging the gap between secure and community provisions. This paper provides an audit of such a service by examining the records of an open forensic rehabilitation ward over a five-year period from 1 June 2000 until 31 May 2005. During the audit period there were 51 admissions, involving 45 different patients, and 50 discharges. The majority of the patients came from secure unit facilities, acute psychiatric wards or home. Thirty-nine patients were discharged either into hostels (66%) or their home (12%). The majority of patients (80%) had on admission a primary diagnosis of either schizophrenia or schizoaffective disorder. Most had an extensive forensic history. The focus of their admission was to assess and treat their mental illness/disorder and offending behaviour and this was successful as the majority of patients were transferred to a community placement after a mean of 15 months. It is essential that there is a well-integrated care pathway for forensic patients, involving constructive liaison with generic services and a well-structured treatment programme which integrates the key principles of the 'recovery model' approach to care. PMID- 20025106 TI - An unrecognised case of withdrawal neuroleptic malignant syndrome: a case report. AB - Withdrawal neuroleptic malignant syndrome is a not widely recognised variant of the rare adverse consequence of neuroleptic medication, neuroleptic malignant syndrome (NMS). We present a case to bring the disorder to the attention of clinicians responsible for treating serious mental illness and medical and judicial officers who may be involved in investigating serious untoward incidents occurring within a mental health care setting. PMID- 20025107 TI - Suicide by burning barbecue charcoal: three case reports. AB - We report three cases of suicide in Scotland where barbecue charcoal was purposely burned in confined areas (an outbuilding, a car and a bedroom). External examination of the three cases revealed a distinctive 'cherry red' discolouration to the post-mortem lividity and blood and there were no marks or injuries to the bodies to give any cause for concern. Toxicological analysis of femoral blood samples revealed fatal levels of carbon monoxide (70%, 85% and 80% respectively). Considering the history, circumstances and external findings, a 'View and Grant' examination was conducted in all three cases and the cause of death was attributed to carbon monoxide poisoning due to inhalation of burning charcoal fumes. This particular method of suicide is not common in Europe and is more widely reported in the Far East. PMID- 20025108 TI - Fatal blunt injuries possibly resulting from sexual abuse of a calf: a case report. AB - We report a case of a calf-related fatality on a farm. The victim was a 60-year old man who was possibly attempting to sexually approach the animal from behind when he was struck by it, sustaining fatal blunt injuries, mostly to the thorax. This case is reported because of its rarity and because of the particular circumstances of death. Fatal trauma associated with bovines is rare on the whole, and when it does occur it is usually caused by kicking or pushing during regular agricultural activity. Although the cause of death in our case could be determined, the circumstances and the manner of death remain speculative to some degree. PMID- 20025109 TI - 2008 C. H. McCloy lecture. Social psychology and physical activity: back to the future. AB - In the early 1970s, both my academic career and the psychology subdiscipline within kinesiology began as "social psychology and physical activity. "Since then, sport and exercise psychology research has shifted away from the social to a narrower biopsycho-(no social) approach, and professional practice has focused on the elite rather than the larger public. Psychology can contribute to an integrative and relevant professional discipline by going back to the future as social psychology and physical activity and by incorporating three of C. H. McCloy's themes (a) evidence-based practice, (b) beyond dualisms, and (c) commitment to public service. Our scholarship must move beyond dualisms to recognize complexities and connections and be truly scholarship for practice. Social psychology and physical activity can serve the public by advocating for inclusive, empowering physical activity programs that promote health and well being for all. PMID- 20025110 TI - Research Quarterly for Exercise and Sport lecture. Statewide physical fitness testing: a big waist or a big waste? AB - Statewide physical fitness testing is gaining popularity in the United States because of increased childhood obesity levels, the relations between physical fitness and academic performance, and the hypothesized relations between adult characteristics and childhood physical activity, physical fitness, and health behaviors. Large-scale physical fitness testing can be fraught with problems unless properly planned and conducted. Legislators, administrators, teachers, and parents should consider the following 10 essential issues when conducting large scale physical fitness testing purpose of testing, proper planning, training, quality of the data, reporting support, costs, interpretation, programmatic matters, and policies and politics. PMID- 20025111 TI - One- or two-legged standing: what is the more suitable protocol to assess the postural effects of the rigid ankle orthosis? AB - To highlight the capacity of one- and two-legged standing protocols when assessing postural behavior induced by a rigid ankle orthosis, 14 healthy individuals stood upright barefoot and wore either an elastic stocking on the preferred leg or a rigid orthosis with or without additional taping in one- or two-legged (TL) conditions. Traditional center-of pressure (CP) measures were evaluated for the total two-feet resultant CP and under the feet (plantar CP). Focusing on the plantar CP displacements under the leg fitted with the various orthoses demonstrated particular postural behaviors for traditional parameters with main effects along the mediolateral axis. Only the TL protocol showed the limiting effects of the rigid shells on the inversion-eversion movements in healthy individuals. PMID- 20025112 TI - Focus groups to explore the perceptions of older adults on a pedometer-based intervention. AB - Focus group methodology was used to explore in depth the perceptions of older adults who had participated in a 12-week pedometer-based intervention. Nineteen women and 8 men, ages 55-86 years, volunteered to take part in the focus groups following participation in the intervention. Four focus groups of six to eight participants were scheduled at primary sites in the southern Maine area. Analysis of the data revealed four specific topical areas: (a) factors that led to increases in daily step count, (b) factors that hindered increases in daily step count, (c) benefits of pedometer intervention program, and (d) recommendations on how to improve older adults' physical activity. Overall, focus group participants found this intervention to be beneficial and recommended its expansion to other groups. PMID- 20025113 TI - Free-throw shooting during dual-task performance: implications for attentional demand and performance. AB - In this study, the dual-task paradigm was used to determine peak attentional demand during the free-throw process. Thirty participants completed 40 free-throw trials. The free throw was the primary task, but participants also verbally responded to a tone administered at one of four probe positions (PP). Repeated measures analysis of variance showed no significant difference in free-throw performance across PPs, indicating participants were able to keep the free throw as the primary task. Repeated measures analysis of response time (RT) showed significant differences, with RT at PP1 (preshot routine) and PP2 (first upward motion of the ball) significantly higher than baseline RT These results suggest that PP1 requires the greatest attentional demand, followed by PP2. PMID- 20025114 TI - Attentional mechanisms in sports via brain-electrical event-related potentials. AB - In this study, we examined attention processes in complex, sport-specific decision-making tasks without interdependencies from anticipation. Psychophysiological and performance data recorded from advanced and intermediate level basketball referees were compared. Event-related potentials obtained while judging game situations in foul recognition and a control task provided insight into focus of attention, selective attention, and processing strategy (top-down vs. bottom-up). Results showed task-specific effects for advanced referees in components influenced by attentional focus and selective attention. Experts also seemed to profit from superior top-down strategy and were able to evaluate the stimuli more rapidly. These findings are discussed in connection with current models in neurosciences and theories of referee research. PMID- 20025115 TI - Cortisol response to physical activity in African American toddlers attending full-time day care. AB - The purpose of this study was to examine African American toddlers' cortisol response to acute physical play activity within a full-time subsidized day care environment. Saliva samples were taken from participants (N = 22, ages 26-45.5 months) before and after physical play and control play conditions at the same time of day. Actiheart monitors were used to estimate the intensity of the play conditions. Although heart rate and other indicators were significantly higher during physical play, no change in cortisol levels was seen pre- to postphysical play. Further research is needed to better understand the cortisol response observed. PMID- 20025116 TI - Urban minority ninth-grade students' health-related fitness knowledge. AB - The purpose of the study was to examine Hispanic and other minority ninth-grade students' health-related fitness (HRF) knowledge, using FitSmart, a standardized test for high school students. The test consisted of 50 weighted multiple choice items, measuring six subcontent components. Means and standard deviations of the overall scores and the subcontent scores were computed. The findings suggested that students demonstrated mastery of one subcontent component but failed to meet passing values for all other subcontents and overall HRF knowledge. Female students outscored their male counterparts, but there was no difference for ethnicity. More efforts are needed to ensure that Hispanic and other minority high school students master fundamental HRF knowledge. PMID- 20025117 TI - Evaluation and opportunities in overtraining approaches. AB - Overtraining (OT) as a sports phenomenon can be caused by stressors on various levels (physical, emotional, psychological, and social) and evokes responses on these levels. This study evaluated research and new opportunities in the field of OT by introducing an integrated multidisciplinary approach, based on the single and multistressors approach. The single stressor approach focuses on the training load-recovery imbalance, which results in a stagnating performance, excluding the etiology by nonsport-related factors. The multistressors approach includes all factors as relevant in the etiology of a stagnating performance. In future studies on OT an integrative approach should not only highlight changes in training regimes and specific responses to training stressors but also focus on the role of training-related recovery, the impact of stressors, and personality factors influencing stress appraisal. This will provide a better insight into the etiology and consequences of OT necessary for prevention and treatment in sport practice, and enhance the focus on adequate recovery (good sleep, sufficient rest periods) and athletes' stress-related responses. PMID- 20025118 TI - Peers and teachers as sources of relatedness perceptions, motivation, and affective responses in physical education. AB - Research has demonstrated the importance of relatedness perceptions to self determined motivation in physical education. Therefore, studies have begun to examine the social factors contributing to feelings of relatedness. The purpose of this study was to examine teacher (perceived emotional support) and peer (acceptance, friendship quality) relationship variables to feelings of relatedness, motivation, and affective responses in junior high physical education students (N = 411). Results revealed that perceived relatedness mediated the relationship between variables and self-determined motivation and related directly to the amount of enjoyment and worry students experienced. These findings demonstrate that relationships with both teachers and peers are important for students' relatedness perceptions, motivation, enjoyment, and worry in physical education. PMID- 20025119 TI - Psychological need fulfillment among workers in an exercise intervention: a qualitative investigation. AB - The purpose of this study was to explore the perceived factors affecting workers'participation in an exercise intervention and interpret the findings within self-determination theory (Ryan & Deci, 2000a; 2007). Research examining the impact of psychological need satisfaction on exercise outcomes is not well established (McDonough & Crocke, 2007; Ryan & Deci, 2007). Even less is known about the processes through which workers negotiate a range of individual, group and context-based factors to fulfill these basic needs in exercise settings. To provide such insight, focus group interviews were conducted with 10 factory workers to discuss their involvement in a 7-week exercise intervention. Results indicated that a negotiation of complementary and competing factors (i.e., skill acquisition and regaining physical capabilities, a sense of camaraderie, characteristics of the trainer exercise context, a sense of obligation, scheduling) affected workers' need fulfillment, which ultimately affected their exercise adherence. The implications for health practitioners aiming to meet the psychological needs of workers involved in exercise interventions are discussed. PMID- 20025120 TI - The impact of an obstacle course sport education season on students' aerobic fitness levels. PMID- 20025121 TI - A survey of basic instructional program graduate teaching assistant development and support processes. PMID- 20025122 TI - Structural implications of the EL(KIQ)(L/C)LD(A/G)DD sequence in the C-terminal cytoplasmic tail for proper targeting of anion exchanger 1 to the plasma membrane. AB - While the C-terminal cytoplasmic tail of anion exchanger 1 (AE1, band 3) has been reported to possess important physiological roles, including one for proper membrane trafficking, its precise characteristics remain unclear. To clarify the overall structural consequences of the conserved sequence EL(K/Q)(L/C)LD(A/G)DD, containing the core binding sequence LDADD for carbonic anhydrase II, in the C terminal region, we analyzed the membrane expression and turnover of bovine AE1 with a series of truncation and substitution mutations in HEK293 cells. Immunofluorescence microscopy and cell-surface biotinylation demonstrated that truncation mutants missing 18 C-terminal residues targeted the plasma membrane, but the one lacking the conserved region, by truncation of 28 amino acid residues, was retained inside the cells. Substitutions of Ala for Glu901, Leu902, Leu905, and Asp906 in the sequence E901L(K/Q)(L/C)LDADD909 of bovine AE1 or those in the corresponding murine sequence also caused intracellular retention, though these mutants had half-lives comparable to that for wild-type AE1. These data demonstrate that the conserved amino acid residues Glu1, Leu2, Leu5, and Asp6 in the EL(K/Q)(L/C)LD(A/G)DD region have essential structural consequences in stable expression of AE1 at the plasma membrane regardless of the ability in binding to carbonic anhydrase II of this region. PMID- 20025123 TI - Growth variation in skull morphology of Kuril harbor seals (Phoca vitulina stejnegeri) and spotted seals (Phoca largha) in Hokkaido, Japan. AB - We examined morphological growth variations in skull features between the Kuril harbor seal and the spotted seal in Hokkaido, Japan. Skulls from 80 Kuril harbor seals and 41 spotted seals were collected, and we measured 29 metric and 6 non metric cranial characteristics. Three growth classes were defined according to the postnatal developmental stage: pups (0 year), subadults (1-4 years old) and adults (more than 5 years old). We detected sexual dimorphism in Kuril harbor seal pups, subadults, and adults. Although interspecies differences were detected in each growth class, Kuril harbor seals were larger and more massive than spotted seals; this feature was already detectable in pups. We did not detect certain cranial characteristics with which to identify the two species, but it was possible to identify any unknown specimens to their species, sex, and growth class using the cranial data generated in this study. Using 6 non-metric cranial characteristics, we identified significant interspecies differences with regard to the shape of the temporozygomatic suture and the extent of the nasal-incisive suture; the shape of the temporozygomatic suture and the shape of the nares were indicators of growth class in Kuril harbor seals. Although non-metric cranial characteristics have a lower discriminating power than metric characteristics, they are easy to use in the field even by inexperienced researchers. PMID- 20025124 TI - Proinsulin C-peptide induces c-Jun N-terminal kinase 1 expression in LEII mouse lung capillary endothelial cells. AB - To characterize the roles of C-peptide in vascular homeostatic processes, we examined the genes regulated by C-peptide in LEII mouse lung microvascular endothelial cells. Treatment of the cells with C-peptide increased the expression of c-Jun N-terminal kinase 1 (JNK1) mRNA dose-dependently, accompanied by an increase in JNK1 protein content. Prior treatment of the cells with PD98059, an ERK kinase inhibitor or SB203580, a p38MAPK inhibitor, abrogated the C-peptide elicited JNK1 mRNA expression. These results indicate that C-peptide increases JNK1 protein levels, possibly through ERK- and p38MAPK-dependent activation of JNK gene transcription. PMID- 20025125 TI - Radioecological investigation of food of animal origin in Belgrade environment. AB - The activity concentrations of 40K, 238U, 232Th and 137Cs were measured using gamma spectrometric method in different food chain samples from Serb Belgrade environment during the periods May-June 2007 and May-June 2008 year. Relatively high activities of 40K and 137Cs were detected in the soil. These results indicate that 137Cs is present in Belgrade environment even 20 years after nuclear accident in Chernobyl. However, in the samples of feedstuffs, animal products and bio indicators (meat of wild animals and fish), activity concentrations of primordial radionuclides and 137Cs were low and below the detection limits. Results of these trials have shown that investigated animal products from the natural environment around Belgrade, are radioactivity safe. PMID- 20025126 TI - [Dear Authors and Dear Readers!]. PMID- 20025127 TI - New vistas and views in the concept of generalized epilepsies. AB - The aim of this work is to show explicitly why the "idiopathic generalized epilepsy" concept becomes outfashioned and untenable. As the concept of "generalized epilepsies" is from long ago closely related to the thalamo-cortical system, we briefly summarize the functional anatomy, the double working mode of the thalamo-cortical system in different vigilance states and it's role in development of the spike-wave pattern. The next part shows weaknesses of this concept from the EEG, seizure semiology, and neuroimaging point of view. Further experimental and clinical arguments are accumulated from the reflex epileptic features in IGE, indicating local/regional cortical hyperexcitability. A separate part is devoted to genetic aspects of the question. Lastly implications to epilepsy classification are shown and an outlook toward a unified epilepsy concept is provided. The epileptic disorder of the thalamo-cortical system is responsible for the development of "generalized", synchronous spike-wave paroxysms as the common neurophysiological background in "primary" - idiopathic and in "secondary" generalized epilepsies. This disorder is specifically related to the burstfiring working mode of the thalamo-cortical system during NREM sleep (is an epileptic exageration of it). The "generalized" epilepsy category should be abandoned, being misleading. Epilepsies are proposed to be classified according to their network properties and relations to different physiological systems of the brain. The different phenotypes, named earlier idiopathic (primary) generalized, or symptomatic (secondary) generalized (with encephalopathic features), should be delineated depending on the following factors: 1. speed and extent of syncronization within the thalamo-cortical system, 2. the way how the thalamo-cortical system is involved, 3. which kind of cortical triggers play role, 4. the degree and level of the disorder (restricted to the molecular level or extended to the level of structural alterations - in the cortex or more diffusely, 5. genetic targets and features. PMID- 20025128 TI - [Role of zonisamid in treating epilepsy, Parkinson disorders and other neurological diseases]. AB - On the basis of six randomized controlled trials, zonisamide (ZNS) can be prescribed as add-on treatment in focal adulthood epilepsy in USA and Europe. In Japan, it can be prescribed as first-line monotherapy drug - independent of age. ZNS may also be effective in idiopathic generalized epilepsy and some difficult to-treat epilepsies including West, Lennox-Gastaut, or Dravet syndromes. The most frequent side effects of ZNS are related to central nervous system occurring in 19%. Kidney stones and oligohidrosis are ZNS-specific side effects. Loss of appetite and weight are usually "beneficial" effects. ZNS is not recommended in pregnancy ZNS can be taken once daily, which may be beneficial in non-compliance. The pathomechanism of ZNS is different from other antiepileptic drugs. ZNS has an effect on the voltage-gated Na+ - and T-type Ca2+ channels as well as on the dopaminerg, glutamaterg, cholinerg, and GABAerg systems. The multiple way of action may be the reason why ZNS seems to be a broad-spectrum drug and beneficial in various neurological disorders. ZNS reduces production of free radicals according to in vitro and in vivo studies. Animal experiments suggest that ZNS may be a neuroprotective agent. Based on an adequate randomized controlled trial, ZNS is effective in adjuctive treatment of Parkinson disorder. A peculiar benefit of the ZNS is that parallel to its positive effect on motor impairment it also reduces severity of dyskinesias. ZNS may be effective in bipolar disorder, obesity, eating disorders, and migraine prophylaxis. PMID- 20025129 TI - [Quantitative analysis of the genes determining spinal muscular atrophy]. AB - Spinal muscular atrophy (SMA) is one of the most common autosomal recessive diseases, affecting approximately one in 10,000 live births and with a carrier frequency of approximately one in 35. The disease is caused by a deficiency of the ubiquitous protein survival of motor neuron (SMN), which is encoded by the SMN1 and SMN2 genes. Due to a single nucleotide polymorphism in exon 7, SMN2 produces less full-length transcript than SMN1 and cannot prevent neuronal cell death at physiologic gene dosages. On the other hand, the copy number of SMN2 affects the amount of SMN protein produced and the severity of the SMA phenotype. SMN gene dosage analysis can determine the copy number of SMN1 to detect carriers and patients heterozygous for the absence of SMN1 exon 7. This study provides copy number estimation of SMN1 gene by real-time PCR technique in 56 SMA type I., II., III. patients, 159 parents and healthy relatives and in 152 undefined SMA patients. Among the family members, 91 carriers have been detected and in 56 patients homozygous deletion of SMN1 exon 7 has been confirmed. Moreover, in 12 patients compound heterozygosity of SMN1 exon 7 mutation has been detected, thus providing the possible diagnosis of SMA. In 94 patients, copy number of SMN2 has also been evaluated and a good correlation has been found with the phenotype of the disease. Due to the genetic complexity and the high carrier frequency, accurate risk assessment and genetic counselling are particularly important for the families. These new results provide improvement of the diagnostic service in SMA in Hungary with focus on proper genetic counselling and possible enrolment of the patients in future therapeutic interventions. PMID- 20025130 TI - [Guillain-Barre syndrome in childhood]. AB - BACKGROUND: Guillain-Barre syndrome (GBS) is clinically well known since 1916. It can occur at any age. Its main characteristic is acute rapidly ascending flaccid paresis. It is a neuro-immunologic disorder with heterogeneous background. In Hungary we could not find reports about big paediatric population with GBS. PATIENT AND METHOD: We analysed retrospectively the data of 38 children diagnosed and treated with GBS at the Neurological Department of Paul Heim Children's Hospital or at the Paediatric Department of St. Laszlo Hospital from January 2000 till April 2008. We analysed the clinical characteristics, seriousness of clinical signs, laboratory results, and electrophysiological features of them as well documented the preceding illness. We observed the effectiveness of our treatment; we measured the speed and time of the healing process and documented the residual clinical signs. RESULTS: 35 children could be classified as having acute inflammatory demyelinating polyneuropathy (AIDP), 2 as having acute motor axonal neuropathy (AMAN) and 1 as Miller-Fisher syndrome. By those patients who at the very beginning did not show the characteristic clinical signs, electrophysiology helped in establishing the diagnosis. By one child spinal MRI with gadolinium supported our diagnosis. Those children, who lost their ambulation, got immunotherapy: intravenous immunoglobulin (IVIG) or plasmapheresis (PEX). Both method seemed to be effective. None of our patients died. All were cured. By five patients residual clinical symptoms could be found. CONCLUSION: The disease process, the relative incidence of each subtype of GBS is nearly similar to that in Western Europe and North America according to the literature. By the currently used immune therapy most of the pediatric patients recover fully within a short time. PMID- 20025131 TI - [Acute transverse myelitis in childhood]. AB - We reviewed the medical history, clinical signs, imaging studies, laboratory data and treatment effectiveness of our 10 patients presented with acute idiopathic transverse myelitis. We used the criteria of the Transverse Myelitis Consortium Working Group (2002). So we excluded all those cases by whom the cause of the inflammation could be detected (e. g. direct viral inflammatory disease, systemic autoimmune disease). Age of the patients at disease onset ranged from 3 to 15 years. The first clinical signs were pain in different locations, and urinary retention. Paraparesis or plegia reached its maximum within five days. By all patients spinal MRI and lumbar puncture were performed at admission. These results were interpreted together with the clinical signs, and therapy was started immediately. We used methylprednisolon pulse therapy. Within 10-30 days the patients started to walk. We have followed the children for 1.5-13 years. Few residual clinical signs were observed: by one child left sided spastic monoparesis persisted, by the other right sided latent monoparesis was stated, and by one partial urinary incontinence persisted. By the control spinal MRI persisting signal changes or atrophy were detected just by those two children who had residual clinical signs. In the follow-up period no clinical relapse occured. Neither did the brain or spinal MRI show new lesions. The quick diagnosis and the immediately started therapy determine mostly the clinical outcome of these children. We hope that our long follow-up period can help in better understanding the disease even in adult patients. In the future we try to join multicenter clinical studies. PMID- 20025132 TI - Myelination disturbance in a patient with hyperuricemia and hyperserotoninemia combined with 18q deletion syndrome. AB - We previously reported a male patient with an 18q21.3 deletion, hyperuricemia and typical symptoms of the Lesch-Nyhan syndrome who lacked hypoxanthine-guanine phosphoribosyl-transferase (HGPRT) deficiency. The patient developed progressive peripheral neuropathy in additon to his profound mental retardation and self injurious behavior. At the age of 23 years MR imaging revealed globally delayed myelination with relative sparing of the corpus callosum and frontal lobes. They were focal hyperintensities suggestive of gliosis. Multimodality evoked potentials found evidence of impaired central and peripheral conduction. Single photon emission computed tomographic (SPECT) imaging demonstrated left frontal hyperperfusion and under it a temporoparietal hypoperfusion. PMID- 20025133 TI - Application of robots at patients with paresis of the upper limb as a consequence of central motor neuron lesion for supporting physiotherapy during rehabilitation. PMID- 20025134 TI - [Darwin anniversaries and neurology]. PMID- 20025135 TI - Cord blood adipocytokines beyond adiposity in neonates? PMID- 20025136 TI - The epithelial gatekeeper against food allergy. AB - The rapid rise of allergic disorders in developed countries has been attributed to the hygiene hypothesis, implicating that increased environmental sanitation in early childhood may be associated with higher incidence of hypersensitivity. Intestinal epithelial barriers play a crucial role in the maintenance of gut homeostasis by limiting penetration of luminal bacteria and dietary allergens, yet allowing antigen sampling via the follicle-associated epithelium for generation of tolerance. However, this intricate balance is upset in allergic intestines, whereby luminal proteins with antigenic properties gain access to the subepithelial compartment and stimulate mast cell degranulation. Recent studies demonstrated that food allergens were protected from lysosomal degradation, and were transported in large quantities across the epithelium by binding to cell surface IgE/CD23 (FcepsilonRII) that prevented the antigenic protein from lysosomal degradation in enterocytes. IL-4 (a Th2-type cytokine) not only increased production of IgE from B cells, but also upregulated the expression of CD23 on intestinal epithelial cells. Further studies indicated that CD23 was responsible for the bidirectional transport of IgE across epithelium. The presence of IgE/CD23 opens a gate for intact dietary allergens to transcytose across the epithelial cells, and thus foments the mast cell-dependent anaphylactic responses. The understanding of the molecular mechanism responsible for epithelial barrier defects may be helpful in designing novel therapies to treat food allergy and other allergic diseases. PMID- 20025137 TI - Association of cord plasma leptin with birth size in term newborns. AB - BACKGROUND: Leptin is secreted from adipose tissue and plays an important role in obesity. Recent studies have shown that the relationship between Leptin and body fat mass may have ethnic differences. The purpose of our study was to investigate the relationship between venous umbilical cord plasma Leptin and anthropometric markers in term healthy Taiwanese newborns. METHODS: Umbilical venous plasma samples were obtained from 98 term neonates (48 males and 50 females) and leptin Levels were analyzed by enzyme-linked immunosorbent assay. RESULTS: Umbilical cord plasma Levels of leptin were significantly higher in the female neonates than in males (p<0.001). The large-for-gestationaL age and appropriate-for gestational age newborns had significantly higher Leptin cord plasma levels than the small-for-gestational age newborns (p<0.01 and p<0.05, respectively). In both male and female neonates, umbilical Leptin Levels showed significant positive correlations with birth weight and birth Length. Multiple Linear regression analysis revealed that birth weight was the only significant predictor of umbilical cord plasma Leptin levels in both male and female neonates. However, the slopes of the regressions between Leptin and birth weight in male and female neonates were not different. CONCLUSION: In Taiwanese healthy term neonates, leptin umbilical cord plasma Levels are associated with sex and birth weight of the neonate. The relationship between Leptin and birth weight may differ among different ethnic groups. These findings imply that the relationship between leptin and body fat mass may develop early in life. PMID- 20025138 TI - Risk factor analysis for late-onset neonatal hyperbilirubinemia in Taiwanese infants. AB - BACKGROUND: Neonatal hyperbilirubinemia is a recognized health risk for newborns. It can cause kernicterus and hearing impairment. Certain groups of infants who do not have significant jaundice during the first few days of life develop hyperbilirubinemia later. Because early discharge is a worldwide trend, prompt identification of this group of infants is of paramount importance in preventing complications. METHODS: The data used were derived from a medical center. A total of 523 term or near-term infants were enrolled in this study. All infants were scheduled for follow-up visits. In this study, late-onset neonatal hyperbitirubinemia was defined as a total bilirubin level greater than 15 mg/dL, or receiving phototherapy at 5-7 days. Newborns that had clinically significant jaundice within 96 hours of life were excluded from analysis. Univariate and multivariate logistic regressions were applied for statistical analysis. RESULTS: One hundred and eighty infants were included for data analysis. Thirty-nine (21.7%) had late onset hyperbilirubinemia. Exclusive breast-feeding and less body weight loss during the 1st day of life were both significant risk factors for late onset hyperbilirubinemia. However, early discharge was not associated with late-onset hyperbilirubinemia. CONCLUSION: Thirty-nine (21.7%) infants develop late hyperbilirubinemia at the age of one week. Exclusive breast-feeding and less body weight loss were significant risk factors. Follow-up visits are recommended for all infants, especially for those who have these risk factors. PMID- 20025139 TI - Efficacy of double and triple phototherapy in term newborns with hyperbilirubinemia: the first clinical trial. AB - BACKGROUND: Icterus is the most common medical problem in neonates and phototherapy is the standard treatment for neonatal hyperbilirubinemia. This study aimed to compare the efficacy and length of hospital stay of double and triple phototherapy in newborns with indirect hyperbilirubinemia. METHODS: In a clinical trial, 40 healthy term newborns (gestational age >37 weeks) with bilirubin > or =12 and > or =15 mg/dL in the second and third day, respectively, were randomly assigned to triple (n=20) or double phototherapy (n=20) groups. Total and direct serum bilirubin were measured at admission, 8, 16, and 24 hours after beginning of phototherapy, and every 12 hours thereafter until discharge (when bilirubin levels reached < or =10 mg/dL). RESULTS: There were 21 male (52%) and 19 female (48%) term neonates, with significant difference between patients' age, weight, and age of icterus. There was no significant difference in the rate of bilirubin decline between the two groups within the first 8, 16 and 24 hours. There was no significant difference in mean length of hospital stay between the two groups. Analysis of complications showed no difference between triple and double groups. CONCLUSION: Triple phototherapy failed to reduce total serum bilirubin and shorten length of hospitalization more rapidly than double phototherapy. PMID- 20025140 TI - The influence of neonatal intensive care unit design on sound level. AB - BACKGROUND: Excessive noise in nurseries has been found to cause adverse effects in infants, especially preterm infants in neonatal intensive care units (NICUs). The NICU design may influence the background sound level. We compared the sound level in two differently designed spaces in one NICU. We hypothesized that the sound level in an enclosed space would be quieter than in an open space. METHODS: Sound levels were measured continuously 24 hours a day in two separate spaces at the same time, one enclosed and one open. Sound-level meters were placed near beds in each room. Sound levels were expressed as decibels, A-weighted (dBA) and presented as hourly L(eq), L(max), L(10), and L(90). RESULTS: The hourly L(eq) in the open space (50.8-57.2dB) was greater than that of the enclosed space (45.9 51.7dB), with a difference of 0.4-10.4dB, and a mean difference of 4.5dB (p<0.0001). The hourly L(10), L(90), and L(max) in the open space also exceeded that in the enclosed space (p<0.0001). CONCLUSION: The sound level measured in the enclosed space was quieter than in the open space. The design of bed space should be taken into consideration when building a new NICU. Besides the design of NICU architecture, continuous monitoring of sound level in the NICU is important to maintain a quiet environment. PMID- 20025141 TI - Peritoneal dialysis in infants and children after open heart surgery. AB - BACKGROUND: Infants and children who undergo surgical repair of complex congenital heart diseases are prone to developing renal dysfunction. The purpose of this study was to investigate the risk factors associated with prolonged peritoneal dialysis (PD) and the mortality of pediatric patients with acute renal failure (ARF) after open heart surgery. METHODS: From June 1999 to May 2007, a total of 542 children underwent open heart surgery for congenital heart disease. Fifteen (2.8%) experienced ARF and seven (1.3%) required PD. The clinical and laboratory variables were compared between the survivor and non-survivor groups of ARF patients that needed PD. RESULTS: The non-survivors (n=3, 43%) had a Longer cardiopulmonary bypass time (154+/-21 vs. 111+/-8 minutes, p=0.012) and longer aorta clamping time (92+/-40 vs. 66+/-15 minutes, p=0.010) than the survivors (n=4, 57%). Before the PD, the pH and base excess of the arterial blood gas analysis in the survivors was much higher than that non-survivors (7.30+/ 0.04 vs. 7.16+/-0.10, p=0.039; -5.15+/-3.13 vs. -12.07+/-2.9mmol/L, p=0.031). Furthermore, the survivors had a shorter interval between the onset of ARF and the day the PD was begun (1.2+/-0.4 vs. 4.3+/-1.2 days, p=0.001), and shorter duration of PD (6.6+/-2.7 vs. 13.0+/-3.5 days, p=0.036) than non-survivors. CONCLUSION: Early intervention with PD is a safe and effective method for managing patients with ARF after open heart surgery. The cardiopulmonary bypass and aortic clamping duration, time of initiating PD, duration of the PD, sepsis, and relative complications may predict the prognosis of these patients. PMID- 20025142 TI - Importance of prevention and early intervention of adverse events in pediatric cardiac catheterization: a review of three years of experience. AB - BACKGROUND: In spite of advances in many noninvasive instruments for studying cardiac anomalies in children, cardiac catheterization (CC) is still an essential method for the precise calculation of cardiovascular hemodynamic status and for performing therapeutic interventions. Accordingly, all adverse events (AE) related to CC are a major concern to pediatric cardiologists. PATIENTS AND METHODS: A total of 220 children with congenital heart disease (CHD) who received cardiac catheterization in our tertiary care hospital between the period of January 2000 and December 2002 were studied. One hundred and thirty-eight patients were non-cyanotic CHD, 71 were cyanotic CHD and 11 were complex CHD. Diagnostic CC was performed in 138 patients and therapeutic CC in 82 patients. All AEs that occurred during the CC procedures were identified, recorded and managed at the scene. The severities of AE were further classified into minor, obvious and severe. RESULTS: AEs were observed in 41 patients, including 22 (10%) minor, 16 (7.27%) obvious and 3 (1.36%) severe AEs. The three severe AEs were cardiac tamponade, severe ventricular tachycardia and marked hypoxia-and bradycardia. None of the patients died. Therapeutic CC did not present a higher incidence of AE occurrence than diagnostic CC. However, a young age (p<0.0001), low body weight (p<0.0001) and cyanotic or complex CHD (p=0.01) appeared to be risk factors for obvious and severe AE. CONCLUSION: Although the complication of severe AE during CC may not be totally preventable, it is important to be aware of every early sign of AE and to initiate an effective intervention by a well trained resuscitation team. PMID- 20025143 TI - Assessment of growth from foot length in Taiwanese neonates. AB - BACKGROUND: Previous studies have demonstrated a positive correlation between foot length (FL) and birth body weight (BBW), birth body length (BBL), and head circumference (HC). However, there is no data on birth FL in Taiwan. The aim of this study was to evaluate FL measurement in Taiwanese neonates as a method of estimating other anthropometric indices. METHODS: In this retrospective study, we enrolled 256 babies born at our hospital and Kaohsiung Veterans General Hospital from 2003-2005. Medical records were reviewed for sex, BBW, BBL, HC, gestational age, and birth FL. Ill newborns, small-for-gestational-age babies, or those with poor birth footprints were excluded. FL at birth was measured from the center of the back of the heel to the tip of the big toe. Linear regression analysis was used to investigate the relation of FL to BBW and BBL. The intraclass correlation coefficient was used to assess inter-rater reliability. RESULTS: A total of 256 babies were reviewed. There were 136 male and 120 female neonates. The gestational age was 38.5+/-1.3 (mean+/-standard deviation) weeks, ranging from 35 42 weeks. The BBW was 3137+/-396g. The BBL was 51.1+/-2.1 cm. The HC was 33.5+/ 1.7 cm. The FL was 7.4+/-0.46 cm. The regression equation for BBW (y) on FL (x) was as follows: y=486.2+360.4x (p<0.001, r=0.421). The regression equation for BBL (y) on FL (x) was as follows: y=40.1+1.45x (p<0.001, r= 0.305). The regression equation for HC (y) on FL (x) was as follows: y=14.8+2.53x (p<0.001, r=0.423). FL showed excellent reliability, with an intraclass correlation coefficient of 0.965 (p<0.001). CONCLUSION: Our study demonstrated a significant degree of correlation between FL and BBW, BBL and HC. However, it did not reliably estimate BBW, BBL, or HC-the three anthropometric indices were weakly correlated (r<0.5) with FL. PMID- 20025144 TI - Two different tests for total bilirubin in neonates and infants. AB - BACKGROUND: The most readily available methods for testing serum total bilirubin in neonates are the capillary tube-directed optics color method and serum biochemistry. Because inconsistency between these two methods may cause confusion in clinical practice, this study was designed to quantify their differences. METHODS: In 46 neonates with clinical jaundice, total bilirubin was measured by two different methods, using a nonchemical photometric device and a laboratory analyzer. RESULTS: Differences in results between these two methods were statistically significant, especially when total bilirubin level exceeded 15 mg/dL. CONCLUSION: Clinicians should be aware of the differences between the two methods when making decisions in patient care. PMID- 20025145 TI - Linear scleroderma "en coup de sabre": initial presentation as intractable partial seizures in a child. AB - Linear scleroderma is a form of localized scleroderma characterized by sclerotic lesions distributed in a linear, band-like pattern. The "en coup de sabre" subtype of linear scleroderma is more often associated with systemic morbidity, including ocular, oral, and neurological abnormalities. Here, we report one patient with typical linear scleroderma "en coup de sabre" (LSCS). Initially, he presented with refractory partial seizures before the characteristic skins lesion on his head developed. This was a rare case with obvious brain parenchyma involvement. We did not prescribe medication but performed serial brain magnetic resonance imaging follow-up for the intraparenchymal lesion. The atrophic changes of the skin, face and brain remained the same, and his seizures had not worsened at the most recent follow-up. Parry-Romberg syndrome, a very similar condition, should be differentiated from LSCS. PMID- 20025146 TI - Pioneer in advocacy: the legacy of James J. Peters. PMID- 20025148 TI - Computerized classification of neurologic injury based on the international standards for classification of spinal cord injury. AB - BACKGROUND/OBJECTIVE: To develop and test a computer program to accurately score International Standards for Classification of Spinal Cord Injury (ISCSCI) examinations. METHODS: A computer program was written following the algorithm published in the ISCSCI 2003 manual. Two methods were used to test the computer program's accuracy. First, 94 hand-scored examinations representing 25 different patients were entered into the program. Differences between hand-scored examinations and computer-scored examinations were evaluated for accuracy. Second, all case examples from the ISCSCI manual were entered into the computer program to verify its accuracy. RESULTS: Of the 94 hand-scored examinations, the computer detected all incorrectly scored examinations. Furthermore, the computer scoring agreed with every examination's scores that were correctly calculated by hand. Of the hand-scored examinations, 10% of sensory total scores and 4% of motor scores were incorrect. For sensory level and motor level, there were errors in 9% and 26% of hand-scored examinations, respectively; 13% of hand-scored examinations had incorrectly assigned ASIA Impairment Scale classification. CONCLUSION: This study showed that the computer program we developed was effective in correctly scoring ISCSCI examinations and was able to detect errors in hand-scored examinations. PMID- 20025147 TI - Review of critical factors related to employment after spinal cord injury: implications for research and vocational services. AB - BACKGROUND/OBJECTIVE: Employment rates after spinal cord injury (SCI) vary widely because of discrepancies in studies' definition of employment and time of measurement. The objective of this study was to provide a comprehensive summary of the literature on employment rates, predictors of employment, and the benefits and barriers involved. METHODS: A search using the terms spinal cord injury and employment in the databases PubMed, PsycINFO, and MEDLINE. The search included a review of published manuscripts from 1978 through 2008. RESULTS: A total of 579 articles were found and reviewed to determine the presence of reported employment rates. Of these, 60 articles were found to include a report of employment rates for individuals with SCI. Results indicated that, in studies that examined paid employment, the average rate of any employment after SCI was approximately 35%. CONCLUSIONS: Characteristics associated with employment after SCI include demographic variables, injury-related factors, employment history, psychosocial issues, and disability benefit status. It is recommended that researchers studying employment after SCI use common outcome measures such as competitive employment rates, duration of employment, and job tenure. Empirical evidence is lacking in regard to the most effective methods of vocational rehabilitation among this population. Evidence-based supported employment practices seem to be the most applicable model for assisting persons with SCI in restoring meaningful employment. Controlled studies are needed to test this assumption. PMID- 20025149 TI - Effects of acute nitric oxide synthase inhibition on lower leg vascular function in chronic tetraplegia. AB - BACKGROUND/OBJECTIVE: To improve our understanding of the lower-leg vascular responses of nitric oxide synthase inhibition in persons with tetraplegia. PARTICIPANTS: Six people with chronic tetraplegia and 6 age-matched controls. METHODS: Lower-leg relative vascular resistance and venous volume variation were obtained by venous occlusion plethysmography and blood pressure by auscultation at baseline. Postintravenous infusion of the nitric oxide synthase inhibitor NG nitro-L-arginine-methyl-ester (1 mg x kg(-1) or placebo on separate days. RESULTS: At baseline in the group with tetraplegia compared with controls, mean arterial pressure and relative vascular resistance of the leg were significantly lower. After nitric oxide synthase inhibition, mean arterial pressure and lower leg vascular resistance were significantly elevated in both groups. There were no group or intervention differences in venous volume variation. CONCLUSION: These preliminary results suggest that nitric oxide synthase inhibition with 1 mg x kg( 1) N(G)-nitro-L-arginine-methyl-ester normalizes seated blood pressure and lower leg vascular resistance to control group baseline levels. PMID- 20025150 TI - Information needs of people with spinal cord injuries. AB - BACKGROUND/OBJECTIVE: To determine the information needs, level of Internet access, and current and preferred formats and sources of information of adults with spinal cord injuries (SCIs). Individuals with SCIs have a high lifetime risk for medical complications and other health conditions secondary to their injury. Many secondary conditions can be prevented or mitigated through appropriate self care and/or self-management. People with SCIs need timely, high-quality information about health and medical issues after discharge and throughout their lifetime to improve self-care and maximize quality of life. METHODS: A survey was administered as part of the third time point of a longitudinal research study on individuals with SCI. RESULTS: A total of 80.2% of the 277 respondents reported having Internet access. The most frequently selected format used currently and preferred by respondents for receiving SCI information was "Web pages/Internet". The top-ranked current and preferred source of SCI information was from a "Physician: SCI Expert/Rehabilitation Specialist". Respondents reported needing information on medical issues the most. Significantly higher percentages of individuals identified as "white" and with higher education levels had access to the Internet and ranked the selection of Web pages/Internet as their top choice. CONCLUSIONS: Results confirm that, although people with SCI prefer to receive SCI information from SCI experts, the Internet is a more accessible and more currently used source. Educational level and race predicted current and preferred use of the Internet for obtaining SCI information, suggesting that Internet distribution of SCI information will exclude subgroups. PMID- 20025151 TI - Flexion model simulating spinal cord injury without radiographic abnormality in patients with ossification of the longitudinal ligament: the influence of flexion speed on the cervical spine. AB - BACKGROUND/OBJECTIVE: It is suspected that the speed of the motion of the spinal cord under static compression may be the cause of spinal cord injury (SCI). However, little is known about the relationship between the speed of the motion of the spinal cord and its stress distributions. The objective was to carry out a biomechanical study of SCI in patients with ossification of the longitudinal ligament without radiologic evidence of injury. METHODS: A 3-dimensional finite element spinal cord model was established. After the application of static compression, the model underwent anterior flexion to simulate SCI in ossification of the longitudinal ligament patients without radiologic abnormality. Flexion of the spine was assumed to occur at 1 motor segment. Flexion angle was 5 degrees, and flexion speeds were 0.5 degrees/s, 5 degrees/s, and 50 degrees/s. Stress distributions inside of the spinal cord were evaluated. RESULTS: Stresses on the spinal cord increased slightly after the application of 5 degrees of flexion at a speed of 0.5 degrees/s. Stresses became much higher at a speed of 5 degrees/s and increased further at 50 degrees s. CONCLUSIONS: The stress distribution of the spinal cord under static compression increased with faster flexion speed of the spinal cord. High-speed motion of the spinal cord under static compression may be one of the causes of SCI in the absence of radiologic abnormality. PMID- 20025152 TI - Therapists' roles in pressure ulcer management in persons with spinal cord injury. AB - BACKGROUND: Among veterans with spinal cord injury (SCI), severe pressure ulcers (PrU) are treated by interdisciplinary rehabilitation teams in SCI units. METHOD: Cross-sectional survey administered to therapists attending a conference of the Therapy Leadership Council in SCI. PARTICIPANTS: Respondents included physical therapists (PTs; n=24) and occupational therapists (OTs; n=15). MAIN OUTCOME MEASUREMENTS: Wound care practices as indicated by 75% or more of participants as "usual practice". RESULTS: In general, therapist involvement with wound care was initiated by physician order (e.g., electrical stimulation) or postsurgery protocols. "Usual practice" after tissue healing included progressive range of motion; initial remobilization (first sitting after wound healing); progression of sitting time including assessment of skin tolerance; instruction in pressure relief maneuvers/techniques; and instruction in safe transfers. Practices in prevention of a new ulcer included education and evaluation of seating posture/positioning. CONCLUSIONS: Results indicate that centers may delegate responsibilities for management of ulcers differentially by discipline. A limitation was that we were unable to determine whether these centers were the same or different for OT and PT respondents. Although sample size was small and some sites had multiple respondents, the survey showed a growing role for OTs and PTs in PrU treatment. Because 75% of each discipline reported that there were usual practices, including patient education and remobilization protocols, this area requires further study to determine the clinical outcomes in terms of preventing PrUs and recurrence. PMID- 20025153 TI - Validity, accuracy, and predictive value of urinary tract infection signs and symptoms in individuals with spinal cord injury on intermittent catheterization. AB - BACKGROUND/OBJECTIVE: To determine the validity, accuracy, and predictive value of the signs and symptoms of urinary tract infection (UTI) for individuals with spinal cord injury (SCI) using intermittent catheterization (IC) and the accuracy of individuals with SCI on IC at predicting their own UTI. DESIGN: Prospective cohort based on data from the first 3 months of a 1-year randomized controlled trial to evaluate UTI prevention effectiveness of hydrophilic and standard catheters. PARTICIPANTS: Fifty-six community-based individuals on IC. MAIN OUTCOME MEASURES: Presence of UTI as defined as bacteriuria with a colony count of at least 10(5) colony-forming units/mL and at least 1 sign or symptom of UTI. METHODS: Analysis of monthly urine culture and urinalysis data combined with analysis of monthly data collected using a questionnaire that asked subjects to self-report on UTI signs and symptoms and whether or not they felt they had a UTI. RESULTS: Overall, "cloudy urine" had the highest accuracy (83.1%), and "leukocytes in the urine" had the highest sensitivity (82.8%). The highest specificity was for "fever" (99.0%); however, it had a very low sensitivity (6.9%). Subjects were able to predict their own UTI with an accuracy of 66.2%, and the negative predictive value (82.8%) was substantially higher than the positive predictive value (32.6%). CONCLUSIONS: The UTI signs and symptoms can predict a UTI more accurately than individual subjects can by using subjective impressions of their own signs and symptoms. Subjects were better at predicting when they did not have a UTI than when they did have a UTI. PMID- 20025154 TI - Role of conventional and diffusion-weighted magnetic resonance imaging of spinal treatment protocol for hydatid disease. AB - OBJECTIVE: To demonstrate the role of magnetic resonance imaging (MRI) in determining the treatment protocol for hydatid disease of the spine. DESIGN: Case report; literature review. FINDINGS: Diffusion-weighted MRI can help differentiate complicated infected hydatidosis from abscesses, epidermoid cysts from arachnoid cysts, and benign from malignant vertebral compression fractures. It is also helpful in differentiating between abscesses and necrotic tumors. CONCLUSION: Diffusion-weighted MRI can help differentiate between infections requiring immediate surgery and those that can be treated medically with antihelmintic treatment. PMID- 20025155 TI - Balloon-tipped catheter for measuring urethral pressures. AB - BACKGROUND: Better methods are needed for recording urethral function for complex urologic problems involving the bladder, urethra, and pelvic floor. OBJECTIVE: To evaluate a balloon catheter for recording urethral pressure and function using bench-top testing and evaluation in an animal model. METHODS: Balloon pressure recording methods included slightly inflating the balloon with water and placing the pressure transducer on the distal end of the catheter. For bench-top testing, manual procedures and a silastic tube with a restriction were used. In 3 anesthetized dogs, pressure recorded from the skeletal urethral sphincter was induced with electrical stimulation of the sphincter. Anal sphincter pressure was also recorded. RESULTS: Bench-top testing showed good pressure recordings, including a confined peak at the tube restriction. Animal tests showed urethral pressure records with rapid responses when electrical stimulation was applied. Peak pressure at the urethral skeletal sphincter was 55.7 +/- 15 cmH2O, which was significantly higher than the peak pressure recorded 2 cm distally in the proximal urethra (3.3 +/- 2.3 cmH2O). Peak anal pressures were smaller and unchanged for the 2 stimulations. CONCLUSIONS: Balloon-pressure recordings showed rapid responses that were adequate for the tests conducted. In the animal model, high-pressure contractions specific to the skeletal urethral sphincter were shown. Balloon-tipped catheters warrant further investigation and may have applications for the evaluation of detrusor-sphincter dyssynergia after spinal cord injury or for stress urinary incontinence. PMID- 20025156 TI - Spinal glioblastoma multiforme: unusual cause of post-traumatic tetraparesis. AB - BACKGROUND/OBJECTIVE: Glioblastoma multiforme (GBM) is the most common glial cell tumor of the adult brain. However, primary GBM of the spinal cord is a rare condition. METHODS: Case report. RESULTS: A young man presented with acute onset quadriparesis after a whiplash injury. A magnetic resonance scan showed the typical appearance of a high-grade intramedullary tumor with fusiform expansion of the entire cervical cord. Subtotal decompression and biopsy was done by posterior laminectomy, followed by external beam radiotherapy. Signs and symptoms improved after the completion of radiotherapy but did not resolve completely. Death caused by respiratory failure occurred 3 months later. CONCLUSIONS: This presentation of GBM of the cervical cord is rare; an intramedullary tumor should be considered when minor cervical trauma results in disproportionate neurologic deficit. To the best of our knowledge, this is the first reported case of spinal GBM with extensive pan-cervical involvement. PMID- 20025157 TI - A vascular cause of painful lumbar transverse syndrome. AB - BACKGROUND: Despite typical signs of Leriche syndrome, this patient was misdiagnosed with disk prolapse. Vascular insufficiency, such as aortic occlusion, may underlie a motor-sensory deficit in the lower extremities. METHOD: Case report. FINDINGS: A 67-year-old woman was admitted for cramping pain in the lumbar spine, hips, and legs; diffuse weakness and sensory disturbances of both legs for weeks; and a cold feeling and livid colorization of the legs. Her complaints were attributed to a disk prolapse L5/S1, for which she underwent immediate laminectomy. Postoperatively, symptoms and signs were aggravated, abdominal pain developed, and C-reactive protein and creatinine were elevated. On the fourth postoperative day, femoral pulses were no longer palpable, anuria developed, and computed tomographry angiography showed occlusion of the abdominal aorta distal to the superior mesenteric artery and absent kidney perfusion. The patient died one day later. Postmortem review showed a visible intra-aortic mass on the previous lumbar magnetic resonance image. CONCLUSIONS: The differential diagnosis of painful lumbar transverse syndrome should include vascular disease. Misinterpretation of a Leriche syndrome as a disk prolapse has not been reported. PMID- 20025158 TI - Vision loss after spinal fusion for scoliosis in a child with spinal cord injury. AB - BACKGROUND: Vision loss after spinal surgery is a rare and devastating complication. Risk factors include patient age, operative time, estimated blood loss, and intraoperative fluid management. Children with spinal cord injury often develop scoliosis that requires surgical correction. STUDY DESIGN: Case report. METHODS: Clinical and radiographic review was conducted of a 15-year-old boy who developed severe scoliosis after sustaining a C5 level injury at age 4 years from a motor vehicle crash. FINDINGS: The patient underwent a posterior spinal fusion from T2 to the pelvis, and good correction of the spinal deformity was attained. During the 8-hour procedure, blood loss was 4000 mL (approximately 1.2 blood volumes) and 17,000 mL of fluids were administered. On postoperative day 5, it was determined that the patient had complete visual loss. Neuro-ophthalmology consultation confirmed the diagnosis of posterior ischemic optic neuropathy. CONCLUSIONS: A significant number of children with spinal cord injury develop scoliosis requiring surgical correction. These procedures are often lengthy, with the potential for extensive blood loss and fluid shifts, factors that may increase the likelihood of postoperative vision loss. Patients should be counseled about this complication, and the surgical and anesthesiology teams should take all measures to minimize its occurrence. PMID- 20025159 TI - Thoracic myelopathy secondary to intradural extramedullary bronchogenic cyst. AB - BACKGROUND/OBJECTIVE: To report a case of thoracic myelopathy secondary to intradural extramedullary bronchogenic cyst. STUDY DESIGN: Case report. METHODS/FINDINGS: A 20-year-old man presented to the emergency department with increasing back pain and lower-extremity weakness. Magnetic resonance imaging demonstrated a cystic lesion at the T4 level with mass effect on the spinal cord. RESULTS: The lesion was resected, and histopathologic evaluation showed a cyst lined by respiratory-type epithelium consistent with a bronchogenic cyst. CONCLUSIONS: Intradural extramedullary bronchogenic cysts of the thoracic spine have been reported previously but are extremely rare. The treatment of choice is surgical resection. PMID- 20025160 TI - Are you on board with the Joint Commission's FPPE/OPPE requirements? PMID- 20025161 TI - A new look at the hospital patient grievance process. PMID- 20025163 TI - Patient satisfaction is a best practice. PMID- 20025162 TI - Patient satisfaction planner. Patient-centric care decreases ED visits. PMID- 20025164 TI - TJC's take on patient complaints. PMID- 20025165 TI - Preparing for your 2010 survey? Here's some tips. PMID- 20025166 TI - [Infection with mycobacterium ulcerans (Buruli ulcer): still a neglected disease in 2009?]. PMID- 20025167 TI - [Alveolar echinococcosis in China: efficacy of long-term albendazole]. PMID- 20025168 TI - [Negative pressure therapy in precarious situations. Part 1: abdomen and perineum]. AB - Negative pressure therapy (NPT) is a proven safe and effective technique to promote healing of complex adominoperineal wounds using either an open or closed approach. Specifically designed devices are available in industrialized countries but high cost is a limiting factor for their use in developing countries. The purpose of this report is to describe a simple, easy-to-use technique that is suitable for austere environments. In addition to low-cost, the main advantage of this technique is that it can be performed by anyone using resources readily available in any operating room. PMID- 20025169 TI - [Melioidosis: an emerging tropical disease]. AB - Melioidosis is an infection affecting both human and animal health. The causative agent is Burkholderia pseudomallei, a Gram-negative soil bacterium. Melioidosis is endemic in tropical areas of Southeast Asia and Northern Australia, and sporadic in many other countries. Clinical presentation is variable ranging from acute septicemia, isolated pulmonary infection, or chronic granulomatous lesions to asymptomatic forms with positive serology. There is no vaccine and treatment is difficult because B. pseudomallei is resistant to a wide range of antibiotics. Relapses are common. B. pseudomallei is listed as a biological risk class 3 and considered as a potential bioterrorism agent due to its high virulence by inhalation, to the difficulty of treatment, and to the lack of vaccine. PMID- 20025170 TI - [Call for the revival of community medicine in Africa]. AB - In Africa persons seeking medical advice rarely see a physician first. The goal of initial contact is to ensure seamless interfacing with the other services of the healthcare system. The community physician in Africa is either a physician paid by contributions from people living in the region or, in most cases, a civil servant practicing at an outlying health center. Because community physicians often work in isolated locations, their colleagues at major hospital centers and in the academic world are rarely aware of their activity. As a result community physicians in Africa have little impact on the healthcare system. In fact their role should be to relieve overloading of hospitals that have become large dispensaries crowded with patients that have not received proper care in outlying facilities. In this article the authors call for community physicians in Africa to be brought out of isolation and given their true function as "clinicians with aptitudes in public health capable of assisting actions to restore and promote health and to prevent disease". PMID- 20025171 TI - [Sociocultural aspects of albinism in Sub-Saharan Africa: mutilations and ritual murders committed in east Africa (Burundi and Tanzania)]. AB - This report presents an overview of the many sociocultural prejudices confronting albinos and their parents in Sub-Saharan Africa at each stage of life (infancy, adolescence, and adulthood). The birth of an albino child to two black parents has always been an enigma for African peoples. French-speaking and English speaking populations in Central, East and South African countries have invented numerous myths to account for this event. Albinos are believed to possess good and evil magical powers. On the white magic side, some organs are believed to confer luck, health, and prosperity. For this reason albinos in Tanzania and Burundi are still in 2008 prey to ritual murders and mutilations to obtain various body parts such as arms, legs, and genitals for preparation of amulets. This barbaric and iniquitous practice has been severely condemned by authorities in both countries as well as by the European Parliament. To end these atrocities disseminating accurate medical information explaining the genetic basis of albinism will be necessary to eliminate ignorance and superstition. PMID- 20025172 TI - [Update on primaquine]. PMID- 20025173 TI - [HIV and sexually transmitted disease among female sex workers in Benin]. AB - BACKGROUND: This report describes a follow-up study conducted in 2005 to evaluate programs intended to reduce HIV/AIDS among registered and unregistered female sex workers (FSW) and their male clients in six major urban areas in Benin (Cotonou, Abomey-Bohicon, Parakou, Porto-Novo, Kandi, Malanville). The objectives of this second study combining laboratory testing and behavioral questionnaires were to estimate the prevalence of HIV and sexually transmitted diseases (STI), to determine the sociodemographic and behavioral characteristics of FSW, and to assess changes in the behavior of registered FSW in the first four cites (Cotonou, Abomey-Bohicon, Parakou, and Porto-Novo) included in the initial study in 2002. DESIGN AND METHODS: A cross-sectional study was conducted in a cohort of 930 self-identified FSW. The chi-square or Fisher's exact test were used to test correlation between HIV and social, demographic and behavioral factors and the Wilcoxon test to compare the distribution of continuous variables. Correlation was measured based on prevalence odd ratios (POR) obtained by multivariate logistic regression. RESULTS: The overall prevalence of HIV, Neisseria gonorrhoeae (NG) and Chlamydia trachomatis (CT) were 30.5%, 3.6% and 5.7% respectively. The prevalence of HIV was highest in Parakou (48.2%) and lowest in Abomey/Bohicon (16.4%). The prevalence of NG and CT were highest in Abomey/Bohicon and Porto-Nova respectively. Multivariate logistic regression analysis showed that HIV prevalence increased significantly for FSW who had had more than 10 clients in the previous week (POR = 1.40, 95% CI: 1.02-1.94), who had no other source of income (POR = 1.47, 95% CI: 1.08-2.00), who were currently unmarried or separated (POR = 2.63, 95% CI: 1.73-4.02), and who had never been married (POR = 2.07, 95% CI: 1.43-3.00). Older age and having had no non-paying partner in the last seven days were positively correlated with HIV infection. Between 2002 and 2005 there was a significant increase in the number of FSW who reported condom use with all clients (79% vs. 90%, p < 0.0001) and with all sexual partners (78% vs. 87%, p = 0.0007) during the last 7 days as well as in the median number of visits to a health care facility in the last 12 days (3 vs. 6, p < 0.0001). Overall these follow-up data showed a large decrease in the prevalence of HIV/STI. This finding supports continuation of programs aimed at FSW especially with regard to risk awareness, condom use, and STI screening and treatment. PMID- 20025174 TI - [Value of hand disinfection by rubbing with alcohol prior to surgery in a tropical setting]. AB - OBJECTIVE: The purpose of this study was to assess the value of hand disinfection by rubbing with alcohol in terms of prevention of surgical site infection (SSI) and cost in a tropical setting. MATERIAL AND METHOD: This prospective cohort study carried out in the Gynecology Obstetrics Department of the Yopougon University Teaching Hospital from May to September 2005 was designed to compare two different methods of hand disinfection, i.e., traditional 3-step hand scrubbing using an antiseptic preparation versus handwashing without an antiseptic preparation followed by two applications of an aqueous alcohol solution. RESULTS: The study population included 318 patients who underwent surgery during the study period. The SSI rate was 13.2% in patients operated on after traditional hand scrubbing and 11.5% after handwashing followed by rubbing with alcohol (not significantly different). Hand disinfection by rubbing with alcohol did not increase the risk of SSI and was considered as easier than traditional hand scrubbing by 90% of users. Skin tolerance was deemed good by 52% of users. Most users (69%) wished both hand disinfection methods to be available. Hand disinfection by rubbing with alcohol was much more cost-effective than traditional hand scrubbing both with regard to initial investment and to consumable costs (50% lower). CONCLUSION: This study shows that hand disinfection by rubbing with an alcoholic solution is not only as effective as traditional hand scrubbing for prevention of SSI but also more cost-effective. These findings indicate that rubbing with alcohol is a suitable alternative to traditional scrubbing for hand disinfection prior to surgery in our tropical setting. PMID- 20025175 TI - [Exophthalmia in children at the African Institute of Tropical Ophthalmology (AITO)]. AB - INTRODUCTION: Exophthalmia is a rare entity. The purpose of this study was to ascertain epidemiological features, clinical aspects, and potential treatment modalities for exophthalmia in children at the University Teaching Hospital- African Institute of Tropical Ophthalmology (UTH-AITO). PATIENTS AND METHOD: This prospective and descriptive study was carried out from July 2006 to June 2007 at the IOTA-UTH in collaboration with the pediatric hematology and oncology unit of the Gabriel Toure UTH. All patients between the ages of 0 to 15 years who presented unilateral or bilateral exophthalmia whether measurable or not were included. In all cases thorough clinical workup was performed and any appropriate additional examinations were ordered. All data were consigned to a questionnaire. RESULTS: A total of 46 eyes in 38 children were included, i.e., unilateral right in 15 cases, unilateral left in 15 and bilateral in 8. Mean age was 9.6 years (range, 2 days to 15 years). The male-to-female sex ratio was 1.9:1. The incidence of exophthalmia was 0.3%. Exophthalmia was the reason for seeking medical advice in 68.4% of cases (26 children). Classification according to underlying pathology divided patients into four groups, i.e., inflammatory and/or infectious pathologies in 36.9% (14 cases), tumor pathologies in 31.6% (12 cases), trauma in 13.1% (5 cases) and undetermined in 18.4% (7 cases). A Hertel millimetric regression of exophthalmia was observed in 23.9% of cases (11 eyes) and a gain in visual lines of acuity was observed in 8.7% (4 eyes). DISCUSSION: Given the normally low frequency of exophthalmia, observation of a 0.3% incidence at a third level reference center warrants alerting of health authorities. Etiological classification was comparable to Crawford's as described by Desjardins. Cellulites (50%) and retinoblastoma (33.3%) were the main causes of exophthalmia. CONCLUSION: Since exophthalmia can cause loss of vision and even be life-threatening, early diagnosis and treatment is important particularly in children. PMID- 20025176 TI - [Relapse after surgical treatment of mycobacterium ulcerans infection (buruli ulcer): study of risk factors in 84 patients in the Democratic Republic of the Congo]. AB - OBJECTIVE: To identify risk factors for relapse after exclusively surgical treatment of Mycobacterium ulcerans infection (Buruli ulcer). METHODS: Study was carried out in 102 patients treated exclusively by surgery for Buruli ulcer at various care facilities in the Congo from January 1, 2000 to January 1, 2005. RESULTS: Outcomes included relapse in 22 patients (21.5%), cure in 62 (60.7%), and unknown in 18 (17.6%). Statistical analysis identified the following variables as independent risk factors for relapse after exclusively surgical treatment: incomplete surgical excision (OR = 91.83; P = 0.0000; IC to 95%), age under 16 years (OR = 14.80; P = 0.0000; IC to 95%) and pre-ulcerative Buruli lesions (edema and plaque) (OR = 3.18; P = 0.0215; IC to 95%). CONCLUSION: Quality of excision, patient age, and clinical form of lesion are the main predictors of relapse after isolated surgical treatment of Buruli ulcer. PMID- 20025177 TI - [Drainage of liver abscess by "mini-hepatotomy"]. AB - We have developed a a "mini-hepatotomy" technique to prevent rupture of large superficial liver abscesses. AIM: The purpose of this report is to describe our results using this technique. PATIENTS AND METHOD: Data was extracted from the files of 40 patients hospitalized in our department for liver abscess from January 2000 to June 2005. Antimicrobrial therapy including metronidazole was administered to all patients and was the sole treatment in two. Additional treatment included laparotomy for ruptured abscess in 4 cases, pleural drainage for rupture in the right pleural cavity in one, and "mini hepatotomy" in 33. This study includes the 33 patients who underwent mini-laparotomy. There were 30 men and 3 women with a mean age of 38 years. The mean delay for consultation was 51 days. RESULTS: Liver abscess was classified as amoebic abscess in 28 cases and pyogenic abscess in 5. The mean diameter of the abscess was 11.5 cm. The abscess was located in the right lobe in 21 cases and left lobe in 12. The mean quantity of liquid drained from the abscess was 1060 mL. Outcome was favorable in 28 cases (success rate: 84.8%). In the remaining 5 cases abscess persisted after drain removal including two that required a second drain procedure and three that resolved after medical treatment. The failure rate was 6% (2/33). No patient died. CONCLUSION: Large size and superficial location are risk factors for rupture of liver abscess. Drainage is warranted in patients at risk for this complication. The results of this study show that our "mini-hepatotomy" drainage technique has a high success rate and can achieve good results. PMID- 20025178 TI - [Early outcome of digestive surgery in a tropical setting]. AB - OBJECTIVE: The purpose of this report was to describe early outcome of surgical management of digestive disease in a tropical setting. Study design. This retrospective, descriptive study was carried out in Benin over the three-year period from January 1, 2002 to December 31, 2004. PATIENTS AND METHODS: A total of 613 patients admitted to the intensive care unit (ICU) following surgical treatment for digestive disease were studied. Data were collected on cards from ICU patient admission records, duty register, and patient charts. The data obtained by exact transcription from cards was entered into the Epidata 3.02 software package and analyzed using the Stata 8.0 software package. RESULTS: Patients undergoing surgery for digestive disease accounted for 32% of admissions to the ICU during the study period. Mean patient age was 30 years (range, 1 day to 85 years). Surgery was carried out under emergency conditions in 510 patients and elective conditions in 103. The most common surgical indications were peritonitis, acute bowel occlusion, and malignant tumors. Overall postoperative morbidity was 25.8% with a strong male prevalence (27.6% after emergency procedures and 16.5% after elective procedures). Most complications (74.7%) occurred within 4 days after the procedure. Complications rare occurred after the 6th post-operative day. Overall mortality was 13% (13.3% after emergency procedures and 11.6% after elective surgery). The most frequent cause of death was sepsis. In the vast majority of the cases (78.7%) death occurred in the first 72 hours. CONCLUSION: Post-operative morbidity and mortality remain high in our ICU especially after surgical management of digestive disease. Although this finding is correlated with inadequate technical and human resources, it is mainly due to delayed treatment or slow evacuation time with most patients being admitted in extremely critical condition. Elective surgery was usually performed on patients presenting advanced-stage malignancy. Delayed management with subsequent deterioration of the patient's clinical state was frequently due to prior treatment by practitioners of traditional medicine. PMID- 20025179 TI - [Delay in the management of obstetric complications: study in 7 maternity units in Cameroon]. AB - The purpose of this retrospective, descriptive study conducted in 7 third-level maternity units in Cameroon was to determine maternal mortality rates associated with obstetrical complications and correlate these data with competency of health care staff and time-to-care. Consolidated data for the year 2004 were used to calculate various indicators of maternal health. During the study period 16,005 deliveries were performed with 15,322 live births. Obstetrical complications occurred in 2847 cases leading to a total of 112 maternal deaths. Overall maternal mortality was 699 deaths for 100,000 live births. The complication fatality rate was 3.3 % and was correlated with both competency of health-care staff and time-to-care (p < 0.05). In addition delayed for treatment was a determinant factor in maternal mortality since the risk of death increased with longer time-to-care. PMID- 20025180 TI - [Tetanus in a hospital setting in Bamako, Mali]. AB - AIM: The purpose of this study was to describe the epidemiological, clinical, and prognostic profile of tetanus patients managed in the infectious diseases department of the "Hopital du Point G", in Bamako, Mall. PATIENTS AND METHODS: This retrospective descriptive study was conducted from July 2001 to August 2004. The following data were collected: age, sex, infection route, clinical features, delay to hospitalization, and outcome. RESULTS: A total of 54 patients hospitalized for tetanus were included. Men accounted for 68.5% of cases. Ages ranged from 15 to 66 years with a mean of 39 years. The most common professional activity was farming: 27.8%. The route of infection was determined in 87% of cases and involved cuts in 46.3% of cases located on the lower limbs in 42.6%. Generalized paroxystic tetanus with opisthotonos was the most prevalent form accounting for 18.5% of cases. Localized tetanus accounted for 12.9% of cases. Outcome was unfavorable with a death rate of 38.9%. All deaths occurred within the first week of hospitalization. CONCLUSION: Mass vaccination of the population should be a national priority to reduce tetanus-related morbidity and mortality to levels observed in industrialized countries. PMID- 20025181 TI - [Health impact of working conditions at the Ruashi mine in the Democratic Republic of Congo]. AB - OBJECTIVE: The purpose of this study was to evaluate the health impact of working conditions at the Ruashi mine in the Katanga Province in the Democratic Republic of Congo. MATERIALS AND METHODS: A cross-sectional etiological study was conducted in two cohorts including 100 miners and 109 students. Data necessary to allow comparison disease symptoms in the two cohorts were collected using a questionnaire. Odds ratios (OR) and 95% confidence intervals (95% CI) were estimated. A secondary analysis was performed to compare different mine workstations, i.e., diggers (n=61) versus non-diggers (n=39). Two logistical regression models were used to estimate adjusted OR and 95% CI for the association between occupation/workstation and disease symptoms. RESULTS: In comparison with students, miners exhibited higher frequencies for all symptoms even after adjustment. The highest ORs were observed for musculoskeletal disorders, i.e., low back pain (OR=36.5) and upper (OR = 20.7) or lower (OR = 18.3)] extremity pain. They were followed by respiratory disorders (OR = 5.91) and headache (OR = 5.34). CONCLUSION: The high exposure frequencies and OR's observed in this study underline the negative health impact of mine working conditions. Further occupational study will be needed to obtain and compare more data. PMID- 20025182 TI - [Results of tuberculosis control programs in Madagascar from 1996 to 2004]. AB - OBJECTIVE: Madagascar's national tuberculosis control program has been operational since 1991. The purpose of this article is to provide up-to-date information about the results of this program. METHOD: Data from reports sent to the Tuberculosis Control Department between 1996 and 2004 by diagnosis and treatment centers were retrospectively studied. Special focus was placed on new cases of tuberculosis identified by positive smear. RESULTS: During the study period the annual incidence of new cases of tuberculosis confirmed by positive smear increased from 65 to 82 per 100,000 inhabitants. The highest incidence of new cases was observed in the active population. The treatment success rate rose from 64.4% to 70.8% in patients with positive smear tests. The dropout rate decreased from 21% to 16.5%. However discrepancies were observed between the number of cases diagnosed and number of cases treated. CONCLUSIONS: These findings indicate that tuberculosis control improved slowly over the study period. However these data do not allow identification of strategies to improve program performance. This will require detailed review of data taking into account the context in which they were obtained. PMID- 20025183 TI - [Assessment of a rapid diagnostic test for malaria in rural health care facilities in Senegal]. AB - PURPOSE: The aim of the study was to determine the accuracy of a rapid diagnostic test in confirming presumptive malaria diagnosis in a rural zone of Senegal. Thick blood smear was used as the reference technique for comparison. METHOHDOLOGY: Testing was conducted on children between the ages of 1 and 14 years at three health care facilities located in the Niakhar are from August 2006 to June 2007. If malaria was suspected by the nurse based on clinical findings, two thick smears and one rapid diagnostic test (Core Malaria Pf) were performed. Blood slides were stained in Niakhar and read in Dakar. RESULTS: A total of 474 patients were examined. Three-fourths (75%) of these patients were seen during the rainy season. Malaria was suspected in 335 patients (71%). Rapid tests and thick smears were obtained in 330 of these patients with positive results in 194 (59%) and 180 (55%) respectively. The sensitivity, specificity, positive predictive value and negative predictive value of the rapid test were 96%, 87%, 90% and 95% respectively. CONCLUSION: Our data show that the rapid diagnostic test used in this study exhibits good sensitivity and positive predictive value. Despite its cost this test could be helpful in confirming malaria diagnosis in outlying health care facilities without the necessary resources to perform blood smears. Confirmation is necessary to avoid unwarranted prescription of malaria treatment due to inaccurate clinical diagnosis PMID- 20025184 TI - [A case of meningo-encephalitis due to West Nile virus in Libreville, Gabon]. AB - Acute forms of meningo-encephalitis due to West Nile virus are rare. The purpose of this report is to describe the first case of acute meningoencephalitis due to West Nile virus diagnosed in Gabon. The patient was a 20-year-old Gabonese man. Diagnostic was confirmed by molecular biology. The patient died 12 days after admission to the hospital. This case is discussed based on a review of the literature. PMID- 20025185 TI - [Dermatomyositis and pregnancy: a case in Gabon]. AB - The purpose of this report is to describe a case of successful pregnancy involving a 30-year-old Afican woman presenting dermatomyositis, without use of immunosuppressive treatment. The child was delivered prematurely by caesarean section at 32 weeks of gestation. PMID- 20025186 TI - [Nocardia aroensis and lung infection: first case report in Africa ]. AB - Nocardiosis is a rare disease that is difficult to diagnose. Pulmonary forms are most common in association with a variety of nonspecific symptoms. Up to now isolation of the offending species, i.e., Nocardia aroensis, has been reported only once during the first description in Japan. The purpose of this article is to report the second world case of isolation of the Nocardia aroensis in a 50 year-old immunocompetent African woman. PMID- 20025187 TI - [Nodular liver lesions with fever in a Moroccan man: hepatic brucelloma]. AB - Hepatic brucelloma is an exceptional localization in brucellosis. The purpose of this report is to describe an uncommon case involving a 42-year-old man hospitalized for febrile hepatic cytolysis. Diagnosis was established based on positive rose bengal test results, positive blood culture, hypoechogenic masses on ultrasound, and hypodense enhancing masses on CT scan. Hepatic brucelloma is a focal suppurative lesion occurring after undetected acute brucellosis or undertreated brucellosis. Diagnosis is based on the association of imaging showing characteristic features (hepatic calcifications) and on positive blood culture and serology. First-line treatment should consist of doxycyclin and rifampicin for 2 to 12 months. If medical treatment fails, surgical drainage should be performed. PMID- 20025188 TI - [Athetosis in relation with intracranial tuberculomas in a patient from Burkina Faso]. AB - Athetosis is generally characterized by involuntary movements due to damage of the extrapyramidal tract secondary to neonatal cerebral anoxia or nuclear icterus. The purpose of this report is to describe the case of a 41-year-old man who was admitted to the neurology department of the Ouagadougou teaching hospital in Burkina Faso for right hemiathetosis in relation with intracranial tuberculomas ongoing for two years. Diagnosis was based on clinical findings, i.e., lymph node tuberculosis and positive HIV1 serology; on CT scans showing multiple low density nodular lesions of variable size with annular contrast at the level of the right cerebellum and calcification at the left parietal level and in projection of the left capsulolenticular area; and on favorable response to tuberculosis treatment. PMID- 20025189 TI - [A misleading case of hypereosinophilia revealing colonic adenocarcinoma in a patient from a tropical area]. AB - Although paraneoplastic hypereosinophilia has been documented in patients with colon cancer, this association is rare. The purpose of this report is to describe a case of paraneoplastic hypereosinophilia associated with colic adenocarcinoma. This case underlines the value of methodical investigation of hypereosinophilia in tropical areas where parasitic aetiologies are frequent. PMID- 20025190 TI - [Impact of access to antiretroviral therapy in Cote d'Ivoire]. AB - In 1998 UNAIDS implemented the national drug access initiative (DAI) in Cote d'Ivoire. The Ivorian government took the DAI over in 2000 with the support of the Global Fund and Presidential Emergency Program For AIDS Relief (PEPFAR). The ensuing affordability of antiretroviral therapy (ART), medical staff training, and healthcare equipment allowed Ministry of Health to improve HIV care throughout the country. Since 2008 ART and follow-up monitoring have been free of charge for people living with HIV/AIDS (PLWHA). In January 2009 a total of 57,833 PLWHA received ART and follow-up at 274 HIV care centers. Use of ART has improved the life expectancy of PLWHA. However morbidity and mortality remained high during the first year of ART implementation with respective frequencies of 5-10% person-year (PY) and 2-3% PY. Morbidity was mainly related to infectious disease (tuberculosis and bacteriaemia) and earlier onset of adverse events (AE). In most cases ART has been well tolerated. The main adverse effects have been anemia, neuropathy, skin toxicity and liver enzyme elevation. The incidence of stage 3/4 AE has been low (< 2 %PY). Although overall compliance has been good (<80%), data among children and adults suggest the need for further work to reinforce support mechanisms. Convincing results have been obtained in the management of PLWHA. Nevertheless greater funding and commitment must be given to management of opportunistic infections and side effects and to development of nutrition support services. PMID- 20025191 TI - [Intermittent claudication: sign of epidemiological transition in Gabon]. AB - Africa is in the midst of an epidemiological transition characterized by an increase in degenerative illnesses, in particular cardiovascular disease, and a decrease in infectious illnesses. In this context it is of critical importance to develop simple and effective low-cost tools to identify subjects at high risk for cardiovascular events. Screening for subclavian artery stenosis (SAS) could provide a useful risk indicator. The prevalence of SAS in the general population over the age of 40 years in Gabon is 5.1 %, which is close to prevalence values observed in industrialized countries. Presence of SAS has been associated with hypertension (OR: 5.79, 95% CI: 2.21-15.2; p < 0.05) and male gender (OR 2.34, 95% CI: 1.13-4.83; p < 0.05). These data confirm the epidemiological shift towards cardiovascular diseases and suggest that screening for SAS could be useful to identify subjects at high risk for cardiovascular events who could benefit from preventive strategies. PMID- 20025192 TI - [Temporary tatooing: black henna or harkous?]. AB - Contact sensitization to natural henna (Lawsonia inermis) long used as a skin dye in powder or paste form has rarely, if ever, been observed. Recently a number of anaphylactoid reactions to PPD contained in paints used for temporary tattoos have been described. The purpose of this article is to present 8 cases involving 4 children and 4 adults who developed contact dermatitis after temporary tattoo using "harkous" that contains no henna. The authors review the literature about temporary tattooing. "Harkous" is a mixture of nut gall and clove to which PPD is sometimes added. Many reports have described cases of contact dermatitis due to "black henna". Most of these cases have involved tourists returning from trips. PMID- 20025193 TI - Next generation syndromic surveillance: molecular epidemiology, electronic health records and the pandemic Influenza A (H1N1) virus. AB - In the early phase of the 2009 A (H1N1) pandemic a marked increase in severity and a shift in the age distribution toward younger persons was found, with higher severity reported in patients with pre-existing medical conditions and pregnant women. Consistent with previous pandemics, the age and clinical history of the patients play a critical role in the morbidity and mortality associated with the pandemic virus. This is the first influenza pandemic in the information era, where enormous amounts of information will be available from the pathogen and the patient. Recent advances in molecular techniques have provided an enormous amount of information about pathogens in near real time and at relatively low cost. Electronic Health Records (EHRs) provide another enormously rich set of information about patients, which include patient preconditions, previous exposures, immunization history, presenting complaints, duration and severity of illness, treatment history, and geographic location. An infectious disease is a complex interplay between host and pathogen. The morbidity and mortality of a virus depend on the virus, the patient, and the environment. To evaluate and understand the severity of the pandemic virus and to identify the populations at risk of mild or severe, life-threatening illness, it is compulsory to integrate viral and patient information in a fast and accurate way. Both advances in biomedical informatics with the creation of EHRs and molecular techniques provide the framework to achieve these aims. PMID- 20025194 TI - Evolutionary dynamics of N-glycosylation sites of influenza virus hemagglutinin. AB - The hemagglutinin protein of influenza virus bears several sites of N-linked asparagine glycosylation. The number and location of these sites varies with strain and substrain. The human H3 hemagglutinin has gained several glycosylation sites on the antigenically important globular head since its introduction to humans, presumably due to selection. Although there is abundant evidence that glycosylation can affect antigenic and functional properties of the protein, direct evidence for selection is lacking. We have analyzed gain and loss of glycosylation sites on the side branches of a large phylogenetic tree of H(3) HA1 sequences (branches off of the main, long-term line of descent). Side branches contrast with the main line of descent: losses of glycosylation sites are not uncommon, and they outnumber gains. Although other explanations are possible, this observation is consistent with weak selection for glycosylation sites or a more complicated pattern of selection. Furthermore, terminal and internal branches differ with respect to rates of gain and loss of glycosylation sites. This pattern would not be expected under selective neutrality, but is easily explained by weak selection or selection that changes with the immune state of the host population. Thus, it provides evidence that selection acts on the glycosylation state of hemagglutinin. PMID- 20025195 TI - The early molecular epidemiology of the swine-origin A/H1N1 human influenza pandemic. AB - Swine-origin pandemic human influenza A virus (H1N1pdm) has spread rapidly around the world since its initial documentation in April 2009. Here we have updated initial estimates of the rate of molecular evolution and estimates of the time of origin of this virus in the human population using the large number of viral sequences made available as part of the public health response to this global pandemic. Currently sampled H1N1pdm sequences share a most recent common ancestor in the first 7 weeks of 2009 with the implication that the virus was transmitting cryptically for up to 3 months prior to recognition. A phylogenetic reconstruction of the data shows that the virus has been circling the globe extensively with multiple introductions into most geographical areas. PMID- 20025196 TI - Adaptive vaccination strategies to mitigate pandemic influenza: Mexico as a case study. AB - In this modeling work, we explore the effectiveness of various age-targeted vaccination strategies to mitigate hospitalization and mortality from pandemic influenza, assuming limited vaccine supplies. We propose a novel adaptive vaccination strategy in which vaccination is initiated during the outbreak and priority groups are identified based on real-time epidemiological data monitoring age-specific risk of hospitalization and death. We apply this strategy to detailed epidemiological and demographic data collected during the recent swine A/H1N1 outbreak in Mexico. We show that the adaptive strategy targeting age groups 6-59 years is the most effective in reducing hospitalizations and deaths, as compared with a more traditional strategy used in the control of seasonal influenza and targeting children under 5 and seniors over 65. Results are robust to a number of assumptions and could provide guidance to many nations facing a recrudescence of A/H1N1v pandemic activity in the fall and likely vaccine shortages. PMID- 20025197 TI - Transmission of influenza virus in temperate zones is predominantly by aerosol, in the tropics by contact: a hypothesis. AB - Using the guinea pig model, we have previously shown that the aerosol transmission of a seasonal human influenza virus is blocked by humid (80% relative humidity) or warm (30 degrees C) ambient conditions. In contrast, we found that transmission by a contact route proceeded at high efficiency despite increased temperature or humidity. Based on these findings, and the observed seasonal behavior of influenza viruses in various regions of the world, we hypothesize herein that the predominant mode of influenza virus transmission differs in temperate and tropical climates. Specifically, we predict that aerosol transmission predominates during the winter season in temperate regions, while contact is the major mode of spread in the tropics. With this idea in mind, possible explanations for the current summer-time spread of swine-origin influenza viruses are discussed. PMID- 20025198 TI - Swine origin influenza A (H1N1) virus and ICU capacity in the US: are we prepared? AB - We developed a model simulating the potential impact of influenza H1N1 pandemic on the volume of acute respiratory failure requiring mechanical ventilation (ARF MV) and the accompanying mortality rate in the US. We calculate that 46 million people will contract the infection, resulting in 2.7 million hospitalizations, 331,587 episodes of ARF-MV and nearly 200,000 deaths, suggesting that the US may require the ability to provide MV at a volume approximately 33% over the current annual use. PMID- 20025199 TI - Spatiotemporal dynamics in the early stages of the 2009 A/H1N1 influenza pandemic. AB - Epidemiology and public health planning will increasingly rely on the analysis of genetic sequence data. The ongoing influenza A/H1N1 pandemic may represent a tipping point in this trend, with A/H1N1 being the first human pathogen routinely genotyped from the beginning of its spread. To take full advantage of this genetic information, we introduce a novel method to reconstruct the spatiotemporal dynamics of outbreaks from sequence data. The approach is based on a new paradigm were ancestries are inferred directly rather than through the reconstruction of most recent common ancestors (MRCAs) as in phylogenetics. Using 279 A/H1N1 hemagglutinin (HA) sequences, we confirm the emergence of the 2009 flu pandemic in Mexico. The virus initially spread to the US, and then to the rest of the world with both Mexico and the US acting as the main sources. While compatible with current epidemiological understanding of the 2009 H1N1 pandemic, our results provide a much finer picture of the spatiotemporal dynamics. The results also highlight how much additional epidemiological information can be gathered from genetic monitoring of a disease outbreak. PMID- 20025200 TI - Social versus independent interest in 'bird flu' and 'swine flu'. AB - The explosion of interest in H1N1, more popularly called 'swine flu', across the world, from late April to early May 2009, exemplified how information transmission in modern online society can affect the spread of the disease itself. A simple but effective model based on cultural evolutionary theory can characterise in such data the effective degree of social transmission versus independent decision. In a novel approach that applies this model to Google Trends search data, we find significant differences in social transmission of the exact phrase 'swine flu' in 2009, compared with 'bird flu' in 2005. The methodology can thus inform policies for addressing public awareness of health issues, which can be more effective with knowledge of how the information is being spread or learned. PMID- 20025201 TI - Behavioural responses to influenza pandemics: what do we know? AB - The emergence of the novel A/H1N1 virus has made pandemic preparedness a crucial issue for public health worldwide. Although the epidemiological aspects of the three 20th century influenza pandemics have been widely investigated, little is known about population behaviour in a pandemic situation. Such knowledge is however critical, notably for predicting population compliance with non pharmaceutical interventions. This paper reviews the relevant scientific literature for the 1918-1920, 1957-1958, 1969-1969 influenza epidemics and the 2003 SARS outbreak. Although the evidence base of most non pharmaceutical interventions (NPIs) and personal protection measures is debated, it appears on the basis of past experience that NPIs implemented the most systematically, the earliest, and for the longest time could reduce overall mortality rates and spread out epidemic peaks. Adequate, transparent, and targeted communication on the part of public health authorities would be also of crucial importance in the event of a serious influenza pandemic. PMID- 20025202 TI - Point of care strategy for rapid diagnosis of novel A/H1N1 influenza virus. AB - In late June 2009, we implemented for public hospitals of Marseille Point Of Care strategy for rapid diagnosis of novel A/H1N1 influenza virus. During two months, we have tested more than 900 specimens in both Point Of Care laboratories. We believe that implementation of Point of Care strategy for the largest number of suspects cases may improve quality of patients care and our knowledge of the epidemiology of the pandemic. PMID- 20025203 TI - SeqMonitor: influenza analysis pipeline and visualization. AB - Unprecedented sequencing effort has led to daily submissions of influenza genomes to public repositories such as the NCBI GenBank. With the decreasing cost of genome sequencing, it is expected that rapidly evolving viruses such as influenza will be sampled in even greater depth in the future. Keeping analyses up to date and managing this data is a prime concern for researchers and public-health officials alike. We have developed an influenza sequence pipeline, polymorphism data warehouse, and an interactive web-based analysis program to assist in managing the flow of sequence data. The system provides a framework for studying polymorphic associations with various metadata, for downloading subsets based on metadata criteria, as well as for tracking polymorphisms geographically and temporally. SeqMonitor is accessible at http://ratite.cs.dal.ca/SeqMonitor. PMID- 20025204 TI - Pandemic influenza dynamics and the breakdown of herd immunity. AB - Few if any attempts have been made to derive forecasts for the ongoing H1N1 pandemic as extrapolated from knowledge of seasonal influenza. Even simple back of-the-envelope calculations are lacking. In this note we use first-approximation parameter estimates for the SIR model to compare seasonal and pandemic influenza, and then explore the implications of the existing classical epidemiological theory. In particular, we note the dramatic nonlinear increase in attack rate as a function of the percentage of susceptibles initially present in the population. This has severe consequences for the pandemic, given the general lack of immunity in the global population. PMID- 20025205 TI - Economic cost and health care workforce effects of school closures in the U.S. AB - School closure is an important component of U.S. pandemic flu mitigation strategy, but has important costs. We give estimates of both the direct economic and health care impacts for school closure durations of 2, 4, 6, and 12 weeks under a range of assumptions. We find that closing all schools in the U.S. for four weeks could cost between $10 and $47 billion dollars (0.1-0.3% of GDP) and lead to a reduction of 6% to 19% in key health care personnel. PMID- 20025206 TI - Green tea catechins inhibit the endonuclease activity of influenza A virus RNA polymerase. AB - The influenza A RNA polymerase possesses endonuclease activity to digest the host mRNA. Thus this endonuclease domain can be a target of anti-influenza A virus drug. Here we report that green tea catechins inhibit this viral endonuclease activity and that their galloyl group is important for their function. Docking simulations revealed that catechins with galloyl group fit well into the active pocket of the endonuclease domain to enable stable binding. Our results provide useful data that make it possible to refine and optimize catechin-based drug design more readily for stability. PMID- 20025208 TI - Langmuir-Schaeffer films from a pi-pi stacking perylenediimide dye: organization and charge transfer properties. AB - The organization of pi-pi stacking perylenediimide (PDI) derivative, PDI12, was studied in solution and in thin films. Films were prepared with the Langmuir Schaeffer (LS) method and characterized by means of AFM, optical profilometry, steady-state absorption, emission, fluorescence lifetime, and transient photovoltage measurements. The columnar aggregates observed previously in PDI12 solutions and in spin-coated films persist also in LS films. Because of the specific conditions during the preparation of the LS film, i.e., hydrophobic interactions and lateral compression, the columnar aggregates seem to organize with their long axis perpendicular to the layer plane whereas in spin-coated films the columns were oriented parallel to the layer plane. According to AFM and profilometer results, the thickness of LS monolayer of PDI12 is 10 nm, indicating that it consists mainly of aggregates, each containing approximately 30 monomers. Intermolecular photoinduced energy and electron transfer processes in C(60)|PDI12 double layer junction were studied. The fluorescence lifetime of PDI12 film is exceptionally long, but the quenching is very efficient in the presence of C(60). In charge transfer studies, long-lived photovoltage signal was observed for the double layer. Results of this work indicate that PDI12 acts as an electron acceptor and fullerene C(60) as an electron donor. PMID- 20025209 TI - Solid-solid phase transition of n-alkanes in multiple nanoscale confinement. AB - The crystallization behavior of n-C(19)H(40)/SiO(2) nanosphere composites was investigated by a combination of differential scanning calorimetry (DSC) and temperature-dependent X-ray diffraction (XRD). Three kinds of confined alkanes with different solid-solid phase transition supercoolings and a surface (or interface) freezing monolayer of n-C(19)H(40) at the bulk liquid/SiO(2) interface were found in the composites at high SiO(2) loading. The surface freezing monolayer induces the chain packing of bulk alkanes by forming a 2D close-packed arrangement without long-range positional ordering in 3D space. A homogeneous nucleation and growth mechanism is found for the solid-solid transition in confined geometry, in which the supercooling of the transition is sensitive to the confined size. PMID- 20025210 TI - Long-range assembly of DNA into nanofibers and highly ordered networks using a block copolymer approach. AB - A simple method to introduce the long-range order achieved by block copolymers into DNA structures is described. This results in the hierarchical assembly of short DNA strands into a new one-dimensional material, with high aspect ratio and the ability to further align into highly ordered surfaces over tens of micrometers. Fibers derived from biological materials have a wide range of potential applications, such as scaffolds for nanowires and one-dimensional (1D) materials, templates for tissue growth, and ligand display tools for multivalent biological interactions. Fibers derived from short DNA strands are an attractive class of materials, as they combine long-range 1D ordering with the programmability of DNA, and its ability to undergo structure switching with specifically added DNA strands. Here, we present the first examples of long fibers self-assembled from short (10-20 base-pairs), blunt-ended DNA strands. This was accomplished by covalently attaching a dendritic oligoethylene glycol (OEG) unit to a DNA strand to form a dendritic DNA molecule (D-DNA). Hybridization of this unit with complementary DNA creates a block copolymer/double-stranded DNA architecture, which readily undergoes self-assembly into long fibers upon the addition of a selective solvent. These fibers can further align into parallel rows, to yield highly ordered micrometer-sized surfaces. We demonstrate that a DNA nanotechnology motif, a three-helix DNA bundle, can also be readily induced to form long fibers upon incorporation of D DNA. Thus, this provides a straightforward method to introduce hierarchical long range ordering into DNA motifs, simply through hybridization with short D-DNA strands. PMID- 20025211 TI - Upconversion for white light generation by a single compound. AB - A new multiphoton upconversion process for rare earth oxide powders is reported, using infrared laser diodes. In vacuum, multiphoton excitation produces intense upconversion emission in the visible spectral range, and Tm(2)O(3) or Yb(2)O(3) achieves pure white upconversion using 808 or 975 nm excitation, respectively. PMID- 20025212 TI - Nonacidic inhibitors of human microsomal prostaglandin synthase 1 (mPGES 1) identified by a multistep virtual screening protocol. AB - Microsomal prostaglandin E(2)-synthase (mPGES-1) is a target for future anti inflammatory drugs. Inhibitors of mPGES-1 mimicking prostaglandin E(2) often interact with cyclooxygenases (COXs) 1 and 2, leading to unwanted side effects. Selective inhibitors of mPGES-1 can be obtained by deliberate abdication of the acidic groups, which are an important feature of COX inhibition. Here, we present a successful virtual screening study that results in a potent nonacidic mPGES-1 inhibitor lacking COX inhibition. PMID- 20025213 TI - Near-infrared reflectance spectroscopy predicts protein, starch, and seed weight in intact seeds of common bean ( Phaseolus vulgaris L.). AB - The objective of this study was to explore the potential of near-infrared reflectance (NIR) spectroscopy to determine individual seed composition in common bean ( Phaseolus vulgaris L.). NIR spectra and analytical measurements of seed weight, protein, and starch were collected from 267 individual bean seeds representing 91 diverse genotypes. Partial least-squares (PLS) regression models were developed with 61 bean accessions randomly assigned to a calibration data set and 30 accessions assigned to an external validation set. Protein gave the most accurate PLS regression, with the external validation set having a standard error of prediction (SEP) = 1.6%. PLS regressions for seed weight and starch had sufficient accuracy for seed sorting applications, with SEP = 41.2 mg and 4.9%, respectively. Seed color had a clear effect on the NIR spectra, with black beans having a distinct spectral type. Seed coat color did not impact the accuracy of PLS predictions. This research demonstrates that NIR is a promising technique for simultaneous sorting of multiple seed traits in single bean seeds with no sample preparation. PMID- 20025214 TI - Potential-dependent studies on the interaction between phenylalanine-substituted bombesin fragments and roughened Ag, Au, and Cu electrode surfaces. AB - In this work, we report systematic surface-enhanced Raman spectroscopy (SERS) and generalized two-dimensional correlation analysis (G2DCA) studies of the structures of five specifically modified phenylalanine-substituted C-terminal bombesin 6-14 fragments (BN(6-14)). The fragments studied have all been tested as chemotherapeutic agents in cancer therapy, and they form amino acid sequences in bombesin: cyclo[d-Phe(6),His(7),Leu(14)]BN(6-14), [D-Phe(6),Leu-NHEt(13),des Met(14)]BN(6-14), [D-Phe(6),Leu(13)-((R))-p-Cl-Phe(14)]BN(6-14), [D-Phe(6),beta Ala(11),Phe(13),Nle(14)]BN(6-14), and [D-Tyr(6),beta-Ala(11),Phe(13),Nle(14)]BN(6 14). We adsorbed these fragments onto roughened Ag, Au, and Cu electrode surfaces, using a potential range from -1.200 to 0.400 V, at physiological pH. We compared the adsorption mechanism of each fragment on these substrates, as well any changes observed with varying electrode potential, to determine the relationship between adsorption strength and geometry of each of the peptides wherever it was possible. For example, we showed that none of these fragments directly interact with the Ag, Au, and Cu surfaces via residues of Phe (phenylalanine) and Trp(8) (L-tryptophane at position 8 of the BN amino acid sequence) or by an amide bond, due to a very small shift in wavenumber of their characteristic vibrations. Specific interactions were recognized from the broadening, wavenumber shift, and increase in intensity of the W18 Trp(8) mode near 759 cm(-1) and decrease in nu(12) vibration frequency of the Phe residue. In general, more intense SERS bands were observed due to the Phe ring, compared with the Trp(8) ring, which suggested a preferential adsorption of phenylalanine over tryptophane. For [D-Tyr(6),beta-Ala(11),Phe(13),Nle(14)]BN(6-14), the data also suggest some interaction of a D-Tyr(6) residue (D-tyrosine at position 6). Finally, only slight rearrangements of these moieties on the substrates are observed with changes in electrode potential. PMID- 20025215 TI - Anti-arthritic effects and toxicity of the essential oils of turmeric (Curcuma longa L.). AB - Turmeric (Curcuma longa L., Zingiberaceae) rhizomes contain two classes of secondary metabolites, curcuminoids and the less well-studied essential oils. Having previously identified potent anti-arthritic effects of the curcuminoids in turmeric extracts in an animal model of rheumatoid arthritis (RA), studies were undertaken to determine whether the turmeric essential oils (TEO) were also joint protective using the same experimental model. Crude or refined TEO extracts dramatically inhibited joint swelling (90-100% inhibition) in female rats with streptococcal cell wall (SCW)-induced arthritis when extracts were administered via intraperitoneal injection to maximize uniform delivery. However, this anti arthritic effect was accompanied by significant morbidity and mortality. Oral administration of a 20-fold higher dose TEO was nontoxic, but only mildly joint protective (20% inhibition). These results do not support the isolated use of TEO for arthritis treatment but, instead, identify potential safety concerns in vertebrates exposed to TEO. PMID- 20025216 TI - Double-stranded supramolecular assembly through salt bridge formation between rigid and flexible amidine and carboxylic acid strands. AB - A series of monomeric strands consisting of m-terphenyl backbones with chiral rigid C-linked (3) and flexible N-linked (5) formamidines and achiral carboxylic acid (4) and flexible carboxymethyl (6) residues were synthesized, and their duplex formations through amidinium-carboxylate salt bridges were investigated by NMR, circular dichroism (CD), and UV-visible spectroscopies. The salt bridge derived duplex formation was largely dependent on the structures of the formamidine and carboxylic acid strands, and the C-linked formamidine strand 3 formed a more stable duplex with the complementary carboxylic acid strands (4 and 6) than did the flexible N-linked formamidine strand 5. The single crystal X-ray analysis revealed that the duplex 5.4 has a skewed right-handed double helical structure. A complementary duplex dimer was also synthesized from the dimers of 5 and 4 joined by diacetylene linkers. Variable-temperature CD measurements indicated that the duplex possesses a dynamic double helical structure resulting from the flexible N-linked formamidine units. PMID- 20025217 TI - Design of natural food antioxidant ingredients through a chemometric approach. AB - In the present work, an environmentally friendly extraction process using subcritical conditions has been tested to obtain potential natural food ingredients from natural sources such as plants, fruits, spirulina, propolis, and tuber, with the scope of substituting synthetic antioxidants, which are subject to regulation restrictions and might be harmful for human health. A full characterization has been undertaken from the chemical and biochemical point of view to be able to understand their mechanism of action. Thus, an analytical method for profiling the compounds responsible for the antioxidant activity has been used, allowing the simultaneous determination of water-soluble vitamins, fat soluble vitamins, phenolic compounds, carotenoids, and chlorophylls in a single run. This information has been integrated and analyzed using a chemometrical approach to correlate the bioactive compounds profile with the antioxidant activity and thus to be able to predict antioxidant activities of complex formulations. As a further step, a simplex centroid mixture design has been tested to find the optimal formulation and to calculate the effect of the interaction among individual extracts in the mixture. PMID- 20025218 TI - Rhodium-catalyzed [3 + 2] annulation of indoles. AB - An effective Rh(2)(S-DOSP)(4)-catalyzed asymmetric cyclopentannulation of indolyl rings has been developed. Depending on the substitution pattern of the indole, two distinct regioisomeric products can be generated. These studies demonstrate that rhodium-catalyzed reactions of donor/acceptor carbenoids proceeding by means of zwitterionic intermediates can be carried out with very high asymmetric induction. PMID- 20025219 TI - Compressing the most hydrogen-rich inorganic ion. AB - Motivated by the potential high-temperature superconductivity of "chemically precompressed" hydrogen-rich compounds, the high pressure phases of the ionic salt BaReH(9) are explored theoretically. We find that the compound adapts to compression not only by structural distortions or increase of coordination number, but also through evolution of discrete H(2) units which fill up interspace gaps. This last structural change is associated with a dramatic lowering of metallization pressure, so that BaReH(9) can be expected to turn metallic at the onset of the H(2)-containing phase (51 GPa). PMID- 20025220 TI - Relationship between grazing lamb growth rate and blood plasma analytes as profiled by gas chromatography with time-of-flight mass spectrometry (GC-TOF/MS). AB - There can be considerable variation in the performance of individual lambs grazing on the same pasture. Gas chromatography with time-of-flight mass spectrometry (GC-TOF/MS) was used to profile the relative abundances of metabolites in plasma from growing lambs to determine any correlation effects between plasma metabolites and liveweight gain. Analysis of relative abundance of 336 analyte clusters and liveweight gain revealed that the growth rates of female lambs were significantly positively correlated with 5 analyte clusters and negatively correlated with 5 other analyte clusters. Growth rates of male lambs were likewise significantly positively correlated with 9 analyte clusters and negatively with 5 analyte clusters. Analytes identified as being associated with lamb growth rate included the amino acids valine, methionine, phenylalanine, cystine and asparagine, and oxalic acid, phenylacetic acid, and phosphoric acid. A number of currently unidentified analytes were significantly correlated with growth rate. Stepwise regression of the analytes on lamb growth rate yielded relationships that accounted for 48% and 58% of the variation in female and male lamb growth rates, respectively. This study demonstrated that by using GC-TOF/MS in combination with multivariate statistical techniques it is possible to correlate the presence of specific analytes in sheep plasma with growth rate. PMID- 20025221 TI - Characterization of 14 raspberry cultivars by solid-phase microextraction and relationship with gray mold susceptibility. AB - Fourteen raspberry varieties were evaluated over two cropping seasons by solid phase microextraction (SPME) followed by gas chromatography-mass spectrometry. Thirty-six compounds were fully identified, and 10 more compounds were tentatively identified. Despite interannual variability, raspberry varieties can be divided in two main groups on the basis of terpenes and C-13 norisoprenoids. Susceptibility toward Botrytis cinerea , one of the most relevant pathogenic fungi for soft fruits during storage, was also evaluated. On the basis of volatile profiles, it was possible to highlight the relationship between different volatile compounds and resistance to B. cinerea. Volatile profiles and Botrytis susceptibility of the different raspberry varieties evaluated should assist future breeding programs. PMID- 20025222 TI - InAs(ZnCdS) quantum dots optimized for biological imaging in the near-infrared. AB - We present the synthesis of InAs quantum dots (QDs) with a ZnCdS shell with bright and stable emission in the near-infrared (NIR, 700-900 nm) region for biological imaging applications. We demonstrate how NIR QDs can image tumor vasculature in vivo at significantly deeper penetration depths and with higher contrast than visible emitting CdSe(CdS) QDs. Targeted cellular labeling is also presented and may enable multiplexed and low autofluorescence cellular imaging. PMID- 20025224 TI - In vitro bioavailability of iron from the heme analogue sodium iron chlorophyllin. AB - The use of heme analogues from vegetable origin could provide an alternative iron source of potentially high bioavailability. Sodium iron chlorophyllin is a water soluble semisynthetic chlorophyll derivative where the magnesium in the porphyrin ring has been substituted by iron. We have used an in vitro model that combines gastric and intestinal digestion followed by intestinal iron uptake in Caco-2 cells to determine the bioavailability of iron from sodium iron chlorophyllin. Our results demonstrate that sodium iron chlorophyllin is stable under simulated gastrointestinal conditions and is able to deliver bioavailable iron to Caco-2 cells. Similar to the heme, the bioavailability of iron from sodium iron chlorophyllin is dependent on the food matrix, and it was inhibited by calcium. Potentially, sodium iron chlorophyllin could be used as an iron fortificant from vegetable origin with high bioavailability. PMID- 20025223 TI - Compact biocompatible quantum dots via RAFT-mediated synthesis of imidazole-based random copolymer ligand. AB - We present a new class of polymeric ligands for quantum dot (QD) water solubilization to yield biocompatible and derivatizable QDs with compact size (approximately 10-12 nm diameter), high quantum yields (>50%), excellent stability across a large pH range (pH 5-10.5), and low nonspecific binding. To address the fundamental problem of thiol instability in traditional ligand exchange systems, the polymers here employ a stable multidentate imidazole binding motif to the QD surface. The polymers are synthesized via reversible addition-fragmentation chain transfer-mediated polymerization to produce molecular weight controlled monodisperse random copolymers from three types of monomers that feature imidazole groups for QD binding, polyethylene glycol (PEG) groups for water solubilization, and either primary amines or biotin groups for derivatization. The polymer architecture can be tuned by the monomer ratios to yield aqueous QDs with targeted surface functionalities. By incorporating amino PEG monomers, we demonstrate covalent conjugation of a dye to form a highly efficient QD-dye energy transfer pair as well as covalent conjugation to streptavidin for high-affinity single molecule imaging of biotinylated receptors on live cells with minimal nonspecific binding. The small size and low serum binding of these polymer-coated QDs also allow us to demonstrate their utility for in vivo imaging of the tumor microenvironment in live mice. PMID- 20025225 TI - Enantiomeric analysis of chiral compounds in irradiated foods using multidimensional gas chromatography. AB - The usefulness of both solid phase microextraction (SPME) and multidimensional gas chromatography (MDGC) coupled to mass spectrometry (MS) to detect chiral compounds in irradiated cheese was evaluated. The enantiomeric resolution of relevant chiral aroma compound was achieved by analyzing the extracts obtained from SPME by means of a permethylated beta-CD stationary phase as the main column of the multidimensional system to separate specific selected cuts containing components unresolved in the first dimension. The proposed procedure allowed to determine, in less than 90 min, that no significant variations had been produced in the stereoisomeric distribution of limonene, 3-hydroxybutanone, and 2,3 butanediol in cheese when applying irradiation doses ranging from 0 to 8 kGy. PMID- 20025226 TI - Small-angle X-ray scattering study of the effect of pH and salts on 11S soy glycinin in the freeze-dried powder and solution states. AB - The nanostructures from powders of native protein, glycinin, and corresponding solutions from which the powders have been formed, have been studied as a function of pH and 1 M salts using small-angle X-ray scattering. All powders showed Porod scattering with the exception of that prepared from the solution close to pI which displayed fractal behavior. Well-defined Bragg peaks in the powder scattering at pH 5, pH 7, and 1 M NaCl indicate the presence of long-range order. The scattering from solutions at pH 7, pH 9, and 1 M NaCl can be described well on the basis of particles derived from the known atomic structures of homohexameric glycinin. Extreme acidic (pH 2) and basic (pH 11) environments lead to the partial denaturation of glycinin. Decreasing the pH to 2 initiates dissociation of the hexameric structure, while increasing the pH to 11, as well as the presence of 1 M NaSCN, results in the formation of large unimodal particles. This is reflected by "featureless" SAXS patterns for both powders and solutions. PMID- 20025227 TI - Anharmonicities and isotopic effects in the vibrational spectra of X-.H2O, .HDO, and .D2O [X = Cl, Br, and I] binary complexes. AB - Vibrational predissociation spectra of the argon-tagged halide monohydrates, X(-) .H(2)O.Ar (X = Cl, Br, or I), are recorded from approximately 800 to 3800 cm(-1) by monitoring the loss of the argon atom. We use this set of spectra to investigate how the spectral signatures of the hydrogen-bonding and large amplitude hindered rotations of the water molecule are affected by incremental substitution of the hydrogen atoms by deuterium. All six vibrational modes of the X(-).H(2)O complexes are assigned through fundamental transitions, overtones, or combination bands. To complement the experimental study, harmonic and reduced dimensional calculations of the vibrational spectra are performed based on the MP2/aug-cc-pVTZ level of theory and basis set. Comparison of these results with those from the converged six-dimensional calculations of Rheinecker and Bowman [J. Chem. Phys. 2006, 125, 133206.] show good agreement, with differences smaller than 30 cm(-1). The simpler method has the advantage that it can be readily extended to the heavier halides and was found to accurately recover the wide range of behaviors displayed by this series, including the onset of tunneling between equivalent minima arising from the asymmetrical (single ionic hydrogen bonded) equilibrium structures of the complexes. PMID- 20025228 TI - Spider-mediated flux of PCBs from contaminated sediments to terrestrial ecosystems and potential risks to arachnivorous birds. AB - We investigated aquatic insect utilization and PCB exposure in riparian spiders at the Lake Hartwell Superfund site (Clemson, SC). We sampled sediments, adult chironomids, terrestrial insects, riparian spiders (Tetragnathidae, Araneidae, and Mecynogea lemniscata), and upland spiders (Araneidae) along a sediment contamination gradient. Stable isotopes (delta(13)C, delta(15)N) indicated that riparian spiders primarily consumed aquatic insects whereas upland spiders consumed terrestrial insects. PCBs in chironomids (mean 1240 ng/g among sites) were 2 orders of magnitude higher than terrestrial insects (15.2 ng/g), similar to differences between riparian (820-2012 ng/g) and upland spiders (30 ng/g). Riparian spider PCBs were positively correlated with sediment concentrations for all taxa (r(2) = 0.44-0.87). We calculated spider-based wildlife values (WVs, the minimum spider PCB concentrations causing physiologically significant doses in consumers) to assess exposure risks for arachnivorous birds. Spider concentrations exceeded WVs for most birds at heavily contaminated sites and were approximately 14-fold higher for the most sensitive species (chickadee nestlings, Poecile spp.). Spiders are abundant and ubiquitous in riparian habitats, where they depend on aquatic insect prey. These traits, along with the high degree of spatial correlation between spider and sediment concentrations we observed, suggest that they are model indicator species for monitoring contaminated sediment sites and assessing risks associated with contaminant flux into terrestrial ecosystems. PMID- 20025230 TI - Real-time monitoring of fragrance release from cotton towels by low thermal mass gas chromatography using a longitudinally modulating cryogenic system for headspace sampling and injection. AB - An innovative headspace sampling and injection system for gas chromatography was designed using a longitudinally modulating cryogenic system mounted around the sampling loop of a two-position loop injector. The setup was hyphenated to a fast low thermal mass gas chromatograph, allowing transient concentrations of semivolatile analytes to be monitored in real time with a time resolution of 4.5 min. The performance of the instrument, and in particular its cryotrapping efficiency, was characterized using a mixture of long-chain alkanes, methyl esters, ethyl esters, and alcohols of different volatilities. The device was found to be ideally suited to the analysis of semivolatile compounds with boiling points ranging between 190 and 320 degrees C, which are typical for a majority of perfumery raw materials. The new instrument was successfully used to monitor the release of eight odorant compounds from cotton towels to which fabric softener had been applied that alternatively contained the fragrance in free form or in microencapsulated form. The analytical results, unprecedented in their level of precision and time resolution for such an application, evidenced the major impact of microencapsulation technology on the kinetics of fragrance release during the drying of the towels and on the triggering of additional fragrance release by applying mechanical stress to the fabric to rupture the microcapsule walls. PMID- 20025229 TI - Alaskan wild berry resources and human health under the cloud of climate change. AB - Wild berries are integral dietary components for Alaska Native people and a rich source of polyphenolic metabolites that can ameliorate metabolic disorders such as obesity and diabetes. In this study, five species of wild Alaskan berries (Vaccinium ovalifolium , Vaccinium uliginosum , Rubus chamaemorus , Rubus spectabilis , and Empetrum nigrum) were screened for bioactivity through a community-participatory research method involving three geographically distinct tribal communities. Compositional analysis by HPLC and LC-MS(2) revealed substantial site-specific variation in anthocyanins (0.01-4.39 mg/g of FW) and proanthocyanidins (0.74-6.25 mg/g of FW) and identified A-type proanthocyanidin polymers. R. spectabilis increased expression levels of preadipocyte factor 1 (182%), and proanthocyanidin-enriched fractions from other species reduced lipid accumulation in 3T3-L1 adipocytes. Selected extracts reduced serum glucose levels in C57BL/6J mice by up to 45%. Local observations provided robust insights into effects of climatic fluctuations on berry abundance and quality, and preliminary site-specific compositional and bioactivity differences were noted, suggesting the need to monitor this Alaska Native resource as climate shifts affect the region. PMID- 20025231 TI - Vibrational analysis of amino acids and short peptides in hydrated media. VI. Amino acids with positively charged side chains: L-lysine and L-arginine. AB - In two recent reports of the same series (J. Phys. Chem. B 2007, 111, 1470-1477 and J. Phys. Chem. B 2009, 113, 3169-3178), we have described the geometrical and vibrational analysis of glycine and amino acids (AAs) with hydrophobic side chains through the joint use of optical spectroscopy and quantum mechanical calculations. Here, we report Raman scattering and Fourier-Transform Infrared (FT IR) Attenuated Total Reflectance (ATR) spectra measured from the aqueous solutions (H(2)O and D(2)O) of L-lysine and L-arginine, i.e. two alpha-AAs with positively charged hydrophilic side chains. The discussion on the vibrational features of both AAs could be carried out thanks to the theoretical calculations performed by means of the Density Functional Theory (DFT) approach at the B3LYP/6 31++G* level. We have analyzed the influence of implicit (with a polarizable dielectric continuum) and explicit (by means of an H(2)O cluster interacting with H-donor and H-acceptor sites of AAs) hydration models. In addition, through the calculated geometrical parameters and vibrational wavenumbers, a discussion was performed on the effect of the Cl(-) anion interacting with the positively charged side chains of explicitly hydrated AAs. PMID- 20025232 TI - Electrochromic enhancement of latent fingerprints on stainless steel surfaces. AB - The visualization of latent fingerprints on a metallic (stainless steel) surface is described by means of spatially selective deposition of an electrochromic polymer (polyaniline). Inhibition of electrochemical processes on areas of the surface masked by the fingerprint results in polymer deposition generating a negative image of the fingermark. By variation of the applied potential, the polymer optical characteristics can be continuously and reversibly adjusted to optimize visual contrast of the fingerprint. PMID- 20025233 TI - Silica gel-catalyzed one-pot syntheses in water and fluorescence properties studies of 5-amino-2-aryl-3H-chromeno[4,3,2-de][1,6]naphthyridine-4-carbonitriles and 5-amino-2-aryl-3H-quinolino[4,3,2-de][1,6]naphthyridine-4-carbonitriles. AB - The silica gel-catalyzed synthesis of 5-amino-2-aryl-3H-chromeno[4,3,2 de][1,6]naphthyridine-4-carbonitriles and 5-amino-2-aryl-3H-quinolino[4,3,2 de][1,6]naphthyridine-4-carbonitriles were simply achieved upon the one-pot cascade reaction of malononitrile with substituted 2-hydroxyacetophenone (or 2 aminoacetophenone) and aromatic aldehyde in aqueous media. The mechanistic investigation results based on electrospray ionization mass spectrometry (ESI-MS) indicated that malononitrile displayed a dual role during this transformation. Thirteen bonds were cleaved and 12 new bonds were constructed in the formation of 5-amino-2-aryl-3H-chromeno[4,3,2-de][1,6]naphthyridine-4-carbonitriles, while only 2 H(2)O molecules were removed. The fluorescence properties screening showed five new compounds have high fluorescence quantum yields. PMID- 20025234 TI - High mobility, printable, and solution-processed graphene electronics. AB - The ability to print graphene sheets onto large scale, flexible substrates holds promise for large scale, transparent electronics on flexible substrates. Solution processable graphene sheets derived from graphite can form stable dispersions in solutions and are amenable to bulk scale processing and ink jet printing. However, the electrical conductivity and carrier mobilities of this material are usually reported to be orders of magnitude poorer than that of the mechanically cleaved counterpart due to its higher density of defects, which restricts its use in electronics. Here, we show that by optimizing several key factors in processing, we are able to fabricate high mobility graphene films derived from large sized graphene oxide sheets, which paves the way for all-carbon post-CMOS electronics. All-carbon source-drain channel electronics fabricated from such films exhibit significantly improved transport characteristics, with carrier mobilities of 365 cm(2)/(V.s) for hole and 281 cm(2)/(V.s) for electron, measured in air at room temperature. In particular, intrinsic mobility as high as 5000 cm(2)/(V.s) can be obtained from such solution-processed graphene films when ionic screening is applied to nullify the Coulombic scattering by charged impurities. PMID- 20025235 TI - A multifrequency high-field electron paramagnetic resonance study of Co(II)S(4) coordination. AB - Advanced electron paramagnetic resonance (EPR) methods have been employed in the study of two high-spin cobalt(II) complexes, Co[(SPPh(2))(2)N](2) (Co(Ph,Ph)L(2)) and Co[(SPPh(2))(SP(i)Pr(2))N](2) (Co(iPr,Ph)L(2)), in which the bidentate disulfidoimidodiphosphinato ligands make up for a pseudotetrahedral sulfur coordination of the transition metal. The CoS(4) core in the two complexes has slightly different structure, owing to the different peripheral groups (phenyl or isopropyl) bound to the phosphorus atoms. To determine the zero-field splitting, notoriously difficult for high-spin cobalt(II), the two complexes required different approaches. For Co(Ph,Ph)L(2), the study of the X-band EPR spectrum of a single crystal as a function of temperature revealed a nearly axial character of the zero-field splitting (ZFS; E/D approximately -0.05). For Co(iPr,Ph)L(2), the combination of the EPR spectra at 9, 95, and 275 GHz revealed a rhombic character of the ZFS (E/D approximately -0.33). The energy difference between the Kramers doublets in Co(Ph,Ph)L(2) and Co(iPr,Ph)L(2) amounts to 24 cm(-1) and 30 cm(-1), respectively. From the X-band EPR spectra of diamagnetically diluted single crystals at fields up to 2.5 T for Co(Ph,Ph)L(2) and 0.5 T for Co(iPr,Ph)L(2), the effective g tensors and cobalt hyperfine tensors have been determined, including the direction of the principal axes in the cobalt sites. The values of the EPR observables are discussed in relation to the structural characteristics of the first (CoS(4)) and second coordination sphere in the complexes. PMID- 20025236 TI - Cytotoxic triterpene dilactones from the stems of Kadsura ananosma. AB - Six new triterpene dilactones with a rare rearranged pentacyclic skeleton, longipedlactones K-P (1-6), and seven known analogues (7-13) were isolated from the stems of Kadsura ananosma. Compound 1 was found to possess a unique peroxide bridge between C-1 and C-9 in rings A and B. The structures of these new compounds were established on the basis of spectroscopic data analysis, especially of their 2D NMR spectra. In the evaluation of the in vitro cytotoxicity of these compounds against a small panel of human cancer cell lines, compounds 3, 7, 9, and 13 were found to be the most potent against HL-60 acute leukemia cell. PMID- 20025237 TI - N-Heterocyclic carbene-catalyzed intramolecular aldehyde-nitrile cross coupling: an easy access to 3- aminochromones. AB - An immense effort has been made to develop an efficient strategy for the carbon carbon bond formation between aldehyde and nitrile intramolecularly using an N heterocyclic carbene catalyst to derive 3-aminochromone derivatives in good to excellent yields (80-95%). PMID- 20025238 TI - Singlet exciton fission for solar cell applications: energy aspects of interchromophore coupling. AB - Singlet exciton fission, a process that converts one singlet exciton to a pair of triplet excitons, has the potential to enhance the efficiency of both bulk heterojunction and dye-sensitized solar cells and is understood in crystals but not well understood in molecules. Previous studies have identified promising building blocks for singlet fission in molecular systems, but little work has investigated how these individual chromophores should be combined to maximize triplet yield. We consider the effects of chemically connecting two chromophores to create a coupled chromophore pair and compute how various structural choices alter the thermodynamic and kinetic parameters likely to control singlet fission yield. We use density functional theory to compute the electron transfer matrix element and the thermodynamics of fission for several promising chromophore pairs and find a trade-off between the desire to maximize this element and the desire to keep the singlet fission process exoergic. We identify promising molecular systems for singlet fission and suggest future experiments. PMID- 20025239 TI - States of adsorbed dodecyl amine and water at a silica surface as revealed by vibrational spectroscopy. AB - States of adsorbed dodecylamine (DDA) at a silica surface have been studied as a function of pH by vibrational spectroscopy (SFVS, FTIR), contact angle measurements, and molecular dynamics (MD) simulations. The results show that the state of adsorbed DDA at a silica surface varies significantly at different pH values. At pH 6.30, there is no pronounced adsorption of DDA cations and the surface is hydrophilic. At pH 10.0, the vibrational spectroscopy results together with contact angle measurements and MD simulations suggest that amine is adsorbed as a well organized monolayer, the hemimicelle structure. Under these conditions, dehydration occurs based on SFVS analysis and the silica surface becomes hydrophobic. In the case of pH 12.3, it has been confirmed that continued adsorption of DDA neutral molecules occurs with the amine surface state changing from a monolayer to a bilayer or a micellar surface state as revealed both from SFVS analysis and MD simulations. At this high pH, extensive surface hydration is evident from SFVS results and the silica surface becomes hydrophilic. PMID- 20025240 TI - Recognition by the thyroid hormone receptor of canonical DNA response elements. AB - To shed more light on the molecular requirements for recognition of thyroid response elements (TREs) by thyroid receptors (TRs), we compared the specific aspects of DNA TRE recognition by different TR constructs. Using fluorescence anisotropy, we performed a detailed and hierarchical study of TR-TRE binding. This was done by comparing the binding affinities of three different TR constructs for four different TRE DNA elements, including palindromic sequences and direct repeats (F2, PAL, DR-1, and DR-4) as well as their interactions with nonspecific DNA sequences. The effect of MgCl(2) on suppressing of nonselective DNA binding to TR was also investigated. Furthermore, we determined the dissociation constants of the hTRbeta DBD (DNA binding domain) and hTRbeta DBD LBD (DNA binding and ligand binding domains) for specific TREs. We found that a minimum DNA recognition peptide derived from DBD (H1TR) is sufficient for recognition and interaction with TREs, whereas scrambled DNA sequences were unrecognized. Additionally, we determined that the TR DBD binds to F2, PAL, and DR-4 with high affinity and similar K(d) values. The TR DBD-LBD recognizes all the tested TREs but binds preferentially to F2, with even higher affinity. Finally, our results demonstrate the important role played by LBDs in modulating TR-DNA binding. PMID- 20025241 TI - Neutron structure of human carbonic anhydrase II: implications for proton transfer. AB - Human carbonic anhydrase II (HCA II) catalyzes the reversible hydration of carbon dioxide to form bicarbonate and a proton. Despite many high-resolution X-ray crystal structures, mutagenesis, and kinetic data, the structural details of the active site, especially the proton transfer pathway, are unclear. A large HCA II crystal was prepared at pH 9.0 and subjected to vapor H-D exchange to replace labile hydrogens with deuteriums. Neutron diffraction studies were conducted at the Protein Crystallography Station at Los Alamos National Laboratory. The structure to 2.0 A resolution reveals several interesting active site features: (1) the Zn-bound solvent appearing to be predominantly a D(2)O molecule, (2) the orientation and hydrogen bonding pattern of solvent molecules in the active site cavity, (3) the side chain of His64 being unprotonated (neutral) and predominantly in an inward conformation pointing toward the zinc, and (4) the phenolic side chain of Tyr7 appearing to be unprotonated. The implications of these details are discussed, and a proposed mechanism for proton transfer is presented. PMID- 20025242 TI - Adventitious arsenate reductase activity of the catalytic domain of the human Cdc25B and Cdc25C phosphatases. AB - A number of eukaryotic enzymes that function as arsenate reductases are homologues of the catalytic domain of the human Cdc25 phosphatase. For example, the Leishmania major enzyme LmACR2 is both a phosphatase and an arsenate reductase, and its structure bears similarity to the structure of the catalytic domain of human Cdc25 phosphatase. These reductases contain an active site C-X(5) R signature motif, where C is the catalytic cysteine, the five X residues form a phosphate binding loop, and R is a highly conserved arginine, which is also present in human Cdc25 phosphatases. We therefore investigated the possibility that the three human Cdc25 isoforms might have adventitious arsenate reductase activity. The sequences for the catalytic domains of Cdc25A, -B, and -C were cloned individually into a prokaryotic expression vector, and their gene products were purified from a bacterial host using nickel affinity chromatography. While each of the three Cdc25 catalytic domains exhibited phosphatase activity, arsenate reductase activity was observed only with Cdc25B and -C. These two enzymes reduced inorganic arsenate but not methylated pentavalent arsenicals. Alteration of either the cysteine and arginine residues of the Cys-X(5)-Arg motif led to the loss of both reductase and phosphatase activities. Our observations suggest that Cdc25B and -C may adventitiously reduce arsenate to the more toxic arsenite and may also provide a framework for identifying other human protein tyrosine phosphatases containing the active site Cys-X(5)-Arg loop that might moonlight as arsenate reductases. PMID- 20025244 TI - Cross species quorum quenching using a native AI-2 processing enzyme. AB - Bacterial quorum sensing (QS) is a cell-cell communication process, mediated by signaling molecules, that alters various phenotypes including pathogenicity. Methods to interrupt these communication networks are being pursued as next generation antimicrobials. We present a technique for interrupting communication among bacteria that exploits their native and highly specific machinery for processing the signaling molecules themselves. Specifically, our approach is to bring native intracellular signal processing mechanisms to the extracellular surroundings and "quench" crosstalk among a variety of strains. In this study, the QS system based on the interspecies signaling molecule autoinducer-2 (AI-2) is targeted because of its prevalence among prokaryotes (it functions in over 80 bacterial species). We demonstrate that the Escherichia coli AI-2 kinase, LsrK, can phosphorylate AI-2 in vitro, and when LsrK-treated AI-2 is added ex vivo to E. coli populations, the native QS response is significantly reduced. Further, LsrK-mediated degradation of AI-2 attenuates the QS response among Salmonella typhimurium and Vibrio harveyi even though the AI-2 signal transduction mechanisms and the phenotypic responses are species-specific. Analogous results are obtained from a synthetic ecosystem where three species of bacteria (enteric and marine) are co-cultured. Finally, the addition of LsrK and ATP to growing co cultures of E. coli and S. typhimurium exhibits significantly reduced native "cross-talk" that ordinarily exists among and between species in an ecosystem. We believe this nature-inspired enzymatic approach for quenching QS systems will spawn new methods for controlling cell phenotype and potentially open new avenues for controlling bacterial pathogenicity. PMID- 20025243 TI - Hydrophobic residues in helix 8 of cannabinoid receptor 1 are critical for structural and functional properties. AB - In addition to the heptahelical transmembrane domain shared by all G protein coupled receptors (GPCRs), many class A GPCRs adopt a helical domain, termed helix 8, in the membrane-proximal region of the C terminus. We investigated the role of residues in the hydrophobic and hydrophilic faces of amphiphilic helix 8 of human cannabinoid receptor 1 (CB1). To differentiate between a role for specific residues and global features, we made two key mutants: one involving replacement of the highly hydrophobic groups, Leu404, Phe408, and Phe412, all with alanine and the second involving substitution of the basic residues, Lys402, Arg405, and Arg409, all with the neutral glutamine. The former showed a very low B(max) based on binding isotherms, a minimal E(max) based on GTPgammaS binding analysis, and defective localization relative to the wild-type CB1 receptor as revealed by confocal microscopy. However, the latter mutant and the wild-type receptors were indistinguishable. Circular dichroism spectroscopy of purified peptides with corresponding sequences indicated that the highly hydrophobic residues are critical for maintaining a strong helical structure in detergent, whereas the positively charged residues are not. Further investigation of mutant receptors revealed that CB1 localization requires a threshold level of hydrophobicity but not specific amino acids. Moreover, mutant receptors carrying two- to six-residue insertions amino-terminal to helix 8 revealed a graded decrease in B(max) values. Our results identify the key helix 8 components (including hydrophobicity of specific residues, structure, and location relative to TM7) determinant for receptor localization leading to robust ligand binding and G protein activation. PMID- 20025245 TI - Refining the in vitro and in vivo critical parameters for P-glycoprotein, [I]/IC50 and [I2]/IC50, that allow for the exclusion of drug candidates from clinical digoxin interaction studies. AB - The objective of this work was to further investigate the reasons for disconcordant clinical digoxin drug interactions (DDIs) particularly for false negative where in vitro data suggests no P-glycoprotein (P-gp) related DDI but a clinically relevant DDI is evident. Applying statistical analyses of binary classification and receiver operating characteristic (ROC), revised cutoff values for ratio of [I]/IC(50) < 0.1 and [I(2)]/IC(50) < 5 were identified to minimize the error rate, a reduction of false negative rate to 9% from 36% (based on individual ratios). The steady state total C(max) at highest dose of the inhibitor is defined as [I] and the ratio of the nominal maximal gastrointestinal concentration determined for highest dose per 250 mL volume defined [I(2)](.) We also investigated the reliability of the clinical data to see if recommendations can be made on values that would allow predictions of 25% change in digoxin exposure. The literature derived clinical digoxin interaction studies were statistically powered to detect relevant changes in exposure associated with digitalis toxicities. Our analysis identified that many co-meds administered with digoxin are cardiovascular (CV) agents. Moreover, our investigations also suggest that the presence of CV agents may alter cardiac output and/or kidney function that may act alone or are additional components to enhance digoxin exposure along with P-gp interaction. While we recommend digoxin as the probe substrate to define P-gp inhibitory potency for clinical assessment, we observed high concordance in P-gp inhibitory potency for calcein AM as a probe substrate. PMID- 20025246 TI - Measurement of intrinsic rate constants in the tyrosine hydroxylase reaction. AB - Tyrosine hydroxylase (TyrH) is a pterin-dependent mononuclear non-heme aromatic amino acid hydroxylase that catalyzes the conversion of tyrosine to dihydroxyphenylalanine (DOPA). Chemical quench analyses of the enzymatic reaction show a burst of DOPA formation, followed by a linear rate equal to the k(cat) value at both 5 and 30 degrees C. The effects of increasing solvent viscosity confirm that k(cat) is approximately 84% limited by diffusion, most probably due to slow product release, and that tyrosine has a commitment to catalysis of 0.45. The effect of viscosity on the k(cat)/K(m) for 6-methyltetrahydropterin is greater than the theoretical limit, consistent with the coupling of pterin binding to the movement of a surface loop. The absorbance changes in the spectrum of the tetrahydropterin during the first turnover, the kinetics of DOPA formation during the first turnover, and the previously described kinetics for formation and decay of the Fe(IV)O intermediate [Eser, B. E., Barr, E. W., Frantom, P. A., Saleh, L., Bollinger, J. M., Jr., Krebs, C., and Fitzpatrick, P. F. (2007) J. Am. Chem. Soc. 129, 11334-11335] were analyzed globally, yielding a single set of rate constants for the TyrH reaction. Reversible binding of oxygen is followed by formation of Fe(IV)O and 4a-hydroxypterin with a rate constant of 13 s(-1) at 5 degrees C. Transfer of oxygen from Fe(IV)O to tyrosine to form DOPA follows with a rate constant of 22 s(-1). Release of DOPA and/or the 4a-hydroxypterin with a rate constant of 0.86 s(-1) completes the turnover. PMID- 20025247 TI - Mapping of the signal peptide-binding domain of Escherichia coli SecA using Forster resonance energy transfer. AB - Identification of the signal peptide-binding domain within SecA ATPase is an important goal for understanding the molecular basis of SecA preprotein recognition as well as elucidating the chemo-mechanical cycle of this nanomotor during protein translocation. In this study, Forster resonance energy transfer methodology was employed to map the location of the SecA signal peptide-binding domain using a collection of functional monocysteine SecA mutants and alkaline phosphatase signal peptides labeled with appropriate donor-acceptor fluorophores. Fluorescence anisotropy measurements yielded an equilibrium binding constant of 1.4 or 10.7 muM for the alkaline phosphatase signal peptide labeled at residue 22 or 2, respectively, with SecA, and a binding stoichiometry of one signal peptide bound per SecA monomer. Binding affinity measurements performed with a monomer biased mutant indicate that the signal peptide binds equally well to SecA monomer or dimer. Distance measurements determined for 13 SecA mutants show that the SecA signal peptide-binding domain encompasses a portion of the preprotein cross linking domain but also includes regions of nucleotide-binding domain 1 and particularly the helical scaffold domain. The identified region lies at a multidomain interface within the heart of SecA, surrounded by and potentially responsive to domains important for binding nucleotide, mature portions of the preprotein, and the SecYEG channel. Our FRET-mapped binding domain, in contrast to the domain identified by NMR spectroscopy, includes the two-helix finger that has been shown to interact with the preprotein during translocation and lies at the entrance to the protein-conducting channel in the recently determined SecA SecYEG structure. PMID- 20025249 TI - Jasmonates: structural requirements for lipid-derived signals active in plant stress responses and development. AB - Jasmonates are lipid-derived signals that mediate plant stress responses and development processes. Enzymes participating in biosynthesis of jasmonic acid (JA) (1, 2) and components of JA signaling have been extensively characterized by biochemical and molecular-genetic tools. Mutants of Arabidopsis and tomato have helped to define the pathway for synthesis of jasmonoyl-isoleucine (JA-Ile), the active form of JA, and to identify the F-box protein COI1 as central regulatory unit. However, details of the molecular mechanism of JA signaling have only recently been unraveled by the discovery of JAZ proteins that function in transcriptional repression. The emerging picture of JA perception and signaling cascade implies the SCF(COI1) complex operating as E3 ubiquitin ligase that upon binding of JA-Ile targets JAZ repressors for degradation by the 26S-proteasome pathway, thereby allowing the transcription factor MYC2 to activate gene expression. The fact that only one particular stereoisomer, (+)-7-iso-JA-l-Ile (4), shows high biological activity suggests that epimerization between active and inactive diastereomers could be a mechanism for turning JA signaling on or off. The recent demonstration that COI1 directly binds (+)-7-iso-JA-l-Ile (4) and thus functions as JA receptor revealed that formation of the ternary complex COI1 JA-Ile-JAZ is an ordered process. The pronounced differences in biological activity of JA stereoisomers also imply strict stereospecific control of product formation along the JA biosynthetic pathway. The pathway of JA biosynthesis has been unraveled, and most of the participating enzymes are well-characterized. For key enzymes of JA biosynthesis the crystal structures have been established, allowing insight into the mechanisms of catalysis and modes of substrate binding that lead to formation of stereospecific products. PMID- 20025248 TI - Cells on pores: a simulation-driven analysis of transcellular small molecule transport. AB - A biophysical, computational model of cell pharmacokinetics (1CellPK) is being developed to enable prediction of the intracellular accumulation and transcellular transport properties of small molecules using their calculated physicochemical properties as input. To test if 1CellPK can generate accurate, quantitative hypotheses and guide experimental analysis of the transcellular transport kinetics of small molecules, epithelial cells were grown on impermeable polyester membranes with cylindrical pores and chloroquine (CQ) was used as a transport probe. The effect of the number of pores and their diameter on transcellular transport of CQ was measured in apical-to-basolateral or basolateral-to-apical directions, at pH 7.4 and 6.5 in the donor compartment. Experimental and simulation results were consistent with a phospholipid bilayer limited, passive diffusion transport mechanism. In experiments and 1CellPK simulations, intracellular CQ mass and the net rate of mass transport varied <2 fold although total pore area per cell varied >10-fold, so by normalizing the net rate of mass transport by the pore area available for transport, cell permeability on 3 mum pore diameter membranes was more than an order of magnitude less than on 0.4 mum pore diameter membranes. The results of simulations of transcellular transport were accurate for the first four hours of drug exposure, but those of CQ mass accumulation were accurate only for the first five minutes. Upon prolonged incubation, changes in cellular parameters such as lysosome pH rise, lysosome volume expansion, and nuclear shrinkage were associated with excess CQ accumulation. Based on the simulations, lysosome volume expansion alone can partly account for the measured, total intracellular CQ mass increase, while adding the intracellular binding of the protonated, ionized forms of CQ (as reflected in the measured partition coefficient of CQ in detergent-permeabilized cells at physiological pH) can further improve the intracellular CQ mass accumulation prediction. PMID- 20025250 TI - Nonlinear optical imaging of integral membrane protein crystals in lipidic mesophases. AB - Second-order nonlinear optical imaging of chiral crystals (SONICC) is explored for selective detection of integral membrane protein crystals grown in opaque and turbid environments. High turbidity is a hallmark of membrane protein crystallization due to the extensive use of detergent and/or lipids that often form various mesophases. Detection of crystals in such media by conventional optical methods (e.g., intrinsic UV fluorescence, birefringence, bright-field image analysis, etc.) is often complicated by optical scattering and by the small sizes of the crystals that routinely form. SONICC is shown to be well-suited for this application, by nature of its compatibility with imaging in scattering media and its high selectivity for protein crystals. Bright second harmonic generation (SHG) (up to 18 million counts/s) was observed from even relatively small crystals (5 mum) with a minimal background due to the surrounding lipid mesophase ( approximately 1 thousand counts/s). The low background nature of the resulting protein crystal images permitted the use of a relatively simple, particle counting analysis for preliminary scoring. Comparisons between a particle counting analysis of SONICC images and protocols based on the human expert analysis of conventional bright-field and birefringence images were performed. PMID- 20025251 TI - Regioselective halogenations and subsequent Suzuki-Miyaura coupling onto bicyclic 2-pyridones. AB - A selective synthesis of 6-bromo-8-iodo dihydro thiazolo ring-fused 2-pyridones is described. These halogenated 2-pyridones are selectively arylated by sequential Suzuki-Miyaura couplings. This approach can advantageously be used to synthesize focused libraries of substituted ring-fused 2-pyridones, a class of compounds with novel antibacterial properties. PMID- 20025252 TI - Temperature-sensitive phase transition of dendritic polyethylene amphiphiles with core-shell architecture revealed by a Rayleigh scattering technique. AB - The phase-transition behavior of unimolecular dendritic polyethylene amphiphiles with core-shell architecture aqueous solutions was investigated by a Rayleigh scattering (RS) technique. Dendritic polyethylene (DPE) poly(oligo(ethylenegylcol) methacrylate) (POEGMA) with a DPE hydrophobic core and a POEGMA hydrophilic shell was synthesized by the atom-transfer radical polymerization (ATRP) of OEGMA using DPE terminated by the bromine group as a macroinitiator. The fluorescence measurements implied that DPE-POEGMA molecules in aqueous solutions existed as the unimolecular micelles. To understand the phase-transition behavior of dendritic polyethylene amphiphilic unimolecular micelles in aqueous solutions, the temperature dependence of the RS spectra of DPE-POEGMA aqueous solutions under the heating-and-cooling cycle indicated that the heating and cooling processes were reversible but hysteresis existed. The phase transition of DPE-POEGMA aqueous solutions decelerated with increasing levels of PEGylation. DPE-POEGMA exhibited a lower phase-transition temperature in D(2)O than in water. PMID- 20025253 TI - Stable silver(I) hydride complexes supported by diselenophosphate ligands. AB - The first stable structure of silver(I) cluster cations [Ag(8)(mu(4) H){Se(2)P(OR)(2)}(6)](+) [R = (i)Pr, 1; Et, 2] containing Ag(I)-hydride bridges (Ag-mu-H-Ag) in T symmetry was reported. The clusters having an interstitial hydride were composed of an octanuclear silver core in tetracapped tetrahedral geometry, which was inscribed within a Se(12) icosahedron represented by six dialkyl diselenophosphate ligands in a tetrametallic-tetraconnective (mu(2), mu(2)) bonding mode. The presence of hydride was unequivocally corroborated by both (1)H and (109)Ag NMR spectroscopies of which a nonet in the (1)H NMR spectrum for the hydride resonance coupled with a doublet peak observed in the (109)Ag NMR spectrum clearly suggests that eight silver nuclei are equivalent in the NMR time scale and a fast exchange of the positions between the vertex and capping silver atoms in solution must occur. The hypothesis was also supported by a density functional theory (DFT) investigation on a simplified model [Ag(8)(H)(Se(2)PH(2))(6)](+), which confirmed that the Ag(8)H cubic core of T(h) symmetry may not be formed as it is energetically highly unfavorable (0.67 eV less stable than the T structure). PMID- 20025254 TI - Copper(II) and nickel(II) complexes of beta-aminoketoxime ligand: syntheses, crystal structures, magnetism, and nickel(II) templated coupling of oxime with nitrile. AB - The syntheses, molecular structures, and magnetic properties of a dicopper(II) complex, [Cu(2)(HL(1))(2)](ClO(4))(2) (1), and its nickel(II) analog, [Ni(2)(HL(1))(2)](ClO(4))(2) (2), of a beta-amino ketoxime ligand (H(2)L(1) = 4,4,9,9-tetramethyl-5,8-diazadodecane-2,11-dione dioxime) are discussed. The metal centers in out-of-plane oximate bridged dinuclear complexes (1 and 2) display distorted trigonal bipyramidal geometry and form a six-membered M(2)(NO)(2) ring oriented in a boat conformation. The two copper(II) centers in 1 interact ferromagnetically giving rise to a triplet-spin ground state whereas the two nickel(II) centers in 2 interact antiferromagnetically to stabilize a singlet spin state. Variable temperature magnetic susceptibility measurements establish the presence of a weak ferromagnetic coupling (J = 13 cm(-1)) in 1 and a weak anitiferromagnetic coupling (J = -12 cm(-1)) in 2. The exchange coupling constant derived from B3LYP computations in conjunction with broken symmetry spin projection techniques for the oximate bridged dinuclear copper(II) complex shows excellent agreement with the corresponding experimental value. A square-planar mononuclear nickel(II) complex of the dioxime ligand, [Ni(H(2)L(1))](ClO(4))(2) (3), is reported along with its crystal structure, which reacts with acetonitrile to produce a six-coordinate mononuclear complex, [Ni(L(2))](ClO(4))(2) (4). The ligand (L(2)) in complex 4 is the iminoacyl derivative of oxime, where the coupling of oxime and acetonitrile takes place via a proton-assisted pathway. The iminoacylation of H(2)L(1) works with other nitriles like butyronitrile and benzonitrile. Computational studies support a proton-assisted coupling of oxime with nitrile. The critical transition states have been located for the iminoacylation reaction. Complex 4 can be converted back to complex 3 by reacting with sodium acetate in methanol. PMID- 20025255 TI - Pressure and temperature dependence of ethyl nitrate formation in the C(2)H(5)O(2) + NO reaction. AB - The branching ratio beta = k(1b)/k(1a) for the formation of ethyl nitrate, C(2)H(5)ONO(2), in the gas-phase C(2)H(5)O(2) + NO reaction, C(2)H(5)O(2) + NO - > C(2)H(5)O + NO(2) (1a), C(2)H(5)O(2) + NO --> C(2)H(5)ONO(2) (1b), was determined over the pressure and temperature ranges 100-600 Torr and 223-298 K, respectively, using a turbulent flow reactor coupled with a chemical ionization mass spectrometer. At 298 K the C(2)H(5)ONO(2) yield was found to increase linearly with pressure from about 0.7% at 100 Torr to about 3% at 600 Torr. At each pressure, the branching ratio of C(2)H(5)ONO(2) formation increases with the decrease of temperature. The following parametrization equation has been derived in the pressure and temperature ranges of the study: beta(P,T) (%) = (3.88 x 10( 3).P (Torr) + 0.365).(1 + 1500(1/T - 1/298)). The atmospheric implication of the results obtained is briefly discussed, in particular the impact of beta on the evolution of ethyl nitrate in urban plumes. PMID- 20025256 TI - Ligand-controlled enantioselective [2 + 2] cycloaddition of oxabicyclic alkenes with terminal alkynes using chiral iridium catalysts. AB - The first catalytic asymmetric [2 + 2] cycloaddition of oxabicyclic alkenes and terminal alkynes has been developed. This iridium-catalyzed enantioselective [2 + 2] cycloaddition allows the formation of four stereocenters in a single step with excellent enantioselectivity (94-->99% ee). PMID- 20025257 TI - Synthesis of pyrroles by gold(I)-catalyzed amino-claisen rearrangement of N propargyl enaminone derivatives. AB - The cationic N-heterocyclic carbene-gold(I) complex catalyzes the formation of tri- and tetrasubstituted pyrroles via the amino-Claisen rearrangement of N propargyl beta-enaminone derivatives and the cyclization of alpha-allenyl beta enaminone intermediates. PMID- 20025259 TI - Analyses of the infrared absorption bands of (15)NO(3) in the 1850-3150 cm(-1) region. AB - We have observed the infrared spectrum of (15)NO(3) by a high resolution Fourier transform infrared (FT-IR) spectrometer using the reaction of F atoms with H(15)NO(3). Five (2)E'-(2)A(2)' bands are identified in the 1850-3150 cm(-1) region. The rotational analyses indicate that these bands have the lower state in common, which coincides with the ground state of planar D(3h) symmetry. The upper (2)E' states more or less suffer from perturbations by close-lying dark states. Among them, those of the 2004, 2128, and 2492 cm(-1) bands are analyzed to determine molecular parameters in these states by fixing the ground-state constants to those derived by a combination difference method. The spin-orbit and Coriolis coupling constants in the (2)E' states are substantially different for different vibronic states. The vibrational assignments of NO(3) in the ground electronic state are discussed using experimental data heretofore available, supplemented by those obtained by the present study. PMID- 20025258 TI - Toward designed singlet fission: electronic states and photophysics of 1,3 diphenylisobenzofuran. AB - Single crystal molecular structure and solution photophysical properties are reported for 1,3-diphenylisobenzofuran (1), of interest as a model compound in studies of singlet fission. For the ground state of 1 and of its radical cation (1(+*)) and anion (1(-*)), we report the UV-visible absorption spectra, and for neutral 1, also the magnetic circular dichroism (MCD) and the decomposition of the absorption spectrum into purely polarized components, deduced from fluorescence polarization. These results were used to identify a series of singlet excited states. For the first excited singlet and triplet states of 1, the transient visible absorption spectra, S(1) --> S(x) and sensitized T(1) --> T(x), and single exponential lifetimes, tau(F) = approximately 5.3 ns and tau(T) = approximately 200 micros, are reported. The spectra and lifetimes of S(1) --> S(0) fluorescence and sensitized T(1) --> T(x) absorption of 1 were obtained in a series of solvents, as was the fluorescence quantum yield, Phi(F) = 0.95-0.99. No phosphorescence has been detected. The first triplet excitation energy of solid 1 (11,400 cm(-1)) was obtained by electron energy loss spectroscopy, in agreement with previously reported solution values. The fluorescence excitation spectrum suggests an onset of a nonradiative channel at approximately 37,000 cm(-1). Excitation energies and relative transition intensities are in agreement with those of ab initio (CC2) calculations after an empirical 3000 cm(-1) adjustment of the initial state energy to correct differentially for a better quality description of the initial relative to the terminal state of an absorption transition. The interpretation of the MCD spectrum used the semiempirical PPP method, whose results for the S(0) --> S(x) spectrum require no empirical adjustment and are otherwise nearly identical with the CC2 results in all respects including the detailed nature of the electronic excitation. The ground state geometry of 1 was also calculated by the MP2, B3LYP, and CAS methods. The calculations provided a prediction of changes of molecular geometry upon excitation or ionization and permitted an interpretation of the spectra in terms of molecular orbitals involved. Computations suggest that 1 can exist as two nearly isoenergetic conformers of C(2) or C(s) symmetry. Linear dichroism measurements in stretched polyethylene provide evidence for their existence and show that they orient to different degrees, permitting a separation of their spectra in the region of the purely polarized first absorption band. Their excitation energies are nearly identical, but the Franck-Condon envelopes of their first transition differ to a surprising degree. PMID- 20025260 TI - Strategies and limitations in dendrimeric immunogen synthesis. The influenza virus M2e epitope as a case study. AB - Dendrimeric platforms such as multiple antigen peptides (MAPs) are regarded as one of the most efficacious approaches for antigenic presentation. Originally described as available by stepwise solid-phase peptide synthesis (SPPS), MAPs have also been prepared by chemical (thioether, oxime, hydrazone) ligation of appropriately functionalized tetra- or octavalent polylysine scaffolds with the peptide antigen to be multiply displayed. In this work, the advantages and limitations of two of the most frequent methods of MAP preparation, namely, chemoselective thioether ligation in solution, and all-solid-phase synthesis, have been tested in the case of a particularly troublesome epitope model, the ectodomain of protein M2 from influenza virus (M2e). The strong tendency of M2e to self-associate is a serious inconvenient for conjugation in solution, which as a result fails to produce the target MAPs with the specified number of M2e copies. In contrast, the fully stepwise SPPS approach is shown to be quite practical, especially when 6-aminohexanoic acid spacer units providing increased internal flexibility are inserted at each branching point. PMID- 20025261 TI - Impact of drying on wood ultrastructure observed by deuterium exchange and photoacoustic FT-IR spectroscopy. AB - The impact of drying on the ultrastructure of fresh wood was studied by deuterium exchange coupled with FT-IR analysis. This fundamental investigation demonstrated that water removal leads to irreversible alterations of the wood structure, namely, supramolecular rearrangements between wood polymers. The deuteration of fresh wood was shown to be fully reversible by a subsequent exposure of the deuterated sample to water (reprotonation). Therefore, the presence of any OD groups in deuterated and then dried wood samples after reprotonation is a clear indicator of reduced accessibility. The extent of changes was affected by drying temperature and relative humidity. Application of this methodology for the evaluation of chemical pulp sample (reference material) resulted in similar response, only more pronounced. Two hypothetical alternatives were proposed for accessibility reduction in dried wood: (i) irreversible aggregation of cellulose microfibrils and (ii) irreversible stiffening of the hemicellulose/lignin matrix that extensively swells when exposed to water. PMID- 20025262 TI - Kinetic evidence for dihapto (eta(2)) pi-aryl participation in acid-catalyzed ring opening of diarylhomobenzoquinone epoxides. AB - The BF(3)-catalyzed ring-opening reaction of variously endo/exo m- and p substituted diarylhomobenzoquinone epoxides proceeded through a transannular S(E)2-Ar cyclization of endo-aryl groups to give the tricyclic diketo-alcohols and cyclohexadienone spiro-linked tricyclic diketo-alcohols. Kinetics of these reactions has been investigated in CDCl(3) at 30 degrees C in order to elucidate the possible remote pi-aryl participation. The rates were significantly increased with increasing electron-donating ability of the endo-aryl substituents X (k(p MeO)/k(p-CF3) = 8200) but only negligibly influenced by the distal exo-aryl substituents Y (k(p-MeO)/k(p-CF3) = 2.1). For the endo-X substituted series, an excellent linear free energy relationship, log k(rel)(endo) = -2.49sigma(ipso) - 1.62sigma(ortho) - 0.108 (R(2) = 0.98, n = 8), was attained using two modified site-dependent substituent parameters sigma(ipso) (using sigma(p)(+) for p-X and sigma(m) for m-X) and sigma(ortho) (using sigma(m) for p-X and sigma(p)(+) for m X). This means that the dihapto(eta(2)) pi-coordination occurs in the pi-aryl participation, with the ipso pi-electron donation contributing 1.6 times more effectively than the ortho one. On the other hand, the distal exo-Y substituted series gave an acceptable Yukawa-Tsuno equation with small polar and resonance contributions; log k(rel)(exo) = -0.912(sigma(0) + 0.237Deltasigma(R)(+)) (R(2) = 0.96, n = 8). These kinetic substituent effects were compared with those of the acid-catalyzed pi-aryl assisted transannular S(E)2-Ar cyclization of the cyclobutene-fused diarylhomobenzoquinones. It was found that the geometrical characteristics of the vacant oxirane Walsh orbital and the cyclobutene antibonding orbital play a crucial role in the topological features of eta(2) pi aryl participation. PMID- 20025263 TI - pH-Dependent syntheses and crystal structures of a series of organic-inorganic hybrids constructed from Keggin or Wells-Dawson polyoxometalates and silver coordination compounds. AB - A series of silver complexes of polyoxometalates (POMs), formulated as, [Ag(2)(4,4'-bpy)(2)(4,4'-Hbpy)(H(2)O)][PW(12)O(40)] (4,4'-bpy = 4,4'-bipyridine, 1), [Ag(4)(4,4'-bpy)(4)][4,4'-H(2)bpy][P(2)W(18)O(62)].4H(2)O (2), [Ag(4)(4,4' bpy)(4)][H(2)P(2)W(18)O(62)].7H(2)O (3), and Na(8)[Ag(3)(4,4' bpy)(3)][PW(10)Ag(2)O(39)].6H(2)O (4), have been hydrothermally synthesized in situ at different pH. Complex 1, based on saturated Keggin POM building blocks and silver coordination compounds, exhibits an interesting 3-fold interpenetration of diamondlike network in the POM chemistry. Different from complex 1, complexes 2-3 consist of Wells-Dawson polyoxoanions and silver coordination compounds. Complex 2 represents the highest coordination number of Wells-Dawson polyoxoanions, which helps to form a high-dimensional framework with (4.6(4).8)(4)(4(4).6(16).8(8)) topology. Thee-dimensional (3D) Wells-Dawson phosphotungstate 3 reveals another new topology (4.6(4).7)(4.6(4).8)(2)(4(3).6(8).7(2).8(2)). Complex 4 forms a 3D framework constructed from divacant Keggin polyoxoanions and silver coordination compounds. Their structural differences indicate that the pH value of the reaction system plays a key role on the structures and topologies of these complexes, and the whole self-assembly process is pH-dependent. PMID- 20025264 TI - An efficient stereoselective synthesis of penaresidin a from (e)-2-protected amino-3,4-unsaturated sulfoxide. AB - An efficient, modular, asymmetric synthesis of penaresidin A is disclosed. A beta protected amino-gamma,delta-unsaturated sulfoxide was prepared by stereoselective addition of the lithio anion of (R)-methyl p-tolyl sulfoxide to an unsaturated sulfinylimine. The pendant sulfoxide group was used as an intramolecular nucleophile to functionalize an alkene regio- and stereoselectively to furnish a bromohydrin, which was employed as the key intermediate in the preparation of the azetidine subunit of penaresidin A. The stereogenic centers of the side chain were introduced by a regioselective opening of an epoxide. Julia-Kocienski olefination was used to couple the azetidine and side chain subunits. The methodology disclosed herein is also useful for the synthesis of ribo- and arabino-phytosphingosines and compounds possessing the amino alcohol moiety. PMID- 20025265 TI - Molecular dynamics studies of concentrated binary aqueous solutions of lanthanide salts: structures and exchange dynamics. AB - Concentrated binary aqueous solutions of lanthanide (Nd(3+) and Dy(3+)) salts (ClO(4)(-), Cl(-), and NO(3)(-)) have been studied by means of classical molecular dynamics (MD) simulations with explicit polarization and UV-visible spectroscopy. Pair interaction potentials, used for the MD simulations, have been developed in order to reproduce experimental hydration properties. Nd(3+) and Dy(3+) have been chosen because of their position in the lanthanide series: Nd(3+) being a light lanthanide and Dy(3+) a heavy one. They are respectively coordinated to nine and eight water molecules, in pure water, involving changes in their salt hydration structures. Both MD simulations and UV-visible experiments highlight the stronger affinity of nitrate anions toward Ln(3+) compared to perchlorates and chlorides. Dissociation/association processes of Nd(3+)-Cl(-) and Nd(3+)-NO(3)(-) ion pairs in aqueous solution have been analyzed using potential of mean force profile calculations. Furthermore, from MD simulations, it appears that the affinity of anions (perchlorate, chloride, and nitrate) is stronger for Nd(3+) than Dy(3+). PMID- 20025266 TI - Influence of conformation on conductance of biphenyl-dithiol single-molecule contacts. AB - The conductance of a family of biphenyl-dithiol derivatives with conformationally fixed torsion angle was measured using the scanning tunneling microscopy (STM) break-junction method. We found that it depends on the torsion angle phi between two phenyl rings; twisting the biphenyl system from flat (phi = 0 degrees ) to perpendicular (phi = 90 degrees ) decreased the conductance by a factor of 30. Detailed calculations of transport based on density functional theory and a two level model (TLM) support the experimentally obtained cos(2) phi correlation between the junction conductance G and the torsion angle phi. The TLM describes the pair of hybridizing highest occupied molecular orbital (HOMO) states on the phenyl rings and illustrates that the pi-pi coupling dominates the transport under "off-resonance" conditions where the HOMO levels are well separated from the Femi energy. PMID- 20025267 TI - Sandwiched graphene--membrane superstructures. AB - We demonstrate by molecular dynamics simulations that graphene sheets could be hosted in the hydrophobic interior of biological membranes formed by amphiphilic phospholipid molecules. Our simulation shows that these hybrid graphene--membrane superstructures might be prepared by forming hydrated micelles of individual graphene flakes covered by phospholipids, which can be then fused with the membrane. Since the phospholipid layers of the membrane electrically isolate the embedded graphene from the external solution, the composite system might be used in the development of biosensors and bioelectronic materials. PMID- 20025268 TI - Highly enantioselective titanium-catalyzed cyanation of imines at room temperature. AB - A highly active and enantioselective titanium-catalyzed cyanation of imines at room temperature is described. The catalyst used is a partially hydrolyzed titanium alkoxide (PHTA) precatalyst together with a readily available N-salicyl beta-aminoalcohol ligand. Up to 98% ee was obtained with quantitative yields in 15 min of reaction time using 5 mol % of the catalyst. Various N-protecting groups such as benzyl, benzhydryl, Boc, and PMP are tolerated. PMID- 20025269 TI - Antibacterial effects of poly(2-(dimethylamino ethyl)methacrylate) against selected gram-positive and gram-negative bacteria. AB - Antimicrobial coatings can reduce the occurrence of medical device-related bacterial infections. Poly(2-(dimethylamino ethyl)methacrylate) (pDMAEMA) is one such polymer that is being researched in this regard. The aims of this study were to (1) elucidate pDMAEMA's antimicrobial activity against a range of Gram positive and Gram-negative bacteria and (2) to investigate its antimicrobial mode of action. The methods used include determination of minimum inhibitory concentration (MIC) values against various bacteria and the effect of pH and temperature on antimicrobial activity. The ability of pDMAEMA to permeabilise bacterial membranes was determined using the dyes 1-N-phenyl-naphthylamine and calcein-AM. Flow cytometry was used to investigate pDMAEMA's capacity to be internalized by bacteria and to determine effects on bacterial cell cycling. pDMAEMA was bacteriostatic against Gram-negative bacteria with MIC values between 0.1-1 mg/mL. MIC values against Gram-positive bacteria were variable. pDMAEMA was active against Gram-positive bacteria around its pK(a) and at lower pH values, while it was active against Gram-negative bacteria around its pK(a) and at higher pH values. pDMAEMA inhibited bacterial growth by binding to the outside of the bacteria, permeabilizing the outer membrane and disrupting the cytoplasmic membrane. By incorporating pDMAEMA with erythromycin, it was found that the efficacy of the latter was increased against Gram-negative bacteria. Together, the results illustrate that pDMAEMA acts in a similar fashion to other cationic biocides. PMID- 20025270 TI - Synthesis and characterization of bionanocomposites with tunable properties from poly(lactic acid) and acetylated microfibrillated cellulose. AB - In the present study, novel bionanocomposite materials with tunable properties were successfully prepared using a poly(lactic acid) (PLA) matrix and acetylated microfibrillated cellulose (MFC) as reinforcing agent. The acetylation of MFC was confirmed by FTIR and (13)C CP-MAS NMR spectroscopies. The grafting of acetyl moieties on the cellulose surface not only prevented MFC hornification upon drying but also dramatically improved redispersibility of the powdered nanofibers in chloroform, a PLA solvent of low polarity. Moreover, we demonstrate that the properties of the resulting PLA nanocomposites could be tailored by adjusting both the acetyl content (Ac%) and the amount of MFC. These nanomaterials showed improved filler dispersion, higher thermal stability, and reduced hygroscopicity with respect to those prepared with unmodified MFC. Dynamic mechanical analysis (DMA) highlighted the reinforcing potential of both the unmodified and the acetylated MFC on the viscoelastic properties of the neat PLA. But more interesting, an increase in the PLA glass transition temperature was detected when using the 8.5% acetylated MFC at 17 wt %, indicating an improved compatibility at the fiber-matrix interface. These findings suggest that the final properties of nanocomposite materials can be controlled by adjusting the %Ac of MFC. PMID- 20025271 TI - Synthesis and characterization of stigmasterol oxidation products. AB - The synthesis and structural characterization of a series of oxides of stigmasterol is described providing a valuable series of reference standards for these oxides, analogous to the cholesterol oxidation products (COPs) which have been shown to have detrimental biological effects. Biological evaluation of the oxides of phytosterols is significant in the context of increased dietary use of phytosterols in the drive to reduce cholesterol absorption. PMID- 20025272 TI - Delphinidin induces necrosis in hepatocellular carcinoma cells in the presence of 3-methyladenine, an autophagy inhibitor. AB - The present study was performed to determine whether anthocyanins could trigger different modes of cell death in different cancers. It was found that whereas cyanidin-3-rutinoside and delphinidin could induce apoptosis in leukemia cells, they caused growth retardation in hepatocellular carcinoma cells (HCC), which was accompanied with a significant cellular vacuolization. The latter was likely caused by macroautophagy and was completely suppressed by 3-methyladenine, an inhibitor of class III phosphoinositide 3-kinase that is important for autophagy activation, and by bafilomycin A1, which blocks lysosomal degradation. Delphinidin induced significant lipidation of LC3, an indication of macroautophagy, which was also suppressed by 3-methyladenine. Macroautophagy was required for the survival of delphinidin-treated HCC cells as inhibition with 3 methyladenine led to massive necrosis without caspase activation. Thus, anthocyanins could induce different modes of cell death for different cancers. Furthermore, anthocyanins could be used in combination with a macroautophagy inhibitor for treating cancers such as HCC. PMID- 20025273 TI - Ecotoxicity by the biodegradation of alkylphenol polyethoxylates depends on the effect of trace elements. AB - The bacteria Sphingomonas sp. strain BSN22, isolated from bean fields, degraded octylphenol polyethoxylates (OPEO(n)) to octylphenol (OP) under aerobic conditions. This biodegradation mechanism proceeded by the following two-step degradation process: (1) degradation of OPEO(n) to octylphenol triethoxylate (OPEO(3)), (2) degradation from OPEO(3) to OP via octylphenoxy acetic acid (OPEC(1)). The chemical structure of OPEC(1) was confirmed by analysis using (18)O-labeled water. Quantitative studies revealed that magnesium (Mg(2+)) and calcium (Ca(2+)) ions were essential for the biodegradation of OPEO(n). Furthermore, the rate of biodegradation was especially accelerated by ferric ions (Fe(3+)), and the accumulated amounts of endocrine active chemicals, such as OP, OPEO(1), and OPEC(1), significantly increased to the concentration of 22.8, 221.7, and 961.1 microM in the presence of 37.0 microM Fe(3+), respectively. This suggests that environmental elements significantly influence the resultant ecotoxicity as well as the rate of their biodegradation in the environment. This study on the mechanism of OPEO(n) biodegradation may play an important role in understanding and managing environmental safety, including drinking water safety. PMID- 20025274 TI - Simultaneous stable carbon isotopic analysis of wine glycerol and ethanol by liquid chromatography coupled to isotope ratio mass spectrometry. AB - A novel procedure was established for the simultaneous characterization of wine glycerol and ethanol (13)C/(12)C isotope ratio, using liquid chromatography/isotope ratio mass spectrometry (LC-IRMS). Several parameters influencing separation of glycerol and ethanol from wine matrix were optimized. Results obtained for 35 Spanish samples exposed no significant differences and very strong correlations (r = 0.99) between the glycerol (13)C/(12)C ratios obtained by an alternative method (gas chromatography/isotope ratio mass spectrometry) and the proposed new methodology, and between the ethanol (13)C/(12)C ratios obtained by the official method (elemental analyzer/isotope ratio mass spectrometry) and the proposed new methodology. The accuracy of the proposed method varied from 0.01 to 0.19 per thousand, and the analytical precision was better than 0.25 per thousand. The new developed LC-IRMS method it is the first isotopic method that allows (13)C/(12)C determination of both analytes in the same run directly from a liquid sample with no previous glycerol or ethanol isolation, overcoming technical difficulties associated with complex sample treatment and improving in terms of simplicity and speed. PMID- 20025275 TI - HPLC assay for characterizing alpha-cyano-3-phenoxybenzyl pyrethroids hydrolytic metabolism by Helicoverpa armigera (Hubner) based on the quantitative analysis of 3-phenoxybenzoic acid. AB - A new HPLC assay for characterizing the hydrolytic metabolism of alpha-cyano-3 phenoxybenzyl pyrethroids in Helicoverpa armigera (Hubner) through quantitative analysis of the common metabolite 3-phenoxybenzoic acid (3-PBA), converted from 3 phenoxybenzaldehyde (3-PBAld) by aldehyde-oxidizing enzymes, has been established. The method was validated by measuring beta-cypermethrin metabolism in H. armigera. The hydrolytic activity based on 3-PBA toward beta-cypermethrin was consistent with that based on (trans+cis)-3-(2,2-dichlorovinyl)-2,2 dimethylcyclopropane carboxylic acid (DCCA). The analysis system is based on using liquid-liquid extraction and HPLC with a C18 column, gradient elution, and UV detection at 230 nm. The analytical methodology was validated by its repeatability, linearity, accuracy, and limits of detection. The developed and validated HPLC assay method was applied to examine the inhibition effect of S,S,S tributyl phosophorotrithioate (DEF) and piperonyl butoxide (PBO) on beta cypermethrin hydrolytic metabolism, and to evaluate the inducibility of six alpha cyano-3-phenoxybenzyl pyrethroids hydrolytic metabolism by 2-tridecanone, a generalized "inducer" of detoxifying enzyme activities. PMID- 20025276 TI - Extension of the QuEChERS method for pesticide residues in cereals to flaxseeds, peanuts, and doughs. AB - A simple method was evaluated for the determination of pesticide residues in flaxseeds, doughs, and peanuts using gas chromatography-time-of-flight mass spectrometry (GC-TOF) for analysis. A modified QuEChERS (quick, easy, cheap, effective, rugged, and safe) method, which was previously optimized for cereal grain samples, was evaluated in these fatty matrices. This extraction method involves first mixing the sample with 1:1 water/acetonitrile for an hour to swell the matrix and permit the salt-out liquid-liquid partitioning step using anhydrous MgSO(4) and NaCl. After shaking and centrifugation, cleanup is done by dispersive solid-phase extraction (d-SPE) using 150 mg of anhydrous MgSO(4), 150 mg of PSA, and 50 mg of C-18 per milliliter of extract. This method gave efficient separation of pesticides from fat and removal of coextracted substances better than gel permeation chromatography or use of a freeze-out step, which involved excessive use of solvent and/or time. The optimized analytical conditions were evaluated in terms of recoveries, reproducibilities, limits of detection, and matrix effects for 34 representative pesticides using different types of flaxseeds, peanuts, and doughs. Use of matrix-matched standards provided acceptable results for most pesticides with overall average recoveries between 70 and 120% and consistent RSDs <20% for semipolar pesticides and <26% for lipophilic pesticides. The recoveries of these latter types of pesticides depended on the fat content in the matrices and partitioning factor between the lipids and acetonitrile. We believe that the consistency of the pesticide recoveries for different samples in multiple experiments and the physicochemical partitioning explanation for <70% recoveries of lipophilic pesticides justify compensation of results for the empirically determined recovery values. In any case, this method still meets 10 ng/g detection limit needs for lipophilic pesticides and may be used for qualitative screening applications, in which any identified pesticides can be quantified and confirmed by a more intensive method that achieves >70% recoveries for lipophilic pesticides. PMID- 20025277 TI - Caseins: utilizing molecular chaperone properties to control protein aggregation in foods. AB - Denaturation and aggregation of proteins are reactions that are relevant to functional applications of proteins in foods. Depending on concentration, ionic strength, and pH, aggregation can result in turbidity, precipitation, or gelation. Aggregation may be desirable, as in the case of gelation, or undesirable, as in the case when it causes phase separation in beverages. One approach to improve the stability of globular proteins against heat stresses is through the addition of other compounds that alter aggregation. Numerous studies have shown the ability of molecular chaperones to assist proper folding/unfolding and assembly/disassembly of proteins, especially during stressed conditions. Recently, several papers have reported the molecular chaperone-like properties of caseins, especially using alpha(s)- and beta-caseins. Caseins appear to function like small heat shock proteins (sHSP). We have compared the results among investigations from the perspective of food processing conditions and related them to the mechanism for sHSP. Caseins possess three of the four common features among sHSP; lacking a similar sequence domain. Their function may be explained in part by having structures fitting the intrinsically unfolded class of proteins. With a few exceptions, most investigations were done at solution conditions that poorly represent foods; lacking investigations at pH < 4.5 and concentrations above 20 mg/mL. While it is clear that caseins can alter aggregation at neutral pH, their effectiveness at low pH, high protein concentration, and high thermal treatment (T >or= 100 degrees C) remains to be fully established. PMID- 20025278 TI - Newly mutated putative-aminotransferase in nonpungent pepper (Capsicum annuum) results in biosynthesis of capsinoids, capsaicinoid analogues. AB - Capsinoids make up a group of nonpungent capsaicinoid analogues produced in Capsicum fruits. They have bioactivities similar to those of capsaicinoids such as suppression of fat accumulation and antioxidant activity. Because of their low pungency, they are more palatable ingredients in dietary supplements than capsaicinoids. We recently reported that capsinoid biosynthesis is caused by nonsense mutation in a putative aminotransferase gene (p-AMT) in a nonpungent cultivar CH-19 Sweet. Here we report on the screening of nonpungent germplasm that revealed a nonpungent cultivar Himo, which contains high levels of capsinoids. We have shown that Himo has a recessive allele of p-amt, which contains a mutation different from that of CH-19 Sweet. Sequence analysis of p amt in Himo revealed that a single-nucleotide substitution results in one amino acid substitution from cysteine to arginine in the pyridoxal 5-phosphate binding domain. Genetic analysis using a cleaved amplified polymorphic sequence marker confirmed that the p-AMT genotype was precisely cosegregated with capsinoid biosynthesis and nonpungency. Himo will provide a new natural source of capsinoids. PMID- 20025279 TI - Anticancer effects of Alpinia pricei Hayata roots. AB - The leaves and roots of Alpinia pricei Hayata are used as a traditional wrapping for food and as a cooking substitute for fresh ginger. Our previous study showed that ethanol extracts from the roots of A. pricei Hayata (EEAP) and its phenolic compounds have anti-inflammatory effects. The aims of this work were to further study the in vitro anticancer activity of EEAP and its active compounds with respect to various cancer cells. The results from an MTT assay demonstrated that EEAP decreased the cell population growth of CH27, HL-60, and A549 cells. Flow cytometric analysis of HL-60 cells exposed to EEAP showed that the number of apoptotic cells increased in a time- and dose-dependent manner. Western blot data revealed that EEAP stimulated an increase in the level of protein expression of Fas, FasL, caspase-8, and tBid. Moreover, the ratio of the expression levels of pro- and anti-apoptotic Bcl-2 family members was changed after treatment with EEAP. EEAP-induced apoptosis involved the release of mitochondrial cytochrome c and subsequently induced the activation of caspase-9 and caspase-3, which were followed by the cleavage of poly(ADP-ribose) polymerase (PARP). The results also demonstrated that phenolic compounds (caffeic acid, apigenin, curcumin, and pinocembrin) from EEAP decreased the rate of population growth of HL-60 cells. Treatment of HL-60 cells with these phenolic compounds caused the loss of mitochondrial membrane potential. Our finding could provide critical information regarding the chemopreventive potential of ethanol extracts from A. pricei Hayata. These results also demonstrate that the EEAP-induced apoptotic ability in HL-60 cells might be related to the phenolic compounds. PMID- 20025280 TI - Probing Torpedo californica acetylcholinesterase catalytic gorge with two novel bis-functional galanthamine derivatives. AB - N-Piperidinopropyl-galanthamine (2) and N-saccharinohexyl-galanthamine (3) were used to investigate interaction sites along the active site gorge of Torpedo californica actylcholinesterase (TcAChE). The crystal structure of TcAChE-2 solved at 2.3 A showed that the N-piperidinopropyl group in 2 is not stretched along the gorge but is folded over the galanthamine moiety. This result was unexpected because the three carbon alkyl chain is just long enough for the bulky piperidine group to be placed above the bottleneck (Tyr121, Phe330) midway down the gorge. The crystal structure of TcAChE-3 at 2.2 A confirmed that a dual interaction with the sites at the bottom, and at the entrance of the gorge, enhances inhibitory activity: a chain of six carbon atoms has, in this class of derivatives, the correct length for optimal interactions with the peripheral anionic site (PAS). PMID- 20025282 TI - Evidence of increasing antibiotic resistance gene abundances in archived soils since 1940. AB - Mass production and use of antibiotics and antimicrobials in medicine and agriculture have existed for over 60 years, and has substantially benefited public health and agricultural productivity throughout the world. However, there is growing evidence that resistance to antibiotics (AR) is increasing both in benign and pathogenic bacteria, posing an emerging threat to public and environmental health in the future. Although evidence has existed for years from clinical data of increasing AR, almost no quantitative environmental data exist that span increased industrial antibiotic production in the 1950s to the present; i.e., data that might delineate trends in AR potentially valuable for epidemiological studies. To address this critical knowledge gap, we speculated that AR levels might be apparent in historic soil archives as evidenced by antibiotic resistance gene (ARG) abundances over time. Accordingly, DNA was extracted from five long-term soil-series from different locations in The Netherlands that spanned 1940 to 2008, and 16S rRNA gene and 18 ARG abundances from different major antibiotic classes were quantified. Results show that ARG from all classes of antibiotics tested have significantly increased since 1940, but especially within the tetracyclines, with some individual ARG being >15 times more abundant now than in the 1970s. This is noteworthy because waste management procedures have broadly improved and stricter rules on nontherapeutic antibiotic use in agriculture are being promulgated. Although these data are local to The Netherlands, they suggest basal environmental levels of ARG still might be increasing, which has implications to similar locations around the world. PMID- 20025281 TI - Using NMR solvent water relaxation to investigate metalloenzyme-ligand binding interactions. AB - This report demonstrates that solvent water relaxation measurements can be used for quantitative screening of ligand binding and for mechanistic investigations of enzymes containing paramagnetic metal centers by using conventional NMR instrumentation at high field. The method was exemplified using prolyl hydroxylase domain containing enzyme 2 (PHD2), a human enzyme involved in hypoxic sensing, with Mn(II) substituting for Fe(II) at the active site. K(D) values were determined for inhibitors that hinder access of water to the paramagnetic center. This technique is also useful for investigating the mechanism of suitable metalloenzymes, including order of ligand binding and modes of inhibition. PMID- 20025283 TI - Identification of structural properties associated with polychlorinated biphenyl dechlorination processes. AB - Polychlorinated biphenyl molecules can be biologically dechlorinated through sequential losses of a chlorine atom, following 840 pathways from higher chlorinated to lesser-chlorinated congeners and biphenyl. Previously, eight recurring sets of pathways, herein referred to as explicitly reported pathways in dechlorination processes, have been identified through qualitative analysis of shifts in congener masses in field and laboratory studies. Dechlorination process generalizations were qualitatively extrapolated based on limited attributes of the congeners dechlorinated in the explicitly reported pathways. They are valuable because they allow comparisons of dechlorination patterns across laboratory experiments and contaminated sites. However, due to analytical limitations and a paucity of studies, the explicitly reported pathways in dechlorination processes likely do not represent all of the pathways that could occur at contaminated sites. This work presents an alternative, quantitative, and replicable approach to the identification of candidate pathways for inclusion in dechlorination process generalizations through use of classification trees. This method considers 46 structural and property attributes of dechlorination pathways. Trees fit for pathway inclusion in each of the eight dechlorination processes with alternative assumptions are compared in terms of critical congener attributes. The classification trees correctly classify explicitly reported pathways into dechlorination processes at rates of 0.90 to 0.99. While many of the attributes used in the original generalizations were also selected as predictors by the classification trees, the extra attributes allow identification of additional dechlorination pathways that can be considered as candidates for monitoring in future studies. PMID- 20025284 TI - A semiempirical correlation between enthalpy of vaporization and saturation concentration for organic aerosol. AB - To model the temperature-induced partitioning of semivolatile organics in laboratory experiments or atmospheric models, one must know the appropriate heats of vaporization. Current treatments typically assume a constant value of the heat of vaporization or else use specific values from a small set of surrogate compounds. With published experimental vapor-pressure data from over 800 organic compounds, we have developed a semiempirical correlation between the saturation concentration (C*, microg m(-3)) and the heat of vaporization (deltaH(VAP), kJ mol(-1)) for organics in the volatility basis set. Near room temperature, deltaH(VAP) = -11 log(10)C(300)(*) + 129. Knowledge of the relationship between C* and deltaH(VAP) constrains a free parameter in thermodenuder data analysis. A thermodenuder model using our deltaH(VAP) values agrees well with thermal behavior observed in laboratory experiments. PMID- 20025285 TI - Compounds structurally related to Dechlorane Plus in sediment and biota from Lake Ontario (Canada). AB - The historical occurrence of Dechlorane Plus (DP) and detection of novel compounds structurally related to DP is described in a dated Lake Ontario sediment core. Our core was collected near the mouth of the Niagara River, which is known to be a major source of DP to the lake. Maximum DP concentrations (920 ng g(-1), dry weight) were observed between 1976 and 1980, the highest reported to date. Following that time, we observed a dramatic decrease in DP concentration which coincided with the enactment of United States federal and state laws to mitigate free release of chemicals into the Niagara River and installation of an industrial wastewater treatment facility. During the course of our research, four new substances structurally related to DP were also identified. These compounds were thought to arise from the Diels-Alder reactions resulting from impurities present in 1,5-cyclooctadiene, a feedstock used in production of DP. To confirm our hypothesis, Diels-Alder reactions were performed on the individual impurities. Using different stationary-phase capillary gas chromatography columns and high-resolution mass spectrometry, we were able to positively identify some of these novel compounds in the core. Interestingly, we also were able to identify a monoadduct compound, formed by addition of 1 mol of hexachlorocyclopentadiene to 2 mol of 1,3-cyclooctadiene, in lake trout. The concentration of this monoadduct was approximately 2 orders of magnitude greater than that of DP, suggesting that it is more bioaccumulative. PMID- 20025286 TI - Flame retardants are the suspected source of a new compound in the environment. PMID- 20025287 TI - pH-Dependent conformational changes in tear lipocalin by site-directed tryptophan fluorescence. AB - Tear lipocalin (TL), a major protein of human tears, binds a broad array of endogenous ligands. pH-dependent ligand binding in TL may have functional implications in tears. Previously, conformational selections of the AB and GH loops have been implicated in ligand binding by site-directed tryptophan fluorescence (SDTF). In this study, SDTF was applied to the AB and GH loops to investigate pH-driven conformational changes relevant to ligand binding. Both loops demonstrate significant but distinct conformational rearrangements over a wide pH range. In the low-pH transition, from 7.3 to 3.0, residues of the GH loop exhibit decreased solvent accessibilities. In acrylamide quenching experiments, the average quenching rate constant (k(q), accessibility parameter) of the residues in the GH loop is decreased approximately 38%, from 2.1 x 10(9) to 1.3 x 10(9) M(-1) s(-1). However, despite the significant changes in accessibilities for some residues in the AB loop, the average accessibility per residue remained unchanged (average k(q) = 1.2 M(-1) s(-1)). Accordingly, the low-pH transition induces conformational changes that reshuffle the accessibility profiles of the residues in the AB loop. A significant difference in the titration curves between the holo and apo forms of the W28 mutant suggests that the protonation states of the residues around position 28 modulate conformational switches of the AB loop relevant to ligand binding. PMID- 20025288 TI - Covalent bonding and the trans influence in lanthanide compounds. AB - A pair of mer-octahedral lanthanide chalcogenolate coordination complexes [(THF)(3)Ln(EC(6)F(5))(3) (Ln = Er, E = Se; Ln = Yb, E = S)] have been isolated and structurally characterized. Both compounds show geometry-dependent bond lengths, with the Ln-E bonds trans to the neutral donor tetrahydrofuran (THF) significantly shorter than the Ln-E bonds that are trans to negatively charged EC(6)F(5) ligands. Density functional theory calculations indicate that the structural trans influence evidenced by the differences in these bond lengths results from a covalent Ln-E interaction involving ligand p and Ln 5d orbitals. PMID- 20025289 TI - Pressure effects on a spin-crossover monomeric compound [Fe(pmea)(SCN)(2)] (pmea = bis[(2-pyridyl)methyl]-2-(2-pyridyl)ethylamine). AB - Spin-crossover (SCO) compounds are a sort of bistable material whose electronic and magnetic properties can be tuned by external physical stimuli, such as heat, light, and pressure. The title SCO compound [Fe(pmea)(NCS)(2)] (1; pmea = bis[(2 pyridyl)methyl]-2-(2-pyridyl)ethylamine) undergoes spin transition in such a way that it is an ideal candidate to investigate pressure effects on the SCO behavior. First, the spin transition is complete and abrupt so that the pressure dependent spin transition should be remarkable. Second, the T(1/2) value under ambient pressure is 184 K, which guarantees that the SCO temperature under various pressures does not exceed that restrained by high-pressure devices. The magnetic data of compound 1 under different external pressures were analyzed through a known method, as reported by Gutlich, which gave an interaction parameter Gamma of 264(5) cm(-1) and a volume change DeltaV degrees (HL) of 32(3) A(3) molecule(-1) [HL represents a high-spin (HS) <--> low-spin (LS) transition], respectively. Meanwhile, the calculated entropy change DeltaS degrees (HL)(T) at 1 bar is 59.79 J mol(-1) K(-1), which is a typical value that drives the spin transition from a LS to HS state. The pressure effects on the SCO behavior of compound 1 reported here may provide information for a deep understanding of the correlation between pressure and spin transition. PMID- 20025290 TI - Fully solution-processed inverted polymer solar cells with laminated nanowire electrodes. AB - We demonstrate organic photovoltaic cells in which every layer is deposited by solution processing on opaque metal substrates, with efficiencies similar to those obtained in conventional device structures on transparent substrates. The device architecture is enabled by solution-processed, laminated silver nanowire films serving as the top transparent anode. The cells are based on the regioregular poly(3-hexylthiophene) and C(61) butyric acid methyl ester bulk heterojunction and reach an efficiency of 2.5% under 100 mW/cm(2) of AM 1.5G illumination. The metal substrates are adequate barriers to moisture and oxygen, in contrast to transparent plastics that have previously been used, giving rise to the possibility of roll-to-roll solution-processed solar cells that are packaged by lamination to glass substrates, combining the cost advantage of roll to-roll processing with the barrier properties of glass and metal foil. PMID- 20025292 TI - Design and synthesis of novel diaminoquinazolines with in vivo efficacy for beta catenin/T-cell transcriptional factor 4 pathway inhibition. AB - We are introducing a novel series of 2,4-diaminoquinazolines as beta-catenin/Tcf4 inhibitors which were identified by ligand-based design. Here we elucidate the SAR of this series and explain how we were able to improve key molecular properties such as solubility and cLogP leading to compound 9. Analogue 9 exhibited better biological activity and improved physical and pharmacological properties relative to the HTS hit 49. Furthermore, 9 demonstrated good cell growth inhibition against several human colorectal cancer lines such as LoVo and HT29. In addition, treatment with compound 9 led to gene expression changes that overlapped significantly with the transcriptional profile resulting from the pathway inhibition by siRNA knockdown of beta-catenin or Tcf4. Subsequently, 9 was tested for efficacy in a beta-catenin/RKE-mouse xenograft, where it led to more then 50% decrease in tumor volume. PMID- 20025291 TI - Expanding the family of collagen proteins: recombinant bacterial collagens of varying composition form triple-helices of similar stability. AB - The presence of the (Gly-Xaa-Yaa)(n) open reading frames in different bacteria predicts the existence of an expanded family of collagen-like proteins. To further explore the triple-helix motif and stabilization mechanisms in the absence of hydroxyproline (Hyp), predicted novel collagen-like proteins from Gram positive and -negative bacteria were expressed in Escherichia coli and characterized. Soluble proteins capable of successful folding and in vitro refolding were observed for collagen proteins from Methylobacterium sp 4-46, Rhodopseudomonas palustris and Solibacter usitatus . In contrast, all protein constructs from Clostridium perfringens were found predominantly in inclusion bodies. However, attachment of a heterologous N-terminal or C-terminal noncollagenous folding domain induced the Clostridium perfringens collagen domain to fold and become soluble. The soluble constructs from different bacteria had typical collagen triple-helical features and showed surprisingly similar thermal stabilities despite diverse amino acid compositions. These collagen-like proteins provide a resource for the development of biomaterials with new properties. PMID- 20025293 TI - Clickable, biocompatible, and fluorescent hybrid nanoparticles for intracellular delivery and optical imaging. AB - We report a general and facile approach for the fabrication of a new class of near monodisperse hybrid nanoparticles via RAFT polymerization and self-assembly in water. Furthermore, we combine a fluorescent inorganic silica core with a biocompatible polymer shell and a terminal unit susceptible to facile conjugations via click chemistry. A tailoring of the weight fractions of both components allows a tuning of the size of the formed aggregates. Fluorescent properties and the crosslinking into an organic-inorganic hybrid network are realized by copolymerizing a dye-functionalized monomer 1-pyrenebutyl acrylate and a trimethoxysilane-carrying one, (3-acryloxypropyl)trimethoxysilane. The potential of these stabilized and fluorescent nanoparticles as biocompatible carriers for intracellular delivery is demonstrated via in vitro experiments on lung cancer cells. PMID- 20025294 TI - ApoE induces serum paraoxonase PON1 activity and stability similar to ApoA-I. AB - Serum paraoxonase (PON1) is an anti-atherogenic interfacially activated lipo lactonase that was shown to selectively bind high-density lipoprotein (HDL) carrying apolipoprotein A-I (apoA-I). ApoA-I binding occurs with nanomolar affinity and induces a dramatic increase in enzyme stability and lactonase activity. This study examined the association of PON1 with reconstituted HDL (rHDL) carrying apolipoprotein E, and its consequences on the stability and enzymatic activity of PON1, and on its anti-atherogenic potential. The results indicate that reconstituted HDL particles prepared with two most common isoforms of apoE (apoE3 and apoE4) associate with rePON1 in a manner and affinity similar to those of apoA-I. Binding to apoE-HDL stimulates the lactonase activity and stabilizes the enzyme, although the latter occurs to a >10-fold lesser extent compared to apoA-I-HDL particles. The anti-atherogenic potential of PON1, measured by inhibition of LDL oxidation and stimulation of macrophage cholesterol efflux, was also stimulated by apoE-HDL, at levels of 40-96% compared to apoA-I HDL. Overall, reconstituted apoE-HDL exhibits properties similar to those of apoA I-HDL, but with a lower capacity to stabilize PON1 and to induce its anti atherogenic functions. ApoE, apoA-I, and to a lesser degree apoA-IV show distinct structural and functional similarities but little sequence homology. That these apolipoproteins, but not apoA-II, bind PON1 with high affinity and stimulate its activity suggests that PON1-HDL recognition is based primarily on surface properties of the apolipoproteins and that specific protein-protein interactions may play only a secondary role. PMID- 20025295 TI - The mechanism of polyplex internalization into cells: testing the GM1/caveolin-1 lipid raft mediated endocytosis pathway. AB - The GM1/caveolin-1 lipid raft mediated endocytosis mechanism was explored for generation 5 and 7 poly(amidoamine) dendrimer polyplexes employing the Cos-7, 293A, C6, HeLa, KB, and HepG2 cell lines. Expression levels of GM1 and caveolin-1 were measured using dot blot and Western blot, respectively. The level of GM1 in the cell plasma membrane was adjusted by incubation with exogenous GM1 or ganglioside inhibitor PPMP, and the level of CAV-1 was adjusted by upregulation with the adenovirus vector expressed caveolin-1 (AdCav-1). Cholera toxin B subunit was employed as a positive control for uptake in all cases. No evidence was found for a GM1/caveolin-1 lipid raft mediated endocytosis mechanism for the generation 5 and 7 poly(amidoamine) dendrimer polyplexes. PMID- 20025296 TI - Mass spectral analysis of neuropeptide expression and distribution in the nervous system of the lobster Homarus americanus. AB - The lobster Homarus americanus has long served as an important animal model for electrophysiological and behavioral studies. Using this model, we performed a comprehensive investigation of the neuropeptide expression and their localization in the nervous system, which provides useful insights for further understanding of their biological functions. Using nanoLC ESI Q-TOF MS/MS and three types of MALDI instruments, we analyzed the neuropeptide complements in a major neuroendocrine structure, pericardial organ. A total of 57 putative neuropeptides were identified and 18 of them were de novo sequenced. Using direct tissue/extract analysis and bioinformatics software SpecPlot, we charted the global distribution of neuropeptides throughout the nervous system in H. americanus. Furthermore, we also mapped the localization of several neuropeptide families in the brain by high mass resolution and high mass accuracy mass spectrometric imaging (MSI) using a MALDI LTQ Orbitrap mass spectrometer. We have also compared the utility and instrument performance of multiple mass spectrometers for neuropeptide analysis in terms of peptidome coverage, sensitivity, mass spectral resolution and capability for de novo sequencing. PMID- 20025299 TI - Carbon nanotubes and microwaves: interactions, responses, and applications. AB - The interaction of microwaves with carbon nanotubes (CNTs) is an interesting topic for a variety of potential applications. Microwaves have been used for the purification of CNTs and for their chemical functionalization, providing a technique for simple, green, and large-scale protocols. In addition, the selective destruction of metallic CNTs under microwave irradiation could potentially result in a batch of semiconducting-only nanotubes. As an innovative application, the combination of microwaves with well-aligned CNTs could produce a new illumination technology. Moreover, the microwave absorbing properties of CNTs and their different behavior from typical organic compounds may open the door to the preparation of a wide range of new materials useful in many fields. A few examples of practical applications include electromagnetic interference for protecting the environment from radiation and microwave hyperthermia for cancer treatment as well as other medical therapies requiring precise heating of biological tissues. PMID- 20025300 TI - Beyond the sparkle: the impact of nanodiamonds as biolabeling and therapeutic agents. AB - A paper by Treussart and co-workers in this issue demonstrates the application of photoluminescent nanodiamonds for intracellular labeling as well as mechanistic cellular uptake studies. Findings from this paper reveal that optimal photoluminescence of nitrogen-vacancy color centers can be attained with photostability and no photoblinking, enabling continuous tracking in the cytoplasm over sustained time scales. In addition to the fluorescent properties of the nanodiamonds, internalization assays reveal a primarily endocytic uptake process. A high degree of nanodiamond ( approximately 46 nm in diameter) and endosome colocalization as well as cytoplasmic presence of smaller nanodiamonds was observed. Several attributes of the nanodiamond particles are elucidated in this and other recent studies, ranging from their stability as imaging agents to their potential as intracellular molecular delivery vehicles. These findings give insight into the use of nanodiamonds as an emerging platform for therapeutic and diagnostic ("theranostic") nanomedicine, forging new foundations and criteria for continued nanodiamond engineering toward downstream clinical relevance and impact. PMID- 20025301 TI - Assessing the safety of nanomaterials by genomic approach could be another alternative. PMID- 20025303 TI - Origin of laser-induced near-subwavelength ripples: interference between surface plasmons and incident laser. AB - We show that short-pulse laser-induced classical ripples on dielectrics, semiconductors, and conductors exhibit a prominent "non-classical" characteristic in normal incidence the periods are definitely smaller than laser wavelengths, which indicates that the simplified scattering model should be revised. Taking into account the surface plasmons (SPs), we consider that the ripples result from the initial direct SP-laser interference and the subsequent grating-assisted SP laser coupling. With the model, the period-decreasing phenomenon originates in the admixture of the field-distribution effect and the grating-coupling effect. Further, we propose an approach for obtaining the dielectric constant, electron density, and electron collision time of the high-excited surface. With the derived parameters, the numerical simulations are in good agreement with the experimental results. On the other hand, our results confirm that the surface irradiated by short-pulse laser with damage-threshold fluence should behave metallic, no matter for metal, semiconductor, or dielectric, and the short-pulse laser-induced subwavelength structures should be ascribed to a phenomenon of nano optics. PMID- 20025304 TI - Nanometer-scale striped surface terminations on fractured SrTiO(3) surfaces. AB - Using cross-sectional scanning tunneling microscopy on in situ fractured SrTiO(3), one of the most commonly used substrates for the growth of complex oxide thin films and superlattices, atomically smooth terraces have been observed on (001) surfaces. Furthermore, it was discovered that fracturing this material at room temperature results in the formation of stripe patterned domains having characteristic widths ( approximately 10 to approximately 20 nm) of alternating surface terminations that extend over a long range. Spatial characterization utilizing spectroscopy techniques revealed a strong contrast in the electronic structure of the two domains. Combining these results with topographic data, we are able to assign both TiO(2) and SrO terminations to their respective domains. The results of this experiment reveal that fracturing this material leads to reproducibly flat surfaces that can be characterized at the atomic-scale and suggest that this technique can be utilized for the study of technologically relevant complex oxide interfaces. PMID- 20025305 TI - Koopmans' springs to life. AB - The meaning of orbital energies (OOEs) in Kohn-Sham (KS) density functional theory (DFT) is subject to a longstanding controversy. In local, semilocal, and hybrid density functionals (DFs) a Koopmans' approach, where OOEs approximate negative ionization potentials (IPs), is unreliable. We discuss a methodology based on the Baer-Neuhauser-Livshits range-separated hybrid DFs for which Koopmans' approach "springs to life." The OOEs are remarkably close to the negative IPs with typical deviances of +/-0.3 eV down to IPs of 30 eV, as demonstrated on several molecules. An essential component is the ab initio motivated range-parameter tuning procedure, forcing the highest OOE to be exactly equal to the negative first IP. We develop a theory for the curvature of the energy as a function of fractional occupation numbers to explain some of the results. PMID- 20025306 TI - Computing absolute free energies of disordered structures by molecular simulation. AB - We present a Monte Carlo simulation technique by which the free energy of disordered systems can be computed directly. It is based on thermodynamic integration. The central idea is to construct an analytically solvable reference system from a configuration which is representative for the state of interest. The method can be applied to lattice models (e.g., the Ising model) as well as off-lattice molecular models. We focus mainly on the more challenging off-lattice case. We propose a Monte Carlo algorithm, by which the thermodynamic integration path can be sampled efficiently. At the examples of the hard sphere liquid and a hard disk solid with a defect, we discuss several properties of the approach. PMID- 20025307 TI - Single chain contraction and re-expansion of polystyrene sulfonate: a study on its re-entrant condensation at single molecular level. AB - Single chain conformation of a polyelectrolyte (polystyrene sulfonate, PSS( )Na(+)) during its re-entrant condensation was studied by fluorescence correlation spectroscopy (FCS) with single molecule sensitivity. The contraction and re-expansion of PSS(-)Na(+) chain were observed with the addition of counterions of different valencies. The formation of aggregation and precipitation of PSS(-)Na(+) and its redissolution were observed in accordance with the chain contraction and re-expansion process for the PSS(-)Na(+) chain upon the addition of trivalent La(3+) ion. Chain contraction and re-expansion of the PSS(-)Na(+) chain were also observed with the addition of monovalent Cs(+) and divalent Ca(2+) ions, under which condition, the re-entrant condensation was not observed. The results demonstrate that the high sensitivity of FCS can really study single PSS(-)Na(+) chain under extremely dilute situation. PMID- 20025308 TI - Towards an accurate description of the electronic properties of the biphenylthiol/gold interface: the role of exact exchange. AB - We investigate the role of the exact exchange in describing the biphenylthiol/gold interface. The study is performed by simulating the electronic properties of mercaptobiphenylthiol and aminobiphenylthiol molecules adsorbed on a Au(23) cluster, using local, semilocal and hybrid functionals and an effective exact exchange method, namely, the localized Hartree-Fock (LHF). We find that the local/semilocal functionals strongly underestimate the charge transfer and the bond dipole at the interface due to the self-interaction-error (SIE), which alters the correct level alignment. On the other hand the LHF method is SIE free and predicts a larger charge transfer and bond dipole. We also found that LHF results can be reproduced using hybrid functionals and that conventional local/semilocal correlation functionals are unable to improve over the exchange only description. PMID- 20025309 TI - New shooting algorithms for transition path sampling: centering moves and varied perturbation sizes for improved sampling. AB - Transition path sampling has been established as a powerful tool for studying the dynamics of rare events. The trajectory generation moves of this Monte Carlo procedure, shooting moves and shifting modes, were developed primarily for rate constant calculations, although this method has been more extensively used to study the dynamics of reactive processes. We have devised and implemented three alternative trajectory generation moves for use with transition path sampling. The centering-shooting move incorporates a shifting move into a shooting move, which centers the transition period in the middle of the trajectory, eliminating the need for shifting moves and generating an ensemble where the transition event consistently occurs near the middle of the trajectory. We have also developed varied-perturbation size shooting moves, wherein smaller perturbations are made if the shooting point is far from the transition event. The trajectories generated using these moves decorrelate significantly faster than with conventional, constant sized perturbations. This results in an increase in the statistical efficiency by a factor of 2.5-5 when compared to the conventional shooting algorithm. On the other hand, the new algorithm breaks detailed balance and introduces a small bias in the transition time distribution. We have developed a modification of this varied-perturbation size shooting algorithm that preserves detailed balance, albeit at the cost of decreased sampling efficiency. Both varied-perturbation size shooting algorithms are found to have improved sampling efficiency when compared to the original constant perturbation size shooting algorithm. PMID- 20025310 TI - Optimizing core-shell nanoparticle catalysts with a genetic algorithm. AB - A genetic algorithm is used with density functional theory to investigate the catalytic properties of 38- and 79-atom bimetallic core-shell nanoparticles for the oxygen reduction reaction. Each particle is represented by a two-gene chromosome that identifies its core and shell metals. The fitness of each particle is specified by how close the d-band level of the shell is to that of the Pt(111) surface, a catalyst known to be effective for oxygen reduction. The genetic algorithm starts by creating an initial population of random core-shell particles. The fittest particles are then bred and mutated to replace the least fit particles in the population and form successive generations. The genetic algorithm iteratively refines the population of candidate catalysts more efficiently than Monte Carlo or random sampling, and we demonstrate how the average energy of the surface d-band can be tuned to that of Pt(111) by varying the core and shell metals. The binding of oxygen is a more direct measure of catalytic activity and is used to further investigate the fittest particles found by the genetic algorithm. The oxygen binding energy is found to vary linearly with the d-band level for particles with the same shell metal, but there is considerable variation in the trend across different shells. Several particles with oxygen binding energies similar to Pt(111) have already been investigated experimentally and found to be active for oxygen reduction. In this work, many other candidates are identified. PMID- 20025311 TI - Analytical formulas for low-fluence non-line-narrowed hole-burned spectra in an excitonically coupled dimer. AB - We present exact equations for the low-fluence non-line-narrowed (NLN) nonphotochemical hole-burning (NPHB) spectrum of an excitonically coupled dimer (for arbitrary coupling strength) under the assumption that postburn and preburn site energies are independent. The equations provide a transparent view into the contributions of various effects to the NPHB spectrum. It is demonstrated that the NPHB spectrum in dimers is largely dominated by the statistical reshuffling of site energies and by altered excitonic transition energies of both excitonic states (in contrast with only the lowest state). For comparison of these results with those from larger excitonically coupled systems, the low-fluence NLN NPHB spectrum obtained for the CP47 complex (a 16-pigment core antenna complex of Photosystem II) is also calculated using Monte Carlo simulations. In this larger system it is shown that the NPHB spectra for individual excitonic states are not entirely conservative (although the changes in average oscillator strength for the higher excitonic states are in most cases less than 1%), a feature which we argue is due primarily to reordering of the contributions of various pigments to the excitonic states. We anticipate that a better understanding of NPHB spectra obtained for various photosynthetic complexes and their simultaneous fits with other optical spectra (e.g., absorption, emission, and circular dichroism spectra) will provide more insight into the underlying electronic structures of various photosynthetic systems. PMID- 20025312 TI - Two-scale Brownian dynamics of suspensions of charged nanoparticles including electrostatic and hydrodynamic interactions. AB - We propose here a multiscale strategy based on continuous solvent Brownian dynamics (BD) simulations to study the dynamical properties of aqueous suspensions of charged nanoparticles. We extend our previous coarse-graining strategy [V. Dahirel et al., J. Chem. Phys. 126, 114108 (2007)] to account for hydrodynamic interactions between solute particles. Within this new procedure, two BD simulations are performed: (1) The first one investigates the time scales of the counterions and coions (the microions) with only one nanoparticle in the simulation box but explicit microions, (ii) the second one investigates the larger time scale of the nanoparticles with numerous nanoparticles in the simulation box but implicit microions. We show how individual and collective transport coefficients can be computed from this two-scale procedure. To ensure the validity of our procedure, we compute the transport coefficients of a 10-1 model electrolyte in aqueous solution with a 1-1 added salt. We do a systematic comparison between the results obtained within the new procedure and those obtained with explicit BD simulations of the complete system containing several nanoparticles and explicit microions. The agreement between the two methods is found to be excellent: Even if the new procedure is much faster than explicit simulations, it allows us to compute transport coefficients with a good precision. Moreover, one step of our procedure also allows us to compute the individual transport coefficients relative to the microions (self-diffusion coefficients and electrophoretic mobility). PMID- 20025313 TI - Nonlinear diffusion in two-dimensional ordered porous media based on a free volume theory. AB - A continuum nonlinear diffusion model is developed to describe molecular transport in ordered porous media. An existing generic van der Waals equation of state based free volume theory of binary diffusion coefficients is modified and introduced into the two-dimensional diffusion equation. The resulting diffusion equation is solved numerically with the alternating-direction fully implicit method under Neumann boundary conditions. Two types of pore structure symmetries are considered, hexagonal and cubic. The former is modeled as parallel channels while in case of the latter equal-sized channels are placed perpendicularly thus creating an interconnected network. First, general features of transport in both systems are explored, followed by the analysis of the impact of molecular properties on diffusion inside and out of the porous matrix. The influence of pore size on the diffusion-controlled release kinetics is assessed and the findings used to comment recent experimental studies of drug release profiles from ordered mesoporous silicates. PMID- 20025314 TI - Extensive regularization of the coupled cluster methods based on the generating functional formalism: application to gas-phase benchmarks and to the SN2 reaction of CHCl3 and OH- in water. AB - The recently introduced energy expansion based on the use of generating functional (GF) [K. Kowalski and P. D. Fan, J. Chem. Phys. 130, 084112 (2009)] provides a way of constructing size-consistent noniterative coupled cluster (CC) corrections in terms of moments of the CC equations. To take advantage of this expansion in a strongly interacting regime, the regularization of the cluster amplitudes is required in order to counteract the effect of excessive growth of the norm of the CC wave function. Although proven to be efficient, the previously discussed form of the regularization does not lead to rigorously size-consistent corrections. In this paper we address the issue of size-consistent regularization of the GF expansion by redefining the equations for the cluster amplitudes. The performance and basic features of proposed methodology are illustrated on several gas-phase benchmark systems. Moreover, the regularized GF approaches are combined with quantum mechanical molecular mechanics module and applied to describe the S(N)2 reaction of CHCl(3) and OH(-) in aqueous solution. PMID- 20025315 TI - Derivatives of spin dynamics simulations. AB - We report analytical equations for the derivatives of spin dynamics simulations with respect to pulse sequence and spin system parameters. The methods described are significantly faster, more accurate, and more reliable than the finite difference approximations typically employed. The resulting derivatives may be used in fitting, optimization, performance evaluation, and stability analysis of spin dynamics simulations and experiments. PMID- 20025316 TI - Automatic derivation and evaluation of vibrational coupled cluster theory equations. AB - A scheme for automatic derivation and evaluation of the expressions occurring in vibrational coupled cluster theory is introduced. The method is based on a Baker Campbell-Hausdorff expansion of the similarity transformed Hamiltonian and is general both with respect to the excitation level in the parameter space and the mode coupling level in the Hamiltonian. In addition to deriving general expressions, intermediates that lower the computational scaling are automatically detected. The final equations are then evaluated. Due to the commutator based nature of the algorithm, it is also applicable to the evaluation of quantities needed for response theory. Different aspects of the theory and implementation are illustrated by calculations on model systems. Furthermore, all fundamental excitation energies of ethylene oxide are calculated. PMID- 20025317 TI - A density-division embedding potential inversion technique. AB - A new method is proposed to partition the density of a system in two portions. The density on each subsystem is the solution of a Fock equation modified by the addition of an embedding potential. This embedding potential is obtained iteratively by minimizing the difference between the electronic densities of the total system and the sum of the subsystems. Thus, the electronic density partition and the embedding potential are obtained at the same time within the procedure, guaranteeing the v-representability of the densities partitioned. This fact is a considerable improvement of a recently proposed embedding potential inversion technique, [O. Roncero, M. P. de Lara-Castells, P. Villarreal, F. Flores, J. Ortega, M. Paniagua, and A. Aguado, J. Chem. Phys. 129, 184104 (2008)], in which the embedding potential is obtained once the electronic density is previously partitioned. The method is first applied to a linear H(10) chain to illustrate how it works. The orbitals obtained are localized on each subsystem, and can be used to include local electronic correlation with currently available ab initio programs. Finally, the method is applied to include the electronic correlation needed to describe the van der Waals interaction between H(10) chains and H(2) molecules, of approximately 12 meV, giving very accurate results. PMID- 20025318 TI - A simple nonlocal model for exchange. AB - This work presents a new nonlocal model for the exchange energy density. The model is obtained from the product of the Kohn-Sham one-particle density matrix used to construct exact [Hartree-Fock-like (HF)] exchange, and an approximate density matrix used to construct local spin-density approximation (LSDA) exchange. The proposed exchange energy density has useful formal properties, including correct spin and coordinate scaling and the correct uniform limit. It can readily be evaluated in finite basis sets, with a computational scaling intermediate between HF exchange and semilocal quantities such as the noninteracting kinetic energy density. Applications to representative systems indicate that its properties are typically intermediate between HF and LSDA exchange, and often similar to global hybrids of HF and LSDA exchange. The model is proposed as a novel "Rung 3.5" ingredient for constructing approximate exchange-correlation functionals. PMID- 20025319 TI - Isotope shift in the electron affinity of lithium. AB - Very accurate electron affinity (EA) calculations of (6)Li and (7)Li (and (infinity)Li) have been performed using explicitly correlated Gaussian functions and a variational approach that explicitly includes the nuclear motion in the calculations (i.e., the approach that does not assume the Born-Oppenheimer approximation). The leading relativistic and quantum electrodynamics corrections to the electron affinities were also calculated. The results are the most accurate theoretical values obtained for the studied systems to date. Our best estimates of the (7)Li and (6)Li EAs are 4984.9842(30) and 4984.9015(30) cm(-1), respectively, and of the (7)Li/(6)Li EA isotope shift is 0.0827 cm(-1). PMID- 20025320 TI - Thermal conduction in molecular materials using coarse grain dynamics: role of mass diffusion and quantum corrections for molecular dynamics simulations. AB - We use a mesodynamical method, denoted dynamics with implicit degrees of freedom (DID), to characterize thermal transport in a model molecular crystal below and above its melting temperature. DID represents groups of atoms (molecules in this case) using mesoparticles and the thermal role of the intramolecular degrees of freedom (DoFs) are described implicitly using their specific heat. We focus on the role of these intramolecular DoFs on thermal transport. We find that thermal conductivity is independent of intramolecular specific heat for solid samples and a linear relationship between the two quantities in liquid samples with the coefficient of proportionality being the mass diffusivity of the mesoparticles. As the temperature of the liquids is increased, thermal conductivity exhibits an increased sensitivity with respect to the specific heat of the internal DoFs due to the enhanced molecular mobility. Based on these results, we propose a simple method to incorporate quantum corrections to thermal conductivity obtained from nonequilibrium molecular dynamics simulations of molecular liquids. Our results also provide insight into the development of thermally accurate coarse grain models of soft materials. PMID- 20025321 TI - Continuous-time random walks at all times. AB - Continuous-time random walks (CTRW) play an important role in understanding of a wide range of phenomena. However, most theoretical studies of these models concentrate only on dynamics at long times. We present a new theoretical approach, based on generalized master equations picture, which allowed us to obtain explicit expressions for Laplace transforms for all dynamic quantities for different CTRW models. This theoretical method leads to the effective description of CTRW at all times. Specific calculations are performed for homogeneous, periodic models and for CTRW with irreversible detachments. The approach to stationary states for CTRW is analyzed. Our results are also used to analyze generalized fluctuations theorem. PMID- 20025322 TI - Modeling the nanoscale viscoelasticity of fluids by bridging non-Markovian fluctuating hydrodynamics and molecular dynamics simulations. AB - A multiscale computational method is developed to model the nanoscale viscoelasticity of fluids by bridging non-Markovian fluctuating hydrodynamics (FHD) and molecular dynamics (MD) simulations. To capture the elastic responses that emerge at small length scales, we attach an additional rheological model parallel to the macroscopic constitutive equation of a fluid. The widely used linear Maxwell model is employed as a working choice; other models can be used as well. For a fluid that is Newtonian in the macroscopic limit, this approach results in a parallel Newtonian-Maxwell model. For water, argon, and an ionic liquid, the power spectrum of momentum field autocorrelation functions of the parallel Newtonian-Maxwell model agrees very well with those calculated from all atom MD simulations. To incorporate thermal fluctuations, we generalize the equations of FHD to work with non-Markovian rheological models and colored noise. The fluctuating stress tensor (white noise) is integrated in time in the same manner as its dissipative counterpart and numerical simulations indicate that this approach accurately preserves the set temperature in a FHD simulation. By mapping position and velocity vectors in the molecular representation onto field variables, we bridge the non-Markovian FHD with atomistic MD simulations. Through this mapping, we quantitatively determine the transport coefficients of the parallel Newtonian-Maxwell model for water and argon from all-atom MD simulations. For both fluids, a significant enhancement in elastic responses is observed as the wave number of hydrodynamic modes is reduced to a few nanometers. The mapping from particle to field representations and the perturbative strategy of developing constitutive equations provide a useful framework for modeling the nanoscale viscoelasticity of fluids. PMID- 20025323 TI - Theoretical investigation of the ground and low-lying excited states of gallium and indium silicides, GaSi and InSi. AB - The electronic structure and the bonding of 13 electronic states of the diatomic gallium silicide (GaSi) and indium silicide (InSi) have been studied by multireference configuration interaction and coupled clusters methods in conjunction with the basis set (aug-)cc-pwCVQZ(-PP). Potential energy curves have been constructed for all states. Binding energies, spectroscopic parameters, and dipole moments have been calculated. Moreover, the bonding in the different states is analyzed. The potential energy curves of the two molecules, GaSi and InSi, are similar. The binding energies and the bond distances of the ground states of the molecules, X (4)Sigma(-), are D(e)=56.2 kcal/mol (GaSi) and 51.9 kcal/mol (InSi) and R(e)=2.406 A (GaSi) and 2.603 A (InSi). The first excited state a (2)Sigma(-) is calculated at 17 kcal/mol above the ground state in both molecules. PMID- 20025324 TI - Long-range collisional energy transfer between charge-transfer (ion-pair) states of I2, induced by H2O and I2(X). AB - Long-range (resonant) energy transfer, between g/u charge-transfer states of molecular iodine [i.e., f0(g) (+)((3)P(0))-->F0(u)(+)((3)P(0)) and E0(g)(+)((3)P(2))-->D0(u) (+)((3)P(2))], induced by collisions with H(2)O and I(2)(X) via multipole coupling, has been observed. Large rate constants, up to 5 x 10(-9) molecules(-1) cm(3) s(-1), for collisional transfer between a range of vibrational levels of the f0(g)(+)((3)P(0)) and F0(u)(+)((3)P(0)) ion-pair states of I(2), by H(2)O, are reported. Some previously reported studies on E0(g)(+)((3)P(2))-->D0(u)(+)((3)P(2)) and f0(g)(+)((3)P(0))-->F0(u)(+)((3)P(0)) collisional transfer, induced by I(2)(X), have been repeated and revised rate data are presented; the range of initially excited vibrational states studied has also been extended. Much smaller rate constants for quenching by I(2)(X), compared to H(2)O, are found and it is proposed that H(2)O desorbed from the walls of the sample cell could have significantly affected much larger rate data previously reported in the literature. For both collision partners, a model is proposed in which long-range, near-resonant interactions can occur when there is close matching of the change in energy in the ion-pair states with the change in energy that accompanies the rotational transition undergone by the collision partner. PMID- 20025325 TI - A single molecule as a dielectric medium. AB - For three molecules with weak or negligible charge overlap, we prove that the three-body interaction energy obtained from quantum perturbation theory (to leading order) fits a dielectric model with a nonlocal electronic screening function. The electronic charge cloud of each molecule acts as a dielectric medium for the interaction of the remaining two with the nonlocal dielectric function epsilon(r,r') obtained by O. S. Jenkins and K. L. C. Hunt [J. Chem. Phys. 119, 8250 (2003)], by considering the charge redistribution induced in a single molecule by an external perturbation. The dielectric function depends parametrically on the coordinates of the nuclei, within the Born-Oppenheimer approximation. We also prove that the force on each nucleus in molecule A depends on intramolecular dielectric screening within A. The potential from the charge distribution of B, screened by C acting as a dielectric medium, is further screened linearly within A; and similarly, with the roles of B and C reversed. In addition, the potential due to the unperturbed charge distribution of B and the potential due to the unperturbed charge distribution of C, acting simultaneously, are screened nonlinearly within A. The results show that nonlocal dielectric theory holds on the molecular level, provided that the overlap of the electronic charge distributions is weak. PMID- 20025326 TI - A new experimental absolute nuclear magnetic shielding scale for oxygen based on the rotational hyperfine structure of H(2)(17)O. AB - The hyperfine structure in the rotational spectrum of water containing (17)O has been investigated experimentally and by means of quantum-chemical calculations. The Lamb-dip technique has been used to resolve the hyperfine structure due to spin-rotation as well as spin-spin interactions and allowed the determination of the corresponding hyperfine parameters with high accuracy. The experimental investigation and, in particular, the analysis of the spectra have been supported by quantum-chemical computations at the coupled-cluster level. The experimental (17)O isotropic spin-rotation constant of H(2)(17)O has been used in a further step for the determination of the paramagnetic part of the corresponding nuclear magnetic shielding constant, whereas the diamagnetic contribution as well as vibrational and temperature corrections have been obtained from quantum-chemical calculations. This joint procedure leads to a value of 325.3(3) ppm for the oxygen shielding in H(2)(17)O at 300 K, in good agreement with pure theoretical predictions, and in this way provides the basis for a new absolute oxygen shielding scale. PMID- 20025327 TI - CO chemisorption on the surfaces of the golden cages. AB - We report a joint experimental and theoretical study of CO chemisorption on the golden cages. We find that the Au(17)(-) cage is highly robust and retains its cage structure in Au(17)(CO)(-). On the other hand, the Au(16)(-) cage is transformed to a structure similar to Au(17)(-) upon the adsorption of CO. Au(18)(-) is known to consist of two nearly degenerate structures, i.e., a cage and a pyramidal isomer, which coexist in the cluster beam. However, upon CO chemisorption only the cage isomer is observed while the pyramidal isomer no longer exists due to its less favorable interaction with CO, compared to the cage isomer. We find that inclusion of the spin-orbit effects is critical in yielding simulated spectra in quantitative agreement with the experimental data and providing unequivocal structural information and molecular insights into the chemical interactions between CO and the golden cages. PMID- 20025328 TI - Conical intersections, charge localization, and photoisomerization pathway selection in a minimal model of a degenerate monomethine dye. AB - We propose a minimal model Hamiltonian for the electronic structure of a monomethine dye, in order to describe the photoisomerization of such dyes. The model describes interactions between three diabatic electronic states, each of which can be associated with a valence bond structure. Monomethine dyes are characterized by a charge-transfer resonance; the indeterminacy of the single double bonding structure dictated by the resonance is reflected in a duality of photoisomerization pathways corresponding to the different methine bonds. The possible multiplicity of decay channels complicates mechanistic models of the effect of the environment on fluorescent quantum yields, as well as coherent control strategies. We examine the extent and topology of intersection seams between the electronic states of the dye and how they relate to charge localization and selection between different decay pathways. We find that intersections between the S(1) and S(0) surfaces only occur for large twist angles. In contrast, S(2)/S(1) intersections can occur near the Franck-Condon region. When the molecule has left-right symmetry, all intersections are associated with con- or disrotations and never with single bond twists. For asymmetric molecules (i.e., where the bridge couples more strongly to one end) the S(2) and S(1) surfaces bias torsion about different bonds. Charge localization and torsion pathway biasing are correlated. We relate our observations with several recent experimental and theoretical results, which have been obtained for dyes with similar structure. PMID- 20025329 TI - An integrated approach to NMR spin relaxation in flexible biomolecules: application to beta-D-glucopyranosyl-(1-->6)-alpha-D-mannopyranosyl-OMe. AB - The description of the reorientational dynamics of flexible molecules is a challenging task, in particular when the rates of internal and global motions are comparable. The commonly used simple mode-decoupling models are based on the assumption of statistical independence between these motions. This assumption is not valid when the time scale separation between their rates is small, a situation that was found to arise in oligosaccharides in the context of certain internal motions. To make possible the interpretation of NMR spin relaxation data from such molecules, we developed a comprehensive approach generally applicable to flexible rotators with one internal degree of freedom. This approach integrates a stochastic description of coupled global tumbling and internal torsional motion, quantum chemical calculations of the local potential and the local geometry at the site of the restricted torsion, and hydrodynamics-based calculations of the diffusive properties. The method is applied to the disaccharide beta-D-Glcp-(1-->6)-alpha-D-[6-(13)C]-Manp-OMe dissolved in a DMSO d(6)/D(2)O cryosolvent. The experimental NMR relaxation parameters, associated with the (13)CH(2) probe residing at the glycosidic linkage, include (13)C T(1) and T(2) and (13)C-{(1)H} nuclear Overhauser enhancement (NOE) as well as longitudinal and transverse dipole-dipole cross-correlated relaxation rates, acquired in the temperature range of 253-293 K. These data are predicted successfully by the new theory with only the H-C-H angle allowed to vary. Previous attempts to fit these data using mode-decoupling models failed. PMID- 20025330 TI - Solvation and microscopic properties of ionic liquid/acetonitrile mixtures probed by high-pressure infrared spectroscopy. AB - The microscopic features of binary mixtures formed by an ionic liquid (EMI(+)TFSA(-) or EMI(+)FSA(-)) and a molecular liquid (acetonitrile or methanol) have been investigated by high-pressure infrared spectroscopy. On the basis of its responses to changes in pressure and concentration, the imidazolium C-H appears to exist at least in two different forms, i.e., isolated and associated structures. The weak band at approximately 3102 cm(-1) should be assigned to the isolated structure. CD(3)CN can be added to change the structural organization of ionic liquids. The compression of an EMI(+)TFSA(-)/CD(3)CN mixture leads to the increase in the isolated C-H band intensity. Nevertheless, the loss in intensity of the isolated structures was observed for EMI(+)FSA(-)/CD(3)CN mixtures as the pressure was elevated. In other words, the associated configuration is favored with increasing pressure by debiting the isolated form for EMI(+)FSA(-)/CD(3)CN mixtures. The stronger C-H...F interactions in EMI(+)FSA(-) may be one of the reasons for the remarkable differences in the pressure-dependent results of EMI(+)TFSA(-) and EMI(+)FSA(-). PMID- 20025331 TI - Pressure-energy correlations in liquids. III. Statistical mechanics and thermodynamics of liquids with hidden scale invariance. AB - In this third paper of the series, which started with Bailey et al. [J. Chem. Phys. 129, 184507 (2008); ibid. 129, 184508 (2008)], we continue the development of the theoretical understanding of strongly correlating liquids--those whose instantaneous potential energy and virial are more than 90% correlated in their thermal equilibrium fluctuations at constant volume. The existence of such liquids was detailed in previous work, which identified them, based on computer simulations, as a large class of liquids, including van der Waals liquids but not, e.g., hydrogen-bonded liquids. We here discuss the following: (1) the scaling properties of inverse power-law and extended inverse power-law potentials (the latter includes a linear term that "hides" the approximate scale invariance); (2) results from computer simulations of molecular models concerning out-of-equilibrium conditions; (3) ensemble dependence of the virial/potential energy correlation coefficient; (4) connection to the Gruneisen parameter; and (5) interpretation of strong correlations in terms of the energy-bond formalism. PMID- 20025332 TI - Pressure-energy correlations in liquids. IV. "Isomorphs" in liquid phase diagrams. AB - This paper is the fourth in a series devoted to identifying and explaining the properties of strongly correlating liquids, i.e., liquids where virial and potential energy correlate better than 90% in their thermal equilibrium fluctuations in the NVT ensemble. For such liquids we here introduce the concept of "isomorphic" curves in the phase diagram. A number of thermodynamic, static, and dynamic isomorph invariants are identified. These include the excess entropy, the isochoric specific heat, reduced-unit static and dynamic correlation functions, as well as reduced-unit transport coefficients. The dynamic invariants apply for both Newtonian and Brownian dynamics. It is shown that after a jump between isomorphic state points the system is instantaneously in thermal equilibrium; consequences of this for generic aging experiments are discussed. Selected isomorph predictions are validated by computer simulations of the Kob Andersen binary Lennard-Jones mixture, which is a strongly correlating liquid. The final section of the paper relates the isomorph concept to phenomenological melting rules, Rosenfeld's excess entropy scaling, Young and Andersen's approximate scaling principle, and the two-order parameter maps of Debenedetti and co-workers. This section also shows how the existence of isomorphs implies an "isomorph filter" for theories for the non-Arrhenius temperature dependence of viscous liquids' relaxation time, and it explains isochronal superposition for strongly correlating viscous liquids. PMID- 20025333 TI - Ultrafast conversions between hydrogen bonded structures in liquid water observed by femtosecond x-ray spectroscopy. AB - We present the first femtosecond soft x-ray spectroscopy in liquids, enabling the observation of changes in hydrogen bond structures in water via core-hole excitation. The oxygen K-edge of vibrationally excited water is probed with femtosecond soft x-ray pulses, exploiting the relation between different water structures and distinct x-ray spectral features. After excitation of the intramolecular OH stretching vibration, characteristic x-ray absorption changes monitor the conversion of strongly hydrogen-bonded water structures to more disordered structures with weaker hydrogen-bonding described by a single subpicosecond time constant. The latter describes the thermalization time of vibrational excitations and defines the characteristic maximum rate with which nonequilibrium populations of more strongly hydrogen-bonded water structures convert to less-bonded ones. On short time scales, the relaxation of vibrational excitations leads to a transient high-pressure state and a transient absorption spectrum different from that of statically heated water. PMID- 20025334 TI - Shimmed matching pulses: simultaneous control of rf and static gradients for inhomogeneity correction. AB - Portable NMR systems generally suffer from poor field homogeneity and are therefore used more commonly for imaging and relaxation measurements rather than for spectroscopy. In recent years, various approaches have been proposed to increase the sample volume that is usable for spectroscopy. These include approaches based on manual shimming and those based on clever combinations of modulated radio frequency and gradient fields. However, this volume remains small and, therefore, of limited utility. We present improved pulses designed to correct for inhomogeneous dispersion across wide ranges of frequency offsets without eliminating chemical shift or spatial encoding. This method, based on the adiabatic double passage, combines the relatively larger corrections available from spatially matched rf gradients [C. Meriles et al., J. Magn. Reson. 164, 177 (2003)]. with the adjustable corrections available from time-modulated static field gradients [D. Topgaard et al., Proc. Natl. Acad. Sci. U.S.A. 101, 17576 (2004)]. We explain the origins of these corrections with a theoretical model that simplifies and expedites the design of the pulse waveforms. We also present a generalized method for evaluating and comparing pulses designed for inhomogeneity correction. Experiments validate this method and support simulations that offer new possibilities for significantly enhanced performance in portable environments. PMID- 20025335 TI - Molecular polarizabilities in aqueous proton transfer reactions. AB - Dipole polarizabilities of individual ions and molecules are computed from first principles in three condensed-phase systems: pure water, pure hydrofluoric acid, and an equimolar mixture of water and hydrofluoric acid in which HF is mostly ionized. We find that the polarizability of fluorine and oxygen centers varies linearly with the value of the bond order, which measures the local degree of advancement of the ionization reaction F-H+H(2)O<==>[F(delta ).H.(delta+)OH(2)]<==>F(-)+H(3)O(+). This observation explains the validity of the Lorentz-Lorenz formula for mixtures of acids and water and could have important practical consequences concerning the construction of empirical polarizable reactive force fields. Our results are consistent with the Mulliken charge-transfer picture of proton transfer reactions. The present results also suggest that the average isotropic polarizability of a chemical entity changes substantially only when that entity is involved in charge-transfer processes. PMID- 20025336 TI - The information content of electron flow through adsorbed molecular monolayers. AB - We point to the ability of noncontact measurements of electron transport via self assembled monolayers to provide chemical, A-resolved information about the underlying molecule. A conceptual framework is presented to model a current flow of soft electrons through a molecular monolayer to a substrate and explore the information content of this and other emerging noncontact measurements. A numerical scheme is developed where advantage is taken of the split-operator formalism to propagate the incident electronic wave function over a suitable periodic potential energy surface representing the self-assembled monolayer. The (experimentally observable) potential difference introduced by the transmitted electrons is extracted from the time-averaged electron density using the Poisson equation of classical electrostatics. PMID- 20025337 TI - X-ray study of the structure of liquid water. AB - The radial distribution function (RDF) of liquid water has been determined by x ray scattering. Using a free-standing water jet and a diffracted beam analyzer to separate the Compton from the elastic scattering experimentally, several significant sources of error have been eliminated in the experiment. These have enabled the collection of quality data out to 19.7 A(-1). Data were collected both at 278 K and at 296 K. Our RDF presents fine structure, particularly, a distinct peak at 3.4 A and a peak split at 4.5 A. These features cannot be interpreted by a simple tetrahedral configuration suggested by continuum models. The uncertainties due to the truncation effects and the choice of scattering factors are found to have little influence on the RDF at intermolecular distances. PMID- 20025338 TI - From thermodynamic cell models to partitioning cellular automata for diffusion in zeolites. I. Structure of the algorithm. AB - In the study of adsorption of simple adsorbates in microporous materials like zeolites, thermodynamic models of small grand-canonical cells with very local interactions [e.g., see K. G. Ayappa, J. Chem. Phys. 111, 4736 (1999)] have been proven to be able to produce thermodynamic properties in very good agreement with the results of experiments and atomistic simulations. In this paper we present in details the structure and implementation of a thermodynamic partitioning cellular automaton (PCA) devised as a dynamical version of thermodynamic cell models and proposed as an easy environment to perform coarse-grained simulations of adsorption/diffusion of simple interacting molecules in microporous materials. Local evolution rules and memory effects are introduced to make our PCA able to complete the static picture provided by thermodynamic cell models with the simulation of transport properties. PMID- 20025339 TI - From thermodynamic cell models to partitioning cellular automata for diffusion in zeolites. II. Static and dynamic properties. AB - In this second paper we exploit our thermodynamic partitioning cellular automaton (PCA) developed in Paper I [Pazzona et al., J. Chem. Phys. 131, 234703 (2009)] to study interacting molecules adsorbed in microporous materials. We present a mean field theory of the single cell model at equilibrium followed by a detailed description of the procedure we propose to calculate the chemical potential in the canonical ensemble. Finally we use our approach to simulate transport properties starting from the parameterization devised by Ayappa [J. Chem. Phys. 111, 4736 (1999)] to reproduce the adsorption properties of xenon in zeolite NaA. We report how the correlations included in the PCA evolution rule affect the estimated self-diffusion coefficient. PMID- 20025340 TI - Steady-state composition of a two-component gas bubble growing in a liquid solution: self-similar approach. AB - The paper presents an analytical description of the growth of a two-component bubble in a binary liquid-gas solution. We obtain asymptotic self-similar time dependence of the bubble radius and analytical expressions for the nonsteady profiles of dissolved gases around the bubble. We show that the necessary condition for the self-similar regime of bubble growth is the constant, steady state composition of the bubble. The equation for the steady-state composition is obtained. We reveal the dependence of the steady-state composition on the solubility laws of the bubble components. Besides, the universal, independent from the solubility laws, expressions for the steady-state composition are obtained for the case of strong supersaturations, which are typical for the homogeneous nucleation of a bubble. PMID- 20025341 TI - Polarization-induced switching effect in graphene nanoribbon edge-defect junction. AB - With nonequilibrium Green's function approach combined with density functional theory, we perform an ab initio calculation to investigate transport properties of graphene nanoribbon (GNR) junctions self-consistently. Tight-binding approximation is applied to model the zigzag (ZGNR) electrodes, and its validity is confirmed in comparison to the GAUSSIAN03 periodic boundary condition calculation result of the same system. The origin of abnormal jump points usually appearing in the transmission spectrum is explained with the detailed tight binding ZGNR band structure. Transport property of an edge-defect ZGNR junction is investigated, and the tunable tunneling current can be sensitively controlled by transverse electric fields. PMID- 20025342 TI - Percolation in polymer-solvent systems: a Monte Carlo study. AB - In this study we investigated the percolation in the system containing long flexible polymer chains. The system also contained explicit solvent molecules. The polymer chains were represented by linear sequences of lattice points restricted to a two-dimensional triangular lattice. The Monte Carlo simulations were performed applying the cooperative motion algorithm. The percolation thresholds and the critical exponents of chains and solvent molecules were determined. The influence of the chain length on the percolation was discussed. It was shown that the percolation threshold decreased strongly with the chain length, which is closely connected to changes in chains' structure with the decreasing polymer concentration. The critical exponent beta for all chains under consideration and for solvent molecules was found almost constant and close to the theoretical value 5/36. PMID- 20025343 TI - Simulation study on the formation of vesicle and influence of solvent. AB - The spontaneous vesicle formation of amphiphiles A(1)B(3)s in solution has been studied using a lattice dynamic Monte Carlo simulation. The amphiphilic property of A(1)B(3) chain is represented by the attraction between B-B beads. Two pathways of vesicle formation are observed for different strengths of repulsive interaction between B bead and solvent. In the first pathway, a bilayer disk is aggregated in a randomly dispersed system. It then bends and encapsulates solvents, and finally closes up to form a vesicle. In the second pathway, spherical aggregates are quickly assembled and grow through the coalescence of aggregates or the evaporation-condensation-like process. Then a vesicle is formed when A beads and solvents enter into the center of the aggregate. PMID- 20025344 TI - On the anisotropy of lattice polymers. AB - We quantified the anisotropy of various lattice models used in polymer simulations based on two quantities: the Fourier transform of the normalized Boltzmann factor of allowable bonds on a lattice (which is the central quantity for describing lattice chain conformations) and the bulk lamellar period at the mean-field order-disorder transition (ODT) of symmetric diblock copolymers on a lattice (which is pertinent to the study of microphase separation). This allowed us to compare the anisotropy of different lattices and to design new lattice models that minimize the quantified anisotropy. A lattice field theory based on the same Hamiltonian used in the newly proposed fast lattice Monte Carlo simulations [Q. Wang, Soft Matter 5, 4564 (2009)] is also presented, along with some mean-field results on the ODT of symmetric diblock copolymers on these lattices obtained under the random-phase approximation. PMID- 20025345 TI - Water permeability of poly(ethylene terephthalate): a grand canonical ensemble molecular dynamics simulation study. AB - In this work, our previous simulation method on the calculation of solubility of nonpolar solutes in nonpolar polymers [H. Eslami and F. Muller-Plathe, Macromolecules 40, 6413 (2007)] has been extended to the case of solubility calculation for water, as a polar penetrant, in poly(ethylene terephthalate), as a polar polymer. The chemical potentials of water in the polymer phase and in the gas phase have been calculated by employing our grand canonical ensemble molecular dynamics simulation method [H. Eslami and F. Muller-Plathe, J. Comput. Chem. 28, 1763 (2007)]. In this paper it is shown that performing just two independent simulations, one in the polymer phase and one in the vapor phase, in the grand canonical ensemble, is sufficient to calculate the phase coexistence point. The calculated solubilities, diffusion coefficients, and permeability coefficients are in good agreement with experimental data. Also the calculated glass transition temperature of the wet polymer is shown to be in a very good agreement with experiment. PMID- 20025346 TI - Mesoscale hydrodynamic simulation of short polyelectrolytes in electric fields. AB - The dynamical, conformational, and transport properties of short flexible polyelectrolytes are studied in salt-free solution under the influence of an external electric field taking hydrodynamic interactions into account. A coarse grained polymer model is applied and the multiparticle collision dynamics approach is adopted to account for the solvent. We consider various Coulomb interaction strengths and polymer lengths. The scaling behavior of the diffusion coefficient with respect to polymer length strongly depends on the charge interaction strength due to changes in the polyelectrolyte conformations by counterion condensation. The diffusion coefficients at weak Coulomb interaction strengths can well be described by the expression of rodlike objects, whereas at stronger electrostatic interactions, we observe a Zimm-like behavior. The polyelectrolyte mobilities obey the Nernst-Einstein relation at weak charge interactions. For large interaction strengths and long polymers, the mobilities are independent of molecular weight because of the screening of the intramolecular Coulomb interactions by counterions. Our studies demonstrated that the dynamics of polyelectrolytes in dilute solution and in an external field is governed by an interplay of hydrodynamic interactions and counterion condensation. PMID- 20025347 TI - Temperature influence on the crystallization of polyethylene/fullerene nanocomposites: molecular dynamics simulation. AB - The crystallization of polyethylene/fullerene (PE/C60) nanocomposites with different fullerene content was investigated at different temperatures by means of molecular dynamics simulation. It is found that there is a critical temperature for PE/C60 nanocomposite crystallization. The high C60 content makes the low critical temperature. Crystallinity of the equilibrium conformations of PE/5C60 gradually decreases with increasing temperature. Distributions of the dihedral angle along the PE chain, the radius of gyration, and its three Cartesian components are used to characterize changes in the shape and structure of PE chain as temperature increases. PMID- 20025348 TI - Potential theory of the depletion interaction in the colloid-polymer mixtures. AB - By developing and making use of the potential theory of the polymer-mediated interaction between spherical colloids, we investigate the many-body effects on the depletion interaction among these colloids in the colloid-polymer mixture. As our main results, we obtain analytic expressions for the polymer end density in the presence of the colloids and the many-body depletion potential acting between these colloids in the "protein" limit. We present a comparison of our theoretical findings with the results of recent computer simulations. PMID- 20025349 TI - Thermal and mechanical multistate folding of ribonuclease H. AB - Two different classes of experimental techniques exist by which protein folding mechanisms are ascertained. The first class, of which circular dichroism is an example, probes thermally-induced folding. The second class, which includes atomic force microscopy and optical tweezers, measures mechanically-induced folding. In this article, we investigate if proteins fold/unfold via the same mechanisms both thermally and mechanically. We do so using Ribonuclease H, a protein that has been shown to fold through a three-state mechanism using both types of experimental techniques. A detailed, molecular-level description of the states involved in thermal and mechanical folding shows that mechanisms for both types are globally similar, but small difference exist in the most unfolded conformations. Comparison to previous work suggests a universal folding behavior for proteins with a core helical bundle. PMID- 20025350 TI - The effective diffusion coefficient of a small molecule in a two-phase gel medium. AB - Using simple theoretical arguments and exact numerical lattice calculations, Hickey et al. [J. Chem. Phys. 124, 204903 (2006)] derived and tested an expression for the effective diffusion coefficient of a probe molecule in a two phase medium consisting of a hydrogel with large gel-free inclusions. Although providing accurate predictions, this expression neglects important characteristics that such two-phase systems can present. In this article, we extend the previously derived expression in order to include local interactions between the gel and the analyte, interfacial effects between the main phase and the inclusions, and finally a possible incomplete separation between the two phases. We test our new, generalized expressions using exact numerical calculations. These generalized equations should be a useful tool for the development of novel multiphase systems for specific applications, such as drug delivery platforms. PMID- 20025351 TI - Specificity and completion time distributions of biochemical processes. AB - In order to produce specific complex structures from a large set of similar biochemical building blocks, many biochemical systems require high sensitivity to small molecular differences. The first and most common model used to explain this high specificity is kinetic proofreading, which has been extended to a variety of systems from detection of DNA mismatch to cell signaling processes. While the specification properties of kinetic proofreading models are well known and were studied in various contexts, very little is known about their temporal behavior. In this work, we study the dynamical properties of discrete stochastic two-branch kinetic proofreading schemes. Using the Laplace transform of the corresponding chemical master equation, we obtain an analytical solution for the completion time distribution. In particular we provide expressions for the specificity as well as the mean and variance of the process completion times. We also show that, for a wide range of parameters, a process distinguishing between two different products can be reduced to a much simpler three-point process. Our results allow for the systematic study of the interplay between specificity and completion times, as well as testing the validity of the kinetic proofreading model in biological systems. PMID- 20025352 TI - Internal friction and mode relaxation in a simple chain model. AB - We consider the equilibrium relaxation properties of the end-to-end distance and of the principal components in a one-dimensional polymer chain model with nonlinear interaction between the beads. While for the single-well potentials these properties are similar to the ones of a Rouse chain, for the double-well interaction potentials, modeling internal friction, they differ vastly from the ones of the harmonic chain at intermediate times and intermediate temperatures. This minimal description within a one-dimensional model mimics the relaxation properties found in much more complex polymer systems. Thus, the relaxation time of the end-to-end distance may grow by orders of magnitude at intermediate temperatures. The principal components (whose directions are shown to coincide with the normal modes of the harmonic chain, whatever interaction potential is assumed) not only display larger relaxation times but also subdiffusive scaling. PMID- 20025353 TI - The form factor of H-comb polymers. AB - A Monte Carlo pivot algorithm is employed to investigate the form factor of continuum, tangent hard sphere H-comb polymers in both the ideal and excluded volume regimes. The simulated form factors for 241 and 931 "bead" ideal H-combs are essentially the same. The results for these polymers are in excellent agreement with the theoretical prediction. There is only a slight difference in the form factor between the ideal and excluded volume regimes at larger values of distance. PMID- 20025355 TI - National cost efficiency of supported employees with intellectual disabilities: 2002 to 2007. AB - The cost efficiency of supported employees with intellectual disabilities who were served by vocational rehabilitation agencies throughout the United State from 2002 to 2007 was explored. Findings indicate that, on average, supported employees with intellectual disabilities were cost-efficient from the taxpayers' perspective regardless of whether they had secondary disabilities. In addition, no changes in cost efficiency were found during the period investigated. The data, however, did demonstrate considerable variability in cost efficiency throughout the United States and its territories. PMID- 20025356 TI - Mental health problems in adults with Williams syndrome. AB - Although many researchers have investigated emotional and behavioral difficulties in individuals with Williams syndrome, few have used standardized diagnostic assessments. We examined mental health problems in 92 adults with Williams syndrome using the Psychiatric Assessment Schedule for Adults with Developmental Disabilities-PAS-ADD (Moss, Goldberg, et al., 1996). Factors potentially associated with mental health problems were also explored. The PAS-ADD identified mental health problems in 24% of the sample. The most common were anxiety (16.5%) and specific phobias (12%). Other diagnoses included depression, agoraphobia, and social phobia. No association was found between the presence of mental health problems and either individual (e.g., age, IQ, language level) or external (life events) variables. PMID- 20025357 TI - Socioeconomic circumstances and risk of psychiatric disorders among parents of children with early cognitive delay. AB - Results of previous research suggest that parents of children with intellectual disabilities are at increased risk of psychological distress and psychiatric disorder. Secondary analysis of the Millennium Cohort Study in the United Kingdom indicated that controlling for between-group differences in socioeconomic circumstances reduced the differences in probable psychiatric disorder to non significance for fathers and markedly attenuated the strength of the relationship for mothers, especially for those with children who have less severe early cognitive delay. These data are consistent with the notion that greater than expected risk for psychiatric disorder among parents of young children with early cognitive delay is related to aspects of the socioeconomic circumstances of families. PMID- 20025358 TI - Perceptions of body image by persons with Prader-Willi syndrome and their parents. AB - Prader-Willi syndrome is a genetic disorder characterized by obesity. The Figure Rating Scale (Stunkard, Sorensen, & Schulsinger, 1983) was completed by 43 individuals with this syndrome to determine their level of dissatisfaction with their body. Their parents also completed this scale regarding their child to determine whether they were dissatisfied with their child's body status. Results showed that individuals with Prader-Willi syndrome were dissatisfied with their body. Parents also were dissatisfied with their child's body. Results of this study demonstrate that the responses of persons with Prader-Willi syndrome on the Figure Rating Scale show significant discrepancies between how they think they look and how they wished they looked. PMID- 20025359 TI - Maternal responsivity predicts language development in young children with fragile X syndrome. AB - The relationship between early maternal responsivity and later child communication outcomes in young children with fragile X syndrome was investigated. Data were obtained from 55 mother-child dyads over a 36-month period. Performance data were obtained at each measurement point from video observations of four different contexts. These were coded for (a) child communication behaviors, (b) parent responsivity, and (c) behavior management behaviors. Results indicate that early maternal responsivity predicts the level of four important child language outcomes at 36 months of age after controlling for child developmental level and autism symptomology. PMID- 20025361 TI - Alcohol-induced disinhibition expectancies and impaired control as prospective predictors of problem drinking in undergraduates. AB - Trait disinhibition is associated with problem drinking and alcohol drinking can bring about a state of disinhibition. It is unclear however, if expectancies of alcohol-induced disinhibition are unique predictors of problem drinking. Impaired control (i.e., difficulty in limiting alcohol consumption) may be related to disinhibition expectancies in that both involve issues of control related to alcohol use. Data from a prospective survey of undergraduates assessed during freshman (N = 337) and senior year (N = 201) were analyzed to determine whether subscales of the Drinking-Induced Disinhibition Scale (Leeman, Toll, & Volpicelli, 2007) and the Impaired Control Scale (Heather et al., 1993) predicted unique variance in heavy episodic drinking and alcohol-related problems. In Time 1 cross-sectional models, Dysphoric disinhibition expectancies predicted alcohol related problems and impaired control predicted both alcohol-related problems and heavy episodic drinking. In prospective models, Time 1 impaired control predicted Time 2 alcohol-related problems and Time 1 Euphoric/social Disinhibition expectancies predicted Time 2 heavy episodic drinking. These findings suggest that expectancies of alcohol-induced disinhibition and impaired control predict unique variance in problem drinking cross-sectionally and prospectively, and that these phenomena should be targeted in early intervention efforts. PMID- 20025363 TI - Academic and social motives and drinking behavior. AB - This longitudinal study of 1,447 first-time college students tested separate time varying covariate models of the relations between academic and social motives/behaviors and alcohol use and related problems from senior year of high school through the end of the second year in college. Structural equation models identified small but significant inverse relations between academic motives/behaviors and alcohol use across all time points, with relations of somewhat larger magnitude between academic motives/behaviors and alcohol-related problems across all semesters other than senior year in high school. At all time points, there were much larger positive relations between social motives/behaviors and alcohol use across all semesters, with smaller but significant relations between social motives/behaviors and alcohol-related problems. Multi-group models found considerable consistency in the relations between motives/behaviors and alcohol-related outcomes across gender, race/ethnicity, and family history of alcohol problems, although academic motives/behaviors played a stronger protective role for women, and social motives were a more robust risk factor for Caucasian and Latino students and individuals with a positive family history of alcohol problems. Implications for alcohol prevention efforts among college students are discussed. PMID- 20025364 TI - Drink less or drink slower: the effects of instruction on alcohol consumption and drinking control strategy use. AB - Brief alcohol interventions often involve recommendations to use drinking control strategies. However, little is known about the functional effect of these strategies on alcohol use. This prospective study employed an experimental design to evaluate the relationship between strategy use and alcohol consumption. The differential effects of instructions to increase the use of strategies or to reduce alcohol consumption were compared to self-monitoring (SM) only. Undergraduate drinkers were randomized into 3 conditions: SM plus strategy increase (SI; n = 61), SM plus alcohol reduction (AR; n = 60), and SM control (SM; n = 56). Participants in the AR group reduced their alcohol use over 2 weeks, while those in the SI group did not drink less. Participants in the SI group increased strategy use over time, whereas the AR group increased use of some strategies but not others. These results indicate that increasing use of drinking control strategies does not necessarily result in reduced drinking. Furthermore, all strategies are not equal in their association with alcohol consumption; if the goal is alcohol reduction, type of strategy recommended may be important. PMID- 20025365 TI - Refining measurement in the study of social anxiety and student drinking: who you are and why you drink determines your outcomes. AB - This study investigated inconsistencies in the literature regarding social anxiety and problematic drinking among college students. One hundred eighteen students (61% women) who experience anxiety in social or performance situations completed measures of social anxiety and a modified Timeline Followback that assessed the psychological context of drinking episodes and alcohol-related consequences. Results suggest that men who experience severe social anxiety drink less alcohol than men with lower levels of anxiety, whereas women high in social anxiety are likely to experience more alcohol-related consequences per drinking episode than women low in social anxiety, despite drinking similar amounts of alcohol. In addition, women with high social anxiety were found to experience more alcohol-related consequences than men with high social anxiety. These findings suggest that the inconsistencies noted in the literature on drinking to cope with social anxiety and alcohol-related consequences may reflect methodological differences and the failure to consider gender. PMID- 20025366 TI - What makes group MET work? A randomized controlled trial of college student drinkers in mandated alcohol diversion. AB - Nationally, college drinkers exhibit the highest rates of alcohol consumption and represent the largest percentage of problem drinkers. Group motivational enhancement therapy (GMET) has been found to catalyze problem drinking reductions among college student samples. Although research supporting the use of single session GMET in college samples (general and mandated) is emergent, no studies have evaluated a comprehensive model of the potential active ingredients of this group intervention. College students (N = 206; 88% White; 63% men; M age = 18.6) mandated to a university alcohol diversion program were randomly assigned to 1 of 3 conditions: the standard-of-care 2-session "Focus on Alcohol Concerns" education group (FAC), a single GMET, or a single alcohol information-only control group (AI) to evaluate the role of 5 putative mediators: readiness to change, self-efficacy, perceived risk, norm estimates, and positive drinking expectancies. At 3- and 6-month follow-ups, GMET students demonstrated greater reductions in problem drinking outcomes (drinks per drinking day, hazardous drinking symptoms, and alcohol-related problems). Of the 5 mediators proposed, only self-efficacy emerged as a significant mediator. PMID- 20025367 TI - A stress-coping profile of opioid dependent individuals entering naltrexone treatment: a comparison with healthy controls. AB - BACKGROUND: Stress is known to increase addiction vulnerability and risk of relapse to substance use. PURPOSE & METHOD: We compared opioid dependent individuals entering naltrexone treatment (n = 57) with healthy controls (n = 75) on measures of stress, coping, and social support and examined the relative contribution of group membership, coping, and social support to stress within the sample. Analyses of variance (ANOVA) and covariance (ANCOVA), and stepwise multiple regression were conducted. RESULTS: Compared with controls, opioid dependent subjects reported greater stress, less use of adaptive coping, but comparable use of maladaptive/avoidant coping. No group differences were found with respect to social support. Perceived stress was predicted by group membership, low social support, and greater use of maladaptive/avoidant coping, and the prediction by social support and maladaptive/avoidant coping did not differ by group. CONCLUSION: Opioid dependent individuals entering naltrexone treatment experience higher levels of stress and report less use of adaptive coping strategies when compared with controls. Group membership, maladaptive/avoidant coping, and social support independently contribute to perceived stress. Findings suggest that novel treatment approaches that decrease maladaptive/avoidant coping and improve social support are important aspects of decreasing stress during early recovery from opioid addiction. PMID- 20025369 TI - Similarity in cigarette smoking attracts: a prospective study of romantic partner selection by own smoking and smoker prototypes. AB - In the current research, we used a multiwave longitudinal design to examine how young adults' own smoking and smoker prototypes are associated with selection of romantic partners over time. Results indicate that participants who smoke, versus participants who do not smoke, and participants who have a more positive prototype of the typical smoker are more likely to initiate a romantic relationship with someone who smokes and who has greater perceived approval for smoking. Among participants who smoke, higher levels of smoking are associated with initiating a relationship with a romantic partner who smokes more and approves of smoking more. The findings suggest some important aspects of romantic partner selection effects in terms of what is selected for, partner smoking and approval, and key young adult variables that contribute to selection, such as participant's own smoking and smoker prototype. PMID- 20025368 TI - Tobacco, marijuana, and sensation seeking: comparisons across gay, lesbian, bisexual, and heterosexual groups. AB - This study examined patterns of smoked substances (cigarettes and marijuana) among heterosexuals, gays, lesbians, and bisexuals based on data from the 2000 National Alcohol Survey, a population-based telephone survey of adults in the United States. We also examined the effect of bar patronage and sensation seeking/impulsivity (SSImp) on tobacco and marijuana use. Sexual orientation was defined as lesbian or gay self-identified, bisexual self-identified, heterosexual self-identified with same-sex partners in the past 5 years, and exclusively heterosexual (heterosexual self-identified, reporting no same-sex partners). Findings indicate that bisexual women and heterosexual women reporting same-sex partners had higher rates of cigarette smoking than exclusively heterosexual women. Bisexual women, lesbians, and heterosexual women with same-sex partners also used marijuana at significantly higher rates than exclusively heterosexual women. Marijuana use was significantly greater and tobacco use was elevated among gay men compared with heterosexual men. SSImp was associated with greater use of both of these substances across nearly all groups. Bar patronage and SSImp did not buffer the relationship between sexual identity and smoking either cigarettes or marijuana. These findings suggest that marijuana and tobacco use differ by sexual identity, particularly among women, and underscore the importance of developing prevention and treatment services that are appropriate for sexual minorities. PMID- 20025370 TI - Effects of transdermal nicotine on symptoms, moods, and cardiovascular activity in the everyday lives of smokers and nonsmokers with attention deficit/hyperactivity disorder. AB - The aim of the study was to test the self-medication hypothesis by examining the effects of nicotine in the everyday lives of smokers and nonsmokers with attention-deficit/hyperactivity disorder (ADHD). Fifty-two adults with ADHD (25 abstinent smokers and 27 nonsmokers) participated in a double-blind placebo controlled study with one nicotine patch condition and one placebo patch condition in counterbalanced order. Each condition continued for two consecutive days in which patches were administered each morning. The effects of nicotine on ADHD symptoms, moods, and side effects were assessed with electronic diaries. Cardiovascular activity was recorded with ambulatory blood pressure monitors and physical activity was monitored with actigraphs. Nicotine reduced reports of ADHD symptoms by 8% and negative moods by 9%, independent of smoking status. In addition, nicotine increased cardiovascular activity during the first 3 to 6 hours after nicotine patch administration. The results support the self medication hypothesis for nicotine in adults with ADHD and suggest that smoking cessation and prevention efforts for individuals with ADHD will need to address both the symptom reducing and mood enhancing effects of nicotine. PMID- 20025371 TI - The Autonomy Over Smoking Scale. AB - Our goal was to create an instrument that can be used to study how smokers lose autonomy over smoking and regain it after quitting. The Autonomy Over Smoking Scale was produced through a process involving item generation, focus-group evaluation, testing in adults to winnow items, field testing with adults and adolescents, and head-to-head comparisons with other measures. The final 12-item scale shows excellent reliability (alphas = .91-.97), with a one-factor solution explaining 59% of the variance in adults and 61%-74% of the variance in adolescents. Concurrent validity was supported by associations with age of smoking initiation, lifetime use, smoking frequency, daily cigarette consumption, history of failed cessation, Hooked on Nicotine Checklist scores, and Diagnostic and Statistical Manual of Mental Disorder (4th ed., text rev.; American Psychiatric Association, 2000) nicotine dependence criteria. Potentially useful features of this new instrument include (a) it assesses tobacco withdrawal, cue induced craving, and psychological dependence on cigarettes; (b) it measures symptom intensity; and (c) it asks about current symptoms only, so it could be administered to quitting smokers to track the resolution of symptoms. PMID- 20025372 TI - Surfing the urge: brief mindfulness-based intervention for college student smokers. AB - The current study investigates effects of a brief mindfulness-based instruction set, based on Marlatt's "urge surfing" technique (Marlatt & Gordon, 1985), on smoking-related urges and behavior. Undergraduate smokers (N = 123) who were interested in changing their smoking, but not currently involved in a cessation program, participated in a cue exposure paradigm designed to elicit urges to smoke. They were randomly assigned either to a group receiving brief mindfulness based instructions or to a no-instruction control group. Results suggest that groups did not differ significantly on measures of urges. However, those in the mindfulness group smoked significantly fewer cigarettes over a 7-day follow-up period as compared to those in the control group. These findings suggest that the mindfulness techniques may not initially reduce urges to smoke but may change the response to urges. The study provides preliminary data for future studies examining both mechanisms and effectiveness of mindfulness-based interventions for cigarette smoking. PMID- 20025373 TI - Randomized controlled trial of a cognitive-behavioral motivational intervention in a group versus individual format for substance use disorders. AB - Although group therapy is widely used for individuals with substance use disorders (SUDs), randomized clinical trials (RCTs) comparing the same treatment in a group versus individual format are rare. This paper presents the results of a RCT comparing guided self-change (GSC) treatment, a cognitive-behavioral motivational intervention, conducted in a group versus individual format with 212 alcohol abusers and 52 drug abusers who voluntarily sought outpatient treatment. Treatment outcomes demonstrated significant and large reductions in clients' alcohol and drug use during treatment and at the 12-month follow-up, with no significant differences between the group and individual therapy conditions. A therapist time ratio analysis found that it took 41.4% less therapist time to treat clients using the group versus the individual format. Participants' end-of treatment group cohesion scores characterized the groups as having high engagement, low levels of interpersonal conflict, and low avoidance of group work, all desirable group characteristics. These findings suggest that the GSC treatment model was effectively integrated into a brief group treatment format. Health care cost containment compels further evaluations of the efficacy of group treatments for SUDs. PMID- 20025374 TI - Thinking about drinking: need for cognition and readiness to change moderate the effects of brief alcohol interventions. AB - Research has demonstrated the efficacy of brief motivational interventions (BMI) and alcohol expectancy challenge (AEC) in reducing alcohol use and/or problems among college students. However, little is known about variables that may qualify the effectiveness of these approaches. The present analyses tested the hypothesis that need for cognition (NFC), impulsivity/sensation seeking (IMPSS) and readiness to change (RTC) would moderate the effects of BMI and AEC. Participants (N = 335) were heavy drinking college students enrolled in a randomized 2 x 2 factorial study of BMI and AEC. Latent growth curve analyses indicated significant interactions for BMI x NFC and AEC x RTC on alcohol use but not problems. Simple slopes analyses were used to probe these relationships and revealed that higher levels of NFC at baseline were associated with a stronger BMI effect on drinking outcomes over time. Similarly, higher levels of baseline RTC were associated with stronger AEC effects on alcohol use. Future preventive interventions with this population may profit by considering individual differences and targeting approaches accordingly. PMID- 20025375 TI - Preliminary examination of spring break alcohol use and related consequences. AB - The authors examined the extent to which college student drinkers are at risk for experiencing negative alcohol-related consequences during Spring Break. A sample of first-year college student drinkers (N = 726) participated by completing an online survey assessing typical drinking, as well as Spring Break drinking and related consequences. Findings suggest Spring Break drinking was positively associated with alcohol-related consequences during Spring Break, even after controlling for sex and typical drinking. Furthermore, results indicated that typical drinking moderated the relationship between Spring Break drinking and expected zero-values (i.e., not reporting any Spring Break consequences), such that the association between Spring Break drinking and the likelihood of being a zero-score was less evident for those who are typically lighter drinkers. Identifying and examining temporal and contextually relevant events and associated drinking is critical for understanding and ultimately preventing extreme drinking and associated consequences associated with specific events like Spring Break, which place many students at high risk for experiencing acute harm. PMID- 20025376 TI - Alcohol use by undergraduate students on their 21st birthday: predictors of actual consumption, anticipated consumption, and normative beliefs. AB - Recent research has identified celebration of a 21st birthday as an environmental event during which many college students engage in risky levels of alcohol consumption. The current study examined the relationship between personality and different aspects of alcohol use during 21st birthday celebrations: actual amount consumed for those who had turned 21, anticipated amount consumed for those under the age of 21, and normative beliefs regarding the amount other students consume on their 21st birthdays. Sensation seeking and impulsivity both displayed significant bivariate relationships with all three aspects of 21st birthday drinking. Personality traits did not contribute unique variance to actual 21st birthday drinking after the effects of typical alcohol consumption were accounted for in the models. Impulsivity contributed unique variance to models accounting for anticipated drinking and normative beliefs. Additional research is necessary to better understand the role personality variables play on alcohol consumption during 21st birthday celebrations. PMID- 20025377 TI - Prospective associations among alcohol use-related sexual enhancement expectancies, sex after alcohol use, and casual sex. AB - Higher levels of alcohol use have consistently been related to higher rates of sexual risk taking; however, it is not clear whether this relationship is causal. This study examined the concurrent and predictive associations among alcohol use related sexual enhancement expectancies, drinking alcohol before engaging in sex, and casual sex during the transition into emerging adulthood and whether these associations differed for men and women. Data came from 590 men and women who were interviewed 3 times at 6-month intervals after high school. Growth curve analyses indicated that alcohol-related sexual enhancement expectancies were related to casual sex indirectly through drinking before sex but did not predict change in either of these behaviors. However, increases in drinking before sex predicted increases in casual sex over time. The findings provide some support for prevention programs that focus on alcohol-related sexual expectancies to reduce sexually transmitted illnesses among emerging adults. PMID- 20025378 TI - Underage drinking among young adolescent girls: the role of family processes. AB - Guided by family interaction theory, this study examined the influences of psychological, peer, and familial processes on alcohol use among young adolescent girls and assessed the contributions of familial factors. An ethnically diverse sample of 1,187 pairs of girls (M age = 12.83 years), and their mothers completed surveys online. Questionnaires assessed girls' lifetime and recent alcohol use, as well as girls' demographic, psychological, peer, and family characteristics. Hierarchical logistic regression models showed that although girls' drinking was associated with a number of psychological and peer factors, the contributions of family domain variables to girls' drinking were above and beyond that of psychological and peer factors. The interaction analyses further highlighted that having family rules, high family involvement, and greater family communication may offset risks in psychological and peer domains. Study findings underscore the multifaceted etiology of drinking among young adolescent girls and assert the crucial roles of familial processes. Prevention programs should be integrative, target processes at multiple domains, and include work with parents. PMID- 20025380 TI - Acceptance and commitment therapy for smoking cessation: a preliminary study of its effectiveness in comparison with cognitive behavioral therapy. AB - This controlled preliminary trial determined the feasibility and initial effectiveness of a promising behavioral intervention for smoking: Acceptance and Commitment Therapy (ACT). In a quasi-experimental design, the ACT intervention condition used metaphors and experiential exercises focused on personal values to motivate quitting smoking and enhancing the willingness to experience internal cues to smoke (e.g., urges) and abstinence-related internal distress. The comparison condition was cognitive behavioral therapy (CBT)--the current standard in behavioral intervention for smoking cessation. Each treatment was delivered in seven weekly 90-min sessions in a group format to 81 (43 in ACT; 38 in CBT) adult smokers. Results show that the ACT treatment was as feasible as the CBT treatment. They also demonstrate promising evidence of ACT's effectiveness: 30.2% intent-to-treat biochemically-supported 30-day point prevalence at twelve month follow-up, compared with 13.2% in the CBT condition (odds ratio = 5.13; p = .02). Replication in a well-powered, randomized, controlled trial is now needed. PMID- 20025379 TI - Initiating moderate to heavy alcohol use predicts changes in neuropsychological functioning for adolescent girls and boys. AB - This study prospectively examines the influence of alcohol on neuropsychological functioning in boys and girls characterized prior to initiating drinking (N = 76, ages 12-14). Adolescents who transitioned into heavy (n = 25; 11 girls, 14 boys) or moderate (n = 11; 2 girls, 9 boys) drinking were compared with matched controls who remained nonusers throughout the approximately 3-year follow-up period (N = 40; 16 girls, 24 boys). For girls, more past year drinking days predicted a greater reduction in visuospatial task performance from baseline to follow-up, above and beyond performance on equivalent measures at baseline (R2Delta = 10%, p < .05), particularly on tests of visuospatial memory (R2Delta = 8%, p < .05). For boys, a tendency was seen for more past year hangover symptoms to predict worsened sustained attention (R2Delta = 7%, p < .05). These preliminary longitudinal findings suggest that initiating moderately heavy alcohol use and incurring hangover during adolescence may adversely influence neurocognitive functioning. Neurocognitive deficits linked to heavy drinking during this critical developmental period may lead to direct and indirect changes in neuromaturational course, with effects that would extend into adulthood. PMID- 20025381 TI - Cue-specific reactivity in experienced gamblers. AB - To examine whether gambling cue reactivity is cue-specific, 47 scratch-off lottery players and 47 horse race gamblers were presented with video clips of their preferred and nonpreferred modes of gambling, and two control stimuli including an exciting car race and a mental stressor task while heart rates, excitement, and urge to gamble were being measured. Heart rates for both groups of gamblers were highest to the mental stressor and did not differ in response to the other three cues. Excitement for both groups was highest in response to the action cues (horse race and car chase). Urge to gamble was significantly higher for each group to their preferred mode of gambling. A post hoc exploratory analysis comparing social gamblers (n = 54) and probable pathological gamblers (n = 40) revealed a similar pattern of responses. However, pathological gamblers reported overall significantly higher urges to gamble than social gamblers. As urges have been shown to play a pivotal role in addictive behaviors and relapse, the current findings may have implications for the development of gambling problems and relapse after successful treatment. PMID- 20025382 TI - Self-coded indirect memory associations in a brief school-based intervention for substance use suspensions. AB - This study assessed the concurrent validity of self-generated and self-coded substance use associations for marijuana and alcohol use. Grades seven to twelve students were assessed as part of a brief intervention program in lieu of suspension for substance use infractions in school. During the cognitive assessment, students generated memory associations to probes for high-risk situations and desirable outcomes. Later, the participant rated their responses according to categories including both non-risk and substance use. Three different coding methods were compared: (1) conservative codes using clearly unambiguous responses, (2) liberal scores adding ambiguous, but likely responses, and (3) self-coded. Self-coded scores were higher, had stronger correlations with substance use, and were better predictors of substance use and problems than either conservative or liberal coded scores. These findings suggest that self coding may be used to improve concurrent validity, decrease ambiguities in coding, and reduce the cost of measuring memory associations. The present method promises a cost effective and valid measure of indirect substance use cognitions that can be readily adapted for interventions. PMID- 20025383 TI - Language-based measures of mindfulness: initial validity and clinical utility. AB - This study examined relationships among language use, mindfulness, and substance use treatment outcomes in the context of an efficacy trial of mindfulness-based relapse prevention (MBRP) for adults with alcohol and other drug use (AOD) disorders. An expert panel generated two categories of mindfulness language (ML) describing the mindfulness state and the more encompassing "mindfulness journey," which included words describing challenges of developing a mindfulness practice. MBRP participants (n = 48) completed baseline sociodemographic and AOD measures, and participated in the 8-week MBRP program. AOD data were collected during the 4 month follow-up. A word count program assessed the frequency of ML and other linguistic markers in participants' responses to open-ended questions about their postintervention impressions of mindfulness practice and MBRP. Findings supported concurrent validity of ML categories: ML words appeared more frequently in the MBRP manual compared to the 12-step Big Book. Further, ML categories correlated with other linguistic variables related to the mindfulness construct. Finally, predictive validity was supported: greater use of ML predicted fewer AOD use days during the 4-month follow-up. This study provided initial support for ML as a valid, clinically useful mindfulness measure. If future studies replicate these findings, ML could be used in conjunction with self-report to provide a more complete picture of the mindfulness experience. PMID- 20025384 TI - Deficits in other-race face recognition: no evidence for encoding-based effects. AB - The other-race effect (ORE) in face recognition is typically observed in tasks which require long-term memory. Several studies, however, have found the effect early in face encoding (Lindsay, Jack, & Christian, 1991; Walker & Hewstone, 2006). In 6 experiments, with over 300 participants, we found no evidence that the recognition deficit associated with the ORE reflects deficits in immediate encoding. In Experiment 1, with a study-to-test retention interval of 4 min, participants were better able to recognise White faces, relative to Asian faces. Experiment 1 also validated the use of computer-generated faces in subsequent experiments. In Experiments 2 through 4, performance was virtually identical to Asian and White faces in match-to-sample, immediate recognition. In Experiment 5, decreasing target-foil similarity and disrupting the retention interval with trivia questions elicited a re-emergence of the ORE. Experiments 6A and 6B replicated this effect, and showed that memory for Asian faces was particularly susceptible to distraction; White faces were recognised equally well, regardless of trivia questions during the retention interval. The recognition deficit in the ORE apparently emerges from retention or retrieval deficits, not differences in immediate perceptual processing. PMID- 20025385 TI - Featuring familiarity: how a familiar feature instantiation influences categorization. AB - We demonstrate that a familiar looking feature can influence categorization through 2 different routes, depending on whether a person is reliant on abstract feature representations or on concrete feature representations. In 2 experiments, trained participants categorized new category members in a 3-step procedure: Participants made an initial categorization, described the rule-consistent features indicated by the experimenter, and then recategorized the item. Critical was what happened on the second categorization after participants initially categorized an item based on a familiar, but misleading, feature. Participants who were reliant on abstract features most commonly reversed themselves after the rule-consistent features were pointed out, suggesting that the familiar feature had biased attention. Participants who were reliant on concrete feature representations, however, most commonly persisted with the initial response as if the familiar feature were more important than its rivals-the familiar feature biased decision making. PMID- 20025386 TI - [Difficulty of young children of understanding emotion dissimulation]. AB - The authors investigated the understanding of emotion dissimulation in school-age children. Sixty participants were read short stories in which a main character expressed an emotion or hid an emotion from other characters. The participants were asked to identify the emotion felt by the main characters and to indicate the facial expressions they would display. Then they were asked what emotions the main characters felt while they were displaying these expressions, and what the beliefs of the other story characters would be as to the emotion felt by the main characters. The results revealed that children from 5 to 6 years of age have a partial understanding of emotion dissimulation. They were accurate in finding the emotion felt by the main characters when questioned the first time. They were also accurate in choosing the expressions the main characters would display to hide their emotions. However, they were often inaccurate as to the felt emotions of the main characters when questioned the second time. Compared with 9- and 10 year-olds, the younger children had more difficulty understanding the simultaneous character of felt and displayed emotions. Five- and 6-year-olds were also less accurate than the older children when asked to indicate the beliefs of the other characters in stories where felt emotions were hidden. PMID- 20025387 TI - Further evidence that congenitally blind participants react faster to auditory and tactile spatial targets. AB - Congenital blindness is one of the rare human models to explore the role of experience-driven cross-modal compensation after early sensory deprivation. We re examined spatial attention abilities in congenitally blind participants and sighted controls using a paradigm comparable to the one of our previous study (Collignon, Renier, Bruyer, Tranduy, & Veraart, 2006), except that this time the auditory and tactile stimuli were now presented in sequence. Although both groups performed the task with similar accuracy, we observed that blind participants had shorter reaction times than sighted controls for the detection of spatial targets in both sensory modalities. Moreover, this finding held true for both the selective and divided attention conditions. These results not only confirm previous reports on the superiority of the blind during auditory and tactile attention tasks, but also broaden our knowledge of the mechanisms underlying cross-modal compensation. PMID- 20025388 TI - Illusions of competence for phonetically, orthographically, and semantically similar word pairs. AB - Illusions of competence are thought to arise when judgements of learning (JOLs) made in the presence of intact cue-target pairs during study create a "foresight bias," such that JOLs are inflated by the apparent association between a cue and a target, despite the lack of benefit this association has for recall performance. For example, Castel, McCabe, and Roediger (2007) recently demonstrated an illusion of competence for identical word pairs (mouse-mouse). In two experiments, the authors examined possible sources for this over confidence, including phonetic, semantic, and orthographic similarity. An illusion of competence was found for homophones, synonyms, orthographically similar, and unrelated items, whereas no illusion of competence was found for word pairs with a relatively high forward-semantic association. Self-paced study times indicated that encoding fluency was not closely associated with the magnitude of over confidence. Error data revealed participants may have been engaging in strategic responding in order to maximise correct recall. Our results underscore the importance of considering factors that influence both JOLs and recall performance when considering sources of (mis)calibration in absolute accuracy. PMID- 20025389 TI - [Masked orthographic priming in the recognition of written words: empirical data and theoretical prospects]. AB - The present paper reviews the main studies that have been conducted on the effects of masked orthographic priming in written word recognition. Empirical data accumulated over the last two decades are exposed by considering three factors that play a role in the effects of orthographic priming: prime lexicality, prime duration, and target and/or prime orthographic neighbourhood. The theoretical implications of these data are discussed in light of the two major frameworks of visual word recognition, the serial search and the interactive activation. As a whole, the interactive activation hypothesis seems to be more appropriate to account for the empirical data. PMID- 20025390 TI - Flicker is a primitive visual attribute in visual search. AB - At the earliest processing stages, visual stimuli are decomposed by a set of filters tuned to specific values of such attributes as colour, orientation, and motion. These filters have been characterised both neurophysiologically and behaviourally. The single exception is the attribute of flicker that has been characterised neurophysiologically but not behaviourally. Using a visual search paradigm, the authors provide the first behavioural demonstration that flicker is indeed a primitive attribute used by the visual system in stimulus encoding. Consistent with the temporal contrast-sensitivity function, sensitivity to flicker was highest at about 10 Hz and decreased as the flicker rate was either increased or decreased. PMID- 20025391 TI - Increasing the difficulty of response selection does not increase the switch cost. AB - Several theories of task switching assume that basic task processes such as stimulus identification and response selection do not contribute to task-switch costs. This conclusion is mainly based on the finding that stimulus identification manipulations have no influence on the size of the switch cost. The present study tested the influence of response-selection manipulations on the size of the switch cost. The authors manipulated the difficulty of response selection by using a semantically based response-side effect that is associated with numerical-judgement tasks, namely the Spatial Numerical Association of Response Codes (SNARC) effect. The authors observed a SNARC effect and a switch cost, but no interaction between the two: the task-switch cost did not differ between SNARC-compatible and -incompatible responses. The authors conclude that response selection does not contribute to the switch cost on the current trial, which provides further support for the idea that basic task processes and task switch processes are separate. PMID- 20025392 TI - The optic tectum of birds: mapping our way to understanding visual processing. AB - Over the past few decades there has been a massive amount of research on the geniculo-striate visual system in primates. However, studies of the avian visual system have provided a rich source of data contributing to our understanding of visual processing. In this paper we review the connectivity and function of the optic tectum (homolog of the superior colliculus) in birds. We highlight the retinotopic projections that the optic tectum has with the isthmal nuclei, and the functional topographic projections that the optic tectum has with the nucleus rotundus and entopallium (homologs of the pulvinar and extrastriate cortex, respectively) where retinotopy has been sacrificed. This work has been critical in our understanding of basic visual processes including attention, parallel processing, and the binding problem. PMID- 20025394 TI - Introduction to the special section on intraindividual variability and aging. AB - This preface introduces articles that appear in the special section on intraindividual variability and aging and illustrate what intraindividual variability might contribute to the study of development. These articles exemplify the variety of conceptual perspectives, analytical methods, and types of data that are being used to study intraindividual variability and illustrate what the study of intraindividual variability might contribute to the study of development. PMID- 20025395 TI - Time-structured and net intraindividual variability: tools for examining the development of dynamic characteristics and processes. AB - The study of intraindividual variability is the study of fluctuations, oscillations, adaptations, and "noise" in behavioral outcomes that manifest on microtime scales. This article provides a descriptive frame for the combined study of intraindividual variability and aging/development. At the conceptual level, we show that the study of intraindividual variability provides access to dynamic characteristics-construct-level descriptions of individuals' capacities for change (e.g., lability)--and to dynamic processes--the systematic changes that individuals exhibit in response to endogenous and exogenous influences (e.g., regulation). At the methodological level, we review how quantifications of net intraindividual variability and models of time-structured intraindividual variability are used to measure and describe dynamic characteristics and processes. At the research design level, we point to the benefits of measurement burst study designs, wherein data are obtained across multiple time scales, for the study of development. PMID- 20025396 TI - Neural underpinnings of within-person variability in cognitive functioning. AB - Increased intraindividual variability (IIV), reflecting within-person fluctuations in behavioral performance, is commonly observed in aging as well as in select disorders including traumatic brain injury, schizophrenia, attention deficit hyperactivity disorder (ADHD), and dementia. Much recent progress has been made toward understanding the functional significance of IIV in cognitive performance (MacDonald, Nyberg, & Backman, 2006) and biological information processing (Stein, Gossen, & Jones 2005), with parallel efforts devoted to investigating the links between older adults' deficient neuromodulation and their more variable neuronal and cognitive functions (Backman, Nyberg, Lindenberger, Li, & Farde, 2006). Despite these advances in the study of IIV, there has been little empirical examination of underlying neural correlates and virtually no synthesis of extant findings. The present review summarizes the accumulating empirical evidence linking age-related increases in IIV in cognitive performance to neural correlates at anatomical, functional, neuromodulatory, and genetic levels. Computational theories of neural dynamics (e.g., Li, Lindenberger, & Sikstrom, 2001) are also introduced to illustrate how age-related neuromodulatory deficiencies may contribute to increased neuronal noise and render information processing in aging neurocognitive systems to be less robust. The potential benefits of stochastic resonance and external noise are also discussed with respect to processing subthreshold stimuli (e.g., Li, von Oertzen, & Lindenberger, 2006). We conclude by highlighting important challenges and outstanding research issues that remain to be answered in the study of IIV. PMID- 20025397 TI - Aging, time scales, and sensorimotor variability. AB - It is well established that there is an increased amount of intraindividual variability with aging in a variety of behavioral contexts. Here, we elaborate from a self-organization and dynamic systems framework to investigate the relevant time scales of variability as a function of aging and their relation to the changes in the amount and structure (frequency and time domains) of movement and postural variability. In particular, we examine evidence for the general hypotheses that (a) there is a reduction or even loss of shorter time scales in the control of movement with aging and (b) the shorter the time scale in motor output variability, the more sensitive the measure is as a biomarker to revealing the onset and early influence of aging and disease. The dynamic analysis of the time scales of variability distinguishes the distinctive roles of stability and noise in the increased amount of intraindividual variability with aging. PMID- 20025398 TI - Interindividual differences and intraindividual variability in the cortisol awakening response: an examination of age and gender. AB - This article shows age and gender differences in the magnitude and day-to-day variability of the cortisol awakening response (CAR) using a national sample of 1,143 adults who completed the second wave of the National Study of Daily Experiences, a part of the Midlife Development in the United States survey. Participants between the ages of 33 and 84 years completed 8 consecutive nightly interviews and provided 4 saliva samples (upon waking, 30 min after waking, before lunch, and before bed) on 4 consecutive interview days. Results revealed substantial day-to-day variability in the CAR as well as significant AgexGender interactions, indicating that although no systematic age-related differences emerged for women, the magnitude and day-to-day variability of the CAR increased with age among men. PMID- 20025400 TI - On the relation of mean reaction time and intraindividual reaction time variability. AB - Researchers often statistically control for means when examining individual or age-associated differences in variances, assuming that the relation between the 2 is linear and invariant within and across individuals and age groups. We tested this assumption in the domain of working memory by applying variance heterogeneity multilevel models to reaction times in the n-back task. Data are from the COGITO study, which comprises 101 younger and 103 older adults assessed in over 100 daily sessions. We found that relations between means and variances vary reliably across age groups and individuals, thereby contradicting the invariant linearity assumption. We argue that statistical control approaches need to be replaced by theoretical models that simultaneously estimate central tendency and dispersion of latencies and accuracies and illustrate this claim by applying the diffusion model to the same data. Finally, we note that differences in reliability between estimates for means and variances need to be considered when comparing their unique contributions to developmental outcomes. PMID- 20025399 TI - Intraindividual change and variability in daily stress processes: findings from two measurement-burst diary studies. AB - There is little longitudinal information on aging-related changes in emotional responses to negative events. In the present article, we examined intraindividual change and variability in the within-person coupling of daily stress and negative affect using data from 2 measurement-burst daily diary studies. Three main findings emerged. First, average reactivity to daily stress increased longitudinally, and this increase was evident across most of the adult lifespan. Second, individual differences in emotional reactivity to daily stress exhibited long-term temporal stability, but this stability was greatest in midlife and decreased in old age. Third, reactivity to daily stress varied reliably within persons (across-time), with individuals exhibiting higher levels of reactivity during times when reporting high levels of global subject stress in the previous month. Taken together, the present results emphasize the importance of modeling dynamic psychosocial and aging processes that operate across different time scales for understanding age-related changes in daily stress processes. PMID- 20025401 TI - Issues in intraindividual variability: individual differences in equilibria and dynamics over multiple time scales. AB - This article addresses three issues germane to experimental design and statistical analysis of intraindividual variability such as the articles contained within this special section. First, the time scale of the measurement of a process can have profound effects on the outcome of analyses of the resulting time series. Measurement in time poses special problems in the design of experiments: the time scale of the measurements must be appropriate for the time scale of the process. Second, deterministic and stochastic models should be fit at the individual level and only at a second level should individual differences in parameters be modeled. Third, one must consider the possibility that nomothetic relations may be exposed by the invariance of covariance between latent variables rather than within a factor analytic measurement model. PMID- 20025402 TI - Intraindividual variability in positive and negative affect over 45 days: do older adults fluctuate less than young adults? AB - Opposing scenarios about age-related increases and decreases in intraindividual variability are found in the literature: Whereas accumulating evidence indicates that cognitive functioning is characterized by an age-related increase of short term variability, age-related decreases in variability could be expected in affective states on the basis of theories of emotion regulation and self development. We examined age differences in intraindividual variability of positive affect (PA) and negative affect (NA) and in contingencies among daily affect, daily stress, and daily events using up to 45 daily assessments of 18 young (20-30 years) and 19 older (70-80 years) adults. Whereas age groups differed little in average affect levels, older adults showed significantly less variability in PA and NA than young adults. Age differences accounted for greater variance in variability than personality factors. Multilevel modeling indicated that for young but not older adults, PA was higher (lower) on days with a positive (negative) event, and NA was lower on days with a positive event. There were no age differences in daily affect reactivity to appraised stress severity. PMID- 20025403 TI - Emotional reactivity across the adult life span: the cognitive pragmatics make a difference. AB - Previously, we found that during films about age-typical losses, older adults experienced greater sadness than young adults, whereas their physiological responses were just as large. In the present study, our goal was to replicate this finding and extend past work by examining the role of cognitive functioning in age differences in emotional reactivity. We measured the autonomic and subjective responses of 240 adults (age range=20 to 70) while they viewed films about age-typical losses from our previous work. Findings were fully supportive of our past work: The magnitude of subjective reactions to our films increased linearly over the adult years, whereas there were no age differences on the level of physiological reactivity. We also found that the subjective reactions of adults with high pragmatic intelligence were of moderate size independent of their own age or the age relevance of the emotion elicitor. In contrast, the subjective reactions of adults low on pragmatic intelligence were more variable. Together, this evidence suggests that research on age differences in emotional reactivity may benefit from a perspective that considers individual difference variables as well as contextual variations. PMID- 20025404 TI - Effects of aging on experimentally instructed detached reappraisal, positive reappraisal, and emotional behavior suppression. AB - Emotion regulation includes multiple strategies that rely on different underlying abilities and that may be affected differently by aging. We assessed young, middle-aged, and older adults' ability to implement 3 emotion regulation strategies (detached reappraisal, positive reappraisal, and behavior suppression) in a laboratory setting, using standardized emotional stimuli and a multimethod approach to assessing regulation success. Results revealed age-related decline in ability to implement detached reappraisal, enhancement of ability to implement positive reappraisal, and maintenance of ability to implement behavior suppression. We discuss these findings in terms of their implications for emotion theory and for promoting successful aging. PMID- 20025405 TI - Cognitive aging and the adaptive use of recognition in decision making. AB - The recognition heuristic, which predicts that a recognized object scores higher on some criterion than an unrecognized one, is a simple inference strategy and thus an attractive mental tool for making inferences with limited cognitive resources--for instance, in old age. In spite of its simplicity, the recognition heuristic might be negatively affected in old age by too much knowledge, inaccurate memory, or deficits in its adaptive use. Across 2 studies, we investigated the impact of cognitive aging on the applicability, accuracy, and adaptive use of the recognition heuristic. Our results show that (a) young and old adults' recognition knowledge was an equally useful cue for making inferences about the world; (b) as with young adults, old adults adjusted their use of the recognition heuristic between environments with high and low recognition validities; and (c) old adults, however, showed constraints in their ability to adaptively suspend the recognition heuristic on specific items. Measures of fluid intelligence mediated these age-related constraints. PMID- 20025406 TI - Aging and integration of contingency evidence in causal judgment. AB - Age differences in causal judgment are consistently greater for preventative/negative relationships than for generative/positive relationships. In this study, a feature analytic procedure (Mandel & Lehman, 1998) was used to determine whether this effect might be due to differences in young and older adults' integration of contingency evidence during causal induction. To reduce the impact of age-related changes in learning/memory, the authors presented contingency evidence for preventative, noncontingent, and generative relationships in summary form; the meaningfulness of causal context was varied to induce participants to integrate greater or lesser amounts of this evidence. Young adults showed greater flexibility in their integration processes than did older adults. In an abstract causal context, there were no age differences in causal judgment or integration, but in meaningful contexts, young adults' judgments for preventative relationships were more accurate than older adults' and young adults assigned more weight to the contingency evidence confirming these relationships. These differences were mediated by age-related changes in processing speed. The decline in this basic cognitive resource may place boundaries on the amount or type of evidence that older adults can integrate for causal judgment. PMID- 20025407 TI - Aging and work: how do SOC strategies contribute to job performance across adulthood? AB - The authors examined the impacts of selection, optimization, and compensation (SOC) strategies-elective selection, loss-based selection, optimization, and compensation-on job performance across adulthood. A cross-sectional survey (Study 1, N=355) and a 5-day experience sampling study (Study 2, N=87) were conducted to assess Chinese insurance sales workers' global and momentary employment of SOC strategies at work and compare the effectiveness of these strategies in predicting their job performance. Study 1 revealed that the use of compensation predicted higher performance maintenance among older workers, whereas the use of elective selection contributed positively to sales productivity for both age groups, with stronger association for younger workers. Study 2 demonstrated that the positive impact of SOC strategies on global and momentary measures of job performance differed across tasks with various difficulty levels. When the task was perceived as highly difficult, older workers' greater use of elective selection predicted higher self-rated task performance; however, the positive association was weaker among younger workers. Older workers' greater use of the 4 SOC strategies was positively associated with sales increases when the task was not difficult or moderately difficult, yet the relationship was negative when the task was highly difficult. A reverse pattern was observed among younger workers. This article contributes to the understanding of working adults' psychological adaptation to the process of aging and reveals the moderating role of task difficulty on the association between SOC strategies and performance outcomes. PMID- 20025408 TI - Perceptions of aging across 26 cultures and their culture-level associates. AB - College students (N=3,435) in 26 cultures reported their perceptions of age related changes in physical, cognitive, and socioemotional areas of functioning and rated societal views of aging within their culture. There was widespread cross-cultural consensus regarding the expected direction of aging trajectories with (a) perceived declines in societal views of aging, physical attractiveness, the ability to perform everyday tasks, and new learning; (b) perceived increases in wisdom, knowledge, and received respect; and (c) perceived stability in family authority and life satisfaction. Cross-cultural variations in aging perceptions were associated with culture-level indicators of population aging, education levels, values, and national character stereotypes. These associations were stronger for societal views on aging and perceptions of socioemotional changes than for perceptions of physical and cognitive changes. A consideration of culture-level variables also suggested that previously reported differences in aging perceptions between Asian and Western countries may be related to differences in population structure. PMID- 20025409 TI - Effects of caregiver burden and satisfaction on affect of older end-stage renal disease patients and their spouses. AB - We examined the extent to which a 2-factor model of affect explains how the burdens and satisfactions experienced by caregivers influence their own well being and that of the spouses for whom they provide care. Using data from 315 older patients with end-stage renal disease and their spouses, we extended tests of Lawton et al.'s (1991) 2-factor model both longitudinally and dyadically. Multilevel modeling analyses partially support the 2-factor model. Consistent with the model, mean caregiver burden has a stronger effect on both caregiver and patient negative affect than does mean caregiver satisfaction. Contrary to the model, mean caregiver satisfaction has an effect on caregiver positive affect that is similar to that of mean caregiver burden, and it has no effect on patient positive affect. Time-varying effects of caregiver burden are consistent with the 2-factor model for caregiver but not patient negative affect. Time-varying effects of caregiver satisfaction are not consistent with the 2-factor model for either patients or caregivers. Results highlight the powerful role of caregiver burden for both caregivers and patients and suggest important new directions for conducting health-related research with late-life marital dyads. PMID- 20025410 TI - Working memory and aging: separating the effects of content and context. AB - In 3 experiments, we investigated the hypothesis that age-related differences in working memory might be due to the inability to bind content with context. Participants were required to find a repeating stimulus within a single series (no context memory required) or within multiple series (necessitating memory for context). Response time and accuracy were examined in 2 task domains: verbal and visuospatial. Binding content with context led to longer processing time and poorer accuracy in both age groups, even when working memory load was held constant. Although older adults were overall slower and less accurate than young adults, the need for context memory did not differentially affect their performance. It is therefore unlikely that age differences in working memory are due to specific age-related problems with content-with-context binding. PMID- 20025411 TI - Elevated false recollection of emotional pictures in young and older adults. AB - Current theories predict opposing effects of emotionally arousing information on false memory. If emotion enhances true recollection, then false recollection might be lower for emotional than for neutral pictures. However, if emotion enhances conceptual relatedness, then false recollection might increase for nonstudied but emotionally related pictures. We contrasted these 2 factors in young and older adults, using the International Affective Pictures Systems set (Lang, Bradley, & Cuthbert, 2005). Although both age groups used recollection in our task, false recollection was greatest for emotional pictures, supporting a conceptual relatedness account. Finally, even after accuracy differences were controlled, age was related to high-confidence false recollection of emotional pictures. PMID- 20025412 TI - Use of gaze for real-time mood regulation: effects of age and attentional functioning. AB - Older adults show positive preferences in their gaze toward emotional faces, and such preferences appear to be activated when older adults are in bad moods. This suggests that age-related gaze preferences serve a mood regulatory role, but whether they actually function to improve mood over time has yet to be tested. We investigated links between fixation and mood change in younger and older adults, as well as the moderating role of attentional functioning. AgexFixationxAttentional Functioning interactions emerged such that older adults with better executive functioning were able to resist mood declines by showing positive gaze preferences. Implications for the function of age-related positive gaze preferences are discussed. PMID- 20025413 TI - Subconscious facial expression mimicry is preserved in older adulthood. AB - It has been suggested that despite explicit recognition difficulties, implicit processing of facial expressions may be preserved in older adulthood. To directly test this possibility, the authors used facial electromyography to assess older (N=40) and young (N=46) adults' mimicry responses to angry and happy facial expressions, which were presented subliminally via a backward masking technique. The results indicated that despite not consciously perceiving the facial emotion stimuli, both groups mimicked the angry and happy facial expressions. Implications for emotion recognition difficulties in late adulthood are discussed. PMID- 20025414 TI - Individual differences and reliability of paired associates learning in younger and older adults. AB - The authors modeled individual nonlinear trajectories of learning using structured latent growth curves based on an exponential function with 3 parameters: initial performance, learning rate, and asymptotic performance. The 3 parameters showed reliable individual differences and the between-parameter correlations indicated that participants with high learning rates recalled more items initially. The asymptotic performance was unrelated to the learning rate and the initial performance. In addition, age and speed of information processing were included in the analyses. Age mainly affected negatively the asymptotic and the initial performance whereas speed of information processing affected the learning rate positively. Reliability estimates based on 2 similar learning conditions were moderate overall. PMID- 20025415 TI - Detecting threat-related intentional actions of others: effects of image quality, response mode, and target cuing on vigilance. AB - Three experiments examined the vigilance performance of participants watching videos depicting intentional actions of an individual's hand reaching for and grasping an object--involving transporting or using either a gun or a hairdryer- in order to detect infrequent threat-related actions. Participants indicated detection of target actions either manually or by withholding response. They also rated their subjective mental workload before and after each vigilance task. Irrespective of response mode, the detection rate of intentional threats declined over time on task and subjective workload increased, but only under visually degraded viewing conditions. This vigilance decrement was attenuated by temporal cues that were 75% valid in predicting a subsequent target action and eliminated with 100% valid cues. The findings indicate that detection of biological motion targets, and threat-related intentional actions in particular, although not attention sensitive under normal viewing conditions, is subject to vigilance decrement under degraded viewing conditions. The results are compatible with the view that the decrement in detecting threat-related intentional actions reflects increasing failure of attention allocation processes over time. PMID- 20025416 TI - Improving memory after interruption: exploiting soft constraints and manipulating information access cost. AB - Forgetting what one was doing prior to interruption is an everyday problem. The recent soft constraints hypothesis (Gray, Sims, Fu, & Schoelles, 2006) emphasizes the strategic adaptation of information processing strategy to the task environment. It predicts that increasing information access cost (IAC: the time, and physical and mental effort involved in accessing information) encourages a more memory-intensive strategy. Like interruptions, access costs are also intrinsic to most work environments, such as when opening documents and e-mails. Three experiments investigated whether increasing IAC during a simple copying task can be an effective method for reducing forgetting following interruption. IAC was designated Low (all information permanently visible), Medium (a mouse movement to uncover target information), or High (an additional few seconds to uncover such information). Experiment 1 found that recall improved across all three levels of IAC. Subsequent experiments found that High IAC facilitated resumption after interruption, particularly when interruption occurred on half of all trials (Experiment 2), and improved prospective memory following two different interrupting tasks, even when one involved the disruptive effect of using the same type of resource as the primary task (Experiment 3). The improvement of memory after interruption with increased IAC supports the prediction of the soft constraints hypothesis. The main disadvantage of a high access cost was a reduction in speed of task completion. The practicality of manipulating IAC as a design method for inducing a memory-intensive strategy to protect against forgetting is discussed. PMID- 20025417 TI - Using standards to improve middle school students' accuracy at evaluating the quality of their recall. AB - When recalling key term definitions from class materials, students may recall entirely incorrect definitions, yet will often claim that these commission errors are entirely correct; that is, they are overconfident in the quality of their recall responses. We investigated whether this overconfidence could be reduced by providing various standards to middle school students as they evaluated their recall responses. Students studied key term definitions, attempted to recall each one, and then were asked to score the quality of their recall. In Experiment 1, they evaluated their recall responses by rating each response as fully correct, partially correct, or incorrect. Most important, as they evaluated a particular response, it was presented either alone (i.e., without a standard) or with the correct definition present. Providing this full-definition standard reduced overconfidence in commission errors: Students assigned full or partial credit to 73% of their commission errors when they received no standard, whereas they assigned credit to only 44% of these errors when receiving the full-definition standard. In Experiment 2, a new standard was introduced: Idea units from each definition were presented, and students indicated whether each idea unit was in their response. After making these idea-unit judgments, the students then evaluated the quality of their entire response. Idea-unit standards further reduced overconfidence. Thus, although middle school students are overconfident in evaluating the quality of their recall responses, using standards substantially reduces this overconfidence and promises to improve the efficacy of their self-regulated learning. PMID- 20025418 TI - Effects of differential feedback on students' examination performance. AB - The effects of feedback on performance and factors associated with it were examined in a large introductory psychology course. The experiment involved college students (N = 464) working on an essay examination under 3 conditions: no feedback, detailed feedback that was perceived by participants to be provided by the course instructor, and detailed feedback that was perceived by participants to be computer generated. Additionally, these conditions were crossed with factors of grade (receiving a numerical grade or not) and praise (receiving a statement of praise or not). The task under consideration was a single-question essay examination administered at the beginning of the course. Detailed feedback on the essay, specific to individual's work, was found to be strongly related to student improvement in essay scores, with the influence of grades and praise being more complex. Generally, receipt of a tentative grade depressed performance, although this effect was ameliorated if accompanied by a statement of praise. Overall, detailed, descriptive feedback was found to be most effective when given alone, unaccompanied by grades or praise. It was also found that the perceived source of the feedback (the computer or the instructor) had little impact on the results. These findings are consistent with the research literature showing that descriptive feedback, which conveys information on how one performs the task and details ways to overcome difficulties, is far more effective than evaluative feedback, which simply informs students about how well they did. PMID- 20025419 TI - Responses to deceleration during car following: roles of optic flow, warnings, expectations, and interruptions. AB - More than 25% of accidents are rear-end collisions. It is essential to identify the factors that contribute to such collisions. One such factor is a driver's ability to respond to the deceleration of the car ahead. In Experiment 1, we measured effects of optic flow information and discrete visual and auditory warnings (brake lights, tones) on responses to deceleration during car following. With computer simulations of car-following scenes, university students pressed a button when the lead car decelerated. Both classes of information affected responses. Observers relied on discrete warnings when optic flow information was relatively less effective as determined by the lead car's headway and deceleration rate. This is consistent with DeLucia's (2008) conceptual framework of space perception that emphasized the importance of viewing distance and motion (and task). In Experiment 2, we measured responses to deceleration after a visual interruption. Scenes were designed to tease apart the role of expectations and optic flow. Responses mostly were consistent with optic flow information presented after the interruption rather than with putative mental expectations that were set up by the lead car's motion prior to the interruption. The theoretical implication of the present results is that responses to deceleration are based on multiple sources of information, including optical size, optical expansion rate and tau, and discrete warnings that are independent of optic flow. The practical implication is that in-vehicle collision-avoidance warning systems may be more useful when optic flow is less effective (e.g., slow deceleration rates), implicating a role for adaptive collision-warning systems. PMID- 20025420 TI - My imagination versus your feelings: can personal affective forecasts be improved by knowing other peoples' emotions? AB - A proposed remedy for biased affective forecasts is to base judgments on the actual feelings of people (surrogates) currently experiencing the event, rather than using imagination which conjures an inaccurate vision of the future. Gilbert et al. (2009) forced people to use surrogate reports by withholding all event information, resulting in better predictions. However, in life surrogate information rarely supplants event information--can people effectively integrate both types of information into their judgments? In five studies, respondents predicted the impact of a health state on their own happiness. Respondents incorporated surrogate information into their judgments both in the presence and absence of event information. However, they inappropriately discounted other people's experiences as a valid predictor of their own--particularly in the presence of event information--and imagined their happiness would be different to surrogates' happiness. Excluding preexisting event knowledge, changing the size of the surrogate sample, or increasing the size of the response scale did not alter the adjustment. Although surrogate information improved affective forecasts, its influence was diminished by the presence of event information. PMID- 20025421 TI - Human figure drawings and children's recall of touching. AB - In 2 studies, children ages 3 to 7 years were asked to recall a series of touches that occurred during a previous staged event. The recall interview took place 1 week after the event in Study 1 and immediately after the event in Study 2. Each recall interview had 2 sections: In 1 section, children were given human figure drawings (HFDs) and were asked to show where the touching took place; in the other section, the same questions were asked without the HFDs (verbal condition). Children were randomly assigned to 2 different conditions: HFD 1st/verbal 2nd or verbal 1st/HFD 2nd. There were 2 major findings. First, HFDs elicited more errors than the verbal condition when used to probe for information that the child had already been asked. Second, regardless of interview method, children had poor recall of the touches even when these occurred minutes before the interview. It is suggested that cognitive mechanisms involving memory and semantics underlie children's poor recall of touching in both verbal and HFD conditions. PMID- 20025422 TI - Predicting sentencing for low-level crimes: comparing models of human judgment. AB - Laws and guidelines regulating legal decision making are often imposed without taking the cognitive processes of the legal decision maker into account. In the case of sentencing, this raises the question of whether the sentencing decisions of prosecutors and judges are consistent with legal policy. Especially in handling low-level crimes, legal personnel suffer from high case loads and time pressure, which can make it difficult to comply with the often complex rulings of the law. To understand the cognitive processes underlying sentencing decisions, an analysis of trial records in cases of larceny, fraud, and forgery was conducted. Applying a Bayesian approach, five models of human judgment were tested against each other to predict the sentencing recommendations of the prosecution and to identify the crucial factors influencing sentencing decisions. The factors influencing sentencing were broadly consistent with the penal code. However, the prosecutors considered only a limited number of factors and neglected factors that were legally relevant and rated as highly important. Furthermore, testing the various cognitive judgment models against each other revealed that the sentencing process was apparently not consistent with the judgment policy recommended by the legal literature. Instead, the results show that prosecutors' sentencing recommendations were best described by the mapping model, a heuristic model of quantitative estimation. According to this model, sentencing recommendations rely on a categorization of cases based on the cases' characteristics. PMID- 20025423 TI - Introduction. PMID- 20025424 TI - Programmatic research at a specialty psychiatric inpatient clinic: Opportunities, challenges, and future directions. PMID- 20025425 TI - Integrating outcomes assessment and research into clinical care in inpatient adult psychiatric treatment. AB - The authors describe an evolving outcomes project implemented across the adult inpatient programs at The Menninger Clinic. In the inpatient phase of the project, patients complete a computerized battery of standardized scales at admission, at biweekly intervals throughout treatment, and at discharge. In addition to providing aggregate data for outcomes research, these assessments are incorporated into routine clinical care, with results of each individual assessment provided to the treatment team and to the patient. The inpatient phase of the project employs Web-based software in preparation for a forthcoming follow up phase in which patients will continue after discharge to complete assessments on the same computer platform. This article begins with a brief overview of related research at the Clinic to place the current project in local historical context. Then the authors describe the assessment instruments, the ways in which the assessments are integrated into clinical care, plans for follow-up assessments, the central role of information technology in the development and implementation of the project, the primary research questions, and some of the major challenges in implementing the project. The article concludes with a discussion of the ways in which the project can serve as a platform for a broad future research agenda. PMID- 20025426 TI - Psychosocial characteristics of psychiatric inpatients at admissions and discharge: The Menninger Clinic Adult Outcomes Project. AB - The authors present a first look at the data from The Menninger Clinic Adult Outcomes Project. They provide descriptive data from 443 patients admitted to the Clinic between April 2008 and May 2009. Patients show significant improvement on a range of standardized measures from admission to discharge (effect sizes range from moderate to large, 0.31 to 1.44), and there are some differences among clinical programs within the hospital in the extent of change on some scales. A comparison of patients who did and did not complete discharge assessments showed minimal differences between groups on admissions variables. These results attest to the substantial impact of relatively long-term intensive inpatient treatment on improving clinical symptoms and functioning, but further studies are needed to determine the trajectory of change in the hospital and, most importantly, at follow-up. PMID- 20025427 TI - The development of a mentalization-based outcomes and research protocol for an adolescent inpatient unit. AB - The authors describe the development of a theory-driven assessment and research protocol at the Adolescent Treatment Program of The Menninger Clinic. First, the theoretical framework behind a mentalization-based model for assessment and treatment is described. Next, the process whereby measures were selected to operationalize key components of the mentalization-based model is discussed, including a brief discussion of each measure and assessment procedure. The next section describes the clinical and research use of the data collected. Here, the authors describe how outcomes assessment information is integrated into the clinical decision-making process, and they outline the research questions they aim to answer through the assessment protocol. The authors conclude with a section on the challenges, pitfalls, and future directions of the project. PMID- 20025428 TI - Implementing an evidence-based approach to working with suicidal inpatients. AB - In this article, the authors describe the implementation of a new approach to working with suicidal patients in an inpatient psychiatric facility. The Collaborative Assessment and Management of Suicidality (CAMS) has been under development for almost two decades, but has rarely been implemented or studied in an inpatient setting. Here the authors describe the context for this project, the nature of the CAMS intervention in this setting, the structure for the research aspect of the project, and some of the implementation issues that have arisen as the project has unfolded. The authors conclude that a solid foundation has been laid for an initiative that will both enhance assessment and treatment of at-risk patients and contribute to a body of knowledge that is currently lacking in evidence for interventions with suicidal patients. PMID- 20025429 TI - Evidence-based practice and research scholars programs: Supporting excellence in psychiatric nursing. AB - Delivery of quality psychiatric nursing care depends, to a certain extent, on nurses' ability to apply sound evidence to practice. Much of the needed evidence is in the form of research. The term evidence-based practice (EBP) is used to describe the application of research and other forms of clinically relevant information to practice. This article provides a historical context for EBP and describes two programs at The Menninger Clinic that support nurses on their journey to excellence and directly impact the quality of nursing care delivered at Menninger. The Evidence-based Practice Scholars Program and the Research Scholars Program are mission critical initiatives at the hospital. The Evidence based Practice Scholars Program has led to the implementation of EBP policies and guidelines. The first project of the Research Scholars Program has been launched. The results of the program have the potential to improve the quality of nursing care as the current study will inform nurses about patients' perceptions of good nursing care and how these perceptions correspond with professional standards and scope of practice. Whether nurses are interested in being consumers of research or involved in the conduct of research, programs such as these help create a research-minded culture, much needed in contemporary health care arenas. These programs have the potential to transform nursing practice, improve the quality of nursing care, and enhance nursing job satisfaction. These are essential elements needed to sustain an environment that fosters excellence. In an era where patients and providers demand the application of research to health care, organizations must respond by investing in programs to promote excellence in this regard. PMID- 20025431 TI - Handgrip strength of the elderly after hip fracture repair correlates with functional outcome. AB - PURPOSE: To investigate the relationship between handgrip strength (HG) and upper limb functioning of hip fracture operated elderly patients and their functional outcome 6 months post-op. METHODS: Retrospectively, data on 105 post-operative hip fracture patients was analyzed. Functional level was measured by the Functional Independence Measure (FIM) and HG by hand dynamometer, 7-10 days, 1, 3 and 6 months post-op. FIM1 up-to FIM4 (FIM1-FIM4) and HG1 up-to HG4 (HG1-HG4) were determined. Pre-fracture upper limb function was evaluated using the disabilities of the arm, shoulder and hand (DASH) questionnaire. The Mini-Mental State Examination evaluated cognition. RESULTS: FIM1-FIM4 scores increased during the 6-month follow-up; HG1-HG4 scores remained unchanged. No differences were found in FIM1-FIM4 and DASH scores between genders. HG1-HG4 scores were significantly higher in men (p = 0.04-0.005). FIM1-FIM4, HG1-HG4 and DASH scores were higher in the cognitively normal patients (p < 0.001). A fair correlation was found between all HG and FIM scores (R = 0.36-0.71, p < 0.001), and between DASH and FIM scores (R = 0.5-0.7, p < 0.001). Log HG1 and FIM4 scores were highly correlated (R(2) = 0.54, p < 0.001). Regression analysis found that only, and in the following order were Log HG1, FIM1, DASH and age significant factors affect FIM4 score (R(2) = 0.69). CONCLUSIONS: HG and FIM scores, a week post-op., combined with upper limb functioning and age, can have a 69% prediction accuracy for motor functioning 6 months post-fracture. HG measurement, a week post-hip fracture repair, might be considered in estimating functional motor outcome, and eventually additional interventions should be employed to increase muscle strength and, thus, improve functional outcome. PMID- 20025432 TI - Inhalation exposure systems for the development of rodent models of sulfur mustard-induced pulmonary injury. AB - Sulfur mustard (SM) is a chemical threat agent for which its effects have no current treatment. Due to the ease of synthesis and dispersal of this material, the need to develop therapeutics is evident. The present manuscript details the techniques used to develop SM laboratory exposure systems for the development of animal models of pulmonary injury. These models are critical for evaluating SM injury and developing therapeutics against that injury. Iterative trials were conducted to optimize a lung injury model. The resulting pathology was used as a guide, with a goal of effecting homogeneous and diffuse lung injury comparable to that of human injury. Inhalation exposures were conducted by either nose-only inhalation or intubated inhalation. The exposures were conducted to either directly vaporized SM or SM that was nebulized from an ethanol solution. Inhalation of SM by nose-only inhalation resulted in severe nasal epithelial degeneration and minimal lung injury. The reactivity of SM did not permit it to transit past the upper airways to promote lower airway injury. Intratracheal inhalation of SM vapors at a concentration of 5400 mg x min/m(3) resulted in homogeneous lung injury with no nasal degeneration. PMID- 20025433 TI - Ocular ciprofloxacin hydrochloride mucoadhesive chitosan-coated liposomes. AB - The aim of this work is to improve the ocular bioavailability of ciprofloxacin hydrochloride (CPX) through the preparation of ocular mucoadhesive chitosan (CS) coated liposomes. Liposomes were prepared by the thin film hydration technique, using different molar ratios of L-alpha-phosphatidylcholine (PC), cholesterol (CH), stearylamine (SA) and dicetyl phosphate (DP). CS was used to coat the optimal liposomal formulae. The prepared formulae were characterized regarding encapsulation efficiency (%EE), particle size, physical morphology and in vitro drug release. The in vivo characterization of the prepared formulae was performed through evaluating the level of CPX in the external eye tissue of nine albino rabbits. Results showed an alteration in release rate and %EE of CPX from liposomal formulae upon varying the molar ratios of the lipid bilayer composition. The optimal liposomal formulae F1 (10:0, PC:CH), F12 (10:0:0.5, PC:CH:SA) and F15 (10:0:1, PC:CH:DP), showed % EE of 38.5 +/- 2.10, 39.65 +/- 1.85 and 30.05 +/- 0.75 and % in vitro release after 8 hours (Q(8h)) of 78.15 +/- 2.4, 54.07 +/- 2.3 and 62.14 +/- 2.9, respectively. In vitro drug release and in vivo results confirmed that CS-coated liposomal formulae have exhibited a higher retention of CPX. Consequently, CS-coated liposomes could be a promising approach to increase the ocular bioavailability of CPX. PMID- 20025434 TI - Impact of biological agents and tissue engineering approaches on the treatment of rheumatic diseases. AB - The treatment of rheumatic diseases has been the focus of many clinical studies aiming to achieve the best combination of drugs for symptom reduction. Although improved understanding of the pathophysiology of rheumatic diseases has led to the identification of effective therapeutic strategies, its cure remains unknown. Biological agents are a breakthrough in the treatment of these diseases. They proved to be more effective than the other conventional therapies in refractory inflammatory rheumatic diseases. Among them, tumor necrosis factor inhibitors are widely used, namely Etanercept, Infliximab, or Adalimumab, alone or in combination with disease-modifying antirheumatic drugs. Nevertheless, severe adverse effects have been detected in patients with history of recurrent infections, including cardiac failure or malignancy. Currently, most of the available therapies for rheumatic diseases do not have sufficient tissue specificity. Consequently, high drug doses must be administrated systemically, leading to adverse side effects associated with its possible toxicity. Drug delivery systems, by its targeted nature, are excellent solutions to overcome this problem. In this review, we will describe the state-of-the-art in clinical studies on the treatment of rheumatic diseases, emphasizing the use of biological agents and target drug delivery systems. Some alternative novel strategies of regenerative medicine and its implications for rheumatic diseases will also be discussed. PMID- 20025435 TI - Alcohol and aldehyde dehydrogenase polymorphisms in Chinese and Indian populations. AB - The association between two functional polymorphisms in alcohol dehydrogenase (ADH2/ADH1B) and aldehyde dehydrogenase (ALDH2) genes and alcohol dependence was examined in 182 Chinese and Indian patients undergoing treatment for alcohol dependence and 184 screened control subjects from Singapore. All subjects were screened by the Alcohol Use Disorders Identification Test (AUDIT). Patients were also administered the Severity of Alcohol Dependence Questionnaire (SADQ). Polymorphisms were genotyped by allele-specific polymerase chain reaction and selected genotypes confirmed by DNA sequencing or restriction fragment length polymorphism. Our results showed that frequencies of ADH1B*2 and ALDH2*2 were higher in controls compared to alcohol-dependent subjects for both Chinese and Indians. Frequencies of these two alleles were also higher in the 104 Chinese controls compared to the 80 Indian controls. None of the eight Chinese who were homozygous for both protective alleles was alcohol dependent. The higher frequencies of the protective alleles could explain the lower rate of alcohol dependence in Chinese. PMID- 20025436 TI - Decreased psychological well-being in late 'chronotypes' is mediated by smoking and alcohol consumption. AB - Individuals are different 'chronotypes' with early 'larks' and late 'owls' forming the limits of a normal distribution in the population. We recently described that late chronotypes who suffer from a conflict between internal and external time ('social jetlag') suffer from more mental distress and are more likely to smoke than early chronotypes (Wittmann, Dinich, Merrow, and Roenneberg, 2006 . Social jetlag: mis-alignment of biological and social time. Chronobiology International, 23:497-509.). We performed a detailed analysis of the same database collected in 2002 comprising 134 daily smokers and 366 nonsmokers, scrutinizing the relationships between chronotype, smoking, and alcohol consumption as well as psychological well-being using a multiple mediation analysis. On average, smokers tend to be later chronotypes, report more sleep associated psychosomatic symptoms, are more depressed, less balanced, and less vigilant. The mediation analysis suggests that only those late chronotypes who smoke and those who drink more suffer from increased psychological distress. We suggest that 'chronotype' is introduced as an additional factor in substance use, that is, when considering motives for smoking and drinking. PMID- 20025437 TI - "Adderall is definitely not a drug": justifications for the illegal use of ADHD stimulants. AB - In-depth interviews were conducted in 2007 with 175 undergraduate students (94 males, 81 females, 13 non-Caucasian) at a large, public southeastern research university located in an urban area in the United States. Our primary goal was to identify how these students conceive of Attention Deficit Hyperactivity Disorder (ADHD) stimulants and their illegal use. We discovered that these students frame stimulant use as both physically harmless and morally acceptable. Specifically, these students justify their drug use through the use of four recurring prostimulant arguments: 1) comparison-and-contrast, 2) all-things-in-moderation, 3) self-medicating, and 4) minimization arguments. We discuss limitations to the study and conclude by suggesting five strategies for prevention researchers that would directly target these four arguments. PMID- 20025438 TI - The development and psychometric properties of a new measure of perceived stigma toward substance users. AB - A self-report measure of perceived stigma toward substance users was developed and studied. An initial measure was created based on a previously developed scale that was rated by experts for content validity and quality of items. The scale, along with other measures, was administered to 252 people in treatment for substance problems in the United States during 2006-2007. Refinement efforts resulted in an eight-item scale with good face validity, construct validity, and adequate levels of internal consistency. Most relationships with other constructs were as expected. Findings suggest that perceived stigma is distinct from other forms of stigma. PMID- 20025439 TI - Knowledge about Hepatitis-C among methadone maintenance treatment patients in Israel. AB - Ignorance about Hepatitis-C (HCV) among drug users, treatment staff, and policy makers thwarts treatment uptake and facilitates virus transmission. We assessed knowledge about HCV among methadone patients in Israel, where effective HCV treatment is provided at low-cost within the national health insurance framework, yet few infected methadone patients are treated. In 2006, 512 patients in two methadone clinics in Israel were interviewed, of whom 53% were HCV-positive. The clinics were purposively selected from the 11 methadone clinics in the country. Respondents exhibited poor knowledge about HCV, particularly about diagnosis and treatment. Lesser-educated respondents were three times more likely to score low on HCV-knowledge compared to those with 12+ years of schooling (AOR = 2.97, 95% CI = 1.5-5.7. HCV-negative patients were also three-times more likely than HCV positive patients to score low on the HCV-knowledge scale (Adjusted Odds Ratio = 3.0, 95% Confidence Interval = 1.9-4.7). Enhancing HCV-knowledge may help patients avoid becoming infected and infecting others, allay exaggerated fears about hepatitis, and facilitate HCV-treatment initiation among those infected. PMID- 20025440 TI - Communication between researchers and practitioners: findings from a qualitative evaluation of a large-scale college intervention. AB - Many community-based intervention studies experience problems with collaboration between researchers and practitioners. A preferred strategy appears to be to form community coalitions to carry out the proposed interventions in the community, but doing so risks shifting the focus from intervention objectives to coalition process. As a by-product, coalitions often lack understanding of the project goals and are not given specific instructions on how to implement the intervention. In contrast to conventional wisdom, the Safer California Universities study implemented a very directive approach in collaboration with local liaisons on the participating campuses, even though this approach is seen to risk cooperation or commitment from collaborators. This paper reports on the findings of a qualitative study based on interviews with campus liaisons of how the directive approach was perceived on the participating campuses. Findings indicate that the strategy was successful in terms of "getting things done" but could have been improved in terms of liaison involvement in setting objectives, and in finding the optimal level of specificity. PMID- 20025441 TI - Exploring survey participation, data combination, and research validity in a substance use study: an application of hierarchical linear modeling. AB - A sound decision regarding combination of datasets is critical for research validity. Data were collected between 1996 and 2000 via a 99-item survey of substance use behaviors. Two groups of 7th-12th grade students in predominately White communities are compared: 166,578 students from 193 communities with high survey participation and 41,259 students from 65 communities with lower participation. Hierarchical logistic models are used to explore whether the two datasets may be combined for further study of community-level substance use effects. "Scenario analysis" is introduced. Results suggest the datasets may reasonably be combined. Limitations and further research are discussed. PMID- 20025442 TI - Binge use and sex and drug use behaviors among HIV(-), heterosexual methamphetamine users in San Diego. AB - This study identified sociodemographic factors, drug using practices, sexual behaviors, and motivational factors associated with binge (a period of uninterrupted) methamphetamine (MA) use among heterosexual MA users. SAMPLE AND METHOD: The FASTLANE study provided cross-sectional data collected by audio computer-assisted self-interview (ACASI) between June 2001 and August 2004 from 451 HIV-negative MA users in San Diego, California, USA who had engaged in unprotected sex and used MA in the previous two months. RESULTS: The study sample was 67.8% male, 49.4% Caucasian, 26.8% African-American, and 12.8% Hispanic with a mean age of 36.6 years; 183 (40.5%) reported binge use in the past 2 months. Compared with non-binge users, binge users of MA were more likely to report risky drug use and sex behaviors and differed in motivations to initiate and currently use MA. The final logistic regression model for binge use included more days of MA use in the last month, ever treated for MA use, injection drug use, higher Beck Depression Inventory score, "experimentation" as a motivation for initiating MA use, and engaging in sex marathons while high on MA. HIV prevention efforts should differentiate and address these differences in motivations for MA use and the associated HIV-risk sex and drug use behaviors as key targets for effective intervention. PMID- 20025443 TI - A focus-group study on spirituality and substance-user treatment. AB - Focus groups were conducted in 2005-2006 with 25 urban methadone-maintained outpatients to examine beliefs about the role of spirituality in addiction and its appropriateness in formal treatment. Thematic analyses suggested that spirituality and religious practices suffered in complex ways during active addiction, but went "hand in hand" with recovery. Participants agreed that integration of a voluntary spiritual discussion group into formal treatment would be preferable to currently available alternatives. One limitation was that all participants identified as strongly spiritual. Studies of more diverse samples will help guide the development and evaluation of spiritually based interventions in formal treatment. PMID- 20025445 TI - Social support and non-safe use of licit substances by Greek medical students. AB - This study aims to investigate the relationship between social support and non safe use of nicotine and alcohol among medical students, in relation to other contributing factors. Data were collected during 2002-2003. Participants (N = 135), medical students at the Democritus University of Thrace, Alexandroupolis, Greece, were allocated into groups per sex according to their smoking and alcohol drinking patterns. Comparisons were made with respect to rating scores on measures assessing perceived social support, psychopathological problems, stressful life events, physical health, and satisfaction with life. Data were analyzed using chi-square, Mann-Whitney U test, and factor analysis. Female students' non-safe use of licit substances is positively related to higher scores on perceived social support and lower scores on psychopathological dimensions. Despite some limitations of this study, findings bring about implications on antismoking and antialcoholic campaigns. PMID- 20025444 TI - Can drug users be effective change agents? Yes, but much still needs to change. PMID- 20025446 TI - Longitudinal observation of a sample of German drug consumption facility clients. AB - INTRODUCTION: We aimed at investigating whether attendance of a drug consumption facility (DCF) was associated with both a reduction of drug-associated at-risk behavior and referral to the health care treatment system. METHODS: A sample of 129 consecutive clients out of those 256 who self-referred to the DCF during the 13-month observation period (i.e., from November 2002 to December 2003) was interviewed both at baseline and at 1-, 2-, 3-, and 6-month follow-ups. Subjects were repeatedly assessed using a structured approach based on both the European Addiction Severity Index (EuropASI) and the Deutsche Gesellschaft fuer Suchtforschung und Therapie (DG-Sucht). RESULTS: Typical DCF clients were males, in their early 30s, single, with no vocational training, and with a long history of injectable opiate addiction in the context of polydrug misuse. A recent discharge from prison was recorded in 37% of cases. Median length of DCF attendance was of 5 weeks; 22% of clients attended for less than 1 week. Although with respect to the period previous to recruitment at-risk behavior rates remained unchanged, by the 3-month follow-up 13 (10%) clients out of those 129 who had initially enrolled had taken advantage of the DCF counseling opportunities. Some 37% of clients were referred on to start a methadone treatment following their DCF experience. DISCUSSION: DCF attendance was not associated with reduction in at-risk behavior over time, but a need was here identified for additional intervention to be available in the DCF to address clients' psychosocial issues. Limitations of the present study include both issues related to the representativeness of the sample of clients here recruited and the lack of a control/comparison group. PMID- 20025447 TI - Adolescents binge drinking when on holiday: an evaluation of a community intervention based on self-regulation. AB - This paper presents a case study of a community intervention aimed at reducing excessive drinking in young men on holiday in seaside camping resorts in the Netherlands. The self-regulated voluntary covenant of parties concerned was evaluated on the basis of several types of data (all collected in 2004 ): questionnaires administered to young men (N = 191), observations carried out by trained "peers," nuisance questionnaires among city center residents (N = 121), and interviews with local actors. Compliance with measures as well as the effects of measures are discussed. Limitations are noted and recommendations for optimizing the potential of community interventions are made. PMID- 20025448 TI - HIV testing in hazardous drinking: a survey analysis. AB - Hazardous drinking increases the risk of contracting and transmitting human immunodeficiency virus (HIV). This study aimed to determine the prevalence of HIV testing among hazardous drinkers using the 2006 Behavioral Risk Factor Surveillance System dataset. Adjusted odds ratios were obtained using multivariable logistic regression for the relationship between hazardous drinking and HIV testing. Results indicated that adjusting for confounders, hazardous drinkers (1) had 0.86 (95% CI: 0.84, 0.89) the odds of ever having had an HIV test compared to nonhazardous drinkers. Therefore public health interventions targeted at hazardous drinkers to increase the rate of HIV testing are suggested. The study's limitations are noted. PMID- 20025449 TI - Adaptation and validation of a self-report measure to evaluate substance use among Sri Lankan adolescents. AB - This study translated and validated the model Student Questionnaire (SQ) among 13 to 18-year-old Sri Lankan adolescents. A systematic procedure was followed to translate the SQ into Sinhala language. The Sinhala version was titled Adolescent Substance Use Student Questionnaire (ASUSQ). A Delphi process was conducted to evaluate consensual validity of ASUSQ. Test-retest study (N = 120) ensured reliability. Contrasted group technique study (N = 200) among users and nonusers established construct validity. Data collection instruments were SQ, its Sinhala version, structured-interview schedule, and focus group discussion guide. Sinhala speaking students were randomly selected from schools in a semi-urban area. Applicability of the ASUSQ and further research are discussed. The study's limitations are noted. PMID- 20025450 TI - Causes and consequences of increasing club drug use in China: a descriptive assessment. AB - Club drugs have quickly become the most widespread "drugs of abuse" in China. Using data from a convenience sample of 730 club drug users in Shanghai in 2006 , we explored the causes and consequences of club drug use. Descriptive analyses suggest that club drug use is typically polydrug use. Polydrug use is strongly associated with weakened social control, drug use social influences, and a sensation-seeking personality; in addition, it is associated with more negative health and social consequences. Both polydrug and single-club-drug users are at high risk of sexually acquiring and/or transmitting HIV. The study's limitations are noted, and future research is suggested. PMID- 20025451 TI - Diversion and injection of methadone and buprenorphine among clients in public opioid treatment clinics in New South Wales, Australia. AB - A survey of 448 clients receiving opioid treatment in public clinics in Australia was conducted during 2005, exploring diversion and injection of supervised methadone and buprenorphine, frequency and reported effects of injecting, and the cost and availability of street-purchased pharmacotherapies. The rates of diversion in the preceding 12 months were over three times higher among participants receiving supervised buprenorphine (15.3%) than among those receiving supervised methadone (4.3%). While 26.5% of participants currently prescribed buprenorphine had ever injected buprenorphine, 65.9% of those prescribed methadone reported ever injecting methadone. The majority of participants did not appear to have extensive experience of injecting their medication and most expressed a preference for taking it as directed. Further research is required to determine the optimal approach for the supervised administration of buprenorphine that maximizes the benefits of treatment and minimizes harm and the risk of diversion. The study's limitations are noted. PMID- 20025452 TI - Alcohol use and physical health in adolescence: a general population survey of 8,983 young people in North-Trondelag, Norway (the Young-HUNT study). AB - AIMS: To investigate the relationship between adolescents' alcohol use and physical health. DESIGN: A total population, cross-sectional survey of adolescents, aged 13-19 years, attending secondary or high school in Nord Trondelag County, Norway. 8,983 youths (91%) answered the Young-Hunt questionnaire in the 1995-1997 HUNT-survey. METHOD: Self-reported data mainly analyzed by logistic regression models. Findings 80% of the respondents had tried alcohol, and 29% reported more than 10 intoxications. Ill health perception was associated with frequent alcohol intoxications. Tension symptoms had the closest relationship to frequency of intoxications in both genders. Girls that frequently used health services had frequent alcohol intoxications. FUNDING: Central Norway Regional Health and the County Council of Nord-Trondelag. CONCLUSIONS: There is a close association of physical health complaints and alcohol intoxication frequency in Norwegian teenagers. The study's limitations were noted. PMID- 20025453 TI - Challenges of reliability and validity in the identification and monitoring of emerging drug trends. AB - There is increasing pressure on drug monitoring systems to achieve earlier detection and greater precision in reporting of emerging drug use trends. Such systems typically operate in settings where government interest and the drug use trends themselves can be fluid. To achieve the goal of informing timely policy and practice responses in this environment, drug use monitoring systems must be flexible and responsive, as well as reliable and valid. This paper explores three interrelated areas relevant to trend monitoring that can benefit from a clearer focus in terms of increasing validity and reliability: the research paradigm to which systems adhere; the selection of sources or drug use indicators utilized by systems; and the process of analysis used by systems to ensure valid results. The reliability and validity of currently utilized drug use related indicators is discussed, with a focus on the validity of data sources as measures of emerging drug use trends. The relevance and utility of current descriptives such as "lagged" and "leading edge" indicators are assessed. Five dimensions, against which the validity of drug use indicators may be assessed in a trend-monitoring context are proposed as an alternative. Faced with a lack of clear conceptual frameworks underpinning and driving monitoring systems, it is argued that a pragmatic research paradigm can be adopted as a basis for guiding selection of indicators and helping to make explicit the concurrent or supplementary triangulation and analysis procedures on which valid results are necessarily founded. The current trend of using triangulation as the primary means of ensuring the validity of systems is critically reviewed and a challenge is issued to the field to make the analysis process more overt. No external funding was received for this article. PMID- 20025454 TI - Improved quality of life, clinical, and psychosocial outcomes among heroin dependent patients on ambulatory buprenorphine maintenance. AB - AIM: A prospective longitudinal design was employed to examine the effects of buprenorphine maintenance on quality of life (QOL), clinical, and psychosocial characteristics of heroin-dependent patients. METHOD: Between 2003 and 2005 data were collected on 259 patients attending the outpatient centers for treatment of drug addictions across Israel, of which 157 were reevaluated 16 weeks later and 105 reevaluated 32 weeks later using the Clinical Global Impression, Distress Scale for Adverse Symptoms, Quality of Life Enjoyment and Satisfaction Questionnaire, General Health Questionnaire, General Self-Efficacy Scale, and the Multidimensional Scale of Perceived Social Support. Univariate and multivariate analyses were conducted to examine the association between the parameters and the cross-sectional and longitudinal predictions of the QOL outcomes. RESULTS: The groups did not differ in baseline values and their post-treatment ratings revealed significant improvement on virtually all the scales. Perceived self efficacy and social support from friends and significant others at baseline as well as their changes over time were the best predictors of the QOL in the short and long terms. The study's limitations are noted. CONCLUSIONS: The beneficial effects on the QOL were associated with improvement in the psychosocial parameters and a reduction in buprenorphine-related side effects and psychological distress. This study could stimulate research to compare the QOL related to buprenorphine and methadone treatment and serve as a basis on which a controlled study should be performed. PMID- 20025455 TI - Strategies for articular cartilage lesion repair and functional restoration. AB - Injury of articular cartilage due to trauma or pathological conditions is the major cause of disability worldwide, especially in North America. The increasing number of patients suffering from joint-related conditions leads to a concomitant increase in the economic burden. In this review article, we focus on strategies to repair and replace knee joint cartilage, since knee-associated disabilities are more prevalent than any other joint. Because of inadequacies associated with widely used approaches, the orthopedic community has an increasing tendency to develop biological strategies, which include transplantation of autologous (i.e., mosaicplasty) or allogeneic osteochondral grafts, autologous chondrocytes (autologous chondrocyte transplantation), or tissue-engineered cartilage substitutes. Tissue-engineered cartilage constructs represent a highly promising treatment option for knee injury as they mimic the biomechanical environment of the native cartilage and have superior integration capabilities. Currently, a wide range of tissue-engineering-based strategies are established and investigated clinically as an alternative to the routinely used techniques (i.e., knee replacement and autologous chondrocyte transplantation). Tissue-engineering based strategies include implantation of autologous chondrocytes in combination with collagen I, collagen I/III (matrix-induced autologous chondrocyte implantation), HYAFF 11 (Hyalograft C), and fibrin glue (Tissucol) or implantation of minced cartilage in combination with copolymers of polyglycolic acid along with polycaprolactone (cartilage autograft implantation system), and fibrin glue (DeNovo NT graft). Tissue-engineered cartilage replacements show better clinical outcomes in the short term, and with advances that have been made in orthopedics they can be introduced arthroscopically in a minimally invasive fashion. Thus, the future is bright for this innovative approach to restore function. PMID- 20025456 TI - Bone marrow stem cells contribute to alcohol liver fibrosis in humans. AB - Bone marrow-derived stem cell (BMSC) contribution to liver repair varies considerably and recent evidence suggests these cells may contribute to liver fibrosis. We investigated the mobilization and hepatic recruitment of bone marrow (BM) stem cells in patients with alcohol liver injury and their contribution to parenchymal/non-parenchymal liver cell lineages. Liver biopsies from alcoholic hepatitis (AH) patients and male patients, who received a female liver transplant and developed AH, were analyzed for BM stem cell content by fluorescence in situ hybridization and immunostaining. Y chromosome analysis was performed, along with co-staining for hepatocyte, biliary, myofibroblast, and Ki-67 markers. Blood CD34(+) levels were quantified in AH patients by flow cytometry. AH patients had increased CD34(+) cell counts in liver tissue (1.834% +/- 0.605%; P < 0.05) and in blood (0.195% +/- 0.063%; P < 0.05) as compared with matched controls (0.299% + 0.208% and 0.067% +/- 0.01%). A proportion of hepatic myofibroblasts were BM derived (7.9%-26.8%) as deemed by the co-localization of Y chromosome/alpha smooth muscle actin (alpha-SMA) staining. In the cross-sex liver grafts with AH, 5.025% of the myofibroblasts were co-staining for CD34, suggesting that a population of CD34(+) cells were contributing to the hepatic myofibroblast population. There was no evidence of BM contribution to hepatocyte or biliary cell differentiation, nor evidence of increased hepatocyte regeneration. Alcohol liver injury mobilizes CD34(+) stem cells into the circulation and recruits them into the liver. These BMSCs contribute to the hepatic myofibroblast population but not to parenchymal lineages and do not promote hepatocyte repair. PMID- 20025457 TI - The right idea at the right time and writing Renku with us at Nanomedicine. PMID- 20025458 TI - Nanomedicine-based cancer targeting: a new weapon in an old war. PMID- 20025460 TI - Efavirenz-loaded polymeric micelles for pediatric anti-HIV pharmacotherapy with significantly higher oral bioavailability [corrected]. AB - Children constitute the most challenging population in anti-HIV/AIDS pharmacotherapy. Efavirenz (EFV; aqueous solubility 4 microg/ml, bioavailability 40-45%) is a first-line agent in the pediatric therapeutic cocktail. The liquid formulation of EFV is not available worldwide, preventing appropriate dose adjustment and more convenient administration. The bioavailability of liquid EFV is lower than that of the solid formulation. Improving the bioavailability of the drug would reduce the cost of treatment and enable less affluent patients to access this drug. AIM: To encapsulate EFV in polymeric micelles to improve the aqueous solubility and the the oral bioavailability of the drug. METHODS: EFV was incorporated into the core of linear and branched poly(ethylene oxide) poly(propylene oxide) block copolymer micelles. The size and size distribution of the drug-loaded aggregates were characterized by dynamic light scattering and the morphology by transmission electron microscopy. The bioavailability of the EFV loaded micellar system (20 mg/ml) was assessed in male Wistar rats (40 mg/kg) and compared to that of a suspension prepared with the content of EFV capsules in 1.5% carboxymethylcellulose PBS solution (pH 5.0), and an EFV solution in a medium-chain triglyceride (Miglyol 812). RESULTS: This work demonstrates that the encapsulation of EFV, which is poorly water soluble, into polymeric micelles of different poly(ethylene oxide)-poly(propylene oxide) block copolymers significantly improves the oral bioavailability of the drug, and reduces the interindividual variability. CONCLUSION: This strategy appears a very promising one towards the development of a liquid aqueous EFV formulation for the improved pediatric HIV pharmacotherapy. PMID- 20025461 TI - Brain-targeted solid lipid nanoparticles containing riluzole: preparation, characterization and biodistribution. AB - AIM: Developments within nanomedicine have revealed a great potential for drug delivery to the brain. In this study nanoparticulate systems as drug carriers for riluzole, with sufficiently high loading capacity and small particle size, were prepared to a reach therapeutic drug level in the brain. MATERIALS & METHOD: Solid lipid nanoparticles containing riluzole have great potential as drug delivery systems for amyotrophic lateral sclerosis and were produced by using the warm oil-in-water microemulsion technique. The resulting systems obtained were approximately 88 nm in size and negatively charged. Drug-release profiles demonstrated that a drug release was dependent on medium pH. Biodistribution of riluzole blended into solid lipid nanoparticles was carried out after administration to rats and the results were compared with those obtained by riluzole aqueous dispersion administration. Rats were sacrificed at time intervals of 8, 16 and 30 h, and the riluzole concentration in the blood and organs such as the brain, liver, spleen, heart and kidney was determined. RESULTS: It was demonstrated that these solid lipid nanoparticles were able to successfully carry riluzole into the CNS. Moreover, a low drug biodistribution in organs such as the liver, spleen, heart, kidneys and lung was found when riluzole was administered as drug-loaded solid lipid nanoparticles. CONCLUSION: Riluzole loaded solid lipid nanoparticles showed colloidal size and high drug loading, a greater efficacy than free riluzole in rats, a higher capability to carry the drug into the brain and a lower indiscriminate biodistribution. PMID- 20025462 TI - Biosynthesis of antimicrobial silver nanoparticles by the endophytic fungus Aspergillus clavatus. AB - AIM: To induce the biosynthesis of silver nanoparticles (AgNPs) using Aspergillus clavatus and evaluate their antimicrobial potential. MATERIALS & METHODS: Aspergillus clavatus (AzS-275), an endophytic fungus isolated from sterilized stem tissues of Azadirachta indica A. Juss., was challenged with 1 mM AgNO(3) solution. The characterization of the AgNPs was carried out by x-ray diffraction spectrometry, transmission-electron microscopy and atomic force microscopy. RESULTS & DISCUSSION: The synthesized AgNPs were found to be extracellular, polydispersed spherical or hexagonal particles ranging from 10 to 25 nm in size. Antimicrobial activity was performed using a disc-diffusion method against Candida albicans, Pseudomonas fluorescens and Escherichia coli. The results showed an average minimum inhibitory concentration of 5.83 microg ml(-1) and minimum fungicidal concentration of 9.7 microg ml(-1) against C. albicans. CONCLUSIONS: AgNPs can be mycosynthesized extracellularly using A. clavatus as the fungal system, which is highly advantageous over chemical synthesis not only because it can be synthesized on a large scale, but because of the ease of downstream processing and its biomedical application in antimicrobial activity. PMID- 20025463 TI - Bone marrow-targeted liposomal carriers: a feasibility study in nonhuman primates. AB - BACKGROUND & AIMS: Recently, we described a novel surface-modified lipid vesicle formulation (liposome) that had very high targeting to bone marrow in normal rabbits. Because the bone marrow is the site of hematopoiesis, bone marrow targeted drug-delivery systems have many potential applications. In this study we investigated whether these bone marrow-targeted vesicles are also similarly effective for bone marrow targeting in rhesus monkeys, a primate animal model that is more relevant to humans. MATERIALS & METHODS: The preformed vesicles encapsulating 30 mM glutathione were labeled with technetium-99m ((99m)Tc) for scintigraphic imaging. The vesicles were 216 +/- 21 nm in diameter with a negative surface charge composed of DPPC, cholesterol, anionic amphiphile and poly(ethylene glycol)-DSPE (1:1:0.2:0.013 molar ratio). RESULTS: The whole-body images of rhesus monkeys receiving intravenous (99m)Tc vesicles revealed high uptake of the (99m)Tc vesicles in bone marrow. Based on image analysis, we estimated that approximately 70% of the injected dose of the (99m)Tc vesicles was taken up by the bone marrow. CONCLUSION: This finding increases the feasibility of using this bone marrow-specific drug-delivery system for clinical applications. PMID- 20025464 TI - DNA damage and p53-mediated growth arrest in human cells treated with platinum nanoparticles. AB - AIM: Platinum-based therapeutic agents are widely used in medicine. Thus, a thorough understanding of their mechanism of action in cells is warranted. This study investigates the uptake and bioactivity (e.g., cytotoxicity, genotoxicity and protein expression) of platinum nanoparticles (Pt-NPs, approximately 5-8 nm in size) in human cells. MATERIALS & METHODS: Pt-NPs capped with polyvinyl alcohol were synthesized, characterized and incubated with human cells. Uptake and the biological properties were evaluated through metabolic activity, genome integrity, cell cycle and protein expression. RESULTS: Pt-NPs entered the cells through diffusion, and localized inside the cytoplasm. Exposure to the Pt-NP increased DNA damage, accumulation of cells at the S-phase of the cell cycle and apoptosis. A significant number of cells recovered from the stress and formed colonies. Protein-expression levels uncovered upregulation of p53, phosphorylated p53, p21 and downregulation of proliferating cell nuclear antigen following Pt-NP treatment. Pro-caspase 3 and poly-ADP ribose polymerase and cyclin B levels were not altered in both the cell types after Pt-NP exposure. CONCLUSION: The results suggest p53 activation in Pt-NP-treated cells due to genotoxic stress, with subsequent activation of p21 leading to a proliferating cell nuclear antigen mediated growth arrest and apoptosis. This study recommends development of Pt-NP based anticancer agents by appropriate surface modifications to augment its innate anticancer activity. PMID- 20025465 TI - Time-varied magnetic field enhances transport of magnetic nanoparticles in viscous gel. AB - AIM: The potential of magnetic nanoparticles (MNPs) to deliver various forms of therapy has not been fully realized, in part due to difficulties in transporting the carriers through soft tissue to different target sites. The aim of this study was to demonstrate that transport of MNPs through a viscous gel can be controlled by a combined AC (time-varying) magnetic field and static field gradient. MATERIALS & METHODS: MNP velocity and transport efficiency were measured in a viscous gel at various settings of magnetic field and magnetite loadings. RESULTS: Combined application of an AC magnetic field with the static field gradient resulted in a nearly 30-fold increase in MNP transport efficiency in viscous gel for 30% (w/w) magnetite-loaded particles as compared with static field conditions. CONCLUSION: The 'oscillating' effect of an AC magnetic field greatly improves the ability to transport MNPs within soft media by decreasing the effective viscosity of the gel. PMID- 20025466 TI - Shape-coded silica nanotubes for multiplexed bioassay: rapid and reliable magnetic decoding protocols. AB - AIMS: The recent development of 1D barcode arrays has proved their capabilities to be applicable to highly multiplexed bioassays. This article introduces two magnetic decoding protocols for suspension arrays of shape-coded silica nanotubes to process multiplexed assays rapidly and easily, which will benefit the minimization and automation of the arrays. METHODS: In the first protocol, the magnetic nanocrystals are incorporated into the inner voids of barcoded silica nanotubes in order to give the nanotubes magnetic properties. The second protocol is performed by trapping the barcoded silica nanotubes onto streptavidin-modified magnetic beads. RESULTS: The rapid and easy decoding process was demonstrated by applying the above two protocols to multiplexed assays, resulting in high selectivity. Furthermore, the magnetic bead-trapped barcode nanotubes provided a great opportunity to exclude the use of dye molecules in multiplexed assays by using barcode nanotubes as signals. CONCLUSION: The rapid and easy manipulation of encoded carriers using magnetic properties could be used to develop promising suspension arrays for portable bioassays. PMID- 20025467 TI - Physicochemical properties affecting cellular uptake of carbon nanotubes. AB - Carbon nanotubes (CNTs) are widely used for biomedical applications as intracellular transporters of biomolecules owing to their ability to cross cell membranes. In this article, we survey the reported literature and results of our published work in an attempt to provide a rational view of the various CNT internalization mechanisms. Essentially three uptake mechanisms (phagocytosis, diffusion and endocytosis) have been reported in the literature. In addressing the subject of cellular internalization of CNTs, the unique physicochemical characteristics of CNTs that influence and drive the cell uptake pathway are considered. According to available evidence, the degree of dispersion, the formation of supramolecular complexes and the nanotube length are crucial factors in determining the exact mechanism of cellular uptake. In conclusion, phagocytosis appears to be the internalization pathway for CNT aggregates, bundles, cluster or single dispersed nanotubes 1 microm or more in length; endocytosis is the internalization mechanism for nanotubes forming supramolecular structures; and diffusion is the internalization mechanism for submicron CNTs that do not form supramolecular complexes. This information may be relevant to the rational design of CNT-based carriers for cell therapy. PMID- 20025468 TI - Nanotechnology for treatment of stroke and spinal cord injury. AB - The use of nanotechnology in cell therapy and tissue engineering offers promising future perspectives for brain and spinal cord injury treatment. Stem cells have been shown to selectively target injured brain and spinal cord tissue and improve functional recovery. To allow cell detection, superparamagnetic iron-oxide nanoparticles can be used to label transplanted cells. MRI is then a suitable method for the in vivo tracking of grafted cells in the host organism. CNS, and particularly spinal cord, injury is accompanied by tissue damage and the formation of physical and biochemical barriers that prevent axons from regenerating. One aspect of nanomedicine is the development of biologically compatible nanofiber scaffolds that mimic the structure of the extracellular matrix and can serve as a permissive bridge for axonal regeneration or as a drug delivery system. The incorporation of biologically active epitopes and/or the utilization of these scaffolds as stem cell carriers may further enhance their therapeutic efficacy. PMID- 20025469 TI - Surface activation and targeting strategies of superparamagnetic iron oxide nanoparticles in cancer-oriented diagnosis and therapy. AB - The advanced fabrication and surface engineering of superparamagnetic iron oxide nanoparticles (SPIONs) could offer excellent physiochemical features for noninvasive tumor imaging and drug delivery. The key issues of realization of maximized selective cancer targeting of SPIONs are minimization of uptake by macrophages, preferential binding to cancerous cells over neighboring normal cells, visualization of tumor cells prior to and after treatment and triggered drug release into target cells in a controlled fashion. In this article, we summarize the current status of fabrication of multifunctional SPION-based nanodevices specially designed for cancer-oriented diagnosis and therapy, with a focus on potential malignancy-targeting ligands' identification and development as nanocarriers. A number of examples of passive and active targeting strategies- lymphoangiogenesis markers, cellular metabolite receptors, extracellular matrix component receptors, neuropeptide receptors and receptor-mediated bypass of the blood-brain barrier--are described in detail. PMID- 20025470 TI - Regulation of nanomedicines in the EU: distilling lessons from the pediatric and the advanced therapy medicinal products approaches. AB - As the market for nanomedicines in the EU is growing, the development of regulatory guidance in this area assumes priority. Currently, the nanomedicine market is poised at a critical stage wherein clear regulatory guidance is imperative in providing for clarity and legal certainty to manufacturers of nanomedicine. The regulation of the pharmaceutical sector in the EU has witnessed several developments and innovations guided by the philosophy of single market and balancing the principle of ensuring high public health protection and safety of medicines. Both the pediatric and the advanced therapies medicinal products (ATMP) regimes offer important regulatory guidance that could be adopted for the regulation of nanomedicines in the EU. PMID- 20025473 TI - Utility of racemase and other immunomarkers in the detection of adenocarcinoma in prostatic tissue damaged by high intensity focused ultrasound therapy. AB - AIMS: High intensity focused ultrasound (HIFU) is an emerging alternative for the treatment of prostate adenocarcinoma. Alpha-methylacyl-CoA racemase (AMACR) has been shown to be a sensitive immunomarker for prostate cancer, however, there is no information available concerning its utility and that of other immunomarkers for the detection of malignancy after HIFU therapy. METHODS: AMACR expression was examined in 11 cases of prostatic carcinoma treated by HIFU, with histological evidence of residual carcinoma. In seven cases tumour was examined from thin core biopsies and in four cases from tissue fragments obtained by transurethral resection of prostate (TURP). In addition to AMACR, immunostaining was also undertaken for p63, cytokeratin 34betaE12, cytokeratin 5, cytokeratin 8-18, prostate specific alkaline phosphatase (PSAP), prostate specific antigen (PSA), chromogranin and CD56. RESULTS: In two of the cases foci of tumour were cut out in serial sections. AMACR was expressed in eight of nine evaluable cases (4/5 biopsies and 4/4 TURP specimens). Cytokeratin 8-18 and PSAP were positive in all cases, whereas PSA was positive in five of nine cases. Cytokeratin 34betaE12, cytokeratin 5, and p63 marked the basal layer in normal prostatic glands, but were negative in neoplastic glands. In four cases we found tumour cells with positive staining for CD56 and chromogranin. CONCLUSIONS: A panel with positive markers for AMACR, and negative markers for p63/cytokeratin 5/cytokeratin 34betaE12 confirms the neoplastic nature of the residual glands on biopsies or TURP fragments sampled after HIFU therapy. PMID- 20025471 TI - Programmable nano-bio-chips: multifunctional clinical tools for use at the point of-care. AB - A new generation of programmable diagnostic devices is needed to take advantage of information generated from the study of genomics, proteomics, metabolomics and glycomics. This report describes the 'programmable nano-bio-chip' with potential to bridge the significant scientific, technology and clinical gaps through the creation of a diagnostic platform to measure the molecules of life. This approach, with results at the point-of-care, possesses capabilities for measuring such diverse analyte classes as cells, proteins, DNA and small molecules in the same compact device. Applications such as disease diagnosis and prognosis for areas including cancer, heart disease and HIV are described. New diagnostic panels are inserted as 'plug and play' elements into the modular platform with universal assay operating systems and standard read out sequences. The nano-bio chip ensemble exhibits excellent analytical performance and cost-effectiveness with extensive validation versus standard reference methods (R(2) = 0.95-0.99). This report describes the construction and use of two major classes of nano-bio chip designs that serve as cellular and chemical processing units, and provides perspective on future growth in this newly emerging field of programmable nano bio-chip sensor systems. PMID- 20025474 TI - Diagnostic issues in second opinion consultations in prostate pathology. AB - AIM: The increase in early detection of prostate cancer in the Asian population has bolstered second opinion consultations in prostate pathology in this region. In this review, we aimed to identify the spectrum of lesions and queries submitted to a pathologist with uropathology interest at the Singapore General Hospital. METHOD: Request forms for second opinion prostate consultations from pathologists and clinicians were retrieved from central laboratory records and a specialist's correspondence files within 2004-2007. Histomorphological queries raised in the referrals, and comparison of original diagnosis and Gleason scoring with the review diagnoses were collated. RESULTS: Discordant diagnoses (183/323, 57%) were more common than concordant diagnoses (143/326, 44%) between original diagnosis and subspecialist review. The majority of discordances comprised initial undergrading of Gleason scores (132/183, 72%; p < 0.01) especially in needle core biopsies. Among the significantly altered pathological diagnoses, 24 cases (13%; p < 0.01) were changed from benign to malignant (n = 9, 5%; p < 0.01), and malignant to benign (n = 11, 6%; p < 0.01) or high grade prostatic intraepithelial neoplasia (n = 4, 2%; p < 0.01). Benign mimics of malignancy such as atypical adenomatous hyperplasia and atrophy, and small foci of adenocarcinoma, were some examples of morphological pitfalls. CONCLUSION: Prostate biopsy review is important because of management and prognostic implications that vary among Gleason scores, and predictive parameters that are detailed in prostate pathology reports. PMID- 20025475 TI - Immunolocalisation of fibrin in coronary atherosclerosis: implications for necrotic core development. AB - BACKGROUND: Intraplaque haemorrhage has been shown to be important in necrotic core enlargement. Immunolocalisation of fibrin within progressive stages of plaque progression has not been extensively studied. METHODS: Histological sections (n = 74) of human coronary arteries were stained immunohistochemically for fibrin II, red blood cell antigen (glycophorin A), and CD31. Plaques were chosen to represent a range of lesions [6 adaptive intimal thickening, AIT (AHA grade I); 4 intimal xanthomas (AHA grade II), 19 pathologic intimal thickening, PIT (AHA grade III, or pre-atheroma); 34 fibroatheromas, FA (AHA grade IV and V); and 11 thin cap fibroatheromas (TCFA, AHA grade IV)]. RESULTS: Fibrin was generally absent in the intima of AIT and PIT, with moderate staining in cores of early FA (2.6 +/- 0.3). All late FA and TCFA demonstrated intracore fibrin, with mean scores of 2.9 +/- 0.3 and 3.0 +/- 0.3, respectively. Intimal vasa vasorum counts increased with intimal fibrin score (p < 0.0001); in 68% of cores with fibrin staining, there was minimal or no evidence of red cell breakdown. CONCLUSIONS: Fibrin in necrotic cores is present proportional to intraplaque vasa vasorum and before red cells, suggesting leakage of vessels before frank intraplaque haemorrhage. Fibrin may play a role in the bridge between pre atheroma and atheroma. PMID- 20025476 TI - A review of histological outcomes from peri-menopausal and post-menopausal women with a cytological report of possible high grade abnormality: an alternative management strategy for these women. AB - AIM: To investigate the histological outcomes of women with cytological reports of possible high grade abnormality and to determine if patient age may be a clinically significant factor in determining the presence of high grade disease for this cytological category. METHODS: Using annual quality reports supplied by the Queensland Pap Test Register, the histological outcomes for 2054 women with possible high grade cytological reports over a 4 year period were investigated to determine the clinical significance of this category in older aged women. Women in the total study group were aged between 16 and 84 years and outcomes were divided into women aged under 40 years and 40 years and over as well as those aged under 50 years and 50 years and over. Cytomorphological features were compared to determine if there were any significant differences between the outcome groups. RESULTS: The histological confirmation rate for high grade disease for all women was 53.1%. The incidence of histologically confirmed high grade disease was significantly higher in women aged under 40 years (59.7%) compared to women aged 40 years and over (40.3%) (p = 0.001). This was also true for women aged under 50 years compared to those aged 50 years and over (55.8% versus 37.5%) (p = 0.001). When the smears from these women were reviewed, a predominance of mature intermediate and superficial cells was evident in the background of the majority of slides in all histological outcome categories. Many of the smears in the negative outcome group showed reactive, reparative or inflammatory changes. CONCLUSIONS: The currently mandated Australian management guideline for women with a cytology report of possible high grade disease is referral for colposcopic and histological examination. Since the incidence of high risk human papillomavirus (HPV) decreases with age, a less invasive strategy may be to test women in these older age groups for high risk HPV before proceeding to colposcopy. PMID- 20025477 TI - A class discovery and class prediction approach to histopathological classification of mammographic screen detected columnar cell lesions of the breast. AB - AIMS: Columnar cell lesions (CCLs) of the breast have been increasingly recognised in biopsies taken to investigate mammographic screen detected microcalcification. The aim of this study was to identify distinct CCL subtypes by systematic analysis of histopathology. METHODS: Hierarchical cluster analysis was performed based on the profile of histopathological features in 102 screen detected CCLs. Features assessed included nuclear morphology, acinar dilatation, epithelial cell hyperplasia, cell crowding, apical snout formation and intraluminal secretion. The stability of this classification was tested in an independent cohort of 32 cases. RESULTS: The histopathology of screen detected CCLs was extremely variable. Hierarchical cluster analysis identified two subclasses: Class 1 (34/102, 33%) characterised by absence of nuclear atypia and less pronounced hyperplasia; and Class 2 (68/102, 67%) that were generally more atypical. Ki-67 scores were significantly lower for Class 1 CCLs (p < 0.001). In the independent cohort of 32 cases, Class 1 cases were clearly distinguished from Class 2, indicating that these were stable phenotypes amongst screen detected CCLs. CONCLUSIONS: The histopathological features of CCLs diagnosed at screening are extremely heterogeneous. Using a systematic approach, we have devised a broad classification system that delineates a category of less atypical CCLs that could form a basis for future studies. PMID- 20025478 TI - Prognostic significance of apoptotic cell death in bladder cancer: a tissue microarray study on 179 urothelial carcinomas from cystectomy specimens. AB - AIMS: To assess the prognostic significance of apoptosis related markers in bladder cancer. METHODS: A tissue microarray containing 179 bladder carcinomas from cystectomy specimens was analysed immunohistochemically for active caspase 3, single-stranded DNA (ssDNA), p53, Bcl-2, Bax, and COX-2, in correlation to clinicopathological factors. RESULTS: Active caspase-3, ssDNA, p53, Bax and COX-2 were more frequently observed among high grade and higher stage (> or =T2) carcinomas compared with low grade and lower stage (T1) tumours. On the contrary, Bcl-2 was more frequently detected in T1 than in > or =T2 carcinomas. Active caspase-3 correlated with a better survival of the patients. CONCLUSIONS: The decreased detection of active caspase-3 and ssDNA and the increased presence of Bcl-2 in T1 carcinomas suggest that alterations in interrelated apoptosis markers may play an important role in the progression of urothelial carcinoma from a superficially infiltrating to a muscle invading tumour and would help to better characterise a subpopulation of T1 carcinomas that could profit from early cystectomy or more aggressive adjuvant chemotherapy. Active caspase-3 might be an important prognostic factor in bladder cancer. PMID- 20025479 TI - Annexin-I overexpression is associated with tumour progression and independently predicts inferior disease-specific and metastasis-free survival in urinary bladder urothelial carcinoma. AB - AIMS: In our previous studies, comparative proteomics and immunohistochemistry (IHC) demonstrated that annexin-I (ANXA1) is up-regulated in high grade urinary bladder urothelial carcinoma (UBUC) as compared to non-high grade carcinomas. However, the small sample size prohibited further correlation of ANXA1 expression to tumour progression. Therefore, in the present study, 81 primary localised UBUC specimens of various grades and primary tumour (pT) status were examined for ANXA1 expression to further confirm the proteomics data and to clarify the relevance of ANXA1 expression level to the prognosis of UBUC. METHODS: IHC was implemented to investigate ANXA1 protein expression in 81 primary localised UBUC specimens. The association of ANXA1 expression with tumour progression and prognosis was analysed. RESULTS: Our data demonstrated that the ANXA1 expression level was strongly associated with an escalated pT status (p < 0.001) and a higher histological grade (p < 0.001), suggesting that ANXA1 might be related to tumour progression. Moreover, at the univariate level, ANXA1 overexpression, along with higher pT status and histological grade, significantly predicted disease-specific survival (DSS) and metastasis-free survival (MFS). More importantly, multivariate analyses revealed that the association of ANXA1 overexpression and prognosis remained significant for both DSS and MFS. CONCLUSION: The above results reinforced the comparative proteomics results and confirmed the prognostic role of ANXA1 in UBUC. PMID- 20025480 TI - Expression of CARD6, an NF-kappaB activator, in gastric, colorectal and oesophageal cancers. AB - AIMS: Activation of nuclear factor-kappa B (NF-kappaB) signalling is considered a crucial mechanism in the development of cancers. Caspase-associated recruitment domain 6 (CARD6) is a protein that activates NF-kappaB signalling evoked by RIP1, RIP2, Bcl-10 and MEKK. In this study, we analysed tissue expression of CARD6 protein in oesophageal squamous cell carcinoma (ESCC), gastric adenocarcinomas (GC) and colorectal adenocarcinomas (CRC). METHODS: We analysed the expression of CARD6 protein in 58 ESCC, 100 GC and 103 CRC patients' tissues by immunohistochemistry using a tissue microarray (TMA) approach. RESULTS: We found CARD6 immunostaining in cancer cells of ESCC (41/58; 70.7%), GC (45/100; 45.0%) and CRC (81/103; 78.6%). In the GC, intestinal-type GC (77.8%) showed higher expression of CARD6 than diffuse-type GC (20.0%) and mixed-type GC (50.0%). By contrast, corresponding normal epithelial cells of oesophagus (0%), stomach (8.0%) and colon (5.0%) displayed lower frequencies of CARD6 immunostaining. The CARD6 immunostaining was observed in nucleus/cytoplasm (ESCC) or cytoplasm (GC and CRC). The CARD6 expression was evident from an early TNM stage (stage I). CONCLUSION: The increased expression of CARD6 in ESCC, GC and CRC tissues compared to their corresponding normal cells suggested that neoexpression of CARD6 might be related to activation of NF-kappaB pathway in the cancers and might play a role in the development of most types of gastrointestinal cancers. PMID- 20025481 TI - WT-1 expression in gastrointestinal stromal tumours. AB - AIMS: Wilms' tumour-1 (WT-1) encodes a transcription factor originally identified as a tumour suppressor gene, whose mutations are responsible for tumorigenesis of Wilms' tumour. The overexpression of WT-1 has been demonstrated in variable tumours. In this study, WT-1 immunoreactivity was explored in 97 gastrointestinal stromal tumours (GISTs). METHODS: The expression of WT-1 was compared with other immunohistochemical markers of GIST and the association with clinicopathological features was also evaluated. For comparison, six melanomas and 71 soft tissue tumours were used. RESULTS: All 97 GISTs were positive for WT-1 and the staining intensity was strong in 59 (60.8%), moderate in 28 (28.9%) and weak in 10 cases (10.3%). Cytoplasmic staining with moderate to strong intensity was more frequent in GISTs positive for platelet-derived growth factor receptor-alpha (PDGFRA) (p = 0.04). However, WT-1 immunoreactivity was not related to the clinicopathological variables, including tumour location, histological subtype, risk of progression and recurrence status. Leiomyosarcomas, schwannomas, malignant peripheral nerve sheath tumours (MPNSTs) and melanomas showed cytoplasmic staining, whereas leiomyomas and mesenteric fibromatoses were totally negative for WT-1. CONCLUSIONS: Although the biological roles of WT-1 in GIST are still unknown, our findings might provide a rationale for immunotherapy targeting WT-1 and a therapeutic solution to the challenge of imatinib-resistant GISTs. PMID- 20025482 TI - Cell cycle regulators, APC/beta-catenin, NF-kappaB and Epstein-Barr virus in gastric carcinomas. AB - AIMS: To evaluate the clinicopathological value of cell cycle regulators, the Wnt pathway, the NF-betaB pathway and Epstein-Barr virus (EBV) and to assess their relationships in gastric carcinoma. METHODS: We investigated cell cycle regulators (p53, p21, Rb), APC, beta-catenin and NF-kappaB using immunohistochemistry and EBV using in situ hybridisation for EBV encoded small RNAs in 117 cases of gastric carcinoma. RESULTS: p53 overexpression was more frequently observed in advanced gastric carcinoma and lymph node metastasis than in early carcinoma or in the absence of metastasis (p < 0.05). p21 loss was positively correlated with APC loss, but inversely correlated with beta-catenin nuclear accumulation and NF-kappaB positivity (p < 0.05). EBV positive gastric carcinomas were located in the upper third of the stomach, and more were of the diffuse or mixed types than the EBV negative group (p < 0.05). EBV infection was positively correlated with p21 loss and APC loss and inversely correlated with beta-catenin alteration (p < 0.05). In multivariate analysis, patient age, TNM stage and p53 were independent prognostic factors for gastric carcinoma. CONCLUSIONS: p53 status is a prognostic marker for gastric carcinoma. p21, APC, beta-catenin and NF-kappaB may be functionally interrelated in gastric carcinogenesis. Loss of p21 and APC may be involved in the carcinogenesis of EBV positive gastric carcinomas. PMID- 20025484 TI - Mutational analysis of CASP10 gene in colon, breast, lung and hepatocellular carcinomas. AB - AIMS: Evasion of apoptosis is a feature of cancer cells. As a mechanism of apoptosis inactivation in cancer cells, somatic mutations of pro-apoptotic genes have been reported in many cancers. Caspase-10 is an initiation-phase caspase, and somatic mutation of CASP10 that encodes caspase-10 has been found in non Hodgkin's lymphoma and gastric carcinoma. METHODS: The aim of this study was to explore whether CASP10 gene is somatically mutated in colon, breast, lung, and hepatocellular carcinomas. We analysed the entire coding region and all splice sites of CASP10 in 47 colon, 47 breast, 47 lung, and 47 hepatocellular carcinomas by a single-strand conformation polymorphism (SSCP) assay. RESULTS: We found two CASP10 mutations in the colon cancers (2/47; 4.3%), but none in breast, lung or hepatocellular carcinomas. One mutation [c.41A > C (p.Lys14Thr)] was a missense mutation, while the other was a substitution mutation in a splice site (c.684 + 4G > A). The colon cancer with the CASP10 missense mutation harboured additional CASP gene mutations (CASP3, 7 and 8). CONCLUSION: Our data indicate that somatic mutation of CASP10 is rare in colon, breast, lung, and hepatocellular carcinomas. However, the data also suggest that CASP10 mutation might contribute to the pathogenesis of some colon carcinomas together with other CASP gene mutations. PMID- 20025483 TI - Olive leaf extract prevents spontaneous occurrence of non-alcoholic steatohepatitis in SHR/NDmcr-cp rats. AB - AIMS: Oxidative stress may play an important role in the pathogenesis of non alcoholic steatohepatitis (NASH). Oleuropein, the active constituent of olive leaf, possesses anti-oxidant, hypoglycaemic, and hypolipidaemic activities. We aimed to investigate the preventive effects of olive leaf extract on hepatic fat accumulation in a rat model of NASH. METHODS: Spontaneously hypertensive/NIH corpulent rats were fed a diet of AIN-93G with or without olive leaf extract (500, 1000, 2000 mg/kg diet, and control; 5 rats each) for 23 weeks. Serological and histopathological findings, anti-oxidative activity, and the alteration of fatty acid synthesis in the liver were evaluated. RESULTS: Histopathologically, a diet of AIN-93G containing more than 1000 mg/kg olive leaf extract had a preventive effect for the occurrence of NASH. Thioredoxin-1 expression in the liver was more evident in rats fed this diet, and 4-hydroxynonenal expression in the liver was less evident in these rats. There were no significant differences in the activities of hepatic carnitine palmitoyltransferase, fatty acid synthase, malic enzyme, and phosphatidic acid phosphohydrolase among the groups. CONCLUSIONS: Our data suggest that olive leaf extract may help prevent NASH, presumably through its anti-oxidative activity. PMID- 20025485 TI - Citraconic anhydride: a new antigen retrieval solution. AB - BACKGROUND: The introduction of heat-induced antigen retrieval has been a major milestone in diagnostic immunohistochemistry, enabling the application of many antibodies to fixed paraffin-embedded tissue sections. A number of important variables affect the preservation of tissue antigens, among which are analytical variables including the antigen retrieval methodology. Temperature of retrieval, duration of heating, source of heat, pH and nature of retrieval solution are among more important variables pivotal to results. Citrate buffer at pH 6.0 has become widely embraced as the universal fixative but some antibodies remain capricious and yield poor staining results. METHODS: This study examines the recent suggestion that citraconic anhydride may be a suitable universal retrieval reagent. Immunostaining of 65 commonly employed antibodies following microwave antigen retrieval in 0.05% citraconic anhydride for 10 minutes at 98 degrees C was compared with consecutive tissues sections subjected to antigen retrieval in citrate buffer at pH 6.0 at the same duration and temperature. RESULTS: Thirty five of the 65 antibodies examined yielded more intense staining following antigen retrieval in citraconic anhydride, including some capricious antibodies such as MyoD1, myogenin, perforin, TIA-1, Tdt, RET and MiTF, confirming the efficacy of this retrieval solution. CONCLUSION: It is recommended that consideration be given to 0.05% citraconic anhydride as an antigen retrieval solution, particularly for antibodies that fail to work or stain weakly in fixed paraffin-embedded tissue sections. PMID- 20025486 TI - IgE-type multiple myeloma with the late development of IgA2 kappa and plasma cell leukaemia. PMID- 20025487 TI - Gelatinous transformation of the bone marrow as a late morphological change in imatinib mesylate treated chronic myeloid leukaemia. PMID- 20025488 TI - Aggressive systemic mastocytosis with Charcot-Leyden crystals-associated crystal storing histiocytosis in bone marrow. PMID- 20025489 TI - Leiomyosarcoma of the bone: a case report of a rare tumour and problems involved in diagnosis. PMID- 20025490 TI - A novel EXT1 gene mutation causing hereditary multiple exostoses in a Chinese pedigree. PMID- 20025491 TI - Mutational analysis of UBR5 gene encoding an E3 ubiquitin ligase in common human cancers. PMID- 20025492 TI - Genotype-confirmed alanine aminotransferase deficiency in a Chinese patient with acute liver failure: a potential diagnostic pitfall. PMID- 20025493 TI - Endometriosis with myxoid change mimicking pseudomyxoma peritonei. PMID- 20025494 TI - Hybrid endometrial carcinoma with short non-villous papillae and serous papillary carcinoma. PMID- 20025495 TI - Challenging diagnosis: florid epithelial hyperplasia versus adenocarcinoma of the fallopian tube. PMID- 20025496 TI - Primary small cell carcinoma of the ureter: a case report involving immunohistochemical and molecular genetic analyses of KIT and PDGFRA genes. PMID- 20025497 TI - Medulloblastoma arising from intracranial immature teratoma. PMID- 20025499 TI - Generation of novel anti-BTLA monoclonal antibodies for in vivo use and their functional testing at near-physiological conditions. AB - We have generated and characterized a panel of monoclonal antibodies recognizing B and T lymphocyte attenuator (BTLA), a transmembrane protein expressed on essentially all lymphoid cells. One of the monoclonal antibodies (MAbs) detects for the first time all BTLA protein variants described for various mouse strains with high sensitivity, both in flow cytometry and immunohistology. Further tests have determined that this MAb recognizes a BTLA epitope independent of the HVEM binding site. Moreover, we identified a number of antibodies capable of efficiently blocking the interaction of BTLA with its ligand herpes virus entry mediator (HVEM). A series of experiments was performed with these MAbs at near physiological conditions to assess their blocking potential in vivo. These tests, performed with whole MAbs and also their F(ab)(2) formats, revealed that measurements of binding at 37 degrees C to primary cells expressing the target protein on the cell surface offer superior information on their blocking capacity. The generated BTLA-specific MAb will be used for in vivo studies to further elucidate the biological role of BTLA-HVEM interaction and function in vivo. PMID- 20025498 TI - Detection of amino-terminal extracellular domain of somatostatin receptor 2 by specific monoclonal antibodies and quantification of receptor density in medulloblastoma. AB - Somatostatin receptor 2 (SSTR2) is expressed by most medulloblastomas (MEDs). We isolated monoclonal antibodies (MAbs) to the 12-mer (33)QTEPYYDLTSNA(44), which resides in the extracellular domain of the SSTR2 amino terminus, screened the peptide-bound MAbs by fluorescence microassay on D341 and D283 MED cells, and demonstrated homogeneous cell-surface binding, indicating that all cells expressed cell surface-detectable epitopes. Five radiolabeled MAbs were tested for immunoreactive fraction (IRF), affinity (KA) (Scatchard analysis vs. D341 MED cells), and internalization by MED cells. One IgG(3) MAb exhibited a 50-100% IRF, but low KA. Four IgG(2a) MAbs had 46-94% IRFs and modest KAs versus intact cells (0.21-1.2 x 10(8) M(-1)). Following binding of radiolabeled MAbs to D341 MED at 4 degrees C, no significant internalization was observed, which is consistent with results obtained in the absence of ligand. However, all MAbs exhibited long-term association with the cells; binding at 37 degrees C after 2 h was 65-66%, and after 24 h, 52-64%. In tests with MAbs C10 and H5, the number of cell surface receptors per cell, estimated by Scatchard and quantitative FACS analyses, was 3.9 x 10(4) for the "glial" phenotype DAOY MED cell line and 0.6-8.8 x 10(5) for four neuronal phenotype MED cell lines. Our results indicate a potential immunotherapeutic application for these MAbs. PMID- 20025500 TI - A monoclonal antibody for detection of folylpolyglutamate synthetase in paraffin embedded tissues. AB - Folate biochemical pathway enzymes such as folylpolyglutamate synthetase (FPGS) are key elements in the folate pathway. The role of FPGS is to add glutamate residues to folates and antifolates, trapping them in the cell and increasing their affinity for subsequent enzymatic reactions. FPGS may also be an indicator of response to both clinically established and novel antifolate drugs such as pemetrexed; knowledge of their level of expression in tumors may enable their optimal use by identifying potentially responsive subgroups of patients. In spite of its key role in both nucleotide biosynthesis and possible role as a determinant of response in chemotherapy, monoclonal antibodies to FPGS suitable for immunohistochemical analysis of formalin fixed and paraffin embedded biopsy samples, or that can be used for Western blot analysis, are not commercially available. The aim of this study was to generate a monoclonal antibody that could be used to detect specific expression of FPGS in paraffin embedded tissues. A 228 amino acid region of the FPGS sequence was expressed as a recombinant fusion protein and used as an antigen to generate monoclonal antibodies. ELISA and Western blot studies identified specific reactivity of the NN3.2 antibody to the recombinant protein and a single 60 kDa protein in whole cell lysates from cell lines known to express FPGS. Immunohistochemical analysis of FPGS using hybridoma clone NN3.2 in a panel of normal tissues demonstrated wide expression including strong immunoreactivity in the brush border and crypts of colon, liver hepatocytes, and lymphoid cells. Analysis of a panel of malignant and benign tissues demonstrated wide expression with variable intensities of staining and patterns of cytoplasmic reactivity. Stronger staining was observed in malignant tissue compared with that of normal adjacent tissue, particularly in ovarian and colon adenocarcinoma cases. Our results show that clone NN3.2 is a sensitive tool for detection of FPGS in paraffin-embedded tissues. PMID- 20025501 TI - A new monoclonal antibody recognizing a linear determinant on the HLA-DRalpha chain N-terminus. AB - We report the generation of a monoclonal antibody (MAb) that reacts to the N terminus of the denatured HLA-DRalpha chain. The 1C4.6 MAb was raised against a peptide corresponding to amino acid residues 10 to 32 of a highly conserved region within the alpha1 domain of HLA-DR. This region partially overlaps with the epitope recognized by the conformationally dependent L243 MAb. In Western blot analysis, MAb 1C4.6 reacted with denatured HLA-DRalpha chains, but failed to bind the HLA-DRbeta chain expressed individually by transfectant cells, confirming that it recognizes an epitope on the alpha-chain of HLA-DR. In addition, this antibody was found to be isotype specific to HLA-DRalpha, as it did not cross-react to HLA class II proteins HLA-DP and-HLA-DQ. The 1C4.6 MAb is a valuable addition to existing reagents used to probe the structure and function of MHC class II molecules. This anti-HLA-DRalpha1 domain MAb may prove valuable for studies of HLA class II heterodimer assembly, structure, and function, as well as for studies into the release of soluble MHC class II. PMID- 20025502 TI - Monoclonal antibodies produced against lipopolysaccharide from fimA Type II Porphyromonas gingivalis. AB - An important periodontal pathogen, Porphyromans gingivalis strains are classified into six genotypes (types I-V and Ib), based on the genotype of the fimbriae A (fimA). Among the genotypes, fimA type II strains are thought to be most strongly related to advanced periodontitis. To develop passive immunotherapy, over 300 hybridoma clones were constructed through immunization of cell extracts of fimA type II strain P. gingivalis TDC60 using hybridoma technology. Among these clones, 15 MAbs recognized TDC60 lipopolysaccharide (LPS) with an individual ladder-like structure by Western blot analysis. Further Western blotting of the 15 MAbs against LPS from TDC60, FDC381 (fimA type I), and W83 (fimA type IV) of P. gingivalis and Escherichia coli was carried out. None of these MAbs recognized E. coli LPS, and divided into at least three different Western blot patterns. To confirm the specificity to LPS, three clones were selected and competition assays were carried out using TDC60 LPS. All three MAbs reduced the reactivity against TDC60 LPS after absorption of the LPS in a dose-dependent manner. These findings suggest that MAbs recognizing different epitopes of P. gingivalis LPS were successfully constructed, and these MAbs may be useful in neutralizing P. gingivalis infection. PMID- 20025503 TI - Generation of a monoclonal antibody specifically reacting with undifferentiated liver progenitor cells. AB - An adult rat liver progenitor cell line Lig-8 was established. In the induction by sodium butyrate, these cells were shown to be able to differentiate into both hepatocytes and bile duct cells expressing albumin and cytokeratin-19, the markers of respective cell types. In order to generate Lig-8 specific antibody for further studies, we produced a monoclonal antibody using the whole Lig-8 cells as immunogen. The yielded monoclonal antibody, named Ligab, belongs to IgG subclass G1 and kappa light chain. It specifically stained on Lig-8 cells in the cytoplasm but not on a rat hepatoma cell line H4IIE. Its immunoreaction against Lig-8 cell lysate on dot blots diminished as the concentration of sodium dodecyl sulfate (SDS) in the lysate increased to 2%, a level in the sample buffer of standard SDS-polyacrylamide gel electrophoresis (PAGE). Not surprisingly, Ligab failed to detect its reacting antigen in Lig-8 cell lysate by standard SDS-PAGE based immunoblotting. It could detect this antigen only by native PAGE-based immunoblotting. These characteristics suggested that the antigenic epitope for Ligab is likely a molecular structure instead of a peptide sequence. More interestingly, expression of Ligab-reacting antigen in Lig-8 cells declined as the cells were induced to differentiate by sodium butyrate. This antigen is very likely a differentiation-related marker for these cells, and this monoclonal antibody may help study the molecular mechanisms of liver progenitor cell differentiation. PMID- 20025504 TI - An anti-DLK1 monoclonal antibody produced using ELISA and hybridoma techniques. AB - DLK1 is a newly identified prognostic factor associated with liver cancer survival. To prepare specific monoclonal antibody (MAb) against DLK1, cDNA of DLK1 was cloned by RT-PCR and inserted into prokaryotic expression vector pGEX 4T1, respectively. The fusion proteins were expressed in Escherichia coli. Monoclonal antibody against DLK1 was obtained with hybridoma technique and specific ELISA screening. Western blotting and immunohistochemistry assays showed that MAb 6D6 had specific binding ability with DLK1 protein in eukaryotic cells and cancer tissues. This MAb will be a helpful tool for the detection of DLK1 protein in the tissues and serum of liver cancer and other cancer patients. PMID- 20025505 TI - Generation of a rat monoclonal antibody specific for glyoxalase I. AB - Glyoxalase I (GLO1) is a key enzyme that plays a role in the detoxification of methylglyoxal (MG), a toxic cellular metabolite produced during glycolysis. The present study reports on the preparation and properties of a monoclonal antibody (MAb) directed against mouse GLO1. The antibody was produced by hybridization of mouse myeloma cells with lymph node cells from an immunized rat. The MAb 6F10 specifically recognized GLO1, as evidenced by immunoblotting using a variety of extracts from cultured cells. In immunostaining using MAb 6F10, a diffuse cytoplasmic and nuclear staining pattern was observed. The MAb 6F10 promises to be useful in immunoblotting and immunostaining experiments in various cells and tissues to determine the expression levels of GLO1, as well as to further analyze the biological function of this protein. PMID- 20025506 TI - Generation of a rat monoclonal antibody specific for Pax7. AB - Pax7 is a nuclear localization protein, well known as a member of the paired box family. It is expressed at a very early stage of muscle differentiation and is also found in muscle satellite cells that are recognized as muscle stem cells. Pax7 is also recognized as a tumor cell marker since it is greatly expressed in various types of tumor cells. Pax7 has homology among other paired family members and is not easy to distinguish one from the others. In this study, we report on the establishment of monoclonal antibodies (MAb) against Pax7 using a rat medial iliac lymph node method. The quality of the antibody was examined by immunoblotting analysis. It was confirmed that the antibody can specifically recognize the Pax7 protein. It was also revealed that the MAb antibody successfully recognizes the nuclear localized Pax7 protein in Ewing's sarcoma cells by immunocytochemistry. The antibody can clearly show the regions of euchromatin and heterochromatin where hoechst is positive. PMID- 20025507 TI - Generation of a rat monoclonal antibody specific for Brm. AB - Brm is a subunit of the SWI/SNF complex that has a ATPase activity. It is well known that the complex plays a major role in cell processes, such as proliferation, differentiation, and DNA repair of cells. Here we report the production of monoclonal antibody (1H7A10) against Brm by rat medial iliac lymph node method. Immunoblot analysis with the antibody revealed the specific recognition of Brm and increase of Brm protein level in skeletal muscle differentiation. Immunocytochemistry analysis shows nuclear localization in myoblast C2C12 and involvement of transcription in the late stages of differentiation. PMID- 20025508 TI - Generation and characterization of novel monoclonal antibodies recognizing UBXD1. AB - UBXD1 is a recently identified adaptor for p97, a highly abundant and conserved member of the AAA family of ATPase that plays pivotal roles in a multitude of cellular processes involving the ubiquitin-proteasome pathway. Very little is known about the biochemical, cellular, and molecular functions of UBXD1. Here we report the generation of two mouse monoclonal antibodies, 5C3-1 and 2F8-24, that recognize UBXD1 using Western blotting, immunoprecipitation, and immunofluorescence. PMID- 20025509 TI - Production of a rat monoclonal antibody against Brg1. AB - Brm-related gene-1 (Brg1) is a catalytic subunit of the SWI/SNF chromatin remodeling enzyme complex that has ATPase activity. This complex facilitates chromatin remodeling for gene expression by utilizing energy for ATP hydrolysis. It is well known that the SWI/SNF chromatin remodeling enzyme complex is essential for cell differentiation, cell cycle regulation, and embryogenesis. Here we report the establishment of a hybridoma cell line for producing an antibody against Brg1 subunit by the rat medial iliac lymph node method. Immunoblot analysis showed that our antibody can specifically recognize Brg1. It was revealed by immunocytochemistry that Brg1 is located in euchromatin of C2C12 myoblast nuclei. These data suggested this antibody is useful for analyzing molecular function of Brg1 protein in cells. PMID- 20025513 TI - Using the Internet to provide care for persons living with HIV. AB - There are no published reports on ways in which caregivers use the Internet to support people living with HIV/AIDS (PLWHA). Five hundred caregivers were recruited in a 5-week period to complete an online survey of demographic characteristics, Internet use, online health-seeking self-efficacy, and ways they used the Internet to support PLWHA. Caregivers were on average 39 years old, white, heterosexual, highly educated, and Internet-savvy. Most provided informal care only (e.g., as a friend; 78%), with the remainder divided among those who provided care exclusively as part of their job (11%) or in both informally and professionally (11%). Most (72%) respondents visited a general medical website for HIV information, and 44% shared information from the Internet with PLWHA. Compared to informal caregivers, caregivers whose roles were both informal and professional had greater odds of recently sharing information from the Internet with PLWHA (odds ratio [OR] = 2.03) and ever printing off information from a website to give to PLWHA (odds ratio [OR] = 3.87). Professional caregivers had higher odds of ever printing off information from a website to give to PLWHA (OR = 1.87), but lower odds of sending an e-mail with a website link (OR = 0.32) than informal caregivers. These findings suggest that websites providing HIV-related resources should consider the various ways in which caregivers use their content, and how utilization differs by role. More research is needed to understand how people providing care for PLWHA share information and support each other and the impact that doing so has on caregiver burden and treatment outcomes for PLWHA. PMID- 20025514 TI - Symptoms of psychological distress: a comparison of rural and urban individuals enrolled in HIV-related mental health care. AB - Over the past decade, the number of individuals who have been diagnosed with HIV in nonmetropolitan areas (population of less than 50,000 individuals) has increased; however, the majority of the research has been conducted in metropolitan areas. Even less research has examined the levels of psychological distress among rural individual living with HIV. The purpose of this study was to explore the nature and range of psychological distress symptoms experienced by individuals living in rural areas who had self-enrolled into HIV-related mental health care and to compare their levels of distress to their urban counterparts accessing care at the same clinic. Data were collected from 95 individuals who self-enrolled in HIV-related mental health at either a rural (n = 47) or urban (n = 48) clinic. All participants completed the Brief Symptom Inventory (BSI), the instrument used in this study to assess symptoms of psychological distress. Rural participants had significantly higher mean scores on the hostility dimension of the BSI, F(1, 93) = 8.77, p = 0.004, than their urban counterparts. Furthermore, the rural participants had a greater proportion of individuals who had a T-score >or=63, a level indicative of a need for further psychological evaluation, for generalized anxiety, hostility, and psychoticism. The results indicated that rural individuals presented with higher levels of symptoms of psychological distress than their urban counterparts. These differences may be reflective of situational circumstances in rural areas where access to care, social isolation, and perceived stigma may delay screening for, and treatment of, psychological distress. PMID- 20025515 TI - Evaluation of a health setting-based stigma intervention in five African countries. AB - The study aim is to explore the results of an HIV stigma intervention in five African health care settings. A case study approach was used. The intervention consisted of bringing together a team of approximately 10 nurses and 10 people living with HIV or AIDS (PLHA) in each setting and facilitating a process in which they planned and implemented a stigma reduction intervention, involving both information giving and empowerment. Nurses (n = 134) completed a demographic questionnaire, the HIV/AIDS Stigma Instrument-Nurses (HASI-N), a self-efficacy scale, and a self-esteem scale, both before and after the intervention, and the team completed a similar set of instruments before and after the intervention, with the PLHA completing the HIV/AIDS Stigma Instrument for PLHA (HASI-P). The intervention as implemented in all five countries was inclusive, action-oriented, and well received. It led to understanding and mutual support between nurses and PLHA and created some momentum in all the settings for continued activity. PLHA involved in the intervention teams reported less stigma and increased self esteem. Nurses in the intervention teams and those in the settings reported no reduction in stigma or increases in self- esteem and self-efficacy, but their HIV testing behavior increased significantly. This pilot study indicates that the stigma experience of PLHA can be decreased, but that the stigma experiences of nurses are less easy to change. Further evaluation research with control groups and larger samples and measuring change over longer periods of time is indicated. PMID- 20025519 TI - Antifibrotic effect of hepatocyte growth factor-expressing mesenchymal stem cells in small-for-size liver transplant rats. AB - Ischemia-reperfusion and chronic injuries associated with small-for-size liver transplantation (SFSLT) impair the regeneration of liver graft and induce liver fibrosis. Mesenchymal stem cells (MSCs) can prevent the development of liver fibrosis, and hepatocyte growth factor (HGF) can also attenuate liver cirrhosis. Our previous studies have demonstrated that higher occurrence of liver fibrosis existed in rats post-SFSLT, and that implantation of HGF/MSCs, the human HGF (hHGF)-expressing MSCs, can improve liver regeneration, reduce mortality of rats, as well as have the potent antifibrotic effect in this SFSLT model. In the present study, we implanted HGF/MSCs into liver grafts via the portal vein and investigated their role in antifibrosis effect, using a 30% SFSLT rat model. Fibrosis indexes, including laminin (LN), hyaluronic acid (HA) levels in serum and hydroxyproline (Hyp) content in the liver grafts, the expression of transforming growth factor-beta1 (TGF-beta(1)), rat HGF (rHGF), alpha-smooth muscle actin (alpha-SMA) in hepatic stellate cells (HSCs), alanine aminotransferase (ALT), total bilirubin (BIL), and albumin (ALB) levels in serum, in rats in different treatment groups were assessed at different time points. We found that HGF/MSCs significantly inhibited the formation of liver fibrosis in rats undergoing SFSLT, while MSCs and HGF had synergistic effects in the process. The antifibrosis effect of HGF/MSCs may have contributed in modulating the activation and apoptosis of HSCs, elevating the rHGF expression level, and decreasing the TGF-beta(1) secretion of activated HSCs. These studies suggest that HGF/MSCs may be a novel therapeutic option for the treatment of liver fibrosis after SFSLT. PMID- 20025520 TI - Pluripotency and reprogramming: meeting report on the Fifth International Meeting of the Stem Cell Network North Rhine Westphalia. PMID- 20025521 TI - Dynamic epigenetic regulation of the Oct4 and Nanog regulatory regions during neural differentiation in rhesus nuclear transfer embryonic stem cells. AB - Oct4 and Nanog are crucial for maintaining pluripotency in embryonic stem (ES) cells and early-stage embryos. In the present study, the status of DNA methylation and of histone modifications in the regulatory regions of Oct4 and Nanog in rhesus nuclear transfer-derived ES (ntES) cells was compared with in vitro fertilized embryo-derived ES (IVFES) cell counterparts. Dynamic changes in DNA methylation during differentiation into neural lineage were also monitored and correlated with mRNA abundance and protein levels of both genes. In ntES cells Oct4 exhibited mono-allelic methylation along with relatively lower mRNA levels, and its transcription was seen predominantly from the unmethylated allele. In contrast, in IVFES cells Oct4 was hypomethylated on both alleles and had relatively higher transcript levels, suggesting incomplete reprogramming of DNA methylation on the Oct4 gene following somatic cell nuclear transfer. During neuronal differentiation, Oct4 underwent biallelic methylation and reduced amounts of Oct4 mRNA were detected in both types of ES cells. Analysis of Nanog regulatory regions revealed that both alleles were hypomethylated and similar levels of Nanog transcripts were expressed in ntES cells and IVFES cells. During neuronal differentiation both alleles were methylated and reduced amounts of Nanog mRNA were detected. Other epigenetic modifications including histone 3 lysine 4, 9, and 27 trimethylation (H3K4me3, H3K9me3, and H3K27me3) showed similar patterns around the regulatory regions of Oct4 and Nanog in both kinds of ES cells. During neural differentiation, dramatic enrichment of H3K27me3 and H3K9me3 (repressive marks) was observed on Oct4 and Nanog regulatory regions. Differentiation of ntES and IVFES cells correlated with the silencing of Oct4 and Nanog, reactivation of the neural marker genes Pax6, N-Oct3, and Olig2, and dynamic changes in histone modifications in the upstream regions of Pax6 and N Oct3. In short, although ES cells derived from somatic cell nuclear transfer showed a different epigenetic status in the Oct4 regulatory region than the IVF derived counterparts, based on the parameters tested, the neural differentiation potential of ntES and IVFES cells is equivalent. PMID- 20025522 TI - High-efficiency production of subculturable vascular endothelial cells from feeder-free human embryonic stem cells without cell-sorting technique. AB - We previously reported a feeder-free culture method for pure production of subculturable vascular endothelial cells (VECs) from cynomolgus monkey embryonic stem cells (cmESCs) without as using cell-sorting technique. By this method, canonical vascular endothelial (VE)-cadherin/platelet-endothelial cell adhesion molecule 1 (PECAM1)-positive VECs (c-VECs) and atypical VE-cadherin/PECAM1 negative VECs (a-VECs) were generated without a contamination by pericytes, lymphatic endothelial cells, or immature ES cells. More recently, we established a unique culture technique to maintain human ESCs (hESCs) under a feeder-free and recombinant cytokine-free condition. Combining these two systems, we have successfully generated pure VECs from two lines of hESCs, khES-1 and khES-3, under a completely feeder-free condition. Our method is very simple: spheres generated from hESCs by floating culture using differentiation media supplemented with vascular endothelial growth factor, bone morphogenetic protein 4, stem cell factor, FMS-related tyrosine kinase-3 ligand, and interleukin 3 (IL3) and IL6 were cultured on gelatin-coated plates. Cell passage was performed by an ordinary enzymatic treatment. The hESC-derived differentiated cells demosntrated cord forming activities and acetylated low-density lipoprotein-uptaking capacities. Moreover, they exclusively expressed von Willebrand factor and endothelial nitric oxide synthase. Flow cytometric analyses indicate that khES-3 generated both c VECs and a-VECs as in the case of cmESCs. By contrast, khES-1 produced only a VECs, which nonetheless demonstrated effective recruitment into neovascularity in vivo. Interestingly, a-VECs turned to express PECAM1 after transplantation into immunodeficient mice. The hESC-derived VECs were subculturable at least up to 10 passages without functional depression. Our method does not require a presorting processes to enrich progenitor fractions such as CD34-positive or kinase insert domain receptor (KDR)-positive cells, providing the most efficient and easiest technique for VEC production from hESCs. PMID- 20025523 TI - Enhanced generation of myeloid lineages in hematopoietic differentiation from embryonic stem cells by silencing transcriptional repressor Twist-2. AB - The self-renewal and multilineage differentiation of embryonic stem cells (ESC) is largely governed by transcription factors or repressors. Extensive efforts have focused on elucidating critical factors that control the differentiation of specific cell lineages, for instance, myeloid lineages in hematopoietic development. In this study, we found that Twist-2, a basic helix-loop-helix (bHLH) transcription factor, plays a critical role in inhibiting the differentiation of ESC. Murine ES cells, in which Twist-2 expression is silenced by lentivirally delivered shRNA, exhibit an enhanced formation of primary embryoid bodies (EB) and enhanced differentiation into mesodermally derived hematopoietic colonies. Furthermore, Twist-2 silenced (LV-siTwist-2) ESC display significantly increased generation of myeloid lineages (Gr-1(+) and F4/80(+) cells) during in vitro hematopoietic differentiation. Treatment with the Toll like receptor (TLR) 4 ligand synergistically stimulates the generation of primary EB formation as well as of hematopoietic progenitors differentiated from LV siTwist-2 ES cells. Thus, this study reveals the critical role of the transcriptional repressor Twist-2 in regulating the development of myeloid lineage in hematopoietic differentiation from ESC. This study also suggests a potential strategy for directional differentiation of ESC by inhibiting a transcriptional repressor. PMID- 20025524 TI - Identification of multipotent cytotrophoblast cells from human first trimester chorionic villi. AB - In this article we used immunohistochemistry and FACS analyses to show that cells expressing markers typical of human stem cells such as SSEA4, OCT-4, ALP, and CD117 are present within the cytotrophoblastic tissue of human fetal chorionic villus samples (CVSs). After immunoselection of CV cells for SSEA4, FACS analyses showed an increased number of cells positive for OCT-4 and ALP and a small percentage (around 4%) of side population (SP) cells. In the same cell population, RT-PCR indicated the presence of OCT-4, NANOG, and SOX2 transcripts, also typical of stem cells. Depending on the in vitro conditions, a subset of SSEA4+ cells formed colonies resembling hESCs, with limited self renewal ability. At the same time, these cells were able to differentiate in vitro into derivatives of all three germ layers. When inoculated into immunocompromised mice, SSEA4+ cells did not form teratomas but were able to populate depleted hematopoietic tissues. Moreover, after injection into mouse blastocysts, they were incorporated into the inner cell mass and could be traced into several tissues of the adult chimeric mice. Finally, we show that SSEA4+ cells isolated from fetuses affected by Spinal Muscular Atrophy (SMA) can be genetically corrected with high efficiency in culture by Small Fragment Homologous Recombination (SFHR), a gene targeting approach. Taken together, our results indicate that SSEA4+ cells obtained from human CVSs contain a subpopulation of multipotent cells that we propose to name Human Cytotrophoblastic-derived Multipotent Cells (hCTMCs). These cells may be a safe and convenient source of cells for cell-based therapy, as well as an ideal target for in utero fetal gene therapy. PMID- 20025525 TI - Preimplantation development of somatic cell cloned embryos in the common marmoset (Callithrix jacchus). AB - The somatic cell nuclear transfer technique has been applied to various mammals to produce cloned animals; however, a standardized method is not applicable to all species. We aimed here to develop optimum procedures for somatic cell cloning in nonhuman primates, using common marmosets. First, we confirmed that parthenogenetic activation of in vitro matured oocytes was successfully induced by electrical stimulation (three cycles of 150 V/mm, 50 microsec x 2, 20 min intervals), and this condition was applied to the egg activation procedure in the subsequent experiments. Next, nuclear transfer to recipient enucleated oocytes was performed 1 h before, immediately after, or 1 h after egg activation treatment. The highest developmental rate was observed when nuclear transfer was performed 1 h before activation, but none of the cloned embryos developed beyond the eight-cell stage. To investigate the causes of the low developmental potential of cloned embryos, a study was performed to determine whether the presence of metaphase II (MII) chromosome in recipient ooplasm has an effect on developmental potential. As a result, only tetraploid cloned embryos produced by transferring a donor cell into a recipient bearing the MII chromosome developed into blastocysts (66.7%). In contrast, neither parthenogenetic embryos nor cloned embryos (whether diploid or tetraploid) produced using enucleated oocytes developed past the eight-cell stage. These results suggest that MII chromosome, or cytoplasm proximal to the MII chromosome, plays a major role in the development of cloned embryos in common marmosets. PMID- 20025526 TI - Health care-associated Clostridium difficile infection in Canada: patient age and infecting strain type are highly predictive of severe outcome and mortality. AB - BACKGROUND: C. difficile infection (CDI) has become an important and frequent nosocomial infection, often resulting in severe morbidity or death. Severe CDI is more frequently seen among individuals infected with the emerging NAP1/027/BI (NAP1) strain and in the elderly population, but the relative importance of these 2 factors remains unclear. We used a large Canadian database of patients with CDI to explore the interaction between these 2 variables. METHODS: The Canada-wide CDI study, performed in 2005 by the Canadian Nosocomial Infection Surveillance Program (CNISP), was used to analyze the role of infecting strain type and patient age on the severity of CDI. A severe outcome was defined as CDI requiring intensive care unit care, colectomy, or causing death (directly or indirectly) within 30 days after diagnosis. RESULTS: A total of 1008 patients in the CNISP database had both complete clinical data and infecting strain analysis documented. A total of 311 patients (31%) were infected with the NAP1 strain, 83 (28%) were infected with the NAP2/J strain, and the rest were infected with various other types. The proportion of NAP1 infections correlated with the incidence and the severity of CDI when analyzed by province. Thirty-nine (12.5%) of the infections due to the NAP1 strain resulted in a severe outcome, compared with only 41 (5.9%) of infections due to the other types (P < .001). The patient's age was strongly associated with a severe outcome, and patients 60-90 years of age were approximately twice as likely to experience a severe outcome if the infection was due to NAP1, compared with infections due to other types. CONCLUSIONS: Our study confirms the strong age association with infection due to the NAP1 strain and severe CDI. In addition, patients 60-90 years of age infected with NAP1 are approximately twice as likely to die or to experience a severe CDI related outcome, compared with those with non-NAP1 infections. Patients >90 years of age experience high rates of severe CDI, regardless of strain type. PMID- 20025527 TI - Safety and efficacy of intravenous tigecycline in subjects with secondary bacteremia: pooled results from 8 phase III clinical trials. AB - BACKGROUND: Tigecycline is effective in the treatment of complicated skin/skin structure infection (cSSSI), complicated intraabdominal infection (cIAI), and community-acquired bacterial pneumonia (CAP), but its efficacy in subjects with secondary bacteremia is unknown. METHODS: Pooled data from subjects enrolled for treatment of cSSSI, cIAI, or CAP presenting with bacteremia from 7 double-blind and 1 open-label trial of tigecycline compared with vancomycin-aztreonam, imipenem-cilastatin, levofloxacin, vancomycin, or linezolid were analyzed. The primary efficacy end point was the clinical cure rate at the test-of-cure assessment. RESULTS: A total of 170 subjects were identified (91 tigecycline recipients and 79 recipients of the comparator agent). Clinical cure rates were 81.3% and 78.5% for tigecycline and the comparator, respectively (P = .702). Analysis by sex, age, creatinine clearance, infection site, Acute Physiology and Chronic Health Evaluation score, and Fine score demonstrated no significant between-group differences. Clinical cure rates for the most commonly represented pathogens (Staphylococcus aureus, Streptococcus pneumoniae, and gram-negative species) were also not significantly different between treatment groups. No decrease in the rate of cure was found in organisms with increasing tigecycline minimum inhibitory concentrations. Nine subjects treated with tigecycline and 1 subject treated with comparator were found to have persistent bacteremia. No clinically significant differences in safety parameters were identified. CONCLUSIONS: Tigecycline was generally safe and well tolerated in the treatment of secondary bacteremia associated with cSSSI, cIAI, and CAP; cure rates were similar to comparative standard therapies. PMID- 20025528 TI - Reactive arthritis after enteric infections in the United States: the problem of definition. AB - Bacterial enteric infections cause substantial morbidity in the United States both from acute illness and sequelae that follow. Reactive arthritis (ReA) is a poorly defined term that is used to describe a variety of rheumatologic phenomena that may occur after Salmonella, Shigella, Campylobacter, and Yersinia infection, as well as other types of infections (eg, Chlamydia). This review focuses on clinical and epidemiologic investigations of ReA following bacterial enteric infection in the United States. Only 2 population-based studies of ReA following enteric infection have been performed in the United States. ReA following outbreaks of Campylobacter and Yersinia infection has not been studied, and investigations following Shigella and Salmonella outbreaks have focused primarily on the more narrowly defined, but now outdated, concept of "Reiter's syndrome" rather than ReA. Additional epidemiologic studies are needed to determine the burden of illness due to ReA following enteric infection, but a clearer definition of the term is a prerequisite. PMID- 20025529 TI - Double-edged genetic swords and immunity: lesson from CCR5 and beyond. PMID- 20025530 TI - CCR5 deficiency is a risk factor for early clinical manifestations of West Nile virus infection but not for viral transmission. AB - BACKGROUND: West Nile virus (WNV) is a neurotropic flavivirus transmitted to humans by mosquito vectors. Homozygosity for CCR5Delta32, a complete loss-of function mutation in CC chemokine receptor 5 (CCR5), has been previously associated with severe symptomatic WNV infection in patients who present with clinical disease; however, whether it acts at the level of initial infection or in promoting clinical progression is unknown. METHODS: Here, we address this gap in knowledge by comparing CCR5Delta32 distribution among US blood donors identified through a comprehensive blood supply screening program (34,766,863 donations from 2003 through 2008) as either WNV true positive (634 WNV-positive cases) or false positive (422 WNV-negative control participants). All subjects self-reported symptoms occurring during the 2 weeks following blood donation using a standardized questionnaire. RESULTS: No difference was observed in CCR5Delta32 homozygous frequency between the WNV-positive cases and WNV-negative control participants. However, CCR5Delta32 homozygosity was associated in cases but not controls with clinical symptoms consistent with WNV infection (P = .002). CONCLUSIONS: CCR5 deficiency is not a risk factor for WNV infection per se, but it is a risk factor for both early and late clinical manifestations after infection. Thus, CCR5 may function normally to limit disease due to WNV infection in humans. PMID- 20025531 TI - Evaluation of ultraviolet C for disinfection of endocavitary ultrasound transducers persistently contaminated despite probe covers. AB - OBJECTIVE: To determine the rate of bacterial and viral contamination of endocavitary ultrasound probes after endorectal or endovaginal examination with the use of probe covers and to evaluate the antimicrobial efficacy of a disinfection procedure consisting of cleaning with a disinfectant-impregnated towel followed by disinfection with ultraviolet C (UVC) light. METHODS: Endovaginal or endorectal ultrasound examinations were performed for 440 patients in 3 institutions. All probes were covered by a condom or sheath during the examination. For bacterial analysis, 1 swab was applied lengthwise across one half the surface of the probe just after removal of the probe cover. The second swab was similarly applied over the probe immediately after the end of a 2-step process consisting of cleaning with a towel impregnated with a disinfectant spray and a 5-minute UVC disinfection cycle. Swabs were applied onto plates and incubated for 48 hours. The number of colony-forming units was counted, and organisms were identified. A similar protocol was used for viral detection of Epstein-Barr virus, human cytomegalovirus, and human papillomavirus, except that an additional swab was applied along the entire external surface of the probe cover before its removal. Viruses were detected by means of a polymerase chain reaction-based protocol. RESULTS: After removal of probe covers, contamination by pathogenic bacteria was found for 15 (3.4% [95% confidence interval, 2.0%-5.6%]) of 440 probes, and viral genome was detected on 5 (1.5% [95% confidence interval, 0.5%-3.5%]) of 336 probes. After cleaning with a towel impregnated with a disinfectant spray and disinfecting with UVC light, neither bacterial pathogenic flora nor viral genome was recovered from the probe. CONCLUSIONS: Endocavitary ultrasound probes may carry pathogens after removal of covers under routine conditions. A disinfection procedure consisting of cleaning with a disinfectant impregnated towel followed by disinfection with UVC may provide a useful method for disinfecting endocavitary ultrasound probes. PMID- 20025532 TI - Blood culture contamination rates after skin antisepsis with chlorhexidine gluconate versus povidone-iodine in a pediatric emergency department. AB - OBJECTIVE: To determine blood culture contamination rates after skin antisepsis with chlorhexidine, compared with povidone-iodine. DESIGN: Retrospective, quasi experimental study. SETTING: Emergency department of a tertiary care children's hospital. PATIENTS: Children aged 2-36 months with peripheral blood culture results from February 2004 to June 2008. Control patients were children younger than 2 months with peripheral blood culture results. METHODS: Blood culture contamination rates were compared using segmented regression analysis of time series data among 3 patient groups: (1) patients aged 2-36 months during the 26 month preintervention period, in which 10% povidone-iodine was used for skin antisepsis before blood culture; (2) patients aged 2-36 months during the 26 month postintervention period, in which 3% chlorhexidine gluconate was used; and (3) patients younger than 2 months not exposed to the chlorhexidine intervention (ie, the control group). RESULTS: Results from 11,595 eligible blood cultures were reviewed (4,942 from the preintervention group, 4,274 from the postintervention group, and 2,379 from the control group). For children aged 2-36 months, the blood culture contamination rate decreased from 24.81 to 17.19 contaminated cultures per 1,000 cultures (P < .05) after implementation of chlorhexidine. This decrease of 7.62 contaminated cultures per 1,000 cultures (95% confidence interval, -0.781 to -15.16) represented a 30% relative decrease from the preintervention period and was sustained over the entire postintervention period. No change in contamination rate was observed in the control group (P = .337). CONCLUSION: Skin antisepsis with chlorhexidine significantly reduces the blood culture contamination rate among young children, as compared with povidone-iodine. PMID- 20025533 TI - Perichromatin fibrils accumulation in hepatocyte nuclei reveals alterations of pre-mRNA processing during aging. AB - We previously described an unusual accumulation of perichromatin fibrils (PF)-the in situ form of pre-mRNA transcription and early splicing-in hepatocyte nuclei of old rats. Here we have investigated, by immunoelectron microscopy, the nature of such PF, analyzing the presence of transcription, splicing and cleavage factors, polyadenylated RNA, and the incorporation of bromouridine in adult and old rats. Our observations revealed alterations in amount and/or distribution of pre-mRNA transcription, splicing and cleavage factors, as well as of polyadenylated RNA, together with lower bromouridine incorporation in newly transcribed RNA in the hepatocyte nucleoplasm of old rats. Therefore, our data indicate both a decrease in pre-mRNA transcription and a slow down of PF processing and transport during aging. PMID- 20025534 TI - Haplotype-based analysis of MDR1/ABCB1 gene polymorphisms in a Turkish population. AB - The three functional single-nucleotide polymorphisms (SNPs) of the MDR1 gene, C1236T, G2677T/A, and C3435T, exhibit an interpopulation difference. In this study, we analyzed the haplotype frequencies of these three SNPs in 107 unrelated healthy Turkish subjects and compared them with those of other reported populations. We found that C1236T, G2677T/A, and C3435T SNPs are expected to be structured in 10 different haplotypes, with 4 prominent haplotypes T-T-T (33.7%), C-G-C (25.0%), T-G-C (10.9%), and C-T-T (8.7%). There was a statistically significant linkage disequilibrium between all C1236T, G2677T/A, and C3435T SNPs (p < 0.0001); however, our results indicated that only loci 2677 and 3435 show relatively strong linkage disequilibrium (Lewontin's coefficient [D'] = 0.74, Pearson's correlation [r(2)] = 0.47). The haplotype frequency distribution of our study group was found to be significantly different from that in Han Chinese, Uygur Chinese, Kazakh Chinese, Indian, Malay, Japanese, Caucasian, and Ashkenazi Jewish populations (p < 0.0001). The results of this study may contribute to population-specific haplotype data on the MDR1 gene and may serve as a basis for studies on response to P-glycoprotein substrate drugs as well as for future association studies of certain diseases in Turkish population. PMID- 20025535 TI - Characterization of the 5'-flanking region and regulation of transcription of human BAFF-R gene. AB - B-cell activating factor (BAFF) is critical for maintaining the development and homeostasis of B cells. Overexpression of BAFF is associated with autoimmune diseases and malignant B lymphoma. BAFF receptor (BAFF-R) was found to be a specific receptor of BAFF. It not only plays a significant role in splenic B-cell maturation but also works as a major mediator in BAFF-dependent costimulatory response in peripheral B and T cells. Previous studies have demonstrated that BAFF-R is related to several diseases; however, the molecular mechanism of BAFF-R genic transcription has not been clearly defined. The aim of this study was to investigate the transcriptional regulation of the BAFF-R gene. This study was designed to clone and characterize the 5'-regulatory region of the human BAFF-R gene and determine the mechanisms involved in its transcriptional regulation. In addition, the effects of interferon (IFN)-gamma and BAY11-7082 (inhibitor of nuclear factor [NF]-kappaB) on the expression and promoter activity of BAFF-R were examined. The results showed that the sequence between -1420 and +261 could be a core promoter region, and -1562 and -1420 bp harbored a transcriptive silencer. IFN-gamma promoted BAFF-R promoter activity and upregulated BAFF-R mRNA expression. BAY11-7082 (inhibitor of NF-kappaB) exhibited an inhibitory effect on BAFF-R promoter activity and downregulated BAFF-R mRNA expression. Our data provided novel evidence to clarify the mechanism of transcriptional regulation of BAFF-R and illustrated that IFN-gamma and NF-kappaB pathway were involved in regulating BAFF-R expression. Thus some BAFF-R-related diseases might be cured by blocking transcriptional regulation of BAFF-R and reducing its expression. PMID- 20025536 TI - Electrotransfer of cDNA coding for a heterologous prion protein generates autoantibodies against native murine prion protein in wild-type mice. AB - Prion diseases (e.g., Creutzfeldt-Jakob disease in humans) are always fatal neurodegenerative disorders characterized by conversion of the ubiquitous cellular prion protein (PrP(c)) into a pathological conformer. Immunological strategies are considered as promising prophylactic or therapeutic approaches but, unfortunately, vaccination attempts until now have been very disappointing in wild-type animals because of immune tolerance to self PrP(c). Encouraging results have come from recent experiments carried out through genetic immunization (i.e., injection in mice of cDNA coding for murine prion protein [PrP]) or heterologous protein immunization (i.e., injection in mice of PrP from another species), albeit the levels of autoantibodies in wild-type animals remained generally low. Here we investigated whether combining the potential benefits of these two last approaches, namely using genetic immunization with the cDNA coding for a heterologous PrP, could more efficiently break immune tolerance. Wild-type mice were thus vaccinated with cDNA coding for human PrP(c), fused or unfused to a stimulatory T-cell epitope, using or not using electrotransfer of DNA. After three DNA injections, mice receiving electrotransferred DNA developed a strong immune response, oriented toward the humoral Th2 type, characterized not only by high IgG1 and IgG2a antibody titers against the heterologous human PrP(c), but also, as expected, by significant amounts of autoantibodies recognizing the native conformation of murine PrP(c) expressed on cell membranes as revealed by flow cytometry and immunofluorescence. These results hence open the way for investigation of the possible protective effects of anti-PrP(c) autoantibodies in infected mouse models. More generally, our results suggest that this original immunization strategy could be of value for circumventing tolerance to poorly immunogenic proteins. PMID- 20025537 TI - Cytogenetic and antineoplastic effects of modified steroidal alkylators. AB - INTRODUCTION: The aim of this study was to design new potentially antineoplastic agents by combining nitrogen mustard with steroidal skeleton, in an effort to improve specificity and simultaneously to reduce systemic toxicity. The steroidal part is aimed to act as a biological platform enabling the alkylating moiety to approach its site of action by altering its physicochemical properties. MATERIALS AND METHODS: The compounds tested have, as alkylating agents, either p-N,N-bis(2 chloroethyl)aminophenyl-butyrate or p-N,N-bis(2-chloroethyl)aminophenyl-acetate esterified with a modified steroidal nucleus. The four newly synthesized compounds were compared on a molar basis, regarding their ability to induce sister chromatid exchanges and modify proliferation rate indices in cultured human lymphocytes. Life span of BDF1 mice inoculated with L1210 leukemia was also estimated (antileukemic activity). RESULTS: A compound having p-N,N-bis(2 chloroethyl)aminophenyl-acetate as the alkylator and two ketone groups in the steroidal part demonstrated the highest statistically significant enhancement of sister chromatid exchanges and suppression of proliferation rate indices, and also caused significant antineoplastic activity. The other compounds proved less active. CONCLUSION: These results suggest that cytogenetic and antileukemic activity of alkylating steroidal esters depends on the configuration of the whole molecule and the appropriate combination of the alkylator with the steroidal molecule. PMID- 20025538 TI - Induction chemotherapy with weekly paclitaxel administration for anaplastic thyroid carcinoma. AB - BACKGROUND: Anaplastic thyroid carcinoma (ATC) has a very dire prognosis and no effective therapeutic strategies have been established for ATC patients, especially those with stage IVB or IVC. Our objective was to investigate the effectiveness of weekly paclitaxel administered as induction chemotherapy and to establish novel therapeutic strategies for ATC. METHODS: We performed induction chemotherapy by weekly paclitaxel administration for patients with stage IVB (nine patients) and IVC (four patients) disease. Clinical and histological responses were assessed. Overall survival was compared with that of ATC patients with stage IVB (n = 50) and IVC (n = 13) treated without paclitaxel. RESULTS: One patient demonstrated complete response (CR) and two demonstrated partial response (PR) in the stage IVB group (response rate: 33%), and one patient showed PR in stage IVC (response rate: 25%). Curative surgery and adjuvant therapy were performed for four patients with stage IVB, and 32 months after treatment, all the four are alive and free of disease. One CR patient and one stable disease patient were assessed as grade IV (complete remission) and grade III (nearly CR) on histological response, respectively. All four patients with stage IVC died of carcinoma within 8 months. Overall survival of stage IVB patients with induction chemotherapy was better (p = 0.0213) than that without the chemotherapy and also better (p = 0.0467) than those with chemotherapy other than paclitaxel. However, induction chemotherapy did not improve the overall survival of stage IVC patients (p = 0.2002). CONCLUSIONS: Induction chemotherapy by weekly paclitaxel is a promising therapeutic strategy for stage IVB ATC patients. Responders can be expected to achieve long-term survival. We could not get significant difference of overall survival between stage IVC patients with or without weekly induction paclitaxel, although there were some cases that responded well. PMID- 20025539 TI - Diagnosis and treatment of pancreatic metastases of a papillary thyroid carcinoma. AB - BACKGROUND: Apart from regional lymph node metastases, systemic metastases occur sporadically in papillary thyroid carcinomas (PTC). The lung and bones are the most frequent localizations. Additionally known but extremely rare locations are metastases of the skeletal muscles, ovaries, submandibular gland, sphenoidal sinus, brain, adrenals, and, as shown in only two previously published cases to date, the pancreas. SUMMARY: In this article we report about two additional patients with pancreatic metastases from PTC. There is almost no prior experience about therapeutic approaches to this type of metastases. In both patients distant metastases within the pancreas were successfully removed. Postoperative histology confirmed the diagnoses. Supplemental genetic analysis did not demonstrate a BRAF V600E mutation or expression of a RET/PTC1 rearrangement in one case, but revealed a BRAF V600E mutation in the second case. Surgery avoided impending complications maintaining quality of life. One patient had a tumor-specific survival of 42 months. The other patient has occult disease. CONCLUSIONS: Our two patients benefited of a calculated aggressive surgical action. Thus, if low perioperative mortality and morbidity can be warranted, surgical measures are justifiable in selected cases. PMID- 20025540 TI - Clinical and cytological features predictive of malignancy in thyroid follicular neoplasms. AB - BACKGROUND: The preoperative diagnosis of malignancy in nodules suspicious for a follicular neoplasm remains challenging. A number of clinical and cytological parameters have been previously studied; however, none have significantly impacted clinical practice. The aim of this study was to determine predictive characteristics of follicular neoplasms useful for clinical application. METHODS: Four clinical (age, sex, nodule size, solitary nodule) and 17 cytological variables were retrospectively reviewed for 144 patients with a nodule suspicious for follicular neoplasm, diagnosed preoperatively by fine-needle aspiration (FNA), from a single institution over a 2-year period (January 2006 to December 2007). The FNAs were examined by a single, blinded pathologist and compared with final surgical pathology. Significance of clinical and cytological variables was determined by univariate analysis and backward stepwise logistic regression. Odds ratios (ORs) for malignancy, a receiver operating characteristic curve, and predicted probabilities of combined features were determined. RESULTS: There was an 11% incidence of malignancy (16/144). On univariate analysis, nodule size >OR=4.0 cm nears significance (p = 0.054) and 9 of 17 cytological features examined were significantly associated with malignancy. Three variables stay in the final model after performing backward stepwise selection in logistic regression: nodule size (OR = 0.25, p = 0.05), presence of a transgressing vessel (OR = 23, p < 0.0001), and nuclear grooves (OR = 4.3, p = 0.03). The predicted probability of malignancy was 88.4% with the presence of all three variables on preoperative FNA. When the two papillary carcinomas were excluded from the analysis, the presence of nuclear grooves was no longer significant, and anisokaryosis (OR = 12.74, p = 0.005) and presence of nucleolus (OR = 0.11, p = 0.04) were significantly associated with malignancy. Excluding the two papillary thyroid carcinomas, a nodule size >or=4 cm, with a transgressing vessel and anisokaryosis and lacking a nucleolus, has a predicted probability of malignancy of 96.5%. CONCLUSIONS: A combination of larger nodule size, transgressing vessels, and specific nuclear features are predictive of malignancy in patients with follicular neoplasms. These findings enhance our current limited predictive armamentarium and can be used to guide surgical decision making. Further study may result in the inclusion of these variables to the systematic evaluation of follicular neoplasms. PMID- 20025541 TI - Iodide-induced chemokines and genes related to immunological function in cultured human thyroid follicles in the presence of thyrotropin. AB - BACKGROUND: It is well known that iodide exacerbates thyroid function in subclinical hypothyroid patients with autoimmune thyroiditis. To investigate the immunological mechanism of iodine-induced thyroid dysfunction, we studied the effect of iodide in cultured human thyroid follicles, which respond to physiological concentrations of human thyrotropin (TSH) (0.3-10 microU/mL) and maintain the Wolff-Chaikoff effect. MATERIALS AND METHODS: Thyroid follicles obtained from Graves' patients at subtotal thyroidectomy were precultured in medium containing 0.5% fetal calf serum and 10(-8) M iodide for 5 days, and then cultured with the medium containing bovine TSH (30 microU/mL) and low (10(-8)M) or high (10(-5)M) concentrations of iodide. After 3-72 hours of culture, the effect of iodide on thyroid cell mRNA expression was analyzed by microarray and reverse transcriptase-polymerase chain reaction. RESULTS: After 48 hours of culture, iodide nearly doubled the mRNA expression levels of the immunity associated genes (intercellular adhesion molecule-1, transforming growth factor beta 1-induced protein, early growth response gene 1, guanylate-binding protein 1, and annexin A1) and decreased the mRNA expression of sodium-iodide symporter to less than 20%. Further, the mRNA expression levels of chemokines (CCL2, CXCL8, and CXCL14) increased nearly twofold, whereas their receptors did not show any significant response. Real-time polymerase chain reaction analyses confirmed that iodide increased the mRNA expression levels of these genes in a time- and concentration-dependent manner. Immunohistochemical studies revealed that the chemokines were expressed mainly in the thyroid follicular cells in addition to the immune cells. The iodide-induced increase in CCL2 was greater in thyroid follicles obtained from thyroid gland that had been moderately infiltrated with the immunocompetent cells. CONCLUSION: We have demonstrated that iodide stimulates thyroid follicular cells to produce chemokines, particularly CCL2, CXCL8, and CXCL14. These chemokines and intercellular adhesion molecule-1 would attract immunocompetent cells into thyroid gland. These in vitro findings suggest that iodide at high concentrations may induce thyroid dysfunction through not only biochemical but also immunological mechanisms, particularly in patients with autoimmune thyroid disorders. PMID- 20025542 TI - Power in the study of mortality and necrotizing soft tissue infections. PMID- 20025543 TI - Radionuclide imaging of tumor angiogenesis. AB - Angiogenesis is a multistep process regulated by pro- and antiangiogenic factors. In order to grow and metastasize, tumors need a constant supply of oxygen and nutrients. For growth beyond 1-2 mm in size, tumors are dependent on angiogenesis. Inhibition of angiogenesis is a new cancer treatment strategy that is now widely investigated clinically. Researchers have begun to search for objective measures that indicate pharmacologic responses to antiangiogenic drugs. Therefore, there is a great interest in techniques to visualize angiogenesis in growing tumors noninvasively. Several markers have been described that are preferentially expressed on newly formed blood vessels in tumors (alpha(v)beta(3) integrin, vascular endothelial growth factor, and its receptor, prostate-specific membrane antigen) and in the extracellular matrix surrounding newly formed blood vessels (extra domain B of fibronectin, Tenascin-C, matrix metalloproteinases, and Robo-4). Several ligands targeting these markers have been tested as a radiotracer for imaging angiogenesis in tumors. The potential of some of these tracers, such as radiolabeled cyclic RGD peptides and radiolabeled anti-PSMA antibodies, has already been tested in cancer patients, while for markers such as Robo-4, the ligand has not yet been identified. In this review, an overview on the currently used nuclear imaging probes for noninvasive visualization of tumor angiogenesis is given. PMID- 20025544 TI - Glomerular filtration rate after alpha-radioimmunotherapy with 211At-MX35 F(ab')2: a long-term study of renal function in nude mice. AB - Besides bone marrow, the kidneys are often dose-limiting organs in internal radiotherapy. The effects of high-linear energy transfer (LET) radiation on the kidneys after alpha-radioimmunotherapy (alpha-RIT) with the alpha-particle emitter, (211)At, were studied in nude mice by serial measurements of the glomerular filtration rate (GFR). The renal toxicity was evaluated at levels close to the dose limit for the bone marrow and well within the range for therapeutic efficacy on tumors. Astatinated MX35-F(ab')(2) monoclonal antibodies were administered intravenously to nude mice. Both non-tumor-bearing animals and animals bearing subcutaneous xenografts of the human ovarian cancer cell line, OVCAR-3, were used. The animals received approximately 0.4, 0.8, or 1.2 MBq in one, two, or three fractions. The mean absorbed doses to the kidneys ranged from 1.5 to 15 Gy. The renal function was studied by serial GFR measurements, using plasma clearance of (51)Cr-EDTA, up to 67 weeks after the first astatine injection. A dose-dependent effect on GFR was found and at the time interval 8-30 weeks after the first administration of astatine, the absorbed doses causing a 50% decrease in GFR were 16.4 +/- 3.3 and 14.0 +/- 4.1 Gy (mean +/- SEM), tumor- and non-tumor-bearing animals, respectively. The reduction in GFR progressed with time, and at the later time interval, (31-67 weeks) the corresponding absorbed doses were 7.5 +/- 2.4 and 11.3 +/- 2.3 Gy, respectively, suggesting that the effects of radiation on the kidneys were manifested late. Examination of the kidney sections showed histologic changes that were overall subdued. Following alpha-RIT with (211)At-MX35-F(ab')(2) at levels close to the dose limit of severe myelotoxicity, the effects found on renal function were relatively small, with only minor to moderate reductions in GFR. These results suggest that a mean absorbed dose to the kidneys of approximately 10 Gy is acceptable, and that the kidneys would not be the primary dose-limiting organ in systemic alpha-RIT when using (211)At-MX35-F(ab')(2). PMID- 20025545 TI - Lymphocytic toxicity in patients after peptide-receptor radionuclide therapy (PRRT) with 177Lu-DOTATATE and 90Y-DOTATOC. AB - PURPOSE: Peptide-receptor radionuclide therapy (PRRT) with somatostatin analogs is an efficient new tool in patients with neuroendocrine tumors, with low risk of toxicity. Since lymphocytes express somatostatin receptors, the aim of this study was to evaluate lymphocytic toxicity after PRRT. METHODS: From May 2005 to May 2007, 16 patients affected by neuroendocrine tumors received PRRT with (90)Y DOTATOC (9), (177)Lu-DOTATATE (5), or both (2). Absolute count, percentage of leukocytes and lymphocytes, and lymphoid subsets (B, T, and NK) were tested at baseline and until 90 days after treatment. RESULTS: A significant lymphoid toxicity (G2-3), mainly affecting B-cells, was observed. It was particularly evident after (90)Y-DOTATOC. Toxicity resulted in being transient and resolved completely at the end of the follow-up (90 days). CONCLUSION: Lymphocyte toxicity in PRRT is mainly due to the selective targeting on B-cells. The relative sparing of T-lymphocytes could explain the absence of clinical side-effects in these patients, such as increased risk of infections. These findings open interesting perspectives in the treatment of B-cell lymphoproliferative disorders. PMID- 20025546 TI - Signaling defects and functional impairment in T-cells from cervical cancer patients. AB - The ability of T-lymphocytes to recognize antigens and transduce signals to the nucleus successfully is a key component in the initiation and maintenance of an immune response. The present study addressed the expression status of the signal transducing proteins in relation to the immune impairment in cervical cancer patients. Immune response was measured by evaluating lymphocyte subpopulations CD3(+), CD4(+), and CD8(+), using flowcytometry, natural killer cell activity, using the single-cell cytotoxicity assay, lymphocyte function, using mitogenic response to PHA and T-cell activation following anti-CD3 stimulation, and production of IL-2. Expression of the T-cell signal transduction proteins, TCR zeta, CD3-epsilon, zap-70, p(56)lck, PKC, NFkappabeta p50, Rel-A, Rel-B, and c rel, was evaluated by using Western blot assay. A generalized depression of the immune response with respect to the different parameters evaluated was observed. Exogenous interleukin-2 (IL-2) could increase the response in all the controls and in 30% of the patients to different degrees varying from 10% to 90%. Low levels of the signaling molecules (TCR-zeta, CD3-epsilon, zap-70, p(56)lck, and PKC) and impairment in the transduction of NFkappabeta components (p50, Rel-A, Rel-B, and c-rel) to the nuclei were observed in these lymphocytes. Decreased CD4(+)/CD8(+) ratio with an increase in suppressor cells, reduced lymphocyte proliferation, and production of IL-2 suggest a defective immune regulation in cervical cancer. Impairment in the translocation of NFkappabeta p50, Rel-A, and Rel-B to the nucleus and the reduced levels of signal-transducing proteins might be responsible for the decreased production of IL-2 and immune impairment in cervical cancer patients. PMID- 20025547 TI - Resveratrol protects against Cisplatin-induced cardiotoxicity by alleviating oxidative damage. AB - The clinical use of cisplatin, a potent antineoplastic agent, is limited by its severe adverse effects. The present study was designed to evaluate the effects of resveratrol on cisplatin-induced cardiac injury. Resveratrol is a potent free radical scavenger. In the present study, we tested whether resveratrol would prevent cisplatin-induced cardiotoxicity in rats. Plasma-enzyme activities and histologic myocardial changes were examined. The anticancer role of resveratrol and/or cisplatin were measured by MTT. Our data showed that cisplatin led to cardiac-function deterioration, myocardial injury, increased lactate dehydrogenase, creatine kinase, malondialdehyde activities, and decreased activities of superoxide dismutase, glutathione, glutathione peroxidase, and catalase. Treatment with resveratrol effectively hindered the adverse effects of cisplatin in a dose-dependent manner, such as myocardial injury and impaired heart function. An in vitro cytotoxic study showed that resveratrol could increase the antineoplastic activity of cisplatin to A549 adenocarcinoma cells. All the above lines of evidence suggest that resveratrol protects cardiomyocytes from cisplatin-induced cardiotoxicity via the suppression of oxidative stress. PMID- 20025548 TI - Anti-inflammatory effects of tanshinone IIA on radiation-induced microglia BV-2 cells inflammatory response. AB - AIM: The aim of this study was to explore the inhibitory effects of Tanshinone II(A) on the production of proinflammation cytokines in radiation-stimulated microglia. METHODS: Microglia cells were treated with 2, 4, 8, 16, and 32 Gy of irradiation or sham-irradiated in the presence or absence of 1.0 microg/mL of Tanshinone II(A). The effects of Tanshinone II(A) on radiation-induced proinflammatory cytokines were evaluated by real-time polymerase chain reaction; the expression level of nuclear factor (NF-kappabeta) p65 in cytoplasm and nucleus was measured by Western blot. Immunofluorescence staining and confocal microscopy analysis were applied to detect the expression of gamma-H2AX and p65 postirradiation. RESULTS: Radiation-induced release of proinflammatory cytokines in BV-2 cells was detectable after irradiation. Tanshinone II(A) decreased the radiation-induced release of proinflammatory cytokines. Further, Western blotting showed that Tanshinone II(A) could attenuate the nuclear translocation of (NF kappabeta) p65 submit postirradiation. Immunofluorescence staining showed gamma H2AX foci formation with p65 translocation into the nucleus postirradiation. CONCLUSIONS: Our data indicated that Tanshinone II(A) exerts anti-inflammatory properties by suppressing the transcription of proinflammatory cytokine genes that might be associated with the NF-kappabeta signaling pathway. It is postulated that irradiation causes immediate cellular reaction, and that double strand breaks trigger the molecular response that leads to NF-kappabeta pathway activation. PMID- 20025549 TI - HDAC inhibitor, valproic acid, induces p53-dependent radiosensitization of colon cancer cells. AB - Agents that inhibit histone deacetylases (HDAC inhibitors) have been shown to enhance radiation response. The aim of this study was to evaluate the effects of low, minimally cytotoxic concentrations of the HDAC inhibitor, valproic acid (VPA), on radiation response of colorectal cancer cells. Cell lines LS174T and an isogenic pair of HCT116, which differed only for the presence of wild-type p53, were exposed to ionizing radiation (IR) alone, VPA alone, or the combination. Clonogenic survival, gamma-H2AX induction, apoptosis, changes in mitochondrial membrane potential, and mitochondrial levels of p53 and Bcl-2 family proteins were assessed. In vivo studies monitored tumor growth suppression after therapy in mice bearing HCT116/p53(+/+) and HCT116/p53(-/-) tumor xenografts. VPA led to radiosensitization, which was dependent on p53 status. A decrease in clonogenic survival, an increase in apoptosis, and an increase in levels of gamma-H2AX were observed after VPA+IR, compared to IR alone, in wild-type p53 cells (LS174T and HCT116/p53(+/+)), as opposed to p53 null cells (HCT116/p53(-/-)). Exposure to VPA resulted in enhancement of IR-induced mitochondrial localizations of Bax and Bcl xL, mitochondrial membrane potential, and cytochrome c release only in wild-type p53 cell lines. VPA also enhanced tumor growth suppression after IR only in wild type p53 xenografts. These data suggest that VPA may have an important role in enhancing radiotherapy response in colorectal cancer, particularly in tumors with the wild-type p53 genotype. PMID- 20025550 TI - Dosimetric study of Cs-131, I-125, and Pd-103 seeds for permanent prostate brachytherapy. AB - As a well-established single-modality approach for early-stage prostate cancer, transperineal interstitial permanent prostate brachytherapy (TIPPB) has gained increasing popularity due to its favorable clinical results. Currently, three isotopes, namely Cs-131, I-125, and Pd-103, are commercially available for TIPPB. This is the first study to systematically explore the dosimetric difference of these three isotopes for TIPPB. In total, 25 patients with T1-T2c prostate cancer previously implanted with I-125 seeds were randomly selected and replanned with Cs-131, I-125, and Pd-103 seeds to the prescription doses of 115, 145, and 125 Gy, respectively. The planning goals attempted were prostate V(p)100 approximately 95%, D(p)90 >or= 100%, and prostatic urethra D(u)10 7.20 (controls) after birth. METHODS: Spectral information about fetal beat-to-beat heart rate, calculated from direct fetal electrocardiogram registrations, was obtained by using a short-time Fourier transform. MAIN OUTCOME MEASURES: Absolute power and normalised power in the low frequency and high-frequency bands. RESULTS: No differences were found between fetuses with and without acidaemia in absolute low or high frequency power (P = 0.2 and P = 0.3, respectively). During the last 30 minutes of labour, acidaemic fetuses had significantly increased normalised low-frequency power (P = 0.01) and decreased normalised high-frequency power (P = 0.03) compared with non-acidaemic fetuses. These differences were not observed from 3 to 2 hours before birth (P = 0.7 and P = 0.9, respectively). CONCLUSION: The autonomic nervous system of human fetuses at term responds adequately to severe stress during labour. Normalised low and high frequency power of heart-rate variability might be able to discriminate between normal and abnormal fetal condition. PMID- 20025621 TI - Are language and social communication intact in children with congenital visual impairment at school age? AB - BACKGROUND: Development of children with congenital visual impairment (VI) has been associated with vulnerable socio-communicative outcomes often bearing striking similarities to those of sighted children with autism.(1) To date, very little is known about language and social communication in children with VI of normal intelligence. METHODS: We examined the presentation of language and social communication of 15 children with VI and normal-range verbal intelligence, age 6 12 years, using a standardised language assessment and parental reports of everyday social and communicative behaviours. Their profiles were compared to those of typically developing sighted children of similar age and verbal ability. RESULTS: Compared to their sighted peers, and relative to their own good and potentially superior structural language skills, children with VI showed significantly poorer use of language for social purposes. Pragmatic language weaknesses were a part of a broader socio-communicative profile of difficulties, present in a substantial proportion of these children and consistent with the pattern found in sighted children with autism. CONCLUSIONS: There are ongoing socio-communicative and pragmatic language difficulties in children with congenital VI at school age, despite their good intellectual abilities and advanced linguistic skills. Further research is required to unpack the underlying causes and factors maintaining this vulnerability in such children. PMID- 20025620 TI - Incentive-elicited mesolimbic activation and externalizing symptomatology in adolescents. AB - BACKGROUND: Opponent-process theories of externalizing disorders (ExD) attribute them to some combination of overactive reward processing systems and/or underactive behavior inhibition systems. Reward processing has been indexed by recruitment of incentive-motivational neurocircuitry of the ventral striatum (VS), including nucleus accumbens (NAcc). METHODS: We used functional magnetic resonance imaging (fMRI) with an incentive task to determine whether externalizing symptomatology in adolescence is correlated with an enhanced VS recruitment by cues for rewards, or by deliveries of rewards. Twelve community recruited adolescents with externalizing disorders (AED) and 12 age/gender matched controls responded to targets to win or avoid losing $0, $0.20, $1, $5, or an unknown amount (ranging from $0.20 to $5). RESULTS: Cues to respond for rewards activated the NAcc (relative to cues for no incentive), in both subject groups similarly, with greatest NAcc recruitment by cues for the largest reward. Loss-anticipatory NAcc signal increase was detected in a volume-of-interest analysis - but this increase occurred only in trials when subjects hit the target. Relative to controls, AED showed significantly elevated NAcc activation by a linear contrast between reward notification versus notification of failure to win reward. In a post hoc reanalysis, VS and pregenual anterior cingulate activation by the reward versus non-reward outcome contrast also directly correlated with Child Behavior Checklist (CBCL) Externalizing total scores (across all subjects) in lieu of a binary diagnosis. Finally, both groups showed right insula activation by loss notifications (contrasted with avoided losses). CONCLUSIONS: Externalizing behavior, whether assessed dimensionally with a questionnaire, or in the form of a diagnostic categorization, is associated with an exaggerated limbic response to outcomes of reward-directed behavior. This could be a neurobiological signature of the behavioral sensitivity to laboratory reward delivery that is characteristic of children with externalizing symptomatology. Of interest is future research on incentive-motivational processing in more severe, clinically referred AED. PMID- 20025622 TI - Neural correlates of verbal and nonverbal semantic integration in children with autism spectrum disorders. AB - BACKGROUND: Autism is a pervasive developmental disorder characterized by deficits in social-emotional, social-communicative, and language skills. Behavioral and neuroimaging studies have found that children with autism spectrum disorders (ASD) evidence abnormalities in semantic processing, with particular difficulties in verbal comprehension. However, it is not known whether these semantic deficits are confined to the verbal domain or represent a more general problem with semantic processing. The focus of the current study was to investigate verbal and meaningful nonverbal semantic processing in high functioning children with autism (mean age = 5.8 years) using event-related potentials (ERPs). METHOD: ERPs were recorded while children attended to semantically matching and mismatching picture-word and picture-environmental sound pairs. RESULTS: ERPs of typically developing children exhibited evidence of semantic incongruency detection in both the word and environmental sound conditions, as indexed by elicitation of an N400 effect. In contrast, children with ASD showed an N400 effect in the environmental sound condition but not in the word condition. CONCLUSIONS: These results provide evidence for a deficiency in the automatic activation of semantic representations in children with ASD, and suggest that this deficit is somewhat more selective to, or more severe in, the verbal than the nonverbal domain. PMID- 20025625 TI - Choosing the appropriate catheter to cannulate anomalous right coronary arteries. PMID- 20025627 TI - Nociceptin/orphanin FQ peptide in hypothalamic neurones associated with the control of feeding behaviour. AB - Nociceptin/orphanin FQ (N/OFQ), an endogenous peptide agonist of the opioid N/OFQ receptor, has been implicated in the regulation of energy balance. In the present study, we have used immunohistochemistry to investigate the cellular localisation and colocalisation of N/OFQ-immunoreactive cell bodies in hypothalamic regions containing neurones producing orexigenic or anorexigenic transmitters. In colchicine-treated rats, N/OFQ immunoreactivity was demonstrated in many cell bodies of the arcuate nucleus (Arc), paraventricular nucleus (PVN) and lateral hypothalamic area (LHA). Double-labelling revealed that N/OFQ was present in some neurones located in the ventrolateral part of the Arc producing pro opiomelanocortin, as shown by the presence of the anorexigenic peptides alpha melanocyte-stimulating hormone (alpha-MSH) and cocaine- and amphetamine-regulated transcript and, occasionally, in single neurones of the ventrolateral Arc producing orexigenic agouti-related peptide, but not neuropeptide Y. N/OFQ immunoreactivity was also demonstrated in a few tyrosine hydroxylase- or dynorphin (DYN)-containing neurones in the dorsomedial part of the Arc. In the parvocellular PVN, N/OFQ was demonstrated in some thyrotrophin-releasing hormone- or DYN-, but not corticotrophin-releasing hormone-containing neurones. Most N/OFQ immunoreactive neurones in the LHA contained orexin- and DYN, but not melanin concentrating hormone. The results obtained, demonstrating the presence of N/OFQ in some alpha-MSH- and in many orexin-containing neurones, suggest a functional relationship between these neuropeptides and N/OFQ in the control of feeding behaviour and body weight. PMID- 20025628 TI - Oestrogen receptor beta is involved in the actions of oestrogens in the brain for affective behaviour, but not trophic effects in peripheral tissues. AB - The steroid, 17beta-oestradiol (E(2)) has pervasive psychological and physical effects throughout the lifespan. The question arises as to whether there are divergent oestrogen receptor (ER)-mediated mechanisms for these effects in the central nervous system (CNS) and periphery. This review focuses on results of studies using a whole animal model (i.e. female rats and mice) to investigate the relative effects and mechanisms of oestrogens in the CNS and the periphery. By using this approach, it has been possible to differentiate the enhancing effects of E(2) on behavioural processes mediated by the hippocampus, such as affective behaviour, and the trophic effects that increase tumourigenesis and uterine growth. Studies using pharmacological manipulations and knockout mice suggest that a likely mechanism underlying the beneficial effects of E(2) for hippocampal function (but not proliferative effects in the body) involves actions at ERbeta, changes in cell cycle/division (e.g. cyclin D1) and/or histone modifications. Thus, it may be possible to differentiate the beneficial effects of oestrogens through ERbeta, particularly in the CNS, from the negative proliferative effects on peripheral, E(2)-sensitive tissues. PMID- 20025629 TI - Effect of urotensin II on PC12 rat pheochromocytoma cells. AB - Urotensin II (U-II), initially identified as a cyclic peptide from fish urophysis, acts both as a strong vasoconstrictor and vasodilator in the vasculature via its receptor, G-protein coupled receptor 14. In addition, U-II and its receptor are co-expressed in the adrenal medulla, as well as in human pheochromocytomas, suggesting that this peptide may have some function in chromaffin cells. However, the precise role of U-II in these cells is unknown. In the present study, we initially demonstrate that U-II and its receptors mRNA are co-expressed in the rat pheochromocytoma cell line PC12. Moreover, U-II has not effect on tyrosine hydroxylase (TH), the rate-limiting enzyme involved in the biosynthesis of catecholamine, in terms of enzyme activity or at the mRNA level. However, U-II does induce an increase in the phosphorylation of TH specifically at Ser31 without affecting phosphorylation at the two other sites (Ser19 and Ser40). U-II also markedly activates extracellular signal-regulated kinases (ERKs) and p38, but not Jun N-terminal kinase. Blockade of the epidermal growth factor (EGF) receptor by AG1478 significantly reduces activation of ERK, suggesting that EGF receptor transactivation could act upstream of the ERK pathway in PC12 cells. Furthermore, U-II significantly increases dopamine secretion from PC12 cells. Finally, we show that U-II induced significant DNA synthesis in a ERKs and P38 mitogen-activated protein kinase-dependent manner. The results obtained indicate that U-II may exert its effects as a neuromodulator in chromaffin cells. PMID- 20025630 TI - Developmental iodine deficiency and hypothyroidism impair neural development, up regulate caveolin-1 and down-regulate synaptophysin in rat hippocampus. AB - Developmental iodine deficiency leads to inadequate thyroid hormone, which damages the hippocampus. In the present study, we implicate hippocampal caveolin 1 and synaptophysin in developmental iodine deficiency and hypothyroidism. Two developmental rat models were established: pregnant rats were administered either an iodine-deficient diet or propylthiouracil (PTU)-adulterated (5 p.p.m. or 15 p.p.m.) drinking water from gestational day 6 until postnatal day (PN) 28. Nissl staining and the levels of caveolin-1 and synaptophysin in several hippocampal subregions were assessed on PN14, PN21, PN28 and PN42. The results obtained show that surviving cells in the iodine-deficient and PTU-treated rats were lower than in controls. Up-regulation of caveolin-1 and down-regulation of synaptophysin were observed in the iodine-deficient and PTU-treated rats. Our findings implicate decreases in the number of surviving cells and alterations in the levels of caveolin-1 and synaptophysin in the impairments in neural development induced by developmental iodine deficiency and hypothyroidism. PMID- 20025631 TI - Glucocorticoid suppression of intraovarian levels of prostaglandins and plasminogen activator activity at ovulation in the rat ovary. AB - AIM: Ovulation is a local physiological inflammatory process with active participation of inflammatory mediators and immune cells. To prevent extensive inflammatory injury to the follicle at ovulation there is also a local anti inflammatory system at ovulation, converting the inactive glucocorticoid cortisone to the more potent cortisol. The aim of this study was to examine the effects of the potent glucocorticoid analogue, dexamethasone (DEX), on ovulation rate and the ovarian production of the ovulatory mediators prostaglandins (PG) and plasminogen activators (PA). METHODS: DEX (0.3, 3, or 100 microM) was administered to an in vitro rat ovarian perfusion system prior to the addition of an ovulation-inducing dose of luteinizing hormone (LH) and 3-isobutyl-1 methylxanthine (IBMX). Control ovaries were perfused only with LH + IBMX. Each perfusion experiment extended over 20 h with ovulation occurring in vitro around 12-15 h after hormonal stimulation. In a second set of perfusion experiments, extending over 10 h, the tissue levels of PG and PA activity in the ovary were evaluated at a time 2-5 h before anticipated ovulation. RESULTS: The median numbers of ovulated oocytes in the groups with DEX of 0.3, 3, and 100 microM were 17.0, 8.5 and 11.0 per treated ovary, respectively. These numbers were not different from those of LH + IBMX-controls (12.5). DEX (100 microM) suppressed tissue levels of PGE(2) and PA activity and decreased (DEX 3 microM, 100 microM) estradiol levels in the perfusion media. CONCLUSION: These results indicate that certain degrees of suppression of PG, PA activity, and estradiol are not sufficient to modulate ovulation rate and/or that glucocorticoids may positively modulate other mediator pathways that exert inhibitory influence on ovulation. PMID- 20025632 TI - Transcervical hysteroscopic sterilization using cyanoacrylate: a long-term experimental study on sheep. AB - AIM: We investigated the transcervical hysteroscopy application of n-butyl-2 cyanoacrylate in the uterine tube lumen of a non-rodent animals (sheep) with fallopian tube dimensions similar those in humans. METHODS: Standard hysteroscopic procedures were performed on female Texel sheep (n = 26). The right and left ostia were identified. For each ewe, a urethral catheter (5Fr) was used for the delivery of 0.5 mL of saline or an equal volume of n-butyl-2 cyanoacrylate into the uterine tube. Following the procedure, ewes were housed with males of proven fertility for 90 days (equivalent to 5.5 estrous cycles). Postmortem (dye and burst pressure) and in vivo (hysterosalpingogram) testing for tube patency were both performed 90 days and 180 days following the procedure. RESULTS: All animals receiving the saline treatment became pregnant. Gross inspection of uterine tubes following n-butyl-2-cyanoacrylate treatment revealed no visceral adhesions or fibrosis. However, postmortem testing revealed total obstruction within the fallopian tubes. This was confirmed by hysterosalpingogram, in that iodine contrast did not escape into the abdominal cavity. CONCLUSION: The cyanoacrylate promoted a reliable fallopian tube obstruction without fibrosis in an animal model exhibiting a similar tube diameter to that found in women. The technique can be evaluated for efficacy in vivo using hysterosalpingography. PMID- 20025633 TI - Long-term use of dienogest for the treatment of endometriosis. AB - AIM: To investigate the safety and efficacy of 52 weeks of dienogest treatment in patients with endometriosis. METHODS: One hundred and thirty-five patients with endometriosis received 2 mg of dienogest orally each day for 52 weeks. Adverse drug reactions and bone density were evaluated. Global improvement was assessed based on the changes in severity categories of five subjective symptoms during non-menstruation (lower abdominal pain, lumbago, dyschezia, dyspareunia, and pain on vaginal examination) and two objective findings (induration involving the pouch of Douglas and limited uterine mobility). RESULTS: The most common adverse drug reactions included metrorrhagia (71.9%), headaches (18.5%), and constipation (10.4%). No clinically significant changes were noted in the incidence or severity of reactions associated with the course of the treatment period (52 weeks). Changes from the baseline bone mineral density of the lumbar spine measured by dual-energy X-ray absorptiometry were -1.6 +/- 2.4% and -1.7 +/- 2.2% (mean +/- standard deviation) at 24 and 52 weeks, respectively, which were statistically significant decreases; however, there was no cumulative decrease. The proportions of patients assessed as marked or moderate improvement in terms of global improvement were 72.5% (95/131 cases) at 24 weeks and 90.6% (106/117 cases) at 52 weeks. CONCLUSION: The long-term effect of dienogest on bone mineral density was slight, whereas the efficacy increased cumulatively. PMID- 20025634 TI - Tension-free vaginal tape-obturator procedure for treatment of severe urodynamic stress incontinence: subjective and objective outcomes during 2 years of follow up. AB - AIM: To report our experience of the tension-free vaginal tape-obturator (TVT-O) procedure for the treatment of severe female urodymanic stress incontinence (USI) during 2 years of follow-up. METHODS: Between January 2005 and June 2007, 67 women with severe USI who underwent the TVT-O procedure were enrolled in the study. Pelvic examination, pad test, urodynamic study, and quality of life (QoL) assessment were performed pre- and post-operatively. Regular follow-up was arranged for analysis. RESULTS: The median duration of follow-up was 24 months. The objective cure rate was 76.2%. The postoperative pad test revealed significant reduction of leakage. The subjective cure rate was 83.5%, and improvement occurred in 14.9%. The postoperative QoL showed significant improvement. One (1.5%) extrusion of tape occurred and three cases of de novo detrusor overactivity were detected. Three patients (4.4%) had immediate postoperative urine retention. No major complications occurred. CONCLUSION: Our results show that the TVT-O procedure is an effective and safe surgical procedure for severe female USI with satisfactory outcomes, significant improvement in quality of life, and few complications during 2 years of follow-up. PMID- 20025635 TI - Soy extract phytoestrogens with high dose of isoflavones for menopausal symptoms. AB - AIM: The aim of the present study was to assess the efficacy and safety of a standardized compound based on an extract of soy phytoestrogens, with high doses of isoflavones in the management of menopausal hot flushes. METHODS: A total of 180 women aged 40-65 years with a minimum of five moderate-to-severe hot flushes in the last 7 days at baseline and absence of menstruation for at least 6 months participated in a 12-week prospective, randomized, double-blind, placebo controlled multicenter trial. After a 2-week run-in period, women received one tablet a day of 80 mg isoflavones (corresponding to 60 mg of genistein) or a matching placebo. RESULTS: The mean daily number of moderate-to-severe hot flushes decreased in both study groups, but the reduction was greater in the isoflavones arm at 6 (36.2%) and 12 weeks (41.2%) than in the placebo arm (24.0% at 6 weeks, 29.3% at 12 weeks), with a difference of 1.1 (95% CI [-2.0 to -0.06]) (P = 0.038) at 6 weeks and 1.1 (95% CI [-2.05 to -0.15]) (P = 0.023) at 12 weeks. Similar findings were obtained for hot flushes of any intensity. The Kupperman index decreased in both study groups. Relief of hot flushes was greater when time to menopause was >or=12 months and in cases of BMI >or=27 kg/m(2). CONCLUSION: In daily practice conditions, high doses of isoflavones, particularly genistein, can be used for the management of hot flushes in postmenopausal women not treated with hormone replacement therapy due to their superior efficacy to placebo and very good safety profile. PMID- 20025636 TI - Efficacy of octyl methoxycinnamate in preventing postoperative peritoneal adhesions: an experimental model. AB - AIM: To evaluate the effect of octyl methoxycinnamate on preventing postoperative peritoneal adhesions (PPA). METHODS: Forty outbred Wistar albino female rats (mean weight, 230 +/- 25 g; mean age, 6 months) were divided into four equal groups. In Group 1, 0.1 mL octyl methoxycinnamate was injected into the peritoneal cavity. In Group 2, only an adhesion model was generated. In Group 3, an adhesion model was generated, and then this area was covered with 0.1 mL octyl methoxycinnamate. In Group 4, an adhesion model was generated after the area was covered with 0.1 mL octyl methoxycinnamate. All rats were sacrificed on postoperative day 10, and adhesions were scored both macroscopically and microscopically. RESULTS: The macroscopic adhesion score of Group 2 differed significantly from the scores in Groups 3 and 4 (P < 0.05), but there was no difference between Groups 3 and 4 (P > 0.05). Microscopic evaluation showed that the mean histopathological fibrosis score of Group 3 differed significantly from the score in Group 4 (P = 0.029). CONCLUSION: Octyl methoxycinnamate covering peritoneal surfaces effectively decreases adhesion formation, but the effect is more pronounced if octyl methoxycinnamate is applied before trauma induction. These results indicate that covering of peritoneal surfaces with viscous liquids, such as octyl methoxycinnamate, which have no toxic effects on vital tissues and especially on peritoneal mesothelial cells, prior to peritoneal trauma (e.g. laparotomy) may decrease PPA formation by preventing or decreasing the trauma. PMID- 20025637 TI - Cerebral venous sinus thrombosis in early pregnancy: an unusual presentation of primary antiphospholipid syndrome. AB - Cerebral venous sinus thrombosis, an unusual presentation of antiphospholipid syndrome, is a rare condition in pregnancy, particularly in the first trimester. The authors of this case report present the case of a 20-year-old woman with sudden onset of headaches and hemiparesis in the first trimester of pregnancy. She underwent a computed tomography scan and magnetic resonance imaging. The image findings and the autoimmune serum test were consistent with a cerebral venous sinus thrombosis complicated by primary antiphospholipid syndrome. The patient's hemiparesis improved with subcutaneous low molecular weight heparin and oral aspirin. In the third trimester of pregnancy, she developed seizures that were controlled by antiepileptic drugs. She delivered a healthy baby at 37 weeks' gestation without immediate or late postpartum complications. PMID- 20025638 TI - Prenatal spontaneous disruption of the dividing membrane in monochorionic diamniotic twins detected at the time of fetoscopic laser photocoagulation. AB - Spontaneous antepartum rupture of the dividing membrane occurring in monochorionic diamniotic twins (MD twin) is an extremely rare complication and difficult to diagnose prenatally. We present a case of pseudo-monoamniotic twins derived from an MD twin gestation, which was suspected by ultrasound and was confirmed by antepartum fetoscopy. A 28-year-old woman, gravida 1, para 1 at 24 weeks of gestation was referred because of suspected polyhydroamnios in an MD twin. Ultrasound suggested twin-twin transfusion syndrome stage III, spontaneous rupture of the dividing membranes and cord entanglement. Fetoscopic laser photocoagulation (FLP) was performed using the Nd:YAG laser on 12 placental vascular connections. Fetoscopy revealed the spontaneous rupture of the dividing membrane and cord entanglement. The remainder of the pregnancy was managed as a monoamniotic twin gestation. Elective cesarean section was performed at 32 weeks of gestation following antenatal steroids and concordantly grown healthy male infants were delivered. PMID- 20025639 TI - Malignant peripheral nerve sheath tumor of the uterine cervix expressing both S 100 protein and HMB-45. AB - A 50-year-old woman presented with a large cervical polypoid mass. Grossly, the mass occupied a substantial proportion of the cervical canal, measuring 6 cm. Histologically, the mass showed a spindle cell malignancy arranged in large fascicles that penetrated deeply into the fibromuscular wall of the cervix. The spindle cells were immunoreactive for both S-100 protein and HMB-45 antigen, but were negative for Melan-A. Electron microscopy showed that cytoplasmic processes of the spindle to oval tumor cells contained microtubules and were lined by basal lamina and abundant intercellular collagen spacing with no melanosomes in any stage. As far as we are aware, this is the ninth reported case of cervical malignant peripheral nerve sheath tumor (MPNST), and the second reported case of MPNST expressing HMB-45 antigen. PMID- 20025640 TI - Malaria: therapeutic implications of melatonin. AB - Malaria, which infects more than 300 million people annually, is a serious disease. Epidemiological surveys indicate that of those who are affected, malaria will claim the lives of more than one million individuals, mostly children. There is evidence that the synchronous maturation of Plasmodium falciparum, the parasite that causes a severe form of malaria in humans and Plasmodium chabaudi, responsible for rodent malaria, could be linked to circadian changes in melatonin concentration. In vitro melatonin stimulates the growth and development of P. falciparum through the activation of specific melatonin receptors coupled to phospholipase-C activation and the concomitant increase of intracellular Ca2+. The Ca2+ signaling pathway is important to stimulate parasite transition from the trophozoite to the schizont stage, the final stage of intraerythrocytic cycle, thus promoting the rise of parasitemia. Either pinealectomy or the administration of the melatonin receptor blocking agent luzindole desynchronizes the parasitic cell cycle. Therefore, the use of melatonin antagonists could be a novel therapeutic approach for controlling the disease. On the other hand, the complexity of melatonin's action in malaria is underscored by the demonstration that treatment with high doses of melatonin is actually beneficial for inhibiting apoptosis and liver damage resulting from the oxidative stress in malaria. The possibility that the coordinated administration of melatonin antagonists (to impair the melatonin signal that synchronizes P. falciparum) and of melatonin in doses high enough to decrease oxidative damage could be a novel approach in malaria treatment is discussed. PMID- 20025642 TI - Melatonin improves sleep and reduces nitrite in the exhaled breath condensate in cystic fibrosis--a randomized, double-blind placebo-controlled study. AB - Cystic fibrosis (CF) is a chronic progressive disorder characterized by repeated episodes of respiratory infection. Impaired sleep is common in CF leading to reduced quality of life. Melatonin, a secretory product of the pineal gland, has an important function in the synchronization of circadian rhythms, including the sleep-wake cycle, and has been shown to possess significant anti-oxidant properties. To evaluate the effects of exogenous melatonin on sleep and inflammation and oxidative stress markers in CF, a randomized double-blind, placebo-controlled study initially involving 20 patients with CF was conducted. One individual failed to conclude the study. All subjects were clinically stable when studied and without recent infectious exacerbation or hospitalization in the last 30 days. Groups were randomized for placebo (n = 10; mean age 12.1 +/- 6.0) or 3 mg melatonin (n = 9; mean age 16.6 +/- 8.26) for 21 days. Actigraphy was performed for 6 days before the start of medication and in the third week (days 14-20) of treatment. Isoprostane and nitrite levels were determined in exhaled breath condensate (EBC) at baseline (day 0) and after treatment (day 21). Melatonin improved sleep efficiency (P = 0.01) and tended to improve sleep latency (P = 0.08). Melatonin reduced EBC nitrite (P = 0.01) but not isoprostane. In summary, melatonin administration reduces nitrite levels in EBC and improves sleep measures in clinically stable CF patients. The failure of melatonin to reduce isoprostane levels may have been a result of the low dose of melatonin used as a treatment. PMID- 20025641 TI - Sirtuins, melatonin and circadian rhythms: building a bridge between aging and cancer. AB - Histone deacetylases (HDAC) have been under intense scientific investigation for a number of years. However, only recently the unique class III HDAC, sirtuins, have gained increasing investigational momentum. Originally linked to longevity in yeast, sirtuins and more specifically, SIRT1 have been implicated in numerous biological processes having both protective and/or detrimental effects. SIRT1 appears to play a critical role in the process of carcinogenesis, especially in age-related neoplasms. Similarly, alterations in circadian rhythms as well as production of the pineal hormone melatonin have been linked to aging and cancer risk. Melatonin has been found act as a differentiating agent in some cancer cells and to lower their invasive and metastatic status. In addition, melatonin synthesis and release occurs in a circadian rhythm fashion and it has been linked to the core circadian machinery genes (Clock, Bmal1, Periods, and Cryptochromes). Melatonin has also been associated with chronotherapy, the timely administration of chemotherapy agents to optimize trends in biological cycles. Interestingly, a recent set of studies have linked SIRT1 to the circadian rhythm machinery through direct deacetylation activity as well as through the nicotinamide adenine dinucleotide (NAD(+)) salvage pathway. In this review, we provide evidence for a possible connection between sirtuins, melatonin, and the circadian rhythm circuitry and their implications in aging, chronomodulation, and cancer. PMID- 20025643 TI - Synergistic antitumor effect of melatonin with several chemotherapeutic drugs on human Ewing sarcoma cancer cells: potentiation of the extrinsic apoptotic pathway. AB - Ewing sarcoma, the second most frequent bone cancer type, affects mainly adolescents, who have a survival of 50% 5 yr after diagnosis. Current treatments include a combination of surgery, radiotherapy and chemotherapy, which present potential serious side effects. Melatonin, a natural molecule without relevant side effects, has been previously shown to induce cytotoxicity in SK-N-MC cells, a Ewing sarcoma cell line. Here, we found that there is a synergy in the antitumor effect when melatonin (50 mum-1 mm) is combined with vincristine at the concentration of 5-10 nm or with ifosfamide at the range of 100 mum-1 mm. This synergism is due to the potentiation of cell death, particularly to the potentiation of apoptosis, i.e., mainly the extrinsic apoptotic pathway. There is a significant increase in the activation of caspase-3, -8, -9 and Bid when melatonin is combined with vincristine or ifosfamide compared to the individual treatments. Finally, there is also a potentiation of the early free radical production, likely dependent on the extrinsic apoptosis pathway activation, when the drugs are combined with melatonin. Other proteins which are related to this pathway including mitogen-activated protein kinase or protein kinase B/Akt are not involved in apoptosis induced by these agents separately or when combined. The results shown here together with the facts that: (i) no relevant side effects have been reported for melatonin and (ii) melatonin has a cytoprotective effect on noncancer cells, opens the door for a new approach in the treatment of the Ewing sarcoma family of tumors. PMID- 20025644 TI - Longitudinal assessment of fibrinogen in relation to subclinical cardiovascular disease: the CARDIA study. AB - OBJECTIVE: To examine the strength of the associations of fibrinogen with subclinical atherosclerosis in healthy persons. METHODS: A population-based, prospective, observational study of black and white men and women (Coronary Artery Risk Development in Young Adults [CARDIA]). Fibrinogen levels were measured at year 7 (ages 25-37, n = 2969), and again at year 20 (ages 38-50, n = 2832). Measures of subclinical atherosclerosis (coronary artery calcification [CAC] and carotid intimal-medial thickness [CIMT]) were recorded at year 20. RESULTS: Over the 13-year study interval (1992-1993 to 2005-2006), fibrinogen rose from a mean of 3.32 to 4.05 g L(-1). After adjusting for age, gender and race, fibrinogen was positively associated with greater incidence of CAC and increased CIMT cross-sectionally as well as after 13 years of follow-up (all P trend < 0.001). After further adjustment for field center, BMI, smoking, education, systolic blood pressure, diabetes, antihypertensive medication use, total and HDL cholesterol, and CRP, significant positive relationships between fibrinogen and incidence of CAC remained for the total cohort longitudinally (P trend = 0.037), but not cross-sectionally (P-trend = 0.147). CONCLUSION: This 13 year study demonstrates that higher levels of fibrinogen during young adulthood are positively associated with incidence of CAC and increased CIMT in middle-age, but the strength of the association declines with increasing age. PMID- 20025645 TI - Quality of life after pulmonary embolism: validation of the PEmb-QoL Questionnaire. AB - BACKGROUND: Even though quality of life (QoL) has become a key component of medical care, there is no instrument available that specifically measures QoL after pulmonary embolism (PE). Recently, the Pulmonary Embolism Quality of Life (PEmb-QoL) Questionnaire has been developed to address this gap. OBJECTIVE: To evaluate the validity of the PEmb-QoL questionnaire. METHODS: We distributed the PEmb-QoL questionnaire and the Short Form-36 (SF-36) questionnaire twice among consecutive subjects with a history of objectively confirmed acute PE. Internal consistency reliability, test-retest reliability, convergent validity and criterion validity, and correlations between the PEmb-QoL and clinical patient characteristics were assessed using standard-scale construction techniques. RESULTS: Ninety participants completed the questionnaires twice. Internal consistency was adequate (Cronbach's alpha 0.62-0.94), as well as test-retest reliability (intra-class correlation coefficients: 0.78-0.94). Furthermore, correlation between the PEmb-QoL questionnaire and the SF-36 questionnaire supported convergent validity. Age, obesity, cardiopulmonary comorbidity, centrally located PE and a family history of venous thromboembolism were shown to be independent determinants of disease-specific QoL. CONCLUSION: The PEmb-QoL questionnaire is a reliable instrument to specifically assess QoL following PE, which is helpful in the identification of patients with decreased QoL following acute PE. PMID- 20025646 TI - Pulmonary Embolism Severity Index and troponin testing for the selection of low risk patients with acute symptomatic pulmonary embolism. AB - BACKGROUND: The combination of the Pulmonary Embolism Severity Index (PESI) and troponin testing could help physicians identify appropriate patients with acute pulmonary embolism (PE) for early hospital discharge. METHODS: This prospective cohort study included a total of 567 patients from a single center registry with objectively confirmed acute symptomatic PE. On the basis of the PESI, each patient was classified into one of five classes (I-V). At the time of hospital admission, patients had troponin I (cTnI) levels measured. The endpoint of the study was all-cause mortality within 30 days after diagnosis. We calculated the mortality rates in four patient groups: group 1, PESI class I-II plus cTnI < 0.1 ng mL(-1); group 2, PESI classes III-V plus cTnI < 0.1 ng mL(-1); group 3, PESI classes I-II plus cTnI > or = 0.1 ng mL(-1); and group 4, PESI classes III-V plus cTnI > or = 0.1 ng mL(-1). RESULTS: The study cohort had a 30-day mortality of 10% [95% confidence interval (CI), 7.6-12.5%]. Mortality rates in the four groups were 1.3%, 14.2%, 0% and 15.4%, respectively. Compared with non-elevated cTnl, the low-risk PESI had a higher negative predictive value (NPV) (98.9% vs. 90.8%) and negative likelihood ratio (NLR) (0.1 vs. 0.9) for predicting mortality. The addition of non-elevated cTnI to low-risk PESI did not improve the NPV or the NLR compared with either test alone. CONCLUSIONS: Compared with cTnl testing, PESI classification more accurately identified patients with PE who are at low risk of all-cause death within 30 days of presentation. PMID- 20025647 TI - Evaluation of mutagenic and antimutagenic properties of some bioactive xanthone derivatives using Vibrio harveyi test. AB - AIMS: Drug safety evaluation plays an important role in the early phase of drug development, especially in the preclinical identification of compounds' biological activity. The Vibrio harveyi assay was used to assess mutagenic and antimutagenic activity of some aminoalkanolic derivatives of xanthone (1-5), which were synthesized and evaluated for their anticonvulsant and hemodynamic activities. METHODS AND RESULTS: A novel V. harveyi assay was used to assess mutagenic and antimutagenic activity of derivatives of xanthone 1-5. Two V. harveyi strains were used: BB7 (natural isolate) and BB7M (BB7 derivative containing mucA and mucB genes on a plasmid pAB91273, products of these genes enhance error-prone DNA repair). According to the results obtained, the most beneficial mutagenic and antimutagenic profiles were observed for compounds 2 and 3. A modification of the chemical structure of compound 2 by the replacement of the hydroxy group by a chloride improved considerably the antimutagenic activity of the compound. Thus, antimutagenic potency reached a maximum with the presence of tertiary amine and chloride atom in the side chain. CONCLUSIONS: Among the newly synthesized aminoalkanolic derivatives of xanthone with potential anticonvulsant properties, there are some compounds exhibiting in vitro antimutagenic activity. In addition, it appears that the V. harveyi assay can be applied for primary mutagenicity and antimutagenicity assessment of compounds. SIGNIFICANCE AND IMPACT OF THE STUDY: The obtained preliminary mutagenicity and antimutagenicity results encourage further search in the group of amino derivatives of xanthone as the potential antiepileptic drugs also presenting some antimutagenic potential. Furthermore, V. harveyi test may be a useful tool for compounds safety evaluation. PMID- 20025648 TI - Subinhibitory concentrations of licochalcone A decrease alpha-toxin production in both methicillin-sensitive and methicillin-resistant Staphylococcus aureus isolates. AB - AIM: To evaluate the effect of subinhibitory concentrations of licochalcone A (LicA) on alpha-toxin secretion in Staphylococcus aureus. METHODS AND RESULTS: A haemolysin assay was used to investigate the haemolytic activities in culture supernatants of both methicillin-sensitive and methicillin-resistant Staph. aureus isolates cultured with graded subinhibitory concentrations of LicA. Alpha toxin secretion was detected by immunoblot analysis. Moreover, quantitative RT PCR was performed to assess the influence of LicA on the transcription of hla (the gene encoding alpha-toxin) and agr (accessory gene regulator). Growth in the presence of LicA markedly inhibited the mRNA levels of hla and agr in Staph. aureus, resulting in a reduction of alpha-toxin secretion and, thus, haemolytic activities. CONCLUSION: The secretion of alpha-toxin in Staph. aureus is decreased by LicA; this effect may be partially dependent upon inhibition of the Agr two-component system. SIGNIFICANCE AND IMPACT OF THE STUDY: The findings in our study may support the use of LicA as a lead compound in the design of more potent antibacterial agents that are based on the chalcone template. PMID- 20025649 TI - Inactivation of Cronobacter spp. (Enterobacter sakazakii) in infant formula using lactic acid, copper sulfate and monolaurin. AB - AIMS: To investigate the effect of lactic acid (LA), copper (II), and monolaurin as natural antimicrobials against Cronobacter in infant formula. METHODS AND RESULTS: The effect of LA (0.1, 0.2 and 0.3% v/v), copper (II) (10, 50 and 100 microg ml(-1)) and monolaurin (1000, 2000, and 3000 microg ml(-1)) suspended into tween-80 or dissolved in ethanol against Cronobacter in infant formula was investigated. Reconstituted infant formula and powdered infant formula were inoculated with five strains of Cronobacter spp. at the levels of c. 1 x 10(6) CFU ml(-1) and 1 x 10(3) CFU g(-1), respectively. LA at 0.2% v/v had a bacteriostatic effect on Cronobacter growth, whereas 0.3% v/v LA resulted in c. 3 log(10) reduction. Copper (II) at the levels of 50 microg ml(-1) and 100 microg ml(-1) elicited c. 1 and 2 log(10) reductions, respectively. The combination of 0.2% LA and 50 microg ml(-1) copper (II) resulted in a complete elimination of the organism. Monolaurin exhibited a slight inhibitory activity against Cronobacter (c. 1.5 log(10) difference) compared to the control when ethanol was used to deliver monolaurin. CONCLUSIONS: A complete elimination of Cronobacter was obtained when a combination of sublethal concentrations of LA (0.2%) and copper (II) (50 microg ml(-1)) was used. SIGNIFICANCE AND IMPACT OF THE STUDY: The use of the synergistic interactive combination of LA and copper (II) could be beneficial to control Cronobacter in the infant formula industry. PMID- 20025650 TI - Molecular detection of the entomopathogenic bacterium Pseudomonas entomophila using PCR. AB - AIMS: To develop a specific, fast and simple molecular method useful to detect the entomopathogenic bacterium Pseudomonas entomophila. METHODS AND RESULTS: The use of bioinformatics tools allowed the identification of unique genes present in P. entomophila genome. Using such genes, we designed primers aiming to detect specifically P. entomophila by PCR. Furthermore, a pair of primers specifically designed to amplify the 16S rRNA gene in Pseudomonas species was used. Primer specificity was checked using environmental pseudomonad and nonpseudomonad species. A 618 -bp fragment was amplified only in Pseudomonas using the 16S rDNA primers. Primers (PSEEN1497) designed to detect P. entomophila amplified a 570 bp fragment only in P. entomophila. A duplex PCR was developed combining 16S rDNA and PSEEN1497 primers that allowed the detection of P. entomophila present in experimentally infected Drosophila melanogaster. CONCLUSIONS: We developed a molecular method useful to detect P. entomophila present in bacterial cultures or directly from infected insects. SIGNIFICANCE OF THE STUDY: To the best of our knowledge, this is the first molecular method aiming to detect P. entomophila in environmental samples. The use of our method will facilitate studies related to ecology and insect host range of this entomopathogenic bacterium. PMID- 20025652 TI - Genetic structure is correlated with phenotypic divergence rather than geographic isolation in the highly polymorphic strawberry poison-dart frog. AB - Phenotypic and genetic divergence can be influenced by a variety of factors, including sexual and natural selection, genetic drift and geographic isolation. Investigating the roles of these factors in natural systems can provide insight into the relative influences of allopatric and ecological modes of biological diversification in nature. The strawberry poison frog, Dendrobates pumilio, presents an excellent opportunity for this kind of research, displaying a diverse array of colour morphs and inhabiting a heterogeneous landscape that includes oceanic islands, fragmented rainforest patches and wide expanses of suitable habitat. In this study, we use 15 highly polymorphic microsatellite loci to estimate population structure and gene flow among populations from across the range of D. pumilio and a causal modelling framework to statistically test 12 hypotheses regarding the geographic and phenotypic variables that explain genetic differentiation within this system. Our results demonstrate that the genetic distance between populations is most strongly associated with differences in dorsal coloration. Previous experimental studies have shown that phenotypic differences can result in sexual and natural selection against non-native phenotypes, and our results now show that these forces lead to genetic isolation between different colour morphs in the wild, presenting a potential case of incipient speciation through selection. PMID- 20025653 TI - Low genetic diversity in a snail intermediate host (Biomphalaria pfeifferi Krass, 1848) and schistosomiasis transmission in the Senegal River Basin. AB - Population genetic perturbations of intermediate hosts, often a consequence of human pressure on environmental resources, can precipitate unexpectedly severe disease outbreaks. Such disturbances are set to become increasingly common following range changes concomitant with climate shifts, dwindling natural resources and major infrastructure changes such as hydroprojects. Construction of the Diama dam in the Senegal River Basin (SRB) reduced river salinity, enabling the freshwater snail intermediate host Biomphalaria pfeifferi to rapidly expand its distribution. A serious public health problem ensued, with an epidemic of intestinal schistosomiasis occurring in the previously schistosome-free Richard Toll region within 2 years. The current study aimed to assess the population variability of B. pfeifferi in the SRB, and speculate upon its subsequent impact on host-parasite interactions following such engineered ecological change. Genetic variation at nine polymorphic microsatellite loci revealed little population differentiation in SRB snails compared with those from natural habitats in Zimbabwe, where Schistosoma mansoni transmission is much lower. 'Open' SRB habitats are associated with greater water contact, smaller population sizes and less genetic diversity, with sites downstream of Richard-Toll showing greater inter- and intrapopulation variation, concomitant with less frequent human contact. These observations may be explained by rapid expansion into pristine habitat selecting for high fecundity genotypes at the expense of schistosome resistance, presenting S. mansoni with genetically homogenous highly fecund susceptible populations around the focal point, promoting development of a highly compatible host-parasite relationship. Longitudinal study of such systems may prove important in predicting public health risks engendered by future environmental engineering projects. PMID- 20025654 TI - Genetic isolation among sympatric vegetative compatibility groups of the aflatoxin-producing fungus Aspergillus flavus. AB - Aspergillus flavus, a fungal pathogen of animals and both wild and economically important plants, is most recognized for producing aflatoxin, a cancer-causing secondary metabolite that contaminates food and animal feed globally. Aspergillus flavus has two self/nonself recognition systems, a sexual compatibility system and a vegetative incompatibility system, and both play a role in directing gene flow in populations. Aspergillus flavus reproduces clonally in wild and agricultural settings, but whether a cryptic sexual stage exists in nature is currently unknown. We investigated the distribution of genetic variation in 243 samples collected over 4 years from three common vegetative compatibility groups (VCGs) in Arizona and Texas from cotton using 24 microsatellite loci and the mating type locus (MAT) to assess population structure and potential gene flow among A. flavus VCGs in sympatric populations. All isolates within a VCG had the same mating type with OD02 having MAT1-2 and both CG136 and MR17 having MAT1-1. Our results support the hypothesis that these three A. flavus VCGs are genetically isolated. We found high levels of genetic differentiation and no evidence of gene flow between VCGs, including VCGs of opposite mating-type. Our results suggest that these VCGs diverged before domestication of agricultural hosts (>10,000 yr bp). PMID- 20025655 TI - Are Antarctic minke whales unusually abundant because of 20th century whaling? AB - Severe declines in megafauna worldwide illuminate the role of top predators in ecosystem structure. In the Antarctic, the Krill Surplus Hypothesis posits that the killing of more than 2 million large whales led to competitive release for smaller krill-eating species like the Antarctic minke whale. If true, the current size of the Antarctic minke whale population may be unusually high as an indirect result of whaling. Here, we estimate the long-term population size of the Antarctic minke whale prior to whaling by sequencing 11 nuclear genetic markers from 52 modern samples purchased in Japanese meat markets. We use coalescent simulations to explore the potential influence of population substructure and find that even though our samples are drawn from a limited geographic area, our estimate reflects ocean-wide genetic diversity. Using Bayesian estimates of the mutation rate and coalescent-based analyses of genetic diversity across loci, we calculate the long-term population size of the Antarctic minke whale to be 670,000 individuals (95% confidence interval: 374,000-1,150,000). Our estimate of long-term abundance is similar to, or greater than, contemporary abundance estimates, suggesting that managing Antarctic ecosystems under the assumption that Antarctic minke whales are unusually abundant is not warranted. PMID- 20025656 TI - The impact of epistatic selection on the genomic traces of selection. AB - The rapid accumulation of genomic data has led to an explosion of studies searching for signals of past selection left within DNA sequences. Yet the majority of theoretical studies investigating the traces of selection have assumed a simple form of selection, without interactions among selectively fixed sites. Fitness interactions-'epistasis'-are commonplace, however, and take on a myriad of forms (Whitlock et al. 1995; Segreet al. 2005; Phillips 2008). It is thus important to determine how such epistasis would influence selective sweeps. On p. 5018 of this issue, Takahasi (2009) explores the effect of epistasis on genetic variation neighbouring two sites that interact in determining fitness, finding that such epistasis has a dramatic impact on the genetic variability in regions surrounding the interacting sites. PMID- 20025657 TI - Host specificity, phenotype matching and the evolution of reproductive isolation in a coevolved plant-pollinator mutualism. AB - Coevolutionary interactions between plants and their associated pollinators and seed dispersers are thought to have promoted the diversification of flowering plants (Raven 1977; Regal 1977; Stebbins 1981). The actual mechanisms by which pollinators could drive species diversification in plants are not fully understood. However, it is thought that pollinator host specialization can influence the evolution of reproductive isolation among plant populations because the pollinator's choice of host is what determines patterns of gene flow in its host plant, and host choice may also have important consequences on pollinator and host fitness (Grant 1949; Bawa 1992). In this issue of Molecular Ecology, Smith et al. (2009) present a very interesting study that addresses how host specialization affects pollinator fitness and patterns of gene flow in a plant host. Several aspects of this study match elements of a seminal mathematical model of plant-pollinator codivergence (Kiester et al. 1984) suggesting that reciprocal selection for matched plant and pollinator reproductive traits may lead to speciation in the host and its pollinator when there is strong host specialization and a pattern of geographic subdivision. Smith et al.'s study represents an important step to fill the gap in our understanding of how reciprocal selection may lead to speciation in coevolved plant-pollinator mutualisms. PMID- 20025658 TI - Conformational flexibility of sigma(70) in anti-terminator loading. AB - In promoter DNA, the preferred distance of the -10 and -35 elements for interacting with RNA polymerase-bound sigma(70) is 17 bp. However, the Devi et al. paper in this issue of Molecular Microbiology demonstrates that when the C terminal domain of sigma(70), including the 3.2 linker, is not attached to the core enzyme, distances between 0 and 3 bp can be accommodated. This attests to the great flexibility of the 3.2 linker. The particularly stable complex with the 2 bp separation may lend itself to structural studies of an early elongation complex containing sigma(70). PMID- 20025660 TI - Some reassembly required. AB - Cells invest a significant amount of their energy synthesizing proteins, and a large portion of the energy expenditure goes into making ribosomes, the RNA protein machines at the centre of translation. When ribosomes are damaged in a cell, i.e. during stressful conditions, cells must first recognize the damage and then mount a response. Remme et al. show that instead of having to rebuild ribosomes from scratch, bacteria can repair ribosomes by replacing damaged proteins in situ, thereby saving significant time and energy. Given the central role of translation, such repair mechanisms might be widespread in nature. PMID- 20025659 TI - Two CDC42 paralogues modulate Cryptococcus neoformans thermotolerance and morphogenesis under host physiological conditions. AB - The precise regulation of morphogenesis is a key mechanism by which cells respond to a variety of stresses, including those encountered by microbial pathogens in the host. The polarity protein Cdc42 regulates cellular morphogenesis throughout eukaryotes, and we explore the role of Cdc42 proteins in the host survival of the human fungal pathogen Cryptococcus neoformans. Uniquely, C. neoformans has two functional Cdc42 paralogues, Cdc42 and Cdc420. Here we investigate the contribution of each paralogue to resistance to host stress. In contrast to non pathogenic model organisms, C. neoformans Cdc42 proteins are not required for viability under non-stress conditions but are required for resistance to high temperature. The paralogues play differential roles in actin and septin organization and act downstream of C. neoformans Ras1 to regulate its morphogenesis sub-pathway, but not its effects on mating. Cdc42, and not Cdc420, is upregulated in response to temperature stress and is required for virulence in a murine model of cryptococcosis. The C. neoformans Cdc42 proteins likely perform complementary functions with other Rho-like GTPases to control cell polarity, septin organization and hyphal transitions that allow survival in the environment and in the host. PMID- 20025661 TI - Overlapping repressor binding sites regulate expression of the Methanococcus maripaludis glnK(1) operon. AB - The euryarchaeal transcriptional repressor NrpR regulates a variety of nitrogen assimilation genes by 2-oxoglutarate-reversible binding to conserved palindromic operators. The number and positioning of these operators varies among promoter regions of regulated genes, suggesting NrpR can bind in different patterns. Particularly intriguing is the contrast between the nif and glnK(1) promoter regions of Methanococcus maripaludis, where two operators are present but with different configurations. Here we study NrpR binding and regulation at the glnK(1) promoter, where the two operator sequences overlap and occur on opposite faces of the double helix. We find that both operators function in binding, with a dimer of NrpR binding simultaneously to each overlapping operator. We show in vivo that the first operator plays a primary role in regulation and the second operator plays an enhancing role. This is the first demonstration of overlapping operators functioning in Archaea. PMID- 20025662 TI - Borrelia burgdorferi locus BB0795 encodes a BamA orthologue required for growth and efficient localization of outer membrane proteins. AB - The outer membrane (OM) of the pathogenic diderm spirochete, Borrelia burgdorferi, contains integral beta-barrel outer membrane proteins (OMPs) in addition to its numerous outer surface lipoproteins. Very few OMPs have been identified in B. burgdorferi, and the protein machinery required for OMP assembly and OM localization is currently unknown. Essential OM BamA proteins have recently been characterized in Gram-negative bacteria that are central components of an OM beta-barrel assembly machine and are required for proper localization and insertion of bacterial OMPs. In the present study, we characterized a putative B. burgdorferi BamA orthologue encoded by open reading frame bb0795. Structural model predictions and cellular localization data indicate that the B. burgdorferi BB0795 protein contains an N-terminal periplasmic domain and a C terminal, surface-exposed beta-barrel domain. Additionally, assays with an IPTG regulatable bb0795 mutant revealed that BB0795 is required for B. burgdorferi growth. Furthermore, depletion of BB0795 results in decreased amounts of detectable OMPs in the B. burgdorferi OM. Interestingly, a decrease in the levels of surface-exposed lipoproteins was also observed in the mutant OMs. Collectively, our structural, cellular localization and functional data are consistent with the characteristics of other BamA proteins, indicating that BB0795 is a B. burgdorferi BamA orthologue. PMID- 20025663 TI - Activation of the L,D-transpeptidation peptidoglycan cross-linking pathway by a metallo-D,D-carboxypeptidase in Enterococcus faecium. AB - Bypass of the penicillin-binding proteins by an L,D-transpeptidase (Ldt(fm)) confers cross-resistance to beta-lactam and glycopeptide antibiotics in mutants of Enterococcus faecium selected in vitro. Ldt(fm) is produced by the parental strain D344S although it insignificantly contributes to peptidoglycan cross linking as pentapeptide stems cannot be used as acyl donors by this enzyme. Here we show that production of the tetrapeptide substrate of Ldt(fm) is controlled by a two-component regulatory system (DdcRS) and a metallo-D,D-carboxypeptidase (DdcY). The locus was silent in D344S and its activation was due to amino acid substitutions in DdcS or DdcR that led to production of DdcY and hydrolysis of the C-terminal D-Ala residue of the cytoplasmic peptidoglycan precursor UDP MurNAc-pentapeptide. The T(161)A and T(161)M substitutions affected a position of DdcS known to be essential for the phosphatase activity of related sensor kinases. Complete elimination of UDP-MurNAc-pentapeptide, which was required specifically for resistance to glycopeptides, involved substitutions in DdcY that increased the catalytic efficiency of the enzyme (E(127)K) and affected its interaction with the cell envelope (I(14)N). The ddc locus displays striking similarities with portions of the van vancomycin resistance gene clusters, suggesting possible routes of emergence of cross-resistance to glycopeptides and beta-lactams in natural conditions. PMID- 20025664 TI - Deletion of dop in Mycobacterium smegmatis abolishes pupylation of protein substrates in vivo. AB - Proteasome-bearing bacteria make use of a ubiquitin-like modification pathway to target proteins for proteasomal turnover. In a process termed pupylation, proteasomal substrates are covalently modified with the small protein Pup that serves as a degradation signal. Pup is attached to substrate proteins by action of PafA. Prior to its attachment, Pup needs to undergo deamidation at its C terminal residue, converting glutamine to glutamate. This step is catalysed in vitro by Dop. In order to characterize Dop activity in vivo, we generated a dop deletion mutant in Mycobacterium smegmatis. In the Deltadop strain, pupylation is severely impaired and the steady-state levels of two known proteasomal substrates are drastically increased. Pupylation can be re-established by complementing the mutant with either DopWt or a Pup variant carrying a glutamate at its ultimate C terminal position (PupGGE). Our data show that Pup is deamidated by Dop in vivo and that likely Dop alone is responsible for this activity. Furthermore, we demonstrate that a putative N-terminal ATP-binding motif is crucial for catalysis, as a single point mutation (E10A) in this motif abolishes Dop activity both in vivo and in vitro. PMID- 20025665 TI - The ribonucleases J1 and J2 are essential for growth and have independent roles in mRNA decay in Streptococcus pyogenes. AB - The paralogous ribonucleases J1 and J2, recently identified in Bacillus subtilis, have both endoribonucleolytic and 5'-to-3' exoribonucleolytic activities and participate in degradation and regulatory processing of mRNA. RNases J1 and J2 have partially overlapping target specificities, but only RNase J1 is essential for B. subtilis growth. Because mRNA decay is important in regulation of virulence factors of Streptococcus pyogenes (the group A streptococcus, GAS), we investigated the role of these newly described RNases in GAS. We found that conditional mutants for both RNases J1 and J2 require induction for growth, so we conclude that, unlike the case in B. subtilis, both of these RNases are essential for GAS growth, and therefore their functions are not redundant. We compared decay of representatives of the two classes of messages we had previously identified: Class I, which decay rapidly in exponential and stationary phase of growth (hasA and gyrA), and Class II, which are stable in stationary phase and exhibit a biphasic decay curve in exponential phase (sagA and sda). We report that RNases J1 and J2 affect the rate of decay of Class I messages and the length of the first phase in decay of Class II messages. PMID- 20025666 TI - The role of configuration and coupling in autoregulatory gene circuits. AB - Autoregulatory gene circuits can be physically encoded within the genome in a number of different configurations. By physical encoding, we mean the orientation and relative proximity of the genes within the circuit. In this work, we quantified the behaviour of an inducible, negatively autoregulated gene circuit arranged in different transcriptional configurations using the tetRA circuit from Tn10 as our basis. Mathematical modelling predicted that circuits arranged in configurations where the expression of the transcription factor is decoupled from its target genes afforded more flexibility relative to configurations where expression is coupled. We found that these decoupled configurations reduced the concentration of transcription factor needed to regulate inducible expression from the circuit. As lower concentrations of transcription factor were required, these decoupled configurations could also be activated at much lower concentrations of the inducer. We experimentally validated these predictions in Escherichia coli by comparing the response of synthetic circuits based on the tetRA circuit arranged in different configurations. Collectively, these results provide one example of how the arrangement of a gene circuit within the genome can affect its behaviour. PMID- 20025667 TI - Identification and characterization of the aspartate chemosensory receptor of Campylobacter jejuni. AB - Campylobacter jejuni is a highly motile bacterium that responds via chemotaxis to environmental stimuli to migrate towards favourable conditions. Previous in silico analysis of the C. jejuni strain NCTC11168 genome sequence identified 10 open reading frames, tlp1-10, that encode putative chemosensory receptors. We describe the characterization of the role and specificity of the Tlp1 chemoreceptor (Cj1506c). In vitro and in vivo models were used to determine if Tlp1 had a role in host colonization. The tlp1(-) isogenic mutant was more adherent in cell culture, however, showed reduced colonization ability in chickens. Specific interactions between the purified sensory domain of Tlp1 and l aspartate were identified using an amino acid array and saturation transfer difference nuclear magnetic resonance spectroscopy. Chemotaxis assays showed differences between migration of wild-type C. jejuni cells and that of a tlp1(-) isogenic mutant, specifically towards aspartate. Furthermore, using yeast two hybrid and three-hybrid systems for analysis of protein-protein interactions, the cytoplasmic signalling domain of Tlp1 was found to preferentially interact with CheV, rather than the CheW homologue of the chemotaxis signalling pathway; this interaction was confirmed using immune precipitation assays. This is the first identification of an aspartate receptor in bacteria other than Escherichia coli and Salmonella enterica serovar Typhimurium. PMID- 20025668 TI - An integrated regulatory network including two positive feedback loops to modulate the activity of sigma(E) in mycobacteria. AB - sigma(E), one of the best characterized mycobacterial extracytoplasmic function sigma factors, is involved in virulence, surface stress response and modulation of the inflammatory response during infection. The regulation of its activity is very complex and involves transcriptional, translational and post-translational control. Post-translational regulation is controlled by RseA, an anti-sigma factor belonging to the zinc-associated anti-sigma factor family. In this issue of Molecular Microbiology, Barik et al. demonstrate that RseA is a redox-sensing protein that is able to bind sigma(E) only in reducing environment. Importantly, they describe a novel positive feedback loop responsible for sigma(E) release and activation following surface stress, due to ClpC1P2-dependent proteolytic degradation of RseA, depending on its phosphorylation by the eukaryotic-like Ser/Thr protein kinase PknB. PMID- 20025669 TI - RseA, the SigE specific anti-sigma factor of Mycobacterium tuberculosis, is inactivated by phosphorylation-dependent ClpC1P2 proteolysis. AB - Central to the response of Mycobacterium tuberculosis to environmental stress is the regulation of genes under the control of alternative sigma factors. Sigma E of M. tuberculosis plays an important role in the intracellular life of the bacterium and regulates several genes which are important for maintaining the integrity of the cell envelope stress. This makes it important to understand how SigE is activated under stress. Here we elucidate the mechanisms regulating interaction of SigE with its cognate anti-sigma factor RseA. Cysteines 70 and 73 are required for redox-dependent interaction of RseA with SigE. Under surface stress, PknB-dependent phosphorylation of RseA on T39 is required for its cleavage by ClpC1P2 thereby activating the SigE regulon. Rv2745c (MSMEG_2694), a transcriptional regulator, activates the clp regulon in response to vancomycin induced stress. Taken together with the previous report that Rv2745c is activated by SigE, our study uncovers a positive feedback loop that activates the sigE regulon under envelope stress. PMID- 20025670 TI - Direct MinE-membrane interaction contributes to the proper localization of MinDE in E. coli. AB - Dynamic oscillation of the Min system in Escherichia coli determines the placement of the division plane at the midcell. In addition to stimulating MinD ATPase activity, we report here that MinE can directly interact with the membrane and this interaction contributes to the proper MinDE localization and dynamics. The N-terminal domain of MinE is involved in direct contact between MinE and the membranes that may subsequently be stabilized by the C-terminal domain of MinE. In an in vitro system, MinE caused liposome deformation into membrane tubules, a property similar to that previously reported for MinD. We isolated a mutant MinE containing residue substitutions in R10, K11 and K12 that was fully capable of stimulating MinD ATPase activity, but was deficient in membrane binding. Importantly, this mutant was unable to support normal MinDE localization and oscillation, suggesting that direct MinE interaction with the membrane is critical for the dynamic behavior of the Min system. PMID- 20025671 TI - Transcription factor AP2-Sp and its target genes in malarial sporozoites. AB - The malarial sporozoite is the stage that infects the liver, and genes expressed in this stage are potential targets for vaccine development. Here, we demonstrate that specific gene expression in this stage is regulated by an AP2-related transcription factor, designated AP2-Sp (APETALA2 in sporozoites), that is expressed from the late oocyst to the salivary gland sporozoite. Disruption of the AP2-Sp gene did not affect parasite replication in the erythrocyte but resulted in loss of sporozoite formation. The electrophoretic mobility-shift assay showed that the DNA-binding domain of AP2-Sp recognizes specific eight-base sequences, beginning with TGCATG, which are present in the proximal promoter region of all known sporozoite-specific genes. Promoter assays demonstrated that these sequences act as cis-acting elements and are critical for the expression of sporozoite-specific genes with different expression profiles. In transgenic parasites that express endogenous AP2-O (APETALA2 in ookinetes), but whose AP2 domain had been swapped with that of AP2-Sp, several target genes of AP2-Sp were induced in the ookinete stage. These results indicate that AP2-Sp is a major transcription factor that regulates gene expression in the sporozoite stage. PMID- 20025672 TI - Bacillus subtilis ribonucleases J1 and J2 form a complex with altered enzyme behaviour. AB - Ribonucleases J1 and J2 are recently discovered enzymes with dual 5'-to-3' exoribonucleolytic/endoribonucleolytic activity that plays a key role in the maturation and degradation of Bacillus subtilis RNAs. RNase J1 is essential, while its paralogue RNase J2 is not. Up to now, it had generally been assumed that the two enzymes functioned independently. Here we present evidence that RNases J1 and J2 form a complex that is likely to be the predominant form of these enzymes in wild-type cells. While both RNase J1 and the RNase J1/J2 complex have robust 5'-to-3' exoribonuclease activity in vitro, RNase J2 has at least two orders of magnitude weaker exonuclease activity, providing a possible explanation for why RNase J1 is essential. The association of the two proteins also has an effect on the endoribonucleolytic properties of RNases J1 and J2. While the individual enzymes have similar endonucleolytic cleavage activities and specificities, as a complex they behave synergistically to alter cleavage site preference and to increase cleavage efficiency at specific sites. These observations dramatically change our perception of how these ribonucleases function and provide an interesting example of enzyme subfunctionalization after gene duplication. PMID- 20025673 TI - A mutational analysis reveals new functional interactions between domains of the Oxa1 protein in Saccharomyces cerevisiae. AB - The Oxa1/YidC/Alb3 family plays a key role in the biogenesis of the respiratory and photosynthetic complexes in bacteria and organelles. In Saccharomyces cerevisiae, Oxa1 mediates the co-translational insertion of mitochondrially encoded subunits of the three respiratory complexes III, IV and V within the inner membrane and also controls a late step in complex V assembly. No crystal structure of YidC or Oxa1 is available and little is known about the respective role of each transmembrane segment (TM) and hydrophilic loop of this polytopic protein on the biogenesis of the three complexes. Here, we have generated a collection of random point mutations located in the hydrophobic and hydrophilic domains of the protein and characterized their effects on the assembly of the three respiratory complexes. Our results show mutant-dependent differential effects, particularly on complex V. In order to identify tertiary interactions within Oxa1, we have also isolated revertants carrying second-site compensatory mutations able to restore respiration. This analysis reveals the existence of functional interactions between TM2 and TM5, TM4 and TM5 as well as between TM4 and loop 2, highlighting the key position of TM4 and TM5 in the Oxa1 protein. PMID- 20025674 TI - Both genes in the Marinomonas mediterranea lodAB operon are required for the expression of the antimicrobial protein lysine oxidase. AB - The melanogenic marine bacterium Marinomonas mediterranea synthesizes a novel antimicrobial protein (LodA) with lysine-epsilon oxidase activity (EC 1.4.3.20). Homologues to LodA have been detected in several Gram-negative bacteria, where they are involved in biofilm development. Adjacent to lodA is located a second gene, lodB, of unknown function. This genomic organization is maintained in all the microorganisms containing homologues to these genes. In this work we show that lodA and lodB constitute an operon. Western blot analysis and enzymatic determinations revealed that LodA is secreted to the external medium when the culture reaches the stationary phase. LodB, on the other hand, has only been detected inside cells, but it is not secreted. The expression of the lysine epsilon oxidase (LOD) activity in M. mediterranea requires functional copies of both genes since mutants lacking either lodA or lodB do not show any LOD activity. The active form of LodA containing the quinonic cofactor is intracellularly generated in a process that takes place only in the presence of LodB, suggesting that the latter is involved in this process. Moreover, in the absence of one of the proteins, the stability of the partner protein is compromised leading to a marked decrease in its cellular levels. PMID- 20025675 TI - Performance characteristics of scintigraphic colon transit measurement in health and irritable bowel syndrome and relationship to bowel functions. AB - BACKGROUND: The inter- and intra-subject variations of scintigraphy, which are used to identify colonic transit disturbances in irritable bowel syndrome (IBS), are unclear. The relationship between colonic transit and bowel functions is incompletely understood. To assess inter- and intra-subject variations of scintigraphic colonic transit measurements in 86 IBS patients and 17 healthy subjects and to quantify the relationship between colonic transit and bowel symptoms in 147 IBS patients and 46 healthy subjects. METHODS: Data from participants with multiple colonic transit measurements were analysed. Primary end points were colonic filling at 6 h (CF6h) and geometric center (GC) at 24 and 48 h for colonic transit. Bowel functions were assessed by daily stool diaries. KEY RESULTS: Inter- and intra-subject variations were greater for small intestinal than colonic transit. Overall, inter- and intra-subject variations were relatively narrow for colonic transit (both GC24h and GC48h, with lower COV at 48 h); there was little intra-subject variation in health and IBS-constipation over a period of T, c.-670C>T) and SNP-5 (rs3738423:C>T, c.288C>T) were selected as tagging single nucleotide polymorphisms (SNP) and haplotypes were reconstructed. Association was analyzed in MCD patients. Then, the identified SNP site was analyzed in IgAN patients with mild histological changes (Haas subclass I and II). RESULTS: The C allele and CC genotype frequencies at the SNP-2 site, as well as the frequency of haplotype CC, were significantly lower in MCD patients than in healthy controls. Furthermore, they were also associated with the degree of proteinuria in MCD patients. But in IgAN patients, no such association was identified. CONCLUSION: The study suggested the polymorphism and haplotype of NPHS2 gene were associated with the genetic susceptibility and also the degree of proteinuria to MCD. Proteinuria in MCD and IgAN might occur through different mechanisms. PMID- 20025682 TI - Potential for erythropoietin synthesis in kidney of uraemic rat alters depending on severity of renal failure. AB - AIM: Renal anaemia is a common early complication of chronic renal failure (CRF) that is characterized by relative erythropoietin (EPO) deficiency. Although a lowered renal function is considered to induce limited EPO production, potential EPO production capacity in CRF remains unclear. The aim of this study was to determine the mechanisms underlying this relative deficiency. METHODS: Male Sprague-Dawley rats were underwent 5/6 nephrectomy with different severities of CRF. These rats were assigned to two groups - mild CRF or advanced CRF - and subjected to haemodilution by exchange of blood with Ringer's solution or haemoconcentration by blood transfusion. Serum EPO and EPO transcript levels in remnant kidney were examined. Expression levels of hypoxia-related genes, including heme oxygenase-1 (HO-1) and glucose transporter-1 (Glut-1), were also examined. RESULTS: Haemodilution increased both serum EPO and EPO transcript levels in mild CRF, as observed in sham-operated controls, whereas the extents of such increases were significantly smaller in advanced CRF. HO-1 and Glut-1 transcript levels also increased by haemodilution in mild CRF, but not in advanced CRF. Haemoconcentration markedly decreased serum EPO and EPO transcript levels in mild CRF as in controls. Rats with advanced CRF did not survive after blood transfusion. CONCLUSION: Potential EPO regulation capacity in mild CRF is as conserved as that in normal control, whereas that in advanced CRF is impaired, suggesting that underlying mechanisms of low EPO production alters according to the stage of CRF. PMID- 20025683 TI - Chronic kidney disease in Tasmania. AB - AIM: Chronic kidney disease (CKD) is a progressive disease which is becoming a major public health issue due to its high rate of premature death, poor quality of life and expensive end-stage treatment (dialysis or transplantation). The burden of this chronic condition in a community setting was examined. METHODS: Data were obtained from 369,098 Tasmanian adults (aged >or=18 years) and included 1,640,687 measurements of creatinine taken between 1995 and 2007. In 2007 alone, testing comprised 25.5% of the state's adult population. A modelled estimate of CKD prevalence was developed. RESULTS: For those at risk of CKD (aged >50 years), 50.6%, 70.2% and 82% had a measured creatinine (and reported estimated glomerular filtration rate (eGFR)) during the last 1, 2 and 3 years respectively. However, only 9.4% of people with eGFR of less than 60 mL/min per 1.73 m(2) had albuminuria formally measured. Estimated prevalence of stage III or greater CKD (eGFR <60 mL/min per 1.73 m(2)) was at least 11.4% of women and 8.6% of men during 2007. Detection of low eGFR increased significantly over the last 13 years. There was a large geographic variation throughout Tasmania and high relative mortality with lower eGFR. There is a broad gap between the number of people with eGFR of less than 15 mL/min per 1.73 m(2) (stage V CKD) and those receiving dialysis treatment. CONCLUSION: The number of people identified with low eGFR has increased significantly since 1995 with a large geographic variation. Despite this, testing for kidney disease (by measuring serum creatinine and albuminuria) in people at risk is still suboptimal. PMID- 20025684 TI - Influence of ketoanalogs supplementation on the progression in chronic kidney disease patients who had training on low-protein diet. AB - AIM: A low-protein diet (LPD) is a conservative treatment in patients with chronic kidney disease (CKD) to improve uremic symptoms and slow the progression of renal dysfunction. However, the deleterious effects of protein restriction on nutritional status have raised concern. We investigated whether ketoanalogs supplementation in CKD patients who had training on LPD retards the progression of CKD and maintains nutritional status. METHODS: Data were collected retrospectively from 120 consecutive patients in the CKD stages III and IV. Firstly all patients were restricted to LPD alone for 6 months (LPD alone), and then ketoanalogs of essential amino acids (KA) were supplemented for 6 months. RESULTS: The adequate LPD had not achieved in both periods. The declining slopes of glomerular filtration rate (GFR) during the LPD + KA period were significantly lower than those during the LPD alone period. This improvement in GFR was apparent in both subjects with diabetics and non-diabetic patients. Mean serum total cholesterol levels decreased in LPD + KA compared with LPD alone period. However, serum albumin levels did not change. Responders showed a higher prevalence of diabetes and higher serum albumin levels during the LPD alone period. Multivariate analysis revealed that responsiveness to LPD + KA was independently related to diabetes (p = 0.006) and high serum albumin levels (p = 0.011) in the LPD alone period. CONCLUSION: KA supplementation on over LPD delayed the progression of CKD without deteriorating nutritional status, and initial serum albumin levels could be an independent factor. PMID- 20025685 TI - Long-term graft function with tacrolimus and cyclosporine in renal transplantation: paired kidney analysis. AB - AIM: The first prospective, randomized trial with paired kidney analysis was conducted to compare the efficacy and safety of tacrolimus with cyclosporine based immunosuppressive therapy in renal transplant recipients. This paper reports the long-term follow-up results of the authors' previously published study, with the main focus on graft survival and renal function. METHODS: Chinese patients transplanted in our centre between June 1998 and June 2005 with their first deceased renal transplant were included. Patients were included if both kidneys were received by the authors' centre, thus allowing a paired analysis. Patients were randomized to receive triple immunosuppressive therapy with either tacrolimus or Neoral cyclosporine, concomitantly with prednisolone and azathioprine therapy. RESULTS: Seventy-six patients received cadaveric kidneys from 38 donors. Each pair of kidneys was randomly assigned to a separate group (38 subjects/group). The mean follow-up duration was 6.1 +/- 1.8 years. The mean calculated creatinine clearance was significantly higher in patients receiving tacrolimus-based therapy. The rate of biopsy-proven acute rejection was lower in the tacrolimus group (18.4% vs 42.1%, P = 0.03). The patient and graft survival were comparable in both treatment arms. Significantly fewer patients on tacrolimus-based therapy developed hypercholesterolaemia (P = 0.05). However, there was no significant difference in the development of post-transplant diabetes mellitus, hypertension, opportunistic infection and malignancy between both groups. CONCLUSION: Using the immunosuppressive regimen, tacrolimus-based therapy provided adequate immunosuppression with better renal function and less acute rejection, as compared with cyclosporine-based therapy. PMID- 20025686 TI - Hepatocyte nuclear factor-1beta frame-shift mutation in sporadic glomerulocystic kidney disease associated with agenesis of the corpus callosum. PMID- 20025687 TI - Left ventricular false tendon in a dialysis patient. PMID- 20025688 TI - Defining deficiency of vitamin D based on 25(OH)D concentration: is it real? PMID- 20025692 TI - Lower functional gastrointestinal disorders: evidence of abnormal colonic transit in a 287 patient cohort. AB - BACKGROUND Abnormalities of colonic motility were reported in relatively small studies of patients with lower functional gastrointestinal disorders (FGID) including irritable bowel syndrome (IBS). The influence of gender and body mass on the observed motor pathophysiology is unclear. We sought to compare colonic transit in patients within different lower FGID subgroups and healthy controls, controlling for gender and BMI, and to determine whether BMI independently influences colonic motility. METHODS We evaluated a scintigraphic gastrointestinal and colonic transit database of 287 lower FGID patients associated with constipation (IBS-C, or functional constipation, n = 118), diarrhoea (IBS-D or functional diarrhoea, n = 139) or mixed bowel function (IBS M, n = 30) and 170 healthy controls. We measured colon filling at 6 h (CF 6 h), and overall colonic transit at 8, 24 and 48 h. KEY RESULTS Colon filling at 6 h did not differentiate health from FGID. Colonic transit was abnormal at 24 h (GC24 of <1.50 or >3.86) in 29.7% of all lower FGID patients. There was a significant overall association between colonic transit and subject group (healthy controls and FGID subgroups) at 8 (P = 0.01), 24 (P < 0.001) and 48 h (P < 0.001) in particular for those with diarrhoea or constipation at 24 and 48 h (P < 0.05), even after adjusting for age, gender and BMI. In addition, BMI was associated with colonic transit after adjusting for age, gender and subject group. CONCLUSIONS & INFERENCES Abnormal transit is documented non-invasively with scintigraphy in 30% of lower FGID patients; transit measurement may help document pathophysiology and inform selection of therapy in lower FGID. PMID- 20025693 TI - Long-term changes in blood pressure following orlistat and sibutramine treatment: a meta-analysis. AB - Previous meta-analyses investigating blood pressure effects of anti-obesity drugs have included studies using non-licensed doses, but not data from head-to-head studies. Furthermore, although diabetes is an important comorbidity in obesity, variation in blood pressure effects across diabetes status has not been investigated. The objective of this study was to estimate the effects on systolic (SBP) and diastolic blood pressure (DBP) of orlistat and sibutramine. Medline, EMBASE, the Cochrane controlled trials register and reference lists of identified articles from 1990 to February 2009 were searched. All placebo-controlled randomized controlled trials of 12-month duration or randomized head-to-head studies of any duration on adults using standard doses were included. Studies/study arms were excluded if they only evaluated weight maintenance after weight loss. Randomized controlled trials were identified, subjected to inclusion and exclusion criteria, and reviewed. Random effects models were used for assessment of weighted mean differences. Eighteen placebo-controlled (12 orlistat, 5540 patients; 6 sibutramine, 1495 patients) and four head-to-head trials (348 patients) met the inclusion criteria. Three orlistat and three sibutramine studies examined overweight subjects with type 2 diabetes (T2DM), as did two head-to-head trials. Mean baseline SBP ranged from 119 to 153 mmHg, and mean DBP from 69 to 98 mmHg. Overall, the placebo-controlled SBP change was -1.9 (95% CI; -2.7, -1.1) mmHg for orlistat, and 0.5 (-1.1, 2.1) mmHg for sibutramine. The corresponding values for DBP were -1.5 (-2.2, -0.8) and 1.7 (0.7, 2.6). Compared with patients without diabetes, diabetic patients treated with orlistat experienced smaller and non-significant reductions of SBP (-0.9; -2.6, 0.7 vs. 2.2; -3.0, -1.3) and DBP (-1.0; -2.4, 0.3 vs. -1.6; -2.4, -0.8). For sibutramine, higher on-treatment elevations in SBP (1.6; -1.3, 4.5 vs. 0.1; -1.8, 2.0) and DBP (2.4; 0.6, 4.1 vs. 1.4; 0.3, 2.5) were seen in patients with vs. without diabetes. In head-to-head trials, the overall differences between sibutramine and orlistat were small and non-significant for both SBP (1.0; -2.3, 4.3) and DBP ( 0.2; -2.9, 2.5). In conclusion, in the studies using approved sibutramine doses, the drug caused significant elevations in DBP, while the overall SBP effect was near null. Moreover, absence of a blood pressure-lowering effect of orlistat ad a higher DBP elevation by sibutramine were observed for persons with diabetes. Head to-head studies indicated that an indirect comparison of placebo-adjusted blood pressure effects may overestimate the adverse effects associated with sibutramine, but these studies were small, of shorter duration and of lower quality. PMID- 20025694 TI - Cholesterol imbalance in adipocytes: a possible mechanism of adipocytes dysfunction in obesity. AB - Studies of the past decade have increased our understanding of the role of adipose tissue dysfunction in obesity and obesity-related insulin resistance and type 2 diabetes. Although adipose tissue is the body's largest pool of free cholesterol, adipocytes have limited activity in cholesterol synthetic pathway. Thus, the majority of adipocyte cholesterol originates from circulating lipoproteins. To maintain cholesterol homeostasis, adipocytes have developed multiple pathways for cholesterol efflux. Several transcriptional factors, such as sterol regulatory element-binding proteins and liver X receptors may be responsible for the regulation of cholesterol homeostasis in adipocytes. Most notably, because altering cholesterol balance profoundly modifies adipocyte metabolism in a way resembling that seen in hypertrophied adipocytes, cholesterol imbalance is recognized as a characteristic for enlarged adipocytes per se in the obese state. In addition, plasma membrane cholesterol normalization by chromium picolinate can fully restore insulin-stimulated glucose transport, further supporting the role of the adipocyte cholesterol imbalance in obesity and insulin resistance. PMID- 20025695 TI - Obesity and post-operative complications in patients undergoing non-bariatric surgery. AB - As the prevalence of obesity continues to rise in society, an increasing number of patients undergoing non-bariatric surgery will be obese. Obesity is known to increase morbidity and mortality in the general population and thus is perceived as a risk factor for adverse post-surgical outcomes. This association is not clear-cut, however, and there is a lack of consensus in the literature on the risk between obesity and specific complications, in particular relating to infection, wound healing, respiratory and venous thromboembolism. The paucity of studies, as well as a lack of consistency of definition of obesity, with an over reliance on body mass index rather than body composition analysis, may underlie this confusion. Emerging concepts position central/visceral adipose tissue as potentially key to the pathogenesis of the comorbidities associated with obesity, thus this article reviews emerging research investigating the association between visceral obesity, the metabolic syndrome and resulting post-operative complications. It is hypothesized that the state of chronic inflammation and dysmetabolism observed in visceral obese patients negatively influences post operative outcomes and represents a potential target for pharmaconutrition. The need for further research investigating the influence of visceral adiposity on immune function post surgery and its impact on post-operative morbidity and mortality is highlighted. PMID- 20025696 TI - Trends in overweight and obesity in Swedish schoolchildren 1999-2005: has the epidemic reached a plateau? AB - The aim of this review is to synthesize published evidence on the most recent trends in overweight and obesity among Swedish children. Specifically, trends are reported among fourth graders (10-11 years) from six different municipalities between 1999 and 2005. Weights and heights in representative samples of children within each area were measured by school nurses as part of routine school health examinations. Standardized definitions of overweight, obesity and thinness were calculated by methods described by Cole et al. in 2000 and 2007. In Stockholm, obesity prevalence during academic years starting 1999 and 2003 decreased non significantly from 4.4% to 2.8% in girls, and increased non-significantly from 3.2% to 3.8% among boys. In Gothenburg, comparing academic years starting 2000 and 2004, prevalence of overweight in girls decreased from 19.6% to 15.9% (P < 0.01) while thinness increased from 9.5% to 11.9% (P < 0.05); no significant changes were observed in boys. Finally, the Swedish National Institute of Public Health released figures from Karlstad, Umea, Vasteras and Ystad in 2003-2005 during which time no trends in prevalence could be clearly shown. The stabilized rates are probably a result of regional and local actions that have taken place in many sectors of society, rather than one specific measure or national political action. PMID- 20025698 TI - Impact of ICD battery longevity on need for device replacements-insights from a Veterans Affairs database. AB - BACKGROUND: The number of implantable cardioverter defibrillator (ICD) replacements is increasing, which adds to patient risk and costs. OBJECTIVES: To understand the impact of increasing ICD longevity on need for replacements, costs, and some of the risks. METHODS: Using the Veterans Affairs records, veterans with ICD implants between June 1992 and April 2007 and dead as of April 2009 were identified. Data were obtained by retrospective records review. The longevity of each ICD was the duration from implant to explant. For each ICD, the longevity needed to avoid one replacement was defined as longevity of that ICD plus the longevity of the subsequent ICD. RESULTS: The study cohort had 164 patients with 301 ICD implants. Ninety-two patients had at least one ICD replacement. Two patients were excluded for missing data. Twenty-seven patients had an ICD explanted for reasons other than battery depletion. Sixty-three patients received 83 ICDs for battery depletion alone. Among 27 patients who had ICD replacements for other reasons, four patients may have avoided a device infection related to ICD replacement if the initial ICD had lasted 7 years. If all ICDs had lasted 5, 7, or 9 years, then 26%, 58%, and 84% of patients, respectively, would not have needed an ICD replacement. Also 17, 37, and 53 ICD replacements, respectively, would have been avoided, saving US$314,500-US$980,500 over 15 years at 2005 Medicare reimbursement rates. CONCLUSIONS: Prolongation of battery life to 7 or 9 years is important to reduce patient risks and decrease costs. PMID- 20025697 TI - Implantable cardioverter defibrillator therapy for prevention of sudden cardiac death in children in the Netherlands. AB - INTRODUCTION: Implantable cardioverter defibrillator (ICD) therapy is increasingly used in children. The purpose of this multicenter study is to evaluate mid-term clinical outcome and to identify predictors for device discharge in pediatric ICD recipients. METHODS AND RESULTS: From 1995 to 2006, 45 patients in The Netherlands under the age of 18 years received an ICD. Mean age at implantation was 10.8 +/- 5.2 years. Primary prevention (N = 22) and secondary prevention (N = 23) were equally distributed. Underlying cardiac disorders were primary electrical disease (55%), cardiomyopathy (20%), and congenital heart disease (17%). The follow-up was 44 +/- 32.9 months. Three patients (7%) died and one patient (2%) underwent heart transplantation. ICD-related complications occurred in eight patients (17%), seven of whom had lead-related complications. Fourteen patients (31%) received appropriate ICD shocks; 12 patients (27%) received inappropriate ICD shocks. Fifty-five percent of 22 ICD recipients under the age of 12 years received appropriate shocks, which was higher as compared with 9% of 23 older ICD recipients (P = 0.003). Although the incidence of appropriate shocks in the present study was larger in secondary prevention (9/23; 39%) as compared with primary prevention (5/22; 23%), this difference did not reach significance. CONCLUSIONS: In our population of patients, children <12 years of age had more appropriate shocks than patients 13-18 years. The complication rate is low, and is mainly lead related. PMID- 20025699 TI - Vacuum-assisted wound closure for pacemaker infection. AB - BACKGROUND: Although infection is a serious complication of pacemaker implantation, optimal treatment of infections related to pacemaker systems is poorly defined. METHODS: We describe a man in his 60s, an octogenarian, and two nonagenarian females who were treated for an infected permanent pacemaker. All of these patients developed inflammation that presented as local symptoms and purulent collection in the pockets of their implanted pacemakers. After fenestration of the pacemaker pockets, they were treated with vacuum-assisted wound closure (VAC). RESULTS: Infection was eradicated in all the patients without the need for aggressive surgery. The open wound was re-sutured without complete removal of the pacemaker system in two patients. After removing the infected generator from the other two patients, the open wounds healed with or without re-suture. The mean duration of VAC was 19.5 days. The postoperative course of all of the patients was uneventful, and they remained completely asymptomatic after VAC, with no evidence of recurrent infection for 5- 15 months after discharge. CONCLUSIONS: When the risk of total system explantation is high, less-invasive VAC might serve as the option for treating an infected pacemaker. (PACE 2010; 426-430). PMID- 20025700 TI - From defibrillation theory to clinical implications. AB - BACKGROUND: Our defibrillation theory claims that the mean voltage threshold is a hyperbolic function of pulse duration and that voltages below rheobase should be avoided as being counterproductive. Truncation of the pulse just at rheobase level yields minimal stored energy thresholds. To verify or falsify this theory, animal experiments were carried out. MATERIAL AND METHODS: In two animal experiments, 212 defibrillation thresholds in 22 swine were determined with different biphasic pulses of which 92 were optimally truncated in phase 1. Step up test procedure was used with the first successful shock defined as "threshold." RESULTS: Experimental proof is gained that truncation according to "rheobase condition" shows lowest stored energy. A ranking order of stored energy thresholds demonstrates that (1) lower output capacitances reduce needed energy, and (2) pulse durations shorter or longer than optimal increase needed energy. The voltage-pulse-content threshold is linearly correlated with pulse duration. CONCLUSIONS: Truncation above or below rheobase increases the stored energy threshold. Voltage averaged during pulse duration is a hyperbolic function of pulse duration. The stored energy is reduced with decreasing output capacitance. The experimental results do not only fully verify our theory, they also suggest clinical implications: (1) the current usage of the "constant tilt concept" in implantable cardioverter defibrillator (ICD) should be abandoned in favor of "optimal truncation concept," (2) an algorithm developed for calculating optimal truncation proved to be useful so that incorporation into ICD for automatic adjustment is recommended, and (3) the output capacitance should be reduced from about 100 microF to 60 to 70 microF. PMID- 20025701 TI - Apparent AV dissociation upon induction of a narrow complex tachycardia. What is the mechanism? PMID- 20025702 TI - Severe venous and lymphatic obstruction after single-chamber pacemaker implantation in a patient with chest radiation therapy. AB - A 73-year-old woman with a history of paroxysmal atrial fibrillation, sinus node dysfunction, bilateral breast cancer, and extensive chest radiation developed progressive edema, dyspnea, and recurrent pleural effusions soon after single chamber pacemaker implantation. Thoracentesis yielded a diagnosis of chylothorax, and progressive refractory anasarca developed. A computed tomography angiogram suggested obstruction of the superior vena cava and left subclavian vein despite outpatient therapeutic anticoagulation. Autopsy confirmed venous thrombosis, along with mediastinal fibrosis. The presumed etiology of the chylothorax and anasarca was obstruction of the atretic central venous structures following pacemaker implantation, critically impairing the already tenuous venous and lymphatic drainage. (PACE 2010; 520-524). PMID- 20025703 TI - Cardiac resynchronization therapy in non-left bundle branch block morphologies. AB - INTRODUCTION: In select patients with systolic heart failure, cardiac resynchronization therapy (CRT) has been shown to improve quality of life, exercise capacity, ejection fraction (EF), and survival. Little is known about the response to CRT in patients with right bundle branch block (RBBB) or non specific intraventricular conduction delay (IVCD) compared with traditionally studied patients with left bundle branch block (LBBB). METHODS: We assessed 542 consecutive patients presenting for the new implantation of a CRT device. Patients were placed into one of three groups based on the preimplantation electrocardiogram morphology: LBBB, RBBB, or IVCD. Patients with a narrow QRS or paced ventricular rhythm were excluded. The primary endpoint was long-term survival. Secondary endpoints were changes in EF, left ventricular end-diastolic and systolic diameter, mitral regurgitation, and New York Heart Association (NYHA) functional class. RESULTS: Three hundred and thirty-five patients met inclusion criteria of which 204 had LBBB, 38 RBBB, and 93 IVCD. There were 32 deaths in the LBBB group, 10 in the RBBB, and 27 in the IVCD group over a mean follow up of 3.4 +/- 1.2 years. In multivariate analysis, no mortality difference amongst the three groups was noted. Patients with LBBB had greater improvements in most echocardiographic endpoints and NYHA functional class than those with IVCD and RBBB. CONCLUSION: There is no difference in 3-year survival in patients undergoing CRT based on baseline native QRS morphology. Patients with RBBB and IVCD derive less reverse cardiac remodeling and symptomatic benefit from CRT compared with those with a native LBBB. PMID- 20025704 TI - Relationship between the duration of the basal QRS complex and electrical therapies for ventricular tachycardias among ICD patients. AB - BACKGROUND: In implantable cardioverter-defibrillators (ICD) patients, the duration of the basal QRS complex (QRSd) is not associated with a greater risk of developing ventricular tachyarrhythmias. QRSd could be inversely related to the effectiveness of antitachycardia pacing (ATP) because it may be associated with longer conduction times of the paced-impulses and hence, with a greater propensity to require shocks to terminate ventricular tachycardias (VTs). METHODS: We followed 216 ICD patients (pacing site: right ventricular apex; QRSd 100 remained as an independent predictor of receiving shocks to terminate VTs (P = 0.01). According to Kaplan-Meier analysis, the occurrence of VTs was similar regardless of the QRSd (30% vs 38%; P = 0.2), but the incidence of shock due to VTs was higher in patients with a QRSd > 100 (19% vs 7%; P = 0.01). CONCLUSION: Since QRSd is a negative and independent predictor of effective ATP, ICD patients with QRSd > 100 ms require shocks more frequently to terminate VTs. PMID- 20025705 TI - Variability of left ventricular electromechanical activation during right ventricular pacing: implications for the selection of the optimal pacing site. AB - BACKGROUND: The right ventricular septum (RVS) and Hisian area (HA) are considered more "physiological" pacing sites than right ventricular apex (RVA). Studies comparing RVS to RVA sites have produced controversial results. There are no data about variability of electromechanical activation obtained by an approach using fluoroscopy and electrophysiological markers. This study compared the variability of left ventricular (LV) electromechanical activation in patients undergoing short-term RVA and RVS with that measured during HA pacing based on fluoroscopy and electrophysiological markers. METHODS: Tissue Doppler echocardiography was performed in 142 patients before and after RVA (54), RVS (44), and HA (44) pacing. Electromechanical activation was assessed by: (1) electromechanical latency (EML)-interval between QRS onset and mechanical activation of basal LV; (2) intra-LV dyssynchrony (intra-LV)-interval between earliest to the latest LV basal motion. The intra- and interpatients variability among pacing groups were assessed. RESULTS: Pacing from RVA showed longer EML and higher degree of intra-LV than RVS and HA pacing. RVA and RVS showed a higher variability than HA pacing with regard to intrapatient changes of EML (RVA vs RVS, P = 0.4; RVS vs HA, P = 0.01, RVA vs HA, P = 0.0002) and intra-LV (RVA vs RVS, P = 0.2; RVS vs HA, P = 0.04; RVA vs HA, P = 0.005). Similar results were found in interpatients variability from paced-values. CONCLUSIONS: RVA and RVS pacing produce a variable effect on LV electromechanical activation that is significantly more pronounced than HA pacing. A pacing site such as HA selected by fluoroscopic and electrophysiological markers maintains baseline and homogeneous LV activation pattern. PMID- 20025706 TI - Is antiarrhythmia device implantation safe under dual antiplatelet therapy? AB - BACKGROUND: Device implantations in patients on dual antiplatelet-therapy (DA therapy) continue to rise. The aim of our study was to compile and analyze data on complications of antiarrhythmia device implantation under DA-therapy. METHODS: We prospectively collected data on all device implantations in our department from January 2008 until February 2009. The control group was comprised of patients on acetylsalicylic acid alone or no antiplatelet medication at all (318 subjects). The DA-therapy group consisted of 109 patients of whom 71 were analyzed retrospectively (implantations from 2002 to 2007). RESULTS: Procedure times were significantly longer in DA-therapy patients receiving a pacemaker for the first time. In contrast, procedure times did not differ significantly between the two study groups for implantable cardioverter defibrillator (ICD) implantations and for pacemaker replacements. Fluid losses via drainage systems and drainage times were significantly increased in the DA-therapy group as compared with the control group after pacemaker but not after ICD implantations. Importantly, there were no significant differences in complication rates, particularly the hematoma rate, between the DA-therapy and the control group. CONCLUSIONS: When drainage systems are used, antiarrhythmia device implantation is safe and can be performed without significantly increased risk of clinically relevant hematoma in patients on continued DA-therapy. (PACE 2010; 394-399). PMID- 20025707 TI - Persistent atrial fibrillation converts to common type atrial flutter during CFAE ablation. AB - BACKGROUND: Catheter ablation of persistent and long-standing persistent atrial fibrillation (AF) is still challenging. So far different ablation techniques have been reported, including pulmonary vein isolation, additional linear lesions, ablation of complex fractionated atrial electrograms (CFAE), and combinations of these techniques. During ablation of CFAE, the occurrence of left atrial (LA) tachycardia is well known. The occurrence of right atrial flutter on the other hand is less well described. METHODS: Here, we report three patients who had been ablated because of symptomatic persistent atrial fibrillation. SUMMARY: In all patients, AF changed into a cavotricuspid isthmus = dependent right atrial flutter during ablation of CFAE in the LA. PMID- 20025709 TI - Qualitative research provides insight into quantitative quality of life measurement. PMID- 20025708 TI - Dual variation in SCN5A and CACNB2b underlies the development of cardiac conduction disease without Brugada syndrome. AB - BACKGROUND: Inherited loss of function mutations in SCN5A have been linked to overlapping syndromes including cardiac conduction disease and Brugada syndrome (BrS). The mechanisms responsible for the development of one without the other are poorly understood. METHODS: Direct sequencing was performed in a family with cardiac conduction disease. Wild-type (WT) and mutant channels were expressed in TSA201 cells for electrophysiological study. Green fluorescent protein (GFP) fused WT or mutant genes were used to assess channel trafficking. RESULTS: A novel SCN5A mutation, P1008S, was identified in all family members displaying first-degree atrioventricular block, but not in unaffected family members nor in 430 reference alleles. Peak P1008S current was 11.77% of WT (P < 0.001). Confocal microscopy showed that WT channels tagged with GFP were localized on the cell surface, whereas GFP-tagged P1008S channels remained trapped in intracellular organelles. Trafficking could be rescued by incubation at room temperature, but not by incubation with mexiletine (300 muM) at 37 degrees C. We also identified a novel polymorphism (D601E) in CACNB2b that slowed inactivation of L-type calcium current (I(Ca,L)), significantly increased total charge. Using the Luo-Rudy action potential (AP) model, we show that the reduction in sodium current (I(Na)) can cause loss of the right ventricular epicardial AP dome in the absence but not in the presence of the slowed inactivation of I(Ca,L). Slowed conduction was present in both cases. CONCLUSIONS: Our results suggest genetic variations leading to a loss-of-function in I(Na) coupled with a gain of function in I(Ca,L) may underlie the development of cardiac conduction disease without BrS. PMID- 20025710 TI - Pathophysiology and clinical implications of cardiac memory. AB - Altering the pattern of activation of the ventricle causes remodeling of the mechanical and electrical properties of the myocardium. The electrical remodeling is evident on the surface electrocardiogram as significant change in T-wave polarity following altered activation; this phenomenon is ascribed to as "T-wave memory" or "cardiac memory." The electrophysiological remodeling following altered activation is characterized by distinct changes in regions proximal (early-activated) versus distal (late-activated) to the site of altered activation. The early-activated region exhibits marked attenuation of epicardial phase 1 notch due to reduced expression of the transient outward potassium current (I(to)). This is attributed to electrotonic changes during altered activation, and angiotensin-mediated regulation of Kv4.3 (the pore-forming alpha subunit responsible for I(to)). The late-activated region exhibits the most significant action potential prolongation due to markedly increased mechanical strain through a mechano-electrical feedback mechanism. Consequently, regionally heterogeneous action potential remodeling occurs following altered activation. This enhances regional repolarization gradients that underlie the electrophysiological basis for T-wave memory. Further, recent clinical studies highlight detrimental consequences of altered activation including worsening mechanical function and increased susceptibility to arrhythmias. Future studies to identify molecular mechanisms that link electrotonic and mechanical strain induced changes to cellular electrophysiology will provide important insights into the role of altered activation in regulating cardiac repolarization and arrhythmogenesis. PMID- 20025711 TI - Melanoma from bench to bedside: meeting report from the 6th international melanoma congress. PMID- 20025712 TI - Melanocytes don't always take the high road. PMID- 20025713 TI - Anatomy of the left atrium for interventional electrophysiologists. AB - Increasingly, interventional procedures require accessing the left atrium from the inside of the heart as well as from the pericardial space. The right phrenic nerve running along the fibrous pericardium is close to the atrial insertion of the right superior pulmonary vein while the left phrenic nerve passes over the left atrial appendage. Posteriorly, the esophagus descends adjacent to the fibrous pericardium covering the posterior and postero-inferior walls of the left atrium. The component parts of the left atrium are reviewed with emphasis on the structure of the atrial septum, the left atrial ridge, the mitral isthmus, and the left atrial walls. Although the atrial walls are mainly smooth, pits and crevices are common in the region of the mitral isthmus and the vicinity of the os of the atrial appendage. The muscular rim around the valve of the oval fossa delimits the extent of the true atrial septum. Interatrial muscular connections exist at the septum, along Bachmann's bundle and also at the muscular sleeves of the coronary sinus and pulmonary veins. Anatomical features relevant to interventional electrophysiologists are highlighted. PMID- 20025714 TI - Linear ablation with duty-cycled radiofrequency energy at the cavotricuspid isthmus. AB - BACKGROUND: Multielectrode catheters using duty-cycled radiofrequency (RF) have been developed to treat atrial fibrillation (AF). Many of these patients also have atrial flutter. Therefore, a linear multielectrode has been developed using the same RF energy. OBJECTIVE: The concept and acute results of linear ablation using duty-cycled RF were tested in the cavotricuspid isthmus (CTI). METHODS: The CTI was targeted in 75 patients, in 68 (90%) among them as an adjunct to AF ablation with the same technology. A linear electrode catheter with a 4-mm tip and five 2-mm ring electrodes was connected to a generator titrating duty-cycled RF at 20-45 W up to a target temperature of 70 degrees C in 1:1 unipolar/bipolar mode. RESULTS: During a mean procedure time of 20 +/- 12 minutes, complete CTI block was achieved by 4 +/- 3 applications of duty-cycled RF in 69 (92%) patients. No more than three RF applications were necessary in 60% of patients. During the initial learning curve, standard RF had to be used in five (7%) patients. Complete block was not achieved in one patient with frequent episodes of AF. Char was observed in five (7%) patients with poor electrode cooling; consequently, the temperature ramp-up was slowed and manually turned off in the event of low-power delivery. Two groin hematomas occurred; otherwise, no clinical complications were observed. CONCLUSION: Multielectrode catheters delivering duty cycled RF can effectively ablate the CTI with few RF applications with promising acute results. Further modifications are necessary to improve catheter steering and prevent char formation. (PACE 2010; 444-450). PMID- 20025715 TI - Heart rate-dependence of QTc intervals assessed by different correction methods in patients with normal or prolonged repolarization. AB - BACKGROUND: There is a continuing debate about the optimal method for QT interval adjustment to heart rate changes. We evaluated the heart rate dependence of QTc intervals derived from five different QT correction methods. METHODS: Study patients (n = 123, age 68 +/- 11 years) were dual-chamber device recipients with baseline normal or prolonged QT interval who had preserved intrinsic ventricular activation with narrow QRS complexes. Patients were classified to either Normal QT (n = 69) or Prolonged-QT (n = 54) groups. Serial QT intervals were recorded at baseline (52 +/- 3 beats per minute) and following atrial pacing stages at 60, 80, and 100 beats per minute. The QTc formulae of Bazett, Fridericia, Sagie Framingham, Hodges, and Karjalainen-Nomogram were applied to assess the effect of heart rate on the derived QTc values by using linear mixed-effects models. RESULTS: Heart rate had a significant effect on QTc regardless of the formula used (P < 0.05 for all formulae). The Bazett's formula demonstrated the highest QTc variability across heart rate stages (highest F values) in both patient groups (in the total cohort, F = 175.9). In the following rank order, the formulae Hodges, Karjalainen-Nomogram, Sagie-Framingham, and Fridericia showed similar QTc heart rate dependence at both slower and faster heart rates in both patient groups (F = 21.8, 25.6, 28.8, 36.9, in the total cohort, respectively). CONCLUSIONS: Of the studied QTc formulae, the Bazett appeared the most heart rate dependent. Our results suggest the use of Hodges and the Karjalainen-Nomogram secondly to ensure least heart rate dependence of QTc intervals in patients with either normal or prolonged repolarization. PMID- 20025716 TI - Frequent recurrent polymorphic ventricular tachycardia during sleep due to managed ventricular pacing. AB - We report a case of a patient with an implantable cardioverter defibrillator and no prior history of heart block with managed ventricular pacing (MVP) programmed who had frequent recurrent episodes of polymorphic ventricular tachycardia. All of the episodes were initiated by transient atrioventricular block which resulted in short-long-short sequences permitted by MVP. This case illustrates that MVP should be used with caution not only in patients with complete heart block, but also in patients at risk for brief heart block due to such states as hypervagatonia due to sleep apnea. PMID- 20025717 TI - Accessory pathway reciprocating tachycardia involving twin AV nodes in a patient with atrioventricular discordance and mitral atresia. AB - The atrioventricular (AV) conduction system in AV discordance remains unclear, especially in cases with complex cardiac anomaly. We report a case of accessory pathway reciprocating tachycardia in atrioventricular discordance (AVD) and mitral atresia with twin AV nodes. In this case, the anterior AV node was located along the atretic mitral valve. The anterior AV node was involved in tachycardia and the posterior AV node acted as a bystander during tachycardia. The anterior AV node in AVD can be located along the atretic mitral valve, and one of twin AV nodes might act as a bystander during AV reciprocating tachycardia. PMID- 20025718 TI - Supraclavicular vein approach for upgrading an implantable cardioverter defibrillator to a biventricular device. AB - Total occlusion of the left subclavian vein was found in a 52-year-old patient, 5 years after implantation of an implantable cardioverter defibrillator (ICD). During replacement, the ICD was upgraded to a biventricular device for worsening of the patient's congestive heart failure to New York Heart Association class III. Insertion of the left ventricular lead in the ipsilateral vein system was successfully achieved by using the supraclavicular approach, enabling puncturing of the left subclavian vein medially to the obstruction. PMID- 20025719 TI - Complete atrioventricular block during ablation of atrial flutter. AB - We report the case of a 51-year-old patient who developed a complete atrioventricular (AV) block during the isthmic radiofrequency catheter ablation of a typical atrial flutter. The cause was an acute occlusion of the segment three of the right coronary artery. His recanalization was associated with the immediate restoration of a normal AV conduction. The complication is exceptional (one of 740 consecutive atrial flutter ablations). (PACE 2010; 516-519). PMID- 20025720 TI - Left atrial voltage during atrial fibrillation in paroxysmal and persistent atrial fibrillation patients. AB - BACKGROUND: Left atrial (LA) endocardial voltage characteristics assessed during atrial fibrillation (AF) have not been previously compared in different AF types. This study was aimed at investigating the LA voltages and volumes in patients with paroxysmal and persistent AF. METHODS: LA electroanatomic voltage maps acquired during AF were compared between consecutive patients without major structural heart disease undergoing first catheter ablation for paroxysmal AF (n = 100) or persistent AF (n = 100). The groups were comparable in baseline clinical characteristics. RESULTS: Patients with persistent AF presented with lower median LA voltage (median 0.41, interquartile range [IQR] 0.31-0.51 mV versus median 0.99, IQR 0.47-1.56 mV; P < 0.001), and maximum LA voltage (4.07 +/ 1.76 vs 6.42 +/- 2.16 mV; P < 0.001). They also had a higher proportion of the LA points exhibiting voltage <0.2 mV (30 +/- 20 vs 12 +/- 11%; P < 0.001) and voltage 0.2-1.0 mV (55 +/- 15 vs 42 +/- 19%; P < 0.001). They further displayed higher LA volume/body surface area (75 +/- 16 vs 58 +/- 13 mL/m(2); P < 0.001). In the multivariate regression model, both LA voltage (P < 10(-9)) and LA volume (P < 10(-5)) were significant determinants of AF type. CONCLUSION: Patients with persistent AF had significantly lower LA voltage compared with patients with paroxysmal AF even after adjustment for differences in indexed LA volume. LA voltage represents an independent covariate of clinical manifestation of AF. PMID- 20025721 TI - Quality assessment of DNA derived from up to 30 years old formalin fixed paraffin embedded (FFPE) tissue for PCR-based methylation analysis using SMART-MSP and MS HRM. AB - BACKGROUND: The High Resolution Melting (HRM) technology has recently been introduced as a rapid and robust analysis tool for the detection of DNA methylation. The methylation status of multiple tumor suppressor genes may serve as biomarkers for early cancer diagnostics, for prediction of prognosis and for prediction of response to treatment. Therefore, it is important that methodologies for detection of DNA methylation continue to evolve. Sensitive Melting Analysis after Real Time - Methylation Specific PCR (SMART-MSP) and Methylation Sensitive - High Resolution Melting (MS-HRM) are two methods for single locus DNA methylation detection based on HRM. METHODS: Here, we have assessed the quality of DNA extracted from up to 30 years old Formalin Fixed Paraffin Embedded (FFPE) tissue for DNA methylation analysis using SMART-MSP and MS-HRM. The quality assessment was performed on DNA extracted from 54 Non-Small Cell Lung Cancer (NSCLC) samples derived from FFPE tissue, collected over 30 years and grouped into five years intervals. For each sample, the methylation levels of the CDKN2A (p16) and RARB promoters were estimated using SMART-MSP and MS-HRM assays designed to assess the methylation status of the same CpG positions. This allowed for a direct comparison of the methylation levels estimated by the two methods for each sample. RESULTS: CDKN2A promoter methylation levels were successfully determined by SMART-MSP and MS-HRM in all 54 samples. Identical methylation estimates were obtained by the two methods in 46 of the samples. The methylation levels of the RARB promoter were successfully determined by SMART-MSP in all samples. When using MS-HRM to assess RARB methylation five samples failed to amplify and 15 samples showed a melting profile characteristic for heterogeneous methylation. Twenty-seven of the remaining 34 samples, for which the methylation level could be estimated, gave the same result as observed when using SMART-MSP. CONCLUSION: MS-HRM and SMART MSP can be successfully used for single locus methylation studies using DNA derived from up to 30 years old FFPE tissue. Furthermore, it can be expected that MS-HRM and SMART-MSP will provide similar methylation estimates when assays are designed to analyze the same CpG positions. PMID- 20025722 TI - RNA degradation compromises the reliability of microRNA expression profiling. AB - BACKGROUND: MicroRNAs are small non-coding RNAs that post-transcriptionally regulate gene expression and their expression is frequently altered in human diseases, including cancer. To correlate clinically relevant parameters with microRNA expression, total RNA is frequently prepared from samples that were archived for various time periods in frozen tissue banks but, unfortunately, RNA integrity is not always preserved in these frozen tissues. Here, we investigate whether experimentally induced RNA degradation affects microRNA expression profiles. RESULTS: Tissue samples were maintained on ice for defined time periods prior to total RNA extraction, which resulted in different degrees of RNA degradation. MicroRNA expression was then analyzed by microarray analysis (miCHIP) or microRNA-specific real-time quantitative PCR (miQPCR). Our results demonstrate that the loss of RNA integrity leads to in unpredictability of microRNA expression profiles for both, array-based and miQPCR assays. CONCLUSION: MicroRNA expression cannot be reliably profiled in degraded total RNA. For the profiling of microRNAs we recommend use of RNA samples with a RNA integrity number equal to or above seven. PMID- 20025723 TI - Identifying significant genetic regulatory networks in the prostate cancer from microarray data based on transcription factor analysis and conditional independency. AB - BACKGROUND: Prostate cancer is a world wide leading cancer and it is characterized by its aggressive metastasis. According to the clinical heterogeneity, prostate cancer displays different stages and grades related to the aggressive metastasis disease. Although numerous studies used microarray analysis and traditional clustering method to identify the individual genes during the disease processes, the important gene regulations remain unclear. We present a computational method for inferring genetic regulatory networks from micorarray data automatically with transcription factor analysis and conditional independence testing to explore the potential significant gene regulatory networks that are correlated with cancer, tumor grade and stage in the prostate cancer. RESULTS: To deal with missing values in microarray data, we used a K nearest-neighbors (KNN) algorithm to determine the precise expression values. We applied web services technology to wrap the bioinformatics toolkits and databases to automatically extract the promoter regions of DNA sequences and predicted the transcription factors that regulate the gene expressions. We adopt the microarray datasets consists of 62 primary tumors, 41 normal prostate tissues from Stanford Microarray Database (SMD) as a target dataset to evaluate our method. The predicted results showed that the possible biomarker genes related to cancer and denoted the androgen functions and processes may be in the development of the prostate cancer and promote the cell death in cell cycle. Our predicted results showed that sub-networks of genes SREBF1, STAT6 and PBX1 are strongly related to a high extent while ETS transcription factors ELK1, JUN and EGR2 are related to a low extent. Gene SLC22A3 may explain clinically the differentiation associated with the high grade cancer compared with low grade cancer. Enhancer of Zeste Homolg 2 (EZH2) regulated by RUNX1 and STAT3 is correlated to the pathological stage. CONCLUSIONS: We provide a computational framework to reconstruct the genetic regulatory network from the microarray data using biological knowledge and constraint-based inferences. Our method is helpful in verifying possible interaction relations in gene regulatory networks and filtering out incorrect relations inferred by imperfect methods. We predicted not only individual gene related to cancer but also discovered significant gene regulation networks. Our method is also validated in several enriched published papers and databases and the significant gene regulatory networks perform critical biological functions and processes including cell adhesion molecules, androgen and estrogen metabolism, smooth muscle contraction, and GO-annotated processes. Those significant gene regulations and the critical concept of tumor progression are useful to understand cancer biology and disease treatment. PMID- 20025724 TI - Patient-centred tuberculosis treatment delivery under programmatic conditions in Tanzania: a cohort study. AB - BACKGROUND: Directly observed therapy (DOT) remains the cornerstone of the global tuberculosis (TB) control strategy. Tanzania, one of the 22 high-burden countries regarding TB, changed the first-line treatment regimen to contain rifampicin containing fixed-dose combination for the full 6 months of treatment. As daily health facility-based DOT for this long period is not feasible for the patient, nor for the health system, Tanzania introduced patient centred treatment (PCT). PCT allows patients to choose for daily DOT at a health facility or at their home by a supporter of choice. The introduction of fixed dose combinations in the intensive and continuation phase made PCT feasible by eliminating the risk of selective drug taking by patients and reducing the number of tablets to be taken. The approach was tested in three districts with the objective to assess the effect of this strategy on TB treatment outcomes METHODS: Cohort analysis comparing patients treated under the PCT strategy (registered April-September 2006) with patients treated under health-facility-based DOT (registered April September 2005). The primary outcome was the cure rate. Differences were assessed by calculating the risk ratios. Associations between characteristics of the supporters and treatment outcomes in the group of patients opting for home-based DOT were assessed through logistic regression. RESULTS: In the PCT cohort there were 1208 patients and 1417 were included in the historic cohort. There was no significant difference in cure rates between the cohorts (risk ratio [RR]: 1.06; 95% confidence interval [CI]: 0.96-1.16). In the PCT cohort, significantly more patients had successful treatment (cure or treatment completed; RR: 1.10; 95%CI: 1.01-1.15). There were no characteristics of supporters that were associated with treatment outcome. CONCLUSION: The PCT approach showed similar cure rates and better treatment success rates compared to daily health-facility DOT. The results indicate that there are no specific prerequisites for the supporter chosen by the patient. The programmatic setting of the study lends strong support for scaling up of TB treatment observation outside the health facility. PMID- 20025725 TI - Digging deeper: quality of patient-provider communication across Hispanic subgroups. AB - BACKGROUND: Recent research suggests that ethnic subgroup designation plays an important role in health-related disparities among Hispanics. Our objective was to examine the influence of Hispanics' self-reported ethnic subgroup designation on perceptions of their health care providers' communication behaviors. METHODS: Cross-sectional analysis of the 2005 Medical Expenditure Panel Survey (MEPS). Participants included non-institutionalized Hispanics (n = 5197; US population estimate = 27,070,906), aged > or = 18 years, reporting visiting a health care provider within the past 12 months. Six (n = 6) items were used to capture respondents' perceptions of their health care providers' communication behaviors. RESULTS: After controlling for socio-demographic covariates, compared to Other Hispanics (reference group), very few differences in perceptions of health care providers communication emerged across ethnic subgroups. Puerto Ricans were more likely to report that their health care provider "always" showed respect for what they had to say (OR = 2.16, 95% CI 1.16-4.03). Both Puerto Ricans (OR = 2.28, 95% CI 1.06-4.92) and Mexicans (OR = 1.88, 95% CI 1.02-3.46) were more likely to indicate that their health care provider "always" spent enough time with them as compared to Other Hispanics. CONCLUSIONS: We observed very few differences among Hispanics respondents in their perceived quality of interactions with health care providers as a function of their ethnic subgroup designation. While our findings somewhat contradict previous research, they do suggest that other underlying factors may influence the quality of perceived interactions with health care providers. PMID- 20025726 TI - Subjective versus objective risk in genetic counseling for hereditary breast and/or ovarian cancers. AB - BACKGROUND: Despite the fact that genetic counseling in oncology provides information regarding objective risks, it can be found a contrast between the subjective and objective risk. The aims of this study were to evaluate the accuracy of the perceived risk compared to the objective risk estimated by the BRCApro computer model and to evaluate any associations between medical, demographic and psychological variables and the accuracy of risk perception. METHODS: 130 subjects were given medical-demographic file, Cancer and Genetic Risk Perception, Hospital Anxiety-Depression Scale. It was also computed an objective evaluation of the risk by the BRCApro model. RESULTS: The subjective risk was significantly higher than objective risk. The risk of tumour was overestimated by 56%, and the genetic risk by 67%. The subjects with less cancer affected relatives significantly overestimated their risk of being mutation carriers and made a more inaccurate estimation than high risk subjects. CONCLUSION: The description of this sample shows: general overestimation of the risk, inaccurate perception compared to BRCApro calculation and a more accurate estimation in those subjects with more cancer affected relatives (high risk subjects). No correlation was found between the levels of perception of risk and anxiety and depression. Based on our findings, it is worth pursuing improved communication strategies about the actual cancer and genetic risk, especially for subjects at "intermediate and slightly increased risk" of developing an hereditary breast and/or ovarian cancer or of being mutation carrier. PMID- 20025727 TI - Efficacy of smoking prevention program 'Smoke-free Kids': study protocol of a randomized controlled trial. AB - BACKGROUND: A strong increase in smoking is noted especially among adolescents. In the Netherlands, about 5% of all 10-year olds, 25% of all 13-year olds and 62% of all 17-year olds report ever smoking. In the U.S., an intervention program called 'Smoke-free Kids' was developed to prevent children from smoking. The present study aims to assess the effects of this home-based smoking prevention program in the Netherlands. METHODS/DESIGN: A randomized controlled trial is conducted among 9 to 11-year old children of primary schools. Participants are randomly assigned to the intervention and control conditions. The intervention program consists of five printed activity modules designed to improve parenting skills specific to smoking prevention and parent-child communication regarding smoking. These modules will include additional sheets with communication tips. The modules for the control condition will include solely information on smoking and tobacco use.Initiation of cigarette smoking (first instance of puffing on a lighted cigarette), susceptibility to cigarette smoking, smoking-related cognitions, and anti-smoking socialization will be the outcome measures. To collect the data, telephone interviews with mothers as well as with their child will be conducted at baseline. Only the children will be examined at post intervention follow-ups (6, 12, 24, and 36 months after the baseline). DISCUSSION: This study protocol describes the design of a randomized controlled trial that will evaluate the effectiveness of a home-based smoking prevention program. We expect that a significantly lower number of children will start smoking in the intervention condition compared to control condition as a direct result of this intervention. If the program is effective, it is applicable in daily live, which will facilitate implementation of the prevention protocol. TRIAL REGISTRATION: Netherlands Trial Register NTR1465. PMID- 20025728 TI - Late treatment with imatinib mesylate ameliorates radiation-induced lung fibrosis in a mouse model. AB - BACKGROUND: We have previously shown that small molecule PDGF receptor tyrosine kinase inhibitors (RTKI) can drastically attenuate radiation-induced pulmonary fibrosis if the drug administration starts at the time of radiation during acute inflammation with present but limited effects against acute inflammation. To rule out interactions of the drug with acute inflammation, we investigated here in an interventive trial if a later drug administration start at a time when the acute inflammation has subsided--has also beneficial antifibrotic effects. METHODS: Whole thoraces of C57BL/6 mice were irradiated with 20 Gy and treated with the RTKI imatinib starting either 3 days after radiation (during acute inflammation) or two weeks after radiation (after the acute inflammation has subsided as demonstrated by leucocyte count). Lungs were monitored and analyzed by clinical, histological and in vivo non-invasive computed tomography as a quantitative measure for lung density and lung fibrosis. RESULTS: Irradiation induced severe lung fibrosis resulting in markedly reduced mouse survival vs. non-irradiated controls. Both early start of imatinib treatment during inflammation and late imatinib start markedly attenuated the development of pulmonary fibrosis as demonstrated by clinical, histological and qualitative and quantitative computed tomography results such as reduced lung density. Both administration schedules resulted in prolonged lifespans. The earlier drug treatment start resulted in slightly stronger beneficial antifibrotic effects along all measured endpoints than the later start. CONCLUSIONS: Our findings show that imatinib, even when administered after the acute inflammation has subsided, attenuates radiation induced lung fibrosis in mice. Our data also indicate that the fibrotic fate is not only determined by the early inflammatory events but rather a complex process in which secondary events at later time points are important. Because of the clinical availability of imatinib or similar compounds, a meaningful attenuation of radiation-induced lung fibrosis in patients seems possible. PMID- 20025729 TI - Inverse tuning of metal binding affinity and protein stability by altering charged coordination residues in designed calcium binding proteins. AB - Ca(2+ )binding proteins are essential for regulating the role of Ca(2+ )in cell signaling and maintaining Ca(2+ )homeostasis. Negatively charged residues such as Asp and Glu are often found in Ca(2+ )binding proteins and are known to influence Ca(2+ )binding affinity and protein stability. In this paper, we report a systematic investigation of the role of local charge number and type of coordination residues in Ca(2+ )binding and protein stability using de novo designed Ca(2+ )binding proteins. The approach of de novo design was chosen to avoid the complications of cooperative binding and Ca(2+)-induced conformational change associated with natural proteins. We show that when the number of negatively charged coordination residues increased from 2 to 5 in a relatively restricted Ca(2+)-binding site, Ca(2+ )binding affinities increased by more than 3 orders of magnitude and metal selectivity for trivalent Ln(3+ )over divalent Ca(2+ )increased by more than 100-fold. Additionally, the thermal transition temperatures of the apo forms of the designed proteins decreased due to charge repulsion at the Ca(2+ )binding pocket. The thermal stability of the proteins was regained upon Ca(2+ )and Ln(3+ )binding to the designed Ca(2+ )binding pocket. We therefore observe a striking tradeoff between Ca(2+)/Ln(3+ )affinity and protein stability when the net charge of the coordination residues is varied. Our study has strong implications for understanding and predicting Ca(2+)-conferred thermal stabilization of natural Ca(2+ )binding proteins as well as for designing novel metalloproteins with tunable Ca(2+ )and Ln(3+ )binding affinity and selectivity.PACS codes: 05.10.-a. PMID- 20025730 TI - Optimization of the dilute maleic acid pretreatment of wheat straw. AB - BACKGROUND: In this study, the dilute maleic acid pretreatment of wheat straw is optimized, using pretreatment time, temperature and maleic acid concentration as design variables. A central composite design was applied to the experimental set up. The response factors used in this study are: (1) glucose benefits from improved enzymatic digestibility of wheat straw solids; (2) xylose benefits from the solubilization of xylan to the liquid phase during the pretreatment; (3) maleic acid replenishment costs; (4) neutralization costs of pretreated material; (5) costs due to furfural production; and (6) heating costs of the input materials. For each response factor, experimental data were fitted mathematically. After data translation to euro/Mg dry straw, determining the relative contribution of each response factor, an economic optimization was calculated within the limits of the design variables. RESULTS: When costs are disregarded, an almost complete glucan conversion to glucose can be reached (90% from solids, 7%-10% in liquid), after enzymatic hydrolysis. During the pretreatment, up to 90% of all xylan is converted to monomeric xylose. Taking cost factors into account, the optimal process conditions are: 50 min at 170 degrees C, with 46 mM maleic acid, resulting in a yield of 65 euro/Mg (megagram = metric ton) dry straw, consisting of 68 euro/Mg glucose benefits (from solids: 85% of all glucan), 17 euro/Mg xylose benefits (from liquid: 80% of all xylan), 17 euro/Mg maleic acid costs, 2.0 euro/Mg heating costs and 0.68 euro/Mg NaOH costs. In all but the most severe of the studied conditions, furfural formation was so limited that associated costs are considered negligible. CONCLUSIONS: After the dilute maleic acid pretreatment and subsequent enzymatic hydrolysis, almost complete conversion of wheat straw glucan and xylan is possible. Taking maleic acid replenishment, heating, neutralization and furfural formation into account, the optimum in the dilute maleic acid pretreatment of wheat straw in this study is 65 euro/Mg dry feedstock. This is reached when process conditions are: 50 min at 170 degrees C, with a maleic acid concentration of 46 mM. Maleic acid replenishment is the most important of the studied cost factors. PMID- 20025731 TI - Decreased mitochondrial DNA content in blood samples of patients with stage I breast cancer. AB - BACKGROUND: Alterations of mitochondrial DNA (mtDNA) have been implicated in carcinogenesis. We developed an accurate multiplex quantitative real-time PCR for synchronized determination of mtDNA and nuclear DNA (nDNA). We sought to investigate whether mtDNA content in the peripheral blood of breast cancer patients is associated with clinical and pathological parameters. METHODS: Peripheral blood samples were collected from 60 patients with breast cancer and 51 age-matched healthy individuals as control. DNA was extracted from peripheral blood for the quantification of mtDNA and nDNA, using a one-step multiplex real time PCR. A FAM labeled MGB probe and primers were used to amplify the mtDNA sequence of the ATP 8 gene, and a VIC labeled MGB probe and primers were employed to amplify the glyceraldehyde-3-phosphate-dehydrogenase gene. mtDNA content was correlated with tumor stage, menstruation status, and age of patients as well as lymph node status and the expression of estrogen receptor (ER), progesterone receptor (PR) and Her-2/neu protein. RESULTS: The content of mtDNA in stage I breast cancer patients was significantly lower than in other stages (overall P = 0.023). Reduced mtDNA was found often in post menopausal cancer group (P = 0.024). No difference in mtDNA content, in regards to age (p = 0.564), lymph node involvement (p = 0.673), ER (p = 0.877), PR (p = 0.763), and Her-2/neu expression (p = 0.335), was observed. CONCLUSION: Early detection of breast cancer has proved difficult and current detection methods are inadequate. In the present study, decreased mtDNA content in the peripheral blood of patients with breast cancer was strongly associated with stage I. The use of mtDNA may have diagnostic value and further studies are required to validate it as a potential biomarker for early detection of breast cancer. PMID- 20025732 TI - Myocardial tissue tagging with cardiovascular magnetic resonance. AB - Cardiovascular magnetic resonance (CMR) is currently the gold standard for assessing both global and regional myocardial function. New tools for quantifying regional function have been recently developed to characterize early myocardial dysfunction in order to improve the identification and management of individuals at risk for heart failure. Of particular interest is CMR myocardial tagging, a non-invasive technique for assessing regional function that provides a detailed and comprehensive examination of intra-myocardial motion and deformation. Given the current advances in gradient technology, image reconstruction techniques, and data analysis algorithms, CMR myocardial tagging has become the reference modality for evaluating multidimensional strain evolution in the human heart. This review presents an in depth discussion on the current clinical applications of CMR myocardial tagging and the increasingly important role of this technique for assessing subclinical myocardial dysfunction in the setting of a wide variety of myocardial disease processes. PMID- 20025734 TI - Array-based DNA methylation profiling of primary lymphomas of the central nervous system. AB - BACKGROUND: Although primary lymphomas of the central nervous system (PCNSL) and extracerebral diffuse large B-cell lymphoma (DLBCL) cannot be distinguished histologically, it is still a matter of debate whether PCNSL differ from systemic DLBCL with respect to their molecular features and pathogenesis. Analysis of the DNA methylation pattern might provide further data distinguishing these entities at a molecular level. METHODS: Using an array-based technology we have assessed the DNA methylation status of 1,505 individual CpG loci in five PCNSL and compared the results to DNA methylation profiles of 49 DLBCL and ten hematopoietic controls. RESULTS: We identified 194 genes differentially methylated between PCNSL and normal controls. Interestingly, Polycomb target genes and genes with promoters showing a high CpG content were significantly enriched in the group of genes hypermethylated in PCNSL. However, PCNSL and systemic DLBCL did not differ in their methylation pattern. CONCLUSIONS: Based on the data presented here, PCNSL and DLBCL do not differ in their DNA methylation pattern. Thus, DNA methylation analysis does not support a separation of PCNSL and DLBCL into individual entities. However, PCNSL and DLBCL differ in their DNA methylation pattern from non- malignant controls. PMID- 20025733 TI - Functional genomics of pH homeostasis in Corynebacterium glutamicum revealed novel links between pH response, oxidative stress, iron homeostasis and methionine synthesis. AB - BACKGROUND: The maintenance of internal pH in bacterial cells is challenged by natural stress conditions, during host infection or in biotechnological production processes. Comprehensive transcriptomic and proteomic analyses has been conducted in several bacterial model systems, yet questions remain as to the mechanisms of pH homeostasis. RESULTS: Here we present the comprehensive analysis of pH homeostasis in C. glutamicum, a bacterium of industrial importance. At pH values between 6 and 9 effective maintenance of the internal pH at 7.5 +/- 0.5 pH units was found. By DNA microarray analyses differential mRNA patterns were identified. The expression profiles were validated and extended by 1D-LC-ESI MS/MS based quantification of soluble and membrane proteins. Regulators involved were identified and thereby participation of numerous signaling modules in pH response was found. The functional analysis revealed for the first time the occurrence of oxidative stress in C. glutamicum cells at neutral and low pH conditions accompanied by activation of the iron starvation response. Intracellular metabolite pool analysis unraveled inhibition of the TCA and other pathways at low pH. Methionine and cysteine synthesis were found to be activated via the McbR regulator, cysteine accumulation was observed and addition of cysteine was shown to be toxic under acidic conditions. CONCLUSIONS: Novel limitations for C. glutamicum at non-optimal pH values were identified by a comprehensive analysis on the level of the transcriptome, proteome, and metabolome indicating a functional link between pH acclimatization, oxidative stress, iron homeostasis, and metabolic alterations. The results offer new insights into bacterial stress physiology and new starting points for bacterial strain design or pathogen defense. PMID- 20025735 TI - Phylogenomics of the oxidative phosphorylation in fungi reveals extensive gene duplication followed by functional divergence. AB - BACKGROUND: Oxidative phosphorylation is central to the energy metabolism of the cell. Due to adaptation to different life-styles and environments, fungal species have shaped their respiratory pathways in the course of evolution. To identify the main mechanisms behind the evolution of respiratory pathways, we conducted a phylogenomics survey of oxidative phosphorylation components in the genomes of sixty fungal species. RESULTS: Besides clarifying orthology and paralogy relationships among respiratory proteins, our results reveal three parallel losses of the entire complex I, two of which are coupled to duplications in alternative dehydrogenases. Duplications in respiratory proteins have been common, affecting 76% of the protein families surveyed. We detect several instances of paralogs of genes coding for subunits of respiratory complexes that have been recruited to other multi-protein complexes inside and outside the mitochondrion, emphasizing the role of evolutionary tinkering. CONCLUSIONS: Processes of gene loss and gene duplication followed by functional divergence have been rampant in the evolution of fungal respiration. Overall, the core proteins of the respiratory pathways are conserved in most lineages, with major changes affecting the lineages of microsporidia, Schizosaccharomyces and Saccharomyces/Kluyveromyces due to adaptation to anaerobic life-styles. We did not observe specific adaptations of the respiratory metabolism common to all pathogenic species. PMID- 20025736 TI - A structured registration program can be validly used for quality assessment in general practice. AB - BACKGROUND: Patient information, medical history, clinical outcomes and demographic information, can be registered in different ways in registration programs. For evaluation of diabetes care, data can easily be extracted from a structured registration program (SRP). The usability of data from this source depends on the agreement of this data with that of the usual data registration in the electronic medical record (EMR).Aim of the study was to determine the comparability of data from an EMR and from an SRP, to determine whether the use of SRP data for quality assessment is justified in general practice. METHODS: We obtained 196 records of diabetes mellitus patients in a sample of general practices in the Netherlands. We compared the agreement between the two programs in terms of laboratory and non-laboratory parameters. Agreement was determined by defining accordance between the programs in absent and present registrations, accordance between values of registrations, and whether the differences found in values were also a clinically relevant difference. RESULTS: No differences were found in the occurrence of registration (absent/present) in the SRP and EMR for all the laboratory parameters. Smoking behaviour, weight and eye examination were registered significantly more often in the SRP than in the EMR. In the EMR, blood pressure was registered significantly more often than in the SRP. Data registered in the EMR and in the SRP had a similar clinical meaning for all parameters (laboratory and non-laboratory). CONCLUSIONS: Laboratory parameters showed good agreement and non-laboratory acceptable agreement of the SRP with the EMR. Data from a structured registration program can be used validly for research purposes and quality assessment in general practice. PMID- 20025737 TI - The effect of proteoglycans inhibited by RNA interference on metastatic characters of human salivary adenoid cystic carcinoma. AB - BACKGROUND: Salivary adenoid cystic carcinoma (SACC) is one of the most common malignancies of salivary gland. Recurrence or/and early metastasis is its biological properties. In SACC, neoplastic myoepithelial cells secrete proteoglycans unconventionally full of the cribriform or tubular and glandular structures of SACC. Literatures have demonstrated that extracellular matrix provided an essential microenvironment for the biological behavior of SACC. However, there is rare study of the effect of proteoglycans on the potential metastasis of SACC.In this study, human xylosyltransferase-I (XTLY-I) gene, which catalyzes the rate-limited step of proteoglycans biosynthesis, was knocked down by RNA interference (RNAi) to inhibit the proteoglycans biosynthesis in SACC cell line with high tendency of lung metastasis (SACC-M). The impact of down-regulated proteoglycans on the metastasis characters of SACC-M cells was analyzed and discussed. This research could provide a new idea for the clinical treatment of SACC. METHODS: The eukaryotic expression vector of short hairpin RNA (shRNA) targeting XTLY-I gene was constructed and transfected into SACC-M cells. A stably transfectant cell line named SACC-M-WJ4 was isolated. The XTLY-I expression was measured by real-time PCR and Western blot; the reduction of proteoglycans was measured. The invasion and metastasis of SACC-M-WJ4 cells were detected; the effect of down-regulated proteoglycans on the potential lung metastasis of nude mice was observed, respectively. RESULTS: The shRNA plasmid targeting XTLY-I gene showed powerful efficiency of RNAi. The mRNA level of target gene decreased by 86.81%, the protein level was decreased by 80.10%, respectively. The silence of XTLY-I gene resulted in the reduction of proteoglycans significantly in SACC-M WJ4 cells. The inhibitory rate of proteoglycans was 58.17% (24 h), 66.06% (48 h), 57.91% (72 h), 59.36% (96 h), and 55.65% (120 h), respectively. The reduction of proteoglycans suppressed the adhesion, invasion and metastasis properties of SACC M cells, and decreased the lung metastasis of SACC-M cells markedly either. CONCLUSION: The data suggested that the silence of XTLY-I gene in SACC-M cells could suppress proteoglycans biosynthesis and secretion significantly. The reduction of proteoglycans inhibited cell adhesion, invasion and metastasis of SACC-M cells. There is a close relationship between proteoglycans and the biological behavior of SACC. PMID- 20025738 TI - Anaplasma phagocytophilum in Danish sheep: confirmation by DNA sequencing. AB - BACKGROUND: The presence of Anaplasma phagocytophilum, an Ixodes ricinus transmitted bacterium, was investigated in two flocks of Danish grazing lambs. Direct PCR detection was performed on DNA extracted from blood and serum with subsequent confirmation by DNA sequencing. METHODS: 31 samples obtained from clinically normal lambs in 2000 from Fussingo, Jutland and 12 samples from ten lambs and two ewes from a clinical outbreak at Feddet, Zealand in 2006 were included in the study. Some of the animals from Feddet had shown clinical signs of polyarthritis and general unthriftiness prior to sampling. DNA extraction was optimized from blood and serum and detection achieved by a 16S rRNA targeted PCR with verification of the product by DNA sequencing. RESULTS: Five DNA extracts were found positive by PCR, including two samples from 2000 and three from 2006. For both series of samples the product was verified as A. phagocytophilum by DNA sequencing. CONCLUSIONS: A. phagocytophilum was detected by molecular methods for the first time in Danish grazing lambs during the two seasons investigated (2000 and 2006). PMID- 20025739 TI - Fluvoxamine monotherapy for psychotic depression: the potential role of sigma-1 receptors. AB - BACKGROUND: Psychotic depression is a clinical subtype of major depressive disorder. A number of clinical studies have demonstrated the efficacy of the combination of an antidepressant (for example, a tricyclic antidepressant or selective serotonin reuptake inhibitor (SSRI)) and an atypical antipsychotic or electroconvulsive therapy in treating psychotic depression. In some cases, the clinician or patient may prefer to avoid antipsychotic drugs altogether because of the risk of extrapyramidal side effects (EPS) in patients with psychotic depression treated with these drugs. METHODS: We report five cases where fluvoxamine monotherapy was effective in the patients with psychotic depression. RESULTS: The scores on the Hamilton Depression (HAM-D) scale and the Brief Psychiatric Rating Scale (BPRS) in the five patients with psychotic depression were reduced after fluvoxamine monotherapy. CONCLUSION: Doctors should consider fluvoxamine monotherapy as an alternative approach in treating psychotic depression because it avoids the risk of EPS from antipsychotic drugs. PMID- 20025740 TI - Mutation analysis of the AATF gene in breast cancer families. AB - BACKGROUND: About 5-10% of breast cancer is due to inherited disease predisposition. Many previously identified susceptibility factors are involved in the maintenance of genomic integrity. AATF plays an important role in the regulation of gene transcription and cell proliferation. It induces apoptosis by associating with p53. The checkpoint kinases ATM/ATR and CHEK2 interact with and phosphorylate AATF, enhancing its accumulation and stability. Based on its biological function, and direct interaction with several known breast cancer risk factors, AATF is a good candidate gene for being involved in heritable cancer susceptibility. METHODS: Here we have screened the entire coding region of AATF in affected index cases from 121 Finnish cancer families for germline defects, using conformation sensitive gel electrophoresis and direct sequencing. RESULTS: Altogether seven different sequence changes were observed, one missense variant and six intronic ones. Based on the in silico analyses of these sequence alterations, as well as their occurrence in cases and controls, none of them, however, were predicted to be pathogenic. CONCLUSIONS: To our knowledge, this is the first study reporting the mutation screening of the AATF gene in familial breast cancer cases. No evidence for the association with breast cancer was observed. PMID- 20025741 TI - Prophylactic and therapeutic activity of fully human monoclonal antibodies directed against influenza A M2 protein. AB - Influenza virus infection is a prevalent disease in humans. Antibodies against hemagglutinin have been shown to prevent infection and hence hemagglutinin is the major constituent of current vaccines. Antibodies directed against the highly conserved extracellular domain of M2 have also been shown to mediate protection against Influenza A infection in various animal models. Active vaccination is generally considered the best approach to combat viral diseases. However, passive immunization is an attractive alternative, particularly in acutely exposed or immune compromized individuals, young children and the elderly. We recently described a novel method for the rapid isolation of natural human antibodies by mammalian cell display. Here we used this approach to isolate human monoclonal antibodies directed against the highly conserved extracellular domain of the Influenza A M2 protein. The identified antibodies bound M2 peptide with high affinities, recognized native cell-surface expressed M2 and protected mice from a lethal influenza virus challenge. Moreover, therapeutic treatment up to 2 days after infection was effective, suggesting that M2-specific monoclonals have a great potential as immunotherapeutic agents against Influenza infection. PMID- 20025742 TI - Epidemiological investigations of human rabies in China. AB - BACKGROUND: The epidemic of rabies showed a rising trend in China in recent years. To identify the potential factors involved in the emergence, we investigated and analyzed the status and characteristics of human rabies between 1996 and 2008. Moreover, the status of rabies infection and vaccination in dogs, and prophylaxis of humans after rabies exposure were analyzed. METHODS: Human rabies data in China between 1996 and 2008 collected from the annual reports of Chinese Center for Disease Control and Prevention (China CDC) were analyzed. To investigate the status of dogs and postexposure prophylaxis (PEP) of humans, brain specimens of domestic dogs were collected and detected, and the demographic details, exposure status and PEP of rabies patients were obtained in 2005 and 2006 in Guangxi, Hunan and Guizhou provinces. RESULTS: The results showed 19,806 human rabies cases were reported in China from 1996 to 2008, with an average of 1,524 cases each year, and the incidence almost was rising rapidly, with the peak in 2007 (3,300 cases). It was notable that nearly 50% of the total rabies cases nationwide were reported in Guangxi, Hunan and Guizhou provinces. In these three provinces, the rabies infection rate in dogs was 2.3%, and 60% investigated cities had a dog vaccination rate of below 70%; among the 315 recorded human cases, 66.3% did not receive any PEP at all, 27.6% received inadequate PEP, and only 6.0% received a full regime of PEP. CONCLUSIONS: In recent years, rabies is reemerging and becoming a major public-health problem in China. Our analysis showed that unsuccessful control of dog rabies and inadequate PEP of patients were the main factors leading to the high incidence of human rabies in China, then there are following suggestions: (1) Strict control of free-ranging dogs and mandatory rabies vaccination should be enforced. (2)Establishing national animal rabies surveillance network is imperative. (3) PEP should be decided to initiate or withhold according to postmortem diagnosis of the biting animal. (4) The cost of PEP should be decreased or free, especially in rural areas. (5)Education of the public and health care staff should be enhanced. PMID- 20025743 TI - Reasons for non-participation in a parental program concerning underage drinking: a mixed-method study. AB - BACKGROUND: Alcohol consumption among adolescents is a serious public health concern. Research has shown that prevention programs targeting parents can help prevent underage drinking. The problem is that parental participation in these kinds of interventions is generally low. Therefore, the aim of the present study is to examine non-participation in a parental support program aiming to prevent underage alcohol drinking. The Health Belief Model has been used as a tool for the analysis. METHODS: To understand non-participation in a parental program a quasi-experimental mixed-method design was used. The participants in the study were invited to participate in a parental program targeting parents with children in school years 7-9. A questionnaire was sent home to the parents before the program started. Two follow-up surveys were also carried out. The inclusion criteria for the study were that the parents had answered the questionnaire in school year 7 and either of the questionnaires in the two subsequent school years (n = 455). Multinomial logistic regression analysis was used to examine reasons for non-participation. The final follow-up questionnaire included an opened-ended question about reasons for non-participation. A qualitative content analysis was carried out and the two largest categories were included in the third model of the multinomial logistic regression analysis. RESULTS: Educational level was the most important socio-demographic factor for predicting non-participation. Parents with a lower level of education were less likely to participate than those who were more educated. Factors associated with adolescents and alcohol did not seem to be of significant importance. Instead, program-related factors predicted non participation, e.g. parents who did not perceive any need for the intervention and who did not attend the information meeting were more likely to be non participants. Practical issues, like time demands, also seemed to be important. CONCLUSION: To design a parental program that attracts parents independently of educational level seems to be an important challenge for the future as well as program marketing. This is something that must be considered when implementing prevention programs. PMID- 20025744 TI - The influence of insurance status on waiting times in German acute care hospitals: an empirical analysis of new data. AB - BACKGROUND: There is an ongoing debate in Germany about the assumption that patients with private health insurance (PHI) benefit from better access to medical care, including shorter waiting times (Lungen et al. 2008), compared to patients with statutory health insurance (SHI). PROBLEM: Existing analyses of the determinants for waiting times in Germany are a) based on patient self-reports and b) do not cover the inpatient sector. This paper aims to fill both gaps by (i) generating new primary data and (ii) analyzing waiting times in German hospitals. METHODS: We requested individual appointments from 485 hospitals within an experimental study design, allowing us to analyze the impact of PHI versus SHI on waiting times (Asplin et al. 2005). RESULTS: In German acute care hospitals patients with PHI have significantly shorter waiting times than patients with SHI. CONCLUSION: Discrimination in waiting times by insurance status does occur in the German acute hospital sector. Since there is very little transparency in treatment quality in Germany, we do not know whether discrimination in waiting times leads to discrimination in the quality of treatment. This is an important issue for future research. PMID- 20025745 TI - Identification of a novel betaherpesvirus in Mus musculus. AB - Rodent betaherpesviruses vary considerably in genomic content, and these variations can result in a distinct pathogenicity. Therefore, the identification of unknown betaherpesviruses in house mice (Mus musculus), the most important rodent host species in basic research, is of importance. During a search for novel herpesviruses in house mice using herpesvirus consensus PCR and attempts to isolate viruses in tissue culture, we identified a previously unknown betaherpesvirus. The primary PCR search in mouse organs revealed the presence of known strains of murine cytomegalovirus (Murid herpesvirus 1) and of Mus musculus rhadinovirus 1 only. However, the novel virus was detected after incubation of organ pieces in fibroblast tissue culture and subsequent PCR analysis of the supernatants. Long-distance PCR amplification including the DNA polymerase and glycoprotein B genes revealed a 3.4 kb sequence that was similar to sequences of rodent cytomegaloviruses. Pairwise sequence comparisons and phylogenetic analyses showed that this newly identified murine virus is most similar to the English isolate of rat cytomegalovirus, thereby raising the possibility that two distinct CMV lineages have evolved in both Mus musculus and Rattus norvegicus. PMID- 20025746 TI - Isolation and characterization of a novel plasma membrane protein, osteoblast induction factor (obif), associated with osteoblast differentiation. AB - BACKGROUND: While several cell types are known to contribute to bone formation, the major player is a common bone matrix-secreting cell type, the osteoblast. Chondrocytes, which plays critical roles at several stages of endochondral ossification, and osteoblasts are derived from common precursors, and both intrinsic cues and signals from extrinsic cues play critical roles in the lineage decision of these cell types. Several studies have shown that cell fate commitment within the osteoblast lineage requires sequential, stage-specific signaling to promote osteoblastic differentiation programs. In osteoblastic differentiation, the functional mechanisms of transcriptional regulators have been well elucidated, however the exact roles of extrinsic molecules in osteoblastic differentiation are less clear. RESULTS: We identify a novel gene, obif (osteoblast induction factor), encoding a transmembrane protein that is predominantly expressed in osteoblasts. During mouse development, obif is initially observed in the limb bud in a complementary pattern to Sox9 expression. Later in development, obif is highly expressed in osteoblasts at the stage of endochondral ossification. In cell line models, obif is up-regulated during osteoblastic differentiation. Exogenous obif expression stimulates osteoblastic differentiation and obif knockdown inhibits osteoblastic differentiation in preosteblastic MC3T3-E1 cells. In addition, the extracellular domain of obif protein exhibits functions similar to the full-length obif protein in induction of MC3T3-E1 differentiation. CONCLUSIONS: Our results suggest that obif plays a role in osteoblastic differentiation by acting as a ligand. PMID- 20025747 TI - Pre-radiotherapy plasma carotenoids and markers of oxidative stress are associated with survival in head and neck squamous cell carcinoma patients: a prospective study. AB - BACKGROUND: The purpose of this study was to compare plasma levels of antioxidants and oxidative stress biomarkers in head and neck squamous cell carcinoma (HNSCC) patients with healthy controls. Furthermore, the effect of radiotherapy on these biomarkers and their association with survival in HNSCC patients were investigated. METHODS: Seventy-eight HNSCC patients and 100 healthy controls were included in this study. Follow-up samples at the end of radiotherapy were obtained in 60 patients. Fifteen antioxidant biomarkers (6 carotenoids, 4 tocopherols, ascorbic acid, total antioxidant capacity, glutathione redox potential, total glutathione and total cysteine) and four oxidative stress biomarkers (total hydroperoxides, gamma-glutamyl transpeptidase, 8-isoprostagladin F2alpha and ratio of oxidized/total ascorbic acid) were measured in plasma samples. Analysis of Covariance was used to compare biomarkers between patients and healthy controls. Kaplan-Meier plots and Cox' proportional hazards models were used to study survival among patients. RESULTS: Dietary antioxidants (carotenoids, tocopherols and ascorbic acid), ferric reducing antioxidant power (FRAP) and modified FRAP were lower in HNSCC patients compared to controls and dietary antioxidants decreased during radiotherapy. Total hydroperoxides (d-ROMs), a marker for oxidative stress, were higher in HNSCC patients compared to controls and increased during radiotherapy. Among the biomarkers analyzed, high levels of plasma carotenoids before radiotherapy are associated with a prolonged progression-free survival (hazard rate ratio: 0.42, 95% CI: 0.20-0.91, p = 0.03). Additionally, high relative increase in plasma levels of d-ROMs (hazard rate ratio: 0.31, 95% CI: 0.13-0.76, p = 0.01) and high relative decrease in FRAP (hazard rate ratio: 0.42, 95% CI: 0.17-0.998, p = 0.05) during radiotherapy are also positively associated with survival. CONCLUSIONS: Biomarkers of antioxidants and oxidative stress are unfavourable in HNSCC patients compared to healthy controls, and radiotherapy affects many of these biomarkers. Increasing levels of antioxidant biomarkers before radiotherapy and increasing oxidative stress during radiotherapy may improve survival indicating that different factors/mechanisms may be important for survival before and during radiotherapy in HNSCC patients. Thus, the therapeutic potential of optimizing antioxidant status and oxidative stress should be explored further in these patients. PMID- 20025748 TI - Significance of Twist expression and its association with E-cadherin in esophageal squamous cell carcinoma. AB - BACKGROUND: Twist is a basic helix-loop-helix (bHLH) transcriptional factor that has been identified to play an important role in epithelial-mesenchymal transition (EMT)-mediated metastasis through the regulation of E-cadherin expression. However, few authors have examined the expression of Twist and E cadherin and their prognostic value in patients with esophageal squamous cell carcinoma (ESCC). The purpose of this study is to evaluate the clinical significance of Twist and E-cadherin expression in ESCC. METHODS: We immunohistochemically investigated the relationship between their expression and clinicopathological factors including prognosis in surgical specimens of primary tumors in 166 patients with ESCC. RESULTS: The expression rate of high Twist was 42.0% and that of preserved E-cadherin was 40.4%. The expression of high Twist and reduced E-cadherin was significantly associated with depth of tumor invasion, lymph node metastasis, distant nodal metastasis, stage and lymphatic invasion, and poor prognosis. High Twist expression significantly correlated with reduced E cadherin expression. In the preserved E-cadherin group, the 5-year survival rate was better for patients who were low for Twist expression than for those who were high for Twist expression. Multivariate analysis indicated that the combination of low Twist and preserved E-cadherin expression was an independent prognostic factor along with tumor depth, distant nodal metastasis and E-cadherin expression. CONCLUSIONS: Evaluation of Twist and E-cadherin expressions should be useful for determining tumor properties, including prognosis, in patients with ESCC. PMID- 20025749 TI - Relationship between home care service use and changes in the care needs level of Japanese elderly. AB - BACKGROUND: With the introduction of long-term care insurance (LTCI) in Japan, more home care services are available for the community-dwelling elderly. To deliver effective home care services, it is important to know the effects of service use. In this study, as the first step to determine this, we sought to describe different home service use in the sustained/improved group and deteriorated group in their care needs levels, and to report the relationship between the use of home care services and changes in care needs levels. METHODS: The participants included 624 of a total of 1,474 users of LTCI services in one city in Japan. Home care service users were stratified into a 'lower care needs level subgroup' and a 'higher care needs level subgroup' based on the baseline care needs level. Simple statistical comparison and multiple logistic regression analyses in which the change in care needs level was set as a dependent variable were performed. Gender, age, and baseline care needs level were designated as control variables. Home based services were treated as independent variables. In this study, home care services consisted of home help, home bathing services, a visiting nurse, home rehabilitation, nursing home daycare, health daycare, loan of medical devices, respite stay in a nursing home, respite stay in a health care facility, respite stay in a sanatorium-type medical care facility, and medical management by a physician. RESULTS: In the lower care needs level subgroup, age (OR = 1.04, CI, 1.01-1.08), use of respite stay in a nursing home (OR = 2.55; CI, 1.43-4.56), and the number of types of long-term care services (OR = 1.33; CI, 1.02-1.74) used during an 11 month period were significantly related to a deterioration of the user's care needs level. In the higher care needs level subgroup, use of medical management by a physician (OR = 6.99; CI, 1.42-41.25) was significantly related to a deterioration of the user's care needs level. There were no home based services significantly related to sustaining or improving the user's care needs level. CONCLUSION: There were different home service use in two groups (the sustained/improved group and the deteriorated group). Respite stay in a nursing home service use and more types of service use were related to experiencing a deterioration of care needs level in lower care needs level community-dwelling elderly persons in Japan. Further, medical management by a physician service was related to experiencing a deterioration of care needs level in higher care needs level community-dwelling elderly persons. PMID- 20025750 TI - Cognitive impairment in patients with fibromyalgia syndrome as assessed by the mini-mental state examination. AB - BACKGROUND: This study evaluated the frequency of cognitive impairment in patients with Fibromyalgia syndrome (FMS) using the Mini Mental State Examination (MMSE). METHODS: We analyzed baseline data from all 46 patients with FMS and 92 age- and sex-matched controls per diagnosis of neuropathic (NeP) or mixed pain (MP) selected from a larger prospective study. RESULTS: FMS had a slight but statistically significant lower score in the adjusted MMSE score (26.9; 95% CI 26.7-27.1) than either NeP (27.3; 95% CI 27.2-27.4) or MP (27.3; 27.2-27.5). The percentage of patients with congnitive impairment (adjusted MMSE or= II. METHODS: 118 patients (median age 64 years; male : female ratio 2.5 : 1) with pathological proven rectal cancer (clinical stage II 50%, III 41.5%, IV 8.5%) were treated preoperatively with twice daily radiotherapy of 2.9 Gy single fraction dose to a total dose of 29 Gy; surgery was performed immediately in the following week with total mesorectal excision (TME). Adjuvant 5-FU based chemotherapy was planned for pathological stage UICC >or= II. RESULTS: After low anterior resection (70%) and abdominoperineal resection (30%), pathology showed stage UICC I (27.1%), II (25.4%), III (37.3%) and IV (9.3%). Perioperative mortality was 3.4% and perioperative complications were observed in 22.8% of the patients. Adjuvant chemotherapy was given in 75.3% of patients with pathological stage UICC >or= II. After median follow-up of 46 months, five-year overall survival was 67%, cancer-specific survival 76%, local control 92% and freedom from systemic progression 75%. Late toxicity > grade II was observed in 11% of the patients. CONCLUSIONS: Preoperative short-course radiotherapy, total mesorectal excision and adjuvant chemotherapy for pathological stage UICC >or= II achieved excellent local control and favorable survival. PMID- 20025753 TI - Evolution of surface-based deformable image registration for adaptive radiotherapy of non-small cell lung cancer (NSCLC). AB - BACKGROUND: To evaluate the performance of surface-based deformable image registration (DR) for adaptive radiotherapy of non-small cell lung cancer (NSCLC). METHODS: Based on 13 patients with locally advanced NSCLC, CT images acquired at treatment planning, midway and the end of the radio- (n = 1) or radiochemotherapy (n = 12) course were used for evaluation of DR. All CT images were manually [gross tumor volume (GTV)] and automatically [organs-at-risk (OAR) lung, spinal cord, vertebral spine, trachea, aorta, outline] segmented. Contours were transformed into 3D meshes using the Pinnacle treatment planning system and corresponding mesh points defined control points for DR with interpolation within the structures. Using these deformation maps, follow-up CT images were transformed into the planning images and compared with the original planning CT images. RESULTS: A progressive tumor shrinkage was observed with median GTV volumes of 170 cm(3) (range 42 cm(3) - 353 cm(3)), 124 cm(3) (19 cm(3) - 325 cm(3)) and 100 cm(3) (10 cm(3) - 270 cm(3)) at treatment planning, mid-way and at the end of treatment. Without DR, correlation coefficients (CC) were 0.76 +/- 0.11 and 0.74 +/- 0.10 for comparison of the planning CT and the CT images acquired mid-way and at the end of treatment, respectively; DR significantly improved the CC to 0.88 +/- 0.03 and 0.86 +/- 0.05 (p = 0.001), respectively. With manual landmark registration as reference, DR reduced uncertainties on the GTV surface from 11.8 mm +/- 5.1 mm to 2.9 mm +/- 1.2 mm. Regarding the carina and intrapulmonary vessel bifurcations, DR reduced uncertainties by about 40% with residual errors of 4 mm to 6 mm on average. Severe deformation artefacts were observed in patients with resolving atelectasis and pleural effusion, in one patient, where the tumor was located around large bronchi and separate segmentation of the GTV and OARs was not possible, and in one patient, where no clear shrinkage but more a decay of the tumor was observed. DISCUSSION: The surface-based DR performed accurately for the majority of the patients with locally advanced NSCLC. However, morphological response patterns were identified, where results of the surface-based DR are uncertain. PMID- 20025754 TI - Measurement properties of the Dizziness Handicap Inventory by cross-sectional and longitudinal designs. AB - BACKGROUND: The impact of dizziness on quality of life is often assessed by the Dizziness Handicap Inventory (DHI), which is used as a discriminate and evaluative measure. The aim of the present study was to examine reliability and validity of a translated Norwegian version (DHI-N), also examining responsiveness to important change in the construct being measured. METHODS: Two samples (n = 92 and n = 27) included participants with dizziness of mainly vestibular origin. A cross-sectional design was used to examine the factor structure (exploratory factor analysis), internal consistency (Cronbach's alpha), concurrent validity (Pearson's product moment correlation r), and discriminate ability (ROC curve analysis). Longitudinal designs were used to examine test-retest reliability (intraclass correlation coefficient (ICC) statistics, smallest detectable difference (SDD)), and responsiveness (Pearson's product moment correlation, ROC curve analysis; area under the ROC curve (AUC), and minimally important change (MIC)). The DHI scores range from 0 to 100. RESULTS: Factor analysis revealed a different factor structure than the original DHI, resulting in dismissal of subscale scores in the DHI-N. Acceptable internal consistency was found for the total scale (alpha = 0.95). Concurrent correlations between the DHI-N and other related measures were moderate to high, highest with Vertigo Symptom Scale-short form-Norwegian version (r = 0.69), and lowest with preferred gait (r = - 0.36). The DHI-N demonstrated excellent ability to discriminate between participants with and without 'disability', AUC being 0.89 and best cut-off point = 29 points. Satisfactory test-retest reliability was demonstrated, and the change for an individual should be >/= 20 DHI-N points to exceed measurement error (SDD). Correlations between change scores of DHI-N and other self-report measures of functional health and symptoms were high (r = 0.50 - 0.57). Responsiveness of the DHI-N was excellent, AUC = 0.83, discriminating between self-perceived 'improved' versus 'unchanged' participants. The MIC was identified as 11 DHI-N points. CONCLUSIONS: The DHI-N total scale demonstrated satisfactory measurement properties. This is the first study that has addressed and demonstrated responsiveness to important change of the DHI, and provided values of SDD and MIC to help interpret change scores. PMID- 20025756 TI - In silico evaluation of predicted regulatory interactions in Arabidopsis thaliana. AB - BACKGROUND: Prediction of transcriptional regulatory mechanisms in Arabidopsis has become increasingly critical with the explosion of genomic data now available for both gene expression and gene sequence composition. We have shown in previous work 1, that a combination of correlation measurements and cis-regulatory element (CRE) detection methods are effective in predicting targets for candidate transcription factors for specific case studies which were validated. However, to date there has been no quantitative assessment as to which correlation measures or CRE detection methods used alone or in combination are most effective in predicting TF-->target relationships on a genome-wide scale. RESULTS: We tested several widely used methods, based on correlation (Pearson and Spearman Rank correlation) and cis-regulatory element (CRE) detection (>or=1 CRE or CRE over representation), to determine which of these methods individually or in combination is the most effective by various measures for making regulatory predictions. To predict the regulatory targets of a transcription factor (TF) of interest, we applied these methods to microarray expression data for genes that were regulated over treatment and control conditions in wild type (WT) plants. Because the chosen data sets included identical experimental conditions used on TF over-expressor or T-DNA knockout plants, we were able to test the TF-->target predictions made using microarray data from WT plants, with microarray data from mutant/transgenic plants. For each method, or combination of methods, we computed sensitivity, specificity, positive and negative predictive value and the F measure of balance between sensitivity and positive predictive value (precision). This analysis revealed that the >or=1 CRE and Spearman correlation (used alone or in combination) were the most balanced CRE detection and correlation methods, respectively with regard to their power to accurately predict regulatory-target interactions. CONCLUSION: These findings provide an approach and guidance for researchers interested in predicting transcriptional regulatory mechanisms using microarray data that they generate (or microarray data that is publically available) combined with CRE detection in promoter sequence data. PMID- 20025755 TI - Fine particulate air pollution and its components in association with cause specific emergency admissions. AB - BACKGROUND: Although the association between exposure to particulate matter and health is well established, there remains uncertainty as to whether certain chemical components are more harmful than others. We explored whether the association between cause-specific hospital admissions and PM(2.5) was modified by PM(2.5) chemical composition. METHODS: We estimated the association between daily PM(2.5) and emergency hospital admissions for cardiac causes (CVD), myocardial infarction (MI), congestive heart failure (CHF), respiratory disease, and diabetes in 26 US communities, for the years 2000-2003. Using meta regression, we examined how this association was modified by season- and community-specific PM(2.5) composition, controlling for seasonal temperature as a surrogate for ventilation. RESULTS: For a 10 microg/m3 increase in 2-day averaged PM(2.5) concentration we found an increase of 1.89% (95% CI: 1.34- 2.45) in CVD, 2.25% (95% CI: 1.10- 3.42) in MI, 1.85% (95% CI: 1.19- 2.51) in CHF, 2.74% (95% CI: 1.30- 4.2) in diabetes, and 2.07% (95% CI: 1.20- 2.95) in respiratory admissions. The association between PM2.5 and CVD admissions was significantly modified when the mass was high in Br, Cr, Ni, and Na(+), while mass high in As, Cr, Mn, OC, Ni, and Na(+) modified MI, and mass high in As, OC, and SO(4)(2-) modified diabetes admissions. For these species, an interquartile range increase in their relative proportion was associated with a 1-2% additional increase in daily admissions per 10 microg/m(3) increase in mass. CONCLUSIONS: We found that PM(2.5) mass higher in Ni, As, and Cr, as well as Br and OC significantly increased its effect on hospital admissions. This result suggests that particles from industrial combustion sources and traffic may, on average, have greater toxicity. PMID- 20025757 TI - Use of facility assessment data to improve reproductive health service delivery in the Democratic Republic of the Congo. AB - BACKGROUND: Prolonged exposure to war has severely impacted the provision of health services in the Democratic Republic of the Congo (DRC). Health infrastructure has been destroyed, health workers have fled and government support to health care services has been made difficult by ongoing conflict. Poor reproductive health (RH) indicators illustrate the effect that the prolonged crisis in DRC has had on the on the reproductive health (RH) of Congolese women. In 2007, with support from the RAISE Initiative, the International Rescue Committee (IRC) and CARE conducted baseline assessments of public hospitals to evaluate their capacities to meet the RH needs of the local populations and to determine availability, utilization and quality of RH services including emergency obstetric care (EmOC) and family planning (FP). METHODS: Data were collected from facility assessments at nine general referral hospitals in five provinces in the DRC during March, April and November 2007. Interviews, observation and clinical record review were used to assess the general infrastructure, EmOC and FP services provided, and the infection prevention environment in each of the facilities. RESULTS: None of the nine hospitals met the criteria for classification as an EmOC facility (either basic or comprehensive). Most facilities lacked any FP services. Shortage of trained staff, essential supplies and medicines and poor infection prevention practices were consistently documented. All facilities had poor systems for routine monitoring of RH services, especially with regard to EmOC. CONCLUSIONS: Women's lives can be saved and their well-being improved with functioning RH services. As the DRC stabilizes, IRC and CARE in partnership with the local Ministry of Health and other service provision partners are improving RH services by: 1) providing necessary equipment and renovations to health facilities; 2) improving supply management systems; 3) providing comprehensive competency-based training for health providers in RH and infection prevention; 4) improving referral systems to the hospitals; 5) advocating for changes in national RH policies and protocols; and 6) providing technical assistance for monitoring and evaluation of key RH indicators. Together, these initiatives will improve the quality and accessibility of RH services in the DRC - services which are urgently needed and to which Congolese women are entitled by international human rights law. PMID- 20025758 TI - Early recognition of heart failure in patients with diabetes type 2 in primary care. A prospective diagnostic efficiency study. (UHFO-DM2). AB - BACKGROUND: We hypothesize that the prevalence of unknown heart failure in diabetic patients aged 60 years and over is relatively high (15% or more) and that a cost-effective strategy can be developed to detect heart failure in these patients. The strategy is expected to include some signs and symptoms (such as dyspnoea, orthopnoea, pulmonary crepitations and laterally displaced apical beat), natriuretic peptide measurements (Amino-terminal B-type natriuretic peptide) and possibly electrocardiography. In a subset of patients straightforward echocardiography may show to be cost-effective. With information from our study the detection of previously unknown heart failure in diabetic patients could be improved and enable the physician to initiate beneficial morbidity and mortality reducing heart failure treatment more timely. PRIMARY OBJECTIVES: - To assess the prevalence of (previously unrecognised) heart failure in primary care patients with diabetes type 2.- To establish the most cost effective diagnostic strategy to detect unrecognised heart failure in these patients. SECONDARY OBJECTIVES: - To assess the impact of heart failure, and the combination of a new diagnosis with accordingly treatment in patients with diabetes type 2 on health status. METHODS/DESIGN: DESIGN: A prospective diagnostic efficiency study. PATIENT POPULATION: Patients aged 60 years and older with diabetes type 2 from primary care, enlisted with the diabetes service of the Diagnostic Center in Etten-Leur (SHL)All participants will be investigated at the cardiology out-patient department of the regional hospital (Oosterschelde Hospital in Goes, Zeeland, the Netherlands) during a single 1.5 hour standardised diagnostic assessment, including history taking, physical examination, electrocardiography, echocardiography, blood tests, and Health status questionnaires. Patients will be asked if we can contact them afterwards for follow-up and for repeating the questionnaires after three and 12 months.Main study parameters/endpoints: Prevalence (with exact 95% confidence intervals) of (previously unrecognised) heart failure (systolic and 'isolated' diastolic) and the diagnostic value of signs and symptoms, NT-proBNP, electrocardiography and a combination of these items. The cost-effectiveness of different diagnostic strategies. Impact of heart failure and the combination of a new diagnosis with accordingly treatment on health status. TRIAL REGISTRATION: CCMO register NL2271704108. PMID- 20025759 TI - Use of supplementary phenotype to identify additional rheumatoid arthritis loci in a linkage analysis of 342 UK affected sibling pair families. AB - BACKGROUND: Although rheumatoid arthritis has been shown to have moderately strong genetic component, both linked loci identified in linkage analyses and susceptibility variants from association studies are short of adequately accounting for a comprehensive catalogue of the molecular factors underlying this complex disease. The objective of this study was to use supplementary phenotype based on cumulative hazard of rheumatoid arthritis to identify linkage evidence for new and additional rheumatoid arthritis loci in a genome-wide linkage analysis of 342 affected sibling pair families from the United Kingdom. METHODS: Using proportional hazards model, we estimated cumulative hazard of rheumatoid arthritis and then used it as a quantitative trait in a non-parametric multipoint variance component linkage analysis with 353 microsatellite markers distributed across the 22 autosomal chromosomes. RESULTS: We identified 3 new loci with genome-wide suggestive linkage evidence for rheumatoid arthritis on 9q21.13, 15p11.1 and 20q13.33. Our results also confirmed previously reported linkage evidence in the HLA-DRB1 region on chromosome 6 and on locus 1q32.1. CONCLUSION: This study demonstrates the potential for information gain through the use of supplementary phenotypes in genetic study of complex diseases to identify new and additional potential linked loci that are not detected by linkage analysis of traditional phenotypes; and our results provide further evidence of the involvement of multiple loci in the genetic aetiology of rheumatoid arthritis. PMID- 20025760 TI - Duration of red blood cell storage and outcomes in pediatric cardiac surgery: an association found for pump prime blood. AB - INTRODUCTION: Cardiac surgery using cardiopulmonary bypass in newborns, infants and small children often requires intraoperative red blood cell transfusions to prime the circuit and oxygenator and to replace blood lost during surgery. The purpose of this study was to investigate the influence of red blood cell storage time prior to transfusion on postoperative morbidity in pediatric cardiac operations. METHODS: One hundred ninety-two consecutive children aged five years or less who underwent cardiac operations using cardiopulmonary bypass and who received red blood cells for priming the cardiopulmonary bypass circuit comprised the blood-prime group. Forty-seven patients receiving red blood cell transfusions after cardiopulmonary bypass were separately analyzed. Patients in the blood prime group were divided into two groups based on the duration of storage of the red blood cells they received. The newer blood group included patients who received only red blood cells stored for less than or equal to four days and the older blood group included patients who received red blood cells stored for more than four days. RESULTS: Patients in the newer blood group had a significantly lower rate of pulmonary complications (3.5% versus 14.4%; P = 0.011) as well as a lower rate of acute renal failure (0.8% versus 5.2%; P = 0.154) than patients in the older blood group. Major complications (calculated as a composite score based on pulmonary, neurological, and gastroenterological complications, sepsis and acute renal failure) were found in 6.9% of the patients receiving newer blood and 17.1% of the patients receiving older blood (P = 0.027). After adjusting for other possible confounding variables, red blood cell storage time remained an independent predictor of major morbidity. The same association was not found for patients receiving red blood cell transfusions after cardiopulmonary bypass. CONCLUSIONS: The storage time of the red blood cells used for priming the cardiopulmonary bypass circuit in cardiac operations on newborns and young infants is an independent risk factor for major postoperative morbidity. Pulmonary complications, acute renal failure, and infections are the main complications associated with increased red blood cell storage time. PMID- 20025761 TI - The prevalence of neovascularity in patients clinically diagnosed with rotator cuff tendinopathy. AB - BACKGROUND: Shoulder dysfunction is common and pathology of the rotator cuff tendons and subacromial bursa are considered to be a major cause of pain and morbidity. Although many hypotheses exist there is no definitive understanding as to the origin of the pain arising from these structures. Research investigations from other tendons have placed intra-tendinous neovascularity as a potential mechanism of pain production. The prevalence of neovascularity in patients with a clinical diagnosis of rotator cuff tendinopathy is unknown. As such the primary aim of this pilot study was to investigate if neovascularity could be identified and to determine the prevalence of neovascularity in the rotator cuff tendons and subacromial bursa in subjects with unilateral shoulder pain clinically assessed to be rotator cuff tendinopathy. The secondary aims were to investigate the association between the presence of neovascularity and pain, duration of symptoms, and, neovascularity and shoulder function. METHODS: Patients with a clinical diagnosis of unilateral rotator cuff tendinopathy referred for a routine diagnostic ultrasound (US) scan in a major London teaching hospital formed the study population. At referral patients were provided with an information document. On the day of the scan (on average, at least one week later) the patients agreeing to participate were taken through the consent process and underwent an additional clinical examination prior to undergoing a bilateral grey scale and colour Doppler US examination (symptomatic and asymptomatic shoulder) using a Philips HDI 5000 Sono CT US machine. The ultrasound scans were performed by one of two radiologists who recorded their findings and the final assessment was made by a third radiologist blinded both to the clinical examination and the ultrasound examination. The findings of the radiologists who performed the scans and the blinded radiologist were compared and any disagreements were resolved by consensus. RESULTS: Twenty-six patients agreed to participate and formed the study population. Of these, 6 subjects were not included in the final assessment following the pre-scan clinical investigation. This is because one subject had complete cessation of symptoms between the time of the referral and entry into the trial. Another five had developed bilateral shoulder pain during the same period. The mean age of the 20 subjects forming the study population was 50.2 (range 32-69) years (SD = 10.9) and the mean duration of symptoms was 22.6 (range .75 to 132) months (SD = 40.1). Of the 20 subjects included in the formal analysis, 13 subjects (65%) demonstrated neovascularity in the symptomatic shoulder and 5 subjects (25%) demonstrated neovascularity in the asymptomatic shoulder. The subject withdrawn due to complete cessation of symptoms was not found to have neovascularity in either shoulder and of the 5 withdrawn due to bilateral symptoms; two subjects were found to have signs of bilateral neovascularity, one subject demonstrated neovascularity in one shoulder and two subjects in neither shoulder. CONCLUSIONS: This study demonstrated that neovascularity does occur in subjects with a clinical diagnosis of rotator cuff tendinopathy and to a lesser extent in asymptomatic shoulders. In addition, the findings of this investigation did not identify an association between the presence of neovascularity; and pain, duration of symptoms or shoulder function. Future research is required to determine the relevance of these findings. PMID- 20025762 TI - Evaluation of a new disposable silicon limbal relaxing incision knife by experienced users. AB - BACKGROUND: Previous research has suggested that the silicon BD Atomic Edge knife has superior performance characteristics when compared to a metal knife and performance similar to diamond knife when making various incisions. This study was designed to determine whether a silicon accurate depth knife has equivalent performance characteristics when compared to a diamond limbal relaxing incision (LRI) knife and superior performance characteristics when compared to a steel accurate depth knife when creating limbal relaxing incision. METHODS: Sixty-five ophthalmic surgeons with limbal relaxing incision experience created limbal relaxing incisions in ex-vivo porcine eyes with silicon and steel accurate depth knives and diamond LRI knives. The ophthalmic surgeons rated multiple performance characteristics of the knives on Visual Analog Scales. RESULTS: The observed differences between the silicon knife and diamond knife were found to be insignificant. The mean ratio between the performance of the silicon knife and the diamond knife was shown to be greater than 90% (with 95% confidence). The silicon knife's mean performance was significantly higher than the performance of the steel knife for all characteristics. (p-value < .05) CONCLUSIONS: For experienced users, the silicon accurate depth knife was found to be equivalent in performance to the diamond LRI knife and superior to the steel accurate depth knife when making limbal relaxing incisions in ex vivo porcine eyes. Disposable silicon LRI knives may be an alternative to diamond LRI knives. PMID- 20025763 TI - Model system for the analysis of cell surface expression of human ABCA1. AB - BACKGROUND: The ABCA1 protein plays a pivotal role in reverse cholesterol transport, by mediating the generation of HDL particles and removing cellular cholesterol. Both the proper expression of ABCA1 in the plasma membrane and the internalization along with apoA-I are required for function. Therefore, we developed a model system to investigate the effect of clinically relevant drugs on the cell surface appearance of ABCA1. RESULTS: By retroviral transduction system, we established stable mammalian cell lines expressing functional and non functional ABCA1 variants, tagged with an extracellular hemagglutinin epitope. After characterization of the expression, proper localization and function of different ABCA1 variants, we followed quantitatively their cell surface expression by immunofluorescent staining, using flow cytometry. As expected, we found increased cell surface expression of ABCA1 after treatment with a calpain inhibitor, and observed a strong decrease in plasma membrane ABCA1 expression upon treatment with a trans-Golgi transport inhibitor, Brefeldin A. We tested cholesterol level lowering drugs and other potential inhibitors of ABCA1. Here we demonstrate that ezetimibe affects ABCA1 cell surface expression only in the case of a functional ABCA1. CONCLUSIONS: Our model system allows a quantitative detection of cell surface expression of ABCA1, screening of substrates or specific inhibitors, and investigating transport regulation. PMID- 20025764 TI - Comparative proteomic analysis of pathogenic and non-pathogenic strains from the swine pathogen Mycoplasma hyopneumoniae. AB - BACKGROUND: Mycoplasma hyopneumoniae is a highly infectious swine pathogen and is the causative agent of enzootic pneumonia (EP). Following the previous report of a proteomic survey of the pathogenic 7448 strain of swine pathogen, Mycoplasma hyopneumoniae, we performed comparative protein profiling of three M. hyopneumoniae strains, namely the non-pathogenic J strain and the two pathogenic strains 7448 and 7422. RESULTS: In 2DE comparisons, we were able to identify differences in expression levels for 67 proteins, including the overexpression of some cytoadherence-related proteins only in the pathogenic strains. 2DE immunoblot analyses allowed the identification of differential proteolytic cleavage patterns of the P97 adhesin in the three strains. For more comprehensive protein profiling, an LC-MS/MS strategy was used. Overall, 35% of the M. hyopneumoniae genome coding capacity was covered. Partially overlapping profiles of identified proteins were observed in the strains with 81 proteins identified only in one strain and 54 proteins identified in two strains. Abundance analysis of proteins detected in more than one strain demonstrates the relative overexpression of 64 proteins, including the P97 adhesin in the pathogenic strains. CONCLUSIONS: Our results indicate the physiological differences between the non-pathogenic strain, with its non-infective proliferate lifestyle, and the pathogenic strains, with its constitutive expression of adhesins, which would render the bacterium competent for adhesion and infection prior to host contact. PMID- 20025766 TI - Spectrum of abdominal organ injury in a primary blast type. AB - INTRODUCTION: Abdominal organ injury in a primary blast type is always challenging for diagnosis. Air containing abdominal viscera is most vulnerable to effects of primary blast injury. In any patient exposed to a primary blast wave who presents with an acute abdomen, an abdominal organ injury is to be kept in a clinical suspicion. AIM: Study various abdominal organ injuries occurring in a primary type of blast injury. MATERIAL AND METHODS: All those who had exploratory laparotomy for abdominal organ injuries after a primary blast injury for a period of 10 years from January 1998 - January 2008 were included in this retrospective study. RESULTS: Total 154 patients had laparotomy for abdominal organ injuries with a primary blast type of injury. Small intestine was damaged in 48 patients (31.1%) followed by spleen in 22.7% cases. 54 patients (35.06%) had more than one organ injured. Liver laceration was present in 30 patients (19.48%). Multiple small gut perforations were present in 37 patients (77.08%). Negative laparotomy was found in 5 patients (3.24%) whereas 3 (1.94%) had re-exploration. Mortality was present in 6 patients (3.89%). CONCLUSIONS: Primary blast injury causes varied abdominal organ injuries. Single or multiple organ damage can be there. Small intestine is commonest viscera injured. Laparotomy gives final diagnosis. PMID- 20025765 TI - Differential involvement of trigeminal transition zone and laminated subnucleus caudalis in orofacial deep and cutaneous hyperalgesia: the effects of interleukin 10 and glial inhibitors. AB - BACKGROUND: In addition to caudal subnucleus caudalis (Vc) of the spinal trigeminal complex, recent studies indicate that the subnuclei interpolaris/caudalis (Vi/Vc) transition zone plays a unique role in processing deep orofacial nociceptive input. Studies also suggest that glia and inflammatory cytokines contribute to the development of persistent pain. By systematically comparing the effects of microinjection of the antiinflammatory cytokine interleukin (IL)-10 and two glial inhibitors, fluorocitrate and minocycline, we tested the hypothesis that there was a differential involvement of Vi/Vc and caudal Vc structures in deep and cutaneous orofacial pain. RESULTS: Deep or cutaneous inflammatory hyperalgesia, assessed with von Frey filaments, was induced in rats by injecting complete Freund's adjuvant (CFA) into the masseter muscle or skin overlying the masseter, respectively. A unilateral injection of CFA into the masseter or skin induced ipsilateral hyperalgesia that started at 30 min, peaked at 1 d and lasted for 1-2 weeks. Secondary hyperalgesia on the contralateral site also developed in masseter-, but not skin-inflamed rats. Focal microinjection of IL-10 (0.006-1 ng), fluorocitrate (1 microg), and minocycline (0.1-1 microg) into the ventral Vi/Vc significantly attenuated masseter hyperalgesia bilaterally but without an effect on hyperalgesia after cutaneous inflammation. Injection of the same doses of these agents into the caudal Vc attenuated ipsilateral hyperalgesia after masseter and skin inflammation, but had no effect on contralateral hyperalgesia after masseter inflammation. Injection of CFA into the masseter produced significant increases in N-methyl-D-aspartate (NMDA) receptor NR1 serine 896 phosphorylation and glial fibrillary acidic protein (GFAP) levels, a marker of reactive astrocytes, in Vi/Vc and caudal Vc. In contrast, cutaneous inflammation only produced similar increases in the Vc. CONCLUSION: These results support the hypothesis that the Vi/Vc transition zone is involved in deep orofacial injury and suggest that glial inhibition and interruption of the cytokine cascade after inflammation may provide pain relief. PMID- 20025767 TI - Investigating AP-2 and YY1 protein expression as a cause of high HER2 gene transcription in breast cancers with discordant HER2 gene amplification. AB - INTRODUCTION: Candidacy for anti-HER2 adjuvant therapy in breast cancer is assessed using tumour HER2 status but recently it has been proposed that the transcription factors AP-2alpha and YY1 may cause Her2 protein overexpression independently of gene amplification. METHODS: We characterised AP-2alpha/beta, AP 2alpha and YY1 with HER2 gene and protein expression, other relevant biomarkers, and clinical outcome using tissue microarrays (TMAs) and immunohistochemistry in a large (n = 1,176) clinically annotated series of early stage operable breast cancer. The associations and prognostic independence of AP-2 and YY1 was assessed in all patients and an oestrogen receptor negative subgroup. RESULTS: Nuclear expression of AP-2alpha/beta, AP-2alpha and YY1 was detected in 23%, 44% and 33% of cases respectively. AP-2alpha/beta significantly correlated with YY1 and both markers were increased in luminal oestrogen receptor (ER) positive tumours of small size and low grade but only AP-2alpha/beta correlated with good prognosis breast cancer specific survival and disease free interval (BCSS and DFI). These characteristics were lost in oestrogen receptor negative patients. AP-2alpha also correlated with luminal-type tumours but not with YY1 expression or good prognosis. AP-2alpha and YY1 showed a significant correlation with Her2 protein expression and in addition, YY1 correlated with HER2 gene expression. Discordant HER2 gene and protein expression was identified in six cases (0.71% of the study group) with four of these showing AP-2alpha but absence of AP-2alpha/beta and YY1 expression. CONCLUSIONS: AP-2alpha/beta and YY1 are markers of good prognosis principally due to their association with oestrogen receptor but are not independent predictors. Discordant HER2 protein/gene expression is a rare event that is not always explained by the actions of AP-2 and YY1. PMID- 20025768 TI - PreDisorder: ab initio sequence-based prediction of protein disordered regions. AB - BACKGROUND: Disordered regions are segments of the protein chain which do not adopt stable structures. Such segments are often of interest because they have a close relationship with protein expression and functionality. As such, protein disorder prediction is important for protein structure prediction, structure determination and function annotation. RESULTS: This paper presents our protein disorder prediction server, PreDisorder. It is based on our ab initio prediction method (MULTICOM-CMFR) which, along with our meta (or consensus) prediction method (MULTICOM), was recently ranked among the top disorder predictors in the eighth edition of the Critical Assessment of Techniques for Protein Structure Prediction (CASP8). We systematically benchmarked PreDisorder along with 26 other protein disorder predictors on the CASP8 data set and assessed its accuracy using a number of measures. The results show that it compared favourably with other ab initio methods and its performance is comparable to that of the best meta and clustering methods. CONCLUSION: PreDisorder is a fast and reliable server which can be used to predict protein disordered regions on genomic scale. It is available at http://casp.rnet.missouri.edu/predisorder.html. PMID- 20025769 TI - Economic and other barriers to adopting recommendations to prevent childhood obesity: results of a focus group study with parents. AB - BACKGROUND: Parents are integral to the implementation of obesity prevention and management recommendations for children. Exploration of barriers to and facilitators of parental decisions to adopt obesity prevention recommendations will inform future efforts to reduce childhood obesity. METHODS: We conducted 4 focus groups (2 English, 2 Spanish) among a total of 19 parents of overweight (BMI >or= 85th percentile) children aged 5-17 years. The main discussion focused on 7 common obesity prevention recommendations: reducing television (TV) watching, removing TV from child's bedroom, increasing physically active games, participating in community or school-based athletics, walking to school, walking more in general, and eating less fast food. Parents were asked to discuss what factors would make each recommendation more difficult (barriers) or easier (facilitators) to follow. Participants were also asked about the relative importance of economic (time and dollar costs/savings) barriers and facilitators if these were not brought into the discussion unprompted. RESULTS: Parents identified many barriers but few facilitators to adopting obesity prevention recommendations for their children. Members of all groups identified economic barriers (time and dollar costs) among a variety of pertinent barriers, although the discussion of dollar costs often required prompting. Parents cited other barriers including child preference, difficulty with changing habits, lack of information, lack of transportation, difficulty with monitoring child behavior, need for assistance from family members, parity with other family members, and neighborhood walking safety. Facilitators identified included access to physical activity programs, availability of alternatives to fast food and TV which are acceptable to the child, enlisting outside support, dietary information, involving the child, setting limits, making behavior changes gradually, and parental change in shopping behaviors and own eating behaviors. CONCLUSIONS: Parents identify numerous barriers to adopting obesity prevention recommendations, most notably child and family preferences and resistance to change, but also economic barriers. Intervention programs should consider the context of family priorities and how to overcome barriers and make use of relevant facilitators during program development. PMID- 20025770 TI - Continuity of care for children with complex chronic health conditions: parents' perspectives. AB - BACKGROUND: Continuity of care has been explored largely from academic and service provider perspectives, and in relation to adult patient/client groups. We interviewed parents of children with complex chronic health conditions to examine how their experiences and perceptions of continuity of care fit with these perspectives; and to identify the salient factors in the experience of, and factors contributing to, continuity in this population. METHODS: Parents of 47 elementary school-aged children with spina bifida, Down syndrome, attention deficit/hyperactivity disorder, Duchenne muscular dystrophy or cystic fibrosis participated in semi-structured interviews. Parents described and mapped the pattern of their interactions with service providers over time in all domains relevant to their child's health, well-being, and development (medical, rehabilitational, educational, and social supportive services), with particular attention paid to their perceptions of connectedness or coherency in these interactions. Verbatim transcripts were analyzed thematically using a framework approach to impose structure regarding parents' perspectives on continuity of care. RESULTS: Existing academic concepts of relational, informational and management continuity were all discernable in parents' narratives. A thorough knowledge of the child on the part of service providers emerged as extremely important to parents; such knowledge was underpinned by continuity of personal relationships, principally, and also by written information. For this population, notions of continuity extend to the full range of service providers these children and families need to achieve optimal health status, and are not limited to physicians and nurses. Communication among providers was seen as integral to perceived continuity. Compartmentalization of services and information led to parents assuming a necessary, though at times, uncomfortable, coordinating role. Geographic factors, institutional structures and practices, provider attitudes, and, on occasion, parent preferences and judgments, were all found to create barriers to "seamless" management and provision of care continuity across providers, settings, and sectors. CONCLUSIONS: These findings add new perspectives to the understanding of continuity within chronically ill children's health care. They are relevant to contemporary initiatives to improve continuity of services to children with special health care needs, demonstrate the need for parental support of their important role in maintaining continuity, and suggest avenues for further research. PMID- 20025771 TI - The commonly-used DNA probe for diffusely-adherent Escherichia coli cross-reacts with a subset of enteroaggregative E. coli. AB - BACKGROUND: The roles of diffusely-adherent Escherichia coli (DAEC) and enteroaggregative E. coli (EAEC) in disease are not well understood, in part because of the limitations of diagnostic tests for each of these categories of diarrhoea-causing E. coli. A HEp-2 adherence assay is the Gold Standard for detecting both EAEC and DAEC but DNA probes with limited sensitivity are also employed. RESULTS: We demonstrate that the daaC probe, conventionally used to detect DAEC, cross-reacts with a subset of strains belonging to the EAEC category. The cross hybridization is due to 84% identity, at the nucleotide level, between the daaC locus and the aggregative adherence fimbriae II cluster gene, aafC, present in some EAEC strains. Because aaf-positive EAEC show a better association with diarrhoea than other EAEC, this specific cross-hybridization may have contributed to an over-estimation of the association of daaC with disease in some studies. We have developed a discriminatory PCR-RFLP protocol to delineate EAEC strains detected by the daaC probe in molecular epidemiological studies. CONCLUSIONS: A PCR-RFLP protocol described herein can be used to identify aaf positive EAEC and daaC-positive DAEC and to delineate these two types of diarrhoeagenic E. coli, which both react with the daaC probe. This should help to improve current understanding and future investigations of DAEC and EAEC epidemiology. PMID- 20025772 TI - Diagnostic accuracy of pattern differentiation algorithm based on Chinese medicine theory: a stochastic simulation study. AB - BACKGROUND: Clinical practice of Chinese medicine requires little information for differentiation of Zang-fu patterns. This study is to test the impact of information amount on the diagnostic accuracy of pattern differentiation algorithm (PDA) using stochastic simulation of cases. METHODS: A dataset with 69 Zang-fu single patterns was used with manifestations according to the Four Examinations, namely inspection (Ip), auscultation and olfaction (AO), inquiry (Iq) and palpation (P). A variable quantity of available information (N(%)) was randomly sampled to generate 100 true positive and 100 true negative manifestation profiles per pattern to which PDA was applied. Four runs of simulations were used according to the Four Examinations: Ip, Ip+AO, Ip+AO+Iq and Ip+AO+Iq+P. The algorithm performed pattern differentiation by ranking a list of diagnostic hypotheses by the amount of explained information F(%). Accuracy, sensitivity, specificity and negative and positive predictive values were calculated. RESULTS: Use the Four Examinations resulted in the best accuracy with the smallest cutoff value (N(%)= 28.5%), followed by Ip+AO+Iq (33.5%), Ip+AO (51.5%) and Ip (52.0%). All tested combinations provided concave-shaped curves for accuracy, indicating an optimal value subject to N(%-cutoff). Use of N(% cutoff) as a secondary criterion resulted in 94.7% (94.3; 95.1) accuracy, 89.8% (89.1; 90.6) sensitivity, and 99.5% (99.3; 99.7) specificity with the Four Examinations. CONCLUSION: Pattern differentiation based on both explained and optimum available information (F(%) and N(%-cutoff)) is more accurate than using explained and available information without cutoff (F(%) and N(%)). Both F(%) and N(%-cutoff) should be used as PDA's objective criteria to perform Zang-fu single pattern differentiation. PMID- 20025773 TI - Optimal classifier selection and negative bias in error rate estimation: an empirical study on high-dimensional prediction. AB - BACKGROUND: In biometric practice, researchers often apply a large number of different methods in a "trial-and-error" strategy to get as much as possible out of their data and, due to publication pressure or pressure from the consulting customer, present only the most favorable results. This strategy may induce a substantial optimistic bias in prediction error estimation, which is quantitatively assessed in the present manuscript. The focus of our work is on class prediction based on high-dimensional data (e.g. microarray data), since such analyses are particularly exposed to this kind of bias. METHODS: In our study we consider a total of 124 variants of classifiers (possibly including variable selection or tuning steps) within a cross-validation evaluation scheme. The classifiers are applied to original and modified real microarray data sets, some of which are obtained by randomly permuting the class labels to mimic non informative predictors while preserving their correlation structure. RESULTS: We assess the minimal misclassification rate over the different variants of classifiers in order to quantify the bias arising when the optimal classifier is selected a posteriori in a data-driven manner. The bias resulting from the parameter tuning (including gene selection parameters as a special case) and the bias resulting from the choice of the classification method are examined both separately and jointly. CONCLUSIONS: The median minimal error rate over the investigated classifiers was as low as 31% and 41% based on permuted uninformative predictors from studies on colon cancer and prostate cancer, respectively. We conclude that the strategy to present only the optimal result is not acceptable because it yields a substantial bias in error rate estimation, and suggest alternative approaches for properly reporting classification accuracy. PMID- 20025774 TI - Developmental dynamics of cone photoreceptors in the eel. AB - BACKGROUND: Many fish alter their expressed visual pigments during development. The number of retinal opsins expressed and their type is normally related to the environment in which they live. Eels are known to change the expression of their rod opsins as they mature, but might they also change the expression of their cone opsins? RESULTS: The Rh2 and Sws2 opsin sequences from the European Eel were isolated, sequenced and expressed in vitro for an accurate measurement of their lambdamax values. In situ hybridisation revealed that glass eels express only rh2 opsin in their cone photoreceptors, while larger yellow eels continue to express rh2 opsin in the majority of their cones, but also have <5% of cones which express sws2 opsin. Silver eels showed the same expression pattern as the larger yellow eels. This observation was confirmed by qPCR (quantitative polymerase chain reaction). CONCLUSIONS: Larger yellow and silver European eels express two different cone opsins, rh2 and sws2. This work demonstrates that only the Rh2 cone opsin is present in younger fish (smaller yellow and glass), the sws2 opsin being expressed additionally only by older fish and only in <5% of cone cells. PMID- 20025775 TI - Developing obesity prevention interventions among minority ethnic children in schools and places of worship: The DEAL (DiEt and Active Living) study. AB - BACKGROUND: Childhood obesity is a major public health concern with serious implications for the sustainability of healthcare systems. Studies in the US and UK have shown that ethnicity is consistently associated with childhood obesity, with Black African origin girls in particular being more vulnerable to overweight and obesity than their White peers. Little is known, however, about what promotes or hinders engagement with prevention programmes among ethnic minority children. METHODS/DESIGN: This paper describes the background and design of an exploratory study conducted in London, UK. The aim of the study was to assess the feasibility, efficacy and cultural acceptability of child- and family-based interventions to reduce risk factors for childhood and adolescent obesity among ethnic minorities. It investigated the use of a population approach (in schools) and a targeted approach (in places of worship). We used a mixture of focus group discussions, in-depth interviews and structured questionnaires to explore what children, parents, grandparents, teachers and religious leaders think hinder and promote engagement with healthy eating and active living choices. We assessed the cultural appropriateness of validated measures of physical activity, dietary behaviour and self efficacy, and of potential elements of interventions informed by the data collected. We are also currently assessing the potential for wider community support (local councils, community networks, faith forums etc) of the intervention. DISCUSSION: Analysis of the data is ongoing but the emergent findings suggest that while the school setting may be better for the main implementation of healthy lifestyle interventions, places of worship provide valuable opportunities for family and culturally specific support for implementation. Tackling the rise in childhood and adolescent obesity is a policy priority, as reflected in a range of government initiatives. The study will enhance such policy by developing the evidence base about culturally acceptable interventions to reduce the risk of obesity in children. PMID- 20025776 TI - Hair mercury levels in Amazonian populations: spatial distribution and trends. AB - BACKGROUND: Mercury is present in the Amazonian aquatic environments from both natural and anthropogenic sources. As a consequence, many riverside populations are exposed to methylmercury, a highly toxic organic form of mercury, because of their intense fish consumption. Many studies have analysed this exposure from different approaches since the early nineties. This review aims to systematize the information in spatial distribution, comparing hair mercury levels by studied population and Amazonian river basin, looking for exposure trends. METHODS: The reviewed papers were selected from scientific databases and online libraries. We included studies with a direct measure of hair mercury concentrations in a sample size larger than 10 people, without considering the objectives, approach of the study or mercury speciation. The results are presented in tables and maps by river basin, displaying hair mercury levels and specifying the studied population and health impact, if any. RESULTS: The majority of the studies have been carried out in communities from the central Amazonian regions, particularly on the Tapajos River basin. The results seem quite variable; hair mercury means range from 1.1 to 34.2 microg/g. Most studies did not show any significant difference in hair mercury levels by gender or age. Overall, authors emphasized fish consumption frequency as the main risk factor of exposure. The most studied adverse health effect is by far the neurological performance, especially motricity. However, it is not possible to conclude on the relation between hair mercury levels and health impact in the Amazonian situation because of the relatively small number of studies. CONCLUSIONS: Hair mercury levels in the Amazonian regions seem to be very heterogenic, depending on several factors. There is no obvious spatial trend and there are many areas that have never been studied. Taking into account the low mercury levels currently handled as acceptable, the majority of the Amazonian populations can be considered exposed to methylmercury contamination. The situation for many of these traditional communities is very complex because of their high dependence on fish nutrients. It remains difficult to conclude on the Public Health implication of mercury exposure in this context. PMID- 20025777 TI - Complete reversal of epithelial to mesenchymal transition requires inhibition of both ZEB expression and the Rho pathway. AB - BACKGROUND: Epithelial to Mesenchymal Transition (EMT) induced by Transforming Growth Factor-beta (TGF-beta) is an important cellular event in organogenesis, cancer, and organ fibrosis. The process to reverse EMT is not well established. Our purpose is to define signaling pathways and transcription factors that maintain the TGF-beta-induced mesenchymal state. RESULTS: Inhibitors of five kinases implicated in EMT, TGF-beta Type I receptor kinase (TbetaRI), p38 mitogen activated protein kinase (p38 MAPK), MAP kinase kinase/extracellular signal regulated kinase activator kinase (MEK1), c-Jun NH-terminal kinase (JNK), and Rho kinase (ROCK), were evaluated for reversal of the mesenchymal state induced in renal tubular epithelial cells. Single agents did not fully reverse EMT as determined by cellular morphology and gene expression. However, exposure to the TbetaRI inhibitor SB431542, combined with the ROCK inhibitor Y27632, eliminated detectable actin stress fibers and mesenchymal gene expression while restoring epithelial E-cadherin and Kidney-specific cadherin (Ksp-cadherin) expression. A second combination, the TbetaRI inhibitor SB431542 together with the p38 MAPK inhibitor SB203580, was partially effective in reversing EMT. Furthermore, JNK inhibitor SP600125 inhibits the effectiveness of the TbetaRI inhibitor SB431542 to reverse EMT. To explore the molecular basis underlying EMT reversal, we also targeted the transcriptional repressors ZEB1 and ZEB2/SIP1. Decreasing ZEB1 and ZEB2 expression in mouse mammary gland cells with shRNAs was sufficient to up regulate expression of epithelial proteins such as E-cadherin and to re-establish epithelial features. However, complete restoration of cortical F-actin required incubation with the ROCK inhibitor Y27632 in combination with ZEB1/2 knockdown. CONCLUSIONS: We demonstrate that reversal of EMT requires re-establishing both epithelial transcription and structural components by sustained and independent signaling through TbetaRI and ROCK. These findings indicate that combination small molecule therapy targeting multiple kinases may be necessary to reverse disease conditions. PMID- 20025779 TI - Effect of filtration on morphine and particle content of injections prepared from slow-release oral morphine tablets. AB - BACKGROUND: Injections of mixtures prepared from crushed tablets contain insoluble particles which can cause embolisms and other complications. Although many particles can be removed by filtration, many injecting drug users do not filter due to availability, cost or performance of filters, and also due to concerns that some of the dose will be lost. METHODS: Injection solutions were prepared from slow-release morphine tablets (MS Contin) replicating methods used by injecting drug users. Contaminating particles were counted by microscopy and morphine content analysed by liquid chromatography before and after filtration. RESULTS: Unfiltered tablet extracts contained tens of millions of particles with a range in sizes from < 5 microm to > 400 microm. Cigarette filters removed most of the larger particles (> 50 microm) but the smaller particles remained. Commercial syringe filters (0.45 and 0.22 microm) produced a dramatic reduction in particles but tended to block unless used after a cigarette filter. Morphine was retained by all filters but could be recovered by following the filtration with one or two 1 ml washes. The combined use of a cigarette filter then 0.22 microm filter, with rinses, enabled recovery of 90% of the extracted morphine in a solution which was essentially free of tablet-derived particles. CONCLUSIONS: Apart from overdose and addiction itself, the harmful consequences of injecting morphine tablets come from the insoluble particles from the tablets and microbial contamination. These harmful components can be substantially reduced by passing the injection through a sterilizing (0.22 microm) filter. To prevent the filter from blocking, a preliminary coarse filter (such as a cigarette filter) should be used first. The filters retain some of the dose, but this can be recovered by following filtration with one or two rinses with 1 ml water. Although filtration can reduce the non-pharmacological harmful consequences of injecting tablets, this remains an unsafe practice due to skin and environmental contamination by particles and microorganisms, and the risks of blood-borne infections from sharing injecting equipment. PMID- 20025778 TI - Protocol for an experimental investigation of the roles of oxytocin and social support in neuroendocrine, cardiovascular, and subjective responses to stress across age and gender. AB - BACKGROUND: Substantial empirical evidence has demonstrated that individuals who are socially isolated or have few positive social connections seem to age at a faster rate and have more chronic diseases. Oxytocin is a neurohypophyseal hormone hypothesized to coordinate both the causes and effects of positive social interactions, and may be involved in positive physiological adaptations such as buffering the deleterious effects of stress and promoting resilience. The proposed research will examine whether and how oxytocin influences responses to stress in humans and will consider effects in relation to those of social support. METHODS/DESIGN: Experimental research will be used to determine whether exogenously administered oxytocin (intranasal) influences psychological and physiological outcomes under conditions of stress across gender and age in adulthood. Hypotheses to be tested are: 1) Oxytocin ameliorates the deleterious neuroendocrine, cardiovascular, and subjective effects of stress; 2) Oxytocin and social support have similar and additive stress-buffering effects; 3) Oxytocin effects are stronger in women versus men; and 4) Oxytocin effects are similar across a range of adult ages. Hypotheses will be tested with a placebo controlled, double-blind study using a sample of healthy men and women recruited from the community. Participants are randomly assigned to receive either oxytocin or placebo. They undergo a social stress manipulation with and without social support (randomly assigned), and outcome measures are obtained at multiple times during the procedure. DISCUSSION: Understanding the determinants of healthy aging is a major public health priority and identifying effective measures to prevent or delay the onset of chronic diseases is an important goal. Experimental research on oxytocin, social relationships, and health in adulthood will contribute to the scientific knowledge base for maximizing active life and health expectancy. At conclusion of the study we will have solid evidence concerning the effects of oxytocin on stress response and whether it has similar effects across age and gender groups. A neurobiological understanding of resilience can inform efforts for both prevention and intervention of diseases or problems common in later life. PMID- 20025780 TI - Correlation between expression of p53, p21/WAF1, and MDM2 proteins and their prognostic significance in primary hepatocellular carcinoma. AB - BACKGROUND: Tumor Protein p53 (p53), cyclin-dependent kinase inhibitor 1A (p21/WAF1), and murine double minute 2 (MDM2) participate in the regulation of cell growth. Altered expression of these gene products has been found in malignant tumors and has been associated with poor prognosis. Our aim was to investigate the expression of the 3 proteins in hepatocellular carcinoma (HCC) and their prognostic significance. METHODS: We examined p53, p21/WAF1, and MDM2 expression in 181 pairs of HCC tissues and the adjacent hepatic tissues by performing immunohistochemistry and examined the expression of the 3 proteins in 7 pairs of HCC tissues and the adjacent hepatic tissues by using western blot analysis. RESULTS: The expression of p53, p21/WAF1, and MDM2 in the HCC tissues was significantly higher than those in the adjacent hepatic tissues (P < 0.05). A statistical correlation was observed between p53 and p21/WAF1 expression in HCC tissues (R = 0.195, P = 0.008). A statistical correlation was observed between expression of p53 and p21/WAF1 (R = 0.380, P = 0.000), p53 and MDM2 (R = 0.299, P = 0.000), p21/WAF1 and MDM2 (R = 0.285, P = 0.000) in 181 liver tissues adjacent to the tumor. Patients with a low pathologic grade HCC (I+II) had a higher tendency to express p53 on tumor cells than the patients with high pathologic grade HCC (III+IV) (P = 0.007). Survival analysis showed that positive p21/WAF1 expression or/and negative MDM2 expression in HCC was a predictor of better survival of patients after tumor resection (P < 0.05). CONCLUSIONS: The proteins p53, p21/WAF1, and MDM2 were overexpressed in all the HCC cases in this study, and p53 and p21/WAF1 overexpression were positively correlated. The expression of p21/WAF1 and MDM2 can be considered as 2 useful indicators for predicting the prognosis of HCC. PMID- 20025781 TI - Nuclease-resistant double-stranded DNA controls or standards for hepatitis B virus nucleic acid amplification assays. AB - BACKGROUND: Identical blood samples tested using different kits can give markedly different hepatitis B virus (HBV) DNA levels, which can cause difficulty in the interpretation of viral load. A universal double-stranded DNA control or standard that can be used in all commercial HBV DNA nucleic acid amplification assay kits is urgently needed. By aligning all HBV genotypes (A-H), we found that the surface antigen gene and precore-core gene regions of HBV are the most conserved regions among the different HBV genotypes. We constructed a chimeric fragment by overlapping extension polymerase chain reaction and obtained a 1,349-bp HBVC+S fragment. We then packaged the fragment into lambda phages using a traditional lambda phage cloning procedure. RESULTS: The obtained armored DNA was resistant to DNase I digestion and was stable, noninfectious to humans, and could be easily extracted using commercial kits. More importantly, the armored DNA may be used with all HBV DNA nucleic acid amplification assay kits. CONCLUSIONS: The lambda phage packaging system can be used as an excellent expression platform for armored DNA. The obtained armored DNA possessed all characteristics of an excellent positive control or standard. In addition, this armored DNA is likely to be appropriate for all commercial HBV DNA nucleic acid amplification detection kits. Thus, the constructed armored DNA can probably be used as a universal positive control or standard in HBV DNA assays. PMID- 20025782 TI - A survey of attitudes toward clinical research among physicians at Kyoto University Hospital. AB - BACKGROUND: In Japan, only clinical research related to investigational new drug trials must be notified to regulatory bodies, and this lack of a uniform standard for clinical research has caused a number of difficulties. The objective of this study was to assess the willingness of physicians to participate in clinical research and to identify effective methods to promote and enhance clinical research. METHODS: We conducted a cross-sectional survey by administrating questionnaires to physicians in 31 departments in Kyoto University Hospital from October through November 2007. RESULTS: A total of 51.5% (310 of 602) of physicians completed the questionnaire. More than two-thirds of them reported currently participating in clinical research, and nearly all believed that clinical research is necessary for physicians. Less than 20% of respondents had specific training regarding clinical research, and most reported a need to acquire concepts and skills regarding clinical research, especially those related to statistics. "Paperwork was complicated and onerous" was the most frequently cited obstacle in conducting clinical research, followed by "few eligible patients" and "lack of time". Previous participation in and prospective participation in clinical research, previous writing a research protocol were positively associated with current participation in clinical research. CONCLUSIONS: Physicians in university hospitals need more training regarding clinical research, particularly in biostatistics. They also require administrative assistance. Our findings indicate that the quality of clinical research could be improved if training in clinical research methodology and biostatistics were provided, and if greater assistance in the preparation of study documents requested by the institutional Independent Ethics Committee were available. PMID- 20025783 TI - The International Research Society of Spinal Deformities (IRSSD) and its contribution to science. AB - From the time of its initial, informal meetings starting in 1980 to its formal creation in 1990, the IRSSD has met on a bi-annual basis to discuss all aspects of the spine and associated deformities. It has encouraged open discussion on all topics and, in particular, has tried to be the seed-bed for new ideas. The members are spread around the world and include people from all areas of academia as well as the most important people, the patients themselves. Most notably, application of the ideas and results of the research has always been at the forefront of the discussions. This paper was conceived with the idea of evaluating the impact made by the IRSSD over the last 30 years in the various areas and is intended to create discussion for the upcoming meeting in Montreal regarding future focus: "We are lost over the Atlantic Ocean but we are making good time." PMID- 20025785 TI - Prediction of backbone dihedral angles and protein secondary structure using support vector machines. AB - BACKGROUND: The prediction of the secondary structure of a protein is a critical step in the prediction of its tertiary structure and, potentially, its function. Moreover, the backbone dihedral angles, highly correlated with secondary structures, provide crucial information about the local three-dimensional structure. RESULTS: We predict independently both the secondary structure and the backbone dihedral angles and combine the results in a loop to enhance each prediction reciprocally. Support vector machines, a state-of-the-art supervised classification technique, achieve secondary structure predictive accuracy of 80% on a non-redundant set of 513 proteins, significantly higher than other methods on the same dataset. The dihedral angle space is divided into a number of regions using two unsupervised clustering techniques in order to predict the region in which a new residue belongs. The performance of our method is comparable to, and in some cases more accurate than, other multi-class dihedral prediction methods. CONCLUSIONS: We have created an accurate predictor of backbone dihedral angles and secondary structure. Our method, called DISSPred, is available online at http://comp.chem.nottingham.ac.uk/disspred/. PMID- 20025786 TI - [The effect of climate change on pollen allergy in the Netherlands]. AB - Climate change can exert a range of effects on pollen, which might have consequences for pollen-allergic patients. New allergenic pollen types might appear in the Netherlands, like common ragweed and olive, which result in allergy patients developing allergies that scarcely occur in the Netherlands at present. Trees, such as birches and planes, might produce larger quantities of pollen, which could result in more severe symptoms. The pollen season might become longer thereby extending the period in which patients suffer from allergy symptoms. This extension of the pollen season could be due to a prolonged flowering period of certain species, e.g. grasses, or the appearance of new species that flower in late summer, e.g. common ragweed. Climate change could cause an increase in heavy thunderstorms on summer days in the grass pollen season, which are known to increase the chance of asthma exacerbations. PMID- 20025784 TI - Characterizing the admixed African ancestry of African Americans. AB - BACKGROUND: Accurate, high-throughput genotyping allows the fine characterization of genetic ancestry. Here we applied recently developed statistical and computational techniques to the question of African ancestry in African Americans by using data on more than 450,000 single-nucleotide polymorphisms (SNPs) genotyped in 94 Africans of diverse geographic origins included in the HGDP, as well as 136 African Americans and 38 European Americans participating in the Atherosclerotic Disease Vascular Function and Genetic Epidemiology (ADVANCE) study. To focus on African ancestry, we reduced the data to include only those genotypes in each African American determined statistically to be African in origin. RESULTS: From cluster analysis, we found that all the African Americans are admixed in their African components of ancestry, with the majority contributions being from West and West-Central Africa, and only modest variation in these African-ancestry proportions among individuals. Furthermore, by principal components analysis, we found little evidence of genetic structure within the African component of ancestry in African Americans. CONCLUSIONS: These results are consistent with historic mating patterns among African Americans that are largely uncorrelated to African ancestral origins, and they cast doubt on the general utility of mtDNA or Y-chromosome markers alone to delineate the full African ancestry of African Americans. Our results also indicate that the genetic architecture of African Americans is distinct from that of Africans, and that the greatest source of potential genetic stratification bias in case-control studies of African Americans derives from the proportion of European ancestry. PMID- 20025787 TI - [Climate change and health]. AB - Despite the targets for greenhouse gas emissions agreed in Kyoto under the United Nations Framework Convention on Climate Change - again to be discussed in Copenhagen in December - climate change will still have serious effects on public health. The health effects of climate change will be noticeable also in the short run. Diseases which are transmitted by arthropod vectors will spread to more areas of the world than where they are present now. In addition, we will have to deal with allergies, deaths due to heat waves, diarrhoea and malnutrition. For this reason, every action is needed now in order to minimise the adverse effects on health. PMID- 20025788 TI - [Climate change and health in the Netherlands]. AB - Climate change possibly affects public health in the Netherlands, including changes in (a) temperature-related effects, heat stress and air pollution, (b) allergies, (c) vector borne infectious disease, and (d) food- and waterborne infectious disease. Due to many prevailing uncertainties, opinions differ regarding the exact size of the expected health risks and the speed at which these might occur, as well as regarding to what degree society would need to or could adapt to these potential health effects. Thus, the gaps in our knowledge are substantial. Scientists and experts are clearly concerned about the limited amount of attention being paid to health effects of climate change in the Netherlands. In response, a proposal for a research programme 'Klimaatverandering en Gezondheid' ('Climate change and health') has been developed over the past year. PMID- 20025789 TI - [Energy policy rather than climate policy]. AB - Energy policy and climate policy are two different issues and should not be treated as if they were the same. Whether the climate gets warmer or colder, saving energy and developing sustainable forms of energy production remain of paramount importance because fossil hydrocarbons are likely to be exhausted soon. But climate policy is a fallacy: it is human arrogance to think we can control the climate by reducing emissions and by storing CO2 underground. In spite of rising CO2 levels, the climate has cooled down slightly over the past decade. Since the International Panel on Climate Change (IPCC) did not predict this, it is questionable whether they can reliably predict warming. Other factors such as solar activity are probably more important for climate than greenhouse gases. The danger of coupling energy policy to climate policy is evident: if the climate cools down, people will lose belief in the greenhouse effect and therefore also lose interest in saving energy. PMID- 20025790 TI - [Dengue infection with fatal ending]. AB - A 54-year-old man was admitted with fever following a trip to St. Maarten in the Dutch Antilles, from where he had returned 6 days earlier. One year prior to this he had been treated on an Intensive Care Unit for a severe influenza A infection. Physical examination revealed remarkably cold extremities, furthermore there was metabolic acidosis, extreme haemoconcentration, prolongation of prothrombin time and activated partial thromboplastin time. His clinical condition deteriorated rapidly and he died the day after admission. Neither IgG nor IgM antibodies against dengue were found, but PCR on dengue virus RNA was positive. Over past decades there has been a strong increase in the number of dengue virus infections in travellers returning from Southeast Asia, Central and South America, the Caribbean and Africa. Whilst in general a primary dengue virus infection does not result in severe disease, this case illustrates the potentially fatal consequences of dengue in travellers. PMID- 20025791 TI - [Skin cancer epidemic in the Netherlands]. AB - Despite numerous warnings regarding the dangers of exposure to the sun, the number of skin cancer patients continues to increase rapidly. Dermatologists speak of a 'skin cancer epidemic' and estimate that the number of patients is substantially higher than the number estimated on the basis of cancer registry data. According to the Netherlands and Eindhoven cancer registries, 35.500 Dutch people were newly diagnosed with 'skin cancer' in 2006. Many skin cancer patients develop multiple skin tumours, therefore the total number of skin tumours is much higher than this. One in 6 Dutch people will develop skin cancer in their lifetime, 1 in 50 will develop a melanoma. Despite the good prognosis for most skin tumours, there is a high level of morbidity as a result of treatment. The large number of skin tumours and patients puts a great deal of pressure on the health care system. PMID- 20025792 TI - [Dengue: a growing risk to travellers to tropical and sub-tropical regions]. AB - Dengue is currently the most common arboviral infection worldwide. Due to global climate change and other factors, the vector of the virus - the Aedes mosquito - has spread considerably over the past decades. Dengue is endemic in almost all tropical and sub-tropical regions of the world; meaning that approximately 40% of the world population is at risk of acquiring a dengue infection. The clinical features of dengue vary from a non-specific febrile illness (dengue fever) to at times fatal serious conditions such as dengue haemorrhagic fever (DHF) and dengue shock syndrome (DSS). Considering the limited possibilities of prevention it is anticipated that the incidence of dengue will increase in the future. It is expected that health-care providers in non-endemic regions will encounter dengue infected patients with increasing frequency in their practices. PMID- 20025793 TI - [The climate debate: the facts]. AB - The first report by the Intergovernmental Panel on Climate Change (IPCC) appeared almost 20 years ago. Environmental contamination has a negative effect on the environment in which we live. However, the public at large is confused about the ins and outs of climate change. Managers, politicians, various kinds of advisors, scientists, so-called experts, sceptics and journalists have all taken it upon themselves to lead the debate. Whose task is it to ensure a sound discussion? Surely it is the IPCC's task. However, most politicians and many journalists, and even many scientists, do not take the trouble to read the entire IPCC report or parts of it. As a consequence, much nonsense is published and broadcast. An effective procedure to deal with the climate problem starts with a fair discussion of the scientific evidence. My advice is: just read the free IPCC report: http://www.ipcc.ch/ and click on 'WG I The Physical Science Basis'. PMID- 20025794 TI - Single-particle reconstruction of biological macromolecules in electron microscopy--30 years. AB - This essay gives the autho's personal account on the development of concepts underlying single-particle reconstruction, a technique in electron microscopy of macromolecular assemblies with a remarkable record of achievements as of late. The ribosome proved to be an ideal testing ground for the development of specimen preparation methods, cryo-EM techniques, and algorithms, with discoveries along the way as a rich reward. Increasingly, cryo-EM and single-particle reconstruction, in combination with classification techniques, is revealing dynamic information on functional molecular machines uninhibited by molecular contacts. PMID- 20025796 TI - Structure-functional intimacies of transient receptor potential channels. AB - Although a unifying characteristic common to all transient receptor potential (TRP) channel functions remains elusive, they could be described as tetramers formed by subunits with six transmembrane domains and containing cation-selective pores, which in several cases show high calcium permeability. TRP channels constitute a large superfamily of ion channels, and can be grouped into seven subfamilies based on their amino acid sequence homology: the canonical or classic TRPs, the vanilloid receptor TRPs, the melastatin or long TRPs, ankyrin (whose only member is the transmembrane protein 1 [TRPA1]), TRPN after the nonmechanoreceptor potential C (nonpC), and the more distant cousins, the polycystins and mucolipins. Because of their role as cellular sensors, polymodal activation and gating properties, many TRP channels are activated by a variety of different stimuli and function as signal integrators. Thus, how TRP channels function and how function relates to given structural determinants contained in the channel-forming protein has attracted the attention of biophysicists as well as molecular and cell biologists. The main purpose of this review is to summarize our present knowledge on the structure of channels of the TRP ion channel family. In the absence of crystal structure information for a complete TRP channel, we will describe important protein domains present in TRP channels, structure function mutagenesis studies, the few crystal structures available for some TRP channel modules, and the recent determination of some TRP channel structures using electron microscopy. PMID- 20025797 TI - From the Editors: dos and don'ts. PMID- 20025798 TI - Death is just not what it used to be. PMID- 20025795 TI - Elongation in translation as a dynamic interaction among the ribosome, tRNA, and elongation factors EF-G and EF-Tu. AB - The ribosome is a complex macromolecular machine that translates the message encoded in the messenger RNA and synthesizes polypeptides by linking the individual amino acids carried by the cognate transfer RNAs (tRNAs). The protein elongation cycle, during which the tRNAs traverse the ribosome in a coordinated manner along a path of more than 100 A, is facilitated by large-scale rearrangements of the ribosome. These rearrangements go hand in hand with conformational changes of tRNA as well as elongation factors EF-Tu and EF-G - GTPases that catalyze tRNA delivery and translocation, respectively. This review focuses on the structural data related to the dynamics of the ribosomal machinery, which are the basis, in conjunction with existing biochemical, kinetic, and fluorescence resonance energy transfer data, of our knowledge of the decoding and translocation steps of protein elongation. PMID- 20025799 TI - Not dead yet: controlled non-heart-beating organ donation, consent, and the Dead Donor Rule. PMID- 20025800 TI - Just caring: in defense of limited age-based healthcare rationing. PMID- 20025801 TI - Conscience-based exemptions for medical students. PMID- 20025802 TI - Actualizable potential, reproduction, and embryo research: bringing embryos into existence for different purposes or not at all. PMID- 20025803 TI - Consequentialism without consequences: ethics and embryo research. PMID- 20025804 TI - The ethics of aggressive discharge planning. PMID- 20025805 TI - Choosing deafness with preimplantation genetic diagnosis: an ethical way to carry on a cultural bloodline? PMID- 20025806 TI - The legal development of the informed consent doctrine: past and present. PMID- 20025807 TI - When is my genetic information your business? Biological, emotional, and financial claims to knowledge. PMID- 20025808 TI - Dignity: one, two, three, four, five, still counting. PMID- 20025809 TI - Clinical features and management of facial nerve paralysis in children: analysis of 24 cases. AB - OBJECTIVES: To evaluate the causes, treatment modalities and recovery rate of paediatric facial nerve paralysis. MATERIALS AND METHODS: We analysed 24 cases of paediatric facial nerve paralysis diagnosed in the otolaryngology department of Gachon University Gil Medical Center between January 2001 and June 2006. RESULTS: The most common cause was idiopathic palsy (16 cases, 66.7 per cent). The most common degree of facial nerve paralysis on first presentation was House-Brackmann grade IV (15 of 24 cases). All cases were treated with steroids. One of the 24 cases was also treated surgically with facial nerve decompression. Twenty-two cases (91.6 per cent) recovered to House-Brackmann grade I or II over the six month follow-up period. CONCLUSION: Facial nerve paralysis in children can generally be successfully treated with conservative measures. However, in cases associated with trauma, radiological investigation is required for further evaluation and treatment. PMID- 20025810 TI - Pharyngeal pouch surgery in north Glasgow: NICE (National Institute for Health and Clinical Excellence) practice or not? AB - AIM: To evaluate endoscopic pharyngeal pouch surgery practice in north Glasgow by comparing it to National Institute for Health and Clinical Excellence recommendations. METHODS: We reviewed the case notes of patients who had undergone pharyngeal pouch surgery from 1998 to 2008. Data obtained included patient demographics, procedures performed, complications and outcomes. RESULTS: One hundred patient case notes were reviewed. Surgical procedures performed included endoscopic stapling (n = 58), endoscopic laser surgery (26), external excision (one), cricopharyngeal myotomy (two) and pharyngoscopy with dilatation (three). Endoscopic stapling was abandoned in 10 patients (14.7 per cent), three of whom declined further surgery. There was a 2.2 per cent perforation rate for endoscopic procedures. Twenty-one per cent of patients required further surgery. CONCLUSION: Our practice was not in keeping with National Institute for Health and Clinical Excellence recommendations. Our complication rates were similar to other published series, although our rates for abandoned and revision procedures were higher. We suggest that pharyngeal pouch surgical procedures should be undertaken only by otolaryngologists with a primary head and neck interest. PMID- 20025811 TI - Cerebrospinal fluid otorrhoea: a rare presentation of Langerhans' cell histiocytosis of the temporal bone. AB - OBJECTIVE: To report a case of Langerhans cell histiocytosis of the temporal bone presenting with cerebrospinal fluid fistula. PATIENT: A Caucasian woman presented to a tertiary care centre in Quebec, Canada, with a new onset of cerebrospinal fluid fistula. She had a significant destructive lesion of the temporal bone, and was diagnosed with Langerhans cell histiocytosis on biopsy. INTERVENTIONS: The patient underwent surgical resection with reconstruction of the posterior fossa and tegmen. She suffered a relapse less than one year after surgery, and was finally treated with chemotherapy. MAIN OUTCOME AND RESULTS: The patient was free of disease at three-year follow up. No recurrence of the cerebrospinal fluid leak was observed after treatment. CONCLUSION: Langerhans cell histiocytosis of the temporal bone with intra-cranial involvement is rare in adults, with only two cases previously reported. Eleven paediatric cases have been reported. To our knowledge, this patient represents the first report of cerebrospinal fluid fistula as the initial presentation of the disease. PMID- 20025812 TI - Beware the silent presentation of a high and dehiscent jugular bulb in the external ear canal. AB - OBJECTIVE: We report two rare cases of high and dehiscent jugular bulbs protruding into the external ear canal; one case was silent, the other symptomatic. METHOD: The cases are presented and the relevant literature reviewed and discussed. RESULTS: A 52-year-old woman underwent mastoid exploration for suspected cholesteatoma. On raising the tympanomeatal flap, while still lateral to the annulus, brisk venous bleeding occurred from a high, dehiscent jugular bulb in the external ear canal. The second patient, a 42-year-old man, underwent imaging for suspected glomus tumour. A high, dehiscent jugular bulb was found protruding into the external ear canal and hypotympanum. CONCLUSION: High jugular bulbs in the middle ear have been reported many times. To our knowledge, there has only been one previous report of a high jugular bulb in the external ear. These cases illustrate the importance of full, accurate assessment of high resolution computed tomography images, and of appropriate, immediate management of bleeding. PMID- 20025813 TI - A low-cost, endoscopic, digital, still and video photography system for ENT clinics. AB - Image capture systems that display and record endoscopic images are important for documentation and teaching. We have modified a universal serial bus microscope to couple with most clinical endoscopes used in our practice. This very economical device produces images suitable for teaching, and potentially for clinical use. The implications of this could be significant for teaching, patient education, documentation and the developing world. PMID- 20025814 TI - Life-threatening carotid haemorrhage following blunt trauma. AB - INTRODUCTION: We report a case of internal carotid arterial damage following blunt neck trauma. This rare mechanism of injury demands a high index of suspicion to enable prompt specialist management. CASE REPORT: A 22-year-old man presented to hospital after sustaining blunt neck trauma. Rapid onset of stridor necessitated an emergency tracheostomy. Computed tomography angiography demonstrated a tear of the right internal carotid artery, which was repaired surgically. DISCUSSION: Blunt carotid vessel injury, although rare, has a high mortality rate. Mechanisms of injury include hyperextension and contralateral neck rotation, a direct blow to the vessel, and laceration by adjacent bony structures. The 'gold standard' investigation for suspected blunt carotid vessel injury is catheter angiography, although this carries a small risk of stroke. Computed tomography angiography is a less invasive, alternative investigation which has almost equivalent accuracy. The extent of damage to the vessel wall will dictate treatment. In our literature review, we discuss the presentation, investigation and different treatment modalities available. CONCLUSION: This case highlights an unusual mechanism of carotid artery injury, with a delayed, potentially fatal presentation. Such injury demands a high index of suspicion, and confirmation with specific investigations. Management is hazardous and requires experienced personnel in all aspects of care. PMID- 20025815 TI - Trends in paediatric airway surgery: a move towards endoscopic techniques. AB - The endoscope has long been an invaluable tool in assessment of the paediatric airway. Recently, its applications for definitive surgery of the airway have greatly increased, due to innovative surgical techniques, development of new instruments, improvements in anaesthesia and availability of new medications for endoscopic use. This review discusses the move towards endoscopic techniques in the management of paediatric airway disorders. PMID- 20025816 TI - Kainate receptors and signal integration by NG2 glial cells. AB - It is well established that NG2 cells throughout the young and adult brain consistently detect the release of single vesicles filled with glutamate from nearby axons. The released neurotransmitter glutamate electrically excites NG2 cells via non-NMDA (N-methyl-D-aspartic acid) glutamate receptors but the individual contribution of AMPA and kainate receptors to neuron-NG2 cell signalling, is not well understood. Here we pharmacologically block AMPA-type glutamate receptors and investigate whether hippocampal NG2 cells also express the kainate subtype of glutamate receptors and what may be their contribution to synaptic connectivity. It has been shown previously that vesicular glutamate release does not lead to a detectable activation of kainate receptors on NG2 cells. Here we report that while bath application of 250 nM-1 muM kainate does not have a major effect on NG2 cells it consistently induces a small and persistent depolarising current. This current was not mimicked by ATPA, suggesting that this current is carried by non-GluR5 containing kainate receptors. In addition to this inward current, nanomolar concentrations of kainate also produced a dramatic increase in the frequency of spontaneous GABA-A receptor-mediated synaptic currents (IPSCs) in NG2 cells. This increase in spontaneous IPSC frequency was even more pronounced on application of the GluR5 specific agonist ATPA (approximately 15-fold increase in frequency). In contrast, mono-synaptic stimulated IPSCs recorded in NG2 cells were unaffected by kainate receptor activation. Those and further experiments show that the occurrence of the high frequency of IPSCs is due to action potential firing of hippocampal interneurons caused by activation of GluR5 receptors on the somatodendritic membrane of the interneurons. Our data suggest that hippocampal kainate receptors are not only important for communication between neurons but may also play a dual and subtype-specific role for neuron-glia signalling: Firstly, extra-synaptic non GluR5 kainate receptors in the membrane of NG2 cells are ideally suited to instruct NG2 cells on the population activity of local excitatory neurons via ambient glutamate. Secondly, based on the known importance of GluR5 receptors on hippocampal interneurons for the generation of network rhythms and based on our finding that these interneurons heavily project onto NG2 cells, it appears that synaptic activation of interneuronal GluR5 receptors triggers signalling to NG2 cells which transmits the phase and frequency of ongoing network oscillations in the developing hippocampus. PMID- 20025817 TI - The role of glial cells in influencing neurite extension by dorsal root ganglion cells. AB - When pretreated with pertussis toxin (PTX), the neurites of adult rat dorsal root ganglion (DRG) cells in mixed cell cultures retract over a period of 2 h following the initial stimulus of removal from the cell culture incubator for brief periods of observation. The purpose of this investigation was to determine whether this PTX-dependent response was specific to any one of the three subpopulations of DRG neurons. However, no neurite retraction response was observed in neuron-enriched populations of cells, or in cultures enriched in isolectin B4 (IB4)-positive neurons or in IB4-negative neurons. But, the addition of non-neuronal cells, and/or medium conditioned by non-neuronal cells, was sufficient to restore the PTX-dependent neurite retraction response, but only in large diameter IB4-negative neurons. In conclusion, we have identified a regulatory response, mediated by Gi/o-proteins, which prevents retraction of neurites in large diameter IB4-negative cells of adult rat DRG. The non-neuronal cells of adult rat DRG constitutively release factor/s that can stimulate neurite retraction of a subset of isolated DRG neurons, but this property of non-neuronal cells is only observed when the Gi/o-proteins of large diameter IB4-negative cells are inhibited. PMID- 20025818 TI - Abiotic versus biotic hierarchies in the assembly of parasite populations. AB - The presence or absence of parasites within host populations is the result of a complex of factors, both biotic and abiotic. This study uses a non-parametric classification tree approach to evaluate the relative importance of key abiotic and biotic drivers controlling the presence/absence of parasites with complex life cycles in a sentinel, the common killifish Fundulus heteroclitus. Parasite communities were classified from 480 individuals representing 15 fish from 4 distinct marsh sites in each of 4 consecutive seasons between 2006 and 2007. Abiotic parameters were recorded at continuous water monitoring stations located at each of the 4 sites. Classification trees identified the presence of benthic invertebrate species (Gammarus sp. and Littorina sp.) as the most important variables in determining parasite presence: secondary splitters were dominated by abiotic variables including conductance, pH and temperature. Seventy percent of hosts were successfully classified into the correct category (infected/uninfected) based on only these criteria. The presence of competent definitive hosts was not considered to be an important explanatory variable. These data suggest that the most important determinant of the presence of these parasite populations in the common killifish is the availability of diverse communities of benthic invertebrates. PMID- 20025819 TI - Inhibition of cytochrome P450-mediated metabolism enhances ex vivo susceptibility of Fasciola hepatica to triclabendazole. AB - A study has been carried out to investigate whether the action of triclabendazole (TCBZ) against Fasciola hepatica is altered by inhibition of drug metabolism. The cytochrome P450 (CYP P450) system was inhibited using piperonyl butoxide (PB). The Oberon TCBZ-resistant and Cullompton TCBZ-susceptible isolates were used for these experiments. The CYP P450 system was inhibited by a 2 h pre-incubation in PB (100 mum). Flukes were then incubated for a further 22 h in NCTC medium containing either PB; PB+nicotinamide adenine dinucleotide phosphate (NADPH) (1 nm); PB+NADPH+TCBZ (15 microg/ml); or PB+NADPH+TCBZ.SO (15 microg/ml). Morphological changes resulting from drug treatment and following metabolic inhibition were assessed using scanning electron microscopy. After treatment with either TCBZ or TCBZ.SO alone, there was greater disruption to the TCBZ susceptible than the resistant isolate. However, co-incubation with PB and TCBZ/TCBZ.SO lead to more severe surface changes to the TCBZ-resistant Oberon isolate than with each drug on its own. With the TCBZ-susceptible Cullompton isolate, there was limited potentiation of drug action, and only with TCBZ.SO. The results support the concept of altered drug metabolism in TCBZ-resistant flukes and this process may play a role in the development of drug resistance. PMID- 20025820 TI - Four new species of KudoaMeglitsch, 1947 (Myxosporea: Multivalvulida) from Australia with recommendations for species descriptions in the Kudoidae. AB - Kudoid parasites are significant pathogens of marine fish. In the past, specific identification has been difficult due to a paucity of detailed morphological and biological information provided in the original description of some species. However, the introduction of DNA analysis has dramatically improved diagnosis. For morphological characterization, this paper proposes a new set of schematics including additional measurements to give a more thorough description of spore morphology, and provides evidence for uniqueness of 4 new species: Kudoa gunterae n. sp. (from 10 pomacentrid species and 1 apogonid species), K. kenti n. sp. (from 4 pomacentrid species), K. paraquadricornis n. sp. (from 4 carangid species), and K. whippsi n. sp. (from 8 pomacentrid species and 1 apogonid species). Subtle morphological differences found between closely related species were reflected in their genetics, with increased resolution provided by the large subunit, compared with that of the small subunit, of the ribosomal DNA gene region. This article proposes comprehensive requirements for species descriptions within the Kudoidae that incorporate biology, morphology, and genetic sequence. PMID- 20025821 TI - The influence of human settlements on the parasite community in two species of Peruvian tamarin. AB - Although there is a growing recognition that the transfer of diseases between humans and non-human primates can be of great significance for conservation biology, there have been only a few studies focusing on parasites. In this study, saddleback (Saguinus fuscicollis) and moustached tamarin (Saguinus mystax) from the rainforest of the Peruvian lowlands were used as models to determine helminth parasite associations between canopy-dwelling primate species and a nearby human settlement. The human population showed high prevalences of infestation with a number of nematodes, including Ascaris lumbricoides (88.9%), Trichuris trichiura (37%) and hookworms (55.6%). However, the ova of these geohelminths were not detectable in tamarin faeces. Thus, no direct parasite transfer from humans to non-human primates could be documented. However, tamarin groups with more frequent contact to humans and their facilities had significantly higher prevalences and egg output of Prosthenorchis elegans, an important primate pathogen, than a forest group. In contrast, a cestode was significantly more common with more egg output in sylvatic than in human-associated groups. Human alteration of the habitat is likely to play a major role in determining the occurrence, prevalence and intensity of helminth infestation of wild non-human primates. PMID- 20025822 TI - Contrasting detachment strategies in two congeneric ticks (Ixodidae) parasitizing the same songbird. AB - In non-permanent parasites the separation from the host should take place in suitable habitats that allow the continuation of their life cycle. Furthermore, detachment strategies determine the parasites' dispersal capability, a characteristic on which epidemiological dynamics and the evolution of host specificity centre. In this study we experimentally investigate in the laboratory how 2 congeneric tick species, with contrasting habitat requirements, time detachment from one of their current songbird hosts (Parus major). Ixodes arboricola is a nidicolous tick, infesting bats and birds breeding or roosting in tree holes. Ixodes ricinus is a non-nidicolous generalist that parasitizes mammals, birds and even reptiles. We experimentally infested full-grown great tits, P. major, and found that I. arboricola detaches during the night, the moment when P. major sleeps in tree holes. In contrast, I. ricinus detaches during the day, the moment when birds are most active. In addition we found that all I. ricinus immatures left the birds within 5.5 days, while in I. arboricola the detachment time was long (up to 20 days) and highly variable. We discuss these findings with respect to their implications on the ticks' dispersal capability and host specificity. PMID- 20025823 TI - Inhibitory effects of (-)-epigallocatechin-3-gallate from green tea on the growth of Babesia parasites. AB - (-)-Epigallocatechin-3-gallate (EGCG) is the major tea catechin and accounts for 50-80% of the total catechin in green tea. (-)-Epigallocatechin-3-gallate has antioxidant, anti-inflammatory, anti-microbial, anti-cancer, and anti trypanocidal activities. This report describes the inhibitory effect of (-) Epigallocatechin-3-gallate on the in vitro growth of bovine Babesia parasites and the in vivo growth of the mouse-adapted rodent babesia B. microti. The in vitro growth of the Babesia species was significantly (P<0.05) inhibited in the presence of micromolar concentrations of EGCG (IC50 values=18 and 25 microM for B. bovis, and B. bigemina, respectively). The parasites showed no re-growth at 25 microM for B. bovis and B. bigemina in the subsequent viability test. The drug significantly (P<0.05) inhibited the growth of B. microti at doses of 5 and 10 mg/kg body weight, and the parasites completely cleared on day 14 and 16 post inoculation in the 5 and 10 mg/kg treated groups, respectively. These findings highlight the potentiality of (-)-Epigallocatechin-3-gallate as a chemotherapeutic drug for the treatment of babesiosis. PMID- 20025824 TI - The genomic Echinococcus microsatellite EmsB sequences: from a molecular marker to the epidemiological tool. AB - In the field of molecular and epidemiological parasitology, characterization of fast evolving genetic markers appears as an important challenge to consider the diversity and genetic structure of parasites. The study of respective populations can help us to understand their adaptive strategies to survive and perpetuate the species within different host populations, all trying to resist infection. In the past, the relative monomorphic features of Echinococcus multilocularis, the causative agent of alveolar echinococcosis and a severe human parasitic disease, did not stimulate studies dealing with the genetic variability of Echinococcus species or respective populations. A recently developed, characterized and validated original multilocus microsatellite, named EmsB, tandemly repeated in the genome, offered an additional opportunity for this line of investigation. We have compiled in this review new insights brought by this molecular tracker on the transmission activity of Echinococcus among different hosts and at different geographical scales. PMID- 20025825 TI - The selection of experimental doses and their importance for parasite success in metacercarial infection studies. AB - Experimental studies of parasite transmission are essential for advances in basic and applied parasitology. A survey of the results of published experiments can identify the determinants of both variation among studies in experimental design and of parasite infection success. Here, analyses are conducted on data compiled from a total of 106 metacercarial infection experiments (35 on Echinostomatidae, 37 on Fasciolidae, 34 on other trematodes) obtained from 83 studies. All of these involved experimental oral infection of individual definitive hosts by a single known dose of metacercariae under controlled conditions. Across these studies, the metacercarial dose used (i) was typically about 10 times higher than the average natural dose that could be acquired by feeding on intermediate hosts (for taxa other than Fasciolidae), and (ii) showed a positive relationship with the body mass of the definitive host, although this relationship was only significant for Fasciolidae. Although the chosen dose was rarely justified, the larger the definitive host, the more metacercariae it received. Among Echinostomatidae and Fasciolidae, there was also a significant dose-dependent effect on infection success: the higher the dose used in an experiment, the smaller the proportion of metacercariae recovered from the host. This effect was mitigated by definitive host body mass, with infection success being generally lower in larger definitive hosts. For Echinostomatidae, the taxonomic identity of the definitive host also mattered, with metacercariae achieving higher infection success in mammals than in birds. The present findings suggest that the design of experimental infection studies requires greater consideration if their results are to yield useful biological insights. PMID- 20025826 TI - The potential of mass spectrometry for the global profiling of parasite metabolomes. AB - The strengths and limitations of existing mass spectrometry methods for metabolite detection and identification are discussed. A brief review is made of the methods available for quenching and extraction of cells or organisms prior to instrumental analysis. The techniques available for carrying out mass spectrometry-based profiling of metabolomes are discussed using the analysis of extracts from trypanosomes to illustrate various points regarding methods of separation and mass spectrometric analysis. The advantages of hydrophilic interaction chromatography (HILIC) for the analysis of polar metabolites are discussed. The challenges of data processing are outlined and illustrated using the example of ThermoFisher's Sieve software. The existing literature on applications of mass spectrometry to the profiling of parasite metabolomes is reviewed. PMID- 20025827 TI - Induction and regulation of Trypanosoma brucei VSG-specific antibody responses. AB - The review addresses how infection with Trypanosoma brucei affects the development, survival and functions of B lymphocytes in mice. It discusses (1) the contributions of antibodies to trypanosome clearance from the bloodstream, (2) how B lymphocytes, the precursors of antibody producing plasma cells, interact with membrane form variable surface glycoprotein (VSG), i.e. with monovalent antigen that is free to diffuse within the lipid bilayer of the trypanosome plasma membrane and consequently can cross-link B cell antigen specific receptors by indirect processes only and (3) the extent and underlying causes of dysregulation of humoral immune responses in infected mice, focusing on the impact of wild type and GPI-PLC-/- trypanosomes on bone marrow and extramedullary B lymphopoiesis, B cell maturation and survival. PMID- 20025828 TI - Immunity-mediated regulation of fecundity in the nematode Heligmosomoides polygyrus--the potential role of mast cells. AB - Previous studies have shown that host immunity regulates the fecundity of nematodes. The present study was aimed at clarifying the reversible nature of fecundity in response to changes of immunological status and to determine which effector cells are responsible for compromising fecundity in Heligmosomoides polygyrus. Enhanced fecundity was observed in immunocompromised SCID and nu/nu mice compared to those in the corresponding wild-type mice, with significantly fewer numbers of intrauterine eggs produced in the wild-type than in the immunodeficient mice. When 14-day-old adult worms from BALB/c mice were transplanted into naive BALB/c mice, their fecundity increased significantly as early as 24 h post-transplantation, but not when they were transferred into immune mice, suggesting the plastic and reversible nature of fecundity in response to changes in host immunological status. In mast cell-deficient W/W(v) mice, nematode fecundity was significantly higher than in mast cell-reconstituted W/W(v) or +/+ mice. The serum levels of the mast-cell protease mMCP1 were markedly increased in the wild-type as well as the mast cell-reconstituted W/W(v), but not in the W/W(v), SCID, or nu/nu mice during infection. These findings raise the interesting possibility that certain activities of mast cells, either directly or indirectly, regulate parasite fecundity during infection. PMID- 20025829 TI - Lifestyle risk factors for overweight in Japanese male college students. AB - OBJECTIVE: To identify lifestyle and sociodemographic risk factors of overweight among male college students. DESIGN: A retrospective cohort study. SETTING: Annual health checkup in a single university in Japan. PARTICIPANTS: Male students who underwent two successive health checkups from their third school year between 2000 and 2007 and whose BMI at baseline of this study was 22.0 kg/m2 or more (n 4634). RESULTS: During the 1-year follow-up, 598 students (12.9 %) reached the study endpoint, i.e. more than a 5 % increase in BMI. Independent risk factors for substantial BMI increase included infrequent exercise (OR = 1.33; 95 % CI 1.11, 1.60), no or infrequent alcohol drinking (OR = 1.30; 95 % CI 1.08, 1.57), frequently skipping breakfast (OR = 1.34; 95 % CI 1.12, 1.61), preference for fatty food (OR 1.36; 95 % CI 1.04, 1.78) and living alone (OR = 1.23; 95 % CI 0.99, 1.52). Students were readily stratified according to risk for substantial BMI gain by counting the number of their risk factors. OR (95 % CI) for the risk between the no risk factor group and students with two, three, four and five risk factors were 1.61 (0.96, 2.70), 2.24 (1.34, 3.75), 2.42 (1.39, 4.23) and 6.22 (2.58, 15.0), respectively. CONCLUSION: These data suggest that avoidance of certain risk factors in college life is associated with a decrease in incidence of overweight among male students. PMID- 20025830 TI - Peanut consumption and cardiovascular risk. AB - OBJECTIVE: We evaluated the effects of peanut consumption on lipid profiles, atherogenic index of plasma (AIP) and CHD risk in hypercholesterolaemic men. DESIGN: Randomised crossover clinical trial. SETTING: Participants were randomly assigned to two groups. They were asked to consume peanut supplements (about 77 g) with their habitual diet for 4 weeks. SUBJECTS: Fifty-four hypercholesterolaemic men with total cholesterol (TC) concentrations between 200 and 350 mg/dl. RESULTS: Compared with the habitual diet, peanut supplementation of the habitual diet significantly reduced TC/HDL cholesterol (HDL-C) ratio (mean 1 (se 0.3) P = 0.001) and LDL cholesterol (LDL-C)/HDL-C ratio (mean 0.7 (se 0.2); P = 0.001). Peanut consumption increased HDL-C (mean 6.1 (se 1.5) mg/dl; P < 0.001) and total antioxidant capacity (TAC) (mean 1.2 (se 0.6) U/mL P = 0.04). In addition, peanut consumption significantly reduced the AIP (mean 0.1 (se 0.03) P = 0.01) and CHD estimated risk over 10 years based on systolic and diastolic blood pressures (mean 1.4 % (se 0.5 %) P = 0.004 and mean 2.2 % (se 0.5 %) P < 0.001, respectively). CONCLUSIONS: Short-term peanut consumption might improve lipid profiles, the AIP and CHD risk in free-living hypercholesterolaemic men. PMID- 20025831 TI - Yoghurt and dairy snacks presented for sale to an Australian consumer: are they becoming less healthy? AB - OBJECTIVE: To assess the nutrient profile of yoghurts and dairy desserts. DESIGN: Nutrition information panels and product labels on yoghurts and dairy desserts offered for sale were surveyed in 2005 and 2008 and nutrients analysed by two nutrient profiling systems. SETTING: A large supermarket in metropolitan Melbourne, Australia. RESULTS: In total, 248 and 140 dairy snacks (yoghurt, fromage frais or dairy desserts) were surveyed in 2005 and 2008, respectively. Over this time, median packet size rose significantly (P < or = 0.001). In yoghurts, median energy and total fat content also increased while protein decreased (all P < 0.05). The proportion of 'full-fat' products rose from 36 % to 46%. Because of the addition of sugar, most 'reduced-fat' yoghurts had energy content similar to many 'full-fat' yoghurts. Overall, the proportion of yoghurts and dairy desserts that were 'less healthy' (i.e. displaying one or more 'red traffic lights' for high fat, saturated fat, salt and sugar content) rose from 12% in 2005 to 23% in 2008. Only 1-2% could be deemed 'healthy' by the most stringent criterion (displaying four 'green traffic lights'), while 21% (2005) or 28% (2008) were 'healthy' by a nutrient profiling system that included a score for protein. Sucrose, the most common sweetener, was found in levels up to 29 g/100 g. Claims on packaging mainly related to Ca, fat or protein content. Few labels referred to sugar content. CONCLUSIONS: The deterioration in nutrient quality of yoghurts needs to be redressed. PMID- 20025832 TI - Non-dieting group interventions for overweight and obese women: what predicts non completion and does completion improve outcomes? AB - OBJECTIVE: To determine factors which predict non-completion of group non-dieting interventions for overweight women, and to investigate whether completion improves outcomes. DESIGN: First, baseline predictors of non-completion were identified; then changes at 10 weeks and 12 months were compared between completers and non-completers of 10-week non-dieting interventions. SETTING: General community. SUBJECTS: Participants were 119 women (aged 25-65 years, BMI > or = 28 kg/m2) with at least one cardiovascular risk factor. Participants who attended at least eight of the ten sessions were classified as completers, and non-completers were those who attended fewer than eight sessions. Measures included BMI, blood pressure, psychological distress, lifestyle behaviours and eating self-efficacy. RESULTS: Logistic regression analyses indicated that women were less likely to be non-completers at non-dieting group programmes if, at baseline, they were more highly educated or had healthier nutrition behaviours (controlling for education). Only healthier nutrition behaviour was negatively associated with non-completion in the final model. Twelve months after the intervention, completers showed significantly greater improvements in body weight (mean change -0.53 kg), systolic and diastolic blood pressure (-6.3 and -4.1 mmHg, respectively), stress management behaviour score (+0.5) and psychotic symptoms score (-0.1) than non-completers (all P < 0.05). CONCLUSIONS: Highly educated women already engaging in some healthier lifestyle choices were less likely to be non-completers in non-dieting group programmes. Since important treatment outcomes vary according to attendance, future trials of non-dieting interventions should report the effects of completion on outcomes. PMID- 20025833 TI - 6-n-Propylthiouracil sensitivity and obesity status among ethnically diverse children. AB - OBJECTIVE: To examine the relationship of 6-n-propylthiouracil (PROP) sensitivity to BMI while statistically controlling for demographic characteristics in two age groups of children: 9-10 years and 17-18 years (n 1551). DESIGN: Cross-sectional design with a multi-ethnic (White, African-American, Hispanic, Other) sample of 813 children aged 9-10 years and 738 children aged 17-18 years. Children were recruited from local elementary and high schools with at least 30 % minority ethnic enrolment. Children's height, weight and waist circumference were measured along with their PROP taster status. PROP was measured using two paper discs, one impregnated with NaCl (1.0 mol/l) and the other with PROP solution (0.50 mmol/l). RESULTS: A significant PROP sensitivity by socio-economic status (SES) interaction term (P = 0.010) was detected wherein supertasters had the largest BMI percentile and Z-score, but only among the group with highest SES. CONCLUSIONS: The results suggest that other factors overwhelmed the influence of PROP sensitivity on adiposity in lower-SES groups. The percentage of variance accounted for by the interaction term was about 1 %. Thus, PROP supertasters had the largest BMI percentile and Z-score, but only among the highest-SES group. PMID- 20025834 TI - Optimising waiting: a view from the English National Health Service. AB - Recent authors have proposed that waiting times for elective treatment should be reduced to the point where the costs of doing so exceed the benefits. This paper considers how this criterion could be put into effect. Taking benefits first it argues that these could be estimated in three different ways - social cost benefit, clinical and user valuation - that would not necessarily produce consistent results and hence a choice has to be made between them. It then considers the costs of reducing waits and argues, citing relevant evidence, that these may range widely according to whether or not reductions can be achieved through simple management measures or whether more long-term capacity is required. It concludes therefore that the apparently simple criterion proposed for defining the point where waiting times are optimal is hard to establish. Choice of criterion must be made in the light of the overall values that a given health care system is intended to promote. PMID- 20025835 TI - Dried leaf extract of Olea europaea ameliorates islet-directed autoimmunity in mice. AB - The health-promoting effects of various constituents of the olive tree (Olea europaea) are mainly associated with hypoglycaemic and insulin-sensitising activities and have been widely demonstrated in the metabolic syndrome and type 2 diabetes. However, their biological activity in autoimmune type 1 diabetes (T1D) is poorly characterised. Therefore, the influence of O. europaea-derived components present in dry olive leaf extract (DOLE) was examined in two established preclinical models of human T1D, which differ in some aspects of diabetogenesis: multiple low-dose streptozotocin-induced diabetes in susceptible C57BL/6 and CBA/H mouse strains; cyclophosphamide-accelerated diabetes in non obese diabetic mice. In both T1D models, in vivo administration of DOLE significantly reduced clinical signs of diabetes (hyperglycaemia and body weight loss) and led to complete suppression of histopathological changes in pancreatic islets. In line with these, insulin expression and release were restored in DOLE treated mice. Interestingly, inducible NO synthase expression and NO production were significantly elevated in peripheral tissues but were down-regulated within the local environment of the endocrine pancreas. This interference was reflected in NO-mediated suppression of T lymphocyte proliferation and lower production of the proinflammatory cytokines interferon-gamma, IL-17 and TNF-alpha in the spleen, with subsequent blockade of beta-cell destruction. The results suggest that DOLE interferes with development of autoimmune diabetes by down-regulating production of proinflammatory and cytotoxic mediators. Therefore, the potential use of a DOLE-enriched diet for prophylaxis/treatment of human T1D, and possibly other autoimmune diseases, is worthy of further investigation. PMID- 20025836 TI - Group versus individual cognitive treatment for obsessive-compulsive disorder: Changes in severity at post-treatment and one-year follow-up. AB - BACKGROUND: Very few studies have compared the efficacy of individual and group cognitive behaviour therapy (CBT) for the treatment of Obsessive-Compulsive Disorder (OCD) by taking into consideration the change in OCD severity in both the short and long term. AIMS: To conduct an open trial of individual versus group CBT for OCD, comparing the clinical and statistically significant changes in severity both at post-treatment and one year later. METHOD: Forty-two OCD subjects were assigned to individual (n = 18) or group CBT (n = 24, in four groups). Sixteen and 22 subjects completed the treatment in the individual and group conditions, respectively. The Yale-Brown Obsessive Compulsive Scale was recorded at pre-treatment, post-treatment and at the one-year follow-up. RESULTS: At the end of treatment, the clinically significant change was comparable for the two treatment conditions and remained stable at the one-year follow-up. Of the 16 participants who completed the individual CBT treatment, 68.75% were classified as recovered at post-treatment, compared to 40.9% of those receiving group CBT. At follow-up the rate of recovery decreased to 62.5% in individual CBT and to 31.8% in group CBT. CONCLUSIONS: Group CBT is effective in decreasing OCD severity. The post-treatment changes were maintained one year later. Nevertheless, these changes were higher in the individual delivery of CBT. PMID- 20025837 TI - Multicentre randomised controlled trial examining the cost-effectiveness of contrast-enhanced high field magnetic resonance imaging in women with primary breast cancer scheduled for wide local excision (COMICE). AB - OBJECTIVES: To determine whether the addition of magnetic resonance imaging (MRI) to current patient evaluation by triple assessment would aid tumour localisation within the breast and thus reduce the reoperation rate in women with primary breast tumours who are scheduled for wide local excision (WLE), and to assess whether the addition of MRI would be cost-effective for the UK NHS. DESIGN: A multicentre, randomised controlled, open, parallel group trial with equal randomisation. The main design was supplemented with a qualitative study to assess patients' experiences of the treatment process and care pathway, and involved the development of a non-scheduled standardised interview (NSSI). SETTING: The study took place at 45 hospitals throughout the UK. PARTICIPANTS: Women aged 18 years or over with biopsy-proven primary breast cancer who had undergone triple assessment, were scheduled for WLE, and were capable of providing written informed consent. INTERVENTIONS: Patients were randomised to receive MRI or no MR1. Randomisation was performed using minimisation, incorporating a random element. All MRI was performed at 1.5 T or 1.0 T with a dedicated bilateral breast coil. MAIN OUTCOME MEASURES: The primary end point of the trial was the reoperation rate. Secondary outcome measures included discrepancies between imaging and histopathology, and the effectiveness of using both procedures; change in clinical management after using MRI; the clinical significance of MRI-only-detected lesions; the rate of interventions; the ipsilateral tumour recurrence rate; patient quality of life (QoL); and cost effectiveness. RESULTS: From a total of 1623 patients, 816 were randomised to MRI and 807 to no MRI. No differences in reoperation rates were found between the two groups of patients [MRI patients 18.75%, no MRI 19.33%, difference 0.58%, 95% confidence interval (CI) -3.24 to 4.40]. Therefore, the addition of MRI to conventional triple assessment was not found to be statistically significantly associated with a reduced reoperation rate (odds ratio = 0.96, 95% CI 0.75-1.24, p = 0.7691). The best agreement between all imaging modalities and histopathology with regard to tumour size and extent of disease was found in patients over 50 years old with ductal tumours NST and who were node negative. In the imaging arm, mastectomy was found to be pathologically avoidable for 16 (27.6%) out of 58 patients who underwent the procedure. There were no significant differences between the groups regarding the proportion of patients receiving chemotherapy, radiotherapy or additional adjuvant therapies, as well as for local recurrence free interval rates and QoL. An acceptable NSSI was developed for use in this population of patients. Economic analysis found no difference in outcomes between the two trial arms. CONCLUSIONS: The addition of MRI to triple assessment did not result in a reduction in operation rates, and the use of MRI would thus consume extra resource with few or no benefits in terms of cost-effectiveness or HRQoL. However, MRI showed potential to improve tumour localisation, and preoperative biopsy of MRI-only-detected lesions is likely to minimise the incidence of inappropriate mastectomy. TRIAL REGISTRATION: Current Controlled Trials ISRCTN57474502. PMID- 20025838 TI - Second meeting of the French CEIP (Centres d'Evaluation et d'Information sur la Pharmacodependance). Part I: how to evaluate and prevent the abuse and dependence on hypnotic/anxiolytic drugs? AB - The second meeting of the French CEIP (Centres d'Evaluation et d'Information sur la Pharmacodependance) was organized during the annual congress of the French Society of Pharmacology Therapeutics and Physiology in 2008. The aim of this meeting was to update the knowledge on abuse and dependence of the anxiolytics and hypnotics from different points of view (pharmacoepidemiology, epidemiology and treatment). The first part of this meeting summarized the pharmacological data obtained by the pharmacoepidemiological tools developed by the CEIP network. Even if the abuse liability of these agents is not a new problem, it remains always present and characterized by differences of misuse between drugs in real life settings. The second part targeted to a subtype of consumers, the elderly population, because older patients are more likely to be prescribed with multiple prescriptions and also more at risk for prescription abuse. Despite this evidence, there is a scarcity of information on the factors associated with a such behaviour and its screening, assessment, diagnosis and treatment. PMID- 20025839 TI - Second meeting of the French CEIP (Centres d'Evaluation et d'Information sur la Pharmacodependance). Part II: benzodiazepine withdrawal. AB - The aim of this meeting was to describe the news trends about abuse and addiction of anxiolytics and hypnotics. The part II of this meeting reviewed several aspects of the withdrawing benzodiazepines focusing in particular on clinical symptoms, biological patterns, and strategies for discontinuation. The discontinuation is usually beneficial due to the importance of adverse effects induced by long-term use of benzodiazepines. Several clinicals options have been described including gradual tapering of the current benzodiazepine, substitution with a long acting benzodiazepines or treating the symptoms of withdrawal. Psychological interventions range from a simple support through counselling to expert cognitive-behavioural therapy. However more controlled clinical trials are needed to promote adequate and optimal patient care in management of benzodiazepine withdrawing. PMID- 20025840 TI - [Influential factors in regular and intensive use of psychoactive drugs]. AB - The article attempts to construct a typology to the use of psychoactive drugs and the study of the various factors linked to their use, whether heavy or regular. The results draw on the exploitation of databases containing medicines presented for reimbursement by medical insurance in the Nord - Pas-de-Calais region. In 2007 and 2008, 20.7% of the insured population (764,650 people) benefited from reimbursements for this type of medication. Among the beneficiaries, about a third (30.5%) had an intense consummation. Age, the CMUs benefits and the fact that a person is or not followed by a psychiatrist are the most influential variables on the intensity and the regularity of intake. The observed regularity of the recourse to benzodiazepines does not conform with the current recommendations. PMID- 20025841 TI - Therapeutic influence of zinc and ascorbic acid against lead induced biochemical alterations. AB - The influence of co-administration of zinc (10 mg/kg, intraperitoneal [ip]) and ascorbic acid (10, 20 and 30 mg/kg, ip) against lead (lead acetate; 35 mg/kg, ip for 3 days) induced biochemical alterations was studied in young albino rats. The results revealed significant fall in hemoglobin content, on the other hand significant raise in the activity of serum transaminases and serum alkaline phosphatase after lead administration. Significant increase in lipid peroxidation and decreased level of reduced glutathione in liver showed oxidative stress due to lead exposure. Total protein content in liver and kidney were diminished after lead exposure. Activity of acid phosphatase in liver and kidney and alkaline phosphatase in kidney was increased significantly. Zinc and ascorbic acid treatment showed moderate therapeutic efficacy when administered individually, whereas more pronounced protective effects were observed after combined therapy of zinc and different doses of ascorbic acid. The results thus, suggested that co administration of zinc and ascorbic acid may be useful in restoration of lead induced biochemical alterations. PMID- 20025842 TI - PUVA therapy and narrowband UVB therapy in Tunisian patients with mycosis fungoides. AB - BACKGROUND: PUVA therapy and narrowband UVB therapy (NBUVB) constitute a widely used first line therapy in mycosis fungoides (MF). METHODS: It is a retrospective study dealing with 7 patients with MF treated by phototherapy [3 with PUVA (III, IV, V phototype) and 4 with NBUVB (III-IV phototype)]. One patient had both treatments (PUVA then NBUVB for a relapse). RESULTS: Complete response (CR) was achieved in 2 of the 3 patients treated with PUVA (they had stage IB-MF). Both had relapsed after 40 sessions and 7 months later, respectively. Relapses were treated by PUVA maintenance treatment (2 sessions/2 weeks) and NBUVB leading to CR. The third patient (stage IIA) did not respond to PUVA. CR was obtained in the 5 patients treated with NBUVB (stage IA-IB), after a mean cumulative dose of 58.3 J/cm(2). No relapse was noted with NBUVB after a mean follow-up period of 13.2 months (extremes: 2 -31 months). CONCLUSION: NBUVB is only effective on non infiltrated plaques independently of phototype. PUVA therapy is effective even on infiltrated plaques. Both treatments do not prevent relapses. PMID- 20025843 TI - [Migraine or headache management: a pharmacy survey]. AB - AIM: To describe the characteristics and the management of migraine. METHOD: Data on headaches, drug consumption and life habits of 762 patients were collected using questionnaires and pharmacy records. RESULTS: The migraine attack was characterized by a severe pain for more than 80% of the patients. The frequency was more than 2 attacks a week in 16% of the cases. Eighty four per cent of the patients had triptans and 45% had a long-term migraine treatment. Nonspecific analgesics were prescribed for 55%. The frequency of over-consumption of treatments of migraine attacks was 46%. CONCLUSION: The management of migraine still remains inadequate. The pharmacist could contribute to its improvement. PMID- 20025844 TI - Immunohistochemical study of the vaginal inflammatory response in experimental trichomoniasis. AB - In the present paper, the acute and subchronical inflammatory processes of the vaginal epithelial were studied in mice experimentally infected with two Trichomonas vaginalis strains of different pathogenicity, by means of histological and immunological methods. There was an increase in the stratified epithelium layers as well as edema produced by the increase of vascularization in the propia submucosa and infiltration of leukocytes. The proliferation of the vaginal epithelium favors the settlement and persistence of the parasitic infection. All of the findings corresponded with signs of a systemic disease being observed in the animals, including significant weight loss and also intestinal invasion. The entire inflammatory process has been corroborated by studies of adhesion molecules such as E-Selectin, VCAM-1 and PECAM-1. A correlation between the time of appearance and the perseverance of the inflammatory process with E-Selectin and VCAM-1 expression was observed, but not with PECAM-1. The strain with a higher pathogenicity was able to invade deep vaginal tissues and thus, parasites could not be detected by vaginal washings. This may be an important cause of diagnosis and treatment failure. Also, by the different localization of trichomonads, it appeared that the battle between host and parasite took place in different areas dependent upon the characteristics of the strain. PMID- 20025845 TI - Selective binding of the fluorescent dye 1-anilinonaphthalene-8-sulfonic acid to peroxisome proliferator-activated receptor gamma allows ligand identification and characterization. AB - Peroxisome proliferator-activated receptor gamma (PPARgamma) is a member of the nuclear receptor superfamily involved in insulin sensitization, atherosclerosis, inflammation, and carcinogenesis. PPARgamma transcriptional activity is modulated by specific ligands that promote conformational changes allowing interaction with coactivators. Here we show that the fluorophore 1-anilinonaphthalene-8-sulfonic acid (ANS) binds to PPARgamma-LBD (ligand binding domain), displaying negligible interaction with other nuclear receptors such as PPARalpha and retinoid X receptor alpha (RXRalpha). ANS binding is competed by PPARgamma agonists such as rosiglitazone, 15-deoxy-Delta(12,14)-prostaglandin J(2) (15d-PGJ(2)), and 9,10 dihydro-15-deoxy-Delta(12,14)-prostaglandin J(2) (CAY10410). Moreover, the affinity of PPARgamma for these ligands, determined through ANS competition titrations, is within the range of that reported previously, thereby suggesting that ANS competition could be useful in the screening and characterization of novel PPARgamma agonists. In contrast, gel-based competition assays showed limited performance with noncovalently bound ligands. We applied the ANS binding assay to characterize a biotinylated analog of 15d-PGJ(2) that does not activate PPAR in cells. We found that although this compound bound to PPARgamma with low affinity, it failed to promote PPARgamma interaction with a fluorescent SRC-1 peptide, indicating a lack of receptor activation. Therefore, combined approaches using ANS and fluorescent coactivator peptides to monitor PPARgamma binding and interactions may provide valuable strategies to fully understand the role of PPARgamma ligands. PMID- 20025847 TI - Visualizing association of lipidated signaling proteins in heterogeneous membranes--partitioning into subdomains, lipid sorting, interfacial adsorption, and protein association. AB - In a combined chemical biological and biophysical approach, we studied the partitioning of differently fluorescent-labeled palmitoyl and/or farnesyl lipidated peptides, which represent membrane recognition model systems, as well as the full lipidated N-Ras protein into various model membrane systems including canonical model raft mixtures. To this end, two-photon fluorescence microscopy on giant unilamellar vesicles, complemented by tapping-mode atomic force microscopy (AFM) measurements, was carried out. The measurements were performed over a wide temperature range, ranging from 30 to 80 degrees C to cover different lipid phase states (solid-ordered (gel), fluid/gel, liquid-ordered/liquid-disordered, all fluid). The results provide direct evidence that partitioning of the lipidated peptides and N-Ras occurs preferentially into liquid-disordered lipid domains, which is also reflected in a faster kinetics of incorporation. The phase sequence of preferential binding of N-Ras to mixed-domain lipid vesicles is liquid disordered>liquid-ordered>>solid-ordered. Intriguingly, we detect - using the better spatial resolution of AFM - also a large proportion of the lipidated protein located at the liquid-disordered/liquid-ordered phase boundary, thus leading to a favorable decrease in line tension that is associated with the rim of neighboring domains. In an all-liquid-ordered, cholesterol-rich phase, phase separation can be induced by an effective lipid sorting mechanism owing to the high affinity of the lipidated peptides and proteins to a fluid-like lipid environment. At low temperatures, where the overall acyl chain order parameter of the lipid bilayer has markedly increased, such an efficient lipid sorting mechanism is energetically too costly and self-association of the peptide into small clusters takes place. These data reveal the interesting ability of the lipidated peptides and proteins to induce formation of fluid microdomains at physiologically relevant high cholesterol concentrations. Furthermore, our results reveal self-association of the N-Ras protein at the domain boundaries which may serve as an important vehicle for association processes and nanoclustering, which has also been observed in in vivo studies. PMID- 20025846 TI - Ascochlorin is a novel, specific inhibitor of the mitochondrial cytochrome bc1 complex. AB - Ascochlorin is an isoprenoid antibiotic that is produced by the phytopathogenic fungus Ascochyta viciae. Similar to ascofuranone, which specifically inhibits trypanosome alternative oxidase by acting at the ubiquinol binding domain, ascochlorin is also structurally related to ubiquinol. When added to the mitochondrial preparations isolated from rat liver, or the yeast Pichia (Hansenula) anomala, ascochlorin inhibited the electron transport via CoQ in a fashion comparable to antimycin A and stigmatellin, indicating that this antibiotic acted on the cytochrome bc(1) complex. In contrast to ascochlorin, ascofuranone had much less inhibition on the same activities. On the one hand, like the Q(i) site inhibitors antimycin A and funiculosin, ascochlorin induced in H. anomala the expression of nuclear-encoded alternative oxidase gene much more strongly than the Q(o) site inhibitors tested. On the other hand, it suppressed the reduction of cytochrome b and the generation of superoxide anion in the presence of antimycin A(3) in a fashion similar to the Q(o) site inhibitor myxothiazol. These results suggested that ascochlorin might act at both the Q(i) and the Q(o) sites of the fungal cytochrome bc(1) complex. Indeed, the altered electron paramagnetic resonance (EPR) lineshape of the Rieske iron-sulfur protein, and the light-induced, time-resolved cytochrome b and c reduction kinetics of Rhodobacter capsulatus cytochrome bc(1) complex in the presence of ascochlorin demonstrated that this inhibitor can bind to both the Q(o) and Q(i) sites of the bacterial enzyme. Additional experiments using purified bovine cytochrome bc(1) complex showed that ascochlorin inhibits reduction of cytochrome b by ubiquinone through both Q(i) and Q(o) sites. Moreover, crystal structure of chicken cytochrome bc(1) complex treated with excess ascochlorin revealed clear electron densities that could be attributed to ascochlorin bound at both the Q(i) and Q(o) sites. Overall findings clearly show that ascochlorin is an unusual cytochrome bc(1) inhibitor that acts at both of the active sites of this enzyme. PMID- 20025848 TI - Dynamics of lipid domain formation: fluctuation analysis. AB - Scanning-fluctuation correlation spectroscopy was used to detect subresolution organizational fluctuations in the lipid liquid-crystalline phase for single lipid model systems. We used the fluorescent probe Laurdan which is sensitive to the amount of water in the membrane to show that there is a spatial heterogeneity on the scale of few pixels (the size of the pixel is 50 nm). We calculated the pixel variance of the GP function and we found that the variance has a peak at the phase transition for 3 different samples made of pure lipids. The pixel variance has an abrupt change at the phase transition of the membrane and then it slowly decreases at higher temperature. The relatively large variance of the GP indicates that the liquid phase of the membrane is quite heterogeneous even several degrees higher than the phase transition temperature. We interpreted this result as evidence of an underlying microscale structure of the membrane in which water is not uniformly distributed at the micron scale. Imaging of these microstructures shows that the pixels with different GP tend to concentrate in specific domains in the membrane. In the case of single lipid membrane, the statistical and fluctuation analysis of the GP data shows that even such simple lipid systems are capable of generating and maintaining stable structural and organizational heterogeneities. PMID- 20025849 TI - Moxifloxacin increases anti-tumor and anti-angiogenic activity of irinotecan in human xenograft tumors. AB - Camptothecins (CPTs) are topoisomerase I inhibitors chemotherapeutic agents used in combination chemotherapy. We showed previously that combination of moxifloxacin (MXF) and CPT induced inhibitory effects on topoisomerase I activity, on proliferation of HT-29 cells in vitro and enhanced apoptosis, compared to CPT alone. Analysis of secretion of the pro-angiogenic factors IL-8 and VEGF showed significant reduction by MXF. Using a murine model of human colon carcinoma xenograft, we compared the effects of MXF/CPT in vitro to MXF/irinotecan combination in vivo. We show that the MXF/CPT inhibitory effects observed in vitro are reflected in the inhibition of the progressive growth of HT 29 cells implanted in SCID mice. Using caliper measurements, Doppler ultrasonography, image analyses and immunohistochemistry of nuclear proteins (Ki 67) and vascular endothelial cells (CD-31) we show that addition of MXF (45mg/kg) to a relatively ineffective dose of irinotecan (20mg/kg), results in a 50% and 30% decrease, respectively, in tumor size and a decrease in Ki-67 staining. Power Doppler Ultrasound showed a significant, pronounced decrease in the number of blood vessels, as did CD-31 staining, indicating decreased blood flow in tumors in mice treated with MXF alone or MXF/irinotecan compared to irinotecan. These results suggest that the combination of MXF/irinotecan may result in enhanced anti-neoplastic/anti-angiogenic activity. PMID- 20025850 TI - Coordinate regulation/localization of the carbohydrate responsive binding protein (ChREBP) by two nuclear export signal sites: discovery of a new leucine-rich nuclear export signal site. AB - Carbohydrate response element binding protein (ChREBP) is responsible for conversion of dietary carbohydrate to storage fat in liver by coordinating expression of the enzymes that channel glycolytic pyruvate into lipogenesis. The activation of ChREBP in response to high glucose is nuclear localization and transcription, and the inactivation of ChREBP under low glucose involves export from the nucleus to the cytosol. Here we report a new nuclear export signal site ("NES1") of ChREBP. Together these signals provide ChREBP with two NES sequences, both the previously reported NES2 and now the new NES1 coordinate to interact together with CRM1 (exportin) for nuclear export of the carbohydrate response element binding protein. PMID- 20025851 TI - The effect of smoking on pain-related evoked potentials. AB - The effects of human tobacco smoking and nicotine on pain-related brain activities were investigated. EEG responses evoked by a painful laser beam (laser evoked potentials; LEPs), and the plasma nicotine concentration (PNC) were measured. There were two sessions, one after smoking (Smoking session), and the other in no smoking (Control session). Subjective ratings of pain perception were also measured using the visual analog scale (VAS). Two major components, N2 and P2 of LEPs, were recorded. The amplitude of P2 was significantly smaller in the Smoking session than in the Control session. A significant negative correlation was found between PNC and the amplitude of N2 as well as P2. The results were consistent with the hypothesis that smoking and/or nicotine have an antinociceptive effect, which supports most non-human studies and some human studies. Smoking of a single tobacco cigarette did not show a subjectively perceivable extent of reduction in the intensity of evoked pain. PMID- 20025852 TI - Effects of lamotrigine and topiramate on hippocampal neurogenesis in experimental temporal-lobe epilepsy. AB - Lamotrigine (LTG) and topiramate (TPM), two of the most commonly used new generation antiepileptic drugs (AEDs), have been shown to produce no adverse and impaired cognitive effects in patients with epilepsy, respectively. As seizure induced neurogenesis might contribute to cognitive deficits that are associated with status epilepticus (SE), we examined whether these two drugs produce differential effects on seizure-induced neurogenesis in the hippocampus of adult rats. Lithium pilocarpine model was used to mimic human temporal-lobe epilepsy. Five hours after SE, LTG and TPM were administered intragastrically twice daily throughout the entire length of the experiment with total daily dose of 20 and 80 mg/kg, respectively. The hippocampal neurogenesis was examined using 5 bromodeoxyuridine and doublecortin immunohistochemistry. Both LTG and TPM treatments significantly inhibited seizure-induced proliferation of neural progenitors in the hippocampus, but did not affect the neuronal differentiation of newborn cells. Long-term treatment with both AEDs decreased the number of spontaneous recurrent seizures after SE and alleviated chronic seizure-induced neuronal injury in the dentate hilus. Eventually, TPM significantly increased the number of newborn neurons in the dentate granular cell layer after seizures likely by promoting the survival of newborn neurons. In contrast, LTG treatment significantly reduced the number of ectopic hilar newborn neurons after seizures. Neither of them prevented the formation of hilar basal dendrites of newborn neurons in the epileptic hippocampus. These results indicate that TPM but not LTG promotes aberrant neuron regeneration in the hippocampus after SE, which might be partially related to their differential effects on cognitive function. PMID- 20025853 TI - Acupuncture needling sensation: the neural correlates of deqi using fMRI. AB - The needling sensation of deqi is considered by most acupuncturists to be an important component of acupuncture, yet neuroimaging research that investigates this needle sensation has been limited. In this study we have investigated the effect of deqi and acute pain needling sensations upon brain fMRI blood oxygen level-dependent (BOLD) signals. Seventeen right-handed participants who received acupuncture at the right LI-4 (Hegu) acupoint were imaged in a 3T MRI scanner. fMRI datasets were classified, on the basis of psychophysical participants' reports of needling scores, into those that were associated with predominantly deqi sensations versus those with predominantly acute pain sensations. Brain areas showing changes in BOLD signal increases (activations) and decreases (deactivations) were identified. Differences were demonstrated in the pattern of activations and deactivations between groupings of scans associated with deqi versus pain sensations. For the deqi grouping, significant deactivations occurred, whereas significant activations did not. In contrast, the predominantly acute pain grouping was associated with a mixture of activations and deactivations. For the comparison between the predominately deqi sensation grouping and the acute pain sensation grouping (deqi>pain contrast), only negative Z value voxels resulted (mainly from deactivations in the deqi grouping and activations in the pain grouping) in the limbic/sub-cortical structures and the cerebellum regions of interest. Our results show the importance of collecting and accounting for needle sensation data in neuroimaging studies of acupuncture. PMID- 20025854 TI - Neuroprotective effects of (-)-epigallocatechin-3-gallate against quinolinic acid induced excitotoxicity via PI3K pathway and NO inhibition. AB - Excessive stimulation of the NMDA receptor induces neuronal cell death and is implicated in the development of several neurodegenerative diseases. While EGCG suppresses apoptosis induced by NMDA receptor-mediated excitotoxicity, the mechanisms underlying this process have yet to be completely determined. This study was designed to investigate whether (-)-epigallocatechin-3-gallate (EGCG) plays a neuroprotective role by inhibiting nitric oxide (NO) production and activating cellular signaling mechanisms including MAP kinase, PI3K, and GSK 3beta and acting on the antiapoptotic and the proapoptotic genes in N18D3 neural cells. The cells were pretreated with EGCG for 2 h and then exposed to quinolinic acid (QUIN), a NMDA receptor agonist, 30 mM for 24 h. MTT assay and DAPI staining were used to identify cell viability and apoptosis, respectively, and demonstrated that EGCG significantly increased cell viability and protected the cells from apoptotic death. In addition, EGCG had a capacity to reduce QUIN induced excitotoxic cell death not only by blocking increase of intracellular calcium levels but also by inhibiting NO production. Gene expression analysis revealed that EGCG prevented the QUIN-induced expression of the proapoptotic gene, caspase-9, and increased that of the antiapoptotic genes, Bcl-XL, Bcl-2, and Bcl-w. Further examination about potential cell signaling candidate involved in this neuroprotective effect showed that immunoreacitivity of PI3K was significantly increased in the cells treated with EGCG. These results suggest that the neuroprotective mechanism of EGCG against QUIN-induced excitotoxic cell death includes regulation of PI3K and modulation of cell survival and death genes through decreasing of intracellular calcium levels and controlling of NO production. PMID- 20025855 TI - Identification of distinct cellular pools of interleukin-1beta during the evolution of the neuroinflammatory response induced by transient middle cerebral artery occlusion in the brain of rat. AB - The proinflammatory cytokine interleukin(IL)-1beta plays a crucial role in ischemic pathophysiology, since pharmacologic inhibition of its biological effects provides neuroprotection after stroke. However, there is evidence suggesting that under certain circumstances the cytokine may also exert beneficial functions on brain injury. We have investigated the regional and cellular expression of IL-1beta after ischemia-reperfusion injury in the brain of rat, and correlated cytokine expression with the activation/recruitment of glial cells in the damaged tissue. By using a double immunofluorescence histochemical approach, we observed an increased cytokine immunoreactivity in the ischemic core, as early as 1 h after middle cerebral artery occlusion, in few activated OX 42-positive microglial cells and in perivascular GFAP-positive astrocytes, suggesting that the cytokine may participate in the early response of the neurovascular unit to reduced blood supply. After 2 h ischemia, followed by 2 h reperfusion, cytokine staining was evident in the astrocytes of the penumbra and in activated microglial cells of the ischemic core. Microglial activation increases with the progression of damage and, after 22 h reperfusion, OX-42 immunopositive cells were strongly labelled for IL-1beta in the core and, even more intensely, in the penumbra. At this later stage, GFAP-positive cells, appearing hypertrophic and distributed in a ring-like pattern around the ischemic core, do no longer express IL-1beta. Thus, a specific cellular and regional pattern of IL-1beta expression characterises the progression of ischemia reperfusion injury. Depending on the stage and intensity of the insult, the different cellular origin of the cytokine may suggest a distinct role of this neuroinflammatory mediator in ischemic pathophysiology. PMID- 20025856 TI - Amygdala activity in response to forward versus backward dynamic facial expressions. AB - Observations of dynamic facial expressions of emotion activate several brain regions, but the psychological functions of these regions remain unknown. To investigate this issue, we presented dynamic facial expressions of fear and happiness forwards or backwards, thus altering the emotional meaning of the facial expression while maintaining comparable visual properties. Thirteen subjects passively viewed the stimuli while being scanned using fMRI. After image acquisition, the subject's emotions while perceiving the stimuli were investigated using valence and intensity scales. The left amygdala showed higher activity in response to forward compared with backward presentations, for both fearful and happy expressions. Amygdala activity showed a positive relationship with the intensity of the emotion experienced. These results suggest that the amygdala is not involved in the visual but is involved in the emotional processing of dynamic facial expressions, including specifically the elicitation of subjective emotions. PMID- 20025857 TI - Modulation of the benzene metabolite hydroquinone induced toxicity: evidence for an important role of fau. PMID- 20025858 TI - Kinetics of the lamellar gel-fluid transition in phosphatidylcholine membranes in the presence of sugars. AB - Phase diagrams are presented for dipalmitoylphosphatidylcholine (DPPC) in the presence of sugars (sucrose) over a wide range of relative humidities (RHs). The phase information presented here, determined by small angle X-ray scattering (SAXS), is shown to be consistent with previous results achieved by differential scanning calorimetry (DSC). Both techniques show a significant effect of sucrose concentration on the phase behaviour of this phospholipid bilayer. An experimental investigation into the effect of sugars on the kinetic behaviour of the gel to fluid transition is also presented showing that increasing the sugar content appears to slightly increase the rate at which the transition occurs. PMID- 20025859 TI - Location of the mutation site in the first two reported cases of analbuminemia. AB - OBJECTIVE: To find the mutation causing the first two reported cases of analbuminemia. DESIGN AND METHODS: DNA was extracted from blocks of fixed embedded liver. Exons of the albumin gene were amplified and sequenced. RESULTS: A substitution of C>T in exon 12 was found in both subjects, changing the codon CGA for arginine (aa509) to TGA, a stop codon. CONCLUSIONS: The data suggest that analbuminemia is the result of widely scattered random mutations. PMID- 20025860 TI - Clinical and biochemical characteristics of patients with urea cycle disorders in a developing country. AB - OBJECTIVES: To report the clinical and laboratory characteristics of urea cycle disorder (UCD) patients at a tertiary care center in a developing country. DESIGN AND METHODS: Retrospective study of clinical and laboratory data of UCD patients. RESULTS: Thirty-seven UCD patients were studied, 31 symptomatic (high risk) patients (15 neonatal onset, 16 late onset) and 6 with positive neonatal screening. Argininosuccinate synthetase deficiency was the most frequent disease (17/37, 46%), followed by ornithine transcarbamylase (10/37, 27%), arginase (7/37, 19%), and argininosuccinate lyase (3/37, 8%) deficiencies. Mortality of symptomatic patients was 38% (10/26), neonatal onset had the worst outcome, with 50% of survival. CONCLUSIONS: In Mexico, the mortality of the UCD patients is higher than those reported in other countries, and neurological sequels are frequent and severe. It is essential to implement practice guidelines for the professional management of these patients. PMID- 20025861 TI - Neurog3 gene dosage regulates allocation of endocrine and exocrine cell fates in the developing mouse pancreas. AB - The basic helix-loop-helix transcription factor Neurog3 (Neurogenin3 or Ngn3) actively drives endodermal progenitor cells towards endocrine islet cell differentiation during embryogenesis. Here, we manipulate Neurog3 expression levels in endocrine progenitor cells without altering its expression pattern using heterozygosity and a hypomorph. Lowered Neurog3 gene dosage in the developing pancreatic epithelium reduces the overall production of endocrine islet cells without significantly affecting the proportions of various islet cell types that do form. A reduced Neurog3 production level in the endocrine-directed pancreatic progenitor population activates the expression of Neurog3 in an increased number of epithelial progenitors. Yet a significant number of these Neurog3+ cells detected in heterozygous and hypomorphic pancreata, possibly those that express low levels of Neurog3, move on to adopt pancreatic ductal or acinar fates. These data directly demonstrate that achieving high levels of Neurog3 expression is a critical step for endocrine commitment from multipotent pancreatic progenitors. These findings also suggest that a high level of Neurog3 expression could mediate lateral inhibition or other unknown feedback mechanisms to regulate the number of cells that initiate Neurog3 transcription and protein production. The control of Neurog3+ cell number and the Neurog3 threshold dependent endocrine differentiation mechanism combine to select a specific proportion of pancreatic progenitor cells to adopt the islet cell fate. PMID- 20025862 TI - Disruption of Paneth and goblet cell homeostasis and increased endoplasmic reticulum stress in Agr2-/- mice. AB - Anterior Gradient 2 (AGR2) is a protein disulfide isomerase that plays important roles in diverse processes in multiple cell lineages as a developmental regulator, survival factor and susceptibility gene for inflammatory bowel disease. Here, we show using germline and inducible Agr2-/- mice that Agr2 plays important roles in intestinal homeostasis. Agr2-/- intestine has decreased goblet cell Mucin 2, dramatic expansion of the Paneth cell compartment, abnormal Paneth cell localization, elevated endoplasmic reticulum (ER) stress, severe terminal ileitis and colitis. Cell culture experiments show that Agr2 expression is induced by ER stress, and that siRNA knockdown of Agr2 increases ER stress response. These studies implicate Agr2 in intestinal homeostasis and ER stress and suggest a role in the etiology of inflammatory bowel disease. PMID- 20025863 TI - Enhancers: the abundance and function of regulatory sequences beyond promoters. AB - Transcriptional control in mammals and Drosophila is often mediated by regulatory sequences located far from gene promoters. Different classes of such elements - particularly enhancers, but also locus control regions and insulators - have been defined by specific functional assays, although it is not always clear how these assays relate to the function of these elements within their native loci. Recent advances in genomics suggest, however, that such elements are highly abundant within the genome and may represent the primary mechanism by which cell- and developmental-specific gene expression is accomplished. In this review, we discuss the functional parameters of enhancers as defined by specific assays, along with the frequency with which they occur in the genome. In addition, we examine the available evidence for the mechanism by which such elements communicate or interact with the promoters they regulate. PMID- 20025864 TI - Genetic fate mapping demonstrates contribution of epicardium-derived cells to the annulus fibrosis of the mammalian heart. AB - The annulus fibrosis electrically insulates the atria and ventricles, allowing the timed sequential beating of these structures that is necessary for efficient heart function. Abnormal development of the annulus fibrosis leads to persistence of accessory electrical pathways from atria to ventricles, providing the anatomical substrate for re-entrant cardiac arrhythmias such as Wolff-Parkinson White syndrome. To better understand the development of the annulus fibrosis and the etiology of these cardiac arrhythmias, we used Cre-LoxP technology to assess the contribution of epicardium derived cells (EPDCs) to the annulus fibrosis. We found that EPDCs migrated into the region of the forming annulus fibrosis, marked by the protein periostin. These EPDCs also stained positive for procollagen I, suggesting that the EPDCs themselves synthesize proteins of the annulus fibrosis. To further test the hypothesis that EPDCs contribute to cells that synthesize the annulus fibrosis, we purified genetically marked EPDCs from the atrioventricular region and measured gene expression by quantitative PCR. These EPDCs were highly enriched for mRNAs encoding periostin, procollagen I, fibronectin I, vimentin, discoidin domain receptor 2, and tenascin C, markers of fibroblasts and components of the annulus fibrosis. In addition, these EPDCs were highly enriched for Snail, Smad1, Slug, and Twist1, markers for epithelial-to-mesenchymal transition (EMT), and a metalloprotease, Mmp2, that contributes to cellular migration. Our work provides for the first time definitive evidence that epicardium contributes to formation of the mammalian annulus fibrosis through EMT. Abnormalities of this differentiation process may underlie development of some forms of re-entrant atrioventricular tachycardia. PMID- 20025865 TI - Sox2 is required for maintenance and regeneration, but not initial development, of hair cells in the zebrafish inner ear. AB - Sox2 has been variously implicated in maintenance of pluripotent stem cells or, alternatively, early stages of cell differentiation, depending on context. In the developing inner ear, Sox2 initially marks all cells in the nascent sensory epithelium and, in mouse, is required for sensory epithelium formation. Sox2 is eventually downregulated in hair cells but is maintained in support cells, the functional significance of which is unknown. Here we describe regulation and function of sox2 in the zebrafish inner ear. Expression of sox2 begins after the onset of sensory epithelium development and is regulated by Atoh1a/b, Fgf and Notch. Knockdown of sox2 does not prevent hair cell production, but the rate of accumulation is reduced due to sporadic death of differentiated hair cells. We next tested the capacity for hair cell regeneration following laser ablation of mature brn3c:gfp-labeled hair cells. In control embryos, regeneration of lost hair cells begins by 12 h post-ablation and involves transdifferentiation of support cells rather than asymmetric cell division. In contrast, regeneration does not occur in sox2-depleted embryos. These data show that zebrafish sox2 is required for hair cell survival, as well as for transdifferentiation of support cells into hair cells during regeneration. PMID- 20025866 TI - Gamma-protocadherins regulate the functional integrity of hypothalamic feeding circuitry in mice. AB - The hypothalamic neuronal circuits that modulate energy homeostasis become mature and functional during early postnatal life. However, the molecular mechanism underlying this developmental process remains largely unknown. Here we use a mouse genetic approach to investigate the role of gamma-protocadherins (Pcdh gammas) in hypothalamic neuronal circuits. First, we show that rat insulin promoter (RIP)-Cre conditional knockout mice lacking Pcdh-gammas in a broad subset of hypothalamic neurons are obese and hyperphagic. Second, specific deletion of Pcdh-gammas in anorexigenic proopiomelanocortin (POMC) expressing neurons also leads to obesity. Using cell lineage tracing, we show that POMC and RIP-Cre expressing neurons do not overlap but interact with each other in the hypothalamus. Moreover, excitatory synaptic inputs are reduced in Pcdh-gamma deficient POMC neurons. Genetic evidence from both knockout models shows that Pcdh-gammas can regulate POMC neuronal function autonomously and non-autonomously through cell-cell interaction. Taken together, our data demonstrate that Pcdh gammas regulate the formation and functional integrity of hypothalamic feeding circuitry in mice. PMID- 20025867 TI - CXCR4 positive bone mesenchymal stem cells migrate to human endothelial cell stimulated by ox-LDL via SDF-1alpha/CXCR4 signaling axis. AB - BACKGROUND: Bone mesenchymal stem cells (BMSCs) are attractive candidates for cell based therapies to cardiovascular disease such as infarction and atherosclerosis; however, the mechanisms responsible for stem cell chemotaxis and homing remain unknown. Chemokine stromal cell-derived factor 1 (SDF-1alpha) is involved in the process of atherogenesis. This study was aimed at investigating whether the SDF-1alpha of human umbilical vein endothelial cells (HUVECs) plays a role in migration of BM-derived CXCR4(+)(receptor for SDF-1alpha) stem cells. METHODS: HUVECs were cultured from human umbilical cords and was treated with ox LDL. The mRNA and protein expression of SDF-1alpha was detected in HUVECs. CXCR4(+)BMSCs from bone marrow were isolated and were tested by migration and adhesion assays. RESULTS: It was found that ox-LDL induced HUVECs to increase the mRNA and protein expression of SDF-1alpha. Ox-LDL increased the migratory and adhesion response of CXCR4(+)BMSCs. When the neutralizing SDF-1alpha antibody abrogated the secreted SDF-1alpha, the migration and adhesion response of CXCR4(+)BMSCs markedly decreased. CONCLUSIONS: Our data indicated that the endothelial cells (ECs) stimulated by ox-LDL could increase the BMSCs migratory response via SDF-1alpha/CXCR4 signaling axis. These findings provide a new paradigm for biological effects of ox-LDL and have implications for novel stem cell therapeutic strategies for atherosclerosis. PMID- 20025868 TI - Protein and gene expression of estrogen receptor alpha and nuclear morphology of two breast cancer cell lines after different fixation methods. AB - We assessed morphology and ERalpha protein and gene expression of two breast cancer cell lines after three different fixatives: neutral-buffered 10% formaldehyde, LN-FIX and FineFIX and varying fixation times. We found that the cell morphology was best preserved in cells fixed with LN-FIX. Two commercial fixatives used in this study shrank cells less than formalin. In immunohistochemical assay samples were stained with two different ERalpha antibodies, clone 1D5 and clone SP1. All tested fixatives were suitable for immunohistochemistry. Staining was more intensive and the number of stained cells was larger with the clone 1D5 than with the clone SP1. Our gene expression analysis showed that formalin and LN-FIX preserve the ERalpha better than FineFIX, which is advertised to be optimal for molecular analysis. Our study suggests that tissues fixed with formalin are suitable also for molecular biology assays. This makes possible to research formalin-fixed paraffin-embedded archival tissues also with molecular techniques. PMID- 20025869 TI - RIP2: a novel player in the regulation of keratinocyte proliferation and cutaneous wound repair? AB - We could recently demonstrate an important role of receptor interacting protein 4 (RIP4) in the regulation of keratinocyte differentiation. Now, we analyzed a potential role of the RIP4 homolog RIP2 in keratinocytes. Specifically, we demonstrate here that rip2 expression is induced by scratch-wounding and after the induction of differentiation in these cells. Furthermore, serum growth factors and cytokines can induce rip2, with TNF-alpha-dependent induction being dependent on p38 MAPK. In addition, we demonstrate that scratch-induced upregulation of rip2 expression is completely blocked by the steroid dexamethasone. Since we also show that RIP2 is an important player in the regulation of keratinocyte proliferation, these data suggest that inhibition of rip2 upregulation after wounding might contribute to the reduced and delayed wound re-epithelialization phenotype seen in glucocorticoid-treated patients. PMID- 20025870 TI - Induction of ANGPTL4 expression in human airway smooth muscle cells by PMA through activation of PKC and MAPK pathways. AB - In this study, we demonstrate that protein kinase C (PKC) activators, including phorbol-12-myristate-13-acetate (PMA), 1,2-dioctanoyl-sn-glycerol (DOG), and platelet-derived growth factor alpha are potent inducers of angiopoietin-like protein 4 (ANGPTL4) expression in several normal lung cell types and carcinoma cell lines. In human airway smooth muscle (HASM) cells induction of ANGPTL4 expression is observed as early as 2 h after the addition of PMA. PMA also increases the level of ANGPTL4 protein released in the medium. PKC inhibitors Ro31-8820 and Go6983 greatly inhibit the induction of ANGPTL4 mRNA by PMA suggesting that this up-regulation involves activation of PKC. Knockdown of several PKCs by corresponding siRNAs suggest a role for PKCalpha. PMA does not activate MAPK p38 and p38 inhibitors have little effect on the induction of ANGPTL4 indicating that p38 is not involved in the regulation of ANGPTL4 by PMA. In contrast, treatment of HASM by PMA induces phosphorylation and activation of Ra, MEK1/2, ERK1/2, JNK, Elk-1, and c-Jun. The Ras inhibitor manumycin A, the MEK1/2 inhibitor U0126, and the JNK inhibitor SP600125, greatly reduce the increase in ANGPTL4 expression by PMA. Knockdown of MEK1/2 and JNK1/2 expression by corresponding siRNAs inhibits the induction of ANGPTL4. Our observations suggest that the induction of ANGPTL4 by PMA in HASM involves the activation of PKC, ERK, and JNK pathways. This induction may play a role in tissue remodeling during lung injury and be implicated in several lung pathologies. PMID- 20025871 TI - Spotlight on...Giorgio Semenza. Interview by Daniela Ruffell. PMID- 20025872 TI - Characterization of corneal keratocyte morphology and mechanical activity within 3-D collagen matrices. AB - The purpose of this study was to assess quantitatively the differences in morphology, cytoskeletal organization and mechanical behavior between quiescent corneal keratocytes and activated fibroblasts in a 3-D culture model. Primary cultures of rabbit corneal keratocytes and fibroblasts were plated inside type I collagen matrices in serum-free media or 10% FBS, and allowed to spread for 1-5 days. Following F-actin labeling using phalloidin, and immunolabeling of tubulin, alpha-smooth muscle actin or connexin 43, fluorescent and reflected light (for collagen fibrils) 3-D optical section images were acquired using laser confocal microscopy. In other experiments, dynamic imaging was performed using differential interference contrast microscopy, and finite element modeling was used to map ECM deformations. Corneal keratocytes developed a stellate morphology with numerous cell processes that ran a tortuous path between and along collagen fibrils without any apparent impact on their alignment. Fibroblasts on the other hand, had a more bipolar morphology with pseudopodial processes (P T transition (43/125; 34% of base changes in the coding region). G-->T transversions occurred at a frequency of 10% (12/125), which is less than reported in mutation databases for head and neck squamous cell carcinoma (24%). Characteristically, in our SNC series, the mutations were scattered over a large number of codons, codon 248 being the most frequent target of base substitution. Codon 135 was the second most frequently mutated codon; this nucleotide position has not been reported before as frequently mutated in head and neck cancer or human cancer in general. About half of all tumours with TP53 mutations carried more than one mutation. Interestingly, 86% (19/22) of the silent mutations detected had occurred in tumours with multiple mutations. PMID- 20025892 TI - Facilitation of bottom-up feature detection following rTMS-interference of the right parietal cortex. AB - In visual search tasks the optimal strategy should utilize relevant information ignoring irrelevant one. When the information at the feature and object levels are in conflict, un-necessary processing at higher level of object shape can interfere with detection of lower level orientation feature. We explored the effects of inhibitory trains of transcranial magnetic stimulation (rTMS) on the right and left parietal cortex in healthy subjects performing two visual search tasks. One task (Task A) was characterised by an object-to-feature interference. The other task (Task B) was without such interference. We found that rTMS of the right parietal cortex significantly reduced reaction times (RTs) in Task A, where object recognition interferes with detection of orientation. This significant RT reduction was present only for the first 10 trials. Interestingly, right parietal rTMS had no effect on Task B. Moreover, rTMS of the left parietal cortex did not modify subjects' RTs in either task. Subjects' accuracy was equally affected by rTMS in both tasks over time. We suggest that inhibition of the right parietal cortex by means of rTMS facilitates feature-based visual search by inhibiting the interfering feature binding and spatial attentional processes. This allows subjects to accomplish Task A faster. PMID- 20025893 TI - A new steroidal 5,7-diene derivative, 3beta-hydroxyandrosta-5,7-diene-17beta carboxylic acid, shows potent anti-proliferative activity. AB - The new steroidal 5,7-diene, 3beta-hydroxyandrosta-5,7-diene-17beta-carboxylic acid (17-COOH-7DA), was synthesized from 21-acetoxypregnenolone, with the oxidative cleavage of the side chain being dependent on the presence of oxygen. In human epidermal (HaCaT) keratinocytes, 17-COOH-7DA inhibited proliferation in a dose-dependent manner, starting at a dose as low as 10(-11) M. This inhibition was accompanied by decreased expression of epidermal growth factor receptor, bcl2 and cyclin E2 mRNAs and by increased expression of involucrin mRNA. Inhibition of proliferation was associated with slowing of the cell cycle in G1/G0 phases but not with cell death. 17-COOH-7DA was significantly more potent than pregnenolone, 17-COOH-pregnenolone, 17-COOCH(3)-7DA and calcitriol. 17-COOH-7DA also inhibited proliferation of normal human epidermal melanocytes and human and hamster melanoma lines, however, with lower potency than for keratinocytes. In normal human dermal fibroblasts 17-COOH-7DA stimulated proliferation in serum-free media but inhibited it in the presence of 5% serum. 17-COOH-7DA inhibited cell colony formation of human and hamster melanoma cells, and induced monocyte-like differentiation of human HL60 leukemia cells. Thus, the new steroidal 5,7-diene, 17-COOH-7DA, can serve as an inhibitor of proliferation of normal keratinocytes and normal and malignant melanocytes, as a condition-dependent regulator of fibroblast proliferation and a stimulator of leukemia cell differentiation. PMID- 20025894 TI - Dermatotoxicology: historical perspective and advances. AB - The fundamental principles underlying the study of dermatotoxicology were developed by Arnold Lehman and John Draize over a half century ago and remain applicable today. This discipline has proven indispensable for addressing the problems associated with skin exposure to chemicals. The 55th anniversary of Lehman's landmark publication on safety factors presents the opportunity to reflect upon the historical beginnings of dermatotoxicology and the role of regulatory policies on the development of this field over the years. The complexity and sheer volume of information that has been collected makes it difficult to comprehensively cover all aspects of this vast discipline. This overview will touch upon the general concepts of ADME, the various forms of contact dermatitis, and transdermal drug delivery systems. The traditional tests performed in animals and humans to identify allergic or irritant potential of chemicals, in addition to alternative methods such as QSAR modeling will be discussed. The subspecialties of infant and occupational dermatotoxicology, as well as dermatotoxicology of aged and ethnic skin, and skin of the vulva and vagina will also be noted. PMID- 20025895 TI - The lethality of tentacle-only extract from jellyfish Cyanea capillata is primarily attributed to cardiotoxicity in anaesthetized SD rats. AB - Previous studies in our laboratory have shown that tentacle-only extract (TOE) has similar hypotensive effects with nematocyst venom from jellyfish Cyanea capillata, and the experimental studies on the in vivo cardiovascular effects of TOE were further performed to explore the leading cause of death and analyze the basic physiopathologic change in anaesthztized SD rats. Plots of TOE dose versus time to death showed dose-dependent curvilinear relationship. ECG changed in a dose- and time-dependent manner. Haemodynamic parameters, including the heart rate, mean femoral arterial pressure, left ventricular developed pressure and the first derivative of left ventricular pressures, decreased, but left ventricular end-diastolic pressure did not increase. Arterial partial pressure of oxygen and oxygen saturation did not change. Lactate dehydrogenase, creatine kinase and MB isoenzyme of creatine kinase increased significantly. Histopathological examination showed congestion, haemorrhage, edema and denaturation in the heart; congestion, haemorrhage in the lung and acute congestion in the liver. Transmission electron microscopy examination found that parts of sarcomeric filaments disrupted, dissolved and disappeared, and parts of mitochondria swelled in cardiocytes. Laser scanning confocal microscope examination found that ventricular myocytes from adult rat were deformed and ultimately died within 30 min after TOE treatment. Our results reveal that cardiodepressive effect of C. capillata TOE is the leading cause of death and acute total heart failure is the basic physiopathologic change in anaesthetized SD rats. PMID- 20025896 TI - A search order lost effect: ignoring a singleton distractor affects visual search efficiency. AB - Four experiments investigated after-effects of attentional capture by a target feature singleton distractor. In Experiments 1 and 3, participants searched for an orientation singleton target in a visual display and responded to a reported attribute in the target (a compound search task). On some trials, a singleton distractor with the same orientation as, but a different color from, the target occurred. In the singleton distractor-absent trials reaction times for targets were unchanged irrespective of the number of nontargets. However, on singleton distractor-present trials, target reaction times increased with number of displayed nontargets. Ignoring target-feature singleton distractors induced inefficient visual searches slowed target search, suggesting that targets were searched serially in the presence of a singleton distractor induces inefficient serial search. This result implies that the search order, corresponding to relative item salience, is lost following attentional capture by a singleton distractor. Subsequent experiments explored conditions that might elicit the search order lost effect. It did not occur when task-irrelevant singleton distractors occurred in a compound search task (Experiment 2) or when target feature (Experiment 4) singleton distractors occurred in a simple target detection task. Together, results suggest that the search order lost effect is mediated by dynamic computations involving saliency and feature maps. An explanation of this effect is proposed. PMID- 20025897 TI - New approaches on crystallization under electric fields. AB - This review presents the state of the art in protein crystallization, nucleation and growth under electric fields. Both external and internal applications of Direct Current (DC) and Alternative Current (AC) experiments are discussed. It is shown that competing effects account for the decreased nucleation time and number of crystals observed yielding larger and sometimes better quality crystals. PMID- 20025898 TI - How transcription factors can adjust the gene expression floodgates. AB - The rate of transcription initiation is the main level of quantitative control of gene expression, primarily responsible for the accumulation of mRNAs in the cell. Many, if not all, molecular actors involved in transcription initiation are known but the mechanisms underlying the frequency of initiations, remain elusive. To make the connection between transcription factors and the frequency of transcription initiation, intricated aspects of this complex activity are classified i) depending on whether or not the DNA-bound transcription factors directly activate the commitment to transcription and ii) on the destructive or non-destructive effect of transcription initiation on the stability of promoter complexes. Two possible sources of synergy allowing the combinatorial specificity of transcription factors action are compared, for binding to DNA and for recruiting transcription machineries. Tentative formulations are proposed to discriminate the different micro-reversible modes of DNA binding cooperativity modulating the specificity and dosage of transcription initiation. PMID- 20025899 TI - Antioxidant activity of sulfated polysaccharide fractions extracted from Undaria pinnitafida in vitro. AB - Two sulfated polysaccharide fractions (S1 and S2) were successfully isolated from seaweed Undaia pinnitafida and the chemical characteristics were determined. Antioxidant activities of the polysaccharide fractions were evaluated by assays of various antioxidants in vitro systems, including superoxide anion, 2,2 diphenyl-1-picrylhydrazyl (DPPH), hydroxyl radical-scavenging activity and metal chelating ability. The results showed that the two sulfated polysaccharides contained rhamnose as the major neutral sugar and present high sulfate content (33.99-34.29%). Antioxidant assays suggested that the two sulfated polysaccharide fractions (S1 and S2) possessed good antioxidant properties and had stronger antioxidant abilities than de-sulfated polysaccharides (DS-1 and DS-2). Available data obtained by in vitro models suggested that the correlation between the sulfate content and antioxidant activity was positive. PMID- 20025900 TI - Release of DNA from surfactant complexes induced by 2-hydroxypropyl-beta cyclodextrin. AB - Decompaction of DNA-CTA self-assembled complexes by 2-hydroxypropyl-beta cyclodextrin (2-HP-beta-CD) was studied and the results were compared with beta CD. Different degrees of 2-HP substitution (0.6, 0.8 and 1.0, respectively) were used and the decompaction was successful with all degrees of substitution. Fluorescence microscopy, steady state fluorescence spectroscopy, density and sound velocity measurements, thermal melting and circular dichroism were used. Compared to previous work using alpha- and beta-CD, the fluorescence spectroscopy results showed that the 2-HP-substituted CDs more efficiently released DNA into solution. Furthermore, dissociation of macroscopically phase separated DNA-CTA complexes was achieved upon addition of 2-HP-beta-CD and the results gave strong indications on the non-equilibrium nature of the system. The globule-to-coil transition was not found to proceed through a coexistence region, which seems to be a general phenomenon in DNA decompaction using CDs. PMID- 20025901 TI - Chemical composition analysis of soybean oligosaccharides and its effect on ATPase activities in hyperlipidemic rats. AB - A water-soluble oligosaccharide was isolated from the water extract of soybean, which is widely grown worldwide due to its nutritional benefits of a full range of amino acids and a high protein content. In this study, we analysed chemical composition of soybean oligosaccharides (SBOS) using high performance liquid chromatography (HPLC). The results obtained were compared with the following sugar standards: mannose, galactose, xylose, arabinose, fucose and glucose. Results showed that SBOS was composed of galactose (65.3%), mannose (15.6%), fructose (7.8%) and glucose (8.7%). The pharmacological action of SBOS has been studied in hyperlipidemic rats fed high-fat diet. In hyperlipidemic rats, feeding with SBOS (150 and 300 mg/kg b.w.) exerted a pharmacological effect as assessed by reversal of plasma levels of lactic acid, nitric oxide (NO), vascular endothelial growth factor (VEGF) levels, and heart Na+-K+-ATPase, Ca2+-ATPase and Mg2+-ATPase activities. These results indicated that SBOS may have beneficial effects on the therapy of some heart diseases, such as coronary artery diseases. PMID- 20025902 TI - The attentional blink: past, present, and future of a blind spot in perceptual awareness. AB - A survey of the attention literature reveals the prominence of the attentional blink (AB)--a deficit in reporting the second of two targets when presented in close temporal succession. For two decades, this robust attentional phenomenon has been a major topic in attention research because it is informative about the rate at which stimuli can be encoded into consciously accessible representations. The pace of discovery and theoretical advancement concerning the AB has increased rapidly in the past few years with emphasis on new neurophysiological evidence and computational accounts of attentional processes. In this review we extract the central questions and the main lessons learnt from the past, and subsequently provide important directions for future research. PMID- 20025903 TI - Monocyte HLA-DR expression as predictor of poor outcome in neonates with late onset neonatal sepsis. AB - OBJECTIVES: Down regulation of HLA-DR expression on monocytes has been reported in adult sepsis. The aims of this study were, first to evaluate monocyte HLA-DR expression in late onset neonatal infection and second to investigate the prognostic value of monocyte HLA-DR expression at onset of symptoms for the final outcome. METHODS: Peripheral blood samples were taken from neonates, who were classified into three groups: late onset neonatal sepsis group (n=40); non infective disorders group (n=24) and the control group (n=25). Monocyte expression of HLA-DR was determined by flow cytometry. RESULTS: The percentage of monocytes expressing HLA-DR was lower in neonates with late onset sepsis (p<0.05). Of the 40 septic patients enrolled in the study, 32 survived, while 8 died. The percentage of HLA-DR expressing monocytes was significantly lower in the non-survivor sepsis group (16.6%) compared with that in the survivor sepsis group (45.2%). The optimal cutoff value of HLA-DR for predicting mortality was 30% with 87% sensitivity and 81% specificity. Patients with monocyte HLA-DR expression 98%), and low residual solvents (<50 ppm), due to rapid particle formation from efficient solvent removal provided by the SFEE process. pFlt23K-PLGA nanoparticles were capable of in vitro transfection, significantly reducing secreted VEGF from human lung alveolar epithelial cells (A549) under normoxic and hypoxic conditions. pFlt23K-PLGA nanoparticles did not exhibit cytotoxicity and are of potential value in treating neovascular disorders wherein VEGF levels are elevated. PMID- 20025946 TI - In vitro delivery of anti-breast cancer agents directly via the mammary papilla (nipple). AB - The objective of this study was to investigate, in vitro, the plausibility of a novel method for delivering a combination of anti-breast cancer agents to the breast via the mammary papilla (nipple). Mammary papillae were prepared from freshly excised strips of porcine sow breasts by blunt dissection. Permeation studies were performed using all glass Franz diffusion cells in both upright and lateral position, with drugs examined individually and in combination. Donor phase was comprised of equimolar PD98059, LY294002 and tamoxifen; 2.54x10(-4) mol dissolved in 950 microL fish oil (containing approximately 23% (w/v) eicosapentaenoic acid, EPA), 25 microL DMSO and 25 microL 1,8-cineole. Also, 4 or 10% Cabosil M5P (w/v) was added to thicken the formulation. After 6 h, the papillae were recovered, cleaned, centrifuged and extracted thrice with methanol. Pooled extracts were analysed by reversed-phase HPLC. The significance of the papilla orientation was also investigated. When applied singly and laterally, the amount extracted from the porcine breast tissue for PD98059, LY294002 and tamoxifen were 1.83+/-0.30, 10.67+/-1.78 and 0.74+/-0.19x10(-2) micromol g(-1) respectively; applied simultaneously and laterally, 2.03+/-0.14, 4.86+/-0.47 and 0.22+/-0.04x10(-2) micromol g(-1) respectively. With 4% Cabosil formulation, amount extracted for PD98059 and LY294002 were 5.71+/-0.95 and 9.91+/-0.92x10(-2) micromol g(-1) respectively; with 10% formulation, 2.64+/-0.5 and 3.90+/-0.78x10( 2) micromol g(-1) respectively. Tamoxifen was below its limit of detection in both Cabosil M5P formulations. To conclude, localized passive delivery via the mammary papilla is a plausible non-invasive means of delivering anti-breast cancer drugs directly to the breast, in levels that have previously been shown to markedly inhibit the growth of breast cancer cell lines, in vitro. The amounts deliverable may be influenced by differential interactions with the thickening agent and patient orientation. PMID- 20025947 TI - Inclusion of poorly soluble drugs in highly ordered mesoporous silica nanoparticles. AB - Silica nanoparticles (MSNs) with a highly ordered mesoporous structures (103A) with cubic Im3 m have been synthesized using triblock copolymers with high poly(alkylene oxide) (EO) segments in acid media. The produced nanoparticles displayed large specific surface area (approximately 765 cm(2)/g) with an average particles size of 120 nm. The loading efficiency was assessed by incorporating three major antiepileptic active substances via passive loading and it was found to varying from 17 to 25%. The state of the adsorbed active agents was further analyzed using differential scanning calorimetry (DSC) and X-ray powder diffraction (XRPD). Dissolution studies revealed rapid release profiles within the first 3 h. The viability of 3T3 endothelial cells was not affected in the presence of MSNs indicating negligible cytotoxicity. PMID- 20025948 TI - The interplay of membrane formation and drug release in solution-cast films of polylactide polymers. AB - The interplay of phase inversion and drug release has been studied for films of several biodegradable polylactide polymers cast from solutions containing polymer, solvent, and drug (naproxen). Variables studied included polymer type and concentration, solvent type, and film casting conditions (i.e. free or forced convection, humidity). Film morphologies and thermal properties indicate that reduction of the T(g) of the amorphous poly (lactide-co-glycolide) (PLGA) and poly (d, l-lactide) (PDLLA) systems caused by the drug, inhibits stabilization of a porous, structure, regardless of dry casting conditions and drug loads. Porous membranes could be formed by wet casting; however, drug loss during casting, makes this a non-viable process. For semi-crystalline PLLA, membrane morphologies could be varied by controlling the mass transfer path to form a single-phase dense film by polymer crystallization or a liquid-liquid two-phase structure followed by locking-in by polymer crystallization. However, the lack of drug solubility in the crystalline phase leads to unfavorable drug distributions most often leading to a burst release. Release profiles for all three polymers were found to follow a two-stage release model, with a first stage diffusive release followed by zero-order release in the second stage due to polymer erosion. PMID- 20025949 TI - Impact of amino acid replacements on in vitro permeation enhancement and cytotoxicity of the intestinal absorption promoter, melittin. AB - Melittin is an amphipathic alpha-helical peptide known to cause the non-cell selective perturbation of cell membranes, especially erythrocytes. The well characterised interaction of the peptide with phospholipid bilayers has led to its use as a model to study lipid-peptide interactions. In recent years, melittin has emerged as a potential intestinal absorption promoter that increases paracellular marker permeability across both in vitro and in situ intestinal drug delivery models. Like many other promoters, inherent toxicity limits the drug delivery potential of melittin. The purpose of this study was to examine the effect of amino acid modifications of melittin on viability and drug permeation in human intestinal epithelial cell monolayers (Caco-2), where each structural change made to the peptide is known to reduce the cytolytic action of the peptide on cell membranes composed of zwitterionic phospholipids. Each of the 4 peptide analogues (PA) demonstrated reduced cytotoxicity in the methylthiazolyldiphenyl tetrazolium bromide (MTT) conversion assay and lactate dehydrogenase (LDH) membrane integrity assay, which was correlated with a reduction in amphipathicity and hydrophobicity, as measured by RP-HPLC. The selected amino acid changes however, also attenuated the epithelial permeation enhancement activity of melittin, as measured by transepithelial electrical resistance (TEER) and flux of FITC-dextran-4 kDa across Caco-2 monolayers. This data suggests that the cytolytic action of melittin is responsible in part for permeation enhancement and that these effects are related to transcellular perturbation in addition to effects on tight junctions. PMID- 20025950 TI - Transdermal iontophoretic delivery of terbinafine hydrochloride: quantitation of drug levels in stratum corneum and underlying skin. AB - The objective of this study was to determine the effect of iontophoresis on the delivery of terbinafine hydrochloride (4%, w/w) into and across hairless rat skin. In vitro skin uptake and permeation studies were performed using Franz diffusion cells. Anodal iontophoresis was applied for 1h at current densities of 0.2, 0.3 and 0.4mA/cm(2). In addition, iontophoresis was applied for 15, 30, 45 and 60min. Studies were conducted in which the formulation was either removed or left in contact with the skin following iontophoresis and then passive delivery was assessed 23h later. Tape stripping and skin extraction were performed to quantify drug levels in the stratum corneum and the underlying skin, respectively. The samples were analyzed using HPLC. The amount of drug delivered into the stratum corneum following iontophoresis was not significantly different from the amount delivered passively (p>0.05). However, drug levels in the underlying skin were significantly higher for the iontophoretic group. The amount of terbinafine delivered into the skin layers was influenced by current density and duration of current application. Leaving the drug formulation in contact with the skin during the post-iontophoretic period had a significant effect on drug levels delivered into skin layers. Iontophoresis enhanced the delivery of terbinafine hydrochloride into the skin layers and, therefore, may be used to improve the treatment of skin fungal infections. PMID- 20025951 TI - Effect of 1,4-cyclohexanediol on percutaneous absorption and penetration of azelaic acid. AB - The objective of this study is to investigate the effect of 1,4-cyclohexanediol as a retardant on the percutaneous absorption and penetration of azelaic acid. Hairless rat skin was mounted on Franz diffusion cells and treated with topical formulations containing solubilized azelaic acid with and without 1,4 cyclohexanediol. The skin was separated into stratum corneum and the deeper skin layers. The azelaic acid collected in receptor medium and each layer at the end of each time point was extracted and quantified. A significant decrease in flux across the skin suggests a penetration retardation effect of 1,4-cyclohexanediol (42.50 microg/cm(2)/h in the presence of vs. 76.25 microg/cm(2)/h in the absence of) at active loading level of 1.13 mg/cm(2). The penetration retardation effect was also observed at higher active loading level (2.82 mg/cm(2)). Furthermore, presence of 1,4-cyclohexanediol in the topical formulation did not reduce the skin and epidermal retention of azelaic acid, suggesting its potential use in the development of superior topical formation for reducing potential systematic side effect while maintaining therapeutic efficiency. PMID- 20025952 TI - Targeted sequence alteration of a chromosomal locus in mouse liver. AB - Targeted sequence alteration would be an attractive method in gene therapy and biotechnology. To achieve in vivo targeted sequence alteration, a tailed duplex DNA consisting of annealed 35mer and 794mer single-stranded DNAs was delivered by means of hydrodynamic tail vein injection into liver of transgenic mouse harboring a reporter gene (the rpsL gene) in its genome. The tailed DNA was designed for a conversion of ATC to AGC at codon 80 of the rpsL transgene. The anticipated T-->G sequence alteration was induced in the transgene in the liver with an efficiency of approximately 0.1%. These results demonstrate the significant potential of this method for applications in gene therapy and biotechnology. PMID- 20025954 TI - Neuroprotective activity of Matricaria recutita Linn against global model of ischemia in rats. AB - ETHNOPHARMACOLOGICAL RELEVANCE: Traditionally, the whole plant is used for various diseases, including neuronal disorders. AIM OF THE STUDY: To evaluate the neuroprotective effect of Matricaria recutita L. against global cerebral ischemia/reperfusion (I/R) injury-induced oxidative stress in rats. MATERIALS AND METHODS: Neuroprotective activity was carried out by global cerebral ischemia on Sprague-Dawley rats by bilateral carotid artery (BCA) occlusion for 30 min followed by 60 min reperfusion. The antioxidant enzymatic and non-enzymatic levels were estimated along with cerebral infarction area and histopathological studies. RESULTS: The Matricaria recutita L. methanolic extract showed dose dependent neuroprotective activity by significant decrease in lipid peroxidation (LPO) and increase in the superoxide dismutase (SOD), catalase (CAT), glutathione (GSH) and total thiol levels in extract treated groups as compared to ischemia/reperfusion group. Cerebral infarction area was significantly reduced in extract treated groups as compared to ischemia/reperfusion group. CONCLUSION: The methanolic extract of Matricaria recutita L. showed potent neuroprotective activity against global cerebral ischemia/reperfusion injury-induced oxidative stress in rats. PMID- 20025953 TI - Gastroprotective effect of barbatusin and 3-beta-hydroxy-3-deoxibarbatusin, quinonoid diterpenes isolated from Plectranthus grandis, in ethanol-induced gastric lesions in mice. AB - ETHNOPHARMACOLOGICAL RELEVANCE: Validate the popular use of Plectranthus grandis in gastric disorders through the active components. AIMS: Isolation of barbatusin (BB) and 3beta-hydroxy-3-deoxibarbatusin (BBOH), diterpenes from Plectranthus grandis, and evaluation of their gastroprotective effect and possible mechanisms of action. MATERIALS AND METHODS: Isolation and chemical characterization of diterpenes from Plectranthus grandis by chromatographic and spectroscopic methods and evaluation of gastroprotective action of the diterpenes through ethanol induced gastric injury in mice model. It was evaluated the effect of capsazepine, indomethacin and the role of nitric oxide and K(ATP-) channels on the gastroprotective effect of BBOH and BB. Additionally it was measured the concentrations of gastric mucus, non-proteic-sulfhydryl groups and total thiobarbituric acid-reactive substances. RESULTS: Orally administered BBOH and BB at doses of 5 and 10mg/kg, markedly reduced the gastric lesions by 59 and 96%, and 32 and 76%, respectively, with superior results as compared to N acetylcysteine (150 mg/kg, i.p.), reference compound that caused 85% lesion suppression. Although BBOH presented a higher gastroprotection than BB they act by similar mechanisms in relation to N-acetylcysteine, and prevent the depletion of gastric mucus, gastric mucosal non-proteic-sulfhydryl groups as well as the increase in thiobarbituric acid-reactive species. Moreover, the gastroprotective effect of BB was effectively blocked in mice pretreated with TRPV1 antagonist capsazepine, by the non-selective cyclooxygenase inhibitor indomethacin, or by the nitric oxide synthase inhibitor L-NAME but not by K(+)(ATP) channel inhibitor glibenclamide. In contrast, the gastroprotective effect of BBOH was blocked only by indomethacin and glibenclamide pretreatments. CONCLUSION: The protective role for BBOH and BB affording gastroprotection against gastric damage induced by ethanol indicates that these compounds contribute for the activity of Plectranthus species. The different modes of action are probably related to differences in their chemical structure. PMID- 20025955 TI - Toxicity of derivatives from semicarbazide-sensitive amine oxidase-mediated deamination of methylamine against Toxoplasma gondii after infection of differentiated 3T3-L1 cells. AB - Adipose tissue plays an active role in normal metabolic homeostasis as well as in the development of human diseases such as atherosclerosis and diabetes. We report here antimicrobial activities of the metabolites from adipocytes. Specifically, semicarbazide-sensitive amine oxidase of differentiated 3T3-L1 cells was found to utilize methylamine for producing formaldehyde and hydrogen peroxide, accounting for the inhibition of infectivity of Toxoplasma gondii and its replication in these cells. This was demonstrated by the findings that semicarbazide-sensitive amine oxidase was extremely high in differentiated 3T3-L1 cells; and that the infection of these cells by T. gondii and its intracellular replication were decreased to 33% and 37% of the control, respectively, when methylamine was provided in micromolar concentrations as the substrate to the aminoxidase. Only one of the two reaction products expected was found inhibitory against T. gondii when added to the infected pre-adipocytes of 3T3-L1. Intracellular replication of this parasite was inhibited by formaldehyde in the range of 10-100 microM and stimulated by hydrogen peroxide at 1-10 microM. The finding indicates that T. gondii may be useful as a sensitive and convenient sentinel for screening agents toxic to eukaryotic cells. PMID- 20025956 TI - Validation of an in vitro screening test for predicting the tumor promoting potential of chemicals based on gene expression. AB - Chemical carcinogenesis is a multifactorial process comprising two main stages: initiation and promotion. Tumor promoters cause the development of tumors in initiated cells and the majority of them are non-genotoxic carcinogens. The identification of tumor promoters is important for preventing cancer. We previously identified 22 specific gene markers using a global gene expression analysis of chemically induced tumor promotion and established an in vitro real time PCR screening assay for the assessment of the tumor promoting potential of chemicals in BALB/c 3T3 cells. Our in vitro tumor promoter screening test, based on these marker genes, enables earlier assessment, and is easier to conduct than classical methods. The general applicability of these markers, however, was unknown. In this study, to evaluate the performance of a set of markers, we independently validated a separate sample set, which had various structures and properties. Independent validation of the signature of 63 test chemicals showed an accuracy, sensitivity, and specificity of the assay of 96.8%, 97.0% and 96.7%, respectively. These results indicate that the tumor promoting activity assay, based on the expression of 22 marker genes, will become a valuable tool for rapid screening of potential tumor promoters. PMID- 20025957 TI - Differential sensitivity to endotoxin exposure in young and middle-age mice. AB - Aging can have a profound effect on the neurobehavioral response to immune activation; aged subjects are predisposed to greater deficits in performance and cognitive function in conjunction with an exaggerated neuroinflammatory response. While increased reactivity to an immune insult has been well characterized in aged subjects, the alterations that may exist by middle-age have not been thoroughly investigated. The present study compared the reactions of young (4 month) and middle-age (12-month) male BALB/c mice to an acute or repeated lipopolysaccharide (LPS) challenge(s). The data suggest that in some respects middle-aged mice are more sensitive to endotoxin exposure, as they show enhanced weight loss, splenic cytokine levels, and c-fos expression in the brain following acute LPS administration compared to younger mice. However, acute LPS exposure led to comparable decreases in locomotor activity in young and middle-aged mice. Following repeated LPS administration both age groups showed diminished behavioral and neural reactions to the final LPS challenge, indicating tolerance development. However, the immune system of the middle-aged mice was still mildly responsive to the final LPS exposure, as splenic levels of IL-1beta were significantly elevated. Collectively, the data suggest that middle-age subjects are more sensitive to an immune insult. PMID- 20025958 TI - Involvement of phosphoinositide 3-kinase gamma in the neuro-inflammatory response and cognitive impairments induced by beta-amyloid 1-40 peptide in mice. AB - Alzheimer disease (AD) is the most common form of dementia in the elderly, and the neuro-pathological hallmarks of AD include neurofibrillary tangles (NFT), and deposition of beta-amyloid (Abeta) in extracellular plaques. In addition, chronic inflammation due to recruitment of activated glial cells to amyloid plaques are an invariant component in AD, and several studies have reported that the use of non-steroidal anti-inflammatory drugs (NSAIDs) may provide a measure of protection against AD. In this report we have investigated whether phosphoinositide 3-kinase gamma (PI3Kgamma), which is important in inflammatory cell migration, plays a critical role in the neuro-inflammation, synaptic dysfunction, and cognitive deficits induced by intracerebroventricular injection of Abeta(1-40) in mice. We found that the selective inhibitor of PI3Kgamma, AS605240, was able to attenuate the Abeta(1-40)-induced accumulation of activated astrocytes and microglia in the hippocampus, and decrease immuno-staining for p Akt and cyclooxygenase-2 (COX-2). Interestingly, Abeta(1-40) activated macrophages treated with AS605240 or another PI3Kgamma inhibitor, AS252424, displayed impaired chemotaxis in vitro, but their expression of tumor necrosis factor-alpha (TNF-alpha) and interleukin-1beta (IL-1beta) was unaffected. Finally, AS605240 prevented Abeta(1-40)-induced cognitive deficits and synaptic dysfunction, but failed to modify scopolamine-induced amnesia. Our data suggests that inhibition of PI3Kgamma may represent a novel therapeutic target for treating AD patients. PMID- 20025959 TI - Embryonic toxicokinetic and dynamic differences underlying strain sensitivity to cadmium during neurulation. AB - Differences in sensitivity are observed between mouse strains, C57 (sensitive) and SWV (resistant) when exposed to cadmium (Cd) during the neurulation period. In this study, we investigated the toxicokinetics of Cd in relation with toxicodynamic responses to identify factors affecting differential Cd-sensitivity in C57 and SWV. Using a level of exposure which induced developmental toxicity and differential effects between strains, we assessed maternal and embryonic Cd uptake and evaluated biomarkers of response previously linked with Cd exposure, specifically metal ion regulators (Mt1, Mt2, DMT1) and markers of cell cycle arrest/apoptosis induction (p53, Cdkn1a, c-Casp3). Greater Cd uptake was observed in C57 embryos compared to SWV and these observations of differential uptake were associated with increased alterations in expression of biomarkers of metal response (e.g. c-Casp3) and strain sensitivity. Using sensitive and resistant mouse strains, we have identified toxicokinetic and dynamic differences which underlie observed differences in Cd embryonic sensitivity and response. PMID- 20025960 TI - Rapid diagnosis of spinal muscular atrophy using tetra-primer ARMS PCR assay: simultaneous detection of SMN1 and SMN2 deletion. AB - Spinal muscular atrophy (SMA), the leading genetic cause of death in childhood, is an autosomal recessive neuromuscular disorder characterized by progressive muscle weakness, associated with deletions of the survival motor neuron 1 (SMN1) gene. Approximately 94% of SMA patients carry homologous deletions of SMN1 exon(s) 7 (and 8). Because of the high incidence and severity of the disease, precise detection and quantification of SMN1 and SMN2 gene copy numbers is essential for diagnosis and genetic counseling. We have developed a reliable single-tube tetra-primer PCR assay to simultaneously detect both the SMN1 and SMN2 exon 7 deletion using the advantage of C/T difference at nucleotide position of 840 in exon 7. The assay has been optimized and tested in 48 healthy controls, 20 known patients with SMA, 12 carriers (one SMN1 copy), and 8 amniotic fluids suspected of having SMA for whom we had determined the SMN1/SMN2 deletion by an additional PCR-RFLP method. We have observed complete concordance between methods. Our tetra-primer PCR assay is sensitive, low-cost, and easy to use method for simultaneous detection of both SMN1 and SMN2 deletion, which could be used even in "low-tech" laboratories. PMID- 20025961 TI - Recombinant OMP28 antigen-based indirect ELISA for serodiagnosis of bovine brucellosis. AB - Brucellosis is a zoonosis of both public health and economic importance in many developing countries including India. Early detection and segregation of the infected animals are important in order to control the disease. Serodiagnostic tests for brucellosis is mainly based on detection of antibodies developed against lipopolysaccharide (LPS) component of cell. In this study we evaluated a protein antigen, 28 kDa outer membrane protein (OMP28), of Brucella melitensis as an alternative to LPS. Recombinant OMP28 was produced in Escherichia coli system. The efficacy of purified OMP28 was studied in an indirect enzyme-linked immunosorbent assay (ELISA) for diagnosis of brucellosis in field sera collected from different regions of country. Using known negative and known positive serum samples it was found that OMP28 is immunoreactive to Brucella infected cattle, sheep, goat and dog sera. Three hundred and eighty two cattle sera were screened by OMP28 antigen-based ELISA and the results were compared to rose Bengal plate agglutination Test (RBPT). Recombinant OMP28 antigen-based ELISA has shown sensitivity of 88.7%, specificity of 93.8% and accuracy of 92.9%. It was concluded that recombinant B. melitensis OMP28 could be used as a protein antigen for diagnosis of brucellosis in domestic animals. PMID- 20025962 TI - Changes in bladder innervation in a mouse model of Alzheimer's disease. AB - AIM: The aims of this study were to compare the structure of bladders from a transgenic mouse model of Alzheimer's disease with age matched control animals and to explore the idea that any structural differences might be related to functional bladder changes associated with the condition. MATERIALS AND METHODS: Two groups of mice were used. Transgenic animals in which the murine Amyloid Precursor Protein (APP) gene has been partly replaced by the human APP including both the Swedish and London mutations and that overexpress a mutant of the human Presenilin 1 gene (PS1M146L) driven by the PDGF promoter. The transgenic mice (App(SL)/PS1(M146L)) aged 24+/-3 months were used. The second group was an age matched control group of C57 black mice. The bladders from each group were isolated, fixed in 4% paraformaldehyde and prepared for immunohistochemistry. Antibodies to the vesicular acetylcholine transporter (VAChT) and neuronal nitric oxide synthase (nNOS) were used to identify neural structures. RESULTS: Cholinergic nerves (VAChT(+)) were observed in the inner and outer muscle bundles of App(SL)/PS1(M146L) and control mice. No major differences were noted in the distribution of these fibres. In contrast, there was a distinct difference in the innervation of the sub-urothelial layer. In App1(SL)/PS1(M146L) mice there were numerous VAChT and nNOS positive fibres in sharp contrast to the paucity of similar nerves in control animals. VAChT and nNOS did not appear to co-localise in the same nerve fibres within the lamina propria. Pairs of nerve fibres, nNOS(+) and VAChT(+), were observed to be intertwined and run in close proximity. A particularly unusual feature of the App(SL)/PS1(M146L) mouse bladder was the presence of neurones within the bladder wall. These nerve cell bodies were seen in all App(SL)/PS1(M146L) mouse bladders. The neurones could be found singly or in small ganglion like groups of cells and were located in all layers of the bladder wall (sub-urothelium, in the lamina propria adjacent to the inner muscle and within the inner muscle and outer muscle layers). No nerve cells or small ganglia were noted in any of the control bladders studied. CONCLUSIONS: There are structural differences in the bladders of App(SL)/PS1(M146L) mice compared to control animals. These differences are associated with sub-urothelial nerves which, because of their location, are likely to be sensory fibres. This may lead to a changed sensory processing from the App(SL)/PS1(M146L) bladders. The physiological role of the intra-mural neurones and ganglia is not known. It is speculated that they may be associated with peripheral motor/sensory mechanisms linked to the generation and modulation of sensation. PMID- 20025963 TI - Chromatin repair after oxidative stress: role of PARP-mediated proteasome activation. AB - Oxidative stress is an inevitable process in the nucleus, especially in antitumor chemotherapy, and adaptation by defense mechanisms seems to be one element in the development of long-term resistance to many chemotherapeutic drugs. In this study, a potential chromatin repair mechanism during oxidative stress was investigated in HT22 cells. The 20S proteasome has been shown to be largely responsible for the degradation of oxidatively modified histone proteins in the nucleus. Poly(ADP-ribosyl)ation reactions also play an important role in DNA repair as a consequence of oxidative damage and single-strand breaks. Such a reaction may occur also with the 20S proteasome--with a known increase in enzymatic activity--and also with histones--reducing their proteolytic susceptibility as shown for the first time here. After hydrogen peroxide treatment of HT22 cells, degradation of the model peptide substrate suc-LLVY-MCA and degradation of oxidized histones by nuclear proteasome increased. During the removal of protein carbonyls, single-strand breaks and 8-hydroxy-2' deoxyguanosine, proteasome, and poly(ADP-ribose) polymerase-1 enzymes were shown to play tightly interacting roles. Our results following the repair of oxidative damage show the proteolytic activation of proteasome concerning poly(ADP ribosyl)ation together with a decline in poly(ADP-ribosyl)ation of oxidized histones, leading to a selective recognition of oxidatively modified histones. PMID- 20025964 TI - Association of urinary 15-F2t-isoprostane level with oxygen desaturation and carotid intima-media thickness in nonobese sleep apnea patients. AB - Obstructive sleep apnea (OSA) is characterized by recurrent apnea during sleep that may unbalance oxidative stress, increasing atherosclerosis. Among oxidative stress markers, 15-F(2t)-isoprostane is considered one of the most sensitive and specific metabolites of lipid peroxidation. To explore the relationship between urinary 15-F(2t)-isoprostane with sleep apnea severity and carotid modifications in nonobese OSA patients, 31 nonobese sleep apnea patients were studied, along with 10 lean subjects without OSA. Patients were assessed by polysomnography, blood pressure measurement, and ultrasonography to determine the carotid intima media thickness (IMT). Urinary 15-F(2t)-isoprostanes were measured by liquid chromatography-tandem mass spectrometry. Urinary 15-F(2t)-isoprostane concentrations were increased in severe OSA patients compared to control subjects (20.2+/-7.3 vs 12.3+/-2.8 ng/mmol creatinine; P=0.020). Mean carotid IMT was correlated with 15-F(2t)-isoprostane (r=0.532; P<0.001) and with the apnea hypopnea index (r=0.345; P=0.029). 15-F(2t)-Isoprostane level was related to the night time spent at SaO(2)<90% (r=0.478; P=0.002), the apnea-hypopnea index (r=0.465; P=0.003), and the mean nocturnal SaO(2) (r=-0.424; P=0.007). These results showed a relationship between lipid peroxidation, carotid intima-media thickness, and intermittent hypoxia in nonobese OSA patients, thus reinforcing the hypothesis that oxidative stress could be involved in the early atherosclerotic process. PMID- 20025965 TI - Effects of age and calorie restriction on tryptophan nitration, protein content, and activity of succinyl-CoA:3-ketoacid CoA transferase in rat kidney mitochondria. AB - This study examined the protein targets of nitration and the consequent impact on protein function in rat kidney mitochondria at 4, 13, 19, and 24 months of age. Succinyl-CoA transferase (SCOT), a rate-limiting enzyme in the degradation of ketone bodies, was the most intensely reactive protein against anti-3 nitrotyrosine antibody in rat kidney mitochondria. However, subsequent mass spectrometric and amino acid analyses of purified SCOT indicated that tryptophan 372, rather than a tyrosine residue, was the actual site of simultaneous additions of nitro and hydroxy groups. This finding suggests that identification of nitrated tyrosine residues based solely on reactivity with anti-3 nitrotyrosine antibody can be potentially misleading. Between 4 and 24 months of age, the amounts of SCOT protein and catalytic activity, expressed per milligram of mitochondrial proteins, decreased by 55 and 45%, respectively. SCOT, and particularly its nitrated carboxy-terminal region, was relatively more susceptible to in vitro proteolysis than other randomly selected kidney mitochondrial proteins. The age-related decreases in SCOT protein amount and catalytic activity were prevented by a relatively long-term 40% reduction in the amount of food intake. Loss of SCOT protein in the aged rats may attenuate the capacity of kidney mitochondria to utilize ketone bodies for energy production. PMID- 20025966 TI - Physiologically based mechanistic modelling to predict complex drug-drug interactions involving simultaneous competitive and time-dependent enzyme inhibition by parent compound and its metabolite in both liver and gut - the effect of diltiazem on the time-course of exposure to triazolam. AB - AIM: To predict the magnitude of metabolic drug-drug interaction (mDDI) between triazolam and diltiazem and its primary metabolite N-desmethyldiltiazem (MA). METHODS: Relevant in vitro metabolic and inhibitory data were incorporated into a mechanistic physiologically based pharmacokinetic model within Simcyp (Version 9.1) to simulate the time-course of changes in active CYP3A4 content in gut and liver and plasma concentrations of diltiazem, MA and triazolam in a virtual population with characteristics related to in vivo studies. RESULTS: The predicted median increases in AUC(0,infinity) of triazolam, which ranged from 3.9 to 9.5 for 20 simulated trials (median 5.9), were within 1.5-fold of the observed median value (4.4) in 14 of the trials. Considering the effects of diltiazem only and not those of MA, and ignoring auto-inhibition of MA metabolism and inhibition of its metabolism by diltiazem, resulted in lower increases in triazolam exposure (AUC ratios of 1.5-2.0 (median 1.7) and 2.7-5.3 (median 3.4), respectively). CONCLUSION: Prediction of mDDIs involving diltiazem requires consideration of both competitive and time-dependent inhibition in gut and liver by both diltiazem and MA, as well as the complex interplay between the two moieties with respect to mutual inhibition of parent compound and its metabolite. PMID- 20025967 TI - Mechanism of hydrolysis of a novel indolocarbazole topoisomerase I inhibitor. AB - The degradation kinetics and reaction product profile of the antitumor agent 1 in aqueous solution was studied. Hydrolysis of the pendant imide ring of 1 is the primary mode of thermal degradation in aqueous solution, and the pH rate profile of 1 has a V-shape indicating that hydrolysis of the imide ring can be catalyzed by either acid or base. Hydrolysis of 1 to the anhydride derivative 3 or the dicarboxylic acid derivative 4 is stepwise and the intermediates 2a and 2b formed by initial hydrolytic attack have been observed under alkaline conditions. An overall mechanism for the hydrolysis of 1 in aqueous solution has been proposed. Extrapolating Arrhenius behavior to the hydrolysis reaction of 1 in aqueous solution maintained at a pH value of 4 suggests an aqueous buffered formulation has sufficient thermal stability to be considered a robust room temperature drug product. PMID- 20025968 TI - Cytotoxicity study of ordered mesoporous silica MCM-41 and SBA-15 microparticles on Caco-2 cells. AB - Cytotoxicity of ordered mesoporous silica MCM-41 and SBA-15 microparticles (fractions between 1 and 160 microm) was determined in vitro on undifferentiated human colon carcinoma (Caco-2) cell line, considering the feasibility of using these silica-based materials in oral drug formulations. The cellular endpoints employed for assessing the effects of the MCM-41 and SBA-15 microparticles on Caco-2 were: (1) cell membrane integrity by monitoring live-cell protease activity (AFC) and by employing the flow cytometry method; (2) metabolic activity by monitoring total ATP content via luminescence assay; (3) activity of apoptotic effectors by caspase-3/7 activity assay. The generation of reactive oxygen species (ROS) was also followed, specifically the hydrogen peroxide (H(2)O(2)) and the superoxide radical (O(2)(-)). MCM-41 and SBA-15 microparticles caused cytotoxic effects on the Caco-2 cells, at most tested concentrations (0.2-14 mg/ml) and incubation times (3 and 24h). The effects on the cells included weakened cell membrane integrity, diminished cell metabolism and increased apoptotic signalling. The root cause for the cytotoxicity was heightened production of reactive oxygen species (ROS), especially the formation of the superoxide radical O(2)(-) already after 3h incubation with threshold dose 1mg/ml, apparently overwhelming the antioxidant defences and causing mitochondrial dysfunction, hence increasing the apoptotic signalling. PMID- 20025969 TI - Isolation and characterization of three estrogen receptor transcripts in Oreochromis mossambicus (Peters). AB - Exposure of aquatic organisms to 17beta-estradiol (E(2)) induces a variety of estrogen-responsive genes, including vitellogenin (vtg)-the precursor protein of egg yolk in oviparous animals and to date the single most used gene product in screening for estrogenic endocrine disruption. Transcription regulation of vtg by E(2) is dependent on binding of the ligand (E(2)) to a specific nuclear receptor (estrogen receptor, ESR) which in turn binds to an estrogen responsive element (ERE) in the promoter of vtg. Since a local tilapiine, Oreochromis mossambicus (Peters), is targeted as a model for estrogenic endocrine disruption in Southern Africa, a platform of knowledge is necessary for the ontogenic and tissue specific behavior of ESR in this species before vtg levels can be interpreted in relation to such endocrine disruption. Therefore, three ESR cDNA sequences (ESR1, ESR2a and ESR2b) in O. mossambicus were isolated and QPCR protocols were developed to ascertain their quantitative transcript levels in adult brain, gonadal and hepatic tissues. ESR1 transcript levels were highest in female liver tissue compared to males and other tissues, whereas the levels for ESR2a and b were not statistically significantly different between male and female tissues. Quantitative gene levels during development demonstrated a sharp increase in ESR1 during the stage of gonad differentiation (50-60 days post-fertilization) in this species. Finally, an induction experiment in adult male liver tissue confirms the upregulation of ESR1 by E(2). PMID- 20025970 TI - Beta-amyloid controls altered Reelin expression and processing in Alzheimer's disease. AB - Reelin is a glycoprotein that modulates synaptic function and plasticity in the mature brain, thereby favouring memory formation. We recently reported altered cerebral Reelin expression in Alzheimer's disease (AD). Here we demonstrate pronounced Reelin changes at protein and mRNA levels in the frontal cortex in adult Down's syndrome (DS), where the extra copy of chromosome 21 leads to overexpression of beta-amyloid. In cortical extracts of fetal DS samples we detected increased levels of the full-length Reelin and the 310-kDa fragment. Overexpression of mutant human amyloid precursor protein also led to an increase in levels of Reelin fragments in Tg2576 transgenic mice for human beta-amyloid. Finally, in vitro Abeta42 treatment of SH-SY5Y neuroblastoma cells led to increased Reelin levels. An altered pattern of Reelin glycosylation was detected in extracts from the frontal cortex of AD patients and in Abeta42-treated SH-SY5Y cells, supporting the notion that beta-amyloid triggers altered Reelin processing. These results provide evidence that Reelin expression and processing is altered in several amyloid conditions. PMID- 20025971 TI - VEGF up-regulation by G93A superoxide dismutase and the role of malate-aspartate shuttle inhibition. AB - A gain of interaction of the amyotrophic lateral sclerosis (ALS)-linked G93A superoxide dismutase-1 (G93A-hSOD1) with cytosolic malate dehydrogenase (cytMDH), a key enzyme in the malate-aspartate shuttle, diverts neurons towards anaerobic metabolism. Changes in vascular endothelial growth factor (VEGF) are reported in ALS and hypoxia. Here we report that expression of G93A-hSOD1 fused with green fluorescent protein in NSC-34 cells enhanced VEGF expression and levels of VEGF and its upstream regulator hypoxia-inducible factor (HIF-1alpha). G93A-hSOD1 expressing cells were unable to further up-regulated VEGF in response to Co(2+) and H(2)O(2). Amino-oxyacetate that inhibits the malate-aspartate shuttle caused a similar increase in VEGF mRNA and impaired response to H(2)O(2) in WT-hSOD1 expressing cells. Interruption of the G93A-hSOD1/cytMDH interaction reduced VEGF expression in G93A-hSOD1 expressing cells and restored their ability to up regulate VEGF in response to Co(2+) and H(2)O(2). These results demonstrate that the ALS-linked G93A hSOD1 mutation impairs VEGF regulation compatible with the inhibition of neuronal malate-aspartate shuttle. PMID- 20025972 TI - Why the need for qPCR publication guidelines?--The case for MIQE. AB - The polymerase chain reaction (PCR) has matured from a labour- and time intensive, low throughput qualitative gel-based technique to an easily automated, rapid, high throughput quantitative technology. Real-time quantitative PCR (qPCR) has become the benchmark technology for the detection and quantification of nucleic acids in a research, diagnostic, forensic and biotechnology setting. However, ill-assorted pre-assay conditions, poor assay design and inappropriate data analysis methodologies have resulted in the recurrent publication of data that are at best inconsistent and at worst irrelevant and even misleading. Furthermore, there is a lamentable lack of transparency of reporting, with the "Materials and Methods" sections of many publications, especially those with high impact factors, not fit for the purpose of evaluating the quality of any reported qPCR data. This poses a challenge to the integrity of the scientific literature, with serious consequences not just for basic research, but potentially calamitous implications for drug development and disease monitoring. These issues are being addressed by a set of guidelines that propose a minimum standard for the provision of information for qPCR experiments ("MIQE"). MIQE aims to restructure to-day's free-for-all qPCR methods into a more consistent format that will encourage detailed auditing of experimental detail, data analysis and reporting principles. General implementation of these guidelines is an important requisite for the maturing of qPCR into a robust, accurate and reliable nucleic acid quantification technology. PMID- 20025973 TI - Inhibition of Toll-like receptors TLR4 and 7 signaling pathways by SIGIRR: a computational approach. AB - Toll-like receptors (TLRs) belong to the Toll-like receptor/interleukin-1 receptor (TLR/IL-1R) superfamily which is defined by a common cytoplasmic Toll/interleukin-1 receptor (TIR) domain. TLRs recognize pathogen-associated molecular patterns and initiate an intracellular kinase cascade to trigger an immediate defensive response. SIGIRR (single immunoglobulin interleukin-1 receptor-related molecule), another member of the TLR/IL-1R superfamily, acts as a negative regulator of MyD88-dependent TLR signaling. It attenuates the recruitment of MyD88 adaptors to the receptors with its intracellular TIR domain. Thus, SIGIRR is a highly important molecule for the therapy of autoimmune diseases caused by TLRs. So far, the structural mechanism of interactions between SIGIRR, TLRs and adaptor molecules is unclear. To develop a working hypothesis for this interaction, we constructed three-dimensional models for the TIR domains of TLR4, TLR7, MyD88 and SIGIRR based on computational modeling. Through protein protein docking analysis, we developed models of essential complexes involved in the TLR4 and 7 signaling and the SIGIRR inhibiting processes. We suggest that SIGIRR may exert its inhibitory effect through blocking the molecular interface of TLR4, TLR7 and the MyD88 adaptor mainly via its BB-loop region. PMID- 20025974 TI - A novel approach to the structural analysis of partially decorated actin based filaments. AB - We describe a novel set of single particle based procedures for the structural analysis of electron microscope images of muscle thin filaments and other partially decorated actin based filaments. The thin filament comprises actin and the regulatory proteins tropomyosin and troponin in a 7:1:1M ratio. Prior to our work, structure analysis from electron microscope images of the thin filament has largely involved either helical averaging defined by the underlying actin helix or the use of single particle analysis but using a starting model as a reference structure. Our single particle based approach yields an accurate structure for the complete thin filament by avoiding the loss of information from troponin and tropomyosin associated with helical averaging and also removing the potential reference bias associated with the use of a starting model. The approach is more widely applicable to sub-stoichiometric complexes of F-actin and actin-binding proteins. PMID- 20025975 TI - Cryo-electron tomography of microtubule-kinesin motor complexes. AB - Microtubules complexed with molecular motors of the kinesin family or non-motor microtubule associated proteins (MAPs) such as tau or EB1 have been the subject of cryo-electron microcopy based 3-D studies for several years. Most of these studies that targeted complexes with intact microtubules have been carried out by helical 3-D reconstruction, while few were analyzed by single particle approaches or from 2-D crystalline arrays. Helical reconstruction of microtubule-MAP or motor complexes has been extremely successful but by definition, all helical 3-D reconstruction attempts require perfectly helical assemblies, which presents a serious limitation and confines the attempts to 15- or 16-protofilament microtubules, microtubule configurations that are very rare in nature. The rise of cryo-electron tomography within the last few years has now opened a new avenue towards solving 3-D structures of microtubule-MAP complexes that do not form helical assemblies, most importantly for the subject here, all microtubules that exhibit a lattice seam. In addition, not all motor domains or MAPs decorate the microtubule surface regularly enough to match the underlying microtubule lattice, or they adopt conformations that deviate from helical symmetry. Here we demonstrate the power and limitation of cryo-electron tomography using two kinesin motor domains, the monomeric Eg5 motor domain, and the heterodimeric Kar3Vik1 motor. We show here that tomography does not exclude the possibility of post-tomographic averaging when identical sub-volumes can be extracted from tomograms and in both cases we were able to reconstruct 3-D maps of conformations that are not possible to obtain using helical or other averaging-based methods. PMID- 20025976 TI - Cloning of common carp SOCS-3 gene and its expression during embryogenesis, GH transgene and viral infection. AB - As a member of a newly discovered protein family, the suppressor of cytokine signalling 3 (SOCS-3) has been shown to regulate the responses of many immune cytokines in a negative auto-regulatory manner. The full-length cDNA of common carp SOCS-3 was 1603 bp and contained a 630 bp open reading frame (ORF) coding for a protein of 209 amino acids. Carp SOCS-3 molecule was well conserved especially in the SRC homology 2 (SH2) and the SOCS box. The kinase inhibitory region (KIR) and ESS domains, upstream of the SH2 domain, were conserved in carp SOCS-3, except for a specific insertion (PHRYK) in the KIR domain at the N terminal region. Three conserved cysteine (Cys-102, 124 and 193) residues, and one additional cysteine (Cys-168) residue, were also found in carp SOCS-3. The 2015 bp genomic DNA of carp SOCS-3 contained two exons and one intron. Phylogenetic analysis showed that carp SOCS-3 sequence grouped with other known fish SOCS-3 sequences with zebrafish SOCS-3 as the closest neighbour. RT-PCR analysis showed that carp SOCS-3 was initially expressed at 4 h pf (post fertilization) and gradually increased up to 4 w pf during embryogenesis. By RT qPCR analysis, carp SOCS-3 gene was predominantly detected in gill, head kidney, thymus and skin, followed by spleen and peripheral blood, lower expression level was detected in kidney, intestine, liver and muscle; the SOCS-3 transcript was significantly increased in thymus, head kidney, spleen and intestine of GH (growth hormone)-transgenic carp; after SVCV (spring viraemia of carp virus) infection, the carp SOCS-3 transcript was significantly up-regulated in gill, intestine, thymus, spleen, head kidney and kidney tissues in a time-dependent manner. These results suggest that teleost SOCS-3 may play an active role in the modulation of viral-induced innate immune response and in preventing the overaction of some cytokines with viral stimulation. PMID- 20025977 TI - Immune response and expression analysis of cathepsin K in goldfish during Aeromonas hydrophila infection. AB - The innate immunity and expression profiles of cathepsins D were determined in the goldfish (Carassius auratus) tissues after challenge with a fish pathogen Aeromonas hydrophila. The innate immunity of reactive oxygen species (ROS) and reactive nitrogen species (RNS) were determined by peripheral blood leucocytes. Blood and tissue samples of the muscle, gills, liver, kidney, heart, spleen, and intestine were sampled at 1, 3, 6 and 12 h post-infection for cathepsin D expression by semi-quantitative RT-PCR. The ROS and RNS production did not significantly increase at 1 h post-challenged goldfish. However, the ROS and RNS production was significantly increased after 3 h post-challenged fish compared to the control. The cathepsin D expression was found very low in muscle and kidney of the control fish, other tissues was not found the expression. A similar pattern was found in goldfish at 1 h post-challenge with A. hydrophila. However, at 3 h post-challenge goldfish, the cathepsin D expression was high only in the heart. At 6 h post-challenge goldfish, the cathepsin D expression was seen high all the tissues, except in the spleen. However, the expression was decreased at 12 h post-infection samples. This result was suggested that the goldfish infected with A. hydrophila decreased the innate immunity level in peripheral blood and expressed the cathepsin D in tissues. PMID- 20025978 TI - An algebra for the analysis of object encoding. AB - The encoding of the objects from the world around us is one of the major topics of cognitive psychology, yet the principles of object coding in the human brain remain unresolved. Beyond referring to the particular features commonly associated with objects, our ability to categorize and discuss objects in detailed linguistic propositions implies that we have access to generic concepts of each object category with well-specified boundaries between them. Consideration of the nature of generic object concepts reveals that they must have the structure of a probabilistic list array specifying the Bayesian prior on all possible features that the object can possess, together with mutual covariance matrices among the features. Generic object concepts must also be largely context independent for propositions to have communicable meaning. Although, there is good evidence for local feature processing in the occipital lobe and specific responses for a few basic object categories in the posterior temporal lobe, the encoding of the generic object concepts remains obscure. We analyze the conceptual underpinnings of the study of object encoding, draw some necessary clarifications in relation to its modality-specific and amodal aspects, and propose an analytic algebra with specific reference to functional Magnetic Resonance Imaging approaches to the issue of how generic (amodal) object concepts are encoded in the human brain. PMID- 20025979 TI - Power spectrum scale invariance quantifies limbic dysregulation in trait anxious adults using fMRI: adapting methods optimized for characterizing autonomic dysregulation to neural dynamic time series. AB - In a well-regulated control system, excitatory and inhibitory components work closely together with minimum lag; in response to inputs of finite duration, outputs should show rapid rise and, following the input's termination, immediate return to baseline. The efficiency of this response can be quantified using the power spectrum density's scaling parameter beta, a measure of self-similarity, applied to the first derivative of the raw signal. In this study, we adapted power spectrum density methods, previously used to quantify autonomic dysregulation (heart rate variability), to neural time series obtained via functional MRI. The negative feedback loop we investigated was the limbic system, using affect-valent faces as stimuli. We hypothesized that trait anxiety would be related to efficiency of regulation of limbic responses, as quantified by power law scaling of fMRI time series. Our results supported this hypothesis, showing moderate to strong correlations of trait anxiety and beta (r=0.45-0.54) for the amygdala, orbitofrontal cortex, hippocampus, superior temporal gyrus, posterior insula, and anterior cingulate. Strong anticorrelations were also found between the amygdala's beta and wake heart rate variability (r=-0.61), suggesting a robust relationship between dysregulated limbic outputs and their autonomic consequences. PMID- 20025981 TI - Learning to appreciate others: neural development of cognitive perspective taking. AB - Neuroimaging studies have thoroughly investigated brain regions that are recruited when we put ourselves in another person's shoes. Taking a third-person perspective (3PP) as opposed to a first-person perspective (1PP) has been associated with brain activation in the inferior parietal cortex, the medial posterior cortex and the prefrontal cortex. Here we investigate for the first time the development of the neural network that yields cognitive perspective taking. Twelve adults (aged 25-32 years) and twelve school-aged children (aged 8 10 years) were investigated using functional magnetic resonance imaging (fMRI). Behaviorally, we found a decrease of reaction time differences between 3PP and 1PP with age indicating that adults were more efficient in processing a 3PP. Despite the reaction time differences both groups were equally accurate in their judgments. Brain imaging data indicated neural activity in the left inferior parietal cortex and precuneus for adults during 3PP as compared with 1PP judgments. Children additionally showed enhanced activity in the dorsolateral prefrontal cortex and the right inferior perietal cortex. We found a significant interaction between groups and brain activation in the right dorsolateral prefrontal cortex and in the right inferior parietal cortex. These results suggest that the development of the ability to reason about another person's mind accompanies a shift in activity from frontal to posterior brain regions and from bilateral to unilateral left inferior parietal cortex. PMID- 20025980 TI - Evaluating functional localizers: the case of the FFA. AB - Functional localizers are routinely used in neuroimaging studies to test hypotheses about the function of specific brain areas. The specific tasks and stimuli used to localize particular regions vary widely from study to study even when the same cortical region is targeted. Thus, it is important to ask whether task and stimulus changes lead to differences in localization or whether localization procedures are largely immune to differences in tasks and contrasting stimuli. We present two experiments and a literature review that explore whether face localizer tasks yield differential localization in the fusiform gyrus as a function of task and contrasting stimuli. We tested standard localization tasks-passive viewing, 1-back, and 2-back memory tests--and did not find differences in localization based on task. We did, however, find differences in the extent, strength and patterns/reliabilities of the activation in the fusiform gyrus based on comparison stimuli (faces vs. houses compared to faces vs. scrambled stimuli). PMID- 20025982 TI - Learning second language vocabulary: neural dissociation of situation-based learning and text-based learning. AB - Second language (L2) acquisition necessitates learning and retrieving new words in different modes. In this study, we attempted to investigate the cortical representation of an L2 vocabulary acquired in different learning modes and in cross-modal transfer between learning and retrieval. Healthy participants learned new L2 words either by written translations (text-based learning) or in real-life situations (situation-based learning). Brain activity was then measured during subsequent retrieval of these words. The right supramarginal gyrus and left middle frontal gyrus were involved in situation-based learning and text-based learning, respectively, whereas the left inferior frontal gyrus was activated when learners used L2 knowledge in a mode different from the learning mode. Our findings indicate that the brain regions that mediate L2 memory differ according to how L2 words are learned and used. PMID- 20025983 TI - Molecular systematics of the bubblegum coral genera (Paragorgiidae, Octocorallia) and description of a new deep-sea species. AB - Bubblegum octocorals (Paragorgia and Sibogagorgia) play an important ecological role in many deep-sea ecosystems. However, these organisms are currently threatened by destructive fishing methods such as bottom trawling. Taxonomic knowledge of conservation targets is necessary for the creation and implementation of efficient conservation strategies. However, for most deep-sea coral groups this knowledge remains incomplete. For instance, despite its similarities with Paragorgia, Sibogagorgia is particular in lacking polyp sclerites, which are present in groups like Paragorgia and the Coralliidae. Although two kinds of sclerites are very similar between Paragorgia and Sibogagorgia, other characters challenge the monophyly of these genera. Here we help to clarify the taxonomy and evolutionary relationships of the bubblegum octocorals and related taxa by examining molecular data. We employed nucleotide sequences of mitochondrial (ND6, ND6-ND3 intergenic spacer, ND3, ND2, COI, msh1 and 16S) and nuclear (28S and ITS2) genomic regions from several taxa to infer molecular phylogenetics and to examine the correspondence of morphological features with the underlying genetic information. Our data strongly supported the monophyly of the genus Paragorgia, the family Coralliidae (precious corals), and a group of undescribed specimens resembling Sibogagorgia. Further morphological observations were congruent regarding the uniqueness of the undescribed specimens, here defined as a new species, Sibogagorgia cauliflora sp. nov., which occurs in both sides of the North American landmass at depths below 1700 m. This new species resembles S. dennisgordoni with branching in one plane but has fairly different radiate sclerites and significantly divergent DNA sequences. The existence of several diagnostic characters of Sibogagorgia in S. cauliflora indicates that they indeed belong to this genus. It is however remarkable that a small number of medullar canals are also found in this species; medullar canals have been considered as the main diagnostic character of Paragorgia. Thus, the evidence generated here indicates that the presence or absence of these canals per se is not a conclusively diagnostic character for either genus. The lack of internal-node resolution in the inferred phylogenetic hypotheses of these genera does not allow us to propose a clear scenario regarding the evolution of these traits. PMID- 20025984 TI - The role of anaerobic bacteria in bacteremia. AB - Anaerobic bacteria remain an important cause of bloodstream infections and account for 1-17% of positive blood cultures. This review summarizes the epidemiology, microbiology, predisposing conditions, and treatment of anaerobic bacteremia (AB) in newborns, children, adults and in patients undergoing dental procedures. The majority of AB are due to Gram-negative bacilli, mostly Bacteroides fragilis group. The other species causing AB include Peptostreptococcus, Clostridium spp., and Fusobacterium spp. Many of these infections are polymicrobial. AB in newborns is associated with prolonged labor, premature rupture of membranes, maternal amnionitis, prematurity, fetal distress, and respiratory difficulty. The predisposing conditions in children include: chronic debilitating disorders such as malignant neoplasm, hematologic abnormalities, immunodeficiencies, chronic renal insufficiency, or decubitus ulcers and carried a poor prognosis. Predisposing factors to AB in adults include malignant neoplasms, hematologic disorders, transplantation of organs, recent gastrointestinal or obstetric gynecologic surgery, intestinal obstruction, diabetes mellitus, post-splenectomy, use of cytotoxic agents or corticosteroids, and an undrained abscess. Early recognition and appropriate treatment of these infections are of great clinical importance. PMID- 20025986 TI - Cytomegalovirus glycoprotein B genotype in hematopoietic stem cell transplant patients from China. AB - Cytomegalovirus (CMV) can be classified into 4 subgroups based on genotype variation of the glycoprotein B (gB) encoded by UL55 gene. Little is known about the CMV gB genotype distribution and its clinical implication in patients who receive hematopoietic stem cell transplant (HSCT) in China. In this study, which comprises 101 HSCT patients with CMV infection, we have found that 36 patients (35.64%) were infected with CMV genotype gB1, 3 patients (2.97%) with gB2, 39 patients (38.61%) with gB3, 1 patient (0.99%) with gB4, and 17 patients (16.83%) were infected with mixed CMV genotypes. We also found that CMV gB3 was associated with a high risk of CMV pneumonitis; nevertheless, there were no significant differences among patients with gB genotypes with respect to the other CMV diseases; no significant differences between patients with gB genotypes with respect to type II-IV acute graft-verses-host disease (aGVHD) and chronic GVHD (cGVHD) was found either. Interestingly, 5 patients (4.95%) were infected with a CMV variant that lacked a signature RsaI digestion site determined by Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP), subsequent cloning and sequencing identified this CMV RsaI minus variant to be novel, herein designated as gB5. Overall, these findings suggest that CMV gB1 and gB3 are prevalent among HSCT recipients with CMV infections, and gB3 CMV infection is a risky indicator for CMV pneumonitis; furthermore, a novel CMV variant in a subset of Chinese HSCT recipients is identified and designated as gB5. PMID- 20025985 TI - Chemokine receptor CCR5 mediates alloimmune responses in graft-versus-host disease. AB - Allogeneic bone marrow transplantation (BMT) is an effective therapy for hematologic malignancies. However graft-versus-host disease (GVHD) is a major limiting factor for a successful patient outcome. GVHD is a result of alloimmune responses of donor T lymphocytes attacking the recipient's cells and tissues. Chemokine receptor CCR5 plays a role in solid organ allograft rejection and mediates murine GVHD pathogenesis. Herein, we report that infiltrating lymphocytes in the skin of human acute GVHD (aGVHD) samples are predominantly CCR5(+) T cells. In addition, we characterized the features of the CCR5 expression on alloreactive T lymphocytes. We found that the CCR5(+) population exhibits the characteristics of the activated effector T cell phenotype. CCR5 expression is upregulated upon allogenic stimulation, and CCR5(+) cells are proliferating with coexpression of T cell activation markers. Furthermore, the activated T cells producing inflammatory cytokine tumor necrosis factor (TNF)alpha, interleukin (IL)-2, or interferon (IFN)-gamma, are positive for CCR5. Thus, CCR5 is a marker for GVHD effector cells and CCR5(+) T cells are active participants in the pathogenesis of human aGVHD. PMID- 20025987 TI - Energetic requirements and bioenergetic modulation of mitochondrial morphology and dynamics. AB - Mitochondria are the site where oxidative phosphorylations (OXPHOSs) take place. Fusion and fission reactions allow them to change their overall morphology, which ranges from networks of elongated and branched filaments to collections of small individual organelles. It is assumed that mitochondrial bioenergetics and dynamics are linked and that mitochondrial morphology reflects their functional status. This review shows that the links between mitochondrial dynamics and bioenergetics are complex and that mitochondrial deficiencies are not systematically associated to fragmentation. In mammals, mitochondrial fragmentation is observed upon inhibition of OXPHOS with drugs, but not in most cellular models with OXPHOS deficits of genetic origin. In yeast, mitochondrial biogenesis and filament interconnectivity augment with increasing respiratory capacity, but mutation or inhibition of the respiratory chain does not provoke major morphological changes. Significant structural and morphological alterations appear restricted to mutation of genes involved in assembly or function of the F(1)F(0)-ATP-synthase. Finally, ex vivo studies (in mammals) and in vitro studies (in yeast) confirm the essential role of the inner membrane potential for mitochondrial fusion. PMID- 20025988 TI - The filamentous fungal gene expression database (FFGED). AB - Filamentous fungal gene expression assays provide essential information for understanding systemic cellular regulation. To aid research on fungal gene expression, we constructed a novel, comprehensive, free database, the filamentous fungal gene expression database (FFGED), available at http://bioinfo.townsend.yale.edu. FFGED features user-friendly management of gene expression data, which are assorted into experimental metadata, experimental design, raw data, normalized details, and analysis results. Data may be submitted in the process of an experiment, and any user can submit multiple experiments, thus classifying the FFGED as an "active experiment" database. Most importantly, FFGED functions as a collective and collaborative platform, by connecting each experiment with similar related experiments made public by other users, maximizing data sharing among different users, and correlating diverse gene expression levels under multiple experimental designs within different experiments. A clear and efficient web interface is provided with enhancement by AJAX (Asynchronous JavaScript and XML) and through a collection of tools to effectively facilitate data submission, sharing, retrieval and visualization. PMID- 20025989 TI - Long-term azithromycin use in patients with chronic obstructive pulmonary disease and tracheostomy. AB - Patients with Chronic Obstructive Pulmonary Disease (COPD) and tracheostomy are at high risk for exacerbations and hospitalizations. Macrolide treatment has shown to reduce exacerbations in moderate-to-severe COPD. To evaluate the safety and the efficacy of long-term azithromycin use in outpatients with severe COPD and tracheostomy. A multicenter, randomized, uncontrolled, pilot trial evaluating the safety and the efficacy of azithromycin 500 mg three day-a-week for 6 months (AZI) vs. standard of care (SC) in severe COPD outpatients with tracheostomy. Patients were monitored for six months of treatment plus six months of follow up. The primary outcome was the reduction in the number of exacerbations and hospitalizations. A total of 22 patients was randomized (11 to SC and 11 to AZI). Patients in AZI had a significant lower cumulative number of exacerbations after the first 3 months of treatment when compared to patients in SC (p = 0.001), as well as hospitalizations (p = 0.02). Kaplan-Meier survival curves for time to first exacerbation showed a significant reduction in AZI of the rates of first exacerbation when compared to SC (log rank test = 12.14, p < 0.001), as well as to first hospitalization (log-rank = 4.09, p = 0.04). Azithromycin significantly improved the quality of life in comparison to SC. No serious adverse events in the AZI group were reported. Long-term azithromycin treatment seems to be safe and effective in severe COPD outpatients with tracheostomy in reducing exacerbations, hospitalizations, as well as in improving quality of life. PMID- 20025990 TI - Acute cold-induced thermogenesis in neonatal chicks (Gallus gallus). AB - Mechanisms of thermogenic responses that play a role to maintain homeothermy during an early stage of neonatal chicks on acute cold exposure are scarcely studied as hatchlings are believed to be poikilothermic. However, chicks can attain the homeothermy during their subsequent growth after hatching. To identify thermogenic responses during neonatal stages of chicks (Gallus gallus) on acute cold exposure (12 degrees C, 3h), changes in plasma corticosterone levels, thermogenesis, gene transcripts (avUCP and avPGC1alpha) in skeletal muscles (pectoralis superficialis and gastrocnemius) and mitochondrial substrate oxidation enzyme activities in dissected tissues of 1- and 4-d-old chicks were studied. Results revealed that 1-day-old neonatal chicks were particularly vulnerable to cold exposure and were hypothermic. However, 4-d-old cold-exposed chicks maintained thermostability with significantly higher plasma corticosterone levels, oxygen consumption, heat production, and increased mitochondrial substrate oxidation enzyme activities (CS and 3HADH) in different dissected tissues. Analysis of gene transcripts for avian peroxisome-proliferator-activated receptor-gamma co-activator-1alpha (avPGC1alpha) and avian uncoupling protein (avUCP) in skeletal muscles revealed no significant change between cold-sensitive (1-d-old) and cold-tolerant (4-d-old) neonatal chicks, and failed to explain the enhanced thermogenesis and tolerance to acute cold. Additionally, avPGC1alpha gene transcripts were not correlated with the increased enzyme activities of CS or 3HADH in skeletal muscle. In conclusion, these data suggest the possible role of HPA-axis in cold-tolerant neonatal chicks to modulate substrate mobilization and oxidation. The thermogenic mechanisms based in part upon the increased capacity for mitochondrial substrate oxidation in different tissues are associated with enhanced heat production to attain homeothermy and acquire tolerance to acute cold exposure during an early stage of neonatal chicks. PMID- 20025991 TI - Scavenger receptors are associated with cellular interactions of S100A12 in vitro and in vivo. AB - Increased plasma levels of S100 proteins and interaction of S100 proteins with receptor for advanced glycation end products (RAGE) have been associated with a number of disease states, including chronic inflammatory processes and atherosclerosis. However, data concerning the role of circulating S100 proteins in these pathologies in vivo are scarce and, furthermore, it is currently not known whether RAGE is the sole receptor for extracellular S100 proteins in vivo. We report a novel methodology using recombinant human S100 proteins radiolabelled with fluorine-18, particularly, (18)F-S100A12, in receptor binding studies and cellular association studies in vitro, and in dynamic small animal positron emission tomography (PET) studies in rats in vivo. Association to both human aortic endothelial cells and macrophages revealed specific binding of (18)F S100A12 to RAGE, but, furthermore, provides evidence for interaction of (18)F S100A12 to various scavenger receptors (SR). PET data showed temporary association of (18)F-S100A12 with tissues overexpressing RAGE (e.g., lung), and, moreover, accumulation of (18)F-S100A12 in tissues enriched in cells overexpressing SR (e.g., liver and spleen). Blockade of overall SR interaction by maleylated BSA (malBSA) clearly shows diminished in vivo association of (18)F S100A12 to these tissues as well as a significant increment of the mean plasma residence time of (18)F-S100A12 (4.8+/-0.4 h vs. 2.3+/-0.3 h). The present approach first demonstrates that besides RAGE also scavenger receptors contribute to distribution, tissue association and elimination of circulating proinflammatory S100A12. PMID- 20025992 TI - Lipolysis in adipocytes. AB - Lipolysis in adipocytes, the hydrolysis of triacylglycerol (TAG) to release fatty acids (FAs) and glycerol for use by other organs, is a unique function of white adipose tissue. Lipolysis in adipocytes occurs at the surface of cytosolic lipid droplets, which have recently gained much attention as dynamic organelles integral to lipid metabolism. Desnutrin/ATGL is now established as a bona fide TAG hydrolase and mutations in human desnutrin/ATGL/PNPLA2, as well as in its activator, comparative gene identification 58, are associated with Neutral Lipid Storage Disease. Furthermore, recent identification of AdPLA as the major adipose phospholipase A(2), has led to the discovery of a dominant autocrine/paracrine regulation of lipolysis through PGE(2). Here, we review emerging concepts in the key players in lipolysis and the regulation of this process. We also examine recent findings in mouse models and humans with alterations/mutations in genes involved in lipolysis and discuss activation of lipolysis in adipocytes as a potential therapeutic target. PMID- 20025993 TI - The dietary flavonoids myricetin and fisetin act as dual inhibitors of DNA topoisomerases I and II in cells. AB - DNA topoisomerases (topos) are the target of several drugs commonly used in cancer chemotherapy; these drugs induce topo-DNA complexes with either topo I or topo II that eventually trigger cell death. The inhibition of these enzymes induces DNA alterations that may also lead to carcinogenic effects; indeed, an increased risk for developing leukemia has been observed in patients treated with some topo II inhibitors. Several flavonoids have been shown to interact with purified topo I and topo II, therefore suggesting that these compounds may possess both anticancer and carcinogenic activity. Because the activity of a drug on purified topoisomerases does not always represent the activity in the cell, the aim of this work is to evaluate the effects of several common dietary flavonoids on these enzymes in cells. Using the cell-based TARDIS assay, we have evaluated the effects of the flavonoids quercetin, apigenin, fisetin and myricetin on topo I and topo II in K562 human leukemia cells at several concentrations and exposure times. Quercetin and apigenin induced moderate levels of topo II-DNA complexes and did not induce topo I-DNA complexes in these cells. Fisetin induced neither topo I- nor topo II-DNA complexes, but behaved as a catalytic inhibitor of both enzymes. Myricetin induced high levels of topo-DNA complexes with both enzymes. In addition, murine embryo fibroblasts lacking topo IIbeta were resistant to myricetin-induced cell-growth inhibition, therefore suggesting that topo IIbeta is an important drug target for this flavonoid. These results support the idea that specific concentrations of some dietary flavonoids may produce topoisomerase-mediated carcinogenic and chemotherapeutic effects in vivo. The ability of myricetin to induce topo-DNA complexes with both topo I and topo II in leukemia cells may be therapeutically useful and deserves further study. PMID- 20025994 TI - A novel function of the human CLS1 in phosphatidylglycerol synthesis and remodeling. AB - Phosphatidylglycerol (PG) is a precursor for the biosynthesis of cardiolipin and a signaling molecule required for various cellular functions. PG is subjected to remodeling subsequent to its de novo biosynthesis in mitochondria to incorporate appropriate acyl content for its biological functions and to prevent the harmful effect of lysophosphatidylglycerol (LPG) accumulation. Yet, a gene encoding a mitochondrial LPG acyltransferase has not been identified. In this report, we identified a novel function of the human cardiolipin synthase (hCLS1) in regulating PG remodeling. In addition to the reported cardiolipin synthase activity, the recombinant hCLS1 protein expressed in COS-7 cells and Sf-9 insect cells exhibited a strong acyl-CoA-dependent LPG acyltransferase activity, which was further confirmed by purified hCLS1 protein overexpressed in Sf-9 cells. The recombinant hCLS1 displayed an acyl selectivity profile in the order of in the order of C18:1>C18:2>C18:0>C16:0, which is similar to that of hCLS1 toward PGs in cardiolipin synthesis, suggesting that the PG remodeling by hCLS1 is an intrinsic property of the enzyme. In contrast, no significant acyltransferase activity was detected from the recombinant hCLS1 enzyme toward lysocardiolipin which shares a similar structure with LPG. In support of a key function of hCLS1 in PG remodeling, overexpression of hCLS1 in COS-7 cells significantly increased PG biosynthesis concurrent with elevated levels of cardiolipin without any significant effects on the biosynthesis of other phospholipids. These results demonstrate for the first time that hCLS1 catalyzes two consecutive steps in cardiolipin biosynthesis by acylating LPG to PG and then converting PG to cardiolipin. PMID- 20025995 TI - Cloning and expression of ecdysone receptor (EcR) from the intertidal copepod, Tigriopus japonicus. AB - Ecdysteroids are steroid hormones that play an important role in development, growth, molting of larva, and reproduction in the Arthropoda. The effect of ecdysteroids is mediated by its binding to ecdysteroid receptor (EcR). To investigate the role of EcR during development and the effect to environmental stressors on EcR expression in a copepod, we isolated and characterized cDNA and 5'-promoter region of the Tigriopus japonicus EcR (TJ-EcR), and studied mRNA expression pattern. The full-length TJ-EcR cDNA sequence was 1962bp in length and the open reading frame encoded 546 amino acids. The deduced TJ-EcR protein contained well-conserved DNA-binding domain and ligand-binding domain. Phylogenetic analysis revealed that TJ-EcR was clustered with the EcR of other crustaceans. TJ-EcR mRNA was expressed in a developmental stage-specific manner: high in early developmental stages and low in the adult stage. Significantly elevated expression of the TJ-EcR gene in adults was detected at hypersalinity (42ppt) and high temperature (35 degrees C) condition. The 5'-flanking region of TJ-EcR gene contains heat shock protein 70 response elements, implying that the environmental stressors may affect its expression via the stress-sensor. In addition, bisphenol A (100microg/L) repressed TJ-EcR expression. Our results suggest that TJ-EcR could be a biomarker for the monitoring of the impact of environmental stressors in copepods. PMID- 20025997 TI - Hofmeister effects on activity and stability of alkaline phosphatase. AB - We have studied the effects on alkaline phosphatase of adding high concentrations (normally 1.0 M) of simple salts. It is necessary to allow for significant effects of salts on the extinction coefficient of the reaction product, and on the apparent pH of the buffer. Both activity and stability of the enzyme correlate well with the Hofmeister series in terms of the salt's kosmotropic/chaotropic properties, which are assessed by the Jones-Dole viscosity B coefficients (B(+) for cations and B(-) for anions). The catalytic activity or V(max)/K(m) of the enzyme showed a bell-shaped relationship with the (B(-)-B(+)) values of the salts present, being optimal with salts (such as NaCl, KCl, and KNO(3)) where the anion and cation have similar kosmotropic/chaotropic properties. This effect is believed to be enzyme-specific and relates to the impact of both cations and anions on the enzyme's surface pH, active site, and catalytic mechanism. Anions play a more predominant role than cations in affecting enzyme stability. The rate of irreversible thermal inactivation is strongly reduced by addition of kosmotropic anions like SO(4)(2-) (half-life increased from 8 to 580 min at 60 degrees C). This effect is general and the mechanism probably involves the ability of the ions to affect the water solvation layer around the enzyme molecule and to interact with both the surface and internal structure of the enzyme. PMID- 20025998 TI - Degradation of 316L stainless steel sternal wire by steam sterilization. AB - Sterilization is an important step prior to the implantation of medical devices inside the human body. In this work we studied the influence of steam sterilization cycles on the oxide film properties of stainless steel sternal wire. Characterization techniques such as open- circuit potential, potentiodynamic measurement, electrochemical impedance spectroscopy, cathodic stripping, transmission electron microscopy, atomic force microscopy and scanning electron microscopy were employed to investigate the cycles of steam sterilization on the corrosion behavior of sternal wire. The results showed that the oxide properties are a function of the number of steam sterilization cycles and deteriorate as the number of cycles increases. Steam sterilization might damage the implant integrity and heavy metals could be released to the surrounding tissues due to deterioration of the oxide film. PMID- 20025999 TI - Synthesis and characterization of hyaluronic acid-poly(ethylene glycol) hydrogels via Michael addition: An injectable biomaterial for cartilage repair. AB - Injectable hydrogels based on hyaluronic acid (HA) and poly(ethylene glycol) (PEG) were designed as biodegradable matrices for cartilage tissue engineering. Solutions of HA conjugates containing thiol functional groups (HA-SH) and PEG vinylsulfone (PEG-VS) macromers were cross-linked via Michael addition to form a three-dimensional network under physiological conditions. Gelation times varied from 14min to less than 1min, depending on the molecular weights of HA-SH and PEG VS, degree of substitution (DS) of HA-SH and total polymer concentration. When the polymer concentration was increased from 2% to 6% (w/v) in the presence of 100Uml(-1) hyaluronidase the degradation time increased from 3 to 15days. Hydrogels with a homogeneous distribution of cells were obtained when chondrocytes were mixed with the precursor solutions. Culturing cell-hydrogel constructs prepared from HA185k-SH with a DS of 28 and cross-linked with PEG5k 4VS for 3weeks in vitro revealed that the cells were viable and that cell division took place. Gel-cell matrices degraded in approximately 3weeks, as shown by a significant decrease in dry gel mass. At day 21 glycosaminoglycans and collagen type II were found to have accumulated in hydrogels. These results indicate that these injectable hydrogels have a high potential for cartilage tissue engineering. PMID- 20026000 TI - Screening of peptides with a high affinity for ZnO using spot-synthesized peptide arrays and computational analysis. AB - Metal-binding peptides have attracted attention for their usefulness in biomineralization processes. In this work screening of high affinity peptides against ZnO was performed using a combinatorial library approach with a peptide array and computational analysis. The computationally assisted peptide screening and design enabled identification of linear peptides with affinity for ZnO with limited experimentation. Starting with the screening data obtained from a random 6-mer library of 420 sequences, the characteristics of the peptides with high ZnO affinity were analyzed using a fuzzy neural network algorithm by comparison of high and low affinity peptides. Three physical properties of amino acids (hydrophobicity, isoelectric point and size) and positional information of each residue were analyzed and a peptide rule with restricted amino acids at certain positions was extracted. The average affinity for ZnO increased 2.0-fold when 300 sequences were synthesized according to the restricted random library, compared with the non-restricted library. In addition, for a peptide library with the amino acids at the restricted sites exchanged the average binding capacity decreased to 0.7-fold. Interestingly, the peptides with high ZnO affinity obtained exhibited binding specificity for ZnO. Computationally assisted screening is an invaluable means for finding peptides with limited experimentation. PMID- 20026002 TI - Bariatric surgery. AB - Obesity is fast becoming one of the world's leading health problems and together with its many associated medical sequelae significantly increases morbidity and mortality. In this review, we briefly explore the history of bariatric surgery, the benefits of surgery and the various procedures carried out. PMID- 20026001 TI - Peri-operative antibiotic prophylaxis: adherence to guidelines and effects of educational intervention. AB - BACKGROUND: This study was aimed to determine the propriety of perioperative antimicrobial prophylaxis, to evaluate the effects of a training intervention on correction. METHODS: This prospective educational intervention study was performed on clean, clean-contaminated, and contaminated wounds in elective procedures. The accuracy of the antimicrobial prophylaxis was analyzed according to international guidelines. The outcome measures were appropriateness of prophylactic antibiotic indication, choice, dose and duration; and cost of inappropriate administration. RESULTS: Before the intervention, 312 procedures were recorded compared with 322 after the intervention. Total compliance rate decreased from 34.3% to 28.5% after the intervention, though insignificantly (p=0.59). Educational training intervention did not change the rate of inappropriate antibiotic choice statistically, and prolonged antibiotic use was significantly higher after the intervention (p=0.01). The positive impact of the intervention was observed in decreased "not indicated but administered" rate (p=0.009) and absence of procedures with "indicated but not administered" and "inappropriate antibiotic dose" which also decreased significantly (both p<0.001). The total cost of irrational antibiotic use was US$26,230.20. CONCLUSIONS: Although this educational intervention study achieved improvements in indications, choice, and dosing of surgical antimicrobial prophylaxis, it failed to improve prolonged use and total compliance rate, and to lower the costs sufficiently. It will probably provide better results by means of compulsory measures for surgeons to comply with available protocols and guidelines, as well as education programs, in order to change the antibiotic utility habits on surgical prophylaxis and to achieve accurate prophylactic administrations. PMID- 20026003 TI - Proteome of Gluconacetobacter diazotrophicus co-cultivated with sugarcane plantlets. AB - Gluconacetobacter diazotrophicus is a micro-aerobic bacterium able to fix atmospheric nitrogen in endophytic mode. A proteomic approach was used to analyze proteins differentially expressed in the presence and absence of sugarcane plantlets. Two-dimensional gel electrophoresis (2-DE) showed 42 spots with altered levels of expression. Analysis of these spots by matrix-assisted laser desorption ionization time-of-flight in tandem (MALDI-TOF-TOF) identified 38 proteins. Differentially expressed proteins were associated with carbohydrate and energy metabolism, folding, sorting and degradation processes, and transcription and translation. Among proteins expressed in co-cultivated bacteria, four belong to membrane systems; others, like a transcription elongation factor (GreA), a 60 kDa chaperonin (GroEL), and an outer membrane lipoprotein (Omp16) have also been described in other plant-bacteria associations, indicating a common protein expression pattern as a result of symbiosis. A high protein content of 60kDa chaperonin isoforms was detected as non-differentially expressed proteins of the bacteria proteome. These results allow the assessment of the physiological significance of specific proteins to G. diazotrophicus metabolism and to the pathways involved in bacteria-host endophytic interaction. PMID- 20026004 TI - Protein profile in HBx transfected cells: a comparative iTRAQ-coupled 2D LC-MS/MS analysis. AB - The x protein of HBV (HBx) has been involved in the development of hepatocellular carcinoma (HCC), with a possible link to individual genotypes. Nevertheless, the underlying mechanism remains obscure. In this study, we aim to identify the HBx induced protein profile in HepG2 cells by LC-MS/MS proteomics analysis. Our results indicated that proteins were differentially expressed in HepG2 cells transfected by HBx of various genotypes. Proteins associated with cytoskeleton were found to be either up-regulated (MACF1, HMGB1, Annexin A2) or down-regulated (Lamin A/C). These may in turn result in the decrease of focal adhesion and increase of cell migration in response to HBx. Levels of other cellular proteins with reported impact on the function of extracellular matrix (ECM) proteins and cell migration, including Ca(2+)-binding proteins (S100A11, S100A6, and S100A4) and proteasome protein (PSMA3), were affected by HBx. The differential protein profile identified in this study was also supported by our functional assay which indicated that cell migration was enhanced by HBx. Our preliminary study provided a new platform to establish a comprehensive cellular protein profile by LC-MS/MS proteomics analysis. Further downstream functional assays, including our reported cell migration assay, should provide new insights in the association between HCC and HBx. PMID- 20026005 TI - A long-wavelength fluorescent substrate for continuous fluorometric determination of alpha-mannosidase activity: resorufin alpha-D-mannopyranoside. AB - A simple and reliable continuous assay for measurement of alpha-mannosidase activity is described and demonstrated for analysis with two recombinant human enzymes using the new substrate resorufin alpha-d-mannopyranoside (Res-Man). The product of enzyme reaction, resorufin, exhibits fluorescence emission at 585 nm with excitation at 571 nm and has a pK(a) of 5.8, allowing continuous measurement of fluorescence turnover at or near physiological pH values for human lysosomal and Drosophila Golgi alpha-mannosidases. The assay performed using recombinant Drosophila Golgi alpha-mannosidase (dGMII) has been shown to give the kinetic parameters K(m) of 200 microM and V(max) of 11 nmol/min per nmol dGMII. Methods for performing the assay using several concentrations of the known alpha mannosidase inhibitor swainsonine are also presented, demonstrating a potential for use of the assay as a simple method for high-throughput screening of inhibitors potentially useful in cancer treatment. PMID- 20026006 TI - Cyclophilin D in mitochondrial pathophysiology. AB - Cyclophilins are a family of peptidyl-prolyl cis-trans isomerases whose enzymatic activity can be inhibited by cyclosporin A. Sixteen cyclophilins have been identified in humans, and cyclophilin D is a unique isoform that is imported into the mitochondrial matrix. Here we shall (i) review the best characterized functions of cyclophilin D in mitochondria, i.e. regulation of the permeability transition pore, an inner membrane channel that plays an important role in the execution of cell death; (ii) highlight new regulatory interactions that are emerging in the literature, including the modulation of the mitochondrial F1FO ATP synthase through an interaction with the lateral stalk of the enzyme complex; and (iii) discuss diseases where cyclophilin D plays a pathogenetic role that makes it a suitable target for pharmacologic intervention. PMID- 20026007 TI - Knockdown of F1 epsilon subunit decreases mitochondrial content of ATP synthase and leads to accumulation of subunit c. AB - The subunit epsilon of mitochondrial ATP synthase is the only F1 subunit without a homolog in bacteria and chloroplasts and represents the least characterized F1 subunit of the mammalian enzyme. Silencing of the ATP5E gene in HEK293 cells resulted in downregulation of the activity and content of the mitochondrial ATP synthase complex and of ADP-stimulated respiration to approximately 40% of the control. The decreased content of the epsilon subunit was paralleled by a decrease in the F1 subunits alpha and beta and in the Fo subunits a and d while the content of the subunit c was not affected. The subunit c was present in the full-size ATP synthase complex and in subcomplexes of 200-400 kDa that neither contained the F1 subunits, nor the Fo subunits. The results indicate that the epsilon subunit is essential for the assembly of F1 and plays an important role in the incorporation of the hydrophobic subunit c into the F1-c oligomer rotor of the mitochondrial ATP synthase complex. PMID- 20026008 TI - Chloroplastic oxidative burst induced by tenuazonic acid, a natural photosynthesis inhibitor, triggers cell necrosis in Eupatorium adenophorum Spreng. AB - Tenuazonic acid (TeA), a nonhost-specific phytotoxin produced by Alternaria alternata, was determined to be a novel natural photosynthesis inhibitor owning several action sites in chloroplasts. To further elucidate the mode of its action, studies were conducted to assess the production and involvement of reactive oxygen species (ROS) in the toxic activity of TeA. A series of experiments indicated that TeA treatment can induce chloroplast-derived ROS generation including not only (1)O(2) but also superoxide radical, H(2)O(2) and hydroxyl radicals in Eupatorium adenophorum mesophyll cells, resulting from electron leakage and charge recombination in PSII as well as thylakoid overenergization due to inhibition of the PSII electron transport beyond Q(A) and the reduction of end acceptors on the PSI acceptor side and chloroplast ATPase activity. The initial production of TeA-induced ROS was restricted to chloroplasts and accompanied with a certain degree of chloroplast damage. Subsequently, abundant ROS were quickly dispersed throughout whole cell and cellular compartments, causing a series of irreversible cellular harm such as chlorophyll breakdown, lipid peroxidation, plasma membrane rupture, chromatin condensation, DNA cleavage, and organelle disintegration, and finally resulting in rapid cell destruction and leaf necrosis. These results show that TeA causing cell necrosis of host-plants is a result of direct oxidative damage from chloroplast-mediated ROS eruption. PMID- 20026009 TI - Redox potential of the Rieske iron-sulfur protein quantum-chemical and electrostatic study. AB - Quantum-chemical study of structures, energies, and effective partial charge distribution for several models of the Rieske protein redox center is performed in terms of the B3LYP density functional method in combination with the broken symmetry approach using three different atomic basis sets. The structure of the redox complex optimized in vacuum differs markedly from that inside the protein. This means that the protein matrix imposes some stress on the active site resulting in distortion of its structure. The redox potentials calculated for the real active site structure are in a substantially better agreement with the experiment than those calculated for the idealized structure. This shows an important role of the active site distortion in tuning its redox potential. The reference absolute electrode potential of the standard hydrogen electrode is used that accounts for the correction caused by the water surface potential. Electrostatic calculations are performed in the framework of the polarizable solute model. Two dielectric permittivities of the protein are employed: the optical permittivity for calculation of the intraprotein electric field, and the static permittivity for calculation of the dielectric response energy. Only this approach results in a reasonable agreement of the calculated and experimental redox potentials. PMID- 20026010 TI - Mitochondrial uncoupling proteins in unicellular eukaryotes. AB - Uncoupling proteins (UCPs) are members of the mitochondrial anion carrier protein family that are present in the mitochondrial inner membrane and mediate free fatty acid (FFA)-activated, purine nucleotide (PN)-inhibited proton conductance. Since 1999, the presence of UCPs has been demonstrated in some non photosynthesising unicellular eukaryotes, including amoeboid and parasite protists, as well as in non-fermentative yeast and filamentous fungi. In the mitochondria of these organisms, UCP activity is revealed upon FFA-induced, PN inhibited stimulation of resting respiration and a decrease in membrane potential, which are accompanied by a decrease in membranous ubiquinone (Q) reduction level. UCPs in unicellular eukaryotes are able to divert energy from oxidative phosphorylation and thus compete for a proton electrochemical gradient with ATP synthase. Our recent work indicates that membranous Q is a metabolic sensor that might utilise its redox state to release the PN inhibition of UCP mediated mitochondrial uncoupling under conditions of phosphorylation and resting respiration. The action of reduced Q (QH2) could allow higher or complete activation of UCP. As this regulatory feature was demonstrated for microorganism UCPs (A. castellanii UCP), plant and mammalian UCP1 analogues, and UCP1 in brown adipose tissue, the process could involve all UCPs. Here, we discuss the functional connection and physiological role of UCP and alternative oxidase, two main energy-dissipating systems in the plant-type mitochondrial respiratory chain of unicellular eukaryotes, including the control of cellular energy balance as well as preventive action against the production of reactive oxygen species. PMID- 20026011 TI - The role of beta(1)Pix/caveolin-1 interaction in endothelin signaling through Galpha subunits. AB - Endothelin-1 (ET-1) is a potent mitogen that transmits signals through its cognate G protein-coupled receptors to stimulate extracellular signal-regulated kinase Erk1/2. Endothelin-1 receptors (ET-Rs) are known to interact with caveolin 1 and co-localize in caveolae which integrate different receptor and signaling proteins. We have recently shown that beta(1)Pix binds specifically to ET-Rs. Here, we show that beta(1)Pix binding to caveolin-1 is dependent on heterotrimeric G proteins activation state. beta(1)Pix interaction with different G proteins is increased in the presence of the G protein activator AMF. Moreover, extraction of cholesterol with methyl-beta-cyclodextrin disrupts the binding of beta(1)Pix to Galpha(q), Galpha(12) and phospho-Erk1/2 but not the binding of beta(1)Pix to G(beta1). The disruption of beta(1)Pix dimerization strongly reduced the binding of caveolin-1, Galpha(q) and Galpha(12). Constitutively active mutants of Galpha(q) and Galpha(12) increased Cdc42 activation when co expressed with beta(1)Pix but not in the presence of beta(1)Pix dimerization deficient mutant beta(1)PixDelta (602-611). ET-1 stimulation increased the binding of phosphorylated Erk1/2 to beta(1)Pix but not to beta(1)PixDelta (602 611). RGS3 decreased ET-1-induced Cdc42 activation. These results strongly suggest that the activation of ET-Rs leads to the compartmentalization and the binding of Galpha(q) to beta(1)Pix in caveolae, where dimeric beta(1)Pix acts as platform to facilitate the binding and the activation of Erk1/2. PMID- 20026012 TI - Downregulation of Gnas, Got2 and Snord32a following tenofovir exposure of primary osteoclasts. AB - Clinical observations have implicated the antiretroviral drug tenofovir with bone density loss during the management of HIV infection. The goal of this study was to investigate the in vitro effects of tenofovir exposure of primary osteoclasts in order to gain insights into the potential mechanisms for the drug-induced bone density loss. We hypothesized that tenofovir may alter the expression of key genes involved in osteoclast function. To test this, primary osteoclasts were exposed to physiologically relevant concentrations of the prodrug tenofovir disoproxil fumarate (TDF), then intensive microarray analysis was done to compare tenofovir-treated versus untreated cells. Specific downregulation of Gnas, Got2 and Snord32a were observed in the TDF-treated cells. The functions of these genes help to explain the basis for tenofovir-associated bone density loss. Our studies represent the first analysis of the effects of tenofovir on osteoclast gene expression and help to explain the basis of tenofovir-associated bone density loss in HIV-infected individuals. PMID- 20026013 TI - Identification of candidate genes involved in endogenous protection mechanisms against acute pancreatitis in mice. AB - We surveyed changes of the gene expression profile in caerulein-exposed pancreas using Affymetrix GeneChip system (39,000 genes). Up-regulation of genes coding for claudin 4, claudin 7, F11 receptor, cadherin 1, integrin beta 4, syndecan 1, heat shock proteins b1/90aa1, Serpinb6a, Serpinb6b, Serpinb9, Bax, Bak1, calpain 2, calpain 5, microtubule-associated protein 1 light chain 3 alpha, S100 calcium binding proteins A4/A10 were found in mouse pancreas exposed to caerulein for 12h. In contrast, the anti-apoptotic gene Bcl2 was down-regulated. The functions of these genes concern tight junction formation, cell-cell/cell-matrix adhesions, stress response, protease inhibition, apoptosis, autophagy, and regulation of cytoskeletal dynamics. Caerulein-exposed pancreatic acinar cells were immunohistochemically stained for claudin 4, cadherin 1, integrin beta 4, heat shock protein b1, and Serpinb6a. In conclusion, we have newly identified a set of genes that are likely to be involved in endogenous self-protection mechanisms against acute pancreatitis. PMID- 20026014 TI - Corticosterone basal levels and vulnerability to LPS-induced neuroinflammation in the rat brain. AB - To assess whether the individual differences on the brain response to lipopolysaccharide (LPS) are correlated with the individual differences in the hypothalamus-pituitary-adrenal axis basal activity, adult male outbred rats were injected i.p. with 1 mg/kg LPS and evaluated after 4 h. Basal (1 week before LPS) and post-LPS plasma corticosterone (CC) were measured (mean basal: 225+/-22 ng/mL at 15:00 h). Group H was assigned to animals with 33% higher levels of CC (>234 ng/mL) and group L to animals with 33% lower levels of CC (<167 ng/mL). The H group showed an 8.8 times less relative increase of CC after LPS than the L group as well as a reduced glucocorticoid receptor upregulation after LPS. In addition, H individuals present higher plasma levels of TNF-alpha and IL-1beta after LPS. Interestingly, these animals are more vulnerable to the accumulation of oxidative/nitrosative mediators in the brain (NF-kappaB, NOS-2 and COX-2). Concomitantly, H animals are less protected against LPS-induced neuroinflammation, since anti-inflammatory mediators, lipocalin-prostaglandinD2 synthase and peroxisome proliferator-activated gamma, are downregulated after LPS. These data demonstrate that CC plasma basal levels might be a relevant parameter for predicting the individual response to LPS. PMID- 20026015 TI - Effects of triterpene derivatives from Maytenus rigida on VEGF-induced Kaposi's sarcoma cell proliferation. AB - Betulinic acid (BA) is a naturally occurring lupane-type triterpene which exhibits a variety of biological activities including potent cytotoxic properties. On the basis of the structural similarity to BA, two lupane derivatives namely lup-20(29)-ene-3beta,30-diol (1) and lup-20(29)-ene-3beta,28 diol (2), along with two friedelane derivatives, namely friedelan-3-one (3) and friedelan-3beta-ol (4), isolated from the Brazilian plant Maytenus rigida, have been evaluated for their anti-proliferative effect. Similarly to BA, compounds 1 and 3 at 1 microM concentration significantly inhibited the VEGF-induced Kaposi's sarcoma (KS) cell proliferation by 50%. In contrast, this effect was not found in control endothelial cells (EC). Moreover, compounds 1 and 3 showed a dose dependent effect on the apoptotic cell death, as detected by FACS analysis and caspase-3 assay. Specifically, at 10 microM concentration, apoptosis was significantly induced (from 45% to 55% of hypodiploid cells vs control cells) and showed the same potency order observed for the anti-proliferative effect at 1 microM, i.e., compound 3>BA>compound 1. Taking into account the interest given rise by BA as anticancer agent, the comparable anti-proliferative activity shown by compounds 1 and 3 and BA, can give an impulse to further investigate lupane and friedelane derivatives as cytotoxic agents. PMID- 20026016 TI - Implications of latency period between benzene exposure and development of leukemia--a synopsis of literature. AB - From numerous epidemiological studies it is evidenced that risk after exposure to a carcinogen varies with time and there has been an increasing discussion about the temporal variation in case of smoking, ionizing radiation and various chemical carcinogens. The results of several independent epidemiologic studies of occupational cohorts with benzene exposure and the development of leukemia will be presented in order to find common aspects. In this context the data of 537 confirmed cases of leukemia as an occupational disease in Germany during the time period 1978-2007 will be presented. It is concluded that the epidemiologic findings are consistent and demonstrate a smaller or even absent risk of leukemia 10-15 years after exposure to benzene has been stopped. Temporal changes in relative risk highlights the importance of examining the relationship between follow-up time and risk estimates as part of the risk assessment process. PMID- 20026017 TI - Incidence and susceptibility to therapy-related myeloid neoplasms. AB - Therapy-related myeloid neoplasms (t-MN) include acute myeloid leukemias and myelodysplastic syndromes arising in patients who have been treated with chemotherapy, radiation therapy, immunosuppressive agents or after documented exposure to environmental carcinogen. t-MN are defined according to the primary treatment and the corresponding genetic and molecular lesions. Chromosome(s) 7 and/or 5 monosomies or deletions are typical of alkylating agent-induced AML, while balanced translocations involving chromosome bands 11q23 and 21q22 are associated to preceeding therapy with DNA-topoisomerase II inhibitors. Antimetabolites, and in particular the immunosuppressive agents azathioprine and fludarabine, have also been recently associated to t-MN. Leukemias developing after benzene exposure are similar to t-MN and are characterized by chromosomal aberrations, which have been also observed among otherwise healthy benzene exposed workers. Individual predisposing factors, including polymorphisms of detoxification and DNA-repair enzymes have been identified. Two genetic variants in key metabolizing enzymes, myeloperoxidase and NAD(P)H:quinone oxidoreductase, have been shown to influence susceptibility to benzene hematotoxicity. Combination of polymorphisms impairing detoxification and DNA repair may significantly increase therapy-related myeloid neoplasm risk. Among hematological malignancies, long-term survivors of Hodgkin's lymphoma are exposed to an increased t-MN risk, particularly when receiving MOPP-based and escalated-BEACOPP regimens, and when alkylators are combined to radiotherapy. Patients with lymphoma are at highest risk if total body irradiation followed by autologous stem cell transplantation is used as rescue or consolidation. The addition of granulocyte-colony stimulating factor (G-CSF) and radiotherapy plays a significant role in t-MN following treatment of childhood acute lymphoblastic leukemia. In solid tumors, treatment for breast cancer and germ-cell tumors has been associated with a 1-5% lifetime risk of t-MN. PMID- 20026018 TI - Detection of serum beta(2)-GPI-Lp(a) complexes in patients with systemic lupus erythematosus. AB - BACKGROUND: Circulating beta(2)-glycoprotein-I-oxidized low-density lipoprotein (beta(2)-GPI-ox-LDL) complexes have been found in patients with systemic lupus erythematosus (SLE) and other autoimmune diseases as a contributor to the development of autoimmune-mediated atherosclerosis. In vitro study showed that beta(2)-GPI also bound with high affinity to atherogenic lipoprotein (a) [Lp(a)] which shares structural similarity to LDL. We examined the existence and clinical significance of serum complexes of beta(2)-GPI with Lp(a) in SLE patients. METHODS: A "sandwich" ELISA was developed for measuring serum concentrations of beta(2)-GPI-Lp(a) complexes, using rabbit anti-human beta(2)-GPI antibody as capturing antibody, and quantitating with antibody against apo(a). Forty-seven SLE patients and 42 healthy controls were studied. RESULTS: Both Lp(a) (400+/-213 mg/l vs. 181+/-70 mg/l) and ox-Lp(a) (27.07+/-22.30 mg/l vs. 8.20+/-4.55 mg/l) concentrations were higher in SLE patients than in controls (P<0.0001). beta(2) GPI-Lp(a) complexes were detectable in both controls and SLE. The complexes levels in SLE were higher than in controls (0.96+/-0.41 U/ml vs. 0.59+/-0.20 U/ml, P<0.0001) and was positively correlated with ox-Lp(a) (P<0.001). CONCLUSIONS: We report the existence of beta(2)-GPI-Lp(a) complexes in both controls and SLE patients. The complexes levels increase in SLE. PMID- 20026019 TI - Prevalence of gastric parietal cell antibodies and intrinsic factor antibodies in primary biliary cirrhosis. AB - BACKGROUND: We investigated the prevalence of antibodies against gastric parietal cells (GPA), intrinsic factor antibodies (IFA) and the presence of pernicious anemia in a large cohort of primary biliary cirrhosis (PBC) patients as similar data is missing. METHODS: 157 PBC patients and 357 controls (73 with autoimmune hepatitis (AIH), 35 primary sclerosing cholangitis (PSC), 45 HBV, 37 HCV, 36 alcoholic liver disease (ALD), 35 non-alcoholic fatty liver disease (NAFLD) and 96 healthy) were investigated for IgG-isotype-specific GPA and IFA by ELISAs and vitamin-B(12) levels by a microparticle enzyme immunoassay. RESULTS: The detection of IgG-GPA was significantly higher in PBC (31.8%) compared to AIH (10.9%; p=0.001), PSC (0%; p=0.000), HCV (13.5%; p=0.01), HBV (13.3%; p=0.006), ALD (8.3%; p=0.004), NAFLD (11.4%; p=0.003) and healthy (10.4%; p=0.001). IgG-IFA were detected in 12% of GPA-positive PBC patients and in none of the other liver diseases or in healthy (p=0.001). This reactivity was significantly associated with lower vitamin-B(12) levels compared to those with an IFA-negative test (p=0.025). CONCLUSIONS: A significant proportion of PBC patients had IgG-GPA and IFA compared to controls. IgG-IFA were detected only in GPA-positive PBC patients and associated with lower vitamin-B(12) levels compared to those with an IFA negative test. PMID- 20026020 TI - Evaluation of the ARCHITECT urine NGAL assay: assay performance, specimen handling requirements and biological variability. AB - OBJECTIVES: NGAL (Neutrophil Gelatinase-Associated Lipocalin) has emerged as a new biomarker for the identification of acute kidney injury. Reliable clinical evaluations require a simple, robust test method for NGAL, and knowledge of specimen handling and specimen stability characteristics. We evaluated the performance of a new urine NGAL assay on the ARCHITECT analyzer. METHODS: Assay performance characteristics were evaluated using standard protocols. Urine specimen storage requirements were determined and biological variability was assessed in a self-declared apparently healthy population. RESULTS: Assay performance data showed good precision, sensitivity and lot-to-lot reproducibility. There was good short term 2-8 degrees C sample stability, however, long term storage samples must be kept at -70 degrees C or colder. The largest variance component in a biological variance study was within-day. CONCLUSIONS: The ARCHITECT NGAL assay proved to be a precise and reproducible assay for the determination of urine NGAL. PMID- 20026021 TI - Screening for inborn errors of metabolism using automated electrospray tandem mass spectrometry: study in high-risk Indian population. AB - OBJECTIVES: Tandem mass spectrometry is a major technological advance in the screening for inborn errors of metabolism. It has the advantage of sensitive and simultaneous multiple disease screening with minimal sample requirement. The diseases detected include aminoacidemias, fatty acid oxidation disorders, and organic acidemias. DESIGN AND METHODS: Using automated electrospray tandem mass spectrometry we screened 3550, clinically selected, symptomatic children for inborn errors of metabolism by analyzing amino acids and acylcarnitines in dried blood filter-paper samples. RESULTS: Among these, 113 (3.2%) children were identified with a metabolic disorder: 61 (54%) patients had amino acid disorders, 47 (41.6%) had organic acidemias, and 5 (4.4%) children had disorders of fatty acid oxidation. The diagnoses were further confirmed through clinical symptoms, and other biochemical studies. CONCLUSIONS: These results show that inherited metabolic disorders are not rare in India, a rapidly developing country with a high birth rate and relatively frequent occurrence of consanguineous marriages. PMID- 20026022 TI - Bisalbuminuria detected by agarose gel electrophoresis. PMID- 20026023 TI - Direct measurement of serum free testosterone by ultrafiltration followed by liquid chromatography tandem mass spectrometry. AB - BACKGROUND: Currently there is no reliable method suitable for routine measurement of serum free testosterone (FT). AIM: To develop such a method involving liquid chromatography tandem mass spectrometry (LC-IDMS/MS) that directly detects and quantifies the FT present in serum. METHODS: Ultrafiltrate testosterone obtained from 0.5 mL of serum was partially purified by liquid/liquid extraction and quantified using an Agilent 1200 Series HPLC system coupled to an API 5000 mass spectrometer equipped with an atmospheric pressure chemical ionization ion source. Using split samples serum free testosterone was compared between direct ultrafiltration (UF) coupled LC-MS/MS, analogue FT immunoassay, free testosterone calculated from mass action equations (cFT) and with equilibrium dialysis (ED) coupled LC-MS/MS. RESULTS: Total imprecision determined over twenty runs was <6% at 67 pmol/L and 158 pmol/L FT. The dynamic response was linear up to at least 2500 pmol/L while physical LLOQ (18 % CV) equaled 16 pmol/L. The UF method agreed poorly with analogue immunoassay (correlation coefficient 0.667; bias -81%), somewhat better against cFT when total testosterone was determined by immunoassay (correlation coefficient 0.816, bias 21% ) and still better yet against cFT when total testosterone was determined by LC-MS/MS (correlation coefficient 0.8996, bias 10%). Agreement was closest with ED method (correlation coefficient 0.9779, bias 2.4%). CONCLUSION: We present a relatively simple UF coupled LC-MS/MS definitive method that measures serum free testosterone. The method is relatively fast, reliable and is suitable for the routine clinical laboratory practice. PMID- 20026024 TI - hunchback and Ikaros-like zinc finger genes control reproductive system development in Caenorhabditis elegans. AB - Here we provide evidence for a C2H2 zinc finger gene family with similarity to Ikaros and hunchback. The founding member of this family is Caenorhabditis elegans ehn-3, which has important and poorly understood functions in somatic gonad development. We examined the expression and function of four additional hunchback/Ikaros-like (HIL) genes in C. elegans reproductive system development. Two genes, ehn-3 and R08E3.4, are expressed in somatic gonadal precursors (SGPs) and have overlapping functions in their development. In ehn-3; R08E3.4 double mutants, we find defects in the generation of distal tip cells, anchor cells, and spermatheca; three of the five tissues derived from the SGPs. We provide in vivo evidence that C. elegans HIL proteins have functionally distinct zinc finger domains, with specificity residing in the N-terminal set of four zinc fingers and a likely protein-protein interaction domain provided by the C-terminal pair of zinc fingers. In addition, we find that a chimeric human Ikaros protein containing the N-terminal zinc fingers of EHN-3 functions in C. elegans. Together, these results lend support to the idea that the C. elegans HIL genes and Ikaros have similar functional domains. We propose that hunchback, Ikaros, and the HIL genes arose from a common ancestor that was present prior to the divergence of protostomes and deuterostomes. PMID- 20026025 TI - Giant, Kruppel, and caudal act as gap genes with extensive roles in patterning the honeybee embryo. AB - In Drosophila, gap genes translate positional information from gradients of maternal coordinate activity and act to position the periodic patterns of pair rule gene stripes across broad domains of the embryo. In holometabolous insects, maternal coordinate genes are fast-evolving, the domains that gap genes specify often differ from their orthologues in Drosophila while the expression of pair rule genes is more conserved. This implies that gap genes may buffer the fast evolving maternal coordinate genes to give a more conserved pair-rule output. To test this idea, we have examined the function and expression of three honeybee orthologues of gap genes, Kruppel, caudal, and giant. In honeybees, where many Drosophila maternal coordinate genes are missing, these three gap genes have more extensive domains of expression and activity than in other insects. Unusually, honeybee caudal mRNA is initially localized to the anterior of the oocyte and embryo, yet it has no discernible function in that domain. We have also examined the influence of these three genes on the expression of honeybee even-skipped and a honeybee orthologue of engrailed and show that the way that these genes influence segmental patterning differs from Drosophila. We conclude that while the fundamental function of these gap genes is conserved in the honeybee, shifts in their expression and function have occurred, perhaps due to the apparently different maternal patterning systems in this insect. PMID- 20026027 TI - Electrophysiological characterization of harmane-induced activation of mesolimbic dopamine neurons. AB - It has been suggested that the beta-carbolines harmane and norharmane may be involved in the pathophysiology of Parkinson's disease, psychosis and addiction, but the mechanisms of these possible effects remain to be elucidated. In the present study, the effects of the two compounds were examined by using in vivo extracellular recordings of ventral tegmental dopamine neurons. The effects of harmane (2mg/kg) and norharmane (2mg/kg), were compared to those of nicotine (11microg/kg), of cotinine (0.5mg/kg), of the monoamine-oxidase-A inhibitor befloxatone (0.12mg/kg), and of the monoamine-oxidase-B inhibitor selegiline (0.5mg/kg). The effects of harmane were also tested after pre-treatment with the nicotine receptor antagonist mecamylamine. The results show that all substances, except befloxatone, activate the firing and/or burst activity of dopamine neurons. The increase in firing rate produced by harmane was approximately 18 times greater than that produced by nicotine. Such powerful excitation of dopamine neurons by harmane may in part explain its involvement in neurotoxicity, psychosis and addiction. The absence of effect of befloxatone supports the hypothesis that the effect of harmane is not related to its monoamine-oxidase-A inhibitory properties. Mecamylamine inhibited by approximately 80% the activity of harmane, indicating that the activating effect of harmane on dopamine neurons involves several mechanisms, among which activation of nicotinic receptors likely has a prominent importance. The results of the present study support the hypothesis that harmane could be a tobacco (or smoke) component other than nicotine involved in tobacco dependence. PMID- 20026026 TI - Long-range neural and gap junction protein-mediated cues control polarity during planarian regeneration. AB - Having the ability to coordinate the behavior of stem cells to induce regeneration of specific large-scale structures would have far-reaching consequences in the treatment of degenerative diseases, acute injury, and aging. Thus, identifying and learning to manipulate the sequential steps that determine the fate of new tissue within the overall morphogenetic program of the organism is fundamental. We identified novel early signals, mediated by the central nervous system and 3 innexin proteins, which determine the fate and axial polarity of regenerated tissue in planarians. Modulation of gap junction dependent and neural signals specifically induces ectopic anterior regeneration blastemas in posterior and lateral wounds. These ectopic anterior blastemas differentiate new brains that establish permanent primary axes re-established during subsequent rounds of unperturbed regeneration. These data reveal powerful novel controls of pattern formation and suggest a constructive model linking nervous inputs and polarity determination in early stages of regeneration. PMID- 20026028 TI - Role of the 207-218 peptide region of Moloney murine leukemia virus integrase in enzyme catalysis. AB - X-ray diffraction data on a few retroviral integrases show a flexible loop near the active site. By sequence alignment, the peptide region 207-218 of Mo-MLV IN appears to correspond to this flexible loop. In this study, residues H208, Y211, R212, Q214, S215 and S216 of Mo-MLV IN were mutated to determine their role on enzyme activity. We found that Y211A, R212A, R212K and Q214A decreased integration activity, while disintegration and 3'-processing were not significantly affected. By contrast H208A was completely inactive in all the assays. The core domain of Mo-MLV integrase was modeled and the flexibility of the region 207-216 was analyzed. Substitutions with low integration activity showed a lower flexibility than wild type integrase. We propose that the peptide region 207-216 is a flexible loop and that H208, Y211, R212 and Q214 of this loop are involved in the correct assembly of the DNA-integrase complex during integration. PMID- 20026029 TI - The polypeptide Syn67 interacts physically with human holocarboxylase synthetase, but is not a target for biotinylation. AB - Holocarboxylase synthetase (HCS) catalyzes the binding of biotin to lysines in carboxylases and histones in two steps. First, HCS catalyzes the synthesis of biotinyl-5'-AMP; second, the biotinyl moiety is ligated to lysine residues. It has been proposed that step two is fairly promiscuous, and that protein biotinylation may occur in the absence of HCS as long as sufficient exogenous biotinyl-5'-AMP is provided. Here, we identified a novel polypeptide (Syn67) with a basic patch of lysines and arginines. Yeast-two-hybrid assays and limited proteolysis assays revealed that both N- and C-termini of HCS interact with Syn67. A potential target lysine in Syn67 was biotinylated by HCS only after arginine-to-glycine substitutions in Syn67 produced a histone-like peptide. We identified a Syn67 docking site near the active pocket of HCS by in silico modeling and site-directed mutagenesis. Biotinylation of proteins by HCS is more specific than previously assumed. PMID- 20026030 TI - Differential effects of natural and synthetic vitamin E on gene transcription in murine T lymphocytes. AB - Mice were supplemented with low and high doses of natural and synthetic vitamin E, T cells from the spleen isolated and stimulated with plate-bound anti-CD3 and soluble anti-CD28, and gene expression changes assessed by gene array experiments. The data obtained indicate significant qualitative and quantitative differences between the two vitamin forms in regulating gene expression in response to T-cell stimulation. Marker genes have been found whose expression can be considered significant in establishing the level of, and response to vitamin E for both natural and synthetic vitamin E supplementation; unique markers for synthetic vitamin E supplementation and unique markers for natural vitamin E supplementation have been identified. PMID- 20026031 TI - Pyruvate kinase in pig liver mitochondria. AB - The existence of the pyruvate kinase (PK) in pig liver mitochondria was shown by monitoring photometrically the PK reaction in solubilised mitochondria with either phosphoenolpyruvate (PEP) or ADP used as a substrate. In distinction with the cytosolic isoenzyme, the mitochondrial PK showed a sigmoidal dependence on either PEP or ADP concentrations. The occurrence of the mitochondrial PK was confirmed by immunological analysis. Titration with digitonin showed that mPK is restricted to the matrix. PEP addition to mitochondria resulted in reduction of the intramitochondrial NAD(P)+ inhibited by either the non-penetrant thiol reagent mersalyl or by arsenite, an inhibitor of the pyruvate dehydrogenase complex. Citrate/oxaloacetate appearance outside mitochondria also occurred as result of PEP addition to PLM. Taken together these findings support a role for PEP itself in triggering fatty acid synthesis via its mitochondrial metabolism. PMID- 20026033 TI - Are immunoglobulin G4-positive multiorgan lymphoproliferative syndrome and autoimmune pancreatitis different manifestations of a common clinicopathologic entity? PMID- 20026034 TI - Gut microbiota control gut permeability through GLP-2. PMID- 20026036 TI - Antidepressants for irritable bowel syndrome: reappraisal for the nondepressed. PMID- 20026038 TI - Failure of interferon to prevent disease progression and liver cancer in hepatitis C virus infection: proof of absence or absence of proof? PMID- 20026039 TI - Synergistic effect of cAMP and palmitate in promoting altered mitochondrial function and cell death in HepG2 cells. AB - Saturated free fatty acids (FFAs), e.g. palmitate, have long been shown to induce toxicity and cell death in various types of cells. In this study, we demonstrate that cAMP synergistically amplifies the effect of palmitate on the induction of cell death in human hepatocellular carcinoma cell line, HepG2 cells. Elevation of cAMP level in palmitate-treated cells led to enhanced mitochondrial fragmentation, mitochondrial reactive oxygen species (ROS) generation and mitochondrial biogenesis. Mitochondrial fragmentation precedes mitochondrial ROS generation and mitochondrial biogenesis, and may contribute to mitochondrial ROS overproduction and subsequent mitochondrial biogenesis. Fragmentation of mitochondria also facilitated the release of cytotoxic mitochondrial proteins, such as Smac, from the mitochondria and subsequent activation of caspases. However, cell death induced by palmitate and cAMP was caspase-independent and mainly necrotic. PMID- 20026040 TI - Electron transfer in the complex of membrane-bound human cytochrome P450 3A4 with the flavin domain of P450BM-3: the effect of oligomerization of the heme protein and intermittent modulation of the spin equilibrium. AB - We studied the kinetics of NADPH-dependent reduction of human CYP3A4 incorporated into Nanodiscs (CYP3A4-ND) and proteoliposomes in order to probe the effect of P450 oligomerization on its reduction. The flavin domain of cytochrome P450-BM3 (BMR) was used as a model electron donor partner. Unlike CYP3A4 oligomers, where only 50% of the enzyme was shown to be reducible by BMR, CYP3A4-ND could be reduced almost completely. High reducibility was also observed in proteoliposomes with a high lipid-to-protein ratio (L/P=910), where the oligomerization equilibrium is displaced towards monomers. In contrast, the reducibililty in proteoliposomes with L/P=76 did not exceed 55+/-6%. The effect of the surface density of CYP3A4 in proteoliposomes on the oligomerization equilibrium was confirmed with a FRET-based assay employing a cysteine-depleted mutant labeled on Cys-468 with BODIPY iodoacetamide. These results confirm a pivotal role of CYP3A4 oligomerization in its functional heterogeneity. Furthermore, the investigation of the initial phase of the kinetics of CYP3A4 reduction showed that the addition of NADPH causes a rapid low-to-high-spin transition in the CYP3A4-BMR complex, which is followed by a partial slower reversal. This observation reveals a mechanism whereby the CYP3A4 spin equilibrium is modulated by the redox state of the bound flavoprotein. PMID- 20026041 TI - Mutagenesis of the Sauromatum guttatum alternative oxidase reveals features important for oxygen binding and catalysis. AB - The alternative oxidase (AOX) is a non-protonmotive ubiquinol oxidase that is found in mitochondria of all higher plants studied to date. To investigate the role of highly conserved amino acid residues in catalysis we have expressed site directed mutants of Cys-172, Thr-179, Trp-206, Tyr-253, and Tyr-299 in AOX in the yeast Schizosaccharomyces pombe. Assessment of AOX activity in isolated yeast mitochondria reveals that mutagenesis of Trp-206 to phenylalanine or tyrosine abolishes activity, in contrast to that observed with either Tyr-253 or 299 both mutants of which retained activity. None of the mutants exhibited sensitivity to Q-like inhibitors that differed significantly from the wild type AOX. Interestingly, however, mutagenesis of Thr-179 or Cys-172 (a residue implicated in AOX regulation by alpha-keto acids) to alanine not only resulted in a decrease of maximum AOX activity but also caused a significant increase in the enzyme's affinity for oxygen (4- and 2-fold, respectively). These results provide important new insights in the mechanism of AOX catalysis and regulation by pyruvate. PMID- 20026042 TI - Intracranial electrode implantation produces regional neuroinflammation and memory deficits in rats. AB - Deep brain stimulation (DBS) is an established treatment for advanced Parkinson's disease (PD). The procedure entails intracranial implantation of an electrode in a specific brain structure followed by chronic stimulation. Although the beneficial effects of DBS on motor symptoms in PD are well known, it is often accompanied by cognitive impairments, the origin of which is not fully understood. To explore the possible contribution of the surgical procedure itself, we studied the effect of electrode implantation in the subthalamic nucleus (STN) on regional neuroinflammation and memory function in rats implanted bilaterally with stainless steel electrodes. Age-matched sham and intact rats were used as controls. Brains were removed 1 or 8 weeks post-implantation and processed for in vitro autoradiography with [(3)H]PK11195, an established marker of microglial activation. Memory function was assessed by the novel object recognition test (ORT) before surgery and 2 and 8 weeks after surgery. Electrode implantation produced region-dependent changes in ligand binding density in the implanted brains at 1 as well as 8 weeks post-implantation. Cortical regions showed more intense and widespread neuroinflammation than striatal or thalamic structures. Furthermore, implanted animals showed deficits in ORT performance 2 and 8 weeks post-implantation. Thus, electrode implantation resulted in a widespread and persistent neuroinflammation and sustained memory impairment. These results suggest that the insertion and continued presence of electrodes in the brain, even without stimulation, may lead to inflammation-mediated cognitive deficits in susceptible individuals, as observed in patients treated with DBS. PMID- 20026043 TI - Human striatal neuroblasts develop and build a striatal-like structure into the brain of Huntington's disease patients after transplantation. AB - Rebuilding brain structure and neural circuitries by transplantation of fetal tissue is a strategy to repair the damaged nervous system and is currently being investigated using striatal primordium in Huntington's disease (HD) patients. Four HD patients underwent bilateral transplantation with human fetal striatal tissues (9-12 week gestation). Small blocks of whole ganglionic eminencies were processed to obtain cell suspension and then stereotactically grafted in the caudate head and in the putamen. Follow-up period ranged between 18 and 34 months (mean, 24.7 months). Surgery was uneventful. Starting from the fourth month after grafting, neo-generation of metabolically active tissue with striatal-like MRI features was observed in 6 out of 8 grafts. The increase in D2 receptor binding suggested striatal differentiation of the neo-generated tissue in 3 patients. New tissue, connecting the developing grafts with the frontal cortex and, in one case, with the ventral striatum, was also observed. The new tissue growth halted after the ninth month post transplantation. All patients showed stabilization or improvement in some neurological indices. No clinical and imaging signs, suggestive of graft uncontrolled growth, were seen. This study provides the first evidence in humans that neuroblasts of a striatal primordium can develop and move into the brain after neurotransplantation. Primordium development resulted in the building of a new structure with the same imaging features as the corresponding mature structure, combined with short- and long-distance targeted migration of neuroblasts. The results of this study support both the reconstructive potential of fetal tissue and the remarkably retained plasticity of adult brain. Further studies are necessary to assess the clinical efficacy of the human fetal striatal transplantation. PMID- 20026044 TI - NBD-cholesterol probes to track cholesterol distribution in model membranes. AB - A series of cholesterol (Chol) probes with NBD and Dansyl fluorophores attached to the 3-hydroxyl position via carbamate linkers has been designed and synthesized and their ability to mimic the behavior of natural cholesterol in bilayer membranes has been examined. Fluorescence spectroscopy data indicate that the NBD-labeled lipids are located in the polar headgroup region of the bilayer with their position varying with the method of fluorophore attachment and the linker length. The partitioning of the Chol probes between liquid-ordered (L(o)) and liquid-disordered (L(o)) phases in supported bilayers prepared from ternary lipid mixtures of DOPC, Chol and either egg sphingomyelin or DPPC was examined by fluorescence microscopy. The carbamate-linked NBD-Chols show a stronger preference for partitioning into L(o) domains than does a structurally similar probe with an ester linkage, indicating the importance of careful optimization of probe and linker to provide the best Chol mimic. Comparison of the partitioning of NBD probes to literature data for native Chol indicates that the probes reproduce well the modest enrichment of Chol in L(o) domains as well as the ceramide-induced displacement of Chol. One NBD probe was used to follow the dynamic redistribution of Chol in phase separated membranes in response to in situ ceramide generation. This provides the first direct optical visualization of Chol redistribution during enzymatic ceramide generation and allows the assignment of new bilayer regions that exclude dye and have high lateral adhesion to ceramide-rich regions. PMID- 20026045 TI - Ceramide: from lateral segregation to mechanical stress. AB - Ceramide is a sphingolipid present in eukaryotic cells that laterally segregates into solid domains in model lipid membranes. Imaging has provided a wealth of structural information useful to understand some of the physical properties of these domains. In biological membranes, ceramide is formed on one of the membrane leaflets by enzymatic cleavage of sphyngomyelin. Ceramide, with a smaller head size than its parent compound sphyngomyelin, induces an asymmetric membrane tension and segregates into highly ordered domains that have a much high shear viscosity than that of the surrounding lipids. These physical properties, together with the rapid transmembrane flip-flop of the locally produced ceramide, trigger a sequence of membrane perturbations that could explain the molecular mechanism by which ceramide mediates different cell responses. In this review we will try to establish a connection between the physical membrane transformations in model systems known to occur upon ceramide formation and some physiologically relevant process in which ceramide is known to participate. PMID- 20026046 TI - Selective localization of phosphatidylcholine-derived signaling in detergent resistant membranes from synaptic endings. AB - Detergent-resistant membranes (DRMs) are a class of specialized microdomains that compartmentalize several signal transduction processes. In this work, DRMs were isolated from cerebral cortex synaptic endings (Syn) on the basis of their relative insolubility in cold Triton X-100 (1%). The lipid composition and marker protein content were analyzed in DRMs obtained from adult and aged animals. Both DRM preparations were enriched in Caveolin, Flotillin-1 and c-Src and also presented significantly higher sphingomyelin (SM) and cholesterol content than purified Syn. Total phospholipid-fatty acid composition presented an increase in 16:0 (35%), and a decrease in 20:4n-6 (67%) and 22:6n-3 (68%) content in DRM from adults when compared to entire synaptic endings. A more dramatic decrease was observed in the 20:4n-6 and 22:6n-3 content in DRMs from aged animals (80%) with respect to the results found in adults. The coexistence of phosphatidylcholine specific-phospholipase C (PC-PLC) and phospholipase D (PLD) in Syn was previously reported. The presence of these signaling pathways was also investigated in DRMs isolated from adult and aged rats. Both PC-PLC and PLD pathways generate the lipid messenger diacylglycerol (DAG) by catalyzing PC hydrolysis. PC-PLC and PLD1 localization were increased in the DRM fraction. The increase in DAG generation (60%) in the presence of ethanol, confirmed that PC-PLC was also activated when compartmentalized in DRMs. Conversely, PLD2 was excluded from the DRM fraction. Our results show an age-related differential fatty acid composition and a selective localization of PC-derived signaling in synaptic DRMs obtained from adult and aged rats. PMID- 20026047 TI - Screening-based discovery of drug-like O-GlcNAcase inhibitor scaffolds. AB - O-GlcNAcylation is an essential posttranslational modification in metazoa. Modulation of O-GlcNAc levels with small molecule inhibitors of O-GlcNAc hydrolase (OGA) is a useful strategy to probe the role of this modification in a range of cellular processes. Here we report the discovery of novel, low molecular weight and drug-like O-GlcNAcase inhibitor scaffolds by high-throughput screening. Kinetic and X-ray crystallographic analyses of the binding modes with human/bacterial O-GlcNAcases identify some of these as competitive inhibitors. Comparative kinetic experiments with the mechanistically related human lysosomal hexosaminidases reveal that three of the inhibitor scaffolds show selectivity towards human OGA. These scaffolds provide attractive starting points for the development of non-carbohydrate, drug-like OGA inhibitors. PMID- 20026048 TI - Curcumin inhibits hepatitis C virus replication via suppressing the Akt-SREBP-1 pathway. AB - A polyphenolic compound from the curry spice turmeric, curcumin, is known to show anti-viral activity against the influenza virus, adenovirus, coxsackievirus, and the human immunodeficiency virus. However, it remains to be determined whether curcumin can inhibit the replication of hepatitis C virus (HCV). In this study, we showed that curcumin decreases HCV gene expression via suppression of the Akt SREBP-1 activation, not by NF-kappaB pathway. The combination of curcumin and IFNalpha exerted profound inhibitory effects on HCV replication. Collectively, our results indicate that curcumin can suppress HCV replication in vitro and may be potentially useful as novel anti-HCV reagents. PMID- 20026049 TI - PCSK9-deficient mice exhibit impaired glucose tolerance and pancreatic islet abnormalities. AB - Proprotein convertase subtilisin/kexin type 9 (PCSK9), a liver-secreted plasma enzyme, restricts hepatic uptake of low-density lipoprotein (LDL) cholesterol by promoting the degradation of LDL receptors (LDLR). PCSK9 and LDLR are also expressed in insulin-producing pancreatic islet beta cells, possibly affecting the function of these cells. Here we show that, compared to control mice, PCSK9 null male mice over 4 months of age carried more LDLR and less insulin in their pancreas; they were hypoinsulinemic, hyperglycemic and glucose-intolerant; their islets exhibited signs of malformation, apoptosis and inflammation. Collectively, these observations suggest that PCSK9 may be necessary for the normal function of pancreatic islets. PMID- 20026051 TI - Molecular mechanism of bundle formation by the bacterial actin ParM. AB - The actin homolog ParM plays a microtubule-like role in segregating DNA prior to bacterial cell division. Fluorescence and cryo-electron microscopy have shown that ParM forms filament bundles between separating DNA plasmids in vivo. Given the lack of ParM bundling proteins it remains unknown how ParM bundles form at the molecular level. Here we show using time-lapse TIRF microscopy, under in vitro molecular crowding conditions, that ParM-bundle formation consists of two distinct phases. At the onset of polymerization bundle thickness and shape are determined in the form of nuclei of short helically disordered filaments arranged in a liquid-like lattice. These nuclei then undergo an elongation phase whereby they rapidly increase in length. At steady state, ParM bundles fuse into one single large aggregate. This behavior had been predicted by theory but has not been observed for any other cytomotive biopolymer, including F-actin. We employed electron micrographs of ParM rafts, which are 2-D analogs of 3-D bundles, to identify the main molecular interfilament contacts within these suprastructures. The interface between filaments is similar for both parallel and anti-parallel orientations and the distribution of filament polarity is random within a bundle. We suggest that the interfilament interactions are not due to the interactions of specific residues but rather to long-range, counter ion mediated, electrostatic attractive forces. A randomly oriented bundle ensures that the assembly is rigid and that DNA may be captured with equal efficiency at both ends of the bundle via the ParR binding protein. PMID- 20026050 TI - Residue Glu83 plays a major role in negatively regulating alpha-synuclein amyloid formation. AB - Alpha-synuclein (alpha-syn) amyloid filaments are the major ultrastructural component of pathological inclusions that define several neurodegenerative disorders, including Parkinson disease and other disorders that are collectively termed synucleinopathies. Since the aggregation of alpha-syn is associated with the etiology of these diseases, defining the molecular elements that influence this process may have important therapeutics implication. The deletions of major portions of the hydrophobic region of alpha-syn (Delta74-79 and Delta71-82) impair the ability to form amyloid. However, mutating residue E83 to an A restored the ability of these proteins to form amyloid. Additionally supporting an inhibitory role of residue E83 on amyloid formation, mutating this residue to an A enhanced amyloid formation in the presence of small molecule inhibitors, such as dopamine and EGCG. Our data, therefore, suggest that the presence and placement of the highly charged E83 residue plays a significant inhibitory role in alpha-syn amyloid formation and these findings provide important insights in the planning of therapeutic agents that may be capable of preventing alpha-syn amyloid formation. PMID- 20026053 TI - WITHDRAWN: Relationships between 8q24 and 17q risk loci and sporadic or latent prostate cancer and the impacts of these loci on the clinicopathologic characteristics of prostate cancer. AB - This article has been withdrawn at the request of the editor. The Publisher apologizes for any inconvenience this may cause. The full Elsevier Policy on Article Withdrawal can be found at http://www.elsevier.com/locate/withdrawalpolicy. PMID- 20026052 TI - Decorin is processed by three isoforms of bone morphogenetic protein-1 (BMP1). AB - The secreted small proteoglycan, decorin, modulates collagen fibril formation as well as the bioactivity of various members of the transforming growth factor-beta (TGFbeta) superfamily. Indeed, recombinant prodecorin has been used in several gene therapy experiments to inhibit unwanted fibrosis in model diseases of the kidney, heart, and other tissues although the status of the propeptide within the target tissues is unknown. Currently the protease that removes the highly conserved propeptide from decorin is unproven. Using a variety of approaches, we show that three isoforms of the Tolloid-related bone morphogenetic protein-1 (BMP1) can effectively remove the propeptide from human prodecorin resulting in the well-established mature proteoglycan. Classic BMP1, the full-length gene transcript mTLD (BMP1-3), and BMP1-5 (isoform lacking the CUB3 domain thought to be important for efficient type I collagen C-propeptidase activity) all removed the analogous propeptides from both recombinant human prodecorin and murine probiglycan. Furthermore, the timed removal of the propeptide was found to not be necessary for the addition of decorin's single glycosaminoglycan chain. Decorin therefore joins the growing list of matrix and bioactive molecules processed/activated by the BMP1/Tolloid family. Since the third member of the Class I small leucine-rich proteooglycan (SLRP) superfamily, asporin, also contains a similar cleavage motif at the appropriate location, we propose that the removal of these propeptides by members of the BMP1 family is an additional characteristic of Class I SLRP. PMID- 20026054 TI - RON-expressing MCF-10A breast epithelial cells exhibit alterations of hyaluronan expression, promoting RON-mediated early adhesion events. AB - The receptor tyrosine kinase known as RON appears to play a role in the progression of human carcinomas, and is associated with a poor patient prognosis. Our current study demonstrates that RON expression in MCF-10A breast epithelial cells lead to an alteration of cell-surface hyaluronan compared to the parental cells. We found that hyaluronan was important for initial cell attachment to poly d-lysine-coated coverslips, but did not contribute to the process of cell spreading. Previous data implied that the Src kinase was important for spreading but not the initial attachment of 10A cells, and here we demonstrate Src activation was also not necessary for hyaluronan production in these cells. PMID- 20026055 TI - Apoptosis repressor with caspase recruitment domain is dramatically reduced in cardiac, skeletal, and vascular smooth muscle during hypertension. AB - Apoptosis repressor with caspase recruitment domain (ARC) is a unique anti apoptotic protein with a distinct tissue distribution. In addition, unlike most anti-apoptotic proteins which act on one pathway, ARC can inhibit apoptosis mediated by both the death-receptor and mitochondrial signaling pathways. In this study, we confirm previous reports showing high levels of ARC protein in rat heart and skeletal muscle, but demonstrate for the first time that ARC is also expressed in rat aorta. Immunoblot analysis on endothelium-denuded aorta as well as immunohistochemical analysis on intact aorta demonstrated that ARC was highly expressed in smooth muscle. Immunoblot analysis also found that ARC protein was severely downregulated in skeletal muscle (-82%; P<0.001), heart (-80%; P<0.001), and aorta (-71%; P<0.001) of spontaneously hypertensive rats (SHR) compared to normotensive Wistar-Kyoto (WKY) rats. Decreased ARC levels were also confirmed in tissues of hypertensive animals by immunohistochemical analysis. Collectively, this data suggests that ARC protein is expressed in vascular smooth muscle and is significantly reduced in several target tissues during hypertension. PMID- 20026056 TI - IL-6 and IFN-alpha from dsRNA-stimulated dendritic cells control expansion of regulatory T cells. AB - Foxp3(+)CD4(+) regulatory T cells (Treg) control not only autoimmunity but also the effective immune response against RNA virus infections, which produces virus derived double-stranded RNA (dsRNA). To induce effective anti-viral immunity, it is a key issue to learn how Treg respond to dsRNA in vitro and in vivo. We here showed that synthetic dsRNA, polyI:C, caused peripheral expansion of functional Treg in a TICAM-1- and IL-6-dependent manner in vivo. PolyI:C did not expand Treg directly, but promoted the expansion of naturally occurring Treg indirectly through IL-6 produced from dendritic cells (DCs). In addition, the expansion of Treg by IL-6 was inhibited by IFN-alpha from polyI:C-stimulated DCs. These data suggest that the balance of IL-6 and IFN-alpha in the region of RNA virus infection may determine the number of peripheral Treg, which affects the effective immune responses against viruses. PMID- 20026057 TI - Toxicity of a serotonin-derived neuromelanin. AB - Postoperative Cognitive Dysfunction (POCD) is associated with increased mortality in the elderly and may occur from lipid peroxidation in aging. We previously showed that sevoflurane sequesters acrolein, which promotes the formation of a novel species of a putative neuromelanin. The current study examined the properties of this serotonin-derived melanoid (SDM). The interaction of SDM with unilamellar vesicles (ULVs) was examined using lipid membrane probes. Vesicle disruption was investigated by leakage of dye from calcein-loaded ULVs. We observed that SDM decreased diphenyl-hexatriene fluorescence anisotropy and increased the temperature-dependent change in anisotropy. SDM changed the absorbance of merocyanin-bound ULVs. SDM increased detergent-mediated calcein leakage. SDM structure was dramatically altered upon interaction with ULVs. We also observed that SDM enhanced detergent-mediated leakage of loaded ULVs, suggesting that SDM may be neurotoxic. We propose that inhalational agents, which sequester acrolein, may promote the production of certain species of neuromelanin that depletes local serotonin and enhances neuronal vulnerability. PMID- 20026058 TI - Impaired methylation as a novel mechanism for proteasome suppression in liver cells. AB - The proteasome is a multi-catalytic protein degradation enzyme that is regulated by ethanol-induced oxidative stress; such suppression is attributed to CYP2E1 generated metabolites. However, under certain conditions, it appears that in addition to oxidative stress, other mechanisms are also involved in proteasome regulation. This study investigated whether impaired protein methylation that occurs during exposure of liver cells to ethanol, may contribute to suppression of proteasome activity. We measured the chymotrypsin-like proteasome activity in Huh7CYP cells, hepatocytes, liver cytosols and nuclear extracts or purified 20S proteasome under conditions that maintain or prevent protein methylation. Reduction of proteasome activity of hepatoma cell and hepatocytes by ethanol or tubercidin was prevented by simultaneous treatment with S-adenosylmethionine (SAM). Moreover, the tubercidin-induced decline in proteasome activity occurred in both nuclear and cytosolic fractions. In vitro exposure of cell cytosolic fractions or highly purified 20S proteasome to low SAM:S-adenosylhomocysteine (SAH) ratios in the buffer also suppressed proteasome function, indicating that one or more methyltransferase(s) may be associated with proteasomal subunits. Immunoblotting a purified 20S rabbit red cell proteasome preparation using methyl lysine-specific antibodies revealed a 25kDa proteasome subunit that showed positive reactivity with anti-methyl lysine. This reactivity was modified when 20S proteasome was exposed to differential SAM:SAH ratios. We conclude that impaired methylation of proteasome subunits suppressed proteasome activity in liver cells indicating an additional, yet novel mechanism of proteasome activity regulation by ethanol. PMID- 20026059 TI - U6 promoter-driven siRNA injection has nonspecific effects in zebrafish. AB - RNA interference (RNAi) is a posttranscriptional gene silencing mechanism triggered by double-stranded RNA (dsRNA), which causes degradation of homologous mRNAs. RNAi has been observed in a wide range of eukaryotes, including fungi, plants and animals. In vertebrates, long dsRNA activates the interferon response and yields nonspecific degradation of mRNA. In contrast, small interference RNA (siRNA) duplexes with a length of 21-23 nucleotides trigger specific gene silencing and thus are widely used in gene function studies. The use of siRNA for gene silencing in zebrafish has rarely been reported. In this report, we studied mammalian U6 promoter-driven siRNA-mediated RNA interference in zebrafish. The well characterized genes Myf5, Dlg3 and Nacre were selected as targets. Two to four target siRNAs were synthesized with incorporation of the U6 promoter. Constructs were introduced into early zebrafish embryos through microinjection, followed by in situ hybridization and embryonic development was monitored to determine whether U6 promoter-driven siRNAs could efficiently suppress specific gene expression. We showed that these siRNAs could partially suppress endogenous gene expression and that the siRNA efficiency varied at different targeted positions. However, the U6 promoter-driven siRNAs may also have induced nonspecific gene suppression (off-target effects). It appears that, despite the findings of previous reports, the current methodology of siRNA interference is not practical for studying gene function during early zebrafish development. PMID- 20026060 TI - Blockade of the ERK or PI3K-Akt signaling pathway enhances the cytotoxicity of histone deacetylase inhibitors in tumor cells resistant to gefitinib or imatinib. AB - Deregulated activation of protein tyrosine kinases, such as the epidermal growth factor receptor (EGFR) and Abl, is associated with human cancers including non small cell lung cancer (NSCLC) and chronic myeloid leukemia (CML). Although inhibitors of such activated kinases have proved to be of therapeutic benefit in individuals with NSCLC or CML, some patients manifest intrinsic or acquired resistance to these drugs. We now show that, whereas blockade of either the extracellular signal-regulated kinase (ERK) pathway or the phosphatidylinositol 3 kinase (PI3K)-Akt pathway alone induced only a low level of cell death, it markedly sensitized NSCLC or CML cells to the induction of apoptosis by histone deacetylase (HDAC) inhibitors. Such enhanced cell death induced by the respective drug combinations was apparent even in NSCLC or CML cells exhibiting resistance to EGFR or Abl tyrosine kinase inhibitors, respectively. Co-administration of a cytostatic signaling pathway inhibitor may contribute to the development of safer anticancer strategies by lowering the required dose of cytotoxic HDAC inhibitors for a variety of cancers. PMID- 20026061 TI - The potential of the cruciform structure formation as an important factor influencing p53 sequence-specific binding to natural DNA targets. AB - p53 is one of the most important tumor suppressors which responds to DNA damage by binding to DNA and regulating the transcription of genes involved in cell cycle arrest, apoptosis, or senescence. As it was shown previously, p53 binding to DNA is strongly influenced by DNA topology. DNA supercoiling is fundamentally important for a wide range of biological processes including DNA transcription, replication, recombination, control of gene expression and genome organization. In this study, we investigated the cruciform structures formation of various inverted repeats in p53-responsive sequences from p21, RGC, mdm2 and GADD45 promoters under negative superhelical stress, and analyzed the effects of these DNA topology changes on p53-DNA binding. We demonstrated using three different methods (gel retardation analyses, ELISA and magnetic immunoprecipitation assay) that the p53 protein binds preferentially to negatively supercoiled plasmid DNAs with p53-responsive sequence presented as a cruciform structure. Not only the appearance of the cruciform structures within naked supercoiled DNA, but also the potential of the binding sites for adopting the non-B structures can contribute to a more favorable p53-DNA complex. PMID- 20026062 TI - Substrate overload: Glucose oxidation in human myotubes conquers palmitate oxidation through anaplerosis. AB - To date, two cardinal principles govern oxidation of glucose and fatty acids in skeletal muscle; exogenous fatty acid reduces glucose oxidation and glucose reduces fatty acid oxidation. Both glucose and palmitate (PA) oxidation was increased by increasing their concentration and inhibited by increasing concentrations of the other in human myotubes established from healthy, lean subjects exposed to acute stepwise increases in glucose and PA levels. At high substrate levels; PA oxidation was reduced while release of acid soluble metabolites was increased and, both glucose oxidation and release of citrate was increased which could be abolished by phenylacetic acid (inhibitor of pyruvate carboxylase (PC)). The present data challenges above preconceptions. Although they operate at low-moderate substrate levels additional two principles determine substrate oxidation at higher substrate concentrations; first, anaplerosis of the tricarboxylic cycle through PC promoting complete and incomplete glucose oxidation; second, inhibition of complete PA oxidation with increasing incomplete PA oxidation mediated by high glucose and PA levels, respectively. PMID- 20026063 TI - Mechanism of macrophage activation induced by beta-glucan produced from Paenibacillus polymyxa JB115. AB - Beta-glucans are heterogeneous groups of glucose polymers found in the cell walls of fungi, plants and some bacteria. Our previous report showed that a novel beta 1,3/1,6-glucan produced from Paenibacillus (P.) polymyxa JB115 can induce nitric oxide (NO) production in RAW264.7 cells. In the present study, the beta-glucan significantly increased luciferase activity in cells transfected with NFkappaB or AP1, but not STAT1, reporter vector DNA, which contain their binding promoter site. All specific NFkappaB and MAPKs pathway inhibitors (pyrrolidine dithiocarbamate, AG490, U0126, SB203580 and SP600125) remarkably attenuated NO production induced by the beta-glucan. Furthermore, Western blot analysis revealed that the stimulation of Raw264.7 cells by beta-glucan induced phosphorylation of IkappaB and the consequent translocation of NFkappaB into the nucleus. Meanwhile, phosphorylation of ERK1/2, JNK/SAPK and p38 MAPKs in cytoplasm were also confirmed. All these results indicated that beta-glucan from P. polymyxa JB115 activates macrophages through MAPKs and NFkappaB signaling pathway. PMID- 20026064 TI - Prostaglandin E2 prevents Helicobacter-induced gastric preneoplasia and facilitates persistent infection in a mouse model. AB - BACKGROUND & AIMS: Persistent infection with the human pathogen Helicobacter pylori increases the risk of gastric cancer. In this study, we investigated the role of cyclooxygenase-2 (COX-2) and its main product, prostaglandin E(2) (PGE(2)), in the development of Helicobacter-induced gastritis and gastric cancer precursor lesions. METHODS: We utilized mouse models of Helicobacter-induced gastric preneoplasia and vaccine-induced protection to study the effects of COX-2 inhibition and PGE(2) treatment on the induction of Helicobacter-specific immune responses and gastric premalignant immunopathology. RESULTS: COX-2 and PGE(2) are up-regulated upon Helicobacter infection in cultured epithelial cells and in the gastric mucosa of infected mice. Inhibition of COX-2 activity with celecoxib significantly accelerated early preneoplasia; conversely, systemic administration of synthetic PGE(2) prevented development of premalignant pathology and completely reversed preexisting lesions by suppressing interferon-gamma production in the infected stomachs. The protective effect of PGE(2) was accompanied by increased Helicobacter colonization in all models. All in vivo effects were attributed to immunosuppressive effects of PGE(2) on CD4(+) T-helper 1 cells, which fail to migrate, proliferate, and secrete cytokines when exposed to PGE(2) in vitro and in vivo. T-cell inhibition was found to be due to silencing of interleukin-2 gene transcription, and could be overcome by supplementation with recombinant interleukin-2 in vitro and in vivo. CONCLUSIONS: COX-2-dependent production of PGE(2) has an important immunomodulatory role during Helicobacter infection, preventing excessive local immune responses and the associated immunopathology by inhibiting the effector functions of pathogenic T-helper 1 cells. PMID- 20026065 TI - Hip fracture risk in patients with a diagnosis of pernicious anemia. AB - BACKGROUND & AIMS: Pernicious anemia (PA) is characterized by vitamin B-12 deficiency and achlorhydria, both of which have a detrimental effect on bone strength. The principle aim of this study was to determine the risk of hip fracture in patients with PA. METHODS: This is a retrospective cohort study using the General Practice Research Database (GPRD) from the United Kingdom. GPRD data from May 1987 until April 2002 were utilized to identify patients between 40 and 90 years of age at the time of GPRD enrollment. The exposed group contained patients with a diagnosis of PA being treated with vitamin B-12 therapy. We matched each patient having a diagnosis of PA with 4 randomly selected non-PA patients with respect to age (+/-1 year) and sex. Cox regression analysis was used to determine the hazard ratio (HR) for hip fracture associated with PA. RESULTS: Nine thousand five hundred six patients with a diagnosis of PA receiving vitamin B-12 injection therapy were identified and compared to 38,024 controls. Patients with PA had a greater risk of hip fracture than the controls (HR = 1.74; 95% CI: 1.45-2.08). The increase in hip fracture risk was even more pronounced among those patients newly diagnosed with PA during GPRD follow-up (HR = 2.63; 95% CI: 2.03-3.41). CONCLUSIONS: Patients with a diagnosis of PA have an elevated risk of hip fracture. The increased hip fracture risk was persistent even years after vitamin B-12 therapy. Chronic achlorhydria could be the mechanism contributing to the persistently elevated hip fracture risk. PMID- 20026066 TI - Pancreatic duct glands are distinct ductal compartments that react to chronic injury and mediate Shh-induced metaplasia. AB - BACKGROUND & AIMS: Pancreatic intraepithelial neoplasia (PanIN) are pancreatic cancer precursor lesions of unclear origin and significance. PanIN aberrantly express sonic hedgehog (Shh), an initiator of pancreatic cancer, and gastrointestinal mucins. A majority of PanIN are thought to arise from ducts. We identified a novel ductal compartment that is gathered in gland-like outpouches (pancreatic duct glands [PDG]) of major ducts and characterized its role in injury and metaplasia. METHODS: The ductal system was analyzed in normal pancreata and chronic pancreatitis in humans and mice. Anatomy was assessed by serial hematoxylin and eosin sections and scanning electron microscopy of corrosion casts. Expression of mucins and developmental genes and proliferation were assessed by immunohistochemistry or real-time quantitative polymerase chain reaction. Effects of Shh on ductal cells were investigated by exposure to Shh in vitro and transgenic misexpression in vivo. RESULTS: Three-dimensional analysis revealed blind-ending outpouches of ducts in murine and human pancreata. These PDG are morphologically and molecularly distinct from normal ducts; even in normal pancreata they display PanIN and metaplastic features, such as expression of Shh and gastric mucins. They express other developmental genes, such as Pdx-1 and Hes-1. In injury, Shh is up-regulated along with gastric mucins. Expansion of the PDG compartment results in a mucinous metaplasia. Shh promotes this transformation in vitro and in vivo. CONCLUSIONS: PDG are distinct gland-like mucinous compartments with a distinct molecular signature. In response to injury, PDG undergo an Shh-mediated mucinous gastrointestinal metaplasia with PanIN-like features. PDG may provide a link between Shh, mucinous metaplasia, and neoplasia. PMID- 20026067 TI - Isolation of diapause-regulated genes from the flesh fly, Sarcophaga crassipalpis by suppressive subtractive hybridization. AB - Subtractive suppressive hybridization (SSH) was used to characterize the diapause transcriptome of the flesh fly Sarcophaga crassipalpis. Through these efforts, we isolated 97 unique clones which were used as probes in northern hybridization to assess their expression during diapause. Of these, 17 were confirmed to be diapause upregulated and 1 was diapause downregulated, while 12 were shown to be unaffected by diapause in this species. The diapause upregulated genes fall into several broad categories including heat shock proteins, heavy metal responsive genes, neuropeptides, structural genes, regulatory elements, and several genes of unknown function. In combination with other large-scale analyses of gene expression during diapause, this study assists in the characterization of the S. crassipalpis diapause transcriptome, and begins to identify common elements involved in diapause across diverse taxa. PMID- 20026068 TI - Crystal structure of the LasA virulence factor from Pseudomonas aeruginosa: substrate specificity and mechanism of M23 metallopeptidases. AB - Pseudomonas aeruginosa is an opportunist Gram-negative bacterial pathogen responsible for a wide range of infections in immunocompromized individuals and is a leading cause of mortality in cystic fibrosis patients. A number of secreted virulence factors, including various proteolytic enzymes, contribute to the establishment and maintenance of Pseudomonas infection. One such is LasA, an M23 metallopeptidase related to autolytic glycylglycine endopeptidases such as Staphylococcus aureus lysostaphin and LytM, and to DD-endopeptidases involved in entry of bacteriophage to host bacteria. LasA is implicated in a range of processes related to Pseudomonas virulence, including stimulating ectodomain shedding of the cell surface heparan sulphate proteoglycan syndecan-1 and elastin degradation in connective tissue. Here we present crystal structures of active LasA as a complex with tartrate and in the uncomplexed form. While the overall fold resembles that of the other M23 family members, the LasA active site is less constricted and utilizes a different set of metal ligands. The active site of uncomplexed LasA contains a five-coordinate zinc ion with trigonal bipyramidal geometry and two metal-bound water molecules. Using these structures as a starting point, we propose a model for substrate binding by LasA that explains its activity against a wider range of substrates than those used by related lytic enzymes, and offer a catalytic mechanism for M23 metallopeptidases consistent with available structural and mutagenesis data. Our results highlight how LasA is a structurally distinct member of this endopeptidase family, consistent with its activity against a wider range of substrates and with its multiple roles in Pseudomonas virulence. PMID- 20026069 TI - A bacterial transcription terminator with inefficient molecular motor action but with a robust transcription termination function. AB - Molecular motors such as helicases/translocases are capable of translocating along the single-stranded nucleic acids and unwinding DNA or RNA duplex substrates using the energy derived from their ATPase activity. The bacterial transcription terminator, Rho, is a hexameric helicase and releases RNA from the transcription elongation complexes by an unknown mechanism. It has been proposed, but not directly demonstrated, that kinetic energy obtained from its molecular motor action (helicase/translocase activities) is instrumental in dissociating the transcription elongation complex. Here we report a hexameric Rho analogue (Rv1297, M. tb. Rho) from Mycobacterium tuberculosis having poor RNA-dependent ATP hydrolysis and inefficient DNA-RNA unwinding activities. However, compared to Escherichia coli Rho, it exhibited very robust and earlier transcription termination from the elongation complexes of E. coli RNA polymerase. Bicyclomycin, an inhibitor of ATPase as well as RNA release activities of E. coli Rho, inhibited the ATPase activity of M. tb. Rho with comparable efficiency but was not efficient in inhibiting its transcription termination function. Unlike E. coli Rho, M. tb. Rho was capable of releasing RNA in the presence of nonhydrolyzable analogues of ATP quite efficiently. Also, this termination function most likely does not require NusG, an RNA-release facilitator, as this Rho was incapable of binding to NusG either of M. tb. (Rv0639) or E. coli. These results strongly suggest that the ATPase activity of M. tb. Rho is uncoupled from its transcription termination function and this function may not be dependent on its helicase/translocase activity. PMID- 20026070 TI - Functional analysis of human tRNA isodecoders. AB - tRNA isodecoders share the same anticodon but have differences in their body sequence. An unexpected result from genome sequencing projects is the identification of a large number of tRNA isodecoder genes in mammalian genomes. In the reference human genome, more than 270 isodecoder genes are present among the approximately 450 tRNA genes distributed among 49 isoacceptor families. Whether sequence diversity among isodecoder tRNA genes reflects functional variability is an open question. To address this, we developed a method to quantify the efficiency of tRNA isodecoders in stop-codon suppression in human cell lines. First, a green fluorescent protein (GFP) gene that contains a single UAG stop codon at two distinct locations is introduced. GFP is only produced when a tRNA suppressor containing CUA anticodon is co-transfected with the GFP gene. The suppression efficiency is examined for 31 tRNA isodecoders (all contain CUA anticodon), 21 derived from four isoacceptor families of tRNASer genes, 7 from five families of tRNALeu genes, and 3 from three families of tRNAAla genes. We found that isodecoder tRNAs display a large difference in their suppression efficiency. Among those with above background suppression activity, differences of up to 20-fold were observed. We were able to tune tRNA suppression efficiency by subtly adjusting the tRNA sequence and inter-convert poor suppressors into potent ones. We also demonstrate that isodecoder tRNAs with varying suppression efficiencies have similar stability and exhibit similar levels of aminoacylation in vivo. Our results indicate that naturally occurring tRNA isodecoders can have large functional variations and suggest that some tRNA isodecoders may perform a function distinct from translation. PMID- 20026072 TI - Structural and functional characterization of plant aminoaldehyde dehydrogenase from Pisum sativum with a broad specificity for natural and synthetic aminoaldehydes. AB - Aminoaldehyde dehydrogenases (AMADHs, EC 1.2.1.19) belong to the large aldehyde dehydrogenase (ALDH) superfamily, namely, the ALDH9 family. They oxidize polyamine-derived omega-aminoaldehydes to the corresponding omega-amino acids. Here, we report the first X-ray structures of plant AMADHs: two isoenzymes, PsAMADH1 and PsAMADH2, from Pisum sativum in complex with beta-nicotinamide adenine dinucleotide (NAD(+)) at 2.4 and 2.15 A resolution, respectively. Both recombinant proteins are dimeric and, similarly to other ALDHs, each monomer is composed of an oligomerization domain, a coenzyme binding domain and a catalytic domain. Each subunit binds NAD(+) as a coenzyme, contains a solvent-accessible C terminal peroxisomal targeting signal (type 1) and a cation bound in the cavity close to the NAD(+) binding site. While the NAD(+) binding mode is classical for PsAMADH2, that for PsAMADH1 is unusual among ALDHs. A glycerol molecule occupies the substrate binding site and mimics a bound substrate. Structural analysis and substrate specificity study of both isoenzymes in combination with data published previously on other ALDH9 family members show that the established categorization of such enzymes into distinct groups based on substrate specificity is no more appropriate, because many of them seem capable of oxidizing a large spectrum of aminoaldehyde substrates. PsAMADH1 and PsAMADH2 can oxidize N,N,N-trimethyl-4 aminobutyraldehyde into gamma-butyrobetaine, which is the carnitine precursor in animal cells. This activity highly suggests that in addition to their contribution to the formation of compatible osmolytes such as glycine betaine, beta-alanine betaine and gamma-aminobutyric acid, AMADHs might participate in carnitine biosynthesis in plants. PMID- 20026071 TI - Modulation of polyglutamine conformations and dimer formation by the N-terminus of huntingtin. AB - Polyglutamine expansions within different proteins are associated with nine different neurodegenerative diseases. There is growing interest in understanding the roles of flanking sequences from disease-relevant proteins in the intrinsic conformational and aggregation properties of polyglutamine. We report results from atomistic simulations and circular dichroism experiments that quantify the effect of the N-terminal 17-residue (Nt17) segment of the huntingtin protein on polyglutamine conformations and intermolecular interactions. We show that the Nt17 segment and polyglutamine domains become increasingly disordered as polyglutamine length (N) increases in Nt17-Q(N) constructs. Hydrophobic groups within Nt17 become sequestered in intramolecular interdomain interfaces. We also show that the Nt17 segment suppresses the intrinsic propensity of polyglutamine aggregation. This inhibition arises from the incipient micellar structures adopted by monomeric forms of the peptides with Nt17 segments. The degree of intermolecular association increases with increasing polyglutamine length and is governed mainly by associations between polyglutamine domains. Comparative analysis of intermolecular associations for different polyglutamine-containing constructs leads to clearer interpretations of recently published experimental data. Our results suggest a framework for fibril formation and identify roles for flanking sequences in the modulation of polyglutamine aggregation. PMID- 20026073 TI - Modulation of an active-site cysteine pKa allows PDI to act as a catalyst of both disulfide bond formation and isomerization. AB - Protein disulfide isomerase (PDI) plays a central role in disulfide bond formation in the endoplasmic reticulum. It is implicated both in disulfide bond formation and in disulfide bond reduction and isomerization. To be an efficient catalyst of all three reactions requires complex mechanisms. These include mechanisms to modulate the pK(a) values of the active-site cysteines of PDI. Here, we examined the role of arginine 120 in modulating the pK(a) values of these cysteines. We find that arginine 120 plays a significant role in modulating the pK(a) of the C-terminal active-site cysteine in the a domain of PDI and plays a role in determining the reactivity of the N-terminal active-site cysteine but not via direct modulation of its pK(a). Mutation of arginine 120 and the corresponding residue, arginine 461, in the a' domain severely reduces the ability of PDI to catalyze disulfide bond formation and reduction but enhances the ability to catalyze disulfide bond isomerization due to the formation of more stable PDI-substrate mixed disulfides. These results suggest that the modulation of pK(a) of the C-terminal active cysteine by the movement of the side chain of these arginine residues into the active-site locales has evolved to allow PDI to efficiently catalyze both oxidation and isomerization reactions. PMID- 20026074 TI - The three-dimensional structure of [NiFeSe] hydrogenase from Desulfovibrio vulgaris Hildenborough: a hydrogenase without a bridging ligand in the active site in its oxidised, "as-isolated" state. AB - Hydrogen is a good energy vector, and its production from renewable sources is a requirement for its widespread use. [NiFeSe] hydrogenases (Hases) are attractive candidates for the biological production of hydrogen because they are capable of high production rates even in the presence of moderate amounts of O(2), lessening the requirements for anaerobic conditions. The three-dimensional structure of the [NiFeSe] Hase from Desulfovibrio vulgaris Hildenborough has been determined in its oxidised "as-isolated" form at 2.04-A resolution. Remarkably, this is the first structure of an oxidised Hase of the [NiFe] family that does not contain an oxide bridging ligand at the active site. Instead, an extra sulfur atom is observed binding Ni and Se, leading to a SeCys conformation that shields the NiFe site from contact with oxygen. This structure provides several insights that may explain the fast activation and O(2) tolerance of these enzymes. PMID- 20026075 TI - Structure of the adenylylation domain of E. coli glutamine synthetase adenylyl transferase: evidence for gene duplication and evolution of a new active site. AB - The X-ray structure of the C-terminal fragment, containing residues 449-946, of Escherichia coli glutamine synthetase adenylyl transferase (ATase) has been determined. ATase is part of the cascade that regulates the enzymatic activity of E. coli glutamine synthetase, a key component of the cell's machinery for the uptake of ammonia. It has two enzymatic activities, adenylyl removase (AR) and adenylyl transferase (AT), which are located in distinct catalytic domains that are separated by a regulatory (R) domain. We previously reported the three dimensional structure of the AR domain (residues 1-440). The present structure contains both the R and AT domains. AR and AT share 24% sequence identity and also contain the beta-polymerase motif that is characteristic of many nucleotidylyl transferase enzymes. The structures overlap with an rmsd of 2.4 A when the superhelical R domain is omitted. A model for the complete ATase molecule is proposed, along with some refinements of domain boundaries. A rather more speculative model for the complex of ATase with glutamine synthetase and the nitrogen signal transduction protein PII is also presented. PMID- 20026077 TI - Conversion of Abeta42 into a folded soluble native-like protein using a semi random library of amphipathic helices. AB - The amyloid cascade model hypothesizes that neurotoxic oligomers or aggregates formed by the Alzheimer amyloid peptide (Abeta) cause disease pathology in Alzheimer's disease. Attempted treatment strategies for Alzheimer's disease have involved either inhibiting Abeta oligomerization or aggregation, or dissolving existing aggregates. Blocking such downhill processes, however, has proved daunting. We have used a different approach that targets Abeta before the oligomerization cascade begins. We predicted that an amphipathic helix could convert Abeta into a native-like protein and inhibit initiation of oligomerization and aggregation. This idea was tested with a designed library and genetic screen. We exhaustively screened a library of semi-randomized amphipathic helical sequences, each expressed as a fusion protein with an Abeta42-yellow fluorescent protein sequence serving as a reporter for folding and solubilization. This yielded an amphipathic helix capable of initiating native like folding in Abeta42 and preventing aggregation. This amphipathic helix has direct application to Alzheimer's disease therapy development. PMID- 20026076 TI - Accessory alpha-helix of complexin I can displace VAMP2 locally in the complexin SNARE quaternary complex. AB - The calcium-triggered neurotransmitter release requires three SNARE (soluble N ethylmaleimide-sensitive factor attachment protein receptor) proteins: synaptobrevin 2 (or vesicle-associated membrane protein 2) on the synaptic vesicle and syntaxin 1 and SNAP-25 (synaptosome-associated protein of 25 kDa) at the presynaptic plasma membrane. This minimal fusion machinery is believed to drive fusion of the vesicle to the presynaptic membrane. Complexin, also known as synaphin, is a neuronal cytosolic protein that acts as a major regulator of synaptic vesicle exocytosis. Stimulatory and inhibitory effects of complexin have both been reported, suggesting the duality of its function. To shed light on the molecular basis of the complexin's dual function, we have performed an EPR investigation of the complexin-SNARE quaternary complex. We found that the accessory alpha-helix (amino acids 27-48) by itself has the capacity to replace the C-terminus of the SNARE motif of vesicle-associated membrane protein 2 in the four-helix bundle and makes the SNARE complex weaker when the N-terminal region of complexin I (amino acids 1-26) is removed. However, the accessory alpha-helix remains detached from the SNARE core when the N-terminal region of complexin I is present. Thus, our data show the possibility that the balance between the activities of the accessory alpha-helix and the N-terminal domain might determine the final outcome of the complexin function, either stimulatory or inhibitory. PMID- 20026078 TI - Binding of S-methyl-5'-thioadenosine and S-adenosyl-L-methionine to protein MJ0100 triggers an open-to-closed conformational change in its CBS motif pair. AB - Cystathionine beta-synthase (CBS) domains are small motifs that are present in proteins with completely different functions. Several genetic diseases in humans have been associated with mutations in their sequence, which has made them promising targets for rational drug design. The protein MJ0100 from Methanocaldococcus jannaschii includes a DUF39 domain of so far unknown function and a CBS domain pair (Bateman domain) at its C-terminus. This work presents the crystallographic analysis of four different states of the CBS motif pair of MJ0100 in complex with different numbers of S-adenosyl-L-methionine (SAM) and S methyl-5'-thioadenosine (MTA) ligands, providing evidence that ligand-induced conformational reorganization of Bateman domain dimers could be an important regulatory mechanism. These observations are in contrast to what is known from most of the other Bateman domain structures but are supported by recent studies on the magnesium transporter MgtE. Our structures represent the first example of a CBS domain protein complexed with SAM and/or MTA and might provide a structural basis for understanding the molecular mechanisms regulated by SAM upon binding to the C-terminal domain of human CBS, whose structure remains unknown. PMID- 20026079 TI - The recombinant amyloid-beta peptide Abeta1-42 aggregates faster and is more neurotoxic than synthetic Abeta1-42. AB - Aggregation of the amyloid-beta (Abeta) peptide is considered a central event in the pathogenesis of Alzheimer's disease (AD). In order to bypass methodological bias related to a variety of impurities commonly present in typical preparations of synthetic Abeta, we developed a simple, generally applicable method for recombinant production of human Abeta and Abeta variants in Escherichia coli that provides milligram quantities of Abeta in very high purity and yield. Amyloid fibril formation in vitro by human Abeta1-42, the key amyloidogenic Abeta species in AD, was completed threefold faster with recombinant Abeta1-42 compared to synthetic preparations. In addition, recombinant Abeta1-42 was significantly more toxic to cultured rat primary cortical neurons, and it was more toxic in vivo, as shown by strongly increased induction of abnormal phosphorylation of tau and tau aggregation into neurofibrillary tangles in brains of P301L tau transgenic mice. We conclude that even small amounts of impurities in synthetic Abeta-including a significant fraction of racemized peptides that cannot be avoided due to the technical limitations of peptide synthesis--prevent or slow Abeta incorporation into the regular quaternary structure of growing beta-amyloid fibrils. The results validate the use of recombinant Abeta1-42 for both in vitro and in vivo studies addressing the mechanisms underlying Abeta aggregation and its related biological consequences for the pathophysiology, therapy, and prevention of AD. PMID- 20026080 TI - Antitumor 1-nitroacridine derivative C-1748, induces apoptosis, necrosis or senescence in human colon carcinoma HCT8 and HT29 cells. AB - C-1748 is a DNA-binding agent with potent antitumor activity, especially towards prostate and colon carcinoma xenografts in mice. Here, we elucidated the nature of cellular response of human colon carcinoma HCT8 and HT29 cells to C-1748 treatment, at biologically relevant concentrations (EC(90) and their multiplicity). Cell cycle analysis showed gradual increase in HCT8 cells with sub G1 DNA content (25% after 72h) considered as apoptotic. Hypodiploid cell population increased up to 60% upon treatment with 4x EC(90) concentration of the drug. Compared with HCT8 cells, the fraction of sub-G1 HT29 cells did not exceed 14%, even following 4-fold dose escalation. Morphological changes and biochemical markers such as: phosphatydylserine externalization, apoptotic DNA breaks, mitochondrial dysfunction and caspase activation confirmed the presence of considerable amount of apoptotic HCT8 cells but only a low amount of apoptotic HT29 cells. Next, we demonstrated that HCT8 cells surviving after exposure to C 1748 were in the state of senescence, based on altered cell morphology and expression of a pH 6-dependent beta-galactosidase. On the contrary, no beta galactosidase staining was observed in HT29 cells after C-1748 treatment. Moreover, prolonged drug incubation (up to 168h) resulted in massive detachment of cells from culture plates, which together with Annexin V/PI results, indicated that necrosis was the main response of HT29 cells to C-1748 treatment. We also determined the ability of C-1748 to induce reactive oxygen species (ROS) in colon cancer cells and demonstrated, that generation of ROS was not essential for C 1748-induced apoptosis and cytotoxic activity of this drug. PMID- 20026081 TI - AMPK-mediated GSK3beta inhibition by isoliquiritigenin contributes to protecting mitochondria against iron-catalyzed oxidative stress. AB - Isoliquiritigenin (ILQ), a flavonoid compound originated from Glycyrrhiza species, is known to activate SIRT1. Arachidonic acid (AA) in combination with iron (a catalyst of auto-oxidation) leads cells to produce excess reactive species with a change in mitochondrial permeability transition. In view of the importance of oxidative stress in cell death and inflammation, this study investigated the potential of ILQ to protect cells against the mitochondrial impairment induced by AA+iron and the underlying basis for this cytoprotection. Treatment with ILQ inhibited apoptosis induced by AA+iron, as evidenced by alterations in the levels of the proteins associated with cell viability: ILQ prevented a decrease in Bcl-x(L), and cleavage of poly(ADP-ribose)polymerase and procaspase-3. Moreover, ILQ inhibited the ability of AA+iron to elicit mitochondrial dysfunction. In addition, superoxide generation in mitochondria was attenuated by ILQ treatment. Consistently, ILQ prevented cellular H2O2 production increased by AA+iron, thereby enabling cells to restore GSH content. ILQ treatment enhanced inhibitory phosphorylation of glycogen synthase kinase-3beta (GSK3beta), and prevented a decrease in the GSK3beta phosphorylation elicited by AA+iron, which contributed to protecting cells and mitochondria. GSK3beta phosphorylation by ILQ was preceded by AMP-activated protein kinase (AMPK) activation, which was also responsible for mitochondrial protection, as shown by reversal of its effect in the experiments using a dominant negative mutant of AMPK and compound C. Moreover, the AMPK activation led to GSK3beta phosphorylation. These results demonstrate that ILQ has the ability to protect cells from AA+iron-induced H2O2 production and mitochondrial dysfunction, which is mediated with GSK3beta phosphorylation downstream of AMPK. PMID- 20026082 TI - Rosiglitazone increases cell surface GLUT4 levels in 3T3-L1 adipocytes through an enhancement of endosomal recycling. AB - Insulin induces a translocation of the glucose transporter GLUT4 from intracellular storage compartments towards the cell surface in adipocytes and skeletal muscle cells, allowing the cells to take up glucose. In type 2 diabetes associated insulin resistance, the efficiency of this process is reduced. The thiazolidinediones, widely prescribed as anti-diabetic therapy, are generally regarded as insulin-sensitizers. The aim of this study was to evaluate the effect of the thiazolidinedione rosiglitazone (BRL 49653) on GLUT4 in adipocytes. When applied during differentiation, rosiglitazone dose dependently augmented GLUT4 expression along with the formation of lipid droplets. Intriguingly, its presence during differentiation led to increases in both cell surface GLUT4 levels and insulin sensitivity of GLUT4 translocation in mature adipocytes. Treatment of fully differentiated adipocytes with rosiglitazone also led to increases in GLUT4 at the plasma membrane. Rosiglitazone similarly affected cell surface levels of the endosomal transferrin receptor, but did not alter the GLUT4 internalization rate. The augmentation in cell surface GLUT4 levels was maintained in adipocytes that were rendered insulin-resistant in vitro by a 24h insulin treatment and moreover in these cells rosiglitazone also fully restored insulin-induced GLUT4 translocation. We conclude that in adipocytes, rosiglitazone increases cell surface GLUT4 levels by increasing its endosomal recycling and restores insulin induced GLUT4 translocation in insulin resistance. These results implicate novel modes of action on GLUT4 that are all likely to contribute to the insulin sensitizing effect of rosiglitazone in type 2 diabetes. PMID- 20026083 TI - Inhibition of the JAK-STAT3 pathway by andrographolide enhances chemosensitivity of cancer cells to doxorubicin. AB - Andrographolide (Andro), a diterpenoid lactone isolated from a traditional herbal medicine Andrographis paniculata, is known to possess potent anti-inflammatory and anticancer properties. In this study, we sought to examine the effect of Andro on signal transducer and activator of transcription 3 (STAT3) pathway and evaluate whether suppression of STAT3 activity by Andro could sensitize cancer cells to a chemotherapeutic drug doxorubicin. First, we demonstrated that Andro is able to significantly suppress both constitutively activated and IL-6-induced STAT3 phosphorylation and subsequent nuclear translocation in cancer cells. Such inhibition is found to be achieved through suppression of Janus-activated kinase (JAK)1/2 and interaction between STAT3 and gp130. For understanding the biological significance of the inhibitory effect of Andro on STAT3, we next investigated the effect of Andro on doxorubicin-induced apoptosis in human cancer cells. In our study the constitutive activation level of STAT3 was found to be correlated to the resistance of cancer cells to doxorubicin-induced apoptosis. Both the short-term MTT assay and the long-term colony formation assay showed that Andro dramatically promoted doxorubicin-induced cell death in cancer cells, indicating that Andro enhances the sensitivity of cancer cells to doxorubicin mainly via STAT3 suppression. These observations thus reveal a novel anticancer function of Andro and suggest a potential therapeutic strategy of using Andro in combination with chemotherapeutic agents for treatment of cancer. PMID- 20026084 TI - Emotion processing and theory of mind in schizophrenia patients and their unaffected first-degree relatives. AB - Previous studies have suggested that social cognition is affected in individuals with schizophrenia. The purpose of this study was to explore to what extent social cognition deficits are shared by unaffected first-degree relatives, and the nature of the relationship between performance in different paradigms of social cognition. 20 Schizophrenia patients (7 females, 31+/-10 years), 20 healthy age- and gender-matched individuals, 20 unaffected first-degree relatives of the schizophrenia patients (11 females, 50+/-20 years), and 20 healthy individuals matched for age and gender were recruited. Patients showed deficits in the detection of social Faux Pas (0.80+/-0.17 vs. controls: 0.94+/-0.09, p=0.025) and the correct identification of Theory of Mind stories (0.71+/-0.13 vs. controls: 0.82+/-0.12, p=0.038). Relatives performed poorly in the Faces Test (0.83+/-0.14 vs. controls: 0.9+/-0.08, p=0.048), the Reading the Mind in the Eyes Test (0.59+/-0.17 vs. controls: 0.71+/-0.14, p=0.046) and the detection of social Faux Pas (0.8+/-0.2 vs. controls: 0.93+/-0.09, p=0.024). Abnormalities were independent of age, years of education, and general cognitive performance in patients and their relatives. Performance in an Emotion Processing task (Faces Test) was correlated with performance in theory of mind tests in healthy individuals and relatives of patients with schizophrenia only. These results suggest that schizophrenia patients and their unaffected first-degree relatives display similar but nonidentical patterns of social cognition processing. PMID- 20026085 TI - Executive neuropsychological functioning in individuals with Williams syndrome. AB - The present study investigated executive neuropsychological functioning in individuals with the neuro-developmental disorder Williams syndrome (WS) using a set of validated standardized neuropsychological tasks. Relatively few studies have examined frontal lobe related executive functions within the cognitive phenotype associated with the disorder. The present study compared participants with WS to typically developing participants who were individually matched for (1) chronological age and (2) verbal mental age (N=19 each group) on tasks of attention-set shifting, planning and working memory from the Cambridge Neuropsychological Test Automated Battery (CANTAB). To address the specificity of executive function impairment, non-executive tasks of delayed short-term memory and short-term memory span were also administered. Individuals with WS (mean age 18 years) showed impaired executive functioning on tasks of attention set shifting, working memory, and planning. Non-executive deficits were also observed in short-term delayed memory and memory span. Neuropsychological impairments were correlated with a range of behavioural problems assessed using parent-rated Questionnaires. Overall, these findings point to the role of a range of executive function impairments in WS but further suggest that cognitive impairments extend beyond executive dysfunction. PMID- 20026086 TI - Errors recruit both cognitive and emotional monitoring systems: simultaneous intracranial recordings in the dorsal anterior cingulate gyrus and amygdala combined with fMRI. AB - We studied error monitoring in a human patient with unique implantation of depth electrodes in both the left dorsal cingulate gyrus and medial temporal lobe prior to surgery. The patient performed a speeded go/nogo task and made a substantial number of commission errors (false alarms). As predicted, intracranial Local Field Potentials (iLFPs) in dorsal anterior cingulate indexed the detection of errors, showing an early differential activity around motor execution for false alarms, relative to correct responses (either hits or correct inhibitions). More surprisingly, we found that the left amygdala also participated to error monitoring (although no emotional stimuli were used), but with a very different neurophysiological profile as compared with the dorsal cingulate cortex. Amygdala iLFPs showed a precise and reproducible temporal unfolding, characterized by an early monophasic response for correct hits around motor execution, which was delayed by approximately 300ms for errors (even though actual RTs were almost identical in these two conditions). Moreover, time-frequency analyses demonstrated a reliable and transient coupling in the theta band around motor execution between these two distant regions. Additional fMRI investigation in the same patient confirmed a differential involvement of the dorsal cingulate cortex vs. amygdala in error monitoring during this go/nogo task. Finally, these intracranial results for the left amygdala were replicated in a second patient with intracranial electrodes in the right amygdala. Altogether, these results suggest that the amygdala may register the motivational significance of motor actions on a trial-by-trial basis, while the dorsal anterior cingulate cortex may provide signals concerning failures of cognitive control and behavioral adjustment. More generally, these data shed new light on neural mechanisms underlying self-monitoring by showing that even "simple" motor actions recruit not only executive cognitive processes (in dorsal cingulate) but also affective processes (in amygdala). PMID- 20026087 TI - Action monitoring in children with or without a family history of ADHD--effects of gender on an endophenotype parameter. AB - Attention-deficit/hyperactivity disorder (ADHD) is a frequent and highly heritable disorder overrepresented in boys. In a recent study investigating boys only, we found that action monitoring deficits as reflected by certain behavioral and electrophysiological parameters were familially driven. As gender may also have an important impact, this was examined in the current study with nonaffected children aged 8-15 years having relatives suffering from ADHD (N=37, 21 female symbol) and with age-matched controls without family history of ADHD (N=33, 11 female symbol). Extending our previous findings that action monitoring is a potential endophenotype for boys with ADHD, familially driven deficits were confirmed independently of gender. Thus, despite sharing the phenotype with controls, nonaffected siblings showed ADHD-like impairments albeit of smaller magnitude. However, girls performed generally more accurately, which in turn may have produced the differences between nonaffected siblings and controls in affective error processing that were not present in our boys-only assessment. PMID- 20026090 TI - In vivo type 1 cannabinoid receptor mapping in the 6-hydroxydopamine lesion rat model of Parkinson's disease. AB - Type 1 cannabinoid (CB1) receptors are expressed in high concentrations in the central nervous system, including the basal ganglia, and could have direct or indirect effects on motor behavior through modulation of dopaminergic, glutamatergic and GABA-ergic neurotransmission. Using the CB1 receptor radioligand [(18)F]MK-9470 and small-animal PET, we investigated for the first time in vivo cerebral changes in [(18)F]MK-9470 binding in the 6-hydroxydopamine (6-OHDA) rat model of Parkinson's disease (PD), parallel to dopamine transporter (DAT) imaging, tyrosine hydroxylase (TH) staining, and behavioral measurements. In the 6-OHDA model, relative [(18)F]MK-9470 PET binding decreased in the contralateral cerebellum (-9%, p<0.0004) and caudate-putamen bilaterally (ipsilateral -8%, contralateral -7%; p=0.001 and p<0.0003, respectively). The number of TH(+) neurons in the substantia nigra was inversely correlated to CB1 receptor binding in the ipsilateral cerebellum (p=1.10(-6)). The behavioral outcome was positively related to regional CB1 receptor binding in the contralateral somatosensory cortex (p=4.10(-6)). In vivo [(18)F]MK-9470 PET imaging points to changes in endocannabinoid transmission, specifically for CB1 receptors in the 6-OHDA model of PD, with mainly involvement of the caudate putamen, but also distant regions of the motor circuitry, including the cerebellum and somatosensory cortex. PMID- 20026089 TI - Disparate developmental neurotoxicants converge on the cyclic AMP signaling cascade, revealed by transcriptional profiles in vitro and in vivo. AB - Cell-signaling cascades are convergent targets for developmental neurotoxicity of otherwise unrelated agents. We compared organophosphates (chlorpyrifos, diazinon), an organochlorine (dieldrin) and a metal (Ni(2+)) for their effects on neuronotypic PC12 cells, assessing gene transcription involved in the cyclic AMP pathway. Each agent was introduced during neurodifferentiation at a concentration of 30 microM for 24 or 72 h and we assessed 69 genes encoding adenylyl cyclase isoforms and regulators, G-protein alpha-and beta,gamma-subunits, protein kinase A subtypes and the phosphodiesterase family. We found strong concordance among the four agents across all the gene families, with the strongest relationships for the G-proteins, followed by adenylyl cyclase, and lesser concordance for protein kinase A and phosphodiesterase. Superimposed on this pattern, chlorpyrifos and diazinon were surprisingly the least alike, whereas there was strong concordance of dieldrin and Ni(2+) with each other and with each individual organophosphate. Further, the effects of chlorpyrifos differed substantially depending on whether cells were undifferentiated or differentiating. To resolve the disparities between chlorpyrifos and diazinon, we performed analyses in rat brain regions after in vivo neonatal exposures; unlike the in vitro results, there was strong concordance. Our results show that unrelated developmental neurotoxicants can nevertheless produce similar outcomes by targeting cell signaling pathways involved in neurodifferentiation during a critical developmental period of vulnerability. Nevertheless, a full evaluation of concordance between different toxicants requires evaluations of in vitro systems that detect direct effects, as well as in vivo systems that allow for more complex interactions that converge on the same pathway. PMID- 20026088 TI - Role of innate and drug-induced dysregulation of brain stress and arousal systems in addiction: Focus on corticotropin-releasing factor, nociceptin/orphanin FQ, and orexin/hypocretin. AB - Stress-like symptoms are an integral part of acute and protracted drug withdrawal, and several lines of evidence have shown that dysregulation of brain stress systems, including the extrahypothalamic corticotropin-releasing factor (CRF) system, following long-term drug use is of major importance in maintaining drug and alcohol addiction. Recently, two other neuropeptide systems have attracted interest, the nociceptin/orphanin FQ (N/OFQ) and orexin/hypocretin (Orx/Hcrt) systems. N/OFQ participates in a wide range of physiological responses, and the hypothalamic Orx/Hcrt system helps regulate several physiological processes, including feeding, energy metabolism, and arousal. Moreover, these two systems have been suggested to participate in psychiatric disorders, including anxiety and drug addiction. Dysregulation of these systems by chronic drug exposure has been hypothesized to play a role in the maintenance of addiction and dependence. Recent evidence demonstrated that interactions between CRF-N/OFQ and CRF-Orx/Hcrt systems may be functionally relevant for the control of stress-related addictive behavior. The present review discusses recent findings that support the hypotheses of the participation and dysregulation of these systems in drug addiction and evaluates the current understanding of interactions among these stress-regulatory peptides. PMID- 20026091 TI - Localization of CXCR4 in the forebrain of the adult rat. AB - Chemokines are small secreted proteins that act as chemoattractants, and their role as neuromodulators in the brain has recently been appreciated. CXCL12 is one of the few chemokines found in neurons and expressed constitutively in the central nervous system. Previous data from our laboratory demonstrate the ability of CXCL12 to modulate the behavioral effects of cocaine, and this modulation is dependent on the central site of administration of CXCL12. The present study used single-staining immunohistochemical and dual-staining immunofluorescent methods to determine the localization of the CXCL12 receptor, CXCR4, in the caudate putamen and nucleus accumbens of the adult rat brain. Results demonstrated that individual neurons in both the caudate putamen and lateral shell of the nucleus accumbens express both CXCR4 and D1 dopamine receptors. Immunofluorescent studies showed that CXCR4 was co-expressed with ChAT, a marker for cholinergic neurons, and with GAD C38, a marker for GABAergic neurons, in the caudate putamen and lateral shell of the accumbens. No evidence of CXCR4 was found in the medial shell or core regions of the nucleus accumbens. These data demonstrate that CXCR4 is expressed by subpopulations of cholinergic and GABAergic neurons in the striatum and suggest that CXCR4 is well-positioned to modulate striatal function. PMID- 20026092 TI - Testosterone exposure, dopaminergic reward, and sensation-seeking in young men. AB - To test the relationship between androgen exposure, dopaminergic reward and sensation-seeking, we compared variation in salivary testosterone (T), 2D:4D digit ratio, facial masculinity, Zuckerman's sensation-seeking scale (SSS) and the D4 dopamine receptor (DRD4) genes from 98 young men, between the ages of 18 and 23 years. In univariate analyses, both salivary T and facial masculinity were significantly correlated with the SSS boredom susceptibility subscale, while the presence of the 7-repeat allele (7R+) in the dopamine receptor D4 gene was associated with the SSS thrill and adventure-seeking and overall sensation seeking. Neither left nor right 2D:4D digit ratio was associated with any sensation-seeking scale. In multivariate models, salivary T and facial masculinity were significant predictors of SSS boredom susceptibility, while 7R+ was a significant predictor of SSS thrill and adventure-seeking. For overall SSS, both 7R+ and salivary T were significant predictors. There was no significant interaction of 7R+ and androgen exposure for SSS or any of the SSS subscales. These results add to earlier reports of an association between T and sensation seeking. In addition, our results also indicate that genetic variation in DRD4 is independently associated with SSS sensation-seeking. PMID- 20026093 TI - Analysis of hydroquinone and catechol in peripheral blood of benzene-exposed workers. AB - We have developed a gas chromatography-mass spectrometry method for analysis of benzene (BZ) metabolites in human urine and blood. Here we describe peripheral blood concentrations of hydroquinone (HQ(1)) and catechol (CAT(2)) in total, protein-bound, and unbound (free) forms obtained from BZ-exposed factory workers and controls. Total and unbound metabolites were directly measured in independent experiments, while bound forms were calculated as [total]-[unbound]. In this subset of a larger study, breathing zone benzene, toluene, and xylene were measured for the duration of a workshift, and end-shift blood samples taken from 143 subjects and controls. Potential lifestyle and environmental influences were assessed by questionnaire and bioassay, and single nucleotide polymorphisms in xenobiotic metabolizing enzymes NQO1, MPO, CYP2E1, and GSTT1 were also analyzed for potential contribution to differences in blood metabolite concentration. Total CAT, bound CAT, total HQ, and bound HQ correlated well with benzene exposure, while unbound CAT and HQ displayed no correlation. Nearly all of the metabolites found in blood were bound to protein (CAT 96-99+%, HQ 78-92+%), and when the ratio of bound to unbound metabolites were compared in subsets of exposed workers, the increase in blood metabolite concentration was nearly all due to an increase in the protein-bound molecule. These findings suggest that a threshold for conjugation does not exist within the exposure spectrum studied (0.01-78.8 mg/m(3)). This method demonstrates the feasibility of analyzing benzene metabolites in human blood, and should allow for further investigation of the health effects of benzene and its metabolites. PMID- 20026094 TI - Systems biology of human benzene exposure. AB - Toxicogenomic studies, including genome-wide analyses of susceptibility genes (genomics), gene expression (transcriptomics), protein expression (proteomics), and epigenetic modifications (epigenomics), of human populations exposed to benzene are crucial to understanding gene-environment interactions, providing the ability to develop biomarkers of exposure, early effect and susceptibility. Comprehensive analysis of these toxicogenomic and epigenomic profiles by bioinformatics in the context of phenotypic endpoints, comprises systems biology, which has the potential to comprehensively define the mechanisms by which benzene causes leukemia. We have applied this approach to a molecular epidemiology study of workers exposed to benzene. Hematotoxicity, a significant decrease in almost all blood cell counts, was identified as a phenotypic effect of benzene that occurred even below 1 ppm benzene exposure. We found a significant decrease in the formation of progenitor colonies arising from bone marrow stem cells with increasing benzene exposure, showing that progenitor cells are more sensitive to the effects of benzene than mature blood cells, likely leading to the observed hematotoxicity. Analysis of transcriptomics by microarray in the peripheral blood mononuclear cells of exposed workers, identified genes and pathways (apoptosis, immune response, and inflammatory response) altered at high (>10 ppm) and low (<1 ppm) benzene levels. Serum proteomics by SELDI-TOF-MS revealed proteins consistently down-regulated in exposed workers. Preliminary epigenomics data showed effects of benzene on the DNA methylation of specific genes. Genomic screens for candidate genes involved in susceptibility to benzene toxicity are being undertaken in yeast, with subsequent confirmation by RNAi in human cells, to expand upon the findings from candidate gene analyses. Data on these and future biomarkers will be used to populate a large toxicogenomics database, to which we will apply bioinformatic approaches to understand the interactions among benzene toxicity, susceptibility genes, mRNA, and DNA methylation through a systems biology approach. PMID- 20026095 TI - Chronic ingestion of 2-deoxy-D-glucose induces cardiac vacuolization and increases mortality in rats. AB - Calorie restriction (CR), the purposeful reduction of energy intake with maintenance of adequate micronutrient intake, is well known to extend the lifespan of laboratory animals. Compounds like 2-deoxy-D-glucose (2DG) that can recapitulate the metabolic effects of CR are of great interest for their potential to extend lifespan. 2DG treatment has been shown to have potential therapeutic benefits for treating cancer and seizures. 2DG has also recapitulated some hallmarks of the CR phenotype including reduced body temperature and circulating insulin in short-term rodent trials, but one chronic feeding study in rats found toxic effects. The present studies were performed to further explore the long-term effects of 2DG in vivo. First we demonstrate that 2DG increases mortality of male Fischer-344 rats. Increased incidence of pheochromocytoma in the adrenal medulla was also noted in the 2DG treated rats. We reconfirm the cardiotoxicity of 2DG in a 6-week follow-up study evaluating male Brown Norway rats and a natural form of 2DG in addition to again examining effects in Fischer 344 rats and the original synthetic 2DG. High levels of both 2DG sources reduced weight gain secondary to reduced food intake in both strains. Histopathological analysis of the hearts revealed increasing vacuolization of cardiac myocytes with dose, and tissue staining revealed the vacuoles were free of both glycogen and lipid. We did, however, observe higher expression of both cathepsin D and LC3 in the hearts of 2DG-treated rats which indicates an increase in autophagic flux. Although a remarkable CR-like phenotype can be reproduced with 2DG treatment, the ultimate toxicity of 2DG seriously challenges 2DG as a potential CR mimetic in mammals and also raises concerns about other therapeutic applications of the compound. PMID- 20026096 TI - Use of human bronchial epithelial cells (BEAS-2B) to study immunological markers resulting from exposure to PM(2.5) organic extract from Puerto Rico. AB - Fine particulate air pollutants, mainly their organic fraction, have been demonstrated to be associated with cardiovascular and respiratory health problems. Puerto Rico has been reported to have the highest prevalence of pulmonary diseases (e.g., asthma) in the United States. The aim of this study was to assess, for the first time, the immunological response of human bronchial epithelial cells (BEAS-2B) to organic extracts isolated from airborne particulate matter (PM(2.5)) in Puerto Rico. Organic extracts from PM(2.5) collected throughout an 8-month period (2000-2001) were pooled (composite) in order to perform chemical analysis and biological activity testing. BEAS-2B cells were exposed to PM(2.5) organic extract to assess cytotoxicity, levels of cytokines and relative gene expression of MHC-II, hPXR and CYP3A5. Our findings show that organic PM(2.5) consist of toxic as well as bioactive components that can regulate the secretion of cytokines in BEAS-2B, which could modulate inflammatory response in the lung. Trace element analyses confirmed the presence of metals in organic extracts highlighting the relative high abundance of Cu and Zn in polar organic extracts. Polar organic extracts exhibited dose-dependant toxicity and were found to significantly induce the release of interleukin 6 (IL-6), IL-1beta and IL-7 while significantly inhibiting the secretion of IL-8, G-CSF and MCP-1. Moreover, MHC-II transcriptional activity was up-regulated after 24 h of exposure, whereas PXR and CYP3A5 were down-regulated. This research provides a new insight into the effects of PM(2.5) organic fractions on specific effectors and their possible role in the development of respiratory inflammatory diseases in Puerto Rico. PMID- 20026098 TI - HER2 codon 655 polymorphism is associated with advanced uterine cervical carcinoma. AB - OBJECTIVES: It has been suggested that overexpression of HER2 in advanced cervical tumors can be considered an independent predictor of poor patient outcome. DESIGN AND METHODS: Employing PCR-RFLPs, we examined the distribution of HER2 Ile655Val (rs 1136201) genotypes and alleles in patients with advanced cervical cancer (n=109) and controls (n=220). RESULTS: Odds ratio (OR) for patients with advanced cervical cancer with the HER2 Val/Val homozygous or Val/Ile heterozygous state was 1.778 (95% CI=1.117-2.830, p=0.0176). We also observed an association of the HER2 Val/Val genotype with advanced cervical cancer in the patient group OR=3.706 (95% CI=1.061-12.950, p=0.0459). However, we did not find a significant association between the distribution of genotypes or alleles and cancer characteristics for the HER2 Ile655Val polymorphism. CONCLUSIONS: Our results indicate that the HER2 655Val variant may be associated with the incidence of advanced cervical cancer. PMID- 20026097 TI - Identification of myocardial injury in the emergency setting. AB - Within the past decade, the use of biomarkers to detect myocardial injury in the emergency department (ED) has been given increasing prominence as evident by the numerous studies and guidelines documenting their use. This review details the scope of the clinical problem, the history of changes in the definition of myocardial infarction (MI) and the new approaches, as well as suggestions for using laboratory biomarkers in the early detection of MI in the ED. PMID- 20026099 TI - The relationship between paraoxonase1-192 polymorphism and activity with coronary artery disease. AB - OBJECTIVE: We tested the association between PON1 polymorphism, PON1 activity, oxidative susceptibility of LDL and coronary artery disease in Egyptians. METHODS: PON1 polymorphism, serum PON1 activity, lipoprotein oxidation susceptibility and lipid profile were measured. RESULTS: Levels of HDL and paraoxonase activity were significantly decreased in CAD patients compared to control group, and in patients with three vessels compared to those of single or two vessels disease. High-activity allele (R) has a more atherogenic lipid profile than for the low activity allele (Q). PON1 RR genotype has nine fold risks to develop CAD in Egyptians while those with PON1 QR genotype have four fold risks. CONCLUSION: The PON1 activity is lower in subject with CAD and there is a significant relationship between activity of PON1 and the severity of coronary atherosclerosis. Also, we provide evidence of a significant association between R allele of the PON1 polymorphism and the development of coronary artery disease. PMID- 20026100 TI - Effect of second and third generation oral contraceptives on C-reactive protein, lipids and apolipoproteins in young, non-obese, non-smoking apparently healthy women. AB - OBJECTIVE: Third generation oral contraceptives (OC) may increase the risk of cardiovascular events in healthy young women. DESIGN AND METHODS: We assessed the effect of second and third generation OC on CRP, lipids and apolipoproteins in 128 women. RESULTS: CRP was significantly higher in third generation contraceptive users. The main determinant of CRP in OC users was triglycerides. CONCLUSIONS: Young women using oral contraceptives, especially third generation formulas, might not be free of cardiovascular risk having increased CRP concentration. PMID- 20026101 TI - Azaspiracid poisoning (AZP) toxins in shellfish: toxicological and health considerations. AB - It has been almost a decade since a previously unknown human toxic syndrome, azaspiracid poisoning (AZP), emerged as the cause of severe gastrointestinal illness in humans after the consumption of mussels (Mytilus edulis). Structural studies indicated that these toxins, azaspiracids, were of a new unprecedented class containing novel structural features. It is now known that the prevalent azaspiracids in mussels are AZA1, AZA2 and AZA3, which differ from each other in their degree of methylation. Several hydroxylated and carboxylated analogues of the main azaspiracids have also been identified, presumed to be metabolites of the main toxins. Since its first discovery in Irish mussels, the development of facile sensitive and selective LC-MS/MS methods has resulted in the discovery of AZA in other countries and in other species. Mice studies indicate that this toxin class can cause serious tissue injury, especially to the small intestine, and chronic exposure may increase the likelihood of the development of lung tumours. Studies also show that tissue recovery is very slow following exposure. These observations suggest that AZA is more dangerous than the other known classes of shellfish toxins. Consequently, in order to protect human consumers, proper risk assessment and regulatory control of shellfish and other affected species is of the utmost importance. PMID- 20026102 TI - Vitrification of porcine articular cartilage. AB - The limited availability of fresh osteochondral allograft tissues necessitates the use of banking for long-term storage. A vitrification solution containing a 55% cryoprotectant formulation, VS55, previously studied using rabbit articular cartilage, was evaluated using porcine articular cartilage. Specimens ranging from 2 to 6 mm in thickness were obtained from 6mm distal femoral cartilage cores and cryopreserved by vitrification or freezing. The results of post-rewarming viability assessments employing alamarBlue demonstrated a large decrease (p<0.001) in viability in all three sizes of cartilage specimen vitrified with VS55. This is in marked contrast with prior experience with full thickness, 0.6 mm rabbit cartilage. Microscopic examination following cryosubstitution confirmed ice formation in the chondrocytes of porcine cartilage vitrified using VS55. Experiments using a more concentrated vitrification formulation (83%), VS83, showed a significant treatment benefit for larger segments of articular cartilage. Differences between the VS55 and the VS83 treatment groups were significant at p<0.001 for 2 mm and 4 mm plugs, and at p<0.01 for full thickness, 6 mm plugs. The percentage viability in fresh controls, compared to VS55 and VS83, was 24.7% and 80.7% in the 2 mm size group, 18.2% and 55.5% in the 4 mm size group, and 5.2% and 43.6% in the 6 mm group, respectively. The results of this study continue to indicate that vitrification is superior to conventional cryopreservation with low concentrations of dimethyl sulfoxide by freezing for cartilage. The vitrification technology presented here may, with further process development, enable the long-term storage and transportation of living cartilage for repair of human articular surfaces. PMID- 20026104 TI - Xylans from Scinaia hatei: Structural features, sulfation and anti-HSV activity. AB - Natural compounds offer interesting pharmacological perspectives for antiviral drug development with regard to broad-spectrum antiviral properties and novel modes of action. In this study, we have analyzed alkali-extracted xylan of Scinaia hatei. Alkali extraction of this red alga yielded a xylan shown to have a molecular mass of 120kDa and a linear structure of (1-->4)-linked beta-d xylopyranosyl residues. Derivatives (S1, S2, S3 and S4) generated by chemical sulfation from this macromolecule had degree of sulfation between 0.93 and 1.95, and contained strong anti-HSV activity with inhibitory concentration 50% (IC(50)) from 0.22 to 1.37mug/ml. Furthermore, they had no direct inactivating effect on virions in a virucidal assay. Sulfate groups account for their in vitro antiviral activity. Interestingly, sulfated xylans already exerted anti-HSV activity when only pre-incubated with the cultured cells prior to infection, thus pointing to a main inhibitory effect on viral entry. PMID- 20026103 TI - The effects of maternal smoking during pregnancy on offspring outcomes. AB - OBJECTIVE: To evaluate the possible association between maternal smoking during pregnancy and offspring outcomes of birth weight, pre-term birth, remediation, low scholastic achievement, regular smoking, attention deficit hyperactivity disorder and conduct problems while controlling for similar behaviors in parents. METHODS: Using telephone interviews, data were collected, in 2001 and 2004, as a part of two United States offspring-of-twins projects. Fathers, who were twins participating in the Vietnam Era Twin Registry, their female spouse and their offspring were interviewed - information on 1,342 unique pregnancies in mothers with a history of regular smoking was utilized for these analyses. The association between maternal smoking during pregnancy and birth weight, pre-term birth, remediation, low scholastic achievement, regular smoking, attention deficit hyperactivity disorder and conduct disorder while controlling for similar behaviors in parents, was examined using regression. RESULTS: Maternal smoking during pregnancy was associated with decreased birth weight, low scholastic achievement, regular smoking and attention deficit hyperactivity disorder. However, the association between maternal smoking during pregnancy and offspring attention deficit hyperactivity disorder was explained by maternal attention deficit hyperactivity disorder. Maternal smoking during pregnancy was also associated with earlier age of offspring initiation of smoking and onset of regular smoking. CONCLUSIONS: Maternal smoking during pregnancy may influence certain offspring outcomes via mechanisms that are independent from genetic risk attributable to comorbid conditions. Assisting expecting mothers with their smoking cessation efforts will likely provide widespread health benefits to both mother and offspring. PMID- 20026105 TI - Sequence and structural analysis of binding site residues in protein-protein complexes. AB - The binding sites in protein-protein complexes have been identified with different methods including atomic contacts, reduction in solvent accessibility and interaction energy between the interacting partners. In our earlier work, we have developed an energy-based criteria for identifying the binding sites in protein-protein complexes, which showed that the interacting residues are different from that obtained with distance-based methods. In this work, we analyzed the binding site residues based on sequence and structural properties, such as, neighboring residues, secondary structure, solvent accessibility, conservation of residues, medium and long-range contacts and surrounding hydrophobicity. Our results showed that the neighboring residues of binding sites in proteins and ligands are different from each other although the interacting pairs of residues have a common behavior. The analysis on surrounding hydrophobicity reveals that the binding residues are less hydrophobic than non binding sites, which suggests that the hydrophobic core are important for folding and stability whereas the surface seeking residues play a critical role in binding. This tendency has been verified with the number of contacts in binding sites. In addition, the binding site residues are highly conserved compared with non-binding residues. We suggest that the incorporation of sequence and structure based features may improve the prediction accuracy of binding sites in protein protein complexes. PMID- 20026106 TI - Prevalence, nucleotide sequence and expression studies of two proteins of a 5.6kb, class III, Bacteroides plasmid frequently found in clinical isolates from European countries. AB - In order to gain information on the significance and functions of small molecular weight plasmids (usually regarded as cryptic) of Bacteroides isolates, we screened 178 European clinical Bacteroides isolates for plasmids and determined the nucleotide sequence of a 5.6kb plasmid. The previously observed frequent plasmid types (Classes I-III; 2.7, 4.2 and 5.6kb, respectively) were found to exhibit different distributions in five European countries as concerns plasmid type, geographical location, taxonomy and time course. The Class I plasmids displayed markedly different distribution properties from those of the Class II and III plasmids. The nucleotide sequence of a representative of the most frequent Class III plasmids, pBFP35, originating from Hungary, was determined (5594bp) and analyzed. A total of eight open reading frames (ORFs) were annotated, of which four proved to participate unequivocally in such plasmid maintenance functions as replication (repA(P35)), mobilization (mobA(P35)) and stability (mazE(P35) and mazF(P35)). Four additional ORFs (orf1-4) were identified. Orf1 was predicted to code a lipoprotein. In expression studies in an Escherichia coli host, Orf1 behaved as a periplasmic protein. PMID- 20026107 TI - Death receptors: targets for cancer therapy. AB - Apoptosis is the cell's intrinsic program to death, which plays an important role in physiologic growth control and homeostasis. Apoptosis can be triggered by death receptors (DRs), without any adverse effects. DRs are the members of tumor necrosis factor (TNF) receptor superfamily, known to be involved in apoptosis signaling, independent of p53 tumor-supressor gene. Selective triggering of DR mediated apoptosis in cancer cells is a novel approach in cancer therapy. So far, the best characterized DRs are CD95 (Fas/Apo1), TNF-related apoptosis-inducing ligand receptor (TRAILR) and tumor necrosis factor receptor (TNFR). Among these, TRAILR is emerging as most promising agent for cancer therapy, because it induces apoptosis in a variety of tumor and transformed cells without any toxicity to normal cells. TRAIL treatment in combination with chemotherapy or radiotherapy enhances TRAIL sensitivity or reverses TRAIL resistance by regulating downstream effectors. This review covers the current knowledge about the DRs, summarizes main signaling in DRs and also summarizes the preclinical approaches of these DRs in cancer therapy. PMID- 20026108 TI - Impairment of VEGF-A-stimulated lamellipodial extensions and motility of vascular endothelial cells by chondromodulin-I, a cartilage-derived angiogenesis inhibitor. AB - Chondromodulin-I (ChM-I) is a cartilage-derived angiogenesis inhibitor that has been identified as inhibitory to the growth activity of vascular endothelial cells. In our present study, we demonstrate the anti-angiogenic activity of recombinant human ChM-I (rhChM-I) in mouse corneal angiogenesis and examine its action. We focus on the VEGF-A-induced migration of vascular endothelial cells, a critical regulatory step in angiogenesis. In a modified Boyden chamber assay, nanomolar concentrations of rhChM-I inhibited the chemotactic migration of human umbilical vein endothelial cells (HUVECs) induced by VEGF-A as well as by FGF-2 and IGF-I. The ChM-I action was found to be endothelial cell-specific and independent of cell adhesions. Time-lapse analysis further revealed that rhChM-I markedly reduces VEGF-A-stimulated motility of HUVECs and causes frequent alterations of the moving front due to the appearance of multiple transient protrusions. This action involved the inhibition of cell spreading and the disrupted reorganization of the actin cytoskeleton upon VEGF-A stimulation. Consistent with these observations, rhChM-I was found to significantly reduce the activity of Rac1/Cdc42 during cell spreading, and the VEGF-A-induced Rac1 activity but not its basal activity in quiescent cells. Taken together, our present data suggest that ChM-I impairs the VEGF-A-stimulated motility of endothelial cells by destabilizing lamellipodial extensions. PMID- 20026109 TI - Organization of the lamin scaffold in the internal nuclear matrix of normal and transformed hepatocytes. AB - Nuclear lamins are among the more abundant proteins making up the internal nuclear matrix, but very little is known about their structure in the nucleoplasm. Using immunoelectron microscopy, we demonstrate the organization of lamins in the nuclear matrix isolated from rat hepatocytes for the first time. Lamin epitopes are arrayed both in locally ordered clusters and in quasi-regular rows. Fourier filtering of the images demonstrates that the epitopes are placed at the nodes and halfway between the nodes of square or rhombic lattices that are about 50 nm on each side, as well as along rows at regular approximately 25-nm intervals. In addition, we have compared this structure with that of the internal nuclear matrix isolated from persistent hepatocyte nodules. In transformed hepatocytes, the islands of lamin lattice are lost, and only a partial regularity in the rows of gold particles remains. We suggest that orthogonal lattice assembly might be an intrinsic property of lamin molecules, and that the disassembly may be triggered by simple molecular events such as phosphorylation. PMID- 20026111 TI - Septo-hippocampal networks in chronic epilepsy. AB - The medial septum inhibits the appearance of interictal spikes and seizures through theta rhythm generation. We have determined that medial septal neurons increase their firing rates during chronic epilepsy and that the GABAergic neurons from both medial and lateral septal regions are highly and selectively vulnerable to the epilepsy process. Since the lateral septal region receives a strong projection from the hippocampus and its neurons are vulnerable to epilepsy, their functional properties are probably altered by this disorder. Using the pilocarpine model of temporal lobe epilepsy we examined the pilocarpine induced functional alterations of lateral septal neurons and provided additional observations on the pilocarpine-induced functional alterations of medial septal neurons. Simultaneous extracellular recordings of septal neurons and hippocampal field potentials were obtained from chronic epileptic rats under urethane anesthesia. Our results show that: (1) the firing rates of lateral septal neurons were chronically decreased by epilepsy, (2) a subset of lateral septal neurons increased their firing rates before and during hippocampal interictal spikes, (3) the discharges of those lateral septal neurons were well correlated to the hippocampal interictal spikes, (4) in contrast, the discharges of medial septal neurons were not correlated with the hippocampal interictal spikes. We conclude that epilepsy creates dysfunctional and uncoupled septo-hippocampal networks. The elucidation of the roles of altered septo-hippocampal neuronal populations and networks during temporal lobe epilepsy will help design new and effective interventions dedicated to reduce or suppress epileptic activity. PMID- 20026112 TI - Effects of G-CSF treatment on neutrophil mobilization and neurological outcome after transient focal ischemia. AB - Several recent studies demonstrated beneficial effects of G-CSF treatment (granulocyte colony-stimulating factor) in various CNS disease. Possible mechanisms underlying this activity are neuroprotection, anti-apoptosis, angiogenesis and anti-inflammation. Hence, we investigated the efficacy of G-CSF administration in experimental stroke by determining infarct volume and neurological score in wildtype, G-CSF-deficient and G-CSF-treated G-CSF-deficient mice. Besides, cerebral ischemia is followed by an upregulation of endothelial adhesion molecules which promote leukocyte recruitment to the injured area. In combination with G-CSF-induced leukocytosis, increased peripheral neutrophils could aggregate within microvasculature and additionally impair blood perfusion of the ischemic tissue. Therefore, we analyzed the neutrophil counts in both vessel and tissue compartment 2 and 5 days post-stroke by immunohistochemistry. Here we show that G-CSF deficiency leads to increased infarct volumes, whereas G CSF substitution revokes detrimental effects by reducing lesion size and enhancing neurological outcome compared to untreated animals. Administration of G CSF is accompanied by significant increase of circulating neutrophils 2 days post ischemia but leukocytosis is restricted to the vessel compartment and has no deleterious effect on lesion formation and functional recovery. These observations are likely to be important for therapeutic targeting of G-CSF mediated neuroprotection in stroke. PMID- 20026110 TI - Neurobehavioral evidence for changes in dopamine system activity during adolescence. AB - Human adolescence has been characterized by increases in risk-taking, emotional lability, and deficient patterns of behavioral regulation. These behaviors have often been attributed to changes in brain structure that occur during this developmental period, notably alterations in gray and white matter that impact synaptic architecture in frontal, limbic, and striatal regions. In this review, we provide a rationale for considering that these behaviors may be due to changes in dopamine system activity, particularly overactivity, during adolescence relative to either childhood or adulthood. This rationale relies on animal data due to limitations in assessing neurochemical activity more directly in juveniles. Accordingly, we also present a strategy that incorporates molecular genetic techniques to infer the status of the underlying tone of the dopamine system across developmental groups. Implications for the understanding of adolescent behavioral development are discussed. PMID- 20026113 TI - Development of animal model for vasculatic neuropathy: Induction by ischemic reperfusion in the rat femoral artery. AB - Ischemic-reperfusion (I/R) is common in various pathological conditions like diabetic complication, complex regional pain syndrome type II (CRPS II), necrotizing vascular occlusive disease and trauma. We have developed an animal model of ischemic-reperfusion injury induced nociceptive sensory neuropathy in rats. The model was validated after 2, 4 and 6h of ischemia followed by prolonged reperfusion. The sensory behavioral assessment revealed thermal and mechanical hyperalgesia in paw and in tail which expressed the peripheral and central neuropathic pain respectively. We observed a decrease in the serum IL-10 and nerve conduction velocity and increase in the serum nitrate, malondialdehyde (MDA) and TNF-alpha levels in the 4 and 6h I/R groups in biochemical and electrophysiological evaluations. Histopathological study had revealed the decrease in nerve fiber density in the moderate and severe I/R groups. We selected the moderate (4h) ischemic-reperfusion injury as beneficial model because of the good correlation with clinical status for the development of neuropathy in human associated with severe pain disorders. This model can be used to explore pathophysiological mechanisms implied in the genesis of neuropathic pain and also to evaluate the new analgesic agents, peripheral neuro-vasoactive substances and neuroprotective drugs. PMID- 20026114 TI - Effects of chronic metabolic acidosis on splanchnic protein turnover and oxygen consumption in human beings. AB - BACKGROUND & AIMS: Although metabolic acidosis stimulates protein catabolism, its effects on splanchnic protein turnover and energy expenditure have not been measured in human beings. We investigated the effects of chronic metabolic acidosis (CMA) on splanchnic protein dynamics and oxygen consumption in human beings by using a leucine tracer and mass-balance techniques. METHODS: Five subjects were studied after 6 days of HCl-, CaCl(2)-, and NH(4)Cl-induced acidosis; 8 subjects served as controls. Blood samples were collected from the radial artery and the hepatic veins. Measurements were performed on plasma and whole-blood samples. RESULTS: Based on plasma measurements, subjects who had undergone CMA had lower rates of splanchnic proteolysis (-35%) and protein synthesis (-50%; P < .05) than controls, as well as a negative leucine kinetic balance (-6.81 +/- 2.48 micromol/kg/min/1.73 m(2) body surface [BS](-1)), compared with the neutral balance in control plasma samples (0.76 +/- 2.11 micromol/kg/min/1.73; P < .05 between groups). Based on measurements from whole blood, splanchnic proteolysis and protein synthesis did not differ significantly between CMA and control samples, and the net leucine kinetic balance was neutral in both groups (CMA, -0.69 +/- 1.57; controls, -0.74 +/- 3.45 micromol/kg/min/1.73). In CMA whole-blood measurements, splanchnic oxygen consumption (44.8 +/- 4.3 mL/min/1.73 m(2) BS) was slightly lower than in controls (57.5 +/- 8.4 mL/min/1.73 m(2) BS; P = NS). Splanchnic protein synthesis correlated with oxygen consumption (r = 0.82; P < .001). CONCLUSIONS: CMA reduces splanchnic protein turnover and results in a negative leucine balance--an effect that apparently is offset by the contribution of blood cells to organ leucine (and protein) dynamics. Protein synthesis is a major contributor (about 67%) to energy expenditure in splanchnic organs. PMID- 20026115 TI - Epithelial to mesenchymal transition is impaired in colon cancer cells with microsatellite instability. AB - BACKGROUND & AIMS: Colorectal cancers (CRCs) displaying DNA microsatellite instability (MSI) are associated with a favorable natural history, but the molecular basis for this observation has not been defined. We sought to determine whether the epithelial to mesenchymal transition (EMT) is impaired in MSI positive CRCs that characteristically have a mutant transforming growth factor beta receptor type II (TGFBR2) gene. METHODS: The induction of EMT by transforming growth factor-beta1 (TGF-beta1) was analyzed by phase contrast microscopy, immunofluorescence, quantitative reverse transcription polymerase chain reaction, immunoblotting, and cellular migration, and invasion assays. Expression of EMT markers was evaluated by immunohistochemistry and quantitative reverse transcription polymerase chain reaction in a series of human colorectal tumors. RESULTS: TGF-beta1 induced changes in cellular morphology, gene expression, motility, and invasion consistent with EMT in microsatellite stable (MSS) colon cancer cells, whereas cells with MSI and mutant TGFBR2 were unresponsive to TGF-beta1. These effects did not require Smad4, but depended on the recruitment of extracellular signal-regulated kinase. Tumor cells with MSI but wild-type TGFBR2 underwent EMT in response to TGF-beta1, indicating that TGFBR2 genotype is a key determinant of the EMT response in tumors with MSI. In human colorectal tumors, expression of EMT markers was significantly associated with adverse clinicopathologic features and the absence of MSI. CONCLUSIONS: These findings define a unique genotype-phenotype relationship between TGFBR2 and EMT that may contribute to the improved prognosis consistently observed in colon cancers with MSI. PMID- 20026116 TI - Neutrophil migration during liver injury is under nucleotide-binding oligomerization domain 1 control. AB - BACKGROUND & AIMS: A more complete understanding of the mechanisms involved in pathogen-associated molecular pattern signaling is crucial in the setting of liver injury. In intestinal diseases, nucleotide-binding oligomerization domain 1 (NOD1), a receptor for bacteria, appears to regulate cross-talk between innate and adaptive immunity, involving polymorphonuclear neutrophils (PMNs). Our aim was to explore the role of NOD1 in PMN-induced liver injury. METHODS: Nod1(+/+) and Nod1(-/-) mice were challenged with carbon tetrachloride (CCl(4)). Migration and phagocytosis of Nod1(+/+) and Nod1(-/-) PMN were studied in vivo and ex vivo. We evaluated main inflammatory pathways in PMNs by Western blot and CD11b expression using fluorescence-activated cell sorting. Mice were submitted to liver ischemia/reperfusion. RESULTS: After CCl(4) exposure, livers of Nod1(-/-) mice had more than 50% less PMN infiltration within necrotic areas than those of Nod1(+/+). PMNs isolated from Nod1(-/-) mice displayed a 90% decrease in migration capacity compared with Nod1(+/+) PMNs, whereas FK 565, a potent NOD1 ligand, increased PMN migration. Upon FK 565 stimulation, mitogen-activated protein kinase and nuclear factor kappaB were activated in Nod1(+/+) PMNs, but less so in Nod1(-/-) PMNs. Expression of CD11b on the Nod1(-/-) PMN was decreased compared with Nod1(+/+). The phagocytic capacity of Nod1(-/-) PMNs was decreased by more than 50% compared with Nod1(+/+). In an ischemia/reperfusion model of PMN induced liver injury, FK 565 increased lesions, whereas Nod1(-/-) mice were protected. CONCLUSIONS: The identification of NOD1 as a modulator of PMN function and migration in the liver suggests that this receptor may represent a new therapeutic target in PMN-dependent liver diseases. PMID- 20026117 TI - Ion channels in T lymphocytes: an update on facts, mechanisms and therapeutic targeting in autoimmune diseases. AB - During the last quarter of a century a large body of evidence was gathered about the involvement of ion channels in T lymphocyte activation. A series of remarkable findings promoted T cell ion channels to become potential pharmaceutical targets in the therapy of autoimmune disorders. Numerous comprehensive reviews describe the types of ion channels found in the plasma membrane of T cells and their roles in signaling pathways leading to activation, the changes in the expression of these channels brought upon by differentiation to various T cell subsets, the formation and possible functions of signaling molecular clusters that include ion channels in the immunological synapse, the discovery and refinement of structurally different ion channel blockers and the successful in vivo application of such compounds to suppress hypersensitivity reactions and autoimmune processes. In this review we wish to provide a concise update on these topics from recent years, highlighting the most notable developments. PMID- 20026118 TI - Evaluation of apoptotic and anti-apoptotic genes on efficacy of DNA vaccine encoding glycoprotein B of Herpes Simplex Virus type 1. AB - Many approaches have so far been tried to enhance the immunogenicity of DNA vaccine. These include the use of various factors that induce apoptosis or anti apoptosis effects when co-delivered with DNA vaccine. In the present study, the effects of pro-apoptotic Bax encoding plasmid (pBax) and anti-apoptotic Bcl-X(L) encoding plasmid (pBcl-xl), intradermally co-injected with glycoprotein B (gB) of Herpes Simplex Virus (HSV)-1 encoding plasmid (pgB) into the C57BL/6 mice were evaluated. Immune responses of the mice to the antigen were assessed by antibody assay, lymphoproliferative responses as well as cytokine and cytotoxic T lymphocyte (CTL) assay. Analysis of the humoral and cellular responses showed that the mice immunized with pBax and pgB induced higher levels of antibody and Interleukin-4 as well as stronger lymphocyte proliferative responses and cytotoxic activity compared to those mice received pgB alone. pBcl-xl when intradermally co-injected with pgB showed no significant enhancement in immune responses comparing to pgB. PMID- 20026120 TI - Development of a primer-probe energy transfer based real-time PCR for detection of Marek's disease virus. AB - A real-time PCR assay, which enables simultaneous detection and differentiation of all three serotypes of Marek's disease virus, without the need for post-PCR sequencing, has been developed. The assay is based on the primer-probe energy transfer real-time PCR, which has a relatively high tolerance towards point mutations in the probe region. The PCR is followed by a probe melting point analysis, which enables confirmation of identity of amplicon and differentiation of serotypes. The assay targets the MDV031 gene, encoding UL19 major capsid protein-like protein and was shown to be quantitative, with a detection limit below 10TCID(50)/ml starting material. This sensitivity is similar to the one obtained with traditional virus cultivation. However, the PCR method can provide a laboratory result within a day, while the virus cultivation method takes more than a week to perform. The new method will be useful for testing of avian live viral vaccines and screening for extraneous agents. PMID- 20026121 TI - Recombinant protein-based ELISA for detection and differentiation of antibodies against avian reovirus in vaccinated and non-vaccinated chickens. AB - Two nonstructural genes, sigmaNS and P17, of avian reovirus (ARV) were cloned into the expression plasmid vector PGEX4T-1. Expressed proteins for sigmaNS and P17 of avian reovirus were purified and used as antigens. Three indirect sigmaNS enzyme-linked immunosorbent assays (ELISAs), sigmaNS-ELISA, P17-ELISA and sigmaNS P17-ELISA were optimized and used as specific tests. Serum samples from reovirus infected and vaccinated SPF chickens were tested with the three ELISAs and an agar gel precipitin (AGP) method. ELISAs specific for sigmaNS, P17 and sigmaNS P17 were able to detect specific antibodies for avian reovirus in 88.9%, 61.1%, and 88.9% in infected samples, respectively, whereas the AGP detected 55.6% of the infected samples. The detection rates of ELISA specific antibodies for sigmaNS, P17 and sigmaNS-P17 on sera of vaccinated chickens were 6.7%, 0% and 6.7%. However, in comparison the AGP method detected 60.6% of antibodies in serum samples from vaccinated chickens. The results showed that the use of ELISAs specific for the nonstructural proteins might be able to distinguish between reovirus vaccinated and infected chickens. Further studies are in progress to validate these recombinant protein-based ELISAs under field conditions. PMID- 20026122 TI - Renilla luciferase-based quantitation of Potato virus A infection initiated with Agrobacterium infiltration of N. benthamiana leaves. AB - A quantitation method based on the sensitive detection of Renilla luciferase (Rluc) activity was developed and optimized for Potato virus A (PVA; genus Potyviridae) gene expression. This system is based on infections initiated by Agrobacterium infiltration and subsequent detection of the translation of PVA::Rluc RNA, which is enhanced by viral replication, first within the cells infected initially and later by translation and replication within new cells after spread of the virus. Firefly luciferase (Fluc) was used as an internal control to normalize the Rluc activity. An approximately 10-fold difference in the Rluc/Fluc activity ratio between a movement-deficient and a replication deficient mutant was observed starting from 48h post Agrobacterium infiltration (h.p.i.). The Rluc activity derived from wild type (wt) PVA increased significantly between 48 and 72h.p.i. and the Rluc/Fluc activity deviated clearly from that of the mutant viruses. Quantitation of the Rluc and Fluc mRNAs by semi quantitative RT-PCR indicated that increases and decreases in the Renillareniformis luciferase (rluc) mRNA levels coincided with changes in Rluc activity. However, a subtle increase in the mRNA level led to pronounced changes in Rluc activity. PVA CP accumulation was quantitated by enzyme-linked immunosorbent assay. The increase in Rluc activity correlated closely with virus accumulation. PMID- 20026119 TI - Estradiol: a key biological substrate mediating the response to cocaine in female rats. AB - A consistent finding in drug abuse research is that males and females show differences in their response to drugs of abuse. In women, increased plasma estradiol is associated with increased vulnerability to the psychostimulant and reinforcing effects of drugs of abuse. Our laboratory has focused on the role of estradiol in modulating the response to cocaine. We have seen that ovariectomy increases the locomotor response to a single cocaine injection, whereas estradiol exacerbates the locomotor response to repeated cocaine administration. Cocaine induced sensitization of brain activity, as measured by fMRI, is also dependent on plasma estradiol. Moreover, we observed that although all ovariectomized rats show conditioned place preference to cocaine, it is more robust in ovariectomized rats with estradiol. Opioid receptors are enriched in brain regions associated with pleasure and reward. We find that in females, the effectiveness of kappa opioid agonists in decreasing the locomotor response to repeated cocaine varies with plasma estradiol. We also find that estradiol regulates the density of mu opioid receptors in brains areas associated with reward. These data hint that in females, estradiol modulates the behavioral effects of cocaine by regulating mu and kappa opioid signaling in mesocorticolimbic brain structures. Identifying the mechanisms that mediate differences in vulnerability to drugs of abuse may lead to effective therapeutic strategies for the treatment and prevention of addiction and relapse. We encourage health practitioners treating persons addicted to drugs to consider gender differences in response to particular pharmacotherapies, as well the sex steroid milieu of the patient. PMID- 20026123 TI - Measurement of bluetongue virus binding to a mammalian cell surface receptor by an in situ immune fluorescent staining technique. AB - A quantifiable in situ immune fluorescent assay (IFA) was developed to measure bluetongue virus (BTV) binding to mammalian cells. The utility of the assay was demonstrated with both Chinese hamster ovary (CHO) and bovine pulmonary artery endothelial (CPAE) cells. Since heparin sulfate (HS) has been shown to function as a receptor for a number of viruses, its role as a receptor for BTV was evaluated with the in situ IFA. Binding of BTV to both CHO and CPAE cells was inhibited in a dose dependent manner by HS. In addition, HS deficient CHO cells showed greatly diminished binding of BTV when compared to the parental cell line. The IFA protocol will find application, as a non-isotopic, quantifiable technique, to study virus-cell receptor interactions. Information gained from such studies will expand our understanding of the early steps in virus replication. PMID- 20026124 TI - Identification of zoonotic Giardia genotypes in fish. AB - Apart from a single record in a shark, there have been no published studies conducted on Giardia genotypes in fish. The present study investigated the prevalence of Giardia in cultured fingerlings (n=227), wild freshwater (n=227) and wild marine/estuarine species (n=255) of fish in Western Australia by PCR amplification at the 18S rRNA, glutamate dehydrogenase (gdh), triose phosphate isomerase (tpi) and beta-giardin (bg) loci. Results revealed a low prevalence of Giardia, 3.8% (27/709), in fish hosts. The zoonotic Giardia species, Giardia duodenalis assemblages A, B as well as G. duodenalis assemblage E and Giardia microti were detected. The identification of zoonotic species of Giardia highlights the public health importance of investigating parasites within fish host species. PMID- 20026125 TI - Characterization of a diversity of tetraphyllidean and rhinebothriidean cestode larval types, with comments on host associations and life-cycles. AB - Life-cycles of marine tapeworms of the orders Tetraphyllidea and Rhinebothriidea are poorly known primarily because their larvae typically lack species level, taxonomically distinguishing adult characteristics and using morphology they can be identified to genus, family or order only. This large-scale study conducted in the northern Gulf of Mexico includes adult cestodes (25 species) collected from sharks and rays (Elasmobranchii, eight species) and larval cestodes (27 species) collected from teleosts (Neopterygii, 46 species), bivalves and gastropods (Mollusca, 24 species), and shrimps (Crustacea, five species), comprising a phylogenetically (75 species in three phyla, 14 orders and 46 families) and ecologically (e.g., benthic, epibenthic, pelagic, euryhaline, stenohaline) diverse array of hosts of larval cestode. Molecular biology and morphology informed larval identification and facilitated the circumscription of suites of morphological features representing distinct larval types (i.e., collective groups). A total of 198 specimens comprising adult and larval tetraphyllideans and rhinebothriideans assigned to 12 genera were characterized for the partial (D1-D3) lsrDNA gene and analyzed separately and in combination with data derived from species belonging to an additional 21 genera available from GenBank. Eight larval types were identified and matched to one or several genera of Tetraphyllidea or Rhinebothriidea; morphological variation within these larval types was also documented. In combination with published reports of unique larval morphologies, 15 larval types were established and a key to their larvae presented. Overall, teleosts figured prominently in the life-cycles of tetraphyllideans and rhinebothriideans. Intermediate host specificity at the level of cestode genus was euryxenous, but limited host records suggest that host specificity at the level of cestode species may be more strict. To our knowledge, this is the first published study that approaches the elucidation of marine tapeworm life-cycles by incorporating morphological, molecular biological and phylogenetic methods using specimens collected on a regional scale and from wild caught hosts from four metazoan phyla. PMID- 20026126 TI - GRANINS: thirty-five happy years in the granulosome world. Preface. PMID- 20026127 TI - Ca(2+)/calmodulin-dependent protein kinase II contributes to intracellular pH recovery from acidosis via Na(+)/H(+) exchanger activation. AB - The Na(+)/H(+) exchanger (NHE-1) plays a key role in pH(i) recovery from acidosis and is regulated by pH(i) and the ERK1/2-dependent phosphorylation pathway. Since acidosis increases the activity of Ca(2+)/calmodulin-dependent protein kinase II (CaMKII) in cardiac muscle, we examined whether CaMKII activates the exchanger by using pharmacological tools and highly specific genetic approaches. Adult rat cardiomyocytes, loaded with the pH(i) indicator SNARF-1/AM were subjected to different protocols of intracellular acidosis. The rate of pH(i) recovery from the acid load (dpH(i)/dt)-an index of NHE-1 activity in HEPES buffer or in NaHCO(3) buffer in the presence of inhibition of anion transporters-was significantly decreased by the CaMKII inhibitors KN-93 or AIP. pH(i) recovery from acidosis was faster in CaMKII-overexpressing myocytes than in overexpressing beta-galactosidase myocytes (dpH(i)/dt: 0.195+/-0.04 vs. 0.045+/-0.010 min(-)(1), respectively, n=8) and slower in myocytes from transgenic mice with chronic cardiac CaMKII inhibition (AC3-I) than in controls (AC3-C). Inhibition of CaMKII and/or ERK1/2 indicated that stimulation of NHE-1 by CaMKII was independent of and additive to the ERK1/2 cascade. In vitro studies with fusion proteins containing wild-type or mutated (Ser/Ala) versions of the C-terminal domain of NHE-1 indicate that CaMKII phosphorylates NHE-1 at residues other than the canonical phosphorylation sites for the kinase (Ser648, Ser703, and Ser796). These results provide new mechanistic insights and unequivocally demonstrate a role of the already multifunctional CaMKII on the regulation of the NHE-1 activity. They also prove clinically important in multiple disorders which, like ischemia/reperfusion injury or hypertrophy, are associated with increased NHE-1 and CaMKII. PMID- 20026128 TI - Excision of titin's cardiac PEVK spring element abolishes PKCalpha-induced increases in myocardial stiffness. AB - Protein kinase C-alpha (PKCalpha) was recently reported to increase myocardial stiffness, an effect that was proposed to be due to phosphorylation of two highly conserved sites (S11878 and S12022) within the proline-glutamic acid-valine lysine (PEVK) rich spring element of titin. To test this proposal we investigated the effect of PKCalpha on phosphorylation and passive stiffness in a mouse model lacking the titin exons that contain these two phosphorylation sites, the PEVK knockout (KO). We used skinned, gelsolin-extracted, left ventricular myocardium from wildtype and PEVK KO mice. Consistent with previous work we found that PKCalpha increased passive stiffness in the WT myocardium by 27+/-6%. Importantly, this effect was completely abolished in KO myocardium. In addition, increases in the elastic and viscous moduli at a wide range of frequencies (properties important in diastolic filling) following PKCalpha incubation (27+/ 3% and 20+/-4%, respectively) were also ablated in the KO. Back phosphorylation assays showed that titin phosphorylation following incubation with PKCalpha was significantly reduced by 36+/-12% in skinned PEVK KO myocardial tissues. The remaining phosphorylation in the KO suggests that PKCalpha sites exist in the titin molecule outside the PEVK region; these sites are not involved in increasing passive stiffness. Our results firmly support that the PEVK region of cardiac titin is phosphorylated by PKCalpha and that this increases passive tension. Thus, the PEVK spring element is the critical site of PKCalpha's involvement in passive myocardial stiffness. PMID- 20026129 TI - ADAM17-mediated shedding of the IL6R induces cleavage of the membrane stub by gamma-secretase. AB - Interleukin-6 (IL6) signals are mediated by classic and trans-signaling. In classic signaling, IL6 first binds to the membrane bound Interleukin-6 Receptor (IL6R) whereas in trans-signaling, IL6 acts via a soluble form of the IL6R. Trans signaling via the soluble IL6R (sIL6R) was linked to chronic inflammation and cancer. The release of the IL6R is mediated by the disintegrin and metalloproteinases ADAM10 and ADAM17. To analyze the fate of the C-terminal cleavage fragment after ectodomain shedding we fused the IL6R C-terminally to two Z-domains of Protein-A (2Z-tag) or to GFP. A specific C-terminal fragment of the IL6R protein could be detected after ADAM17-induced shedding. Using gamma secretase inhibitors and gene-deficient cells, we demonstrate that after ADAM17 mediated cleavage, the IL6R C-terminal fragment was cleaved by the gamma secretase at the plasma membrane. We were, however, not able to detect an IL6R intracellular domain. After gamma-secretase cleavage IL6R cell surface expression was lost and gamma-secretase cleavage product(s) of the IL6R were endocytosed. No GFP-fluorescence of a gamma-secretase-cleaved IL6R-GFP fusion protein was observed in the nucleus. We therefore hypothesize that a potential IL6R intracellular domain fragment is not involved in nuclear signaling but rapidly degraded. PMID- 20026130 TI - Intermonomer hydrogen bonds enhance GxxxG-driven dimerization of the BNIP3 transmembrane domain: roles for sequence context in helix-helix association in membranes. AB - We determined the sequence dependence of human BNIP3 transmembrane domain dimerization using the biological assay TOXCAT. Mutants in which intermonomer hydrogen bonds between Ser172 and His173 are abolished show moderate interaction, indicating that side-chain hydrogen bonds contribute to dimer stability but are not essential to dimerization. Mutants in which a GxxxG motif composed of Gly180 and Gly184 has been abolished show little or no interaction, demonstrating the critical nature of the GxxxG motif to BNIP3 dimerization. These findings show that side-chain hydrogen bonds can enhance the intrinsic dimerization of a GxxxG motif and that sequence context can control how hydrogen bonds influence helix helix interactions in membranes. The dimer interface mapped by TOXCAT mutagenesis agrees closely with the interfaces observed in the NMR structure and inferred from mutational analysis of dimerization on SDS-PAGE, showing that the native dimer structure is retained in detergents. We show that TOXCAT and SDS-PAGE give complementary and consistent information about BNIP3 transmembrane domain dimerization: TOXCAT is insensitive to mutations that have modest effects on self association in detergents but readily discriminates among mutations that completely disrupt detergent-resistant dimerization. The close agreement between conclusions reached from TOXCAT and SDS-PAGE data for BNIP3 suggests that accurate estimates of the relative effects of mutations on native-state protein protein interactions can be obtained even when the detergent environment is strongly disruptive. PMID- 20026131 TI - Role of the adenine ligand on the stabilization of the secondary and tertiary interactions in the adenine riboswitch. AB - Riboswitches are RNA-based genetic control elements that function via a conformational transition mechanism when a specific target molecule binds to its binding pocket. To facilitate an atomic detail interpretation of experimental investigations on the role of the adenine ligand on the conformational properties and kinetics of folding of the add adenine riboswitch, we performed molecular dynamics simulations in both the presence and the absence of the ligand. In the absence of ligand, structural deviations were observed in the J23 junction and the P1 stem. Destabilization of the P1 stem in the absence of ligand involves the loss of direct stabilizing interactions with the ligand, with additional contributions from the J23 junction region. The J23 junction of the riboswitch is found to be more flexible, and the tertiary contacts among the junction regions are altered in the absence of the adenine ligand; results suggest that the adenine ligand associates and dissociates from the riboswitch in the vicinity of J23. Good agreement was obtained with the experimental data with the results indicating dynamic behavior of the adenine ligand on the nanosecond time scale to be associated with the dynamic behavior of hydrogen bonding with the riboswitch. Results also predict that direct interactions of the adenine ligand with U74 of the riboswitch are not essential for stable binding although it is crucial for its recognition. The possibility of methodological artifacts and force-field inaccuracies impacting the present observations was checked by additional molecular dynamics simulations in the presence of 2,6-diaminopurine and in the crystal environment. PMID- 20026132 TI - Motif III in superfamily 2 "helicases" helps convert the binding energy of ATP into a high-affinity RNA binding site in the yeast DEAD-box protein Ded1. AB - Motif III in the putative helicases of superfamily 2 is highly conserved in both its sequence and its structural context. It typically consists of the sequence alcohol-alanine-alcohol (S/T-A-S/T). Historically, it was thought to link ATPase activity with a "helicase" strand displacement activity that disrupts RNA or DNA duplexes. DEAD-box proteins constitute the largest family of superfamily 2; they are RNA-dependent ATPases and ATP-dependent RNA binding proteins that, in some cases, are able to disrupt short RNA duplexes. We made mutations of motif III (S A-T) in the yeast DEAD-box protein Ded1 and analyzed in vivo phenotypes and in vitro properties. Moreover, we made a tertiary model of Ded1 based on the solved structure of Vasa. We used Ded1 because it has relatively high ATPase and RNA binding activities; it is able to displace moderately stable duplexes at a large excess of substrate. We find that the alanine and the threonine in the second and third positions of motif III are more important than the serine, but that mutations of all three residues have strong phenotypes. We purified the wild-type and various mutants expressed in Escherichia coli. We found that motif III mutations affect the RNA-dependent hydrolysis of ATP (k(cat)), but not the affinity for ATP (K(m)). Moreover, mutations alter and reduce the affinity for single-stranded RNA and subsequently reduce the ability to disrupt duplexes. We obtained intragenic suppressors of the S-A-C mutant that compensate for the mutation by enhancing the affinity for ATP and RNA. We conclude that motif III and the binding energy of gamma-PO(4) of ATP are used to coordinate motifs I, II, and VI and the two RecA-like domains to create a high-affinity single-stranded RNA binding site. It also may help activate the beta,gamma-phosphoanhydride bond of ATP. PMID- 20026133 TI - A Concealed Information Test with multimodal measurement. AB - A Concealed Information Test (CIT) investigates differential physiological responses to deed-related (probe) vs. irrelevant items. The present study focused on the detection of concealed information using simultaneous recordings of autonomic and brain electrical measures. As a secondary issue, verbal and pictorial presentations were compared with respect to their influence on the recorded measures. Thirty-one participants underwent a mock-crime scenario with a combined verbal and pictorial presentation of nine items. The subsequent CIT, designed with respect to event-related potential (ERP) measurement, used a 3-3.5s interstimulus interval. The item presentation modality, i.e. pictures or written words, was varied between subjects; no response was required from the participants. In addition to electroencephalogram (EEG), electrodermal activity (EDA), electrocardiogram (ECG), respiratory activity, and finger plethysmogram were recorded. A significant probe-vs.-irrelevant effect was found for each of the measures. Compared to sole ERP measurement, the combination of ERP and EDA yielded incremental information for detecting concealed information. Although, EDA per se did not reach the predictive value known from studies primarily designed for peripheral physiological measurement. Presentation modality neither influenced the detection accuracy for autonomic measures nor EEG measures; this underpins the equivalence of verbal and pictorial item presentation in a CIT, regardless of the physiological measures recorded. Future studies should further clarify whether the incremental validity observed in the present study reflects a differential sensitivity of ERP and EDA to different sub-processes in a CIT. PMID- 20026134 TI - The cellular basis of cell sorting kinetics. AB - Cell sorting is a dynamical cooperative phenomenon that is fundamental for tissue morphogenesis and tissue homeostasis. According to Steinberg's differential adhesion hypothesis, the structure of sorted cell aggregates is determined by physical characteristics of the respective tissues, the tissue surface tensions. Steinberg postulated that tissue surface tensions result from quantitative differences in intercellular adhesion. Several experiments in cell cultures as well as in developing organisms support this hypothesis. The question of how tissue surface tension might result from differential adhesion was addressed in some theoretical models. These models describe the cellular interdependence structure once the temporal evolution has stabilized. In general, these models are capable of reproducing sorted patterns. However, the model dynamics at the cellular scale are defined implicitly and are not well-justified. The precise mechanism describing how differential adhesion generates the observed sorting kinetics at the tissue level is still unclear. It is necessary to formulate the concepts of cell level kinetics explicitly. Only then it is possible to understand the temporal development at the cellular and tissue scales. Here we argue that individual cell mobility is reduced the more the cells stick to their neighbors. We translate this assumption into a precise mathematical model which belongs to the class of stochastic interacting particle systems. Analyzing this model, we are able to predict the emergent sorting behavior at the population level. We describe qualitatively the geometry of cell segregation depending on the intercellular adhesion parameters. Furthermore, we derive a functional relationship between intercellular adhesion and surface tension and highlight the role of cell mobility in the process of sorting. We show that the interaction between the cells and the boundary of a confining vessel has a major impact on the sorting geometry. PMID- 20026135 TI - Blockade of interleukin-6 signaling suppressed cochlear inflammatory response and improved hearing impairment in noise-damaged mice cochlea. AB - Hearing impairment can be the cause of serious socio-economic disadvantages. Recent studies have shown inflammatory responses in the inner ear co-occur with various damaging conditions including noise-induced hearing loss. We reported pro inflammatory cytokine interleukin-6 (IL-6) was induced in the cochlea 6h after noise exposure, but the pathophysiological implications of this are still obscure. To address this issue, we investigated the effects of IL-6 inhibition using the anti-IL-6 receptor antibody (MR16-1). Noise-exposed mice were treated with MR16-1 and evaluated. Improved hearing at 4kHz as measured by auditory brainstem response (ABR) was noted in noise-exposed mice treated with MR16-1. Histological analysis revealed the decrease in spiral ganglion neurons was ameliorated in the MR16-1-treated group, while no significant change was observed in the organ of Corti. Immunohistochemistry for Iba1 and CD45 demonstrated a remarkable reduction of activated cochlear macrophages in spiral ganglions compared to the control group when treated with MR16-1. Thus, MR16-1 had protective effects both functionally and pathologically for the noise-damaged cochlea primarily due to suppression of neuronal loss and presumably through alleviation of inflammatory responses. Anti-inflammatory cytokine therapy including IL-6 blockade would be a feasible novel therapeutic strategy for acute sensory neural hearing loss. PMID- 20026136 TI - Intravenous levetiracetam in the rat pilocarpine-induced status epilepticus model: behavioral, physiological and histological studies. AB - PURPOSE: Status epilepticus is a neurological emergency associated with neuronal injury, lasting behavioral disturbance, and a high rate of mortality. Intravenous levetiracetam (LEV), an anti-epileptic drug approved to treat partial seizures, has recently been introduced. We sought to determine the effect of LEV administered intravenously in a chemoconvulsant model of status epilepticus. METHODS: We examined the effect of intravenous LEV in the rat lithium-pilocarpine model of status epilepticus. Ten or 30 min after the onset of behavioral status epilepticus, animals were treated with LEV (200-1200 mg/kg i.v.) administered in a single bolus. Behavioral responses were recorded. Selected animals had continuous EEG recording before, during and after the administration of LEV. Some animals were sacrificed 24 h after the experiment and processed for histochemical assessment of neuronal injury. RESULTS: When administered 30 min after the onset of behavioral epileptic seizures, transient attenuation of ictal behavior was observed in animals treated with 800 mg/kg or more of LEV. The duration of behavioral attenuation increased sharply as the dose rose to 1000 mg/kg or higher, from a mean of 4-23.6 min. When administered 10 min after seizure onset, 400 mg/kg of LEV resulted in transient ictal behavioral attenuation, and higher doses caused relatively longer periods of attenuation. Pretreatment with LEV prior to pilocarpine also delayed the onset of seizures. EEG recordings, however, showed no significant attenuation of ictal discharge. By contrast, TUNEL staining demonstrated less neuronal injury in hippocampii and other limbic structures in animals that responded behaviorally to LEV. CONCLUSIONS: Intravenous administration of LEV in a chemoconvulsant model of status epilepticus results in attenuation of behavioral manifestations of seizure discharge and in reduction of neuronal injury but does not significantly alter ictal discharge recorded by EEG. PMID- 20026138 TI - Acute ethanol treatment prevents endocannabinoid-mediated long-lasting disinhibition of striatal output. AB - Recent research has suggested that the neuronal circuit adaptations elicited by drugs of abuse share common features with traditional learning models, and that drugs of abuse cause long-term changes in behavior by altering synaptic function and plasticity. Especially, the endocannabinoid (eCB) system appears to be involved in the neuronal circuitry regulating ethanol (EtOH) preference in rodent. The aim of this study was to evaluate if acute EtOH exposure could modulate eCB-mediated plasticity in the dorsolateral striatum. Our data show that EtOH (20-50 mM) prevents eCB-mediated long-lasting disinhibition (DLL) of striatal output induced by a single stimulation train delivered at 5 Hz for 60 s, and reduces long-term depression (LTD) induced by low-frequency stimulation at inhibitory synapses. Acute EtOH-treatment also prevents DLL induced by the L-type calcium channel activator 2,5-dimethyl-4-[2-(phenylmethyl)benzoyl]-1H-pyrrole-3 carboxylic acid methylester (FPL64176; 500 nM), or by the cannabinoid 1 receptor (CB(1)R) agonist WIN55,212-2 (300 nM), indicating that EtOH affects eCB-signaling at a stage that is downstream from eCB production and release. Importantly, high frequency stimulation, or a higher concentration of WIN55,212-2 (1 muM), induces EtOH-insensitive depression of striatal output, suggesting that EtOH affects CB(1)R-mediated signaling in a synapse-specific manner. Maintaining the balance between excitation and inhibition is vital for neuronal networks, and EtOH mediated modulation of eCB-signaling might thus affect the stability and the fine tuning of neuronal circuits in the striatum. Our data suggest that changes in eCB signaling could be involved in the physiological response to acute alcohol intoxication. PMID- 20026137 TI - Agonist-induced restoration of hippocampal neurogenesis and cognitive improvement in a model of cholinergic denervation. AB - Loss of basal forebrain cholinergic innervation of the hippocampus and severe neuronal loss within the hippocampal CA1 region are early hallmarks of Alzheimer's disease, and are strongly correlated with cognitive status. Various therapeutic approaches involve attempts to enhance neurotransmission or to provide some level of neuroprotection for remaining cells. An alternative approach may involve the generation of new cells to replace those lost in AD. Indeed, a simple shift in the balance between cell generation and cell loss may slow disease progression and possibly even reverse existing cognitive deficits. One potential neurogenic regulator might be acetylcholine, itself, which has been shown to play a critical role in hippocampal development. Here, we report the effects of various cholinergic compounds on indices of hippocampal neurogenesis, demonstrating a significant induction following pharmacological activation of muscarinic M1 receptors, located on hippocampal progenitors in the adult brain. This is the first report that a small-molecule agonist may induce neurogenesis in the hippocampal CA1 region. Furthermore, such treatment reversed deficits in markers of neurogenesis and spatial working memory triggered by cholinergic denervation in a rodent model. This study suggests the use of small molecule, receptor agonists may represent a novel means to trigger the restoration of specific neuronal populations lost to a variety of neurodegenerative disorders, such as Parkinson's, Alzheimer's, Huntington's and Amyotrophic Lateral Sclerosis. PMID- 20026139 TI - Food-related family lifestyle associated with fruit and vegetable consumption among young adolescents in Belgium Flanders and the Veneto Region of Italy. AB - The present paper examined the relationship between fruit and vegetable consumption and food-related family lifestyle (dinner with parents, family food rules, and television viewing behaviors) in a sample of 14,407 adolescents from Belgium Flanders (N=7,904) and the Veneto Region of Italy (N=6,503) using multilevel binary logistic regression analyses. The data are part of the Health Behavior in School-aged Children cross-sectional survey. Daily dinner with parents, having more strict family food rules, and low television viewing behaviors were associated with daily fruit and vegetable intake. Watching television during meals was not associated with the outcome variables. PMID- 20026140 TI - When seeing depends on knowing: adults with Autism Spectrum Conditions show diminished top-down processes in the visual perception of degraded faces but not degraded objects. AB - Behavioural, neuroimaging and neurophysiological approaches emphasise the active and constructive nature of visual perception, determined not solely by the environmental input, but modulated top-down by prior knowledge. For example, degraded images, which at first appear as meaningless 'blobs', can easily be recognized as, say, a face, after having seen the same image un-degraded. This conscious perception of the fragmented stimuli relies on top-down priming influences from systems involved in attention and mental imagery on the processing of stimulus attributes, and feature-binding [Dolan, R. J., Fink, G. R., Rolls, E., Booth, M., Holmes, A., Frackowiak, R. S. J., et al. (1997). How the brain learns to see objects and faces in an impoverished context. Nature, 389, 596-599]. In Autism Spectrum Conditions (ASC), face processing abnormalities are well-established, but top-down anomalies in various domains have also been shown. Thus, we tested two alternative hypotheses: (i) that people with ASC show overall reduced top-down modulation in visual perception, or (ii) that top-down anomalies affect specifically the perception of faces. Participants were presented with sets of three consecutive images: degraded images (of faces or objects), corresponding or non-corresponding grey-scale photographs, and the same degraded images again. In a passive viewing sequence we compared gaze times (an index of focal attention) on faces/objects vs. background before and after viewers had seen the undegraded photographs. In an active viewing sequence, we compared how many faces/objects were identified pre- and post-exposure. Behavioural and gaze tracking data showed significantly reduced effects of prior knowledge on the conscious perception of degraded faces, but not objects in the ASC group. Implications for future work on the underlying mechanisms, at the cognitive and neurofunctional levels, are discussed. PMID- 20026141 TI - Findings from animals concerning when interactions between perirhinal cortex, hippocampus and medial prefrontal cortex are necessary for recognition memory. AB - Loss of recognition memory is a prominent feature of the human classical amnesic syndrome. Recognition memory requires judgments concerning prior occurrence. Such judgments can be made in a variety of ways using different types of information such as the relative familiarity of individual objects or locations, or the location of a previously encountered object, or when an object was previously encountered. We review findings of selective ablation studies which demonstrate that the perirhinal cortex, hippocampus and medial prefrontal cortex are differently involved in recognition memory processes involving these different types of information. This review also presents data from a series of disconnection analyses, which test whether the perirhinal cortex, hippocampus and medial prefrontal cortex form components of an integrated system for these recognition memory processes. These analyses reveal that it is necessary for the perirhinal cortex, medial prefrontal cortex and the hippocampus to interact, forming an integrated network, in recognition memory involving judgment of whether an object has been previously encountered in a particular place (object in-place recognition memory) and in judging which of two objects was encountered longer ago (temporal order memory). In contrast, such interactions are not necessary when judgments are made concerning the prior occurrence of an individual item without positional information being necessary for the judgment (object memory) or concerning the prior occurrence of some item at a particular location without object information being necessary for the judgment (location memory). PMID- 20026142 TI - Vasoactive intestinal peptide (VIP) receptor expression in monocyte-derived macrophages from COPD patients. AB - Vasoactive intestinal peptide (VIP) is one of the most abundant molecules found in the respiratory tract. Due to its anti-inflammatory and bronchodilatatory properties, it has been proposed as a novel treatment for chronic obstructive pulmonary disease (COPD). The actions of VIP are mediated via three different G protein-coupled receptors (VPAC1, VPAC2 and PAC1) which are expressed in the respiratory tract and on immunocompetent cells including macrophages. Alveolar macrophages (AM) are key players in the pathogenesis of COPD and contribute to the severity and progression of the disease. While VPAC1 has been reported to be elevated in subepithelial cells in smokers with chronic bronchitis, little is known about VPAC expression of AM in COPD patients. AM from COPD patients show a strong VPAC1 expression which exceeds VPAC2. A similar receptor expression pattern was also observed in lipopolysaccharide (LPS)-activated monocyte-derived macrophages (MDM) from healthy volunteers and COPD patients. VIP has been shown to down-regulate interleukin 8 (IL-8) secretion significantly in MDM after LPS stimulation. The response to VIP was similar in MDM from COPD patients and healthy volunteers. Our results indicate that VPAC1 up-regulation in macrophages is a common mechanism in response to acute and chronic pro-inflammatory stimuli. Although VPAC1 up-regulation is dominant, both receptor subtypes are necessary for optimal anti-inflammatory signaling. The high VPAC1 expression in AM may reflect the chronic pro-inflammatory environment found in the lung of COPD patients. Treatment with VIP may help to decrease the chronic inflammation in the lung of COPD patients. PMID- 20026143 TI - A similar pattern of neuronal Fos activation in 10 brain regions following exposure to reward- or aversion-associated contextual cues in mice. AB - Relapse triggered by drug-paired cues is a major obstacle for successful treatment of drug abuse. Patterns of brain activation induced by drug-paired cues have been identified in human and animal models, but lack of specificity poses a serious problem for craving or relapse interpretations. The goal of this study was to compare brain responses to contextual cues paired with a rewarding versus an aversive stimulus in a mouse model to test the hypothesis that different patterns of brain activation can be detected. Mice were trained to associate a common environmental context with an intraperitoneal injection of saline, lithium chloride or cocaine. After measuring each animal for conditioned place preference or aversion, mice were re-exposed to the context (CS+ or CS-) in absence of the reinforcer to analyze patterns of Fos expression in 10 brain regions chosen from previous literature. Levels of Fos in the cingulate cortex, paraventricular thalamic nucleus, paraventricular hypothalamic nucleus, and dentate gyrus differed in CS+ versus CS- groups, but the direction of the differences was the same for both lithium chloride (LiCl) and cocaine reinforcers. In the cingulate cortex, Fos was positively correlated with degree of place preference for cocaine or aversion to LiCl whereas in the periaqueductal gray the relationship was positive for LiCl and negative for cocaine. Results confirm Fos responses to reward- or aversion-paired cues are similar but specificity is detectable. Future studies are needed to comprehensively establish neuroanatomical specificity in conditioned responses to drugs as compared to other reinforcers. PMID- 20026144 TI - Early sexual experience and stressful conditions affect the response of young ewes to the male. AB - Exposure of anoestrous ewes to rams induces oestrous cycles (the 'ram effect'). This response is poor in young-sexually naive females, possibly because they lack sexual experience and are sensitive to stress. Firstly, we assessed the conditions required for the acquisition of sexual experience. We exposed naive females at 5 or 9 months of age to rams and subsequently assessed their response at 1 year of age, to the ram effect. Unexpectedly, 2 weeks pre-exposure at 5 months and under some conditions at 9 months, inhibited the ram effect at 1 year of age, suggesting that early contact with rams was stressful and had long-term negative effects on reproduction. Consequently, we assessed the combined effect of stress and sexual experience on responses to the ram effect. At 9 months of age, we pre-exposed naive females using sexually active rams to achieve 'positive' (voluntary contact) or 'negative' (forced contact) experiences, and assessed responses to the ram effect at a year of age. In parallel, we subjected naive females to stressful conditions at the time of the male effect. During pre exposure, females with 'negative' contact exhibited more vigilance and stress cues and had higher levels of cortisol than 'positive' contact ewes. Regardless of pre-exposure or stress, the response to the ram effect at a year of age was inhibited. These experiments confirm that contact with sexual partners can be stressful to young ewes and highlight the need to consider their emotional state when conditioning them to the presence of rams and when managing breeding programs. PMID- 20026146 TI - NPAS2 deletion impairs responses to restricted feeding but not to metabolic challenges. AB - Neuronal PAS domain protein 2 (Npas2) is a clock gene expressed widely in brain and peripheral tissues. NPAS2 is responsive to cellular metabolic state and mutation of this gene impairs adaptation to restricted feeding schedules, suggesting that NPAS2 is required for effective control of a food-entrainable oscillator. However, an alternative possibility, that NPAS2 is required for detection of metabolic cues signaling energy deficiency or for arousal of appropriate behavioral responses to such cues, as not been directly examined. Therefore, we examined the effect of targeted disruption of Npas2 on responses to several acute and chronic metabolic challenges. We found that under normal light dark and ad libitum feeding conditions, Npas2 knockout (KO) mice did not differ from wild-type (WT) controls with respect to diurnal feeding or blood glucose levels, body weight or size or body composition. Furthermore, feeding responses to overnight food deprivation, insulin- or 2-deoxy-d-glucose (2DG)-induced glucoprivation, mercaptoacetate (MA)-induced blockade of fatty acid oxidation and cold exposure did not differ by genotype. However, KO mice lost more weight than WT during overnight food deprivation and when placed on a 4-h restricted feeding schedule, even though food intake did not differ between groups. Thus, it appears that NPAS2 is not required for detection of or behavioral responses to a variety of acute or chronic metabolic deficits, but is more likely to be involved in effective synchronization of feeding behavior with scheduled food availability. PMID- 20026145 TI - Post-oral infusion sites that support glucose-conditioned flavor preferences in rats. AB - Rats learn to prefer a flavored solution (CS+) paired with a gastrointestinal glucose infusion over an alternate flavor (CS-) paired with a non-caloric infusion. Prior work implicates a post-gastric site of glucose action, which is the focus of this study. In Exp. 1, male rats (8-10/group) were infused in the duodenum (ID), mid-jejunum (IJ), or distal ileum (II) with 8% glucose or water as they drank saccharin-sweetened CS+ and CS- solutions, respectively, in one-bottle 30-min sessions. Two-bottle tests (no infusions) were followed by a second train test cycle. By the second test, the ID and IJ groups preferred the CS+ (69%, 67%) to the CS- but the II group did not (48%). Satiation tests showed that ID and IJ infusions of glucose reduced intake of a palatable solution similarly, while II infusions were ineffective. In Exp. 2, rats (10/group) drank CS solutions in one bottle, 30-min sessions and were given 2-h ID or hepatic portal vein (HP) infusions. The CS+ and CS- were paired with 10 ml infusions of 10% glucose and 0.9% saline, respectively. Following 8 training sessions, the ID group preferred the CS+ (67%) to the CS- but the HP group did not (47%) in a two-bottle test. The similar CS+ preferences displayed by ID and IJ, but not II groups implicate the jejunum as a critical site for glucose-conditioned preferences. A pre-absorptive glucose action is indicated by the CS+ preference displayed by ID but not HP rats in Exp. 2. Our data were obtained with non-nutritive CS solutions. HP glucose infusions are reported to condition preferences for a flavored food that itself has pre- and post-absorptive actions. Thus, there may be multiple sites for glucose conditioning with the upper or mid-intestines being the first site of action. PMID- 20026147 TI - Neonatal DHT but not E2 speeds induction of sexual receptivity in the musk shrew. AB - Neural aromatization of testosterone (T) to estrogen during development is thought to be important for sexual differentiation of many altricial mammals. We evaluated the effects of neonatal injections of the non-aromatizable androgen dihydrotestosterone propionate (DHTP) and estradiol (E2) on the copulatory behavior of the female musk shrew, an altricial insectivore. Following adult ovariectomy and replacement T, animals were paired with a stimulus female for two 60-minute copulatory behavior tests. The latency to induce sexual receptivity (in the form of tail-wagging by the female), mount latency and total number of mounts were recorded in experimental females and in a group of untreated control males. While neither hormone treatment significantly affected mounting behavior, DHTP treated animals induced receptivity faster and with latencies not significantly different from intact males, suggesting that early non-aromatizable androgens can have masculinizing actions by either increasing sexual motivation or making the treated animal more attractive to the stimulus female. Reliance on androgenic rather than estrogenic metabolites for the differentiation of courtship behaviors conforms to the pattern seen more typically in primates than rodents. PMID- 20026148 TI - Safety evaluation of pectin-derived acidic oligosaccharides (pAOS): genotoxicity and sub-chronic studies. AB - Pectin-derived acidic oligosaccharides (pAOS) are non-digestible carbohydrates to be used in infant formulae and medical nutrition. To support its safety, the genotoxic potential of pAOS was evaluated. pAOS was not mutagenic in the Ames test. Positive results were obtained in the chromosome aberration test only at highly cytotoxic concentrations. The effects obtained in the mouse lymphoma test were equivocal; pAOS was not mutagenic in vivo. A sub-chronic dietary study, preceded by 4-week parental and in utero exposure phase, investigated general safety. Administration of pAOS did not affect parental health nor pup characteristics. No effects specific for acidic oligosaccharides were observed in the subsequent sub-chronic study. Slight diffuse hyperplasia of epithelial layer of the urinary bladder was noted to result from concurrently elevated urinary sodium, due to high sodium in pAOS, and elevated urinary pH. This phenomenon was confirmed in a mechanistic (sub-chronic) study. In contrast, in rats fed pAOS in combination with NH(4)Cl, an acidifying agent, the induced low urinary pH completely prevented the development of urothelial hyperplasia. Hyperplasia induced by this mechanism in rats is considered not relevant to man. Based on the current knowledge we consider pAOS safe for human consumption under its intended use. PMID- 20026149 TI - Synthesis and anti-glioma activity of 25(R)-spirostan-3beta,5alpha,6beta,19 tetrol. AB - Malignant gliomas are common and aggressive brain tumours in adults. The rapid proliferation and diffuse brain migration are the main obstacles to successful treatment. Here, we show 25(R)-spirostan-3beta,5alpha,6beta,19-tetrol, a polyhydroxy steroid, is capable of suppressing proliferation and migration of C6 malignant glioma cells in a concentration-dependent manner. The compound 25(R) spirostan-3beta,5alpha,6beta,19-tetrol was synthesised by seven steps starting from diosgenin in 8.55% overall yield. The structures of the synthetic compounds were characterised by infrared (IR), (1)H nuclear magnetic resonance (NMR), (13)C NMR spectra and EA. PMID- 20026150 TI - Pituitary volumes in hypochondriac patients. AB - To date, no study has examined the pituitary volumes in patients with hypochondriasis. In the present study, we evaluated pituitary volumes in patients with hypochondriasis and healthy controls. Twenty individuals with hypochondriasis (ten males, ten females), aged 20 to 48 years, and healthy controls were included into the study. The pituitary volumes were obtained. Volumetric measurements were made with T1-weighted coronal MRI images, with 2.4 mm-thick slices, at 1.5 T, and were done blindly. Volumetric measurements did not demonstrate group differences in the brain measurements, i.e., whole brain volume, white, and gray matter volumes (P>0.05). We found significantly smaller pituitary volumes of the whole group of hypochondriac patients compared to healthy controls (age and ICV as covariates). To conclude, the results from the current investigation suggest that hypochondriac patients had smaller pituitary volumes compared with healthy controls. This could be the keystone to a better understanding of the neurobiological basis of hypochondriasis. PMID- 20026151 TI - Striatal Akt/GSK3 signaling pathway in the development of L-Dopa-induced dyskinesias in MPTP monkeys. AB - L-Dopa treatment, the gold standard therapy for Parkinson's disease, is hampered by motor complications such as dyskinesias. Recently, impairment of striatal Akt/GSK3 signaling was proposed to play a role in the mechanisms implicated in development of L-Dopa-induced dyskinesias in a rodent model of Parkinson's disease. The present experiment investigated in 1-methyl-4-phenyl-1,2,3,6 tetrahydropyridine (MPTP) monkeys, the effects on Akt/GSK3 of chronic L-Dopa treatment inducing dyskinesias compared to L-Dopa with CI-1041 (NMDA receptor antagonist) or a low dose of cabergoline (dopamine D2 receptor agonist) preventing dyskinesias. The extensive dopamine denervation induced by MPTP was associated with a decrease by about half of phosphorylated Akt(Ser473) levels in posterior caudate nucleus, anterior and posterior putamen; smaller changes were observed for phosphorylated Akt(Thr308) levels that did not reach statistical significance. Dopamine depletion reduced phosphorylated GSK3beta(Ser9) levels, mainly in posterior putamen whereas pGSK3beta(Tyr216) and pGSK3alpha(Ser21) were unchanged. In posterior caudate nucleus, anterior and posterior putamen of dyskinetic L-Dopa-treated MPTP monkeys, pAkt(Ser473) and pGSK3beta(Ser9) were elevated whereas L-Dopa+cabergoline treated MPTP monkeys without dyskinesias had lower values in posterior striatum as vehicle-treated MPTP monkeys. In non dyskinetic MPTP monkeys treated with L-Dopa+CI-1041, putamen pAkt(Ser473) and pGSK3beta(Ser9) levels remained elevated as in dyskinetic monkeys while in posterior caudate nucleus, these levels were low as vehicle-treated and lower than L-Dopa treated MPTP monkeys. Extent of phosphorylation of Akt and GSK3beta in putamen correlated positively with dyskinesias scores of MPTP monkeys; these correlations were higher with dopaminergic drugs (L-Dopa, cabergoline) suggesting implication of additional mechanisms and/or signaling molecules in the NMDA antagonist antidyskinetic effect. In conclusion, our results showed that in MPTP monkeys, loss of striatal dopamine decreased Akt/GSK3 signaling and that increased phosphorylation of Akt and GSK3beta was associated with L-Dopa-induced dyskinesias. PMID- 20026152 TI - Activation of the Nrf2/ARE pathway via S-alkylation of cysteine 151 in the chemopreventive agent-sensor Keap1 protein by falcarindiol, a conjugated diacetylene compound. AB - Under basal conditions, the interaction of the cytosolic protein Kelch-like ECH associated protein 1 (Keap1) with the transcription factor nuclear factor-E2 related factor 2 (Nrf2) results in a low level of expression of cytoprotective genes whose promoter region contains the antioxidant response element (ARE). In response to oxidants and electrophiles, Nrf2 is stabilized and accumulates in the nucleus. The mechanism for this effect has been proposed to involve thiol dependent modulation of Keap1, leading to loss of its ability to negatively regulate Nrf2. We previously reported that falcarindiol (heptadeca-1,9(Z)-diene 4,6-diyne-3,8-diol), which occurs in Apiaceae and the closely related Araliaceae plants, causes nuclear accumulation of Nrf2 and induces ARE-regulated enzymes. Here, we report the mechanism of Nrf2 induction by falcarindiol. NMR analysis revealed that the conjugated diacetylene carbons of falcarindiol acted as electrophilic moieties to form adducts with a cysteine (Cys) thiol. In addition, using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry and circular dichroism spectroscopy, it was demonstrated that falcarindiol alkylated Cys residues in Keap1 and altered the Keap1 secondary structure. Transfection studies using the purified Keap1 protein, a luciferase reporter construct, and an Nrf2-expressing plasmid indicated that the intact Keap1 protein suppressed Nrf2-mediated ARE-luciferase activity. On the other hand, the falcarindiol-alkylated Keap1 protein did not suppress such activity. Treatment of HEK293 cells overexpressing Keap1 with falcarindiol generated a high molecular weight (HMW) form of Keap1. Furthermore, the Cys151 residue in Keap1 was found to be uniquely required for not only the formation of HMW Keap1 but also an increase in ARE-luciferase activity by falcarindiol. Our results demonstrate that falcarindiol having conjugated diacetylene carbons covalently modifies the Cys151 residue in Keap1 and that the inactivation of Keap1 by falcarindiol leads to activation of the Nrf2/ARE pathway. PMID- 20026153 TI - Evidence against a direct role for oxidative stress in cadmium-induced axial malformation in the chick embryo. AB - Cadmium (Cd) is a powerful inducer of oxidative stress. It also causes ventral body wall defects in chick embryos treated at Hamburger-Hamilton stages 16-17. By measuring malondialdehyde levels (TBARS method) and cotreating with antioxidants (tempol, ascorbate, and N-acetylcysteine), we sought to determine if oxidative stress were directly related to teratogenesis. We also investigated the expression of mRNAs for antioxidant enzymes superoxide dismutase (SOD) -1 and -2, catalase (CAT), and glutathione peroxidase (GPx). RT-PCR showed reductions in SOD 1, SOD-2, and CAT 1 hour after treatment with Cd. MDA levels increased 4 hours after Cd, and remained elevated 24 hours after treatment. Of the antioxidants, only N-acetylcysteine reduced MDA levels to control values. Nonetheless, no antioxidant could reduce embryo lethality or malformation rates. Furthermore, MDA levels 24 hours after treatment were identical in malformed and normal embryos exposed to Cd. Hence, we conclude that oxidative stress may not have a direct role in Cd teratogenesis. PMID- 20026154 TI - Embryotoxic effects of the marine biotoxin okadaic acid on murine embryonic stem cells. AB - Okadaic acid (OA), a marine toxin produced by dinoflagellates, can accumulate in various bivalve molluscs. In humans, consumption of OA induces acute toxic effects like diarrhoea, nausea, vomiting and abdominal pain. OA is a potent inhibitor of protein phosphatase 1 (PP1) and 2A (PP2A), enzymes that are known to be critical regulators of embryonic development. To determine the embryotoxic potential of OA, we performed two independent cellular in-vitro assays, both of which are applicable for the detection of teratogenic compounds: (i) the validated embryonic stem cell test (EST) based on the morphological analysis of beating cardiomyocytes in embryoid bodies and (ii) the F9 cell assay quantifying the induction of cell differentiation by measuring the emitted luminescence of a reporter gene. In the presence of OA, beating cardiomyocytes in the EST were inhibited and the reporter gene in transiently transfected F9 cells was activated. Furthermore, OA treatment led to rapid morphological changes including cell rounding, the loss of cell-cell contacts and changed electrical impedance as monitored in real time by the xCELLigence system. The two independent bioassays (EST and F9 cell test) detected OA as a potential embryotoxic compound, since OA influences the differentiation process of cultured murine embryonic cells. PMID- 20026155 TI - Intra-specific variation in venom of the African Puff Adder (Bitis arietans): Differential expression and activity of snake venom metalloproteinases (SVMPs). AB - Bitis arietans is considered one of the most medically significant snakes in Africa, primarily due to a combination of its extensive geographical distribution, common occurrence and highly potent haemorrhagic and cytotoxic venom. Our investigation has revealed a remarkable degree of intra-species variation between pooled venom samples from different geographical origins across sub-Saharan Africa and Arabia, and within a group of individual specimens from the same origin in Nigeria as determined by a combination of immunological, biochemical and proteomic assays. We demonstrate significant quantitative and qualitative differences between B. arietans venom in terms of protein expression, immunogenicity and activity of snake venom metalloproteinases (SVMPs); toxins with a primary role in the haemorrhagic and tissue-necrotic pathologies suffered by envenomed victims. Specifically, we have identified a processed PII SVMP that exhibits striking inter-specimen variability. PMID- 20026156 TI - Damage to some contractile and cytoskeleton proteins of the sarcomere in rat neonatal cardiomyocytes after exposure to pavetamine. AB - Pavetamine, a cationic polyamine, is a cardiotoxin that affects ruminants. The animals die of heart failure after a period of four to eight weeks following ingestion of the plants that contain pavetamine. This immunofluorescent study was undertaken in rat neonatal cardiomyocytes (RNCM) to label some of the contractile and cytoskeleton proteins after exposure to pavetamine for 48 h. Myosin and titin were degraded in the RNCM treated with pavetamine and the morphology of alpha actin was altered, when compared to the untreated cells, while those of beta tubulin seemed to be unaffected. F-actin was degraded, or even absent, in some of the treated cells. On an ultrastructural level, the sarcomeres were disorganized or disengaged from the Z-lines. Thus, all three contractile proteins of the rat heart were affected by pavetamine treatment, as well as the F-actin of the cytoskeleton. It is possible that these proteins are being degraded by proteases like the calpains and/or cathepsins. The consequence of pavetamine exposure is literally a "broken heart". PMID- 20026157 TI - Red ginseng extract protects against aflatoxin B1 and fumonisins-induced hepatic pre-cancerous lesions in rats. AB - The current study was conducted to evaluate the chemoprevention effects of ginseng extract (GE) against pre-cancerous lesions in female Sprague-Dawley rats treated with aflatoxin B1 (AFB1) and fumonisin (FB). Six experimental groups treated for 12 weeks and included: the control group; the GE alone-treated group (150 mg/kg b.w); the group treated orally with AFB1 (17 microg/kg b.w) during the first 2 weeks and fed FB-contaminated diet (250 mg/kg diet) during the 6th to 8th weeks; the group treated with GE during the mycotoxin protocol and continued till week 10; the group treated with GE 2 weeks before AFB1 administration and continued till the end of FB treatment and the group treated with GE for 4 weeks after the toxin protocol stopped. The sequential mycotoxins treatment induced significant changes in serum biochemical parameters accompanied by severe histological and histochemical changes of the liver tissue. Treatment with GE during, before or after the treatment with the mycotoxins improved all biochemical parameters and histological picture of the liver. Moreover, treatment with GE after the administration of the mycotoxins was found to be more effective. It could be concluded that GE has a protective effects as pre cancerous lesions and therapeutic effects as well. PMID- 20026158 TI - Effects of lycopene, synbiotic and their association on early biomarkers of rat colon carcinogenesis. AB - This study evaluated whether a synergy exists for the combined treatment with lycopene and synbiotic on early biomarkers of colon carcinogenesis. Male Wistar rats received a diet containing 300 mg/kg of lycopene and/or synbiotic (Bifidobacterium lactisplus oligofructose/inulin) or their combination 2 weeks before and during carcinogen treatment with 1,2-dimethylhydrazine (DMH). Twenty four hours after the last DMH application, the colons were processed for immunohistochemical analysis of proliferating cell nuclear antigen (PCNA), p53 protein, hematoxylin-eosin staining for apoptosis analysis and genotoxicity of fecal water by comet assay. Eight weeks after the last DMH application, the colons were analyzed for development of classical aberrant crypt foci (ACF) and mucin-negative ACF. Treatment with lycopene, synbiotic or their combination significantly increased apoptosis, reduced the PCNA and p53 labeling indexes and the development of classical ACF and mucin-negative ACF. Furthermore, a lower genotoxicity of fecal water was also detected in the groups treated with the chemopreventive agents. An additive/synergistic effect of the combined treatment with lycopene/synbiotic was observed only for the fecal water genotoxicity and mucin-negative ACF parameters. These results indicate that an additive/synergistic of the combination of chemopreventive agents on the initiation phase of colon carcinogenesis can be detected using selective early biomarkers. PMID- 20026159 TI - Evaluation of anti-oxidant activities and total phenolic content of Chromolaena odorata. AB - The aim of this study was to assess the in vitro potential of chloroform extract of Chromolaena odorata leaves. The DPPH activity of the extract (0.1-5 mg/ml) was increased in a dose dependent manner, which was found in the range of 23.48 91.61% as compared to ascorbic acid (33.69-94.10%). The IC50 values of chloroform extract in DPPH radical, hydroxyl radical, nitric oxide, ABTS radical were obtained to be 0.31, 0.43, 0.28 and 1.32 mg/ml, respectively. However, the IC50 values for the standard ascorbic acid were noted to be 0.24, 0.41, 0.23 and 1 mg/ml, respectively. Measurement of total phenolic content of the chloroform extract of C. odorata was achieved using Folin-Ciocalteau reagent containing 242.2 mg/g of phenolic content, which was found significantly higher when compared to reference standard gallic acid. The results obtained in this study clearly indicate that C. odorata has a significant potential to use as a natural anti-oxidant agent. PMID- 20026160 TI - Acetylcholinesterase, butyrylcholinesterase, and tyrosinase inhibition studies and antioxidant activities of 33 Scutellaria L. taxa from Turkey. AB - The members of Scutellaria L. (Lamiaceae) is known to be rich particularly in flavonoids and among them, S. baicalensis has been recorded to be used for memory enhancing purpose. Therefore, we initiated a study to screen the methanol extracts prepared from the aerial parts of 33 Turkish Scutellaria species for their acetylcholinesterase (AChE) and butyrylcholinesterase (BChE) inhibitory activities, which are the key enzymes taking place in pathogenesis of Alzheimer's disease. Besides, the methanol extracts were tested in vitro against another enzyme, tyrosinase, which is associated with melanin hyperpigmentation. 2,2 Diphenyl-1-picrylhydrazyl (DPPH) radical scavenger effect, ferrous ion-chelating ability, and ferric-reducing antioxidant power (FRAP) of the ethyl acetate and methanol extracts were also determined. AChE, BChE, and tyrosinase inhibition of the extracts were performed on ELISA microplate reader by spectrophotometric method. The extracts showed weak inhibition against AChE and BChE, while the best tyrosinase inhibition was caused by the methanol extract of S. brevibracteata subsp. subvelutina. The extracts had a very high DDPH radical scavenging effect and moderate antioxidant activity in ferrous ion-chelating and FRAP tests. PMID- 20026161 TI - Constituents of Corydalis heterocarpa and their anti-proliferative effects on human cancer cells. AB - Two new coumarins, 1 and 2, along with four known coumarins (3-6) have been isolated from Corydalis heterocarpa. On the basis of spectroscopic and chemical methods, compounds 1 and 2 were elucidated as (2'S,7'S)-O-2-methylbutanoyl columbianetin and (2'S)-columbianetin-3'-sulfate, respectively. The anti proliferative activity against human cancer cells of compounds 1-6 isolated from C. heterocarpa was evaluated using a MTT assay and by mRNA expression of several factors related to apoptosis. Among them, compound 2 exerted the more potent anti proliferative activity compared with the other compounds treated. The potent inhibitory effect of compound 2 was produced by induction of apoptosis through activating Bax, p53 and p21 expressions. PMID- 20026162 TI - Chromosome aberrations, micronucleus and sperm head abnormalities in mice treated with natamycin, [corrected] a food preservative. AB - Natamycin [corrected] is used as preservative in foods. The genotoxic effects of the food preservative natamycin [corrected] were evaluated using chromosome aberrations and micronucleus test in bone marrow cells and sperm head abnormality assays in mice. Blood samples were taken from mice and levels of total testosterone in serum were also determined. Natamycin [corrected] was intraperitoneally (ip) injected at 200, 400 and 800 mg/kg. Natamycin [corrected] did not induce chromosome aberrations but significantly increased the number of micronucleated polychromatic erythrocytes in bone marrow and sperm head abnormalities at all concentrations and treatment periods. It also decreased MI at all concentrations for 6, 12 and 24h treatment periods. Natamycin [corrected] decreased PCE/NCE ratio at all concentrations for 48h in female mice, for 24 and 48h treatment periods in male mice. At the 800 mg/kg concentration, natamycin [corrected] decreased PCE/NCE ratio for 24 and 72h in female mice. A dose dependent increase was observed in the percentage of sperm head abnormalities. The levels of serum testosterone decreased dose-dependently. The obtained results indicate that natamycin [corrected] is not clastogenic, but it is aneugenic in mice bone marrow and it is a potential germ cell mutagen in sperm cells. PMID- 20026163 TI - Evolution of domain combinations in protein kinases and its implications for functional diversity. AB - Protein kinases phosphorylating Ser/Thr/Tyr residues in several cellular proteins exert tight control over their biological functions. They constitute the largest protein family in most eukaryotic species. Protein kinases classified based on sequence similarity in their catalytic domains, cluster into subfamilies, which share gross functional properties. Many protein kinases are associated or tethered covalently to domains that serve as adapter or regulatory modules, aiding substrate recruitment, specificity, and also serve as scaffolds. Hence the modular organisation of the protein kinases serves as guidelines to their functional and molecular properties. Analysis of genomic repertoires of protein kinases in eukaryotes have revealed wide spectrum of domain organisation across various subfamilies of kinases. Occurrence of organism-specific novel domain combinations suggests functional diversity achieved by protein kinases in order to regulate variety of biological processes. In addition, domain architecture of protein kinases revealed existence of hybrid protein kinase subfamilies and their emerging roles in the signaling of eukaryotic organisms. In this review we discuss the repertoire of non-kinase domains tethered to multi-domain kinases in the metazoans. Similarities and differences in the domain architectures of protein kinases in these organisms indicate conserved and unique features that are critical to functional specialization. PMID- 20026164 TI - Nitric oxide mitigates apoptosis in human endothelial cells induced by 9,10 phenanthrenequinone: role of proteasomal function. AB - It has been widely recognized that nitric oxide (NO) suppresses oxidative damage of endothelial cell, but little is known about its pathophysiological role in apoptotic induction by 9,10-phenanthrenequinone (9,10-PQ), a major quinone component in diesel exhaust particles. Here, we have investigated the change in NO level in human aortic endothelial cells and the effect of NO in each step of apoptotic signaling initiated by 9,10-PQ. Treatment with 9,10-PQ evoked a bell shaped production of NO, which was presumably due to increase in an active form of endothelial NO synthase. Pretreatment with exogenous NO decreased the susceptibility of the cells to 9,10-PQ, and retrieved from apoptotic signaling (reactive oxygen species generation, glutathione depletion and caspase activation) induced during exposure to high concentrations of 9,10-PQ. In addition, inhibition of endogenous NO production augmented the toxicity of 9,10 PQ. Interestingly, the 9,10-PQ treatment resulted in marked decreases in the proteasomal activities, which were partially abrogated by NO and a cell-permeable cGMP analog. These results indicate that proteasomal dysfunction by oxidative stress participates in the 9,10-PQ-induced apoptotic signaling and is ameliorated by NO via a cGMP-dependent pathway, thereby suggesting the protective role of NO in vascular damage caused by 9,10-PQ. PMID- 20026165 TI - Effects of acute low-dose combined treatment with rimonabant and sibutramine on appetite and weight gain in rats. AB - In view of its potential advantages, drug polytherapy is currently attracting significant interest in the field of obesity research. In this context, concurrent manipulation of serotonergic and cannabinoid pathways in rodents has been found to reduce food and fluid intake in both an additive or synergistic manner. To further assess the value of this polytherapeutic approach, the current study examined the acute effects of low-dose combinations of the cannabinoid CB1 receptor antagonist/inverse agonist rimonabant (0.5 mg/kg) and the dual serotonin and noradrenaline-reuptake inhibitor sibutramine (0.125 and 0.25 mg/kg) in male rats. Ethological analysis was used to generate comprehensive behavioural profiles, including the behavioural satiety sequence (BSS). Findings confirmed that, although neither drug given alone significantly altered food intake, feeding behaviour or weight gain, rimonabant per se tended to reduce consumption and time spent feeding while significantly increasing scratching and grooming responses. However, none of these effects of the CB1 receptor antagonist/inverse agonist was significantly altered by the presence of either dose of sibutramine. In striking contrast to recent reports of acute low-dose interactions (enhanced appetite suppression and reduced side-effects) between rimonabant and naloxone, present results would not appear to support the clinical potential of rimonabant/sibutramine polytherapy for obesity. PMID- 20026166 TI - Paradoxical simultaneous occurrence of amphetamine-induced conditioned taste aversion and conditioned place preference with the same single drug injection: a new "pre- and post-association" experimental paradigm. AB - The paradoxical phenomenon of co-existing physically aversive and psychologically rewarding effects of drugs is a crucial issue for drug addiction. The present study employed a new experimental paradigm to test whether the rewarding and aversive properties of amphetamine (AMPH) can exist simultaneously. Rats were given a 15 min period of exposure to saccharin injected with 0.15M NaCl or 1.5mg/kg AMPH and then were confined to one compartment of a test box for 30 min. After three paired and unpaired cycles, the aversive and rewarding effects were assessed. A reduction in consumption of the paired flavored solution provided evidence of avoidance while preference for the AMPH injection context provided evidence of rewarding effects. The present findings demonstrate that the development of AMPH-induced rewarding and aversive effects depends on the particular behavioral conditions and support both the task-dependent drug effects hypothesis and the reward comparison hypothesis. The formation of associations with stimuli that comes before (pre) vs. after (post) the unconditioned stimulus and the role of the dopaminergic system in such associations are discussed. PMID- 20026167 TI - Lipoic acid blocks seizures induced by pilocarpine via increases in delta aminolevulinic dehydratase and Na+, K+-ATPase activity in rat brain. AB - In the present study we investigated the effects of lipoic acid (LA) on delta aminolevulinic dehydratase (delta-ALA-D) and Na(+), K(+)-ATPase activities in rat brain after seizures induction by pilocarpine. Wistar rats were treated with 0.9% saline (i.p., control group), lipoic acid (10mg/kg, i.p., LA group), pilocarpine (400mg/kg, i.p., pilocarpine group), or the combination of LA (10mg/kg, i.p.) with pilocarpine (400mg/kg, i.p.), 30 min before administration of LA (LA plus pilocarpine group). After the treatments all groups were observed for 1h. The enzyme activities (delta-ALA-D and Na(+), K(+)-ATPase) were measured using spectrophotometric methods, and the results were compared with that obtained from saline and pilocarpine-treated animals. Neuroprotective effects of LA against seizures were evaluated based on those enzyme activities. The pilocarpine group showed a reduction in delta-ALA-D and Na(+), K(+)-ATPase activities after seizures. In turn, LA plus pilocarpine abolished the appearance of seizures and reversed the decreased in delta-ALA-D and Na(+), K(+)-ATPase activities produced by seizures, when compared to the pilocarpine seizing group. The results from the present study demonstrate that preadministration of LA abolished seizure episodes induced by pilocarpine in rat, probably by increasing delta-ALA-D and Na(+), K(+) ATPase activities in rat brain during seizures. PMID- 20026168 TI - Isolation of an acidic phospholipase A2 from the venom of the snake Bothrops asper of Costa Rica: biochemical and toxicological characterization. AB - Phospholipases A(2) (PLA(2)) are major components of snake venoms, exerting a variety of relevant toxic actions such as neurotoxicity and myotoxicity, among others. Since the majority of toxic PLA(2)s are basic proteins, acidic isoforms and their possible roles in venoms are less understood. In this study, an acidic enzyme (BaspPLA(2)-II) was isolated from the venom of Bothrops asper (Pacific region of Costa Rica) and characterized. BaspPLA(2)-II is monomeric, with a mass of 14,212 +/- 6 Da and a pI of 4.9. Its complete sequence of 124 amino acids was deduced through cDNA and protein sequencing, showing that it belongs to the Asp49 group of catalytically active enzymes. In vivo and in vitro assays demonstrated that BaspPLA(2)-II, in contrast to the basic Asp49 counterparts present in the same venom, lacks myotoxic, cytotoxic, and anticoagulant activities. BaspPLA(2) II also differed from other acidic PLA(2)s described in Bothrops spp. venoms, as it did not show hypotensive and anti-platelet aggregation activities. Furthermore, this enzyme was not lethal to mice at intravenous doses up to 100 microg (5.9 microg/g), indicating its lack of neurotoxic activity. The only toxic effect recorded in vivo was a moderate induction of local edema. Therefore, the toxicological characteristics of BaspPLA(2)-II suggest that it does not play a key role in the pathophysiology of envenomings by B. asper, and that its purpose might be restricted to digestive functions. Immunochemical analyses using antibodies raised against BaspPLA(2)-II revealed that acidic and basic PLA(2)s form two different antigenic groups in B. asper venom. PMID- 20026169 TI - Stabilization of transcription factor Nrf2 by tBHQ prevents oxidative stress induced amyloid beta formation in NT2N neurons. AB - Alzheimer's disease (AD) a progressive neurodegenerative disorder of later life, is characterized by brain deposition of amyloid beta-protein (Abeta) plaques, accumulation of intracellular neurofibrillatory tangles, synaptic loss and neuronal cell death. There is significant evidence that oxidative stress is a critical event in the pathogenesis of AD. In the present study Abeta formation was induced in NT2N neurons, one of the most appropriate cell line models in AD. Our results indicate that oxidative stress resulting from the treatment of H(2)O(2)/FeSO(4) and/or 4-hydroxy-2-noenal (HNE) can be inhibited in the presence of tBHQ, a known inducer of nuclear factor-erythroid 2 related factor 2 (Nrf2) in NT2N neurons and can therefore be used to elucidate the relationship between oxidative stress, Abeta formation and Nrf2. The role of Nrf2 was confirmed using retinoic acid as an inhibitor of Nrf2. It provides the first documentation that tBHQ not only protects the neurons against cell death but also decreases amyloid beta formation. Moreover, the results indicate that oxidative stress fosters Abeta formation in NT2N neurons, creating a vicious neurodegenerative loop. PMID- 20026170 TI - Are effects from a brief multiple behavior intervention for college students sustained over time? AB - OBJECTIVE: This study examined whether 3-month outcomes of a brief image-based multiple behavior intervention on health habits and health-related quality of life of college students were sustained at 12-month follow-up without further intervention. METHODS: A randomized control trial was conducted with 303 undergraduates attending a public university in southeastern US. Participants were randomized to receive either a brief intervention or usual care control, with baseline, 3-month, and 12-month data collected during fall of 2007. RESULTS: A significant omnibus MANOVA interaction effect was found for health-related quality of life, p=0.01, with univariate interaction effects showing fewer days of poor spiritual health, social health, and restricted recent activity, p's<0.05, for those receiving the brief intervention. Significant group by time interaction effects were found for driving after drinking, p=0.04, and moderate exercise, p=0.04, in favor of the brief intervention. Effect sizes typically increased over time and were small except for moderate size effects for social health-related quality of life. CONCLUSION: This study found that 3-month outcomes from a brief image-based multiple behavior intervention for college students were partially sustained at 12-month follow-up. PMID- 20026171 TI - Neurobiology of infectious diseases: bringing them out of neglect. PMID- 20026172 TI - Bioenergetic pathways in tumor mitochondria as targets for cancer therapy and the importance of the ROS-induced apoptotic trigger. AB - Mitochondria are emerging as idealized targets for anti-cancer drugs. One reason for this is that although these organelles are inherent to all cells, drugs are being developed that selectively target the mitochondria of malignant cells without adversely affecting those of normal cells. Such anti-cancer drugs destabilize cancer cell mitochondria and these compounds are referred to as mitocans, classified into several groups according to their mode of action and the location or nature of their specific drug targets. Many mitocans selectively interfere with the bioenergetic functions of cancer cell mitochondria, causing major disruptions often associated with ensuing overloads in ROS production leading to the induction of the intrinsic apoptotic pathway. This in-depth review describes the bases for the bioenergetic differences found between normal and cancer cell mitochondria, focussing on those essential changes occurring during malignancy that clinically may provide the most effective targets for mitocan development. A common theme emerging is that mitochondrially mediated ROS activation as a trigger for apoptosis offers a powerful basis for cancer therapy. Continued research in this area is likely to identify increasing numbers of novel agents that should prove highly effective against a variety of cancers with preferential toxicity towards malignant tissue, circumventing tumor resistance to the other more established therapeutic anti-cancer approaches. PMID- 20026173 TI - Cell therapy for diabetes: stem cells, progenitors or beta-cell replication? AB - The cure for type 1 diabetes (T1D) will require either the replacement or regeneration of insulin-producing cells, together with measures that prevent their immune-mediated destruction. Experiments in rodent models have found that pancreatic stem cells, committed progenitors and replicating beta-cells can all contribute to insulin-producing cell regeneration. The cellular and molecular mechanisms of these cells, both in vitro and in vivo, have been investigated by us and by others. Furthermore, our surgical research laboratory has developed a unique in vivo chamber model of T1D, allowing the assessment of the behaviour of different sources of insulin-producing cells with a view to their potential use in cell-based therapies. PMID- 20026174 TI - Morphogenesis of the thyroid gland. AB - Congenital hypothyroidism is mainly due to structural defects of the thyroid gland, collectively known as thyroid dysgenesis. The two most prevalent forms of this condition are abnormal localization of differentiated thyroid tissue (thyroid ectopia) and total absence of the gland (athyreosis). The clinical picture of thyroid dysgenesis suggests that impaired specification, proliferation and survival of thyroid precursor cells and loss of concerted movement of these cells in a distinct spatiotemporal pattern are major causes of malformation. In normal development the thyroid primordium is first distinguished as a thickening of the anterior foregut endoderm at the base of the prospective tongue. Subsequently, this group of progenitors detaches from the endoderm, moves caudally and ultimately differentiates into hormone-producing units, the thyroid follicles, at a distant location from the site of specification. In higher vertebrates later stages of thyroid morphogenesis are characterized by shape remodeling into a bilobed organ and the integration of a second type of progenitors derived from the caudal-most pharyngeal pouches that will differentiate into C-cells. The present knowledge of thyroid developmental dynamics has emerged from embryonic studies mainly in chicken, mouse and more recently also in zebrafish. This review will highlight the key morphogenetic steps of thyroid organogenesis and pinpoint which crucial regulatory mechanisms are yet to be uncovered. Considering the co-incidence of thyroid dysgenesis and congenital heart malformations the possible interactions between thyroid and cardiovascular development will also be discussed. PMID- 20026175 TI - A pro-drug of the green tea polyphenol (-)-epigallocatechin-3-gallate (EGCG) prevents differentiated SH-SY5Y cells from toxicity induced by 6-hydroxydopamine. AB - Regular consumption of green tea benefits people in prevention from cardiovascular disorders, obesity as well as neurodegenerative diseases. (-) Epigallocatechin-3-gallate (EGCG) is regarded as the most biologically active catechin in green tea. However, the stability and bioavailability of EGCG are restricted. The purpose of the present study was to investigate whether a pro drug, a fully acetylated EGCG (pEGCG), could be more effective in neuroprotection in Parkinsonism mimic cellular model. Retinoic acid (RA)-differentiated neuroblastoma SH-SY5Y cells were pre-treated with different concentrations of EGCG and pEGCG for 30 min and followed by incubation of 25 microM 6 hydroxydopamine (6-OHDA) for 24h. We found that a broad dosage range of pEGCG (from 0.1 to 10 microM) could significantly reduce lactate dehydrogenase release. Likewise, 10 microM of pEGCG was effective in reducing caspase-3 activity, while EGCG at all concentrations tested in the model failed to attenuate caspase-3 activity induced by 6-OHDA. Furthermore, Western-blot analysis showed that Akt could be one of the specific signaling pathways stimulated by pEGCG in neuroprotection. It was demonstrated that 25 microM of 6-OHDA significantly suppressed the phosphorylation level of Akt. Only pEGCG at 10 microM markedly increased its phosphorylation level compared to 6-OHDA alone. Taken together, as pEGCG has higher stability and bioavailability for further investigation, it could be a potential neuroprotective agent and our current findings may offer certain clues for optimizing its application in future. PMID- 20026177 TI - Clinical availability of skin biopsy in the diagnosis of Parkinson's disease. AB - To determine whether skin biopsy is practically useful in the premortem diagnosis for Parkinson's disease (PD), we examined Lewy pathology in the skin of the chest wall and leg, obtained from 6-mm punch biopsies, using phosphorylated alpha synuclein antibody in 20 patients with clinically diagnosed PD. Abnormal accumulation of alpha-synuclein was found in the chest skin of two (10%) of 20 patients, but not in the leg. Although skin biopsy combined with a conventional immunohistochemistry for alpha-synuclein is not sufficient as a diagnostic tool, we could firstly demonstrate Lewy pathology in premortem tissue. The skin remains to be a promising tissue to be examined for the premortem diagnosis of PD. PMID- 20026176 TI - Mosquito immune defenses against Plasmodium infection. AB - The causative agent of malaria, Plasmodium, has to undergo complex developmental transitions and survive attacks from the mosquito's innate immune system to achieve transmission from one host to another through the vector. Here we discuss recent findings on the role of the mosquito's innate immune signaling pathways in preventing infection by the Plasmodium parasite, the identification and mechanistic description of novel anti-parasite molecules, the role that natural bacteria harbored in the mosquito midgut might play in this immune defense and the crucial parasite and vector molecules that mediate midgut infection. PMID- 20026178 TI - Decreases in rat brain aquaporin-4 expression following intracerebroventricular administration of an endothelin ET B receptor agonist. AB - Aquaporins (AQPs) comprise a family of water channel proteins, some of which are expressed in brain. Expressions of brain AQPs are altered after brain insults, such as ischemia and head trauma. However, little is known about the regulation of brain AQP expression. Endothelins (ETs), vasoconstrictor peptides, regulate several pathophysiological responses of damaged nerve tissues via ET(B) receptors. To show possible roles of ET(B) receptors in the regulation of brain AQP expression, the effects of intracerebroventricular administration of an ET(B) agonist were examined in rat brain. In the cerebrum, the copy numbers of AQP4 mRNAs were highest among AQP1, 3, 4, 5 and 9. Continuous administration of 500 pmol/day Ala(1,3,11,15)-ET-1, an ET(B) selective agonist, into rat brain for 7 days decreased the level of AQP4 mRNA in the cerebrum, but had no effect on AQP1, 3, 5 and 9 mRNA levels. The level of AQP4 protein in the cerebrum decreased by the administration of Ala(1,3,11,15)-ET-1. Immunohistochemical observations of Ala(1,3,11,15)-ET-1-infused rats showed that GFAP-positive astrocytes, but not neurons, activated microglia or brain capillary endothelial cells, had immunoreactivity for AQP4. These findings indicate that activation of brain ET(B) receptors causes a decrease in AQP4 expression, suggesting that ET down-regulates brain AQP4 via ET(B) receptors. PMID- 20026179 TI - Perspective taking modulates event-related potentials to perceived pain. AB - Recent event-related brain potential (ERP) study disentangled an early automatic component and a late top-down controlled component of neural activities to perceived pain of others. This study assessed the hypothesis that perspective taking modulates the top-down controlled component but not the automatic component of empathy for pain by recording ERPs from 24 subjects who performed pain judgments of pictures of hands in painful or non-painful situations from either self-perspective or other-perspective. We found that, relative to non painful stimuli, painful stimuli induced positive shifts of ERPs at frontal central electrodes as early as 160 ms after sensory stimulation and this effect lasted until 700 ms. The amplitudes of ERPs at 230-250 ms elicited by painful stimuli negatively correlated with both subjective ratings of others' pain and self-unpleasantness in both self-perspective and other-perspective conditions. Neural response to perceived pain over the central-parietal area was significantly reduced at 370-420 ms when performing the pain judgment task from other-perspective compared to self-perspective. The results suggest that shifting between self-perspectives and other-perspectives modulates the late controlled component but not the early automatic component of neural responses to perceived pain. PMID- 20026180 TI - Striatal dopamine in the response of brain and liver unsaturated and saturated free fatty acids following administration of C75 in CD-1 mice. AB - In order to clarify the mechanism of action of cerulenin analog, C75, known to suppress feeding behavior, food intake was measured in adult CD-1 male mice n=5 per group, treated i.p. with 10 and 20mg/kg of C75. Animals in both treatment groups had significantly lower 24h food consumption rate relative to the control group injected with vehicle. Striatal monoamine neurotransmitters and striatal as well as liver long chain free fatty acids concentrations were subsequently evaluated in another group treated i.p. with 20mg/kg C75. Acute exposure to C75 at 20mg/kg led to approximately 50% increase in the striatal dopamine levels and a decrease in dopamine turnover for up to 24h following the injection. The concentration of serotonin remained unchanged. Concentration of saturated fatty acids in the liver and striatum did not change, while striatal unsaturated myristoleic acid (cis-9-tetradecenoic acid) levels were significantly higher as early as 2h post-injection and remained elevated at 24h post-injection. These preliminary data suggest a central regulatory role of unsaturated fatty acids under dopaminergic control in the C75-induced anorexia. Pharmacological alterations in fatty acid metabolism may prove beneficial in the treatment of obesity. PMID- 20026181 TI - Disparate effects of serum on basal and evoked NFAT activity in primary astrocyte cultures. AB - In astrocytes, the Ca(2+)-dependent protein phosphatase calcineurin (CN) strongly regulates neuro-immune/inflammatory cascades through activation of the transcription factor, nuclear factor of activated T cells (NFAT). While primary cell cultures provide a useful model system for investigating astrocytic CN/NFAT signaling, variable results may arise both within and across labs because of differences in culture conditions. Here, we determined the extent to which serum and cell confluency affect basal and evoked astrocytic NFAT activity in primary cortical astrocyte cultures. Cells were grown to either approximately 50% or >90% confluency, pre-loaded with an NFAT-luciferase reporter construct, and maintained for 16 h in medium with or without 10% fetal bovine serum (FBS). NFAT-dependent luciferase expression was then measured 5h after treatment with vehicle alone to assess basal NFAT activity, or with Ca(2+) mobilizers and IL-1 beta to assess evoked activity. The results revealed significantly higher levels of basal NFAT activity in FBS-containing medium, regardless of cell confluency. Conversely, evoked NFAT activation was significantly lower in serum-containing medium, with an even greater inhibition observed in confluent cultures. Application of 10% FBS to serum-free astrocyte cultures quickly evoked a roughly seven-fold increase in NFAT activity that was significantly reduced by co-delivery of neutralizing agents for IL-1 beta, TNFalpha, and/or IFN gamma, suggesting that serum occludes evoked NFAT activation through a cytokine-based mechanism. Together, the results demonstrate that the presence of serum and cell confluency have a major impact on CN/NFAT signaling in primary astrocyte cultures and therefore must be taken into consideration when using this model system. PMID- 20026182 TI - Tapentadol, but not morphine, selectively inhibits disease-related thermal hyperalgesia in a mouse model of diabetic neuropathic pain. AB - Neuropathic pain in diabetic patients is a common distressing symptom and remains a challenge for analgesic treatment. Selective inhibition of pathological pain sensation without modification of normal sensory function is a primary aim of analgesic treatment in chronic neuropathic pain. Tapentadol is a novel analgesic with two modes of action, mu-opioid receptor (MOR) agonism and noradrenaline (NA) reuptake inhibition. Mice were rendered diabetic by means of streptozotocin, and neuropathic hyperalgesia was assessed in a 50 degrees C hot plate test. Normal nociception was determined in control mice. Tapentadol (0.1-1mg/kg i.v.) and morphine (0.1-3.16 mg/kg i.v.) dose-dependently attenuated heat-induced nociception in diabetic animals with full efficacy, reaching >80% at the highest doses tested. Tapentadol was more potent than morphine against heat hyperalgesia, with ED(50) (minimal effective dose) values of 0.32 (0.316) and 0.65 (1)mg/kg, respectively. Non-diabetic controls did not show significant anti-nociception with tapentadol up to the highest dose tested (1mg/kg). In contrast, 3.16 mg/kg morphine, the dose that resulted in full anti-hyperalgesic efficacy under diabetic conditions, produced significant anti-nociception in non-diabetic controls. Selective inhibition of disease-related hyperalgesia by tapentadol suggests a possible advantage in the treatment of chronic neuropathic pain when compared with classical opioids, such as morphine. It is hypothesized that this superior efficacy profile of tapentadol is due to simultaneous activation of MOR and inhibition of NA reuptake. PMID- 20026184 TI - MDMA ("ecstasy") impairs learning in the Morris Water Maze and reduces hippocampal LTP in young rats. AB - 3,4-Methylenedioxymethamphetamine (MDMA), an important recreational psychostimulant drug, was examined for its ability to alter visuo-spatial learning and synaptic plasticity. Young rats received MDMA (0.2 and 2mg/kg s.c.) twice per day for 6 days while their visuo-spatial learning was tested using the Morris Water Maze. After this, animals were sacrificed and LTP induced in hippocampal slices. Visuo-spatial learning was impaired and LTP reduced, both dose-dependently, without changes in serotonin levels or paired-pulse facilitation. We conclude that low, nontoxic doses of MDMA, applied during several days, slow learning by impairing postsynaptic plasticity. PMID- 20026183 TI - Chronic social defeat downregulates the 5-HT1A receptor but not Freud-1 or NUDR in the rat prefrontal cortex. AB - The serotonin 1A receptor (5-HT1A) and its associated transcriptional regulators, five prime repressor element under dual repression (Freud-1) and nuclear-deformed epidermal autoregulatory factor (NUDR/Deaf-1) have been previously found to be the repressors for 5-HT1A in the serotonergic raphe neurons, and are also altered in postmortem brains of individuals with major depressive disorder (MDD) and in rats exposed to chronic restraint stress. We sought to find out if rats exposed to chronic social defeat (CSD) stress also show altered expression of these genes. Adult male Wistar rats were exposed to CSD stress for four consecutive weeks following which they were sacrificed and gene expression assessed in the prefrontal cortex (PFC) by quantitative real-time polymerase chain reaction. While CSD had no significant effects on NUDR and Freud-1 mRNA levels, 5-HT1A mRNA levels were significantly downregulated in defeated animals. The data suggest that regulatory factors other than Freud-1 and NUDR may be involved in the regulation of 5-HT1A expression in PFC during CSD stress. Furthermore, decreased levels of 5-HT1A following social defeat in the PFC are consistent with human postmortem results for this receptor in major depression and demonstrate the possibility that this receptor is involved in the pathophysiology of depression and other stress related disorders. PMID- 20026185 TI - 6-Hz primed low-frequency rTMS to contralesional M1 in two cases with middle cerebral artery stroke. AB - This case study contrasted two subjects with stroke who received 6-Hz primed low frequency repetitive transcranial magnetic stimulation (rTMS) to the contralesional primary motor area (M1) to disinhibit ipsilesional M1. Functional magnetic resonance imaging (fMRI) showed that the intervention disrupted cortical activation at contralesional M1. Subject 1 showed decreased intracortical inhibition and increased intracortical facilitation following intervention during paired-pulse TMS testing of ipsilesional M1. Subject 2, whose precentral knob was totally obliterated and who did not show an ipsilesional motor evoked potential at pretest, still did not show any at posttest; however, her fMRI did show a large increase in peri-infarct zone cortical activation. Behavioral results were mixed, indicating the need for accompanying behavioral training to capitalize on the brain organization changes induced with rTMS. PMID- 20026186 TI - Linking cognitive aging to alterations in dopamine neurotransmitter functioning: recent data and future avenues. AB - Molecular-imaging studies of dopaminergic neurotransmission measure biomarkers of dopamine (DA), such as the DA transporter and D(1) and D(2) receptor densities in the living brain. These studies indicate that individual differences in DA functions are linked to cognitive performance irrespective of age, and serve as powerful mediators of age-related decline in executive functioning, episodic memory, and perceptual speed. This focused review targets several recent findings pertaining to these relationships. Specifically, we discuss novel evidence concerning (a) the role of DA in within-person cognitive variability; (b) age related differences in DA release during cognitive processing; (c) DA release following cognitive training in younger and older adults; and (d) the relationship between DA and task-induced functional brain activity. Based on these lines of empirical inquiry, we outline a series of avenues for future research on aging, DA, and cognition. PMID- 20026187 TI - Ferritins and iron storage in plants. AB - Iron is essential for both plant productivity and nutritional quality. Improving plant iron content was attempted through genetic engineering of plants overexpressing ferritins. However, both the roles of these proteins in the plant physiology, and the mechanisms involved in the regulation of their expression are largely unknown. Although the structure of ferritins is highly conserved between plants and animals, their cellular localization differ. Furthermore, regulation of ferritin gene expression in response to iron excess occurs at the transcriptional level in plants, in contrast to animals which regulate ferritin expression at the translational level. In this review, our knowledge of the specific features of plant ferritins is presented, at the level of their (i) structure/function relationships, (ii) cellular localization, and (iii) synthesis regulation during development and in response to various environmental cues. A special emphasis is given to their function in plant physiology, in particular concerning their respective roles in iron storage and in protection against oxidative stress. Indeed, the use of reverse genetics in Arabidopsis recently enabled to produce various knock-out ferritin mutants, revealing strong links between these proteins and protection against oxidative stress. In contrast, their putative iron storage function to furnish iron during various development processes is unlikely to be essential. Ferritins, by buffering iron, exert a fine tuning of the quantity of metal required for metabolic purposes, and help plants to cope with adverse situations, the deleterious effects of which would be amplified if no system had evolved to take care of free reactive iron. PMID- 20026188 TI - 3beta-taraxerol of Mangifera indica, a PI3K dependent dual activator of glucose transport and glycogen synthesis in 3T3-L1 adipocytes. AB - BACKGROUND: The present study focuses on identifying and developing an anti diabetic molecule from plant sources that would effectively combat insulin resistance through proper channeling of glucose metabolism involving glucose transport and storage. METHODS: Insulin-stimulated glucose uptake formed the basis for isolation of a bioactive molecule through column chromatography followed by its characterization using NMR and mass spectroscopic analysis. Mechanism of glucose transport and storage was evaluated based on the expression profiling of signaling molecules involved in the process. RESULTS: The study reports (i) the isolation of a bioactive compound 3beta-taraxerol from the ethyl acetate extract (EAE) of the leaves of Mangifera indica (ii) the bioactive compound exhibited insulin-stimulated glucose uptake through translocation and activation of the glucose transporter (GLUT4) in an IRTK and PI3K dependent fashion. (iii) the fate of glucose following insulin-stimulated glucose uptake was ascertained through glycogen synthesis assay that involved the activation of PKB and suppression of GSK3beta. GENERAL SIGNIFICANCE: This study demonstrates the dual activity of 3beta-taraxerol and the ethyl acetate extract of Mangifera indica as a glucose transport activator and stimulator of glycogen synthesis. 3beta-taraxerol can be validated as a potent candidate for managing the hyperglycemic state. PMID- 20026189 TI - Early identification of leptospirosis-associated pulmonary hemorrhage syndrome by use of a validated prediction model. AB - OBJECTIVE: To identify prediction factors for the development of leptospirosis associated pulmonary hemorrhage syndrome (LPHS). METHODS: We conducted a prospective cohort study. The study comprised of 203 patients, aged > or =14 years, admitted with complications of the severe form of leptospirosis at the Emilio Ribas Institute of Infectology (Sao Paulo, Brazil) between 1998 and 2004. Laboratory and demographic data were obtained and the severity of illness score and involvement of the lungs and others organs were determined. Logistic regression was performed to identify independent predictors of LPHS. A prospective validation cohort of 97 subjects with severe form of leptospirosis admitted at the same hospital between 2004 and 2006 was used to independently evaluate the predictive value of the model. RESULTS: The overall mortality rate was 7.9%. Multivariate logistic regression revealed that five factors were independently associated with the development of LPHS: serum potassium (mmol/L) (OR = 2.6; 95% CI = 1.1-5.9); serum creatinine (micromol/L) (OR = 1.2; 95% CI = 1.1-1.4); respiratory rate (breaths/min) (OR = 1.1; 95% CI = 1.1-1.2); presenting shock (OR = 69.9; 95% CI = 20.1-236.4), and Glasgow Coma Scale Score (GCS) < 15 (OR = 7.7; 95% CI = 1.3-23.0). We used these findings to calculate the risk of LPHS by the use of a spreadsheet. In the validation cohort, the equation classified correctly 92% of patients (Kappa statistic = 0.80). CONCLUSIONS: We developed and validated a multivariate model for predicting LPHS. This tool should prove useful in identifying LPHS patients, allowing earlier management and thereby reducing mortality. PMID- 20026190 TI - Glial fibrillary acidic protein-expressing neural progenitors give rise to immature neurons via early intermediate progenitors expressing both glial fibrillary acidic protein and neuronal markers in the adult hippocampus. AB - Adult neurogenesis occurs in the subgranular zone (SGZ) of the dentate gyrus, where primary neuronal progenitors that express glial fibrillary acidic protein (GFAP) develop into granule neurons. Here, we used transgenic mice with mouse GFAP promoter-controlled enhanced green fluorescent protein (mGFAP-EGFP Tg mice) to examine how astrocyte-like progenitors differentiate into neuron-committed progenitors. Bromodeoxyuridine (BrdU) analysis indicated that proliferating cells in the neurogenic SGZ transiently expressed EGFP and GFAP, and finally differentiated into cells positive for the neuronal marker, Hu (Hu+). Most proliferating EGFP+ cells showed expression of the stem cell marker, Sox2, and formed clusters of two to four cells containing GFAP+/EGFP+ and GFAP-/EGFP+ cells. No GFAP-/EGFP+ cells were detected in non-neurogenic regions, such as CA1 and CA3 of the pyramidal cell layer. Together with the assumption that exogeneous EGFP has a higher stability than that of endogenous GFAP in the degradation process, it is highly probable that the GFAP-/EGFP+ cells were daughter cells or immediate progeny derived from GFAP+/EGFP+ cells. The subpopulation of proliferating GFAP+/EGFP+ cells expressed proneural protein Mash1 and neuronal marker Hu, while the proliferating GFAP-/EGFP+ cells expressed additional immature neuronal markers, such as polysialic acid-neural cell adhesion molecule (PSA-NCAM) and doublecortin. Therefore, these results suggest that through a few cell divisions, GFAP+ progenitors give rise to neuronal progenitors via neuron committed early intermediate progenitors that express both GFAP and Hu (and/or Mash1). The findings of the present study also indicated that mGFAP-EGFP Tg mice are useful animals for identifying the daughter cells or immediate progeny derived from GFAP+ neural progenitors. PMID- 20026192 TI - Phytomedicines for Candida-associated denture stomatitis. AB - Phytomedicines are becoming more popular all over the world. Denture stomatitis (DS) presents as an inflammatory reaction in denture-bearing patients, under maxillary prosthesis with Candida albicans being the principal etiological agent. Many different methods of treatment for DS have been observed. The effects of the main medicinal plants claimed to be useful as antifungal agents in the treatment of DS are reviewed. Herbal remedies that have shown potential promise are mentioned, although much research is still required. PMID- 20026193 TI - A novel real-time PCR assay for determination of viral loads in person infected with hepatitis B virus. AB - A novel LUX (Light Upon eXtension) primer-based real-time PCR assay was developed and evaluated in this study, which was designed to provide a cost-effective, specific and highly sensitive method for viral load determination of hepatitis B virus (HBV). The assay employed an effective and rapid nucleic acid extraction system based on magnetic beads. To evaluate its efficacy, this new viral DNA preparation method was compared with QIAamp Blood Mini Kit and the results showed a good correlation (r=0.971; P<0.001). The performance of the LUX real-time assay was validated by testing serial dilutions of HBV plasmid DNA (5 to 5 x 10(8)copies/reaction) and a good linear relationship was obtained between the Ct values and the log(10) concentration of the HBV DNA. The assay possessed high sensitivity and the detection limit of this system was as few as 25 copies/ml of serum. A total of 91 positive serum samples were detected to evaluate further the assay and the high specificity was confirmed by melting curve analysis. This assay provides an ideal tool for monitoring the treatment efficacy and studying the relationship between HBV viral load and the stage of disease. PMID- 20026191 TI - How and when the fMRI BOLD signal relates to underlying neural activity: the danger in dissociation. AB - Functional magnetic resonance imaging (fMRI) has become the dominant means of measuring behavior-related neural activity in the human brain. Yet the relation between the blood oxygen-level dependent (BOLD) signal and underlying neural activity remains an open and actively researched question. A widely accepted model, established for sensory neo-cortex, suggests that the BOLD signal reflects peri-synaptic activity in the form of the local field potential rather than the spiking rate of individual neurons. Several recent experimental results, however, suggest situations in which BOLD, spiking, and the local field potential dissociate. Two different models are discussed, based on the literature reviewed to account for this dissociation, a circuitry-based and vascular-based explanation. Both models are found to account for existing data under some testing situations and in certain brain regions. Because both the vascular and local circuitry-based explanations challenge the BOLD-LFP coupling model, these models provide guidance in predicting when BOLD can be expected to reflect neural processing and when the underlying relation with BOLD may be more complex than a direct correspondence. PMID- 20026194 TI - Replication efficiency of rolling-circle replicon-based plasmids derived from porcine circovirus 2 in eukaryotic cells. AB - In this study, a method was developed to measure replication rates of rolling circle replicon-based plasmids in eukaryotic cells. This method is based on the discriminative quantitation of MboI-resistant, non-replicated input plasmids and DpnI-resistant, replicated plasmids. To do so, porcine circovirus type 2 (PCV2) replicon-based plasmids were constructed. These plasmids contained the PCV2 origin of replication, the PCV2 Rep promoter and the PCV2 Rep gene. The results show that the replication rate depends on the length of the PCV2 replicon-based plasmid and not on the respective position of the Rep promoter and the promoter of the gene of interest that encodes the enhanced green fluorescent protein (eGFP). In all cases, it was necessary to add the Rep gene encoded by a plasmid and cotransfected as a replication booster. This method can evaluate the replication potential of replicon-based plasmids quickly and is thereby a promising tool for the development of plasmids for vaccine purposes. PMID- 20026195 TI - Perceptual learning in an appetitive Pavlovian procedure: analysis of the effectiveness of the common element. AB - Non-reinforced preexposure to two stimuli often enhances discrimination between them. Analyses of this perceptual learning phenomenon have mainly focused on the role played by the distinctive stimulus features; this study examined the contribution of the non-distinctive common elements. A standard appetitive Pavlovian procedure was used. Rats received two different schedules of exposure- alternated or blocked--to two compound auditory stimuli, AX and BX. In Experiment 1 a generalization test to BX that followed conditioning to AX showed that animals responded less, and hence discriminated better, following alternated exposure, thus extending the generality of this perceptual learning effect to standard appetitive Pavlovian procedures. The degree to which the common element X was mediating this effect was explored in the next three experiments. Experiment 2 assessed the effectiveness of X following conditioning to AX. Experiment 3 explored X's effectiveness throughout extensive conditioning to X. Experiment 4 tested the ability of X to overshadow a novel stimulus Y. The results were consistent with the suggestion that alternated preexposure can reduce the relative effectiveness of the common element. PMID- 20026196 TI - How to spend a token? Trade-offs between food variety and food preference in tufted capuchin monkeys (Cebus apella). AB - Humans and non-human animals often choose among different alternatives by seeking variety. Here we assessed whether variety-seeking, i.e. the tendency to look for diversity in services and goods, occurs in capuchin monkeys--South-American primates which--as humans--are omnivorous and susceptible to food monotony. Capuchins chose between a Variety-token, that allowed to select one among 10 different foods (one more-preferred and nine less-preferred) and a Monotony token, that--upon exchange with the experimenter--either allowed to select one among 10 units of the same more-preferred food or gave access to one unit of the more-preferred food. To examine how food preference affects variety-seeking, in the B-condition we presented nine moderately preferred foods, whereas in the C condition we presented nine low-preferred foods. Overall, capuchins preferred the Variety-token over the Monotony-token and often selected one of the less preferred foods. These results suggest that variety-seeking is rooted in our evolutionary history, and that it satisfies the need of experiencing stimulation from the environment; at the ultimate level, variety-seeking may allow the organism to exploit novel foods and obtain a correct nutritional intake. Finally, variety-seeking could have contributed to the transition from barter to money in many human cultures. PMID- 20026197 TI - People's study time allocation and its relation to animal foraging. AB - In this article we suggest a relation between people's metacognitively guided study time allocation strategies and animal foraging. These two domains are similar insofar as people use specific metacognitive cues to assist their study time allocation just as other species use cues, such as scent marking. People decline to study items that they know they already know, just as other species use a win-shift strategy - avoiding already visited and depleted patches - in foraging. People selectively study the easiest as-yet-unlearned items first, before turning to more difficult items just as other species take the 'just right' size and challenge of prey-the so-called Goldilocks principle. People use a stop rule by which they give up on one item and turn to another when the returns diminish just as others species use a stop rule that guides shifting from one patch to another. The value that each item is assigned on the criterion test, if known during study, influences which items people choose to study and how long they study them just as knowledge of the nutritional or energy value of the food influences choices and perseverance in foraging. Finally, study time allocation strategies can differ in their effectiveness depending upon the expertise of the student just as some species forage close to optimally while others do not. PMID- 20026198 TI - A Kunitz-type proteinase inhibitor from the midgut of the ixodid tick, Haemaphysalis longicornis, and its endogenous target serine proteinase. AB - Although previous studies strongly suggested the involvement of serine proteases in blood digestion in the midgut of ticks, the regulating molecules of these proteinases are still unidentified. A novel Haemaphysalis longicornis Kunitz-type serine proteinase inhibitor with a single Kunitz-domain (HlMKI) has been identified and its co-localization with a midgut-derived serine proteinase (HlSP) within the epithelial cells has been demonstrated. Recombinant HlMKI inhibited the hydrolytic activity of HlSP, suggesting that HlMKI is a possible inhibitor of HlSP and may be part of a regulatory system of midgut serine proteinases. PMID- 20026199 TI - Analysis of nucleation kinetics of poorly water-soluble drugs in presence of ultrasound and hydroxypropyl methyl cellulose during antisolvent precipitation. AB - In this paper, nucleation kinetics of four poorly water-soluble drugs namely, itraconazole (ITZ), griseofulvin (GF), ibuprofen (IBP) and sulfamethoxazole (SFMZ) when precipitated by liquid antisolvent precipitation using water as antisolvent is examined in order to identify thermodynamic and kinetic process parameters as well as material properties that affect nucleation rate and hence, the particle size. The nucleation rates have been estimated for precipitation with and without ultrasound and hydroxypropyl methyl cellulose (HPMC). It is found that the nucleation rates increase significantly in presence of ultrasound and HPMC. Analysis of nucleation kinetics indicates that an increase in diffusivity due to ultrasound and a decrease in solid-liquid interfacial surface tension due to HPMC result in higher nucleation rates. Analysis also shows that reduction in interfacial surface tension due to HPMC is higher for a drug with lowest aqueous solubility (such as ITZ) as compared to drugs with higher aqueous solubility. It is also observed that it is easy to precipitate submicron particles of a drug with lowest aqueous solubility (such as ITZ) compared to drug molecules (such as SFMZ) with higher aqueous solubility in presence of HPMC. PMID- 20026200 TI - A skin permeability model of insulin in the presence of chemical penetration enhancer. AB - Enhancing transdermal delivery of insulin using chemical penetration enhancers (CPEs) has several advantages over other non-traditional methods; however, lack of suitable predictive models, make experimentation the only alternative for discovering new CPEs. To address this limitation, a quantitative structure property relationship (QSPR) model was developed, for predicting insulin permeation in the presence of CPEs. A virtual design algorithm that incorporates QSPR models for predicting CPE properties was used to identify 48 potential CPEs. Permeation experiments using Franz diffusion cells and resistance experiments were performed to quantify the effect of CPEs on insulin permeability and skin structure, respectively. Of the 48 CPEs, 35 were used for training and 13 were used for validation. In addition, 12 CPEs reported in literature were also included in the validation set. Differential evolution (DE) was coupled with artificial neural networks (ANNs) to develop the non-linear QSPR models. The six descriptor model had a 16% absolute average deviation (%AAD) in the training set and 4 misclassifications in the validation set. Five of the six descriptors were found to be statistically significant after sensitivity analyses. The results suggest, molecules with low dipoles that are capable of forming intermolecular bonds with skin lipid bi-layers show promise as effective insulin-specific CPEs. PMID- 20026201 TI - A characterisation study on the application of inverted lyotropic phases for subcutaneous drug release. AB - An experimental characterisation of lipid mixtures consisting of inverted hexagonal and inverted cubic phases composed of soybean phosphatidylcholine (SPC) and glycerol dioleate (GDO) was performed. The release of five chromophores of varying lipophilicity, used as model drugs, was investigated. Two experimental setups were applied: one based on maintaining sink condition, while a constant volume release medium was employed for the other. For neither setup, no correlation between the model drug lipophilicity and the polarity of the carrier matrix was found. However, the lipid phases showed a prolonged release, spanning weeks, of the model drugs, which exhibit lipophilicity values ranging by four orders of magnitude. PMID- 20026202 TI - Protein kinase A regulation of P2X(4) receptors: requirement for a specific motif in the C-terminus. AB - The P2X purinergic receptor sub-family of ligand-gated ion channels are subject to protein kinase modulation. We have previously demonstrated that P2X(4)R signaling can be positively regulated by increasing intracellular cAMP levels. The molecular mechanism underlying this effect was, however, unknown. The present study initially addressed whether protein kinase A (PKA) activation was required. Subsequently a mutational approach was utilized to determine which region of the receptor was required for this potentiation. In both DT-40 3KO and HEK-293 cells transiently expressing P2X(4)R, forskolin treatment enhanced ATP-mediated signaling. Specific PKA inhibitors prevented the forskolin-induced enhancement of ATP-mediated inward currents in P2X(4)R expressing HEK-293 cells. To define which region of the P2X(4)R was required for the potentiation, mutations were generated in the cytoplasmic C-terminal tail. It was determined that a limited region of the C-terminus, consisting of a non-canonical tyrosine based sorting motif, was required for the effects of PKA. Of note, this region does not harbor any recognizable PKA phosphorylation motifs, and no direct phosphorylation of P2X(4)R was detected, suggesting that PKA phosphorylation of an accessory protein interacts with the endocytosis motif in the C-terminus of the P2X(4)R. In support of this notion, using Total Internal Reflection Fluorescence Microscopy (TIRF)? P2X(4)-EGFP was shown to accumulate at/near the plasma membrane following forskolin treatment. In addition, disrupting the endocytosis machinery using a dominant-negative dynamin construct also prevented the PKA-mediated enhancement of ATP-stimulated Ca(2+) signals. Our results are consistent with a novel mechanism of P2XR regulation, whereby PKA activity, without directly phosphorylating P2X(4)R, markedly enhances ATP-stimulated P2X(4)R currents and hence cytosolic Ca(2+) signals. This may occur at least in part, by altering the trafficking of a population of P2X(4)R present at the plasma membrane. PMID- 20026203 TI - Defective repair of 5-hydroxy-2'-deoxycytidine in Cockayne syndrome cells and its complementation by Escherichia coli formamidopyrimidine DNA glycosylase and endonuclease III. AB - Repair of the oxidized purine 8-oxo-7,8-dihydro-2'-deoxyguanosine is inefficient in cells belonging to both complementation groups A and B of Cockayne syndrome (CS), a developmental and neurological disorder characterized by defective transcription-coupled repair. We show here that both CS-A and CS-B cells are also defective in the repair of 5-hydroxy-2'-deoxycytidine (5-OHdC), an oxidized pyrimidine with cytotoxic and mutagenic properties. The defect in the repair of oxidatively damaged DNA in CS cells thus extends to oxidized pyrimidines, indicating a general flaw in the repair of oxidized lesions in this syndrome. The defect could not be reproduced in in vitro repair experiments on oligonucleotide substrates, suggesting a role for both CS-A and CS-B proteins in chromatin remodeling during 5-OHdC repair. Expression of Escherichia coli formamidopyrimidine DNA glycosylase (FPG) or endonuclease III complemented the 5 OHdC repair deficiency. Hence, the expression of a single enzyme, FPG from E. coli, stably corrects the delayed removal of both oxidized purines and oxidized pyrimidines in CS cells. PMID- 20026204 TI - Dietary phenolic acids and ascorbic acid: Influence on acid-catalyzed nitrosative chemistry in the presence and absence of lipids. AB - Acid-catalyzed nitrosation and production of potentially carcinogenic nitrosative species is focused at the gastroesophageal junction, where salivary nitrite, derived from dietary nitrate, encounters the gastric juice. Ascorbic acid provides protection by converting nitrosative species to nitric oxide (NO). However, NO may diffuse into adjacent lipid, where it reacts with O(2) to re-form nitrosative species and N-nitrosocompounds (NOC). In this way, ascorbic acid promotes acid nitrosation. Using a novel benchtop model representing the gastroesophageal junction, this study aimed to clarify the action of a range of water-soluble antioxidants on the nitrosative mechanisms in the presence or absence of lipids. Caffeic, ferulic, gallic, or chlorogenic and ascorbic acids were added individually to simulated gastric juice containing secondary amines, with or without lipid. NO and O(2) levels were monitored by electrochemical detection. NOC were measured in both aqueous and lipid phases by gas chromatography-tandem mass spectrometry. In the absence of lipids, all antioxidants tested inhibited nitrosation, ranging from 35.9 + or - 7.4% with gallic acid to 93 + or - 0.6% with ferulic acid. In the presence of lipids, the impact of each antioxidant on nitrosation was inversely correlated with the levels of NO they generated (R(2) = 0.95, p<0.01): gallic, chlorogenic, and ascorbic acid promoted nitrosation, whereas ferulic and caffeic acids markedly inhibited nitrosation. PMID- 20026205 TI - Evaluation of quantitative PCR reference genes for gene expression studies in Tribolium castaneum after fungal challenge. AB - To investigate gene expression in Tribolium castaneum exposed to Beauveria bassiana, reference genes for qPCR were evaluated. Of these, beta-actin, alpha tubulin, and RPS6 were not stable. The most stable were ribosomal protein genes, RPS3, RPS18, and RPL13a. Syntaxin1, syntaxin6, and E-cadherin may be appropriate for some experimental systems. PMID- 20026206 TI - Methionine oxidation stabilizes non-toxic oligomers of alpha-synuclein through strengthening the auto-inhibitory intra-molecular long-range interactions. AB - Oxidative stress and aggregation of the presynaptic protein alpha-synuclein (alpha-Syn) are implied in the pathogenesis of Parkinson's disease and several other neurodegenerative diseases. Various posttranslational modifications, such as oxidation, nitration and truncation, have significant effects on the kinetics of alpha-Syn fibrillation in vitro. alpha-Syn is a typical natively unfolded protein, which possesses some residual structure. The existence of long-range intra-molecular interactions between the C-terminal tail (residues 120-140) and the central part of alpha-Syn (residues 30-100) was recently established (Bertoncini et al. (2005) Proc Natl Acad Sci U S A 102, 1430-1435). Since alpha Syn has four methionines, two of which (Met 1 and 5) are at the N-terminus and the other two (Met 116 and 127) are in the hydrophobic cluster at the C-terminus of protein, the perturbation of these residues via their oxidation represents a good model for studying the effect of long-range interaction on alpha-Syn fibril formation. In this paper we show that Met 1, 116, and 127 are more protected from the oxidation than Met 5 likely due to the residual structure in the natively unfolded alpha-Syn. In addition to the hydrophobic interactions between the C terminal hydrophobic cluster and hydrophobic central region of alpha-Syn, there are some long-range electrostatic interactions in this protein. Both of these interactions likely serve as auto-inhibitors of alpha-Syn fibrillation. Methionine oxidation affects both electrostatic and hydrophobic long-range interactions in alpha-Syn. Finally, oxidation of methionines by H2O2 greatly inhibited alpha-Syn fibrillation in vitro, leading to the formation of relatively stable oligomers, which are not toxic to dopaminergic and GABAergic neurons. PMID- 20026207 TI - Cis-regulatory elements affecting the Nanos gene promoter in the germline stem cells. AB - Drosophila Nanos gene plays an important role in stem cell maintenance and body patterning. With the purpose of understanding the cis-regulatory machinery involved in the transcription of the nanos gene in the germline stem cells, we examined its promoter fragment from +97 to -708 relative to the transcription start site and identified enhancer elements located between position -108 and +97. Experiments with transgenic flies revealed that the minimal promoter (from 108 to +20) is sufficient in the germline stem cells for the GFP expression in transgenic Drosophila. Moreover, the flag-tagged nanos protein blotting experiments revealed that a short promoter fragment plus some sequences of the nos 5'UTR spanning -108 to +97 could efficiently drive the expression of the flag tagged [Nos-mRNA-nos3'UTR] transgene in transgenic flies indicating that the cis regulatory elements located between positions -108 and +97 of the nanos promoter are sufficient to fully transcribe the nanos mRNA. Deletion of the identified cis acting sequences from the promoter rendered it non-functional as it could no longer transcribe the nanos mRNA in transgenic flies thus revealing the importance of these sequences for the transcription of the nanos gene. PMID- 20026208 TI - Tamavidin, a versatile affinity tag for protein purification and immobilization. AB - Tamavidin 2 is a fungal avidin-like protein that binds biotin with high affinity and is highly produced in soluble form in Escherichia coli. By contrast, widely used biotin-binding proteins avidin and streptavidin are rarely produced in soluble form in E. coli. In this study, we describe an efficient system for one step purification and immobilization of recombinant proteins using tamavidin 2 as an affinity tag. A bacterial sialyltransferase and soybean agglutinin were fused to tamavidin 2 and expressed in E. coli and tobacco BY-2 cells, respectively. High-level expressions of the fusion proteins were detected (80 mg l(-1)E. coli culture for bacterial sialyltransferase-tamavidin 2 and 2 mg l(-1) BY-2 cell culture for soybean agglutinin-tamavidin 2). To immobilize and purify the fusion proteins, biotinylated magnetic microbeads were incubated with the soluble extract from each recombinant host producing the fusion protein and then washed thoroughly. As the result, both fusion proteins were immobilized tightly on the microbeads without substantial loss of activity and simultaneously highly purified (90-95% purity) on the microbeads. Biotin with a longer linker contributed to higher affinity between the fusion protein and biotin. These results suggest that tamavidin fusion technology is a powerful tool for production, purification, and immobilization of recombinant proteins. PMID- 20026209 TI - A novel multivalent vaccine based on secretary antigen-delivery induces protective immunity against Vibrio anguillarum and Aeromonas hydrophila. AB - In our previous work, four secretory antigen-delivery systems based on different signal peptides (SPs) (SP(hlyA), SP(rtxA), SP(vah3) and SP(empA)) have been successfully established in attenuated Vibrio anguillarum. To investigate the potential application of the antigen-delivery systems in V. anguillarum multivalent vector vaccine, the glyceraldehyde-3-phosphate dehydrogenase (GAPDH) from pathogenic Aeromonas hydrophila, as a putative protective antigen, was fused to the four delivery systems, respectively, and introduced into attenuated V. anguillarum strain MVAV6203 to get four GAPDH-delivery strains in this work. Immunodetection of GAPDH indicated that among the four constructs, the SP(empA) mediated GAPDH-delivery strain AV(pGap-empA) showed the highest level of GAPDH expression and secretion, and was determined as the multivalent vaccine candidate. Further immune protection evaluation of AV(pGap-empA) in turbot (Scophtalmus maximus) demonstrated that the vaccine candidate could induce efficient protection against V. anguillarum and A. hydrophila, suggesting that this vector vaccine had great potential in serving as a valuable multivalent live vaccine. PMID- 20026210 TI - Seizures in the developing brain result in a long-lasting decrease in GABA(B) inhibitory postsynaptic currents in the rat hippocampus. AB - Whether seizures in the developing brain cause long-term changes in the mature brain has been debated. We tested the hypothesis that a model of early-life seizures, induced by systemic injection of a GABA(B) receptor antagonist CGP56999A in immature rats, decreased GABA(B) receptor-mediated inhibitory postsynaptic currents (IPSCs) in the hippocampus of adolescent rats. Whole-cell recordings were made in CA1 pyramidal cells and dentate gyrus (DG) granule cells in vitro, 30-45 days after the rats had seizures induced by CGP56999A (1-1.5 mg/kg i.p.) or control saline injection on postnatal day 15. GABA(B) receptor mediated IPSCs were reduced in DG neurons but not in CA1 neurons of early-life seizure rats as compared to controls. Additionally, hippocampal neurons of early life seizure rats, as compared to those in control rats, showed a more depolarized resting membrane potential in both CA1 and DG, and a larger input resistance but reduced spike frequency adaptation in DG neurons. In conclusion, early-life seizures result in a long-lasting reduction in GABA(B) receptor mediated transmission in DG principal neurons and depolarization in CA1 and DG principal neurons. These alterations are expected to increase seizure susceptibility in the adult brain. PMID- 20026211 TI - Behavioral effects of neuropeptides in rodent models of depression and anxiety. AB - In recent years, studies have advocated neuropeptide systems as modulators for the behavioral states found in mood disorders such as depression and anxiety disorders. Neuropeptides have been tested in traditional animal models and screening procedures that have been validated by known antidepressants and anxiolytics. However, it has become clear that although these tests are very useful, neuropeptides have distinct behavioral effects and dose-dependent characteristics, and therefore, use of these tests with neuropeptides must be done with an understanding of their unique characteristics. This review will focus on the behavioral actions of neuropeptides and their synthetic analogs, particularly in studies utilizing various preclinical tests of depression and anxiety. Specifically, the following neuropeptide systems will be reviewed: corticotropin-releasing factor (CRF), urocortin (Ucn), teneurin C-terminal associated peptide (TCAP), neuropeptide Y (NPY), arginine vasopressin (AVP), oxytocin, the Tyr-MIF-1 family, cholecystokinin (CCK), galanin, and substance P. These neuropeptide systems each have a unique role in the regulation of stress like behavior, and therefore provide intriguing therapeutic targets for mood disorder treatment. PMID- 20026212 TI - Entamoeba histolytica: a unicellular organism containing two active genes encoding for members of the TBP family. AB - Entamoeba histolytica is the protozoan parasite which causes human amoebiasis. In this parasite, few encoding genes for transcription factors have been cloned and characterized. The E. histolytica TATA-box binding protein (EhTBP) is the first basal transcription factor that has been studied. To continue with the identification of other members of the basal transcription machinery, we performed an in silico analysis of the E. histolytica genome and found three loci encoding for polypeptides with similarity to EhTBP. One locus has a 100% identity to the previously Ehtbp gene reported by our group. The second locus encodes for a 212 aa polypeptide that is 100% identical to residues 23-234 from EhTBP. The third one encodes for a 216 aa polypeptide of 24kDa that showed 42.6% identity and 73.7% similarity to EhTBP. This protein was named E. histolytica TBP-related factor 1 (EhTRF1). Ehtrf1 gene was expressed in bacteria and the purified 28kDa recombinant polypeptide showed the capacity to bind to TATTTAAA-box by electrophoretic mobility shift assays. K(D) values for rEhTBP and rEhTRF1 were (1.71+/-2.90)x10(-12)M and (1.12+/-0.160)x10(-11)M, respectively. Homology modeling of EhTRF1 and EhTBP revealed that, although they were very similar, they showed some differences on their surfaces. Thus, E. histolytica is a unicellular organism having two members of the TBP family. PMID- 20026213 TI - Enhanced expression of C/EBP homologous protein (CHOP) precedes degeneration of fibrocytes in the lateral wall after acute cochlear mitochondrial dysfunction induced by 3-nitropropionic acid. AB - We previously reported that treatment of the rat cochlea with a mitochondrial toxin, 3-nitropropionic acid (3-NP), causes temporary to permanent hearing loss depending on the amount of the drug. Furthermore, apoptosis of cochlear lateral wall fibrocytes, which are important for maintaining the endolymph, is a predominant pathological feature in this animal model. 3-NP is known to induce oxidative stress as well as neuronal apoptosis. C/EBP homologous protein gene (chop) is one of the marker genes induced during endoplasmic reticulum (ER) stress, and is also considered to be involved in apoptosis. To elucidate the molecular mechanism of cochlear fibrocyte apoptosis induced by 3-NP, we studied spatiotemporal expression of C/EBP homologous protein (CHOP) and other signaling molecules related to ER stress as well as the appearance of apoptotic cells in the cochlear lateral wall after 3-NP treatment. Quantitative real-time PCR revealed that chop and activating transcription factor 4 gene (atf-4) showed marked increase within 6h, whereas expression of other ER stress-responsive genes such as grp78 and grp94 did not change. Immunohistochemistry showed that 3-NP treatment caused up-regulation of CHOP, especially in type II and type IV fibrocytes, followed by the appearance of terminal deoxynucleotidyl transferase mediated dUTP nick end-labeling (TUNEL)-positive apoptotic cells in the same confined area. Thus, apoptosis of lateral wall fibrocytes induced by 3-NP is likely to be mediated by induction of CHOP. These results contribute clarification of pathological mechanism of cochlear fibrocytes and may lead to development of novel therapeutic strategy for hearing loss. PMID- 20026214 TI - Neuroprotective effects of resveratrol on ischemic injury mediated by modulating the release of neurotransmitter and neuromodulator in rats. AB - The present study was carried out to elucidate the neuroprotective effect and influence of resveratrol on the extracellular levels of neurotransmitter and neuromodulator during ischemia/reperfusion in rats. Male rats were divided into three groups: sham operation, ischemia treatment, and ischemia combined with resveratrol treatment (resveratrol-treated group, 30 mg/kg intraperitoneally for 7 days). Cerebral ischemia was induced by using the model of middle cerebral artery occlusion. The dialysates in hypothalamus were obtained by brain microdialysis technique. The levels of sixteen amino acids and amines in microdialysate were monitored by capillary electrophoresis analysis. This study shows that the ischemic infarcts were significantly reduced and neurological functions were improved in resveratrol-treated group compared to ischemia group. The analysis results demonstrate that chronic treatment with resveratrol remarkably reduced the release of excitatory neurotransmitter glutamate, aspartate and neuromodulator d-Serine during ischemia and reperfusion; and significantly increased the basal extracellular levels of inhibitory neurotransmitter gamma-amino-n-butyric acid, glycine and taurine. Chronic treatment with resveratrol also ameliorated O-phosphoethanolamine levels and excitotoxic index during ischemia and reperfusion. This study provides the first in vivo evidence that resveratrol could exert neuroprotective effect against ischemia injury by modulating the release of multiple neurotransmitters and neuromodulators during ischemia/reperfusion. PMID- 20026215 TI - Towards the understanding of resistance mechanisms in clinically isolated trimethoprim-resistant, methicillin-resistant Staphylococcus aureus dihydrofolate reductase. AB - Resistance to therapeutics such as trimethoprim-sulfamethoxazole has become an increasing problem in strains of methicillin-resistant Staphylococcus aureus (MRSA). Clinically isolated trimethoprim-resistant strains reveal a double mutation, H30N/F98Y, in dihydrofolate reductase (DHFR). In order to develop novel and effective therapeutics against these resistant strains, we evaluated a series of propargyl-linked antifolate lead compounds for inhibition of the mutant enzyme. For the propargyl-linked antifolates, the F98Y mutation generates minimal (between 1.2- and 6-fold) losses of affinity and the H30N mutation generates greater losses (between 2.4- and 48-fold). Conversely, trimethoprim affinity is largely diminished by the F98Y mutation (36-fold) and is not affected by the H30N mutation. In order to elucidate a mechanism of resistance, we determined a crystal structure of a complex of this double mutant with a lead propargyl-linked antifolate. This structure suggests a resistance mechanism consistent both for the propargyl-linked class of antifolates and for trimethoprim that is based on the loss of a conserved water-mediated hydrogen bond. PMID- 20026216 TI - Non-equilibrium silk fibroin adhesives. AB - Regenerated silkworm silk solutions formed metastable, soft-solid-like materials (e-gels) under weak electric fields, displaying interesting mechanical characteristics such as dynamic adhesion and strain stiffening. Raman spectroscopy, in situ electric field dynamic oscillatory rheology and polarized optical microscopy indicated that silk fibroin electrogelation involved intermolecular self-assembly of silk molecules into amorphous, micron-scale, micellar structures and the formation of relatively long lifetime, intermicellar entanglement crosslinks. Overall, the electrogelation process did not require significant intramolecular beta-strand or intermolecular beta-sheet formation, unlike silk hydrogels. The kinetics of e-gel formation could be tuned by changing the field strength and assembly conditions, such as silk concentration and solution pH, while e-gel stiffness was partially reversible by removal of the applied field. Transient adhesion testing indicated that the adhesive characteristics of e-gels could at least partially be attributed to a local increase in proton concentration around the positive electrode due to the applied field and surface effects. A working model of electrogelation was described en route to understanding the origins of the adhesive characteristics. PMID- 20026217 TI - Defining occupational and consumer exposure limits for enzyme protein respiratory allergens under REACH. AB - A wide range of substances have been recognized as sensitizing, either to the skin and/or to the respiratory tract. Many of these are useful materials, so to ensure that they can be used safely it is necessary to characterize the hazards and establish appropriate exposure limits. Under new EU legislation (REACH), there is a requirement to define a derived no effect level (DNEL). Where a DNEL cannot be established, e.g. for sensitizing substances, then a derived minimal effect level (DMEL) is recommended. For the bacterial and fungal enzymes which are well recognized respiratory sensitizers and have widespread use industrially as well as in a range of consumer products, a DMEL can be established by thorough retrospective review of occupational and consumer experience. In particular, setting the validated employee medical surveillance data against exposure records generated over an extended period of time is vital in informing the occupational DMEL. This experience shows that a long established limit of 60 ng/m(3) for pure enzyme protein has been a successful starting point for the definition of occupational health limits for sensitization in the detergent industry. Application to this of adjustment factors has limited sensitization induction, avoided any meaningful risk of the elicitation of symptoms with known enzymes and provided an appropriate level of security for new enzymes whose potency has not been fully characterized. For example, in the detergent industry, this has led to general use of occupational exposure limits 3-10 times lower than the 60 ng/m(3) starting point. In contrast, consumer exposure limits vary because the types of exposure themselves cover a wide range. The highest levels shown to be safe in use, 15 ng/m(3), are associated with laundry trigger sprays, but very much lower levels (e.g. 0.01 ng/m(3)) are commonly associated with other types of safe exposure. Consumer limits typically will lie between these values and depend on the actual exposure associated with product use. PMID- 20026218 TI - Understanding the relationships between spike rate and delta/gamma frequency bands of LFPs and EEGs using a local cortical network model. AB - Despite the widespread use of EEGs to measure the large-scale dynamics of the human brain, little is known on how the dynamics of EEGs relates to that of the underlying spike rates of cortical neurons. However, progress was made by recent neurophysiological experiments reporting that EEG delta-band phase and gamma-band amplitude reliably predict some complementary aspects of the time course of spikes of visual cortical neurons. To elucidate the mechanisms behind these findings, here we hypothesize that the EEG delta phase reflects shifts of local cortical excitability arising from slow fluctuations in the network input due to entrainment to sensory stimuli or to fluctuations in ongoing activity, and that the resulting local excitability fluctuations modulate both the spike rate and the engagement of excitatory-inhibitory loops producing gamma-band oscillations. We quantitatively tested these hypotheses by simulating a recurrent network of excitatory and inhibitory neurons stimulated with dynamic inputs presenting temporal regularities similar to that of thalamic responses during naturalistic visual stimulation and during spontaneous activity. The network model reproduced in detail the experimental relationships between spike rate and EEGs, and suggested that the complementariness of the prediction of spike rates obtained from EEG delta phase or gamma amplitude arises from nonlinearities in the engagement of excitatory-inhibitory loops and from temporal modulations in the amplitude of the network input, which respectively limit the predictability of spike rates from gamma amplitude or delta phase alone. The model suggested also ways to improve and extend current algorithms for online prediction of spike rates from EEGs. PMID- 20026219 TI - Comparison of characteristics between region-and voxel-based network analyses in resting-state fMRI data. AB - Small-world networks are a class of networks that exhibit efficient long-distance communication and tightly interconnected local neighborhoods. In recent years, functional and structural brain networks have been examined using network theory based methods, and consistently shown to have small-world properties. Moreover, some voxel-based brain networks exhibited properties of scale-free networks, a class of networks with mega-hubs. However, there are considerable inconsistencies across studies in the methods used and the results observed, particularly between region-based and voxel-based brain networks. We constructed functional brain networks at multiple resolutions using the same resting-state fMRI data, and compared various network metrics, degree distribution, and localization of nodes of interest. It was found that the networks with higher resolutions exhibited the properties of small-world networks more prominently. It was also found that voxel based networks were more robust against network fragmentation compared to region based networks. Although the degree distributions of all networks followed an exponentially truncated power law rather than true power law, the higher the resolution, the closer the distribution was to a power law. The voxel-based analyses also enhanced visualization of the results in the 3D brain space. It was found that nodes with high connectivity tended have high efficiency, a co localization of properties that was not as consistently observed in the region based networks. Our results demonstrate benefits of constructing the brain network at the finest scale the experiment will permit. PMID- 20026220 TI - Predictive models of autism spectrum disorder based on brain regional cortical thickness. AB - Autism spectrum disorder (ASD) is a neurodevelopmental disorder with a wide phenotypic range, often affecting personality and communication. Previous voxel based morphometry (VBM) studies of ASD have identified both gray- and white matter volume changes. However, the cerebral cortex is a 2-D sheet with a highly folded and curved geometry, which VBM cannot directly measure. Surface-based morphometry (SBM) has the advantage of being able to measure cortical surface features, such as thickness. The goals of this study were twofold: to construct diagnostic models for ASD, based on regional thickness measurements extracted from SBM, and to compare these models to diagnostic models based on volumetric morphometry. Our study included 22 subjects with ASD (mean age 9.2+/-2.1 years) and 16 volunteer controls (mean age 10.0+/-1.9 years). Using SBM, we obtained regional cortical thicknesses for 66 brain structures for each subject. In addition, we obtained volumes for the same 66 structures for these subjects. To generate diagnostic models, we employed four machine-learning techniques: support vector machines (SVMs), multilayer perceptrons (MLPs), functional trees (FTs), and logistic model trees (LMTs). We found that thickness-based diagnostic models were superior to those based on regional volumes. For thickness-based classification, LMT achieved the best classification performance, with accuracy=87%, area under the receiver operating characteristic (ROC) curve (AUC)=0.93, sensitivity=95%, and specificity=75%. For volume-based classification, LMT achieved the highest accuracy, with accuracy=74%, AUC=0.77, sensitivity=77%, and specificity=69%. The thickness-based diagnostic model generated by LMT included 7 structures. Relative to controls, children with ASD had decreased cortical thickness in the left and right pars triangularis, left medial orbitofrontal gyrus, left parahippocampal gyrus, and left frontal pole, and increased cortical thickness in the left caudal anterior cingulate and left precuneus. Overall, thickness-based classification outperformed volume-based classification across a variety of classification methods. PMID- 20026221 TI - The COMT Val108/158Met polymorphism and medial temporal lobe volumetry in patients with schizophrenia and healthy adults. AB - Abnormalities of the medial temporal lobe have been consistently demonstrated in schizophrenia. A common functional polymorphism, Val108/158Met, in the putative schizophrenia susceptibility gene, catechol-O-methyltransferase (COMT), has been shown to influence medial temporal lobe function. However, the effects of this polymorphism on volumes of medial temporal lobe structures, particularly in patients with schizophrenia, are less clear. Here we measured the effects of COMT Val108/158Met genotype on the volume of two regions within the medial temporal lobe, the amygdala and hippocampus, in patients with schizophrenia and healthy control subjects. We obtained MRI and genotype data for 98 schizophrenic patients and 114 matched controls. An automated atlas-based segmentation algorithm was used to generate volumetric measures of the amygdala and hippocampus. Regression analyses included COMT met allele load as an additive effect, and also controlled for age, intracranial volume, gender and acquisition site. Across patients and controls, each copy of the COMT met allele was associated on average with a 2.6% increase in right amygdala volume, a 3.8% increase in left amygdala volume and a 2.2% increase in right hippocampus volume. There were no effects of COMT genotype on volumes of the whole brain and prefrontal regions. Thus, the COMT Val108/158Met polymorphism was shown to influence medial temporal lobe volumes in a linear-additive manner, mirroring its effect on dopamine catabolism. Taken together with previous work, our data support a model in which lower COMT activity, and a resulting elevation in extracellular dopamine levels, stimulates growth of medial temporal lobe structures. PMID- 20026222 TI - Modulation by EEG features of BOLD responses to interictal epileptiform discharges. AB - INTRODUCTION: EEG-fMRI of interictal epileptiform discharges (IEDs) usually assumes a fixed hemodynamic response function (HRF). This study investigates HRF variability with respect to IED amplitude fluctuations using independent component analysis (ICA), with the goal of improving the specificity of EEG-fMRI analyses. METHODS: We selected EEG-fMRI data from 10 focal epilepsy patients with a good quality EEG. IED amplitudes were calculated in an average reference montage. The fMRI data were decomposed by ICA and a deconvolution method identified IED-related components by detecting time courses with a significant HRF time-locked to the IEDs (F-test, p<0.05). Individual HRF amplitudes were then calculated for each IED. Components with a significant HRF/IED amplitude correlation (Spearman test, p<0.05) were compared to the presumed epileptogenic focus and to results of a general linear model (GLM) analysis. RESULTS: In 7 patients, at least one IED-related component was concordant with the focus, but many IED-related components were at distant locations. When considering only components with a significant HRF/IED amplitude correlation, distant components could be discarded, significantly increasing the relative proportion of activated voxels in the focus (p=0.02). In the 3 patients without concordant IED-related components, no HRF/IED amplitude correlations were detected inside the brain. Integrating IED-related amplitudes in the GLM significantly improved fMRI signal modeling in the epileptogenic focus in 4 patients (p<0.05). CONCLUSION: Activations in the epileptogenic focus appear to show significant correlations between HRF and IED amplitudes, unlike distant responses. These correlations could be integrated in the analysis to increase the specificity of EEG-fMRI studies in epilepsy. PMID- 20026223 TI - Probability dependence and functional separation of the object-related and mismatch negativity event-related potential components. AB - The separation of concurrently presented sounds into distinct auditory objects is accompanied by the elicitation of the object-related negativity (ORN) component of the event-related potential (ERP). In the present study, participants were exposed to tone sequences containing not only concurrent (harmonicity), but also sequential cues (similarity to previous stimuli in the series) for auditory object formation. Although ORN was consistently elicited by two-object relative to one-object sounds, it was modulated by the presentation rate. With increasing probability of sound segregation, ORN amplitude decreased, and its scalp topography shifted from a bilateral towards a unilateral (left-hemispheric) activation pattern. Probability dependence of the ORN matches previous behavioral data. The differential sensitivity of the left- and right-hemispheric response to this manipulation may explain why an ORN probability effect was not obtained in previous electrophysiological studies. The specific functional role of ORN for concurrent sound perception was confirmed by dissociating it from the mismatch negativity (MMN) component as a correlate of processing sequential cues. ORN probability dependence is argued to be functionally relevant for the interaction of concurrent and sequential mechanisms of auditory scene analysis. PMID- 20026225 TI - The neural basis of agency: an fMRI study. AB - Agency, a feeling that the self is the cause of action, has a strong relationship to the processing of discrepancies between the predicted multi-sensory feedback from one's intended action and its actual outcome (hereafter, agency error). Although previous studies have explored the neural basis of agency by assessing the brain's response to agency error, the effects found are confounded by two types of error irrelevant to agency: a mismatch between different sensory inputs in general (sensory mismatch, SM error) and a basic response to any type of prediction error (oddball error). In this functional magnetic resonance imaging study, we identified the neural response specific to agency error by dissociating it from responses to SM and oddball errors. Subjects played a game in which they controlled an on-screen character. Neural responses to rare events of violated control and congruency between types of audio-visual feedback were compared to dissociate agency from SM error. In a separate session, subjects viewed repetitive motions of the character, and neural responses to rare events of unpredictable change in movement were identified as related to oddball error. Agency-error-specific activation was observed in the supplementary motor area (SMA), left cerebellum, right posterior parietal cortex (PPC), and right extrastriate body area (EBA). Oddball errors also activated areas near the PPC and EBA peaks. SM errors activated the pre-SMA and the right posterior superior temporal sulcus. Our results suggest that the SMA, cerebellum, and some parts of the PPC and EBA serve as the neural bases of agency. PMID- 20026224 TI - Spatiotemporal imaging of cortical activation during verb generation and picture naming. AB - One hundred and fifty years of neurolinguistic research has identified the key structures in the human brain that support language. However, neither the classic neuropsychological approaches introduced by Broca (1861) and Wernicke (1874), nor modern neuroimaging employing PET and fMRI has been able to delineate the temporal flow of language processing in the human brain. We recorded the electrocorticogram (ECoG) from indwelling electrodes over left hemisphere language cortices during two common language tasks, verb generation and picture naming. We observed that the very high frequencies of the ECoG (high-gamma, 70 160 Hz) track language processing with spatial and temporal precision. Serial progression of activations is seen at a larger timescale, showing distinct stages of perception, semantic association/selection, and speech production. Within the areas supporting each of these larger processing stages, parallel (or "incremental") processing is observed. In addition to the traditional posterior vs. anterior localization for speech perception vs. production, we provide novel evidence for the role of premotor cortex in speech perception and of Wernicke's and surrounding cortex in speech production. The data are discussed with regards to current leading models of speech perception and production, and a "dual ventral stream" hybrid of leading speech perception models is given. PMID- 20026226 TI - Using conditional maximization to determine hyperparameters in model-based fMRI. AB - In model-based analysis of fMRI data, a neural or cognitive mathematical model of behavior is used to predict changes in fMRI activity. The model predictions are often applied as a parametric modulation of the main stimulus effect within the context of the general linear model (GLM). Using a mathematical model has become an important method for connecting fMRI signals to behavior because the model represents how stimulus processing leads to behavior, and the parametric modulation represents a specific test about the profile of stimulus-related fMRI activity (for review and discussion, see O'Doherty et al., 2007). However, in some cases the parameters of the mathematical model may be under-determined because there is a range of values that equally well account for behavior, or perhaps an exploratory analysis is desired. Thus, in order to fully gauge the applicability of some mathematical model it would be useful to understand how fMRI analysis depends on those parameters. Here, a conditional maximization procedure is developed to search for parameter values in the mathematical model as hyperparameters in the GLM. Simulations and analysis with real fMRI data show that conditional maximization is an effective and simple procedure for estimating hyperparameters. General recommendations and caveats for using hyperparameters in model-based fMRI analysis are also presented. PMID- 20026227 TI - Dopamine transporter gene variation modulates activation of striatum in youth with ADHD. AB - Polymorphisms in the 3'UTR variable number tandem repeat (VNTR) of exon 15 of the dopamine transporter gene (DAT1) have been linked to attention-deficit hyperactivity disorder (ADHD); moreover, variability in DAT1 3'UTR genotype may contribute to both heterogeneity of the ADHD phenotype and differences in response to stimulant medications. The impact of this VNTR on neuronal function in individuals with ADHD remains unclear despite evidence that the polymorphisms influence dopamine transporter expression. Thus, we used event-related functional magnetic resonance imaging to examine the impact of DAT1 3'UTR genotype on brain activation during response inhibition in unmedicated children and adolescents with ADHD. Twenty-one youth with ADHD who were homozygous for the 10-repeat (10R) allele of the DAT1 3'UTR and 12 youth who were carriers of the 9-repeat (9R) allele were scanned while they performed a Go/No-Go task. Response inhibition was modeled by contrasting activation during correct No-Go trials versus correct Go trials. Participants who were homozygous for the DAT1 3'UTR 10R allele and those who had a single 9R allele did not differ on percent of trials with successful inhibition, which was the primary measure of inhibitory control. Yet, youth with the DAT1 3'UTR 10R/10R genotype had significantly greater inhibitory control related activation than those with one 9R allele in the left striatum, right dorsal premotor cortex, and bilaterally in the temporoparietal cortical junction. These findings provide preliminary evidence that neural activity related to inhibitory control may differ as a function of DAT1 3'UTR genotype in youth with ADHD. PMID- 20026228 TI - Temporary disruption of the rat blood-brain barrier with a monoclonal antibody: a novel method for dynamic manganese-enhanced MRI. AB - Manganese (Mn(2+)) has limited permeability through the blood-brain barrier (BBB). Opening the BBB such that a sufficient amount of Mn(2+) enters the extracellular space is a critical step for dynamic manganese-enhanced magnetic resonance imaging (ME-MRI) experiments. The traditional BBB opening method uses intracarotid hyperosmolar stress which results in suboptimal BBB opening, and practically is limited to nonsurvival experiments due to substantial surgical trauma. In the present ME-MRI study, we investigate the feasibility of opening the BBB with an antibody that targets the endothelial barrier antigen (EBA) specifically expressed by rat endothelial cells. Results demonstrate that intravenous infusion of the anti-EBA agent SMI-71 leads to BBB disruption of the whole brain as detected by ME-MRI and confirmed by Evans blue dye staining. Physiologically, injection of SMI-71 leads to a hypertensive response followed by a sustained hypotensive response in animals anesthetized with urethane alone. Incorporating isoflurane partially mitigated both pressor responses. In general, BBB disruption via intravenous infusion of SMI-71 is straightforward and obviates technical difficulties associated with intracarotid hyperosmolar stress, opening new possibilities for in vivo neuroimaging with ME-MRI. The data also suggest that ME-MRI may be used as an imaging method to assess BBB integrity complementary to the Evans blue dye method, a classical but highly invasive technique, permitting longitudinal assessment of the integrity of the BBB on the same animal. PMID- 20026229 TI - Cerebral asymmetries in 12-week-old C57Bl/6J mice measured by magnetic resonance imaging. AB - Asymmetries of multiple components of the rodent cerebrum have been described at various levels of organization. Yet, despite its ubiquitous nature, many confusing and sometimes contradictory reports regarding structural asymmetries in the rodent brain have been published. There is a need, therefore, for a whole brain imaging analysis technique for asymmetry studies that is both accurate, reproducible and robust. To this end, a comprehensive three-dimensional examination of differences in brain structure in an inbred mouse strain was undertaken. The goal of this study was thus to use high-resolution magnetic resonance imaging to assess structural asymmetries in the adult C57Bl/6J mouse brain. Fixed brain T2-weighted images of 20 male C57Bl/6J mice were acquired on a 7T scanner at 32 microm isotropic resolution. We used voxel-based analyses to examine structural asymmetries throughout the whole mouse brain. The striatum, medial-posterior regions of the thalamus, and motor, sensorimotor, and visual cortex were found to be asymmetrical. The most significant asymmetry was found in the hippocampus and, specifically, the dentate gyrus. In each case, the left region was larger than the right. No other regions of the mouse brain showed structural asymmetry. The results in the dentate gyrus were confirmed using stereology, revealing a correlation of r=0.61 between magnetic resonance and stereological measures. Hippocampal, along with cortical asymmetry, has been discussed repeatedly in the literature, yet a clear pattern of directionality, until this point, has not been described. The findings of asymmetry in the striatum and absence of asymmetry in the rest of the brain are novel and show the advantage of using the whole-brain three-dimensional techniques developed herein for assessing asymmetry. PMID- 20026230 TI - A Bayesian spatiotemporal model for very large data sets. AB - Functional MRI provides a unique perspective of neuronal organization; however, these data include many complex sources of spatiotemporal variability, which require spatial preprocessing and statistical analysis. For the latter, Bayesian models provide a promising alternative to classical inference, which uses results from Gaussian random field theory to assess the significance of spatially correlated statistic images. A Bayesian approach generalizes the application of these ideas in that (1) random fields are used to model all spatial parameters, not solely observation error, (2) their smoothness is optimized, and (3) a broader class of models can be compared. The main problem, however, is computational, due to the large number of voxels in a brain volume. Sampling methods are time-consuming; however, approximate inference using variational Bayes (VB) offers a principled and transparent way to specify assumptions necessary for computational tractability. Penny et al. (2005b) described such a scheme using a joint spatial prior and approximated the joint posterior density with one that factorized over voxels. However, a further computational bottleneck is encountered when evaluating the log model evidence used to compare models. This has lead to dividing a brain volume into slices and treating each independently. This amounts to approximating the spatial prior over a full volume with stacked 2D priors. That is, smoothness along the z-axis is not included in the model. Here we describe a VB scheme that approximates the zero mean joint spatial prior with a non-zero mean empirical prior that factors over voxels, thereby overcoming this problem. We do this by modifying the original VB algorithm of Penny et al. using the conditional form of a so-called conditional autoregressive (CAR) prior to update a marginal prior over voxels. We refer to this as a spatially-informed voxel-wise prior (SVP) and use them to spatially regularise general linear model (GLM) and autoregressive (AR) coefficients (over time to model serial correlations). This algorithm scales more favourably with the number of voxels providing a truly 3D spatiotemporal model over volumes containing tens of thousands of voxels. We compare the scaling of compute times with the number of voxels and performance with a joint prior applied to synthetic and single-subject data. PMID- 20026231 TI - Load effects in auditory selective attention: evidence for distinct facilitation and inhibition mechanisms. AB - It is unknown whether facilitation and inhibition of stimulus processing represent one or two mechanisms in auditory attention. We performed electrophysiological experiments in humans to address these two competing hypothesis. Participants performed an attention task under low or high memory load. Facilitation and inhibition were measured by recording electrophysiological responses to attended and ignored sounds and comparing them to responses to these same sounds when attention was considered to be equally distributed towards all sounds. We observed two late frontally distributed components: a negative one in response to attended sounds, and a positive one to ignored sounds. These two frontally distributed responses had distinct timing and scalp topographies and were differentially affected by memory load. Taken together these results provide evidence that attention-mediated top-down control reflects the activity of distinct facilitation and inhibition mechanisms. PMID- 20026232 TI - Fatigue and damage accumulation of veneer porcelain pressed on Y-TZP. AB - OBJECTIVES: This study compared the reliability and fracture patterns of zirconia cores veneered with pressable porcelain submitted to either axial or off-axis sliding contact fatigue. METHODS: Forty-two Y-TZP plates (12mm x 12mm x 0.5mm) veneered with pressable porcelain (12mm x 12mm x 1.2mm) and adhesively luted to water aged composite resin blocks (12mm x 12mm x 4mm) were stored in water at least 7 days prior to testing. Profiles for step-stress fatigue (ratio 3:2:1) were determined from single load to fracture tests (n=3). Fatigue loading was delivered on specimen either on axial (n=18) or off-axis 30 degrees angulation (n=18) to simulate posterior tooth cusp inclination creating a 0.7mm slide. Single load and fatigue tests utilized a 6.25mm diameter WC indenter. Specimens were inspected by means of polarized-light microscope and SEM. Use level probability Weibull curves were plotted with 2-sided 90% confidence bounds (CB) and reliability for missions of 50,000 cycles at 200N (90% CB) were calculated. RESULTS: The calculated Weibull Beta was 3.34 and 2.47 for axial and off-axis groups, respectively, indicating that fatigue accelerated failure in both loading modes. The reliability data for a mission of 50,000 cycles at 200N load with 90% CB indicates no difference between loading groups. Deep penetrating cone cracks reaching the core-veneer interface were observed in both groups. Partial cones due to the sliding component were observed along with the cone cracking for the off-axis group. No Y-TZP core fractures were observed. CONCLUSIONS: Reliability was not significantly different between axial and off-axis mouth-motion fatigued pressed over Y-TZP cores, but incorporation of sliding resulted in more aggressive damage on the veneer. PMID- 20026233 TI - Nociceptin and its metabolite attenuate U0126-induced memory impairment through a nociceptin opioid peptide (NOP) receptor-independent mechanism. AB - Nociceptin binds to nociceptin opioid peptide (NOP) receptors. We reported that although high doses of nociceptin impaired memory function and that these effects were mediated via NOP receptors, low doses of nociceptin attenuated the memory impairment, and these attenuating effects were not mediated via NOP receptors. Even very low doses of nociceptin were biologically active and suggested a certain binding site for this peptide, but the mechanism underlying this attenuating effect has not yet been elucidated. In the present study, we investigated the effect of an intrahippocampal injection (i.h.) of nociceptin on memory impairment induced by U0126, a MEK inhibitor, and Rp-cAMPS, a PKA inhibitor in a step-down type passive avoidance test. U0126 (2.63 nmol/mouse, i.h.) impaired memory formation and training-dependent phosphorylation of ERK2 in the hippocampus. Co-administration of nociceptin (10 fmol/mouse) significantly attenuated memory impairment, while it did not attenuate the inhibition of training-dependent phosphorylation of ERK2 induced by U0126. On the other hand, nociceptin did not attenuate memory impairment induced by Rp-cAMPS (0.448 nmol/mouse, i.h.). Nociceptin (1 fmol/mouse) also attenuated U0126 (5.26 nmol/mouse)-induced memory impairment in NOP receptor knockout mice. Nociceptin was reported to metabolize into fragments (1-13) and (14-17) in vivo, which showed pharmacological activities without affecting NOP receptors. Our findings showed that nociceptin (14-17) (1 fmol/mouse) also attenuated U0126-induced memory impairment, while nociceptin (1-13) (0.1-10 fmol/mouse) did not attenuate memory impairment. These results suggest a novel action site or mechanism for the attenuating effects of nociceptin and its metabolite, and the sequence of nociceptin (14-17) is a critical structure. PMID- 20026234 TI - Mechanisms of meningeal invasion by a bacterial extracellular pathogen, the example of Neisseria meningitidis. AB - The blood-cerebrospinal fluid (CSF) barrier physiologically protects the meningeal spaces from bloodborne bacterial pathogens, due to the existence of specialized junctional interendothelial complexes. A few bacterial pathogens are able to reach the subarachnoidal space and cause bacterial meningitis in humans, a rare but dreadful disease. Surprisingly, most of them are extracellular commensals of the nasopharynx (Neisseria meningitidis, Streptococcus pneumoniae and Haemophilus influenzae) or of the digestive tract (Escherichia coli and Streptococcus agalactiae). The particular ability of these pathogens to induce meningitis is related to virulence factors that allow them to escape host innate immunity, to multiply within the serum, and to interact closely with the endothelial front line of defense of the blood-CSF barrier. In vitro studies using microvascular brain endothelial cell lines have shown that induced transcytosis may be a common route used by H. influenzae, S. pneumoniae, E. coli and S. agalactiae to reach the CSF. N. meningitidis is a strict human pathogen that interacts very tightly with endothelial cells. Adhesion of the meningococcus is mediated by type IV pili that induce a localized remodeling of the sub cortical cytoskeleton, leading to the formation of endothelial membrane protrusions that anchor bacterial colonies at the endoluminal face of the endothelial cell membrane, allowing a better resistance to blood flow. Recent work has shown that N. meningitidis is also able to recruit the polarity complex Par3/Par6/aPKC that re-routes endothelial cell adhesion molecules of interendothelial junctions, opening a paracellular route for bacteria to cross the endothelial barrier. PMID- 20026235 TI - Editorial: Resistance to antifungal agents. PMID- 20026236 TI - A cryptic role of a glycolytic-gluconeogenic enzyme (aldolase) in amino acid transporter turnover in Aspergillus nidulans. AB - In Aspergillus nidulans the fbaA1013 mutation results in reduced or total loss of growth on glycolytic and gluconeogenic carbon sources, respectively. It also negatively affects growth on several amino acids (including L-proline, L glutamate or L-aspartate) that the fungus can use as nitrogen source on glycolytic carbon sources. Complementation of the fbaA1013 mutation using an A. nidulans genomic library resulted in cloning of the fbaA gene, which encodes a putative fructose 1,6-biphosphate aldolase (FBA), an enzyme involved in both glycolysis and gluconeogenesis. The fbaA1013 mutation is a chromosome rearrangement in the 5' regulatory region of the fbaA gene resulting in reduced or total loss of transcription in response to glycolytic and gluconeogenic carbon sources respectively. The fbaA gene is essential for growth. A functional FbaA protein is necessary for plasma membrane localization of the AgtA acidic amino acid (L-glutamate/L-aspartate) transporter, as the fbaA1013 mutation results in targeting to and presumably subsequent degradation of AgtA in the vacuole. Our results support a novel role of the FbaA protein that is, involvement in the regulation of amino acids transporters. PMID- 20026237 TI - Temperature independence of aquatic oxygen uptake in an air-breathing ectotherm and the implications for dive duration. AB - The thermal dependence of aerobic metabolic rate in air-breathing ectotherms indicates that an increase in temperature will reduce dive duration. The ability, however, to extract oxygen from the water provides an additional means to maintain aerobic metabolism and prolong submergence. Therefore, we hypothesised that as temperature increased, a bimodally respiring animal will compensate for the effects on aerobic metabolic rate by increasing aquatic oxygen uptake. The fully aquatic, bimodally respiring Arafura filesnake (Acrochordus arafurae) was used to determine how temperature affects the partitioning of oxygen exchange between aerial and aquatic sources and the impacts on dive duration. We found that rate of oxygen consumption increased with temperature (Q(10 (20-32 degrees C))=2.52) but aquatic oxygen uptake remained temperature independent and all extra oxygen demands were met by increasing aerial gas exchange, thus reducing dive duration. Maximum dive duration reduced from 77 min to 28 min between 20 degrees C and 32 degrees C. Under severe hypoxia, oxygen uptake from the water was negligible and dive duration was further reduced to 21 min at 32 degrees C. Despite dive duration being reduced as the water temperature increased, aquatic oxygen uptake was still responsible for significantly prolonging dive duration. PMID- 20026238 TI - Plasticity in food assimilation, retention time and coprophagy allow herbivorous cavies (Microcavia australis) to cope with low food quality in the Monte desert. AB - Energy balance depends on the efficiency with which organisms make use of their trophic resources, and has direct impact on their fitness. There are environmental variations that affect the availability as well as the quality of such resources; energy extraction also depends on the design of the digestive tract. It is expected that features associated with food utilization will be subjected to selective pressures and show some adjustment to the variability of the environment. Since energetic constraints challenge animals to display digestive compensatory mechanisms, the objective of this study is to determine the physiological and behavioral responses to spatial and seasonal heterogeneity in food quality. We investigated digestive strategies (digestive efficiency and coprophagy) in cavies inhabiting two different populations, and hence naturally experiencing different levels of diet quality. Cavies under experimentally different quality diets showed changes in dry matter digestibility and intake, digesta retention time and coprophagy. Our results partially support the expectations from theory and also reveal interpopulation differences in the ability to cope with changes in food quality, and may explain the capability of Microcavia australis to colonize extreme habitats. PMID- 20026239 TI - A guidance for renal biomarker lead optimization and use in translational pharmacodynamics. AB - Guidance for the use of biomarkers in pharmaceutical development and clinical trial optimization will reduce developmental cycle time. A 'fit-for-purpose' guidance for biomarker use is considered herein when the same biomarker is applied in very different contexts in drug development and after regulatory approval. Recent approved use of renal safety biomarkers in Good Laboratory Practice studies lacks sufficient guidance for the use of these markers across the drug development pipeline. In lead optimization, renal injury biomarkers are possible anchors for promising new prodromal metabolic biomarkers, which are applied before lead candidate selection. Renal injury biomarkers can now be evaluated as potential efficacy and pharmacodynamic biomarkers in clinical trial proof-of-concept studies for diabetic nephropathy. PMID- 20026241 TI - Metabolic control of puberty onset: new players, new mechanisms. AB - Puberty, as the end-point of a complex series of maturational events affecting the components of the hypothalamic-pituitary-gonadal (HPG) axis, is gated by the state of body energy reserves and sensitive to different metabolic cues; conditions of severe metabolic stress and energy unbalance (from anorexia to morbid obesity) being commonly linked to perturbation of the onset of puberty. In the last two decades, the neuroendocrine mechanisms responsible for the tight coupling between energy homeostasis and puberty onset have begun to be deciphered. These seemingly involve a plethora of metabolic hormones and neuropeptides, which impinge and integrate (mostly) at the hypothalamic centers governing reproduction. Yet, characterization of the mechanisms of action of such regulators (and even their nature and physiological relevance) still remains incomplete. In this review, we will summarize some recent developments in our knowledge of the effects and mechanisms of action of two key metabolic hormones, leptin and ghrelin, in the control of puberty onset. In addition, the roles of the hypothalamic Kiss1 system in the metabolic gating of puberty will be reviewed, with special attention to its regulation by leptin and the recent identification of the putative roles of Crtc1 and mTOR signaling as molecular conduits for the metabolic control of Kiss1 expression. Elucidation of these novel players and regulatory mechanisms will help for a better understanding of the determinants of the timing of puberty, and its eventual alterations in adverse metabolic conditions. PMID- 20026242 TI - Sarcocystis tupaia, sp. nov., a new parasite species employing treeshrews (Tupaiidae, Tupaia belangeri chinensis) as natural intermediate hosts. AB - The range of vertebrates that serve as intermediate hosts for parasites in the genus Sarcocystis remains incompletely defined. Here, we provide the first report of infections in treeshrews, describe the morphology of encysted parasites using light and transmission electron microscopy, and place this agent within a phylogenetic context by sequencing and comparing its 18S ribosomal DNA to that of related parasites. Muscle infections were diagnosed in four of 45 wild treeshrews captured in the vicinity of Kunming, Yunnan Province, Mainland China. Thread-like cysts (10.773+/-2.411mm in length, 0.106+/-0.009mm in width) had walls (0.538 0.746microm thick) that lacked perpendicular protrusions. The interior of the cyst was packed full with cyst merozoites, the shape of which was typical of Sarcocystis. The primary cyst wall consisted of a thin membrane supported by osmiophilic material, 31-60nm in thickness. The ground substance was about 105 526nm thickness. Cysts conformed to typical of 'type 1' sarcocysts. Freshly examined and frozen specimens did not differ in their cyst wall structure, however, the appearance of bradyzoites did differ: the conoid, rhoptries and micronemes were all visible in fresh bradyzoites; in stored bradyzoites, by contrast, the rhoptries appeared smaller, and although the conoid was visible, the micronemes were not. 18S rRNA gene was distinct from any previously reported sequence in GenBank. Their genetic and morphological uniformity suggest that these parasites, derived from treeshrews, represent a single biological species, Sarcocystis tupaia, sp. nov. PMID- 20026243 TI - Babesia sp. BQ1 (Lintan): molecular evidence of experimental transmission to sheep by Haemaphysalis qinghaiensis and Haemaphysalis longicornis. AB - Ovine babesiosis is an economically important disease induced by tick transmitted haemoparasites throughout the world. In China, several ovine Babesia strains have been isolated from field-collected ticks or sheep blood during the last two decades but little is known about the vector ticks and transmission pattern. Babesia sp. BQ1 (Lintan) is a Babesia strain infective for sheep and goats, isolated from blood of sheep experimentally infested with Haemaphysalis qinghaiensis collected in field. In the present study, we explored the experimental transmission of Babesia sp. BQ1 (Lintan) to sheep by H. qinghaiensis and Haemaphysalis longicornis. Based on the evidence from nested PCR, it suggested that H. qinghaiensis and H. longicornis are the potential vector ticks of Babesia sp. BQ1 (Lintan) and that larvae, nymphs and adults of both tick species were able to transmit Babesia sp. BQ1 (Lintan) to sheep. Parasites could be detected in the blood, by specific nested PCR, for one month post-infestation. PMID- 20026244 TI - Regulation of matrix metalloproteinase-9 and tissue plasminogen activator activity by alpha-synuclein in rat primary glial cells. AB - It is increasingly evident that neuroinflammatory response is involved in the pathogenesis of Parkinson's disease. In this study, we examined whether alpha synuclein, a major components of Lewy body that has been implicated in the modulation of neuroinflammation, regulates MMP-9 and tPA activity, which plays important roles in neurodegeneration as well as regeneration processes, in cultured rat primary glial cells. Monomeric alpha-synuclein dose-dependently increased MMP-9 but not MMP-2 activity as well as mRNA level from cultured rat primary astrocytes and microglial cells. Maximal stimulation was observed at 50 nM alpha-synuclein. In contrast, the activity of tPA was decreased by alpha synuclein with only marginal changes in the level of mRNA encoding tPA, if any. Interestingly, same concentration of alpha-synuclein aggregates did not induce MMP-9 activity. Overexpression of alpha-synuclein in rat primary astrocytes similarly increased MMP-9 activity. Treatment of alpha-synuclein increased the phosphorylation of ERK1/2 and the inhibition of ERK1/2 reversed the changes in MMP-9 and tPA activity. These results suggest further functional role of alpha synuclein via regulation of protease systems through modulation of ERK1/2 activity in brain. PMID- 20026245 TI - Trace amine metabolism in Parkinson's disease: low circulating levels of octopamine in early disease stages. AB - Recent evidence suggests that trace amines such as tyramine and octopamine, alternative products of tyrosine metabolism (an aminoacid parent of dopamine and noradrenaline), play a role in the homeostasis of the extrapyramidal system. However, the relevance of these trace amines in the pathogenesis of Parkinson's disease is still largely unknown. Here, we assessed the plasma levels of octopamine and noradrenaline in three sub-groups of PD patients, namely de novo, non-fluctuating and fluctuating patients, versus age-matched control subjects. We show that octopamine is detectable in plasma of all subjects, the mean levels of which are significantly lower in PD patients, including de novo patients, when compared to controls (p<0.001). Unlike this, no changes in plasmatic noradrenaline levels were found in the de novo patients, but only in plasma of fluctuating and non-fluctuating PD patients. These findings raise the possibility that Parkinson's disease is firstly characterized by abnormalities of tyrosine decarboxylase, rather than tyrosine hydroxylase, enzyme activity. Given the role of this enzyme in the production of trace amines, circulating octopamine levels may hold promise as a biomarker of early Parkinson's disease. PMID- 20026246 TI - The influence on cognitive processing from the switches of shooting angles in videos of real-world events: an ERP study. AB - This work mainly focuses on the influence from switches of shooting angles in videos during the cognitive processing in the human brain. In the experiment we used the videos with switches of shooting angles as materials and compared the ERPs elicited by the switch frames and the non-switch frames in the videos, it was found that when subjects were asked to pay attention to the video contents, the switch frames would trigger P3a-RON waveforms, but no N400 waveform was found in the ERP results. This showed that when subjects were concerned with the video contents, the switches of shooting angles would distract their attention from the video contents, but as long as the semantic meaning of the videos were coherent, the switches of shooting angles would not lead to significant difficulties in semantic comprehension. At the same time, the experimental results also further proved that the P3a and RON generally reflect the processing of task-irrelevant visual stimuli. PMID- 20026247 TI - Mild as well as severe insults produce necrotic, not apoptotic, cells: evidence from 60-min seizures. AB - We tested the hypothesis that mild insults produce apoptotic, and severe insults necrotic, cells by subjecting adult Wistar rats to 60-min instead of 3-h generalized seizures. Rats' brains were evaluated 6 and 24h later for evidence of neuronal necrosis by light and electron microscopy, the presence of TUNEL staining and active caspase-3 immunoreactivity, and for evidence of DNA laddering 24h after seizures. Apoptotic neurons from the retrosplenial cortex of postnatal day 8 rat pups served as positive controls. Six and 24h after seizures, 16 and 15 brain regions respectively out of 24 showed significant numbers of acidophilic neurons by hematoxylin and eosin stain. Three brain regions had significant numbers of TUNEL-positive neurons 24h after seizures. No neurons showed active caspase-3 immunoreactivity. Acidophilic neurons were necrotic by electron microscopic examination. Ultrastructurally, they were shrunken and electron dense, with shrunken, pyknotic nuclei and swollen mitochondria with disrupted cristae. Nuclei did not contain the irregular chromatin clumps found after 3-h seizures. None of the six brain regions studied ultrastructurally that show DNA laddering 24h after 3-h seizures showed DNA laddering 24h after 60-min seizures, probably because there were too few damaged neurons, although the lack of chromatin clumping might have been a contributing factor. Following seizures, a mild as well as a severe insult produces caspase-3-negative necrotic neurons. These results do not support the hypothesis that mild insults produce apoptotic, and severe insults, necrotic, cells. PMID- 20026248 TI - Simulated pathogenic conformational switch regions matched well with the biochemical findings. AB - Pathogenic conformational conversion is a general causation of many disease, such as transmissible spongiform encephalopathy (TSE) caused by misfolding of prion, sickle cell anemia, and etc. In such structural changes, misfolding occurs in regions important for the stability of native structure firstly. This destabilizes the normal conformation and leads to subsequent errors in folding pathway. Sites involved in the first stage can be deemed switch regions of the protein, and are vital for conformational conversion. Namely it could be a switch of disease at residue level. Here we report an algorithm that can identify such sites computationally with an accuracy of 93%, by calculating the probability of the native structure of a short segment jumping to a mistake one. Knowledge of such switch sites could be used to target clinical therapy, study physiological and pathologic mechanism of protein, and etc. PMID- 20026249 TI - Water transport between CNS compartments: functional and molecular interactions between aquaporins and ion channels. AB - The physiological ability of the mammalian CNS to integrate peripheral stimuli and to convey information to the body is tightly regulated by its capacity to preserve the ion composition and volume of the perineuronal milieu. It is well known that astroglial syncytium plays a crucial role in such process by controlling the homeostasis of ions and water through the selective transmembrane movement of inorganic and organic molecules and the equilibration of osmotic gradients. Astrocytes, in fact, by contacting neurons and cells lining the fluid filled compartments, are in a strategic position to fulfill this role. They are endowed with ion and water channel proteins that are localized in specific plasma membrane domains facing diverse liquid spaces. Recent data in rodents have demonstrated that the precise dynamics of the astroglia-mediated homeostatic regulation of the CNS is dependent on the interactions between water channels and ion channels, and their anchoring with proteins that allow the formation of macromolecular complexes in specific cellular domains. Interplay can occur with or without direct molecular interactions suggesting the existence of different regulatory mechanisms. The importance of molecular and functional interactions is pinpointed by the numerous observations that as consequence of pathological insults leading to the derangement of ion and volume homeostasis the cell surface expression and/or polarized localization of these proteins is perturbed. Here, we critically discuss the experimental evidence concerning: (1) molecular and functional interplay of aquaporin 4, the major aquaporin protein in astroglial cells, with potassium and gap-junctional channels that are involved in extracellular potassium buffering. (2) the interactions of aquaporin 4 with chloride and calcium channels regulating cell volume homeostasis. The relevance of the crosstalk between water channels and ion channels in the pathogenesis of astroglia-related acute and chronic diseases of the CNS is also briefly discussed. PMID- 20026250 TI - Development of a water-escape motivated version of the Stone T-maze for mice. AB - Mice provide a highly valuable resource for investigating learning and memory processes; however, many of the established tasks for evaluating learning and memory were developed for rats. Behaviors of mice in these tasks appear to be driven by different motivational factors, and as a result, they often do not perform reliably on tasks involving rewards traditionally used for rats. Because of difficulties in measuring learning and memory in mice as well as the need to have a task that can reliably measure these behavioral processes, we have developed a mouse version of the Stone T-maze utilizing what appears to be the primary motivation of mice, escape to a safe location. Specifically, we have constructed a task that requires the mouse to wade through water to reach a dark and dry goal box. To escape this aversive environment, the Stone T-maze requires learning the correct sequence of 13 left and right turns to reach the goal box. Through a series of experiments examining a variety of protocols, it was found that mice will reliably perform this task. This task can be used to assess learning and memory without the potential performance confounds that can affect performance of mice in other tasks. We believe this task offers a valuable new tool for evaluating learning and memory in mice not previously available to researchers. PMID- 20026251 TI - Pitx3-deficient aphakia mice display unique behavioral responses to psychostimulant and antipsychotic drugs. AB - The dorsal (A9) and ventral striatum (A10) of the midbrain mediate many of the effects of psychoactive drugs that alter emotion, cognition, and motor activity within the contexts of therapy or abuse. Although transgenic and knockout technologies have enabled development of genetic models to dissect contributions of specific dopamine (DA) receptor subtypes to psychoactive drug effects, few models exist that can distinguish contributions of A9 versus A10 circuits. Pitx3 is a transcription factor enriched in DA neurons. Aphakia (ak) mice deficient in Pitx3 show selective loss of nigrostriatal DA, while other DA pathways are relatively spared, and therefore could be a useful tool for investigating the role of this subclass of DA projections. We investigated the effects of stimulants amphetamine, apomorphine, and MK-801 and the antipsychotic drug haloperidol on behavior in ak mice. Whereas wild-type mice showed the characteristic locomotor hyperactivity in response to amphetamine (5 mg/kg) and apomorphine (4 mg/kg), these drugs caused a paradoxical suppression of locomotor hyperactivity in ak mice. MK-801 (0.2 mg/kg) induced hyperactivity was maintained in both wt and ak mice. Additionally, mutant but not wild-type mice were insensitive to the cataleptic effects of haloperidol (1 mg/kg). These studies indicate that the nigrostriatal DA circuit plays a critical role in maintaining normal responsiveness to psychotropic drugs that either stimulate or block DA neurotransmission. We propose that ak mice may represent a valuable genetic model not only to study Parkinson's disease, but also to dissect the pathophysiologic and pharmacotherapuetic mechanisms of other DA-mediated disorders such as attention-deficit hyperactivity disorder, drug abuse and schizophrenia. PMID- 20026252 TI - Site-specific action of L-3,4-dihydroxyphenylalanine in the striatum but not globus pallidus and substantia nigra pars reticulata evokes dyskinetic movements in chronic L-3,4-dihydroxyphenylalanine-treated 6-hydroxydopamine-lesioned rats. AB - Dyskinesia eventually develops in the majority of Parkinson's disease patients treated with l-3,4-dihydroxyphenylalanine (l-DOPA). We have investigated the effect of an acute and local administration of L-DOPA, GABA and glutamate to provoke dyskinetic movements in three basal ganglia structures (striatum, globus pallidus (GP) and substantia nigra pars reticulata (SNr)) of chronically L-DOPA treated, unilaterally 6-hydroxydopamine-lesioned rats. We demonstrated that L DOPA administration into the lesioned striatum using the technique of reverse in vivo microdialysis was an effective trigger to switch on dyskinesia. Notably, local L-DOPA perfusion at the same concentration in the ipsilateral GP and SNr did not provoke significant dyskinetic behaviour. Neither GABA nor glutamate triggered dyskinetic movements in the striatum, GP or SNr. We postulate a site specific action of L-DOPA for the evocation of already established dyskinesia since L-DOPA in the striatum but not in the GP or SNr switched on dyskinetic behaviour. PMID- 20026254 TI - Risk and reason in Barrett's esophagus. PMID- 20026253 TI - Acute noxious stimulation modifies morphine effect in serotonergic but not dopaminergic midbrain areas. AB - It is poorly understood if and how pain may modify the effect of opioids on neural systems that contribute to reward and addictive behavior. We hypothesized that the activation of ascending dopaminergic and serotonergic nuclei by morphine is modified by the presence of noxious stimulation. Immunohistochemical double labeling technique with Fos was used to examine if an intraplantar formalin injection, an acute noxious input, changed the effect of morphine on dopaminergic neurons of the ventral tegmental area (VTA), and serotonergic neurons of the dorsal raphe nucleus (DR). Four groups of rats were analyzed: (1) control injected with normal saline s.c., (2) rats treated with formalin into the hind paw 30 min after normal saline injection, (3) rats injected with morphine sulfate s.c., and (4) rats treated with formalin into the hind paw 30 min after morphine injection (morphine/formalin). Following morphine injection, there was an increase in the number of dopaminergic neurons in the VTA with Fos immunolabeling. However, noxious stimulation did not detectably change morphine's effect on Fos expression in VTA dopamine neurons. In contrast, the number of serotonergic neurons containing Fos was increased in the morphine/formalin group compared to all other groups and this effect was topographically selective for the dorsal area of the DR at mid rostro-caudal levels. Therefore, morphine's activation of the VTA, which is associated with motivated behavior and reward seeking, appears similar in the context of pain. However, activation of the ascending serotonin system, which influences mood and has the capacity to modify reward pathways, appears different. In addition, these findings reveal interactions between nociceptive signaling and opioids that contrasts with the notion that opioids simply block access of nociceptive signaling to supraspinal structures. PMID- 20026255 TI - Resveratrol is neuroprotective because it is not a direct activator of Sirt1-A hypothesis. AB - The plant polyphenol resveratrol (3,5,4'-trihydroxystilbene) has been touted to have multiple health benefits. A commonly cited mechanism of resveratrol action is via the activation of the longevity factor Sir2/Sirt1, whose deacetylase activity on several transcription factors has stress resistance and pro-survival effects. Resveratrol has been shown to be beneficial in various in vitro and in vivo models of central nervous system (CNS) neuron death and degeneration, presumably acting through Sirt1. However, accumulating recent evidence suggests that Sirt1 inhibitors are also neuroprotective. These contradictory results leave us with an apparently irreconcilable paradox. Based on other recent findings that resveratrol also activate AMP-activated protein kinase (AMPK), particularly in neurons, we hypothesize that reseveratrol does not exert its neuroprotective effect via direct Sirt1 activation. In fact, resveratrol is neuroprotective precisely because it does not activate Sirt1 during the acute phase of neuronal cell demise. However, its activation of AMPK may be neuroprotective. Furthermore, resveratrol may indirectly increase Sirt1 activity in recovering or spared cells via AMPK's elevation of NAD levels, which then translates into an overall beneficial outcome. The hypothesis could potentially be tested via selective AMPK silencing in various neuronal death and degeneration models, to see if the neuroprotective effect of resveratrol will be blunted. If proven true, the hypothesis has important ramifications in how reseveratrol, as well as novel Sirt1 activators, may be best used in treatment of CNS injuries and disorders. PMID- 20026256 TI - Polyoxyl 60 hydrogenated castor oil free nanosomal formulation of immunosuppressant Tacrolimus: pharmacokinetics, safety, and tolerability in rodents and humans. AB - OBJECTIVE: Develop Nanosomal formulation of Tacrolimus to provide safer alternative treatment for organ transplantation patients. Investigate safety, tolerability and pharmacokinetics of Nanosomal Tacrolimus formulation versus marketed Tacrolimus containing polyoxyl 60 hydrogenated castor oil (HCO-60) that causes side effects. METHODS: Nanosomal Tacrolimus was prepared in an aqueous system. The particle size was measured by Particle Sizing Systems and structure morphology was determined by freeze-fracture electron microscopy. Investigational safety studies were conducted in mice and rats. Safety and pharmacokinetics of Nanosomal Tacrolimus were also evaluated in healthy human subjects. RESULTS: The morphology of Nanosomal Tacrolimus showed a homogeneous population of nanosized particles with mean particle size of less than 100 nm. A 14 day consecutive administration of Nanosomal Tacrolimus up to 5 and 10mg/kg dose in rats and mice respectively, resulted in no mortality. Nanosomal Tacrolimus in human studies showed that it is safe and the pharmacokinetics profile is similar to the marketed HCO-60 based Tacrolimus. No significant change in peripheral blood lymphocyte percentage was noted in either mice or healthy human male subjects. CONCLUSIONS: Nanosomal Tacrolimus is well characterized product which provides a new treatment option. It contains no alcohol or surfactants like HCO-60. Thus, Nanosomal Tacrolimus presents a new and improved therapeutic approach for organ transplant patients compared to the marketed HCO-60 based Tacrolimus product. PMID- 20026257 TI - Characterisation of CpG methylation in the upstream control region of mouse Nat2: evidence for a gene-environment interaction in a polymorphic gene implicated in folate metabolism. AB - Human arylamine N-acetyltransferase 1 (NAT1), a polymorphic xenobiotic metabolising enzyme, has been investigated in relation to susceptibility and prognosis in certain types of cancer. Both human NAT1 and its murine equivalent NAT2 have previously been shown to play roles in the catabolism of folate, which is required for the synthesis of S-adenosylmethionine, the methyl donor for cellular methylation reactions. We have tested whether the expression of mouse Nat2 is subject to epigenetic regulation, specifically CpG methylation in the promoter region, by determining levels of 5-methylcytosine by bisulphite sequencing and methylation-specific PCR. Under normal conditions, methylation levels of the Nat2 promoter were low, and varied in different tissues. However, CpG methylation was significantly increased by dietary folate supplementation, and increased methylation corresponded to decreased use of the core promoter. Functional deletion of the Nat2 gene gave rise to a significant increase in Nat2 methylation, extending our previous observations that folate catabolism is decreased in Nat2 null mice. Mouse NAT2 is likely to influence epigenetic gene control, particularly of its own locus, and this is consistent with recent evidence associating aberrant mouse Nat2/human NAT1 gene expression with certain developmental malformations and cancers. PMID- 20026258 TI - Geoepidemiology: the environment and spondyloarthropathies. AB - Spondyloarthropathies (SPA) are a group of common inflammatory rheumatic disorders characterized by axial and or peripheral arthritis, associated with enthesitis, dactylitis and potential extra-articular manifestations such as uveitis and skin rash. The diseases which comprise the group, share a common genetic predisposition, the HLA-B27 gene, however this association varies markedly among the various SPAs and among different ethnic groups. Environmental factors seem to be triggering the diseases in the genetically predisposed. The radiographic hallmark of the group is sacroiliitis, which when present is of help in the diagnosis. Various sets of diagnostic and classification criteria were developed over the years with the latest European Spondyloarthropathy Study Group (ESSG) criteria which are the most widely used. MRI changes have been included in the new classification criteria of early axial SPA and are now considered as a major tool in the diagnosis. Until recent years, there were no real disease modifying anti-rheumatic drugs which were able to halt the disease progression. Tumor necrosis factor (TNF)-alfa blocking agents, have now become the mainstream of therapy providing the patients an effective treatment option. PMID- 20026260 TI - Dipeptidyl peptidase 9 (DPP9) from bovine testes: identification and characterization as the short form by mass spectrometry. AB - The dipeptidyl peptidases (DPP) 8 and 9 belong to the DPP4 activity and/or structure homologues (DASH). Recently, a DPP9-like protein was purified from bovine testes. The aim of the present study was to prove its identity and to investigate the characteristics of this natural enzyme. We report the identification and N-terminal sequence analysis by MALDI-TOF/TOF MS, of the purified bovine enzyme as DPP9. The tryptic peptides after in-gel digestion covered 41% and 38% of the short and full-length variants of bovine DPP9, respectively. Using Asp-N digestion combined with a very recently described mass spectrometric method using DITC glass beads, the N-terminal peptide (XTGALTSERG) was isolated. It corresponds to the N-terminus of the short form of bovine DPP9. There was no evidence for glycosylation of purified bovine DPP9. The purified DPP9 was activated and stabilized by DTT. Bovine DPP9 lost its activity almost completely after alkylation with N-ethylmaleimide. Also alkylation with iodoacetamide inhibited DPP9, albeit only 70%. Other properties of bovine DPP9 are reported, including functional stability and sensitivity towards metal ions. Our results indicate that the short form of DPP9 can be isolated from bovine testes and that it behaves as a stable enzyme suitable for further functional and biochemical characterization as well as for inhibitor screening and characterization. PMID- 20026259 TI - The role of NOX enzymes in ethanol-induced oxidative stress and apoptosis in mouse embryos. AB - Reactive oxygen species (ROS) play an important role in ethanol-induced apoptosis and teratogenesis. However, the major sources of ROS in ethanol-exposed embryos have remained undefined. This study was conducted to determine the role of NADPH oxidase (NOX) in ethanol-induced oxidative stress and apoptosis in mouse embryos. Analyses of mRNA expression indicated that ethanol treatment resulted in a significant increase in mRNA expression of NOX catalytic subunit Duox-1 in gestational day 9 (GD 9:0) mouse embryos. Ethanol exposure also resulted in significant increases in mRNA expression of NOX regulatory subunits, p22phox, p67phox, NOXA1 and NOXO1. In addition, a significant increase in NOX enzyme activity was found in the ethanol-exposed embryos as compared to controls. Co treatment with the NOX inhibitor, diphenyleneiodonium (DPI), significantly prevented ethanol-induced increases in NOX enzyme activity, ROS generation and oxidative DNA damage in ethanol-exposed embryos. DPI treatment also resulted in a reduction in caspase-3 activation, decreased caspase-3 activity and diminished prevalence of apoptosis in ethanol-exposed embryos. These results support the hypothesis that NOX is a critical source of ROS in ethanol-exposed embryos and that it plays an important role in ethanol-induced oxidative stress and pathogenesis. PMID- 20026261 TI - Aerodynamic deposition of combination dry powder inhaler formulations in vitro: a comparison of three impactors. AB - Inertial impaction is generally regarded as the 'gold standard' for the in vitro assessment of aerodynamic deposition of inhaled formulations. Despite the availability of several impactors, few studies have compared measurements of aerodynamic deposition using multiple impactors and none employed a combination formulation. The aerodynamic deposition of the combination dry powder inhaler (DPI) Seretide Accuhaler, which contains salmeterol xinafoate (SX) and fluticasone propionate (FP), was assessed using the Andersen cascade impactor (ACI), multi-stage liquid impinger (MSLI) and next generation impactor (NGI) and the results were compared. Two Seretide products were tested at flow rates of 30 and QLmin(-1), the latter corresponding to a pressure drop of 4kPa across the device. Significant differences in the particle size distributions were observed when the same formulation was tested using various impactors. The ACI was found to be less suitable for DPI testing at flow rates considerably higher than 28.3Lmin(-1) due to the significant overlap in the cut-off curves of the pre separator and stage 0. This was not the case with the MSLI but the data derived were limited by the relatively small number of stages. Deposition data determined by the three impactors were significantly different. The NGI produced good resolution and minimal inter-stage overlap and was regarded as the impactor of choice for DPI testing. PMID- 20026262 TI - Exploring the venom proteome of the African puff adder, Bitis arietans, using a combinatorial peptide ligand library approach at different pHs. AB - We report the 2DE-based proteomic characterization of the venom of the medically important African puff adder, Bitis arietans, after prefractionation by incubation with a solid-phase combinatorial hexapeptide ligand library (CPLL) at three different pH values. This approach yielded partially overlapping yet clearly distinct sets of proteins. The B. arietans venom proteome, merged from the four sets of proteins comprises at least 43 distinct proteins from 9 toxin families. In line with a previous reverse-phase HPLC-based venomic characterization on the same species, SVMPs, serine proteinases, C-type lectin like proteins, and to a minor extent PLA(2), disintegrin bitistatin, and cystatin, comprise the major toxins in the venom of B. arietans. However, the 2D CPLL approach employed here identified both a significantly higher (about double) number of proteins than a previous venomic approach, and many very minor components barely, or not at all, detectable in the 2DE separation of whole venom. 30 proteins from the CPLL-merged venom proteome matched some of the 63 toxin clusters generated by sequencing one thousand randomly selected venom gland cDNA library clones of the same species. The low (47%) concordance between transcriptome and proteome may be interpreted in terms of intraspecific venom variation. Comparison of the reverse-phase HPLC separations of the venom proteins of B. arietans from Ghana and Nigeria supports this view. PMID- 20026263 TI - GDP-capture compound--a novel tool for the profiling of GTPases in pro- and eukaryotes by capture compound mass spectrometry (CCMS). AB - The functional isolation of proteome subsets based on small molecule-protein interactions is an increasingly popular and promising field in functional proteomics. Entire protein families may be profiled on the basis of their common interaction with a metabolite or small molecule inhibitor. This is enabled by novel multifunctional small molecule probes. One platform approach in this field are Capture Compounds that contain a small molecule of interest to bind target proteins, a photo-activatable reactivity function to covalently trap bound proteins, and a sorting function to isolate Capture Compound-protein conjugates from complex biological samples for direct trypsinisation and protein identification by liquid chromatography/mass spectrometry (CCMS). We here present the synthesis and application of a novel GDP-Capture Compound for the functional enrichment of GTPases, a pivotal protein family that exerts key functions in signal transduction. We present data from CCMS experiments on two biological lysates from Escherichia coli and from human-derived Hek293 cells. The GDP Capture Compound robustly captures a wide range of different GTPases from both systems and will be a valuable tool for the proteomic profiling of this important protein family. PMID- 20026264 TI - Postnatally induced inactivation of Osterix in osteoblasts results in the reduction of bone formation and maintenance. AB - Osterix (Osx) is a zinc-finger-containing transcription factor that is highly specific to osteoblasts in vivo. Because Osx homozygous null mutants die in the immediate perinatal period showing a complete absence of bone formation, it is impossible determine the role that Osx plays in bones that have already formed after birth. To determine whether Osx is essential for bone maintenance and homeostasis, we conditionally inactivated the Osx gene in adult bone using the Cre/loxP recombination system. In previous reports, 2.3-kb Col1a1-CreERT2 mice that expressed a Cre recombinase that is transiently inducible by 4 hydroxytamoxifen (4-OHT) were intercrossed with Rosa26R (R26R) reporter mice, which resulted in the production of Cre-expressing osteoblasts that were detected upon X-gal staining. In the present study, inducible Col1a1-CreERT2 transgenic mice and conditional Osx mice (Osx(flox/+)) were used to generate Osx(flox/ );Col1a1-CreERT2 mice. The Osx gene in Osx(flox/-);Col1a1-CreERT2 mice was inactivated in the osteoblasts of already formed bones by active Cre recombinase after the administration of 4-OHT. The bones from 4-OHT-treated Osx(flox/ );Col1a1-CreERT2 mice and oil-treated control mice were analyzed by radiography, histology, and histomorphometry. Even though no significant difference was observed in the radiographic images of the whole mouse skeletons, the mineralized trabecular bone volume and number in lumbar vertebrae were remarkably reduced in 4-OHT-treated Osx(flox/-);Col1a1-CreERT2 mice. In addition, the rate of bone formation and area of mineralized surface were also reduced in 4-OHT-treated Osx(flox/-);Col1a1-CreERT2 mice. Osx inactivation in already formed bones during the postnatal period caused a functional defect in osteoblasts that was followed by a reduction of bone formation, even though there were no apparent differences in osteoblast proliferation and osteoclast formation. Taken together, these results indicate that Osx is required to maintain osteoblast function following adult bone maintenance. PMID- 20026265 TI - Five years treatment with strontium ranelate reduces vertebral and nonvertebral fractures and increases the number and quality of remaining life-years in women over 80 years of age. AB - INTRODUCTION: Longevity has resulted in a greater proportion of the population entering a time of life when increasing bone fragility and falls predispose to fractures, particularly nonvertebral fractures. Women over 80 years of age constitute 10% of the population but contribute 30% of all fractures and 60% of all nonvertebral fractures. Despite this, few studies have examined antifracture efficacy of treatments in this high-risk group and none has provided evidence for benefits beyond 3 years. MATERIALS AND METHODS: To determine whether strontium ranelate reduces the risk of vertebral and nonvertebral fractures during 5 years, we analyzed a subgroup of 1489 female patients over 80 years of age (mean 83.5+/ 3.0 years) with osteoporosis from the SOTI (spinal osteoporosis therapeutic intervention) and TROPOS (treatment of peripheral osteoporosis) studies randomized to strontium ranelate 2 g/d or placebo. All received a supplement of calcium plus vitamin D. RESULTS: By intention to treat, vertebral fracture risk was reduced by 31% (relative risk, RR=0.69; 95% confidence interval, CI 0.52 0.92), nonvertebral fracture risk by 27% (RR=0.73; 95% CI 0.57-0.95), major nonvertebral fracture risk by 33% (RR=0.67; 95% CI 0.50-0.89) and hip fracture risk by 24% (RR=0.76; 95% CI 0.50-1.15, not significant). Treatment was cost saving as it decreased cost and increased QALYs and life-years. DISCUSSION: Strontium ranelate safely produced a significant reduction in vertebral and nonvertebral fracture risk during 5 years in postmenopausal women over 80 years of age and was cost saving. PMID- 20026266 TI - Nonlinearity in eardrum vibration as a function of frequency and sound pressure. AB - It is generally accepted that the middle ear acts mainly as a linear system for sound pressures up to 130 dB SPL in the auditory frequency range. However, at quasi-static pressure loads a strong nonlinear response has been demonstrated. Consequently, small nonlinear distortions may also be present in the middle ear response in the auditory frequency range. A new measurement method was developed to quickly determine vibration response, nonlinear distortions and noise level of acoustically driven biomechanical systems. Specially designed multisines are used for the excitation of the test system. The method is applied on a gerbil eardrum for sound pressures ranging from 90 to 120 dB SPL and for frequencies ranging from 125 Hz to 16 kHz. The experiments show that nonlinear distortions rise above noise level at a sound pressure of 96 dB SPL, and they grow as sound pressure increases. Post-mortem changes in the middle ear influence the nonlinear distortions rapidly until a stabilization occurs after approximately 3h. PMID- 20026267 TI - Heterogeneous distribution of SNPs in the human genome: microsatellites as predictors of nucleotide diversity and divergence. AB - Understanding the forces that govern the distribution of single nucleotide polymorphisms is vital for many of their applications. Here we conducted a systematic search to quantify how both SNP density and human-chimpanzee divergence vary around different repetitive sequences. We uncovered a highly complicated picture in which these quantities often differ significantly from the genome-wide average in regions extending more than 20 kb, the direction of the deviation varying with repeat number and motif. AT microsatellites in particular are potent predictors of SNP density, long (AT)(n) repeat tracts tending to be found in regions of significantly reduced SNP density and low GC content. Although the causal relationships remain difficult to determine, our results indicate a strong relationship between microsatellites and the DNA that flanks them. Our results help to explain the mixed picture that emerges from other studies and have important implications for the way in which genetic diversity is distributed in our genomes. PMID- 20026268 TI - A negative regulatory role for Y1111 on the Tie-2 RTK. AB - Tie2 is a receptor tyrosine kinase (RTK) essential for aspects of both normal and pathological angiogenesis. Understanding how this receptor is regulated is important for development of therapeutic angiogenic agents. Evidence suggests the C-terminal tail of the receptor plays a negative regulatory role in Tie2 signaling and function. Here we investigated the role of a specific C-tail residue, Y1111, in Tie2 signaling by generating a number of receptor point mutants. We found that mutation of this site to phenylalanine (Y1111F) results in an increase in receptor phosphorylation and kinase activity, as well as increased downstream signaling. Furthermore, mutation of Y1111 to the highly charged aspartate (Y1111D) or glutamate (Y1111E) results in even more dramatic increase in receptor phosphorylation and activity. Limited protease digestion studies indicate that these mutations may alter receptor conformation and potentially relieve negative inhibition imparted by the C-tail of Tie2. These studies suggest that Y1111 plays a key role in negative regulation of Tie2 activity and they provide important insight into molecular mechanisms behind the intrinsic ability of this RTK to regulate its own activity. PMID- 20026269 TI - Calpains and proteasomes mediate degradation of ryanodine receptors in a model of cardiac ischemic reperfusion. AB - Type-2 ryanodine receptors (RyR2)--the calcium release channels of cardiac sarcoplasmic reticulum--have a central role in cardiac excitation-contraction coupling. In the heart, ischemia/reperfusion causes a rapid and significant decrease in RyR2 content but the mechanisms responsible for this effect are not fully understood. We have studied the involvement of three proteolytic systems- calpains, the proteasome and autophagy--on the degradation of RyR2 in rat neonatal cardiomyocyte cultures subjected to simulated ischemia/reperfusion (sI/R). We found that 8h of ischemia followed by 16h of reperfusion decreased RyR2 content by 50% without any changes in RyR2 mRNA. Specific inhibitors of calpains and the proteasome prevented the decrease of RyR2 caused by sI/R, implicating both pathways in its degradation. Proteasome inhibitors also prevented the degradation of calpastatin, the endogenous calpain inhibitor, hindering the activation of calpain induced by calpastatin degradation. Autophagy was activated during sI/R as evidenced by the increase in LC3-II and beclin-1, two proteins involved in autophagosome generation, and in the emergence of GFP LC3 containing vacuoles in adenovirus GFP-LC3 transduced cardiomyocytes. Selective autophagy inhibition, however, induced even further RyR2 degradation, making unlikely the participation of autophagy in sI/R-induced RyR2 degradation. Our results suggest that calpain activation as a result of proteasome-induced degradation of calpastatin initiates RyR2 proteolysis, which is followed by proteasome-dependent degradation of the resulting RyR2 fragments. The decrease in RyR2 content during ischemia/reperfusion may be relevant to the decrease of heart contractility after ischemia. PMID- 20026270 TI - Transformation of some 3alpha-substituted steroids by Aspergillus tamarii KITA reveals stereochemical restriction of steroid binding orientation in the minor hydroxylation pathway. AB - Aspergillus tamarii contains an endogenous lactonization pathway which can transform progesterone to testololactone in high yield through a sequential four step enzymatic pathway. In this pathway testosterone is formed which primarily undergoes oxidation of the C-17beta-alcohol to a C-17 ketone but, can also enter a minor hydroxylation pathway where 11beta-hydroxytestosterone is produced. It was recently demonstrated that this hydroxylase could monohydroxylate 3beta hydroxy substituted saturated steroidal lactones in all four possible binding orientations (normal, reverse, inverted normal, inverted reverse) on rings B and C of the steroid nucleus. It was therefore of interest to determine the fate of a series of 3alpha-substituted steroidal analogues to determine stereochemical effect on transformation. Hydroxylation on the central rings was found to be restricted to the 11beta-position (normal binding), indicating that the 3alpha stereochemistry removes freedom of binding orientation within the hydroxylase. The only other hydroxylation observed was at the 1beta-position. Interestingly the presence of this functional group did not prevent lactonization of the C-17 ketone. In contrast the presence of the 11beta-hydroxyl completely inhibited Baeyer-Villiger oxidation, a result which again demonstrates that single functional groups can exert significant control over metabolic handling of steroids in this organism. This may also explain why lactonization of 11beta hydroxytestosterone does not occur. Lactonization of the C-17 ketone was not significantly affected by the 3alpha-alcohol with significant yields achieved (53%). Interestingly a time course experiment demonstrated that the presence of the 3alpha-acetate inhibited the Baeyer-Villiger monooxygenase with its activity being observed 24h later than non-acetate containing analogues. Apart from oxidative transformations observed a minor reductive pathway was revealed with the C-17 ketone being reduced to a C-17beta-alcohol for the first time in this organism. PMID- 20026272 TI - Urokinase-type plasminogen activator regulates neurodegeneration and neurogenesis but not vascular changes in the mouse hippocampus after status epilepticus. AB - Expression of urokinase-type plasminogen activator (uPA) is increased after brain injury, suggesting that, like in cancer tissue, uPA plays roles in brain remodeling. Here we injured brain with intrahippocampal kainic acid (KA) injection in adult Wt and uPA-/- mice. At 20 days post-injury, uPA-/- mice had more severe loss of contralateral pyramidal (p<0.05) and hilar neurons (p<0.05) than Wt mice. The number of doublecortin (DCX)-positive newly born neurons was also reduced in uPA-/- mice as compared to Wt (p<0.01). No difference was observed in granule cell dispersion or distribution of DCX-positive neurons in the dentate gyrus. uPA deficiency did not affect the total length of hippocampal blood vessels or vessel density. No differences were observed in the severity of status epilepticus or consequent epilepsy between the genotypes. These data indicate that uPA deficiency can unfavorably modulate both delayed neurodegeneration and neurogenesis but has little effect on post-injury neuronal migration and vascular density. Our results favor the idea that elevated uPA during the post-injury phase is neuroprotective. PMID- 20026271 TI - Ethyl pyruvate protects against hypoxic-ischemic brain injury via anti-cell death and anti-inflammatory mechanisms. AB - Ethyl pyruvate (EP) is protective in experimental models of many illnesses. This study investigates whether EP can protect against neonatal hypoxic-ischemic (H-I) brain injury. Pre-treatment with EP significantly reduced brain damage at 7 days post-H-I, with 50 mg/kg EP achieving over 50% recovery in tissue loss compared to vehicle-treated animals. Delayed treatment with EP until 30 min after H-I was still neuroprotective. EP-afforded brain protection, together with neurological function improvement, was observed up to 2 months after H-I. We further demonstrated an inhibitory effect of EP on cell death, both in an in vivo model of H-I and in in vitro neuronal cultures subjected to OGD, by reducing calpain activation and calcium dysregulation. Moreover, EP exerted an anti-inflammatory effect in microglia by inhibiting NF-kappaB activation and subsequent release of inflammatory mediators. Taken together, our results suggest that EP confers potent neuroprotection against neonatal H-I brain injury via its anti-cell death and anti-inflammatory actions. EP is a potential novel therapeutic agent for neonatal H-I brain injury. PMID- 20026273 TI - The cyclin-dependent kinase inhibitor PHA-848125 suppresses the in vitro growth of human melanomas sensitive or resistant to temozolomide, and shows synergistic effects in combination with this triazene compound. AB - PHA-848125 is a novel cyclin-dependent kinase inhibitor under Phase I/II clinical investigation. In this study, we describe, for the first time, the effect of PHA 848125 on human melanoma cells in vitro. Seven melanoma cell lines with different sensitivity to temozolomide (TMZ) were exposed to PHA-848125 for 5 days and then assayed for cell growth. In all cases, including TMZ-resistant cells, PHA-848125 IC(50) values were significantly below the maximum plasma concentrations achievable in the clinic. In the most PHA-848125-sensitive cell line, the drug caused a concentration-dependent G(1) arrest. PHA-848125 also impaired phosphorylation of the retinoblastoma protein at CDK2 and CDK4 specific sites, decreased retinoblastoma protein and cyclin A levels, and increased p21(Cip1), p27(Kip1) and p53 expression. Combined treatment with fixed ratios of TMZ plus PHA-848125 was studied in three melanoma cell lines. PHA-848125 was added to the cells 48 h after TMZ and cell growth was evaluated after 3 additional days of culture. Parallel experiments were performed in the presence of O(6) benzylguanine (BG), to prevent repair of methyl adducts at O(6)-guanine induced by TMZ. Drug combination of TMZ plus BG and PHA-848125 produced additive or synergistic effects on cell growth, depending on the cell line. In the absence of BG, the combination was still more active than the single agents in the cell line moderately sensitive to TMZ, but comparable to PHA-848125 alone in the two TMZ resistant cell lines. When TMZ plus BG were used in combination with PHA-848125 against cultured normal melanocytes, neither synergistic nor additive antiproliferative effects were observed. Our results indicate that PHA-848125 can have a therapeutic potential in melanoma patients, alone or combined with TMZ. Moreover this agent appears to be particularly attractive on the bases of its effectiveness against TMZ-resistant melanoma cells. PMID- 20026275 TI - Effect of curcumin on brain insulin receptors and memory functions in STZ (ICV) induced dementia model of rat. AB - Curcumin, the principal curcuminoid of turmeric, exhibits beneficial role in several neurodegenerative disorders such as dementia of Alzheimer type. Recent evidences suggest the involvement of brain insulin receptors (IRs) in the pathophysiology of dementia disorders. Therefore, the present study was undertaken to investigate the effect of curcumin on memory functions, brain IRs, acetylcholinesterase (AChE) activity and oxidative stress in intracerebroventricular (ICV) administered streptozotocin (STZ) induced dementia in rats. Rats were injected with STZ (3 mg/kg, ICV) bilaterally twice, on day 1 and 3 and curcumin (200 mg/kg, po) was administered in pre- and post-treatment schedules. STZ (ICV) treated group had shown memory deficit as indicated by no significant decrease in latency time in Morris water maze test and significant decrease in IR protein level in both hippocampus and cerebral cortex. Pre- and post-treatment of curcumin in STZ (ICV) treated rats significantly restored the memory deficit and IR protein level in both the regions. Furthermore, STZ (ICV) resulted into enhanced AChE activity in hippocampus and cerebral cortex which was normalized by curcumin pre- and post-treatment. An increase in MDA level and decrease in GSH level were obtained in both hippocampus and cerebral cortex in STZ treated group, indicating state of oxidative stress, which was also attenuated by pre- and post-treatment of curcumin. The results suggest that besides the anticholinesterase and antioxidant activity, effect on brain IR may also be an important factor for protective effect of curcumin against STZ induced dementia model. PMID- 20026274 TI - Protein deacetylation by SIRT1: an emerging key post-translational modification in metabolic regulation. AB - The biological function of most proteins relies on reversible post-translational modifications, among which phosphorylation is most prominently studied and well recognized. Recently, a growing amount of evidence indicates that acetylation deacetylation reactions, when applied to crucial mediators, can also robustly affect the function of target proteins and thereby have wide-ranging physiological impacts. Sirtuin 1 (SIRT1), which functions as a nicotinamide adenine dinucleotide (NAD(+))-dependent protein deacetylase, deacetylates a wide variety of metabolic molecules in response to the cellular energy and redox status and as such causes significant changes in metabolic homeostasis. This review surveys the evidence for the emerging role of SIRT1-mediated deacetylation in the control of metabolic homeostasis. PMID- 20026276 TI - Neuropotent self-renewing neural stem (NS) cells derived from mouse induced pluripotent stem (iPS) cells. AB - Neural stem (NS) cells are a homogenous population of stem cells that expands in monolayer under serum-free conditions while remaining highly neuropotent. Here, we generated NS cells from induced pluripotent stem (iPS) cells that were previously derived from mouse fibroblasts (NS-(f)iPS). We showed that NS-(f)iPS cells exhibit long-term expansion and express markers of neurogenic radial glia. Analyses of the regional markers expressed in NS-(f)iPS cells suggested a ventral rhombencephalic identity. Upon exposure to differentiation protocols, NS-(f)iPS cells produce neurons, astrocytes, and oligodendrocytes with an efficiency similar to ES-derived NS cells. NS-(f)iPS cells represent a new tool for studying neural cell fate determination and terminal differentiation, providing an interesting resource for experimental transplantation. Comparative studies between NS cells derived from iPS cells, reprogrammed from different somatic sources, and from authentic ES cells are necessary to identify critical elements for multipotency acquisition. PMID- 20026277 TI - Expression, purification and direct eletrochemistry of cytochrome P450 6A1 from the house fly, Musca domestica. AB - A plasmid (pCW) was modified to code for the complete sequence of house fly (Musca domestica) cytochrome P450 6A1 (CYP6A1) with only the second amino acid changed in the N-terminal portion and this plasmid was used to express the enzyme CYP6A1 in Escherichia coli cells. With the addition of delta-aminolevulinic acid and FeCl(3) to the culture, the enzyme was produced at a level about 0.25 micromol L(-1) (15mgL(-1)) of culture with approximately 50% of the P450 being associated with the membrane fraction. The CYP6A1 protein was characterized and the content of CYP6A1 in each fraction was determined by the spectroscopic method. A nearly homogenous CYP6A1 was obtained by purification with a combination of DEAE Sepharose fast flow and hydroxyapatite chromatography. Direct electrochemistry of CYP6A1 in a didodecyldimethylammonium bromide (DSAB) film on an edge-plane pyrolytic graphite electrode (EPG) has been obtained and the catalytic activity of the enzyme to aldrin has been demonstrated by the cyclic voltammetry. PMID- 20026278 TI - Motor imagery in response to fake feedback measured by functional near-infrared spectroscopy. AB - The objective of this study was to describe brain oxygenation patterns during motor imagery (MI) in response to feedback using functional near-infrared spectroscopy (fNIRS). fNIRS was recorded over the primary motor cortex in 15 healthy subjects using a right hand motor task during four fake feedback conditions: MI without feedback (MI(0)), MI with positive (MI(+)) and negative feedback (MI(-)) and during actual movement execution (ME) as control task. Behavioral data were collected using the Edinburgh Handedness Inventory (EHI) and The Kinesthetic and Visual Imagery Questionnaire (KVIQ-10). We observed inter condition differences and inter-subject variability in signal amplitude with larger O(2)Hb concentration changes both in response to MI(+) (0.154+/-0.067 microM) and MI(-) (0.129+/-0.074 microM) as compared to MI(0) (0.109+/-0.024 microM) and ME (0.210+/-0.013 microM). We present fNIRS data of MI performance in response to different feedback conditions indicating that there exist distinct oxygenation patterns. These data may contribute to the development of fNIRS controlled feedback systems. PMID- 20026279 TI - The effect of filtering on Granger causality based multivariate causality measures. AB - In the past, causality measures based on Granger causality have been suggested for assessing directionality in neural signals. In frequency domain analyses (power or coherence) of neural data, it is common to preprocess the time series by filtering or decimating. However, in other fields, it has been shown theoretically that filtering in combination with Granger causality may lead to spurious or missed causalities. We investigated whether this result translates to multivariate causality methods derived from Granger causality with (a) a simulation study and (b) an application to magnetoencephalographic data. To this end, we performed extensive simulations of the effect of applying different filtering techniques and evaluated the performance of five different multivariate causality measures in combination with two numerical significance measures (random permutation and leave one out method). The analysis included three of the most widely used filters (high-pass, low-pass, notch filter), four different filter types (Butterworth, Chebyshev I and II, elliptic filter), variation of filter order, decimating and interpolation. The simulation results suggest that preprocessing without a strong prior about the artifact to be removed disturbs the information content and time ordering of the data and leads to spurious and missed causalities. Only if apparent artifacts like a current or movement artifact are present, filtering out the respective disturbance seems advisable. While oversampling poses no problem, decimation by a factor greater than the minimum time shift between the time series may lead to wrong inferences. In general, the multivariate causality measures are very sensitive to data preprocessing. PMID- 20026280 TI - Investigating the effect of blood susceptibility on phase contrast in the human brain. AB - Recent work has shown a dramatic contrast between GM and WM in gradient echo phase images at high field (7 T). Although this contrast is key to the exploitation of phase in imaging normal and pathological tissue, its origin remains contentious. Several sources for this contrast have been considered including iron content, myelin, deoxy-hemoglobin, or water-macromolecule interactions. Here we quantify the contribution of intravascular dHb to the GM/WM contrast in the human brain at 7 T by modulating the susceptibility of the blood using a paramagnetic contrast agent. By carrying out high resolution, dynamic, gradient echo imaging before, during and after the injection of the contrast agent, we were able to follow the change in GM/WM phase contrast and to monitor simultaneously the susceptibility of the blood. Using these data in conjunction with the known susceptibility of venous blood we estimate the upper bound for the relative contribution of dHb in the vasculature to the measured GM/WM phase contrast to be 0.48 Hz for GM close to the pial surface, and 0.27 Hz for deeper GM. These values are up to 20% of the GM/WM phase difference observed in the human brain at 7 T. Furthermore, we found that the fractional blood volume differences required to account for the observed GM/WM phase contrast are 1.3% and 0.7% for GM close to the pial surface and for deeper GM, respectively. PMID- 20026281 TI - Model-driven parameterization of the cortical surface for localization and inter subject matching. AB - In this paper we present a generic and organized model of cortical folding, and a way to implement this model on any given cortical surface. This results in a model-driven parameterization, providing an anatomically meaningful coordinate system for cortical localization, and implicitly defining inter-subject surface matching without any deformation of surfaces. We present our cortical folding model and show how it naturally defines a parameterization of the cortex. The mapping of the model to any given cortical surface is detailed, leading to an anatomically invariant coordinate system. The process is evaluated on real data in terms of both anatomical and functional localization, and shows improved performance compared to a traditional volume-based normalization. It is fully automatic and available with the BrainVISA software platform. PMID- 20026283 TI - LipL53, a temperature regulated protein from Leptospira interrogans that binds to extracellular matrix molecules. AB - The regulation of gene expression by environmental signals, such as temperature and osmolarity, has been correlated with virulence. In this study, we characterize the protein LipL53 from Leptospira interrogans, previously shown to react with serum sample of individual diagnosed with leptospirosis and to be up regulated by shift to physiological osmolarity. The recombinant protein was expressed in Escherichia coli system, in insoluble form, recovered by urea solubilization and further refolded by decreasing the denaturing agent concentration during the purification procedure. The secondary structure content of the recombinant LipL53, as assessed by circular dichroism, showed a mixture of beta-strands and alpha-helix. The presence of LipL53 transcript at 28 degrees C was only detected within the virulent strains. However, upon shifted of attenuated cultures of pathogenic strains from 28 degrees C to 37 degrees C and to 39 degrees C, this transcript could also be observed. LipL53 binds laminin, collagen IV, cellular and plasma fibronectin in dose-dependent and saturable manner. Animal challenge studies showed that LipL53, although immunogenic, elicited only partial protection in hamsters. LipL53 is probably surface exposed as seen through immunofluorescence confocal microscopy. Our results suggest that LipL53 is a novel temperature regulated adhesin of L. interrogans that may be relevant in the leptospiral pathogenesis. PMID- 20026282 TI - Life and death of a BiP substrate. AB - BiP is the mammalian endoplasmic reticulum (ER) Hsp70 orthologue that plays a major role in all functions of this organelle including the seemingly opposing functions of aiding the maturation of unfolded nascent proteins and identifying and targeting chronically unfolded proteins for degradation. The recent identification of mammalian BiP co-factors combined with delineation of the ER degradation machinery and data suggesting that the ER is subdivided into unique regions helps explain how these different functions can occur in the same organelle and raises some unresolved issues. PMID- 20026284 TI - Differential expression of choline kinase isoforms in skeletal muscle explains the phenotypic variability in the rostrocaudal muscular dystrophy mouse. AB - Choline kinase in mammals is encoded by two genes, Chka and Chkb. Disruption of murine Chka leads to embryonic lethality, whereas a spontaneous genomic deletion in murine Chkb results in neonatal forelimb bone deformity and hindlimb muscular dystrophy. Surprisingly, muscular dystrophy isn't significantly developed in the forelimb. We have investigated the mechanism by which a lack of choline kinase beta, encoded by Chkb, results in minimal muscular dystrophy in forelimbs. We have found that choline kinase beta is the major isoform in hindlimb muscle and contributes more to choline kinase activity, while choline kinase alpha is predominant in forelimb muscle and contributes more to choline kinase activity. Although choline kinase activity is decreased in forelimb muscles of Chkb(-/-) mice, the activity of CTP:phosphocholine cytidylyltransferase is increased, resulting in enhanced phosphatidylcholine biosynthesis. The activity of phosphatidylcholine phospholipase C is up-regulated while the activity of phospholipase A(2) in forelimb muscle is not altered. Regeneration of forelimb muscles of Chkb(-/-) mice is normal when challenged with cardiotoxin. In contrast to hindlimb muscle, mega-mitochondria are not significantly formed in forelimb muscle of Chkb(-/-) mice. We conclude that the relative lack of muscle degeneration in forelimbs of Chkb(-/-) mice is due to abundant choline kinase alpha and the stable homeostasis of phosphatidylcholine. PMID- 20026285 TI - Fatty acid 2-Hydroxylation in mammalian sphingolipid biology. AB - 2-Hydroxy fatty acids (hFA) are important components of a subset of mammalian sphingolipids. The presence of hFA in sphingolipids is best described in the nervous system, epidermis, and kidney. However, the literature also indicates that various hFA-sphingolipids are present in additional tissues and cell types, as well as in tumors. Biosynthesis of hFA-sphingolipids requires fatty acid 2 hydroyxlase, and degradation of hFA-sphingolipids depends, at least in part, on lysosomal acid ceramidase and the peroxisomal fatty acid alpha-oxidation pathway. Mutations in the fatty acid 2-hydroxylase gene, FA2H, have been associated with leukodystrophy and spastic paraparesis in humans, underscoring the importance of hFA-sphingolipids in the nervous system. In the epidermis, hFA-ceramides are essential for the permeability barrier function. Physiological function of hFA sphingolipids in other organs remains largely unknown. Recent evidence indicates that hFA-sphingolipids have specific roles in cell signaling. PMID- 20026286 TI - The role of chemokines in recruitment of immune cells to the artery wall and adipose tissue. AB - The role of the immune system is to recognize pathogens, tumor cells or dead cells and to react with a very specific and localized response. By taking advantage of a highly sophisticated system of chemokines and chemokine receptors, leukocytes such as neutrophils, macrophages, and T-lymphocytes are targeted to the precise location of inflammation. While this is a beneficial process for acute infection and inflammation, recruitment of immune cells to sites of chronic inflammation can be detrimental. It is becoming clear that these inflammatory cells play a significant role in the initiation and progression of metabolic disorders such as atherosclerosis and insulin resistance by infiltrating the artery wall and adipose tissue (AT), respectively. Data from human studies indicate that elevated plasma levels of chemokines are correlated with these metabolic diseases. Recruitment of macrophages to the artery wall is well known to be one of the first steps in early atherosclerotic lesion formation. Likewise, recruitment of macrophages to AT is thought to contribute to insulin resistance associated with obesity. Based on this knowledge, much recent work in these areas has focused on the role of chemokines in attracting immune cells (monocytes/macrophages in particular) to these 2 sites. Thus, understanding the potential for chemokines to contribute to metabolic disease can help direct studies of chemokines as therapeutic targets. In this article, we will review current literature regarding the role of chemokines in atherosclerosis and obesity-related insulin resistance. We will focus on novel work showing that chemokine secretion from endothelial cells, platelets, and adipocytes can contribute to immune cell recruitment, with a diagram showing the time course of chemokine expression and leukocyte recruitment to AT. We will also highlight a few of the less-commonly known chemokine-chemokine receptor pairs. Finally, we will discuss the potential for chemokines as therapeutic targets for treatment of atherosclerosis and insulin resistance. PMID- 20026288 TI - Probing hydrogen peroxide oxidation kinetics of wild-type Synechocystis catalase peroxidase (KatG) and selected variants. AB - Catalase-peroxidases (KatGs) are unique bifunctional heme peroxidases that exhibit peroxidase and substantial catalase activities. Nevertheless, the reaction pathway of hydrogen peroxide dismutation, including the electronic structure of the redox intermediate that actually oxidizes H(2)O(2), is not clearly defined. Several mutant proteins with diminished overall catalase but wild-type-like peroxidase activity have been described in the last years. However, understanding of decrease in overall catalatic activity needs discrimination between reduction and oxidation reactions of hydrogen peroxide. Here, by using sequential-mixing stopped-flow spectroscopy, we have investigated the kinetics of the transition of KatG compound I (produced by peroxoacetic acid) to its ferric state by trapping the latter as cyanide complex. Apparent bimolecular rate constants (pH 6.5, 20 degrees C) for wild-type KatG and the variants Trp122Phe (lacks KatG-typical distal adduct), Asp152Ser (controls substrate access to the heme cavity) and Glu253Gln (channel entrance) are reported to be 1.2x10(4)M(-1)s(-1), 30M(-1)s(-1), 3.4x10(3)M(-1)s(-1), and 8.6x10(3)M(-1)s(-1), respectively. These findings are discussed with respect to steady-state kinetic data and proposed reaction mechanism(s) for KatG. Assets and drawbacks of the presented method are discussed. PMID- 20026289 TI - In vivo degradation of calcium phosphate cement incorporated into biodegradable microspheres. AB - In this study we have investigated the influence of the mechanism of microsphere degradation or erosion on the in vivo degradation of microsphere/calcium phosphate cement composites (microsphere CPCs) used in tissue engineering. Microspheres composed of poly(lactic-co-glycolic acid) (PLGA), gelatin and poly(trimethylene carbonate) (PTMC) were used as the model and the resulting microsphere CPCs were implanted subcutaneously for 4, 8 or 12weeks in the back of New Zealand white rabbits. Besides degradation, the soft tissue response to these formulations was evaluated. After retrieval, specimens were analyzed by physicochemical characterization and histological analysis. The results showed that all microsphere CPCs exhibited microsphere degradation after 12weeks of subcutaneous implantation, which was accompanied by decreasing compression strength. The PLGA microspheres exhibited bulk erosion simultaneously throughout the whole composite, whereas the gelatin type B microspheres were degradated from the outside to the center of the composite. High molecular weight PTMC microspheres exhibited surface erosion resulting in decreasing microsphere size. Furthermore, all composites showed a similar tissue response, with decreasing capsule thickness over time and a persistent moderate inflammatory response at the implant interface. In conclusion, microsphere CPCs can be used to generate porous scaffolds in an in vivo environment after degradation of microspheres by various degradation/erosion mechanisms. PMID- 20026287 TI - Glossopharyngeal long-term facilitation requires serotonin 5-HT2 and NMDA receptors in rats. AB - Although the glossopharyngeal nerve (IX) is mainly a sensory nerve, it innervates stylopharyngeus and some other pharyngeal muscles, whose excitations would likely improve upper airway patency since electrical IX stimulation increases pharyngeal airway size. As acute intermittent hypoxia (AIH) induces hypoglossal and genioglossal long-term facilitation (LTF), we hypothesized that AIH induces glossopharyngeal LTF, which requires serotonin 5-HT(2) and NMDA receptors. Integrated IX activity was recorded in anesthetized, vagotomized, paralyzed and ventilated rats before, during and after 5 episodes of 3-min isocapnic 12% O(2) with 3-min intervals of 50% O(2). Either saline, ketanserin (5-HT(2) antagonist, 2mg/kg) or MK-801 (NMDA antagonist, 0.2mg/kg) was (i.v.) injected 30-60 min before AIH. Both phasic and tonic IX activities were persistently increased (both P<0.05) after AIH in vehicle, but not ketanserin or MK-801, rats. Hypoxic glossopharyngeal responses were minimally changed after either drug. These data suggest that AIH induces both phasic and tonic glossopharyngeal LTF, which requires activation of 5-HT(2) and NMDA receptors. PMID- 20026290 TI - In vitro study of the proliferation and growth of human bone marrow cells on apatite-wollastonite-2M glass ceramics. AB - This study concerns the preparation and in vitro characterization of an apatite wollastonite-2M bioactive glass ceramic which is intended to be used for the regeneration of hard tissue (i.e. in dental and craniomaxillofacial surgery). This bioglass ceramic has been obtained by appropriate thermal treatment through the devitrification (crystallization) of a glass with a stoichiometric eutectic composition within the Ca(3)(PO(4))(2)-CaSiO(3) binary system. Crack-free specimens of the bioglass ceramic were immersed in human bone marrow cell cultures for 3, 7, 14 and 21days, in order to study biocompatibility. Cell morphology, proliferation and colonization were assessed by scanning electron microscopy and confocal laser scanning microscopy. A total protein content assay was used to evaluate the viability and proliferation of cultured bone marrow cells. The results showed that the cells were able to adhere and proliferate on the designed material due to the essentiality of silicon and calcium as accessory factors for cell activity stimulation. PMID- 20026291 TI - Transfer delay and in-hospital mortality of trauma patients in Pakistan. AB - BACKGROUND: Mortality and morbidity in trauma remain a major problem in developing countries. Organized emergency response systems for transfer of trauma patients to hospitals are absent and the consequent delays could cause significant complications. AIMS: This study assessed the outcomes as a result of hospital transfer and delays in trauma patients. METHODS: The study was based on trauma patients presenting to the Aga Khan University Hospital (AKUH), Karachi, Pakistan from 1998 to 2005, meeting the trauma team activation criteria. Data were collected and entered in a Trauma Registry. The study focused on analyzing the outcomes of injury to delay in definitive treatment and survival. RESULTS: Out of 978 patients, only 303 (30.9%) patients reached the emergency room (ER) within an hour. The mean time from injury occurrence to arrival in the ER was 4.7h. There was no significant difference in mortality between all patients presenting early and those with more than 1h delay (OR=0.9, 95% CI: 0.6, 1.5). CONCLUSIONS: Transfer and delay in admission to a tertiary care center does not affect in-hospital mortality of trauma patients in a setting with no emergency response system. This may be due to self selection of patients who survive long enough to reach the hospital. PMID- 20026292 TI - Quantitative proteomics: a central technology for systems biology. PMID- 20026293 TI - Quantification of the oxidative damage biomarker 2,3-dinor-8-isoprostaglandin F(2alpha) in human urine using liquid chromatography-tandem mass spectrometry. AB - F(2)-isoprostanes are useful biomarkers of oxidative status in humans. We developed an ultraperformance liquid chromatography-tandem mass spectrometric (UPLC-MS/MS) method to quantify 2,3-dinor-8-iso prostaglandin F(2alpha), a urinary metabolite of 8-iso-prostaglandin F(2alpha.) Urine was purified by solid phase extraction and analyzed by UPLC-MS/MS with negative-ion electrospray ionization. The method was robust with a mean inaccuracy of 9%, interday and intraday imprecision of 7.5% or lower, and a lower limit of quantification of 0.5 microg/L, equivalent to 0.04 pmol injected onto the column. An analysis time of 6 min was shorter than previously published methods and amenable to large studies. PMID- 20026294 TI - Anandamide levels in human female reproductive tissues: solid-phase extraction and measurement by ultraperformance liquid chromatography tandem mass spectrometry. AB - Anandamide (N-arachidonoylethanolamide), a bioactive lipid, is reported to play a role in pregnancy maintenance and parturition. Our aims were to (1) evaluate AEA levels at the human maternal:fetal interface and (2) validate the use of solid phase extraction of AEA from tissues. AEA was analyzed in cord and maternal blood, amniotic fluid, placenta, and fetal membranes collected during Caesarean section (n=14). Extraction efficiencies were 42 and 36% for the placenta and the fetal membranes, respectively. Tissue AEA was quantified using an isotope dilution method and UPLC-ESI-MS/MS giving intra- and inter-day variability for tissues spiked with 0.2, 1, and 5pmol/g AEA of less than 12%. Accuracy for these spiked samples was between 95% and 103% for fetal membranes and between 99% and 114% for placenta. Mean AEA concentrations were 2.72 + or - 1.04 pmol/g for placenta and 1.19 + or - 0.68 pmol/g for fetal membranes, and 0.93 + or - 0.28, 0.88 + or - 0.33, 0.77 + or - 0.30, and 0.06 + or - 0.04nM for maternal, umbilical vein, and umbilical artery plasma and amniotic fluid. Higher AEA concentrations were found in placenta compared to fetal membranes (P<0.0001), in umbilical vein compared with umbilical artery (P=0.0015), and in plasma from maternal circulation compared with umbilical artery (P=0.0152). The relevance of these changes in AEA concentrations at the maternal:fetal interface requires further investigation. PMID- 20026295 TI - Ultrasensitive detection of proteins and antibodies by absorption-based laser wave-mixing detection using a chromophore label. AB - Nonlinear laser wave mixing is presented as an ultrasensitive absorption-based method for the detection of proteins and antibodies using a nonfluorescing chromophore label, Coomassie Brilliant Blue (CBB). The complexes are flowed through a 150-microm (i.d.) capillary cell and detected using a low-power He-Ne laser. The wave-mixing signal is detected after 10 min of room temperature incubation for the antibody complex and after 18 min for the protein complex. All solutions are prepared in an aqueous buffer without the addition of organic modifiers. Concentration detection limits of 3.4 x 10(-19) and 6.4 x 10(-14) M (signal-to-noise ratio [S/N] = 2) are determined for bovine serum albumin (BSA) and human papillomavirus (HPV) antibody, respectively. Based on the small laser probe volume used (i.e., overlap volume of the two input beams), mass detection limits of 1.7 x 10(-22) and 2.6 x 10(-17) mol are determined for BSA and HPV antibody, respectively. PMID- 20026296 TI - Analysis of skeletal muscle metabolome: evaluation of extraction methods for targeted metabolite quantification using liquid chromatography tandem mass spectrometry. AB - Functional metabolomics of skeletal muscle involves the simultaneous identification and quantification of a large number of metabolites. For this purpose, the extraction of metabolites from animal tissues is a crucial technical step that needs to be optimized. In this work, five extraction methods for skeletal muscle metabolome analysis using liquid chromatography tandem mass spectrometry (LC-MS/MS) were tested. Bird skeletal muscles sampled postmortem and quenched in liquid nitrogen were used. Three replicates of the same sample were extracted using the following solvent systems of varying polarity: boiling water (BW, +100 degrees C), cold pure methanol (CPM, -80 degrees C), methanol/chloroform/water (MCW, -20 degrees C), boiling ethanol (BE, +80 degrees C), and perchloric acid (PCA, -20 degrees C). Three injections by extraction were performed. The BW extraction showed the highest recovery of metabolites with the lowest variability (<10%) except for creatine-phosphate (creatine-P). Considering yield (area of the peaks), reproducibility, and ease, the current experiment drew a scale for the muscle metabolome extraction starting from the best to the least convenient: BW>MCW>CPM>PCABE. In addition, the semiquantification of metabolites in two muscles showing different metabolic and contractile properties was carried out after BW extraction and showed expected differences in metabolite contents, thereby validating the technique for biological investigations. In conclusion, the BW extraction is recommended for analysis of skeletal muscle metabolome except for creatine-P, which was poorly recovered with this technique. PMID- 20026297 TI - Analysis of metabolomic PCA data using tree diagrams. AB - Large amounts of data from high-throughput metabolomic experiments are commonly visualized using a principal component analysis (PCA) two-dimensional scores plot. The question of the similarity or difference between multiple metabolic states then becomes a question of the degree of overlap between their respective data point clusters in principal component (PC) scores space. A qualitative visual inspection of the clustering pattern in PCA scores plots is a common protocol. This article describes the application of tree diagrams and bootstrapping techniques for an improved quantitative analysis of metabolic PCA data clustering. Our PCAtoTree program creates a distance matrix with 100 bootstrap steps that describes the separation of all clusters in a metabolic data set. Using accepted phylogenetic software, the distance matrix resulting from the various metabolic states is organized into a phylogenetic-like tree format, where bootstrap values 50 indicate a statistically relevant branch separation. PCAtoTree analysis of two previously published data sets demonstrates the improved resolution of metabolic state differences using tree diagrams. In addition, for metabolomic studies of large numbers of different metabolic states, the tree format provides a better description of similarities and differences between each metabolic state. The approach is also tolerant of sample size variations between different metabolic states. PMID- 20026299 TI - Interaction of the regulatory subunit of the cAMP-dependent protein kinase with PATZ1 (ZNF278). AB - The effects of cAMP in cell are predominantly mediated by the cAMP-dependent protein kinase (PKA), which is composed of two genetically distinct subunits, catalytic (C) and regulatory (R), forming a tetrameric holoenzyme R(2)C(2). The only known function for the R subunit is that of inhibiting the activity of the C subunit kinase. It has been shown that overexpression of RIalpha, but not the C subunit kinase, is associated with neoplastic transformation. In addition, it has also been demonstrated that mutation in the RIalpha, but not the C subunit is associated with increased resistance to the DNA-damaging anticancer drug cisplatin, thus suggesting that the RIalpha subunit of PKA may have functions independent of the kinase. We show here that the RIalpha subunit interacts with a BTB/POZ domain zinc-finger transcription factor, PATZ1 (ZNF278), and co expression with RIalpha results in its sequestration in the cytoplasm. The cytoplasmic/nuclear translocation is inducible by cAMP. C-terminus deletion abolishes PATZ1 interaction with RIalpha and results in its localization in the nucleus. PATZ1 transactivates the cMyc promoter and the presence of cAMP and co expression with RIalpha modulates its transactivation. Moreover, PATZ1 is aberrantly expressed in cancer. Taken together, our results showed a potentially novel mechanism of cAMP signaling mediated through the interaction of RIalpha with PATZ1 that is independent of the kinase activity of PKA, and the aberrant expression of PATZ1 in cancer point to its role in cell growth regulation. PMID- 20026300 TI - Inhibition of histone deacetylase activity is a novel function of the antifolate drug methotrexate. AB - Methotrexate (MTX) is a dihydrofolate reductase (DHFR) inhibitor widely used for treating human cancers, and overexpression of histone deacetylase (HDAC) is usually found in tumors. HDAC inhibitors (HDACi) can reactivate tumor suppressor genes and serve as potential anti-cancer drugs. In this study, we found that MTX shared structural similarity with some HDACi and molecular modeling showed that MTX indeed docks into the active site of HDLP, a bacterial homologue of HDAC. Subsequent in vitro assay demonstrated MTX's inhibition on HDAC activity in human cancer cells. The global acetylation of histone H3 was also induced by MTX. Moreover, MTX inhibited immunoprecipitated HDAC1/2 activity but not their protein levels. This study provides evidence that MTX inhibits HDAC activity. PMID- 20026298 TI - Overcoming rapid inactivation of lung surfactant: analogies between competitive adsorption and colloid stability. AB - Lung surfactant (LS) is a mixture of lipids and proteins that line the alveolar air-liquid interface, lowering the interfacial tension to levels that make breathing possible. In acute respiratory distress syndrome (ARDS), inactivation of LS is believed to play an important role in the development and severity of the disease. This review examines the competitive adsorption of LS and surface active contaminants, such as serum proteins, present in the alveolar fluids of ARDS patients, and how this competitive adsorption can cause normal amounts of otherwise normal LS to be ineffective in lowering the interfacial tension. LS and serum proteins compete for the air-water interface when both are present in solution either in the alveolar fluids or in a Langmuir trough. Equilibrium favors LS as it has the lower equilibrium surface pressure, but the smaller proteins are kinetically favored over multi-micron LS bilayer aggregates by faster diffusion. If albumin reaches the interface, it creates an energy barrier to subsequent LS adsorption that slows or prevents the adsorption of the necessary amounts of LS required to lower surface tension. This process can be understood in terms of classic colloid stability theory in which an energy barrier to diffusion stabilizes colloidal suspensions against aggregation. This analogy provides qualitative and quantitative predictions regarding the origin of surfactant inactivation. An important corollary is that any additive that promotes colloid coagulation, such as increased electrolyte concentration, multivalent ions, hydrophilic non-adsorbing polymers such as PEG, dextran, etc. added to LS, or polyelectrolytes such as chitosan, also promotes LS adsorption in the presence of serum proteins and helps reverse surfactant inactivation. The theory provides quantitative tools to determine the optimal concentration of these additives and suggests that multiple additives may have a synergistic effect. A variety of physical and chemical techniques including isotherms, fluorescence microscopy, electron microscopy and X-ray diffraction show that LS adsorption is enhanced by this mechanism without substantially altering the structure or properties of the LS monolayer. PMID- 20026301 TI - Cell adhesion property affected by cyclooxygenase and lipoxygenase: Opto-electric approach. AB - Expression of cyclooxygenases (COX) and lipoxygenases (LOX) has been linked to many pathophysiological phenotypes, including cell adhesion. However, many current approaches to measure cellular changes are performed only in a fixed-time point. Since cells dynamically move in conjunction with the cell matrix, there is a pressing need for dynamic or time-dependent methods for the investigation of cell properties. In the presented study, we used stable human colorectal cancer cell lines ectopically expressing COX-1, COX-2, and 15LOX-1, to investigate whether expression of COX-1, COX-2, or 15LOX-1 would affect cell adhesion using our opto-electric methodology. In a fixed-time point experiment, only COX-1- and COX-2-expressing cells enhanced phosphorylation of focal adhesion kinase, but all the transfected cells showed invasion activity. However, in a real-time experiment using opto-electric approaches, transmitted cellular morphology was much different with tight adhesion being shown in COX-2 expressing cells, as imaged by differential interference contrast microscopy (DICM) and interference reflection contrast microscopy (IRCM). Furthermore, micro-impedance measurements showed a continued increase in both resistance and reactance of COX- and LOX transfected cells, consistent with the imaging data. Our data indicate that both COX- and LOX-expressing cells have strong cell-to-cell and cell-to-substrate adhesions, and that cell imaging analysis with cell impedance data generates fully reliable results on cell adhesion measurement. PMID- 20026302 TI - Expression of the hGM-CSF in the silk glands of germline of gene-targeted silkworm. AB - To express human granulocyte-macrophage colony-stimulating factor (hGM-CSF) gene in the silk glands of transformation silkworm (Bombyx mori) based on gene targeting, two fragments from fibroin heavy chain gene (fib-H) of silkworm were cloned and sequenced. One fragment contains the 1st exon and its downstream 1st intron's partial sequence; and the other fragment contains the 1st intron's partial sequence and the 2nd exon's partial sequence. Then the two fragments, as homologous arm, were inserted into pSK to generate a gene-targeted vector, pSK-HL A3GFP-FLP-GM-CSF-FLPA-HR in which a gfp gene driven by A3 promoter and an hGM-CSF gene under the control of fibroin light chain (fib-L) promoter were included. The vector was transferred into the silkworm eggs using sperm-mediated gene transfer. After being screened for green fluorescent, the transformation silkworm was obtained, whose genome was verified by PCR and dot hybridization to confirm whether the target genes had been integrated into the silkworm genome. Furthermore, in the posterior silk glands of the G4 generation transformation silkworms, a specific band with the molecular weight of 22kDa could be detected by Western blotting with an antibody against hGM-CSF, and the expression level of the hGM-CSF estimated by ELISA was approximately 1.26ng per gram fresh posterior silk gland. PMID- 20026303 TI - Carabrol suppresses LPS-induced nitric oxide synthase expression by inactivation of p38 and JNK via inhibition of I-kappaBalpha degradation in RAW 264.7 cells. AB - Carabrol, isolated from Carpesium macrocephalum, showed anti-inflammatory potential in LPS-induced RAW 264.7 murine macrophages. In present study, carabrol demonstrated the inhibitory activity on pro-inflammatory cytokines such as IL 1beta, IL-6 and TNF-alpha. In addition, mRNA and protein levels of iNOS and COX-2 were reduced by carabrol. Molecular analysis revealed that these suppressive effects were correlated with the inactivation of p38 and JNK via inhibition of NF kappaB activation. Immunoblotting showed that carabrol suppressed LPS-induced degradation of I-kappaBalpha and decreased nuclear translocation of p65. Taken together, these results suggest that carabrol can be a modulator of pro inflammatory signal transduction pathway in RAW 264.7 cells. PMID- 20026304 TI - Amino acid deprivation induces CREBZF/Zhangfei expression via an AARE-like element in the promoter. AB - CREBZF (also called ZF or Zhangfei) is a basic region-leucine zipper transcription factor that has been implicated in the herpesvirus infection cycle and related cellular processes. Since ATF4 is known to play a key role in cellular responses to various ER stresses as well as amino acid deprivation, we sought to examine the potential involvement of CREBZF in the amino acid response (AAR). We found that the CREBZF protein was induced by amino acid deprivation in the canine MDCK cells. We subsequently cloned a canine CREBZF promoter region ( 1767bp to +1bp) that responds to amino acid limitation. Using deletion mapping and site-directed mutagenesis, we identified a 9-bp sequence 5'-ATTCACTCA-3' in the promoter (-1227 to -1219), deletion of which resulted in a complete loss of inducibility by amino acid deprivation. This sequence is similar to the known amino acid response elements (AAREs) found in other AAR-inducible genes, such as CHOP (C/EBP homologous protein, also known as GADD153). These results suggest that CREBZF may be an amino acid stress sensor. Considering the AARE-like sequence found in CREBZF and other similarities between CREBZF and CHOP, we postulate that CREBZF and CHOP may be two sensors that regulate different yet related signaling pathways governing the AAR. PMID- 20026305 TI - Active creatine kinase is present in matrix vesicles isolated from femurs of chicken embryo: Implications for bone mineralization. AB - Proteomic analysis of matrix vesicles (MVs) isolated from 17-day-old chicken embryo femurs revealed the presence of creatine kinase. In this report we identified the enzyme functionally and suggest that the enzyme may participate in the synthesis of ATP from ADP and phosphocreatine within the lumen of these organelles. Then, ATP is converted by nucleotide hydrolyzing enzymes such as Na(+), K(+)-ATPase, protein kinase C, or alkaline phosphatase to yield inorganic phosphate (P(i)), a substrate for mineralization. Alternatively, ATP can be hydrolyzed by a nucleoside triphosphate pyrophosphatase phosphodiesterase 1 producing inorganic pyrophosphate (PP(i)), a mineralization inhibitor. In addition, immunochemical evidence indicated that VDAC 2 is present in MVs that may serve as a transporter of nucleotides from the extracellular matrix. We discussed the implications of ATP production and hydrolysis by MVs as regulatory mechanisms for mineralization. PMID- 20026306 TI - Glucagon-like peptide-1 (GLP-1) inhibits advanced glycation end product (AGE) induced up-regulation of VCAM-1 mRNA levels in endothelial cells by suppressing AGE receptor (RAGE) expression. AB - Glucagon-like peptide-1 (GLP-1) is one of the incretins, a gut hormone secreted from L cells in the intestine in response to food intake. It has been proposed as a potential therapeutic target for the treatment of patients with type 2 diabetes. However, the direct effects of GLP-1 on vascular injury in diabetes are largely unknown. Since there is a growing body of evidence that advanced glycation end products (AGE) and their receptor RAGE axis plays an important role in vascular complications in diabetes, this study investigated whether and how GLP-1 blocked the deleterious effects of AGE on human umbilical vein endothelial cells (HUVEC). GLP-1 receptor (GLP-1R) was expressed in HUVEC. GLP-1 dose dependently inhibited RAGE gene expression in HUVEC, which was blocked by small interfering RNAs raised against GLP-1R. An analogue of cyclic AMP also decreased RAGE mRNA level in HUVEC. Further, GLP-1 decreased reactive oxygen species generation and subsequently reduced vascular cell adhesion molecule-1 mRNA levels in AGE-exposed HUVEC. Our present study suggests that GLP-1 directly acts on HUVEC via GLP-1R and it could work as an anti-inflammatory agent against AGE by reducing RAGE expression via activation of cyclic AMP pathways. PMID- 20026307 TI - Structural characterization reveals that viperin is a radical S-adenosyl-L methionine (SAM) enzyme. AB - Viperin is an interferon-inducible protein inhibiting many DNA and RNA viruses. It contains an N-terminal transmembrane helix, a highly conserved C-terminus and a middle region carrying a CX3CX2C motif, characteristic of radical S-adenosyl-l methionine (SAM) enzymes. So far no structural characterization has been reported and reconstitution of the [4Fe-4S] cluster in viperin all failed. Here, by dissecting the 361-residue human viperin into 12 fragments, followed by extensive CD and NMR characterization, Viperin (45-361) was identified to be soluble and structured in buffers. Most importantly, we have successfully reconstituted the [4Fe-4S] cluster in Viperin (45-361), thus providing the first experimental evidence confirming that viperin is indeed a radical SAM enzyme. Furthermore, the C-terminus Viperin (214-361) which is insoluble in buffers but again can be solubilized in salt-free water appears to be only partially folded. Our results thus imply that the radical SAM enzyme activity may play a key role in the broad antiviral actions of viperin. PMID- 20026308 TI - Distinct ONE-GC transduction modes and motifs of the odorants: Uroguanylin and CO(2). AB - In a subset of the olfactory sensory neurons ONE-GC($) membrane guanylate cyclase is a central component of two odorant-dependent cyclic GMP signaling pathways. These odorants are uroguanylin and CO(2). The present study was designed to decipher the biochemical and molecular differences between these two odorant signaling mechanisms. The study shows (1) in contrast to uroguanylin, CO(2) transduction mechanism is Ca(2+)-independent. (2) CO(2) transduction site, like that of uroguanylin-neurocalcin delta, resides in the core catalytic domain, aa 880-1028, of ONE-GC. (3) The site, however, does not overlap the signature neurocalcin delta signal transduction domain, (908)LSEPIE(913). Finally, (4) this study negates the prevailing concept that CO(2) uniquely signals ONE-GC activity (Sun et al. [19]; Guo et al. [21]). It demonstrates that it also signals the activation of photoreceptor membrane guanylate cyclase ROS-GC1. These results show an additional new transduction mechanism of the membrane guanylate cyclases and broaden our understanding of the molecular mechanisms by which different odorants using a single guanylate cyclase can regulate diverse cyclic GMP signaling pathways. PMID- 20026309 TI - Induction of apoptosis by thymoquinone in lymphoblastic leukemia Jurkat cells is mediated by a p73-dependent pathway which targets the epigenetic integrator UHRF1. AB - The salvage anti-tumoral pathway which implicates the p53-related p73 gene is not yet fully characterized. We therefore attempted to identify the up- and down stream events involved in the activation of the p73-dependent pro-apoptotic pathway, by focusing on the anti-apoptotic and epigenetic integrator UHRF1 which is essential for cell cycle progression. For this purpose, we analyzed the effects of a known anti-neoplastic drug, thymoquinone (TQ), on the p53-deficient acute lymphoblastic leukemia (ALL) Jurkat cell line. Our results showed that TQ inhibits the proliferation of Jurkat cells and induces G1 cell cycle arrest in a dose-dependent manner. Moreover, TQ treatment triggers programmed cell death, production of reactive oxygen species (ROS) and alteration of the mitochondrial membrane potential (DeltaPsim). TQ-induced apoptosis, confirmed by the presence of hypodiploid G0/G1 cells, is associated with a rapid and sharp re-expression of p73 and dose-dependent changes of the levels of caspase-3 cleaved subunits. These modifications are accompanied by a dramatic down-regulation of UHRF1 and two of its main partners, namely DNMT1 and HDAC1, which are all involved in the epigenetic code regulation. Knockdown of p73 expression restores UHRF1 expression, reactivates cell cycle progression and inhibits TQ-induced apoptosis. Altogether our results showed that TQ mediates its growth inhibitory effects on ALL p53-mutated cells via the activation of a p73-dependent mitochondrial and cell cycle checkpoint signaling pathway which subsequently targets UHRF1. PMID- 20026310 TI - Selective enhancement of main olfactory input to the medial amygdala by GnRH. AB - In male hamsters mating behavior is dependent on chemosensory input from the main olfactory and vomeronasal systems, whose central pathways contain cell bodies and fibers of gonadotropin-releasing hormone (GnRH) neurons. In sexually naive males, vomeronasal organ removal (VNX), but not main olfactory lesions, impairs mating behavior. Intracerebroventricular (i.c.v.)-GnRH restores mating in sexually naive VNX males and enhances medial amygdala (Me) immediate-early gene activation by chemosensory stimulation. In sexually experienced males, VNX does not impair mating and i.c.v.-GnRH suppresses Me activation. Thus, the main olfactory system is sufficient for mating in experienced-VNX males, but not in naive-VNX males. We investigated the possibility that GnRH enhances main olfactory input to the amygdala in naive-VNX males using i.c.v.-GnRH and pharmacological stimulation (bicuculline/D,L-homocysteic acid mixture) of the main olfactory bulb (MOB). In sexually naive intact males there was a robust increase of Fos protein expression in the anteroventral medial amygdala (MeAv) with MOB stimulation, but no effect of GnRH. There was no effect of stimulation or GnRH in posterodorsal medial amygdala (MePd). In naive-VNX animals, GnRH increased Fos in MeAv and MePv. Only combined MOB stimulation and i.c.v.-GnRH produced a significant increase in Fos in the dorsal (reproduction-related) portion of MeP (MePd). When the animals were sexually experienced before VNX, a condition in which GnRH does not enhance mating, i.c.v.-GnRH combined with MOB stimulation suppressed Fos expression in MePd. This suggests a more selective effect of GnRH on olfactory input in MePd than elsewhere in medial amygdala of VNX males. PMID- 20026312 TI - Influence of activity-induced axonal hypoexcitability on transmission of descending and segmental signals. AB - In this experiment, the changes in excitability of motor axons produced after natural activity were measured in nine healthy subjects using 1 min of maximal voluntary contractions (MVC) of the abductor digiti minimi (ADM) by studying the relationship between stimulus intensity applied to the ulnar nerve and the size of the ADM compound muscle action potential (CMAP). On cessation of the contraction, there was a prominent right-shift of the input-output curve: the intensity required to produce a control CMAP approximately 60% of maximum, generated a post-contraction response approximately 25% of maximum. Similar changes occurred in the input-output curves obtained by recording the ulnar nerve volley evoked by same test stimulus for CMAP. Motor-evoked potential (MEP) and F waves (and H-reflex in one subject) were recorded from ADM before and after 1 min of MVC. On cessation of contraction, the MEP input-output curves exhibited a significant right-shift: the stimulus required to evoke a pre-contraction maximum MEP ( approximately 60% of maximum CMAP) generated a post-contraction response approximately 65% of initial values. One minute of MVC produced similar decreases of F ( approximately 35%)- and H ( approximately 30%)-ADM responses. All responses recovered their control value in 15-20 min after the end of contraction. The almost identical depressive effect produced by 1 min of MVC on peripherally and centrally generated muscle responses suggests a common conditioning factor. These findings are discussed within the context of activity induced motor axonal hyperpolarizion. PMID- 20026311 TI - Genetic variance contributes to dopamine and opioid receptor antagonist-induced inhibition of intralipid (fat) intake in inbred and outbred mouse strains. AB - Preference for and intake of solid and emulsified fat (intralipid) solutions vary across different mouse strains. Fat intake in rodents is inhibited by dopamine and opioid receptor antagonists, but any variation in these responses as a function of genetic background is unknown. Therefore, the present study compared the ability of dopamine D1-like (SCH23390) and general opioid (naltrexone) receptor antagonism to alter intake of fat emulsions (intralipid) in mice. Two hour intakes of 5% intralipid were measured (5-120 min) in seven inbred (BALB/c, C57BL/6, C57BL/10, DBA/2, SJL, SWR, 129P3) and one outbred (CD-1) mouse strains following treatment with vehicle, SCH23390 (50-1600 nmol/kg, ip) and naltrexone (0.001-5 mg/kg, sc). SCH23390 significantly, dose-dependently and differentially reduced intralipid intake at all five (DBA/2, SWR, CD-1), four (SJL, C57BL/6), three (129P3) and one (C57BL/10) of the doses tested, but failed to affect intralipid intake in BALB/c mice. Naltrexone significantly, dose-dependently and differentially reduced intralipid intake at all four (DBA/2), three (SWR, SJL), two (CD-1, C57BL/10) and one (C57BL/6, 129P3) of the doses tested, and also failed to affect intralipid intake in BALB/cJ mice. SCH23390 and naltrexone were respectively 13.3-fold and 9.3-fold more potent in inhibiting intralipid intake in the most sensitive (DBA/2) relative to the least sensitive (BALB/c) mouse strains. A strong positive relationship (r=0.91) was observed for the abilities of SCH23390 and naltrexone to inhibit intralipid intake across strains. These findings indicate that dopaminergic and opioid signaling mechanisms differentially control intralipid intake across different mouse strains, suggesting important genetic and pharmacological interactions in the short-term control of rewarding and post-ingestive consequences of fat intake. PMID- 20026313 TI - Pravastatin treatment causes a shift in the balance of hippocampal neurotransmitter binding densities towards inhibition. AB - Since pravastatin, a HMG-CoA reductase inhibitor, has recently been shown to reduce infarct volumes and glutamate release in a rat model of ischemic stroke, the aim of the present study was to investigate whether this neuroprotective effect may be due to a modulation of excitatory and inhibitory neurotransmitter receptors. Therefore, Wistar rats were treated six times in 4 days with pravastatin or saline and allowed to survive for 6 hours or 5 days (n=10 per time point and group), respectively. Using quantitative receptor autoradiography, ligand binding densities of [(3)H]MK-801, [(3)H]AMPA, and [(3)H]muscimol for labeling of NMDA, AMPA, and GABA(A) receptors were analyzed in sensorimotor cortices Par1 and Par2, the striatum, and the hippocampus. Statin therapy induced complex alterations of ligand binding densities in different brain regions. Labeling of NMDA receptors was significantly increased in Par2, both after 6 hours and 5 days, respectively. Within the striatum, AMPA as well as GABA(A) receptor binding values were significantly increased on day 5. Furthermore, a marked and significant increase of [(3)H]muscimol ligand binding to GABA(A) receptors throughout all hippocampal subfields was seen after 6 hours. This complexity could easily be unraveled when focusing on the balance between excitatory glutamate and inhibitory GABA(A) receptors, in which case only the increase of hippocampal [(3)H]muscimol ligand binding 6 hours after the first application of pravastatin was accompanied by a net shift towards inhibition. Consequently, our data suggest an additional regulatory pathway induced by statins, namely modification of the abundance of excitatory and inhibitory neurotransmitter receptors. PMID- 20026314 TI - Clinical utility of copper, ceruloplasmin, and metallothionein plasma determinations in human neurodegenerative patients and their first-degree relatives. AB - The concentration of plasma copper, ceruloplasmin (CRP), non-ceruloplasmin-bound Cu (NCBC), and metallothioneins (MTs) were studied as putative biomarkers for neurodegenerative diseases in patients and in their first-degree relatives. We found increased levels of Cu in the plasma of Alzheimer's disease (AD), Parkinson's disease (PD), and vascular dementia (VD) patients, and the increase observed in VD group was linked to the evolution of the disease. CRP was also elevated in response to the inflammatory component of the diseases, however, a correlation with illness progression was only observed in VD patients. The level of MTs is proportional to the evolution of VD. The Cu/CRP and Cu/MTs ratios are both indicative of disease progression for AD patients but not for those with PD or VD. Moreover, there is a correlation between the NCBC levels and the cognitive impairment estimated through the Mini-mental State Examination (MMSE) scale. This dependence is linear for AD and PD patients and non-linear for the VD ones. The relative values of NCBC showed dependence on the disease duration, especially for AD. Copper measurement and the Cu/CRP ratio may be predictive markers of risk for the first-degree relatives of AD patients. We believe that these results are valuable as a reliable clinical tool. PMID- 20026315 TI - Subarachnoid hemorrhage induces enhanced expression of thromboxane A2 receptors in rat cerebral arteries. AB - Cerebral ischemia remains the key cause of morbidity and mortality after subarachnoid hemorrhage (SAH) with a pathogenesis that is still poorly understood. The aim of the present study was to examine the involvement of thromboxane A(2) receptors (TP) in the pathophysiology of cerebral ischemia after SAH in cerebral arteries. SAH was induced in rats by injecting 250 microl of blood into the prechiasmatic cistern. Two days after the SAH, cerebral arteries were harvested and contractile responses to the TP receptor agonist U46619 were investigated with myographs. In addition, the contractile responses were examined after pretreatment with selective TP receptor antagonist GR3219b. The TP receptor RNA and protein levels were analyzed by quantitative real-time PCR and immunohistochemistry, respectively. The global and regional cerebral blood flows (CBFs) were quantified with an autoradiographic technique. SAH resulted in enhanced contractile responses to U46619 as compared to sham. The TP receptor antagonist GR3219b abolished the enhanced contractile responses to U46619 observed after SAH. The TP receptor mRNA level was elevated after SAH as compared to sham. The level of TP receptor protein on the smooth muscle cells (SMCs) was increased in SAH compared to sham. Global and regional CBFs were reduced in SAH as compared to sham. The results demonstrate that SAH results in CBF reduction and this is associated with the enhanced expression of TP receptors in the SMC of cerebral arteries and microvessels. PMID- 20026316 TI - Rho kinase inhibition protects CA1 cells in organotypic hippocampal slices during in vitro ischemia. AB - The actin cytoskeleton is a dynamic superstructure that regulates multiple cellular functions and that has been implicated in cell death regulation. We investigated whether modulating the neuronal actin cytoskeleton polymerization by Rho-GTPase kinase (ROCK) inhibition influences cell death in hippocampal neuronal cultures and in murine organotypic hippocampal slice cultures subjected to in vitro ischemia (IVI). During IVI, spines on vehicle treated hippocampal neurons collapsed and large dendritic actin aggregates were formed. Following ROCK inhibition by Y27632, the actin aggregates were markedly smaller while large filopodia extended from the dendritic trunk. Y27632 also provided strong neuroprotection of hippocampal pyramidal CA1 neurons, which was of similar magnitude as protection by NMDA receptor blockade. Likewise, treatment with the F actin depolymerizing agent latrunculin during IVI diminished actin aggregation and mitigated cell death following IVI. We propose that ROCK inhibition protects neurons against ischemic damage by disrupting actin polymerization thereby mitigating NMDA receptor induced toxicity and releasing ATP bound to actin for cellular energy use. We conclude that ROCK inhibitors abrogate multiple detrimental processes and could therefore be useful in stroke therapy. PMID- 20026317 TI - Speaking words in two languages with one brain: neural overlap and dissociation. AB - The present study investigated the neural overlap and dissociation underlying overt word production in the first language (L1) and second language (L2). Twenty four Chinese-English bilinguals named pictures in either L1 or L2 while being scanned with functional magnetic resonance imaging (fMRI). When comparing picture naming in L2 to naming in L1, increased activity in the left inferior frontal gyrus, bilateral supplementary motor areas (SMA), left precentral gyrus, left lingual gyrus, left cuneus, bilateral putamen, bilateral globus pallidus, bilateral caudate and bilateral cerebellum were observed. This suggested that word production in L2 is less automatic and needs to recruit more neural resources for lexical retrieval, articulatory processing and cognitive control than in L1. In contrast, picture naming in L1 relative to picture naming in L2 revealed increased activity in the right putamen and right globus pallidus probably due to different phonological features between Chinese and English. In addition, the conjunction analysis, for the first time, revealed the common neural correlates underlying picture naming in L1 and L2. PMID- 20026318 TI - Pregnancy inhibits cell proliferation and neuroblast differentiation without neuronal damage in the hippocampal dentate gyrus in C57BL/6N mice. AB - Neural changes occur in the dam during gestation, and brain size has been shown to decrease across pregnancy in humans as well as rodents. In this study, we monitored neuronal damage, cell proliferation and neuroblast differentiation in the hippocampal dentate gyrus (DG) at age-matched virgin control (17- to 18-week old), gestation day (GD) 14.5, 16.5 and 18.5 (17- to 18-week-old dams), using NeuN for mature neurons, terminal deoxynucleotidyl dUTP nick-end labeling (TUNEL) and Fluoro-Jade B (F-J B) for neuronal death, Ki67 for cell proliferation and doublecortin (DCX) for neuroblast differentiation in C57BL/6 mice. There were no significant differences in NeuN-immunoreactive ((+)) neurons between the age matched control and gestating groups. TUNEL or F-J B positive neurons were rarely detected in the DG in all the groups. Ki67(+) cell proliferation was significantly decreased in the subgranular zone of the dentate gyrus (SZDG) at GD16.5. In addition, DCX(+) neuroblasts with/without tertiary dendrites were decreased in the SZDG with gestation by GD16.5. However, in the GD18.5 group, the number of Ki67(+) nuclei and DCX(+) neuroblasts with/without tertiary dendrites was slightly increased compared to that observed at GD16.5. DCX protein levels were low at GD16.5, and thereafter slightly increased. These results suggest that cell proliferation and neuroblast differentiation in DG of the hippocampus is decreased during gestation. PMID- 20026319 TI - Inhibitory effects of (2S, 3S)-3-[3-[4 (trifluoromethyl)benzoylamino]benzyloxy]aspartate (TFB-TBOA) on the astrocytic sodium responses to glutamate. AB - Astrocytes are responsible for the majority of the clearance of extracellular glutamate released during neuronal activity. dl-threo-beta-benzyloxyaspartate (TBOA) is extensively used as inhibitor of glutamate transport activity, but suffers from relatively low affinity for the transporter. Here, we characterized the effects of (2S, 3S)-3-[3-[4-(trifluoromethyl)benzoylamino]benzyloxy]aspartate (TFB-TBOA), a recently developed inhibitor of the glutamate transporter on mouse cortical astrocytes in primary culture. The glial Na(+)-glutamate transport system is very efficient and its activation by glutamate causes rapid intracellular Na(+) concentration (Na(+)(i)) changes that enable real time monitoring of transporter activity. Na(+)(i) was monitored by fluorescence microscopy in single astrocytes using the fluorescent Na(+)-sensitive probe sodium-binding benzofuran isophtalate. When applied alone, TFB-TBOA, at a concentration of 1 microM, caused small alterations of Na(+)(i). TFB-TBOA inhibited the Na(+)(i) response evoked by 200 microM glutamate in a concentration dependent manner with IC(50) value of 43+/-9 nM, as measured on the amplitude of the Na(+)(i) response. The maximum inhibition of glutamate-evoked Na(+)(i) increase by TFB-TBOA was >80%, but was only partly reversible. The residual response persisted in the presence of the AMPA/kainate receptor antagonist CNQX. TFB-TBOA also efficiently inhibited Na(+)(i) elevations caused by the application of d-aspartate, a transporter substrate that does not activate non-NMDA ionotropic receptors. TFB-TBOA was found not to influence the membrane properties of cultured cortical neurons recorded in whole-cell patch clamp. Thus, TFB-TBOA, with its high potency and its apparent lack of neuronal effects, appears to be one of the most useful pharmacological tools available so far for studying glial glutamate transporters. PMID- 20026320 TI - Resting hippocampal blood flow, spatial memory and aging. AB - Aging is accompanied by a general deterioration of fluid cognitive processes and a reduction in resting cerebral blood flow (CBF). While the two phenomena have been observed independently, it is uncertain whether individual differences in cerebral blood flow are reliably associated with cognitive functioning in older adults. Furthermore, previous studies have concentrated primarily on gross measures of cognition and global gray matter CBF, leaving open the possibility that perfusion of specific brain regions may relate differentially to distinct cognitive domains. The present study sought to provide a more focused treatment of CBF and cognitive function in the context of aging by investigating the relationships among aging, spatial memory and resting hippocampal blood flow, both between and within younger and older adult groups. Blood flow was quantified using a novel Flow-Enhanced Signal Intensity (FENSI) technique which provides a localized, functionally relevant measure of volumetric flow across a given unit area. As expected, we found that aging was associated with poorer spatial memory and reduced resting CBF. Moreover, hippocampal blood flow was positively correlated with spatial memory performance in the older adult group, suggesting that increased blood flow to the hippocampus is associated with superior memory performance in older adults. These results demonstrate a region-specific CBF cognition relationship and thereby offer new insight into the complex connection between the aging brain and behavior. PMID- 20026321 TI - Human benzene metabolism following occupational and environmental exposures. AB - We previously reported evidence that humans metabolize benzene via two enzymes, including a hitherto unrecognized high-affinity enzyme that was responsible for an estimated 73% of total urinary metabolites [sum of phenol (PH), hydroquinone (HQ), catechol (CA), E,E-muconic acid (MA), and S-phenylmercapturic acid (SPMA)] in nonsmoking females exposed to benzene at sub-saturating (ppb) air concentrations. Here, we used the same Michaelis-Menten-like kinetic models to individually analyze urinary levels of PH, HQ, CA and MA from 263 nonsmoking Chinese women (179 benzene-exposed workers and 84 control workers) with estimated benzene air concentrations ranging from less than 0.001-299 ppm. One model depicted benzene metabolism as a single enzymatic process (1-enzyme model) and the other as two enzymatic processes which competed for access to benzene (2 enzyme model). We evaluated model fits based upon the difference in values of Akaike's Information Criterion (DeltaAIC), and we gauged the weights of evidence favoring the two models based upon the associated Akaike weights and Evidence Ratios. For each metabolite, the 2-enzyme model provided a better fit than the 1 enzyme model with DeltaAIC values decreasing in the order 9.511 for MA, 7.379 for PH, 1.417 for CA, and 0.193 for HQ. The corresponding weights of evidence favoring the 2-enzyme model (Evidence Ratios) were: 116.2:1 for MA, 40.0:1 for PH, 2.0:1 for CA and 1.1:1 for HQ. These results indicate that our earlier findings from models of total metabolites were driven largely by MA, representing the ring-opening pathway, and by PH, representing the ring-hydroxylation pathway. The predicted percentage of benzene metabolized by the putative high-affinity enzyme at an air concentration of 0.001 ppm was 88% based upon urinary MA and was 80% based upon urinary PH. As benzene concentrations increased, the respective percentages of benzene metabolized to MA and PH by the high-affinity enzyme decreased successively to 66 and 77% at 0.1 ppm, 20 and 58% at 1 ppm, and 2.7 and 17% at 10 ppm. This indicates that the putative high-affinity enzyme was active primarily below 1 ppm and favored the ring-opening pathway. PMID- 20026322 TI - A hospital-based case control study of aplastic anemia in Shanghai, China. AB - We report results of a hospital-based case control study of 137 consecutive patients diagnosed with aplastic anemia (AA) in participating hospitals over a 4 year period. Diagnoses were made by a single laboratory, subjects were age- and gender-matched to two controls and interviewed concerning previous disease, work histories and exposures to potential etiologic agents. Analysis was conducted on two distinct subgroups: severe aplastic anemia (SAA) and moderate aplastic anemia (MAA). In univariate regression models, the strongest associations were observed for exposure to benzene and SAA (OR=3.12, 95% CI=1.12-8.65) and life on a farm and MAA (OR=3.08, 95% CI=1.44-6.56). Benzene exposure did not show a strong dose response relationship with either subtype. When accounting for all of the potential confounders we considered in conditional regression models, the previous relationships persisted. Other explanatory variables included hair-dye use for MAA and farm exposures, such as livestock for SAA, although most of these additional variables fell just short of statistical significance. Adjusted R squared values were only 10% for each subtype, leaving 90% of AA occurrence unexplained. Our results suggest that: (a) benzene exposure is more strongly related to SAA than MAA, (b) farm and livestock exposures are related to both forms of AA, confirming some previous results, and (c) a large percentage of AA remains unexplained, which may indicate that individual susceptibility has a major influence on AA occurrence. PMID- 20026323 TI - Enhanced levels of soluble and membrane-bound CD137 levels in patients with acute coronary syndromes. AB - BACKGROUND: Increasing evidence shows that costimulatory molecules of the tumor necrosis factor superfamily such as CD40/CD40 ligand and OX40/OX40 ligand have been implicated in atherosclerosis. We investigated whether the expression levels of the tumor necrosis factor superfamily members CD137 in serum and membrane bound were related to acute coronary syndromes (ACS). METHODS: Thirty normal controls and 210 patients, including 70 with stable angina (SA), 80 with unstable angina (UA), and 60 with acute myocardial infarction (AMI), were enrolled in our study. The expression of CD137 in peripheral monocytes was analyzed by flow cytometry. Serum soluble CD137 (sCD137) and C-reactive protein levels were measured by commercially available ELISA. RESULTS: The expression of CD137 in peripheral monocytes in patients with UA [14.2+/-3.5 mean fluorescence intensity (MFI)] and AMI (15.1+/-4.4MFI) was significantly higher than those in patients with SA (6.5+/-2.4MFI) and controls (7.1+/-3.5MFI). sCD137 in patients with UA (16.7+/-4.9 ng/ml) and AMI (19.1+/-4.3 ng/ml) were significantly higher than those in patients with SA (3.4+/-1.4 ng/ml) and controls (3.9+/-1. 3 ng/ml) (p<0.001). C-reactive protein level in serum in patients with UA (13.8+/-3.3 ng/ml) and AMI (15.5+/-4.7 ng/ml) were also higher than those in patients with SA (1.4+/-0.4 ng/ml) and controls (1.3+/-0.3ng/ml). It was interesting that percutaneous transluminal coronary angioplasty (PTCA) induced a marked rise in sCD137 levels in SA patients, while CD137 expression in peripheral monocytes showed no difference between SA patients with PTCA before and after. A positive correlation was found between sCD137 and serum C-reactive protein levels (r=0.681; p<0.0001). CONCLUSION: Patients with ACS showed increased soluble and membrane-bound CD137 expression. sCD137 level showed a significantly positive correlation with CRP level in patients with ACS. The relation between sCD137 and ACS needs further researches. PMID- 20026324 TI - Methionine sulfoxide reductase B2 is highly expressed in the retina and protects retinal pigmented epithelium cells from oxidative damage. AB - Methionine sulfoxide reductase B2 (MSRB2) is a mitochondrial enzyme that converts methionine sulfoxide (R) enantiomer back to methionine. This enzyme is suspected of functioning to protect mitochondrial proteins from oxidative damage. In this study we report that the retina is one of the human tissues with highest levels of MSRB2 mRNA expression. Other tissues with high expression were heart, kidney and skeletal muscle. Overexpression of an MSRB2-GFP fusion protein increased the MSR enzymatic activity three-fold in stably transfected cultured RPE cells. This overexpression augmented the resistance of these cells to the toxicity induced by 7-ketocholesterol, tert-butyl hydroperoxide and all-trans retinoic acid. By contrast, knockdown of MSRB2 by a miRNA in stably transfected cells did not convey increased sensitivity to the oxidative stress. In the monkey retina MSRB2 localized to the ganglion cell layer (GLC), the outer plexiform layer (OPL) and the retinal pigment epithelium (RPE). MSRB2 expression is most pronounced in the OPL of the macula and foveal regions suggesting an association with the cone synaptic mitochondria. Our data suggests that MSRB2 plays an important function in protecting cones from multiple type of oxidative stress and may be critical in preserving central vision. PMID- 20026325 TI - Curcumin protects against hyperosmoticity-induced IL-1beta elevation in human corneal epithelial cell via MAPK pathways. AB - Increased tear osmolarity is an essential feature of dry eye disease. Curcumin, a natural polyphenol extracted from herb turmeric, has recently been reported to have anti-inflammatory effects. However, its anti-inflammatory effects have not been investigated in dry eye disease. It has been reported that elevated osmolarity achieved by adding sodium chloride to the culture medium of corneal epithelial cells increased the production of IL-1beta, a proinflammation cytokine. This in vitro dry eye model was used to test the anti-inflammatory effects of curcumin. In the present study, a 450 mOsM hyperosmotic medium was produced by adding sodium chloride to the culture medium to reach a final concentration of 90mM. Human corneal epithelial cells cultured in this hyperosmotic medium for 24h showed an increase of IL-1beta, IL-6 and TNF-alpha levels in the conditioned medium. IL-1beta was also upregulated at mRNA levels. Activation of p38 MAP kinase (p38), JNK MAP kinase (JNK) and NF-kappaB in cultured corneal epithelial cells were also induced by hyperosmotic conditions. Curcumin at concentrations of 1-30muM did not affect the cell viability of cultured corneal epithelial cells. Pretreatment of curcumin (5muM) completely abolished the increased production of IL-1beta induced by the hyperosmotic medium. Increased phosphorylation of p38 caused by high osmolarity was also completely abolished by curcumin, whereas the phosphorylation of JNK was only partially inhibited. SB 203580 (p38 inhibitor), but not SP 600125 (JNK inhibitor), completely suppressed hyperosmoticity-induced IL-1beta production, indicating that the inhibition of production of IL-1beta by curcumin may be achieved through the p38 signal pathway. Curcumin completely abolished a hyperosmoticity-induced increase of NF-kappaB p65. NF-kappaB inhibitor suppressed hyperosmoticity-induced IL-1beta production. p38 inhibitor suppressed hyperosmoticity-induced NF-kappaB activation, indicating that NF-kappaB activation was dependent on p38 activation. The present study suggests that curcumin might have therapeutic potential for treating dry eye disease. PMID- 20026326 TI - Panky, a novel photoreceptor-specific ankyrin repeat protein, is a transcriptional cofactor that suppresses CRX-regulated photoreceptor genes. AB - Neuronal gene transcription is regulated by both transcriptional activators and repressors. While the roles of transactivators in retinal photoreceptor development have been well characterized, the roles of repressors have been poorly understood. We isolated Panky/Ankrd33, a gene encoding an ankyrin repeat containing protein. Panky-A was specifically expressed in retinal photoreceptors and the pineal gland, and its expression was directly up-regulated by the CRX transcription factor. Subcellular localization of PANKY-A was observed in the nucleus and cytoplasm. Additionally, transactivation analysis suggested that PANKY-A is a transcriptional cofactor that suppresses CRX-activated photoreceptor genes. Furthermore, we found by an electrophoretic mobility shift assay that PANKY inhibited the DNA-binding activity of CRX. PMID- 20026327 TI - Methyl-Typing: an improved and visualized COBRA software for epigenomic studies. AB - Combined bisulfite restriction analysis (COBRA) is one of the most commonly used methylation quantification methods. However, it focuses on relatively few restriction enzymes. Here, we present Methyl-Typing, a web-based software that provides restriction enzyme mining data for methyl-cytosine-containing sequences following bisulfite-conversion. Gene names, accession numbers, sequences, PCR primers, and file upload are accessible for input. Promoter sequences and restriction enzymes for CpG- and GpC-containing recognition sites are retrieved. Four representative enzymes were tested successfully by COBRA on the experimental work. Therefore, the Methyl-Typing tool provides a comprehensive COBRA restriction enzyme mining. It is freely available at http://bio.kuas.edu.tw/methyl-typing. PMID- 20026328 TI - The yeast aquaglyceroporin Fps1p is a bidirectional arsenite channel. AB - The stress-activated kinase Hog1p mediates arsenic tolerance by decreasing arsenite influx through the aquaglyceroporin Fps1p in Saccharomyces cerevisiae. Unexpectedly, we found that overexpression of FPS1 increased arsenite tolerance suggesting a physiological role of Fps1p in arsenic detoxification. Consistently, during arsenite treatment transcription of FPS1 gene was strongly upregulated, while Fps1p was not degraded and remained localized to the plasma membrane. Moreover, deletion of FPS1 gene resulted in arsenate sensitivity. Finally, transport experiments revealed that Fps1p in concert with the arsenite transporter Acr3p mediates arsenite efflux. PMID- 20026329 TI - Gentamicin inhibits HSP70-assisted protein folding by interfering with substrate recognition. AB - We previously reported that gentamicin (GM) specifically binds to heat-shock protein with subunit molecular masses of 70 kDa (HSP70). In the present study, we have investigated the effects of GM binding on HSP70-assisted protein folding in vitro. The C-terminal, and not the N-terminal of HSP70 was found to bind to GM. GM significantly suppressed refolding of firefly luciferase in the presence of HSP70 and HSP40, although the ATPase activity of HSP70 was unaffected by GM. A surface plasmon resonance analysis revealed that GM specifically interferes with the binding of HSP70 to a model peptide that mimics the exposed hydrophobic surface of the folding intermediates. These results indicated that GM inhibits the chaperone activity of HSP70 and may suppress protein folding via inhibition of HSP70 in vivo. PMID- 20026330 TI - Chaperone-mediated autophagy in health and disease. AB - Chaperone-mediated autophagy (CMA) is a lysosomal pathway that participates in the degradation of cytosolic proteins. CMA is activated by starvation and in response to stressors that result in protein damage. The selectivity intrinsic to CMA allows for removal of damaged proteins without disturbing nearby functional ones. CMA works in a coordinated manner with other autophagic pathways, which can compensate for each other. Interest in CMA has recently grown because of the connections established between this autophagic pathway and human pathologies. Here we review the unique properties of CMA compared to other autophagic pathways and its relevance in health and disease. PMID- 20026331 TI - Divalent metal-dependent regulation of hepcidin expression by MTF-1. AB - Hepcidin is a small acute phase peptide that regulates iron absorption. It is induced by inflammation and infection, but is repressed by anaemia and hypoxia. Here we further reveal that hepcidin transcription also involves interactions between functional metal response elements (MREs) in its promoter, and the MRE binding transcription factor-1. Analysis of hepcidin mRNA and protein levels in hepatoma cells suggests that its expression may be regulated by divalent metal ions, with zinc inducing maximal effects on hepcidin levels. These data suggest that this peptide may be a pleiotropic sensor of divalent metals, some of which are xenobiotic environmental toxins. PMID- 20026332 TI - Disulfide linkage in the coiled-coil domain of subunit H of A1AO ATP synthase from Methanocaldococcus jannaschii and the NMR structure of the C-terminal segment H(85-104). AB - The C-terminal residues 98-104 are important for structure stability of subunit H of A(1)A(O) ATP synthases as well as its interaction with subunit A. Here we determined the structure of the segment H(85-104) of H from Methanocaldococcus jannaschii, showing a helix between residues Lys90 to Glu100 and flexible tails at both ends. The helix-helix arrangement in the C-terminus was investigated by exchange of hydrophobic residues to single cysteine in mutants of the entire subunit H (H(I93C), H(L96C) and H(L98C)). Together with the surface charge distribution of H(85-104), these results shine light into the A-H assembly of this enzyme. PMID- 20026333 TI - Positive selection on apoptosis related genes. AB - Apoptosis is a form of programmed cell death crucial for development, homeostasis, immunity, spermatogenesis, and prevention of cancer. Positive selection acting on mammalian apoptosis related genes targets protein interfaces that interact with pathogens and also elements of signaling complexes. Selection appears primarily to be driven by the immune/defense related function of these genes. Moreover, competitive interactions could be driving positive selection among sperm cells, as well as the need for protection against female anti-sperm immune responses. Trade-offs in fitness are expected out of these selective pressures, which could explain the involvement of these genes in various diseases, including cancer. PMID- 20026334 TI - Annual profile of fecal androgen and glucocorticoid levels in free-living male American kestrels from southern mid-latitude areas. AB - Fecal samples and behavioral data were collected at a fortnightly basis during 11 months period from free-living male American kestrels living in southeast Brazil (22 degrees S latitude). The aim was to investigate the seasonal changes in testicular and adrenal steroidogenic activity and their correlation to reproductive behaviors and environmental factors. The results revealed that monthly mean of fecal glucocorticoid metabolites in May and June were higher than those estimated in November. In parallel, monthly mean of androgen metabolites in September was higher than those from January to April and from October to November. Molt took place from January to March, whereas copulation was observed from June to October but peaked in September. Nest activity and food transfer to females occurred predominantly in October, and parental behavior was noticed only in November. Territorial aggressions were rare and scattered throughout the year. Multiple regression analysis revealed that fecal androgen levels are predicted by photoperiod and copulation, while fecal glucocorticoid levels are only predicted by photoperiod. Bivariate correlations showed that fecal androgen metabolites were positively correlated with fecal glucocorticoid metabolites and copulation, but negatively correlated with molt. Additionally, copulation was positively correlated with food transfer to females and nest activity, but negatively correlated with molt. These findings suggest that male American kestrels living in southeast Brazil exhibit significant seasonal changes in fecal androgen and glucocorticoid concentrations, which seem to be stimulated by decreasing daylength but not by rainfall or temperature. PMID- 20026335 TI - Crustacean hyperglycemic hormone (CHH) neuropeptidesfamily: Functions, titer, and binding to target tissues. AB - The removal of the eyestalk (s) induces molting and reproduction promoted the presence of regulatory substances in the eyestalk (ES), particularly medulla terminalis X-organ and the sinus gland (MTXO-SG). The PCR-based cloning strategies have allowed for isolating a great number of cDNAs sequences of crustacean hyperglycemic hormone (CHH) neuropeptides family from the eyestalk and non-eyestalk tissues, e.g., pericardial organs and fore- and hindguts. However, the translated corresponding neuropeptides in these tissues, their circulating concentrations, the mode of actions, and specific physiological functions have not been well described. The profiles of CHH neuropeptides present in the MTXO-SG may differ among decapod crustacean species, but they can be largely divided into two sub-groups on the basis of structural homology: (1) CHH and (2) molt inhibiting hormone (MIH)/mandibular organ-inhibiting hormone (MOIH)/vitellogenesis/gonad-inhibiting hormone (V/GIH). CHH typically elevating the level of circulating glucose from animals under stressful conditions (hyper- and hypothermia, hypoxia, and low salinity) has multiple target tissues and functions such as ecdysteroidogenesis, osmoregulation, and vitellogenesis. Recently, MIH, known for exclusively suppressing ecdysteroidogenesis in Y-organs, is also reported to have an additional role in vitellogenesis of adult female crustacean species, suggesting that some CHH neuropeptides may acquire an extra regulatory role in reproduction at adult stage. This paper reviews the regulatory roles of CHH and MIH at the levels of specific functions, temporal and spatial expression, titers, their binding sites on the target tissues, and second messengers from two crab species: the blue crab, Callinectes sapidus, and the European green crab, Carcinus maenas. It further discusses the diverse regulatory roles of these neuropeptides and the functional plasticity of these neuropeptides in regard to life stage and species-specific physiology. PMID- 20026336 TI - Two modular forms of the mitochondrial sorting and assembly machinery are involved in biogenesis of alpha-helical outer membrane proteins. AB - The mitochondrial outer membrane contains two translocase machineries for precursor proteins--the translocase of the outer membrane (TOM complex) and the sorting and assembly machinery (SAM complex). The TOM complex functions as the main mitochondrial entry gate for nuclear-encoded proteins, whereas the SAM complex was identified according to its function in the biogenesis of beta-barrel proteins of the outer membrane. The SAM complex is required for the assembly of precursors of the TOM complex, including not only the beta-barrel protein Tom40 but also a subset of alpha-helical subunits. While the interaction of beta-barrel proteins with the SAM complex has been studied in detail, little is known about the interaction between the SAM complex and alpha-helical precursor proteins. We report that the SAM is not static but that the SAM core complex can associate with different partner proteins to form two large SAM complexes with different functions in the biogenesis of alpha-helical Tom proteins. We found that a subcomplex of TOM, Tom5-Tom40, associates with the SAM core complex to form a new large SAM complex. This SAM-Tom5/Tom40 complex binds the alpha-helical precursor of Tom6 after the precursor has been inserted into the outer membrane in an Mim1 (mitochondrial import protein 1)-dependent manner. The second large SAM complex, SAM-Mdm10 (mitochondrial distribution and morphology protein), binds the alpha helical precursor of Tom22 and promotes its membrane integration. We suggest that the modular composition of the SAM complex provides a flexible platform to integrate the sorting pathways of different precursor proteins and to promote their assembly into oligomeric complexes. PMID- 20026337 TI - A molecular ruler mechanism for length control of extended protein structures in bacteria. AB - The lengths of the hook structure of flagellar motors and of the needle of the injectosome are both carefully controlled, by apparently similar mechanisms. In this paper we propose a novel mechanism for this length control and develop a mathematical model of this process which shows excellent agreement with published data on hook lengths. The proposed mechanism for length control (described using biochemical nomenclature appropriate for hooks) is as follows: Hook growth is terminated when the C-terminus of the length control molecule FliK interacts with FlhB, the secretion gatekeeper. The probability of this interaction is an increasing function of the length of the hook for two reasons. First, FliK is secreted through the hook intermittently during hook growth. Second, the probability of interaction with FlhB is a function of the amount of time the C terminus of a secreted FliK spends in the vicinity of FlhB. This time is short when the hook is short because the folding of FliK exiting the distal end of the hook acts to pull the FliK molecule through the hook rapidly. In contrast, this time is much longer when the hook is longer than the unfolded FliK polymer since movement through the tube is not enhanced by folding. Thus, it is much more likely that interaction will occur when the hook is long than when the hook is short. PMID- 20026338 TI - Effects of density dependent sex allocation on the dynamics of a simultaneous hermaphroditic population: modelling and analysis. AB - In this work we present a mathematical model describing the dynamics of a population where sex allocation remains flexible throughout adult life and so can be adjusted to current environmental conditions. We consider that the fractions of immature individuals acquiring male and female sexual roles are density dependent through nonlinear functions of a weighted total population size. The main goal of this work is to understand the role of life-history parameters on the stabilization or destabilization of the population dynamics. The model turns out to be a nonlinear discrete model which is analysed by studying the existence of fixed points as well as their stability conditions in terms of model parameters. The existence of more complex asymptotic behaviours of system solutions is shown by means of numerical simulations. Females have larger fertility rate than males. On the other hand, increasing population density favours immature individuals adopting the male role. A positive equilibrium of the system exists whenever fertility and survival rates of one of the sexual roles, if shared by all adults, allow population growing while the opposite happens with the other sexual role. In terms of the female inherent net reproductive number, eta(F), it is shown that the positive equilibria are stable when eta(F) is larger and closed to 1 while for larger values of eta(F) a certain asymptotic assumption on the investment rate in the female function implies that the population density is permanent. Depending on the other parameters values, the asymptotic behaviour of solutions becomes more complex, even chaotic. In this setting the stabilization/destabilization effects of the abruptness rate in density dependence, of the survival rates and of the competition coefficients are analysed. PMID- 20026339 TI - Elevated expression of semaphorin 5A in human gastric cancer and its implication in carcinogenesis. AB - AIMS: Semaphorin 5A, a member of semaphorin family, was originally identified as axonal guidance factor functioning during neuronal development. Here, we investigated semaphorin 5A expression in gastric cancer and explored its roles in gastric carcinogenesis. MAIN METHODS: The expression of semaphorin 5A was examined by reverse transcription-polymerase chain reaction (RT-PCR) analysis in six gastric cancer cell lines and detected by real-time RT-PCR and Western blotting in 30 pairs of primary gastric cancer and normal gastric mucosa tissues. RNA interference (RNAi) technique was used to generate a semaphorin 5A-silenced stable cell line. 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and anchorage-independent growth as well as flow cytometry assays were conducted to examine the effect of siRNA-semaphorin 5A on gastric cancer cell growth, proliferation and apoptosis. KEY FINDINGS: Semaphorin 5A was expressed in all human gastric cancer lines and the expression of semaphorin 5A was significantly higher in cancer tissues than that in normal mucosa tissues. siRNA mediated semaphorin 5A knockdown significantly suppressed the proliferation and anchorage-independent growth, and induced the apoptosis of gastric cancer cell line SGC7901. SIGNIFICANCE: The present study suggests that overexpression of semaphorin 5A may contribute to gastric carcinogenesis, which reveals a novel expression and function of semaphorin 5A outside the nervous system and adds more weight to our knowledge of semaphorin 5A. PMID- 20026340 TI - Relationship between heterochromatic interstitial telomeric sequences and chromosome damage induced by the radiomimetic compound streptonigrin in Chinese hamster ovary cells. AB - The relationship between (heterochromatic) interstitial telomeric sequences (ITSs) and the chromosome damage induced by the radiomimetic compound streptonigrin (SN) was investigated in Chinese hamster ovary (CHO) cells by using PNA- and Q-FISH techniques with a pantelomeric probe. CHO cells were exposed to increasing concentrations of SN and chromosomal aberrations were analyzed in the first mitosis after treatment. Cytogenetic analysis revealed that 16.9% and 11.7% of the total aberrations induced by SN in cells harvested 18 h and 3 h after treatment, respectively, exhibited one or more FISH-detectable telomeric signals. Although there was a significant induction by SN of chromosome breaks at centromeric regions containing ITSs, about 70% of the chromosome breaks exhibiting telomeric signals observed in SN-treated cells occurred outside the centromeric regions of chromosomes. This observation, along with the finding of entirely labeled acentric fragments in both untreated and SN-treated cells show that, although this antibiotic induces breakage at centromeric regions containing ITSs, these chromosome regions are not the preferential target for the clastogenic action of SN. In addition, our results show that heterochromatic ITSs are involved more than expected in the formation of chromatid breaks and exchanges induced by SN, and that these sequences are not preferentially involved in the formation of dicentrics, multicentrics, centric rings, chromosome breaks, acentric fragments and chromatid deletions induced by this antibiotic. These findings indicate that the involvement of heterochromatic ITSs in the chromosome damage induced by SN is not random. Moreover, our results show that SN induces telomeric repeats translocations, although this effect depends on the concentration of the drug, and that this antibiotic increases the size of ITSs, this latter effect not being related to the chromosomal sensitivity of the exposed cells to this compound. The mechanism by which SN induces amplification of heterochromatic ITSs remains to be elucidated. PMID- 20026341 TI - Nitric oxide mediated DNA double strand breaks induced in proliferating bystander cells after alpha-particle irradiation. AB - Low-dose alpha-particle exposures comprise 55% of the environmental dose to the human population and have been shown to induce bystander responses. Previous studies showed that bystander effect could induce stimulated cell growth or genotoxicity, such as excessive DNA double strand breaks (DSBs), micronuclei (MN), mutation and decreased cell viability, in the bystander cell population. In the present study, the stimulated cell growth, detected with flow cytometry (FCM), and the increased MN and DSB, detected with p53 binding protein 1 (53BP1) immunofluorescence, were observed simultaneously in the bystander cell population, which were co-cultured with cells irradiated by low-dose alpha particles (1-10 cGy) in a mixed system. Further studies indicated that nitric oxide (NO) and transforming growth factor beta1 (TGF-beta1) played very important roles in mediating cell proliferation and inducing MN and DSB in the bystander population through treatments with NO scavenger and TGF-beta1 antibody. Low concentrations of NO, generated by spermidine, were proved to induce cell proliferation, DSB and MN simultaneously. The proliferation or shortened cell cycle in bystander cells gave them insufficient time to repair DSBs. The increased cell division might increase the probability of carcinogenesis in bystander cells since cell proliferation increased the probability of mutation from the mis-repaired or un-repaired DSBs. PMID- 20026343 TI - Electrophysiological evidence of enhanced cortical activity in the human brain during visual curve tracing. AB - ERPs were recorded while participants performed a curve tracing task in which they had to identify the end point of a target curve presented among three other distractor curves. Differential activation associated with the side of the target curve was found in the form of a sustained posterior contralateral negativity (SPCN). This contralateral brain activity suggests covert attention was deployed to the target curve during performance of the tracing task. The amplitude of the SPCN varied according to the hypothesized curve-tracing process, depending on whether the start and end locations of the target curve were above to below the horizontal midline, or the opposite, and this detailed analysis of the results provided evidence supporting the spread-of-attention model of curve tracing. These results represent the first neurophysiological investigation of brain activity reflecting visual curve tracing in humans. PMID- 20026342 TI - The relevance of the functional 5-HT1A receptor polymorphism for attention and working memory processes during mental rotation of characters. AB - Numerous lines of research indicate that attentional processes, working memory and saccadic processes are highly interrelated. In the current study, we examine the relation between these processes with respect to their cognitive neurophysiological and neurobiological background by means of event-related potentials (ERPs) in a sample of N=72 healthy probands characterized for the functional serotonin receptor 1 A (5-HT1A) C(-1019)G polymorphism. The results support a close interrelation between working memory, attentional and saccadic processes. Yet, these processes are differentially modulated by the 5-HT1A C( 1019)G polymorphism. The 5-HT1A C(-1019)G polymorphism primarily affects attentional processing, whereas processes related to the mental rotation of an object are independent of 5-HT1A genetic variation. It is shown that an increasing number of -1019 G alleles leads to a differential reduction of the N1 above the left and right hemisphere and hence bottom-up attentional processing. In the way increasing numbers of -1019 G alleles lead to a reduction of attentional processes, saccadic activity increases as a similar function of the number of -1019 G alleles. This increase in activity occurs parallel in time to the process of mental rotation. It is hypothesized that decreased attentional processes, dependent on different 5-HT1A C(-1019)G genotypes, may cause parietal networks to increase saccadic activity in order to perform mental rotation. PMID- 20026344 TI - Calorie restriction causes healthy life span extension in the filamentous fungus Podospora anserina. AB - Although most fungi appear to be immortal, some show systemic senescence within a distinct time frame. Podospora anserina for example shows an irreversible growth arrest within weeks of culturing associated with a destabilization of the mitochondrial genome. Here, we show that calorie restriction (CR), a regimen of under-nutrition without malnutrition, increases not only life span but also forestalls the aging-related decline in fertility. Similar to respiratory chain deficiencies the life span extension is associated with lower levels of intracellular H(2)O(2) measurements and a stabilization of the mitochondrial genome. Unlike respiratory chain deficiencies, CR cultures have a wild-type-like OXPHOS machinery similar to that of well-fed cultures as shown by native electrophoresis of mitochondrial protein complexes. Together, these data indicate that life span extension via CR is fundamentally different from that via respiratory chain mutations: Whereas the latter can be seen as a pathology, the former promotes healthy life span extension and may be an adaptive response. PMID- 20026345 TI - Solubilization and structural determination of a glycoconjugate which is assembled into the sheath of Leptothrix cholodnii. AB - The sheath of Leptothrix cholodnii is constructed from a structural glycoconjugate, a straight-chained amphoteric heteropolysaccharide modified with glycine and cysteine. Though the structure of the glycan core is already determined, its modifications with amino acids and other molecules are not fully resolved. In this study, we aimed to determine the chemical structure of the glycoconjugate as a whole. Enantiomeric determination of cysteine in the sheath was performed and as a result, L-cysteine was detected. NMR spectroscopy was endeavored to determine overall structure of the glycoconjugate. Prior to NMR analysis, solubilization of the glycoconjugate was attempted by adding denaturing reagents or by derivatization. As far as tested, sulfonation by performic acid oxidation was suitable for solubilization, but further improvement was achieved by N-acetylation. The approximate molecular weight of the derivative was estimated to be 4.5 x 10(4) by size-exclusion chromatography. The NMR studies for the sulfonated glycoconjugate and its N-acetylated derivative revealed that the sheath glycoconjugate is a glycosaminoglycan consisting of a pentasaccharide repeating unit which is substoichiometrically esterified with 3-hydroxypropionic acid and stoichiometrically amidated with acetic acid and glycyl-L-cysteine. PMID- 20026346 TI - Sulfation of tea polysaccharides: synthesis, characterization and hypoglycemic activity. AB - Neutral polysaccharides (NTPS) and acid polysaccharides (ATPS) from tea leaves were obtained on a D315 macroporous anion-exchange resin column chromatography. NTPS and ATPS were sulfated by the pyridine-sulfonic acid method to obtain NTPS-S and ATPS-S. It was found that NTPS was easier sulfated than ATPS. There are strong characteristic absorption peaks located in 1258 cm(-1), 1146 cm(-1), 832 cm(-1) and 617 cm(-1) in the FTIR spectra of sulfated polysaccharides. Sulfation of polysaccharides also affected the endothermic and exothermic peaks via the DSC scan analysis. The appearance of exothermic peaks in both NTPS-S and ATPS-S indicated that the redox reaction might happen. The comparative study of hypoglycemic effect on mice showed that the sulfation of polysaccharides significantly improved hypoglycemic activity. PMID- 20026347 TI - Isolation and characterization of chicken lung mesenchymal stromal cells and their susceptibility to avian influenza virus. AB - In this study, we isolated and characterized mesenchymal stromal cells (MSCs) from the lungs of 1- to 2-week-old chickens. Microscopically, the cultured cells showed fibroblast-like morphology. Phenotypically these cells expressed CD44, CD90, CD105 and the transcription factor PouV, which has been shown to be critical for stem cell self-renewal and pluripotency. The multipotency of chicken MSCs was demonstrated by their ability to undergo adipogenic and osteogenic differentiation. Like chicken bone marrow MSCs and mammalian MSCs, chicken lung MSCs had immunoregulatory activity and profoundly suppressed the proliferative capacity of T cells in response to a mitogenic stimulus. Next, we examined the susceptibility of these cells to H1N1 and H9N5 avian influenza (AI) viruses. The lung MSCs were shown to express known influenza virus alpha-2,3 and alpha-2,6 sialic acid receptors and to support replication of both the avian H1N1 and avian H9N5 influenza strains. Viral infection of MSCs resulted in cell lysis and cytokine and chemokine production. Further characterization of lung MSCs in chicken and other mammalian species may help in understanding the pathogenesis of infectious and non-infectious lung diseases and the mechanisms of lung injury repair. PMID- 20026348 TI - In search of the origin of FREPs: characterization of Aplysia californica fibrinogen-related proteins. AB - All haemolymph lectins with uniquely juxtaposed N-terminal domain similar to the immunoglobulin superfamily (IgSF) and C-terminal fibrinogen (FBG) termed FBG related proteins (FREP) are documented till now only in the pulmonate mollusc Biomphalaria glabrata. Using genomic WGS database we have found two FREP genes from marine opistobranch Aplysia californica named AcFREP1 and AcFREP2. The AcFREP1 and AcFREP2 mRNA molecules have been subsequently isolated from cDNA of sea hare larvae as well as adult mollusc tissues. These genes encode proteins (504 and 510aa respectively) with domain architecture typical for FREPs with two N-terminal IgSF domains and C-terminal FBG domain. Although cDNA sequences of AcFREP1 and AcFREP2 are 81% identical, their genomic structure is entirely different: AcFREP1 is intronless and AcFREP2 is encoded in four exons. These genes are paralogous pair in which AcFREP2 is a parental gene and AcFREP1 is the new transposed copy that has lost the introns (retrogene). Using RT-PCR analysis, expression of AcFREP1 and AcFREP2 was shown to be developmentally and tissue specific and no constitutive expression in haemocytes was found. The overall frequency of nucleotide substitutions in genomic DNA trace sequences of coding region of the AcFREP1 and AcFREP2 is not higher than in the sequences of control conserved genes (actin, FMRFamide). Thus, previously reported high diversification of Biomphalaria FREP gene, BgFREP3, is not detected in Aplysia FREPs. A search for FREP homologs in other available complete genome of mollusc, Lottia gigantea (Patellogastropoda), a representative of the evolutionary earliest gastropod clade, did not reveal any DNA sequences coding for similar lectins. We suggest that unique domain architecture of FREPs is an evolutionary novelty that appeared and evolved only within one branch of Protostomata species, exclusively in heterobranch molluscs (Pulmonata and Opistobranchia). PMID- 20026349 TI - Inflammation as a psychophysiological biomarker in chronic psychosocial stress. AB - The measurement of inflammation by biomarkers not only documents clinically relevant infections but also offers an important tool to pin point potentially harmful effects of chronic psychosocial stressors. This article focuses firstly on basic biology of inflammation and lists main biomarkers currently used in psycho-physiologic research. In the second part, the effects of the hypothalamic pituitary-adrenal (HPA) axis and the autonomic nervous system as pathways modulating stress-related inflammation are discussed. Furthermore, current evidence of how chronic psychosocial stressors are related to alterations in inflammatory activity is presented. In summary, job stress, low socioeconomic status, childhood adversities as well as life events, caregiver stress, and loneliness were all shown to exert effects on immunologic activity. PMID- 20026350 TI - The cortisol awakening response: more than a measure of HPA axis function. AB - In most healthy people morning awakening is associated with a burst of cortisol secretion: the cortisol awakening response (CAR). It is argued that the CAR is subject to a range physiological regulatory influences that facilitate this rapid increase in cortisol secretion. Evidence is presented for reduced adrenal sensitivity to rising levels of ACTH in the pre-awakening period, mediated by an extra-pituitary pathway to the adrenal from the suprachiasmatic nucleus (SCN). A role for the hippocampus in this pre-awakening regulation of cortisol secretion is considered. Attainment of consciousness is associated with 'flip-flop' switching of regional brain activation, which, it is argued, initiates a combination of processes: (1) activation of the hypothalamic pituitary adrenal (HPA) axis; (2) release of pre-awakening reduced adrenal sensitivity to ACTH; (3) increased post-awakening adrenal sensitivity to ACTH in response to light, mediated by a SCN extra-pituitary pathway. An association between the CAR and the ending of sleep inertia is discussed. PMID- 20026351 TI - Sensorimotor adaptation of saccadic eye movements. AB - Sensory-motor adaptation mechanisms play a pivotal role in maintaining the performance of goal-directed movements. The saccadic system, used to explore the visual environment through fast and accurate shifts of the eyes (saccades), is a valuable model for studying adaptation mechanisms. Significant progresses have been recently made in identifying the properties and neural substrates of saccadic adaptation elicited by the double-step target paradigm. Behavioural data collected in healthy and brain-damaged subjects, and neurophysiological data from non human primates, will be reviewed in an attempt to build a coherent picture of saccadic adaptation mechanisms. Emphasis will further be put on the contextual factors of saccadic adaptation, and on the link between adaptive changes of oculomotor commands and visual perception. It will be shown that saccadic adaptation relies on multiple mechanisms according to experimental contexts, time scales, saccade categories, and direction of adaptive changes of saccade amplitude (shortening versus lengthening). Taking into account this complexity will be a key toward a comprehensive understanding of the physiopathology of saccadic adaptation and toward the development of possible rehabilitation procedures. PMID- 20026352 TI - Recurrent bacteraemia: A 10-year regional population-based study of clinical and microbiological risk factors. AB - BACKGROUND: A population-based nested case-control study was conducted in order to characterize patient factors and microbial species associated with recurrent bacteraemia. METHODS: All patients with bacteraemia in a Danish region during 1996-2006 were investigated. Recurrence was defined based on pathogen identity, site of infection and time frame, and not restricted to homologous pathogens. RESULTS: We identified 8672 patients with first-time bacteraemia, of whom 1003 (12%) had a recurrence within 1 year. The proportion of mono-microbial bacteraemia was similar for first (86%) and recurrent episodes (84%). An unknown focus was common in both episodes (22.7 and 29.1%, respectively). Independent predictors of a recurrence (incidence rate ratio, 95% confidence interval) included health care-associated (2.4; 1.9-3.0) and nosocomial bacteraemia (2.1; 1.8-2.6), poly-microbial Gram-positive bacteraemia (2.7; 1.6-4.6), and fungaemia (2.2; 1.4-3.5), a Charlson co-morbidity index score of 1-2 (1.7; 1.4-2.1), inappropriate empirical antimicrobial chemotherapy (1.3; 1.1-1.5), a gastro intestinal tract focus (2.3; 1.7-3.0), a liver/biliary tract focus (2.7; 2.0 3.6), an iv-catheter focus (2.0; 1.4-2.8), endocarditis (2.7; 1.6-4.3), and an unknown focus (1.9; 1.5-2.3). CONCLUSIONS: This study showed recurrent bacteraemia to be common and the following risk factors were identified: a health care-associated or nosocomial origin, poly-microbial or fungal aetiology, a focus within the abdomen, endocardium, iv-catheter-related or unknown, a Charlson co morbidity index score of >1 and inappropriate empirical antimicrobial chemotherapy. PMID- 20026353 TI - Improving evaluations of T-cell assays for diagnosing active Mycobacterium tuberculosis infection. PMID- 20026354 TI - Neuronal and non-neuronal GABA transporters as targets for antiepileptic drugs. AB - Epileptic seizure activity is associated with an imbalance between excitatory and inhibitory synaptic activities. The latter is mediated by GABA, and several currently used antiepileptic drugs target entities of the GABAergic synapse such as the receptors or the inactivation mechanism consisting of transmembrane transport and enzymatic degradation. The development of tiagabine selectively inhibiting the GABA transporter GAT1 constitutes a proof of concept that the GABA transporters are interesting drug targets in the context of antiepileptic drugs. The review provides a detailed analysis of the role of such transporters pointing in particular to an interesting role of the transporters located extrasynaptically. It is suggested that the betaine-GABA transporter BGT1 should receive particular interest in this context as the GABA analogue EF 1502 (N-[4,4 bis(3-methyl-2-thienyl)-3-butenyl]-4-(methylamino)-4,5,6,7 tetrahydrobenzo[d]isoxazol-3-ol) has been shown to possess a novel anticonvulsant profile in animal models of epilepsy, involving the ability to inhibit GABA transport mediated by GAT1 and BGT1 at the same time. PMID- 20026355 TI - Peroxisome proliferator-activated receptor beta/delta (PPARbeta/delta) acts as regulator of metabolism linked to multiple cellular functions. AB - Peroxisome proliferator-activated receptors (PPARs) are nuclear receptors. They function as ligand activated transcription factors. They exist in three isoforms, PPARalpha, PPARbeta (formerly PPARdelta), and PPARgamma. For all PPARs lipids are endogenous ligands, linking them directly to metabolism. PPARs form heterodimers with retinoic X receptors, and, upon ligand binding, modulate gene expression of downstream target genes dependent on the presence of co-repressors or co activators. This results in cell-type specific complex regulations of proliferation, differentiation and cell survival. Specific synthetic agonists for all PPARs are available. PPARalpha and PPARgamma agonists are already in clinical use for the treatment of hyperlipidemia and type 2 diabetes, respectively. More recently, PPARbeta activation came into focus as an interesting novel approach for the treatment of metabolic syndrome and associated cardiovascular diseases. Although the initial notion was that PPARbeta is expressed ubiquitously, more recently extensive investigations have been performed demonstrating high PPARbeta expression in a variety of tissues, e.g. skin, skeletal muscle, adipose tissue, inflammatory cells, heart, and various types of cancer. In addition, in vitro and in vivo studies using specific PPARbeta agonists, tissue-specific over-expression or knockout mouse models have demonstrated a variety of functions of PPARbeta in adipose tissue, muscle, skin, inflammation, and cancer. We will focus here on functions of PPARbeta in adipose tissue, skeletal muscle, heart, angiogenesis and cancer related to modifications in metabolism and the identified underlying molecular mechanisms. PMID- 20026356 TI - Voltage-gated calcium channels in the etiopathogenesis and treatment of absence epilepsy. AB - Voltage-gated calcium channels are key elements in regulating neuronal excitability and are thus of central importance in the pathogenesis of various forms of epilepsies. Among these, absence epilepsies represent about 10% of epileptic seizures in humans. They are electroencephalographically characterized by bilateral synchronous spike-wave discharge activity associated with loss or severe impairment of consciousness. Extensive studies during the last decades revealed that pathophysiologically increased oscillatory activity, i.e., hyperoscillation within the reticulothalamocortical circuitry, is the electrophysiological correlate of absence epilepsy, with extrathalamocortical structures, e.g., brainstem and cerebellum, projecting to the thalamocortical circuitry, thereby modulating its activity. Voltage-gated calcium channels are one of the central players regulating the transition from tonic to rebound burst firing modes in both thalamic relay and reticular thalamic nucleus neurons, the burst-firing mode being the substrate of the thalamocortical oscillation. Thus, pharmacological interference with these channels enables effective control of spike-wave discharge activity in patients suffering from absence seizures. In this review, we summarize the medical history of absence epilepsies, their classification and terminology, the diagnostic armamentarium available today and the etiopathogenesis of absences. Finally, various antiepileptic drugs that have been proven to or are supposed to exert anti-absence effects are discussed with respect to their pharmacodynamics and pharmacokinetics. PMID- 20026357 TI - Optimized cannula design and placement for convection-enhanced delivery in rat striatum. AB - The stereotactic delivery of therapeutic agents into brain has been problematic because of reflux and leakage of the delivered agent. Good distribution of infusates by convection-enhanced delivery (CED) depends very much on cannula design, precise cannula placement and infusion rates. We have recently published cannula targeting data for the non-human primate (NHP) putamen in which we defined infusion parameters referred to as "red", "blue", and "green" zones for cannula placements that result in poor, sub-optimal and optimal volumes of distribution (Vd), respectively. Therefore, we applied our observations in NHP putamen to the rat brain. Initially, trypan blue dye was infused into agarose gels to evaluate distribution and reflux characteristics of a scaled-down cannula without step and 1-mm stepped cannula. "Stepped" means a sharp transition from a wider stent to a narrower tip; thus the distance of the cannula tip to the larger diameter attachment defines the step distance. Reflux was contained with the stepped design even with an infusion rate of 3.0 microl/min and large infusion volumes in the agarose gel study. Infusions of a recombinant growth factor, GDNF, into rat striatum demonstrated that the presence of a 1-mm stepped cannula prevented reflux and resulted in excellent distribution of GDNF in the striatum. We conclude that a stepped cannula with a 1-mm tip is important for achieving reliable distribution of infused agents in rat brain. It should be considered when local therapies such as gene transfer, local protein administration or cellular replacement are evaluated in rodent models. PMID- 20026358 TI - Rafts and the battleships of defense: the multifaceted microdomains for positive and negative signals in immune cells. AB - Recognition of the heterogeneity of the cell membrane was one of the most important scientific achievements in the last decades. Since coining the term "lipid rafts", continuous development of advanced microscopic and spectroscopic techniques has vastly expanded our view on these cell membrane microdomains that appear to have almost as many faces as researchers that look at them; they are variable in stability, size and composition that can change in a highly dynamic manner both by recruiting and expelling components as well as by coalescing and breaking up into smaller units. They have, however, one common feature: all eukaryotic cells present some variation of lipid rafts. Cells of the immune system are not exception to this, regardless of their lymphoid or myeloid origin their membranes show a domain structure and these domains serve to condense or reject particular transmembrane, GPI-linked and intracellularly membrane-anchored proteins as function requires. Here we provide a concise overview about the various weapons and shields that immune cells concentrate into their rafts, which have come into sight during the past years. The positive and negative regulatory roles of these microdomains are essential both in the functions of innate immunity and processes concatenated in the adaptive immune response. PMID- 20026360 TI - Meprin-alpha metalloproteases enhance lipopolysaccharide-stimulated production of tumour necrosis factor-alpha and interleukin-1beta in peripheral blood mononuclear cells via activation of NF-kappaB. AB - Lipopolysaccharide (LPS) induces the expression of a wide range of pro inflammatory mediators via NF-kappaB activation. These pro-inflammatory cytokines, such as tumor necrosis factor-alpha (TNF-alpha) and interleukin-1beta (IL-1beta), may be important in triggering atherogenesis. We have previously observed that actinonin, a meprin inhibitor, suppressed the formation of atherosclerotic plaques and, in in vitro experiments, actinonin also had an effect on the way LPS altered THP-1 cell function. The aim of the present study was to investigate whether meprin-alpha regulates LPS-induced production of TNF alpha and IL-1beta in peripheral blood mononuclear cells (PBMCs) and its potential mechanisms of action. We observed that meprin-alpha could enhance LPS induced expression of TNF-alpha and IL-1beta mRNA and protein in a time- and concentration-dependent manner, assessed using real-time PCR and ELISA. Meprin alpha also significantly increased LPS-induced NF-kappaB transcriptional activity. Furthermore, we assessed the effects of meprin-alpha specific siRNA on the production of TNF-alpha and IL-1beta to examine whether meprin-alpha was involved in the process of LPS-induced activation of PBMCs. Our results show that LPS-induced IL-1beta and TNF-alpha production by PBMCs was significantly reversed by meprin-alpha specific siRNA. In addition, the augmentation of meprin-alpha of the LPS-induced expression of TNF-alpha and IL-1beta was significantly decreased by Bay-117082, an inhibitor of NF-kappaB. In conclusion, our data indicate that meprin-alpha is capable of increasing LPS-induced production of cytokines in peripheral blood mononuclear cells, which might be associated with the activation of NF-kappaB. PMID- 20026361 TI - Oral delivery of mouse [D-Leu-4]-OB3, a synthetic peptide amide with leptin-like activity, in male Swiss Webster mice: a study comparing the pharmacokinetics of oral delivery to intraperitoneal, subcutaneous, intramuscular, and intranasal administration. AB - We have recently shown that intranasal administration of mouse [D-Leu-4]-OB3 reconstituted in Intravail to male Swiss Webster mice resulted in significantly higher bioavailability than commonly used injection methods of delivery. The absorption profile associated with intranasal delivery of mouse [D-Leu-4]-OB3 showed an early peak representing rapid uptake across the nasal mucosa, and a later peak suggesting a gastrointestinal site of absorption. In the present study, we show that gastrointestinal absorption of mouse [D-Leu-4]-OB3 does occur, and that reformulation of mouse [D-Leu-4-OB3 with Intravail significantly enhances its uptake. The pharmacokinetics of orally delivered (by gavage) mouse [D-Leu-4]-OB3 in the absence or presence of Intravail were examined, and compared to previously reported pharmacokinetic parameters of mouse [D-Leu-4]-OB3 following intraperitoneal (ip), subcutaneous (sc), intramuscular (im), and intranasal administration. When compared to oral delivery in PBS, Intravai significantly enhanced the total uptake (552,710 ng/ml/min vs.137,585 ng/ml/min) and relative bioavailability (4.0 vs. 1.0) of mouse [D-Leu-4-OB3. The relative oral bioavailabilities of mouse [D-Leu-4]-OB3 when compared to ip, sc, im, and intranasal delivery were 52.2%, 47.3%, 37.8% and 12.9%, respectively. The results of this study indicate that oral delivery of mouse [D-Leu-4]-OB3 in Intravail is an effective method of administration achieving relatively high serum levels of the bioactive peptide when compared to commonly used methods of injection. In addition to intranasal administration, oral delivery of mouse [D-Leu-4]-OB3 in Intravail may have potential as a novel, non-invasive approach to the treatment of obesity and its associated metabolic dysfunctions in humans. PMID- 20026359 TI - Phospholipid and sphingolipid metabolism in Leishmania. AB - In many eukaryotes, phospholipids (PLs) and sphingolipids (SLs) are abundant membrane components and reservoirs for important signaling molecules. In Leishmania, the composition, metabolism, and function of PLs and SLs differ significantly from those in mammalian cells. Although only a handful of enzymes have been experimentally characterized, available data suggest many steps of PL/SL metabolism are critical for Leishmania viability and/or virulence, and could be a source for new drug targets. Further studies of genes involved in the synthesis (de novo and salvage) and degradation of PLs and SLs will reveal their diverse effects on Leishmania pathogenesis. PMID- 20026363 TI - Fluorometric detection and estimation of fungal biomass on cultural heritage materials. AB - A wide variety of cultural heritage materials are susceptible to fungal deterioration. The paper, canvas, and stone constituents of our cultural heritage are subjected to harmful physical and chemical processes as they are slowly consumed by fungi. Remediation of fungal contamination can be costly and risk further damage to cultural artifacts. Early detection of fungal growth would permit the use of relatively noninvasive treatments to remediate fungal contamination before visible or lasting damage to the object has occurred. Current methods used for the detection and measurement of microbial biomass, such as colony counts, microscopic biovolume estimation, and ergosterol analysis are expensive and time consuming, or are inappropriate for use with fungi. Beta-N acetylhexosaminidase (3.2.1.52) activity provides a reliable estimation of fungal biomass in soil and on building materials. Adapted for use on cultural heritage materials' fluorogenic 4-methylumbelliferyl (MUF) labeled substrate N-acetyl-beta d-glucosaminide (NAG) was used to detect beta-N-acetylhexosaminidase activity in the fungus Aspergillus niger. Fluorescence increased linearly with fungal biomass and the sensitivity of the assay was comparable to other biochemical techniques. The fluorometric assay was used to monitor fungal biomass on a variety of cultural heritage materials non-destructively, and without the introduction of chemicals or solvents to the surfaces. PMID- 20026362 TI - Novel sample preparation method for molecular detection of Mollicutes in cell culture samples. AB - Research laboratories, raw materials and media suppliers as well as the biopharmaceutical industry face recurrent contamination with Mollicutes. Culture based detection methods are very slow (28 days) and could ideally be replaced by nucleic acid testing (NAT) for rapid result. These methods are nonetheless hampered by their companion sample preparation methods. They are limited by the volume tested (0.1 to 5 mL), the protein/nucleic acid content they can accommodate and are generally performed in an open environment. The processing of low volumes of complex matrices is associated to several issues such as poor representativeness, low sensitivity, inhibition and false positives. The novel sample preparation method described in this study has been developed to overcome these limitations and to process 20-mL samples containing high loads of eukaryotic cells. A dual-membrane device is coupled to magnetic bead purification. In one single and closed device, eukaryotic cells and microorganisms are separated, contaminants are concentrated, lysed and corresponding nucleic acids are collected. This novel sample preparation method has been tested with 9 different Mollicutes. The ability to detect the contaminants down to 0.6 CFU/mL by real-time PCR among hundreds of millions of CHO-S cells (Chinese hamster ovary cells, adapted to serum-free suspension culture), without biological pre-enrichment, has been demonstrated. The novel device has been compared to manual silica spin columns, which remain the gold standard in most laboratories. These columns failed to yield the same limit of detection and reproducible results without separating mammalian cells from contaminants. Co-culture experiments have shown that the novel method allows detection of Mollicutes grown for days in presence of mammalian cells, despite the fact that these microorganisms can adhere to eukaryotic cells or invade them. The co-culture data also suggest that the novel sample preparation device might improve the live/dead cells discrimination by removing free nucleic acids from the matrix. PMID- 20026365 TI - Isolation of peptide ligands that interact specifically with human glioma cells. AB - Poor prognosis of high grade gliomas coupled with the difficulty of widespread delivery of therapeutic agents prompted the search into new molecular targets. Our aim is to isolate glioma-specific peptide sequences that can be used for targeted delivery of therapeutic drugs and imaging tracer to accurately demarcate tumor volume as a response to therapy. Herein, we describe the isolation and characterization of a glioma-specific peptide sequence, GL1, that interact exclusively with human glioma cells lines and primary glioma cells derived from human biopsy in vitro. Further analysis showed that the receptors for GL1 were located on the external side of the plasma membrane, where the GL1 peptides could bind stably up to a period of 180 min. More importantly, GL1 phages home specifically to human glioma xenograft when administered through tail vein, a phenomenon that was not observed when non-specific phages were used as control. Taken together, our results confirmed that GL1 could represent a novel peptide that target to tumor of glial origins, and could potentially be used as a targeting moiety for the conjugation of therapeutic drugs or diagnostic imaging radiolabels. PMID- 20026364 TI - Preparation and characterization of salmon calcitonin-sodium triphosphate ionic complex for oral delivery. AB - Even though salmon calcitonin (sCT) has been known as a potent hypocalcemic agent, only injection or nasal spray products are available on the market. In order to develop oral delivery system of the agent, a novel sCT-sodium tripolyphosphate (STPP) ionic complex was fabricated and also characterized. For the optimization of the ionic complexation, the effect of incubation time and molar ratio between sCT and STPP was evaluated. Particle size of the ionic complex in aqueous media, SEM images, DSC, FT-IR, in vitro release test, stability within the simulated intestinal fluid, and hypocalcemic effect were evaluated. The optimal molar complexation ratio of sCT to STPP was ranged from 1:5 to 1:10 and the complexation efficiency was about 95%. The SEM image has shown that the freeze dried ionic complex has rough morphology in their surface and the particle size in PBS (pH 7.4) was about 220nm. The DSC and FT-IR results provided evidences for ionic interaction between -NH(2) groups and -P horizontal lineO groups of sCT and STPP, respectively. The sCT ionic complex has shown sustained sCT releasing characteristics for 3weeks. The sCT-STPP ionic complex was protective to enzymatic attack and in vivo animal data revealed that the present ionic complex would show continuous hypocalcemic effect. Conclusively, the present sCT-STPP ionic complex formulation thought to be a novel oral delivery candidate for the treatment of osteoporosis. PMID- 20026366 TI - Short-term angiotensin-1 receptor antagonism in type 2 diabetic Goto-Kakizaki rats normalizes endothelin-1-induced mesenteric artery contraction. AB - Endothelin (ET)-1 and angiotensin II (Ang II) are likely candidates for a key role in diabetic vascular complications. We demonstrated previously that an enhanced ET-1-induced contraction is present in mesenteric arteries from Goto Kakizaki (GK) rats at the chronic stage of type 2 diabetes. Here, we investigated whether short-term treatment of such rats with losartan, an angiotensin type 1 receptor antagonist, might normalize the ET-1-induced contraction. In mesenteric arteries from GK rats at the chronic stage (34-38 weeks) (vs. those from age matched control Wistar rats): (1) the ET-1-induced contraction was enhanced, (2) the levels of ET-1 and Ang II were increased, (3) ET-1-stimulated ERK2 phosphorylation was increased, and (4) the ACh-induced endothelium-dependent relaxation was reduced. Mesenteric arteries isolated from such GK rats following treatment with losartan (25mg/kg/day for 2 weeks) exhibited reduced ET-1- and Ang II-induced contractions, suppressed ET-1-stimulated ERK phosphorylation, and increased ACh-induced relaxation, while the rats exhibited normalized plasma NO metabolism and their mesenteric arteries exhibited increased basal NO formation. However, such losartan treatment did not alter the increased levels of ET-1 and Ang II seen in GK mesenteric arteries. Our data suggest that within the timescale studied here, losartan normalizes ET-1-induced mesenteric artery contraction through a suppression of ERK activities and/or by normalizing endothelial function. PMID- 20026367 TI - Effect of hydrogen sulfide on intracellular calcium homeostasis in neuronal cells. AB - Hydrogen sulfide (H(2)S) is now known as a new biological mediator. In the present study, the effects of H(2)S on intracellular calcium ([Ca(2+)](i)) in neuronal SH-SY5Y cells was investigated. In SH-SY5Y neuronal cells, NaHS, a H(2)S donor, concentration-dependently increased [Ca(2+)](i). The H(2)S-induced Ca(2+) elevation was significantly attenuated by EGTA-treated calcium-free Krebs' solution. This elevation was also reduced by antagonists of L-type (verapamil and nifedipine), T-type (mibefradil) calcium channels and N-methyl-d-aspartate receptor (MK-801, AP-5 and ifenprodil). A 90% reduction in H(2)S-induced [Ca(2+)](i) elevation was found in cells pretreated with combination of all three kinds of inhibitors. Depletion of intracellular Ca(2+) store with thapsigargin or cyclopiazonic acid or blockade of ryanodine receptor with ruthenium red significantly attenuated the effect of H(2)S on [Ca(2+)](i). Inhibition of protein kinase A (PKA), phospholipase C (PLC) and protein kinase C (PKC) suppressed the H(2)S-elevated [Ca(2+)](i), suggesting that H(2)S may regulate [Ca(2+)](i) via both PKA and PLC/PKC pathways. In conclusion, it was found in this study that H(2)S increased [Ca(2+)](i) in SH-SY5Y neuronal cells by increasing Ca(2+) influx via plasma membrane and in turn releasing calcium from intracellular calcium store. The findings in the present study provide the direct evidence that H(2)S may serve as a neuromodulator. PMID- 20026368 TI - Rat brain docosahexaenoic acid metabolism is not altered by a 6-day intracerebral ventricular infusion of bacterial lipopolysaccharide. AB - In a rat model of neuroinflammation, produced by a 6-day intracerebral ventricular infusion of bacterial lipopolysaccharide (LPS), we reported that the brain concentrations of non-esterified brain arachidonic acid (AA, 20:4 n-6) and its eicosanoid products PGE(2) and PGD(2) were increased, as were AA turnover rates in certain brain phospholipids and the activity of AA-selective cytosolic phospholipase A(2) (cPLA(2)). The activity of Ca(2+)-independent iPLA(2), which is thought to be selective for the release of docosahexaenoic acid (DHA, 22:6 n 3) from membrane phospholipid, was unchanged. In the present study, we measured parameters of brain DHA metabolism in comparable artificial cerebrospinal fluid (control) and LPS-infused rats. In contrast to the reported changes in markers of AA metabolism, the brain non-esterified DHA concentration and DHA turnover rates in individual phospholipids were not significantly altered by LPS infusion. The formation rates of AA-CoA and DHA-CoA in a microsomal brain fraction were also unaltered by the LPS infusion. These observations indicate that LPS-treatment upregulates markers of brain AA but not DHA metabolism. All of which are consistent with other evidence that suggest different sets of enzymes regulate AA and DHA recycling within brain phospholipids and that only selective increases in brain AA metabolism occur following a 6-day LPS infusion. PMID- 20026369 TI - Measuring behavior in mice with chronic stress depression paradigm. AB - Many studies with chronic stress, a common depression paradigm, lead to inconsistent behavioral results. We are introducing a new model of stress-induced anhedonia, which provides more reproducible induction and behavioral measuring of depressive-like phenotype in mice. First, a 4-week stress procedure induces anhedonia, defined by decreased sucrose preference, in the majority of but not all C57BL/6 mice. The remaining 30-50% non-anhedonic animals are used as an internal control for stress effects that are unrelated to anhedonia. Next, a modified sucrose test enables the detection of inter-individual differences in mice. Moreover, testing under dimmed lighting precludes behavioral artifacts caused by hyperlocomotion, a major confounding factor in stressed mice. Finally, moderation of the stress load increases the reproducibility of anhedonia induction, which otherwise is difficult to provide because of inter-batch variability in laboratory mice. We believe that our new mouse model overcomes some major difficulties in measuring behavior with chronic stress depression models. PMID- 20026370 TI - Lorazepam and diazepam for relieving catatonic features precipitated by initial hemodialysis in a uremic patient: a case report. PMID- 20026371 TI - Antidepressant-like action of the ethanolic extract from Tabebuia avellanedae in mice: evidence for the involvement of the monoaminergic system. AB - The antidepressant-like effect of the ethanolic extract obtained from barks of Tabebuia avellanedae, a plant widely employed in folk medicine, was investigated in two predictive models of depression: forced swimming test (FST) and tail suspension test (TST) in mice. Additionally, the mechanisms involved in this antidepressant-like action and the effects of the association of the extract with the antidepressants fluoxetine, desipramine and bupropion in the TST were investigated. The extract from T. avellanedae produced an antidepressant-like effect, in the FST (100 mg/kg, p.o.) and in the TST (10-300 mg/kg, p.o.), without accompanying changes in ambulation when assessed in the open-field test. The anti immobility effect of the extract (30 mg/kg, p.o.) in the TST was prevented by pre treatment of mice with ketanserin (5 mg/kg, i.p., a preferential 5-HT(2A) receptor antagonist), prazosin (1 mg/kg, i.p., an alpha(1)-adrenoceptor antagonist), yohimbine (1 mg/kg, i.p., an alpha(2)-adrenoceptor antagonist), propranolol (2 mg/kg, i.p., a beta-adrenoceptor antagonist), sulpiride (50 mg/kg, i.p., a dopamine D(2) receptor antagonist) and SCH23390 (0.05 mg/kg, s.c., a dopamine D(1) receptor antagonist). The combined administration of a subeffective dose of WAY100635 (0.1 mg/kg, s.c., a selective 5-HT(1A) receptor antagonist) and a subeffective dose of the extract (1 mg/kg, p.o.) produced a significant reduction in the immobility time in the TST. In addition, the combination of fluoxetine (1 mg/kg, p.o.), desipramine (0.1 mg/kg, p.o.), or bupropion (1 mg/kg, p.o.) with a subeffective dose of the extract (1 mg/kg, p.o.) produced a synergistic antidepressant-like effect in the TST, without causing hyperlocomotion in the open-field test. It may be concluded that the extract from T. avellanedae produces an antidepressant-like effect in the FST and in the TST that is dependent on the monoaminergic system. Taken together, our results suggest that T. avellanedae deserves further investigation as a putative alternative therapeutic tool that could help the conventional pharmacotherapy of depression. PMID- 20026373 TI - Naturally occurring phenolic acids inhibit 12-O-tetradecanoylphorbol-13-acetate induced NF-kappaB, iNOS and COX-2 activation in mouse epidermis. AB - The aim of this study was to investigate the effects of naturally occurring protocatechuic, chlorogenic and tannic acids on the skin tumor promoter, 12-O tetradecanoylphorbol-13-acetate (TPA), induced NF-kappaB in mouse epidermis. The topical application of these phenolics 15min prior to TPA resulted in a significant decrease in the NF-kappaB activation which was measured in terms of p65-DNA binding. Tannic acid was the most potent inhibitor of the TPA-stimulated p65-DNA binding, while chlorogenic acid was the least effective compound. Tannic acid also reduced the most the NF-kappaB p65 subunit translocation from cytosol to the nucleus and enhanced the retention of IkappaBalpha in the cytosol. Although protocatechuic acid decreased p65-DNA binding, it did not affect TPA stimulated degradation of IkappaBalpha. All the tested compounds inhibited the IkappaBalpha kinase (IKK) activity in mouse epidermis. Tannic acid was the most potent inhibitor and protocatechuic acid the weakest. Tannic and chlorogenic acids reduced the TPA-induced C-L activity of proteasome 20S to a similar extent. The blockade of upstream kinase IKK signaling by tannic acid, but also by protocatechuic acid, inhibited the enzyme level and the activity of COX-2. Protocatechuic acid also diminished the level and activity of TPA-induced iNOS, which might be related to its weak effect on IkappaBalpha degradation. Our earlier studies demonstrated that these compounds, particularly tannic acid, reduced the formation of the polycyclic aromatic hydrocarbon-DNA adducts in vitro and in vivo in mouse epidermis. The results of our present study indicate that the compounds which reduce the formation of electrophilic PAH metabolites may also diminish NF-kappaB activation. Thus, the phenolic acids, particularly tannic acid, by affecting the key events of initiation and promotion stage of carcinogenesis, have become of great interest for the prevention of cancer. PMID- 20026372 TI - Casiopeina IIgly-induced oxidative stress and mitochondrial dysfunction in human lung cancer A549 and H157 cells. AB - Casiopeinas are a series of mixed chelate copper complexes that are being evaluated as anticancer agents. Their effects in the cell include oxidative damage and mitochondrial dysfunction, yet the molecular mechanisms leading to such effects remain unclear. We tested whether [Cu(4,7-dimethyl phenanthroline)(glycinate)]NO(3) (Casiopeina IIgly or Cas IIgly) could alter cellular glutathione (GSH) levels by redox cycling with GSH to generate ROS and cellular oxidative stress. Cas IIgly induced a dramatic drop in intracellular levels of GSH in human lung cancer H157 and A549 cells, and is able to use GSH as source of electrons to catalyze the Fenton reaction. In both cell lines, the toxicity of Cas IIgly (2.5-5 microM) was potentiated by the GSH synthesis inhibitor l-buthionine sulfoximine (BSO) and diminished by the catalytic antioxidant manganese(III) meso-tetrakis(N,N'-diethylimidazolium-2-yl)porphyrin (MnTDE-1,3-IP(5+)), thus supporting an important role for oxidative stress. Cas IIgly also caused an over-production of reactive oxygen species (ROS) in the mitochondria and a depolarization of the mitochondrial membrane. Moreover, Cas IIgly produced mitochondrial DNA damage that resulted in an imbalance of the expression of the apoproteins of the mitochondrial respiratory chain, which also can contribute to increased ROS production. These results suggest that Cas IIgly initiates multiple possible sources of ROS over-production leading to mitochondrial dysfunction and cell death. PMID- 20026374 TI - Chikungunya fever: CNS infection and pathologies of a re-emerging arbovirus. AB - Chikungunya virus (CHIKV) is transmitted by Aedes mosquitoes and causes an acute symptomatic illness with fever, skin rash, and incapacitating arthralgia, which can evolve into chronic rheumatoid arthritis in elderly patients. This is a tropical disease originally described in central/east Africa in the 1960s, but its 2004 re-emergence in Africa and rapid spread in lands in and around the Indian Ocean (Reunion island, India, Malaysia) as well as Europe (Italy) led to almost 6 million cases worldwide. The risk of importation and spreading diseases with long-term sequelae is even greater today given the global distribution of the vectors (including in the Americas), increased tourism and the apparent capacity of CHIKV to produce high levels of viremia (10(9)-10(12) virus/ml of blood) and new mutants. CHIKV-associated neuropathology was described early in the 1960s, but it is the unprecedented incidence rate in Indian Ocean areas with efficient clinical facilities that allowed a better description of cases with severe encephalitis, meningoencephalitis, peripheral neuropathies and deaths among newborns (mother-to-child infection), infants and elderly patients. Death rates following CHIKV infection were estimated at 1:1000 cases in la Reunion's outbreak. These clinical observations have been corroborated by experimental infection in several mouse models, leading to CNS pathologies. We further describe in this review the capacity of CHIKV to infect neurons and glial cells, delineate the fundamental innate (intrinsic) immune defence mechanisms to protect from infection and argue about the possible mechanisms involved in the encephalopathy. PMID- 20026375 TI - Electrophysiological evidence of interaction between contextual expectation and semantic integration during the processing of collocations. AB - Despite the potentially infinite creativity of language, many words are patterned in ordered strings called collocations. Final words of these clusters are highly predictable; in addition, their overall meaning can vary on the literality dimension, ranging from (figurative) idiomatic strings to literal strings. These structures thus offer a natural linguistic scenario to contrast ERP correlates of contextual expectation and semantic integration processes during comprehension. In this study, expected endings elicited a positive peak around 300ms compared to less expected synonyms, suggesting that the earlier recognition of the string leads to the specific pre-activation of the lexical items that conclude the expression. On the other hand, meaning variations of these fixed strings (either a literal or a figurative whole meaning) affected ERPs only around 400ms, i.e. in the frontal portion of the N400. These findings are discussed within a more general cognitive framework as outlined in Kok's (2001) dual categorization model. PMID- 20026376 TI - Maternal trait anxiety, emotional distress, and salivary cortisol in pregnancy. AB - Animal models suggest that stress-induced hormonal changes in the mother during pregnancy lead to enduring changes in the fetus and empirical links between prenatal maternal stress and negative child development have been discerned repeatedly in human studies. But the role of heritable personality traits has received little attention in the latter work. The goal of the current study was to investigate the relationship between maternal personality, psychological measures of maternal distress and maternal salivary cortisol during pregnancy. Maternal reports of personality (16 PF) and stress-related psychological measures (depression, pregnancy-related anxiety, perceived stress, negative life events) as well as salivary cortisol samples of 66 healthy pregnant women were collected in early and late pregnancy. Maternal trait anxiety proved related to all stress related psychological measures and high anxiety predicted low baseline cortisol awakening levels in early pregnancy. Maternal trait anxiety is related to both psychological and biological stress measures during pregnancy. PMID- 20026377 TI - CD154 expression triggered by purine analogues in vitro: Correlation with treatment response and autoimmune events in chronic lymphocytic leukemia. AB - OBJECTIVE: Despite a fludarabine-based treatment is the first choice of therapy in chronic lymphocytic leukemia (CLL), not all patients achieve a partial or complete response and some of them develop autoimmune manifestations. The aim of this study was to evaluate the influence of CD154 on these adverse effects because CD154 is involved in both B-cell survival and autoimmunity. MATERIALS AND METHODS: Peripheral blood mononuclear cells (PBMC) from 36 patients with CLL were cultured in vitro with fludarabine or 2-chlorodeoxyadenosine for 24, 48, and 72 hours. RESULTS: Seven patients (19.4%) presented CD154 expression in PBMC cultured with purine analogues in vitro for 24 and/or 48 hours, while no expression was found when cultured in media alone. These seven patients showed a decreased apoptotic rate in vitro after purine analogues compared with those patients who did not express CD154 (p = 0.01 for fludarabine; p < 0.001 for 2 chlorodeoxyadenosine). CD154 expression was found to have prognostic value for response to fludarabine in vivo and was associated with the development of autoimmune manifestations (odds ratio = 25; 95% confidence interval = 3.5-166.7; p < 0.001). CONCLUSION: Our preliminary results suggest that CD154 expression in CLL patients, which may be induced by purine analogues, is associated with resistance to fludarabine and with development of autoimmune manifestations. PMID- 20026378 TI - Effects of oral creatine and resistance training on serum myostatin and GASP-1. AB - Myostatin is a catabolic regulator of skeletal muscle mass. The purpose of this study was to determine the effect of resistance training for 8 weeks in conjunction with creatine supplementation on muscle strength, lean body mass, and serum levels of myostatin and growth and differentiation factor-associated serum protein-1 (GASP-1). In a double-blinded design 27 healthy male subjects (23.42+/ 2.2 years) were assigned to control (CON), resistance training+placebo (RT+PL) and resistance training+creatine supplementation (RT+CR) groups. The protocol consisted of 3 days per week of training for 8 weeks, each session including three sets of 8-10 repetitions at 60-70% of 1 RM for whole-body exercise. Blood sampling, muscular strength testing and body composition analysis (full body DEXA) were performed at 0, 4th and 8th weeks. Myostatin and GASP-1 was measured. Resistance training caused significant decrease in serum levels of myostatin and increase in that of GASP-1. Creatine supplementation in conjunction with resistance training lead to greater decreases in serum myostatin (p<0.05), but had not additional effect on GASP-1 (p>0.05). The effects of resistance training on serum levels of myostatin and GASP-1, may explain the increased muscle mass that is amplified by creatine supplementation. PMID- 20026379 TI - Metabolic signals in human puberty: effects of over and undernutrition. AB - Puberty in mammals is associated with important physical and psychological changes due to the increase in sex steroids and growth hormone (GH). Indeed, an increase in growth velocity and the attainment of sexual maturity for future reproductive function are the hallmark changes during this stage of life. Both growth and reproduction consume high levels of energy, requiring suitable energy stores to face these physiological functions. During the last two decades our knowledge concerning how peptides produced in the digestive tract (in charge of energy intake) and in adipose tissue (in charge of energy storage) provide information regarding metabolic status to the central nervous system (CNS) has increased dramatically. Moreover, these peptides have been shown to play an important role in modulating the gonadotropic axis with their absence or an imbalance in their secretion being able to disturb pubertal onset or progression. In this article we will review the current knowledge concerning the role played by leptin, the key adipokine in energy homeostasis, and ghrelin, the only orexigenic and growth-promoting peptide produced by the digestive tract, on sexual development. The normal evolutionary pattern of these peripherally produced metabolic signals throughout human puberty will be summarized. The effect of two opposite situations of chronic malnutrition, obesity and anorexia, on these signals and how they influence the course of puberty will also be discussed. Finally, we will briefly mention other peptides derived from the digestive tract (such as PYY) that may be involved in the regulatory link between energy homeostasis and sexual development. PMID- 20026380 TI - Electrophysiological evidence for effects of color knowledge in object recognition. AB - Knowledge about the typical colors associated with familiar everyday objects (i.e., strawberries are red) is well-known to be represented in the conceptual semantic system. Evidence that such knowledge may also play a role in early perceptual processes for object recognition is scant. In the present ERP study, participants viewed a list of object pictures and detected infrequent stimulus repetitions. Results show that shortly after stimulus onset, ERP components indexing early perceptual processes, including N1, P2, and N2, differentiated between objects in their appropriate or congruent color from these objects in an inappropriate or incongruent color. Such congruence effect also occurred in N3 associated with semantic processing of pictures but not in N4 for domain-general semantic processing. Our results demonstrate a clear effect of color knowledge in early object recognition stages and support the following proposal-color as a surface property is stored in a multiple-memory system where pre-semantic perceptual and semantic conceptual representations interact during object recognition. PMID- 20026381 TI - Gamma-secretase inhibitor (GSI1) attenuates morphological cerebral vasospasm in 24h after experimental subarachnoid hemorrhage in rats. AB - Notch signaling plays an important role in the arteriogenesis. We hypothesized that the Notch inhibitor--gamma-secretase inhibitor (GSI1) exerted its effects on the vasospasm via regulation of NF-kappaB and MMP-9. In this study, 160 male Sprague-Dawley (SD) rats were randomly assigned into four groups: Sham, subarachnoid hemorrhage (SAH), SAH treated with dimethyl sulfoxide (DMSO) and SAH treated with GSI1. After 24h SAH, the mortality, neurological scores, blood-brain barrier permeability and brain water content were examined. The mRNA and protein level of Notch1, the expression and activity of NF-kappaB and MMP-9 were evaluated. Severe morphological vasospasm in the basilar artery was observed in SAH and DMSO treated rats. GSI1 significantly effected on neurological deficits, but not on mortality; significantly reduced morphological vasospasm, blood-brain barrier permeability, brain water content; significantly decreased the protein level of Notch1, NF-kappaB p50 and MMP-9, as well as the DNA-binding activity of NF-kappaB (EMSA) and the activity of MMP-9 (Zymography). These findings suggest that GSI1 plays a critical role in the attenuation of acute cerebral vasospasm, which may provide a novel therapeutic target for cerebral vasospasm after SAH insult. PMID- 20026382 TI - Intrathecal huperzine A increases thermal escape latency and decreases flinching behavior in the formalin test in rats. AB - Huperzine A (HupA) is an alkaloid isolated from the Chinese club moss Huperzia serrata and has been used for improving memory, cognitive and behavioral function in patients with Alzheimer's disease in China. It has NMDA antagonist and anticholinesterase activity and has shown anticonvulsant and antinociceptive effects in preliminary studies when administered intraperitoneally to mice. To better characterize the antinociceptive effects of HupA at the spinal level, Holtzman rats were implanted with intrathecal catheters to measure thermal escape latency using Hargreaves thermal escape testing system and flinching behavior using the formalin test. Intrathecal (IT) administration of HupA showed a dose dependent increase in thermal escape latency with an ED50 of 0.57 microg. Atropine reversed the increase in thermal escape latency produced by 10 microg HupA, indicating an antinociceptive mechanism through muscarinic cholinergic receptors. The formalin test showed that HupA decreased flinching behavior in a dose-dependent manner. Atropine also reversed the decrease in flinching behavior caused by 10 microg HupA. A dose-dependent increase of side effects including scratching, biting, and chewing tails was observed, although antinociceptive effects were observed in doses that did not produce any adverse effects. PMID- 20026383 TI - Influence of sensory inputs and motor demands on the control of the centre of mass velocity during gait initiation in humans. AB - Human gait requires the simultaneous generation of goal-directed continuous movement (locomotion) and the maintenance of balance (postural control). In adults, the centre of mass (CoM) oscillates in the vertical plane while walking. During the single support phase of gait initiation, its vertical (vCoM) velocity increases as the CoM falls and is actively reversed prior to foot-contact. In this study we investigated whether this active control, which is thought to reflect balance control during gait initiation, is controlled by visual and somatosensory inputs (Experiment 1) and whether it is modified by a change in motor demands, two steps versus one step (Experiment 2). In all healthy adults, the vCoM velocity was braked, or controlled, by contraction of the soleus muscle of the stance leg. The elimination of visual input alone had no effect on braking, although its amplitude decreased when somatosensory inputs were disrupted (-47%), and further decreased when both visual and somatosensory inputs were disrupted (-83%). When subjects performed only one step, with no trailing of the stance foot, the vCoM velocity braking also decreased (-42%). These results suggest that active braking of the CoM fall during the transition to double support, an indicator of balance control, is influenced by both multisensory integration and the demands of the current motor program. The neural structures involved in this mechanism remain to be elucidated. PMID- 20026384 TI - Anticonvulsant doses of ganaxolone do not compromise motor performance in immature rats. AB - Neuroactive steroids that function as positive modulators of GABA-A receptors are potential anticonvulsant drugs. We previously demonstrated that ganaxolone is effective against pentetrazol-induced motor seizures in immature rats. In the present study, we examined the effects of ganaxolone in another model, cortical epileptic afterdischarges (ADs). The possible side effects of ganaxolone were studied in rats 12, 18, and 25 days of age following the implantation of epidural electrodes. Low-frequency stimulation of the sensorimotor cortical area elicited ADs characterized by a spike-and-wave rhythm and clonic seizures. Ganaxolone (5, 10, 20, or 40 mg/kg) was administered intraperitoneally after the first AD and stimulation was repeated five more times. The highest dose of ganaxolone (40 mg/kg) suppressed progressive prolongation of ADs in 25-day-old rats and postponed it in 12-day-old rats. No significant effect was observed in 18-day-old animals. Movements during stimulation and clonic seizures accompanying ADs were not affected by ganaxolone. Ganaxolone at doses of 20 and 40 mg/kg had no significant effect on motor function, such as surface righting, negative geotaxis, wire mesh ascending, and bar holding. After administration of 40 mg/kg ganaxolone to 18- and 25-day-old rats, spontaneous locomotion in the open field tended to decrease. Doses of ganaxolone with a moderate anticonvulsant effect in the present model did not seriously compromise motor performance. PMID- 20026385 TI - Selenium attenuates A beta production and A beta-induced neuronal death. AB - The objective of the present study was to examine the role of selenium in the metabolism of A beta and in A beta-induced neuronal death. Selenium treatment significantly reduced A beta 40, A beta 42, and sAPP beta production by reducing A beta producing beta-secretase and gamma-secretase activities. The lipid peroxidation product 4-Hydroxynonenal (HNE)-induced transcription of beta secretase (BACE1) was blocked by selenium. Finally, our data show that selenium protects against HNE and A beta-mediated toxicity in primary cultured neurons. The present study suggests that selenium may be able to salvage the neuronal degeneration of Alzheimer's disease, thereby limiting beta-amyloid production and neuronal death. PMID- 20026387 TI - Sound exposure accelerates reflex emergence and development in young rats. AB - Early sensory experience affects brain development. In rats, most somatic reflexes are not expressed at birth but may take as long as 2 weeks to emerge. Whether sensory enrichment during this early period affects reflex maturation remains unknown. Here, we exposed rat pups to a pure tone (4kHz, 65dB SPL, 8h/day) with their nursing mother during the first 3 postnatal weeks and measured the times when reflexes appeared on the basis of video recordings. Sound exposure accelerated by about 15% the appearance of all reflexes assessed (righting, cliff avoidance, vibrissa placing, negative geotaxis and auditory startle, p<0.001). In addition, sound exposure accelerated the appearance of developmental characteristics: incisor eruption, ear unfolding and eye opening. These changes occurred concomitantly with an increase in pups' body and brain weights, together with a dramatic increase in fluid intake of the nursing mother. These findings are the first evidence that early sound exposure, even before opening of ear canals, accelerates reflex development. We speculate that the observed changes could involve the nursing mother. PMID- 20026388 TI - Lactoferrin-modified nanoparticles could mediate efficient gene delivery to the brain in vivo. AB - Lactoferrin (Lf)-modified nanoparticles (NPs) have been demonstrated to mediate efficient expression of exogenous genes in the brain via intravenous administration. The brain-targeting properties of Lf-modified NPs were investigated in this study. In vivo imaging results showed that the accumulation of Lf-modified NPs was higher in the brain but lower in the other organs than that of unmodified counterparts. The results of analytical transmission electron microscopy showed that some Lf-modified NPs crossed the blood-brain barrier (BBB) and reached the neural tissues, while some remained within the BBB. Similar results were observed in the distribution of exogenous gene products. All the results demonstrated the successful delivery of Lf-modified NPs into the brain. Lf-modified NPs could be exploited as potential brain-targeting delivery systems for exogenous genes, especially for those encoding secretive proteins. PMID- 20026386 TI - The ferritin superfamily: Supramolecular templates for materials synthesis. AB - Members of the ferritin superfamily are multi-subunit cage-like proteins with a hollow interior cavity. These proteins possess three distinct surfaces, i.e. interior and exterior surfaces of the cages and interface between subunits. The interior cavity provides a unique reaction environment in which the interior reaction is separated from the external environment. In biology the cavity is utilized for sequestration of irons and biomineralization as a mechanism to render Fe inert and sequester it from the external environment. Material scientists have been inspired by this system and exploited a range of ferritin superfamily proteins as supramolecular templates to encapsulate nanoparticles and/or as well-defined building blocks for fabrication of higher order assembly. Besides the interior cavity, the exterior surface of the protein cages can be modified without altering the interior characteristics. This allows us to deliver the protein cages to a targeted tissue in vivo or to achieve controlled assembly on a solid substrate to fabricate higher order structures. Furthermore, the interface between subunits is utilized for manipulating chimeric self-assembly of the protein cages and in the generation of symmetry-broken Janus particles. Utilizing these ideas, the ferritin superfamily has been exploited for development of a broad range of materials with applications from biomedicine to electronics. PMID- 20026389 TI - Presence of mRNA of muscle nicotinic acetylcholine receptor subunits and an epsilon-subunit splice variant in the mouse brain. AB - Transcripts encoding for alpha1, beta1, delta, gamma and epsilon (and its splice variant epsilon(s)) subunits of the muscle-type nicotinic acetylcholine receptor (nAChR) were assessed using reverse transcription followed by polymerase chain reaction (RT-PCR) assays, with RNA extracted from the mouse skeletal muscle (diaphragm) and brain regions (cortex, hippocampus and cerebellum). The presence of alpha1, beta1, delta, gamma, epsilon and epsilon(s) transcripts was confirmed in the diaphragm muscle, used as positive control. mRNAs coding for muscle alpha1, beta1, delta, epsilon, epsilon(s), but not gamma subunits, were detected in adult mouse brain regions. An epsilon-subunit sequence variant, named epsilon(t), was also detected in all brain regions examined, but not in skeletal muscle. This new epsilon-subunit splice variant lacks a 115 bp cassette corresponding to exon 8 in the first intracellular transmembrane domain of the subunit, leading to a truncated protein. The data provide evidence for the presence of muscle-type nAChR subunits in the mouse central nervous system. PMID- 20026390 TI - Preparation of icariside II from icariin by enzymatic hydrolysis method. AB - It has been reported that icariin and icariside II, two flavonoid glycosides coming from herba epimedii, which have a closely structural relationship, show some pharmacological effects such as preventing osteoporosis, cancer and depression. The content of natural icariside II is very low in herba epimedii, but it is the main component in vivo after the administration of herba epimedii. More icariside II can be obtained from icariin by enzymatic hydrolysis method than by traditional isolation method. This study focuses on finding a simple and feasible method to prepare icariside II from icariin by enzymatic hydrolysis, so as to meet the request for further pharmacologic actions study. Icariin was obtained successively with 90% ethanol extraction, isolation on macroporous resin and purification on silica gel chromatography. Enzymatic hydrolysis conditions were tested for the bioconversion of icariin into icariside II by orthogonal array design. The structures of isolated icariin and produced icariside II were identified by UV, IR, ESIMS, (1)H NMR, (13)C NMR, and DEPT spectroscope. Enzymatic hydrolysis experiment showed that icariin could be transformed into icariside II with the action of beta-glucosidase and the optimum reaction conditions were determined as follows: 50 degrees C, 0.2 M disodium hydrogen phosphate and citric acid buffer system (pH6.0), the ratio of icariin/enzyme is 1:1 and reaction time 5 h. By using this enzymatic condition, 95.5 mg icariside II (with the purity of 99.1%) was obtained eventually by transforming 200 mg icariin. PMID- 20026391 TI - Introduction. PMID- 20026392 TI - Gender and species differences in triadimefon metabolism by rodent hepatic microsomes. AB - Understanding the potential differences in metabolic capacity and kinetics between various common laboratory species as well as between genders is an important facet of chemical risk assessment that is often overlooked, particularly for chemicals which undergo non-cytochrome P450 mediated metabolism. The use of physiologically based pharmacokinetic (PBPK) models to better describe chemical exposure is made more powerful by incorporation of high quality in vitro kinetic data. To this end, metabolism of the conazole fungicide triadimefon was studied in hepatic microsomes of both genders of SD rats and CD-1 mice. Triadimefon depletion and triadimenol formation were measured in each type of microsomes. Michaelis-Menten regressions were applied to metabolic data and V(MAX) and the Michaelis constant (K(M)) values calculated. Male SD rats metabolized triadimefon more rapidly than female SD rats or either gender of CD-1 mouse. K(M) values were in the micromolar range, indicating the possibility of competitive inhibition with endogenous substrates. Intrinsic clearances derived from kinetic parameters indicate that triadimefon metabolism is blood-flow limited in all organisms studied with the possible exception of female rat. The in vitro half-life method was investigated as a less resource intensive method for the derivation of intrinsic clearance, and was found to be useful as a complement to the traditional Michaelis-Menten approach. PMID- 20026393 TI - Beta-glucuronidase activity is a sensitive biomarker to assess low-level organophosphorus insecticide exposure. AB - Acetylcholinesterase and butyrylcholinesterase (BChE) activities in blood are widely used as the biomarkers for organophosphorus insecticide (OP) exposure. In the present study, we conducted a cross-sectional study to evaluate plasma beta glucuronidase (BG), a sensitive biomarker candidate for OP exposure, BChE activities and urinary dialkyl phosphates (DAPs), OP metabolites. We assessed the relationship between these biomarker levels in the following groups: 32 controls (control), 21 pest control operators and their co-workers who had not sprayed OPs within 3 days prior to sample collection (PCO1), and 21 pest control operators who sprayed OPs within those 3 days (PCO2). Logarithmically transformed age adjusted means of DAPs were 3.88, 5.62 and 6.45 nmol/g creatinine for control, PCO1 and PCO2, respectively (P<0.001 for difference, P<0.001 for trend). Logarithmically transformed age-adjusted means of BG were 1.40, 1.52 and 1.85 micromol/L/h for control, PCO1 and PCO2, respectively. BG activity, but not BChE, was increased according to their OP exposure level (P=0.038 for difference, P=0.026 for trend). It was concluded that plasma BG activity is more sensitive biomarker as well as urinary OP metabolites than BChE for low-level exposure in humans. PMID- 20026394 TI - Prospective follow-up studies found no chromosomal mutagenicity of methylphenidate therapy in ADHD affected children. AB - Five to ten percent of all children suffer from attention-deficit/hyperactivity disorder (ADHD), which is often treated with the central nervous stimulant methylphenidate (MPH). In 2005 controversy arose due to a report of enhanced cytogenetic effects in 12 children after 3 months of MPH treatment. Since then, several prospective studies have been performed and published, which are summarized here. A table comparing the micronucleus frequencies, a marker investigated in all of these studies, is presented. An induction of cytogenetic effects by MPH was only reported in one, the 2005 study by El-Zein et al., while all other studies, with now altogether 110 MPH-exposed individuals, showed no elevation. To address the question of long-term use of MPH, we published the data of 30 chronically treated children and also saw no difference compared to untreated children. Here, we report as new follow-up data that an additional 12 months time point in a small group of 12 children who had begun MPH therapy within our published study also did not reveal elevated cytogenetic damage. Furthermore, a previously unpublished analysis of urinary 8-oxo-7,8-dihydro-2' deoxyguanosine (8-oxodG; a non-invasive biomarker for DNA-base oxidation and its repair) in 11 children before and after 3 months of MPH exposure yielded no significant difference. Since gene mutations may not necessarily manifest as chromosomal aberrations, micronuclei or SCEs, we discuss the available data from animal models, which also do not reveal a mutagenic potential of MPH. Although the only two available epidemiological studies do not report elevated risk for MPH exposure, the results are not conclusive yet, and further monitoring of exposed populations is suggested. PMID- 20026396 TI - Anti-inflammatory evaluation of Coronopus didymus in the pleurisy and paw oedema models in mice. AB - ETHNOPHARMACOLOGICAL RELEVANCE: Mastrunco (Coronopus didymus--CD) is currently considered as a medicinal specie often used in Brazil, especially in southeast region, for the treatment of several diseases in which pain and inflammation are common. Treatment with the plant can be done by infusion, decoction, or through food. The aim of this study was: to investigate the anti-inflammatory effect of hydroalcoholic extract obtained from the leaves of CD following the traditional procedure. MATERIALS AND METHODS: The anti-inflammatory activity was determined using mouse of pleurisy and paw oedema models, both process being induced by different flogistic agents such as: carrageenan (Cg), bradykinin (BK), histamine (HIS), substance P (SP), dextran (DEX) or prostaglandin E(2) (PGE(2)). We evaluated the effect of CD (200-600 mg/kg) administered by oral route (p.o.) upon leukocytes migration, myeloperoxidase (MPO), and adenosine-deaminase (ADA) activities and nitric oxide (NO) levels. RESULTS: CD (200-600 mg/kg) inhibited the leukocytes by 60.0+/-1.42%, neutrophils by 82.75+/-1.29%, MPO by 42.30+/ 4.23%, and ADA activities by 57.89+/-1.94%, as well as NO levels by 64.28+/-2.15% in Cg induced pleurisy. CD also inhibited total and differential leukocytes in the pleurisy induced by BK (1.30+/-0.11/0.29+/-0.02), HIS (1.20+/-0.09/0.42+/ 0.05) and SP (0.74+/-0.06/0.14+/-0.01). In addition, CD was effective in reducing paw oedema induced by Cg by 72.79+/-1.13%, SP by 68.26.+/-0.78%, BK by 66.66.+/ 0.77%, PGE(2) by 53.346.+/-1.18 and DEX by 65.14+/-2.35%. CONCLUSION: Several mechanisms, including the inhibition of enzymes (MPO and ADA) and mediators (BK, HIS, SP, NO and PGE(2)) release and/or action, appear to account for the anti inflammatory effect of Coronopus didymus. PMID- 20026395 TI - Goniothalamin-induced oxidative stress, DNA damage and apoptosis via caspase-2 independent and Bcl-2 independent pathways in Jurkat T-cells. AB - Goniothalamin (GTN) isolated from Goniothalamus sp. has been demonstrated to induce apoptosis in a variety of cancer cell lines including Jurkat T leukemia cells. However, the mechanism of GTN-induced apoptosis upstream of mitochondria is still poorly defined. In this study, GTN caused a decrease in GSH with an elevation of reactive oxygen species as early as 30 min and DNA damage as assessed by Comet assay. Analysis using topoisomerase II processing of supercoiled pBR 322 DNA showed that GTN caused DNA damage via a topoisomerase II independent pathway suggesting that cellular oxidative stress may contribute to genotoxicity. A 12-fold increase of caspase-2 activity was observed in GTN treated Jurkat cells after 4h treatment and this was confirmed using Western blotting. Although the caspase-2 inhibitor Z-VDVAD-FMK inhibited the proteolytic activity of caspase-2, apoptosis ensued confirming that caspase-2 activity was not crucial for GTN-induced apoptosis. However, GTN-induced apoptosis was completely abrogated by N-acetylcysteine further confirming the role of oxidative stress. Since cytochrome c release was observed as early as 1h without any appreciable change in Bcl-2 protein expression, we further investigated whether overexpression of Bcl-2 confers resistance in GTN-induced cytotoxicity. Using a panel of Jurkat Bcl-2 transfectants, GTN cytotoxicity was not abrogated in these cells. In conclusion, GTN induces DNA damage and oxidative stress resulting in apoptosis which is independent of both caspase-2 and Bcl-2. PMID- 20026397 TI - Protective effect of Calendula officinalis extract against UVB-induced oxidative stress in skin: evaluation of reduced glutathione levels and matrix metalloproteinase secretion. AB - BACKGROUND AND PURPOSE: Calendula officinalis flowers have long been employed time in folk therapy, and more than 35 properties have been attributed to decoctions and tinctures from the flowers. The main uses are as remedies for burns (including sunburns), bruises and cutaneous and internal inflammatory diseases of several origins. The recommended doses are a function both of the type and severity of the condition to be treated and the individual condition of each patient. Therefore, the present study investigated the potential use of Calendula officinalis extract to prevent UV irradiation-induced oxidative stress in skin. METHODS: Firstly, the physico-chemical composition of marigold extract (ME) (hydroalcoholic extract) was assessed and the in vitro antioxidant efficacy was determined using different methodologies. Secondly, the cytotoxicity was evaluated in L929 and HepG2 cells with the MTT assay. Finally, the in vivo protective effect of ME against UVB-induced oxidative stress in the skin of hairless mice was evaluated by determining reduced glutathione (GSH) levels and monitoring the secretion/activity of metalloproteinases. RESULTS AND CONCLUSIONS: The polyphenol, flavonoid, rutin and narcissin contents found in ME were 28.6 mg/g, 18.8 mg/g, 1.6 mg/g and 12.2mg/g, respectively and evaluation of the in vitro antioxidant activity demonstrated a dose-dependent effect of ME against different radicals. Cytoxicity experiments demonstrated that ME was not cytotoxic for L929 and HepG2 cells at concentrations less than or equal to of 15 mg/mL. However, concentrations greater than or equal to 30 mg/mL, toxic effects were observed. Finally, oral treatment of hairless mice with 150 and 300 mg/kg of ME maintained GSH levels close to non-irradiated control mice. In addition, this extract affects the activity/secretion of matrix metalloproteinases 2 and 9 (MMP 2 and -9) stimulated by exposure to UVB irradiation. However, additional studies are required to have a complete understanding of the protective effects of ME for skin. PMID- 20026398 TI - Evaluation of the subchronic toxicity of oral treatment with Chenopodium ambrosioides in mice. AB - AIM OF THE STUDY: The leaves of Chenopodium ambrosioides L. (Chenopodiaceae) have been used by native people to treat many diseases. Recently, we showed that the treatment with small dose (5mg/kg) of hydroalcoholic extract (HE) from Chenopodium ambrosioides' leaves has immunestimulatory effects. The aim of this study was to investigate the subchronic toxicity of the oral treatment with this HE in preclinical assays. MATERIAL AND METHODS: Swiss mice were divided into 4 groups (n=10/group). They received the HE daily at the doses of 5, 50 and 500 mg/kg by gavage during 15 days. The control group received only water. They were observed each hour for 24h and each day for 15 days, when the blood was collected. The serum was used to perform the biochemical analysis. The mice were then killed and the vital and lymphoid organs were collected and evaluated. RESULTS: There was neither death nor alterations in the body weight in the HE treated groups, but there were alterations in the weight of some organs. There was an increase in the lymph node cells number in the highest two doses. The number of cells in the bone marrow was high in the HE-treated groups, but the number of peritoneal cells was smaller in the HE-treated groups when compared to the control. There was no alteration in the AST, but there was a reduction in the albumin levels in the HE500 group and in the triglycerides and VLDL in the highest doses. CONCLUSION: The subchronic treatment with HE induced punctual alterations in the groups treated with the highest doses. However, the HE treatment was not lethal and did not induce toxic alterations using the therapeutic dose, suggesting that it is safe to use this product in the adequate dose. PMID- 20026399 TI - The role of diclofenac sodium in the dimorphic transition in Candida albicans. AB - Diclofenac sodium is a non-steroidal anti-inflammatory drug that inhibits filamentation in Candida albicans. Here we examined the effect of diclofenac sodium on hypha formation in C. albicans. The C. albicans cells were treated with various concentrations of diclofenac sodium (50, 100, 200 and 500microg/ml) and incubated at 37 degrees C for 2h. The characteristics of hypha formation were then assessed microscopically in both liquid and solid media. The results indicated that the effect of diclofenac sodium was dependent on the concentration of this compound, and preincubation with 500microg/ml diclofenac sodium completely inhibited hypha formation in both liquid and solid media. RT-qPCR analysis of RNA extracted from C. albicans indicated that the levels of expression of agglutinin-like sequence 3 (ALS3), RAS1, EFG1 mRNA, which are regulated by the cAMP-EFG1 pathway in C. albicans and three hypha-specific genes (ALS1, ECE1 and HWP1), were decreased in diclofenac sodium treated cells compared to the levels in controls. Our results also demonstrated that diclofenac sodium possesses potent anti yeast-hypha transition activity in vitro and it could be useful in combined therapy with conventional antifungal agents in the management of treatment of Candida albicans infections. PMID- 20026400 TI - Beneficial effects of PTP1B deficiency on brown adipocyte differentiation and protection against apoptosis induced by pro- and anti-inflammatory stimuli. AB - Insulin is an inducer of brown fat adipogenesis through the activation of a signalling network that involves positive/negative modulators. Given the importance of brown adipose tissue (BAT) for basal thermogenic energy expenditure, we investigated the role of PTP1B in the acquisition of terminal differentiated phenotype and in the apoptotic responses of brown adipocytes. Immortalized brown preadipocytes lacking (PTP1B(-/-)) or expressing (PTP1B(+/+)) PTP1B have been generated. PTP1B deficiency accelerated a full program of brown adipogenesis including induction of transcription factors, coactivators, adipogenic markers and signalling molecules. Fully differentiated PTP1B(-/-) brown adipocytes were resistant to tumor necrosis factor (TNFalpha)-induced apoptosis as these cells were protected against caspase-8 activation, FLIP degradation, Bid cleavage and caspase-3 activation compared to wild-type controls. These events were recovered by PTP1B rescue. Survival signalling including phosphorylation of IRS-1 and Akt/PKB and BclxL expression were decreased in TNFalpha-treated PTP1B(-/-) cells but not in the wild-type. Similarly, PTP1B(-/-) brown adipocytes were protected against resveratrol-induced apoptosis. Phosphorylation of Akt/PKB and Foxo1 phosphorylation/acetylation decreased exclusively in resveratrol-treated wild-type cells, leading to nuclear localization of Foxo1 and up-regulation of Bim. Thus, PTP1B inhibition could be of benefit against obesity by counteracting TNFalpha-induced brown fat atrophy, and combined with resveratrol might improve low-grade inflammation. PMID- 20026401 TI - Preparation of sustained release matrix pellets by melt agglomeration in the fluidized bed: influence of formulation variables and modelling of agglomerate growth. AB - The one-step preparation of sustained release matrix pellets, using a melting procedure in a fluidized bed apparatus, was tested in a 2(3) full factorial design of experiments, using microcrystalline wax as lipophilic binder, theophylline as model drug and talc as additional matrix forming agent. The three influence parameters were (A) size of binder particles, (B) fraction of theophylline in solid particles and (C) fraction of microcrystalline wax in formulation. The response variables were agglomerate size and size distribution, dissolution time, agglomerate crush resistance, sphericity, yield and porosity. Nearly spherical pellets comprising a smooth, closed surface could be obtained with the used method, exhibiting the hollow core typical for the immersion and layering mechanism. The reproducibility was very good concerning all responses. The size of agglomerates is proportional to the size of the binder particles, which serve as cores for pellet formation in the molten state in the fluidized bed. Additionally, the agglomerate size is influenced by the volume of the solid particles in relation to the binder particles, with more solid particles leading to larger agglomerates and vice versa. Dissolution times vary in a very wide range, resulting from the interplay between amount of drug in relation to the meltable matrix substance microcrystalline wax and the non-meltable matrix substance talc. The change of binder particle size does not lead to a structural change of the matrix; both dissolution times and porosity are not significantly altered. Agglomerate crush resistance is low due to the hollow core of the pellets. However, it is significantly increased if the volume fraction of microcrystalline wax in the matrix is high, which means that the matrix is mechanically better stabilized. A theoretical model has been established to quantitatively explain agglomerate growth and very good accordance of the full particle size distributions between predicted and actual values could be shown. A low volumetric binder to solids ratio is compensated by a more porous layer. On the basis of this model, in-depth understanding on the mechanism and influence of product properties could be gained; and an a priori estimation of particle size distributions for new formulas can be performed, with densities, formula, and binder particle size distribution as input parameters. PMID- 20026403 TI - Using human neural crest-derived progenitor cells to investigate osteogenesis: an in vitro study. AB - Human tooth contains a distinct population of neural crest-derived progenitor cells (dNC-PCs) which are known to give rise to specialized daughter cells of an osteogenic lineage. We hypothesised that dNC-PCs could develop into neural crest derived bone in a self-propagating and extracorporal culture system. Thus, we examined the three-dimensional structure obtained from osteogenic-stimulated dNC PCs by morphological, biochemical and spectroscopic methods. After the onset of stimulation, cells formed a multilayer with outer cells covering the surface and inner cells secreting a hyaline matrix. With prolonged culture, multilayers contracted and formed a three-dimensional construct which subsequently converted to a calcified mass. Differentiation of progenitor cells was associated with apoptosis. Cell types which survived were smooth muscle actin-positive cells and bone-like cells. The expression of osteoblastic markers and the secretion of a collagenous matrix indicate that the bone cells had acquired their functional phenotype. Furthermore, these cells produced and secreted membrane-bound vesicles into the newly forming matrix. Consequently, an early biomineralized extracellular matrix was found with calcium phosphate deposits being associated with the newly formed collagen matrix framework. The molar calcium-phosphorus ratio of the mineralized collagen indicated that amorphous calcium phosphate was present within this matrix. The data suggest that stimulated cultures of dNC-PCs are able to recapitulate some processes of the early phase of osteogenesis. PMID- 20026402 TI - Characterizing molecular diffusion in the lens capsule. AB - The lens capsule compartmentalizes the cells of the avascular lens from other ocular tissues. Small molecules required for lens cell metabolism, such as glucose, salts, and waste products, freely pass through the capsule. However, the lens capsule is selectively permeable to proteins such as growth hormones and substrate carriers which are required for proper lens growth and development. We used fluorescence recovery after photobleaching (FRAP) to characterize the diffusional behavior of various sized dextrans (3, 10, 40, 150, and 250 kDa) and proteins endogenous to the lens environment (EGF, gammaD-crystallin, BSA, transferrin, ceruloplasmin, and IgG) within the capsules of whole living lenses. We found that proteins had dramatically different diffusion and partition coefficients as well as capsule matrix binding affinities than similar sized dextrans, but they had comparable permeabilities. We also found ionic interactions between proteins and the capsule matrix significantly influence permeability and binding affinity, while hydrophobic interactions had less of an effect. The removal of a single anionic residue from the surface of a protein, gammaD-crystallin [E107A], significantly altered its permeability and matrix binding affinity in the capsule. Our data indicated that permeabilities and binding affinities in the lens capsule varied between individual proteins and cannot be predicted by isoelectric points or molecular size alone. PMID- 20026404 TI - Effect of activation of viral receptors on the gelatinases MMP-2 and MMP-9 in human mesothelial cells. AB - BACKGROUND: Extracellular matrix (ECM) not only provides molecular and spatial information that influence cell proliferation, differentiation and apoptosis but also has the potential to bind and present or release cytokines and cytotactic factors. Synthesis and degradation of extracellular matrix components are balanced by matrix metalloproteinases (MMP) and their inhibitors. In the pericardium as well as in the pleural and peritoneal cavities a multitude of clinically relevant disease states ranging from inflammation to fibrosis and tumor invasion result from altered regulation of MMP activity and are known to be associated with viral disease. METHODS: Therefore, the functional linkage between viral receptors of the innate immune system, the toll-like receptors (TLR), and control of MMP activity was exemplarily analyzed by stimulating human mesothelial cells with poly (I:C) RNA. RESULTS: We hereby show that human mesothelial cells (MC) express TLR3. After stimulation of MC with the cytokines TNF-alpha, IL-1beta and IFN-gamma alone or in combination to simulate a proinflammatory milieu as would occur during immune-mediated inflammatory disease, an upregulation of TLR3 is seen. Furthermore, a selectively TLR3 mediated, time- and dose-dependent upregulation of MMP-9 and TIMP-1 is found, whereas MMP-2 expression is not significantly affected by TLR3 stimulation. CONCLUSIONS: With these results we provide evidence for a mechanism by which infectious agents can mediate processes of the final common path of inflammation as fibrosis via regulation of MMP and TIMP. PMID- 20026405 TI - Leukemia stem cells. AB - Leukemia stem cells (LSC) reside within a hierarchy of malignant hematopoiesis and possess the ability to instigate, maintain and serially propagate leukemia in vivo, while retaining the capacity to differentiate into committed progeny that lack these properties. In most cases, LSC appear to share immunophenotypic characteristics with committed hematopoietic progenitors, however have pathologically enhanced self-renewal, mediated through the activation of certain cellular pathways. The presence of a LSC that solely possesses the ability to initiate and sustain leukemia has implications for the treatment of patients with this disease. In this review, we will discuss these issues as well as some of the recent controversies regarding LSC frequency and alternative theories of leukemogenesis. PMID- 20026406 TI - Alternatively spliced N-terminal exons in tropomyosin isoforms do not act as autonomous targeting signals. AB - Tropomyosin (Tm) polymerises head-to-tail to form a continuous polymer located in the major groove of the actin filament. Multiple Tm isoforms are generated by alternative splicing of four genes, and individual isoforms show specific localisation patterns in many cell types, and can have differing effects on the actin cytoskeleton. Fluorescently-tagged Tm isoforms and mutants were expressed in C2C12 cells to investigate the mechanisms of alternative localisation of high molecular weight (HMW) and low molecular weight (LMW) Tms. Fluorescently-tagged Tm constructs show similar localisation to endogenous Tms as observed by antibodies, with the HMW Tm3 relatively diminished at the periphery of cells compared to LMW isoforms Tm5b or Tm5NM1. Tm3 and Tm5b only differ in their N terminal exons, but these N-terminal exons do not independently direct localisation within the cell, as chimeric mutants Tm3-Tm5NM1 and Tm5b-Tm5NM1 show an increased peripheral localisation similar to Tm5NM1. The lower abundance of Tm3 at the periphery of the cell is not a result of different protein dynamics, as Tm3 and Tm5b show similar recovery after photobleaching. The relative exclusion of Tm3 from the periphery of cells does, however, require interaction with the actin filament, as mutants with truncations at either the N-terminus or the C-terminus are unable to localise to actin stress fibres, and are present in the most peripheral regions of the cell. We conclude that it is the entire Tm molecule which is the unit of sorting, and that the alternatively spliced N terminal exons do not act as autonomous targeting signals. PMID- 20026408 TI - Curvature variation along the tropomyosin molecule. AB - Complementarity between the tropomyosin supercoil and the helical contour of actin-filaments is required for the binding interaction of actin and tropomyosin (Li et al., 2010). Clusters of small alanine residues in place of canonical leucines along coiled-coil tropomyosin may be responsible for pre-shaping tropomyosin and promoting conformational complementarity to F-actin. A longitudinal displacement between the two chains of the tropomyosin coiled-coil induced by the alanine clusters could produce localized bending or limited flexibility along tropomyosin needed to shape tropomyosin (Brown and Cohen, 2005). To evaluate the influence of alanine clusters on tropomyosin curvature, we calculated the longitudinal displacement between amino acid residues on adjacent chains of the tropomyosin coiled-coil and related this "Z-displacement" to the position of the alanine clusters. Measurements were made on high-resolution crystal structures of tropomyosin fragments and on trajectories from molecular dynamics simulations of full-length alphaalpha-tropomyosin. We found no strict one-for-one spatial correlation between alanine cluster position and the Z displacement. Neither did we find any direct correspondence between the clusters and the local curvature of tropomyosin. Rather than just causing specific local structural effects, the overall influence of alanine clusters is complex and delocalized, leading to a gradually changing bending pattern along the length of tropomyosin. PMID- 20026409 TI - Molecular cloning and expression of NOS in shrimp, Litopenaeus vannamei. AB - The importance of the nitric oxide synthase (NOS) gene family is demonstrated by many studies in recent years. However, the lack of sequence information and clones of shrimp NOS cDNA limits further study on its characterization and function in this species. In this report, the cDNA of NOS contained full-length ORF was cloned from the Pacific white shrimp, Litopenaeus vannamei. It was of 4680 bp, including a 5'-terminal untranslated region (UTR) of 278 bp, a 3' terminal UTR of 862 bp, which contained 5 ATTTA repeats, and an open reading frame (ORF) of 3540 bp encoding a polypeptide of 1179 amino acids. It contained a typical NO synthase domain at the N-terminal, next to a flavodoxin 1 domain, a flavin adenine dinucleotide (FAD) binding domain, respectively, and a conservative nicotinamide adenine dinucleotide (NAD) binding domain structure at the C-terminal. Quantitative real-time reverse transcription PCR analysis revealed L. vannamei NOS (LvNOS) to be expressed in most shrimp tissues, with highest expression in the hepatopancreas and weakest expression in skin. The expression of LvNOS after challenge with LPS and poly I:C was tested in hemocytes, hepatopancreas and nerve. The results indicated that the NOS transcript level could be induced in hemocytes by injection with LPS. The highest expression was in the hemocyte, with 8.8 times (at 3 h) as much as that in the control (p < 0.05). However, sharp down-regulation of NOS was found in hepatopancreas and nerve after LPS and poly I:C injection (p < 0.05). These results suggested that NOS might play an important role in shrimp's defense against pathogenic infection. PMID- 20026407 TI - An automated procedure for detecting protein folds from sub-nanometer resolution electron density. AB - The use of sub-nanometer resolution electron density as spatial constraints for de novo and ab initio structure prediction requires knowledge of protein boundaries to accurately segment the electron density for the prediction algorithms. Here we present a procedure where even poorly segmented density can be used to determine the fold of the protein. The method is automated, fast, capable of searching for multiple copies of a protein fold, and accessible to densities encompassing more than a thousand residues. The automation is particularly powerful as it allows the procedure to take full advantage of the expanding repository in the Protein Data Bank. We have tested the method on nine segmented sub-nanometer image reconstruction electron densities. The method successfully identifies the correct fold for the six densities for which an atomic structure is known, identifies a fold that agrees with prior structural data, a fold that agrees with predictions from the Fold & Function Assignment server, and a fold that correlates with secondary structure prediction. The identified folds in the last three examples can be used as templates for comparative modeling of the bacteriophage P22 tail-machine (a 3MDa complex composed of 39 protein subunits). PMID- 20026410 TI - Rediscovery of Protohynobius puxiongensis (Caudata: Hynobiidae) and its phylogenetic position based on complete mitochondrial genomes. AB - The mysterious Asian hynobiid salamander, Protohynobius puxiongensis, was described based on a single specimen collected in 1965 and never found again since then. Because the specimen had an internasal bone, Pr. puxiongensis was thought to retain a primitive character lost by a common ancestor of all other hynobiid salamanders, and it was thus considered to be not only a new genus, but also a new subfamily. This conclusion bothered herpetologists for decades because it was based on only one specimen and one character without other living specimens being rediscovered. After years of field effort, we rediscovered living individuals of Pr. puxiongensis at its type locality. All characters observed in rediscovered specimens are identical to the original description of the holotype except the internasal bone, implying that the internasal bone observed in the holotype may be just an individual variation. To examine the phylogenetic position of Pr. puxiongensis, we sequenced complete mitochondrial genomes for this species, together with two Pseudohynobius species. By combining 18 published hynobiid mitochondrial genomes and our new sequences, we reconstructed a comprehensive phylogenetic relationship of Hynobiidae at the genus level. Our results indicate that Pr. puxiongensis is deeply nested within the hynobiid phylogeny. It is the sister group of the Pseudohynobius species, and the validity of subfamily Protohynobiinae is not supported. PMID- 20026411 TI - A plastid tree can bring order to the chaotic generic taxonomy of Rytidosperma Steud. s.l. (Poaceae). AB - Rytidosperma s.l., wallaby grasses and allies, is in dire need of a single, unanimously accepted generic taxonomy. Motivated by the desire to establish a generic classification that complies with phylogeny, we investigated how much phylogenetic signal is contained within a plastid (cpDNA) tree, given that the nrDNA tree (ITS) was uninformative and that a phylogenetic hypothesis based on a single genome may not be reliable. We find that the plastid tree is significantly different from a morphological cladogram and show that this is the result of homoplasy in the morphological dataset. Treated individually, several morphological characters fit the plastid tree very well. Similarly, we find a good fit of the plastid tree with ecological and distribution characters and with biogeographical patterns in the Southern Hemisphere. We conclude that a significant level of the species phylogeny is resolved by the plastid tree and are confident it can form a sound basis for a reconsideration of generic limits. None of the currently recognised seven genera in the Rytidosperma clade is monophyletic. Therefore, we propose combining the segregate genera in Australasia within a broadly construed Rytidosperma, including all the species from Australia, New Guinea, New Zealand and South America. PMID- 20026412 TI - Neuronal representation of conditioned taste in the basolateral amygdala of rats. AB - Animals develop robust learning and long lasting taste aversion memory once they experience a new taste that is followed by visceral discomfort. A large body of literature has supported the hypothesis that basolateral amygdala (BLA) plays a critical role in the acquisition and extinction of such conditioned taste aversions (CTA). Despite the evidence that BLA is crucially engaged during CTA training, it is unclear how BLA neural activity represents the conditioned tastes. Here, we incorporated a modified behavioral paradigm suitable for single unit study, one which utilizes a sequence of pulsed saccharin and water infusion via intraoral cannulae. After conditioning, we investigated BLA unit activity while animals experience the conditioned taste (saccharin). Behavioral tests of taste reactivity confirmed that the utilized training procedure produced reliable acquisition and expression of the aversion throughout test sessions. When neural activity was compared between saccharin and water trials, half of the recorded BLA units (77/149) showed differential activity according to the types of solution. 76% of those cells (29/38) in the conditioned group showed suppressed activity, while only 44% of taste reactive cells (17/39) in controls showed suppressed activity during saccharin trials (relative to water trials). In addition, the overall excitability of BLA units was increased as shown by altered characteristics of burst activity after conditioning. The changes in BLA activity as a consequence of CTA were maintained throughout test sessions, consistent with the behavioral study. The current study suggests that the neuronal activity evoked by a sweet taste is altered as a consequence of CTA learning, and that the overall change might be related to the learning induced negative affect. PMID- 20026413 TI - Late-onset hemorrhagic cystitis in children after hematopoietic stem cell transplantation for thalassemia and sickle cell anemia: a prospective evaluation of polyoma (BK) virus infection and treatment with cidofovir. AB - Little is known about late-onset hemorrhagic cystitis (HC) in children, its relationship to BK virus, and treatment with cidofovir (CDV) following hematopoietic stem cell transplantation (HSCT). We prospectively investigated BK virus reactivation in children who underwent HSCT from a matched related donor for thalassemia or sickle cell anemia following busulfan-cyclophosphamide-based conditioning regimens and analyzed risk factors for development of HC and its treatment with CDV. Grade 2-4 HC occurred in 30 patients with a cumulative incidence of 26% (95% confidence interval [CI] = 18%-34%). The cumulative incidences of BK viruria and viremia were 81% (95% CI = 69%-89%) and 28% (95% CI = 18%-40%), respectively. Multivariate analysis revealed that use of antithymocyte globulin (ATG) (hazard ratio [HR] = 10.5; P = .001), peak BK viruria >100,000 copies/mL (HR = 6.2; P = .004), and grade II-IV acute graft versus-host disease (HR = 5.3; P = .007) were predictive factors for HC. Nineteen patients with HC were given CDV at 1.5 mg/kg/day 3 times a week, or 5 mg/kg/week. The median duration of therapy was 27 days (range, 21-180 days), and a median of 9 doses were given (range, 6-22). All patients had a complete clinical response (CCR), and 69% had a microbiological response at 4 weeks. Eleven patients with BK virus-related HC receiving supportive care also had CCR. The median duration of HC in these patients was similar to that in patients treated with CDV. None of the patients with HC cleared BK viruria when CCR was achieved. We conclude that late-onset HC is more prevalent in children with sustained high BK viruria who are treated with ATG or who develop graft-versus-host disease. Randomized clinical trials are urgently needed to better define the role of CDV in treating BK virus-related HC. PMID- 20026414 TI - ERAD substrates: which way out? AB - Global folding of polypeptides entering the endoplasmic reticulum (ER) starts as soon as they emerge from the narrow Sec61 translocon. Attainment of the native structure can take from several minutes to hours, depending on the gene product. Until then, non-native folding intermediates must be protected from molecular chaperones that recognize misfolded determinants and could prematurely interrupt folding programs by re-directing them to disposal pathways. On the other hand, futile folding attempts must actively be stopped to prevent intraluminal accumulation of defective cargo. This review describes recent advances in understanding how terminally misfolded polypeptides are extracted from the folding environment and directed to specific dislocons within the ER membrane for transfer to the cytoplasm for proteasome-mediated degradation. PMID- 20026415 TI - An expanded genetic linkage map of an intervarietal Agaricus bisporus var. bisporusxA. bisporus var. burnettii hybrid based on AFLP, SSR and CAPS markers sheds light on the recombination behaviour of the species. AB - A genetic linkage map for the edible basidiomycete Agaricus bisporus was constructed from 118 haploid homokaryons derived from an intervarietal A. bisporus var. bisporus x A. bisporus var. burnettii hybrid. Two hundred and thirty-one AFLP, 21 SSR, 68 CAPS markers together with the MAT, BSN, PPC1 loci and one allozyme locus (ADH) were evenly spread over 13 linkage groups corresponding to the chromosomes of A. bisporus. The map covers 1156cM, with an average marker spacing of 3.9cM and encompasses nearly the whole genome. The average number of crossovers per chromosome per individual is 0.86. Normal recombination over the entire genome occurs in the heterothallic variety, burnettii, contrary to the homothallic variety, bisporus, which showed adaptive genome-wide suppressed recombination. This first comprehensive genetic linkage map for A. bisporus provides foundations for quantitative trait analyses and breeding programme monitoring, as well as genome organisation studies. PMID- 20026416 TI - Plasticity in body temperature and metabolic capacity sustains winter activity in a small endotherm (Rattus fuscipes). AB - Small mammals that remain active throughout the year at a constant body temperature have a much greater energy and food requirement in winter. Lower body temperatures in winter may offset the increased energetic cost of remaining active in the cold, if cellular metabolism is not constrained by a negative thermodynamic effect. We aimed to determine whether variable body temperatures can be advantageous for small endotherms by testing the hypothesis that body temperature fluctuates seasonally in a wild rat (Rattus fuscipes); conferring an energy saving and reducing food requirements during resource restricted winter. Additionally we tested whether changes in body temperature affected tissue specific metabolic capacity. Winter acclimatized rats had significantly lower body temperatures and thicker fur than summer acclimatized rats. Mitochondrial oxygen consumption and the activity of enzymes that control oxidative (citrate synthase, cytochrome c-oxidase) and anaerobic (lactate dehydrogenase) metabolism were elevated in winter and were not negatively affected by the lower body temperature. Energy transfer modeling showed that lower body temperatures in winter combined with increased fur thickness to confer a 25 kJ day(-1) energy saving, with up to 50% owing to reduced body temperature alone. We show that phenotypic plasticity at multiple levels of organization is an important component of the response of a small endotherm to winter. Mitochondrial function compensates for lower winter body temperatures, buffering metabolic heat production capacity. PMID- 20026418 TI - SNPs in the myostatin gene of the mollusk Chlamys farreri: association with growth traits. AB - Myostatin (MSTN) is a member of the transforming growth factor-beta superfamily which negatively regulates growth of muscle tissue. In this study, 103 cultivated Chlamys farreri individuals were screened for polymorphisms in the MSTN gene using PCR-single strand conformation polymorphism (PCR-SSCP) and DNA sequencing methods. Two mutations were found: A/G at position 327 in exon 2, which caused an amino acid change from Thr to Ala (Thr305Ala), and C/T at position 289 in exon 3, which caused an amino acid change from Cys to Arg (Cys422Arg). One way ANOVA of the SNPs and growth traits showed that genotype GG of primer M5 had significantly higher body mass, soft-tissue mass, adductor muscle mass, shell length, shell height, absolute growth rate of shell height and body mass than those of genotype AG and AA (P<0.05). Genotype frequencies of genotype AA, AG and GG were 68.94%, 27.18% and 3.88%, respectively. The results present evidence that the C. farreri MSTN gene may be selected as a candidate gene for these growth traits. PMID- 20026417 TI - Melatonin modulates glucose homeostasis during winter dormancy in a vespertilionid bat, Scotophilus heathi. AB - The role for melatonin in glucose homeostasis and insulin resistance is not very clear and has recently been an active area of investigation. The present study investigated the role of melatonin in seasonal accumulation of adipose tissue in Scotophilus heathi, with particular reference to its role in glucose homeostasis and development of insulin resistance. The circulating melatonin levels correlated positively (p<0.05) with the changes in body mass due to fat accumulation and circulating insulin level, but correlated negatively with the blood glucose level in S. heathi. The bats showed high circulating blood glucose levels and impaired glucose tolerance during the period of fat deposition suggesting insulin resistance condition which improves after winter when most of the fat has been utilized as a metabolic fuel. The high circulating melatonin levels during the period of maximum body fat at the beginning of winter prepare the bats for winter dormancy by modulating the glucose homeostasis through affecting blood glucose levels, muscle and liver glycogen stores, insulin receptor and glucose transporter 4 (GLUT 4) expression. This is also confirmed by in vivo study in which melatonin injection improves the glucose tolerance, increases muscle insulin receptor and GLUT 4 expression, and enhances glucose clearance from the blood. The results of present study further showed that the effect of melatonin injection on the blood glucose levels is determined by the metabolic state of the bats and may protect from decrease in blood glucose level during extreme starvation, however, melatonin when injected during fed state increases glucose clearance from the blood. In summary, the present study suggested that melatonin interferes with the glucose homeostasis through modulating intracellular glucose transport and may protect bats from hypoglycemia during winter dormancy. PMID- 20026419 TI - Transporters, channels, or simple diffusion? Dogmas, atypical roles and complexity in transport systems. AB - The recent breakthrough discoveries of transport systems assigned with atypical functions provide evidence for complexity in membrane transport biochemistry. Some channels are far from being simple pores creating hydrophilic passages for solutes and can, unexpectedly, act as enzymes, or mediate high-affinity uptake, and some transporters are surprisingly able to function as sensors, channels or even enzymes. Furthermore, numerous transport studies have demonstrated complex multiphasic uptake kinetics for organic and mineral nutrients. The biphasic kinetics of glucose uptake in Saccharomyces cerevisiae, a result of several genetically distinct uptake systems operating simultaneously, is a classical example that is a subject of continuous debate. In contrast, some transporters display biphasic kinetics, being bona fidae dual-affinity transporters, their kinetic properties often modulated by post-translational regulation. Also, aquaporins have recently been reported to exhibit diverse transport properties and can behave as highly adapted, multifunctional channels, transporting solutes such as CO(2), hydrogen peroxide, urea, ammonia, glycerol, polyols, carbamides, purines and pyrimidines, metalloids, glycine, and lactic acid, rather than being simple water pores. The present review provides an overview on some atypical functions displayed by transporter proteins and discusses how this novel knowledge on cellular uptake systems may be related to complex multiphasic uptake kinetics often seen in a wide variety of living organisms and the intriguing diffusive uptake of sugars and other solutes. PMID- 20026420 TI - The resolution of inflammation: anti-inflammatory roles for NF-kappaB. AB - Inflammation is a salutary response to insult or injury that normally resolves with no detriment to the host. While the mechanisms and mediators that regulate the onset of inflammation have been well characterized we still know relatively little about the endogenous mechanisms that terminate the inflammatory response (Lawrence and Gilroy, 2007). Nuclear factor (NF)-kappaB is a generic term for a family of ubiquitous transcription factors with diverse physiological functions (Bonizzi and Karin, 2004; Caamano and Hunter, 2002). NF-kappaB transcription factors are formed by dimerisation of Rel proteins; RelA (p65), c-Rel, RelB, p50, p52. Various hetero or homodimers of Rel proteins can be formed in a tissue and stimulus specific manner, genetic evidence suggests these transcription factors have a critical role in cell survival and pro-inflammatory signalling pathways, which have been extensively reviewed elsewhere (Bonizzi and Karin, 2004; Caamano and Hunter, 2002). The critical role for NF-kappaB in pro-inflammatory gene expression has led to an enormous effort to develop inhibitors of this pathway for the treatment of chronic inflammation (Karin et al., 2004). However, recent research using modern molecular genetic approaches has revealed new anti inflammatory roles for NF-kappaB that may have important implications for targeting this pathway in the treatment of inflammatory diseases. In this review we will discuss the emerging role of NF-kappaB in the resolution of inflammation and some of the potential mechanisms attributed to this function. PMID- 20026421 TI - The complexity of the complicity of mast cells in cancer. AB - Mast cells are evolutionarly ancient cells of the immune cells which can secrete a variety of effector molecules. Animal and pathologic studies suggest that mast cells may promote tumor growth in some cancer types but may act in an opposite manner in others. In several mouse models a critical role of mast cells for tumor promotion was demonstrated. In humans mast cells are dependent upon the tyrosine kinase receptor c-Kit. This receptor is inhibited by many of the new anti-cancer tyrosine kinase inhibitors including Pazopanib, Imatinib and Masitinib. These drugs probably ablate some tumor mast cells, in addition to their other known antitumor effects. Understanding the complex roles of mast cells in cancer should aid in understanding mechanisms of current tyrosine kinase inhibitors, and the development of innovative anti-cancer therapies. PMID- 20026422 TI - MicroRNAs: potential biomarkers for cancer diagnosis, prognosis and targets for therapy. AB - MicroRNAs have a revolutionary impact on cancer research over recent years. They emerge as important players in tumorigenesis, leading to a paradigm shift in oncology. The widespread and comprehensive use of microRNA microarrays has enabled the identification of a number of microRNAs as potential biomarkers for cancer. It is encouraging to report that microRNAs have remarkable stability in both formalin-fixed tissue and blood. Many microRNAs have been identified to act as oncogenes, tumor suppressors, or even modulators of cancer stem cells and metastasis. Some studies not only reported the identified microRNA biomarkers, but also deciphered their target genes and the underlying mechanisms. The rapid discovery of many microRNA targets and their relevant pathways has contributed to the development of microRNA-based therapeutics, but the developing progress of antisense or siRNA drugs has been hampered by stability, specificity and delivery problems. This review summarizes the most significant and latest findings of original researches on microRNAs involvement in cancer, focusing on the potential of cancer-related microRNAs as biomarkers for diagnosis, prognosis and targets for therapy. PMID- 20026423 TI - Eicosanoids and the endogenous control of acute inflammatory resolution. AB - Inflammation is a formidable ally in the constant battle against infection, cancer and tissue injury. It is a primordial response that protects against injury and restores damaged tissue to its normal physiological functioning. In fact, our wellbeing and survival depends upon its efficiency and carefully balanced control. In general, the innate inflammatory response initiates within minutes and, if all is well, resolves within hours. In contrast, chronic inflammation persists for weeks, months or even years. Here, we are going to discuss the key endogenous checkpoints necessary for mounting an effective yet limited inflammatory response and the crucial biochemical pathways necessary to prevent its persistence. In this setting, the biochemical synthesis of key pro resolution eicosanoids as well as their mode of action in self-limiting inflammation will be discussed. PMID- 20026424 TI - In vitro investigations of glycidamide-induced DNA lesions in mouse male germ cells and in mouse and human lymphocytes. AB - The industrial compound and food contaminant acrylamide (AA) is a probable human carcinogen, also known to induce male-mediated reproductive effects in animals. Most data suggest that its metabolite glycidamide (GA) is involved in the observed toxicity. We have investigated in vitro effects of AA/GA in mouse male germ cells (prior to spermatid elongation) and human and mouse peripheral blood lymphocytes, to assess inter-species and cell-type differences in susceptibility, and to explore the nature of the DNA lesion(s) as well as their potential repair. The comet assay was used in combination with the DNA-repair enzymes Fpg and hOGG1 to measure specific DNA lesions. In contrast to AA, GA induced significant levels of DNA lesions (strand breaks and alkali-labile sites) at millimolar concentrations in mouse testicular cells and human peripheral blood lymphocytes (hPBL). Using Fpg, the GA-induced DNA damage was measured at 20-50-fold higher sensitivity, in all cell types investigated. GA-induced DNA damage could not be recognised by hOGG1, suggesting that, based on the known affinities of these repair enzymes, alkylation of guanine is involved, rather than oxidation. Human lymphocytes appeared to be more susceptible to GA-induced lesions than both types of mouse cells. Mouse testicular cells and lymphocytes seemed to respond similarly to GA-induced Fpg-sensitive DNA lesions. The persistence of lesions was explored with cells from mice either proficient or deficient in Ogg1 (mouse 8 oxoguanine DNA glycosylase). Low in vitro repair of GA-induced Fpg-sensitive lesions was observed in primary male germ cells and lymphocytes from both Ogg1(+/+) and Ogg1(-/-) mice. We conclude that there may be differences between mice and humans in AA/GA-induced genotoxicity, and DNA from mouse male germ cells does not appear to be more sensitive to GA than DNA from peripheral blood lymphocytes in vitro. The usefulness of the comet assay in combination with DNA repair enzymes is demonstrated. PMID- 20026425 TI - Zschokkella hildae Auerbach, 1910: phylogenetic position, morphology, and location in cultured Atlantic cod. AB - The myxozoan Zschokkella hildae Auerbach, 1910, was detected with a prevalence of 100% in cultured Atlantic cod, Gadus morhua L. aged 1+ from a culture facility on the west coast of Scotland. Sporogonic stages of Z. hildae, plasmodia producing 2 5 mature spores, were located predominantly in the collecting ducts and ureters of the kidney, and spores were present in the urine collected from the bladder. Less frequently, plasmodia were detected in the interstitial tissue of the kidney. The parasite prevalence in cultured fish was considerably higher than reported in wild fish but no obvious signs of pathology were detected. SSU rDNA sequencing and phylogenetic analysis showed that Z. hildae is closely related to a Sinuolinea sp. from the urinary system of turbot, Psetta maxima (L.), and that these two species, together with other myxozoans from the urinary system of marine fish cluster together in a sub-clade of the recognised marine clade of myxozoans. This sub-clade is characterised by a specific linear expansion segment, helix E23_15 in the secondary structure of variable region V4 of the SSU rDNA. Z. hildae and Sinuolinea sp. show extraordinary large linear expansion segment in both V4 and V7 and an important number of complementary base changes in the conservative regions of the SSU rDNA, indicating considerable evolutionary changes in the SSU rDNA of these species when compared with other myxozoans from the marine environment. PMID- 20026426 TI - Differential sensitivities of pulmonary and coronary arteries to hemoglobin-based oxygen carriers and nitrovasodilators: study in a bovine ex vivo model of vascular strips. AB - Vasoconstriction is a major adverse effect of first and second generation hemoglobin-based oxygen carriers (HBOCs) that hinders their development as blood substitute. However, intravenous infusion of HBOC-201 (second generation) to patients induces significant pulmonary hypertension without significant coronary vasoconstriction. We compared contractile responses of isolated bovine pulmonary and coronary arterial strips to HBOC-201 and HBOC-205LL.LT.MW600 (third generation), polymerized bovine hemoglobins of different molecular weight, and their attenuation by nitroglycerin, sodium nitroprusside (SNP), and sodium nitrite. Pulmonary arteries developed negligible basal tone, but exhibited HBOC dependent amplification of phenylephrine-induced contractions. In contrast, coronary arteries developed significant basal tone, and exhibited HBOC-dependent constant force increment to serotonin-induced contractions. Therefore, relative to basal tone, HBOC-induced contractions were greater in pulmonary than coronary arteries. Furthermore, HBOC-205LL.LT.MW600 appeared to be less vasoactive than HBOC-201. Unexpectedly, pulmonary and coronary arteries exhibited differential sensitivities to nitrovasodilators in parallel with their differential sensitivities to HBOC. However, SNP and sodium nitrite induced significant methemoglobin formation from HBOC, whereas nitroglycerin did not. These results suggest that phenotypic differences between pulmonary and coronary vascular smooth muscle cells could explain the differential hypertensive effects of HBOC on pulmonary and coronary circulation in patients. Among the three nitrovasodilators investigated, nitroglycerin appears to be the most promising candidate for attenuating HBOC-induced pulmonary hypertension in older HBOCs. PMID- 20026427 TI - Update on colonoscopic imaging and projections for the future. PMID- 20026428 TI - Limited genetic polymorphism of the Plasmodium vivax low molecular weight rhoptry protein complex in the Colombian population. AB - Proteins involved in parasite adhesion and invasion are considered the best candidates for the development of asexual blood-stage antimalarial vaccines. Such vaccine candidates should be accessible by the immune system and have limited diversity. Considering the promising results obtained in previous trials by immunizing monkeys with the rhoptry-associated proteins 1 and 2 (RAP-1 and RAP 2), here we assessed the genetic variability of the Plasmodium vivax rap-1 and rap-2 genes isolated from Colombian parasite populations. Limited sequence diversity was found in these genes, possibly as a result of a functional/structural restriction. The presence of several haplotypes at relatively low frequencies and the excess of singleton mutations suggests that a demographic process might be affecting the loci. Our results support the inclusion of PvRAP-1 and PvRAP-2 in the design of an antimalarial subunit-based vaccine against P. vivax, which would avoid inducing allele-specific immunity. PMID- 20026429 TI - Wing shape of dengue vectors from around the world. AB - Wing shape is increasingly utilized in species identification and characterization. For dengue vectors Aedes aegypti and Aedes albopictus, it could be used as a complement for ensuring accurate diagnostic of damaged specimens. However, the impact of world migration on wing shape is unknown. Has the spread of these invasive species increased shape variation to the extent of producing interspecific overlapping? To answer this question, the geometric patterns of wing venation in Ae. aegypti and Ae. albopictus were compared between natural populations from the Pacific Islands, North and South America and South East Asia. The geometry of 178 female and 174 male wings were described at 13 anatomical landmarks, and processed according to Procrustes superposition, partial warps and subsequent multivariate analyzes. The variation of shape did not produce significant interspecific overlapping. Regardless of geographic origin, Ae. aegypti was recognized as Ae. aegypti and Ae. albopictus as Ae. albopictus. Some significant geographic differentiation was observed in Colombia for Ae. aegypti and in Thailand for Ae. albopictus. Globally, the morphology of these mosquitoes, for both size and shape, appeared well preserved. Strong canalizing mechanisms could account for the observed patterns of relatively uniform morphology, which could also be attributed to sporadic, recurrent mixing of populations, thwarting phenotypic drift. PMID- 20026430 TI - Polymyositis, dermatomyositis and malignancy: a further intriguing link. AB - The association between malignancy and autoimmune myositis has been largely described and confirmed by numerous epidemiological studies. The temporal relationship between the two pathologic conditions can vary: malignancy may occur before, at the same time or following the diagnosis of myositis. Beside these observations, the molecular mechanisms underlying this association are still unknown, even though it has been demonstrated a possible antigenic similarity between regenerating myoblasts and some cancer cell populations. To better identify peculiar histopathologic features common to cancer and myositis, we screened muscle biopsies from patients affected with polymyositis, dermatomyositis, myositis in association to cancer, and from patients affected with newly diagnosed cancer, but without myositis. Similarly to the histopatologic features that were observed in the muscle from myositis patients, especially in those with cancer associated myositis, in patients affected with malignancy at the clinical onset of disease we observed early sign of myopathy, characterized by internally nucleated and regenerating myofibers, most of them expressing the neural cell adhesion molecule. The hypothesis that in a particular subset of individuals genetically predisposed to autoimmunity, an initial subclinical tumor-induced myopathy may result in an autoimmune myositis, represents a further intriguing link behind the association of these two conditions. PMID- 20026431 TI - Role of locus coeruleus noradrenergic neurons in cardiorespiratory and thermal control during hypoxia. AB - We investigated the specific role of LC noradrenergic (NA) neurons in the cardiorespiratory and thermal responses to hypoxia. Aiming this, LC NA neurons were selectively destroyed by microinjecting 6-OHDA bilaterally in the LC of male rats. Pulmonary ventilation (VE, plethysmograph), core body temperature (Tc, dataloggers), mean arterial pressure (MAP) and heart rate (fH) were measured during normoxia followed by 7% O(2) in air. Tyrosine hydroxylase (TH) immunoreactivity was performed to verify the chemical lesions effectiveness. Hypoxia caused increase in VE, decrease in Tc and did not affect fH in both sham operated and 6-OHDA-lesioned groups. In addition, hypoxia decreased MAP in the sham-operated group and did not affect MAP in the 6-OHDA-lesioned group. LC lesion did not affect VE, MAP, fH and Tc. Thus, these results indicate that LC noradrenergic neurons do not play a role in cardiorespiratory control and thermoregulation under basal and hypoxic conditions. PMID- 20026432 TI - Divorcing folding from function: how acylation affects the membrane-perturbing properties of an antimicrobial peptide. AB - Many small cationic peptides, which are unstructured in aqueous solution, have antimicrobial properties. These properties are assumed to be linked to their ability to permeabilize bacterial membranes, accompanied by the transition to an alpha-helical folding state. Here we show that there is no direct link between folding of the antimicrobial peptide Novicidin (Nc) and its membrane permeabilization. N-terminal acylation with C8-C16 alkyl chains and the inclusion of anionic lipids both increase Nc's ability to form alpha-helical structure in the presence of vesicles. Nevertheless, both acylation and anionic lipids reduce the extent of permeabilization of these vesicles and lead to slower permeabilization kinetics. Furthermore, acylation significantly decreases antimicrobial activity. Although acyl chains of increasing length also increase the tendency of the peptides to aggregate in solution, this cannot rationalize our results since permeabilization and antimicrobial activities are observed well below concentrations where aggregation occurs. This suggests that significant induction of alpha-helical structure is not a prerequisite for membrane perturbation in this class of antimicrobial peptides. Our data suggests that for Nc, induction of alpha-helical structure may inhibit rather than facilitate membrane disruption, and that a more peripheral interaction may be the most efficient permeabilization mechanism. Furthermore, acylation leads to a deeper embedding in the membrane, which could lead to an anti-permeabilizing "plugging" effect. PMID- 20026433 TI - Structural and mechanistic underpinnings of the differential drug sensitivity of EGFR mutations in non-small cell lung cancer. AB - EGFR and other ErbB-family tyrosine kinases are overexpressed in many human tumors, and their aberrant expression and mutational activation is associated with the development, progression and aggressiveness of a number of malignancies. Thus the EGFR kinase has long been recognized as a potential drug target in oncology, and small-molecule inhibitors have been under development for more than two decades. As a result of their effectiveness in treating non-small cell lung cancers (NSCLCs) driven by somatic mutations in the EGFR kinase, gefitinib and erlotinib were the first EGFR tyrosine kinase inhibitors (TKIs) approved for clinical use. Ironically, these drugs found their target against mutant forms of the EGFR kinase, which have altered enzyme active sites, and not against the wild type (WT) kinase against which their potency and selectivity was carefully honed. Here we review recent structural and enzymological studies that explore the exquisite sensitivity of a subset of these lung cancer mutants to gefitinib and erlotinib. We discuss available structural evidence for the mechanisms of activation of the EGFR kinase by these mutants, and compare it to physiologic activation of the kinase by ligand-induced dimerization. Finally, we consider the mechanisms by which the secondary T790M "gatekeeper" mutation confers resistance to gefitinib and erlotinib. PMID- 20026435 TI - Competitive protein adsorption on biomaterial surface studied with reflectometric interference spectroscopy. AB - Reflectometry interference spectroscopy (RIfS) is known as a highly sensitive and robust technique for direct, label-free detection of the interaction of biomacromolecules in real time and in situ. The aim of the present study was to investigate the competitive protein adsorption on the surface of fluorocarbon end capped poly(carbonate) urethane (PCUF) and polystyrene (PS) based on the RIfS method. The surface energy and microstructures of PCUF and PS were characterized by contact angle measurement and atomic force microscopy. Interfacial energies between these surfaces and the proteins were then calculated. The protein adsorption experiments were carried out with both single solution and ternary solutions composed of albumin, fibrinogen and immunoglobulin-G (IgG). The results of surface characterization showed that PCUF was more hydrophilic than PS with a smaller surface energy, and micro-phases separation of PCUF was observed. RIfS analysis results revealed that more albumins, less fibrinogen and IgG were detected on the PCUF surface compared with PS after simplex and competitive protein adsorption, which indicated that PCUF had a preferential adsorption for albumin. The special morphology, smaller surface energy and calculated interfacial energies between PCUF and proteins may be responsible for the better blood compatibility of PCUF compared to PS. The results suggest that RIfS could serve as a novel, effective method for studying the competitive protein adsorption on biomaterial surfaces. PMID- 20026434 TI - Modulation of advanced glycation endproduct synthesis by kynurenines in human lens proteins. AB - Human lens proteins (HLP) become chemically modified by kynurenines and advanced glycation end products (AGEs) during aging and cataractogenesis. We investigated the effects of kynurenines on AGE synthesis in HLP. We found that incubation with 5 mM ribose or 5 mM ascorbate produced significant quantities of pentosidine, and this was further enhanced in the presence of two different kynurenines (200-500 microM): N-formylkynurenine (Nfk) and kynurenine (Kyn). Another related compound, 3-hydroxykynurenine (3OH-Kyn), had disparate effects; low concentrations (10-200 microM) promoted pentosidine synthesis, but high concentrations (200-500 microM) inhibited it. 3OH-Kyn showed similar effects on pentosidine synthesis from Amadori-enriched HLP or ribated lysine. Chelex-100 treatment of phosphate buffer reduced pentosidine synthesis from Amadori-enriched HLP by approximately 90%, but it did not inhibit the stimulating effect of 3OH-Kyn and EDTA. 3OH-Kyn (100-500 microM) spontaneously produced copious amounts of H(2)O(2) (10-25 microM), but externally added H(2)O(2) had only a mild stimulating effect on pentosidine but had no effect on N(epsilon)-carboxymethyl lysine (CML) synthesis in HLP from ribose and ascorbate. Further, human lens epithelial cells incubated with ribose and 3OH-Kyn showed higher intracellular pentosidine than cells incubated with ribose alone. CML synthesis from glycating agents was inhibited 30 to 50% by 3OH Kyn at concentrations of 100-500 microM. Argpyrimidine synthesis from 5mM methylglyoxal was slightly inhibited by all kynurenines at concentrations of 100 500 microM. These results suggest that AGE synthesis in HLP is modulated by kynurenines, and such effects indicate a mode of interplay between kynurenines and carbohydrates important for AGE formation during lens aging and cataract formation. PMID- 20026436 TI - Porous polycaprolactone scaffold for cardiac tissue engineering fabricated by selective laser sintering. AB - An advanced manufacturing technique, selective laser sintering (SLS), was utilized to fabricate a porous polycaprolactone (PCL) scaffold designed with an automated algorithm in a parametric library system named the "computer-aided system for tissue scaffolds" (CASTS). Tensile stiffness of the sintered PCL strut was in the range of 0.43+/-0.15MPa when a laser power of 3W and scanning speed of 150 in s(-1) was used. A series of compressive mechanical characterizations was performed on the parametric scaffold design and an empirical formula was presented to predict the compressive stiffness of the scaffold as a function of total porosity. In this work, the porosity of the scaffold was selected to be 85%, with micropores (40-100mum) throughout the scaffold. The compressive stiffness of the scaffold was 345kPa. The feasibility of using the scaffold for cardiac tissue engineering was investigated by culturing C2C12 myoblast cells in vitro for 21days. Fluorescence images showed cells were located throughout the scaffold. High density of cells at 1.2x10(6)cellsml(-1) was recorded after 4days of culture. Fusion and differentiation of C2C12 were observed as early as 6days in vitro and was confirmed with myosin heavy chain immunostaining after 11days of cell culture. A steady population of cells was then maintained throughout 21days of culturing. This work demonstrated the feasibility of tailoring the mechanical property of the scaffold for soft tissue engineering using CASTS and SLS. The macroarchitecture of the scaffold can be modified efficiently to fabricate scaffolds with different macropore sizes or changing the elemental cell design in CASTS. Further process and design optimization could be carried out in the future to fabricate scaffolds that match the tensile strength of native myocardium, which is of the order of tens of kPa. PMID- 20026437 TI - Assessment of biocompatibility and initial evaluation of genipin cross-linked elastin-like polypeptides in the treatment of an osteochondral knee defect in rabbits. AB - Polypeptides based on the alternating hydrophobic and cross-linking domain structure of human elastin are capable of undergoing self-assembly to produce polymeric matrices with unique biological and mechanical properties. Here, we test the initial feasibility of using a genipin cross-linked elastin-based material as an acellular plug in the treatment of an osteochondral defect in the rabbit knee. Full-thickness defects in the weight-bearing surface of the medial femoral condyle in 18 New Zealand White rabbits were surgically produced and press fitted with cylindrical pads composed of genipin cross-linked elastin-like polypeptides, with identical wounds in the opposite knee left untreated as controls. The biocompatibility of the material, overall wound healing and regeneration of subchondral tissue was assessed at 2, 4 and 6weeks by histological evaluation, synovial fluid analysis and microcomputerized tomography scanning. Histological analysis revealed the regeneration of subchondral bone at the periphery of the material, with evidence of hyaline-like overgrowth across the apical surface in 11/16 cases. Pads developed tight contacts with host tissue and appeared completely biocompatible, with no evidence of localized immune response or increased inflammation compared to controls. The material was stable to 6weeks, with an aggregate elastic modulus calculated at approximately 470kPa when tested under confined compression. Further studies are required to assess material degradation over time and long-term replacement with repair tissue. PMID- 20026438 TI - Uptake and distribution of labeled antibodies into pH-sensitive microgels. AB - We investigated the uptake and release of labeled antibodies from pH-sensitive hydrogel microparticles (i.e. microgels) by means of fluorescence analysis of labeled biological samples. The poly(methacrylic acid) (PMAA) hydrogel is a carbon-based network having carboxylic groups on the surface that dissociate according to their acid-base equilibrium. The ability of the PMAA microgel to encapsulate and release anti-CD4 and anti-CD8 monoclonal antibodies (MAbs), differing for the isotype and labeled with highly photostable fluorophore, was studied in solution by photoluminescence spectroscopy. The experimental results indicated that the uptake and release of the tested antibodies were controlled by pH. Furthermore, confocal microscopy analysis in the solid state revealed that the distribution of the labeled antibodies either on the surface or in the core of the microgel matrix was related to the specific properties of these MAbs. PMID- 20026439 TI - Microstructures and mechanical properties of silks of silkworm and honeybee. AB - Natural silks as important biomaterials have wide applications in scaffolds for tissues, biocompatible coatings and drug delivery, etc. In this paper, we report on the microstructures and mechanical properties of honeybee and silkworm silks. The microstructures and mechanical properties of these natural silks are measured by environment scanning electron microscopy (ESEM), scanning probe microscopy (SPM), tensile tests, and nanoindentation. It is found that the honeybee silk appears as a single fiber with a circular cross-section and has a much finer and smoother texture than the silkworm silk. The honeybee silk exhibits a distinct linear and brittle elastic feature with an elastic modulus of 6.5GPa and a breaking strain of 3.8%, whereas the silkworm silk possesses a nonlinear and hierarchical failure character with an initial elastic modulus of 8.9GPa and a breaking strain of 15.7%. Moreover, the nanoindentation measurements show that the honeybee silk exhibits much less anisotropy than the silkworm silk. The ratio of the longitudinal modulus to the transverse modulus of the honeybee silk is 2.0, whereas that of the silkworm silk is 18.9. The different structural and mechanical properties of the honeybee and silkworm silks are likely a result of their specific biological functions. PMID- 20026440 TI - Comparison of effects of alfacalcidol and alendronate on mechanical properties and bone collagen cross-links of callus in the fracture repair rat model. AB - Both bone density and quality are important determinants of bone strength. Bone quality is prescribed by matrix characteristic including collagen cross-linking and bone structural characteristics and is important in reinforcement of bone strength. We investigated the effects of alfacalcidol (ALF), a prodrug of calcitriol, and alendronate (ALN), a bisphosphanate, on the mechanical properties and content of enzymatic cross-links in femoral bone using a fracture repair rat model. Forty 3-month-old female Wistar-Imamichi rats were randomized into 4 groups: SHAM (sham-operated+vehicle), OVX (ovariectomy+vehicle), ALF (ovariectomy+ALF, 0.1 microg/kg/d, p.o.) and ALN (ovariectomy+ALN, 10 microg/kg/d, s.c.). Treatment began immediately after SHAM or OVX surgery. Three weeks later, all animals underwent transverse osteotomies at the midshaft of the left femur. Treatment was continued and rats were sacrificed at 12 weeks post fracture for evaluation by X-ray radiography, micro-CT, pQCT, biomechanical testing and bone histomorphometry. In the ALN group, no new cortical shell appeared and the callus diameter was significantly larger than in the OVX group (p<0.05). Stiffness of fractured callus in the ALF group, but not in the ALN group, was significantly higher than in the OVX group. Young's modulus in the ALN group was significantly decreased compared to the OVX group. Moreover, micro-CT analysis showed that ALN treatment increased the lowly mineralized bone in the callus by, resulting in the highest content of woven bone area and lowest content of lamellar bone. The total amount of enzymatic cross-links in both the ALF and ALN groups was significantly higher than in the OVX control group. Of particular interest, the Pyr-to-Dpyr ratio was significantly decreased by ALF administration, suggesting that ALF but not ALN normalized the enzymatic cross link patterns in fractured bone to the control level. In conclusion, ALN and ALF treatment increased bone strength via the distinctive effect on bone mass and quality. ALN formed larger calluses and increased enzymatic cross-links despite delayed woven bone remodeling into lamellar bone, whereas ALF treatment induced lamellar bone formation coincided with increasing in the enzymatic cross-linking and normalizing the cross-link pattern in callus to native bone pattern. PMID- 20026441 TI - Role of viral factors in modifying weight-related effects on disease progression of chronic hepatitis C patients. PMID- 20026442 TI - Recurrent nausea, vomiting, and abdominal pain. PMID- 20026445 TI - The American College of Gastroenterology irritable bowel syndrome monograph: translating systematic review data to clinical practice. PMID- 20026447 TI - Esophageal blebs and blisters. PMID- 20026448 TI - Optical molecular imaging approaches in colorectal cancer. PMID- 20026449 TI - Oral immunization with a novel lipid-based adjuvant protects against genital Chlamydia infection. AB - Oral immunization is attractive as a delivery route because it is needle-free and useful for rapid mass vaccination programs to target pandemics or bioterrorism. This potential has not been realized for human vaccination, due to the requirement of large antigen doses and toxic (to humans) adjuvants to overcome the induction of oral tolerance and potential degradation of antigens in the stomach. To date, only oral vaccines based on live attenuated organisms have been approved for human use. In this study we describe the use of a lipid-based delivery system/adjuvant, Lipid C, for oral immunization to protect mice against genital tract chlamydial infection. Lipid C is formulated from food-grade purified and fractionated triglycerides. Bacterial shedding following vaginal challenge with Chlamydia muridarum was reduced by 50% in female mice orally immunized with the chlamydial major outer membrane protein (MOMP) formulated in Lipid C, protection equivalent to that seen in animals immunized with MOMP admixed with both cholera toxin (CT) and CpG oligodeoxynucleotides (CpG-ODN). Protection was further enhanced when MOMP, CT and CpG were all combined in the Lipid C matrix. Protection correlated with production of gamma interferon (IFN) by splenic T cells, a serum MOMP-specific IgG response and low but detectable levels of MOMP-specific IgA in vaginal lavage. PMID- 20026450 TI - [Role of the epithelial-mesenchymal transition during tumor progression]. AB - The epithelial-mesenchymal transition (EMT) is a morphogenetic program that converts epithelial into mesenchymal cells during the embryonic development. This mechanism is frequently reactivated during tumor progression and provides cells with motility and invasive capabilities favoring the metastatic dissemination from epithelial tumors. Various EMT-inducing transcription factors, such as the TWIST proteins, were also shown to inhibit oncogene-induced fail-safe programs (senescence and apoptosis), thereby promoting the progression from benign to malignant stages. Altogether, these observations suggest that EMT could play an important role in favoring both tumor development and metastatic dissemination. PMID- 20026451 TI - Web usage data as a means of evaluating public health messaging and outreach. AB - BACKGROUND: The Internet is increasingly utilized by researchers, health care providers, and the public to seek medical information. The Internet also provides a powerful tool for public health messaging. Understanding the needs of the intended audience and how they use websites is critical for website developers to provide better services to the intended users. OBJECTIVE: The aim of the study was to examine the utilization of the chronic fatigue syndrome (CFS) website at the Centers for Disease Control and Prevention (CDC). We evaluated (1) CFS website utilization, (2) outcomes of a CDC CFS public awareness campaign, and (3) user behavior related to public awareness campaign materials and CFS continuing medical education courses. METHODS: To describe and evaluate Web utilization, we collected Web usage data over an 18-month period and extracted page views, visits, referring domains, and geographic locations. We used page views as the primary measure for the CFS awareness outreach effort. We utilized market basket analysis and Markov chain model techniques to describe user behavior related to utilization of campaign materials and continuing medical education courses. RESULTS: The CDC CFS website received 3,647,736 views from more than 50 countries over the 18-month period and was the 33rd most popular CDC website. States with formal CFS programs had higher visiting density, such as Washington, DC; Georgia; and New Jersey. Most visits (71%) were from Web search engines, with 16% from non search-engine sites and 12% from visitors who had bookmarked the site. The public awareness campaign was associated with a sharp increase and subsequent quick drop in Web traffic. Following the campaign, user interest shifted from information targeting consumer basic knowledge to information for health care professionals. The market basket analysis showed that visitors preferred the 60-second radio clip public service announcement over the 30-second one. Markov chain model results revealed that most visitors took the online continuing education courses in sequential order and were less likely to drop out after they reached the Introduction pages of the courses. CONCLUSIONS: The utilization of the CFS website reflects a high level of interest in the illness by visitors to the site. The high utilization shows the website to be an important online resource for people seeking basic information about CFS and for those looking for professional health care and research information. Public health programs should consider analytic methods to further public health by understanding the characteristics of those seeking information and by evaluating the outcomes of public health campaigns. The website was an effective means to provide health information about CFS and serves as an important public health tool for community outreach. PMID- 20026452 TI - Beyond the PhD: putting the right tools in your research toolbox. AB - Postdoctoral training is vital to a successful career for nurse researchers with a biological or biobehavioral focus. Such training provides structured time to devote to gaining substantive knowledge, expanding one's biological-methods repertoire, and writing grants. However, for unknown reasons, relatively few nurses pursue postdoctoral training. A few plausible explanations include a near critical shortage of nursing faculty coupled with an aging population in need of health care, a lack of available mentoring for predoctoral students to pursue postdoctoral training, and the difficulty of navigating the process of finding and choosing the right match for a postdoctoral experience. The purposes of this article are to provide a rationale for choosing postdoctoral training, review common fellowship opportunities, and discuss the process of finding and choosing the right match for postdoctoral training. The authors provide two prospective plans for postdoctoral training and include a plan for staying on track during the postdoctoral experience. PMID- 20026453 TI - Transitions of elders between long-term care and hospitals. AB - Elderly long-term care recipients who require acute hospitalizations must navigate a fragmented system with poor "handoffs," often resulting in negative outcomes. This article makes the case that reducing preventable hospitalizations and improving transitions to and from hospitals will enhance health care quality and outcomes among these elders. Immediate action targeting diffusion of evidence based care is recommended to decrease avoidable rehospitalizations and achieve cost savings. Policy changes are needed to address barriers to high-quality transitional care, including deficits in health professionals' and caregivers' knowledge and resources, regulatory obstacles, and inadequate financial incentives and clinical information systems. PMID- 20026454 TI - Generational differences in registered nurse turnover. AB - The chronic nature of the nursing workforce shortage in the United States is a continuing concern. As the nationwide gap between supply and demand grows, it remains unknown what impact turnover will have on nursing, access to care, and efforts to improve quality and safety of health care. It also remains unclear whether the recent turnover trends among new graduate registered nurses differ from past generational cohorts of new nurses. The aims of this study were to identify the reasons why registered nurses turnover by generational cohort (Veterans, Baby Boomers, and GenXMs) and to compare the length of time nurses were employed in their first five nursing positions by generational cohort. The findings suggest the three generational cohorts displayed similar reasons for leaving nursing positions with relocation, career advancement, and personal/family reasons reported most frequently. Except for the first nursing position, significant generational effects were found in the length of time Veterans, Baby Boomer, and GenXMs stayed employed in their nursing positions. It remains unknown why the GenXMs displayed a significantly shorter length of employment time in their second, third, fourth, and fifth nursing positions. The decline in length of employment time displayed in both the Baby Boomers and GenXMs may be an issue of concern requiring future research. PMID- 20026455 TI - Left ventricular strain and strain rate: characterization of the effect of load in human subjects. AB - AIMS: Left ventricular (LV) strain and strain rate have been proposed as novel indices of systolic function; however, there are limited data about the effect of acute changes on these parameters. METHODS AND RESULTS: Simultaneous Millar micromanometer LV pressure and echocardiographic assessment were performed on 18 patients. Loading was altered sequentially by the administration of glyceryl trinitrate (GTN) and saline fluid loading. Echocardiographic speckle tracking imaging was used to quantify the peak systolic strain (S) and peak systolic strain rate (SR S) and dp/dt max was recorded from the micromanometer data. GTN administration decreased preload (LV end diastolic pressure [LVEDP]: 15.7 vs. 8.4 mmHg, P < 0.001) and afterload (end systolic wall stress: 74 vs. 43 x 10(3)dyn/cm(2), P < 0.001). Administration of fluid increased preload (LVEDP: 11.3 vs. 18.1 mmHg, P < 0.001) and increased wall stress (53 vs. 62 x 10(3)dyn/cm(2), P < 0.003). Administration of GTN resulted in increased circumferential SR S (-1.2 vs. -1.7s(-1), P < 0.01) and longitudinal SR S (-0.9 vs. -1.0 s(-1), P < 0.001). The administration of fluid resulted in decreased circumferential SR S (-1.5 vs. -1.3s(-1), P < 0.01) and longitudinal SR S (-1.0 vs. -0.9s(-1), P < 0.01). As preload and afterload increased, decrease in circumferential SR S (r = 0.63, P < 0.001; r = 0.56, P<0.001) and longitudinal SR S were observed (r = 0.42, P < 0.003; r = 0.49 P < 0.001). CONCLUSION: Circumferential and longitudinal peak strain and systolic strain rate are sensitive to acute changes in load, an important factor that needs to be considered in their application as indices of systolic function. PMID- 20026456 TI - Red blood cell distribution width and 1-year mortality in acute heart failure. AB - AIMS: Red blood cell distribution width (RDW) predicts mortality in chronic heart failure (HF) and stable coronary artery disease. The prognostic value of RDW in more acute settings such as acute HF, and its relative prognostic value compared with more established measures such as N-terminal pro-brain natriuretic peptide (NT-proBNP), remains unknown. METHODS AND RESULTS: In a cohort of 205 patients with acute HF, independent predictors of RDW were identified using linear regression analysis. The association between RDW and 1-year survival in the context of other predictors was assessed using Cox's proportional hazards analysis. Red blood cell distribution width was elevated in 67 (32.7%) patients; RDW was independently associated with haematological variables such as haemoglobin (P < 0.001) as well as the use of loop diuretics (P = 0.006) and beta blockers (P = 0.015) on presentation, but not with nutritional deficiencies, recent transfusion, or inflammatory variables. Log-transformed RDW values independently predicted mortality in multivariable Cox's proportional hazards analysis (hazards ratio, 1.03; 95% confidence interval, 1.00-1.06; P = 0.04); when stratified on the basis of RDW and NT-proBNP status, the combination provided additional prognostic information. CONCLUSION: Red blood cell distribution width is frequently elevated among patients with acute HF and does not appear to be associated with nutritional status, transfusion history, or inflammation. Red blood cell distribution width independently predicts 1-year mortality in acute HF. The value of RDW appears additive to other established prognostic variables such as NT-proBNP. PMID- 20026457 TI - A randomized, double-blind, parallel, comparative study to evaluate the efficacy and safety of ramosetron plus dexamethasone injection for the prevention of acute chemotherapy-induced nausea and vomiting. AB - OBJECTIVE: To evaluate the efficacy of intravenous ramosetron plus dexamethasone for the prevention of acute chemotherapy-induced nausea and vomiting. METHODS: Cancer patients scheduled to receive chemotherapy containing either of the four drugs (cisplatin, doxorubicin, epirubicin or oxaliplatin) were enrolled. They were randomized to receive intravenous ramosetron 0.3 mg plus dexamethasone 20 mg or granisetron 3 mg plus dexamethasone 20 mg 30 min before chemotherapy on day 1. The primary efficacy parameter is complete response rate, which was defined by the proportion of patients without vomiting and no requirement for rescue drugs within 24 h after chemotherapy. RESULTS: A total of 285 patients were enrolled. The primary efficacy analysis included 274 patients. The complete response rate was 77.37% in the ramosetron 0.3 mg plus dexamethasone 20 mg group (137 patients) and 81.75% in the granisetron 3 mg plus dexamethasone 20 mg group (137 patients) with a difference of -4.38% (95% confidence interval: -14.64, 5.89). Therefore, non-inferiority of ramosetron 0.3 mg plus dexamethasone 20 mg to granisetron 3 mg plus dexamethasone 20 mg was demonstrated with non-inferiority margin -15%. For patients treated with cisplatin, non-inferiority of ramosetron 0.3 mg plus dexamethasone 20 mg to granisetron 3 mg plus dexamethasone 20 mg could not be demonstrated. Only a few patients required rescue medications, 7.3% in the ramosetron 0.3 mg plus dexamethasone 20 mg group and 5.1% in the granisetron 3 mg plus dexamethasone 20 mg group (P = 0.44). All 285 patients were included for safety analysis; 36.11% (52/144) and 23.40% (33/141) experienced at least one adverse event within 24 h in the ramosetron 0.3 mg plus dexamethasone 20 mg and granisetron 3 mg plus dexamethasone 20 mg groups, respectively. Four ramosetron related adverse events among 144 patients were observed including two moderate elevation of liver enzymes and one each of mild hiccup and moderate skin rash. CONCLUSIONS: The combination of ramosetron plus dexamethasone was an effective treatment to prevent acute chemotherapy-induced nausea and vomiting. PMID- 20026458 TI - A case with Hodgkin lymphoma and fronto-temporal lobular degeneration (FTLD)-like dementia facilitated by chemotherapy. AB - We report a case of a 39-year-old man with Hodgkin lymphoma who developed depressive symptoms after starting adriamycin, bleomycin, vinblastine and dacarbazine chemotherapy and later exhibited sexual disinhibition in addition to cognitive dysfunction (mainly executive dysfunction). Seven months after the start of adriamycin, bleomycin, vinblastine and dacarbazine chemotherapy, he was finally diagnosed as having fronto-temporal lobular degeneration-like dementia facilitated by adriamycin, bleomycin, vinblastine and dacarbazine chemotherapy. At the time of writing, the patient's condition has persisted for more than 6 months after the discontinuation of adriamycin, bleomycin, vinblastine and dacarbazine chemotherapy, and the changes in brain function brought on by the adriamycin, bleomycin, vinblastine and dacarbazine chemotherapy may now be irreversible. This case points to the importance of being attentive to the appearance of neuropsychiatric symptoms and evaluating brain functions properly when performing anti-cancer chemotherapy. PMID- 20026459 TI - Rationality and emotions. PMID- 20026460 TI - Self-poisoning of the mind. AB - Rational-choice theory tries to explain behaviour on the assumption that individuals optimize. Some forms of irrational behaviour can be explained by assuming that the individual is subject to hedonic, pleasure-seeking mechanisms, such as wishful thinking or adaptive preference formation. In this paper, I draw attention to psychic mechanisms, originating in the individual, which make her worse off. I first consider the ideas of counterwishful thinking and of counteradaptive preference formation and then, drawing heavily on Proust, the self-poisoning of the mind that occurs through the operation of amour-propre. PMID- 20026461 TI - Self-deception as self-signalling: a model and experimental evidence. AB - Self-deception has long been the subject of speculation and controversy in psychology, evolutionary biology and philosophy. According to an influential 'deflationary' view, the concept is an over-interpretation of what is in reality an instance of motivationally biased judgement. The opposite view takes the interpersonal deception analogy seriously, and holds that some part of the self actively manipulates information so as to mislead the other part. Building on an earlier self-signalling model of Bodner and Prelec, we present a game-theoretic model of self-deception. We propose that two distinct mechanisms collaborate to produce overt expressions of belief: a mechanism responsible for action selection (including verbal statements) and an interpretive mechanism that draws inferences from actions and generates emotional responses consistent with the inferences. The model distinguishes between two modes of self-deception, depending on whether the self-deceived individual regards his own statements as fully credible. The paper concludes with a new experimental study showing that self-deceptive judgements can be reliably and repeatedly elicited with financial incentives in a categorization task, and that the degree of self-deception varies with incentives. The study also finds evidence of the two forms of self-deception. The psychological benefits of self-deception, as measured by confidence, peak at moderate levels. PMID- 20026462 TI - Counterfactual thinking and emotions: regret and envy learning. AB - Emotions like regret and envy share a common origin: they are motivated by the counterfactual thinking of what would have happened had we made a different choice. When we contemplate the outcome of a choice we made, we may use the information on the outcome of a choice we did not make. Regret is the purely private comparison between two choices that we could have taken, envy adds to this the information on outcome of choices of others. However, envy has a distinct social component, in that it adds the change in the social ranking that follows a difference in the outcomes. We study the theoretical foundation and the experimental test of this view. PMID- 20026463 TI - Regret and the rationality of choices. AB - Regret helps to optimize decision behaviour. It can be defined as a rational emotion. Several recent neurobiological studies have confirmed the interface between emotion and cognition at which regret is located and documented its role in decision behaviour. These data give credibility to the incorporation of regret in decision theory that had been proposed by economists in the 1980s. However, finer distinctions are required in order to get a better grasp of how regret and behaviour influence each other. Regret can be defined as a predictive error signal but this signal does not necessarily transpose into a decision-weight influencing behaviour. Clinical studies on several types of patients show that the processing of an error signal and its influence on subsequent behaviour can be dissociated. We propose a general understanding of how regret and decision making are connected in terms of regret being modulated by rational antecedents of choice. Regret and the modification of behaviour on its basis will depend on the criteria of rationality involved in decision-making. We indicate current and prospective lines of research in order to refine our views on how regret contributes to optimal decision-making. PMID- 20026464 TI - Rational choice, neuroeconomy and mixed emotions. AB - Experimental psychology has shown differences between predictions of theory of decision and human choices. Emotions like regret can partly explain these differences. Neuroimagery used in combination with behavioural economics (neuroeconomics) has been used in order to try to disentangle the different emotional and rational factors (regret, rejoicing, reward, costs, uncertainty, trade-off between positive and negative aspects of different options). Emotions then appear as much more complex and mixed affective states than usually assumed. Not only might we feel a positive affect in punishing unfair partners, but mixed emotions can, for example, combine transmutation of previous anxiety into relief and elation by comparison with another less exciting option (elating relief). At the level of complexity of these mixed emotions--which we formally represent by comparisons between 'unexpected utilities' and expected ones--the main biases that Kahnemann and Tversky have shown can be explained. In spite of the complexity of these mixed emotions, some of these hypotheses might be partially tested by brain imagery. PMID- 20026465 TI - Changing time and emotions. AB - In this paper, we consider that our experience of time (to come) depends on the emotions we feel when we imagine future pleasant or unpleasant events. A positive emotion such as relief or joy associated with a pleasant event that will happen in the future induces impatience. Impatience, in our context, implies that the experience of time up to the forthcoming event expands. A negative emotion such as grief or frustration associated with an unpleasant event that will happen in the future triggers anxiety. This will give the experience of time contraction. Time, therefore, is not exogeneously given to the individual and emotions, which link together events or situations, are a constitutive ingredient of the experience of time. Our theory can explain experimental evidence that people tend to prefer to perform painful actions earlier than pleasurable ones, contrary to the predictions yielded by the standard exponential discounting framework. PMID- 20026466 TI - Herding, social influence and economic decision-making: socio-psychological and neuroscientific analyses. AB - Typically, modern economics has steered away from the analysis of sociological and psychological factors and has focused on narrow behavioural assumptions in which expectations are formed on the basis of mathematical algorithms. Blending together ideas from the social and behavioural sciences, this paper argues that the behavioural approach adopted in most economic analysis, in its neglect of sociological and psychological forces and its simplistically dichotomous categorization of behaviour as either rational or not rational, is too narrow and stark. Behaviour may reflect an interaction of cognitive and emotional factors and this can be captured more effectively using an approach that focuses on the interplay of different decision-making systems. In understanding the mechanisms affecting economic and financial decision-making, an interdisciplinary approach is needed which incorporates ideas from a range of disciplines including sociology, economic psychology, evolutionary biology and neuroeconomics. PMID- 20026467 TI - Embodied economics: how bodily information shapes the social coordination dynamics of decision-making. AB - To date, experiments in economics are restricted to situations in which individuals are not influenced by the physical presence of other people. In such contexts, interactions remain at an abstract level, agents guessing what another person is thinking or is about to decide based on money exchange. Physical presence and bodily signals are therefore left out of the picture. However, in real life, social interactions (involving economic decisions or not) are not solely determined by a person's inference about someone else's state-of-mind. In this essay, we argue for embodied economics: an approach to neuroeconomics that takes into account how information provided by the entire body and its coordination dynamics influences the way we make economic decisions. Considering the role of embodiment in economics--movements, posture, sensitivity to mimicry and every kind of information the body conveys--makes sense. This is what we claim in this essay which, to some extent, constitutes a plea to consider bodily interactions between agents in social (neuro)economics. PMID- 20026468 TI - Selfish or selfless? The role of empathy in economics. AB - Empathy is a longstanding issue in economics, especially for welfare economics, but one which has faded from the scene in recent years. However, with the rise of neuroeconomics, there is now a renewed interest in this subject. Some economists have even gone so far as to suggest that neuroscientific experiments reveal heterogeneous empathy levels across individuals. If this were the case, this would be in line with economists' usual assumption of stable and given preferences and would greatly facilitate the study of prosocial behaviour with which empathy is often associated. After reviewing some neuroscientific psychological and neuroeconomic evidence on empathy, we will, however, criticize the notion of a given empathy distribution in the population by referring to recent experiments on a public goods game that suggest that, on the contrary, the degree of empathy that individuals exhibit is very much dependent on context and social interaction. PMID- 20026471 TI - REACH--how is it going? PMID- 20026469 TI - Decision-making during gambling: an integration of cognitive and psychobiological approaches. AB - Gambling is a widespread form of entertainment that may afford unique insights into the interaction between cognition and emotion in human decision-making. It is also a behaviour that can become harmful, and potentially addictive, in a minority of individuals. This article considers the status of two dominant approaches to gambling behaviour. The cognitive approach has identified a number of erroneous beliefs held by gamblers, which cause them to over-estimate their chances of winning. The psychobiological approach has examined case-control differences between groups of pathological gamblers and healthy controls, and has identified dysregulation of brain areas linked to reward and emotion, including the ventromedial prefrontal cortex (vmPFC) and striatum, as well as alterations in dopamine neurotransmission. In integrating these two approaches, recent data are discussed that reveal anomalous recruitment of the brain reward system (including the vmPFC and ventral striatum) during two common cognitive distortions in gambling games: the near-miss effect and the effect of personal control. In games of chance, near-misses and the presence of control have no objective influence on the likelihood of winning. These manipulations appear to harness a reward system that evolved to learn skill-oriented behaviours, and by modulating activity in this system, these cognitive distortions may promote continued, and potentially excessive, gambling. PMID- 20026470 TI - From molecule to market: steroid hormones and financial risk-taking. AB - Little is known about the role of the endocrine system in financial decision making. Here, we survey research on steroid hormones and their cognitive effects, and examine potential links to trader performance in the financial markets. Preliminary findings suggest that cortisol codes for risk and testosterone for reward. A key finding of this endocrine research is the different cognitive effects of acute versus chronic exposure to hormones: acutely elevated steroids may optimize performance on a range of tasks; but chronically elevated steroids may promote irrational risk-reward choices. We present a hypothesis suggesting that the irrational exuberance and pessimism observed during market bubbles and crashes may be mediated by steroid hormones. If hormones can exaggerate market moves, then perhaps the age and sex composition among traders and asset managers may affect the level of instability witnessed in the financial markets. PMID- 20026472 TI - Toxicity testing in the 21st century: using the new toxicity testing paradigm to create a taxonomy of adverse effects. AB - Distinguishing adaptive from adverse responses is fundamental to understanding toxicity and to implementing regulatory guidelines that are protective of human health. However, what we consider to be an adverse effect may change over time as the cultural acceptance of risk alters and new knowledge and insight accumulate. The fact that the identification of an adverse effect is subject to change is obvious, necessary, and uncomfortable. In this commentary, a framework for defining adverse effects is proposed for the emerging paradigm of toxicity testing in the 21st century-a paradigm that focuses on human cells, in vitro approaches, toxicity pathways, and high-throughput techniques. The traditional meaning of an adverse effect as a change at the organismal level is not compatible with this new system of toxicity testing. Instead, based on the experience of accident investigators, we propose that a Toxicological Factors Analysis and Classification System will use the database resulting from the high throughput toxicity testing of the future to develop a Taxonomy of Adverse Effects. Similar to an accident, predisposing "latent failures" identified within categories of the toxicant response database will be associated with the "active failure" of an adverse effect. PMID- 20026473 TI - Integration of in vivo genotoxicity and short-term carcinogenicity assays using F344 gpt delta transgenic rats: in vivo mutagenicity of 2,4-diaminotoluene and 2,6-diaminotoluene structural isomers. AB - An important trend in current toxicology is the replacement, reduction, and refinement of the use of experimental animals (the 3R principle). We propose a model in which in vivo genotoxicity and short-term carcinogenicity assays are integrated with F344 gpt delta transgenic rats. Using this model, the genotoxicity of chemicals can be identified in target organs using a shuttle vector lambda EG10 that carries reporter genes for mutations; short-term carcinogenicity is determined by the formation of glutathione S-transferase placenta form (GST-P) foci in the liver. To begin validating this system, we examined the genotoxicity and hepatotoxicity of structural isomers of 2,4 diaminotoluene (2,4-DAT) and 2,6-diaminotoluene (2,6-DAT). Although both compounds are genotoxic in the Ames/Salmonella assay, only 2,4-DAT induces tumors in rat livers. Male F344 gpt delta rats were fed diet containing 2,4-DAT at doses of 125, 250, or 500 ppm for 13 weeks or 2,6-DAT at a dose of 500 ppm for the same period. The mutation frequencies of base substitutions, mainly at G:C base pairs, were significantly increased in the livers of 2,4-DAT-treated rats at all three doses. In contrast, virtually no induction of genotoxicity was identified in the kidneys of 2,4-DAT-treated rats or in the livers of 2,6-DAT-treated rats. GST-P positive foci were detected in the livers of rats treated with 2,4-DAT at a dose of 500 ppm but not in those treated with 2,6-DAT. Integrated genotoxicity and short-term carcinogenicity assays may be useful for early identifying genotoxic and nongenotoxic carcinogens in a reduced number of experimental animals. PMID- 20026474 TI - Metabolic turnover analysis by a combination of in vivo 13C-labelling from 13CO2 and metabolic profiling with CE-MS/MS reveals rate-limiting steps of the C3 photosynthetic pathway in Nicotiana tabacum leaves. AB - Understanding of the control of metabolic pathways in plants requires direct measurement of the metabolic turnover rate. Sugar phosphate metabolism, including the Calvin cycle, is the primary pathway in C(3) photosynthesis, the dynamic status of which has not been assessed quantitatively in the leaves of higher plants. Since the flux of photosynthetic carbon metabolism is affected by the CO(2) fixation rate in leaves, a novel in vivo (13)C-labelling system was developed with (13)CO(2) for the kinetic determination of metabolic turnover that was the time-course of the (13)C-labelling ratio in each metabolite. The system is equipped with a gas-exchange chamber that enables real-time monitoring of the CO(2) fixation rate and a freeze-clamp that excises a labelled leaf concurrently with quenching the metabolic reactions by liquid nitrogen within the photosynthesis chamber. Kinetic measurements were performed by detecting mass isotopomer abundance with capillary electrophoresis-tandem mass spectrometry. The multiple reaction monitoring method was optimized for the determination of each compound for sensitive detection because the amount of some sugar phosphates in plant cells is extremely small. Our analytical system enabled the in vivo turnover of sugar phosphates to be monitored in fresh tobacco (Nicotiana tabacum) leaves, which revealed that the turnover rate of glucose-1-phosphate (G1P) was significantly lower than that of other sugar phosphates, including glucose-6 phosphate (G6P). The pool size of G1P is 12 times lower than that of G6P. These results indicate that the conversion of G6P to G1P is one of the rate-limiting steps in the sugar phosphate pathway. PMID- 20026475 TI - Communication skills training in oncology: a position paper based on a consensus meeting among European experts in 2009. AB - BACKGROUND: Communication in cancer care has become a major topic of interest. Since there is evidence that ineffective communication affects both patients and oncology clinicians (physicians and nurses), so-called communication skills trainings (CSTs) have been developed over the last decade. While these trainings have been demonstrated to be effective, there is an important heterogeneity with regard to implementation and with regard to evidence of different aspects of CST. METHODS: In order to review and discuss the scientific literature on CST in oncology and to formulate recommendations, the Swiss Cancer League has organised a consensus meeting with European opinion leaders and experts in the field of CST, as well as oncology clinicians, representatives of oncology societies and patient organisations. On the basis of a systematic review and a meta-analysis, recommendations have been developed and agreed upon. RESULTS: Recommendations address (i) the setting, objectives and participants of CST, (ii) its content and pedagogic tools, (iii) organisational aspects, (iv) outcome and (v) future directions and research. CONCLUSION: This consensus meeting, on the basis of European expert opinions and a systematic review and meta-analysis, defines key elements for the current provision and future development and evaluation of CST in oncology. PMID- 20026476 TI - A quantitative proteomics analysis of subcellular proteome localization and changes induced by DNA damage. AB - A major challenge in cell biology is to identify the subcellular distribution of proteins within cells and to characterize how protein localization changes under different cell growth conditions and in response to stress and other external signals. Protein localization is usually determined either by microscopy or by using cell fractionation combined with protein blotting techniques. Both these approaches are intrinsically low throughput and limited to the analysis of known components. Here we use mass spectrometry-based proteomics to provide an unbiased, quantitative, and high throughput approach for measuring the subcellular distribution of the proteome, termed "spatial proteomics." The spatial proteomics method analyzes a whole cell extract created by recombining differentially labeled subcellular fractions derived from cells in which proteins have been mass-labeled with heavy isotopes. This was used here to measure the relative distribution between cytoplasm, nucleus, and nucleolus of over 2,000 proteins in HCT116 cells. The data show that, at steady state, the proteome is predominantly partitioned into specific subcellular locations with only a minor subset of proteins equally distributed between two or more compartments. Spatial proteomics also facilitates a proteome-wide comparison of changes in protein localization in response to a wide range of physiological and experimental perturbations, shown here by characterizing dynamic changes in protein localization elicited during the cellular response to DNA damage following treatment of HCT116 cells with etoposide. DNA damage was found to cause dissociation of the proteasome from inhibitory proteins and assembly chaperones in the cytoplasm and relocation to associate with proteasome activators in the nucleus. PMID- 20026477 TI - Predicting Arabidopsis freezing tolerance and heterosis in freezing tolerance from metabolite composition. AB - Heterosis, or hybrid vigor, is one of the most important tools in plant breeding and has previously been demonstrated for plant freezing tolerance. Freezing tolerance is an important trait because it can limit the geographical distribution of plants and their agricultural yield. Plants from temperate climates increase in freezing tolerance during exposure to low, non-freezing temperatures in a process termed 'cold acclimation'. Metabolite profiling has indicated a major reprogramming of plant metabolism in the cold, but it has remained unclear in previous studies which of these changes are related to freezing tolerance. In the present study, we have used metabolic profiling to discover combinations of metabolites that predict freezing tolerance and its heterosis in Arabidopsis thaliana. We identified compatible solutes and, in particular, the pathway leading to raffinose as crucial statistical predictors for freezing tolerance and its heterosis, while some TCA cycle intermediates contribute only to predicting the heterotic phenotype. This indicates coordinate links between heterosis and metabolic pathways, suggesting that a limited number of regulatory genes may determine the extent of heterosis in this complex trait. In addition, several unidentified metabolites strongly contributed to the prediction of both freezing tolerance and its heterosis and we present an exemplary analysis of one of these, identifying it as a hexose conjugate. PMID- 20026478 TI - Evolution of Caenorhabditis mitochondrial genome pseudogenes and Caenorhabditis briggsae natural isolates. AB - Although most metazoan mitochondrial genomes are highly streamlined and encode little noncoding DNA outside of the "AT" region, the accumulation of mitochondrial pseudogenes and other types of noncoding DNA has been observed in a growing number of animal groups. The nematode species Caenorhabditis briggsae harbors two mitochondrial DNA (mtDNA) pseudogenes, named Psinad5-1 and Psinad5-2, presumably derived from the nad5 protein-coding gene. Here, we provide an in depth analysis of mtDNA pseudogene evolution in C. briggsae natural isolates and related Caenorhabditis species. Mapping the observed presence and absence of the pseudogenes onto phylogenies suggests that Psinad5-1 originated in the ancestor to C. briggsae and its recently discovered outcrossing relative species Caenorhabditis sp. 5 and Caenorhabditis sp. 9. However, Psinad5-1 was not detected in Caenorhabditis sp. 9 natural isolates, suggesting a lineage-specific loss of this pseudogene in this species. Our results corroborated the previous finding that Psinad5-2 originated within C. briggsae. The observed pattern of mitochondrial pseudogene gain and loss in Caenorhabditis was inconsistent with predictions of the tandem duplication-random loss model of mitochondrial genome evolution and suggests that intralineage recombination-like mechanisms might play a major role in Caenorhabditis mtDNA evolution. Natural variation was analyzed at the pseudogenes and flanking mtDNA sequences in 141 geographically diverse C. briggsae natural isolates. Although phylogenetic analysis placed the majority of isolates into the three previously established major intraspecific clades of C. briggsae, two new and unexpected haplotypes fell outside of these conventional groupings. Psinad5-2 copy number variation was observed among C. briggsae isolates collected from the same geographic site. Patterns of nucleotide diversity were analyzed in Psinad5-1 and Psinad5-2, and confidence intervals were found to overlap values from synonymous sites in protein-coding genes, consistent with neutral expectations. Our findings provide new insights into the mode and tempo of mitochondrial genome and pseudogene evolution both within and between Caenorhabditis nematode species. PMID- 20026479 TI - Rapid proliferation of repetitive palindromic elements in mtDNA of the endemic Baikalian sponge Lubomirskia baicalensis. AB - Animal mitochondrial DNA (mtDNA) is a remarkably compact molecule largely because of the scarcity of noncoding "selfish" DNA. Recently, however, we found that mitochondrial genomes of several phylogenetically diverse species of demosponges contain small repetitive palindromic sequences, interspersed within intergenic regions and fused in protein and ribosomal RNA genes. Here, I report and analyze the proliferation of such elements in the mitochondrial genome of the endemic sponge of Lake Baikal Lubomirskia baicalensis. Because Baikal sponges are closely related to the circumglobally distributed freshwater sponge Ephydatia muelleri with which they shared a common ancestor approximately 3-10 Ma, both the rate of single nucleotide substitutions and the rate of palindromic repeat insertions can be calculated in this system. I found the rate of nucleotide substitutions in mtDNA of freshwater sponges to be extremely low (0.5-1.6 x 10(-9) per site per year), more similar to that in plants than bilaterian animals. By contrast, the per/nucleotide rate of insertions of repetitive elements is at least four times higher. This rapid rate of proliferation combined with the broad phylogenetic distribution of hairpin elements can make them a defining force in the evolution of mitochondrial genomes of demosponges. PMID- 20026480 TI - The increase in the number of subunits in eukaryotic RNA polymerase III relative to RNA polymerase II is due to the permanent recruitment of general transcription factors. AB - The number of subunits of RNA polymerases (RNAPs) increases during evolution from 5 in eubacteria to 12 in archaea. In eukaryotes, which have at least three RNAPs, the number of subunits has expanded from 12 in RNA polymerase II (RNAPII) to 14 in RNA polymerase I (RNAPI) and to 17 in RNA polymerase III (RNAPIII). It was recently demonstrated that the two additional subunits found in RNAPI relative to RNAPII are homologous to TFIIF, a dimeric general transcription factor of RNAPII. Here, we extend this finding by demonstrating that four of the five RNAPIII specific subunits are also homologous to transcription factors of RNAPII. We use the available evidence to propose an evolutionary history of the eukaryotic RNAPs and argue that the increases in the number of subunits that occurred in RNAPs I and III are due to the permanent recruitment of preexisting transcription factors. PMID- 20026481 TI - Timing of replication is a determinant of neutral substitution rates but does not explain slow Y chromosome evolution in rodents. AB - Mutation rates, assayed as substitution rates of putatively neutral sites, are highly variable around mammalian genomes: There is heterogeneity between genes, between autosomes, and between X, Y, and autosomes. The differences between X, Y, and autosomes are typically assumed to reflect the greater number of cell divisions in the male germ-line. Such an effect can neither account for within autosome differences nor does it predict the differences between X, Y, and autosome observed in rodents. It has recently been proposed that in primates, the time during S-phase when a gene is replicated is an important determinant of neutral rates of evolution. Here we ask 1) whether we can replicate this result in rodents, 2) whether different autosomes replicate on average at different times, and 3) whether this might explain differences in their substitution rates. Finally we ask 4) whether X, Y, and autosome replicate at different times and 5) whether any difference might explain why the number of replication events alone cannot explain their substitution rates. We find that, as in primates, autosomal intronic rates of evolution increase significantly during S-phase. Different autosomes do have different average replication times, and together with rearrangement, this is a significant predictor of between-autosome differences in substitution rate. Although we find that autosomal, X-, and Y-linked genes replicate at different times, it is paradoxical that the Y-linked genes replicate latest, and replicate more often, but are not especially fast evolving. These results support the hypothesis that replication timing is an important source of substitution rate heterogeneity. PMID- 20026482 TI - Evaluating the relationship between evolutionary divergence and phylogenetic accuracy in AFLP data sets. AB - Using in silico amplified fragment length polymorphism (AFLP) fingerprints, we explore the relationship between sequence similarity and phylogeny accuracy to test when, in terms of genetic divergence, the quality of AFLP data becomes too low to be informative for a reliable phylogenetic reconstruction. We generated DNA sequences with known phylogenies using balanced and unbalanced trees with recent, uniform and ancient radiations, and average branch lengths (from the most internal node to the tip) ranging from 0.02 to 0.4 substitutions per site. The resulting sequences were used to emulate the AFLP procedure. Trees were estimated by maximum parsimony (MP), neighbor-joining (NJ), and minimum evolution (ME) methods from both DNA sequences and virtual AFLP fingerprints. The estimated trees were compared with the reference trees using a score that measures overall differences in both topology and relative branch length. As expected, the accuracy of AFLP-based phylogenies decreased dramatically in the more divergent data sets. Above a divergence of approximately 0.05, AFLP-based phylogenies were largely inaccurate irrespective of the distinct topology, radiation model, or phylogenetic method used. This value represents an upper bound of expected tree accuracy for data sets with a simple divergence history; AFLP data sets with a similar divergence but with unbalanced topologies and short ancestral branches produced much less accurate trees. The lack of homology of AFLP bands quickly increases with divergence and reaches its maximum value (100%) at a divergence of only 0.4. Low guanine-cytosine (GC) contents increase the number of nonhomologous bands in AFLP data sets and lead to less reliable trees. However, the effect of the lack of band homology on tree accuracy is surprisingly small relative to the negative impact due to the low information content of AFLP characters. Tree building methods based on genetic distance displayed similar trends and outperformed parsimony at low but not at high divergences. However, the impact of using alternative phylogenetic methods on tree accuracy was generally small relative to the uncertainty arising from factors such as divergence, nonhomology of bands, or the low information content of AFLP characters. Nevertheless, our data suggest that under certain circumstances, AFLPs may be suitable to reconstruct deeper phylogenies than usually accepted. PMID- 20026483 TI - Functional diversity of signaling pathways through G protein-coupled receptor heterodimerization with a species-specific orphan receptor subtype. AB - Gonadotropin-releasing hormones (GnRHs) play pivotal roles in control of reproduction via a hypothalamic-pituitary-periphery endocrine system and nervous systems of not only vertebrates but also invertebrates. GnRHs trigger several signal transduction cascades via GnRH receptors (GnRHRs), members of the G protein-coupled receptor (GPCR) family. Recently, six GnRHs (tunicate GnRH [tGnRH]-3 to tGnRH-8) and four GnRHRs (Ciona intestinalis [Ci]-GnRHR1 to GnRHR 4), including a species-specific paralog, Ci-GnRHR4 (R4) regarded as an orphan receptor or nonfunctional receptor, were identified in the protochordate, C. intestinalis, which lacks the hypothalamic-pituitary system. Here, we show novel functional modulation of GnRH signaling pathways via GPCR heterodimerization. Immunohistochemical analysis showed colocalization of R1 and R4 in test cells of the ascidian ovary. The native R1-R4 heterodimerization was detected in the Ciona ovary by coimmunoprecipitation analysis. The heterodimerization in HEK293 cells cotransfected with R1 and R4 was also observed by coimmunoprecipitation and fluorescent energy transfer analyses. Binding assay revealed that R4 had no affinity for tGnRHs, and the heterodimerization did not alter the binding affinity of R1 to the ligands. The R1-R4 elicited 10-fold more potent Ca2+ mobilization than R1 exclusively by tGnRH-6, although R1-mediated cyclic AMP production was not affected by any of tGnRHs via the R1-R4 heterodimer. Moreover, the R1-R4 heterodimer potentiated translocation of both Ca2+-dependent protein kinase C-alpha (PKCalpha) by tGnRH-6 and Ca2+-independent PKCzeta by tGnRH-5 and tGnRH-6, eventually leading to the upregulation of extracellular signal-regulated kinase (ERK) phosphorylation compared with R1 alone. These results provide evidence that the species-specific GnRHR orphan paralog, R4, serves as an endogenous modulator for the fine-tuning of activation of PKC subtype-selective signal transduction via heterodimerization with R1 and that the species-specific GPCR heterodimerization, in concert with multiplication of tGnRHs and Ci-GnRHRs, participates in functional evolution of neuropeptidergic GnRH signaling pathways highly conserved throughout the animal kingdom. PMID- 20026484 TI - Evolutionary expansion and specialization of the PDZ domains. AB - PDZ domains are protein-protein interaction modules widely used to assemble membranous signaling complexes including those found in the neuronal synapse. PDZ containing genes encoded in metazoan genomes vastly outnumber those in prokaryotes, plants, and fungi. By comparing 40 proteomes to track the evolutionary history of the PDZ domain, we observed that the variety of associations between PDZ and other domains expands greatly along the stem leading to metazoans and choanoflagellates. We asked whether the expansion of PDZ domains was due to random or specific sequence changes. Studying the sequence signatures of 58 PDZ lineages that are common to bilaterian animals, we showed that six common amino acid residues are able to classify 96% of PDZ domains to their correct evolutionary lineage. In PDZ domain-ligand cocrystals, four of these "classifying positions" lie in direct contact with the -1 and -3 residues of the ligand. This suggests coevolution of the more flexible regions of the binding interaction as a central mechanism of specialization inherent within the PDZ domain. To identify these positions, we devised two independent algorithms--a metric termed within-clade entropy (WCE) and an average mutual information (AvgMI) score--that both reached similar results. Extending these tools to the choanoflagellate, Monosiga brevicollis, we compared its PDZ domains with their putative metazoan orthologs. Interestingly, the M. brevicollis genes lack conservation at the classifying positions suggesting dissociation between domain organization in multidomain proteins and specific changes within the PDZ domain. PMID- 20026485 TI - Dominance of vision over proprioception on motor programming: evidence from ERP. AB - The brain determines positions and movements of body parts from inputs arising at least from vision and proprioception. Using the brain event-related potential called the lateralized readiness potential, which reflects motor cortical activity during motor programming, we showed in a motor task that viewing one hand in a sagittal mirror-giving the impression to see the opposite hand generated activity in the motor cortex of the seen hand (i.e., of the nonmoving hand hidden behind the mirror). The visual influence on cortical motor region occurred even when the proprioceptive input related to the real opposite effector was not aligned on the visual feedback of the hand given by the mirror. This dominance vision over proprioception was greatly reduced when the task was executed in the dark with hand position represented by small lights fixed on the moving hand, with no motor activity being recorded in the cortical area of the inactive hand. These results give new insights into how the brain weights and integrates visual and proprioceptive information in motor control. PMID- 20026487 TI - Necrotizing fasciitis following drainage of Streptococcus milleri empyema. AB - Streptococcus milleri (SM) is a heterogeneous group of Streptococci, which is a recognized cause of purulent infections of the mediastinal and pleural spaces. These infections are notoriously resistant and require aggressive surgical management. We present our experience with a 60-year-old patient, who developed necrotizing fasciitis of the chest wall after initial bedside drainage of a SM empyema. He required extensive debridement with significant soft tissue loss and subsequent latissimus dorsi flap reconstruction to cover the defect. PMID- 20026486 TI - Correlated low-frequency BOLD fluctuations in the resting human brain are modulated by recent experience in category-preferential visual regions. AB - The resting brain is associated with significant intrinsic activity fluctuations, such as the correlated low-frequency (LF) blood oxygen level-dependent (BOLD) fluctuations measured by functional magnetic resonance imaging. Despite a recent expansion of studies investigating resting-state LF-BOLD correlations, their nature and function are poorly understood. A major constraint on LF-BOLD correlations appears to be stable properties of anatomic connectivity. There is also evidence that coupling can be modulated by recent or ongoing task performance, suggesting that certain components of correlated dynamics are malleable on short timescales. Here, we compared activity during extended periods of rest following performance of 2 distinct cognitive tasks using different categories of visual stimuli-faces and complex scenes. Prolonged exposure to these distinct categories of visual information caused frontal networks to couple differentially with posterior category-preferential visual regions during subsequent periods of rest. In addition, we report preliminary evidence suggesting that conditions exist in which the degree of modulation of LF-BOLD correlations predicts subsequent memory. The finding that resting-state LF-BOLD correlations are modulated by recent experience in functionally specific brain regions engaged during prior task performance clarifies their role as a dynamic phenomenon which may be involved in mnemonic processes. PMID- 20026488 TI - Clinical performance and biocompatibility of hyaluronan-based heparin-bonded extracorporeal circuits in different risk cohorts. AB - This prospective randomized study compares novel hyaluronan-based heparin-bonded circuits vs. uncoated controls across EuroSCORE patient risk strata including biomaterial evaluation. Over a two-year period, 90 patients undergoing coronary artery bypass grafting were prospectively randomized to one of the two perfusion protocols: Group 1 was treated with hyaluronan-based heparin-bonded preconnected circuits (Vision HFO-GBS, Gish, CA, USA) and Group 2 with identical uncoated controls. Each group was composed of three subgroups (n=15) with respect to preoperative evaluation of low (EuroSCORE 0-2), medium (3-5) and high (6+) risk patients. Blood samples were collected after induction (T1) and heparinization (T2), 15 min after cardiopulmonary bypass start (T3), before cessation of CPB (T4), 15 min after reversal (T5), and the first postoperative day (T6). In high risk patients, platelet counts demonstrated significant preservation at T4, T5 and leukocyte counts were lower at T5 in hyaluronan group (P300 seconds (40 vs 20 long events). Longer averaging time tended to underestimate desaturation events of greater severity when SPo(2) was <70% (P = .01). Averaging time did not affect the time spent within various target SPo(2) ranges (P > .05). CONCLUSIONS: These preliminary results suggest that use of longer averaging time reduces the detection of brief periodic desaturation events and of greater severity. It may also interpret a cluster of shorter events as a single, prolonged episode and, thus, potentially overestimate the frequency of long events. The significance of these findings lies in the potential impact on neurodevelopmental outcomes and growth, which will need additional study. PMID- 20026497 TI - Neonatal screening for treatable and untreatable disorders: prospective parents' opinions. AB - OBJECTIVE: In the Netherlands, in 2007, the national newborn screening program was expanded from 3 to 17 disorders that met the World Health Organization's Wilson and Jungner screening criteria, especially regarding treatability. The decision of whether to add diseases to the program is generally based on experts' advice, whereas the opinion of those whom it concerns--prospective parents- remains unknown. In this study, we investigated the opinion of prospective parents concerning newborn screening for disorders that are incurable yet treatable to some extent or even untreatable. METHODS: A structured questionnaire that consisted of 3 parts in which similar questions were posed about treatable, less treatable, and untreatable childhood-onset disorders was posted on the Web site of a national pregnancy fair. RESULTS: A total of 1631 prospective parents filled out the questionnaire, 259 of whom were excluded. In contrast to current policy, respondents showed a positive attitude toward inclusion of less treatable (88%) or untreatable childhood-onset disorders (73%) within the national newborn screening program. Respondents who already had children at the time of completing the questionnaire were even more in favor of screening for especially untreatable disorders. The most important reason mentioned was to prevent a long diagnostic quest. Obtaining information to enable reproductive choices in future pregnancies was hardly mentioned. CONCLUSIONS: Prospective parents in the Dutch population seem interested in newborn screening for untreatable childhood-onset disorders; therefore, we argue that additional debate of pros and cons is needed among policy makers, health care professionals, and consumers. PMID- 20026498 TI - Brain tumor signs and symptoms: analysis of primary health care records from the UKCCS. AB - OBJECTIVE: To compare the frequency of brain tumor signs and symptoms in children with and without brain tumors. METHODS: This was a UK population-based retrospective analysis of primary care records. Participants were 195 children (1 14 years) newly diagnosed with brain tumors and 285 controls matched by age, gender, and region. Comparisons included total number of prediagnosis consultations, number with >or=1 symptom suggestive of a brain tumor, total number of symptoms, number of different symptoms, and number of visits with specific combinations of symptoms. RESULTS: On average, cases consulted more often than controls between birth and diagnosis/pseudodiagnosis with brain tumor signs and symptoms. Their consultation rate with >or=1 suggestive symptom escalated in the 2 years before diagnosis. Symptom prevalence was higher among cases than controls, a relative difference of 3.29 times as many consultations with >or=1 suggestive symptom (95% confidence interval [CI]: 2.82-3.83) and 7.01 as many with more than 1 (95% CI: 5.38-9.13). In each 6-month period in the 4 years before diagnosis, cases had at least twice as many consultations with >or=1 suggestive symptom (20.81 times as many in the 6 months before diagnosis [95% CI: 14.29-30.30]) and 2-3 times more records of suggestive symptoms (28.35 times more in the 6 months before diagnosis [95% CI: 19.05-42.19]). Symptoms rarely or not observed among control children included head tilt, odd head movements, odd posture, back or neck stiffness, and unsteadiness without obvious cause. CONCLUSION Key to identifying the 1 child among many who merits prompt investigation is recognition of unusual symptoms, or specific symptom patterns. PMID- 20026500 TI - Streptococcus pneumoniae--associated hemolytic uremic syndrome: classification and the emergence of serotype 19A. AB - Streptococcus pneumoniae-associated hemolytic uremic syndrome (HUS) is an underrecognized condition that mainly occurs in young children. Early diagnosis is important because of the potential to improve morbidity and mortality rates. The purposes of this report are to review the clinical and laboratory features of 14 patients with pneumococcal HUS and present a modified classification to capture cases that may not have been documented with a diagnosis of pneumococcal HUS. We thereby provide a rationale for including patients with concurrent disseminated intravascular coagulopathy and/or those whose culture results were negative, and we highlight the emergence of serotype 19A subsequent to the introduction of 7-valent pneumococcal protein conjugate vaccine (Prevnar). This is the largest series of such subjects (to our knowledge) from a single center. Sixty-four percent of the patients recovered without any long-term sequelae. Three patients developed chronic kidney disease, 1 developed end-stage renal failure, and 1 died in the acute phase. The greatest risk factor for the development of chronic kidney disease is the need for acute dialysis for >20 days, and death in the acute phase is rare unless meningitis is the primary infection. PMID- 20026499 TI - Prenatal low-dose aspirin and neurobehavioral outcomes of children born very preterm. AB - OBJECTIVE: Low-dose aspirin (LDA) given during pregnancy may alter brain development in very preterm infants. We report the short- and long-term outcomes of very preterm infants according to LDA treatment. PATIENTS AND METHODS: Data were from the Etude Epidemiologique des Petites Ages Gestationnels (EPIPAGE) cohort study, which included all infants born before 33 weeks of gestation in 9 French regions in 1997. This study was restricted to 656 children who were born to 584 women with an obstetric history of placental vascular disease or with chronic hypertension or renal or autoimmune diseases. The main outcome measures were mortality, cerebral lesions, and outcome at 5 years of age, which were measured by a diagnosis of cerebral palsy; behavioral difficulties, which were assessed with the Strength and Difficulties Questionnaire; and cognitive impairment, which was measured by the mental processing composite scale of the Kaufman Assessment Battery for Children (an IQ-equivalent measure of cognitive ability in 2 dimensions: sequential and simultaneous processing scores). RESULTS: LDA treatment was administered to 125 of 584 (21%) mothers and was not significantly associated with mortality, cerebral lesions, cerebral palsy, or global cognitive impairment of the children at 5 years of age. The proportion of low simultaneous processing scores (<70) was lower in the group with LDA (7% vs 19% without LDA; P = .04). This association was not significant after adjustment for propensity score, prognostic factors, and social class (adjusted odds ratio [aOR]: 0.59 [95% confidence interval (CI): 0.17-2.06]). LDA treatment was associated with a reduction, at the limit of significance, in total behavioral difficulties (aOR: 0.44 [95% CI: 0.19-1.02]) and hyperactivity (aOR: 0.43 [95% CI: 0.17-1.05]). CONCLUSIONS: LDA was not associated with adverse neonatal or long-term outcomes. Moreover, the results suggest that LDA may be associated with a reduction in neurobehavioral difficulties. More research is needed to assess the effects of aspirin alone or combined with other neuroprotective agents. PMID- 20026501 TI - Immediate treatment versus sonographic surveillance for mild hip dysplasia in newborns. AB - OBJECTIVE: We conducted a blinded, randomized, controlled trial to examine whether mildly dysplastic but stable or instable hips would benefit from early treatment, as compared with watchful waiting. PATIENTS AND METHODS: A total of 128 newborns with mild hip dysplasia (sonographic inclination angle [alpha angle] of 43 degrees -49 degrees ) and stable or instable but not dislocatable hips were randomly assigned to receive either 6 weeks of abduction treatment (immediate treatment group) or follow-up alone (active-sonographic-surveillance group). The main outcome measurement was the acetabular inclination angle, measured by radiograph, at 1 year of age. RESULTS: Both groups included 64 newborns, and there was no loss to follow-up. With the exception of a small but statistically significant excess of girls in the active-sonographic-surveillance group, there were no statistically significant differences in baseline characteristics between the 2 groups. The mean inclination angle at 12 months was 24.2 degrees for both groups (difference: 0.1 [95% confidence interval (CI): -0.8 to 0.9]), and all children had improved and were without treatment. The mean alpha angle was 59.7 degrees in the treatment group and 57.1 degrees in the active-surveillance group for a difference of 2.6 degrees evaluated after 1.5 and 3 months (95% CI: 1.8 to 3.4; P < .001). At 1.5 months of age, the hips had improved in all treated children but not in 5 children under active surveillance (P = .06). Among the sonographic-surveillance group, 47% received treatment after the initial surveillance period of 1.5 months. CONCLUSIONS: Active-sonographic-surveillance halved the number of children requiring treatment, did not increase the duration of treatment, and yielded similar results at 1-year follow-up. Given a reported prevalence of 1.3% for mildly dysplastic but stable hips, a strategy of active surveillance would reduce the overall treatment rate by 0.6%. Our results may have important implications for families as well as for health care costs. PMID- 20026502 TI - Childhood polyarteritis nodosa in autoimmune lymphoproliferative syndrome. AB - Autoimmune lymphoproliferative syndrome (ALPS) is an uncommon disorder of Fas mediated apoptosis that results in impaired lymphocyte death and, therefore, disturbed immune homeostasis. Besides presentation with lymphadenopathy and splenomegaly, patients with ALPS have a high incidence of autoimmune phenomena. To our knowledge, this is the first description of polyarteritis nodosa that includes numerous arterial aneurysms in a child with ALPS. Active vasculitis resolved after allogeneic hematopoietic stem cell transplantation. This report of polyarteritis nodosa associated with human ALPS supports previous findings in Fas deficient mouse models that frequently develop vasculitic manifestations and suggests that apoptotic defects of lymphocytes may play a role in the pathophysiology of systemic vasculitis. Thus, patients with ALPS might be more susceptible to autoimmune vessel inflammation. This case furthermore emphasizes that even rare autoimmune manifestations should be considered and investigated in patients with immunodeficiencies, because that might help in planning treatment strategies for these patients. PMID- 20026504 TI - Isolation and characterization of microsatellite loci for Neothraupis fasciata, (Emberizidae, Passeriformes) with widely cross amplification in neotropical passerines. AB - We described the development and characterization of 16 microsatellite loci for the white-banded tanager (Neothraupis fasciata), a Neotropical passerine from Cerrado (Brazilian savanna). Based on 24 N. fasciata from a single population, we detected moderate to high number of alleles per locus (2-18 alelles) with expected heterozygosities ranging from 0.12 to 0.92. The high combined paternity exclusion probability (0.999) and low combined genetic identity (1.72 x 10(-10)) show that the battery of loci is suitable for parentage and population analyses. Some of these markers also showed amplification in 8 species from 4 Neotropical passerine families, indicating the potential of these markers to address similar questions in these species. PMID- 20026503 TI - Evaluation of biomarkers of exposure in adult cigarette smokers using Marlboro snus. AB - INTRODUCTION: It has been reported that adult smokers (AS) may be considering smokeless tobacco products as an alternative to smoking. The objective of this study was to evaluate the change in exposure in AS using Marlboro snus (MSNUS) (a tobacco pouch product in test market in June 2007). METHODS: AS were randomized into the following groups--CS: subjects (n = 30) continue smoking their own brand; DU: subjects (n = 60) reduced their daily cigarette consumption by >or=50% and were allowed to use MSNUS; SN: subjects (n = 15) stopped smoking their cigarettes but were allowed to use MSNUS; NT: subjects (n = 15) were not allowed to use any tobacco products for the entire duration of the 8-day study. Biomarkers of smoke exposure (BOE) measured at baseline and postbaseline were 24 hr urinary excretion of metabolites of N-nitrosamines, nicotine (urine and plasma), aromatic amines, benzene, and polycyclic aromatic hydrocarbon; urine mutagenicity; and carboxyhemoglobin at various timepoints. RESULTS: Statistically significant (p < .05) reductions in all the urinary BOE were observed in the DU group compared with the CS group. After correcting for the residual effect, a proportionate reduction (approximately 50%) in most of the biomarkers was observed. Even larger reductions, similar to the NT group, were observed in the SN group. DISCUSSION: The proportionate reduction in exposure when reducing the number of cigarettes by 50% and using MSNUS, under the consumption patterns observed, suggest that the AS did not appear to alter their smoking behavior. The added exposure from MSNUS usage in this group was minimal. The AS sustained substantial reductions in exposure when using MSNUS exclusively. PMID- 20026505 TI - A spanner in the herbal works. PMID- 20026508 TI - Bush insider recounts how ideology trumped science on US AIDS policy. PMID- 20026509 TI - WHO dispatches H1N1 vaccines to priority group of 35 poorer nations. PMID- 20026510 TI - New HIV drug patent pool "offers hope to millions". PMID- 20026511 TI - Swine flu has been "considerably less lethal than feared," says Donaldson. PMID- 20026513 TI - Physiological and cognitive mediators for the association between self-reported depressed mood and impaired choice stepping reaction time in older people. AB - BACKGROUND: The aim of the study was to use path analysis to test a theoretical model proposing that the relationship between self-reported depressed mood and choice stepping reaction time (CSRT) is mediated by psychoactive medication use, physiological performance, and cognitive ability. METHODS: A total of 280 retirement village residents, aged 62-95 years, undertook tests of CSRT, which required them to step onto one of four panels that were illuminated in a random order. Depressed mood was assessed using the 30-item Geriatric Depression Scale (GDS). The participants were also tested on physiological and cognitive performance, including quadriceps strength, balance, complex attention (Trail Making Test [TMT] B), simple reaction time, reported level of exercise, and use of psychoactive medications. RESULTS: A total of 51 participants (18%) showed mild to severe depression. Those with higher GDS scores had significantly increased CSRT and worse performance on all physiological and cognitive parameters. CSRT was also significantly associated with all other measures. The final path analysis model revealed an association between self-reported depression and CSRT that was mediated by two paths, one through quadriceps strength and the other through TMT B with both mediating variables then influencing CSRT via simple reaction time and balance. CONCLUSIONS: The findings suggest that self-reported depressed mood is related to slowed performance on a CSRT task and that this relationship is explained by underlying physiological and cognitive impairments. PMID- 20026514 TI - Arterio-venous shunts or low oxygen utilization? AB - An idea of arteriovenous shunts (AVS) was proposed for explanation of dynamic regulation of oxygenation and venous hyperoxia. A formula enabling calculation of AVS and real CO2 production has recently been derived by comparing data of arterial and venous blood gases. Regarding venous hyperoxia, there is a need to differentiate capillary to tissue transport defect (low oxygen utilisation-LOU) from AVS, which may exist simultaneously. The AVS may be associated with normal or relatively high oxygen utilization from the capillary vessels and increased CO2 production. AVS is proposed to carry protective and 'stealing' properties including renal, cardiac, and pulmonary hemodynamic. Calculations of the AVS may be important for dynamic assessment of vascular and metabolic status and in emergency medicine. PMID- 20026515 TI - Effects of combined exposure to dichlorvos and monocrotophos on blood and brain biochemical variables in rats. AB - Dichlorvos (DDVP) and monocrotophos (MC) are systemic insecticides and known to produce cholinergic and non-cholinergic effects. Individual toxic effects of these chemicals are known but their combined effects have not been studied. We studied the effect of concomitant exposure to DDVP and MC on selected biochemical variables suggestive of liver damage, changes in whole brain biogenic amines levels, acetylcholinesterase (AchE) and monoamine oxidase (MAO) activities in rats. Female rats were exposed to DDVP (2.5 mg/kg subcutaneously) and MC (1.8 mg/kg oral) either individually or in combination for 4 weeks. We observed significant decrease in more pronounced depletion in norepinephrine (NE) and dopamine (DA) levels during co-exposure to DDVP and MC. Brain AChE activity increased and activity of MAO showed significant depletion on co-exposure to DDVP and MC. Brain glutathione (GSH) and oxidized glutathione (GSSG) ratio decreased significantly during exposure to DDVP or MC while co-exposure to these toxicants led to a more pronounced depletion of GSH: GSSG ratio. Serum aspartate amino transferase (AST) and alkaline phosphatase (ALP) activities increased significantly on exposure to MC suggesting liver injury, while DDVP alone had no effect on these variables. There were no effects of DDVP and MC exposure on haematological biochemical variables except for depletion in serum glucose level after MC exposure which was more pronounced DDVP + MC during co-exposure. It can be concluded that only moderate synergistic effects occur between MC and DDVP during co-exposure. A more detailed study with variable doses, prolonged exposure and alterations in different brain regions is recommended. PMID- 20026516 TI - Ranitidine as an alcohol dehydrogenase inhibitor in acute methanol toxicity in rats. AB - Methanol poisoning is a hazardous intoxication characterized by visual impairment and formic acidemia. The therapy for methanol poisoning is alcohol dehydrogenase (ADH) inhibitors to prevent formate accumulation. Ranitidine has been considered to be an inhibitor of both gastric alcohol and hepatic aldehyde dehydrogenase enzymes. This study aimed at testing ranitidine as an antidote for methanol acute toxicity and comparing it with ethanol and 4-methyl pyrazole (4-MP). This study was conducted on 48 Sprague-Dawley rats, divided into 6 groups, with 8 rats in each group (one negative control group [C1], two positive control groups [C2, C3] and three test groups [1, 2 and 3]). C2, C3 and all test groups were exposed to nitrous oxide by inhalation, then, C3 group was given methanol (3 g/kg orally). The three test groups 1, 2 and 3 were given ethanol (0.5 g/kg orally), 4-MP (15 mg/kg intraperitoneally) and ranitidine (30 mg/kg intraperitoneally), respectively, 4 hours after giving methanol. Rats were sacrificed and heparinized, cardiac blood samples were collected for blood pH and bicarbonate. Non-heparinized blood samples were collected for formate levels by high performance liquid chromatography. Eye balls were enucleated for histological examination of the retina. Ranitidine corrected metabolic acidosis (p = .025), decreased formate levels (p = .014) and improved the histological findings in the retina induced by acute methanol toxicity. PMID- 20026517 TI - Pulse wave amplitude is associated with brachial artery diameter: implications for gender differences in microvascular function. AB - The ratio of pulse wave amplitude (PWA) during reactive hyperemia compared to baseline as measured by peripheral arterial tonometry (PAT) is a non-invasive measure of microvascular endothelial function referred to as the pulse wave amplitude reactive hyperemia index (PWA-RHI). Whether upstream conduit vessel structure may affect downstream resistance vessel PWA has not been clearly examined. We tested the hypothesis that digital PWA is influenced by brachial artery diameter (BAD) and that this association would influence comparison of PWA RHI between genders. Measures of vascular structure and microvascular function were carried out in 115 patients varying in cardiovascular risk profiles (average age 57 years, male n = 79, CAD n = 43). PWA was assessed using plethysmography at baseline and following 5 minutes of brachial artery occlusion. BAD was assessed using high-resolution ultrasonography. Results : There was a negative association between BAD and PWA-RHI ( r = -0.34, p < 0.05). Women had greater PWA-RHI and smaller BAD compared with men (p < 0.05). When co-varying for BAD, there were no longer gender differences in PWA-RHI. Moreover, when a sub-group of men and women without CAD (n = 40), matched for BAD, were examined, there were no gender differences in PWA-RHI. In conclusion, PWA-RHI obtained from PAT is associated with BAD. Studies examining gender differences in microvascular endothelial function with PAT may need to correct for BAD as a potential confounder. PMID- 20026520 TI - Metabolic syndrome in Argentinean patients with systemic lupus erythematosus. PMID- 20026519 TI - Neurocognitive deficits and neuroimaging abnormalities are prevalent in children with lupus: clinical and research experiences at a US pediatric institution. AB - Neurocognitive impairments and neuroimaging abnormalities are frequently observed in adults with systemic lupus erythematosus. There is a paucity of similar data in childhood-onset disease. We hypothesized that neurocognitive and neuroimaging abnormalities would be prevalent in children undergoing neuropsychological evaluations. We reviewed patient neurocognitive evaluations performed at a large United States pediatric institution during the period 2001 to 2008. Records were retrieved from 24 children referred to neuropsychology due to clinical indications. Data from 15 children enrolled in a prospective structure-function association study were also analyzed. Subjects were predominantly African American and Hispanic adolescent girls of average intelligence. aPL positivity and aspirin use was prevalent. Neurocognitive impairment was designated in 70.8% of retrospective, and 46.7% of prospective cohort patients. Deficits were seen at times of wellness, without previous neuropsychiatric lupus, and early in disease courses. Scores >1.5 standard deviations below published age-matched norms were common in tests of executive functioning, visual memory and visual-spatial planning. Features of depression were seen in 33.3% of the children in the retrospective cohort (clinical referrals). Cerebral and cerebellar volume loss was observed in a majority of blinded prospective cohort research magnetic resonance images (73.3% and 67.7% respectively). White matter hyperintensities were observed in retrospective and prospective cohort magnetic resonance images (36.6% and 46.7% respectively). Larger prospective studies that elucidate structure-function associations in children with systemic lupus erythematosus are planned. PMID- 20026521 TI - Pericarditis with cardiac tamponade in systemic lupus erythematosus. Development immediately following successful control of lupus flare. PMID- 20026522 TI - Hemophagocytic syndrome as one of the main primary manifestations in acute systemic lupus erythematosus--case report and literature review. AB - Hemophagocytic syndrome is an unusual but fatal disorder characterized by pancytopenia and activation of macrophages. We describe one case of acute systemic lupus erythematosus with an unusual presentation of hemophagocytic syndrome not related to infection. The patient presented with pancytopenia related to increasing hemophagocytic activity of histiocytes in the bone marrow. Concomitant class IV World Health Organization lupus nephritis, serositis, high titer of antinuclear factor and positive test for anti-DNA antibody fitted the diagnostic criteria of systemic lupus erythematosus. She also presented with alveolar hemorrhage and lupus myocarditis. She underwent immunosuppressive therapy with recovery from the hemophagocytic syndrome. Therefore, diagnosis of acute lupus hemophagocytic syndrome was made. The clinical presentation, laboratory diagnosis, and management of the patient are discussed and the literature was reviewed and presented, with emphasis on a possible distinct lupus subset, which includes a more aggressive systemic disease with heart involvement. PMID- 20026523 TI - Systemic lupus erythematosus and strongyloidiasis: a multifaceted connection. AB - We describe a case of systemic lupus erythematosus complicated by strongyloidiasis. The parasitic infection appeared with diarrhoea, weight loss and peripheral eosinophilia in association with recurrence of polyarthritis, probably due to a flare of systemic lupus erythematosus. The literature about the coexistence of systemic lupus erythematosus and strongyloidiasis has been reviewed. PMID- 20026524 TI - Identification of autoantigens specific for systemic lupus erythematosus with central nervous system involvement. AB - Using proteomic analysis, we identified candidate autoantigens specific for central nervous system (CNS) involvement in systemic lupus erythematosus (SLE). Proteins, extracted from cultured human neuroblastoma cells, were separated both by SDS-PAGE (1-DE) and two-dimensional electrophoresis (2-DE), and transferred to membranes. Western blot analysis was performed using serum samples from 30 SLE patients with CNS involvement (CNS-Lupus) and from 30 SLE patients without CNS involvement (non-CNS-SLE). The detected autoantigens were identified using MALDI TOF/TOF MS. On the 1-DE Western blot, we detected 32 antigenic bands in the serum samples from the CNS-Lupus patients. Among them, four bands were detected significantly more frequently in the CNS-Lupus patients than in the non-CNS-SLE patients. Three bands were detected in four or more of the CNS-Lupus patients but in only one or none of the non-CNS-SLE patients. We thus selected these seven bands for the next investigations. Next, we detected protein spots corresponding to the selected seven bands by 2-DE Western blot and identified four proteins. They are peroxiredoxin-4, ubiquitin carboxyl-terminal hydrolase isozyme L1, splicing factor arginine/serine-rich 3, and histone H2A type 1. These four candidate autoantigens for the anti-neuronal cell antibodies would be a useful marker for CNS-Lupus. PMID- 20026525 TI - Neglecting the importance of the decision making and care regimes of personal support workers: a critique of standardization of care planning through the RAI/MDS. AB - PURPOSE: The Resident Assessment Instrument-Minimum Data Set (RAI/MDS) is an interdisciplinary standardized process that informs care plan development in nursing homes. This standardized process has failed to consistently result in individualized care planning, which may suggest problems with content and planning integrity. We examined the decision making and care practices of personal support workers (PSWs) in relation to the RAI/MDS standardized process. DESIGN AND METHODS: This qualitative study utilized focus groups and semi structured interviews with PSWs (n = 26) and supervisors (n = 9) in two nursing homes in central Canada. RESULTS: PSWs evidenced unique occupational contributions to assessment via proximal familiarity and biographical information as well as to individualizing care by empathetically linking their own bodily experiences and forging bonds of fictive kinship with residents. These contributions were neither captured by RAI/MDS categories nor relayed to the interdisciplinary team. Causal factors for PSW exclusion included computerized records, low status, and poor interprofessional collaboration. Intraprofessional collaboration by PSWs aimed to compensate for exclusion and to individualize care. IMPLICATIONS: Exclusive institutional reliance on the RAI/MDS undermines quality care because it fails to capture residents' preferences and excludes input by PSWs. Recommendations include incorporating PSW knowledge in care planning and documentation and examining PSWs' nascent occupational identity and their role as interprofessional brokers in long-term care. PMID- 20026526 TI - The exceptional ethics of the investigator-subject relationship. AB - This article concerns the validity of six canonical rules that institutional review boards use to constrain the behavior of investigators. These rules require investigators to design their studies in a scientifically valid way, not pay their subjects to take risks, minimize risks to their subjects, secure for their subjects access to effective interventions post-trial, not pay their subjects too much and allow their subjects to withdraw from the study unconditionally. Enforcement of these rules is problematic because there are other relationships that seem to be like the investigator-subject relationship in all ethically relevant respects, such as the employer-employee and volunteer organizer volunteer relationships, to which we would not dream of applying these same rules. Applying these rules in one context but not the others is a violation of ethical consistency I label "exceptionalism." We should conclude that it is time to reexamine the validity of the six rules. PMID- 20026527 TI - The proper role of evidence in complementary/alternative medicine. AB - In this article we explore the role evidence ought to play in complementary and alternative medicine (CAM). First, we consider the claim that evidence in the form of randomized controlled trials (RCTs) cannot be obtained for CAMs. Second, we consider various claims to the effect that there are ways of obtaining evidence that do not make use of RCTs. We argue that there is no good reason why CAM should be exempted from the general requirement that treatments undergo evaluation by RCT. Third, we consider two implications for health care policy. First, many activities in conventional medicine have never been rigorously evaluated and are widely in use nonetheless. We argue that this fails to provide a reason for exempting CAM from a demand for evidence. Second, CAM use may be compared to a choice of lifestyle, and this has a significant impact on which requirements of evidence can reasonably be imposed. PMID- 20026528 TI - La frontera: responsibly managing borders and boundaries in clinical ethics. AB - The papers in the 2010 "Clinical Ethics" number of the Journal of Medicine and Philosophy explore issues along La Frontera, the borders and boundaries of clinical ethics. The first three papers in this "Clinical Ethics" number of the Journal explore borders and boundaries drawn within clinical ethics, concerning the moral standing of complementary and alternative medicine, palliative sedation, and induced abortion and feticide. The fourth and fifth papers explore the borders and boundaries between research ethics and clinical ethics. PMID- 20026529 TI - Endovascular management of Stanford type A (ascending) aortic dissection. PMID- 20026530 TI - Identifying paraplegia risk associated with thoracic endografting. AB - Endografting, like open surgical repair of the thoracic aorta, can be complicated by paraplegia. We reviewed our thoracic endografting experience regarding the incidence and treatment of spinal cord neurologic events. Between February 2000 and July 2008, 346 patients underwent endoluminal grafting of the descending thoracic aorta. Indications for intervention included atherosclerotic aneurysms (45.9%), acute and chronic dissections (31.5%), miscellaneous lesions (13.6%), and penetrating aortic ulcers (8.9%). Ten women and 4 men (4.0%), with a mean age of 71.3 years, developed either paraparesis (1.7%) or paraplegia (2.3%). Nine (64.3%) of these patients had an aneurysm, 4 (28.6%) had acute or chronic type B aortic dissection, and 1 (7.1%) had a penetrating aortic ulcer; 3 (21.4%) of them had previously undergone open abdominal aortic aneurysm repair, and 13 (92.9%) required coverage of more than 20 cm of the aorta. Cerebrospinal fluid drainage was instituted in 7/8 paraplegic patients. Eight (57.1%) of the 14 patients recovered fully, 2 (14.3%) experienced partial recovery, and 4 (28.6%) had significant neurological deficits. Paraplegia following thoracic endografting appears to be associated with female sex, long-segment coverage of the thoracic aorta, and aneurysmal disease. PMID- 20026531 TI - Cardiac bioassist: results of the French multicenter cardiomyoplasty study. AB - The French multicenter experience (6 centers) of dynamic cardiomyoplasty was analyzed for long-term survival and functional outcome, the most important endpoints in congestive heart failure therapy. Cardiomyoplasty was performed in 212 patients with symptoms of chronic heart failure despite maximal pharmacological therapy. The etiology was ischemic (48%), idiopathic (45%) or other (7%). Cardiomyoplasty was performed using the latissimus dorsi muscle which was electrostimulated after surgery. During follow-up, 88% of patients improved clinically. Hospital death occurred in 29 (14%) patients and was related to the severity of preoperative heart failure symptoms. Late mortality occurred in 99 patients due to heart failure (44%), sudden death (37%), or noncardiac causes (18%). Combined dynamic cardiomyoplasty and implantation of a cardiac rhythm management system was safely achieved in 22 patients, and 26 underwent heart transplantation for recurrent heart failure. Long-term functional improvements were observed in most patients, and the best outcome was achieved in those with isolated right ventricular failure. Dynamic cardiomyoplasty can be considered as a destination therapy or a mid- to long-term biological bridge to heart transplantation. PMID- 20026532 TI - Stem cell experiments and initial clinical trial of cellular cardiomyoplasty. AB - Growing myocardial cells from human stem cells and stem cell transplantation to repair injured myocardium are new frontiers in cardiovascular research. The 1st stage of this study was conducted to determine whether transplantation of autologous bone marrow stem cells into infarcted myocardium of sheep could differentiate into beating cardiomyocytes. The 2nd stage was to demonstrate transdifferentiation of human bone marrow mesenchymal stem cells to precursor cardiomyocytes in vitro, using a novel conditioning medium. In the 3rd stage, a clinical trial of stem cell implantation in patients with severe myocardial dysfunction involved injection of peripheral blood-derived endothelial precursor cells in 11 patients and autologous bone marrow mononuclear cells in 29. A marginal improvement in myocardial function was noted at 3 months (mean increase in ejection fraction, 6% +/- 1%), although it plateaued at 6 months. The trial proved to be safe because there was no procedure-related mortality. There is growing optimism that stem cell therapy may delay heart transplantation. PMID- 20026533 TI - Figure-of-eight vs. interrupted sternal wire closure of median sternotomy. AB - Sternal dehiscence is a rare but devastating complication following median sternotomy for cardiac surgery. The optimal technique for sternal closure is unclear. We conducted this prospective randomized trial to compare the incidence of sternal dehiscence after figure-of-8 and simple interrupted suturing in patients undergoing coronary artery bypass grafting. Between January 2007 and June 2008, 98 patients had figure-of-8 suturing and 97 had interrupted sutures. The mean age of the patients was 60.9 +/- 7.6 years. The overall sternal dehiscence rate was 8%; 7 cases in the in figure-of-8 group and 9 in the interrupted group. Thirteen patients had no wound infection and healed with conservative treatment. Only 3 patients had sternal dehiscence with infection: 2 with simple interrupted closure and 1 with figure-of-8 sternal closure. There was no significant difference in rates of sternal dehiscence between the 2 groups. It was concluded that figure-of-8 sternal suturing is equally effective as simple interrupted suturing in preventing sternal dehiscence. PMID- 20026534 TI - Impact of valves in a biomechanical heart model assisting failing hearts. AB - Experimental valveless muscular blood pumps (biomechanical hearts) in goats can pump more than 1 L.min(-1), but due to a high pendulum volume, no significant flow contribution to the circulation is gained. Thus valved and valveless biomechanical hearts were compared for efficacy. Heart failure was induced in 5 adult Bore goats by repeated intracoronary embolization. A valved and balloon equipped pumping chamber was integrated into the descending aorta, simulating standard biomechanical circulatory support. The valveless biomechanical heart supported a failing heart with a baseline cardiac output of 2,670 +/- 710 mL.min( 1) by contributing additional flow of 113 +/- 37 mL.min(-1). The biomechanical heart model incorporating an outlet valve offered an additional 304 +/- 126 mL.min(-1), and the use of 2 valves significantly enhanced pulmonary blood flow by 1,235 +/- 526 mL.min(-1). The use of 2 valves in biomechanical hearts seems to be essential to achieve adequate circulatory support. Double-valved biomechanical hearts driven by an appropriate skeletal muscle ventricle may contribute to the therapy of heart failure. PMID- 20026535 TI - Blunt traumatic bronchial rupture in patients younger than 18 years. AB - The aim of this study was to evaluate our results of management of bronchial rupture after blunt chest trauma in young patients. Between January 2000 and December 2007, 34 patients aged 6-18-years old were treated for bronchial rupture; 25 presented early and 9 presented late. The 21 (62%) boys and 13 (38%) girls had a mean age of 11.58 +/- 2.51 years. Radiography, computed tomography, and rigid bronchoscopy were performed in stable acute cases and late-presenting cases, with rapid bronchoscopy followed by thoracotomy in urgent cases. In the patients who presented early, repair was undertaken through a right thoracotomy in 21 (84%) and via a left thoracotomy in 4 (16%). Direct suturing was carried out in all except 3 patients who needed lobectomy. Reimplantation of the main bronchus after debridement of the edges was possible in 4 (44%) of the chronic cases, and pulmonary resection was required in 5 (56%). Four (11.8%) patients died due to perioperative cardiac arrest and cerebral hypoxia with failure of resuscitation. Meticulous collaboration between the surgeon and anesthetist is important to ensure survival. PMID- 20026536 TI - Prolonged QT interval and coronary artery bypass mortality due to heart failure. AB - QT-interval prolongation has been shown to predict mortality in coronary artery disease and heart failure. To assess the prognostic value of QT interval for death due to low cardiac output after coronary artery bypass grafting, the QT interval was measured in 3 consecutive beats on the preoperative electrocardiogram (leads II and V(4)) in 30 patients who died perioperatively due to heart failure and a control group of 168 randomly matched hospital survivors during the same 3-year period. Mean corrected QT interval was significantly longer in the patients who died compared to the control group (480.7 +/- 96.2 vs. 425.4 +/- 21 ms). Among the variables evaluated, QT prolongation was the only independent predictor of perioperative death. In patients admitted for coronary artery bypass grafting, QT interval measurement is a simple clinical tool that may identify patients with a greater probability of a troublesome operative course. PMID- 20026537 TI - Etiopathology and management challenges of blunt chest trauma in Nigeria. AB - Blunt chest trauma had not been roundly studied in Nigeria. This study was conducted to determine the etiopathological and management challenges of chest trauma in a developing country. In a prospective multicenter hospital-based study of 10-years duration in the city of Lagos, the data of 896 patients were recorded. The male-to-female ratio was 8 : 1, and the mean age was 27.1 years. The majority of patients (76.9%) were aged 20-39 years. Road traffic accidents caused 98.1% of the injuries. Lung parenchymal injuries occurred in 66% of patients, and cardiac trauma in 0.1%. Isolated chest trauma was found in 85% of patients, and 134 had associated injuries. The incidence of blunt chest trauma could be reduced if the number of road traffic accidents in Nigeria is reduced. PMID- 20026538 TI - Risk factors for mediastinitis and endocarditis after cardiac surgery. AB - A prospective open-cohort study was performed in 838 adults undergoing coronary revascularization or valve surgery to define the risk factors for development of surgical site infections. Patients diagnosed with mediastinitis or endocarditis during follow-up were compared with patients with no such infection. After 1 year of follow-up, 22 (2.6%) patients had developed mediastinitis or endocarditis. No preoperative or intraoperative variables were identified as risk factors. By multivariate analysis of postoperative variables, respiratory insufficiency, microorganisms in blood cultures, and intensive care unit stay were independent risk factors for the development of these complications. The type of antibiotic prophylaxis had no influence on the incidence of organ or space infections after cardiac surgery. PMID- 20026539 TI - Incidence and impact of cardiac cachexia in valvular surgery. AB - Cardiac cachexia is a wasting feature of advanced heart failure, which is due to several etiologies and associated with a poor prognosis. This study assessed the incidence and impact of cardiac cachexia retrospectively in 353 patients who underwent valve surgery from June 2005 to June 2006. Using 80% of ideal body weight as a cut-off point, 46 (13%) of these patients were considered to have cardiac cachexia. Patients with cachexia were predominantly male with more underlying diseases and a lower body mass index than those without cachexia. The New York Heart Association functional class was significantly worse in patients with cachexia (class III/IV: 54.4% vs. 21.2%) and they had greater incidences of active infective endocarditis (21.7% vs. 5.2%) and tricuspid regurgitation (41.3% vs. 21.8%) compared to those with normal body weight. The cachexia group had significantly longer postoperative hospitalization and more complications (37% vs. 21.5%); perioperative mortality tended to be higher (6.5% vs. 2.3%) although not statistically significant. Cardiac cachexia remains an important problem in patients undergoing valve surgery, which indicates end-staged disease, and contributes to poor perioperative outcomes. Special care and attention are needed in this particular group of patients. PMID- 20026540 TI - Integrated positron-emission tomography for nodal staging in lung cancer. AB - As lymph node metastasis is the most important factor determining the surgical outcome of lung cancer, we evaluated the accuracy and clinical usefulness of functional imaging with integrated positron-emission tomography and computed tomography in nodal staging of non-small-cell lung cancer. Between October 2003 and August 2004, 43 newly diagnosed patients underwent curative surgical resection or mediastinoscopic nodal biopsy after chest computed tomography as well as whole-body functional imaging for clinical staging. Based on histopathologic findings in the lymph nodes, we compared the diagnostic usefulness of computed tomography and functional imaging. Twenty-two patients were N0 stage, 6 were N1, 11 were N2, and 4 were N3. Although functional imaging seemed to be superior for evaluation of N2 nodes, there were no significant differences in the sensitivity, specificity, positive-predictive value, negative predictive value, or accuracy. The situation was similar for N1 nodes, and there were no significant differences except for the false-negative rate. High false positive and negative-rates for N2 lymph nodes suggest that functional imaging should not replace mediastinoscopy. PMID- 20026541 TI - Long-term results up to 19 years of mitral balloon valvuloplasty. AB - Clinical and echocardiographic data of 547 consecutive patients (mean age, 31.5 years) undergoing mitral balloon valvuloplasty with follow-up of 1.5-19 years, were analyzed. Immediately after valvuloplasty, mitral valve area increased significantly from 0.92 +/- 0.17 to 1.95 +/- 0.29 cm(2). Restenosis occurred in 169 (31%) patients; it was less common (20%) in those with a mitral echocardiographic score 8 and preexisting atrial fibrillation as predictors of combined events. Valvuloplasty provides excellent results in selected patients with mitral stenosis. The long-term outcome can be predicted from the baseline characteristics of the mitral valve. PMID- 20026542 TI - Malignancy of congenital cystic adenomatoid malformation of lung in aged. AB - Congenital cystic adenomatoid malformation is rare in adults, and often asymptomatic. We describe a case of malignant degeneration of a congenital cystic adenomatoid malformation in a 77-year-old man. Congenital cystic adenomatoid malformation must be considered a premalignant lesion, even in older patients. Rigorous preoperative staging must be undertaken. Anatomic resection with extensive lymphadenectomy remains the treatment of choice. PMID- 20026543 TI - Aneurysm of all 3 sinuses of valsalva causing coronary insufficiency. AB - A 35-year-old woman presented with chest pain and easy fatigability. Transthoracic echocardiography demonstrated an unruptured aneurysm involving all 3 sinuses of Valsalva. Successful surgical repair was undertaken, without the need for aortic valve replacement or coronary artery bypass. PMID- 20026544 TI - Type A aortic dissection, right-sided aortic arch, and thoracic aortic aneurysm. AB - A previously healthy 78-year-old woman presented with severe chest pain. Computed tomography revealed a right-sided aortic arch with an aberrant left subclavian artery originating from Kommerell's diverticulum, a thoracic aortic aneurysm, and Stanford type A acute aortic dissection. Successful emergency repair of the ascending aorta, total arch, and descending aorta were performed through a median sternotomy only. PMID- 20026545 TI - Repairing of infarcted myocardium using a novel cell patch therapy. AB - A 54-year-old man underwent left anterior descending artery bypass grafting with cell patch therapy for myocardial infarction. We applied a novel approach to replicating the infarcted myocardium by anchoring polyglycolic acid felt seeded with autologous peripheral blood mononuclear cells in front of the infarcted area. The cell patch remained fixed to the myocardium for 1 month, and the infarcted myocardium was reperfused. PMID- 20026546 TI - Late presentation of aortic root abscess in endocarditis with coronary ischemia. AB - Formation of a large aortic root abscess is an infrequent complication of aortic valve endocarditis in adults. Extrinsic compression of the coronary arteries by this abscess is still rarer. Here, we report a case of a 22-year-old male with aortic root abscess, who presented 2 months after the completion of treatment of endocarditis with exertional angina. Coronary angiogram revealed compression of proximal left anterior descending and left circumflex arteries by the abscess. The patient was successfully treated with pericardial patch exclusion of the abscess cavity and coronary artery bypass graft. The presentation of aortic root abscess with myocardial ischemia as a late complication of treated endocarditis has not been reported earlier. PMID- 20026547 TI - Beating-heart valve surgery: A systematic review. AB - Beating-heart continuous coronary perfusion (BHCCP) has been promoted as an alternative to the technique of cardioplegic arrest in valve surgery. Its potential advantage is the elimination of cardioplegia and the corollary risk of ischemic reperfusion injury. The use of CCP has been recommended especially when performing more complex operations, such as mitral valve repair, and particularly as surgeons become more familiar with beating-heart coronary surgery. We conducted a systematic review to assess the strength of the evidence supporting the efficacy of BHCCP compared to cardioplegia in valve surgery. Thirty nine reports were identified. Of these, only two were randomized control trials. Overall the studies were generally of poor quality and had a low evidence level. In those studies, mortality and major morbidity from BHCCP were within acceptable levels, nevertheless, there was no advantage over cardioplegic arrest. On the other hand there is weak evidence that it may reduce functional and biochemical markers of myocardial injury. In conclusion, BHCCP is an operative strategy in valve surgery with some potential benefits. There is, however a need for a high quality, prospective, randomized control trial to establish the exact role for BHCCP in complex valve surgery. PMID- 20026548 TI - Giant cystic lymphangioma in the thoracic wall in a newborn. PMID- 20026549 TI - Heart and great vessels after mediastinal irradiation. PMID- 20026550 TI - Contrast-enhanced computed tomography of adult scimitar syndrome (variant form). PMID- 20026551 TI - Chronic pulmonary hypertension due to in-situ pulmonary thrombus. PMID- 20026552 TI - Predictors of the quality of care for asthma in general practice: an observational study. AB - BACKGROUND: Asthma is a common and important health condition in the UK, predominantly managed in primary care. Little is known about how characteristics of practices and patients are associated with achievement of quality indicators (QIs) for asthma. OBJECTIVE: To measure the recorded quality of primary care for asthma and to assess whether quality of care differed by patient and practice characteristics. METHODS: Medical records were examined for 253 randomly selected asthma patients from 18 general practices in England. Quality of care was assessed against seven predetermined QIs. Logistic regression models were used to test variations in quality of care by age, gender, patient postcode deprivation rank, practice size and time point. RESULTS: There was substantial variation in achievement of individual QIs (range 39-97%). Participants whose postcodes were in the most deprived areas were more likely to be asked about difficulties sleeping [odds ratios (ORs) 1.7, 95% confidence interval (CI) 1.2-2.5] or whether asthma interfered with daily activities (OR 1.8, CI 1.2-2.7) than those from middle or least deprived postcode areas. QIs were more likely to be achieved in 2005 than 2003 (ORs 4.4, 2.4, 3.0). There were no significant differences by other characteristics. CONCLUSIONS: Great variations exist in the quality of primary care for asthma and considerable scope for improvement. Asthma care improved over time. The preliminary findings that quality of asthma care varied with deprivation support the idea that primary care may be targeting care to those in most need. However, variations were small and only significant for two QIs. PMID- 20026553 TI - Acute non-traumatic hip pathology in children: incidence and presentation in family practice. AB - BACKGROUND: The differential diagnosis of children with acute non-traumatic hip pathology varies from quite harmless conditions such as transient synovitis of the hip to more severe problems like Perthes' disease, slipped capital femoral epiphysis (SCFE) and life-threatening conditions such as septic arthritis of the hip. OBJECTIVE: To provide population-based data on symptom presentation and incidence rates of non-traumatic acute hip pathology in family practice. METHODS: We analysed data from a large national survey of family practice (104 practices), which was carried out by the Netherlands Institute for Health Services Research (NIVEL) in 2001. We included all children aged 0-14 years. Incidence rates were calculated by dividing the total number of cases (numerator) by the average study population at risk (denominator). RESULTS: Our study population consisted of 73 954 children aged 0-14 years, yielding 68 202 person-years. These children presented with 101 episodes of acute non-traumatic hip pathology. The presenting feature in 81.5% of the children was pain, in 8.6% limping and 9.9% presented with both symptoms. Only 27% of the participating family practitioners (FPs) reported whether the child had a fever. The incidence rate for all acute non traumatic hip pathology was 148.1 per 100 000 person-years, and for transient synovitis, this was 76.2 per 100 000 person-years. CONCLUSION: In family practice, most children with acute non-traumatic hip pathology present with pain as the initial symptom. FPs need to be more aware that fever is the main distinguishing factor between a harmless condition and a life-threatening condition. Transient synovitis is the diagnosis with the highest incidence rate. PMID- 20026554 TI - Effect of nutritional counselling and nutritional plus exercise counselling in overweight adults: a randomized trial in multidisciplinary primary care practice. AB - OBJECTIVE: To compare the effects of nutritional counselling with nutritional plus exercise counselling on body weight and waist circumference in overweight adults in a multidisciplinary primary care setting. METHODS: One hundred and thirty-four overweight adults (body mass index 28-35) were randomly assigned to individual counselling sessions by a dietician (D) or counselling sessions by a dietician plus physiotherapist (D + E) during 6 months with one follow-up session at 12 months. Outcomes were assessed at baseline, 6 and 12 months. Difference in changes of outcome measures between groups were analysed using generalized estimating equations. RESULTS: Weight reduced from baseline to 6 months in D [ 2.2 (-3.1 to -1.4) kg] and D + E [-3.0 (-4.0 to -2.0) kg] and was sustained at 12 months [-2.0 (-3.1 to -1.4) kg and -3.1 (-4.5 to -1.6) kg, respectively]. The reduction in weight did not significantly differ between D and D + E (P = 0.48). In both groups, waist circumference decreased from baseline to 6 months [-2.1 ( 3.3 to -0.8) cm for D; -3.7 (-5.1 to -2.3) cm for D + E] and was sustained at 12 months [-2.1 (-3.5 to -0.7) cm and -4.2 (-6.0 to -2.5) cm, respectively]. Participants in D + E tended to decrease their waist circumference more than those in D (P = 0.14). DISCUSSION: Nutritional counselling by a dietician resulted in modest reductions in weight and waist circumference in overweight adults, which were sustained up to 12 months. Adding exercise counselling by a physiotherapist did not significantly enhance the effect on body weight. Exercise counselling may, however, further improve waist circumference. PMID- 20026555 TI - Copy number variations in East-Asian population and their evolutionary and functional implications. AB - Recent discovery of the copy number variation (CNV) in normal individuals has widened our understanding of genomic variation. However, most of the reported CNVs have been identified in Caucasians, which may not be directly applicable to people of different ethnicities. To profile CNV in East-Asian population, we screened CNVs in 3578 healthy, unrelated Korean individuals, using the Affymetrix Genome-Wide Human SNP array 5.0. We identified 144,207 CNVs using a pooled data set of 100 randomly chosen Korean females as a reference. The average number of CNVs per genome was 40.3, which is higher than that of CNVs previously reported using lower resolution platforms. The median size of CNVs was 18.9 kb (range 0.2 5406 kb). Copy number losses were 4.7 times more frequent than copy number gains. CNV regions (CNVRs) were defined by merging overlapping CNVs identified in two or more samples. In total, 4003 CNVRs were defined encompassing 241.9 Mb accounting for approximately 8% of the human genome. A total of 2077 CNVRs (51.9%) were potentially novel. Known CNVRs were larger and more frequent than novel CNVRs. Sixteen percent of the CNVRs were observed in > or =1% of study subjects and 24% overlapped with the OMIM genes. A total of 476 (11.9%) CNVRs were associated with segmental duplications. CNVS/CNVRs identified in this study will be valuable resources for studying human genome diversity and its association with disease. PMID- 20026556 TI - A Drosophila mutant of LETM1, a candidate gene for seizures in Wolf-Hirschhorn syndrome. AB - Human Wolf-Hirschhorn syndrome (WHS) is a multigenic disorder resulting from a hemizygous deletion on chromosome 4. LETM1 is the best candidate gene for seizures, the strongest haploinsufficiency phenotype of WHS patients. Here, we identify the Drosophila gene CG4589 as the ortholog of LETM1 and name the gene DmLETM1. Using RNA interference approaches in both Drosophila melanogaster cultured cells and the adult fly, we have assayed the effects of down-regulating the LETM1 gene on mitochondrial function. We also show that DmLETM1 complements growth and mitochondrial K(+)/H(+) exchange (KHE) activity in yeast deficient for LETM1. Genetic studies allowing the conditional inactivation of LETM1 function in specific tissues demonstrate that the depletion of DmLETM1 results in roughening of the adult eye, mitochondrial swelling and developmental lethality in third instar larvae, possibly the result of deregulated mitophagy. Neuronal specific down-regulation of DmLETM1 results in impairment of locomotor behavior in the fly and reduced synaptic neurotransmitter release. Taken together our results demonstrate the function of DmLETM1 as a mitochondrial osmoregulator through its KHE activity and uncover a pathophysiological WHS phenotype in the model organism D. melanogaster. PMID- 20026557 TI - Successful treatment of lipoid pneumonia associated with bowel obstruction by Ascaris lumbricoides. AB - Partial bowel obstruction is a serious complication of ascariasis infestation generally treated with mineral oil. This prospective study aimed to evaluate the efficacy of multiple bronchoalveolar lavages (BAL) as a therapeutic strategy for reducing lung inflammation of lipoid pneumonia associated with ascariasis. The study included five children (mean age 25 months) with partial small-bowel obstruction by Ascaris lumbricoides, who underwent diagnostic bronchoalveolar lavage for assessment of refractory pneumonia. Routine biochemical, microbiological and cytological analysis were carried out in the BAL. Protein, lactate dehydrogenase and tumor necrosis factor-alpha (TNF-alpha) cytokine levels were determined in the serum before and after treatment. At admission, children consistently had respiratory symptoms, altered hematological function, increased immunoglobulin E serum level and peripheral blood eosinophilia. Chest tomography showed consolidation with air bronchogram (4/4), ground-glass infiltration (3/4) and decreased attenuation in the consolidation areas (2/4). Presence of marked pleocytosis with Sudan positive foamy alveolar macrophages, high protein and lactate dehydrogenase levels in the BAL indicated presence of mixed alveolitis. One child with extensive consolidation and air bronchogram in both lungs died before treatment. Multiple bronchoalveolar lavages efficiently removed alveolar oil deposits, restored BAL cellularity, improved clinical symptoms, radiological parameters and further reduced inflammatory reaction evidenced by marked decrease of the inflammatory cytokine, TNF-alpha. This study presents a therapeutic strategy for management of lung complications caused by mineral oil administration to treat intestinal bowel obstruction associated with ascariasis. PMID- 20026559 TI - Cortical thickness reduction in individuals at ultra-high-risk for psychosis. AB - Although schizophrenia is characterized by gray matter (GM) abnormalities, particularly in the prefrontal and temporal cortices, it is unclear whether cerebral cortical GM is abnormal in individuals at ultra-high-risk (UHR) for psychosis. We addressed this issue by studying cortical thickness in this group with magnetic resonance imaging (MRI). We measured cortical thickness of 29 individuals with no family history of psychosis at UHR, 31 patients with schizophrenia, and 29 healthy matched control subjects using automated surface based analysis of structural MRI data. Hemispheric mean and regional cortical thickness were significantly different according to the stage of the disease. Significant cortical differences across these 3 groups were found in the distributed area of cerebral cortices. UHR group showed significant cortical thinning in the prefrontal cortex, anterior cingulate cortex, inferior parietal cortex, parahippocampal cortex, and superior temporal gyrus compared with healthy control subjects. Significant cortical thinning in schizophrenia group relative to UHR group was found in all the regions described above in addition with posterior cingulate cortex, insular cortex, and precentral cortex. These changes were more pronounced in the schizophrenia group compared with the control subjects. These findings suggest that UHR is associated with cortical thinning in regions that correspond to the structural abnormalities found in schizophrenia. These structural abnormalities might reflect functional decline at the prodromal stage of schizophrenia, and there may be progressive thinning of GM cortex over time. PMID- 20026558 TI - Cognition in mouse models of schizophrenia susceptibility genes. AB - Cognitive deficits are core features of psychiatric disorders and contribute substantially to functional outcome. It is still unclear, however, how cognitive deficits are related to underlying genetic liability and overt clinical symptoms. Fortunately, animal models of susceptibility genes can illuminate how the products of disease-associated genetic variants affect brain function and ultimately alter behavior. Using as a reference findings from the Cognitive Neuroscience Treatment Research to Improve Cognition in Schizophrenia program and the SchizophreniaGene database, we review cognitive data from mutant models of rare and common genetic variants associated with schizophrenia. PMID- 20026560 TI - Transition to the adult nephrologist does not induce acute renal transplant rejection. AB - BACKGROUND: In spite of the overall increased renal graft survival, long-term allograft survival has remained least successful in adolescent recipients. A major change in their care is the transition from the paediatric to the adult nephrology unit. METHODS: To analyse the effect of transition on the acute rejection frequency and graft survival, we performed a historical cohort study in all patients transplanted at the paediatric unit between 1980 and 2004. Data were obtained by reviewing medical charts in two of the four Dutch pediatric renal transplantation centers from time of transplantation until 3 years after transition. For analysis, we used a Cox proportional hazards model. RESULTS: The cohort consisted of 162 patients: 133 native Dutch and 29 immigrant patients. Transition occurred at a mean age of 18 years (range 14-22). At transition, 72% had a functioning allograft. Acute rejections occurred in 92/162 patients before (median follow-up 4.8 years, range 0.2-12.8) and in 15/116 patients after transition (median follow-up 3.0 years, range 1.6-3.0). Most rejections (62%) occurred within the first year after transplantation. The relative risk of acute rejections after transition in comparison to before transition was 0.10 [95% confidence interval (95% CI) 0.04-0.28] in Dutch patients and 0.69 (95% CI 0.33 1.40) in immigrant patients. In the 3 years before transition, 28/154 patients (18%) experienced graft failure compared to 19/116 patients (16%) in the 3 years after transition. CONCLUSIONS: The risk for acute rejection decreases after transition to the adult unit. There is less risk reduction in immigrant patients. Nephrologists should pay special attention to these patients. PMID- 20026561 TI - Tropical rheumatology in a UK District General Hospital: a case report of leprosy presenting as acute vasculitis. PMID- 20026562 TI - Hand bone mineral density is associated with both total hip and lumbar spine bone mineral density in post-menopausal women with RA. AB - OBJECTIVE: RA is associated with localized bone loss in the hands, as well as generalized osteoporosis. We evaluated the relationship between hand digital X ray radiogrammetry BMD (DXR-BMD) and total hip and lumbar spine BMD. METHODS: We conducted a cross-sectional study of 138 post-menopausal women with RA. The DXR BMD was calculated based on digitized hand radiographs. Measurements of the total hip and lumbar spine BMD were performed by a DXA-BMD (BMDa) scan. Patient and physician questionnaires and laboratory samples supplied information on relevant covariates. Separate multivariate linear regression models were constructed to determine the cross-sectional relationship between hand DXR-BMD (independent variable) and total hip or lumbar spine BMD (dependent variables). RESULTS: The cohort comprised women with a median age of 61 years and RA disease duration of 13 years. Seventy-six per cent were either RF and/or anti-cyclic citrullinated peptide (anti-CCP) positive and most had moderate disease activity [median disease activity score-28 joint count (DAS28) 3.7]. Hand DXR-BMD was significantly associated with total hip BMD (beta = 0.61; P < 0.0001) and lumbar spine BMD (beta = 0.62; P < 0.0008) in adjusted models. CONCLUSIONS: This study suggests that hand DXR-BMD is associated with both the total hip and lumbar spine BMD among post-menopausal women with RA. The relationship between bone loss in the hands and generalized osteoporosis should be further explored in longitudinal studies of patients with RA. PMID- 20026563 TI - Towards understanding the phenotypes of myocardial involvement in the presence of self-limiting and sustained systemic inflammation: a magnetic resonance imaging study. AB - OBJECTIVE: To investigate the patterns of myocardial involvement in the presence of self-limiting and sustained systemic inflammation, using MRI. METHODS: Ninety four subjects, with a clinical diagnosis of myocarditis (n = 36), RA (n = 24) and apparently healthy subjects (n = 34, control group), underwent standardized cardiac MRI protocol for the assessment of global and regional morphology and systolic function using balanced steady-state free precession sequences, T2 weighted images and late gadolinium enhancement (LGE) studies. RESULTS: The three groups were well matched for age, gender and cardiovascular risk factors. The RA group showed markedly increased end-diastolic volumes and reduced ejection fraction (P < 0.05). Antero/inferolateral wall thickness was greater in the myocarditis group and reduced in RA, associated with reduced radial and longitudinal thickening (P < 0.01), and markedly raised T2-oedema ratio and global LGE scores (P < 0.05). CONCLUSIONS: Our results may signify the phenotypic features of myocardial plasticity and deformation in response to self-limiting and sustained inflammatory injury. PMID- 20026564 TI - Serum levels of IL-33 and soluble ST2 and their association with disease activity in systemic lupus erythematosus. AB - OBJECTIVE: IL-33 has recently been found to be the specific ligand of ST2, an IL 1 receptor family member that is selectively expressed in Th2 cells and mediates Th2 response. This study aims to measure the serum levels of soluble ST2 (sST2) and IL-33 in patients with SLE and to examine their association with disease activity. METHODS: Seventy SLE patients were evaluated for disease activity, determined by SLEDAI, levels of anti-dsDNA antibody, C3 and C4. Fifty-seven patients were evaluated longitudinally on a second occasion. IL-33 and sST2 were measured by sandwich ELISA in the 127 SLE serum samples and compared with 28 age- and sex-matched healthy controls. RESULTS: Serum sST2 level was significantly higher in active SLE patients [0.51 (0.18) ng/ml] compared with inactive patients [0.42 (0.08) ng/ml] (P = 0.006) and normal controls [0.36 (0.13) ng/ml] (P < 0.001). sST2 level correlated significantly with SLEDAI, anti-dsDNA antibody and prednisolone dosage, and negatively with C3. Linear regression analysis showed that serum sST2 level was an independent predictive factor for modified SLEDAI, excluding anti-dsDNA and complement score after controlling for age, sex, glomerular filtration rate and prednisolone dosage (regression coefficient: 8.5; 95% CI 2.6, 14.3) (P = 0.005). Serum sST2 level was sensitive to change in disease activity longitudinally, with an effect size of 0.29. Elevated serum IL 33 was comparable in frequency (4.3 vs 7.1%; P = 0.62) and levels (P = 0.53) between SLE patients and controls. CONCLUSIONS: Elevated serum sST2 level in SLE patients was found to correlate with disease activity and was sensitive to change, suggesting a potential role as a surrogate marker of disease activity. PMID- 20026566 TI - Editorial: Integrating research and practice: the role of the Journal of Pediatric Psychology. PMID- 20026565 TI - The relationship among child weight status, psychosocial functioning, and pediatric health care expenditures in a medicaid population. AB - OBJECTIVES: To examine the association between weight status and health service use, while considering the influence of psychosocial functioning and demographic variables. METHODS: Two hundred child-parent dyads were recruited from pediatric primary care clinics and completed measures of height, weight, and questionnaires assessing psychosocial functioning. Claims and expenditure data over a 12-month retrospective period were extracted from the Medicaid claims database. RESULTS: Children who were obese incurred greater health service use and expenditures than children who were of a healthy weight, even after controlling for psychosocial functioning and other demographic variables. Children who were overweight (but not obese) did not have differing levels of claims or expenditures than their healthy weight peers. CONCLUSIONS: Understanding the impact of pediatric obesity on long-term expenditures is critical. These results provide some indication of the financial savings that might be achieved if obese children were supported to achieve a healthier weight status. PMID- 20026567 TI - Brief report: Barriers to treatment adherence in pediatric inflammatory bowel disease. AB - OBJECTIVE: To examine perceived barriers to medication adherence in inflammatory bowel disease (IBD) treatment and their relationship with adherence using a combined forced choice and semi-structured interview assessment approach. METHODS: Sixteen adolescents with IBD and their parents participated in an open ended interview regarding adherence barriers and completed quantitative measures of adherence, barriers to treatment, and disease severity. RESULTS: The most commonly identified barriers to adherence were forgetting, interference with other activities, difficulty swallowing pills, and not being at home. Number of reported barriers was positively correlated with objective nonadherence for 6 MP/azathioprine. Nonadherence frequency was 42% for 6-MP/azathoprine and 50% for 5-ASA medications. CONCLUSIONS: Using a combined assessment approach, patients and parents reported several barriers to treatment adherence that are appropriate for clinical intervention. This is critical given the significant medication nonadherence observed in this sample and the relationship between total number of barriers and disease management problems. PMID- 20026568 TI - Using structural equation modeling to understand child and parent perceptions of asthma quality of life. AB - OBJECTIVE: Using structural equation modeling, test a conceptual model of associations between constructs predicting parent and child asthma quality of life. METHODS: Children with a confirmed asthma diagnosis and their parents completed measures of health status and independently reported on psychological functioning, family functioning, and quality of life. RESULTS: Measurement and structural models for predicting parent and child quality of life provided a good fit of data to the conceptual model. Parent and child independent reports of quality of life are dependent upon family functioning and child psychological functioning. Long-term asthma symptom control is the only health status variable that impacts quality of life. CONCLUSIONS: With minor modifications, both parent and child data fit the conceptual model. Child psychological functioning and long term asthma control jointly contribute to quality of life outcomes. Findings suggest that both acute and long-term asthma health status outcomes have different determinants. PMID- 20026569 TI - A camp-based intervention targeting independence among individuals with spina bifida. AB - OBJECTIVE: To design and evaluate a camp-based intervention, the goal of which was to increase independence among children, adolescents, and adults with spina bifida. METHODS: An intervention targeting independence was embedded within a typical week long camp experience. The intervention consisted of the following: collaborative (i.e., parent and camper) goal identification, group sessions consisting of psycho-education and cognitive tools, and goal monitoring by camp counselors. Camper and parent report of demographic variables, goal attainment, spina bifida knowledge, and independence were gathered. Interventionist report of adherence to the treatment manual was also collected. RESULTS: Campers made significant gains in individual goals, management of spina bifida responsibilities, and independence with general spina bifida tasks, with medium effect sizes observed in goal attainment. CONCLUSIONS: Results indicated that significant progress was made on individually oriented goals from pre- to post camp. Design issues are discussed. PMID- 20026570 TI - Relationships between somatic growth and cognitive functioning in young children with sickle cell disease. AB - OBJECTIVE: Children with sickle cell disease (SCD) exhibit poor somatic growth due to nutritional and metabolic effects, but potential relationships between growth and other areas of development are unclear. We examined whether growth is related to cognition and whether growth might be one marker of neurocognitive risk. METHODS: Sixty-four children with SCD and eighty-one demographically similar controls, ages 4 to 8 years, completed cognitive and anthropometric measures. RESULTS: Height-for-age partially accounted for cognitive decrements related to SCD on all cognitive measures. Higher body-mass-index was a significant predictor of higher visual-motor and academic achievement scores in children with SCD, but not in controls. CONCLUSIONS: In some children with SCD, especially those with HbSS and Hb Sbeta(0), low height-for-age may help to explain neurocognitive risk. Higher body-mass-index may be related to better cognitive outcomes in children with SCD. Nutrition deficits in SCD could explain the association between somatic growth and cognitive deficits. PMID- 20026571 TI - A community-based exercise and education scheme for stroke survivors: a randomized controlled trial and economic evaluation. AB - OBJECTIVE: The evaluation of a community-based exercise and education scheme for stroke survivors. DESIGN: A single blind parallel group randomized controlled trial. SETTING: Leisure and community centres in the south-west of England. SUBJECTS: Stroke survivors (median (IQR) time post stroke 10.3 (5.4-17.1) months). 243 participants were randomized to standard care (124) or the intervention (119). INTERVENTION: Exercise and education schemes held twice weekly for eight weeks, facilitated by volunteers and qualified exercise instructors (supported by a physiotherapist), each with nine participants plus carers or family members. METHOD: Participants were assessed by a blinded independent assessor at two weeks before the start of the scheme, nine weeks and six months. One-year follow-up was by postal assessment. MAIN MEASURES: PRIMARY OUTCOMES: Subjective Index of Physical and Social Outcome (SIPSO); Frenchay Activities Index; Rivermead Mobility Index. NHS, social care and personal costs. Secondary outcomes included WHOQoL-Bref. ANALYSIS: Intention-to-treat basis, using non-parametric analysis to investigate change from baseline. Economic costs were compared in a cost-consequences analysis. RESULTS: There were significant between-group changes in SIPSO physical at nine weeks (median (95% confidence interval (CI)), 1 (0, 2): P = 0.022) and at one year (0 (-1, 2): P = 0.024). (WHOQol-Bref psychological (6.2 (-0.1, 9.1): P = 0.011) at six months. Mean cost per patient was higher in the intervention group. The difference, excluding inpatient care, was pound296 (95% CI: - pound321 to pound913). CONCLUSION: The community scheme for stroke survivors was a low-cost intervention successful in improving physical integration, maintained at one year, when compared with standard care. PMID- 20026572 TI - Exercise on a treadmill or walking outdoors? A randomized controlled trial comparing effectiveness of two walking exercise programmes late after stroke. AB - OBJECTIVE: To evaluate spatial and temporal gait characteristics and endurance late after stroke in people who had received two different walking exercises. A secondary aim was to compare the outcomes in relation to length of time exercising and number of exercise occasions between the two. DESIGN: A randomized controlled trial. SETTING: A private rehabilitation centre. SUBJECTS: Thirty-nine people with stroke entered the study, and five dropped out. INTERVENTIONS: Treadmill training versus walking outdoors. MAIN MEASURES: Six-Minute Walk Test, a 10-metre walk test and pulse rates at rest and in activity. RESULTS: There were significant differences in favour of the treadmill group in Six-Minute Walk Test distance (P = 0.04), Six-Minute Walk Test speed (P = 0.03), 10-m walking speed (P = 0.03), bilateral stride length (right leg; P = 0.009, left leg; P = 0.003) and step width (P = 0.01), indicating more symmetrical use of the legs in the treadmill group (1.02-1.10 m versus 0.97-0.92 m). There were no significant differences between groups in cadence (P = 0.78). All participants complied 100% with their respective programmes. Exercise frequency did not differ between the groups but significantly less time was spent exercising on the treadmill compared with walking exercise outdoors (107 versus 316 minutes, P = 0.002). There were no differences in use of assistive aids between the groups on arrival at the clinic or at departure. CONCLUSION: The results indicate that treadmill walking improves spatial and temporal gait characteristics more effectively than walking outdoors. PMID- 20026573 TI - Goal attainment scaling: a direct comparison of alternative rating methods. AB - PURPOSE: To compare two alternative rating systems for goal attainment scaling (GAS) intended to improve capture of goal underachievement. SETTING: A tertiary inpatient neurorehabilitation unit. DESIGN: Secondary comparative analysis of prospective cohort data. PARTICIPANTS: A total of 243 patients were included, with 977 goals between them; M:F ratio 146:97, mean age 44.8 (SD 14.2) years. DIAGNOSIS: 200 (82%) acquired brain injury, 21 (9%) spinal cord injury, 22 (9%) other neurological conditions. METHODS: Goal attainment scaling-rated achievement of 1-6 patient-selected goals was collected using the standard 5-point scale (-2 to +2) and formula to derive aggregated T-scores, as recommended by its originators. Two alternative 6-point rating systems were applied retrospectively. Version 1 set all baseline scores at '-2' and added '-3' to denote 'worsening'. Version 2 added a '-0.5' score to denote 'partial achievement' for goals starting at '-1', without change to range or baseline scores. RESULTS: Baseline scores were identical for standard rating and version 2 (median 34.9, interquartile range (IQR) 31.9-35.5), but lower for version 1 (median 21.0, IQR 19.8-22.6), which was reflected in the change scores. While median achieved T-scores were 50.0 for all three methods, version 2 marginally overestimated goal attainment (Wilcoxon z -6.8, P < 0.001), while version 1 underestimated it (Wilcoxon z -7.2, P < 0.001), in comparison with standard goal rating. CONCLUSION: Different goal rating methods may have significant impact on goal attainment scaling results. Version 2 provided the closest match to standard rating, supporting documentation of partial goal achievement without affecting score range. For analysis and reporting, '0.5' scores should be converted to '-1', to maintain parity with standard rating systems. PMID- 20026574 TI - Meeting stroke survivors' perceived needs: a qualitative study of a community based exercise and education scheme. AB - OBJECTIVE: A qualitative study using a phenomenological approach, to explore stroke survivors' needs and their perceptions of whether a community stroke scheme met these needs. METHODS: Semi-structured in-depth interviews of 12 stroke survivors, purposively selected from participants attending a new community stroke scheme. Interpretative phenomenological analysis of interviews by two researchers independently. RESULTS: Participants attending the community stroke scheme sought to reconstruct their lives in the aftermath of their stroke. To enable this they needed internal resources of confidence and sense of purpose to 'create their social self', and external resources of 'responsive services' and an 'informal support network', to provide direction and encouragement. Participants felt the community stroke scheme met some of these needs through exercise, goal setting and peer group interaction, which included social support and knowledge acquisition. CONCLUSION: Stroke survivors need a variety of internal and external resources so that they can rebuild their lives positively post stroke. A stroke-specific community scheme, based on exercise, life-centred goal setting, peer support and knowledge acquisition, is an external resource that can help with meeting some of the stroke survivor's needs. PMID- 20026575 TI - Evaluation of an adjustment group for people with multiple sclerosis: a pilot randomized controlled trial. AB - OBJECTIVE: The aim was to evaluate a group treatment for people with multiple sclerosis and low mood. DESIGN: Randomized controlled trial. SETTING: Community. PARTICIPANTS: Patients with multiple sclerosis and low mood, scoring >7 on the Hospital Anxiety and Depression Scales or >2 on the General Health Questionnaire 12. INTERVENTIONS: Participants either attended an adjustment group for six, 2 hour group treatment sessions or were on a waiting list to attend the group. OUTCOMES: Hospital Anxiety and Depression Scale, General Health Questionnaire 12, Multiple Sclerosis Self Efficacy Scale, Multiple Sclerosis Impact Scale and Short Form 36 administered 3 and 6 months after random allocation. RESULTS: Of the 219 patients identified, 100 (46%) reported depressive symptoms and 126 (58%) anxiety symptoms. Forty participants were recruited, aged 25-68 (mean 47.7 SD 9.7) and eight were men. Patients allocated to the group intervention reported fewer depressive symptoms than those in the control group (U 109.5, P<0.05) but there were no significant differences in anxiety symptoms, self-efficacy or quality of life. CONCLUSION: Depressive symptoms were reduced following group intervention, which suggests this may be an effective psychological treatment and warrants further evaluation. PMID- 20026576 TI - Therapy to improve gestural expression in aphasia: a controlled clinical trial. AB - OBJECTIVES: To assess whether production of communicative gestures can be ameliorated by therapy, to explore factors that might influence individual variability of therapeutic success and to find out whether therapy effects generalize to unpractised gestures. SETTING: Hospital department specializing in rehabilitation of patients with neuropsychological disorders. SUBJECTS: Twenty three consecutive severely aphasic patients received therapy; ten were tested repeatedly without interpolated therapy. DESIGN: Twenty-four communicative gestures were divided into three sets of eight gestures. In the therapy group one set was practised during each of three consecutive periods and after that period changes in proficiency were assessed for all three sets. Comparisons were made between changes of practised and unpractised gestures during each therapy period within the therapy group, and between changes of unpractised gestures in the therapy group, and the changes brought forward by repeated testing in the control group. INTERVENTIONS: Therapy was centred on familiarizing patients with the communicative functions of gestures and on improving the spatial precision of their gestures. MAIN MEASURE: Scoring of gestures considered their accuracy and the necessity of assistance. RESULTS: Therapy led to substantial improvement of practised gestures. There was also improvement of unpractised gestures which was significantly smaller than that of practised gestures but greater than the improvement induced by repeated testing in the control group. CONCLUSIONS: In view of the limited generalization, gesture therapy should concentrate on gestures that are relevant for communication in daily living of the individual patients. PMID- 20026577 TI - The use of measurement tools in clinical practice: an observational study of neurorehabilitation. AB - OBJECTIVE: To explore the way in which standardized measurement tools are used in day-to-day clinical practice. DESIGN: Non-participant observation and semi structured interviews. SETTING: Hospital-based adult neurorehabilition unit. PARTICIPANTS: The multidisciplinary team delivering rehabilitation in the unit. RESULTS: The multidisciplinary team used the measurement tools internally to establish the presence and severity of patients' impairments and activity limitations; predict recovery and discharge destination; inform treatment planning and monitor progress. They were used externally to demonstrate service effectiveness; communicate with patients, families and external agencies; enable other providers to plan continuing care and resource needs, and inform audit and service development. CONCLUSION: When used in everyday clinical practice by a multidisciplinary neurorehabilitation team, standardized measures were used to support and inform, rather than replace, clinical decision-making. PMID- 20026578 TI - An integrated programme after pulmonary rehabilitation in patients with chronic obstructive pulmonary disease: effect on emotional and functional dimensions of quality of life. AB - OBJECTIVE: To assess whether a maintenance integrated health care programme is effective in improving functional and emotional dimensions of quality of life in patients with chronic obstructive pulmonary disease (COPD) after a first pulmonary rehabilitation. DESIGN: Prospective controlled trial. SETTING: Three rehabilitation centres and three patient self-help associations within a health care network in France. SUBJECTS: Forty patients with moderate to severe COPD. INTERVENTIONS: After a first four-week inpatient pulmonary rehabilitation programme, patients took part in a maintenance integrated health care programme or usual care for 12 months. MAIN MEASURES: The primary outcomes were the change in functional and emotional dimensions of quality of life measured by the St George's Respiratory Questionnaire (SGRQ), the brief World Health Organization Quality of Life questionnaire (Brief-WHOQOL) and six specific questions using a 10-cm visual analogue scale. Secondary outcomes were change in exercise tolerance measured by six-minute walking test and cycle exercise. RESULTS: At one year, the maintenance intervention (n = 11) produced improvements in functional and emotional dimensions scores of quality of life and exercise tolerance. Patients in the usual aftercare group (n = 16) exhibited maintenance of functional dimension scores of quality of life, but a clinically relevant decline in emotional scores of quality of life and in six-minute walking distance one year after the pulmonary rehabilitation. CONCLUSION: Patient self-help association seems to be an innovative and efficient organizational structure to support patients with COPD after pulmonary rehabilitation in real-life settings. A distinction between emotional and functional dimensions of quality of life may improve the design and evaluation of integrated health care programmes in patients with COPD. PMID- 20026579 TI - Comparing 3 dietary pattern methods--cluster analysis, factor analysis, and index analysis--With colorectal cancer risk: The NIH-AARP Diet and Health Study. AB - The authors compared dietary pattern methods-cluster analysis, factor analysis, and index analysis-with colorectal cancer risk in the National Institutes of Health (NIH)-AARP Diet and Health Study (n = 492,306). Data from a 124-item food frequency questionnaire (1995-1996) were used to identify 4 clusters for men (3 clusters for women), 3 factors, and 4 indexes. Comparisons were made with adjusted relative risks and 95% confidence intervals, distributions of individuals in clusters by quintile of factor and index scores, and health behavior characteristics. During 5 years of follow-up through 2000, 3,110 colorectal cancer cases were ascertained. In men, the vegetables and fruits cluster, the fruits and vegetables factor, the fat-reduced/diet foods factor, and all indexes were associated with reduced risk; the meat and potatoes factor was associated with increased risk. In women, reduced risk was found with the Healthy Eating Index-2005 and increased risk with the meat and potatoes factor. For men, beneficial health characteristics were seen with all fruit/vegetable patterns, diet foods patterns, and indexes, while poorer health characteristics were found with meat patterns. For women, findings were similar except that poorer health characteristics were seen with diet foods patterns. Similarities were found across methods, suggesting basic qualities of healthy diets. Nonetheless, findings vary because each method answers a different question. PMID- 20026580 TI - Age at menarche and risk of type 2 diabetes: results from 2 large prospective cohort studies. AB - The authors investigated the association between age at menarche and risk of type 2 diabetes mellitus (T2DM) among 101,415 women from the Nurses' Health Study (NHS) aged 34-59 years (1980-2006) and 100,547 women from Nurses' Health Study II (NHS II) aged 26-46 years (1991-2005). During 2,430,274 and 1,373,875 person years of follow-up, respectively, 7,963 and 2,739 incident cases of T2DM were documented. Young age at menarche was associated with increased risk of T2DM after adjustment for potential confounders, including body figure at age 10 years and body mass index (BMI; weight (kg)/height (m)(2)) at age 18 years. Relative risks of T2DM across age-at-menarche categories (< or =11, 12, 13, 14, and > or =15 years) were 1.18 (95% confidence interval (CI): 1.10, 1.27), 1.09 (95% CI: 1.02, 1.17), 1.00 (referent), 0.92 (95% CI: 0.83, 1.01), and 0.95 (95% CI: 0.84, 1.06), respectively, in the NHS (P for trend < 0.0001) and 1.40 (95% CI: 1.24, 1.57), 1.13 (95% CI: 1.00, 1.27), 1.00 (referent), 0.98 (95% CI: 0.82, 1.18), and 0.96 (95% CI: 0.78, 1.19), respectively, in NHS II (P for trend < 0.0001). Associations were substantially attenuated after additional control for updated time-varying BMI. These data suggest that early menarche is associated with increased risk of T2DM in adulthood. The association may be largely mediated through excessive adult adiposity. The association was stronger among younger women, supporting a role for sex hormones in younger onset of T2DM, in addition to BMI. PMID- 20026581 TI - The multi-domain protein Np95 connects DNA methylation and histone modification. AB - DNA methylation and histone modifications play a central role in the epigenetic regulation of gene expression and cell differentiation. Recently, Np95 (also known as UHRF1 or ICBP90) has been found to interact with Dnmt1 and to bind hemimethylated DNA, indicating together with genetic studies a central role in the maintenance of DNA methylation. Using in vitro binding assays we observed a weak preference of Np95 and its SRA (SET- and Ring-associated) domain for hemimethylated CpG sites. However, the binding kinetics of Np95 in living cells was not affected by the complete loss of genomic methylation. Investigating further links with heterochromatin, we could show that Np95 preferentially binds histone H3 N-terminal tails with trimethylated (H3K9me3) but not acetylated lysine 9 via a tandem Tudor domain. This domain contains three highly conserved aromatic amino acids that form an aromatic cage similar to the one binding H3K9me3 in the chromodomain of HP1ss. Mutations targeting the aromatic cage of the Np95 tandem Tudor domain (Y188A and Y191A) abolished specific H3 histone tail binding. These multiple interactions of the multi-domain protein Np95 with hemimethylated DNA and repressive histone marks as well as with DNA and histone methyltransferases integrate the two major epigenetic silencing pathways. PMID- 20026582 TI - DNA supercoiling and its role in DNA decatenation and unknotting. AB - Chromosomal and plasmid DNA molecules in bacterial cells are maintained under torsional tension and are therefore supercoiled. With the exception of extreme thermophiles, supercoiling has a negative sign, which means that the torsional tension diminishes the DNA helicity and facilitates strand separation. In consequence, negative supercoiling aids such processes as DNA replication or transcription that require global- or local-strand separation. In extreme thermophiles, DNA is positively supercoiled which protects it from thermal denaturation. While the role of DNA supercoiling connected to the control of DNA stability, is thoroughly researched and subject of many reviews, a less known role of DNA supercoiling emerges and consists of aiding DNA topoisomerases in DNA decatenation and unknotting. Although DNA catenanes are natural intermediates in the process of DNA replication of circular DNA molecules, it is necessary that they become very efficiently decatenated, as otherwise the segregation of freshly replicated DNA molecules would be blocked. DNA knots arise as by-products of topoisomerase-mediated intramolecular passages that are needed to facilitate general DNA metabolism, including DNA replication, transcription or recombination. The formed knots are, however, very harmful for cells if not removed efficiently. Here, we overview the role of DNA supercoiling in DNA unknotting and decatenation. PMID- 20026583 TI - Genome-wide localization analysis of a complete set of Tafs reveals a specific effect of the taf1 mutation on Taf2 occupancy and provides indirect evidence for different TFIID conformations at different promoters. AB - In Saccharomyces cerevisiae, TFIID and SAGA principally mediate transcription of constitutive housekeeping genes and stress-inducible genes, respectively, by delivering TBP to the core promoter. Both are multi-protein complexes composed of 15 and 20 subunits, respectively, five of which are common and which may constitute a core sub-module in each complex. Although genome-wide gene expression studies have been conducted extensively in several TFIID and/or SAGA mutants, there are only a limited number of studies investigating genome-wide localization of the components of these two complexes. Specifically, there are no previous reports on localization of a complete set of Tafs and the effects of taf mutations on localization. Here, we examine the localization profiles of a complete set of Tafs, Gcn5, Bur6/Ncb2, Sua7, Tfa2, Tfg1, Tfb3 and Rpb1, on chromosomes III, IV and V by chromatin immunoprecipitation (ChIP)-chip analysis in wild-type and taf1-T657K mutant strains. In addition, we conducted conventional and sequential ChIP analysis of several ribosomal protein genes (RPGs) and non-RPGs. Intriguingly, the results revealed a novel relationship between TFIIB and NC2, simultaneous co-localization of SAGA and TFIID on RPG promoters, specific effects of taf1 mutation on Taf2 occupancy, and an indirect evidence for the existence of different TFIID conformations. PMID- 20026584 TI - DNA stretching in the nucleosome facilitates alkylation by an intercalating antitumour agent. AB - DNA stretching in the nucleosome core can cause dramatic structural distortions, which may influence compaction and factor recognition in chromatin. We find that the base pair unstacking arising from stretching-induced extreme minor groove kinking near the nucleosome centre creates a hot spot for intercalation and alkylation by a novel anticancer compound. This may have far reaching implications for how chromatin structure can influence binding of intercalator species and indicates potential for the development of site selective DNA-binding agents that target unique conformational features of the nucleosome. PMID- 20026585 TI - Guanine sulphinate is a major stable product of photochemical oxidation of DNA 6 thioguanine by UVA irradiation. AB - The DNA of patients taking the immunosuppressant and anticancer drugs azathioprine or 6-mercaptopurine contains 6-thioguanine (6-TG). The skin of these patients is selectively sensitive to ultraviolet A radiation (UVA) and they suffer an extremely high incidence of sunlight-induced skin cancer with long-term treatment. DNA 6-TG interacts with UVA to generate reactive oxygen species, which oxidize 6-TG to guanine sulphonate (G(SO3)). We suggested that G(SO3) is formed via the reactive electrophilic intermediates, guanine sulphenate (G(SO)) and guanine sulphinate (G(SO2)). Here, G(SO2) is identified as a significant and stable UVA photoproduct of free 6-TG, its 2'-deoxyribonucleoside, and DNA 6-TG. Mild chemical oxidation converts 6-TG into G(SO2), which can be further oxidized to G(SO3)-a stable product that resists further reaction. In contrast, G(SO2) is converted back to 6-TG under mild conditions. This suggests that cellular antioxidant defences might counteract the UVA-mediated photooxidation of DNA 6-TG at this intermediate step and ameliorate its biological effects. In agreement with this possibility, the antioxidant ascorbate protected DNA 6-TG against UVA oxidation and prevented the formation of G(SO3). PMID- 20026586 TI - Fusion of short telomeres in human cells is characterized by extensive deletion and microhomology, and can result in complex rearrangements. AB - Telomere fusion is an important mutational event that has the potential to lead to large-scale genomic rearrangements of the types frequently observed in cancer. We have developed single-molecule approaches to detect, isolate and characterize the DNA sequence of telomere fusion events in human cells. Using these assays, we have detected complex fusion events that include fusion with interstitial loci adjacent to fragile sites, intra-molecular rearrangements, and fusion events involving the telomeres of both arms of the same chromosome consistent with ring chromosome formation. All fusion events were characterized by the deletion of at least one of the telomeres extending into the sub-telomeric DNA up to 5.6 kb; close to the limit of our assays. The deletion profile indicates that deletion may extend further into the chromosome. Short patches of DNA sequence homology with a G:C bias were observed at the fusion point in 60% of events. The distinct profile that accompanies telomere fusion may be a characteristic of the end joining processes involved in the fusion event. PMID- 20026587 TI - Generation of redesigned homing endonucleases comprising DNA-binding domains derived from two different scaffolds. AB - Homing endonucleases have become valuable tools for genome engineering. Their sequence recognition repertoires can be expanded by modifying their specificities or by creating chimeric proteins through domain swapping between two subdomains of different homing endonucleases. Here, we show that these two approaches can be combined to create engineered meganucleases with new specificities. We demonstrate the modularity of the chimeric DmoCre meganuclease previously described, by successfully assembling mutants with locally altered specificities affecting both I-DmoI and I-CreI subdomains in order to create active meganucleases with altered specificities. Moreover these new engineered DmoCre variants appear highly specific and present a low toxicity level, similar to I SceI, and can induce efficient homologous recombination events in mammalian cells. The DmoCre based meganucleases can therefore offer new possibilities for various genome engineering applications. PMID- 20026588 TI - Effects of abasic sites on structural, thermodynamic and kinetic properties of quadruplex structures. AB - Abasic sites represent the most frequent lesion in DNA. Since several events generating abasic sites concern guanines, this damage is particularly important in quadruplex forming G-rich sequences, many of which are believed to be involved in several biological roles. However, the effects of abasic sites in sequences forming quadruplexes have been poorly studied. Here, we investigated the effects of abasic site mimics on structural, thermodynamic and kinetic properties of parallel quadruplexes. Investigation concerned five oligodeoxynucleotides based on the sequence d(TGGGGGT), in which all guanines have been replaced, one at a time, by an abasic site mimic (dS). All sequences preserve their ability to form quadruplexes; however, both spectroscopic and kinetic experiments point to sequence-dependent different effects on the structural flexibility and stability. Sequences d(TSGGGGT) and d(TGGGGST) form quite stable quadruplexes; however, for the other sequences, the introduction of the dS in proximity of the 3'-end decreases the stability more considerably than the 5'-end. Noteworthy, sequence d(TGSGGGT) forms a quadruplex where dS does not hamper the stacking between the G tetrads adjacent to it. These results strongly argue for the central role of apurinic/apyrimidinic site damages and they encourage the production of further studies to better delineate the consequences of their presence in the biological relevant regions of the genome. PMID- 20026593 TI - RE: A further plea for adherence to the principles underlying science in general and the epidemiologic enterprise in particular. PMID- 20026589 TI - RNF4 and VHL regulate the proteasomal degradation of SUMO-conjugated Hypoxia Inducible Factor-2alpha. AB - Hypoxia-inducible factors (HIFs) are critical transcription factors that mediate cell survival during reduced oxygen conditions (hypoxia). At regular oxygen conditions (normoxia), HIF-1alpha and HIF-2alpha are continuously synthesized in cells and degraded via the ubiquitin-proteasome pathway. During hypoxia, these proteins are stabilized and translocate to the nucleus to activate transcription of target genes that enable cell survival at reduced oxygen levels. HIF proteins are tightly regulated via post-translational modifications including phosphorylation, acetylation, prolyl-hydroxylation and ubiquitination. Here we show for the first time that exogenous and endogenous HIF-2alpha are also regulated via the ubiquitin-like modifier small ubiquitin-like modifiers (SUMO). Using mutational analysis, we found that K394, which is situated in the sumoylation consensus site LKEE, is the major SUMO acceptor site in HIF-2alpha. Functionally, sumoylation reduced the transcriptional activity of HIF-2alpha. Similar to HIF-1alpha, HIF-2alpha is regulated by the SUMO protease SENP1. The proteasome inhibitor MG132 strongly stabilized SUMO-2-conjugated HIF-2alpha during hypoxia but did not affect the total level of HIF-2alpha. The ubiquitin E3 ligases von Hippel-Lindau and RNF4 control the levels of sumoylated HIF-2alpha, indicating that sumoylated HIF-2alpha is degraded via SUMO-targeted ubiquitin ligases. PMID- 20026594 TI - House value as an indicator of cumulative wealth is strongly related to morbidity and mortality risk in older people: a census-based cross-sectional and longitudinal study. AB - BACKGROUND: There has been relatively little research into health inequalities in older populations. This may be partly explained by the difficulty in identifying appropriate indicators of socio-economic status for older people. Ideally, indicators of socio-economic status to be used in studies of health inequalities in older populations should incorporate some measure of life-time socio-economic standing, and house value may fill this role. This study examined whether an indicator of accumulated wealth based on a combination of housing tenure and house value was a strong predictor of ill-health in older populations. METHODS: A total of 191 848 people aged > or =65 years and not living in communal establishments were identified from the 2001 Northern Ireland Census and followed for 5 years. Self-reported health and mortality risk by housing tenure/house value groupings were examined while controlling for a range of other demographic and socio-economic characteristics. RESULTS: Housing tenure/house value was highly correlated with other indicators of socio-economic status. Public-sector renters had worse self-reported health and higher mortality rates than owner occupiers but significant gradients were also found between those living in the highest- and lowest-valued owner-occupier properties. The relationship between housing tenure and value was unchanged by adjustment for indicators of social support and quality of the physical environment. Adjustment for limiting long term illness and self-reported health at baseline narrowed but did not eliminate the health gains associated with living in more expensive housing. CONCLUSIONS: House value of residence is an accessible and powerful indicator of accumulated wealth that is highly correlated with current health status and predictive of future mortality risk in older populations. PMID- 20026595 TI - Meta-analyses involving cross-over trials: methodological issues. PMID- 20026596 TI - 2009 influenza A(H1N1) monovalent vaccines for children. PMID- 20026597 TI - Immunogenicity of a monovalent 2009 influenza A(H1N1) vaccine in infants and children: a randomized trial. AB - CONTEXT: In the ongoing influenza pandemic, a safe and effective vaccine against 2009 influenza A(H1N1) is needed for infants and children. OBJECTIVE: To assess the immunogenicity and safety of a 2009 influenza A(H1N1) vaccine in children. DESIGN, SETTING, AND PARTICIPANTS: Randomized, observer-blind, age-stratified, parallel group study assessing 2 doses of an inactivated, split-virus 2009 influenza A(H1N1) vaccine in 370 healthy infants and children aged 6 months to less than 9 years living in Australia. INTERVENTION: Intramuscular injection of 15 microg or 30 microg of hemagglutinin antigen dose of monovalent, unadjuvanted 2009 influenza A(H1N1) vaccine in a 2-dose regimen, administered 21 days apart. MAIN OUTCOME MEASURES: Hemagglutination inhibition assay to estimate the proportion of participants with antibody titers of 1:40 or greater, seroconversion, or a significant antibody titer increase, and factor increase in geometric mean titer. Assessments of solicited adverse events during 7 days and unsolicited adverse events for 21 days after each vaccination. RESULTS: Following the first dose of vaccine, antibody titers of 1:40 or greater were observed in 161 of 174 infants and children in the 15-microg group (92.5%; 95% confidence interval [CI], 87.6%-95.6%) and in 168 of 172 infants and children in the 30 microg group (97.7%; 95% CI, 94.2%-99.1%). Corresponding seroconversion rates were 86.8% (95% CI, 80.9%-91.0%) and 94.2% (95% CI, 89.6%-96.8%), and factor increases in geometric mean titer were 13.6 (95% CI, 11.8-15.6) and 18.3 (95% CI, 15.7-21.4). All participants demonstrated antibody titers of 1:40 or greater after the second vaccine dose. Immune responses were robust regardless of age, baseline serostatus, or seasonal influenza vaccination status. The majority of adverse events were mild to moderate in severity. CONCLUSION: One 15-microg dose of vaccine was immunogenic in infants and children starting at 6 months of age and vaccine-associated reactions were mild to moderate in severity. TRIAL REGISTRATION: clinicaltrials.gov Identifier: NCT00940108. PMID- 20026599 TI - Applicability of the triad concept for the positional specificity of mammalian lipoxygenases. AB - The nomenclature of lipoxygenases (LOXs) is partly based on the positional specificity of arachidonic acid oxygenation, but there is no unifying concept explaining the mechanistic basis of this enzyme property. According to the triad model, Phe-353, Ile-418, and Ile-593 of the rabbit 12/15-LOX form the bottom of the substrate-binding pocket, and introduction of less space-filling residues at either of these positions favors arachidonic acid 12-lipoxygenation. The present study was aimed at exploring the validity of the triad concept for two novel primate 12/15-LOX (Macaca mulatta and Pongo pygmaeus) and for five known members of the mammalian LOX family (human 12/15-LOX, mouse 12/15-LOX, human 15-LOX2, human platelet type 12-LOX, and mouse (12R)-LOX). The enzymes were expressed as N terminal His tag fusion proteins in E. coli, the potential sequence determinants were mutated, and the specificity of arachidonic acid oxygenation was quantified. Taken together, our data indicate that the triad concept explains the positional specificity of all 12/15-LOXs tested (rabbit, human, M. mulatta, P. pygmaeus, and mouse). For the new enzymes of M. mulatta and P. pygmaeus, the concept had predictive value because the positional specificity predicted on the basis of the amino acid sequence was confirmed experimentally. The specificity of the platelet 12-LOX was partly explained by the triad hypothesis, but the concept was not applicable for 15-LOX2 and (12R)-LOX. PMID- 20026598 TI - The ubiquitin ligase Nedd4-1 is required for heart development and is a suppressor of thrombospondin-1. AB - Nedd4 (Nedd4-1) is a Hect domain E3 ubiquitin ligase that also contains a C2 domain and three WW domains. Despite numerous in vitro studies, its biological function in vivo is not well understood. Here we show that disruption of Nedd4-1 in mice (leaving Nedd4-2 intact) caused embryonic lethality at mid gestation, with pronounced heart defects (double-outlet right ventricle and atrioventricular cushion defects) and vasculature abnormalities. Quantitative mass spectrometry and immunoblot analyses of lysates from the wild type and knock-out mouse embryonic fibroblasts to identify Nedd4-1 in vivo targets revealed dramatically increased amounts of thrombospondin-1 (Tsp-1) in the knock-out mouse embryonic fibroblasts and embryos. Tsp-1 is an inhibitor of angiogenesis, and its elevated level was mediated primarily by enhanced transcription. Interestingly, the administration of aspirin (an inhibitor of Tsp-1) to the pregnant heterozygote mothers led to a reduction in Tsp-1 levels and a substantial rescue of the embryonic lethality. These results suggest that Nedd4-1 is a suppressor of Tsp1 and that increased levels of Tsp-1 in the Nedd4-1 knock-out mice may have contributed to the developmental defect observed in the embryos. PMID- 20026600 TI - Vesicle-associated membrane protein (VAMP) cleavage by a new metalloprotease from the Brazilian scorpion Tityus serrulatus. AB - We present evidence that venom from the Brazilian scorpion Tityus serrulatus and a purified fraction selectively cleave essential SNARE proteins within exocrine pancreatic tissue. Western blotting for vesicle-associated membrane protein type v-SNARE proteins (or synaptobrevins) reveals characteristic alterations to venom treated excised pancreatic lobules in vitro. Immunocytochemistry by electron microscopy confirms both the SNARE identity as VAMP2 and the proteolysis of VAMP2 as a marked decrease in secondary antibody-conjugated colloidal gold particles that are predominantly associated with mature zymogen granules. Studies with recombinant SNARE proteins were used to determine the specific cleavage site in VAMP2 and the susceptibility of VAMP8 (endobrevin). The VAMP2 cleavage site is between the transmembrane anchor and the SNARE motif that assembles into the ternary SNARE complex. Inclusion of divalent chelating agents (EDTA) with fraction nu, an otherwise active purified component from venom, eliminates SNARE proteolysis, suggesting the active protein is a metalloprotease. The unique cleavages of VAMP2 and VAMP8 may be linked to pancreatitis that develops following scorpion envenomation as both of these v-SNARE proteins are associated with zymogen granule membranes in pancreatic acinar cells. We have isolated antarease, a metalloprotease from fraction nu that cleaves VAMP2, and report its amino acid sequence. PMID- 20026601 TI - Constitutive endocytic recycling and protein kinase C-mediated lysosomal degradation control K(ATP) channel surface density. AB - Pancreatic ATP-sensitive potassium (K(ATP)) channels control insulin secretion by coupling the excitability of the pancreatic beta-cell to glucose metabolism. Little is currently known about how the plasma membrane density of these channels is regulated. We therefore set out to examine in detail the endocytosis and recycling of these channels and how these processes are regulated. To achieve this goal, we expressed K(ATP) channels bearing an extracellular hemagglutinin epitope in human embryonic kidney cells and followed their fate along the endocytic pathway. Our results show that K(ATP) channels undergo multiple rounds of endocytosis and recycling. Further, activation of protein kinase C (PKC) with phorbol 12-myristate 13-acetate significantly decreases K(ATP) channel surface density by reducing channel recycling and diverting the channel to lysosomal degradation. These findings were recapitulated in the model pancreatic beta-cell line INS1e, where activation of PKC leads to a decrease in the surface density of native K(ATP) channels. Because sorting of internalized channels between lysosomal and recycling pathways could have opposite effects on the excitability of pancreatic beta-cells, we propose that PKC-regulated K(ATP) channel trafficking may play a role in the regulation of insulin secretion. PMID- 20026602 TI - The ubiquitin binding region of the Smurf HECT domain facilitates polyubiquitylation and binding of ubiquitylated substrates. AB - Mono- and polyubiquitylation of proteins are key steps in a wide range of biological processes. However, the molecular mechanisms that mediate these different events are poorly understood. Here, we employed NMR spectroscopy to map a non-covalent ubiquitin binding surface (UBS) on the Smurf ubiquitin ligase HECT domain. Analysis of mutants of the HECT UBS reveal that interfering with the UBS surface blocked Smurf-dependent degradation of its substrate RhoA in cells. In vitro analysis revealed that the UBS was not required for UbcH7-dependent charging of the HECT catalytic cysteine. Surprisingly, although the UBS was required for polyubiquitylation of both Smurf itself and the Smurf substrate RhoA, it was not required for monoubiquitylation. Furthermore, we show that mutating the UBS interfered with efficient binding of a monoubiquitylated form of RhoA to the Smurf HECT domain. Our findings suggest the UBS promotes polyubiquitylation by stabilizing ubiquitylated substrate binding to the HECT domain. PMID- 20026603 TI - Farnesoid X receptor, through the binding with steroidogenic factor 1-responsive element, inhibits aromatase expression in tumor Leydig cells. AB - The farnesoid X receptor (FXR) is a member of the nuclear receptor superfamily that regulates bile acid homeostasis. It is expressed in the liver and the gastrointestinal tract, but also in several non-enterohepatic tissues including testis. Recently, FXR was identified as a negative modulator of the androgen estrogen-converting aromatase enzyme in human breast cancer cells. In the present study we detected the expression of FXR in Leydig normal and tumor cell lines and in rat testes tissue. We found, in rat Leydig tumor cells, R2C, that FXR activation by the primary bile acid chenodeoxycholic acid (CDCA) or a synthetic agonist GW4064, through a SHP-independent mechanism, down-regulates aromatase expression in terms of mRNA, protein levels, and its enzymatic activity. Transient transfection experiments, using vector containing rat aromatase promoter PII, evidenced that CDCA reduces basal aromatase promoter activity. Mutagenesis studies, electrophoretic mobility shift, and chromatin immunoprecipitation analysis reveal that FXR is able to compete with steroidogenic factor 1 in binding to a common sequence present in the aromatase promoter region interfering negatively with its activity. Finally, the FXR mediated anti-proliferative effects exerted by CDCA on tumor Leydig cells are at least in part due to an inhibition of estrogen-dependent cell growth. In conclusion our findings identify for the first time the activators of FXR as negative modulators of the aromatase enzyme in Leydig tumor cell lines. PMID- 20026604 TI - Intercellular transfer of proteins as identified by stable isotope labeling of amino acids in cell culture. AB - We tracked the extracellular fate of proteins of pulmonary origin using the technique of stable isotope labeling of amino acids in cell culture (SILAC) in cell-impermeable Transwell culture systems. We find that irradiation to murine lung and lung-derived cells induces their release of proteins that are capable of entering neighboring cells, including primary murine bone marrow cells as well as prostate cancer and hematopoietic cell lines. The functional classification of transferred proteins was broad and included transcription factors, mediators of basic cellular processes and components of the nucleosome remodeling and deacetylase complex, including metastasis associated protein 3 and retinoblastoma binding protein 7. In further analysis we find that retinoblastoma-binding protein 7 is a transcriptional activator of E-cadherin and that its intercellular transfer leads to decreased gene expression of downstream targets such as N cadherin and vimentin. SILAC-generated data sets offer a valuable tool to identify and validate potential paracrine networks that may impact relevant biologic processes associated with phenotypic and genotypic signatures of health and disease. PMID- 20026605 TI - Dual specificity of Langerin to sulfated and mannosylated glycans via a single C type carbohydrate recognition domain. AB - Langerin is categorized as a C-type lectin selectively expressed in Langerhans cells, playing roles in the first line of defense against pathogens and in Birbeck granule formation. Although these functions are thought to be exerted through glycan-binding activity of the C-type carbohydrate recognition domain, sugar-binding properties of Langerin have not been fully elucidated in relation to its biological functions. Here, we investigated the glycan-binding specificity of Langerin using comprehensive glycoconjugate microarray, quantitative frontal affinity chromatography, and conventional cell biological analyses. Langerin showed outstanding affinity to galactose-6-sulfated oligosaccharides, including keratan sulfate, while it preserved binding activity to mannose, as a common feature of the C-type lectins with an EPN motif. By a mutagenesis study, Lys-299 and Lys-313 were found to form extended binding sites for sulfated glycans. Consistent with the former observation, the sulfated Langerin ligands were found to be expressed in brain and spleen, where the transcript of keratan sulfate 6-O sulfotransferase is expressed. Moreover, such sulfated ligands were up-regulated in glioblastoma relative to normal brain tissues, and Langerin-expressing cells were localized in malignant brain tissues. Langerin also recognized pathogenic fungi, such as Candida and Malassezia, expressing heavily mannosylated glycans. These observations provide strong evidence that Langerin mediates diverse functions on Langerhans cells through dual recognition of sulfated as well as mannosylated glycans by its uniquely evolved C-type carbohydrate-recognition domain. PMID- 20026606 TI - Leukotriene BLT2 receptor monomers activate the G(i2) GTP-binding protein more efficiently than dimers. AB - Accumulating evidence indicates that G protein-coupled receptors can assemble as dimers/oligomers but the role of this phenomenon in G protein coupling and signaling is not yet clear. We have used the purified leukotriene B(4) receptor BLT2 as a model to investigate the capacity of receptor monomers and dimers to activate the adenylyl cyclase inhibitory G(i2) protein. For this, we overexpressed the recombinant receptor as inclusion bodies in the Escherichia coli prokaryotic system, using a human alpha(5) integrin as a fusion partner. This strategy allowed the BLT2 as well as several other G protein-coupled receptors from different families to be produced and purified in large amounts. The BLT2 receptor was then successfully refolded to its native state, as measured by high-affinity LTB(4) binding in the presence of the purified G protein G alpha(i2). The receptor dimer, in which the two protomers displayed a well defined parallel orientation as assessed by fluorescence resonance energy transfer, was then separated from the monomer. Using two methods of receptor catalyzed guanosine 5'-3-O-(thio)triphosphate binding assay, we clearly demonstrated that monomeric BLT2 stimulates the purified G alpha(i2) beta(1) gamma(2) protein more efficiently than the dimer. These data suggest that assembly of two BLT2 protomers into a dimer results in the reduced ability to signal. PMID- 20026608 TI - A calcium/calmodulin-regulated member of the receptor-like kinase family confers cold tolerance in plants. AB - Cold is a limiting environmental factor that adversely affects plant growth and productivity. Calcium/calmodulin-mediated signaling is believed to play a pivotal role in plant response to cold stress, but its exact role is not clearly understood. Here, we report that CRLK1, a novel calcium/calmodulin-regulated receptor-like kinase, is crucial for cold tolerance in plants. CRLK1 has two calmodulin-binding sites with different affinities as follows: one located at residues 369-390 with a K(d) of 25 nm, and the other located at residues 28-112 with a K(d) of 160 nm. Calcium/calmodulin stimulated the kinase activity, but the addition of chlorpromazine, a calmodulin antagonist, blocked its stimulation. CRLK1 is mainly localized in the plasma membrane, and its expression is stimulated by cold and hydrogen peroxide treatments. Under normal growth conditions, there is no noticeable phenotypic difference between wild-type and crlk1 knock-out mutant plants. However, as compared with wild-type plants, the crlk1 knock-out mutants exhibited an increased sensitivity to chilling and freezing temperatures. Northern analysis showed that the induction of cold responsive genes, including CBF1, RD29A, COR15a, and KIN1 in crlk1 mutants, is delayed as compared with wild-type plants. These results indicate that CRLK1 is a positive regulator of cold tolerance in plants. Furthermore, our results suggest that CRLK1 plays a role in bridging calcium/calmodulin signaling and cold signaling. PMID- 20026607 TI - Repair of O4-alkylthymine by O6-alkylguanine-DNA alkyltransferases. AB - O(6)-Alkylguanine-DNA alkyltransferase (AGT) plays a major role in repair of the cytotoxic and mutagenic lesion O(6)-methylguanine (m(6)G) in DNA. Unlike the Escherichia coli alkyltransferase Ogt that also repairs O(4)-methylthymine (m(4)T) efficiently, the human AGT (hAGT) acts poorly on m(4)T. Here we made several hAGT mutants in which residues near the cysteine acceptor site were replaced by corresponding residues from Ogt to investigate the basis for the inefficiency of hAGT in repair of m(4)T. Construct hAGT-03 (where hAGT sequence V(149)CSSGAVGN(157)- was replaced with the corresponding Ogt -I(143)GRNGTMTG(151) ) exhibited enhanced m(4)T repair activity in vitro compared with hAGT. Three AGT proteins (hAGT, hAGT-03, and Ogt) exhibited similar protection from killing by N methyl-N'-nitro-N-nitrosoguanidine and caused a reduction in m(6)G-induced G:C to A:T mutations in both nucleotide excision repair (NER)-proficient and -deficient Escherichia coli strains that lack endogenous AGTs. hAGT-03 resembled Ogt in totally reducing the m(4)T-induced T:A to C:G mutations in NER-proficient and deficient strains. Surprisingly, wild type hAGT expression caused a significant but incomplete decrease in NER-deficient strains but a slight increase in T:A to C:G mutation frequency in NER-proficient strains. The T:A to C:G mutations due to O(4)-alkylthymine formed by ethylating and propylating agents were also efficiently reduced by either hAGT-03 or Ogt, whereas hAGT had little effect irrespective of NER status. These results show that specific alterations in the hAGT active site facilitate efficient recognition and repair of O(4) alkylthymines and reveal damage-dependent interactions of base and nucleotide excision repair. PMID- 20026610 TI - Risk of cardiovascular disease in HIV-infected patients. AB - The life expectancy of people living with HIV infection has improved dramatically since the use of highly active antiretroviral therapy (HAART). Now that patients with HIV infection are living longer, the focus of its treatment should shift to long-term management spanning decades. Diseases of ageing, including cardiovascular disease (CVD), have now become more important. The evidence of cardiovascular risk associated with HIV infection and antiretroviral therapy is explored and discussed in this article. PMID- 20026609 TI - Dynamic changes in the subcellular distribution of Gpd1p in response to cell stress. AB - Gpd1p is a cytosolic NAD(+)-dependent glycerol 3-phosphate dehydrogenase that also localizes to peroxisomes and plays an essential role in the cellular response to osmotic stress and a role in redox balance. Here, we show that Gpd1p is directed to peroxisomes by virtue of an N-terminal type 2 peroxisomal targeting signal (PTS2) in a Pex7p-dependent manner. Significantly, localization of Gpd1p to peroxisomes is dependent on the metabolic status of cells and the phosphorylation of aminoacyl residues adjacent to the targeting signal. Exposure of cells to osmotic stress induces changes in the subcellular distribution of Gpd1p to the cytosol and nucleus. This behavior is similar to Pnc1p, which is coordinately expressed with Gpd1p, and under conditions of cell stress changes its subcellular distribution from peroxisomes to the nucleus where it mediates chromatin silencing. Although peroxisomes are necessary for the beta-oxidation of fatty acids in yeast, the localization of Gpd1p to peroxisomes is not. Rather, shifts in the distribution of Gpd1p to different cellular compartments in response to changing cellular status suggests a role for Gpd1p in the spatial regulation of redox potential, a process critical to cell survival, especially under the complex stress conditions expected to occur in the wild. PMID- 20026611 TI - Visceral leishmaniasis affects liver and spleen concentrations of amphotericin B following administration to mice. AB - OBJECTIVES: To assess the impact of visceral leishmaniasis (VL) on the concentration of amphotericin B (AmB) recovered in the liver and spleen following either intravenous (AmBisome) or oral (iCo-009) AmB administration to mice. METHODS: Livers and spleens previously obtained from VL-infected BALB/c mice (following intravenous AmBisome or oral AmB treatments) were analysed for AmB concentrations. Then, non-infected BALB/c mice were divided into three treatment groups: a single dose of intravenous AmBisome (2 mg/kg, n = 5); and oral AmB every 12 h for 5 days (10 mg/kg, n = 6 and 20 mg/kg, n = 6). The animals were sacrificed 7 days after the initiation of the treatment and the livers and spleens were harvested for drug analysis by HPLC. RESULTS: The single intravenous injection of AmBisome resulted in a 77-fold lower concentration of AmB in infected compared with non-infected liver tissue, while the difference in AmB concentration in the spleen was only 5-fold. The multiple dose oral administration of AmB resulted in a 3-fold lower concentration of AmB in infected compared with non-infected livers for both oral doses, while the differences in AmB concentrations in the spleen were not statistically different for the oral treatment groups. CONCLUSIONS: VL significantly lowered the concentration of AmB in the liver and the spleen when compared with uninfected animals. This effect seems to correlate with the degree of infection of the tissue. In the case of the intravenous liposomal formulation (AmBisome), the differences between the infected and non-infected tissues are of a higher magnitude than in the case of orally administered AmB (iCo-009). PMID- 20026612 TI - In vitro pharmacodynamic models to determine the effect of antibacterial drugs. AB - In vitro pharmacodynamic (PD) models are used to obtain useful quantitative information on the effect of either single drugs or drug combinations against bacteria. This review provides an overview of in vitro PD models and their experimental implementation. Models are categorized on the basis of whether the drug concentration remains constant or changes and whether there is a loss of bacteria from the system. Further subdifferentiation is based on whether bacterial loss involves dilution of the medium or is associated with dialysis or diffusion. For comprehension of the underlying principles, experimental settings are simplified and schematically illustrated, including the simulations of various in vivo routes of administration. The different model types are categorized and their (dis)advantages discussed. The application of in vitro models to special organs, infections and pathogens is comprehensively presented. Finally, the relevance and perspectives of in vitro investigations in drug discovery and clinical research are elucidated and discussed. PMID- 20026614 TI - Non-linear vector summation of left and right vestibular signals for human balance. AB - The left and right vestibular organs always transduce the same signal of head movement, and with natural stimuli can only be activated simultaneously. To investigate how signals from the left and right vestibular organs are integrated to control human balance we electrically modulated the firing of vestibular afferents from each labyrinth independently and measured the resulting balance responses. Stimulation of one side at a time (monaural) showed that individual leg muscles receive equal inputs from the two labyrinths even though a single labyrinth appeared capable of signalling 3-D head motion. To deduce principles of left-right integration, balance responses to simultaneous stimulation of both sides (binaural) were compared with responses to monaural stimuli. The binaural whole-body response direction was compatible with vector summation of the left and right monaural responses. The binaural response magnitude, however, was only 64-74% that predicted by the monaural sum. This probably reflects a central non linearity between vestibular input and motor output because stimulation of just one labyrinth revealed a power law relationship between stimulus current and response size with exponents 0.56 (force) and 0.51 (displacement). Thus, doubling total signal magnitude either by doubling monaural current or by binaural stimulation produced equivalent responses. We conclude that both labyrinths provide independent estimates of head motion that are summed vectorially and transformed non-linearly into motor output. The former process improves signal-to noise and reduces artifactual common-mode changes, while the latter enhances responses to small signals, all critical for detecting the small head movements needed to control human balance. PMID- 20026615 TI - Ionic flow enhances low-affinity binding: a revised mechanistic view into Mg2+ block of NMDA receptors. AB - The N-methyl-d-aspartate receptor (NMDAR) channel is one of the major excitatory amino acid receptors in the mammalian brain. Since external Mg(2+) blocks the channel in an apparently voltage-dependent fashion, this ligand-gated channel displays intriguing voltage-dependent control of Na(+) and Ca(2+) permeability and thus plays an important role in synaptic physiology. We found that the essential features of Mg(2+) block could not be solely envisaged by binding of a charged blocker in the membrane electric field. Instead, the blocking effect of Mg(2+) is critically regulated by, and quantitatively correlated with, the relative tendency of outward and inward ionic fluxes. The 'intrinsic' affinity of Mg(2+) to the binding sites, however, is low (in the millimolar range) in the absence of net ionic flow at 0 mV. Besides, extracellular and intracellular Mg(2+) blocks the channel at distinct sites of electrical distances 0.7 and 0.95 from the outside, respectively. The two sites are separated by a high energy barrier for the movement of Mg(2+) (but not Na(+) or the other ions), and functionally speaking, each could accommodate 1.1 and 0.8 coexisting permeating ions, respectively. Mg(2+) block of the ionic flow thus is greatly facilitated by the flux-coupling effect or the ionic flow (the preponderant direction of permeant ion movement) per se, as if the poorly permeable Mg(2+) is 'pushed' against a high energy barrier by the otherwise permeating ions. Extracellular and intracellular Mg(2+) block then is in essence 'use dependent', more strongly inhibiting both Na(+) and Ca(2+) fluxes with stronger tendencies of influx and efflux, respectively. In conclusion, although permeant ions themselves could compete with Mg(2+), the flow or the tendency of movement of the permeant ions may actually enhance rather than interfere with Mg(2+) block, making the unique current-voltage relationship of NMDAR and the molecular basis of many important neurobiological phenomena. PMID- 20026616 TI - Subunit-specific desensitization of heteromeric kainate receptors. AB - Kainate receptor subunits can form functional channels as homomers of GluK1, GluK2 or GluK3, or as heteromeric combinations with each other or incorporating GluK4 or GluK5 subunits. However, GluK4 and GluK5 cannot form functional channels by themselves. Incorporation of GluK4 or GluK5 into a heteromeric complex increases glutamate apparent affinity and also enables receptor activation by the agonist AMPA. Utilizing two-electrode voltage clamp of Xenopus oocytes injected with cRNA encoding kainate receptor subunits, we have observed that heteromeric channels composed of GluK2/GluK4 and GluK2/GluK5 have steady state concentration response curves that were bell-shaped in response to either glutamate or AMPA. By contrast, homomeric GluK2 channels exhibited a monophasic steady state concentration-response curve that simply plateaued at high glutamate concentrations. By fitting several specific Markov models to GluK2/GluK4 heteromeric and GluK2 homomeric concentration-response data, we have determined that: (a) two strikingly different agonist binding affinities exist; (b) the high affinity binding site leads to channel opening; and (c) the low-affinity agonist binding site leads to strong desensitization after agonist binding. Model parameters also approximate the onset and recovery kinetics of desensitization observed for macroscopic currents measured from HEK-293 cells expressing GluK2 and GluK4 subunits. The GluK2(E738D) mutation lowers the steady state apparent affinity for glutamate by 9000-fold in comparison to GluK2 homomeric wildtype receptors. When this mutant subunit was expressed with GluK4, the rising phase of the glutamate steady state concentration-response curve overlapped with the wildtype curve, whereas the declining phase was right-shifted toward lower affinity. Taken together, these data are consistent with a scheme whereby high affinity agonist binding to a non-desensitizing GluK4 subunit opens the heteromeric channel, whereas low-affinity agonist binding to GluK2 desensitizes the whole channel complex. PMID- 20026617 TI - Cftr and ENaC ion channels mediate NaCl absorption in the mouse submandibular gland. AB - Cystic fibrosis is caused by mutations in CFTR, the cystic fibrosis transmembrane conductance regulator gene. Disruption of CFTR-mediated anion conductance results in defective fluid and electrolyte movement in the epithelial cells of organs such as the pancreas, airways and sweat glands, but the function of CFTR in salivary glands is unclear. Salivary gland acinar cells produce an isotonic, plasma-like fluid, which is subsequently modified by the ducts to produce a hypotonic, NaCl-depleted final saliva. In the present study we investigated whether submandibular salivary glands (SMGs) in F508 mice (Cftr(F/F)) display ion transport defects characteristic of cystic fibrosis in other tissues. Immunolocalization and whole-cell recordings demonstrated that Cftr and the epithelial Na(+) (ENaC) channels are co-expressed in the apical membrane of submandibular duct cells, consistent with the significantly higher saliva [NaCl] observed in vivo in Cftr(F/F) mice. In contrast, Cftr and ENaC channels were not detected in acinar cells, nor was saliva production affected in Cftr(F/F) mice, implying that Cftr contributes little to the fluid secretion process in the mouse SMG. To identify the source of the NaCl absorption defect in Cftr(F/F) mice, saliva was collected from ex vivo perfused SMGs. Cftr(F/F) glands secreted saliva with significantly increased [NaCl]. Moreover, pharmacological inhibition of either Cftr or ENaC in the ex vivo SMGs mimicked the Cftr(F/F) phenotype. In summary, our results demonstrate that NaCl absorption requires and is likely to be mediated by functionally dependent Cftr and ENaC channels localized to the apical membranes of mouse salivary gland duct cells. PMID- 20026618 TI - Influence of high altitude on cerebrovascular and ventilatory responsiveness to CO2. AB - An altered acid-base balance following ascent to high altitude has been well established. Such changes in pH buffering could potentially account for the observed increase in ventilatory CO(2) sensitivity at high altitude. Likewise, if [H(+)] is the main determinant of cerebrovascular tone, then an alteration in pH buffering may also enhance the cerebral blood flow (CBF) responsiveness to CO(2) (termed cerebrovascular CO(2) reactivity). However, the effect altered acid-base balance associated with high altitude ascent on cerebrovascular and ventilatory responsiveness to CO(2) remains unclear. We measured ventilation , middle cerebral artery velocity (MCAv; index of CBF) and arterial blood gases at sea level and following ascent to 5050 m in 17 healthy participants during modified hyperoxic rebreathing. At 5050 m, resting , MCAv and pH were higher (P < 0.01), while bicarbonate concentration and partial pressures of arterial O(2) and CO(2) were lower (P < 0.01) compared to sea level. Ascent to 5050 m also increased the hypercapnic MCAv CO(2) reactivity (2.9 +/- 1.1 vs. 4.8 +/- 1.4% mmHg(1); P < 0.01) and CO(2) sensitivity (3.6 +/- 2.3 vs. 5.1 +/- 1.7 l min(1) mmHg(1); P < 0.01). Likewise, the hypocapnic MCAv CO(2) reactivity was increased at 5050 m (4.2 +/- 1.0 vs. 2.0 +/- 0.6% mmHg(1); P < 0.01). The hypercapnic MCAv CO(2) reactivity correlated with resting pH at high altitude (R(2) = 0.4; P < 0.01) while the central chemoreflex threshold correlated with bicarbonate concentration (R(2) = 0.7; P < 0.01). These findings indicate that (1) ascent to high altitude increases the ventilatory CO(2) sensitivity and elevates the cerebrovascular responsiveness to hypercapnia and hypocapnia, and (2) alterations in cerebrovascular CO(2) reactivity and central chemoreflex may be partly attributed to an acid-base balance associated with high altitude ascent. Collectively, our findings provide new insights into the influence of high altitude on cerebrovascular function and highlight the potential role of alterations in acid base balance in the regulation in CBF and ventilatory control. PMID- 20026619 TI - Localisation of the mechanotransducer channels in mammalian cochlear hair cells provides clues to their gating. AB - Our sense of hearing and balance relies on the very rapid gating of mechanotransducer channels known to be located close to the tops of the hair cell stereocilia within the stereociliary bundle. The molecular identity of the channels is unknown but functional aspects such as permeation, block and sensitivity to bundle displacement are well known. The channel has high calcium permeability and this feature has been used in conjunction with fast confocal calcium imaging to unambiguously localise the channels at the top of the two shorter rows of stereocilia in mammalian cochlear hair cells. The data suggest that they are completely absent from the tallest row. It is thought that the structures connecting stereocilia in adjacent rows, the tip links, are either directly responsible for the channel's mechanical gating, or are closely associated with the gating process. The channels must therefore be associated with the bottom part of the tip links and not the top. This feature has important implications for both the channel's gating mechanism and its regulatory adaptation mechanism. The tip link remains an attractive candidate for mechanical coupling between the bundle and the channel or an accessory protein. The localisation of the mechanotransducer channels to the lower end of the tip link represents an important milestone in the journey towards eventual identification of the channel and its gating mechanism. PMID- 20026620 TI - Specific expression of an oxytocin-enhanced cyan fluorescent protein fusion transgene in the rat hypothalamus and posterior pituitary. AB - We have generated rats bearing an oxytocin (OXT)-enhanced cyan fluorescent protein (eCFP) fusion transgene designed from a murine construct previously shown to be faithfully expressed in transgenic mice. In situ hybridisation histochemistry revealed that the Oxt-eCfp fusion gene was expressed in the supraoptic nucleus (SON) and the paraventricular nucleus (PVN) in these rats. The fluorescence emanating from eCFP was observed only in the SON, the PVN, the internal layer of the median eminence and the posterior pituitary (PP). In in vitro preparations, freshly dissociated cells from the SON and axon terminals showed clear eCFP fluorescence. Immunohistochemistry for OXT and arginine vasopressin (AVP) revealed that the eCFP fluorescence co-localises with OXT immunofluorescence, but not with AVP immunofluorescence in the SON and the PVN. Although the expression levels of the Oxt-eCfp fusion gene in the SON and the PVN showed a wide range of variations in transgenic rats, eCFP fluorescence was markedly increased in the SON and the PVN, but decreased in the PP after chronic salt loading. The expression of the Oxt gene was significantly increased in the SON and the PVN after chronic salt loading in both non-transgenic and transgenic rats. Compared with wild-type animals, euhydrated and salt-loaded male and female transgenic rats showed no significant differences in plasma osmolality, sodium concentration and OXT and AVP levels, suggesting that the fusion gene expression did not disturb any physiological processes. These results suggest that our new transgenic rats are a valuable new tool to identify OXT-producing neurones and their terminals. PMID- 20026621 TI - Intended transcriptional silencing with siRNA results in gene repression through sequence-specific off-targeting. AB - Transcriptional gene silencing has been reported with siRNA targeting the promoter region of genes. We tested several siRNAs directed against the human VEGF promoter. Of these, siVFp(-992) exhibited > or =50% suppression of VEGF production in two human cell lines. To determine the specificity of this siRNA mediated suppression, plasmids were prepared to express a luciferase reporter under the control of VEGF promoters featuring wild-type, mutated, or deleted target sequences. siRNA transfection assays established sequence-specific inhibition of luciferase from the reporter plasmid featuring the wild-type VEGF promoter. However, siVFp(-992) also suppressed the luciferase expression from the plasmids with mutated or deleted target sites, suggesting that silencing was due to a sequence-specific off-target phenomenon, and this was supported by subsequent microarray and bioinformatics analyses. To determine if our concerns regarding the specificity of promoter targeting siRNAs were relevant to other systems where RNA-mediated transcriptional silencing had been previously reported, we tested a published small RNA sequence directed to the HIV(SF2)-LTR promoter. siRNA transfection assays performed in human cells expressing a luciferase reporter gene under the control of the HIV(SF2)-LTR promoter revealed significant suppression whether the target sequence was intact or mutated, or when the entire HIV(SF2)-LTR was replaced by an irrelevant promoter. These data stress the need to examine target specificity when conducting investigations into transcriptional gene regulation with siRNA. PMID- 20026622 TI - Conserved long noncoding RNAs transcriptionally regulated by Oct4 and Nanog modulate pluripotency in mouse embryonic stem cells. AB - The genetic networks controlling stem cell identity are the focus of intense interest, due to their obvious therapeutic potential as well as exceptional relevance to models of early development. Genome-wide mapping of transcriptional networks in mouse embryonic stem cells (mESCs) reveals that many endogenous noncoding RNA molecules, including long noncoding RNAs (lncRNAs), may play a role in controlling the pluripotent state. We performed a genome-wide screen that combined full-length mESC transcriptome genomic mapping data with chromatin immunoprecipitation genomic location maps of the key mESC transcription factors Oct4 and Nanog. We henceforth identified four mESC-expressed, conserved lncRNA encoding genes residing proximally to active genomic binding sites of Oct4 and Nanog. Accordingly, these four genes have potential roles in pluripotency. We show that two of these lncRNAs, AK028326 (Oct4-activated) and AK141205 (Nanog repressed), are direct targets of Oct4 and Nanog. Most importantly, we demonstrate that these lncRNAs are not merely controlled by mESC transcription factors, but that they themselves regulate developmental state: knockdown and overexpression of these transcripts lead to robust changes in Oct4 and Nanog mRNA levels, in addition to alterations in cellular lineage-specific gene expression and in the pluripotency of mESCs. We further characterize AK028326 as a co activator of Oct4 in a regulatory feedback loop. These results for the first time implicate lncRNAs in the modulation of mESC pluripotency and expand the established mESC regulatory network model to include functional lncRNAs directly controlled by key mESC transcription factors. PMID- 20026623 TI - The obesity epidemic: the role of addiction. PMID- 20026624 TI - Blood glucose test strips: options to reduce usage. AB - BACKGROUND: Recent evidence suggests that, despite widespread use, self monitoring of blood glucose levels has little clinical benefit in many patients with diabetes. The impact of more focused public-payer policies for the use of blood glucose test strips may be substantial. METHODS: We conducted a cross sectional analysis of annual prescription claims for test strips between 1997 and 2008 for patients in Ontario aged 65 and older with diabetes. Patients were stratified into 1 of 4 hierarchical groups according to the most intensive glucose-lowering treatment received during each calendar year. Test strip use was calculated annually for each group over the study period, and the effects of 5 hypothetical policy scenarios of more selective test strip use were assessed. RESULTS: Test strip use increased by almost 250% from 1997 to 2008, with 52.6% (n = 263,513) of included patients receiving a prescription during 2008. Almost half of these patients were at low risk for drug-induced hypoglycemia. In 2008, over 117 million test strips were dispensed in Ontario; however, more focused policy scenarios could have reduced this number by between 9.5 million and 74.5 million test strips. INTERPRETATION: Many people who self-monitor their blood glucose are at relatively low risk for drug-induced hypoglycemia. The economic benefits associated with more selective testing could be redirected to more effective interventions for patients with diabetes. PMID- 20026625 TI - The Goudge Inquiry and the role of medical expert witnesses. PMID- 20026626 TI - Cost-effectiveness of self-monitoring of blood glucose in patients with type 2 diabetes mellitus managed without insulin. AB - BACKGROUND: The benefits of self-monitoring blood glucose levels are unclear in patients with type 2 diabetes mellitus who do not use insulin, but there are considerable costs. We sought to determine the cost effectiveness of self monitoring for patients with type 2 diabetes not using insulin. METHODS: We performed an incremental cost-effectiveness analysis of the self-monitoring of blood glucose in adults with type 2 diabetes not taking insulin. We used the United Kingdom Prospective Diabetes Study (UKPDS) model to forecast diabetes related complications, corresponding quality-adjusted life years and costs. Clinical data were obtained from a systematic review comparing self-monitoring with no self-monitoring. Costs and utility decrements were derived from published sources. We performed sensitivity analyses to examine the robustness of the results. RESULTS: Based on a clinically modest reduction in hemoglobin A(1C) of 0.25% (95% confidence interval 0.15-0.36) estimated from the systematic review, the UKPDS model predicted that self-monitoring performed 7 or more times per week reduced the lifetime incidence of diabetes-related complications compared with no self-monitoring, albeit at a higher cost (incremental cost per quality-adjusted life year $113,643). The results were largely unchanged in the sensitivity analysis, although the incremental cost per quality-adjusted life year fell within widely cited cost-effectiveness thresholds when testing frequency or the price per test strip was substantially reduced from the current levels. INTERPRETATION: For most patients with type 2 diabetes not using insulin, use of blood glucose test strips for frequent self-monitoring (>or= 7 times per week) is unlikely to represent efficient use of finite health care resources, although periodic testing (e.g., 1 or 2 times per week) may be cost-effective. Reduced test strip price would likely also improve cost-effectiveness. PMID- 20026627 TI - Herniation through the broad ligament. PMID- 20026628 TI - Second neoplasm: 31-year latency after childhood leukemia. PMID- 20026629 TI - Income-related differences in mortality among people with diabetes mellitus. AB - BACKGROUND: Mortality has declined substantially among people with diabetes mellitus over the last decade. Whether all income groups have benefited equally, however, is unclear. We examined the impact of income on mortality trends among people with diabetes. METHODS: In this population-based, retrospective cohort study, we compared changes in mortality from Apr. 1, 1994, to Mar. 31, 2005, by neighbourhood income strata among people with diabetes aged 30 years or more in the province of Ontario, Canada. RESULTS: Overall, the annual age- and sex adjusted mortality declined, from 4.05% in 1994/95 (95% confidence interval [CI] 3.98%-4.11%) to 2.69% in 2005/06 (95% CI 2.66%-2.73%). The decrease was significantly greater in the highest income group (by 36%) than in the lowest income group (by 31%; p < 0.001). This trend was most pronounced in the younger group (age 30-64 years): the mortality rate ratio widened by more than 40% between the lowest and highest income groups, from 1.12 to 1.59 among women and from 1.14 to 1.60 among men. Income had a much smaller effect on mortality trends in the older group, whose drug costs are subsidized: the income-related difference rose by only 0.9% over the study period. INTERPRETATION: Mortality declined overall among people with diabetes from 1994 to 2005; however, the decrease was substantially greater in the highest income group than in the lowest, particularly among those aged 30-64 years. These findings illustrate the increasing impact of income on the health of people with diabetes even in a publicly funded health care setting. Further studies are needed to explore factors responsible for these income-related differences in mortality. PMID- 20026631 TI - Radiation recall dermatitis. PMID- 20026632 TI - Work-up for Cushing syndrome. PMID- 20026634 TI - England hikes educational requirements for nurses. PMID- 20026633 TI - Developing and selecting interventions for translating knowledge to action. PMID- 20026635 TI - Concern raised over control of cost-benefit research in United States. PMID- 20026636 TI - WHO reports progress in malaria control. PMID- 20026637 TI - Allegations prompt Manitoba pathology probes. PMID- 20026638 TI - In the street. PMID- 20026639 TI - Medicinal maggots cross border at a crawl. PMID- 20026640 TI - Rapid neurite outgrowth in neurosecretory cells and neurons is sustained by the exocytosis of a cytoplasmic organelle, the enlargeosome. AB - Neurite outgrowth is known as a slow (days) process occurring in nerve cells and neurons during neurotrophin treatment and upon transfer to culture, respectively. Using Y27632, a drug that induces activation of Rac1, a downstream step of the neurotrophin signaling cascade, we have identified a new form of outgrowth, which is rapid (<1 hour) and extensive (>500 microm(2) surface enlargement/single cell/first hour). However, this outgrowth takes place only in cells (PC12-27 and SH-SY5Y cells, and embryonic and neonatal neurons) rich in an exocytic organelle, the enlargeosome. Golgi vesicles, TGN vesicles and endosomes are not involved. The need for enlargeosomes for plasma-membrane expansion was confirmed by the appearance of their marker, Ahnak, at the cell surface and by the dependence of neurite outgrowth on VAMP4, the vSNARE of enlargeosome exocytosis. In enlargeosome-rich cells, VAMP4 downregulation also attenuated the slow outgrowth induced by nerve growth factor (NGF). Similar to NGF-induced neurite outgrowth in enlargeosome-lacking cells, the new, rapid, Y27632-induced process required microtubules. Other properties of neurite outgrowth in cells lacking enlargeosomes - such as dependence on VAMP7, on microfilaments, on gene transcription and on protein synthesis, and blockade of mitoses and accumulation of neuronal markers - were not evident. The enlargeosome-sustained process might be useful for the rapid neurite outgrowth at peculiar stages and/or conditions of nerve and neuronal cells. However, its properties and its physiological and pathological role remain to be investigated. PMID- 20026641 TI - Identification of beta-catenin as a target of the intracellular tyrosine kinase PTK6. AB - Disruption of the gene encoding protein tyrosine kinase 6 (PTK6) leads to increased growth, impaired enterocyte differentiation and higher levels of nuclear beta-catenin in the mouse small intestine. Here, we demonstrate that PTK6 associates with nuclear and cytoplasmic beta-catenin and inhibits beta-catenin- and T-cell factor (TCF)-mediated transcription. PTK6 directly phosphorylates beta catenin on Tyr64, Tyr142, Tyr331 and/or Tyr333, with the predominant site being Tyr64. However, mutation of these sites does not abrogate the ability of PTK6 to inhibit beta-catenin transcriptional activity. Outcomes of PTK6-mediated regulation appear to be dependent on its intracellular localization. In the SW620 colorectal adenocarcinoma cell line, nuclear-targeted PTK6 negatively regulates endogenous beta-catenin/TCF transcriptional activity, whereas membrane-targeted PTK6 enhances beta-catenin/TCF regulated transcription. Levels of TCF4 and the transcriptional co-repressor TLE/Groucho increase in SW620 cells expressing nuclear-targeted PTK6. Knockdown of PTK6 in SW620 cells leads to increased beta catenin/TCF transcriptional activity and increased expression of beta-catenin/TCF target genes Myc and Survivin. Ptk6-null BAT-GAL mice, containing a beta-catenin activated LacZ reporter transgene, have increased levels of beta-galactosidase expression in the gastrointestinal tract. The ability of PTK6 to negatively regulate beta-catenin/TCF transcription by modulating levels of TCF4 and TLE/Groucho could contribute to its growth-inhibitory activities in vivo. PMID- 20026642 TI - Persistence of the cell-cycle checkpoint kinase Wee1 in SadA- and SadB-deficient neurons disrupts neuronal polarity. AB - Wee1 is well characterized as a cell-cycle checkpoint kinase that regulates the entry into mitosis in dividing cells. Here we identify a novel function of Wee1 in postmitotic neurons during the establishment of distinct axonal and dendritic compartments, which is an essential step during neuronal development. Wee1 is expressed in unpolarized neurons but is downregulated after neurons have extended an axon. Suppression of Wee1 impairs the formation of minor neurites but does not interfere with axon formation. However, neuronal polarity is disrupted when neurons fail to downregulate Wee1. The kinases SadA and SadB (Sad kinases) phosphorylate Wee1 and are required to initiate its downregulation in polarized neurons. Wee1 expression persists in neurons that are deficient in SadA and SadB and disrupts neuronal polarity. Knockdown of Wee1 rescues the Sada(-/-);Sadb(-/-) mutant phenotype and restores normal polarity in these neurons. Our results demonstrate that the regulation of Wee1 by SadA and SadB kinases is essential for the differentiation of polarized neurons. PMID- 20026643 TI - Interaction with surrounding normal epithelial cells influences signalling pathways and behaviour of Src-transformed cells. AB - At the initial stage of carcinogenesis, transformation occurs in a single cell within an epithelial sheet. However, it remains unknown what happens at the boundary between normal and transformed cells. Using Madin-Darby canine kidney (MDCK) cells transformed with temperature-sensitive v-Src, we have examined the interface between normal and Src-transformed epithelial cells. We show that Src transformed cells are apically extruded when surrounded by normal cells, but not when Src cells alone are cultured, suggesting that apical extrusion occurs in a cell-context-dependent manner. We also observe apical extrusion of Src transformed cells in the enveloping layer of zebrafish gastrula embryos. When Src transformed MDCK cells are surrounded by normal MDCK cells, myosin-II and focal adhesion kinase (FAK) are activated in Src cells, which further activate downstream mitogen-activated protein kinase (MAPK). Importantly, activation of these signalling pathways depends on the presence of surrounding normal cells and plays a crucial role in apical extrusion of Src cells. Collectively, these results indicate that interaction with surrounding normal epithelial cells influences the signalling pathways and behaviour of Src-transformed cells. PMID- 20026644 TI - Casein-kinase-II-dependent phosphorylation of PPARgamma provokes CRM1-mediated shuttling of PPARgamma from the nucleus to the cytosol. AB - PPARgamma exerts significant anti-inflammatory signaling properties in monocytes and macrophages, which are affected by its intracellular localization. Based on our previous report, which showed that cytosolic localization of PPARgamma attenuates PKCalpha signaling in macrophages, we elucidated the molecular mechanisms provoking cytosolic PPARgamma localization. Using the DsRed-tagged PPARgamma deletion constructs PPARgamma1 Delta1-31 and PPARgamma1 Delta407-475, we observed an exclusive nuclear PPARgamma1 Delta1-31 localization in transfected HEK293 cells, whereas PPARgamma1 Delta407-475 did not alter its cytosolic or nuclear localization. The casein kinase II (CK-II) inhibitor 5,6-dichloro-1-beta D-ribofuranosyl benzimidazole (DRB) prevented cytosolic PPARgamma localization. Mutation of two possible CK-II phosphorylation sites at serine 16 and serine 21 of PPARgamma into alanine (PPARgamma S16A/S21A) inhibited cytosolic PPARgamma localization. Moreover, a PPARgamma S16E/S21E mutant that mimicks constitutive phosphorylation of residues 16 and 21, predominantly resides in the cytosol. The CRM1 inhibitor leptomycin B abolished cytosolic PPARgamma localization, suggesting that this is a CRM1-dependent export process. CRM1-mediated PPARgamma export requires Ran and phosphorylated RanBP3. Finally, co-immunoprecipitation studies demonstrated that DRB blocks PPARgamma binding to CRM1, whereas PD98059 inhibits RanBP3 binding to CRM1 and concomitant shuttling from nucleus to cytosol, but does not alter PPARgamma binding to CRM1. We conclude that CK-II dependent PPARgamma phosphorylation at Ser16 and Ser21 is necessary for CRM1/Ran/RanBP3-mediated nucleocytoplasmic translocation of PPARgamma. PMID- 20026645 TI - Rab11-FIP3 links the Rab11 GTPase and cytoplasmic dynein to mediate transport to the endosomal-recycling compartment. AB - Several protein families control intracellular transport processes in eukaryotic cells. Here, we show that the Rab11 GTPase effector protein Rab11-FIP3 (henceforth, FIP3) directly interacts with the dynein light intermediate chain 1 (DLIC-1, gene symbol DYNC1LI1) subunit of the cytoplasmic dynein 1 motor protein complex. We show that Rab11a, FIP3 and DLIC-1 form a ternary complex and that DLIC-1 colocalises with endogenous FIP3 and Rab11a in A431 cells. We demonstrate that association between FIP3 and DLIC-1 at the cell periphery precedes minus-end directed microtubule-based transport, that FIP3 recruits DLIC-1 onto membranes, and that knockdown of DLIC-1 inhibits pericentrosomal accumulation of key endosomal-recycling compartment (ERC) proteins. In addition, we demonstrate that expression of a DLIC-1-binding truncation mutant of FIP3 disrupts the ability of ERC proteins to accumulate pericentrosomally. On the basis of these and other data, we propose that FIP3 links the Rab11 GTPase and cytoplasmic dynein to mediate transport of material from peripheral sorting endosomes to the centrally located ERC. PMID- 20026646 TI - CDC73/HRPT2 CpG island hypermethylation and mutation of 5'-untranslated sequence are uncommon mechanisms of silencing parafibromin in parathyroid tumors. AB - The tumor suppressor HRPT2/CDC73 is mutated in constitutive DNA from patients with the familial disorder hyperparathyroidism-jaw tumor syndrome and in approximately 70% of all parathyroid carcinomas. In a number of HRPT2 mutant tumors however, expression of the encoded protein parafibromin is lost in the absence of a clear second event such as HRPT2 allelic loss or the presence of a second mutation in this tumor suppressor gene. We sought to determine whether hypermethylation of a 713 bp CpG island extending 648 nucleotides upstream of the HRPT2 translational start site and 65 nucleotides into exon 1 might be a mechanism contributing to the loss of expression of parafibromin in parathyroid tumors. Furthermore, we asked whether mutations might be present in the 5' untranslated region (5'-UTR) of HRPT2. We investigated a pool of tissue from 3 normal parathyroid glands, as well as 15 individual parathyroid tumor samples including 6 tumors with known HRPT2 mutations, for hypermethylation of the HRPT2 CpG island. Methylation was not identified in any specimens despite complete loss of parafibromin expression in two parathyroid carcinomas with a single detectable HRPT2 mutation and retention of the wild-type HRPT2 allele. Furthermore, no mutations of a likely pathogenic nature were identified in the 5'-UTR of HRPT2. These data strongly suggest that alternative mechanisms such as mutation in HRPT2 intronic regions, additional epigenetic regulation such as histone modifications, or other regulatory inactivation mechanisms such as targeting by microRNAs may play a role in the loss of parafibromin expression. PMID- 20026647 TI - The long-term survival in adrenocortical carcinoma with active surgical management and use of monitored mitotane. AB - Adrenocortical carcinoma (ACC) is a rare tumour disease with sinister prognosis also after attempts to radical surgery; better prognosis is seen for low-stage tumours. Adjuvant treatment with the adrenolytic drug mitotane has been attempted, but not proven to prevent from recurrence. The drug may offer survival advantage in case of recurrence. The aim of this single-centre study (1979-2007) of 43 consecutive patients was to evaluate the long-term survival after active surgical treatment combined with monitored mitotane (to reduce side effects of the drug). The series is unique, since all patients were offered a period of mitotane as adjuvant or palliative treatment; six patients refused mitotane. Despite a high proportion of high-stage tumours (67%), the complete resection rate was high (77%). The disease-specific 5-year survival was high (64.1%); very high for patients with low-stage tumours without evident relation to mitotane levels. Patients with high-stage tumours had a clear survival advantage with mitotane levels above a threshold of 14 mg/l in serum. The hazard ratio for patients with high mitotane levels versus all patients indicates a significant effect of the drug. The results indicate that adjuvant mitotane may be the standard of care for patients with high-stage ACC after complete resection. PMID- 20026648 TI - The quest for the initial lesion in idiopathic pulmonary fibrosis: gene expression differences in IPF fibroblasts. PMID- 20026649 TI - Erratum: figure correction. PMID- 20026650 TI - Pediatric nurses' perceptions of obstacles and supportive behaviors in end-of life care. AB - BACKGROUND: Each year 55 000 children die in the United States, and most of these deaths occur in hospitals. The barriers and supportive behaviors in providing end of-life care to children should be determined. OBJECTIVE: To determine pediatric intensive care unit nurses' perceptions of sizes, frequencies, and magnitudes of selected obstacles and helpful behaviors in providing end-of-life care to children. METHOD: A national sample of 1047 pediatric intensive care unit nurses who were members of the American Association of Critical-Care Nurses were surveyed. A 76-item questionnaire adapted from 3 similar surveys with critical care, emergency, and oncology nurses was mailed to possible participants. Nurses who did not respond to the first mailing were sent a second mailing. Nurses were asked to rate the size and frequency of listed obstacles and supportive behaviors in caring for children at the end of life. RESULTS: A total of 474 usable questionnaires were received from 985 eligible respondents (return rate, 48%). The 2 items with the highest perceived obstacle magnitude scores for size and frequency means were language barriers and parental discomfort in withholding and/or withdrawing mechanical ventilation. The highest supportive behavior item was allowing time alone with the child when he or she has died. CONCLUSIONS: Pediatric intensive care unit nurses play a vital role in caring for dying children and the children's families. Overcoming language and communication barriers with children's families and between interdisciplinary team members could greatly improve the end-of-life experience for dying children. PMID- 20026651 TI - Current practices for withdrawal of life support in intensive care units. AB - BACKGROUND: Nurses are present at the bedside of patients undergoing withdrawal of life support more often than any other member of the health care team, yet most publications on this topic are directed at physicians. OBJECTIVES: To describe the training, guidance, and support related to withdrawal of life support received by nurses in intensive care units in the United States, how the nurses participated, and how the withdrawal of life support occurred. METHODS: A questionnaire about withdrawal of life support was sent to 1000 randomly selected members of the American Association of Critical-Care Nurses, with 2 follow-up mailings. RESULTS: Responses were received from 48.4% of the nurses surveyed. Content on withdrawal of life support was required in only 15.5% of respondents' basic nursing education and was absent from work site orientations for 63.1% of respondents. Nurses' actions during withdrawal were most often guided by individual physician's orders (63.8%), followed by standardized care plans (20%) and standing orders (11.8%). Nurses rated the importance of emotional support during and after the withdrawal of life support very highly, but they did not believe they were receiving that level of support. Most respondents (87.5%) participated in family conferences where withdrawal of life support was discussed. After physicians, nurses were most influential concerning administration of palliative medications. Patients' families were present during withdrawal procedures between 32.3% and 58.4% of the time. CONCLUSIONS: To improve their practice, intensive care nurses should receive formal training on withdrawal of life support, and institutions should develop best practices that support nurses in providing the highest quality care for patients undergoing this procedure. PMID- 20026652 TI - A novel intermembrane space-targeting signal docks cysteines onto Mia40 during mitochondrial oxidative folding. AB - Mia40 imports Cys-containing proteins into the mitochondrial intermembrane space (IMS) by ensuring their Cys-dependent oxidative folding. In this study, we show that the specific Cys of the substrate involved in docking with Mia40 is substrate dependent, the process being guided by an IMS-targeting signal (ITS) present in Mia40 substrates. The ITS is a 9-aa internal peptide that (a) is upstream or downstream of the docking Cys, (b) is sufficient for crossing the outer membrane and for targeting nonmitochondrial proteins, (c) forms an amphipathic helix with crucial hydrophobic residues on the side of the docking Cys and dispensable charged residues on the other side, and (d) fits complementary to the substrate cleft of Mia40 via hydrophobic interactions of micromolar affinity. We rationalize the dual function of Mia40 as a receptor and an oxidase in a two step-specific mechanism: an ITS-guided sliding step orients the substrate noncovalently, followed by docking of the substrate Cys now juxtaposed to pair with the Mia40 active Cys. PMID- 20026653 TI - The p38 MAPK family, a pushmi-pullyu of skeletal muscle differentiation. AB - In this issue, Gillespie et al. (Gillespie et al. 2009. J. Cell Biol. doi:10.1083/jcb.200907037) demonstrate that the mitogen-activated protein kinase isoform p38-gamma plays a crucial role in blocking the premature differentiation of satellite cells, a skeletal muscle stem cell population. p38-gamma puts the brakes on skeletal muscle differentiation by promoting the association of the transcription factor MyoD with the histone methyltransferase, KMT1A, which act together in a complex to repress the premature expression of the gene encoding the myogenic transcription factor Myogenin. PMID- 20026654 TI - Autophosphorylation at serine 1981 stabilizes ATM at DNA damage sites. AB - Ataxia telangiectasia mutated (ATM) plays a critical role in the cellular response to DNA damage. In response to DNA double-strand breaks (DSBs), ATM is autophosphorylated at serine 1981. Although this autophosphorylation is widely considered a sign of ATM activation, it is still not clear if autophosphorylation is required for ATM functions including localization to DSBs and activation of ATM kinase activity. In this study, we show that localization of ATM to DSBs is differentially regulated with the initial localization requiring the MRE11-RAD50 NBS1 complex and sustained retention requiring autophosphorylation of ATM at serine 1981. Autophosphorylated ATM interacts with MDC1 and the latter is required for the prolonged association of ATM to DSBs. Ablation of ATM autophosphorylation or knock-down of MDC1 protein affects the ability of ATM to phosphorylate downstream substrates and confer radioresistance. Together, these data suggest that autophosphorylation at serine 1981 stabilizes ATM at the sites of DSBs, and this is required for a proper DNA damage response. PMID- 20026655 TI - Actin remodeling by ADF/cofilin is required for cargo sorting at the trans-Golgi network. AB - Knockdown of the actin-severing protein actin-depolymerizing factor (ADF)/cofilin inhibited export of an exogenously expressed soluble secretory protein from Golgi membranes in Drosophila melanogaster and mammalian tissue culture cells. A stable isotope labeling by amino acids in cell culture mass spectrometry-based protein profiling revealed that a large number of endogenous secretory proteins in mammalian cells were not secreted upon ADF/cofilin knockdown. Although many secretory proteins were retained, a Golgi-resident protein and a lysosomal hydrolase were aberrantly secreted upon ADF/cofilin knockdown. Overall, our findings indicate that inactivation of ADF/cofilin perturbed the sorting of a subset of both soluble and integral membrane proteins at the trans-Golgi network (TGN). We suggest that ADF/cofilin-dependent actin trimming generates a sorting domain at the TGN, which filters secretory cargo for export, and that uncontrolled growth of this domain causes missorting of proteins. This type of actin-dependent compartmentalization and filtering of secretory cargo at the TGN by ADF/cofilin could explain sorting of proteins that are destined to the cell surface. PMID- 20026656 TI - IKK phosphorylates Huntingtin and targets it for degradation by the proteasome and lysosome. AB - Expansion of the polyglutamine repeat within the protein Huntingtin (Htt) causes Huntington's disease, a neurodegenerative disease associated with aging and the accumulation of mutant Htt in diseased neurons. Understanding the mechanisms that influence Htt cellular degradation may target treatments designed to activate mutant Htt clearance pathways. We find that Htt is phosphorylated by the inflammatory kinase IKK, enhancing its normal clearance by the proteasome and lysosome. Phosphorylation of Htt regulates additional post-translational modifications, including Htt ubiquitination, SUMOylation, and acetylation, and increases Htt nuclear localization, cleavage, and clearance mediated by lysosomal associated membrane protein 2A and Hsc70. We propose that IKK activates mutant Htt clearance until an age-related loss of proteasome/lysosome function promotes accumulation of toxic post-translationally modified mutant Htt. Thus, IKK activation may modulate mutant Htt neurotoxicity depending on the cell's ability to degrade the modified species. PMID- 20026657 TI - p38-{gamma}-dependent gene silencing restricts entry into the myogenic differentiation program. AB - The mitogen-activated protein kinase p38-gamma is highly expressed in skeletal muscle and is associated with the dystrophin glycoprotein complex; however, its function remains unclear. After induced damage, muscle in mice lacking p38-gamma generated significantly fewer myofibers than wild-type muscle. Notably, p38-gamma deficient muscle contained 50% fewer satellite cells that exhibited premature Myogenin expression and markedly reduced proliferation. We determined that p38 gamma directly phosphorylated MyoD on Ser199 and Ser200, which results in enhanced occupancy of MyoD on the promoter of myogenin together with markedly decreased transcriptional activity. This repression is associated with extensive methylation of histone H3K9 together with recruitment of the KMT1A methyltransferase to the myogenin promoter. Notably, a MyoD S199A/S200A mutant exhibits markedly reduced binding to KMT1A. Therefore, p38-gamma signaling directly induces the assembly of a repressive MyoD transcriptional complex. Together, these results establish a hitherto unappreciated and essential role for p38-gamma signaling in positively regulating the expansion of transient amplifying myogenic precursor cells during muscle growth and regeneration. PMID- 20026658 TI - PtdIns4P recognition by Vps74/GOLPH3 links PtdIns 4-kinase signaling to retrograde Golgi trafficking. AB - Targeting and retention of resident integral membrane proteins of the Golgi apparatus underly the function of the Golgi in glycoprotein and glycolipid processing and sorting. In yeast, steady-state Golgi localization of multiple mannosyltransferases requires recognition of their cytosolic domains by the peripheral Golgi membrane protein Vps74, an orthologue of human GOLPH3/GPP34/GMx33/MIDAS (mitochondrial DNA absence sensitive factor). We show that targeting of Vps74 and GOLPH3 to the Golgi apparatus requires ongoing synthesis of phosphatidylinositol (PtdIns) 4-phosphate (PtdIns4P) by the Pik1 PtdIns 4-kinase and that modulation of the levels and cellular location of PtdIns4P leads to mislocalization of these proteins. Vps74 and GOLPH3 bind specifically to PtdIns4P, and a sulfate ion in a crystal structure of GOLPH3 indicates a possible phosphoinositide-binding site that is conserved in Vps74. Alterations in this site abolish phosphoinositide binding in vitro and Vps74 function in vivo. These results implicate Pik1 signaling in retention of Golgi resident proteins via Vps74 and show that GOLPH3 family proteins are effectors of Golgi PtdIns 4-kinases. PMID- 20026661 TI - Lymphatic endothelial cell sphingosine kinase activity is required for lymphocyte egress and lymphatic patterning. AB - Lymphocyte egress from lymph nodes (LNs) is dependent on sphingosine-1-phosphate (S1P), but the cellular source of this S1P is not defined. We generated mice that expressed Cre from the lymphatic vessel endothelial hyaluronan receptor 1 (Lyve 1) locus and that showed efficient recombination of loxP-flanked genes in lymphatic endothelium. We report that mice with Lyve-1 CRE-mediated ablation of sphingosine kinase (Sphk) 1 and lacking Sphk2 have a loss of S1P in lymph while maintaining normal plasma S1P. In Lyve-1 Cre+ Sphk-deficient mice, lymphocyte egress from LNs and Peyer's patches is blocked. Treatment with pertussis toxin to overcome Galphai-mediated retention signals restores lymphocyte egress. Furthermore, in the absence of lymphatic Sphks, the initial lymphatic vessels in nonlymphoid tissues show an irregular morphology and a less organized vascular endothelial cadherin distribution at cell-cell junctions. Our data provide evidence that lymphatic endothelial cells are an in vivo source of S1P required for lymphocyte egress from LNs and Peyer's patches, and suggest a role for S1P in lymphatic vessel maturation. PMID- 20026662 TI - VEGF-A expression by HSV-1-infected cells drives corneal lymphangiogenesis. AB - Inflammatory lymphangiogenesis plays a crucial role in the development of inflammation and transplant rejection. The mechanisms of inflammatory lymphangiogenesis during bacterial infection, toll-like receptor ligand administration, and wound healing are well characterized and depend on ligands for the vascular endothelial grow factor receptor (VEGFR) 3 that are produced by infiltrating macrophages. But inflammatory lymphangiogenesis in nonlymphoid tissues during chronic viral infection is unstudied. Herpes simplex virus 1 (HSV 1) infection of the cornea is a leading cause of blindness and depends on aberrant host immune responses to antigen within the normally immunologically privileged cornea. We report that corneal HSV-1 infection drives lymphangiogenesis and that corneal lymphatics persist past the resolution of infection. The mechanism of HSV-1-induced lymphangiogenesis was distinct from the described mechanisms of inflammatory lymphangiogenesis. HSV-1-elicited lymphangiogenesis was strictly dependent on VEGF-A/VEGFR-2 signaling but not on VEGFR-3 ligands. Macrophages played no role in the induction of lymphangiogenesis and were not a detectable source of VEGF-A. Rather, using VEGF-A reporter transgenic mice, we have identified infected epithelial cells as the primary source of VEGF-A during HSV-1 infection. Our results indicate that HSV-1 directly induces vascularization of the cornea through up-regulation of VEGF-A expression. PMID- 20026663 TI - Exaggerated inflammation, impaired host defense, and neuropathology in progranulin-deficient mice. AB - Progranulin (PGRN) is a widely expressed protein involved in diverse biological processes. Haploinsufficiency of PGRN in the human causes tau-negative, ubiquitin positive frontotemporal dementia (FTD). However, the mechanisms are unknown. To explore the role of PGRN in vivo, we generated PGRN-deficient mice. Macrophages from these mice released less interleukin-10 and more inflammatory cytokines than wild type (WT) when exposed to bacterial lipopolysaccharide. PGRN-deficient mice failed to clear Listeria monocytogenes infection as quickly as WT and allowed bacteria to proliferate in the brain, with correspondingly greater inflammation than in WT. PGRN-deficient macrophages and microglia were cytotoxic to hippocampal cells in vitro, and PGRN-deficient hippocampal slices were hypersusceptible to deprivation of oxygen and glucose. With age, brains of PGRN deficient mice displayed greater activation of microglia and astrocytes than WT, and their hippocampal and thalamic neurons accumulated cytosolic phosphorylated transactivation response element DNA binding protein-43. Thus, PGRN is a key regulator of inflammation and plays critical roles in both host defense and neuronal integrity. FTD associated with PGRN insufficiency may result from many years of reduced neutrotrophic support together with cumulative damage in association with dysregulated inflammation. PMID- 20026665 TI - Gesundheit! Patrick Holt smothers allergies and asthma. PMID- 20026664 TI - VSOP/Hv1 proton channels sustain calcium entry, neutrophil migration, and superoxide production by limiting cell depolarization and acidification. AB - Neutrophils kill microbes with reactive oxygen species generated by the NADPH oxidase, an enzyme which moves electrons across membranes. Voltage-gated proton channels (voltage-sensing domain only protein [VSOP]/Hv1) are required for high level superoxide production by phagocytes, but the mechanism of this effect is not established. We show that neutrophils from VSOP/Hv1-/- mice lack proton currents but have normal electron currents, indicating that these cells have a fully functional oxidase that cannot conduct protons. VSOP/Hv1-/- neutrophils had a more acidic cytosol, were more depolarized, and produced less superoxide and hydrogen peroxide than neutrophils from wild-type mice. Hydrogen peroxide production was rescued by providing an artificial conductance with gramicidin. Loss of VSOP/Hv1 also aborted calcium responses to chemoattractants, increased neutrophil spreading, and decreased neutrophil migration. The migration defect was restored by the addition of a calcium ionophore. Our findings indicate that proton channels extrude the acid and compensate the charge generated by the oxidase, thereby sustaining calcium entry signals that control the adhesion and motility of neutrophils. Loss of proton channels thus aborts superoxide production and causes a severe signaling defect in neutrophils. PMID- 20026666 TI - Mutation of conserved cysteines in the Ly6 domain of GPIHBP1 in familial chylomicronemia. AB - We investigated a family from northern Sweden in which three of four siblings have congenital chylomicronemia. LPL activity and mass in pre- and postheparin plasma were low, and LPL release into plasma after heparin injection was delayed. LPL activity and mass in adipose tissue biopsies appeared normal. [(35)S]Methionine incorporation studies on adipose tissue showed that newly synthesized LPL was normal in size and normally glycosylated. Breast milk from the affected female subjects contained normal to elevated LPL mass and activity levels. The milk had a lower than normal milk lipid content, and the fatty acid composition was compatible with the milk lipids being derived from de novo lipogenesis, rather than from the plasma lipoproteins. Given the delayed release of LPL into the plasma after heparin, we suspected that the chylomicronemia might be caused by mutations in GPIHBP1. Indeed, all three affected siblings were compound heterozygotes for missense mutations involving highly conserved cysteines in the Ly6 domain of GPIHBP1 (C65S and C68G). The mutant GPIHBP1 proteins reached the surface of transfected Chinese hamster ovary cells but were defective in their ability to bind LPL (as judged by both cell-based and cell free LPL binding assays). Thus, the conserved cysteines in the Ly6 domain are crucial for GPIHBP1 function. PMID- 20026667 TI - Structure and epigenetics of nucleoli in comparison with non-nucleolar compartments. AB - The nucleolus is a nuclear compartment that plays an important role in ribosome biogenesis. Some structural features and epigenetic patterns are shared between nucleolar and non-nucleolar compartments. For example, the location of transcriptionally active mRNA on extended chromatin loop species is similar to that observed for transcriptionally active ribosomal DNA (rDNA) genes on so called Christmas tree branches. Similarly, nucleolus organizer region-bearing chromosomes located a distance from the nucleolus extend chromatin fibers into the nucleolar compartment. Specific epigenetic events, such as histone acetylation and methylation and DNA methylation, also regulate transcription of both rRNA- and mRNA-encoding loci. Here, we review the epigenetic mechanisms and structural features that regulate transcription of ribosomal and mRNA genes. We focus on similarities in epigenetic and structural regulation of chromatin in nucleoli and the surrounding non-nucleolar region and discuss the role of proteins, such as heterochromatin protein 1, fibrillarin, nucleolin, and upstream binding factor, in rRNA synthesis and processing. PMID- 20026668 TI - CD117-positive cells of the heart: progenitor cells or mast cells? AB - Human cardiac stem/progenitor cells and their potential for repair of heart injury are a current hot topic of research. CD117 has been used frequently as a marker for identification of stem/progenitor cells in the heart. However, cardiac mast cells, which are also CD117(+), have not been excluded by credible means when selecting putative cardiac progenitors by using CD117 as a marker. We evaluated the relationship between CD117(+) cells and mast cells in the left ventricle of human hearts (n=5 patients, ages 1 week-75 years) with the well established mast cell markers tryptase, toluidine blue, and thionine. A large number (85-100%) of CD117(+) cells in the human heart were specifically identified as mast cells. In addition, mast cells showed weak or moderate CD45 immunostaining signals. These results indicate that the majority of CD117(+) cells in the heart are mast cells and that these cells are distinctly positive for CD45, although staining was weak or moderate. These results strongly suggest that the newly reported CD117(+)/CD45(dim/moderate) putative cardiac progenitor cells are mast cells. The significance of this observation in stem cell research of the heart is discussed. PMID- 20026669 TI - Synthesis and organization of hyaluronan and versican by embryonic stem cells undergoing embryoid body differentiation. AB - Embryonic stem cells (ESCs) provide a convenient model to probe the molecular and cellular dynamics of developmental cell morphogenesis. ESC differentiation in vitro via embryoid bodies (EBs) recapitulates many aspects of early stages of development, including the epithelial-mesenchymal transition (EMT) of pluripotent cells into more differentiated progeny. Hyaluronan and versican are important extracellular mediators of EMT processes, yet the temporal expression and spatial distribution of these extracellular matrix (ECM) molecules during EB differentiation remains undefined. Thus, the objective of this study was to evaluate the synthesis and organization of hyaluronan and versican by using murine ESCs during EB differentiation. Hyaluronan and versican (V0 and V1 isoforms), visualized by immunohistochemistry and evaluated biochemically, accumulated within EBs during the course of differentiation. Interestingly, increasing amounts of a 70-kDa proteolytic fragment of versican were also detected over time, along with ADAMTS-1 and -5 protein expression. ESCs expressed each of the hyaluronan synthases (HAS) -1, -2, and -3 and versican splice variants (V0, V1, V2, and V3) throughout EB differentiation, but HAS-2, V0, and V1 were expressed at significantly increased levels at each time point examined. Hyaluronan and versican exhibited overlapping expression patterns within EBs in regions of low cell density, and versican expression was excluded from clusters of epithelial (cytokeratin-positive) cells but was enriched within the vicinity of mesenchymal (N-cadherin-positive) cells. These results indicate that hyaluronan and versican synthesized by ESCs within EB microenvironments are associated with EMT processes and furthermore suggest that endogenously produced ECM molecules play a role in ESC differentiation. This manuscript contains online supplemental material at http://www.jhc.org. Please visit this article online to view these materials. PMID- 20026670 TI - Retention of Pax3 expression in satellite cells of muscle spindles. AB - Intrafusal fibers within muscle spindles retain features characteristic of immaturity, unlike the larger and more numerous extrafusal fibers constituting the bulk of skeletal muscle. Satellite cells (SCs), myogenic progenitors, are detected on the surfaces of both intrafusal and extrafusal fibers, but little is known of spindle SCs. We have recently demonstrated that, like their extrafusal counterparts, SCs in muscle spindles of posthatch chickens express paired box transcription factor 7 (Pax7) protein. During vertebrate embryogenesis, myogenic progenitors express both Pax7 and Pax3 proteins. In postnatal mice, Pax3 appears in rare SC subsets, whereas Pax7 is expressed by all SCs within extrafusal fibers. Here we test the hypothesis that Pax3 protein maintains localized expression within SCs of muscle spindles. Immunohistochemical techniques were used to identify SCs by their Pax7 expression within anterior latissimus dorsi muscle excised from posthatch chickens of various ages. A greater percentage of SCs express Pax3 within intrafusal than extrafusal fibers at each age, and the proportion of SCs expressing Pax3 declines with aging. This is the first study to localize Pax3 expression in posthatch avian muscle and within SCs of muscle spindles. We suggest that Pax3-positive SCs are involved in fiber maintenance. PMID- 20026671 TI - Immunocytochemical localization of sex steroid hormone receptors in normal human mammary gland. AB - The sex steroids, estrogens, progesterone, and androgens, all play a role in mammary development and function. To precisely identify the sites of action of these steroids, we studied the localization of the estrogen receptor alpha (ERalpha) and ERbeta, the progesterone receptor A (PRA) and PRB, and androgen receptors (AR) in the normal human mammary gland. Immunocytochemical localization of ERalpha, ERbeta, PRA, PRB, and AR was performed with reduction mammoplasty specimens from premenopausal women. ERalpha, PRA, PRB, and AR were localized mostly to the inner layer of epithelial cells lining acini and intralobular ducts, as well as to myoepithelial cells scattered in the external layer of interlobular ducts. AR was also found in some stromal cells. ERbeta staining was more widespread, resulting in epithelial and myoepithelial cells being labeled in acini and ducts as well as stromal cells. These results suggest that all sex steroids can directly act on epithelial cells to modulate development and function of the human mammary gland. Estrogens and androgens can also indirectly influence epithelial cell activity by an action on stromal cells. PMID- 20026672 TI - Fixative composition alters distributions of immunoreactivity for glutaminase and two markers of nociceptive neurons, Nav1.8 and TRPV1, in the rat dorsal root ganglion. AB - Most, if not all, dorsal root ganglion (DRG) neurons use the neurotransmitter glutamate. There are, however, conflicting reports of the percentages of DRG neurons that express glutaminase (GLS), the enzyme that synthesizes glutamate, ranging from 30% to 100% of DRG neurons. Defining DRG neuron populations by the expression of proteins like GLS, which indicates function, is routinely accomplished with immunolabeling techniques. Proper characterization of DRG neuron populations relies on accurate detection of such antigens. It is known intuitively that fixation can alter immunoreactivity (IR). In this study, we compared the effects of five formaldehyde concentrations between 0.25% and 4.0% (w/v) and five picric acid concentrations between 0.0% and 0.8% (w/v) on the IR of GLS, the voltage-gated sodium channel 1.8 (Na(v)1.8), and the capsaicin receptor TRPV1. We also compared the effects of five incubation time lengths from 2 to 192 hr, in primary antiserum on IR. Lowering formaldehyde concentration elevated IR for all three antigens, while raising picric acid concentration increased Na(v)1.8 and TRPV1 IR. Increasing IR improved detection sensitivity, which led to higher percentages of labeled DRG neurons. By selecting fixation conditions that optimized IR, we found that all DRG neurons express GLS, 69% of neurons express Na(v)1.8, and 77% of neurons express TRPV1, indicating that some previous studies may have underestimated the percentages of DRG neurons expressing these proteins. This manuscript contains online supplemental material at http://www.jhc.org. Please visit this article online to view these materials. PMID- 20026673 TI - Antigen-specific suppression of experimental autoimmune encephalomyelitis by a novel bifunctional peptide inhibitor: structure optimization and pharmacokinetics. AB - The objective of this study was to optimize the in vivo activity of proteolipid protein (PLP)-bifunctional peptide inhibitor (BPI) molecule to suppress experimental autoimmune encephalomyelitis (EAE) in SJL/J mice and evaluate pharmacokinetic profiles of PLP-BPI. PLP-BPI is constructed via conjugation of myelin PLP(139-151) with CD11a(237-246)-derived peptide (LABL) via a spacer. The hypothesis is that PLP-BPI binds simultaneously to major histocompatibility complex-II and intercellular adhesion molecule-1 on the antigen-presenting cell (APC) and inhibits the formation of the immunological synapse during T-cell and APC interactions. In this study, the structure of BPI was modified by varying the spacer and was evaluated in the EAE model. Intravenous injections of BPI derivatives inhibited the onset, severity, and incidence of EAE more effectively and induced a lower incidence of anaphylaxis than that produced by unmodified PLP BPI. As anticipated, production of interleukin-17, a proinflammatory cytokine commonly found in elevated levels among multiple sclerosis (MS) patients, was significantly lower in Ac-PLP-BPI-PEG6- or Ac-PLP-BPI-NH(2)-2-treated mice than in phosphate-buffered saline-treated mice. These results suggest that BPI-type molecules can be modified to achieve more efficient and better tolerated BPI based derivatives for the treatment of MS. PMID- 20026675 TI - A hybrid indoloquinolizidine peptide as allosteric modulator of dopamine D1 receptors. AB - The indoloquinolizidine-peptide 28 [(3S,12bR)-N-((S)-1-((S)-1-((S)-2 carbamoylpyrrolidin-1-yl)-3-(4-fluorophenyl)-1-oxopropan-2-ylamino)-4-cyclohexyl 1-oxobutan-2-yl)-1,2,3,4,6,7,12, 12b-octahydroindolo[2,3-a]quinolizine-3 carboxamide], a trans-indoloquinolizidine-peptide hybrid obtained by a combinatorial approach, behaved as an orthosteric ligand of all dopamine D(2) like receptors (D(2), D(3), and D(4)) and dopamine D(5) receptors, but as a negative allosteric modulator of agonist and antagonist binding to striatal dopamine D(1) receptors. Indoloquinolizidine-peptide 28 induced a concentration dependent hyperbolic increase in the antagonist apparent equilibrium dissociation constant values and altered the dissociation kinetics of dopamine D(1) receptor antagonists. The negative allosteric modulation was also found when agonist binding to D(1) receptors was assayed. Indoloquinolizidine-peptide 28 was a weak ago-allosteric modulator but markedly led to a decreased potency without decreasing the maximum partial/full agonist-mediated effect on cAMP levels. Compounds able to decrease the potency while preserving the efficacy of D(1) receptor agonists are promising for exploration in psychotic pathologies. PMID- 20026674 TI - Antillatoxin, a novel lipopeptide, enhances neurite outgrowth in immature cerebrocortical neurons through activation of voltage-gated sodium channels. AB - Antillatoxin (ATX) is a structurally novel lipopeptide that activates voltage gated sodium channels (VGSC) leading to sodium influx in cerebellar granule neurons and cerebrocortical neurons 8 to 9 days in vitro (Li et al., 2001; Cao et al., 2008). However, the precise recognition site for ATX on the VGSC remains to be defined. Inasmuch as elevation of intracellular sodium ([Na(+)](i)) may increase N-methyl-d-aspartate receptor (NMDAR)-mediated Ca(2+) influx, Na(+) may function as a signaling molecule. We hypothesized that ATX may enhance neurite outgrowth in cerebrocortical neurons by elevating [Na(+)](i) and augmenting NMDAR function. ATX (30-100 nM) robustly stimulated neurite outgrowth, and this enhancement was sensitive to the VGSC antagonist, tetrodotoxin. To unambiguously demonstrate the enhancement of NMDA receptor function by ATX, we recorded single channel currents from cell-attached patches. ATX was found to increase the open probability of NMDA receptors. Na(+)-dependent up-regulation of NMDAR function has been shown to be regulated by Src family kinase (SFK) (Yu and Salter, 1998). The Src kinase inhibitor PP2 abrogated ATX-enhanced neurite outgrowth, suggesting a SFK involvement in this response. ATX-enhanced neurite outgrowth was also inhibited by the NMDAR antagonist, (5R,10S)-(+)-5-methyl-10,11-dihydro-5H dibenzo[a,d]cyclohepten-5,10-imine hydrogen maleate (MK-801), and the calmodulin dependent kinase kinase (CaMKK) inhibitor, 1,8-naphthoylene benzimidazole-3 carboxylic acid (STO-609), demonstrating the requirement for NMDAR activation with subsequent downstream engagement of the Ca(2+)-dependent CaMKK pathway. These results with the structurally and mechanistically novel natural product, ATX, confirm and generalize our earlier results with a neurotoxin site 5 ligand. These data suggest that VGSC activators may represent a novel pharmacological strategy to regulate neuronal plasticity through NMDAR-dependent mechanisms. PMID- 20026678 TI - On the classification of epistatic interactions. AB - Modern genomewide association studies are characterized by the problem of "missing heritability." Epistasis, or genetic interaction, has been suggested as a possible explanation for the relatively small contribution of single significant associations to the fraction of variance explained. Of particular concern to investigators of genetic interactions is how to best represent and define epistasis. Previous studies have found that the use of different quantitative definitions for genetic interaction can lead to different conclusions when constructing genetic interaction networks and when addressing evolutionary questions. We suggest that instead, multiple representations of epistasis, or epistatic "subtypes," may be valid within a given system. Selecting among these epistatic subtypes may provide additional insight into the biological and functional relationships among pairs of genes. In this study, we propose maximum-likelihood and model selection methods in a hypothesis-testing framework to choose epistatic subtypes that best represent functional relationships for pairs of genes on the basis of fitness data from both single and double mutants in haploid systems. We gauge the performance of our method with extensive simulations under various interaction scenarios. Our approach performs reasonably well in detecting the most likely epistatic subtype for pairs of genes, as well as in reducing bias when estimating the epistatic parameter (epsilon). We apply our approach to two available data sets from yeast (Saccharomyces cerevisiae) and demonstrate through overlap of our identified epistatic pairs with experimentally verified interactions and functional links that our results are likely of biological significance in understanding interaction mechanisms. We anticipate that our method will improve detection of epistatic interactions and will help to unravel the mysteries of complex biological systems. PMID- 20026677 TI - Mutational analysis of Candida albicans SNF7 reveals genetically separable Rim101 and ESCRT functions and demonstrates divergence in bro1-domain protein interactions. AB - The opportunistic pathogen Candida albicans can grow over a wide pH range, which is associated with its ability to colonize and infect distinct host niches. C. albicans growth in neutral-alkaline environments requires proteolytic activation of the transcription factor Rim101. Rim101 activation requires Snf7, a member of the endosomal sorting complex required for transport (ESCRT) pathway. We hypothesized that Snf7 has distinct functions in the Rim101 and ESCRT pathways, which we tested by alanine-scanning mutagenesis. While some snf7 alleles conferred no defects, we identified alleles with solely ESCRT-dependent, solely Rim101-dependent, or both Rim101- and ESCRT-dependent defects. Thus, Snf7 function in these two pathways is at least partially separable. Both Rim101- and ESCRT-dependent functions require Snf7 recruitment to the endosomal membrane and alleles that disrupted both pathways were found to localize normally, suggesting a downstream defect. Most alleles that conferred solely Rim101-dependent defects were still able to process Rim101 normally under steady-state conditions. However, these same strains did display a kinetic defect in Rim101 processing. Several alleles with solely Rim101-dependent defects mapped to the C-terminal end of Snf7. Further analyses suggested that these mutations disrupted interactions with bro-domain proteins, Rim20 and Bro1, in overlapping but slightly divergent Snf7 domains. PMID- 20026679 TI - Regulation by blue light of the fluffy gene encoding a major regulator of conidiation in Neurospora crassa. AB - The development of asexual spores, that is, the process of conidiation, in the fungus Neurospora crassa is increased by light. The fluffy (fl) gene, encoding a major regulator of conidiation, is activated by light. We describe here a detailed characterization of the regulation by blue light of fl in vegetative hyphae. This induction requires the white collar complex (WCC) while the FLD protein acts as a dark repressor of fl transcription. We show that the WCC directly regulates fl transcription in response to blue light after transiently binding the promoter. We propose that fl is repressed by FLD in vegetative mycelia and that the repression is lost after light exposure and WCC activation. The increase in fl mRNA in vegetative mycelia after light exposure, and the corresponding increase in the amount of the regulatory FL protein, should promote the activation of the conidiation pathway. The activation by light of fl provides a simple mechanism for the activation of conidiation by blue light in Neurospora that may be at work in other fungi. PMID- 20026680 TI - Gag proteins of Drosophila telomeric retrotransposons: collaborative targeting to chromosome ends. AB - TAHRE, the least abundant of the three retrotransposons forming telomeres in Drosophila melanogaster, has high sequence similarity to the gag gene and untranslated regions of HeT-A, the most abundant telomere-specific retrotransposon. Despite TAHRE's apparent evolutionary relationship to HeT-A, we find TAHRE Gag cannot locate to telomere-associated "Het dots" unless collaborating with HeT-A Gag. TAHRE Gag is carried into nuclei by HeT-A or TART Gag, but both TART and TAHRE Gags need HeT-A Gag to localize to Het dots. When coexpressed with the appropriate fragment of HeT-A and/or TART Gags, TAHRE Gag multimerizes with either protein. HeT-A and TART Gags form homo- and heteromultimers using a region containing major homology region (MHR) and zinc knuckle (CCHC) motifs, separated by a pre_C2HC motif (motifs common to other retroelements). This region's sequence is strongly conserved among the three telomeric Gags, with precise spacing of conserved residues. Nontelomeric Gags neither interact with the telomeric Gags nor have this conserved spacing. TAHRE Gag is much less able to enter the nucleus by itself than HeT-A or TART Gags. The overall telomeric localization efficiency for each of the three telomeric Gag proteins correlates with the relative abundance of that element in telomere arrays, suggesting an explanation for the relative rarity of TAHRE elements in telomere arrays and supporting the hypothesis that Gag targeting to telomeres is important for the telomere-specific transposition of these elements. PMID- 20026681 TI - A neo-sex chromosome that drives postzygotic sex determination in the hessian fly (Mayetiola destructor). AB - Two nonoverlapping autosomal inversions defined unusual neo-sex chromosomes in the Hessian fly (Mayetiola destructor). Like other neo-sex chromosomes, these were normally heterozygous, present only in one sex, and suppressed recombination around a sex-determining master switch. Their unusual properties originated from the anomalous Hessian fly sex determination system in which postzygotic chromosome elimination is used to establish the sex-determining karyotypes. This system permitted the evolution of a master switch (Chromosome maintenance, Cm) that acts maternally. All of the offspring of females that carry Cm-associated neo-sex chromosomes attain a female-determining somatic karyotype and develop as females. Thus, the chromosomes act as maternal effect neo-W's, or W-prime (W') chromosomes, where ZW' females mate with ZZ males to engender female-producing (ZW') and male-producing (ZZ) females in equal numbers. Genetic mapping and physical mapping identified the inversions. Their distribution was determined in nine populations. Experimental matings established the association of the inversions with Cm and measured their recombination suppression. The inversions are the functional equivalent of the sciarid X-prime chromosomes. We speculate that W' chromosomes exist in a variety of species that produce unisexual broods. PMID- 20026676 TI - Natural variation in odorant recognition among odorant-binding proteins in Drosophila melanogaster. AB - Chemical recognition is essential for survival and reproduction. Adaptive evolution has resulted in diverse chemoreceptor families, in which polymorphisms contribute to individual variation in chemosensation. To gain insights into the genetic determinants of individual variation in odorant recognition, we measured olfactory responses to two structurally similar odorants in a population of wild derived inbred lines of Drosophila melanogaster. Odorant-binding proteins (OBPs) are the first components of the insect olfactory system to encounter odorants. Previously four single-nucleotide polymorphisms (SNPs) in the Obp99 group were associated with variation in olfactory responses to benzaldehyde. Here, we identify six different SNPs that are associated with variation in responses to a structurally similar odorant, acetophenone, in the same Obp genes. Five SNPs are in coding regions of Obp99b and Obp99d and one SNP is in the 3'-untranslated region of Obp99a (A610G). We found that the 610G allele is associated with higher response scores to acetophenone than the 610A allele, but with lower expression of Obp99a, suggesting that binding of acetophenone to Opb99a might limit rather than facilitate access to odorant receptors. Our results show that overlapping sets of OBPs contribute to odorant recognition for structurally similar odorants, but that different SNPs are associated with odorant-specific individual variation. Thus, dual olfactory recognition where OBPs regulate odorant access to receptors may enhance olfactory discrimination. PMID- 20026683 TI - Identification of novel hair-growth inducers by means of connectivity mapping. AB - The aim of this study was to identify novel inducers of hair growth using gene expression profiling at various stages of hair-growth induction. First, we analyzed gene expression at the onset of hair growth in mice induced by cyclosporin A (CsA), a well-known hair-growth inducer, using DNA microarray analysis. The results unveiled genes involved in the step-by-step progression of hair growth, including increases in melanin biosynthesis and decreases in immune response at d 2 and the subsequent stimulation of cell proliferation at d 4, followed by the up-regulation of hair specific keratins at d 7 after CsA treatment. With the use of the connectivity map (Cmap), agents that had a similar "gene signature" to that of the profiles of CsA-treated mice were identified. Several agents, including CsA, were identified by the Cmap and were evaluated for hair induction activity in vivo. One of the proposed agents, fluphenazine (from the d 2 signature) actually induced hair growth in vivo (ED(50): 2 mM for single application), and the subsequent application of 5 mM iloprost (from the d 4 signature) significantly enhanced the hair-growth effect of fluphenazine. From these results, Cmap analysis was proven to be a useful method that connects gene expression profiles of complicated biological processes, such as hair-growth induction, to effective agents. PMID- 20026682 TI - Silencing of the JNK pathway maintains progesterone receptor activity in decidualizing human endometrial stromal cells exposed to oxidative stress signals. AB - Survival of the conceptus is dependent on continuous progesterone signaling in the maternal decidua but how this is achieved under conditions of oxidative stress that characterize early pregnancy is unknown. Using primary cultures, we show that modest levels of reactive oxygen species (ROS) increase sumoylation in human endometrial stromal cells (HESCs), leading to enhanced modification and transcriptional inhibition of the progesterone receptor (PR). The ability of ROS to induce a sustained hypersumoylation response, or interfere with PR activity, was lost upon differentiation of HESCs into decidual cells. Hypersumoylation in response to modest levels of ROS requires activation of the JNK pathway. Although ROS-dependent JNK signaling is disabled on decidualization, the cells continue to mount a transcriptional response, albeit distinct from that observed in undifferentiated HESCs. We further show that attenuated JNK signaling in decidual cells is a direct consequence of altered expression of key pathway modulators, including induction of MAP kinase phosphatase 1 (MKP1). Overexpression of MKP1 dampens JNK signaling, prevents hypersumoylation, and maintains PR activity in undifferentiated HESCs exposed to ROS. Thus, JNK silencing uncouples ROS signaling from the SUMO conjugation pathway and maintains progesterone responses and cellular homeostasis in decidual cells under oxidative stress conditions imposed by pregnancy. PMID- 20026685 TI - Ascribing intentions in clinical decision-making. AB - BACKGROUND: The intentions of clinicians are widely considered to be relevant to the ethical assessment of their actions. A better understanding of the psychological factors that influence the ascription of intentions in clinical practice is important for improving the self-understanding of clinical decision making and, ultimately, the ethics of clinical care. Drawing on empirical research on intentionality that has been done in other contexts, this is the first study to test whether the "asymmetric effect" of intention ascription is exhibited by respondents when presented with clinical decision-making scenarios. OBJECTIVE: To assess how individuals attribute intentions to clinical actors in clinical decision-making scenarios. METHODS: A total of 149 first and second year medical students was randomly assigned to two groups (group A, group B). Subjects in each group read two scenarios and submitted anonymous responses to questions regarding each scenario. RESULTS: The asymmetric effect was strongly exhibited by the responses given to scenario 2, but it was not exhibited by the responses given to scenario 1. CONCLUSION: The present study provided evidence for the view that people's ascription of intentions to others is influenced by their previous evaluative judgement of the conduct in question. PMID- 20026686 TI - "It's crucial they're treated as patients": ethical guidance and empirical evidence regarding treating doctor-patients. AB - Ethical guidance from the British Medical Association (BMA) about treating doctor patients is compared and contrasted with evidence from a qualitative study of general practitioners (GPs) who have been patients. Semistructured interviews were conducted with 17 GPs who had experienced a significant illness. Their experiences were discussed and issues about both being and treating doctor patients were revealed. Interpretative phenomenological analysis was used to evaluate the data. In this article data extracts are used to illustrate and discuss three key points that summarise the BMA ethical guidance, in order to develop a picture of how far experiences map onto guidance. The data illustrate and extend the complexities of the issues outlined by the BMA document. In particular, differences between experienced GPs and those who have recently completed their training are identified. This analysis will be useful for medical professionals both when they themselves are unwell and when they treat doctor patients. It will also inform recommendations for professionals who educate medical students or trainees. PMID- 20026687 TI - Healthcare workers' perceptions of the duty to work during an influenza pandemic. AB - Healthcare workers (HCWs) are often assumed to have a duty to work, even if faced with personal risk. This is particularly so for professionals (doctors and nurses). However, the health service also depends on non-professionals, such as porters, cooks and cleaners. The duty to work is currently under scrutiny because of the ongoing challenge of responding to pandemic influenza, where an effective response depends on most uninfected HCWs continuing to work, despite personal risk. This paper reports findings of a survey of HCWs (n = 1032) conducted across three National Health Service trusts in the West Midlands, UK, to establish whether HCWs' likelihood of working during a pandemic is associated with views about the duty to work. The sense that HCWs felt that they had a duty to work despite personal risk emerged strongly regardless of professional status. Besides a strong sense that everyone should pull together, all kinds of HCWs recognised a duty to work even in difficult circumstances, which correlated strongly with their stated likelihood of working. This suggests that HCWs' decisions about whether or not they are prepared to work during a pandemic are closely linked to their sense of duty. However, respondents' sense of the duty to work may conflict with their sense of duty to family, as well as other factors such as a perceived lack of reciprocity from their employers. Interestingly, nearly 25% of doctors did not consider that they had a duty to work where doing so would pose risks to themselves or their families. PMID- 20026688 TI - When physicians forego the doctor-patient relationship, should they elect to self prescribe or curbside? An empirical and ethical analysis. AB - BACKGROUND: The American Medical Association, the British Medical Association and the Canadian Medical Association have guidelines that specifically discourage physicians from self-prescribing or prescribing to family members, but only the BMA addresses informal prescription requests between colleagues. OBJECTIVE: To examine the practices of paediatric providers regarding self-prescribing, curbsiding colleagues, and prescribing and refusing to prescribe to friends and family. METHODS: 1086 paediatricians listed from the American Academy of Paediatrics 2007 web-based directory were surveyed. RESULTS: 44% (430/982) of eligible survey respondents returned usable surveys. Almost half (198/407) of respondents had prescribed for themselves. An equal number (198/411) had informally requested a prescription from a colleague. Three-quarters (325/429) stated they had been asked to prescribe a prescription drug for a first-degree or second-degree relative, and 51% (186/363) had been asked by their spouse. Eighty six per cent (343/397) stated that they had refused to write a prescription on at least one occasion for a friend or family member. The following reasons "strongly influenced" their decision to refuse a prescription request: (1) outside of provider's expertise (88%); (2) patient's need for his or her own physician (70%); (3) not medically indicated (69%); (4) need for a physical examination (65%). CONCLUSION: These data confirm that most physicians have engaged in self prescribing or curbside requests for prescriptions. It can be argued that curbsiding is more morally problematic than self-prescribing because it implicates a third party, and should be discouraged regardless of whether the requester is a colleague, family member or friend. PMID- 20026689 TI - Physicians' labelling of end-of-life practices: a hypothetical case study. AB - OBJECTIVES: To investigate why physicians label end-of-life acts as either 'euthanasia/ending of life' or 'alleviation of symptoms/palliative or terminal sedation', and to study the association of such labelling with intended reporting of these acts. METHODS: Questionnaires were sent to a random, stratified sample of 2100 Dutch physicians (response: 55%). They were asked to label six hypothetical end-of-life cases: three 'standard' cases and three cases randomly selected (out of 47), that varied according to (1) type of medication, (2) physician's intention, (3) type of patient request, (4) patient's life expectancy and (5) time until death. We identified the extent to which characteristics of cases are associated with physician's labelling, with multilevel multivariable logistic regression. RESULTS: The characteristics that contributed most to labelling cases as 'euthanasia/ending of life' were the administration of muscle relaxants (99% of these cases were labelled as 'euthanasia/ending of life') or disproportional morphine (63% of these cases were labelled accordingly). Other important factors were an intention to hasten death (54%) and a life expectancy of several months (46%). Physicians were much more willing to report cases labelled as 'euthanasia' (87%) or 'ending of life' (56%) than other cases. CONCLUSIONS: Similar cases are not uniformly labelled. However, a physicians' label is strongly associated with their willingness to report their acts. Differences in how physicians label similar acts impede complete societal control. Further education and debate could enhance the level of agreement about what is physician-assisted dying, and thus should be reported, and what not. PMID- 20026690 TI - The right not to know and preimplantation genetic diagnosis for Huntington's disease. AB - The right not to know is underappreciated in policy-making. Despite its articulation in medical law and ethics, policy-makers too easily let other concerns override the right not to know. This observation is triggered by a recent decision of the Dutch government on embryo selection for Huntington's disease. This is a monogenetic debilitating disease without cure, leading to death in early middle age, and thus is a likely candidate for preimplantation genetic diagnosis (PGD). People possibly affected with the Huntington gene do not always want to know their genetic status themselves, which is very burdensome knowledge. For prospective parents, this creates a difficulty in having unaffected children through embryo selection without knowing their own genetic status. A solution is provided by the exclusion test: it allows embryo selection while honouring the parents' right not to know. The Dutch government however disallowed the exclusion test and chose to allow PGD on Huntington only for parents who test themselves first. To avoid "unnecessary" in-vitro fertilisation procedures for unaffected parents, prospective parents are "forced to know" before they can conceive through embryo selection. This article analyses the scope of the right not to know in the context of embryo selection against Huntington's disease. It concludes that the right not to know implies that PGD against Huntington should be allowed by means of the exclusion test. PMID- 20026691 TI - Payment for participation in research: a pursuit for the poor? AB - Poor people predominate as a subgroup of those who take part in healthy volunteer research. They are subjected to minimised but unknown risks and unpleasant burdens so that the safety of new medicines can be evaluated. This is prima facie unfair especially given that the poor are often unable to access expensive medicines. Although participants in this kind of research often do receive compensation for their time, these payments are usually capped at a very low level. This paper defends a version of a reimbursement model for the payment of research subjects. This model is practical, would benefit those without an income who take part in research, and would make it possible for those in regular work to take part in phase 1 research. PMID- 20026692 TI - An investigation of patients' motivations for their participation in genetics related research. AB - DESIGN: Qualitative interview study. PARTICIPANTS: Fifty-nine patients with a family history of cancer who attend a regional cancer genetics clinic in the UK were interviewed about their current and previous research experiences. FINDINGS: Interviewees gave a range of explanations for research participation. These were categorised as (a) social--research participation benefits the wider society by progressing science and improving treatment for everyone; (b) familial--research participation may improve healthcare and benefit current or future generations of the participant's family; and (c) personal--research participation provides therapeutic or non-therapeutic benefits for oneself. CONCLUSIONS: We discuss the distinction drawn between motives for research participation focused upon self (personal) and others (familial/social), and observe that personal, social and familial motives can be seen as interdependent. For example, research participation that is undertaken to benefit others, particularly relatives, may also offer a number of personal benefits for self, such as enabling participants to feel that they have discharged their social or familial obligations. We argue for the need to move away from simple, static, individualised notions of research participation to a more complex, dynamic and inherently social account. PMID- 20026693 TI - The role of 'public opinion' in the UK animal research debate. AB - Animal research remains a deeply controversial topic in biomedical science. While a vast amount has been written about the ethical status of laboratory animals, far less academic attention has been devoted to the public and, more specifically, to public opinion. Rather than what the public think, this article considers the role of 'public opinion'. It draws on a recent empirical study which involved interviews with laboratory scientists who use animals in their research, and with other UK stakeholders. The first section of the paper demonstrates that public opinion has become a kind of resource in the animal research debate. Public opinion polls, in particular, are frequently cited. The second section explores this further and argues that, for all sides, appealing to public opinion is a key way to show legitimacy. Finally, the paper shifts gear to consider whether public opinion should matter, both for ethical reasoning and for science policy. PMID- 20026694 TI - A practical approach to teaching medical ethics. AB - Teaching medical ethics and law has become much more prominent in medical student education, largely as a result of a 1998 consensus statement on such teaching. Ethics is commonly taught at undergraduate level using lectures and small group tutorials, but there is no recognised method for transferring this theoretical knowledge into practice and ward-based learning. This reflective article by a Sheffield university undergraduate medical student describes the value of using a student-selected component to study practical clinical ethics and the use of a clinical ethics checklist. The ethical checklist was proposed by Sokol as a tool for use by medical staff during the ward round to prompt the consideration of important ethical principles in relation to care. This paper describes additional uses for the checklist as a tool for teaching and learning about the practical application of ethical principles and for observing professional behaviours within a critical care and acute care environment. Evidence suggests that putting ethical behaviour into practice offers a far greater challenge to a newly qualified doctor than has been appreciated, and that more needs to be done at an undergraduate level to help combat this. This paper argues from a personal standpoint of an individual student experience that this can best be done in a clinical medical setting. PMID- 20026695 TI - Medical ethics and law for doctors of tomorrow: the 1998 Consensus Statement updated. AB - Knowledge of the ethical and legal basis of medicine is as essential to clinical practice as an understanding of basic medical sciences. In the UK, the General Medical Council (GMC) requires that medical graduates behave according to ethical and legal principles and must know about and comply with the GMC's ethical guidance and standards. We suggest that these standards can only be achieved when the teaching and learning of medical ethics, law and professionalism are fundamental to, and thoroughly integrated both vertically and horizontally throughout, the curricula of all medical schools as a shared obligation of all teachers. The GMC also requires that each medical school provides adequate teaching time and resources to achieve the above. We reiterate that the adequate provision and coordination of teaching and learning of ethics and law requires at least one full-time senior academic in ethics and law with relevant professional and academic expertise. In this paper we set out an updated indicative core content of learning for medical ethics and law in UK medical schools and describe its origins and the consultative process by which it was achieved. PMID- 20026696 TI - Rights, respect for dignity and end-of-life care: time for a change in the concept of informed consent. AB - The current concepts of autonomy, surrogate autonomy and informed consent often lead to futile and expensive care at the ends of life. They may impinge on the dignity of the patient as well as subject society to unwarranted expense. In order to provide affordable healthcare for all, these concepts are in need of modification. PMID- 20026697 TI - Are kids having a rough time of it in sports? PMID- 20026698 TI - Dieting to win or to be thin? A study of dieting and disordered eating among adolescent elite athletes and non-athlete controls. AB - OBJECTIVE: To examine the prevalence of dieting, reasons for dieting and prevalence of disordered eating among adolescent elite athletes and age-matched controls, and to examine the differences between athletes competing in leanness and non-leanness sports. METHODS: First-year students of 16 different Norwegian Elite Sport High Schools (athlete group, n = 682) and two randomly selected ordinary high schools from a county representative of the general Norwegian population (control group, n = 423) were invited to participate in this cross sectional study. A total of 606 athletes and 355 controls completed the questionnaire, giving a response rate of 89% and 84%, respectively. The questionnaire contained questions regarding training patterns, menstrual status and history, dieting, use of pathogenic weight control methods and the drive for thinness (DT) and body dissatisfaction (BD) subscales from the Eating Disorders Inventory. MAIN OUTCOME MEASURE: Disordered eating, defined as meeting one or more of the following criteria: DT score > or =15 (girls) and > or =10 (boys), BD score > or =14 (girls) and > or =10 (boys), body mass index <17.9 kg/m(2) (girls) and <17.5 kg/m(2) (boys), current and/or > or =3 previous efforts to lose weight, use of pathogenic weight control methods and self-reported menstrual dysfunction. RESULTS: A higher prevalence of control subjects were dieting and classified with disordered eating compared with the athletes. An improvement of appearance was a more common reason for dieting among controls compared with athletes. No differences in dieting or disordered eating were found between leanness and non leanness sports athletes. CONCLUSIONS: Self-reported disordered eating is more prevalent among controls than adolescent elite athletes, and losing weight to enhance performance is an important reason for dieting among adolescent elite athletes. PMID- 20026699 TI - BJSM reviews: A-Z of nutritional supplements: dietary supplements, sports nutrition foods and ergogenic aids for health and performance Part 5. PMID- 20026700 TI - Similar immunogenetics of Barrett's oesophagus and cervical neoplasia: is HPV the common denominator? PMID- 20026701 TI - HLA and kidney transplantation. AB - This article focuses on the immunogenetics, immunology, rejection and immunosuppression in kidney transplantation. HLA matching still affects outcome data, and HLA matching improves graft survival. Graft sources and related outcomes are discussed. PMID- 20026702 TI - Immunophenotypic distinction between pigmented villonodular synovitis and haemosiderotic synovitis. AB - AIM: Haemosiderotic synovitis (HS) is caused by excessive bleeding into a joint. It occurs secondary to a variety of conditions and needs to be distinguished from pigmented villonodular synovitis (PVNS) for the purposes of treatment. The histopathological distinction between these conditions, particularly in biopsy specimens, can be problematic. METHODS: Immunophenotypic findings in 20 cases of PVNS and 20 cases of HS were analysed using monoclonal antibodies against proliferation (Ki-67), apoptosis (bcl2), macrophage (CD14, CD68, HLA-DR) and osteoclast (CD51) antigens. RESULTS: Macrophages in PVNS and HS expressed CD14 and HLA-DR. The giant cells in PVNS, but not those in HS, expressed CD51 and were negative for CD14 and HLA-DR, indicating that these cells had an osteoclast phenotype. Considerably more CD51-expressing mononuclear cells were noted in PVNS compared with HS. The Ki-67 proliferation index was higher in PVNS than in HS. CONCLUSIONS: The findings indicate that there are immunophenotypic differences in giant cells between PVNS and HS, and that expression of CD51 and a high Ki-67 index effectively distinguishes between these two conditions. PMID- 20026703 TI - Correlation of autoantibodies and CD5+ B cells in ocular adnexal marginal zone B cell lymphomas. AB - AIM: To determine the clinicopathological properties of ocular adnexal marginal zone B cell lymphomas (MZBLs) with CD5+ B cells. METHODS: This study determined the clinicopathological properties of MZBL samples from 97 patients with ocular adnexal MZBLs and searched for hallmarks of systemic autoimmunity in these patients. RESULTS: Two elderly female patients were found to have ocular adnexal MZBLs with CD5+ B cells; flow cytometry analysis suggested that one of these MZBLs had CD5+ B cell clonal proliferation. The levels of anti-single stranded (SS)-DNA and anti-SS-A/Ro antibodies in these two patients were significantly higher than those in controls that were matched for age, gender and disease (2/2 versus 0/14; p = 0.008) and controls without MZBL (2/2 versus 0/30; p = 0.002). The genes from the immunoglobulin heavy-chain variable region for one of the patients showed a V3-21 segment. In addition, another patient with ocular adnexal reactive lymphoid hyperplasia with CD5+ B cells also had anti-SS-DNA antibodies. CONCLUSION: Patients with ocular adnexal MZBLs with CD5+ B cells may have a background of systemic conditions with CD5+ B-cell-related autoantibodies. PMID- 20026704 TI - Female-type fibrocystic disease with papillary hyperplasia in a male breast. AB - Fibrocystic disease is a common benign finding in the female breast and often presents as a palpable mass. It is much less commonly found in the male breast. A case is reported of a young man with female-type fibrocystic disease associated with papillary hyperplasia in the right breast. PMID- 20026705 TI - Retraction. "Tracking the footprints of the rabies virus: are we any closer to decoding this elusive virus?" by Mahadevan A, Suja MS, Madhusudana SN, et al. PMID- 20026707 TI - Ossifying fibroma: a systematic review. AB - OBJECTIVES: The aim was to evaluate the principal features of ossifying fibroma (OF) by systematic review (SR), and to compare their frequencies among four global groups. METHODS: The databases searched were the PubMed interface of MEDLINE and LILACS. Only those reports of OFs that occurred in a series in the reporting authors' caseload were considered. All cases were confirmed fibro osseous lesions histopathologically. The SR-included series had also to have used radiographs. RESULTS: Of the 64 reports (including the Hong Kong report) considered, 32 reports and a total of 781 cases were included in the SR. Ten SR included series were in languages other than English. OF affected females more frequently, but was three times more prevalent in the mandible. The mean age at first presentation was 31 years. The decade with the greatest frequency was the fourth. Females were in the majority except in the first decade. The main symptom in 66% of all SR-included cases was swelling (including deformation of the jaws). 31% were found incidentally. 84% of cases displayed buccolingual expansion; half of the mandibular cases exhibited downward displacement of the lower border of the mandible and 90% of maxillary cases involved the maxillary antrum. Only 28% of reports included follow-up; 12% of cases recurred or were reactivated. CONCLUSIONS: Although long-term follow-up of large series that would have revealed the long-term outcomes of OF was lacking, a 12% recurrence rate is clinically significant and suggests that OFs should be considered for long-term follow-up. PMID- 20026708 TI - Ossifying fibroma in a Hong Kong community: the clinical and radiological features and outcomes of treatment. AB - OBJECTIVES: The aim of this study was to evaluate the principal clinical and radiological features of a consecutive series of cases of ossifying fibroma (OF) affecting a Hong Kong Chinese community and to determine their outcome by follow up. METHODS: All cases were accompanied by appropriate radiography and were histopathologically confirmed as fibro-osseous lesions. RESULTS: The clinical and radiological presentations, differential diagnoses and outcomes of follow-up of 24 consecutive patients with OFs were reviewed. All patients were female. The mean age at first presentation was 38.0 years. The oldest reported patient first presenting with OF was 80 years old. Most OFs (58%) were discovered incidentally. The maxilla and mandible were affected in 4 and 20 cases, respectively. Only 7 cases, out of the 15 cases for which provisional diagnosis was available, included OF. 16 patients were followed up for a mean of 5.3 years, of whom 12 discharged themselves after a mean of 3.0 years. Only one lesion recurred. CONCLUSIONS: OF in this community predominantly affects females. Although gynaecological history was not included in the clinical notes, there is a clear possibility of reactivation of OF growth at the menopause. Other potentially important contributing factors are "traditional Chinese medicine" and the progressive reduction in the degree of water fluoridation. Although most patients were followed up, this was only for a few years. Nevertheless, one case actually recurred during follow-up. Therefore, patients with OF should be encouraged to maintain follow-up for a longer period. PMID- 20026709 TI - Analysis of the masseter muscle in patients with systemic sclerosis: a study by magnetic resonance imaging. AB - OBJECTIVES: The aim of this study was to investigate the relationship between mandibular osteolysis and the T1 and T2 tissue signal strength and morphology of the masseter muscle in patients with systemic sclerosis (SS). METHODS: All patients with SS underwent panoramic radiography to detect mandibular osteolysis. MRI of the masseter was performed on 15 patients with SS and 10 healthy control subjects. RESULTS: All patients presented mean T1 values that were statistically equal. The patients with osteolysis presented mean T2 values that were the same as those of patients without osteolysis and normal individuals. The patients without osteolysis showed stronger T2 signals than did the normal individuals. The morphological analysis showed more atrophy, fatty replacement and rectified morphology in relation to the healthy subjects. CONCLUSIONS: The patients with SS presented more atrophy, fat replacement and rectification than did the normal individuals. The T2 signal was significantly stronger among the patients with SS without osteolysis than among normal individuals and among patients with SS and osteolysis. PMID- 20026710 TI - Periapical radiography and cone beam computed tomography for assessment of the periapical bone defect 1 week and 12 months after root-end resection. AB - OBJECTIVES: Our aim was to compare periapical radiography and cone beam CT (CBCT) for assessment of the periapical bone defect 1 week and 12 months after root-end resection. METHODS: 50 patients (58 teeth) with a persisting apical periodontitis in a root-filled tooth (incisor, canine or premolar) were treated with root-end resection. 1 week and 12 months post-operatively, a CBCT scan (NewTom 3G) and a periapical radiograph (Digora) were obtained. Three observers detected and measured the periapical bone defects on periapical radiographs and CBCT images (coronal and sagittal sections). RESULTS: 1 week post-operatively, a periapical bone defect area was measured in all teeth by all observers. The defect was 10% smaller on periapical radiographs (mean = 12.4 mm2, SD = 8.2) than on the CBCT images measured in the coronal plane (mean = 13.0 mm2, SD = 7.8), a difference which was not statistically significant (P = 0.58). 12 months post-operatively (n = 52), there was considerable variation between the observers' detection of a remaining defect on the periapical radiographs and the CBCT images. The average agreement between the periapical radiograph and the CBCT images in the coronal sections was 67%, and more defects were detected on CBCT than on periapical radiographs. CONCLUSIONS: On average, the periapical bone defect measured on periapical radiographs was approximately 10% smaller than on coronally sectioned CBCT images 1 week post-operatively. More remaining defects were detected 1 year after periapical surgery on CBCT images than on periapical radiographs, but it is uncertain how this information is related to success or failure after root-end resection. PMID- 20026711 TI - Effect of monitor display on detection of approximal caries lesions in digital radiographs. AB - OBJECTIVES: The aim was to compare the accuracy of five flat panel monitors for detection of approximal caries lesions. METHODS: Five flat panel monitors, Mermaid Ventura (15 inch, colour flat panel, 1024 x 768, 32 bit, analogue), Olorin VistaLine (19 inch, colour, 1280 x 1024, 32 bit, digital), Samsung SyncMaster 203B (20 inch, colour, 1024 x 768, 32 bit, analogue), Totoku ME251i (21 inch, greyscale, 1400 x 1024, 32 bit, digital) and Eizo FlexScan MX190 (19 inch, colour, 1280 x 1024, 32 bit, digital), were assessed. 160 approximal surfaces of human teeth were examined with a storage phosphor plate system (Digora FMX, Soredex) and assessed by seven observers for the presence of caries lesions. Microscopy of the teeth served as validation for the presence/absence of a lesion. RESULTS: The sensitivities varied between observers (range 7-25%) but the variation between the monitors was not large. The Samsung monitor obtained a significantly higher sensitivity than the Mermaid and Olorin monitors (P<0.02) and a lower specificity than the Eizo and Totoku monitors (P<0.05). There were no significant differences between any other monitors. The percentage of correct scores was highest for the Eizo monitor and significantly higher than for the Mermaid and Olorin monitors (P<0.03). CONCLUSIONS: There was no clear relationship between the diagnostic accuracy and the resolution or price of the monitor. The Eizo monitor was associated with the overall highest percentage of correct scores. The standard analogue flat panel monitor, Samsung, had higher sensitivity and lower specificity than some of the other monitors, but did not differ in overall accuracy for detection of carious lesions. PMID- 20026712 TI - Can dental pulp calcification serve as a diagnostic marker for carotid artery calcification in patients with renal diseases? AB - OBJECTIVES: The aim of this study was to determine the relationship between the presence of pulp calcification and carotid artery calcification on dental panoramic radiographs in end-stage renal disease (ESRD) patients on haemodialysis and renal transplant recipients. METHODS: A total of 60 nephrology patients (29 haemodialysis patients and 31 renal transplant recipients) participated in this study. For all patients, both panoramic and periapical radiographs were evaluated twice by three examiners to determine the presence or absence of narrowing of the dental pulps and pulp stones in pulp chambers and canals. The kappa coefficient was used for intervariable agreement, and interexaminer reliability was evaluated by the intraclass correlation coefficient. Panoramic radiographs were also evaluated to determine carotid calcification. Ultrasound examination confirmed radiographic findings of carotid calcification on panoramic radiographs. RESULTS: Carotid calcifications were detected in 11 patients (6 haemodialysis patients and 5 transplant recipients). 48 patients (22 haemodialysis patients, 26 transplanted recipients) had dental pulp narrowing, and 8 patients (5 haemodialysis patients, 3 transplant recipients) had pulp stones. There was no statistical relation between pulp narrowing and carotid artery calcification (CAC) in the haemodialysis patient group and renal transplant recipients. There was also no statistical relation between pulp stones and CAC in haemodialysis patients and renal transplant recipients. CONCLUSION: In our study, no relationship was found between the presence of pulpal calcification and CAC in ESRD patients on haemodialysis and renal transplant recipients. Therefore, the presence of pulp calcification does not seem to serve as a diagnostic marker for carotid atherosclerosis. PMID- 20026713 TI - Isolated submandibular metastasis from a contralateral thyroid papillary microcarcinoma: an unusual case. AB - Papillary carcinoma is the most common form of thyroid cancer. It is a relatively indolent disease, which commonly remains clinically silent until its incidental histological diagnosis in surgical material or at autopsy. A tumour less than 10 mm in size is termed a papillary microcarcinoma. Papillary microcarcinoma may present with clinical symptoms, most commonly jugulodigastric and pretracheal lymphadenopathy with or without palpable thyroid nodules. Isolated submandibular metastases are rare. We present the case of a submandibular metastasis arising from a solitary 3 mm papillary microcarcinoma of the thyroid on the contralateral side in a 46-year-old woman. We describe the ultrasound and MRI characteristics of the submandibular mass. The ultrasound findings in particular were suggestive of a thyroid malignancy and prompted detailed examination of the thyroid gland. Clinical and radiological examination of the thyroid was normal. To the best of our knowledge, we present the first report of a papillary microcarcinoma of the thyroid presenting as a contralateral and isolated submandibular mass. PMID- 20026714 TI - Sialographic findings in Wharton duct evagination. AB - Ductal evagination is a rare condition affecting the Wharton duct. The aim of this study was to establish the incidence, imaging features and clinical significance of ductal evagination in patients undergoing submandibular gland sialography. The sialographic findings and reports of 322 patients undergoing submandibular gland sialography during the period 1998-2007 were retrospectively reviewed. Ductal evagination was identified on sialograms as a unique diverticulum, filled with contrast medium, of the Wharton duct, with a narrow neck and a blind end. A ductal evagination was found in 5/322 patients with swelling and pain in the submandibular gland. It was always located in the middle tract of the Wharton duct. Other findings were: in 5/5 patients, an enlargement of the primary and secondary ducts due to sialodochitis; in 3/5 patients, salivary stones; in 1/5 patients, a sinuous distal tract of the Wharton duct; in 2/5 patients, a stenosis of the proximal tract of the Wharton duct. In our series, the incidence of wharton duct evagination was 1.6% incidence. It may represent a form of duct wall weakness although its cause is uncertain. It is, however, a condition that needs to be highlighted on sialograms for eventual inflammatory consequences or in guiding sialoendoscopy to avoid eventual ductal perforation. PMID- 20026718 TI - Celebrating nursing theory. PMID- 20026717 TI - Context-dependent pharmacology exhibited by negative allosteric modulators of metabotropic glutamate receptor 7. AB - Phenotypic studies of mice lacking metabotropic glutamate receptor subtype 7 (mGluR7) suggest that antagonists of this receptor may be promising for the treatment of central nervous system disorders such as anxiety and depression. Suzuki et al. (J Pharmacol Exp Ther 323:147-156, 2007) recently reported the in vitro characterization of a novel mGluR7 antagonist called 6-(4-methoxyphenyl)-5 methyl-3-(4-pyridinyl)-isoxazolo[ 4,5-c]pyridin-4(5H)-one (MMPIP), which noncompetitively inhibited the activity of orthosteric and allosteric agonists at mGluR7. We describe that MMPIP acts as a noncompetitive antagonist in calcium mobilization assays in cells coexpressing mGluR7 and the promiscuous G protein G alpha(15). Assessment of the activity of a small library of MMPIP-derived compounds using this assay reveals that, despite similar potencies, compounds exhibit differences in negative cooperativity for agonist-mediated calcium mobilization. Examination of the inhibitory activity of MMPIP and analogs using endogenous G(i/o)-coupled assay readouts indicates that the pharmacology of these ligands seems to be context-dependent, and MMPIP exhibits differences in negative cooperativity in certain cellular backgrounds. Electrophysiological studies reveal that, in contrast to the orthosteric antagonist (2S)-2-amino-2-[(1S,2S)-2 carboxyclycloprop-1-yl]-3-(xanth-9-yl) propanoic acid (LY341495), MMPIP is unable to block agonist-mediated responses at the Schaffer collateral-CA1 synapse, a location at which neurotransmission has been shown to be modulated by mGluR7 activity. Thus, MMPIP and related compounds differentially inhibit coupling of mGluR7 in different cellular backgrounds and may not antagonize the coupling of this receptor to native G(i/o) signaling pathways in all cellular contexts. The pharmacology of this compound represents a striking example of the potential for context-dependent blockade of receptor responses by negative allosteric modulators. PMID- 20026719 TI - Exploring the concept of power in a nursing context. PMID- 20026720 TI - Power, right, and truth: Foucault's triangle as a model for clinical power. AB - Power has historically been viewed from a position of dominance and authority. Using this lens leads one to a destiny wherein one individual or society has power over another. The power over approach is a hierarchical view, one that leads to someone else being oppressed, and one wherein the prevailing hegemony continues. If a different lens is used, wherein power does not reside within a position, but rather, within a person and within a relationship bounded by knowledge, a new destiny of power to and power of is created. These two approaches to power are examined and clinical power is offered as a lens that culminates in the understanding of power as a right and as truth imbedded with awareness and relationships. PMID- 20026721 TI - Research and the doctor of nursing practice: a cause for consternation. AB - This column concerns itself with the issue of research and the doctor of nursing practice. The reports of the Institute of Medicine about patient safety, quality in healthcare, and the restructuring of education of healthcare providers are reviewed. The reasons for the creation of the doctor of nursing practice are illuminated along with the essentials of educating nurses for the role and the position statement on nursing research by the American Association of Colleges of Nursing. Finally, the impact that those with a doctor of nursing practice who conduct nursing research may have on the discipline of nursing is considered. PMID- 20026722 TI - Nursing ethics and power in position. AB - Power in positions of authority and responsibility may be manifested in myriad ways in management positions, with all businesses and disciplines, including the discipline of nursing. From the nursing disciplinary theoretical perspective of humanbecoming, this column begins a discussion regarding the obligations of healthcare disciplines to address the ethical questions of what should and ought members of healthcare disciplines be doing to enhance the quality and integrity for healthcare recipients' experiences. Exercising power judiciously and with transparency will be defined and emphasized as important for members of healthcare disciplines, for preserving and protecting the legacy of future healthcare leaders. PMID- 20026723 TI - The power and possibility in listening. AB - In this column, the author describes the power of listening. There are three engaging listening modalities that are explored that form a foundation for exploring the power and possibility in listening: living true presence, participating in the council process; and engaging in strategic questioning. Several strategic questions are raised to explore the possibility of integrating these listening modalities into the nursing academy. PMID- 20026724 TI - Cocreating nursing practice: one organization at a time. PMID- 20026725 TI - Theories in action and how nursing practice changed. AB - Rogers' theoretical framework of diffusion of innovation guided the successful infusion of the educational training and implementation of the Magis model of care at a 570 bed hospital in the Chicagoland area. The Magis model of care was derived from several nursing theories along with information from the Institute of Family-Centered Care. By incorporating the components that relate to the institution's values and Magnet theme, the stages of innovation were readily adopted and sustained over the first year of implementation. The model has spread beyond the original and sister units as demonstrated by another department creating the Magnet poster with the various elements that they have incorporated into daily care delivery. What is so invigorating to the nursing administration is hearing how nursing staff articulates the care they give to the various components of the model and the theory that supports this practice. PMID- 20026726 TI - Exploring nursing perspectives on power. AB - In this column, the concept of power is introduced as basic to the nursing discipline, critical to practice and specific to nursing science as well as nursing practice. PMID- 20026727 TI - We're all here for the good of the patient: a dialogue on power. AB - Two nurse scholars, whose works have centered on power and related concepts, discuss the ideas that have shaped their concepts of power. In this conversation, they reflect on factors that constrain nurses and nursing in organizations, and consider some possibilities that could lead nurses to realize their full potential and influence in the healthcare system. PMID- 20026728 TI - Power as knowing participation in change: what's new and what's next. AB - The theory of power as knowing participation in change was derived from Rogers' science of unitary human beings. Following testing and research of the theory and measurement instrument, a practice methodology was developed and the health patterning practice model was initiated. Further developments of the theory are presented and future speculations are proposed. PMID- 20026729 TI - The universality of Rogers' Science of Unitary Human Beings. AB - The universality of Rogers' science is discussed within the context of science and religion with an emphasis on an alive universe. Barrett's power theory is presented as an exemplar of the universality of Rogers' science. The implications of an alive universe are discussed. Rogers and Barrett are recognized for their contributions to nursing science and the sacred alive universe. PMID- 20026730 TI - Power, trust, and Science of Unitary Human Beings influence political leadership: a celebration of Barrett's power theory. AB - The importance of nurses' participation in health policy leadership is discussed within the context of Rogers' science of unitary human beings, Barrett's power theory, and one nurse-politician's experience. Nurses have a major role to play in resolving public policy issues that influence the health of people. A brief review of the history of nurses in the political arena is presented. Research related to power and trust is reviewed. Suggested strategies for success in political situations are offered. PMID- 20026731 TI - Power, uncertainty, self-transcendence, and quality of life in breast cancer survivors. AB - The purpose of the study was to examine the relations among power, uncertainty, self-transcendence, and quality of life in breast cancer survivors from the perspective of Rogers' science of unitary human beings. A correlational, cross sectional study with purposive sampling (n = 104) was conducted. The results included statistically significant correlations, explained variance, and mediating relations among the pattern manifestations. The researcher concluded that there are complex and synergistic relations among the cluster of field pattern manifestations that contribute to quality of life in breast cancer survivors. Implications for theory, research, and practice are discussed. PMID- 20026732 TI - Evolutionary emergent: chief nurse executive as chief vision officer. AB - Barrett's power theory provides a theoretical framework that can be applied to the practice of nursing administration. From this perspective, the role of the system chief nurse executive can be seen as an evolutionary emergent, that of chief vision officer. Various organizational initiatives relevant to this role are discussed as exemplars of practice informed by the Barrett's power theory. PMID- 20026733 TI - Seeking to change while accepting the now. AB - The efforts of several nurse authors and Marsha Linehan to integrate diverse methods taken from philosophy, the human sciences, and spirituality with the biomedical model and natural science are discussed in relation to healthcare and nursing practice. PMID- 20026734 TI - The use of Hegelian dialectics in nursing science. AB - The aim of this column is to describe dialectics as a philosophy and method which can be used by nurses to make a contribution to nursing science. Dialectics can be used in three ways: as a philosophical approach, as a method using the dialectic laws, and as a method of describing the dialectic process by focusing on the dynamic elements of the process. Dialectics can also be combined with hermeneutics. PMID- 20026736 TI - Tumor microenvironments direct the recruitment and expansion of human Th17 cells. AB - Although Th17 cells play critical roles in the pathogenesis of many inflammatory and autoimmune diseases, their prevalence among tumor-infiltrating lymphocytes (TILs) and function in human tumor immunity remains largely unknown. We have recently demonstrated high percentages of Th17 cells in TILs from ovarian cancer patients, but the mechanisms of accumulation of these Th17 cells in the tumor microenvironment are still unclear. In this study, we further showed elevated Th17 cell populations in the TILs obtained from melanoma and breast and colon cancers, suggesting that development of tumor-infiltrating CD4(+) Th17 cells may be a general feature in cancer patients. We then demonstrated that tumor microenvironmental RANTES and MCP-1 secreted by tumor cells and tumor-derived fibroblasts mediate the recruitment of Th17 cells. In addition to their recruitment, we found that tumor cells and tumor-derived fibroblasts produce a proinflammatory cytokine milieu as well as provide cell-cell contact engagement that facilitates the generation and expansion of Th17 cells. We also showed that inflammatory TLR and nucleotide oligomerization binding domain 2 signaling promote the attraction and generation of Th17 cells induced by tumor cells and tumor-derived fibroblasts. These results identify Th17 cells as an important component of human TILs, demonstrate mechanisms involved in the recruitment and regulation of Th17 cells in tumor microenvironments, and provide new insights relevant for the development of novel cancer immunotherapeutic approaches. PMID- 20026737 TI - IL-15 prevents apoptosis, reverses innate and adaptive immune dysfunction, and improves survival in sepsis. AB - IL-15 is a pluripotent antiapoptotic cytokine that signals to cells of both the innate and adaptive immune system and is regarded as a highly promising immunomodulatory agent in cancer therapy. Sepsis is a lethal condition in which apoptosis-induced depletion of immune cells and subsequent immunosuppression are thought to contribute to morbidity and mortality. This study tested the ability of IL-15 to block apoptosis, prevent immunosuppression, and improve survival in sepsis. Mice were made septic using cecal ligation and puncture or Pseudomonas aeruginosa pneumonia. The experiments comprised a 2 x 2 full factorial design with surgical sepsis versus sham and IL-15 versus vehicle. In addition to survival studies, splenic cellularity, canonical markers of activation and proliferation, intracellular pro- and antiapoptotic Bcl-2 family protein expression, and markers of immune cell apoptosis were evaluated by flow cytometry. Cytokine production was examined both in plasma of treated mice and splenocytes that were stimulated ex vivo. IL-15 blocked sepsis-induced apoptosis of NK cells, dendritic cells, and CD8 T cells. IL-15 also decreased sepsis induced gut epithelial apoptosis. IL-15 therapy increased the abundance of antiapoptotic Bcl-2 while decreasing proapoptotic Bim and PUMA. IL-15 increased both circulating IFN-gamma, as well as the percentage of NK cells that produced IFN-gamma. Finally, IL-15 increased survival in both cecal ligation and puncture and P. aeruginosa pneumonia. In conclusion, IL-15 prevents two immunopathologic hallmarks of sepsis, namely, apoptosis and immunosuppression, and improves survival in two different models of sepsis. IL-15 represents a potentially novel therapy of this highly lethal disorder. PMID- 20026738 TI - A small, variable, and irregular killer cell Ig-like receptor locus accompanies the absence of MHC-C and MHC-G in gibbons. AB - The killer cell Ig-like receptors (KIRs) of NK cells recognize MHC class I ligands and function in placental reproduction and immune defense against pathogens. During the evolution of monkeys, great apes, and humans, an ancestral KIR3DL gene expanded to become a diverse and rapidly evolving gene family of four KIR lineages. Characterizing the KIR locus are three framework regions, defining two intervals of variable gene content. By analysis of four KIR haplotypes from two species of gibbon, we find that the smaller apes do not conform to these rules. Although diverse and irregular in structure, the gibbon haplotypes are unusually small, containing only two to five functional genes. Comparison with the predicted ancestral hominoid KIR haplotype indicates that modern gibbon KIR haplotypes were formed by a series of deletion events, which created new hybrid genes as well as eliminating ancestral genes. Of the three framework regions, only KIR3DL3 (lineage V), defining the 5' end of the KIR locus, is present and intact on all gibbon KIR haplotypes. KIR2DL4 (lineage I) defining the central framework region has been a major target for elimination or inactivation, correlating with the absence of its putative ligand, MHC-G, in gibbons. Similarly, the MHC-C-driven expansion of lineage III KIR genes in great apes has not occurred in gibbons because they lack MHC-C. Our results indicate that the selective forces shaping the size and organization of the gibbon KIR locus differed from those acting upon the KIR of other hominoid species. PMID- 20026739 TI - Early recycling compartment trafficking of CD1a is essential for its intersection and presentation of lipid antigens. AB - A major step in understanding differences in the nature of Ag presentation was the realization that MHC class I samples peptides transported to the endoplasmic reticulum from the cytosol, whereas MHC class II samples peptides from lysosomes. In contrast to MHC class I and II molecules that present protein Ags, CD1 molecules present lipid Ags for recognition by specific T cells. Each of the five members of the CD1 family (CD1a-e) localizes to a distinct subcompartment of endosomes. Accordingly, it has been widely assumed that the distinct trafficking of CD1 isoforms must also have evolved to enable them to sample lipid Ags that traffic via different routes. Among the CD1 isoforms, CD1a is unusual because it does not have a tyrosine-based cytoplasmic sorting motif and uniquely localizes to the early endocytic recycling compartment. This led us to predict that CD1a might have evolved to focus on lipids that localize to early endocytic/recycling compartments. Strikingly, we found that the glycolipid Ag sulfatide also localized almost exclusively to early endocytic and recycling compartments. Consistent with colocalization of CD1a and sulfatide, wild-type CD1a molecules efficiently presented sulfatide to CD1a-restricted, sulfatide-specific T cells. In contrast, CD1a:CD1b tail chimeras, that retain the same Ag-binding capacity as CD1a but traffic based on the cytoplasmic tail of CD1b to lysosomes, failed to present sulfatide efficiently. Thus, the intracellular trafficking route of CD1a is essential for efficient presentation of lipid Ags that traffic through the early endocytic and recycling pathways. PMID- 20026740 TI - Cutting edge: bortezomib-treated tumors sensitized to NK cell apoptosis paradoxically acquire resistance to antigen-specific T cells. AB - Bortezomib augments caspase-8 activity, rendering tumors susceptible to NK cell lysis. We hypothesized this effect would likewise sensitize tumors to Ag-specific CTLs. Instead, bortezomib-treated tumors that acquired sensitivity to NK cells simultaneously became resistant to killing by Ag-specific CTLs. Reduction in CTL killing persisted for days, was not due to changes in tumor expression of MHC class I, and was overcome by pulsing tumors with peptides recognized by tumor reactive CTLs. Tumor-outgrowth experiments showed tumors grew faster in SCID mice when cocultures of tumor-reactive CTLs and bortezomib-treated tumors were injected compared with untreated tumors (tumor doubling time 3.1 and 10.6 d, respectively; p < 0.01), whereas tumors grew slower in mice receiving cocultures of NK cells and bortezomib-treated tumors compared with untreated tumors (11.8 d and 5.0 d, respectively; p < 0.01). These findings demonstrate bortezomib-treated tumors sensitized to NK cell apoptosis paradoxically acquire resistance to CTLs as a consequence of bortezomib altering proteasomal processing and presentation of tumor Ags. PMID- 20026742 TI - Recruitment and differentiation of conventional dendritic cell precursors in tumors. AB - The origin of dendritic cells (DCs) in tumors remains obscure. Recent studies indicate that conventional DCs (cDCs) in lymphoid tissues arise from a distinct population of committed cDC precursors (pre-cDCs) that originate in bone marrow and migrate via blood. In this study, we show that pre-cDCs are precursors for cDCs in tumors. Pre-cDCs from tumors, bone marrow, and spleen exhibit similar morphologic, immunophenotypic, and functional properties. Adoptive transfer studies show that bone marrow pre-cDCs migrate from blood into the tumor where they generate cDCs. The chemokine CCL3, which is markedly upregulated in tumors, promotes pre-cDC recruitment. Both pre-cDCs and their cDC progeny actively proliferate within the tumor. cDCs that arise from pre-cDCs in tumors express lower levels of CD11c and MHC class II as compared with those in spleen; however, there was no difference in their abilities to respond to maturation stimuli or activate Ag-specific lymphocytes in vitro. Our study provides the first evidence supporting a role for pre-cDCs in DC development in tumors and suggests a potential target for cancer immunotherapy. PMID- 20026741 TI - Naturally occurring altered peptide ligands control Salmonella-specific CD4+ T cell proliferation, IFN-gamma production, and protective potency. AB - T cell activation required for host defense against infection is an intricately regulated and precisely controlled process. Although in vitro studies indicate that three distinct stimulatory signals are required for T cell activation, the precise contribution of each signal in regulating T cell proliferation and differentiation after in vivo infection is unknown. In this study, altered peptide ligands (APLs) derived from the protective Salmonella-specific FliC Ag and CD4+ T cells specific for the immune-dominant FliC(431-439) peptide within this Ag were used to determine how changes in TCR stimulation impact CD4+ T cell proliferation, differentiation, and protective potency. To explore the prevalence and potential use of altered TCR stimulation by bacterial pathogens, naturally occurring APLs containing single amino acid substitutions in putative TCR contact residues within the FliC(431-439) peptide were identified and used for stimulation under both noninfection and infection conditions. On the basis of this analysis, naturally-occurring APLs that prime proliferation of FliC-specific CD4+ T cells either more potently or less potently compared with the wild-type FliC(431-439) peptide were identified. Remarkably, despite these differences in proliferation, all of the APLs primed reduced IFN-gamma production by FliC(431 439)-specific CD4+ T cells after stimulation in vivo. Moreover, after expression of the parental FliC(431-439) peptide or each APL in recombinant Listeria monocytogenes, only CD4+ T cells stimulated with the wild-type FliC(431-439) peptide conferred significant protection against challenge with virulent Salmonella. These results reveal important and unanticipated roles for TCR stimulation in controlling pathogen-specific CD4+ T cell proliferation, differentiation, and protective potency. PMID- 20026743 TI - Lactic acid and acidification inhibit TNF secretion and glycolysis of human monocytes. AB - High concentrations of lactic acid (LA) are found under various pathophysiological conditions and are accompanied by an acidification of the environment. To study the impact of LA on TNF secretion, human LPS-stimulated monocytes were cultured with or without LA or the corresponding pH control. TNF secretion was significantly suppressed by low concentrations of LA (< or = 10 mM), whereas only strong acidification had a similar effect. This result was confirmed in a coculture model of human monocytes with multicellular tumor spheroids. Blocking synthesis of tumor-derived lactate by oxamic acid, an inhibitor of lactate dehydrogenase, reversed the suppression of TNF secretion in this coculture model. We then investigated possible mechanisms underlying the suppression. Uptake of [3-(13)C]lactate by monocytes was shown by hyphenated mass spectrometry. As lactate might interfere with glycolysis, the glycolytic flux of monocytes was determined. We added [1,2-(13)C(2)]glucose to the culture medium and measured glucose uptake and conversion into [2,3-(13)C(2)]lactate. Activation of monocytes increased the glycolytic flux and the secretion of lactate, whereas oxygen consumption was decreased. Addition of unlabeled LA resulted in a highly significant decrease in [2,3-(13)C(2)]lactate secretion, whereas a mere corresponding decrease in pH exerted a less pronounced effect. Both treatments increased intracellular [2,3-(13)C(2)]lactate levels. Blocking of glycolysis by 2 deoxyglucose strongly inhibited TNF secretion, whereas suppression of oxidative phosphorylation by rotenone had little effect. These results support the hypothesis that TNF secretion by human monocytes depends on glycolysis and suggest that LA and acidification may be involved in the suppression of TNF secretion in the tumor environment. PMID- 20026744 TI - Human plasmacytoid dendritic cells support Th17 cell effector function in response to TLR7 ligation. AB - Signals involved in the commitment of Th17 differentiation are of substantial interest for our understanding of antimicrobial defense mechanisms and autoimmune disorders. Various ways in which myeloid dendritic cells modulate Th17 differentiation have been identified. However, although plasmacytoid dendritic cells (PDCs) are regarded as important players in antiviral/antimicrobial host defense and autoimmune diseases, a putative modulatory role of PDCs in Th17 differentiation has not yet been elucidated in detail. We demonstrated that PDCs are capable of promoting Th17 differentiation in response to TLR7 stimulation. Further, both the differentiation of Th17 cells from naive T cells and the amplification of Th17 effector functions of memory T cells are promoted by PDCs after TLR7 activation. Our data are of strong clinical relevance because TLR7 activation in PDCs might represent one of the missing links between innate and adaptive immune mechanisms and contribute to the amplification of Th17-driven autoimmune disorders as well as viral host defense. PMID- 20026745 TI - IL-8 dictates glycosaminoglycan binding and stability of IL-18 in cystic fibrosis. AB - Dysregulation of airway inflammation contributes to lung disease in cystic fibrosis (CF). Inflammation is mediated by inflammatory cytokines, including IL 8, which illustrates an increase in biological half-life and proinflammatory activity when bound to glycosaminoglycans (GAGs). The aim of this project was to compare IL-8 and IL-18 for their relative stability, activity, and interaction with GAGs, including chondroitin sulfate, hyaluronic acid, and heparan sulfate, present in high quantities in the lungs of patients with CF. Bronchoalveolar lavage fluid was collected from patients with CF (n = 28), non-CF controls (n = 14), and patients with chronic obstructive pulmonary disease (n = 12). Increased levels of IL-8 and reduced concentrations of IL-18 were detected in bronchial samples obtained from CF individuals. The low level of IL-18 was not a defect in IL-18 production, as the pro- and mature forms of the molecule were expressed and produced by CF epithelial cells and monocytes. There was, however, a marked competition between IL-8 and IL-18 for binding to GAGs. A pronounced loss of IL 18 binding capacity occurred in the presence of IL-8, which displaced IL-18 from these anionic-matrices, rendering the cytokine susceptible to proteolytic degradation by neutrophil elastase. As a biological consequence of IL-18 degradation, reduced levels of IL-2 were secreted by Jurkat T lymphocytes. In conclusion, a novel mechanism has been identified highlighting the potential of IL-8 to determine the fate of other inflammatory molecules, such as IL-18, within the inflammatory milieu of the CF lung. PMID- 20026747 TI - Continuity in times of change. PMID- 20026748 TI - 2010: The Year of the Lung: an opportunity to improve lung health and fight lung disease. PMID- 20026746 TI - Constitutive activation of Wnt signaling favors generation of memory CD8 T cells. AB - T cell factor-1 (TCF-1) and lymphoid enhancer-binding factor 1, the effector transcription factors of the canonical Wnt pathway, are known to be critical for normal thymocyte development. However, it is largely unknown if it has a role in regulating mature T cell activation and T cell-mediated immune responses. In this study, we demonstrate that, like IL-7Ralpha and CD62L, TCF-1 and lymphoid enhancer-binding factor 1 exhibit dynamic expression changes during T cell responses, being highly expressed in naive T cells, downregulated in effector T cells, and upregulated again in memory T cells. Enforced expression of a p45 TCF 1 isoform limited the expansion of Ag-specific CD8 T cells in response to Listeria monocytogenes infection. However, when the p45 transgene was coupled with ectopic expression of stabilized beta-catenin, more Ag-specific memory CD8 T cells were generated, with enhanced ability to produce IL-2. Moreover, these memory CD8 T cells expanded to a larger number of secondary effectors and cleared bacteria faster when the immunized mice were rechallenged with virulent L. monocytogenes. Furthermore, in response to vaccinia virus or lymphocytic choriomeningitis virus infection, more Ag-specific memory CD8 T cells were generated in the presence of p45 and stabilized beta-catenin transgenes. Although activated Wnt signaling also resulted in larger numbers of Ag-specific memory CD4 T cells, their functional attributes and expansion after the secondary infection were not improved. Thus, constitutive activation of the canonical Wnt pathway favors memory CD8 T cell formation during initial immunization, resulting in enhanced immunity upon second encounter with the same pathogen. PMID- 20026749 TI - Nitric oxide synthase isoforms in lung parenchyma of patients with chronic obstructive pulmonary disease. PMID- 20026750 TI - What is autoimmunity and why is it likely to be important in chronic lung disease? PMID- 20026751 TI - Air pollution and pneumonia: the "old man" has a new "friend". PMID- 20026752 TI - Vitamin D and tuberculosis. PMID- 20026753 TI - Antimicrobial stewardship: There's no such thing as too much. PMID- 20026754 TI - Long-term outcomes of acute irritant-induced asthma and World Trade Center related lower airway disease. PMID- 20026756 TI - Stronger evidence for replication of NPPA using genome-wide genotyping data. PMID- 20026758 TI - Early life stress sensitizes rats to angiotensin II-induced hypertension and vascular inflammation in adult life. AB - Maternal separation during early life is an established chronic behavioral model of early life stress in rats. It is known that perinatal adverse environments increase activity of the renin-angiotensin (Ang) system, specifically Ang II, in adulthood. The aim of this study was to investigate whether the effects of early life stress augment the sensitivity of the Ang II pathway. Using Wistar Kyoto rats, the maternal separation (MS) protocol was performed by separating approximately half of the male pups from their mother 3 h/d from days 2 to 14 of life. Pups remaining with the mother at all times were used as controls. Maternal separation did not influence the plasma basal parameters, such as blood glucose, insulin, Ang II, Ang 1-7 and plasma renin activity. Furthermore, body weight, blood pressure, and heart rate were similar in MS and control rats. The acute pressor response to Ang II was not different in anesthetized MS and control rats. However, the chronic infusion of Ang II (65 ng/min SC) elicited an exaggerated hypertensive response in MS compared with control rats (P<0.05). Surprisingly, HR was dramatically increased during the second week of Ang II infusion in MS compared with control rats (P<0.05). This enhanced Ang II sensitivity was accompanied by a greater vascular inflammatory response in MS versus control rats. Chronic Ang II infusion increased vascular wall structure in both groups similarly. These data indicate that early life stress sensitizes rats to an increased hemodynamic and inflammatory response during Ang II-induced hypertension. PMID- 20026757 TI - New physiological concepts of the renin-angiotensin system from the investigation of precursors and products of angiotensin I metabolism. PMID- 20026759 TI - Effects of angiotensin metabolites in the coronary vascular bed of the spontaneously hypertensive rat: loss of angiotensin II type 2 receptor-mediated vasodilation. AB - Because angiotensin (Ang) metabolites mediate functions independent of Ang II, we investigated their effects on coronary flow in spontaneously hypertensive rats (SHRs). Results were compared with those in the iliac artery and abdominal aorta and the coronary circulation of the Wistar rat. Ang II, III, and IV decreased coronary flow in SHRs and Wistar rats, with Ang III and IV being approximately 10 and approximately 1000 times less potent than Ang II. Ang-(1-7) decreased coronary flow at concentrations >1 micromol/L in SHRs. The Ang II type 1 receptor antagonist irbesartan blocked the effects of Ang II, III, and IV, whereas the Ang II type 2 receptor antagonist PD123319 blocked the effects of Ang-(1-7). The maximal Ang II- and III-induced decreases in coronary flow in SHRs were twice as large as those in Wistar rats. PD123319 enhanced the constrictor effects of Ang II and III in Wistar rats so that, in the presence of this drug, their effects were comparable to those in SHRs. In contrast, PD123319 did not alter the Ang II- and III-induced responses in SHRs and blocked the constrictor effect of Ang II in iliac arteries. Ang II type 2 receptor-mediated relaxation did not occur in iliac arteries and abdominal aortas, and the constrictor effects of Ang metabolites in these vessels were identical in Wistar rats and SHRs. In conclusion, coronary constriction induced by Ang II, Ang III, and Ang-(1-7) is enhanced in SHRs as compared with Wistar rats. This is attributable to the absence of counterregulatory Ang II type 2 receptor-mediated relaxation and/or a change of the Ang II type 2 receptor phenotype from relaxant to constrictor. PMID- 20026760 TI - N-acetyl-seryl-aspartyl-lysyl-proline attenuates renal injury and dysfunction in hypertensive rats with reduced renal mass: council for high blood pressure research. AB - N-acetyl-seryl-aspartyl-lysyl-proline (Ac-SDKP) is a naturally occurring peptide of which the plasma concentration is increased 4- to 5-fold by angiotensin converting enzyme inhibitors. We reported previously that, in models of both hypertension and postmyocardial infarction, Ac-SDKP reduces cardiac inflammation and fibrosis. However, it is unknown whether Ac-SDKP can prevent or reverse renal injury and dysfunction in hypertension. In the present study, we tested the hypothesis that, in rats with 5/6 nephrectomy (5/6Nx)-induced hypertension, Ac SDKP reduces renal damage, albuminuria, and dysfunction by decreasing inflammatory cell infiltration and renal fibrosis and by increasing nephrin protein. Ac-SDKP (800 microg/kg per day, SC via osmotic minipump) or vehicle was either started 7 days before 5/6Nx (prevention) and continued for 3 weeks or started 3 weeks after 5/6Nx (reversal) and continued for another 3 weeks. Rats with 5/6Nx developed high blood pressure, left ventricular hypertrophy, albuminuria, decreased glomerular filtration rate, and increased macrophage infiltration (inflammation) and renal collagen content (fibrosis). Ac-SDKP did not affect blood pressure or left ventricular hypertrophy in either group; however, it significantly reduced albuminuria, renal inflammation, and fibrosis and improved glomerular filtration rate in both prevention and reversal groups. Moreover, slit diaphragm nephrin protein expression in the glomerular filtration barrier was significantly decreased in hypertensive rats. This effect was partially prevented or reversed by Ac-SDKP. We concluded that Ac-SDKP greatly attenuates albuminuria and renal fibrosis and improves renal function in rats with 5/6Nx. These effects may be related to decreased inflammation (macrophages) and increased nephrin protein. PMID- 20026761 TI - Inching towards a targeted therapy for preeclampsia. PMID- 20026762 TI - Going out on a LIM and cysteine-rich domains 1 limb: a new way to block calcineurin activity. PMID- 20026763 TI - Neointimal hyperplasia and vasoreactivity are controlled by genetic elements on rat chromosome 3. AB - Neointimal hyperplasia (NIH) can lead to restenosis after clinical vascular interventions. NIH results from complex and poorly understood interactions between signaling cascades in the extracellular matrix and the disrupted endothelium, which lead to vessel occlusion. Quantitative trait loci (QTLs) were reported previously on rat chromosomes 3 and 6 through linkage analysis of postinjury NIH in midiliac arterial sections. In the current study, substitution mapping validated the RNO3 NIH QTL but not the RNO6 NIH QTL. The SHR.BN3 congenic strain had a 3-fold increase in the percentage of NIH compared with the parental spontaneously hypertensive rat strain. A double congenic study of RNO3+RNO6 NIH QTL segments suggested less than additive effects of these 2 genomic regions. To test the hypothesis that changes in vessel dynamics account for the differences in NIH formation, we performed vascular reactivity studies in the Brown Norway (BN), spontaneously hypertensive rat (SHR), SHR.BN3, and SHR.BN6 strains. De endothelialized left common carotid artery rings of the SHR.BN3 showed an increased vascular responsiveness when treated with serotonin or prostaglandin F2(alpha), with significant differences in EC(50) and maximum effect (P<0.01) values compared with the spontaneously hypertensive rat parental strain. Because both vascular reactivity and percentage of NIH formation in the SHR.BN3 strain are significantly higher than the SHR strain, we postulate that these traits may be associated and are controlled by genetic elements on RNO3. In summary, these results confirm that the RNO3 NIH QTL carries the gene(s) contributing to postinjury NIH formation. PMID- 20026764 TI - Recombinant vascular endothelial growth factor 121 infusion lowers blood pressure and improves renal function in rats with placentalischemia-induced hypertension. AB - Antagonism of vascular endothelial growth factor (VEGF) signaling by soluble fms like tyrosine kinase 1 occurs during preeclampsia and is proposed to play an important role in the pathogenesis of preeclampsia. We reported recently that hypertension associated with chronic reductions in uteroplacental perfusion pressure (RUPP) is associated with increased soluble fms-like tyrosine kinase 1 and decreased free VEGF. Whether restoration of circulating VEGF can restore renal function and chronically decrease arterial pressure associated with placental ischemia remains unknown. We hypothesized that chronic infusion of VEGF(121) would attenuate hypertension, increase glomerular filtration rate, and reverse the endothelial dysfunction associated with chronic RUPP. VEGF(121) (at either 90 or 180 microg/kg per day) was administered for 5 days via osmotic minipump placed IP. Mean arterial pressure, renal function, and tissues were obtained on day 19 of pregnancy from RUPP+VEGF, RUPP, and normal pregnant dams. Mean arterial pressure was increased in the RUPP (131+/-3 mm Hg) compared with the normal pregnant (102+/-1 mm Hg) rats, and infusion of VEGF(121) resolved the hypertension (105+/-5 mm Hg). Glomerular filtration rate was decreased in the RUPP dams (1.5+/-0.3 mL/min) and restored to normal pregnant levels (3.1+/-0.5 mL/min) by VEGF(121) treatment (3.1+/-0.4 mL/min). Effective renal plasma flow, decreased by RUPP, was also increased by VEGF(121) infusion. Relaxation to acetylcholine was enhanced by the VEGF treatment (P<0.05). These data demonstrate that chronic infusion of VEGF(121) during late gestation restores glomerular filtration rate and endothelial function and reduces high blood pressure associated with placental ischemia. The present results suggest that VEGF(121) may be a candidate molecule for management of preeclampsia and its related complications. PMID- 20026765 TI - Blood pressure regulation in humans: calculation of an "error signal" in control of sympathetic nerve activity. AB - Within an individual, diastolic blood pressure (DBP) is negatively related to sympathetic burst incidence, such that lower pressure is associated with high burst incidence. Our goal was to explore the use of a calculation of a DBP "error signal" in the control of muscle sympathetic nerve activity in men and women. Baseline muscle sympathetic nerve activity was measured in healthy young men (n=22) and women (n=28). Women had significantly lower muscle sympathetic nerve activity than men (29+/-3 versus 43+/-2 bursts per 100 heartbeats; P<0.05). For each individual, the DBP at which there is a 50% likelihood of a muscle sympathetic nerve activity burst, the "T50" value, was calculated. Mean DBP was subtracted from the T50 blood pressure as an approximate error signal for burst activation. Error signal was negative in both sexes, indicating that DBP in both sexes was higher than the DBP value associated with a 50% burst likelihood. However, average error signal was significantly larger in women (-4+/-2 mm Hg) than in men (-1+/-0 mm Hg; P<0.05 versus women). We conclude that women operate at a mean DBP greater than their T50 compared with men, and this may be a contributing factor to low basal muscle sympathetic nerve activity in women. The relationship between error signal and burst incidence may provide important insight into the control of muscle sympathetic nerve activity across sexes and in various populations. PMID- 20026766 TI - Role of endothelin in mediating soluble fms-like tyrosine kinase 1-induced hypertension in pregnant rats. AB - Although soluble fms-like tyrosine kinase 1 (sFlt-1), an antagonist of vascular endothelial growth factor and placental growth factor, has been implicated in the pathogenesis of hypertension during preeclampsia, the mechanisms whereby enhanced sFlt-1 production leads to hypertension remain unclear. Both sFlt-1 and endothelin 1 productions are elevated in women with preeclampsia and in placental ischemic animal models of preeclampsia; however, the importance of endothelin 1 and sFlt-1 interactions in the control of blood pressure during pregnancy is unknown. The purpose of this study was to determine the role of endothelin 1 in mediating sFlt-1-induced hypertension in pregnant rats. To achieve this goal, sFlt-1 (3.7 microg/kg per day for 6 days) was infused into normal pregnant rats and pregnant rats treated with a selective endothelin type A receptor antagonist, ABT 627 (5 mg/kg per day for 6 days). Plasma concentration of sFlt-1 increased from 735+/-34 pg/mL in normal pregnant rats to 2498+/-645 pg/mL (P<0.05) with infusion of sFlt-1. Arterial pressure increased from 100+/-1 mm Hg in normal pregnant rats to 122+/-3 mm Hg (P<0.05) in sFlt-1-infused rats. Chronic increases in plasma sFlt-1 in normal pregnant rats increased preproendothelin mRNA expression in the renal cortices by approximately 3-fold. In addition, chronic endothelin type A receptor blockade completely abolished the blood pressure response to sFlt-1 in pregnant rats (104+/-3 versus 100+/-1 mm Hg; P<0.05), whereas the endothelin A receptor antagonist had no effect on arterial pressure in NP rats (105+/-2 versus 100+/-1 mm Hg). In conclusion, this study demonstrates that endothelin 1, via endothelin type A receptor activation, plays an important role in mediating the hypertension in response to excess sFlt-1 during pregnancy. PMID- 20026767 TI - Reactive oxygen species and cyclooxygenase 2-derived thromboxane A2 reduce angiotensin II type 2 receptor vasorelaxation in diabetic rat resistance arteries. AB - Angiotensin II has a key role in the control of resistance artery tone and local blood flow. Angiotensin II possesses 2 main receptors. Although angiotensin II type 1 receptor is well known and is involved in the vasoconstrictor and growth properties of angiotensin II, the role of the angiotensin II type 2 receptor (AT2R) remains much less understood. Although AT2R stimulation induces vasodilatation in normotensive rats, it induces vasoconstriction in pathological conditions involving oxidative stress and cyclooxygenase 2 expression. Thus, we studied the influence of cyclooxygenase 2 on AT2R-dependent tone in diabetes mellitus. Mesenteric resistance arteries were isolated from Zucker diabetic fatty (ZDF) and lean Zucker rats and studied using in vitro using wire myography. In ZDF rats, AT2R-induced dilation was lower than in lean rats (11% versus 21% dilation). Dilation in ZDF rats returned to the control (lean rats) level after acute superoxide reduction (Tempol and apocynin), cyclooxygenase 2 inhibition (NS398), or thromboxane A(2) synthesis inhibition (furegrelate). Cyclooxygenase 2 expression and superoxide production were significantly increased in ZDF rat arteries compared with arteries of lean rats. After chronic treatment with Tempol, AT2R-dependent dilation was equivalent in ZDF and lean rats. Chronic treatment of ZDF rats with NS398 also restored AT2R-dependent dilation to the control (lean rats) level. Plasma thromboxane B(2) (thromboxane A(2) metabolite), initially high in ZDF rats, was decreased by chronic Tempol and by chronic NS398 to the level found in lean Zucker rats. Thus, in type 2 diabetic rats, superoxide and thromboxane A(2) reduced AT2R-induced dilation. These findings are important to take into consideration when choosing vasoactive drugs for diabetic patients. PMID- 20026768 TI - Compliance, safety, and effectiveness of fixed-dose combinations of antihypertensive agents: a meta-analysis. AB - Two or more antihypertensive agents are increasingly used to control blood pressure (BP) in hypertensive patients. However, it is unclear whether fixed-dose combinations (FDCs) of 2 antihypertensive agents in a single tablet provide greater benefits than the corresponding free-drug components given separately. A meta-analysis was performed to assess compliance, persistence, BP control, and safety associated with FDCs in comparison with their free-drug components. Fifteen included studies (n=32331) reported on >or=1 of the evaluated outcomes. In 3 cohort studies and 2 trials reporting on drug compliance (n=17 999), the use of FDCs was associated with significantly better compliance (odds ratio: 1.21 [95% CI: 1.03 to 1.43]; P=0.02) compared with its corresponding free-drug combinations. In 3 cohort studies (n=12 653), there was a nonsignificant improvement in persistence with therapy (odds ratio: 1.54 [95% CI: 0.95 to 2.49]; P=0.08), and in 5 trials (n=1775) the odds ratio for adverse effects for FDC use compared with free-drug combination use was 0.80 (95% CI: 0.58 to 1.11; P=0.19). In 9 trials (n=1671) with BP data, use of an FDC was associated with nonsignificant changes in systolic and diastolic BPs of 4.1 mm Hg (95% CI: -9.8 to 1.5; P=0.15) and 3.1 mm Hg (95% CI: -7.1 to 0.9; P=0.13), respectively. In these BP-lowering comparisons, there was heterogeneity associated with differences in study design but no publication bias. In conclusion, compared with free-drug combinations, FDCs of antihypertensive agents are associated with a significant improvement in compliance and with nonsignificant beneficial trends in BP and adverse effects. PMID- 20026769 TI - LIM and cysteine-rich domains 1 regulates cardiac hypertrophy by targeting calcineurin/nuclear factor of activated T cells signaling. AB - LIM domain proteins are important regulators in cell growth, cell fate determination, cell differentiation, and remodeling of the cell cytoskeleton. LIM and cysteine-rich domains 1 (Lmcd1) is a novel protein that contain 2 LIM domains with regular spacing in the carboxy-terminal region. However, its roles in cardiac growth remain unknown. Here, we investigated whether Lmcd1 regulates cardiac hypertrophy in vitro and in vivo and elucidated the underlying molecular mechanisms. We used primary cultured cardiac myocytes and cardiac-specific Lmcd1 transgenic mice. In wild-type mice subjected to the aortic banding, cardiac hypertrophy was evident at 8 weeks. In transgenic mice, however, cardiac hypertrophy was significantly greater than that in wild-type mice, as estimated by heart weight:body weight ratio, cardiomyocyte area, and echocardiographic measurements, as well as cardiac atrial natriuretic peptide and B-type natriuretic peptide mRNA and protein levels. Our results further showed that cardiac fibrosis observed in wild-type aortic banding mice was augmented in transgenic aortic banding mice. Importantly, calcineurin activity and nuclear factor of activated T cells activation level were increased more in transgenic mice than those in wild-type mice after 8-week aortic banding. In vitro experiments in cardiac myocytes further revealed that angiotensin II-induced calcineurin activity and nuclear factor of activated T cells activation were enhanced by overexpression but blunted by downregulation of Lmcd1. In conclusion, our results suggest that Lmcd1 plays a critical role in the development of cardiac hypertrophy via activation of calcineurin/nuclear factor of activated T cells signaling pathway. PMID- 20026770 TI - Cost-effectiveness of prasugrel versus clopidogrel in patients with acute coronary syndromes and planned percutaneous coronary intervention: results from the trial to assess improvement in therapeutic outcomes by optimizing platelet inhibition with Prasugrel-Thrombolysis in Myocardial Infarction TRITON-TIMI 38. AB - BACKGROUND: In patients with acute coronary syndromes and planned percutaneous coronary intervention, the Trial to Assess Improvement in Therapeutic Outcomes by Optimizing Platelet Inhibition With Prasugrel-Thrombolysis in Myocardial Infarction 38 (TRITON-TIMI 38) demonstrated that treatment with prasugrel versus clopidogrel was associated with reduced rates of cardiovascular death, MI, or stroke and an increased risk of major bleeding. We evaluated the cost effectiveness of prasugrel versus clopidogrel from the perspective of the US healthcare system by using data from TRITON-TIMI 38. METHODS AND RESULTS: Detailed resource use data were prospectively collected for all patients recruited from 8 countries (United States, Australia, Canada, Germany, Italy, Spain, United Kingdom, and France; n=3373 prasugrel, n=3332 clopidogrel). Hospitalization costs were estimated on the basis of diagnosis-related group and in-hospital complications. Cardiovascular medication costs were estimated by using net wholesale prices (clopidogrel=$4.62/d; prasugrel=$5.45/d). Life expectancy was estimated from in-trial cardiovascular and bleeding events with the use of statistical models of long-term survival from a similar population from the Saskatchewan Health Database. Over a median follow-up of 14.7 months, average total costs (including study drug) were $221 per patient lower with prasugrel (95% confidence interval, -759 to 299), largely because of a lower rate of rehospitalization involving percutaneous coronary intervention. Prasugrel was associated with life expectancy gains of 0.102 years (95% confidence interval, 0.030 to 0.180), primarily because of the decreased rate of nonfatal MI. Thus, compared with clopidogrel, prasugrel was an economically dominant treatment strategy. If a hypothetical generic cost for clopidogrel of $1/d is used, the incremental net cost with prasugrel was $996 per patient, yielding an incremental cost-effectiveness ratio of $9727 per life-year gained. CONCLUSIONS: Among acute coronary syndrome patients with planned percutaneous coronary intervention, treatment with prasugrel versus clopidogrel for up to 15 months is an economically attractive treatment strategy. CLINICAL TRIAL REGISTRATION: clinicaltrials.gov. Unique identifier: NCT00097591. PMID- 20026771 TI - Natural history of very severe aortic stenosis. AB - BACKGROUND: We sought to assess the outcome of asymptomatic patients with very severe aortic stenosis. METHODS AND RESULTS: We prospectively followed 116 consecutive asymptomatic patients (57 women; age, 67 + or - 16 years) with very severe isolated aortic stenosis defined by a peak aortic jet velocity (AV-Vel) > or = 5.0 m/s (average AV-Vel, 5.37 + or - 0.35 m/s; valve area, 0.63 + or - 0.12 cm(2)). During a median follow-up of 41 months (interquartile range, 26 to 63 months), 96 events occurred (indication for aortic valve replacement, 90; cardiac deaths, 6). Event-free survival was 64%, 36%, 25%, 12%, and 3% at 1, 2, 3, 4, and 6 years, respectively. AV-Vel but not aortic valve area was shown to independently affect event-free survival. Patients with an AV-Vel > or = 5.5 m/s had an event-free survival of 44%, 25%, 11%, and 4% at 1, 2, 3, and 4 years, respectively, compared with 76%, 43%, 33%, and 17% for patients with an AV-Vel between 5.0 and 5.5 m/s (P<0.0001). Six cardiac deaths occurred in previously asymptomatic patients (sudden death, 1; congestive heart failure, 4; myocardial infarction, 1). Patients with an initial AV-Vel > or = 5.5 m/s had a higher likelihood (52%) of severe symptom onset (New York Heart Association or Canadian Cardiovascular Society class >II) than those with an AV-Vel between 5.0 and 5.5 m/s (27%; P=0.03). CONCLUSIONS: Despite being asymptomatic, patients with very severe aortic stenosis have a poor prognosis with a high event rate and a risk of rapid functional deterioration. Early elective valve replacement surgery should therefore be considered in these patients. PMID- 20026772 TI - Nitrite potently inhibits hypoxic and inflammatory pulmonary arterial hypertension and smooth muscle proliferation via xanthine oxidoreductase dependent nitric oxide generation. AB - BACKGROUND: Pulmonary arterial hypertension is a progressive proliferative vasculopathy of the small pulmonary arteries that is characterized by a primary failure of the endothelial nitric oxide and prostacyclin vasodilator pathways, coupled with dysregulated cellular proliferation. We have recently discovered that the endogenous anion salt nitrite is converted to nitric oxide in the setting of physiological and pathological hypoxia. Considering the fact that nitric oxide exhibits vasoprotective properties, we examined the effects of nitrite on experimental pulmonary arterial hypertension. METHODS AND RESULTS: We exposed mice and rats with hypoxia or monocrotaline-induced pulmonary arterial hypertension to low doses of nebulized nitrite (1.5 mg/min) 1 or 3 times a week. This dose minimally increased plasma and lung nitrite levels yet completely prevented or reversed pulmonary arterial hypertension and pathological right ventricular hypertrophy and failure. In vitro and in vivo studies revealed that nitrite in the lung was metabolized directly to nitric oxide in a process significantly enhanced under hypoxia and found to be dependent on the enzymatic action of xanthine oxidoreductase. Additionally, physiological levels of nitrite inhibited hypoxia-induced proliferation of cultured pulmonary artery smooth muscle cells via the nitric oxide-dependent induction of the cyclin-dependent kinase inhibitor p21(Waf1/Cip1). The therapeutic effect of nitrite on hypoxia induced pulmonary hypertension was significantly reduced in the p21-knockout mouse; however, nitrite still reduced pressures and right ventricular pathological remodeling, indicating the existence of p21-independent effects as well. CONCLUSIONS: These studies reveal a potent effect of inhaled nitrite that limits pathological pulmonary arterial hypertrophy and cellular proliferation in the setting of experimental pulmonary arterial hypertension. PMID- 20026773 TI - Histone deacetylase 3 is critical in endothelial survival and atherosclerosis development in response to disturbed flow. AB - BACKGROUND: Histone deacetylase 3 (HDAC3) is known to play a crucial role in the differentiation of endothelial progenitors. The role of HDAC3 in mature endothelial cells, however, is not well understood. Here, we investigated the function of HDAC3 in preserving endothelial integrity in areas of disturbed blood flow, ie, bifurcation areas prone to atherosclerosis development. METHODS AND RESULTS: En face staining of aortas from apolipoprotein E-knockout mice revealed increased expression of HDAC3, specifically in these branching areas in vivo, whereas rapid upregulation of HDAC3 protein was observed in endothelial cells exposed to disturbed flow in vitro. Interestingly, phosphorylation of HDAC3 at serine/threonine was observed in these cells, suggesting that disturbed flow leads to posttranscriptional modification and stabilization of the HDAC3 protein. Coimmunoprecipitation experiments showed that HDAC3 and Akt form a complex. Using a series of constructs harboring deletions, we found residues 136 to 206 of HDAC3 to be crucial in this interaction. Enforced expression of HDAC3 resulted in increased phosphorylation of Akt and upregulation of its kinase activity. In line with these findings, knockdown of HDAC3 with lentiviral vectors (shHDAC3) led to a dramatic decrease in cell survival accompanied by apoptosis in endothelial cells. In aortic isografts of apolipoprotein E-knockout mice treated with shHDAC3, a robust atherosclerotic lesion was formed. Surprisingly, 3 of the 8 mice that received shHDAC3-infected grafts died within 2 days after the operation. Miller staining of the isografts revealed disruption of the basement membrane and rupture of the vessel. CONCLUSIONS: Our findings demonstrated that HDAC3 serves as an essential prosurvival molecule with a critical role in maintaining the endothelial integrity via Akt activation and that severe atherosclerosis and vessel rupture in isografted vessels of apolipoprotein E knockout mice occur when HDAC3 is knocked down. PMID- 20026774 TI - Improved survival among patients with Eisenmenger syndrome receiving advanced therapy for pulmonary arterial hypertension. AB - BACKGROUND: Advanced therapy (AT) for pulmonary arterial hypertension in the context of congenital heart disease (Eisenmenger syndrome) improves pulmonary hemodynamics, functional class, and the 6-minute walk test. We examined the potential effect of AT on survival in this population. METHODS AND RESULTS: Data on all Eisenmenger patients attending our center over the past decade were collected. Survival rates were compared between patients on and off AT with the use of a modified version of the Cox model, which treats AT as a time-varying covariate. Baseline differences were adjusted for the use of propensity scores. A total of 229 patients (aged 34.5 + or - 12.6 years; 35.4% male) were included. The majority had complex anatomy, and 53.7% were in New York Heart Association class > or = III at baseline assessment. Mean resting saturations were 84.3%. Sixty-eight patients (29.7%) either were on AT or had AT initiated during follow up. During a median follow-up of 4.0 years, 52 patients died, only 2 of them while on AT. Patients on AT were at a significantly lower risk of death, both unadjusted and after adjustment for baseline clinical differences by propensity score regression adjustment (C statistic=0.80; hazard ratio, 0.16; 95% confidence interval, 0.04 to 0.71; P=0.015) and propensity score matching (hazard ratio, 0.10; 95% confidence interval, 0.01 to 0.78; P=0.028). CONCLUSIONS: AT for pulmonary arterial hypertension in a contemporary cohort of adults with Eisenmenger syndrome was associated with a lower risk of death. Survival benefits should be considered together with improved hemodynamics and functional class when decisions are made about AT in this population. PMID- 20026775 TI - Open heart surgery in patients with sickle cell hemoglobinopathy. AB - BACKGROUND: In patients with sickle cell trait or disease, reduced life expectancy and a tendency for complications are believed to negatively affect likelihood of survival after open heart surgery. The aim of this study was to review retrospectively the perioperative results of patients undergoing cardiac surgery at our institution. METHODS AND RESULTS: Between January 1995 and December 2006, 47 patients with either sickle cell disease or sickle cell trait underwent open heart surgery at our institution. The average age of the 29 male and 18 female patients was 20 years. Patient outcomes were analyzed through the use of the institutional database. Clinical and echocardiographic follow-up was complete in all patients except 3, with a mean follow-up period of 46 months. Current status could be confirmed in 32 patients. The most common operations included the treatment of congenital and valvular heart diseases. There were no coronary artery bypass grafting procedures. Average weight of the patients was 45 kg. Exchange transfusion was performed both preoperatively and during surgery. Mean preoperative hemoglobin S concentration was 30.4 + or - 3.2% and decreased to 8.1 + or - 2.6% while on pump. Average on-pump hematocrit value was 25.4 + or 3.7%; in the postoperative period, it increased to 32.7 + or - 4.9%. Mean cardiopulmonary bypass and cross-clamp times were 95 and 69 minutes, respectively. None of the patients had sickling crisis or acidosis. Postoperative complications included exploration for hemorrhage in 3 patients (6.4%), stroke in 2 patients (4.3%), renal failure in 2 patients (4.3%), and prolonged ventilation in 1 patient (2.1%). Average hospital stay was 8.3 days (range, 4 to 27 days). Early in-hospital death occurred in 1 patient (2.1%); currently, 31 patients (66%) remain alive and free of cardiac symptoms. CONCLUSIONS: Heart valve surgery and surgery for congenital heart diseases can be performed safely in patients with sickle cell disease or sickle cell trait with acceptable outcome and survival rates. PMID- 20026776 TI - Myocardial ischemia/reperfusion injury is mediated by leukocytic toll-like receptor-2 and reduced by systemic administration of a novel anti-toll-like receptor-2 antibody. AB - BACKGROUND: Reperfusion therapy for myocardial infarction is hampered by detrimental inflammatory responses partly via Toll-like receptor (TLR) activation. Targeting TLR signaling may optimize reperfusion therapy and enhance cell survival and heart function after myocardial infarction. Here, we evaluated the role of TLR2 as a therapeutic target using a novel monoclonal anti-TLR2 antibody. METHOD AND RESULTS: Mice underwent 30 minutes of ischemia followed by reperfusion. Compounds were administered 5 minutes before reperfusion. Cardiac function and dimensions were assessed at baseline and 28 days after infarction with 9.4-T mouse magnetic resonance imaging. Saline and IgG isotype treatment resulted in 34.5 + or - 3.3% and 31.4 + or - 2.7% infarction, respectively. Bone marrow transplantation experiments between wild-type and TLR2-null mice revealed that final infarct size is determined by circulating TLR2 expression. A single intravenous bolus injection of anti-TLR2 antibody reduced infarct size to 18.9 + or - 2.2% (P=0.001). Compared with saline-treated mice, anti-TLR2-treated mice exhibited less expansive remodeling (end-diastolic volume 68.2 + or - 2.5 versus 76.8 + or - 3.5 microL; P=0.046) and preserved systolic performance (ejection fraction 51.0 + or - 2.1% versus 39.9 + or - 2.2%, P=0.009; systolic wall thickening 3.3 + or - 6.0% versus 22.0 + or - 4.4%, P=0.038). Anti-TLR2 treatment significantly reduced neutrophil, macrophage, and T-lymphocyte infiltration. Furthermore, tumor necrosis factor-alpha, interleukin-1alpha, granulocyte macrophage colony-stimulating factor, and interleukin-10 were significantly reduced, as were phosphorylated c-jun N-terminal kinase, phosphorylated p38 mitogen-activated protein kinase, and caspase 3/7 activity levels. CONCLUSIONS: Circulating TLR2 expression mediates myocardial ischemia/reperfusion injury. Antagonizing TLR2 just 5 minutes before reperfusion reduces infarct size and preserves cardiac function and geometry. Anti-TLR2 therapy exerts its action by reducing leukocyte influx, cytokine production, and proapoptotic signaling. Hence, monoclonal anti-TLR2 antibody is a potential candidate as an adjunctive for reperfusion therapy in patients with myocardial infarction. PMID- 20026777 TI - Prognostic modeling of individual patient risk and mortality impact of ischemic and hemorrhagic complications: assessment from the Acute Catheterization and Urgent Intervention Triage Strategy trial. AB - BACKGROUND: Both ischemic and hemorrhagic complications increase mortality rate in acute coronary syndromes. Their frequency and relative importance vary according to individual patient risk profiles. We sought to develop prognostic models for the risk of myocardial infarction (MI) and major bleeding to assess their impact on risk of death and to examine the manner in which alternative antithrombotic regimens affect these risks in individual patients. METHODS AND RESULTS: The Acute Catheterization and Urgent Intervention Triage Strategy (ACUITY) trial randomized 13 819 patients with acute coronary syndrome to heparin plus a glycoprotein IIb/IIIa inhibitor, bivalirudin plus a glycoprotein IIb/IIIa inhibitor, or bivalirudin alone. By logistic regression, there were 5 independent predictors of MI within 30 days (n=705; 5.1%) and 8 independent predictors of major bleeding (n=645; 4.7%), only 2 of which were common to both event types. In a covariate-adjusted, time-updated Cox regression model, both MI and major bleeding significantly affected subsequent mortality rate (hazard ratios, 2.7 and 2.9, respectively; both P<0.001). Treatment with bivalirudin versus heparin plus a glycoprotein IIb/IIIa inhibitor was associated with a nonsignificant 8% increase in MI and a highly significant 50% decrease in major bleeding. Given the individual patient risk profiles and the fact that bivalirudin prevented approximately 6 major bleeds for each MI that might occur from its use, the estimated reduction in bleeding was greater than the estimated increase in MI by bivalirudin alone rather than heparin plus a glycoprotein IIb/IIIa inhibitor for nearly all patients. CONCLUSIONS: Consideration of the individual patient risk profile for MI and major bleeding and the relative treatment effects of alternative pharmacotherapies permits personalized decision making to optimize therapy of patients with acute coronary syndrome. CLINICAL TRIAL REGISTRATION: clinicaltrials.gov Identifier: NCT00093158. PMID- 20026778 TI - Relationship between cardiac rehabilitation and long-term risks of death and myocardial infarction among elderly Medicare beneficiaries. AB - BACKGROUND: For patients with coronary heart disease, exercise-based cardiac rehabilitation improves survival rate and has beneficial effects on risk factors for coronary artery disease. The relationship between the number of sessions attended and long-term outcomes is unknown. METHODS AND RESULTS: In a national 5% sample of Medicare beneficiaries, we identified 30 161 elderly patients who attended at least 1 cardiac rehabilitation session between January 1, 2000, and December 31, 2005. We used a Cox proportional hazards model to estimate the relationship between the number of sessions attended and death and myocardial infarction (MI) at 4 years. The cumulative number of sessions was a time dependent covariate. After adjustment for demographic characteristics, comorbid conditions, and subsequent hospitalization, patients who attended 36 sessions had a 14% lower risk of death (hazard ratio [HR], 0.86; 95% confidence interval [CI], 0.77 to 0.97) and a 12% lower risk of MI (HR, 0.88; 95% CI, 0.83 to 0.93) than those who attended 24 sessions; a 22% lower risk of death (HR, 0.78; 95% CI, 0.71 to 0.87) and a 23% lower risk of MI (HR, 0.77; 95% CI, 0.69 to 0.87) than those who attended 12 sessions; and a 47% lower risk of death (HR, 0.53; 95% CI, 0.48 to 0.59) and a 31% lower risk of MI (HR, 0.69; 95% CI, 0.58 to 0.81) than those who attended 1 session. CONCLUSIONS: Among Medicare beneficiaries, a strong dose response relationship existed between the number of cardiac rehabilitation sessions and long-term outcomes. Attending all 36 sessions reimbursed by Medicare was associated with lower risks of death and MI at 4 years compared with attending fewer sessions. PMID- 20026779 TI - Rosuvastatin in the prevention of stroke among men and women with elevated levels of C-reactive protein: justification for the Use of Statins in Prevention: an Intervention Trial Evaluating Rosuvastatin (JUPITER). AB - BACKGROUND: Prior primary prevention trials of statin therapy that used cholesterol criteria for enrollment have not reported significant decreases in stroke risk. We evaluated whether statin therapy might reduce stroke rates among individuals with low levels of cholesterol but elevated levels of high sensitivity C-reactive protein. METHODS AND RESULTS: In Justification for the Use of statins in Prevention: an Intervention Trial Evaluating Rosuvastatin (JUPITER), 17 802 apparently healthy men and women with low-density lipoprotein cholesterol levels <130 mg/dL and high-sensitivity C-reactive protein levels > or = 2.0 mg/L were randomly allocated to rosuvastatin 20 mg daily or placebo and then followed up for the occurrence of a first stroke. After a median follow-up of 1.9 years (maximum, 5.0 years), rosuvastatin resulted in a 48% reduction in the hazard of fatal and nonfatal stroke as compared with placebo (incidence rate, 0.18 and 0.34 per 100 person-years of observation, respectively; hazard ratio 0.52; 95% confidence interval, 0.34 to 0.79; P=0.002), a finding that was consistent across all examined subgroups. This finding was due to a 51% reduction in the rate of ischemic stroke (hazard ratio, 0.49; 95% confidence interval, 0.30 to 0.81; P=0.004), with no difference in the rates of hemorrhagic stroke between the active and placebo arms (hazard ratio, 0.67; 95% confidence interval, 0.24 to 1.88; P=0.44). CONCLUSIONS: Rosuvastatin reduces by more than half the incidence of ischemic stroke among men and women with low levels of low-density lipoprotein cholesterol levels who are at risk because of elevated levels of high-sensitivity C-reactive protein. CLINICAL TRIAL REGISTRATION: clinicaltrial.gov. Unique identifier: NCT00239681. PMID- 20026780 TI - Dispatcher-assisted cardiopulmonary resuscitation: risks for patients not in cardiac arrest. AB - BACKGROUND: Dispatcher-assisted cardiopulmonary resuscitation (CPR) instructions can increase bystander CPR and thereby increase the rate of survival from cardiac arrest. The risk of bystander CPR for patients not in arrest is uncertain and has implications for how assertive dispatch is in instructing CPR. We determined the frequency of dispatcher-assisted CPR for patients not in arrest and the frequency and severity of injury related to chest compressions. METHODS AND RESULTS: The investigation was a prospective cohort study of adult patients not in cardiac arrest for whom dispatchers provided CPR instructions in King County, Washington, between June 1, 2004, and January 31, 2007. The study focused on those who received chest compressions. Information was collected through review of the audio and written dispatch report, written emergency medical services report, hospital record, and telephone survey. Of the 1700 patients for whom dispatcher CPR instructions were initiated, 55% (938 of 1700) were in arrest, 45% (762 of 1700) were not in arrest, and 18% (313 of 1700) were not in arrest and received bystander chest compressions. Of the 247 not in arrest who received chest compressions and had complete outcome ascertainment, 12% (29 of 247) experienced discomfort, and 2% (6 of 247) sustained injuries likely or possibly caused by bystander CPR. Only 2% (5 of 247) suffered a fracture, and no patients suffered visceral organ injury. CONCLUSIONS: In this prospective study, the frequency of serious injury related to dispatcher-assisted bystander CPR among nonarrest patients was low. When coupled with the established benefits of bystander CPR among those with arrest, these results support an assertive program of dispatcher assisted CPR. PMID- 20026781 TI - Laboratory measures of exercise capacity and ventricular characteristics and function are weakly associated with functional health status after Fontan procedure. AB - BACKGROUND: Patients after the Fontan procedure are at risk for suboptimal functional health status, and associations with laboratory measures are important for planning interventions and outcome measures for clinical trials. METHODS AND RESULTS: Parents completed the generic Child Health Questionnaire for 511 Fontan Cross-Sectional Study patients 6 to 18 years of age (61% male). Associations of Child Health Questionnaire Physical and Psychosocial Functioning Summary Scores (FSS) with standardized measurements from prospective exercise testing, echocardiography, magnetic resonance imaging, and measurement of brain natriuretic peptide were determined by regression analyses. For exercise variables for maximal effort patients only, the final model showed that higher Physical FSS was associated only with higher maximum work rate, accounting for 9% of variation in Physical FSS. For echocardiography, lower Tei index (particularly for patients with extracardiac lateral tunnel connections), lower indexed end systolic volume, and the absence of atrioventricular valve regurgitation for patients having Fontan procedure at age <2 years were associated with higher Physical FSS, accounting for 14% of variation in Physical FSS. For magnetic resonance imaging, ratio of lower mass to end-diastolic volume and midquartiles of indexed end-systolic volume (nonlinear) were associated with higher Physical FSS, accounting for 11% of variation. Lower brain natriuretic peptide was significantly but weakly associated with higher Physical FSS (1% of variation). Significant associations for Psychosocial FSS with laboratory measures were fewer and weaker than for Physical FSS. CONCLUSIONS: In relatively healthy Fontan patients, laboratory measures account for a small proportion of the variation in functional health status and therefore may not be optimal surrogate end points for trials of therapeutic interventions. PMID- 20026782 TI - Endothelial-specific deletion of connexin40 promotes atherosclerosis by increasing CD73-dependent leukocyte adhesion. AB - BACKGROUND: Endothelial dysfunction is the initiating event of atherosclerosis. The expression of connexin40 (Cx40), an endothelial gap junction protein, is decreased during atherogenesis. In the present report, we sought to determine whether Cx40 contributes to the development of the disease. METHODS AND RESULTS: Mice with ubiquitous deletion of Cx40 are hypertensive, a risk factor for atherosclerosis. Consequently, we generated atherosclerosis-susceptible mice with endothelial-specific deletion of Cx40 (Cx40del mice). Cx40del mice were indeed not hypertensive. The progression of atherosclerosis was increased in Cx40del mice after 5 and 10 weeks of a high-cholesterol diet, and spontaneous lesions were observed in the aortic sinuses of young mice without such a diet. These lesions showed monocyte infiltration into the intima, increased expression of vascular cell adhesion molecule-1, and decreased expression of the ecto-enzyme CD73 in the endothelium. The proinflammatory phenotype of Cx40del mice was confirmed in another model of induced leukocyte recruitment from the lung microcirculation. Endothelial CD73 is known to induce antiadhesion signaling via the production of adenosine. We found that reducing Cx40 expression in vitro with small interfering RNA or antisense decreased CD73 expression and activity and increased leukocyte adhesion to mouse endothelial cells. These effects were reversed by an adenosine receptor agonist. CONCLUSIONS: Cx40-mediated gap junctional communication contributes to a quiescent nonactivated endothelium by propagating adenosine-evoked antiinflammatory signals between endothelial cells. Alteration in this mechanism by targeting Cx40 promotes leukocyte adhesion to the endothelium, thus accelerating atherosclerosis. PMID- 20026783 TI - Brain volume and metabolism in fetuses with congenital heart disease: evaluation with quantitative magnetic resonance imaging and spectroscopy. AB - BACKGROUND: Adverse neurodevelopmental outcome is an important source of morbidity in children with congenital heart disease (CHD). A significant proportion of newborns with complex CHD have abnormalities of brain size, structure, or function, which suggests that antenatal factors may contribute to childhood neurodevelopmental morbidity. METHODS AND RESULTS: Brain volume and metabolism were compared prospectively between 55 fetuses with CHD and 50 normal fetuses with the use of 3-dimensinal volumetric magnetic resonance imaging and proton magnetic resonance spectroscopy. Fetal intracranial cavity volume, cerebrospinal fluid volume, and total brain volume were measured by manual segmentation. Proton magnetic resonance spectroscopy was used to measure the cerebral N-acetyl aspartate: choline ratio (NAA:choline) and identify cerebral lactate. Complete fetal echocardiograms were performed. Gestational age at magnetic resonance imaging ranged from 25 1/7 to 37 1/7 weeks (median, 30 weeks). During the third trimester, there were progressive and significant declines in gestational age-adjusted total brain volume and intracranial cavity volume in CHD fetuses relative to controls. NAA:choline increased progressively over the third trimester in normal fetuses, but the rate of rise was significantly slower (P<0.001) in CHD fetuses. On multivariable analysis adjusted for gestational age and weight percentile, cardiac diagnosis and percentage of combined ventricular output through the aortic valve were independently associated with total brain volume. Independent predictors of lower NAA:choline included diagnosis, absence of antegrade aortic arch flow, and evidence of cerebral lactate (P<0.001). CONCLUSIONS: Third-trimester fetuses with some forms of CHD have smaller gestational age- and weight-adjusted total brain volumes than normal fetuses and evidence of impaired neuroaxonal development and metabolism. Hemodynamic factors may play an important role in this abnormal development. PMID- 20026785 TI - Endothelial-vasoprotective effects of high-density lipoprotein are impaired in patients with type 2 diabetes mellitus but are improved after extended-release niacin therapy. AB - BACKGROUND: High-density lipoprotein (HDL)-raising therapies are currently under intense evaluation, but the effects of HDL may be highly heterogeneous. We therefore compared the endothelial effects of HDL from healthy subjects and from patients with type 2 diabetes mellitus and low HDL (meeting the criteria for metabolic syndrome), who are frequently considered for HDL-raising therapies. Moreover, in diabetic patients, we examined the impact of extended-release (ER) niacin therapy on the endothelial effects of HDL. METHODS AND RESULTS: HDL was isolated from healthy subjects (n=10) and patients with type 2 diabetes (n=33) by sequential ultracentrifugation. Effects of HDL on endothelial nitric oxide and superoxide production were characterized by electron spin resonance spectroscopy analysis. Effects of HDL on endothelium-dependent vasodilation and early endothelial progenitor cell-mediated endothelial repair were examined. Patients with diabetes were randomized to a 3-month therapy with ER niacin (1500 mg/d) or placebo, and endothelial effects of HDL were characterized. HDL from healthy subjects stimulated endothelial nitric oxide production, reduced endothelial oxidant stress, and improved endothelium-dependent vasodilation and early endothelial progenitor cell-mediated endothelial repair. In contrast, these beneficial endothelial effects of HDL were not observed in HDL from diabetic patients, which suggests markedly impaired endothelial-protective properties of HDL. ER niacin therapy improved the capacity of HDL to stimulate endothelial nitric oxide, to reduce superoxide production, and to promote endothelial progenitor cell-mediated endothelial repair. Further measurements suggested increased lipid oxidation of HDL in diabetic patients, and a reduction after ER niacin therapy. CONCLUSIONS: HDL from patients with type 2 diabetes mellitus and metabolic syndrome has substantially impaired endothelial-protective effects compared with HDL from healthy subjects. ER niacin therapy not only increases HDL plasma levels but markedly improves endothelial-protective functions of HDL in these patients, which is potentially more important. CLINICAL TRIAL REGISTRATION: clinicaltrials.gov. Identifier: NCT00346970. PMID- 20026784 TI - Separating the mechanism-based and off-target actions of cholesteryl ester transfer protein inhibitors with CETP gene polymorphisms. AB - BACKGROUND: Cholesteryl ester transfer protein (CETP) inhibitors raise high density lipoprotein (HDL) cholesterol, but torcetrapib, the first-in-class inhibitor tested in a large outcome trial, caused an unexpected blood pressure elevation and increased cardiovascular events. Whether the hypertensive effect resulted from CETP inhibition or an off-target action of torcetrapib has been debated. We hypothesized that common single-nucleotide polymorphisms in the CETP gene could help distinguish mechanism-based from off-target actions of CETP inhibitors to inform on the validity of CETP as a therapeutic target. METHODS AND RESULTS: We compared the effect of CETP single-nucleotide polymorphisms and torcetrapib treatment on lipid fractions, blood pressure, and electrolytes in up to 67 687 individuals from genetic studies and 17 911 from randomized trials. CETP single-nucleotide polymorphisms and torcetrapib treatment reduced CETP activity and had a directionally concordant effect on 8 lipid and lipoprotein traits (total, low-density lipoprotein, and HDL cholesterol; HDL2; HDL3; apolipoproteins A-I and B; and triglycerides), with the genetic effect on HDL cholesterol (0.13 mmol/L, 95% confidence interval [CI] 0.11 to 0.14 mmol/L) being consistent with that expected of a 10-mg dose of torcetrapib (0.13 mmol/L, 95% CI 0.10 to 0.15). In trials, 60 mg of torcetrapib elevated systolic and diastolic blood pressure by 4.47 mm Hg (95% CI 4.10 to 4.84 mm Hg) and 2.08 mm Hg (95% CI 1.84 to 2.31 mm Hg), respectively. However, the effect of CETP single-nucleotide polymorphisms on systolic blood pressure (0.16 mm Hg, 95% CI -0.28 to 0.60 mm Hg) and diastolic blood pressure (-0.04 mm Hg, 95% CI -0.36 to 0.28 mm Hg) was null and significantly different from that expected of 10 mg of torcetrapib. CONCLUSIONS: Discordance in the effects of CETP single-nucleotide polymorphisms and torcetrapib treatment on blood pressure despite the concordant effects on lipids indicates the hypertensive action of torcetrapib is unlikely to be due to CETP inhibition or shared by chemically dissimilar CETP inhibitors. Genetic studies could find a place in drug-development programs as a new source of randomized evidence for drug-target validation in humans. PMID- 20026786 TI - Dispatcher-directed bystander initiated cardiopulmonary resuscitation: a safe step, but only a first step, in an integrated approach to improving sudden cardiac arrest survival. PMID- 20026787 TI - Balancing benefit and bleeding risk of antithrombotic agents in the individual patient with an acute coronary syndrome. PMID- 20026788 TI - Do more cardiac rehabilitation visits reduce events compared with fewer visits? PMID- 20026789 TI - Stroke due to undifferentiated aortic intimal sarcoma with disseminated metastatic lesions. PMID- 20026790 TI - Letter by Drinka regarding article, "Low-density lipoprotein cholesterol concentrations and death due to intraparenchymal hemorrhage: the Ibaraki Prefectural Health Study". PMID- 20026793 TI - Preparing effective grant applications. PMID- 20026792 TI - Palliative care in the treatment of advanced heart failure. PMID- 20026794 TI - Arrhythmogenic left ventricular cardiomyopathy. PMID- 20026795 TI - Breast cancer pathology report: in response to Zurrida et Al. PMID- 20026796 TI - "I am older, not elderly," said the patient with acute myeloid leukemia. PMID- 20026797 TI - Successful treatment with meloxicam, a cyclooxygenase-2 inhibitor, of patients with extra-abdominal desmoid tumors: a pilot study. PMID- 20026798 TI - Favorable prognostic impact of NPM1 mutations in older patients with cytogenetically normal de novo acute myeloid leukemia and associated gene- and microRNA-expression signatures: a Cancer and Leukemia Group B study. AB - PURPOSE: To analyze the prognostic significance of NPM1 mutations, and the associated gene- and microRNA-expression signatures in older patients with de novo, cytogenetically normal acute myeloid leukemia (CN-AML) treated with intensive chemotherapy. PATIENTS AND METHODS: One hundred forty-eight adults age >or= 60 years with de novo CN-AML, enrolled onto Cancer and Leukemia Group B protocols 9720 and 10201, were studied at diagnosis for NPM1, FLT3, CEBPA, and WT1 mutations, and gene- and microRNA-expression profiles. RESULTS: Patients with NPM1 mutations (56%) had higher complete remission (CR) rates (84% v 48%; P < .001) and longer disease-free survival (DFS; P = .047; 3-year rates, 23% v 10%) and overall survival (OS; P < .001; 3-year rates, 35% v 8%) than NPM1 wild-type patients. In multivariable analyses, NPM1 mutations remained independent predictors for higher CR rates (P < .001) and longer DFS (P = .004) and OS (P < .001), after adjustment for other prognostic clinical and molecular variables. Unexpectedly, the prognostic impact of NPM1 mutations was mainly observed in patients >or= 70 years. Gene- and microRNA-expression profiles associated with NPM1 mutations were similar across older patient age groups and similar to those in younger (< 60 years) patients with CN-AML. These profiles were characterized by upregulation of HOX genes and their embedded microRNAs and downregulation of the prognostically adverse MN1, BAALC, and ERG genes. CONCLUSION: NPM1 mutations have favorable prognostic impact in older patients with CN-AML, especially those age >or= 70 years. The gene- and microRNA-expression profiles suggest that NPM1 mutations constitute a marker defining a biologically homogeneous entity in CN AML that might be treated with specific and/or targeted therapies across age groups. PMID- 20026799 TI - Weighing risks and benefits of postchemotherapy retroperitoneal lymph node dissection: not so easy. PMID- 20026800 TI - Single-agent laromustine, a novel alkylating agent, has significant activity in older patients with previously untreated poor-risk acute myeloid leukemia. AB - PURPOSE An international phase II study of laromustine (VNP40101M), a sulfonylhydrazine alkylating agent, was conducted in patients age 60 years or older with previously untreated poor-risk acute myeloid leukemia (AML). PATIENTS AND METHODS Laromustine 600 mg/m(2) was administered as a single 60-minute intravenous infusion. Patients were age 70 years or older or 60 years or older with at least one additional risk factor-unfavorable AML karyotype, Eastern Cooperative Oncology Group (ECOG) performance status (PS) of 2, and/or cardiac, pulmonary, or hepatic comorbidities. Results Eighty-five patients (median age, 72 years; range, 60 to 87 years) were treated. Poor-risk features included age 70 years or older, 78%; adverse karyotype, 47%; PS of 2, 41%; pulmonary disease, 77%; cardiac disease, 73%; and hepatic disease, 3%. Ninety-six percent of patients had at least two risk factors, and 39% had at least four risk factors. The overall response rate (ORR) was 32%, with 20 patients (23%) achieving complete response (CR) and seven (8%) achieving CR with incomplete platelet recovery (CRp). ORR was 20% in patients with adverse cytogenetics; 32% in those age 70 years or older; 32% in those with PS of 2; 32% in patients with baseline pulmonary dysfunction; 34% in patients with baseline cardiac dysfunction; and 27% in 33 patients with at least four risk factors. Twelve (14%) patients died within 30 days of receiving laromustine therapy. Median overall survival was 3.2 months, with a 1-year survival of 21%; the median duration of survival for those who achieved CR/CRp was 12.4 months, with a 1-year survival of 52%. CONCLUSION Laromustine has significant single-agent activity in elderly patients with poor risk AML. Adverse events are predominantly myelosuppressive or respiratory. Response rates are consistent across a spectrum of poor-risk features. PMID- 20026801 TI - Phase I/II trial of metronomic chemotherapy with daily dalteparin and cyclophosphamide, twice-weekly methotrexate, and daily prednisone as therapy for metastatic breast cancer using vascular endothelial growth factor and soluble vascular endothelial growth factor receptor levels as markers of response. AB - PURPOSE Preclinical studies indicate that metronomic chemotherapy is antiangiogenic and synergistic with other antiangiogenic agents. We designed a phase I/II study to evaluate the safety and activity of adding dalteparin and prednisone to metronomic cyclophosphamide and methotrexate in women with measurable metastatic breast cancer (MBC). PATIENTS AND METHODS Patients received daily dalteparin and oral cyclophosphamide, twice-weekly methotrexate, and daily prednisone (dalCMP). The primary study end point was clinical benefit rate (CBR), a combination of complete response (CR), partial response (PR), and prolonged stable disease for > or = 24 weeks (pSD). Secondary end points included time to progression (TTP), duration of response, and overall survival (OS). Biomarker response to treatment was assessed by using plasma vascular endothelial growth factor (VEGF) and soluble VEGF receptors (sVEGFRs) -1 and -2. Results Forty-one eligible patients were accrued. Sixteen (39%) had no prior chemotherapy for MBC; 15 (37%) had two or more chemotherapy regimens for MBC. Toxicities were minimal except for transient grade 3 elevation of liver transaminases in 11 patients (27%) and grade 3 vomiting in one patient (2%). One patient (2%) had CR, six (15%) had PR, and three (7%) had pSD, for a CBR of 10 (24%) of 41 patients. Median TTP was 10 weeks (95% CI, 8 to 17 weeks), and median OS was 48 weeks (95% CI, 32 to 79 weeks). VEGF levels decreased but not significantly, whereas sVEGFR 1 and -2 levels increased significantly after 2 weeks of therapy. There was no correlation between response and VEGF, sVEGFR-1, or sVEGFR-2 levels. CONCLUSION Metronomic dalCMP is safe, well tolerated, and clinically active in MBC. PMID- 20026802 TI - Adenocarcinoma of colon presenting as Cushing's syndrome. PMID- 20026803 TI - Multicenter, phase II study of decitabine for the first-line treatment of older patients with acute myeloid leukemia. AB - PURPOSE: Older patients with acute myeloid leukemia (AML) have limited treatment options because of the lack of effectiveness and the toxicity of available therapies. We investigated the efficacy and toxicity of the hypomethylating agent decitabine as initial therapy in older patients with AML. PATIENTS AND METHODS: In this multicenter, phase II study, patients older than 60 years who had AML (ie, > 20% bone marrow blasts) and no prior therapy for AML were treated with decitabine 20 mg/m(2) intravenously for 5 consecutive days of a 4-week cycle. Response was assessed by weekly CBC and bone marrow biopsy after cycle 2 and after each subsequent cycle. Patients continued to receive decitabine until disease progression or an unacceptable adverse event occurred. RESULTS: Fifty five patients (mean age, 74 years) were enrolled and were treated with a median of three cycles (range, one to 25 cycles) of decitabine. The expert-reviewed overall response rate was 25% (complete response rate, 24%). The response rate was consistent across subgroups, including in patients with poor-risk cytogenetics and in those with a history of myelodysplastic syndrome. The overall median survival was 7.7 months, and the 30-day mortality rate was 7%. The most common toxicities were myelosuppression, febrile neutropenia, and fatigue. CONCLUSION: Decitabine given in a low-dose, 5-day regimen has activity as upfront therapy in older patients with AML, and it has acceptable toxicity and 30-day mortality. PMID- 20026804 TI - Azacitidine prolongs overall survival compared with conventional care regimens in elderly patients with low bone marrow blast count acute myeloid leukemia. AB - PURPOSE: In a phase III randomized trial, azacitidine significantly prolonged overall survival (OS) compared with conventional care regimens (CCRs) in patients with intermediate-2- and high-risk myelodysplastic syndromes. Approximately one third of these patients were classified as having acute myeloid leukemia (AML) under current WHO criteria. This analysis compared the effects of azacitidine versus CCR on OS in this subgroup. PATIENTS AND METHODS: Patients were randomly assigned to receive subcutaneous azacitidine 75 mg/m(2)/d or CCR (best supportive care [BSC] only, low-dose cytarabine (LDAC), or intensive chemotherapy [IC]). RESULTS: Of the 113 elderly patients (median age, 70 years) randomly assigned to receive azacitidine (n = 55) or CCR (n = 58; 47% BSC, 34% LDAC, 19% IC), 86% were considered unfit for IC. At a median follow-up of 20.1 months, median OS for azacitidine-treated patients was 24.5 months compared with 16.0 months for CCR treated patients (hazard ratio = 0.47; 95% CI, 0.28 to 0.79; P = .005), and 2 year OS rates were 50% and 16%, respectively (P = .001). Two-year OS rates were higher with azacitidine versus CCR in patients considered unfit for IC (P = .0003). Azacitidine was associated with fewer total days in hospital (P < .0001) than CCR. CONCLUSION: In older adult patients with low marrow blast count (20% to 30%) WHO-defined AML, azacitidine significantly prolongs OS and significantly improves several patient morbidity measures compared with CCR. PMID- 20026805 TI - Phase II study of clofarabine monotherapy in previously untreated older adults with acute myeloid leukemia and unfavorable prognostic factors. AB - PURPOSE: This phase II study assessed clofarabine monotherapy in older adults (>or= 60 years of age) with untreated acute myeloid leukemia (AML) and at least one unfavorable baseline prognostic factor. PATIENTS AND METHODS: Clofarabine was administered intravenously for 5 days at 30 mg/m(2)/d during induction and 20 mg/m(2)/d during reinduction/consolidation (six cycles maximum). The primary end point was overall remission rate (ORR; ie, complete remission [CR] plus CR with incomplete platelet recovery [CRp]). RESULTS: In 112 evaluable patients who were treated (median age, 71 years; range, 60 to 88 years), the ORR was 46% (38% CR, 8% CRp). ORR by unfavorable prognostic factor was 39% for patients >or= 70 years of age; 32% for Eastern Cooperative Oncology Group (ECOG) performance status 2; 51% for antecedent hematologic disorder; 54% for intermediate karyotype; 42% for unfavorable karyotype; and 48%, 51%, and 38% for one, two, and three risk factors, respectively. The median disease-free survival was 37 weeks (95% CI, 26 to 56 weeks). Median duration of remission was 56 weeks (95% CI, 33 to not estimable). The estimated median overall survival was 41 weeks (95% CI, 28 to 53 weeks) for all patients, 59 weeks for patients with CR/CRp, and 72 weeks for patients with CR. The 30-day all-cause mortality was 9.8%. The most common non laboratory drug-related toxicities (>or= 20% patients) were nausea, febrile neutropenia, vomiting, diarrhea, rash, and fatigue. CONCLUSION: Clofarabine is an active agent with acceptable toxicity in patients age 60 years or older with untreated AML who have at least one unfavorable prognostic factor. ORR did not seem affected by the presence of multiple unfavorable prognostic factors. PMID- 20026806 TI - Intensive loading dose of trastuzumab achieves higher-than-steady-state serum concentrations and is well tolerated. AB - PURPOSE Pharmacokinetics (PKs) and safety results from phase II/III trials suggest that, if high trastuzumab serum concentrations are reached early during treatment for human epidermal growth factor receptor 2 (HER2)-positive breast cancer, patients will gain clinical benefit, and the synergistic effects of trastuzumab and chemotherapy will be maximized. This phase I/II study evaluated the PKs, efficacy, and safety of a novel, intensive loading regimen of trastuzumab in women with HER2-positive metastatic breast cancer (MBC). PATIENTS AND METHODS An intensive loading regimen of trastuzumab was given (6 mg/kg intravenously on days 1, 8, and 15 followed by 6 mg/kg every 3 weeks from day 22) to women age 18 years or older with HER2-positive MBC who may have received previous surgery, radiotherapy, and/or chemotherapy. Study medication was continued until disease progression or withdrawal occurred. Results All eligible women (N = 72) received at least one dose of trastuzumab. Median estimated trough concentration of trastuzumab at the end of 3 weeks of the intensive loading regimen (total of 18 mg/kg of trastuzumab administered) of cycle 1 was 119 mg/L, which is higher than steady-state trough concentrations with a conventional weekly or every-3-week regimen (64.9 or 47.3 mg/L, respectively). No new or unexpected adverse events or increased cardiotoxicity were reported during the study. In patients with measurable disease (n = 47), response rate was 23.4%. Median time to progression was 7.7 months (in all patients). CONCLUSION An intensive loading regimen of trastuzumab achieved higher-than-steady-state serum concentrations during cycle 1, was well tolerated, and had a good efficacy profile. PMID- 20026807 TI - Management of disseminated nonseminomatous germ cell tumors with risk-based chemotherapy followed by response-guided postchemotherapy surgery. AB - PURPOSE: The management of patients with a radiographic complete response after chemotherapy remains controversial. The current study assesses the outcome for a modern, unselected patient population with disseminated testicular cancer with particular emphasis on those achieving a radiographic complete remission to combination chemotherapy. PATIENTS AND METHODS: All patients with disseminated nonseminoma seen between 1999 and 2007 at the British Columbia Cancer Agency (BCCA) as well as through the Oregon Testis Cancer Program were retrospectively reviewed. A total of 276 patients treated with combination chemotherapy were identified. A radiographic complete remission (CR) was defined as disappearance of all metastatic lesions or minimal residual tissue 200 g, OR = 2.1, 95% CI = 1.3 to 3.4; aristolochic acid: at 151-250 mg, OR = 1.4, 95% CI = 1.1 to 1.8, and at >500 mg, OR = 2.0, 95% CI = 1.4 to 2.9). A statistically significant linear dose response relationship was observed between the prescribed dose of Mu Tong or the estimated cumulative dose of aristolochic acid and the risk of urinary tract cancer (P < .001 for both). CONCLUSIONS: Consumption of aristolochic acid containing Chinese herbal products is associated with an increased risk of cancer of the urinary tract in a dose-dependent manner that is independent of arsenic exposure. PMID- 20026810 TI - Smoldering (asymptomatic) multiple myeloma: current diagnostic criteria, new predictors of outcome, and follow-up recommendations. AB - PURPOSE: To provide an overview on smoldering (asymptomatic) multiple myeloma (SMM) including current diagnostic criteria, predictors of progression, pattern of progression, and outcome. DESIGN: A comprehensive review of the literature on risk factors for progression, treatment attempts to delay progression and outcome in patients with SMM. RESULTS: The risk factors for progression of SMM include: plasma cell mass including M-protein size and percentage of bone marrow clonal plasma cells (BMPC), abnormal free light chain ratio, proportion of phenotypically abnormal BMPC, immunoparesis, evolution pattern (evolving v nonevolving), and pattern of magnetic resonance imaging abnormalities. Most patients with SMM progress with anemia and/or skeletal involvement. Immediate therapy with cytotoxic agents, such as melphalan/prednisone has not resulted in improved outcome. Patients should not be treated until progressive disease with end-organ damage occurs. Increasing anemia is the most reliable indicator of progression. CONCLUSION: These recently recognized predictors of outcome may be helpful for better disease monitoring and for investigation of new treatment approaches. Thus, recommendations for follow-up every to 3 to 6 months depending on the risk of progression are suggested, and clinical trials with new noncytotoxic biologically derived agents to delay progression, particularly in high-risk patients, are ongoing. PMID- 20026813 TI - E2F1 in melanoma progression and metastasis. AB - Metastases are responsible for cancer deaths, but the molecular alterations leading to tumor progression are unclear. Overexpression of the E2F1 transcription factor is common in high-grade tumors that are associated with poor patient survival. To investigate the association of enhanced E2F1 activity with aggressive phenotype, we performed a gene-specific silencing approach in a metastatic melanoma model. Knockdown of endogenous E2F1 via E2F1 small hairpin RNA (shRNA) expression increased E-cadherin expression of metastatic SK-Mel-147 melanoma cells and reduced their invasive potential but not their proliferative activity. Although growth rates of SK-Mel-147 and SK-Mel-103 xenograft tumors expressing E2F1 shRNA or control shRNA were similar, mice implanted with cells expressing E2F1 shRNA had a smaller area of metastases per lung than control mice (n = 3 mice per group; 5% vs 46%, difference = 41%, 95% confidence interval = 15% to 67%; P = .01; one-way analysis of variance). We identified epidermal growth factor receptor as a direct target of E2F1 and demonstrated that inhibition of receptor signaling abrogates E2F1-induced invasiveness, emphasizing the importance of the E2F1-epidermal growth factor receptor interaction as a driving force in melanoma progression that may serve as a paradigm for E2F1-induced metastasis in other human cancers. PMID- 20026812 TI - Familial coaggregation of cryptorchidism, hypospadias, and testicular germ cell cancer: a nationwide cohort study. AB - BACKGROUND: Cryptorchidism, hypospadias, and testicular germ cell cancer (TGCC) may be symptoms of a testicular dysgenesis syndrome that manifests during fetal life. To address the inheritability of this syndrome, we examined whether family history of cryptorchidism or hypospadias is associated with an increased risk of TGCC. METHODS: A total of 2,159,883 men born since 1953, identified through Danish health registers, were followed from April 2, 1968, through May 31, 2008. First-, second-, and third-degree relatives were identified in the Danish Family Relations Database; cryptorchidism and hypospadias patients were identified in the Danish Hospital Discharge Register; and TGCC patients were identified in the Danish Cancer Register. Poisson regression was used to calculate the risk ratio for TGCC by family history of cryptorchidism or hypospadias. RESULTS: A total of 5441 patients developed TGCC. A personal history of cryptorchidism or hypospadias was associated with an increased relative risk (RR) of developing TGCC (RR = 3.71, 95% confidence interval [CI] = 3.29 to 4.19; and RR = 2.13, 95% CI = 1.26 to 3.61, respectively). For example, in men in their thirties, the overall rate per 100 000 is 25.1 in the cohort, but 88.6 and 55.4 in men born with cryptorchidism or hypospadias, respectively. In contrast, relatives of a hypospadias patient did not have a statistically significantly increased risk of TGCC nor did the first- and second-degree relatives of cryptorchidism patients. However, we found a small increased risk of TGCC for third-degree relatives of patients with cryptorchidism. CONCLUSIONS: Having hypospadias or cryptorchidism was associated with an increased risk of developing TGCC. However, our finding that family history of hypospadias or cryptorchidism generally was not associated with increased risk of developing TGCC does not support the hypothesis of shared inheritability of cryptorchidism, hypospadias, and TGCC. PMID- 20026814 TI - Secondary tracheoesophageal puncture with in-office transnasal esophagoscopy. AB - OBJECTIVE: To evaluate the outcomes of voice restoration using office-based transnasal esophagoscopy (TNE) to guide placement of the secondary tracheoesophageal puncture (TEP). DESIGN: Retrospective chart review. SETTING: Two tertiary care medical centers. PATIENTS: The study included 39 patients who underwent the TNE-TEP procedure from January 2004 to December 2008. MAIN OUTCOME MEASURES: Clinical, demographic, and TE speech-related data were recorded to examine the ease, efficiency, complications, and speech-related outcomes. RESULTS: Among 39 patients identified, the average age was 65 years (age range, 47-83 years), with 32 male (82%) and 7 female (16%) patients. Twenty-five patients (64%) underwent total laryngectomy; 8 (21%) underwent total laryngectomy with partial pharyngectomy; and 14 (36%) underwent microvascular flap reconstruction. The overall success rate of secondary TNE-assisted TEP placement was 97% (n = 38), with 1 unsuccessful attempt. There was no statistically significant correlation found between patients having undergone radiation therapy (either before or after oncologic resection) or a cricopharyngeal myotomy and successful TEP placement, type of reconstruction used to close the pharyngeal defect when compared with the difficulty in the placement of the TEP, development of complications associated with TEP placement, use of the TEP prosthesis, or speech intelligibility at the last follow-up visit. Thirty-one patients (79%) were still using their TEP prosthesis for speech at the last follow-up visit. Of the patients reviewed, 28 (72%) had understandable TE speech. CONCLUSIONS: In office TNE-assisted TEP placement can safely be performed, with excellent speech outcomes. Reconstruction with musculocutaneous or microvascular free-tissue transfer did not limit our ability to place secondary TEPs with TNE. PMID- 20026815 TI - Intraoperative laryngeal nerve monitoring during thyroidectomy. AB - OBJECTIVE: To determine whether nerve integrity monitor testing during thyroidectomy predicts recurrent laryngeal nerve (RLN) function after surgery. DESIGN: Prospective cohort outcomes study PATIENTS: The study included 210 consecutive patients with thyroid abnormalities who underwent thyroidectomy. METHODS: All patients were intraoperatively monitored with a nerve integrity monitoring system (Xomed NIM II; Medtronic Inc, Fridley, Minnesota), and their vocal cord function was assessed with fiberoptic laryngoscopy before and after surgery. Normal and impaired vocal cord function were compared using an independent t test with respect to postoperative vocal cord mobility, length of the RLN dissection, and the minimum stimulus needed to generate a response at the completion of surgery. RESULTS: There was a statistically significant difference between the stimulus in milliamperes required to stimulate normal vs abnormal functioning nerves at the completion of the procedure at the cricoarytenoid joint (P = .02) and at the distal end of the RLN dissection (P < .01). A greater length of dissected nerve was associated with normal vocal cord function; however, it was not statistically significant (P = .07). CONCLUSION: These data suggest that an RLN that responds at lower-intensity stimulation ( .99). The cerebrospinal fluid leak resolved spontaneously in 3 patients, and the fourth patient underwent successful endoscopic repair. With a mean follow-up of 37 months, 18 patients (15%) experienced local recurrence, with a local disease control of 85%. Regional and distant failure occurred as the first sign of disease recurrence in 6% and 5% of patients, respectively. The 5- and 10-year disease-specific survival rates were 87% and 80%, respectively. Disease recurrence and survival did not differ significantly between the EEA group and the CEA group. CONCLUSIONS: To the best of our knowledge, this is the largest US series to date of patients with malignant tumors of the sinonasal tract treated with endoscopic resection. Our results suggest that, in well-selected patients and with appropriate use of adjuvant therapy, endoscopic resection of sinonasal cancer results in acceptable oncologic outcomes. PMID- 20026820 TI - Oropharyngeal cancer: a case for single modality treatment with transoral laser microsurgery. AB - OBJECTIVE: To demonstrate the role of transoral laser microsurgery (TLM) in the treatment of oropharyngeal cancer. DESIGN: A 2-center retrospective case series analysis. SETTING: Two tertiary care medical centers. PATIENTS: The study population comprised 69 patients with previously untreated select T1 to T3, N0 to N2 squamous cell carcinoma of the oropharynx, of whom 44 (74%) had no indication for adjuvant RT and 25 (36%) had an indication for adjuvant RT to the neck alone but declined radiotherapy. The primary tumor sites were the tonsil (n = 28 [41%]), tongue base (n = 28 [41%]), pharyngeal wall (n = 8 [12%]), soft palate (n = 4 [6%]), and vallecula (n = 1 [1%]). INTERVENTIONS: Transoral laser microsurgery in 69 patients, with neck dissection in 59 patients (83%). MAIN OUTCOME MEASURES: Complications, local and regional control, overall and disease specific survival, swallow function, and feeding tube dependence. RESULTS: Over the mean follow-up period of 44 months, 66 of 69 patients had no disease recurrence at the primary site. The 5-year local control estimate was 94%. The mean duration of hospitalization was 3 days. There were no major complications relating to TLM. No patient required a permanent feeding or tracheostomy tube. For stage I, II, and III disease, the 5-year Kaplan-Meier estimates of locoregional control were 90%, 73%, and 70%, respectively. The 5-year overall survival estimate was 86%. CONCLUSIONS: Transoral laser microsurgery alone with or without neck dissection is an effective approach for select T1 to T3, N0, or N1 oropharyngeal cancer. Low levels of morbidity, short treatment duration, and excellent disease control make it an attractive therapeutic strategy. The treatment option of endoscopic-assisted laser microsurgery should be discussed by the multidisciplinary team for patients presenting with tumors suitable for this approach. PMID- 20026821 TI - Transoral resection for squamous cell carcinoma of the base of the tongue. AB - OBJECTIVE: To review the oncologic and functional outcomes of patients with squamous cell carcinoma of the base of the tongue who underwent transoral tumor resection and neck dissection with or without postoperative adjuvant therapy. DESIGN: Retrospective medical record review. SETTING: Tertiary referral center. PATIENTS: All patients undergoing transoral resection of squamous cell carcinoma on the base of the tongue as part of their primary treatment from January 1, 1996, through January 31, 2005. MAIN OUTCOME MEASURES: We analyzed overall survival, disease-specific survival, local control, and locoregional control rates using the Kaplan-Meier method. Speech and swallowing function and treatment related morbidity were also analyzed. RESULTS: A total of 20 patients underwent transoral resection. Four patients had surgery only, 12 had surgery and radiotherapy, and 4 had surgery and chemoradiotherapy. One patient had stage II disease, 3 had stage III disease, and 16 had stage IVA disease. The Kaplan-Meier overall survival rate was 90.0%, and the disease-specific survival rate was 94.7% at 2 years, with a mean follow-up of 3.7 years. Median hospital stay was 4.7 days. Patients who received a tracheostomy underwent decannulation with a median tracheostomy time of 5.5 days. Seven of 9 patients who received a percutaneous endoscopic gastrostomy tube had it removed. Three patients developed local recurrence, there were no regional recurrences, and 2 patients developed distant metastasis. CONCLUSIONS: Transoral resection of squamous cell carcinoma of the base of the tongue with postoperative adjuvant therapy provided excellent local and regional control and minimized morbidity. Transoral resection is a reasonable treatment option for patients with oropharyngeal squamous cell carcinoma, resulting in very low overall loss of organ function in properly selected patients. PMID- 20026822 TI - Location of bacterial biofilm in the mucus overlying the adenoid by light microscopy. AB - OBJECTIVE: To determine the location of bacteria and biofilm in adenoid tissue and in mucus overlying the adenoid. DESIGN: Adenoids removed in 1 piece were oriented to the cephalic and caudal ends. Mucus was fixed by the gradual addition of Carnoy fluid. Consecutive histologic sections were stained with periodic acid Schiff for visualization of the exopolysaccharide matrix, Giemsa for visualization of bacteria and cells, and fluorescent in situ hybridization with a universal probe for visualization of bacteria. SETTING: Department of Otolaryngology-Head and Neck Surgery, University of Virginia. PARTICIPANTS: We obtained adenoids from children 10 years or younger who had chronic adenotonsillitis or obstructive sleep apnea. Twenty-seven adenoids were used to develop the fixation method. We examined histologic sections from 9 of 10 adenoids fixed using the final fixation protocol. One adenoid that was missing the surface epithelium was excluded from further evaluation. MAIN OUTCOME MEASURE: Identification of bacteria by light microscopy. RESULTS: Bacteria in large numbers were present in the mucus overlying the surface of all 9 adenoids; bacteria were not found in the parenchyma of the adenoids below the epithelial surface. Bacterial biofilms were present on 8 of the 9 adenoids. Sessile (attached) biofilm was present on the caudal end of only 1 adenoid. Multiple planktonic (unattached) biofilms were present on 7 adenoids, always in areas not subject to mucus flow. Biofilms were most common on the caudal portions of adenoids. CONCLUSIONS: Bacteria of the adenoid reside in secretions on the surface and in crypts. Biofilms, predominantly planktonic, were present on 8 of 9 adenoids excised because of hypertrophy. Whether biofilms have a role in the causation of adenoid hypertrophy is not known. PMID- 20026823 TI - Diagnostic value of nasal allergen challenge combined with radiography and ultrasonography in chronic maxillary sinus disease. AB - OBJECTIVE: To investigate the possible role of nasal allergy in chronic disease of the maxillary sinuses (CDMS) by means of nasal provocation test (NPT) with allergen combined with radiography and ultrasonography. DESIGN: Prospective clinical controlled study. SETTING: Academic referral center. PATIENTS: Seventy one patients with CDMS and 16 control subjects with allergic rhinitis but no history of sinus disease. INTERVENTIONS: In the 71 patients, a total of 135 NPTs and 71 control challenges with phosphate-buffered saline were performed by rhinomanometry combined with radiography and ultrasonography. In the control patients, 16 positive NPTs were repeated and combined with radiography and ultrasonography. MAIN OUTCOME MEASURES: Number, type, and timing of nasal responses with accompanying changes on radiographs and ultrasonograms. RESULTS: Of the 71 patients, 67 developed 104 positive nasal responses of various types (P < .001), 89 of which were accompanied by significant changes on radiographs (P = .008), whereas 83 were also associated with significant changes on ultrasonograms (P = .007). No significant changes on the radiographs or the ultrasonograms were recorded during the 71 phosphate-buffered saline control tests in the patients with CDMS (P = .14 and .06, respectively) or during the 16 NPTs in control subjects (P = .15 and .12, respectively). The radiographic and ultrasonographic findings were significantly correlated (r = 0.81; P < .01). CONCLUSIONS: Nasal allergy may be involved in some patients with CDMS, resulting in appearance of a maxillary sinus response. Monitoring this response by means of serial ultrasonography and, if necessary, also by conventional radiography or computed tomography simultaneously with the nasal challenge with allergen seems to be a very useful diagnostic supplement allowing additional therapeutic measures focused on the nasal allergy. PMID- 20026824 TI - Cutting and suture technique of the caudal septal cartilage for the management of caudal septal deviation. AB - OBJECTIVES: To introduce the cutting and suture technique of the caudal L-strut for the management of caudal septal deviation and to evaluate its efficacy and surgical outcomes. DESIGN: Retrospective study. SETTING: Tertiary care rhinology clinic. PATIENTS: Forty-five patients who underwent endonasal septoplasty using the cutting and suture technique of the caudal L-strut. INTERVENTIONS: After elevation of the mucoperichondrial flap, deviated portions of cartilage and bone were excised, leaving at least a 1.5-cm strip of L-strut. If caudal septal deviation persisted, the caudal strut was cut at the convex-most part, and the cut ends were slightly overlapped and sutured together. MAIN OUTCOME MEASURES: Improvement in the treatment of nasal obstruction using a visual analog scale and a questionnaire for subjective satisfaction were evaluated 2 to 6 months after septoplasty. To evaluate outcomes objectively, endoscopic photographs of the nasal cavity before and after surgery were evaluated by 2 independent surgeons. RESULTS: Significant improvement in the treatment of nasal obstruction was achieved, with mean visual analog scale scores of 7.93 preoperatively and 3.63 postoperatively (P < .001). Subjective satisfaction was rated as much improved in 68% of patients, improved in 15%, and no change in 17%. Endoscopic examinations showed that 51% of patients had near-complete correction of the septum and that 47% had improved but a little persisting caudal deviation. One patient had no change in caudal septal deviation on endoscopic examination. CONCLUSION: The cutting and suture technique of the caudal L-strut seems to be a useful technique that can be performed with relative ease and simplicity. PMID- 20026825 TI - The histologic relationship of preauricular sinuses to auricular cartilage. AB - OBJECTIVE: To determine the histologic relationship and distance between excised preauricular epithelial sinus tract and the adjacent auricular cartilage (sinocartilaginous distance) in a series of patients. The excision of preauricular sinuses is a common surgical procedure. Recurrences are frequent and can be technically challenging. While advocated by several authors, the surgical removal of adjacent auricular cartilage is not universally performed. DESIGN: Retrospective case series. SETTING: Children's Hospital of Philadelphia. PATIENTS: Fifty-two pediatric patients who underwent surgical excision of preauricular sinus tracts and adjacent auricular cartilage. INTERVENTIONS: Between September 1, 2005, and July 31, 2007, the preauricular sinus tracts and adjacent auricular cartilage were excised from 52 pediatric patients. A pathologist reviewed a total of 58 specimens to determine the relationship between epithelial tract and cartilage. MAIN OUTCOME MEASURE: The sinocartilaginous distance in microns. RESULTS: Patient ages ranged from 8 months to 17 years (mean age, 4 years). In all but 1 case, the tracts were in close proximity to the cartilage. The average sinocartilaginous distance was 472 mum (median distance, 400 mum); the 25th percentile was 250 mum. In over 50% of the specimens, the sinocartilaginous distance was less than 0.5 mm, and in nearly all of the these, the epithelial tract was in continuity with stromal tissue histologically indistinguishable from perichondrium. CONCLUSIONS: The observed sinocartilaginous distances suggest that it may be difficult to dissect most sinus tracts from the cartilage. The routine removal of a small portion of auricular cartilage along with the sinus tract may yield a more thorough excision and help to prevent recurrence. PMID- 20026826 TI - Surgery as a teachable moment: lost opportunities to improve public health. PMID- 20026827 TI - Impact of advancing age on abdominal surgical outcomes. AB - OBJECTIVE: To describe the population-level risk of adverse outcomes among older adults undergoing common abdominal surgical procedures. DESIGN: Retrospective, population-based cohort study. SETTING: Washington State hospital discharge database. PARTICIPANTS: A total of 101 318 adults 65 years or older who underwent common abdominal procedures such as cholecystectomy, colectomy, and hysterectomy from 1987 through 2004. MAIN OUTCOME MEASURES: Ninety-day rates of postsurgical morbidity and mortality. RESULTS: The 90-day cumulative incidence of complications was 17.3%, with a 90-day mortality rate of 5.4%. Advancing age was associated with increasing frequency of complications (65-69 years, 14.6%; 70-74 years, 16.1%; 75-79 years, 18.8%; 80-84 years, 19.9%; 85-89 years, 22.6%; and >or=90 years, 22.7%; trend test, P < .001) and mortality (65-69 years, 2.5%; 70 74 years, 3.8%; 75-79 years, 6.0%; 80-84 years, 8.1%; 85-89 years, 12.6%; and >or=90 years, 16.7%; trend test, P < .001). After adjusting for demographic, patient, and surgical characteristics as well as hospital volume, the odds of early postoperative death increased considerably with each advance in age category. These associations were found among patients with both cancer and noncancer diagnoses and for both elective and nonelective admissions (trend test, P < .001). CONCLUSIONS: Among older adults, the risk of complications and early death after commonly performed abdominal procedures is greater than previously reported. These rates should be considered in ongoing quality improvement initiatives and may be helpful when counseling patients regarding abdominal operations. PMID- 20026828 TI - Colon cancer and low lymph node count: who is to blame? AB - OBJECTIVE: To identify the factors that contribute to the disparity in the number of lymph nodes examined for curative colon cancer resections. DESIGN: Our prospectively accrued cancer registry was analyzed for all colon cancer resections performed in a consecutive 52-month period (January 1, 2003, through April 30, 2007). SETTING: The study was performed at an 851-bed community hospital. Seventeen surgeons performed colon resections, with the number of resections varying from 1 to 154. Ten pathologists and 3 pathology assistants evaluated the specimens. PATIENTS: A total of 430 patients met the inclusion criteria and underwent surgical resection. Only patients with colon cancer were included in the study; patients with rectal cancers, in situ disease only, T4 tumors, and stage IV disease at the time of diagnosis were excluded to ensure a uniform group of patients, all undergoing resection with curative intent. MAIN OUTCOME MEASURES: Age of the patient; the surgeon, pathologist, and pathology technician; stage of disease; and year of surgery were analyzed. RESULTS: No statistical difference was found in the number of lymph nodes retrieved based on the surgeon (P = .21), pathologist (P = .11), or pathology technician (P = .26). Age of the patient, primary site of the tumor, stage, and year of surgery were all significantly associated with number of lymph nodes retrieved (P <.001). CONCLUSIONS: The origin of a low lymph node count appears multifactorial. Inadequate lymph node retrieval for colon cancer resections cannot uniformly be attributed to 1 factor, such as the surgeon. PMID- 20026829 TI - Hospital performance in caring for injured patients: does the type of injury make a difference? AB - OBJECTIVE: To determine whether quality measures based on injury-specific models provide a different perspective about relative hospital rankings compared with a single outcome measure based on all trauma patients. DESIGN: We customized the Trauma Mortality Probability Model to create separate injury-specific models for patients who sustained blunt trauma, gunshot wounds, pedestrian trauma, or motor vehicle accident trauma. SETTING: This analysis was conducted using the National Trauma Data Bank. We limited the study to hospitals with 250 or more trauma admissions per year, which coded more than 90% of patients. PATIENTS: The final data set included 54 859 patients admitted to 44 hospitals. MAIN OUTCOME MEASURES: We performed hospital-level analyses to examine the correlation between hospital risk-adjusted mortality measures based on all trauma patients vs quality measures based on injury-specific measures. RESULTS: The analysis of the intraclass correlation coefficients suggests fair-to-substantial agreement (0.39 0.68) between the hospital-adjusted odds ratios based on all patients vs odds ratios based on specific injuries. kappa Analysis demonstrated poor-to-fair agreement between hospital categorical quality measures (high, intermediate, and low quality) when hospital quality was based on outcomes for all trauma patients vs specific subgroups of patients (0.0-0.38). However, none of the hospitals classified as high quality, based on data from all trauma patients, was found to be low quality for any specific injury populations. CONCLUSION: A single composite measure based on all injured patients may not capture all the differences in hospital quality across different populations of injured patients. PMID- 20026830 TI - Endolaparoscopic approach vs conventional open surgery in the treatment of obstructing left-sided colon cancer: a randomized controlled trial. AB - OBJECTIVE: To compare self-expanding metal stents with emergency open surgery in the treatment of obstructing left-sided colon cancer. DESIGN: A randomized controlled trial. SETTING: An acute care hospital. PATIENTS: Adult patients with an obstructing tumor between the splenic flexure and rectosigmoid junction. MAIN OUTCOME MEASURES: Successful 1-stage operation, cumulative operative time, blood loss, hospital stay, pain score, and postoperative complications. RESULTS: Forty eight patients were analyzed. Twenty-four underwent endoluminal stenting followed by laparoscopic resection and 24 underwent emergency open surgery. The 2 groups were matched for age, sex, body mass index, and disease staging. Patients in the endolaparoscopic group had significantly less cumulative blood loss and lower pain, incidence of anastomotic leak, and wound infection. Significantly more patients in the endolaparoscopic group had a successful 1-stage operation performed (16 vs 9, P = .04). None of the patients in the endolaparoscopic group had a permanent stoma compared with 6 patients in the emergency open surgery group (P = .03). CONCLUSIONS: Self-expanding metal stents serve as a safe and effective bridge to subsequent laparoscopic surgery in patients with obstructing left-sided colon cancer. This endolaparoscopic approach makes a 1-stage operation more feasible, is associated with reduced incidence of stoma creation, and allows patients with malignant large-bowel obstruction to enjoy the full benefit of minimally invasive surgery. Trial Registration clinicaltrials.gov Identifier: NCT00654212. PMID- 20026831 TI - Can aviation-based team training elicit sustainable behavioral change? AB - OBJECTIVE: To quantify effects of aviation-based crew resource management training on patient safety-related behaviors and perceived personal empowerment. DESIGN: Prospective study of checklist use, error self-reporting, and a 10-point safety empowerment survey after participation in a crew resource management training intervention. SETTING: Seven hundred twenty-two-bed university hospital; 247-bed affiliated community hospital. PARTICIPANTS: There were 857 participants, the majority of whom were nurses (50%), followed by ancillary personnel (28%) and physicians (22%). MAIN OUTCOME MEASURES: Preoperative checklist use over time; number and type of entries on a Web-based incident reporting system; and measurement of degree of empowerment (1-5 scale) on a 10-point survey of safety attitudes and actions given prior to, immediately after, and a minimum of 2 months after training. RESULTS: Since 2003, 10 courses trained 857 participants in multiple disciplines. Preoperative checklist use rose (75% in 2003, 86% in 2004, 94% in 2005, 98% in 2006, and 100% in 2007). Self-initiated reports increased from 709 per quarter in 2002 to 1481 per quarter in 2008. The percentage of reports related to environment as opposed to actual events increased from 15.9% prior to training to 20.3% subsequently (P < .01). Perceived self-empowerment, creating a culture of safety, rose by an average of 0.5 point in all 10 realms immediately posttraining (mean [SD] rating, 3.0 [0.07] vs 3.5 [0.05]; P < .05). This was maintained after a minimum of 2 months. There was a trend toward a hierarchical effect with participants less comfortable confronting incompetence in a physician (mean [SD] rating, 3.1 [0.8]) than in nurses or technicians (mean [SD] rating, 3.4 [0.7] for both) (P>.05). CONCLUSIONS: Crew resource management programs can influence personal behaviors and empowerment. Effects may take years to be ingrained into the culture. PMID- 20026832 TI - Laparoscopic total gastrectomy with D2 lymph node dissection for gastric cancer. AB - OBJECTIVE: To evaluate the safety and effectiveness of laparoscopic total gastrectomy with D2 lymphadenectomy for gastric cancer. DESIGN: Review of findings from a prospectively acquired institutional database. SETTING: University hospital. PATIENTS: Fifty-five consecutive patients operated on by the same surgeon between October 1997 and March 2008. MAIN OUTCOME MEASURES: Blood loss, complication rate, and survival. RESULTS: All operations were accomplished without conversion to open laparotomy. The median operative time was 406 minutes. The median blood loss was 102 mL. A median of 46 lymph nodes were harvested. The TNM stages of the tumor were I in 17 patients (31%), II in 12 (22%), III in 16 (29%), and IV in 10 (18%). A total of 21 complications occurred in 18 patients (33%) with no postoperative mortality. At last follow-up, 44 of the 55 patients were alive without tumor recurrence and 3 with recurrence at a median follow-up of 16 months, whereas 8 had died of recurrence or another cause. CONCLUSIONS: The mortality rate of zero and acceptable morbidity of our series indicate that laparoscopic total gastrectomy with D2 lymphadenectomy is technically feasible and safe in the hands of experienced surgeons. Long-term follow-up is mandatory to validate oncologic outcome. PMID- 20026833 TI - Clinical relevance of the TLR4 11367 polymorphism in patients with major trauma. AB - OBJECTIVE: To investigate the clinical relevance of the TLR4 11367 polymorphism in patients with major trauma. DESIGN: Genetic functional and association study. SETTING: Daping Hospital and Chongqing Emergency Medical Center, Chongqing, China. PATIENTS: A total of 132 patients with major trauma were prospectively recruited. MAIN OUTCOME MEASURES: The TLR4 11367 polymorphism was genotyped using single-tube, bidirectional, allele-specific amplification method. Whole peripheral blood samples obtained within 24 hours after admission were stimulated with lipopolysaccharide and then tested for production of tumor necrosis factor alpha and interleukin 6. Sepsis morbidity rate and multiple organ dysfunction scores were assessed. RESULTS: The 11367 polymorphism was shown to be strongly associated with less capacity of peripheral leukocytes to produce tumor necrosis factor alpha and interleukin 6 in response to ex vivo lipopolysaccharide stimulation in patients with trauma at admission. Results from association study indicated that patients with trauma who carry the 11367C allele were less likely to have sepsis and multiple organ dysfunction. CONCLUSIONS: Combined with our previous in vitro functional study, the results suggest that the TLR4 11367 polymorphism might be a good predictor of who is more likely to develop complications such as sepsis or multiple organ dysfunction syndrome, depending on genotype. PMID- 20026834 TI - Preoperative weight loss before bariatric surgery. AB - HYPOTHESIS: Preoperative weight loss reduces the frequency of surgical complications in patients undergoing bariatric surgery. DESIGN: Review of records of patients undergoing open or laparoscopic gastric bypass. SETTING: A comprehensive, multidisciplinary obesity treatment center at a tertiary referral center that serves central Pennsylvania. PATIENTS: A total of 881 patients undergoing open or laparoscopic gastric bypass from May 31, 2002, through February 24, 2006. INTERVENTION: All preoperative patients completed a 6-month multidisciplinary program that encouraged a 10% preoperative weight loss. MAIN OUTCOME MEASURES: Loss of excess body weight (EBW) and total and major complication rates. RESULTS: Of the 881 patients, 592 (67.2%) lost 5% or more EBW and 423 (48.0%) lost more than 10% EBW. Patients referred for open gastric bypass (n = 466) were generally older (P < .001), had a higher body mass index (P < .001), and were more often men (P < .001) than those undergoing laparoscopic gastric bypass (n = 415). Total and major complication rates were higher in patients undergoing open gastric bypass (P < .001 and P = .03, respectively). Univariate analysis revealed that increasing preoperative weight loss is associated with reduced complication frequencies for the entire group for total complications (P =.004) and most likely for major complications (P = .06). Controlling for age, sex, baseline body mass index, and type of surgery in a multiple logistic regression model, increased preoperative weight loss was a predictor of reduced complications for any (P =.004) and major (P = .03) complications. CONCLUSION: Preoperative weight loss is associated with fewer complications after gastric bypass surgery. PMID- 20026835 TI - Factors predicting morbidity and mortality in emergency colorectal procedures in elderly patients. AB - OBJECTIVE: To identify rapidly modifiable risk factors that would improve surgical outcomes in elderly patients undergoing emergent colorectal procedures who are at high risk for morbidity and mortality. DESIGN: Retrospective review. Patients were identified on the basis of Current Procedural Terminology codes and admission through the emergency department. Medical records were reviewed and data were abstracted for comorbidities, procedural details, and in-hospital morbidity and mortality. SETTING: University tertiary referral center. PATIENTS: Two hundred ninety-two patients 65 years or older undergoing emergency colorectal procedures from January 1, 2000, through December 31, 2006. MAIN OUTCOME MEASURES: Postoperative morbidity (intensive care unit days, ventilator days, pneumonia, deep venous thrombosis, pulmonary embolus, myocardial infarction, and cerebrovascular accident) and mortality. RESULTS: The most frequent presenting diagnoses were obstructing or perforated colorectal carcinoma (30%) and perforated diverticulitis (25%). Average age at presentation was 78.1 years, and in-hospital mortality was 15%. One hundred one patients (35%) experienced a total of 195 complications. Pneumonia (25%), persistent or recurrent respiratory failure (15%), and myocardial infarction (12%) were the most frequent complications. Operative time, shock, renal insufficiency, and significant intra abdominal contamination or frank peritonitis were associated with morbidity. Age, septic shock at presentation, large estimated intraoperative blood loss, delay to operation, and development of a complication were associated with in-hospital mortality. CONCLUSIONS: Emergent colorectal procedures in the elderly are associated with significant morbidity and mortality. Minimizing the delay to definitive operative care may improve outcomes. These procedures frequently involve locally advanced colorectal cancer, emphasizing the need for improved colorectal cancer screening. PMID- 20026836 TI - Extended drainage of the pancreatic duct after pancreaticoduodenectomy. AB - HYPOTHESIS: Because of better survival following pancreaticoduodenectomy (PD), patients may develop complications due to PD and not due to malignancy per se. Exocrine insufficiency may be related to pancreatic duct obstruction or strictures attributable to duct-to-mucosa anastomosis, as in pancreaticojejunostomy. We propose a technique of managing a post-PD duct obstruction. DESIGN: Retrospective review from September 2005 to August 2008. SETTING: Methodist Dallas Medical Center, Dallas, Texas, a referral, high-volume, nonuniversity tertiary care center. PATIENTS: All patients who underwent surgery for anastomotic pancreaticojejunal stricture. MAIN OUTCOME MEASURES: Perioperative outcomes. RESULTS: All the patients were women and aged 62, 78, and 45 years. Comorbidities were documented in 2 patients. Two patients presented with severe acute abdominal pain and hyperamylasemia while 1 was asymptomatic. Two patients underwent magnetic resonance cholangiopancreatography with secretin stimulation. Endoscopic retrograde cholangiopancreatography was attempted in 1 patient. Operating time was 99 minutes, 158 minutes, and 154 minutes. Estimated blood loss was 250 mL, 400 mL, and 500 mL. A single-layer, side-to-side pancreaticogastrostomy was performed as the drainage procedure in all patients. There was no mortality associated with any of the patients within 30 days. Morbidity was seen only in 1 patient. None of the patients needed a reoperation. The mean length of hospital stay was 9 days. All patients were asymptomatic for pain. CONCLUSION: We propose a durable technique for treating pancreatic ductal strictures post-PD that appears to result in superior postoperative outcome. PMID- 20026837 TI - Prospective clinical trials of thyroidectomy with LigaSure vs conventional vessel ligation: a systematic review and meta-analysis. AB - OBJECTIVE: To evaluate the hemostatic effects and safety of thyroidectomy performed using the LigaSure vessel-sealing device (Valleylab, Boulder, Colorado) or the conventional vessel ligation. DATA SOURCES: The MEDLINE, EMBASE, Elsevier, SpringerLink, Ovid, and Cochrane Library electronic databases as well as the LigaSure manufacturer's Web site were searched for studies published between 1996 and 2008. No language restrictions were applied. STUDY SELECTION: Prospective, controlled clinical trials, both randomized and nonrandomized, comparing the hemostatic effects and safety of thyroidectomy using LigaSure and conventional vessel ligation were selected. DATA EXTRACTION: Data regarding operative parameters, duration of the operation, amount of intraoperative blood loss, length of hospital stay, and any postoperative complications were entered and analyzed using dedicated software from the Cochrane Collaboration. DATA SYNTHESIS: Four randomized and 5 nonrandomized trials that met selection criteria reported data from 927 patients, of whom 467 (50.4%) underwent LigaSure and 460 (49.6%) underwent conventional thyroidectomy. Operative duration (weighted mean difference [WMD], -11.97 minutes; 95% confidence interval [CI], -16.42 to -7.53 minutes) was significantly reduced with LigaSure thyroidectomy (P < .001). When LigaSure was used, operative time reductions of 20.32 minutes (95% CI, -33.86 to 6.79 minutes) for total thyroidectomy (P = .003) and 21.74 minutes (-38.32 to 5.16 minutes) for subtotal thyroidectomy (P = .01) were also confirmed with subgroup analysis. However, differences in the amount of intraoperative blood loss (WMD, -25.13 mL; 95% CI, -68.45 to 18.18 mL; P = .26), length of hospital stay (WMD, -0.08 days; 95% CI, -0.23 to 0.08 days; P = .31), and postoperative complication rates (odds ratio, 0.91; 95% CI, 0.61-1.04; P = .65) were not statistically significant for LigaSure vs conventional thyroidectomy. CONCLUSIONS: The LigaSure technique may provide a safe, effective, and fast alternative to conventional vessel ligation in thyroidectomy and may result in a significant reduction in operative duration. However, it may not confer any advantage over conventional thyroidectomy in terms of the amount of intraoperative blood loss, length of hospital stay, and postoperative complication rates. PMID- 20026838 TI - A unique case of recurrent metachronous volvulus of the gastrointestinal tract. AB - Colonic volvulus is an uncommon disease that predisposes patients to bowel obstruction in both the adult and pediatric population. The international literature offers few reports of synchronous or metachronous volvulus of 2 organs of the gastrointestinal tract. We describe a unique case of a patient who presented with recurrent metachronous volvulus of the sigmoid colon, cecum, and stomach. The patient underwent multiple operations for bowel obstruction, lysis of adhesions, and colon resection. The interesting intraoperative findings were a very long mesentery and peritoneal attachments of the intraabdominal gastrointestinal organs that made the stomach and colon extremely mobile and thus susceptible to volvulus. Prophylactic pexis of the cecum and the stomach during the first operation, in light of the elongated mesentery, may have prevented the subsequent episodes of volvulus. PMID- 20026839 TI - Alessandro Codivilla and the first pancreatoduodenectomy. PMID- 20026840 TI - Image of the month--quiz case. Mucinous cystadenoma of the appendix. PMID- 20026841 TI - Image of the month--quiz case. Intramesosigmoid hernia. PMID- 20026843 TI - Lichen planopilaris treated with a peroxisome proliferator-activated receptor gamma agonist. AB - Primary cicatricial alopecias (PCAs), rare disorders that lead to permanent hair loss, have been poorly understood and are difficult to treat. Lichen planopilaris (LPP)is a prototypical PCA; patients often present with sudden onset of hair loss and clinically significant symptoms of itching, burning, and pain of the scalp. Examination reveals patchy alopecia or a more diffuse thinning of the scalp with characteristic perifollicular erythema and perifollicular scale at the margins of the areas of alopecia.Treatment typically includes use of antiinflammatory medications; although symptoms may improve,hair loss is often progressive. PMID- 20026844 TI - Disparity in melanoma: a trend analysis of melanoma incidence and stage at diagnosis among whites, Hispanics, and blacks in Florida. AB - OBJECTIVE: To examine and compare the temporal trends in melanoma incidence and stage at diagnosis among whites, Hispanics, and blacks in Florida from 1990 to 2004. DESIGN: Cross-sectional and retrospective analysis. SETTING: Florida Cancer Data System. PATIENTS: Melanoma cases with known stage and race/ethnicity reported from 1990 to 2004. MAIN OUTCOME MEASURES: Age-adjusted melanoma incidence and stage at diagnosis. RESULTS: Of 41 072 cases of melanoma, 39 670 cases were reported for white non-Hispanics (WNHs), 1148 for white Hispanics (WHs), and 254 for blacks. Melanoma incidence rates increased by 3.0% per year among WNH men (P < .001), 3.6% among WNH women (P < .001), 3.4% among WH women (P = .01), and 0.9% among WH men (P = .52), while remaining relatively stable among black men and women. Both WHs and blacks had significantly more advanced melanoma at presentation: 18% of WH and 26% of black patients had either regional or distant-stage melanoma at diagnosis compared with 12% of WNH patients. The proportion of distant-stage melanoma diagnosed among WHs and blacks changed little from 1990 to 2004, compared with a steady decrease in the percentage of melanoma cases diagnosed at distant stage among WNHs (P < .001). Such differences in the time trends of the proportion of distant-stage melanoma remained after excluding in situ cases. CONCLUSIONS: The rising melanoma incidence among WNHs and WHs emphasizes the need for primary prevention. The persistence of disparity in melanoma stage at diagnosis among WHs, blacks, and WNHs warrants closer examination of secondary prevention efforts in minority groups. PMID- 20026845 TI - Factors that affect skin aging: a cohort-based survey on twins. AB - OBJECTIVE: To identify environmental factors that correlate with skin photoaging, controlling for genetic susceptibility by using a questionnaire administered to twins. DESIGN: The survey collected information about each participant's Fitzpatrick type, history of skin cancer, smoking and drinking habits, and weight from a cohort of twins. Clinicians then assigned a clinical photodamage score to each participant. SETTING: The annual Twins Days Festival in Twinsburg, Ohio. PARTICIPANTS: A voluntary cohort of twins from the general community, mostly from Ohio, Pennsylvania, and the northeastern United States. The survey was completed on a voluntary basis by sets of monozygotic (MZ) and dizygotic (DZ) twins. A total of 130 surveys taken by 65 complete twin pairs were analyzed. MAIN OUTCOME MEASURE: Skin aging was assessed using a validated photographic scale of photodamage, graded by such characteristics as wrinkling and pigmentation change. RESULTS: Photodamage scores among twins of a pair, whether MZ or DZ, were highly correlated (P = .92). Factors found to predict higher photodamage include history of skin cancer (P < .001), zygosity status (MZ vs DZ) (P = .001), weight (P = .02), and cigarette smoking (P = .046). Alcohol consumption was significantly associated with lower photodamage scores (P = .003). CONCLUSIONS: The study of twins provides a unique opportunity to control for genetic susceptibility in order to elucidate environmental influences on skin aging. The relationships found between smoking, weight, sunscreen use, skin cancer, and photodamage in these twin pairs may help to motivate the reduction of risky behaviors. PMID- 20026846 TI - Association between frequency of pruritic symptoms and perceived psychological stress: a Japanese population-based study. AB - OBJECTIVE: To evaluate the relationship between frequency of pruritic symptoms experienced over a 1-month period and psychological stress. DESIGN: Cohort study. SETTING: Population-based study in Japan. PARTICIPANTS: A total of 2224 participants at least 18 years old and without psychiatric disorders participated in the Japan Health Diary Study (October 2003), a cohort study comprising a representative sample in Japan. MAIN OUTCOME MEASURES: Frequency of pruritic symptoms assessed by self-reported health diaries over the 1-month period and subsequent psychological stress measured using the Japanese version of the Perceived Stress Scale. RESULTS: The 2224 participants had a mean age of 44.6 years, 1212 (54.5%) were women, and 70 (3.1%) presented with pruritic symptoms. Multivariable analysis showed that patients with pruritic symptoms had significantly higher psychological stress than those without pruritic symptoms (beta coefficient, 2.33; 95% confidence interval [CI], 0.53-4.14; P = .01). Furthermore, a linear trend was observed between increased psychological stress and increased severity of pruritic symptoms, with beta coefficients for the first, second, and third tertiles for symptoms of 0.81 (95% CI, -1.97 to 3.59), 1.77 (95% CI, -0.82 to 4.37), and 4.86 (95% CI, 1.29 to 8.43), respectively (P value for trend, .004). CONCLUSION: Our results suggest that frequency of pruritic symptoms is associated with psychological stress in the general population. PMID- 20026848 TI - Applicability and prognostic value of the new TNM classification system in 135 patients with primary cutaneous anaplastic large cell lymphoma. AB - OBJECTIVES: To test the applicability and prognostic value of the new TNM classification system for primary cutaneous lymphomas other than mycosis fungoides and Sezary syndrome in patients with primary cutaneous anaplastic large cell lymphoma (C-ALCL) and to evaluate the prognostic significance of other clinical variables, in particular the site of presentation. DESIGN: Retrospective cohort analysis. SETTING: Dutch Cutaneous Lymphoma Group database. Patients One hundred thirty-five patients with C-ALCL. MAIN OUTCOME MEASURES: Clinical variables, including T category and site of presentation. RESULTS: Eighty patients (59.3%) presented with T1 disease, 37 (27.4%) with T2 disease, and 18 (13.3%) with T3 disease. Median follow-up was 56 months (range, 11-288 months). Five-year disease-specific survival (DSS) was 93% for T1 disease, 93% for T2 disease, and 77% for T3 disease (P = .19). Patients with skin lesions on a leg had reduced 5-year DSS compared with lesions on other sites (82% for leg vs 95% for head and neck, 96% for trunk, and 95% for arm; P = .23). Patients with leg involvement (n = 32) had significantly worse 5-year DSS than did patients without leg involvement (n = 103; 76% vs 96%; P = .03 after adjustment for T category). CONCLUSIONS: The new TNM system can be applied well to patients with C-ALCL and may provide prognostic information, in particular when combined with site of presentation. Patients with T2 or T3 disease with skin lesions on the leg may have reduced survival and require close surveillance during follow-up. PMID- 20026847 TI - Incidence of and risk factors for skin cancer after heart transplant. AB - OBJECTIVE: To examine the incidence, tumor burden, and risk factors for nonmelanoma and other skin cancer types in this heart transplant cohort. DESIGN: Retrospective review of patient medical records. SETTING: Tertiary care center. Patients All heart transplant recipients at Mayo Clinic from 1988 to 2006. MAIN OUTCOME MEASURES: Cumulative incidence of skin cancer and tumor burden, with Cox proportional hazards regression models used to evaluate risk factors for posttransplant primary and secondary nonmelanoma skin cancer. RESULTS: In total, 312 heart transplant patients had 1395 new skin cancers in 2097 person-years (mean, 0.43 per year per patient) with a range of 0 to 306 for squamous cell carcinoma (SCC) and 0 to 17 for basal cell carcinoma (BCC). The cumulative incidence rates of any skin cancer were 20.4%, 37.5%, and 46.4% at 5, 10, and 15 years after heart transplant, respectively. Cumulative incidence of SCC after the first BCC was 98.1% within 7 years. Multivariate analysis showed that posttransplant nonskin cancer, increased age, and heart failure etiologic factors other than idiopathic disease were associated with increased risk of SCC. Posttransplant herpes simplex viral infection, increased age, and use of mycophenolate mofetil for immunosuppression were associated with increased risk of BCC. CONCLUSIONS: With prolonged survival, many heart transplant patients have numerous skin cancers. Vigilant sun protection practices, skin cancer education, and regular skin examination are appropriate interventions in these high-risk patients. PMID- 20026850 TI - Follicular red dots: a novel dermoscopic pattern observed in scalp discoid lupus erythematosus. AB - BACKGROUND: Scalp dermoscopy plays an important role in the diagnosis of hair and scalp disorders, and specific dermoscopic patterns have recently been associated with several disorders causing noncicatricial alopecia, such as androgenetic alopecia, alopecia areata, trichotillomania, and tinea capitis. OBSERVATIONS: We describe the morphologic and pathologic features of a new dermoscopic pattern, referred to as "follicular red dots," that was found in scalp lesions of 5 patients with active discoid lupus erythematosus (DLE). Follicular red dots appear as erythematous polycyclic, concentric structures, with a diameter ranging from 0.16 to 0.47 mm, regularly distributed in and around the follicular ostia. In the pathologic findings, red dots correspond to widened infundibula plugged by keratin and surrounded by dilated vessels and extravasated erythrocytes. Retrospective blinded evaluation of the dermoscopic images of 155 patients with cicatricial alopecia suggests that follicular red dots are a specific feature of DLE because the pattern was not identified in the images of cicatricial alopecia resulting from other diseases. CONCLUSIONS: The follicular red dot pattern is a specific feature of scalp lesions of active lupus erythematosus of the scalp. Recognition of this distinctive dermoscopic pattern may help the clinician to differentiate DLE from other diseases causing cicatricial alopecia. PMID- 20026851 TI - A rectangular dermatosis of the left back. AB - BACKGROUND: Cardioversion and defibrillation have become widely used techniques aimed at restoring normal sinus rhythm in patients with cardiac arrhythmias. Following the procedure, cutaneous lesions are often seen at the site of the electrodes, but little has been reported regarding the evolution of such lesions over time. OBSERVATIONS: Two patients presented with unusual, well-defined rectangular eruptions on the left back, and both reported a history of having undergone electrical cardioversion or defibrillation several years previously. The histologic characteristics of each lesion were distinct, and the management was symptomatic, with most of the relief coming from the recognition that the eruption was actually a self-limited manifestation of cardioversion and defibrillation. CONCLUSIONS: The clinical cases and corresponding histologic findings represent possible long-term sequelae of electrical cardioversion or defibrillation. They are presented in order to enhance the diagnostic acumen of dermatologists and to avoid potential misdiagnosis. PMID- 20026852 TI - Psychological responses and coping strategies among patients with malignant melanoma: a systematic review of the literature. AB - OBJECTIVE: To conduct a systematic review of the literature to identify the prevalence of, and demonstrated risk factors for, psychological distress among individuals affected by, or at high risk of developing, melanoma. For a substantial subset of patients, the diagnosis and/or treatment of cutaneous malignant melanoma may cause significant psychological distress. DATA SOURCES: Using the MEDLINE, PsycINFO, and CINAHL databases, published studies (1988 to March 2008) of individuals affected by melanoma were included if they examined the demographic, clinical, psychological, and/or social correlates of emotional distress. STUDY SELECTION: Searches were restricted to publications in English and were supplemented by citation lists in retrieved articles and contact with researchers. DATA EXTRACTION: A total of 356 articles were critically appraised by 2 reviewers to assess eligibility and clinical evidence level. A total of 44 studies met the inclusion criteria. DATA SYNTHESIS: Approximately 30% of patients with melanoma reported clinically relevant levels of psychological distress, as measured by a range of validated scales, with symptoms of anxiety more prevalent than depression. A number of empirically demonstrated risk factors for distress were identified, including female sex, younger age, lower education, visibility of affected body site, lack of social support, and negative appraisal of melanoma. CONCLUSIONS: Routine psychological screening of patients with melanoma is widely recommended as standard practice; however, standard screening measures may have limited sensitivity and specificity as demonstrated by the wide range of results reported in this review. Development of a brief screening tool that incorporates empirically supported risk factors is recommended to improve the timely identification and support of those patients most susceptible to adverse psychological outcomes. PMID- 20026853 TI - Mohs micrographic surgery for Basal cell carcinoma of the face. PMID- 20026854 TI - Topical imiquimod or fluorouracil therapy for basal and squamous cell carcinoma: a systematic review. AB - OBJECTIVES: To conduct a systematic review to determine clearance rates and adverse effects of topical imiquimod or fluorouracil therapy in the treatment of nonmelanoma skin cancers such as basal (BCC) and squamous cell carcinoma (SCC), and to develop recommendations for the use of topical imiquimod or fluorouracil to treat BCC and SCC. DATA SOURCES: MEDLINE, CANCERLIT, and Cochrane databases. STUDY SELECTION: Prospective, retrospective, and case studies in English containing a minimum of 4 subjects and a 6-month follow-up or posttreatment histologic evaluation. DATA EXTRACTION: We calculated the rate of clearance and adverse effects for BCC subtypes and invasive and in situ SCC treated with topical imiquimod or fluorouracil. DATA SYNTHESIS: Clearance rates varied by drug regimen, and most of the studies lacked long-term follow-up. Imiquimod use produced the following clearance rates: 43% to 100% for superficial BCC, 42% to 100% for nodular BCC, 56% to 63% for infiltrative BCC, 73% to 88% for SCC in situ, and 71% for invasive SCC. Fluorouracil use produced the following clearance rates: 90% for superficial BCC and 27% to 85% for SCC in situ. Up to 100% and 97% of patients applying imiquimod and fluorouracil, respectively, experienced at least 1 adverse event. Adverse event intensity ranged from mild to severe; erythema, pruritus, and pain were common. CONCLUSIONS: Evidence supports the use of topical imiquimod as monotherapy for superficial BCC and topical fluorouracil as monotherapy for superficial BCC and SCC in situ. Based on the available evidence, the strength of any recommendations for the use of these 2 agents in the primary treatment of these tumors is weak. We recommend that their use be limited to patients with small tumors in low-risk locations who will not or cannot undergo treatment with better-established therapies for which long-term clearance rates have been determined. Long-term clinical follow-up is essential for patients treated with topical imiquimod or fluorouracil. Limitations of therapy include high rates of adverse effects, lower clearance rates than other treatment modalities, dependence on patient adherence to treatment, and higher costs than other therapies. PMID- 20026855 TI - Psychoneuro-oncology: its time has arrived. PMID- 20026856 TI - A conceptual framework for advancing melanoma health disparities research. PMID- 20026857 TI - Multiple comedonelike lesions encircling the anal orifice--quiz case. PMID- 20026858 TI - A cherry-red umbilical papule in an infant--quiz case. PMID- 20026859 TI - Sharply demarcated, erythematous plaques on an amputation stump--quiz case. PMID- 20026860 TI - Widespread crusted lesions in a patient with HIV--quiz case. PMID- 20026861 TI - The fascial plication suture: an adjunct to layered wound closure. PMID- 20026862 TI - Lack of lower extremity hair not a predictor for peripheral arterial disease. PMID- 20026863 TI - Association between thin melanomas and atypical nevi in middle-aged and older men possibly attributable to heightened patient awareness. PMID- 20026864 TI - A case of lichen sclerosus of the scalp associated with autoantibodies to extracellular matrix protein 1. PMID- 20026865 TI - Septic facial vein thrombosis due to Panton-Valentine leukocidin-positive Staphylococcus aureus. PMID- 20026866 TI - Onychomatricoma: clinical and sonographic findings. PMID- 20026867 TI - Dermoscopy of extragenital lichen sclerosus. PMID- 20026868 TI - Angiotensin-converting enzyme inhibitors versus angiotensin receptor blockers for diabetic nephropathy: a retrospective comparison. AB - INTRODUCTION: There are no adequate head-to-head comparisons of angiotensin converting enzyme (ACE) inhibitors and angiotensin receptor blockers (ARB) in type 2 diabetic patients in spite of some interesting attempts. Furthermore, there are no adequate studies about the effects of ACE inhibitors in type 2 diabetic patients, who are the great majority of diabetic individuals. This study has retrospectively compared the effects of ACE inhibitors and ARBs used to treat diabetic nephropathy in a group of type 2 diabetic subjects. DESIGN AND METHODS: Patients (n=154) were treated with ACE inhibitors (mean age 59.5+/-13.3 years, 52.6% were male). Eighty-five patients had been treated with ARBs from 1999 until now (mean age 62.6+/-10.9 years, 56.0% were male, differences not significant). Kaplan-Meier survival analysis was used to calculate survival before reaching end stage renal disease (ESRD) (glomerular filtration < 15 ml/min, stage V of renal disease as defined by KDOQI clinical guidelines) or starting renal replacement therapy. Only patients treated for more than six months were included in the survival analysis. Comparison of survival was made at three, five and seven years after starting treatment. RESULTS: Pre-ESRD survival was 91.9% at three years, 81.6% at five years and 61.9% at seven years of follow-up for patients treated with ACE inhibitors. For patients treated with ARBs, pre-ESRD survival was 95.3% at three years, 82.1% at five years and 78.2% at seven years of follow-up (p=0.02, log-rank test). At 36 months, the comparative odds ratio for having started renal replacement therapy or reaching end-stage renal failure was 0.246 (95% confidence interval 0.114-0.531, p<0.001 for chi-square and likelihood ratio tests). The risk for the ARB cohort was 0.682 (95% confidence interval 0.578 0.804), meanwhile for ACE inhibitor patients it was 2.768 (95% confidence interval 1.481-5.172). CONCLUSIONS: The effects of ACE inhibitors and ARBs seem to be different, favouring the use of ARBs. These results may have been influenced by the different circumstances when each kind of drug was indicated, since ARBs were used with the specific recommendations for control of blood pressure in diabetic patients. An earlier referral of these patients may also have had some effect on these results. The need for a well-designed prospective study on type 2 diabetic patients with heavy proteinuria is warranted. PMID- 20026869 TI - Cardioprotective effects of benazepril, an angiotensin-converting enzyme inhibitor, in an ischaemia-reperfusion model of myocardial infarction in rats. AB - INTRODUCTION: The present study evaluated the effects of benazepril, an angiotensin-converting enzyme inhibitor on haemodynamic, biochemical, and immunohistochemical (Bax and Bcl-2 protein) indices in ischaemia and reperfusion (IR) injury. MATERIALS AND METHODS: Male Wistar albino rats were divided into three groups and were orally administered saline once daily (IR-sham and IR control) or benazepril (30 mg/kg/day; IR-benazepril) for 14 days. On the 15(th) day, in the IR-control and IR-benazepril groups, rats were subjected to left anterior descending coronary artery occlusion for 45 minutes followed by a one hour reperfusion. Haemodynamic parameters were recorded and rats were sacrificed; hearts were isolated for biochemical estimation and immunohistochemistry. RESULTS: In the IR-control group, significant ventricular dysfunctions (p<0.05 vs. IR-sham group) were observed along with enhanced expression of pro-apoptotic protein Bax. A decline in lactate dehydrogenase activity and increased content of thiobarbituric acid reactive substances, a marker of lipid peroxidation, were observed. Benazepril pretreatment significantly improved mean arterial pressure (p<0.01), reduced left ventricular end-diastolic pressure (p<0.05), and improved both inotropic and lusitropic function of the heart (+LVdP/dt and - LVdP/dt) (p<0.05; p<0.01) as compared to IR-control. Furthermore, benazepril treatment significantly decreased the level of thiobarbituric acid reactive substances and restored the activity of lactate dehydrogenase towards normal value (p<0.05 vs. IR-control). CONCLUSION: This study demonstrates that benazepril upregulated Bcl 2 protein and decreased Bax protein expression, thus exhibiting anti-apoptotic effects. These beneficial effects of benazepril will have an important implication in the therapeutic use of benazepril in ischaemic heart disease. PMID- 20026870 TI - Angiotensin-converting enzyme insertion/deletion and angiotensin type 1 receptor A1166C polymorphisms as genetic risk factors in benign prostatic hyperplasia and prostate cancer. AB - INTRODUCTION: Prostate cancer is one of the most common malignant neoplasias in developed countries. In 2003, 6,536 new cases and 4,602 related deaths were reported in Mexico. The renin-angiotensin system has been shown to play a role in prostate cancer pathology. Two previous studies investigated the association of prostate cancer with the insertion/deletion (I/D) polymorphism in the angiotensin converting enzyme (ACE) gene; both studies reported an association between prostate cancer and the DD genotype. The present study was aimed at searching for an association of prostate cancer and benign prostatic hyperplasia with the I/D polymorphism in the ACE gene and the A1166C polymorphism in the angiotensin type 1 receptor (AGT1R) gene and at comparing allele frequencies between both groups and the general population. MATERIALS AND METHODS: DNA was extracted from 20 samples from individuals with a prostate cancer diagnosis and from 20 samples from individuals with a benign prostatic hyperplasia diagnosis. Genotyping was performed by PCR-RFLP analysis. Polymorphism frequency results obtained for the test groups were compared with the frequencies in 66 individuals from the general population, which were previously obtained at the same molecular medicine laboratory in the context of other studies. RESULTS: The comparative analysis of the three groups revealed significant differences for allele frequencies in the two genes in patients groups (prostate cancer and benign prostatic hyperplasia) versus the general population. The D allele in the ACE gene was closely associated with a significant higher risk of developing both benign prostatic hyperplasia (odds ratio [OR]=21.87; 95% confidence interval [CI]=2.314-206.479) or prostate cancer (OR=31.66; 95% CI=0.091-1.272), and the AGT1R A1166 allele in the homozygote state was identified as a risk genotype for benign prostatic hyperplasia (OR=56.07). CONCLUSIONS: Genotypes in ACE and AGT1R polymorphisms could be considered as genetic risk markers for benign prostatic hyperplasia or prostate cancer. PMID- 20026871 TI - Effect of atomoxetine on executive function impairments in adults with ADHD. AB - OBJECTIVE: To assess the effect of atomoxetine on ADHD-related executive functions over a 6-month period using the Brown Attention-Deficit Disorder Scale (BADDS) for Adults, a normed, 40-item, self-report scale in a randomized, double blind, placebo-controlled clinical trial. METHOD: In a randomized, double-blind clinical trial, adults with ADHD received either atomoxetine 25 to 100 mg/day or placebo for 6 months. Patients completed the BADDS to report their current daily functioning in 5 clusters of ADHD-related impairments of executive functioning: (1) Organizing and Activating to Work; (2) Focusing for Tasks; (3) Regulating Alertness and Effort; (4) Modulating Emotions; and (5) Utilizing Working Memory. RESULTS: Mean scores were significantly more improved in the atomoxetine group compared to the placebo group: total score, -27.0 versus -19.0 (p < .001); all 5 cluster scores, p < .01. CONCLUSIONS: Once-daily atomoxetine can improve executive function impairments in adults with ADHD as assessed by the BADDS. PMID- 20026872 TI - Incidental encoding strategies did not improve contextual memory in Parkinson's disease patients. AB - BACKGROUND: This study investigated the performance of patients with idiopathic Parkinson's disease (PD) without dementia for incidental recognition memory and the effect of encoding strategies on contextual memory. METHODS: The authors studied 21 patients with PD (ages 60-85, 12 women; Hoehn and Yahr I-III, Activities of Daily Living 70%-100%) and 22 healthy controls (ages 60-84, 18 women). Participants completed the vocabulary subtest of the Wechsler Adult Intelligence Scale and the Wisconsin Card Sorting Test (WCST). To assess the incidental recognition memory for item (object) and context (location of the object), participants of each group were assigned to 1 of 2 encoding conditions: ( a) an incidental associative instruction to bind the object to its location or (b) a nonassociative, nonspecific instruction. RESULTS: PD patients showed performance comparable to the control group's on the vocabulary subtest and WCST. In contrast to controls, PD patients were unable to take advantage of the associative encoding instruction, which also had a deleterious effect on item recognition. CONCLUSION: This sample of participants with PD showed diminished item and context recognition memory and an impaired ability to use incidental memory encoding strategy, suggesting a compromised cognitive reserve. The fact that these alterations occurred in early stages of PD, and prior to more general cognitive alterations such as executive dysfunction, should be considered in the management of patients by using specific cognitive rehabilitation interventions. PMID- 20026875 TI - Maternal plasma DNA analysis with massively parallel sequencing by ligation for noninvasive prenatal diagnosis of trisomy 21. AB - BACKGROUND: Noninvasive prenatal diagnosis of trisomy 21 (T21) has recently been shown to be achievable by massively parallel sequencing of maternal plasma on a sequencing-by-synthesis platform. The quantification of several other human chromosomes, including chromosomes 18 and 13, has been shown to be less precise, however, with quantitative biases related to the chromosomal GC content. METHODS: Maternal plasma DNA from 10 euploid and 5 T21 pregnancies was sequenced with a sequencing-by-ligation approach. We calculated the genomic representations (GRs) of sequenced reads from each chromosome and their associated measurement CVs and compared the GRs of chromosome 21 (chr21) for the euploid and T21 pregnancies. RESULTS: We obtained a median of 12 x 10(6) unique reads (21% of the total reads) per sample. The GRs deviated from those expected for some chromosomes but in a manner different from that previously reported for the sequencing-by-synthesis approach. Measurements of the GRs for chromosomes 18 and 13 were less precise than for chr21. z Scores of the GR of chr21 were increased in the T21 pregnancies, compared with the euploid pregnancies. CONCLUSIONS: Massively parallel sequencing-by-ligation of maternal plasma DNA was effective in identifying T21 fetuses noninvasively. The quantitative biases observed among the GRs of certain chromosomes were more likely based on analytical factors than biological factors. Further research is needed to enhance the precision for measuring for the representations of chromosomes 18 and 13. PMID- 20026874 TI - The lysyl oxidases LOX and LOXL2 are necessary and sufficient to repress E cadherin in hypoxia: insights into cellular transformation processes mediated by HIF-1. AB - Hypoxia has been shown to promote tumor metastasis and lead to therapy resistance. Recent work has demonstrated that hypoxia represses E-cadherin expression, a hallmark of epithelial to mesenchymal transition, which is believed to amplify tumor aggressiveness. The molecular mechanism of E-cadherin repression is unknown, yet lysyl oxidases have been implicated to be involved. Gene expression of lysyl oxidase (LOX) and the related LOX-like 2 (LOXL2) is strongly induced by hypoxia. In addition to the previously demonstrated LOX, we characterize LOXL2 as a direct transcriptional target of HIF-1. We demonstrate that activation of lysyl oxidases is required and sufficient for hypoxic repression of E-cadherin, which mediates cellular transformation and takes effect in cellular invasion assays. Our data support a molecular pathway from hypoxia to cellular transformation. It includes up-regulation of HIF and subsequent transcriptional induction of LOX and LOXL2, which repress E-cadherin and induce epithelial to mesenchymal transition. Lysyl oxidases could be an attractive molecular target for cancers of epithelial origin, in particular because they are partly extracellular. PMID- 20026876 TI - The simplicity of completion time distributions for common complex biochemical processes. AB - Biochemical processes typically involve huge numbers of individual reversible steps, each with its own dynamical rate constants. For example, kinetic proofreading processes rely upon numerous sequential reactions in order to guarantee the precise construction of specific macromolecules. In this work, we study the transient properties of such systems and fully characterize their first passage (completion) time distributions. In particular, we provide explicit expressions for the mean and the variance of the completion time for a kinetic proofreading process and computational analyses for more complicated biochemical systems. We find that, for a wide range of parameters, as the system size grows, the completion time behavior simplifies: it becomes either deterministic or exponentially distributed, with a very narrow transition between the two regimes. In both regimes, the dynamical complexity of the full system is trivial compared to its apparent structural complexity. Similar simplicity is likely to arise in the dynamics of many complex multistep biochemical processes. In particular, these findings suggest not only that one may not be able to understand individual elementary reactions from macroscopic observations, but also that such an understanding may be unnecessary. PMID- 20026877 TI - Physico-chemical modelling of target depletion during hybridization on oligonulceotide microarrays. AB - The effect of target molecule depletion from the supernatant solution is incorporated into a physico-chemical model of hybridization on oligonucleotide microarrays. Two possible regimes are identified: local depletion, in which depletion by a given probe feature only affects that particular probe, and global depletion, in which all features responding to a given target species are affected. Examples are given of two existing spike-in data sets experiencing measurable effects of target depletion. The first of these, from an experiment by Suzuki et al using custom built arrays with a broad range of probe lengths and mismatch positions, is verified to exhibit local and not global depletion. The second data set, the well-known Affymetrix HGU133a latin square experiment, is shown to be very well explained by a global depletion model. It is shown that microarray calibrations relying on Langmuir isotherm models which ignore depletion effects will significantly underestimate specific target concentrations. It is also shown that a combined analysis of perfect match and mismatch probe signals in terms of a simple graphical summary, namely the hook curve method, can discriminate between cases of local and global depletion. PMID- 20026878 TI - Attenuation of transcriptional bursting in mRNA transport. AB - Because of the stochastic nature of biochemical processes, the copy number of any given type of molecule inside a living cell often exhibits large temporal fluctuations. Here, we develop analytic methods to investigate how the noise arising from a bursting input is reshaped by a transport reaction which is either linear or of the Michaelis-Menten type. A slow transport rate smoothes out fluctuations at the output end and minimizes the impact of bursting on the downstream cellular activities. In the context of gene expression in eukaryotic cells, our results indicate that transcriptional bursting can be substantially attenuated by the transport of mRNA from nucleus to cytoplasm. Saturation of the transport mediators or nuclear pores contributes further to the noise reduction. We suggest that the mRNA transport should be taken into account in the interpretation of relevant experimental data on transcriptional bursting. PMID- 20026880 TI - Reliability of thickness measurements of the dorsal muscles of the upper cervical spine: an ultrasonographic study. AB - STUDY DESIGN: Clinical measurement, reliability. OBJECTIVES: To examine the intraday intrarater reliability of measuring thickness of the upper dorsal neck muscles at rest, as well as at 50% of maximum voluntary isometric contraction (MVIC), for upper cervical extension. BACKGROUND: Methodology for measuring the thickness of the lower dorsal neck muscles, including semispinalis capitis and multifidus muscles, during contraction using ultrasonography has been established. Thickness measurements for the upper dorsal neck muscles have not been documented. METHODS: Ten subjects (21 to 30 years of age) without neck pain and headache were recruited. Their upper dorsal neck muscles were measured both at rest and during 50% MVIC for upper cervical extension in sitting position using rehabilitative ultrasound imaging (RUSI). Muscles measured included the rectus capitis posterior major, oblique capitis superior, semispinalis capitis, and splenius capitis. All measurements were repeated after 10 minutes of rest, on the same day, by the same rater. Descriptive statistics were supplemented by calculations of intraclass correlation coefficient (ICC3,1), standard error of measurement (SEM), within-subject coefficient of variation (CVw), and minimal detectable change (MDC). RESULTS: ICC3,1 results ranged from 0.87 to 0.99 for thickness measurements made at rest and from 0.90 to 0.98 for thickness measurements made with a 50% MVIC. The SEMs for thickness measurements at rest and at 50% MVIC ranged from 0.11 to 0.46 mm and 0.23 to 0.52 mm, while the CVws ranged from 3.5% to 6.1% and 3.7% to 6.4%, and MDC95 ranged from 0.35 to 1.46 mm and 0.73 to 1.65 mm, respectively. The thickness of all upper dorsal neck muscles measured during a 50% MVIC was greater than when measured at rest (P<.029). CONCLUSION: Thickness measurements of the upper dorsal neck muscles using RUSI were reliable both at rest and during a 50% effort isometric contraction. PMID- 20026881 TI - Conservative management of a young adult with hip arthrosis. AB - STUDY DESIGN: Case report. BACKGROUND: Clinical practice guidelines regarding the conservative management of degenerative hip conditions in older adults routinely incorporate therapeutic exercise and manual therapy. However, the application of these recommendations to young, active adults is less clear. The purpose of this case report is to describe the management of a young adult with advanced hip arthrosis using a multifaceted rehabilitation program. CASE DESCRIPTION: A 28 year-old female with severe left hip degeneration, as identified with diagnostic imaging, was referred to physical therapy. Reduced hip range of motion and strength, sacroiliac joint asymmetries, and a modified Harris Hip Score of 76 were observed. She was seen for 12 visits over a 3-month period and treated with an individualized program including manual therapy, therapeutic exercise, and neuromuscular re-education. OUTCOME: Substantial improvements were noted in pain, hip range of motion, and strength and function (modified Harris Hip Score of 97). In addition, she discontinued the use of anti-inflammatory medications and returned to her prior level of activity. Improvements were maintained at a 3 month follow-up, with symptom recurrence managed using a self-mobilization technique to the left hip and massage to the left iliopsoas. DISCUSSION: Degenerative hip conditions are common among older adults but are relatively rare in the younger population. Although it is likely that this patient will experience a return of her symptoms and functional limitations as her hip disease progresses, the immediate improvements may delay the need for eventual surgical management. These outcomes suggest that physical therapy management should be considered in those with an early onset of degenerative hip disease and are consistent with results previously reported in the older population. LEVEL OF EVIDENCE: Therapy, level 4. PMID- 20026882 TI - Effect of medial arch support on displacement of the myotendinous junction of the gastrocnemius during standing wall stretching. AB - STUDY DESIGN: Controlled laboratory study. OBJECTIVES: To examine the effects of standing wall stretching with and without medial arch support (WMAS versus WOMAS) on the displacement of the myotendinous junction (DMTJ) of the medial gastrocnemius, rearfoot angle, and navicular height in subjects with neutral foot alignment and pes planus. BACKGROUND: Standing wall stretching is often prescribed to increase ankle dorsiflexion range of motion for sports fitness and rehabilitation. However, the effect of standing wall stretching WMAS on DMTJ is unknown. METHODS: Fifteen subjects with neutral foot alignment and 15 subjects with pes planus performed standing wall stretching under WMAS and WOMAS conditions. Measurements of DMTJ and rearfoot position were performed using ultrasonography and video imaging. Navicular height was measured using a ruler. Dependent variables were examined with a 2-way mixed-design analysis of variance. The 2 factors were foot type (neutral foot versus pes planus) and stretching condition (WMAS versus WOMAS). RESULTS: There were significant interactions of medial arch support by foot type for DMTJ, rearfoot angle, and navicular drop (P<.001). A post hoc paired t test showed that standing wall stretching in the WMAS condition significantly increased the DMTJ, compared to stretching in the WOMAS condition, in subjects with neutral foot (mean +/- SD, 9.6 +/- 1.6 versus 10.5 +/- 1.6 mm; difference, 0.9 mm; 99% CI: 0.4-1.4 mm) and in those with pes planus (10.0 +/- 1.8 versus 12.7 +/- 2.0 mm; difference, 2.7 mm; 99% CI: 1.9-3.5 mm) (P<.001). When comparing WOMAS and WMAS, the difference in DMTJ (1.8 mm; 99% CI: 0.9-2.7 mm) was significantly greater in subjects with pes planus than in those with neutral foot (P<.001). CONCLUSION: Standing wall stretching with medial arch support maintained subtalar joint neutral position and increased the length of the gastrocnemius in subjects with pes planus. When prescribing standing wall stretching, clinicians need to emphasize the use of medial arch support to effectively stretch the gastrocnemius in subjects with pes planus. PMID- 20026883 TI - Efficacy of an ankle brace with a subtalar locking system in inversion control in dynamic movements. AB - STUDY DESIGN: Controlled laboratory study. OBJECTIVES: To examine effectiveness of an ankle brace with a subtalar locking system in restricting ankle inversion during passive and dynamic movements. BACKGROUND: Semirigid ankle braces are considered more effective in restricting ankle inversion than other types of brace, but a semirigid brace with a subtalar locking system may be even more effective. METHODS: Nineteen healthy subjects with no history of major lower extremity injuries were included in the study. Participants performed 5 trials of an ankle inversion drop test and a lateral-cutting movement without wearing a brace and while wearing either the Element (with the subtalar locking system), a Functional ankle brace, or an ASO ankle brace. A 2-way repeated-measures analysis of variance (ANOVA) was used to assess brace differences (P?.05). RESULTS: All 3 braces significantly reduced total passive ankle frontal plane range of motion (ROM), with the Element ankle brace being the most effective. For the inversion drop the results showed significant reductions in peak ankle inversion angle and inversion ROM for all 3 braces compared to the no brace condition; and the peak inversion velocity was also reduced for the Element brace and the Functional brace. In the lateral-cutting movement, a small but significant reduction of the peak inversion angle in early foot contact and the peak eversion velocity at push off were seen when wearing the Element and the Functional ankle braces compared to the no brace condition. Peak vertical ground reaction force was reduced for the Element brace compared to the ASO brace and the no brace conditions. CONCLUSIONS: These results suggest that the tested ankle braces, especially the Element brace, provided effective restriction of ankle inversion during both passive and dynamic movements. PMID- 20026884 TI - Lunate fracture in an amateur soccer player. PMID- 20026888 TI - [A case of eosinophilic abscess mistaken for metastasis due to FDG uptake in PET CT]. PMID- 20026889 TI - [Chemotherapy for colorectal cancer]. AB - Recent advances in chemotherapy lead to improved survival outcomes in patients with colorectal cancer. The 5-fluorouracil (5-FU) is still one of the important chemotherapeutic agents since 1950s, but the introduction of newer cytotoxic agents, irinotecan and oxaliplatin, or targeted agents, bevacizumab and cetuximab, have changed treatment strategies for these patients. A deliberate choice should be made for adjuvant chemotherapy, because it has became complicated more than ever before. Oxaliplatin plus 5-FU seemed to be superior in terms of disease-free and overall survival than 5-FU alone after curative surgery for colon cancers. However not all of these patients seemed to receive benefit from this intensive adjuvant treatment, and some limitations are present according to the postoperative stage, tumor biology and clinical characteristics. For metastatic disease, there is no doubt that more complicated strategies are present because we have more abundant chemotherapeutic agents available for metastatic setting compared to adjuvant setting. Recently, targeted agents, such as bevacizumab or cetuximab, also took an important place in the treatment of metastatic colorectal cancer, and many efforts are also made to find the biomarkers for predicting treatment responses to these targeted agents. In this review, we intended to sort up the standard strategies of chemotherapy for patients with colorectal cancer according to the latest pivotal publications. PMID- 20026890 TI - [Clinical practice patterns of gastroenterologists for initial and maintenance therapy in gastroesophageal reflux disease: a nationwide online survey in Korea]. AB - BACKGROUND/AIMS: Gastroesophageal reflux disease (GERD) is a chronic condition and accompanied by frequent relapses. We aimed to evaluate the clinical practice patterns of gastroenterologists for initial and maintenance therapy of GERD in Korea. METHODS: We administered a nationwide, multi-center, and questionnaire based online survey between December 2007 and January 2008. The questionnaire consisted of 15 questions about prescribing patterns of initial and maintenance therapy for GERD. RESULTS: A total of 371 gastroenterologists participated in the survey with the response rate of 77%. For mild cases of GERD, the most common choice of initial therapy was full dose proton-pump inhibitors (PPIs) (59%), followed by half dose PPIs (20%), and H2 receptor antagonists (4%). For severe cases, full dose PPIs were prescribed in 99%. Almost all gastroenterologists agreed to the need for maintenance therapy. For both mild (95%) and severe (99%) cases of GERD, gastroenterologists preferred the use of PPI-based maintenance regimen. The preferred maintenance strategy for GERD was continuous therapy in erosive esophagitis (67%), and on-demand therapy in non-erosive reflux disease (68%). The overall duration of the therapy (initial+maintenance) was 7.7+/-5.1 wk in mild cases and 15.0+/-9.4 wk in severe cases. The duration of maintenance therapy was affected by symptom severity, followed by symptom frequency and endoscopic finding. CONCLUSIONS: In this study, a majority of the gastroenterologists is aware of importance on PPI-based pharmacological treatment for GERD. Further studies are needed to clarify the appropriate strategy and duration of maintenance therapy. PMID- 20026891 TI - [Colonoscopic perforation; a 10-year experience in single general hospital]. AB - BACKGROUND/AIMS: Colonoscopy is the principal method for diagnosis, treatment, and follow up of colorectal disease. The study aimed to assess the incidence, clinical features, and management of colonoscopic perforations at a local general hospital. METHODS: A retrospective review of patient record was performed for all patients with iatrogenic colonic perforation after sigmoidoscopy and colonoscopy between 1997 and 2007. RESULTS: In the 10-year period, 16,388 colonoscopic and sigmoidscopic procedure were performed. All 10 cases of procedure related colonic perforation were developed. Perforation occurred in 9 cases during therapeutic procedure; 5 cases due to polypectomy and 4 cases due to endoscopic submucosal dissection. Perforation occurred in one case during diagnostic procedure. CONCLUSIONS: Therapeutic procedure is a clear risk factor of colonic perforation. When colonic perforation occurs, we should be able to make early diagnosis. Early diagnosis can lead to a good treatment and can produce good prognosis with short hospital days. PMID- 20026892 TI - [Association of Toll-like receptor gene with Crohn's disease in Koreans]. AB - BACKGROUND/AIMS: Toll-like receptors (TLRs) serve as pattern recognition receptors that recognize specific molecular patterns of pathogens and can mediate the production of proinflammatory cytokines. Recently, TLRs have been identified as susceptibility genes for Crohn's disease (CD) in several studies from Western populations. We investigated the association of genetic variations in TLR4 and TLR9 with CD in Korean population. METHODS: In 380 CD cases and 380 healthy controls, we performed genotyping for TLR4 Asp299Gly (rs4986790) and Thr399Ile (rs4986791). The genetic variations in the TLR9 -1237T/C (rs5743836) were also examined. RESULTS: Among CD patients genotyped for TLR4 Asp299Gly and TLR9 1237T/C, none had variant alleles. Similarly, none of the subjects genotyped for TLR4 Thr399Ile showed genetic variations. CONCLUSIONS: Our results indicate that the major genetic variations in TLR4 and TLR9 are rare and may not be associated with susceptibility to CD in Koreans. PMID- 20026893 TI - [Comparison of effects of preoperative stenting for obstructing colorectal cancers according to the location of the obstructing lesion]. AB - BACKGROUND/AIMS: With the development of self-expanding metallic stents, colonic obstruction can be relieved without the need for surgery. The results of preoperative placement of stents for malignant colorectal obstruction might be different according to the obstructing lesion. The objective of this study was to compare clinical improvement rates and operative results after preoperative placement of stents for malignant colorectal obstruction according to the location of the obstructing lesion. METHODS: This is a retrospective study including 57 patients who underwent self-expanding metallic stent insertion for obstructing resectable colorectal cancers. Patients were classified into three groups according to the location of the lesion as follows: proximal to the sigmoid colon (Group A), sigmoid colon (Group B), and rectum (Group C). RESULTS: The number of patients in A, B, and C groups was 13, 22, and 22, respectively. No significant differences in age, gender, stent type, and accompanying diseases among the three groups were observed. There were no significant differences in stent-related complications, clinical improvement rates, and one-stage resection rates among the three groups. The postoperative complications, the requirement rate of ICU care, the period of ICU stay, postoperative hospital stay, and hospital mortality did not significantly differ among the three groups. CONCLUSIONS: Clinical improvement rates and operative results after successful placement of stents for obstructing resectable colorectal cancers are not different according to the location of the obstructing lesion, suggesting that preoperative stenting for one-stage curative resection is useful, irrespective of the location of lesion. PMID- 20026894 TI - [The safety assessment of percutaneous transhepatic transpapillary stent insertion in malignant obstructive jaundice: regarding the risk of pancreatitis and the effect of preliminary endoscopic sphincterotomy]. AB - BACKGROUND/AIMS: Metal stent insertion through percutaneous transhepatic biliary drainage (PTBD) track is an important palliative treatment modality for malignant biliary obstruction. Acute pancreatitis is one of serious complications of biliary metal stenting. The purpose of this study was to investigate the risk of pancreatitis for patients who underwent metal stent insertion via PTBD track. METHODS: A retrospective analysis of 90 consecutive patients who received metal stent insertion via PTBD track from Jan. 2002 to Dec. 2007 was carried out. Patients were divided into the transpapillary and non-transpapillary group, and the risks of pancreatitis were compared. The effects of preliminary endoscopic sphincterectomy (EST) was also investigated in transpapillary group. RESULTS: The rate of pancreatitis was higher in transpapillary group compared to nontranspapillary group (odd ratio 1.87, 95% CI 0.516-6.761), but it showed no stastically significance (p=0.502). In transpapillary group, patients who received preliminary EST showed lower rate of pancreatitis (odd ratio 0.91, 95% CI 0.656-1.273), but it showed no stastically significance (p=0.614). CONCLUSIONS: Metallic stent insertion through the intact sphincter of Oddi might have a risk of developing pancreatitis. Further study is needed to elucidate the mechanism of pancreatitis and the way of prevention. PMID- 20026895 TI - [A case of spontaneous pneumoperitoneum associated with idiopathic intestinal pseudoobstruction]. AB - Pneumoperitoneum, free intra-abdominal air, usually results from the perforation of a hollow viscous. In approximately 10% of cases, however, pneumoperitoneum is not caused by gastrointestinal perforation. These cases of "spontaneous pneumoperitoneum" generally follow more benign course and may not require surgical intervention. Examples include cardiopulmonary resuscitation (CPR), malrotation, mechanical ventilator support, gynecologic manipulation, blunt abdominal trauma, and chronic intestinal pseudoobstruction in infancy (Sieber syndrome). But, it is extremely rare of spontaneous pneumoperitoneum secondary to idiopathic intestinal pseudoobstruction in adult. We herein report a patient with chronic idiopathic intestinal pseudoobstruction who developed a pneumoperitoneum. PMID- 20026896 TI - [A case of sclerosed hemangioma mimicking intrahepatic cholangiocarcinoma]. AB - Hemangioma is one of the most frequently encountered benign hepatic neoplasm which can develop secondary degeneration. Sclerosed hemangioma is a rare disease histologically characterized by large amount of collagen and elastic fibril between sclerosed small vessels. Its differential diagnosis is very difficult. It should be included in the differential diagnosis of other hepatic lesions such as hepatocellular carcinoma, intrahepatic cholangiocarcinoma, and metastatic hepatic tumor. A 77-year old male was admitted with upper abdominal discomfort. Abdominal ultrasonography revealed GB stone, dilated common bile duct with bile duct stone, and a 4.6 cm sized hyperechoic mass at segment 5 and 6 of the liver. Abdominal dynamic computed tomography demonstrated dilated intrahepatic bile ducts and a 5 x 5 cm sized mass which showed minimally delayed enhancement. Abdominal magnetic resonance imaging revealed the mass with low signal intensity in T1 weighted image, high signal intensity and focal low signal in T2 weighted image which showed minimal enhancement. We removed common bile duct stone with endoscopic retrograde cholangiopancreatography then decided to undergo right lower segmentectomy of liver due to possibility of cholangiocarcinoma. Histopathological examination of hepatic mass showed large amount of fibrous tissue with occasional residual vascular channels. We describe one case of sclerosed hemangioma mimicking cholangiocarcinoma. PMID- 20026897 TI - [A case of congenital hepatic fibrosis presented with recurrent acute cholangitis]. AB - Acute cholangitis usually develops in congenital hepatic fibrosis (CHF), accompanied by cystic dilated bile ducts. However, it can also develop in simple CHF and may lead to critical course. A 30-year old man presented with recurrent acute cholangitis without bile duct dilatation. He visited the hospital for febrile sense and abdominal pain in the right upper quadrant. He had been admitted several times for hepatosplenomegaly and cholangitis since childhood and received a liver biopsy 15 years ago. Abdominal computed tomography (CT) and endoscopic retrograde cholangiopancreatography (ERCP) revealed hepatosplenomegaly and a mildly dilated bile duct without stones or biliary cysts. His condition improved after conservative treatment. However, during a two-month follow up period, the patient experienced three episodes of acute cholangitis. A liver biopsy was performed and showed periportal fibrosis and intrahepatic ductular dysplasia, characteristics of congenital hepatic fibrosis. The periportal fibrosis and the infiltration of inflammatory cells were aggravated compared to 15 years ago. There was no evidence of hepatic cirrhosis. He was diagnosed with congenital hepatic fibrosis with recurrent acute cholangitis without intrahepatic duct dilatation, and conservatively treated with antibiotics. PMID- 20026898 TI - [Paraganglioma of the pancreas metastasized to the adrenal gland: a case report]. AB - Paraganglioma is a rare neuroendocrine tumor arising from the neural crest, which includes tissues such as the adrenal medulla, carotid and aortic body, organs of Zuckerkandl, and other unnamed paraganglia. The head, neck, and retroperitoneum are the most common sites for paraganglioma. However, paraganglioma of the pancreas is extremely rare. We report our experience of this rare disease. A 70 year old female patient admitted for a pancreas tail mass detected by computed tomography (CT) scan checked for vague left flank pain. CT with contrast enhancement showed a 4.2-cm heterogeneously enhanced lesion in the tail of the pancreas. A well defined ovoid shape mass in left adrenal gland was suggested adenoma. Distal pancreatectomy with left adrenalectomy was performed. Any lymph node enlargement was not found. Pancreas mass did not invade adjacent organ. Microscopic examination with pancreas and adrenal gland revealed that the cells were arranged in a characteristic Zellballen pattern. Immunohistochemical staining revealed positivity for neuron-specific enolase, chromogranin A, synaptophysin, and S-100 protein. On the basis of these findings, we diagnosed the tumor as a paraganglioma of the pancreas and adrenal gland. We report the first case of pancreas paraganglioma in Korea. PMID- 20026899 TI - [Effect of stenting for obstructing colorectal cancers, according to the location of obstructing lesion]. PMID- 20026900 TI - Analysis of HIF-1a and its regulator, PHD2, in retroperitoneal sarcomas: clinico pathologic implications. AB - Hypoxia is known to play important role in cancer biology. In sarcomas, hypoxia induced protein biomarkers such as Hypoxia Inducible Factor-1a (HIF-1a), vascular endothelial growth factor (VEGF), and Erythropoietin (Epo) have been previously reported in only a few studies. Moreover, the biologic significance and relationship to tumorigenesis of these hypoxia-induced biomarkers is not well understood in the context of sarcoma. The HIF negative regulator, Prolyl Hydroxylase Domain protein 2 (PHD2) has not been evaluated in sarcomas. We examined the expression of PHD2, HIF-1a, and several other hypoxia induced biomarkers in a series of clinically characterized, retroperitoneal sarcomas with immunohistochemical methods. Expression of these proteins was analyzed and correlated with clinical outcome. Increased HIF-1a expression was associated with shorter overall and disease free survival. PHD2 expression was detected in the majority of sarcoma cases, with increased expression correlating with high tumor grade but not with survival. Though changes in PHD2 expression alone did not correlate with overall and disease free survival, reduced/absent PHD2 expression in the presence of HIF-1a expression was associated with shorter overall and disease-free survival than that of other HIF-1a/PHD2 expression profiles. These observations suggest that regulation and expression of both PHD2 and HIF-1a are important to the biology of sarcomas, and that loss of PHD2 function has an additional adverse effect in the prognosis of sarcomas in tumors expressing HIF-1a. The biologic and therapeutic implications of HIF-1a and PHD2 expression in retroperitoneal sarcomas warrant further investigation. PMID- 20026901 TI - Neuropilin, you gotta let me know: should I stay or should I go? AB - Neuropilins are highly conserved single pass transmembrane proteins specific to vertebrates. They were originally identified as adhesion molecules in the nervous system, but were subsequently rediscovered as the ligand binding subunit of the class 3 semaphorin receptor in neurons and then as blood vessel receptors for the vascular endothelial growth factor VEGF. More recently they have also been implicated as mediators of the T-cell immune response and as key prognostic markers in several types of cancer. Because neuropilins bind multiple ligands and associate with several different types of co-receptors, they variably promote cell adhesion, repulsion or attraction. Which response they ultimately invoke is decided by the cellular and even subcellular context the neuropilins find themselves in. Here, we review how the developmental functions of the neuropilins are influenced by such different contexts. PMID- 20026902 TI - Traditional and new prognosticators in breast cancer: Nottingham index, Mib-1 and estrogen receptor signaling remain the best predictors of relapse and survival in a series of 289 cases. AB - Histopathological and immunohistochemical findings on tissue microarrays, overall survival (OS), disease-free survival (DFS) and incidence of relapses (R) were recorded and statistically analyzed in 289 breast cancers. A higher R and a shorter DFS were significantly related to larger tumors, lymph node invasion, higher tumor grade, absence of estrogen receptors (ER), triple negative tumors, and presence of lymphovascular invasion (LVI). Longer OS was observed to be significantly associated with smaller tumor size (T), lymph node negativity, lower tumor grade, absence of LVI, lower Mib-1 expression and with the presence of ER. At multivariate analysis, only T for DFS and lymph node status and triple negativity either for DFS or OS had independent prognostic value. In the 194 lymph node-negative women DFS and OS were inversely related to tumor grade, absence of ER, Mib-1 expression in more than 15% of neoplastic cells and, only for DFS, presence of LVI. In the 95 lymph node-positive the number of involved nodes was the most discriminating parameter either for DFS or OS; T, Her-2 status and presence of LVI were significantly related to DFS. ER negativity was related to higher grade, progesterone receptors (PR) negativity, Her-2 negativity, hence to triple negativity, to basal-like type, Mib-1expression over 15% of neoplastic cells. Her-2 positivity was related to higher grade, ER positivity and PR positivity. Basal-like type was not an independent prognosticator, while triple negative type has a significant relation to shorter OS. The Nottingham prognostic index accurately identifies prognostic groupings and Mib-1 expression and ER signaling are the key biological predictors even in single cases. PMID- 20026903 TI - Over-expression of Nanog predicts tumor progression and poor prognosis in colorectal cancer. AB - We studied the expression and regulatory effects of ESC self-renewal molecule Nanog in colorectal cancer (CRC). Immunohistochemical analysis of 175 colorectal tumor samples showed that overexpression of Nanog was strongly correlated with poor prognosis, lymph node metastasis and Dukes classification for CRC. Univariate and multivariate survival analyses further indicated that Nanog expression was a potential prognostic factor for CRC. Gain-of-function analysis revealed that lentivirus-mediated Nanog overexpression promoted proliferation, motility and migration of human CRC cells. Interestingly, we found that Nanog played as both an inducer and a receipt of epithelial-mesenchymal transition (EMT) related signals. Nanog induced expression of Slug and Snail, two major regulator of EMT. Meanwhile, Nanog could also be regulated by Snail and initiated by TGF-beta1. Our data demonstrate self-renewal gene Nanog has a prognostic role in CRC, which functions in progression of CRC by promoting proliferation, invasion, and motility of human CRC cells, and participates EMT process during CRC progression. PMID- 20026904 TI - Over-expression of angiotensin II type 2 receptor gene induces cell death in lung adenocarcinoma cells. AB - The endogenous angiotensin II (Ang II) type 2 receptor (AT 2) has been shown to mediate apoptosis in cardiovascular tissues. Thus, the aim of this study was to explore the anti-cancer effect of AT 2 over-expression on lung adenocarcinoma cells in vitro using adenoviral (Ad), FuGENE, and nanoparticle vectors. All three gene transfection methods efficiently transfected AT 2 cDNA into lung cancer cells but caused minimal gene transfection in normal lung epithelial cells. Ad-AT 2 significantly attenuated multiple human lung cancer cell growth (A549 and H358) as compared to the control viral vector, Ad-LacZ, when cell viability was examined by direct cell count. Examination of annexin V by flow cytometry revealed the activation of the apoptotic pathway via AT 2 over-expression. Western Blot analysis confirmed the activation of caspase-3. Similarly, poly (lactide-co-glycolic acid) (PLGA) biodegradable nanoparticles encapsulated AT 2 plasmid DNA were shown to be effectively taken up into the lung cancer cell. Nanoparticle-based AT 2 gene transfection markedly increased AT 2 expression and resultant cell death in A549 cells. These results indicate that AT 2 over expression effectively attenuates growth of lung adenocarcinoma cells through intrinsic apoptosis. Our results also suggest that PLGA nanoparticles can be used as an efficient gene delivery vector for lung adenocarcinoma targeted therapy. PMID- 20026905 TI - Targeting plasma membrane GRP78 for cancer growth inhibition. PMID- 20026906 TI - Cell culture-induced aberrant methylation of the imprinted IG DMR in human lymphoblastoid cell lines. AB - DNA methylation patterns are often poorly conserved through cell culturing. To determine the effect of cell immortalization and culture on DNA methylation profiles, we analyzed methylation in the differentially methylated regions (DMR) of five imprinted domains: the intergenic (IG) DMR on chromosome 14q32; potassium voltage-gated channel, KQT-like subfamily, member 1, (KCNQ1); small nuclear ribonucleoprotein polypeptide N (SNRPN), mesoderm specific transcript homolog (MEST); and H19 in lymphoblastoid cell lines (LCLs). In the IG DMR we found an aberrant methylation pattern that was consistent through all the cell lines tested and significantly different from that of noncultured peripheral blood cells. Using a generalized linear mixed model to compare methylation profiles, we show that recently derived LCLs significantly differ from the CEPH LCLs. This implies a gradual cell-culture related deterioration of DNA methylation in the IG DMR with at least two steps that may be identified: loss of methylation at CG sites 1 and 8; and loss of allelic differences in DNA methylation. The IG DMR methylation profile also confirms the high level of clonality of the CEPH LCLs. We conclude that non-transformed primary cells may be less susceptible to epigenetic anomalies and therefore may provide a more accurate reflection of gene expression in vivo. PMID- 20026907 TI - Definitive endoderm derived from human embryonic stem cells highly express the integrin receptors alphaV and beta5. AB - Human embryonic stem cells (hESCs) can be directed to differentiate into a number of endoderm cell types, however mature functional cells have yet to be produced in vitro. This suggests that there may be important factors that have yet to be described, which may be essential for the proper derivation of these cells. One such factor is the integrin mediated interactions between a cell and the extracellular matrix (ECM). On this basis, the present study investigated the role of the ECM in the directed differentiation of hESCs to definitive endoderm via analysis of integrin gene expression. The results showed that definitive endoderm can be efficiently and effectively derived from hESCs in a feeder free, single defined ECM of laminin. Analysis of integrin expression also showed that definitive endoderm highly express the integrins alphaV and beta5, which have the ability to bind to vitronectin, whilst expression of the pluripotency related laminin binding integrins alpha3, alpha6 and beta4 were downregulated. This suggested a potential role of vitronectin binding integrins in the development of definitive endoderm. PMID- 20026908 TI - The p300/CBP associated factor is frequently downregulated in intestinal-type gastric carcinoma and constitutes a biomarker for clinical outcome. AB - OBJECTIVES: To investigate the expression of P300/CBP-associated factor (PCAF) protein in intestinal type gastric cancer (ITGC); analyze the relationship between the expression of PCAF protein and the clinical pathological characteristics of patients; explore the effects of PCAF protein on biological behaviors of ITGC. RESULTS: The expression of PCAF was markedly down-regulated in GC cell lines and ITGC tissues. PCAF was able to suppress tumorigenicity of GC cells both in vitro and in vivo, including colony formation in soft agar and tumor formation in nude mice. PCAF could also inhibit GC cells entering S phase from G1 phase. Statistical analysis displayed a significant correlation in PCAF expression with the gastric wall invasion, tumor size, TNM stage, p21, pRb (P<0.001) and PCNA (P<0.01) in ITGC specimens. A reduced PCAF protein expression correlated significantly with a mutant type p53 protein expression (P<0.01). Univariate analysis indicated that the patients demonstrating the high-PCAF/wild type p53 expression have a significantly (P<0.0001) better overall survival (OS), while multivariate analysis indicated that the location, lymph node metastasis, PCAF/p53 (P<0.0001), gastric wall invasion (P=0.001) and PCNA (P=0.018) are independently significant prognostic factors for OS. METHODS: Immunohistochemistry was performed to evaluate the expression of PCAF in a large subset containing 406 ITGC samples. Eukaryotic expression plasmid pcDNA3.1/PCAF was constructed and transfected into the human gastric cancer cell line SGC-7901 and protein expression was detected by Western blot. The proliferation and cell cycle of gastric cancer cells were evaluated by MTT assay and flow cytometry. Tumor growth in nude mice was used to access the tumorigenicity of gastric cancer cells. Apoptosis cells were detected by TUNEL staining. CONCLUSION: Reduced expression of PCAF plays an important role in the development of ITGC and correlates with a poor clinical outcome. PMID- 20026909 TI - MYC activity mitigates response to rapamycin in prostate cancer through 4EBP1 mediated inhibition of autophagy. AB - Cancer cells have evolved exquisitely to ignore both intrinsic and extrinsic cell death signals, and resistance to cell death is a critical challenge facing clinical oncology. Autophagy, the catabolic recycling process that involves the fusion of autophagosomes containing sequestered cargo with lysosomes, has an enigmatic role in tumorigenesis. In times of metabolic stress due to deprived nutrition or hypoxia, tumor cells use autophagy as a scavenging mechanism for maintenance of critical processes and survival. However, modulation of the extent of autophagy plays a critical role, as excessive autophagy can result in a nonapoptotic and non-necrotic cell death (sometimes referred to as Type II programmed cell death). It is likely that the genetic context of specific cancers will have an impact upon whether autophagy is primarily a mechanism for survival or cell death. PMID- 20026910 TI - A second report from the EMBO conference on autophagy: mechanism, regulation and selectivity of autophagy. AB - Some key questions being examined in the field of autophagy concern the origin of the membrane that forms the sequestering vesicle, the function of the related machinery, including the identification of new components and binding partners of previously identified autophagy-related proteins and the mechanism of autophagic selectivity. Another general area of intense focus pertains to the physiological role of autophagy and its connection with various pathologies including neurodegeneration, muscle wasting and mitochondrially related diseases. PMID- 20026911 TI - Modulation of the SecY channel permeability by pore mutations and trivalent cations. AB - The SecY channel serves to transport proteins across the bacterial inner membrane. The closed channel is impermeable to small molecules by means of a plug domain and a hydrophobic pore, consisting of six conserved isoleucine residues. The substitution of these isoleucines by asparagine leads to the selective conductance of small monovalent anions, especially chloride. In this addendum, we show that replacement of the isoleucine residues by bulky phenylalanine also leads to an increased chloride conductance, suggesting that hydrophobicity of the pore is not the sole determinant for maintaining channel impermeability. Instead, incubation of the membrane with the trivalent cation Al(3+) dramatically increases Cl(-) transport across the wild type SecY channel, suggesting that surface charge density around the SecY pore plays a significant role during the process of chloride conductance. PMID- 20026912 TI - Convergent signaling events underlie dopamine-mediated inhibition of CaV3.2 T type calcium channels. PMID- 20026913 TI - Swapping the I-II intracellular linker between L-type CaV1.2 and R-type CaV2.3 high-voltage gated calcium channels exchanges activation attributes. AB - Calcium entry through voltage-gated calcium channels (VGCC) initiates diverse cellular functions. VGCC pore-forming subunit (Ca(V)alpha(1)) contains four homology repeats, each encompassing a voltage sensor and a pore domain. Three main classes of Ca(V)alpha(1) subunits have been described, Ca(V)1, Ca(V)2 and Ca(V)3 that differ in their voltage-dependence of activation and in the extent in which this process is modulated by the auxiliary beta-subunit (Ca(V)beta). Association of Ca(V)beta induces a coil-to-helix conformation of the I-II intracellular linker joining the first and second repeat of Ca(V)alpha(1) that is thought to be crucial for modulation of channel function. When expressed in Xenopus laevis oocytes in the absence of Ca(V)beta, the voltage to reach 50% activation (V(0.5)) for Ca(V)1.2 and Ca(V)2.3 differs by more than 60 mV and the channel current-carrying capacity by more than thirty-fold. Here we report that the difference in V(0.5) is reduced to about 30 mV and the current-carrying capacity becomes virtually identical when the I-II linkers of Ca(V)1.2 and Ca(V)2.3 are swapped. Co-expression with Ca(V)beta increases the current-carrying capacity of chimeric channels by the same extent, while the difference in V(0.5) with respect to their corresponding parental channels vanishes. Our findings indicate that Ca(V)beta modulatory potency is determined by both, the nature of the I-II linker and the pore-forming subunit background. Moreover, they demonstrate that the I-II linker encodes self-reliant molecular determinants for channel activation and suggest that besides the secondary structure adopted by this segment upon Ca(V)beta association, its chemical nature is as well relevant. PMID- 20026914 TI - Clinical symposium's founder defines leadership. PMID- 20026919 TI - Beware of inappropriately billing clinic visits/evaluation and managements with procedures. PMID- 20026920 TI - The use of acticoat moisture control for the successful treatment of diabetic ulcers. PMID- 20026921 TI - Reducing hospital-acquired pressure ulcers with a silicone-based dermal nourishing emollient-associated skincare regimen. AB - OBJECTIVE: To determine the effect of a silicone-based dermal nourishing emollient (SBDNE) regimen on the reduction of pressure ulcers (PrUs) and costs in a hospital medical unit. RATIONALE: PrUs represent a serious problem for patients within the acute care setting and are a significant care management challenge for clinicians. Effective October 1, 2008, the Centers for Medicare and Medicaid Services will no longer reimburse hospitals at the higher diagnosis-related group rate for Stages III and IV PrUs that are not documented on admission. In addition, formation of PrUs in the hospital also puts the institution at financial risk of lawsuits. RESEARCH DESIGN AND METHODS: The wound healing center at Porter Adventist Hospital, Denver, Colorado, documented the hospital-acquired incidence rate of PrU patients in the hospital from May 2006 to December 2007. A retrospective, quasi-experimental design was used to examine the changes in PrU incidence rates and the economic effect of introducing a SBDNE regimen into an existing PrU prevention protocol. RESULTS: The replacement of a mixture of ad hoc skin care products, none of which contained silicone-based emollients, with an SBDNE skin care regimen into an existing prevention program significantly reduced the proportion of hospital-acquired PrUs to 0% after 8 months. Estimated cost savings per patient admitted to the medical unit attributed to SBDNE averaged $6677.11 per patient. MAIN CONCLUSIONS: The use of an SBDNE skin care regimen was important in bringing about a significant reduction in the number of patients with PrUs and respective treatment costs in a medical unit experiencing high incidence rates of PrUs. PMID- 20026922 TI - The Toronto Symptom Assessment System for Wounds: a new clinical and research tool. AB - OBJECTIVE: To formulate a patient-rated assessment tool that facilitates the measurement of pain and polysymptom distress directly related to all classes of wounds. DESIGN: A prospective observational study derived from a sequential case series of patients with advanced illness was carried out to determine the most common symptoms associated with wounds from 9 distinct classes (malignant, pressure ulcers, iatrogenic, traumatic, diabetic foot ulcers, venous ulcers, arterial ulcers, infections/inflammatory lesions, and ostomies). Ten wound related symptoms were identified and used to create a patient-scored assessment tool. The Toronto Symptom Assessment System for Wounds (TSAS-W) was then developed and used in a pilot trial during which patients completed TSAS-W at baseline and 7 days later. PARTICIPANTS: Five hundred thirty-one patients either presented with wounds at baseline or developed them during the 24-month follow-up period. Patients affected by any type of wound were asked to report on the top 3 symptoms directly attributable to their wounds. The pilot trial of TSAS-W involved 103 wounds afflicting 83 sequential patients. MAIN RESULTS: The most prevalent wound-related symptoms included pain, exudation, odor, itching, bleeding, aesthetic concern, swelling, and mass and bulk effects from the wound and associated dressings; 78.6% of the TSAS-W assessments were carried out by the patient alone, 14.6% were carried out by the patient assisted by a caregiver, and 6.8% were carried out entirely by a caregiver. The summation of all 10 TSAS-W parameters, the global wound symptom distress score (GWSDS), resulted in a mean for all wounds of 34.47 at baseline and decreased to a mean of 28.40 at 7 days later. Cosmetic or aesthetic concern and/or distress was associated with the highest mean scores of all symptoms. Malignant wounds and wounds involving the perineum and genitalia were associated with the highest GWSDSs. CONCLUSION: The TSAS-W is a new tool for systematically assessing the degree of pain and polysymptom distress associated with all classes of wounds. It is modeled after the Edmonton Symptom Assessment System that is widely used and validated in the palliative care arena. TSAS-W is composed of 10 symptom parameters that are individually assessed on 11-point numeric rating scales (0-10). The summation of all of the element symptom scores equates to a GWSDS. It may be used in the clinical setting to guide wound-related pain and polysymptom management. In addition, TSAS-W may be useful as a tool in facilitating clinical audit and future wound care research. PMID- 20026923 TI - Annual checkup: the CMS pressure ulcer present-on-admission indicator. AB - PURPOSE: To provide the wound care practitioner with information about the Centers for Medicare and Medicaid Services (CMS) pressure ulcer present-on admission (POA) indicators. TARGET AUDIENCE: This continuing education activity is intended for physicians and nurses with an interest in skin and wound care. OBJECTIVES: After reading this article and taking this test, the reader should be able to: 1. Describe the history and rationale for the CMS financial incentives for pressure ulcer (PrU) prevention. 2. Identify the CMS Pressure Ulcer POA indicators and 2009 ICD-9 coding for PrU staging. 3. Discuss the implications of the new CMS reimbursement for PrUs on healthcare organization practices with examples of successful interventions for PrU reduction. PMID- 20026925 TI - Driving documentation into the future. PMID- 20026926 TI - Ghostwriters in the fray: the implications for science, industry, medical journals, and our patients. PMID- 20026930 TI - CMS limits consignment closets and stock and bill arrangements. PMID- 20026931 TI - Herpes zoster affecting the lower extremity. PMID- 20026932 TI - Contribution of skin temperature regularity to the risk of developing pressure ulcers in nursing facility residents. AB - OBJECTIVES: The purpose of the study was to determine whether characterizing skin temperature regulation as a functional property of the skin as it relates to tissue tolerance improves the clinician's understanding of pressure ulcer risk prediction. DESIGN: A 2-group time-series design was used to observe skin temperature regularity (entropy) and self-similarity (spectral exponent). METHODS: Twenty nursing facility residents wore skin temperature monitors continuously for 5 days. One bathing episode was observed because bathing is a commonly occurring care procedure. PRIMARY OUTCOME MEASURE: Difference in skin temperature multiscale entropy and spectral exponent by risk category and pressure ulcer outcome. RESULTS: Multiscale entropy (MSE) for skin temperature was lowest in those who developed pressure ulcers, F1,18 = 35.14, P < .001. Skin temperature mean MSE, F1,17 = 5.55, P = .031 and the skin temperature spectral exponent, F1,17 = 6.19, P = .023 differentiated the risk groups. The change in skin temperature entropy during bathing was significant, t(16) = 2.55, P = .021. CONCLUSIONS: Skin temperature MSE and the spectral exponent were significantly different between low-risk and higher risk residents and residents who did and did not develop pressure ulcers. The study supports measurement of skin temperature regulation as a component of tissue tolerance to pressure. PMID- 20026933 TI - Promote pressure ulcer healing in individuals with spinal cord injury using an individualized cyclic pressure-relief protocol. AB - OBJECTIVE: To evaluate whether an individualized cyclic pressure-relief protocol accelerates wound healing in wheelchair users with established pressure ulcers (PrUs). DESIGN: Randomized controlled study. SETTING: Spinal cord injury clinics. PARTICIPANTS: Forty-four subjects, aged 18-79 years, with a Stage II or Stage III PrU, were randomly assigned to the control (n = 22) or treatment (n = 22) groups. INTERVENTIONS: Subjects in the treatment group used wheelchairs equipped with an individually adjusted automated seat that provided cyclic pressure relief, and those in the control group used a standard wheelchair. All subjects sat in wheelchairs for a minimum of 4 hours per day for 30 days during their PrU treatment. MAIN OUTCOME MEASURES: Wound characteristics were assessed using the Pressure Ulcer Scale for Healing (PUSH) tool and wound dimensions recorded with digital photographs twice a week. Median healing time for a 30% healing relative to initial measurements, the percentage reduction in wound area, and the percentage improvement in PUSH score achieved at the end of the trial were compared between groups. RESULTS: At the end of 30 days, both groups demonstrated a general trend of healing. However, the treatment group was found to take significantly less time to achieve 30% healing for the wound measurement compared with the control group. The percentage improvement of the wound area and PUSH scores were greater in using cyclic seating (45.0 +/- 21.0, P < .003; 29.9 +/- 24. 6, P < .003) compared with standard seating (10.2 +/- 34.9, 5.8 +/- 9.2). CONCLUSIONS: The authors' findings show that cyclically relieving pressure in the area of a wound for seated individuals can greatly aid wound healing. The current study provides evidence that the individualized cyclic pressure-relief protocol helps promote pressure wound healing in a clinical setting. The authors concluded that the individualized cyclic pressure relief may have substantial benefits in accelerating the healing process in wheelchair users with existing PrUs, while maintaining the mobility of individuals with SCI during the PrU treatment. PMID- 20026934 TI - Peristomal skin complications and management. AB - PURPOSE: : To provide the wound care practitioner with an overview of practical approaches to prevent and treat common peristomal skin conditions. TARGET AUDIENCE: : This continuing education activity is intended for physicians and nurses with an interest in skin and wound care. OBJECTIVES: : After participating in this educational activity, the participant should be better able to: PMID- 20026936 TI - Part 1: documentation steps for success. PMID- 20026938 TI - Positive mental aging. PMID- 20026940 TI - Dementia diagnosis. PMID- 20026942 TI - Open gastrostomy for noninvasive ventilation users with neuromuscular disease. AB - OBJECTIVE: To report open gastrostomy for ventilator-assisted or -supported patients with altered nutritional status as a result of severe dysphagia and without tracheostomy, translaryngeal intubation, or general anesthesia. Avoiding intubation and general anesthesia decreases the risk of respiratory complications and can prolong noninvasive respiratory management. DESIGN: The procedure was performed for 62 noninvasive intermittent positive-pressure ventilation users: 44 with amyotrophic lateral sclerosis, 10 with muscular dystrophy including 6 with Duchenne muscular dystrophy, and 8 with other conditions. All had vital capacities <40% of predicted normal. Noninvasive intermittent positive-pressure ventilation was provided in ambient air before, during, and after the procedure. Oxyhemoglobin saturation was maintained at 95% or greater and end-tidal CO2 <40 cm H2O by noninvasive intermittent positive-pressure ventilation and mechanically assisted coughing. RESULTS: There were no complications of the procedure. All patients gained weight subsequently. Mean postgastrostomy survival was 38.8 +/- 6.2 mos with 26 of the patients still alive. Eighteen of the 62 patients had no ventilator-free breathing ability before, during, or after the gastrostomy. Fifty one patients eventually lost all ventilator-free breathing abilities without tracheostomy. CONCLUSIONS: Open gastrostomy can be performed safely without airway intubation or general anesthesia for patients with little or no autonomous breathing ability. It permitted continued survival without tracheostomy despite loss of all ventilator-free breathing abilities for 51 patients. PMID- 20026943 TI - Walking index for spinal cord injury version 2 (WISCI-II) with repeatability of the 10-m walk time: Inter- and intrarater reliabilities. AB - OBJECTIVE: To demonstrate the inter-/intrarater reliability of the Walking Index for Spinal Cord Injury version 2 scale and the repeatability of the time to walk 10 m in chronic subjects. DESIGN: In this reliability study, 26 subjects from the United States and Italy with spinal cord injury/disorder were tested by two blinded raters on two separate days to determine self-selected and maximum Walking Index for Spinal Cord Injury levels and the time to complete a 10-m walk. Subjects were progressed from self-selected to maximum Walking Index for Spinal Cord Injury incrementally until they failed the higher level. Intraclass correlations were calculated for Walking Index for Spinal Cord Injury levels and repeatability coefficients for the 10-m time. RESULTS: Twenty-two of 26 subjects showed increases of one to eight levels from self-selected to maximum Walking Index for Spinal Cord Injury, whereas 10-m walking time remained relatively unchanged (n = 15) or increased markedly (n = 7). Inter- and intrarater reliabilities were 1.00 for the self-selected Walking Index for Spinal Cord Injury level. Intrarater reliability for the maximum level was 1.0; interrater reliability was 0.98. Repeatability coefficients for time to walk 10 m were smaller (better) at self-selected than at maximum Walking Index for Spinal Cord Injury and on the same day than on different days. On same-day assessments, repeatability coefficients were 18%-20% of 10-m walk time, excluding subjects with discrepant Walking Index for Spinal Cord Injury levels (n = 2). For different-day assessments, repeatability coefficients were 27%-35% of 10-m walk time. CONCLUSIONS: The determination of both self-selected and maximum Walking Index for Spinal Cord Injury levels is highly reliable, whereas 10-m walking time is more variable. Walking "profiles" of speed at self-selected and maximum Walking Index for Spinal Cord Injury may better characterize walking ability than a single Walking Index for Spinal Cord Injury level. PMID- 20026944 TI - Age-related differences in physiologic and psychosocial outcomes after cardiac rehabilitation. AB - OBJECTIVE: To examine differences in physiologic and psychosocial outcomes between age groups after an exercise-based supervised-recovery phase II cardiac rehabilitation outpatient program. DESIGN: This is a longitudinal observational study. The study assessed 442 consecutive cardiac patients. Patients were divided into the middle-aged group (<65 yrs, n = 242) and older-age group (> or =65 yrs, n = 200). Peak oxygen uptake, handgrip and knee extensor muscle strength, upper- and lower-body self-efficacy for physical activity, and physical component summary and mental component summary scores as assessed by SF-36 were measured at 1 and 3 mos after the onset of acute myocardial infarction or cardiac surgery and were compared. RESULTS: All physiologic and psychosocial outcomes increased significantly between months 1 and 3 in both groups. However, increases were greater in the middle-aged vs. older-aged group in peak oxygen uptake (+13.1% vs. +8.7%, P < 0.01), knee extensor muscle strength (+17.6% vs. +13.3%, P = 0.01), lower-body self-efficacy for physical activity (+17.3% vs. +12.7%, P = 0.02), and physical component summary score (+5.4% vs. +2.7%, P = 0.02). CONCLUSIONS: Age related differences in various physiologic and psychosocial measures indicated greater improvement from an exercise-based supervised recovery-phase II cardiac rehabilitation outpatient program in middle-aged vs. older-aged patients. Older adults may derive equal mental or emotional benefit from such a cardiac rehabilitation program but do not experience as much improvement in physiologic outcomes as middle-aged adults. PMID- 20026945 TI - The effects of increased prosthetic ankle motions on the gait of persons with bilateral transtibial amputations. AB - OBJECTIVE: To determine whether the provision of prosthetic ankle motion improves walking performance in persons with bilateral transtibial amputations. DESIGN: Crossover experimental design in which 19 persons with bilateral transtibial amputations were fitted with Endolite Multiflex Ankles (flexion unit) and Otto Bock Torsion Adapters (torsion unit) to increase relative motion between the prosthetic foot and socket in the sagittal and transverse planes, respectively. Quantitative gait analyses were performed on subjects as they walked with four prosthetic configurations: baseline without flexion or torsion units, with only the flexion unit, with only the torsion unit, and with both the flexion and torsion units. Data were compared with a control group of 14 able-bodied subjects. RESULTS: The flexion unit increased ankle sagittal plane motion (6-7 degrees) and increased positive ankle power (about 0.17 W/kg). The torsion unit increased transverse plane ankle range of motion by 1-2 degrees. Responses from questionnaires indicated that 14 of the 19 subjects preferred the prosthetic configuration that included both the flexion and torsion units. Further, the subjects perceived that the increased prosthetic ankle motion was particularly beneficial for improving stability while they walked on uneven terrain. CONCLUSIONS: Both the subjective and objective results suggest that prosthetic foot and ankle components that allow for greater sagittal and transverse plane rotations provide substantial benefit during walking and should be considered for persons with bilateral transtibial amputations. Nonetheless, clinicians should perform individual and appropriate assessments of patients to ensure that they are capable of using components that may improve mobility while possibly sacrificing some degree of stability. PMID- 20026947 TI - MMPI disability profile is associated with degree of opioid use in chronic work related musculoskeletal disorders. AB - OBJECTIVE: To examine the relationship between level of opioid use and Minnesota Multiphasic Personality Inventory (MMPI) findings among chronic pain patients who were about to begin a functional restoration program. METHODS: A prospective cohort study of patients with chronic disabling occupational musculoskeletal disorders. A total of 768 consecutive patients with valid MMPI were divided into 2 groups: 398 patients who reported no opioid use upon admission (No); and 370 patients who reported opioid use upon admission (Yes). Average daily opioid doses (in morphine equivalents) could be determined for 287 of 370 patients, who were further divided into 4 opioid subgroups: Low (>0 to 30 mg, n=148); Medium (>30 to 60 mg, n=57); High (>60 to 120 mg, n=47); and Very High (>120 mg, n=35). RESULTS: Seventy-five percent of the patients who produced valid MMPI profiles could be classified into 1 of the 4 MMPI profiles. Of those patients who could be classified, approximately 7% showed a Normal profile, 15% showed a Conversion V, 9% showed a Neurotic Triad, and 69% showed the Disability Profile. Although the Disability Profile accounted for the majority of patients in all opioid subgroups, the proportions did increase with pretreatment opioid dose, as expected, indicating a relationship between degree of psychopathology and level of pretreatment opioid use. Patients who did not take pretreatment opioids showed the highest proportions of Conversion V and Normal profiles, which indicate a lesser degree or absence of psychopathology, respectively. Patients who took pretreatment opioids were more than one-and-a-half times as likely as patients who did not take pretreatment opioids to produce the Disability Profile, whereas patients taking very high doses of pretreatment opioids were nearly 3 times as likely to produce this profile as patients who took no pretreatment opioids. CONCLUSIONS: The results of this study support the hypothesis that increasing levels of pretreatment opioid use is associated with less desirable MMPI profiles, specifically the Disability Profile and, thus, greater levels of pretreatment psychopathology. PMID- 20026948 TI - The relation of social functioning to school impairment among adolescents with chronic pain. AB - OBJECTIVES: To further our understanding of social functioning in children with chronic pain, and particularly how social functioning relates to school impairment in this population. METHODS: This study involved 126 adolescents (12 to 17 y) evaluated at a multidisciplinary pain clinic. Adolescents completed measures assessing social functioning, pain, physical limitations, somatic symptoms, and school impairment. RESULTS: Lower social functioning scores were significantly associated with pain, physical limitations, somatic symptoms, and school impairment. Social functioning mediated the relations between adolescents' pain experience (ie, pain, physical symptoms, physical limitations) and school impairment. DISCUSSIONS: These findings highlight the importance of assessing and addressing social functioning in youth with chronic pain. Future research targeting school impairment should include evaluating the potential role that peer difficulties may play. PMID- 20026946 TI - Trends in use of opioids for chronic noncancer pain among individuals with mental health and substance use disorders: the TROUP study. AB - OBJECTIVES: Use of prescription opioids for chronic pain is increasing, as is abuse of these medications, though the nature of the link between these trends is unclear. These increases may be most marked in patients with mental health (MH) and substance use disorders (SUDs). We analyzed trends between 2000 and 2005 in opioid prescribing among individuals with noncancer pain conditions (NCPC), with and without MH and SUDs. METHODS: Secondary data analysis of longitudinal administrative data from 2 dissimilar populations: a national, commercially insured population and Arkansas Medicaid enrollees. We examined these opioid outcomes: (1) rates of any prescription opioid use in the past year, (2) rates of chronic use of prescription opioids (greater than 90 d in the past year), (3) mean days supply of opioids, (4) mean daily opioid dose in morphine equivalents, and (5) percentage of total opioid dose that was Schedule II opioids. RESULTS: In 2000, among individuals with NCPC, chronic opioid use was more common among those with a MH or SUD than among those without in commercially insured (8% vs. 3%, P<0.001) and Arkansas Medicaid (20% vs. 13%, P<0.001) populations. Between 2000 and 2005, in commercially insured, rates of chronic opioid use increased by 34.9% among individuals with an MH or SUD and 27.8% among individuals without these disorders. In Arkansas Medicaid chronic, opioid use increased by 55.4% among individuals with an MH or SUD and 39.8% among those without. DISCUSSION: Chronic use of prescription opioids for NCPC is much higher and growing faster in patients with MH and SUDs than in those without these diagnoses. Clinicians should monitor the use of prescription opioids in these vulnerable groups to determine whether opioids are substituting for or interfering with appropriate MH and substance abuse treatment. PMID- 20026949 TI - Predictors of disability and pain six months after the end of treatment for fibromyalgia. AB - OBJECTIVES: The goal of this study was to identify factors associated with decreased disability and lower pain scores 6 months after a multimodal treatment program for fibromyalgia (FM). METHODS: Forty-six patients with FM were assessed after having participated in a 3-month outpatient program integrating physiotherapy, occupational therapy, nursing, and cognitive-behavior therapy. A physician examined the patients before treatment and patients who completed a battery of psychosocial questionnaires at baseline, during treatment, at the end of treatment, and 3 and 6 months after the end of treatment. Two separate multivariable linear regression models were built to identify predictors of improvements in disability and pain. RESULTS: Two predictors for improvement in disability were found: an increase in self-efficacy for pain during treatment and better general adherence during treatment. Similarly, one predictor for improvement in pain intensity was found: an increase in self-efficacy for pain during treatment. DISCUSSION: Self-efficacy and adherence are 2 modifiable factors that influence disability and pain intensity in FM. These psychosocial factors need to be addressed in FM treatment programs to assist patients in maintaining posttreatment improvements. PMID- 20026950 TI - Back muscle activation patterns in chronic low back pain during walking: a "guarding" hypothesis. AB - OBJECTIVES: To investigate whether patients with chronic low back pain (CLBP) show "guarded" movements during walking. It is hypothesized that guarding will be reflected by increased lumbar muscle activity during all periods of stride and secondary, relatively lesser relaxation during periods of swing compared with double support. Furthermore, it is hypothesized that higher levels of perceived fear and disability are related to increased muscle activity and less relative relaxation. MATERIALS AND METHODS: In a cross-sectional study 63 patients with CLBP and 33 healthy controls walked on a treadmill at 3.8 km/h. Surface electromyography (sEMG) data of the erector spinae were obtained and smoothed rectified sEMG (SRE) values were calculated per period of swing and double support. The ratio of SRE values in swing to double support was used as a measure of relative relaxation (SRE ratio). In addition, the relationship between SRE values, the Roland Morris Disability Questionnaire, and the Tampa Scale for Kinesiophobia was analyzed in patients with CLBP. RESULTS: Mean SRE values were significantly higher in patients with CLBP than in controls both during periods of double support and swing. SRE ratios were not significantly different between groups. Results showed no influence of disability or fear of movement on either SRE values or ratios. DISCUSSION: In patients with CLBP, increased lumbar muscle activity during all periods of stride, with comparable alteration between swing and double support, suggests difficulties with total muscle relaxation. On the basis of this evaluation, it is concluded that patients with CLBP show a guarding mechanism during walking. No relationship is found between perceived fear, disability, and muscle activity. PMID- 20026951 TI - Time course of pain relief in patients treated with radiotherapy for cancer pain: a prospective study. AB - OBJECTIVES: The purpose of this prospective study was to analyze time course of pain relief by radiotherapy for cancer pain. METHODS: A total of 91 patients with painful bone metastases were treated by radiotherapy with a median total dose of 46 Gy. Pain of the irradiated site was assessed using a numerical rating scale (pain score: 0 to 10) once a week from the beginning of radiotherapy (day 1) for about 5 weeks. RESULTS: In time course analysis of the 91 cases, the mean (+/-SD) pain scores at day 5, day 12, day 19, day 26, day 33, and day 40 were 7.8 (+/ 1.6), 5.3 (+/-2.5), 3.5 (+/-2.5), 2.4 (+/-2.5), 1.6 (+/-2.1), and 1.1 (+/-1.9), respectively, and mean pain score was significantly reduced with time from the start of radiotherapy (P<0.001: repeated measure ANOVA). Mean pain score was significantly reduced every week by d33 (P<0.05: d5 vs. d12, d12 vs. d19, d19 vs. d26, and d26 vs. d33). Complete pain relief was obtained in 45/91 (49%) cases, and partial (> or =50%) pain relief was obtained in 83/91 (91%) cases. The mean time to obtain 50% pain relief was 13 days. The mean time to obtain complete pain relief (n=45) was 24 days. Doses of analgesics were reduced in 28/64 (44%) cases at the end of radiotherapy. DISCUSSION: Telling approximate time course of pain relief seems to reduce patients' anxiety, and knowing time course of pain relief seems to be useful to determine optimal dose of analgesics that changes according to the course. PMID- 20026952 TI - Adverse event reporting in acupuncture clinical trials focusing on pain. AB - OBJECTIVES: To review the quality of adverse event reporting for published randomized controlled trials (RCTs) focusing on acupuncture for pain reduction. With the release of the Consolidated Standards of Reporting Trials (CONSORT) in 2001, the quality of published RCTs has improved. To improve reporting on adverse events, CONSORT expanded the section on harms (adverse events) in 2004. This paper evaluates whether the updated harms guidelines have been implemented in RCTs evaluating acupuncture for pain relief. METHODS: Systematic searches were conducted using the following databases: MEDLINE, Allied & Complementary Medicine, Cumulative Index to Nursing & Allied Health Literature, and All EBM Reviews. Each database was searched from 2005 through 2008, corresponding to the availability of the updated harms guideline. RESULTS: Ten studies met the inclusion criteria of this review. Six of the 10 studies mentioned or discussed adverse events. Four of the 6 studies did not detail how adverse events were collected. Only 2 studies discussed how adverse events were assessed. DISCUSSION: On the basis of our findings, acupuncture clinical trials for pain reduction have yet to comprehensively meet CONSORT's guidelines for adverse event reporting. Acupuncture is commonly used by patients experiencing pain and although typically viewed as a benign and minimally invasive therapy, serious adverse events have been reported in the literature. To effectively and comprehensively document and understand these events, routine reporting according to CONSORT's harms guidelines should become the norm. Both science and patients are served by accurately evaluating the safety of acupuncture for patient populations experiencing pain. PMID- 20026953 TI - Effect of inhomogeneous static magnetic field on dental pain in humans. AB - OBJECTIVES: The aim of this study was to evaluate the pain-inhibitory effect of inhomogeneous static magnetic field (SMF, 0 to 192 mT peak-to-peak magnetic flux density and 19 T/m lateral gradient) exposure on dental pain associated with dentine sensitivity by the quantification of sensory and affective aspects. METHODS: (1) 0 to 10 numerical rating visual analogue scale (NRS) in the first minute of dental treatment following 30 minutes SMF exposure (uncontrolled), and (2) tolerance threshold measurement (TTM) with the help of a pulp meter showing values from 0 to 80 in arbitrary units (step width 10 unit) before and after 30 minutes SMF exposure in 2 sessions: SMF and sham exposure (randomized, double blind, placebo-controlled). Altogether 59 adult patients (26 males+33 females) representing 62 cases with a mean age of 43.6 years (73% between 20 and 50 y) participated. RESULTS: SMF failed to significantly reduce pain perception in the NRS group and to enhance tolerance threshold in the TTM group. DISCUSSION: Common dental disorders often involve an inflammatory state in the oral environment. Although the relatively low participant number and the uncontrolled manner in case of the NRS examination did not allow drawing unambiguous consequences, it seemed that SMF did not have an effect on healthy patients. The only potential candidate for an effect was, when an inflammatory situation occurred at (or under) the place, where the SMF exposure was targeted. In our case the male participants with caries responded most positively on the SMF treatment. PMID- 20026954 TI - When the progression of disease lowers opioid requirement in cancer patients. AB - OBJECTIVE: To describe 2 cases in which an abrupt progression of disease compromising pain pathways and inducing some relief of existing pain or limiting drug delivery to central nervous system. METHODS: Case reports which a significant decrease of opioid requirement was reported, either systematically and spinally. RESULTS: The progression of disease produced changes in pain input or limited the effects of opioids given spinally. DISCUSSION: The data presented suggest that physicians should be aware of the possibility that opioid doses have to be reduced, where presumably specific events related to the progression of disease can change the pain syndrome or reduce the delivery of opioids when using particular routes of administration. This problem needs to be recognized and treated appropriately when it occurs. PMID- 20026955 TI - Acute auditory and vestibular symptoms associated with heat and transdermal lidocaine. AB - OBJECTIVES: To describe a case of systemic lidocaine toxicity from the simultaneous use of transdermal patches and a heating pad. METHODS: Case report and discussion. RESULTS: The focus of this case report are the interesting neuro otologic signs of central lidocaine toxicity that occurred after a patient fell asleep on a heating pad with his Lidoderm patches in place. Hearing improvement, tinnitus reduction, and severe dizziness and ataxia were present for approximately 48 hours before spontaneous resolution. Based upon his symptoms we estimate an 11-fold increase in plasma concentrations of lidocaine. DISCUSSION: Transdermal delivery of medication affords ease of application, ability to provide continuous pharmacological therapy, and ability to supply medication to the affected site. Due to the superficial application of these medications, external environmental factors may influence the pharmacokinetics of drug delivery. This case highlights the complications that can arise from the combination of heating pads and transdermal pain therapies. PMID- 20026956 TI - Acupuncture for lower back pain: a review. AB - OBJECTIVE: We briefly discuss the history of acupuncture and its postulated mechanisms of action, but our primary objective is to discuss the evidence for acupuncture's efficacy in low back pain as well as approaches of newer study protocols to define more clearly the true usefulness of this alternative modality in low back pain. METHODS: Pubmed online search of all articles and other literature in the past 50 years related to acupuncture efficacy in low back pain, including case reports, randomized controlled trials and meta-analyses. RESULTS: Lower back pain and its associated incapacitating sequelae constitute an important healthcare and socioeconomic problem. There have been multiple, generally poor quality studies on the efficacy of acupuncture for this multi factorial pain condition. Although newer studies seem to show promise, effectiveness has not been clearly demonstrated. CONCLUSIONS: There is a paucity of high-quality research assessing efficacy of acupuncture in the management of LBP. Nonetheless, it continues to play a significant role in our clinical practice, not as a sole therapeutic modality but rather as an adjunct to a multidisciplinary integrative approach of LBP management. Most of the published articles about acupuncture in the biomedical literature consist of case reports, case series, or intervention studies with designs inadequate to assess its efficacy. Thus it is imperative that further research be performed, both preclinical to help elucidate the mechanisms underlying acupuncture, and clinical to justify its clinical application. PMID- 20026957 TI - The concept of addiction in law and regulatory policy related to pain management: a critical review. AB - OBJECTIVE: To present a critical appraisal of the present definitions of addiction-related terminology that appear in US laws and regulatory policies that apply to the prescription of controlled substances for pain management. METHODS: To establish an appropriate context for existing policy definitions, a historical review was conducted of reports from the World Health Organization expert committees on addiction-related concepts, beginning in 1950. In addition, current World Health Organization and American Psychiatric Association diagnostic classification nomenclature were examined. Results from recent criteria-based evaluations of federal and state laws and regulatory policies containing addiction-related terminology also were referenced. RESULTS: Numerous examples are provided to clarify how inaccurate understandings of the nature of addiction, which can be corroborated by archaic definitions in some states' laws, can impact treatment decisions and patient care. Finally, this article discusses terminological and treatment implications of such concepts as "risk mitigation" and "responsible prescribing", which are goals currently emphasized in the pain management field as principal means to reduce addiction to or abuse of prescription opioid medications. DISCUSSION: Although notable improvement has been achieved, policy content in some states has not kept pace with advancements in medical and scientific knowledge about the interface between pain management and addictive disease. Effective translation of addiction-related concepts into clinical practice remains an important objective for promoting public health related to treating pain and reducing non-medical use of opioids. PMID- 20026958 TI - Balance, change, and responsible prescribing: clinicians need updated regulatory policies and the tools to evaluate patients and meet clinical and regulatory obligations! PMID- 20026959 TI - Multimodal, multiclass, multidisciplinary therapy: the key to better analgesia in the 21st century? PMID- 20026960 TI - Delta and kappa opioid receptors as suitable drug targets for pain. AB - Similar to mu opioid receptors, kappa and delta opioid receptors reside in the periphery, the dorsal root ganglion, the spinal cord, and in supraspinal regions associated with pain modulation. Both delta and kappa opioid agonists have been shown to activate pain inhibitory pathways in the central nervous system. Yet, currently there are only a few pharmacologic agents that target kappa receptors, and none that target delta receptors. Spurred by the need for an efficacious analgesic without the unwanted side effects associated with the typical clinical profile of mu opioid agonists, new research has provided insight into why the development of effective kappa and delta opioid receptor agonists has remained elusive thus far, and importantly, how these obstacles may be overcome. For example, for delta opioid agonists to be effective, a state of inflammation may be required as this induces delta opioid receptors to migrate to the surface of neuronal cells and thereby become accessible to delta opioid agonists. Studies have shown that delta opioid agonists can provide relief of inflammatory pain and malignant bone pain. Meanwhile, peripherally restricted kappa opioid agonists have been developed to target kappa opioid receptors located on visceral and somatic afferent nerves for relief of inflammatory, visceral, and neuropathic chronic pain. The recently shown efficacy of these analgesics combined with a possible lower abuse potential and side effect burden than mu opioid receptor agonists makes delta and peripherally restricted kappa opioid receptor agonists promising targets for treating pain. PMID- 20026961 TI - Clinical implications of opioid pharmacogenetics. AB - The peer-reviewed literature yields a plethora of examples of variability in patient's responses to medications. The rapidly progressing field of pharmacogenetics offers insight into the variation in responses observed clinically, and in particular for the variability observed among patients administered mu opioid analgesics. Genetic variation leads to interperson variability in drug absorption, distribution, metabolism, and excretion, processes that have an important impact on the observed efficacy and toxicity of a drug. In particular, single-nucleotide polymorphisms (SNPs) in the gene encoding the mu opioid receptor have been linked to the variability in responses to opioids, whereas SNPs within metabolic enzymes that process and eliminate opioids and their metabolites also have an important effect on an individual's response to opioid medications as do SNPs that affect the bioavailability of opioids. In current clinical practice, given the best available evidence, to optimize pain medications each patient is, in effect, given their own analgesic trial. In the near future, pharmacogenetic approaches may be implemented to best predict which medicine from the outset may be most appropriate for an individual the therapy with the most sustained efficacy and the best side effect profile. In the meantime, pharmacogenetic studies on mu opioid analgesics have provided a molecular foundation supporting opioid rotation in cases in which opioid therapy loses efficacy or becomes associated with intolerable side effects. As more pharmacogenetic research links specific polymorphisms to the pharmacologic effects of specific opioid analgesics, clinicians will continue to improve their understanding of how to prescribe these medications more effectively. PMID- 20026962 TI - Molecular insights into mu opioid pharmacology: From the clinic to the bench. AB - Most of the opioids used in clinical practice exert their effects through mu opioid receptors. Yet, subtle but important pharmacological differences have been observed among the mu opioids. Their potency, effectiveness, and adverse effects can vary unpredictably among patients. These clinical differences among the mu opioids strongly argue against a single receptor mediating their actions. The cloning of the mu opioid receptor has greatly enhanced our understanding of the complexity of this system and has provided possible mechanisms to explain these observations. A single mu opioid receptor gene has been identified, but we now know that it generates a multitude of different mu opioid receptor subtypes through a mechanism commonly used to enhance protein diversity, alternative splicing. Early studies identified a number of splice variants involving the tip of the C-terminus. This region of the receptor is far away from the binding pocket, explaining why these variants still exhibit the same selectivity for mu opioids. However, the differences in structure at the C-terminus influence the activation patterns of the mu opioids. In addition, a second series of variants has been isolated that involves alternative splicing at the N-terminus. Together, these sets of mu opioid receptor splice variants may help explain the clinical variability of the mu drugs among patients and provide insights into why it is so important to individualize therapy for every patient in pain. PMID- 20026963 TI - The prognostic value of F-18 fluorodeoxyglucose bone marrow uptake in patients with recent diagnosis of multiple myeloma: a comparative study with Tc-99m sestamibi. AB - PURPOSE: We assessed the prognostic value of F-18 fluorodeoxyglucose (FDG) uptake in the bone marrow of patients with multiple myeloma (MM) in comparison with Tc 99m methoxy-isobutyl-isonitrile (MIBI). METHODS: The extent and intensity of FDG and MIBI uptake in the bone marrow of 18 patients with a recent diagnosis of MM were assessed by visual score and by calculating the mean SUV (mSUV) for FDG and the femora/thigh ratio (TG/BKG, [Target/Background ratio]) for MIBI images. These parameters were correlated with clinical indexes of disease using hemoglobin and beta-2-microglobulin levels and plasma cell infiltrate (PCI) percentage. The mean values of the visual score, mSUV, and TG/BKG levels were compared in patients deceased after a relatively short follow-up (n = 9; group A) and in patients with a longer survival or were alive at the end of the study (n = 9; group B). RESULTS: Significant correlations of mSUV and TG/BKG values with PCI percentages and beta-2-microglobulin were found (P < 0.05). The extent of FDG and MIBI bone marrow uptake was greater in patients of group A (P < 0.01). Higher values of mSUV (P < 0.01) and TG/BKG (P < 0.05) were also observed in patients of group A. These results were consistent with the differences (not statistically significant) in hemoglobin, albumin, beta-2-microglobulin levels, and PCI percentages observed in the 2 groups. CONCLUSION: Our study demonstrates that an increase of FDG bone marrow uptake may predict a more aggressive disease, as much as MIBI uptake. Therefore, an additional analysis of FDG bone marrow images should be performed in patients undergoing PET studies during the initial staging of MM. PMID- 20026964 TI - Detection of residual lymph node metastases in high-risk papillary thyroid cancer patients receiving adjuvant I-131 therapy: the usefulness of F-18 FDG PET/CT. AB - OBJECTIVE: The aims of this retrospective study were to evaluate the incidence of residual lymph node (LN) metastases in high-risk papillary thyroid cancer (PTC) patients receiving adjuvant I-131 therapy, especially in those without I-131 accumulation, and to evaluate the clinical usefulness of F-18 FDG PET/CT to detect those lesions. METHODS: The 37 PTC patients receiving adjuvant I-131 therapy were retrospectively evaluated. We examined the incidence of residual LN metastasis and compared the accumulation of F-18 FDG and I-131 in those lesions, and compared the serum thyroglobulin (Tg) levels between patients with and without residual LN metastases. RESULT: A total of 33 lesions were diagnosed as residual LN metastases in 9 patients. FDG accumulated in all of the lesions, but 19 lesions (57.6%) of them had no I-131 accumulation. The SUVmax was significantly higher in lesions without I-131 accumulation than in lesions with I 131 accumulation (6.6 +/- 2.8 vs. 4.2 +/- 1.8; P = 0.007). The serum Tg levels were significantly higher in patients with residual LN metastases than in patients without it (709.9 +/- 1470.8 vs. 25.6 +/- 37.1 ng/mL; P = 0.005). The incidence of residual LN metastasis was significantly higher in patients with a detectable serum Tg level than in patients without it (35.0% vs. 0%, P = 0.03). CONCLUSION: These results indicate that the residual LN metastasis was relatively common in high-risk PTC patients receiving adjuvant I-131 therapy whose serum Tg levels remained detectable, and those lesions often had no I-131 accumulation. FDG-PET/CT would be recommended for PTC patients with detectable serum Tg levels to detect residual LN metastases. PMID- 20026965 TI - Clinical impact of SPECT/CT with In-111 biotin on the management of patients with suspected spine infection. AB - PURPOSE: Early identification and localization of spine infection is necessary for adequate therapeutic strategy. To localize the precise site of infection we evaluated In-111 Biotin SPECT/CT versus planar and SPECT imaging. METHODS: Seventy-two consecutive patients were enrolled and underwent SPECT/CT and planar imaging 2 to 4 hours post i.v. injection of In-111 Biotin. Final diagnosis was based on bacterial cultures and/or clinical/imaging follow-up for at least 1 year. We evaluated the diagnostic performance of planar, SPECT, and SPECT/CT In 111 Biotin scintigraphy. RESULTS: In-111 Biotin SPECT/CT and SPECT showed similar values of sensitivity (93.5% vs. 92.1%) and the same specificity (92.3%), planar imaging showed 80.4% of sensitivity and 69.2% of specificity. In 16 patients SPECT/CT correctly localized the infection site (bone, soft tissue, or both bone and soft tissue). CONCLUSIONS: SPECT/CT enhances the impact of In-111 Biotin scintigraphy on the clinical management of patients, allowing the exact site of infection to be localized to select the appropriate therapy. PMID- 20026966 TI - F-18 FDG PET/CT findings in two patients with hepatic angiomyolipoma with and without intratumoral hemorrhage. AB - Two female patients aged 62 and 74 years underwent F-18 FDG PET/CT for evaluation of a large hepatic tumor. Subsequently, both patients underwent a partial hepatectomy which histopathologically revealed benign hepatic angiomyolipomas (AMLs). One patient with AML with a massive intratumoral hemorrhage presented with multiple foci demonstrating an increased FDG uptake along the margin of the cold defects that corresponded to low density areas within the tumor on contrast enhanced CT, whereas the other patient with AML without any hemorrhage presented with a low FDG uptake in the tumor. This report demonstrates that benign hepatic AMLs may demonstrate increased FDG uptake if there is hemorrhage and a related inflammatory response. PMID- 20026967 TI - Interesting image. F-18 fluorodeoxyglocose positron emission tomography/computed tomography in the diagnosis of chronic myopathic sarcoidosis. PMID- 20026968 TI - Interesting image. FDG-PET/CT in primary large B-cell lymphoma of the hard palate. PMID- 20026969 TI - Interesting image. Situs inversus totalis in newly diagnosed lymphoma: additional value of hybrid imaging. PMID- 20026970 TI - Interesting image. Maffucci syndrome with extraosseous chondrosarcoma imaged with F-18 FDG PET-CT. PMID- 20026971 TI - Interesting image. Diagnosis of acute osteomyelitis in previously irradiated bone: which test to use? PMID- 20026972 TI - Interesting image. Incidental diagnosis of prostate cancer by F-18 FDG PET/CT. PMID- 20026973 TI - Interesting image. Tc-99m sestamibi SPECT/CT as a new tool for monitoring perfusion and viability of buried perforator based free flaps in breast reconstruction after breast cancer. PMID- 20026974 TI - Interesting image. Extrapulmonary small cell carcinoma in prostate: detection with F-18 FDG PET/CT. PMID- 20026975 TI - Interesting image. Pancreatic neuroendocrine tumor with involvement of the inferior mesenteric vein diagnosed by Ga-68 DOTA-TATE PET/CT. PMID- 20026976 TI - Interesting image. Correlative imaging with FDG PET-CT and MRI of an unusual recurrence of urothelial cancer in an ileal conduit. PMID- 20026977 TI - Interesting image. Utility of FDG metabolism to differentiate synchronous metastatic liver lesions from synchronous colon cancer: nonmucinous versus mucinous adenocarcinoma. PMID- 20026978 TI - Interesting image. Serial PET changes of extraocular muscle hypermetabolism in sporadic fatal insomnia. PMID- 20026979 TI - Interesting image. PET imaging of a butterfly glioblastoma. PMID- 20026980 TI - Interesting image. I-123 iomazenil SPECT imaging in a patient with hemispheric brain damage due to cerebral ischemia during carotid endarterectomy. PMID- 20026981 TI - Interesting image. Two primary colon cancers shown on FDG PET/CT performed to evaluate possible lung metastases from bladder cancer. PMID- 20026982 TI - Interesting image. Ameboma mimicking lung cancer on FDG PET/CT. PMID- 20026983 TI - Current readings in nuclear medicine. PMID- 20026987 TI - Current world literature. PMID- 20026985 TI - Evidence-based medicine and health care reform legislation. PMID- 20026989 TI - Current world literature. PMID- 20026992 TI - Achieving cost control, care coordination, and quality improvement through incremental payment system reform. AB - The healthcare reform goal of increasing eligibility and coverage cannot be realized without simultaneously achieving control over healthcare costs. The reform of existing payment systems can provide the financial incentive for providers to deliver care in a more coordinated and efficient manner with minimal changes to existing payer and provider infrastructure. Pay for performance, best practice pricing, price discounting, alignment of incentives, the medical home, payment by episodes, and provider performance reports are a set of payment reforms that can result in lower costs, better coordination of care, improved quality of care, and increased consumer involvement. These reforms can produce immediate Medicare annual savings of $10 billion and create the framework for future savings by establishing financial incentives for long-term provider behavior changes that can lead to lower costs. PMID- 20026993 TI - Response to Achieving cost control, care coordination, and quality improvement through incremental payment system reform. PMID- 20026994 TI - Perspective: the potential for incremental payment system reform. PMID- 20026995 TI - Guiding principles for payment system reform: commentary on "Achieving cost control, care coordination, and quality improvement through incremental payment system reform". AB - The focus of healthcare payment reform must be on the "value equation"--improving the quality of care while reducing or controlling costs. Adhering to key principles related to value will yield successful payment transformation and result in more coordinated care and improved patient outcomes. Actionable data and information for setting goals and measuring progress is an essential element. PMID- 20026996 TI - Payment reform to finance a medical home: comment on "Achieving cost control, care coordination, and quality improvement through incremental payment system reform". AB - This commentary on R. F. Averill et al. (2010) addresses their idea of risk and quality adjusting fee-for-service payments to primary care physicians in order to improve the efficiency of primary care and take a step toward financing a "medical home"for patients. I show how their idea can create incentives for efficient practice styles. Pairing this with an active beneficiary choice of primary care physician with an enrollment fee would make the idea easier to implement and provide an incentive and the financing for elements of service not covered by procedure-based fees. PMID- 20026997 TI - Achieving cost control, care coordination, and quality improvement in the Medicaid program. AB - As Medicaid spending rises quickly, controlling cost while coordinating care and improving quality is paramount. Payment method reforms should reward providers that deliver beneficial care but not care of marginal value. This article draws lessons from previous payment reforms and analyzes the potential benefits for Medicaid of 7 specific proposals. The most promising are paying for quality (especially rewarding hospitals that reduce rates of potentially preventable complications and readmissions) and moving away from fee-for-service payment for "medical home" providers. To achieve reform, the article recommends a process characterized by persistence, attention to outliers, consultation, and timing. PMID- 20026998 TI - Payment system reform for Medicaid in New York: comment on "Achieving cost control, care coordination, and quality improvement through incremental payment system reform". PMID- 20026999 TI - Getting from here to there in payment reform: necessary practices and policies. PMID- 20027000 TI - Comment on "Achieving cost control, care coordination, and quality improvement through incremental payment system reform". PMID- 20027001 TI - Building a national payment system: comment on "Achieving cost control, care coordination, and quality improvement through incremental payment system reform". PMID- 20027002 TI - Maryland's approach in enhancing effectiveness and efficiency in healthcare delivery: comment on "Achieving cost control, care coordination, and quality improvement through incremental payment system reform". PMID- 20027004 TI - Building the affordable medical home. AB - Medical homes and accountable care organizations have become the great hope for many health industry stakeholders and policy makers seeking to reconstitute fragmented healthcare delivery within a coordinated cost-conscious framework. The need to increase coordination has been evident to analysts for many years and, in the past, has resulted in developments such as capitation and disease management. The hope of these reforms is that through changes in risk/reward to providers or by layering in external programs to improve patient coordination the system will improve patient outcomes and reduce total cost. This article describes a pilot project to create a medical home that is built upon changing existing payment incentives but tying these incentives directly to the outcomes of care experienced by patients. It is at variance with most proposals to create medical homes in that it is self-funded, allowing rapid scaling, and is largely agnostic toward the precise practice pattern employed. Put succinctly, it encourages practice patterns that improve patient outcomes without dictating what they should be. PMID- 20027005 TI - The Accountable Care Organization (ACO) model: building blocks for success. AB - The Accountable Care Organization (ACO) model has received significant attention among policymakers and leaders in the healthcare community in the context of the ongoing debate over health reform, not only because of the unsustainable path on which the country now finds itself but also because it directly focuses on what must be a key goal of the healthcare system: higher value. The model offers a promising approach for achieving this goal. This article provides an overview of the ACO model and its role in the current policy context, highlights the key elements that will be common to all ACOs, and provides details of several challenges that may arise throughout the implementation process, including a host of technical, legal, and operational challenges. These challenges range from issues such as the organizational form and management of the ACO to analytic challenges such as the calculation of spending benchmarks and the selection of quality measures. PMID- 20027006 TI - What if ...: a patient makes a sexual advance toward you? PMID- 20027008 TI - Small intestinal bacterial overgrowth in gastroparesis: are there any predictors? AB - BACKGROUND: There is some degree of overlap in the symptomatic spectrum of patients with gastroparesis and small intestinal bacterial overgrowth (SIBO) and some of the etiologies for gastroparesis, such as diabetes mellitus and vagotomy are known to predispose to SIBO. The aims of our study were to measure the prevalence of SIBO in a cohort of gastroparetic patients with prominence of abdominal pain and bloating and try to identify predictors with regard to demographics, concurrent use of medications such as prokinetics, proton pump inhibitors, and opiate analgesics, and predominant bowel movement abnormality. METHODS: Glucose breath testing (GBT) for SIBO was performed in 50 patients (41 females) with gastroparesis. Demographic data, medication profiles, baseline and peak measurements of hydrogen or methane gas on the GBT, and results of the most recent gastric emptying scintigraphy test were recorded. RESULTS: Thirty of fifty (60%) patients had a positive GBT for SIBO on the basis of hydrogen (63%), methane (27%), or both criteria (10%). SIBO was more likely (P=0.001) in patients with gastroparetic symptoms of greater duration (mean 5 y; 95% CI: 4-6 y). No significant differences were noted in both groups with regard to age, sex, or etiology of gastroparesis. Gastric emptying was similar in the SIBO and non-SIBO group (P>0.05). After adjusting for tegaserod and opiate analgesic use, 14/23 (61%) had a positive GBT. CONCLUSIONS: SIBO is very common in gastroparetics with predominance of abdominal pain and bloating, especially those with a longer duration of gastroparesis. Awareness of SIBO in the setting of gastroparesis will facilitate separation of the 2 entities and allow appropriate therapies to be instituted. PMID- 20027009 TI - Celiac disease: prevention and treatment. AB - Celiac disease is a common multisystemic disorder that may be diagnosed at any age. Owing to the increasing prevalence, prevention strategies, including encouragement of breastfeeding and introduction of small amounts of gluten whereas still on breastfeeding at fourth to sixth month of age, become more important than before. For avoiding complications of celiac disease, a strict gluten-free diet is essential. A lifelong gluten-free diet may bring about difficulties as avoiding gluten completely is problematic owing to the contamination with gluten of presumably gluten free foods. New therapeutic approaches include enzyme supplementation, correction of the intestinal barrier defect against gluten entry, blocking of gliadin presentation by human leukocyte antigen blockers and tissue transglutaminase inhibitors. PMID- 20027010 TI - Efficacy of tricyclic antidepressant therapy in adults with cyclic vomiting syndrome: a two-year follow-up study. AB - AIM: To investigate the effects of prolonged tricyclic antidepressants (TCAs) therapy on the frequency and duration of episodes of cyclic vomiting syndrome (CVS) in adults, and the global assessment of clinical improvement and the number of emergency department (ED) visits and/or hospitalizations. METHOD: An open labeled study was conducted in adult CVS patients treated with a TCA and followed for up to 2 years. Demographic data, TCA dosage, duration and frequency of CVS episodes, ED visits, and hospitalizations at baseline and during TCA therapy were recorded, and patients were monitored for any adverse events. RESULTS: Forty-six patients initially met the inclusion criteria and 41 patients (22 male) with mean age of 35 years (range: 18 to 63 y) on TCA were able to be followed for 1 year and 23 were evaluated for 2 years. Mean age of onset of CVS symptoms was 26 years (range: 10 to 52 y) and mean age for making the diagnosis of CVS was 32 years (range: 15 to 63 y). The frequency and duration of an episode and ED visits related to CVS were all significantly reduced after both the first and second year of TCA therapy (P<0.05). Eighty-eight percent reported an improved clinical status by subjective global assessment. Mild side effects were reported in 34% not necessitating stoppage of the TCA. CONCLUSIONS: Long-term TCA therapy significantly reduces the frequency and duration of CVS episodes, ED visits, and hospitalizations (P<0.05), and improves overall clinical well-being providing evidence that they are effective therapy for adult CVS. PMID- 20027011 TI - The prognostic value of NSE and S100B from serum and cerebrospinal fluid in patients with spontaneous subarachnoid hemorrhage. AB - Neuron-specific enolase (NSE) and S100B protein have been shown to be increased in cerebrospinal fluid (CSF) and serum of patients suffering from subarachnoid hemorrhage. This study was designed to evaluate the accuracy of NSE and S100B from CSF and serum for the prognosis of outcome and the detection of cerebral infarction, vasospasm and intracranial hypertension. In 55 patients with spontaneous subarachnoid hemorrhage and requiring external ventricular drainage the concentrations of NSE and S100B were determined daily from the serum and the CSF from admission until day 8. At ICU discharge patients' outcome was assessed by the Glasgow outcome scale and occurrence of cerebral infarction, vasospasm and intracranial hypertension were registered. Mean and peak values of each parameter for each patient were calculated. For accuracy assessment receiver operating characteristics were used. Bad outcome (Glasgow outcome scale 1 to 3) was found in 33 patients. Cerebral infarction, vasospasm, and intracranial hypertension were found in 31 (56%), 34 (62%), and 36 (65%) patients. Mean and peak values of NSE CSF (P<0.001), S100B CSF (P<0.001), and S100B serum (P<0.001) but not of NSE serum provided the ability to distinguish between patients with good and bad outcome. The accuracy of NSE CSF and S100B CSF did not differ significantly from that of S100B serum. NSE CSF (P<0.001), S100B CSF (P<0.001), and S100B serum (P<0.001) allowed the detection of cerebral infarction and intracranial hypertension. Cerebral vasospasm was detected by none of the parameters. In conclusion, NSE CSF, S100B CSF, and S100B serum provide similar prognostic values for outcome, intracranial hypertension and cerebral infarction. Significantly lower accuracy was found for NSE serum. PMID- 20027012 TI - Retrospective analysis of the risk factors and pathogens associated with early onset ventilator-associated pneumonia in surgical-ICU head-trauma patients. AB - BACKGROUND: Early-onset ventilator associated pneumonia (EOVAP) are frequent in head-trauma patients, but specific risk factors are poorly studied in this population. METHODS: We conducted a retrospective cohort study in a surgical intensive care unit. Consecutive severe head-trauma patients admitted from January 2000 to December 2002 were studied. Microorganisms, and risks factors for EOVAP were analyzed. RESULTS: During the 3-year period, 161 patients were studied; 21.1% of them developed an EOVAP. On univariate analysis 6 variables were associated with EOVAP: early enteral feeding, barbiturate use, immunosuppression, mean Simplified Acute Physiology Score 2, acute respiratory distress syndrome, and initial neurosurgery procedures. On multivariate analysis, enteral feeding >2000 Kcal before day 5 [odds ratio (OR): 0.33, 95% confidence interval (CI): 0.21-0.85] and initial neurosurgical procedure (OR: 0.36, 95% CI: 0.15-0.89) remained protective factors for EOVAP, whereas immunosuppression (OR: 7.15, 95% CI: 1.66-30.73) and barbiturate use (OR: 2.68, 95% CI: 1.06-6.80) remained risk factors for EOVAP. EOVAP was also significantly associated with a longer duration of mechanical ventilation (14.0 vs. 11.0 d, P=0.024), and a longer sedation duration (8.3 vs. 5.8 d P=0.005). Methicillin-susceptible Staphylococcus aureus was the most common pathogen involved in EOVAP (46%). CONCLUSIONS: We demonstrate for the first time that early enteral feeding is a protective factor for EOVAP, and this result could have clinical implications for the prevention of EOVAP after traumatic brain injury. This study also confirms that barbiturate use is an important risk factor of EOVAP whereas Methicillin susceptible S. aureus was found to be the main pathogen involved in EOVAP. PMID- 20027013 TI - Cerebral aneurysm surgery in a patient with Brugada syndrome: anesthetic implications and perioperative management. PMID- 20027014 TI - The timeline of our lives. PMID- 20027016 TI - Your role in preventing identity theft. PMID- 20027017 TI - Is PSA necessary after prostatectomy? PMID- 20027018 TI - E-learning resources for acute care nurse practitioners. PMID- 20027019 TI - Acquired fecal incontinence in community-dwelling adults. PMID- 20027021 TI - 22nd Annual Legislative Update: Regulatory and legislative successes for APNs. PMID- 20027023 TI - A new treatment option for type 2 diabetes. PMID- 20027026 TI - The truth about inherited thrombophilias and pregnancy. PMID- 20027027 TI - Inherited thrombophilia polymorphisms and pregnancy outcomes in nulliparous women. AB - OBJECTIVE: To estimate the association between five commonly inherited thrombophilia polymorphisms and adverse pregnancy outcomes in women who had no prior history of adverse pregnancy outcomes or personal or family history of venous thromboembolism. METHODS: Healthy nulliparous women (n=2,034) were recruited to this prospective cohort study before 22 weeks of gestation. Genotyping for factor V Leiden, prothrombin gene mutation, methylenetetrahydrofolate reductase enzyme (MTHFR) C677T, MTHFR A1298C, and thrombomodulin polymorphism was performed. Clinicians caring for women were blinded to the results of thrombophilia tests. The primary composite outcome was the development of severe preeclampsia, fetal growth restriction, placental abruption, stillbirth, or neonatal death. RESULTS: Complete molecular results and pregnancy outcome data were available in 1,707 women. These complications were experienced by 136 women (8.0%). Multivariable logistic regression demonstrated two statistically significant findings. Women who carried the prothrombin gene mutation had an odds ratio of 3.58 (95% confidence interval [CI] 1.20-10.61, P=.02) for the development of the composite primary outcome. Homozygous carriers of the MTHFR 1298 polymorphism had an odds ratio of 0.26 (95% CI 0.08-0.86, P=.03). None of the other polymorphisms studied showed a significant association with the development of the primary outcome in this cohort of women. CONCLUSION: Prothrombin gene mutation confers an increased risk for the development of adverse pregnancy outcomes in otherwise asymptomatic, nulliparous women, whereas homozygosity for MTHFR 1298 may protect against these complications. The majority of asymptomatic women who carry an inherited thrombophilia polymorphism have a successful pregnancy outcome. LEVEL OF EVIDENCE: II. PMID- 20027028 TI - Prothrombin gene G20210A mutation and obstetric complications. AB - OBJECTIVE: To estimate whether maternal carriage of the prothrombin gene G20210A mutation is associated with pregnancy loss, preeclampsia, placental abruption, or small for gestational age (SGA) neonates in a low-risk, prospective cohort. METHODS: This was a secondary analysis of the Eunice Kennedy Shriver National Institute of Child Health and Human Development factor V Leiden study, a multicenter, prospective, observational cohort of 5,188 unselected singleton gestations. A total of 4,167 first-trimester samples were available for analysis and were tested for the prothrombin G20210A mutation. Obstetric complications were compared between women with and without the prothrombin G20210A mutation by univariable and multivariable analysis. RESULTS: A total of 157 (3.8%) women had the prothrombin gene mutation (156 heterozygous and one homozygous). Carriers of the prothrombin G20210A mutation had similar rates of pregnancy loss, preeclampsia, SGA neonates, and abruption compared with noncarriers. Results were similar in a multivariable analysis controlling for age, race, prior pregnancy loss, prior SGA neonates, and family history of thromboembolism. Three thromboembolic events occurred in women testing negative for the mutation. CONCLUSION: There was no association between the prothrombin G20210A mutation and pregnancy loss, preeclampsia, abruption, or SGA neonates in a low-risk, prospective cohort. These data raise questions about the practice of screening women without a history of thrombosis or adverse pregnancy outcomes for this mutation. LEVEL OF EVIDENCE: II. PMID- 20027029 TI - Effects of magnesium sulfate on preterm fetal cerebral blood flow using Doppler analysis: a randomized controlled trial. AB - OBJECTIVE: To estimate the effects of maternal administration of magnesium sulfate on the fetal middle cerebral artery using Doppler. METHODS: This is a single-center ancillary study done in conjunction with a randomized, double masked, placebo- controlled trial in 20 centers. Women at imminent risk of preterm delivery (n=2,241) were allocated randomly to receive magnesium sulfate or placebo. Doppler measurements of the fetal middle cerebral artery were obtained before blinded study-drug administration and subsequently at 1-, 2-, 3-, and 4-hour intervals. Parameters studied included time-average velocity, peak systolic velocity, vessel diameter, heart rate, and calculated blood volume flow. A random-effects model with repeated-measures design was used for analysis. RESULTS: A total of 38 fetuses were studied: 18 received magnesium sulfate and 20 received placebo. Peak systolic velocity was significantly related to gestational age (P<.001). There were no differences between the study groups for middle cerebral artery peak systolic velocity, average velocity, vessel diameter, or calculated volume flow. However, fetal heart rate significantly decreased after treatment with magnesium sulfate. CONCLUSION: Magnesium sulfate had no significant effects on fetal cerebral blood flow analyzed using Doppler. The only parameter in the fetal cerebral circulation significantly modified by magnesium sulfate was the heart rate. The significance of this heart-rate change, vis-a-vis the neuroprotective effects of magnesium sulfate, is unknown. CLINICAL TRIAL REGISTRATION: ClinicalTrials.gov, www.clinicaltrials.gov, NCT00014989. LEVEL OF EVIDENCE: I. PMID- 20027030 TI - Paracervical block efficacy in office hysteroscopic sterilization: a randomized controlled trial. AB - OBJECTIVE: To estimate the efficacy of paracervical block compared with saline for pain relief during office hysteroscopic sterilization. METHODS: This study was a randomized, placebo-controlled study of women desiring hysteroscopic sterilization. A paracervical block of 1% lidocaine or normal saline was administered before office hysteroscopic sterilization. Patients and investigators were blinded to assignments. A pre hoc power analysis determined that 40 women would be required per arm to detect a difference of 0.9 cm on a visual analog scale. Pain was recorded on a visual analog scale at multiple procedure time points. Individualized standardized pain scores were constructed by weighted reporting of objective and subjective sensation. RESULTS: A total of 103 consecutive women were eligible, and 80 women were randomized, with 40 per group. Thirty-seven (93%) in each group had successful placement. The lidocaine group showed significantly lower pain scores for tenaculum placement (mean+/ standard deviation: 0.97+/-1.28 compared with 3.00+/-2.41, P<.001) traversing the external cervical os (1.46+/-1.71 compared with 3.77+/-2.68, P<.001) and internal os (1.79+/-2.11 compared with 4.10+/-2.77, P<.001). There was no significant observed difference with device placement in tubal ostium (3.15+/-2.69 compared with 3.74+/-2.73, P=.33). Multivariable linear regression analysis demonstrated a relationship of pain to procedural time (P=.047) and to group assignment (P<.01). CONCLUSION: Paracervical block with 1% lidocaine provides effective pain relief for cervical manipulations during office hysteroscopic sterilization, but does not reduce the pain associated with upper uterine/tubal manipulation when placing the devices. CLINICAL TRIAL REGISTRATION: ClinicalTrials.gov, www.clinicaltrials.gov, NCT00811187. LEVEL OF EVIDENCE: I. PMID- 20027031 TI - Predictors of higher bone mineral density loss and use of depot medroxyprogesterone acetate. AB - OBJECTIVE: To identify possible predictive factors of higher bone loss, defined as at least 5%, at the spine or femoral neck, over time in depot medroxyprogesterone acetate (DMPA) users. METHODS: Bone mineral density (BMD) was measured at the lumbar spine and femoral neck every 6 months in 240 white, African-American, and Hispanic women using DMPA. For the purpose of analysis, an arbitrary value of at least 5% BMD loss from the baseline value after 24 months of DMPA use at either the lumbar spine or the femoral neck was considered as higher BMD loss. Logistic regression analysis was then used to examine factors predictive of at least 5% BMD loss at either site. RESULTS: Of the initial 240 DMPA users, 95 completed 24 months of follow-up. Forty-five of the 95 DMPA users (47.4%) had at least 5% BMD loss at the lumbar spine or femoral neck by 24 months. Multivariable logistic regression model showed that at least 5% BMD loss was associated with current smoking (adjusted odds ratio [OR] 3.88, 95% confidence interval [CI] 1.26-11.96), calcium intake (in 100 mg) (OR 0.81, 95% CI 0.65-0.99), and parity (OR 0.49, 95% CI 0.29-0.82). Age, race or ethnicity, previous contraceptive use, and body mass index were not associated with higher BMD loss. CONCLUSION: The risk of higher BMD loss associated with DMPA use may be reduced by quitting smoking and increasing calcium intake. Having had a child is also protective. LEVEL OF EVIDENCE: II. PMID- 20027032 TI - Lactation and maternal measures of subclinical cardiovascular disease. AB - OBJECTIVE: To examine the relationship between lactation and subclinical cardiovascular disease in a population free of clinical cardiovascular disease. METHODS: We conducted a cross-sectional analysis of 297 women who reported at least one live birth on enrollment in the Study of Women Across the Nation-Heart Study. Participants were mothers aged 45-58 years who were free of clinical cardiovascular disease. History of lactation was self-reported. Electron beam tomography was used to assess coronary and aortic calcification. B-mode ultrasonography was used to assess carotid adventitial diameter, intima-media thickness, and carotid plaque. Multivariable linear and logistic regression models were used to estimate whether lactation was independently associated with markers of subclinical cardiovascular disease. RESULTS: In unadjusted models, compared with mothers who had breastfed all of their children for at least 3 months, mothers who had not breastfed were more likely to have coronary artery calcification (17% compared with 32%), aortic calcification (17% compared with 39%), carotid plaque (10% compared with 18%), and larger carotid adventitial diameters (mean+/-standard deviation 6.63+/-0.59 compared with 6.87+/-0.60 mm). After adjusting for measures of socioeconomic status and lifestyle and family history variables, mothers who had not breastfed remained more likely to have aortic calcification (odds ratio [OR] 3.85, 95% confidence interval [CI] 1.47 10.00) and coronary artery calcification (OR 2.78, 95% CI 1.05-7.14) than mothers who had consistently breastfed. After further adjustment for body mass index and traditional risk factors for cardiovascular disease, mothers who had not breastfed remained more likely to have aortic calcification than mothers who had consistently breastfed (OR 5.26, 95% CI 1.47-20.00). CONCLUSION: Mothers who do not breastfeed their infants seem to be at increased risk of vascular changes associated with future cardiovascular disease. LEVEL OF EVIDENCE: II. PMID- 20027033 TI - High poly(adenosine diphosphate-ribose) polymerase expression and poor survival in advanced-stage serous ovarian cancer. AB - OBJECTIVE: To estimate the range of poly(adenosine diphosphate [ADP]-ribose) polymerase expression in serous ovarian cancers and to determine whether expression is associated with response to therapy and outcome. METHODS: Immunostaining for poly(ADP-ribose) polymerase was performed in 186 paraffin embedded, serous ovarian cancers. Nuclear poly(ADP-ribose) polymerase expression was quantified using a scoring system that assesses both staining intensity and percentage of cells staining. Kaplan-Meier analysis was performed to evaluate the relationship between poly(ADP-ribose) polymerase expression and overall survival. RESULTS: High poly(ADP-ribose) polymerase expression was present in 54% of serous cancers but was not associated with stage or grade. There was no difference in the rate of complete clinical response to primary chemotherapy between cases with low poly(ADP-ribose) polymerase expression (70%) compared with those with high poly(ADP-ribose) polymerase expression (71%). However, high poly(ADP-ribose) polymerase expression was associated with significantly worse median overall survival (36 compared with 43 months, P=.04, hazard ratio 0.71). CONCLUSION: Expression of poly(ADP-ribose) polymerase in ovarian cancers is heterogeneous, and high expression in serous ovarian cancers is associated with worse overall survival. These data suggest that evaluation of poly(ADP-ribose) polymerase expression in the primary cancer could potentially allow selective use of poly(ADP-ribose) polymerase inhibitors in patients most likely to respond. LEVEL OF EVIDENCE: III. PMID- 20027034 TI - Metformin compared with glyburide in gestational diabetes: a randomized controlled trial. AB - OBJECTIVE: To compare the efficacy of metformin with glyburide for glycemic control in gestational diabetes. METHODS: Patients with gestational diabetes who did not achieve glycemic control on diet were randomly assigned to metformin (n=75) or glyburide (n=74) as single agents. The primary outcome was glycemic control. Secondary outcomes were drug failure rate and neonatal and obstetric complications. RESULTS: In the patients who achieved adequate glycemic control, the mean fasting and 2-hour postprandial blood glucose levels were not statistically different between the two groups. However, 26 patients in the metformin group (34.7%) and 12 patients in the glyburide group (16.2%) did not achieve adequate glycemic control and required insulin therapy (P=.01). CONCLUSION: In this study, the failure rate of metformin was 2.1 times higher than the failure rate of glyburide when used in the management of gestational diabetes (95% confidence interval 1.2-3.9). CLINICAL TRIAL REGISTRATION: ClinicalTrials.gov, www.clinicaltrials.gov, NCT00965991. LEVEL OF EVIDENCE: I. PMID- 20027036 TI - Effect of predelivery diagnosis in 99 consecutive cases of placenta accreta. AB - OBJECTIVE: To estimate the effects of prenatal diagnosis and delivery planning on outcomes in patients with placenta accreta. METHODS: A review was performed of all patients with pathologically confirmed placenta accreta at the University of California, San Diego Medical Center from January 1990 to April 2008. Cases were divided into those with and without predelivery diagnosis of placenta accreta. Patients with prenatal diagnosis of placenta accreta were scheduled for planned en bloc hysterectomy without removal of the placenta at 34-35 weeks of gestation after betamethasone administration. Maternal and neonatal outcomes were assessed. RESULTS: Ninety-nine women with placenta accreta were identified, of whom 62 were diagnosed before delivery and 37 were diagnosed intrapartum. Comparing women with predelivery diagnosis with those diagnosed at the time of delivery, there were fewer units of packed red blood cells transfused (4.7+/-2.2 compared with 6.9+/ 1.8 units, P=.02) and a lower estimated blood loss (2,344+/-1.7 compared with 2,951+/-1.8 mL, P=.053), although this trend did not reach statistical significance. Comparison of neonatal outcomes demonstrated a higher rate of steroid administration (65% compared with 16%, P0.05) and the postoperative hospital stay was significantly longer (P<0.05) in the OS group compared with the LS group. Scores from the modified Body Image, Hospital Experience, and Photo Series Questionnaires were higher (favorable) in the LS group compared with the OS group (P<0.01, P<0.001, and P<0.001, respectively). CONCLUSIONS: LS is the procedure of choice for most indications of splenectomy, which has comparable or better perioperative outcomes and complication rate as well as the advantages of better body image, cosmesis, and hospital experience compared with OS. Our results presenting improved outcomes with LS will contribute to the widespread application of laparoscopy for splenectomy. PMID- 20027092 TI - The feasibility and effectiveness of a hand-assisted laparoscopic splenectomy for hypersplenism in patients after living-donor liver transplantation. AB - BACKGROUND: A laparoscopic splenectomy in patients who previously underwent living-donor liver transplantation (LDLT) is thought to be technically difficult because of the presence of severe adhesions and splenomegaly. This report documents the efficacy and safety of a hand-assisted laparoscopic splenectomy (HALS) for hypersplenism in patients after LDLT. METHODS: Five patients underwent HALS for hypersplenism after LDLT between 1999 and 2007. The medical records of those patients were retrospectively evaluated. RESULTS: The mean operative time was 237+/-12 minutes. The mean blood loss was 229+/-100 mL and the mean weight of excised spleen was 461+/-46 g. There was no conversion to open surgery. The number of platelets and leukocytes were significantly increased after surgery. No major complications were observed except for a patient who suffered paralytic ileus postoperatively. The mean hospital stay after the operation was 16.7+/-2.5 days. CONCLUSIONS: HALS for patients after LDLT is a feasible and safe procedure. This technique can thus become a standard procedure after LDLT. PMID- 20027093 TI - Postoperative complications in elderly patients with colorectal cancer: comparison of open and laparoscopic surgical procedures. AB - BACKGROUND: Surgery is associated with higher morbidity and mortality rates in elderly patients with colorectal cancer compared with younger patients. The aim of this study was to examine preoperative evaluation for selecting operative procedure in elderly patients with colorectal cancer. METHODS: The study of all patients who underwent open surgery (OS) or laparoscopically assisted surgery (LAS) for colorectal cancer from January 2004 to December 2007 were aged > or =71 years. Preoperative evaluation, operative factors, morbidity, and mortality were analyzed by the Physiological and Operative Severity Score for Enumeration of Mortality and Morbidity (POSSUM) and Prognostic Nutritional Index (PNI). RESULTS: A total of 129 patients were included in this study. Fifty-one patients underwent OS, and LAS was performed on 78 patients. The morbidity rate was 51.3% (40 patients) for the OS group and 23.5% (12 patients) for the LAS group. Three LAS patients (5.9%) subsequently required OS. One LAS patient died postoperatively. There were significant differences in the Operative Severity Score (OSS) in POSSUM and PNI, but not Physiologic Score (PS) in POSSUM, between the two groups. In the OS group, there were significant differences in PS, OSS, and PNI between those with or without complications, whereas in the LAS group, OSS, but not PS or PNI, was significantly lower in those without than in those with complications. CONCLUSIONS: Compared with OS, LAS is associated with a lower incidence of complications in elderly patients with colorectal cancer. The nutritional status correlated with postoperative complications in the OS group. PMID- 20027094 TI - Comparative study on surgical outcomes and operative costs of transumbilical single-port laparoscopic appendectomy versus conventional laparoscopic appendectomy in adult patients. AB - PURPOSE: To reduce the incisional morbidity and improve cosmesis, the authors adopted a single-port laparoscopic appendectomy using a unique "single-port". The authors retrospectively evaluated transumbilical single-port laparoscopic appendectomy (TUSPLA) against conventional laparoscopic appendectomy (CLA) with respect to perioperative outcomes and operative cost in the adult patients. MATERIALS AND METHODS: The authors reviewed the case of laparoscopic appendectomy between April 2008 and January 2009. Demographic data, operating time, pathology of appendix, hospital stay, surgical morbidities, visual analogue scale pain scores, analgesics requirements, and the operative cost were compared. RESULTS: Seventy-two underwent TUSPLA and 108 underwent CLA. There were more females in the TUSPLA group. Surgical outcomes were similar between the TUSPLA and the CLA group. Operative cost was significantly lower in the TUSPLA group compared with the CLA group. CONCLUSIONS: This study showed that TUSPLA was cost saving and produced similar surgical outcomes in selected appendicitis patients compared with CLA. PMID- 20027095 TI - Treatment of incisional hernias adopting an intra-abdominal approach with a new low-density composite prosthetic material: proceed: our preliminary experience on 50 cases. AB - INTRODUCTION: The management of incisional hernias remains a challenge for the general surgeon. Repairing by using prosthetic materials has reduced the relapse rate, but intra-abdominal mesh placement continues to be a source of controversy. OBJECTIVE: An evaluation is made of the results of treating incisional hernias with a new intra-abdominal low-density composite mesh through both the open and the laparoscopic approach. PATIENTS AND METHODS: A prospective analysis was made on the first 50 patients operated upon for incisional hernia through the open (n=20) or laparoscopic route (n=30), with intra-abdominal repair using the Proceed composite mesh, composed of low-density polypropylene and a hydrophilic antiadherent membrane of oxidized regenerated cellulose. RESULTS: There were no patient deaths. Reintervention proved necessary in one case due to hemoperitoneum caused by a trocar. The mean duration of stay was 3 days, and all patients recovered bowel transit within 24 hours. During follow-up there were no intra abdominal complications associated with the use of the mesh (intestinal occlusion or subocclusion, prolonged ileus, infections, rejection, fistulas, or relapses). CONCLUSIONS: Incision hernia repair using the intra-abdominal low-density composite mesh is safe and well tolerated. Proceed mesh facilitates laparoscopic hernioplasty maneuvering. PMID- 20027096 TI - Totally extraperitoneal laparoscopic hernioplasty: the optimal surgical approach. AB - INTRODUCTION: The total extraperitoneal (TEP) laparoscopic repair of a groin hernia is increasingly being used because of less pain, rapid recovery, and low recurrence rate, but different variants of surgical approaches for TEP are reported by a number of centers and the merits and demerits associated with each approach are not well described. The aim of this study was to compare the clinical outcome of laparoscopic totally extraperitoneal inguinal hernioplasty (TEP) with 4 different variants of surgical approach. METHODS: Between August 2004 and March 2008, 99 patients with unilateral inguinal hernia who underwent TEP without mesh fixation through 4 different variants of surgical approach depended on the anatomical positions of abdominal wall were enrolled in this prospective randomized study. The primary endpoints were operative time, incidence of peritoneal tear, and incidence of the arcuate line impeding the position of the mesh. Secondary endpoints were postoperative analgesic requirements and incidence of seroma. RESULTS: Ninety-nine patients underwent 4 surgical approaches, including the midline approach between the rectus muscle and the posterior rectus sheath (anterior to the posterior rectus sheath, MR) in 25 patients, the midline approach between the peritoneum and the posterior rectus sheath (posterior to the posterior rectus sheath, MP) in 25 patients, the lateral approach between the rectus muscle and the posterior rectus sheath (LR) in 25 patients, and the lateral approach between the peritoneum and the posterior rectus sheath (LP) in 24 patients. The groups were similar in age, weight, body height, and diagnostic categories of hernia. All cases were operated successfully without conversion to open surgery or transabdominal preperitoneal, and followed up for 6 to 43 months with no recurrence. The mean operating time was 55.5 minutes (25 to 130 min) and there was not significant difference between the groups in the operative time. The only type of complication was seroma formation that occurred in 20 patients retrieved without requiring drainage, and there was no significant difference in the incidence of seroma among 4 groups. Only in the MR group and the LR group, the arcuate line in 20 patients impedes the 15 x 10 cm polypropylene mesh positioning. Peritoneal tears were observed without routine closure in 36.7% of patients in the 2 groups of the lateral approach, whereas in only 12% in the 2 groups of the midline approach (P<0.01). Eleven patients required postoperative analgesics, and there was significant difference between the lateral approach groups and the midline approach groups (P<0.05). CONCLUSIONS: TEP is a mature technique and may be safely carried out with fewer complications and lower recurrence rate. Considering the body habitus of each patient and the large dissection surface area resulting in postoperative adhesions and bleeding, according to the principles of "Minimally Invasive Surgery" and the requirement of a maximal cosmetic result, the surgeon should select the midline approach between the peritoneum and the posterior rectus sheath as much as possible. PMID- 20027097 TI - Does single-dose preoperative dexamethasone minimize stress response and improve recovery after laparoscopic cholecystectomy? AB - BACKGROUND: Stress response after laparoscopic cholecystectomy (LC) is less compared with open cholecystectomy, but is still responsible for significant postoperative morbidity. Though preoperative glucocorticoids were found to be effective in reducing the response in open surgical procedures, their role in minimal access surgery is not clear. AIMS AND OBJECTIVES: To evaluate the efficacy of single-dose preoperative dexamethasone in reducing the stress response and postoperative morbidity after LC. MATERIALS AND METHODS: In a prospective randomized, double-blind, placebo-controlled trial, 70 patients undergoing elective LC were randomized to receive either dexamethasone (8 mg intravenously), or placebo. The change in C-reactive protein levels after LC, pain scores at rest, and on exertion and narcotic requirements, the incidence and severity of postoperative nausea and vomiting (PONV), anti-emetic requirement, peak expiratory flow rate in both groups were compared. RESULTS: Dexamethasone was more effective in controlling late PONV (P=0.05). The antiemetic requirement was significantly less in the dexamethasone group (0.56 mg vs. 2.24 mg; P=0.02). Median pain scores were significantly less in the dexamethasone group at 24 hours at rest (P=0.002) and on exertion at 24 and 48 hours (P=0.03 and 0.001). Analgesic requirement was less in the test group (22.9 mg vs. 29.9 mg; P=0.054). The peak expiratory flow rate at 48 hours was higher in the dexamethasone group (315.28 vs. 285.8 l/min; P=0.04). The dexamethasone group showed significantly less elevation of C-reactive protein levels at 24 hours (7.17 microg/mL vs. 17.53 microg/mL; P=0.003) and 48 hours (10.65 microg/mL vs. 23.18 microg/mL; P=0.02) postoperatively. CONCLUSIONS: Preoperative single-dose dexamethasone significantly reduces the pain scores, PONV, and antiemetic requirements while improving the respiratory function in the postoperative period after LC. PMID- 20027098 TI - Trocar site adhesions after laparoscopic surgery in children. AB - PURPOSE: To present our experience of the occurrence of trocar site adhesions in children following laparoscopic procedures. METHODS: Retrospective case notes review of children who were found to have trocar site adhesions at repeat laparoscopy over a 10-year period between 1998 and 2008. RESULTS: All 26 patients who had 28 episodes of repeat laparoscopic procedures were included in the study. There were 11 different procedures. Patient demographics and time interval between the procedures were noted. Trocar site to omentum or bowel adhesions were observed in 3 (10.7%) of the 28 episodes of laparoscopy. In 1 case (3.6% of the total), there were symptoms of small bowel obstruction that required laparoscopic adhesiolysis. The occurrence of adhesions following laparoscopic procedures was found to increase with the number of trocars used. CONCLUSIONS: Our results buttress the claim that the risk of adhesion formation after laparoscopic surgery in children is low. The reduction in the number of trocars used seems to further reduce this risk. PMID- 20027099 TI - The role of laparoscopy in the management of adnexal lesions in children. AB - AIM: Laparoscopy has replaced laparotomy in a variety of surgical abdominal conditions. In pediatric patients the vast majority of adnexal lesions are benign. The aim of this study was to assess the safety and feasibility of laparoscopy in adnexal pathology in children. MATERIALS AND METHODS: The case notes of girls with confirmed adnexal disorders treated in a single center between 1998 and 2008 were reviewed retrospectively. Demographic data, clinical and imaging features, surgical findings and procedures, pathologic features, complications, and outcomes were recorded. RESULTS: Over a 10-year-period 21 patients underwent laparoscopic surgery. Median age at operation was 14 years (range: 2 d to 16 y). Laparoscopy was diagnostic in 1 patient whereas it was therapeutic in 14; 2 patients underwent a laparoscopy-assisted minilaparotomy. The procedure was converted to open in 4 patients due to technical difficulties. The most commonly performed procedures laparoscopically were cyst aspiration (n=6), cystectomy (n=4), unilateral oophorectomy (n=2), and cyst aspiration with deroofing (n=2). No operative or postoperative complications occurred. The overall mean postoperative length of stay was 2.3 days (range: 1 to 6 d); it was 1.8 days (range: 1 to 3 d) for patients who had laparoscopic procedures. Pathology revealed benign lesions in all cases. CONCLUSIONS: In children with benign adnexal pathology, minimally invasive surgery is a feasible and safe approach. Additional benefits of shorter hospital stay, superior cosmetic result, and lesser risk of infertility, make laparoscopy a method of choice for managing pediatric patients. PMID- 20027101 TI - Robotic thyroid surgery: need for initial stricter patient selection criteria. PMID- 20027102 TI - Private sector contributions to pharmaceutical science: thirty-five summary case histories. AB - Expanding government purchases of prescription medicines increase the likelihood of public policies constraining prices and/or the formulary choices available to the beneficiaries of government programs. This can be predicted to reduce private sector incentives for the research and development of new and improved medicines. One response to that argument has been the premise that most of the important scientific advances that yield new and improved medicines do not result from private sector research, but instead are the fruits of research efforts financed or conducted by public agencies, the National Institutes of Health foremost among them. This study addresses that argument by examining the development histories of 32 drugs and drug classes deemed important in the scholarly literature along with three additional specific drugs that have figured prominently in the public discussion of the role of the private sector in drug development. We find that for the discovery and/or development of virtually all of the 32 drug classes, the scientific contributions of the private sector were crucial; and the same is true for the three drugs that have received widespread attention. All or almost all of the drugs discussed would not have been developed-or, at best, would have been delayed significantly-in the absence of private sector scientific discoveries. More generally, both National Institutes of Health-sponsored and private sector pharmaceutical research are crucial for the advancement of pharmaceutical science and the development of new and improved medicines. PMID- 20027103 TI - Psoriasis induced by losartan therapy: a case report and review of the literature. AB - Psoriasis is a papulosquamous disease of multifactorial etiology. A combination of genetic and environmental agents is implicated in its pathogenesis. A variety of triggers, including infection, stress, and medications, have been recognized as precipitants of this disease. Nonsteroidal anti-inflammatory drugs, beta blockers, lithium, synthetic antimalarials, and gold are the most common drugs implicated in precipitating psoriasis. We report a patient with psoriasis induced by initiation of losartan therapy, which resolved with discontinuation of the drug. The Naranjo adverse drug reaction probability scale score indicated that the association between losartan use and psoriasis was probable. PMID- 20027104 TI - Transthoracic echocardiography-guided biopsy of a metastatic endometrial adenocarcinoma in the right atrium: a review of diagnosis and treatment of cardiac masses. AB - We report on a 64-year-old African-American female with a past medical history of an endometrial adenocarcinoma with metastasis to the right atrium. To our knowledge, there have been only four case reports on endometrial carcinoma metastasizing to the right atrium. In our patient, a percutaneous biopsy under fluoroscopic and transthoracic echocardiographic guidance was performed. Histopathologic evaluation of the specimens revealed an adenocarcinoma, consistent with the patient's history of endometrial carcinoma. This report provides a brief review of diagnosis and treatment of cardiac masses. PMID- 20027105 TI - A single supratherapeutic dose of rolofylline does not prolong the QTcF interval in healthy volunteers. AB - Rolofylline is a potent, selective adenosine A1 receptor antagonist that was under development for the treatment of patients with acute decompensated heart failure and renal function impairment. The 30-mg dose of rolofylline administered by intravenous infusion over 4 hours for 3 days represented the anticipated recommended clinical regimen of rolofylline. This was a randomized, double-blind, double-dummy, placebo-controlled, three-period crossover study performed with a single 2-hour intravenous infusion of 60 mg rolofylline, placebo, or oral moxifloxacin in healthy subjects. Plasma samples were collected for determination of rolofylline, M1-trans, and M1-cis pharmacokinetic parameters. The upper limit of the two-sided 90% confidence interval for the placebo-adjusted least squares mean change from baseline in QTcF interval for rolofylline was less than 5 msec at every time point. Moxifloxacin demonstrated an increase in QTcF of greater than 10 msec at 2, 2.5, and 3 hours postdose, thus establishing the sensitivity of the assay to detect modest increases in QTcF interval. Mean Cmax values of 1947.4, 739.2, and 54.8 nM were attained for rolofylline and its metabolites M1 trans and M1-cis, respectively, which were 2.2- to 3.1-fold higher than historic Cmax values seen at the anticipated clinical dose and regimen. Adenosine A1 receptor antagonism from a single supratherapeutic intravenous dose of 60 mg rolofylline over 2 hours was generally well tolerated and did not prolong the QTcF interval relative to placebo. PMID- 20027106 TI - Dexmedetomidine controls the hemodynamic manifestations of tourniquet pain. AB - Dexmedetomidine is an alpha2-adrenergic agonist that is Food and Drug Administration-approved for the provision of short-term (less than 24 hours) sedation of adults during mechanical ventilation. Given its beneficial physiological effects, including sedation and anxiolysis, various applications have been reported in the perioperative period. We report the use of dexmedetomidine to control the hemodynamic consequences of tourniquet pain in three patients having either orthopedic or plastic surgical procedures requiring the use of tourniquets. In all cases, dexmedetomidine controlled the hemodynamic responses to tourniquet pain, including tachycardia and hypertension. The use of dexmedetomidine in this scenario is discussed, and its potential advantages over other agents, including opioids, are reviewed. PMID- 20027107 TI - Successful use of biweekly gemcitabine plus nab-paclitaxel in two male patients with stage iv breast cancer: case reports and review of the literature. AB - Male breast cancer is a rare disease. As a consequence, male breast cancer is often recognized later, and most patients present at an advanced clinical stage. We report the cases of two men with stage IV hormone receptor positive breast cancer who had both received at different times both systemic endocrine therapy with an aromatase inhibitor and gemcitabine as well as nab-paclitaxel-based combination chemotherapy. Although the aromatase inhibitors such as anastrozole, exemestane, and letrozole are very active in postmenopausal women with hormone receptor positive breast cancer, their efficacy in male breast cancer has not been demonstrated in phase II or III trials. Moreover, Gemcitabine and nab paclitaxel every 14 days, with or without bevacizumab, are an active combination in male metastatic breast cancer and should be considered as an option in patients with extensive visceral metastases or hormone refractory disease. PMID- 20027108 TI - The effects of multiple doses of rolofylline on the single-dose pharmacokinetics of midazolam in healthy subjects. AB - Rolofylline is a potent, selective adenosine A1 receptor antagonist that was under development for the treatment of patients with acute decompensated heart failure and renal function impairment. This was a phase I, randomized, open label, 2-period, fixed-sequence study in 19 healthy adult volunteers to examine the effect of multiple intravenous rolofylline doses on the single-dose pharmacokinetics of midazolam, a sensitive CYP3A4 substrate. In period 1, subjects received a single oral dose of midazolam 7.5 mg on day 1. In period 2, subjects received 30 mg, 4-hour infusions of rolofylline (intended clinical dose and duration) once daily for 4 consecutive days; midazolam 7.5 mg was coadministered on day 4. The geometric mean ratios and 90% confidence intervals for AUC0-infinity and Cmax of midazolam in the presence/absence of rolofylline were 1.20 (1.12-1.29) and 1.17 (1.03-1.32), respectively. The apparent terminal half-life (t1/2) for midazolam was similar in the presence/absence of rolofylline (4.31 and 4.27 hours, respectively). The geometric mean ratios (90% confidence intervals) for AUC0-infinity and Cmax of 1'-hydroxymidazolam in the presence/absence of rolofylline were 1.04 (0.96-1.13) and 0.98 (0.84-1.14), respectively. The t1/2 for 1'-hydroxymidazolam was slightly higher in the presence relative to absence of rolofylline (4.24 and 3.17 hours, respectively). Multiple doses of intravenous rolofylline 30 mg for 4 days were generally well tolerated and did not result in clinically important inhibition of CYP3A4 as indicated by little or no change in the pharmacokinetics of midazolam. PMID- 20027109 TI - Comparison of low molecular weight heparin with unfractionated heparin during percutaneous coronary interventions: a meta-analysis. AB - To conduct a meta-analysis of the current evidence to evaluate the safety and efficacy of low molecular weight heparin (LMWH) as compared to unfractionated heparin (UFH). Several studies have demonstrated the therapeutic advantage of LMWH over UFH in the medical management of acute coronary syndromes. However, evidence comparing the 2 in percutaneous coronary interventions (PCI) is inconclusive. Previously published meta-analysis did not include some large-scale trials. We performed a systematic literature search for randomized clinical trials that compared LMWH and UFH in urgent or elective PCI. Studies that evaluated efficacy end points [composite of nonfatal myocardial infarction (MI) and death with or without target vessel revascularization] and bleeding end points were included. Studies were excluded if they involved coadministration of thrombolytics. Data were extracted on an intention-to-treat basis. Heterogeneity of the studies was analyzed by Cochran Q statistics. The Mantel-Haenszel fixed effect model was used to calculate combined relative risks for outcomes where studies were homogenous and the random effect model when the studies were heterogenic. Fourteen studies involving 12,394 patients were included. The efficacy and bleeding risk of LMWH in patients undergoing PCI were comparable with UFH. A subgroup analysis of studies using intravenous or intraarterial administration of LMWH, demonstrated them to be safer than UFH with comparable efficacy. LMWH is at least as efficacious and safe as UFH in patients undergoing PCI. Additionally, evidence suggests that LMWH, when used intravenously, is associated with lower bleeding risks when compared with UFH. PMID- 20027110 TI - Coronary revascularization strategy for ST elevation myocardial infarction with multivessel disease: experience and results at 1-year follow-up. AB - Primary percutaneous coronary intervention (PCI) of culprit lesions (CLs) is the standard of care in patients presenting with ST elevation myocardial infarction (STEMI). However, optimal revascularization strategy for significant nonculprit lesions (non-CLs) in the setting of STEMI remains controversial. The importance of defining of such a strategy lies in the fact that approximately 50% of patients with STEMI have multivessel disease (MVD). The aim of this study was to describe characteristics, therapeutic strategies, and 1-year outcomes in a cohort of patients with STEMI and MVD. We retrospectively analyzed a cohort of 63 patients with STEMI and MVD obtained from a 5-year catheterization database. MVD was defined as >=70% stenosis of >=2 epicardial coronary arteries. This cohort was followed for a period of 1 year for major adverse cardiac events (MACE was defined as acute coronary syndrome, new onset heart failure, or death) and all cause mortality. PCI with stent placement was the major therapeutic procedure (87.5%) performed for CLs. Non-CLs did not undergo interventions in a majority of individuals (47.6%), while the remaining patients underwent PCI (29%) and coronary artery bypass graft surgery (22%) for non-CLs. At 1-year follow-up, prevalence of MACE events and death in the entire cohort were 30% and 15%, respectively. A trend for better outcomes (1-year cumulative MACE events but not mortality) was observed in CL-only intervention cohort compared with non-CL intervention. The PCI and Coronary artery bypass graft surgery cohorts did not show any significant difference in clinical outcomes. In this retrospective cohort of patients with MVD who presented with STEMI, no intervention of noncritical lesions was the prevalent approach, reflecting guideline recommendations. CL-only intervention strategy showed a better clinical outcome than non-CL intervention. Intervention of noncritical lesions therefore did not seem to improve MACEs or all-cause mortality at 1-year of follow-up and might in fact have had a detrimental effect on outcomes. PMID- 20027112 TI - Clinical and genetic aspects of neurofibromatosis 1. AB - Neurofibromatosis 1 is an autosomal dominant disorder characterized by multiple cafe-au-lait spots, axillary and inguinal freckling, multiple cutaneous neurofibromas, and iris Lisch nodules. Learning disabilities are present in at least 50% of individuals with neurofibromatosis 1. Less common but potentially more serious manifestations include plexiform neurofibromas, optic nerve and other central nervous system gliomas, malignant peripheral nerve sheath tumors, scoliosis, tibial dysplasia, and vasculopathy. The diagnosis of neurofibromatosis 1 is usually based on clinical findings. Neurofibromatosis 1, one of the most common Mendelian disorders, is caused by heterozygous mutations of the NF1 gene. Almost one half of all affected individuals have de novo mutations. Molecular genetic testing is available clinically but is infrequently needed for diagnosis. Disease management includes referral to specialists for treatment of complications involving the eye, central or peripheral nervous system, cardiovascular system, spine, or long bones. Surgery to remove both benign and malignant tumors or to correct skeletal manifestations is sometimes warranted. Annual physical examination by a physician familiar with the disorder is recommended. Other recommendations include ophthalmologic examinations annually in children and less frequently in adults, regular developmental assessment in children, regular blood pressure monitoring, and magnetic resonance imaging for follow-up of clinically suspected intracranial and other internal tumors. PMID- 20027113 TI - Maternal systemic primary carnitine deficiency uncovered by newborn screening: clinical, biochemical, and molecular aspects. AB - BACKGROUND: Systemic primary carnitine deficiency is an autosomal recessive disorder of the carnitine cycle caused by mutations in the SLC22A5 gene that encodes the carnitine transporter, organic cation transporter. Systemic primary carnitine deficiency typically presents in childhood with either metabolic decompensation or cardiomyopathy. We report five families in which low free carnitine levels in the infants' newborn screening have led to the diagnosis of maternal systemic primary carnitine deficiency. METHODS: Blood samples from the infants and /or their family members were used to extract the DNA. The entire coding regions of the SLC22A5 gene were sequenced. The clinical data were obtained from the referring metabolic specialists. RESULT: Sequencing the SLC22A5 gene allowed molecular confirmation with identification of three novel mutations: c.1195C>T (p.R399W), c.1324_1325GC>AT (p.A442I), and c.43G>T (p.G15W). All infants were asymptomatic at the time of diagnosis, and one was found to have systemic primary carnitine deficiency. Three mothers are asymptomatic, one had decreased stamina during pregnancy, and one has mild fatigability and developed preeclampsia. DISCUSSION: These findings provide further evidence that systemic primary carnitine deficiency presents with a broad clinical spectrum from a metabolic decompensation in infancy to an asymptomatic adult. The maternal systemic primary carnitine deficiency was uncovered by the newborn screening results supporting the previous notion that newborn screening can identify some of the maternal inborn errors of metabolism. It also emphasizes the importance of maternal evaluation after identification of a low free carnitine level in the newborn screening. PMID- 20027111 TI - Review of studies on metabolic genes and cancer in populations of African descent. AB - Genetic polymorphisms described for a number of enzymes involved in the metabolism of tobacco carcinogens and alcohol have been linked to increase cancer risk. Racial disparities in cancer between whites and populations of African descent are well documented. In addition to differences in access to health care, both environment and genetic factors and their interaction may contribute to the increased cancer risk in minority populations. We reviewed the literature to identify case-control studies that included subjects of African descent. Meta analyses investigating the association of genetic polymorphisms in tobacco metabolic genes and cancer were performed. Although several genes and cancers have been studied, only one or two studies per gene for each cancer site have been published, with the exception of breast (CYP1A1 and CYP1B1), lung (GSTM1, CYP1A1, and NQO1), and prostate (CYP3A4 A293G and CYP17). Marginal statistically significant associations were observed for CYP3A4 A293G and CYP17 5'UTR polymorphisms and prostate cancer. Our findings support the need for additional genetic association studies of breast, prostate, and lung cancers that include a larger number of minority participants. Because incidence and mortality rates for these cancers rank highest among populations of African descent, concentrated research in these areas are warranted. PMID- 20027114 TI - Hereditary breast and ovarian cancer: referral source for genetic assessment and communication regarding assessment with nongenetic clinicians in the community setting. AB - PURPOSE: To examine referral source to cancer genetic services; communication of results of genetic evaluation to clinicians; role of clinicians in postcounseling management; and use of alternative information sources after cancer genetic risk assessment/counseling in the community setting. METHODS: Retrospective telephone survey. SETTING: A community/private hospital-based cancer genetic counseling service. PATIENTS: Women, at least 21 years of age, who had undergone cancer genetic counseling with (1) at least a 10% predicted likelihood of carrying a BRCA1/2 mutation or (2) a documented BRCA1/2 mutation. INTERVENTION: A 121-item telephone survey. MAIN OUTCOME MEASURE: (1) initial referral source to cancer genetic services; (2) women's communication of results of cancer genetic assessment to primary and (nongenetic) specialist clinician(s); (3) education and support role played by subjects' physician(s); and (4) use of other hereditary breast and ovarian cancer (HBOC) information resources. RESULTS: Of 225 women eligible for study, 69 (31%) completed the survey. Sixty-two percent were referred by their medical oncologist; 13% by their primary care physician, and fewer by their surgeon (6%) or gynecologist (4%). Results of the cancer genetic assessment were not shared with 19% of primary care clinicians, 26% of primary gynecologists, 12% of oncologists, and 36% of surgeons. Twenty-six percent of participants noted that their primary care clinician had not been involved in their HBOC-related, cancer prevention decisions, 16% had not included their gynecologist, 2% had not involved their oncologist, and 20% replied that their surgeon had not been involved in these decisions. Overall, clinicians were perceived as supportive when it came to a participants' information and decision support needs. One exception was that 21% of respondents reported the use by clinicians of medical terms, without definition. Over two-thirds had sought alternative "self-help" HBOC-related materials, most Internet based. CONCLUSIONS: These results have implications for interdisciplinary communication and decision support for those with or at risk for HBOC, cared for in the community setting. PMID- 20027115 TI - A validated disease severity scoring system for adults with type 1 Gaucher disease. AB - PURPOSE: A validated disease severity scoring system (DS3) for Gaucher disease type 1 (GD1) is needed to standardize patient monitoring and to define patient cohorts in clinical studies. METHODS: DS3 domains were established by an expert physician group using the nominal group technique of consensus formation. Items were selected by 36 GD1 physicians. The expert group determined appropriate measurement techniques for each item. Measurements were weighted considering contributions to GD1 morbidity and mortality. Consensus Clinical Global Impression Severity scores for sample cases were compared with average DS3 scores. A minimal clinically important difference in GD1 DS3 score was calculated. RESULTS: The GD1 DS3 includes bone (42% of score), hematologic (32%), and visceral domains (26%); individual items use routine assessments, including medical history, blood chemistry, organ volume measurements, and bone evaluations (magnetic resonance imaging and dual x-ray absorptiometry). The maximum score is 19. Interrater reliability was 0.97 (Cohen's kappa). DS3 scores were highly correlated with Clinical Global Impression Severity scores (r2 = 0.89). The minimal clinically important difference was -3.2 improvement and +3.9 deterioration. CONCLUSION: This DS3 accurately quantifies GD1 status and intrapatient change over time. Testing of reliability and validity will continue to allow eventual implementation of the DS3 in clinical studies and routine practice. PMID- 20027116 TI - Dose reduction for semi-automated volumetry of hepatic metastasis in MDCT studies. AB - OBJECTIVES: To investigate the performance of semi-automated measurements (RECIST, volume) of hepatic metastases in multidetector-row computed tomography (MDCT) under normal-dose- and simulated low-dose-protocols. MATERIALS AND METHODS: Thirty-five patients (67 +/- 13 years) with a total of 79 hepatic metastases underwent 16-MDCT (120 kv, 160 mAseff, pitch 1, 3 mm slice thickness, 2 mm reconstruction increment, B30f standard soft tissue kernel) for either initial staging or therapy monitoring. Corresponding raw data from these standard dose scans were simulated at lower radiation doses of 80/60/40 mAseff (Somatom Noise Vers.6.1 beta, Siemens Healthcare, Forchheim, Germany). A semi-automated software tool (SyngoCT Oncology, Siemens Healthcare, Forchheim, Germany) was applied to each dose setting to evaluate size parameters (RECIST, volume). These measurements were compared by applying repeated-measures analysis of variance and displayed graphically. RESULTS: For RECIST measurements no statistically significant differences were found between standard dose (Mean RECIST diameter: 20.46 +/- 8.37 mm) and different simulated low radiation doses (80 mAseff: 20.95 +/- 8.20 mm/60 mAseff: 20.50 +/- 8.35 mm/40 mAseff: 19.95 +/- 8.16 mm): P = 0.0774.Statistically significant differences of volume quantification (P < 0.05) could be found between standard-(3.60 +/- 4.63 mL) and simulated lowest dose of 40 mAseff (3.17 +/- 4.08 mL), whereas there was no difference (P > 0.05) between 160 mAseff- and either 80 mAseff-(3.46 +/- 4.31 mL) or 60 mAseff-protocols (3.44 +/- 4.35 mL). CONCLUSIONS: Software-assisted assessment of RECIST criteria and volume demonstrated valid performances under different dose-settings in MDCT; therefore, substantial radiation dose reduction could be possible with the use of semi-automated measurements in follow-up studies. PMID- 20027117 TI - Field strength and diffusion encoding technique affect the apparent diffusion coefficient measurements in diffusion-weighted imaging of the abdomen. AB - OBJECTIVES: The purpose of this study is to determine what effects a variety of diffusion encoding techniques at 1.5 T and 3 T have on measured abdominal apparent diffusion coefficient (ADC) values obtained in a healthy population. MATERIALS AND METHODS: Sixteen healthy male volunteers were enrolled in this prospective Institutional Review Board-approved study following written informed consent. Imaging was performed on a 1.5 T and a 3 T magnetic resonance system (Siemens, Erlangen) with several abdominal axial diffusion weighted imaging (DWI) acquisitions: an orthogonal diffusion encoding with b-values of 0/400 seconds/mm, and a series of four 3-scan trace weighted acquisitions with b-values of 0/50, 0/400, 0/800, 0/50/400/800 seconds/mm, respectively. The mean ADC values were calculated for 3 regions of interest (ROI) in 5 locations (right hepatic lobe, spleen, pancreatic head, body, and tail). The ADC data were analyzed using a repeated-measures analysis of variance. RESULTS: There was a significant difference between measured ADC values at 1.5 T and 3 T for liver (P < 0.001), but not for pancreas (P = 0.427) or spleen (P = 0.167). There was no significant difference (P > 0.999) in the measured ADC values between the orthogonal encodings and the 3-scan trace weighted encoding with the same b-value. There were significant differences (P < 0.001) between all 4 weighting schemes for the 3-scan trace with the measured ADC decreasing with increasing b-value. CONCLUSION: Measured abdominal ADC values depend on the exact selection of b value used for encoding for liver, pancreas, and spleen. In addition, the measured ADC values depend on the field strength of the scanner for liver. PMID- 20027118 TI - BR55: a lipopeptide-based VEGFR2-targeted ultrasound contrast agent for molecular imaging of angiogenesis. AB - OBJECTIVES: BR55, an ultrasound contrast agent functionalized with a heterodimer peptide targeting the vascular endothelial growth factor receptor 2 (VEGFR2), was evaluated in vitro and in vivo, demonstrating its potential for specific tumor detection. MATERIALS AND METHODS: The targeted contrast agent was prepared by incorporation of a biospecific lipopeptide into the microbubble membrane. Experiments were performed in vitro to demonstrate the binding capacities of BR55 microbubbles on immobilized receptor proteins and on various endothelial or transfected cells expressing VEGFR2. The performance of BR55 microbubbles was compared with that of streptavidin-conjugated microbubbles targeted to the same receptor by coupling them to a biotinylated antibody. The specificity of BR55 binding to human and mouse endothelial cells was determined in competition experiments with the free lipopeptide, vascular endothelial growth factor (VEGF), or a VEGFR2-specific antibody. Molecular ultrasound imaging of VEGFR2 was performed in an orthotopic breast tumor model in rats using a nondestructive, contrast-specific imaging mode. RESULTS: BR55 was shown to bind specifically to the immobilized recombinant VEGFR2 under flow (dynamic conditions). BR55 accumulation on the target over time was similar to that of microbubbles bearing a specific antibody. BR55 avidly bound to cells expressing VEGFR2, and the pattern of microbubble distribution was correlated with the pattern of receptor expression determined by immunocytochemistry. The binding of targeted microbubbles on cells was competed off by an excess of free lipopeptide, the natural ligand (VEGF) and by a VEGFR2-specific antibody (P < 0.001). Although selected for the human receptor, the VEGFR2-binding lipopeptide was also shown to recognize the rodent receptor. Tumor perfusion was assessed during the vascular phase of BR55, and then the malignant lesion was highlighted by specific accumulation of the targeted microbubbles on tumoral endothelium. The presence of VEGFR2 was confirmed by immunofluorescence staining of tumor cryosections. CONCLUSIONS: VEGFR2-targeted ultrasound contrast agents such as BR55 will likely prove useful in human for the early detection of tumors as well as for the assessment of response to specific treatments. PMID- 20027119 TI - Assessment of left atrial volumes and function in orthotopic heart transplant recipients by dual-source CT: comparison with MRI. AB - INTRODUCTION: To compare left atrial performance with dual-source CT (DSCT) with respect to magnetic resonance imaging (MRI) in orthotopic heart transplant recipients. METHODS: Twenty-nine consecutive heart transplant recipients (27 male; mean age 64.1 +/- 13 years; mean time from transplantation 122.8 +/- 69.7 months) referred for exclusion of cardiac allograft vasculopathy underwent cardiac DSCT and MRI. Standard biatrial technique was employed in 13 subjects whereas 16 were transplanted after the bicaval technique. Axial 5-mm slice thickness DSCT datasets reconstructed in 5% steps of the cardiac cycle and axial 5-mm SSFP-MRI images were analyzed. Two blinded readers manually traced left atrial contours in random order to estimate end-diastolic volume (EDV), end systolic volume (ESV), and ejection fraction (EF). Parameters were compared with a paired sample Student t test. Concordance correlation coefficient (CCC) was calculated to determine measurement agreement between techniques and observers. RESULTS: Left atrial volumes were significantly higher with cardiac DSCT (EDV: 170.9 +/- 78.1 mL; ESV: 139.5 +/- 76.6 mL) than with MRI (EDV: 158.2 +/- 72.5 mL; ESV: 124.2 +/- 68.2 mL), whereas left atrial EF was lower with DSCT (EF: 20.8% +/ 7.5% vs. 23.6% +/- 7.7%) (P < 0.05). Measurement agreement between DSCT and MRI was excellent for all parameters (CCC > or =0.82). Individuals operated with the biatrial anastomosis technique presented significantly higher left atrial volumes and lower EF compared with subjects with bicaval anastomosis. Interobserver agreement was excellent for all parameters (CCC > or =0.80). CONCLUSION: Even if DSCT slightly overestimates left atrial volumes with respect to MRI, results remain clinically valid. Bicaval surgical technique offers improved left atrial performance compared with standard biatrial anastomosis. DSCT may be used as a reliable tool to estimate left atrial parameters in orthotopic heart transplant recipients. PMID- 20027120 TI - Semiautomatic lymph node segmentation in multislice computed tomography: impact of slice thickness on segmentation quality, measurement precision, and interobserver variability. AB - OBJECTIVES: To determine the impact of slice thickness on semiautomatic lymph node analysis. MATERIALS AND METHODS: Thoracic multislice computed tomography (MSCT) of 46 patients with NSCLC were reconstructed at 1.0-, 3.0-, and 5.0-mm slice thickness. Two radiologists independently determined long and short axis diameter and volume of 299 thoracic lymph nodes by semiautomatic segmentation software. Necessity of manual correction (= relative difference between uncorrected and corrected segmented lymph node volume) and relative interobserver differences were determined. The precision of segmentation was expressed by relative measurement deviations (RMD) from the reference standard (mean of 1.0 mm datasets). Statistical analysis encompassed t test and Bland-Altman plots. RESULTS: Necessity of manual correction was significantly higher for 5.0 mm than for 3.0 (P = 0.042) or 1.0 mm (P = 0.0012). The RMD for long and short axis diameter were found to be independent of slice thickness, whereas the RMD for lymph node volume significantly (P = 0.021) increased from 4.0% at 1.0 mm (95% CI: 1.0%-3.5%) to 35% at 5.0 mm (95% CI: 10.5%-60.5%). The relative interobserver differences was consistently low for metric and volumetric parameters (eg, volume 2.3%, 95% CI: -7.4%-10.8% at 5.0 mm) with no difference in any of the slice thicknesses (P > 0.064). CONCLUSIONS: Significant deviations in lymph node volume together with excessive manual corrections suggest reconstruction of the data for semiautomatic lymph node assessment at a slice thickness of 1.0 mm but not exceeding 3.0 mm. PMID- 20027121 TI - Saving dose in triple-rule-out computed tomography examination using a high-pitch dual spiral technique. AB - OBJECTIVE: High radiation doses remain a drawback of current triple-rule-out computed tomography (CT) protocols. With dual source CT, a new high-pitch dual spiral technique offers the possibility to acquire an Electrocardiography (ECG) gated-synchronized dataset of the whole chest in less than 1 second. The aim of this study was to compare the dose of such a protocol to a standard, nongated chest scan and to a conventional, retrospectively ECG-gated triple-rule-out protocol. Also, the efficacy and dose of this dual spiral protocol was to be compared in patients examined with this high-pitch technique and matched controls scanned with the conventional technique. MATERIALS AND METHODS: An anthropomorphic Alderson Rando phantom was equipped with thermoluminescent detectors and scanned with the high-pitch protocol (Siemens Somatom Definition Flash; 2 x 120 kVp, 426 mAseff, 128 x 0.6 mm collimation, pitch 3.2), the nongated chest scan (same scanner; 120 kVp, 160 mAseff, 128 x 0.6 mm, pitch 1.2; equivalent Computed Tomography Dose Index (CTDI) of 7.12 mGy), and the conventional gating technique (Siemens Somatom Definition; 120 kVp, 560 mAseff with ECG pulsing interval at 30%-70% of the R-R cycle, 64 x 0.6 mm, pitch 0.3). Noise was measured in air, central and peripheral soft tissue of the phantom. Conversion factors were determined based on the measured dose and the dose-length products of the scanner. The protocol was then applied with ethics committee approval in 31 patients suffering from acute chest pain. The 120 mL of contrast material (Ultravist 370, Bayer Schering Pharma) was applied at 5 mL/s. Dose was calculated based on the dose-length products and the conversion factor. Image quality was assessed by 2 readers for aorta, pulmonary arteries, and coronary arteries. The results were compared with matched controls scanned with the conventional ECG gating technique and non-ECG gated thorax scans. RESULTS: The dose determined with thermoluminescent dosimeters measurements amounted to 2.65, 2.68, and 19.27 mSv, respectively, for the dual spiral technique, the standard chest scan, and the conventional retrospective technique. There was no significant difference in image noise. Respective conversion factors were 0.0186, 0.0188, and 0.0180 mSv/mGy x cm. In the patient examinations, dose was 4.08 +/- 0.81 mSv with the high-pitch protocol compared with 20.4 +/- 5.3 mSv in the matched controls with the conventional technique, and 4.40 +/- 0.83 mSv for the non-ECG gated thorax scan. Scan times were 0.7 +/- 0.1 seconds for the high-pitch scan and 15 +/- 3 seconds for the conventional chest pain scan. Aorta and pulmonary arteries were depicted in diagnostic quality in both groups. About 84.7% of coronary artery segments were rated as diagnostic in the high-pitch exams (95.4% below 65 bpm and only 72.8% in higher heart rates), whereas 92.9% were diagnostic with the conventional approach. CONCLUSION: The high-pitch dual spiral technique requires only about one-fifth of the dose of conventional ECG gated triple-rule-out protocols, thus matching that of a standard nongated chest scan. With less than 1 second, the scan time is very short. This protocol can be recommended for patients with unclear chest pain with rhythmic heart rates below 65 bpm. PMID- 20027122 TI - Common genetic variations of the renin-angiotensin-aldosterone system and response to acute angiotensin I-converting enzyme inhibition in essential hypertension. AB - OBJECTIVE: In order to get insight into possible genetic determinants of antihypertensive drug action, we analysed the relations between polymorphisms of the genes of the renin-angiotensin-aldosterone system and acute effects of ACE inhibition on blood pressure as well as circulating renin and aldosterone levels in hypertensive patients. METHODS: A total of 315 hypertensive patients referred for problems in drug treatment were given a single 50 mg dose of captopril. Plasma renin and aldosterone were measured before and 60 min after the drug administration. Four DNA variants, including angiotensin type I receptor (AGTR1) 1166 A/C, angiotensin-converting enzyme (ACE) I/D, angiotensinogen (AGT) M235T and AGT -217 G/A, were genotyped in the patients and normotensive men (n = 175). A replication study on the relation between AGTR1 1166 A/C and plasma renin and aldosterone levels was carried out in the 244 hypertensive men of the pharmacogenetic GENRES Study. RESULTS: Referred hypertensive patients with the AGTR1 CC genotype had higher aldosterone at baseline (P = 0.02) and after 60 min of captopril administration (P = 0.01) compared with the AA genotype. Replicate analysis in the GENRES patients showed a similar trend. When the two studies were combined (315 and 244 patients, respectively), plasma aldosterone level (P = 0.007) as well as aldosterone/renin ratio (P = 0.04) were significantly higher in the CC genotype (n = 13) than in the AA genotype (n = 370). Transfection studies in cultured HEK293 cells indicated that the 1166C allele was associated with higher mRNA levels than the 1166A allele. CONCLUSION: The AGTR1 1166C allele when present in homozygous form may be associated with a form of essential hypertension characterized by high plasma aldosterone and low plasma renin levels, possibly due to increased AGTR1 mRNA levels and augmented angiotensin II action. PMID- 20027124 TI - Adjuvant cisplatin and vinorelbine for completely resected non-small cell lung cancer: subgroup analysis of the Lung Adjuvant Cisplatin Evaluation. AB - BACKGROUND: To evaluate the impact of adjuvant cisplatin-vinorelbine in completely resected non-small cell lung cancer and identify patients likely to benefit from this regimen in the Lung Adjuvant Cisplatin Evaluation (LACE) database. The overall LACE meta-analysis showed survival benefit with cisplatin based adjuvant chemotherapy (5-year survival benefit of 5.4%, hazard ratio [HR] 0.89, p = 0.004). Subgroup analysis for the cisplatin-vinorelbine regimen was prespecified in the LACE statistical analysis plan. Patients randomized to cisplatin-vinorelbine or observation were the largest subgroup (41%) and the most homogeneous in terms of drug doses and eligibility. PATIENTS AND METHODS: The LACE-vinorelbine cohort included trials evaluating cisplatin-vinorelbine versus observation. Overall survival was the primary end point. Other studies randomizing patients to other chemotherapy or observation (LACE-other) were also evaluated. RESULTS: The LACE-vinorelbine cohort included 1888 patients from four studies (Adjuvant Navelbine International Trialist Association, Big Lung Trial, International Adjuvant Lung Cancer Trial, and National Cancer Institute of Canada Clinical Trials Group JBR.10). Baseline characteristics were similar to the LACE other but had fewer patients with stage IA (2% versus 11%). Survival improvement at 5 years was 8.9% with cisplatin-vinorelbine versus observation (HR 0.80, 95% confidence interval: 0.70-0.91, p <0.001). Stage was a significant predictor for survival (test for trend, p = 0.02; benefit at 5 years: 14.7% [stage III], 11.6% [stage II], and 1.8% [stage I]). Similar benefits were seen for disease-free survival (HR 0.75 [0.67-0.85, p <0.001], stage III [HR 0.62, 0.50-0.76], stage II [HR 0.69, 0.57-0.83], and stage I [HR 0.95, 0.767-1.19]). The overall result was statistically superior to LACE-other (LACE other HR 0.95, 0.86-1.05, interaction p = 0.04). CONCLUSION: In subgroup analyses, adjuvant cisplatin-vinorelbine provides a superior survival benefit and can be recommended in completely resected stages II and III non-small cell lung cancer. PMID- 20027125 TI - Sequence types and antimicrobial susceptibility of invasive streptococcus pneumoniae isolates from a region with high antibiotic selective pressure and suboptimal vaccine coverage. AB - Multilocus sequence typing was carried out on 95 invasive pneumococcal isolates belonging to the most common 7 serotypes currently circulating in Taiwan. The study confirmed continued prevalence in Taiwan of a few global clones and sequence types (STs) since the mid-1990s and identified the recent emergence of ST320 (19A) and ST902 (6A). Antimicrobial nonsusceptibility was common in the predominant STs of serotypes 14, 19A, 19F, and 23F. PMID- 20027123 TI - Centrally administered lipopolysaccharide elicits sympathetic excitation via NAD(P)H oxidase-dependent mitogen-activated protein kinase signaling. AB - OBJECTIVE: The mechanisms by which inflammation activates sympathetic drive in heart failure and hypertension remain ill-defined. In this study, an intracerebroventricular injection of lipopolysaccharide (LPS) was used to induce the expression of cytokines and other inflammatory mediators in the brain, in the absence of other excitatory mediators, and the downstream signaling pathways leading to sympathetic activation were examined using intracerebroventricular injections of blocking or inhibiting agents. METHODS AND RESULTS: In anesthetized rats, intracerebroventricular injection of LPS (5 microg) increased (P < 0.05) renal sympathetic nerve activity, blood pressure and heart rate. LPS increased (P < 0.05) hypothalamic mRNA for NAD(P)H oxidase subunits p47 and gp91, NAD(P)H oxidase-dependent superoxide generation, hypothalamic mRNA for tumor necrosis factor-alpha, cyclooxygenase-2 and cerebrospinal fluid levels of tumor necrosis factor-alpha and prostaglandin E2. In the paraventricular nucleus of hypothalamus, dihydroethidium staining for superoxide expression and c-Fos activity (indicating neuronal excitation) increased. The superoxide scavenger tempol significantly (P < 0.05) diminished the expression of inflammatory mediators, as well as superoxide expression and neuronal excitation in paraventricular nucleus. SB203580 (p38 mitogen-activated protein kinase inhibitor) also reduced the expression of inflammatory mediators in hypothalamus and cerebrospinal fluid. Tempol, apocynin [NAD(P)H oxidase inhibitor], SB203580 and NS398 (cyclooxygenase-2 inhibitor) all reduced cerebrospinal fluid prostaglandin E2 and the sympathoexcitatory response to LPS. LPS also increased angiotensin II type 1 receptor mRNA, a response blocked by apocynin and tempol but not by SB203580. CONCLUSION: These findings suggest that central inflammation in pathophysiological conditions activates the sympathetic nervous system via NAD(P)H oxidase and p38 mitogen-activated protein kinase-dependent synthesis of prostaglandin E2. PMID- 20027126 TI - Role of psychosocial factors in the etiology of temporomandibular disorders: relevance of a biaxial diagnosis. AB - AIM: The prevalence of temporomandibular disorders (TMD) is higher among women than men, indicating a multifactorial role for gender-related differences in the etiology of TMD: physiological hormonal differences, inflammatory response to stress, and sociocultural differences in response to pain. The aim of this study was to draw a biobehavioral picture of the TMD patient based on Research Diagnostic Criteria for TMD (RDC/TM) Axis II diagnosis and analysis of gender related differences. METHODS: Between January 2006 and January 2008, 362 subjects were consecutively enrolled from patients who presented at the Clinic for Temporomandibular Disorders, School of Dental Medicine, University of Pavia, because of orofacial pain, limitation or joint sounds on mandibular movement. Of the 362 subjects evaluated, 308 met the inclusion criteria. RESULTS: The average age of the study population was 41 years; the female: male ratio was 4:1. When stratified according to chronic pain intensity grade and gender, 26% of the women had grade I, 36.4% grade II, 17% grade III, and 9.7% grade IV; 34.4% of the men had grade I, 32.8% grade II, 6.5% grade III, and 3.3% grade IV. Depression was moderate in 35 women and in 6 men and severe in 138 women and in 24 men; somatization was moderate in 59 women and in 20 men and severe in 143 women and in 19 men. CONCLUSIONS: Gender-related differences may be considered risk factors for TMD; psychological characteristics, including somatization, depression, and anxiety related to gender, appear to have a significant impact on the prevalence of TMD. PMID- 20027127 TI - Deglutition and temporomandibular disorders in children. AB - AIM: This study aimed to evaluate the characteristics of deglutition in children having or not temporomandibular disorders (TMD) or signs and/or symptoms of TMD. METHODS: The sample comprised 152 children aged from 8 to 12 years (78 males and 74 females, mean age 10.05+/-1.39 years). The clinical signs were evaluated using the axis I of the Research Diagnostic Criteria for TMD (RDC/TMD) and the symptoms, using a questionnaire. Patients were divided into the following groups: Group TMD (N.=40), signs and symptoms of TMD (Group S and S, N.=68), signs or symptoms of TMD (Group S or S, N.=33) and without signs and symptoms (Group N, N.=11). Characteristics of orofacial structures such as occlusion, tongue, lingual frenulum, lips and mentalis muscle were evaluated. Myofunctional evaluation during deglutition with solid (bread) and liquid (water) was also performed. RESULTS: A high prevalence of abnormal deglutition was found, with similar proportion in groups. Alterations in lips, mentalis muscle and tongue in swallowing was significantly smaller in Group N than in the other groups. The proportions of children with lower lip interposition and lateral tongue thrust, when swallowing liquid, were significantly higher than swallowing solids. There was a smaller proportion of children in Group N with lower lip interposition when swallowing liquids. CONCLUSIONS: TMD or presence of signs and/or symptoms of TMD was not associated with an abnormal deglutition. Nevertheless, orofacial myofunctional alterations could be considered influencing factors on TMD, due to the high prevalence of abnormal deglutition pattern. In addition, the abnormal deglutition could be attributed to the malocclusion, mixed dentition phase and orofacial myofunctional characteristics. PMID- 20027128 TI - Assessment of the polymerization contraction stress of composites photoactivated by halogen light and light-emitting diode. AB - AIM: The aim of this study was to compare the polymerization contraction stress of composites photoactivated by two light sources: quartz tungsten halogen light (QTH) and light emitting diode (LED). METHODS: Glass rods were fabricated (5.0 mm X 5.0 cm) and one of the surfaces was air abraded with aluminum oxide. An adhesive was applied to this surface and photoactivated by LED. The glass rods were assembled to a Universal Testing machine (Instron - 5565) and the composite were applied to the lower rod using a manual instrument. The upper rod was placed closer, at 2 mm, and an extensometer was attached to the rods. The twenty composites were polymerized by either by QTH (N.=10) or LED (N.=10). Polymerization was carried out using two apparatuses positioned in opposite sides, which were simultaneously activated for 30 seconds. Contraction stress was analyzed twice: shortly after polymerization (t30s) and 30 minutes later (t30min). RESULTS: The contraction stress for all composites was higher at t30min than at t30s, regardless of the activation source. Z100 showed lower contraction stress values (P<0.05) compared to the other composites. Regarding to Charisma and TPH, the photoactivation source had no influence on contraction stress, except for Z100 at t30min. CONCLUSIONS: It was concluded that composite composition is the factor that most influence the polymerization contraction stress. PMID- 20027129 TI - Cephalometric evaluation of white Brazilian adult skeleton. AB - AIM: The aim of the present study was to compare cephalometric norms of white Brazilian adults with the values described by Tweed, who measured white North American adults. Sexual dimorphism was also evaluated, and male and female facial features were compared. METHODS: Samples were obtained from 40 lateral cephalograms and divided into groups of 20, according to patient's gender. The radiographs were analyzed using the computer-assisted Tweed cephalometric analysis and the 2.0 Radiocef software. Results were analyzed statistically using the Student's t test for genders and ethnicities. The level of significance was set at P< or =0.05. RESULTS: The comparison with Tweed norms revealed that Brazilians have a convex profile and more marked labial tipping of mandibular incisors than North Americans. Sexual dimorphism was found only in linear measurements, which suggests that facial morphology is similar for Brazilian males and females. CONCLUSIONS: Results revealed significant statistical differences in linear and angular cephalometric dimensions between different ethnic groups, which supports the idea that ethnicity should be included in cephalometric analyses. PMID- 20027130 TI - Microleakage of occlusoproximal adhesive restorations, effect of dentin moisture after acid etching. AB - AIM: The aim of this study was to evaluate the effect of dentin moisture after acid-etching on marginal sealing in occluso-proximal composite restorations. METHODS: Ninety occluso-proximal preparations were made in posterior teeth with gingival margin in dentin. Prepared teeth were divided in 3 groups (N=30) according to the type of solvent of adhesive (ethanol/water=SB, Single Bond 3M ESPE, acetone=PB, Prime & Bond NT Dentsply, and ethanol=OP, OptiBond Solo Plus, Kerr). And 3 subgroups (N=10) according to the types of treatment of the cavity after rinsing the acid etchant ( 1) cavity air-dried for 10 s at a distance of 1 cm; 2) cavity blot-dried with slightly moist cotton pellets; 3) cavity with visible excess of water). Hybrid composite was applied in four increments. After 24 hours, teeth were thermocycled (500 cycles, 30 s, 5 to 55 masculineC) and immersed in 0.5% basic fucsin for 4 hours. The teeth were cleaned, dried and sectioned. The microleakage extent was analyzed with a 0-5 scale in dentin margin. Enamel margins were verified for presence or absence of microleakage. Data were submitted to Kruskal Wallis test (P<0.05). RESULTS: The median of microleakage scores in dentin margins were: SB_a=1, SB_b=0, SB_c=1, PB_a=1, PB_b=1, PB_c=2, OP_a=0, OP_b=1, OP_c=1. Considering bonding systems, PB provided more microleakage than SB and OP (P=0.0316). In groups where cavity had visible excess of water there was higher microleakage degree than the dry cavity subgroups (P=0.048). In enamel margins, median was 0 for all groups. CONCLUSIONS: Better marginal sealing is obtained in dentin margins of occluso-proximal cavities when they are air-dried or blot-dried with cotton pellets after acid etching. PMID- 20027131 TI - The natural head position. Different techniques of head positioning in the study of craniocervical posture. AB - The aim of this paper was to describe the most important definitions of head posture and the most important methodology to get an natural head posture. After a careful analysis of the literature with no limitations of language or time period, 31 papers were selected. Relevant information was also derived from reference lists of the publications retrieved. The key words used in the search were head posture, cranio-cervical posture, cephalometric analysis, natural head position, lateral cephalometric radiographs, cephalostat, self balance position, mirror position. The definitions of natural head position are various. An accurate head posture registration is time consuming and not particularly feasible on a clinical field. Nevertheless it is possible to apply a craniostat to the patient before the execution of the radiograph without modifying the NHP. The execution of radiographs of the craniocervical zone is connected to the correct position of the head. The NHP is a reproducible positions, and can be useful for making comparisons at any time in the case of the same patient or in comparing different patients cephalometrically. PMID- 20027132 TI - Oral hobnail hemangioma: case report. AB - Hobnail hemangioma (HH) is a rare benign vascular neoplasm reported as a distinctive small benign, solitary vascular neoplasm of the superficial and mid dermis occurring on the face, trunk, or extremities of young or middle-aged adults. The oral manifestations are quite uncommon, with only three cases reported in the English language literature. The following case refers to a 38 year-old woman with a small nodule on the hard palatal mucosa. Histological findings showed a biphasic growth pattern of irregularly dilated vascular structures in the superficial mucosa, lined by epithelioid endothelial cells with a hobnail appearance, with neoplastic vessels observed in deeper parts of the lesion. In the oral cavity, the differential diagnosis includes hemangioma, melanoma, and Kaposi's sarcoma, whose clinical and histological features may be confused with those of HH. PMID- 20027133 TI - Fibrolipoma associated with a mucus retention cyst in the palate: a case report. AB - Fibrolipomas are benign mesenchymal neoplasms of the fatty tissue rarely encountered in the oral cavity. They account for around 1% to 5% of all neoplasms affecting the mouth and occur as raised, slow-growing, painless lesions of normal or yellow coloration and uncertain etiology. In contrast, mucus retention cysts are epithelium-lined cavities originated from a salivary gland. They are also raised, asymptomatic, slow-growing lesions, located on the floor of the mouth, buccal mucosa and lips. This article reports a diagnostic and a surgical treatment of a rare fibrolipoma case associated with a mucus retention cyst located in the palate. PMID- 20027135 TI - A review: oxidative stress in fish induced by pesticides. AB - The knowledge in oxidative stress in fish has a great importance for environmental and aquatic toxicology. Because oxidative stress is evoked by many chemicals including some pesticides, pro-oxidant factors' action in fish organism can be used to assess specific area pollution or world sea pollution. Hepatotoxic effect of DDT may be related with lipid peroxidation. Releasing of reactive oxygen species (ROS) after HCB exposure can be realized via two ways: via the uncoupling of the electron transport chain from monooxygenase activity and via metabolism of HCB major metabolite pentachlorophenol. Chlorothalonil disrupts mitochondrial metabolism due to the impairment of NADPH oxidase function. Activation of spleen macrophages and a decrease of catalase (CAT) activity have been observed after endosulfan exposure. Excessive release of superoxide radicals after etoxazole exposure can cause a decrease of CAT activity and increase phagocytic activity of splenocytes. Anticholinergic activity of organophosphates leads to the accumulation of ROS and resulting lipid peroxidation. Carbaryl induces changes in the content of glutathione and antioxidant enzymes activities. The antioxidant enzymes changes have been observed after actuation of pesticides deltamethrin and cypermethrin. Bipyridyl herbicides are able to form redox cycles and thereby cause oxidative stress. Low concentrations of simazine do not cause oxidative stress in carps during sub-chronic tests while sublethal concentrations of atrazin can induce oxidative stress in bluegill sunfish. Butachlor causes increased activity of superoxide dismutase -catalase system in the kidney. Rotenon can inhibit the electron transport in mitochondria and thereby increase ROS production. Dichloroaniline, the metabolite of diuron, has oxidative effects. Oxidative damage from fenpyroximate actuation is related to the disruption of mitochondrial redox respiratory chain. Low concentration of glyphosate can cause mild oxidative stress. PMID- 20027136 TI - Cyanohepatotoxins influence on the neuroendocrine and immune systems in fish - a short review. AB - Cyanotoxins are the metabolites of cyanobacteria, belonging to different chemical groups and of diverse mechanisms of toxicity. Generally, they are divided into hepatotoxins, neurotoxins and dermatotoxins/irritant toxins. There is a growing evidence, that besides the above mentioned toxicity, exposure to cyanotoxins may also induce other effects, among others the disruption of neuroendocrine and immune systems. The purpose of that paper is to sum up the current information obtained from the literature and from our own studies about the influence of cyanohepatotoxins on neuroendocrine and immune systems of fish. From the presented data it appears, that microcystins, nodularin and cylindrospermopsin, except for their hepatotoxic activity, are potent to exert such effects as HPI axis activation resulting in physiological and behavioural changes, disturbances in thyroid hormones release/metabolism, as well as impairment of immune responses in fish. However the studies in that area are still incomplete and many questions remain to be answered, especially what consequences for fish population health status it brings. PMID- 20027137 TI - The significance of pork as a source of dietary selenium - an evaluation of the situation in the Czech Republic. AB - The aim of this paper is to review current knowledge of the significance of pork as a source of Se for human consumption, and to evaluate the selenium content in pork produced in the Czech Republic. Selenium has an important role in human health. Pork could possibly be an important source of Se for the human diet. The Se content in meat can be increased by a higher dietary intake of selenium in animal feed. The magnitude of this increase is higher when organic Se from Se enriched yeast is ingested. Selenium intoxication in pigs occurs only when very high Se concentrations are used in their diet (more than 5 mg/kg of Se). Organic Se sources produce fewer clinical signs of high toxicity than inorganic sodium selenite. Organic Se does not have a negative effect on meat quality. Altogether, 135 pork samples from 9 different herds in the Czech Republic were collected and analyzed for Se content. The average selenium content found in pork in our study was 87.10 microg/kg. Because the average annual consumption of pork in the Czech Republic is 42.0 kg per person, the annual selenium intake from pork represents 18.2% of the minimum yearly requirement for humans. The results of our study show that pork contributes significantly to the selenium intake of the human population in the Czech Republic. PMID- 20027138 TI - Effects of sesame oil in the model of adjuvant arthritis. AB - OBJECTIVES: The goal of this study was to evaluate the effect of sesame oil on functional damage induced by adjuvant arthritis (AA) and on changes of selected biochemical parameters reflecting oxidative tissue injury. DESIGN: Mycobacterium butyricum in incomplete Freund's adjuvans was intradermally administered to Lewis male rats. Hind paw edema and endothelium-dependent relaxation of the aorta were determined on day 28. Further, plasmatic levels of TBARS, gamma glutamyltransferase (GGT) activity in the joint and spleen tissues, level of protein carbonyls and total antioxidant capacity (TAC) in plasma, as well as activity of the lysosomal enzyme N-acetyl-glucosaminidase (NAGA) in serum were assessed. The effect of sesame oil (SO, 1ml/kg, daily oral administration) was evaluated on day 28. RESULTS: The beneficial effect of sesame oil on markers of oxidative stress accompanying AA was demonstrated by decrease of plasma TBARS and decrease of GGT activity in the joint and spleen tissues. Level of protein carbonyls, TAC in plasma and activity of NAGA in serum and in the kidney were improved, yet not significantly. In the hind paw edema the maximal increase was found on day 28 of AA, and in the same time we observed a significant decrease of aortic endothelium-dependent relaxation. Administration of SO resulted in mild, non-significant decrease of hind paw swelling and in significantly increased acetylcholine-evoked relaxation. CONCLUSION: We conclude that SO has beneficial effects on oxidative stress induced biochemical changes occurring in AA, moreover it improves endothelium-dependent relaxation of the aorta and tends to decrease hind paw edema. PMID- 20027139 TI - Polymorphisms in the adenomatous polyposis coli gene in Slovak families suspected of FAP. AB - OBJECTIVES: Polymorphism in the adenomatous polyposis coli (APC) gene was analyzed in 33 families suspected of familial adenomatous polyposis (FAP) without identified APC gene mutation. Screening of 104 members of mentioned families for polymorphism in the APC gene, was performed using single strand conformation polymorphism (SSCP) and DNA sequencing. RESULTS: Twelve different types of polymorphism were found in the cohort of the families analyzed. Nine polymorphisms were located within exon 15, one within exon 6, one within exon 11 and one within exon 13. Of the 12 polymorphisms, 11 were silent substitution and only one was responsible for the amino acid change - D1822V, which was identified in 60% of the families analyzed. CONCLUSION: The most frequently detected polymorphism D1822V is potentially associated with the risk of colorectal cancer. Three detected polymorphisms - Y486Y, T1493T and S1756S - also seem to be associated with colon cancer risk. All these polymorphisms may be used as markers for diagnosis of colorectal cancer. The importance of other detected polymorphisms remains still unclear, but their involvement is being continuously observed. PMID- 20027140 TI - Conjugated hyperbilirubinaemia as the first manifestation of mevalonic aciduria in a term newborn. AB - OBJECTIVES: To present clinical and laboratory findings in the case of a term newborn with conjugated hyperbilirubinaemia and to stress the importance of differential diagnosis. RESULTS: A term newborn delivered by caesarean section (birth weight 2550 g, birth length 47 cm, value of Apgar score 9/10) with good direct adaptation had on the first day of life increased levels of conjugated bilirubin (23 micromol/l), unconjugated bilirubin (55 micromol/l) and C-reactive protein 39.4 g/l. The diagnosis of mevalonic aciduria was confirmed by urine analysis (mevalonolactone 393 micromol/mmol crea, normal range <2.0 micromol/mmol crea; mevalonic acid 40.5 micromol/mmol crea, normal range <0.04 micromol/mmol crea). CONCLUSION: Mevalonic aciduria can be clinically distinguished based on symptoms of neurological involvement. It can also present itself with hepatosplenomegaly, lymphadenopathy, anaemia, leukocytosis, increased sedimentation rates and levels of C-reactive protein. In cases of conjugated hyperbilirubinaemia of unknown aetiology it is important to exclude mevalonic aciduria by urine investigation for organic acids. PMID- 20027141 TI - Modulation of antioxidative response in the therapy of hypertension and other cardiovascular diseases. AB - OBJECTIVES: This paper reviews and compares major approaches and strategies to modulation of antioxidative response in the therapy of hypertension and cardiovascular diseases. DESIGN: There are two major strategies of modulation of antioxidative response in hypertension and cardiovascular diseases: (i) modulation of NO levels by NOS stimulation, increase of NO bioavailability, administration of NO, and NOS gene incorporation; (ii) scavenging of superoxide and suppression of oxidative stress by activation of antioxidant gene expression or by suppression of selected genes by RNA silencing. These strategies are accomplished by several concepts, including (1) delivery of external agents, (2) antioxidant gene therapy and RNA silencing, and (3) combined therapies and approaches. CONCLUSION: Combined therapies and approches often achieve multiplicative effects and are the most promising attitude in antioxidant oriented therapy of hypertension and cardiovascular diseases. PMID- 20027142 TI - Porcine CYP2A19, CYP2E1 and CYP1A2 forms are responsible for skatole biotransformation in the reconstituted system. AB - OBJECTIVES: To study the contribution of individual purified porcine CYP1A2, 2E1 and 2A19 enzymes to the biotransformation of skatole. METHODS: Individual porcine and human enzymes (CYP1A2, 2E1 or 2A6/19) were used to study their potential involvement in skatole metabolism. Furthermore, the inhibition experiments using specific inhibitors of CYP1A2, 2E1 or 2A6/19, were performed. For determination of skatole biotransformation by individual CYP forms in reconstituted systems, HPLC method with UV detection was used. RESULTS: The data presented in this paper show that porcine and human CYPs are responsible for the formation of indole-3 carbinol and 3-methyloxindole. Whereas in pig CYP2A19 and CYP1A2 seem to be the most important for metabolism of skatole, in man CYP1A2 and CYP2E1 forms are mainly responsible for the production of the metabolites mentioned above. CONCLUSIONS: The porcine and human CYP1A2, 2E1, 2A6/19 forms contribute to formation of 3-methyloxindole and indole-3-carbinol. PMID- 20027143 TI - Influence of probiotics on rat liver biotransformation enzymes. AB - OBJECTIVES: The aim of the study was to find, whether probiotic Escherichia coli Nissle 1917 O6:K5:H1 (EcN) influence the amount and activity of cytochromes P450 (CYP) in rat liver. DESIGN: Live bacterial suspension of EcN was applied to the female Wistar rats in single dose or for 14 days consecutively. The bacterial lipopolysaccharide (LPS) isolated by phenol extraction from the EcN was given to the rats for 14 days as well. Control rats were treated with the saline solution daily for 14 days. Relative amount of CYP2C6, CYP2C9 (corresponding to rat CYP2C11), CYP3A1 and CYP1A2 protein expression in rat liver microsomes was determined by Western blotting. For the determination of six CYP activities (corresponding to human CYP1A2, CYP2A6, CYP2B6, CYP3A4, CYP2C9 and CYP2D6) fluorescence, luminescence or absorbance detection was used. RESULTS: The data presented show that the changes of the total content of the CYP enzymes in rat liver are not significant after administration of the probiotic for 1 or 14 days as well as of the LPS. Western blots revealed a slight increase in CYP2C6 protein expression; level of another rat CYP2C protein (readings with anti-human CYP2C9 antibody corresponding to the rat CYP2C11) as well as of CYP1A2 was elevated after administration of LPS; a small decrease was observed with CYP3A1 protein. Changes in activities of CYP forms are not significant, only the activity of rat CYP2C forms in liver microsomal samples of rats given free LPS appeared to exhibit a small, but significant tendency to increase. CONCLUSION: The results show that the p.o. administration of probiotics to rat does not markedly influence the rat hepatic CYP enzymes. PMID- 20027144 TI - Rat cytochromes P450 oxidize 2-nitrophenol, a human metabolite of carcinogenic 2 nitroanisole. AB - OBJECTIVES: 2-Nitrophenol (2-NP) is the major detoxification metabolite of an important industrial pollutant and a potent carcinogen, 2-nitroanisole (2-NA). Characterization of the products of 2-NP metabolism by rat hepatic microsomes containing cytochromes P450 (CYPs) and identification of the major CYP enzymes participating in this process are aims of this study. METHODS: HPLC with UV detection was employed for the separation and characterization of 2-NP metabolites. Inducers and inhibitors of CYPs and rat recombinant CYPs were used to characterize the enzymes participating in 2-NP oxidation. RESULTS: Rat hepatic microsomes oxidize 2-NP to its hydroxylated metabolite, 2,5-dihydroxynitrobenzene (2,5-DNB). No nitroreductive metabolism leading to the formation of o-aminophenol was evident when using rat hepatic microsomes. Selective CYP inhibitors and hepatic microsomes of rats pre-treated with specific CYP inducers were used to characterize CYPs oxidizing 2-NP in rat livers. Based on these studies, we attribute most of 2-NP oxidation in rat liver to CYP2E1 and 3A, followed by CYP2D and 2C. Among recombinant rat CYP enzymes tested in this study, CYP2E1 and 2C11 were the most effective enzymes oxidizing 2-NP. Oxidation of 2-NP by rat CYP2E1 exhibits the Michaelis-Menten kinetics, having the Km value of 0.35 mM. CONCLUSION: The results found in this study, the first report on the metabolism of 2-NP by rat hepatic microsomes and rat CYP enzymes, demonstrate that CYP2E1 is the major enzyme oxidizing this compound in rat liver. PMID- 20027145 TI - Oxidation of 3-aminobenzanthrone, a human metabolite of carcinogenic environmental pollutant 3-nitrobenzanthrone, by cytochromes P450 - similarity between human and rat enzymes. AB - OBJECTIVES: 3-Aminobenzanthrone (3-ABA) is the main human metabolite of carcinogenic environmental pollutant 3-nitrobenzanthrone (3-NBA). Understanding which cytochrome P450 (CYP) enzymes are involved in metabolism of this toxicant is important in the assessment of individual susceptibility. Characterization of 3-ABA metabolites formed by rat hepatic microsomes containing cytochromes P450 (CYPs) and identification of the major rat and human CYPs participating in this process are aims of this study. METHODS: HPLC with UV detection was employed for the separation and characterization of 3-ABA metabolites. Inducers and inhibitors of CYPs and rat and human recombinant CYPs were used to characterize the enzymes participating in 3-ABA oxidation. RESULTS: Selective CYP inhibitors and hepatic microsomes of rats pre-treated with specific CYP inducers were used to characterize rat liver CYPs metabolizing 3-ABA (measured as consumption of 3 ABA). Kinetics of these reactions catalyzed by rat hepatic microsomes was also evaluated. Based on these studies, we attribute most of 3-ABA metabolism in rat liver to CYP1A and 3A. Among recombinant rat and human CYP enzymes tested in this study, rat CYP3A2 and human CYP3A4/5, followed by CYP1A1 of both organisms were the most effective enzymes converting 3-ABA. Rat hepatic CYP enzymes oxidize 3 ABA up to three metabolites. Two of them were identified to be the products formed by oxidation of 3-ABA on its amino group back to the parent compound from which 3-ABA is generated in organisms, 3-NBA. Namely, N-hydroxylation metabolite, N-hydroxy-3-ABA and 3-NBA were identified to be these 3-ABA oxidation products. These metabolites are formed by CYPs of a 1A subfamily. Another 3-ABA metabolite, whose structure remains to be characterized, is generated not only by CYP1A but also by other CYP enzymes, predominantly by CYPs of a 3A subfamily. CONCLUSION: The results found in this study, the first report on the metabolism of 3-ABA by human and rat CYPs, clearly demonstrate that CYPs of 3A and 1A subfamilies are the major enzymes metabolizing 3-ABA. PMID- 20027146 TI - Cytotoxicity of and DNA adduct formation by ellipticine in human U87MG glioblastoma cancer cells. AB - OBJECTIVES: Ellipticine is a potent antineoplastic agent exhibiting multiple mechanisms of action with promising brain tumor specificity. This anticancer agent should be considered a pro-drug, whose pharmacological efficiency and/or genotoxic side effects are dependent on its cytochrome P450 (CYP) - and/or peroxidase-mediated activation to species forming covalent DNA adducts. Ellipticine can also act as an inhibitor or inducer of biotransformation enzymes, thereby modulating its own metabolism leading to its genotoxic and pharmacological effects. The toxicity of ellipticine to U87MG glioblastoma cells and mechanisms of its action to these cells are aims of this study. METHODS: Ellipticine metabolites formed in U87MG cells were analyzed using HPLC. Covalent DNA modifications by ellipticine were detected by 32P-postlabeling. CYP enzyme expression was examined by QPCR and Western blot. RESULTS: U87MG glioblastoma cell proliferation was efficiently inhibited by ellipticine. This effect might be associated with formation of two covalent ellipticine-derived DNA adducts, identical to those formed by 13-hydroxy- and 12-hydroxyellipticine, the ellipticine metabolites generated by CYP1A1, 1B1 and 3A4, lactoperoxidase and cyclooxygenase 1, the enzymes expressed in U87MG cells. Moreover, by inducing CYP1B1, 3A4 and 1A1 enzymes in U87MG cells, ellipticine increases its own enzymatic activation, thereby enhancing its own genotoxic and pharmacological potential in these cells. Ellipticine concentration used for U87MG cell treatment is extremely important for its pharmacological effects, as its metabolite profiles differed substantially predicting ellipticine to be either detoxified or activated. CONCLUSION: The results found in this study are the first report showing cytotoxicity and DNA adduct formation by ellipticine in glioblastomas. PMID- 20027147 TI - Modulation of cytochrome P450 enzyme system by selected flavonoids. AB - OBJECTIVES: The aim of this study was to assess the effect of various flavonoids on the NADPH:cytochrome P450 oxidoreductase (CYPOR) activity in respect of the reduction of different electron acceptors as well as to study an impact of flavonoids on monooxygenation of a model substrate of cytochrome P450 (CYP). DESIGN: The modulation of CYPOR activity was determined spectrophotometrically based on the time course of the reduction of different electron acceptors. The CYP reduction was monitored via its complex formation with CO, having pronounced the absorption maximum at 450 nm. Finally, effect of CYPOR stimulation by 7,8benzoflavone (ANF) on 7pentoxyresorufin Odepentylation was assayed in the microsomal monooxygenation system using the fluorimetric detection of formed resorufin. RESULTS: The stimulation of CYPOR activity via ANF was found to be associated with following electron acceptors: cytochrome c, potassium ferricyanide, cytochrome b5, but not with CYP. Surprisingly, 5,6benzoflavone, a position isomer of ANF, was ineffective in the CYPOR stimulation as well as the other flavonoids tested. In microsomal preparations, ANF did not markedly enhance the reaction rate of monooxygenation of CYP2B4 model substrate. CONCLUSION: Our results document that among all of the tested flavonoids only ANF is able to stimulate CYPOR activity, however, the ANF-mediated stimulation of CYPOR has no impact on the oxidative metabolism catalyzed by CYP system. PMID- 20027148 TI - Preparation of apo-cytochrome b5 utilizing heme transfer to apo-myoglobin. AB - OBJECTIVES: Cytochrome b5 (cyt b5), a component of endoplasmic reticulum membrane, plays a role in modulation of activity of several cytochromes P450 (CYP). To elucidate the mechanism of such modulations it is necessary to evaluate not only the effect of native cyt b5, but also that of apo-cyt b5. To prepare apo cyt b5, heme transfer from native cyt b5 to a protein with higher affinity toward the heme, the horse heart apo-myoglobin, was utilized. METHODS: Butanone extraction was employed to prepare apo-myoglobin. Apo-cyt b5 was separated from myoglobin by chromatography on DEAE-Sepharose. Mass spectrometry was utilized to characterize proteins eluted from DEAE- Sepharose. RESULTS: The prepared apo myoglobin was incubated with the cyt b5 at pH 4.2 that is the optimal pH for heme transfer from cyt b5 into apo-myoglobin. The apo-cyt b5 protein was separated from myoglobin present in the reaction mixture by chromatography on a column of DEAE-Sepharose. Using such a procedure, 16% yield of apo-cyt b5 that did not contain any heme in its molecule was obtained from the native rabbit cyt b5. Oxidized and reduced forms of the apo-b5 reconstituted with heme exhibit the same absorbance spectra as native cyt b5. The prepared apo-cyt b5 reconstituted with heme can receive electrons from NADPH:CYP reductase. CONCLUSION: A biologically active apo-cyt b5 was prepared using transfer of heme from cyt b5 to horse heart apo-myoglobin by the procedure described here. PMID- 20027149 TI - Isolation and partial characterization of catechol 1,2-dioxygenase of phenol degrading yeast Candida tropicalis. AB - OBJECTIVES: Candida tropicalis yeast is a microorganism that possesses high tolerance for phenol and shows strong phenol degrading activity. This yeast is capable of utilizing phenol as the sole carbon and energy source. While the enzyme participating on the first step of phenol biodegradation, NADPH-dependent phenol hydroxylase, has already been characterized, information on the enzyme participating in the second step of its degradation, catechol 1,2-dioxygenase, is scarce. The development of the procedure suitable for catechol 1,2-dioxygenase isolation and partial characterization of this enzyme are the aims of this study. METHODS: Combination of chromatography on DEAE-Sepharose and gel-permeation chromatography on Sephadex G-100 was used for isolation of cytosolic catechol 1,2 dioxygenase from C. tropicalis yeast. The sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and gel chromatography on Sephadex G-100 were used to evaluate the molecular mass of the enzyme. The enzyme activity was followed by HPLC (catechol consumption and/or cis,cis-muconic acid formation). RESULTS: Using the isolation procedure consisting of chromatography and re-chromatography on a column of DEAE-Sepharose and gel filtration on Sephadex G-100, catechol 1,2 dioxygenase was purified from C. tropicalis cytosol to homogeneity. Catechol 1,2 dioxygenase was found to be a homodimer with a subunit molecular mass of 30000 +/ 5000. The enzyme oxidized catechol producing cis,cis-muconic acid. The optimal temperature and pH were 30 degrees C and 7.7, respectively. CONCLUSIONS: The data are the first report showing the isolation of eukaryotic catechol 1,2-dioxygenase from C. tropicalis to homogeneity and its partial characterization. PMID- 20027150 TI - Intersexual differences in inhibitory influence of trans-resveratrol on activity of cytochrome P450 2D2 in rats. AB - OBJECTIVES: Differences in the metabolism between males and females have been seen over time. Hormonal regulation of cytochrome P450 activity is understood to be involved. Trans-resveratrol (RES) is an estrogenically active plant polyphenol with many protective biological activities including neuroprotection. The present report studied the influence of sex and RES on variances in rat's cytochrome P450 2D2 hepatic metabolic activity. METHODS AND DESIGN: Isolated perfused rat liver was used for determination of cytochrome P450 2D2 activity. Wistar albino rats of both sexes were treated with RES at the dose of 5 mg/kg/day for 10 days prior to liver isolation. Levels of marker substance dextromethorphan (DEM) and its 2D2 specific metabolite dextrorphan (DEX) were measured during perfusion. The metabolic ratios (DEM/DEX) and the levels of DEM and DEX in perfusate were compared. RESULTS: In the controls, the activity of CYP2D2 was found to be higher in male rats compared to females. RES produced inhibition of CYP2D2, expressed by significant changes of both DEM and DEX levels in males and significant increase of only DEM levels in females. There were no gender changes in DEX levels in RES treated animals whilst DEM levels were significantly increased during the whole perfusion in females. CONCLUSION: The results confirmed gender differences in the metabolic activity of CYP450 2D2 with a higher rate in male rats. RES acted as an inhibitor, however again with greater impact in males than in females. This metabolic divergence could be a cause for different sensitivity or even toxicity of drugs metabolized by the CYP450 2D2. PMID- 20027151 TI - Evaluation of the activity of P450 enzymes in rats: use of the single marker or combined drug administration. AB - OBJECTIVES: A "cocktail" of several substrates is frequently used to assess metabolic activity of multiple cytochrome P450 enzymes in one session. Some interactions among substrates can appear and may influence the rate of biotransformation of other ones. Our current work was aimed on the influence of tolbutamide on cytochrome P450-mediated metabolism of phenacetin and vice versa. DESIGN: In the presented work, the biotransformation rates of phenacetin and tolbutamide (markers of rat CYP1A2 and CYP2C6/11 metabolic activities, respectively) administered either separately or both simultaneously were compared. The model of isolated perfused rat liver was used. RESULTS: Phenacetin had no significant effect on tolbutamide hydroxylation. Tolbutamide addition to the perfusion medium significantly increased the rate of O-deethylation of phenacetin. CONCLUSION: Some differences in the rate of P450-mediated metabolism can be observed when comparing assessment using combination of two model substrates with the common way (single marker administration). Due to these differences, results obtained by the mentioned methodologies might not be fully comparable. PMID- 20027152 TI - Type and distribution of indomethacin-induced lesions in the gastrointestinal tract of rat. AB - OBJECTIVES: The therapy with non-steroidal anti-inflammatory drugs (e.g. indomethacin) is often accompanied with adverse effects in gastrointestinal tract. Aim of this experimental study was to define the time range of the creation of indomethacin-induced gastrointestinal lesions in rat (for prospective study of potential probiotic therapy). The paper follows our previous experiments where the different gastrointestinal lesions were described in the pig (Kvetina et al. 2008) METHODS: Indomethacin (25mg/kg) was administered orally by a single application to rat (Wistar Han II, 200-250g). Six, 24, 48 and 72 hours after the indomethacin administration all parts of the gastrointestinal tract of six rats in each time interval were macroscopically and histologically examined. RESULTS AND CONCLUSION: The gradual development of lesions was observed 6 hours in stomach and 24-72 hours in the intestine after the indomethacin administration. Not only the gradual development of pathophysiological alterations was observed but also the reparative phase (in stomach). 24 hours seem to be advisable time suitable for the evaluation of the probiotics effect as a potential therapy) on the indomethacin-induced gastrointestinal lesions in rats. Sensitivity of the gastrointestinal tract to the pathological lesions development seems to be higher in rats in comparison to findings described in our previous experiments in pig (Kvetina et al. 2008). This adverts to interspecies differences in the manifestation and in the dynamics of the development of gastrointestinal lesions. PMID- 20027153 TI - Caco-2 cells and Biopharmaceutics Classification System (BCS) for prediction of transepithelial transport of xenobiotics (model drug: caffeine). AB - OBJECTIVES: The Caco-2 cell monolayer model is widely used as a standard screening tool for studying the mechanisms of cellular drug transport. Caffeine was chosen as a model drug and is supposed to be class I of the Biopharmaceutics Classification System (BCS). Our study was conducted 1) to characterize the mechanisms of caffeine transport across the intestinal barrier, 2) to classify caffeine according to BCS, 3) to predict drugs intestinal absorption in humans. METHODS: Caffeine transport (0.1, 0.3, 1 and 10 mmol/l) was studied in Caco-2 cell monolayer in apical to basolateral (AP-BL) and basolateral to apical (BL-AP) direction, under iso-pH 7.4 and pH-gradient (6/7.4) conditions. The relative contribution of the paracellular route was estimated using Ca2+- free transport medium (opening tight junctions). RESULTS: The caffeine transport was linear with time, transport direction and pH independent, displaying non-saturable (first order) kinetics, with high permeability coefficient (Papp): in AP-BL direction Papp = 46.3-53.5 x 10-6 cm/s; in BL-AP direction Papp = 45.6-49.4 x 10-6 cm/s. Thus, the transport seems to be transcellular mediated by passive diffusion. Using Ca2+- free transport medium tight junctions were opened (confirmed by increased Papp of mannitol) but the caffeine Papp was not changed. Thus, the paracellular route is only a minor way of caffeine transport. CONCLUSION: High solubility and high permeability of caffeine rank it among class I of BCS and well absorbed compounds. PMID- 20027154 TI - The effect of different molecular weight hyaluronan on macrophage physiology. AB - OBJECTIVES: Hyaluronan, a linear glycosaminoglycan, is an abundant component of extracellular matrix. In its native form, the high-molar-mass hyaluronan polymers have an array of structural and regulatory, mainly anti-inflammatory and anti angiogenic, functions. In contradiction, the biological effects of fragmented low molecular weight hyaluronan are suggested to be pro-angiogenic and pro inflammatory. METHODS: The effects of highly purified pharmacological grade hyaluronan of defined molecular weights 11, 52, 87, 250 and 970 kilodaltons were tested on mouse macrophage cell lines RAW 264.7 and MHS. The surface expression of CD44 and Toll-like receptor 2, surface receptors for hyaluronan, was determined by flow cytometry. Activation of macrophages was determined based on nitric oxide and tumour necrosis factor alpha production, inducible nitric oxide synthase expression, and the activation of the nuclear factor kappa B transcriptional factor. RESULTS: Both macrophage cell lines expressed CD44 and Toll-like receptor 2, which were significantly increased by the pre-treatment of macrophages with bacterial lipopolysaccharide. Hyaluronan of any molecular weight did not activate production of nitric oxide or tumour necrosis factor alpha in any mouse macrophage cell lines. Correspondingly, hyaluronan of any tested molecular weight did not stimulate nuclear factor kappa B activation. Similarly, hyaluronan of any molecular weight neither exerted stimulatory nor inhibitory effects on macrophages pre-treated by lipopolysaccharide. CONCLUSION: Interestingly, the data does not support the current view of low molecular weight hyaluronan as a pro-inflammatory mediator for macrophages. Further studies are necessary to clarify the effects of different molecular weight hyaluronan on phagocytes. PMID- 20027155 TI - The effect of uric acid on homocysteine-induced endothelial dysfunction in bovine aortic endothelial cells. AB - OBJECTIVES: Elevated plasma uric acid indicates an increased risk of cardiovascular diseases associated with endothelial dysfunction. However, the role of uric acid in the pathogenesis of endothelial dysfunction is still a matter of debate. It is not clear whether uric acid is a real causative risk factor, an inert marker, or even a protective molecule with respect to its antioxidant properties. We have studied the effect of uric acid on intact endothelial cells as well as cells with homocysteine-induced endothelial dysfunction. DESIGN: Bovine aortic endothelial cells were treated with uric acid (100 - 600 muM) and homocysteine (100 muM) or with uric acid only. After 24 hours, the cells were stimulated with 1 mug/ml of calcium ionophore A23187, and nitric oxide (NO) production was measured electrochemically with the use of a NO sensitive microelectrode. The expression of endothelial nitric oxide synthase (eNOS) and eNOS phosphorylation at Ser1179 was estimated with the use of Western blotting. Interaction between NO and uric acid was measured with a NO electrode. Superoxide generation was measured with the use of the fluorescence dye MitoSox Red. RESULTS: Homocysteine strongly diminished A23187-induced NO release. 100 muM uric acid slightly restored NO production; higher concentrations were ineffective. Interestingly, a dose-dependent decrease of NO release was observed in the cells treated only with uric acid. Uric acid did not scavenge NO and did not change eNOS protein expression or phosphorylation at Ser1179, but dose dependently increased superoxide production in A23187-stimulated cells. CONCLUSION: In conclusion, uric acid decreased NO bioavailability and enhanced superoxide generation in A23187-stimulated bovine aortic endothelial cells. PMID- 20027156 TI - Increased markers of oxidative stress in plasma of patients with chronic pancreatitis. AB - OBJECTIVES: Chronic pancreatitis (CP) is a heterogeneous disease defined as chronic inflammatory changes of the pancreatic tissue caused by variety of aetiologies. Oxidative stress accompanying the inflammatory processes has been suggested as an important factor contributing to CP development. The aim of this study was to determine levels of lipid peroxidation products malondialdehyde (MDA) and 4-hydroxynonenal (4-HNE), together with nitrites and the total antioxidant capacity in the plasma of patients with CP and control subjects. DESIGN: One hundred and five patients with chronic pancreatitis and twenty seven healthy controls were included into this study. Levels of MDA and 4-HNE were analyzed using high-performance liquid chromatography. The total antioxidant capacity of plasma against peroxyl radicals was evaluated using chemiluminescent determination. Nitrites were determined using Griess reaction. Biochemical and haematological parameters were measured by standard methods. RESULTS: The plasma levels of both MDA and 4-HNE, together with the plasma levels of nitrites, were significantly higher in CP patients, compared to healthy controls. The total antioxidant capacity did not differ significantly. Biochemical parameters were in the normal range. The MDA and 4-HNE levels correlated positively with the levels of high-density lipoprotein cholesterol. Nitrite levels correlated positively with C-reactive protein, total white blood cells, and triglycerides. CONCLUSION: The significantly increased plasma levels of MDA, 4-HNE, and nitrites indicate that oxidative stress is present in patients with CP and that it may play a role in initiation and maintenance of inflammation within the pancreatic tissue in CP patients. PMID- 20027157 TI - The comparison of impedance-based method of cell proliferation monitoring with commonly used metabolic-based techniques. AB - OBJECTIVES: Determination of cell numbers is a crucial step in studies focused on cytokinetics and cell toxicity. The impedance-based analysis employing electronic sensor array system xCELLigence System allowing label-free dynamic monitoring of relative viable adherent cell amounts was compared with the most utilized methods for relative quantification of viable cell numbers based on a determination of cellular metabolism. DESIGN: Colorimetric assay based on reduction of tetrazolium salt (MTT) by mitochondrial enzymes and chemiluminiscent assay based on intracellular adenosine triphosphate (ATP) determination were compared with the impedance-based system. Cell morphology was compared by microscopic evaluation. Normal human epidermal keratinocytes (NHEK) and normal human dermal fibroblasts (NHDF), together with 3T3 mouse fibroblast and HaCaT keratinocyte cell lines were employed. RESULTS: The progress of cell growth curves obtained by different methods during 72 hours reflected cell type and cell seeding densities. The impedance-based method was found to be applicable for the determination of the cell proliferation of 3T3 fibroblasts, HaCaT and NHDF, since the comparison of this method with ATP and MTT determinations showed a comparable results. In contrast, the proliferation of NHEK measured by the impedance-based method did not correlate with other methodological approaches. This could be accounted to the specific morphological appearance of these cells. CONCLUSION: The study shows the impedance-based detection of viable adherent cells is a valuable approach for cytokinetics and pharmacological studies. However, the specific morphological characteristics of cell lines have to be considered employing this method for determination of cell proliferation without using other reference methods. PMID- 20027158 TI - Interactions of oxidatively modified calf skin collagen with platelets and phagocytes. AB - OBJECTIVES: The effects of non-modified and oxidatively modified calf skin collagen type I on platelet aggregation and the oxidative burst of phagocytes were examined in the framework of a general hypothesis that collagen, platelets and phagocytes cooperate to modulate the oxidative burst of phagocytes and the extent of oxidative stress. MATERIALS AND METHODS: Calf skin collagen type I was subjected to oxidative modification by hydrogen peroxide or hydroxyl radical. Thermal denaturation of collagen was performed in a spectrophotometer equipped with a temperature gradient device. The aggregation of isolated human platelets obtained after differential centrifugation was measured using a dual-channel aggregometer. The production of reactive oxygen species by human whole blood phagocytes was evaluated by luminol-enhanced chemiluminescence. RESULTS: Oxidative modification of collagen samples was characterized by a decrease in denaturation transition temperature. Oxidatively modified samples showed a modified SDS-PAGE pattern, evidencing a significant destruction of the collagen. All oxidatively modified collagen samples, independent of the oxidation treatment applied, lost their platelet-aggregating and phagocyte oxidative burst-inducing activity. CONCLUSION: The results suggest that reactive oxygen species were able to modify collagen. On the other hand, oxidatively modified collagen lost its activating properties towards platelets and phagocytes. PMID- 20027159 TI - H1-antihistamines and oxidative burst of professional phagocytes. AB - OBJECTIVES: We analysed and compared the effect of five H1-antihistamines on stimulated oxidative burst at extra- and intracellular level of isolated and stimulated human polymorphonuclear leukocytes. DESIGN: Oxidative burst of isolated human neutrophils was studied by means of luminol and isoluminol enhanced chemiluminescence. RESULTS: The following rank order of potency for H1 antihistamines to decrease chemiluminescence was evaluated extracellularly: dithiaden> loratadine> chlorpheniramine> brompheniramine> pheniramine and at intracellular site: loratadine> dithiaden. CONCLUSION: H1-antihistamines differ substantially according to their chemical structure in suppressing oxidative burst both at extra- and intracellular site of isolated stimulated human neutrophils. PMID- 20027160 TI - Importance of thiopurine S-Methyltransferase gene polymorphisms for prediction of azathioprine toxicity. AB - OBJECTIVES: Our study aims to find the relationship between metabolic enzyme thiopurine S-methyltransferase (TPMT) gene polymorphisms and clinical output of the therapy with azathioprine. We focused on patients who experienced leucopenia caused by high blood levels of active azathioprine metabolites. DESIGN: Our group consists of 87 patients who have been treated by azathioprine. 21 individuals experienced leucopenia during treatment with standard dose of azathioprine. We have used PCR-REA and "real-time" PCR methods for genotype detection G238C, G460G and A719G substitutions in TPMT gene. RESULTS: We have found statistical association between the presence of non-standard TPMT alleles and adverse event associated with azathioprine treatment - leucopenia (p=0.0033). CONCLUSION: Our results confirm that TPMT genotyping prior to the treatment with azathioprine could predict patients with genetic predisposition for serious leucopenia and seems to be a useful genetic marker for individualisation of the therapy. PMID- 20027161 TI - Evaluation of selenium status in pure bred duroc sows and their progeny. AB - OBJECTIVES: The aim of this trial was to determine selenium status in pure bred duroc sows and their progeny and to compare it to Czech Large White x Landrace breed. DESIGN: The pregnant duroc sows (n=12) and pregnant Czech Large White x Landrace sows (n=12) were fed identical diets supplemented with sodium selenite. RESULTS: During lactation significantly higher serum Se concentrations (p<0.001) were found in duroc piglets. Also significantly higher serum GSH-Px activities (p<0.001) were found in duroc sows and piglets. No differences in concentrations of Se in colostrums and milk were found between the two breeds. CONCLUSION: Our findings support the hypothesis that there may be breed differences in indices of selenium status in pigs. PMID- 20027162 TI - Modulating effect of flavonoids and their derivatives on sarcoplasmic reticulum Ca2+-ATPase oxidized by hypochloric acid and peroxynitrite. AB - OBJECTIVES: Effect of rutin and its lipophilic derivatives on Ca2+-ATPase of sarcoplasmic reticulum (SERCA) oxidized by hypochloric acid and peroxynitrite was investigated to examine the role of flavonoids in SERCA activity modulation. METHODS: Ca2+-ATPase activity was measured spectrophotometrically at 37 degrees C using NADH-coupled enzyme pyruvate kinase/lactate dehydrogenase assay. SERCA was oxidized by HOCl (3 min) or ONOO- (30 s) after previous treatment with flavonoids (2 min) at 37 degrees C. Lipophilic rutin derivatives were prepared by lipase catalyzed esterification of flavonoids with fatty acids. RESULTS: Both hypochloric acid (HOCl) and peroxynitrite (ONOO-) decreased ATPase activity concentration-dependently with IC50 of 50+/-10 micromol/l and 150+/-15 micromol/l, respectively. Rutin was found to have a protective effect on SERCA activity in both oxidation systems in the concentration range 5 - 250 micromol/l. Lipophilic rutin derivatives (rutin oleate, rutin linoleate, rutin linolenate) exerted inhibitory effect on ATPase activity both in the presence and absence of oxidants. CONCLUSION: The results suggest that selective lipophilization of the flavonoid skeleton may represent a useful tool for SERCA activity modulation. PMID- 20027163 TI - Efficacy of structural homoloques and isomers of pralidoxime in reactivation of immobilised acetylcholinesterase inhibited with sarin, cyclosarin and soman. AB - OBJECTIVES: Quantification of efficacy of monopyridinium isomers and homologs derived from clinically used Pralidoxime within reactivation of acetylcholinesterase inhibited with organophosphorus nerve agents. METHODS: This work uses the colorimetric biosensor called Detehit - cotton cloth with immobilized enzyme acetylcholinesterase. Biosensor is based on the modificated Ellman's method. RESULTS: The highest reactivation was observed with sarin inhibited acetylcholinesterase. Substantially lower reactivation was found with the cyclosarin-inhibited enzyme whereas AChE, inhibited by soman could not be effectively reactivated under the given conditions (enzyme inhibition for 2 minutes and subsequent treatment with the reactivator for 15 minutes). CONCLUSION: Our work gives comparison of efficacy of reactivators in dependence on the length of alkylene chain and position of aldoxime functional group. Evaluation of effectivity of aldoxime reactivators is provided by simple means. The method allows rapid in vitro evaluation of the reactivators without being disturbed by excess of the organophosphate or reactivator. PMID- 20027164 TI - Fish oil and cod liver as safe and healthy food supplements. AB - OBJECTIVES: The aim of this study is to determine the content of mercury, methylmercury and persistent organic pollutants in the capsules filled with fish oil from marine fish and in the canned cod liver and find out, whether analyse products are safe and suitable for human consumption. METHODS: Total mercury was determined by cold vapour atomic absorption spectroscopy on an AMA-254 (Altec Ltd., Czech Republic) single-purpose mercury analyzer. Methylmercury in the cod liver in the form of chlorid methylmercury was determined by gas chromatography. Seven indicator congeners of PCB (PCB 28, 52, 101, 118, 138, 153, 180), HCH, HCB, DDT and its degradation products DDE and DDD, were determined by gas chromatography-mass spectrometry. RESULTS: In capsules filled with fish oil (n=19) total Hg levels were in the range of 0.013 to 2.03 ng/g. All the capsule oil samples analyzed for MeHg were below the detection limit. The highest concentration of total Hg was found in cod liver - can A (0.223 ng/g). The maximum percentage of MeHg in total Hg concentration was found in a liver sample can B. The values of alpha-, beta-, gamma-, delta- HCH and of HCB were found below the detection limits in all the capsule samples analyzed. In eight capsules, the presence of highly lipophilic PCB congeners was demonstrated. DDT and its important DDE metabolite were found in ten samples. In canned cod liver the highest concentration of all PCB congeners was demonstrated in can D. The lowest congener concentrations, however, were found in smoked cod liver - can C. DDT with its main metabolite DDE was detected in can C. No additional DDT and DDD persistent pollutants were detected. CONCLUSIONS: The consumption of fish oil in capsules, and canned cod liver is safe and healthy and should be encouraged. PMID- 20027165 TI - Effect of naturally mouldy wheat or fungi administration on metallothioneins level in brain tissues of rats. AB - OBJECTIVE: The aim of this study is to determine level of metallothioneins (MTs) in brain tissues of rats administered by feed mixtures with different content of mouldy wheat or fungi. DESIGN: Selected male laboratory rats of Wistar albino at age of 28 days were used in our experiments. The rats were administered by feed mixtures with different content of vitamins, naturally mouldy wheat or fungi for 28 days. At the very end of the experiment, the animals were put to death and brains were sampled. MT level was determined by differential pulse voltammetry Brdicka reaction. RESULTS: We found that MTs' level in brain tissues from rats administered by standard feed mixtures was significantly higher compared to the level of MTs in rats supplemented by vitamins. Further we studied the effect of supplementation of naturally mouldy wheat on MTs level in rats. In mouldy wheat we detected the presence of following fungi species: Mucor spp., Absidia spp., Penicillium spp., Aspergillus spp. and Fusarium spp. Moreover we also identified and quantified following mycotoxins - deoxynivalenol, zearalenone, T2-toxin and aflatoxins. Level of MTs determined in rats treated with 33 or 66% of mouldy wheat was significantly lower compared to control ones. On the other hand rats treated with 100% of mouldy wheat had less MTs but not significantly. Supplementation of vitamins to rats fed by mouldy wheat had adverse effect on MTs level compared to rats with no other supplementation by vitamins. Moreover vitamins supplementation has no effect on MTs level in brain tissues of rats treated or non-treated with Ganoderma lucidum L. CONCLUSION: Both mycotoxins and vitamins have considerable effect on level of MTs in brain tissues. It can be assumed that the administered substances markedly influence redox metabolism, which could negatively influence numerous biochemical pathways including those closely related with MTs. PMID- 20027166 TI - Effect of metals, with special attention of Cd, content of the Svitava and Svratka rivers on levels of thiol compounds in fish liver and their use as biochemical markers. AB - OBJECTIVES: The aim of this study was to assess the effect of metals, with special attention of cadmium (Cd), content of the Svitava and Svratka rivers (Czech Republic) on levels of thiol compounds such as metallothioneins (MT), glutathione (GSH/GSSG), and cysteine (Cys) concentrations in chub (Leuciscus cephalus L.) liver and their use as biomarkers. DESIGN: Levels of thiol compounds in fish liver were determined by the differential pulse voltammetry Brdicka reaction. Cadmium content in L. cephalus liver was measured by atomic absorption spectometry (AAS) with electrothermal atomization. The contents of thiols and Cd were correlated with the Cd and metals content of river bottom sediments. RESULTS: Results were inconsistent with those of experimental studies. Correlations between liver Cd content and MT, Cys, GSSG were not significant in any locality (p > 0.05). Cd liver content was negatively (p< 0.05) correlated with GSH at one (Zidlochovice) of the seven tested sites. GSH and GSSG correlated negatively in five localities (p > 0.05). Correlations between Cd in sediment and Cys and GSSG (p > 0.05), and MT was negative (p< 0.05). The relationship between sum of metals value and MT was negative (p< 0.05), similarly with GSSG (p > 0.05). CONCLUSION: Many factors influence content of MT, GSH, and GSSG, and data for Cd were equivocal. Monitoring the aquatic ecosystem to identify metal content using thiol compounds in fish liver as biomarkers is complex. More relevant data including laboratory experiments will be required to assess most of the factors influencing thiols and their relevance to Cd contamination. PMID- 20027167 TI - Using human hair as an indicator for exposure to mercury. AB - OBJECTIVES: Exposure to mercury, a risk factor for neuro-developmental toxicity, was evaluated in the Czech Republic by performing mercury determination using human hair as an indicator. METHODS: Hair samples from Czechs (n=311; 2-66 years old) were analyzed for mercury content. Total mercury was analysed by mercury analyzer AMA 254. RESULTS: The highest total mercury content found in sampled hair was 3.55 microg/g and the lowest content was 0.015 microg/g. No correlation was found between the mercury levels in the sampled hair and the subject's age, gender, and the amount of amalgam fillings. A total of 38 hair samples were analyzed for methylmercury content. CONCLUSION: The results show a positive correlation between the total mercury content in human hair and the consumption of marine and freshwater fish. Hair are a very good indicator of fish consumption. PMID- 20027168 TI - Human lung cancer and hexavalent chromium exposure. AB - OBJECTIVE: Workers chronically exposed to hexavalent chromium have higher incidence of lung cancer. Our study investigates incidence of lung cancer types, age at onset of the disease and survival time among chromium exposed workers (smelters, tapers, crane operators) in comparison to non-exposed persons. METHODS: 64 chromium exposed workers and 104 male controls with diagnosed lung cancer were analysed. The average exposure time among workers was 16.71 +/- 10.02 (S.D.) years (range 1- 41 years). RESULTS: Chromium exposure significantly decreases the age at the onset of the disease by 3.51 years (62.20 +/- 9.08 years in exposed group and 65.71 +/- 10.50 years in control; P=0.018). Small cell lung carcinoma (SCLC) forms 25.0 % of all cases in chromium exposed workers and 16.34% in non exposed individuals. No correlation was found between the age at the diseases onset and time of exposure. The mean survival time in exposed group was 9.03 +/- 12.73 month, in control 12.14 +/- 21.94 month, but this difference was not significant (P=0.473). CONCLUSION: Occupational exposure to chromium was identified as an important risk factor of lung cancer, decreasing the age at the diseases onset. Higher percentage of SCLC was found in chromium exposed individuals. PMID- 20027169 TI - Assessment of low-molecular-weight antioxidants in Francisella tularensis infected hosts: comparison of two rodents with different susceptibility to tularemia. AB - OBJECTIVES: Bacterium Francisella tularensis is the causative agent of tularemia disease. It is a zoonosis accompanied with high mortality when untreated. Small rodents and hares, in particular, are natural reservoirs of tularemia. Despite physiological similarity of common hosts, tularemia exerts different mortality rates. The pathogenesis of tularemia is still not fully understood. The main pathway is associated with proliferation in macrophages after activation by reactive oxygen species in phagosomes. DESIGN: A fully virulent strain of F. tularensis subsb. holarctica was used for infection of laboratory BALB/c mice (Mus musculus) and common voles (Microtus arvalis) representing murine and microtine species. The total level of low-molecular- weight antioxidants (LMWA) in plasma was assayed by cyclic voltammetry. RESULTS: It was found that common voles are more resistant to tularemia progression when compared to mice. When LMWA assayed, surprising changes in LMWA levels were found. Both mice and common voles were infected with high dose resulting in overall mortality. While there was a quick depletion of LMWA in plasma in mice, common voles were even able to increase LMWA. CONCLUSION: It seems that LMWA play an important role in the organism s protection during tularemia. The ability to compensate the LMWA losses and increase levels of antioxidants in common voles is probably responsible for its lower susceptibility to tularemia. PMID- 20027170 TI - Polyaluminium chloride (PAX-18) - acute toxicity and toxicity for early development stages of common carp (Cyprinus carpio). AB - OBJECTIVES: The aim of this study is to assess the toxicity of PAX-18 in different developmental stages of common carp (Cyprinus carpio). The preparation PAX-18, with its active ingredient polyaluminium chloride (9% of Al), is a coagulation agent that is used mainly to precipitate phosphates, to prevent surface water eutrophication and incidences of cyanobacteria. It is applied to the water environment and thus could present a potential risk to fish. DESIGN: The toxicity tests were performed on common carp according to OECD (203, 210) methodologies. The acute toxic effect was evaluated for juveniles and the early development stage effect was observed in embryo-larval toxicity tests. The results of the toxicity tests (the number of dead individuals at particular test concentrations) were subjected to a probit analysis using an EKO-TOX 5.2 programme to determine the LC50 values of PAX-18. RESULTS: Acute toxicity value expressed as 96hLC50 was 753.1 +/- 24.3 mg.l-1 (67.8 mg.l-1 Al). Effect on early development stage expressed as the no observed effect concentration was 10 mg.l-1 (0.9 mg.l-1 Al), the lowest observed effect concentration was 50 mg.l-1 (4.5 mg.l 1 Al). No significant effects of the preparation PAX-18 in concentrations of 50 mg.l-1 of PAX-18 and lower were found on hatching, length and weight parameters, morphology and histopathology. CONCLUSION: The lethal concentration of PAX-18 found in acute toxicity tests on common carp was 7-14 times higher than the concentration which is usually applied to water (5-10 mg.l-1 Al). Moreover, fish in eutrophicated water sources are exposed to PAX-18 concentrations corresponding with the lowest observed effect concentration only for a short time, therefore the effect on them can be considered as minimal. PMID- 20027171 TI - Biochemical responses of juvenile and adult Japanese quails to cyanobacterial biomass. AB - OBJECTIVES: The aim of the present study was to evaluate differences between juvenile and adult Japanese quails in responses to the exposure to cyanobacterial biomass in the diet. DESIGN: The OECD 205 Guideline on Avian Dietary Toxicity Test (1984) was employed in the experiment. A total of 75 freshly hatched chicks and 30 adults were exposed to cyanobacterial biomass for 15 days and blood sampled daily and on days 5, 10 and 15, respectively. Japanese quail chicks and adults received the same daily dose of approximately 224.4 ng microcystins per gram of body weight. Biochemical responses were compared against controls. RESULTS: No Japanese quail chicks and adults died during the acute 15-day cyanobacterial-biomass exposure. Biochemical responses to the biomass in diet were first observed from day 5 post exposure to cyanobacterial biomass both in chicks and adults and there were age-related differences in the parameters changed. The responses of adult birds included an increase in lactate dehydrogenase, a drop in glucose and the total antioxidant capacity as well as a 15 to 20 % inhibition of acetylcholinesterase activity. Japanese quail chicks exposed to cyanobacterial biomass for the first 15 days after hatching reacted by having hypoproteinaemia, increased concentrations of triglycerides, uric acid and the total antioxidant capacity and a drop in high-density lipoprotein cholesterol in the blood. CONCLUSIONS: Chicks were not found to be more susceptible to the effects of biomass exposure. It seems that, due to their physiological preparation for the oxidative stress associated with hatching, Japanese quail chicks were even better able to cope with the cyanobacterial-biomass-induced oxidative stress than adults. PMID- 20027172 TI - Effects of cyanobacterial biomass on avian reproduction: a Japanese quail model. AB - OBJECTIVES: The present study was aimed at evaluation of the response of Japanese quails to cyanobacterial biomass administered in feed using biochemical profiles and parameters of reproduction. DESIGN: Effects of cyanobacterial biomass were studied according to the OECD 206 Guideline on Avian Reproduction Toxicity. A total of 16 control and 16 experimental pairs (32 males and 32 females) were analyzed. The chronic exposure of parent birds lasted eight weeks with the daily sum of 61.62 microg MCs including 26.54 microg MC-RR, 7.62 microg MC-YR and 27.39 microg MC-LR. RESULTS: There was no mortality both in control and cyanobacterial biomass-exposed adults during the present study. Nor did the birds show any clinical signs of intoxication. Lactate dehydrogenase activity was increased about three-fold in exposed birds. No other biochemical parameters were showing significant differences. A total of 824 and 821 eggs were laid by control and exposed birds, respectively, during the eight-week study period. Eggs laid by cyanobacterial-biomass-exposed hens had lower weight than in controls (11.99+/ 1.13g and 12.40+/-1.27g, respectively; p<0.01). Egg viability, hatchability, and the effect of hatching in control and experimental birds were 79.6+/-9.3 and 86.8+/-8.2% (p<0.05), 83.2+/-12.6 and 90.1+/-9.3%, and 65.2+/-17.7 and 77.7+/ 15.2% (p<0.05), respectively. There was also a statistically significant difference in the number of 14-day old survivors per hen per day in control and experimental birds (0.38+/-0.02 and 0.43+/-0.01 %, respectively). CONCLUSION: The lower weight of eggs produced by exposed parental hens was not reflected in their biological quality. On the contrary, reproductive parameters in cyanobacterial biomass-exposed birds were better than in the control group. It might be hypothesized that compounds of hormonal activity could be present in the complex cyanobacterial biomass. However, further research into this issue is necessary. PMID- 20027173 TI - Fish biochemical markers as a tool for pollution assessment on the Svitava and Svratka rivers, Czech Republic. AB - OBJECTIVES: The study was designed to assess the pollution of the Svitava and Svratka rivers in and around the industrial city of Brno (Czech Republic) by persistent organic pollutants using selected biochemical markers in chub. DESIGN: Levels of selected biochemical markers were measured in liver and plasma samples of chub. The concentrations of persistent organic pollutants (POPs) were determined in bottom sediment, semi-permeable membrane devices (SPMDs) and muscle samples, and consequently used for correlation with biochemical markers. RESULTS: Significant alterations (p < 0.05) in some biochemical markers were observed and associated with combined exposure to pollutants. The highest levels of pollutants were found at sites situated downstream from Brno. The most widespread changes were identified in the function of phase I detoxifying enzymes. Significant positive correlations were observed in cytochrome P450 content and DDT concentration in the semi-permeable membrane device (p = 0.019, rs = 0.886), and between ethoxyresorufin-O-deethylase activity and content of DDT (p = 0.041, rs = 0.352) and polychlorinated biphenyls (p = 0.034, rs = 0.365) in muscle tissues of indicator fish. CONCLUSION: The results presented in our study indicate the highest contamination of sites situated downstream from Brno, where the intensive industrial and agricultural activities as well as domestic waste and sewage most probably comprise the main impact sources of the enhanced level of pollutants and some biochemical markers in fish. PMID- 20027174 TI - Chronic health impairment due to 2,3,7,8-tetrachloro-dibenzo-p-dioxin exposure. AB - OBJECTIVES: The aim of this study, performed in 2008, was to evaluate the consequences of severe occupational intoxication with 2,3,7,8-tetrachloro-dibenzo p-dioxin (TCDD) that occurred during production of the herbicide trichlorophenoxyacetic acid in the period 1965-1968. DESIGN: Examination of 11 men, mean age 64.4+/-1.5 years, included: internal and neurological examination, eye fundus examination, TCDD in plasma, thyroid-stimulating hormone (TSH), testosterone and serum lipids, ultrasonography of the carotid artery, nerve conduction study (NCS), electroencephalography (EEG), visual evoked potential (VEP), Lanthony test of acquired visual impairment, single photon emission computer tomography (SPECT) of the brain, neuropsychological examination and carbohydrate-deficient transferrin (CDT), a marker of chronic ethanol intake. RESULTS: Mean TCDD level in 2008 was still 274.0+/-181.2 pg/g blood lipids (reference level is 2-3 pg/g). All (100%) patients had residues of chloracne/chloracne consequences, atherosclerotic changes on the eye fundus and plaques in the carotid arteries. Progression of intima-media thickness (IMT) from a mean of 0.84+/-0.14 mm in 2003 to 1.09+/-0.18 mm in 2008 was observed. Ninety one per cents of patients had impairment in SPECT of the brain; and 55% of patients had hyperfixation of the perfusion indicator as a measure of increased neuronal activity. Additionally, 91 % of patients were treated for hyperlipidaemia, 73 % for hypertension, 55 % for diabetes type 2, 45 % for ischemic heart disease, and 36 % for psychological disorders. The Lanthony test demonstrated acquired dyschromatopsia in 80 % of patients. Mean colour confusion index (CCI) was 1.438, which indicates impairment since 2003, when the index was 1.302. CDT was in the normal range and did not correlate with CCI. Neuropsychological status appeared stabilized in all 8 patients examined, with impairment in one or more parameter (memory, attention, verbal fluency, psychomotor speed, motorics) in comparison to the norm. CONCLUSION: Forty years after intoxication, the blood level of TCDD is still 100 times higher than in the general population. Other PCDD/Fs were not elevated. A high percentage of subjects suffer from neurological and vascular disorders. No association of alcohol consumption with neurological impairment was seen, and the highly significant correlation between CCI and TCDD blood concentration suggests that acquired colour impairment was associated with TCDD but not with alcohol consumption. IMT significantly increased during past 5 years. The patients obviously need complex treatment, including intense hypolipidaemic and antidepressant therapy. PMID- 20027175 TI - Induction of vitellogenin and gonadal impairment in chub (Leuciscus cephalus L.) after exposure to 17beta-estradiol and testosterone. AB - OBJECTIVES: A controlled laboratory study was carried out to quantify vitellogenin (VTG) concentrations in a common cyprinid freshwater fish, the chub (Leuciscus cephalus L.), exposed to steroid hormones. DESIGN: The effect of 17betaestradiol, testosterone and testosterone-estradiol mixture was investigated on vitellogenin induction. Gonad status was also determined. RESULTS: Oral exposure to estradiol and a testosterone-estradiol mixture increased (p < 0.01) blood plasma concentrations of VTG in blood plasma of both sexes. The testosterone-estradiol mixture had a negative effect on the investigated chub gonads. The effects were signified by histological changes when compared to control fish. CONCLUSION: Our results showed a significant VTG increase in blood plasma of both sexes, indicating that vitellogenic response in the chub is sensitive to steroid hormones. PMID- 20027176 TI - Effects of subchronic exposure to Successor 600 (pethoxamid 600 g L-1) on common carp Cyprinus carpio. AB - OBJECTIVES: We investigated the effects of the herbicide preparation Successor 600 on biochemical and haematological indices and the histolopathological parameters of common carp after 28 day exposure. Furthermore, the hepatosomatic index and induction of xenobiotic metabolizing enzymes and biomarkers were assessed. DESIGN: Juvenile fish were exposed to sublethal concentrations of Successor(R) 600 (0.06 mg L-1, 0.22 mg L-1 or 0.60 mg L-1) for 28 days. Haematological indices were assessed using unified methods of haematological examination in fish. Plasma biochemical indices were measured by biochemical analyzer, the vitellogenin concentration in male fish plasma was estimated by direct sandwich ELISA. In hepatopancreas, ethoxyresorufin-O-deethylase (EROD) activity was measured spectrofluorimetrically, concentration of total cytochrome P450 (CYP), glutathion (GSH) content and glutathion-S-transferase (GST) activity were determined spectrophotometrically. Histological changes in samples of hepatopancreas, skin, gills, spleen, head kidney and trunk kidney were examined by light microscopy. RESULTS: Haemoglobin, MCH and MCHC were significantly (p<0.05) reduced in fish treated with Successor(R) 600 of 0.22 and 0.60 mg L-1. LDH was enhanced (p<0.05), in the highest concentration of the preparation. Vitellogenin was detected in all male fish, with no difference between groups. HSI, GSH and GST were elevated (p<0.05), owing to the exposure, whereas CYP and EROD were not affected. Slight histopathological changes were demonstrated in skin, gills and hepatopancreas, with steroid tissue in head kidney samples of 0.60 mg L-1 treated fish. CONCLUSION: Successor 600 affected the haematological profile of the treated fish, while the effects on biochemical indices were less expressed. Male plasma vitellogenin concentrations were not indicative of estrogen disruptive effects after 28 days. The importance of GSH and GST for the metabolisation were demonstrated. In contrast, CYP and EROD were not influenced by any concentration tested. HSI was found to reflex pollution with Successor 600. Histopathological indices caused by the treatment were observed in various tissue samples of the treated fish. PMID- 20027177 TI - Effects of subchronic simazine exposure on some biometric, biochemical, hematological and histopathological parameters of common carp (Cyprinus carpio L.). AB - OBJECTIVES: This study assessed the subchronic effects of a triazine compound, simazine, on common carp (Cyprinus carpio L.) though, via by means of biometric, biochemical, hematological, and histological examination. DESIGN: One-year-old fish were exposed to simazine at four concentrations, 0.06, (reported concentration in Czech rivers), 4, 20, and 50 microg L-1 for 28 days and compared to carp in a non-treated control group. RESULTS: Exposure of fish to simazine at 0.06 microg L-1 had no effect on measured parameters. However, exposure to simazine at the concentrations of 4, 20, 50 microg L-1 showed significant differences in biochemical, hematological, and histopathological profiles of fish compared to controls. CONCLUSION: Our data suggest that simazine in the recorded environmental concentration 0.06 microg L-1 had no effect on common carp. Subchronic exposure to 4, 20, and 50 microg L-1 of simazine was associated with alterations in biochemical and hematological indices and in fish organ tissues. PMID- 20027178 TI - Terbutryn toxicity to Danio rerio: effects of subchronic exposure on fish growth. AB - OBJECTIVES: The aim of this study was to investigate effects of subchronic exposure to sublethal levels of terbutryn on growth and on histopathological changes in Danio rerio. DESIGN: The acute toxicity tests were performed on the juvenile stage of Danio rerio according to OECD No. 203 Fish, Acute Toxicity Test. The juvenile growth tests were performed on D. rerio according to the OECD method No. 215. Fish at the age of 20 days were exposed to the terbutryn environmental concentration commonly detected in the Czech rivers (0.02 microg L 1) and the range of sublethal concentrations of terbutryn (0.06, 0.2, 0.6 and 2 mg L-1) for 28 days. RESULTS: The 96hLC50 terbutryn mean value for the juvenile stage of D. rerio was 5.71 +/- 0.46 mg L-1. A significant decrease (p < 0.01) in the fish growth caused by terbutryn was observed in the concentration of 0.6 mg L 1. The value of NOEC was 0.2 mg L-1 and LOEC was 0.6 mg L-1 of terbutryn. We found the damage to tubular system of kidneys in the concentration of 0.6 mg L-1 of terbutryn. CONCLUSION: The environmental concentration of terbutryn in the Czech rivers did not have any effects on growth and on histopathological changes in D. rerio and this concentration is lower than determined LOEC and NOEC values of terbutryn. PMID- 20027179 TI - Comparison of the sensitivity of different fish species to medical substances. AB - OBJECTIVES: Formaldehyde, sodium chloride and potassium permanganate belong to the commonly used substances for fish treatment. The aim is to define and compare their acute toxicity and therapeutic index between two fish species - Danio rerio and Poecilia reticulata. DESIGN: To determine acute toxicity of these compounds, the semistatic method was implemented in compliance with the OECD No. 203 (Fish acute toxicity test). In each test series, 4 acute toxicity tests were performed. The results were subjected to the probit analysis to determine the 96hLC50 values. Therapeutic index (TI) was calculated with respect to short-term LC50 and effective therapeutic concentrations commonly used for fish treatment. RESULTS: The mean 96hLC50 values for sodium chloride were 21.69 +/- 0.92 g l-1 (TI = 1.4) for P. reticulata and 10.39 +/- 0.12 g l-1 (TI = 1.0) for D. rerio. The acute toxicity of formaldehyde for D. rerio expressed as 96LC50 was 0.12 +/- 0.003 ml l 1 (TI = 5.24) and for P. reticulata 0.1 +/- 0.003 ml l-1 (TI = 4.9). The acute toxicity of potassium permanganate for D. rerio expressed as 96LC50 was 1.25 +/- 0.15 mg l-1(TI = 1.5) and for P. reticulata 1.43 +/- 0.05 mg l-1 (TI = 2). CONCLUSION: P. reticulata showed significantly higher (p < 0.05) tolerance to sodium chloride than D. rerio, whereas D. rerio showed significantly higher (p < 0.05) tolerance to formaldehyde than P. reticulata. The acute toxicity of potassium permanganate was comparable for both fish species. Calculated therapeutic indexes of all tested substances were low; therefore it is important to conduct preliminary tolerance tests before application of treatment baths. PMID- 20027180 TI - Preparation of embryos for electron microscopy of the Drosophila embryonic heart tube. AB - The morphogenesis of the Drosophila embryonic heart tube has emerged as a valuable model system for studying cell migration, cell-cell adhesion and cell shape changes during embryonic development. One of the challenges faced in studying this structure is that the lumen of the heart tube, as well as the membrane features that are crucial to heart tube formation, are difficult to visualize in whole mount embryos, due to the small size of the heart tube and intra-lumenal space relative to the embryo. The use of transmission electron microscopy allows for higher magnification of these structures and gives the advantage of examining the embryos in cross section, which easily reveals the size and shape of the lumen. In this video, we detail the process for reliable fixation, embedding, and sectioning of late stage Drosophila embryos in order to visualize the heart tube lumen as well as important cellular structures including cell-cell junctions and the basement membrane. PMID- 20027181 TI - The role of hypoxia and neurogenic genes (Mash-1 and Prox-1) in the developmental programming and maturation of pulmonary neuroendocrine cells in fetal mouse lung. AB - Pulmonary neuroendocrine cells (PNECs) are the first cell type to differentiate within the primitive airway epithelium, suggesting a possible role in lung development. The differentiation of PNECs in fetal lung is governed by proneural genes such as the mammalian homolog of the achaete-scute complex (Mash-1) and a related transcription factor, hairy and enhancer of split1 (Hes-1). We examined the expression of Mash-1 and a downstream transcription factor Prox-1 in the developing mouse lung of wild-type and respective knockout mouse models. During early stages (embryonic day 12, E12) of development, only some PNECs expressed Mash-1 and Prox-1, but by E15, all PNECs coexpressed both transcription factors. PNECs failed to develop in Mash-1 but not in Prox-1-null mice, indicating that Mash-1 is essential for the initiation of the PNEC phenotype, whereas Prox-1 is associated with the development of this phenotype. As lung develops within a low O(2) environment (fetal euoxia, pO(2) approximately 20 to 30 mm Hg), we examined the effects of hypoxia on PNEC differentiation. Organ cultures of fetal mouse lungs at E12 and E16 were maintained under either 20% O(2) (normoxia, Nox) or 5% O(2) (hypoxia, Hox) and were examined every 24 h for up to 6 days in culture. In E12 explants, Hox enhanced branching morphogenesis and increased cell proliferation, but PNEC numbers and Mash-1 expression were significantly reduced. This effect could be reversed by switching the explants back to Nox. In contrast, Hox had no apparent effect on Hes-1 expression. Similarly, Hox had no effect on airway branching, PNEC numbers, or Mash-1 expression in E16 explants, indicating locked-in developmental programming. We suggest that during early stages of lung development, pO(2) concentration in concert with neurogenic gene expression modulates PNEC phenotype. Thus, disturbances in intrauterine pO(2) homeostasis could alter the functional maturation of the PNEC system and hence be involved in the pathogenesis of various perinatal pulmonary disorders. PMID- 20027182 TI - Reprogramming towards pluripotency requires AID-dependent DNA demethylation. AB - Reprogramming of somatic cell nuclei to yield induced pluripotent stem (iPS) cells makes possible derivation of patient-specific stem cells for regenerative medicine. However, iPS cell generation is asynchronous and slow (2-3 weeks), the frequency is low (<0.1%), and DNA demethylation constitutes a bottleneck. To determine regulatory mechanisms involved in reprogramming, we generated interspecies heterokaryons (fused mouse embryonic stem (ES) cells and human fibroblasts) that induce reprogramming synchronously, frequently and fast. Here we show that reprogramming towards pluripotency in single heterokaryons is initiated without cell division or DNA replication, rapidly (1 day) and efficiently (70%). Short interfering RNA (siRNA)-mediated knockdown showed that activation-induced cytidine deaminase (AID, also known as AICDA) is required for promoter demethylation and induction of OCT4 (also known as POU5F1) and NANOG gene expression. AID protein bound silent methylated OCT4 and NANOG promoters in fibroblasts, but not active demethylated promoters in ES cells. These data provide new evidence that mammalian AID is required for active DNA demethylation and initiation of nuclear reprogramming towards pluripotency in human somatic cells. PMID- 20027183 TI - Human host factors required for influenza virus replication. AB - Influenza A virus is an RNA virus that encodes up to 11 proteins and this small coding capacity demands that the virus use the host cellular machinery for many aspects of its life cycle. Knowledge of these host cell requirements not only informs us of the molecular pathways exploited by the virus but also provides further targets that could be pursued for antiviral drug development. Here we use an integrative systems approach, based on genome-wide RNA interference screening, to identify 295 cellular cofactors required for early-stage influenza virus replication. Within this group, those involved in kinase-regulated signalling, ubiquitination and phosphatase activity are the most highly enriched, and 181 factors assemble into a highly significant host-pathogen interaction network. Moreover, 219 of the 295 factors were confirmed to be required for efficient wild type influenza virus growth, and further analysis of a subset of genes showed 23 factors necessary for viral entry, including members of the vacuolar ATPase (vATPase) and COPI-protein families, fibroblast growth factor receptor (FGFR) proteins, and glycogen synthase kinase 3 (GSK3)-beta. Furthermore, 10 proteins were confirmed to be involved in post-entry steps of influenza virus replication. These include nuclear import components, proteases, and the calcium/calmodulin dependent protein kinase (CaM kinase) IIbeta (CAMK2B). Notably, growth of swine origin H1N1 influenza virus is also dependent on the identified host factors, and we show that small molecule inhibitors of several factors, including vATPase and CAMK2B, antagonize influenza virus replication. PMID- 20027184 TI - The nuts and bolts of AGC protein kinases. AB - The AGC kinase subfamily of protein kinases contains 60 members, including PKA, PKG and PKC. The family comprises some intensely examined protein kinases (such as Akt, S6K, RSK, MSK, PDK1 and GRK) as well as many less well-studied enzymes (such as SGK, NDR, LATS, CRIK, SGK494, PRKX, PRKY and MAST). Research has shed new light onto the architecture and regulatory mechanisms of these kinases. In addition, AGC kinases mediate diverse and important cellular functions, and their mutation and/or dysregulation contributes to the pathogenesis of many human diseases, including cancer and diabetes. PMID- 20027185 TI - Regulation of cell signalling by uPAR. AB - Urokinase-type plasminogen activator receptor (uPAR) expression is elevated during inflammation and tissue remodelling and in many human cancers, in which it frequently indicates poor prognosis. uPAR regulates proteolysis by binding the extracellular protease urokinase-type plasminogen activator (uPA; also known as urokinase) and also activates many intracellular signalling pathways. Coordination of extracellular matrix (ECM) proteolysis and cell signalling by uPAR underlies its important function in cell migration, proliferation and survival and makes it an attractive therapeutic target in cancer and inflammatory diseases. uPAR lacks transmembrane and intracellular domains and so requires transmembrane co-receptors for signalling. Integrins are essential uPAR signalling co-receptors and a second uPAR ligand, the ECM protein vitronectin, is also crucial for this process. PMID- 20027186 TI - Mechanisms guiding primordial germ cell migration: strategies from different organisms. AB - The regulated migration of cells is essential for development and tissue homeostasis, and aberrant cell migration can lead to an impaired immune response and the progression of cancer. Primordial germ cells (PGCs), precursors to sperm and eggs, have to migrate across the embryo to reach somatic gonadal precursors, where they carry out their function. Studies of model organisms have revealed that, despite important differences, several features of PGC migration are conserved. PGCs require an intrinsic motility programme and external guidance cues to survive and successfully migrate. Proper guidance involves both attractive and repulsive cues and is mediated by protein and lipid signalling. PMID- 20027187 TI - Emerging role for the cytoskeleton as an organizer and regulator of translation. AB - The cytoskeleton is an intricate and dynamic fibrous network that has an essential role in the generation and regulation of cell architecture and cellular mechanical properties. The cytoskeleton also evolved as a scaffold that supports diverse biochemical pathways. Recent evidence favours the hypothesis that the cytoskeleton participates in the spatial organization and regulation of translation, at both the global and local level, in a manner that is crucial for cellular growth, proliferation and function. PMID- 20027188 TI - Imaging approaches for the study of cell-based cardiac therapies. AB - Despite promising preclinical data, the treatment of cardiovascular diseases using embryonic, bone-marrow-derived, and skeletal myoblast stem cells has not yet come to fruition within mainstream clinical practice. Major obstacles in cardiac stem cell investigations include the ability to monitor cell engraftment and survival following implantation within the myocardium. Several cellular imaging modalities, including reporter gene and MRI-based tracking approaches, have emerged that provide the means to identify, localize, and monitor stem cells longitudinally in vivo following implantation. This Review will examine the various cardiac cellular tracking modalities, including the combinatorial use of several probes in multimodality imaging, with a focus on data from the past 5 years. PMID- 20027189 TI - The conundrum of detecting stable angina pectoris in the community setting. AB - Individuals with undetected stable angina pectoris (SAP) as a consequence of undiagnosed coronary artery disease are at high risk of poor quality of life and a premature fatal event (for example, sudden cardiac death out of hospital). If the extent and distribution of SAP are accurately identified at the population level, clinical screening could potentially be targeted and evaluated to optimize the management and secondary prevention of underlying coronary artery disease. Common measures of SAP in populations have important limitations. Measures chosen to identify such cases should reflect their validity as measures of undiagnosed SAP, currently symptomatic angina or lifetime diagnosis of angina. PMID- 20027190 TI - The role of SRC-3 in human breast cancer. AB - Members of the nuclear receptor superfamily are ligand-regulated transcription factors involved in the control of a broad range of normal physiological and disease processes. The estrogen receptor alpha (ERalpha) is a member of the steriod receptor family, which is part of the nuclear receptor superfamily. ERalpha it is important for many biological processes and plays a key role in the pathogenesis of breast cancer. Gene regulation by ERalpha requires the recruitment of a multitude of transcriptional co-regulators to the promoters of estrogen-responsive genes. There is evidence in support of the involvement of these co-regulators in breast cancer progression. We review the role of steroid receptor co-activator-3 (SRC-3), which is frequently amplified in breast cancer, and its role in breast cancer risk, outcome and response to endocrine therapy in patients with breast cancer. PMID- 20027191 TI - Molecular predictors of response to trastuzumab and lapatinib in breast cancer. AB - Trastuzumab is a monoclonal antibody directed against the human EGFR2 (HER2) protein that has been shown to improve survival in patients with HER2-positive breast cancer. Lapatinib is an oral small-molecule tyrosine kinase inhibitor directed against EGFR and HER2. Lapatinib therapy was shown to prolong the time to progression and increase the rate of response to capecitabine in patients who had received anthracycline-based and taxane-based chemotherapy, and whose tumors had progressed on trastuzumab. HER2 status, either gene copy number or the protein expression level, is the best predictive marker available for assessing response to trastuzumab and lapatinib. Whether the power of this predictive marker is the same in advanced and early-stage cancers is unknown. There is great interest in developing diagnostic tests that predict which patients are more likely to benefit from specific HER2-directed therapies. Novel therapeutics that will overcome resistance to trastuzumab and lapatinib are under intense clinical development. In the future, it will be important to characterize mechanisms of resistance in metastatic tumors to determine which novel targeted therapy will be most appropriate for individual patients. PMID- 20027192 TI - Fluid balance and acute kidney injury. AB - Intravenous fluids are widely administered to patients who have, or are at risk of, acute kidney injury (AKI). However, deleterious consequences of overzealous fluid therapy are increasingly being recognized. Salt and water overload can predispose to organ dysfunction, impaired wound healing and nosocomial infection, particularly in patients with AKI, in whom fluid challenges are frequent and excretion is impaired. In this Review article, we discuss how interstitial edema can further delay renal recovery and why conservative fluid strategies are now being advocated. Applying these strategies in critical illness is challenging. Although volume resuscitation is needed to restore cardiac output, it often leads to tissue edema, thereby contributing to ongoing organ dysfunction. Conservative strategies of fluid management mandate a switch towards neutral balance and then negative balance once hemodynamic stabilization is achieved. In patients with AKI, this strategy might require renal replacement therapy to be given earlier than when more-liberal fluid management is used. However, hypovolemia and renal hypoperfusion can occur in patients with AKI if excessive fluid removal is pursued with diuretics or extracorporeal therapy. Thus, accurate assessment of fluid status and careful definition of targets are needed at all stages to improve clinical outcomes. A conservative strategy of fluid management was recently tested and found to be effective in a large, randomized, controlled trial in patients with acute lung injury. Similar randomized, controlled studies in patients with AKI now seem justified. PMID- 20027194 TI - Advising the government. The dismissal of a senior science advisor in the UK has lead to a debate on the role of expert advice in crafting public policy. PMID- 20027195 TI - Building consensus. A group of Golgi researchers tackle controversies in the field head-on and emerge with a blueprint for future research. PMID- 20027196 TI - Accidental encounters: the chance to solve a mystery. PMID- 20027193 TI - Aldosterone and arterial hypertension. AB - In the setting of primary aldosteronism, elevated aldosterone levels are associated with increased blood pressure. Aldosterone concentrations within the normal range, however, can also alter blood pressure. Furthermore, the aldosterone-to-renin ratio, an indicator of aldosterone excess, is associated with hypertension, even in patients without excessive absolute aldosterone levels. In this Review we assess the data on the role of aldosterone in the development and maintenance of hypertension. We provide an overview of the complex crosstalk between genetic and environmental factors, and about aldosterone-mediated arterial hypertension and target organ damage. The discussion is organized according to major targets of aldosterone action: the collecting duct in the kidney, the vasculature and the central nervous system. The antihypertensive efficacy of mineralocorticoid-receptor blockers, even in patients with aldosterone values in the normal range, supports the evidence that aldosterone plays a part in blood pressure elevation in the absence of primary aldosteronism. PMID- 20027197 TI - Two Rabs for exosome release. AB - Exosomes are endosome-derived membrane vesicles that are key for intercellular communication in the immune system and elsewhere. Rab27A and Rab27B GTPases and two of their cognate effector proteins seem to be needed to drive the physiologically important exosome-release process in certain cell types. PMID- 20027198 TI - Tensions divide. AB - During development, proliferating cells are organized into compartments with boundaries across which cells fail to intermix. Compartment boundaries are often attributed to differential cell-cell adhesion between separate compartments. However, tension generated by actomyosin cables at boundaries can also function as a barrier that prevents cell mixing. PMID- 20027199 TI - Myc and a Cdk2 senescence switch. AB - Cdk2 has been shown to have an unanticipated role in suppressing Myc-induced senescence. This has implications for how c-Myc overcomes failsafe mechanisms to induce tumorigenesis and suggests that the inhibition of Cdk2 may have therapeutic efficacy in the treatment of cancer. PMID- 20027201 TI - A new FFT-based algorithm to compute Born radii in the generalized Born theory of biomolecule solvation. AB - In this paper, a new method for calculating effective atomic radii within the generalized Born (GB) model of implicit solvation is proposed, for use in computer simulations of bio-molecules. First, a new formulation for the GB radii is developed, in which smooth kernels are used to eliminate the divergence in volume integrals intrinsic in the model. Next, the Fast Fourier Transform (FFT) algorithm is applied to integrate smoothed functions, taking advantage of the rapid spectral decay provided by the smoothing. The total cost of the proposed algorithm scales as O(N(3)logN + M) where M is the number of atoms comprised in a molecule, and N is the number of FFT grid points in one dimension, which depends only on the geometry of the molecule and the spectral decay of the smooth kernel but not on M. To validate our algorithm, numerical tests are performed for three solute models: one spherical object for which exact solutions exist and two protein molecules of differing size. The tests show that our algorithm is able to reach the accuracy of other existing GB implementations, while offering much lower computational cost. PMID- 20027202 TI - A Fast Hermite Transform. AB - We present algorithms for fast and stable approximation of the Hermite transform of a compactly supported function on the real line, attainable via an application of a fast algebraic algorithm for computing sums associated with a three-term relation. Trade-offs between approximation in bandlimit (in the Hermite sense) and size of the support region are addressed. Numerical experiments are presented that show the feasibility and utility of our approach. Generalizations to any family of orthogonal polynomials are outlined. Applications to various problems in tomographic reconstruction, including the determination of protein structure, are discussed. PMID- 20027203 TI - Synthetic studies of neoclerodane diterpenoids from Salvia splendens and evaluation of Opioid Receptor affinity. AB - Salvinorin A (1), a neoclerodane diterpene from the hallucinogenic mint Salvia divinorum, is the only known non-nitrogenous and specific kappa-opioid agonist. Several structural congeners of 1 isolated from Salvia splendens (2 - 8) together with a series of semisynthetic derivatives (9 - 24), some of which possess a pyrazoline structural moiety (9, 19 - 22), have been tested for affinity at human mu, delta, and kappa opioid receptors. None of these compounds showed high affinity binding to these receptors. However, 10 showed modest affinity for kappa receptors suggesting other naturally neoclerodanes from different Salvia species may possess opioid affinity. PMID- 20027204 TI - Impact of methicillin-resistant Staphylococcus aureus on treatment of hand infections. AB - OBJECTIVE: With the increasing incidence of methicillin-resistant Staphylococcus aureus (MRSA) in hand infections in urban centers, multiple studies have recommended using MRSA-effective antibiotics as first-line treatment of hand infections. This study assesses the effect of adopting this recommendation for the treatment of hand infections at the authors' hospital. METHODS: Patients with hand infections drained in the authors' hospital were prospectively enrolled in an observational study over a 12-month period. Culture results and response to treatment were recorded. RESULTS: Twenty-two patients met inclusion criteria. Eleven of 14 patients with S aureus infections had MRSA. All of these patients responded to the initial antibiotic selected. Two patients had infections that did not respond to trimethoprim-sulfamethoxazole. One grew group A Streptococcus infection, and the other had lymphangitic streaking that suggested Streptococcus infection. CONCLUSION: Because of the high prevalence of MRSA among hand infections at the authors' institution, we continue to prescribe MRSA-effective antibiotics as first-line treatment of hand abscesses. Close follow-up is still necessary to confirm that each patient has responded appropriately to treatment or to allow modification of the treatment plan if the patient has not responded to treatment. PMID- 20027205 TI - Distributed fading memory for stimulus properties in the primary visual cortex. AB - It is currently not known how distributed neuronal responses in early visual areas carry stimulus-related information. We made multielectrode recordings from cat primary visual cortex and applied methods from machine learning in order to analyze the temporal evolution of stimulus-related information in the spiking activity of large ensembles of around 100 neurons. We used sequences of up to three different visual stimuli (letters of the alphabet) presented for 100 ms and with intervals of 100 ms or larger. Most of the information about visual stimuli extractable by sophisticated methods of machine learning, i.e., support vector machines with nonlinear kernel functions, was also extractable by simple linear classification such as can be achieved by individual neurons. New stimuli did not erase information about previous stimuli. The responses to the most recent stimulus contained about equal amounts of information about both this and the preceding stimulus. This information was encoded both in the discharge rates (response amplitudes) of the ensemble of neurons and, when using short time constants for integration (e.g., 20 ms), in the precise timing of individual spikes (/= 140 mmHg or diastolic BP >/= 90 mmHg or on treatment with BP medications). A more conservative definition of high BP (systolic BP >/= 160 mmHg or diastolic BP >/= 100 mmHg) substantially reduced the prevalence rate. The marked differences in the proportion of the populations on high BP medication (range between 0.6 and 10.8%) raised problems in comparing the prevalence of high BP across sites when using the commonly used definition of high BP as in this study. In the four HDSS in Bangladesh, women had a higher prevalence of high BP than men; the reverse was true in the other sites (Chililab, Filabavi in Vietnam; Kanchanaburi, Thailand; and Vadu, India) where men experienced higher prevalence than women. Overweight and obesity were significantly associated with high BP, with odds ratio ranging from two in Chililab to five in Filabavi (both in Vietnam HDSS). CONCLUSION: The patterns of BP in these nine cross-sectional surveys were complex, reflecting the fact that the Asian countries are at different stages of the epidemiological transition. Actions to prevent the rise of BP levels are urgently required. An emphasis should be placed on cost-effective interventions to reduce salt consumption in the population as an immediate priority. PMID- 20027256 TI - Self-reported use of tobacco products in nine rural INDEPTH Health and Demographic Surveillance Systems in Asia. AB - BACKGROUND: Tobacco use is the most preventable cause of premature death and disability. Even though tobacco use is common in many Asian countries, reliable and comparable data on the burden imposed by tobacco use in this region are sparse, and surveillance systems to track trends are in their infancy. OBJECTIVE: To assess and compare the prevalence of tobacco use and its associated factors in nine selected rural sites in five Asian countries. METHODS: Tobacco use among 9,208 men and 9,221 women aged 25-64 years in nine Health and Demographic Surveillance System (HDSS) sites in five Asian countries of the INDEPTH Network were examined in 2005 as part of a broader survey of the major chronic non communicable disease risk factors. All sites used a standardised protocol based on the WHO STEPS approach to risk factor surveillance; expanded questions of local relevance, including chewing tobacco, were also included. Multivariable logistic regression was used to assess demographic factors associated with tobacco use. RESULTS: Tobacco use, whether smoked or chewed, was common across all sites with some notable variations. More than 50% of men smoked daily; this applied to almost all age groups. Few women smoked daily in any of the sites. However, women were more likely to chew tobacco than men in all sites except Vadu in India. Tobacco use in men began in late adolescence in most of the sites and the number of cigarettes smoked daily ranged from three to 15. Use of both forms of tobacco, smoked and chewed, was associated with age, gender and education. Men were more likely to smoke compared to women, smoking increased with age in the four sites in Bangladesh but not in other sites and with low level of education in all the sites. CONCLUSION: The prevalence of tobacco use, regardless of the type of tobacco, was high among men in all of these rural populations with tobacco use started during adolescence in all HDSS sites. Innovative communication strategies for behaviour change targeting adolescents in schools and adult men and women at work or at home, may create a mass awareness about adverse health consequences of tobacco smoking or chewing tobacco. Such efforts, to be effective, however, need to be supported by strong legislation and leadership. Only four of the five countries involved in this multi-site study have ratified the Framework Convention on Tobacco Control, and even where it has been ratified, implementation is uneven. PMID- 20027257 TI - Social factors and overweight: evidence from nine Asian INDEPTH Network sites. AB - BACKGROUND: Overweight/obesity increases the risk of morbidity and mortality from a number of chronic conditions, including heart disease, stroke, diabetes and some cancers. This study examined the distribution of body mass index (BMI) in nine Health and Demographic Surveillance System (HDSS) sites in five Asian countries and investigated the association between social factors and overweight. DATA AND METHODS: This cross-sectional study was conducted in nine HDSS sites in Bangladesh, India, Indonesia, Thailand and Vietnam. The methodology of the WHO STEPwise approach to Surveillance with core risk factors (Step 1) and physical measurements for weight, height and waist circumference (Step 2) were included. In each site, about 2,000 men and women aged 25-64 years were selected randomly using the HDSS database. Weight was measured using electronic scales, height was measured by portable stadiometers and waist circumference was measured by measuring tape. Overweight/obesity was assessed by BMI defined as the weight in kilograms divided by the square of the height in metres (kg/m(2)). RESULTS: At least 10% people were overweight (BMI >/= 25) in each site except for the two sites in Vietnam and WATCH HDSS in Bangladesh where few men and women were overweight. After controlling for all the variables in the model, overweight increases with age initially and then declines, with increasing education, and with gender with women being heavier than men. People who eat vegetables and fruits below the recommended level and those who do high level of physical activity are, on the whole, less heavy than those who eat more and do less physical activity. CONCLUSIONS: As the proportion of the population classified as being overweight is likely to increase in most sites and overweight varies by age, sex, and social and behavioural factors, behavioural interventions (physical exercise, healthy diet) should be developed for the whole population together with attention to policy around nutrition and the environment, in order to reduce the adverse effects of overweight on health. PMID- 20027258 TI - A role for INDEPTH Asian sites in translating research to action for non communicable disease prevention and control: a case study from Ballabgarh, India. AB - BACKGROUND: The International Network of field sites with continuous Demographic Evaluation of Populations and Their Health (INDEPTH) has 34 Health and Demographic Surveillance System (HDSS) in 17 different low and middle-income countries. Of these, 23 sites are in Africa, 10 sites are in Asia, and one in Oceania. The INDEPTH HDSS sites in Asia identified chronic non-communicable diseases (NCDs) as a neglected area of attention. As a first step, they conducted NCD risk factor surveys within nine sites in five countries. These sites are now looking to broaden the agenda of research on NCDs using the baseline data to inform policy and practice. METHODS: A conceptual framework for translating research into action for NCDs at INDEPTH sites was developed. This had five steps - assess the problem, understand the nature of the problem, evaluate different interventions in research mode, implement evidence-based interventions in programme mode, and finally, share knowledge and provide leadership to communities and countries. Ballabgarh HDSS site in India has successfully adopted these steps and is used as a case study to demonstrate how this progress was achieved and what factors were responsible for a successful outcome. RESULTS: Most of the HDSS sites are in the second step of the process of translating research to action (understand the problem). The conduct of NCD risk factor surveys has enabled an assessment of the burden of NCD risk together with determinants in order to understand the burden at the population level. The experience from Ballabgarh HDSS exemplifies that the following steps - pilot testing the interventions, implementing activities in programme mode, and finally, share knowledge and provide leadership - are also possible in rural settings in low-income countries. The critical success factors identified were involvement of a premier medical institution, pre-existing links to policy makers and programme managers, strong commitment of the HDSS team and adequate human resource capacity. CONCLUSION: All INDEPTH HDSS sites now need to strengthen their links to health systems at different levels and enhance their capacity to engage different stakeholders in their respective country settings so as to translate the current knowledge into actions that can benefit the health of the population they serve and beyond. PMID- 20027259 TI - Fruit and vegetable consumption in rural adults population in INDEPTH HDSS sites in Asia. AB - BACKGROUND: Low fruit and vegetable consumption is among the top 10 risk factors contributing to mortality worldwide. WHO/FAO recommends intake of a minimum of 400 grams (or five servings) of fruits and vegetables per day for the prevention of chronic diseases such as heart diseases, cancer, diabetes, and obesity. OBJECTIVE: This paper examines the fruit and vegetable consumption patterns and the prevalence of inadequate fruit and vegetable consumption (less than five servings a day) among the adult population in rural surveillance sites in five Asian countries. DATA AND METHODS: The analysis is based on data from a 2005 cross-site study on non-communicable disease risk factors which was conducted in nine Asian INDEPTH Health and Demographic Surveillance System (HDSS) sites. Standardised protocols and methods following the WHO STEPwise approach to risk factor surveillance were used. The total sample was 18,429 adults aged 25-64 years. Multivariate logistic regression analysis was performed to assess the association between socio-demographic factors and inadequate fruit and vegetable consumption. RESULTS: Inadequate fruit and vegetable consumption was common in all study sites. The proportions of inadequate fruit and vegetable consumption ranged from 63.5% in men and 57.5% in women in Chililab HDSS in Vietnam to the whole population in Vadu HDSS in India, and WATCH HDSS in Bangladesh. Multivariate logistic regression analysis in six sites, excluding WATCH and Vadu HDSS, showed that being in oldest age group and having low education were significantly related to inadequate fruit and vegetable consumption, although the pattern was not consistent through all six HDSS. CONCLUSIONS: Since such a large proportion of adults in Asia consume an inadequate amount of fruits and vegetables, despite of the abundant availability, education and behaviour change programmes are needed to promote fruit and vegetable consumption. Accurate and useful information about the health benefits of abundant fruit and vegetable consumption should be widely disseminated. PMID- 20027260 TI - Clustering of chronic non-communicable disease risk factors among selected Asian populations: levels and determinants. AB - BACKGROUND: The major chronic non-communicable diseases (NCDs) operate through a cluster of common risk factors, whose presence or absence determines not only the occurrence and severity of the disease, but also informs treatment approaches. Primary prevention based on mitigation of these common risk factors through population-based programmes is the most cost-effective approach to contain the emerging epidemic of chronic NCDs. OBJECTIVES: This study was conducted to explore the extent of risk factors clustering for the major chronic NCDs and its determinants in nine INDEPTH Health and Demographic Surveillance System (HDSS) sites of five Asian countries. DESIGN: Data originated from a multi-site chronic NCD risk factor prevalence survey conducted in 2005. This cross-sectional survey used a standardised questionnaire developed by the WHO to collect core data on common risk factors such as tobacco use, intake of fruits and vegetables, physical inactivity, blood pressure levels, and body mass index. Respondents included randomly selected sample of adults (25-64 years) living in nine rural HDSS sites in Bangladesh, India, Indonesia, Thailand, and Vietnam. RESULTS: Findings revealed a substantial proportion (>70%) of these largely rural populations having three or more risk factors for chronic NCDs. Chronic NCD risk factors clustering was associated with increasing age, being male, and higher educational achievements. Differences were noted among the different sites, both between and within country. CONCLUSIONS: Since there is an extensive clustering of risk factors for the chronic NCDs in the populations studied, the interventions also need to be based on a comprehensive approach rather than on a single factor to forestall its cumulative effects which occur over time. This can work best if it is integrated within the primary health care system and the HDSS can be an invaluable epidemiological resource in this endeavor. PMID- 20027261 TI - Prevalence of physical inactivity in nine rural INDEPTH Health and Demographic Surveillance Systems in five Asian countries. AB - BACKGROUND: Physical inactivity leads to higher morbidity and mortality from chronic non-communicable diseases (NCDs) such as stroke and heart disease. In high income countries, studies have measured the population level of physical activity, but comparable data are lacking from most low and middle-income countries. OBJECTIVE: To assess the level of physical inactivity and its associated factors in selected rural sites in five Asian countries. METHODS: The multi-site cross-sectional study was conducted in nine rural Health and Demographic Surveillance System (HDSS) sites within the INDEPTH Network in Bangladesh, India, Indonesia, Thailand, and Vietnam. Using the methodology from the WHO STEPwise approach to Surveillance (STEPS), about 2,000 men and women aged 25-64 years were selected randomly from each HDSS sampling frame. Physical activity at work and during leisure time, and on travel to and from places, was measured using the Global Physical Activity Questionnaire version 2 (GPAQ2). The total activity was calculated as the sum of the time spent in each domain of activities in metabolic equivalent-minutes per week, and was used to determine the level of physical activity. Multivariable logistic regression was used to assess demographic factors associated with a low level of physical activity. RESULTS: The prevalence of physical inactivity ranged from 13% in Chililab HDSS in Vietnam to 58% in Filabavi HDSS in Vietnam. The majority of men were physically active, except in the two sites in Vietnam. Most of the respondents walked or cycled for at least 10 minutes to get from place to place, with some exceptions in the HDSSs in Indonesia and Thailand. The majority of respondents, both men and women, were inactive during their leisure time. Women, older age, and high level of education were significantly associated with physical inactivity. CONCLUSION: This study showed that over 1/4 men and 1/3 women in Asian HDSSs within the INDEPTH Network are physically inactive. The wide fluctuations between the two HDSS in Vietnam offer an opportunity to explore further urbanisation and environmental impacts on physical activity. Considering the importance of physical activity in improving health and preventing chronic NCDs, efforts need to be made to promote physical activity particularly among women, older people, and high education groups in these settings. PMID- 20027263 TI - Ethnomedicine in healthcare systems of the world: a Semester at Sea pilot survey in 11 countries. AB - BACKGROUND: An understanding and appreciation for the varied healthcare systems in use throughout the world are increasingly vital for medical personnel as patient populations are now composed of ethnically diverse people with wide ranging belief systems. OBJECTIVE: While not a statistically valid survey, this pilot study gives a global overview of healthcare differences around the world. DESIGN: A pilot study of 459 individuals from 11 different countries around the world was administered by 33 students in the upper division course, People, Pathology, and World Medicine from Semester at Sea, Fall 2007, to ascertain trends in healthcare therapies. Open-ended surveys were conducted in English, through an interpreter, or in the native language. RESULTS: Western hospital use ranked highly for all countries, while ethnomedical therapies were utilized to a lesser degree. Among the findings, mainland China exhibited the greatest overall percentage of ethnomedical therapies, while the island of Hong Kong, the largest use of Western hospitals. CONCLUSIONS: The figures and trends from the surveys suggest the importance of understanding diverse cultural healthcare beliefs when treating individuals of different ethnic backgrounds. The study also revealed the increasingly complex and multisystem-based medical treatments being used internationally. PMID- 20027262 TI - Using the INDEPTH HDSS to build capacity for chronic non-communicable disease risk factor surveillance in low and middle-income countries. AB - BACKGROUND: Chronic non-communicable diseases (NCDs) are the leading cause of morbidity, mortality, and disability worldwide. More than 80% of chronic disease deaths occur in low-income and middle-income countries. Epidemiological data on the burden of chronic NCD and the risk factors which predict them are lacking in most low-income countries. The INDEPTH Network (http://www.indepth-network.org) which includes the Health and Demographic Surveillance System (HDSS) with many surveillance sites in low-middle income countries provided an opportunity to establish surveillance of the major chronic NCD risk factors in 2005 using a standardised approach. OBJECTIVE: This paper presents the conceptual framework and research design of the chronic NCD risk factor surveillance within nine rural INDEPTH HDSS settings in Asia. METHODS: This multi-site study was designed as a baseline cross-sectional survey with sufficient sample size to measure trends over time. In each of nine HDSS sites in five Asian countries, a sample of 2,000 men and women aged 25-64 years, using the WHO STEPwise approach to Surveillance (http://who.int/chp/steps), was selected using stratified random sampling (in each 10-year interval) from the HDSS sampling frame. RESULTS: A total of 18,494 men and women from the nine sites were interviewed with an overall response rate of 98%. The major NCDs risk factors included self-reported information on tobacco and alcohol consumption, fruit and vegetable intake, physical activity patterns, and measured body weight, height, waist circumference, and blood pressure. A series of training sessions were conducted for research scientists, supervisors, and surveyors in each site. Data quality was ensured through spot check, re check, and data validation procedures, including accuracy and completeness of data obtained. Standardised data entry programme, created using the EPIDATA software, was used to ensure uniform database structure across sites. The data merging and analysis were done using STATA Version 10. CONCLUSION: This multi site study confirmed the feasibility of conducting chronic NCD risk factor surveillance in the low and middle-income settings by integrating the chronic NCDs risk factor surveillance into an existing HDSS data collection and management setting. This collaborative work has provided reliable epidemiological data as a basis for developing chronic NCD prevention and control activities. PMID- 20027264 TI - Climate change, direct heat exposure, health and well-being in low and middle income countries. PMID- 20027265 TI - Invited Commentary to the paper 'Dying to count: mortality surveillance in resource-poor countries' by Edward Fottrell. PMID- 20027266 TI - Willingness to volunteer in a Phase I/II HIV vaccine trial: a study among police officers in Dar es Salaam, Tanzania. AB - BACKGROUND: As HIV infection continues to devastate low-income countries, efforts to search for an effective HIV vaccine are crucial. Therefore, participation in HIV vaccine trials will be useful for the development of a preventive vaccine that will work and thus reduce the global HIV epidemic. OBJECTIVE: The objective of this study was to analyse the willingness to volunteer (WTV) in a Phase I/II HIV vaccine trial among police officers in Dar es Salaam, Tanzania. DESIGN: We included a convenience sample of 329 participants (79% males) from sensitisation workshops that were held once at each of the 32 police stations. Participants were recruited from 23 stations which were included according to availability. Data about personal characteristics, general HIV and AIDS knowledge and sexual behaviour, attitudes towards vaccines and willingness to participate in the HIV vaccine trial were obtained through an interview-administered questionnaire with both closed and open-ended questions. RESULTS: Overall, 61% of the participants expressed WTV in HIV vaccine trials. WTV was significantly associated with: positive attitude towards use of effective vaccine, Odds ratio (OR), 36.48 (95% CI: 15.07-88.28); the intention to tell others about one's decision to participate in the trial, OR, 6.61 (95% CI: 3.89-11.24); Tanzania becoming a partner in developing the vaccine, OR, 4.28 (95% CI: 2.28-8.03); having an extra sexual partner, OR, 3.05 (95% CI: 1.63-5.69); perceived higher risk of getting HIV infection, OR, 2.11 (95% CI: 1.34-3.33); and high knowledge about HIV and AIDS, OR, 1.92 (95% CI: 1.22-3.01). CONCLUSION: The results indicated that a majority of police officers in this study were willing to participate in HIV vaccine trials. However, there is a need to provide the respondents with precise information about the purpose of a Phase I/II HIV vaccine trial and the fact that it does not protect against HIV infection, in order to avoid increasing risky behaviour. PMID- 20027267 TI - Towards reduction of maternal and perinatal mortality in rural Burkina Faso: communities are not empty vessels. AB - BACKGROUND: Reducing maternal and perinatal mortality in sub Saharan Africa remains challenging and requires effective and context specific interventions. OBJECTIVE: The aims of this paper were to demonstrate the impact of the community mobilisation of the Skilled Care Initiative (SCI) in reducing maternal and perinatal mortality and to describe the concept and implementation in order to guide replication and scaling up. DESIGNS: A quasi experimental design was used to assess the extent to which the SCI was associated with increased institutional births, maternal and perinatal mortality reduction in an intervention (Ouargaye) versus a comparison (Diapaga) district. A geo-referenced census was conducted to retrospectively assess changes in outcomes and process measures. A detailed description of activities, rationale and timing of implementation were gathered from the SCI project officers and summarised. Data analyses included descriptive statistics and multivariate analyses. RESULTS: At macro level, the main significant difference between Ouargaye and Diapaga districts was the scope and intensity of the community-based interventions implemented in Ouargaye. There was a temporal association relationship before and after the implementation of the demand-driven interventions and a remarkable 30% increase in institutional births in the intervention district compared to 10% increase in comparison district. There was a significant reduction of perinatal mortality rates (OR =0.75, CI 0.70 0.80) in intervention district and a larger decrease in maternal mortality ratios in intervention district, although statistical significance was not reached. A comprehensive framework of community mobilisation strategy is proposed to improve maternal and child health in poorest communities. CONCLUSION: Controlling for the availability and quality of health services, working in partnership and effectively with communities, and not for them - hence characterising communities as not being empty vessels - can have impacts on outcomes. Here, in the district with a community mobilisation programme, there was a marked increase in institutional births and reductions in maternal and perinatal deaths. PMID- 20027268 TI - Survey of ENT services in Africa: need for a comprehensive intervention. AB - BACKGROUND: Burden of disease (BOD) is greatest in resource-starved regions such as Africa. Even though hearing disability ranks third on the list of non-fatal disabling conditions in low- and middle-income countries, ear, nose, and throat (ENT) disorders are not specifically coded for within the framework governing global BOD estimates, and in discussions about health challenges, non communicable diseases receive scant attention. Implementing cost-effective interventions to address conditions largely neglected by global estimates of BOD such as hearing loss are important contributors to health and economic development. OBJECTIVES: Establish a database of ENT, audiology, and speech therapy services in Sub-Saharan Africa; create awareness about the status of these services; propose effective intervention; gather data to lobby African governments, donor countries, and aid organizations; determine need for Developing World Forum for ENT, Audiology, and Speech Therapy services. DESIGN: Survey of ENT, audiology, and speech therapy services and training in 18 Sub Saharan Africa countries. RESULTS: There is an alarming paucity of services and training opportunities, and there is a large gap between higher technology, expensive health care in high-income countries and lower technology, low-cost practice in low-income countries. CONCLUSIONS: Lower technology and lower cost Developing World medical practice should be recognized and fostered as a field of medical practice, teaching, and research. Developing World centers of excellence must be fostered to take a lead in teaching, training, and research. A Developing World Forum for ENT Surgery, Audiology, and Speech Therapy, directed and driven by Africa and the Developing World, supported by the First World, should be established, to develop a comprehensive intervention to turn around the severe shortage of services and expertise in the Developing World. Global health policies and practices should include new norms and standards which serve the interests of the global community, and are based on current realities of global health. PMID- 20027269 TI - Dying to count: mortality surveillance in resource-poor settings. AB - Reliable cause-specific mortality data constitute a crucial resource for health monitoring, service planning and prioritisation. However, in the majority of the world's poorest settings, systematic health and vital event surveillance systems are weak or non-existent. As such, deaths are not counted and causes of death remain unregistered for more than two-thirds of the world's population.For researchers, health workers and policy makers in resource-poor settings, therefore, attempts to measure mortality have to be implemented from first principles. As a result, there is wide variation in mortality surveillance methodologies in different settings, and lack of standardisation and rigorous validation of these methods hinder meaningful comparison of mortality data between settings and over time.With a particular focus on Health and Demographic Surveillance Systems (HDSSs), this paper summarises recent research and conceptual development of certain methodological aspects of mortality surveillance stemming from a series of empirical investigations. The paper describes the advantages and limitations of various methods in particular contexts, and argues that there is no single methodology to satisfy all data needs. Rather, methodological decisions about mortality measurement should be a synthesis of all available knowledge relating to clearly defined concepts of why data are being collected, how they can be used and when they are of good enough quality to inform public health action. PMID- 20027270 TI - Associations between urbanisation and components of the health-risk transition in Thailand. A descriptive study of 87,000 Thai adults. AB - BACKGROUND: Social and environmental changes have accompanied the ongoing rapid urbanisation in a number of countries during recent decades. Understanding of its role in the health-risk transition is important for health policy development at national and local level. Thailand is one country facing many of the health challenges of urbanisation. OBJECTIVE: To identify potential associations between individual migration between rural and urban areas and exposure to specific social, economic, environmental and behavioural health determinants. DESIGN: Baseline data from a cohort of 87,134 Thai open university students surveyed in 2005 (mean age 31 years). Four urbanisation status groups were defined according to self-reported location of residence (rural: R or urban: U) in 2005 and when the respondent was 10-12 years old (yo). RESULTS: Fourty-four percent were living in rural areas in 2005 and when they were 10-12yo (Group RR: ruralites); 20% always lived in urban areas (UU: urbanites); 32% moved from rural to urban areas (RU: urbanisers); 4% moved in the other direction (UR: de-urbanisers). The ruralites and urbanites often were the two extremes, with the urbanisers maintaining some of the determinants patterns from ruralites and the de urbanisers maintaining patterns from urbanites. There was a strong relationship between urbanisation status, from RR to RU to UR to UU, and personal income, availability of modern home appliances, car ownership, consumption of 'junk food' and physical inactivity. Urbanisers reported worse socio-environmental conditions and worse working conditions than the other groups. De-urbanisers had the highest rates of smoking and drinking. CONCLUSIONS: An urbanisation measure derived from self-reported location of residence gave new insights into the health risk exposures of migrants relative to permanent rural and permanent urban dwellers. Living in urban areas is an important upstream determinant of health in Thailand and urbanisation is a key element of the Thai health-risk transition. PMID- 20027271 TI - Decreasing childhood mortality and increasing proportion of malaria deaths in rural Burkina Faso. AB - BACKGROUND: Malaria is the leading cause of death among children less than five years of age in sub-Saharan Africa (SSA), however, precise estimates on the burden of malaria are lacking. The aim of this study was to describe temporal trends for malaria and all-cause mortality by combining a series of clinical and intervention studies conducted in Burkina Faso. METHODS: Data from a demographic surveillance system was used to follow-up children under five years who participated in five observational and intervention studies between June 1999 and December 2004 in rural north-western Burkina Faso. Mortality data was analyzed with cause-specific mortality ascertained using the verbal autopsy method. Person years (PY) of observations were computed and age-standardized mortality rates (MR) for all-causes and malaria (adjusted for missing causes of death) were calculated. Rate ratios to investigate mortality variations over years were calculated using multivariate Poisson regression. RESULTS: The study followed 6,387 children aged less than five years (mean follow-up: 2.8 years; 16,099 PY). During the study period, 443 deaths were registered with malaria accounting for 49% of all deaths. All-cause and malaria-specific MR were 26.7 (95% CI: 24.2 29.2) and 15.8 (95% CI: 14.217.7) per 1,000 PY. All-cause MR declined over years of follow-up (from 31.2 to 16.3 per 1,000 PY in 1999/2000 to 2004, respectively) but malaria MR remained relatively stable (from 15.8 to 12.1 per 1,000 PY in 1999/2000 to 2004, respectively) resulting in an increasing relative effect of malaria on all-cause mortality. Variations in all-cause and malaria-specific mortality were observed with increasing age and across village town clusters. CONCLUSION: The findings of this study support the continuously decreasing trend of all-cause mortality in most of SSA, but call for more efforts to comprehensively address malaria with existing control tools such as insecticide treated bed nets and effective first-line combination therapies. PMID- 20027272 TI - A practical approach to identifying maternal deaths missed from routine hospital reports: lessons from Indonesia. AB - BACKGROUND: Accurate estimates of the number of maternal deaths in both the community and facility are important, in order to allocate adequate resources to address such deaths. On the other hand, current studies show that routine methods of identifying maternal deaths in facilities underestimate the number by more than one-half. OBJECTIVE: To assess the utility of a new approach to identifying maternal deaths in hospitals. METHOD: Deaths of women of reproductive age were retrospectively identified from registers in two district hospitals in Indonesia over a 24-month period. Based on information retrieved, deaths were classified as 'maternal' or 'non-maternal' where possible. For deaths that remained unclassified, a detailed case note review was undertaken and the extracted data were used to facilitate classification. RESULTS: One hundred and fifty-five maternal deaths were identified, mainly from the register review. Only 67 maternal deaths were recorded in the hospitals' routine reports over the same period. This underestimation of maternal deaths was partly due to the incomplete coverage of the routine reporting system; however, even in the wards where routine reports were made, the study identified twice as many deaths. CONCLUSION: The RAPID method is a practical method that provides a more complete estimate of hospital maternal mortality than routine reporting systems. PMID- 20027273 TI - Epidemiological transition in rural Bangladesh, 1986-2006. AB - BACKGROUND: For understanding epidemiological transition, Health and Demographic Surveillance System plays an important role in developing and resource-constraint setup where accurate information on vital events (e.g. births, deaths) and cause of death is not available. METHODS: This study aimed to assess existing level and trend of causes of 18,917 deaths in Matlab, a rural area of Bangladesh, during 1986-2006 and to project future scenarios for selected major causes of death. RESULTS: The results demonstrated that Matlab experienced a massive change in the mortality profile from acute, infectious, and parasitic diseases to non communicable, degenerative, and chronic diseases during the last 20 years. It also showed that over the period 1986-2006, age-standardized mortality rate (for both sexes) due to diarrhea and dysentery reduced by 86%, respiratory infections by 79%, except for tuberculosis which increased by 173%. On the other hand, during the same period, mortality due to cardiovascular and cerebrovascular diseases increased by a massive 3,527% and malignant neoplasms by 495%, whereas mortality due to chronic obstructive pulmonary disease and injury remained in the similar level (12-13% increase). CONCLUSION: The trend of selected causes of death demonstrates that in next two decades, deaths due to communicable diseases will decline substantially and the mortality due to non-communicable diseases (NCDs) will increase at massive proportions. Despite Matlab's significant advances in socio-demographic indicators, emergence of NCDs and mortality associated with it would be the major cause for concern in the coming years. PMID- 20027274 TI - The implications of policy changes on the uptake of a PMTCT programme in rural Malawi: first three years of experience. AB - OBJECTIVE: To study the implications of policy changes on the demand for antenatal care (ANC), HIV testing and hospital delivery among pregnant women in rural Malawi. DESIGN: Retrospective analysis of monthly reports. SETTING: Malamulo SDA hospital in Thyolo district, Makwasa, Malawi. METHODS: Three hospital-based registers were analysed from 2005 to 2007. These were general ANC, delivery and Prevention of Mother to Child Transmission (PMTCT) registers. Observations were documented regarding the introduction of specific policies and when changes were effected. Descriptive analytical methods were used. RESULTS: The ANC programme reached 4,528 pregnant mothers during the study period. HIV testing among the ANC attendees increased from 52.6 to 98.8% after the introduction of routine (opt-out) HIV testing and 15.6% of them tested positive. After the introduction of free maternity services, ANC attendance increased by 42% and the ratio of hospital deliveries to ANC attendees increased from 0.50:1 to 0.66:1. Of the HIV-tested ANC attendees, 52.6% who tested positive delivered in the hospital and got nevirapine at the time of delivery. CONCLUSIONS: Increasing maternity service availability and uptake can increase the coverage of PMTCT programmes. Barriers such as economic constraints that prevent women in poor communities from accessing services can be removed by making maternity services free. However, it is likely, particularly in resource-poor settings, that significant increases in PMTCT coverage among those at risk can only be achieved by substantially increasing uptake of general ANC and delivery services. PMID- 20027275 TI - Providing free maternal health care: ten lessons from an evaluation of the national delivery exemption policy in Ghana. AB - BACKGROUND: There is a growing movement, globally and in the Africa region, to reduce financial barriers to health care generally, but with particular emphasis on high priority services and vulnerable groups. OBJECTIVE: This article reports on the experience of implementing a national policy to exempt women from paying for delivery care in public, mission and private health facilities in Ghana. DESIGN: Using data from a complex evaluation which was carried out in 2005-2006, lessons are drawn which can inform other countries starting or planning to implement similar service-based exemption policies. RESULTS: On the positive side, the experience of Ghana suggests that delivery exemptions can be effective and cost-effective, and that despite being universal in application, they can benefit the poor. However, certain 'negative' lessons are also drawn from the Ghana case study, particularly on the need for adequate funding, and for strong institutional ownership. It is also important to monitor the financial transfers which reach households, to ensure that providers are passing on benefits in full, while being adequately reimbursed themselves for their loss of revenue. Careful consideration should also be given to staff motivation and the role of different providers, as well as quality of care constraints, when designing the exemptions policy. All of this should be supported by a proactive approach to monitoring and evaluation. CONCLUSION: The recent movement towards making delivery care free to all women is a bold and timely action which is supported by evidence from within and beyond Ghana. However, the potential for this to translate into reduced mortality for mothers and babies fundamentally depends on the effectiveness of its implementation. PMID- 20027276 TI - Financial incentives to influence maternal mortality in a low-income setting: making available 'money to transport' - experiences from Amarpatan, India. AB - OBJECTIVES: Only 40.7% women in India deliver in an institution; leaving many vulnerable to maternal morbidity and mortality (India has 22% of global maternal deaths). While limited accessibility to functioning institutions may account in part, a common reason why women deliver at home is poverty. A lack of readily available financial resources for families to draw upon at the time of labor to transport the mother to an institution, is often observed. This paper reports a yearlong collaborative intervention (between the University and Department of Health) to study if providing readily available and easily accessible funds for emergency transportation would reduce maternal deaths in a rural, low income, and high maternal mortality setting in central India. It aimed to obviate a deterrent to emergency obstetric care; the non-availability of resources with mothers when most needed. Issues in implementation are also discussed. METHODS: Maternal deaths were actively identified in block Amarpatan (0.2 million population) over a 2-year period. The project, with participation from local government and other groups, trained 482 local health care providers (public and private) to provide antenatal care. Emergency transport money (in cash) was placed with one provider in each village. Maternal mortality in the adjacent block (Maihar) was followed (as a 'control' block). RESULTS: Maternal deaths in Amarpatan decreased during the project year relative to the previous year, or in the control block the same year. DISCUSSION AND CONCLUSIONS: Issues in implementation of the cash incentive scheme are discussed. Although the intervention reduced maternal deaths in this low-income setting, chronic poverty and malnutrition are underlying structural problems that need to be addressed. PMID- 20027277 TI - Knowledge, attitudes and practice about malaria in rural Tigray, Ethiopia. AB - OBJECTIVE: To assess the knowledge, attitude and practice (KAP) regarding malaria and their determinants in a rural population of northern Ethiopia. METHODS: The study was conducted in the district of Samre Saharti, Tigray, northern Ethiopia. A structured questionnaire collecting socio-demographic and malaria-related KAP information was administered to the mothers from a representative sample of households. RESULTS: A total of 1652 questionnaires were available for analysis. Most of the respondents (92.7%) were able to mention at least one symptom of malaria. Mosquito as a cause of malaria was recognized by nearly half of the respondents (48.8%). Most of the households had a bed net (85.9%). To have a literate person at home, to belong to the lowland stratum, to have received some type of health education and to own a radio were associated with the knowledge of malaria. A strong association remained between living in the lowland stratum, to own a radio and to live close to the health post and the use of ITN. Being a housewife, lack of health education and to live further than 60 minutes walking distance to the health post were related to a delay on treatment finding. CONCLUSION: This study has identified some aspects which the MCP might need to improve. The knowledge about malaria transmission should be strengthened. Promotion of literacy and participation in health education are vital components in terms of malaria knowledge and practice. Issues related to geographical location and accessibility to health post should be also carefully examined. PMID- 20027278 TI - Adolescent medicine: attitudes, training, and experience of pediatric, family medicine, and obstetric-gynecology residents. AB - PURPOSE: Several studies have documented a deficiency in the delivery of preventive services to adolescents during physician visits in the United States. This study sought to assess and compare pediatric, family medicine (FM), and obstetrics and gynecology (OB/GYN) resident perceptions of their responsibility, training, and experience with providing comprehensive health care services to adolescents. METHODS: A 57-item, close-ended survey was designed and administered to assess resident perceptions of the scope of their practice, training, and experience with providing adolescent health care across a series of health care categories. RESULTS: Of the 87 respondents (31 OB/GYN, 29 FM, and 27 pediatric), most residents from all three fields felt that the full range of adolescent preventive and clinical services represented in the survey fell under their scope of practice. Residents from all three fields need more training and experience with mental health issues, referring teenagers to substance abuse treatment programs, and addressing physical and sexual abuse. In addition, OB-GYN residents reported deficiencies in training and experience regarding several preventive counseling and general health services, while pediatric residents reported deficiencies in training and experience regarding sexual health services. CONCLUSIONS: Our results indicate that at this time, residents from these three specialties are not optimally prepared to provide the full range of recommended preventive and clinical services to adolescents. PMID- 20027279 TI - Pandemic influenza H1N1 2009, innate immunity, and the impact of immunosenescence on influenza vaccine. AB - Seasonal and pandemic strains of influenza have widespread implications for the global economy and global health. This has been highlighted recently as the epidemiologic characteristics for hospitalization and mortality for pandemic influenza H1N1 2009 are now emerging. While treatment with neuraminidase inhibitors are effective for seasonal and pandemic influenza, prevention of morbidity and mortality through effective vaccines requires a rigorous process of research and development. Vulnerable populations such as older adults (i.e., > age 65 years) suffer the greatest impact from seasonal influenza yet do not have a consistent seroprotective response to seasonal influenza vaccines due to a combination of factors. This short narrative review will highlight the emerging epidemiologic characteristics of pandemic H1N1 2009 and focus on immunosenescence, innate immune system responses to influenza virus infection and vaccination, and influenza vaccine responsiveness as it relates to seasonal and H1N1 pandemic influenza vaccines. PMID- 20027281 TI - Changing anthropology, changing society. AB - Fifty years after the founding of the field of medical anthropology, the Society for Medical Anthropology of the American Anthropological Association held its first independent meeting on September 24-27, 2009, at Yale University. PMID- 20027280 TI - Influenza A: understanding the viral life cycle. AB - Influenza A virus belongs to the family of Orthomyxoviridae. It is an enveloped virus with a negative sense RNA segmented genome that encodes for 11 viral genes. This virus has evolved a number of mechanisms that enable it to invade host cells and subvert the host cell machinery for its own purpose, that is, for the sole production of more virus. Two of the mechanisms that the virus uses are "cap snatching" and preventing the host cell from expressing its own genes. This mini review provides a brief overview as to how the virus is able to invade host cells, replicate itself, and exit the host cell. PMID- 20027282 TI - The intersection of anthropology and medicine as portrayed in Paul Farmer's photo album. AB - Paul Farmer, physician, anthropologist, and author, spoke at the 2009 Society for Medical Anthropology Conference at Yale University in September. PMID- 20027283 TI - Genetics and genomics as an anthropological subject: approaching with caution. AB - Speaking at the 2009 Society for Medical Anthropology Conference, cultural anthropologist Margaret Lock pointed out that the advent of the genomic revolution brings with it important societal, political, and social issues that have the potential to radically change both human life and interaction. PMID- 20027284 TI - From race to repositories: bioethics in a genomics age. AB - Medical anthropologist Barbara Koenig spoke on the intersection of bioethics and genomics at the 2009 Society for Medical Anthropology Conference at Yale University in New Haven, Connecticut. PMID- 20027285 TI - Reflections on the future of anthropology. AB - In his plenary session entitled Five Questions on the Future, Harvard anthropologist Arthur Kleinman capitalized on the 2009 Society for Medical Anthropology Conference's theme of Medical Anthropology at the Intersections to speculate on the future of the discipline. PMID- 20027286 TI - Global public health through the eyes of medical anthropologist Didier Fassin. AB - At the 2009 Society for Medical Anthropology Conference at Yale University, anthropologist Didier Fassin discussed social inequality and the politicization of health in the context of global public health. PMID- 20027287 TI - Training, communication, and competence: the making of health care professionals. AB - The role of medical anthropology in tackling the problems and challenges at the intersections of public health, medicine, and technology was addressed during the 2009 Society for Medical Anthropology Conference at Yale University in an interdisciplinary panel session entitled Training, Communication, and Competence: The Making of Health Care Professionals. PMID- 20027288 TI - Allergen challenge induces Ifng dependent GTPases in the lungs as part of a Th1 transcriptome response in a murine model of allergic asthma. AB - According to the current paradigm, allergic airway inflammation is mediated by Th2 cytokines and pro-inflammatory chemokines. Since allergic inflammation is self-limited, we hypothesized that allergen challenge simultaneously induces anti inflammatory genes to counter-balance the effects of Th2 cytokines and chemokines. To identify these putative anti-inflammatory genes, we compared the gene expression profile in the lungs of ragweed-sensitized mice four hours after challenge with either PBS or ragweed extract (RWE) using a micro-array platform. Consistent with our hypothesis, RWE challenge concurrently upregulated Th1 associated early target genes of the Il12/Stat4 pathway, such as p47 and p65 GTPases (Iigp, Tgtp and Gbp1), Socs1, Cxcl9, Cxcl10 and Gadd45g with the Th2 genes Il4, Il5, Ccl2 and Ccl7. These Th1-associated genes remain upregulated longer than the Th2 genes. Augmentation of the local Th1 milieu by administration of Il12 or CpG prior to RWE challenge further upregulated these Th1 genes. Abolition of the Th1 response by disrupting the Ifng gene increased allergic airway inflammation and abrogated RWE challenge-induced upregulation of GTPases, Cxcl9, Cxcl10 and Socs1, but not Gadd45g. Our data demonstrate that allergen challenge induces two sets of Th1-associated genes in the lungs: 1) Ifng dependent genes such as p47 and p65 GTPases, Socs1, Cxcl9 and Cxcl10 and 2) Ifng independent Th1-inducing genes like Gadd45g. We propose that allergen-induced airway inflammation is regulated by simultaneous upregulation of Th1 and Th2 genes, and that persistent unopposed upregulation of Th1 genes resolves allergic inflammation. PMID- 20027289 TI - Is the scale up of malaria intervention coverage also achieving equity? AB - BACKGROUND AND METHODS: Malaria in Africa is most severe in young children and pregnant women, particularly in rural and poor households. In many countries, malaria intervention coverage rates have increased as a result of scale up; but this may mask limited coverage in these highest-risk populations. Reports were reviewed from nationally representative surveys in African malaria-endemic countries from 2006 through 2008 to understand how reported intervention coverage rates reflect access by the most at-risk populations. RESULTS: Reports were available from 27 Demographic and Health Surveys (DHSs), Multiple Indicator Cluster Surveys (MICSs), and Malaria Indicator Surveys (MISs) during this interval with data on household intervention coverage by urban or rural setting, wealth quintile, and sex. Household ownership of insecticide-treated mosquito nets (ITNs) varied from 5% to greater than 60%, and was equitable by urban/rural and wealth quintile status among 13 (52%) of 25 countries. Malaria treatment rates for febrile children under five years of age varied from less than 10% to greater than 70%, and while equitable coverage was achieved in 8 (30%) of 27 countries, rates were generally higher in urban and richest quintile households. Use of intermittent preventive treatment in pregnant women varied from 2% to more than 60%, and again tended to be higher in urban and richest quintile households. Across all countries, there were no significant male/female inequalities seen for children sleeping under ITNs or receiving antimalarial treatment for febrile illness. Parasitemia and anemia rates from eight national surveys showed predominance in poor and rural populations. CONCLUSIONS/SIGNIFICANCE: Recent efforts to scale up malaria intervention coverage have achieved equity in some countries (especially with ITNs), but delivery methods in other countries are not addressing the most at-risk populations. As countries seek universal malaria intervention coverage, their delivery systems must reach the rural and poor populations; this is not a small task, but it has been achieved in some countries. PMID- 20027290 TI - Tandem ChoRE and CCAAT motifs and associated factors regulate Txnip expression in response to glucose or adenosine-containing molecules. AB - BACKGROUND: Thioredoxin interacting protein (Txnip) is a multifunctional protein involved in regulation of cell cycle events and cellular metabolism. The expression of Txnip is known to be induced by glucose, adenosine-containing molecules, and other physiological cues; however, the underlying regulatory mechanisms remain elusive. METHODOLOGY/PRINCIPAL FINDINGS: In this study, using promoter reporter, electrophoresis mobility shift (EMSA), and chromatin immuno precipitation (ChIP) assays, we have identified an additional carbohydrate response element (ChoRE) on the promoter of Txnip gene, which functions cooperatively with the earlier identified ChoRE to mediate optimal Txnip expression. However, these two ChoREs are not sufficient to mediate the induction of Txnip expression by glucose or adenosine-containing molecules; and two CCAAT boxes, both of which can recruit nuclear factor Y (NF-Y) to the Txnip promoter, are also required for the induction. Accordingly, we have found that the function of ChoREs and associated factors is contingent on tandem CCAAT boxes, in that occupancy of the Txnip promoter by NF-Y is a prerequisite for efficacious recruitment of Mondo/MLX to ChoREs under glucose stimulation. CONCLUSIONS/SIGNIFICANCE: Our findings suggest a synergy between the tandem CCAAT and ChoRE motifs and associated NF-Y and Mondo/MLX transcription factors in enhancing transcription from the Txnip promoter. This piece of information will be helpful for future dissection of molecular mechanisms governing the transcriptional regulation of Txnip, a glucose responsive gene. PMID- 20027291 TI - Salmonella induced IL-23 and IL-1beta allow for IL-12 production by monocytes and Mphi1 through induction of IFN-gamma in CD56 NK/NK-like T cells. AB - BACKGROUND: The type-1 cytokine pathway plays a pivotal role in immunity against intracellular bacterial pathogens such as Salmonellae and Mycobacteria. Bacterial stimulation of pattern recognition receptors on monocytes, macrophages and dendritic cells initiates this pathway, and results in the production of cytokines that activate lymphocytes to produce interferon (IFN)-gamma. Interleukin (IL)-12 and IL-23 are thought to be the key cytokines required for initiating a type-1 cytokine immune response to Mycobacteria and Salmonellae. The relative contribution of IL-23 and IL-12 to this process is uncertain. METHODOLOGY/PRINCIPAL FINDINGS: We show that various TLR agonists induce the production of IL-23 but not IL-12 in freshly isolated human monocytes and cultured human macrophages. In addition, type 1 pro-inflammatory macrophages (Mphi1) differentiated in the presence of GM-CSF and infected with live Salmonella produce IL-23, IL-1beta and IL-18, but not IL-12. Supernatants of Salmonella-infected Mphi1 contained more IL-18 and IL-1beta as compared with supernatants of Mphi1 stimulated with isolated TLR agonists, and induced IFN gamma production in human CD56(+) cells in an IL-23 and IL-1beta-dependent but IL 12-independent manner. In addition, IL-23 together with IL-18 or IL-1beta led to the production of GM-CSF in CD56(+) cells. Both IFN-gamma and GM-CSF enhanced IL 23 production by monocytes in response to TLR agonists, as well as induced IL-12 production. CONCLUSIONS/SIGNIFICANCE: The findings implicate a positive feedback loop in which IL-23 can enhance its release via induction of IFN-gamma and GM CSF. The IL-23 induced cytokines allow for the subsequent production of IL-12 and amplify the IFN-gamma production in the type-1 cytokine pathway. PMID- 20027293 TI - Breaking the waves: modelling the potential impact of public health measures to defer the epidemic peak of novel influenza A/H1N1. AB - BACKGROUND: On June 11, 2009, the World Health Organization declared phase 6 of the novel influenza A/H1N1 pandemic. Although by the end of September 2009, the novel virus had been reported from all continents, the impact in most countries of the northern hemisphere has been limited. The return of the virus in a second wave would encounter populations that are still nonimmune and not vaccinated yet. We modelled the effect of control strategies to reduce the spread with the goal to defer the epidemic wave in a country where it is detected in a very early stage. METHODOLOGY/PRINCIPAL FINDINGS: We constructed a deterministic SEIR model using the age distribution and size of the population of Germany based on the observed number of imported cases and the early findings for the epidemiologic characteristics described by Fraser (Science, 2009). We propose a two-step control strategy with an initial effort to trace, quarantine, and selectively give prophylactic treatment to contacts of the first 100 to 500 cases. In the second step, the same measures are focused on the households of the next 5,000 to 10,000 cases. As a result, the peak of the epidemic could be delayed up to 7.6 weeks if up to 30% of cases are detected. However, the cumulative attack rates would not change. Necessary doses of antivirals would be less than the number of treatment courses for 0.1% of the population. In a sensitivity analysis, both case detection rate and the variation of R0 have major effects on the resulting delay. CONCLUSIONS/SIGNIFICANCE: Control strategies that reduce the spread of the disease during the early phase of a pandemic wave may lead to a substantial delay of the epidemic. Since prophylactic treatment is only offered to the contacts of the first 10,000 cases, the amount of antivirals needed is still very limited. PMID- 20027292 TI - A protocadherin-cadherin-FLRT3 complex controls cell adhesion and morphogenesis. AB - BACKGROUND: Paraxial protocadherin (PAPC) and fibronectin leucine-rich domain transmembrane protein-3 (FLRT3) are induced by TGFbeta signaling in Xenopus embryos and both regulate morphogenesis by inhibiting C-cadherin mediated cell adhesion. PRINCIPAL FINDINGS: We have investigated the functional and physical relationships between PAPC, FLRT3, and C-cadherin. Although neither PAPC nor FLRT3 are required for each other to regulate C-cadherin adhesion, they do interact functionally and physically, and they form a complex with cadherins. By itself PAPC reduces cell adhesion physiologically to induce cell sorting, while FLRT3 disrupts adhesion excessively to cause cell dissociation. However, when expressed together PAPC limits the cell dissociating and tissue disrupting activity of FLRT3 to make it effective in physiological cell sorting. PAPC counteracts FLRT3 function by inhibiting the recruitment of the GTPase RND1 to the FLRT3 cytoplasmic domain. CONCLUSIONS/SIGNIFICANCE: PAPC and FLRT3 form a functional complex with cadherins and PAPC functions as a molecular "governor" to maintain FLRT3 activity at the optimal level for physiological regulation of C cadherin adhesion, cell sorting, and morphogenesis. PMID- 20027294 TI - Validation of inverse seasonal peak mortality in medieval plagues, including the Black Death, in comparison to modern Yersinia pestis-variant diseases. AB - BACKGROUND: Recent studies have noted myriad qualitative and quantitative inconsistencies between the medieval Black Death (and subsequent "plagues") and modern empirical Y. pestis plague data, most of which is derived from the Indian and Chinese plague outbreaks of A.D. 1900+/-15 years. Previous works have noted apparent differences in seasonal mortality peaks during Black Death outbreaks versus peaks of bubonic and pneumonic plagues attributed to Y. pestis infection, but have not provided spatiotemporal statistical support. Our objective here was to validate individual observations of this seasonal discrepancy in peak mortality between historical epidemics and modern empirical data. METHODOLOGY/PRINCIPAL FINDINGS: We compiled and aggregated multiple daily, weekly and monthly datasets of both Y. pestis plague epidemics and suspected Black Death epidemics to compare seasonal differences in mortality peaks at a monthly resolution. Statistical and time series analyses of the epidemic data indicate that a seasonal inversion in peak mortality does exist between known Y. pestis plague and suspected Black Death epidemics. We provide possible explanations for this seasonal inversion. CONCLUSIONS/SIGNIFICANCE: These results add further evidence of inconsistency between historical plagues, including the Black Death, and our current understanding of Y. pestis-variant disease. We expect that the line of inquiry into the disputed cause of the greatest recorded epidemic will continue to intensify. Given the rapid pace of environmental change in the modern world, it is crucial that we understand past lethal outbreaks as fully as possible in order to prepare for future deadly pandemics. PMID- 20027295 TI - Fatty acid synthase impacts the pathobiology of Candida parapsilosis in vitro and during mammalian infection. AB - Cytosolic fungal fatty acid synthase is composed of two subunits alpha and beta, which are encoded by Fas1 and Fas2 genes. In this study, the Fas2 genes of the human pathogen Candida parapsilosis were deleted using a modified SAT1 flipper technique. CpFas2 was essential in media lacking exogenous fatty acids and the growth of Fas2 disruptants (Fas2 KO) was regulated by the supplementation of different long chain fatty acids, such as myristic acid (14ratio0), palmitic acid (16ratio0), and Tween 80, in a dose-specific manner. Lipidomic analysis revealed that Fas2 KO cells were severely restricted in production of unsaturated fatty acids. The Fas2 KO strains were unable to form normal biofilms and were more efficiently killed by murine-like macrophages, J774.16, than the wild type, heterozygous and reconstituted strains. Furthermore, Fas2 KO yeast were significantly less virulent in a systemic murine infection model. The Fas2 KO cells were also hypersensitive to human serum, and inhibition of CpFas2 in WT C. parapsilosis by cerulenin significantly decreased fungal growth in human serum. This study demonstrates that CpFas2 is essential for C. parapsilosis growth in the absence of exogenous fatty acids, is involved in unsaturated fatty acid production, influences fungal virulence, and represents a promising antifungal drug target. PMID- 20027296 TI - Adult rat bones maintain distinct regionalized expression of markers associated with their development. AB - The incidence of limb bone fracture and subsequent morbidity and mortality due to excessive bone loss is increasing in the progressively ageing populations of both men and women. In contrast to bone loss in the weight-bearing limb, bone mass in the protective skull vault is maintained. One explanation for this could be anatomically diverse bone matrix characteristics generated by heterogeneous osteoblast populations. We have tested the hypothesis that adult bones demonstrate site-specific characteristics, and report differences at the organ, cell and transcriptome levels. Limb bones contain greater amounts of polysulphated glycosaminoglycan stained with Alcian Blue and have significantly higher osteocyte densities than skull bone. Site-specific patterns persist in cultured adult bone-derived cells both phenotypically (proliferation rate, response to estrogen and cell volumes), and at the level of specific gene expression (collagen triple helix repeat containing 1, reelin and ras-like and estrogen-regulated growth inhibitor). Based on genome-wide mRNA expression and cluster analysis, we demonstrate that bones and cultured adult bone-derived cells segregate according to site of derivation. We also find the differential expression of genes associated with embryological development (Skull: Zic, Dlx, Irx, Twist1 and Cart1; Limb: Hox, Shox2, and Tbx genes) in both adult bones and isolated adult bone-derived cells. Together, these site-specific differences support the view that, analogous to different muscle types (cardiac, smooth and skeletal), skull and limb bones represent separate classes of bone. We assign these differences, not to mode of primary ossification, but to the embryological cell lineage; the basis and implications of this division are discussed. PMID- 20027297 TI - The constitutively active V2 receptor mutants conferring NSIAD are weakly sensitive to agonist and antagonist regulation. AB - Patients having the nephrogenic syndrome of inappropriate antidiuresis present either the R137C or R137L V2 mutated receptor. While the clinical features have been characterized, the molecular mechanisms of functioning of these two mutants remain elusive. In the present study, we compare the pharmacological properties of R137C and R137L mutants with the wild-type and the V2 D136A receptor, the latter being reported as a highly constitutively active receptor. We have performed binding studies, second messenger measurements and BRET experiments in order to evaluate the affinities of the ligands, their agonist and antagonist properties and the ability of the receptors to recruit beta-arrestins, respectively. The R137C and R137L receptors exhibit small constitutive activities regarding the G(s) protein activation. In addition, these two mutants induce a constitutive beta-arrestin recruitment. Of interest, they also exhibit weak sensitivities to agonist and to inverse agonist in term of G(s) protein coupling and beta-arrestin recruitment. The small constitutive activities of the mutants and the weak regulation of their functioning by agonist suggest a poor ability of the antidiuretic function to be adapted to the external stimuli, giving to the environmental factors an importance which can explain some of the phenotypic variability in patients having NSIAD. PMID- 20027298 TI - rBPI(21) promotes lipopolysaccharide aggregation and exerts its antimicrobial effects by (hemi)fusion of PG-containing membranes. AB - Antimicrobial peptides (AMPs) are important potential alternatives to conventional therapies against bacterial infections. rBPI(21) is a 21 kDa peptide based on the N-terminal region of the neutrophil bactericidal/permeability increasing protein (BPI). This AMP possesses highly selective bactericidal effects on Gram-negative bacteria and have affinity for lipopolysaccharide (LPS) which is believed to be at the origin of its neutralizing effect of the LPS segregated into the bloodstream. We aim at understanding the molecular bases of rBPI(21) bactericidal and LPS neutralization actions, using biomembrane model systems. Using dynamic light scattering spectroscopy we demonstrate that rBPI(21) promotes aggregation of negatively charged large unilamellar vesicles (LUV), even in the absence of LPS, and LPS aggregates, while for zwitterionic phosphatidylcholine (POPC) LUV the size remains unchanged. The peptide also promotes the fusion (or hemifusion) of membranes containing phosphatidylglycerol (POPG). The aggregation and fusion of negatively charged LUV are peptide concentration-dependent until massive aggregation is reached, followed by sample flocculation/precipitation. Concomitantly, there is a progressive change in the zeta-potential of the LUV systems and LPS aggregates. LUV systems composed of phosphatidylglycerol (POPG) and lipid mixtures with POPG have higher zeta potential variations than in the absence of POPG. The interaction of rBPI(21) with lipid vesicles is followed by leakage, with higher effect in POPG-containing membranes. LPS aggregation can be related with a decreased toxicity, possibly by facilitating its clearance by macrophage phagocytosis and/or blocking of LPS specific receptor recognition. Our data indicate that rBPI(21) mechanism of action at the molecular level involves the interaction with the LPS of the outer membrane of Gram-negative bacteria, followed by internalization and leakage induction through the (hemi)fusion of the bacterial outer and inner membranes, both enriched in phosphatidylglycerol. PMID- 20027299 TI - Transferability and fine-mapping of genome-wide associated loci for adult height across human populations. AB - Human height is the prototypical polygenic quantitative trait. Recently, several genetic variants influencing adult height were identified, primarily in individuals of East Asian (Chinese Han or Korean) or European ancestry. Here, we examined 152 genetic variants representing 107 independent loci previously associated with adult height for transferability in a well-powered sample of 1,016 unrelated African Americans. When we tested just the reported variants originally identified as associated with adult height in individuals of East Asian or European ancestry, only 8.3% of these loci transferred (p-values < or =0.05 under an additive genetic model with directionally consistent effects) to our African American sample. However, when we comprehensively evaluated all HapMap variants in linkage disequilibrium (r(2) > or = 0.3) with the reported variants, the transferability rate increased to 54.1%. The transferability rate was 70.8% for associations originally reported as genome-wide significant and 38.0% for associations originally reported as suggestive. An additional 23 loci were significantly associated but failed to transfer because of directionally inconsistent effects. Six loci were associated with adult height in all three groups. Using differences in linkage disequilibrium patterns between HapMap CEU or CHB reference data and our African American sample, we fine-mapped these six loci, improving both the localization and the annotation of these transferable associations. PMID- 20027300 TI - Loss-of-function mutations in Rab escort protein 1 (REP-1) affect intracellular transport in fibroblasts and monocytes of choroideremia patients. AB - BACKGROUND: Choroideremia (CHM) is a progressive X-linked retinopathy caused by mutations in the CHM gene, which encodes Rab escort protein-1 (REP-1), an escort protein involved in the prenylation of Rabs. Under-prenylation of certain Rabs, as a result of loss of function mutations in REP-1, could affect vesicular trafficking, exocytosis and secretion in peripheral cells of CHM patients. METHODOLOGY/PRINCIPAL FINDINGS: To evaluate this hypothesis, intracellular vesicle transport, lysosomal acidification and rates of proteolytic degradation were studied in monocytes (CD14+ fraction) and primary skin fibroblasts from the nine age-matched controls and thirteen CHM patients carrying 10 different loss-of function mutations. With the use of pHrodo BioParticles conjugated with E. coli, collagen I coated FluoSpheres beads and fluorescent DQ ovalbumin with BODYPY FL dye, we demonstrated for the first time that lysosomal pH was increased in monocytes of CHM patients and, as a consequence, the rates of proteolytic degradation were slowed. Microarray analysis of gene expression revealed that some genes involved in the immune response, small GTPase regulation, transcription, cell adhesion and the regulation of exocytosis were significantly up and down regulated in cells from CHM patients compared to controls. Finally, CHM fibroblasts secreted significantly lower levels of cytokine/growth factors such as macrophage chemoattractant protein-1 (MCP-1), pigment epithelial derived factor (PEDF), tumor necrosis factor (TNF) alpha, fibroblast growth factor (FGF) beta and interleukin (lL)-8. CONCLUSIONS/SIGNIFICANCE: We demonstrated for the first time that peripheral cells of CHM patients had increased pH levels in lysosomes, reduced rates of proteolytic degradation and altered secretion of cytokines. Peripheral cells from CHM patients expose characteristics that were not previously recognized and could used as an alternative models to study the effects of different mutations in the REP-1 gene on mechanism of CHM development in human population. PMID- 20027301 TI - Dissecting the role of critical residues and substrate preference of a Fatty Acyl CoA Synthetase (FadD13) of Mycobacterium tuberculosis. AB - Newly emerging multi-drug resistant strains of Mycobacterium tuberculosis (M.tb) severely limit the treatment options for tuberculosis (TB); hence, new antitubercular drugs are urgently needed. The mymA operon is essential for the virulence and intracellular survival of M.tb and thus represents an attractive target for the development of new antitubercular drugs. This study is focused on the structure-function relationship of Fatty Acyl-CoA Synthetase (FadD13, Rv3089) belonging to the mymA operon. Eight site-directed mutants of FadD13 were designed, constructed and analyzed for the structural-functional integrity of the enzyme. The study revealed that mutation of Lys(487) resulted in approximately 95% loss of the activity thus demonstrating its crucial requirement for the enzymatic activity. Comparison of the kinetic parameters showed the residues Lys(172) and Ala(302) to be involved in the binding of ATP and Ser(404) in the binding of CoenzymeA. The influence of mutations of the residues Val(209) and Trp(377) emphasized their importance in maintaining the structural integrity of FadD13. Besides, we show a synergistic influence of fatty acid and ATP binding on the conformation and rigidity of FadD13. FadD13 represents the first Fatty Acyl CoA Synthetase to display biphasic kinetics for fatty acids. FadD13 exhibits a distinct preference for C(26)/C(24) fatty acids, which in the light of earlier reported observations further substantiates the role of the mymA operon in remodeling the cell envelope of intracellular M.tb under acidic conditions. A three-dimensional model of FadD13 was generated; the docking of ATP to the active site verified its interaction with Lys(172), Ala(302) and Lys(487) and corresponded well with the results of the mutational studies. Our study provides a significant understanding of the FadD13 protein including the identification of residues important for its activity as well as in the maintenance of structural integrity. We believe that the findings of this study will provide valuable inputs in the development of inhibitors against the mymA operon, an important target for the development of antitubercular drugs. PMID- 20027302 TI - NPR1 protein regulates pathogenic and symbiotic interactions between Rhizobium and legumes and non-legumes. AB - BACKGROUND: Legumes are unique in their ability to establish symbiotic interaction with rhizobacteria from Rhizobium genus, which provide them with available nitrogen. Nodulation factors (NFs) produced by Rhizobium initiate legume root hair deformation and curling that entrap the bacteria, and allow it to grow inside the plant. In contrast, legumes and non-legumes activate defense responses when inoculated with pathogenic bacteria. One major defense pathway is mediated by salicylic acid (SA). SA is sensed and transduced to downstream defense components by a redox-regulated protein called NPR1. METHODOLOGY/PRINCIPAL FINDINGS: We used Arabidopsis mutants in SA defense pathway to test the role of NPR1 in symbiotic interactions. Inoculation of Sinorhizobium meliloti or purified NF on Medicago truncatula or nim1/npr1 A. thaliana mutants induced root hair deformation and transcription of early and late nodulins. Application of S. meliloti or NF on M. truncatula or A. thaliana roots also induced a strong oxidative burst that lasted much longer than in plants inoculated with pathogenic or mutualistic bacteria. Transient overexpression of NPR1 in M. truncatula suppressed root hair curling, while inhibition of NPR1 expression by RNAi accelerated curling. CONCLUSIONS/SIGNIFICANCE: We show that, while NPR1 has a positive effect on pathogen resistance, it has a negative effect on symbiotic interactions, by inhibiting root hair deformation and nodulin expression. Our results also show that basic plant responses to Rhizobium inoculation are conserved in legumes and non-legumes. PMID- 20027303 TI - Bone marrow osteoblastic niche: a new model to study physiological regulation of megakaryopoiesis. AB - BACKGROUND: The mechanism by which megakaryocytes (Mks) proliferate, differentiate, and release platelets into circulation are not well understood. Growing evidence indicates that a complex regulatory mechanism, involving cellular interactions, composition of the extracellular matrix and physical parameters such as oxygen tension, may contribute to the quiescent or permissive microenvironment related to Mk differentiation and maturation within the bone marrow. METHODOLOGY/PRINCIPAL FINDINGS: Differentiating human mesenchymal stem cells (hMSCs) into osteoblasts (hOSTs), we established an in vitro model for the osteoblastic niche. We demonstrated for the first time that the combination of HSCs, Mks and hypoxia sustain and promote bone formation by increasing type I collagen release from hOSTs and enhancing its fibrillar organization, as revealed by second harmonic generation microscopy. Through co-culture, we demonstrated that direct cell-cell contact modulates Mk maturation and differentiation. In particular we showed that low oxygen tension and direct interaction of hematopoietic stem cells (HSCs) with hOSTs inhibits Mk maturation and proplatelet formation (PPF). This regulatory mechanism was dependent on the fibrillar structure of type I collagen released by hOSTs and on the resulting engagement of the alpha2beta1 integrin. In contrast, normoxic conditions and the direct interaction of HSCs with undifferentiated hMSCs promoted Mk maturation and PPF, through a mechanism involving the VCAM-1 pathway. CONCLUSIONS/SIGNIFICANCE: By combining cellular, physical and biochemical parameters, we mimicked an in vitro model of the osteoblastic niche that provides a physiological quiescent microenvironment where Mk differentiation and PPF are prevented. These findings serve as an important step in developing suitable in vitro systems to use for the study and manipulation of Mk differentiation and maturation in both normal and diseased states. PMID- 20027304 TI - JNK1 phosphorylates SIRT1 and promotes its enzymatic activity. AB - SIRT1 is a NAD-dependent deacetylase that regulates a variety of pathways including the stress protection pathway. SIRT1 deacetylates a number of protein substrates, including histones, FOXOs, PGC-1alpha, and p53, leading to cellular protection. We identified a functional interaction between cJUN N-terminal kinase (JNK1) and SIRT1 by coimmunoprecipitation of endogenous proteins. The interaction between JNK1 and SIRT1 was identified under conditions of oxidative stress and required activation of JNK1 via phosphorylation. Modulation of SIRT1 activity or protein levels using nicotinamide or RNAi did not modify JNK1 activity as measured by its ability to phosphorylate cJUN. In contrast, human SIRT1 was phosphorylated by JNK1 on three sites: Ser27, Ser47, and Thr530 and this phosphorylation of SIRT1 increased its nuclear localization and enzymatic activity. Surprisingly, JNK1 phosphorylation of SIRT1 showed substrate specificity resulting in deacetylation of histone H3, but not p53. These findings identify a mechanism for regulation of SIRT1 enzymatic activity in response to oxidative stress and shed new light on its role in the stress protection pathway. PMID- 20027305 TI - Androgen regulated genes in human prostate xenografts in mice: relation to BPH and prostate cancer. AB - Benign prostatic hyperplasia (BPH) and prostate carcinoma (CaP) are linked to aging and the presence of androgens, suggesting that androgen regulated genes play a major role in these common diseases. Androgen regulation of prostate growth and development depends on the presence of intact epithelial-stromal interactions. Further, the prostatic stroma is implicated in BPH. This suggests that epithelial cell lines are inadequate to identify androgen regulated genes that could contribute to BPH and CaP and which could serve as potential clinical biomarkers. In this study, we used a human prostate xenograft model to define a profile of genes regulated in vivo by androgens, with an emphasis on identifying candidate biomarkers. Benign transition zone (TZ) human prostate tissue from radical prostatectomies was grafted to the sub-renal capsule site of intact or castrated male immunodeficient mice, followed by the removal or addition of androgens, respectively. Microarray analysis of RNA from these tissues was used to identify genes that were; 1) highly expressed in prostate, 2) had significant expression changes in response to androgens, and, 3) encode extracellular proteins. A total of 95 genes meeting these criteria were selected for analysis and validation of expression in patient prostate tissues using quantitative real time PCR. Expression levels of these genes were measured in pooled RNAs from human prostate tissues with varying severity of BPH pathologic changes and CaP of varying Gleason score. A number of androgen regulated genes were identified. Additionally, a subset of these genes were over-expressed in RNA from clinical BPH tissues, and the levels of many were found to correlate with disease status. Our results demonstrate the feasibility, and some of the problems, of using a mouse xenograft model to characterize the androgen regulated expression profiles of intact human prostate tissues. PMID- 20027306 TI - High prevalence of multidrug-tolerant bacteria and associated antimicrobial resistance genes isolated from ornamental fish and their carriage water. AB - BACKGROUND: Antimicrobials are used to directly control bacterial infections in pet (ornamental) fish and are routinely added to the water these fish are shipped in to suppress the growth of potential pathogens during transport. METHODOLOGY/PRINCIPAL FINDINGS: To assess the potential effects of this sustained selection pressure, 127 Aeromonas spp. isolated from warm and cold water ornamental fish species were screened for tolerance to 34 antimicrobials. Representative isolates were also examined for the presence of 54 resistance genes by a combination of miniaturized microarray and conventional PCR. Forty seven of 94 Aeromonas spp. isolates recovered from tropical ornamental fish and their carriage water were tolerant to > or =15 antibiotics, representing seven or more different classes of antimicrobial. The quinolone and fluoroquinolone resistance gene, qnrS2, was detected at high frequency (37% tested recent isolates were positive by PCR). Class 1 integrons, IncA/C broad host range plasmids and a range of other antibiotic resistance genes, including floR, bla(TEM-1), tet(A), tet(D), tet(E), qacE2, sul1, and a number of different dihydrofolate reductase and aminoglycoside transferase coding genes were also detected in carriage water samples and bacterial isolates. CONCLUSIONS: These data suggest that ornamental fish and their carriage water act as a reservoir for both multi-resistant bacteria and resistance genes. PMID- 20027307 TI - Expression of the VP2 protein of murine norovirus by a translation termination reinitiation strategy. AB - BACKGROUND: Expression of the minor virion structural protein VP2 of the calicivirus murine norovirus (MNV) is believed to occur by the unusual mechanism of termination codon-dependent reinitiation of translation. In this process, following translation of an upstream open reading frame (ORF) and termination at the stop codon, a proportion of 40S subunits remain associated with the mRNA and reinitiate at the AUG of a downstream ORF, which is typically in close proximity. Consistent with this, the VP2 start codon (AUG) of MNV overlaps the stop codon of the upstream VP1 ORF (UAA) in the pentanucleotide UAAUG. PRINCIPAL FINDINGS: Here, we confirm that MNV VP2 expression is regulated by termination-reinitiation and define the mRNA sequence requirements. Efficient reintiation is dependent upon 43 nt of RNA immediately upstream of the UAAUG site. Chemical and enzymatic probing revealed that the RNA in this region is not highly structured and includes an essential stretch of bases complementary to 18S rRNA helix 26 (Motif 1). The relative position of Motif 1 with respect to the UAAUG site impacts upon the efficiency of the process. Termination-reinitiation in MNV was also found to be relatively insensitive to the initiation inhibitor edeine. CONCLUSIONS: The termination-reinitiation signal of MNV most closely resembles that of influenza BM2. Similar to other viruses that use this strategy, base-pairing between mRNA and rRNA is likely to play a role in tethering the 40S subunit to the mRNA following termination at the VP1 stop codon. Our data also indicate that accurate recognition of the VP2 ORF AUG is not a pre-requisite for efficient reinitiation of translation in this system. PMID- 20027308 TI - Development of functional human NK cells in an immunodeficient mouse model with the ability to provide protection against tumor challenge. AB - Studies of human NK cells and their role in tumor suppression have largely been restricted to in vitro experiments which lack the complexity of whole organisms, or mouse models which differ significantly from humans. In this study we showed that, in contrast to C57BL/6 Rag2(-/-)/gamma(c) (-/-) and NOD/Scid mice, newborn BALB/c Rag2(-/-)/gamma(c) (-/-) mice can support the development of human NK cells and CD56+ T cells after intrahepatic injection with hematopoietic stem cells. The human CD56(+) cells in BALB/c Rag2(-/-)/gamma(c) (-/-) mice were able to produce IFN-gamma in response to human IL-15 and polyI:C. NK cells from reconstituted Rag2(-/-)/gamma(c) (-/-) mice were also able to kill and inhibit the growth of K562 cells in vitro and were able to produce IFN-gamma in response to stimulation with K562 cells. In vivo, reconstituted Rag2(-/-)/gamma(c) (-/-) mice had higher survival rates after K562 challenge compared to non-reconstituted Rag2(-/-)/gamma(c) (-/-) mice and were able to control tumor burden in various organs. Reconstituted Rag2(-/-)/gamma(c) (-/-) mice represent a model in which functional human NK and CD56+ T cells can develop from stem cells and can thus be used to study human disease in a more clinically relevant environment. PMID- 20027309 TI - Nuclear receptor unfulfilled regulates axonal guidance and cell identity of Drosophila mushroom body neurons. AB - Nuclear receptors (NRs) comprise a family of ligand-regulated transcription factors that control diverse critical biological processes including various aspects of brain development. Eighteen NR genes exist in the Drosophila genome. To explore their roles in brain development, we knocked down individual NRs through the development of the mushroom bodies (MBs) by targeted RNAi. Besides recapitulating the known MB phenotypes for three NRs, we found that unfulfilled (unf), an ortholog of human photoreceptor specific nuclear receptor (PNR), regulates axonal morphogenesis and neuronal subtype identity. The adult MBs develop through remodeling of gamma neurons plus de-novo elaboration of both alpha'/beta' and alpha/beta neurons. Notably, unf is largely dispensable for the initial elaboration of gamma neurons, but plays an essential role in their re extension of axons after pruning during early metamorphosis. The subsequently derived MB neuron types also require unf for extension of axons beyond the terminus of the pruned bundle. Tracing single axons revealed misrouting rather than simple truncation. Further, silencing unf in single-cell clones elicited misguidance of axons in otherwise unperturbed MBs. Such axon guidance defects may occur as MB neurons partially lose their subtype identity, as evidenced by suppression of various MB subtype markers in unf knockdown MBs. In sum, unf governs axonal morphogenesis of multiple MB neuron types, possibly through regulating neuronal subtype identity. PMID- 20027310 TI - A real-time PCR array for hierarchical identification of Francisella isolates. AB - A robust, rapid and flexible real-time PCR assay for hierarchical genetic typing of clinical and environmental isolates of Francisella is presented. Typing markers were found by multiple genome and gene comparisons, from which 23 canonical single nucleotide polymorphisms (canSNPs) and 11 canonical insertion deletion mutations (canINDELs) were selected to provide phylogenetic guidelines for classification from genus to isolate level. The specificity of the developed assay, which uses 68 wells of a 96-well real-time PCR format with a detection limit of 100 pg DNA, was assessed using 62 Francisella isolates of diverse genetic and geographical origins. It was then successfully used for typing 14 F. tularensis subsp. holarctica isolates obtained from tularemia patients in Sweden in 2008 and five more genetically diverse Francisella isolates of global origins. When applied to human ulcer specimens for direct pathogen detection the results were incomplete due to scarcity of DNA, but sufficient markers were identified to detect fine-resolution differences among F. tularensis subsp. holarctica isolates causing infection in the patients. In contrast to other real-time PCR assays for Francisella, which are typically designed for specific detection of a species, subspecies, or strain, this type of assay can be easily tailored to provide appropriate phylogenetic and/or geographical resolution to meet the objectives of the analysis. PMID- 20027311 TI - Pebble and rock band: heuristic resolution of repeats and scaffolding in the velvet short-read de novo assembler. AB - BACKGROUND: Despite the short length of their reads, micro-read sequencing technologies have shown their usefulness for de novo sequencing. However, especially in eukaryotic genomes, complex repeat patterns are an obstacle to large assemblies. PRINCIPAL FINDINGS: We present a novel heuristic algorithm, Pebble, which uses paired-end read information to resolve repeats and scaffold contigs to produce large-scale assemblies. In simulations, we can achieve weighted median scaffold lengths (N50) of above 1 Mbp in Bacteria and above 100 kbp in more complex organisms. Using real datasets we obtained a 96 kbp N50 in Pseudomonas syringae and a unique 147 kbp scaffold of a ferret BAC clone. We also present an efficient algorithm called Rock Band for the resolution of repeats in the case of mixed length assemblies, where different sequencing platforms are combined to obtain a cost-effective assembly. CONCLUSIONS: These algorithms extend the utility of short read only assemblies into large complex genomes. They have been implemented and made available within the open-source Velvet short-read de novo assembler. PMID- 20027312 TI - Crawling to collapse: ecologically unsound ornamental invertebrate fisheries. AB - BACKGROUND: Fishery management has historically been an inexact and reactionary discipline, often taking action only after a critical stock suffers overfishing or collapse. The invertebrate ornamental fishery in the State of Florida, with increasing catches over a more diverse array of species, is poised for collapse. Current management is static and the lack of an adaptive strategy will not allow for adequate responses associated with managing this multi-species fishery. The last decade has seen aquarium hobbyists shift their display preference from fish only tanks to miniature reef ecosystems that include many invertebrate species, creating increased demand without proper oversight. The once small ornamental fishery has become an invertebrate-dominated major industry supplying five continents. METHODOLOGY/PRINCIPAL FINDINGS: Here, we analyzed the Florida Marine Life Fishery (FLML) landing data from 1994 to 2007 for all invertebrate species. The data were organized to reflect both ecosystem purpose (in the wild) and ecosystem services (commodities) for each reported species to address the following question: Are ornamental invertebrates being exploited for their fundamental ecosystem services and economic value at the expense of reef resilience? We found that 9 million individuals were collected in 2007, 6 million of which were grazers. CONCLUSIONS/SIGNIFICANCE: The number of grazers now exceeds, by two-fold, the number of specimens collected for curio and ornamental purposes altogether, representing a major categorical shift. In general, landings have increased 10-fold since 1994, though the number of licenses has been dramatically reduced. Thus, despite current management strategies, the FLML Fishery appears to be crawling to collapse. PMID- 20027313 TI - Multiple lineages of human breast cancer stem/progenitor cells identified by profiling with stem cell markers. AB - Heterogeneity of cancer stem/progenitor cells that give rise to different forms of cancer has been well demonstrated for leukemia. However, this fundamental concept has yet to be established for solid tumors including breast cancer. In this communication, we analyzed solid tumor cancer stem cell markers in human breast cancer cell lines and primary specimens using flow cytometry. The stem/progenitor cell properties of different marker expressing-cell populations were further assessed by in vitro soft agar colony formation assay and the ability to form tumors in NOD/SCID mice. We found that the expression of stem cell markers varied greatly among breast cancer cell lines. In MDA-MB-231 cells, PROCR and ESA, instead of the widely used breast cancer stem cell markers CD44(+)/CD24(-/low) and ALDH, could be used to highly enrich cancer stem/progenitor cell populations which exhibited the ability to self renew and divide asymmetrically. Furthermore, the PROCR(+)/ESA(+) cells expressed epithelial-mesenchymal transition markers. PROCR could also be used to enrich cells with colony forming ability from MB-361 cells. Moreover, consistent with the marker profiling using cell lines, the expression of stem cell markers differed greatly among primary tumors. There was an association between metastasis status and a high prevalence of certain markers including CD44(+)/CD24(-/low), ESA(+), CD133(+), CXCR4(+) and PROCR(+) in primary tumor cells. Taken together, these results suggest that similar to leukemia, several stem/progenitor cell-like subpopulations can exist in breast cancer. PMID- 20027314 TI - Substantial contribution of submicroscopical Plasmodium falciparum gametocyte carriage to the infectious reservoir in an area of seasonal transmission. AB - BACKGROUND: Man to mosquito transmission of malaria depends on the presence of the sexual stage parasites, gametocytes, that often circulate at low densities. Gametocyte densities below the microscopical threshold of detection may be sufficient to infect mosquitoes but the importance of submicroscopical gametocyte carriage in different transmission settings is unknown. METHODOLOGY/PRINCIPAL FINDINGS: Membrane feeding experiments were carried out on 80 children below 14 years of age at the end of the wet season in an area of seasonal malaria transmission in Burkina Faso. Gametocytes were quantified by microscopy and by Pfs25-based quantitative nucleic acid sequence-based amplification assay (QT NASBA). The children's infectiousness was determined by membrane feeding experiments in which a venous blood sample was offered to locally reared Anopheles mosquitoes. Gametocytes were detected in 30.0% (24/80) of the children by microscopy compared to 91.6% (65/71) by QT-NASBA (p<0.001). We observed a strong association between QT-NASBA gametocyte density and infection rates (p = 0.007). Children with microscopically detectable gametocytes were more likely to be infectious (68.2% compared to 31.7% of carriers of submicroscopical gametocytes, p = 0.001), and on average infected more mosquitoes (13.2% compared to 2.3%, p<0.001). However, because of the high prevalence of submicroscopical gametocyte carriage in the study population, carriers of sub-microscopical gametocytes were responsible for 24.2% of the malaria transmission in this population. CONCLUSIONS/SIGNIFICANCE: Submicroscopical gametocyte carriage is common in an area of seasonal transmission in Burkina Faso and contributes substantially to the human infectious reservoir. Submicroscopical gametocyte carriage should therefore be considered when implementing interventions that aim to reduce malaria transmission. PMID- 20027315 TI - An adequately robust early TNF-alpha response is a hallmark of survival following trauma/hemorrhage. AB - BACKGROUND: Trauma/hemorrhagic shock (T/HS) results in cytokine-mediated acute inflammation that is generally considered detrimental. METHODOLOGY/PRINCIPAL FINDINGS: Paradoxically, plasma levels of the early inflammatory cytokine TNF alpha (but not IL-6, IL-10, or NO(2) (-)/NO(3) (-)) were significantly elevated within 6 h post-admission in 19 human trauma survivors vs. 4 non-survivors. Moreover, plasma TNF-alpha was inversely correlated with Marshall Score, an index of organ dysfunction, both in the 23 patients taken together and in the survivor cohort. Accordingly, we hypothesized that if an early, robust pro-inflammatory response were to be a marker of an appropriate response to injury, then individuals exhibiting such a response would be predisposed to survive. We tested this hypothesis in swine subjected to various experimental paradigms of T/HS. Twenty-three anesthetized pigs were subjected to T/HS (12 HS-only and 11 HS + Thoracotomy; mean arterial pressure of 30 mmHg for 45-90 min) along with surgery only controls. Plasma obtained at pre-surgery, baseline post-surgery, beginning of HS, and every 15 min thereafter until 75 min (in the HS only group) or 90 min (in the HS + Thoracotomy group) was assayed for TNF-alpha, IL-6, IL-10, and NO(2) (-)/NO(3) (-). Mean post-surgery+/-HS TNF-alpha levels were significantly higher in the survivors vs. non-survivors, while non-survivors exhibited no measurable change in TNF-alpha levels over the same interval. CONCLUSIONS/SIGNIFICANCE: Contrary to the current dogma, survival in the setting of severe, acute T/HS appears to be associated with an immediate increase in serum TNF-alpha. It is currently unclear if this response was the cause of this protection, a marker of survival, or both. This abstract won a Young Investigator Travel Award at the SHOCK 2008 meeting in Cologne, Germany. PMID- 20027316 TI - Expression profiling the temperature-dependent amphibian response to infection by Batrachochytrium dendrobatidis. AB - Amphibians are experiencing a panzootic of unprecedented proportions caused by the emergence of Batrachochytrium dendrobatidis (Bd). However, all species are not equally at risk of infection, and risk is further modified by environmental variables, specifically temperature. In order to understand how, and when, hosts mount a response to Bd we analysed infection dynamics and patterns of gene expression in the model amphibian species Silurana (Xenopus) tropicalis. Mathematical modelling of infection dynamics demonstrate the existence of a temperature-dependent protective response that is largely independent of the intrinsic growth-rate of Bd. Using temporal expression-profiling by microarrays and qRT-PCR, we characterise this response in the main amphibian lymphoid tissue, the spleen. We demonstrate that clearance of Bd at the host-optimal temperature is not clearly associated with an adaptive immune response, but rather is correlated with the induction of components of host innate immunity including the expression of genes that are associated with the production of the antimicrobial skin peptide preprocareulein (PPCP) as well as inflammatory responses. We find that adaptive immunity appears to be lacking at host-optimal temperatures. This suggests that either Bd does not stimulate, or suppresses, adaptive immunity, or that trade-offs exist between innate and adaptive limbs of the amphibian immune system. At cold temperatures, S. tropicalis loses the ability to mount a PPCP based innate response, and instead manifests a more pronounced inflammatory reaction that is characterised by the production of proteases and higher pathogen burdens. This study demonstrates the temperature-dependency of the amphibian response to infection by Bd and indicates the influence that changing climates may exert on the ectothermic host response to pathogens. PMID- 20027318 TI - Facilitation of acetylcholine secretion in mouse motor synapses caused by calcium release from depots upon activation of L-type calcium channels. AB - Pharmacological disinhibition of L-type Ca(2+) channels by two ways (with agonist S(-) BAY K 8644 and iberiotoxin, a Ca(2+)-activated BK-type K(+)-channel blocker) increases quantal content of evoked end-plate potentials, which was completely prevented by ryanodine (2 microM) blockade of ryanodine receptors. We conclude that increased quantal secretion of the transmitter induced by L-type Ca(2+) channel functioning requires activation of ryanodine receptors and calcium release from depots in motor terminals in mice. PMID- 20027317 TI - Flotillins are involved in the polarization of primitive and mature hematopoietic cells. AB - BACKGROUND: Migration of mature and immature leukocytes in response to chemokines is not only essential during inflammation and host defense, but also during development of the hematopoietic system. Many molecules implicated in migratory polarity show uniform cellular distribution under non-activated conditions, but acquire a polarized localization upon exposure to migratory cues. METHODOLOGY/PRINCIPAL FINDINGS: Here, we present evidence that raft-associated endocytic proteins (flotillins) are pre-assembled in lymphoid, myeloid and primitive hematopoietic cells and accumulate in the uropod during migration. Furthermore, flotillins display a polarized distribution during immunological synapse formation. Employing the membrane lipid-order sensitive probe Laurdan, we show that flotillin accumulation in the immunological synapse is concomittant with membrane ordering in these regions. CONCLUSIONS: Together with the observation that flotillin polarization does not occur in other polarized cell types such as polarized epithelial cells, our results suggest a specific role for flotillins in hematopoietic cell polarization. Based on our results, we propose that in hematopoietic cells, flotillins provide intrinsic cues that govern segregation of certain microdomain-associated molecules during immune cell polarization. PMID- 20027319 TI - Role of neuronal NMDA and non-NMDA glutamate receptors in medial vestibular nucleus in the regulation of respiratory rhythmogenesis in newborn rats in vitro. AB - We studied isolated pontobulbospinal preparations from newborn rat brain. In the early postnatal period, the rostral part of the medial vestibular nucleus produces a potent inhibitory effect on neuronal structures of the bulbar respiratory center via the glutamatergic system. Microinjection of L-glutamate (50 mmol/liter) into the rostral part of the vestibular nucleus completely blocks respiratory rhythmogenesis in 0-1-day-old rat pups and reduced the frequency of generation of inspiratory discharges in 2-3-day-old rats from 8.42+/-0.68 to 2.68+/-0.32 min(-1). It was found that the leading role in the mechanism of glutamatergic modulation of the respiratory rhythmogenesis by neurons of the medial vestibular nucleus is played by NMDA and, to a lesser extent, non-NMDA glutamate receptors. PMID- 20027320 TI - Effect of calcium-regulating hormones and calcium channel modulators on glucose consumption by muscle and adipose tissues in vivo and in vitro. AB - We studied the effects of calcitonin, parathyrin, and Ca(2+) channel antagonist isoptin and agonist Bay-K-8644 on glucose consumption by muscle (diaphragm) and adipose (epididymal) tissues and insulin-stimulated glucose consumption in vivo and in vitro. Calcitonin and parathyrin did not alter glucose consumption; parathyrin did not affect, while calcitonin completely abolished the stimulating effect of insulin in vivo and in vitro. Isoptin significantly increased glucose consumption in vivo and in vitro, while Bay-K-8644 in vitro had no effect glucose consumption. Isoptin did not affect, while Bay-K-8644 significantly reduced the stimulating effect of insulin on glucose consumption by the muscle and adipose tissues. Isoptin did not affect the stimulating effect of insulin against the background of parathyrin administration and completely blocked the inhibitory effect of calcitonin on insulin-stimulated glucose consumption by the muscle and adipose tissues in vivo and in vitro, while Bay-K-8644 potentiated this effect of calcitonin in vitro. PMID- 20027322 TI - Rhythmoinotropic myocardial reactions in rats with postinfarction cardiosclerosis against the background of streptozotocin-induced diabetes. AB - We studied rhythmoinotropic reactions of the myocardium in rats with postinfarction cardiosclerosis and in rats with postinfarction cardiosclerosis against the background of streptozotocin-induced diabetes. Inotropic myocardial response in rats with postinfarction cardiosclerosis was significantly inhibited after rest periods, while in streptozotocin diabetic rats the rhythmoinotropic myocardial reaction was comparable with the reaction of intact myocardium. The combination of postinfarction cardiosclerosis and diabetes paradoxically contributed to preservation of contractile function of the myocardium in rats. PMID- 20027321 TI - Allelic polymorphism of cytokine genes during pulmonary tuberculosis. AB - Modern immunological and molecular genetic studies showed that tuberculosis is accompanied by an imbalance in the production of immunoregulatory cytokines by mononuclear leukocytes. T allele and homozygous TT genotype of T-330G polymorphism in the IL2 gene, T allele and TT genotype of C-590T polymorphism in the IL4 gene, and CC genotype of A-1188C polymorphism in the IL12B gene are immunogenetic factors that have protective activity against susceptibility to pulmonary tuberculosis. Susceptibility to tuberculous infection is associated with A1A2 genotype of the polymorphic region +3953 A1/A2 in the IL1B gene; G allele and TG and GG genotypes of T-330G polymorphism in the IL2 gene; C allele and CC and CT genotypes of C-590T polymorphism in the IL4 gene; and AC genotype of the polymorphic region A-1188C in the IL12 gene. PMID- 20027323 TI - Neurotransmitter mechanisms of morphine withdrawal syndrome. AB - Activity of the major neurotransmitter systems in the brainstem and cerebellum was studied in rats with morphine withdrawal syndrome. The most significant changes were found in the brainstem of animals by the 36th hour and 7 days after morphine withdrawal. Dysfunction was revealed in the dopaminergic, noradrenergic, and serotoninergic neurotransmitter systems. Changes in cerebellar neurotransmission were most pronounced by the end of the first week of morphine withdrawal syndrome and manifested in the prevalence of inhibitory processes. PMID- 20027324 TI - Effect of ceruloplasmin on the number and resistance of erythrocytes during acute physical exercise. AB - Acute physical exercise was followed by a decrease in the osmotic resistance of erythrocytes, shortening of the time-to-onset of erythrocyte hemolysis, and increase in the sorption capacity of the cell membrane. Administration of ceruloplasmin 24 h before physical exercise normalizes membrane resistance in red blood cells. PMID- 20027325 TI - Kinetic parameters of liver aldehyde dehydrogenase in rats with cold injury. AB - We studied the kinetics of liver aldehyde dehydrogenase in rats with cold injury. Low-temperature exposure was followed by a decrease in activity and catalytic efficiency of aldehyde dehydrogenase in rat liver mitochondria. Atypical changes in kinetic characteristics of aldehyde dehydrogenase were found in the cytoplasmic fraction during cold injury. PMID- 20027326 TI - Nitroxidergic neurons in nuclei of the medulla oblongata in hypertensive and normotensive rats. AB - Distribution of nitroxidergic neurons and neuronal NO-synthase activity in certain medulla oblongata nuclei were studied in normotensive and hypertensive rats with different types of arterial hypertension. In rats with renovascular hypertension, neuronal NO-synthase activity markedly decreased in most nuclei 2 weeks after surgery, while the number of NO-positive cells did not change significantly; after 4 weeks, the percentage of NO-positive neuron markedly decreased and neuronal NO-synthase activity also slightly decreased. No further decrease in neuronal NO-synthase activity was observed 8 week after intervention, but the percentage of NO-neurons decreased compared to that in normotensive rats. In spontaneously hypertensive rats, changes in most nuclei were directed similarly to those observed in 8-week renovascular hypertension, but motor nuclei demonstrated less differentiated reaction to hypertension. In all cases, changes in the parameters observed in sensory nucleus (n. solitarius) appeared earlier and were more pronounced than in nuclei of the reticular formation related to bulbar vasomotor center. PMID- 20027327 TI - Blood cytokines in rats with various behavioral characteristics during emotional stress and treatment with interleukin-1beta. AB - We studied the effect of acute emotional stress and exogenous IL-1beta (5 microg/kg intraperitoneally) on the cytokine profile of blood serum in Wistar rats with various behavioral characteristics in the open-field test. Blood level of proinflammatory cytokine IL-1beta decreased in behaviorally passive rats, but increased in active animals after simultaneous immobilization and electrocutaneous stimulation. These changes reflect the opposite immune responses to a similar stress exposure in rats with different emotional reactivity. Poststress variations in the concentration of circulating IL-1beta differed in rats receiving exogenous IL-1beta. Blood cytokine concentration decreased in behaviorally active rats, but remained unchanged in passive animals that were exposed to immobilization and electrocutaneous stimulation after pretreatment with IL-1beta. Emotional stress and injection of IL-1beta had no effect on blood level of an anti-inflammatory cytokine IL-4 in rats. Our results indicate that rats with various behavioral parameters are characterized by significant differences in the cytokine profile of blood serum under conditions of emotional stress and treatment with IL-1beta. These data illustrate the specific functional features of immune mechanisms, which provide an individual resistance of rats to the same stress exposure. PMID- 20027328 TI - Effects of terahertz irradiation at nitric oxide frequencies on intensity of lipoperoxidation and antioxidant properties of the blood under stress conditions. AB - The effects of terahertz irradiation at the nitric oxide frequencies (150.176 150.664 GHz) on the intensity of lipoperoxidation and antioxidant properties of the blood were examined on albino rats subjected to immobilization stress. This terahertz irradiation completely normalized LPO processes and functional activity of antioxidants in stressed rats. PMID- 20027329 TI - Phagocytic activity of alveolar macrophages in the presence of liposomal "biene". AB - Phagocytic activity of alveolar macrophages isolated from rat bronchoalveolar lavage fluid was studied during their reaction with multilamellar liposomes containing biene (a complex of unsaturated fatty acids). Liposomes with incorporated biene significantly stimulated phagocytic activity of freshly isolated macrophages and primary culture of these cells (judging from the increase in phagocytic index and phagocytic number). A quantitative relationship between the phagocytic number and phagocytic index, on the one hand, and the concentration of liposomal biene, on the other, was detected. The effects of liposomes containing incorporated biene, liposomes of similar composition without biene, and pure biene substance on cell suspension were compared. PMID- 20027330 TI - Effect of apolipoprotein a-I complex with tetrahydrocortisone on protein biosynthesis and glucose absorption by rat hepatocytes. AB - We studied the effect of apolipoprotein A-I-tetrahydrocortisone complex on (14)C glucose absorption and lactate accumulation and on the rate of protein biosynthesis in isolated rat hepatocytes. The presence of apolipoprotein A-I tetrahydrocortisone complex in the incubation medium increased absorption of labeled glucose by hepatocytes by 52%, while lactate content in the conditioning medium increased 4-fold. The rate of protein biosynthesis increased by 80% in comparison with control cells. It is hypothesized that the increase in protein biosynthesis rate in hepatocytes under the effect of apolipoprotein A-I tetrahydrocortisone complex is due to stimulation of energy metabolism, specifically, of its glycolytic component. PMID- 20027331 TI - ATP as modulator of carbacholine effect on contractility of rat myocardium in postnatal ontogeny. AB - We studied combined effect of 2-m-ATP, P(2) receptor agonist, and carbacholine, muscarinic M(2) cholinoreceptor agonist, on contractility of rat myocardium during the postnatal ontogeny. Activation of P(2) receptors can stimulate or attenuate the effects of carbacholine depending on animal age. 2-m-ATP potentiates the inhibitory effect of carbacholine on myocardial contractility in 14- and 100-day-old rats. In 21-day-old rats, activation of P(2) receptors prevented the negative effect of carbacholine on myocardial contractility. Activation of muscarinic M(2) receptors inhibited the inotropic effect of purine in all age groups. PMID- 20027332 TI - Study of antimycotic activity of lyticase. AB - Laboratory studies of lyticase (enzymatic drug) as an antimycotic agent were carried out. The enzyme reduced optical density of Candida albicans test culture, inhibited adhesion of yeast-like fungi on vaginal epitheliocytes, stimulated the formation of germinative tubes, and made Candida albicans more available for phagocytosis. PMID- 20027333 TI - Changes in copper metabolism in different compartments of the brain in rats with induced fibrillogenesis. AB - Fibrillogenesis was induced in rats by injection of a fragment of neurotoxic protein, beta-amyloid protein precursor, into the cerebral ventricle. Copper, iron, and zinc concentrations and relative activities of genes of copper transporting protein and extracellular and intracellular cuproenzymes were evaluated in different brain compartments of these animals. Copper and zinc concentrations decreased significantly in different compartments of the brain of rats with experimental fibrillogenesis, while iron content did not change. According to the data of RT-PCR analysis, activities of genes of copper transporting protein and extracellular coenzyme decreased. The expression of intracellular cuproenzyme genes and the content of SOD1 protein did not change, SOD1 activity in the cytosol decreased, and active SOD1 was detected in the mitochondrial intermembrane space. The relationship between fibrillogenesis and copper metabolism is discussed. PMID- 20027334 TI - Norepinephrine content in the hypothalamus and medulla oblongata of ISIAH rats is regulated by several genetic loci. AB - Five genetic loci regulating epinephrine content in rat hypothalamus and two loci in the medulla oblongata were identified using polymorphic microsatellite DNA markers. Allele polymorphism of these loci determines the differences between norepinephrine levels in brain compartments of hypertensive ISIAH and normotensive WAG rats. The interactions between the detected epinephrine regulating loci are additive. PMID- 20027335 TI - Expression of S100B and S100A6 genes during long-term posttetanic potentiation in the hippocampus. AB - The expression of S100B and S100A6 mRNA in CA1 region of rat hippocampal sections was studied after tetanizing stimulation. The level of S100B expression increased 2-4-fold in comparison with the control after 30 min and gradually returned to the basal level 120 min after tetanization. The level of S100A6 mRNA was very low and did not change after tetanization. PMID- 20027336 TI - Molecular markers of tumors. AB - The main biologically significant molecular markers of human tumors are discussed on the basis of modern published data and author's findings of many-year studies: steroid hormone receptors, growth factors and underlying signal proteins, tumor associated proteases, and angiogenesis markers. Methodological aspects, progress in preclinical studies, international recommendations on the use of these parameters for prediction of the disease course and prescription of effective therapy are analyzed. Latest data on the potentialities and limitations of modern highly productive technologies (microchips) as an alternative to studies of individual molecular markers are presented. PMID- 20027337 TI - A method for improving the efficiency of therapy for melanoma. AB - A method for photodynamic therapy was developed in experiments on the model of melanoma B16. The photosensitizer is injected by two doses, while laser exposure is carried out during coincidence of its maximum accumulation phases in tumor vessels and cells. This method increased the percentage of animals with complete regression of the tumor, decreased the coefficient of absolute tumor increase in animals with progressive tumor growth, increased survival of mice, and significantly decreased melanoma metastasizing to the lungs in comparison with untreated animals and even with the standard photodynamic therapy. PMID- 20027338 TI - Clinical significance of hyperexpression of epidermal growth factor receptors (EGFR and HER-2) in esophageal squamous cell carcinoma. AB - Immunohistochemical study of marker expression in primary tumors of patients with esophageal squamous-cell carcinoma was carried out in order to evaluate prognostic significance of EGFR and HER-2 levels in the tumors. Hyperexpression of EGFR and HER-2 in the tumors is an important marker for the analysis of prognosis and clinical course of the disease. A relationship between high levels of EGFR and HER-2 in the tumors of patients with esophageal squamous-cell carcinoma and intravascular tumor invasion (p=0.038) and poor outcome of the disease (p=0.019) was detected. The results indicate that evaluation of changes in the expression of EGFR and HER-2 in tumors is essential for individual prediction of the disease course and development of new approaches to the treatment of these tumors, including target therapy aimed at these tyrosine kinase receptors. PMID- 20027339 TI - Endostatin, placental growth factor, and fibroblast growth factors-1 and -2 in the sera of patients with primary osteosarcomas. AB - Serum levels of endostatin, placental growth factor (PlGF), and fibroblast growth factors-1 and -2 (FGF-1 and FGF-2) were measured in 58 patients with primary osteosarcomas before therapy and in 21 healthy subjects. The incidence of serum FGF-1 in bone tumors was 2.5 times higher than in healthy individuals (p=0.004); significant levels of FGF-2, PlGF, and endostatin were detected in all examined subjects. The mean serum level of endostatin in healthy individuals was significantly lower than in the total group of patients with bone tumors (p=0.005). The level of FGF-1 in osteosarcomas was significantly higher than in chondrosarcomas (p<0.05). No appreciable differences in FGF-2 levels were detected in patients with tumors of different histological structure. The mean serum content of PlGF was virtually the same in healthy individuals and patients with bone tumors. A significant relationship between serum PlGF level and maximum tumor size (p=0.008) was detected in osteosarcoma. No relationships between the levels of FGF-1, FGF-2, PlGF, and endostatin were detected in healthy subjects and patients with primary tumors of the bones. Differences in 3-year overall survival values of patients with bone sarcomas with different initial serum levels of FGF-1 and endostatin were detected. PMID- 20027340 TI - Parameters of the cytokine system functioning in laboratory primates. AB - The cytokine status (IFN, IL, etc.) of different monkey species (M. mulatta, P. hamadryas, C. aethiops) was studied. The interferon status is determined by the following parameters: IFN content in circulating blood and production of IFN alpha and IFN-gamma by lymphocytes after appropriate in vitro induction. The interferon status of monkeys is similar to that of humans. The capacity to produce IFN reduces with age. It was found that genes of virtually all studied cytokines are expressed in blood cells and hence, in immune system cells. PMID- 20027341 TI - Proliferation and survival of rat C6 glioma culture in the presence of implants coated with modified carbon-based films. AB - The survival of rat C6 glioma decreased in the presence of implants from VT-16 titanium alloy. Diamond-like carbon coating of VT-16 alloy slightly increased cell death on day 5 of the experiment (39.9+/-2.1%). The percentage of dead C6 glioma cells inside titanium rings with diamond-like carbon coating, incorporating up to 3.5 atom.% Ag nanoparticles, was 53.7+/-4.3% on day 5 of culturing, while after doping to 6.7 atom.% Ag cell death reached 66.7+/-3.2% (p<0.05). The maximum toxic effect towards C6 glioma was detected in the specimens coated with diamond-like film with silver nanoparticles. PMID- 20027342 TI - Morphological assessment of different ways of bronchial stump closure after pneumonectomy. AB - A method is developed to close the stump of the primary bronchus by outer compression with a shape-memory device. The process of bronchial stump healing under compression suture was morphologically examined. The novel method was compared with the routine Sweet-type hand-operated and the mechanical sutures. The compression suture heals the stump by primary intention with restoration of the typical airway epithelium on the stump bottom and prevention of the development of the bronchial fistulas. PMID- 20027343 TI - Association of brain weight value with morphometric and histochemical properties of cortical neurons and higher nervous activity parameters in rats in prepubertal period. AB - We compared thickness of the neocortex, morphometric and histochemical characteristics of neurons in frontoparietal and parietal lobes and hippocampal field I in animals from great litters with lower brain weight (group 1) and from small (artificially reduced) litters with higher brain weight (group 2). It was found that the thickness of the neocortex in the frontoparietal and parietal lobes does not differ in the compared groups, while the size of neuronal cytoplasm in layer II of the frontoparietal and parietal lobes and in layer V of the frontoparietal lobe in group 2 animals was lower than in group 1. Nuclei of cortical neurons in layer II of the parietal and frontoparietal lobes and in frontoparietal lobe layer V in group 2 animals were smaller than in group 1. Neuronal nucleoli in group 2 animals were also smaller than in group 1 rats. RNA concentration in neuronal cytoplasm in the hippocampus and neocortex of group 2 rats was higher than in group 1 animals. NADPH-dehydrogenase activity in neurons of parietal lobe (layer II) and hippocampus in group 2 rats was lower than in group 1 animals, NADH-dehydrogenase activity was lower in parietal lobe layer II neurons. Group 2 rats demonstrated increased number of hanging down, sniffing, movements, entries into open and closed arms, and lower immobility time in the elevated plus-maze test. PMID- 20027344 TI - The use of anaferon (pediatric formulation) for prophylaxis of acute respiratory viral infections in preschool children. AB - Anaferon (pediatric formulation) reduces the incidence of acute respiratory viral infections in sickly children and children with bronchial asthma and has a positive effect on the course of asthma. The preparation produces an immunomodulating effect (increases initially low IFN-gamma levels and normalizes elevated levels of IL-1beta), stimulates synthesis of IgA and IgG, exhibits cytoprotective activity, and improves local immunity of the upper airways in sickly children. PMID- 20027345 TI - Efficiency of interferon inductor anaferon (pediatric formulation) in prophylaxis of acute respiratory infections in sickly children. AB - Prophylactic efficiency and safety of anaferon (pediatric formulation) in children aging 1?month to 4 years, including sickly children, was proven. The use of the preparation in children reduced the incidence of acute respiratory infections, alleviated the course of the disease, and decreased the incidence of detection of viral antigens in nasal meatuses. PMID- 20027346 TI - Interferon status in children during acute respiratory infections. Therapy with interferon. AB - We studied interferon status in children during acute respiratory infections and found that it depends on etiology, course of the disease, and individual features of the organism. The efficiency of IFN inductor anaferon (pediatric formulation) and the possibility of its application in the therapy of children with acute respiratory infections were demonstrated. PMID- 20027347 TI - Anaferon (pediatric formulation) in prophylactics of acute respiratory viral infection in children. AB - Prophylactic treatment with anaferon (pediatric formulation) in children groups reduced total morbidity and incidence of acute respiratory viral infections and shortened the duration of fever, intoxication, and catarrhal syndromes. No allergic and other reactions caused by administration of the preparation were noted. PMID- 20027348 TI - Acute viral infections with combined involvement of the respiratory and gastrointestinal tracts in children. Therapy with interferon. AB - We evaluated the percent of acute respiratory viral infections with gastrointestinal syndrome in the structure of morbidity in babies aging 6 months and elder. Therapeutic efficiency and safety of anaferon (pediatric formuation) as a component of complex therapy of acute respiratory viral infections with involvement of the gastrointestinal tract were proven; more rapid disappearance of all symptoms and improvement of the immune status parameters were demonstrated. PMID- 20027349 TI - Clinical and immunological efficiency of anaferon (pediatric formulation) in calicivirus infection in children. AB - Therapy with anaferon (pediatric formulation) during the acute period of calicivirus infection shortened the duration of the main symptoms of the disease and period of virus release. Changes in the immunological status included increased production of IgA and IgM and activation of IFN-alpha synthesis. PMID- 20027350 TI - Evaluation of the efficiency and safety of anaferon (pediatric formulation) in the treatment of chickenpox in children. AB - Addition of anaferon (pediatric formulation) to the therapy of chickenpox patients led to more rapid disappearance of the main symptoms and alleviated the course of the disease. The safety of the preparation is confirmed by the absence of undesirable events and stability of laboratory indexes against the background of therapy. PMID- 20027351 TI - Application of ultralow doses of antibodies to interferon-gamma in complex therapy of bacterial infections and prophylaxis of bacterial complications. AB - Comparative placebo-controlled clinical trials on the efficiency and safety of ultralow doses of antibodies to human IFN-gamma (anaferon pediatric formulation and anaferon) and prophylaxis of bacterial complication showed that administration of these preparations in complex therapy of bacterial infection reduced the incidence of bacterial complications of viral infections and considerably decreased the duration of the main clinical symptoms of the disease. PMID- 20027352 TI - Efficiency of anaferon in complex therapy of genital herpes. AB - We studied clinical efficiency and IFN-inducing activity of anaferon in chronic recurrent genital herpes with high incidence of relapses. The use of anaferon in complex therapy reduced the duration of intoxication symptoms and local symptoms, shortened the duration of the relapse, activated expression of IFN-gamma mRNA, and improved IFN-gamma-producing capacity of blood leukocytes. PMID- 20027353 TI - Epidemiology and therapy of chronic herpes virus infection with genital herpes manifestations. AB - Anaferon-supplemented complex therapy of chronic recurrent genital herpes reduced clinical symptoms of the disease and normalized the content of immunocompetent cells. Changes in the level of immunocompetent cells depended on the severity of genital herpes. PMID- 20027354 TI - Clinical study of the efficiency and safety of afala in patients with benign prostatic hyperplasia. AB - The use of afala in patients with benign prostatic hyperplasia and moderate urination disturbances reduced the symptoms of the disease, improved urodynamic parameters, and increased quality of life. Clinical efficiency of afala was comparable with the efficiency of Serenoa repens extract (reference preparation). PMID- 20027355 TI - Experience of long-term afala treatment in benign prostatic hyperplasia. AB - The use of ultralow doses of antibodies to prostate-specific antigen (afala) for long-term treatment of benign prostatic hyperplasia in patients with moderate symptoms rapidly and effectively reduces irritative and obstructive symptoms, significantly decreases residual urine volume, and increases the rate of urination. Afala therapy is indicated for patients with stage I-II benign prostatic hyperplasia of moderately pronounced symptoms. PMID- 20027356 TI - Clinical use of afala in the therapy of chronic prostatitis. AB - The efficiency of afala in the therapy of chronic abacterial prostatitis was demonstrated. The preparation considerably improved urodynamics, reduced pain syndrome and inflammation, decreased prostate volume, improved patient's quality of life, and restored microcirculation in the prostate. No changes in blood, urine, and ECG parameters were noted. No side effects requiring afala withdrawal were recorded. PMID- 20027357 TI - Efficiency of afala in complex therapy of patients with chronic prostatitis. AB - The use of afala in complex treatment of patients with chronic prostatitis improves the efficiency of therapy and prolongs its positive results. The preparation is well tolerated, has no contraindication, and can be combined with other drugs. Afala is indicated for patients with stages II and IIIa chronic prostatitis from the first day of therapy. PMID- 20027358 TI - Prophylactics of erectile dysfunction in patients with metabolic syndrome. AB - For prophylactics of erectile dysfunction in patients with metabolic syndrome, all unfavorable life-style factors should be excluded and all metabolic disturbances should be corrected. This will reduce the severity and stabilize the course of metabolic syndrome. Administration of special drugs is required for prevention of erection disturbances. PMID- 20027359 TI - Efficiency of combined therapy with impaza and type 5 phosphodiesterase inhibitors in prophylactics of posttraumatic erectile dysfunction. AB - The use of combined pharmacotherapy with impaza and type 5 phosphodiesterase inhibitors helps to prevent and/or considerably reduce the risk of posttraumatic erectile dysfunction in men with traumas and strictures of the urethra. This therapy is indicated at early terms after trauma of the urethra for prophylactics of erectile dysfunction. PMID- 20027360 TI - Evaluation of the efficiency and safety of combined treatment with impaza and nitrates in CHD patients with erectile dysfunction. AB - The safety of combined administration of ultralow doses of antigens to endothelial NO synthase (impaza) and nitrates for the treatment of erectile dysfunction in CHD patients was evaluated in an open non-comparative clinical trial. The efficiency and safety of impaza and the possibility of its administration to patients receiving nitrates were demonstrated. PMID- 20027361 TI - Use of impaza in the treatment of erectile dysfunction in patients with essential hypertension and CHD. AB - Impaza treatment of erectile dysfunction in patients with essential hypertension and CHD receiving cardiotropic therapy improved erectile function and increased reserve circulation. Addition of impaza to the treatment protocol in cardiological patients considerably increased perfusion and reduced the content of desquamated epitheliocytes. No side effects of impaza were noted. PMID- 20027362 TI - The use of ultralow doses of antibodies to C-terminal fragment of angiotensin II AT1 receptor (kardos) in the therapy of arterial hypertension. AB - Kardos monotherapy allows attaining the target levels of systolic and diastolic blood pressure in patients with high-risk and very-high-risk hypertension. We demonstrated excellent tolerability of the preparation in combination with reliable blood pressure decrease over 24 h, during day and night hours. PMID- 20027363 TI - Pharmacodynamics of kardos administered as monotherapy and in combination with hypothiazide and enalapril in grade I-II arterial hypertension. AB - Therapy with kardos produced an antiihypertensve effect in patients with grade I II arterial hypertension. This antiihypertensve effect was considerably potentiated, when kardos was administered in combination with enalapril. PMID- 20027364 TI - The use of tenoten (pediatric formulation) in the therapy of chronic tension-type headaches in children. AB - Tension-type headaches are the most common type of headaches in children. No effective treatment for this disease is developed yet. After treatment with tenoten, the number of days with headache in 9-17-years-old children decreased by 40.9%, the mean headache intensity score decreased to 1. PMID- 20027365 TI - The use of tenoten and tenoten (pediatric formulation) as a drug for premedication in adults and children during outpatients dentist visit. AB - The use of tenoten and tenoten (pediatric formulation) for premedication in adults and children before dental treatment reduces patient anxiety; the effect is comparable with that of classical anxiolytics and is not accompanied by side effects. These preparations can be recommended for wide use in dentistry. PMID- 20027366 TI - Tenoten in the therapy of anxious disturbances in patients with essential hypertension and coronary heart disease. AB - The study demonstrated high anxiolytic activity of tenoten, which was not inferior to the anxiolytic effect of grandaxin. The positive changes persisted after termination of treatment in the tenoten group (but not in grandaxin group). Tenoten can be recommended for the treatment of patients with cardiovascular diseases associated with neurotic disturbances. PMID- 20027367 TI - Clinical effects of anxiolytic preparation tenoten in complex therapy of essential hypertension. AB - Addition of modern daily anxiolytic tenoten to complex therapy of patients with arterial hypertension improves the efficiency of treatment and reduces anxiety, which accelerates the development of hypotensive effect. Tenoten can be recommended for the treatment of anxiety symptoms in patients with arterial hypertension. PMID- 20027368 TI - The use of tenoten preparation in complex therapy of hypomotoric biliary dyskinesia. AB - A positive effect of tenoten on the course of biliary dyskinesia in patients with anxiety and depressive disorders was demonstrated. Tenoten can be recommended for the treatment of functional cholangio-pancreatoduodenal motility disturbances. PMID- 20027369 TI - Efficiency of antianxiety preparation tenoten used in complex therapy of patients with Helycobacter pylory-associated ulcer disease of the duodenum. AB - Tenoten reduces anxiety in patients with ulcer disease of the duodenum, promotes pain control, and induces no side effects. Tenoten can be recommended for active use in complex therapy of patients with ulcer disease. PMID- 20027370 TI - Tenoten in the therapy of patients with moderate cognitive impairment. AB - Tenoten is a new anxiolytic and antidepressant based on antibodies to brain specific protein S-100B. Experimental studies demonstrated the effect of tenoten on mechanisms of neuronal plasticity and manifestations of higher nervous activity. Tenoten is clinically comparable with amitryptiline, sertraline, and phenazepam, but does not produce potent sedative relaxation effect typical of these drugs. The study demonstrated considerable improvement of the control over brain frontal compartment effector functions. Tenoten is recommended not only at the stage of moderate cognitive impairment, but also in manifest cerebrovascular pathologies characterized by pronounced impairment of the regulatory functions of the frontal compartments of the brain. PMID- 20027371 TI - Integration column: microwell arrays for mammalian cell culture. AB - Microwell arrays have emerged as robust and versatile alternatives to conventional mammalian cell culture substrates. Using standard microfabrication processes, biomaterials surfaces can be topographically patterned to comprise high-density arrays of micron-sized cavities with desirable geometry. Hundreds to thousands of individual cells or cell colonies with controlled size and shape can be trapped in these cavities by simple gravitational sedimentation. Efficient long-term cell confinement allows for parallel analyses and manipulation of cell fate during in vitro culture. These live-cell arrays have already found applications in cell biology, for example to probe the effect of cell colony size on embryonic stem cell differentiation, to dissect the heterogeneity in single cell proliferation kinetics of neural or hematopoietic stem/progenitor cell populations, or to elucidate the role of cell shape on cell function. Here, we highlight the key applications of these platforms, hopefully inspiring biologists to apply these systems for their own studies. PMID- 20027373 TI - Microfluidic hydrodynamic cellular patterning for systematic formation of co culture spheroids. AB - This paper describes a microfluidic method to form co-culture spheroids of various geometries and compositions in order to manipulate cell-cell interaction dynamics. The cellular patterning is performed in a two-layered microfluidic device that sandwiches a semi-porous membrane so that flow occurs from the top channel through the membrane to the bottom channel. Arbitrary cellular arrangements are enabled by regulating the geometric features of the bottom channel so that as culture media drains, the flow hydrodynamically focuses (aggregates) cells onto the membrane only over the regions of the bottom channel. Furthermore, when the top channel has multiple inlets, cells can be seeded in adjacent laminar streams, allowing different cell types to be patterned simultaneously in well defined spatial arrangements. Interestingly, the initial cell positioning of certain cell types can result in two juxtaposed non concentric "Janus" spheroids, rather than homogeneous mixtures or layered shell structures. Therefore, the initial position of cells prior to aggregation can influence the final configuration within a co-culture spheroid. When Janus spheroids were constructed from mouse embryonic stem (mES) cells and hepatocytes, the mES cells differentiated in a spatially distinct pattern dictated by the position of the hepatocytes. This contrasts with uniform mES differentiation observed when co-culture spheroids are formed by the conventional method of randomly mixing the two cell types. This cellular patterning method opens new possibilities for understanding and manipulating interactions between different cell types in 3D. PMID- 20027374 TI - The dual nature of extracellular ATP as a concentration-dependent platelet P2X1 agonist and antagonist. AB - Patient groups subject to higher occurrence of stroke (e.g., people with diabetes, cystic fibrosis, pulmonary hypertension) have reduced release of ATP from their erythrocytes (ERYs) when subjected to flow-induced deformation or pharmacological stimuli. These same groups also have platelets that are more adhesive in comparison to controls. Here we show platelet aggregation, and inhibition of that aggregation, is affected by free Ca(2+) entering the platelet through the ATP-gated P2X1 receptor. The addition of ATP (10 microM) increased the platelet NO by 26.7 +/- 7.7%. This value was decreased significantly to below basal levels in the presence of NF 449 (p < 0.001), an inhibitor of the P2X1 receptor on the platelet. Aggregation profiles measured in the presence of ATP revealed that when the P2X1 receptor was blocked, or when the measurements were performed in Ca(2+) free buffer, platelet aggregation was nearly eliminated. Our findings employing standard aggregation measurements suggest that ATP behaves as a platelet inhibitor below 1.6 x 10(-19) moles ATP per platelet; however, above this value, ATP behaves as a platelet activator. These findings suggesting a dual nature of ATP with regard to platelet behavior were confirmed by passing platelets over endothelial cells that were coated in the channels of a microfluidic device. Importantly, it was determined that ERY-derived ATP release was a major determinant of platelet adhesion to the endothelium. These findings may have implications in anti-platelet drug design as most current therapies focus on the inhibition of P2Y-type receptors. Moreover, through the use of microfluidic technologies, we have provided in vitro evidence for a possible relationship between ERY properties and platelet behavior in vivo. PMID- 20027372 TI - Crosslinking of cell-derived 3D scaffolds up-regulates the stretching and unfolding of new extracellular matrix assembled by reseeded cells. AB - Elevated levels of tissue crosslinking are associated with numerous diseases (cancer stroma, organ fibrosis), and also eliminate the otherwise remarkable clinical successes of tissue-derived scaffolds, instead eliciting a foreign body reaction. Nevertheless, it is not well understood how the initial physical and biochemical properties of cellular microenvironments, stem cell niches, or of 3D tissue scaffolds guide the assembly and remodeling of new extracellular matrix (ECM) that is ultimately sensed by cells. Here, we incorporated FRET-based mechanical strain sensors, either into cell-derived ECM scaffolds or into the fibronectin (Fn) matrix assembled by reseeded fibroblasts, and demonstrated the following. Cell-generated tensile forces change the conformation of Fn in both 3D scaffolds and new matrix over time. The time course by which new matrix fibers are stretched by reseeded cells is accelerated by scaffold crosslinking. Importantly, stretching Fn fibers increases their elastic modulus (rigidity) and alters their biochemical display. Regulated by Fn fiber unfolding, more soluble Fn binds to the native than to the crosslinked scaffolds. Additionally, matrix assembly of fibroblasts is decreased by scaffold crosslinking. Taken together, scaffold crosslinking has a multifactorial impact on the microenvironment that reseeded cells assemble and respond to, with far-reaching implications for tissue engineering and disease physiology. PMID- 20027375 TI - Dynamics of virus spread in the presence of fluid flow. AB - The dynamics of viral infection spread, whether in laboratory cultures or in naturally infected hosts, reflects a coupling of biological and physical processes that remain to be fully elucidated. Biological processes include the kinetics of virus growth in infected cells while physical processes include transport of virus progeny from infected cells, where they are produced, to susceptible cells, where they initiate new infections. Mechanistic models of infection spread have been widely developed for systems where virus growth is coupled with transport of virus particles by diffusion, but they have yet to be developed for systems where viruses move under the influence of fluid flows. Recent experimental observations of flow-enhanced infection spread in laboratory cultures motivate here the development of initial continuum and discrete virus particle models of infection spread. The magnitude of a dimensionless group, the Damkohler number, shows how parameters that characterize particle adsorption to cells, strain rates that reflect flow profiles, and diffusivities of virus particles combine to influence the spatial pattern of infection spread. PMID- 20027380 TI - Design issues for socially intelligent user interfaces. A discourse analysis of a data-to-text system for summarizing clinical data. AB - OBJECTIVES: This study aims to demonstrate the usability of discourse analyses as a means of evaluating medical informatics systems by examining one particular computer-based data-to-text system for delivering neonatal health care information. METHODS: Six textual summaries of clinical information, three produced by human clinicians and three by the data-to-text system, were subjected to fine-grain discourse analysis. Analysis was performed 'blind' on all six textual summaries. Analysis focused on the identification of lexical items and on the potential effects of these items on users of these clinical information summaries. RESULTS: Results showed that there were clear differences between human- and system-generated clinical summaries, with human clinicians providing better narrative flow and textual detail. The data-to-text system successfully produced textual summaries although it fell short of human abilities. CONCLUSIONS: These results indicate potential future improvements to the system. Discourse analysis as used here may offer significant advantages in evaluating and developing similar medical informatics systems. PMID- 20027381 TI - A new prior for bayesian anomaly detection: application to biosurveillance. AB - OBJECTIVES: Bayesian anomaly detection computes posterior probabilities of anomalous events by combining prior beliefs and evidence from data. However, the specification of prior probabilities can be challenging. This paper describes a Bayesian prior in the context of disease outbreak detection. The goal is to provide a meaningful, easy-to-use prior that yields a posterior probability of an outbreak that performs at least as well as a standard frequentist approach. If this goal is achieved, the resulting posterior could be usefully incorporated into a decision analysis about how to act in light of a possible disease outbreak. METHODS: This paper describes a Bayesian method for anomaly detection that combines learning from data with a semi-informative prior probability over patterns of anomalous events. A univariate version of the algorithm is presented here for ease of illustration of the essential ideas. The paper describes the algorithm in the context of disease-outbreak detection, but it is general and can be used in other anomaly detection applications. For this application, the semi informative prior specifies that an increased count over baseline is expected for the variable being monitored, such as the number of respiratory chief complaints per day at a given emergency department. The semi-informative prior is derived based on the baseline prior, which is estimated from using historical data. RESULTS: The evaluation reported here used semi-synthetic data to evaluate the detection performance of the proposed Bayesian method and a control chart method, which is a standard frequentist algorithm that is closest to the Bayesian method in terms of the type of data it uses. The disease-outbreak detection performance of the Bayesian method was statistically significantly better than that of the control chart method when proper baseline periods were used to estimate the baseline behavior to avoid seasonal effects. When using longer baseline periods, the Bayesian method performed as well as the control chart method. The time complexity of the Bayesian algorithm is linear in the number of the observed events being monitored, due to a novel, closed-form derivation that is introduced in the paper. CONCLUSIONS: This paper introduces a novel prior probability for Bayesian outbreak detection that is expressive, easy-to-apply, computationally efficient, and performs as well or better than a standard frequentist method. PMID- 20027382 TI - Towards a traceable clinical guidelines application. A model-driven approach. AB - OBJECTIVES: The goal of this research is to provide an overall framework to enable model-based development of clinical guideline-based decision support systems (GBDSSs). The automatically generated GBDSSs are aimed at providing guided support to the physician during the application of guidelines and automatically storing guideline application data for traceability purposes. METHODS: The development process of a GBDSS for a guideline is based on model driven development (MDD) techniques which allow us to carry out such a process automatically, making development more agile and saving on human resource costs. We use UML Statecharts to represent the dynamics of guidelines and, based on this model, we use a MDD-based tool chain to generate the guideline-dependent components of each GBDSS in an automatic way. In particular, as for the traceability capabilities of each GBDSS, MDD techniques are combined with database schema mappings for metadata management in order to automatically generate the GBDSS-persistent component as one of the main contributions of this paper. RESULTS: The complete framework has been implemented as an Eclipse plug-in named GBDSSGenerator which, starting from the statechart representing a guideline, allows the development process to be carried out automatically by only selecting different menu options the plug-in provides. We have successfully validated our overall approach by generating the GBDSS for different types of clinical guidelines, even for laboratory guidelines. CONCLUSIONS: The proposed framework allows the development of clinical guideline-based decision support systems in an automatic way making this process more agile and saving on human resource costs. PMID- 20027383 TI - Reviews in epidemiology: diversity in the research agenda. PMID- 20027384 TI - Non classical risk factors for gestational diabetes mellitus: a systematic review of the literature. AB - Age, obesity and family history of diabetes are well known risk factors for gestational diabetes mellitus. Others are more controversial. The objective of this review is to find evidence in the literature that justifies the inclusion of these other conditions among risk factors. The MEDLINE, Cochrane, LILACS and Pan American Health Organization databases were searched, covering articles dating from between 1992 and 2006. Keywords were used in combination (AND) with gestational diabetes mellitus separately and with each one of the risk factors studied. The methodological quality of the studies included was assessed, resulting in the selection of 41 papers. Most studies investigating maternal history of low birth weight, low stature, and low level of physical activity have found positive associations with gestational diabetes mellitus. Low socioeconomic levels, smoking during pregnancy, high parity, belonging to minority groups, and excessive weight gain during pregnancy presented conflicting results. Publication bias cannot be ruled out. Standardization of techniques, cutoff points for screening and diagnosis, as well as studies involving larger sample sizes would allow future meta-analyses. PMID- 20027385 TI - Adverse drug events in hospitals: a systematic review. AB - The objective of this study was to evaluate studies on the occurrence of adverse drug events (ADEs) in hospitals in order to learn about their frequency and characteristics, comparing the methods for identifying them and the various definitions. A search was conducted on MEDLINE and identified studies published from 2000 to 2009. Inclusion criteria were: studies in populations not selected for specific diseases or drugs and ADEs that occurred during hospitalization. Twenty-nine studies were selected, displaying multiple sources of heterogeneity, including differences in the study populations, surveillance techniques, definitions of ADEs, and indicators. The proportion of patients with ADEs ranged from 1.6% to 41.4% of inpatients and the rates ranged from 1.7 to 51.8 events/100 admissions. A considerable share of these events could have been avoided. The findings show that ADEs in inpatients are a public health problem. However, further studies are needed to monitor these adverse events in order to effectively promote safe drug use. PMID- 20027386 TI - Risk factors for healthcare-associated infection in pediatric intensive care units: a systematic review. AB - A systematic review of observational studies on risk factors for healthcare associated infection in pediatric Intensive Care Units (ICU) was carried out. Studies indexed in MEDLINE, LILACS, Cochrane, BDENF, CAPES databases published in English, French, Spanish or Portuguese between 1987 and 2006 were included and cross references added. Key words for search were 'cross infection' and 'Pediatric Intensive Care Units' with others sub-terms included. 11 studies were selected from 419 originally found: four studies had healthcare-associated infection as the main outcome without a specific site; three articles identified factors associated with lower respiratory tract infection (pneumonia or tracheitis); three articles were concerned with laboratory-confirmed bloodstream infection; and a single retrospective study analyzed urinary tract infection. The production of evidence on risk factors Paediatric ICU has not kept up the same pace of that on adult - there are few studies with adequate design and statistical analysis. The methodological diversity of the studies did not allow for a summarized measurement of risk factors. PMID- 20027387 TI - Assessing patient adherence to chronic diseases treatment: differentiating between epidemiological and clinical approaches. AB - This review discusses the concepts and methods for assessing patient adherence to treatment, as applied to both epidemiological and clinical approaches within real health care practices. For the epidemiological approach, the assessment must be as accurate as possible. Self-reported questionnaires are the most feasible option in most circumstances, but most demonstrate low sensitivity combined with high specificity. We suggest that self-reported outcomes, where feasible, can increase the sensitivity for non-adherence of these questionnaires. In the clinical approach an accurate distinction between adherents and non-adherents is less useful. For the health provider, it is more important to be aware of the particular situation that each patient is currently experiencing with his/her treatment. Self-reported questionnaires applied in clinical settings can help the health provider to form an objective opinion. In any event, the patient-provider dialogue is still the best approach to assess patient adherence as well as to deliver good care. PMID- 20027388 TI - Impact of methodology on the results of economic evaluations of varicella vaccination programs: is it important for decision-making? AB - This study aims to review the literature on economic evaluation of childhood varicella vaccination programs and to discuss how heterogeneity in methodological aspects and estimation of parameters can affect the studies' results. After applying the inclusion criteria, 27 studies published from 1980 to 2008 were analyzed in relation to methodological differences. There was great heterogeneity in the perspective adopted, evaluation of indirect costs, type of model used, modeling of the effect on herpes zoster, and estimation of vaccine price and efficacy parameters. The factor with the greatest impact on results was the inclusion of indirect costs, followed by the perspective adopted and vaccine price. The choice of a particular methodological aspect or parameter affected the studies' results and conclusions. It is essential that authors present these choices transparently so that users of economic evaluations understand the implications of such choices and the direction in which the results of the analysis were conducted. PMID- 20027389 TI - Physical activity in Brazil: a systematic review. AB - The purpose of this study, based on a systematic literature review, was to describe the prevalence of physical activity (or inactivity) in the Brazilian population. The databases consulted were: LILACS, SciELO, MEDLINE, Web of Science, and the Google Scholar portal. The terms 'physical activity', 'physical exercise', 'physical inactivity', 'sedentary' 'Brazil', and 'Brazilian' were used in the search. Overall, 47 studies (all cross-sectional) with random samples were found, and in 26 studies physical activity was the main variable. Only two studies were published before the year 2000, as compared to 12 in 2008 alone. The studies were heavily concentrated in the South and Southeast of Brazil, and there were few studies on physical activity in children and adolescents. In all the studies, physical activity was measured subjectively, mainly with questionnaires, and the most widely studied domain was leisure time. The criteria for defining physical activity varied widely, as did prevalence. The study highlighted the need for standardization of instruments, criteria, and nomenclature in epidemiological studies on physical activity. PMID- 20027390 TI - Childhood poverty and abdominal obesity in adulthood: a systematic review. AB - Adverse socioeconomic conditions in childhood can have lasting effects on health, but evidence is lacking from prospective studies concerning the effects of early poverty on abdominal obesity in adulthood. Cross-sectional studies in adults from middle and high-income countries show that current socioeconomic status is inversely related to obesity in women, but the pattern in men is not consistent. A systematic review was undertaken to assess the influence of early socioeconomic status on waist circumference, hip circumference, and waist-hip ratio in adulthood. Thirteen relevant articles were located (five cross-sectional and eight cohort), including only one from a middle-income country and the remainder from high-income settings. In all the studies, childhood poverty was associated with higher levels of abdominal obesity in women. In men, the associations were weaker, and no clear pattern emerged. PMID- 20027391 TI - Magnetic fields and acute lymphoblastic leukemia in children: a systematic review of case-control studies. AB - Leukemia incidence in children has increased worldwide in recent decades, particularly due to the rise in acute lymphoblastic leukemia. Studies have associated exposure to non-ionizing radiation generated by low frequency magnetic fields with childhood leukemia. The current article reviews the case-control studies published on this subject. Of 152 articles tracked in different databases, ten studies from North America, Asia, and Europe met the defined selection criteria, with patients diagnosed from 1960 to 2004. Methodological limitations were observed in these articles, including difficulties with the procedures for assessing exposure. An association may exist between exposure to low frequency magnetic fields and acute lymphoblastic leukemia in children, but this association is weak, preventing the observation of consistency in the findings. Future studies from a wider range of geographic regions should focus on the analysis of acute lymphoblastic leukemia, which is the subtype with the greatest impact on the increasing overall incidence of childhood leukemia. PMID- 20027392 TI - Association between stress and breast cancer in women: a meta-analysis. AB - The objective of the current meta-analysis was to verify the association between stressful life events and primary breast cancer incidence in women. A total of 618 studies from 1982-2007 were found in the PubMed, LILACS, and Cochrane Library databases. Methodological quality was evaluated according to the Downs & Black criteria. Eight studies were selected (six case-controls and two cohorts). The studies were grouped in three analyses, two of which based on the categories widowhood and divorce and the other based on self-rated intensity and frequency of stressful events. Relative risks were: widowhood 1.04 (95%CI: 0.75-1.44; p = 0.800); divorce 1.03 (95%: 0.72-1.48; p = 0.850); and intensity/frequency of stress 1.73 (95%CI: 0.98-3.05; p = 0.059). We conclude that stressful life events as a whole are not associated with risk of breast cancer in women. However, it is not possible to rule out high-intensity stress as a risk factor for breast cancer. PMID- 20027393 TI - Gender and incidence of functional disability in the elderly: a systematic review. AB - This systematic review aimed to assess the effect of gender on the incidence of functional disability in the elderly. The search for publications in the MEDLINE, EMBASE, and ProQuest electronic databases from January 1990 to March 2008 identified 879 publications, from which 21 cohort studies were selected that evaluated gender as an independent variable and incidence of functional disability in elderly as the outcome. Review of the studies found that after adjusting for socioeconomic, health, and social relations indicators, incidence of functional disability was similar between genders. The main risk factors for functional disability in the elderly, regardless of gender, were: lack of schooling, living in rental housing, chronic diseases, arthritis, diabetes, visual impairment, body mass index above 25, poor self-perceived health, cognitive impairment, depression, slow gait, sedentary lifestyle, tiredness while performing daily activities, and limited diversity in social relations. PMID- 20027395 TI - [Strabismus: from theory to practice, from concepts to its operational attainment]. AB - Some particular points concerning the study of strabismus are analyzed, specially, the difficulties for definitions and the different ways it can be conceived, besides the relativity of binocular positions interpretation, and the necessary referential elements for circumscribing such questions. Then, the concept of primary position of gaze is discussed as well as the conditions for its operational attainment, leading to the conclusion that it is materially impossible to achieve this 'zero' point, from which all other measurements of strabismus or ocular movements should be done. In sequence, the construction of multiple referential systems applying the quantification of strabismus are examined. The lack of consensual agreement about which should be used as the standard system causes elementary practical implications, as the lack of agreement about how to superimpose orthogonal prisms for the measurement of associated horizontal and vertical deviations. The functional asymmetry of ocular rotations and its exceptions are commented. The accuracy that the measurements of strabismus can be performed is analyzed, leading to the conclusion that fractional figures using the commonest angular unities are improper. At last, some technical problems related to such evaluations are also commented, as operational difficulties related to prism-diopter unity, concerning the use of prisms, and prismatic effects due to the use of conventional optical glasses. PMID- 20027396 TI - Study of ocular aberrations with age. AB - INTRODUCTION: Aging has various effects on visual system. Vision deteriorate, contrast sensitivity decreases and ocular aberrations apparently make the optical quality worse across the years. PURPOSE: To prospective evaluate ocular aberrations along the ages. METHODS: Three hundred and fifteen patients were examined, 155 were male (39.36%) and 160 were female (60.63%). Ages ranged from 5 to 64 year-old, the study was performed from February to November, 2004. Patients were divided into 4 age-groups according to IBGE (Instituto Brasileiro de Geografia e Estatistica) classification: 68 patients from 5 to 14 year-old, 55 patients from 15 to 24 year-old, 116 from 25 to 44 year-old and 76 from 45 to 67 year-old. All patients had the following characteristics: best corrected visual acuity > 20/25, emmetropia or spherical equivalent < 3.50 SD, refractive astigmatism < 1.75 CD on cycloplegic refraction, normal ophthalmologic exam and no previous ocular surgeries. This protocol was approved by Federal University of Sao Paulo Institutional Review Board. Total optical aberrations were measured by H-S sensor LadarWave Custom Cornea Wavefront System (Alcon Laboratories Inc, Orlando, FLA, USA) and were statistically analysed. Corneal aberrations were calculated using CT-View software Version 6.89 (Sarver and Associates, Celebration, FL, USA). Lens aberrations were calculated by subtraction. RESULTS: High-order (0.32 e 0.48 microm) and ocular spherical aberrations (0.02 e 0.26 microm) increased respectively in child and middle age groups. High order (0.27 microm) and corneal spherical aberrations (0.05 microm) did not show changes with age. Lens showed a statistically significant spherical aberration increase (from 0.02 to 0.22 microm). Vertical (from 0.10 to -0.07 microm) and horizontal coma (from 0.01 to -0.12 microm) presented progressively negative values with aging. CONCLUSION: High-order and spherical aberrations increased with age due to lens contribution. The cornea did not affect significantly changes observed on ocular aberrations. PMID- 20027397 TI - Comparison between retinal nerve fiber layer and macular thickness measured with OCT detecting progressive axonal loss following traumatic optic neuropathy. AB - PURPOSE: To compare the optical coherence tomography retinal nerve fiber layer and macular thickness measurements for detection of progressive axonal loss following acute traumatic optic neuropathy in a longitudinal study. METHODS: Three patients with unilateral traumatic optic neuropathy were evaluated sequentially after trauma. Macular and retinal nerve fiber layer thickness measurements were obtained using optical coherence tomography weekly for five weeks and around the twelfth week after trauma. RESULTS: All patients showed progressive macular and retinal nerve fiber layer thickness reduction. The mean retinal nerve fiber layer thickness on the first week was 114 microm and reduced sequentially over the first five weeks and was 46 microm on the twelfth week. For macular parameters, the mean average thickness on the first week was 248 microm and also reduced over the first five weeks and was 218 microm on the twelfth week. When compared to the initial measurement, macular thickness average reduction rate at the 12th week was 14% while peripapillary retinal nerve fiber layer thickness average reduction rate was 59%. CONCLUSIONS: Although both measurements reduce significantly after trauma, retinal nerve fiber layer thickness measurements show greater and faster retinal neural reduction if compared to macular thickness measurements in traumatic optic neuropathy. PMID- 20027398 TI - Antimicrobial efficacy assessment of multi-use solution to disinfect hydrophilic contact lens, in vitro. AB - PURPOSE: To evaluate the efficacy of disinfecting solutions in hydrophilic contact lenses (CL). METHODS: Two multi-use solutions denominated solution A (0.001% polyquaternium-1 and 0.0005% myristamidopropyl dimethylamine) and solution B (0.0001% polyaminopropyl biguanide) were used. The solutions were tested in hydrophilic contact lenses infected with Pseudomonas aeruginosa (ATCC27583), Staphylococcus epidermidis (ATCC1226), Klebsiella pneumoniae (ATCC13883), Staphylococcus aureus (ATCC25923) and Candida albicans (ATCC 10231) and the decrease in microorganisms growth after the hydrophilic contact lenses were cleaned with the respective solutions was verified. The manufacture's instructions were followed. RESULTS: A decrease of 90% of Pseudomonas aeruginosa, Staphylococcus epidermidis, Staphylococcus aureus, Candida albicans and a decrease 100% of Klebsiella pneumoniae was observed. CONCLUSION: The solutions decreased the amount of microorganisms tested. PMID- 20027399 TI - [Evaluation of the quality of donor corneas in relation to the age of donor and cause of death]. AB - PURPOSE: To analyze the quality of the corneas evaluated by slit lamp examination in the Eye Bank of the Hospital de Clinicas UFPR and its relation to donor age and cause of death. METHODS: Analysis of 492 corneas, evaluated in BTO HC-UFPR between August 2006 to August 2008. Each cornea was classified regarding the quality as: very good, good, regular and bad; and according to clarity, epithelial defect, stromal edema, Descemet's folds, stromal opacity, corneal scarring, endothelial density and guttata. RESULTS: The mean donor age was 42.74 years (sd=17.77 years). Among the causes of death, trauma was the most common with 46.18%, followed by cardiovascular causes with 41.86%. In relation to the cornea, 57.11% were classified as 'good', followed by 20.73% as 'regular', 16.87% as 'very good' and 5.28% as 'bad'. With the application of statistical tests based on ordinal regression model, trauma deaths corneas tend to be better, as well as those of younger patients. CONCLUSIONS: Statistically, the corneas from younger donors tend to have better graduations in the assessment, as well as the corneas from donors dead by trauma that tend to have better quality when compared to corneas from donors dead by cardiovascular and other systemic diseases. PMID- 20027400 TI - Visual performance after toric IOL implantation in patients with corneal astigmatism. AB - PURPOSE: To analyze visual acuity without correction and rotational stability outcomes following toric IOL implantation. METHODS: Prospective study of 20 eyes of 13 patients that underwent phacoemulsification surgery indicated for cataract associated with regular keratomeric astigmatism, symmetrical, ranging from 1 to 4 diopters. Best corrected visual acuity, refraction, keratometry and computed topography were performed preoperatively. The calculation of cylindrical lens power and its placement were determined by the manufacturer. All lenses were implanted in the capsular bag by the same surgeon. The patients were examined by a second independent observer, at 1st, 10th, 20th, 30th, and 60th postoperative day. RESULTS: Visual acuity without correction ranged between 20/15 and 20/40. One eye achieved 20/15 (5%), 4 eyes 20/20 (20%), 6 eyes 20/25 (30%), 7 eyes 20/30 (35%) and 2 eyes 20/40 (10%). Best corrected visual acuity ranged between 20/15 and 20/40; two eyes with 20/15 (10%), 9 eyes 20/20 (45%), 7 eyes 20/25 (35%), 1 eye 20/30 (5%) and 1 eye 20/40 (5%). It is important to remember that the average spherical refraction was -0.05 SD (ranging from -0.50 to +0.75 SD). The mean cylindrical refraction was -0.63 CD ranging from -0.50 to -1.25 CD. The IOL rotation in this study had an average of 3.2 masculine to 30 masculine, ranging from 0 masculine of rotation to a maximum of 13 masculine; 7 lenses (35%) suffered no rotation, 9 lenses (45%) suffered rotation between 1 masculine to 5 masculine, 3 lenses (15%) had rotation between 6 masculine to 10 masculine, and ultimately 1 lens (5%) had rotation between 11 masculine to 15 masculine. There was no significant rotation after the 30th postoperative day. DISCUSSION: The average of rotation of the IOL was 3.2 masculine, where 95% of IOLs presented rotation less than or equal to 10 masculine what means a very good rotational stability. In daily practice, a good visual acuity is directly related to IOL rotational stability and refractive predictability. PMID- 20027401 TI - Alternative technique for reducing compound waste during intravitreal injections. AB - PURPOSE: To describe an intravitreal injection technique using a commercially available 29-gauge insulin needle syringe (29GN syringe) and a 21-gauge (G) needle, comparing compound waste associated with this technique application and the one described in ranibizumab (Lucentis) kit instructions. METHODS: Ten 0.3 ml doses of distilled water were aspirated using the 29GN syringe and 21G needle (PT technique), and another ten equal doses were aspirated employing the sterilized Lucentis kit (LK technique), which contains a 1ml tuberculin syringe, a 18G needle for compound aspiration and a 30G needle for intravitreal injection. For aspiration using the PT technique, a 21G needle is attached over a 29GN syringe. After compound aspiration, the 21G needle is removed and intravitreal injection is performed using the 29G needle. Using a precision balance, the aspiration needles (21G for PT; 18G for LK) were weighed before and after water aspiration and the syringe-needle complexes for injection (29GN for PT; 30G for LK) were weighed before aspiration and after emptying them. The volumes left in the aspiration needles and needle-syringe complexes were estimated by the difference in weight in grams, which were converted to millilitres. RESULTS: The mean (+/ SD) residual volume (ml) of aspiration needles (21G for PT; 18G for LK) was significantly lower with PT technique (0.0034 +/- 0.0016) when compared to LK tech nique (0.0579 +/- 0.0011) (p<0.01). The mean (+/-SD) residual volume (ml) of syringe-needle complexes was significantly lower with PT technique (0.0056 +/- 0.0011) than with LK (0.0906 +/- 0.003 ml) (p<0.01). CONCLUSION: The proposed technique is a reasonable alternative for minimizing medication loss during intravitreal injection procedures. PMID- 20027402 TI - Amblyopia after unilateral infantile cataract extraction after six weeks of age. AB - PURPOSE: To determine interocular grating acuity difference in children treated for unilateral infantile cataract. METHODS: A group of 27 children previously treated for unilateral infantile cataract, had their monocular visual acuity measured by sweep visual evoked potentials. Interocular grating acuity difference was calculated as the absolute subtraction of monocular acuity scores. Lens status, opacity severity and eye alignment were considered for analysis. RESULTS: Mean interocular grating acuity difference obtained from unilateral cataract patients was 0.58 +/- 0.20 logMAR. This result was significantly larger than 0.10 logMAR used as normative data. Children with severe opacities had a more pronounced amblyopia than the moderate ones. No significant correlation between amblyopia and strabismus or aphakia was found. CONCLUSIONS: Interocular acuity difference in this group of unilateral congenital cataract was more pronounced than previous reports, mainly because of delay in diagnosis, surgery and optical correction. PMID- 20027403 TI - [Experimental study of vitreoretinal proliferation inhibition by the use of hypericin]. AB - PURPOSE: To produce proliferative vitreoretinopathy (PVR) in an animal ocular trauma model. To evaluate the inhibition of (PVR) emergence and progression by hypericin. METHODS: Experimental Study. Nineteen pigmented male adult rabbits weighing between 2,000 and 3,000 grams were used in this study. All of them were submitted to trauma model with dispase and retinal diathermy to induce PVR membranes formation. They were randomly assigned to receive hypericin (10 microM in 0.1 ml) or saline solution (0.1 ml) as placebo. They were evaluated clinically in the seventh, fourteenth, twenty-first and twenty-eighth postoperative days with indirect ophthalmoscopy and digital color retinography. The PVR degree was classified according to Hida (0 to 7). RESULTS: Membranes formation was present in 79% of the eyes; being 100% in the eyes of placebo group and 60% in the eyes of treatment group (hypericin). The comparison between PVR phases averages within the groups showed a statistically significant difference between the two groups, with a p value of 0.0321 for Wilcoxon test. CONCLUSIONS: The trauma model with dispase and retinal diathermy produces vitreoretinal membranes. Hypericin was considered effective in PVR emergence and progression decrease. PMID- 20027404 TI - Intravitreal bevacizumab in pigmented rabbit eyes: histological analysis 90 days after injection. AB - PURPOSE: To evaluate bevacizumab toxicity in neurosensorial retina and retinal pigment epithelium in pigmented rabbit eyes by means of histological studies. METHODS: Thirty eyes of fifteen rabbits were distributed into three groups: sham group (S), that received a 0.1 ml balanced saline solution (BSS) intravitreal injection (10 eyes); group 1, that received a 1.25 mg (0.1 ml) bevacizumab intravitreal injection (10 eyes); and group 2, that received a 2.5 mg (0.1 ml) bevacizumab intravitreal injection (10 eyes). Rabbits were sacrificed 90 days after the procedure and both eyes of each rabbit were enucleated. A histological examination of neurosensorial retina and retinal pigmented epithelium (RPE) was performed. Its morphological features and layer thickness were also analyzed. RESULTS: No histological differences in neurosensorial retina or in retinal pigmented epithelium were found and layer thickness did not differ significantly between balanced saline solution-injected eyes and bevacizumab-injected eyes. CONCLUSION: After a 90-day follow-up period, a single 1.25 or 2.5 mg bevacizumab intravitreal injection did not lead to toxic damage in the neurosensorial retina and retinal pigment epithelium of pigmented rabbit eyes, and it appears to be a safe procedure for retinal neovascular diseases. PMID- 20027405 TI - [Comparison of the analgesic effect between 90 mg etoricoxib and dipyrone after exeresis of primary pterygium with conjunctival autograft]. AB - PURPOSE: To compare the analgesic effect between dipyrone, 90 mg etoricoxib, and placebo after excision of primary pterygium with conjunctival autograft. METHODS: Prospective, randomized, double-masked clinical trial. Three groups of 26 patients (one eye per patient) were submitted to surgery and received the study drugs for five days after surgery. A scale of pain was used, graduated from zero to ten, for patient evaluation in the first, third and fifth postoperative days. The pain was classified as absent (zero), mild (1 to 3), moderate (4 to 7) and severe (8 to 10). Statistical analysis was performed with the SPSS, version 11.5. RESULTS: A statistically significant difference was found between etoricoxib and dipyrone in the first and third postoperative days (p=0.001 and p=0.01; respectively). Etoricoxib was superior to placebo only in the first postoperative day (p=0.04). There was no significance in the comparison between dipyrone and placebo. CONCLUSIONS: Analgesia of etoricoxib was superior to placebo in the first postoperative day and to dipyrone in the third and fifth days after excision of primary pterygium with conjunctival autograft. There was no significant difference between dipyrone and placebo in all time points. PMID- 20027406 TI - [Oral sedation with midazolam in blepharoplasty]. AB - PURPOSE: To evaluate the safety and usefulness of the use of oral sedation with midazolam (15 mg) in patients submitted to blepharoplasty. METHODS: Randomized double-blind prospective study of 42 patients (surgical risk ASA I and II) divided into three groups of 14 patients each: Group M (midazolam 15 mg), group P (placebo) and group SM (no medication). All patients were evaluated according to the degree of sedation and pain during surgery and the variation of anxiety between the preoperative and intraoperative period, arterial pressure (systolic SAP and diastolic-DAP), respiratory frequency (RF) and pulsation. RESULTS: Unifatorial variance analysis with Tukey test demonstrated that the use of midazolam provoked a significant SAP and RF reduction during the intraoperative period. These effects were not pronounced and were accompanied by a reduction of pain perception and anxiety and mild sedation. CONCLUSIONS: Oral sedation with midazolam in patients that had undergone eyelid surgical procedures is safe and easy to perform with minimal systemic effects. PMID- 20027407 TI - [Balloon-catheter dilatation (dacrioplasty) for congenital nasolacrimal duct obstruction treatment]. AB - PURPOSE: To verify the results of balloon-catheter dilatation for the treatment of congenital lacrimal duct obstruction. To the best of author's knowledge this procedure has not been previously used in Brazil and deserves scientific studies. METHODS: Children between 2 and 5 years old, with congenital lacrimal duct obstruction, were treated with balloon-catheter dilatation (Lacricath). Catheter was introduced by the superior lacrimal punctum, under general anesthesia. RESULTS: Twenty-seven eyes were treated and 23 presented total improvement of epiphora signs and symptoms (85.2% success rate). During a one year follow-up, all the procedures had good outcomes, without complications and all the children had 1 year of follow-up. CONCLUSION: Balloon-catheter dilatation is an efficient and safe procedure and might be used for congenital lacrimal duct obstruction treatment in children older than 2 years. PMID- 20027408 TI - [Evaluation of corneal tissue processed by a reference Eye Bank]. AB - PURPOSE: To evaluate the quality of the corneas processed by Sorocaba Eye Bank (BOS) - SP and transplanted out of the Ophthalmology Hospital of Sorocaba (HOS), as well as the corneas that had re-entered in the BOS, being transplanted or not in the HOS, during the year of 2007. METHODS: Ophthalmologists that transplanted corneas processed by BOS outside of the Ophthalmology Hospital of Sorocaba were contacted, as well as those who had used re-entried corneas, to collect the following information: period from eye enucleation until transplant, transparency of the donated tissue, time of transplant and primary failure. RESULTS: During the year of 2007, Sorocaba Eye Bank distributed three hundred and ninety-two corneas for outside HOS. From these, six had returned to BOS and had been transplanted in the Ophthalmology Hospital of Sorocaba. From those, none was rejected; however, two presented some opacity in the visual axis. After attempt to congregate information regarding the 386 transplanted corneas outside HOS, data of only 48 keratoplasty were available. The average time from the enucleation until the keratoplasty was of 5.5 days (1-13 days), and of postoperative average follow-up, 9.8 months (4-15 months). Three corneas had developed primary failure; three presented opacity in the visual axis; one presented infectious keratitis, requiring therapeutic keratoplasty; a patient died and another one lost follow-up. The other 39 corneas did not present any problems until the time this study was ended. CONCLUSION: Because of the difficulties to congregate informations of the patients transplanted in other hospitals, it is difficult to define the quality of the distributed corneas by Sorocaba Eye Bank. Thus, other analysis are necessary in order to define changes and new directions for future studies on selection and preservation of donated corneas. PMID- 20027409 TI - Intravitreal ranibizumab and bevacizumab for the treatment of nonsubfoveal choroidal neovascularization in age-related macular degeneration. AB - PURPOSE: To investigate the efficacy of vascular endothelial growth factor specific (VEGF) monoclonal antibodies in the treatment of choroidal neovascularization secondary to age-related macular degeneration (AMD) that does not extend beneath the foveal center (nonsubfoveal CNV). METHODS: The study design was a retrospective chart review of consecutive patients over a two-month period under active treatment with bevacizumab and/or ranibizumab for neovascular AMD. Patients with neovascularization within the macula that did not extend beneath the center of the foveal avascular zone, along with at least one large drusen (>125 micro) or many intermediate size (63-124 micro) drusen were included. Best corrected Snellen visual acuity and optical coherence tomography (OCT) analysis of the central macular thickness was recorded for each visit. Serial injections of bevacizumab and/or ranibizumab were administered until there was resolution of subretinal fluid clinically or by OCT. Data over the entire follow-up period were analyzed for overall visual acuity and OCT changes. All patients had follow-up since diagnosis of at least 6 months (mean=9.6 months). RESULTS: Of the thirteen included patients, eleven had reduction of retinal thickening in the area involved by the CNV. The remaining two patients did not have OCT data available but had no fluid or activity on clinical examination at last follow-up. One patient (8%) lost one line of vision; one (8%) remained stable, and eleven (84%) gained one or more lines of visual acuity. Three patients (23%) gained three or more lines. The average treatment outcome for all patients was a gain of 1.7 +/- 1.3 lines of Snellen acuity. Both therapeutic agents were effective, with an average gain of 1.6 +/- 0.6 lines for patients treated with bevacizumab, 1.5 +/- 1.9 lines gained for patients treated with ranibizumab and 2.5 +/- 0.7 lines gained in the two patients who received both agents over the course of their treatment. CONCLUSIONS: The use of intravitreal anti-VEGF agents for nonsubfoveal CNV in AMD is effective. Our results are comparable to published results from large-scale trials of anti-VEGF therapy for subfoveal CNV. Our data support the idea that bevacizumab or ranibizumab appear to be the treatment of choice for AMD patients with nonsubfoveal CNV. PMID- 20027410 TI - Comparative study of the blinking time between young adult and adult video display terminal users in indoor environment. AB - OBJECTIVE: Investigate the average blinking time in conversation and in Video Display Terminal use of young adults and adults in the presbyopic age group. METHODS: A transversal analytical study in a readily accessible sample consisting of Volkswagen do Brasil - Curitiba, Parana employees was performed. The cohort group consisted of 108 subjects divided into two age groups: Group 1, the young adult group (age range 20-39): 77 employees, mean age of 30.09 +/- 5.09; Group 2, the presbyopic adult group, (age range 40-53): 31 employees, mean age of 44.17 +/ 3. Subjects under 18 years of age, with a history of ocular disorders, contact lens wearers and computer non-users were excluded. The subjects had their faces filmed for 10 minutes in conversation and VDT reading. Student's t-test was used and the statistical significance level was 95%. RESULTS: The average time between blinks in Group 1 for conversation and VDT reading was 5.16 +/- 1.83 and 10.42 +/ 7.78 seconds, respectively; in Group 2. 4,9 +/- 1.49 and 10.46 +/- 5.54 seconds. In both age groups, the time between blinks in VDT reading situations was higher (p<0.0001). There was no statistically meaningful difference for conversation and VDT reading situations when the two studied age groups were compared (p>0.05). CONCLUSION: There was an increase in the blinking time between young adults and the presbyopic group in VDT use situations when compared with reading situations. The difference in the blinking frequency between young adults and the presbyopic group in VDT use and reading situations was not statistically significant. PMID- 20027411 TI - [Correction of ocular aberrations in custom and standard LASIK retreatments]. AB - PURPOSE: To compare the correction of ocular aberrations between custom and standard LASIK retreatment. METHODS: Prospective, randomized trial with paired eye control of 74 eyes from 37 patients who underwent LASIK retreatment. Each patient underwent retreatment using Zyoptix LASIK (Bausch & Lomb) in 1 eye and Planoscan LASIK (Bausch & Lomb) in the fellow eye. Correction of ocular aberrations was compared between custom and standard LASIK retreatments. RESULTS: At 6 months, there was a statistically significant reduction in defocus, astigmatism, coma, spherical aberration, second, third, higher-order and total aberration in Zyoptix eyes. There was a statistically significant reduction in defocus, second-order and total aberration in Planoscan eyes. CONCLUSIONS: Custom retreatment was statistically superior than standard retreatment for correction of lower and higher ocular aberrations. PMID- 20027412 TI - Pigmentary retinopathy due to Bardet-Biedl syndrome: case report and literature review. AB - Bardet-Biedl syndrome (BBS) is a rare autosomal recessive disorder with clinical and genetic heterogeneity. This syndrome was first described by Laurence and Moon in 1866 and additional cases were described by Bardet and Biedl between 1920 and 1922. The main features are obesity, polydactyly, pigmentary retinopathy, learning disabilities, various degrees of intellectual impairment, hypogonadism, and renal abnormalities. Bardet-Biedl syndrome is both phenotypically and genetically heterogeneous. Clinical diagnosis is based on the presence of 4 of the 5 cardinal features. The authors present a typical case of pigmentary retinopathy due to Bardet-Biedl syndrome and made a brief commentary about the disease's cardinal manifestations. PMID- 20027413 TI - [Posterior segment changes in microphthalmic eyes: case report]. AB - Microphthalmos is a developmental ocular disorder defined as a small eyeball. The condition can be associated with abnormalities of anterior and posterior segments. The most common anterior characteristics include corneal opacities, angle-closure and a shallow anterior chamber and cataract. The main findings of posterior segment are uveal effusion, retinal folds, abnormalities of macular capillar vascularization, absence of foveal depression and peripheral retinoschisis. Three patients with microphthalmos were assisted and their OCT features of posterior segment were analyzed. The first case had uveal effusion syndrome, choroidal and retinal detachment treated with parcial sclerectomy at the four quadrants. The other case presented with neurosensory retinal fold at fovea and papillomacular area in both eyes without involvement of retinal pigment epithelium layer and choriocapillaris. The third patient showed absence of foveal depression. The main diferencial diagnosis for this condition is foveal hypoplasia. PMID- 20027414 TI - [Intracranial tumors in patients referred for optical coherence tomography examination as glaucoma suspects: case report]. AB - Optical coherence tomography (OCT) has proved to be a very valuable tool in the assessment of patients with glaucoma. In this report, intracranial tumors were discovered in two glaucoma suspects referred for diagnostic confirmation by OCT - a clivus chordoma and a craniopharyngeoma. Optical coherence tomography findings marked asymmetrical diffuse attenuation of the peripapillary nerve fiber layer in nasal and temporal sectors - raised concerns about lesions in chiasmatic region and permitted the timely diagnosis of these intraocular tumors. PMID- 20027415 TI - [Melanosis in eyelid margin with malignancy suspect: case report]. AB - A case of a patient with an irregular pigmented lesion of the lower eyelid margin simulating malignant tumor, which was treated based on the results of impression cytology and diagnosed by histopathological study is presented. The importance of cytological technique is emphasized as an effective and safe method that avoids unnecessary and extensive procedures. PMID- 20027416 TI - [Toxic maculopathy caused by antimalarial drugs: detection using spectral domain OCT: case reports]. AB - Antimalarial drugs, such as chloroquine, are useful in the management of rheumatic diseases, but may cause a potentially blinding condition known as toxic maculopathy. This report describes the findings on standard and on high resolution spectral domain ocular coherence tomography performed in a CirrusTM HD OCTdevice in two patients with chloroquine maculopathy. In one case, a very similar aspect of retinal pigment epithelium (RPE) plaque atrophy occured in both angiographic and HD-OCT images obtained by 'en-face' (coronal) segmentation. In another patient with clinical signs of maculopathy and no angiographic abnormalities, signs of RPE atrophy could also be observed in HD-OCT scans, raising the possibility that this technique may allow the early detection of the disease. PMID- 20027417 TI - [The lacrimo-auriculo-dento-digital syndrome (LADD): case report and literature review]. AB - Levy-Hollister or lacrimo-auriculo-dento-digital (LADD) syndrome is a rare entity with autossomic dominant inheritance occuring as an isolated form or affecting many family generations. Diagnosis is based on the identification of the lacrimal drainage system abnormalities with reduction or absence of tear production and bone, teeth, salivar glands and outer ear abnormalities. A 13 year-old male patient has been followed at the Hospital Servidor Publico Estadual in Sao Paulo due to dry eye since his first year of life. Due to the occurrence of early ocular manifestations in patients with Levy-Hollister or lacrimo-auriculo-dento digital syndrome, ophthalmologists must be aware to recognize and control this syndrome. PMID- 20027418 TI - [Retinocytoma: report of five cases]. AB - Five retinocytoma cases diagnosed over 75 months are reported. The mean age at diagnosis was 193.2 months. Two cases were unilateral, one was bilateral and two exhibited retinoblastoma in the fellow eye. Clinical and ophthalmoscopic aspects, genetical implications and the need of treatment and continuous follow-up of retinocytoma patients are discussed. PMID- 20027419 TI - [Macular serpiginous choroiditis: case report]. AB - Serpiginous choroiditis is a rare ocular inflammatory disease that can lead to permanent vision loss due to macular involvement. This article reports a case of a patient with macular serpiginous choroiditis submitted to subtenon and intravitreal triamcinolone injections. The clinical aspects and differential diagnosis of this ocular disease, including fluorescein angiogram and high resolution optical coherence tomography are discussed. PMID- 20027420 TI - [Ocular leprosy: a historical approach]. AB - A sharp drop in the prevalence of leprosy occurred in the last three decades. However, the incidence has not decreased at the same rate. Three years after the World Health Organization last deadline for leprosy control, patients considered healed still need special care for their incapacities and immunopathological reactions. Medical literature reffers blindness in 4% to 11% of studied patients and more than 20% with severe visual problems due to corneal exposure, bacillary invasion and hipersensibility. These mechanisms result in a population of nearly one million blind leprosy patients even though official prevalence accounts no more than 250,000 patients worldwide. The author calls for better patients management and follow-up and urges ophthalmologists to become more aware and interested in the treatment of the ocular complications of leprosy. PMID- 20027421 TI - Molecular biology applied to the laboratory diagnosis of bacterial endophthalmitis. AB - Bacterial endophthalmitis is a serious but uncommon intraocular infection which frequently results in vision loss. Early diagnosis and appropriate therapy are associated with better visual outcome. Conventional microbiological methods are currently used for microbiological characterization of eyes with suspected endophthalmitis. However, the sensitivity of bacterial detection from aqueous and vitreous humor using microbiology techniques is poor, and time-consuming to confirm the results. The application of molecular methods enhances significantly laboratory confirmation of bacterial endophthalmitis, demanding a shorter time to draw a definitive result and thereby promoting the early initiation of a more specific therapy to limit the empirical or unnecessary use of broad-spectrum antibiotics. PCR-based techniques, including post-PCR methods such RFLP, DNA probe hybridization and DNA sequencing have been successfully used for the diagnostic elucidation of clinically suspected bacterial endophthalmitis cases, showing promising application in the routine practice of ocular microbiology laboratories. PMID- 20027423 TI - Guest Editorial: referred pain. PMID- 20027424 TI - Cytotoxicity and biocompatibility of direct and indirect pulp capping materials. AB - There are several studies about the cytotoxic effects of dental materials in contact with the pulp tissue, such as calcium hydroxide (CH), adhesive systems, resin composite and glass ionomer cements. The aim of this review article was to summarize and discuss the cytotoxicity and biocompatibility of materials used for protection of the dentin-pulp complex, some components of resin composites and adhesive systems when placed in direct or indirect contact with the pulp tissue. A large number of dental materials present cytotoxic effects when applied close or directly to the pulp, and the only material that seems to stimulate early pulp repair and dentin hard tissue barrier formation is CH. PMID- 20027425 TI - Antimicrobial activity of sodium hypochlorite associated with intracanal medication for Candida albicans and Enterococcus faecalis inoculated in root canals. AB - OBJECTIVE: The purpose of this study was to evaluate the action of sodium hypochlorite (NaOCl) associated with an intracanal medication against Candida albicans and Enterococcus faecalis inoculated in root canals. MATERIAL AND METHODS: Thirty-six human single-rooted teeth with single root canals were used. The canals were contaminated with C. albicans and E. faecalis for 21 days and were then instrumented with 1% NaOCl. The roots were divided into 3 groups (n=12) according to the intracanal medication applied: calcium hydroxide paste, 2% chlorhexidine (CHX) gel, and 2% CHX gel associated with calcium hydroxide. The following collections were made from the root canals: a) initial sample (IS): 21 days after contamination (control), b) S1: after instrumentation, c) S2: 14 days after intracanal medication placement; S3: 7 days after intracanal medication removal. The results were analyzed statistically by the Kruskal-Wallis test at 5% significance level. RESULTS AND CONCLUSIONS: Both 1% NaOCl irrigation and the intracanal medications were effective in eliminating E. faecalis and C. albicans inoculated in root canals. PMID- 20027426 TI - Protective effect of green tea on dentin erosion and abrasion. AB - OBJECTIVE: This in situ study evaluated the protective effect of green tea on dentin erosion (ERO) and erosion-abrasion (ABR). MATERIAL AND METHODS: Ten volunteers wore intraoral palatal appliances with bovine dentin specimens subjected to ERO or ERO + toothbrushing abrasion performed immediately (ERO+I ABR) or 30 min after erosion (ERO+30-min-ABR). During 2 experimental 5-day crossover phases, the volunteers rinsed with green tea or water (control, 1 min) between each erosive (5 min, cola drink) and abrasive challenge (30 s, toothbrushing), 4x/day. Dentin wear was measured by profilometry. RESULTS: The green tea reduced the dentin wear significantly for all conditions compared to control. ERO+I-ABR led to significantly higher wear than ERO, but it was not significantly different from ERO+30-min-ABR. ERO+30-min-ABR provoked significant higher wear than ERO, only for the placebo treatment. CONCLUSIONS: From the results of the present study, it may be concluded that green tea reduces the dentin wear under erosive/abrasive conditions. PMID- 20027427 TI - A survey of cross-infection control procedures: knowledge and attitudes of Turkish dentists. AB - OBJECTIVES: The objective of this study was to investigate the knowledge, attitudes and behavior of Turkish dentists in Samsun City regarding cross infection control. MATERIAL AND METHODS: A questionnaire was designed to obtain information about procedures used for the prevention of cross-infection in dental practices and determine the attitudes and perceptions of respondent dental practitioners to their procedures. The study population included all dentists in the city of Samsun, Turkey, in April 2005 (n=184). The questionnaire collected data on sociodemographic characteristics, knowledge and practice of infection control procedures, sterilization, wearing of gloves, mask, use of rubber dam, method of storing instruments and disposal methods of contaminated material, etc. Questionnaire data was entered into a computer and analyzed by SPSS statistical software. RESULTS: From the 184 dentists to whom the questionnaires were submitted, 135 participated in the study (overall response rate of 73.36%). As much as 74.10% dentists expressed concern about the risk of cross-infection from patients to themselves and their dental assistants. Forty-three percent of the participants were able to define 'cross-infection' correctly. The greatest majority of the respondents (95.60%) stated that all patients have to be considered as infectious and universal precautions must apply to all of them. The overall responses to the questionnaire showed that the dentists had moderate knowledge of infection control procedures. CONCLUSIONS: Improved compliance with recommended infection control procedures is required for all dentists evaluated in the present survey. Continuing education programs and short-time courses about cross-infection and infection control procedures are suitable to improve the knowledge of dentists. PMID- 20027428 TI - Glass transition and degree of conversion of a light-cured orthodontic composite. AB - OBJECTIVE: This study evaluated the glass transition temperature (Tg) and degree of conversion (DC) of a light-cured (Fill Magic) versus a chemically cured (Concise) orthodontic composite. MATERIAL AND METHODS: Anelastic relaxation spectroscopy was used for the first time to determine the Tg of a dental composite, while the DC was evaluated by infrared spectroscopy. The light-cured composite specimens were irradiated with a commercial LED light-curing unit using different exposure times (40, 90 and 120 s). RESULTS: Fill Magic presented lower Tg than Concise (35-84 masculineC versus 135 masculineC), but reached a higher DC. CONCLUSIONS: The results of this study suggest that Fill Magic has lower Tg than Concise due to its higher organic phase content, and that when this light cured composite is used to bond orthodontic brackets, a minimum energy density of 7.8 J/cm(2) is necessary to reach adequate conversion level and obtain satisfactory adhesion. PMID- 20027429 TI - Fracture strength of flared bovine roots restored with different intraradicular posts. AB - OBJECTIVE: The aim of this study was to evaluate the fracture strength and failure mode of flared bovine roots restored with different intraradicular posts. MATERIAL AND METHODS: Fifty bovine incisors with similar dimensions were selected and their roots were flared until 1.0 mm of dentin wall remained. Next, the roots were allocated into five groups (n=10): GI- cast metal post-and-core; GII- fiber posts plus accessory fiber posts; GIII- direct anatomic post; GIV- indirect anatomic post and GV- control (specimens without intraradicular post). A polyether impression material was used to simulate the periodontal ligament. After periodontal ligament simulation, the specimens were subjected to a compressive load at a crosshead speed of 0.5 mm/min in a servo-hydraulic testing machine (MTS 810) applied at 135 masculine to the long axis of the tooth until failure. The data (N) were subjected to ANOVA and Tukey's post-hoc test (alpha=0.05). RESULTS: GI and GIV presented higher fracture strength (p<0.05) than GII. GIII presented intermediate values without statistically significant differences (p>0.05) from GI, GII and GIV. Control specimens (GV) produced the lowest fracture strength mean values (p<0.05). Despite obtaining the highest mean value, GI presented 100% of unfavorable failures. GII presented 20% of unfavorable failures. GIII, GIV and GV presented only favorable failures. CONCLUSIONS: Although further in vitro and in vivo studies are necessary, the results of this study showed that the use of direct and indirect anatomic posts in flared roots could be an alternative to cast metal post-and-core. PMID- 20027430 TI - Effect of eugenol-based endodontic sealer on the adhesion of intraradicular posts cemented after different periods. AB - OBJECTIVE: This study evaluated in vitro the influence of an eugenol-based sealer (EndoFill) on the retention of stainless steel prefabricated posts cemented with zinc phosphate and resin-based (Panavia F) cements after different periods of root canal obturation, using the pull-out test. MATERIAL AND METHODS: Sixty upper canines were decoronated and the roots were embedded in resin blocks. The specimens were distributed into 3 groups, according to the period elapsed between canal obturation and post cementation: Group I - immediately; Group II - 72 h and Group III - 4 months. The groups were subdivided according to the type of cement used for post cementation: A - zinc phosphate and B - Panavia F. Following the experimental periods, specimens were subjected to pullout test in an Instron machine with application of tensile force at a crosshead speed of 0.5 mm/min until post dislodgement. The maximum forces required for post removal were recorded (kN) and means were subjected to statistical analysis by 2-way ANOVA and Tukey-Kramer test (alpha=0.001) RESULTS: There were statistically significant differences (p<0.01) between the posts cemented with zinc phosphate cement (0.2112 kN) and Panavia F (0.0501 kN). However, no statistically significant differences (p>0.05) were found between the three post cementation periods, regardless of the cement. CONCLUSIONS: It was concluded that the eugenol-based sealer influenced the tensile strength of the posts cemented with the resin cement, but had no influence on the time waited between root canal obturation and post space preparation/post cementation. PMID- 20027431 TI - Influence of microwave sterilization on the cutting capacity of carbide burs. AB - OBJECTIVE: This study compared the cutting capacity of carbide burs sterilized with microwaves and traditional sterilization methods. MATERIAL AND METHODS: Sixty burs were divided into 5 groups according to the sterilization methods: dry heat (G1), autoclave (G2), microwave irradiation (G3), glutaraldehyde (G4) or control - no sterilization (G5). The burs were used to cut glass plates in a cutting machine set for twelve 2.5-min periods and, after each period, they were sterilized (except G5) following the protocol established for each group. The cutting capacity of the burs was determined by a weight-loss method. Data were analyzed statistically by Kruskal-Wallis and Dunn's test. RESULTS: The means of the cutting amount performed by each group after the 12 periods were G1 = 0.2167 +/- 0.0627 g; G2 = 0.2077 +/- 0.0231 g; G3 = 0.1980 +/- 0.0326 g; G4 = 0.1203 +/- 0.0459 g; G5 = 0.2642 +/- 0.0359 g. There were statistically significant differences among the groups (p<0.05); only dry heat sterilization was similar to the control. CONCLUSION: Sterilization by dry heat was the method that least affected the cutting capacity of the carbide burs and microwave sterilization was not better than traditional sterilization methods. PMID- 20027432 TI - Assessment of canal walls after biomechanical preparation of root canals instrumented with Protaper Universal rotary system. AB - OBJECTIVE: The aim of this study was to examine the instrumented walls of root canals prepared with the ProTaper Universal rotary system. MATERIAL AND METHODS: Twenty mesiobuccal canals of human first mandibular molars were divided into 2 groups of 10 specimens each and embedded in a muffle system. The root canals were transversely sectioned 3 mm short of the apex before preparation and remounted in their molds. All root canals were prepared with ProTaper Universal rotary system or with Nitiflex files. The pre and postoperative images of the apical thirds viewed with a stereoscopic magnifier (X45) were captured digitally for further analysis. Data were analyzed statistically by Fisher's exact test and Chi-square test at 5% significance level. RESULTS: The differences observed between the instrumented and the noninstrumented walls were not statistically significant (p<0.05). CONCLUSIONS: The Nitiflex files and the ProTaper Universal rotary system failed to instrument all the root canal walls. PMID- 20027433 TI - CO(2), Er: YAG and Nd:YAG lasers in endodontic surgery. AB - OBJECTIVES: CO(2), Er:YAG and Nd:YAG lasers have been used in endodontic surgery. This in vitro study evaluated 1% Rhodamine B dye penetration using computer assisted morphometry (ImageTool Software) of 108 endodontically treated human permanent canines. MATERIAL AND METHODS: Teeth were divided into 9 groups according to the technique used: A: 90-degree apicoectomy with bur, root-end cavity preparation with ultrasound and filled with MTA; B: 90-degree apicoectomy with bur, root-end cavity prepared with ultrasound and filled with MTA, and treatment of apical surface with CO(2) laser (1 W, CW/CW); C: 90-degree apicoectomy with bur, and treatment of apical surface with Nd:YAG laser (150 mJ, 10 Hz); D: 90-degree apicoectomy with bur, and treatment of apical surface with CO(2) laser,(1 W, CW/CW); E: apicoectomy with Er:YAG laser (400 mJ, 10 Hz), root end cavity prepared with ultrasound and filled with MTA; F: apicoectomy with Er:YAG laser (400 mJ, 10 Hz) and treatment of apical surface with Nd:YAG laser (150 mJ, 10Hz); G: apicoectomy with CO(2) laser (5W, CW/SP), root-end cavity prepared with ultrasound and filled with MTA; H: irradiation of apical end with CO(2) laser (1 W, CW/CW); I: irradiation of apical end with Nd:YAG laser (150 mJ, 10 Hz). RESULTS: Dye penetration was found in all specimens at different rates, the lowest penetration occurring in groups C (16.20%), B (17.24%) and F (17.84%). CONCLUSIONS: Groups B, C and F represent the best technical sequences to perform endodontic surgery. PMID- 20027434 TI - Effect of relining on fiber post retention to root canal. AB - One of the clinically relevant problems dentists face when restoring endodontically treated teeth is the mismatch between fiber post and post space diameters, which results in an excessively thick resin cement layer. Fiber post relining appears as a solution for this problem. OBJECTIVES: The aim of this study was to evaluate the effect of fiber post relining with composite resin on push-out bond strength. MATERIAL AND METHODS: Twenty bovine incisors were selected to assess post retention. The crowns were removed below the cementoenamel junction and the root canals were treated endodontically and flared with diamond burs. The roots were allocated into two groups (n=10): G1: fiber posts without relining and G2: fiber posts relined with composite resin. The posts were cemented with a dual-cured resin cement and the specimens were sectioned transversally. Three 1.5-mm thick slabs were obtained per root and identified as cervical, medium and apical thirds. The push-out test was performed at a crosshead speed of 0.5 mm/min until post dislodgement occurred. The failure mode of fractured specimens was analyzed under scanning electron microscopy. Data were analyzed by split-plot ANOVA and post-hoc Tukey's test at a pre-set alpha of 0.05. RESULTS: Relined fiber posts presented higher retention values than non relined post in all thirds. No statistically significant differences (p>0.05) were found among thirds for relined posts. All failures occurred at the interface between resin cement and root dentin. CONCLUSIONS: Relining with composite resin seems to be an effective method to improve the retention of fiber posts to flared root canals. PMID- 20027435 TI - Effect of immediate and delayed post space preparation on the apical seal of root canals obturated with different sealers and techniques. AB - During mechanical preparation of the post space, the root canal filling may be twisted or vibrated, depending on several factors associated with the preparation technique and quality of filling. OBJECTIVE: The purpose of this study was to investigate the effect of immediate and delayed post space preparation on the integrity of the apical seal. MATERIAL AND METHODS: Sixty-four extracted human incisors were biomechanically prepared using the step-back technique. Sixty roots were randomly assigned to 6 experimental groups of 10 teeth each and the remaining 4 roots served as positive and negative controls (n=2). The root canals in the different groups were obturated with cold lateral and warm vertical condensation of gutta-percha and one of two sealers (Sealapex and Diaket). Post space was prepared either individually or simultaneously. An insulated copper wire was cut into 10-cm-long pieces. In each canal, one piece was inserted to maintain contact with gutta-percha and extended to the outside as one of two working electrodes. A stainless steel wire with the same dimensions of those of the copper wire, used as the other working electrode, was immersed into the background electrolyte from the center of the bottle. The electrical current between standard and experimental electrodes in canals was measured over a period of 10 days applying a conductivity meter. The Kruskal-Wallis test (p=0.05) determined whether there was a significant difference in microleakage among the groups and the Mann-Whitney U test (p=0.01) was used for multiple comparison grouping variables. RESULTS: The results suggest that only the differences between the root canal filling techniques were statistically significant (p<0.01). There were no statistically significant difference in the sealers and the times of filling removal for post space preparation (p>0.01). CONCLUSION: The quality of the root canal filling is important for the integrity of the apical seal. PMID- 20027436 TI - Micromorphology and microhardness of enamel after treatment with home-use bleaching agents containing 10% carbamide peroxide and 7.5% hydrogen peroxide. AB - OBJECTIVE: The purpose of this study was to assess the effect of home-use bleaching agents containing 10% carbamide peroxide and 7.5% hydrogen peroxide on enamel microhardness and surface micromorphology. MATERIAL AND METHODS: Enamel slabs (n=10) received the bleaching agents for 1 h/day and remained in artificial saliva solution for 23 h/day, during a total period of 21 days. Control group was composed of enamel slabs that were not subjected to treatment with the agents and were maintained in artificial saliva solution. Microhardness tests were performed before treatment application, 21 days of treatment and 14 days after the end of treatment. Scanning electron microscopy analyses were performed after 14 days after the end of bleaching treatment by 3 calibrated observers who attributed scores. RESULTS: The Tukey's test (alpha=0.05) showed no significant differences in microhardness values among bleaching agents, at 21 days of treatment and a significant increase in microhardness for different agents after 14 days from the end of treatment. Fisher's exact test showed differences in micromorphology of enamel between control and experimental groups (p=0.0342). CONCLUSIONS: Bleaching agents containing 10% carbamide peroxide and 7.5% hydrogen peroxide may change surface micromorphology of enamel, although no changes in microhardness were observed. PMID- 20027437 TI - Influence of post-cure treatments on hardness and marginal adaptation of composite resin inlay restorations: an in vitro study. AB - OBJECTIVES: The purpose of this study was to evaluate the Vickers hardness number (VHN) and the in vitro marginal adaptation of inlay restorations of three hybrid composite resins (Filtek Z250, Opallis and Esthet-X) subjected to two post-cure treatments. MATERIAL AND METHODS: For the microhardness test, three different groups were prepared in accordance with the post-cure treatments: control group (only light cure for 40 s), autoclave group (light cure for 40 s + autoclave for 15 min at 130 masculineC); and microwave group (light cure for 40 s + microwave for 3 min at 450 W). To assess the marginal adaptation, the composite resin was inserted incrementally into a mesial-occlusal-distal cavity brass mold and each increment light-cured for 40 s. A previous reading in micrometers was taken at the cervical wall, using a stereomicroscope magnifying glass equipped with a digital video camera and image-analysis software. Subsequently, the specimens were subjected to the post-cure treatments (autoclave and microwave) and a reading was taken again at the cervical wall. Data were compared using ANOVA for the hardness test, split-plot ANOVA for the adaptation assessment and Tukey's test for multiple comparisons. A significance level of 5% was adopted for all analyses. RESULTS: The post-cure treatments increased the hardness of conventional composites (p<0.001) and the gap values of inlay restorations (p<0.01). Filtek Z250 showed higher hardness (p<0.001) and lower gap values than Opallis and Esthet-X (p<0.05). Gap values did not exceed 90 microm for any of the experimental conditions. CONCLUSION: The post-cure treatments increased the VHN and the gap values on the cervical floor of composite resin inlays. Moreover, Filtek Z250 showed the best results, with higher hardness and lower gap values. PMID- 20027438 TI - Alport syndrome: significance of gingival biopsy in the initial diagnosis and periodontal evaluation after renal transplantation. AB - Alport Syndrome (AS) is an important hereditary disorder affecting the glomerular basement membrane. Diagnosis of AS is based on the presence of hematuric nephropathy, renal failure, hearing loss, ocular abnormalities and changes in the glomerular basement membrane of the lamina densa. The aims of this case report were to show the changes in the gingival tissues in a patient with AS under therapy with cyclosporin-A after renal transplantation and to discuss the possible role of type IV collagen in gingival basal lamina as an alternative approach for the diagnosis of AS. A 20-year-old male patient with AS underwent periodontal therapy including a series of gingivectomy surgeries. Gingival samples obtained during the second surgery were examined histopathologically and by transmission electron microscopy for further pathological examination. Gingivectomy procedures have been performed every 6 months over the last 4 years. The excessive and fibrous gingival enlargements resulted in migration of the anterior teeth, but no alveolar bone loss occurred. This is the first report to demonstrate the possible changes in the gingival tissues caused by AS. It is suggested that gingival biopsy can be an initial diagnostic tool instead of renal or skin biopsies. Proper dental and periodontal care and regular visits to the dentist could provide limited gingival hyperplasia to patients with AS. PMID- 20027440 TI - Differences in responses to the Oral Health Impact Profile (OHIP14) used as a questionnaire or in an interview. AB - The objective of this study was to compare the completion rates and performance of the Brazilian version of the Oral Health Impact Profile (OHIP14) when applied as an interview or in its original self-reported form. A convenience sample of 74 adult patients was selected in a Dental Clinic (University of Araras, Brazil). One examiner administered the instrument in both formats to participants with an interval of 2 weeks between each administration. Data about dental health condition and socioeconomic status were collected and associated with total OHIP14 scores in both formats using linear regression analyses. No differences were found in the total scores and in each subscale of the OHIP14 according to the form of administration. Higher values of completion were found in the interview format. More severe impacts were recorded in the interview format than in the questionnaire format. Higher values of total OHIP-14 scores in both formats were related to the presence of dental caries. Total OHIP14 scores were not influenced by the method of administration. However, the use of the OHIP14 in the questionnaire format may result in lower completion rates and loss of data. PMID- 20027441 TI - Relationship of body mass index with periodontal health status of green marble mine laborers in Kesariyaji, India. AB - It is evident from literature that an increased body mass index (BMI) may be a potential risk factor for periodontitis. Association between BMI and periodontitis has been ascribed to unhealthy dietary patterns with insufficient micronutrients and excess sugar and fat content. The present study population has been plagued by unhealthy nutritional practices, hence the present study intended to assess the relation between BMI and periodontal status among green marble mine laborers of Kesariyaji, in the Udaipur district of Rajasthan, India. The study sample comprised of 513 subjects aged 18-54 years, drawn using the stratified cluster sampling procedure. BMI was calculated as the ratio of the subject's body weight (in kg) to the square of their height (in meters). Periodontal status was recorded using the Community Periodontal Index (CPI). Binary multiple logistic regression analysis was executed to assess the relation between body mass index and periodontitis. The dependent variable for logistic regression analysis was categorized into control group (scores 0 - 2 of the CPI) and periodontitis group (scores 3 and 4 of the CPI). The overall prevalence of periodontal disease was 98.2%. Caries status and mean number of teeth present deteriorated with the poor periodontal status. Subjects had an increased risk of periodontitis by 57% for each 1kg/m(2) increase in the body mass index, which means that a higher body mass index could be a potential risk factor for periodontitis among the adults aged 18 to 54 years. In conclusion, evaluation of the body mass index could be used in periodontal risk assessment. PMID- 20027442 TI - Oral hygiene and periodontal status among Terapanthi Svetambar Jain monks in India. AB - The main objective of the study was to determine the oral hygiene levels and periodontal status among Jain monks attending a Chaturmass in Udaipur, India. To date, no study has been conducted on Jain monks. The study comprises of 180 subjects and the overall response rate was 76% among them. Oral hygiene status was assessed by the Simplified Oral Hygiene Index (OHI-S) of Greene, Vermillion14 (1964), and periodontal status was assessed by the Community Periodontal Index. Additional information was collected regarding food habits, education level and oral hygiene habits. Analysis of variance (ANOVA), Chi Square Test and Step-wise multiple linear regression analysis were carried out using SPSS Software (11.0). The results showed that the oral hygiene status of Jain monks was poor and only 5.6% of the subjects had good oral hygiene. Overall periodontal disease prevalence was 100% with bleeding and shallow pocket contributing a major part (72.8%) among all the age groups (p < 0.001). Multiple linear regression analysis revealed that oral hygiene habits, caloric intake and education level explained a variance of 11.7% for the Oral hygiene index collectively. The findings confirmed that Jain monks have poor oral hygiene and an increased prevalence of periodontal disease compared to that of the similarly aged general population because, as a part of their religion, many Jain individuals avoid brushing their teeth especially during fasting, keeping in mind not to harm the microorganisms present in the mouth. PMID- 20027443 TI - Qualitative assessment of the dental health services provided at a dental school in Kerman, Iran. AB - Increasing the quality of the services provided in a Dental School can raise the satisfaction level of patients and consequently increase the level of their oral health. This study was conducted to evaluate the quality of dental care and services provided to patients referred to a Dental School in Kerman, Iran. In this qualitative study, face-to-face, in-depth interviews were conducted with 41 participants [25 patients (P), 5 nurses (N), 6 dental academic staff (AS), and 5 dental students (S)]. Then, the interviews were transcribed and analyzed, using content analysis of data. Data analysis in qualitative research involves breaking down the data and searching for codes and categories that are then reassembled to form themes. Both positive and negative themes emerged. Positive themes included: good infection control, service accessibility, patient appointments and visits were not assigned on merit, precise examinations, and comprehensive treatment plans. Negative themes included: long wait time, lack of options to pass waiting time, such as newspapers and television, an insufficient number of nurses, and not enough professors for supervision. In addition, the results of this study show that the patients and dental staff have high expectations in relation to dental services, and that implementation of these expectations would increase the overall satisfaction with and the quality of the level of services. Finally, some recommendations for improving services in the Kerman Dental School were given to the managing team of the Dental School. PMID- 20027444 TI - Effect of sodium bicarbonate on Candida albicans adherence to thermally activated acrylic resin. AB - The purpose of this study was to evaluate the effect of 5% sodium bicarbonate on the adherence of Candida albicans to thermally activated acrylic resin. Fifty 4 mm(2) specimens of acrylic resin were obtained using a metallic matrix. The specimens received chemical polishing, were sterilized and then immersed in Sabouraud broth, inoculated with Candida albicans standardized suspension. After 24 hours of incubation at 37 degrees Celsius, the specimens were divided into four groups according to the substance used for disinfection (5% sodium bicarbonate, 0.12% digluconate chlorhexidine, vinegar and Corega Tabs). A control group was included, in which distilled water was used. The adhered microorganisms were dispersed, diluted and plated onto culture media to determine the number of colony-forming units (cfu/mL). The results were analyzed through the Mann-Whitney statistical test at the 5% level of significance. Only 0.12% digluconate chlorhexidine and 5% sodium bicarbonate presented a statistically significant difference (p = 0.0010 and p = 0.0156, respectively) compared to the control group, decreasing the number of cfu/mL. However, when the different disinfecting solutions were compared with each other, only 0.12% digluconate chlorhexidine presented a statistically significant difference in the reduction of cfu/mL. It was concluded that although 0.12% digluconate chlorhexidine was more effective in the reduction of Candida albicans adherence values to thermally activated acrylic resin, 5% sodium bicarbonate also proved to be a viable alternative. PMID- 20027445 TI - Color alteration of the paint used for iris painting in ocular prostheses. AB - The purpose of this study was to assess color alteration of the paints used for iris painting in artificial eyes. Five disks of heat cured acrylic resin were confectioned by microwave energy for each paint analyzed, in a total of 40 specimens. Each specimen consisted of a colorless acrylic resin disk and another of equal size, of scleral white colored acrylic resin, with the painting interposed between the two disks. The specimens were submitted to an accelerated aging process in a chamber under ultraviolet radiation for 1,008 hours. To assess color variation, a reflective spectrophotometer was used. The results were statistically analyzed by ANOVA and the Tukey test (p < 0.05). All the paints underwent chromatic alteration. The oil paint presented the highest resistance to accelerated aging. PMID- 20027446 TI - Immunohistochemical study of androgen, estrogen and progesterone receptors in salivary gland tumors. AB - The aim of this work was to study the immunohistochemical expression of androgen receptor, estrogen receptor and progesterone receptor in pleomorphic adenomas, Warthin's tumors, mucoepidermoid carcinomas and adenoid cystic carcinomas of salivary glands. A total of 41 pleomorphic adenomas, 30 Warthin's tumors, 30 mucoepidermoid carcinomas and 30 adenoid cystic carcinomas were analyzed, and the immunohistochemical expression of these hormone receptors were assessed. It was observed that all cases were negative for estrogen and progesterone receptors. Androgen receptor was positive in 2 cases each of pleomorphic adenoma, mucoepidermoid carcinoma and adenoid cystic carcinoma. In conclusion, the results do not support a role of estrogen and progesterone in the tumorigenesis of pleomorphic adenomas, Warthin's tumors, mucoepidermoid carcinomas and adenoid cystic carcinomas. However, androgen receptors can play a role in a small set of salivary gland tumors, and this would deserve further studies. PMID- 20027447 TI - An in vitro comparison of nickel and chromium release from brackets. AB - This study aimed at comparing amounts of nickel (Ni) and chromium (Cr) released from brackets from different manufacturers in simulated oral environments. 280 brackets were equally divided into 7 groups according to manufacturer. 6 groups of brackets were stainless steel, and 1 group of brackets was made of a cobalt chromium alloy with low Ni content (0.5%). International standard ISO 10271/2001 was applied to provide test methods. Each bracket was immersed in 0.5 ml of synthetic saliva (SS) or artificial plaque fluid (PF) over a period of 28 days at 37 degrees Celsius. Solutions were replaced every 7 days, and were analyzed by spectrometry. The Kruskal-Wallis test was applied. Amounts of Ni release in SS (microg L(-1) per week) varied between groups from 'bellow detection limits' to 694, and from 49 to 5,948.5 in PF. The group of brackets made of cobalt-chromium alloy, with the least nickel content, did not release the least amounts of Ni. Amounts of Cr detected in SS and in PF (microg L(-1) per week) were from 1 to 10.4 and from 50.5 to 8,225, respectively. It was therefore concluded that brackets from different manufacturers present different corrosion behavior. Further studies are necessary to determine clinical implications of the findings. PMID- 20027448 TI - Comparison of the shear bond strengths of conventional mesh bases and sandblasted orthodontic bracket bases. AB - This study aimed to compare in vitro the shear bond strength between metallic brackets (Abzil) with conventional mesh bases and metallic brackets with bases industrially sandblasted with aluminum oxide using three adhesive systems, in order to assess the influence of sandblasting on adhesiveness and to compare 3 different bonding systems. Two hundred and forty bovine incisors were used and randomly divided into 6 groups (40 teeth in each group), according to the bracket base and to the bonding system. The brackets were direct-bonded in bovine teeth with 3 adhesive systems: System A - conventional Transbond XT (3M - Unitek); System B - Transbond Plus Self Etching Primer + Transbond XT (3M - Unitek) and System C - Fuji ORTHO LC resin-reinforced glass ionomer cement in capsules (GC Corp.). Shear bond strength tests were performed 24 hours after bonding, in a DL 3000 universal testing machine (EMIC), using a load cell of 200 kgf and a speed of 1 mm/min. The results were submitted to statistical analysis and showed no significant difference between conventional and sandblasted bracket bases. However, comparison between the bonding systems presented significantly different results. System A (14.92 MPa) and system C (13.24 MPa) presented statistically greater shear bond strength when compared to system B (10.66 MPa). There was no statistically significant difference between system A and system C. PMID- 20027449 TI - Electromyographic evaluation of the upper lip according to the breathing mode: a longitudinal study. AB - The present study aimed at analyzing and comparing longitudinally the EMG (electromyographic activity) of the superior orbicularis oris muscle according to the breathing mode. The sample, 38 adolescents with Angle Class II Division 1 malocclusion with predominantly nose (PNB) or mouth (PMB) breathing, was evaluated at two different periods, with a two-year interval between them. For that purpose, a 16channel electromyography machine was employed, which was properly calibrated in a PC equipped with an analogue-digital converter, with utilization of surface, passive and bipolar electrodes. The RMS data (root mean square) were collected at rest and in 12 movements and normalized according to time and amplitude, by the peak value of EMG, in order to allow comparisons between subjects and between periods. Comparison of the muscle function of PNB and PMB subjects at period 1 (P1), period 2 (P2) and the variation between periods (Delta) did not reveal statistically significant differences between groups (p < 0.05). However, longitudinal evaluation of the muscle function in PNB and PMB subjects demonstrated different evolutions in the percentage of required EMG for accomplishment of the movements investigated. It was possible to conclude that there are differences in the percentage of electric activity of the upper lip with the growth of the subjects according to the breathing mode. PMID- 20027450 TI - Rhinometric evaluation of nasal cavity geometry and its relation to the upper arch transverse distance. AB - The objective of this study was to evaluate children's respiratory patterns in the mixed dentition, by means of acoustic rhinometry, and its relation to the upper arch width development. Fifty patients were examined, 25 females and 25 males with mean age of eight years and seven months. All of them were submitted to acoustic rhinometry and upper and lower arch impressions to obtain plaster models. The upper arch analysis was accomplished by measuring the interdental transverse distance of the upper teeth, deciduous canines (measurement 1), deciduous first molars (measurement 2), deciduous second molars (measurement 3) and the first molars (measurement 4). The results showed that an increased left nasal cavity area in females means an increased interdental distance of the deciduous first molars and deciduous second molars and an increased interdental distance of the deciduous canines, deciduous first and second molars in males. It was concluded that there is a correlation between the nasal cavity area and the upper arch transverse distance in the anterior and mid maxillary regions for both genders. PMID- 20027451 TI - Effect of breast- and bottle-feeding duration on the age of pacifier use persistence. AB - This study evaluated the effect of breast- and bottle-feeding duration on the age of pacifier use persistence. Questionnaires (n = 723) with information on nutritive and nonnutritive sucking habits of children aged 3-6 years were assessed. The sample was divided according to breastfeeding duration: G1 - non breastfed, G2 - up to 3 months, G3 - discontinued between 4 and 6 months, G4 - discontinued between 7 and 12 months, and G5 - longer than 12 months. The children were also assigned to 4 groups by age of pacifier use persistence, as well as by age of bottle-feeding persistence: no habits, up to 2 years, 3-4 years and 5-6 years. Associations between nutritive sucking habits and pacifier use were analyzed using logistic regression. The larger breastfeeding groups were G2 (37.9%) and G4 (19.4%). Many children discontinued pacifier use and bottle feeding at 3-4 years of age (24.9% and 40.1%, respectively). Chances of non breastfed children (G1) with prolonged pacifier-sucking habits, in the three age ranges, were progressively higher in comparison with group G4 (OR: 4.0-7.5, p < 0.01). When comparing bottle-fed with non bottle-fed children, the age range at which bottle-feeding had been discontinued was significantly associated with that of pacifier use cessation: up to 2 years (OR = 6.2), 3-4 years (OR = 7.6) and 5-6 years (OR = 27.0), p < 0.01. It may be suggested that breastfeeding duration has an inversely proportional effect on the age of pacifier use persistence. Bottle fed children who use pacifiers tend to discontinue these habits at the same period. PMID- 20027452 TI - Microhardness of glass ionomer cements indicated for the ART technique according to surface protection treatment and storage time. AB - The aim of this study was to assess the microhardness of 5 glass ionomer cements (GIC) - Vidrion R (V, SS White), Fuji IX (F, GC Corp.), Magic Glass ART (MG, Vigodent), Maxxion R (MR, FGM) and ChemFlex (CF, Dentsply) - in the presence or absence of a surface protection treatment, and after different storage periods. For each GIC, 36 test specimens were made, divided into 3 groups according to the surface protection treatment applied - no protection, varnish or nail varnish. The specimens were stored in distilled water for 24 h, 7 and 30 days and the microhardness tests were performed at these times. The data obtained were submitted to the ANOVA for repeated measures and Tukey tests (alpha = 5%). The results revealed that the mean microhardness values of the GICs were, in decreasing order, as follows: F > CF = MR > MG > V; that surface protection was significant for MR, at 24 h, without protection (64.2 + or - 3.6a), protected with GIC varnish (59.6 + or - 3.4b) and protected with nail varnish (62.7 + or - 2.8ab); for F, at 7 days, without protection (97.8 + or - 3.7ab), protected with varnish (95.9 + or - 3.2b) and protected with nail varnish (100.8 + or - 3.4a); and at 30 days, for F, without protection (98.8 + or - 2.6b), protected with varnish (103.3 + or - 4.4a) and protected with nail varnish (101 + or - 4.1ab) and, for V, without protection (46 + or - 1.3b), protected with varnish (49.6 + or - 1.7ab) and protected with nail varnish (51.1 + or - 2.6a). The increase in storage time produced an increase in microhardness. It was concluded that the different GICs, surface protection treatments and storage times could alter the microhardness values. PMID- 20027453 TI - Therapeutic effect of two fluoride varnishes on white spot lesions: a randomized clinical trial. AB - The aim of this randomized clinical trial study was to evaluate the therapeutic effect of two varnish formulations (G1 = 5% NaF, G2 = 6% NaF + 6% CaF(2)) on the remineralization of white spot lesions (WSL). The sample was composed of 15 (7- to 12-year-old) children with 45 active WSL in anterior permanent teeth. The children were randomly divided into two groups providing 22 lesions for G1 and 23 for G2. The children were submitted to weekly varnish applications 4 times. The WSL were evaluated twice: baseline and on week 4. Maximum lesion dimensions (mesiodistal and incisogingival) were measured in millimeters and classified in four grades of size. WSL were also assessed regarding lesion activity by one calibrated examiner. The Pearson chi-square and Fisher's exact tests were used (P < 0.01). WSL reductions were observed in both varnish groups (Chi-square = 0.15, d.f. = 1, P = 0.90), and with similar magnitude (in mm): 1.19 and 1.29 for G1 and G2, respectively. Thirty-six WSL (15 in G1 and 21 in G2) were classified as inactive on week 4, reaching an overall value of 80%. No difference was observed between G1 and G2 regarding activity scores (Fisher's exact test, p > 0.01). It was concluded that after 4 applications the two varnish formulations tested produced similar clinical effects, indicating the reduction and the control of carious activity in most WSL. PMID- 20027454 TI - Influence of IL-1 gene polymorphism on the periodontal microbiota of HIV-infected Brazilian individuals. AB - This study investigated the association of IL-1A (+4845) and IL-1B (+3954) gene polymorphism with the subgingival microbiota and periodontal status of HIV infected Brazilian individuals on highly active antiretroviral therapy (HAART). One hundred and five subjects were included in the study, distributed into 2 HIV groups [29 chronic periodontitis (CP+) and 30 periodontally healthy (H+)]; and 2 non-HIV groups (29 CP- and 17 H- patients). IL-1A and B were genotyped by PCR and restriction enzyme digestion. Thirty-three bacterial species were detected by checkerboard. Overall, we observed a prevalence of the allele 2 in the IL1-A and IL-1B polymorphism at 30.5% and 25.7%, respectively. Only 11.4% of all patients were composite genotype-positive, and 75% of those were HIV-infected. No significant associations between polymorphism of the IL-1 gene and periodontitis or HIV infection were observed. Likewise, no significant differences in the frequency and counts of any bacterial species were found between individuals with and without allele 2 (IL-1A or IL-1B). The data indicated that the IL-1 gene polymorphism is neither associated with periodontal destruction nor with high levels of subgingival species, including putative periodontal pathogens in HIV Brazilian individuals on HAART. PMID- 20027455 TI - Low caloric value of ethanol itself increases alveolar bone loss in ligature induced periodontitis in male rats. AB - This study aimed at morphometrically evaluating the influence of variable caloric values of ethanol consumption on alveolar bone loss in periodontitis in male rats. Thirty-six male rats were randomized into four groups of nine rats each, as follows: Test group A (low) - rats were fed an ethanol-containing liquid diet (ethanol representing 22% of total caloric value); Control group A -rats were fed a pair-fed control diet (ethanol replaced by isocaloric amounts of carbohydrate); Test group B (high) -rats were fed an ethanol-containing liquid diet (ethanol representing 36% of total caloric value); Control group B -rats were fed a pair fed control diet for Test B. Following anesthesia, cotton ligatures were placed around the cervix of the right upper second molar. At eight weeks, the maxillary bones were removed and alveolar bone loss was analyzed by measuring the distance between the cementoenamel junction and the alveolar bone crest at buccal and palatal sites of the upper second molar. The unligated groups showed no significant differences between the bone loss values observed for the low and high caloric values of ethanol (p > 0.05). In the ligated groups, the rats receiving low caloric values of ethanol showed significantly greater bone loss compared to the isocaloric rats (p < 0.05); however, the rats receiving high caloric values of ethanol showed no significant differences compared to the controls. Analysis of the results demonstrated that, in male rats, ethanol itself affected ligature-induced bone loss when representing a low value in the total caloric value. PMID- 20027456 TI - The role of polymerization cycle and post-pressing time on tooth movement in complete dentures. AB - This study analyzed the influence of polymerization cycle and post-pressing time on tooth movement in complete dentures. Forty maxillary complete dentures were fabricated and randomly assigned to 8 groups (n = 5); the polymerization cycle (conventional long cycle in water bath, fast cycle in boiling water, and by microwave energy) and the post-pressing time (immediate and 6 hours) were varied. Metal reference pins were placed on the incisal border of the central incisors (RI and LI), on the buccal cusp of the first premolars (RP and LP), and on the mesiobuccal cusp of the second molars (RM and LM). Two transverse and 2 anteroposterior distances were measured with a linear optical microscope (Olympus Optical Co., Tokyo, Japan) with an accuracy of .0005 mm, before and after processing the complete dentures. The data collected were submitted to analysis of variance (ANOVA) and the Tukey Test at a significance level of 5%. When Classico, a conventional heat-polymerizable acrylic resin, was polymerized by microwave energy, tooth movement was statistically significant for 2 distances, with the highest value being observed for the 6-hour post-pressing time. Onda Cryl microwave acrylic resin processed by microwave energy presented the lowest tooth movement for 2 distances, considering the 6-hour post-pressing time. QC-20 acrylic resin presented no statistically significant change considering both post pressing times in each distance evaluated. Tooth movement showed an asymmetric behavior, so the mastering of the processing method is essential to the precise execution of all laboratory steps, irrespective of the polymerization cycle and post-pressing time used. PMID- 20027457 TI - Effect of different light-curing devices and aging procedures on composite knoop microhardness. AB - The aim of this study was to evaluate the effect of light-curing devices (Halogen/HAL, Light Emitting Diodes/LED, Argon Laser/LAS and Plasma Arc/PAC) and aging procedures (Mechanical Cycling/MC, Thermal Cycling/TC, Storage/S, MC+TC and MC+TC+S) on the micro-hardness of bottom/B and top/T surfaces of 2-mm-high composite resin cylinders. The Knoop microhardness test (25 g, 20 s) on both B and T was performed before and after each aging procedure. For B and T, before aging procedures, PAC showed reduced polymerization effectiveness when compared with HAL. In the T, after TC, PAC and LAS had also showed reduced polymerization effectiveness when compared to HAL and LED. For all light-curing devices, MC+TC+S and S affected the Knoop microhardness values. In the B, no difference could be observed among the aging procedures for PAC. From all light-curing units, PAC may have rendered composites of reduced quality and the storage aging procedures were the most harmful to the polymer hardness. PMID- 20027458 TI - Proven and putative vectors of American cutaneous leishmaniasis in Brazil: aspects of their biology and vectorial competence. AB - The aim of the present review is to give relevant information on aspects of the biology and ecology, including the vectorial competence of Lutzomyia sand fly species suggested as vectors of American cutaneous leishmaniasis in Brazil. The disease, due to Leishmania (Viannia) braziliensis, has been registered in most municipalities in all the Brazilian states and its transmission is associated with more than one sand fly species in each geographical region. A variety of Leishmania species can be found in the Amazon basin, where different epidemiological chains have been detected with the participation of different phlebotomine vectors. Finally, a discussion is presented on some sand fly species found naturally infected by Leishmania, but for which there is as yet no evidence regarding their epidemiological importance. PMID- 20027459 TI - Species structure of sand fly (Diptera: Psychodidae) fauna in the Brazilian western Amazon. AB - We surveyed areas of the state of Rondonia in western Amazon for phlebotomine, which are potential vectors of leishmaniasis. A total of 5,998 specimens were captured, resulting in the identification of 48 species within the Lutzomyia (99.98%) and Brumptomyia (0.02%) genera. The predominant species was Lutzomyia davisi, followed by Lutzomyia umbratilis, Lutzomyia llanosmartinsi, Lutzomyia c. carrerai, Lutzomyia dendrophyla, Lutzomyia nevesi and Lutzomyia whitmani. All sand flies identified as vectors for cutaneous leishmaniasis in Brazil, i.e., Lu. davisi, Lu. umbratilis, Lu. c. carrerai and Lu. whitmani, were found in the surveyed areas. PMID- 20027460 TI - Seroprevalence of HBV, HCV and HIV co-infection in selected individuals from state of Sao Paulo, Brazil. AB - Few studies are available on hepatitis B virus (HBV), hepatitis C virus (HCV) and human immunodeficiency virus (HIV) co-infection in populations living in small and medium-sized Brazilian cities. We evaluated the seroprevalence of these viruses in selected individuals from a clinic of infectology, who were referred to the University Regional Hospital of the West Region of state of Sao Paulo, Brazil. Among a total of 7,021 individuals seen in the clinic following receipt of preliminary ELISA results or having the suggested clinical signs of viral hepatitis or HIV, 1,228 were systematically screened. Isolated or associated HBsAg, HCV and HIV antibodies were found in 44.9% of the subjects. Anti-HIV antibodies were found in 24.7% of the patients, 20.3% of whom had an HIV monoinfection and 4.4% of whom were co-infected with hepatitis viruses (HCV: 4%; HBV: 0.4%). Anti-HCV antibodies were found in 14% of the patients and 5.9% had anti-HBsAg antibodies. HCV infection affected males more than females (p < 0.05) and individuals > 50-years old had an increased prevalence of anti-HCV compared to HIV (p = 0.0001) or HBV (p = 0.0063). HCV-RNA was detected in 73.5% of the samples with a predominance of genotype 1 (72.5%). A significant percentage (44.9%) of the selected individuals was positive for antibodies against HBV, HCV and/or HIV; these patients would otherwise have remained undiagnosed. PMID- 20027461 TI - Absonifibula estuarina sp. n. (Monogenea: Diclidophoridae) parasite of juvenile Cynoscion guatucupa (Osteichthyes) from southwestern Atlantic Ocean. AB - Absonifibula estuarina sp. n. (Diclidophoridae, Absonifibulinae), is described from the gills of juvenile striped weakfish, Cynoscion guatucupa (Cuvier), from the southwestern Atlantic, Argentinean coast. This marine fish migrates to estuarine areas to spawn where exclusively juveniles are found parasitized; adult fish in marine water were never found to be parasitized by this monogenean. A. estuarina sp. n. is characterized mainly by the pedunculate clamps dissimilar in size, the shape of anterior jaw with sclerite 'a' attached to a sub-trapezoidal lamellate extension and fused to sclerites 'c' and 'd'. It differs from Absonifibula bychowskyi Lawler & Overstreet, 1976, the only known species of the genus, in the shape and arrangement of the genital corona, which is armed with six similar hooks disposed in circle and the sub-trapezoidal shape of lamellate extension ('b'). The restriction to juvenile sciaenids is a shared feature among the Absonifibulinae indicating an estuary-dependent life cycle. PMID- 20027462 TI - Evaluation of HA negatively charged membranes in the recovery of human adenoviruses and hepatitis A virus in different water matrices. AB - Human adenoviruses (HAdV) and hepatitis A virus (HAV) are shed in the faeces and consequently may be present in environmental waters, resulting in an increase in pathogen concentration that can affect water quality and human health. The aim of this study was to evaluate an adsorption-elution method which utilizes negatively charged membrane HA to determine the efficient recovery of HAdV and HAV from different water matrices and to combine this procedure with a qualitative molecular method (nested RT-PCR and nested PCR). The best efficiency recovery was achieved in distilled water and treated wastewater effluent (100%) for both viruses and in recreational lagoon water for HAV (100%). The efficiency recovery was 10% for HAdV and HAV in seawater and 10% for HAdV in lagoon water. The viral detection limit by nested PCR for HAV in water samples ranged between 20-0.2 FFU/mL and 250 and 25 TCID50/mL for HAdV. In conclusion, these results suggest that the HA negatively charged membranes vary their efficiency for recovery of viral concentration depending upon the types of both enteric viruses and water matrices. PMID- 20027463 TI - Fine structure of Henneguya hemiodopsis sp. n. (Myxozoa), a parasite of the gills of the Brazilian teleostean fish Hemiodopsis microlepes (Hemiodontidae). AB - A fish-infecting myxosporean, Henneguya hemiodopsis sp. n., found infecting the gills of Hemiodopsis microlepis and collected from the Poty River near the city of Teresina, Brazil, was described based on ultrastructural studies. The parasite occurred within large whitish polysporic plasmodia (up to 200 microm in diameter) containing asynchronous developmental sporogonic stages, mainly mature spores. The spores measured 19.7 +/- 0.9 microm in total length (n = 30) and the ellipsoidal spore body was 10.8 +/- 0.5 microm long, 3.3 +/- 0.4 microm wide and 2.5 +/- 0.5 microm thick. The spores were composed of two equal shell valves adhering together along the straight suture line, with each valve having equal sized caudal tapering tails measuring 8.7 +/- 0.6 microm in length. The spores were surrounded by a thin anastomosed network of microfibrils, more evident on the tails. There were two symmetric elongated bottle-like polar capsules 3.5 +/- 0.3 microm long and 1.0 +/- 0.2 microm wide, each with a polar filament with five to six coils. Given the morphological and ultrastructural differences from previously described parasites and the specificity of the host species, we propose a new species, named H. hemiodopsis sp. n. PMID- 20027464 TI - In vitro activity of thienyl-2-nitropropene compounds against Trypanosoma cruzi. AB - The in vitro activity of four 2-nitropropene derivatives, 1-(3-benzothienyl)-2 nitropropene (N1), 1-(3-thienyl)-2-nitropropene (N2), 1-(5-bromo-2-thienyl)-2 nitropropene (N3) and 1-(4-bromo-2-thienyl)-2-nitropropene (N4), were tested against cultures of the parasite Trypanosoma cruzi. Cytotoxicity studies were performed using Vero cells. The blood trypomastigotes, amastigotes and epimastigotes showed differential degrees of sensitivity towards the four tested compounds; the highest activity against the epimastigotes and blood tripomastigotes was exhibited by N1, followed by N3, N4 and finally N2. In contrast, whereas the compounds N1, N3 and N4 exerted similar magnitudes of activity against amastigotes, N2 was found to be a much less potent compound. According to our results, the compound N1 had the highest level of activity (IC50: 0.6 microM) against epimastigotes. PMID- 20027465 TI - Treatment and seroconversion in a cohort of children suffering from recent chronic Chagas infection in Yoro, Honduras. AB - Between 1999-2002, Medecins Sans Frontieres-Spain implemented a project seeking to determine the efficacy and safety of benznidazole in the treatment of recent chronic Chagas disease in a cohort of seropositive children in the Yoro Department, Honduras. A total of 24,471 children were screened for Trypanosoma cruzi IgG antibodies through conventional enzyme-linked immunosorbent assays (ELISA) on filter paper. Recombinant ELISA (0.93% seroprevalence) showed 256 initially reactive cases, including 232 confirmed positive cases. Of these, 231 individuals were treated with benznidazole (7.5 mg/kg/day) for 60 days and were followed with a strict weekly medical control and follow-up protocol. At the end of the project, 229 patients were examined by the Honduras Secretariat of Health for post-treatment serological assessments; 88.2% seroconverted after 18 months and 93.9% seroconverted after three years. No differences were found in the seroconversion rates according to age or sex. Most of the side effects of the treatment were minor. These results support the argument that in areas where T. cruzi I is predominant and in areas affected by T. cruzi II, when vector transmission has been interrupted, Chagas disease diagnosis and treatment are feasible, necessary and ethically indisputable. PMID- 20027466 TI - Genus-specific kinetoplast-DNA PCR and parasite culture for the diagnosis of localised cutaneous leishmaniasis: applications for clinical trials under field conditions in Brazil. AB - The positivities of two methods for the diagnosis of localised cutaneous leishmaniasis (CL) were estimated in 280 patients enrolled in a clinical trial. The trial was conducted in an endemic area of Leishmania (Viannia) braziliensis and trial participants were patients with skin ulcers and positive leishmanin skin tests. Patients underwent aspirative skin punctures of the ulcerated lesions and lymph nodes for in vitro cultures, which were processed under field conditions at the local health centre. Skin lesion biopsies were tested at a reference laboratory using kinetoplastid DNA (kDNA)-PCR to detect DNA. The median time required to obtain a positive culture from the skin samples was seven days and the contamination rate of the samples was 1.8%. The positivities of the cultures from skin lesions, kDNA-PCR and the combination of the two methods were 78.2% (95% CI: 73-82.6%), 89.3% (95% CI: 85.1-92.4%) and 97.1% (95% CI: 94.5 98.5%). We conclude that parasite culture is a feasible method for the detection of Leishmania in field conditions and that the combination of culture and PCR has a potential role for the diagnosis of CL in candidates for clinical trials. PMID- 20027467 TI - Molecular and antigenic characterisation of ribosomal phosphoprotein P0 from Babesia bovis. AB - Babesia bovis is a tick-borne pathogen that remains an important constraint for the development of cattle industries in tropical and subtropical regions of the world. Effective control can be achieved by vaccination with live attenuated phenotypes of the parasite. However, these phenotypes have a number of drawbacks, which justifies the search for new, more efficient immunogens based mainly on recombinant protein technology. In the present paper, ribosomal phosphoprotein P0 from a Brazilian isolate of B. bovis was produced and evaluated with regard to conservation and antigenicity. The protein sequence displayed high conservation between different Brazilian isolates of B. bovis and several Apicomplexa parasites such as Theileria, Neospora and Toxoplasma. IgG from cattle experimentally and naturally infected with B. bovisas well as IgG1 and IgG2 from naturally infected cattle reacted with the recombinant protein. IgG from cattle experimentally infected with Babesia bigemina cross-reacted with B. bovis recombinant P0. These characteristics suggest that P0 is a potential antigen for recombinant vaccine preparations against bovine babesiosis. PMID- 20027468 TI - RFLP analysis of a PCR-amplified fragment of the 16S rRNA gene as a tool to identify Enterococcus strains. AB - Restriction fragment length polymorphism (RFLP) analysis of a PCR-amplified fragment of the 16S rRNA gene was performed on reference strains belonging to 21 different enterococcal species and on 75 Enterococcus isolates recovered from poultry meat, pasteurised milk and fresh cheese. PCR amplification generated a 275 bp fragment, which was digested with three restriction endonucleases (DdeI, HaeIII, HinfI). The strains were divided into five groups (groups A-E) on the basis of their restriction patterns. Five biochemical tests (arabinose, arginine, manitol, methyl-beta-D-glucopyranoside and raffinose) were then performed in addition to RFLP analysis to narrow the identification of enterococcal strains to the species level. PCR-RFLP, in conjunction with the selected biochemical tests, allowed the precise identification of the 21 species of Enterococcus included in the present study. This proposed method is relatively simple and rapid and can be useful as an adjunct tool for accurate identification of Enterococcus. PMID- 20027469 TI - Characterisation of pvmdr1 and pvdhfr genes associated with chemoresistance in Brazilian Plasmodium vivax isolates. AB - Plasmodium vivax control is now being hampered by drug resistance. Orthologous Plasmodium falciparum genes linked to chloroquine or sulfadoxine-pyrimethamine chemoresistance have been identified in P. vivax parasites, but few studies have been performed. The goal of the present work is to characterise pvmdr1 and pvdhfr genes in parasite isolates from a Brazilian endemic area where no molecular investigation had been previously conducted. The pvmdr1 analysis revealed the existence of single (85.7%) and double (14.3%) mutant haplotypes, while the pvdhfr examination showed the presence of double (57.2%) and triple (42.8%) mutant haplotypes. The implications of these findings are discussed. PMID- 20027470 TI - Comparative descriptions of eggs from three species of Rhodnius (Hemiptera: Reduviidae: Triatominae). AB - The authors describe and compare the morphological and ultrastructural characteristics of eggs from the three most recent described species of the genus Rhodnius Stal, 1859, which have not previously been studied. These species are Rhodnius colombiensis (Mejia, Galvao & Jurberg 1999), Rhodnius milesi (Carcavallo, Rocha, Galvao & Jurberg 2001) and Rhodnius stali (Lent, Jurberg & Galvao 1993). The results revealed that there are similarities in the exochorial architecture of optical microscopy and scanning electron microscopy; these include the predominance of hexagonal cells that are common to all Rhodnius species and variable degrees of lateral flattening, which is common not only to species of this genus, but also to the Rhodniini tribe. Differences in overall colour, the presence of a collar in R. milesi, a longitudinal bevel in R. stali and the precise length of R. colombiensis can be useful distinguishing features. As a result of this study, the key for egg identification proposed by Barata in 1981 can be updated. PMID- 20027471 TI - Vector density and the control of kala-azar in Bihar, India. AB - Bihar, India has been in the grip of kala-azar for many years. Its rampant and severe spread has made life miserable in most parts of the state. Such conditions require a comprehensive understanding of this affliction. The numbers coming out of the districts prone to the disease in the north and south Ganges have provided us with several startling revelations, as there are striking uniformities on both sides, including similar vegetation, water storage facilities, house construction and little change in risk factors. The northern areas have been regularly sprayed with DDT since 1977, but eradication of the disease appears to be a distant dream. In 2007 alone, there were as many as 37,738 cases in that region. In contrast, the southern districts of Patna and Nalanda have never had the disease in its epidemic form and endemic disease has been present in only some pockets of the two districts. In those cases, two rounds of spraying with DDT had very positive results, with successful control and no new established foci. In addition, an eleven-year longitudinal study of the man hour density and house index for the vector Phlebotomus argentipes demonstrated that they were quite high in Patna and Nalanda and quite low in north Bihar. Given these facts, an attempt has been made to unravel the role of P. argentipes saliva (salivary gland) in the epidemiology of kala-azar. It was determined that patchy DDT spraying should be avoided for effective control of kala-azar. PMID- 20027472 TI - Trypanosoma cruzi: parasite antigens sequestered in heart interstitial dendritic cells are related to persisting myocarditis in benznidazole-treated mice. AB - We investigated whether sequestered Trypanosoma cruzi antigens found in heart interstitial dendritic cells (IDCs) contribute to the residual myocarditis found in mice following treatment with benznidazole, a specific chemotherapeutic drug. IDCs are antigen-presenting cells that are MHC-II-receptor dependent. Swiss mice were divided into two experimental groups: the 1st group was infected with the Colombian strain of T. cruzi, which is resistant to treatment with benznidazole, and the 2nd group was infected with clone 21SF-C 3, which has a medium susceptibility to the drug. Treatment of the Colombian strain group started on the 120th day post-infection and for the 21SF-C3 strain group treatment was started on the 90th day. In both groups, treatment lasted for 90 days. The animals were sacrificed either 150 or 200 days post-treatment. The myocardium was analysed by immunohistochemistry using anti-MAC3, 33D1, CD11b and CD11c monoclonal antibodies for IDCs or anti-T. cruzi purified antibodies. Parasite antigens were expressed on the IDC membranes in both treated and untreated mice. Myocarditis subsided following treatment, evidenced by both histological and morphometrical evaluation. A reduction in the number of IDCs carrying T. cruzi antigens in the treated group indicates that the elimination of parasites influences antigen presentation with concomitant decreases in inflammation. There is a correlation between the presence of T. cruzi antigens in these cells and the chronic focal, residual myocarditis seen in treated mice. PMID- 20027473 TI - Influence of blood meal and mating in reproduction patterns of Triatoma brasiliensis females (Hemiptera: Reduviidae) under laboratory conditions. AB - The influence of blood meal and mating on Triatoma brasiliensis (Neiva) female fecundity, fertility, life-span and the preoviposition period were investigated under laboratory conditions. Nourishment increased fecundity, fertility and adult lifespan, whereas mating increased fecundity, fertility and decreased the preoviposition period. Females also required more than one mating to reach their full reproductive potential. Results indicate that both nourishment and mating are important in T. brasiliensis proliferation. Such information will help towards developing effective control strategies of this vector of Chagas disease. PMID- 20027474 TI - Molluscicidal activity of Hammada scoparia (Pomel) Iljin leaf extracts and the principal alkaloids isolated from them against Galba truncatula. AB - The molluscicidal activity of Hammada scoparia leaf extracts and the principal alkaloids isolated from them (carnegine and N-methylisosalsoline) were tested against the mollusc gastropod, Galba truncatula, the intermediate host of Fasciola hepatica in Tunisia. The results indicated that the molluscicidal activity was correlated with the presence of alkaloids. A significant molluscicidal value, according to the World Health Organization, was found with the methanol extract (LC50 = 28.93 ppm). Further fractionation of the methanolic extract led to the isolation of two principal alkaloids: carnegine and N methylisosalsoline. These alkaloids are isoquinolines that have not previously been characterised for their molluscicidal activity. The N-methylisosalsoline possesses the highest molluscicidal activity (LC50 = 0.47 microM against G. truncatula). PMID- 20027475 TI - Anti-mycobacterial treatment reduces high plasma levels of CXC-chemokines detected in active tuberculosis by cytometric bead array. AB - Chemokines recruit and activate leukocytes, assisting granuloma formation. Herein, we evaluated plasma chemokines in patients with active tuberculosis (ATB) and after completing treatment (TTB) and compared them to BCG-vaccinated healthy controls (HC). Levels of chemokines were measured by cytometric bead array. Levels of CXCL8, CXCL9 and CXCL10 were higher in ATB patients compared to HC, but they decreased in TTB. Levels of CCL2 and CCL5 in ATB patients were similar to those observed in HC. Thus, the high levels of CXC-chemokines detected during ATB, which can modulate the trafficking of immune cells from the periphery to the site of infection, were reversed by anti-mycobacterial treatment. PMID- 20027476 TI - Tracing lineage by phenotypic and genotypic markers in Salmonella enterica subsp. enterica serovar 1,4,[5],12:i:- and Salmonella Typhimurium isolated in state of Sao Paulo, Brazil. AB - Fifty-three Salmonella 1,4,[5],12:i:- and 45 Salmonella Typhimurium strains were characterised using phage typing, plasmid profiles and pulsed-field gel electrophoresis (PFGE) for comparison. The majority of the strains were subdivided into definitive type (DT) 41 (22.6%) and DT 193 (18%) and the 60-MDa plasmid was detected in 94.3% and 84.4% of strains, respectively. Genetic diversity was observed among all strains and 90% presented a > 70% similarity through PFGE analysis. These results suggest a close relationship between Salmonella 1,4,[5],12:i:- and Salmonella Typhimurium at the serotype level. PMID- 20027477 TI - Microsatellite markers for population genetic studies of the blowfly Chrysomya putoria (Diptera: Calliphoridae). AB - The investigation of the genetic variation and population structure of Chrysomya species is of great interest for both basic and applied research. However, very limited genetic information is available for this genus across its geographical distribution. Here, we describe 12 polymorphic microsatellite loci isolated from Chrysomya putoria with expected heterozygosities ranging from 0.1402-0.8312. These markers are of potential applied interest for forensic entomologists and for the characterisation of the genetic structure of C. putoria from recently colonised regions, with great promise for understanding the colonisation dynamics and spread of the genus Chrysomya in the New World. PMID- 20027478 TI - A new consensus for Trypanosoma cruzi intraspecific nomenclature: second revision meeting recommends TcI to TcVI. AB - In an effort to unify the nomenclature of Trypanosoma cruzi, the causative agent of Chagas disease, an updated system was agreed upon at the Second Satellite Meeting. A consensus was reached that T. cruzi strains should be referred to by six discrete typing units (T. cruzi I-VI). The goal of a unified nomenclature is to improve communication within the scientific community involved in T. cruzi research. The justification and implications will be presented in a subsequent detailed report. PMID- 20027479 TI - Characterization of the interdependency between residues that bind the substrate in a beta-glycosidase. AB - The manner by which effects of simultaneous mutations combine to change enzymatic activity is not easily predictable because these effects are not always additive in a linear manner. Hence, the characterization of the effects of simultaneous mutations of amino acid residues that bind the substrate can make a significant contribution to the understanding of the substrate specificity of enzymes. In the beta-glycosidase from Spodoptera frugiperda (Sfbetagly), both residues Q39 and E451 interact with the substrate and this is essential for defining substrate specificity. Double mutants of Sfbetagly (A451E39, S451E39 and S451N39) were prepared by site-directed mutagenesis, expressed in bacteria and purified using affinity chromatography. These enzymes were characterized using p-nitrophenyl beta-galactoside and p-nitrophenyl beta-fucoside as substrates. The k cat/Km ratio for single and double mutants of Sfbetagly containing site-directed mutations at positions Q39 and E451 was used to demonstrate that the effect on the free energy of ESdouble dagger (enzyme-transition state complex) of the double mutations (Gdouble daggerxy) is not the sum of the effects resulting from the single mutations (Gdouble daggerx and Gdouble daggery). This difference in Gdouble dagger indicates that the effects of the single mutations partially overlap. Hence, this common effect counts only once in Gdouble daggerxy. Crystallographic data on beta-glycosidases reveal the presence of a bidentate hydrogen bond involving residues Q39 and E451 and the same hydroxyl group of the substrate. Therefore, both thermodynamic and crystallographic data suggest that residues Q39 and E451 exert a mutual influence on their respective interactions with the substrate. PMID- 20027480 TI - A bovine herpesvirus 5 recombinant defective in the thymidine kinase (TK) gene and a double mutant lacking TK and the glycoprotein E gene are fully attenuated for rabbits. AB - Bovine herpesvirus 5 (BoHV-5), the agent of herpetic meningoencephalitis in cattle, is an important pathogen of cattle in South America and several efforts have been made to produce safer and more effective vaccines. In the present study, we investigated in rabbits the virulence of three recombinant viruses constructed from a neurovirulent Brazilian BoHV-5 strain (SV507/99). The recombinants are defective in glycoprotein E (BoHV-5gEDelta), thymidine kinase (BoHV-5TKDelta) and both proteins (BoHV-5gEDeltaTKDelta). Rabbits inoculated with the parental virus (N = 8) developed neurological disease and died or were euthanized in extremis between days 7 and 13 post-infection (pi). Infectivity was detected in several areas of their brains. Three of 8 rabbits inoculated with the recombinant BoHV-5gEDelta developed neurological signs between days 10 and 15 pi and were also euthanized. A more restricted virus distribution was detected in the brain of these animals. Rabbits inoculated with the recombinants BoHV 5TKDelta (N = 8) or BoHV-5gEDeltaTKDelta (N = 8) remained healthy throughout the experiment in spite of variable levels of virus replication in the nose. Dexamethasone (Dx) administration to rabbits inoculated with the three recombinants at day 42 pi did not result in viral reactivation, as demonstrated by absence of virus shedding and/or increase in virus neutralizing titers. Nevertheless, viral DNA was detected in the trigeminal ganglia or olfactory bulbs of all animals at day 28 post-Dx, demonstrating they were latently infected. These results show that recombinants BoHV-5TKDelta and BoHV-5gEDeltaTKDelta are attenuated for rabbits and constitute potential vaccine candidates upon the confirmation of this phenotype in cattle. PMID- 20027481 TI - Diphtheria toxin IgG levels in military and civilian blood donors in Rio de Janeiro, Brazil. AB - Serologic data on diseases that are preventable by vaccines are necessary to evaluate the success of immunization programs and to identify susceptible subgroups. In the present study, we determined serum IgG levels against diphtheria toxin of military and civilian blood donors (N = 75; 69.3% males and 30.7% females) aged 18-64 years, from the Brazilian Army Biology Institute, Rio de Janeiro, using a commercial diphtheria kit (Diphtheria IgG ELISA; IBL, Germany). Most (63%) unprotected military donors were from the older age group of 41 to 64 years. In contrast, the majority (71%) of young military donors (18 to 30 years) were fully protected. About half of the military donors aged 31 to 40 years were protected against diphtheria. Among the civilians, about 50% of persons aged 18 to 30 years and 31 to 40 years had protective antibody levels against diphtheria as also did 64% of individuals aged 41 to 64 years. All civilians had a similar antibody response (geometric mean = 0.55 IU/mL) independent of age group. Military donors aged 18-30 years had higher IgG levels (geometric mean = 0.82 IU/mL) than military donors of 41-64 years (geometric mean = 0.51 IU/mL; P > 0.05). In conclusion, the existence of a considerable proportion of susceptible adults supports the position that reliable data on the immune status of the population should be maintained routinely and emphasizes the importance of adequate immunization during adulthood. PMID- 20027482 TI - HFE gene mutations and iron status of Brazilian blood donors. AB - Mutations of the HFE and TFR2 genes have been associated with iron overload. HFE and TFR2 mutations were assessed in blood donors, and the relationship with iron status was evaluated. Subjects (N = 542) were recruited at the Hemocentro da Santa Casa de Sao Paulo, Sao Paulo, Brazil. Iron status was not influenced by HFE mutations in women and was independent of blood donation frequency. In contrast, men carrying the HFE 282CY genotype had lower total iron-binding capacity (TIBC) than HFE 282CC genotype carriers. Men who donated blood for the first time and were carriers of the HFE 282CY genotype had higher transferrin saturation values and lower TIBC concentrations than those with the homozygous wild genotype for the HFE C282Y mutation. Moreover, in this group of blood donors, carriers of HFE 63DD plus 63HD genotypes had higher serum ferritin values than those with the homozygous wild genotype for HFE H63D mutation. Multiple linear regression analysis showed that HFE 282CY leads to a 17.21% increase (P = 0.018) and a 83.65% decrease (P = 0.007) in transferrin saturation and TIBC, respectively. In addition, serum ferritin is influenced by age (3.91%, P = 0.001) and the HFE 63HD plus DD genotype (55.84%, P = 0.021). In conclusion, the HFE 282Y and 65C alleles were rare, while the HFE 63D allele was frequent in Brazilian blood donors. The HFE C282Y and H63D mutations were associated with alterations in iron status in blood donors in a gender-dependent manner. PMID- 20027483 TI - Rat Dlx5 is expressed in the subventricular zone and promotes neuronal differentiation. AB - The molecular mechanisms and potential clinical applications of neural precursor cells have recently been the subject of intensive study. Dlx5, a homeobox transcription factor related to the distal-less gene in Drosophila, was shown to play an important role during forebrain development. The subventricular zone (SVZ) in the adult brain harbors the largest abundance of neural precursors. The anterior SVZ (SVZa) contains the most representative neural precursors in the SVZ. Further research is necessary to elucidate how Dlx5-related genes regulate the differentiation of SVZa neural precursors. Here, we employed immunohistochemistry and molecular biology techniques to study the expression of Dlx5 and related homeobox genes Er81 and Islet1 in neonatal rat brain and in in vitro cultured SVZa neural precursors. Our results show that Dlx5 and Er81 are also highly expressed in the SVZa, rostral migratory stream, and olfactory bulb. Islet1 is only expressed in the striatum. In cultured SVZa neural precursors, Dlx5 mRNA expression gradually decreased with subsequent cell passages and was completely lost by passage four. We also transfected a Dlx5 recombinant plasmid and found that Dlx5 overexpression promoted neuronal differentiation of in vitro cultured SVZa neural precursors. Taken together, our data suggest that Dlx5 plays an important role during neuronal differentiation. PMID- 20027484 TI - Differential binding with ERalpha and ERbeta of the phytoestrogen-rich plant Pueraria mirifica. AB - Variations in the estrogenic activity of the phytoestrogen-rich plant, Pueraria mirifica, were determined with yeast estrogen screen (YES) consisting of human estrogen receptors (hER) hERalpha and hERbeta and human transcriptional intermediary factor 2 (hTIF2) or human steroid receptor coactivator 1 (hSRC1), respectively, together with the beta-galactosidase expression cassette. Relative estrogenic potency was expressed by determining the beta-galactosidase activity (EC(50)) of the tuber extracts in relation to 17beta-estradiol. Twenty-four and 22 of the plant tuber ethanolic extracts interacted with hERalpha and hERbeta, respectively, with a higher relative estrogenic potency with hERbeta than with hERalpha. Antiestrogenic activity of the plant extracts was also determined by incubation of plant extracts with 17beta-estradiol prior to YES assay. The plant extracts tested exhibited antiestrogenic activity. Both the estrogenic and the antiestrogenic activity of the tuber extracts were metabolically activated with the rat liver S9-fraction prior to the assay indicating the positive influence of liver enzymes. Correlation analysis between estrogenic potency and the five major isoflavonoid contents within the previously HPLC-analyzed tuberous samples namely puerarin, daidzin, genistin, daidzein, and genistein revealed a negative result. PMID- 20027485 TI - Immune cells and oxidative stress in the endotoxin tolerance mouse model. AB - Sepsis is a systemic inflammatory response that can lead to tissue damage and death. In order to increase our understanding of sepsis, experimental models are needed that produce relevant immune and inflammatory responses during a septic event. We describe a lipopolysaccharide tolerance mouse model to characterize the cellular and molecular alterations of immune cells during sepsis. The model presents a typical lipopolysaccharide tolerance pattern in which tolerance is related to decreased production and secretion of cytokines after a subsequent exposure to a lethal dose of lipopolysaccharide. The initial lipopolysaccharide exposure also altered the expression patterns of cytokines and was followed by an 8- and a 1.5-fold increase in the T helper 1 and 2 cell subpopulations. Behavioral data indicate a decrease in spontaneous activity and an increase in body temperature following exposure to lipopolysaccharide. In contrast, tolerant animals maintained production of reactive oxygen species and nitric oxide when terminally challenged by cecal ligation and puncture (CLP). Survival study after CLP showed protection in tolerant compared to naive animals. Spleen mass increased in tolerant animals followed by increases of B lymphocytes and subpopulation Th1 cells. An increase in the number of stem cells was found in spleen and bone marrow. We also showed that administration of spleen or bone marrow cells from tolerant to naive animals transfers the acquired resistance status. In conclusion, lipopolysaccharide tolerance is a natural reprogramming of the immune system that increases the number of immune cells, particularly T helper 1 cells, and does not reduce oxidative stress. PMID- 20027486 TI - Role of 11beta-hydroxysteroid dehydrogenase 2 renal activity in potassium homeostasis in rats with chronic renal failure. AB - Aldosterone concentrations vary in advanced chronic renal failure (CRF). The isozyme 11beta-hydroxysteroid dehydrogenase 2 (11beta-HSD2), which confers aldosterone specificity for mineralocorticoid receptors in distal tubules and collecting ducts, has been reported to be decreased or normal in patients with renal diseases. Our objective was to determine the role of aldosterone and 11beta HSD2 renal microsome activity, normalized for glomerular filtration rate (GFR), in maintaining K+ homeostasis in 5/6 nephrectomized rats. Male Wistar rats weighing 180-220 g at the beginning of the study were used. Rats with experimental CRF obtained by 5/6 nephrectomy (N = 9) and sham rats (N = 10) were maintained for 4 months. Systolic blood pressure and plasma creatinine (Pcr) concentration were measured at the end of the experiment. Sodium and potassium excretion and GFR were evaluated before and after spironolactone administration (10 mg.kg-1.day-1 for 7 days) and 11beta-HSD2 activity on renal microsomes was determined. Systolic blood pressure (means +/- SEM; Sham = 105 +/- 8 and CRF = 149 +/- 10 mmHg) and Pcr (Sham = 0.42 +/- 0.03 and CRF = 2.53 +/- 0.26 mg/dL) were higher (P < 0.05) while GFR (Sham = 1.46 +/- 0.26 and CRF = 0.61 +/- 0.06 mL/min) was lower (P < 0.05) in CRF, and plasma aldosterone (Pald) was the same in the two groups. Urinary sodium and potassium excretion was similar in the two groups under basal conditions but, after spironolactone treatment, only potassium excretion was decreased in CRF rats (sham = 0.95 +/- 0.090 (before) vs 0.89 +/- 0.09 microEq/min (after) and CRF = 1.05 +/- 0.05 (before) vs 0.37 +/- 0.07 microEq/min (after); P < 0.05). 11beta-HSD2 activity on renal microsomes was lower in CRF rats (sham = 0.807 +/- 0.09 and CRF = 0.217 +/- 0.07 nmol.min-1.mg protein-1; P < 0.05), although when normalized for mL GFR it was similar in both groups. We conclude that K+ homeostasis is maintained during CRF development despite normal Pald levels. This adaptation may be mediated by renal 11beta-HSD2 activity, which, when normalized for GFR, became similar to that of control rats, suggesting that mineralocorticoid receptors maintain their aldosterone selectivity. PMID- 20027487 TI - Lung hyperinflation stimulates the release of inflammatory mediators in spontaneously breathing subjects. AB - Lung hyperinflation up to vital capacity is used to re-expand collapsed lung areas and to improve gas exchange during general anesthesia. However, it may induce inflammation in normal lungs. The objective of this study was to evaluate the effects of a lung hyperinflation maneuver (LHM) on plasma cytokine release in 10 healthy subjects (age: 26.1 +/- 1.2 years, BMI: 23.8 +/- 3.6 kg/m(2)). LHM was performed applying continuous positive airway pressure (CPAP) with a face mask, increased by 3-cmH(2)O steps up to 20 cmH(2)O every 5 breaths. At CPAP 20 cmH(2)O, an inspiratory pressure of 20 cmH(2)O above CPAP was applied, reaching an airway pressure of 40 cmH(2)O for 10 breaths. CPAP was then decreased stepwise. Blood samples were collected before and 2 and 12 h after LHM. TNF alpha, IL-1beta, IL-6, IL-8, IL-10, and IL-12 were measured by flow cytometry. Lung hyperinflation significantly increased (P < 0.05) all measured cytokines (TNF-alpha: 1.2 +/- 3.8 vs 6.4 +/- 8.6 pg/mL; IL-1beta: 4.9 +/- 15.6 vs 22.4 +/- 28.4 pg/mL; IL-6: 1.4 +/- 3.3 vs 6.5 +/- 5.6 pg/mL; IL-8: 13.2 +/- 8.8 vs 33.4 +/ 26.4 pg/mL; IL-10: 3.3 +/- 3.3 vs 7.7 +/- 6.5 pg/mL, and IL-12: 3.1 +/- 7.9 vs 9 +/- 11.4 pg/mL), which returned to basal levels 12 h later. A significant correlation was found between changes in pro- (IL-6) and anti-inflammatory (IL 10) cytokines (r = 0.89, P = 0.004). LHM-induced lung stretching was associated with an early inflammatory response in healthy spontaneously breathing subjects. PMID- 20027488 TI - Effects of fluoxetine and imipramine in rat fetuses treated during a critical gestational period: a macro and microscopic study. AB - OBJECTIVE: To evaluate morphological alterations in rat fetuses treated with fluoxetine and imipramine during the "critical" period of gestation. METHOD: Fifteen female rats were separated into three groups (n = 5) and treated with 10 mg/kg/day of test substances on the ninth, tenth and eleventh day of pregnancy: G1, fluoxetine; G2, imipramine hydrochloride; G3 (control), saline. On day 21, cesarean sections were performed to release the fetuses, whose bodies were weighed and macroscopically analyzed. The placenta was also weighed. The fetuses were then fixed and their encephala removed and weighed. Sections of the frontal lobe were taken for histological neuron counting. RESULTS: G1 and G2 showed the highest fetal body weight. Placental weight showed statistical differences (p < 0.01): G1 weighed more than G2 and G3. Otherwise, G2 exhibited the highest encephalon weight, statistically differing from G3 (control) and fluoxetine treated G1 (p < 0.01). However, G1 did not statistically (p > 0.01) differ from the control group. G3 showed the highest number of neurons per area when compared to G1 and G2 (p < 0.01). CONCLUSION: The use of antidepressants in rats caused an increase in fetal weight and a decrease in the number of fetal frontal lobe neurons, thus suggesting that the use of antidepressants by pregnant women can induce depression in fetuses due to alterations in their neural development. PMID- 20027489 TI - Short admission in an emergency psychiatry unit can prevent prolonged lengths of stay in a psychiatric institution. AB - OBJECTIVE: Characterize and compare acute psychiatric admissions to the psychiatric wards of a general hospital (22 beds), a psychiatric hospital (80) and of an emergency psychiatry unit (6). METHOD: Survey of the ratios and shares of the demographic, diagnostic and hospitalization variables involved in all acute admissions registered in a catchment area in Brazil between 1998 and 2004. RESULTS: From the 11,208 admissions, 47.8% of the patients were admitted to a psychiatric hospital and 14.1% to a general hospital. The emergency psychiatry unit accounted for 38.1% of all admissions during the period, with a higher variability in occupancy rate and bed turnover during the years. Around 80% of the hospital stays lasted less than 20 days and in almost half of these cases, patients were discharged in 2 days. Although the total number of admissions remained stable during the years, in 2004, a 30% increase was seen compared to 2003. In 2004, bed turnover and occupancy rate at the emergency psychiatry unit increased. CONCLUSION: The increase in the number of psychiatric admissions in 2004 could be attributed to a lack of new community-based services available in the area beginning in 1998. Changes in the health care network did affect the emergency psychiatric service and the limitations of the community-based network could influence the rate of psychiatric admissions. PMID- 20027490 TI - [Effects of exposure to altitude on neuropsychology aspects: a literature review]. AB - OBJECTIVE: Discuss the effects of altitude exposure on neuropsychological functions. METHOD: We have conducted a literature review using as source indexed articles at Pubmed in the period from 1921 to 2008, using the following key words: 'cognition and hypoxia', 'hypoxia and neuropsychology', 'acute hypoxia', 'chronic hypoxia', and 'acclimatization and hypoxia', as well as specific books on the subject. DISCUSSION: Acute and chronic effects of Hypoxia can alter many of the neuropsychological functions in different altitudes due to physiological changes resulted by the oxygen (O2) partial decrease that can lead to neuropsychological alterations in individuals exposed to high altitudes. CONCLUSION: Individuals exposed to high altitudes must use an O2 supplementation and the practice of acclimatization, among other strategy ways that can be used in order to minimize the negative effects of hypoxia on neuropsychological aspects. PMID- 20027491 TI - Women's perceptions and practices regarding prevention and health promotion in primary healthcare. AB - OBJECTIVE: To analyze the perceptions and participation of female basic health unit users with regard to prevention and health promotion. METHODOLOGICAL PROCEDURES: Qualitative study with 20 users of a family health unit in the municipality of Belo Horizonte, Southeastern Brazil, in 2007. The interview script included questions about the health-disease process and prevention and health promotion. The content analysis technique was used to analyze the reports. RESULTS ANALYSIS: The perception of disease prevention was influenced by Leavell & Clark's theory, which is expressed as actions that avoid the appearance, progression or worsening of disease. Health promotion was regarded as a level of prevention and associated with the individual responsibility and the positive concept of health. Prevention and health promotion practices were influenced by the positive health concept, the possibility of causing pleasure/displeasure, the way in which they might interfere in daily life, by the concept of will-power and the value attributed to life. CONCLUSIONS: The discourse about disease prevention and health promotion is marked by traditional concepts. However, the inclusion of the positive health concept, allied to pleasure and will-power, are the main behavior determinants. Strategies based on a more comprehensive approach to the health-disease process are needed, thus reflecting the modern principles of health promotion. PMID- 20027492 TI - [Scientific evidence: information management]. PMID- 20027493 TI - Public health and the knowledge industry. AB - Knowledge plays an important role in health care. The production and diffusion of health-related knowledge are increasingly under the control of private commercial interests, which are characterized by conflicts of interests that result in abuses of power. Considerable research has been done on the medical-industrial complex and its role in the production of power imbalances and the consequent abuses, but little attention has been dedicated to the role played by the publishing industry, which can be subject to the same problems. The widely diffused idea that 'frequent and major changes' occur in medicine, albeit unsupported by clearcut evidence, is an effective marketing tool for both the pharmaceutical and publishing industries, who feed and thrive on physicians' insecurities. The production and distribution of knowledge should be addressed as a strategic component of public health. PMID- 20027494 TI - An outbreak of bat-transmitted human rabies in a village in the Brazilian Amazon. AB - During 45 days without electrical power, 57 individuals (8.7% of the population) from the village of Antonio Dino (municipality of Turiacu, Northeastern Brazil) were attacked by bats and 16 died from human rabies. The aim of the study was to analyze the factors associated with bat attacks and the development of human rabies. Of the 46 individuals, who suffered bat attacks, 36 (78.3%) were under 17 years of age. The risk factors associated with bat attacks were age under 17 years, having observed bats inside the bedroom and having been without electrical power in the house. Age under 17 years and having been without electrical power in the house were factors associated with human rabies. PMID- 20027495 TI - Distribution of Lutzomyia whitmani in phytoregions of the state of Maranhao, Northeastern Brazil. AB - The study had the aim of characterizing the geographical distribution of Lutzomyia whitmani s.l. in the state of Maranhao, Northeastern Brazil. Between 1992 and 2005, 9,600 specimens (65.1% males and 34.9% females) were caught in the rural and urban zones of 35 municipalities in regions consisting of forests, savanna and mixed vegetation with coconut plantations, sandbanks and heath. Greater abundance was observed in areas surrounding dwellings (91.6%) than inside the dwellings (8.4%). The presence of the vector in different phytoregions and in rural and urban areas favors the transmission of tegumentary leishmaniasis in these environments. This taxon may constitute a complex of species in Maranhao, which can be confirmed by molecular biology studies. PMID- 20027496 TI - Physical activity by pregnant women and outcomes for newborns: a systematic review. AB - A systematic review was carried out aiming at analyzing daily physical activity during pregnancy and the outcomes of birth weight, prematurity, and intrauterine growth restriction. Of 52 articles indexed in Medline, 22 that showed better methodological quality were included. Among the 22 articles analyzed, only two did not detect a significant association between physical activity and the outcomes studied. There was large variation between the indicators of maternal physical activity, which included occupational, household, recreational and, all or some, locomotive activities. Among ten articles that measured total daily physical activity, only one article did not find any association. The results support the hypothesis that both excessive and insufficient physical activity impact negatively on pregnancy outcomes. PMID- 20027497 TI - Time trend in breast-feeding in the city of Rio de Janeiro, Southeastern Brazil: 1996-2006. AB - OBJECTIVE: To analyze time trend in breast-feeding (BF) and exclusive breast feeding (EBF). METHODS: Data from a monitoring system, based on surveys conducted during the National Immunization Campaign in the city of Rio de Janeiro, Southeastern Brazil, in 1996, 1998, 2000, 2003 and 2006, were analyzed. Study population was comprised of 19,044 children younger than one year of age, who were present in vaccination stations. A probability cluster sample (vaccination stations), self-weighted and representative of the population of children younger than 12 months of age (<12), was studied for each year. A structured questionnaire with closed questions about the child's diet at the moment of the study and maternal sociodemographic characteristics was applied. The BF and EBF indicators proposed by the World Health Organization were adopted. RESULTS: BF<12 increased from 61.3% to 73.4% between 1996 and 2006. Similar trend was observed in all age groups analyzed. EBF in children <4 and <6 months of age (EBF<6) increased from 18.8% to 42.4% and from 13.8% to 33.3%, respectively. Improvements in BF>6 and EBF<6 were found in all categories of all maternal sociodemographic variables. For EBF<6, the disadvantage observed in women with a lower level of education in 1996 and in women who worked in 1998 was not completely overcome by 2006. CONCLUSIONS: BF and EBF increased in the period studied, independently from child age group and maternal socio-demographic characteristics. The differences found among women in distinct sociodemographic situations were not completely overcome. PMID- 20027498 TI - [Efficacy and safety of the Brazilian vaccine against hepatitis B in newborns]. AB - OBJECTIVE: To analyze the efficacy and safety of a recombinant Hepatitis B vaccine in newborns. METHODS: The study was carried out in a general hospital in the city of Guarulhos, Southeastern Brazil, between 2002 and 2005. The recombinant Hepatitis B vaccine from Instituto Butantan (VrHB-IB) was tested in two clinical trials. In both trials, newborns were randomly allocated to the experimental or control (reference vaccine) groups. Newborns were given three doses of vaccine, one up to 24 hours after birth and the other two 30 and 180 days later. In the first trial, 538 newborns completed the immunization protocol, and 486 in the second. Vaccines were considered equivalent when seroprotection difference was below 5%. RESULTS: Seroprotection in the first trial (anti-HBs > or = 10mUI/ml) was 92.5% (247/267) in the experimental group, compared to 98.5% (267/271) in the control (p = 0.001). With this result, VrHB-IB did not fulfill the pre-established criterion for equivalence. After increasing the concentration of antigen in the vaccine to 25 microg, seroprotection reached 100% in the experimental group and 99.2% in the control. No severe adverse effects were recorded. CONCLUSIONS: The reformulated VrHB-IB is considered equivalent to the reference vaccine, and its use is recommended in newborns. PMID- 20027499 TI - Characteristics of healthcare services associated with adherence to tuberculosis treatment. AB - OBJECTIVE: To analyze characteristics related to adherence to tuberculosis treatment in tuberculosis outpatient clinics. METHODS: An ecological study was conducted in outpatient clinics for the treatment of tuberculosis cases in the sanitary districts of Salvador, Northeastern Brazil, in 2006. The sample was composed of the municipal health units that assisted 67.2% of the 2,283 tuberculosis cases that were reported during the year. The following variables were analyzed: cure, dropout, tests, health team and benefits offered to the patients. Chi-square test or Fisher's exact test was used to verify the association between variables, and associations with p<0.05 were considered to be statistically significant. RESULTS: Of the studied cases, 78.4% resulted in cure, 8.6% in dropout, 2.2% in death and 8.1% in transference. Adherence rates per health unit varied between 66.7% and 98.1%. The variables cure and dropout showed a statistically significant association with adherence in the comparison of proportions. All the units with high adherence rates had complete health teams. CONCLUSIONS: Adherence was an important factor for the outcomes cure and dropout, but the index of units that achieved the cure goals was low. The presence of a complete multidisciplinary team in the tuberculosis program may help the patient understand his/her illness and contribute to adherence to treatment. PMID- 20027500 TI - Trans fatty acid intake among the population of the city of Sao Paulo, Brazil. AB - OBJECTIVE: To analyze the monounsaturated and polyunsaturated trans fatty acid intake among the general population. METHODS: A cross-sectional study was conducted in Sao Paulo, Southeastern Brazil, in 2003, on a representative sample of 2,298 male and female subjects, including 803 adolescents (12 to 19 years), 713 adults (20 to 59 years) and 782 elderly people (60 years or over). Food intake was measured using 24-hour recall. Mean trans fatty acid intake was described according to gender and age group. RESULTS: The mean trans fatty acid intake was 5.0 g/day (SE = 0.1), accounting for 2.4% (SE = 0.1) of total energy and 6.8% (SE = 0.1) of total lipids. The adolescents had the highest mean intake levels (7.4 g/day; 2.9% of energy) while the adults and the elderly had similar intake (2.2% of energy for both; 6.4% of lipids and 6.5% of lipids, respectively). The mean trans fatty acid intake among adult and elderly women (approximately 2.5% of energy and 7.0% of lipids) was higher than among men in the same age group. The food item with the highest contribution towards trans fatty acids was margarine, accounting for more than 30% of total intake, followed by filled cookies among adolescents and meat among adults and the elderly. CONCLUSIONS: The trans fatty acid intake is above the level recommended by the World Health Organization. Replacement of the trans fatty acids in manufactured food items may be an effective measure for reducing trans fatty acid intake in Brazil. PMID- 20027501 TI - Evaluation of enteroparasite control activities in a Kaingang community of Southern Brazil. AB - OBJECTIVE: To analyze the parasitological state of families in an indigenous community after institution of enteroparasite control measures. METHODS: A longitudinal study was conducted between 2004 and 2006 among 447 individuals of the Kaingang ethnic group, in the municipality of Candido de Abreu, Southern Brazil. The enteroparasite control measures were: sanitation improvements in 2003, antiparasite treatments that were administered during the study period, and health education activities that were started in 2005. Parasitological health indicators were obtained through three coproparasitological surveys (in 2004, 2005 and 2006), in which 250, 147 and 126 stool samples, respectively, were collected. These were evaluated using the spontaneous sedimentation, centrifugation-flotation and Kato-Katz methods. Housing and hygiene conditions were assessed by means of a questionnaire applied to 69 (2004), 57 (2005) and 38 (2006) of the 90 families. RESULTS: The overall prevalences of enteroparasites were 91.6% (2004), 94.6% (2005) and 87.3% (2006) and did not show any significant reduction. The prevalence of some species decreased, while the prevalence of others increased significantly. High-intensity infections due to geohelminths presented rates of less than 2% over the study period. The proportion of the interviewees who reported using a toilet increased from 38.8% to 71.1% (p< 0.005) and the proportion taking antiparasite agents increased from 70.2% to 100% (p= 0.001). CONCLUSIONS: There were significant improvements in the parasitological health indicators, such as reductions in the prevalence of some species of enteroparasites and maintenance of a low parasite load, thus showing the importance of combining antiparasite treatment with sanitation improvements. PMID- 20027502 TI - Perception of the environment and leisure-time physical activity in the elderly. AB - OBJECTIVE: To analyze the association between leisure-time physical activity and perception of the environment in the elderly. METHODS: Cross-sectional study performed with 385 elderly individuals aged 60 years or older, living in the district of Ermelino Matarazzo, in the city of Sao Paulo, Southeastern Brazil, in 2007. The long version of the International Physical Activity Questionnaire, in addition to specific questions for the study, was used to assess leisure-time physical activity. Assessment of the environment was performed using a perception scale adapted from the Neighborhood Environment Walkability Scale. For statistical analysis, multiple logistic regression models were stratified by sex and controlled by level of education. The cut-off point of 150 minutes of physical activity per week was used to classify individuals who were active during leisure time. RESULTS: The proportion of elderly individuals active during leisure time was 15.2% (19.1% and 12.5% for men and women, respectively). Presence of sports courts (OR=2.95), banks (OR=3.82) and health clinics (OR=3.60), good perception of safety during the day (OR=4.21) and invitation from friends to exercise (OR=3.13) were associated with leisure-time physical activity in men. Presence of churches or religious temples (OR=5.73), gyms (OR=2.49) and squares (OR=3.63) were associated with leisure-time physical activity in women. CONCLUSIONS: Programs to promote physical activities for the elderly population must consider the variables associated with public and private structures (gyms, squares, sports courts, health clinics and banks), places where there are social gatherings (churches), social support (invitation from friends to exercise) and perception of safety. PMID- 20027503 TI - Hospitalizations sensitive to primary care as an evaluation indicator for the Family Health Strategy. AB - OBJECTIVE: To identify variables associated with hospitalizations sensitive to primary care. METHODS: A hospital morbidity survey was conducted using a random sample of 660 patients hospitalized in clinical and surgical wards of hospitals that had service agreements with the Brazilian National Health System (SUS), in the municipality of Montes Claros, Southeastern Brazil, between 2007 and 2008. Interviews were held with patients and members of their families using a specific form, and the patients' medical files were investigated. The definition of conditions considered sensitive to primary care was based on the Ministry of Health's list. Associations shown by socioeconomic and health variables in relation to hospitalizations sensitive to primary care were analyzed using bivariate and multiple logistic regression analyses. RESULTS: The percentage of hospitalizations sensitive to primary care in the study group was 38.8% (n = 256). The variables that remained statistically associated with conditions considered sensitive to primary care were: previous hospitalization (OR = 1.62; 95% CI: 1.51;2.28); regular visits to healthcare units (OR = 2.20; 95% CI: 1.44;3.36); low schooling level (OR = 1.50; 95% CI: 1.02;2.20); health checks not performed by the family health team (OR = 2.48; 95% CI: 1.64;3.74); hospitalization requested by physicians who were not part of the family health team (OR = 2.25; 95% CI: 1.03;4.94); and age greater than or equal to 60 years (OR = 2.12; 95% CI: 1.45;3.09). CONCLUSIONS: The variables associated with hospitalizations sensitive to primary care are particularly those relating to patients, such as age, schooling level and previous hospitalization, but regular health checks outside of the Family Health Strategy doubled the likelihood of hospitalization. PMID- 20027504 TI - In-hospital mortality and complications after coronary angioplasty, City of Rio de Janeiro, Southeastern Brazil. AB - OBJECTIVE: To estimate in-hospital mortality and prevalence of complications of percutaneous transluminal coronary angioplasty (PTCA) in public hospitals. METHODS: Data for 2,913 PTCA were obtained from the Brazilian National Health System (SUS) Hospital Authorization Database in the city of Rio de Janeiro, Southeastern Brazil, between 1999 and 2003. After simple random sampling and data weighting, 529 medical records of patients undergoing PTCA, including all deaths, in four public hospitals (federal and state university, and federal and state reference hospitals) were studied. Comparison tests of mortality according to patient characteristics, comorbidities, complications, types of PTCA procedures, and indications for PTCA were performed using Poisson's regression models. RESULTS: The overall in-hospital mortality was 1.6% (range: 0.9-6.8%). The age distribution of mortality was as follows: 0.2% in patients younger than 50; 1.6% in those 50-69; and 2.7% in those older than 69. High mortality was seen in primary and rescue PTCAs: 17.4% and 13.1%, respectively; and mortality in elective PTCA was 0.8%. The main complications during PTCA were dissection (5%; mortality: 11.5%) and artery occlusion (2.6%; mortality: 21.8%). Bleeding was seen in 5.9% of the patients (mortality: 5.6%) and 3.0% required blood transfusion (mortality: 12.0%). The complication of acute myocardial infarction was seen in 1.1% of patients (mortality: 38%) and stroke was associated with a mortality of 17.5%. CONCLUSIONS: The cardiac in-hospital mortality was high when PTCA was performed for a patient with ST elevation acute myocardial infarction. Elective PTCA had mortality and complications levels above the expected in four public hospitals in the main city of Rio de Janeiro. PMID- 20027505 TI - An epidemic in times of globalisation. PMID- 20027506 TI - [Factors related to using health services in Costa Rica]. AB - OBJECTIVE: Determining Costa Ricans' behaviour patterns when using health services. METHODS: The Costa Rican Health Survey was used. Central tendency, dispersion, percentages, frequencies, Ji-Square and Kruskal-Walis test measurements were analysed. RESULTS: Area of residence and income level were the predisposing factors in both out-patient and emergency services whereas educational level was so in terms of hospitalisation service. Health insurance status and the area of residence were the enabling factors associated with using out-patient and hospitalisation services. The need factors associated with outpatient services were the individuals' perceived state of health, having remained in bed at least until noon and suffering some chronic disease; chronic disease was associated with hospitalisation and remaining in bed when using the emergency service. DISCUSSION: Most variables postulated by Andersen and Newman's model as being determinants for using health services were present in Costa Rica. Nevertheless, the significance of its factors varied between outpatient, hospitalisation and emergency services. PMID- 20027507 TI - [Professional opinion about hospitalising Latin-American immigrant children in Andalucia, Spain]. AB - OBJECTIVE: There has been an increase in paediatric hospital attention being provided for Latin-American immigrant patients in Spain during the recent years. This work was aimed at ascertaining professionals' opinion regarding this population's specific and differential needs concerning Latin-American immigrant children being hospitalised. METHODS: The study consisted of a qualitative, phenomenological type of investigation, based on semi-structured interviews of key informers in Andalusia (Spain). The subjects of the study were professionals from paediatric services in hospital centres in Andalusia. RESULTS: One of the most prominent (positive) results revealed that the Latin-American immigrant population presented less difficulties regarding hospital functioning when compared to other immigrant groups. CONCLUSIONS: The psychological aspects which had most impact on paediatric hospitalisation would be emotional alterations, such as the presence of anxiety and stress, or other aspects, like a lack of trust when being hospitalised. PMID- 20027508 TI - [Perceptions of pregnant and postpartum women's feelings about preeclampsia]. AB - OBJECTIVE: The present study was aimed at understanding how pregnant and postpartum women with preeclampsia perceived and experienced their condition. MATERIALS AND METHODS: Interviews and observation were conducted with 20 women having preeclampsia between February and June 2007 in the outpatients department and hospitalisation ward of a public maternity hospital in north-eastern Brazil. The analysis was based on Gadamer's hermeneutics, with construction of thematic categories. RESULTS: The women were aware of the gravity and some of the risks to which they were exposed; however, they had little knowledge about preeclampsia and its consequences and little information was given by the health professionals who were treating them. The most present feeling among these women was fear of death, fear of miscarriage and fear caused by lack of information about the disease. CONCLUSIONS: The perception of the gravity of the disease coupled to the little information given during treatment increased fear amongst these women. Health professionals making high risk prenatal assistance more humanised (especially regarding preeclampsia) could facilitate an approach which would take the disease's emotional dimension into account during pregnant and postpartum women's clinical appointments. PMID- 20027509 TI - [Effects of differerent types of training and life styles on anthropometric and cardiocirculatory markers in aging]. AB - OBJECTIVE: The aim of this experimental study was to compare the effects of different forms of training and life styles on hemodynamic markers, cardiac capacity, Waist-to-Hip Ratio-WHR and Body Mass Index-BMI in elders, between 60-69 years, of both sexes. MATERIALS AND METHODS: The sample was composed of elders, between 60-69 years, of both sexes: masculine N=159 and feminine N=157. They were divided into three groups: experimental 1: n=53 (masculine) and n=51 (feminine) activity: cardio respiratory endurance; experimental 2: n=53 (masculine) and n=53 (feminine) activity: resistance training with Local Muscle Resistance-RML; control group: n=53 (masculine) and n=53 (feminine) who didn t practice any regular physical activity, considered sedentary. The variables investigated were BMI, WHR, Cardiac Frequency at rest, Systolic Blood Pressure, Diastolic Blood Pressure, Mean Blood Pressure and Double Product. The data analysis was descriptive and inferential and comparisons were performed between groups and inter groups. The data collected in this study and the hypothesis test showed the benefits of physical activity on the selected parameters for observation. RESULTS: It was demonstrated the existence of significative difference (p<0.05) in the studied variables. CONCLUSIONS: There is a need for adherence in broad spectrum physical activity in elders. PMID- 20027510 TI - Tuberculosis control and the private sector in a low incidence setting in Argentina. AB - OBJECTIVES: Determining private doctors' (PD) knowledge and practice regarding tuberculosis (TB) control issues and exploring the availability of TB drugs in private pharmacies (PP) in a low incidence setting in Argentina (Santa Fe). METHODS: A questionnaire was applied to random sample of 87 PPs and 61 PDs, proportionally assigned according to the local distribution of specialists and general practitioners. Their knowledge and practice were explored regarding the symptoms, diagnosis and case management of pulmonary TB, as defined and specified by Argentinean TB management guidelines. RESULTS: Although most PDs were aware of the main symptoms presented by TB, they were unlikely to ask for a microscope examination for a person suffering persistent cough. More than 30 % of PDs considered X-ray before 'microscope examination of stained sputa' for TB diagnosis. PDs had significantly lower levels of knowledge regarding the symptoms and diagnostic procedures for TB compared to those who combined public and private work. Most PDs (91,3 %) referred their TB patients to a public facility for treatment. Accordingly, very few TB drug prescriptions were managed in PPs. DISCUSSION: Addressing mismanagement of TB patients in the private sector does not appear to be a problem in Santa Fe in terms of treatment (although being considered crucial for effective TB control) because most PDs are aware of the TB programme which appears to be comprehensive and accessible. The private sector's main aim should be to improve its skills regarding TB diagnosis. PMID- 20027511 TI - [Cardiometabolic factors in a comunity located at Valencia city, Venezuela]. AB - OBJECTIVE: Assessing the prevalence of cardiometabolic factors in residents from a community in the north of Valencia city, Venezuela, and establishing clustering patterns for the cardiometabolic alterations. METHODS: One hundred people aged 19 to 77 were studied. Weight, height, abdominal circumference, blood pressure, glucose, lipid profile and insulin were all measured. Insulin resistance was estimated by HOMA. RESULTS: The five most prevalent cardiometabolic factors were low HDLc (90 %), overweight/obesity (78 %), abdominal obesity (68 %), hypercholesterolemia (59 %) and insulin resistance (54,8 %). Low HDLc frequency was higher in women (p <0,01). Subjects aged 35 to 55 demonstrated higher cardiometabolic factor frequency. Metabolic syndrome prevalence was 57 %, prevailing in women. Three or four factors were found in most individuals. The most commonly occurring factors in the identified clusters were overweight/obesity, hypercholesterolemia, abdominal obesity and low HDLc. The body mass index and insulin resistance predicted metabolic syndrome in the evaluated group. CONCLUSIONS: High cardiometabolic factor prevalence was found in the community being studied as well as a high tendency for several of these factors to cluster, thereby indicating the need to execute programmes focusing on reducing cardiometabolic risk. PMID- 20027512 TI - [Household costs of maintaining and/or abandoning exclusive breastfeeding during the first postpartum month]. AB - OBJECTIVE: Estimating the difference between direct and indirect family costs arising from women who continue exclusive breastfeeding and those who abandon it during the first postpartum month. METHODS: A cost analysis was carried out on primiparous women from Cali, Colombia; they were recruited during their immediate puerpery and follow-up during the first 30 postpartum days. Non-parametric Bootstrap techniques were used for estimating average costs and differences for 0 8, 9-30 and 0-30 postpartum days regarding women who abandoned exclusive breastfeeding and those who did not. RESULTS: Women who abandoned exclusive breastfeeding assumed higher costs related to baby feeding while those who continued exclusive breastfeeding had higher food costs for the mother and in contracting domestic support. CONCLUSION: The savings which a woman and her family can make due to continuing with exclusive breastfeeding become negated by the mother's food-related costs. Interventions should be designed which are orientated towards prolonging exclusive breastfeeding and modifying beliefs about nutrition which contribute towards generating economic consequences for families. PMID- 20027513 TI - [The gateway to illegal substance use in Colombia: violations of the gateway rule]. AB - OBJECTIVE: Establishing the frequency of gateway illegal substance use with alcohol or nicotine being violated amongst Colombian adults in the general population. METHODS: A descriptive study was carried out on adults from the general Colombian population. Gateway frequencies were determined for illegal substance use regarding substances other than alcohol or nicotine. RESULTS: A total of 4,426 adults participated in this research. A group of 127 people (3.3%) reported that they had started illegal substance use before using alcohol or nicotine; 2.3% of them had used other illegal substance before using cannabis, 0,6 % had smoked cannabis before using alcohol or nicotine and 0,4 % had used other illegal substances before using alcohol or nicotine. CONCLUSIONS: A small group of people who use illegal substances in Colombia start with substances different to alcohol or nicotine. The factors associated with gateway use of illegal substances (regarding substance other than alcohol or nicotine) should be investigated. PMID- 20027514 TI - [State school children's opinions regarding violence in the media]. AB - OBJECTIVE: To describe the opinion of schoolage children, from a public school, regarding the violence they perceive in the media. MATERIAL AND METHODS: Descriptive-exploratory research with a sample, selected according to the child's age in a public school. Quantitative data were collected. RESULTS: There were found common issues related to the child's opinion about the violence present in the media such as frequent exposure to the media violence and lack of parental supervision. CONCLUSIONS: Social context and constant exposure to the media violence affect the children's opinion about the violence phenomenon and their predisposition to it. PMID- 20027515 TI - [Estimating the morbidity profile amongst Colombian civil aviation personnel]. AB - OBJECTIVE: Describing main morbidity rate amongst Colombian civil aviation cockpit and air traffic control personnel. METHODS: This was a cross-sectional study. A random sample of 1,059 records (pilots, flight attendants, air traffic controllers) was taken from Colombian aeronautical authority files. Identification, background, morbidity and limitation data was reviewed for each one. RESULTS: 68,2 % of the records concerned males. Average age was 37. 22,9 % of the sample had some medical antecedent recorded on their first certification. Alcohol consumption (occasional) was recorded for 17,1 % of the sample and 11,9 % were smokers. The main diagnoses made were: vision disorders (52,2 %), dyslipidemias (40,6 %), being overweight (37,5 %), having hearing loss (22,0 %), being healthy (14,0 %) and suffering from hypertension (8,1 %). Each group had the same diagnoses, but occurring in a different order. CONCLUSIONS: Others studies have found cardiovascular disease to be the main cause of morbidity amongst cockpit and air traffic control personnel. Vision disorders were the main disorder occurring in Colombia. We do not know if there is a predisposition amongst Colombians to suffer such morbidity. Cardiovascular risk factors were found in second place in Colombia, meaning that public health prevention and promotion countermeasures should be taken. PMID- 20027516 TI - [A study of self-medication in a neighborhood in Bogota]. AB - OBJECTIVE: Establishing the prevalence of people living in the Suba district of Bogota using medication at their own risk and determining the factors related to it. METHODS: A guided survey was applied to a random sample. Demographic variables were measured and inquires were made about health problems related to self- medication, the reasons for not seeing a doctor and the medications consumed during the last two weeks. RESULTS: Self-medication reached 27,3 % (19,2 35,3 % 95 % CI) and self-prescription was 7,7 % (2,8-12,5 % 95 % CI). Being affiliated to the Social Health Security's beneficiaries system (OR=2,61: 1,4-4,8 95 % CI) was related to such behaviour. No relationship with other variables was found. The medications most consumed by people indulging in self-medication were analgesics (59,3 %), anti-flu medicine (13,5 %) and vitamins (6,8 %). The main problems for which people resorted to self-medication were pain, fever and flu. The main reasons mentioned for not seeing a doctor were lack of time (40 %) and lack of economic resources (43 %), in addition to other arguments, such as the people s perception that the problem was mild and emergency rooms are always congested. CONCLUSIONS: Although still worrying from the public health point of view, self-medication figures were lower than those encountered in similar studies; self-medicated drugs were the over-the-counter medication type. The rate of consuming self-medicated antibiotics dropped, probably due to the spreading of the restriction on the sale of antibiotics without a medical prescription which came into force a few months before the survey took place. PMID- 20027517 TI - [Temporal prediction of the dengue epidemic in Colombia: the epidemic's probabilistic dynamics]. AB - OBJECTIVE: Studying geometric dynamics for annual cases of dengue recorded in Colombia from 19902006 by making an analogy with a probabilistic random walk. METHODS: The geometric dynamics for the number of annual cases of dengue registered in Colombia for 1990-2006 was mathematically analyzed as a probabilistic random walk, building the total probability space for the dynamics in order to analyze the probabilistic behavior of consecutive increases and decreases and the probabilistic behavior of cases during ranges of serial years and thus calculate the temporary prediction of annual cases. RESULTS: The extreme values predicted for the number of infected people during 2007 were 57 581 and 20 008; the predicted value was refined by analysing annual variations, giving a 48 188 predicted value for the number of infected people in 2007. This prediction was then corroborated by using Colombian National Institute of Health data having a 43 564 recorded real infected value on December 31st; the real value corresponded to 90,4 % regarding predicted value. CONCLUSION: A mathematical and acausal order for the dengue epidemic in Colombia was found from the probabilistic random walk; thus led to carrying out temporary, simple and useful predictions in the public health decision-making. PMID- 20027518 TI - [Evaluating the burden of disease caused by human papillomavirus in Bogota]. AB - OBJECTIVE: A study was carried out in Bogota aimed at estimating the burden of disease associated with human papillomavirus infection (HPV) and the potentially avoidable percentage due to using new vaccines. METHODS: A literature review was combined with analysing surveillance system data and disease cost evaluation. RESULTS: After adjusting for underreporting and misclassification, it was estimated that 322 deaths from cervical cancer occur annually in Bogota (corresponding to 676 new cases). This would cause the loss of 15 years of life for each 1,000 women per year (most occurring amongst women aged 40 to 69). In addition to cervical cancer, there would be around 6,084 cases of high-grade and 22,984 low-grade cervical lesions yearly. The disease's yearly cost would amount to around 7 million dollars. Important weaknesses were found in the clinical management of women suffering from cervical lesions. CONCLUSION: Strengthening cervical cancer prevention programmes in Bogota would lead to saving a significant number of deaths, cases of cancer and the costs associated with HPV infection. Introducing an anti-HPV vaccine may be considered, but only as part of a more widespread preventative strategy and provided that more affordable prices have been found. PMID- 20027519 TI - [Sexually-transmitted disease prevalence and the factors associated with it in sexually-exploited children in Bogota, Colombia]. AB - OBJECTIVE: Presenting the characteristics of a group of children who have been sexually exploited in Bogota, determining the prevalence of sexually-transmitted disease (STD) in this group of children and establishing the factors associated with STD in this population. METHODOLOGY: This was a descriptive, cross-sectional study. The medical charts of children aged 10 to 17 years who sought medical attention at Fundacion Renacer between January 2002 and June 2004 were reviewed. The analysis included descriptive statistics, correlations and binomial logistic regression. RESULTS: 255 children's charts were included, producing the following characteristics: being female (58.8%), average age was 15.4, 5.9% were affiliated to social security, 89.8% engaged in substance abuse, 41.2% used family planning, 3.9% used a condom during every intercourse and there were 0.86 average pregnancies per female adolescent. Sexually-transmitted disease prevalence was 30.9%. The most common diseases were condilomatosis and Neisseria gonorrhoeae infection. Substance abuse and infrequent condom use were associated with a diagnosis of STD. CONCLUSION: Child sexual exploitation is a reality in Colombia. In spite of the limitations of this study, it can be concluded that STDs are an important problem in the sexually-abused child population in Bogota. Preventing child sexual exploitation and integral attention for these children (including health service provision) must become a priority. PMID- 20027520 TI - [Seroprevalence of Lyme borreliosis in workers from Cordoba, Colombia]. AB - OBJECTIVE: Establishing the seroprevalence of Lyme disease in workers from Cordoba. MATERIAL AND METHODS: Representative serum samples (152) were taken from the cities of Monteria, Cerete, Lorica and Cotorra in the Cordoba department in Colombia. Borrelia burgdorferi antibodies were detected by ELISA and confirmed by western blot anti-Borrelia blot assay. RPR (rapid plasma regain) test for Treponema pallidum (syphilis) and ELISA for leptospirosis IgM were carried out to discard cross-reactivity. RESULTS: A total of 152 serum samples were tested; 30 (20 %) were positive by ELISA. The 30 positive sera were RPR negative. Seven sera were confirmed by western blot; seroprevalence was 4.6 %. Reactivity against p41, p58, p75, OspA, p30, OspC, p17, VLsE and p83/100 were detected. CONCLUSIONS: Our results revealed antigenic evidence of Borrelia in the rural area of Cordoba. Identifying clinical, ecological and serological cases, linked to searching for Borrelia burgdorferi in infected tick vectors must be carried out in rural parts of Colombia. PMID- 20027533 TI - [Electric stimulation in dysphagia therapy--a review]. AB - In the last years an increased interest in the electrical stimulation has consisted in the treatment of dysphagia. In the article we introduce the anatomical and physiological premises for the method. In a critical analysis the present state of art is represented, the clinical results are checked and the chances for the future are examined. PMID- 20027535 TI - [The relevance of anatomical courses in ENT-education]. AB - BACKGROUND: Dissection-courses have become an important element in ENT-education. They are offered covering different subjects. Until now no study exists that verifies the acceptance and efficiency of those courses. METHODS: We consulted the trainees of three surgical anatomical courses with a multiple choice form for their opinion on courses visited in the past. The acquired data was analyzed quantitatively and examined for statistically significant correlation between age, gender and professional experience. RESULTS: In our study 73 participants filled in our questionnaire. The average age was 32 years. The satisfaction concerning the currently offered courses is high. The dissection-exercises were rated as 'good' and 'very good' by 72%. The strongest point of criticism (49%) related to the price of the dissection-courses. The improvement of surgical skills showed to be the main motivation for visiting anatomical dissection courses (46%). Most respondents had registered for dissection-courses of the temporal bone and the paranasal sinuses. A correlation between professional experience and the type of courses visited could be found for the courses on nerval- and blood vessel reconstruction. DISCUSSION: The courses offered currently in ENT are appreciated as beneficial and effective. The fact that these courses are visited frequently and that the benefit in daily practice is highly appreciated indicate a well thought out educational concept. It seems unlikely that fees for these courses can be reduced as hoped for by many participants. The economic pressure on the organizational team is very high. Surprisingly the interest in CME-points is relatively low. PMID- 20027536 TI - [Intention of the patient certain the surgeons election]. PMID- 20027537 TI - [Acupuncture in the HNO-area]. AB - For ENT specialists it is good to become familiar with the basics of acupuncture, as in our times this method of treatment has been spread and accepted worldwide. Modern pain research has succeeded in scientific exploring and verifying acupuncture effects. Any medical specialist having understood the facts and implications of acupuncture as laid out in this paper, may take the chance to try and put acupuncture into practice in his/her daily work. One of the challenges of doctors nowadays is the increase of functional disorders, of myofascial pain syndromes, of patients' general ill-feeling of health. Particularly in pain management, as regards to the considerable toxic by-effects of long-term prescribed analgetics, there is a need of gentle therapies such as acupuncture, free from side-effects. It is the patients who demand alternative concepts. For ENT-doctors running a busy office, microsystem acupuncture in particular may be recommended. Inserting a couple of needles at the auricle, or injecting a few drops into the oral mucosa is a quick action which in most cases will provide beneficial results. PMID- 20027538 TI - Validity of the Nike+ device during walking and running. AB - We determined the validity of the Nike+ device for estimating speed, distance, and energy expenditure (EE) during walking and running. Twenty trained individuals performed a maximal oxygen uptake test and underwent anthropometric and body composition testing. Each participant was outfitted with a Nike+ sensor inserted into the shoe and an Apple iPod nano. They performed eight 6-min stages on the treadmill, including level walking at 55, 82, and 107 m x min(-1), inclined walking (82 m x min(-1)) at 5 and 10% grades, and level running at 134, 161, and 188 m x min(-1). Speed was measured using a tachometer and EE was measured by indirect calorimetry. Results showed that the Nike+ device overestimated the speed of level walking at 55 m x min(-1) by 20%, underestimated the speed of level walking at 107 m x min(-1) by 12%, but closely estimated the speed of level walking at 82 m x min(-1), and level running at all speeds (p<0.05). Similar results were found for distance. The Nike+ device overestimated the EE of level walking by 18-37%, but closely estimated the EE of level running (p<0.05). In conclusion the Nike+ in-shoe device provided reasonable estimates of speed and distance during level running at the three speeds tested in this study. However, it overestimated EE during level walking and it did not detect the increased cost of inclined locomotion. PMID- 20027539 TI - A 3-year longitudinal analysis of changes in Body Mass Index. AB - The aim of this study was to analyse whether Physical Activity Index (PAI), Physical Fitness, Screen Time (watching TV and computer use), Socio-economic Status and Commuting to School made a significant contribution to longitudinal changes in Body Mass Index (BMI) in youth. This longitudinal study was carried out over a period of 3 years with 345 students (147 boys) who were between 11 and 16 years old at the beginning of the study. Students were invited to perform tests from FITNESSGRAM Battery for Curl-Ups, Push-Ups, Back-Saver Sit and Reach, and 20 m Shuttle-Run (CRF). Fitness tests were categorized in "Healthy Zone" (HZ) and "Under Healthy Zone" (UHZ), PAI in "less active" and "active"; Socio-economic Status, in low, middle and high education level, and Commuting in active and passive. BMI was corrected for age and gender meaning that we subtracted the age and-sex-specific cut points for overweight. Corrected body mass index was used as dependent variable in a Linear Mixed Model. The main result was the strong positive and independent association of individuals with CRF performances UHZ with corrected body mass index. In conclusion, the results of this longitudinal study showed markedly an important relationship of lower fitness levels with the risk of being overweight/obese, in particular CRF and abdominal strength. PMID- 20027540 TI - Lumbar arachnoiditis and thecaloscopy: brief review and proposed treatment algorithm. AB - The term arachnoiditis describes the inflammation of the meninges and subarachnoid spaces. Lumbar arachnoiditis is characterized by obliterated nerve root sleeves and the adherence of nerve roots to each other in the proximity of the cauda equina, and may be secondary to infectious diseases or tumors, iatrogenic (subsequent to spinal surgery) or idiopathic. It is not very clearly defined epidemiologically or clinically, and various theories regarding its pathophysiology have been proposed; furthermore, its treatment is difficult because there is a lack of evidence-based diagnostic and therapeutic gold standards. Thecaloscopy has been recently described as a novel technique for retrograde transcutaneous neuroendoscopic inspection of the subarachnoid structures of the lumbar thecal sac; it has also been suggested for the treatment of lumbar arachnoiditis. We here review the most modern techniques for the treatment of this disease such as thecaloscopy and neurostimulation. PMID- 20027541 TI - Intraoperative near-infrared fluorescence imaging in perforator flap reconstruction: current research and early clinical experience. AB - Despite recent advances in perforator flap reconstruction, there can be significant variability in vessel size and location. Although preoperative evaluation may provide valuable information, real-time intraoperative methods have the potential to provide the greatest benefit. Our laboratory has developed the Fluorescence-Assisted Resection and Exploration (FLARE) near-infrared (NIR) fluorescence imaging system for intraoperative visualization of details of the underlying vasculature. The FLARE system uses indocyanine green, a safe and reliable NIR fluorophore already FDA-approved for other indications. The system has been optimized in large-animal models for the identification of perforator size, location, and perfusion and has also been translated to the clinic for use during breast reconstruction after mastectomy. In this article, we review our preclinical and clinical data, as well as literature describing the use of similar NIR fluorescence imaging systems in plastic and reconstructive surgery. PMID- 20027542 TI - Colonic atresia: a clinicopathological insight into its etiology. AB - BACKGROUND: The pathogenesis of colonic atresia (CA) has not been discussed in detail, although the theory of a prenatal vascular insult, hypothesized from patients with small bowel atresia, has been accepted in the past. This review questions this etiology by reviewing all patients with CA treated in two institutions. MATERIAL AND METHODS: A retrospective analysis was done of the medical notes of 30 patients with CA treated in two tertiary centers of neonatal surgery in Glasgow, UK, and in Wroclaw, Poland, over a 30-year period with special emphasis on the intraoperative findings and morphology of the atretic bowel. RESULTS: Thirteen patients had CA in association with an abdominal wall defect. Eleven patients had gastroschisis. A single fibrous cord atresia was noted in 2 patients and it was located in the ascending colon and transverse colon, respectively. Four patients had type IIIa atresia affecting the ascending colon in 3 and transverse colon in one. In 5 patients an extensive defect of the intestine with two atresias of the small and large bowel was noted. In 3 of them, a separate conglomerate of the prolapsed intestine was found to be necrotic or presenting as a cystic structure. Two of these patients had an unusually narrow abdominal wall defect of less than 1 cm. Isolated CA was noted in 17 patients. A type IIIa atresia affecting the right colon was found in 14 of them. In 2 patients the ascending colonic atresia was accompanied by an extensive defect of the transverse colon. A single sigmoid colon atresia and two fibrous cord atresias of the sigmoid and transverse colon were noted in single patients respectively. CONCLUSIONS: Most cases of CA in babies with gastroschisis seem to result from bowel compression within the narrowing abdominal defect. A "two-point constriction" may lead to a wide spectrum of bowel pathologies and the morphology may depend on the viability of the intestinal segment between the atretic jejunum or ileum and the colon. Isolated CA presents with a wider spectrum in terms of anatomical types and location of the bowel pathology, but it seems that type II and IIIa atresias of the right colon may share a similar pathogenesis of temporary constriction within a closing umbilical ring. PMID- 20027543 TI - [Tumor surgery--what lies ahead?]. PMID- 20027544 TI - [Decision of the Cologne Federal Court November 6 2008, section O 410/06- compartment syndrome: failure to recognize and insufficient treatment in childhood after operation]. PMID- 20027545 TI - Intracranial calcifications on CT. AB - Computed tomography (CT) is very sensitive for detection and localization of intracranial calcifications. We reviewed in this pictorial essay the diseases associated with intracranial calcifications and emphasized the utility of CT for the differential diagnosis. PMID- 20027546 TI - Pancreatic metastases: CT and MRI findings. AB - PURPOSE: The pancreas is an unusual but occasionally favored site for metastases in patients with advanced malignancy. The pivotal role of computed tomography (CT) and magnetic resonance imaging (MRI) examination in detecting the pancreatic mass and providing guidance to obtain a definitive tissue diagnosis is emphasized in this study. MATERIALS AND METHODS: Eleven patients with pancreatic metastases, 4 women and 7 men with a mean age of 62.45 years, were examined in a period of 5 years. All patients underwent CT examination and 3 patients were further evaluated by MRI. RESULTS: The primary malignancy was lung carcinoma in 7 patients, breast carcinoma in 3 patients and renal cell carcinoma in 1 patient. The pancreatic metastases were detected during initial staging in 4 cases and during follow-up in 7 cases. The type of metastases was solitary in 7 patients, multiple in 3 patients and diffuse in 1 patient, with rim or homogeneous enhancement. Seven patients had metastases to other organs. CT-guided biopsy was performed in 5 patients. CONCLUSION: Disparity in prognosis and management of patients with secondary pancreatic tumors makes detection and characterization of metastases to the pancreas an important goal of CT and MRI evaluation. PMID- 20027547 TI - Dual-echo TFE MRI for the assessment of myocardial iron overload in beta thalassemia major patients. AB - PURPOSE: Cardiac failure due to myocardial iron overload is the most common cause of death in beta-thalassemia patients. Multi/ two echo times-turbo field echo (TE TFE) magnetic resonance imaging (MRI) is considered the gold standard technique in the evaluation of myocardial iron accumulation. However, multi TE-TFE technique is not available in all scanners. The aim of our study was to show the role of black blood dualecho cardiac triggered TFE in the assessment of myocardial iron overload. MATERIALS AND METHODS: Sixteen beta-thalassemia major patients (10 males) with a mean age of 19 years who were receiving parenteral deferoxamine and oral deferiprone treatment were included in this study. Baseline measurement of myocardial T2* values were < 20 ms in all patients. Cardiac MRI was performed after 6 months, 12 months, and 18 months with the same technique. RESULTS: The average baseline value of T2* was 8.2 +/- 3.6 ms. After treatment of combined deferoxamine and deferiprone, the average measurements of myocardial T2* at 6, 12, and 18 months were 11.3 +/- 6.0, 13.6 +/- 7.5, and 15.7 +/- 7.4 ms, respectively (P < 0.05). The basal ejection fraction (EF) value was 49 +/- 8.7%. The EFs were 54.4 +/- 11% at 6 months, 54.8 +/- 6.9% at 12 months, and 58.6 +/- 3.6% at 18 months of followup (P > 0.05). CONCLUSION: Cardiac MRI with dual TE TFE technique can be used to determine myocardial iron accumulation and response to the chelation treatment. PMID- 20027548 TI - Fast MRI evaluation of pulmonary progressive massive fibrosis with VIBE and HASTE sequences: comparison with CT. AB - PURPOSE: The aim of this prospective study was to evaluate the diagnostic utility of volumetric interpolated breath-hold examination (VIBE) and half-Fourier acquisition single-shot turbo spin-echo (HASTE) fast magnetic resonance imaging (MRI) sequences in the evaluation of pulmonary progressive massive fibrosis (PMF) in comparison with computed tomography (CT) imaging. If fast MRI is proven to be diagnostically significant, this modality can be used for diagnosis and follow-up studies of PMF patients. MATERIALS AND METHODS: Twenty-two PMF lesions from 20 coal workers were evaluated. After CT imaging, patients underwent pre-contrast VIBE, contrast-enhanced VIBE, and HASTE MRI studies for detection and evaluation of the PMF lesions. Measurements of the three groups were evaluated with intra class coefficients. Correlation levels between sizes, image quality, and artifact were evaluated with linear Pearson correlation analysis. RESULTS: There was almost perfect agreement among radiologists for lesion detection with kappa analysis. There was significant agreement between three MRI study groups and gold standard CT images. We found the best agreement values with contrast- enhanced VIBE images for lesion detection and image quality in comparison with CT imaging. Presence of artifact was also lowest with this protocol. CONCLUSION: With fast MRI sequences in pulmonary imaging, image quality has significantly improved being very close to that of CT studies. In this study, contrast-enhanced VIBE protocol provided the best depiction of PMF lesions. This protocol may be an alternative choice for CT, avoiding the use of iodinated contrast material and minimizing exposure to ionizing radiation for follow-up studies. PMID- 20027549 TI - MR tractography of schizencephaly. AB - This report attempts to document the potential of magnetic resonance (MR) tractography in providing detailed information on white matter tract anomalies in schizencephaly. Diffusion tensor imaging with tractography was performed in four patients referred for neuroimaging. On conventional MR imaging, two of the patients had temporal lobe involvement and the other two had frontal lobe involvement. Tractography was successful in showing major tract abnormalities in two patients and subcortical involvement in three. Tractography was negative in one patient. MR tractography is a promising imaging method in developmental anomalies, beneficial for both clinical management and understanding the nature of the pathology. PMID- 20027550 TI - Management of benign prostatic hypertrophy-related urinary retention: current trends and perspectives. AB - The failure of prostate-directed treatment modalities to help all men or all symptoms has questioned the longstanding assumption that the prostate is at the root of all male urinary symptoms, and a correlation between urinary bladder function and prostate pathology has been recognized. Now, it is widely recognized that bladder dysfunction plays a role in some, if not most, of the benign prostatic hyperplasia-related symptoms and signs, and recent studies have suggested that pharmacotherapies that target the bladder, such as antimuscarinics, may improve storage urinary symptoms. Indeed, the current mainstays of overactive bladder syndrome pharmacotherapy are antimuscarinic agents with mixed actions, including musculotropic (calcium antagonistic) activity. Moreover, the combination therapy with alpha blocker and antimuscarinic agents is now suggested when bladder outlet obstruction related to benign prostatic hyperplasia coexists with overactive bladder symptoms. Combinational treatment, targeting to both decrease resistance to urine outflow through the prostatic urethra and increase bladder smooth muscle, may improve the bladder outlet surgery success rate, and perhaps it might reinforce the need to offer an additional trial without catheter in patients with benign prostatic hyperplasia who are likely to suffer from detrusor hypocontractivity. Currently, no clinical trial supporting the use of parasympathomimetic drugs in those with poor voiding and longstanding symptomatic benign prostatic hyperplasia exists in the literature; however, experimental studies present promising results. PMID- 20027551 TI - Relevance of levels of evidence to the urologist. PMID- 20027552 TI - Sterile water versus isotonic saline solution as irrigation fluid in percutaneous nephrolithotomy. AB - INTRODUCTION: We evaluated the safety of sterile water as an irrigation solution for percutaneous nephrolithotomy (PCNL). MATERIALS AND METHODS: Forty-four patients with kidney calculi were enrolled in this study and randomly divided into two groups for PCNL. Approaches to the calculi were through a single subcostal access with an Amplatz sheath, and either sterile water or isotonic saline solution was used as the irrigation fluid. Serum hemoglobin, haptoglobin, sodium, potassium, and creatinine were measured before and 12 hours after the procedure. The patients were evaluated for signs of transurethral resection of the prostate syndrome during the operation for 24 hours afterwards. RESULTS: The mean calculus size, irrigation volume, irrigation time, and age were not significantly different between the two groups. Hemolysis occurred in 10 and 9 patients in sterile water and saline groups, respectively. The mean change in haptoglobin level was -1.7 +/- 59 mg/dL in the sterile water and 11 +/- 55 mg/dL in the saline group. Also, the mean change in plasma sodium level was -2.2 +/- 4.7 and -0.4 +/- 3.8 in sterile water and saline groups, respectively. None of these values were significantly different between the two groups, nor were other laboratory values. None of the patients developed transurethral resection of the prostate syndrome or needed transfusion. CONCLUSION: Sterile water is an inexpensive alternative to isotonic saline for irrigation during PCNL. We did not find any difference between the two irrigation solutions regarding the safety; however, this should be confirmed further, especially for larger calculi. PMID- 20027553 TI - Laparoscopic management of adrenal lesions larger than 5 cm in diameter. AB - INTRODUCTION: Laparoscopic adrenalectomy remains a controversial procedure for large tumors. The incidence of adrenocortical carcinoma increases and technical difficulty of adrenalectomy increases as the size increases. We examined the outcome and complications of laparoscopic adrenalectomy for such lesions. MATERIALS AND METHODS: Twenty-nine patients underwent laparoscopic adrenalectomy, of whom 19 had tumors larger than 5 cm in diameter, having a median tumor size of 7.0 cm. They were compared with patients whose adrenal tumors were smaller than 5 cm. RESULTS: Patients with small tumors (< 5 cm) had a significantly shorter median operative time of 90 minutes as compared to 145 minutes in those with large tumors (> 5 cm). There was no significant difference in the median hemoglobin drop (1.05 g/dL versus 1.30 g/dL), time for starting oral intake (24 hours in both groups) or hospital stay (3.5 days versus 4.0 days) between patients with small and large tumors, respectively. There were no intra-operative complications except for 1 incidence of supraventricular tachycardia in a patient with a large pheochromocytoma. There were no major complications seen in any of the patients and no open conversions. Histopathology of large tumors revealed 16 benign tumors (8 pheochromocytomas, 4 adenomas, 2 ganglioneuromas, 1 pseudocyst, and 1 myelolipoma) and 3 malignancies, of which 1 was primary adrenocortical carcinoma and 2 were metastatic renal cell carcinoma. CONCLUSION: In experienced hands, laparoscopic adrenalectomy is safe and feasible for large functioning adrenal tumors. Large adrenal tumors suspicious of harboring malignancy with no peri-adrenal involvement can be tackled laparoscopically. PMID- 20027554 TI - Kidney transplant anastomosis: internal or external iliac artery? AB - INTRODUCTION: We compared the best technique for arterial anastomosis in kidney transplantation, end-to-side anastomosis to the external iliac artery or end-to end anastomosis to the internal iliac artery. MATERIALS AND METHODS: A total of 38 patients with end-stage renal disease who received a kidney transplant from a deceased donor were randomized into two groups in order to undergo either end-to end anastomosis to the internal iliac artery or end-to-side anastomosis to the external iliac artery. Length of arterial anastomosis, cold ischemia time, hospital stay, serum creatinine level, recovery of urinary output, and surgical and clinical complications during hospitalization were evaluated. After 3 years, in the patients with a functioning allograft, creatinine clearance measure, Doppler ultrasonographic study, survival, graft loss, and erectile function were compared between the two groups. RESULTS: Postoperative analyses showed similar recovery of urinary output (P = .39) and creatinine (P = .95) between the two groups. No differences in clinical (P = .55) and surgical (P = .80) complications or in hospital stay (P = .90) were noted. The 3-year follow-up demonstrated no differences in Doppler ultrasonography results, creatinine clearance (P = .80), patient survival (P = .22), and graft loss (P = .72). Erectile dysfunction was similar, being related only to pre-operative medical history and age. CONCLUSION: Both techniques showed similar results in short- and long-term follow-ups. Larger prospective studies are warranted to clarify the risk of renal artery stenosis and development of erectile dysfunction. PMID- 20027555 TI - Sildenafil or vardenafil nonresponders' erectile response to tadalafil. AB - INTRODUCTION: Erectile dysfunction has usually been treated by a phosphodiesterase 5 inhibitor in men, especially in the past decade. Although sildenafil and vardenafil are widely used, there is a high percentage of people who do not respond to these drugs. This study was performed in order to evaluate the efficacy of the lastly presented phosphodiesterase 5 inhibitor, tadalafil, in nonresponder group of patients to sildenafil and vardenafil. MATERIALS AND METHODS: Forty married men with erectile dysfunction who had taken sildenafil or vardenafil at the maximum recommended doses and had not responded to the treatment were included. They were treated with tadalafil, 20 mg, at least 4 doses at different days. The effectiveness of the treatment was reviewed by different questionnaires, including the International Index of Erectile Function 5 (IIEF-5), Sexual Encounter Profile (SEP) questions 2 and 3, and the Global Assessment Question (GAQ), at the end of the 12th week. RESULTS: The IIEF-5 scores were 11.90 +/- 4.78 and 12.67+/-6.70, before and after at least 4 doses of tadalafil, respectively (P = .30). The rate of positive responses to SEP2, SEP3, and GAQ questions were also insignificantly different after the treatment. During this period, flushing was seen in 10 and headache was seen in 5 patients. CONCLUSION: The recommended maximum dose for tadalafil insignificantly improved the IIEF5, SEP2, SEP3, and GAQ scores in patients with erectile dysfunction who had not responded to sildenafil and vardenafil. The other treatment alternatives should be in mind after getting no response to the optimum doses and enough trials of sildenafil or vardenafil before trying a tadalafil regimen. PMID- 20027556 TI - Delayed retropubic urethroplasty of completely transected urethra associated with pelvic fracture in girls. AB - INTRODUCTION: The objective of the present study was to evaluate the results and the complications of delayed retropubic urethroplasty of completely transected urethra associated with pelvic fracture in girls. MATERIALS AND METHODS: From 2002 to 2008, a total of 7 girls with complete urethral disruption after pelvic fracture were referred to our center and all of them underwent delayed retropubic urethroplasty with end-to-end anastomosis of the urethra. RESULTS: Seven female patients with a median age of 6 years old underwent delayed end-to-end anastomosis. The median time to surgery was 6 months from the trauma. Voiding was normal after catheter removal in all of the patients. The median follow-up was 36 months. Three patients had mild stress urinary incontinence after catheter removal. CONCLUSION: There are some different strategies for management of complete urethral avulsion in females who have sustained pelvic fracture, including early realignment, bladder flaps, and end-to-end anastomosis. The strategy of delayed end-to-end anastomosis urethroplasty with retropubic approach is sound and produces acceptable results. The use of flexible cystoscope and omental flap is effective in achieving continence after urethroplasty in such cases. PMID- 20027557 TI - The use of unaltered appendix transfer in ileal continent reservoir: 10 years experience, a novel technical modification. AB - INTRODUCTION: We report a new modified technique of unaltered appendix transfer to ileal pouch and preserving ileocecal segment. This modification enables us to use ileum as the popular type of enteric segment instead of ileocecal segment while using appendix as a catheterizable stoma. MATERIALS AND METHODS: Forty-five patients (30 men) who needed reconstruction of the lower urinary tract were enrolled for using appendix as a catheterizable stoma. Reservoir was reconstructed using ileal segment. The appendix was circumcised from its base over its pedicle. The spatulated appendix tip was exteriorized as a catheterizable stoma to the skin, preferably umbilicus, and its base was implanted to the ileal pouch. RESULTS: Follow-up records of 38 of 45 patients were available. The median follow-up period was 29 months. The mean intermittent catheterization interval was 4.19 +/- 1.6 hours. Urodynamic parameters were evaluated for 18 out of 38 patients. The median maximal pouch capacity determined as 380 mL. The median appendiceal closure pressure was 61 cm H2O. No pouch perforation occurred. Stomal stenosis occurred in 3 patients. They did not catheterize their appendiceal stoma because they restarted catheterization through the urethra. CONCLUSION: This novel approach enabled us to use ileum as today's more popular type of bowel segment to reconstruct enteric pouch rather than using ileocecal segment, while using appendix as a catheterizable stoma. One of the unique advantages of this technique is that the postponement of clean intermittent catheterization will not result in pouch perforation since the urine will leak when the pouch becomes overfill. PMID- 20027558 TI - Special characteristics of culturing mature human bladder smooth muscle cells on human amniotic membrane as a suitable matrix. AB - INTRODUCTION: Our aim was to evaluate the natural behavior, growth pattern, morphology, and specific features of human bladder smooth muscle cells (HBSMCs) on two different matrixes, including human amniotic membrane (HAM) and collagen. MATERIALS AND METHODS: The HBSMCs were obtained from 6 children with primary vesicoureteral reflux undergoing open antireflux surgery, and they were isolated from the anterior wall of the bladder. The specimens were cultured on a tissue culture plate of bovine dermal collagen serving as control and on decellularized HAM. Histological, transmission electron microscopy, and immunocytochemical examinations were done, thereafter. RESULTS: On HAM, very few HBSMCs slowly migrated from explant tissue on the 7th day of culture. All the cells were placed at the same direction, and in some parts, formed multilayer. After 35 to 40 days, the confluency rate was 75% and the cells were orderly arranged. On collagen, cell migration from explant culture took place as rapidly as the 3rd to 4th day of culturing. On days 30 to 40, the confluency rate was 100%. Immunocytochemical staining was positive for anti-actin and antidesmin antibodies. On transmission electron microscopy, cell organelles of HBSMCs exhibited the same features of the natural smooth muscle cells. They were tightly attached to each other and the underlying layer basement membrane. CONCLUSION: A well-designed growth pattern of HBSMCs on HAM with abundant cell-to-cell adhesions encourages us to use it as a competent tissue for reconstruction of relatively damaged or diseased bladders. Undoubtedly, further clinical studies should be performed to replicate our results. PMID- 20027559 TI - Inductive role of collagen type IV during nephrogenesis in mice. AB - INTRODUCTION: During nephrogenesis, transition of mesenchyme to the epithelium of tubules and glomeruli occurs via the interaction of ureteral bud and metanephric mesenchyme. The distribution pattern of collagen type IV suggests that a regulated balance of activities is required to facilitate migration of the ureteral bud branches into the mesenchyme and to control early extracellular matrix changes during tubulogenesis. We used a specific antibody for tracing collagen type IV basement membrane during renal tubules morphogenesis. MATERIALS AND METHODS: Twenty female Balb/C mice were divided randomly into 10 groups and were kept until finding vaginal plug was as an indicator of day zero of pregnancy. Twelve pregnant mice were sacrified by cervical dislocation in one of gestational days 13 to 18 and their fetuses were fixed, serially sectioned, and underwent immunohistochemical study for tracing of collagen type IV in basement membrane of glomeruli. The same processes were used for kidneys preparation on postnatal days 5, 10, 15, and 20 in newborns of 2 mothers for each day. RESULTS: Collagen type IV showed weak reaction on day 14 of gestation in tubular basement membrane. The amount of collagen increased continuously until the following days of fetal life and of the first 5 postnatal days in basement membrane. After this period, collagen type IV reaction was not showed significant change in newborns. CONCLUSION: These results indicate that developmental changes in various nephron segments from most immature stages to most differentiated structures are dependent on the collagen type IV expression. PMID- 20027560 TI - A safe surgical approach to a giant intrarenal arteriovenous fistula and aneurysm. PMID- 20027561 TI - Persistent mullerian duct syndrome with an irreducible inguinal hernia. PMID- 20027562 TI - Fibroepithelial congenital polyp of prostatic urethra in an adult man. PMID- 20027563 TI - Extramammary Paget's disease in prostate. PMID- 20027564 TI - Successful macrosurgical reimplantation of an amputated penis. PMID- 20027567 TI - Fat and water magnetic resonance imaging. AB - A wide variety of fat suppression and water-fat separation methods are used to suppress fat signal and improve visualization of abnormalities. This article reviews the most commonly used techniques for fat suppression and fat-water imaging including 1) chemically selective fat suppression pulses "FAT-SAT"; 2) spatial-spectral pulses (water excitation); 3) short inversion time (TI) inversion recovery (STIR) imaging; 4) chemical shift based water-fat separation methods; and finally 5) fat suppression and balanced steady-state free precession (SSFP) sequences. The basic physical background of these techniques including their specific advantages and disadvantages is given and related to clinical applications. This enables the reader to understand the reasons why some fat suppression methods work better than others in specific clinical settings. PMID- 20027568 TI - Short echo time 1H MRSI of the human brain at 3T with adiabatic slice-selective refocusing pulses; reproducibility and variance in a dual center setting. AB - PURPOSE: To assess the reproducibility of (1)H-MR spectroscopic imaging (MRSI) of the human brain at 3T with volume selection by a double spin echo sequence for localization with adiabatic refocusing pulses (semi-LASER). MATERIALS AND METHODS: Twenty volunteers in two different institutions were measured twice with the same pulse sequence at an echo time of 30 msec. Magnetic resonance (MR) spectra were analyzed with LCModel with a simulated basis set including an experimentally acquired macromolecular signal profile. For specific regions in the brain mean metabolite levels, within and between subject variance, and the coefficient of variation (CoV) were calculated (for taurine, glutamate, total N acetylaspartate, total creatine, total choline, myo-inositol + glycine, and glutamate + glutamine). RESULTS: Repeated measurements showed no significant differences with a paired t-test and a high reproducibility (CoV ranging from 3% 30% throughout the selected volume). Mean metabolite levels and CoV obtained in similar regions in the brain did not differ significantly between two contributing institutions. The major source of differences between different measurements was identified to be the between-subject variations in the volunteers. CONCLUSION: We conclude that semi-LASER (1)H-MRSI at 3T is an adequate method to obtain quantitative and reproducible measures of metabolite levels over large parts of the brain, applicable across multiple centers. PMID- 20027569 TI - Performance of imaging modalities in diagnosis of liver metastases from colorectal cancer: a systematic review and meta-analysis. AB - Surgery of liver metastases can be effective, and the appropriate selection of surgical candidates relies first on imaging. Different techniques are available, but information on their relative performance is unclear. The aim of this overview is to assess the imaging modality performance in the diagnosis of colorectal cancer (CRC) liver metastases. MEDLINE and EMBASE were searched for articles published from January 2000 to August 2008. Eligible trials had to be conducted on patients with diagnosis/suspicion of CRC liver metastases, comparing more than two modalities among MRI, computed tomography (CT), positron emission tomography using fluoro-18-deoxyglucose (FDG-PET), ultrasonography (US). Pooled estimates of sensitivity, specificity were calculated and pair-wise comparisons were performed. Of 6030 screened articles, 25 were eligible. Sensitivity and specificity on a per-patient basis for US, CT, MRI, and FDG-PET were 63.0% and 97.6%, 74.8% and 95.6%, 81.1% and 97.2, and 93.8% and 98.7%, respectively. On a per-lesion basis, sensitivity was 86.3%, 82.6%, 86.3%, and 86.0%, respectively. Specificity was reported in few studies. MRI showed a better sensitivity than CT in per-patient (odds ratio [OR]: 0.69; 95% confidence interval [CI]: 0.47-0.99; P = 0.05) and in per-lesion analysis (OR: 0.66; 95% CI: 0.55-0.80; P < 0.0001). In per-lesion analysis, the difference was higher when liver-specific contrast agents were administered. Available evidence supports the MRI use for the detection of CRC liver metastases. PMID- 20027570 TI - Detection of normal spinal veins by using susceptibility-weighted imaging. AB - PURPOSE: To evaluate the visualization of the spinal veins using susceptibility weighted imaging (SWI). MATERIALS AND METHODS: A 1.5-T magnet equipped with a spine matrix coil was used. Axial SWI scans of 20 healthy volunteers were obtained with a three-dimensional fast low-angle shot (3D-FLASH) sequence. Maximum intensity projection (MIP) of the phase images were reconstructed and five MIP images (at the levels of T11, T11/12, T12, T12/L1, and L1) were selected for the evaluation. The anterior median vein (AMV), posterior median vein (PMV), anterior radiculomedullary vein (ARV), posterior radiculomedullary vein (PRV), and sulcal vein (SV) were evaluated using a 4-grade scale (0, none; 1, weak; 2, moderate; and 3, prominent). RESULTS: The AMV was detected in all the subjects (100%). The detection rates of the other veins were lower: PMV, 65%; right ARV, 45%; left ARV, 15%; right PRV, 10%; left PRV, 30%; and SV, 0%. The average scores for AMV, PMV, right ARV, left ARV, right PRV, left PRV, and SV were 0.98, 0.24, 0.20, 0.08, 0.08, 0.14, and 0, respectively. CONCLUSION: SWI of the spine is feasible. The extrinsic spinal veins can be visualized by SWI without using contrast materials. PMID- 20027571 TI - Short echo time MR spectroscopy of brain tumors: grading of cerebral gliomas by correlation analysis of normalized spectral amplitudes. AB - PURPOSE: To process single voxel spectra of low- and high-grade gliomas. To propose correlation analysis of the scatter plots of normalized spectral amplitudes as a pattern recognition tool for the classification (grading) of brain tumors. To propose a spectrum processing approach that improves the differentiation of proton spectra with dominating macromolecule and lipid peaks. MATERIALS AND METHODS: LCModel was used to process spectra. Mean metabolite concentrations and mean normalized spectra were obtained for normal white matter and for gliomas. The mean spectra of macromolecules and lipids (ML) in the range 1.4-0.9 ppm, and mean difference spectra (DS) without ML and lactate were computed. Correlation analysis of the scatter plot of the patient and mean normalized spectral amplitudes and dispersion of the scatter plot points were used for classification and grading of tumors. RESULTS: It was found advantageous to perform the classifications using DS spectra. The shape of ML spectrum and concentration of tCr seem to be a good markers for glioma grade. CONCLUSION: Combining a qualitative comparison of the patient and mean DS spectra of the tumors using correlation analysis of normalized spectra amplitudes with a quantitative comparison of metabolite concentrations is a powerful tool in studying brain lesions. PMID- 20027572 TI - Objective and subjective evaluation of adaptive speech enhancement methods for functional MRI. AB - PURPOSE: To recover speech corrupted by functional magnetic resonance imaging (fMRI) acoustic noise using two-channel adaptive speech enhancement techniques. MATERIALS AND METHODS: Speech corrupted by noise generated from a 3 T MRI scanner was recorded using diffuse-field microphones and a data acquisition board. Multiband and subband adaptive speech enhancement methods are used to recover the speech signal from the recordings. Normalized least mean squares (NLMS) algorithm was used for updating the filter coefficients in each band. RESULTS: The methods are successful in enhancing the speech quality. They are successful in improving the convergence rate of the adaptive filter. Multiband and subband methods have a similar performance in terms of noise reduction and in the subjective tests. The subband method introduces less speech distortion compared to the multiband method. The subband method requires a lower number of computations per sample. CONCLUSION: Adaptive speech enhancement techniques are effective in reducing fMRI background noise in the recordings. Based on the analysis, we conclude that subband-based methods are more suited for enhancing speech corrupted by fMRI noise. PMID- 20027573 TI - Hybrid of opposite-contrast MRA of the brain by combining time-of-flight and black-blood sequences: initial experience in major trunk stenoocclusive diseases. AB - PURPOSE: To assess the feasibility of a new MR angiography (MRA) technique named hybrid of opposite-contrast MRA (HOP MRA) that combined the time-of-flight (TOF) MRA with a flow-sensitive black-blood (FSBB) sequence in the diagnosis of major trunk stenoocclusive diseases. MATERIALS AND METHODS: On a 1.5 Tesla imager using a dual-echo three-dimensional (3D)-gradient-echo sequence, we obtained the first echo for TOF MRA followed by the second echo for FSBB. We then subtracted the FSBB data set from that of TOF MRA followed by maximum intensity projection. In four normal volunteers and 19 patients with chronic stenoocclusive disease of the major trunk, we performed HOP MRA along with 3D-TOF MRA and compared the findings. RESULTS: In the volunteer group, the HOP MRA technique improved the demonstration of distal arterial branches. In 12 of the 19 patients, the HOP MRA better visualized branches distal to the lesion as well as distal branches of normal trunks than 3D-TOF MRA, while both techniques provided equivalent depiction of branches distal to the lesion but better depiction of normal distal branches in three patients. CONCLUSION: The HOP-MRA technique is promising in major trunk stenoocclusive diseases as it better demonstrates distal branches probably representing collaterals than 3D-TOF MRA. PMID- 20027574 TI - Neural specificity of acupuncture stimulation at pericardium 6: evidence from an FMRI study. AB - PURPOSE: To investigate the neural specificity of pericardium PC6, with the same meridian acupoint PC7 and a treatment-irrelevant acupoint GB37 as separate controls. MATERIALS AND METHODS: Functional magnetic resonance imaging (MRI) of the whole brain was performed in 36 healthy subjects receiving acupuncture at three acupoints, respectively: the study acupoint (PC6), and control acupoints (PC7 and GB37). A novel nonrepeated event-related (NRER) design paradigm was applied to separately detect neural activities related to different stages of acupuncture (needling manipulation and post-acupuncture rest epoch). Psychophysical responses (Deqi sensations) were also assessed. RESULTS: Neuroimaging studies of PC6 presented extensive signal attenuations in the cerebrocerebellar and subcortical areas, whereas acupuncture at GB37 induced widespread signal potentiations. In addition, acupuncture at PC6, in comparison with stimulations at PC7 and GB37, selectively evoked neural responses of the insula, hypothalamus, and flocculonodular lobe of cerebellum (nodulus and uvula). CONCLUSION: These findings may provide preliminary evidence for specific involvements of the cerebellar-hypothalamus and insula following acupuncture at PC6, which underlies the autonomic regulation of vestibular functions. The predominantly time-prolonged deactivations in these areas may also serve the clinical efficacy of PC6 in producing a sedative or tranquilizing effect in antiemetic treatment. PMID- 20027575 TI - Breathhold-regulated blood oxygenation level-dependent (BOLD) MRI of human brain at 3 tesla. AB - PURPOSE: To investigate the cerebrovascular response to repeated breathhold challenges using blood oxygenation level-dependent (BOLD) MRI at 3T and compare the results with previous data at 1.5T. MATERIALS AND METHODS: Six normal volunteers and six patients with brain tumors were recruited for this 3T study. For the normal group, BOLD MRI during repeated breathholds of different durations (five to 30 seconds) were acquired. Maximum signal change, full-width at half maximum (FWHM) and onset time (defined as the time to the first half-maximum) were determined by curve fitting. The fractional activation volume was also calculated. Patients performed a 10- or 15-second breathhold paradigm according to individual capability. RESULTS: Significant BOLD signal increases in the gray matter for a breathhold period as short as 5 seconds at 3T, instead of 10 seconds at 1.5T. The fractional activation volume vs. breathhold duration reached a plateau of 49.54 +/- 7.26% at 15 seconds at 3T, which was higher and shorter than that at 1.5T. The maximum signal changes were significantly larger (a 69% increase) at 3T than at 1.5T. In the patient group, there were BOLD signal increases in gray matter but not in tumor bulk or perifocal edema, which agreed with the results previously found at 1.5T. CONCLUSION: BOLD MRI at 3T is more sensitive for detecting breathhold-regulated signal changes than at 1.5T, which allows a shorter and more feasible breathhold paradigm for clinical applications in patients with brain tumors. PMID- 20027576 TI - Computer-aided detection of metastatic brain tumors using automated three dimensional template matching. AB - PURPOSE: To demonstrate the efficacy of an automated three-dimensional (3D) template matching-based algorithm in detecting brain metastases on conventional MR scans and the potential of our algorithm to be developed into a computer-aided detection tool that will allow radiologists to maintain a high level of detection sensitivity while reducing image reading time. MATERIALS AND METHODS: Spherical tumor appearance models were created to match the expected geometry of brain metastases while accounting for partial volume effects and offsets due to the cut of MRI sampling planes. A 3D normalized cross-correlation coefficient was calculated between the brain volume and spherical templates of varying radii using a fast frequency domain algorithm to identify likely positions of brain metastases. RESULTS: Algorithm parameters were optimized on training datasets, and then data were collected on 22 patient datasets containing 79 total brain metastases producing a sensitivity of 89.9% with a false positive rate of 0.22 per image slice when restricted to the brain mass. CONCLUSION: Study results demonstrate that the 3D template matching-based method can be an effective, fast, and accurate approach that could serve as a useful tool for assisting radiologists in providing earlier and more definitive diagnoses of metastases within the brain. PMID- 20027577 TI - Diffusion-weighted images differentiate benign from malignant thyroid nodules. AB - PURPOSE: To reveal the possible role of diffusion-weighted images (DWI) in the differential diagnosis of benign and malignant thyroid nodules by comparing the results of fine-needle aspiration cytology (FNAC). MATERIALS AND METHODS: In an 18-month period (December 2005 to May 2007), 27 cases with benign thyroid nodules with a total of 52 benign nodules, nine cases with thyroid gland malignancy, and 24 healthy control cases were included in the study. Cases that were indicated to undergo to FNAC examination and sent by a clinician for biopsy to the radiology unit were included in the study to assess the cytopathologic confirmation of the clinic, ultrasonographic, and magnetic resonance imaging (MRI) findings. RESULTS: The mean apparent diffusion coefficient (ADC) values of thyroid nodules were 2745.3 +/- 601.1 x 10(-6) mm(2)/s (1605-3899 x 10(-6)mm(2)/s) in the benign group and 695.2 +/- 312.5 x 10(-6)mm(2)/s (165-1330 x 10(-6)mm(2)/s) in the malignant group. Normal thyroid tissues had mean ADC values of 1344.1 +/- 276.4 x 10(-6) mm(2)/s (1015-1764 x 10(-6)mm(2)/s). The ADC values of three subgroups were significantly different (P = 0.0001). A reduced ADC was observed in most types of malignant tumors due to the consequent decrease of the extracellular extravascular space. CONCLUSION: Our preliminary results showed that ADC values of nodules may provide useful data about the nature of a thyroid nodule. PMID- 20027578 TI - Early identification of aortic valve sclerosis using iron oxide enhanced MRI. AB - PURPOSE: To test the ability of MION-47 enhanced MRI to identify tissue macrophage infiltration in a rabbit model of aortic valve sclerosis (AVS). MATERIALS AND METHODS: The aortic valves of control and cholesterol-fed New Zealand White rabbits were imaged in vivo pre- and 48 h post-intravenous administration of MION-47 using a 1.5 Tesla (T) MR clinical scanner and a CINE fSPGR sequence. MION-47 aortic valve cusps were imaged ex vivo on a 3.0T whole body MR system with a custom gradient insert coil and a three-dimensional (3D) FIESTA sequence and compared with aortic valve cusps from control and cholesterol fed contrast-free rabbits. Histopathological analysis was performed to determine the site of iron oxide uptake. RESULTS: MION-47 enhanced the visibility of both control and cholesterol-fed rabbit valves in in vivo images. Ex vivo image analysis confirmed the presence of significant signal voids in contrast administered aortic valves. Signal voids were not observed in contrast-free valve cusps. In MION-47 administered rabbits, histopathological analysis revealed iron staining not only in fibrosal macrophages of cholesterol-fed valves but also in myofibroblasts from control and cholesterol-fed valves. CONCLUSION: Although iron oxide labeling of macrophage infiltration in AVS has the potential to detect the disease process early, a macrophage-specific iron compound rather than passive targeting may be required. PMID- 20027579 TI - Measuring the heart in pulmonary arterial hypertension (PAH): implications for trial study size. AB - PURPOSE: To calculate the sample size for a theoretical pulmonary arterial hypertension (PAH) randomized controlled trial (RCT) by using cardiovascular magnetic resonance (CMR) imaging to determine the repeatability of measures between two scans. MATERIALS AND METHODS: Two same-day examinations from 10 PAH patients were analyzed manually and semiautomatically. Study size was calculated from the standard deviation (SD) of repeatability. Different approaches to right ventricle (RV) mass were investigated, agreement between methods tested and interobserver reproducibility measured by Bland-Altman analysis to explore how the PAH heart might be best measured. RESULTS: Repeatability was good for almost all manually-measured indices but poor for semiautomated measurement of RV mass and left-ventricle (LV) end-diastolic volume (EDV). Thus, for an RCT (power, 80%; significance level, 5%) analyzing "outcome" indices (RVEDV, LVEDV, RV ejection fraction, and RV mass; anticipated change: 10 mL, 10 mL, 3%, and 10 g, respectively) manually, 34 patients are required compared to 78 if analysis is semiautomated. RV mass was repeatable if the interventricular septum was divided between ventricles or if wholly apportioned to the LV. Limits of agreement between manual and semiautomated analyses were unsatisfactory for RV measures and interobserver reproducibility was worse for semiautomated than manual analysis. CONCLUSION: Manual is more robust than semiautomated analysis and at present should be favored in RCTs in PAH as it leads to lower sample size requirements. PMID- 20027580 TI - Comprehensive MR evaluation of renal disease: added clinical value of quantified renal perfusion values over single MR angiography. AB - PURPOSE: To evaluate the diagnostic accuracy of quantified renal perfusion parameters in identifying and differentiating renovascular from renal parenchymal disease. MATERIALS AND METHODS: In all, 27 patients underwent renal perfusion measurements on a 3.0 T magnetic resonance imaging (MRI) system. Imaging was performed with a saturation recovery TurboFLASH sequence (TR/TE 177/0.93 msec, flip angle 12 degrees , 5 slices/sec). All patients also underwent high resolution MR angiography (MRA) (TR/TE 3.1/1.09, flip angle 23 degrees , spatial resolution 0.9 x 0.8 x 0.9 mm(3)). MR perfusion measurements were analyzed with a two-compartment model, quantifying the plasma flow (F(P))-a characteristic renal first-pass perfusion parameter. A receiver-operator characteristic analysis was used to determine the optimal threshold value for distinguishing normal and abnormal plasma flow values. Utilizing this cutoff, sensitivity and specificity of solitary MR perfusion measurements, MRA, and a diagnostic strategy combining the two were evaluated. RESULTS: Quantified MR perfusion values yielded a sensitivity of 100% and a specificity of 85% utilizing the optimal plasma flow threshold value of 150 mL/100 mL/min, whereas single MRA achieved a sensitivity of 51.9% and a specificity of 90%. Combining both methods enabled improved detection of renovascular and renoparenchymal disease with a sensitivity of 96.3% and specificity of 90%. CONCLUSION: In distinction to MRA, quantified MR perfusion measurements allow for the detection of pure renal parenchymal disorders. The combination of MRA with these perfusion measurements suggests an algorithm by which parenchymal and renovascular diseases may be reliably distinguished and the hemodynamic significance of the latter reliably determined. PMID- 20027581 TI - Comparison of lymph node metastases assessment with the use of USPIO-enhanced MR imaging at 1.5 T versus 3.0 T in a rabbit model. AB - PURPOSE: To prospectively compare the diagnostic performances of 1.5 T and 3.0 T ultrasmall superparamagnetic iron oxide (USPIO)-enhanced magnetic resonance imaging (MRI) for the detection of lymph node (LN) metastases in a rabbit model. MATERIALS AND METHODS: Experiments were approved by the animal care committee. VX2 carcinomas were implanted into the thighs of 18 rabbits 3 or 4 weeks before MRI examinations. T2-weighted 1.5 T and 3.0 T MRI was performed 24 hours after USPIO (2.6 mg/kg iron) administration. Two radiologists calculated signal intensity (SI) ratios (ie, the ratios of postcontrast to precontrast signal intensity) of each LN and also evaluated for the presence of a metastasis in the iliac and retroperitoneal LNs at 1.5 T and 3.0 T MRI. Student's t-test, receiver operating characteristic (ROC) curve analysis, and a Z test were used for the statistical analysis. RESULTS: Metastases were confirmed in 45 of 80 LNs by histopathology. For benign LNs the mean SI ratio at 1.5 T was higher (0.538) than at 3.0 T (0.463) (P = 0.004). In qualitative analysis, 1.5 T and 3.0 T USPIO enhanced MRI showed similar Az values (0.951 vs. 0.939; P = 0.296). The specificity of 3.0 T USPIO-enhanced MRI was higher (91.4% [32 of 35]) than that of 1.5 T imaging (82.9% [29 of 35]) (P = 0.013); however, the sensitivities (88.9% [40 of 45]) were the same in the both modalities. CONCLUSION: 3.0 T USPIO enhanced MRI shows a higher specificity as compared to 1.5 T imaging without a significant difference of sensitivity in a rabbit VX2 model. PMID- 20027582 TI - Imaging cerebral microbleeds using susceptibility weighted imaging: one step toward detecting vascular dementia. AB - PURPOSE: To monitor changes in the number of cerebral microbleeds (CMBs) in a longitudinal study of healthy controls (HC) and mild-cognitively impaired (MCI) patients using susceptibility weighted imaging (SWI). MATERIALS AND METHODS: SWI was used to image 28 HC and 75 MCI patients annually at 1.5 Tesla over a 4-year period. Magnitude and phase data were used to visualize CMBs for the first and last scans of 103 subjects. RESULTS: Preliminary analysis revealed that none of the 28 HC had more than three CMBs. In the 75 MCI patients, five subjects had more than three CMBs in both first and last scans, while one subject had more than three bleeds only in the last scan. In five of these six MCI patients, the number of CMBs increased over time and all six went on to develop progressive cognitive impairment (PCI). Of the 130 total CMBs seen in the last scans of the six MCI cases, most were less than 4 mm in diameter. CONCLUSION: SWI can reveal small CMBs on the order of 1 mm in diameter and this technique can be used to follow their development longitudinally. Monitoring CMBs may be a means by which to evaluate patients for the presence of microvascular disease that leads to PCI. PMID- 20027583 TI - Direct comparison of sensitivity encoding (SENSE) accelerated and conventional 3D contrast enhanced magnetic resonance angiography (CE-MRA) of renal arteries: effect of increasing spatial resolution. AB - PURPOSE: To assess the effect of attaining higher spatial resolution in contrast enhanced magnetic resonance angiography (MRA) of renal arteries using parallel imaging, sensitivity encoding (SENSE), by comparing the SENSE contrast-enhanced (CE) MRA against a conventional CE-MRA protocol with identical scan times, injection protocol, and other acquisition parameters. MATERIALS AND METHODS: Numerical simulations and a direct comparison of SENSE-accelerated versus conventional acquisitions were performed. A total of 41 patients (18 male) were imaged using both protocols for a direct comparison. Both protocols used fluoroscopic triggering, centric encoding, breath-holding, equivalent injection protocol, and lasted approximately 30 seconds. RESULTS: Simulated point-spread functions were narrower for the SENSE protocol compared to the conventional protocol. In the patient study, although the SENSE protocol produced images with lower signal-to-noise ratio (SNR), image quality was better for all segments of the renal arteries. In addition, ringing of kidney parenchyma and renal artery blurring were significantly reduced in the SENSE protocol. Finally, reader confidence improved with the SENSE protocol. CONCLUSION: Despite a reduction in SNR, the higher-resolution SENSE CE-MRA provided improved image quality, reduced artifacts, and increased reader confidence compared to the conventional protocol. PMID- 20027584 TI - Scan-rescan reproducibility of carotid atherosclerotic plaque morphology and tissue composition measurements using multicontrast MRI at 3T. AB - PURPOSE: To evaluate interscan reproducibility of both vessel morphology and tissue composition measurements of carotid atherosclerosis using a fast, optimized, 3T multicontrast protocol. MATERIALS AND METHODS: A total of 20 patients with carotid stenosis >15% identified by duplex ultrasound were recruited for two independent 3T MRI (Philips) scans within one month. A multicontrast protocol including five MR sequences was applied: TOF, T1-/T2-/PD weighted and magnetization-prepared rapid acquisition gradient-echo (MP-RAGE). Carotid artery morphology (wall volume, lumen volume, total vessel volume, normalized wall index, and mean/maximum wall thickness) and plaque component size (lipid rich/necrotic core, calcification, and hemorrhage) were measured over two time points. RESULTS: After exclusion of images with poor image quality, 257 matched locations from 18 subjects were available for analysis. For the quantitative carotid morphology measurements, coefficient of variation (CV) ranged from 2% to 15% and intraclass correlation coefficient (ICC) ranged from 0.87 to 0.99. Except for maximum wall thickness (ICC = 0.87), all ICC were larger than 0.90. For the quantitative plaque composition measurements, the ICC of the volume and relative content of lipid rich/necrotic core and calcification were larger than 0.90 with CV ranging from 22% to 32%. CONCLUSION: The results from the multicontrast high-resolution 3T MR study show high reliability for carotid morphology and plaque component measurements. 3T MRI is a reliable tool for longitudinal clinical trials, with shorter scan time compared to 1.5T. PMID- 20027585 TI - Measuring aortic diameter with different MR techniques: comparison of three dimensional (3D) navigated steady-state free-precession (SSFP), 3D contrast enhanced magnetic resonance angiography (CE-MRA), 2D T2 black blood, and 2D cine SSFP. AB - PURPOSE: To compare nongated three-dimensional (3D) contrast-enhanced magnetic resonance angiography (CE-MRA) with 3D-navigated cardiac-gated steady-state free precession bright blood (3D-nav SSFP) and noncontrast 2D techniques for ascending aorta dimension measurements. MATERIALS AND METHODS: Twenty-five clinical exams were reviewed to evaluate the ascending aorta at 1.5T using: breathhold cine bright blood (SSFP), cardiac-triggered T2 black blood (T2 BB), axial 3D-nav SSFP, and nongated 3D CE-MRA. Three radiologists independently measured aortic size at three specified locations for each sequence. Means, SDs, interobserver correlation, and vessel edge sharpness were statistically evaluated. RESULTS: Measurements were greatest for 3D-nav SSFP and 3D CE-MRA and smallest for T2 BB. There was no significant difference between 3D-nav SSFP and 3D CE-MRA (P = 0.43 0.86), but significance was observed comparing T2 BB to all sequences. Interobserver agreement was uniformly >0.9, with T2 BB best, followed closely by 3D-nav SSFP and 2D cine SSFP, and 3D CE-MRA being the worst. Edge sharpness was significantly poorer for 3D CE-MRA compared to the other sequences (P < 0.001). CONCLUSION: If diameter measurements are the main clinical concern, 3D-nav SSFP appears to be the best choice, as it has a sharp edge profile, is easy to acquire and postprocess, and shows very good interobserver correlation. PMID- 20027586 TI - Susceptibility weighted imaging with multiple echoes. AB - PURPOSE: To extend susceptibility weighted imaging (SWI) to multiple echoes with an adapted homodyne filtering of phase images for the computation of venograms with improved signal to noise ratio (SNR) and contrast to noise ratio (CNR) and to produce high resolution maps of R(2) relaxation. MATERIALS AND METHODS: Three dimensional multi echo gradient echo data were acquired with five equidistant echoes ranging from 13 to 41 ms. The phase images of each echo were filtered with filter parameters adjusted to the echo time, converted into a phase mask, and combined with the corresponding magnitude images to obtain susceptibility weighted images. The individual images were then averaged. Conventional single echo data were acquired for comparison. Maps of R(2) relaxation rates were computed from the magnitude data. Field maps derived from the phase data were used to correct R(2) for the influences from background inhomogeneities of the static magnetic field. RESULTS: Compared with the single echo images, the combined images had an increase in SNR by 46% and an improvement in CNR by 34 to 80%, improved visibility of small venous vessels and reduced blurring along the readout direction. The R(2) values of different tissue types are in good agreement with values from the literature. CONCLUSION: Acquisition of SWI with multiple echoes leads to an increase in SNR and CNR and it allows the computation of high resolution maps of R(2) relaxation. PMID- 20027587 TI - Accuracy of phase contrast, black-blood, and bright-blood pulse sequences for measuring compliance and distensibility coefficients in a human-tissue mimicking phantom. AB - PURPOSE: To assess the accuracy of MR-derived luminal diameter variations and its implications for compliance (CC) and distensibility coefficients (DC) by comparison with high-resolution digital photography (HRDP) in a tissue-mimicking phantom with pulsatile flow. MATERIALS AND METHODS: Diameters, CC, and DC extracted using cine phase-contrast (CPC), cine bright-blood (CBrB), and a cine black-blood (CBB) sequence were compared. The diameter in the left-right direction was compared against HRDP, as the gold-standard. The experiments were performed using 256(2) and 512(2) matrix sizes. Bland-Altman analysis was performed to compare each sequence with the gold-standard in terms of diameter changes over the simulated cardiac cycle. RESULTS: The bias and 95% limits of agreement (LOA) for CBB and CBrB were comparable. The bias for CPC was larger, however, the LOA were comparable. Increasing spatial resolution improved agreement with HRDP for all sequences. CBrB-derived CC and DC were within 3% of the high resolution CBB values while CPC CC and DC were underestimated but still within 11%. CONCLUSION: CPC images were found to underestimate the luminal area over the cardiac cycle. CBrB-derived diameters were more accurate in diastole while CBB-derived diameters gave the best results in systole. CC and DC varied depending on the pulse sequence. PMID- 20027588 TI - Adaptive non-local means denoising of MR images with spatially varying noise levels. AB - PURPOSE: To adapt the so-called nonlocal means filter to deal with magnetic resonance (MR) images with spatially varying noise levels (for both Gaussian and Rician distributed noise). MATERIALS AND METHODS: Most filtering techniques assume an equal noise distribution across the image. When this assumption is not met, the resulting filtering becomes suboptimal. This is the case of MR images with spatially varying noise levels, such as those obtained by parallel imaging (sensitivity-encoded), intensity inhomogeneity-corrected images, or surface coil based acquisitions. We propose a new method where information regarding the local image noise level is used to adjust the amount of denoising strength of the filter. Such information is automatically obtained from the images using a new local noise estimation method. RESULTS: The proposed method was validated and compared with the standard nonlocal means filter on simulated and real MRI data showing an improved performance in all cases. CONCLUSION: The new noise-adaptive method was demonstrated to outperform the standard filter when spatially varying noise is present in the images. PMID- 20027589 TI - Diurnal stability of gamma-aminobutyric acid concentration in visual and sensorimotor cortex. AB - PURPOSE: To establish the diurnal stability of edited magnetic resonance spectroscopy measurements of gamma-aminobutyric acid (GABA) in visual and sensorimotor regions of the brain. MATERIALS AND METHODS: GABA measurements were made in two regions of the brain (an occipital, "visual" region and a "sensorimotor" region centered on the precentral gyrus) using the MEGA-PRESS editing method, scanning eight healthy adults at five timepoints during a single day. GABA concentration was quantified from the ratio of the GABA integral to the unsuppressed water signal. RESULTS: No significant effect of time on GABA concentration was seen (P = 0.35). GABA was shown to be significantly more concentrated in visual regions than in sensorimotor regions (1.10 i.u. and 1.03 i.u., respectively; P = 0.050). Coefficients of variability (CVs) across all subjects of 9.1% and 12% (visual and sensorimotor) were significantly higher than mean within-subjects CVs of 6.5% and 8.8. CONCLUSION: This study demonstrates the excellent reproducibility of MEGA-PRESS detection of GABA, demonstrating that the method is sufficiently sensitive to detect inter-subject variability, and suggests that (within the sensitivity limits of current measurements) time of day can be ignored in the design of MRS studies of visual and sensorimotor regions. PMID- 20027590 TI - Three-dimensional (3D) visualization of endolymphatic hydrops after intratympanic injection of Gd-DTPA: optimization of a 3D-real inversion-recovery turbo spin echo (TSE) sequence and application of a 32-channel head coil at 3T. AB - PURPOSE: To enable volume visualization of endolymphatic hydrops of Meniere's disease via a volume rendering (VR) technique, a three-dimensional (3D) inversion recovery (IR) sequence with real reconstruction (3D-real IR) sequence after intratympanic injection of Gd-DTPA was optimized for higher spatial resolution using a 32-channel head coil at 3T. MATERIALS AND METHODS: Pulse sequence parameters were optimized using a diluted Gd-DTPA phantom. Then, 11 patients who had been clinically diagnosed with Meniere's disease and a patient with sudden hearing loss were scanned. Images were processed using commercially available 3D VR software. 3D-real IR data was processed to produce endolymph and perilymph fluid volume images in different colors. 3D-CISS data was processed to generate total fluid volume images. RESULTS: While maintaining a comparable signal-to noise ratio (SNR) and scan time, the voxel volume could be reduced from 0.4 x 0.4 x 2 mm(3) with a 12-channel coil to 0.4 x 0.4 x 0.8 mm(3) with a 32-channel coil. A newly-optimized protocol allowed the smooth, three-dimensional visualization of endolymphatic hydrops in all patients with Meniere's disease. CONCLUSION: Volumetrically separate visualization of endo-/perilymphatic space is now feasible in patients with Meniere's disease using an optimized 3D-real IR sequence, a 32-channel head coil, at 3T, after intratympanic administration of Gd DTPA. This will aid the understanding of the pathophysiology of Meniere's disease. PMID- 20027591 TI - A population-specific symmetric phase model to automatically analyze susceptibility-weighted imaging (SWI) phase shifts and phase symmetry in the human brain. AB - PURPOSE: To create a population-specific symmetric phase model and to evaluate the susceptibility-weighted imaging (SWI) phase in terms of phase shift using different segmentation methods (manual and automatic) and phase shift symmetry, which is expected as a marker for lateralized Parkinson's disease (PD) symptoms. MATERIALS AND METHODS: SWI and T(1)-weighted data from 25 PD patients and five healthy controls were acquired on a 3T MRI system. A population-specific, symmetric phase model was developed. Regions of interest (ROIs) were defined manually on the phase model, manually on each individual data set, and automatically using model-based segmentation (MBS). Manually- and MBS-defined ROIs were compared using kappa values, and left-right phase symmetry was evaluated using correlation analysis. RESULTS: Independent of the analysis method, a phase increase from the anterior to the posterior putamen, and the average phase value relationship substantia nigra > globus pallidus > red nucleus was found. Phase symmetry analysis shows a difference between lateralized and symmetric PD. CONCLUSION: The symmetric phase model helps to analyze phase data with similar accuracy, but a greatly reduced tracing effort compared to individual tracing and also allows evaluating left-right phase symmetries. PMID- 20027592 TI - Cerebral asymmetry in patients with schizophrenia: a voxel-based morphometry (VBM) and diffusion tensor imaging (DTI) study. AB - PURPOSE: To evaluate the differences in gray- and white-matter asymmetry between schizophrenia patients and normal subjects. MATERIALS AND METHODS: Forty-eight right-handed patients with chronic schizophrenia (24 males and 24 females) and 48 right-handed age- and sex-matched healthy controls (24 males and 24 females) were included in this study. The effects of diagnosis on gray-matter volume asymmetry and white-matter fractional anisotropy (FA) asymmetry were evaluated with use of voxel-based morphometry (VBM) and voxel-based analysis of FA maps derived from diffusion tensor imaging (DTI), respectively. RESULTS: The mean gray- and white matter volumes were significantly smaller in the schizophrenia group than in the control group. The voxel-based morphometry (VBM) showed no significant effect of diagnosis on gray-matter volume asymmetry. The voxel-based analysis of DTI also showed no significant effect of diagnosis on white-matter FA asymmetry. CONCLUSION: Our results of voxel-based analyses showed no significant differences in either gray-matter volume asymmetry or white-matter FA asymmetry between schizophrenia patients and normal subjects. PMID- 20027593 TI - Improved correction of spatial inhomogeneities of surface coils in quantitative analysis of first-pass myocardial perfusion imaging. AB - PURPOSE: To test whether image normalization using either a separate 3D proton density (PD)-weighted prescan, or 2D PD-weighted images prior to the perfusion series, improves correction of differences in spatial sensitivity induced by radiofrequency (RF) surface receiver coils. Originally, this correction was applied using the baseline signal in the myocardium before arrival of the contrast agent. This is of importance, as quantitative analysis of magnetic resonance (MR) myocardial perfusion using deconvolution with the arterial input assumes equal signal sensitivity over the heart. MATERIALS AND METHODS: First pass myocardial perfusion measurements were obtained in 13 patients without known coronary artery disease. Absolute perfusion values were assessed for 18 myocardial segments without any normalization and using the three different normalization methods. RESULTS: Using 2D or 3D PD-weighted normalization, similar mean perfusion values were found, but with reduced spatial variance over the 18 segments. The relative dispersion of perfusion at rest was 23% and 35% for the 3D prescan normalization and the baseline normalization, respectively. With 2D and 3D PD-weighted prescan normalization the relative dispersion was closer to the expected physiological heterogeneity. CONCLUSION: PD-weighted prescan normalization proved to be a valuable addition to quantitative analysis of myocardial perfusion, and better than baseline-based normalization. PMID- 20027594 TI - B1 transmission-field inhomogeneity and enhancement ratio errors in dynamic contrast-enhanced MRI (DCE-MRI) of the breast at 3T. AB - PURPOSE: To quantify B(1) transmission-field inhomogeneity in breast imaging of normal volunteers at 3T using 3D T(1)-weighted spoiled gradient echo and to assess the resulting errors in enhancement ratio (ER) measured in dynamic contrast-enhanced MRI (DCE-MRI) studies of the breast. MATERIALS AND METHODS: A total of 25 volunteers underwent breast imaging at 3T and the B(1) transmission fields were mapped. Gel phantoms that simulate pre- and postcontrast breast tissue T(1) were developed. The effects of B(1)-field inhomogeneity on ER, as measured using a 3D spoiled gradient echo sequence, were investigated by computer simulation and experiments on gel phantoms. RESULTS: It was observed that by using the patient orientation and MR scanner employed in this study, the B(1) transmission-field field is always reduced toward the volunteer's right side. The median B(1)-field in the right breast is reduced around 40% of the expected B(1) field. For some volunteers the amplitude was reduced by more than 50%. Computer simulation and experiment showed that a reduction in B(1)-field decreases ER. This reduction increases with both B(1)-field error and contrast agent uptake. CONCLUSION: B(1) transmission-field inhomogeneity is a critical issue in breast imaging at 3T and causes errors in quantifying ER. These errors would be sufficient to reduce the conspicuity of a malignant lesion and could result in reduced sensitivity for cancer detection. PMID- 20027595 TI - Flow-independent T(2)-prepared inversion recovery black-blood MR imaging. AB - PURPOSE: To develop a magnetization preparation method to achieve robust, flow independent blood suppression for cardiac and vascular magnetic resonance imaging (MRI). MATERIALS AND METHODS: T(2)Prep-IR sequence consists of a T(2) preparation followed by a nonselective adiabatic inversion pulse. T(2)Prep separates the initial longitudinal magnetization of arterial wall from lumen blood. After the inversion recovery pulse the imaging acquisition is then delayed for a period that allows the blood signal to approach the zero-crossing point. Compared to the conventional double inversion recovery (DIR) preparation, T(2)Prep-IR prepares all the spins regardless of their velocity and direction. T(2)Prep-IR was incorporated into the fast spin echo and fast gradient echo acquisition sequences and images in various planes were acquired in the carotid arteries, thoracic aorta, and heart of normal volunteers. Blood suppression and image quality were compared qualitatively between two different preparations. RESULTS: For in-plane flow carotid images, persistent flow-related artifacts on the DIR images were removed with T(2)Prep-IR. For cardiac applications, T(2)Prep-IR provided robust blood suppression regardless of the flow direction and velocity, including the cardiac long-axis views and the aorta that are often problematic with DIR. CONCLUSION: T(2)Prep-IR may overcome the flow dependence of DIR by providing robust flow-independent black-blood images. PMID- 20027596 TI - Dual-echo arteriovenography imaging with 7T MRI. AB - PURPOSE: To implement a dual-echo sequence MRI technique at 7T for simultaneous acquisition of time-of-flight (TOF) MR angiogram (MRA) and blood oxygenation level-dependent (BOLD) MR venogram (MRV) in a single MR acquisition and to compare the image qualities with those acquired at 3T. MATERIALS AND METHODS: We implemented a dual-echo sequence with an echo-specific k-space reordering scheme to uncouple the scan parameter requirements for MRA and MRV at 7T. The MRA and MRV vascular contrast was enhanced by maximally separating the k-space center regions acquired for the MRA and MRV and by adjusting and applying scan parameters compatible between the MRA and MRV. The same imaging sequence was implemented at 3T. Four normal subjects were imaged at both 3T and 7T. MRA and MRV at 7T were reconstructed both with and without phase-mask filtering and were compared quantitatively and qualitatively with those at 3T with phase-mask filtering. RESULTS: The depiction of small cortical arteries and veins on MRA and MRV at 7T was substantially better than that at 3T, due to about twice higher contrast-to-noise ratio (CNR) for both arteries (164 +/-57 vs. 77 +/- 26) and veins (72 +/- 8 vs. 36 +/- 6). Even without use of the phase-masking filtering, the venous contrast at 7T (65 +/- 7) was higher than that with the filtering at 3T (36 +/- 6). CONCLUSION: The dual-echo arteriovenography technique we implemented at 7T allows the improved visualization of small vessels in both the MRA and MRV because of the greatly increased signal-to-noise ratio (SNR) and susceptibility contrast, compared to 3T. PMID- 20027598 TI - Reconstruction from free-breathing cardiac MRI data using reproducing kernel Hilbert spaces. AB - This paper describes a rigorous framework for reconstructing MR images of the heart, acquired continuously over the cardiac and respiratory cycle. The framework generalizes existing techniques, commonly referred to as retrospective gating, and is based on the properties of reproducing kernel Hilbert spaces. The reconstruction problem is formulated as a moment problem in a multidimensional reproducing kernel Hilbert spaces (a two-dimensional space for cardiac and respiratory resolved imaging). Several reproducing kernel Hilbert spaces were tested and compared, including those corresponding to commonly used interpolation techniques (sinc-based and splines kernels) and a more specific kernel allowed by the framework (based on a first-order Sobolev RKHS). The Sobolev reproducing kernel Hilbert spaces was shown to allow improved reconstructions in both simulated and real data from healthy volunteers, acquired in free breathing. PMID- 20027599 TI - Quantitative T2 analysis: the effects of noise, regularization, and multivoxel approaches. AB - Typical quantitative T2 (qT2) analysis involves creating T2 distributions using a regularized algorithm from region-of-interest averaged decay data. This study uses qT2 analysis of simulated and experimental decay signals to determine how (a) noise-type, (b) regularization, and (c) region-of-interest versus multivoxel analyses affect T2 distributions. Our simulations indicate that regularization causes myelin water fraction and intra/extracellular water geometric mean T2 underestimation that worsens as the signal-to-noise ratio decreases. The underestimation was greater for intra/extracellular water geometric mean T2 measures using Rician noise. Simulations showed significant differences between myelin water fractions determined using region-of-interest and multivoxel approaches compared to the true value. The nonregularized voxel-based approach gave the most accurate measure of myelin water fraction and intra/extracellular water geometric mean T2 for a given signal-to-noise ratio and noise type. Additionally, multivoxel analysis provides important information about the variability of the analysis. Results obtained from in vivo rat data were similar to our simulation results. In each case, a nonregularized, multivoxel analysis provided myelin water fractions significantly different from the regularized approaches and obtained the largest myelin water fraction. We conclude that quantitative T2 analysis is best performed using a nonregularized, multivoxel approach. PMID- 20027600 TI - Comment on ADC reductions in postcontrast breast tumors. PMID- 20027601 TI - Screening for depression and anxiety: correlates of non-response and cohort attrition in the Netherlands study of depression and anxiety (NESDA). AB - A major problem in the analysis of attrition of cohorts in studies on mental health problems is that data on those who do not participate at the outset of a study are largely unavailable. It is not known how underlying psychopathology affects the first stages of screening where non-response and selectivity are usually highest. This article presents results of one of the centres of the Netherlands Study of Depression and Anxiety (NESDA), a longitudinal study aimed at describing the long-term course and consequences of depression and anxiety disorders. The aim is to describe the different ways of attrition during the first NESDA-wave in a cohort of patients aged 18-65 years of the Registration Network Groningen and to analyse whether attrition is related to gender, age and psychopathology as recorded in general practice. The attrition of the study cohort (n = 8475) was highest during the first stages, eventually leading to a population of 169 patients only who participated in the full NESDA-programme. Probabilities of transition from one stage of the screening process to the next were regressed on selected background variables using binary logistic regression. Correlates of participation were being female and being older (>40). Psychopathology was an important variable in the formation of the initial sample cohort, but only had a weak influence on patient response to the screening questionnaire. Study design factors had a stronger impact on the changing composition of the cohort at each screening stage compared to patient factors. PMID- 20027602 TI - Post-traumatic growth: finding positive meaning in cancer survivorship moderates the impact of intrusive thoughts on adjustment in younger adults. AB - OBJECTIVE: We examined whether post-traumatic growth would moderate the impact of intrusive thoughts on a range of dimensions of well-being in a sample of younger adult survivors of various types of cancer. METHODS: 167 participants completed questionnaires regarding intrusive thoughts, post-traumatic growth, mental and physical health-related quality of life, positive and negative affect, life satisfaction, and spiritual well-being. Multiple regression analyses controlling for relevant background and cancer-related variables tested the interaction effects of post-traumatic growth and intrusive thoughts. RESULTS: Intrusive thoughts were related to poorer adjustment on all indices except physical health related quality of life. However, post-traumatic growth moderated the effects of intrusive thoughts on positive and negative affect, life satisfaction, and spiritual well-being in a protective fashion. That is, for those higher in post traumatic growth, higher levels of intrusive thoughts were related to better adjustment. CONCLUSIONS: The positive meaning that individuals assign to their cancer experience as reflected in their reports of post-traumatic growth appears to be important in determining the impact of intrusive thoughts on post-cancer adjustment. PMID- 20027603 TI - Enhanced expression of proproliferative and antiapoptotic genes in ulcerative colitis-associated neoplasia. AB - BACKGROUND: Inflammatory bowel diseases including long-standing ulcerative colitis (UC) have an increased risk of evolving into colorectal cancer (CRC). The overexpression of some proproliferative and antiapoptotic genes, such as survivin, telomerase catalytic subunit (hTERT), integrin-linked kinase (ILK), and regulatory factors c-MYB and Tcf-4, has been implicated in the development and progression of several human malignancies including CRC. METHODS: In this study we analyzed the expression alterations of these markers and proinflammatory enzymes cyclooxygenase 2 (COX-2) and inducible nitric oxide synthase (iNOS) during the transition of colonic mucosa from chronic inflammation to epithelial neoplasia in biopsies of UC patients using quantitative real-time polymerase chain reaction and immunohistochemistry; additionally, we compared the expression profiles of this gene panel in samples of patients with CRC after tumor resection and in human tumor xenografts of SW620 malignant colonic cells. RESULTS: The transcript levels of survivin, c-MYB, COX-2, iNOS, and Tcf-4 showed a statistically significant increase during neoplastic transformation of UC patient colonic mucosa, whereas hTERT and ILK were not elevated. In contrast, the specimens of CRC showed upregulated expression of not only survivin, c-MYB, Tcf 4, COX-2, and iNOS but also hTERT. A similar expression profile was observed in human tumor xenografts in which all transcripts with the exception of c-MYB were upregulated. CONCLUSIONS: These results suggest that telomerase and ILK activation occurs during the later stages of carcinoma progression, whereas upregulation of survivin, c-MYB, and Tcf-4 is a feature of the early stage of development of neoplasia, and thus, they might serve as early indicators for UC associated colorectal carcinogenesis. PMID- 20027604 TI - Preserved Na(+)/H(+) exchanger isoform 3 expression and localization, but decreased NHE3 function indicate regulatory sodium transport defect in ulcerative colitis. AB - BACKGROUND: A major causative factor of diarrhea in ulcerative colitis (UC) patients is the loss of Na(+) absorptive capacity of the inflamed colonic mucosa. Potential contributing mechanisms include reduced driving force for active transport, and impaired expression, mislocalization, or defective transport function of Na(+) absorptive proteins. We therefore studied the expression, brush border membrane (BBM) localization, and transport capacity of the major intestinal Na(+) absorptive protein, the Na(+)/H(+) exchanger isoform 3 (NHE3) in biopsies from UC patients. METHODS: In UC and control biopsies, inflammation was graded histologically, NHE3, tumor necrosis factor alpha (TNF-alpha), villin, as well as other housekeeping genes were analyzed by quantitative real-time polymerase chain reaction (PCR), BBM localization of NHE3 determined by immunohistochemistry, and confocal microscopy. Na(+) absorptive capacity was assessed by (22)Na(+) isotope fluxes and NHE3 transport activity measured microfluorometrically in BCECF-loaded surface colonocytes within isolated crypts. RESULTS: In mildly, moderately, and severely inflamed sigmoid colon of UC patients, neither NHE3 mRNA expression nor the abundance of NHE3 in the BBM was significantly altered compared to other structural components of the BBM. However, Na(+) absorption was strongly reduced by approximately 80% and acid activated NHE3 transport activity was significantly decreased in the surface cells of sigmoid colonic crypts even in moderately inflamed mucosa. CONCLUSIONS: In the colonic mucosa of patients with active UC, NHE3 transport capacity was found significantly decreased despite correct NHE3 location and abundance in the brush border, independent of current treatment. These findings suggest functional NHE3 transport as a novel factor for inflammatory diarrhea in UC patients. PMID- 20027605 TI - Periportal cuffing in inflammatory bowel diseases: mystery of stars and stripes. PMID- 20027606 TI - Sequential steps underlying neuronal plasticity induced by a transient exposure to gabazine. AB - Periods of intense electrical activity can initiate neuronal plasticity leading to long lasting changes of network properties. By combining multielectrode extracellular recordings with DNA microarrays, we have investigated in rat hippocampal cultures the temporal sequence of events of neuronal plasticity triggered by a transient exposure to the GABA(A) receptor antagonist gabazine (GabT). GabT induced a synchronous bursting pattern of activity. The analysis of electrical activity identified three main phases during neuronal plasticity induced by GabT: (i) immediately after termination of GabT, an early synchronization (E-Sync) of the spontaneous electrical activity appears that progressively decay after 3-6 h. E-Sync is abolished by inhibitors of the ERK1/2 pathway but not by inhibitors of gene transcription; (ii) the evoked response (induced by a single pulse of extracellular electrical stimulation) was maximally potentiated 3-10 h after GabT (M-LTP); and (iii) at 24 h the spontaneous electrical activity became more synchronous (L-Sync). The genome-wide analysis identified three clusters of genes: (i) an early rise of transcription factors (Cluster 1), primarily composed by members of the EGR and Nr4a families, maximally up-regulated 1.5 h after GabT; (ii) a successive up-regulation of some hundred genes, many of which known to be involved in LTP (Cluster 2), 3 h after GabT likely underlying M-LTP. Moreover, in Cluster 2 several genes coding for K(+) channels are down-regulated at 24 h. (iii) Genes in Cluster 3 are up regulated at 24 h and are involved in cellular homeostasis. This approach allows relating different steps of neuronal plasticity to specific transcriptional profiles. PMID- 20027607 TI - Brief communication: Reaction to fire by savanna chimpanzees (Pan troglodytes verus) at Fongoli, Senegal: Conceptualization of "fire behavior" and the case for a chimpanzee model. AB - The use and control of fire are uniquely human traits thought to have come about fairly late in the evolution of our lineage, and they are hypothesized to correlate with an increase in intellectual complexity. Given the relatively sophisticated cognitive abilities yet small brain size of living apes compared to humans and even early hominins, observations of wild chimpanzees' reactions to naturally occurring fire can help inform hypotheses about the likely responses of early hominins to fire. We use data on the behavior of savanna chimpanzees (Pan troglodytes verus) at Fongoli, Senegal during two encounters with wildfires to illuminate the similarities between great apes and humans regarding their reaction to fire. Chimpanzees' close relatedness to our lineage makes them phylogenetically relevant to the study of hominid evolution, and the open, hot and dry environment at Fongoli, similar to the savanna mosaic thought to characterize much of hominid evolution, makes these apes ecologically important as a living primate model as well. Chimpanzees at Fongoli calmly monitor wildfires and change their behavior in anticipation of the fire's movement. The ability to conceptualize the "behavior" of fire may be a synapomorphic trait characterizing the human-chimpanzee clade. If the cognitive underpinnings of fire conceptualization are a primitive hominid trait, hypotheses concerning the origins of the control and use of fire may need revision. We argue that our findings exemplify the importance of using living chimpanzees as models for better understanding human evolution despite recently published suggestions to the contrary. PMID- 20027608 TI - Brief communication: Dynamic plantar pressure distribution during locomotion in Japanese macaques (Macaca fuscata). AB - To better place the form and motion of the human foot in an evolutionary context, understanding how foot motions change when quadrupeds walk bipedally can be informative. For this purpose, we compared the pressures beneath the foot during bipedal and quadrupedal walking in Japanese macaques (Macaca fuscata). The pressure at nine plantar regions was recorded using a pressure mat (120 Hz), while the animals walked on a level walkway at their preferred speeds. The results revealed substantial differences in foot use between the two modes of locomotion, and some features observed during bipedal walking resembled human gait, such as the medial transfer of the center of pressure (COP), abrupt declines in forefoot pressures, and the increased pressure beneath the hallux, all occurring during the late-stance phase. In particular, the medial transfer of the COP, which is also observed in bonobos (Vereecke et al.: Am J Phys Anthropol 120 (2003) 373-383), was due to a biomechanical requirement for a hind limb dominant gait, such as bipedal walking. Features shared by bipedal and quadrupedal locomotion that were quite different from human locomotion were also observed: the heel never contacted the ground, a foot longitudinal arch was absent, the hallux was widely abducted, and the functional axis was on the third digit, not the second. PMID- 20027610 TI - Life and death in a civitas capital: metabolic disease and trauma in the children from late Roman Dorchester, Dorset. AB - The impact that "Romanization" and the development of urban centers had on the health of the Romano-British population is little understood. A re-examination of the skeletal remains of 364 nonadults from the civitas capital at Roman Dorchester (Durnovaria) in Dorset was carried out to measure the health of the children living in this small urban area. The cemetery population was divided into two groups; the first buried their dead organized within an east-west alignment with possible Christian-style graves, and the second with more varied "pagan" graves, aligned north-south. A higher prevalence of malnutrition and trauma was evident in the children from Dorchester than in any other published Romano-British group, with levels similar to those seen in postmedieval industrial communities. Cribra orbitalia was present in 38.5% of the children, with rickets and/or scurvy at 11.2%. Twelve children displayed fractures of the ribs, with 50% of cases associated with rickets and/or scurvy, suggesting that rib fractures should be considered during the diagnosis of these conditions. The high prevalence of anemia, rickets, and scurvy in the Poundbury children, and especially the infants, indicates that this community may have adopted child rearing practices that involved fasting the newborn, a poor quality weaning diet, and swaddling, leading to general malnutrition and inadequate exposure to sunlight. The Pagan group showed no evidence of scurvy or rib fractures, indicating difference in religious and child-rearing practices but that both burial groups were equally susceptible to rickets and anemia suggests a shared poor standard of living in this urban environment. PMID- 20027611 TI - Brief communication: mitochondrial haplotype C4c confirmed as a founding genome in the Americas. AB - Mitochondrial DNA analysis of 31 unrelated Shuswap speakers from a previously poorly sampled region of North America revealed two individuals with haplogroups rarely found in the Americas, C4c and C1d. Comparison of the complete genomes of the two individuals with others found in the literature confirms that C4c is a founding haplotype and gives insight into the evolution of the C1d haplotype. This study demonstrates the importance of collecting and analyzing data from Native North Americans when addressing hypotheses about the peopling of the Americas. PMID- 20027616 TI - Construction of a dual affinity tagged allele of the Rb1 tumor suppressor gene in the mouse. AB - Loss of Rb1 tumor suppressor gene function is involved in the genesis of most human cancers. Novel therapies targeting Rb1 have been slow to develop because of our incomplete understanding of its molecular mechanisms of action. Rb1 protein (pRb) binds a host of cellular genes and proteins, and these molecular interactions mediate its various functions. Given the potential complexity of these molecular interactions and the lack of established methods for pRb purification, it has been difficult to systematically identify gene and protein interactions relevant to tumor suppression in different tissues in vivo. To address this limitation, we have generated a dual affinity tagged Rb1 allele in the mouse. The tagged allele functions as wild type and the encoded protein can be purified by tandem affinity chromatography. This allele will facilitate identification and characterization of native pRb molecular interactions in any tissue accessible in the mouse. PMID- 20027617 TI - Heat shock protein 2 promoter drives Cre expression in spermatocytes of transgenic mice. AB - We generated transgenic mouse line C57BL/6-Tg(Hspa2-cre)1Eddy/J (Hspa2-cre), which expresses cre-recombinase under the control of a 907-bp fragment of the heat shock protein 2 (Hspa2) gene promoter. Transgene expression was determined using Gt(ROSA)26Sor(tm1Sor)/J (ROSA26) and Tg(CAG-Bgeo/GFP)21Lbe/J (Z/EG) reporter strains and RT-PCR and immunohistochemistry assays. Hspa2-cre expression mimicked the spermatogenic cell-specific expression of endogenous HSPA2 within the testis, being first observed in leptotene/zygotene spermatocytes. Expression of the transgene also was detected at restricted sites in the brain, as occurs for endogenous HSPA2. Although the results of mating the Hspa2-cre mice to mice with a floxed Cdc2a allele indicated that some expression of the transgene occurs during embryogenesis, the Hspa2-cre mice provide a valuable new tool for assessing the roles of genes during and after meiotic prophase in pachytene spermatocytes. PMID- 20027618 TI - Lens morphogenesis is dependent on Pax6-mediated inhibition of the canonical Wnt/beta-catenin signaling in the lens surface ectoderm. AB - Lens formation in mouse is critically dependent on proper development of the retinal neuroectoderm that is located close beneath the head surface ectoderm. Signaling from the prospective retina triggers lens-specific gene expression in the surface-ectoderm. Supression of canonical Wnt/beta-catenin signaling in the surface ectoderm is one of the prerequisites for lens development because, as we show here, ectopic Wnt activation in the retina and lens abrogates lens formation. Wnt inhibiton is mediated by signals coming from the retina but its exact mechanism is unknown. We show that Pax6 directly controls expression of several Wnt inhibitors such as Sfrp1, Sfrp2, and Dkk1 in the presumptive lens. In accordance, absence of Pax6 function leads to aberrant canonical Wnt activity in the presumptive lens that subsequently impairs lens development. Thus Pax6 is required for down-regulation of canonical Wnt signaling in the presumptive lens ectoderm. PMID- 20027619 TI - Synchronous ipsilateral Warthin tumor encased by a separate mucoepidermoid carcinoma of the parotid gland: a case report and review of the literature. AB - Fine-needle aspiration (FNA) is a cost effective and low morbidity procedure in the initial assessment of salivary gland tumors. However, cytological assessment of ipsilateral synchronous tumors (which make up less than 0.3% of all salivary gland neoplasms) may pose diagnostic challenges. Therefore, a wholesome approach, including FNA with clinical and radiological correlation, is of utmost importance. Here, we report a unique case of Warthin tumor encased by a separate high-grade mucoepidermoid carcinoma that was first diagnosed on FNA. Another striking feature seen was the presence of chronic sialadenitis in the surrounding nonneoplastic salivary gland. The presence of two different neoplasms in the background of chronic sialadenitis raises the question of a possible causal relationship. Traditionally, there has been diagnostic difficulty when dealing with synchronous tumors of the salivary gland and the background of chronic sialadenitis may further complicate the diagnosis. FNA is very helpful and can give important cues to the diagnosis. PMID- 20027620 TI - Cyclic voltammetry: a new strategy for the evaluation of oxidative damage to bovine insulin. AB - Research on protein oxidative damage may give insight into the nature of protein functions and pathological conditions. In this work, the oxidative damage of bovine insulin on Au electrode was investigated by cyclic voltammetry (CV). The experimental results show that there are two anodic peaks for the oxidative damage of bovine insulin, which arise from the oxidation of the exposed disulfide bond S--S(CYS7A,CYS7B), forming sulfenic acid RSOH (1.20 V, vs. SCE), sulfinic acid RSO(2)H and sulfonic acid RSO(3)H (1.35 V, vs. SCE). These in vitro findings not only demonstrate the applicability of CV in simulating/evaluating the oxidative damage of nonredox proteins but also find two promising candidates (two anodic peaks) for measuring insulin. PMID- 20027621 TI - Amyloidogenic sequences in native protein structures. AB - Numerous short peptides have been shown to form beta-sheet amyloid aggregates in vitro. Proteins that contain such sequences are likely to be problematic for a cell, due to their potential to aggregate into toxic structures. We investigated the structures of 30 proteins containing 45 sequences known to form amyloid, to see how the proteins cope with the presence of these potentially toxic sequences, studying secondary structure, hydrogen-bonding, solvent accessible surface area and hydrophobicity. We identified two mechanisms by which proteins avoid aggregation: Firstly, amyloidogenic sequences are often found within helices, despite their inherent preference to form beta structure. Helices may offer a selective advantage, since in order to form amyloid the sequence will presumably have to first unfold and then refold into a beta structure. Secondly, amyloidogenic sequences that are found in beta structure are usually buried within the protein. Surface exposed amyloidogenic sequences are not tolerated in strands, presumably because they lead to protein aggregation via assembly of the amyloidogenic regions. The use of alpha-helices, where amyloidogenic sequences are forced into helix, despite their intrinsic preference for beta structure, is thus a widespread mechanism to avoid protein aggregation. PMID- 20027622 TI - The startle response: developmental effects and a paradigm for children and adults. AB - A film paradigm was developed to examine baseline and emotion modulated startle across a broad age range from preschool to adulthood. The paradigm was tested in children (3-, 5-, 7-, and 9-year-olds) and adults (total N = 122). The paradigm elicited a similar startle potentiation pattern across age groups; however, baseline startle changed with age: 3- and 5-year-olds showed lower response probability and magnitude of baseline startle than adults. Females exhibited larger baseline startle response probability and overall magnitude than did males; however, no sex by emotion modulated startle interaction was noted. Anxiety measures were obtained for all children. Individual differences in anxiety were associated with baseline startle magnitude among older but not younger children. No association of anxiety with startle potentiation was noted. Overall the film paradigm was applicable across a wide developmental span, revealing potential developmental and gender differences in baseline startle magnitude and response probability. PMID- 20027623 TI - Integrated discrimination improvement and probability-sensitive AUC variants. PMID- 20027624 TI - Mobile loop mutations in an archaeal inositol monophosphatase: modulating three metal ion assisted catalysis and lithium inhibition. AB - The inositol monophosphatase (IMPase) enzyme from the hyperthermophilic archaeon Methanocaldococcus jannaschii requires Mg(2+) for activity and binds three to four ions tightly in the absence of ligands: K(D) = 0.8 muM for one ion with a K(D) of 38 muM for the other Mg(2+) ions. However, the enzyme requires 5-10 mM Mg(2+) for optimum catalysis, suggesting substrate alters the metal ion affinity. In crystal structures of this archaeal IMPase with products, one of the three metal ions is coordinated by only one protein contact, Asp38. The importance of this and three other acidic residues in a mobile loop that approaches the active site was probed with mutational studies. Only D38A exhibited an increased kinetic K(D) for Mg(2+); D26A, E39A, and E41A showed no significant change in the Mg(2+) requirement for optimal activity. D38A also showed an increased K(m), but little effect on k(cat). This behavior is consistent with this side chain coordinating the third metal ion in the substrate complex, but with sufficient flexibility in the loop such that other acidic residues could position the Mg(2+) in the active site in the absence of Asp38. While lithium ion inhibition of the archaeal IMPase is very poor (IC(50) approximately 250 mM), the D38A enzyme has a dramatically enhanced sensitivity to Li(+) with an IC(50) of 12 mM. These results constitute additional evidence for three metal ion assisted catalysis with substrate and product binding reducing affinity of the third necessary metal ion. They also suggest a specific mode of action for lithium inhibition in the IMPase superfamily. PMID- 20027625 TI - Crystal structure of a super leucine zipper, an extended two-stranded super long coiled coil. AB - Coiled coil is a ubiquitous structural motif in proteins, with two to seven alpha helices coiled together like the strands of a rope, and coiled coil folding and assembly is not completely understood. A GCN4 leucine zipper mutant with four mutations of K3A, D7A, Y17W, and H18N has been designed, and the crystal structure has been determined at 1.6 A resolution. The peptide monomer shows a helix trunk with short curved N- and C-termini. In the crystal, two monomers cross in 35 degrees and form an X-shaped dimer, and each X-shaped dimer is welded into the next one through sticky hydrophobic ends, thus forming an extended two stranded, parallel, super long coiled coil rather than a discrete, two-helix coiled coil of the wild-type GCN4 leucine zipper. Leucine residues appear at every seventh position in the super long coiled coil, suggesting that it is an extended super leucine zipper. Compared to the wild-type leucine zipper, the N terminus of the mutant has a dramatic conformational change and the C-terminus has one more residue Glu 32 determined. The mutant X-shaped dimer has a large crossing angle of 35 degrees instead of 18 degrees in the wild-type dimer. The results show a novel assembly mode and oligomeric state of coiled coil, and demonstrate that mutations may affect folding and assembly of the overall coiled coil. Analysis of the formation mechanism of the super long coiled coil may help understand and design self-assembling protein fibers. PMID- 20027626 TI - A novel defensin-like peptide from salivary glands of the hard tick, Haemaphysalis longicornis. AB - A novel defensin-like antimicrobial peptide named longicornsin was isolated from the salivary glands of the hard tick, Haemaphysalis longicornis, using a 10-kDa cut-off Centriprep filter and reversed-phase high-performance liquid chromatography (RP-HPLC). Its amino acid sequence was determined as DFGCGQGMIFMCQRRCMRLYPGSTGFCRGFRCMCDTHIPLRPPFMVG by Edman degradation. The cDNA encoding longicornsin was cloned by cDNA library screening. The predicted protein from the cDNA sequence was composed of 78 amino acids including a mature longicornsin. It showed similarity with defensin-like peptides from other ticks by BLAST search. Different from most other tick defensin-like peptides, longicornsin had a C-terminal extension. Purified longicornsin exerted potent antimicrobial activities against bacteria and fungi. Interestingly, it even showed strong antimicrobial ability against drug-resistant microorganisms and Helicobacter pylori. The results of this study indicated that longicornsin is a potential candidate for novel antimicrobial drug design. PMID- 20027627 TI - Bereavement care: some clinical observations. AB - The needs of cancer patients' bereaved carers require attention. Although the majority of bereaved individuals manage to cope with their loss, a minority remain distressed and suffer serious prolonged disruption of their lives. The Yale Bereavement Study has revealed a specific syndrome, prolonged grief disorder. The diagnostic criteria of this disorder are outlined and several clinical illustrations are presented. Preliminary evidence suggests that cognitive-behaviour therapy (CBT) can be effective in the management of prolonged grief disorder. An example of the use of CBT is provided together with some useful clinical guidelines. PMID- 20027628 TI - MMP-2 alters VEGF expression via alphaVbeta3 integrin-mediated PI3K/AKT signaling in A549 lung cancer cells. AB - Vascular endothelial growth factor (VEGF) is one of the most important angiogenic growth factors for tumor angiogenesis. Here, we sought to explore whether RNA interference (RNAi) targeting matrix metalloproteinase-2 (MMP-2) could disrupt VEGF-mediated angiogenesis in lung cancer. MMP-2 siRNA inhibited lung cancer cell induced tube formation of endothelial cells in vitro; addition of recombinant human-MMP-2 restored angiogenesis. MMP-2 transcriptional suppression decreased VEGF, phosphatidylinositol 3-kinase (PI3K) protein levels and AKT phosphorylation in lung cancer cells. In addition, MMP-2 suppression decreased hypoxia inducible factor-1alpha (HIF-1alpha), a transcription factor for VEGF, as determined by electrophoretic mobility shift assay (EMSA). We also show that MMP-2 suppression disrupted PI3K dependent VEGF expression; ectopic expression of myr-AKT restored VEGF inhibition. Further, MMP-2 suppression decreased the interaction of integrin alphaVbeta3 and MMP-2 as confirmed by immunoprecipitation analyses. Studies with either function blocking integrin-alphaVbeta3 antibody or MMP-2 specific inhibitor (ARP-100) indicate that suppression of MMP-2 decreased integrin alphaVbeta3-mediated induction of PI3K/AKT leading to decreased VEGF expression. Moreover, A549 xenograft tissue sections from mice that treated with MMP-2 siRNA showed reduced expression of VEGF and the angiogenic marker, factor-VIII. The inhibition of tumor angiogenesis in MMP-2 suppressed tumor sections was associated with decreased co-localization of integrin-alphaVbeta3 and MMP-2. In summary, these data provide new insights into the mechanisms underlying MMP-2 mediated VEGF expression in lung tumor angiogenesis. PMID- 20027629 TI - Nucleophosmin is recognized by a cytotoxic T cell line derived from a rectal carcinoma patient. AB - Immunotherapy of colorectal carcinoma (CRC) has great promise as the presence of T lymphocytes in CRC tissues in situ is correlated with reduced recurrence and increased survival. Thus, identification of the antigens recognized by T cells of CRC patients may permit development of vaccines with potential benefit for these patients. Using expression cloning, we identified the antigen, nucleophosmin (Npm), recognized by an HLA-A1 restricted cytotoxic T lymphocyte (CTL) line derived from the peripheral blood mononuclear cells (PBMC) of a rectal cancer patient. A decamer peptide derived from the Npm sequence sensitized peptide pulsed HLA-A1 positive cells to lysis by the CTL line. The peptide also induced proliferative and cytotoxic T lymphocytes in the PBMC of 4 of 6 CRC patients, which lysed HLA-A1 positive peptide-pulsed target cells and CRC cells endogenously expressing Npm. Overexpression of Npm by tumors of various histological types, recognition of the antigen by T cells derived from different CRC patients and association of the antigen with poor prognostic outcome make it a promising target for immunotherapeutic intervention in cancer patients. PMID- 20027630 TI - Human monoclonal antibody GNX-8 directed to extended type 1 chain: Specific binding to human colorectal cancer. AB - We observed previously that two carbohydrate epitopes, extended type 1 chain Le(a)-Le(a) and Le(b)-Le(a), are expressed strongly in human gastric or colorectal cancer and cell lines derived therefrom, but their expression in human normal colorectal cells is highly limited. A monoclonal antibody, termed GNX-8, was established through immunization of "KM mice" with colonic cancer cell line Colo205, and with purified Le(b)-Le(a) glycosphingolipid, followed by screening human IgG directed to this antigen. KM mice possess human chromosome fragments and are capable of producing human immunoglobulin. GNX-8 reacted specifically with extended type 1 chain epitope Le(b)-Le(a), bound to all five colonic cancer cell lines so far tested, and displayed strong complement-dependent cytotoxicity (CDC) and antibody-dependent cellular cytotoxicity (ADCC). The antigens defined by GNX-8, expressed in Colo205 cells, were: (i) glycosphingolipids with epitope Le(b)-Le(a), whose reactivity was abolished upon defucosylation; (ii) glycoproteins with molecular mass range from 32 to >175 kDa, which were depleted in cells cultured in the presence of benzyl-alpha-GalNAc, indicating that these epitopes are O-linked glycans.Immunohistological reactivity of GNX-8 at 1 mug/ml, applied on tissue sections from colorectal and various other types of cancer, was much stronger than that with various normal cells and tissues. GNX-8 reactivity with normal cells required a much higher concentration (150 mug/ml), and this reactivity was based on cross-reaction with non-extended, normal blood group Le(b) antigen. Growth of subcutaneous xenograft of human colonic cancer cells, Colo205 or DLD-1, in nude mice or SCID mice, was strongly inhibited by administration of GNX-8. These observations, taken together, indicate that antibody GNX-8, directed specifically to Le(b)-Le(a) antigen, provides a novel direction of immunotherapy for human colorectal cancer. (c) 2009 UICC. PMID- 20027631 TI - Selective concomitant inhibition of mTORC1 and mTORC2 activity in estrogen receptor negative breast cancer cells by BN107 and oleanolic acid. AB - Hormonal, targeted and chemotherapeutic strategies largely depend on the expression of their cognate receptors and are often accompanied by intolerable toxicities. Effective and less toxic therapies for estrogen receptor negative (ER ) breast cancers are urgently needed. Here, we present the potential molecular mechanisms mediating the selective pro-apoptotic effect induced by BN107 and its principle terpene, oleanolic acid (OA), on ER- breast cancer cells. A panel of breast cancer cell lines was examined and the most significant cytotoxic effect was observed in ER- breast lines. Apoptosis was the major cellular pathway mediating the cytotoxicity of BN107. We demonstrated that sensitivity to BN107 was correlated to the status of ERalpha. Specifically, the presence of functional ERalpha protected cells from BN107-induced apoptosis and absence of ERalpha increased the sensitivity. BN107, an extract rich in OA derivatives, caused rapid alterations in cholesterol homeostasis, presumably by depleting cholesterol in lipid rafts (LRs), which subsequently interfered with signaling mediated by LRs. We showed that BN107 or OA treatment in ER- breast cancer cells resulted in rapid and specific inhibition of LR-mediated survival signaling, namely mTORC1 and mTORC2 activities, by decreasing the levels of the mTOR/FRAP1, RAPTOR and RICTOR. Cotreatment with cholesterol abolished the proapoptotic effect and restored the disrupted mTOR activities. This is the first report demonstrating possible concomitant inhibition of both mTORC1 and mTORC2 activities by modulating the levels of protein constituents present in these signaling complexes, and thus provides a basis for future development of OA-based mTOR inhibitors. PMID- 20027632 TI - Regulatory T cells control macrophage accumulation and activation in lymphoma. AB - Strategies of manipulating immunosuppressive regulatory T cells (Treg) in cancer patients are currently evaluated in clinical trials. Treg suppress immune responses of tumor-specific T cells; yet, relatively little is known about the impact of Treg on innate immune cells in tumor models in vivo. Many tumors lose expression of MHC class I. Therefore, our study aimed at defining strategies to strengthen immune responses against a high tumor burden of the MHC class I deficient mouse lymphoma RMA-S. We demonstrate that Treg depletion in mice led to tumor rejection that was dependent on T cells, NK cells and IFN-gamma. In the absence of Treg elevated levels of IFN-gamma were produced by tumor-infiltrating T cells and NK cells. Tumor rejection observed in the absence of Treg correlated with a substantial IFN-gamma-dependent increase in the numbers of tumor infiltrating leukocytes. The most abundant cell population in the tumors was macrophages. Tumor-infiltrating macrophages from Treg-depleted mice expressed increased amounts of MHC class II, produced highly enhanced levels of pro inflammatory cytokines and inhibited tumor cell proliferation. It was reported that tumor-infiltrating macrophages have multi-faceted functions promoting or counteracting tumor growth. In our study, high numbers of macrophages infiltrating RMA-S tumors in the absence of Treg correlated with tumor rejection suggesting that macrophages are additional targets for Treg-mediated immune suppression in cancer. PMID- 20027633 TI - Granzyme B is expressed in urothelial carcinoma and promotes cancer cell invasion. AB - Granzyme B (GrB) is a serine proteinase known to be expressed by cytotoxic lymphocytes and to induce, in presence of perforin (Pf), apoptosis in target cells. Recently, GrB expression has been shown (often in absence of Pf) in nonlymphoid cells, but its function is not defined. In our study, we investigated GrB and Pf expression in bladder cancer cell lines and in urothelial carcinoma (UC) tissues by reverse transcription-polymerase chain reaction (RT-PCR), Western blot, ELISA, immunofluorescence and immunohistochemistry. We also assessed the function of GrB in UC cells; the in vitro function of GrB was examined by loss-of function experiments. Our results revealed that GrB is expressed, in absence of Pf, in UC cells. Significant differences were found between GrB expression and both increasing pathological tumor spreading and high-grade vs. low-grade pTa tumors. Notably, GrB in UC tissues was concentrated at the cancer invasion front and was expressed in neoplastic cells undergoing epithelial-mesenchymal transition, a key event in carcinoma invasion. Indeed, GrB-positive cells also expressed Snail, N-cadherin or were negative for E-cadherin. GrB expressed in tumor cell lines was enzymatically active and capable of vitronectin cleavage, implying extracellular matrix (ECM) remodeling by GrB. Inhibition of GrB activity or Stealth RNA interference-mediated GrB gene silencing markedly suppressed bladder cancer cell invasion through matrigel. This data provides the first evidence for a role of GrB in promoting cancer cell invasion. Taken together, our findings suggest that GrB, via ECM degradation, contributes to the establishment of the UC invasive phenotype. PMID- 20027634 TI - Sexually dimorphic tegumental gland openings in Laniatores (Arachnida, Opiliones), with new data on 23 species. AB - Sexually dimorphic glands often release sexual pheromones both in vertebrates and invertebrates. Species of Laniatores (Arachnida, Opiliones) seem to depend on chemical communication but few studies have addressed this topic. In this study, we review the literature for the Phalangida and present new data for 23 species of Laniatores. In 16 taxa, we found previously undescribed sexually dimorphic glandular openings on the femur, patella, metatarsus, and tarsus of legs I and metatarsus of legs III and IV. For the other species, we provide scanning electron micrographs of previously undescribed sexually dimorphic setae and pegs located on swollen regions of the legs. We also list additional species in which males have swollen regions on the legs, including the tibia, metatarsus, and tarsus of legs I, trochanter and tibia of legs II, femur, metatarsus, and tarsus of legs III, and metatarsus and tarsus of legs IV. The function and biological role of the secretions released by these glands are discussed. PMID- 20027635 TI - PROMO: Real-time prospective motion correction in MRI using image-based tracking. AB - Artifacts caused by patient motion during scanning remain a serious problem in most MRI applications. The prospective motion correction technique attempts to address this problem at its source by keeping the measurement coordinate system fixed with respect to the patient throughout the entire scan process. In this study, a new image-based approach for prospective motion correction is described, which utilizes three orthogonal two-dimensional spiral navigator acquisitions, along with a flexible image-based tracking method based on the extended Kalman filter algorithm for online motion measurement. The spiral navigator/extended Kalman filter framework offers the advantages of image-domain tracking within patient-specific regions-of-interest and reduced sensitivity to off-resonance induced corruption of rigid-body motion estimates. The performance of the method was tested using offline computer simulations and online in vivo head motion experiments. In vivo validation results covering a broad range of staged head motions indicate a steady-state error of less than 10% of the motion magnitude, even for large compound motions that included rotations over 15 deg. A preliminary in vivo application in three-dimensional inversion recovery spoiled gradient echo (IR-SPGR) and three-dimensional fast spin echo (FSE) sequences demonstrates the effectiveness of the spiral navigator/extended Kalman filter framework for correcting three-dimensional rigid-body head motion artifacts prospectively in high-resolution three-dimensional MRI scans. PMID- 20027636 TI - Stat3 is required for anchorage-independent growth and metastasis but not for mammary tumor development downstream of the ErbB-2 oncogene. AB - The oncogenic transcription factor Stat3 is constitutively active in a high percentage of human tumors including mammary adenocarcinomas and is reported to participate in the ErbB-2 oncogene signaling. In order to assess the role of signal transducer and activator of transcription 3 (Stat3) in mammary tumorigenesis downstream of ErbB-2, we generated mice expressing the activated rat ErbB-2 (neu) but lacking Stat3 in the mammary epithelium. Stat3 is apparently not required for neu-driven mammary tumorigenesis as tumors developed similarly in both Stat3-sufficient and Stat3-deficient glands. However, short hairpin RNA (shRNA)-mediated Stat3 silencing in a neu-overexpressing tumor-derived cell line completely abolished both neu-driven anchorage-independent growth and lung metastasis. Our data suggest that Stat3 might be a useful therapeutic target in breast tumors showing amplification and/or overexpression of the ErbB-2 oncogene, which normally display aggressive, metastatic behavior. PMID- 20027637 TI - Seasonal and reproductive variation in body condition in captive female Japanese macaques (Macaca fuscata). AB - The geographic distribution of Japanese macaques includes populations with the most northern range of any primate species. Not surprisingly, females of this species are characterized by physiological adaptations and unique fat deposition mechanisms that facilitate their survival through the sometimes-harsh seasonal conditions of temperate climates, as well as sustaining the metabolic costs of mating, pregnancy, and lactation. Here, we explore the relationship between nutritional status, seasonality, and reproductive status using anthropometric and leptin measures from 14 captive female Japanese macaques. No seasonal patterns were found in the levels of leptin, but there were differences between seasons in anthropometric measures, specifically between the beginning and the end of the mating season. Females gained weight and accumulated energy reserves in fall to prepare for mating activity, and to survive the severe conditions of winter, which is also the period of gestation if pregnancy occurs. Lactating females had larger total skinfolds relative to nonlactating individuals, and females with older babies at the beginning of the mating season had larger abdominal skinfolds than did those with younger babies. There was a relationship between the likelihood of conception and nutritional status, with females that conceived during one mating season being in better condition at the end of their previous mating season. Together, these results suggest that, even in captive settings, seasonal breeding has a cost on the energetic demands of mating, and that higher condition (i.e. fatter) females could afford the demands of lactation and reproduced more rapidly. PMID- 20027638 TI - Association between activation of atypical NF-kappaB1 p105 signaling pathway and nuclear beta-catenin accumulation in colorectal carcinoma. AB - Recent studies have demonstrated that increased expression of coding region determinant-binding protein (CRD-BP) in response to beta-catenin signaling leads to the stabilization of beta-TrCP1, a substrate-specific component of SCF E3 ubiquitin ligase complex, resulting in an accelerated degradation of IkappaBalpha and activation of canonical nuclear factor-kappaB (NF-kappaB) pathway. Here, we show that the noncanonical NF-kappaB1 p105 pathway is constitutively activated in colorectal carcinoma specimens, being particularly associated with beta-catenin mediated increased expression of CRD-BP and beta-TrCP1. In the carcinoma tissues exhibiting high levels of nuclear beta-catenin the phospho-p105 levels were increased and total p105 amounts were decreased in comparison to that of normal tissue indicating an activation of this NF-kappaB pathway. Knockdown of CRD-BP in colorectal cancer cell line SW620 resulted in significantly higher basal levels of both NF-kappaB inhibitory proteins, p105 and IkappaBalpha. Furthermore decreased NF-kappaB binding activity was observed in CRD-BP siRNA-transfected SW620 cells as compared with those transfected with control siRNA. Altogether, our findings suggest that activation of NF-kappaB1 p105 signaling in colorectal carcinoma might be attributed to beta-catenin-mediated induction of CRD-BP and beta-TrCP1. PMID- 20027639 TI - Diet of the Delacour's langur (Trachypithecus delacouri) in Van Long Nature Reserve, Vietnam. AB - The six limestone langur taxa of Southeast Asia inhabit the rugged limestone karst mountains, although the reason for their current restriction to this habitat is unclear. From August 2007 to July 2008, I collected data on the diet and feeding behavior of the critically endangered Delacour's langur (Trachypithecus delacouri) in Van Long Nature Reserve, Vietnam. I used these data to evaluate the hypothesis that limestone langurs are found on karst habitat because they depend on endemic limestone plants. Feeding accounted for 29% of the langurs' activity budget. Young leaves dominated the diet monthly, seasonally, and annually. The annual diet consists of nearly 79% foliage with almost 60% young leaves. Despite a distinct wet and dry season, over the study period, seasonal variation in plant part consumption was slight. Fruit and seeds were a small contribution to the diet. Delacour's langurs ate 42 of 145 available species, and they concentrated on a subset of this number. Five plant species comprised more than 60% of the diet and 16 species comprised more than 93%. More than half of the diet came from climbers. Delacour's langurs are among the most highly folivorous of studied colobines and, along with the closely related T. leucocephalus of southern China, the most folivorous of the Asian langurs. Whether high folivory is owing to a lack of available fruits and seeds in limestone habitats is unknown. What is certain, however, is that the plant species most important in the Delacour's langurs' diet at VLNR, throughout the study, were not plants endemic to limestone. Feeding dependence alone, therefore, cannot explain the current distribution of limestone langurs on karst habitat. PMID- 20027640 TI - Prevention of Rh sensitization in the context of trauma: two case reports. AB - BACKGROUND: Transfusion of D+ red blood cells (RBCs) to D- recipients can be accidental or necessary due to D- RBC shortage. Alloimmunization can complicate future transfusions; implications for women of childbearing age are compounded by possible hemolytic disease of the fetus and newborn. Rh immunoprophylaxis is effective, and indicated, for preventing alloimmunization. Reports of massive D+ mismatch (e.g., in the case of fetal-maternal bleed) are limited, and standard recommendations for managing these rare events are lacking. The cases discussed herein of women of childbearing age who suffered severe trauma requiring emergency surgery illustrate the dilemma of determining the ideal strategy for Rh immunoprophylaxis. CASE REPORTS: The first patient received two units of mismatched RBCs and was treated with intravenous Rh immune globulin (IV RHIG; Rhophylac, CSL Behring, Kankakee, IL) monotherapy beginning 49 h postsurgery. For the second patient, who received three units of D+ RBCs, partial RBC exchange transfusion, followed 48 h later by IV RHIG, was deemed appropriate based on the large volume of RHIG needed and concerns of hemolytic transfusion reaction and hyperbilirubinemia. Both patients recovered in full without further intervention; the first patient delivered a healthy child approximately 11 months posttreatment. CONCLUSIONS: Rh immunoprophylaxis is effective, and indicated, for preventing alloimmunization in women of childbearing age to protect the mother, fetus, and newborn. For the general population, a case-by-case approach is recommended. The availability of IV RHIG improves patient safety by facilitating the delivery of such treatment. RBC exchange should be considered for large volume mismatches. PMID- 20027641 TI - The QM-MM interface for CHARMM-deMon. AB - We present a new QM/MM interface for fast and efficient simulations of organic and biological molecules. The CHARMM/deMon interface has been developed and tested to perform minimization and atomistic simulations for multi-particle systems. The current features of this QM/MM interface include readability for molecular dynamics, tested compatibility with Free Energy Perturbation simulations (FEP) using the dual topology/single coordinate method. The current coupling scheme uses link atoms, but further extensions of the code to incorporate other available schemes are planned. We report the performance of different levels of theory for the treatment of the QM region, while the MM region was represented by a classical force-field (CHARMM27) or a polarizable force-field based on a simple Drude model. The current QM/MM implementation can be coupled to the dual-thermostat method and the VV2 integrator to run molecular dynamics simulations. PMID- 20027642 TI - A discriminative approach for identifying domain-domain interactions from protein protein interactions. AB - Protein domains are functional and structural units of proteins. Therefore, identification of domain-domain interactions (DDIs) can provide insight into the biological functions of proteins. In this article, we propose a novel discriminative approach for predicting DDIs based on both protein-protein interactions (PPIs) and the derived information of non-PPIs. We make a threefold contribution to the work in this area. First, we take into account non-PPIs explicitly and treat the domain combinations that can discriminate PPIs from non PPIs as putative DDIs. Second, DDI identification is formalized as a feature selection problem, in which it tries to find out a minimum set of informative features (i.e., putative DDIs) that discriminate PPIs from non-PPIs, which is plausible in biology and is able to predict DDIs in a systematic and accurate manner. Third, multidomain combinations including two-domain combinations are taken into account in the proposed method, where multidomain cooperations may help proteins to interact with each other. Numerical results on several DDI prediction benchmark data sets show that the proposed discriminative method performs comparably well with other top algorithms with respect to overall performance, and outperforms other methods in terms of precision. The PPI data sets used for prediction of DDIs and prediction results can be found at http://csb.shu.edu.cn/dipd. PMID- 20027643 TI - Neonatal gonocyte differentiation in Mongolian gerbil Meriones unguiculatus involves asynchronous maturation of seminiferous cords and rapid formation of transitional cell stage. AB - This study describes the neonatal differentiation of the Mongolian gerbil gonocytes, focusing on the relationship between its relocation to the basement membrane, apoptosis and postrelocation changes and also the distribution of androgen receptors (AR). Testes of gerbils from 1 to 35 days of age (d) were examined by high resolution light microscopy and immunocytochemistry for proteins PCNA, VASA, and AR as well as by the TUNEL method. Gonocytes were quantified according to degree of relocation into nonrelocated, relocating and relocated. Most of them were found in the center of seminiferous cords at 1 d but a small number of relocating and relocated gonocytes were already visible in the first postnatal day. After relocation, gonocytes change phenotypically to a transitional stage designated herein prospermatogonia. Both gonocyte relocation and transformation into spermatogonial lineage occur asynchronously in the seminiferous cords, mainly after 7 d. Gonocyte proliferation began before but peak after their relocation to basement membrane at the prospermatogonia stage. Higher levels of gonocyte apoptosis were found at 7 d and 21 d. From this time onward gonocytes were not found. Gonocytes and prospermatogonia showed high amounts of AR in their cytoplasm contrary to spermatogonial subtypes, indicating a possible AR inactivation in these cells. In conclusion, the process of gonocyte relocation in the gerbil extends until the second postnatal week, leads to their rapid differentiation into prospermatogonia and occurs simultaneously with the loss of androgen sensitivity. Differently from other laboratory rodents, the events regarding gonocyte maturation in the gerbil last longer and occur asynchronously in seminiferous cords. PMID- 20027644 TI - Genetically engineered mesenchymal stem cells: The ongoing research for bone tissue engineering. AB - Bone grafting is crucial in the surgical treatment of bone defects and nonunion fractures. Autogenous bone, allogenous bone, and biomaterial scaffold are three main sources of bone grafts. The biomaterial scaffold, both natural and synthetic, is widely accessible but weak in osteogenic potential. One approach to solve this problem is cell-based bone tissue engineering (BTE), established by growing living osteogenic cells on scaffold in vitro to build up its osteoinducitive capability. Mesenchymal stem cell (MSC) is suitable for use in cell-based BTE, but it remains a considerable challenge to induce MSCs to form solely bone and while preventing MSCs from differentiating into fats, muscles, and possibly neural elements in vivo. Recently, there is a drastic rise in use of genetically engineered MSCs, which can secrete growth factors or alter the transcription level, leading to osteoblast lineage commitment, bone formation, fracture repair, and spinal fusion. In this article, we reviewed the literatures regarding applications of genetically engineered MSCs in BTE. We addressed the currently applicable genes and candidate genes for MSCs modification, transduction efficiency and safety issues of the transfect vectors, and administration routes, and we briefly described in vivo tracking and potential clinical application of the genetically modified MSCs in BTE. PMID- 20027645 TI - Bone density distribution patterns in the rostrum of delphinids and beaked whales: evidence of family-specific evolutive traits. AB - Toothed whales have undergone a profound telescopic rearrangement of the skull, with elongation of facial bones and formation of a hollow rostrum, filled in vivo by the mesorostral cartilage. In most species of the family Ziphiidae, this latter cartilage becomes secondarily ossified, producing in some cases the densest bone existing in nature. Starting from this observation, we wanted to investigate the patterns of distribution of bone mineral density (BMD) in the rostrum of two families of toothed whales with different ecological and behavioral traits: Delphinidae and Ziphiidae. We analyzed BMD non invasively by means of the dual energy X-ray absorptiometry technology, and found two different density distribution patterns that distinctly set the two families apart. Namely, BMD values decrease from the proximal to the distal region of the rostrum in delphinids, whereas the beaked whales show a BMD peak in the central region. Possible functions such as ballast or protection against clashes might be likely, although more data about the species of both families is needed to give better evidence. PMID- 20027646 TI - Piezoresistive cantilever array sensor for consolidated bioprocess monitoring. AB - Cellulolytic microbes occur in diverse natural niches and are being screened for industrial modification and utility. A microbe for consolidated bioprocessing (CBP) development can rapidly degrade pure cellulose and then ferment the resulting sugars into fuels. To identify and screen for novel microbes for CBP, we have developed a piezoresistive cantilever array sensor which is capable of simultaneous monitoring of glucose and ethanol concentration changes in a phosphate buffer solution. 4-mercaptophenylboronic acid and polyethyleneglycol thiol are employed to functionalize each piezoresistive cantilever for glucose and ethanol sensing, respectively. Successful concentration measurements of glucose and ethanol with minimal interferences are obtained with our cantilever array sensor. PMID- 20027647 TI - Safety of adalimumab in Crohn's disease during pregnancy: case report and review of the literature. PMID- 20027648 TI - When expression is not enough: the evolving thoughts on Fas/FasL and epithelial cell apoptosis. PMID- 20027649 TI - Should patients under long-term anti-TNF therapies be followed for tuberculosis contamination? PMID- 20027650 TI - Combination of innate and adaptive immune alterations increased the likelihood of fibrostenosis in Crohn's disease. AB - BACKGROUND: Mutations in the nucleotide oligomerization domain-2 (NOD2) gene and positive antibodies to microbial antigens have been found to be associated with the Crohn's disease (CD) phenotype, fibrostenosis. The aim of this study was to confirm these relationships in a large cohort of CD patients and to determine the correlation between the presence of NOD2 variants and antibodies to oligomannan, CBir, outer membrane porin-C (OmpC), and I2 in CD patients with fibrostenosis. METHODS: Sera and DNA from 731 unrelated CD patients were tested for NOD2 variants (SNP 8, 12, and 13) and the antibodies. The results were correlated with CD phenotypes, fibrostenosis, internal penetrating, perianal penetrating, and ulcerative colitis (UC)-like as well as other clinical features. RESULTS: The presence of NOD2 allelic variants was primarily associated with fibrostenosis, secondarily with small bowel disease and small bowel surgery, and was inversely associated with UC-like disease. This association was present in patients with a fibrostenosis only (Vienna B2) and those with both stricturing and penetrating disease. The presence and level of antibodies to microbial antigens was also associated with the fibrostenosis phenotype. In the 316 patients with fibrostenosis the prevalence of NOD2 variants was significantly correlated with the antibody titer by quartile sum score. Further, when these patients with fibrostenosis were clustered by quartile sum score, the odds ratio for fibrostenosis was significantly higher in the patients with NOD2 variant alleles within each cluster, indicating synergy. CONCLUSIONS: Defects of innate (NOD2 variants) and adaptive (antibodies to microbial antigens) immunity act synergistically to increase the risk of the fibrostenosis phenotype. PMID- 20027651 TI - Human embryonic stem cell lines and their use in international research. AB - Research in human pluripotent stem cells, including human embryonic stem cells (hESC) and human induced pluripotent stem cells (hiPSC), is one of the most dynamic research fields. Despite the high public attention, especially for hESC research, there is only scattered information on the number of hESC lines and the degree, dynamics, and diversification of their use on a global level. In this study we present data on the current number of publicly disclosed hESC lines, on the extent and impact of experimental work involving hESCs, and on the use of specific hESC lines in international research. The results are based on the evaluation of nearly 1,000 research papers published by the end of 2008, which describe experimental work on hESCs, and of a comprehensive database of published hESC lines. The average impact of hESC research papers is high at 7.422, with a predominance of research output by the United States. Of at least 1,071 original hESC lines derived up to November 2009 at 87 institutions in 24 countries, only a fraction is thoroughly characterized. Our data show the global predominance of a few hESC lines in research, but also reveal remarkable country-specific differences. Comparison of hESC and hiPSC application did not show a diminished role for hESC research, but rather revealed that, up to this time, both fields continue to expand, exist independently, and partially overlap. PMID- 20027652 TI - Mortality in inflammatory bowel disease in the Netherlands 1991-2002: results of a population-based study: the IBD South-Limburg cohort. AB - BACKGROUND: The aim was to evaluate overall and disease-specific mortality in a population-based inflammatory bowel disease (IBD) cohort in the Netherlands, as well as risk factors for mortality. METHODS: IBD patients diagnosed between 1 January 1991 and 1 January 2003 were included. Standardized mortality ratios (SMRs) were calculated overall and with regard to causes of death, gender, as well as age, phenotype, smoking status at diagnosis, and medication use. RESULTS: At the censoring date, 72 out of 1187 patients had died (21 Crohn's disease [CD], 47 ulcerative colitis [UC], and 4 indeterminate colitis [IC] patients). The SMR (95% confidence interval [CI]) was 1.1 (0.7-1.6) for CD, 0.9 (0.7-1.2) for UC and 0.7 (0.2-1.7) for IC. Disease-specific mortality risk was significantly increased for gastrointestinal (GI) causes of death both in CD (SMR 7.5, 95% CI: 2.8-16.4) and UC (SMR 3.4, 95% CI: 1.4-7.0); in CD patients, especially in patients <40 years of age at diagnosis. For UC, an increased SMR was noted in female patients and in patients <19 years and >80 years at diagnosis. In contrast, UC patients had a decreased mortality risk from cancer (SMR 0.5, 95% CI; 0.2-0.9). CONCLUSIONS: In this population-based IBD study, mortality in CD, UC, and IC was comparable to the background population. The increased mortality risk for GI causes might reflect complicated disease course, with young and elderly patients at diagnosis needing intensive follow-up. Caution in interpreting the finding on mortality risk from cancer is needed as follow-up was probably to short to observe IBD-related cancers. PMID- 20027654 TI - Budesonide treatment of patients with collagenous colitis restores normal eosinophil and T-cell activity in the colon. AB - BACKGROUND: The aim of this study was to assess the activity of eosinophils, neutrophils, and CD4+ as well as CD8+ T-cells in 11 patients with active collagenous colitis (CC) before and after 8 weeks of budesonide treatment (9 mg once daily) compared to 10 healthy individuals. METHODS: Clinical symptoms were recorded and intestinal biopsy samples were taken and analyzed by flow cytometry. Eosinophils with a high surface expression of CD44 and low CD9 expression were classified as activated. Neutrophil activity was assessed by their expression of CD66b, and CD69 was used as an activation marker for T-cells. RESULTS: All patients responded to the treatment. The eosinophils in active CC showed increased activity compared to controls. The activity was back to control levels after treatment. Neutrophils were not activated in CC patients before or after treatment. CD8+ T-cells from untreated CC patients had a lower activity than controls, and a tendency of lower activity was observed on CD4+ T-cells. After treatment, the activity was increased on both types of T-cells and was not different from controls. CONCLUSIONS: In the present study we demonstrated that the inflammation in CC is characterized by activated eosinophils but there is no neutrophil activity. CD4+ and CD8+ T-cells are increased in numbers in active CC but, surprisingly, they had a lower grade of activity than in control subjects. The major finding of this study is that budesonide treatment restores the normal activation of eosinophils and T-cells, accompanied by clinical remission. PMID- 20027655 TI - Laparoscopic surgery for recurrent ileocolic Crohn's disease. AB - BACKGROUND: Laparoscopic (LAP) surgery is increasingly performed for primary ileocolic Crohn's disease (CD), but its application in patients with recurrent ileocolic CD is less well described. Our aim was to assess whether or not a laparoscopic approach was safe, feasible, and conferred meaningful short-term benefits in this patient population. METHODS: Patients undergoing LAP surgery for recurrent ileocolic CD at our institution from 1998-2008 were identified using a prospectively maintained database. Potential risk factors for conversion to open surgery and overall patient outcomes were assessed with univariate analysis. RESULTS: Forty patients were identified, of which 30 (75%) were LAP-completed and 10 (25%) were LAP-converted. The groups did not differ with respect to clinicopathological features. LAP-converted patients were significantly more likely to require adhesiolysis than LAP-completed patients (100% versus 67%, P = 0.04). There was 1 intraoperative complication in a converted patient. LAP converted patients had longer times to soft diet (4 versus 3 days, P = 0.03) and longer length of stay (7 versus 4 days, P = 0.003). The groups did not differ with respect to incidence of postoperative complications or frequency of readmission within 30 days. There was no mortality. CONCLUSIONS: In up to 20% of patients with recurrent ileocolic, successful laparoscopic re-resection may be prevented by adhesions. Conversion increased the length of stay without increasing morbidity. We conclude that LAP surgery can be safely performed in selected patients with recurrent ileocolic CD and leads to short-term benefits. PMID- 20027656 TI - Outcome after surveillance of low-grade and indefinite dysplasia in patients with ulcerative colitis. AB - BACKGROUND: The management of low-grade (LGD) and indefinite dysplasia (IND) in patients with ulcerative colitis (UC) remains controversial, as outcomes after a diagnosis of LGD or IND in previous studies vary widely. METHODS: All patients evaluated were from a single institution referral center who had a history of UC and a diagnosis of either LGD or IND between 1994 and 2008 as confirmed by 2 expert gastrointestinal (GI) pathologists. Data were collected by chart review of electronic and paper medical records. All patients who did not undergo a colectomy within 90 days of their dysplasia diagnosis were included in the final analysis. Hazard ratios for risk factors as well as incidence rates and Kaplan Meier estimates were used to calculate the progression to high-grade dysplasia (HGD) or colorectal cancer (CRC). RESULTS: Thirty-five patients were included in the analysis, of whom 2 patients with IND and 2 patients with LGD developed HGD or CRC over a mean duration of 49.8 months. In total, the incident rate for advanced neoplasia for all patients was 2.7 cases of HGD or CRC per 100 person years at risk. For flat and polypoid LGD the incident rate of advanced neoplasia was 4.3 and 1.5 cases per 100 person-years at risk, respectively. Patients with primary sclerosing cholangitis (PSC) had an incident rate of 10.5 cases per 100 years of patient follow-up. CONCLUSIONS: We report a low rate of progression to HGD or CRC in patients who underwent surveillance for LGD or IND; polypoid dysplasia showed less risk of progression than flat dysplasia. PMID- 20027657 TI - Impact of chronic antithrombotic therapy on hospital course of patients with acute myocardial infarction. AB - BACKGROUND: Little is known about the influence of chronic antithrombotic therapy on treatment and clinical outcome in patients with acute ST-elevation myocardial infarction (STEMI). HYPOTHESIS: The purpose of this study was to investigate the hospital course of STEMI patients on antithrombotics. METHODS: We analyzed data of consecutive patients with STEMI, who were prospectively enrolled in the German Acute Coronary Syndromes registry between July 2000 and November 2002. Overall, 8224 patients were stratified into 3 groups: group 1 had no prior chronic antithrombotic medication (n = 6004), group 2 was on chronic acetylsalicylic acid (ASA) therapy (n = 2022), and group 3 was on chronic oral anticoagulation therapy (n = 198). RESULTS: Patients on antithrombotic medication were older and had a higher baseline risk profile. The rate of patients receiving early reperfusion (group 1: 74.6%, group 2: 61.2%, group 3: 52.0%) and guideline-adherent adjustment therapy was lower among patients on antithrombotics. Age and left bundle branch block were strong negative predictors for early reperfusion therapy in patients with prior antithrombotic treatment. Infarct size measured by peak creatine kinase level was lower in patients on antithrombotics. Hospital mortality (group 1: 8.0%, group 2: 12.8%, group 3: 16.2%) and major bleeding complications (group 1: 1.6%, group 2 2.0%, group 3 4.1%) were highest in patients on oral anticoagulants. However, after adjustment for confounding factors, prior ASA (odds ratio [OR]: 0.98, 95% confidence interval [CI]: 0.80 1.21) and oral anticoagulant treatment (OR: 1.06, 95% CI: 0.66-1.71) were not independent predictors for in-hospital death. CONCLUSIONS: Despite a higher risk profile, patients with STEMI on a chronic antithrombotic therapy were less likely to receive early reperfusion therapy. However, after adjustment, prior ASA or oral anticoagulant therapy was not associated with higher in-hospital mortality. PMID- 20027658 TI - Evaluation of left atrial function in physiological and pathological left ventricular myocardial hypertrophy by real-time tri-plane strain rate imaging. AB - BACKGROUND: We investigated the difference between left ventricular (LV) hypertrophy caused by primary hypertension and physiological LV hypertrophy in athletes as seen in left atrial (LA) function by real-time tri-plane strain rate imaging. HYPOTHESIS: A real-time tri-plane imaging technique using the same phase of the same cardiac circle was used to synchronously demonstrate the section of apical 4-chamber, 2-chamber, and apical left ventricle long axis. METHODS: We measured standard Doppler echocardiographic quantitative analysis and the strain rate peak values of each LA wall in the systolic phase, in the early stage of diastole, and in the advanced stage of diastole and made a comparison of these values. RESULTS: The alteration of configuration and function of the left atrium in hypertensive patients is an early sign of the myocardial damage caused by hypertension. Strain rate imaging could sensitively reflect LA function changes in the early stages of hypertension. While physiological, myocardial hypertrophy is a benign reaction, LA function is significantly different from that of hypertension. CONCLUSIONS: Real-time tri-plane strain rate imaging techniques could simultaneously analyze 3 sections, which shortens scanning time and depletes the influence of variations of different cardiac cycles on quantitative analysis of local myocardial segments of the left atrium. This would improve the comparability of myocardial movement of different segments so that we could more comprehensively and accurately evaluate the systolic and diastolic function of the left atrium in primary hypertension and physiological LV hypertrophy in athletes. PMID- 20027659 TI - Statin therapy decreases myocardial function as evaluated via strain imaging. AB - OBJECTIVES: The purpose of this study was to evaluate the effects of statin therapy on myocardial function as measured with echocardiography with tissue Doppler imaging (TDI) and strain imaging (SI) independent of its lipid-lowering effect. BACKGROUND: Statin use is known to improve outcomes in the primary and secondary prevention of ischemic heart disease, but their use is also associated with myopathy, muscle weakness and in rare cases, rhabdomyolysis. We sought to evaluate whether TDI and SI is able to identify changes in myocardial function associated with statin use. METHODS: Myocardial function was evaluated in 28 patients via echocardiography with TDI and SI. We identified 12 patients (5 females) without overt cardiovascular disease (including hypertension, smoking, and diabetes) that were on statin therapy and compared their echocardiographic findings with 16 (12 females) age, sex, and cholesterol-profile-matched controls. Tissue Doppler imaging parameters of diastolic (E(')/A(') and E/E(')) and systolic (S') function were measured. Regional systolic function was obtained by SI in 4-chamber, 2-chamber, long axis, and average global views. RESULTS: There was no significant difference in myocardial function as measured by Doppler and minor differences as measured via TDI among the 2 groups. There was significantly better function noted with SI in the control group vs the statin group in the 4 chamber (-19.05% +/- 2.45% vs -16.47% +/- 2.37% P = 0.009), 2-chamber (-20.30% +/ 2.66% vs -17.45% +/- 4.29% P = 0.03), long axis (-17.63% +/- 3.79% vs -13.83% +/ 3.74% P = 0.01), and average global (-19.0% +/- 2.07% vs -15.91% +/- 2.81% P = 0.004) views. CONCLUSION: Statin therapy is associated with decreased myocardial function as evaluated with SI. PMID- 20027660 TI - N-terminal prohormone brain natriuretic peptide-proBNP levels in ventricular arrhythmias in children. AB - BACKGROUND: Ventricular arrhythmias are the most common consequences of structural and functional heart diseases, but cases with no evident pathology are also observed. A parameter indicating asymptomatic circulatory failure could support decisions related to possible treatment of ventricular arrhythmias. HYPOTHESIS: The study objective was the evaluation of N-terminal prohormone brain natriuretic peptide (NT-proBNP) levels in children with ventricular arrhythmias and an attempt to determine if this parameter may be used for diagnosis and prognosis of ventricular arrhythmias. MATERIAL AND METHODS: The study population was comprised of 36 children age 5 to 17.5 years old with idiopathic ventricular arrhythmias (Group B) graded mild or potentially malignant; 29 patients with mild ventricular arrhythmias were included into Group B1; and 7 patients with potentially malignant cases into Group B2. In all the patients, NT-proBNP assays were performed. RESULTS: The NT-proBNP levels in Groups B, B1, B2 and the control group (Group K) were as follows: 41.5 +/- 15.1 pg/mL, 35.5 +/- 18.5 pg/mL, 66.3 +/- 24.9 pg/mL and 31.5 +/- 15.1 pg/mL, respectively. Between the groups with and without arrhythmias (Group B vs Group K), no statistically significant differences in NT-proBNP levels were found. However, markedly higher NT-proBNP levels were shown in the children with potentially malignant arrhythmias (Group B2) compared to the patients with mild arrhythmias (B1) and the control group (Group K). CONCLUSIONS: The level of NT-proBNP increases with the severity of ventricular arrhythmia. NT-proBNP assays can be helpful for diagnosing and grading the severity of ventricular arrhythmias. PMID- 20027661 TI - Is right ventricular outflow tract pacing superior to right ventricular apex pacing in patients with normal cardiac function? AB - BACKGROUND: Whether right ventricular outflow tract (RVOT) pacing is superior to right ventricular apex (RVA) pacing in terms of ventricular synchrony, cardiac function, and remodeling in patients with normal cardiac function is still unknown. HYPOTHESIS: Right ventricular outflow tract pacing is superior to RVA pacing in patients with normal cardiac function. METHODS: A total of 96 consecutive patients with high or third-degree atrial ventricular block were enrolled and randomized into 2 groups: RVOT pacing group (n = 48) and RVA pacing group (n = 48). Tissue Doppler imaging (TDI) and 2D echocardiography were performed to study left ventricular (LV) systolic and diastolic synchrony, LV volumes, and function. RESULTS: There were no significant differences in baseline characteristics between the 2 groups. Left ventricular systolic asynchrony is more severe in the RVA pacing group than in the RVOT pacing group (P < 0.05), while diastolic synchrony is not significantly (NS) different between the 2 groups after pacing. There were no significant differences with respect to the mean myocardial systolic (Sm) and early diastolic velocities (Em), LV ejection fraction, LV end-diastolic and systolic volume in the 2 groups at 12 months of follow-up (all NS). CONCLUSIONS: Although RVOT pacing caused more synchronous LV contraction compared with RVA pacing, it had no benefit over RVA pacing in aspect of preventing cardiac remodeling and preserving LV systolic function after 12 months of pacing in patients with normal cardiac function. PMID- 20027662 TI - Variability of peripheral arterial tonometry in the measurement of endothelial function in healthy men. AB - Measurement of endothelial function using peripheral arterial tonometry (PAT) has been reported to be significantly correlated with coronary blood flow. Repetitive PAT measurements were performed in 22 healthy male subjects at test intervals of 1 hour (5 times within a day) and 0.5 hours (7 times within a day) to evaluate the variability of the reactive hyperemia index (RHI). A total of 10 subjects underwent additional repetitive PAT at 2 hour intervals (7 times within a day) for 3 consecutive days to evaluate the diurnal effects and day-to-day reproducibility. The RHI from each test was computed automatically based on a 15 minute recording of pulse wave amplitude changes of the fingers in response to reactive hyperemia induced by a 5 minute occlusion of the brachial artery. Intrasubject variability of RHI at different test intervals, defined as the coefficient of variation (CV) was 15.3% +/- 5.3%, 16.1%+/- 7.8%, and 22.6% +/- 3.9% for the tests at 0.5 hour, 1 hour, and 2 hour intervals, respectively. Reactive hyperemia indices measured at the same time points on each of the 3 days were not statistically significant. The interday reproducibility, presented as intraclass correlation coefficients (ICC) ranged from - 0.07 to 0.47. We conclude that repetitive PAT measurements have no carryover effect on RHI at 1 hour, and 2 hour intervals, and the RHI measured at 0.5 hour intervals is associated with a trend of increase. The interday reproducibility is relatively low and the intrasubject variability of RHI is similar to those observed in studies of flow mediated dilation using brachial artery ultrasound scanning. PMID- 20027663 TI - p53 Levels positively correlate with carotid intima-media thickness in patients with subclinical atherosclerosis. AB - BACKGROUND: The level of circulating p53 is related to inflammation in asymptomatic subjects with cardiovascular risk factors. Whether p53 is associated with the severity of atherosclerosis remains to be determined. HYPOTHESIS: This study examines the relationship of systemic p53 levels with atherosclerotic risk factors and subclinical atherosclerosis. METHODS: Circulating levels of p53 and markers of inflammation were measured in 356 subjects with cardiovascular risk factors but who were free from clinical cardiovascular disease. Subclinical atherosclerosis was evaluated by both the mean carotid intima-media thickness (IMT) and the presence of atherosclerotic plaques with the use of B-mode ultrasound in all subjects. RESULTS: p53 levels were positively correlated with age (r = 0.382, P < 0.001), intercellular adhesion molecular-1 (ICAM-1; r = 0.510, P < 0.01), vascular cell adhesion molecular-1 (VCAM-1; r = 0.497, P < 0.01), E-selectin (r = 0.337, P < 0.01), and carotid IMT (r = 0.594, P < 0.01). The association between p53 and IMT remained significant in multiple regression analysis (P < 0.01) when controlling for traditional atherosclerotic risk factors and inflammatory markers. CONCLUSION: Higher plasma p53 levels were associated with an increase in inflammatory markers, as well as increased carotid IMT. Circulating p53 may be useful in identifying subclinical atherosclerosis in subjects symptomatically free from cardiovascular disease. PMID- 20027664 TI - Plasma N-terminal protype-B natriuretic peptide and restrictive mitral flow to risk-stratify patients with stage B heart failure. AB - BACKGROUND: The prognostic role of echocardiography is well established in patients with previous or current symptoms of heart failure (HF). Less is understood about patients with stage B HF, who have structural heart disease but no symptoms at any stage. This study investigated the role of plasma N-terminal protype-B natriuretic peptide (NT-proBNP) and Doppler-echo parameters of diastolic and systolic function for risk stratification in stage B HF patients. HYPOTHESIS: We investigated whether plasma NT-proBNP and Doppler-echo parameters of diastolic and systolic function have a role in risk stratification of stage B HF patients. METHODS: Doppler echocardiography was performed in 155 outpatients with asymptomatic left ventricular (LV) dysfunction (ejection fraction [EF] < 50%) and comprised conventional diastolic variables, including pulsed-Doppler mitral E-wave deceleration time (EDT). Plasma NT-proBNP was assessed at the time of the echocardiogram. The prespecified end point was cardiac death or HF hospitalization or any new HF episode requiring a change in medication. RESULTS: Mean EF was 35% +/- 7%. Median NT-proBNP was 745 pg/mL (interquartile range [IQR], 442-1672). Patients with events at follow-up (n = 46) had higher prevalence of risk factors for developing HF. Plasma NT-proBNP, EDT, and EF were independently associated with cardiac events at multivariable analysis and receiver operating characteristic curves were used to determine the optimal prognostic cut points for each: NT-proBNP > or = 831 pg/mL, EDT < 145 msec, and EF < 37%. When used in combination, event-free survival was incrementally worse with each additional abnormality. CONCLUSION: Plasma NT-proBNP, EDT, and EF are independently useful for prognostic stratification of stage B HF patients and suggest an incremental prognostic deficit is associated with the combination of each. This study supports the complementary role of comprehensive echocardiography and natriuretic peptide assay in asymptomatic LV dysfunction. PMID- 20027665 TI - Conflict of interest. AB - Conflicts of interest are mainly financial when it comes to physician/industry relationships. Financial relationships are those in which a person benefits by receiving money in any amount in the previous 12 months. The Accreditation Council of Continuing Medical Education (ACCME) defines conflict of interest as, "when an individual has an opportunity to affect CME content about products or services of a commercial interest with which he/she has a financial relationship." PMID- 20027666 TI - Prehospital electrocardiograms (ECGs) do not improve the process of emergency department care in hospitals with higher usage of ECGs in non-ST-segment elevation myocardial infarction patients. AB - BACKGROUND: This article will describe the impact of prehospital electrocardiogram (ECG) use on emergency department (ED) processes of care for non-ST-segment elevation myocardial infarction (NSTEMI) patients and assess the characteristics associated with prehospital ECG use. METHODS: This is a retrospective, multicenter, observational analysis of NSTEMI patients captured by the National Cardiovascular Data Registry-Acute Coronary Treatment and Intervention Outcomes Network Registry-Get with the Guidelines (NCDR ACTION-GWTG) in 2007. Patient and hospital data were stratified by documentation of a prehospital ECG (pECG). Hospitals were stratified into tertiles of pECG use by higher pECG (>5.6%, n 91), lower pECG (< or = 5.6%, n = 83), or no pECG (n = 100). Statistical evaluation was done via Wilcoxon rank sum and chi(2) tests. RESULTS: There were 21 251 patients eligible for analysis. A pECG was documented in 1609 (7.6%) patients. Of 274 hospitals, 100 (36.5%) had no pECGs recorded. Median ED length of stay (LOS) was shorter at no pECG hospitals vs lower pECG hospitals (3.97 h vs 4.12 h, P < 0.05), but not higher pECG hospitals vs no pECG hospitals (3.85 h vs 3.97 h, P = not significant [NS]). A pECG was not associated with an improvement in ED performance metrics (use of aspirin, beta-blocker, any heparin) in the higher pECG hospitals vs no pECG hospitals or the lower pECG hospitals vs no pECG hospitals. CONCLUSIONS: Use of prehospital ECG in NSTEMI patients is uncommon. In contrast to its impact on reperfusion times in ST segment elevation myocardial infarction (STEMI) patients, its use does not appear to be associated with an improvement in ED processes of care at the hospital level. PMID- 20027667 TI - Retracted: Tetraoxanes: synthetic and medicinal chemistry perspective. PMID- 20027668 TI - Curcumin and its analogues: potential anticancer agents. AB - This review chronicles the exploration of the curcumin in terms of development of analogues for the anticancer activity over the last century. Curcumin is a natural phytochemical obtained from dried root and rhizome of Turmeric (Curcuma Longa). It has been shown to interfere with multiple cell signaling pathways, including apoptosis (activation of caspases and downregulation of antiapoptotic gene products), proliferation (HER-2, EGFR, and AP-1), angiogenesis (VEGF), and inflammation (NF-kappaB, TNF, IL-6, IL-1, COX-2, and 5-LOX). In the last decade it has been much explored and various synthetic analogues have been prepared and evaluated for various pharmacological activities. Most of the analogues have shown very good anticancer activity in various models and various cell lines. However, some analogues have also shown antioxidant, anti-HIV, antimutagenic, antiangiogenic, antimalarial, antitubercular, antiandrogenic, COX inhibitory activities. Few analogues have shown very potent results and may be considered as clinical candidates for the development of future anticancer agent. This review contains 728 curcumin analogues and covers the literature from 1815 to mid 2009 and 93 references are cited. PMID- 20027669 TI - Design, evaluation and structure-activity relationship studies of the AChE reactivators against organophosphorus pesticides. AB - Organophosphate pesticides (OPPs; e.g. chlorpyrifos, diazinon, paraoxon) are a wide and heterogeneous group of organophosphorus compounds. Their biological activity of inhibiting acetylcholinesterase (AChE) or butyrylcholinesterase (BChE) ranks them as life endangering agents. The necessary treatment after OPP exposure involves the use of parasympatolytics (e.g. atropine), oxime reactivators (e.g. obidoxime), and anticonvulsive drugs (e.g. diazepam). Therefore, the reactivators of AChE are essential compounds in the treatment of OPP intoxications. Commercial AChE reactivators (e.g. pralidoxime, HI-6, obidoxime, trimedoxime, methoxime) were originally developed for other members of the organophosphate family, such as nerve agents (e.g. sarin, soman, tabun, VX). Pralidoxime, HI-6, and methoxime were found to be weak reactivators of OPP inhibited AChE. Obidoxime and trimedoxime showed satisfactory reactivation against various OPPs with minor toxicity issues. During the last two decades, the treatment of OPP exposure has become more widely discussed because of growing agricultural production, industrialization, and harmful social issues (e.g. suicides). In this review is the summarized design, evaluation, and structure activity relationship studies of recently produced AChE reactivators. Since pralidoxime, over 300 oximes have been produced or tested against OPP poisoning, and several novel compounds show very promising abilities as comparable (or higher) to commercial oximes. Some of these are highlighted for their further testing of OPP exposure and, additionally, the main structure-activity relationship of AChE reactivators against OPP is discussed. PMID- 20027670 TI - The design of orally bioavailable 2, 5-diketopiperazine oxytocin antagonists: from concept to clinical candidate for premature labor. AB - A short, efficient and highly stereoselective synthesis has been developed for a series of 6-indanyl-3-alkyl-7-aryl/heterocyclic-(3R, 6R, 7R)-2, 5 diketopiperazine amides that are potent and selective oxytocin (OT) antagonists. Property-based design using an estimate of human oral absorption enabled focus to be directed to those templates with the greatest chance of delivering high bioavailability in humans. This led to the 2', 4'-difluorophenyl dimethylamide 40, a highly potent (pK(i) =9.2) and selective OT antagonist (>1,000-fold selectivity vs. the human vasopressin receptors V1a, V2, and V1b) with good oral bioavailability (>50%) in the rat and dog. Increased solubility and an improved Cyp450 profile was achieved with a range of 2'-substituted 7-(1',3'-oxazol-4'-yl) (3R,6R,7R)-2,5-diketopiperazine amides and branching at the alpha-carbon of the 3 butyl group led to a superior rat pharmacokinetic profile that resulted in the discovery of the 2'-methyl-1',3'-oxazol-4'-yl morpholine amide derivative 74 GSK221149A (Retosiban), which had the best oral exposure and bioavailability in the rat. Retosiban has sub-nanomolar affinity (K(i) =0.65 nM) for the oxytocin receptor with >1400-fold selectivity over the closely related vasopressin receptors. It has good solubility, low protein binding and has a good Cyp450 profile with no significant inhibition IC(50) >100 uM. Retosiban is >15-fold more potent at the human oxytocin receptor than atosiban (a marketed i.v, peptide OT antagonist) and it has been shown to be an effective tocolytic by i.v. and by oral administration in rats, and was selected for progression as a potential clinical candidate for preterm labor. PMID- 20027671 TI - Modern lipid-, carbohydrate-, and peptide-based delivery systems for peptide, vaccine, and gene products. AB - Research related to peptide, vaccine, and gene delivery has grown exponentially over the last decade. In this review, we discuss the development of delivery systems for peptides, gene and vaccine products. Special focus is given to different lipidation and glycosylation strategies to improve the metabolic stability and membrane permeability of therapeutics, and their targeting to specific sites. The synthetic methods for preparation of the systems are also described. PMID- 20027672 TI - Pediatric non-alcoholic fatty liver disease: preventive and therapeutic value of lifestyle intervention. AB - Nonalcoholic fatty liver disease (NAFLD), ranging from simple steatosis to nonalcoholic steatohepatitis (NASH), and eventually cirrhosis and liver failure, is seen to be increasing amongst Western children. NAFLD rates are rising in parallel with the epidemic of childhood obesity, and in particular, fatty liver evolves more easily in NASH when poor dietary habits and sedentary lifestyle are combined. In fact, its general prevalence in the child population varies between 2.6% and 10%, but increases up to 80% in obese children. Since NASH is expected to become the most common cause of pediatric chronic liver disease in the near future, there is broad interest amongst clinical researchers to move forward, both in diagnosis and treatment. Unfortunately, to date, the expensive and invasive procedure of liver biopsy is seen as the gold standard for NASH diagnosis and few noninvasive diagnostic methods can be applied successfully. Moreover, there are still no approved pharmacological interventions for NAFLD/NASH. Therefore, current management paradigms are based upon the presence of associated risk factors and aims to improve an individual's quality of life, thus reducing NAFLD-associated morbidity and mortality. Today, lifestyle intervention (diet and exercise) is the treatment of choice for NAFLD/NASH. Thus far, no study has evaluated the potential preventive effect of lifestyle intervention on children at risk of NAFLD/NASH. Future studies will be required in this area with the perspective of developing a national program to promote nutrition education and increase physical activity as means of preventing the disease in individuals at risk. Here, we outline the clinical course, pathogenesis and management of NAFLD in children, highlighting the preventive and therapeutic value of lifestyle intervention. PMID- 20027673 TI - Genetic polymorphisms in non-alcoholic fatty liver disease: clues to pathogenesis and disease progression. AB - The spectrum of non-alcoholic fatty liver disease (NAFLD) ranges from simple steatosis through steatohepatitis to advanced fibrosis and cirrhosis. Although the reason why only a minority of patients develop progressive forms of disease still remains largely unclear, recent research has identified genetic factors as a possible basis for this variation in disease presentation. Most of the studies have been focused on finding associations between advanced disease forms and selected single nucleotide polymorphisms in genes encoding various proteins involved in disease pathogenesis. Although there are many limitations regarding the study design and interpretation of published data, further carefully planned studies together with implementation of new genetic technologies will likely bring new insights into disease pathogenesis and potential benefits to the management of patients with NAFLD. PMID- 20027674 TI - Indian task force for celiac disease: current status. AB - There are limited data on celiac disease (CD) from India. The limited knowledge about CD in India might be attributed to several factors. The first meeting of the Indian Task Force for Celiac Disease was held in the Asian Institute of Gastroenterology, Hyderabad, India in December 2008. The objectives of the meeting were to focus research on prevalence of CD in the wheat-eating Northern vs the rice-eating Southern Indian population, low-budget serological assays to study the underprivileged population, to involve other medical subspecialties in CD, to suggest proper legislation regarding wheat food labeling, and to organize affordable food substitutes for patients with celiac disease. PMID- 20027675 TI - Reduced normogastric electrical activity associated with emesis: a telemetric study in ferrets. AB - AIM: To characterize the gastric myoelectric activity (GMA) and intra-abdominal pressure changes induced by emetic stimuli (apomorphine and cisplatin) in the ferret. METHODS: GMA and intra-abdominal pressure were recorded in conscious, unrestrained ferrets surgically implanted with radiotelemetry transmitters. Animals were challenged with apomorphine (0.25 mg/kg sc) and cisplatin (10 mg/kg ip), and the emetic response was quantified via direct observation and intra abdominal pressure recording for 1 h and 4 h, respectively. The GMA was analyzed by spectral analysis; the parameters used to characterize the GMA were the dominant frequency (DF) and the repartition of spectral power in the bradygastric, normogastric and tachygastric frequency ranges. RESULTS: Retches were identified on the intra-abdominal pressure trace as peaks 0.30 +/- 1.01 s in duration and 59.57 +/- 2.74 mmHg in amplitude, vomit peaks were longer (0.82 +/- 0.06 s, P < 0.01) and reached a higher pressure (87.73 +/- 8.12 mmHg, P < 0.001). The number of retches and vomits quantified via direct observation [apomorphine: 65.5 +/- 11.8 retches + vomits (R+V), cisplatin: 202.6 +/- 64.1 R+V] and intra abdominal pressure (apomorphine: 68.3 +/- 13.7 R+V, n = 8; cisplatin: 219.0 +/- 69.2 R+V, n = 8) were correlated (r = 0.97, P < 0.0001) and the timing of emesis was consistent between the 2 methods. Apomorphine induced a decrease in normogastria from 45.48% +/- 4.35% to 36.70 +/- 4.34% (n = 8, P < 0.05) but the DF of the slow waves was not changed [8.95 +/- 0.25 counts/min (cpm) vs 8.68 +/- 0.35 cpm, n = 8, P > 0.05]. Cisplatin induced a decrease in normogastria from 55.83% +/- 4.30% to 29.22% +/- 5.16% and an increase in bradygastria from 14.28% +/- 2.32% to 31.19% +/- 8.33% (n = 8, P < 0.001) but the DF (9.14 +/- 0.13 cpm) remained unchanged (P > 0.05). The GMA changes induced by cisplatin preceded the emetic response as normogastria was reduced for 1 h before the onset of emesis (57.61% +/- 5.66% to 39.91% +/- 5.74%, n = 6, P < 0.05). Peri-emesis analysis revealed that the GMA was significantly disturbed during and immediately after, but not immediately before, the emetic episodes. CONCLUSION: The induction of emesis is reliably associated with a disrupted GMA, but changes may also occur prior to and following the emetic response. PMID- 20027676 TI - MRI versus 64-row MDCT for diagnosis of hepatocellular carcinoma. AB - AIM: To compare the diagnostic capability of multidetector computed tomography (MDCT) and magnetic resonance imaging (MRI) for the detection of hepatocellular carcinoma (HCC) tumour nodules and their effect on patient management. METHODS: A total of 28 patients (25 male, 3 female, mean age 67 +/- 10.8 years) with biopsy proven HCC were investigated with 64-row MDCT (slice 3 mm native, arterial and portal-venous phase, 120 mL Iomeprol, 4 mL/s, delay by bolus trigger) and MRI (T1fs fl2d TE/TR 2.72/129 ms, T2tse TE/TR 102/4000 ms, 5-phase dynamic contrast enhanced T1fs fl3d TE/TR 1.56/4.6, Gadolinium-DTPA, slice 4 mm). Consensus reading of both modalities was used as reference. Tumour nodules were analyzed with respect to number, size, and location. RESULTS: In total, 162 tumour nodules were detected by consensus reading. MRI detected significantly more tumour nodules (159 vs 123, P < 0.001) compared to MDCT, with the best sensitivity for early arterial phase MRI. False-negative CT findings included nodules < or = 5 mm ( n = 5), < or = 10 mm ( n = 17), < or = 15 mm ( n = 12 ), < or = 20 mm ( n = 4 ), and 1 nodule > 20 mm. MRI missed 2 nodules < or = 10 mm and 1 nodule < or = 15 mm. On MRI, nodule diameters were greater than on CT (29.2 +/- 25.1 mm, range 5 140 mm vs 24.1 +/- 22.7 mm, range 4-129 mm, P < 0.005). In 2 patients, MDCT showed only unilobar tumour spread, whereas MRI revealed additional nodules in the contralateral lobe. Detection of these nodules could have changed the therapeutic strategy. CONCLUSION: Contrast-enhanced MRI is superior to 64-row MDCT for the detection of HCC nodules. Patients should be allocated to interventional or operative treatment according to a dedicated MRI-protocol. PMID- 20027677 TI - Different faces of gastroparesis. AB - AIM: To test the hypothesis that pain and affect rather than impaired emptying determine symptom severity in patients with gastroparesis. METHODS: Adult patients with documented gastroparesis were enrolled prospectively in a single center and asked to complete the Gastroparesis Cardinal Symptom Index (GCSI), Hospital Anxiety and Depression Scale (HADS), the Short Form 12 (SF-12) as quality of life index, rate pain severity and answer 10 open-ended questions. RESULTS: A total of 55 patients (44 women) participated. Idiopathic (n = 29) or diabetic (n = 11) gastroparesis and connective tissue disease (n = 8) were the most common underlying causes. Antiemetics (n = 30) and prokinetics (n = 32) were most often prescribed. Seventeen patients used opioids on a daily basis. Nausea and/or vomiting (n = 28), pain (n = 24) and bloating (n = 14) were most commonly listed as dominant symptoms. Patients subjectively attributed symptom improvement to nutritional and dietary therapy (n = 11), prokinetics (n = 11), antiemetics (n = 10) or analgesic agents (n = 3). In univariate analyses, the physical subscore of the SF-12 and HADS, but not gastric emptying delay or symptom duration significantly correlated with disease severity as measured by the GCSI. In multivariate analyses, the combination of vomiting, bloating and depression best predicted the overall impact on quality of life. CONCLUSION: The study confirms the importance of pain and affect in gastroparesis, which requires novel approaches to improve more effectively the quality of life in patients with this disorder. PMID- 20027678 TI - Association of the GNAS1 T393C polymorphism with tumor stage and survival in gastric cancer. AB - AIM: To analyze the impact of the GNAS1 T393C polymorphism on prognosis and histopathology of gastric cancer. METHODS: Genomic DNA was extracted from paraffin-embedded tissues of 122 patients with primary gastric carcinoma and from the blood of 820 healthy white individuals. Allelic discrimination was performed by quantitative real-time polymerase chain reaction. Genotyping was correlated with histopathologic parameters and with overall survival according to the Kaplan Meier approach and with multivariate analysis by multiple stepwise regression. RESULTS: Thirty-nine (32%) patients displayed a CC genotype, 57 (46.7%) a CT genotype and 26 (21.3%) a TT genotype. The frequency of the C allele (fC) in the patient group was 0.55, which was not significantly different from that of healthy blood donors. The distribution was compatible with the Hardy-Weinberg equilibrium. Analysis of clinicopathological parameters did not show any significant correlation of the T393C genotype with gender (P = 0.50), differentiation (P = 0.29), pT-category (P = 0.19), pN-category (P = 0.30), pM category (P = 0.25), R-category (P = 0.95), the classifications according to WHO (P = 0.34), Lauren (P = 0.16), Goseki (P = 1.00) and Ming (P = 0.74). Dichotomization between C+ (CC+CT) and C-genotypes (TT), however, revealed significantly more advanced tumor stages (P = 0.023) and lower survival rates (P = 0.043) for C allele carriers. CONCLUSION: The present study provides strong evidence to suggest that the GNAS1 T393C allele carrier status influences tumor progression and survival in gastric cancer with higher tumor stages and a worse outcome for C allele carriers. PMID- 20027679 TI - Elevated pro-inflammatory and lipotoxic mucosal lipids characterise irritable bowel syndrome. AB - AIM: To investigate the pathophysiology of irritable bowel syndrome (IBS) by comparing the global mucosal metabolic profiles of IBS patients with those of healthy controls. METHODS: Fifteen IBS patients fulfilling the Rome II criteria, and nine healthy volunteers were included in the study. A combined lipidomics (UPLC/MS) and metabolomics (GC x GC-TOF) approach was used to achieve global metabolic profiles of mucosal biopsies from the ascending colon. RESULTS: Overall, lipid levels were elevated in patients with IBS. The most significant upregulation was seen for pro-inflammatory lysophosphatidylcholines. Other lipid groups that were significantly upregulated in IBS patients were lipotoxic ceramides, glycosphingolipids, and di- and triacylglycerols. Among the metabolites, the cyclic ester 2(3H)-furanone was almost 14-fold upregulated in IBS patients compared to healthy subjects (P = 0.03). CONCLUSION: IBS mucosa is characterised by a distinct pro-inflammatory and lipotoxic metabolic profile. Especially, there was an increase in several lipid species such as lysophospholipids and ceramides. PMID- 20027680 TI - Wireless capsule endoscopy in detecting small-intestinal polyps in familial adenomatous polyposis. AB - AIM: To detect the prevalence of small bowel polyps by wireless capsule endoscopy (WCE) in patients with familial adenomatous polyposis (FAP). METHODS: We examined prospectively 14 patients with FAP to assess the location, size and number of small-intestinal polyps. Patients' age, sex, years of observation after surgery, type of surgery, duodenal polyps and colorectal cancer at surgery were analyzed. RESULTS: During WCE, polyps were detected in 9/14 (64.3%) patients. Duodenal adenomatous polyps were found in nine (64.3%) patients, and jejunal and ileal polyps in seven (50%) and eight (57.1%), respectively. The Spigelman stage of duodenal polyposis was associated with the presence of jejunal and ileal polyps. Identification of the ampulla of Vater was not achieved with WCE. Importantly, the findings of WCE had no immediate impact on the further clinical management of FAP patients. No procedure-related complications were observed in the patients. CONCLUSION: WCE is a promising noninvasive new method for the detection of small intestinal polyps. Further investigation is required to determine which phenotype of FAP is needed for surveillance with WCE. PMID- 20027681 TI - Sphincter of Oddi dysfunction: psychosocial distress correlates with manometric dyskinesia but not stenosis. AB - AIM: To compare postcholecystectomy patients with Sphincter of Oddi (SO) dyskinesia and those with normal SO motility to determine the psychosocial distress, gender and objective clinical correlates of dyskinesia, and contrast these findings with comparisons between SO stenosis and normal SO motility. METHODS: Within a cohort of seventy-two consecutive postcholecystectomy patients with suspected SO dysfunction, manometric assessment identified subgroups with SO dyskinesia (n = 33), SO stenosis (n = 18) and normal SO motility (n = 21). Each patient was categorized in terms of Milwaukee Type, sociodemographic status and the severity of stress-coping experiences. RESULTS: Logistic regression revealed that in combination certain psychological, sociodemographic and clinical variables significantly differentiated SO dyskinesia, but not SO stenosis, from normal SO function. Levels of psychosocial stress and of coping with this stress (i.e. anger suppressed more frequently and the use of significantly more psychological coping strategies) were highest among patients with SO dyskinesia, especially women. Higher levels of neuroticism (the tendency to stress-proneness) further increased the likelihood of SO dyskinesia. CONCLUSION: A motility disturbance related to psychosocial distress may help to explain the finding of SO dyskinesia in some postcholecystectomy patients. PMID- 20027682 TI - Balloon overtube-guided colorectal endoscopic submucosal dissection. AB - AIM: To evaluate the usefulness of a balloon overtube to assist colorectal endoscopic submucosal dissection (ESD) using a gastroscope. METHODS: The results of 45 consecutive patients who underwent colorectal ESD were analyzed in a single tertiary endoscopy center. In preoperative evaluation of access to the lesion, difficulties were experienced in the positioning and stabilization of a gastroscope in 15 patients who were thus assigned to the balloon-guided ESD group. A balloon overtube was placed with a gastroscope to provide an endoscopic channel to the lesion in cases with preoperatively identified difficulties related to accessibility. Colorectal ESD was performed following standard procedures. A submucosal fluid bleb was created with hyaluronic acid solution. A circumferential mucosal incision was made to marginate the lesion. The isolated lesion was finally excised from the deeper layers with repetitive electrosurgical dissections with needle knives. The success of colorectal ESD, procedural feasibility, and procedure-related complications were the main outcomes and measurements. RESULTS: The overall en bloc excision rate of colorectal ESD during this study at our institution was 95.6%. En bloc excision of the lesion was successfully achieved in 13 of the 15 patients (86.7%) in the balloon overtube guided colorectal ESD group, which was comparable to the results of the standard ESD group with better accessibility to the lesion (30/30, 100%, not statistically significant). CONCLUSION: Use of a balloon overtube can improve access to the lesion and facilitate scope manipulation for colorectal ESD. PMID- 20027683 TI - Stereotactic body radiotherapy for isolated paraaortic lymph node recurrence from colorectal cancer. AB - AIM: To evaluate the efficacy and complications of stereotactic body radiotherapy in localized paraaortic lymph node recurrence from colorectal cancer. METHODS: From 2003 to 2009, 7 patients with paraaortic lymph node recurrence (1-3 lesions) from colorectal cancer were treated with stereotactic body radiotherapy. Total gross tumor volumes ranged from 4 to 40 mL. The doses were escalated from 36 Gy/patient to 51 Gy/patient and were delivered in 3 fractions. RESULTS: One and 3 year overall survival rates were 100% and 71.4%, respectively, and median survival was 37 mo. Grade IV intestinal obstruction was reported in 1 of 7 patients. This patient received 48 Gy in 3 fractions with a maximum point dose to the intestine of 53 Gy and V(45Gy) = 3.6 mL. However, 6 patients received an intestinal maximum point dose of < 51 Gy and V(45Gy) of < 1 mL, and did not develop any severe complications. CONCLUSION: This pilot study suggests selected paraaortic lymph node recurrence (1-3 closed lesions) that failed to respond to chemotherapy can be potentially salvaged by stereotactic body radiotherapy. PMID- 20027684 TI - First endoscopic procedure for diagnosis and staging of mediastinal lymphadenopathy. AB - AIM: To compare a first diagnostic procedure of transbronchial needle aspiration (TBNA) with selection of endoscopic ultrasound-guided fine-needle aspiration (EUS FNA) or TBNA for mediastinal lymphadenopathy. METHODS: Sixty-eight consecutive patients with mediastinal lymphadenopathy on computed tomography (CT), who required cytopathological diagnosis, were recruited. The first 34 underwent a sequential approach in which TBNA was performed first, followed by EUS-FNA if TBNA was unrevealing. The next 34 underwent a selective approach where either TBNA or EUS-FNA was selected as the first procedure based on the CT findings. RESULTS: The diagnostic yield of TBNA as the first diagnostic procedure in the sequential approach was 62%. In the selective approach, the diagnostic yield of the first procedure was 71%. There was no significant difference in the overall diagnostic yield, but there were significantly fewer combined procedures with the selective approach. CONCLUSION: Selecting either EUS-FNA or TBNA as the first diagnostic procedure achieved a comparable diagnostic yield with significantly fewer procedures than performing TBNA first in all patients. PMID- 20027685 TI - Azithromycin-containing versus standard triple therapy for Helicobacter pylori eradication: a meta-analysis. AB - AIM: To evaluate whether adding azithromycin to first-line Helicobacter pylori (H pylori) eradication improved eradication and reduced side effects. METHODS: Eligible articles were identified by searches of electronic databases. We included all randomized trials that compared azithromycin-containing with standard triple-therapy regimens for first-line treatment of H pylori infection. Statistical analysis was performed with Review Manager 5.0.10. Sub-analyses were also performed. RESULTS: We identified 14 randomized trials (1431 patients). Pooled H pylori eradication rates were 72.01% (95% CI: 58.09%-85.93%) and 69.78% (95% CI: 66.47%-73.09%) for patients with or without azithromycin by intention-to treat analysis, and the odds ratio (OR) was 1.17 (95% CI: 0.64-2.14). The occurrence of side effects differed significantly and was 15.81% (95% CI: 12.50% 19.12%) and 25.20% (95% CI: 21.44%-28.96%) for treatment with or without azithromycin, respectively, and the summary OR was 0.58 (95% CI: 0.41-0.82). Furthermore, the azithromycin-containing group had a lower occurrence of diarrhea, nausea and taste disturbance. CONCLUSION: Our review suggests that azithromycin-containing triple-therapy regimens could be equally effective in eradication of H pylori compared with standard first-line triple-therapy regimens. PMID- 20027686 TI - Performance value of high risk factors in colorectal cancer screening in China. AB - AIM: To analyze the performance value of high risk factors in population-based colorectal cancer (CRC) screening in China. METHODS: We compared the performance value of the immunochemical fecal occult blood test (iFOBT) and other high risk factors questionnaire in a population sample of 13 214 community residents who completed both the iFOBT and questionnaire investigation. Patients with either a positive iFOBT and/or questionnaire were regarded as a high risk population and those eligible were asked to undergo colonoscopy. RESULTS: The iFOBT had the highest positive predictive value and negative predictive value in screening for advanced neoplasia. The iFOBT had the highest sensitivity, lowest number of extra false positive results associated with the detection of one extra abnormality for screening advanced neoplasias and adenomas. A history of chronic cholecystitis or cholecystectomy, chronic appendicitis or appendectomy, and chronic diarrhea also had a higher sensitivity than a history of adenomatous polyps in screening for advanced neoplasias and adenomas. The sensitivity of a history of chronic cholecystitis or cholecystectomy was highest among the 10 high risk factors in screening for non-adenomatous polyps. A history of chronic appendicitis or appendectomy, chronic constipation, chronic diarrhea, mucous and bloody stool, CRC in first degree relatives, malignant tumor and a positive iFOBT also had higher sensitivities than a history of adenomas polyps in screening for non adenomatous polyps. Except for a history of malignant tumor in screening for non adenomatous polyps, the gain in sensitivity was associated with an increase in extra false positive results associated with the detection of one extra abnormality. CONCLUSION: The iFOBT may be the best marker for screening for advanced neoplasias and adenomas. Some unique high risk factors may play an important role in CRC screening in China. PMID- 20027687 TI - High expression of osteoglycin decreases gelatinase activity of murine hepatocarcinoma Hca-F cells. AB - AIM: To investigate the possible correlation between osteoglycin expression and gelatinase activity of mouse hepatocarcinoma Hca-F cells. METHODS: A eukaryotic expression plasmid pIRESpuro3 osteoglycin(+) was constructed and transfected into Hca-F cells to investigate the possible correlation between osteoglycin expression and gelatinase activity of Hca-F cells cultured with extract of lymph node, liver, spleen or in DMEM medium. The activity of gelatinases was examined through zymographic analysis. RESULTS: High expression of osteoglycin attenuated the gelatinase activity of Hca-F cells cultured with extract of lymph node, and at the same time, decreased the metastatic potential of Hca-F cells to peripheral lymph nodes in vivo. CONCLUSION: High expression of osteoglycin decreases the gelatinase activity of Hca-F cells cultured with extract of lymph node; regulation of gelatinase activity might be one of mechanisms that osteoglycin contributes to lymphatic metastasis suppression. PMID- 20027688 TI - Meckel's diverticulum manifested by a subcutaneous abscess. AB - This case report describes an extremely rare complication of a Meckel's diverticulum: enterocutaneous fistula of the diverticulum. The presence of Meckel's diverticulum is a well known entity, but subcutaneous perforation of the diverticulum is very rare. Here we report the case of a patient with the complaint of a right lower quadrant abscess, preoperatively diagnosed as enterocutaneous fistula, which was determined intraoperatively to be a fistula resulting from Meckel's diverticulum. PMID- 20027689 TI - Carcinoma of the papilla of Vater following treatment of pancreaticobiliary maljunction. AB - Pancreaticobiliary maljunction (PBM) is frequently associated with biliary cancer due to reflux of pancreatic enzymes into the choledochus, and even after surgery to correct the PBM such patients still have a risk of residual bile duct cancer. Here, we report the case of a 59-year-old female with carcinoma of the papilla of Vater which developed 2.5 years after choledochoduodenostomy for PBM. During the postoperative follow-up period, computed tomography obtained 2 years after the first operation demonstrated a tumor in the distal end of the choledochus, although she did not have jaundice and laboratory tests showed no abnormalities caused by the previous operation. As a result, carcinoma of the papilla of Vater was diagnosed at an early stage, followed by surgical cure. For early detection of periampullary cancer in patients undergoing surgery for PBM, careful long-term follow-up is needed. PMID- 20027690 TI - A case of hypereosinophilic syndrome presenting with intractable gastric ulcers. AB - We report a rare case of hypereosinophilic syndrome (HES) presenting with intractable gastric ulcers. A 71-year-old man was admitted with epigastric pain. Initial endoscopic findings revealed multiple, active gastric ulcers in the gastric antrum. He underwent Helicobacter pylori (H pylori) eradication therapy followed by proton pump inhibitor (PPI) therapy. However, follow-up endoscopy at 4, 6, 10 and 14 mo revealed persistent multiple gastric ulcers without significant improvement. The proportion of his eosinophil count increased to 43% (total count: 7903/mm(3)). Abdominal-pelvic and chest computed tomography scans showed multiple small nodules in the liver and both lungs. The endoscopic biopsy specimen taken from the gastric antrum revealed prominent eosinophilic infiltration, and the liver biopsy specimen also showed eosinophilic infiltration in the portal tract and sinusoid. A bone marrow biopsy disclosed eosinophilic hyperplasia as well as increased cellularity of 70%. The patient was finally diagnosed with HES involving the stomach, liver, lung, and bone marrow. When gastric ulcers do not improve despite H pylori eradication and prolonged PPI therapy, infiltrative gastric disorders such as HES should be considered. PMID- 20027691 TI - A special growth manner of intrahepatic biliary cystadenoma. AB - We report a case of a 56-year-old woman with intrahepatic biliary cystadenoma (IBC) accompanying a tumor embolus in the extrahepatic bile duct, who was admitted to our department on October 13, 2008. Imaging showed an asymmetry dilation of the biliary tree, different bile signals in the biliary tree, a multiloculated lesion and an extrahepatic bile duct lesion with internal septation. A regular left hemihepatectomy en bloc was performed with resection of the entire tumor, during which a tumor embolus protruding into the extrahepatic bile duct and originating from biliary duct of segment 4 was revealed. Microscopically, the multiloculated tumor was confirmed to be a biliary cystadenoma with an epithelial lining composed of biliary-type cuboidal cells and surrounded by an ovarian-like stroma. An aggressive en bloc resection was recommended for the multiloculated lesion. Imaging workup, clinicians and surgeons need to be aware of this different presentation. PMID- 20027692 TI - Comments on the editorial by Riggio & Ageloni on the ascitic fluid analysis. AB - Angeloni et al published a landmark study on the use of Coulter counters in spontaneous bacterial peritonitis (SBP) diagnosis. Riggio and Angeloni have recently published an editorial on the ascitic fluid analysis in diagnosis and monitoring of SBP. Herein, some points of interest are discussed. PMID- 20027693 TI - Relationship between abdominal trauma or surgery and mesenteric panniculitis. AB - Mesenteric panniculitis is a rare disease characterized by chronic non-specific inflammation of mesenteric fat tissue. Several etiologic and/or associated factors have been reported in the literature so far. Although trauma or surgery is one of the potential etiologic factors for mesenteric panniculitis, to the best of our knowledge, no strong correlation has been shown in the literature until now. PMID- 20027694 TI - I have atrial fibrillation and take warfarin, digoxin and metoprolol. I still get an occasional mild attack, however. Is this normal? PMID- 20027695 TI - [On the occasion of the 90th birthday anniversary of E. V. Gembitskii, chief therapist of the Armed Forces]. PMID- 20027696 TI - John Anderson's development of (situational) realism and its bearing on psychology today. AB - In 1927, the Scottish philosopher John Anderson arrived in Australia to take up the chair of Philosophy at the University of Sydney. By the late 1930s, the "macrostructure" of his realist system was in place. It includes a theory of process and a substantial metaphysics, one that opposes positivism, linguistic philosophy and all forms of idealism. However, beyond Australia it remains largely unknown, despite its bearing on a number of current issues in psychology and the social sciences generally. This article outlines Anderson's transition from Hegelian idealism to realism, describes aspects of his ontology and epistemology, compares some of Anderson's ideas with Dewey's pragmatism and explains their relevance to present-day psychology. PMID- 20027697 TI - Movement as utopia. AB - Opposition to utopianism on ontological and political grounds has seemingly relegated it to a potentially dangerous form of antiquated idealism. This conclusion is based on a restrictive view of utopia as excessively ordered panoptic discursive constructions. This overlooks the fact that, from its inception, movement has been central to the utopian tradition. The power of utopianism indeed resides in its ability to instantiate the tension between movement and place that has marked social transformations in the modern era. This tension continues in contemporary discussions of movement-based social processes, particularly international migration and related identity formations, such as open borders transnationalism and cosmopolitanism. Understood as such, utopia remains an ongoing and powerful, albeit problematic instrument of social and political imagination. PMID- 20027698 TI - Masculinity on trial: penises, hermaphrodites and the uncertain male body in early modern France. PMID- 20027699 TI - Pain and degradation in Georgian London: life in the Marshalsea prison. PMID- 20027700 TI - Courting the pink pound: "Men Only" and the queer consumer, 1935-39. PMID- 20027701 TI - India's fossil biota: current perspectives and emerging approaches. PMID- 20027702 TI - ["Model girls": the education of women in Croatia during the second half of the 19th century]. PMID- 20027704 TI - Socially not so dead! Slave identities in Bourbon Nueva Granada. PMID- 20027705 TI - [The body of history: the virgin orphan in El Carnero]. PMID- 20027706 TI - Note from the editor: "Ultrasound soft markers of chromosomal abnormalities; an ethical dilemma for obstetricians". PMID- 20027707 TI - Problems of Parliamentary accountability in Jamaica: consequences for health administration. AB - The effective parliamentary democracy implements the policies, programmes, procedures and processes that encourage optimal constituency service, effective departmental performance, thoroughly considered and well-formulated government policies, public responsiveness and accountability. This paper uses a case study of health administration to highlight some of the problems presented in seeking to establish parliamentary accountability in Jamaica. It argues that the ineffectiveness and lack of accountability in and of Parliament have flourished in a context of poor economic growth and deep political and social divides. Accountability requires more than ad-hoc institutional interventions; it depends on social and political change. PMID- 20027708 TI - "An epidemic of runaway wives": discourses by Dani men on sex and marriage in highlands Irian Jaya, Indonesia. PMID- 20027709 TI - Pneumococcal vaccination: effective in children under the age of 2 years. AB - In France, since the introduction of pneumococcal vaccination for children under 2 years of age, the incidence of pneumococcal meningitis and bacteraemia has declined in this age group. PMID- 20027710 TI - Complex regional pain syndrome type 1. Some treatments assessed versus placebo, limited efficacy. AB - (1) Complex regional pain syndrome type 1 generally occurs after trauma and usually affects a limb; (2) How is complex regional pain syndrome type 1 diagnosed? What is its natural course? How safe and effective are available treatments? To answer these questions, we reviewed the literature using the standard Prescrire methodology; (3) Diagnosis is mainly based on clinical features, including pain disproportionate to the initial trauma, associated with cutaneous vasomotor, trophic and sweating disorders; (4) Some clinical signs call for additional examinations to help rule out another vascular, neurological, infectious or rheumatic disorder. Radiological evidence of bone demineralisation supports the diagnosis, but radiography, magnetic resonance imaging (MRI) and scintigraphy generally contribute little to the diagnosis of complex regional pain syndrome; (5) Some patients recover spontaneously after a few weeks, while others develop chronic pain or even severe disability after a period of years; (6) The results of small placebo-controlled trials suggest that corticosteroids are effective during the initial phase of this syndrome; (7) A very high oral dose of alendronic acid provided sustained pain relief in a randomised trial. Other studies suggest that bisphosphonates have some impact. The adverse effects of alendronic acid given at such high doses are poorly known; (8) Calcitonin, antiepileptics, antidepressants and opiates have no proven efficacy; (9) Transcutaneous neurostimulation is rapidly effective and safe, but its efficacy also diminishes rapidly. Therefore, the sessions have to take place at increasingly shorter intervals. (10) Spinal neurostimulation with implanted electrodes has been assessed in a comparative trial in 54 patients. Some efficacy was observed, but one-third of patients had complications requiring further surgery; (11) Various substances have been given intravenously with the goal of achieving regional anaesthesia, but none was found to have any tangible efficacy; (12) Sympathectomy has a negative risk-benefit balance; (13) Non-drug approaches such as physiotherapy, relaxation and biofeedback seem to help some patients, with little risk of adverse effects. These methods have not been comparatively evaluated; (14) In practice, there is no truly effective treatment for complex regional pain syndrome. The few beneficial treatments have not been directly compared with one another. The advantages and disadvantages of the various treatment options must be discussed with each patient. PMID- 20027711 TI - [Robbery and larceny in Mexico City in the late 18th century]. PMID- 20027712 TI - [Spaces and practices of sociability in the 18th century: social gatherings, refreshments, and cafes of Barcelona]. PMID- 20027713 TI - Bell's palsy: corticosteroids effective. AB - Two randomised double-blind trials in about 1400 patients showed that for every 7 to 8 patients treated with prednisolone, one additional patient will recover normal motor function after 9 to 12 months. PMID- 20027714 TI - Antiplatelet drugs for patients at high cardiovascular risk. Aspirin generally remains the best choice. PMID- 20027715 TI - Pre-eclampsia: aspirin beneficial. AB - Two meta-analyses suggest that low-dose aspirin taken daily during pregnancy reduces the incidence of pre-eclampsia in women at risk and reduces mortality among the infants born to mothers at high risk of pre-eclampsia. PMID- 20027716 TI - [Revolutionary acculturation: French struggles for a new sociability, 1789-1799]. PMID- 20027717 TI - Childhood psoriasis: often favorable outcome. AB - (1) Plaque psoriasis is the most common form of psoriasis in children. Topical agents should be tried first, especially well-tolerated products such as emollients. Topical corticosteroids are sometimes useful during exacerbations but, given adverse effects, they should only be used for short periods; (2) UVB phototherapy is an option for extensive psoriasis refractory to local treatments, but it carries a long-term risk of skin cancer. Immunosuppressants have not been well assessed in this setting, but methotrexate has been better evaluated than the others. PMID- 20027718 TI - Gene therapy: concepts and methods. Few applications so far. AB - (1) Gene therapy consists of inserting foreign genetic material into a patient's cells to correct a health disorder; (2) Only a fraction of gene therapy trials involve hereditary single-gene disorders. Most trials focus on acquired disorders such as cancer and cardiovascular disease; (3) The therapeutic gene can be delivered directly into the target organ or the general circulation, or alternatively into cells that are first isolated from the patient and then reinjected; (4) One of the main problems with gene therapy is finding an appropriate vector (viral or non-viral) that is both effective and safe. Most currently available vectors confer only weak and transient therapeutic gene expression. In addition, many carry major risks such as immunological disorders and oncogenicity; (5) In early 2009, only one therapeutic gene product is commercially available, in China. PMID- 20027719 TI - [Francisco Monzon and "the good sleep": the theological interpretation of dreams in Spain in the 16th century]. PMID- 20027720 TI - The role of genes in disease: beware of simplistic interpretations! AB - (1) Most diseases with a strong genetic component and a Mendelian pattern of inheritance are rare or very rare disorders; (2) The most common human diseases result from complex interactions between genetic and environmental factors. They are multifactorial; (3) The simple presence of genes that are associated with a disease is not generally sufficient to determine with certainty that the signs and symptoms that characterise the disease will develop in that individual; (4) The association of a disease with a gene does not mean that the gene causes the disease. PMID- 20027721 TI - Who controls this private space? The offense and defense of the hoop in early eighteenth-century France and England. PMID- 20027722 TI - Fighting over the reinterpretation of the Mongolian woman in Mongolia's post socialist identity construction discourse. PMID- 20027723 TI - Women's International War Crimes Tribunal on Japan's military sexual slavery: memory, identity, and society. PMID- 20027726 TI - Developing the medical model. PMID- 20027727 TI - Filing claims for glaucoma care. PMID- 20027728 TI - Little need for "hand holding" after the purchase of a practice. PMID- 20027729 TI - Consensus of the French Society of Gerontology and Geriatrics and the French Society of Cardiology for the management of coronary artery disease in older adults. AB - Coronary heart disease is a common and serious condition in patients aged over 80 years. The presenting clinical symptoms are all the more atypical and the prognosis poorer when it occurs in patients with multiple comorbid diseases. The presence of comorbidities dictates the need for a standardized geriatric assessment to screen for the existence of underlying frailty. The available scientific data were obtained during studies that included few subjects aged over 80 years. These recommendations are therefore mainly extrapolated from results obtained in younger populations. The pharmacological management and revascularization strategy for coronary heart disease in octogenarians is basically the same as in younger subjects. Epidemiological studies all concur that available therapies are underutilized despite the fact that this population has a high cardiovascular risk. Specific precautions for use must be respected because of the comorbidities and age-related changes in pharmacokinetics or pharmacodynamics. Generally, the therapeutic strategy in coronary heart disease is based not on the patient's real age, but rather on an individual analysis taking into account the severity of the coronary disease, comorbidities, the risk of drug misadventures, patient life expectancy and quality of life. PMID- 20027730 TI - [Development of abdominal pain and abdominal bloating following hemodialysis- probable diagnosis based on plain abdominal CT]. PMID- 20027731 TI - Reducing child poverty in Britain: an assessment of government policy, 1997-2001. PMID- 20027732 TI - Housing subsidies and work incentives in Great Britain. PMID- 20027733 TI - The demand for private medical insurance in the UK: a cohort analysis. PMID- 20027734 TI - Modernism, male intimacy, and the Great War. PMID- 20027737 TI - Medical inspection of State schoolchildren in Australia, c. 1905-14. AB - From about 1905 some medical practitioners pressed State governments to back official physical examinations of State school children. Their campaign embodied a broader project to identify and reduce incapacities undermining public hygiene and national efficiency. Some of the activists became inspectors in Tasmania, Victoria, South Australia and Queensland. Evidence about their relations with children, teachers, parents, communities and local GPs seems to be meagre but worth pursuing. The campaign's immediate and longterm effects on child health apparently disappointed its proponents 'hopes, but still might have helped many children and benefited national fitness. Later repercussions of boys being classed as 'fit' could prove deadly in the Great War. PMID- 20027738 TI - In the 'Grey Battalion': Launceston General Hospital nurses on active service in World War I. AB - Nearly fifty nurses from the Launceston General Hospital served with the Australian Army Nursing Service or the British nursing service during World War I. These nurses served in countries that included Egypt, France, India, Greece, Italy and England. They worked in various roles including as a surgical team nurse close to the front working under fire; nursing on hospital ships carrying the wounded; or managing wards overrun with patients on ships at Gallipoli in 1915, whilst dealing with a lack of hospital necessities. This case study, of one group of nurses, shows that their experiences reflected the skills and roles needed to be a military nurse of the time, significantly different to the skills required to nurse in Australia. PMID- 20027739 TI - Fantome Island lock hospital and aboriginal venereal disease sufferers 1928-45. AB - In this article the Queensland government's response to suspected and confirmed cases of venereal disease amongst the state's Aboriginal population is examined through the micro history of Fantome Island lock hospital, which operated between 1928 and 1945. This history offers an interesting case study into the complexities of medical and racial segregation in twentieth century Queensland. While other scholars have positioned Fantome Island lock hospital as a justifiable attempt to control syphilis and gonorrhoea infections amongst the Queensland Aboriginal population, I propose a different interpretation and argue that white perceptions of Aboriginal sexuality and health contributed to government depictions of an Aboriginal venereal disease 'epidemic.' I demonstrate that disease diagnosis was still highly problematic prior to World War II and was differentially applied across different sub-populations. PMID- 20027740 TI - Determining the meaning behind historical disease terminology through an examination of patterns of terminology used in the mortality statistics of Victoria, 1853-1900. AB - Presented are four methods which may be used to determine the meaning behind past terminology usage, demonstrated through the utilisation of Victorian Registrar General Reports from 1853 to 1900. These methods are: (1) looking for direct changes in terminology usages; (2) use of a control disease; (3) comparing historical terms whose use mirrors each other; and (4) using risk factor statistics. Through these methods, it is concluded that in the second half of the nineteenth century the term 'hydrocephalus' referred to tubercular meningitis, 'intemperance' to alcohol poisoning, 'quinsy' and 'laryngitis' indicated a diphtheria infection, 'puerperal fever' referred to group A streptococci infection, 'typhoid/typhus' described typhoid more than typhus, 'teething' often indicated infantile diarrhoea and 'tumour' often a tapeworm infection. PMID- 20027741 TI - Breasts for hire and shared breastfeeding: wet nursing and cross feeding in Australia, 1900-2000. AB - Wet nursing and cross-nursing both involve the breastfeeding of a baby by someone who is not the baby's mother. They differ in that wet nurses were usually employees in paid situations and the breastfeeding was not reciprocated, whereas cross-nursing was between peers and was usually unpaid and could be reciprocal. In Australia at the turn of the twentieth century wet nurses were employed in private homes or institutions. By the 1920s, finding a wet nurse had become extremely difficult, but informal cross-feeding existed long after this. This study examines both practices in twentieth century Australia, 1900-2000, and includes a discussion of the decline of human milk banks, another means of sharing mothers' milk. PMID- 20027742 TI - The value of an infant: the rise of paediatrics in Australia, 1880-1910. AB - Between 1880 and 1910 paediatrics in Australia developed not merely as a response to Enlightenment philosophical understandings of the child as precious and special, but as part of a wider demand for reproduction and population. A brief sketch of the international context will situate the specific Australian conditions, which include education, professionalisation and the emerging concept of infant mortality. A level of general specialisation within medicine was necessary for the development of paediatrics, in addition to a general and new interest in child health, which was a response to the social, political and economic needs of the emerging nation. PMID- 20027743 TI - Fleas, ruby and the power of chiropractic. AB - This essay details my experience with a woman who believed her profound incapacity was due to her failure to respect what she termed the power of chiropractic. Whilst the clinical setting may be a fertile locale for the analysis of power the essay seeks not to allocate supplementary relations or uncover unspoken truths by privileging none but a remarkable woman's interpretation of that power. PMID- 20027744 TI - Dr Thomas Beddoes: chemistry, medicine, and the perils of democracy. AB - Beddoes lectured on chemistry at Oxford in the years that included the French Revolution, the Terror, and the outbreak of war with France, as well as the success in France of the chemical revolution. The very public dispute between Edmund Burke and Joseph Priestley meant that the latter's study of different kinds of air was politically tainted. Beddoes's democratic beliefs and his support for the new chemistry of Lavoisier meant that as chemist and physician he had to deal with complaints that he was potentially seditious and pro-French. His medical theories, allied to pneumatic chemistry and building on the work of Priestley, were accordingly suspect. In spite of that, he became the physician and friend to several members of the Lunar Society of Birmingham and to members of their family, and they in return became his patrons. His collaboration with James Watt was crucial for his development of pneumatic medicine. The full extent of Lunar patronage, and especially that of James Keir and Thomas Wedgwood, has hitherto not been recognized, but it was the concealed scale of that patronage that made possible the execution of Beddoes's ambitious programme of treatment and research. PMID- 20027745 TI - His majesty's subjects: from laboratory to human experiment in pneumatic chemistry. AB - Experiments in pneumatic chemistry paved the way for medical innovation in the last quarter of the eighteenth century. Thomas Beddoes and James Watt were instrumental in the spread of the use of new gas chemistry in pneumatic therapy, but they were far from alone. There was no shortage of experimental subjects, as the practice was quickly taken up by medics throughout Britain. PMID- 20027746 TI - Political gout: dissolute patients, deceitful physicians, and other blue devils. AB - This essay seeks to assess the renegade Thomas Beddoes through the filter of the gout diagnosis in his time. It stretches out to cover his whole life and emphasizes the need for a broad comparative historical and biographical approach. Gout is shown to have functioned then as more than a malady; it was also part of a social code embedding class, rank, affiliation, standing and political position. PMID- 20027747 TI - The atmosphere of heaven: the 1799 nitrous oxide researches reconsidered. AB - Thomas Beddoes's and Humphry Davy's accounts of the nitrous oxide experiments carried out at the Pneumatic Institution in 1799 include extravagant descriptions of its mind-altering effects. Many people, both at the time and subsequently, have considered these descriptions to be the product not of the gas but of its subjects' overheated imaginations. To what extent were these effects 'all in the mind' of the experimenters? Modern understandings of nitrous oxide throw new light on this question; but it was also considered, and resolved in different ways, by Beddoes and Davy themselves. PMID- 20027748 TI - Thomas Beddoes and the German psychological tradition. AB - This paper considers Thomas Beddoes's role in disseminating German psychological ideas in Britain. It describes the German tradition as inaugurated by Karl Philipp Moritz (1756-93) and considers the chief differences between this tradition and the English one stemming from David Hartley. It is suggested that Beddoes found strong support for his convictions about human interiority in writings by Moritz and his followers. In particular, these enabled him to think about sanity and madness as being continuous with one another (rather than as one another's negation); they helped him locate the signs of madness in ordinary childhood behaviours; they reinforced his suspicions that many so-called nervous disorders were psychically caused; and they supplied him with a conception of unconscious passion. The paper concludes by considering Beddoes's appeal to Shakespeare's plays as a source of clinical knowledge about the nature of insanity, and argues that Beddoes has been overlooked as a crucial source for nineteenth-century psychiatrist-bardologists such as J. Conolly, J. C. Bucknill and H. Maudsley. PMID- 20027749 TI - A welfare trap? The duration and dynamics of social assistance use among lone mothers in Canada. AB - In the late 1990s, the Canadian social assistance policy changed to focus on reducing "dependency" or long-term receipt. Using data from the Survey of Labour and Income Dynamics (1996-2001) and event history models, we investigate the duration of social assistance receipt for lone mothers and other household heads. We find that lone mothers' education and labor force experience were less important predictors for their duration on social assistance than their previous marital history. Although receipt was generally short term, we find evidence of negative duration dependence, or a "welfare trap," after controlling for unobserved heterogeneity. This indicates one way in which a welfare receipt is not only the result of particular life course trajectories but also shapes lives. PMID- 20027750 TI - Privatizing responsibility: public sector reform under neoliberal government. AB - In light of public sector reforms in Canada and elsewhere, this paper focuses on the shift of emphasis from social to private responsibilities and raises new questions about the forces of private enterprise and market-based partnerships. Under neoliberal governmental agendas, privatizing responsibility links to three main developments: the reconsideration of the relations of public and private; the mobilization of responsible citizenship; and the formation of a cultural mentality of rule that works alongside these developments. The research for this article is based on extensive analysis of policy documents and public sector reform initiatives, as well as interviews with Canadian federal public service employees. PMID- 20027751 TI - "Choice" in filial care work: moving beyond a dichotomy. AB - In this paper, we discuss the concept of"choice" in filial care work. A dichotomy between choice and obligation is constructed in theoretical and empirical literature on filial care work. However, this dichotomy does not adequately reflect family caregiving experiences; for instance, choice and obligation are not mutually exclusive motivations. Rather, choice and obligation are "relational" and "contextual" and manifest in the interaction between macro- and micro-contexts. The choice-obligation dichotomy is also ideological. At the moral political level, it promotes the downloading of responsibility by governments to family, obscures the complexity of filial relationships, and may even promote psychological ambivalence. PMID- 20027752 TI - Policy change and the politics of ideas: the emergence of the Canada/Quebec Pension Plans. AB - Stressing the direct impact of ideas on policy change, this article explores the adoption of the Canada and Quebec Pension Plans (C/QPP) in 1965 by addressing two closely related questions: in the mid-1960s: why did the federal government decide to create an earnings-related public pension system on top of the existing Old Age Security program? Second, why did that new system feature a replacement rate higher than initially proposed as well as a separate scheme for the province of Quebec? In order to answer these two questions, the article analyzes the debates leading to the enactment of the C/QPP. PMID- 20027753 TI - Art and the history of medicine. The dwarf pointed by Mantegna of Montua and the Morgante of Florence. AB - This short article concerns two images which relate art and the history of medicine. The first is a fresco by Andrea Mantegna, from the period 1465-1474, when he was at the Gonzaga court of Ludovic III. This painting includes a female dwarf. Modern ideas on dwarfism are briefly discussed and another dwarf, Morgante, from the court of the Medicis in Florence, is described and illustrated. PMID- 20027754 TI - [A tribute to prominent African American dentists]. AB - If today, American society has undeniably changed, this has been made possible by the selflessness, courage and perseverance of previous generations. Among them are the black American dentists, who met constant opposition. This paper pays homage to their memory. PMID- 20027755 TI - The contribution of Ukrainian physicians to world medicine. AB - This paper gives information about fifty physicians, from the Middle Ages to the present, who have enriched world medicine. It describes the contribution made both by Ukrainian physicians and those of other nationalities, who worked in Ukrainian lands occupied by Russia, Poland, Austria-Hungary and Romania. The paper notes the contribution that these physicians have made to modern medical science, paying special attention to their achievements and published work. PMID- 20027756 TI - Michelangelo: anatomy and its implication in his art. AB - Michelangelo's major interest was the Life of the Soul as expressed in the beautiful structure and movement of the human body, which he often called the "mortal veil" of the divine intentions. This study ascertains Michelangelo's interest in and acquisition of the knowledge of human anatomy, the use of small anatomical models to crystallize his concepts into reality and the application of anatomy to his art. Relatively little is known of this interaction between anatomy and art in Michelangelo's life and work. PMID- 20027757 TI - Hitler's bible: an analysis of the relationship between American and German eugenics in pre-war Nazi Germany. AB - Throughout the last century the wellbeing of those with disability has been threatened by the idea of eugenics. The most notable and extreme example of this could be considered to have been carried out during World WarTwo, within Nazi eugenic programmes. These resulted in the sterilisation and killing of hundreds of thousands of disabled people. Through research of a wide range of sources it has been established that much of the inspiration and encouragement for this rapidly progressing movement in Germany initially came from America, most notably from California. American eugenicists expressed interest, and at times jealousy, at the speed of the progression in German eugenics. German Sterilisation laws were drafted following careful study of American experiments and research, while financial support from a number of American individuals encouraged further German research. Correspondence between influential leaders, including Hitler, Grant and Whitney, Verschuer and Popenoe, on both sides also added to the developing relationship. In conclusion, although there are a number of vital differences between the progress of the eugenics programme in America and in pre-war Nazi Germany, and eugenics in America never produced the massive genocide that occurred in Germany, it is clear that the research, encouragement and enthusiasm from America had a profound influence on the rapidly growing Nazi eugenics movement. PMID- 20027758 TI - A little known episode in the life of the future revolutionary Fabre d'Eglantine: his involvement in mephitism. AB - On one hand we have Philippe-Francois-Nazaire Fabre, known as Fabre d'Eglantine, an undistinguished playwright, who, in September 1792, during the French Revolution, was elected a member of the Convention nationale and voted for the execution of Louis XVI in January 1793. On the other hand we have Jean-Antoine Michel-Dieudonne Janin de Combe-Blanche, one of the most famous ophthalmologists of the eighteenth century, raised to the peerage in 1787, physician to crowned heads and dignitaries of the Church. These two men were diametrically opposed, but were brought together by chance, shortly before the outbreak of the French Revolution, by a pamphlet on mephitism. PMID- 20027759 TI - Monstrous births and medical networks: debates over forensic evidence, generation theory, and obstetrical authority in France, ca. 1780-1815. AB - In France between 1780 and 1815, doctors opened a broad correspondence with medical faculties and public officials about foetal anomalies ("monstrosities"). Institutional and legal reforms forced doctors to encounter monstrous births with greater frequency, and they responded by developing new ideas about heredity and embryology to explain malformations to public officials. Though doctors achieved consensus on pathogenesis, they struggled to apply these ideas in forensic cases, especially with doubtful sex. Medical networks simultaneously allowed doctors to explore obstetrical techniques, as licensing regulations forced practitioners into emotional encounters with child anomalies. Doctors thus developed a new ethics for treating monstrosities, viewing them as pathological specimens, forensic objects, and obstetrical tragedies. PMID- 20027760 TI - The mine and the furnace: Francis Bacon, Thomas Russell, and early Stuart mining culture. AB - Notwithstanding Francis Bacon's praise for the philosophical role of the mechanical arts, historians have often downplayed Bacon's connections with actual artisans and entrepreneurs. Addressing the specific context of mining culture, this study proposes a rather different picture. The analysis of a famous mining metaphor in The Advancement of Learning shows us how Bacon's project of reform of knowledge could find an apt correspondence in civic and entrepreneurial values of his time. Also, Bacon had interesting and so far unexplored links with the early modern English mining enterprises, like the Company of Mineral and Battery Works, ofwhich he was a shareholder. Moreover, Bacon's notes in a private notebook, Commentarius Solutus, and records of patents of invention, allow us to start grasping Bacon's connections with the metallurgist and entrepreneur Thomas Russell. Lastly, this paper argues that, to fully understand Bacon's links with the world of Stuart technicians and entrepreneurs, it is necessary to consider a different and insufficiently studied aspect of Bacon's interests, namely his work as patents referee while a Commissioner of Suits. PMID- 20027761 TI - Early modern green sickness and pre-Freudian hysteria. AB - In early modern medicine, both green sickness (or chlorosis) and hysteria were understood to be gendered diseases, diseases of women. Green sickness, a disease of young women, was considered so serious that John Graunt, the father of English statistics, thought that in his time dozens of women died of it in London every year. One of the symptoms of hysteria was that women fell unconscious. The force of etymology and medical tradition was so strong that in one instance the gender of the patient seems to have been changed by the recorder to make the case fit medical theory. PMID- 20027763 TI - Saving grace. Health savings accounts are getting more popular--and easier for your practice to accept. PMID- 20027762 TI - Go team! How an integrated care group turns the tide against cardiometabolic disorders. PMID- 20027764 TI - At the first hint of litigation, contact your insurer. PMID- 20027765 TI - A welcome gift. A former patient provides the antidote to burnout. PMID- 20027766 TI - [Differentiation and synthesis. Forms of reception of acoustical research in the musical literature of the nineteenth century]. AB - In the nineteenth century, both musical scholars and natural scientists discussed the relevance of acoustical research for the theory and practice of music. Whereas some musical theorists and acousticians plead together for an acoustical foundation of musical theory, other scholars questioned the significance of physical and physiological knowledge for a deeper understanding of music. Based on an analysis of musical journals, popular scientific writings, theoretical treatises and musical dictionaries this article demonstrates how musical scholars and natural scientists argued about the question which discipline should have the final say about musical concepts and terminologies. To merge both heterogeneous spheres--music and acoustics--or to carefully distinguish between them--these two positions shaped the dispute over the relationship between music and natural sciences in the nineteenth century. PMID- 20027767 TI - [Tone psychology and music research as catalysts of experimental-scientific practice and methodology in the circle of Carl Stumpf]. AB - The study of acoustics, harmonics and of music has been providing scientific models since Greek Antiquity. Since the early modern ages, two separate cultures began to emerge out of the study of music: a technical acoustics and an aesthetically and philosophically inspired musical criticism. In the writings of Johann Friedrich Herbart (1811) a scientific approach to musical aesthetics and to music perception is taking shape that reinstalls the listening process as a highly complex and logical phenomenon. By opening music for a scientific psychological investigation, Herbart pioneered the physiologically and acoustically grounded seminal work by Hermann von Helmholtz On the sensations of tone (1863) which the author considered a prerequisite for musical aesthetics and music theory. Helmholtz in turn inspired the philosopher and psychologist Carl Stumpf to further investigate musical perception (beginning in 1883). To Stumpf, it provided a paradigm for experimental psychology as mental functions and phenomena could be studied in detail. These functions and phenomena are the actual objects of scientific study in Stumpf's inductive and descriptive psychology. Combining insights from statistics, ethnology, anthropology, psychoacoustics and the cultural history of mankind, Stumpf and his team developed a new blend of science which absorbs styles of reasoning, analytical procedures and academic convictions from natural history, the natural sciences and the humanities but at the same time identifies shortcomings of these approaches that fail to grasp the complexities of psychic functions. Despite their reliance on the quasi-objective phonograph and despite their commitment to objectivity, precision and measurement, mental phenomena relating to tonal perception and to music provided too complex a challenge to be easily articulated and shared by the scientific community after 1900. The essay illustrates these tensions against the background of a history of objectivity. PMID- 20027768 TI - Projections of the revolutionary nation: French expeditions in the Pacific, 1791 1803. AB - Revolutionary France's two Pacific expeditions, under the command of Jean-Antoine Bruny d'Entrecasteaux (1792-94) and Nicolas Baudin (1801-1804), demonstrate the importance of scientific inquiry to the newly sovereign nation. France's scientific community adapted to the changed circumstances of revolutionary upheaval by describing its work in terms of national priorities. Individuals on board the expeditions, both naval and scientific personnel, behaved as scientific citizens, intent on composing an encyclopedic body of knowledge about the Pacific. Disputes over whose science mattered more and how credit should be assigned through publication, however, broke down the consensus that science should be a national project. PMID- 20027769 TI - Material experiments: Environment and engineering institutions in the early American republic. AB - In nineteenth-century America, strength of materials, an engineering science, focused on empirical research that yielded practical tools about how to predict the behavior of a wide variety of materials engineers might encounter as they built the nation's infrastructure. This orientation toward "cookbook formulae" that could accommodate many different kinds of timber, stone, mortar, metals, and so on was specifically tailored for the American context, where engineers were peripatetic, materials diverse, and labor in short supply. But these methods also reflected deeper beliefs about the specialness of the landscape and the providential site of the American political experiment. As such, engineers' appreciation of natural bounty both emerged from and contributed to larger values about exceptionalism and the practical character of Americans. PMID- 20027770 TI - The children's republic of science in the antebellum literature of Samuel Griswold Goodrich and Jacob Abbott. AB - The antebellum years in the United States were marked by vigorous debates about national identity in which issues of hierarchy, authority, and democratic values came under intense scrutiny. During this period, a prime objective of indigenous authors writing for American children was educating the young so they would be ready to assume their republican responsibilities. The question of how depictions and discussions about nature and science were deployed toward this end is explored by examining key texts about nature and science from the era's two most prolific and popular children's authors--Samuel Griswold Goodrich (1793-1860) and Jacob Abbott (1803-79)--and highlighting assumptions within these works about what the proper relationship should be between the search for scientific knowledge and the larger polity. PMID- 20027771 TI - "Signs of the times": Medicine and nationhood in British India. AB - Medical practice and research in colonial India historically had been an imperial preserve, dominated by the elite members of the Indian Medical Service. This was contested from the 1900s on by the emerging Indian nationalism. This essay studies debates about the establishment of a medical research institution and how actors imposed the political identities of nationalism on British colonial practices of medical science. At the same time, Indian nationalism was also drawing from other emerging ideas around health and social welfare. The Indian nationalists and doctors sought to build the identities of the new nation and its medicine around their own ideas of its geography, people, and welfare. PMID- 20027772 TI - Informing, involving or engaging? Science communication, in the ages of atom-, bio- and nanotechnology. AB - Science communication has shifted considerably in Europe over the last decades. Three technology controversies on atoms, genes, and nanoscale sciences and nanotechnologies (NST) turned the style of communication from one-way information, participation and dialogues to the idea of an early and more democratic engagement of the public. Analyzing science communication developing over the three controversies, this article shows that what happened in one technology field fed forward to and contributed to shaping the subsequent field and that communication was initiated at a progressively earlier stage of technology development. The article concludes with an empirical analysis of six public engagement projects in NST, saying that the shift towards more democratic engagement of the public hasn't been as profound and complete as has been thought. This is particularly due to the continuing adoption of a simplistic contrast structure that opposes science and the public as two self-contained, antagonistic social entities. PMID- 20027773 TI - "The post-antibiotic apocalypse" and the "war on superbugs": catastrophe discourse in microbiology, its rhetorical form and political function. AB - Discourses evoking an antibiotic apocalypse and a war on superbugs are emerging just at a time when so-called "catastrophe discourses" are undergoing critical and reflexive scrutiny in the context of global warming and climate change. This article combines insights from social science research into climate change discourses with applied metaphor research based on recent advances in cognitive linguistics, especially with relation to "discourse metaphors." It traces the emergence of a new apocalyptic discourse in microbiology and health care, examines its rhetorical and political function and discusses its advantages and disadvantages. It contains a reply by the author of the central discourse metaphor, "the post-antibiotic apocalypse," examined in the article. PMID- 20027774 TI - Bias in the exchange of arguments: the case of scientists' evaluation of lay viewpoints on GM food. AB - Most perspectives on public participation share the notion that dialogues should be open, allowing participants to articulate and evaluate different views and knowledge claims. We hypothesize that participants' evaluation of claims may be biased because participants have a preference for a particular type or source of a claim. This would hamper an open dialogue. We tested the effect of three variables on scientists' evaluation of claims of the general public about GM food: the claim's favorability towards GM food, the phrasing, and the source of the claim. Results are based on a survey-experiment among 73 biotechnology scientists. Biased processing occurred when scientists evaluated claims. Claims that were corresponding with the attitude of the scientists and that were phrased in a cognitive way were evaluated more positively than claims that were contrasting the attitude of the scientists and that were phrased in an affective way. Contrary to our expectation, scientists evaluated claims of the public more positively than claims of experts. PMID- 20027775 TI - Early reports on aquatic animal diseases. PMID- 20027776 TI - Contribution of the Kyivo-Mogylans'ka Academy graduates to the development of science and education in the Eastern Europe. PMID- 20027777 TI - Historical intersections of psychology, religion, and politics in national contexts. AB - Various types of psychology have come into existence in and have been interacting with a plurality of contexts, contexts that have been radically varying in different states or nations. One important factor in the development of psychology has been the multiple relationships to the Christian religion, whether understood as an institution, a worldview, or a form of personal spirituality. The articles in this issue focus on the intertwinements between institutional religion and national political structures and on their influence on developing forms of psychology in four different national contexts: Spain, the Netherlands, the United Kingdom, and the United States. Within these four settings, aspects of the ways in which varying forms of Christian religion coconstituted, facilitated, and shaped psychology, theoretically, practically, and institutionally, are examined. The formative power of the religions was not independent of the relationships between religion and political power, but rather mediated by these. PMID- 20027778 TI - The soul of Spain: Spanish scholastic psychology and the making of modem subjectivity (1875-1931). AB - The aim of this article is to provide an approach to the study of the relations between psychology and Roman Catholic Scholasticism in the making of Spain as a modern nation-state. The crucial period in this process-extending from the beginning of King Alfonso XII's reign in 1875 to the proclamation of the Second Spanish Republic in 1931-is considered. Attention is focused on Ethics textbooks published by Spanish Scholastic authors throughout the period. Through these school manuals, young students were trained in the ideas of citizenship and social coexistence held by the Catholic Church. An analysis of these didactic, programmatic works shows the central role played by the theory of faculties and modern psychological technologies (psychopedagogy, psychopathology, psychotechnics) in the Scholastic outlook. Thus, an attempt is made to show that psychology was used by Spanish Scholasticism as a way of legitimating a reactionary view of Spain, which eventually led to the emergence of National Catholicism as the official ideology of the Franco regime (1939-1975). PMID- 20027779 TI - Ideology, politics, and personality: shaping forces in Dutch psychology of religion, 1907-1957. AB - Although the academic establishment of the psychology of religion in the Netherlands has been stronger than in any other Western country, the start of these developments has been remarkably late (in 1957), especially when taking into account that Dutch academic life: (1) before World War II modeled itself after Germany (where psychology of religion flourished); and (2) was to a considerable extent included in the system of pillarization, which characterized Dutch society at large. The general factors that can be distinguished as having played an important role in the shaping of the situation for psychology of religion in the Netherlands had different impacts in the several universities under consideration. PMID- 20027780 TI - Goodwill in the valley of the shadow: religion, psychotherapy, and politics in Britain from 1945 to the early 1960s. AB - The author hypothesizes that the British psychological climate following World War II, from 1945 to approximately 1960, created the conditions for an unusually frank, honest, and collaborative debate on matters related to religion and spirituality generally as well as broader social and political issues. The author first sketches the nature of this climate, before looking more specifically at the interrelationships among religion, psychotherapy and, very briefly, politics during this period. Suggestions regarding the dissipation of this phase after 1960 are examined. The final section considers some qualifications to the case previously outlined and identifies issues and questions that the author feels need addressing further. PMID- 20027781 TI - Out of the ghetto: integrating Catholics into mainstream psychology in the United States after World War II. AB - The American Catholic Psychological Association (ACPA) was a voluntary association that formed and then transformed itself during a distinctive period of American history. Socially, American Catholics were primed to emerge from what they called their "ghetto," as this formerly largely immigrant group began to enter the economic and social mainstream. Institutions of higher education and psychology were recipients, moreover, of federal funding in the wake of World War II, and some of this money flowed to Catholic institutions. The ACPA began in 1947 as a way to bring Catholics in greater numbers into psychology and also to bring a Catholic perspective to bear on psychology. This article describes and analyzes the major initiatives of the ACPA: the establishment of undergraduate and graduate programs in psychology at Catholic colleges; placement services for members; and the development of psychological assessment programs for candidates for the religious life. In these ways, empirical psychological categories became part of Catholic discourses about psychological life. The ACPA voluntarily restructured itself in the late 1960s in response to a changed social environment and to a consensus that its first aim had been achieved. New issues were surfacing, making a denominational group irrelevant. The ACPA became Psychologists Interested in Religious Issues (PIRI) in 1970. PMID- 20027782 TI - Capitalist contexts for Darwinian theory: land, finance, industry and empire. AB - When socio-economic contexts are sought for Darwin's science, it is customary to turn to the Industrial Revolution. However, important issues about the long run of England's capitalisms can only be recognised by taking a wider view than Industrial Revolution historiographies tend to engage. The role of land and finance capitalisms in the development of the empire is one such issue. If we historians of Darwin's science allow ourselves a distinction between land and finance capitalisms on the one hand and industrial capitalism on the other; and if we ask with which side of this divide were Darwin and his theory of branching descent by natural selection aligned, then reflection on leading features of that theory, including its Malthusian elements, suggests that the answer is often and largely, though not exclusively: on the land side. The case of Wallace, socialist opponent of land capitalism, may not be as anomalous for this suggestion as one might at first think. Social and economic historians have reached no settled consensuses on the long-run of England's capitalisms. We historians of Darwin's science would do well to import some of these unsettled states of discussion into our own work over the years to come. PMID- 20027783 TI - The return of the phoenix: the 1963 International Congress of Zoology and American zoologists in the twentieth century. AB - This paper examines the International Congress of Zoology held in Washington D.C. in 1963 as a portrait of American zoologists' search for effective and rewarding relationships with both each other and the public. Organizers of the congress envisioned the congress as a last ditch effort to unify the disparate subdisciplines of zoology, overcome the barriers of specialization, and ward off the heady claims of more reductionist biologists. The problems zoologists faced as they worked to fulfill these ambitious goals illuminate some of the challenges faced by members of the naturalist tradition as they worked to establish disciplinary unity while seeking public support in the competitive world of twentieth century science. PMID- 20027784 TI - Sources of Wilhelm Johannsen's genotype theory. AB - This paper describes the historical background and early formation of Wilhelm Johannsen's distinction between genotype and phenotype. It is argued that contrary to a widely accepted interpretation (For instance, W. Provine, 1971. The Origins of Theoretical Population Genetics. Chicago: The University of Chicago Press; Mayr, 1973; F. B. Churchill, 1974. Journal of the History of Biology 7: 5 30; E. Mayr, 1982. The Growth of Biological Thought, Cambridge: Harvard University Press; J. Sapp, 2003. Genesis. The Evolution of Biology. New York: Oxford University Press) his concepts referred primarily to properties of individual organisms and not to statistical averages. Johannsen's concept of genotype was derived from the idea of species in the tradition of biological systematics from Linnaeus to de Vries: An individual belonged to a group - species, subspecies, elementary species - by representing a certain underlying type (S. Muller-Wille and V. Orel, 2007. Annals of Science 64: 171-215). Johannsen sharpened this idea theoretically in the light of recent biological discoveries, not least those of cytology. He tested and confirmed it experimentally combining the methods of biometry, as developed by Francis Galton, with the individual selection method and pedigree analysis, as developed for instance by Louis Vilmorin. The term "genotype" was introduced in W. Johannsen's 1909 (Elemente der Exakten Erblichkeitslehre. Jena: Gustav Fischer) treatise, but the idea of a stable underlying biological "type" distinct from observable properties was the core idea of his classical bean selection experiment published 6 years earlier (W. Johannsen, 1903. Ueber Erblichkeit in Populationen und reinen Linien. Eine Beitrag zur Beleuchtung schwebender Selektionsfragen, Jena: Gustav Fischer, pp. 58-59). The individual ontological foundation of population analysis was a self-evident presupposition in Johannsen's studies of heredity in populations from their start in the early 1890s till his death in 1927. The claim that there was a "substantial but cautious modification of Johannsen's phenotype genotype distinction" (Churchill, 1974, p. 24) from a statistical to an individual ontological perspective derives from a misreading of the 1903 and 1909 texts. The immediate purpose of this paper is to correct this reading of the 1903 monograph by showing how its problems and results grow out of Johannsen's earlier work in heredity and plant breeding. Johannsen presented his famous selection experiment as the culmination of a line of criticism of orthodox Darwinism by William Bateson, Hugo de Vries, and others (Johannsen, 1903). They had argued that evolution is based on stepwise rather than continuous change in heredity. Johannsen's paradigmatic experiment showed how stepwise variation in heredity could be operationally distinguished from the observable, continuous morphological variation. To test Galton's law of partial regression, Johannsen deliberately chose pure lines of self-fertilizing plants, a pure line being the descendants in successive generations of one single individual. Such a population could be assumed to be highly homogeneous with respect to hereditary type, and Johannsen found that selection produced no change in this type. Galton, he explained, had experimented with populations composed of a number of stable hereditary types. The partial regression which Galton found was simply an effect of selection between types, increasing the proportion of some types at the expense of others. PMID- 20027786 TI - Germs and Jim Crow: the impact of microbiology on public health policies in progressive era American South. AB - Race proved not merely a disadvantage in securing access to prompt and appropriate medical care, but often became a life and death issue for blacks in the American South during the early decades of the twentieth century. This article investigates the impact some of the new academic disciplines such as anthropology, evolutionary biology, racially based pathology and genetics had in promoting scientific racism. The disproportionately high morbidity and mortality rates among blacks were seen as a consequence of inherent racial deficiencies that rendered any attempt to ameliorate their situation as futile. While the belief in a different pathology in blacks initially deterred most health officials from taking any action, advances in medicine and microbiology, in particular the germ theory, stirred a variety of responses out of sheer self preservation, as fears among whites at the first sign of an epidemic initiated sporadic and limited actions. Ironically, in an era of deepening scientific racism, public health initiatives based on a better understanding of disease causing microorganisms, gradually improved black health. However, some public health measures were hijacked by eugenicists and racists and, rather than addressing the ill health of blacks, public health policy complied with the new laws of heredity by promoting drastic measures such as involuntary sterilization or even abortion. This further complicated the strained relationship between southern blacks and health care professionals and effected ongoing distrust towards public healthcare services. PMID- 20027785 TI - The laboratory technology of discrete molecular separation: the historical development of gel electrophoresis and the material epistemology of biomolecular science, 1945-1970. AB - Preparative and analytical methods developed by separation scientists have played an important role in the history of molecular biology. One such early method is gel electrophoresis, a technique that uses various types of gel as its supporting medium to separate charged molecules based on size and other properties. Historians of science, however, have only recently begun to pay closer attention to this material epistemological dimension of biomolecular science. This paper substantiates the historiographical thread that explores the relationship between modern laboratory practice and the production of scientific knowledge. It traces the historical development of gel electrophoresis from the mid-1940s to the mid 1960s, with careful attention to the interplay between technical developments and disciplinary shifts, especially the rise of molecular biology in this time-frame. Claiming that the early 1950s marked a decisive shift in the evolution of electrophoretic methods from moving boundary to zone electrophoresis, I reconstruct various trajectories in which scientists such as Oliver Smithies sought out the most desirable solid supporting medium for electrophoretic instrumentation. Biomolecular knowledge, I argue, emerged in part from this process of seeking the most appropriate supporting medium that allowed for discrete molecular separation and visualization. The early 1950s, therefore, marked not only an important turning point in the history of separation science, but also a transformative moment in the history of the life sciences as the growth of molecular biology depended in part on the epistemological access to the molecular realm available through these evolving technologies. PMID- 20027787 TI - Edward Hitchcock's pre-Darwinian (1840) "tree of life". AB - The "tree of life" iconography, representing the history of life, dates from at least the latter half of the 18th century, but evolution as the mechanism providing this bifurcating history of life did not appear until the early 19th century. There was also a shift from the straight line, scala naturae view of change in nature to a more bifurcating or tree-like view. Throughout the 19th century authors presented tree-like diagrams, some regarding the Deity as the mechanism of change while others argued for evolution. Straight-line or anagenetic evolution and bifurcating or cladogenetic evolution are known in biology today, but are often misrepresented in popular culture, especially with anagenesis being confounded with scala naturae. Although well known in the mid 19th century, the geologist Edward Hitchcock has been forgotten as an early, if not the first author to publish a paleontologically based "tree of life" beginning in 1840 in the first edition of his popular general geology text Elementary Geology. At least 31 editions were published and those between 1840 and 1859 had this "paleontological chart" showing two trees, one for fossil and living plants and another for animals set within a context of geological time. Although the chart did not vary in later editions, the text explaining the chart did change to reflect newer ideas in paleontology and geology. Whereas Lamarck, Chambers, Bronn, Darwin, and Haeckel saw some form of transmutation as the mechanism that created their "trees of life," Hitchcock, like his contemporaries Agassiz and Miller, who also produced "trees of life," saw a deity as the agent of change. Through each edition of his book Hitchcock denounced the newer transmutationist hypotheses of Lamarck, then Chambers, and finally Darwin in an 1860 edition that no longer presented his tree-like "paleontological chart." PMID- 20027788 TI - [Early achievements of the Danish pharmaceutical industry--2. The minor and almost forgotten pharmaceutical companies]. AB - The article series provides an account in words and pictures of the Danish pharmaceutical industry's products from the earliest times until about 1950. Part 2 deals with products from 16 minor pharmaceutical companies, founded in the last decades of the 19th century and the first decades of the 20th century. Mentioned in chronological order, according to year of foundation, the companies are: C.R. Evers & Co., Jensen & Langebek-Petersens chemisk-techniske Fabrik, Leerbeck & Holms kemiske Fabriker, A/S Skelskor Frugtplantage, Fabriken Ferrin, Chr. F. Petri, Erslevs kemiske Laboratorium, A/S Edward Jacobsen, Th. Fallesen-Schmidt, Fabriken Ferraton, Chemia, Fabriken Kemisan, Central-Laboratoriet, F.F. Gonget & Co., A/S Ejco, and M. Schultz chemiske Fabrik. None of these minor pharmaceutical companies exist today as independent firms. All of them are either closed down or merged into other firms after a number of years. The bigger pharmaceutical companies ensured their continued existence by research and development of new products. The minor companies were not innovative to the same extent, but they played a role at an early stage in the production of Danish copy medicine and in that way a role in the establishment of a Danish generic pharmaceutical industry. The earliest products included dietetic preparations as malt products and albumin maltose products, and iron preparations, often with an admixture of medicine substances. Real medicines such as sleeping, analgesic and antipyretic medicines as well as anesthetics followed later. PMID- 20027789 TI - Global commercialization of a technology. The multinational journey of Repro-Dose drug delivery system emerging of Benzon Pharma A/S, Copenhagen, Denmark. PMID- 20027790 TI - [Lauritz Toft and DUMEX--the man, the idea and the way to its realization]. AB - Lauritz Toft (1920-1991), also known as Lau, graduated with a MSc degree in pharmacy at the Royal Danish School of Pharmacy in 1944. During the education and parallel activities Lau had shown gifts for intuition and improvisation, together with special talents for leadership and large-scale working. In 1945 he got the idea during the organization of the East Asiatic Company Ltd. to sell the best of the products from the Danish pharmaceutical companies in India under a common trade mark: DUMEX (Danish United Medical Export). The article describes Lau's difficulties and problems in realizing this idea. The adventure peaks in the mid 1950's with about 40 pharmacists in DUMEX-departments in India as well as Thailand, Malaysia, Indonesia, China etc. PMID- 20027791 TI - [On the history of pharmacy in Indochina (1861-1954)]. AB - The French episode in Indochina starts in the beginning of the 1860's with Saigon's capture (1861) and the start of the protectorat for Cambodia. As in the other countries that became part of the french colonial empire, military and civilian pharmacists will be very early on part of the organization of sanitary activities and the valorisation of local resources. Although local pharmacies were maintained through out the entire colonial period, occidental pharmacies were installed very soon, Lourdeau's pharmacy being implemented as early as 1865 in Saigon where he was the mayor. Military pharmacists will also be present, their activities being often far from pharmacist activities, as for example Victor-Alphonse Massie in Laos, or Joseph Forestier who became indegen affairs administrator in Saigon from 1874 (at the age of 24) up to 1896. Military pharmacists were also a major resource as teachers of pharmacy in Indochine. The abundance of valuable medicinal herbs in the region was an opportunity for pharmacists to enrich pharmacopea. Roux, a military pharmacist, was also the one that sucessfully introduced hevea in Indochine, allowing the development of what remains a valuable industry in Vietnam. PMID- 20027792 TI - [A long-playing record: contraception]. AB - The humoristic record "Contraception", recorded during the year 1976 by the humorist Rene Cousinier, conducted the author of this article to study contraceptives means used in France from 1968 to 1976. Here are studied firts pregnancy tests; the birth of Planning familial and Neuwirth law; each classical contraceptives. The Pill is fully related in detail. Each part is illustrated with citations from the record. PMID- 20027793 TI - [Trehala, a meeting point between zoology, botany, chemistry, and biochemistry]. AB - Trehala is a crude drug consisting of the pupal chambers formed by insects belonging to the genus Larinus that live on several Echinops species of the Middle-East. This sweet cocoon is locally used as human food and also for the treatment of cough and various pulmonary diseases. It first appeared in Western Europe in the collection of drugs from the Ottoman Empire displayed by Francois Della Sudda during the International Exhibition held in Paris in 1855. On the basis of this sample Nicolas Guibourt (1790-1867) gave, in 1858, the first full scientific description of the drug, its origin, and Larinus nidificans as the main insect species responsible for its formation. Marcellin Berthelot (1827 1907) isolated in the same year the sugar trehalose from the drug and gave a full account of its physical and chemical properties. In 1876, Muntz established that trehalose was identical with mycose isolated from Claviceps purpurea by Mitscherlich. PMID- 20027794 TI - [Historical aspects of some infectious diseases: is their gradual eradication utopian?]. AB - Infectious microorganisms (parasites, bacteria, viruses) which caused dramatic epidemics in human populations throughout the centuries, as well as their natural reservoirs and possible vectors, have not disappeared... Plague, cholera, malaria, tuberculosis, rickettsioses are still quite present in some areas of the world. Furthermore new infectious diseases caused by emerging pathogens have recently appeared dramatically, such as AIDS. Eradication of the most severe and lethal infectious diseases remains nevertheless an ambitious but not utopian objective, if one considers the spectacular advances made, more or less recently, in the field of specific vaccinations. For instance, smallpox has been eradicated around 1980. Present efforts are mainly concentrated on poliomyelitis, the eradication of which was hoped to take place in year 2005, but which is still present in a few areas. Eradication of measles, with the presently available vaccine, is a reasonable objective. Eradication of malaria appears to be much more difficult since this infection involves a complex parasite and an insect vector, the mosquito. And, as far as AIDS is concerned, intensive research has not yet led to the conception of an effective vaccine. PMID- 20027795 TI - [Leon Monsel and his solution]. AB - Leon Monsel was born on March 13, 1816 in Gironde. He did training courses in chemist's first near of home and then in Paris. He also studied pharmacy in Pharmacy School of Paris where he succeeded on August 12, 1840 and he entered army. He remained in it until his retirement in 1874. He died at Nancy on April 15, 1878. Monsel is overall well known by his publications about coagulation power of ferric sub-sulfate from which he described three preparation ways. Current publications are exclusively foreigner. They point out advantages and disadvantages of Monsel's solution. It is used to day as hemostatic agent in minor surgical procedures such as biopsies in genecology, dermatology, proctology and O.R.L. Besides when he was stationed in Roma he studied Acqua Acetosa (acid salty gaseous water) of this town and later when he was stationed in Bordeaux, he took out a patent about manufacturing varnish from metallic resinates. PMID- 20027796 TI - Herbal medicine, what physicians need to know. AB - Herbal medicine, the most major component of traditional medicine, is as old as recorded history. Beginning in the early 1800s, with the development in the science of chemistry, a new era in pharmacotherapeutics was initiated whereby active chemical ingredients in plants, historically known to produce a favorable therapeutic effect, were extracted, purified and their structure disclosed. This ushered the modern era of therapy with drugs based on exploration of pure chemical products as to chemical identity, physicochemical properties, pharmacodynamic actions, pharmacokinetic behavior in the biological system, toxicological profile and effective and safe application in therapy. This relegated herbal medicine to a secondary role. More recently, a revival in the use of herbal medicine has been witnessed, even in culturally advanced societies, probably enhanced by the false belief that natural products are safe and also by vigorous promotion. Parallel to the increase in the use of herbal preparations as remedies for major diseases, there is currently a growing concern about their efficacy, safety and control. This prompted the World Health Organization to come out with recommendations for control in the document "Research Guidelines for Evaluating the Safety and Efficacy of Herbal Medicines" in 1993. The guidelines are equal in strictness to those applicable for drugs in general. A large number of member states have adopted these guidelines. The dangers in using herbal preparations for treatment include: * unproven therapeutic benefit * undisclosed toxicities * interaction of the chemicals in herbal preparations with each other and with concomitantly taken drugs, at the level of functionally important biological entities such as the plasma proteins, receptors, ion channels, transporters and others * incompatibilities with patient-related factors such as age, sex, genetic background and the function of the organs responsible for eliminating the effects of chemicals in herbal preparations such as the liver or kidney * the difficulty in standardizing treatments and * the inclusion of regular drugs with the herbal preparations without disclosure. In Lebanon, a committee is charged with the control of herbal preparations but a stricter control is required to handle preparations with a therapeutic claim as well as the issue of promotion. PMID- 20027797 TI - [Autoimmune thyroid disease. Clinical and biological correlations]. AB - In this article, we analyze the clinical and biological data concerning the autoimmune thyroid diseases in patients recruited in an endocrinology clinic at the university hospital center of Hotel-Dieu de France between March 2005 and November 2005. We studied 121 patients (51 with Basedow disease and 70 with Hashimoto thyroiditis), between 13 and 68 years old, with a BMI of 24.68 kg/m2 and with a female predominance (105 women). Symptoms of hyperthyroidism represented the most frequent cause of consultation. The distribution of patients regarding their thyroid disease showed that 42.1% of patients had hyperthyroidism (only one patient had subclinical hyperthyroidism), 21.5% had a subclinical hypothyroidism, 28.1% had clinical hypothyroidism and 8.3% had euthyroid goiter. Half of the patients had at least a member of their family whith a thyroid disease. The autoimmune thyroid diseases are strongly associated to other autoimmune diseases and to repetitive spontaneous abortion. Thus, 39% of the married women had had at least one spontaneous abortion and 26.4% of the patients had one or more autoimmune disease associated to their thyroid disease; diabetes mellitus type 1 representing the most frequent one. Concerning the treatment, we remarked a remission of 30 patients (66.7%) with Basedow disease after 18 months of antithyroid drug treatment of 45 patients. In Hashimoto thyroiditis, we remarked a frequent evolution of patients with subclinical hypothyroidism to overt hypothyroidism when medical treatment was not initiated. PMID- 20027798 TI - Impact of a communication skills audiovisual package on medical students' knowledge. AB - INTRODUCTION: Over the last decade more emphasis is being put on teaching communication skills (CS). Use of videos and role-play was suggested to improve CS. This article will present the impact of an audiovisual package on promoting the knowledge of medical students in CS. METHODS: Seventy-five second year medical students--distributed into eight groups led by four facilitators- critiqued a video clip immediately before and after the introduction of a communication skills audiovisual package. The skills taught included opening the interview, questioning, facilitation, clarification, reflection, confrontation, summarizing, and preparation of the patient for the physical exam. The students, also, role-played the reviewed scenario. The students' pre- and post-intervention responses were analyzed using a standardized grading form. RESULTS: There was a significant improvement in students' knowledge (p < 0.000) after the introduction of the intervention in all the CS taught except closed ended questioning. This improvement was consistent among the four facilitators. CONCLUSION: Reviewing video scenarios and role-playing improved the knowledge in core communication skills among second-year medical students assessed by a video-based written examination. PMID- 20027799 TI - [Anatomy of the larynx with 64-channel multidetector CT]. AB - Knowledge of axial anatomy of the different laryngeal structures, identification of the plan of the cords, and relations between the true and false vocal cords is essential for laryngeal surgery where anatomical and radiological correlation is necessary for optimal orientation and guidance. This review constitutes an atlas of axial anatomy of the larynx obtained from a multidetector CT scan, describing the skeletal cartilage, the different folds constituted by laryngeal ligaments covered by mucosa, and the fatty laryngeal spaces. It shows that CT scan and the different reconstructions specifically virtual endoscopy are a valuable adjunct in laryngeal exploration. PMID- 20027800 TI - [Risk factors of permanent functional limitations on 210 hemodialysed patients]. AB - The aim of the study is to identify risk factors that can worsen the disability in chronic hemodialysis patients. PATIENTS AND METHODS: A transversal study was conducted on a total of 210 patients. Data was collected using a 42-item questionnaire, and univariate and multivariate statistical analyses were conducted. RESULTS: Univariate analysis reveals that male sex, advanced age, small stature, low weight, unemployment, lifestyle, lack of sport, hypertension or diabetes, co-morbid conditions, polysulfane filter, time on dialysis, diffuse and chronic pain with high analog visual score are all aggravating disability. Multivariate study confirms advanced age, male sex, hemiplegia, cardiac failure and high analog visual scale as factors aggravating handicap in hemodialysis patients. CONCLUSION: Modifiable and non modifiable factors can be identified in the disability of dialysis patients. The approach for such patients should be targeted and multifactorial with emphasis on the role of physical rehabilitation and exercises. PMID- 20027801 TI - Tobacco use and cessation counseling among health professional students: Lebanon Global Health Professions Student Survey. AB - INTRODUCTION: The number of deaths attributable to tobacco use is rising globally. Health professionals can help reduce tobacco use by providing advice to patients regarding smoking cessation. Very few studies have collected information on tobacco use and cessation counseling training among health professional students. The purpose of this paper is to examine these issues using data from the 2005 Lebanon Global Health Professions Student Survey (GHPSS). METHODS: The 2005 Lebanon GHPSS includes nationally representative estimates of third-year students in all dental, medical, nursing, and pharmacy schools in Lebanon. RESULTS: Current cigarette use ranged from 14.8% for pharmacy students to 26.9% for nursing students and 27.4% for medical students. Current waterpipe use ranged from 20.6% for medical students to 44.9% for nursing students. About 8 in 10 students in all four disciplines were exposed to second-hand smoke in public places. More than 8 in 10 students in medical, nursing, and pharmacy schools believed that health professionals have a role in giving advice or information on smoking cessation to patients. More than 9 in 10 students in all four disciplines believed that health professionals should receive training on smoking cessation techniques. However, those who received training ranged from 19.8% of pharmacy students to 43.7% of nursing students. CONCLUSION: Tobacco use (cigarettes and water-pipe) is high among third-year health professional students in Lebanon. Students are willing to provide smoking-cessation counseling but lack training. The Ministry of Public Health, the Ministry of Education, and educational institutions in Lebanon should work together with other interested partners in developing, testing, and implementing successful patient cessation counseling training programs for health professionals. PMID- 20027802 TI - Focus groups findings reveal barriers to teaching communication skills to medical students. AB - BACKGROUND: In December 2002, 74 medical students in the second preclinical years at the American University of Beirut (AUB) attended a workshop on basic communication skills (CS). The students watched video clips which demonstrated different communication skills and stages of the consultation, and role-played to try out ways they could improve the consultation. Their familiarity with the skills taught was assessed at the beginning of the workshop, immediately afterwards, and again six months later. There was a significant improvement noted immediately after the intervention but this apparent gain was lost at six months. METHOD: Sixteen students, who attended the workshop, also attended two focus group discussions which were aimed to assess student recall of the communications skills workshop, and explore ways in which the students thought the knowledge gained could be maintained. RESULTS: In general students thought that lack of exposure to patients at the time when the CS course was held, and a lack of consistency in CS coupled with poor faculty CS role modeling hindered their ability to maintain and practice good CS. CONCLUSIONS: Teaching CS may be more effective if medical students are taught these skills when they are exposed to patients on a regular basis, and the training is maintained throughout the clinical years. However, more emphasis should be on reminding faculty members of their role as teachers and good communicating role models to students through workshops and seminars. PMID- 20027803 TI - [Daily calcium intake and body mass index in a group of Lebanese adolescents]. AB - AIM OF THE STUDY: To investigate the relationship between daily calcium intake (DCI) and body mass index (BMI) in a group of Lebanese adolescents. METHODS AND RESULTS: 419 adolescents (219 boys and 169 girls) aged 13-18 years participated in our study. DCI was calculated using a validated questionnaire, and anthropometrical characteristics (weight and height) were measured. In this study, only 20% of the adolescents met the adequate DCI recommendation of 1300 mg/day. Boys had a significantly higher mean DCI than girls (1023 +/- 360 mg/d and 839 +/- 303 mg/d respectively for boys and girls ; p < 0.001). DCI was negatively related to BMI in boys (r = -0,15; p < 0.05) but not in girls. Obese boys had a significantly lower mean DCI than those whose BMI was normal (869 +/- 249 mg/d and 1043 +/- 373 mg/d respectively ; p < 0.05). CONCLUSION: In this study, 80% of the adolescents do not meet the DCI adequate intake. It seems important to encourage these adolescents to increase their DCI. Moreover, this study shows that DCI is inversely related to age in girls and to BMI in boys. PMID- 20027804 TI - [Skin and eating, their relationship to myths]. AB - When eating certain foods, one also eats some of its imaginary, physical or moral characteristics by analogy. The skin does not escape this "eating magic", since the skin's appearance is said to mirror one's self and above all one's nutrition. Since pleasure implies transgression, dieting exists to punish and reestablish taboos but also to insure redemption. Patients often distrust certain food that allegedly "damage one's complexion" or "provoke redness, spots and itching". Lists of such foods vary amongst patients, but share common features: these foods are "hot", in Arabic Hami. Such concepts, common currency within the Lebanese population, are residues of Aristotle and Hippocrates's medical categorizations. They were developed through centuries by Arab physicians, and remain nowadays among Middle-Easterners as dietary restrictions only. Some physicians therefore feel free to prohibit foods that adolescents are particularly fond of, especially when treating acne. Nutrition has an undeniable importance in a single domain- food allergies. These allergies, frequent amongst infants, are much rarer in adult life. It is thus important to carefully examine patients and investigate their cases in order to determine whether alleged allergies are real and to diagnose them correctly. PMID- 20027805 TI - Xanthophylls and eye health of infants and adults. AB - Lutein and zeaxanthin are the only carotenoids present in the eye. They cannot be synthesized de novo and are specifically concentrated in the macula. They appear to have at least two major functions: to filter out blue light and thus prevent ensuing damages to the eye and to act as antioxidants. Infants are particularly at risk from both blue light and oxidative damage to eye tissues. Lutein is present in human milk but is not currently added to infant formulas. Fortifying formulae with lutein in order to match more closely human milk might help protect the infant's sensitive eyes. In adults, the exact pathogenesis of age-related maculopathy remains unknown. Light damage, inflammation, and the disruption of cellular processes by oxidative stress may play an important role in the degenerative process. Manipulation of intake of xanthophylls has been shown to augment macular pigment, therefore it is thought that carotenoid dietary supplements could prevent, delay, or modify the course of age-related maculopathy. However, definite evidence of the effect of carotenoids, the optimal doses to use, and the supplementation duration are still under investigation. PMID- 20027806 TI - Epidermoid cyst of the spleen: case report. AB - Splenic cysts, very rare pathologies, are classified into parasitic and the highly uncommon, non parasitic cysts. Based on the presence or absence of an epithelial lining wall, the latters are classified into true cysts and false cysts. We present a case of a 23-year-old male who presented to our clinic with a several-month history of abdominal discomfort that was due to the compressive effect of a huge nonparasitic true splenic cyst. PMID- 20027807 TI - Migration in two directions. AB - BACKGROUND: Percutaneous endoscopic gastrostomy (PEG), the modality of choice for long-term enteral access, is generally a safe procedure but can be associated with many potential complications. OBJECTIVES: Report two different and late complications of PEG in two patients fed at home, leading them to the emergency department. CASE REPORT: A 75-year-old man and a 14-year-old young man with PEG presented to the emergency department with two different complications related to the gastrostomy tube. The first patient developed fever and deterioration in mental status due to parietal abscess which developed secondary to the migration of the internal button of the gastrostomy tube in the abdominal wall. He was treated with antibiotics and the gastrostomy tube was extracted. The second one presented upper gastrointestinal bleeding due to intestinal perforation at the level of the internal button of the gastrostomy tube. Bleeding and perforation were treated conservatively and he had a good evolution. CONCLUSION: Persons taking care of patients with PEG tube must be aware of potential complications. The position and the permeability of the tube must be systematically checked before feeding and medical advice should immediately be asked for in case of doubt or in the presence of any alarming sign. PMID- 20027809 TI - Evaluation of the usefulness of a simple touch-panel method for the screening of dementia. AB - In clinical settings, Hasegawa's dementia scale, revised (HDS-R), and the mini mental state examination (MMSE) are widely employed as simple mental function tests useful for the diagnosis of dementia. In recent years, for the early diagnosis of dementia, a simple computerized touch panel-type screening test (touch panel-type screening test), called the "forgetfulness consultation program" (Nihon Kohden Corp.), has been developed. We performed dementia screening using this touch panel-type screening test in 95 elderly subjects, and evaluated its usefulness in comparison with HDS-R or MMSE. The results of evaluation using the touch panel-type screening test were significantly correlated with those using HDS-R and MMSE in the elderly subjects. This touch panel-type screening test was not time-consuming (about 3 min) since it includes only a small number of test items. It could also be performed solely by the examinee, and so was free from examiner-related bias. Therefore, this method may be very useful for the diagnosis of dementia and evaluation of its severity. PMID- 20027808 TI - NSAID-induced colopathy: case report and review of the literature. AB - Non-steroidal anti-inflammatory drugs (NSAIDs) are widely prescribed in different conditions. NSAIDs are well known to induce gastroduodenal damage as well as more distal small bowel and large bowel damage. NSAID-colopathy (ulceration or stricture formation) usually involves the right colon, but the rectum may be involved too. Patients may present with diarrhea, gastrointestinal bleeding, anemia, weight loss or obstruction. Inflammatory bowel diseases and malignancy must be ruled out before establishing the diagnosis. Discontinuation of the NSAID is the rule. Balloon dilatation has been shown to be effective in the treatment of colonic and ileo-colonic strictures. Surgery is reserved for multiple strictures or complications. PMID- 20027810 TI - Ruptured vertebral artery dissecting aneurysms treated with endovascular treatment. AB - Intracranial vertebral artery (VA) dissecting aneurysms often present with severe subarachnoid hemorrhage (SAH) and a subsequent high rate of mortality. We retrospectively studied the treatment efficacy and outcomes of 26 patients with ruptured VA dissecting aneurysms treated with endovascular surgery. Twenty-six patients with ruptured VA dissecting aneurysms were enrolled. Fifteen patients presented with poor-grade SAH and eleven with good-grade SAH. All patients were treated with endovascular treatment. We reviewed modes of therapy, complications and clinical outcomes. Coil occlusion of the artery at the dissecting aneurysm was performed in 24 patients. Proximal parent artery occlusion was performed in 2 patients. Obliteration of the dissecting aneurysm on postoperative angiogram was found in all patients. No patient sustained permanent complications associated with endovascular treatment. Regarding clinical outcome, although five patients died due to severe SAH, 19 patients had a good recovery or moderate disability. Ruptured VA dissecting aneurysms can be managed safely with coil occlusion of the lesion and/or parent artery. PMID- 20027811 TI - The prevalence of urinary lithiasis in children in Van Region, Turkey. AB - We aimed to determine the prevalence and etiology of urinary lithiasis in childhood in our region, Van, Turkey. A total of 1120 children were included in the study. Urinary ultrasonography was performed in all the children between April 2003 and June 2003. During the study, the kidneys were examined longitudinally and transversally using a 3.75 MHz convex probe in a Hitachi EUB 315 ultrasonography machine. The children's ages ranged from 7 years to 14 years (10.16 +/- 1.92 years) and 572 (51.1%) were males and 548 (48.9%) females. Urinary ultrasonography showed that 19 (1.7%) children had urinary lithiasis, which was in the right kidney in 15 children and in the left kidney in four children. Urinary lithiasis was in the upper urinary tract in all children. The etiological studies showed metabolic disorder in 14 children, and congenital renal anomaly in one child, but no underlying cause was diagnosed in four children. In conclusion, we found that was the prevalence of urinary lithiasis was 1.7% in school-aged children in our region. It was also noted that all urinary lithiasis was in the upper urinary system and its most common cause was metabolic disorder. PMID- 20027812 TI - America's best hospitals. PMID- 20027813 TI - Stop a killer before it's too late. PMID- 20027814 TI - Information over exaggeration. PMID- 20027815 TI - Get ready to take charge. PMID- 20027816 TI - Quenching the slow burn. The evidence is mounting that chronic inflammation plays a role in diseases from cancer to heart disease to Alzheimer's. Here's how you can stop it. PMID- 20027817 TI - The diet of the future: geared to your genes? PMID- 20027818 TI - In a workout rut? Exercise a fantasy. PMID- 20027820 TI - How to get the very best cancer care. PMID- 20027819 TI - Getting a good diagnosis. PMID- 20027821 TI - Six smart ways to treat heart failure. PMID- 20027822 TI - Could a dose of stem cells heal a failing heart? PMID- 20027823 TI - Managing your pain, minus addiction. PMID- 20027824 TI - Patients making their voices heard. PMID- 20027825 TI - Should you join a research study? PMID- 20027826 TI - If the gloom won't lift. PMID- 20027827 TI - Battling insomnia? Consider therapy. PMID- 20027828 TI - Broken bond: when a child hates a parent. PMID- 20027829 TI - The lure of an ADHD treatment minus the meds. PMID- 20027831 TI - The power of tracing your medical roots. PMID- 20027830 TI - Creating an online record. PMID- 20027832 TI - Your future health plan. PMID- 20027833 TI - A rash? Where? Show me on the screen. PMID- 20027834 TI - Is a 'cash only' or 'direct pay' doctor right for you? PMID- 20027835 TI - America's best health insurance plans. PMID- 20027836 TI - The Honor Roll. PMID- 20027837 TI - The 'reform' that ate America. PMID- 20027838 TI - A heavy footprint. Strong stance on environment, healthcare urged. PMID- 20027839 TI - Labor's lead. Unions find H1N1 issues give them advantages. PMID- 20027840 TI - Preventive care. Healthcare reform should tackle conflicts of interest, too. PMID- 20027841 TI - Lessons from the Hoosier State. Medicaid expansion might offer glimpse at reform. PMID- 20027842 TI - Heartening trends. Thomson Reuters' 100 top hospitals: cardiovascular benchmarks, 2009. PMID- 20027843 TI - Virtual realities. While 3-D diagnostic imaging has gained ground among providers, cost and reimbursement issues have hindered wider adoption. PMID- 20027844 TI - Another U.S.-Israeli friendship. Patient-safety collaboration seeks broader Middle East participation. PMID- 20027845 TI - Aviation-related injury morbidity and mortality: data from U.S. health information systems. AB - INTRODUCTION: Information about injuries sustained by survivors of airplane crashes is scant, although some information is available on fatal aviation related injuries. Objectives of this study were to explore the patterns of aviation-related injuries admitted to U.S. hospitals and relate them to aviation deaths in the same period. METHODS: The Healthcare Cost and Utilization Project (HCUP) Nationwide Inpatient Sample (NIS) contains information for approximately 20% of all hospital admissions in the United States each year. We identified patients in the HCUP NIS who were hospitalized during 2000-2005 for aviation related injuries based on the International Classification of Diseases, 9th Revision, codes E840-E844. Injury patterns were also examined in relation to information from multiple-cause-of-death public-use data files 2000-2005. RESULTS: Nationally, an estimated 6080 patients in 6 yr, or 1013 admissions annually (95% confidence interval 894-1133), were hospitalized for aviation related injuries, based on 1246 patients in the sample. The average hospital stay was 6.3 d and 2% died in hospital. Occupants of non-commercial aircraft accounted for 32% of patients, parachutists for 29%; occupants of commercial aircraft and of unpowered aircraft each constituted 11%. Lower-limb fracture was the most common injury in each category, constituting 27% of the total, followed by head injury (11%), open wound (10%), upper extremity fracture, and internal injury (9%). Among fatalities, head injury (38%) was most prominent. An average of 753 deaths occurred annually; for each death there were 1.3 hospitalizations. CONCLUSIONS: Aviation-related injuries result in approximately 1000 hospitalizations each year in the United States, with an in-hospital mortality rate of 2%. The most common injury sustained by aviation crash survivors is lower limb fracture. PMID- 20027846 TI - Aortic function in rats after decompression without ultrasonically detectable bubble formation. AB - BACKGROUND: Several studies have demonstrated an adverse effect of bubbles on endothelial function. The degree of dysfunction appears to be related to the number of bubbles present. The aim of the study was to determine whether decompression without bubble formation visible by ultrasound had any effect on arterial endothelial function. METHODS: We decompressed 21 Sprague-Dawley rats weighing 215-260 g from 700 kPa (approximately 6.9 ATA) in a dry hyperbaric chamber followed by a 1-h observation period and measured aortic endothelial dependent relaxation to acetylcholine. Later, we determined the specific weight of the brain as a measure of edema formation and vascular bubbles in the arterial circulation. RESULTS: No bubbles were seen in the pulmonary arteries of seven rats. We found a significant lower vasodilatory response to acetylcholine in the decompressed rats (44% +/- 14%) compared to the control rats (58% +/- 12%) as a sign of endothelial dysfunction. There was no significant difference between the two groups in the specific gravity of the brain. CONCLUSION: We conclude that measurable arterial dysfunction in the aorta can occur even if no visible venous bubble formation is seen. There are no results in this study suggesting that these rats had damaged blood-brain barriers or brain edema. PMID- 20027847 TI - Hyperventilation in response to progressive reduction in central blood volume to near syncope. AB - INTRODUCTION: There is little evidence to support the usefulness in monitoring respiration during casualty triage and transport as an early indicator of hemorrhage severity and trauma patient outcome. We, therefore, tested the hypothesis that hyperventilation can be elicited by progressive reductions in central blood volume independent of metabolic stimuli. METHODS: Progressive central hypovolemia was induced in 10 healthy subjects (5 men, 5 women) by applying lower body negative pressure (LBNP). The LBNP protocol consisted of a 5 min controlled rest period (0% LBNP) followed by progressive 5-min chamber decompressions until the onset of hemodynamic decompensation (LBNP(max)). During each LBNP stage, total minute ventilation volume (V(E)), tidal volume (V(T)), respiratory rate, oxygen uptake (Vo2), end-tidal CO2 (E(T)co2), arterial oxygen saturation (S(p)o2), and venous blood pH and lactate were measured. RESULTS: Compared with baseline, Vo2, S(p)o2, Po2, Pco2, pH, and lactate were unaltered throughout LBNP. V(E) was unaltered through 80% of LBNP tolerance, but increased by 54% during LBNP(max) as a result primarily of elevated V(T), while E(T)Co2 was reduced. CONCLUSIONS: Increased V(E) at LBNP(max) combined with reduced E(T)co2 in the absence of changes in blood and systemic metabolic stimuli support the hypothesis that severe reductions in central blood volume drive hyperventilation. The endogenous "respiratory pump" may be a protective strategy to optimize cardiac filling in conditions of central hypovolemic hypotension, but its late appearance indicates that respiratory parameters may not be useful as a clinical metric for early prediction of patient outcome during hemorrhage. PMID- 20027848 TI - Bed rest and cognition: effects on executive functioning and reaction time. AB - INTRODUCTION: Executive functions are high-order aspects of cognition heavily dependent upon the prefrontal cortex. Both prefrontal cortex activity and executive function task performance are enhanced by participation in aerobic physical activity, suggesting that a lack of such activity during the bed rest model of prolonged weightlessness might induce executive function deficits. METHODS: Twenty-four healthy males (ages 21-45 yr) undertook 60 d of head-down bed rest (-6 degrees) for the 2nd Berlin Bed Rest Study (BBR2-2). Three executive function tasks (Iowa Gambling Task, working memory, and flanker) and a reaction time task were administered before, during, and after bed rest. RESULTS: Iowa Gambling Task scores were significantly worse during bed rest (1.7 +/- 6.9) than in other sessions (24.3 +/- 7.8). Effects on working memory and flanker task performance were less obvious, requiring practice effects to be considered. Reaction time was significantly slower after bed rest (569 +/- 42 ms) than in earlier tests (529 +/- 45 ms). There was also significantly less intrasubject variability in reaction time after bed rest, consistent with more efficient executive functioning at this stage. DISCUSSION: Our results provide some evidence for a detrimental effect of bed rest on executive functioning. Whether this stems from a lack of aerobic physical activity and/or changes in the prefrontal cortex remains to be determined. Cognitive effects of bed rest could have implications for the planned human exploration of Mars, and for medical and lifestyle conditions with inadequate levels of aerobic physical activity. PMID- 20027849 TI - Whole-body vibration effects on bone mineral density in women with or without resistance training. AB - INTRODUCTION: Whole-body vibration exposure may translate into improved bone mass in young adult women. The primary focus of this study was to examine the effects of graded whole-body vibration or vibration exposure plus resistance training on bone mineral density (BMD), hematological measures for bone remodeling, and exercise metabolism in young women. METHODS: There were 51 healthy active women [mean (SD) age, 21.02 (3.39) yr; height, 165.66 (6.73) cm; body mass 66.54 (13.39) kg] who participated in the intervention. Subjects were randomly assigned to whole-body vibration (WBV), whole-body vibration plus resistance training (WBV+RT), or control (CONT) groups for 16 wk. RESULTS: A repeated-measure ANOVA found no significant (P < 0.05) group differences in BMD at the completion of 16 wk. A significant within group change was apparent for the WBV (2.7% femoral neck) and WBV+RT (femoral neck 1.9%; vertebra 0.98%). WBV and WBV+RT experienced a significant (P < 0.05) 60% and 58% increase in adiponectin, 48% and 30% in transforming growth factor-beta1, and 17% and 34% in nitric oxide with an accompanying 50% and 36% decrease in osteopontin, 19% and 34% in interleukin 1beta, and 38% and 39% in tumor necrosis factor-alpha. CONCLUSIONS: The results indicate graded whole-body vibration exposure may be effective in improving BMD by increasing bone deposition while also decreasing bone resorption. Whole-body vibration may also provide an efficient stratagem for young women to achieve peak bone mass and help stave off osteoporosis later in life and provide a novel form of physical training. PMID- 20027850 TI - Physiologic +Gz tolerance responses over successive +Gz exposures in simulated air combat maneuvers. AB - INTRODUCTION: Fighter aircraft pilots are exposed to repetitive headward acceleration (+Gz) during air combat maneuvering. The objective of this study was to compare physiologic responses and the calculated +Gz tolerances of multiple successive +Gz exposures with the responses of the first +Gz exposure. METHODS: There were 13 subjects who performed simulated air combat maneuvering (SACM) profiles composed of 10 rapid-onset rate +Gz cycles with different combinations of short- or long-duration +Gz plateaus (8 or 20 s) and +Gz pauses (1 or 15 s). +Gz, plateaus were individually set at levels inducing strong physiologic responses while the +Gz pause was set at 1.4 Gz. Head-level systolic pressure, ear opacity, and vision quality were measured. RESULTS: The nadirs of head-level systolic pressure, ear opacity, and visual quality during each +Gz plateau were higher in subsequent cycles compared to the first +Gz cycle. There was an average increase in calculated +Gz tolerance of 0.35 +/- 0.21 Gz following the first +Gz cycle. SACMs with short +Gz pauses produced greater increases in +Gz tolerance than SACMs with long +Gz pauses. DISCUSSION: Cardiovascular and visual responses were improved over the course of successive +Gz cycles, indicating that the risk of +Gz-induced loss of consciousness was not increased beyond the first +Gz cycle. The increase in +Gz tolerance beginning with the second cycle is attributed to a carryover of compensatory responses, primarily vasoconstriction, with possible contribution from greater venous return and baroreflex enhancement. PMID- 20027851 TI - Body sway at sea for two visual tasks and three stance widths. AB - BACKGROUND: On land, body sway is influenced by stance width (the distance between the feet) and by visual tasks engaged in during stance. While wider stance can be used to stabilize the body against ship motion and crewmembers are obliged to carry out many visual tasks while standing, the influence of these factors on the kinematics of body sway has not been studied at sea. METHODS: Crewmembers of the RN Atlantis stood on a force plate from which we obtained data on the positional variability of the center of pressure (COP). The sea state was 2 on the Beaufort scale. We varied stance width (5 cm, 17 cm, and 30 cm) and the nature of the visual tasks. In the Inspection task, participants viewed a plain piece of white paper, while in the Search task they counted the number of target letters that appeared in a block of text. RESULTS: Search task performance was similar to reports from terrestrial studies. Variability of the COP position was reduced during the Search task relative to the Inspection task. Variability was also reduced during wide stance relative to narrow stance. The influence of stance width was greater than has been observed in terrestrial studies. CONCLUSIONS: These results suggest that two factors that influence postural sway on land (variations in stance width and in the nature of visual tasks) also influence sway at sea. We conclude that--in mild sea states--the influence of these factors is not suppressed by ship motion. PMID- 20027852 TI - Pre-dive vibration effect on bubble formation after a 30-m dive requiring a decompression stop. AB - INTRODUCTION: The preconditioning of divers to reduce post-dive decompression sickness (DCS) has gained increased interest in diving medical research over the last few years. The beneficial effects of physical exercise, oxygen breathing, hyperbaric exposure, heat exposure, hyperhydration, or nitroglycerin administration before the dive are only a few examples of ongoing research. In this work, we investigated the effects of pre-dive whole-body vibration on post dive bubble formation. METHODS: Following French Navy standard dive procedures, 14 healthy male military divers performed 2 identical dives 1 wk apart to 30 m of seawater (msw) for 30 min. One of the dives was randomly preceded by a 30-min whole-body vibration session (frequencies 35-40 Hz) 1 h before the dive. Post dive bubbles were measured precordially 30, 60, and 90 min after the dive and were graded according to the Kissman Integrated Severity Score (KISS) protocol, with and without knee flexing. Arterial endothelial function was measured before and after vibration using flow mediated dilation (FMD) measurement. RESULTS: A significant reduction in bubble scores was observed after the "vibration" dive. CONCLUSION: As there was no observed change in FMD after vibration, we do not believe a nitric oxide mediated mechanism is involved; rather, a mechanical dislodgement or enhanced lymphatic elimination of gas nuclei is hypothesized. PMID- 20027853 TI - Personality traits in clinically referred aviators: two clusters related to occupational suitability. AB - BACKGROUND: The Five Factor Model (FFM) of normal personality provides a compelling framework for investigating personality subtypes in large military populations. The FFM was used to determine whether a sample of clinically referred military aviators exhibited commonly occurring personality clusters. METHODS: The Revised NEO Personality Inventory (NEO-PI-R) was used to evaluate 956 clinically referred U.S. Naval aviators and flight officers on the domains of neuroticism (N), extroversion (E), openness (O), agreeableness (A), and conscientiousness (C). The scores were subjected to model-based cluster analysis and emergent clusters were compared with respect to their scores and clinical outcomes. RESULTS: A two-cluster model provided the best fit to the data. MANOVA indicated significant differences between the two cluster groups with respect to four of the five factors (N, E, A, and C). The greatest differences were for N and E, with the smaller group (Group 1, N = 291) being significantly more neurotic and less extroverted than Group 2 (N = 665). Cluster membership had more specificity (0.74) than sensitivity (0.59) for predicting adverse clinical outcome, with the probability of an adverse clinical outcome increasing from 0.14 to 0.28 for those in the neurotic and introverted Group 1. CONCLUSIONS: Elevated neuroticism and depressed extraversion were the defining traits of the personality cluster less suited for aviation duty (Group 1). Results support assessment of neuroticism and extroversion during clinical mental health evaluations related to military aviation duty. PMID- 20027854 TI - Organizational pressures and mitigating strategies in small commercial aviation: findings from Alaska. AB - INTRODUCTION: Recent attention has focused on the way in which organizational factors can erode safety in aviation, particularly in regions that have a high accident rate, such as Alaska. The present study builds on this work by examining the direct and indirect pressures that can be exerted on pilots by Alaskan operators. In addition, the paper examines ways in which organizations and individuals manage the effects of pressure. METHOD: Using the critical incident method to uncover situations where the pilot's skills had been challenged, 28 pilots who flew in Alaska were interviewed. A bottom-up qualitative analysis revealed a range of organizational pressures and mitigating strategies. RESULTS: Pilots in Alaska encountered both implicit and explicit norms and expectations to fly in marginal conditions. Pressure also arose from pilots' awareness of the need for their company to make money and from perceived job competition. Some Alaskan operators were able to mitigate the effects of pressure on their pilots and some pilots reported mitigating pressure to fly by managing their employer's expectations and re-emphasizing safety. DISCUSSION: Organizational factors were found to be an important source of pressure for pilots and are likely to contribute to the high accident rate in Alaska. Balancing the competing demands of safety and productivity may be extremely difficult for many small operators, which places a heavy reliance on the decision making of individuals. Both the subtle pressures on individual pilots and strategies for mitigating those pressures are, therefore, extremely important to safety and productivity in small scale commercial aviation. PMID- 20027855 TI - Acute otitic barotrauma during hypobaric chamber training: prevalence and prevention. AB - INTRODUCTION: Barotitis media is known to be an adverse effect of altitude changes, but few studies have investigated this condition with respect to hypobaric chamber training and the resulting estimations of prevalence vary. METHODS: In order to assess the prevalence of hypobaric chamber-related barotitis and evaluate a method of prevention, 335 healthy military pilots undergoing high altitude training were subject to clinical examination and tympanometry before entering the chamber. In order to minimize the risk of barotrauma, only subjects with normal preflight findings were cleared for altitude exposure. Barotitis media was diagnosed on the basis of ear pain and clinical findings according to Teed's classification. RESULTS: Barotitis occurred in eight subjects; seven cases were monolateral and one bilateral, prevalence was 2.4%. CONCLUSION: The prevalence of barotitis after hypobaric chamber training is low in our study, suggesting that a pre-chamber medical check including clinical examination and tympanometry could be effective in identifying subjects at risk. PMID- 20027856 TI - Ultrasound evaluation of sinus fluid levels in swine during microgravity conditions. AB - BACKGROUND: Acute rhinosinusitis is a common problem that could occur in space secondary to absence of gravity-dependent drainage or odontogenic or external sources of infection. The purpose of this study was to determine the efficacy of ultrasound to determine sinus fluid distribution levels in swine and to assess the accuracy of ultrasound in the animal during normal and microgravity conditions. METHODS: Anesthetized swine had a catheter placed through a frontal bone window to allow aliquots of a viscous solution to be injected at 1 G (N = 4) or during brief microgravity parabolic flights (N = 4). Ultrasound examinations were performed with a high frequency probe during baseline and fluid-induced conditions. RESULTS: There was a consistent air-fluid level interface seen on ultrasound examination with the injection of 1 ml of fluid during 1-G conditions. Microgravity conditions caused the rapid (< 10 s) dissolution of the air-fluid level associated with dispersion of the fluid to the walIs of the sinus cavity in a uniform fashion. The air-fluid interface was recreated with return to 1 G. CONCLUSIONS: Ultrasound is a reliable diagnostic test for assessing fluid levels; these experiments demonstrate the technique can be used during microgravity conditions with attention to altered fluid behavior in the absence of gravity. PMID- 20027857 TI - Hypoxic syncope in a competitive breath-hold diver with elevation of the brain damage marker S100B. AB - INTRODUCTION: Competitive breath-hold divers can accomplish previously unbelievable performances; e.g., the current world record for apnea during rest ("static apnea") is 11 min 35 s. However, whether such performances are associated with a risk for hypoxic brain damage has not been established. CASE REPORT: A breath-hold diver's competitive performance resulted in a loss of consciousness, after which he was subjected to a medical examination by the event physician. Blood samples were collected for analysis of the brain damage marker S100B in serum. The S100B in serum was 0.100 microg x L(-1) in the blood sample collected 15 min after the loss of consciousness. At 1 and 5 d after the incident it was 0.097 microg x L(-1) and 0.045 microg x L(-1) respectively. DISCUSSION: The elevated level of S100B, close to the upper reference limit (0.105 microg x L(-1)) indicates that the incident affected the integrity of the central nervous system. Even though this case does not establish that hypoxic brain damage is an inherent risk with loss of consciousness in competitive breathhold diving, the observation raises concerns. We suggest that it should be considered that repetitive exposures to prolonged apneas leading to severe hypoxia may be associated with negative long-term effects. PMID- 20027858 TI - A tribute to the U.S. Air Force Medical Service on its 60th anniversary. AB - Chief of Staff of the U.S. Air Force, Gen. Norton Schwartz, delivered the following remarks, adapted for print, during a dinner celebrating the 60th anniversary of the USAF Medical Service on June 25, 2009, in San Antonio, TX. In commemorating the occasion, he remarked on the history of the USAF Medical Service and went on to commend those who serve today: "Courageous and dedicated medical professionals--who help pull the wounded off the battlefield, treat them en route, and provide sophisticated life-saving treatment and rehabilitation- underpin many incredible stories of survival by critically wounded personnel." He concluded by thanking those in the USAF Medical Service for their dedication. PMID- 20027859 TI - Regarding cataracts in airline pilots. PMID- 20027860 TI - Fatigue risk management in flight crew scheduling. PMID- 20027861 TI - You're the flight surgeon. Allergic contact dermatitis. PMID- 20027862 TI - Classics in space medicine. Possible methods of producing the gravity-free state for medical research. PMID- 20027863 TI - Short nucleotide sequences signal spliceosomal binding in nucleic acids. AB - We have explored the region around the splice sites of the human intron and exons from the exon-intron database (EID) and located a number of short 6-nucleotide and 7-nucleotide sequences that are relatively common in the regions. These short sequences, we expect play an important role in the selection of the appropriate splicing process. We propose that the external signals via short recognition sequences play the deterministic role in the actual splicing process. We have obtained 50 such sequences each from the exon and intron from the beginning and from the ending and noted a number of common features. PMID- 20027864 TI - Purification and characterization of mycoferritin from Aspergillus flavus MTCC 873. AB - The fungus Aspergillus flavus MTCC 873, a non-toxigenic isolate demonstrated its capability to synthesize mycoferritin (MF) upon induction with iron in yeast extract sucrose (YES) medium. The molecular mass, yield, iron and carbohydrate contents of the MF were 440 kDa, 0.015 mg/g of wet mycelia, 0.8 and 30.4%, respectively. Native gel-electrophoresis revealed a band corresponding to dimeric form of equine spleen ferritin (ESF). Subunit analysis by SDS-PAGE revealed a single protein band with an apparent molecular mass of 24 kDa, suggesting similar sized subunits in the structure of apoferritin shell. Immunological cross reactivity was observed with the anti-fish liver ferritin. Transmission electron microscopy (TEM) revealed an apparent particle size of 100 A. N-terminal amino acid sequence of MF revealed a sequence of SLPLQDYA, which showed identities with other eukaryotic ferritin sequences. The spectral characteristics (UV/VIS, fluorescence and circular dichroic spectra) were similar to ESF. The fungus, unlike A. parasilicus 255 (non-toxigenic) was incapable of producing allatoxins, when grown in YES media. PMID- 20027865 TI - Efficient and rapid purification of lentil alpha-galactosidase by affinity precipitation with alginate. AB - Alpha-Galactosidase (alpha-D-galactoside galactohydrolase, EC 3.2.1.22) was purified (26-fold) from the germinating seeds of lentil (Lens culinaris) by affinity precipitation with alginate. The purified enzyme gave a single band corresponding to molecular mass of 40 kDa on SDS-PAGE. The optimum temperature and pH of the enzyme were determined as 40 degrees C and 5.5, respectively. The enzyme was very stable at a temperature range of 4-65 degrees C and at a pH range of 4-7. The values of kinetic constants Km and Vmax using p-nitrophenyl-alpha-D galactopyranoside (PNPG) as substrate were 0.191 mM and 0.73 U, respectively. Results suggest that affinity precipitation is an attractive process for the purification of alpha-galactosidase. PMID- 20027866 TI - Protective role of Cassia auriculata leaf extract on hyperglycemia-induced oxidative stress and its safety evaluation. AB - Cassia auriculata L. (Caesalpiniaceae) is widely used from the ancient period to treat diabetes mellitus. In the present study, the antioxidant activity of C. auriculata aqueous leaf extract (CLEt) was evaluated in streptozotocin-induced mild diabetic (MD) and severe diabetic (SD) rats. A short-term toxicity assessment was also conducted in healthy rats to examine toxic effects of the extract. Oral administration of CLEt to MD and SD rats (100, 200 and 400 mg/kg body weight per day for a period of 21 days) produced significant fall in fasting blood glucose (FBG) in a dose-dependent manner. Treatment with the extract (400 mg/kg) showed significant reduction in serum levels of thiobarbituric acid reactive substances (TBARS) and oxidized low-density lipoprotein (OxLDL) in both MD and SD rats. The antioxidant defense system was also found to be improved in CLEt-treated (400 mg/kg) MD and SD rats, as revealed by significant increase in activities of erythrocyte's antioxidant enzymes, i.e., superoxide dismutase (SOD) and catalase (CAT) with a concomitant elevation in erythrocyte's reduced glutathione (GSH) content. Moreover, there were no toxic signs in rats treated with high doses of the extract (1000 and 2000 mg/kg body weight per day for 21 days). Blood glucose, hepatic and renal function parameters in these rats were found within normal limits. Phytochemical screening of CLEt revealed the presence of alkaloids, flavonoids, saponins, tannins and cardiac glycosides with antihyperglycemic and antioxidant properties. This study suggests that CLEt possesses potent antioxidant activity along with antihyperglycemic potential, hence protective against diabetic complications. PMID- 20027867 TI - Effect of gallic acid on alkaline phosphatase and peptidase activities in rat intestine. AB - Gallic acid is a normal constituent of many edible foods, thus directly interacts with epithelial tissue in intestine. In the present study, the effect of gallic acid on intestinal alkaline phosphatase (IAP) and peptidase activities in rat intestine was evaluated. Gallic acid (0.27-0.5 mM) inhibited activities of leucine aminopeptidase (LAP) and y-glutamyl transpeptidase (y-GTP) by over 90%, compared to controls in rat intestine. In contrast, 0.1-0.6 mM gallic acid either had no effect or stimulated the activity of IAP in rat intestine. The observed inhibition of peptidases by gallic acid was reversible in nature. Kinetic analysis revealed no change in Vmx of LAP (0.42-0.44 units/mg protein) and gamma GTP (0.22-0.24 units/mg protein), while the values of apparent Km were increased 6-7 fold, exhibiting competitive-type of enzyme inhibition by gallic acid. The values of Ki for LAP and gamma-GTP were 0.037 mM and 0.017 mM, respectively. These observations indicate that gallic acid is a potent inhibitor of brush border peptidases, and thus may interfere in the digestion and absorption of proteins in the intestine. PMID- 20027868 TI - Oxidation of oxymyoglobin by poplar plastocyanins a and b. AB - Oxidation of oxymyoglobin [MbO2(Fe2+)] by isoplastocyanins a (PCa) and b (PCb) was experimentally investigated and the corresponding redox reaction was modeled using the physicochemical parameters of the isoforms to study the effect of the dimorphism. The kinetic curve of oxidation of MbO2 (Fe2+) by oxidized PCa [PCa(Cu2+)] and PCb [PCb(Cu2+)] and the pH-dependence of the rate constant kI were determined. In the range of pH 4.8-9.0, PCb reacts with higher k1, compared with PCa. For example, at pH 7.0, k1(PCb) = 4 x 10(2) M(-1)s(-1), whereas k1(PCa)= 2 x 10(2) M(-1)S(-1). The observed values of deltaE(0) for the reaction pairs Mb-PCa and Mb-PCb were -304 mV and -319 mV, respectively. The effect of the ionic strength (mu) on the rate of the electron transfer was also studied. It was found that: (i) the net charge Z1 of PCa and PCb fully corresponds to that calculated by their primary structures and Z2 of Mb corresponds to that calculated by its titration curve; (ii) the In k as function of mean square of mu was similar for both PCa and PCb; (iii) the curve of the reaction PCb <----(e( 1)) Mb (pH 7.0) wasshifted towards higher values of k, in agreement with the larger net negative charge of PCb; and (iv) the character of the electrostatic interactions remained unchanged by a replacement of PCa by PCb and by the change of pH from 7.0 to 4.8. PMID- 20027869 TI - Kinetics of oxidation of adenosine by tert-butoxyl radicals: protection and repair by chlorogenic acid. AB - The rates of oxidation of adenosine and chlorogenic acid by tert-butoxyl radicals (t-BuO) were studied by measuring the absorbance of adenosine at 260 nm and chlorogenic acid at 328 nm spectrophotometrically. t-BuO radicals were generated by the photolysis of tert-butyl hydroperoxide (t-BuOOH) in presence of tert-butyl alcohol to scavenge OH radicals. The rates and the quantum yields (phi) of oxidation of chlorogenic acid by t-BuO radicals were determined in the absence and presence of varying concentrations of adenosine. An increase in the concentration of adenosine was found to decrease the rate of oxidation of chlorogenic acid, suggesting that adenosine and chlorogenic acid competed for t BuO radicals. From competition kinetics, the rate constant of chlorogenic acid reaction with t-BuO was calculated to be 3.20 x 10(9) dm3 mol(-1) s(-1). The quantum yields (phi expt) were calculated from the experimentally determined rates of oxidation of chlorogenic acid under different experimental conditions. Assuming that chlorogenic acid acts as a scavenger of t-BuO radicals only, the quantum yields (phi cal) were theoretically calculated. phi expt and phi cal values suggested that chlorogenic acid not only protected adenosine from t-BuO radicals, but also repaired adenosine radicals, formed by the reaction of adenosine with t-BuO radicals. PMID- 20027870 TI - Analysis of comparative efficiencies of different transformation methods of E. coli using two common plasmid vectors. AB - The efficiencies of different transformation methods of E. coli DH5Qalpha train, induced by several cations like Mg2+, Mn2+ Rb+ and especially Ca2+, with or without polyethylene glycol (PEG) and dimethyl sulfoxide (DMSO) were compared using the two commonly used plasmid vectors pCAMBIA1201 and pBI121. The widely used calcium chloride (CaCl2) method appeared to be the most efficient procedure, while rubidium chloride (RbCl) method was the least effective. The improvements in the classical CaCl2 method were found to further augment the transformation efficiency (TR)E for both the vectors like repeated alternate cycles of heat shock, followed by immediate cold, at least up to the third cycle; replacement of the heat shock step by a single microwave pulse and even more by double microwave treatment and administration of combined heat shock-microwave treatments. The pre treatment of CaCl2-competent cells with 5% (v/v) ethanol, accompanied by single heat shock also triggered the (TR)E, which was further enhanced, when combined heat shock-microwave was applied. The minor alterations or improved approaches in CaCl2 method suggested in the present study may thus find use in more efficient E. coli transformation. PMID- 20027871 TI - Partial purification and some properties of alpha-amylase from Bacillus subtilis KIBGE-HAS. AB - An extracellular alpha-amylase from Bacillus subtilis KIBGE-HAS was partially purified by ultrafiltration and ammonium sulphate precipitation with 19.2-fold purification and specific activity of 4195 U/mg. The enzyme showed relatively high thermostability and retained 62% of its activity when kept at 70 degrees C for 15 min. alpha-Amylase was highly stable at -18 degrees C and loss of activity was very low during stability study. Metal ions like Mn2+ Ca2+, Co2+, K+, Mg2+, and Fe3+ activated the enzyme, while Hg2+ Ba2+, Cu2+, Na+ and Al3+ strongly inhibited the activity. The a-amylase was highly stable in various surfactants and detergents. In the presence of surfactants such as SDS and Triton X-100 the enzyme activity was found 2.9 and 1.8-fold higher respectively than control. The non-ionic detergents (Tween 20 and Tween 80) exhibited slightly inhibitory effect on the enzyme activity. PMID- 20027872 TI - Effect of mercury ion on the stability of the lipid-protein complex of isolated chloroplasts. AB - Mercury is known to interact with different parts of living systems causing serious biochemical and physiological disorder. In order to know the effect of mercury (Hg2+) ion on chloroplasts, the cell free organelle are incubated in an isotonic buffer medium in presence of mercury ion. The metal ion is found to induce membrane lipid peroxidation, loss of photosynthetic pigments and degradation of proteins. Such degradation brings about a drastic modification of lipid-protein organization of chloroplasts as reflected from a blue shift of absorption peaks and lowering of chlorophyll-a fluorescence intensity. The detrimental effect of Hg2+ ion has been explained in terms of direct binding with lipid-protein complex of photosynthetic membrane. Such a binding of metal ion exposes the lipid-protein complex for an easier entry and attack of reactive oxygen species (ROS) generated during incubation of chloroplasts in light and dark, thereby resulting in higher disorganization, which is evident from cation- induced changes in absorption and emission characteristics of the organelle. PMID- 20027873 TI - Learning from the physician integration mistakes of the past. PMID- 20027874 TI - Last throes of healthcare reform 1.0. AB - After focusing on healthcare reform for most of 2009, the House and the Senate are in the final stages of reconciling the differences among their various bills both within and between the two houses of Congress. PMID- 20027875 TI - Finding gold in the supply chain. AB - To optimize supply chain performance and achieve savings that enhance the bottom line, hospitals should: Generate purchase orders for every item purchased. Ensure the organization is protected from undue vendor influence. Keep track of where supply chain dollars are going. Review contracts regularly to ensure competitiveness. Have a contract for each product category. Buy only what the organization is sure to use. Get rid of excess inventory. Develop a strategic plan for continued supply chain savings. PMID- 20027876 TI - Suzanne Delbanco: using technology to improve quality and patient safety. PMID- 20027877 TI - Multiple paths to integrated health care. AB - U.S. health systems appear to be making considerable progress in becoming more integrated and in defining their own, unique approaches to integration. Healthcare reform is likely to lead to a rapid acceleration of this trend. The most critical consideration in the integration strategy for any health system is to attend to the role physicians will play in the more integrated system; and the role of physician leadership, in particular, is of quintessential importance. PMID- 20027878 TI - Doing the math on physician employment. AB - When assessing the costs and benefits of a physician employment strategy, it's important to consider not only the direct costs and benefits of the strategy, but also its indirect benefits, such as increased revenue as a result of eliminating admission-splitting by employed physicians. The indirect risks of physician employment, such as eliminating independent physicians, can be minimized by adjusting timing to match the market's physician employment stage. The strategy will work if the combined direct and indirect benefits are significant enough to outweigh the direct costs. PMID- 20027879 TI - The integration payoff: 3 examples. AB - Physician integration strategies that can lead to enhanced clinical and financial outcomes include: Adoption of clinical IT strategies. Dashboard metrics for measuring the ROI of integration that are almost singularly focused on improving clinical performance. A culture of innovation. PMID- 20027880 TI - The legal outlook for physician-hospital relationships. AB - Legal developments that could affect the relationship between hospitals and physicians include: Enforcement of fraud and abuse laws. Tighter billing rules. Emphasis on value-based purchasing. Renewed interest in alternative delivery models that reward quality, efficiency, and cost savings. PMID- 20027881 TI - Hospital-physician alignment: no decision is a decision. AB - Deliberate discussions on hospital-physician alignment or relationships are critical as a strategy to manage the effect of healthcare reform. There are many competitive threats to be considered in hospital-physician alignment. Hospitals and physicians should take careful steps to determine the structure and needs of each group in development of a high-quality healthcare system. PMID- 20027882 TI - Improving patient flow with a support service call center. AB - Using a call center model for central transport and housekeeping services is key to improving patient flow. Costs for creating a call center environment are incurred for labor and technology. The hospital's financial executive can help the planning team perform the cost-benefit analysis. PMID- 20027883 TI - Understanding the quality of earnings for integrated health systems. AB - Integrated delivery systems could benefit from analyzing quality of earnings to better understand their management plans and methods. Quality-of-earnings analyses can provide understanding of current and future financial performance beyond the bottom line. Following a framework for analysis can help leaders gain a better overall picture of the system's finances. PMID- 20027884 TI - Measuring meaningful use. PMID- 20027885 TI - What would you do? Can a merger create a vital health system? PMID- 20027886 TI - Liquidity meets the new normal. PMID- 20027887 TI - Trends in malpractice costs per adjusted patient day. PMID- 20027888 TI - [What is the interest in cancers in our aged patients?]. PMID- 20027889 TI - [Grief in the aged, nurse are in the first line]. PMID- 20027890 TI - [Main cancers under consideration in the aged]. PMID- 20027891 TI - [Alzheimer disease, 135,00 French persons may be waiting]. PMID- 20027892 TI - [Every year, 80,000 more people pass the age of 80 years]. PMID- 20027893 TI - [Taking care of the aged who have problems with alcohol drinking]. PMID- 20027894 TI - [Coordination of home nursing day and night services]. PMID- 20027895 TI - [Psychology of home nursing for the aged]. PMID- 20027896 TI - [Ethics and the aged. Combining care and laying a wager]. PMID- 20027897 TI - [These nurses who defend the common weal]. PMID- 20027898 TI - [Is it necessary to legislate for institutionalization without consent?]. PMID- 20027899 TI - [What are the ethics of Alzheimer disease?]. PMID- 20027900 TI - [Access of aged persons with Alzheimer disease and cancers to nurses]. PMID- 20027901 TI - [The Leonetti Law and geriatric ethics]. PMID- 20027902 TI - [The Leonetti Law in practice]. PMID- 20027903 TI - [Struggle against abuse, a complex ethic]. PMID- 20027904 TI - [Ethical considerations on care of the patient with Alzheimer disease]. PMID- 20027905 TI - [Refusal of nutrition in a patient with Alzheimer disease]. PMID- 20027906 TI - [In which ethical setting does the everyday nurse find herself?]. PMID- 20027907 TI - [6/6 Eye infections]. PMID- 20027908 TI - [The two (and more) cultures of the "clone". Utopia and fiction in post-war discourses of life sciences]. AB - Since the late 1950s, "two cultures" has become a catch phrase for describing a deep divide between science and literature. When Charles P. Snow, who initiated this discussion, introduced the notion of "two cultures" in a lecture at the University in Cambridge in 1959, he referred to an incompatibility of scientific and literary worldviews in Western Societies. His thesis of two contradicting cultures immediately received a huge variety of different responses from philosophers, scientists, novelists and literary scholars. However, this article argues that this widespread debate was part of a broader post-war discourse on the impact of modern science on society, in which especially the idea of "scientific progress" was at stake. Central to this debate was the question of how scientific and technological progress could affect the notion of the "human" itself. The paper analyses the emerging discourse on cloning against this background. The constitutive role of fiction and imagination in both fields, science and literature, is explored by tracing the scientific, utopian and literary cultures in which figures of human clones have taken different shapes since the 1960s. At that time, scientists developed utopian views in which the "clone" became a metaphor for future possibilities of transcending and reshaping the human nature. Science fiction writers reacted to this by portraying the human clone as an individual and by depicting human clone figures in a psychological way PMID- 20027909 TI - [Psychophysical parallelism. On a discursive figure in the field of scientific changes in the late 19th century]. AB - The article traces the rise and fall of "psychophysical parallelism" - which was the most advanced scientific formulation of the mind / body relationship in the second half of the 19th century - through an interdisciplinary and broad geographical spectrum. It sheds light on the extremely different positions that rallied round this discursive figure, ranging from Fechner, Hering, Mach, Wundt, Bain, Hughlings Jackson, and Taine to Freud and Saussure. The article develops the thesis that the psychophysical parallelism functioned as a 'hot zone' within and a symptom of the changes in the order of sciences at that time. Against that background, the criticism of the psychophysical parallelism which became prominent around 1900 (Stumpf, Busse, Bergson, Mauthner et. al.) indicates the cooling of this 'hot zone' and the establishment of a new order within the scientific disciplines. The article pays particular attention to the position of this figure in contemporaneous language theories. Its basic assumption is that the relationship between the body and the psyche is itself constituted by language. PMID- 20027910 TI - The scientific claims of British child guidance, 1918-45. AB - This article examines the British child guidance movement's claim to scientific status and what it sought to gain by the wider acceptance of such a claim. The period covered is from the movement's origins in the 1920s to the end of the Second World War, by which point it had been incorporated into the welfare state. This was also an era when science commanded high intellectual and cultural status. Child guidance was a form of psychiatric medicine that addressed the emotional and psychological difficulties that any child might experience. It thus saw itself as a form of preventive medicine and as a component of the international movement for mental hygiene. Child guidance was organized around the clinic and employed the knowledge and skills of three distinct professions: psychiatrists, psychologists and psychiatric social workers. Its claim to scientific status was underpinned by the movement's clinical and organizational approach and in turn derived from developments in the laboratory sciences and in academic medicine. There were, however, those even within the movement itself who challenged child guidance's purported scientific status. Such objections notwithstanding, it is suggested here that at least in its own terms the claim was justified, particularly because of the type of psychiatric approach which child guidance employed, based as it was on a form of medical holism. PMID- 20027911 TI - Between less eligibility and the NHS: the changing place of poor law hospitals in England and Wales, 1929-39. AB - In 1929, the Local Government Act broke up the apparatus of the Poor Law Guardians and Unions, and transferred responsibility for the care of the poor to local councils. In theory, the period between the passing of the Act and the formation of the National Health Service witnessed a large-scale reclassification of the sick poor as patients rather than paupers. In reality, as this investigation of contemporary judgements of hospital quality and bed and staff numbers in English and Welsh county boroughs shows, the national picture was very varied at the local level. Local and sometimes regional traditions of care, finance and council priorities had a large influence on the ongoing development of a unified medical service which included the poor. In the best case scenario, hospitals were classified by patient type, and the principle of 'less eligibility' was discarded. Elsewhere, economic status continued to direct medical treatment, but in almost all cases, the chronic and elderly poor were more likely to remain in low-quality and unmodernized buildings than the acutely sick. The investigation highlights the disjuncture between the changed vision for the sick poor and its patchy enforcement on the ground. PMID- 20027912 TI - Taking care of oneself in the twentieth century: a postcard story. PMID- 20027913 TI - A chronological outline of the history of the Poudre des Chartreux, or Carthusian Powder, or Mineral Kermes. PMID- 20027914 TI - From history of pharmacy to pharmaceutical history. PMID- 20027915 TI - From mercury to miracle drugs: syphilis therapy over the centuries. PMID- 20027916 TI - The discovery of Trypanosoma cruzi and Chagas disease (1908-1909): tropical medicine in Brazil. AB - This article analyzes the discovery of Chagas disease and the parasite that causes it (Trypanosoma cruzi) by Carlos Chagas in 1908/1909, with a special focus on the scientific and social context in which this occurred. Its inclusion in the international debate on European tropical medicine--especially with researchers from the German school of protozoology--and its connection with discussions on the modernization of the recently established Brazilian Republic are also examined. The discovery of Chagas disease became a decisive aspect in the scientific project that Oswaldo Cruz sought to establish at the institute that bears his name. It was extolled as a symbol of Brazil's scientific ability t produce knowledge in line with the international scientific agenda, while simultaneously being attuned to the specific problems of the country. PMID- 20027917 TI - The reception by French physicians of Chagas' discovery of Trypanosoma cruzi and American trypanosomiasis (1909-1925). AB - As soon as they were published early in 1909, Chagas's articles on Trypanosoma cruzi and American trypanosomiasis became the topic of discussions in France. The description of T. cruzi and Chagas disease was added to parasitology textbooks as early as 1912, and elicited active research, particularly on the part of French parasitologist Emile Brumpt. He contributed towards eluciding the lifecycle of T. cruzi and the different ways it could infect humans. Laboratory research on T. cruzi was interrupted by First World War and was not resumed afterwards on the same scale, although interest in the epidemiology of Chagas disease continued. PMID- 20027918 TI - [The history of Chagas' disease in Argentina: conceptual, institutional, and political evolution]. AB - In the one hundred years since the identification of Chagas disease, major changes have occurred in its scientific conception, institutional recognition, and political weight. From a medical perspective, it was seen as the cause of goiter, next its acute effects were emphasized, and then its effects on cardiac health received greater attention. In similar fashion, sanitary policy first downplayed the disease's importance, then elevated it to the role of a national cause, and gradually relegated it to the bottom of the agenda. The article briefly presents the key points of this historical trajectory in Argentina, exploring the cognitive, political, and institutional underpinnings of the disease as both a social and biological fact. PMID- 20027919 TI - Trypanosoma cruzi, cancer and the Cold War. AB - In the summer of 1946, the international community of cancer researchers was inspired by the announcement that two Soviet scientists, Nina Kliueva and Grigorii Roskin, had discovered anticancer properties in culture extracts made from the South American protozoan, Trypanosoma cruzi, and had produced a preparation--named after its discoverers KR--which showed clear therapeutic effects on cancer patients. Research teams from various countries enthusiastically pursued the promising new line of investigation. The story of the rise and fall of interest in the anticancer properties of T. cruzi in different countries suggests that during the second half of the twentieth century, the Cold War competition between the superpowers played an important role in shaping the research agendas of cancer studies. PMID- 20027920 TI - [The cardiological perspective: Francisco Laranja and research on Chagas' disease. Interview by Rose Goldschmidt, Jaime Benchimol, Marcos Chor Maio]. AB - Francisco Laranja began working in cardiology when this was yet a fledgling specialty in Brazilian medicine. An expert in electrocardiography, he has gained national and international renown for his research conducted in Bambui in the 1940s, which led to the clinical characterization of the cardiac profile of chronic cases of Chagas' disease. In this interview, Laranja talks about the development of the field of cardiology and electrocardiography in Brazil, his work at the Institute of Retirement and Pensions for Industrial Workers, his research in Bambui, and his term as director of the Oswaldo Cruz Institute. PMID- 20027921 TI - [Interpreting Brazil as afflicted by disease and by the spirit of routine: the repercussion of Arthur Neiva and Belisario Penna's medical report (1917-1935)]. AB - The release of a report on the Oswaldo Cruz Institute's 1912 scientific voyage to North and Northeast Brazil, led by physicians Arthur Neiva and Belisario Penna, debate that found its way to the pages of magazines of the letters and sciences. The report used the images of disease, geographic and cultural isolation, illiteracy, poverty, and a vocation for backwardness to portray the people living in interior Brazil. These images of the sertao were extensively criticized in the periodical A Informacao Goiana, published by local doctors who refused to see the interior defined as 'sickly' and 'backwards'. The article analyzes the ways in which the Neiva-Penna report distinguished itself becoming a reference for intellectual controversies surrounding the national question in Brazil. PMID- 20027922 TI - [Carlos Chagas and the debates and controversies surrounding the disease of Brazil (1909-1923)]. AB - The article explores the relation between the debate on Chagas' disease, discovered in 1909, and Brazil's 1916-1920 rural sanitation campaign. It argues that the political planks of the sanitary movement were intimately bound up with the definition and legitimization of this illness as a scientific fact and social issue. Presented as emblematic of rural endemic disease, this 'new tropical ailment' was characterized as 'the disease of Brazil', symbol of a 'sickly country'. The sanitary campaign was in turn a decisive element of the 1919-1923 polemic surrounding the disease. This is an exemplary case of how Brazilian scientists used theories from European tropical medicine to produce original knowledge in the field, basing themselves on meanings specific to the national context of their day. PMID- 20027923 TI - [A Brazilian medical collection: Central Brazil in Arthur Neiva and Belisario Penna's scientific expedition and Julio Paternostro's voyage to Tocantins]. AB - The article addresses the role played within the social imaginary of Brazil by the scientific voyages of physicians in the first half of the twentieth century. Two texts are analyzed: a report by Arthur Neiva and Belisario Penna published in Memorias do Instituto Oswaldo Cruz and another by Julio Paternostro, released in 1945 in Viagem ao Tocantins. The former contributed to singling out pathology as defining mark of national identity during the First Republic (1899-1930), a fact that had repercussions in the following decades, as apparent in Paternostro's book, which at the time of its publication was presented as an indictment of national problems. These portraits of Brazil highlight as attributes of the country not only disease but also the geographic and, primarily, cultural distance separating the coast from the sertao. PMID- 20027924 TI - [The scientific journey taken by Neiva and Penna: a blueprint for studies of diseases from the Brazilian hinterland]. AB - Analyze the report by Neiva and Penna, focusing on the contribution these authors made to the study of one of the endemic diseases encountered throughout their journey, which had been long known under the popular name of mal de engasgo (choking disease). They recorded their observations and detailed descriptions of the patients' symptoms and the association frequently encountered between mal de engasgo and another endemic disease, known as vexame or vexame do coracao, characterized by bouts of palpitations. Neiva and Penna's epidemiological and clinical observations of mal de engasgo were crucial for all the researchers interested in the disease, especially its relationship with Chagas disease. PMID- 20027925 TI - [In the heart of Brazil, a healthy capital--the participation of doctors and sanitarists in the construction of Brasilia (1956-1960)]. AB - Projected as an expression of the daring and modernity of an age, Brasilia could not overlook planning that considered the conditions where it would be located in the interior of Brazil. Constructed in a region historically associated with isolation, poverty and diseases, the new capital required the participation of doctors and sanitarists from the very beginning of construction to ensure healthy conditions. Seeing the opportunity to expand their sphere of action, until then restricted to the interior, doctors from Goias stood out in this process, highlighted by concerns of the profession manifested in the periodical published by its association and their extensive mobility and modern practice, contradicting the common conceptions regarding doctors in the interior. PMID- 20027926 TI - ["Brazil isn't only disease": Juscelino Kubitschek's public health program]. AB - Analyzes the public health program that Juscelino Kubitschek presented while running for president and its dialogue with interpretations that identified Brazil as an ailing country that should be recovered through public medicine. The proposals and goals contained in Kubitschek's healthcare program, as released during his 1955 campaign, serve as the main sources for this analysis. They also were key to establishing this dialogue, which was both prompted and shaped by a national and international context characterized by the post-war idea of development and public health optimism. Within the context of his development project, the article looks at how Kubitschek elaborated the themes of the centrality of rural endemic disease and the abandonment of the rural worker, both dear to the sanitary movement in the 1910s. It also underscores how Kubitschek the candidate endeavored to link health and development within this arena of government policy. PMID- 20027927 TI - [The institutionalization of bacteriology in Japan]. AB - Within the history of public health in Japan, Shibasaburo Kitasato is widely known for introducing bacteriology to Japan in the Meiji. This paper looks into the factors behind the success of the Institute of Infectious Diseases(IID), which Kitasato headed, focusing specifically on Kitasato's strategy, institutional rivalries in the medical world and early development in the field of bacteriology. Kitasato used IID to spread bacteriology through a bacteriology training course and the publication of a professional journal, both modeled on the German system, which Kitasato absorbed during his previous studies in Germany under Robert Koch, who was one of the founders of bacteriological science. At the time, doctors related to the university system and those related to sanitary administration competed for leadership in the medical world in Japan. The latter had pushed for IID to be established, and the outbreak of plague in 1899 gave them the opportunity to successfully lobby the Diet into passing legislation that enlarged enrollment in the bacteriology course and earmarked money specifically to IID. In the late 19th century, the application of knowledge from the nascent field of bacteriology led to major advances in the fight against infectious diseases, which led to rapid accumulation of knowledge about bacteria, in turn. Thus, Kitasato emphasized that bacteriology should be mission-oriented and based in sanitary administrations. Indeed, as sanitary administrations were on the frontline of the battle against infectious diseases and as IID laboratories were connected to the sanitary administrations, IID succeeded in dominating the field of bacteriology. PMID- 20027928 TI - [Aleksandr Lwowicz Blok (1852 - 1909)--character-sketch of the man and scientist]. AB - The article is an attempt of a new outlook first of all upon scientific output of many years' (the years 1878 - 1909) lecturer on Russian State Law in the Imperial University of Warsaw--Aleksandr Lwowicz Blok. Historians of literature, researchers of life and output of the scientist's son, an eminent poet-symbolist Aleksandr A. Blok, to a high degree were able to reconstruct lot of life and vicissitudes of the Warsaw professor. Thanks to the preserved university archival materials it was possible to add many details to Blok's biography, especially concerning his professional problems. They were connected with dramatic events in personal life of the professor. According to relations of the part of his university co-workers and students, the circumstances even brought about the symptoms of the mental illness. Prematurely elderly he was a man deeply miserable, lonely and deprived of understanding by associates. Up till now his scientific output has not been described precisely enough. Not numerous memories of his contemporaries (Szymon Askenazy, Nikolaj Dubrowskij) and even inconspicuous modern researches of the Polish (Janusz Wolinski) brought about, in the author's opinion, too simplified and schematic character-sketch of Blok as a Russificator and nationalist, mediocre scholar and much worse lecturer. A scientific output of Blok is, indeed, inostensible--he published 2 books only- Master's thesis entitled: Wladza panstwowa w spoleczenstwie europejskim. Poglad na polityczna teorie Lorenza Steina i na francuska rzeczywistosc polityczna (1880) [State Authority in European Society. Outlook upon Political Theory of Lorenz Stein and French Political Facts (1880)] and Literatura polityczna w Rosj i o Rosji (1884) [Political Literature in Russia and on Russia (1884)] as an introduction to his course in State Law. Two thirds, so over 20 years of work, he gave to the not accomplished conception of creating a new classification of sciences that together with its assumptions was included in work of Blok's disciple--Jewgienij Spektorski. An attempt of objective, free of emotions look at Blok's output to some extent makes us verify one-sidedly its critical estimate. In Blok's writings one can observe several earlier unnoticed values--author's wide mind horizons, his erudition, interesting but not accomplished research conceptions, competence for analysing difficult issues fringing upon social, political and philosophical thought. PMID- 20027929 TI - [Definition of the history of medicine in the interpretation of Wladyslaw Szumowski]. AB - Wladyslaw Szumowski is said to be one of the most eminent Polish historians of medicine. The present text is an attempt of describing Szumowski's ideas of the essence of the history of medicine, its place among sciences and a role it should play in education of future doctors. The paper presents both the definition and the methodology, which in Szumowski's opinion should result from the philosophy, and particularly from the logic. Szumowski is referring to these research inspirations, scientists and conceptions that had a significant influence on him. At the same time the author of the article is willing to extract an original thought of Szumowski and divide it from adopted ideas of other people's conceptions. PMID- 20027930 TI - [Naturalists, collectors and theoreticians of museology]. AB - The origins of the contemporary collectorship dates from times when the sameness of art and science was commonly accepted. In those days relics of the ancient past and natural individuals of newly discovered lands were presented at the same time. Cosmological character of the collections manifested the tenacity of recognition and representation of the surrounding reality. A great impact on completion of collections of curiosities in Europe had Netherlands, and in the basin of Baltic Sea a remarkable significance was gained by Hanseatic Gdansk. Collections of Jakub Breyn, Jakub Klein and Gotfryd Reyger became famous then. In the same way were imported individuals for Anna Jablonowska that composed one of the most interesting European collections. In course of time merging such a great multiplicity of collections was beyond collectors' power and museum pieces from collections of curiosities were parcelled out. It was a real beginning of specialistic museums. A role of museum for science results from its function of methodical organizing collections that can be used by research workers. However, although the aims of scientific and museum centres are different, they come together on the occasion of museum recognition works when museums' workers borrow essential knowledge and methods from resources of science, and scientists search for useful research materials in museum resources. PMID- 20027932 TI - Do the right thing. To pay for health-care reform, both parties need to get serious about a topic neither one mentions: taxes. PMID- 20027931 TI - The strange career of the public option. How a modest idea started a big fight. PMID- 20027933 TI - Swine flu's unsportsmanlike conduct. Athletes are trying to avoid H1N1. But must friendly gestures lose out? PMID- 20027934 TI - Comparison of skin concentrations following topical versus oral corticosteroid treatment: reconsidering the treatment of common inflammatory dermatoses. AB - BACKGROUND: Topical corticosteroids are often considered to have greater safety and poorer efficacy than oral corticosteroids in treating psoriasis and atopic dermatitis. There are limited data for assessing relative efficacy of topical and systemic corticosteroids, however. The concentration of corticosteroid in skin, adjusted for the relative potency of the active compound, may be a predictor of clinical efficacy and can be estimated for both topical and oral administration. PURPOSE: To analyze the assumption that oral corticosteroid therapy should be more potent than topical therapy by comparing relative corticosteroid concentrations in the skin expected with topical versus systemic administration. METHODS: The estimated skin concentration of prednisone following oral dosing was calculated based on data showing 70-100% bioavailability and an even tissue distribution. Data on the concentration of corticosteroids found in skin after topical application were obtained from the literature. The relative potencies of corticosteroid molecules were then used to compare skin concentrations of corticosteroid following topical versus oral treatment. RESULTS: Data derived from the existing literature demonstrated that hydrocortisone 2.5% ointment, triamcinolone 0.1% ointment, and clobetasol 0.05% foam achieved effective skin concentrations greater than the effective concentration achieved by oral prednisone. Betamethasone 0.1% cream achieved effective concentrations in skin within the range created by oral prednisone. LIMITATIONS: This analysis was limited by the paucity of data regarding cutaneous concentrations of corticosteroids after topical application, and by the differing experimental designs utilized in the available studies. CONCLUSION: Most topical corticosteroids have the potential to achieve greater effective drug levels in the superficial layers of skin than those achieved with standard doses of oral prednisone. The apparently greater efficacy of oral corticosteroid therapy may be attributable, in part, to poor patient compliance with topical therapy. Systemic alterations in immune function following oral, but not topical, corticosteroid use may also play a role. PMID- 20027935 TI - Acne quality of life and patient satisfaction following treatment with tretinoin pump. AB - Patient satisfaction and quality of life are important considerations when assessing products used to treat acne vulgaris, as these factors may affect treatment adherence and subsequent treatment outcomes. The objective of this analysis was to determine patient satisfaction and improvement in quality of life after treatment with tretinoin gel microsphere (TGM) in a pump dispenser. Assessments were made during a phase IV, prospective, 12-week, open-label, community-based trial in which 544 patients who were dissatisfied with their current acne treatments received TGM 0.04% or 0.1% in addition to < or = 2 concurrent non-retinoid acne therapies. At week 12, significant improvement was reported in both patient acne therapy satisfaction and in the overall mean Acne Quality of Life Index scale (P < 0.0001 versus baseline for both measures). The majority of patients (82.3%) rated the pump dispenser as an "excellent" or "very good" means of dispensing medication, and 86.0% rated their overall satisfaction with the pump treatment application as "very satisfied" or "extremely satisfied." The results of this study indicate that the use of TGM in a pump dispenser in patients with acne vulgaris is associated with significant increases in both quality of life and patient satisfaction. PMID- 20027936 TI - Hyaluronics for soft-tissue augmentation: practical considerations and technical recommendations. AB - Hyaluronic acid (HA) fillers are long chains of sugar molecules. Depending on various physical properties, such as chain length and cross-linking, they can have different textures and durations. Injections of hyaluronic acids for soft tissue augmentation is one of the most popular procedures performed in the U.S., Europe, Asia and Canada. With the development of newer HA molecules, it is likely that this trend will continue. Choosing the right HA for a particular patient depends on various factors, including the area to be treated, skin thickness and patients' risk tolerance. Understanding the various molecules, and how they interact, is essential for ensuring optimal patient outcomes. PMID- 20027937 TI - Topical corticosteroids in dermatology. AB - Topical corticosteroids are the most commonly prescribed agents in the treatment of dermatologic conditions. They are used primarily as monotherapy or in combination with other agents for enhanced efficacy. Several stronger preparations are now available since their first introduction. They are also available in various vehicles altering the potency and giving the option of tailoring them for use based on specific anatomic locations, area of involvement, age of the patient, and most importantly, severity of the condition. Several local and systemic side effects have been associated with their inadvertent use. Allergic contact dermatitis to most of the preparations has also been noticed. Judicious use with reinforced patient education lowers such risk for side effects, and can be of great use in treating dermatologic conditions. PMID- 20027938 TI - Efficacy of a lipid-based barrier repair formulation in moderate-to-severe pediatric atopic dermatitis. AB - OBJECTIVES: The authors assessed the efficacy of a ceramide-dominant, triple lipid barrier repair formulation (EpiCeram), which designed to correct the lipid biochemical abnormalities in atopic dermatitis (AD) in comparison to fluticasone propionate cream. METHODS: In a five-center, investigator-blinded, randomized trial, EpiCeram was compared to fluticasone (Cutivate) cream in 121 patients with moderate-to-severe AD. Primary outcome measures were: 1) reduction in disease severity, assessed as SCORAD (Severity Scoring for Atopic Dermatitis) scores; 2) improvement in pruritus; and 3) improvements in sleep habits. RESULTS: EpiCeram reduced clinical disease severity, decreased pruritus and improved sleep habits both 14 and 28 days after initiation of therapy. Although the fluticasone-treated group showed significantly greater improvement at 14 days, SCORAD, pruritus and sleep habit scores for EpiCeram did not differ significantly from the fluticasone treated group by 28 days. CONCLUSION: The ceramide-dominant, physiological-lipid based formulation could represent an effective stand-alone or ancillary therapy for many pediatric patients with AD. PMID- 20027939 TI - Three-dimensional imaging techniques in the assessment of facial volume augmentation. AB - Demand for facial volume augmentation procedures is increasing, along with a growing appreciation for the importance of objective, quantitative measures of treatment success. This article reviews currently available three-dimensional (3D) imaging techniques and discusses their practical applications for cosmetic dermatology. X-ray-based imaging techniques can be costly, time-consuming and expose patients to the potential health risks of ionizing radiation. Conventional photographic techniques vary widely in quality. Standardized equipment, with controlled lighting, short exposure times, and high-resolution digital formats, provides high-quality, two-dimensional photographic images allowing accurate assessment of changes in facial features. These images can be reconstructed into 3D surfaces using stereophotogrammetric techniques, offering a major advance in measurement of changes. Although no system is ideal for use in routine clinical practice, stereophotogrammetry-based optical profilers provide acceptable surface quality and image overlays suitable for measurement and visual inspection. For research purposes, fringe projectors yield acceptable image overlays for quantitation of facial volume changes. PMID- 20027940 TI - A novel approach to treatment of the aging hand with Radiesse. AB - The ideal dermal filler for treatment of the aging had needs to have inherent bulk and longevity of effect. The filler calcium hydroxylapatite (CaHA) provides a texture that lends itself to injection and then to subsequent massage, without losing control of where the product is placed. In the recent past, delivery of CaHA to the hands has been accomplished by mixing lidocaine with the dermal filler prior to injection into the dorsum of the hand. The author suggests that an alternative way to deliver CaHA is to administer a large bolus of lidocaine immediately prior to injection of the dermal filler itself, followed by firm massage to disseminate the CaHA microspheres and carrier gel into the areas around and above the tendons and veins of the hand. PMID- 20027941 TI - Case-based experience with the simultaneous use of a fixed topical antibiotic/benzoyl peroxide combination and a topical retinoid in the optimization of acne management. AB - Treatment options for acne vulgaris have expanded considerably in the past decade. The main goals of treatment continue to be reducing acne lesions while maintaining patient satisfaction and adherence to treatment. As dermatologists, the art of treatment is to develop, fine-tune, and utilize combinations of agents to increase compliance, thereby optimizing patient outcomes. In acne management, one strategy involves concomitant use of topical retinoids with benzoyl peroxide or a fixed combination of topical antibiotic/benzoyl peroxide. This strategy requires application of one product in the morning and one in the evening due to concerns of benzoyl peroxide-induced degradation of retinoid activity. Presented are two cases in which a topical retinoid and a topical fixed combination of clindamycin/benzoyl peroxide were used concomitantly in patients with mild-to moderate acne. Research-based and practical rationale on the simultaneous use of newer-generation retinoids and benzoyl peroxide-based products, without concern of retinoid degradation, is discussed. PMID- 20027942 TI - Black tongue secondary to bismuth subsalicylate: case report and review of exogenous causes of macular lingual pigmentation. AB - Macular pigmentation of the tongue can be acquired following exposure to exogenous agents. Black lingual hyperpigmentation was observed during the full body skin examination of a man with a history of recurrent metastatic malignant melanoma. His tongue spontaneously returned to its normal pink color later that day. Bismuth subsalicylate (Pepto-Bismol) was suspected as the pigment-inducing agent; subsequently, re-challange with the antacid confirmed it to be the cause of his acquired, albeit transient, black tongue. The ingestion of medications, including other antacids, analgesics, antidepressants, antihypertensives and several antimicrobials has been associated with the development of acquired macular lingual pigmentation. In addition, hyperpigmentation of the tongue has been observed following the deposition of amalgam and the injection of local anesthesia or doxorubicin or interferon alpha and ribavirin. Also, inhalation of heroin and methaqualone vapors or tobacco has resulted in lingual hyperpigmentation. All of the patients with acquired macular lingual hyperpigmentation had tongues with a smooth surface without enlargement of the filiform papillae. Many of the individuals with hyperpigmented tongue had either black or dark skin color. The onset of tongue pigmentation varied from less than one day to several years after initial exposure to the associated exogenous agent. The color of the tongue usually returned to normal after the pigment inducing agent was discontinued. PMID- 20027943 TI - Adalimumab in the management of palmoplantar psoriasis. AB - Palmoplantar pustular psoriasis (PPP) is an uncommon form of chronic psoriasis. Characterized by sterile, intra-epidermal pustules located on the palms and soles, it is highly resistant to treatment. A patient presented with palmar inflammation and throbbing joint pain in his hands, as well as erythematous, pustular, and micaceous scaling skin on his right foot, legs, elbows and hands. Approximately 4% of his body surface area was involved, and he was diagnosed with PPP after skin biopsy. After conventional therapy failed, the patient underwent treatment with adalimumab and the majority of his symptoms resolved after 16 weeks of therapy. Adalimumab may be effective for the treatment of PPP. Adalimumab, a fully human, immunoglobulin G1 monoclonal antibody that binds to tumor necrosis factor, has been approved for the treatment of moderate-to-severe psoriasis, in the U.S., Europe and elsewhere. PMID- 20027944 TI - Dissolving collagen fillers: enzymatic degradation of some problematic filler circumstances may now include collagens. AB - The market for non-permanent fillers has grown dramatically over the past years. Even though hyaluronic acid fillers and collagen are forgiving, complications, albeit rare, still occur. Up to now, a necessary element of any dermatologic surgeons' office is a hyaluronidase in case one of these complications should occur. However, these agents will only work on hyaluronic acid fillers leaving any problem with collagen products to be resolved by other means. Later this year the first collagenase could be approved for usage in specific medical conditions; at that time it will, perhaps, complement its hyaluronidase counterpart. PMID- 20027945 TI - Cutaneous T-cell lymphoma: a review of current therapies and the future therapeutic implications of chemokine biology. PMID- 20027947 TI - [Family changes due to the hospitalization of a child in a pediatric intensive care unit]. AB - The study aimed at understanding the changes that occurs in a family after the hospitalization of a child in a Pediatric Intensive Care Unit (PICU). It was a descriptive study, with a qualitative approach. Data were collected in the period of March to June 2007, through semi-structured interviews carried out with 21 parents of children admitted in a PICU of a University Hospital located in the northwest area of Parana State, Brazil. The results showed the family, even when facing the hospitalization and the risk of death of a family member, as well as the separation of other family members from the mother who is keeping company to the sick child, tries to establish means to maintain united and to follow its routine. The study revealed the importance of the support to the relatives of a hospitalized child, as an essential part of the caring process, thus contributing to turn it indubitably humanized. PMID- 20027946 TI - [Abortion in adolescence: who committed it in Maceio, Alagoas, Brazil]. AB - This quantitative and cross study aims at establishing a profile of female teenagers, from 12 to 19 years old, that committed abortion in a representative random sample of 2592 adolescents in Maceio, Alagoas, Brazil, in 2005. There were found 149 abortions, where most of them (81.9%) did not have a job, (85%) lived with the parents, (85.2%) were single, (69.8%) studied in a public school and (70.5%) were in high school. The majority had one abortion 93%, but it stands out that 8.5% of them aborted twice and 2.6% four times, showing this data the inadequate attention to teenagers reproductive health. In order to have an abortion, 63.8% of them had support, 83.9% did not have physical complications and 89.3% did not need hospitalization. The conclusion is that the teenagers are getting pregnant and still attending school, looking for abortion as a solution for a not planned pregnancy and that the lack of need of hospitalization post abortion contributes to the statics lack. PMID- 20027948 TI - [Seeds of palliative care: nurses' discourse order]. AB - Palliative care is becoming a professional knowledge and performance field by means of the proposal of another "regimen of truths". Such fact can be observed in the broadening of palliative care services and in the increase of publications on the subject. This study aims at learning the discourses on palliative care that the nurses have conveyed in the nursing publications. For such purpose we approached the Cultural Studies, especially those inspired on Michel Foucault in order to analyze articles published in two national nursing periodicals from 1999 to 2007. Among the statements, we point out those that see the hospital as a site where the patient is divested of his individuality and identity and those that mention such care as capable of providing the family and the patient with the best quality of life possible during the death process. PMID- 20027949 TI - [Evaluation of the self-esteem of bearers of HIV/AIDS of Alfenas city, Minas Gerais, Brazil]. AB - This study aimed at identifying the socioeconomic characteristics and at evaluating the self-esteem of the adult users with positive serology anti-HIV and attacked by AIDS, assisted in the Centro de Testagem e Aconselhamento (CTA) of the Alfenas, Minas Gerais, Brazil. It is a study accomplished among April to August 2008 with the technique of semi-structured interview. The age group prevailed among 30-39 years (37.78%), feminine gender (53.3%), white color (48.9%), single (37.8%), low education level (42.2%), that they acquired the virus through sexual relationship (88.9%) and with medium score of self-esteem (62.2%). In relation to the relatives, 24.44% of the interviewees had at least a positive serum in the family. The epidemic identification profile of the bearers of HIV/AIDS in CTA of Alfenas it comes to evidence the need of larger social mobilization and of inter-sectorial integration in the implementation of actions that can act facing this grievance. PMID- 20027950 TI - [Family evaluation: social support network in mental health care]. AB - This study aims at identifying the social support network of a user and his family in a Psychosocial Care Center (CAPS) follow-up. Such study is a subproject of the research named Evaluation of Psychosocial Care Centers, from the southern region of Brazil, at a CAPS of Porto Alegre, Rio Grande do Sul, Brazil. The subjects were a user and a relative of him. It is a qualitative research of case study type that makes use of the Calgary Model of Family Evaluation. The collecting of data provided the construction of the genogram and of the eco-map. The analysis focused on structural, functional and developmental evaluations. The user's bonds and his relationships with the micro and macro family space were identified by showing the social support networks and the functions that they perform in the user's daily life. The social support networks are an important therapeutic resource because they provide the possibility of offering a treatment addressed to the life of the subject. They also take in count his culture and they broaden care actions as well. PMID- 20027951 TI - [The care experience in the context of home health care]. AB - This study aimed at describing and analyzing family carers' experiences in taking care of chronic patients at home. It is a qualitative research, developed in Service of Home Health Care of Santa Maria's University Hospital (SIDHUSM), Rio Grande do Sul, Brazil, to which 15 family carers contributed. In order to produce data, one made use of Creative-Sensitive Method as well as the dynamics of creativity and sensitivity: Life Line, Body-Knowledge and Almanac. Data were analyzed according to the conceptual premises of discourse analysis. Thus, there were developed the following themes: the care as an inherent experience to human beings; the experience of taking care in a disease situation; the learning with the experience of being a carer. The results pointed to the necessity of valuing the care activities developed by home carers at patients' home, and they also highlighted the home care as a modality of humanizing and innovating attention which aims to invert health professionals' logic of action. PMID- 20027952 TI - [Institutional characteristics that interfere in the leadership of nurses]. AB - The development of this study aimed at knowing the institutional characteristics that interfere with leadership of the nurse. This is a qualitative survey of descriptive and exploratory type, held in a large hospital located in the southern Rio Grande do Sul, Brazil, during the months of February and March 2008, in which participated 11 nurses. For the collection of data, we used semistructured interview. It was opted for the analysis of content as a technique for data processing in which emerged a category concerning institutional character that consists of six subcategories: remuneration, material resources, autonomy and institutional support, rotation, devaluation of the profession, and lack of incentive to develop leaders. It was identified that the little incentive to the improvement of the nurses front to the leadership is one institutional problem that can be minimized through the permanent education and the creation and implementation of programs of development for leaders in nursing. PMID- 20027953 TI - [Nursing consultation in a health vigilance program: the Pra-Nene experience]. AB - This study focus on describing the opinion of the parents of children belonging to the "For-Baby Program", about the educational activities in health performed by the nurses in a nursing consultation at a Health Basic Unit of Porto Alegre, Rio Grande do Sul, Brazil. This program develops vigilant actions on children's health in their first year of life. It is about a qualitative descriptive exploratory research, carried out with fifteen individuals, by individual semi structured interviews. The information was analyzed according to the technique of Content Analysis. The perception of the families on the activities of the nurses was presented in two categories: establishment of a helping relationship with the users, and orientations that collaborate with the development of the children. In the opinion of the families, the activities of the nurses were performed by a qualified professional which actually satisfies the premises of health promotion. PMID- 20027954 TI - [Diabetes mellitus and the use of syringes and needles]. AB - Among the diseases that produce a great amount of garbage of health, the individuals bearers of diabetes insulino-dependent mellittus are important producers of solid garbage of health. The main objective of this study is to identify how the insulino-dependent diabetics do the discard of the syringes and needles used in their residences. The method of the research was the descriptive study. The researched population was constituted of 70 insulino-dependent individuals. The collection of data was accomplished through questionnaire, in the period of October of 2006 to March of 2007. Inside of the relevant results, in which 100% do not know what biological garbage is and 51.43% discard in the common garbage the needles and syringes and ignore the course of the garbage to the embankment, the importance is demonstrated by becoming aware and educating the population of the risks of this practice being fundamental for the environment. PMID- 20027955 TI - [Nurse and Psychosocial Attention Center's users informal caregiver: expressed health need]. AB - The informal caregivers of mental suffering persons may present healthcare needs, by living together, such as conflicts in day life, crisis situations, social discrimination, guilt, and others. This research aims to comprehend the healthcare needs of Psychosocial Attention Center's (PSAC) users' informal caregiver. It is a descriptive research. Nine informal caregivers who take part of a PSAC's family support group were interviewed, using Alfred Schutz's social phenomenology. After analyzing the interviews, two categories have emerged: "Learning" and "Experience Exchange". It was evidenced that the informal caregiver aims knowledge by exchanging experience among each other, and articulating their practical knowledge with the health professionals' theoretical knowledge. It was concluded that the healthcare needs of PSAC's users' informal caregiver must base nursing practice as conceiving health education as a reflexive action, in order to value this clientele's knowledge and experience. PMID- 20027956 TI - [Education of nurses under the social enterprising view]. AB - The purpose of this qualitative study is to discuss the education of nurses under the social enterprising view. The Grounded Theory was used as a methodological reference and interviews to collect data. The sample was composed of 35 subjects that were interviewed in different states of Brazil, which were distributed in different sample groups. Among the interviewed, there were: 20 nurses, 4 doctors, 1 dentist, 1 nutritionist, 1 pharmacist, 2 psychologists, 1 theologian, 2 teachers, 1 administrative manager and 2 young participants of the social project. Data codification led to the category: Incrementing the professional education of the nurse which will be discussed under the social enterprising view. To consider the social enterprising education of nursing, recognized as a new paradigm of social intervention, capable of potentiating the social actions through the interactive nets, it means to carry out new possibilities, for the creativity and innovation, without getting unconsidered the emerging social contradictions and without the pretension of arriving at definitive syntheses. PMID- 20027957 TI - [Research groups in nursing in the South Region part of Brazil]. AB - The study purpose is to describe how the Research Groups of Nursing Education (RGNE) are organized in the Southern of Brazil. It's a documental study descriptive and explorative, of qualitative research, with the data collection from the Brazilian National Agency for Science and Technology (CNPq) Groups Directory website 2006 census. The South Region presents 18 RGNE with 173 researchers; most of them have Master and Doctor Degree, working like professor. There are 111 students; most of them are graduating from the nursing course. Nevertheless, we still have a small number of scientific initiation scholarships. There are 27 technicians, who have got masters degree or have taken a specialization and have worked on assistance and teaching working in nursing area. The expressive participation of undergraduate students seems to improve the nursing researcher formation. Therefore, there is a great need to invest in the interdisciplinary process and the teaching-working integration at RGNE. PMID- 20027958 TI - [Impact of depressive symptoms in the quality of life of basic health care service users]. AB - This paper aims at identifying symptoms of depression in primary health care service users, assessing the association between sociodemographic characteristics and the impact of those symptoms on the quality of life. It is a transversal study with a consecutive sampling of ninety-five patients of a primary health care unit, Beck Depression Inventory (BDI) and WHOQOL-Bref were used to measure depressive symptoms and quality of life, respectively. No significant association was found among sociodemographic data, depression symptoms, and quality of life. Nevertheless, forty-four patients (46%) had depression symptoms associated to a significantly worse quality of life in all areas (p < 0.001). This finding shows the importance of being aware of the presence of depression, even in primary health care service users, considering the negative impact that depression symptoms have on the quality of life. PMID- 20027959 TI - [Nurses' opinion about nursing care quality assessment indicators]. AB - This descriptive quantitative study aimed at identifying the opinion of nurses from an accredited hospital about care quality assessment indicators. The study was carried out with nurses from a general hospital that maintains a quality program, in the countryside of the state of Sao Paulo, Brazil. Results show that indicators related to specific nursing care processes are highly valued, as well as indicators related to client satisfaction with nursing and technical staff dimensioning. Not all nurses considered indicators related to general care results as relevant for care quality assessment, or management indicators. Results show the need to involve all nurses in the development and collection of indicators, as well as to develop specific indicators for each nursing practice area. PMID- 20027960 TI - [The work in the Psychosocial Support Centers: a study in Cuiaba, Mato Grosso, Brazil]. AB - Analyzing the working conditions of Psychosocial Support Centers (CAPS) teams was the objective of this exploratory-descriptive study based on a historical critical approach that interviewed 47 professionals from five CAPS in Cuiaba, Mato Grosso, Brazil. The analysis identified: professional background and training, employment relationship, remuneration and collective organization. Most of the professionals didn't have any specialization in the area; the statutory relationship was predominant in the state public health system and the temporary ones in the city public health system, where 96% earned up to two monthly minimum wages; most of them were not unionized. We consider that those conditions cause difficulties for the attainment of the objectives of the psychosocial support. PMID- 20027961 TI - [A social poetic of the self-care: nursing students behavior aiming the promotion of health]. AB - Based on the principle that the undergraduating nursing students are future professionals of caregiving, this study aims at analyzing their understanding in relation to the care with themselves, as well as at identifying the care that they have or would like to have with their own body. To promote an effective participation of the eleven subjects, this study was based on the social and poetic method, making a research-team. Considering the self-care from the Body Dynamics as Minimum Territory, the nursing students showed analytics categories as: The self-care conscience; The wish for caregiving to spiritual, intellectual and physical body, which in the classificatory study was named "Disregard versus wish for self-care". Thus, we think that it is fundamental for nursing professionals, as well as for nursing students, to consider the self-care thematic, since the engagement with their own health is essential for the well being and best quality of life. PMID- 20027962 TI - [Experience of intensive care unit nurses dealing with death and the dying processes]. AB - This paper aims at understanding the way nurses experience their patient's death and their patients' dying processes. The researched subjects were nurses that have worked in an Intensive Care Unit of adults. Making use of the phenomenological methodology approach, the data was obtained through a semi structured interview. The analysis followed the phenomenological approach in the fields of description, reduction and understanding. From such analysis the following categories emerged: the therapeutic obstinacy as a disturbance in the death and dying processes; the hard coexistence among a group of professionals with multiple functions, death and the dying processes; the work dynamics and its implications in death and the patients' dying processes. The professionals need to comprehend that death and dying are an inalienable part of the life and not a challenge to be always overcome. PMID- 20027963 TI - Work ability and low back pain among workers from a public health institution. AB - This study investigated the association among low back pain and the work ability. A cross-sectional study was developed covering all workers from a public health institution (n = 651). They answered a questionnaire on demographics, work, lifestyle characteristics and work ability. The evaluation of work ability was developed with the work ability index (WAI). An univariate logistic regression analysis was carried out having the presence of the studied disease as a dependent variable and showed association in relation to the following WAI items: low work ability in relation to physical demands of the work, high estimated work impairment due to diseases, high sickness absence, low own prognosis of work ability two years from now and poor mental resources. The presence of low back pain affected several aspects of the work ability. PMID- 20027964 TI - [Cumulative trauma disorders: work or professional disease?]. AB - This study aimed at reviewing the Brazilian legislation applied to occupational health. It refers to the diseases embodied in the Repetition Strain Injury (RSI) and Cumulative Trauma Disorders (CTD) regarded as work or professional diseases. This analysis allowed to perform the historical evolution of legislation concerning the issue, noting that the state of the art of regulation on RSI-CTD is anchored in specific regulation present in the Normative Instruction 98/2003, that establishes the diagnostic criteria and classification of RSI-CTD. It was concluded that according to the existing legislation in Brazil, the pathologies related to RSI-CTD are considered as work diseases and their legal effects are similar to the work-related accidents. PMID- 20027965 TI - [The extra training of nursing care in the teaching hospitals]. AB - Study type integrative review of literature that aimed at understanding the integration of middle level nursing trainees in the nursing care of teaching hospitals. Using the theoretical framework of contextual analysis, in which the context is featured in four distinct interactive levels, and as containing meanings that facilitate the description and understanding of issues surrounding the phenomenon. We found thirteen texts which were organized in three levels of contextual issues: the nursing care under the immediate context; the working trainees, under the specific context; and the teaching hospital, as the metacontext. The study helped to identify the extracurricular work experience as a means of scientific and technical development of trainees in the nursing care provided in teaching hospitals, while involved in the context of the current professional world that may also be a silenced form of the precariousness of work. PMID- 20027966 TI - [Nursing care in the use of electrosurgery]. AB - Electrosurgery is a largely used technology in surgical wards. The implementation of evidence-based practice has become essential to nurses' decision making and was chosen as the adopted theoretical-methodological framework. The present integrative literature review aimed to search for and evaluate the scientific knowledge produced on nursing care related to the use of electrosurgery in the intraoperative period. Articles were selected using the databases MEDLINE and CINAHL, and 21 articles composed the sample. Results indicate that it is imperative for perioperative nurses to have the technical-scientific knowledge about the use of electrosurgery. Found evidences support the implementation of policies and procedures that assure the safety of patients, minimizing potential risks in the use of this technology. PMID- 20027967 TI - [Cardiorespiratory arrest of the adult patient in a hospital environment: nursing contributions]. AB - The objective of this study was to analyse the scientific production regarding cardiorespiratory arrest (CA) in adult patients in a hospital environment, as to support nursing knowledge. This is a systematic review, which were used SciELO, LILACS and MEDLINE databases from 1997 to 2007. Most of the articles used in the analysis referred to the survival prognostic factors of patients submitted to cardiopulmonary ressucitation (CPR) and to the results of this intervention. The prospective studies were methodologically prevalent. The nursing scientific production was numerically small when compared to the medical area. The importance of a nursing team trained and apt for CA treatment and prevention was pointed out; continual education programs were stimulated. The conclusion is that there are few nursing studies about CPR, and continual incentive to the scientific production regarding its acting both in CA prevention and RCP management is imperative. PMID- 20027968 TI - [The gift theory and care giving in nursing]. AB - This is a reflection on the interpersonal relations established between patients and the nursing team with emphasis on the configuration of the bonds resulting from such relationship, theoretically supported by Marcel Mauss's gift theory. Special consideration is given to the person-to-person interaction as the main component of the structure on which the actions performed by the health professionals find support, paving the way to interpersonal development aiming at making it easier to have a good performance of the services rendered and even going beyond technical matters. Based on the gift theory, it is possible to infer that symbolic assets are in circulation between the nursing team and the patients, in health care, demanding from the professionals humanitarian postures that are to be construed in terms of gestures, behaviors and attitudes. PMID- 20027969 TI - [Trajectory of a research on the HIV/AIDS theme: limits and (im) possibilities]. AB - The article presents the trajectory of a research group upon the development of the project funded by the Health Ministry, through the National HIV/AIDS/DST Program, named "Impact of adhesion to the anti-retroviral among children and adolescents under the perspective of the family, the child and the adolescent in the municipalities of Porto Alegre andSanta Maria, RS". With the objective of presenting the stages of the research, from its planning up to the phase of data collection, the study was carried out in two municipalities simultaneously, counting on 20 participants comprising researchers and scholarship students. The stages developed until the data collection were: inclusion of care and teaching professionals in the research nucleus; selection and training of scholarship students; elaboration of the instrument; elaboration of a manual team habilitation; application of the pilot test. PMID- 20027970 TI - [The use of the Delphi technique for competencies investigation: an experience report]. AB - The Delphi technique has been used in nursing research, with electronic means as an option. This article has the objective of reporting researchers' experience on the application of this technique in a study about nurses' competencies for the performance of Essential Public Health Functions. Experts were searched on the Lattes Platform of the National Council of Scientific and Technologic Development (CNPq) in Brazil. Two groups with 50 participants each, one with nurses and other with faculty with representation of every geographic region of the country were organized. The competencies validation instrument was sent by electronic means. Although it is reported that this form reduces the time for sending and receiving the questionnaires, difficulties to obtain contact with the experts, lack of return of the participants and a long time to answer the instrument were observed. Health care professionals need to understand the importance of their participation in researches with the utilization of this technique. PMID- 20027971 TI - [Influenza A/H1N1 in Israel--clinical and epidemiological characteristics]. AB - BACKGROUND: In March 2009, a novel A/H1N1 influenza virus began its inexorable spread around the world. Information regarding disease characteristics, groups at risk and prognosis remain partial. The Epidemiology Division of the Israeli Ministry of Health performs ongoing influenza surveillance and tracking of patients in Israel. The authors set out to characterize the disease and its spread in Israel. METHODS: Surveillance and investigation procedures were modified in accordance with changing Ministry of Health policy. From the outset of the outbreak and until June 30, 2009, all suspected cases of influenza A/H1N1 were investigated and laboratory verified. Starting July 1, 2009, lab confirmation was reserved for severely ill patients or those at high risk of complications. All hospitalized cases were monitored and tracked daily. RESULTS: By June 30, 2009, 596 patients had laboratory confirmed Influenza A/H1N1: 58% of these were aged 10-30 years, and only 5% were above 50 years of age; 58% were male. In addition to fever (83%), patients reported cough (74%), rhinorrhea (59%), and headache and sore throat (53% each). Thirty three patients were considered at high risk for complications, four of which required hospitalization in an intensive care unit; 64% of infections were acquired in Israel and 22% in the United States. By July 29, 2009, 952 additional cases had been verified. Overall, 13 of the cases had been hospitalized in intensive care, 3 of whom died. DISCUSSION: Early data indicate spread particularly to younger populations, expressing non-specific respiratory symptoms. Ongoing investment in real-time data collection and analysis will enable epidemiologists to supply the information necessary to deal with the influenza epidemic. PMID- 20027972 TI - [743 New York Emergency Room rotation under the threat of swine flu]. PMID- 20027973 TI - [Adult pertussis poses a severe risk for infants]. AB - In Israel, there have been increasing reports of Bordetella pertussis infection among adolescents and adults, but the peak incidence and highest mortality occur among infants. The authors report four cases involving the likely transmission of pertussis from parents to their offspring in two hospitals in Israel. The adoption of proper infection control measures and targeted screening of parents may reduce the potential risk for such transmission. PMID- 20027974 TI - [The burden and outcomes of acute bronchiolitis among young children hospitalized in Israel]. AB - BACKGROUND: Acute bronchiolitis (AB) is a significant indication for hospitalization during the winter period. Underlying conditions increase risk for severe manifestations. AIMS: To estimate the burden and outcomes of AB in northern Israel. METHODS: A prospective study was performed between 1.12.2005 - 31.3.2006. Previously healthy children younger than 2 years of age, hospitalized with the diagnosis of AB, in three hospitals in northern Israel, were included in this study. RESULTS: Overall, 465 children (93%) out of 500 children who were hospitalized due to AB and comprised 18% of all hospitalizations during the study period, were included. A pathogen was identified in 91% of cases. A single pathogen was identified in 243 (52%) cases; 2-4 pathogens were found in 176 (39%) children. Common pathogens were respiratory syncytial virus (RSV) and Rhinovirus in 346 (75%) and 129 (28%), of which 192 (41%) and 37 (8%) were sole pathogens respectively. Complete data were available for 390 (82%) children, of whom 78% were younger than 6 months. Patients were hospitalized for 4 +/- 4.4 days; 15 children were treated in intensive care. There was one mortality. An X-ray was performed in 94% of cases. Most children were treated with multiple regimens including intravenous fluids, O2 supplement, physiotherapy, bronchodilators, steroids and antibiotics. Bronchiolitis was attributed to 18% of hospitalizations. On a national scale, assuming that the policy of hospitalization is similar, 4100 children were hospitalized due to AB. This reflects 4% of all hospitalized children a year in Israel and attributing to 16,400 hospitalizations. CONCLUSIONS: AB is associated with a huge burden, during a short period of the year, on pediatric departments, mainly attributed by infants younger than 6 months of age. The development of effective vaccine against RSV may significantly reduce the burden of morbidity. PMID- 20027975 TI - [The relationship between fever magnitude and serious bacterial infections in febrile infants less than two-months-old--a prospective study]. AB - BACKGROUND: The evaluation and treatment of febrile infants below 2 months of age is inconsistent in hospitals in Israel. In some centers fever magnitude is considered to be one of the parameters that influence decision making as well as management. OBJECTIVE: To assess the correlation between fever magnitude and the risk of serious bacterial infections (SBI) in hospitalized infants under the age of 2 months. PATIENTS AND METHODS: The study group consisted of all infants < or = 2 months hospitalized at the Schneider Children's Medical Center for evaluation of fever from September 2006 to December 2008. Data were collected prospectively regarding the magnitude of fever on admission, during hospitalization, and diagnosis of SBI. RESULTS: SBI was detected in 90 (10.8%) of the 833 infants that met the inclusion criteria (UTI in 68, bacteremia in 11, pneumonia in 10 and enteritis in one). The mean fever at presentation was 38.4 degrees in the group of infants with SBI compared to 38.3 degrees in the infants without SBI (p=NS). Mean maximal fever during hospitalization was 38.6 degrees in the two groups. No correlation was found between the degree of fever and the type of SBI. CONCLUSION: There is no correlation between fever magnitude and the risk of SBI in hospitalized infants aged < or = 2 months. Therefore, we suggest that fever magnitude should not be a factor in the decision of evaluating febrile infants in this age group. PMID- 20027976 TI - [Mycobacterium abscessus infection in insulin injection sites in a diabetic patient]. AB - Insulin injection site abscesses have been attributed to pyogenic streptococci and Staphylococcus aureus. In this report, the authors describe a patient in whom Mycobacterium abscessus was isolated from one of multiple insulin injection sites. The article reviews six additional cases of rapidly growing Mycobacteria identified as the causative agent of skin infections at insulin injection sites. In most cases, precise identification of the source of infection was not successful. Possibly, inadequate skin preparation prior to injection or a contaminated insulin ampoule resulted in these infections. PMID- 20027977 TI - [Ill-appearing febrile 5-week-old infant: the rule of empiric treatment]. AB - This is a case study of a 5-week-old child, after prolonged hospitalization in the Neonatal Intensive Care Unit, with fever and urinary tract infection caused by extended-spectrum beta-lactamase (ESBL)-producing, gentamicin-resistant, Escherichia coli. This case highlights the importance of taking risk factors for resistant bacterial infection into account in young, ill-appearing infants, and suggests that in some cases empirical antibiotic treatment should be even broader than recommended by current guidelines. PMID- 20027978 TI - [Community-associated methicillin-resistant Staphylococcus aureus infections among children in the western Galilee region: the beginning of an epidemic?]. AB - BACKGROUND: Community-associated (CA) methicillin-resistant Staphylococcus aureus (MRSA) infections were reported in the United States in the 1990's among healthy children, without traditional risk factors for infection with MRSA. CA-MRSA is now epidemic in the United States causing superficial infections and severe invasive disease. There is a paucity of data about CA-MRSA disease in the pediatric Israeli population and the common belief is that CA-MRSA disease is rare. OBJECTIVE: Prospective surveillance for CA-MRSA strains among hospitalized children at the Western Galilee Hospital in Israel over a 1-year period. METHODS: Staphylococcus aureus isolates were collected prospectively in 2007-8. CA-MRSA strains were defined as MRSA isolated in the first 72 hours of hospitalization with non-multi-drug resistant profile (resistant to < 3 non-beta-lactam antimicrobials). Patients' clinical characteristics were collected from medical charts. RESULTS: Five of 103 (4.8%) children were identified with Staphylococcus aureus isolates that were infected with CA-MRSA strains. All five had MRSA isolated from skin abscesses; their median age was 2.5 years. Skin and soft tissue infections were evident among 59 children infected with Staphylococcus aureus, 5 of which were CA-MRSA strains (8.5%). The calculated prevalence of CA MRSA infections among pediatric patients within the study period was 76 cases per 100,000 admissions. All isolates were susceptible to vancomycin, linezolid, rifampin, trimethoprim sulfamethoxazole, and clindamycin. Four of the 5 children presented with recurrent MRSA infections. Possible risk factors for CA-MRSA were noted among 3 patients: participation in contact sports, atopic dermatitis, and family history of MRSA infections. CONCLUSIONS: A significant rate of CA-MRSA disease exists among children in the Western Galilee Region. CA-MRSA disease presents as recurrent skin and soft tissue infections. Awareness and recognition by culturing, mainly of skin lesions, is the first crucial step in investigating the magnitude of CA-MRSA disease. The old school thinking of MRSA being solely a hospital-based pathogen, in patients with frequent exposure to the healthcare system, is no longer valid in Israel. PMID- 20027979 TI - [Gastrointestinal carriage of Pseudomonas aeruginosa in children residing in chronic care institutes in Jerusalem: high prevalence and high rates of antibiotic resistance]. AB - BACKGROUND: Pseudomonas aeruginosa carriage in the gastrointestinal tract is uncommon in healthy children. Children living in chronic care institutions are often carriers of P. aeruginosa in the respiratory tract, but data is lacking regarding gastrointestinal carriage in these children. AIMS: To examine the carriage rate of P. aeruginosa in children living in chronic care institutions in Jerusalem and to assess resistance rates of the bacteria to different classes of antibiotics. METHODS: Rectal swabs were taken from all children residing in two chronic care institutions in Jerusalem: "St. Vincent" and "Aleh". The swabs were examined for presence of Pseudomonas aeruginosa. The authors used disk diffusion technique and E Test to assess resistance for different antibiotics. RESULTS: Gastrointestinal carriage of P. aeruginosa was detected in 37 out of 125 of the children (30%); 16% of the P. aeruginosa isolates were resistant to carbapenems; 16% were resistant to aminoglycosides, 14% to ureidopenicillins and 11% to quinolones. All isolates were sensitive to ceftazidime and colistin. In 84% of the isolates, the minimal inhibitory concentration (MIC) for meropenem was significantly lower than the MIC for imipenem. SUMMARY: P. aeruginosa is a common colonizer of the gastrointestinal tract of children living in chronic care institutions. Empiric antibiotic treatment against P. aeruginosa should be considered when treating children with acute gastrointestinal pathologies. Antibiotic resistance, and particularly carbapenem resistance, is common in this population. There is a significant difference between the MICs for imipenem and meropenem. Future studies are needed to understand the clinical significance of this finding. PMID- 20027980 TI - [Hinman syndrome--an uncommon cause of recurrent urinary tract infections since childhood]. AB - A 20-year-old female presented with a history of recurrent urinary tract infections, diurnal incontinence, nocturnal enuresis, and voiding difficulty. Imaging studies revealed a thick trabeculated bladder and high post-void residual urine volumes. Urodynamic evaluation demonstrated involuntary bladder contractions and no evidence of bladder outlet obstruction. A full neurological and urological workup were both unremarkable. A thorough history revealed her symptoms were initiated after prolonged and stressful potty training with multiple failed attempts. She developed a dysfunctional voiding pattern consistent with a non-neurogenic neurogenic bladder, also known as the "Hinman syndrome". Successful therapy relies primarily on behavior modification maneuvers combined with medical therapy to reduce unstable bladder contractions and alleviate bladder outlet obstruction. PMID- 20027981 TI - [Vibrio vulnificus infection outside of the fish ponds in northern Israel: acquisition in the "Einot Tzukim" springs near the Dead Sea]. AB - Vibrio vulnificus is a Gram-negative bacterium that may cause severe skin and systemic infection after exposure of open wounds to contaminated water, especially in patients with underlying disease such as immune-deficiency, iron overload or end stage liver or renal disease. The V. vulnificus infection has been reported in Israel almost exclusively after exposure to Tilapia fish cultivated in fresh water fish ponds in northern Israel. The authors report the first case of V. vulnificus infection acquired in a nature reserve in southeastern Israel, with no connection to fish handling. A 14.5-years-old girl with transfusion-dependant thalassemia major presented with high fever and a rapidly progressive bullous cellulitis of the ankle. The infection occurred around a cut on the left lateral malleolus, after bathing in the fresh water ponds of Einot Tzukim (Ein Feshcha) in south-eastern Israel, and progressed despite the use of broad-spectrum antibiotics. Blood and wound cultures eventually yielded Vibrio vulnificus and appropriate treatment was commenced. The fever subsided after a few days but resolution of the local findings was very gradual and lasted for weeks. The presence of V. vulnificus in natural springs far from the northern artificial fish ponds broadens the danger of this infection. We find it prudent to advise people at risk for V. vulnificus infection, such as those suffering from immunedeficiency, iron overload and end stage liver or renal disease, to refrain from bathing in natural ponds whilst injured. PMID- 20027982 TI - [Infectious diseases--the progress is huge--a lot remains to be done]. AB - Huge progress has been achieved in the last century in preventing and treating infections. The discovery of antibiotics and the ongoing development of appropriate vaccines have decreased the incidence of many infectious diseases. This led Sir McFarland Burnett to state in 1962 that 'By the end of the Second World War it was possible to say that almost all of the major practical problems of dealing with infectious disease had been solved.' More recently, however, new challenges have emerged: antibiotic resistance is spreading in hospitals as well as in communities, nosocomial infections are both a health problem and economic burden, and new infections have been emerging while others are reemerging. The articles included in this issue of Harefuah, which is dedicated to infectious diseases, exemplify the continuing challenges presented to the medical community by these diseases. PMID- 20027983 TI - [The diagnosis and therapy of first community acquired urinary tract infection in children]. AB - A first urinary tract infection (UTI) in childhood is more prevalent in females < 5-years-old. Circumcision generally protects males from UTI, however, during the month following the procedure, the prevalence of infection increases up to 12 times in circumcised boys when compared with those not circumcised. Almost all the infections are caused by aerobic Gram-negative bacteria of which E. coli are responsible for 70-90% of the cases. Signs and symptoms of UTI vary in different age groups. Factors associated with the likelihood of UTI are: non-circumcised male, fever > 40 degrees C, and a fever > 39 degrees C for more than 48 hours with no other focus of infection on physical examination. Urinalysis and urine microscopy are screening tests for UTI. In children with clinical symptoms and signs suggesting UTI, the results of these tests have a positive predictive value (if both are positive), or negative predictive value (if both are negative) approximating 100%. The definitive diagnosis of UTI is based on the urine culture. Bag urine culture is associated with a very high rate of contamination. Therefore, in non-toilet trained children, urine culture should be obtained directly from the urinary bladder either by supra pubic aspiration or in and out transurethral catheterization. Mid stream clean voided urine specimens obtained from circumcised males in the first months of life are also acceptable. Depending on the clinical presentation, oral therapy can begin from as early as two months of age, and the recommended empiric drugs for first febrile UTI are cefuroxime axetil, or amoxicillin clavulanate. Cephlexin is recommended for cystitis. PMID- 20027984 TI - [Clostridium difficile associated diarrhea (CDAD)--hard to crack]. AB - Clostridium difficile-associated diarrhea (CDAD) is the most common documented cause of nosocomial diarrhea in developed countries. In recent years morbidity and mortality associated with this pathogen have increased significantly, simultaneously with the appearance of a new strain--NAP1/BI/027--which was implicated in epidemics of severe disease in hospitals around the world. Despite the accumulated experience in identification and treatment of patients, physicians continue to face complicated problems concerning the diagnosis, treatment and prevention of CDAD. However, significant effort is invested in the development of more accurate diagnostic tests, newer drugs for treatment and new methods to prevent the spread of the pathogen. The problems related to CDAD and updated possible solutions are discussed in this article. PMID- 20027985 TI - Pupil size, white-to-white corneal diameter, and anterior chamber depth in patients with myopia. AB - PURPOSE: To evaluate anatomical parameters in a population of patients with myopia. METHODS: Nine hundred sixty-four myopic eyes (-3.00 to -20.00 diopters [D] spherical equivalent refraction) were evaluated to measure mesopic and photopic pupil size with an infrared pupillometer; anterior chamber depth and white-to-white corneal diameter were obtained with Orbscan II (Bausch & Lomb). Correlation analysis was performed to evaluate the relationships among anatomical parameters of the anterior segment of the eye. RESULTS: Average change in pupil size between mesopic and photopic conditions shows a uniform gap of 1.5 mm in patients aged 18 to 62 years with a slight insignificant trend to decrease with age. Photopic and mesopic pupil size were highly correlated (r=0.694, P<.001) and the difference between both measures was positively correlated with mesopic pupil size (r=0.207, P<.001) and inversely correlated with photopic pupil size (r=0.561, P<.001). Anterior chamber depth and white-to-white corneal diameter were positively correlated (r=0.389, P<.001). White-to-white corneal diameter and anterior chamber depth were not correlated with age (r=-0.096, P<.001) or anterior chamber depth (r=-0.183, P<.001-0.183) as a function of age. CONCLUSIONS: Average difference between photopic and mesopic pupil size remained constant across the range of ages included in this cohort. A positive correlation was noted between anterior segment dimensions, and anterior chamber depth decreased with age. PMID- 20027986 TI - US FDA clinical trial of the Tetraflex potentially accommodating IOL: comparison to concurrent age-matched monofocal controls. AB - PURPOSE: To assess the efficacy of the Tetraflex (Lenstec Inc) intraocular lens (IOL) to provide enhanced near reading ability and spectacle independence relative to a monofocal control IOL in bilaterally implanted eyes tested binocularly. METHODS: A prospective, age-matched, non-randomized US Food and Drug Administration clinical trial of 255 Tetraflex and 101 monofocal IOL control patients was performed. To date, 239 Tetraflex and 96 control patients were examined at 12 months postoperatively. RESULTS: At 12 months postoperative, the Tetraflex patients read better than the controls at print sizes of 20/80 (P=.04), 20/63 (P=.01), 20/50 (P<.001), 20/40 (P=.001), 20/32 (P<.001), and 20/25 (P=.001). The proportion of patients reading at a speed of >=80 words per minute was significantly higher with the Tetraflex IOL (P=.003). Ninety-six percent of Tetraflex patients reported never wearing glasses for distance compared with 80% of control patients (P<.001). Seventy-five percent of the Tetraflex patients reported near spectacle wear either never or only occasionally for small print and/or dim light (21% never) compared with 46% of control patients (P<.001) (9% never). Near add power requirement for corrected near visual acuity was less in the Tetraflex group (P<.001); 28% of Tetraflex patients required <=1.25 diopters of near add, compared to only 7% of control patients. Spectacle independence, as measured by the proportion of patients with uncorrected distance visual acuity of 20/25 or better and various degrees of uncorrected near visual acuity, was also significantly better (P<.001) as was distance-corrected near visual acuity (P<.001). CONCLUSIONS: The results support the efficacy of the Tetraflex IOL to provide enhanced near reading ability and spectacle independence relative to a monofocal IOL control. PMID- 20027987 TI - Determining spherocylindrical correction using four different wavefront error analysis methods: comparison to manifest refraction. AB - PURPOSE: To evaluate methods that determine refractive correction from wavefront data that best matches the manifest refraction. METHODS: Wavefront data in the form of Zernike polynomial coefficients from several VISX US Investigational Device Exemption studies were used to calculate the spherocylindrical distance correction using four methods: full pupil second order, central curvature, reduced aperture, and 4-mm pupil. The data were analyzed in two groups, preoperative (3686 measurements, 882 eyes) and 6-month postoperative (2570 measurements, 797 eyes). Differences were formed between values found with each method and the manifest refractive values for the spherical equivalent (dSE), cylinder power (dCc), and axis power (dAc). The wavefront spherical equivalent values were corrected for chromatic shift, lane length, and vertex. RESULTS: Values derived using the reduced aperture and 4-mm pupil methods corresponded with manifest spherical equivalent values for both preoperative (x(-) = -0.03 diopters [D], x(-) =0.95 D) and postoperative (x(-) = -0.36 D, x =0.65 D) data. The full pupil second order method results corresponded best with manifest data for the astigmatic variables for both preoperative (x(-) dCc=0.15 D, sigma dCc=0.31 D, x(-) dAc = 0.02 D, sigma dAc =0.22 D) and postoperative data (x dCc=0.25 D, sigma dCc=0.33 D, x dAc=0.01 D, sigma dAc=0.21 D). However, all methods did well on these variables. CONCLUSIONS: The Zernike coefficient set as measured best calculates the astigmatic correction. The Zernike coefficient set resized to a 4-mm pupil is best in determining spherical equivalent. PMID- 20027988 TI - Quantitative assessment of bandage soft contact lens wear immediately after LASIK. AB - PURPOSE: To evaluate the efficacy and tolerability of overnight bandage soft contact lenses applied immediately after LASIK. METHODS: Bandage soft contact lenses were applied immediately following bilateral simultaneous LASIK in 161 consecutive patients. Signs and symptoms associated with contact lens wear were prospectively recorded at 1 hour, 4 hours, and the following morning, using a standardized questionnaire. Contact lenses were removed at 1 hour if patients were symptomatic of intolerance. If no signs of contact lens intolerance appeared at 1 hour, the lenses were removed the following morning. No contact lenses were applied in 61 patients after LASIK. RESULTS: One hour after the procedure, contact lenses were removed from 47 patients due to intolerance. In these patients, contact lens removal was associated with a significant decrease of the total score the next morning (0.31+/-0.55) compared to 1 hour (5.65+/-1.60, P<.0001). A similar decrease of the total score was noted in those patients who tolerated the contact lenses (0.20+/-0.52 vs 1.30+/-1.78) and those in whom the contact lenses were not applied (0.76+/-1.19 vs 3.24+/-1.93). Mucoid discharge at the lid margins was noted in 52.6% of patients and mild corneal edema was noted in 28.8% of patients with bandage soft contact lenses at 12 to 18 hours. CONCLUSIONS: Bandage soft contact lens application after LASIK is associated with significant reduction of symptoms during the immediate postoperative period. However, removal of these lenses may be necessary shortly after the procedure. PMID- 20027989 TI - LASIK flap buttonhole treated immediately by PRK with mitomycin C. AB - PURPOSE: To describe the visual outcomes of three patients who had LASIK flap buttonhole and were treated immediately with photorefractive keratectomy (PRK) and topical mitomycin C (MMC) 0.02%. METHODS: Three patients underwent bilateral LASIK with the SCHWIND Carriazo-Pendula 90 microm head microkeratome. In all three cases, a buttonhole flap occurred in the left eye. The flap was repositioned and phototherapeutic keratectomy for 50 microm was used for epithelial removal while immediate PRK with MMC was performed to treat the buttonhole flap. RESULTS: Three months after the procedure, uncorrected distance visual acuity and corrected distance visual acuity were 20/20 with regular topographic findings. CONCLUSIONS: Using PRK with MCC immediately after the occurrence of the LASIK flap buttonhole may be an effective treatment. PMID- 20027990 TI - Traumatic flap striae 6 years after LASIK: case report and literature review. AB - PURPOSE: To report a case of traumatic flap striae without flap dislocation 6 years after LASIK and provide a literature review of surgical flap striae, late traumatic flap striae, and their management. METHODS: A 28-year-old man presented with late traumatic flap striae without concurrent flap dislocation, which closely approximated the longest reported interval between LASIK and the development of flap striae. RESULTS: In the absence of flap dislocation, the finding of striae alone was subtle and went undetected initially. The flap was successfully refloated, stretched, and smoothed with recovery of 20/20 vision. CONCLUSIONS: Traumatic LASIK flap complications may occur many years after the original procedure. This report presents the first case of late traumatic flap striae without concurrent flap dislocation. Proper management can restore good visual function. PMID- 20027991 TI - Effects of decentration of photorefractive keratectomy on the induction of higher order wavefront aberrations. AB - PURPOSE: To evaluate the effects of ablation decentration on the induction of higher order wavefront aberrations (HOAs) in active eye-tracker-assisted myopic photorefractive keratectomy (PRK) using the VISX STAR S4 laser with ActiveTrak (Abbott Medical Optics [AMO]). METHODS: Ninety-four myopic eyes (53 patients) were divided into three groups according to ablation decentration (group 1, <=0.15 mm, 20 eyes; group 2, >0.15 to <=0.30 mm, 54 eyes; and group 3, >0.30 mm, 20 eyes). The distances of ablation centers from the centers of the entrance pupils were analyzed using corneal topography. Wavefront errors were measured preoperatively and at 6 months after PRK using a VISX WaveScan aberrometer (AMO). Statistical analysis was performed to assess the influence of ablation decentration on PRK-induced HOAs. RESULTS: The mean decentration was 0.23+/-0.10 mm (range: 0.04 to 0.52 7 mm). The magnitude of all HOAs was significantly increased at 6 months postoperatively (P<.05). Increases in PRK-induced HOAs including total HOA, coma, and spherical aberration were significantly different among the three groups (P<.05). Statistically significant differences were noted between pairs of data in group 1 versus group 3 for total HOA (P=.015), coma (P=.038), and spherical aberration (P=.038), and group 2 versus group 3 for coma (P=.049). CONCLUSIONS: Ablation decentration >0.30 mm from the center of the entrance pupil was associated with greater induction of total HOA, coma, and spherical aberration after PRK, as compared with ablation decentration <0.15 mm. In addition, ablation decentration has a more significant influence on coma inducing effects. PMID- 20027992 TI - Occupational health nursing practice through the Human Caring lens. AB - Many health care and academic centers have adopted Watson's Theory of Human Caring as their guiding principle; the theory is also used in other disciplines, such as library science. Human caring theory offers occupational health nurses as structure that not only defines a focus for practice, but also provides a basis for moral and philosophical practice analyses. In particular, nurses may find this theory useful in confirming the definition of "caring" and reconsidering what nursing is all about. More importantly, consideration and application of this theory may lead to research on its applicability to the field of occupational health nursing. This article presents the science and philosophy of human caring, specifically Watson's Theory of Human Caring. Two case studies are presented that demonstrate how the theory could be used to evaluate occupational health nursing practice. To demonstrate its possible relevance as an occupational health nursing framework, an analysis of and comparison to existing occupational health nursing guidelines are detailed and discussed. PMID- 20027993 TI - Hispanic employees in the workplace: higher rate of fatalities. AB - This article reviews the higher fatality and injury rates among the Hispanic population in the United States, whether legal immigrants, citizens, or illegal immigrants; reviews the current government and private industry regulations and safety programs; proposes additional legislation or programs; and describes the role of the occupational and environmental health nurse in reducing injuries and fatalities in this population. PMID- 20027994 TI - Microscopic composition maps of poly(styrene-co-2-hydroxyethyl methacrylate) colloidal crystals and interconnected colloidal arrays. AB - Colloidal crystalline films were prepared from poly(styrene-co-2-hydroxyethyl methacrylate) (PS-HEMA) latex particles by evaporative deposition. The hexagonally ordered surfaces of the colloidal crystals (CCs) were transformed with styrene vapor at room temperature to interconnected colloidal arrays (ICAs) that have a honeycomb-like ridge of polymer surrounding hexagonally ordered dimples in the surface. When the styrene vapor temperatures were increased systematically to 45 degrees C, the regularity of ICA structure decreased and finally disappeared. Images from transmission electron microscopy (TEM) and from atomic force microscopy (AFM) show that the surfaces of the PS-HEMA particles and the ICAs have raspberry textures. Monolayer CCs and ICAs fabricated on TEM grids were analyzed by energy dispersive spectroscopy (EDS) to determine the elemental compositions of the different regions of the textured surfaces. Carbon, oxygen, and sulfur were distributed all over the surface of the CC. While carbon was distributed over the entire surface of the ICA, oxygen, sulfur, sodium, and potassium were concentrated mainly on the ridges of the honeycomb and not in the dimples of the ICA. The results are discussed in terms of a mechanism of transformation of the CC to the ICA in which styrene monomer swells the polystyrene-rich regions of the particles, and the swollen polystyrene rises to the surface. The polyHEMA-rich regions of the particles maintain the hexagonal periodicity, and liquid styrene evaporates to leave a more polystyrene-rich textured surface. PMID- 20027995 TI - Salts-based size-selective precipitation: toward mass precipitation of aqueous nanoparticles. AB - Purification is a necessary step before the application of nanocrystals (NCs), since the excess matter in nanoparticles solution usually causes a disadvantage to their subsequent coupling or assembling with other materials. In this work, a novel salts-based precipitation technique is originally developed for the precipitation and size-selective precipitation of aqueous NCs. Simply by addition of salts, NCs can be precipitated from the solution. After decantation of the supernatant solution, the precipitates can be dispersed in water again. By means of adjusting the addition amount of salt, size-selective precipitation of aqueous NCs can be achieved. Namely, the NCs with large size are precipitated preferentially, leaving small NCs in solution. Compared with the traditional nonsolvents-based precipitation technique, the current one is simpler and more rapid due to the avoidance of condensation and heating manipulations used in the traditional precipitation process. Moreover, the salts-based precipitation technique was generally available for the precipitation of aqueous nanoparticles, no matter if there were semiconductor NCs or metal nanoparticles. Simultaneously, the cost of the current method is also much lower than that of the traditional nonsolvents-based precipitation technique, making it applicable for mass purification of aqueous NCs. PMID- 20027996 TI - Effect of confinement on the mesoscale and macroscopic swelling of thin block copolymer films. AB - We report on the swelling behavior and the corresponding morphological behavior of cylinder-forming polystyrene-b-polybutadiene diblock copolymers, which are confined to several layers of structures. The equilibration of thin films has been done under controlled atmosphere of a nonselective solvent. In situ spectroscopic ellipsometry measurements revealed more than 10% increase of the solvent uptake with decreasing film thickness. With scanning force microscopy of the microphase separated patterns in quenched films, the correlation between the degree of the long-range order of cylinder domains and the degree of the macroscopic swelling has been established. In the case of spontaneously formed micrometer-sized topographic features with discreet film thickness (terraces), the increased solvent uptake by thinner films holds true even for isolated terraces on the mesoscale. The observation of nonhomogeneous swelling of the films on the micrometer scale brings novel insights into the properties of confined soft matter, and suggests new approaches toward the fabrication of polymer-based nanostructured responsive materials. PMID- 20027997 TI - Equilibrium and molecular mechanism of anionic dyes adsorption onto copper(II) complex of dithiocarbamate-modified starch. AB - The assembly of dyes molecules on metal-polymer complexes is of interest due to their potential applications in photovoltaic cell, separation, and wastewater treatment. In the present work, the interaction of anionic dyes (acid orange 7, acid orange 10, acid green 25, and acid red 18) with the copper(II) complex of dithiocarbamate-modified starch (DTCSCu) was investigated. The sorption studies showed that the interaction mechanism was based on chelating adsorption. The equilibrium data fitted well with Langmuir-Freundlich isotherm, and the capacities followed the order AO7 > AG25 > AR18 > AO10. It was affected by the structure of the dye. The sulfonate groups located on benzene rings favored efficient adsorption. Despite the difference in capacity, the molar n(dye):n(Cu) ratios for acid orange 10, acid red 18, and acid green 25 were approximately 1:2 when the maximum capacities for the dyes were achieved at the optimal pH of 4. It suggested that one dye molecule bound to one dinuclear copper center on DTCSCu. The molar n(dye):n(Cu) ratio for the smallest dye, acid orange 7 (AO7), approached 1:1, demonstrating two AO7 molecules binding to two copper ions of the dinuclear core. The dyes adsorption related to the dinuclear copper core available on the polymer was further verified by electron spin resonance studies. Such interaction resulted in the formation of a ternary dye-metal-polymer complex. The ternary complexes were more stable than DTCSCu, which favored the adsorptions. PMID- 20027998 TI - Origins of regioselectivity of Diels-Alder reactions for the synthesis of bisanthraquinone antibiotic BE-43472B. AB - The regioselectivities of several Diels-Alder reactions utilized en route to bisanthraquinone antibiotic BE-43472B are examined using density functional theory calculations. These reactions involve highly substituted dienes and juglone dienophiles, and there is an opposite regiochemical outcome for Diels Alder reactions with beta-aryl substituted juglones when compared to reactions of unsubstituted juglone. In this article, the effect of an aromatic conjugating group bonded to juglone is explored. PMID- 20027999 TI - Indium-mediated asymmetric barbier-type allylations: additions to aldehydes and ketones and mechanistic investigation of the organoindium reagents. AB - We report a simple, efficient, and general method for the indium-mediated enantioselective allylation of aromatic and aliphatic aldehydes and ketones under Barbier-type conditions in a one-pot synthesis affording the corresponding chiral alcohol products in very good yield (up to 99%) and enantiomeric excess (up to 93%). Our method is able to tolerate various functional groups, such as esters, nitriles, and phenols. Additionally, more substituted allyl bromides, such as crotyl and cinnamyl bromide, can be used providing moderate enantioselectivity (72% and 56%, respectively) and excellent diastereoselectivity when employing cinnamyl bromide (>95/5 anti/syn). However, the distereoselectivity when using crotyl bromide was poor and other functionalized allyl bromides under our method afforded low enantioselectivities for the alcohol products. In these types of indium-mediated additions, solvent plays a major role in determining the nature of the organoindium intermediate and we observed the susceptibility of some allylindium intermediates to hydrolysis in protic solvents. Under our reaction conditions using a polar aprotic solvent, we suggest that an allylindium(III) species is the active allylating intermediate. In addition, we have observed the presence of a shiny, indium(0) nugget throughout the reaction, irrespective of the stoichiometry, indicating disproportionation of indium halide byproduct formed during the reaction. PMID- 20028000 TI - Total synthesis of uniflorine A, casuarine, australine, 3-epi-australine, and 3,7 di-epi-australine from a common precursor. AB - A flexible method for the diastereoselective total synthesis of the pyrrolizidine alkaloids uniflorine A, casuarine, australine, and 3-epi-australine and the unnatural epimer 3,7-di-epi-australine from a common chiral 2,5-dihydropyrrole precursor is described. PMID- 20028001 TI - Time-resolved gas-phase kinetic, quantum chemical and RRKM studies of reactions of silylene with cyclic ethers. AB - Time-resolved kinetic studies of silylene, SiH(2), generated by laser flash photolysis of phenylsilane, have been carried out to obtain rate constants for its bimolecular reactions with oxirane, oxetane, and tetrahydrofuran (THF). The reactions were studied in the gas phase over the pressure range 1-100 Torr in SF(6) bath gas, at four or five temperatures in the range 294-605 K. All three reactions showed pressure dependences characteristic of third-body-assisted association reactions with, surprisingly, SiH(2) + oxirane showing the least and SiH(2) + THF showing the most pressure dependence. The second-order rate constants obtained by extrapolation to the high-pressure limits at each temperature fitted the Arrhenius equations where the error limits are single standard deviations: log(k(infinity)oxirane/cm3 molecule(-1)s(-1))=(-11.03+/ 0.07) + (5.70+/-0.51)kJ mol(-1) ln 10, log(k(infinity)oxetane/cm3 molecule(-1)s( 1))=(-11.17+/-0.11)+(9.04+/-0.78) kJ mol(-1)/RT ln 10, log(k(infinity)THF/cm3 molecule(-1)s(-1))=(-10.59+/-0.10)+(5.76+/-0.65) kJ mol(-1)/RT ln 10. Binding energy values of 77, 97, and 92 kJ mol-1 have been obtained for the donor acceptor complexes of SiH2 with oxirane, oxetane, and THF, respectively, by means of quantum chemical (ab initio) calculations carried out at the G3 level. The use of these values to model the pressure dependences of these reactions, via RRKM theory, provided a good fit only in the case of SiH2 + THF. The lack of fit in the other two cases is attributed to further reaction pathways for the association complexes of SiH2 with oxirane and oxetane. The finding of ethene as a product of the SiH2 + oxirane reaction supports a pathway leading to H2Sid=O + C2H4 predicted by the theoretical calculations of Apeloig and Sklenak. PMID- 20028002 TI - Theoretical study of unimolecular decomposition of catechol. AB - This study develops the reaction pathway map for the unimolecular decomposition of catechol, a model compound for various structural entities present in biomass, coal, and wood. Reaction rate constants at the high-pressure limit are calculated for the various possible initiation channels. It is found that catechol decomposition is initiated dominantly via hydroxyl H migration to a neighboring ortho carbon bearing an H atom. We identify the direct formation of o benzoquinone to be unimportant at all temperatures, consistent with the absence of this species from experimental measurements. At temperatures higher than 1000 K, water elimination through concerted expulsion of a hydroxyl OH together with an ortho H becomes the most significant channel. Rice-Ramsperger-Kassel-Marcus simulations are performed to establish the branching ratio between these two important channels as a function of temperature and pressure. All unimolecular routes to the reported major experimental products (CO, 1,3-C(4)H(6) and cyclo C(5)H(6)) are shown to incur large activation barriers. The results presented herein should be instrumental in gaining a better understanding of the decomposition behavior of catechol-related compounds. PMID- 20028003 TI - First-principles study of high-pressure behavior of solid beta-HMX. AB - A first-principles plane-wave method with an ultrasoft pseudopotential scheme in the framework of the generalized gradient approximation (GGA) was used to calculate the lattice parameters, bulk modulus and its pressure derivative, energy band structures, density of states, phonon density of states, thermodynamic properties, and absorption spectra of solid beta-octahydro-1,3,5,7 tetranitro-1,3,5,7-tetrazocine (beta-HMX). The current study is focused on the thermodynamics and electronic properties that were not reported previously. The bulk modulus and its pressure derivative are also consistent with experimental data and other theoretical results. From the results for the band gaps and density of states, it was found that beta-HMX has the tendency to become a semiconductor with increasing pressure. As the temperature increases, the heat capacity, enthalpy, product of temperature and entropy, and Debye temperature all increase, whereas the free energy decreases. The optical absorption coefficients shift to higher frequencies/energies with increasing pressure. The present study leads to a better understanding of how energetic materials respond to compression. PMID- 20028004 TI - Synthesis, radiolabeling, and biological evaluation of (R)- and (S)-2-amino-3 [(18)F]fluoro-2-methylpropanoic acid (FAMP) and (R)- and (S)-3-[(18)F]fluoro-2 methyl-2-N-(methylamino)propanoic acid (NMeFAMP) as potential PET radioligands for imaging brain tumors. AB - The non-natural amino acids (R)- and (S)-2-amino-3-fluoro-2-methylpropanoic acid 5 and (R)- and (S)-3-fluoro-2-methyl-2-N-(methylamino)propanoic acid 8 were synthesized in shorter reaction sequences than in the original report starting from enantiomerically pure (S)- and (R)-alpha-methyl-serine, respectively. The reaction sequence provided the cyclic sulfamidate precursors for radiosynthesis of (R)- and (S)-[(18)F]5 and (R)- and (S)-[(18)F]8 in fewer steps than in the original report. (R)- and (S)-[(18)F]5 and(R)- and (S)-[(18)F]8 were synthesized by no-carrier-added nucleophilic [(18)F]fluorination in 52-66% decay-corrected yields with radiochemical purity over 99%. The cell assays showed that all four compounds were substrates for amino acid transport and enter 9L rat gliosarcoma cells in vitro at least in part by system A amino acid transport. The biodistribution studies demonstrated that in vivo tumor to normal brain ratios for all compounds were high with ratios of 20:1 to115:1 in rats with intracranial 9L tumors. The (R)-enantiomers of [(18)F]5 and [(18)F]8 demonstrated higher tumor uptake in vivo compared to the (S)-enantiomers. PMID- 20028005 TI - A topological study of the decomposition of 6,7,8-trioxabicyclo[3.2.2]nonane induced by Fe(II): modeling the artemisinin reaction with heme. AB - We report a theoretical study on the electronic and topological aspects of the reaction of dihydrated Fe(OH)(2) with 6,7,8-trioxabicyclo[3.2.2]nonane, as a model for the reaction of heme with artemisinin. A comparison is made with the reaction of dihydrated ferrous hydroxide with O(2), as a model for the heme interaction with oxygen. We found that dihydrated Fe(OH)(2) reacts more efficiently with the artemisinin model than with O(2). This result suggests that artemisinin instead of molecular oxygen would interact with heme, disrupting its detoxification process by avoiding the initial heme to hemin oxidation, and killing in this way the malaria parasite. The ELF and AIM theories provide support for such a conclusion, which further clarifies our understanding on how artemisinin acts as an antimalarial agent. PMID- 20028006 TI - Interactions of phospholipid bilayers with several classes of amphiphilic alpha helical peptides: insights from coarse-grained molecular dynamics simulations. AB - In this article, we focus on several types of interactions between lipid membranes and alpha-helical peptides, based on the distribution of hydrophobic and hydrophilic residues along the helix. We employ a recently proposed coarse grained model MARTINI and test its ability to capture diverse types of behavior. MARTINI provides useful insights on the formation of barrel-stave and toroidal pores and on the relation between these two mechanisms. Amphipathic nonspanning peptides are also described with sufficient accuracy. The picture is not as clear for fusion and transmembrane peptides. For each class of peptides, we calculate the potential of mean force (PMF) for peptide translocation across the lipid bilayer and demonstrate that each class has a distinct shape of PMF. The reliability of these calculations, as well as wider implications of the results, is discussed. PMID- 20028007 TI - Soft glass rheology in liquid crystalline gels formed by a monodisperse dipeptide. AB - Thermal and extensive rheological characterization of a nematic liquid crystal gelated with a novel monodisperse dipeptide, also a liquid crystal, has been carried out. For certain concentrations, the calorimetric scans display a two peak profile across the chiral nematic-isotropic (N*-I) transition, a feature reminiscent of the random-dilution to random-field crossover observed in liquid crystal gels formed with aerosil particles. All samples show shear thinning behavior without a Newtonian plateau region at lower shear rates. Small deformation oscillatory data at lower frequencies exhibit a frequency dependence of the storage (G') and loss (G'') moduli that can be described by a weak power law, characteristic of soft glassy rheological systems. At higher frequencies, while lower concentration composites have a strong frequency dependence with a trend for possible crossover from viscoelastic solid to viscoelastic liquid behavior, the higher-concentration gels show frequency-independent rheograms of entirely elastic nature G' > G''. The plateau modulus of G' is described by a power-law with an exponent again common to soft materials, such as foams, slurries, etc. Other features which are a hallmark of such materials observed in the present study are: (i) above a critical strain, a strain softening of the moduli with a peak in the loss modulus, (ii) power-law variation of the storage modulus in the nonlinear viscoelastic regime, and (iii) absence of Cox-Merz superposition for the complex viscosity. An attractive feature of these gels is the fast recovery upon removal of large strain and qualitatively different temporal behavior of the recovery between the low and high concentration composites, with the latter indicating the presence of two characteristic time scales. PMID- 20028008 TI - Fluorescence enhancement in Langmuir-Blodgett films: role of amphiphile structure, orientation, and assembly. AB - Suppression of self-quenching of fluorescence emission in supramolecular systems is of fundamental interest in the design and development of novel optical materials. While a number of molecular design and assembly strategies have been formulated for the fabrication of molecular crystals with enhanced light emission, parallel explorations based on systematic approaches in the domain of molecular ultrathin films are rare. We have investigated Langmuir-Blodgett films of amphiphilic molecules bearing the same hemicyanine choromophore headgroup but with different possibilities of attachment of the octadecyl hydrocarbon chain, under different deposition conditions. Fluorescence spectroscopy studies indicate enhanced light emission in the derivative with a "tail-head-tail" structure. The observed trends are attributed to the amphiphile structure, nature of deposition, and the chromophore orientations revealed through polarized absorption spectra of the films. The study suggests a simple design strategy toward realization of molecular ultrathin films with enhanced light emission. PMID- 20028009 TI - Molecular dynamics and entropy effects in hydrogen-bonded supramolecular polymer N,N'-di(2-methyl-2-pentylheptyl)urea dissolved in nonpolar medium. AB - On the basis of the static dielectric permittivity temperature behavior of the supramolecular polymer formed by N,N'-di(2-methyl-2-pentylheptyl)urea (MPHU) in carbon tetrachloride diluted solution (10% in mole fraction) and that of neat acetonitrile, it was found that two liquids of the same macroscopic polarity (expressed by the same value of the dielectric permittivity approximately 35) exhibit an essential difference in the field-induced orientational entropy change. A much higher entropy effect is observed for the liquid composed of not too numerous but highly polar molecular entities (MPHU + CCl(4)) than for the liquid composed of numerous but less polar entities (neat acetonitrile). The analysis of the dielectric relaxation spectra of MPHU + CCl(4) solutions was performed with the Davidson-Cole (DC) model. It was found that the DC exponent beta changes its value in a quite important range of 0.4-0.8, depending on MPHU concentration and temperature. This reflects an important deviation of the relaxation mechanism occurring in the supramolecular system investigated from that corresponding to the normal Brownian rotational diffusion (beta = 1). PMID- 20028010 TI - Isolation and identification of chlorinated genistein from Actinoplanes sp. HBDN08 with antioxidant and antitumor activities. AB - A strain Actinoplanes sp. HBDN08 was screened by PCR-guided method using primers derived from conserved regions of halogenase genes. A new chlorinated isoflavone, 3',8-dichlorogenistein (1), along with 8-chlorogenistein (2) were isolated from the fermentation broth of Actinoplanes sp. HBDN08. Their structures were elucidated on the basis of extensive 1D and 2D NMR as well as HRESI-MS, ESI-MS, UV, and IR spectroscopic analyses. The origin of the two compounds was also investigated by high-performance liquid chromatography (HPLC) analysis. The results demonstrated that they were not biosynthesized but derived from the biotransformation of genistein by Actinoplanes sp. HBDN08. The antioxidant activities of the isolated compounds 1 and 2 were evaluated by using the lipid peroxidation assay. Their antitumor activities were calculated according to the inhibitory rate of cell proliferation against the human breast cancer cell line MDA-MB-231. The results indicated that compounds 1 (IC(50) = 5.2 microM) and 2 (IC(50) = 7.5 microM) showed stronger antioxidant activities than genistein (IC(50) = 13.6 microM). In comparison with the antitumor activities of genistein, those of compounds 1 and 2 increased 7.7- and 2.6-fold, respectively. These results suggest that the PCR-guided screening strategy is a rapid method for obtaining halometabolite-producing strains. Moreover, these results reveal that chlorination has significant effects on the bioactivities of genistein. This could be important information for studying the structure-activity relationships of genistein. PMID- 20028011 TI - Isolation and NMR characterization of fumonisin B2 and a new fumonisin B6 from Aspergillus niger. AB - A new fumonisin, fumonisin B(6) (1), has been isolated by cation-exchange and reverse-phase chromatography, together with fumonisin B(2) (2), from stationary cultures of the fungus Aspergillus niger NRRL 326. Analysis of mass spectrometric and NMR data determined that FB(6) is a positional isomer of FB(1) and iso-FB(1), having hydroxyl functions at C3, C4, and C5. Analysis of the NMR data for FB(2) showed very similar chemical shift values when compared to an authentic Fusarium FB(2) standard, strongly indicating identical molecules despite that an absolute stereochemical assignment of FB(2) from A. niger was not possible. PMID- 20028012 TI - Caffeoylquinic acids and flavonoids in the immature inflorescence of globe artichoke, wild cardoon, and cultivated cardoon. AB - The species Cynara cardunculus is consumed as part of the Mediterranean diet and consists of the globe artichoke [var. scolymus (L.) Fiori], the cultivated cardoon (var. altilis DC.), and the wild cardoon [var. sylvestris (Lamk) Fiori]. The objective of this study was to investigate, in immature inflorescences, the main flavonoids and phenolic acids (caffeoylquinic acids, apigenin, and luteolin derivatives) by HPLC/diode array detection/mass spectrometry. Apigenin derivatives represented the major class in all samples investigated, highest in cardoon forms. Caffeoylquinic acids and luteolin derivatives were observed in var. scolymus only. Data allowed discrimination of globe artichoke from the related species on the basis of the profile of compounds analyzed. Our results suggest the possible use of cultivated and wild cardoon as a source of phenolic acids and flavonoids and indicate that artichoke consumption is an excellent dietary source of apigenin and other flavones. PMID- 20028013 TI - Comparative safety evaluation of Chinese Pu-erh green tea extract and Pu-erh black tea extract in Wistar rats. AB - Pu-erh teas are believed to be beneficial beverages for health since they possess several pharmacological properties such as antioxidation, hypocholesterolemia, and antiobesity properties, but their potential toxicities when administered at a high dose as concentrated extracts have not been completely investigated. In this study, the chemical components in Pu-erh green tea and Pu-erh black tea were analyzed and compared, and the safety of tea extracts was evaluated in Wistar rats. The polysaccharide, tea pigment, and flavonoid levels were substantially increased in the Pu-erh black tea, while the polyphenol and free amino acid levels were higher in unfermented green tea. Low toxicities of Pu-erh green tea extract (GTE) were observed at doses of 2500 and 5000 mg/kg/day with a 28-day subacute study. Serum biochemical data including alanine aminotransferase increased to 5000 mg/kg/day GTE males, and creatinine (Cr) increased in all 5000 mg/kg/day GTE groups and 2500 mg/kg/day GTE males. Slight bile duct hyperplasia in the liver was also observed. The target organs of GTE were considered to be the liver and kidney. Comparatively, no adverse effects were observed in Pu-erh black tea extract (BTE)-treated rats. In conclusion, a dose of 1250 mg/kg/day for GTE and 5000 mg/kg/day for BTE following oral administration could be considered safe under the conditions of this study. PMID- 20028014 TI - Picrocrocin content and quality categories in different (345) worldwide samples of saffron ( Crocus sativus L.). AB - In this paper, 345 saffron samples were analyzed from different countries to study their picrocrocin content using different analytical techniques. The E1cm 1% of 257 nm results from all samples are inflated in comparison by the high performance liquid chromatography (HPLC) data, because of the interferences with the crocetin ester pool and especially with those with a lower trans/cis relation. A picrocrocin range update is proposed for International Organization for Standardization (ISO) 3632 normative because category III should be incremented up to 50 units, while category II should be incremented up to 60 units. More accurate data are achieved when the deltaEpic measurement is carried out. Consequently, improvements to the ISO method are suggested. Fourier transform (FT)-near-infrared spectrometry analysis has also been carried out, showing excellent results from the calibration with HPLC data. This spectrophotometric technique could be used by saffron enterprises to obtain quick and more accurate data for picrocrocin determination. PMID- 20028015 TI - Development and validation of an indirect competitive enzyme linked-immunosorbent assay for the determination of potentially allergenic sesame (Sesamum indicum) in food. AB - This study was designed to develop an indirect competitive enzyme linked immunosorbent assay (ELISA) to detect traces of sesame in food. Antibodies against sesame were prepared by immunizing a hen with a protein extract of white, peeled sesame. The ELISA did not show any cross-reactivity with 12 of 13 food ingredients tested, only for chocolate was a low cross-reactivity of 0.7% observed. To eliminate matrix effects, sesame protein standard solutions were prepared by diluting the sesame extract with blank food matrix (1:20 diluted with PBS). Recovery of sesame protein in food samples (crisp toasts, snacks, and rolls) spiked with different sesame protein concentrations ranged from 85% to 120%, with the exception of multigrain crisp toast, resulting in too high recoveries (117%-160%) and whole grain bread, yielding too low recoveries (70% 85%). In crisp bread, cracker, cereals, and snacks the limit of detection (LOD) was found to be 5 microg of sesame protein/g of food, in fresh breads and rolls, the LOD was 11 microg of sesame protein/g of food. PMID- 20028016 TI - Acids, sugars, and sugar alcohols in Chinese hawthorn (Crataegus spp.) fruits. AB - Acids, sugars, and sugar alcohols in the fruits of 22 cultivars/origins of three species of hawthorn (Crataegus spp.) were analyzed by gas chromatography and mass spectrometry. Citric acid (2.0-8.4 g/100 g dry mass [DM]), quinic acid (0.5-5.6 g/100 g DM), malic acid (0.3-1.1 g/100 g DM), fructose (5.5-18.4 g/100 g DM), glucose (5.3-16.6 g/100 g DM), sorbitol (3.0-15.7 g/100 g DM), and myo-inositol (0.1-0.3 g/100 g DM) were found in all the samples. Sucrose was present only in C. scabrifolia and three cultivars of C. pinnatifida var. major. C. scabrifolia differed from other species by its high content of quinic acid. The cultivars of C. pinnatifida var. major and C. brettschneideri had a higher content of total sugars and a higher sugar/acid ratio than the natural origins of C. pinnatifida and C. scabrifolia (P < 0.05). The hawthorn samples analyzed fell into two groups rich in sugars and acids respectively. This is the first report of the profiles of sugars and sugar alcohols and the occurrence of quinic acid in hawthorn fruits. PMID- 20028017 TI - Optimization of pressurized liquid extraction of carotenoids and chlorophylls from Chlorella vulgaris. AB - Pressurized liquid extraction (PLE) was applied to the extraction of carotenoids and chlorophylls from the green microalga Chlorella vulgaris. Four extraction techniques such as maceration (MAC), Soxhlet extraction (SOX), ultrasound assisted extraction (UAE), and PLE were compared, and both the extraction temperature (50, 105, and 160 degrees C) and the extraction time (8, 19, and 30 min), which are the two main factors for PLE, were optimized with a central composite design to obtain the highest extraction efficiency. The extraction solvent (90% ethanol/water) could adequately extract the functional components from C. vulgaris. PLE showed higher extraction efficiencies than MAC, SOX, and UAE. Temperature was the key parameter having the strongest influence on the extraction of carotenoids and chlorophylls from chlorella. In addition, high heat treatment (>110 degrees C) by PLE minimized the formation of pheophorbide a, a harmful chlorophyll derivative. These results indicate that PLE may be a useful extraction method for the simultaneous extraction of carotenoids and chlorophylls from C. vulgaris. PMID- 20028018 TI - Pesticide multiresidue analysis in cereal grains using modified QuEChERS method combined with automated direct sample introduction GC-TOFMS and UPLC-MS/MS techniques. AB - The QuEChERS (quick, easy, cheap, effective, rugged, and safe) sample preparation method was modified to accommodate various cereal grain matrices (corn, oat, rice, and wheat) and provide good analytical results (recoveries in the range of 70-120% and RSDs <20%) for the majority of the target pesticides (about 180 analytes). The method consists of a 1 h shaking of a milled sample (2.5-5 g) in 20 mL of 1:1 (v/v) water/acetonitrile (or 25 mL of 1.5:1 water/acetonitrile in the case of rice) to provide simultaneous matrix swelling and analyte extraction. Then, a MgSO(4)/NaCl salt mixture (4:1, w/w) is added to the extract to induce phase separation and force the pesticides into the upper acetonitrile layer, a 1 mL aliquot of which is subsequently cleaned up using dispersive solid phase extraction with 150 mg of PSA, 50 mg of C(18), and 150 mg of MgSO(4). GC-amenable pesticides were analyzed using gas chromatography combined with time-of-flight mass spectrometry (GC-TOFMS) and the automated direct sample introduction technique for a large volume injection of the extracts. Ultraperformance liquid chromatography coupled to triple-quadrupole tandem mass spectrometry (UPLC-MS/MS) was employed for the analysis of LC-amenable pesticides. This method was implemented in a routine laboratory, providing about 3-fold increased sample throughput, 40-50% reduction in the cost of disposable materials and in the operation costs, 1:100 solvent waste reduction, and increased scope of the analysis versus the traditional approach based on the Luke method. PMID- 20028019 TI - Revalorization of a two-phase olive mill waste extract into a micronutrient fertilizer. AB - Micronutrient deficiencies in plants may be treated using metal complexes. A modified two-phase olive mill waste (OMWm) was characterized using FTIR spectroscopy. A study was also made of micronutrient (Fe, Zn, Mn and Cu) complexation and the stability of complexes. An evaluation was then made of the effectiveness of Fe(III)OMWm to supply Fe to soybean (Glycine max. cv Stine 0480) chlorotic plants through nutrient solution and foliar application. The OMWm presented structural similarities with the fulvic fraction of raw OMW containing abundant phenolic-hydroxyl and carboxyl groups able to form complexes. The OMWm could complex Fe, Zn, Mn and Cu, although the stabilities of the complexes under agronomic conditions were low. In dealing with Fe chlorosis, Fe(III)-OMWm improved the Fe nutritional status of soybean chlorotic plants when applied to the nutrient solution, while only regreening of leaves was observed in foliar applications. Hence, OMWm complexes constitute a promising eco-compatible and cheap alternative to synthetic chelates in dealing with micronutrient deficiencies when applied foliarly or to the nutrient solution, although further research is necessary to improve the stability and effectiveness of the complexes. PMID- 20028020 TI - Metal- and anion-binding supramolecular gels. PMID- 20028021 TI - Tunable detection sensitivity of opiates in urine via a label-free porous silicon competitive inhibition immunosensor. AB - Currently, there is need for laboratory-based high-throughput and reliable point of-care drug screening methodologies. We demonstrate here a chip-based label-free porous silicon (PSi) photonic sensor for detecting opiates in urine. This technique provides a cost-effective alternative to conventional labeled drug screening immunoassays with potential for translation to multiplexed analysis. Important effects of surface chemistry and competitive binding assay protocol on the sensitivity of opiate detection are revealed. Capability to tune sensitivity and detection range over approximately 3 orders of magnitude (18.0 nM to 10.8 muM) was achieved by varying the applied urine specimen volume (100-5 muL), which results in systematic shifts in the competitive binding response curve. A detection range (0.36-4.02 muM) of morphine in urine (15 muL) was designed to span the current positive cutoff value (1.05 muM morphine) in medical opiate urine screening. Desirable high cross-reactivity to oxycodone, in addition to other common opiates, morphine, morphine-3-glucuronide, 6-acetyl morphine, demonstrates an advantage over current commercial screening assays, while low interference with cocaine metabolite was maintained. This study uniquely displays PSi sensor technology as an inexpensive, rapid, and reliable drug screening technology. Furthermore, the versatile surface chemistry developed can be implemented on a range of solid-supported sensors to conduct competitive inhibition assays. PMID- 20028022 TI - Cleavable linker for photo-cross-linked small-molecule affinity matrix. AB - The introduction of a cleavable site in a photoactivatable linker, which is used to immobilize small molecules on an affinity matrix via a site-nonselective carbene addition/insertion reaction, makes it possible to verify the presence of the immobilized small molecule on the affinity matrix. It also permits the efficient detection of proteins covalently bound to the immobilized small molecule. PMID- 20028023 TI - Isotopomics: a top-down systems biology approach for understanding dynamic metabolism in rats using [1,2-(13)C(2)] acetate. AB - Isotope labeled tracers are commonly used to quantify the turnover rates of various metabolic intermediates and yield information regarding physiological regulation. Studies often only consider either one nutritional state (fasted or fed) and/or one question (e.g., measure of lipid or protein turnover). In this article, we consider a novel application combining the global approach of metabonomics with widespread stable isotope labeling as a way of being able to map metabolism in open mammalian systems, an approach we call "isotopomics". A total of 45 15-week-old male Zucker rats were administrated different amounts (from 0.5 to 8 mmol/kg) of sodium [1,2-(13)C(2)] acetate. Plasma samples taken at 1, 4, and 24 h were analyzed with (13)C nuclear magnetic resonance (NMR) and gas chromatography/mass spectrometry (GC/MS) to measure (13)C isotopic enrichment of 39 plasma metabolites across a wide range of compound classes (amino acids, short chain fatty acids, lactate, glucose, and free fatty acids). Isotopic enrichment from 0.1-7.1 mole percent excess (MPE) for the highest dose could be reliably measured in 16 metabolites, and the kinetics of their (13)C isotopic enrichment are reported. Clustering metabolites based on (13)C kinetic curves enabled highlighting of time dependent patterns of (13)C distribution through the key metabolic pathways. These kinetic and quantitative data were reported into a biochemical map. This type of isotopomic approach for mapping dynamic metabolism in an open system has great potential for advancing our mechanistic knowledge of how different interventions and diseases can impact the metabolic response of animals and humans. PMID- 20028024 TI - Activity-based proteome profiling of potential cellular targets of Orlistat--an FDA-approved drug with anti-tumor activities. AB - Orlistat, or tetrahydrolipstatin (THL), is an FDA-approved antiobesity drug with potential antitumor activities. Cellular off-targets and potential side effects of Orlistat in cancer therapies, however, have not been extensively explored thus far. In this study, we report the total of synthesis of THL-like protein-reactive probes, in which extremely conservative modifications (i.e., an alkyne handle) were introduced in the parental THL structure to maintain the native biological properties of Orlistat, while providing the necessary functionality for target identification via the bio-orthogonal click chemistry. With these natural productlike, cell-permeable probes, we were able to demonstrate, for the first time, this chemical proteomic approach is suitable for the identification of previously unknown cellular targets of Orlistat. In addition to the expected fatty acid synthase (FAS), we identified a total of eight new targets, some of which were further validated by experiments including Western blotting, recombinant protein expression, and site-directed mutagenesis. Our findings have important implications in the consideration of Orlistat as a potential anticancer drug at its early stages of development for cancer therapy. Our strategy should be broadly useful for off-target identification against quite a number of existing drugs and/or candidates, which are also covalent modifiers of their biological targets. PMID- 20028025 TI - C-H activation for the construction of C-B bonds. PMID- 20028027 TI - Nakijiquinones J--R, sesquiterpenoid quinones with an amine residue from okinawan marine sponges. AB - Nine new sesquiterpenoid quinones, nakijiquinones J-R (1-9), have been isolated from three collections of Okinawan marine sponges of the family Spongiidae, and the structures and configurations were elucidated from the spectroscopic data and chemical correlations. Nakijiquinones J-L (1-3), M and N (4 and 5, respectively), O (6), P and Q (7 and 8, respectively), and R (9) are new sesquiterpenoid quinones possessing (S)-2-methylbutylamine, isopentylamine, isobutylamine, phenethylamine, and taurine residues, respectively, attached to each quinone ring. PMID- 20028026 TI - An efficient synthetic strategy for obtaining 4-methoxy carbon isotope labeled combretastatin A-4 phosphate and other Z-combretastatins. AB - Human cancer and other clinical trials under development employing combretastatin A-4 phosphate (1b, CA4P) should benefit from the availability of a [(11)C] labeled derivative for positron emission tomography (PET). In order to obtain a suitable precursor for addition of a [(11)C]methyl group at the penultimate step, several new synthetic pathways to CA4P were evaluated. Geometrical isomerization (Z to E) proved to be a challenge, but it was overcome by development of a new CA4P synthesis suitable for 4-methoxy isotope labeling. PMID- 20028028 TI - Resonant plasmon nanofocusing by closed tapered gaps. AB - We study radiation nanofocusing by closed tapered gaps, i.e. metal V-grooves, under normal illumination, and discover that the local field inside a groove can be resonantly enhanced due to interference of counter-propagating gap plasmons. Considering V-grooves milled in gold, we analyze this phenomenon theoretically, deriving an analytic expression for the resonance condition and predicting more than 550-fold intensity enhancements at resonance, and observe it experimentally with two-photon photoluminescence microscopy, demonstrating more than 100-fold intensity enhancements. PMID- 20028029 TI - An antimycobacterial cyclodepsipeptide from the entomopathogenic fungus Ophiocordyceps communis BCC 16475. AB - A novel cyclodepsipeptide, cordycommunin (1), and two dihydroisocoumarins (2 and 3) were isolated from the insect pathogenic fungus Ophiocordyceps communis BCC 16475. The absolute configurations of the amino acid residues of 1 were addressed by application of Marfey's method. Cordycommunin (1) showed growth inhibition of Mycobacterium tuberculosis H37Ra with an MIC value of 15 microM. This compound also exhibited weak cytotoxicity to KB cells with an IC50 of 45 microM, while it was inactive against BC, NCI-H187, and Vero cell lines at a concentration of 88 microM (50 microg/mL). PMID- 20028030 TI - Neocomplanines A and B, a complanine family isolated from the marine fireworm. AB - Two new members of the complanine family, neocomplanines A (2) and B (3), were isolated as minor components of the methanolic extract of the "marine fireworm" Eurythoe complanata. The structures of the neocomplanines were revealed spectroscopically to be a trimethylammonium unit with a saturated carbon chain. The planar structures of neocomplanines A and B were confirmed successfully through total syntheses that used epichlorohydrin as a starting material. The neocomplanines show inflammatory activity and also enhanced PKC activity in combination with TPA in the presence of Ca(2+) in vitro; both are similar to the effects of complanine (1). The molecular mechanism of the effects of complanine related compounds is discussed. PMID- 20028032 TI - Triarylamines designed to form molecular glasses. Derivatives of tris(p-terphenyl 4-yl)amine with multiple contiguous phenyl substituents. AB - The principles of crystal engineering can be used in a contrary way to help devise molecules that resist crystallization and form long-lived glasses. This can be achieved by making structural changes that thwart established patterns of crystallization. In using this strategy to block the crystallization of triarylamines, we have found that the introduction of methylpentaphenyl groups is particularly effective, presumably because they inhibit efficient molecular packing and normal intermolecular interactions. PMID- 20028033 TI - Syntheses and crystal structures of benzohexathia[7]helicene and naphthalene cored double helicene. AB - Bis(trimethylsilyl)benzohexathia[7]helicene 1, naphthalene cored double helicene 2 (the fused dimer of 1), and a novel ten-membered cyclic diketone with four moieties of dithieno[2,3-b:3',2'-d]thiophene (3) were efficiently synthesized. Their crystal structures were determined with single-crystal X-ray analysis. In their crystal packings, they all show multiple short contacts including intermolecular pi...pi, pi...S, and S...S interactions. UV/vis spectra indicate that significant pi-electron delocalization existed in 1, 2, and 3. PMID- 20028036 TI - Potential of geoengineering highly uncertain. PMID- 20028037 TI - Particulate matter in the air and its origins in coal-burning regions. PMID- 20028034 TI - Residues essential for plasminogen binding by the cation-independent mannose 6 phosphate receptor. AB - The 300 kDa cation-independent mannose 6-phosphate receptor (CI-MPR) is a multifunctional protein that binds diverse intracellular and extracellular ligands with high affinity. The CI-MPR is a receptor for plasminogen, and this interaction can be inhibited by lysine analogues. To characterize the molecular basis for this interaction, surface plasmon resonance (SPR) analyses were performed using truncated forms of the CI-MPR and plasminogen. The results show that the N-terminal region of the CI-MPR containing domains 1 and 2, but not domain 1 alone, of the receptor's 15-domain extracytoplasmic region binds plasminogen (K(d) = 5 +/- 1 nM) with an affinity similar to that of the full length receptor (K(d) = 20 +/- 6 nM). In addition to its C-terminal serine protease domain, plasminogen contains lysine binding sites (LBS), which are located within each of its five kringle domains, except kringle 3. We show that kringles 1-4, but not kringles 1-3, bind the CI-MPR, indicating an essential role for the LBS in kringle 4 of plasminogen. To identify the lysine residue(s) of the CI-MPR that serve(s) as an essential determinant for recognition by the LBS of plasminogen, site-directed mutagenesis studies were carried out using a construct encoding the N-terminal three domains of the CI-MPR (Dom1-3His) which contains both a mannose 6-phosphate (Man-6-P) and plasminogen binding site. The results demonstrate two lysine residues (Lys53 located in domain 1 and Lys125 located in the loop connecting domains 1 and 2) of the CI-MPR are key determinants for plasminogen binding but are not required for Man-6-P binding. PMID- 20028038 TI - UN update: climate change hitting sooner and stronger. PMID- 20028039 TI - Contaminants reveal tuna's origins. PMID- 20028040 TI - Importance of considering the framework principles in risk assessment for metals. AB - The recent EPA Framework for Metals Risk Assessment provides the opportunity for contextual risk assessment for sites impacted by metals (such as the depicted Dauntless Mine in Colorado). PMID- 20028041 TI - Emerging opportunities in management of selenium contamination. AB - Integrating the chemistry of selenium with its biology and ecotoxicology gives indications on how to regulate its environmental levels. PMID- 20028042 TI - The arsenic hyperaccumulator fern Pteris vittata L. AB - Arsenic (As) contaminated soils and waters are becoming major global environmental and human health risks. The identification of natural hyperaccumulators of As opens the door for phytoremediation of the arsenic contaminant. Pteris vittata is the first identified naturally evolving As hyperaccumulator. More than a decade after its discovery, we have made great progress in understanding the uptake, transport, and detoxification of As in the fern. The molecular mechanisms controlling As accumulation in P. vittata are now beginning to be recognized. In this review, we will try to summarize what we have learned about this As accumulator, with particular emphasis on the current knowledge of the physiological and molecular mechanisms of arsenic phytoremediation. We also discuss the potential strategies to further enhance phytoextraction abilities of P. vittata. PMID- 20028043 TI - Review of the integrated process for the production of grass biomethane. AB - Production of grass biomethane is an integrated process which involves numerous stages with numerous permutations. The grass grown can be of numerous species, and it can involve numerous cuts. The lignocellulosic content of grass increases with maturity of grass; the first cut offers more methane potential than the later cuts. Water-soluble carbohydrates (WSC) are higher (and as such methane potential is higher) for grass cut in the afternoon as opposed to that cut in the morning. The method of ensiling has a significant effect on the dry solids content of the grass silage. Pit or clamp silage in southern Germany and Austria has a solids content of about 40%; warm dry summers allow wilting of the grass before ensiling. In temperate oceanic climates like Ireland, pit silage has a solids content of about 21% while bale silage has a solids content of 32%. Biogas production is related to mass of volatile solids rather than mass of silage; typically one ton of volatile solid produces 300 m(3) of methane. The dry solids content of the silage has a significant impact on the biodigester configuration. Silage with a high solids content would lend itself to a two-stage process; a leach bed where volatile solids are converted to a leachate high in chemical oxygen demand (COD), followed by an upflow anaerobic sludge blanket where the COD can be converted efficiently to CH(4). Alternative configurations include wet continuous processes such as the ubiquitous continuously stirred tank reactor; this necessitates significant dilution of the feedstock to effect a solids content of 12%. Various pretreatment methods may be employed especially if the hydrolytic step is separated from the methanogenic step. Size reduction, thermal, and enzymatic methodologies are used. Good digester design is to seek to emulate the cow, thus rumen fluid offers great potential for hydrolysis. PMID- 20028044 TI - Categorization of Scope 3 emissions for streamlined enterprise carbon footprinting. AB - Many organizations look to carbon footprint protocols for guidance on measuring their greenhouse gas emissions, or carbon footprint. Existing protocols generally require estimation of direct emissions (Scope 1) and emissions from direct purchases of energy (Scope 2), but focus less on indirect emissions upstream and downstream of the supply chain (optional Scope 3). Because on average more than 75% of an industry sector's carbon footprint is attributed to Scope 3 sources, better knowledge of Scope 3 footprints can help organizations pursue emissions mitigation projects not just within their own plants but also across their supply chain. In this work, Scope 3 footprints of U.S. economic sectors are categorized using an Economic Input-Output Life Cycle Assessment (EIO-LCA) model to identify upstream emission sources that are likely to contribute significantly to different sectors' footprints. The portions of the upstream footprint captured by the sector's top-10 upstream suppliers are estimated at 3 different levels of specificity: general economy-wide, industry specific, and sector specific. The results show that enterprises can capture a large portion of their total upstream carbon footprint by collecting full emissions information from only a handful of direct suppliers, and Scope 3 footprint capture rates can be improved considerably by sector-specific categorization. Employee commuting and air transportation may be more important (7%-30%) for the services industries, but should not be a focus of detailed Scope 3 footprint estimates for the manufacturing industries (<1% of the total analyzed footprint). Protocol organizations should actively make more specific Scope 3 guidelines available for their constituents by developing sector-specific categorizations for as many sectors as they feasibly can and create broader industry-specific protocols for others. PMID- 20028045 TI - Nitrogen dioxide and ozone as factors in the availability of lead from lead-based paints. AB - Lead-based paint remains a pervasive problem in U.S. cities, and an increasing problem in the developing world where it is still manufactured and used. Little attention has focused on the factors that increase the release of lead pigment granules from painted surfaces. Nitrogen dioxide (NO(2)) and ozone (O(3)) from transportation emissions in urban environments have the potential to react with and remove polymeric binders in paint, making pigment granules more available for subsequent transfer to hands on contact, or deposition in housedust. Here we show that exposure to NO(2) and O(3) increased the lead in wipe samples of stainless steel surfaces painted with alkyd low gloss solvent lead-based paint by 296% +/- 101 (or 0.24 microg/cm(2)) and 37% +/- 21 (or 0.025 microg/cm(2)), respectively, with corresponding changes in surface morphology indicated by reflectometry and scanning electron microscopy. Lead release from unexposed low gloss acrylic household paints was 40 times greater than comparable solvent based paints. Given that lead-based paint is still manufactured and used in many urban areas of the developing world where O(3) concentrations currently exceed historic U.S. concentrations, the interaction of air pollution with lead painted indoor surfaces may pose greater exposure risks for lead poisoning in children than previously anticipated. PMID- 20028046 TI - Atlantic bluefin tuna (Thunnus thynnus) population dynamics delineated by organochlorine tracers. AB - Atlantic bluefin tuna (ABFT) are highly valued and heavily exploited, and critical uncertainties regarding their population structure hinder effective management. Evidence supports the existence of two breeding populations of ABFT; a western population in the Gulf of Mexico and an eastern population in the Mediterranean Sea; both of which migrate and mix in the North Atlantic. Conventional tagging studies suggest low rates of trans-Atlantic migrations; however, electronic tagging and stable isotopes in otoliths indicate stock mixing up to 57% between management zones delineated by 45 degrees W longitude. Here we show that organochlorine pesticides and polychlorinated biphenyls (PCBs) can be used as tracers of bluefin tuna foraging grounds in the North Atlantic and confirm that stock mixing of juvenile tuna within the U.S. Mid Atlantic Bight is indeed high (33-83% eastern origin), and is likely spatially and temporally variable. We further demonstrate that >10% of the Mediterranean population is migratory, that young bluefin tuna migrate from the Mediterranean to western Atlantic foraging grounds as early as age 1, and then return to the Mediterranean Sea as young as age 5, presumably to breed. The tracer method described here provides a novel means for distinguishing bluefin tuna populations and ontogenetic shifts in migration in the North Atlantic. PMID- 20028047 TI - Uranium in framboidal pyrite from a naturally bioreduced alluvial sediment. AB - Samples of a naturally bioreduced, U-contaminated alluvial sediment were characterized with various microscopic and spectroscopic techniques and wet chemical extraction methods. The objective was to investigate U association and interaction with minerals of the sediment. Bioreduced sediment comprises approximately 10% of an alluvial aquifer adjacent to the Colorado River, in Rifle, CO, that was the site of a former U milling operation. Past and ongoing research has demonstrated that bioreduced sediment is elevated in solid associated U, total organic carbon, and acid-volatile sulfide, and depleted in bioavailable Fe(III) confirming that sulfate and Fe(III) reduction have occurred naturally in the sediment. SEM/EDS analyses demonstrated that framboidal pyrites (FeS(2)) of different sizes ( approximately 10-20 microm in diameter), and of various microcrystal morphology, degree of surface weathering, and internal porosity were abundant in the <53 microm fraction (silt + clay) of the sediment and absent in adjacent sediments that were not bioreduced. SEM-EMPA, XRF, EXAFS, and XANES measurements showed elevated U was present in framboidal pyrite as both U(VI) and U(IV). This result indicates that U may be sequestered in situ under conditions of microbially driven sulfate reduction and pyrite formation. Conversely, such pyrites in alluvial sediments provide a long-term source of U under conditions of slow oxidation, contributing to the persistence of U of some U plumes. These results may also help in developing remedial measures for U contaminated aquifers. PMID- 20028048 TI - Elevated concentrations of methyl mercury in streams after forest clear-cut: a consequence of mobilization from soil or new methylation? AB - Concentrations of inorganic, mercuric mercury (Hg(II)), methyl mercury (MeHg) and ancillary chemistry measured in first-order streams draining 0-4 (N = 20) and 4 10 (N = 27) year-old clear-cuts of former Norway Spruce Picea abies (Karst.) forest stands were compared with concentrations in streams draining >70 year-old Norway Spruce reference stands (N = 10). Concentrations of MeHg, and ratios of MeHg TOC(-1) and Hg(II) TOC(-1), were significantly (p < 0.01) elevated in 0-4 year-old clear-cuts, as compared to references. The only ancillary variable showing a significant elevation for 0-4 year-old clear-cuts was Mn (p < 0.02). The 4-10 year-old clear-cuts showed intermediate concentrations with nonsignificant differences as compared to references. pH, nitrate, sulfate, Ca, Fe, TOC, TON, and the aromaticity of TOC (SUVA(254 nm)) showed nonsignificant differences between clear-cut age classes and references. Assuming that MeHg and Hg(II) are mobilized from soil to stream to a similar relative extent as a consequence of clear-cutting, a calculation showed that (1)/(6) of the elevated MeHg concentration was due to enhanced mobilization from soil and (5)/(6) was due to new methylation of Hg(II) 0-4 years after clear-cut. New methylation after clear-cut is suggested to be stimulated by an increased availability of electron donors for methylating bacteria, as a consequence of degradation of logging residue ("slash") and soil organic matter. A subdivision of sites situated above and below the highest postglacial coastline (HC) revealed a significant elevation of MeHg, MeHg TOC(-1) and Hg(II) TOC(-1) (p < 0.05) beyond their references in 0 4 year-old clear-cuts above (but not below) the HC. This suggests that postglacial deposits of FeS(s) and FeS(2)(s) were not an important factor for elevation of MeHg after clear-cut. PMID- 20028049 TI - Gas/particle partitioning behavior of perfluorocarboxylic acids with terrestrial aerosols. AB - Experimentally determined gas/particle partitioning constants, K(ip), using inverse gas chromatography (IGC) are presented for perfluorocarboxylic acids (PFCAs), covering a diverse set of terrestrial aerosols over an ambient range of relative humidity (RH) and temperature. The results are compared to estimated K(ip) values using a recently developed model that has been validated for diverse neutral and ionizable organic compounds. The modeling results consistently underestimate the experimental results. This is likely due to additional partition mechanisms unique for surfactants not being accounted for in the model, namely aggregate formation and water surface adsorption. These processes likely also biased the IGC K(ip) measurements compared to ambient PFCA concentrations. Nevertheless, both the experimental and modeling results indicate that partitioning to terrestrial particles in ambient atmospheres is negligible, though sorption to condensed water can be substantial. This favors rain sequestration as a more important atmospheric removal mechanism than dry particle sequestration. PFCAs found on particle filters during ambient sampling are thus accountable to vapor-phase PFCAs or aqueous-phase PFCAs sorbing directly to the filters, or the trapping of perfluorocarboxylate-salt particles. Further work on understanding the partitioning and speciation of PFCAs in atmospheric water droplets is needed to further quantify and understand their atmospheric behavior. To aid in this, a general RH dependent K(ip) model for surfactants is presented. PMID- 20028050 TI - Kinetic controls on the complexation between mercury and dissolved organic matter in a contaminated environment. AB - The interaction of mercury (Hg) with dissolved natural organic matter (NOM) under equilibrium conditions is the focus of many studies but the kinetic controls on Hg-NOM complexation in aquatic systems have often been overlooked. We examined the rates of Hg-NOM complexation both in a contaminated Upper East Fork Poplar Creek (UEFPC) in Oak Ridge, Tennessee, and in controlled laboratory experiments using reducible Hg (Hg(R)) measurements and C(18) solid phase extraction techniques. Of the filterable Hg at the headwaters of UEFPC, >90% was present as Hg(R) and this fraction decreased downstream but remained >29% of the filterable Hg at all sites. The presence of higher Hg(R) concentrations than would be predicted under equilibrium conditions in UEFPC and in experiments with a NOM isolate suggests that kinetic reactions are controlling the complexation between Hg and NOM. The slow formation of Hg-NOM complexes is attributed to competitive ligand exchange among various moieties and functional groups in NOM with a range of binding strengths and configurations. This study demonstrates the need to consider the effects of Hg-NOM complexation kinetics on processes such as Hg methylation and solid phase partitioning. PMID- 20028051 TI - Yields of glyoxal and ring-cleavage co-products from the OH radical-initiated reactions of naphthalene and selected alkylnaphthalenes. AB - Naphthalene and alkylnaphthalenes are the most abundant polycyclic aromatic hydrocarbons present in ambient air and are transformed mainly by chemical reaction with hydroxyl (OH) radicals during daylight hours. To better understand the reaction mechanisms, we have quantified glyoxal from the OH radical-initiated reactions of naphthalene, 1-methylnaphthalene, 1,4-dimethylnaphthalene, acenaphthene, and acenaphthylene as a function of the NO(2) concentration and, for the naphthalene reaction, also in the absence of NO(2). Glyoxal was formed as a first-generation product from the naphthalene, 1-methylnaphthalene, 1,4 dimethylnaphthalene, and acenaphthene reactions, and its yields were independent of the NO(2) concentration over the ranges employed, being 5% in the presence of NO(2) and 3% in the absence of NO(2) from naphthalene; approximately 3% from 1 methylnaphthalene; approximately 2% from 1,4-dimethylnaphthalene; approximately 10-15% from acenaphthene; and <2% from acenaphthylene. Second-generation formation of glyoxal was evident in the 1-methylnaphthalene, 1,4 dimethylnaphthalene, and acenaphthene reactions. For the naphthalene reaction, our results suggest that the reactions of the OH-naphthalene adducts with NO(2) and O(2) both lead to glyoxal formation in similar yield. Simultaneous measurements of phthaldialdehyde from naphthalene, 2-acetylbenzaldehyde from 1 methylnaphthalene, and 1,2-diacetylbenzene from 1,4-dimethylnaphthalene suggest that these C(n-2)-dicarbonyls are coproducts to glyoxal. PMID- 20028052 TI - Free water content and sludge retention time: impact on oxygen transfer in activated sludge. AB - To investigate the effect of sludge retention time (SRT) and the concentration of mixed liquid volatile suspended solids (MLVSS) on oxygen transfer in activated sludge, we studied mass transfer coefficients (k(L)a) in a pilot-scale membrane bioreactor, which treated synthetic greywater. Additionally, experiments with iron hydroxide flocs were performed to examine the role of free water content in oxygen transfer. The results demonstrate that the alpha-factor is reduced when free water content decreases and floc volume increases. Because the MLVSS concentration in activated sludge mainly regulates floc volume, a reduction of oxygen transfer with increasing MLVSS concentration was observed. If the floc volume remains unchanged, oxygen transfer increases with increasing SRT. PMID- 20028053 TI - Isomer-specific biotransformation rates of a perfluorooctane sulfonate (PFOS) precursor by cytochrome P450 isozymes and human liver microsomes. AB - The exposure sources of perfluorooctane sulfonate (PFOS) in humans and wildlife are not well characterized. Human biomonitoring data show that PFOS profiles may consist of up to approximately 50% branched isomers, despite the fact that historical direct manufacturing of PFOS generally resulted in products containing no more than approximately 30% branched isomers. These observations cannot be explained based on what is known about the pharmacokinetics of branched PFOS isomers; thus, here we examined the relative isomer-specific biotransformation rates of a model PFOS-precursor (N-ethylperfluorooctane sulfonamide, NEtFOSA) with human microsomes and recombinant human cytochrome P450s (CYPs) 2C9 and 2C19. Using solid phase microextraction-gas chromatography-electron capture detection to monitor NEtFOSA disappearance, and liquid chromatography-tandem mass spectrometry to monitor product formation, we showed that, in general, human microsomes and CYP isozymes transformed the branched isomers more rapidly than linear NEtFOSA. Among branched isomers, perfluoroalkyl branching geometry significantly influenced the rate of biotransformation. As a result, PFOS isomer patterns in biota exposed predominantly to precursors could be much different than expected from the isomer pattern of the precursor. While these data are suggestive that the relatively high abundance of branched PFOS isomers present in some humans, or wildlife, may be explained by substantial exposure to PFOS precursors, in vivo studies with other relevant PFOS-precursors are warranted to validate this as a biomarker of exposure source. PMID- 20028054 TI - Hemispheres-in-cell geometry to predict colloid deposition in porous media. AB - A "hemispheres-in-cell" geometry is provided for prediction of colloid retention during transport in porous media. This new geometry preserves the utilities provided in the Happel sphere-in-cell geometry; namely, the ability to predict deposition for a range of porosities, and representation of the influence of neighboring collectors on the fluid flow field. The new geometry, which includes grain to grain contact, is justified by the eventual goal of predicting colloid deposition in the presence of energy barriers, which has been shown in previous literature to involve deposition within grain to grain contacts for colloid:collector ratios greater than approximately 0.005. In order to serve as a platform for predicting deposition in the presence of energy barriers, the model must be shown capable of quantitatively predicting deposition in the absence of energy barriers, which is a requirement that was not met by previous grain to grain contact geometries. This paper describes development of the fluid flow field and particle trajectory simulations for the hemispheres-in-cell geometry in the absence of energy barriers, and demonstrates that the resulting simulations compare favorably to existing models and experiments. A correlation equation for predicting collector efficiencies in the hemispheres-in-cell model in the absence of energy barriers was developed via regression of numerical results to dimensionless parameters. PMID- 20028055 TI - Speciated VOC emission inventory and spatial patterns of ozone formation potential in the Pearl River Delta, China. AB - The Pearl River Delta region (PRD) of China has long suffered from severe ground level ozone pollution. Knowledge of the sources of volatile organic compounds (VOCs) is essential for ozone chemistry. In this work, a speciated VOC emission inventory was established on the basis of updated emissions and local VOC source profiles. The top 10 species, in terms of ozone formation potentials (OFPs), consisted of isoprene, mp-xylene, toluene, ethylene, propene, o-xylene, 1,2,4 trimethylbenzene, 2-methyl-2-butene, 1-butene, and alpha-pinene. These species contributed only 35.9% to VOCs emissions but accounted for 64.1% of the OFP in the region. The spatial patterns of the VOC source inventory agreed well with city-based source apportionment results, especially for vehicle emissions and industry plus VOC product-related emissions. Mapping of the OFPs and measured ozone concentrations indicated that the formation of higher ozone in the south and southeast of the PRD region differed from that in the Conghua area, a remote area in the north of the PRD. We recommend that the priorities for the control of VOC sources include motorcycles, gasoline vehicles, and solvent use because of their larger OFP contributions. PMID- 20028056 TI - Photochemical fate of sulfadimethoxine in aquaculture waters. AB - Sulfadimethoxine (SDM) is an antibiotic often used in combination with ormetoprim to prevent the spread of disease in freshwater aquaculture. It is known to undergo photochemical degradation in natural sunlit surface waters, but the role of dissolved organic matter (DOM) in this process is poorly understood. Our results show that water from a eutrophic catfish pond at the Mississippi State University Delta Research and Extension Center facility in Stoneville, MS facilitates the rapid phototransformation of SDM. In contrast, water from a nearby stream (Deer Creek) whose DOM is derived from allochthonous precursors does not enhance SDM photodegradation. We attribute these disparate results to DOM composition, whereby dissolved organic matter originating from highly eutrophic water bodies is a better SDM photosensitizer. Experiments conducted concurrently using respective autochthonous (Pony Lake, Antarctica) and allochthonous (Suwannee River) derived fulvic acids corroborate these findings. Scavenging experiments and experiments conducted anoxically show that the main indirect photodegradation pathway occurs by triplet excited-state DOM oxidation. Finally, transformation products assayed by mass spectrometry reveal the same major SDM photoproducts in the presence and absence of dissolved organic matter. PMID- 20028057 TI - Uranium uptake by hectorite and montmorillonite: a solution chemistry and polarized EXAFS study. AB - The mechanism of U(VI) retention on montmorillonite and hectorite at high ionic strength (0.5 M NaCl) was investigated by solution chemistry and, at near-neutral pH, polarized EXAFS spectroscopy. Uranium(VI) sorption increases from pH 3 to 7 on the two clays, but with a steeper edge for hectorite. Uranium(VI) is no longer retained at pH > 9, presumably owing to the formation of soluble anionic complexes. Polarized EXAFS showed that U(VI) retains its uranyl conformation on montmorillonite (U_mont) and hectorite (U_hect), with uranyl O at 1.79(2) A for U_mont and 1.82(2) A for U_hect, and split equatorial O shells at 2.29(2) and 2.47(2) A (U_mont), or 2.35(2) and 2.53(2) A (U_hect). An additional atomic shell of approximately 0.5 Al/Si at 3.3 A is detected for U_mont, but neither the oxygen nor the cationic shell exhibit clear angular dependence. These results indicate the formation of mononuclear complexes at the edges of montmorillonite platelets, with the orientation of the uranyl axis equal to the magic angle, as constrained by the edges' structural properties. In contrast to U_mont, the U-O signal varies with the polarization angle in U_hect, and the cationic Mg/Si contribution at 3.2 A is weak. The structure of this surface complex is not completely elucidated; it may correspond either to sorption on silanol sites, or to coprecipitation. These results lay out the fundamental molecular-scale basis to understand U retention by neoformed clay layers of nuclear glasses. PMID- 20028058 TI - Improved automated analysis of radon (222Rn) and thoron (220Rn) in natural waters. AB - Natural radon ((222)Rn) and thoron ((220)Rn) can be used as tracers of various chemical and physical processes in the environment. We present here results from an extended series of laboratory experiments intended to improve the automated analysis of (222)Rn and (220)Rn in water using a modified RAD AQUA (Durridge Inc.) system. Previous experience with similar equipment showed that it takes about 30-40 min for the system to equilibrate to radon-in-water concentration increases and even longer for the response to return to baseline after a sharp spike. While the original water/gas exchanger setup was built only for radon-in water measurement, our goal here is to provide an automated system capable of high resolution and good sensitivity for both radon- and thoron-in-water detections. We found that faster water flow rates substantially improved the response for both isotopes while thoron is detected most efficiently at airflow rates of 3 L/min. Our results show that the optimum conditions for fastest response and sensitivity for both isotopes are at water flow rates up to 17 L/min and an airflow rate of 3 L/min through the detector. Applications for such measurements include prospecting for naturally occurring radioactive material (NORM) in pipelines and locating points of groundwater/surface water interaction. PMID- 20028059 TI - Measuring the photochemical production of carbon dioxide from marine dissolved organic matter by pool isotope exchange. AB - CO(2) is the major known product of solar photolysis of marine dissolved organic matter (DOM). Measuring the rate of this globally significant process is hindered by low rates per unit volume, high background CO(2) in seawater, and ubiquitous contamination. Current methods utilize CO(2)-free seawater matrices, possibly introducing artifacts. Alternatively, pool isotope exchange (PIE) replaces most of the sample's DI(12)C with DI(13)C at natural pH and temperature, so that (12)CO(2) from DOM photooxidation elevates (12)CO(2)/(13)CO(2) ratios in irradiated samples compared to dark controls. (12)CO(2)/(13)CO(2) ratios are then measured using a modified GC-IRMS. The minimum detectable concentration change (three standard deviations) is 300 nmol DI(12)C/kg. Methods for minimizing contamination while exchanging, transferring, sealing, and irradiating samples, and for recovering and purifying CO(2) are presented. Results from PIE agree within uncertainties with those from CO(2)-free coastal seawater, suggesting that both methods apply to river-dominated coastal waters. However, photooxidation in the open ocean, which likely dominates the global flux despite lower rates per unit volume, involves DOM that differs from coastal DOM, so that coastal agreement cannot validate open-ocean studies. Major advantages of PIE are use of nearly unperturbed seawater matrices, potential to incubate samples in situ to obtain depth-integrated rates directly, and potential to use larger samples to measure open-ocean waters. PMID- 20028060 TI - Bromine chloride as an oxidant to improve elemental mercury removal from coal fired flue gas. AB - The equilibria and kinetics of the reaction between bromine (Br(2)) and chlorine (Cl(2)) to form bromine chloride (BrCl) at various temperatures were determined. BrCl was employed to oxidize elemental mercury (Hg(0)) under simulated flue gas conditions. The removal of Hg(0) from the gas phase by a homogeneous gas-phase oxidation reaction and the heterogeneous reactions involving flyash were investigated. The second-order gas phase rate constant was determined to be 2.3(+/-0.2) x 10(-17) cm(3).molecules(-1).s(-1) at 373K. The reaction of Hg(0)/BrCl was significantly accelerated in the presence of flyash, and the estimated Hg(0) removal efficiency in the presence of 0.6 ppmv BrCl and 20 g/m(3) flyash was up to 90%. Unexpectedly, the major product was found to be HgCl(2), rather than HgBr(2), indicating that bromine in part acted as the accelerant in Hg(0) oxidation in BrCl/Br(2)/Cl(2) system by facilitating the formation of intermediates. As a result, bromine consumption is much less than if only bromine gas is utilized alone. These results were helpful not only for understanding the mechanism of Hg(0) removal in coal-fired flue gas but also in any atmosphere in which bromine and chlorine species coexist. PMID- 20028061 TI - Multifunctional colloidal particles for in situ remediation of chlorinated hydrocarbons. AB - Effective in situ injection technology for the remediation of dense nonaqueous phase liquids (DNAPLs) such as trichloroethylene (TCE) requires the use of decontamination agents that effectively migrate through the soil media and react efficiently with dissolved TCE and bulk TCE. We describe the use of a novel decontamination system containing highly uniform carbon microspheres in the optimal size range for transport through the soil. The microspheres are enveloped in a polyelectrolyte (carboxymethyl cellulose, CMC) to which a bimetallic nanoparticle system of zero-valent iron and Pd is attached. The carbon serves as a strong adsorbent to TCE, while the bimetallic nanoparticle system provides the reactive component. The polyelectrolyte serves to stabilize the carbon microspheres in aqueous solution. The overall system resembles a colloidal micelle with a hydrophilic shell (polyelectrolyte coating) and hard hydrophobic core (carbon). In contact with bulk TCE, there is a sharp partitioning of the system to the TCE side of the interface due to the hydrophobicity of the core. These multifunctional systems appear to satisfy criteria related to remediation and are made with potentially environmentally benign materials. PMID- 20028062 TI - Fate of silica nanoparticles in simulated primary wastewater treatment. AB - Through novel application of small-angle neutron scattering, we examined the fate of silica nanoparticles (SiO(2)NPs) during simulated primary wastewater treatment, by measuring, in real time, the colloidal behavior of SiO(2)NPs in wastewater (sewage). We examined the effects of surface functionality on SiO(2)NP fate in wastewater, by comparing both unfunctionalized (uncoated or "bare") SiO(2)NPs and SiO(2)NPs functionalized with a thin coating of a nonionic surfactant (Tween 20), which is widely used in personal care and household product formulations containing engineered oxide nanoparticles. Our results show new evidence that the surface functionality of SiO(2)NPs plays a crucial role in their flocculation and sedimentation behavior in wastewater, and thus the likely efficacy of their removal from the effluent stream during primary wastewater treatment. Uncoated SiO(2)NPs did not flocculate in wastewater over typical residence times for primary treatment. Conversely, surface-functionalized (Tween coated) SiO(2)NPs underwent rapid flocculation in wastewater. Our results show that the surface-functionalized SiO(2)NPs are likely to be removed by sedimentation to sewage sludge (typically recycled to land), whereas uncoated SiO(2)NPs will continue through the effluent stream. While nanoparticle design is driven by use purpose, this study shows new potential for exploiting surface functionalization of nanoparticles to modify their environmental pathways. PMID- 20028063 TI - Chemical oxidation of 2,6-dimethylaniline in the fenton process. AB - 2,6-Dimethylaniline degradation by Fenton process has been studied in depth for the purpose of learning more about the reactions involved in the oxidation of 2,6 dimethylaniline under various reaction conditions. The effect of reaction conditions including the initial pH value, and the dosages of ferrous ions and hydrogen peroxide on 2,6-dimethylaniline and COD removal were investigated. 2,6 Dimethylaniline removal efficiency of 70% was achieved under optimal reaction conditions of pH value of 2, dosage of 2 mM of ferrous ion, and 20 mM of hydrogen peroxide after 3 h. A series of intermediates were identified, corresponding to ring compounds and short-chain organic acids. The intermediates were 2,6 dimethylphenol, 2,6-dimethylnitrobenzene, 2,6-dimethylbenzoquinone, 3-hexanone, maleic acid, acetic acid, formic acid, and oxalic acid. An oxidation pathway of the target organic was also proposed in this study. PMID- 20028064 TI - New metabolites in dibenzofuran cometabolic degradation by a biphenyl-cultivated Pseudomonas putida strain B6-2. AB - A biphenyl (BP)-utilizing bacterium, designated B6-2, was isolated from soil and identified as Pseudomonas putida. BP-grown B6-2 cells were capable of transforming dibenzofuran (DBF) via a lateral dioxygenation and meta-cleavage pathway. The ring cleavage product 2-hydroxy-4-(3'-oxo-3'H-benzofuran-2' yliden)but-2-enoic acid (HOBB) was detected as a major metabolite. B6-2 growing cells could also cometabolically degrade DBF using BP as a primary substrate. A recombinant Escherichia coli strain DH10B (pUC118bphABC) expressing BP dioxygenase, BP-dihydrodiol dehydrogenase, and dihydroxybiphenyl dioxygenase was shown to be capable of transforming DBF to HOBB. Using purified HOBB that was produced by the recombinant as the substrate for B6-2, we newly identified a series of benzofuran derivatives as metabolites. The structures of these metabolites indicate that an unreported HOBB degradation pathway is employed by strain B6-2. In this pathway, HOBB is proposed to be transformed to 2-oxo-4-(3' oxobenzofuran-2'-yl)butanoic acid and 2-hydroxy-4-(3'-oxobenzofuran-2' yl)butanoic acid (D4) through two sequential double-bond hydrogenation steps. D4 is suggested to undergo reactions including decarboxylation and oxidation to produce 3-(3'-oxobenzofuran-2'-yl)propanoic acid (D6). 3-Hydroxy-3-(3' oxobenzofuran-2'-yl)propanoic acid (D7) and 2-(3'-oxobenzofuran-2'-yl)acetic acid (D8) would represent metabolites involved in the processes of beta- and alpha oxidation of D6, respectively. D7 and D8 are suggested to be transformed to their respective products 3-hydroxy-2,3-dihydrobenzofuran-2-carboxylic acid (D10) and 2 (3'-hydroxy-2',3'-dihydrobenzofuran-2'-yl)acetic acid. D10 is proposed to be transformed to salicylic acid (D14) via 2,3-dihydro-2,3-dihydroxybenzofuran, 2 oxo-2-(2'-hydroxyphenyl)acetic acid and 2-hydroxy-2-(2'-hydroxyphenyl)acetic acid. Further experimental results revealed that B6-2 was capable of growing with D14 as the sole carbon source. Because benzofuran derivatives may have biological, pharmacological, and toxic properties, the elucidation of this new pathway should be significant from both biotechnological and environmental views. PMID- 20028065 TI - Comparing the environmental footprints of home-care and personal-hygiene products: the relevance of different life-cycle phases. AB - An in-depth life-cycle assessment of nine home-care and personal-hygiene products was conducted to determine the ecological relevance of different life-cycle phases and compare the environmental profiles of products serving equal applications. Using detailed data from industry and consumer-behavior studies a broad range of environmental impacts were analyzed to identify the main drivers in each life-cycle stage and potentials for improving the environmental footprints. Although chemical production significantly adds to environmental burdens, substantial impacts are caused in the consumer-use phase. As such, this research provides recommendations for product development, supply chain management, product policies, and consumer use. To reduce environmental burdens products should, for instance, be produced in concentrated form, while consumers should apply correct product dosages and low water temperatures during product application. PMID- 20028066 TI - Nutritional status and gastrointestinal microbes affect arsenic bioaccessibility from soils and mine tailings in the simulator of the human intestinal microbial ecosystem. AB - In vitro gastrointestinal models, used to measure the metal(loid) bioaccessibility for site specific risk assessment, are typically operated under fasted conditions. We evaluated the hypothesis that fed conditions increase arsenic bioaccessibility on three reference soils (NIST 2711, NIST 2709, and BGS 102) and the bulk and <38 mum size fractions of a mine tailing. The three nutritional states included a fed state with a carbohydrate mixture, a second fed state with homogenized crowberries (Empetrum nigrum), and a fasted state. The carbohydrate mixture increased arsenic bioaccessibility from four of five samples in the simulator of the human intestinal microbial ecosystem (SHIME) stomach but only three of five samples in the SHIME small intestine and colon. In contrast, crowberries increased arsenic bioaccessibility from four of five samples in the SHIME small intestine but had variable affects in the SHIME stomach and colon. The effect of nutritional status on arsenic bioaccessibility was potentially mediated via ligand-promoted dissolution in the SHIME stomach and small intestine. The displacement of arsenic with phosphate was potentially present in the SHIME small intestine but not the SHIME stomach. Microbial activity increased arsenic bioaccessibility relative to sterile conditions from four of five samples under fasted conditions and three of the five samples under fed conditions, which may suggest that in vitro gastrointestinal (GI) models operated under fed conditions and with microbes provide a more conservative estimate of in vitro bioaccessibility. However, for some samples, the arsenic bioaccessibility in the SHIME colon (with microbial activity) was equivalent to values observed in a separate physiologically based extraction test under small intestinal conditions (without microbial activity). These results suggest that the incorporation of microbial activity into in vitro GI models does not necessarily make estimates of arsenic bioaccessibility more protective than those generated using in vitro models that do not include microbial activity. PMID- 20028067 TI - Rapid changes in small fish mercury concentrations in estuarine wetlands: implications for wildlife risk and monitoring programs. AB - Small fish are commonly used to assess mercury (Hg) risk to wildlife and monitor Hg in wetlands. However, limited research has evaluated short-term Hg variability in small fish, which can have important implications for monitoring programs and risk assessment. We conducted a time-series study of Hg concentrations in two small fish species representing benthic (longjaw mudsuckers [Gillichthys mirabilis]) and pelagic (threespine sticklebacks [Gasterosteus aculeatus]) food webs within three wetland habitats in San Francisco Bay Estuary. We simultaneously monitored prey deliveries, nest initiation, and chick hatching dates of breeding Forster's terns (Sterna forsteri), the most abundant nesting piscivore in the region. Mudsuckers and sticklebacks were the predominant prey fish, comprising 36% and 25% of tern diet, and Hg concentrations averaged (geometric mean +/- SE, microg/g dw) 0.44 +/- 0.01 and 0.68 +/- 0.03, respectively. Fish Hg concentrations varied substantially over time following a quadratic form in both species, increasing 40% between March and May then decreasing 40% between May and July. Importantly, Forster's terns initiated 68% of nests and 31% of chicks hatched during the period of peak Hg concentrations in prey fish. These results illustrate the importance of short-term temporal variation in small fish Hg concentrations for both Hg monitoring programs and assessing wildlife risk. PMID- 20028068 TI - Influence of prey type on nickel and thallium assimilation, subcellular distribution and effects in juvenile fathead minnows (Pimephales promelas). AB - Because fish take up metals from prey, it is important to measure factors controlling metal transfer between these trophic levels so as to explain metal bioaccumulation and effects in fish. To achieve this, we exposed two types of invertebrates, an oligochaete (Tubifex tubifex) and a crustacean (Daphnia magna), to environmentally relevant concentrations of two important contaminants, nickel (Ni) and thallium (Tl), and fed these prey to juvenile fathead minnows (Pimephales promelas). We then measured the assimilation efficiency (AE), subcellular distribution and effects of these metals in fish. Fish assimilated dietary Tl more efficiently from D. magna than from T. tubifex, and more efficiently than Ni, regardless of prey type. However, the proportion of metal bound to prey subcellular fractions that are likely to be trophically available (TAM) had no significant influence on the efficiency with which fish assimilated Ni or Tl. In fish, the majority of their Ni and Tl was bound to subcellular fractions that are purportedly detoxified, and prey type had a significant influence on the proportion of detoxified Ni and Tl in fish. We measured higher activities of cytochrome C oxidase and glutathione S-transferase in fish fed D. magna compared to fish fed T. tubifex, regardless of the presence or absence of Ni or Tl in prey. However, we measured decreased activities of glutathione S transferase and nucleoside diphosphate kinase in fish fed Tl-contaminated D. magna compared to fish from the three other treatment levels. PMID- 20028069 TI - Influence of substrate on electron transfer mechanisms in chambered benthic microbial fuel cells. AB - This research investigated whether the addition of an exogenous electron donor would affect power production in laboratory-scale benthic microbial fuel cells (BMFC) by differentially influencing microbially mediated electron transfer processes. Six BMFCs were operated for over one year in a temperature-controlled laboratory. Three BMFCs relied on endogenous electron donors, and three were supplemented with lactate. The supplemented BMFCs generated more cumulative charge, but did not generate higher average current between periods of lactate enrichment. Coulombic efficiencies during the lactate treatments ranged from 25 to 65% suggesting that lactate utilization was variably coupled to power production. Cumulative electron flux resulting from lactate additions and chemical changes within the anode chamber, as well as a difference in the anode hosted microbial communities indicated that lactate supplementation promoted sulfate reduction. After the addition of molybdate to suppress sulfate reduction and sulfur disproportionation, all BMFCs continued to produce current, but no longer responded to lactate additions. Chemical data support a two-step cycle in which endogenous organic carbon and/or supplemented lactate fuel sulfate reduction resulting in sulfide and simple organic molecules (such as acetate) that can act as the electron donors for the BMFC. PMID- 20028070 TI - Soil carbon sequestration or biofuel production: new land-use opportunities for mitigating climate over abandoned Soviet farmlands. AB - Although the CO(2) mitigation potential of biofuels has been studied by extrapolation of small-scale studies, few estimates exist of the net regional scale carbon balance implications of biofuel cultivations programs, either growing conventional biofuel crops or applying new advanced technologies. Here we used a spatially distributed process-driven model over the 20 Mha of recently abandoned agricultural lands of the Former Soviet Union to quantify the GHG mitigation by biofuel production from Low Input/High Diversity (LIHD) grass legume prairies and to compare this GHG mitigation with the one of soil C sequestration as it currently occurs. LIHD has recently received a lot of attention as an emerging opportunity to produce biofuels over marginal lands leading to a good energy efficiency with minimal adverse consequences on food security and ecosystem services. We found that, depending on the time horizon over which one seeks to maximize the GHG benefit, the optimal time for implementing biofuel production shifts from "never" (short-term horizon) to "as soon as possible" (longer-term horizon). These results highlight the importance of reaching agreement a priori on the target time interval during which biofuels are expected to play a role within the global energy system, to avoid deploying biofuel technology over a time interval for which it has a detrimental impact on the GHG mitigation objective. The window of opportunity for growing LIHD also stresses the need to reduce uncertainties in soil C inputs, turnover, and soil organic matter stability under current and future climate and management practices. PMID- 20028071 TI - Bioelectrocatalysis of Acetobacter aceti and Gluconobacter roseus for current generation. AB - Acetobacter aceti and Gluconobacter roseus, which are known to be responsible for the spoilage of wine, are used for current generation in batch-type microbial biofuel cells and it has been shown for the first time that these two microorganisms do not require mediators for the transfer of electrons to the anode. Three biofuel cells were constructed with two cells containing the pure cultures of each of the microorganisms as the biocatalyst (A-MFC, G-MFC) and the third cell was constructed with the mixed culture of these two microorganisms as the biocatalyst (AG-MFC). The performance of the biofuel cells was evaluated in terms of open circuit voltage (OCV), fuel consumption rate, internal resistance, power output, and coulombic efficiency. The mixed culture cell (AG-MFC) exhibits a better overall performance compared to the other cells. PMID- 20028072 TI - Nitrobenzene removal in bioelectrochemical systems. AB - Nitrobenzene occurs as a pollutant in wastewaters originating from numerous industrial and agricultural activities. It needs to be removed prior to discharge to sewage treatment works because of its high toxicity and persistence. In this study, we investigated the use of a bioelectrochemical system (BES) to remove nitrobenzene at a cathode coupled to microbial oxidation of acetate at an anode. Effective removal of nitrobenzene at rates up to 1.29 +/- 0.04 mol m(-3) TCC d( 1) (total cathodic compartment, TCC) was achieved with concomitant energy recovery. Correspondingly, the formation rate for the reduction product aniline was 1.14 +/- 0.03 mol m(-3) TCC d(-1). Nitrobenzene removal and aniline formation rates were significantly enhanced when the BES was supplied with power, reaching 8.57 +/- 0.03 and 6.68 +/- 0.03 mol m(-3) TCC d(-1), respectively, at an energy consumption of 17.06 +/- 0.16 W m(-3) TCC (current density at 59.5 A m(-3) TCC). Compared to those of conventional anaerobic biological methods for nitrobenzene removal, the required dosage of organic cosubstrate was significantly reduced in this system. Although aniline was always identified as the major product of nitrobenzene reduction at the cathode of BES in this study, the Coulombic efficiencies of nitrobenzene removal and aniline formation were dependent on the current density of the BES. PMID- 20028074 TI - Comment on "effect of groundwater iron and phosphate on the efficacy of arsenic removal by iron-amended biosand filters". PMID- 20028076 TI - Comment on "occurrence and potential significance of perfluorooctanoic acid (PFOA) detected in New Jersey public drinking water systems". PMID- 20028078 TI - Mass spectrometry-based identification of Y745 of Vav1 as a tyrosine residue crucial in maturation of acute promyelocytic leukemia-derived cells. AB - Vav1, whose physiological expression is restricted to hematopoietic system, is one of the signaling proteins up-regulated by all-trans retinoic acid (ATRA) in acute promyelocytic leukemia (APL)-derived precursors, in which it promotes the overcoming of the differentiation blockade. High levels of tyrosine phosphorylated Vav1 accumulate in differentiating APL-derived cells, suggesting that one or more Vav1 tyrosine residues are involved in neutrophil differentiation of tumoral promyelocytes. Here, we have found that phosphorylation of Vav1 Y174, that is known to regulate Vav1 activity in mature neutrophils, is up-regulated by ATRA in NB4 cells. Nevertheless, this tyrosine residue does not seem crucial for the agonist-induced phenotypical differentiation of APL-derived cells. Mass spectrometry analysis performed on Vav1 from differentiating NB4 cells allowed to identify the highly conserved Y745 residue as a phosphorylated tyrosine that plays crucial roles in the completion of the maturation program of this cell line. In fact, the overexpression of a mutated form of Vav1, in which Y745 was replaced with a phenylalanine, significantly reduced the ATRA-induced CD11b expression and essentially abrogated the differentiation-related acquisition of the migratory capability. Even though the intracellular signaling involving Vav1 phosphorylated in Y745 is unknown, the identification of a tyrosine residue essential for differentiation of tumoral precursors may constitute the basis to identify new specific targets for differentiation therapy of APL. PMID- 20028079 TI - Comprehensive identification of staurosporine-binding kinases in the hepatocyte cell line HepG2 using Capture Compound Mass Spectrometry (CCMS). AB - The central role of kinases in cell signaling has set them in the focus of biomedical research. In functional proteomics analyses, large- scale profiling of kinases has become feasible through the use of affinity pulldown beads that carry immobilized kinase inhibitors. As an alternative approach to solid phase beads, Capture Compound Mass Spectrometry (CCMS) enables the functional isolation of protein-classes on the basis of small molecule-protein interactions in solution. Capture Compounds are trifunctional probes: a selectivity function interacts with the native target proteins in equilibrium, upon irradiation a photoactivatable reactivity function forms an irreversible covalent bond to the target proteins, and a sorting function allows the captured proteins to be isolated from a complex protein mixture. We report the design and application of a novel, fully water soluble Capture Compound that carries the broadband kinase inhibitor staurosporine as selectivity function. We used this Capture Compound to profile the kinome of the human liver-derived cell line HepG2 and identified one hundred kinases. HepG2 cells are a widely used model system for hepatocarcinoma, hepatitis, and for investigation of drug toxicity effects. CCMS experiments in membrane fractions of human placenta are given as example for the applicability to human tissue. PMID- 20028081 TI - Micropeptins from an Israeli fishpond water bloom of the cyanobacterium Microcystis sp. AB - Seven new natural products, micropeptin MZ845 (1), micropeptin MZ859 (2), micropeptin MZ939A (3), micropeptin MZ925 (4), micropeptin MZ939B (5), micropeptin MZ1019 (6), and micropeptin MZ771 (7), as well as two known micropeptins, cyanopeptolin S (8) and cyanopeptolin SS (9), were isolated from the hydrophilic extract of the cyanobacterium Microcystis sp. that was collected from a fishpond in Kibbutz Ma'ayan Tzvi, Israel, in July 2006. The structures of the pure natural products were elucidated using spectroscopic methods, including UV, 1D and 2D NMR, and MS techniques. The absolute configuration of the chiral centers of the compounds was determined using Marfey's method for HPLC. The inhibitory activity of the compounds was determined for the serine proteases: trypsin, chymotrypsin, thrombin, and elastase. These micropeptins inhibited trypsin with IC(50)'s that varied between 0.6 and 24.2 microM. The SAR of these micropeptins is discussed. PMID- 20028080 TI - Expression of Vibrio harveyi acyl-ACP synthetase allows efficient entry of exogenous fatty acids into the Escherichia coli fatty acid and lipid A synthetic pathways. AB - Although the Escherichia coli fatty acid synthesis (FAS) pathway is the best studied type II fatty acid synthesis system, a major experimental limitation has been the inability to feed intermediates into the pathway in vivo because exogenously supplied free fatty acids are not efficiently converted to the acyl acyl carrier protein (ACP) thioesters required by the pathway. We report that expression of Vibrio harveyi acyl-ACP synthetase (AasS), a soluble cytosolic enzyme that ligates free fatty acids to ACP to form acyl-ACPs, allows exogenous fatty acids to enter the E. coli fatty acid synthesis pathway. The free fatty acids are incorporated intact and can be elongated or directly incorporated into complex lipids by acyltransferases specific for acyl-ACPs. Moreover, expression of AasS strains and supplementation with the appropriate fatty acid restored growth to E. coli mutant strains that lack essential fatty acid synthesis enzymes. Thus, this strategy provides a new tool for circumventing the loss of enzymes essential for FAS function. PMID- 20028082 TI - 1,3-Diazepanes of natural product-like complexity from cyanamide-induced rearrangement of epoxy-delta-lactams. AB - A synthetic procedure toward 1,3-diazepane scaffolds of natural product-like complexity was developed for the construction of RNA-directed ligand libraries. A molecular building block was designed that combines the characteristics of RNA binding natural products, including a high density of hydrogen bond donors and acceptors around a rigid, nonplanar scaffold with straightforward total-synthetic accessibility that permits extensive control over the chemical space. The synthesis of the 1,3-diazepane scaffold was achieved via an unprecedented cyanamide-induced rearrangement of epoxy-delta-lactams. PMID- 20028084 TI - Kinetic mechanism of DNA unwinding by the BLM helicase core and molecular basis for its low processivity. AB - Bloom's syndrome (BS) is a rare human autosomal recessive disorder characterized by a strong predisposition to a wide range of cancers commonly affecting the general population. Understanding the functioning mechanism of the BLM protein may provide the opportunity to develop new effective therapy strategies. In this work, we studied the DNA unwinding kinetic mechanism of the helicase core of the BLM protein using various stopped-flow assays. We show that the helicase core of BLM unwinds duplex DNA as monomers even under conditions strongly favoring oligomerization. An unwinding rate of approximately 20 steps per second and a step size of 1 bp have been determined. We have observed that the helicase has a very low processivity. From dissociation and inhibition experiments, we have found that during its ATP hydrolysis cycle in DNA unwinding the helicase tends to dissociate from the DNA substrate in the ADP state. The experimental results imply that the BLM helicase core may unwind duplex DNA in an inchworm manner. PMID- 20028083 TI - Photo-cross-linking of XPC-Rad23B to cisplatin-damaged DNA reveals contacts with both strands of the DNA duplex and spans the DNA adduct. AB - Nucleotide excision repair (NER) is the main pathway used for the repair of bulky DNA adducts such as those caused by UV light exposure and the chemotherapeutic drug cisplatin. The xeroderma pigmentosum group C (XPC)-Rad23B complex is involved in the recognition of these bulky DNA adducts and initiates the global genomic nucleotide excision repair pathway (GG-NER). Photo-cross-linking experiments revealed that the human XPC-Rad23B complex makes direct contact with both the cisplatin-damaged DNA strand and the complementary undamaged strand of a duplex DNA substrate. Coupling photo-cross-linking with denaturation and immunoprecipitation of protein-DNA complexes, we identified the XPC subunit in complex with damaged DNA. While the interaction of the XPC subunit with DNA was direct, studies revealed that although Rad23B was found in complex with DNA, the Rad23B-DNA interaction was largely indirect via its interaction with XPC. Using site specific cross-linking, we determined that the XPC-Rad23B complex is preferentially cross-linked to the damaged DNA when the photoreactive FAP-dCMP (exo-N-{2-[N-(4-azido-2,5-difluoro-3-chloropyridin-6-yl)-3 aminopropionyl]aminoethyl}-2'-deoxycytidine 5'-monophosphate) analogue is located to the 5' side of the cisplatin-DNA adduct. When the FAP-dCMP analogue is located to the 3' side of the adduct, no difference in binding was detected between undamaged and damaged DNA. Collectively, these data suggest a model in which XPC DNA interactions drive the damage recognition process contacting both the damaged and undamaged DNA strand. Preferential cross-linking 5' of the cisplatin-damaged site suggests that the XPC-Rad23B complex displays orientation specific binding to eventually impart directionality to the downstream binding and incision events relative to the site of DNA damage. PMID- 20028085 TI - Binding affinity and kinetic analysis of targeted small molecule-modified nanoparticles. AB - Nanoparticles bearing surface-conjugated targeting ligands are increasingly being explored for a variety of biomedical applications. The multivalent conjugation of targeting ligands on the surface of nanoparticles is presumed to enhance binding to the desired target. However, given the complexities inherent in the interactions of nanoparticle surfaces with proteins, and the structural diversity of nanoparticle scaffolds and targeting ligands, our understanding of how conjugation of targeting ligands affects nanoparticle binding remains incomplete. Here, we use surface plasmon resonance (SPR) to directly and quantitatively study the affinity and binding kinetics of nanoparticles that display small molecules conjugated to their surface. We studied the interaction between a single protein target and a structurally related series of targeting ligands whose intrinsic affinity varies over a 4500-fold range and performed SPR at protein densities that reflect endogenous receptor densities. We report that even weak small molecule targeting ligands can significantly enhance target-specific avidity (by up to 4 orders of magnitude) through multivalent interactions and also observe a much broader range of kinetic effects than has been previously reported. Quantitative measurement of how the affinity and kinetics of nanoparticle binding vary as a function of different surface conjugations is a rapid, generalizable approach to nanoparticle characterization that can inform the design of nanoparticles for biomedical applications. PMID- 20028086 TI - A metabolomics study of cultivated potato (Solanum tuberosum) groups Andigena, Phureja, Stenotomum, and tuberosum using gas chromatography-mass spectrometry. AB - Phytochemical diversity was examined by gas chromatography-mass spectrometry in tubers of genotypes belonging to groups Andigena, Phureja, Stenotomum, and Tuberosum of the potato, Solanum tuberosum. Polar extracts (mainly amino acids, organic acids, sugars, and sugar alcohols) and nonpolar extracts (mainly fatty acids, fatty alcohols, and sterols) were examined. There was a large range in levels of metabolites, including those such as asparagine, fructose, and glucose, that are important to tuber quality, offering considerable scope for selecting germplasm for breeding programmes. There were significant differences in the levels of many metabolites among the groups. The metabolite profiles of genotypes belonging to Phureja and Stenotomum were similar and different from those of Tuberosum and the majority of Andigena genotypes. There was some agreement with the phylogeny of the groups in that Stenotomum is believed to be the ancestor of Phureja and they are both distinct from Tuberosum. Andigena genotypes could be partially distinguished according to geographical origin, Bolivian genotypes being particularly distinct from those from Ecuador. Biosynthetic links between metabolites were explored by performing pairwise correlations of all metabolites. The significance of some expected and unexpected strong correlations between many amino acids (e.g., between isoleucine, lysine, valine, and other amino acids) and between several nonpolar metabolites (e.g., between many fatty acids) is discussed. For polar metabolites, correlation analysis gave essentially similar results irrespective of whether the whole data set, only Andigena genotypes, or only Phureja genotypes were used. In contrast, for the nonpolar metabolites, Andigena only and Phureja only data sets resulted in weaker and stronger correlations, respectively, compared to the whole data set, and may suggest differences in the biochemistry of the two groups, although the interpretation should be viewed with some caution. PMID- 20028087 TI - Direct electrochemistry of drug metabolizing human flavin-containing monooxygenase: electrochemical turnover of benzydamine and tamoxifen. AB - This communication reports on the first electrochemical study of the human flavin containing monooxygenase 3 (hFMO3) either absorbed or covalently linked to different electrode surfaces. Glassy carbon and gold electrodes gave reversible electrochemical signals of an active hFMO3. The midpoint potential measured for the immobilized enzyme on a glassy carbon electrode was -445 +/- 8 mV (versus Ag/AgCl). A monolayer coverage was obtained on gold functionalized with dithio bismaleimidoethane that covalently linked surface accessible cysteines of hFMO3. A structural model of the enzyme was generated to rationalize electrochemistry results. The turnover of the active enzyme was measured with two specific drugs: tamoxifen and benzydamine. For tamoxifen, 1.7 and 8.0 microM of its N-oxide product were formed by the enzyme immobilized on glassy carbon and gold electrodes, respectively. In the case of benzydamine, a K(M) of 44 +/- 5 microM was measured upon application of a -600 mV bias to the enzyme immobilized on the glassy carbon electrode that is in good agreement with the values published for microsomal hFMO3 where NADPH is the electron donor. PMID- 20028088 TI - Phi-value analysis for ultrafast folding proteins by NMR relaxation dispersion. AB - Proteins that fold rapidly, on the (sub-) microsecond time scale, offer the prospect of direct comparison between experimental data and molecular dynamics simulations. However, experimental studies for such proteins frequently are hindered because folding rates are too fast to measure using conventional stopped flow methods. To overcome this impediment, NMR spin relaxation dispersion experiments are used to quantify mutational effects on kinetics (DeltaDeltaG(o)), stability (DeltaDeltaG(o)), and phi-values (DeltaDeltaG(dagger)/DeltaDeltaG(o)) for proteins exhibiting chemical exchange line broadening that is fast on the NMR chemical shift time scale. The accuracy of phi-value analysis is enhanced because mutational effects on denatured or intermediate states can be detected through changes in line broadening. The transition and intermediate states of the villin headpiece domain, HP67, are characterized in varying solvent conditions to validate the method. PMID- 20028089 TI - Plasmon-induced photodegradation of toxic pollutants with Ag-AgI/Al2O3 under visible-light irradiation. AB - A plasmonic photocatalyst Ag-AgI supported on mesoporous alumina (Ag AgI/Al(2)O(3)) was prepared by deposition-precipitation and photoreduction methods. The catalyst showed high and stable photocatalytic activity for the degradation and mineralization of toxic persistent organic pollutants, as demonstrated with 2-chlorophenol (2-CP), 2,4-dichlorophenol (2,4-DCP), and trichlorophenol (TCP) under visible light or simulated solar light irradiation. On the basis of electron spin resonance, cyclic voltammetry analyses under a variety of experimental conditions, two electron transfer processes were verified from the excited Ag NPs to AgI and from 2-CP to the Ag NPs, and the main active species of O(2)(*-) and excited h(+) on Ag NPs were involved in the photoreaction system of Ag-AgI/Al(2)O(3). A plasmon-induced photocatalytic mechanism was proposed. Accordingly, the plasmon-induced electron transfer processes elucidated the photostability of Ag-AgI/Al(2)O(3). This finding indicates that the high photosensitivity of noble metal NPs due to surface plasmon resonance could be applied toward the development of new plasmonic visible-light-sensitive photocatalysts and photovoltaic fuel cells. PMID- 20028090 TI - Practical one-pot two-step protocol for the microwave-assisted synthesis of highly functionalized rhodanine derivatives. AB - A fast and efficient protocol for the generation of substituted 5-arylidene rhodanines in a sequential one-pot two-step process combining the "Holmberg method" and the Knoevenagel condensation under microwave-assisted conditions has been developed. The final compounds 11a-k have been obtained in high yield and purity after a simple precipitation from methanol, making this procedure facile, practical, and rapid to execute. PMID- 20028091 TI - Ecological control of fecal indicator bacteria in an urban stream. AB - Fecal indicator bacteria (FIB) have long been used as a marker of fecal pollution in surface waters subject to point source and non-point source discharges of treated or untreated human waste. In this paper, we set out to determine the source(s) of elevated FIB concentrations in Cucamonga Creek, a concrete-lined urban stream in southern California. Flow in the creek consists primarily of treated and disinfected wastewater effluent, mixed with relatively smaller but variable flow of runoff from the surrounding urban landscape. Dry and wet weather runoff contributes nearly 100% of FIB loading to Cucamonga Creek, while treated wastewater contributes significant loading of nutrients, including dissolved organic carbon (DOC), phosphorus, nitrate, and ammonium. FIB concentrations are strongly positively correlated with DOC concentration in runoff (Spearman's rho >or= 0.66, P Co(III) or Co(III) --> Co(II) processes, but this is not the case for other electron transfer reactions where significant differences are seen between the examined compounds. Significant interactions are also observed between the two porphyrin macrocycles of the laterally bridged dicobalt(II) bis-porphyrin dyad Co(P)-TA-(P)Co in its singly and doubly reduced form, but only weak interactions are seen between the two Co(PQ) units of the single bond biquinolalinyl-bridged dicobalt(II) bis-porphyrin dyad Co(PQ)-(QP)Co. PMID- 20028097 TI - Peptide--silica hybrid networks: biomimetic control of network mechanical behavior. AB - Self-assembly represents a robust and powerful paradigm for the bottom-up construction of nanostructures. Templated condensation of silica precursors on self-assembled nanoscale peptide fibrils with various surface functionalities can be used to mimic biosilicification. This template-defined approach toward biomineralization was utilized for the controlled fabrication of 3D hybrid nanostructures. The peptides MAX1 and MAX8 used herein form networks consisting of interconnected, self-assembled beta-sheet fibrils. We report a study on the structure--property relationship of self-assembled peptide hydrogels where mineralization of individual fibrils through sol--gel chemistry was achieved. The nanostructure and consequent mechanical characteristics of these hybrid networks can be modulated by changing the stoichiometric parameters of the sol--gel process. The physical characterization of the hybrid networks via electron microscopy and small-angle scattering is detailed and correlated with changes in the network mechanical behavior. The resultant high fidelity templating process suggests that the peptide substrate can be used to template the coating of other functional inorganic materials. PMID- 20028098 TI - Efficient electron transfer in functional assemblies of pyridine-modified NQDs on SWNTs. AB - Nanocrystal quantum dot (NQD)/single-walled carbon nanotube (SWNT) hybrid nanomaterials were synthesized, assembled into field effect transistors (FETs) via dielectrophoresis (DEP), and characterized optically and electronically. The pyridine moiety functioned as a short, noncovalent linker between the NQDs and SWNTs and allowed more efficient carrier transfer through the assemblies without deleteriously altering electronic structures. Photoluminescence studies of the resulting assemblies support an efficient carrier transfer process in CdSe-py SWNTs unlike that of CdSe/ZnS-py-SWNTs. The use of DEP as a means of controlling the assembly process allowed the creation of a SWNT array containing densely packed CdSe NQDs across a 2 mum gap between electrodes. Observations and characterization of the photocurrent, resistivity, gate dependence, and optical properties of these systems suggest efficient electron transfer from photoexcited NQDs to SWNTs. PMID- 20028099 TI - Uncovering individual hydrogen bonds in rotaxanes by frequency shifts. AB - We present a theoretical investigation of amide pseudorotaxane IR spectra in the harmonic approximation. In particular, we focus on the effect of axle substitution on the hydrogen bonds that are formed between axle and wheel. Two types of pseudorotaxanes are studied: one with the substituent affecting mostly the axle's carbonyl group and one with the effect influencing primarily the amide NH group. Sizeable red shifts are predicted for the carbonyl stretching frequencies, and large red shifts for the NH stretching frequencies. For the wheel amide groups involved in hydrogen bonding merely with their NH hydrogens, a small shift is observed for the carbonyl stretch mode. A clear relation is observed between the NH stretch shifts and individual hydrogen bond energies. This is confirmed by correlations of the shared electron number with the NH stretch shift showing that this quantity can be taken as an indicator for individual hydrogen bond energies. Axle substitution influences the strengths of the individual hydrogen bonds which is again reflected in the NH stretch frequency shifts. A linear relationship of Hammett's substituent parameters with the NH frequency shifts can be established. PMID- 20028100 TI - The coumarin-binding site in carbonic anhydrase accommodates structurally diverse inhibitors: the antiepileptic lacosamide as an example and lead molecule for novel classes of carbonic anhydrase inhibitors. AB - Coumarins constitute a general and totally new class of inhibitors of the zinc enzyme carbonic anhydrase (CA, EC 4.2.1.1), binding at the entrance of the active site cavity. We report here that the coumarin-binding site in CAs may interact with diverse compounds, such as the antiepileptic drug lacosamide, which inhibits mammalian CAs I-XV, with inhibition constants in range of 331 nM to 4.56 microM. Its X-ray crystal structure in adduct with CA II reveals the molecular basis for this inhibition. Lacosamide was found in the coumarin-binding site, making favorable van der Waals interactions with Thr200, Asn67, Gln92, and Phe131. No interactions with the Zn(II) ion were evidenced in the CA II-lacosamide adduct. The coumarin-binding site may thus accommodate structurally diverse compounds which possess an inhibition mechanism distinct of that of sulfonamides. This finding opens new possibilities for designing CA inhibitors/activators with various biomedical applications. PMID- 20028101 TI - Geometry dependence of surface plasmon polariton lifetimes in nanohole arrays. AB - We study surface plasmon polariton lifetimes in two-dimensional arrays of blind holes on gold surfaces. The lifetimes are determined from the line widths of the resonant dips appearing in the specular reflection spectra. We find a strong dependence of lifetimes on the resonant wavelength and the hole geometry. Through both experiments and numerical simulations, we analyze the validity of the Rayleigh approximation and also explore the range of wavelengths and geometric parameters where it fails. Finally, we show that, within the range of geometries and wavelengths considered in our experiments, the behavior of surface plasmon polariton lifetimes can be understood as resulting from the interplay between the intrinsic metal absorption and the scattering of surface waves by single isolated holes. PMID- 20028102 TI - Large-area ordered superlattices from magnetic Wustite/cobalt ferrite core/shell nanocrystals by doctor blade casting. AB - Although a large diversity of single-component and binary superlattices from colloidal nanocrystals have been demonstrated, applications of such ordered nanocrystal assemblies are still hampered due to a lack of control over the self assembly processes over large areas. A reel-to-reel compatible large-area coating technique for solutions is given by doctor blade casting, which is applied here to deposit colloidal nanocrystals onto various substrates. The self-assembly process is demonstrated for magnetic nanocrystals, having a high potential for applications in magnetic memory devices. Shape-controlled (spherical and cubic) and monodisperse nanocrystals with a Wustite core and a cobalt ferrite shell are used in particular. Doctor blade casting of these colloidal nanocrystals results in films exhibiting hexagonally closely packed arrangements, which are formed by a top-down growth, as is evidenced by cross sectional transmission electron microscopy. The ordering in the topmost layer extends over large areas, although some defects and irregularities are found. The degree and quality of self assembly is quantified by analyzing plan view images of the assemblies by means of the decay of their autocorrelation function. This analysis reveals that the degree of ordering obtained by doctor blade casting outperforms those provided by alternative deposition techniques such as inkjet printing or drop casting. The results for the coherent lengths deduced from the autocorrelation analysis are shown to be consistent with those from grazing-incidence small-angle X-ray scattering, giving coherence length on the order of 1000 nm. PMID- 20028103 TI - Ultrafast growth of highly branched palladium nanostructures for catalysis. AB - Palladium is widely used as a catalyst in pharmaceutical and chemical syntheses as well as in the reduction of harmful exhaust emissions. Therefore, the development of high performance palladium catalysts is an area of major concern. In this paper, we present the synthesis of highly branched palladium nanostructures in a simple solution phase reaction at room temperature. By varying the nature of the organic stabilizer system we demonstrate control over the reaction kinetics and hence the shape of the nanostructures. Investigations into the structural evolution of the nanostructures show that they form from multiply twinned face centered cubic (fcc) nanoparticle nuclei. Reaction kinetics then determine the resulting shape where ultrafast growth is shown to lead to the highly branched nanostructures. These results will contribute greatly to the understanding of complex nanoparticle growth from all fcc metals. The nanostructures then show excellent catalytic activity for the hydrogenation of nitrobenzene to aniline. PMID- 20028104 TI - Lewis-base-stabilized dichlorosilylene: a two-electron sigma-donor ligand. AB - The first structurally described cobalt(I) Lewis-base-stabilized silylene complex [Co(CO)(3){SiCl(2)(IPr)}(2)](+)[CoCl(3)(THF)](-) [1; IPr = 1,3-bis(2,6 diisopropylphenyl)imidazol-2-ylidene] was prepared by applying the two-electron sigma-donor ligand SiCl(2)(IPr) through coordination with Co(2)(CO)(8). The bonding situation between ligand SiCl(2)(IPr) and the cobalt(I) metal center in [Co(CO)(3){SiCl(2)(IPr)}(2)](+) of 1 was investigated by (1)H NMR and IR spectroscopy, single-crystal X-ray structural analysis, and density functional theoretical calculations. PMID- 20028105 TI - Synthesis of graphene on silicon dioxide by a solid carbon source. AB - We report on a method for the fabrication of graphene on a silicon dioxide substrate by solid-state dissolution of an overlying stack of a silicon carbide and a nickel thin film. The carbon dissolves in the nickel by rapid thermal annealing. Upon cooling, the carbon segregates to the nickel surface forming a graphene layer over the entire nickel surface. By wet etching of the nickel layer, the graphene layer was allowed to settle on the original substrate. Scanning tunneling microscopy (STM) as well as Raman spectroscopy has been performed for characterization of the layers. Further insight into the morphology of the layers has been gained by Raman mapping indicating micrometer-size graphene grains. Devices for electrical measurement have been manufactured exhibiting a modulation of the transfer current by backgate electric fields. The presented approach allows for mass fabrication of polycrystalline graphene without transfer steps while using only CMOS compatible process steps. PMID- 20028106 TI - Steroidal saponins from Chlorophytum orchidastrum. AB - Six new spirostane-type saponins (1-6), named orchidastrosides A-F, and chloromaloside D were isolated from an ethanol extract of the roots of Chlorophytum orchidastrum. The saponins have neotigogenin or neogitogenin as the aglycon and oligosaccharidic chains possessing seven to nine sugar units. Their structures were elucidated mainly by 2D NMR spectroscopic analyses (COSY, TOCSY, NOESY, HSQC, and HMBC) and FABMS and HRESIMS. Compounds 1-6 were tested for cytotoxicity against two human colon cancer cell lines, HCT 116 and HT-29. PMID- 20028107 TI - Uniting anion relay chemistry with Pd-mediated cross coupling: design, synthesis and evaluation of bifunctional aryl and vinyl silane linchpins. AB - Union of type II Anion Relay Chemistry (ARC) with Pd-induced Cross Coupling Reactions (CCR) has been achieved, in conjunction with the design, synthesis, and evaluation of a new class of bifunctional linchpins, comprising a series of vinyl silanes bearing beta- or gamma-electrophilic sites. The synthetic tactic permits both alkylation and Pd-mediated CCR of the anions derived via 1,4-silyl C(sp(2))- >O Brook Rearrangements. PMID- 20028108 TI - Cytotoxic 3,4-seco-cycloartane triterpenes from the exudate of Gardenia tubifera. AB - Four new 3,4-seco-cycloartanes, gardenoins A-D (1-4), together with the known compound secaubryenol (5), were isolated from the exudate of Gardenia tubifera. The structures of 1-4 were elucidated on the basis of spectroscopic analysis. The cytotoxic activity of compounds 1-4 was evaluated against five human tumor cell lines. PMID- 20028109 TI - Family of V(III)-tristhiolato complexes relevant to functional models of vanadium nitrogenase: synthesis and electronic structure investigations by means of high frequency and -field electron paramagnetic resonance coupled to quantum chemical computations. AB - A series of V(III) complexes of varying coordination number (5, 6, and 7) all containing the PS3 ligand (PS3 = trianion of tris(2-thiophenyl)phosphine and its derivatives with other phenyl substituents) has been prepared and structurally characterized. The complexes have general formula [V(PS3)L(n)](0,-), where n = 1 (from L = Cl(-), 1-Me-Im, N(3)(-)), 2 (from L = 2,2'-bpy; counting each N of the bidentate ligand), and 3 (from L = 1-Me-Im, N(2)H(4)). The complexes have also been investigated by direct current (DC) magnetic susceptibility and high frequency and -field electron paramagnetic resonance (HFEPR). HFEPR, supported by magnetometry, has provided accurate spin Hamiltonian parameters that describe the S = 1 spin ground state of the complexes. Of particular interest are the zero field splitting (zfs) parameters which, together with structural data, are the empirical starting point for detailed computational studies. The computational methods included density functional theory (DFT), which was only marginally successful, and more advanced ab initio methods (CASSCF and SORCI). The zfs in these complexes is relatively small in magnitude (|D| approximately 1 cm(-1)) and is the result of multiple, often counteracting, spin-orbit coupling (SOC) and spin-spin coupling (SSC) contributions. The specific origin of each of these contributions is described in detail. The results indicate the level of electronic structure calculation possible for transition metal complexes even with multiple unpaired electrons and highly covalent, heavier atom donor ligands. PMID- 20028110 TI - Template synthesis of iron(II) complexes containing tridentate P-N-S, P-N-P, P-N N, and tetradentate P-N-N-P ligands. AB - A series of mer-tridentate iron(II) complexes bearing P-N-S (3), P-N-P (4), and P N-N (5) ligands have been prepared via the metal template effect in one pot involving air-stable phosphonium dimers [cyclo-(-PPh(2)CH(2)C(OH)H-)(2)](Br)(2) (1) and [cyclo-(-PCy(2)CH(2)C(OH)H-)(2)](Br)(2) (2), KOtBu, [Fe(H(2)O)(6)][BF(4)](2) and 2-aminothiolphenol (for 3), 2 (diphenylphosphino)ethylamine (for 4), and 2-(aminomethyl)pyridine (for 5). The new phosphonium dimer 2 was prepared via an S(N)2 reaction of PCy(2)H with BrCH(2)CH(OEt)(2). The complexes Fe{PR(2)CH(2)CH=N(2-C(6)H(4))S}(2)FeBr(2) (3a, R = Ph; 3b, R = Cy) are paramagnetic, and X-ray diffraction studies revealed that they are bimetallic, in which the S atoms of the bis-tridentate (PNS)(2)Fe unit bridge to a FeBr(2) fragment. Complexes [Fe(PR(2)CH(2)CH=NC(2)H(4)PPh(2))(NCMe)(3)]X(2) (4a, R = Ph; 4b, R = Cy; X(2) = FeBr(4) or (BF(4))(2)) form when 1 equiv of iron is reacted with PPh(2)CH(2)CH(2)NH(2) and 0.5 equiv of the appropriate phosphonium dimer. The evidence for P-N-P coordination is the large (2)J(PP) coupling constant in the (31)P {(1)H} NMR spectrum for the trans phosphorus nuclei. If 0.5 equiv of [Fe(H(2)O)(6)][BF(4)](2) were added in the synthesis, the complex trans [Fe(NCMe)(2)(Ph(2)PC(2)H(4)NH(2))(2)][FeBr(4)] (4c) formed, and this has been characterized by X-ray diffraction. Complexes [Fe{PR(2)CH(2)CH=NCH(2)(2 C(5)H(4)N)}(2)](BPh(4))(2) (5) are bis-tridentate iron(II) complexes with pyridyl donors trans to the phosphine donors. Interestingly, addition of the diamines ethylenediamine, (1R,2R)-(-)-1,2-diaminocyclohexane, (1R,2R)-(-)-1,2 diphenylethylenediamine, or o-phenylenediamine, in the template synthesis with 2 led directly to tetradentate P-N-N-P iron(II) complexes trans [Fe(NCMe)(2)(PCy(2)CH(2)CH=N-Q-N=CHCH(2)PCy(2)](BPh(4))(2) (Q = CH(2)CH(2), 6a; Q = (1R,2R)-cyclo-C(6)H(10), 6b; Q = (1R,2R)-CHPhCHPh, 6c; Q = C(6)H(4), 6d). In contrast, similar reactions under the same conditions with dimer 1 led to complexes mer-[Fe(P-N-N)(2)](2+) as reported previously. Complexes 6a and 6b have been characterized by X-ray diffraction and exhibited large P-Fe-P bond angles of 112.92(2) and 111.96(4) degrees, respectively. PMID- 20028111 TI - Comparison of microencapsulation properties of spruce galactoglucomannans and arabic gum using a model hydrophobic core compound. AB - In the present study, microencapsulation and the physical properties of spruce ( Picea abies ) Omicron-acetyl-galactoglucomannans (GGM) were investigated and compared to those of arabic gum (AG). Microcapsules were obtained by freeze drying oil-in-water emulsions containing 10 wt % capsule materials (AG, GGM, or a 1:1 mixture of GGM-AG) and 2 wt % alpha-tocopherol (a model hydrophobic core compound that oxidizes easily). Microcapsules were stored at relative humidity (RH) of 0, 33, and 66% at 25 degrees C for different time periods, and their alpha-tocopherol content was determined by HPLC. X-ray microtomography analyses showed that the freeze-dried emulsions of GGM had the highest and those of AG the lowest degree of porosity. According to X-ray diffraction patterns, both freeze dried AG and GGM showed an amorphous nature. The storage test showed that anhydrous AG microcapsules had higher alpha-tocopherol content than GGM containing capsules, whereas under 33 and 66% RH conditions GGM was superior in relation to the retention of alpha-tocopherol. The good protection ability of GGM was related to its ability to form thicker walls to microcapsules and better physical stability compared to AG. The glass transition temperature of AG was close to the storage temperature (25 degrees C) at RH of 66%, which explains the remarkable losses of alpha-tocopherol in the microcapsules under those conditions. PMID- 20028112 TI - Processing can alter the properties of peanut extract preparations. AB - As peanut allergy is an increasing public health risk, affecting over 1% of the United States and United Kingdom school children, it is important that methods and reagents for accurate diagnosis of food allergy and detection of allergenic foods are reliable and consistent. Given that most current experimental, diagnostic, and detection tests rely on the presence of soluble allergens in food extracts, we investigated the effects of thermal processing on the solubility and IgE binding of the major peanut allergens, Ara h 1 and Ara h 2. The soluble and insoluble fractions of peanuts that were boiled, fried, and roasted were subjected to electrophoresis and Western blot analysis using anti-Ara h 1 and anti-Ara h 2 antibodies and serum IgE from peanut allergic individuals. Overall protein solubility is reduced with processing and IgE binding increases in the insoluble fractions, due mostly to the increase in the amount of insoluble proteins, with increased time of heating in all processes tested. Therefore, it can be concluded that thermal processing of peanuts alters solubility, and the differences in protein solubility within various extract preparations may contribute to inconsistent skin prick test and immunoassay results, particularly when nonstandardized reagents are used. PMID- 20028113 TI - Thiol-capped ZnO nanowire/nanotube arrays with tunable magnetic properties at room temperature. AB - The present study reports room-temperature ferromagnetic behaviors in three dimensional (3D)-aligned thiol-capped single-crystalline ZnO nanowire (NW) and nanotube (NT) arrays as well as polycrystalline ZnO NT arrays. Besides the observation of height-dependent saturation magnetization, a much higher M(s) of 166 microemu cm(-2) has been found in NTs compared to NWs (36 microemu cm(-2)) due to larger surface area in ZnO NTs, indicating morphology-dependent magnetic properties in ZnO NW/NT systems. Density functional calculations have revealed that the origin of ferromagnetism is mainly attributed to spin-polarized 3p electrons in S sites and, therefore, has a strong correlation with Zn-S bond anisotropy. The preferential magnetization direction of both single-crystalline NTs and NWs lies perpendicular to the tube/wire axis due to the aligned high anisotropy orientation of the Zn-S bonds on the lateral (100) face of ZnO NWs and NTs. Polycrystalline ZnO NTs, however, exhibit a preferential magnetization direction parallel to the tube axis which is ascribed to shape anisotropy dominating the magnetic response. Our results demonstrate the interplay of morphology, dimensions, and crystallinity on spin alignment and magnetic anisotropy in a 3D semiconductor nanosystem with interfacial magnetism. PMID- 20028114 TI - Maskless nanoscale writing of nanoparticle-polymer composites and nanoparticle assemblies using thermal nanoprobes. AB - Nanoparticle polymer composites containing metal, semiconductor, magnetic, and optically active nanoparticles were deposited onto multiple substrates from a heatable atomic force microscope tip. The nanoparticle nanostructures were functional as deposited or could be etched with an oxygen plasma, revealing single nanoparticle lithographic resolution. Many types of nanoparticles can be patterned with the same technique, without the need to tailor the substrate chemistry and without solution processing. PMID- 20028115 TI - Thermoreversible covalent self-assembly of oligo(p-phenylenevinylene) bridged gold nanoparticles. AB - Organic-inorganic hybrids have been fabricated through mild Diels-Alder cross linking between maleimide bearing oligo(p-phenylenevinylene) (OPV) and furan functionalized gold nanoparticles with diameter smaller than 2 nm. The OPV ligands afford strong reaction ability toward furan group due to their maleimide moieties. These small gold nanoparticles form close-packed homogeneous hybrids with well-defined interfaces by incorporating OPV ligands in solutions. Covalent assembly and disassembly of gold nanoparticles can be achieved by repeated thermal stimuli on as-obtained hybrids, which can be monitored by fluorescence changes of OPVs and surface plasmon resonance absorption. Moreover, the dramatic photophysical properties and assembly behavior of these hybrids allow this procedure to be performed as a smart assay for monitoring the process of the Diels-Alder reaction. PMID- 20028116 TI - Feature-level MALDI-MS characterization of in situ-synthesized peptide microarrays. AB - Characterizing the chemical composition of microarray features is a difficult yet important task in the production of in situ-synthesized microarrays. Here, we describe a method to determine the chemical composition of microarray features, directly on the feature. This method utilizes nondiffusional chemical cleavage from the surface along with techniques from MALDI-MS tissue imaging, thereby making the chemical characterization of high-density microarray features simple, accurate, and amenable to high-throughput. PMID- 20028117 TI - Anthracene-bridged binuclear ruthenium complexes: electrochemical and spectroscopic evidence of electronic communication through the pi system. AB - Six dinuclear cyclometalated ruthenium complexes, 1-6, based on diphenylanthracene (DPA) and anthracene (AN) as bridging ligands have been synthesized and fully characterized electrochemically and spectroscopically. The anodic electrochemistry of the homobinuclear ruthenium complexes, 1-6, has been examined in three different nonaqueous solvents (ACN, DMF, and CH(2)Cl(2)). The ability of the anthracene derivatives to transmit electronic effects between the two redox units has been demonstrated by the observed splitting of the voltammetric signals ascribed to the metal centers. The electronic communication has also been evidenced by the presence of intervalence charge transfer transition bands in the near-infrared region of the spectrum due to an intramolecular electron transfer process mediated by the bridge when the mixed valence species (Ru(II)/Ru(III)) are electrochemically generated. Cyclic voltammetric measurements have been carried out under different conditions of solvent and supporting electrolyte. Differences in DeltaE degrees', the potential separation of the formal potentials of the metal-based anodic processes, have been observed and found to depend on the medium employed. These differences have been ascribed to different degrees of ion pairing. Such effects can be, in turn, modulated as a function of not only the polarity and donor strength of the solvent but also of the coordinating capacity of the anion employed as a supporting electrolyte. PMID- 20028118 TI - Rhodium-catalyzed asymmetric addition of arylboronic acids to beta phthaliminoacrylate esters toward the synthesis of beta-amino acids. AB - Rhodium-catalyzed asymmetric 1,4-addition of arylboronic acids to beta phthaliminoacrylate esters took place efficiently to give high yields of beta aryl-beta-amino acid esters with 96-99% enantioselectivity, which was realized by use of a hydroxorhodium/chiral diene complex. PMID- 20028119 TI - Electroosmotic flow can generate ion current rectification in nano- and micropores. AB - This paper introduces a strategy for generating ion current rectification through nano- and micropores. This method generates ion current rectification by electroosmotic-driven flow of liquids of varying viscosity (and hence varying conductance) into or out of the narrowest constriction of a pore. The magnitude of current rectification was described by a rectification factor, R(f), which is defined by the ratio of the current measured at a positive voltage divided by the current measured at a negative voltage. This method achieved rectification factors in the range of 5-15 using pores with diameters ranging from 10 nm to 2.2 microm. These R(f) values are similar to the rectification factors reported in other nanopore-based methods that did not employ segmented surface charges. Interestingly, this work showed that in cylindrical nanopores with diameters of 10 nm and a length of at least 275 nm, electroosmotic flow was present and could generate ion current rectification. Unlike previous methods for generating ion current rectification that require nanopores with diameters comparable to the Debye length, this work demonstrated ion current rectification in micropores with diameters 500 times larger than the Debye length. Thus this method extends the concept of fluidic diodes to the micropore range. Several experiments designed to alter or remove electroosmotic flow through the pore demonstrated that electroosmotic flow was required for the mode of ion current rectification reported here. Consequently, the magnitude of current rectification could be used to indicate the presence of electroosmotic flow and the breakdown of electroosmotic flow with decreasing ionic strength and hence increasing electric double layer overlap inside nanopores. PMID- 20028120 TI - Amine-induced Michael/Conia-ene cascade reaction: application to a formal synthesis of (+/-)-Clavukerin A. AB - An efficient and versatile amine-induced Michael/Conia-ene cascade cyclization reaction has been developed for establishing 6,6- and 5,7-bicyclic fused carbocycles with simple acyclic beta-ketoesters as the substrates in one-pot condition and this new cyclization method has been successfully utilized in a formal synthesis of (+/-)-Clavukerin A. PMID- 20028121 TI - UV Raman spectroscopic studies on active sites and synthesis mechanisms of transition metal-containing microporous and mesoporous materials. AB - Microporous and mesoporous materials are widely used as catalysts and catalyst supports. Although the incorporation of transition metal ions into the framework of these materials (by isomorphous substitution of Al and Si) is an effective means of creating novel catalytic activity, the characterization of the transition metal species within these materials is difficult. Both the low concentration of the highly dispersed transition metal and the coexistence of extraframework transition metal species present clear challenges. Moreover, the synthetic mechanisms that operate under the highly inhomogeneous conditions of hydrothermal synthesis are far from well understood. A useful technique for addressing these challenges is UV Raman spectroscopy, which is a powerful technique for catalyst characterization and particularly for transition metal containing microporous and mesoporous materials. Conventional Raman spectroscopy, using visible and IR wavelengths, often fails to provide the information needed for proper characterization as a result of fluorescence interference. But shifting the excitation source to the UV range addresses this difficulty: interference from fluorescence (which typically occurs at 300-700 nm or greater) is greatly diminished. Moreover, signal intensity is enhanced because Raman intensity is proportional to the fourth power of the scattered light frequency. In this Account, we review recent advances in UV Raman spectroscopic characterization of (i) highly dispersed transition metal oxides on supports, (ii) transition metal ions in the framework of microporous and mesoporous materials, and (iii) the synthetic mechanisms involved in making microporous materials. By taking advantage of the strong UV resonance Raman effect, researchers have made tremendous progress in the identification of isolated transition metal ions incorporated in the framework of microporous and mesoporous materials such as TS-1, Ti-MCM-41, Fe-ZSM-5, and Fe-SBA-15. The synthetic mechanisms involved in creating microporous materials (such as Fe-ZSM-5 and zeolite X) have been investigated with resonance and in situ UV Raman spectroscopy. The precursors and intermediates evolved in the synthesis solution and gels can be sensitively detected and followed during the course of zeolite synthesis. This work has resulted in a greater understanding of the structure of transition metal-containing microporous and mesoporous materials, providing a basis for the rational design and synthesis of microporous and mesoporous catalysts. PMID- 20028122 TI - Isolation and identification of triglycerides and ester oligomers from partial degradation of potato suberin. AB - Suberized cell walls from wound-healing potato tubers ( Solanum tuberosum ) were depolymerized under mild conditions using methanolic potassium hydroxide in order to investigate the chemical linkages present in this protective plant biopolymer. Analysis of the resulting soluble oligomeric fragments with HPLC, 1D and 2D NMR, LC/MS, and MS(n) methods allowed identification of several novel compounds: a family of homologous triglycerides, a family of homologous aliphatic ester trimers, and an ether-linked phenylacetic acid dimer. These findings illustrate the diversity of rigid and flexible molecular linkages present in both poly(aliphatic) and poly(aromatic) domains of potato suberin, and they point toward architectures that may account for its function as a potent hydrophobic barrier to water, thermal equilibration, and microbial pathogens. PMID- 20028123 TI - Differences in the molecular structure of beta(2)-microglobulin between two morphologically different amyloid fibrils. AB - Differences in the molecular structures of beta(2)-microglobulin between the two morphologically different amyloid fibrils having a needlelike [long-straight (LS)] and flexible [wormlike (WL)] character were investigated by infrared, Raman, and vacuum-ultraviolet circular dichroism spectroscopy. It turned out that although the beta-sheet content was comparable between the two kinds of fibrils (53 +/- 6% for the LS fibril and 47 +/- 6% for the WL fibril), the protonation states of the carboxyl side chains were distinctly different; the deprotonated (COO(-)) and protonated (COOH) forms were dominant in the LS and WL fibrils at pH 2.5, respectively, meaning that the pK(a) is specifically lowered in the LS fibril. Such a difference was not observed for the fibrils of the core fragments. Since site-specific interactions generally cause variation in the pK(a) of carboxyl side chains in proteins, these results suggest that "hook"-like interactions generated by hydrogen bonding and the formation of a salt bridge are present in the LS fibril, providing enthalpic stabilization. Presumably, the carboxyl groups fix the spatial arrangement of beta-strands and beta-sheets, bringing about the needlelike morphology. The absence of this regulation would result in the flexible morphology of the WL fibril, providing entropic stabilization. PMID- 20028124 TI - The ability of rodent islet amyloid polypeptide to inhibit amyloid formation by human islet amyloid polypeptide has important implications for the mechanism of amyloid formation and the design of inhibitors. AB - Islet amyloid polypeptide (IAPP) is a 37-residue polypeptide hormone that is responsible for islet amyloid formation in type II diabetes. Human IAPP is extremely amyloidogenic, while rat IAPP and mouse IAPP do not form amyloid in vitro or in vivo. Rat IAPP and mouse IAPP have identical primary sequences, but differ from the human polypeptide at six positions, five of which are localized between residues 20 and 29. The ability of rat IAPP to inhibit amyloid formation by human IAPP was tested, and the rat peptide was found to be an effective inhibitor. Thioflavin-T fluorescence-monitored kinetic experiments, transmission electron microscopy, and circular dichroism showed that rat IAPP lengthened the lag phase for amyloid formation by human IAPP, slowed the growth rate, reduced the amount of amyloid fibrils produced in a dose-dependent manner, and altered the morphology of the fibrils. The inhibition of human IAPP amyloid formation by rat IAPP can be rationalized by a model that postulates formation of an early helical intermediate during amyloid formation where the helical region is localized to the N-terminal region of IAPP. The model predicts that proline mutations in the putative helical region should lead to ineffective inhibitors as should mutations that alter the peptide-peptide interaction interface. We confirmed this by testing the ability of A13P and F15D point mutants of rat IAPP to inhibit amyloid formation by human IAPP. Both these mutants were noticeably less effective inhibitors than wild-type rat IAPP. The implications for inhibitor design are discussed. PMID- 20028126 TI - Fe(x)O(y)@C spheres as an excellent catalyst for Fischer-Tropsch synthesis. AB - We demonstrate a one-pot hydrothermal cohydrolysis-carbonization process using glucose and iron nitrate as starting materials for the fabrication of carbonaceous spheres embedded with iron oxide nanoparticles. It is verified by TEM, (57)Fe Mossbauer, and Fe K-edge XAS that iron oxide nanoparticles are highly dispersed in the carbonaceous spheres, leading to a unique microstructure. A formation mechanism is also proposed. This route is also applicable to a range of other naturally occurring saccharides and metal nitrates. A catalytic study revealed the remarkable stability and selectivity of the reduced Fe(x)O(y)@C spheres in the Fischer-Tropsch synthesis, which clearly exemplifies the promising application of such materials. PMID- 20028125 TI - Functional studies and modeling of pore-lining residue mutants of the influenza a virus M2 ion channel. AB - The A/M2 protein of influenza A virus forms a tetrameric proton-selective pH gated ion channel. The H(37)xxxW(41) motif located in the channel pore is responsible for its gating and proton selectivity. Channel activation most likely involves protonation of the H37 residues, while the conductive state of the channel is characterized by two or three charged His residues in a tetrad. A/M2 channel activity is inhibited by the antiviral drug amantadine. Although a large number of functional amantadine-resistant mutants of A/M2 have been observed in vitro, only a few are observed in highly transmissible viruses in the presence or absence of amantadine. We therefore examined 49 point mutants of the pore-lining residues, representing both natural and nonnatural variants. Their ion selectivity, amantadine sensitivity, specific activity, and pH-dependent conductance were measured in Xenopus oocytes. These measurements showed how variations in the sequence lead to variations in the proton conduction. The results are consistent with a multistep mechanism that allows the protein to fine tune its pH-rate profile over a wide range of proton concentrations, hypothesized to arise from different protonation states of the H37 tetrad. Mutations that give native-like conductance at low pH as well as minimal leakage current at pH 7.0 were surprisingly rare. Moreover, the results are consistent with a location of the amantadine-binding site inside the channel pore. These findings have helped to define the set of functionally fit mutants that should be targeted when considering the design of novel drugs that inhibit amantadine-resistant strains of influenza A virus. PMID- 20028127 TI - A tandem reaction initiated by 1,4-addition of bis(iodozincio)methane for 1,3 diketone formation. AB - Treatment of an gamma-acyloxy-alpha,beta-unsaturated ketone with bis(iodozincio)methane leads to a novel tandem reaction consisting of three steps: (1) 1,4-addition of the dizinc reagent to the enone, which affords the corresponding zinc enolate of the beta-zinciomethylated ketone; (2) intramolecular nucleophilic attack by the enolate on the ester group; and (3) Grob-type fragmentation of the adduct, accompanied by elimination of the zinc alkoxide of allyl alcohol. The overall reaction gives 1,3-diketones efficiently. PMID- 20028128 TI - Development and validation of a liquid chromatography tandem mass spectrometry method for simultaneous determination of four anthocyanins in human plasma after black currant anthocyanins ingestion. AB - Black currant anthocyanins consist of delphinidin-3-glucoside (1), delphinidin-3 rutinoside (2), cyanidin-3-glucoside (3), and cyanidin-3-rutinoside (4). A liquid chromatography tandem mass spectrometry method for simultaneous determination of four anthocyanins in human plasma was developed and validated. Samples were prepared using solid phase extraction, followed by chromatographic separation with a reverse phase C(18) column with gradient elution using mobile phases containing water, acetonitrile, and formic acid. The quantification of four anthocyanins was determined by multiple reaction monitoring using electrospray ionization. The method showed good selectivity, sensitivity (limits of quantification for four anthocyanins were 0.2 nmol/L), linearity (0.2-20 nmol/L; r > 0.999), intra- and interday precision, accuracy (<14%), and recovery (62.5 85.7%). Analyte stability was investigated in detail. This method was successfully applied to the determination of delphinidin-3-glucoside (1), delphinidin-3-rutinoside (2), cyanidin-3-glucoside (3), and cyanidin-3-rutinoside (4) concentrations in human plasma after ingestion of a single dose of black currant anthocyanins (87.9 micromol (58.8 mg) total anthocyanins). PMID- 20028129 TI - In vitro proteolytic inhibition, polyphenol oxidase activity, and soluble o diphenols in grasses and cereals. AB - Producing high quality forage remains a challenge due to potential protein degradation during ensiling. Ten grass species and four cereals were screened for proteolytic inhibition, polyphenol oxidase (PPO) activity, and the presence of soluble PPO substrates. In orchardgrass, ryegrass, smooth bromegrass, and meadow fescue extracts, the addition of caffeic acid resulted in 70-98% reduction in proteolysis for both greenhouse and field grown environments. These same grass extracts exhibited 10- to 1000-fold higher PPO activity compared to all other grasses and cereals, depending on the substrate supplied and environment grown. Chlorogenic acid and caffeic acid were the two abundant soluble o-diphenols. Tall fescue and timothy exhibiting low PPO activity contained the highest levels of chlorogenic acid (3-11 micromol g(-1) FW): 10- to 100-fold more than any o diphenol in all other grasses. These results suggest that several grass species contain PPO activity, but may lack appropriate o-diphenol substrates to effectively inhibit proteolysis during ensiling while other grasses lack PPO activity but contain potential soluble substrate. Environmental factors appear to influence these parameters, creating an opportunity to exploit and enhance proteolytic inhibition during ensiling by genetic manipulation. PMID- 20028130 TI - Dynamics of ultrafast singlet and triplet charge transfer in anthraquinone-DNA conjugates. AB - The efficiency of singlet and triplet charge radical ion-pair formation and the dynamics of radical-pair charge recombination in DNA-anthraquinone conjugates have been investigated by means of femtosecond time-resolved transient absorption spectroscopy. Singlet charge separation is more efficient than intersystem crossing, resulting in inefficient formation of the long-lived triplet radical ion pair. Both singlet charge separation and charge recombination are faster when guanine rather than adenine is the neighboring purine base. PMID- 20028131 TI - Degradation of sulfonamides in aqueous solution by membrane anodic fenton treatment. AB - Two agricultural antibiotics used heavily in agriculture, sulfamethazine and sulfadiazine, were degraded in an aqueous system by anodic Fenton treatment (AFT), an advanced oxidation technique that has been shown to be effective in degrading various pesticides but has not been applied to antibiotics. The effects of the H(2)O(2)/Fe(2+) ratio, Fe(2+) delivery rate, and initial contaminant concentration on the degradation of sulfamethazine by AFT were determined. The optimal H(2)O(2)/Fe(2+) ratio was determined to be 10:1, and the optimal Fe(2+) delivery rate was found to be between 38.9 and 54.4 microM min(-1). Under these conditions, sulfamethazine was completely degraded within 10 min at a range of concentrations (18-250 microM) commonly found in manure lagoons, contaminated rivers, and groundwater. Using the same optimal conditions, the effect of pH on the degradation of sulfadiazine by AFT was analyzed, and 100 microM sulfadiazine was degraded within 6-8 min of treatment at a range of pH values (3.1-7.1) that could potentially be found in aquatic environments. Degradation products and pathways were proposed for both compounds, and it was inferred that AFT degradation products of sulfadiazine and sulfamethazine are unlikely to retain the bacteriostatic properties of their parent compounds. An aquatic toxicity test employing Lemna gibba confirmed that AFT removes the bacteriostatic properties of sulfamethazine and sulfadiazine during degradation. PMID- 20028132 TI - Dynamic kinetic resolution: asymmetric transfer hydrogenation of alpha-alkyl substituted beta-ketoamides. AB - Dynamic kinetic resolution (deracemization) of various alpha-alkyl-substituted beta-ketoamides 1 via asymmetric transfer hydrogenation proceeded efficiently to give the corresponding syn-beta-hydroxy amides 3 in high diastereo- and enantioselectivities. Specifically, subjection of 1 to HCO(2)H and Et(3)N in the presence of 0.5-1 mol % of pentafluorobenzenesulfonyl-DPEN-Ru catalyst 2b at 30 40 degrees C in either PhCH(3) or CH(2)Cl(2) generated the syn-hydroxy product 3 selectively in 15-33:1 dr, 93-97% ee, and 75-88% isolated yields. PMID- 20028133 TI - Cloning and molecular analysis of HlbZip1 and HlbZip2 transcription factors putatively involved in the regulation of the lupulin metabolome in hop (Humulus lupulus L.). AB - Hop (Humulus lupulus L.), the essential source of beer flavor is of interest from a medicinal perspective in view of its high content in health-beneficial terpenophenolics including prenylflavonoids. The dissection of biosynthetic pathway(s) of these compounds in lupulin glands, as well as its regulation by transcription factors (TFs), is important for efficient biotechnological manipulation of the hop metabolome. TFs of the bZIP class were preselected from the hop transcriptome using a cDNA-AFLP approach and cloned from a cDNA library based on glandular tissue-enriched hop cones. The cloned TFs HlbZIP1A and HlbZIP2 have predicted molecular masses of 27.4 and 34.2 kDa, respectively, and both are similar to the group A3 bZIP TFs according to the composition of characteristic domains. While HlbZIP1A is rather neutral (pI 6.42), HlbZIP2 is strongly basic (pI 8.51). A truncated variant of HlbZIP1 (HlbZIP1B), which is strongly basic but lacks the leucine zipper domain, has also been cloned from hop. Similar to the previously cloned HlMyb3 from hop, both bZIP TFs show a highly specific expression in lupulin glands, although low expression was observed also in other tissues including roots and immature pollen. Comparative functional analyses of HlbZip1A, HlbZip2, and subvariants of HlMyb3 were performed in a transient expression system using Nicotiana benthamiana leaf coinfiltration with Agrobacterium tumefaciens strains bearing hop TFs and selected promoters fused to the GUS reference gene. Both hop bZIP TFs and HlMyb3 mainly activated the promoters of chalcone synthase chs_H1 and the newly cloned O-methyl transferase 1 genes, while the response of the valerophenone synthase promoter to the cloned hop TFs was very low. These analyses also showed that the cloned bZIP TFs are not strictly G-box-specific. HPLC analysis of secondary metabolites in infiltrated Petunia hybrida showed that both hop bZIP TFs interfere with the accumulation and the composition of flavonol glycosides, phenolic acids, and anthocyanins, suggesting the possibility of coregulating flavonoid biosynthetic pathways in hop glandular tissue. PMID- 20028134 TI - Bafilomycins produced in culture by Streptomyces spp. isolated from marine habitats are potent inhibitors of autophagy. AB - Five new bafilomycins, F (1) to J (5), have been isolated from laboratory cultures of two Streptomyces spp. obtained from marine sediments collected in British Columbia, and their structures have been elucidated by detailed analysis of spectroscopic data and the synthesis of model compounds. The new bafilomycins F (1), G (2), H (3), and J (5) along with several co-occurring known analogues showed potent inhibition of autophagy in microscopy and biochemical assays. The thiomorpholinone fragment present in bafilomycin F (1) has not previously been found in a natural product. PMID- 20028135 TI - Air-stable operation of transparent, colloidal quantum dot based LEDs with a unipolar device architecture. AB - We report a novel unipolar light-emitting device architecture that operates using direct-current, field-driven electroluminescence of colloidally synthesized quantum dots (QDs). This device architecture, which is based only on transparent ceramics and QDs, enables emission from different color QDs and, for the first time, constant QD electroluminescence during extended operation in air, unpackaged. PMID- 20028136 TI - Phosphoproteome analysis of rat L6 myotubes using reversed-phase C18 prefractionation and titanium dioxide enrichment. AB - The rat L6 myotubes is an important in vitro model system for studying signaling pathways in skeletal muscle. Exploring phosphorylation events involved in the skeletal muscle is very significant for elucidating the kinase-substrate relationship, understanding regulatory mechanisms involved in signaling pathways and providing insights into numerous cell processes. Here, we used mass spectrometry-based proteomics to conduct global phosphoproteome profiling of rat L6 myotubes. Using an efficient phosphoproteomic strategy including prefractionation of tryptic peptide mixtures with self-packed RP C18 columns, phosphopeptide enrichment with TiO(2), and 2D-LC (SCX/RP)-MS/MS analysis, a total of 2230 unique phosphopeptides from 1195 proteins were identified with a false discovery rate of less than 1.0% using a target/decoy database searching strategy. After determining the degree of certainty of the phosphorylation site location (Ascore value >or=19), 11 Ser motifs and one Thr motif were derived from our data set using the Motif-X algorithm. Several potential signaling pathways were found in our myotubes phosphoproteome, such as the MAPK signaling pathway and the IGF-1/Insulin signaling pathway. PMID- 20028137 TI - Identification of the binding site for the regulatory calcium-binding domain in the catalytic domain of NOX5. AB - NADPH oxidases (NOX) are important superoxide producing enzymes that regulate a variety of physiological and pathological processes such as bacteria killing, angiogenesis, sperm-oocyte fusion, and oxygen sensing. NOX5 is a member of the NOX family but distinct from the others by the fact that it contains a long N terminus with four EF-hand Ca(2+)-binding sites (NOX5-EF). NOX5 generates superoxide in response to intracellular Ca(2+) elevation in vivo and in a cell free system. Previously, we have shown that the regulatory N-terminal EF-hand domain interacts directly and in a Ca(2+)-dependent manner with the catalytic C terminal catalytic dehydrogenase domain (CDHD) of the enzyme, leading to its activation. Here we have characterized the interaction site for the regulatory NOX5-EF in the catalytic CDHD of NOX5 using cloned fragments and synthetic peptides of the CDHD. The interaction was monitored with pull-down techniques, cross-linking experiments, tryptophan fluorescence, hydrophobic exposure, isothermal titration calorimetry, and cell-free system enzymatic assays. This site is composed of two short segments: the 637-660 segment, referred to as the regulatory EF-hand-binding domain (REFBD), and the 489-505 segment, previously identified as the phosphorylation region (PhosR). NOX5-EF binds to these two segments in a Ca(2+)-dependent way, and the superoxide generation by NOX5 depends on this interaction. Controlled proteolysis suggests that the REFBD is autoinhibitory and inhibition is relieved by NOX5-EF. PMID- 20028138 TI - Desiccation-induced structuralization and glass formation of group 3 late embryogenesis abundant protein model peptides. AB - Anhydrobiotic (i.e., life without water) organisms are known to produce group 3 late embryogenesis abundant (G3LEA) proteins during adaptation to severely water deficient conditions. Their primary amino acid sequences are composed largely of loosely conserved 11-mer repeat units. However, little information has been obtained for the structural and functional roles of these repeat units. In this study, we first explore the consensus sequences of the 11-mer repeat units for several native G3LEA proteins originating from anhydrobiotic organisms among insects (Polypedilum vanderplanki), nematodes, and plants. Next, we synthesize four kinds of model peptides (LEA models), each of which consists of four or two repeats of the 11-mer consensus sequences for each of the three organisms. The structural and thermodynamic properties of the LEA models were examined in solution, in dehydrated and rehydrated states, and furthermore in the presence of trehalose, since a great quantity of this sugar is known to be produced in the dried cells of most anhydrobiotic organisms. The results of Fourier transform infrared (FTIR) spectroscopic measurements indicate that all of the LEA models transform from random coils to alpha-helical coiled coils on dehydration and return to random coils again on rehydration, both with and without trehalose. In contrast, such structural changes were never observed for a control peptide with a randomized amino acid sequence. Furthermore, our differential scanning calorimetry (DSC) measurements provide the first evidence that the above 11-mer motif-containing peptides themselves vitrify with a high glass transition temperature (>100 degrees C) and a low enthalpy relaxation rate. In addition, they play a role in reinforcing the glassy matrix of the coexisting trehalose. On the basis of these results, we discuss the underlying mechanism of G3LEA proteins as desiccation stress protectants. PMID- 20028139 TI - Mechanism of interaction between single-stranded DNA binding protein and DNA. AB - A single-stranded DNA binding protein (SSB), labeled with a fluorophore, interacts with single-stranded DNA (ssDNA), giving a 6-fold increase in fluorescence. The labeled protein is the adduct of the G26C mutant of the homotetrameric SSB from Escherichia coli and a diethylaminocoumarin {N-[2 (iodoacetamido)ethyl]-7-diethylaminocoumarin-3-carboxamide}. This adduct can be used to assay production of ssDNA during separation of double-stranded DNA by helicases. To use this probe effectively, as well as to investigate the interaction between ssDNA and SSB, the fluorescent SSB has been used to develop the kinetic mechanism by which the protein and ssDNA associate and dissociate. Under conditions where approximately 70 base lengths of ssDNA wrap around the tetramer, initial association is relatively simple and rapid, possibly diffusion controlled. The kinetics are similar for a 70-base length of ssDNA, which binds one tetramer, and poly(dT), which could bind several. Under some conditions (high SSB and/or low ionic strength), a second tetramer binds to each 70-base length, but at a rate 2 orders of magnitude slower than the rate of binding of the first tetramer. Dissociation kinetics are complex and greatly accelerated by the presence of free wild-type SSB. The main route of dissociation of the fluorescent SSB x ssDNA complex is via association first with an additional SSB and then dissociation. Comparison of binding data with different lengths of ssDNA gave no evidence of cooperativity between tetramers. Analytical ultracentrifugation was used to determine the dissociation constant for labeled SSB(2) x dT(70) to be 1.1 microM at a high ionic strength (200 mM NaCl). Shorter lengths of ssDNA were tested for binding: only when the length is reduced to 20 bases is the affinity significantly reduced. PMID- 20028140 TI - Characterization of the DNA binding activity of the ZFY zinc finger domain. AB - The ZFY protein is a member of one of the most interesting classes of polydactyl zinc finger proteins. It has a domain of 13 tandem zinc fingers that is organized with an internal repeat of odd-even finger pairs. It has been proposed that each finger pair interacts with six base pairs within a turn of the double helix, the downstream linker crossing the minor groove to place the next finger pair on the following turn of the DNA. Yet putative binding sites for the full-length ZFY protein appear to consist of a six-base AGGCCY consensus sequence that is present in one or two copies. In this study the equilibrium binding of two ZFY-derived zinc finger peptides to 4R DNA with tandem copies of the consensus sequence was investigated. The ZFY5 peptide contains fingers 5-13, including four odd-even finger pairs, and the ZFY11 peptide contains fingers 11-13 and has one odd-even finger pair. Both peptides bound to 4R DNA with equal affinities, forming a bimolecular complex that is mediated by the downstream AGGCCY motif. The additional odd-even finger pairs in ZFY5 made no measurable difference in the mechanism of DNA binding compared to ZFY11. The effects on the DNA-protein interaction of mutations in the 4R DNA and in the key alpha-helical residues of fingers 11-13 indicate that the binding of ZFY to DNA is mediated by the interaction of the GGCC core base pairs with fingers 12 and 13. These results demonstrate that the even-odd repeats in the ZFY zinc finger domain do not make significant contributions to DNA binding. PMID- 20028141 TI - Free-energy landscape of RNA hairpins constructed via dihedral angle principal component analysis. AB - To systematically construct a low-dimensional free-energy landscape of RNA systems from a classical molecular dynamics simulation, various versions of the principal component analysis (PCA) are compared: the cPCA using the Cartesian coordinates of all atoms, the dPCA using the sine/cosine-transformed six backbone dihedral angles as well as the glycosidic torsional angle chi and the pseudorotational angle P, the aPCA which ignores the circularity of the 6 + 2 dihedral angles of the RNA, and the dPCA(etatheta), which approximates the 6 backbone dihedral angles by 2 pseudotorsional angles eta and theta. As representative examples, a 10-nucleotide UUCG hairpin and the 36-nucleotide segment SL1 of the Psi site of HIV-1 are studied by classical molecular dynamics simulation, using the Amber all-atom force field and explicit solvent. It is shown that the conformational heterogeneity of the RNA hairpins can only be resolved by an angular PCA such as the dPCA but not by the cPCA using Cartesian coordinates. Apart from possible artifacts due to the coupling of overall and internal motion, this is because the details of hydrogen bonding and stacking interactions but also of global structural rearrangements of the RNA are better discriminated by dihedral angles. In line with recent experiments, it is found that the free energy landscape of RNA hairpins is quite rugged and contains various metastable conformational states which may serve as an intermediate for unfolding. PMID- 20028142 TI - Proton-regulated electron transfers from tyrosine to tryptophan in proteins: through-bond mechanism versus long-range hopping mechanism. AB - Charge transfer between tyrosine and tryptophan residues in proteins is continuously a hot topic because of its important biological implication. On the basis of DFT calculations and ab initio molecular dynamics simulations, several possible proton/electron cooperative transfer mechanisms from tyrosine to a tryptophan radical (or cation) without or with the assistence of a base in proteins are proposed in this work which range from direct proton-coupled pi electron pi-channel/sigma-channel transfers (PC(pi)E(pi)T versus PC(pi)E(sigma)T) to proton-coupled long-range hopping mechanisms depending on the peptide conformations. In general, because of a smaller ionization potential, tryptophan readily behaves as two oxidized states: dehydrogenated neutral radical and ionized radical cation. For the neutral radical, the proton/electron transfers between a tyrosine and a tryptophan radical prefer a cooperative direct coupling mode through the PC(pi)E(pi)T or PC(pi)E(sigma)T mechanism if two residues are proximal or can approach each other, while they cannot take place without assistence if two residues are far apart. The tryptophan radical cation prefers to form a complex with tyrosine to stabilize the hole when two residues are proximal or can approach each other. However, the electron transfer from tyrosine to tryptophan could occur via a hopping mechanism but is regulated by a base as a proton acceptor in the vicinity of tyrosine when two residues are separated. The dynamics properties and characters for these transfer events are also presented. The energetics comparison indicates that the energy barriers for the direct PCET (PC(pi)E(pi)T or PC(pi)E(sigma)T) mechanisms are higher than those of the base assisting hopping mechanism (6.8-12.5 versus OH + Cl reaction at a 0.26 eV collision energy: a comparison between theory and experiment. AB - The dynamics of the O((1)D) + HCl(v = 0, j = 0) --> Cl + OH reaction at a 0.26 eV collision energy has been investigated by means of a quasiclassical trajectory (QCT) and statistical quantum and quasiclassical methods. State-resolved cross sections and Cl atom velocity distributions have been calculated on two different potential energy surfaces (PESs): the H2 surface (Martinez et al. Phys. Chem. Chem. Phys. 2000, 2, 589) and the latest surface by Peterson, Bowman, and co workers (PSB2) (J. Chem. Phys. 2000, 113, 6186). The comparison with recent experimental results reveals that the PSB2 PES manages to describe correctly differential cross sections and the velocity distributions of the departing Cl atom. The calculations on the H2 PES seem to overestimate the OH scattering in the forward direction and the fraction of Cl at high recoil velocities. Although the comparison of the corresponding angular distributions is not bad, significant deviations with a statistical description are found, thus ruling out a complex forming mechanism as the dominant reaction pathway. However, for the ClO + H product channel, the QCT and statistical predictions are found to be in good agreement. PMID- 20028156 TI - Electron transfer collisions with oriented trifluoroacetic acid (CF3CO2H). AB - Electron transfer collisions between neutral K atoms and neutral, oriented trifluoroacetic acid molecules, CF(3)CO(2)H, are studied in crossed molecular beams at center of mass energies from 6 to 18 eV. An electron transfer produces a pair of ions with enough energy to escape the Coulomb attraction, and the ions are detected in separate time-of-flight mass spectrometers. The principle ions formed are K(+) and the trifluoroacetate ion, CF(3)CO(2)(-) ion, and this channel is favored for attack at the positive (-CO(2)H) end of the molecule. The steric asymmetry suggests that the electron is transferred into the pi*(CO) orbital. The nascent K(+) perturbs the molecular symmetry, allowing electron migration to the sigma*(OH) orbital to break the O-H bond and form CF(3)CO(2)(-). PMID- 20028157 TI - Hydrated complexes of atmospheric interest: rotational spectrum of diacetyl water. AB - The rotational spectrum of the molecular complex diacetyl-water has been measured by pulsed jet Fourier transform microwave (FTMW) spectroscopy. The water molecule acts as an electrofore and induces a dipole moment when combined with nonpolar diacetyl. Only one conformer has been observed, corresponding to the global minimum. Its shape is unambiguously established: the water moiety is linked asymmetrically to the organic molecule; it behaves as a proton donor to one of its oxygen atoms and interferes with the internal rotation of adjacent methyl group through a C-H...O interaction. Each rotational transition appears as a quintuplet, due to the internal rotation of the two methyl groups, which are nonequivalent in the adduct. From the analysis of the observed splittings, the V(3) barriers to the internal rotation of the two methyl groups have been determined to be 3.81(2) and 4.11(2) kJ/mol, respectively. PMID- 20028158 TI - Effect of the total angular momentum on the dynamics of the H2 + H2 system. AB - Extended full-dimensional quasiclassical trajectory calculations have been performed for the H(a)H(b) (v(ab) = 10, 11, 12, 13, 14, j(ab) = 0) + H(c)H(d) (v(cd) = 0, j(cd) = 0) collisions at values of the translational energy ranging from threshold to 1.5 eV and values of the total angular momentum quantum number J varying from zero to very large ones. Collision-induced dissociation, four center exchange reaction, and single exchange process probabilities have been calculated. Full-dimensional classical calculations were found to reproduce well the corresponding (J = 0) quantum results, including the thresholds. In contrast, the agreement of full-dimensional classical calculations with the corresponding both quantum and classical reduced dimensionality ones was found to be poor. The effect of varying J on the efficiency of the various processes has also been investigated. Four-center reactions were found to be favored by low values of J, whereas dissociation processes were found to be favored by higher values of J, as expected from the fact that energy exchange takes place at longer range than mass exchange. To evaluate to what extent the J = 0 full-dimensional calculations represent the unconstrained dynamics of the system, J-shift model classical results were compared with the all-J ones. Product vibrational distributions for both partially dissociative and exchange processes were also found to depend significantly on the value of J. PMID- 20028159 TI - A computational investigation of the multiple channels of the NF2 + F reaction. AB - We have theoretically studied the NF(3) = NF(2) + F, NF(2) + F = NF + F(2), and NF(2) + F = NF(2) + F reactive processes. More precisely, we have evaluated the thermal rate constants (TRC), with the Wigner and Eckart tunneling corrections, minimum energy path, and the intrinsic reaction coordinates of these systems. The NF(3) = NF(2) + F conventional and Wigner TRCs agree very well with experimental data available in the literature for a wide range of temperatures. This study gives a first step to understand and determine the correct decomposition path of nitrogen trifluoride (NF(3)). PMID- 20028160 TI - Phase space theory of evaporation in neon clusters: the role of quantum effects. AB - Unimolecular evaporation of neon clusters containing between 14 and 148 atoms is theoretically investigated in the framework of phase space theory. Quantum effects are incorporated in the vibrational densities of states, which include both zero-point and anharmonic contributions, and in the possible tunneling through the centrifugal barrier. The evaporation rates, kinetic energy released, and product angular momentum are calculated as a function of excess energy or temperature in the parent cluster and compared to the classical results. Quantum fluctuations are found to generally increase both the kinetic energy released and the angular momentum of the product, but the effects on the rate constants depend nontrivially on the excess energy. These results are interpreted as due to the very few vibrational states available in the product cluster when described quantum mechanically. Because delocalization also leads to much narrower thermal energy distributions, the variations of evaporation observables as a function of canonical temperature appear much less marked than in the microcanonical ensemble. While quantum effects tend to smooth the caloric curve in the product cluster, the melting phase change clearly keeps a signature on these observables. The microcanonical temperature extracted from fitting the kinetic energy released distribution using an improved Arrhenius form further suggests a backbending in the quantum Ne(13) cluster that is absent in the classical system. Finally, in contrast to delocalization effects, quantum tunneling through the centrifugal barrier does not play any appreciable role on the evaporation kinetics of these rather heavy clusters. PMID- 20028161 TI - Silylene defect at the dihydrogen terminated (100) Si surface. AB - Density functional calculations for both periodic slabs and different size cluster models of the hydrogen-terminated (100) surface of silicon are used to study a new configuration, formed by a silylene center interacting with vicinal silicon dihydrides through nonconventional hydrogen bonds. A comparison between slab-model and cluster-model approaches to modeling surface silylene defect formation processes is presented. The cluster models are used to analyze the structure and bonding of the silylene with a Lewis acid and base, showing the Zwitterionic nature of the defect. The silylene is also demonstrated to behave as a strong Bronsted acid. The stabilization of the silylene defect via interaction with species unavoidably present in the HF(aq)-etching solution is investigated. Finally, the negative chemical shift observed by X-ray photoelectron spectroscopy in the HF(aq)-etched (100) Si surface is attributed to the occurrence of silylene defect. PMID- 20028162 TI - Induced orbital paramagnetism and paratropism in closed-shell molecules. AB - Three-dimensional models of the quantum-mechanical current density induced by a uniform magnetic field in the electron cloud have been obtained for closed-shell systems BeH(-), BH, and CH(+), characterized by induced orbital paramagnetism, and in planar unsaturated hydrocarbons C(4)H(4) and clamped C(8)H(8), exhibiting pi paramagnetism. It is shown that, even for these paramagnetic systems, the paramagnetic contributions to magnetic susceptibilities and nuclear magnetic shielding, customarily taken into account in perturbation theory approaches, can formally be eliminated via the procedure of continuous transformation of the origin of the current density-paramagnetic zero. The definition of magnetic response properties can therefore be recast as a sum of two formally "diamagnetic" terms for any molecule, including systems showing strong induced orbital paramagnetism. It is shown that the paramagnetism in the compounds studied arises from the nodal topology of the electronic wave function. In particular, paratropic vortices circulate about stagnation lines at the intersection of nodal surfaces of the highest-occupied zero-order molecular orbital and corresponding first-order orbital. PMID- 20028163 TI - Differential cross sections and product rotational polarization in A + BC reactions using wave packet methods: H+ + D2 and Li + HF examples. AB - The state-to-state differential cross sections for some atom + diatom reactions have been calculated using a new wave packet code, MAD-WAVE3, which is described in some detail and uses either reactant or product Jacobi coordinates along the propagation. In order to show the accuracy and efficiency of the coordinate transformation required when using reactant Jacobi coordinates, as recently proposed [ J. Chem. Phys. 2006 , 125 , 054102 ], the method is first applied to the H + D(2) reaction as a benchmark, for which exact time-independent calculations are also performed. It is found that the use of reactant coordinates yields accurate results, with a computational effort slightly lower than that when using product coordinates. The H(+) + D(2) reaction, with the same masses but a much deeper insertion well, is also studied and exhibits a completely different mechanism, a complex-forming one which can be treated by statistical methods. Due to the longer range of the potential, product Jacobi coordinates are more efficient in this case. Differential cross sections for individual final rotational states of the products are obtained based on exact dynamical calculations for some selected total angular momenta, combined with the random phase approximation to save the high computational time required to calculate all partial waves with very long propagations. The results obtained are in excellent agreement with available exact time-independent calculations. Finally, the method is applied to the Li + HF system for which reactant coordinates are very well suited, and quantum differential cross sections are not available. The results are compared with recent quasiclassical simulations and experimental results [J. Chem. Phys. 2005, 122, 244304]. Furthermore, the polarization of the product angular momenta is also analyzed as a function of the scattering angle. PMID- 20028164 TI - Typicality in ensembles of quantum states: Monte Carlo sampling versus analytical approximations. AB - Random quantum states are presently of interest in the fields of quantum information theory and quantum chaos. Moreover, a detailed study of their properties can shed light on some foundational issues of the quantum statistical mechanics such as the emergence of well-defined thermal properties from the pure quantum mechanical description of large many-body systems. When dealing with an ensemble of pure quantum states, two questions naturally arise, what is the probability density function on the parameters which specify the state of the system in a given ensemble, and does there exist a "most typical" value of a function of interest in the considered ensemble? Here, two different ensembles are considered, the random pure state ensemble (RPSE) and the fixed expectation energy ensemble (FEEE). By means of a suitable parametrization of the wave function in terms of populations and phases, we focus on the probability distribution of the populations in these ensembles. A comparison is made between the distribution induced by the inherent geometry of the Hilbert Space and an approximate distribution derived by means of the minimization of the informational functional. While the latter can be analytically handled, the exact geometrical distribution is sampled by a Metropolis-Hastings algorithm. The analysis is made for an ensemble of wave functions describing an ideal system composed of n spins of 1/2 and reveals the salient differences between the geometrical and the approximate distributions. The analytical approximations are proven to be useful tools in order to obtain an ensemble-averaged quantity. In particular, we focus on the distribution of the Shannon entropy by providing an explanation of the emergence of a typical value of this quantity in the ensembles. PMID- 20028165 TI - Competition between photoisomerization and photocyclization of the cis isomers of n-styrylnaphthalenes and -phenanthrenes. AB - The isomerization and cyclization photoreactions of the cis (Z) isomers of n styrylnaphthalenes (n = 1 and 2), n-styrylphenanthrenes (n = 1, 2, 3, 4, and 9), and two related compounds, 3-styrylchrysene and 3-styrylbenzo[c]phenanthrene, were investigated by spectrophotometric and chromatographic techniques. The quantum yields of the two photoreactions were measured in aerated and deaerated nonpolar solvent at room temperature and compared with those reported in the literature for some of the molecules investigated and other related compounds. The combined use of high-performance liquid chromatographic and spectrophotometric techniques made easier the separation of the components of the photoreaction mixtures thus simplifying the study of the isomerization/cyclization competition and the measurement of the UV-vis absorption spectra and the thermal decay kinetics of the dihydrophenanthrene-type intermediates. The conformational equilibria in the ground state and the positional isomerism (n values) notably affect the prevalence of one or the other competitive photoreaction. Oxygen also plays an important role: practically it does not affect the quantum yield of the Z --> E process but has a drastic effect on the formation of the final oxidation product which can proceed even in the presence of traces of air. In three cases, non-negligible formation of a side colorless product was detected. It was assigned to an isomer of the normal photocyclization intermediate (4a,4b-dihydrophenanthrene-type), formed by shift of one or both hydrogen atom(s) to other positions of the rings. Parallel ab initio calculations of the potential energy of the possible conformers helped to explain the structure effects on the competitive photoreactive relaxation pathways. Some correlations between the computed reagent/product energy difference in the ground state and the photocyclization yield and thermal stability of the intermediates were also verified. PMID- 20028166 TI - Dissociation rate constant of the hydrogen fluoride dimer by the ab initio anharmonic RRKM theory. AB - The dissociation rate constants for hydrogen fluoride dimers are calculated using the YL method, proposed by Yao and Lin (Yao, L.; Mebel, A. M.; Lu, H. F.; Neusser, H. J.; Lin, S. H. J. Phys. Chem. A 2007, 111, 6722). The dividing surface method and RRKM theory are also used to obtain pseudo-transition states and rate constants, respectively. For the dissociation of HF dimers, the anharmonic rate constants are around 3.02 x 10(10) to 3.46 x 10(12) s(-1), while the harmonic rate constants are in the range of 2.93 x 10(10) to 1.66 x 10(13) s( 1), at a temperature range of 243-1000 K, for the canonical case. For the microcanonical case, the anharmonic rate constants are in the range of 1.91 x 10(12) to 1.32 x 10(13) s(-1) and the harmonic rate constants are in the range of 1.89 x 10(12) to 4.93 x 10(13) s(-1), with a total energy range of 1338-4500 cm( 1). Both harmonic and anharmonic rate constants are only comparable to the experimental results 0.5 x 10(10) s(-1) and 1 x 10(11) s(-1) in an inert gas bath at room temperature. In this case the harmonic and anharmonic rate constants show similar results in this calculation. The results of this paper indicate that the YL method is suitable for calculating dissociation rate constants of small flexible HF dimers and the anharmonic effect should be included. PMID- 20028167 TI - Calculation of MP2 and coupled-cluster molecular properties using the q-integral method. AB - The main purpose of this paper is to report results of quantum mechanical calculation of the H(2) system using the q-Integral method with correlation corrections to the SCF (Self Consistent Field) wave functions included through the Moller-Plesset second-order perturbation (MP(2)) and Coupled-Cluster (CC) theory. Using the q-Integral method, we evaluated potential energy curves, rovibrational spectroscopy constants, rovibrational spectra, interatomic equilibrium distance and longitudinal static hyper(polarizability). All calculations were carried out through the STO-3G, STO-6G, and double-zeta (DZV) atomic basis set. The q-Integral method was implemented in the source code of the general ab initio quantum chemistry package GAMESS. PMID- 20028168 TI - Activation of ethane C-H and C-C bonds by gas phase Th+ and U+: a theoretical study. AB - Two different approaches of density functional theory were used to analyze the C H and C-C bond activation mechanisms during the reaction of bare Th(+) and U(+) ions with ethane. We report a complete exploration of the potential energy surfaces taking into consideration different spin states. According to B3LYP/SDD computations the double dehydrogenation of C(2)H(6) is thermodynamically favorable only in the case of Th(+). It is shown that the overall C-H and C-C bond activation processes are exothermic in the case of Th(+) and endothermic for U(+). In both cases, the C-C insertion transition state barrier exceeds the energy of the ground state reactants, preventing the observation of these species under thermal conditions. PMID- 20028169 TI - Steric effects in the scattering of oriented CH3Cl molecular beam from a graphite surface: weak interaction of physisorption. AB - We report results of a study on steric effects appearing in the scattering of an oriented CH(3)Cl molecular beam from highly oriented pyrolytic graphite (HOPG) surface at 300 K. Data presented here show that the scattered CH(3)Cl beam intensity measured at a fixed scattering angle clearly depends on the initial molecular orientation toward the HOPG surface. The scattered CH(3)Cl beam intensity for the CH(3)-end collision is larger than that for the Cl-end collision, suggesting that strong anisotropy of the interaction potential induces the molecular-orientation-dependent energy dissipation during transient trapping into the physisorption well. PMID- 20028170 TI - Energy and density analyses of the 1Sigma(u)+ states in the H2 molecule from the united atom to dissociation. AB - The 1Sigma(u)+ excited states of the H2 molecule are computed following a recent study by Corongiu and Clementi (J. Chem. Phys. 2009, 131, 034301) on the 1Sigma(g)+ states. Full configuration interaction computations both from Hartree Fock molecular orbitals and Heitler-London atomic orbitals are presented and correlated with a comprehensive analysis. The basis sets utilized are either extended and optimized Slater type functions, STO, or spherical Gaussian functions, GTO. Computations and analyses are presented for states 1 to 14, covering the internuclear distances from 0.01 to 10000 bohr. The accurate data by L. Wolniewicz and collaborators, available for the first six excited states, verify the good quality of our computations. We focus on the characterization of the orbitals in the excited state wave functions, on the electronic density evolution from the united atom to dissociation, on quantitative decomposition of the total energy into covalent and ionic components and on detailed analyses of energy contributions to the total state energy from selected STO subsets. Each manifold has one state, specifically the states 1, 3 and 6, where the second minimum has strong ionic character. State 10 dissociates into the ion pair H+H-. PMID- 20028171 TI - Quantum Zeno effect in a model multilevel molecule. AB - We study the dynamics of the populations of a model molecule endowed with two sets of rotational levels of different parity, whose ground levels are energetically degenerate and coupled by a constant interaction. The relaxation rate from one set of levels to the other one has an interesting dependence on the average collision frequency of the molecules in the gas. This is interpreted as a quantum Zeno effect due to the decoherence effects provoked by the molecular collisions. PMID- 20028172 TI - Photophysical properties of 1,3,5-tris(2-naphthyl)benzene and related less arylated compounds: experimental and theoretical investigations. AB - Recently, a growing interest has concerned compounds characterized by high chemical and photophysical stability and high quantum yield for their possible technological applications. 1,3,5-Tris(2-naphthyl)benzene (N3B), 1,3-bis(2 naphthyl)benzene (N2B), and 2-naphthyl-benzene (N1B) are promising compounds, but they needed a detailed photophysical characterization. In this context, theoretical and experimental investigations have been carried out. Steady-state and decay time fluorescence measurements indicate that the second naphthyl group, added in the meta position of N1B, perturbs the electronic levels, whereas the further naphthyl addition, leading to N3B, does not promote changes in all of the observed properties. The investigated compounds show a biexponential fluorescence decay that has been attributed to a rearrangement involving the exited states S(1) and S(2). The minimum structure corresponding to the S(1) and S(2) states has been obtained at the configuration interaction with single excitations (CIS) level of theory. For the ground-state geometry, a conformational analysis at the Hartree-Fock level has also been carried out. We have evaluated the energy gaps between electronic levels by using Zerner's intermediate neglect of differential overlap (ZINDO) method. The species involved in the fluorescence have been experimentally characterized, and the decay-associated spectra have been obtained. PMID- 20028173 TI - Vibrational bound states of the He2Ne+ cation. AB - The vibrational bound states of the He(2)Ne(+) complex have been determined using a potential energy surface previously published by Seong et al. [J. Chem. Phys. 2004, 120, 7456]. The calculation was performed by sequential diagonalization truncation techniques in a discrete variable representation using Radau hyperspherical coordinates. There are 52 bound levels. The ground state has an energy of 605.3 cm(-1) above the absolute minimum and lies about half way to dissociation. The evaporation energy of one He atom is equal to 866.1 cm(-1). Only four levels have energies below the classical energy for dissociation, and all the other 48 states are bound by the zero-point energy of the HeNe(+) fragment. The implications of the properties of the eigenvalue spectrum and of the corresponding wave functions on the vibrational relaxation dynamics and infrared spectra of He(N)Ne(+) clusters is discussed. PMID- 20028174 TI - Single-parameter quantification of the sensitivity of a molecular collision to molecular polarization. AB - This article introduces the concept of intrinsic entropy, S, of a molecular collision. Defined in rigorously quantum mechanical terms as the von Neumann entropy of the intrinsic density matrices of reagents and products, the intrinsic entropy is a dimensionless number in the 0 < or = S < or = 1 range. Its limits are associated with situations where the collision cross section is due to a single combination of reagent and product polarizations (S = 0) or where there is absolutely no selectivity with respect to the molecular polarizations (S = 1). The usefulness of the intrinsic entropy as a quantifier of the sensitivity of a molecular collision to molecular polarizations is demonstrated with examples for the benchmark H + D(2) reaction. PMID- 20028175 TI - Quantum-mechanical study of the collision dynamics of O2(3Sigma(g)-) + O2(3Sigma(g)-) on a new ab initio potential energy surface. AB - The quantum mechanical theory for the scattering of two identical rigid rotors is reviewed and applied to the collision of O2(3Sigma(g)-) molecules using a new accurate ab initio potential energy surface (PES) for the quintet state of the composite system. The PES is based on calculations using restricted coupled cluster theory with singles, doubles, and perturbative triple excitations [RCCSD(T)] [Bartolomei; et al. J. Chem. Phys. 2008, 128, 214304.]. This PES is extended here for large intermolecular distances using the ab initio long-range coefficients of Hettema et al. [J. Chem. Phys. 1994, 100, 1297.]. Elastic and rotationally inelastic integral cross sections have been obtained by means of close coupling calculations in the subthermal energy range (center-of-mass velocities below 500 m/s). Results are compared with those obtained using a PES derived from molecular beam experiments [Aquilanti; et al. J. Am. Chem. Soc. 1999, 121, 10794.]. General agreement is found between both PESs, although the experimentally derived PES appears as somewhat more anisotropic at least for the studied energy range. There is, however, a significant difference in the absolute value of the elastic cross sections that is due to differences in the long-range dispersion interaction. The performance of the ab initio PES for higher velocities (relevant to experiments) is also explored by retaining just the isotropic component of the interaction. A satisfactory agreement is found for the shape of the glory pattern but shifted toward lower absolute values of the cross sections. PMID- 20028176 TI - Si(3P) + OH(X2Pi) interaction: long-range multipolar potentials of the eighteen spin-orbit states. AB - Eighteen spin-orbit states are generated from the open-shell open-shell Si((3)P) + OH(X(2)Pi) interacting system. We present here the behavior of the associated long-range intermolecular potentials, following a multipolar expansion of the Coulombic interaction treated up to second order of the perturbation theory, giving rise to a series of terms varying in R(-n). In the present work, we have considered the electrostatic dipole-quadrupole (n = 4) and quadrupole-quadrupole (n = 5) interactions, as well as the dipole-induced dipole-induced dispersion (n = 6) and dipole-dipole-induced induction (n = 6) contributions. The diatomic OH is kept fixed at its ground state-averaged distance, (r)(v=0) = 1.865 bohr, so that the long-range potentials are two-dimensional potential energy surfaces (PESs) that depend on the intermolecular distance R and on the bending angle gamma = angleSiGH, where G represents the mass center of OH. From the calculated properties of the monomers, such as the dipole and quadrupole moments and static and dynamic polarizabilities, we have determined and tabulated the long-range coefficients of the multipolar expansion of the potentials for each matrix elements. The isolated monomer spin-orbit splittings have been included in the final matrix, whose diagonalization gives rise to 18 adiabatic potentials. Then, the adiabatic states have been compared to potential energies given by supermolecular ab initio calculations resulting in a general good overall agreement. PMID- 20028177 TI - Nanoparticles and thin film formation in ultrashort pulsed laser deposition of vanadium oxide. AB - The ultrashort pulsed laser deposition of vanadium oxide thin films has been carried out by a frequency-doubled Nd:glass laser with a pulse duration of 250 fs. The characteristics of the plasma produced by the laser-target interaction have been studied by ICCD imaging and optical emission spectroscopy. The results confirm that an emitting plasma produced by ultrashort laser pulses is formed by both a primary and a secondary component. The secondary component consists of particles with a nanometric size, and their composition and spatial angular distribution influence the deposited films. In fact, these films, analyzed by X ray photoelectron spectroscopy, X-ray diffraction, scanning electron microscopy, and atomic force microscopy, are formed by the aggregation of a large number of nanoparticles whose composition is explained by a model based on equilibrium thermal evaporation from particles directly ejected from the target. On these basis, the presence in the films of a mixture of V(2)O(5) and VO(2) is discussed. PMID- 20028178 TI - Molecular dynamics investigation of charge carrier density influence over mobility in conjugated polymers. AB - Charge carrier mobility is known to be one of the most important efficiency delimiting factors in conducting polymer-based electronic devices. As the transport mechanism in this class of material is nonconventional, many works have tried to elucidate the charge carrier's interaction with temperature, external electric field, and the collective effects they present. Even though the multiple trap-and-release model is often used to describe these effects, its applicability is known to be restricted to electronic properties. In this work we make use of a modified version of the Su-Schrieffer-Heeger model, the most used method to describe the important properties of conducting polymer in general, to investigate the influence of temperature and carrier densities over the transport mechanisms. We obtained different regimes of temperature and carriers density influence over the systems mobility, consistent with most of the experimental data available. PMID- 20028179 TI - 355 nm multiphoton dissociation and ionization of 2, 5-dihydroxyacetophenone. AB - Multiphoton dissociation and ionization of 2,5-dihydroxyacetophenone (DHAP), an important matrix compound in UV matrix-assisted laser desorption/ionization (MALDI), is studied in a molecular beam at 355 nm using multimass ion imaging mass spectrometer and time-of-flight mass spectrometry. For laser fluence larger than 130 mJ/cm(2), nearly all of the irradiated molecules absorb at least one photon. The absorption cross section was found to be sigma = 1.3(+/-0.2) x 10( 17)cm(2). Molecules excited by two photons quickly dissociate into fragments. The major channels are (1) C(6)H(3)(OH)(2)COCH(3) --> C(6)H(3)(OH)(2)CO + CH(3) and (2) C(6)H(3)(OH)(2)COCH(3) --> C(6)H(3)(OH)(2) + COCH(3). Molecules absorbing three or more photons become parent ions or crack into smaller ionic fragments. The concentration ratio of ions (parent ions and ionic fragments) to neutral fragments is about 10(-6):1. Changing the molecular beam carrier gas from He at 250 Torr to Ar at 300 Torr results in molecular beam clustering (dimers and trimers). Multiphoton ionization of clusters by a 355 nm laser beam produces only dimer cations, (C(6)H(3)(OH)(2)COCH(3))(2)(+). Protonated clusters or negatively charged ions, observed from a solid sample of DHAP using 355 nm multiphoton ionization, were not found in the molecular beam. The experimental results indicate that the photoionization occurs in the gas phase after DHAP vaporizes from the solid phase may not play an important role in the MALDI process. PMID- 20028180 TI - Potential energy surface for the H2O-H2 system. AB - In the present paper, we introduce a representation of the potential energy surface for the H(2)O...H(2) system based on orthogonal vectors, assuming that the two molecules are rigid. We represent the interaction potential by an expansion in real hyperspherical harmonics depending on the distance between the centers of mass of the two molecules and on four angles, which account for two contributions: an external one depending on the three angle variables which define the mutual orientation of the two molecules and an internal one expressed by the angle which describes the position of the oxygen atom in H(2)O with respect to the H(2)O...H(2) system. The surface was generated in the framework of the supermolecular approach, using the counterpoise-corrected interaction energies at the MP2/aug-cc-pVQZ level. Comparisons with other recent work are presented and features of the representation discussed. PMID- 20028181 TI - The OCS S L3MM auger spectrum and angular distributions studied by photoelectron auger electron coincidence experiments. AB - The selectivity of the photoelectron-Auger electron coincidence technique has been used to isolate the S L(3)MM contribution to the L(2,3)MM Auger spectrum in OCS. In this way, a direct comparison of the energies, widths, and relative intensities of the final OCS(2+) states with theoretical calculations has been achieved. Moreover, the angular distributions of some selected Auger electrons have been measured in coincidence with the photoelectron at two different photon energies. In contrast with the results of noncoincidence measurements, the coincidence angular distributions show a significant asymmetry and a dependence on the photon energy and OCS(2+) final state. PMID- 20028182 TI - Theoretical and experimental study of valence-shell ionization spectra of guanine. AB - The full valence-shell ionization spectra of the four most stable guanine tautomers were studied theoretically. The third-order algebraic-diagrammatic construction (ADC(3)) method for the one-particle Green's function was used to calculate the energies and relative intensities of the vertical ionization transitions. For low-lying transitions, the influence of planar and nonplanar guanine configurations on the ionization energies, as well as the convergence of the results with respect to basis set was studied at the level of the outer valence Green's function (OVGF) approximation scheme. The results of the calculations were used to interpret recent synchrotron radiation valence-shell photoionization spectra of guanine in the gas phase under thermal equilibrium conditions. The photoelectron spectrum was modeled by summing individual tautomer spectra weighted by Boltzmann population ratios (BPR) of tautomers from our previous high-level ab initio thermochemical calculations. The theoretical spectra are in good agreement with the experimental results, providing assignments of most observed structures and offering insight into tautomerism of guanine in the gas phase. The first six molecular orbitals give rise to single hole states with a binding energy of about 7-12 eV. At higher binding energy the spectral features are mainly due to satellite states. PMID- 20028183 TI - Electron-wavepacket reaction dynamics in proton transfer of formamide. AB - We apply the semiclassical Ehrenfest theory, which provides electron wavepacket dynamics coupled to nuclear motion, to a study of water-assisted proton relay in formamide compared with a forced proton transfer in gas phase, both of which are associated with the tautomerization. We start with the enol (imidic acid) form HO CH=NH and track its proton transfer process to the keto (amide) form O=CH-NH(2). Identifying the fact that this is indeed a "proton transfer process" rather than hydrogen-atom migration in terms of radical character on the proton, we show a collective quantum flux of electrons, which flows backward against the proton motion. This backward flux compensates the electrons tightly covering the proton, as represented in the Mulliken charge. The enol form formamide is one of the simplest species in the group O=CR(1)-NHR(2), which is a unit of polypeptide. In the gas phase, the nitrogen atom may have a pyramidal structure as in the ammonium molecule; therefore, the C-N bond may allow low barrier rotation along it. This rotation is strongly prohibited by the formation of the double bond C horizontal lineN induced by the proton transfer. Not only the dynamical process of proton transfer itself but also the electronic structures left behind are greatly affected by the presence of water molecule(s) and polar solvents. In discussing the relative stability of the formamide after the proton transfer, the following resonance structures are frequently mentioned, O(-)-CH=N(+)H(2) <--> O=CH-NH(2). Here we address the dynamical manifestation of the resonance structures in terms of our dynamical electron theory. PMID- 20028184 TI - Analytical derivation of row-orthonormal hyperspherical harmonics for triatomic systems. AB - Hyperspherical harmonics for triatomic systems as functions of row-orthonormal hyperspherical coordinates, (also called democratic hyperspherical harmonics) are obtained explicitly in terms of Jacobi polynomials and trigonometeric functions. These harmonics are regular at the poles of the triatomic kinetic energy operator, are complete, and are not highly oscillatory. They constitute an excellent basis set for calculating the local hyperspherical surface functions in the strong interaction region of nuclear configuration space. This basis set is, in addition, numerically very efficient and should permit benchmark-quality calculations of state-to-state differential and integral cross sections for those systems. The approach used for their derivation is new and should be applicable to systems of more than three atoms. PMID- 20028185 TI - In silico pharmacophore model for tabun-inhibited acetylcholinesterase reactivators: a study of their stereoelectronic properties. AB - Organophosphorus (OP) nerve agents that inhibit acetylcholinesterase (AChE; EC 3.1.1.7) function in the nervous system, causing acute intoxication. If untreated, death can result. Inhibited AChE can be reactivated by oximes, antidotes for OP exposure. However, OP intoxication caused by the nerve agent tabun (GA) is particularly resistant to oximes, which poorly reactivate GA inhibited AChE. In an attempt to develop a rational strategy for the discovery and design of novel reactivators with lower toxicity and increased efficacy in reactivating GA-inhibited AChE, we developed the first in silico pharmacophore model for binding affinity of GA-inhibited AChE from a set of 11 oximes. Oximes were analyzed for stereoelectronic profiles and three-dimensional quantitative structure-activity relationship pharmacophores using ab initio quantum chemical and pharmacophore generation methods. Quantum chemical methods were sequentially used from semiempirical AM1 to hierarchical ab initio calculations to determine the stereoelectronic properties of nine oximes exhibiting affinity for binding to GA-inhibited AChE in vivo. The calculated stereoelectronic properties led us to develop the in silico pharmacophore model using CATALYST methodology. Specific stereoelectronic profiles including the distance between bisquarternary nitrogen atoms of the pyridinium ring in the oximes, hydrophilicity, surface area, nucleophilicity of the oxime oxygen, and location of the molecular orbitals on the isosurfaces have important roles for potencies for reactivating GA-inhibited AChE. The in silico pharmacophore model of oxime affinity for binding to GA inhibited AChE was found to require a hydrogen bond acceptor, a hydrogen bond donor at the two terminal regions, and an aromatic ring in the central region of the oximes. The model was found to be well-correlated (R = 0.9) with experimental oxime affinity for binding to GA-inhibited AChE. Additional stereoelectronic features relating activity with the location of molecular orbitals and weak electrostatic potential field over the aromatic rings were found to be consistent with the pharmacophore model. These results provided the first predictive pharmacophore model of oxime affinity for binding toward GA-inhibited AChE. The model may be useful for virtual screening of compound libraries to discover and/or custom synthesize more efficacious and less toxic reactivators that may be useful for GA intoxication. PMID- 20028186 TI - Air- and light-stable superhydrophobic colored surfaces based on supported organic nanowires. AB - In this work, we report on a new type of superhydrophobic material consisting of supported organic nanowires prepared by vacuum deposition. Different intensely colored surfaces with water contact angles as high as 180 degrees can be fabricated depending on the composition, morphology, and density of the nanowires. These surfaces are stable in air and under intense light irradiation. The wettability properties of coatings made of metalloporphyrins and metallophthalocyanines nanowires as well as other heterostructured binary and open core@shell nanowires are studied. PMID- 20028187 TI - Developing strategies to enhance health services research capacity in a predominantly rural Canadian health authority. AB - CONTEXT: This article outlines the planning, implementation and preliminary evaluation of a research capacity building (RCB) initiative within a predominantly rural Canadian health authority, Interior Health (IH), including initiative characteristics and key activities designed to initiate and enhance health services research capacity within the organization. Interior Health is one of 5 geographic health authorities in British Columbia. Over half of the population IH serves is considered to be rural/remote (approximately 3 people/km2), contributing to difficulties in sharing research information (ie geographical distance to meet in-person and a diverse set of needs and/or priority topics that warrant research support). An initial assessment of IH research capacity in 2006, using an organizational self-assessment tool and discussions with key stakeholders, revealed a need for enhanced communication of health research results, research education and networking opportunities for staff at all levels of the organization. Staff noted barriers to using and sharing research such as lack of time, resources and skills for, and value placed on, participating in research, as well as lack of awareness of linkages with local academic health researchers, including faculty located at two universities within the region. In response to this baseline assessment and stakeholder feedback, short-term funding has allowed for the initial development of RCB strategies in both urban and rural/remote areas of the region, including: IH Research Brown Bag Lunch Seminars; IH Research Skills Workshop Series; literature syntheses/summaries on priority topic areas; research collaboration/partnerships with health authorities, research networks and academic researchers; and an annual IH Research Conference. ISSUE: Although currently a poorly defined term, RCB is a concept that speaks to the need for improvement in the skills and assets that can facilitate the production and application research. It is difficult to gauge the progress of RCB initiatives when there is debate as to what the optimal outcomes and indicators of success are. Most definitions of RCB have focused on enhancing the ability to do research; however, there appears to be growing support for a more inclusive definition that also addresses the ability to use and apply research. The use and application of existing research findings, often referred to as knowledge translation and exchange (KTE), is one means of building organizational research capacity, and is particularly important within a rural health region where time, resources, and research skills are often limited. LESSONS LEARNED: Dedicated RCB resources and staff support, as well as enthusiasm, academic partnerships, and identification of research 'champions' within the organization, have been critical in building research capacity within the region. Video- and teleconferencing, as well as webcasts, have allowed for expansion of RCB activities to rural/remote communities. Preliminary evaluation parameters to date suggest that the information translated during the RCB activities is motivating different groups within IH to initiate their own research and/or KTE strategies. Although preliminary results indicate improvements in research capacity within the organization, barriers to research participation such as time, funding, and communication are still evident 3 years post-implementation. Additional challenges to building research capacity within a rural health authority include geographical distances, diverse 'hot'/priority topics in need of research support, lack of protected time and limited research related human resource capacity. The translation of research evidence and enhancement of staff research skills through the IH RCB initiatives has helped to achieve new standards of excellence in the planning, management and delivery of all health services across the predominantly rural health authority. PMID- 20028188 TI - Is there a role for erythropoietin in cardiovascular disease? AB - IMPORTANCE OF THE FIELD: Despite the advances in the cardiovascular field, cardiovascular diseases remain an important health problem with a high mortality rate. Novel therapeutic attempts that target myocardial ischemia and heart failure offer attractive adjuncts and/or alternatives to commonly employed regimens. The development of novel laboratory technologies over the last decade has led to substantial progress in bringing new therapies to the bedside. AREAS COVERED IN THIS REVIEW: Current experimental and clinical trials in the use of erythropoietin (EPO) in cardiovascular diseases are reviewed. WHAT THE READER WILL GAIN: This review will widen knowledge of the therapeutic potential of EPO's non-erythropoietic beneficial effects in a clinical cardiovascular setting. TAKE HOME MESSAGE: Results from preclinical trials regarding the non-erythropoietic effects of erythropoietin are really encouraging. Further clinical studies are warranted to define the beneficial role of EPO in the clinical setting of coronary artery disease, heart failure and peripheral artery disease. PMID- 20028189 TI - Weight status of children and adolescents in a telepsychiatry clinic. AB - The prevalence of overweight and obesity is approximately 32% among children and adolescents in the United States. Comorbid conditions associated with pediatric overweight and obesity include psychiatric conditions. The purpose of this study was to determine the prevalence of overweight and obesity among children and adolescents presenting for consultation from rural communities to the UC Davis Telemedicine Program (UCDTP), as well as to collect preliminary data to design an integrated disease management program for children and adolescents with obesity and mental illness. Patients aged 21 and under seen for psychiatric consultation at the UCDTP between 2004 and 2006 were included. Retrospective medical record review was conducted to determine the major psychiatric diagnoses, height, weight, body-mass index, and weight status (underweight/at risk for underweight, normal weight, overweight, or obese) for each patient. Of the 230 patients referred, a total of 121 patients had both height and weight values documented. Three patients were underweight; 51 were normal weight; 28 were overweight; 39 were obese. The most common psychiatric diagnoses in the 121 patients were attention deficit/hyperactivity disorder (ADHD; n = 40), bipolar disorder (n = 36), and depression (n = 31). The most common psychiatric diagnoses in patients with available weight and height data who were overweight and obese were bipolar disorder (n = 20), depression (n = 18), and ADHD (n = 17). Approximately 55% of child and adolescent patients seen for telepsychiatry consultation whose charts documented height and weight measurements were overweight or obese. Psychiatric diagnoses in overweight youngsters need to be researched further to determine whether the weight change is primary or secondary to mood and/or to treatments, such as medication. At such a high rate of comorbidity, monitoring the weight status of young psychiatric patients in this population is indicated. PMID- 20028190 TI - Online, video-based patient education improves melanoma awareness: a randomized controlled trial. AB - Despite efforts to increase melanoma awareness in the general public, research is scant regarding effective methods of education delivery. The purpose of this study was to evaluate the effectiveness of online, video-based education to increase melanoma awareness and knowledge. In this randomized controlled trial, 78 subjects received either an online, video-based education or written educational material (in the form of a pamphlet) on the description and detection of melanoma. Questionnaires were developed that assessed subjects' understanding of melanoma at baseline and at 1 month after receiving the educational intervention. Subjects in the online video group demonstrated significantly increased melanoma knowledge compared to those in the written education group, as measured by improvement from baseline on a 10-item questionnaire (2.03 and 0.72 improvement, respectively, p = 0.03). Furthermore, compared to those in the written education group, video group subjects rated their intervention significantly higher in terms of usefulness and appeal. The use of online videos may represent an effective method to raise melanoma awareness. PMID- 20028191 TI - Caring in telehealth. AB - The overall goal of this study was to explore and describe the perceptions of advanced practice nurses (APNs) about caring while providing primary care using telehealth technology. This study used naturalistic inquiry methodology to elicit the subjective perceptions and reflections of a sample of APNs about how they convey caring in the context of telehealth. Thirteen APNs, selected by purposive and snowball sampling, participated in the study. The data for the study consisted of interviews conducted by e-mail using a semistructured interview guide. Data analysis used the constant comparison method; rigor and trustworthiness of the study procedures were established using the criteria of credibility, confirmability, dependability, and transferability. The findings of this study revealed that the APNs conveyed caring to their telehealth patients by (1) being with them, (2) personifying the images, and (3) possessing certain attributes. The major constructs that emerged from the data together formed a model of how APNs conveyed caring in telehealth. The findings provide insights and increase the understanding of how caring in telehealth was perceived by APNs. The findings of the study can make important contributions in improving our profession's preparation of future telehealth APNs. The study findings also can lend themselves to developing an instrument to measure caring in telehealth. The study findings also contribute to the nursing literature in a number of ways. PMID- 20028192 TI - Ride comfort analysis with physiological parameters for an e-health train. AB - Transportation by train has numerous advantages over road transportation, especially with regard to energy efficiency, ecological features, safety, and punctuality. However, the contrast in ride comfort between standard road transportation and train travel has become a competitive issue. The ride comfort enhancement technology of tilting trains (TTX) is a particularly important issue in the development of the Korean high-speed railroad business. Ride comfort is now defined in international standards such as UIC13 and ISO2631. The Korean standards such as KSR9216 mainly address physical parameters such as vibration and noise. In the area of ride comfort, living quality parameter techniques have recently been considered in Korea, Japan, and Europe. This study introduces biological parameters, particularly variations in heart rate, as a more direct measure of comfort. Biological parameters are based on physiological responses rather than on purely external mechanical parameters. Variability of heart rate and other physiological parameters of passengers are measured in a simulation involving changes in the tilting angle of the TTX. This research is a preliminary study for the implementation of an e-health train, which would provide passengers with optimized ride comfort. The e-health train would also provide feedback on altered ride comfort situations that can improve a passenger's experience and provide a healthcare service on the train. The aim of this research was to develop a ride comfort evaluation system for the railway industry, the automobile industry, and the air industry. The degree of tilt correlated with heart rate, fatigue, and unrelieved alertness. PMID- 20028193 TI - Usefulness of remote monitoring in cardiac implantable device follow-up. AB - The conventional approach to cardiac implantable device follow-up involves patients attending regularly scheduled in-clinic visits to verify the functionality and safety of the implanted devices. Due to increasing patient volumes, as well as the time, capital, and human resources required, such routine device follow-up contributes a significant resource burden to already overstrained clinics and electrophysiology departments, as well as to patients and caregivers in terms of travel and time. Continuous device status monitoring via remote means (i.e., remote device monitoring) allows clinics the ability to immediately be aware of changes in device functionality to proactively respond to actionable events, and may represent a safe, effective, and cost-saving alternative to conventional in-clinic follow-up programs. The currently available systems to remotely transmit device data combine leading medical and information technology using Web-based networks that promptly provide physicians with data comparable to in-clinic follow-up visits and offer patients a high level of convenience. PMID- 20028194 TI - Comparison of sequential rosuvastatin doses in hypercholesterolaemia: a meta analysis of randomised controlled trials. AB - BACKGROUND: Rosuvastatin is an effective treatment for patients with hypercholesterolaemia. However, the incremental benefit and risk of increasing through the licensed dose range have not been comprehensively assessed across all available clinical trials. RESEARCH DESIGN AND METHODS: The literature databases CENTRAL, EMBASE, and MEDLINE were searched in April 2008 for trials with comparisons of sequential licensed rosuvastatin dosages: 5 vs. 10 mg/day, 10 vs. 20 mg/day, and 20 vs. 40 mg/day. Clinical trial registries were also searched. For benefit outcomes, weighted mean differences were derived using the inverse variance method. For risk outcomes, the Mantel-Haenszel method was used to calculate a summary relative risk. RESULTS: The meta-analysis included 26 trials. The results demonstrated significantly favourable changes in low-density lipoprotein cholesterol level with increasing dosage (by 6.25, 5.84, and 5.03 percentage points for 10 vs. 5 mg/day, 20 vs. 10 mg/day, and 40 vs. 20 mg/day, respectively), and also in the ratios of total cholesterol to high-density lipoprotein cholesterol (HDL-C) and apolipoprotein B to apolipoprotein A-I (all p < 0.00001). In addition, a significantly favourable change in HDL-C level was found with 20 mg/day over 10 mg/day (p = 0.02). Among the primary tolerability comparisons, no significant differences in risk were seen for muscular, hepatic, or renal adverse events, with only one exception: the risk of proteinuria by urine dipstick testing was significantly higher with rosuvastatin 40 mg/day than 20 mg/day (p = 0.01). The efficacy outcomes assessed in this meta-analysis are limited to surrogate markers of morbidity and mortality. CONCLUSIONS: This meta analysis provides evidence for improved efficacy in treating patients with hypercholesterolaemia with each sequential titration of rosuvastatin and a generally consistent tolerability profile across the dose range. PMID- 20028195 TI - Metabolism and distribution of two highly potent and selective peptidomimetic inhibitors of matriptase. AB - Matriptase is a serine protease expressed by several types of cancer cells and it participates in tumour growth and progression through the activation of hepatocyte growth factor (HGF) and urokinase-type plasminogen activator (uPA). The metabolism of two potent and selective peptidomimetic inhibitors of matriptase (CJ-1737 and CJ-672) was examined in vitro with enzyme preparations (9000g supernatants, microsomes, and plasma) from dog, pig, rat, and human. It was found that both compounds displayed interesting species-dependent differences. Though CJ-1737 was not metabolized by microsomes, by 9000g supernatants from all species, or by human or rat plasma, canine and porcine plasma enzymes rapidly hydrolysed this compound. In contrast, CJ-672 was metabolized exclusively by enzymes from human liver (microsomes and 9000g supernatants) via a two-step metabolic pathway. Additionally, the distribution of both compounds was investigated in mice. The highest amounts were measured in the kidney and liver, followed by the spleen, lung, and heart. In contrast to CJ 1737, high concentrations of CJ-672 were detected in the colon, indicating an additional biliary excretion. In summary, this work clarifies both the metabolism and distribution of two new matriptase inhibitors and demonstrates important metabolic differences between human enzymes and those from commonly used laboratory animals. PMID- 20028196 TI - Noise and baseline wandering suppression of ECG signals by morphological filter. AB - Electrocardiogram (ECG) signals describe the electrical activity of the heart, and are universally by physicists in the diagnosis of cardiac pathologies. However, during the acquisition of ECGs they are often contaminated with different sources of noise, making interpretation difficult. Different techniques have been used to filter the ECG signal, in order to optimize the signal to noise ratio (S/N). In this paper, an approach based on morphological filtering is developed in order to filter the ECG. Morphological filtering is concerned with the detection of the ECG morphology, therefore allowing the suppression of noises and particularly baseline wandering. The implemented filter is evaluated using signals taken from the MIT-BIH ECG universal database. The results show that the performance of this filter is good compared with other filtering techniques. PMID- 20028197 TI - Endometrial progesterone receptors and levonorgestrel as emergency contraceptive. PMID- 20028198 TI - Natural killer cell-mediated cytokine response among HIV-positive south Indians with pulmonary tuberculosis. AB - Natural killer (NK) cells control Mycobacterium tuberculosis infection mainly through secreted cytokines. Cytokine dysregulation among HIV may cause rapid disease progression. Our objective was to examine whether impaired production of innate cytokines are responsible for cytokine dysregulation during HIV infection. The study included 30 subjects each of normal healthy subjects (NHS), pulmonary tuberculosis patients (TB), HIV-infected individuals (HIV), and HIV-TB co infected patients (HIV-TB). Intracellular cytokine staining method was used to enumerate the cytokine-positive NK cells. Unlike NHS (100%), only 27% of HIV-TB and 57% of HIV infected patients have detectable plasma interleukin (IL)-15 levels that signify impaired rather than decreased IL-15 production. Basal type 1 cytokine (IL-2, interferon-gamma [IFN-gamma], and tumor necrosis factor-alpha [TNF-alpha])-secreting NK cells (NK1 cytokines) were decreased significantly (P < 0.05) in TB, HIV, and HIV-TB, when compared with NHS. Stimulation with M. tuberculosis H37Rv enhanced the NK1 cytokines in NHS (P < 0.05), but not in other groups. With IL-15+IL-12 stimulation, we found increased NK1 cytokines (IL-2 and IFN-gamma) in HIV (P < 0.05), but not in HIV-TB, when compared to unstimulated condition. Supplementing IL-15+IL-12 has potential in improving the frequency of NK1 cytokines for HIV, but not HIV-TB, suggesting that TB influences cytokine response during HIV infection. PMID- 20028200 TI - Molecular characterization of feline type I interferon receptor 2. AB - The cDNA sequence of feline interferon receptor 2 (feIFNAR2) was generated using RT-PCR method in present study. This gene included 1,572 bp and encoded a 523 aminoacid (aa) protein with a 35 aa signal peptide. The deduced protein shared 61% amino acid identity to the human IFNAR2. There were two fibronectin type III (FBN-III) domains of about 110 residues in the extracellular domain. Homology modeling of feIFNAR2 presented a similar structure with other IFN receptors. The ELISA and FACS experiments demonstrated that the protein could bind to feIFN alpha or feIFN-omega. However, antiviral activity assay found that feIFN-omega had broader species spectrum compared with feIFN-alpha. To define the functional differences, several point mutations of feIFNAR2 were constructed and the relative affinities of feIFN-alpha or feIFN-omega for feIFNAR2 and mutants were evaluated. The results suggested that feIFN-alpha and feIFN-omega had different binding sites on feIFNAR2. T75 and M77 on feIFNAR2 were hotspots for binding to feIFN-alpha, but not to feIFN-omega. These findings suggested that the cloned feline IFNAR2 interacted with both feIFN-alpha and feIFN-omega, however, not sharing the same binding sites. PMID- 20028199 TI - The cost effectiveness and budget impact of natalizumab for formulary inclusion. AB - BACKGROUND: Crohn's disease (CD) and multiple sclerosis (MS) are debilitating autoimmune diseases, which represent a substantial cost burden in the context of managed care. As a corollary, there is an unmet pharmacotherapeutic need in patient populations with relapsing forms of MS, in addition to populations with moderately to severely active CD with evidence of inflammation who have experienced an inadequate response to other mainstream therapies. The purpose of this study was to analyze the clinical and economic data associated with natalizumab (Tysabri) and to determine the potential impact of its formulary inclusion in a hypothetical health plan. FINDINGS: Regarding MS, the implemented cost-effectiveness and budget-impact models demonstrated an anticipated reduction in relapse rate of 67% over 2 years, and a total therapy cost of $72,120 over 2 years, equating to a cost per relapse avoided of $56,594. With respect to the model assumptions, the market share of natalizumab would experience an increase to 8.5%, resulting in a total per-member, per-month healthcare cost increase of $0.003 ($0.002 for pharmacy costs and $0.001 for medical costs). Regarding CD, over a 2-year period outlined by the model, natalizumab produced the highest average time in remission, steroid-free remission, and remission or response in comparison to the other agents. The mean total costs associated with the initiation of natalizumab, infliximab, and adalimumab were $68,372, $62,090, and $61,796, respectively. Although natalizumab's costs were higher, the mean time spent in remission while on this medication was 4.5 months, as opposed to 2.4 months for infliximab and 2.9 months with adalimumab. This shift in market share was used to estimate the change in total costs (medical + pharmacy), and the per member per-month change for the model's base case was calculated to be $0.035. LIMITATIONS: The aforementioned cost-effectiveness results for natalizumab in the treatment for CD and MS were limited by the model's predetermined assumptions. These assumptions include anticipated reduction in relapse rate after 2 years of therapy and acquisition costs in the MS model, as well as assuming a certain percentage of patients were primary and secondary failures of TNFalpha inhibitor therapy in the CD model. CONCLUSION: The evidence presented here demonstrates that natalizumab provides clinical practitioners with another tool in their fight against both MS and CD, albeit by way of a different mechanism of action. After a thorough review of the evidence, the authors find that natalizumab has been shown to be relatively cost effective in the treatment of both conditions from a payer perspective; the therapy adds a new option for those patients for whom conventional treatment was unsuccessful. PMID- 20028205 TI - Gram-positive and gram-negative bacteria induce different patterns of cytokine production in human mononuclear cells irrespective of taxonomic relatedness. AB - Upon bacterial stimulation, tissue macrophages produce a variety of cytokines that orchestrate the immune response that clears the infection. We have shown that Gram-positives induce higher levels of interleukin-12 (IL-12), interferon gamma (IFN-gamma), and tumor necrosis factor (TNF) from human peripheral blood mononuclear cells (PBMCs) than do Gram-negatives, which instead induce more of IL 6, IL-8, and IL-10. Here, we study whether these patterns follows or crosses taxonomic borders. PBMCs from blood donors were incubated with UV-inactivated bacteria representing 37 species from five phyla. IL-12, TNF, IL-1beta, IL-6, IL 8, and IL-10 were measured in the supernatants after 24 h and IFN-gamma after 5 days. Irrespective of phylogenetic position, Gram-positive bacteria induced much more IL-12 (nine times more on average) and IFN-gamma (seven times), more TNF (three times), and slightly more IL-1beta (1.5 times) than did Gram-negatives, which instead induced more IL-6 (1.5 times), IL-8 (1.9 times), and IL-10 (3.3 times) than did Gram-positives. A notable exception was the Gram-positive Listeria monocytogenes, which induced very little IL-12, IFN-gamma, and TNF. The results confirm the fundamental difference in innate immune responses to Gram positive and Gram-negative bacteria, which crosses taxonomic borders and probably reflects differences in cell wall structure. PMID- 20028206 TI - Differentiation of human SH-SY5Y neuroblastoma cells by all-trans retinoic acid activates the interleukin-18 system. AB - The clinical prognosis of children with high-stage neuroblastoma is still poor. Therapeutic approaches include surgery and cellular differentiation by retinoic acid, but also experimental interleukin-based immune modulation. However, the molecular mechanisms of all-trans retinoic acid (ATRA)-induced differentiation of neuroblastoma cells are incompletely understood. Herein, we examined the effect of ATRA on the activity of the interleukin-18 (IL-18) system in human SH-SY5Y neuroblastoma cells. It is shown that SH-SY5Y cells express IL-18 receptor (IL 18R) and the secreted antagonist IL-18-binding protein (IL-18BP), but no IL-18. SH-SY5Y cells are highly sensitive to ATRA treatment and react by cellular differentiation from a neuroblastic toward a more neuronal phenotype. This was associated with induction of IL-18 and reduction of IL-18BP expression, while IL 18R expression remained stable. Thereby, we identified the IL-18 system as a novel target of ATRA in neuroblastoma cells that might contribute to the therapeutic properties of retinoids in treatment of neuroblastoma. PMID- 20028207 TI - Biochemical monitoring of the early endocytic traffic of the type I interferon receptor. AB - The type I interferon (IFN) receptor consists of two transmembrane chains IFNAR1 and IFNAR2, associated with the tyrosine kinases Tyk2 and Jak1, respectively. Binding of IFN to this receptor complex induces activation of Jak/Stat and non Stat signaling pathways. Ligand binding also drives receptor internalization and sorting toward degradation or recycling. To gain insights into receptor trafficking and its relation to signaling, we performed subcellular organelle fractionation from IFN-stimulated Daudi cells and defined biochemically an early endosomal antigen-1 (EEA1)-positive compartment bearing the activated IFN receptor. Endosomes were thus purified by immunoaffinity isolation on anti-EEA1 antibodies-coated beads. The content of these purified endosomal fractions was analyzed by Western blot and proteomics. Shortly after IFN stimulation, robustly ubiquitinated IFNAR1 and a small amount of IFNAR2 were found in this endosomal compartment, which also contained tyrosine-phosphorylated Tyk2 and Jak1. These data strongly point to the prolonged interaction during traffic of the tyrosine kinases, still in an activated configuration, with the receptors. Among the major constituents of this EEA1-positive compartment, some proteins that have been implicated in IFN signaling were identified. Altogether, these observations suggest that trafficking of the IFN receptor through endosomes may regulate signaling pathways. PMID- 20028208 TI - Glutamine prevents total parenteral nutrition-associated changes to intraepithelial lymphocyte phenotype and function: a potential mechanism for the preservation of epithelial barrier function. AB - Total parenteral nutrition (TPN) results in a number of derangements to the intestinal epithelium, including a loss of epithelial barrier function (EBF). As TPN supplemented with glutamine has been thought to prevent this loss, this article further defined the impact of glutamine on EBF, and investigated potential mechanisms that contributed to the preservation of EBF. C57BL/6J male mice were randomized to enteral nutrition (control), TPN, or TPN supplemented with glutamine (TPN+GLN). Changes in intraepithelial lymphocyte (IEL)-derived cytokine expression were measured, and EBF was assessed with electrophysiologic methods and assessment of junctional protein expression. TPN resulted in a significant decline in EBF, and this loss of EBF was significantly prevented in the TPN+GLN group. Coincident with these changes was a loss of intraepithelial lymphocyte (IEL, mucosal lymphocyte)-derived IL-10 and increase in interferon gamma (IFN-gamma) expression, and a decline in IEL numbers in the TPN group. A prevention in the increase in IFN-gamma and decline in IL-10 expression was seen in the TPN+GLN group. To determine the mechanism responsible for these glutamine associated cytokine changes, we tested whether blockade of the IL-7 signaling pathway between epithelial cells (EC) and IEL would prevent these changes; however, blockade failed to influence IEL-derived cytokine changes. Glutamine supplemented TPN leads to a specific IEL-derived cytokine profile, which may account for the preservation of EBF; and such action may be due to a direct action of glutamine on the IEL. PMID- 20028209 TI - pH sensitive alginate-chitosan hydrogel beads for carvedilol delivery. AB - Alginate-chitosan hydrogel beads for carvedilol delivery were prepared to be used as a controlled pH-sensitive drug delivery system. The beads were characterized by scanning electron microscopy and Fourier transform infrared spectroscopy. The swelling ability and in vitro release of carvedilol in solutions of different pH value were investigated. They exhibited significant swelling rates when exposed to the slightly alkaline environment. The results suggest that the system has potential to be used as an effective pH-controlled delivery system for carvedilol. The release of carvedilol at various pH values was analyzed by a semi empirical equation. PMID- 20028210 TI - Development and evaluation of mathematical model to predict disintegration time of fast disintegrating tablets using powder characteristics. AB - The objective of the study was to develop a mathematical model for predicting the disintegration time of fast disintegrating tablets (FDTs) by estimating the powder characteristics of powder blend prior to compression. A combination of chitosan-alginate complex and glycine in the ratio of 50:50 was used for preparing FDTs. The developed mathematical model allowed water sorption time (WST), effective pore radius (R(eff.p)) and swelling Index (SI) of powder mixture as well as tablet crushing strength to be successfully correlated with disintegration time (DT) of FDTs. The predicted model showed that disintegration time of FDTs to be directly correlated with powder characteristics and inversely correlated with tablet crushing strength. Furthermore, a correlation of 0.97 was obtained when DT of FDTs was compared with SI/(WST * R(eff.p)). This correlation was not affected by inclusion of water soluble (ondansetron hydrochloride or metaclopramide hydrochloride) or water insoluble (domperidone) drugs in the powder blend or FDTs. These observations indicated the versatility of the mathematical model in predicting the disintegration time of FDTs by evaluating the selected characteristics of the powder blends without actually preparing the FDTs. PMID- 20028211 TI - Absent smooth muscle actin immunoreactivity of the small bowel muscularis propria circular layer in association with chromosome 15q11 deletion in megacystis microcolon-intestinal hypoperistalsis syndrome. AB - Megacystis-microcolon-intestinal hypoperistalsis syndrome (MMIHS; OMIM%249210) is a rare and severe form of congenital intestinal and urinary dysfunction and malformation. Histologic studies suggest that the predominant intestinal manifestation is smooth muscle myopathy. Molecular observations have linked the disease to the neuronal nicotinic acetylcholine receptor (etaAChR), namely the absence of a functional alpha3 subunit of the etaAChR in patients with MMIHS. We describe a case of MMIHS in association with a de novo deletion of the proximal long arm of chromosome 15 (15q11.2). Histologic evaluation revealed an appropriate light microscopic appearance of both the circular and longitudinal layers of the small bowel muscularis propria. Immunohistochemical staining for smooth muscle actin, however, was selectively absent in the circular layer, demonstrating isolated absence in a unique and previously undescribed pattern. These observations raise the possibility that the proximal long arm of chromosome 15 (15q11) may be of clinical significance in MMIHS. PMID- 20028212 TI - p53+/mdm2- atypical lipomatous tumor/well-differentiated liposarcoma in young children: an early expression of Li-Fraumeni syndrome. AB - The spectrum of lipomatous tumors differs in the adult and pediatric populations, with liposarcoma being rare in children. Nearly 10% of individuals with Li Fraumeni syndrome develop sarcomas in the first 2 decades of life; however, the frequency of sarcoma types and subtypes in this syndrome is unknown. Two atypical lipomatous tumors/well-differentiated liposarcomas (ALT/WDLS) were identified in the pathology files of our institution in young children from "classical" Li Fraumeni and Li-Fraumeni variant kindreds with a known germline TP53 mutation (Y220C) in one of the families. The patients were 5 and 6 years of age and the ALT/WDLSs were the first expression of the syndrome. The tumors had a high degree of cellular atypia and differed from sporadic ALT/WDLS by strong nuclear immunoreactivity for p53 and absent mdm2 expression. This is the first report of 2 ALT/WDLSs presenting in children before 10 years of age, both in association with Li-Fraumeni syndrome/variant. ALT/WDLS in a young child should raise the possibility of a cancer predisposition syndrome and, in this setting, the p53(+)/mdm2(-) immunophenotype might be characteristic. Recognition of this lesion and its association is important for early diagnosis and subsequent tumor surveillance in the proband and affected family members. PMID- 20028213 TI - Silver-Russell syndrome and Beckwith-Wiedemann syndrome phenotypes associated with 11p duplication in a single family. AB - Genomic imprinting is an epigenetic phenomenon resulting in differential expression of maternal and paternal alleles of a subset of genes. In the mouse, mutation of imprinted genes often results in contrasting phenotypes, depending on parental origin. The overgrowth-associated Beckwith-Wiedemann syndrome (BWS) and the growth restriction-associated Silver-Russell syndrome (SRS) have been linked with a variety of epigenetic and genetic defects affecting a cluster of imprinted genes at chromosome 11p15.5. Paternally derived and maternally derived 11p15.5 duplications represent infrequent findings in BWS and SRS, respectively. Here, we report a case in which a 6.5 Mb duplication of 11p15.4-pter resulted in SRS and BWS phenotypes in a child and her mother, respectively. Molecular analyses demonstrated that the duplication involved the maternal chromosome 11p15 in the child and the paternal chromosome 11p15 in the mother. This observation provides a direct demonstration that SRS and BWS represent specular images, both at the clinical and molecular levels. PMID- 20028214 TI - 2008 Annual Report of the American Association of Poison Control Centers' National Poison Data System (NPDS): 26th Annual Report. AB - BACKGROUND: This is the 26th Annual Report of the American Association of Poison Control Centers (AAPCC; http://www. aapcc.org ) National Poison Data System (NPDS). During 2008, 60 of the nation's 61 US poison centers uploaded case data automatically. The median upload time was 24 [7.2, 112] (median [25%, 75%]) minutes creating a real-time national exposure and information database and surveillance system. METHODOLOGY: We analyzed the case data tabulating specific indices from NPDS. The methodology was similar to that of previous years. Where changes were introduced, the differences are identified. Poison center cases with medical outcomes of death were evaluated by a team of 28 medical and clinical toxicologist reviewers using an ordinal scale of 1-6 to determine Relative Contribution to Fatality (RCF) from the exposure to the death. RESULTS: In 2008, 4,333,012 calls were captured by NPDS: 2,491,049 closed human exposure cases, 130,495 animal exposures, 1,703,762 information calls, 7,336 human confirmed nonexposures, and 370 animal confirmed nonexposures. The top five substances most frequently involved in all human exposures were analgesics (13.3%), cosmetics/personal care products (9.0%), household cleaning substances (8.6%), sedatives/hypnotics/antipsychotics (6.6%), and foreign bodies/toys/miscellaneous (5.2%). The top five most common exposures in children age 5 or less were cosmetics/personal care products (13.5%), analgesics (9.7%), household cleaning substances (9.7%), foreign bodies/toys/miscellaneous (7.5%), and topical preparations (6.9%). Drug identification requests comprised 66.8% of all information calls. NPDS documented 1,756 human exposures resulting in death with 1,315 human fatalities deemed related with an RCF of at least contributory (1, 2, or 3). CONCLUSIONS: Poisoning continues to be a significant cause of morbidity and mortality in the US. The near real-time, always current status of NPDS represents a national resource to collect and monitor US poisoning exposure cases and information calls. NPDS continues its mission as one of the few real-time national surveillance systems in existence, providing a model public health surveillance system for all types of exposures, public health event identification, resilience response and situational awareness tracking. PMID- 20028215 TI - The combined influence of the local environment and regional enrichment on bird species richness. AB - It is generally accepted that local species richness at a site reflects the combined influence of local and regional processes. However, most empirical studies evaluate the influence of either local environmental variables or regional enrichment but not both simultaneously. Here we demonstrate the importance of combining these processes to understand continental-scale richness patterns in breeding birds. We show that neither regional enrichment nor the local environment in isolation is sufficient to characterize observed patterns of species richness. Combining both sets of variables into a single model results in improved model fit and the removal of residual spatial autocorrelation. At short timescales, local processes are most important for determining local richness, but as the timescale of analysis increases, regional enrichment becomes increasingly important. These results emphasize the need for increased integration of multiple scales of processes into models of species richness. PMID- 20028216 TI - Male fecundity stimulation: conflict and cooperation within and between the sexes: model analyses and coevolutionary dynamics. AB - Female promiscuity forces the ejaculates of different males to compete for fertilization through sperm competition. In turn, competing ejaculates often influence female promiscuity and fitness, for example, when ejaculate products increase female fecundity. Here we develop theory examining situations where males stimulate female fecundity and compete for fertilization and female remating is influenced by male fecundity stimulation. We consider the fitness consequences that fecundity stimulation has simultaneously for males inseminating the same female and for the female herself, and we show that the way fecundity increases with male stimulation shifts the coevolutionary dynamics of female remating and male ejaculate expenditure from conflict to cooperation among all mating partners. When fecundity stimulation is weak and males "know" their mating roles, the second male to inseminate a female can exploit the fecundity stimulation of the first male, fostering intra- and intersexual conflict over female remating. However, in a diversity of species where fecundity more than doubles with remating, we show that the female and both males can gain from female remating, leading to intra- and intersexual cooperation over female remating. This coevolutionary perspective yields new insights into the connection between promiscuity, fecundity, and sperm competition and the complex interplay between sexual conflict and cooperation. PMID- 20028217 TI - A suspension induction for myocardial differentiation of rat mesenchymal stem cells on various extracellular matrix proteins. AB - The microenvironment of bone marrow-derived mesenchymal stem cells (MSCs) strictly regulates their differentiation. In this study, we have developed a new suspension induction method for myocardial differentiation of bone marrow-derived rat MSCs (rMSCs) in vitro on various extracellular matrix (ECM) proteins. Myocardial differentiation of rMSCs was induced with a conventional monolayer method and our suspension method. In our suspension induction, a cell suspension was treated with the medium in the presence of an inducer, incubated for 2 h under a suspension conditions, and moved to a monolayer culture on gelatin-coated, collagen type I-coated, fibronectin-coated, or polystyrene dishes until the total induction time was 24 h. We evaluated the myocardial differentiation by counting the number of colonies of beating cells, performing immunohistochemical staining, and measuring the expression of cardiac-specific gene mRNA using real-time quantitative polymerase chain reaction. We found that rMSCs induced with the conventional monolayer method did not differentiate efficiently, whereas beating cell colonies were found on ECM-coated dishes of suspension-induced cells, after 3 weeks of culture, especially on gelatin-coated dishes. The beating cells were positively stained with anti-troponin T-C antibody and expressed specific cardiac markers. In conclusion, these results demonstrated that the suspension induction followed by subsequent culture on gelatin ECM substrates is a promising method for differentiating rMSCs into cardiomyocytes in vitro. PMID- 20028218 TI - Effect of Nell-1 delivery on chondrocyte proliferation and cartilaginous extracellular matrix deposition. AB - Cartilage tissue engineering using chondrogenic growth factors is an attractive strategy to promote cartilage repair. Bone morphogenetic proteins have been widely studied for their application in cartilage repair. However, functional heterogeneity of bone morphogenetic proteins and unpredictable effects such as cyst formation may limit their therapeutic use. Thus, the use of alternative growth factors with greater osteochondral specificity may be advantageous for cartilage regeneration. Nel-like molecule-1 (Nell-1; Nel is a protein strongly expressed in neural tissue encoding epidermal growth factor-like domain) is a novel growth factor believed to specifically target cells committed to the osteochondral lineage. Mutation of the Nell-1 gene has been shown to disrupt normal cartilage growth and development in rodents. This study investigates the chondrogenic potential of recombinant human Nell-1 protein in a three-dimensional alginate hydrogel microenvironment containing rabbit chondrocytes. To provide controlled delivery and maximize biological efficiency, Nell-1 was incorporated in chitosan microparticles. Over 42 days of culture, chondrocyte proliferation and cluster formation was significantly enhanced by Nell-1 in a dose-dependent manner. Further, the clusters formed in the presence of Nell-1 contained more type II collagen and glycosaminoglycans than clusters formed within Nell-free control gels. These findings demonstrate the ability of Nell-1 to promote chondrocyte proliferation and deposition of cartilage-specific extracellular matrix materials. PMID- 20028219 TI - Dynamic mechanical loading enhances functional properties of tissue-engineered cartilage using mature canine chondrocytes. AB - OBJECTIVE: The concept of cartilage functional tissue engineering (FTE) has promoted the use of physiologic loading bioreactor systems to cultivate engineered tissues with load-bearing properties. Prior studies have demonstrated that culturing agarose constructs seeded with primary bovine chondrocytes from immature joints, and subjected to dynamic deformation, produced equilibrium compressive properties and proteoglycan content matching the native tissue. In the process of translating these results to an adult canine animal model, it was found that protocols previously successful with immature bovine primary chondrocytes did not produce the same successful outcome when using adult canine primary chondrocytes. The objective of this study was to assess the efficacy of a modified FTE protocol using adult canine chondrocytes seeded in agarose hydrogel and subjected to dynamic loading. METHOD: Two modes of dynamic loading were applied to constructs using custom bioreactors: unconfined axial compressive deformational loading (DL; 1 Hz, 10% deformation) or sliding contact loading (Slide; 0.5 Hz, 10% deformation). Loading for 3 h daily was initiated on day 0, 14, or 28 (DL0, DL14, DL28, and Slide14). RESULTS: Constructs with applied loading (both DL and Slide) exhibited significant increases in Young's modulus compared with free-swelling control as early as day 28 in culture (p < 0.05). However, glycosaminoglycan, collagen, and DNA content were not statistically different among the various groups. The modulus values attained for engineered constructs compare favorably with (and exceed in some cases) those of native canine knee (patella groove and condyle) cartilage. CONCLUSION: Our findings successfully demonstrate an FTE strategy incorporating clinically relevant, adult chondrocytes and gel scaffold for engineering cartilage replacement tissue. These results, using continuous growth factor supplementation, are in contrast to our previously reported studies with immature chondrocytes where the sequential application of dynamic loading after transient transforming growth factor-beta3 application was found to be a superior culture protocol. Sliding, which simulates aspects of joint articulation, has shown promise in promoting engineered tissue development and provides an alternative option for FTE of cartilage constructs to be further explored. PMID- 20028220 TI - Specific fibrinogen and thrombin concentrations promote neuronal rather than glial growth when primary neural cells are seeded within plasma-derived fibrin gels. AB - Fibrin gels are attractive scaffolds useful for neural tissue engineering applications. The objective of this work was to investigate the apoptotic activity, survival, proliferation, and differentiation of a mixed population of primary neural cells composed of neurons and multipotent precursor cells when cultured in fibrin gels prepared with varying concentrations of fibrinogen (5-25 mg/mL fibrinogen) and thrombin (1-125 U/mL thrombin). Within all fibrin gel formulations tested, the level of apoptosis on day 1 was low and cell survival was equivalent to levels in monolayer culture (67%). Proliferation in gels made from 5 to 12.5 mg/mL fibrinogen was also similar to that observed in monolayer culture, though a lower proliferative response was observed in 25 mg/mL fibrinogen formulations. Relative to monolayer culture, cholinergic and dopaminergic neuronal presence was enhanced, whereas glial cell growth was reduced in fibrin gel cultures. The extent to which levels were altered depended on fibrinogen and thrombin concentration. The findings here suggest the importance of fibrinogen and thrombin concentration in differentially regulating the growth and composition of neural cell populations and are of importance for neural tissue engineering strategies focused on the development of implantable scaffolds for treating neurodegenerative disorders. PMID- 20028221 TI - A model of indispensability of a large glial layer in cerebrovascular circulation. AB - We formulate the problem of oxygen delivery to neural tissue as a problem of association. Input to a pool of neurons in one brain area must be matched in space and time with metabolic inputs from the vascular network via the glial network. We thus have a model in which neural, glial, and vascular layers are connected bidirectionally, in that order. Connections between neuro-glial and glial-vascular stages are trained by an unsupervised learning mechanism such that input to the neural layer is sustained by the precisely patterned delivery of metabolic inputs from the vascular layer via the glial layer. Simulations show that the capacity of such a system to sustain patterns is weak when the glial layer is absent. Capacity is higher when a glial layer is present and increases with the layer size. The proposed formulation of neurovascular interactions raises many intriguing questions about the role of glial cells in cerebral circulation. PMID- 20028222 TI - Efficient learning and feature selection in high-dimensional regression. AB - We present a novel algorithm for efficient learning and feature selection in high dimensional regression problems. We arrive at this model through a modification of the standard regression model, enabling us to derive a probabilistic version of the well-known statistical regression technique of backfitting. Using the expectation-maximization algorithm, along with variational approximation methods to overcome intractability, we extend our algorithm to include automatic relevance detection of the input features. This variational Bayesian least squares (VBLS) approach retains its simplicity as a linear model, but offers a novel statistically robust black-box approach to generalized linear regression with high-dimensional inputs. It can be easily extended to nonlinear regression and classification problems. In particular, we derive the framework of sparse Bayesian learning, the relevance vector machine, with VBLS at its core, offering significant computational and robustness advantages for this class of methods. The iterative nature of VBLS makes it most suitable for real-time incremental learning, which is crucial especially in the application domain of robotics, brain-machine interfaces, and neural prosthetics, where real-time learning of models for control is needed. We evaluate our algorithm on synthetic and neurophysiological data sets, as well as on standard regression and classification benchmark data sets, comparing it with other competitive statistical approaches and demonstrating its suitability as a drop-in replacement for other generalized linear regression techniques. PMID- 20028223 TI - Dynamical mean field model of a neural-glial mass. AB - Our goal is to model the behavior of an ensemble of interacting neurons and astrocytes (the neural-glial mass). For this, a model describing N tripartite synapses is proposed. Each tripartite synapse consists of presynaptic and postsynaptic nerve terminals, as well as the synaptically associated astrocytic microdomain, and is described by a system of 13 stochastic differential equations. Then, by applying the dynamical mean field approximation (DMA) (Hasegawa, 2003a , 2003b ) the system of 13N equations is reduced to 13(13 + 2) = 195 deterministic differential equations for the means and the second-order moments of local and global variables. Simulations are carried out for studying the response of the neural-glial mass to external inputs applied to either the presynaptic terminals or the astrocytes. Three cases were considered: the astrocytes influence only the presynaptic terminal, only the postsynaptic terminal, or both the presynaptic and postsynaptic terminals. As a result, a wide range of responses varying from singles spikes to train of spikes was evoked on presynaptic and postsynaptic terminals. The experimentally observed phenomenon of spontaneous activity in astrocytes was replicated on the neural-glial mass. The model predicts that astrocytes can have a strong and activity-dependent influence on synaptic transmission. Finally, simulations show that the dynamics of astrocytes influences the synchronization ratio between neurons, predicting a peak in the synchronization for specific values of the astrocytes' parameters. PMID- 20028224 TI - Decoding Poisson spike trains by Gaussian filtering. AB - The temporal waveform of neural activity is commonly estimated by low-pass filtering spike train data through convolution with a gaussian kernel. However, the criteria for selecting the gaussian width sigma are not well understood. Given an ensemble of Poisson spike trains generated by an instantaneous firing rate function lambda(t), the problem was to recover an optimal estimate of lambda(t) by gaussian filtering. We provide equations describing the optimal value of sigma using an error minimization criterion and examine how the optimal sigma varies within a parameter space defining the statistics of inhomogeneous Poisson spike trains. The process was studied both analytically and through simulations. The rate functions lambda(t) were randomly generated, with the three parameters defining spike statistics being the mean of lambda(t), the variance of lambda(t), and the exponent alpha of the Fourier amplitude spectrum 1/f(alpha) of lambda(t). The value of sigma(opt) followed a power law as a function of the pooled mean interspike interval I, sigma(opt) = aI(b), where a was inversely related to the coefficient of variation C(V) of lambda(t), and b was inversely related to the Fourier spectrum exponent alpha. Besides applications for data analysis, optimal recovery of an analog signal waveform lambda(t) from spike trains may also be useful in understanding neural signal processing in vivo. PMID- 20028225 TI - EEG-fMRI fusion of paradigm-free activity using Kalman filtering. AB - We address here the use of EEG and fMRI, and their combination, in order to estimate the full spatiotemporal patterns of activity on the cortical surface in the absence of any particular assumptions on this activity such as stimulation times. For handling such a high-dimension inverse problem, we propose the use of (1) a global forward model of how these measures are functions of the "neural activity" of a large number of sources distributed on the cortical surface, formalized as a dynamical system, and (2) adaptive filters, as a natural solution to solve this inverse problem iteratively along the temporal dimension. This estimation framework relies on realistic physiological models, uses EEG and fMRI in a symmetric manner, and takes into account both their temporal and spatial information. We use the Kalman filter and smoother to perform such an estimation on realistic artificial data and demonstrate that the algorithm can handle the high dimensionality of these data and that it succeeds in solving this inverse problem, combining efficiently the information provided by the two modalities (this information being naturally predominantly temporal for EEG and spatial for fMRI). It performs particularly well in reconstructing a random temporally and spatially smooth activity spread over the cortex. The Kalman filter and smoother show some limitations, however, which call for the development of more specific adaptive filters. First, they do not cope well with the strong nonlinearity in the model that is necessary for an adequate description of the relation between cortical electric activities and the metabolic demand responsible for fMRI signals. Second, they fail to estimate a sparse activity (i.e., presenting sharp peaks at specific locations and times). Finally their computational cost remains high. We use schematic examples to explain these limitations and propose further developments of our method to overcome them. PMID- 20028226 TI - Adaptive design optimization: a mutual information-based approach to model discrimination in cognitive science. AB - Discriminating among competing statistical models is a pressing issue for many experimentalists in the field of cognitive science. Resolving this issue begins with designing maximally informative experiments. To this end, the problem to be solved in adaptive design optimization is identifying experimental designs under which one can infer the underlying model in the fewest possible steps. When the models under consideration are nonlinear, as is often the case in cognitive science, this problem can be impossible to solve analytically without simplifying assumptions. However, as we show in this letter, a full solution can be found numerically with the help of a Bayesian computational trick derived from the statistics literature, which recasts the problem as a probability density simulation in which the optimal design is the mode of the density. We use a utility function based on mutual information and give three intuitive interpretations of the utility function in terms of Bayesian posterior estimates. As a proof of concept, we offer a simple example application to an experiment on memory retention. PMID- 20028227 TI - Connectivity, dynamics, and memory in reservoir computing with binary and analog neurons. AB - Reservoir computing (RC) systems are powerful models for online computations on input sequences. They consist of a memoryless readout neuron that is trained on top of a randomly connected recurrent neural network. RC systems are commonly used in two flavors: with analog or binary (spiking) neurons in the recurrent circuits. Previous work indicated a fundamental difference in the behavior of these two implementations of the RC idea. The performance of an RC system built from binary neurons seems to depend strongly on the network connectivity structure. In networks of analog neurons, such clear dependency has not been observed. In this letter, we address this apparent dichotomy by investigating the influence of the network connectivity (parameterized by the neuron in-degree) on a family of network models that interpolates between analog and binary networks. Our analyses are based on a novel estimation of the Lyapunov exponent of the network dynamics with the help of branching process theory, rank measures that estimate the kernel quality and generalization capabilities of recurrent networks, and a novel mean field predictor for computational performance. These analyses reveal that the phase transition between ordered and chaotic network behavior of binary circuits qualitatively differs from the one in analog circuits, leading to differences in the integration of information over short and long timescales. This explains the decreased computational performance observed in binary circuits that are densely connected. The mean field predictor is also used to bound the memory function of recurrent circuits of binary neurons. PMID- 20028228 TI - Optimal decision making on the basis of evidence represented in spike trains. AB - Experimental data indicate that perceptual decision making involves integration of sensory evidence in certain cortical areas. Theoretical studies have proposed that the computation in neural decision circuits approximates statistically optimal decision procedures (e.g., sequential probability ratio test) that maximize the reward rate in sequential choice tasks. However, these previous studies assumed that the sensory evidence was represented by continuous values from gaussian distributions with the same variance across alternatives. In this article, we make a more realistic assumption that sensory evidence is represented in spike trains described by the Poisson processes, which naturally satisfy the mean-variance relationship observed in sensory neurons. We show that for such a representation, the neural circuits involving cortical integrators and basal ganglia can approximate the optimal decision procedures for two and multiple alternative choice tasks. PMID- 20028229 TI - A non-Markovian random walk underlies a stochastic model of spike-timing dependent plasticity. AB - A stochastic model of spike-timing-dependent plasticity (STDP) proposes that spike timing influences the probability but not the amplitude of synaptic strength change at single synapses. The classic, biphasic STDP profile emerges as a spatial average over many synapses presented with a single spike pair or as a temporal average over a single synapse presented with many spike pairs. We have previously shown that the model accounts for a variety of experimental data, including spike triplet results, and has a number of desirable theoretical properties, including being entirely self-stabilizing in all regions of parameter space. Our earlier analyses of the model have employed cumbersome spike-to-spike averaging arguments to derive results. Here, we show that the model can be reformulated as a non-Markovian random walk in synaptic strength, the step sizes being fixed as postulated. This change of perspective greatly simplifies earlier calculations by integrating out the proposed switch mechanism by which changes in strength are driven and instead concentrating on the changes in strength themselves. Moreover, this change of viewpoint is generative, facilitating further calculations that would be intractable, if not impossible, with earlier approaches. We prepare the machinery here for these later calculations but also briefly indicate how this machinery may be used by considering two particular applications. PMID- 20028230 TI - Representation sharpening can explain perceptual priming. AB - Perceiving and identifying an object is improved by prior exposure to the object. This perceptual priming phenomenon is accompanied by reduced neural activity. But whether suppression of neuronal activity with priming is responsible for the improvement in perception is unclear. To address this problem, we developed a rate-based network model of visual processing. In the model, decreased neural activity following priming was due to stimulus-specific sharpening of representations taking place in the early visual areas. Representation sharpening led to decreased interference of representations in higher visual areas that facilitated selection of one of the competing representations, thereby improving recognition. The model explained a wide range of psychophysical and physiological data observed in priming experiments, including antipriming phenomena, and predicted two functionally distinct stages of visual processing. PMID- 20028231 TI - In vivo motility evaluation of the grafted gastric wall with small intestinal submucosa. AB - Small intestinal submucosa (SIS) is a bioscaffold that has been showing encouraging results with appropriate tissue regeneration in the gastrointestinal tract. We investigated SIS-induced muscle contractility in vivo and in vitro to determine whether the SIS-grafted area would become physiologically functional in the long term. A 1 cm circular, full-thickness defect was created in a rodent stomach and was repaired using SIS. After 1 year, gastric motility in the SIS grafted area was recorded in awake animals using a telemetric force transducer system. Muscle was obtained from the same area where in vivo motility had been explored. Organ bath technique with electrical field stimulation was applied. Native stomach and grafts were investigated for morphology by histologic and immunohistochemical analyses. Gastric motility during fasting was enhanced by both food intake and a cholinergic agonist. A muscarinic receptor antagonist inhibited postprandial gastric motility. Responses in vivo were equivalent in grafted versus control rats. The in vitro study revealed that the distribution of muscle and nerves in SIS-regenerated tissue would be proportional to that in normal tissue. Smooth muscle, peripheral nerve, and gastric parietal cells were observed in the SIS-regenerated wall. SIS has the potential to promote a physical and physiological regeneration with intrinsic nerve migration. PMID- 20028233 TI - Protein E of Haemophilus influenzae is a ubiquitous highly conserved adhesin. AB - Protein E (PE) of nontypeable Haemophilus influenzae (NTHi) is involved in adhesion and activation of epithelial cells. A total of 186 clinical NTHi isolates, encapsulated H. influenzae, and culture collection strains were analyzed. PE was highly conserved in both NTHi and encapsulated H. influenzae (96.9%-100% identity without the signal peptide). PE also existed in other members of the genus Pasteurellaceae. The epithelial cell binding region (amino acids 84-108) was completely conserved. Phylogenetic analysis of the pe sequence separated Haemophilus species into 2 separate clusters. Importantly, PE was expressed in 98.4% of all NTHi (126 isolates) independently of the growth phase. PMID- 20028232 TI - Inkjet-based biopatterning of bone morphogenetic protein-2 to spatially control calvarial bone formation. AB - The purpose of this study was to demonstrate spatial control of osteoblast differentiation in vitro and bone formation in vivo using inkjet bioprinting technology and to create three-dimensional persistent bio-ink patterns of bone morphogenetic protein-2 (BMP-2) and its modifiers immobilized within microporous scaffolds. Semicircular patterns of BMP-2 were printed within circular DermaMatrix human allograft scaffold constructs. The contralateral halves of the constructs were unprinted or printed with BMP-2 modifiers, including the BMP-2 inhibitor, noggin. Printed bio-ink pattern retention was validated using fluorescent or (125)I-labeled bio-inks. Mouse C2C12 progenitor cells cultured on patterned constructs differentiated in a dose-dependent fashion toward an osteoblastic fate in register to BMP-2 patterns. The fidelity of spatial restriction of osteoblastic differentiation at the boundary between neighboring BMP-2 and noggin patterns improved in comparison with patterns without noggin. Acellular DermaMatrix constructs similarly patterned with BMP-2 and noggin were then implanted into a mouse calvarial defect model. Patterns of bone formation in vivo were comparable with patterned responses of osteoblastic differentiation in vitro. These results demonstrate that three-dimensional biopatterning of a growth factor and growth factor modifier within a construct can direct cell differentiation in vitro and tissue formation in vivo in register to printed patterns. PMID- 20028234 TI - Histoplasma capsulatum and Caenorhabditis elegans: a simple nematode model for an innate immune response to fungal infection. AB - Histoplasma capsulatum is a primary fungal pathogen of mammals responsible for histoplasmosis. During pathogenesis H. capsulatum yeast proliferate in phagosomes of macrophages. This extensive host/pathogen interaction involves a complex cascade of responses in both organisms. In the mammalian host, infection results in complex branched immunity that is initiated with an innate response and later induces an adaptive response but each response is difficult to resolve during fungal infection. Therefore, in an effort to identify less complex systems and to gain understanding of the host innate response to H. capsulatum, we constructed a mini-host survival assay. With this assay, we found ingestion of virulent Histoplasma capsulatum NAm 1 strain yeasts to be lethal to a Bristol-N2 Caenorhabditis elegans host. The virulent H. capsulatum NAm1 strain shows differential lethality under live/heat-killed infective conditions. Specifically, after ingestion of live yeast lethality is > or = 90% within 48 to 72 h, whereas worms ingesting heat-killed yeast reach equivalent mortality only after 10-14 days. On the other hand, ingestion of live H. capsulatum yeast of the nonvirulent NAm 1 (ura(-)) strain is no more lethal to the nematode than heat-killed yeast. Therefore, C. elegans provides an attractive model for further investigations of the ancient innate immune response during early host/pathogen (H. capsulatum/worm) interaction and pathogenesis. PMID- 20028235 TI - Characterization of a secreted aspartyl protease of the fungal pathogen Paracoccidioides brasiliensis. AB - Paracoccidioides brasiliensis is a thermally dimorphic fungus that causes paracoccidioidomycosis, a human systemic disease prevalent in Latin America. Proteases have been described as playing an important role in the host invasion process in many pathogenic microorganisms. Here we describe the identification and characterization of a secreted aspartyl protease (PbSAP), isolated from a cDNA library constructed with RNAs of mycelia transitioning to yeast cells. Recombinant PbSAP was produced in Escherichia coli, and the purified protein was used to develop a polyclonal antibody that was able to detect a 66 kDa protein in the P. brasiliensis proteome. PbSAP was detected in culture supernatants of P. brasiliensis and this data strongly suggest that it is a secreted molecule. The protein was located in the yeast cell wall, as determined by immunoelectron microscopy. In vitro deglycosylation assays with endoglycosidase H, and in vivo inhibition of the glycosylation by tunicamycin demonstrated N-glycosylation of the PbSAP molecule. Zymogram assays indicated the presence of aspartyl protease gelatinolytic activity in yeast cells and culture supernatant. PMID- 20028238 TI - Ratio- and predator-dependent functional forms for predators optimally foraging in patches. AB - Functional forms of predator-prey interactions are developed for predators optimally foraging on prey distributed in patches. The model uses mean free-path length theory to develop functional forms for two idealized behaviors of prey in patches. For congregating prey that maintain a fixed density, for example, fish schools, the predation rate has a ratio-dependent form, and predator interference depends only on predator density. For sessile prey, which maintain a fixed patch size, a new predator-dependent form emerges in which predator interference depends on both prey and predator densities. The Beddington-DeAngelis equation is a special case of the sessile form. The model provides behavioral and biological criteria with which to select the functional form and ranges of coefficients appropriate for a particular food web. Finally, the model illustrates that behavior is an essential factor in predator-prey dynamics. PMID- 20028239 TI - Time lags and the balance of positive and negative interactions in driving grassland community dynamics. AB - Both facilitative and competitive interactions occur simultaneously among plants, and the net balance between them can vary over time. Despite this, recent model fitting studies have found that negative interactions predominate. This suggests that more complex models may be necessary to uncover facilitation. Here we fitted models including seasonality, interannual variation, and time lags to survey data to test for patterns in positive and negative interactions among plants in a Michigan dry sand prairie. We hypothesized that interactions would be generally facilitative in this dry environment. Results indicate that most immediate (direct) interactions among dominant species are actually competitive, although interactions were more facilitative over the drier summer season. Interestingly, lagged density dependence was strong for all species in both seasons; it was positive for conspecific interactions and both positive and negative for heterospecific interactions. Observed lagged density dependence is likely due to effects from litter and/or past storage in rhizomes. Conspecific immediate and lagged interactions tended to be stronger than heterospecific interactions, suggesting that population dynamics in this community are driven mostly by conspecifics. Overall, the presence of strong lagged density dependence in this system suggests that it may be more widespread in plants than previously thought. PMID- 20028240 TI - The role of wood mass density and mechanical constraints in the economy of tree architecture. AB - By applying engineering theory, we found that in order to achieve a certain degree of stem mechanical stability, trees with low wood dry-mass density (rho(D)) need to produce thicker stems but invest less mass per unit stem length than those with high rho(D). Mechanical stability was expressed as the ability of the vertical stem to either support a plant's weight (i.e., the buckling safety factor) or resist wind forces without rupture. This contradicts the general notion that trees with low rho(D) are more prone to mechanical failure. Contrary to our results for stems, we predicted that high rho(D) can be more efficient than low rho(D) in terms of the mass needed to produce a branch of given length and resistance to rupture under its own weight. Such branches were also predicted to be more flexible. These predictions were generally in accordance with literature data for tropical tree species. This shows that differences in scaling rules associated with vertical self-loading, resistance to external forces, and the production of stable horizontal branches have important implications for the way in which different crown traits determine the balance between economy of crown design and mechanical stability. PMID- 20028242 TI - Transitions. PMID- 20028243 TI - Gerald L. Epstein, PhD: director, center for science, technology, and security policy, American Association for the Advancement of Science (AAAS). Interview by Madeline Drexler. AB - Over his entire career, Gerald Epstein has toiled at the nexus of science, technology, and security. From 2003 to 2009, he was Senior Fellow for Science and Security at the Center for Strategic and International Studies Homeland Security Program, where he worked on reducing biological weapons threats, improving national preparedness, and easing potential tensions between the scientific research and national security communities. Epstein came to CSIS from the Institute for Defense Analyses. From 1996 to 2001, he served in the White House Office of Science and Technology Policy. And from 1983 to 1989, and again from 1991 until its demise in 1995, Epstein worked at the Congressional Office of Technology Assessment, where he directed a study on the proliferation of weapons of mass destruction, alongside research on other global security topics. A recognized expert in biological risk reduction, Epstein was actually trained as a physicist, having received SB degrees in physics and electrical engineering from MIT, and a PhD in physics from the University of California at Berkeley. How, then, did he come to study the evolving threat from bioterrorism? "What compelled me about bioterrorism was that it was a stellar example of a topic that would lead to a train wreck between the scientific community and the security community unless they figured out how to work together," he said. "The distance between a laboratory and a very large consequence event is a lot shorter in biology than in any other field. I got into bioterrorism to help make sure that the security community doesn't get so scared of the science that it shuts it down, and that the science community isn't so oblivious of security concerns that it pays no attention to them." Epstein spoke on November 6, 2009, with contributing writer Madeline Drexler, author of Emerging Epidemics: The Menace of New Infections (Penguin, 2009), an updated version of an earlier volume. Drexler holds a visiting appointment at the Harvard School of Public Health and is a senior fellow at Brandeis University's Schuster Institute for Investigative Journalism. PMID- 20028244 TI - Assessing the impact of Melendez-Diaz on the investigation and prosecution of biological weapons incidents. AB - In June 2009, the U.S. Supreme Court asserted in Melendez-Diaz v Massachusetts that the admission of a laboratory analyst's certificate to validate forensic evidence against a defendant violated the defendant's Sixth Amendment Confrontation Clause rights. The Court stated that if a prosecution wished to use forensic science evidence against a defendant, the plaintiff must ensure that an actual lab analyst could testify live before the court as to the nature of the laboratory certificate being presented against the defendant, in order to uphold a defendant's constitutional right to confront the adverse witnesses against him or her. The opinion itself, as well as dicta in both the majority and the dissenting opinions, has potential implications for the success of any future prosecutions of alleged biological weapons use involving microbial forensics. Not only does the Melendez-Diaz opinion create an added burden on laboratory investigators, but the case called into question the reliability of the use of forensic science in the courtroom. Analysts and policymakers should be aware of this ruling and any potential impact the Court may have on the ability to successfully prosecute a biological weapons use event. PMID- 20028245 TI - Antibiotic resistance. AB - Antibiotic resistance poses serious challenges to health and national security, and policy changes will be required to mitigate the consequences of antibiotic resistance. Resistance can arise in disease-causing bacteria naturally, or it can be deliberately introduced to a biological weapon. In either case, life-saving drugs are rendered ineffective. Resistant bacterial infections are difficult to treat, and there are few new antibiotics in the drug development pipeline. This article describes how antibiotic resistance affects health and national security, how bacteria become antibiotic resistant, and what should be done now so antibiotics will be available to save lives in the future. PMID- 20028246 TI - Exposure to bioterrorism and mental health response among staff on Capitol Hill. AB - The October 2001 anthrax attacks heralded a new era of bioterrorism threat in the U.S. At the time, little systematic data on mental health effects were available to guide authorities' response. For this study, which was conducted 7 months after the anthrax attacks, structured diagnostic interviews were conducted with 137 Capitol Hill staff workers, including 56 who had been directly exposed to areas independently determined to have been contaminated. Postdisaster psychopathology was associated with exposure; of those with positive nasal swab tests, PTSD was diagnosed in 27% and any post-anthrax psychiatric disorder in 55%. Fewer than half of those who were prescribed antibiotics completed the entire course, and only one-fourth had flawless antibiotic adherence. Thirty percent of those not exposed believed they had been exposed; 18% of all study participants had symptoms they suspected were symptoms of anthrax infection, and most of them sought medical care. Extrapolation of raw numbers to large future disasters from proportions with incorrect belief in exposure in this limited study indicates a potential for important public health consequences, to the degree that people alter their healthcare behavior based on incorrect exposure beliefs. Incorrect belief in exposure was associated with being very upset, losing trust in health authorities, having concerns about mortality, taking antibiotics, and being male. Those who incorrectly believe they were exposed may warrant concern and potential interventions as well as those exposed. Treatment adherence and maintenance of trust for public health authorities may be areas of special concern, warranting further study to inform authorities in future disasters involving biological, chemical, and radiological agents. PMID- 20028247 TI - The four faces of microbial forensics. AB - The emerging field of microbial forensics played a major role in the investigation of the 2001 anthrax mailings and has been closely associated with the process of attribution, or identifying the perpetrator of a biological attack for purposes of criminal prosecution or military retaliation. Nevertheless, microbial forensics has other potential applications in intelligence, nonproliferation, and verification. This article describes the relevance of microbial forensics for a variety of law enforcement and national security missions, examines the obstacles to its broader use, and concludes with some policy recommendations. PMID- 20028248 TI - Trust across borders: responding to 2009 H1N1 influenza in the Middle East. AB - Disease surveillance networks in the Middle East, Southeast Asia, and Africa are models for the kind of transnational cooperation that can mount the needed flexible and coordinated response to the spread of 2009 H1N1 influenza and future pandemic threats. For example, members of the Middle East Consortium on Infectious Disease Surveillance (MECIDS), a regional disease surveillance network of public health experts and ministry of health officials from Israel, the Palestinian Authority, and Jordan, have coordinated the screening, laboratory testing, and risk communication strategies to detect and control 2009 H1N1 influenza. This coordination is made possible by trust and by well-exercised national and regional pandemic preparedness policies. The consortium illustrates the value of regional disease surveillance networks in shaping and managing cohesive policies on current and future threats. The MECIDS alliance partnership also exemplifies to other parts of the world that are experiencing conflict-like South Asia-that finding common ground is imperative to promoting health security and cooperation where it is most lacking and needed and that developing cohesive infectious disease control policies can build trust across the most difficult boundaries in the world. This article provides an overview of the history of MECIDS and similar networks and of the MECIDS response to 2009 H1N1 influenza. PMID- 20028249 TI - Source reduction in an anthrax-contaminated mail facility. AB - Following the 2001 bioterrorist attacks using letters containing Bacillus anthracis spores, a number of sites had to be decontaminated. Source reduction the process of reducing biocontamination levels prior to the main decontamination treatment-is a critical step in decontaminating buildings. Such activities include the removal of items from a facility either for off-site treatment and reuse or ultimate disposal, as well as the pretreatment of certain surfaces in the facility. In addition, waste disposal activities are initiated during the source reduction process and continued throughout the overall remediation process. This article presents the source reduction and waste disposal activities at the Department of State (DoS) Diplomatic Pouch and Mail Facility (SA-32) in Loudoun County, Virginia. At this facility, nearly all materials were removed before the fumigations with vaporized hydrogen peroxide. The materials that were removed from the site were divided into essential and nonessential items, and nonessential items were further categorized as recyclable or not recyclable. Removed materials were subjected to 5 different decontamination technologies, either before or after removal. The surfaces of large, fixed items and interior facility structures were pretreated and then fumigated. Environmental sampling was performed throughout the source reduction process for quality assurance purposes. The source reduction process lasted about 9 months and cost about $4.3 million. The overall remediation process took about 16 months and cost $8.6 million. PMID- 20028250 TI - Alphavirus antiviral drug development: scientific gap analysis and prospective research areas. AB - The New World alphaviruses Venezuelan equine encephalitis virus (VEEV), eastern equine encephalitis virus (EEEV), and western equine encephalitis virus (WEEV) pose a significant threat to human health as the etiological agents of serious viral encephalitis through natural infection as well as through their potential use as a biological weapon. At present, there is no FDA-approved medical treatment for infection with these viruses. The Defense Threat Reduction Agency, Joint Science and Technology Office for Chemical and Biological Defense (DTRA/JSTO), is currently funding research aimed at developing antiviral drugs and vaccines against VEEV, EEEV, and WEEV. A review of antiviral drug discovery efforts for these viruses revealed significant gaps in the data, assays, and models required for successful drug development. This review provides a description of these gaps and highlights specific critical research areas for the development of a target-based drug discovery program for the VEEV, EEEV, and WEEV nonstructural proteins. These efforts will increase the probability of the successful development of a pharmaceutical intervention against these viral threat agents. PMID- 20028251 TI - Influenza preparedness in Nebraska assisted living facilities. AB - We conducted an electronic survey to determine the state of planning for pandemic influenza in assisted living facilities. A survey instrument was designed with input from a number of experts and sent out to 275 Nebraska assisted living facilities; responses were received from 137. The survey covered approximately 20 areas related to preparedness. About one-third of assisted living facilities had a pandemic influenza plan; 45% had started stockpiling, and 55% expected significant staff shortages in a pandemic. Only 5% were willing to discharge current patients to make room for overflow hospital patients. Assisted living facilities have started pandemic influenza planning, but additional work needs to be done. These facilities are unlikely to be able to assist with hospital patient overflow in a pandemic. PMID- 20028252 TI - Prevention of biothreats: a look ahead. PMID- 20028253 TI - A new model of bioterrorism risk assessment. AB - There has been an abundance of different bioterrorist attack scenarios and consequently an unclear biodefense strategy so far. We present a framework for bioterrorism risk assessment that we believe would be useful for policymakers and understandable without needing to be an expert in this field. We retrieved the Medline database via PubMed (from January 1987 to January 2009) and cross referenced and reviewed the terms biological weapons, biological attacks, bioterror, bio(defense), bio(strategy) and epidemiologic models, and risk assessment. Additionally, we conducted an internet search with the same terms and strategy. We divided bioterrorist attacks into 3 categories: strategical (large scale), operational (middle-scale), and tactical (small-scale). A bioterrorist attack is presented as a 4-component chain model, including perpetrators, agents, means of delivery, and targets. For any of these 4 components, we propose quantitative and qualitative risk assessment parameters. Here we present a simple scoring system within our model applied to the 2001 U.S. anthrax attacks. PMID- 20028254 TI - Procalcitonin measurement to reduce antibiotic use in influenza. PMID- 20028256 TI - Effect of para-aminobenzoic acid on the course of retinal degeneration in the rd10 mouse. AB - PURPOSE: Recent evidence suggests that oxidative injury plays a significant role in the pathogenesis of retinal degenerative diseases. Para-aminobenzoic acid (PABA) is a cyclic amino acid, which may act to decrease lipid peroxidation and oxidative injury. Our aim was to evaluate the efficacy of PABA in attenuating oxidative injury and rate of retinal degeneration in the rd10 mouse. METHODS: PABA (50 mg/kg) was administered intraperitoneally six times per week in 28 rd10 mice from postnatal day 3. Twenty-four littermate control mice were similarly injected with saline. At 3, 4.5, and 6 weeks of age, electrophysiological (full field electroretinogram-ERG), quantitative histological, and immunohistochemical techniques were used to assess the course and extent of retinal degeneration. Degree of lipid peroxidation was determined by the measurement of thiobarbituric acid reactive species (TBARS) and retinal carbonyl content was quantified using the 2,4-dinitrophenylhydrazine method. RESULTS: Dark adapted mixed rod-cone ERG responses at 3 weeks of age were higher in the PABA-treated group as compared to saline control (P < 0.05). By 4.5 weeks, this protective effect was largely abolished and by 6 weeks ERG was unrecordable in both groups. However, at both 3 and 4.5 weeks of age, light-adapted cone ERG amplitudes were better preserved in PABA-treated animals. At 4.5 weeks, thickness of the outer nuclear layer was 28.6% higher in the peripheral retina of PABA-treated mice as compared to controls (P < 0.05). Quantitative immunohistochemistry revealed 2.4-fold higher red/green cone opsin content in the retinas of PABA-treated mice (P < 0.005). At both 3 and 4.5 weeks, levels of TBARS and protein carbonyls were 49%-69% lower in PABA-treated retinas (P < 0.05-0.0005), suggesting less oxidative injury. CONCLUSIONS: PABA treatment may protect retinal function and attenuate the course of retinal degeneration in rd10 mice. Biochemical parameters indicate a lower degree of oxidative injury in PABA-treated retinas. PABA may potentially serve as an addition to antioxidative treatment for retinal and macular degenerations. PMID- 20028258 TI - Dynamic alteration of low-density lipoprotein receptor after exposure to transforming growth factor-beta2 in human Tenon's capsule fibroblasts. AB - PURPOSE: The present study investigated dynamic alteration of low-density lipoprotein receptor and its binding and uptake of low-density lipoprotein (LDL) after exposure to transforming growth factor-beta(2) (TGF-beta(2)) in human Tenon's capsule fibroblasts. METHODS: Tenon's capsule fibroblasts obtained from elective cataract surgery patients were cultured and stimulated with different concentrations (0.1-10 ng/mL) of TGF-beta(2) for 24, 48, and 72 h. The LDLr mRNA and protein levels were analyzed by relative quantification real-time RT-PCR and Western blot analysis, respectively. The binding and uptake of DiO (3,3' dioctadecyloxacarbocyanine)-labeled LDL was assessed by confocal microscopy. RESULTS: Real-time RT-PCR and Western blot analyses showed similar results revealing that after exposure to TGF-beta(2), the expression of protein and mRNA of LDLr occurred in a concentration-dependent and time-dependent manner with a peak at a concentration of 1.0 ng/mL at 72 h in Tenon's capsule fibroblasts. Confocal microscopy showed that DiO-LDL binding and uptake were time-dependent, reaching saturation at approximately 6 h. CONCLUSIONS: This study shows that LDLrs were overexpressed in the activated Tenon's capsule fibroblasts in a concentration-dependent and time-dependent manner after exposure to TGF-beta(2). The results suggest that LDLr in the activated Tenon's capsule fibroblasts may become a novel focus as a target receptor for controlled drug delivery, particularly in anti-scarring therapy during excessive conjunctival wound healing. PMID- 20028257 TI - Interaction of ocular hypotensive agents (PGF2 alpha analogs-bimatoprost, latanoprost, and travoprost) with MDR efflux pumps on the rabbit cornea. AB - PURPOSE: The objectives of this work were (i) to screen ocular hypotensive prostaglandin (PGF2 alpha) analogs--bimatoprost, latanoprost, and travoprost as well as their free acid forms--for interaction with efflux pumps on the cornea and (ii) to assess the modulation of efflux upon co-administration of these prostaglandin analogs. METHODS: Cultured rabbit primary corneal epithelial cells (rPCEC) were employed as an in vitro model for rabbit cornea. Transporter specific interaction studies were carried out using Madin-Darby canine kidney (MDCK) cells overexpressing MDR1, MRP1, MRP2, MRP5, and BCRP. Freshly excised rabbit cornea was used as an ex vivo model to determine transcorneal permeability. RESULTS: Cellular accumulation studies clearly showed that all prostaglandin analogs and their free acid forms are substrates of MRP1, MRP2, and MRP5. Bimatoprost was the only prostaglandin analog in this study to interact with P-gp. In addition, none of these molecules showed any affinity for BCRP. K (i) values of these prostaglandin analogs obtained from dose-dependent inhibition of erythromycin efflux in rPCEC showed bimatoprost (82.54 microM) and travoprost (94.77 microM) to have similar but higher affinity to efflux pumps than latanoprost (163.20 microM). Ex vivo studies showed that the permeation of these molecules across cornea was significantly elevated in the presence of specific efflux modulators. Finally, both in vitro and ex vivo experiments demonstrated that the efflux of these prostaglandin analogs could be modulated by co administering them together. CONCLUSION: Bimatoprost, latanoprost, travoprost, and their free acid forms are substrates of multiple drug efflux pumps on the cornea. Co-administration of these molecules together is a viable strategy to overcome efflux, which could simultaneously elicit a synergistic pharmacological effect, since these molecules have been shown to activate different receptor population for the reduction of intraocular pressure (IOP). PMID- 20028259 TI - Evaluation of differential toxicity of varying doses of bevacizumab on retinal ganglion cells, retinal pigment epithelial cells, and vascular endothelial growth factor-enriched choroidal endothelial cells. AB - PURPOSE: To evaluate in vitro the effects of bevacizumab, an anti-vascular endothelial growth factor (VEGF) antibody, on retinal pigment epithelial cells (RPE) and retinal ganglion cells (RGC), at doses that were inhibitory to VEGF enriched choroidal endothelial cells (CEC). METHODS: Monkey CEC (RF6A), human RPE cells (ARPE-19), and rat RGC (RGC-5) were exposed for 24 h to increasing doses of bevacizumab. Cell numbers were quantified with WST-1 assay. Cell death was assessed using propidium iodide (PI) staining via flow cytometry and fluorescent microscopy. RESULTS: Bevacizumab was inhibitory to RF6A at 2.0 mg/mL (P < 0.005). No effect on cell viability was noted on ARPE-19 and RGC-5 cell lines at this particular dose of bevacizumab. These results were supported by fluorescent microscopy of PI-stained cells. CONCLUSIONS: VEGF-stimulated proliferation of CEC was inhibited by bevacizumab. Bevacizumab was not cytotoxic to human RPE and rat RGC in vitro at a dose that is inhibitory to monkey CEC. PMID- 20028260 TI - Serial retinal nerve fiber layer changes in patients with toxic optic neuropathy associated with antituberculosis pharmacotherapy. AB - PURPOSE: To analyze serial changes in retinal nerve fiber layer (RNFL) thickness after development of visual loss in patients with toxic optic neuropathy (TON) associated with antituberculosis pharmacotherapy, as well as to determine if RNFL changes correlate with clinical findings. METHODS: Five patients with TON underwent serial optical coherence tomography (OCT) and ophthalmologic examination, which consisted of visual acuity, color vision, pupillary light reflexes, visual fields, and funduscopy. RESULTS: Four of the five patients showed normal RNFL thickness on initial OCT. RNFL thickness remained within normal range throughout the follow-up period (range 5.5-50.5 months, average 22.0 +/- 24.8 months after initial visual symptoms) in 3 patients and gradually decreased in 2 patients (4 and 26.5 months after initial visual symptoms), especially in the temporal segment. All patients exhibited improved visual acuity and visual fields. Four patients showed improved color vision. CONCLUSIONS: RNFL changes may not be obvious at initial presentation, but became manifest in 2 of the 5 patients over time. OCT may not be useful in identifying the RNFL defect in the early stage, but may be helpful for the follow up of the patients with TON. PMID- 20028261 TI - Fixed-combination brimonidine/timolol as adjunctive therapy to a prostaglandin analog: a 3-month, open-label, replacement study in glaucoma patients. AB - PURPOSE: To evaluate intraocular pressure (IOP) and ocular allergy after replacement of 1 of 2 adjunctive therapy regimens (fixed-combination dorzolamide/timolol or dorzolamide/timolol plus brimonidine) with fixed combination brimonidine/timolol in glaucoma patients treated with ongoing prostaglandin analog (PGA) therapy. METHODS: This prospective, nonrandomized, open-label study involved patients on dorzolamide 2%/timolol 0.5% and a PGA who needed lower IOP and patients on brimonidine, dorzolamide 2%/timolol 0.5%, and a PGA who wanted to simplify their treatment regimens. After the baseline evaluation, patients were continued on the PGA and their other IOP-lowering medications were replaced with brimonidine 0.2%/timolol 0.5%. IOP was measured at baseline and months 1 and 3. RESULTS: In patients who replaced dorzolamide/timolol with brimonidine/timolol (n = 45), the mean (SD) IOP was 15.9 (1.4) mm Hg at baseline, 13.3 (0.9) mm Hg after 1 month (P < 0.001 vs. baseline), and 13.3 (1.0) mm Hg after 3 months (P < 0.001 vs. baseline). In patients who replaced both brimonidine and dorzolamide/timolol with brimonidine/timolol (n = 15), the mean (SD) IOP was 15.9 (5.2) mm Hg at baseline, 13.8 (1.8) mm Hg after 1 month (P = 0.053 vs. baseline), and 13.8 (1.4) mm Hg after 3 months (P = 0.079 vs. baseline). Allergy was reported in 5 patients previously treated with dorzolamide/timolol and 1 patient previously treated with brimonidine plus dorzolamide/timolol. CONCLUSIONS: For patients on multiple-drug therapy including a PGA, replacement of dorzolamide/timolol with brimonidine/timolol may help achieve a lower IOP, while replacement of brimonidine plus dorzolamide/timolol with brimonidine/timolol may help achieve as low an IOP with fewer medications. PMID- 20028262 TI - Intravitreal ranibizumab in the treatment of cystoid macular edema associated with retinitis pigmentosa. AB - PURPOSE: To report and evaluate the anatomic, clinical, and visual acuity response after intravitreal ranibizumab (IVR) injection in patients with cystoid macular edema (CME) due to retinitis pigmentosa (RP). METHODS: This study included 30 eyes of 30 patients with RP who had persistent CME at least 6 months despite medication with acetazolamide. Fifteen eyes of 15 eligible patients were treated with 0.5 mg IVR injection (treatment group). Fifteen eyes of 15 patients with the same characteristics who refused treatment were accepted as control group. The primary outcome of the study (morphologic restitution) was the complete or significant resolution of cystoid space on optic coherence tomography (OCT) without relapse or complication at 6 months. The serial changes in best corrected visual acuity (BCVA) and central foveal thickness (CFT) were measured. RESULTS: Thirteen eyes (86.6%) in the treatment group had significant resolution of CME at 6 months after single IVR injection. The difference between the 2 groups in BCVA was not statistically significant (P > 0.05). The baseline mean +/ SD CFT for the treatment and control groups were 478 +/- 88 microm and 469 +/- 75 microm, respectively (P > 0.05). At 6 months after treatment, the mean +/- SD CFT of the treatment group improved to 272 +/- 65 microm whereas that in the control group was 480 +/- 92 microm (P < 0.001). CONCLUSIONS: This investigation indicated that IVR may provide a new therapeutic approach for the treatment of CME secondary to RP. No adverse event was found to be associated with the treatment. Continued experience with IVR in this population will help establish its longer-term efficacy. PMID- 20028263 TI - Comparative study of topical application of timolol and verapamil in patients with glaucoma within 6 months. AB - INTRODUCTION: As glaucoma is one of the most significant causes of blindness, and administration of calcium channel blockers is effective in reducing intraocular pressure (IOP) in rabbits and patients with normotensive glaucoma, we administered topical verapamil 0.25% in the human eye to compare its effect with timolol 0.5% in reducing IOP. PURPOSE: To compare the effect of timolol 0.5% and topical verapamil 0.25% in patients with open-angle glaucoma. METHODS: It was a double-blinded study in which 118 eyes (59 individuals) were chosen and divided into 2 groups (30 individuals related to timolol and 29 individuals related to verapamil). Patients who used drugs (systemic or topical) that could alter IOP and those with IOP <22 mmHg were excluded from the study (19 eyes). We chose patients who did not use any drugs 24 h prior to the study. Then applanation tonometry was done exactly before the administration of drugs and 90 min later and the results were compared. RESULTS: In timolol group, mean intraocular pressure in 52 eyes (27 right eyes and 25 left eyes) decreased from 32.545 to 30.230 and mean pressure in verapamil group decreased from 33.195 to 30.835. CONCLUSION: It seems that topical verapamil has a similar effect to timolol in patients with open-angle glaucoma, so it can be considered as a lowering intraocular pressure agent in glaucoma patients. PMID- 20028264 TI - Hyperemia-associated costs of medication changes in glaucoma patients treated initially with prostaglandin analogs. AB - AIMS: To develop a model to estimate and compare the cost of changing therapy due to hyperemia in glaucoma patients treated initially either with latanoprost, bimatoprost, or travoprost monotherapy. METHODS: Data collected from the HealthCore Integrated Research Database, as part of the Glaucoma Adherence and Persistency Study (GAPS), were used to populate the model. Patients with a documented diagnosis of glaucoma who were newly treated (no ocular hypotensive medication and no glaucoma-related procedure during 6 months before first prescription) with latanoprost, bimatoprost, or travoprost monotherapy were identified. The time horizon for the base-case model was the duration of chart abstraction (mean = 4.1 years); a 3-month model also was developed. Physician reported rates of hyperemia were obtained from chart reviews of 300 patients. Transition rates reflected events related to reports of hyperemia where a physician-driven change (switch or discontinuation) in therapy was documented. The per-patient direct cost (2008) due to hyperemia-driven change in therapy was calculated as the sum of the cost of the initial prescription plus the cost of the office visit where the patient was evaluated and the decision to change therapy was made. Costs were stratified by whether patients were hyperemia free or discontinued the initial therapy due to hyperemia. RESULTS: From the sample of 13,977 newly treated patients, 8,743 patients were started on a prostaglandin monotherapy only. Of these, 5,726 received latanoprost, 1,633 were treated with bimatoprost, and 1,384 received travoprost index monotherapy. Across all treatment groups, costs among hyperemia-free patients were US$73.67 versus US$140.02 for those who discontinued the initial prostaglandin due to hyperemia. Per-patient costs were lowest in the group treated initially with latanoprost. For the base-case model, with latanoprost as the reference, total per-patient incremental costs due to hyperemia-driven change in therapy were US$5.92 for bimatoprost and US$5.43 for travoprost. Results were not highly sensitive to increases either in the incidence of hyperemia among latanoprost-treated patients or in the cost of latanoprost. CONCLUSIONS: Hyperemia results in increased overall costs in patients treated with latanoprost, bimatoprost, and travoprost. Treatment with latanoprost is associated with lower hyperemia-related costs than treatment with bimatoprost or travoprost. PMID- 20028265 TI - In vivo and in vitro investigations of fungal keratitis caused by Colletotrichum gloeosporioides. AB - PURPOSE: To report a case of fungal keratitis caused by Colletotrichum gloeosporioides, which is a rare pathogen in humans. METHODS: An 80-year-old woman developed fungal keratitis after having sustained a traumatic injury during field work. The patient was initially examined by slit-lamp biomicroscopy and the Heidelberg Retina Tomograph II-Rostock Cornea Module (HRT II-RCM). Corneal scrapings were collected and submitted for laboratory investigations. RESULTS: Many septate, hyphae-like interlocking and branching white lines were observed in the area of the infiltrate by HRT II-RCM. A tentative diagnosis of fungal keratitis was made, and the patient was treated with systemic and topical voriconazole and pimaricin ophthalmic ointment. The infectious focus resolved within 2 weeks, and there were no signs of a recurrence after 3 months of treatment with the antifungal agents. The culture of the corneal scraping grew C. gloeosporioides. CONCLUSIONS: HRT II-RCM was useful in detecting filamentous fungi in the cornea. The treatment with voriconazole and pimaricin was effective in the treatment of C. gloeosporioides keratitis. PMID- 20028266 TI - Bilateral reversible corneal edema associated with amantadine use. AB - PURPOSE: In this article, we report a case of bilateral severe reversible corneal edema caused by amantadine therapy. CASE: A 39-year-old women was referred to us for evaluation of bilateral corneal edema. Her past medical history was significant for multiple sclerosis, anorexia, and seizures. She developed painless progressive bilateral loss of vision for the past 6 months. She was evaluated by several ophthalmologists elsewhere who felt that the patient's visual loss was secondary to a nutritional deficiency as opposed to related to multiple sclerosis. She was started on vitamin B-12 medication without improvement in her symptoms. She was then evaluated by neuro-ophthalmology. The examination revealed severe bilateral corneal edema and was referred to our corneal service for further evaluation of her corneal condition. Our examination revealed best corrected visual acuity of 20/400 bilaterally. Corneal thickness was 940 microm in the right eye and 802 microm in the left. Color vision was intact. Conjunctivas were white bilaterally. Cornea evaluation revealed diffuse stromal edema and Descemet's folds and microcystic subepithelial edema with to guttae noted. Anterior chambers were deep and quiet. A specular microscopy revealed significant pleomorphism and polymegathism with an endothelial cell count of 1,504 cells in the right eye and 1,596 in the left eye. RESULTS: Review of the patient's medical information revealed therapy with amantadine 2 months prior to the appearance of the patient's symptoms as a means to control the patient's tremors. The patient experienced rapid resolution of the corneal edema within the next 2 months after discontinuation of the agent with recovery of best corrected visual acuity of 20/40 in the right eye and 20/30 in the left. CONCLUSIONS: In cases of unexplained corneal edema and in the absence of any identifiable ocular cause, a review of toxic effects of systemic medications should be performed. Early diagnosis may prevent irreversible endothelial damage. Amantadine can cause endothelial failure and needs to be considered as part of the differential diagnosis of corneal edema. PMID- 20028267 TI - Re: Association between moxifloxacin ophthalmic solution and fungal infection in patients with corneal ulcers and microbial keratitis. PMID- 20028269 TI - Role of the bile salt export pump, BSEP, in acquired forms of cholestasis. AB - Generation of bile is a key function of the liver. Its impairment leads to accumulation of cytotoxic bile salts in hepatocytes and, consequently, to liver disease. The bile salt export pump, BSEP, is critically involved in the secretion of bile salts into bile. Its function can be disturbed or abolished by inherited mutations. This will lead to progressive intrahepatic cholestais and severe liver disease. In addition to mutations, BSEP can be inhibited by acquired factors, such as xenobiotics or drugs, aberrant bile salt metabolites, or pregnancy. This inhibition will lead to acquired cholestasis. Some drugs are now known to be competitive inhibitors of Bsep. In addition, a polymorphism in the gene coding for BSEP has been identified as a potential susceptibility factor for acquired cholestasis. PMID- 20028270 TI - Potential biomarkers of muscle injury after eccentric exercise. AB - Proteomics was utilized to identify novel potential plasma biomarkers of exercise induced muscle injury. Muscle injury was induced in nine human volunteers by eccentric upper extremity exercise. Liquid chromatography-mass spectrometry identified 30 peptides derived from nine proteins which showed significant change in abundance post-exercise. Four of these proteins, haemoglobin alpha chain, haemoglobin beta chain, alpha1-antichymotrypsin (ACT) and plasma C-1 protease inhibitor (C1 Inh), met the criterion for inclusion based on changes in at least two distinct peptides. ACT and C1 Inh peptides peaked earlier post-exercise than creatine kinase, and thus appear to provide new information on muscle response to injury. PMID- 20028271 TI - Health reform and the elephant in the room. PMID- 20028272 TI - Let's drink (and eat) to our obese economic heroes. PMID- 20028274 TI - Climate change and human health: recognising the really inconvenient truth. PMID- 20028275 TI - Trouble in paradise--Tropical, Emergency and Disaster Medicine Conference and Tropical Medicine Summit, Broome, Western Australia, 22-24 May 2009. PMID- 20028276 TI - Frequent users of the Royal Flying Doctor Service primary clinic and aeromedical services in remote New South Wales: a quality study. AB - OBJECTIVE: To examine activity patterns of the Royal Flying Doctor Service of Australia (RFDS) in far western New South Wales and to determine whether frequent use of RFDS services, particularly emergency evacuations, is a useful indicator of patients who may benefit from care planning and review. DESIGN, SETTING AND PARTICIPANTS: We conducted a retrospective audit of the RFDS South Eastern Section's Broken Hill patient database. Patients with a residential address in the study area who had accessed at least one RFDS medical service between 1 July 2000 and 30 June 2005 were included in the study. MAIN OUTCOME MEASURES: Number of evacuations, clinic consultations and remote consultations; clinic usage by frequent evacuees; number of primary diagnoses recorded for frequent evacuees; number of frequent users who might benefit from multidisciplinary care or specialist shared care. RESULTS: Between July 2000 and June 2005, the number of residents requiring evacuation or remote consultations declined by 26% and 19%, respectively, and the number of residents accessing clinics declined by 6%. (Over the same period, the population of the study area fell by about 24%.) Of the 78 patients who were identified as frequent users of the evacuation service (> or = 3 evacuations/year), 34 had three or more primary diagnoses recorded; 15 were infrequent or non-users of the clinics (< or = 3 attendances/year); 53 may have benefited from multidisciplinary care, and 41 from specialist shared care. CONCLUSIONS: Simple, practical clinical review systems can help health care organisations in rural and remote communities to achieve better outcomes by identifying patients who may benefit from planned care. PMID- 20028277 TI - Geriatric ward rounds by video conference: a solution for rural hospitals. AB - OBJECTIVE: To evaluate the acceptance and cost of a ward-based geriatric consultation service delivered via a mobile videoconferencing system. DESIGN AND SETTING: Prospective observational study conducted in the geriatric unit of Toowoomba Base Hospital, Queensland, comparing a specialist consultation service delivered by videoconference (VC) with a "traditional" in-person service. The VC system was established in January 2007 and evaluated over an 18-month period. Patient satisfaction with the service was assessed by questionnaire during a 1 week period in September 2008. MAIN OUTCOME MEASURES: Hospital acceptance of the service; patient satisfaction with the service; comparative cost of providing in person and VC-mediated consultations. RESULTS: Uptake of the service increased progressively throughout the study period. Patient acceptance levels were high. The cost of video consultations for a 12-patient ward round and case conference was less than the cost of in-person consultations if the total road distance travelled by the specialist (Brisbane to Toowoomba and back) was 125 km or longer. CONCLUSION: Consultations via VC are an acceptable alternative to in person consultations, and are less expensive than in-person consultations for even modest distances travelled by the clinician. PMID- 20028278 TI - The first medical jet aircraft for the Royal Flying Doctor Service. PMID- 20028279 TI - The Drake Shake and a more bilious shade of green: a tale of mistaken latitude, altitude and tablets. PMID- 20028280 TI - The hidden trauma of organ donation. PMID- 20028281 TI - Not that day ... PMID- 20028282 TI - Organ donation: a matter of trust. PMID- 20028283 TI - "He's in the garage"--taking time at the end of life. PMID- 20028284 TI - Young, pregnant and dying--how can we provide the "right" care? PMID- 20028285 TI - As mass media evolves into "masses of media", what are the implications for our health? PMID- 20028286 TI - The content and structure of Australian television reportage on health and medicine, 2005-2009: parameters to guide health workers. AB - OBJECTIVE: To describe the content and structure of health and medical news and current affairs reportage on free-to-air television in Sydney, New South Wales. DESIGN AND SETTING: Review of content of all health-related evening news and current affairs items recorded over 47 months (May 2005-March 2009). MAIN OUTCOME MEASURES: Number and length of health-related items on news and current affairs programs, and topics covered in these (21 broad content areas and the leading 50/237 specific content areas); use of news actors, soundbite duration and apparent news triggers. RESULTS: 11,393 news items and 2309 current affairs items were analysed. Health news items lasted a median of 97 seconds. In a randomly selected sample of 251 items, items featured a mean of 2.2 news actors (3.9 in longer current affairs items). Median soundbite duration was 7.2 seconds for news items and 8.9 seconds for current affairs items. People affected by disease or injury were the most commonly featured news actors (84% of items), followed by experts and health professionals (56%). Many items (42%) appeared to be triggered by incidents, but a further 42% could have been triggered by press releases and other forms of publicity. CONCLUSIONS: Health workers wishing to participate in news coverage should be aware that complex issues are reduced to fit the time constraints and presentational formulae of the news media. Advocates should plan their communication strategies to accommodate these constraints. PMID- 20028287 TI - Content analysis of disease awareness advertisements in popular Australian women's magazines. AB - OBJECTIVE: To examine the nature of disease awareness advertising (DAA). DESIGN: Therapeutic advertisements in six popular Australian women's magazines were monitored between April 2006 and March 2007. A subset of advertisements was included in the study based on criteria derived from a definition of DAA. Unique advertisements were analysed by four independent coders. MAIN OUTCOME MEASURES: Types of advertisements and their sponsors, the types of disease information present, and the persuasive techniques utilised. RESULTS: Of 711 advertisements identified, 60 met the inclusion criteria for DAA, and 30 of these were unique. Over one-third of the advertisements were classified as "unbranded product advertisements", which promote a product without referring directly to a brand. DAA sponsored by pharmaceutical companies most often provided treatment and prevalence information. Most (22/30) advertisements used emotional appeals; 15 of these used "happiness/healthiness/wellbeing". CONCLUSIONS: The educational value of industry-sponsored DAA could be improved if regulations and guidelines stipulated disease information requirements, such as inclusion of risk-factor and symptom information. Regulators should provide guidelines for "unbranded product advertisements" and the acceptability of other persuasive techniques. Further research into DAA is required and should consider advertisements in a range of media, and behavioural responses. PMID- 20028289 TI - A lifetime pursuit of diabetes through chance. PMID- 20028290 TI - "Not in your lifetime, Ian!". PMID- 20028292 TI - Research to improve health practice and policy. PMID- 20028294 TI - Rational allocation of Australia's research dollars: does the distribution of NHMRC funding by National Health Priority Area reflect actual disease burden? AB - OBJECTIVES: To explore National Health and Medical Research Council (NHMRC) funding for each National Health Priority Area (NHPA) over time and by grant type, and to quantify the relationship between grants awarded and a range of measures of societal burden of disease (BoD). DESIGN AND SETTING: We conducted a retrospective analysis of NHMRC funding for each NHPA from 2000 to 2008 to assess the strength of correlation between level of NHMRC funding and contribution of each health condition to BoD. Information on mortality, incidence, prevalence, "healthy" years of life lost due to disability (YLD), years of life lost due to premature mortality (YLL) and disability-adjusted life-years (DALYs) was obtained from the 2003 Australian BoD study. Information on health system expenditure for each NHPA was obtained from an Australian Institute of Health and Welfare report. MAIN OUTCOME MEASURES: Observed versus expected number of grants; amount of funding allocated to each NHPA; relative contribution of each NHPA health condition to BoD. RESULTS: 6099 new and continuing NHMRC grants were linked to NHPAs. Total NHMRC funding by NHPA was strongly correlated with YLL and DALYs, but there was no clear association between the amount of funding per NHPA and YLD or health system expenditure. Based on the proportional contribution of each NHPA health condition to total NHPA-related DALYs, a higher than expected number of grants was allocated to diabetes and cancer research, and a lower than expected number to injury and mental health research. CONCLUSIONS: Some of Australia's NHPAs are better funded than others. The NHMRC could begin to redress this imbalance by allocating research and workforce development funding to less well developed research areas to ensure appropriate resourcing that is commensurate with their contribution to BoD. PMID- 20028296 TI - Simultaneous epidemics of influenza and malaria in the Australian Army in Palestine in 1918. AB - In October 1918, an Allied army (Egyptian Expeditionary Force) in Palestine experienced simultaneous epidemics of falciparum malaria and influenza during the cavalry campaign that defeated the Turkish Army. Malaria infection occurred 2 weeks after the advance of cavalry units into areas without environmental mosquito control. Pandemic influenza, now thought to be an A/H1N1 strain, struck at the same time. In the Egyptian Expeditionary Force of 315,000 soldiers, 773 died from malaria and 934 from influenza-pneumonia. Disease casualties outnumbered those due to combat by more than 37 to 1. Simultaneous infectious disease epidemics can cause mass casualties, capable of overwhelming any health service. PMID- 20028297 TI - Needlestick injury with smallpox vaccine. PMID- 20028298 TI - Charles Darwin's impressions of New Zealand and Australia, and insights into his illness and his developing ideas on evolution. AB - Charles Darwin visited New Zealand in December 1835, and Australia from January until March 1836, on the return portion of his voyage around the world in HMS Beagle. Despite the shortness of these visits, he retained an interest in these countries throughout his life, maintaining correspondence and receiving many biological specimens. His experiences in these places influenced his thinking on evolution, particularly on the evolution of man. Aspects of his health recorded during this part of the voyage support a new hypothesis for the diagnosis of the illness that Darwin endured for most of his life. PMID- 20028300 TI - Out of my depth in East Timor. PMID- 20028301 TI - Beware of laying blame! PMID- 20028303 TI - Christmas celebrates birth: new life, new love, new hope. PMID- 20028304 TI - Plague in a time of war: an experience in South Vietnam. PMID- 20028305 TI - Cough disorder: an allegory on DSM-IV. PMID- 20028306 TI - Relative radio-opacity of commonly consumed fish species in South East Queensland on lateral neck x-ray: an ovine model. AB - OBJECTIVE: To determine the relative radio-opacity on plain x-ray of bones of fish species commonly consumed in South East Queensland. DESIGN: A cadaveric sheep model was used to mimic the soft tissues of a human neck. Bones of 10 fish species were placed in the paratracheal tissues and adjacent to the larynx. X rays were taken and the images (including four control images with no bones) were incorporated into a Microsoft PowerPoint presentation to be interpreted by emergency specialists and registrars. Observers were blinded to which specimens contained fishbones and which did not. MAIN OUTCOME MEASURES: Sensitivity and specificity of plain x-rays for detecting impacted fishbones. RESULTS: Significant interobserver variability was identified. Despite this, the overall specificity of plain x-rays was 90%. The sensitivity of the technique was 79% overall, but varied significantly between fish species. CONCLUSION: Lateral soft tissue neck x-ray is an appropriate screening tool in cases of a suspected impacted fishbone. If a fishbone is identified on x-ray, the patient should be referred for endoscopy without further imaging. X-ray may be of limited value in cases of Dory or Spanish mackerel bone ingestion. In such cases, a computed tomography scan should be the first-line investigation. PMID- 20028307 TI - Fast versus slow bandaid removal: a randomised trial. AB - OBJECTIVE: To determine whether slow or fast bandaid removal is less painful. DESIGN, SETTING AND PARTICIPANTS: A prospective, randomised, crossover trial was carried out at James Cook University, Townsville. Participants were healthy volunteers from Years 2 and 3 of the James Cook University medical school program. INTERVENTIONS: Medium-sized bandaids were applied bilaterally in three standard body locations and removed using slow and fast techniques. MAIN OUTCOME MEASURES: Pain scores were assessed using an 11-point verbal numeric pain scale. RESULTS: 65 participants were included in the study. The overall mean pain score for fast bandaid removal was 0.92 and for slow bandaid removal was 1.58. This represents a highly significant difference of 0.66 (P < 0.001). CONCLUSION: In young healthy volunteers, fast bandaid removal caused less pain than slow bandaid removal. PMID- 20028308 TI - "The moment is all we have": patients and visitors reflect on a staff Christmas choir. AB - OBJECTIVE: To examine how performances by the Staff Christmas Choir of the Peter MacCallum Cancer Centre ("Peter Mac") affected inpatients, outpatients and visitors in 2008. DESIGN, SETTING AND PARTICIPANTS: During the Christmas season 2008, the Peter Mac Staff Christmas Choir gave seven performances at the Centre. Locations included inpatient wards, outpatient waiting areas and a cafeteria. To assess their response to the choir, oncology inpatients, outpatients and visitors (including early-departing bystanders) were given anonymous, semi-structured questionnaires during and after performances. To analyse the responses, we used a constructivist research approach informed by grounded theory. MAIN OUTCOME MEASURES: Participants' descriptions of the choir's effects on them. RESULTS: Questionnaires were returned by 111 people. The performances were received favourably by 93.7% of respondents, including nine from Jewish, Hindu or atheist backgrounds. Many said the music aroused positive emotions and memories. Several described transformative thoughts and physical reactions, felt affirmed by the Christmas spirit or message, and/or appreciated the peaceful or enlivened and social atmosphere. The choir also elicited personal perspectives about Christmas and Judaism, and the importance of "enjoying the moment". Only three respondents (2.7%) reported adverse effects, relating to emotional and audible intrusiveness. CONCLUSIONS: The Staff Christmas Choir created a supportive and uplifting atmosphere for many oncology patients and their visitors. However, responses from people from non-Christian backgrounds were limited, and further investigation is warranted to extend our understanding of the effect of Christmas music in Australian public health settings. PMID- 20028309 TI - Temporary henna tattoos with long-term consequences. PMID- 20028310 TI - The pressure of time. PMID- 20028311 TI - When two tribes go to war. PMID- 20028312 TI - A finger in the duodenum. PMID- 20028313 TI - The dark side of the moon. AB - OBJECTIVE: The belief that the full moon and disturbed behaviour are closely linked is alive and well, despite studies to the contrary. We investigated the possibility that there is an association between only extreme behavioural disturbance and the full moon. DESIGN, SETTING AND PARTICIPANTS: We undertook an observational study of patients with violent and acute behavioural disturbance who presented to the emergency department of Calvary Mater Newcastle and patients with less severe behaviour for whom hospital security calls were made. MAIN OUTCOME MEASURE: Proportion of patients for whom presentation or security call occurred in each lunar phase, modelled as a Poisson process. RESULTS: Of 91 patients with violent and acute behavioural disturbance, 21 (23%) presented during the full moon--double the number for other lunar phases (P = 0.002). Sixty (66%) had either alcohol intoxication or psychostimulant toxicity, and five attacked staff (biting [2], spitting [1], kicking [1] and scratching [1]). In contrast, 512 hospital security calls for patients with less severe behaviour were evenly distributed throughout the lunar cycle. CONCLUSION: Violent and acute behavioural disturbance manifested more commonly during the full moon. PMID- 20028314 TI - Successful resuscitation after drowning in a home swimming pool. PMID- 20028315 TI - Switched on. PMID- 20028317 TI - In silico approaches in G protein-coupled receptors (GPCRs) drug discovery: quo vadis? PMID- 20028316 TI - Rhodopsin and the others: a historical perspective on structural studies of G protein-coupled receptors. AB - The role of rhodopsin as a structural prototype for the study of the whole superfamily of G protein-coupled receptors (GPCRs) is reviewed in an historical perspective. Discovered at the end of the nineteenth century, fully sequenced since the early 1980s, and with direct three-dimensional information available since the 1990s, rhodopsin has served as a platform to gather indirect information on the structure of the other superfamily members. Recent breakthroughs have elicited the solution of the structures of additional receptors, namely the beta(1)- and beta(2)-adrenergic receptors and the A(2A) adenosine receptor, now providing an opportunity to gauge the accuracy of homology modeling and molecular docking techniques and to perfect the computational protocol. Notably, in coordination with the solution of the structure of the A(2A) adenosine receptor, the first "critical assessment of GPCR structural modeling and docking" has been organized, the results of which highlighted that the construction of accurate models, although challenging, is certainly achievable. The docking of the ligands and the scoring of the poses clearly emerged as the most difficult components. A further goal in the field is certainly to derive the structure of receptors in their signaling state, possibly in complex with agonists. These advances, coupled with the introduction of more sophisticated modeling algorithms and the increase in computer power, raise the expectation for a substantial boost of the robustness and accuracy of computer aided drug discovery techniques in the coming years. PMID- 20028320 TI - Customizing G Protein-coupled receptor models for structure-based virtual screening. AB - This review will focus on the construction, refinement, and validation of G Protein-coupled receptor models for the purpose of structure-based virtual screening. Practical tips and tricks derived from concrete modeling and virtual screening exercises to overcome the problems and pitfalls associated with the different steps of the receptor modeling workflow will be presented. These examples will not only include rhodopsin-like (class A), but also secretine-like (class B), and glutamate-like (class C) receptors. In addition, the review will present a careful comparative analysis of current crystal structures and their implication on homology modeling. The following themes will be discussed: i) the use of experimental anchors in guiding the modeling procedure; ii) amino acid sequence alignments; iii) ligand binding mode accommodation and binding cavity expansion; iv) proline-induced kinks in transmembrane helices; v) binding mode prediction and virtual screening by receptor-ligand interaction fingerprint scoring; vi) extracellular loop modeling; vii) virtual filtering schemes. Finally, an overview of several successful structure-based screening shows that receptor models, despite structural inaccuracies, can be efficiently used to find novel ligands. PMID- 20028319 TI - Progress in elucidating the structural and dynamic character of G Protein-Coupled Receptor oligomers for use in drug discovery. AB - G Protein-Coupled Receptors (GPCRs) are the most targeted group of proteins for the development of therapeutic drugs. Until the last decade, structural information about this family of membrane proteins was relatively scarce, and their mechanisms of ligand binding and signal transduction were modeled on the assumption that GPCRs existed and functioned as monomeric entities. New crystal structures of native and engineered GPCRs, together with important biochemical and biophysical data that reveal structural details of the activation mechanism(s) of this receptor family, provide a valuable framework to improve dynamic molecular models of GPCRs with the ultimate goal of elucidating their allostery and functional selectivity. Since the dynamic movements of single GPCR protomers are likely to be affected by the presence of neighboring interacting subunits, oligomeric arrangements should be taken into account to improve the predictive ability of computer-assisted structural models of GPCRs for effective use in drug design. PMID- 20028318 TI - Unraveling the structure and function of G protein-coupled receptors through NMR spectroscopy. AB - G protein-coupled receptors (GPCRs) are a large superfamily of signaling proteins expressed on the plasma membrane. They are involved in a wide range of physiological processes and, therefore, are exploited as drug targets in a multitude of therapeutic areas. In this extent, knowledge of structural and functional properties of GPCRs may greatly facilitate rational design of modulator compounds. Solution and solid-state nuclear magnetic resonance (NMR) spectroscopy represents a powerful method to gather atomistic insights into protein structure and dynamics. In spite of the difficulties inherent the solution of the structure of membrane proteins through NMR, these methods have been successfully applied, sometimes in combination with molecular modeling, to the determination of the structure of GPCR fragments, the mapping of receptor ligand interactions, and the study of the conformational changes associated with the activation of the receptors. In this review, we provide a summary of the NMR contributions to the study of the structure and function of GPCRs, also in light of the published crystal structures. PMID- 20028321 TI - G Protein-Coupled Receptors: target-based in silico screening. AB - In silico (or virtual) screening has become a common practice in current computer aided drug design efforts. However, application to hit discovery in the G Protein Coupled Receptors (GPCRs) arena was until recently hampered by the paucity of crystal structures available for this important class of pharmaceutical targets, forcing practitioners in the field to rely on GPCR models derived either ab initio or through homology modeling approaches. In this work we describe the EPIX in silico screening workflow which consists of the following stages: (1) Target modeling; (2) Preparation of screening library; (3) Docking; (4) Binding mode selection; (5) Scoring; (6) Consensus scoring and (7) Selection of virtual hits. This workflow was applied to the virtual screening of 13 GPCRs (5 biogenic amine receptors, 5 peptide receptors, 1 lipid receptor, 1 purinergic receptor and 1 cannabinoid receptor). Hit rates vary between 4% and 21% with higher hit rates usually obtained for biogenic amines and lower hits rates for peptide receptors. These data are analyzed in the context of the available experimental information (i.e., mutational data), the model (i.e., binding site location, and type of interactions) and the screening library. Specific examples are discussed in more detail as well as the future directions and challenges of this approach to in silico screening. PMID- 20028322 TI - Human A3 adenosine receptor as versatile G protein-coupled receptor example to validate the receptor homology modeling technology. AB - The development of ligands for the A(3) adenosine receptor (AR) has been directed mainly by traditional medicinal chemistry, but the influence of structure-based approaches is increasing. Rhodopsin-based homology modeling had been used for many years to obtain three-dimensional models of the A(3)AR, and different A(3)AR models have been published describing the hypothetical interactions with known A(3)AR ligands having different chemical scaffolds. The recently published structure of the human A(2A)AR provides a new template for GPCR modeling, however even use of the A(2A)AR as a template for modeling other AR subtypes is still imprecise. The models compared here are based on bovine rhodopsin, the human beta(2)-adrenergic receptor, and the A(2A)AR as templates. The sequence of the human A(3)AR contains only one cysteine residue (Cys166) in the second extracellular loop (EL2), which putatively forms a conserved disulfide bridge with the respective cysteine residues of TM3 (Cys83). Homology models of the A(3)AR have been helpful in providing structural hypotheses for the design of new ligands. Site-directed mutagenesis of the A(3)AR shows an important role in ligand recognition for specific residues in TM3, TM6 and TM7. PMID- 20028323 TI - Percutaneous radiofrequency ablation for unresectable colorectal liver metastases: time for shadows to disperse. AB - BACKGROUND: Despite the advent of combination chemotherapy regimens achieved within the last decade, long term survival of patients with unresectable metastatic disease from colorectal cancer remains poor. Thermal ablation procedures, including radiofrequency ablation (RFA), are considered feasible options in treating unresectable hepatic tumors either primary (hepatocellular carcinoma) or metastatic, the latter mainly arising from colorectal cancer. Percutaneous access is the least invasive RFA procedure. METHODS & RESULTS: A MEDLINE review unfolded a significant clinical heterogeneity among published series reporting on percutaneous RFA in hepatic metastatic disease from colorectal cancer, regarding study population, optimal time and treatment schemes pre- and post-RFA intervention. Notwithstanding, percutaneous RFA survival figures were consistently better than front line chemotherapy. Furthermore, a pooled analysis of larger series demonstrated a clear benefit in overall survival (HR 0.51, 95% CI 0.44 to 0.58). CONCLUSION: Albeit optimal indications are still pending, percutaneous RFA should nonetheless be considered a viable option in patients with unresectable metastatic disease, as it may prolong survival rates achieved with standard chemotherapy. PMID- 20028324 TI - Accelerated hypofractionated radiotherapy schedules in breast cancer: a review of the current literature. AB - Lumpectomy followed by breast irradiation is an alternative to mastectomy for patients with early-stage breast cancer. The purpose of radiation treatment following lumpectomy is to minimize the risk of recurrent cancer in the treated breast with as little toxicity as possible so that good cosmesis and function are maintained. Conventional fractionation schedules for postlumpectomy radiotherapy give 50 Gy in 2 Gy daily fractions, often with an additional boost to the tumor bed, resulting in treatment being given over 5-7 weeks. Delivering postoperative radiotherapy in a shorter period of time, provided it is as effective as longer treatment regiments, could result in greater convenience for patients. Moreover, given the high incidence of breast cancer, the use of a shorter fractionation schedule would decrease waiting lists in busy radiotherapy departments. We searched the medline (pubmed) and we reviewed all the relevant publications. We concluded that the accelerated hypofractionated schedules are safe in terms of cosmesis and effective in terms of local control. PMID- 20028325 TI - Is there a benefit from the concomitant use of immunosupression with anti-TNF in Crohn's disease; heads or tails? AB - Over the last some years the increasing knowledge on the pathogenesis of Crohn's disease led to the development of a number of biological agents targeting specific molecules involved in gut inflammation, first of all TNF-alpha and its receptors. Infliximab, adalimumab and certolizumab have been successful in inducing and maintaining remission in Crohn's disease at both short and long term. This was recently confirmed by a Cochrane meta-analysis and also open label extension follow-up and cohort studies. Emerging new data however indicate that combination therapy with infliximab-azathioprine appears to have added benefit in inducing steroid-free remission and mucosal healing than either infliximab or azathioprine alone in azathioprine-naive patients with early disease. Similarly the combination of steroids induction and infliximab was efficacious in luminal Crohn's disease. In contrast, there seems to be no synergism between methotrexate and infliximab. It is also less clear whether it is beneficial to use short or long-term infliximab-azathioprine combination in patients who previously failed therapy with azathioprine. In contrast, combination may potentially be associated with increased risk for infection and cancer. In case control-studies, especially the combination of steroids and anti-TNF and older age increased the risk for infectious complications, while scattered case reports point to the potentially increased risk of a rare form of non-Hodgkin's lymphoma (Hepatosplenic T cell lymphoma) with the use of azathioprine-anti-TNF combination. The aim of this review is to summarize the benefits and risks for the use combination therapy with TNF-alpha inhibitors in the treatment of Crohn's disease. PMID- 20028326 TI - Clinical trials of the past decade in the management of chronic kidney disease. AB - The three main causes of Chronic Kidney Diseases [CKD] are diabetes mellitus, chronic glomerulonephritis and hypertension. CKD is an increasing burden in the community as more patients fall prey to kidney failure. Both dialysis and renal transplantation are expensive modalities of treatment for end stage renal failure [ESRF]. Through the years many clinical trials have been performed to retard the progression of CKD to ESRF. Most of the trials focus on three main strategies which aim at renal retardation, namely, control of hypertension, treatment of proteinuria and control of hypercholesterolaemia. More recently, investigators have been exploring the role of high dose ARBs as well as the use of Aliskiren, a renin inhibitor. Early therapeutic intervention is necessary as it will contribute to better chances of minimising glomerular damage and in the case of some, even lead to the improvement of renal function with regression of glomerulosclerosis. PMID- 20028327 TI - The use of bevacizumab and trastuzumab beyond tumor progression: a new avenue in cancer treatment? AB - The use of the Monoclonal Antibodies (MoAbs) Bevacizumab (B) and Trastuzumab (T) beyond clinical progression in colorectal and breast cancer treatment is among the hottest topics in today's clinical oncology. Both observational and prospective studies, based on a sound preclinical basis, seem to support the notion that, simply replacing the cytotoxic drugs combined with the two MoAbs would provide an additional clinical benefit without stopping the biological agent. The aim of this review is to provide a critical analysis of the available clinical data, while waiting for the confirmatory prospective clinical trials still ongoing. The strength and the weakness of this innovative strategy, as well as the associated expense and toxicity issues will be discussed. PMID- 20028328 TI - Chia (Salvia hispanica): a systematic review by the natural standard research collaboration. AB - OBJECTIVE: To evaluate the scientific evidence on chia (Salvia hispanica) including history, folkloric precedent, expert opinion, pharmacology, dosing, interactions, adverse effects, and toxicology. This review serves as a clinical support tool. METHODS: Electronic searches were conducted in ten databases, 20 additional journals (not indexed in common databases), and bibliographies from 50 selected secondary references. No restrictions were placed on language or quality of publications. All literature collected pertained to efficacy in humans, dosing, precautions, adverse effects, use in pregnancy/lactation, interactions, alteration of laboratory assays, and mechanisms of action. Standardized inclusion/exclusion criteria are utilized for selection. Grades were assigned using an evidence-based grading rationale. RESULTS: The available human and non human studies show possible effectiveness for allergies, angina, athletic performance enhancement, cancer, coronary heart disease (CHD), heart attack, hormonal/endocrine disorders, hyperlipidemia, hypertension, stroke, and vasodilatation. Some evidence also suggests possible anticoagulant, antioxidant, and antiviral effects of Salvia hispanica. CONCLUSION: There is limited evidence supporting the efficacy of Salvia hispanica for any indication; thus far, only two clinical studies have examined the effects of Salvia hispanica on cardiovascular disease (CVD) risk factors (including body weight). One study showed some effects on some CVD risk factors, while the other did not. Neither study showed any effects of Salvia hispanica on weight loss. However, the historical use of Salvia hispanica suggests that it is safe for consumption by nonallergic individuals. Further rigorous examination is warranted pertaining to the use of Salvia hispanica as a dietary supplement, as well as in the treatment or prevention of human disease. PMID- 20028329 TI - Rapid sequence intubation: a review of recent evidences. AB - Rapid sequence intubation is an essential bullet in the maintenance of patency of the airway during intubation in emergency. It is a valid method in all those situations where you can not determine whether the patient is fasting or not. But RSI is not applicable in all critically ill patients. The presence of severe acidosis, depletion of intravascular volume, heart failure and severe pulmonary disease may complicate the pre-induction period as the induction, leading to the onset of vasodilatation and hypotension. Another complication is represented by Hypoxemia during the manoeuvre. The algorithm of RSI consists in six steps: pre oxygenation, premedication, myo-relaxation and induction, intubation, primary and secondary confirmation, post-intubation patient management. Propofol has replaced Thiopental as the most common intravenous ipnotic. In hypotensive patients Ketamine represents a viable alternative. Succinylcholine is the most common neuromuscular relaxant used in the RSI. The not depolarizing NMBAs are an alternative to Succinylcholine. Among these, the most important is the Rocuronium. This drug is characterized by a fairly rapid onset (1-2 min) and an intermediate half-life (45-70 min). The onset depends on the dosage used. The problem that limits the use of Rocuronium is the fact that its duration of action is much longer than that of Succinylcholine, especially when used at higher doses. This problem can be solved through the use of Sugammadex. As a muscle relaxant chelating Sugammadex antagonizes the effects induced by Rocuronium on muscle tissue and quickly resolve the blockade. PMID- 20028330 TI - Emerging antiviral drugs for hepatitis C virus. AB - Infection with hepatitis C virus (HCV) is a global health problem that affects approximately 170 million people worldwide. The current standard therapy with peginterferon alpha plus ribavirin for 48 weeks results in a sustained virologic response in less than 50% of patients with chronic hepatitis C genotype 1--the most prevalent type of HCV in North America and Europe. Development of new antiviral medicines has been hampered by the lack of an effective cell culture system and small-animal model. Herein we review recent progress in the development of new treatments under investigation in clinical trials, including specifically targeted antiviral therapy for HCV such as NS3/4A protease and NS5B polymerase inhibitors. PMID- 20028331 TI - Spectrum of benign lesions mimicking a malignant stricture at the liver hilum. AB - There is a broad spectrum of benign disorders of the biliary system that resemble hilar cholangiocarcinoma (HCCA) in terms of clinical, pathologic, and imaging findings. No unifying features were found to characterize patients with benign hilar obstruction and distinguish these patients from those with cholangiocarcinoma. Imaging plays a vital role in aiding the differentiation of benign and malignant disease, defining the location and extent of the process, as well as directing biopsy. However, even when lesions at the liver hilum are detected with the highest sensitivity, none of the imaging modalities can reliably characterize and confirm the underlying type of disease. Excessive reliance on cholangiographic or endoscopic biopsy results is dangerous, because tissue sampling is not always diagnostic and a potentially resectable malignancy can be overlooked. Therefore, the preferred treatment option to patients with suspicious hilar lesions should remain resection for presumed malignancy. Local resection with adequate reconstruction excludes a malignant lesion, and provides means of biliary decompression with low mortality and morbidity rate. PMID- 20028332 TI - Parallel conduction of the phase I preventive and therapeutic trials based on the Tat vaccine candidate. AB - The native HIV-1 Tat protein was chosen as vaccine candidate for phase I clinical trials in both uninfected (ClinicalTrials.gov identifier: NCT00529698) and infected volunteers (ClinicalTrials.gov identifier: NCT00505401). The rationale was based on the role of Tat in the natural infection and AIDS pathogenesis, on the association of Tat-specific immune responses with the asymptomatic stage and slow-progression rate as well as on its sequence conservation among HIV clades (http://www.hiv1tat-vaccines.info/). The parallel conduction in the same clinical centers of randomized, double blind, placebo-controlled phase I studies both in healthy, immunologically competent adults and in HIV-infected, clinically asymptomatic, individuals represents a unique occasion to compare the vaccine induced immune response in both the preventive and therapeutic setting. In both studies, the same lot of the native Tat protein was administered 5 times, every four weeks, subcute (SC) with alum adjuvant or intradermic (ID), in the absence of adjuvant, at 7.5 microg, 15 microg or 30 microg doses, respectively. The primary and secondary endpoints of these studies were the safety and immunogenicity of the vaccine candidate, respectively. The study lasted 52 weeks and monitoring was conducted for on additional 3 years. The results of both studies indicated that the Tat vaccine is safe and well tolerated both locally and systemically and it is highly immunogenic at all the dosages and by both routes of administration. Vaccination with Tat induced a balanced immune response in uninfected and infected individuals. In particular, therapeutic immunization induced functional antibodies and partially reverted the marked Th1 polarization of anti-Tat immunity seen in natural infection, and elicited a more balanced Th1/Th2 immune response. Further, the number of CD4 T cells correlated positively with anti-Tat antibody titers. Based on these results, a phase II study is ongoing in infected drug-treated individuals (http://www.hiv1tat-vaccines.info/). PMID- 20028333 TI - Treatment of bone metastases in lung cancer: the actual role of zoledronic acid. AB - Approximately 30-40% of NSCLC patients develop bone metastases. Bone metastases are associated with a significant increase in skeletal-related events (SREs), including severe bone pain, hypercalcemia, pathological fractures, spinal cord compression. These SREs result in impaired mobility, reduced quality of life, and frequently require therapeutic intervention (radiation therapy, surgery and systemic treatments). The normal balance of formation of new bone by osteoblasts and the resorption of old bone by osteoclasts becomes imbalanced and/or uncoupled, leading to the development of lesions that are osteolytic, osteoblastic, or a combination of both. The current National Comprehensive Cancer Network (NCCN) Clinical Practice Giudelines in Oncology recommend palliative external-beam radiotherapy for the treatment of bone metastases in patients with NSCLC and healthcare professionals treating such patients are urged to consider bisphosphonate therapy. Zoledronic acid is the first and only bisphosphonate that has proven efficacy for the treatment of bone metastases from a broad range of solid tumor types, including lung cancer. PMID- 20028334 TI - Immune cell entry to central nervous system--current understanding and prospective therapeutic targets. AB - Under normal physiological conditions there is minimal entry of immune cells into the central nervous system (CNS) for the purpose of immune surveillance. During inflammation, however, extensive infiltration of immune cells can lead to the induction of CNS autoimmune disease, for example multiple sclerosis (MS) and its animal model experimental autoimmune encephalomyelitis (EAE). The barriers that regulate cellular entry are the blood-brain barrier (BBB) within the CNS parenchyma, and the blood-cerebrospinal fluid (blood-CSF) barrier within the choroid plexus. Understanding how these barriers function to allow the passage of leukocytes from the periphery into the CNS for normal immune surveillance, and under inflammatory conditions, is vital for the development of novel therapeutics targeting immune cell migration in CNS diseases. Contributions from selectins, chemokines, integrins and matrix metalloproteinases allow the migration of leukocytes across the BBB and into the CNS parenchyma. In EAE and MS, the strict maintenance of this process is lost and a large influx of cells is seen. This review focuses on the role of these homing molecules, chemokines and enzymes in the entry of leukocytes into the CNS during inflammatory conditions. It concludes with a model of immune cell entry of the CNS, summarising the current knowledge in this area. Targeting specific molecules to prevent infiltration of inflammatory cells into the CNS could allow disease inhibition without compromising beneficial immune surveillance. PMID- 20028335 TI - Ref2, a regulatory subunit of the yeast protein phosphatase 1, is a novel component of cation homoeostasis. AB - Maintenance of cation homoeostasis is a key process for any living organism. Specific mutations in Glc7, the essential catalytic subunit of yeast protein phosphatase 1, result in salt and alkaline pH sensitivity, suggesting a role for this protein in cation homoeostasis. We screened a collection of Glc7 regulatory subunit mutants for altered tolerance to diverse cations (sodium, lithium and calcium) and alkaline pH. Among 18 candidates, only deletion of REF2 (RNA end formation 2) yielded increased sensitivity to these conditions, as well as to diverse organic toxic cations. The Ref2F374A mutation, which renders it unable to bind Glc7, did not rescue the salt-related phenotypes of the ref2 strain, suggesting that Ref2 function in cation homoeostasis is mediated by Glc7. The ref2 deletion mutant displays a marked decrease in lithium efflux, which can be explained by the inability of these cells to fully induce the Na+-ATPase ENA1 gene. The effect of lack of Ref2 is additive to that of blockage of the calcineurin pathway and might disrupt multiple mechanisms controlling ENA1 expression. ref2 cells display a striking defect in vacuolar morphogenesis, which probably accounts for the increased calcium levels observed under standard growth conditions and the strong calcium sensitivity of this mutant. Remarkably, the evidence collected indicates that the role of Ref2 in cation homoeostasis may be unrelated to its previously identified function in the formation of mRNA via the APT (for associated with Pta1) complex. PMID- 20028336 TI - Selective inhibition of beta-F1-ATPase mRNA translation in human tumours. AB - Down-regulation of beta-F1-ATPase (the catalytic subunit of the mitochondrial H+ ATP synthase) is a hallmark of many human tumours. The expression level of beta F1-ATPase provides a marker of the prognosis of cancer patients, as well as of the tumour response to chemotherapy. However, the mechanisms that participate in down-regulating its expression in human tumours remain unknown. In the present study, we have investigated the expression of beta-F1-ATPase mRNA (termed beta mRNA) in breast, colon and lung adenocarcinomas and squamous carcinomas of the lung. Despite the down-regulation of the protein, tumour beta-mRNA levels remained either unchanged (breast and lung adenocarcinomas) or significantly increased (colon and squamous lung carcinomas) when compared with paired normal tissues, suggesting a specific translation-masking event for beta-mRNA in human cancer. Consistently, we show using cell-free translation assays that a large fraction (approximately 70%) of protein extracts derived from breast and lung adenocarcinomas specifically repress the translation of beta-mRNA. We show that the 3'UTR (3' untranslated region) of human beta-mRNA is a relevant cis-acting element required for efficient translation of the transcript. However, an RNA chimaera bearing the 3'UTR of human beta-mRNA does not recapitulate the inhibitory effect of tumour extracts on beta-mRNA translation. Overall, the findings of the present study support the hypothesis that down-regulation of the bioenergetic activity of mitochondria in human tumours is exerted by translation silencing of beta-mRNA. PMID- 20028337 TI - Band 3 Edmonton I, a novel mutant of the anion exchanger 1 causing spherocytosis and distal renal tubular acidosis. AB - dRTA (distal renal tubular acidosis) and HS (hereditary spherocytosis) are two diseases that can be caused by mutations in the gene encoding the AE1 (anion exchanger 1; Band 3). dRTA is characterized by defective urinary acidification, leading to metabolic acidosis, renal stones and failure to thrive. HS results in anaemia, which may require regular blood transfusions and splenectomy. Mutations in the gene encoding AE1 rarely cause both HS and dRTA. In the present paper, we describe a novel AE1 mutation, Band 3 Edmonton I, which causes dominant HS and recessive dRTA. The patient is a compound heterozygote with the new mutation C479W and the previously described mutation G701D. Red blood cells from the patient presented a reduced amount of AE1. Expression in a kidney cell line showed that kAE1 (kidney AE1) C479W is retained intracellularly. As kAE1 is a dimer, we performed co-expression studies and found that, in kidney cells, kAE1 C479W and G701D proteins traffic independently from each other despite their ability to form heterodimers. Therefore the patient carries one kAE1 mutant that is retained in the Golgi (G701D) and another kAE1 mutant (C479W) located in the endoplasmic reticulum of kidney cells, and is thus probably unable to reabsorb bicarbonate into the blood. We conclude that the C479W mutant is a novel trafficking mutant of AE1, which causes HS due to a decreased cell-surface AE1 protein and results in dRTA due to its intracellular retention in kidney. PMID- 20028338 TI - Creutzfeldt-Jakob disease with PRNP G114V mutation in a Chinese family. AB - BACKGROUND: Recent evidence has shown clinical phenotypic heterogeneity of inherited prion diseases, even between patients harbouring the same mutation in the PRNP gene. OBJECTIVE AND METHODS: We collected clinical data from a Chinese family with autosomal dominant dementia and screened the PRNP gene on 28 living members. A stereotactic biopsy of the right frontal lobe of the proband was performed. RESULTS: The family comprised four affected individuals within two successive generations. The age of onset was in 30 or 40 s, and the duration was about 2-3 years. Clinical features of the affected members included neuropsychiatric disturbances, progressive dementia and extrapyramidal symptoms. Immunostaining for prion protein showed fine granular deposits of PrP(sc) in the neuropil. The PRNP gene analysis demonstrated a heterozygous G114V mutation in 15 family members. The proband was diagnosed as familial Creutzfeldt-Jakob disease (fCJD). CONCLUSION: This study strengthens the linkage of the G114V mutation to CJD. It supports the worldwide distribution of fCJD despite differences in genetic background. PMID- 20028339 TI - Acute encephalitis with refractory, repetitive partial seizures (AERRPS): a peculiar form of childhood encephalitis. AB - OBJECTIVE: We conducted a nationwide multicenter study in Japan to elucidate the clinical and laboratory characteristics of acute encephalitis with refractory, repetitive partial seizures (AERRPS). MATERIALS AND METHODS: Clinical and laboratory features, treatment, and outcome were assessed using a structured questionnaire. RESULTS: Twenty-nine children were enrolled in the study. Refractory and repetitive partial seizures accompanied by fever were the cardinal clinical features. Partial seizures consisted principally of eye deviation or facial twitching, being periodically repeated during the acute phase. These seizures were refractory to conventional anticonvulsants and were only suppressed by high-dose intravenous barbiturate administration. Rhythmic activities on electroencephalography and non-specific cerebral atrophy on neuroimaging were common. Serum or cerebrospinal antibodies against GluRepsilon2 were positive in six patients. General prognosis was unfavorable due to intractable epilepsy and cognitive deficits. CONCLUSION: Based on the peculiar and homogenous features, AERRPS can be regarded as a distinct clinical entity. PMID- 20028340 TI - Well-being in patients with amyotrophic lateral sclerosis and their next of kin over time. AB - OBJECTIVES: The well-being and physical function among patients with ALS and their next of kin was studied over time. MATERIALS AND METHODS: Thirty-five patients with ALS and their next of kin were studied with respect to physical, general and psychological well-being by the visual analogue scale (VAS) every 4-6 months. Physical function in patients was rated by the ALSFRS-R and the Norris scale. Patients and next of kin rated the well-being of themselves and their counterpart. RESULTS: The well-being was stable and there was a relation between the well-being of patients and next of kin throughout the time studied. Next of kin rated the well-being of the patients worse than patients rated themselves, while patients rated the well-being of their next of kin at the same level as their counterpart. CONCLUSIONS: The basic state of well-being as well as the interaction between patient and next of kin seem to be factors that influence the well-being of both patients and their next of kin. PMID- 20028341 TI - Clinical, neuroimaging and neurophysiological features in addicts with manganese ephedrone exposure. AB - OBJECTIVE: To identify biomarkers supporting the clinical diagnosis of manganism in patients several years after exposure to manganese (Mn). METHODS: Neurophysiological examinations, magnetic resonance imaging (MRI), single-photon emission computed tomography and fluorodeoxyglycose (FDG) positron emission tomography were performed in four former ephedrone addicts with extrapyramidal symptoms. RESULTS: Peripheral nervous system was not affected. No patients had reduced uptake of (123)I Ioflupane in the striatum. MRI signal intensities were slightly changed in the basal ganglia. All patients showed a widespread, but not uniform, pathological pattern of FDG uptake with changes mainly located to the central part of the brain including the basal ganglia and the surrounding white matter. CONCLUSIONS: Presynaptic neurons in the nigrostriatal pathway are intact in Mn-induced parkinsonism after prolonged abstinence from ephedrone. The diagnosis is principally based on clinical signs and the history of drug abuse. PMID- 20028342 TI - The effect of sildenafil citrate (Viagra) on cerebral blood flow in patients with cerebrovascular risk factors. AB - OBJECTIVES: Sildenafil citrate is widely used for erectile dysfunction. The present study examined the short-term effects of sildenafil administration in individuals with cerebrovascular risk factors, including patients with a history of stroke. MATERIALS AND METHODS: Twenty-five consecutive male patients with erectile dysfunction and vascular risk factors were included in the study. A perfusion brain SPECT study was performed at baseline and 1 h after the oral administration of sildenafil. RESULTS: Associations between any of the risk factors and the perfusion scores were not detected, with the exception of stroke. Stroke patients showed significantly more areas with diminished perfusion after sildenafil administration compared to baseline. CONCLUSIONS: In patients with diabetes or hypertension, a dose of 50 mg sildenafil does not appear to produce detrimental effects on cerebral blood flow. However, patients with a history of stroke may be at increased risk of hemodynamic impairment after the use of sildenafil. PMID- 20028343 TI - Leptin and ghrelin concentrations and weight loss in Parkinson's disease. AB - OBJECTIVES: To investigate the role of leptin, ghrelin, GH and IGF-1 in energy balance disturbances in Parkinson's disease (PD). MATERIALS AND METHODS: Thirty nine patients were included: 11 PD patients with unintentional weight loss, 16 PD patients without weight loss and 12 controls. UPDRS, MMSE, MADRS, appetite scale, BMI, adipose tissue content, plasma leptin and active ghrelin concentrations and serum GH, IGF-1, TSH, T3 and T4, concentrations were evaluated. RESULTS: A lower plasma leptin concentration and a higher serum IGF-1 concentration were found in PD patients with weight loss. BMI and the content of adipose tissue were positively correlated with leptin concentration in all PD patients. Paradoxically, the lower BMI was, the lower plasma active ghrelin concentration was in PD patients with the weight loss. CONCLUSION: These findings confirm that changes of plasma leptin concentration occur in PD patients with loss of weight. PMID- 20028344 TI - Apraxia related with subcortical lesions due to cerebrovascular disease. AB - OBJECTIVES: To examine whether ideomotor apraxia exists in patients with subcortical ischemic lesions. PATIENTS AND METHODS: A matched-control, prospective and multi-centered research design was used. Ideomotor apraxia, anxiety and depression were assessed by the Movement Imitation Test and the Hamilton scales, respectively. RESULTS: Forty two consecutive patients with subcortical ischemic stroke and an equal number of healthy participants, matched in age and sex were included. Paired-sample t-tests showed that patients had significantly more apractic elements in their movements (t = 5.03, P < 0.01), higher anxiety (t = -2.55, P = 0.0014) and depression levels (t = -2.61, P = 0.012) than their healthy matched participants. Participants with higher anxiety and depression scores performed worse on the Movement Imitation Test. CONCLUSIONS: Ischemic damage of subcortical modular systems may affect praxis. PMID- 20028345 TI - Ischaemic preconditioning is related to decreasing levels of extracellular adenosine that may be metabolically useful in the at-risk myocardium: an experimental study in the pig. AB - AIM: 'Pre-treatment' with short repetitive periods of ischaemia (ischaemic preconditioning) has proved to be a powerful mechanism for modification of the extent of myocardial damage following acute coronary artery occlusion. The exact mechanism of protection induced by ischaemic preconditioning is not known. We herewith put forward a contributing component for protection with preconditioning involving a shift in the adenylate kinase (AK) equilibrium reaction in favour of adenosine triphosphate (ATP) formation. METHODS: A coronary artery was occluded in anaesthetized thoracotomized pigs to induce ischaemic preconditioning as well as a longer period of ischaemia. Microdialysis probes were inserted in ischaemic and control myocardium and were infused with (14)C- adenosine with two different specific activities. (14)C-lactate was identified and measured in the effluent. RESULTS: (14)C-adenosine was taken up by non-preconditioned and preconditioned myocardium during ischaemia. Significantly increased levels of (14)C-lactate were recovered in preconditioned myocardium. (14)C-adenosine with high specific activity resulted in a specific activity of lactate that was 2.7 times higher than that of lactate after administration of (14)C-adenosine with low specific activity. Mass spectrography verified the identity of (14)C-lactate. CONCLUSIONS: Preconditioning up-regulates a new metabolic pathway (starting with 5' nucleotidase and ending up with lactate) resulting in ATP formation in the micromolar range on top of another effect terminating in a useful shift in the AK equilibrium reaction in favour of ATP generation in the millimolar range. Although the up-regulation of the purine nucleoside phosphorylase pathway is clearly demonstrated, its biological relevance remains to be proved. PMID- 20028346 TI - Angiotensin II regulates endothelial cell migration through calcium influx via T type calcium channel in human umbilical vein endothelial cells. AB - AIM: The T-type calcium channel is expressed in vascular endothelial cells, but its role in endothelial cell function is yet to be elucidated. We analysed the endothelial functional role of T-type calcium channel-dependent calcium under angiotensin II (Ang II) stimulation. METHODS: Human umbilical vein endothelial cells were co-incubated with hormone at 10(-7) m and either Efonidipine 10(-5) m or Verapamil 10(-5) m or Mibefradil 10(-5) m or Wortmannin 10(-6) m. The contribution of Ang II receptors was evaluated using PD123319 10(-7) m and ZD 7155 10(-7) m. The calcium ion concentration was observed using Fluo-3 acetossimetil ester. The cells were observed after 3, 6, 9 and 12 h. RESULTS: The microfluorescence method points out that Ang II induces intracellular calcium modulation in time by distinct mechanisms. AT2 receptor blockade is necessary to observe significant increase in [Ca(2+)](i) levels. Pre-treatment with Mibefradil abolishes Ang II -induced cell migration. CONCLUSIONS: Our data show that Ang II, via AT1 receptor, modulates calcium concentration involving T-type calcium channel and L-type calcium channel but only the calcium influx via T-type calcium channels regulates endothelial cell migration which is essential for angiogenesis. PMID- 20028348 TI - Moderate alcohol consumption aggravates high-fat diet induced steatohepatitis in rats. AB - BACKGROUND: Nonalcoholic steatohepatitis (NASH) develops in the absence of chronic and excessive alcohol consumption. However, it remains unknown whether moderate alcohol consumption aggravates liver inflammation in pre-existing NASH condition. METHODS: Sprague-Dawley rats were first fed ad libitum with Lieber DeCarli high-fat diet (71% energy from fat) for 6 weeks to induce NASH, as demonstrated previously. Afterwards, these rats were continuously fed with high fat diet (HFD, 55% total energy from fat) or high fat plus alcohol diet (HFA, 55% energy from fat and 16% energy from alcohol) for an additional 4 weeks. Pathological lesions including fat accumulation and inflammatory foci in liver were examined and graded. Lipid peroxidation and apoptotic hepatocytes in the liver were assessed. The mRNA expressions of tumor necrosis factor-alpha (TNFalpha) and TNF receptor 1 (TNF-R1), Fas death receptor (Fas) and Fas ligant (FasL), IL-1beta and IL-12 were determined by real-time PCR. Protein levels of total and cleaved caspase-3, CYP2E1, Bax, and Bcl-2 were measured by western blotting. RESULTS: The number of hepatic inflammatory foci and apoptotic hepatocytes were significantly increased in rats fed with HFA as compared with those in HFD-fed rats. The aggravated inflammatory response and cellular apoptosis mediated by HFA were associated with elevated mRNA expression of Fas/FasL and cleaved caspase-3 protein. Although no significant differences were observed between HFD and HFA groups, the levels of lipid peroxidation, Bax and Bcl-2 protein concentration, and mRNA levels of other inflammatory cytokines were significantly higher in these 2 groups than those in the control group. CONCLUSIONS: These data suggest that even moderate alcohol consumption can cause more hepatic inflammation and cellular apoptosis in a pre-existing NASH condition. PMID- 20028349 TI - Differential central NOS-NO signaling underlies clonidine exacerbation of ethanol evoked behavioral impairment. AB - BACKGROUND: The molecular mechanisms that underlie clonidine exacerbation of behavioral impairment caused by ethanol are not fully known. We tested the hypothesis that nitric oxide synthase (NOS)-derived nitric oxide (NO) signaling in the locus coeruleus (LC) is implicated in this phenomenon. METHODS: Male Sprague-Dawley rats with intracisternal (i.c.) and jugular vein cannulae implanted 6 days earlier were tested for drug-induced behavioral impairment. The latter was assessed as the duration of loss of righting reflex (LORR) and rotorod performance every 15 minutes until the rat recovered to the baseline walk criterion (180 seconds). In a separate cohort, we measured p-neuronal NOS (nNOS), p-endothelial NOS (eNOS), and p-ERK1/2 in the LC following drug treatment, vehicle, or NOS inhibitor. RESULTS: Rats that received clonidine [60 Ig/kg, i.v. (intravenous)] followed by ethanol (1 or 1.5 g/kg, i.v.) exhibited synergistic impairment of rotorod performance. Intracisternal pretreatment with nonselective NOS inhibitor N(omega)-nitro-L-arginine methyl ester (L-NAME, 0.5 mg) or selective nNOS inhibitor N-propyl-L-arginine (1 microg) exacerbated the impairment of rotorod performance caused by clonidine-ethanol combination. Exacerbation of behavioral impairment was caused by L-NAME enhancement of the effect of ethanol, not clonidine. L-NAME did not influence blood ethanol levels; thus, the interaction was pharmacodynamic. LORR caused by clonidine (60 microg/kg, i.v.)-ethanol (1 g/kg, i.v.) combination was abolished by selective inhibition of central eNOS (L-NIO, 10 microg i.c.) but not by nNOS inhibition under the same conditions. Western blot analyses complemented the pharmacological evidence by demonstrating that clonidine-ethanol combination inhibits phosphorylation (activation) of nNOS (p-nNOS) and increases the level of phosphorylated eNOS (p-eNOS) in the LC; the change in p-nNOS was paralleled by similar change in LC p-ERK1/2. NOS inhibitors alone did not affect the level of nitrate/nitrite, p-nNOS, p-eNOS, or p-ERK1/2 in the LC. CONCLUSIONS: Alterations in NOS-derived NO in the LC underlie clonidine-ethanol induced behavioral impairment. A decrease in nNOS activity, due at least partly to a reduction in nNOS phosphorylation, mediates rotorod impairment, while enhanced eNOS activity contributes to LORR, elicited by clonidine-ethanol combination. PMID- 20028350 TI - Revisiting intragastric ethanol intubation as a dependence induction method for studies of ethanol reward and motivation in rats. AB - BACKGROUND: The purpose of this study was to re-examine intragastric ethanol intubation as a dependence induction method that effectively induces physical dependence upon ethanol over a short time period, is devoid of intrinsic stress artifacts, inexpensive, and easy to implement. METHODS: Male Wistar rats were subjected to ethanol dependence induction via intragastric ethanol intubation. Ethanol solution (final concentration 20%, made up in a dietary liquid vehicle consisting of powdered milk, sucrose, and water) was intubated 4 times per day, at 4-hour intervals, for 6 consecutive days (for a total of 10 g/kg/day). The utility of this procedure was evaluated for inducing physical dependence, determined by daily and final withdrawal ratings. Anxiety-like behavior associated with ethanol dependence history was examined using the elevated plus maze (EPM) test, conducted 5 days after ethanol withdrawal. To evaluate whether potential stress-like effects of intragastric intubation per se produce lasting effects on behavior, experimentally naive rats were compared with vehicle intubated rats for anxiety-like behavior on the EPM. RESULTS: Blood alcohol levels reached stable levels between 200 and 250 mg%, measured 1 hour after the second and third ethanol intubation on days 2, 4, and 6. Ethanol-treated rats developed significant somatic withdrawal signs, recorded daily between 10 and 12 hours after the last ethanol administration. At 5 days postwithdrawal, ethanol treated rats showed significant anxiety-like behavior, measured by decreased open arm time and open arm entries on the EPM, compared with vehicle controls. Additionally, ethanol postdependent rats showed decreased open arm time compared with experimentally naive rats. EPM performance did not differ between vehicle intubated and naive rats. No withdrawal seizures were observed and mortality rate was near zero. CONCLUSIONS: These findings suggest that intragastric ethanol administration produces a behavioral profile consistent with ethanol dependence (i.e., significant withdrawal signs after termination of ethanol exposure and elevated anxiety-like behavior persisting beyond completion of physical withdrawal), and that the intubation procedure itself does not produce lasting nonspecific anxiety-like effects. Thus, under the conditions employed here, this procedure provides an effective tool for inducing and evaluating the consequences of ethanol dependence in animal models of ethanol reward and motivation. PMID- 20028351 TI - Feasibility of using interactive voice response to monitor daily drinking, moods, and relationship processes on a daily basis in alcoholic couples. AB - BACKGROUND: Daily process research on alcohol involvement has used paper-and pencil and electronic data collection methods, but no studies have yet tested the feasibility of using Interactive Voice Response (IVR) technology to monitor drinking, affective, and social interactional processes among alcoholic (ALC) couples. This study tested the feasibility of using IVR with n = 54 ALC couples. METHODS: Participants were n = 54 couples (probands who met criteria for a past 1 year alcohol use disorder and their partners) recruited from a substance abuse treatment center and the local community. Probands and their partners reported on their daily drinking, marital interactions, and moods once a day for 14 consecutive days using an IVR system. Probands and partners were on average 43.4 and 43.0 years old, respectively. RESULTS: Participants completed a total of 1,418 out of a possible 1,512 diary days for an overall compliance rate of 93.8%. ALC probands completed an average of 13.3 (1.0) diary reports, and partners completed an average of 13.2 (1.0) diary reports. On average, daily IVR calls lasted 7.8 (3.0) minutes for ALC probands and 7.6 (3.0) minutes for partners. Compliance was significantly lower on weekend days (Fridays and Saturdays) compared to other weekdays for probands and spouses. Although today's intoxication predicted tomorrow's noncompliance for probands but not spouses, the strongest predictor of proband's compliance was their spouse's compliance. Daily anxiety and marital conflict were associated with daily IVR nonresponse, which triggered automated reminder calls. CONCLUSIONS: Findings supported that IVR is a useful method for collecting daily drinking, mood, and relationship process data from alcoholic couples. Probands' compliance is strongly associated with their partners' compliance, and automated IVR calls may facilitate compliance on high anxiety, high conflict days. PMID- 20028352 TI - Phosphodiesterase type 4 inhibition does not restore ocular dominance plasticity in a ferret model of fetal alcohol spectrum disorders. AB - BACKGROUND: There is growing evidence that deficits in neuronal plasticity account for some of the neurological problems observed in fetal alcohol spectrum disorders (FASD). Recently, we showed that early alcohol exposure results in a permanent impairment in visual cortex ocular dominance (OD) plasticity in a ferret model of FASD. This disruption can be reversed, however, by treating animals with a Phosphodiesterase (PDE) type 1 inhibitor long after the period of alcohol exposure. AIM: Because the mammalian brain presents different types of PDE isoforms we tested here whether inhibition of PDE type 4 also ameliorates the effects of alcohol on OD plasticity. MATERIAL AND METHODS: Ferrets received 3.5 g/Kg alcohol i.p. (25% in saline) or saline as control every other day between postnatal day (P) 10 to P30, which is roughly equivalent to the third trimester equivalent of human gestation. Following a prolonged alcohol-free period (10 to 15 days), ferrets had the lid of the right eye sutured closed for 4 days and were examined for ocular dominance changes at the end of the period of deprivation. RESULTS: Using in vivo electrophysiology we show that inhibition of PDE4 by rolipram does not restore OD plasticity in alcohol-treated ferrets. CONCLUSION: This result suggests that contrary to PDE1, PDE4 inhibition does not play a role in the restoration of OD plasticity in the ferret model of FASD. PMID- 20028353 TI - Phosphatidylethanol and alcohol consumption in reproductive age women. AB - BACKGROUND: Fetal alcohol disorders are preventable, but self-reported alcohol consumption can be misleading and impede effective treatment. Biomarkers represent an alternative method for assessing alcohol use, and this study evaluated the relationship between blood phosphatidylethanol (PEth) and alcohol use in a sample of reproductive age women. METHODS: Alcohol use was estimated by validated self-report methods in 80 nonpregnant women ages 18 to 35. PEth was measured by a contracted laboratory using a liquid chromatography-tandem mass spectrometry assay. Regression methods appropriate for the distribution of PEth were used to define its relationship to alcohol consumption during the prior 2 weeks and explore the effects of drinking patterns on this association. Receiver operating characteristic analysis was used to estimate the sensitivity of PEth for various drinking levels at 95% specific cutoffs. RESULTS: PEth had a positive linear association with grams of alcohol consumed (p < 0.001), and was detectable in 93% of subjects consuming an average of 2 or more drinks per day. The relationship between total alcohol consumption and PEth may be stronger in women with recent heavy drinking days. The relationship between drinking and PEth varied considerably between individuals, and sensitivity for a certain amount of drinking was low at a highly specific cutoff concentration. CONCLUSIONS: PEth is a highly sensitive indicator of moderate and heavy alcohol consumption in reproductive age women and may complement the use of self-report alcohol screens when additional objective markers of alcohol use are desirable. However, choosing a highly valid cutoff concentration for PEth to differentiate various levels of alcohol consumption may not be feasible. PMID- 20028354 TI - Disruptions in functional network connectivity during alcohol intoxicated driving. AB - BACKGROUND: Driving while under the influence of alcohol is a major public health problem whose neural basis is not well understood. In a recently published functional magnetic resonance imaging (fMRI) study (Meda et al., 2009), our group identified 5, independent critical driving-associated brain circuits whose inter regional connectivity was disrupted by alcohol intoxication. However, the functional connectivity between these circuits has not yet been explored in order to determine how these networks communicate with each other during sober and alcohol-intoxicated states. METHODS: In the current study, we explored such differences in connections between the above brain circuits and driving behavior, under the influence of alcohol versus placebo. Forty social drinkers who drove regularly underwent fMRI scans during virtual reality driving simulations following 2 alcohol doses, placebo and an individualized dose producing blood alcohol concentrations (BACs) of 0.10%. RESULTS: At the active dose, we found specific disruptions of functional network connectivity between the frontal temporal-basal ganglia and the cerebellar circuits. The temporal connectivity between these 2 circuits was found to be less correlated (p < 0.05) when driving under the influence of alcohol. This disconnection was also associated with an abnormal driving behavior (unstable motor vehicle steering). CONCLUSIONS: Connections between frontal-temporal-basal ganglia and cerebellum have recently been explored; these may be responsible in part for maintaining normal motor behavior by integrating their overlapping motor control functions. These connections appear to be disrupted by alcohol intoxication, in turn associated with an explicit type of impaired driving behavior. PMID- 20028355 TI - Gene expression in the neuropeptide Y system during ethanol withdrawal kindling in rats. AB - BACKGROUND: Multiple episodes of ethanol intoxication and withdrawal result in progressive, irreversible intensification of the withdrawal reaction, a process termed "ethanol withdrawal kindling." Previous studies show that a single episode of chronic ethanol intoxication and withdrawal causes prominent changes in neuropeptide Y (NPY) and its receptors that have been implicated in regulating withdrawal hyperexcitability. This study for the first time examined the NPY system during ethanol withdrawal kindling. METHODS: Ethanol withdrawal kindling was studied in rats receiving 16 episodes of 2 days of chronic ethanol intoxication by intragastric intubations followed by 5 days withdrawal. The study included 6 groups: 4 multiple withdrawal episode (MW) groups [peak withdrawal plus (MW+)/minus (MW-) seizures, 3-day (MW3d), and 1-month (MW1mth) withdrawal], a single withdrawal episode group (SW), and an isocalorically fed control group. Gene expression of NPY and its receptors Y1, Y2, and Y5 was studied in the hippocampal dentate gyrus (DG) and CA3/CA1, as well as piriform cortex (PirCx), and neocortex (NeoCx). RESULTS: MW+/- as well as SW groups showed decreased NPY gene expression in all hippocampal areas compared with controls, but, in the DG and CA3, decreases were significantly smaller in the MW- group compared with the SW group. In the MW+/- and SW groups, Y1, Y2, and Y5 mRNA levels were decreased in most brain areas compared with controls; however, decreases in Y1 and Y5 mRNA were augmented in the MW+/- groups compared with the SW group. The MW+ group differed from the MW- group in the PirCx, where Y2 gene expression was significantly higher. CONCLUSION: Multiple withdrawal episodes reversibly decreased NPY and NPY receptor mRNA levels at peak withdrawal, with smaller decreases in NPY mRNA levels and augmented decreases in Y1/Y5 mRNA levels compared with a SW episode. Multiple withdrawal-induced seizures increased the Y2 mRNA levels in PirCx. These complex changes in NPY system gene expression could play a role in the ethanol withdrawal kindling process. PMID- 20028356 TI - The effect of moderate to heavy alcohol consumption on neuropsychological performance as measured by the repeatable battery for the assessment of neuropsychological status. AB - BACKGROUND: Excessive alcohol use is associated with damage to the structure and function of the brain and impairment of cognition and behavior. Traditional test batteries used to assess cognitive performance in alcoholics are extensive and costly, limiting their use across various clinical and research settings. The Repeatable Battery for the Assessment of Neuropsychological Status (RBANS) is a relatively new instrument that attempts to overcome some of these limitations. As yet the individual effect of moderate to heavy alcohol consumption on RBANS performance has not been examined. The primary aim of this study was to explore and quantify differences in performance between controls and drinkers on the RBANS and to examine the influence of age, gender, and alcohol use patterns on test performance. METHODS: Data from a subset of "Using Our Brains" (UoB) donors (n = 28) still actively drinking and meeting criteria for moderate to heavy alcohol use (30 to 80 g of ethanol per day) (Harper, 1988) and 28 matched controls (age, education, and premorbid Intelligence Quotient) were compared. RESULTS: Participants in the alcohol group performed below the healthy control group on the visuospatial and immediate memory index, and also on the RBANS total score p < 0.001 and showed a greater decline in RBANS scores from estimated cross sectional premorbid levels. There was a positive association between alcohol ingestion in the preceding 12 months and the language index p < 0.03 and the semantic fluency subtest (p < 0.03). Age was negatively associated with story memory (p < 0.02), coding (p < 0.001), list recognition (p < 0.01), story recall (p < 0.03), and figure recall (p < 0.02). CONCLUSION: Our results suggest that the RBANS is able to detect and characterize differences in verbal fluency, visuospatial skills, components of declarative memory, and psychomotor speed between healthy controls and moderate to heavy active alcohol users. Executive functions, commonly affected by alcoholism and not included in the RBANS, require assessment with additional measures. PMID- 20028357 TI - Betaine protects chronic alcohol and omega-3 PUFA-mediated down-regulations of PON1 gene, serum PON1 and homocysteine thiolactonase activities with restoration of liver GSH. AB - BACKGROUND: Paraoxonase (PON1) is an antioxidant enzyme that prevents LDL oxidation as well as detoxifies homocysteine thiolactone (HCTL), both of which can cause atherosclerosis. Chronic alcohol (ETOH) and high omega-3 polyunsaturated fatty acids (omega-3 PUFA) consumption may affect PON1 status presumably via reactive oxygen species by depleting liver glutathione (GSH), whereas betaine may counter their effects. Therefore, we investigated the influence of ETOH, omega-3 PUFA, and betaine on liver GSH, PON1 expression, lipid score, as well as serum PON1 and HCTLase activities. METHODS: Experimental rats belonging to various dietary groups were pair-fed with Lieber-DeCarli low (2.8% the dietary calories as omega3-fatty acids) and high (13.8% the dietary calories as omega3-fatty acids) menhaden fish alcohol-liquid diets with and without betaine (10 g/l diet) for 8 weeks after which liver PON1 mRNA, GSH, lipid score, and serum PON1, HCTLase, and ALT activities were measured. RESULTS: High omega-3 PUFA decreased liver PON1 mRNA expression, serum PON1, and HCTLase activity by 23% (p < 0.01), 20% (p < 0.05), and 28% (p < 0.05), respectively compared to the low omega-3 PUFA group. ETOH decreased PON1 mRNA expression by 25 and 30% (p < 0.01) with concomitant 27% (p < 0.05) and 38% (p < 0.01), decrease in liver GSH levels in low and high omega-3 PUFA groups, respectively. Correspondingly, serum PON1 activity decreased by 23% (p < 0.05) and 58% (p < 0.01) while serum HCTLase activity decreased by 25% (p < 0.05) and 59% (p < 0.01) in the low and high omega 3 PUFA ETOH groups, respectively. Betaine restored liver PON1 mRNA expressions in low and high omega-3 PUFA ETOH groups with parallel restorations of PON1 activity and liver GSH. Concomitantly, betaine reduced hepatosteatosis accompanied by alleviation of liver injury caused by chronic alcohol and high omega-3 PUFA. CONCLUSIONS: Based on these results, we conclude that dietary betaine not only atheroprotective by restoring liver GSH that quenches free radicals, but also may alleviate liver injury by reducing hepatosteatosis. PMID- 20028359 TI - Subjective response to alcohol: a critical review of the literature. AB - BACKGROUND: Subjective response to alcohol (SR), which reflects individual differences in sensitivity to the pharmacological effects of alcohol, may be an important endophenotype in understanding genetic influences on drinking behavior and alcohol use disorders (AUDs). SR predicts alcohol use and problems and has been found to differ by a range of established risk factors for the development of AUDs (e.g., family history of alcoholism). The exact pattern of SR associated with increased risk for alcohol problems, however, remains unclear. The Low Level of Response Model (LLR) suggests that high-risk individuals experience decreased sensitivity to the full range of alcohol effects, while the Differentiator Model (DM) asserts that high risks status is associated with increased sensitivity to alcohol's positive effects but decreased sensitivity to negative effects. AIMS: The current paper (1) reviews two prominent models of subjective response, (2) reviews extant laboratory-based research on subjective response, (3) highlights remaining gaps in our understanding and assessment of subjective response, and (4) encourages collaborative efforts to address these methodological and conceptual concerns. METHODS: This paper reviews studies which employed placebo controlled and non-placebo-controlled alcohol challenge paradigms to assess a range of alcohol effects including impairment, stimulation, and sedation. RESULTS: The research literature provides at least partial support for both the LLR and DM models. High-risk individuals have been shown to have a reduced response to alcohol with respect to sedative or impairing effects, particularly on the descending limb of the blood alcohol curve (BAC). There is also evidence that ascending limb stimulant effects are more pronounced or operate differently for high-risk individuals. DISCUSSION: Despite commendable advances in SR research, important questions remain unanswered. Inconsistent results across studies may be attributable to a combination of an inadequate understanding of the underlying construct and methodological differences across studies (e.g., number and timing of assessments across the BAC, inclusion of a placebo condition). With respect to the underlying construct, existing measures fail to adequately distinguish between cognitive/behavioral impairment and sedation, aspects of which may be perceived positively (e.g., anxiolysis) due to their ability to act as negative reinforcers. CONCLUSIONS: Addressing the concerns raised by the current review will be integral to making meaningful scientific progress in the field of subjective response. PMID- 20028358 TI - Ethanol-induced oxidative stress and mitochondrial dysfunction in rat placenta: relevance to pregnancy loss. AB - BACKGROUND: Ethanol consumption during pregnancy increases the risk of early pregnancy loss and causes intrauterine growth restriction. We previously showed that chronic gestational exposure to ethanol impairs placentation, and that this effect is associated with inhibition of insulin and insulin growth factor signaling. Since ethanol also causes oxidative stress and DNA damage, we extended our investigations to assess the role of these pathological processes on placentation and placental gene expression. METHODS: Pregnant Long Evans rats were pair-fed liquid diets containing 0% or 24% ethanol by caloric content. Placentas harvested on gestation day 16 were used to examine DNA damage, lipid peroxidation, apoptosis, mitochondrial gene/protein and hormonal gene expression in relation to ethanol exposure. RESULTS: Gestational exposure to ethanol increased fetal resorption, and trophoblast apoptosis/necrosis, oxidative stress, DNA damage, and lipid peroxidation. These adverse effects of ethanol were associated with increased expression of pro-apoptotic (Bax and Bak) and reduced levels of the anti-apoptotic Bcl-2 protein. In addition, increased trophoblast apoptosis proneness was associated with p53-independent activation of p21, reduced mitochondrial gene and protein expression, and dysregulated expression of prolactin (PRL) family hormones that are required for implantation and pregnancy related adaptations. CONCLUSIONS: Chronic gestational exposure to ethanol increases fetal demise due to impaired survival and mitochondrial function, increased oxidative stress, DNA damage and lipid peroxidation, and dysregulated expression of prolactin family hormones in placental trophoblasts. PMID- 20028360 TI - Mechanisms of postural control in alcoholic men and women: biomechanical analysis of musculoskeletal coordination during quiet standing. AB - BACKGROUND: Excessive sway during quiet standing is a common sequela of chronic alcoholism even with prolonged sobriety. Whether alcoholic men and women who have remained abstinent from alcohol for weeks to months differ from each other in the degree of residual postural instability and biomechanical control mechanisms has not been directly tested. METHOD: We used a force platform to characterize center of-pressure biomechanical features of postural sway, with and without stabilizing conditions from touch, vision, and stance, in 34 alcoholic men, 15 alcoholic women, 22 control men, and 29 control women. Groups were matched in age (49.4 years), general intelligence, socioeconomic status, and handedness. Each alcoholic group was sober for an average of 75 days. RESULTS: Analysis of postural sway when using all 3 stabilizing conditions versus none revealed diagnosis and sex differences in ability to balance. Alcoholics had significantly longer sway paths, especially in the anterior-posterior direction, than controls when maintaining erect posture without balance aids. With stabilizing conditions the sway paths of all groups shortened significantly, especially those of alcoholic men, who demonstrated a 3.1-fold improvement in sway path difference between the easiest and most challenging conditions; the remaining 3 groups, each showed a approximately 2.4-fold improvement. Application of a mechanical model to partition sway paths into open-loop and closed-loop postural control systems revealed that the sway paths of the alcoholic men but not alcoholic women were characterized by greater short-term (open-loop) diffusion coefficients without aids, often associated with muscle stiffening response. With stabilizing factors, all 4 groups showed similar long-term (closed loop) postural control. Correlations between cognitive abilities and closed-loop sway indices were more robust in alcoholic men than alcoholic women. CONCLUSIONS: Reduction in sway and closed-loop activity during quiet standing with stabilizing factors shows some differential expression in men and women with histories of alcohol dependence. Nonetheless, enduring deficits in postural instability of both alcoholic men and alcoholic women suggest persisting liability for falling. PMID- 20028361 TI - Ecological associations of alcohol outlets with underage and young adult injuries. AB - OBJECTIVE: This paper argues that associations between rates of 3 specific problems related to alcohol (i.e., accidents, traffic crashes, and assaults) should be differentially related to densities of alcohol outlets among underage youth and young adults based upon age-related patterns of alcohol outlet use. METHODS: Zip code-level population models assessed local and distal effects of alcohol outlets upon rates of hospital discharges for these outcomes. RESULTS: Densities of off-premise alcohol outlets were significantly related to injuries from accidents, assaults, and traffic crashes for both underage youth and young adults. Densities of bars were associated with more assaults and densities of restaurants were associated with more traffic crash injuries for young adults. CONCLUSIONS: The distribution of alcohol-related injuries relative to alcohol outlets reflect patterns of alcohol outlet use. PMID- 20028362 TI - Differential effects of ethanol on serum GABAergic 3alpha,5alpha/3alpha,5beta neuroactive steroids in mice, rats, cynomolgus monkeys, and humans. AB - BACKGROUND: Acute ethanol administration increases plasma and brain levels of progesterone and deoxycorticosterone-derived neuroactive steroids (3alpha,5alpha) 3-hydroxypregnan-20-one (3alpha,5alpha-THP) and (3alpha,5alpha)-3,21 dihydroxypregnan-20-one (3alpha,5alpha-THDOC) in rats. However, little is known about ethanol effects on GABAergic neuroactive steroids in mice, nonhuman primates, or humans. We investigated the effects of ethanol on plasma levels of 3alpha,5alpha- and 3alpha,5beta-reduced GABAergic neuroactive steroids derived from progesterone, deoxycorticosterone, dehydroepiandrosterone, and testosterone using gas chromatography-mass spectrometry. METHODS: Serum levels of GABAergic neuroactive steroids and pregnenolone were measured in male rats, C57BL/6J and DBA/2J mice, cynomolgus monkeys, and humans following ethanol administration. Rats and mice were injected with ethanol (0.8 to 2.0 g/kg), cynomolgus monkeys received ethanol (1.5 g/kg) intragastrically, and healthy men consumed a beverage containing 0.8 g/kg ethanol. Steroids were measured after 60 minutes in all species and also after 120 minutes in monkeys and humans. RESULTS: Ethanol administration to rats increased levels of 3alpha,5alpha-THP, 3alpha,5alpha THDOC, and pregnenolone at the doses of 1.5 g/kg (+228, +134, and +860%, respectively, p < 0.001) and 2.0 g/kg (+399, +174, and +1125%, respectively, p < 0.001), but not at the dose of 0.8 g/kg. Ethanol did not alter levels of the other neuroactive steroids. In contrast, C57BL/6J mice exhibited a 27% decrease in serum 3alpha,5alpha-THP levels (p < 0.01), while DBA/2J mice showed no significant effect of ethanol, although both mouse strains exhibited substantial increases in precursor steroids. Ethanol did not alter any of the neuroactive steroids in cynomolgus monkeys at doses comparable to those studied in rats. Finally, no effect of ethanol (0.8 g/kg) was observed in men. CONCLUSIONS: These studies show clear species differences among rats, mice, and cynomolgus monkeys in the effects of ethanol administration on circulating neuroactive steroids. Rats are unique in their pronounced elevation of GABAergic neuroactive steroids, while this effect was not observed in mice or cynomolgus monkeys at comparable ethanol doses. PMID- 20028364 TI - Intoxication with bourbon versus vodka: effects on hangover, sleep, and next-day neurocognitive performance in young adults. AB - BACKGROUND: This study assessed the effects of heavy drinking with high or low congener beverages on next-day neurocognitive performance, and the extent to which these effects were mediated by alcohol-related sleep disturbance or alcoholic beverage congeners, and correlated with the intensity of hangover. METHODS: Healthy heavy drinkers age 21 to 33 (n = 95) participated in 2 drinking nights after an acclimatization night. They drank to a mean of 0.11 g% breath alcohol concentration on vodka or bourbon one night with matched placebo the other night, randomized for type and order. Polysomnography recordings were made overnight; self-report and neurocognitive measures were assessed the next morning. RESULTS: After alcohol, people had more hangover and more decrements in tests requiring both sustained attention and speed. Hangover correlated with poorer performance on these measures. Alcohol decreased sleep efficiency and rapid eye movement sleep, and increased wake time and next-day sleepiness. Alcohol effects on sleep correlated with hangover but did not mediate the effects on performance. No effect of beverage congeners was found except on hangover severity, with people feeling worse after bourbon. Virtually no sex differences appeared. CONCLUSIONS: As drinking to this level affects complex cognitive abilities, safety could be affected, with implications for driving and for safety sensitive occupations. Congener content affects only how people feel the next day so does not increase risk. The sleep disrupting effects of alcohol did not account for the impaired performance so other mechanisms of effect need to be sought. As hangover symptoms correlate with impaired performance, these might be contributing to the impairment. PMID- 20028363 TI - Common genetic contributions to alcohol and cannabis use and dependence symptomatology. AB - BACKGROUND: Despite mounting evidence that use of and dependence on alcohol and cannabis are influenced by heritable factors, the extent to which heritable influences on these phenotypes overlap across the 2 substances has only rarely been explored. In the current study, we quantified cross-substance overlap in sources of variance and estimated the degree to which within-substance associations between use and dependence measures are attributable to common genetic and environmental factors for alcohol and cannabis. METHODS: The sample was comprised of 6,257 individuals (2,761 complete twin pairs and 735 singletons) from the Australian Twin Registry, aged 24 to 36 years. Alcohol and cannabis use histories were collected via telephone diagnostic interviews and used to derive an alcohol consumption factor, a frequency measure for cannabis use, and DSM-IV alcohol and cannabis dependence symptom counts. Standard genetic analyses were conducted to produce a quadrivariate model that provided estimates of overlap in genetic and environmental influences across the 4 phenotypes. RESULTS: Over 60% of variance in alcohol consumption, cannabis use, and cannabis dependence symptoms, and just under 50% of variance in alcohol dependence (AD) symptoms were attributable to genetic sources. Shared environmental factors did not contribute significantly to the 4 phenotypes. Nearly complete overlap in heritable influences was observed for within-substance measures of use and dependence symptoms. Genetic correlations across substances were 0.68 and 0.62 for use and dependence symptoms, respectively. CONCLUSIONS: Common heritable influences were evident for alcohol and cannabis use and for AD and cannabis dependence symptomatology, but findings indicate that substance-specific influences account for the majority of the genetic variance in the cannabis use and dependence phenotypes. By contrast, the substantial correlations between alcohol use and AD symptoms and between cannabis use and cannabis dependence symptoms suggest that measures of heaviness of use capture much of the same genetic liability to alcohol- and cannabis-related problems as dependence symptomatology. PMID- 20028365 TI - Exercise neuroprotection in a rat model of binge alcohol consumption. AB - BACKGROUND: Excessive alcohol intake produces structural and functional deficits in corticolimbic pathways that are thought to underlie cognitive deficits in the alcohol use disorders (AUDs). Animal models of binge alcohol administration support the direct link of high levels of alcohol consumption and neurotoxicity in the hippocampus and surrounding cortex. In contrast, voluntary wheel running enhances hippocampal neurogenesis and generally promotes the health of neurons. METHODS: We investigated whether voluntary exercise prior to binge alcohol exposure could protect against alcohol-induced cell loss. Female Long-Evans rats exercised voluntarily for 14 days before undergoing 4 days of binge alcohol consumption. Brains were harvested immediately after the last dose of alcohol and examined for various histological markers of neurodegeneration, including both cell death (FluoroJade B) and cell birth (Ki67) markers. RESULTS: Rats that exercised prior to binge exposure were significantly less behaviorally intoxicated, which was not a result of enhanced hepatic metabolism. Rats that exercised prior to binge alcohol consumption had reduced loss of dentate gyrus granule cells and fewer FluoroJade B positive cells in the dentate gyrus and associated entorhinal-perirhinal cortex compared to nonexercisers. However, exercise did not protect against cell death in the piriform cortex nor protect against alcohol-induced decreases in cell proliferation, evidenced by a similar alcohol-induced reduction in Ki67 labeled cells between exercise and sedentary rats. CONCLUSIONS: We conclude that exercise can reduce behavioral sensitivity to ethanol intoxication and protect vulnerable brain areas from alcohol-induced cell death. Exercise neuroprotection of alcohol-induced brain damage has important implications in understanding the neurobiology of the AUDs as well as in developing novel treatment strategies. PMID- 20028366 TI - Exploring the relationship between depressive and anxiety symptoms and neuronal response to alcohol cues. AB - BACKGROUND: Depressive and anxiety symptoms tend to co-occur with heavy drinking. Specifically, their presence may exacerbate the severity and intractability of heavy drinking. Similarly, heavy drinking may increase the risk for and experience of depressive and anxiety symptoms. Although depressive and anxiety symptoms have been significantly correlated with alcohol craving in cue-exposure paradigms, physiological responses have not always mapped onto emotional responses. Therefore, this study sought to examine the role of depressive and anxiety symptoms using a more basic science approach, through examining functional brain changes. METHODS: Seventy nontreatment seeking, heavy drinking adults were recruited through a college campus (n = 45 men; mean age = 22.8). They completed measures of drinking, smoking, depressive symptoms, anxiety symptoms, and a functional magnetic resonance imaging (fMRI) cue-exposure paradigm. RESULTS: As hypothesized, depressive symptoms were positively correlated with activation during the alcohol (vs. appetitive control) cue in the insula, cingulate, ventral tegmentum, striatum, and thalamus (cluster-corrected p < 0.05, z = 2.3). Similarly, anxiety symptoms were positively correlated with activation during the alcohol (vs. appetitive control) cue in the striatum, thalamus, insula, and inferior frontal, mid-frontal, and cingulate gyri (cluster corrected p < 0.05, z = 2.3). CONCLUSIONS: Significant correlations were found between depressive symptoms, anxiety symptoms, and differential brain activation in response to an alcohol versus an appetitive control cue in an fMRI paradigm. Moreover, the pattern of activation mapped onto expected regions. This study strongly supports the posited relationships between depressive symptoms, anxiety symptoms, and differential brain activation in an alcohol cue-exposure paradigm with a sample of heavy drinking adults. PMID- 20028367 TI - Chronic alcohol consumption disrupted cholesterol homeostasis in rats: down regulation of low-density lipoprotein receptor and enhancement of cholesterol biosynthesis pathway in the liver. AB - BACKGROUND: Chronic alcohol consumption causes alcoholic liver disease, which is associated, or initiated, with dysregulated lipid metabolism. Very recent evidence suggested that dysregulated cholesterol metabolism plays an important role in the pathogenesis of alcoholic fatty liver diseases, however, the effects of chronic alcohol exposure on cholesterol homeostasis have not been well studied and underlying mechanisms behind are still elusive. METHODS: Male Sprague-Dawley rats weighing 250 +/- 5.5 g (mean +/- SEM) divided into 2 groups (8 rats per group) and pair-fed with liquid diets containing (in percent of energy intake) 18% protein, 35% fat, 12% carbohydrate, and 35% either ethanol (ethanol diet) or an isocaloric maltose-dextrin mixture (control diet), according to Lieber and De Carli, for 4 weeks. RESULTS: Long-term excessive alcohol feeding to rats caused fatty liver and liver injury, which was associated with disrupted cholesterol homeostasis, characterized by increased hepatic cholesterol levels and hypercholesterolemia. Hepatic cholesterol increases were concomitant with constantly activated sterol regulatory element-binding protein-2 (SREBP-2) in the liver and increased expression of 3-hydroxy-3-methyl-glutaryl-CoA (HMG-CoA) reductase, a rate-limiting enzyme for cholesterol de novo synthesis, indicating enhanced cholesterol biosynthesis. Alcohol-induced hypercholesterolemia was accompanied by decreased LDL receptor (LDLr) levels in the liver. Further investigations revealed that chronic alcohol exposure increased hepatic proprotein convertase subtilisin/kexin type 9 (PCSK9) contents to down-regulate LDLr via a post-translational mechanism. Moreover, alcohol feeding suppressed extracellular signal-regulated kinase (ERK) activation in the liver. In vitro studies showed that inhibition of ERK activation was associated with decreased LDLr expression in HepG2 cells. CONCLUSIONS: Our study provides the first evidence that both increased PCSK9 expression and suppressed ERK activation in the liver contributes to alcohol-induced hypercholesterolemia in rats. PMID- 20028369 TI - Selective ovine serum albumin allergy in an adult. PMID- 20028368 TI - Neuropeptide Y signaling in the central nucleus of amygdala regulates alcohol drinking and anxiety-like behaviors of alcohol-preferring rats. AB - BACKGROUND: The neuropeptide Y (NPY) system of the central nucleus of amygdala (CeA) has been shown to be involved in anxiety and alcoholism. In this study, we investigated the molecular mechanisms by which NPY in the CeA regulates anxiety and alcohol drinking behaviors using alcohol-preferring (P) rats as an animal model. METHODS: Alcohol-preferring rats were bilaterally cannulated targeting the CeA and infused with artificial cerebrospinal fluid (aCSF) or NPY. Alcohol drinking and anxiety-like behaviors were assessed by the 2-bottle free-choice paradigm and light/dark box (LDB) exploration test, respectively. The levels of NPY and related signaling proteins were determined by the gold immunolabeling procedure. The mRNA levels of NPY were measured by in situ RT-PCR. Double immunofluorescence labeling was performed to observe the colocalization of NPY and Ca(2+)/calmodulin-dependent protein kinase IV (CaMK IV). RESULTS: We found that NPY infusion into the CeA produced anxiolytic effects, as measured by the LDB exploration test, and also decreased alcohol intake in P rats. NPY infusion into the CeA significantly increased levels of CaMK IV and phosphorylated cAMP responsive element-binding (pCREB) protein and increased mRNA and protein levels of NPY, but produced no changes in protein levels of CREB or the catalytic alpha subunit of protein kinase A (PKA-Calpha) in the CeA. We also observed that alcohol intake produced anxiolytic effects in P rats in the LDB test and also increased NPY expression and protein levels of pCREB and PKA-Calpha without modulating protein levels of CREB or CaMK IV, in both the CeA and medial nucleus of amygdala. In addition, we found that CaMK IV-positive cells were co-localized with NPY in amygdaloid structures of P rats. CONCLUSIONS: These results suggest that NPY infusion may increase the expression of endogenous NPY in the CeA, which is most likely attributable to an increase in CaMK IV-dependent CREB phosphorylation and this molecular mechanism may be involved in regulating anxiety and alcohol drinking behaviors of P rats. PMID- 20028370 TI - Tracheobronchomalacia in wheezing young children poorly responsive to asthma therapy. PMID- 20028371 TI - Analysis of the high affinity IgE receptor genes reveals epistatic effects of FCER1A variants on eczema risk. AB - BACKGROUND: High levels of total and allergen-specific IgE levels are a key feature in allergic diseases. The high-affinity receptor for IgE, which is composed of one alpha (FCER1A), one beta (FCER1B), and two gamma (FCER1G) subunits, represents the central receptor of IgE-induced reactions. In a genome wide association scan, we recently identified associations between functional FCER1A variants and total serum IgE levels. Previous studies had reported linkage and association of FCER1B variants with IgE and atopic traits. The FCER1G gene has not yet been investigated with regard to atopy. Filaggrin (FLG) is the strongest known risk gene for eczema, in particular the allergic subtype of eczema. METHODS: We investigated the association of FCER1A, FCER1B, and FCER1G variants with IgE in a large population-based cohort (n = 4261) and tested for epistatic effects using the model-based multifactor dimensionality reduction (MB MDR) method. In addition, we investigated a potential interaction between FLG and FCER1A variants in a large collection of eczema cases (n = 1018) and population controls. RESULTS: Three strongly correlated FCER1A polymorphisms were significantly associated with total and specific IgE levels as well as allergic sensitization. No associations were seen for FCER1B and FCER1G. After adjustment for FLG effects, a significant epistatic effect of the FCER1A variants rs10489854 and rs2511211 on eczema risk was detected. CONCLUSIONS: These results suggest that FCER1A variants by themselves and in combination influence IgE levels and act synergistically to influence eczema risk. PMID- 20028372 TI - Specific immunotherapy suppresses Th2 responses via modulating TIM1/TIM4 interaction on dendritic cells. AB - BACKGROUND: Specific immunotherapy (SIT) is the only curable remedy for allergic disorders currently; however, the underlying mechanism is not fully understood yet. This study aimed to elucidate the mechanism of SIT on suppressing TIM4 (T cell immunoglobulin mucin domain molecule 4) expression in dendritic cells (DCs) and modulating the skewed T helper 2 (Th2) responses in patients with airway allergy. METHODS: Twenty patients with allergic rhinitis (AR) were treated with SIT for 3 months. Before and after SIT, the expression of TIM4 in peripheral DC and TIM1 in Th2 cells was examined. The role of Fc gamma receptor (FcgammaR) I and II in modulating the expression of TIM4 in DCs was investigated. RESULTS: The interaction of TIM1/TIM4 played a critical role in sustaining the polarization status of Th2 cells in AR patients. Cross-linking FcgammaRI by antigen/IgG complexes increased the production of TIM4 by dendritic cells via upregulating tumor necrosis factor-alpha in DCs. Exposure to microbial products promoted the expression of FcgammaRI in DCs that further increased the expression of TIM4. Exposure to specific antigens alone upregulated the expression of FcgammaRII in DCs, that suppressed the expression of TIM4. CONCLUSIONS: We conclude that SIT suppresses the skewed Th2 responses via disrupting the interaction of TIM1/TIM4 in antigen-specific Th2 cells. PMID- 20028373 TI - Gaps in anaphylaxis management at the level of physicians, patients, and the community: a systematic review of the literature. AB - Diagnosis and management of anaphylaxis can be a challenge because reactions are often unexpected and progress quickly. The focus of anaphylaxis management has mostly been on the acute episode, with little attention given to the long-term management of patients at risk. This is compounded by conflicting information in current guidelines and a general lack of agreement among clinicians about which management strategies are the most appropriate. We systematically reviewed the literature to identify and summarize studies that investigated gaps in anaphylaxis management. Our search included MEDLINE, EMBASE, CINAHL, and Evidence Based Medicine Reviews. Studies were included if they addressed an outcome describing gaps in anaphylaxis knowledge, education, anaphylaxis management, and quality of life (QOL). Populations of interest were health care professionals involved in the care of patients at risk for anaphylaxis, and patients of any age, their parents, caregivers, and teachers in primary care, hospital or community settings. Of 5014 citations that were identified, the final 59 studies (selected from 75 full-text articles) met the inclusion criteria. Two hundred and two gaps were identified and classified according to major themes: gaps in knowledge and anaphylaxis management (physicians and patients); gaps in follow-up care (physicians); and QOL of patients and caregivers. Findings from this systematic review revealed gaps in anaphylaxis management at the level of physicians, patients, and the community. Findings will be used to provide a basis for developing interventional strategies to help address these deficiencies. PMID- 20028374 TI - Towards evidence-based medicine in specific grass pollen immunotherapy. AB - When initiating grass pollen immunotherapy for seasonal allergic rhinoconjunctivitis, specialist physicians in many European countries must choose between modalities of differing pharmaceutical and regulatory status. We applied an evidence-based medicine (EBM) approach to commercially available subcutaneous and sublingual Gramineae grass pollen immunotherapies (SCIT and SLIT) by evaluating study design, populations, pollen seasons, treatment doses and durations, efficacy, quality of life, safety and compliance. After searching MEDLINE, Embase and the Cochrane Library up until January 2009, we identified 33 randomized, double-blind, placebo-controlled trials (including seven paediatric trials) with a total of 440 specific immunotherapy (SIT)-treated subjects in seven trials (0 paediatric) for SCIT with natural pollen extracts, 168 in three trials (0 paediatric) for SCIT with allergoids, 906 in 16 trials (five paediatric) for natural extract SLIT drops, 41 in two trials (one paediatric) for allergoid SLIT tablets and 1605 in five trials (two paediatric) for natural extract SLIT tablets. Trial design and quality varied significantly within and between SIT modalities. The multinational, rigorous trials of natural extract SLIT tablets correspond to a high level of evidence in adult and paediatric populations. The limited amount of published data on allergoids prevented us from judging the level of evidence for this modality. PMID- 20028375 TI - X-chromosome Forkhead Box P3 polymorphisms associate with atopy in girls in three Dutch birth cohorts. AB - BACKGROUND: The Forkhead Box P3 (FOXP3) gene, located on the X-chromosome, encodes a transcription factor that directs T cells toward a regulatory phenotype. Regulatory T cells may suppress development of atopy. We evaluated whether single-nucleotide polymorphisms (SNPs) of FOXP3 are associated with atopy development in childhood. METHODS: Seven SNPs in FOXP3 were genotyped in 3062 children (51% boys) participating in the Allergenic study, which consists of three Dutch birth cohorts (PIAMA, PREVASC and KOALA). Association of FOXP3 SNPs with total serum IgE and sensitisation (presence of specific serum IgE to egg, milk, and indoor, i.e. house-dust mite, cat, and/or dog allergens) was investigated at ages 1, 2, 4, and 8. Analysis of variance and logistic regression were performed, stratified for gender. RESULTS: Our most consistent finding was observed for sensitisation to egg and indoor allergens. In girls, five FOXP3 SNPs (rs5906761, rs2294021, rs2294019, rs6609857 and rs3761548) were significantly associated with sensitisation to egg at ages 1 and 2 and with sensitisation to indoor allergens at age 2 (P < 0.05), but not at 4 and 8, a finding that was observed across the three cohorts. Rs5906761 and rs2294021 were associated with remission of sensitisation to food allergens in boys, as tested in the PIAMA cohort. CONCLUSION: This is the first study showing across three cohorts that X chromosomal FOXP3 genotypes may contribute to development of sensitisation against egg and indoor allergens in girls in early childhood. In addition, an association with remission of sensitisation to food allergens existed in boys only. PMID- 20028377 TI - Cranial muscles of the Sulawesi Babirusa (Babyrousa celebensis). AB - The detailed muscular anatomy of the head of the Sulawesi Babirusa (Babyrousa celebensis) is described for the first time. The results show that the muscular anatomy of the Babyrousa is very similar to the pig genus Sus, despite long geological separation from it. Some differences were noted: the M. parietoauricularis was more clearly separated into two parts in the Babirusa than in the domestic pig; the rostral fibres of the M. levator anguli oculi reached the infraorbital sinus in the Babirusa but only as far as the medial corner of the eye in the domestic pig; the M. palatinus of the Babirusa is paired and did not reach the Os palatinum, unlike its description in the domestic pig and the Platysma pars zygomatica originates from the fascia of the neck in the Babirusa, whereas that of the domestic pig originates largely or entirely from the scapula. PMID- 20028378 TI - Immunohistochemistry of pancreatic development in cattle and pig. AB - The aim of this study was to characterize bovine and porcine pancreatic development by immunohistochemistry. In the pig, staining for both glucagon and insulin was noted at day 19. In cattle, glucagon staining was observed at day 25 and insulin staining from day 26. In both species, glucagon-stained cells were abundant initially, but later insulin-stained cells became most abundant. A few cells displayed co-localization of glucagon and insulin staining during initial development in both species. Initially, most of the cells of the pancreatic primordia and the duodenal epithelium displayed Pdx-1-staining. All insulin stained cells displayed Pdx-1-stained nuclei, whereas no glucagon-stained cells did so. Many Pdx-1-stained cells lacked insulin staining, but with development, the relative number of these cells diminished. Nkx6.1-staining was initially seen in a pattern similar to that for Pdx-1, but was lacking duodenal staining. Subsequently, the number of Nkx6.1-stained cells diminished, but increased again to a level where practically all insulin-stained cells also presented Nkx6.1 staining. Glucagon-stained cells, on the other hand, never had Nkx6.1 staining. In conclusion, the localization of the two transcription factors, Pdx-1 and Nkx6.1, demonstrated that pancreas development appears to be controlled by mechanisms comparable with those operating in humans. PMID- 20028380 TI - JTE-607, a multiple cytokine production inhibitor, induces apoptosis accompanied by an increase in p21waf1/cip1 in acute myelogenous leukemia cells. AB - Proinflammatory cytokines and growth factors have been thought to play crucial roles in the pathology of acute myelogenous leukemia (AML) by supporting the proliferation and survival of AML cells in an autocrine and paracrine manner, although further elucidation is required. JTE-607 was originally identified as a multiple cytokine inhibitor that suppresses production of proinflammatory cytokines from lipopolysaccharide (LPS)-stimulated peripheral blood mononuclear cells. Herein, we report that JTE-607 exhibits inhibitory activity on the growth of AML cell lines accompanying reduction of the proinflammatory cytokine and growth factor production. In in vitro studies, JTE-607 suppressed expression and production of cytokines, which are spontaneously up-regulated in AML cell lines. JTE-607 also abrogated proliferation of AML cells in a concentration range in which colony formation of normal bone marrow cells was not affected. The growth inhibition by JTE-607 was characterized by induction of cell-cycle arrest at the S-phase and apoptosis, accompanied by a decrease in c-Myc and increase in p21(waf1/cip1). In a leukemia model engrafted with U-937 cells, JTE-607 significantly prolonged survival in mice and reduced human cytokine mRNA levels in the bone marrow. These results suggest the usefulness of JTE-607 in therapeutic applications for patients with hypercytokinemia and aggressive AML cell proliferation. PMID- 20028379 TI - QTL for body weight, morphometric traits and stress response in European sea bass Dicentrarchus labrax. AB - Natural mating and mass spawning in the European sea bass (Dicentrarchus labrax L., Moronidae, Teleostei) complicate genetic studies and the implementation of selective breeding schemes. We utilized a two-step experimental design for detecting QTL in mass-spawning species: 2122 offspring from natural mating between 57 parents (22 males, 34 females and one missing) phenotyped for body weight, eight morphometric traits and cortisol levels, had been previously assigned to parents based on genotypes of 31 DNA microsatellite markers. Five large full-sib families (five sires and two dams) were selected from the offspring (570 animals), which were genotyped with 67 additional markers. A new genetic map was compiled, specific to our population, but based on the previously published map. QTL mapping was performed with two methods: half-sib regression analysis (paternal and maternal) and variance component analysis accounting for all family relationships. Two significant QTL were found for body weight on linkage group 4 and 6, six significant QTL for morphometric traits on linkage groups 1B, 4, 6, 7, 15 and 23 and three suggestive QTL for stress response on linkage groups 3, 14 and 23. The QTL explained between 8% and 38% of phenotypic variance. The results are the first step towards identifying genes involved in economically important traits like body weight and stress response in European sea bass. PMID- 20028376 TI - International variations in associations of allergic markers and diseases in children: ISAAC Phase Two. AB - BACKGROUND: Circulating allergen-specific IgE (sIgE) and skin prick tests (SPT) are used to define atopy. Downregulation of local inflammatory responsiveness has been proposed to explain a low prevalence of positive SPTs in less affluent countries. We analysed the association between SPTs, total and allergen-specific IgE and their relationships to allergic symptoms in centres with diverse living conditions. METHODS: Cross-sectional studies of stratified random samples of 8 to 12-year-old children (n = 7461) used the standardized methodology of Phase Two of the International Study of Asthma and Allergies in Childhood (ISAAC). Symptoms of asthma, rhinitis and eczema were ascertained by parental questionnaires. Skin examination, hypertonic saline bronchial challenge, six aeroallergen SPTs and measurements of serum total IgE and sIgE were performed. RESULTS: In nonaffluent countries, a higher proportion of children with positive SPT had no detectable sIgE (range 37-61%) than in affluent countries (0-37%). Total serum IgE was associated with all disease outcomes among children with both positive SPT and sIgE (P < 0.001), but only with self-reported eczema in children with negative SPTs and negative sIgE. CONCLUSIONS: The international pattern of discordance between SPT and sIgE results did not support the downregulation hypothesis. Among children with no evidence of sensitization to common aeroallergens, increased total IgE contributes little to the risk of wheeze and rhinitis in the general population but may play a role in eczema. PMID- 20028381 TI - Clinical implications for nestin protein expression in breast cancer. AB - Recently, it was observed that nestin is preferentially expressed in basal/myoepithelial cells of the mammary gland, and that this intermediate filament may be used as a myoepithelial marker. However, the clinical and prognostic implications of nestin as a marker for breast cancer are still unclear. We examined mastectomy specimens from 150 breast cancers and matching, adjacent non-cancerous tissues using immunohistochemistry and western blotting. Overall, triple-negative breast cancers - that is, breast cancers that do not express estrogen receptors (ER), progesterone receptors (PR), or human epidermal growth factor receptor 2 (HER2/neu) - had higher expression rates for nestin than the other breast cancers (57.14%vs 9.30%; P < 0.001). In triple-negative breast cancers, significantly increased nestin expression rates were observed in patients with lymph node metastasis compared with those without node metastasis (25.00%vs 76.92%; P = 0.032). A similar phenomenon was observed for invasive ductal carcinomas compared with ductal carcinoma in situ (16.67%vs 73.33%; P = 0.046). Nestin expression was also found to be significantly related to ER, PR, and P53 expression (P < 0.05). Nestin expression was associated with both shorter breast cancer-specific survival and poor relapse-free survival in the lymph node positive group (P = 0.028 and P = 0.012 respectively). After Cox regression was carried out, nestin was not shown to be an independent prognostic factor for breast cancer. These findings substantiate the possibility of using nestin as a marker for triple-negative breast cancer. Triple-negative breast cancer progression is associated with nestin; however, the underlying mechanisms of this relationship require further investigation. PMID- 20028382 TI - Resveratrol induces apoptosis and cell cycle arrest of human T24 bladder cancer cells in vitro and inhibits tumor growth in vivo. AB - Resveratrol, a naturally occurring polyphenolic antioxidant compound present in grapes and red wine, has been reported to hold various biochemical responses. In this preliminary study, we evaluate the chemopreventive potential of resveratrol against bladder cancer and its mechanism of action. Treatment of bladder cancer cells with resveratrol resulted in a significant decrease in cell viability. Resveratrol induced apoptosis through the modulation of Bcl-2 family proteins and activation of caspase 9 and caspase 3 followed by poly(ADP-ribose) polymerase degradation. Treatment with resveratrol led to G(1) phase cell cycle arrest in T24 cells by activation of p21 and downregulation of cyclin D1, cyclin-dependent kinase 4, and phosphorylated Rb. Resveratrol also inhibited the phosphorylation of Akt, whereas the phosphorylation of p38 MAPK was enhanced. In addition, resveratrol treatment decreased the expression of vascular endothelial growth factor and fibroblast growth factor-2, which might contribute to the inhibition of tumor growth on the bladder cancer xenograft model. These findings suggest that reveratrol could be an important chemoprevention agent for bladder cancer. PMID- 20028383 TI - Phase I study of irinotecan and doxifluridine for metastatic colorectal cancer focusing on the UGT1A1*28 polymorphism. AB - Although individuals carrying the UGT1A1 allele *28 have an increased risk of severe toxicities associated with irinotecan, no phase I study has been conducted based on the polymorphism. This report presents the recommended doses of irinotecan for patients with the respective genotypes. Twenty-seven patients with advanced colorectal cancer were enrolled in this study, and the UGT1A1*28 polymorphism was genotyped before chemotherapy. One course of chemotherapy consisted of irinotecan infused once every 2 weeks at 70, 100, 120, and 150 mg/m(2) at dose levels 1, 2, 3, and 4, respectively, and doxifluridine was administered orally. This treatment continued for at least 12 weeks. The dose limiting toxicity was determined as grade 3 hematological and non-hematological toxicities for the TA(6)/TA(6) (6/6) and TA(6)/TA(7) (6/7) genotypes. The pharmacokinetics of irinotecan, SN-38, and SN-38 glucuronide, was assessed at dose level 2. Eighteen and nine patients had the 6/6 and 6/7 genotypes, respectively. The maximum tolerated dose (MTD) was not observed up to dose level 4 in patients with the 6/6 genotype. In contrast, MTD was observed at dose level 2 (100 mg/m(2)) in patients with the 6/7 genotype. Patients with the 6/7 genotype had a significantly higher area under the plasma time-concentration curve (0 infinity) SN-38 (P = 0.022) and biliary index (P = 0.030) than those with 6/6. The recommended starting doses of biweekly irinotecan for phase II/III were 150 mg/m(2) for patients with the UGT1A1 6/6 genotype and 70 mg/m(2) for those with the 6/7 genotype, respectively. The gene polymorphism should be considered when determining the precise recommended doses to be administered in phase I studies. PMID- 20028384 TI - Irrespective of CD34 expression, lineage-committed cell fraction reconstitutes and re-establishes transformed Philadelphia chromosome-positive leukemia in NOD/SCID/IL-2Rgammac-/- mice. AB - Stem cells of acute myeloid leukemia (AML) have been identified as immunodeficient mouse-repopulating cells with a Lin(-)CD34(+)38(-) phenotype similar to normal hematopoietic stem cells. To identify the leukemia-propagating stem cell fraction of Philadelphia chromosome-positive (Ph(+)) leukemia, we serially transplanted human leukemia cells from patients with chronic myeloid leukemia blast crisis (n = 3) or Ph(+) acute lymphoblastic leukemia (n = 3) into NOD/SCID/IL-2Rgammac(-/-) mice. Engrafted cells were almost identical to the original leukemia cells as to phenotypes, IGH rearrangements, and karyotypes. CD34(+)CD38(-)CD19(+), CD34(+)38(+)CD19(+), and CD34(-)CD38(+)CD19(+) fractions could self-renew and transfer the leukemia, whereas the CD34(-)CD38(+)CD19(+) fraction did not stably propagate in NOD/SCID mice. These findings suggest that leukemia-repopulating cells in transformed Ph(+) leukemia are included in a lineage-committed but multilayered fraction, and that CD34(+) leukemia cells potentially emerge from CD34(-) populations. PMID- 20028385 TI - Hydroxyurea upregulates NKG2D ligand expression in myeloid leukemia cells synergistically with valproic acid and potentially enhances susceptibility of leukemic cells to natural killer cell-mediated cytolysis. AB - Valproic acid (VPA), a histone deacetylase inhibitor, upregulates NKG2D ligands (NKG2DLs) on some monocytic and lymphoid leukemic cells. However, its effect on myeloid leukemia cells and synergistic agents that can augment the effect of VPA remains unknown. Of the various myeloid cell lines examined, OUN-1, a chronic myelogenous leukemia cell line, showed the most prominent upregulation of MICA/B and ULBP2 in response to VPA. The NKG2DL upregulation was observed only in leukemic cells without apoptosis and the effect was abrogated by pretreatment of cells with caffeine, an inhibitor of ATM/ATR. Several activators of ATM/ATR were screened for their effect on NKG2DL expression, but only hydroxyurea (HU) efficiently upregulated both MICA/B and ULPB2 expression on the cell line. VPA and HU synergistically upregulated the NKG2DLs on OUN-1 cells as well as primary leukemic cells from some patients with acute myeloid leukemia. The upregulation of NKG2DLs by VPA and/or HU was associated with increased transcription of each NKG2DL gene. OUN-1 cells treated with VPA + HU were more susceptible to killing by natural killer (NK) cells than untreated cells and the enhanced cytotoxicity of NK cells was blocked by the treatment of NK cells with anti-NKG2D monoclonal antibodies. The same concentrations of VPA and HU did not affect the cytotoxicity of NK cells against OUN-1 cells. These data suggest that VPA and HU might enhance the NK cell-mediated antileukemia effect by increasing the susceptibility of myeloid leukemic cells to NK cells. PMID- 20028386 TI - Rubratoxin A specifically and potently inhibits protein phosphatase 2A and suppresses cancer metastasis. AB - Although cytostatin analog protein phosphatase 2A (PP2A)-specific inhibitors are promising candidates of a new type of anticancer drug, their development has been hindered because of their liability. To find new classes of PP2A-specific inhibitors, we conducted a screening with microbial metabolites and found that rubratoxin A, a classical mycotoxin, is a highly specific and potent inhibitor of the enzyme. While rubratoxin A inhibits PP2A at Ki = 28.7 nm, it hardly inhibited any other phosphatases examined. Rubratoxin B, a close analog, also specifically but weakly inhibits PP2A at Ki = 3.1 microM. The inhibition of intracellular PP2A in cultured cells is obviously observed with 20 microM rubratoxin A treatment for 3 h, inducing the overphosphorylation in PP2A substrate proteins. Although rubratoxins and cytostatin differ in the apparent structures, these compounds share similarities in the structures in detail and PP2A-binding manners. Rubratoxin A showed higher suppression of tumor metastasis and reduction of the primary tumor volume than cytostatin in mouse experiments. As a successor of cytostatin analogs, rubratoxin A should be a good compound leading to the development of antitumor drugs targeting PP2A. PMID- 20028387 TI - Role of tumor-associated lymphatic endothelial cells in metastasis: a study of epithelial ovarian tumor in vitro. AB - Tumor-associated lymphatic endothelial cells (TLEC) could play a key role in the process of tumor metastasis. The aim of this study was to investigate the effect of TLECs that were isolated from human epithelial ovarian tumor (EOT) on ovarian cancer cell line CAOV-3 in vitro. First, TLECs in EOT were detected by immunochemistry and flow cytometry, then marked by lymphatic endothelial cell (LEC) marker LYVE-1, isolated by magnetic beads, and cultured in vitro. The cells were identified by immunostaining of LEC markers LYVE-1, Prox-1, Podoplanin, VEGFR-3, and pan-endothelial cell marker CD31. TLECs from EOT can be detected, cultured, and identified in vitro successfully. The effects of TLECs on invasion and migration of CAOV-3 cells were investigated by 12-well Boyden chamber; the proliferation effect was studied by counting the Trypan blue exclusion cell number. Furthermore, changes in MMP-2/9 secreted by CAOV-3 cells treated with TLEC were shown using real-time PCR and zymography, and TIMP-1/2 was detected by real-time PCR. In vitro, TLECs can enhance invasion and migration of CAOV-3 cells, but have no significant effect on proliferation. It was clear that the expression of MMP-9 increased and TIMP-2 decreased in CAOV-3 cells treated by TLECs, and the increasing of MMP-9 was confirmed by zymography. TLECs from EOT can enhance migration and invasion of human ovarian carcinoma cell line in vitro, and the possible mechanism was through activation of MMP-9/TIMP-2. PMID- 20028388 TI - CD44+ slow-cycling tumor cell expansion is triggered by cooperative actions of Wnt and prostaglandin E2 in gastric tumorigenesis. AB - Similar to normal tissue stem cells, cancer stem cells (CSCs) are thought to be quiescent or slow-cycling and, thereby, insensitive to chemo- and radiotherapies. CD44, a cell surface component that interacts with the extracellular matrix, has been found to be highly expressed in CSCs of several solid tumors. However, the relevancy between CD44(+) cells and slow-cycling cells and the underlying mechanisms for the emergence of CD44(+) CSCs during tumorigenesis have not been elucidated. Here we show that a gastric gland residing at the squamo-columnar junction (SCJ) in normal mouse stomach contains CD44(+) stem cell-like slow cycling cells and that this characteristic CD44(+) gland was expanded by prostaglandin E2 (PGE(2)) and Wnt signaling in K19-Wnt1/C2mE mouse, a genetic mouse model for gastric tumorigenesis. The analysis of three transgenic mouse lines, K19-Wnt1, K19-C2mE and K19-Wnt1/C2mE, revealed that the expansion of CD44(+) SCJ cells is triggered by PGE(2)-mediated signaling and is prominently enhanced by the addition of Wnt activation. Furthermore, each expanded CD44(+) gland in gastric tumor of K19-Wnt1/C2mE mouse contains a few BrdU label-retaining quiescent or slow-cycling cells, suggesting that the CD44(+) SCJ cells in normal mouse are candidates for the cell-of-origin of gastric CSCs. These observations suggest that PGE(2)-mediated inflammatory signaling and Wnt signaling cooperatively trigger the expansion of CD44(+) slow-cycling stem-like cells in SCJ, leading to development of lethal gastric tumors in mice. PMID- 20028389 TI - Altered phenotype of lymphatic endothelial cells induced by highly metastatic OTSCC cells contributed to the lymphatic metastasis of OTSCC cells. AB - The lymphatic endothelial cell (LEC) is an interactive surface for cancer cells. This article aims to explore cancer cell-induced changes of LEC, and study the tumor-lymphatic endothelium interaction. Here, LECs were co-cultured with highly and poorly metastatic tongue cancer cells. The differences in biologic behaviors and gene expression profiles between them were examined. The results showed that LECs induced by highly metastatic cancer cells displayed abnormal biologic behaviors, and could secrete chemokines to promote the migration of cancer cells. Therefore, biologic properties and functional status of LECs in oral tongue squamous cell carcinoma (OTSCC) might be a positive factor in lymphatic dissemination. PMID- 20028390 TI - Reduction and recombination of fingerprints of different design increase compound recall and the structural diversity of hits. AB - We report an advanced 'hybrid fingerprint' design concept specifically for the purpose of scaffold hopping. The generation of hybrid fingerprints includes two major steps. In the 'fingerprint reduction' step, bit positions of different types of fingerprints (e.g. substructural and pharmacophore fingerprints) are ranked according to their statistical significance and ability to discriminate between specifically active compounds and database decoys. On the basis of bit ranking, subsets containing the most discriminatory bit positions are determined. In the subsequent 'fingerprint recombination' step, bit subsets from different fingerprints are combined to yield a new compound class-directed fingerprint representation for similarity searching. Here, we generate hybrids from multiple fingerprints and analyze their search performance in comparison with parental fingerprints on compound activity classes that exclusively consist of molecules with unique core structures and that exhibit different levels of intra-class structural diversity. Fingerprint reduction is found to be a critical component of hybrid design. The resulting compound class-directed hybrid fingerprints further increase the similarity search performance and scaffold hopping potential of their parental fingerprints. Thus, fingerprint reduction and recombination improve compound recall and increase the structural diversity of hits. PMID- 20028391 TI - The aromatic amino acid residues of tumor necrosis factor receptor-1-derived peptide are important for promoting differentiation of neural stem cells. AB - Neural stem cells have the self-renewal capacity and the ability to differentiate into all types of nerve cells. We previously reported that the tumor necrosis factor receptor-1-derived peptide promotes neural differentiation of fetal rat hippocampal neural stem cells. The tumor necrosis factor receptor-1-derived peptide contains six aromatic amino acid residues among its 14 amino acid residues. To clarify the role of these aromatic amino acid residues in the action of tumor necrosis factor receptor-1-derived peptide on neural stem cells, we synthesized mutant peptides, in which aromatic residues were substituted with alanine, and we assessed their effects. Substitution of the tyrosine residue at position 103 (Y(103)) or 106 (Y(106)), the tryptophan residue at position 107 (W(107)), or the phenylalanine residue at position 112 (F(112)) or 115 (F(115)), decreased the ability of the peptide to promote neurite outgrowth of neural stem cells depending on their concentration. These data suggest that although all five aromatic amino acid residues mediate the action of the tumor necrosis factor receptor-1-derived peptide, their order of importance in this activity is F(115) > Y(103) > W(107) > Y(106) and F(112). PMID- 20028392 TI - Structural findings of 2-phenylindole-3-carbaldehyde derivatives for antimitotic activity by FA-sMLR QSAR analysis. AB - Antimitotic agents prevent the mitosis process of cell cycle and are generally used for the treatment of cancer with good clinical success. QSAR analysis was performed on 33 reported 2-phenylindole-3-carbaldehyde derivatives to find out the structural requirements of these compounds for higher antimitotic activity. The dataset was divided into test set and training set by k-MCA. Factor analysis stepwise multiple linear regression (FA-sMLR) method was used to develop statistically significant QSAR equations. QSAR analysis showed importance of topological indices like RTSA indices, structural information content indices, Balaban-type index from van der Waals weighted distance matrix Jhetv, rotatable bond fraction RBF, and Lovasz-Pelikan index LP1 on biological activity. The study also shows significance of electrostatic potential charges of different atoms as well as principal moment of inertia along Y axis and number of nitrogen atoms toward antimitotic activity of these compounds. PMID- 20028393 TI - 3D-QSAR studies of arylcarboxamides with inhibitory activity on InhA using pharmacophore-based alignment. AB - Enoyl acyl carrier protein reductase (InhA) is a promising target for the development of antituberculosis drugs. The InhA-bound conformation of an indole-5 amide inhibitor (Genz 10850) (PDB code: IP44) was used to build a pharmacophore model by LigandScout. This model was then successfully used to identify the bioactive conformation and align 40 structurally diverse arylcarboxamide derivatives. Comparative molecular field analysis (CoMFA) and comparative molecular similarity indices analysis (CoMSIA) were performed on arylcarboxamides based InhA inhibitors based on pharmacophore alignment. The best prediction was obtained with CoMSIA model combining steric and electrostatic fields (, r(2) = 0.972). The model was validated by an external test set, which gave a good predictive value (). Graphical interpretation of the results revealed important structural features of the zarylcarboxamides related to the active site of InhA. The results may be exploited for further design and virtual screening for some novel InhA inhibitors. PMID- 20028394 TI - In vitro studies on the antioxidant and protective effect of 2-substituted -8 hydroxyquinoline derivatives against H(2)O(2)-induced oxidative stress in BMSCs. AB - Novel 2-vinyl-8-hydroxyquinoline derivatives as potential antioxidants and regulators of H(2)O(2)-induced oxidative stress in rat bone marrow mesenchymal stem cells (MSCs) are first reported. The antiradical properties and the reducing power of these compounds were assessed using 2, 2-diphenyl-1-picrylhydrazyl (DPPH) and auto-oxidation of pyrogallol method, respectively. The activity against lipid peroxidation was determined using ammonium thiocyanate method. The results revealed that introduction of electron-donating groups at 2nd position decreased the antioxidant activities of 8-hydroxyquinoline derivatives. In addition, compound 4, the structure of which is similar to melatonin, exhibited superior antioxidant activities in scavenging DPPH free radical, O(2) free radical, and anti-LPO activities. Except for compounds 7, 12, and 15, the other compounds exhibited a stimulatory effect on MSCs growth. Using hydrogen peroxide (H(2)O(2)), we also investigated the protective efficacy of 2-vinyl-8 hydroxyquinoline derivatives against oxidative stress-induced cell death of MSCs. Cell viability assayed by MTT method indicated that exposure of MSCs cultures to hydrogen peroxide resulted in a concentration-dependent decrease in cell viability, and compounds 4 and 5 at given concentration (2.62 x 10(-3) m) could protect MSCs against H(2)O(2)-induced oxidative stress in bone mesenchymal stem cell (BMSCs). PMID- 20028395 TI - Synthesis and antileishmanial activity of lipidic amino alcohols. AB - In this work, a number of lipidic amino alcohols wereas synthesized and evaluated in vitro on cultures of Leishmania amazonensis and Leishmania chagasi. Nine amino alcohols showed inhibition of L. chagasi growth, and seven of them showed inhibition of L. amazonensis with IC(50) below 10 microm. Compound 11f was more active than the reference drug amphotericin B against L. chagasi promastigote forms. PMID- 20028396 TI - How much binding affinity can be gained by filling a cavity? AB - Binding affinity optimization is critical during drug development. Here, we evaluate the thermodynamic consequences of filling a binding cavity with functionalities of increasing van der Waals radii (-H, -F, -Cl, and CH(3)) that improve the geometric fit without participating in hydrogen bonding or other specific interactions. We observe a binding affinity increase of two orders of magnitude. There appears to be three phases in the process. The first phase is associated with the formation of stable van der Waals interactions. This phase is characterized by a gain in binding enthalpy and a loss in binding entropy, attributed to a loss of conformational degrees of freedom. For the specific case presented in this article, the enthalpy gain amounts to -1.5 kcal/mol while the entropic losses amount to +0.9 kcal/mol resulting in a net 3.5-fold affinity gain. The second phase is characterized by simultaneous enthalpic and entropic gains. This phase improves the binding affinity 25-fold. The third phase represents the collapse of the trend and is triggered by the introduction of chemical functionalities larger than the binding cavity itself [CH(CH(3))(2)]. It is characterized by large enthalpy and affinity losses. The thermodynamic signatures associated with each phase provide guidelines for lead optimization. PMID- 20028397 TI - Synthesis, preferred conformation, and membrane activity of medium-length peptaibiotics: tylopeptin B. AB - The solid-phase synthesis and full chemical characterization of the medium-length (14-amino acid residues) peptaibol with antibiotic properties of tylopeptin B, originally extracted from the fruiting body of the mushroom Tylopilus neofelleus, are described. These data are accompanied by the results on the solution-phase synthesis via the segment condensation approach of a selected, side-chain protected, analog. A solution conformational analysis, performed by the combined use of FTIR absorption, circular dichroism, and 2D-NMR (the latter technique coupled to molecular dynamics calculations), favors the conclusion that the 3D structure of tylopeptin B is largely helical with a preference for the alpha- or the 3(10)-helix type depending upon the nature of the solvent. Helix topology and (partial) amphiphilic character are responsible for the observed membrane modifying properties of this peptaibiotic. PMID- 20028399 TI - Folding studies of pH-dependent collagen peptides. AB - Synthetic collagen model peptides containing carboxylate-modified hydroxyproline residues (P(E)) have been shown to form a collagen triple helix at low pH (1). Based on the fact that P(E)-containing peptides unfold on demand by altering the pH, we investigated the folding kinetics of a series of these collagen-based peptides. Refolding data indicated that the introduction of P(E) residues within a peptide affects the stability and refolding of the collagen peptides through electrostatic interactions and steric hindrance. Moreover, the specific placement of the P(E) residues within the collagen peptides allowed for selective testing for possible nucleation sites at the termini or the center of the peptides. Specifically, the data demonstrated that collagen peptides may nucleate from either terminally or internally located repeating sequences of Pro-Hyp-Gly. These studies indicate that the P(E) residue may play a useful role as a tool for studying the folding mechanism of synthetic collagen peptides. PMID- 20028398 TI - N-methylated cyclic enkephalin analogues retain high opioid receptor binding affinity. AB - In an effort to improve the bioavailability of the non-selective, cyclic enkephalin analogues H-Dmt-c[d-Cys-Gly-Phe-d(or L)-Cys]NH(2) (Dmt = 2',6' dimethyltyrosine), analogues N-methylated at the Phe(4) and/or Cys(5) residue were synthesized. In comparison with the non-methylated parent peptides, all mono and N-di-methylated analogues in general retained high binding affinities at all three opioid receptors and high opioid agonist potencies in functional opioid activity assays. The results indicate that the progressive conformational restriction in these compounds upon mono- and di-N-methylation did not significantly affect the in vitro opioid activity profile. A low-energy conformer identified for the conformationally most restricted analogue of the series, H-Dmt c[D-Cys-Gly-Phe(NMe)-L-Cys(NMe)]NH(2) (6), showed good spatial overlap of the essential pharmacophoric moieties with those in the proposed mu receptor-bound conformation of the mu-selective opioid peptide JOM-6 [H-Tyr-c(S-Et-S)[D-Cys-Phe D-Pen]NH(2)] (Pen = penicillamine) [Mosberg M.I. and Fowler C.B. (2002) J Peptide Res; 60:329-335], in agreement with the moderate mu selectivity determined for this compound. An analogue of 6 containing (2S)-2-methyl-3-(2,6-dimethyl-4 hydroxyphenyl)propanoic acid [(2S)-Mdp] in place of Dmt(1) was an opioid antagonist with quite high opioid receptor binding affinities and can be expected to show improved bioavailability because of its further increased lipophilicity and reduced hydrogen-bonding capacity. PMID- 20028400 TI - High throughput receptor-based virtual screening under ZINC database, synthesis, and biological evaluation of ketol-acid reductoisomerase inhibitors. AB - Ketol-acid reductoisomerase (KARI; EC 1.1.1.86) catalyzes the second common step in branched-chain amino acid biosynthesis. This enzyme is an important target for drug design. Based on the crystal structure of ketol-acid reductoisomerase/N hydroxy-N-isopropyloxamate (IpOHA) complex, we have carried out high throughput receptor-based virtual screening of the ZINC/drug like database (2 000 000 compounds) to look for novel inhibitors of KARI for the first time. Some novel compounds were found to inhibit rice KARI in vitro among 15 procured compounds. This method can provide useful information for further design and discovery of KARI inhibitors. PMID- 20028401 TI - AP24163 inhibits the gatekeeper mutant of BCR-ABL and suppresses in vitro resistance. AB - Mutation in the ABL kinase domain is the principal mechanism of imatinib resistance in patients with chronic myelogenous leukaemia. The second generation BCR/ABL inhibitors nilotinib and dasatinib effectively inhibit most imatinib resistance variants, but are ineffective against the gatekeeper mutant, T315I. Gatekeeper mutation activates the kinase by stabilizing the hydrophobic spine. Here, we describe that the rationally designed compound AP24163 can inhibit native and gatekeeper mutants of the BCR/ABL kinase. Structural modelling suggests that AP24163 affects the flexibility of the P-loop and destabilizes the active conformation by disrupting the hydrophobic spine. In vitro screening for drug resistance identified clones with compound mutations involving both the P loop and T315I. Our studies provide structural insights for the design of inhibitors against the gatekeeper mutant and suggest that up-front combination therapy may be required to prevent the emergence of compound-resistant mutations. PMID- 20028402 TI - Dome-shaped tumour with surface changes on the abdominal wall. PMID- 20028404 TI - Career choices in dermatology. AB - Reforms to both undergraduate and postgraduate training have resulted in limited exposure to clinical dermatology, and therefore we need to target advice and clinical attachments to the most appropriate stages of training to ensure that dermatology is considered as a career option by medical students. A questionnaire survey was undertaken to determine when and how doctors chose their career in dermatology. An electronic questionnaire was sent to 709 dermatologists (response rate 44%). Of the respondents, 49% considered dermatology after medical school attachment and 61% were influenced by the jobs they had done as a pre registration house officer (PRHO) or senior house officer (SHO). Careers advice was lacking, with only 11% receiving it while at medical school and 29% during their PRHO/SHO years. Contact with an inspiring clinician was the most influential factor in choosing dermatology. Both medical students and junior doctors, should have opportunities to learn from and work with clinical dermatologists. PMID- 20028405 TI - Tender red scalp and spreading rash. PMID- 20028406 TI - Crusts and vacciniform scars on sun-exposed skin. PMID- 20028407 TI - Chronic recurrent follicular papules and plaques on the face. PMID- 20028408 TI - Erythematous macules in a 14-year-old girl. PMID- 20028409 TI - Neonatal urticaria due to American ginseng or not? PMID- 20028410 TI - An alternative treatment option for compensatory hyperhidrosis after endoscopic thoracic sympathectomy. PMID- 20028412 TI - Neglect of genetic diversity in implementation of the convention on biological diversity. PMID- 20028413 TI - Biodiversity conservation in agricultural landscapes: challenges and opportunities of coffee agroforests in the Western Ghats, India. AB - The new approaches advocated by the conservation community to integrate conservation and livelihood development now explicitly address landscape mosaics composed of agricultural and forested land rather than only protected areas and largely intact forests. We refer specifically to a call by Harvey et al. (2008) to develop a new approach based on six strategies to integrate biodiversity conservation with sustainable livelihoods in Mesoamerican landscape mosaics. We examined the applicability of this proposal to the coffee agroforests of the Western Ghats, India. Of the six strategies, only one directly addresses livelihood conditions. Their approach has a clear emphasis on conservation and, as currently formulated risks repeating the failures of past integrated conservation and development projects. It fails to place the aspirations of farmers at the core of the agenda. Thus, although we acknowledge and share the broad vision and many of the ideas proposed by this approach, we urge more balanced priority setting by emphasizing people as much as biodiversity through a careful consideration of local livelihood needs and aspirations. PMID- 20028414 TI - Testing hypotheses of bird extinctions at Rio Palenque, Ecuador, with informal species lists. AB - Informally gathered species lists are a potential source of data for conservation biology, but most remain unused because of questions of reliability and statistical issues. We applied two alternative analytical methods (contingency tests and occupancy modeling) to a 35-year data set (1973-2007) to test hypotheses about local bird extinction. We compiled data from bird lists collected by expert amateurs and professional scientists in a 2-km(2) fragment of lowland tropical forest in coastal Ecuador. We tested the effects of the following on local extinction: trophic level, sociality, foraging specialization, light tolerance, geographical range area, and biogeographic source. First we assessed extinction on the basis of the number of years in which a species was not detected on the site and used contingency tests with each factor to compare the frequency of expected and observed extinction events among different species categories. Then we defined four multiyear periods that reflected different stages of deforestation and isolation of the study site and used occupancy modeling to test extinction hypotheses singly and in combination. Both types of analyses supported the biogeographic source hypothesis and the species-range hypothesis as causes of extinction; however, occupancy modeling indicated the model incorporating all factors except foraging specialization best fit the data. PMID- 20028415 TI - The power of bioenergy-related standards to protect biodiversity. AB - The sustainable production of bioenergy is vital to avoiding negative impacts on environmental goods such as climate, soil, water, and especially biodiversity. We propose three key issues that should be addressed in any biodiversity risk mitigation strategy: conservation of areas of significant biodiversity value; mitigation of negative effects related to indirect land-use change; and promotion of agricultural practices with few negative impacts on biodiversity. Focusing on biodiversity concerns, we compared principles and criteria set to address biodiversity and other environmental and social issues in seven standards (defined here as commodity-based standards or roundtables, or relevant European legislation): five voluntary initiatives related to bioenergy feedstocks, the Renewable Transport Fuel Obligation (United Kingdom), and the European Renewable Energy Source Directive. Conservation of areas of significant biodiversity value was fairly well covered by these standards. Nevertheless, mitigation of negative impacts related to indirect land-use change was underrepresented. Although the EU directive, with its bonus system for the use of degraded land and a subquota system for noncrop biofuels, offered the most robust standards to mitigate potential negative effects, all of the standards fell short in promoting agricultural practices with low negative impacts on biodiversity. We strongly recommend that each standard be benchmarked against related standards, as we have done here, and that efforts should be made to strengthen the elements that are weak or missing. This would be a significant step toward achieving a bioenergy industry that safeguards Earth's living heritage. PMID- 20028416 TI - Synergistic action of the novel HSP90 inhibitor NVP-AUY922 with histone deacetylase inhibitors, melphalan, or doxorubicin in multiple myeloma. AB - Heat shock protein 90 (HSP90) is a promising target for tumor therapy. The novel HSP90 inhibitor NVP-AUY922 has preclinical activity in multiple myeloma, however, little is known about effective combination partners to design clinical studies. Multiple myeloma cell lines, OPM-2, RPMI-8226, U-266, LP-1, MM1.S, and primary myeloma cells were exposed to NVP-AUY922 and one of the combination partners histone deacetylase inhibitor NVP-LBH589, suberoylanilide hydroxamic acid (SAHA), melphalan, or doxorubicin, either simultaneously or in sequential patterns. Effects on cell proliferation and apoptosis were determined. Synergistic effects were evaluated using the method of Chou and Talalay. Combined sequential incubation with NVP-AUY922 and SAHA showed that best synergistic effects were achieved with 24 h preincubation with SAHA followed by another 48 h of combination treatment. Combination of NVP-AUY922 with SAHA, NVP-LBH589, melphalan, or doxorubicin resulted in synergistic inhibition of viability, with strong synergy (combination index < 0.3) in the case of melphalan. Importantly, resistance of the RPMI-8226 cell line and relative resistance of some primary myeloma cells against NVP-AUY922 could be overcome by combination treatment. These data show impressive synergistic action of the novel HSP90 inhibitor NVP AUY922 with melphalan, doxorubicin, NVP-LBH589, and SAHA in multiple myeloma and build the frame work for clinical trials. PMID- 20028417 TI - Extramedullary haematopoiesis in lymph node in a patient of T cell leukemia. PMID- 20028419 TI - Comparison of kaolin and tissue factor activated thromboelastography in haemophilia. AB - A limitation of bypassing agent therapy for haemophilia patients with inhibitors is the absence of a laboratory assay, which predicts the clinical response to treatment. Recent investigations have demonstrated the potential for thromboelastography to assess the effects of bypassing agent therapy in this patient population. While tissue factor activation has been used in several prior studies, a recent multicentre study failed to demonstrate an expected concentration-response effect of rFVIIa and called into question the tissue factor activation methods that have been employed. A comparison of kaolin to two concentrations of tissue factor as the activation method for thromboelastography was investigated in patients with haemophilia. We performed kaolin and tissue factor activated thromboelastography on blood from inhibitor and non-inhibitor patients with and without addition of rFVIIa and rFVIII. The results demonstrate that kaolin leads to a longer R, K and angle than the higher dilution of tissue factor (1:17 000) at baseline (no factor) and after addition of rFVIIa for both the inhibitor and non-inhibitor patients. Kaolin led to a longer R and K in comparison to a low dilution of tissue factor (1:42 000) following the addition of rFVIIa in the inhibitor patients. The longer R and K allows for better discrimination of the effects of rFVIIa thus making kaolin the most sensitive activation method in this setting. Thus kaolin activated thromboelastography should be considered an effective, perhaps the most effective, activator when utilizing thromboelastography to assess the effects of rFVIIa in haemophilia patients with inhibitors. PMID- 20028420 TI - Reply to 'Differential response to bypassing agents complicates treatment in patients with haemophilia and inhibitors' (Haemophilia 2009; 15: 3-10). PMID- 20028421 TI - Assessing the benefits of FEIBA prophylaxis in haemophilia patients with inhibitors. AB - SUMMARY: Prophylaxis with concentrates of factor VIII has become the standard of care for patients with severe haemophilia A because of its ability to prevent joint and other bleeding events. Recent evidence suggests that the prophylactic use of bypassing therapy--activated prothrombin complex concentrate (aPCC) and recombinant activated factor VII (rFVIIa)--provides similar benefits to haemophilia patients with inhibitors. To assess the efficacy and safety of prophylaxis with the aPCC FEIBA, a meta-analysis was conducted of six studies and a total of 34 inhibitor patients. The mean prophylactic dose was 78.5 U kg(-1), typically infused three to four times weekly. A total of 31 of 33 patients (94%) for whom bleeding data were available prior to prophylaxis experienced a decrease in the rate of haemorrhage, albeit minor in some patients, and, regardless of the type of haemorrhage measured, had on average a 63.9% reduction in bleeding episodes during FEIBA prophylaxis. In the three studies that specifically assessed all joint bleeding, the 18 patients evaluated experienced an average reduction in annual joint bleeding of 74% while on prophylactic regimens. Among the four patients in this group who received concurrent immune tolerance induction and the 14 patients treated with prophylaxis only, annual joint bleeding decreased by an average of 79% and 78%, respectively. Anamnesis following FEIBA exposure was observed in some patients but had no impact on prophylactic efficacy. No thrombotic or other complications were reported. These results suggest that the prophylactic administration of FEIBA can be an effective and safe method for reducing bleeding events in patients with haemophilia A and inhibitors. PMID- 20028422 TI - Ten novel factor VIII (F8C) mutations in eighteen haemophilia A families detected in Singapore. PMID- 20028423 TI - Adenotonsillectomy in patients with desmopressin responsive mild bleeding disorders: a review of the literature. AB - SUMMARY: Many patients with mild inherited bleeding disorders such as von Willebrand disease (VWD), mild haemophilia A (HA) and platelet function defects (PFD) undergo adenoidectomy and/or tonsillectomy (AT) procedures each year. Management of bleeding in these patients can be challenging, as little published data exist to guide haemostatic management during these relatively common procedures. Therefore, the literature was reviewed to identify AT procedures in patients with 1-deamino-8-D-argine vasopressin responsive mild bleeding disorders. The review revealed no randomized prospective trials of haemostatic management in this patient population. Case reports and small case series identified 144 patients who had AT procedures. Frequency of desmopressin and antifibrinolytic dosing varied widely. Fifteen percentage of patients experienced postoperative bleeding with nearly half being early (<24 h) bleeding and half being late (>24 h) bleeding. Hyponatraemia complicated the procedures in 47% of cases and six hyponatremic seizures were reported. Issues identified by this review that need to be addressed in future clinical trials include type and amount of fluid restriction when utilizing desmopressin, duration of antifibrinolytic therapy and duration and frequency of desmopressin dosing. PMID- 20028424 TI - Cost-effectiveness analysis of different imaging strategies for diagnosis of haemophilic arthropathy. AB - SUMMARY: Physiotherapy and radiography of the joints are standard diagnostic strategies for assessment of haemophilic arthropathy. The use of ultrasonography as an adjunct tool for early diagnosis of haemophilic arthropathy may optimize factor replacement therapy. The objective of this study was to compare costs and effectiveness of physiotherapy, radiography and ultrasonography (intervention strategy, IS) with physiotherapy and radiography alone (standard care strategy, SCS) for diagnosing soft tissue and osteocartilaginous changes in haemophilic joints. We retrospectively compared costs and effectiveness of IS vs. SCS in knees, ankles and elbows of 31 children (age range, 4-17 years) with haemophilia A (n = 30) or B (n = 1) (IS, n = 11; SS, N = 20). Direct health care costs were measured from the provincial health care perspective. Effectiveness was measured by false-negative (FN) rates in each study arm by comparing presence or absence of abnormalities of physiotherapy and imaging exams to the reference standard measure (MRI). In scenario 1, all diagnostic tests matched with MRI. In scenario 2, at least one diagnostic test matched with MRI. The IS was more costly [incremental cost/100 patients, Canadian (CND) $4987] and more effective (incremental effectiveness, FNs/100 patients for scenario 1, -4.09, and for scenario 2, -41) for both scenarios. The incremental cost-effectiveness ratios for scenario 1 and for scenario 2 were CND$1166 and CDN$116 per FN result averted per 100 patients, respectively. In conclusion, in the short-term, the incorporation of ultrasonography in a test set for diagnosis of haemophilic arthropathy substantially improved the diagnostic performance of this test set, however at an increased cost. PMID- 20028425 TI - The optimal mode of delivery for the haemophilia carrier expecting an affected infant is caesarean delivery. AB - While a majority of affected infants of haemophilia carriers who deliver vaginally do not suffer a head bleed, the outcome of labour cannot be predicted. A planned vaginal delivery puts a woman at risk of an abnormal labour and operative vaginal delivery, both of which predispose to intracranial haemorrhage. Furthermore, vaginal delivery does not eliminate the risk to the haemophilia carrier herself. Overall, maternal morbidity and mortality from planned vaginal delivery are not significantly different from those from planned caesarean delivery. Caesarean delivery is recommended or elected now in conditions other than haemophilia carriage, where the potential benefits are not nearly as great. Additionally, vaginal delivery of the haemophilia carrier poses medical/legal risks if the infant is born with cephalohaematoma or intracranial haemorrhage. Caesarean delivery allows for a planned, controlled delivery. Caesarean delivery reduces the risk of intracranial haemorrhage by an estimated 85% and the risk can be nearly eliminated by performing elective caesarean delivery before labour. Therefore, after a discussion of the maternal and foetal risks with planned vaginal delivery versus planned caesarean delivery, haemophilia carriers should be offered the option of an elective caesarean delivery. PMID- 20028426 TI - Management of patients with factor V deficiency: open issues from the challenging history of a woman with anaphylactic transfusion reactions. PMID- 20028427 TI - Efficacy and safety of pegylated interferon alpha-2a therapy for chronic hepatitis C in HIV-infected patients with haemophilia. AB - The objective of this study was to evaluate the efficacy and safety of pegylated interferon (PEG-IFN) alpha-2a monotherapy in a cohort of Chinese haemophilic patients co-infected with human immunodeficiency virus (HIV)/hepatitis C virus (HCV) and undergoing highly active antiretroviral drugs therapy. Twenty-two (n = 22) patients with CD4 lymphocyte counts over 200 cells microL(-1) were treated with 180 microg of PEG-IFN alpha-2a subcutaneously once in a week for 48 weeks. HCV load (HCV RNA), HIV load (HIV RNA) and CD4 lymphocyte counts were measured at baseline and 4, 12, 24, 48 and 72 weeks after initiation of anti-HCV therapy. Efficacy and safety were analysed according to baseline CD4 status (> or =350 cells microL(-1)). Significant HCV-RNA decreases (>1 log(10) copies mL(-1)) were observed through week 72 after PEG-INF alpha-2a monotherapy across both CD4 strata. CD4 status was not associated with treatment outcomes as evaluated using rapid viral response rate (P = 0.655), early viral response rate (P = 0.387), end of-treatment viral response rate (P = 1.000) or sustained viral response rate (SVR, P = 0.674). A sustained virological response was achieved in nine patients (41%), five with genotype 2a (83%) and four with genotype 1b (25%, P = 0.023). SVR was HCV genotype dependent. Eleven patients required a dose reduction in PEG IFN alpha-2a. PEG-IFN alpha-2a monotherapy could be considered as a safe and effective option for the treatment of HCV infection in HIV patients with haemophilia, particularly in resource-limited settings. While higher CD4 lymphocyte counts resulted in greater HCV-RNA reduction, HCV genotype was a predictor for sustained virological response. PMID- 20028428 TI - The extensive polymorphism of KIR genes. AB - The functions of human natural killer (NK) cells are controlled by diverse families of antigen receptors. Prominent among these are the killer cell immunoglobulin-like receptors (KIR), a family of genes clustered in one of the most variable regions of the human genome. Within this review we discuss the vast polymorphism of the KIR gene complex which rivals that of the human leucocyte antigen (HLA) complex. There are several aspects to this polymorphism. Initially there is presence/absence of individual KIR genes, with four of these genes, termed framework genes, being present in all individuals tested to date, except on those very occasional instances when the gene has been deleted. Within each gene, alleles are present at different frequencies. We provide details of a new website that enables convenient searching for data on KIR gene, allele and genotype frequencies in different populations and show how these frequencies vary in different worldwide populations and the high probability of individuals differing in their KIR repertoire when both gene and allele polymorphism is considered. The KIR genes present in an individual may be classified into A and/or B haplotypes, which respectively have a more inhibitory role or a more activating role on the function of the NK cell. Family studies have been used to ascertain the make-up of these haplotypes, inclusion of allele typing enabling determination of whether one or two copies of a particular gene is present. In addition to genetic diversification the KIR gene complex shows differences at the functional level with different alleles having different protein expression levels and different avidity with their HLA ligand. PMID- 20028429 TI - Dissociated expression of natural killer 1.1 and T-cell receptor by invariant natural killer T cells after interleukin-12 receptor and T-cell receptor signalling. AB - Invariant (i) natural killer T (NKT) cells become undetectable after stimulation with alpha-galactosylceramide (alpha-GalCer) or interleukin (IL)-12. Although down-modulation of surface T-cell receptor (TCR)/NKR-P1C (NK1.1) expression has been shown convincingly after stimulation with alpha-GalCer, it is unclear whether this also holds true for IL-12 stimulation. To determine whether failure to detect iNKT cells after IL-12 stimulation is caused by dissociation/internalization of TCR and/or NKR-P1C, or by block of de novo synthesis of these molecules, and to examine the role of IL-12 in the disappearance of iNKT cells after stimulation with alpha-GalCer, surface (s)/cytoplasmic (c) protein expression, as well as messenger RNA (mRNA) expression of TCR/NKR-P1C by iNKT cells after stimulation with alpha-GalCer or IL 12, and the influence of IL-12 neutralization on the down-modulation of sTCR/sNKR P1C expression by iNKT cells after stimulation with alpha-GalCer were examined. The s/cTCR(+ )s/cNKR-P1C(+) iNKT cells became undetectable after in vivo administration of alpha-GalCer, which was partially prevented by IL-12 neutralization. Whereas s/cNKR-P1C(+) iNKT cells became undetectable after in vivo administration of IL-12, s/cTCR(+) iNKT cells were only marginally affected. mRNA expression of TCR/NKR-P1C remained unaffected by alpha-GalCer or IL-12 treatment, despite the down-modulation of cTCR and/or cNKR-P1C protein expression. By contrast, cTCR(+ )cNKR-P1C(+) sTCR(-) sNKR-P1C(-) iNKT cells and cNKR-P1C(+) sNKR-P1C(-) iNKT cells were detectable after in vitro stimulation with alpha-GalCer and IL-12, respectively. Our results indicate that TCR and NKR P1C expression by iNKT cells is differentially regulated by signalling through TCR and IL-12R. They also suggest that IL-12 participates, in part, in the disappearance of iNKT cells after stimulation with alpha-GalCer by down modulating not only sNKR-P1C, but also sTCR. PMID- 20028431 TI - Long-term exposure to indoor air pollution and wheezing symptoms in infants. AB - Long-term exposure to air pollution is suspected to cause recurrent wheeze in infants. The few previous studies have had ambiguous results. The objective of this study was to estimate the impact of measured long-term exposure to indoor air pollution on wheezing symptoms in infants. We monitored wheezing symptoms in diaries for a birth cohort of 411 infants. We measured long-term exposure to nitrogen oxides (NO(x)), NO(2), formaldehyde, PM(2.5) and black smoke in the infants' bedrooms and analyzed risk associations during the first 18 months of life by logistic regression with the dichotomous end-point 'any symptom-day' (yes/no) and by standard linear regression with the end-point 'number of symptom days'. The results showed no systematic association between risk for wheezing symptoms and the levels of these air pollutants with various indoor and outdoor sources. In conclusion, we found no evidence of an association between long-term exposure to indoor air pollution and wheezing symptoms in infants, suggesting that indoor air pollution is not causally related to the underlying disease. Practical Implications Nitrogen oxides, formaldehyde and fine particles were measured in the air in infants' bedrooms. The results showed no evidence of an association between long-term exposure and wheezing symptoms in the COPSAC birth cohort. PMID- 20028432 TI - Characteristics of indoor/outdoor particulate pollution in urban and rural residential environment of Pakistan. AB - Particulate pollution has emerged as a serious environmental health concern in Pakistan. The use of biomass fuels in traditional stoves produces high levels of indoor air pollutants. In Pakistan, 94% of rural and 58% of urban households depend on biomass fuel. This study investigates variations in indoor/outdoor concentrations of particulate matter during various activities for three different micro-environments in Pakistan. At a rural site, the average indoor/outdoor ratios for PM(10), PM(2.5), and PM(1), in kitchens using biomass fuels were 3.80, 4.36, and 4.11, respectively. A large variation was recorded in the mass concentration of particulate matter during cooking with concentrations in the range 4000-8555 microg/m(3). In a living room at a rural site, the average indoor/outdoor ratios for PM(10), PM(2.5), and PM(1) were 1.74, 2.49, and 3.01, respectively. At the urban site, the average indoor/outdoor ratios for the same size fractions were 1.71, 2.88, and 3.47, respectively. Cooking, cleaning and smoking were identified as principal contributors to the high indoor levels of particulate matter. This study showed considerably high concentrations of particulate matter, particularly in kitchens using biomass fuels, as compared to living areas. Thus women and children face the greatest exposure due to the amount of time they spend in the kitchen. PRACTICAL IMPLICATIONS: In the developing world, particulate air pollution, both indoor and outdoor, is a substantial health hazard to the public. The very high concentrations of particulate matter in both rural and urban sites, particularly in kitchens using biomass fuels, emphasize the severity of this issue in Pakistan. Women and children are extensively at risk due to amount of time spent in kitchens. This state of affairs calls for a large-scale intervention to reduce the exposure to indoor air pollution. PMID- 20028433 TI - Characterizing exhaled airflow from breathing and talking. AB - The exhaled air of infected humans is one of the prime sources of contagious viruses. The exhaled air comes from respiratory events such as the coughing, sneezing, breathing and talking. Accurate information on the thermo-fluid characteristics of the exhaled airflow can be important for prediction of infectious disease transmission. The present study developed a source model to provide the thermo-fluid conditions of the exhaled air from the breathing and talking processes. The source model is a set of equations obtained from the measurements of the flow rate, flow direction, and area of mouth/nose opening with human subjects. It was found that the exhaled flow rate over time can be represented as a sinusoidal function for breathing and a constant for talking. The flow rates can be calculated by physiological parameters of a subject. The direction of the exhalation jet did not vary much between subjects and the area of mouth/nose opening could be regarded as a constant. Though the mouth/nose opening size varied among subjects, they were not correlated with the physiological parameters of the subjects. If combined with appropriate virus and droplet distribution information, the model can be used to describe the disease source due to breathing and talking. PRACTICAL IMPLICATIONS: Accurate prediction of airborne disease transmission, and the infection prone zones, can aid in identifying and implementing the control strategies. With the recent advancements, Computational Fluid Dynamics (CFD) has become a powerful tool in predicting the disease transmission. Accurate prediction of the transmission by these CFD simulations requires information on sources and sinks of infectious viruses and models for dispersion of these viruses. The exhaled air of an infected human is one of the prime sources of disease viruses. In the present study, measurements of the flow were conducted on human subjects to develop models for the flow boundary conditions for the exhalation and inhalation during breathing and talking. PMID- 20028434 TI - Association between indoor exposure to semi-volatile organic compounds and building-related symptoms among the occupants of residential dwellings. AB - The aim of this study was to evaluate the levels of semi-volatile compounds (SVOCs) in residential detached houses in Sapporo, Japan, and whether exposure to these SVOCs was associated with the development of building-related symptoms named 'sick house syndrome' (SHS). The definition of SHS is fundamentally the same as that of the sick building syndrome (SBS). The presence of symptoms of SHS was evaluated using a validated self-administered questionnaire. Surveys and samplings of air and house dust in 41 dwellings were performed from October 2006 to January 2007, and 134 occupants responded to questionnaires. Samples were analyzed to quantify the concentrations of eight plasticizers, eleven phosphate triester flame retardants, two alkyl phenols used as anti-oxidants, and one organochlorine synergist called s-421, by gas chromatography-mass spectrometry and gas chromatography-flame photometry. The compounds frequently detected were di-n-butylphthalate, di(2-ethylhexyl)phthalate (DEHP), and dibutylhydroxytoluene in air, and DEHP and tris(2-butoxyethyl)phosphate (TBEP) in dust. Tributylphosphate was strongly and directly associated with mucosal symptoms of SHS; s-421 was also directly associated with mucosal symptoms of SHS. On the contrary, some chemicals such as diethylphthalate and TBEP were inversely associated with SHS. In future studies, we plan to assess these associations in a larger population. PRACTICAL IMPLICATIONS: This study suggests that it may be possible to reduce building-related symptoms by altering exposure to certain SVOCs, such as tributylphosphate commonly found in ceiling and wall coverings and s-421 used as a synergist for pyrethroids. The association between SHS and s-421 suggested that use of pyrethroid insecticides could elicit symptoms of SHS. However, further studies are necessary to test the associations observed in the present study and to examine whether the SVOCs associated with symptoms are causative agents or simply surrogates for some other factor that is causing the symptoms. PMID- 20028435 TI - Investigating ozone-induced decomposition of surface-bound permethrin for conditions in aircraft cabins. AB - The reaction of ozone with permethrin can potentially form phosgene. Published evidence on ozone levels and permethrin surface concentrations in aircraft cabins indicated that significant phosgene formation might occur in this setting. A derivatization technique was developed to detect phosgene with a lower limit of detection of 2 ppb. Chamber experiments were conducted with permethrin-coated materials (glass, carpet, seat fabric, and plastic) exposed to ozone under cabin relevant conditions (150 ppb O(3), 4.5/h air exchange rate, <1% relative humidity, 1700 ng/cm(2) of permethrin). Phosgene was not detected in these experiments. Reaction of ozone with permethrin appears to be hindered by the electron-withdrawing chlorine atoms adjacent to the double bond in permethrin. Experimental results indicate that the upper limit on the reaction probability of ozone with surface-bound permethrin is approximately 10(-7). Extrapolation by means of material-balance modeling indicates that the upper limit on the phosgene level in aircraft cabins resulting from this chemistry is approximately 1 microg/m(3) or approximately 0.3 ppb. It was thus determined that phosgene formation, if it occurs in aircraft cabins, is not likely to exceed relevant, health-based phosgene exposure guidelines. PRACTICAL IMPLICATIONS: Phosgene formation from ozone-initiated oxidation of permethrin in the aircraft cabin environment, if it occurs, is estimated to generate levels below the California Office of Environmental Health Hazard Assessment acute reference exposure level of 4 microg/m(3) or approximately 1 ppb. PMID- 20028436 TI - Characterization of recombinant prolyl aminopeptidase from Aspergillus oryzae. AB - AIMS: Prolyl aminopeptidase (PAP) degrades only amino-terminal proline from peptides. The food-grade fungus Aspergillus oryzae produces this enzyme only in small amounts. In this paper, we present efficient production of recombinant PAP with an overexpression system of A. oryzae and characterization of its biochemical properties. METHODS AND RESULTS: The gene encoding PAP was overexpressed as a His-tag fusion protein under a taka-amylase gene (amyB) promoter with a limited expressing condition in A. oryzae. The PAP activity in the mycelia grown in rich medium containing glucose (repressing condition) was twice that in starch (inducing condition). The enzyme prepared as cell-free extract was partially purified through two-step column chromatography. The PAP was estimated to be a hexameric protein and exhibited salt tolerance against NaCl of up to 4 mol l(-1). CONCLUSIONS: Aspergillus oryzae PAP was produced under the repressing condition of amyB promoter in a PAP-overexpressing strain and purified 1800-folds. Overproduction of PAP under promoter-inducing conditions led to an increase in inactive PAP, possibly because of irregular folding. SIGNIFICANCE AND IMPACT OF THE STUDY: PAP with a high specific activity and salt tolerance may be used effectively in the manufacturing processes of fermented foods. PMID- 20028437 TI - Design and optimization of a novel reverse transcription linear-after-the exponential PCR for the detection of foot-and-mouth disease virus. AB - AIMS: A novel molecular assay for the detection of foot-and-mouth disease virus (FMDV) was developed using linear-after-the-exponential polymerase chain reaction (LATE-PCR). METHODS AND RESULTS: Pilot experiments using synthetic DNA targets demonstrated the ability of LATE-PCR to quantify initial target concentration through endpoint detection. A two-step protocol involving reverse transcription (RT) followed by LATE-PCR was then used to confirm the ability of the assay to detect FMDV RNA. Finally, RT and LATE-PCR were combined in a one-step duplex assay for co-amplification of an FMDV RNA segment and an internal control comprised of an Armored RNA. In that form, each of the excess primers in the reaction mixture hybridize to their respective RNA targets during a short pre incubation, then generate cDNA strands during a 3-min RT step at 60 degrees C, and the resulting cDNA is amplified by LATE-PCR without intervening sample processing. CONCLUSIONS: The RT-LATE-PCR assay generates fluorescent signals at endpoint that are proportional to the starting number of RNA targets and can detect a range of sequence variants using a single mismatch-tolerant probe. SIGNIFICANCE AND IMPACT OF THE STUDY: In addition to offering improvements over current laboratory-based molecular diagnostic assays for FMDV, this new assay is compatible with a novel portable ('point-of-care') device, the BioSeeq II, designed for the rapid diagnosis of FMD in the field. PMID- 20028438 TI - High hydrostatic pressure resistance of Campylobacter jejuni after different sublethal stresses. AB - AIMS: To study the development of resistance responses in Campylobacter jejuni to high hydrostatic pressure (HHP) treatments after the exposure to different stressful conditions that may be encountered in food-processing environments, such as acid pH, elevated temperatures and cold storage. METHODS AND RESULTS: Campylobacter jejuni cells in exponential and stationary growth phase were exposed to different sublethal stresses (acid, heat and cold shocks) prior to evaluate the development of resistance responses to HHP. For exponential-phase cells, neither of the conditions tested increased nor decreased HHP resistance of C. jejuni. For stationary-phase cells, acid and heat adaptation-sensitized C. jejuni cells to the subsequent pressure treatment. On the contrary, cold-adapted stationary-phase cells developed resistance to HHP. CONCLUSIONS: Whereas C. jejuni can be classified as a stress sensitive micro-organism, our findings have demonstrated that it can develop resistance responses under different stressing conditions. The resistance of stationary phase C. jejuni to HHP was increased after cells were exposed to cold temperatures. SIGNIFICANCE AND IMPACT OF THE STUDY: The results of this study contribute to a better knowledge of the physiology of C. jejuni and its survival to food preservation agents. Results here presented may help in the design of combined processes for food preservation based on HHP technology. PMID- 20028439 TI - Production of 4-ethylphenol from 4-hydroxycinnamic acid by Lactobacillus sp. isolated from a swine waste lagoon. AB - AIMS: To determine the source material and the responsible micro-organisms of 4 ethylphenol production in swine lagoon sediment slurries. METHODS AND RESULTS: Swine lagoon sediment was blended and incubated with tryptone-yeast extract broth containing 10 mmol l(-1) each of 4-hydroxycinnamic as well as other phenolics. 4 Ethylphenol was only produced from 4-hydroxycinnamic acid. Denaturing gradient gel electrophoresis analysis indicated that the microbial community was substantially altered from inclusion of 4-hydroxycinnamic acid. Serial dilutions and selective plating were employed to isolate a culture capable of 4 hydroxycinnamic acid conversion to 4-ethylphenol. Morphological and ribosomal gene analysis indicated the isolate to be a Lactobacillus sp., Lactobacillus sp. pep8 converted 4-vinylphenol, but not 4-hydroxy-3-phenylpropionate, to 4 ethylphenol and did not convert 4-hydroxy-3-methoxycinnamic acid to 4-ethyl-2 methoxyphenol. A p-coumaric acid decarboxylase (pdc) gene identical to Lact. plantarum pdc genes was cloned from Lactobacillus sp. pep8. CONCLUSIONS: Swine lagoon sediments produce 4-ethylphenol from 4-hydroxycinnamic acid due, in part, to the activity of Lactobacillus spp. SIGNIFICANCE AND IMPACT OF THE STUDY: 4 Ethylphenol, a malodourant of swine and beef wastes, is generated from plant materials by indigenous lactobacilli suggesting that altering the amount dietary of plant material may influence levels of 4-ethylphenol in the wastes. PMID- 20028440 TI - Application of quantitative reverse-transcription PCR (qRT-PCR) for the determination of the total viable count (TVC) on meat samples. AB - AIM: To apply a quantitative reverse-transcription PCR (qRT-PCR) method to determine the total viable count (TVC) on meat samples. METHODS AND RESULTS: Using two sets of primers to target the ribonuclease-P (RNase P) RNA transcripts of Gram-positive and Gram-negative bacteria, standard curves were generated using the LightCycler 2.0 instrument (Roche Diagnostics). RNA standards were extracted from known cell numbers and subsequently converted to cDNA for the construction of standard curves for quantification of the TVC of beef carcass swabs (n = 60) and beef (n = 30), chicken (n = 50) and pork (n = 49) pieces. A high correlation between the standard plate count method and the qRT-PCR was observed for beef swabs (R(2) = 0.93) and beef pieces (R(2) = 0.82). The correlation coefficient for chicken pieces and pork pieces were R(2) = 0.34 and 0.55, respectively. Using beef pieces (n = 13), an interlaboratory study was conducted and each participating laboratory (n = 3) found a reasonable degree of agreement between the cultural method and the PCR method. CONCLUSIONS: The qRT-PCR assay used in this study can enumerate the total bacteria on beef samples with a high degree of accuracy. SIGNIFICANCE AND IMPACT OF THE STUDY: The qRT-PCR method may have the potential to be applied to various sample types as an alternative rapid method for determining TVCs; however, further validation would be required. PMID- 20028441 TI - A novel mutation in the FERMT1 gene in a Spanish family with Kindler's syndrome. PMID- 20028442 TI - Tattoo removal by non-professionals--medical and forensic considerations. AB - Tattoos are common phenomena in the western world and the demand for their removal has become widespread in the recent years. The introduction of quality switched lasers has provided an effective removal method that is increasingly being exploited by tattoo studios themselves. Dermatologists, however, are frequently confronted with complications and side effects of tattoo removals that were performed by non-professionals. The objective of this study is to assess potential risks and pitfalls of laser tattoo removal when performed by medical laymen. The methods followed are systematic and evidence-based review of the literature. Four major problem areas were identified: rare but potentially severe allergic or toxic effects of decomposition products of the tattoo pigments; bodily harm caused by out-of-specification usage of the laser devices; malignant disease that is obscured within the area of a tattoo and requires meticulous dermatological diagnosis; and insufficient pre-operative consultation of patients about the risks, side effects and realistic expectations on the therapeutic outcome. We came to a conclusion that tattoo laser removal by medical laymen is unacceptable from the point of view of patient safety and the laws need to ban this practice swiftly. PMID- 20028443 TI - Non-ablative 1550-nm erbium-glass and ablative 10 600-nm carbon dioxide fractional lasers for acne scars: a randomized split-face study with blinded response evaluation. AB - BACKGROUND: Non-ablative 1550-nm erbium-doped fractional photothermolysis systems (FPS) and 10 600-nm carbon dioxide fractional laser systems (CO(2) FS) have been effectively used to treat scars. OBJECTIVE: We compared the efficacy and safety of single-session treatments of FPS and CO(2) FS for acne scars through a randomized, split-face, evaluator-blinded study. METHODS: Eight patients with acne scars were enrolled in this study. Half of each subject's face was treated with FPS and the other half was treated with CO(2) FS. We used a quartile grading scale for evaluations. RESULTS: At 3 months after the treatment, the mean grade of improvement based on clinical assessment was 2.0 +/- 0.5 for FPS and 2.5 +/- 0.8 for CO(2) FS. On each side treated by FPS and CO(2) FS, the mean duration of post-therapy crusting and scaling was 2.3 and 7.4 days respectively and that of post-therapy erythema was 7.5 and 11.5 days respectively. The mean VAS pain score was 3.9 +/- 2.0 with the FPS and 7.0 +/- 2.0 with the CO(2) FS. CONCLUSION: We demonstrated the efficacy and safety of single-session acne scar treatment using FPS and CO(2) FS in East Asian patients. We believe that our study could be used as an essential reference when choosing laser modalities for scar treatment. PMID- 20028444 TI - Latanoprost in the treatment of eyelash alopecia in alopecia areata universalis. AB - OBJECTIVES: The aim of this study was to test the efficacy of latanoprost in eyelash alopecia areata (AA). DESIGN: This study is a 2-year prospective, non blinded, non-randomized, bilateral eyelash alopecia controlled study. SETTING: The setting of this study was Trichology Unit, Virgen Macarena University Hospital, Seville, Spain. PATIENTS: We conducted a survey of 54 subjects with AA universalis treated with the protocol of the Trichology Unit of our Department. Control group comprised 10 subjects who received injections of 0.5 mg/cm(2) of triamcinolone acetonide (TAC) in their eyebrows and 1 mg/cm(2) of TAC injections in affected scalp. The treatment group included 44 subjects who received the same treatment as the control group in scalp and eyebrows but they also applied a drop of latanoprost 0.005% (50 microg/mL) ophthalmic solution in their eyelid margins every night. Subjects were reviewed every 3 months for 2 years. RESULTS: Forty subjects finished the study and four subjects were lost to follow-up. In the treatment arm of this study, the course was well tolerated and uncomplicated. Both investigators and patients evaluated the regrowth. The results we obtained were: complete regrowth in 17.5%, moderate regrowth in 27.5%, slight regrowth in 30% and without response in 25%. Moderate and total regrowth constituted a cosmetically acceptable response. The therapy was continuous and the response remained without any side effects. No patients had cosmetically acceptable eyelash regrowth in the control group. CONCLUSIONS: Latanoprost may be an effective drug in the treatment of eyelash AA because it induces acceptable responses (total and moderate) in 45% of the patients. A formal, blinded prospective unilateral controlled study will permit further understanding about this promising therapeutic agent for eyelash AA. PMID- 20028445 TI - Oral verrucous carcinoma treated with carbon dioxide laser. PMID- 20028446 TI - Dermoscopic rainbow pattern in basal cell carcinoma. PMID- 20028447 TI - Efficacy of amorolfine nail lacquer for the prophylaxis of onychomycosis over 3 years. AB - BACKGROUND: Standard treatment for onychomycosis often results in less than half of subjects achieving disease-free nails. Onychomycosis is even more challenging to treat as relapses and re-infections are common. OBJECTIVE: To determine if a prophylactic effect exists when a treatment with amorolfine nail lacquer (ANL), with half the frequency of the standard regimen, is instituted following successful treatment of dermatophytic toenail onychomycosis with matrix involvement. METHODS: Efficacy and safety of a group treated with ANL (once every 2 weeks) were compared with that of an untreated group in a 36-month (3 years), single-centre, randomized, open-label, comparison study. Subjects to be included in the study were required to be cured of confirmed onychomycosis with matrix involvement after an initial treatment with either ANL + oral terbinafine or oral terbinafine alone in a previous study. Prophylaxis of onychomycosis was assessed by global recurrence rate, confirmed onychomycosis, clinical recurrence and mycological recurrence. RESULTS: A total of 52 subjects were enrolled (26 in each group) in the study. Throughout the study, recurrences occurred more quickly in the untreated group compared with that in the ANL group. Statistically significant differences were observed at month 12 (ANL, 8.3%; untreated, 31.8%; P = 0.047). At endpoint, 70.8% of the subjects treated with ANL remained cured compared to 50% in the untreated group (P = 0.153). Recurrence was delayed by nearly 200 days for the ANL group compared with that of the untreated group. Amorolfine was safe and well tolerated during the study, with no treatment related adverse events. CONCLUSION: These results suggest that amorolfine nail lacquer may be effective and is safe for use as a prophylactic treatment for the recurrence of onychomycosis. PMID- 20028448 TI - Photodynamic therapy with 5-aminolevulinic acid in actinic cheilitis: an 18-month clinical and histological follow-up. AB - BACKGROUND: Actinic cheilitis (AC) may bear the initial and superficial changes of actinically induced squamous cell carcinoma (SCC) and may progress into fully developed SCCs. Early and effective treatment is important. OBJECTIVE: To assess the clinical and histological long-term outcome in AC after two ALA-PDT sessions. METHODS: Patients with histologically proven grade 1 and 2 AC received two ALA PDT sessions at 2 weeks interval. Subjects with complete clinical response at 3 months were evaluated further clinically and histologically at months 6, 12 and 18. Long-term study outcome was defined as clinical and histological AC recurrence among patients with complete clinical response 3 months after treatment. Cosmetic outcome was assessed by the investigators at the final follow up visit at 18 months. RESULTS: Of the 40 patients enrolled, 38 completed the study. Complete clinical response at 3 months was achieved in 26 patients. At 6 months, clinical and histological recurrence occurred in three patients and at 12 months, one more patient showed clinical and histological recurrence. At 18 months, overall clinical recurrence rate was 15.38% (4/26), while overall histological recurrence rate was 34.61% (9/26). Cosmetic outcome was rated as excellent in more than 80% of evaluated cases. CONCLUSION: PDT represents a moderately effective treatment modality in AC. Optimization of treatment procedure and protocols is still needed for higher response rates to be achieved. Moreover, the high treatment cost should be given consideration. Further long term follow-up studies are needed for assessment of clinical and histological very late recurrences that could be expected after PDT. PMID- 20028449 TI - Acquired erythroderma in adults: a clinical and prognostic study. AB - BACKGROUND: Erythroderma is a severe syndrome and prognostic studies are rare in the literature. OBJECTIVES: Through a retrospective study of erythroderma in adults, we have analysed epidemiological and clinical data and precised the relevant aetiologies and survival in our patients. METHODS: This study was performed at the Department of Dermatology of Charles Nicolle Hospital of Tunis (1995-2007) including 82 cases of acquired erythroderma (>16 years). We have recorded epidemio-clinical, biological and histological data, treatment and outcome. Clinical-histological correlation was analysed [kappa coefficient (kappa)]. Follow-up time and disease-free survival time were calculated as were Kaplan-Meier estimates of overall survival and relapse-free survival for some aetiologies. RESULTS: Erythroderma represented 0.44 per thousand of all dermatoses with an age of 55.13 +/- 18.16 and no sex predilection. Psoriasis was the predominant aetiology (32.9%) with a median duration of 6.75 years and previous one or more episodes of erythroderma. Psoriasis was significantly associated with pruritus (P = 0.0001), pachyonychia (P = 0.00001), palmoplantar keratoderma (P = 0.0001) and hypereosinophilia (P = 0.008). The latter is then not specific for drug induced erythroderma (P = 0.004). Carbamazepine (27.8%) and penicillin (22.2%) were the most implicated drugs. Positive Clinical-histological correlation was found in 77% of cases (kappa = 0.753). Relapse was seen in all aetiologies, but drug reactions and had occurred in the first 3 years in 90% of them. Mortality rate was 11.3 per 1000 patients-years. CONCLUSIONS: Our study illustrates the severity of erythroderma. It alters heavily the quality of life of patients which is initially altered by the pre-existent dermatosis. It may be life threatening as mortality rate is high. PMID- 20028450 TI - Involvement of the AT1 receptor subtype in the effects of angiotensin IV and LVV haemorphin 7 on hippocampal neurotransmitter levels and spatial working memory. AB - Intracerebroventricular (i.c.v.) administration of angiotensin IV (Ang IV) or Leu Val-Val-haemorphin 7 (LVV-H7) improves memory performance in normal rats and reverses memory deficits in rat models for cognitive impairment. These memory effects were believed to be mediated via the putative 'AT4 receptor'. However, this binding site was identified as insulin-regulated aminopeptidase (IRAP). Correspondingly, Ang IV and LVV-H7 were characterised as IRAP inhibitors. This study investigates whether and how IRAP may be involved in the central effects of Ang IV and LVV-H7. We determined the effects of i.c.v. administration of Ang IV or LVV-H7 on hippocampal neurotransmitter levels using microdialysis in rats. We observed that Ang IV modulates hippocampal acetylcholine levels, whereas LVV-H7 does not. This discrepancy was reflected in the observation that Ang IV binds with micromolar affinity to the AT1 receptor whereas no binding affinity was observed for LVV-H7. Correspondingly, we demonstrated that the AT1 receptor is involved in the effects of Ang IV on hippocampal neurotransmitter levels and on spatial working memory in a plus maze spontaneous alternation task. However, the AT1 receptor was not involved in the spatial memory facilitating effect of LVV H7. Finally, we demonstrated that Ang IV did not diffuse to the hippocampus following i.c.v. injection, suggesting an extrahippocampal site of action. We propose that AT1 receptors are implicated in the neurochemical and cognitive effects of Ang IV, whereas LVV-H7 may mediate its effects via IRAP. PMID- 20028451 TI - Pathogenic cysteine mutations affect progranulin function and production of mature granulins. AB - Frontotemporal dementia with ubiquitin-positive inclusions (FTLD-U) can be caused by mutations in the progranulin gene (GRN). Progranulin (PGRN) is a cysteine-rich growth factor, which is proteolytically cleaved by elastase to produce several granulins (GRNs). All FTLD-U mutations in GRN characterized to date result in reduced secreted PGRN protein. We recently reported a Spanish family with progressive non-fluent aphasia and dementia in which a novel C521Y mutation segregates with disease. A second cysteine mutation (C139R) has also been reported to be disease specific. Allele-specific mRNA expression assays in brain reveal that the C521Y mutant allele is expressed at similar levels to the wild type allele. Furthermore, plasma PGRN levels in C521Y carriers are comparable with non-carrier family relatives, suggesting that the mutation does not affect PGRN protein expression and secretion in vivo. Despite normal PGRN levels C521Y and C139R mutant GRNs show reduced neurite growth-stimulating activity in vitro. Further study revealed that these mutations also cause impaired cleavage of PGRN by elastase. Our data suggest that these mutations affect the function of full length PGRN as well as elastase cleavage of PGRN into GRNs, leading to neurodegeneration. PMID- 20028452 TI - Central and peripheral consequences of the chronic blockade of CB1 cannabinoid receptor with rimonabant or taranabant. AB - The endocannabinoid system plays a crucial role in the pathophysiology of obesity. However, the clinical use of cannabinoid antagonists has been recently stopped because of its central side-effects. The aim of this study was to compare the effects of a chronic treatment with the CB(1) cannabinoid antagonist rimonabant or the CB(1) inverse agonist taranabant in diet-induced obese female rats to clarify the biological consequences of CB(1) blockade at central and peripheral levels. As expected, chronic treatment with rimonabant and taranabant reduced body weight and fat content. Interestingly, a decrease in the number of CB(1) receptors and its functional activity was observed in all the brain areas investigated after chronic taranabant treatment in both lean and obese rats. In contrast, chronic treatment with rimonabant did not modify the density of CB(1) cannabinoid receptor binding, and decreased its functional activity to a lower degree than taranabant. Six weeks after rimonabant and taranabant withdrawal, CB(1) receptor density and activity recovered to basal levels. These results reveal differential adaptive changes in CB(1) cannabinoid receptors after chronic treatment with rimonabant and taranabant that could be related to the central side-effects reported with the use of these cannabinoid antagonists. PMID- 20028453 TI - The more you have, the less you get: the functional role of inflammation on neuronal differentiation of endogenous and transplanted neural stem cells in the adult brain. AB - The differentiation of neural stem cells toward a neuronal phenotype is determined by the extracellular and intracellular factors that form the neurogenic niche. In this review, we discuss the available data on the functional role of inflammation and in particular, pro- and anti-inflammatory cytokines, on neuronal differentiation from endogenous and transplanted neural stem/progenitor cells. In addition, we discuss the role of microglial cell activation on these processes and the fact that microglial cell activation is not univocally associated with a pro-inflammatory milieu. We conclude that brain cytokines could be regarded as part of the endogenous neurogenic niche. In addition, we propose that accumulating evidence suggests that pro-inflammatory cytokines have a negative effect on neuronal differentiation, while anti-inflammatory cytokines exert an opposite effect. The clarification of the functional role of cytokines on neuronal differentiation will be relevant not only to better understand adult neurogenesis, but also to envisage complementary treatments to modulate cytokine action that could increase the therapeutic benefit of future progenitor/stem cell based therapies. PMID- 20028455 TI - Mesenchymal stem cell-secreted superoxide dismutase promotes cerebellar neuronal survival. AB - It has been postulated that bone marrow-derived mesenchymal stem cells (MSCs) might be effective treatments for neurodegenerative disorders either by replacement of lost cells by differentiation into functional neural tissue; modulation of the immune system to prevent further neurodegeneration; and/or provision of trophic support for the diseased nervous system. Here we have performed a series of experiments showing that human bone marrow-derived MSCs are able to protect cultured rodent cerebellar neurons, and specifically cells expressing Purkinje cell markers, against either nitric oxide exposure or withdrawal of trophic support via cell-cell contact and/or secretion of soluble factors, or through secretion of soluble factors alone. We have demonstrated that MSCs protect cerebellar neurons against toxic insults via modulation of both the phosphatidylinositol 3-kinase/Akt and MAPK pathways and defined superoxide dismutase 3 as a secreted active antioxidant biomolecule by which MSCs modulate, at least in part, their neuroprotective effect on cerebellar cells in vitro. Together, the results demonstrate new and specific mechanisms by which MSCs promote cerebellar neuronal survival and add further evidence to the concept that MSCs may be potential therapeutic agents for neurological disorders involving the cerebellum. PMID- 20028456 TI - Plumbagin, a novel Nrf2/ARE activator, protects against cerebral ischemia. AB - Many phytochemicals function as noxious agents that protect plants against insects and other damaging organisms. However, at subtoxic doses, the same phytochemicals may activate adaptive cellular stress response pathways that can protect cells against a variety of adverse conditions. We screened a panel of botanical pesticides using cultured human and rodent neuronal cell models, and identified plumbagin as a novel potent activator of the nuclear factor E2-related factor 2 (Nrf2)/antioxidant response element (ARE) pathway. In vitro, plumbagin increases nuclear localization and transcriptional activity of Nrf2, and induces the expression of the Nrf2/ARE-dependent genes, such as heme oxygenase 1 in human neuroblastoma cells. Plumbagin specifically activates the Nrf2/ARE pathway in primary mixed cultures from ARE-human placental alkaline phosphatase reporter mice. Exposure of neuroblastoma cells and primary cortical neurons to plumbagin provides protection against subsequent oxidative and metabolic insults. The neuroprotective effects of plumbagin are abolished by RNA interference-mediated knockdown of Nrf2 expression. In vivo, administration of plumbagin significantly reduces the amount of brain damage and ameliorates-associated neurological deficits in a mouse model of focal ischemic stroke. Our findings establish precedence for the identification and characterization of neuroprotective phytochemicals based upon their ability to activate adaptive cellular stress response pathways. PMID- 20028457 TI - Nicotine self-administration diminishes stress-induced norepinephrine secretion but augments adrenergic-responsiveness in the hypothalamic paraventricular nucleus and enhances adrenocorticotropic hormone and corticosterone release. AB - Chronic nicotine self-administration augments the thalamo-pituitary-adrenal (HPA) responses to stress. Altered neuropeptide expression within corticotropin releasing factor (CRF) neurons in the hypothalamic paraventricular nucleus (PVN) contributes to this enhanced HPA response to stress. Herein, we determined the role of norepinephrine, a primary regulator of CRF neurons, in the responses to footshock during nicotine self-administration. On day 12-15 of self administration, microdialysis showed nicotine reduced PVN norepinephrine release by footshock (< 50% of saline). Yet, the reduction in footshock-induced adrenocorticotropic hormone and corticosterone secretion because of intra-PVN prazosin (alpha(1) adrenergic antagonist) was significantly greater in rats self administering nicotine (2-fold) than saline. Additionally, PVN phenylephrine (alpha(1) agonist) stimulated adrenocorticotropic hormone and corticosterone release to a similar extent in unstressed rats self-administering nicotine or saline. Nicotine self-administration also decreased footshock-induced c-Fos expression in the nucleus of the solitary tract-A2/C2 catecholaminergic neurons that project to the PVN. Therefore, footshock-induced nucleus of the solitary tract activation and PVN norepinephrine input are both attenuated by nicotine self-administration, yet PVN CRF neurons are more responsive to alpha(1) stimulation, but only during stress. This plasticity in noradrenergic regulation of PVN CRF neurons provides a new mechanism contributing to the HPA sensitization to stress by nicotine self-administration and smoking. PMID- 20028459 TI - Minireview: Invasive fungal infection complicating acute Plasmodium falciparum malaria. AB - Malaria is the most important parasitic infection in people, affecting 5-10% of the world's population with more than two million deaths a year. Whereas invasive bacterial infections are not uncommon during severe Plasmodium falciparum malaria, only a few cases of opportunistic fungal infections have been reported. Here, we present a fatal case of disseminated hyalohyphomycosis associated with acute P. falciparum malaria in a non-immune traveller, review the cases reported in the literature and discuss the theoretical foundations for the increased susceptibility of non-immune individuals with severe P. falciparum malaria to opportunistic fungal infections. Apart from the availability of free iron as sequelae of massive haemolysis, tissue damage, acidosis and measures of advanced life support, patients with complicated P. falciparum malaria also are profoundly immunosuppressed by the organism's interaction with innate and adaptive host immune mechanisms. PMID- 20028458 TI - Nanomolar vitamin E alpha-tocotrienol inhibits glutamate-induced activation of phospholipase A2 and causes neuroprotection. AB - Our previous works have elucidated that the 12-lipoxygenase pathway is directly implicated in glutamate-induced neural cell death, and that such that toxicity is prevented by nM concentrations of the natural vitamin E alpha-tocotrienol (TCT). In the current study we tested the hypothesis that phospholipase A(2) (PLA(2)) activity is sensitive to glutamate and mobilizes arachidonic acid (AA), a substrate for 12-lipoxygenase. Furthermore, we examined whether TCT regulates glutamate-inducible PLA(2) activity in neural cells. Glutamate challenge induced the release of [(3)H]AA from HT4 neural cells. Such response was attenuated by calcium chelators (EGTA and BAPTA), cytosolic PLA(2) (cPLA(2))-specific inhibitor (AACOCF(3)) as well as TCT at 250 nM. Glutamate also caused the elevation of free polyunsaturated fatty acid (AA and docosahexaenoic acid) levels and disappearance of phospholipid-esterified AA in neural cells. Furthermore, glutamate induced a time-dependent translocation and enhanced serine phosphorylation of cPLA(2) in the cells. These effects of glutamate on fatty acid levels and on cPLA(2) were significantly attenuated by nM TCT. The observations that AACOCF(3), transient knock-down of cPLA(2) as well as TCT significantly protected against the glutamate-induced death of neural cells implicate cPLA(2) as a TCT-sensitive mediator of glutamate induced neural cell death. This work presents first evidence recognizing glutamate-induced changes in cPLA(2) as a novel mechanism responsible for neuroprotection observed in response to nanomolar concentrations of TCT. PMID- 20028460 TI - Minireview: Malassezia infections in immunocompromised patients. AB - Malassezia spp. form part of the normal human cutaneous flora and are implicated in several mild, but recurrent cutaneous diseases, such as pityriasis versicolor, Malassezia folliculitis, seborrhoeic dermatitis, and, with lesser frequency, a range of other dermatological disorders. Malassezia spp. have also been associated with cutaneous and systemic diseases in immunocompromised patients including folliculitis, seborrhoeic dermatitis, catheter-related fungaemia and a variety of deeply invasive infections. In this review, we provide an overview of the epidemiology, risk factors, pathogenesis, clinical manifestations, diagnosis, treatment and outcome of cutaneous and invasive Malassezia infections in immunocompromised patients. PMID- 20028461 TI - Phospholipase and proteinase activities of Candida isolates from denture wearers. AB - The aim of the present study was to characterise phospholipase and proteinase activities of oral Candida isolates from 100 denture wearers and to study the relationship of these activities with denture stomatitis. Of 100 patients studied, 44 suffered from denture stomatitis. Specimens were collected by swabbing the denture and underlying mucosa. Isolates were previously identified by conventional mycological and genotypic methods. The phospholipase and proteinase activities were evaluated by agar plate methods. A total of 152 isolates were recovered from denture and underlying mucosa, including 101 Candida albicans, 18 Candida tropicalis, 14 Candida glabrata, 11 Candida guilliermondii, four Candida parapsilosis, two Saccharomyces cerevisiae and one isolate each of Candida dubliniensis and Candida krusei. Most C. albicans (97%) showed phospholipase activity; furthermore, the unique C. dubliniensis isolate showed a moderate phospholipase activity. The isolation of C. albicans (chi-square test, P = 0.0016) and phospholipase production by Candida spp. (chi-square test, P = 0.0213) was found to be significantly associated with denture stomatitis. Proteinase production was observed in <30% of isolates, and it was not related to the presence of denture stomatitis (P = 0.7675). Candida albicans isolates may produce both virulence factors, although the proteinase production was only observed in <30% of the isolates. Phospholipase production was exclusive of C. albicans and C. dubliniensis. PMID- 20028462 TI - Cladophialophora carrionii: an aetiological agent of cutaneous chromoblastomycosis from a non-endemic area, North India. AB - We present a case of infection due to Cladophialophora carrionii, an agent of Chromoblastomycosis in a 37-year-old Indian male. The patient developed a nodule as the lateral malleolus of his left leg. The lesion was successfully treated with surgical excision. Histopathologically, pigmented organisms were readily identified in tissue sections, and the cultural characteristics were these of Cladophialophora carrionii. PMID- 20028463 TI - Species distribution and antifungal susceptibility of blood Candida isolates at a tertiary hospital in southern Taiwan, 1999-2006. AB - The aim of this study was to evaluate the incidence of candidaemia, consumption of fluconazole and susceptibility of blood Candida isolates at a tertiary hospital. From January 1999 to September 2006, all candidaemic episodes were identified and available strains were evaluated for the susceptibilities of antifungal agents. Annual defined daily doses of antifungal agents were collected. There had been 909 Candida isolates detected from the bloodstream of 843 patients during the study period. Among them, 740 isolates were available for the susceptibilities of antifungal agents. The incidence density of candidaemia was 28 episodes per 10,000 patient-days. Species distribution of 909 isolates did not vary annually, but varied greatly in the units of the hospital. Candida parapsilosis was the more prominent (30.1%) isolate in the paediatric units, where C. tropicalis and C. glabrata were less common (12.3% and 1.4% respectively). Resistance rates for itraconazole, fluconazole and voriconazole were 6.9%, 3.8% and 3.8% respectively. There were 25 (3.4%) isolates resistant to amphotericin-B. Although fluconazole usage increased over time (r(2) = 0.45; P = 0.07), fluconazole resistance did not increase accordingly (P = 0.33). In our institution in which the incidence of candidaemia was high, fluconazole resistance among blood Candida isolates remained rare. PMID- 20028465 TI - Sudden vision loss caused by rhino-orbital zygomycosis in diabetic patients: case series. AB - Rhino-orbital zygomycosis is a life-threatening fungal infection generally occurring in patients with an underlying disorder, such as diabetes mellitus with ketoacidosis or with immunocompromising factors, although it may rarely appear in healthy individuals. The study has been undertaken to discuss the clinical presentation, pathogenesis, diagnostic work up and management of this rapidly progressive disease. Four male patients having uncontrolled diabetes and presenting with signs and symptoms of rhino-orbital zygomycosis were studied to illustrate the serious nature of the disease. All the four patients had rapidly deteriorating vision loss either unilateral or bilateral along with other nasal and orbital signs and symptoms. All the patients were put on liposomal amphotericin B and underwent orbital exenteration and pansinusectomy. One patient died, while the other three were successfully treated. Early diagnosis is critical in the prevention of morbidity and mortality associated with the disease. There is need for a high index of clinical suspicion in immunocompromised patients. Timely medical-surgical treatment proves extremely important for prognosis. PMID- 20028464 TI - Serum correlates of the placebo effect in irritable bowel syndrome. AB - BACKGROUND In diseases defined primarily by the subjective nature of patient self report, placebo effects can overwhelm the capacity of randomized controlled trials to detect medication-placebo differences. Moreover, it is unclear whether such placebo effects represent genuine psychobiological phenomena or just shifts in selective attention. Knowledge of predictors of the placebo response could improve the design of clinical trials and the delivery of personalized medical care. METHODS In patients with irritable bowel syndrome (IBS), a subset of our previous study that were randomized to placebo treatment (sham acupuncture) or no treatment group (waitlist), we tested an enriched panel of 10 serum biomarkers at the enrolment and the 3rd week of intervention, using a multiplex electrochemiluminescent immunoassay. KEY RESULTS More pronounced changes overtime in serum levels of osteoprotegerin (OPG) have been found in patients who received placebo treatment compared with the waitlist group (P = 0.039). Moreover, serum levels of OPG at baseline were found to be higher (P = 0.0167) in patients who subsequently achieved adequate relief (AR) of their IBS symptoms, independently of their treatment group. Besides, serum levels of TNF-related weak inducer of apoptosis (TWEAK) at baseline were also higher (P = 0.0144) in patients who reported AR and in particular in those who received the placebo treatment. CONCLUSIONS & INFERENCES These two measurable biological parameters associated with placebo, namely serum OPG and TWEAK, provide a proof of principle for discovering putative molecular signatures of placebo response in IBS and perhaps in other illnesses with patient self-reported outcomes. PMID- 20028466 TI - A rare cause of acute respiratory failure--allergic bronchopulmonary aspergillosis. AB - Allergic bronchopulmonary aspergillosis (ABPA) is a complex immune hypersensitivity reaction to Aspergillus fumigatus, usually complicating the course of patients with asthma and cystic fibrosis. The common radiological manifestations encountered are fleeting pulmonary opacities, bronchiectasis and mucoid impaction. Uncommon radiological findings encountered in ABPA include pulmonary masses, perihilar opacities simulating hilar adenopathy, miliary nodules and pleural effusions. Herein, we describe a 22-year-old female patient who presented with acute hypoxaemic respiratory failure secondary to left lung collapse, which necessitated rigid bronchoscopy for management. On further evaluation, she was diagnosed to have ABPA. This is the first documented report of ABPA presenting as acute hypoxaemic respiratory failure secondary to lung collapse. PMID- 20028467 TI - A dynamic model of nutrient uptake by root hairs. AB - Root hairs are known to be important in the uptake of sparingly soluble nutrients by plants, but quantitative understanding of their role in this is weak. This limits, for example, the breeding of more nutrient-efficient crop genotypes. We developed a mathematical model of nutrient transport and uptake in the root hair zone of single roots growing in soil or solution culture. Accounting for root hair geometry explicitly, we derived effective equations for the cumulative effect of root hair surfaces on uptake using the method of homogenization. Analysis of the model shows that, depending on the morphological and physiological properties of the root hairs, one of three different effective models applies. They describe situations where: (1) a concentration gradient dynamically develops within the root hair zone; (2) the effect of root hair uptake is negligibly small; or (3) phosphate in the root hair zone is taken up instantaneously. Furthermore, we show that the influence of root hairs on rates of phosphate uptake is one order of magnitude greater in soil than solution culture. The model provides a basis for quantifying the importance of root hair morphological and physiological properties in overall uptake, in order to design and interpret experiments in different circumstances. PMID- 20028468 TI - Glucose 1-phosphate is efficiently taken up by potato (Solanum tuberosum) tuber parenchyma cells and converted to reserve starch granules. AB - Reserve starch is an important plant product but the actual biosynthetic process is not yet fully understood. Potato (Solanum tuberosum) tuber discs from various transgenic plants were used to analyse the conversion of external sugars or sugar derivatives to starch. By using in vitro assays, a direct glucosyl transfer from glucose 1-phosphate to native starch granules as mediated by recombinant plastidial phosphorylase was analysed. Compared with labelled glucose, glucose 6 phosphate or sucrose, tuber discs converted externally supplied [(14)C]glucose 1 phosphate into starch at a much higher rate. Likewise, tuber discs from transgenic lines with a strongly reduced expression of cytosolic phosphoglucomutase, phosphorylase or transglucosidase converted glucose 1 phosphate to starch with the same or even an increased rate compared with the wild-type. Similar results were obtained with transgenic potato lines possessing a strongly reduced activity of both the cytosolic and the plastidial phosphoglucomutase. Starch labelling was, however, significantly diminished in transgenic lines, with a reduced concentration of the plastidial phosphorylase isozymes. Two distinct paths of reserve starch biosynthesis are proposed that explain, at a biochemical level, the phenotype of several transgenic plant lines. PMID- 20028470 TI - Heterotrimeric G proteins regulate reproductive trait plasticity in response to water availability. AB - Phenotypic plasticity is the ability of one genotype to display different phenotypes under different environmental conditions. Although variation for phenotypic plasticity has been documented in numerous species, little is known about the genetic mechanisms underlying phenotypic plasticity. Given their widespread roles in hormonal and environmental signaling, we examined whether genes which encode heterotrimeric G proteins are plasticity genes. We grew multiple alleles of heterotrimeric G-protein mutants, together with wild-type Arabidopsis thaliana, under different watering regimes to determine the contributions of G-protein genes to phenotypic plasticity for a number of developmental and reproduction-related traits. G-protein mutations did not affect significantly the amount of phenotypic variation within an environment for any trait, but did affect significantly the amount of phenotypic plasticity for certain traits. AGB1, which encodes the beta subunit of the heterotrimeric G protein in Arabidopsis, is a plasticity gene and regulates reproductive trait plasticity in response to water availability, resulting in increased fitness (defined as seed production) under drought stress. PMID- 20028469 TI - Plant sphingolipids: decoding the enigma of the Sphinx. AB - Sphingolipids are a ubiquitous class of lipids present in a variety of organisms including eukaryotes and bacteria. In the last two decades, research has focused on characterizing the individual species of this complex family of lipids, which has led to a new field of research called 'sphingolipidomics'. There are at least 500 (and perhaps thousands of) different molecular species of sphingolipids in cells, and in Arabidopsis alone it has been reported that there are at least 168 different sphingolipids. Plant sphingolipids can be divided into four classes: glycosyl inositol phosphoceramides (GIPCs), glycosylceramides, ceramides, and free long-chain bases (LCBs). Numerous enzymes involved in plant sphingolipid metabolism have now been cloned and characterized, and, in general, there is broad conservation in the way in which sphingolipids are metabolized in animals, yeast and plants. Here, we review the diversity of sphingolipids reported in the literature, some of the recent advances in our understanding of sphingolipid metabolism in plants, and the physiological roles that sphingolipids and sphingolipid metabolites play in plant physiology. PMID- 20028471 TI - Mapping quantitative trait loci associated with chilling requirement, heat requirement and bloom date in peach (Prunus persica). AB - *Chilling requirement, together with heat requirement, determines the bloom date, which has an impact on the climatic distribution of the genotypes of tree species. The molecular basis of floral bud chilling requirement is poorly understood, despite its importance to the adaptation and production of fruit trees. In addition, the genetic nature of heat requirement and the genetic interrelationships among chilling requirement, heat requirement and bloom date remain unclear. *A peach (Prunus persica) F(2) population of 378 genotypes developed from two genotypes with contrasting chilling requirements was used for linkage map construction and quantitative trait loci (QTL) mapping. The floral bud chilling and heat requirements of each genotype were evaluated over 2 yr and the bloom date was scored over 4 yr. *Twenty QTLs with additive effects were identified for three traits, including one major QTL for chilling requirement and two major QTLs for bloom date. The majority of QTLs colocalized with QTLs for other trait(s). In particular, one genomic region of 2 cM, pleiotropic for the three traits, overlapped with the sequenced peach EVG region. *This first report on the QTL mapping of floral bud chilling requirement will facilitate marker assisted breeding for low chilling requirement cultivars and the map-based cloning of genes controlling chilling requirement. The extensive colocalization of QTLs suggests that there may be one unified temperature sensing and action system regulating chilling requirement, heat requirement and bloom date together. PMID- 20028472 TI - Dating the origins of polyploidy events. AB - is a widespread speciation mechanism, particularly in plants. Estimating the time of origin of polyploid species is important for understanding issues such as gene loss and changes in regulation and expression among homoeologous copies that coexist in a single genome owing to polyploidy. Polyploid species can originate in various ways; the effects of mode of origin, genetic system, and sampling on estimates of the age of polyploid origin using distances between alleles of polyploids and their diploid progenitors, or between homoeologous loci in a polyploid genome, are explored. Even in the simplest cases, simulations confirm that different loci are expected to give very different estimates of the date of origin. The time of polyploid origin is at least as old as the time estimated from comparison of an allele sampled from the polyploid with the most closely related allele in the diploid progenitor. The polyploidy literature often does not make clear the longstanding observation that the divergence of homoeologous copies in an allopolyploid tracks the divergence of diploid species, not the origin of the polyploid. Estimating the date of origin of a polyploid is difficult, and in some circumstances impossible. Skepticism about dates of polyploid origins is clearly warranted. PMID- 20028473 TI - Making a functional diploid: from polysomic to disomic inheritance. AB - One little understood feature of polyploid speciation is the transition from polysomic to disomic inheritance, and much recent attention has focused on the role of pairing genes in this process. Using computer simulations we studied the effects of mutations, chromosomal inversions, chiasma, neofunctionalization, subfunctionalization and selection on the evolution of disomic inheritance in a polyploid over 10 000 generations. We show that: the evolution of pairing genes is not essential for the establishment of disomic inheritance, as genetic drift, coupled with a threshold for homologue pairing fidelity, is sufficient to explain the transition from polysomic to disomic inheritance; high rates of recombination increase the number of generations required for disomic inheritance to become established; both neofunctionalization and subfunctionalization speed up the transition to disomic inheritance. The data suggest that during polyploid species establishment, selection will favour reduced chiasma number and/or more focused distribution. The data also suggest a new role for subfunctionalization in that it can drive disomic inheritance. The evolution of subfunctionalization in genes across the genome will then act to maintain genes in syntenic blocks and may explain why such regions are so highly conserved. PMID- 20028474 TI - Gene flow and population admixture as the primary post-invasion processes in common ragweed (Ambrosia artemisiifolia) populations in France. AB - *An improved inference of the evolutionary history of invasive species may be achieved by analyzing the genetic variation and population differentiation of recently established populations and their ancestral (historical) populations. Employing this approach, we investigated the role of gene flow in the post invasion evolution of common ragweed (Ambrosia artemisiifolia). *Using eight microsatellite loci, we compared genetic diversity and structure among nine pairs of historical and recent populations in France. Historical populations were reconstructed from herbarium specimens dated from the late 19th to early 20th century, whereas recent populations were collected within the last 5 yr. *Recent populations showed greater allelic and genetic diversity than did historical populations. Recent populations exhibited a lower level of population differentiation, shorter genetic distances among populations and more weakly structured populations than did historical populations. *Our results suggest that currently invasive populations have arisen from active gene flow and the subsequent admixture of historical populations, incorporating new alleles from multiple introductions. PMID- 20028475 TI - Freeze-thaw-induced embolism in Pinus contorta: centrifuge experiments validate the 'thaw-expansion hypothesis' but conflict with ultrasonic emission data. AB - *The 'thaw-expansion hypothesis' postulates that xylem embolism is caused by the formation of gas bubbles on freezing and their expansion on thawing. We evaluated the hypothesis using centrifuge experiments and ultrasonic emission monitoring in Pinus contorta. *Stem samples were exposed to freeze-thaw cycles at varying xylem pressure (P) in a centrifuge before the percentage loss of hydraulic conductivity (PLC) was measured. Ultrasonic acoustic emissions were registered on samples exposed to freeze-thaw cycles in a temperature chamber. *Freeze-thaw exposure of samples spun at -3 MPa induced a PLC of 32% (one frost cycle) and 50% (two cycles). An increase in P to -0.5 MPa during freezing had no PLC effect, whereas increased P during thaw lowered PLC to 7%. Ultrasonic acoustic emissions were observed during freezing and thawing at -3 MPa, but not in air-dried or water saturated samples. A decrease in minimum temperature caused additional ultrasonic acoustic emissions, but had no effect on PLC. *The centrifuge experiments indicate that the 'thaw-expansion hypothesis' correctly describes the embolization process. Possible explanations for the increase in PLC on repeated frost cycles and for the ultrasonic acoustic emissions observed during freezing and with decreasing ice temperature are discussed. PMID- 20028476 TI - The Brassica juncea BjCdR15, an ortholog of Arabidopsis TGA3, is a regulator of cadmium uptake, transport and accumulation in shoots and confers cadmium tolerance in transgenic plants. AB - *A bZIP transcription factor from Brassica juncea (BjCdR15) was isolated by the cDNA-amplified fragment length polymorphism technique after cadmium treatment. Sequence analysis indicated high similarity between BjCdR15 and Arabidopsis TGA3. In Arabidopsis, TGA3 transcription is also induced by cadmium; hence, we investigated whether BjCdR15 is involved in cadmium tolerance and whether it can functionally replace TGA3 protein in Arabidopsis tga3-2 mutant plants. *BjCdR15 expression was detected mainly in the epidermis and vascular system of cadmium treated plants, and increased in roots and leaves after cadmium treatment. The overexpression of BjCdR15 in Arabidopsis and tobacco enhanced cadmium tolerance: overexpressing plants showed high cadmium accumulation in shoots. Conversely, Arabidopsis tga3-2 mutant plants showed high cadmium content in roots and inhibition of its transport to the shoot. *We demonstrated that BjCdR15 can functionally replace TGA3: in 35S::BjCdR15-tga3-2 plants, the long-distance transport of cadmium from root to shoot was restored and these plants showed an increased cadmium content in shoots compared with all other assays. In addition, BjCdR15/TGA3 regulated the synthesis of phytochelatin synthase and the expression of several metal transporters. *The results indicate that BjCdR15/TGA3 transcription factors play a crucial role in the regulation of cadmium uptake by roots and in its long-distance root to shoot transport. BjCdR15/TGA3 may thus be considered as useful candidates for potential biotechnological applications in the phytoextraction of cadmium from polluted soils. PMID- 20028477 TI - Senescence-induced loss in photosynthesis enhances cell wall beta-glucosidase activity. AB - A link between senescence-induced decline in photosynthesis and activity of beta glucosidase is examined in the leaves of Arabidopsis. The enzyme is purified and characterized. The molecular weight of the enzyme is 58 kDa. It shows maximum activity at pH 5.5 and at temperature of 50 degrees C. Photosynthetic measurements and activity of the enzyme are conducted at different developmental stages including senescence of leaves. Senescence causes a significant loss in total chlorophyll, stomatal conductance, rate of evaporation and in the ability of the leaves for carbon dioxide fixation. The process also brings about a decline in oxygen evolution, quantum yield of photosystem II (PS II) and quantum efficiency of PS II photochemistry of thylakoid membrane. The loss in photosynthesis is accompanied by a significant increase in the activity of the cell wall-bound beta-glucosidase that breaks down polysaccharides to soluble sugars. The loss in photosynthesis as a signal for the enhancement in the activity of the enzyme is confirmed from the observation that incubation of excised mature leaves in continuous dark or in light with a photosynthesis inhibitor 3-(3,4-dichlorophenyl)-1, 1-dimethylurea (DCMU) that leads to sugar starvation enhances the activity of the enzyme. The work suggests that in the background of photosynthetic decline, the polysaccharides bound to cell wall that remains intact even during late phase of senescence may be the last target of senescing leaves for a possible source of sugar for remobilization and completion of the energy-dependent senescence program. PMID- 20028478 TI - Reactive oxygen species in abiotic stress signaling. AB - Reactive oxygen species (ROS) are known to accumulate during abiotic stresses, and different cellular compartments respond to them by distinctive profiles of ROS formation. In contrast to earlier views, it is becoming increasingly evident that even during stress, ROS production is not necessarily a symptom of cellular dysfunction but might represent a necessary signal in adjusting the cellular machinery to the altered conditions. ROS can modulate many signal transduction pathways, such as mitogen-activated protein kinase cascades, and ultimately influence the activity of transcription factors. However, the picture of ROS mediated signaling is still fragmentary and the issues of ROS perception as well as the signaling specificity remain open. Here, we review some of the recent advances in plant abiotic stress signaling with emphasis on processes known to be affected heavily by ROS. PMID- 20028479 TI - In-depth analysis of the distinctive effects of norflurazon implies that tetrapyrrole biosynthesis, organellar gene expression and ABA cooperate in the GUN-type of plastid signalling. AB - Application of norflurazon (NF) damages plastids, induces photobleaching and represses expression of the nuclear LHCB1.2 gene encoding a light-harvesting protein. In genomes uncoupled (gun) mutants, LHCB1.2 expression is maintained in the presence of NF. The mutants gun2, gun4 and gun5 exhibit perturbations in tetrapyrrole biosynthesis, but gun1 is defective in organellar gene expression (OGE). How gun mutations affect nuclear gene expression (NGE) and why the signals elicited by the two types evoke the same response remains unknown. Here we show that the carotenoid biosynthesis inhibitors amitrole and flurochloridone can replace NF in gun assays, whereas novel tetrapyrrole pathway mutations do not provoke a gun phenotype. Changes in haem levels also do not account for LHCB1.2 derepression in NF-treated gun mutants. Pigment measurements indicated that gun mutants are not resistant to NF, but gun2, gun4 and gun5 retain low levels of lutein, as well as of neoxanthin and violaxanthin, the precursors of abscisic acid (ABA). This might explain the enhanced ABA sensitivity of gun4 and gun5 plants found in germination assays. Metabolite profiling and analyses of reactive oxygen species and cellular redox state failed to suggest a link between gun mutations and altered LHCB1.2 expression. However, in contrast to NF-treated wild type plants, gun mutants retain to a marked extent the capability to express the plastome-encoded proteins AtpB and RbcL. This, together with the finding that application of ABA can partially restore LHCB1.2 expression in NF-treated wild type plants, supports the view that tetrapyrrole, OGE and ABA signalling are interconnected. PMID- 20028480 TI - Is plant endophyte-mediated defensive mutualism the result of oxidative stress protection? AB - In this review, we discuss the biology and beneficial effects of plant endophytes on host plants. The current explanation of endophyte protection (defensive mutualism) of host plants is based on the secondary metabolites (alkaloids) with antiherbivore properties produced by the symbiotic association between host plant and endophytes. We propose an alternative explanation of the mechanism of host protection through enhanced stress tolerance to oxidative stress. Several studies have demonstrated the production of different compounds (phenolics) with antioxidant capacity in endophyte-infected plants. Endophytes may also produce mannitol, other carbohydrates and small molecules (proline) with antioxidant capacity. We suggest that enhanced antioxidant production by symbiotic plants may be the result of the production of reactive oxygen species (ROS) by endophytes. In turn, symbiotic plants are protected from oxidative stress produced by plant diseases, droughts, heavy metals and other oxidative stressors by the production of antioxidants. We also discuss the lichen symbiosis and evaluate whether management of ROS also plays a role in this defensive mutualism. Future experiments are needed to evaluate the hypothesis that antioxidants are responsible for enhanced stress tolerance in endophyte-infected plants. PMID- 20028481 TI - The role of reactive oxygen species in signalling from chloroplasts to the nucleus. AB - The coordination of chloroplast function with the rest of cellular activity requires a continual stream of communication from this organelle to the nucleus. Chloroplasts are major sites of the production of reactive oxygen species (ROS) as either by-products of the reduction of molecular oxygen (O(2)) or its excitation in the presence of highly energised pigments. Such ROS, while potentially damaging to the cell, are also important initiators or transducers of signals from these organelles to the nucleus in response to environmental cues. ROS can initiate such retrograde signalling pathways that trigger either programmed cell death or adjustment to changed conditions. Such different outcomes have implications for the way in which signal transduction by ROS is accomplished and is the subject of this review. In response to mild-stress situations, and as a consequence of their reactivity or because of their containment by cellular antioxidant systems, it is proposed that ROS engage with or initiate signalling at or very near their site of production. In contrast, under more extreme conditions, ROS are proposed to diffuse away from their site of production and consequently elicit a different set of signalling events. PMID- 20028482 TI - Fruit cuticle lipid composition during development in tomato ripening mutants. AB - Recent studies suggest that fruit cuticle is an important contributing factor to tomato (Solanum lycopersicum) fruit shelf life and storability. Moreover, it has been hypothesized that variation in fruit cuticle composition may underlie differences in traits such as fruit resistance to desiccation and microbial infection. To gain a better understanding of cuticle lipid composition diversity during fruit ontogeny and to assess if there are common features that correlate with ripening, we examined developmental changes in fruit cuticle wax and cutin monomer composition of delayed-ripening tomato fruit mutants, ripening inhibitor (rin) and non-ripening (nor) and delayed-ripening landrace Alcobaca. Previous reports show that fruit ripening processes such as climacteric ethylene production, cell wall degradation and color change are significantly delayed, or do not occur, in these lines. In the study presented here, however, we show that fruits from rin, nor and Alcobaca have cuticle lipid compositions that differ significantly from normal fruits of Ailsa Craig (AC) even at very early stages in fruit development, with continuing impacts throughout ripening. Moreover, rin, nor and the Alcobaca lines show quite different wax profiles from AC and each other throughout fruit development. Although cutin monomer composition differed much less than wax composition among the genotypes, all delayed-ripening lines possessed higher relative amounts of C(18) monomers than AC. Together, these results reveal new genetic associations between cuticle and fruit development processes and define valuable genetic resources to further explore the importance of cuticle in fruit shelf life. PMID- 20028484 TI - Prostaglandin E2 promotes Na1.8 trafficking via its intracellular RRR motif through the protein kinase A pathway. AB - Voltage-gated sodium channels (Na(v)) are essential for the initiation and propagation of action potentials in neurons. Na(v)1.8 activity is regulated by prostaglandin E(2) (PGE(2)). There is, however, no direct evidence showing the regulated trafficking of Na(v)1.8, and the molecular and cellular mechanism of PGE(2)-induced sodium channel trafficking is not clear. Here, we report that PGE(2) regulates the trafficking of Na(v)1.8 through the protein kinase A (PKA) signaling pathway, and an RRR motif in the first intracellular loop of Na(v)1.8 mediates this effect. In rat dorsal root ganglion (DRG) neurons, prolonged PGE(2) treatment enhanced Na(v)1.8 currents by increasing the channel density on the cell surface. Activation of PKA by forskolin had the same effect on DRG neurons and human embryonic kidney 293T cells expressing Na(v)1.8. Inhibition of PKA completely blocked the PGE(2)-promoted effect on Na(v)1.8. Mutation of five PKA phosphorylation sites or the RRR motif in the first intracellular loop of Na(v)1.8 abolished the PKA-promoted Na(v)1.8 surface expression. Furthermore, a membrane-tethered peptide containing the intracellular RRR motif disrupted the PGE(2)-induced promotion of the Na(v)1.8 current in DRG neurons. Our data indicate that PGE(2) promotes the surface expression of Na(v)1.8 via an intracellular RRR motif, and provide a novel mechanism for functional modulation of Na(v)1.8 by hyperalgesic agents. PMID- 20028483 TI - Expanding the definition of the classical bipartite nuclear localization signal. AB - Nuclear localization signals (NLSs) are amino acid sequences that target cargo proteins into the nucleus. Rigorous characterization of NLS motifs is essential to understanding and predicting pathways for nuclear import. The best characterized NLS is the classical NLS (cNLS), which is recognized by the cNLS receptor, importin-alpha. cNLSs are conventionally defined as having one (monopartite) or two clusters of basic amino acids separated by a 9-12 aa linker (bipartite). Motivated by the finding that Ty1 integrase, which contains an unconventional putative bipartite cNLS with a 29 aa linker, exploits the classical nuclear import machinery, we assessed the functional boundaries for linker length within a bipartite cNLS. We confirmed that the integrase cNLS is a bona fide bipartite cNLS, then carried out a systematic analysis of linker length in an obligate bipartite cNLS cargo, which revealed that some linkers longer than conventionally defined can function in nuclear import. Linker function is dependent on the sequence and likely the inherent flexibility of the linker. Subsequently, we interrogated the Saccharomyces cerevisiae proteome to identify cellular proteins containing putative long bipartite cNLSs. We experimentally confirmed that Rrp4 contains a bipartite cNLS with a 25 aa linker. Our studies show that the traditional definition of bipartite cNLSs is too restrictive and linker length can vary depending on amino acid composition. PMID- 20028485 TI - Rab10 regulates phagosome maturation and its overexpression rescues Mycobacterium containing phagosomes maturation. AB - Phagosome maturation follows a defined biochemical program and, in the vast majority of cases, the microbe inside the phagosome is killed and digested. Although, an important number of pathogens, including Mycobacterium tuberculosis, which kills around two million people every year, have acquired the ability to survive, and even replicate by arresting phagosomal maturation. To identify more of the machinery involved in phagocytosis and phagosomal maturation, we investigated the function of Rab10 in engulfment and maturation of inert particles and Mycobacterium bovis bacille Calmette-Guerin (BCG). We showed that Rab10 association with phagosomes is transient and confocal microscopy revealed detectible levels of Rab10 on phagosomal membranes at very early time-points, occurring even before Rab5 acquisition. Rab10 recruitment had strong functional consequence, as the knockdown of endogenous Rab10 by RNA interference or overexpression of Rab10 dominant-negative mutant delayed maturation of phagosomes of IgG-opsonized latex beads or heat killed-mycobacteria. These results can be explained, at least in part, by the involvement of Rab10 in recycling of some phagosomal components. More importantly, overexpression of the constitutively active mutant of Rab10 partially rescued live-Mycobacterium-containing phagosomes maturation. Indeed, we found that the membrane harbouring Mycobacterium acquired early endosome antigen 1 (EEA-1), a marker excluded from phagosomes in control cells. Altogether these results indicate that Rab10, acting upstream of Rab5, plays a prominent role in phagolysosome formation and can modulate Mycobacterium containing phagosomes maturation. PMID- 20028486 TI - Glucosylceramide biosynthesis is involved in Golgi morphology and protein secretion in plant cells. AB - Lipids have an established role as structural components of membranes or as signalling molecules, but their role as molecular actors in protein secretion is less clear. The complex sphingolipid glucosylceramide (GlcCer) is enriched in the plasma membrane and lipid microdomains of plant cells, but compared to animal and yeast cells, little is known about the role of GlcCer in plant physiology. We have investigated the influence of GlcCer biosynthesis by glucosylceramide synthase (GCS) on the efficiency of protein transport through the plant secretory pathway and on the maintenance of normal Golgi structure. We determined that GlcCer is synthesized at the beginning of the plant secretory pathway [mainly endoplasmic reticulum (ER)] and that D,L-threo-1-phenyl-2-decanoyl amino-3 morpholino-propanol (PDMP) is a potent inhibitor of plant GCS activity in vitro and in vivo. By an in vivo confocal microscopy approach in tobacco leaves infiltrated with PDMP, we showed that the decrease in GlcCer biosynthesis disturbed the transport of soluble and membrane secretory proteins to the cell surface, as these proteins were partly retained intracellularly in the ER and/or Golgi. Electron microscopic observations of Arabidopsis thaliana root cells after high-pressure freezing and freeze substitution evidenced strong morphological changes in the Golgi bodies, pointing to a link between decreased protein secretion and perturbations of Golgi structure following inhibition of GlcCer biosynthesis in plant cells. PMID- 20028488 TI - Psychometric properties of the revised Norwegian dispositional resilience (hardiness) scale. AB - In the 30 years that have elapsed since it was first introduced, the concept of hardiness has continued to attract the attention and interest of researchers from all over the world. The purpose of this study was to examine the reliability and factor structure of a revised Norwegian hardiness scale (Dispositional Resilience Scale 15; DRS-15). Using exploratory and confirmatory factor analytic strategies with a large sample of working adults (N = 7,280), support was found for a hierarchical structure comprising a general hardiness dimension and three sub dimensions (commitment, control, and challenge). Overall, the results support the reliability and validity of the revised DRS-15 and underscore the importance of examining the psychometric properties and cultural appropriateness of translated scales. PMID- 20028487 TI - Rab27a and Rab27b regulate neutrophil azurophilic granule exocytosis and NADPH oxidase activity by independent mechanisms. AB - Neutrophils rely on exocytosis to mobilize receptors and adhesion molecules and to release microbicidal factors. This process should be strictly regulated because uncontrolled release of toxic proteins would be injurious to the host. In vivo studies showed that the small GTPase Rab27a regulates azurophilic granule exocytosis. Using mouse neutrophils deficient in Rab27a (Rab27a(ash/ash)), Rab27b [Rab27b knockout (KO)] or both [Rab27a/b double KO (DoKo)], we investigated the role of the Rab27 isoforms in neutrophils. We found that both Rab27a and Rab27b deficiencies impaired azurophilic granule exocytosis. Rab27a(ash/ash) neutrophils showed upregulation of Rab27b expression which did not compensate for the secretory defects observed in Rab27a-deficient cells, suggesting that Rab27 isoforms play independent roles in neutrophil exocytosis. Total internal reflection fluorescence microscopy analysis showed that Rab27a(ash/ash) and Rab27b KO neutrophils have a decreased number of azurophilic granules near the plasma membrane. The effect was exacerbated in Rab27a/b DoKo neutrophils. Rab27 deficient neutrophils showed impaired activation of the reduced nicotinamide adenine dinucleotide phosphate (NADPH) oxidase at the plasma membrane although intraphagosomal reactive oxygen species (ROS) production was not affected. Exocytosis of secretory vesicles in Rab27-deficient neutrophils was functional, suggesting that Rab27 GTPases selectively control the exocytosis of neutrophil granules. PMID- 20028489 TI - Antibody-mediated rejection (AMR) after pancreas and pancreas-kidney transplantation. AB - Antibody-mediated rejection (AMR) requires specific diagnostic tools and treatment and is associated with lower graft survival. We prospectively screened C4d in pancreas (n = 35, in 27 patients) and kidney (n = 33, in 21 patients) for cause biopsies. Serum amylase and lipase, amylasuria, fasting blood glucose (FBG) and 2-h capillary glucose (CG) were also analysed. We found that 27.3% of kidney biopsies and 43% of pancreatic biopsies showed C4d staining (66.7% and 53.3% diffuse in peritubular and interacinar capillaries respectively). Isolated exocrine dysfunction was the main indication for pancreas biopsy (54.3%) and was followed by both exocrine and endocrine dysfunctions (37.1%) and isolated endocrine dysfunction (8.6%). Laboratorial parameters were comparable between T cell mediated rejection and AMR: amylase 151.5 vs. 149 U/l (P = 0.075), lipase 1120 vs. 1288.5 U/l (P = 0.83), amylasuria variation 46.5 vs. 61% (P = 0.97), FBG 69 vs. 97 mg/dl (P = 0.20) and 2-h CG maximum 149.5 vs. 197.5 mg/dl (P = 0.49) respectively. Amylasuria values after treatment correlated with pancreas allograft loss (P = 0.015). These data suggest that C4d staining should be routinely investigated when pancreas allograft dysfunction is present because of its high detection rate in cases of rejection. PMID- 20028490 TI - Pre-emptive antiviral therapy in living donor liver transplantation for hepatitis C: observation based on a single-center experience. AB - Reports of large series in living donor liver transplantation (LDLT) for hepatitis C virus infection (HCV) are scarce. Between 1996 and 2008, 105 LDLTs were performed at the University of Tokyo for HCV. Rapid induction of antiviral treatment with interferon (IFN) and ribavirin (RBV) was attempted per protocol regardless of the clinical presentation of recurrent HCV (pre-emptive treatment approach). Treatment was continued for 12 months after serum HCV-RNA became negative (ETR: end-of-treatment response) and judged as a sustained viral response (SVR) after another 6 months of negative results without treatment. A fixed treatment period was not defined unless an ETR was achieved (no-stopping approach). Flexible dose adjustments were allowed. Ninety-five patients were eligible for pre-emptive therapy. Forty-three (45%) patients experienced an ETR, and 32 (34%) achieved SVR. Nonadherence to full-dose INF and RBV had little impact on the viral response. Evaluation using the Kaplan-Meier method to incorporate the cumulative time-dependent nature of the no-stopping approach estimated SVR rate at 53% by the fifth year. Survival rate at 5 years was 79% for the HCV recipients and did not differ significantly from our non-HCV series. In LDLT for HCV, pre-emptive IFN-RBV-based treatment with the application of no stopping approach is feasible and effective. PMID- 20028491 TI - Bilateral urinary leak originating from the native ureters in a dual kidney transplant patient. PMID- 20028492 TI - A new composite midface allotransplantation model with sensory and motor reinnervation. AB - In this study, we extended application of face transplantation model in rat by incorporation of vascularized premaxilla, and nose with infraorbital and facial nerves for evaluation of allotransplanted sensory and motor nerve functional recovery. In group I (n = 3) the dissection technique is studied. In group II (n = 5) isotransplantations were performed. In group III (n = 5) allotransplantations were performed under Cyclosporin A monotherapy. Grafts; composed of nose, lower lip, and premaxilla; were dissected. Infraorbital nerve and facial nerve were included into the transplant. A heterotopic transplantation was performed to inguinal region of recipient. Nerve coaptations were performed between infraorbital-sapheneous nerve and facial-femoral nerve. CT scan, somatosensory-evoked potential testing (SSEP), motor-evoked potential testing (MEP), and microangiography were used for evaluation. All transplants survived indefinitely over 100 days. Microangiography showed preserved vascularization of the graft. Computed tomography revealed vital premaxillary bone segments. SSEP and MEP confirmed recovery of motor and sensory functions and latencies reached 67% of normal infraorbital nerve value and 70% of normal facial nerve value at 100 days post-transplant. We have introduced new midface transplant model of composite midface allograft with sensory and motor units. In this model, motor and sensory functional recovery was confirmed at 100 days post-transplant. PMID- 20028493 TI - Supporting evidence-based practice for nurses through information technologies. AB - PURPOSE: To evaluate the usability of mobile information terminals, such as personal digital assistants (PDAs) or Tablet personal computers, to improve access to information resources for nurses and to explore the relationship between PDA or Tablet-supported information resources and outcomes. BACKGROUND: The authors evaluated an initiative of the Nursing Secretariat, Ontario Ministry of Health and Long-Term Care, which provided nurses with PDAs and Tablet PCs, to enable Internet access to information resources. Nurses had access to drug and medical reference information, best practice guidelines (BPGs), and to abstracts of recent research studies. METHOD: The authors took place over a 12-month period. Diffusion of Innovation theory and the Promoting Action on Research Implementation in Health Services (PARIHS) model guided the selection of variables for study. A longitudinal design involving questionnaires was used to evaluate the impact of the mobile technologies on barriers to research utilization, perceived quality of care, and on nurses' job satisfaction. The setting was 29 acute care, long-term care, home care, and correctional organizations in Ontario, Canada. The sample consisted of 488 frontline-nurses. RESULTS: Nurses most frequently consulted drug and medical reference information, Google, and Nursing PLUS. Overall, nurses were most satisfied with the Registered Nurses Association of Ontario (RNAO) BPGs and rated the RNAO BPGs as the easiest resource to use. Among the PDA and Tablet users, there was a significant improvement in research awareness/values, and in communication of research. There was also, for the PDA users only, a significant improvement over time in perceived quality of care and job satisfaction, but primarily in long-term care settings. IMPLICATIONS: It is feasible to provide nurses with access to evidence based practice resources via mobile information technologies to reduce the barriers to research utilization. PMID- 20028494 TI - Fourteen-year survival of a renal graft reused 2 years after initial transplantation: a case report. AB - We report on the successful regrafting of a transplanted kidney. The donor kidney was first transplanted into a 32-year-old patient with renal atrophy. More than 2 years later, he suffered from severe grand mal seizure with brain edema and the patient met the criteria for brain death. The well-functioning graft was recovered and subsequently transplanted into a 66-year-old woman with chronic glomerular nephritis. Neither the first nor the second recipient experienced any acute rejection. To date, more than 14 years later, she is in good health with excellent graft function. This case report implies that excellent long-term graft function is viable in a graft reused 2 years after the initial transplantation. PMID- 20028495 TI - Successful living donor liver transplantation for severe hepatic GVHD histologically resembling autoimmune hepatitis after bone marrow transplantation from the same sibling donor. AB - A 30-year-old woman developed severe liver dysfunction 1 year after bone marrow transplantation (BMT) from an HLA-identical sibling donor for B lymphoblastic leukemia (B-ALL) during the tapering of cyclosporin A. The histologic picture resembled autoimmune hepatitis (AIH), although neither autoantibody nor hypergammaglobulinemia was detected. She entered hepatic coma, and underwent living donor liver transplantation from the same donor on day 421 after BMT. She is well 18 months after the procedure, showing normal liver function and hematopoiesis. AIH-like hepatic graft-versus-host disease (GVHD) has not been documented. This patient is the second case of living donor liver transplantation for hepatic GVHD from the same donor. PMID- 20028496 TI - Combined liver and kidney transplantation in acute intermittent porphyria. AB - We report two patients with acute intermittent porphyria (AIP) who were successfully treated with combined liver and kidney transplantation. Both had a very poor quality of life as a result of years of frequent acute porphyria symptoms, chronic peripheral neuropathy and renal failure requiring dialysis. After transplantation, clinical and biochemical signs of porphyria disappeared. The excretion pattern of porphyrin precursors normalized within the first day and plasma porphyrins returned to normal within a week. These and other recent cases have clarified previous concerns and have helped to formulate the indications for and the timing of transplantation in AIP. PMID- 20028497 TI - Lack of serologic immunity against vaccine-preventable diseases in children after thoracic transplantation. AB - We investigated whether children after heart- (HTx) or heart-lung transplantation (HLTx) show protective antibody levels against recommended vaccinations, whether vaccination schedules are completed and which factors influence serologic immunity. We performed a cross sectional ELISA - quantification of specific antibodies in 46 patients after pediatric thoracic Tx. Findings were correlated to vaccination history, age at Tx, clinical course and immunosuppressive regimen. We found protective antibody levels against diphtheria in 74% of patients, against tetanus in 22%, against Haemophilus influenzae type b in 30% and against Streptococcus pneumoniae in 59%. Antibody concentrations against live attenuated vaccines were significantly lower in children transplanted in the first 2 years of life. Antibodies were absent for measles in 55% of late - and 81% of early transplanted children, for mumps in 66%/94%, for rubella in 30%/56% and for Varicella in 34%/63%. We found significant correlation of low antibody concentrations and age at Tx. Patients without protective antibody concentrations had significantly longer use of steroids. Vaccination schedules were incomplete or delayed in the majority of patients associated with more days in hospital pre Tx. Our study shows that closer adherence to pretransplantation vaccination schedules and also post-transplantation monitoring of antibody levels are required in transplant patients. PMID- 20028498 TI - Knowledge, use and ecology of golden thistle (Scolymus hispanicus L.) in central Spain. AB - BACKGROUND: This paper assesses the current ethnobotanical knowledge, use and management of Scolymus hispanicus L. in two localities of Central Spain and the relation with its natural abundance. It also addresses the influence of sociodemographic factors such as age, gender and time living in the village in the variation of knowledge and practice levels. METHODS: During 2007 and 2008, 99 semi-structured interviews and a questionnaire were made to a random stratified sample by sex and age, asking them about their traditional knowledge and practices (use and gathering) of Scolymus hispanicus. A knowledge and practice (KP) index was created based on the answers to the questionnaire. RESULTS AND DISCUSSION: Scolymus hispanicus is still gathered and consumed by 20% and 35% of the informants, respectively. According to the KP index, the knowledge and practice level is similar in both villages. Age and time living in the village are the factors that better explain the variability in the KP level. People living for more than ten years in the village and those older than 60 years have the highest knowledge level, whereas the younger than 19 the lowest. CONCLUSIONS: Our data suggests that the prevalence of ethnobotanical knowledge and uses depends more on the cultural importance of the plant and the transmission of such popular knowledge than on the resource's abundance. PMID- 20028499 TI - Women convicted for violent offenses: adverse childhood experiences, low level of education and poor mental health. AB - BACKGROUND: In past years, the female offender population has grown, leading to an increased interest in the characteristics of female offenders. The aim of this study was to assess the prevalence of female violent offending in a Swiss offender population and to compare possible socio-demographic and offense-related gender differences. METHODS: Descriptive and bivariate logistic regression analyses were performed for a representative sample of N = 203 violent offenders convicted in Zurich, Switzerland. RESULTS: 7.9% (N = 16) of the sample were female. Significant gender differences were found: Female offenders were more likely to be married, less educated, to have suffered from adverse childhood experiences and to be in poor mental health. Female violent offending was less heterogeneous than male violent offending, in fact there were only three types of violent offenses females were convicted for in our sample: One third were convicted of murder, one third for arson and only one woman was convicted of a sex offense. CONCLUSIONS: The results of our study point toward a gender-specific theory of female offending, as well as toward the importance of developing models for explaining female criminal behavior, which need to be implemented in treatment plans and intervention strategies regarding female offenders. PMID- 20028501 TI - Thrombosis of abdominal aorta during cisplatin-based chemotherapy of testicular seminoma - a case report. AB - BACKGROUND: Vascular complications occurring during cisplatin-based chemotherapy of germ cell tumours are inadequately recognized to date. CASE PRESENTATION: A 49 year old man with advanced seminoma underwent two courses of chemotherapy according to the PEB regimen. Upon restaging, two thrombotic deposits were noted in the descending part of the thoracic aorta and in the infrarenal abdominal aorta, respectively. Although thrombotic plaques caused aortic occlusion of about 30%, no clinical signs of malperfusion of limbs were registered. The patient was placed on anticoagulant therapy. Six months after completion of chemotherapy, thrombotic deposits had completely resolved. In the absence of other predisposing factors, it must be assumed that cisplatin-based chemotherapy represented a strong stimulus for arterial thrombosis in the aorta. CONCLUSIONS: This is the first case of endo-aortic thrombosis during chemotherapy for testicular germ cell cancer. Providers of chemotherapy must be aware of arterial thrombosis even in young patients with testicular cancer. PMID- 20028500 TI - Co-infection by human immunodeficiency virus type 1 (HIV-1) and human T cell leukemia virus type 1 (HTLV-1): does immune activation lead to a faster progression to AIDS? AB - BACKGROUND: Recent data have shown that HTLV-1 is prevalent among HIV positive patients in Mozambique, although the impact of HTLV-1 infection on HIV disease progression remains controversial. Our aim was to determine the phenotypic profile of T lymphocytes subsets among Mozambican patients co-infected by HIV and HTLV-1. METHODS: We enrolled 29 patients co-infected by HTLV-1 and HIV (co infected), 59 patients mono-infected by HIV (HIV) and 16 healthy controls (HC), respectively.For phenotypic analysis, cells were stained with the following fluorochrome-labeled anti-human monoclonal antibodies CD4-APC, CD8-PerCP, CD25 PE, CD62L-FITC, CD45RA-FITC. CD45RO-PE, CD38-PE; being analysed by four-colour flow cytometry. RESULTS: We initially found that CD4+ T cell counts were significantly higher in co-infected, as compared to HIV groups. Moreover, CD4+ T Lymphocytes from co-infected patients presented significantly higher levels of CD45RO and CD25, but lower levels of CD45RA and CD62L, strongly indicating that CD4+ T cells are more activated under HTLV-1 plus HIV co-infection. CONCLUSION: Our data indicate that HTLV-1/HIV co-infected patients progress with higher CD4+ T cell counts and higher levels of activation markers. In this context, it is conceivable that in co-infected individuals, these higher levels of activation may account for a faster progression to AIDS. PMID- 20028502 TI - The presence of postmenopausal bleeding as prognostic parameter in patients with endometrial cancer: a retrospective multi-center study. AB - BACKGROUND: To date, there is no consensus on the utility of screening procedures for the early detection of endometrial cancer. The value of transvaginal ultrasound for screening of asymptomatic endometrial cancer has been discussed controversially. This study was conducted to evaluate whether asymptomatic patients with endometrial cancer have a better prognosis than symptomatic patients with endometrial cancer diagnosed after postmenopausal bleeding. METHODS: In the present multi-center study, the effect of the presence of postmenopausal bleeding on prognosis was evaluated retrospectively in 605 patients with endometrial cancer using patients' files. 543 patients (133 patients were asymptomatic, 410 patients were symptomatic) with endometrioid endometrial cancer were enrolled in all further analysis. Student's t-test, Cox regression analysis and Kaplan-Meier analysis were used were appropriate. RESULTS: Presence/absence of a postmenopausal bleeding was not associated with tumor stage (p = 0.2) and age at diagnosis (p = 0.5). Asymptomatic patients with endometrial cancer had a significantly higher rate of well and moderate differentiated tumors compared to symptomatic patients (p = 0.008). In univariable and multivariable survival analysis, tumor stage, tumor grade, and patients' age at diagnosis, but not presence/absence of a postmenopausal bleeding, were associated with disease free and overall survival. CONCLUSION: Asymptomatic patients with endometrial cancer have a higher rate of well differentiated tumors compared to patients with a postmenopausal bleeding prior to diagnosis. The prognosis of both groups of patients was similar. PMID- 20028503 TI - Prognostic impact of H3K27me3 expression on locoregional progression after chemoradiotherapy in esophageal squamous cell carcinoma. AB - BACKGROUND: Trimethylation of lysine 27 on histone H3 (H3K27me3) by enhancer of zeste homolog 2 (EZH2) is an epigenetic mark that mediates gene silencing. EZH2 is overexpressed and correlates with poor prognosis in many cancers. However, the clinical implication of H3K27me3 in human malignancies has not been well established. We wished to ascertain whether a correlation exists between the expression of H3K27me3 and clinical outcome in a group of patients with esophageal squamous cell carcinoma (ESCC) treated with definitive chemoradiotherapy (CRT). METHODS: The method of immunohistochemistry (IHC) was utilized to examine the protein expression of H3K27me3 in 98 pretreatment biopsy specimens of ESCC and in 30 samples of normal esophageal mucosa. The clinical/prognostic significance of H3K27me3 expression was statistically analyzed. RESULTS: The expression frequency and expression levels of H3K27me3 were significantly higher in ESCCs than in normal tissues. There was a positive correlation between H3K27me3 expression and WHO grade (P = 0.016), tumor size (P = 0.019), T status (P = 0.024), locoregional progression (P = 0.009) and EZH2 expression (P = 0.036). High H3K27me3 expression was associated with poor locoregional progression-free survival (LPFS) (P = 0.010) in ESCC. Further analysis demonstrated that H3K27me3 could stratify patient outcome in T2-3 (P = 0.048), N0 (P = 0.005) and M0 (P = 0.018) stages as well as in CRT effective group (P = 0.022). CONCLUSIONS: Our data suggests that H3K27me3 expression examined by IHC might be useful for stratifying LPFS for different subsets of ESCC patients treated with definitive CRT. PMID- 20028504 TI - GPs' views on transfer of information about terminally ill patients to the out-of hours co-operative. AB - BACKGROUND: In the Netherlands, the increase in of out-of-hours care that is provided by GP co-operatives is challenging the continuity of care for the terminally ill in general practice. Aim of this study is to investigate the views of general practitioners (GPs) on the transfer of information about terminally ill patients to the GP co-operatives. GPs were asked to give their view from two different perspectives: as a GP in their daily practice and as a locum in the GP co-operative. METHODS: Retrospective web based questionnaire sent to all 424 GPs in the Amsterdam region. RESULTS: With a response rate of 42%, 177 physicians completed the questionnaire. Transfer of information to the GP co-operative about most of their terminally ill patients was reported by 82% of the GPs and 5% did not do so for any of their patients. A faster than foreseen deterioration of the patient's situation was the most frequently reported reason for not transferring information. Of those who transferred information to the GP co-operative, more than 95% reported that they provided information about the diagnosis and terminally ill status of the patient. Information about medication, patient wishes regarding treatment, and prognosis was reported by respectively 90%, 87%, and 74% of the GPs. Less than 50% of the GPs reported that they transferred information about the patient's awareness of both the diagnosis and the prognosis, about the psychosocial context, and intolerances. In their role as locum, over 90% of the GPs wanted to receive information about the diagnosis, the terminally ill status of the patient, the medication and the patient's wishes regarding treatment. CONCLUSIONS: Although most GPs reported that they transferred information about their terminally ill patients to the GP co operative, the content of this information varies considerably. Only 21% of the GPs, working out of hours as a locum, were satisfied with the quality of the information transferred. PMID- 20028505 TI - Functional diversity of human protein kinase splice variants marks significant expansion of human kinome. AB - BACKGROUND: Protein kinases are involved in diverse spectrum of cellular processes. Availability of draft version of the human genomic data in the year 2001 enabled recognition of repertoire of protein kinases. However, over the years the human genomic data is being refined and the current release of human genomic data has helped us to recognize a larger repertoire of over 900 human protein kinases represented mainly by splice variants. RESULTS: Many of these identified protein kinases are alternatively spliced products. Interestingly, some of the human kinase splice variants appear to be significantly diverged in terms of their functional properties as represented by incorporation or absence of one or more domains. Many sets of protein kinase splice variants have substantially different domain organization and in a few sets of splice variants kinase domains belong to different subfamilies of kinases suggesting potential participation in different signal transduction pathways. CONCLUSIONS: Addition or deletion of a domain between splice variants of multi-domain kinases appears to be a means of generating differences in the functional features of otherwise similar kinases. It is intriguing that marked sequence diversity within the catalytic regions of some of the splice variant kinases result in kinases belonging to different subfamilies. These human kinase splice variants with different functions might contribute to diversity of eukaryotic cellular signaling. PMID- 20028506 TI - Depression and post-traumatic stress disorder among Haitian immigrant students: implications for access to mental health services and educational programming. AB - BACKGROUND: Previous studies of Haitian immigrant and refugee youth have emphasized "externalizing" behaviors, such as substance use, high risk sexual behavior, and delinquency, with very little information available on "internalizing" symptoms, such as depression and anxiety. Analyzing stressors and "internalizing" symptoms offers a more balanced picture of the type of social and mental health services that may be needed for this population. The present study aims to: 1) estimate the prevalence of depression and post-traumatic stress disorder (PTSD) among Haitian immigrant students; and 2) examine factors associated with depression and PTSD to identify potential areas of intervention that may enhance psychosocial health outcomes among immigrant youth from Haiti in the U.S. METHODS: A stratified random sample of Haitian immigrant students enrolled in Boston public high schools was selected for participation; 84% agreed to be interviewed with a standardized questionnaire. Diagnosis of depression and PTSD was ascertained using the best estimate diagnosis method. RESULTS: The prevalence estimates of depression and PTSD were 14.0% and 11.6%; 7.9% suffered from comorbid PTSD and depression. Multivariate logistic regression demonstrated factors most strongly associated with depression (history of father's death, self report of schoolwork not going well, not spending time with friends) and PTSD (concern for physical safety, having many arguments with parents, history of physical abuse, and lack of safety of neighborhood). CONCLUSIONS: A significant level of depression and PTSD was observed. Stressors subsequent to immigration, such as living in an unsafe neighborhood and concern for physical safety, were associated with an increased risk of PTSD and should be considered when developing programs to assist this population. Reducing exposure to these stressors and enhancing access to social support and appropriate school-based and mental health services may improve educational attainment and psychosocial health outcomes among Haitian immigrant youth. PMID- 20028507 TI - Seropositivity of hepatitis B virus and hepatitis C virus dual infection among blood donors in Nyala teaching hospital. AB - The aim of this study was to determine the seropositivity of Hepatitis B virus (HBV) and Hepatitis C virus (HCV) dual infection among blood donors in Nyala Teaching Hospital, which is the biggest (400 beds) hospital in great Dar Fur of Western Sudan. 400 blood donors were tested serologically for the detection of HBsAg and anti-HCV antibodies. Only one (0.25%) out of the 400 examined blood donors was detected reactive for both HBsAg and anti-HCV antibodies. The study concluded that the seropositivity of HBV and HCV dual infection among population studied is uncommon. PMID- 20028508 TI - Residential traffic exposure and pregnancy-related outcomes: a prospective birth cohort study. AB - BACKGROUND: The effects of ambient air pollution on pregnancy outcomes are under debate. Previous studies have used different air pollution exposure assessment methods. The considerable traffic-related intra-urban spatial variation needs to be considered in exposure assessment. Residential proximity to traffic is a proxy for traffic-related exposures that takes into account within-city contrasts. METHODS: We investigated the association between residential proximity to traffic and various birth and pregnancy outcomes in 7,339 pregnant women and their children participating in a population-based cohort study. Residential proximity to traffic was defined as 1) distance-weighted traffic density in a 150 meter radius, and 2) proximity to a major road. We estimated associations of these exposures with birth weight, and with the risks of preterm birth and small size for gestational age at birth. Additionally, we examined associations with pregnancy-induced hypertension, (pre)eclampsia, and gestational diabetes. RESULTS: There was considerable variation in distance-weighted traffic density. Almost fifteen percent of the participants lived within 50 m of a major road. Residential proximity to traffic was not associated with birth and pregnancy outcomes in the main analysis and in various sensitivity analyses. CONCLUSIONS: Mothers exposed to residential traffic had no higher risk of adverse birth outcomes or pregnancy complications in this study. Future studies may be refined by taking both temporal and spatial variation in air pollution exposure into account. PMID- 20028509 TI - The role of virulence factors in the outcome of staphylococcal peritonitis in CAPD patients. AB - BACKGROUND: Peritonitis continues to be the most frequent cause of peritoneal dialysis (PD) failure, with an important impact on patient mortality. Gram positive cocci such as Staphylococcus epidermidis, other coagulase-negative staphylococci (CoNS), and Staphylococcus aureus are the most frequent etiological agents of PD-associated peritonitis worldwide. The objective of the present study was to compare peritonitis caused by S. aureus and CoNS and to evaluate the factors influencing outcome. METHODS: Records of 86 new episodes of staphylococcal peritonitis that occurred between 1996 and 2000 in the Dialysis unit of a single university hospital were studied (35 due to S. aureus, 24 to S. epidermidis and 27 to other CoNS). The production of slime, lipase, lecithinase, nuclease (DNAse), thermonuclease (TNAse), alpha- and beta-hemolysin, enterotoxins (SEA, SEB, SEC, SED) and toxic shock syndrome toxin-1 (TSST-1) was studied in S. aureus and CoNS. Antimicrobial susceptibility was evaluated based on the minimal inhibitory concentration determined by the E-test. Outcome predictors were evaluated by two logistic regression models. RESULTS: The oxacillin susceptibility rate was 85.7% for S. aureus, 41.6% for S. epidermidis, and 51.8% for other CoNS (p = 0.001). Production of toxins and enzymes, except for enterotoxin A and alpha-hemolysin, was associated with S. aureus episodes (p < 0.001), whereas slime production was positive in 23.5% of CoNS and 8.6% of S. aureus strains (p = 0.0047). The first model did not include enzymes and toxins due to their association with S. aureus. The odds of resolution were 9.5 times higher for S. epidermidis than for S. aureus (p = 0.02) episodes, and were similar for S. epidermidis and other CoNS (p = 0.8). The resolution odds were 68 times higher for non-slime producers (p = 0.001) and were not influenced by oxacillin resistance among vancomycin-treated cases (p = 0.89). In the second model, the resolution rate was similar for S. aureus and S. epidermidis (p = 0.70), and slime (p = 0.001) and alpha-hemolysin (p = 0.04) production were independent predictors of non-resolution. CONCLUSION: Bacterial species and virulence factors rather than antibiotic resistance influence the outcome of staphylococcal peritonitis. PMID- 20028510 TI - Survivin gene levels in the peripheral blood of patients with gastric cancer independently predict survival. AB - BACKGROUND: The detection of circulating tumor cells (CTC) is considered a promising tool for improving risk stratification in patients with solid tumors. We investigated on whether the expression of CTC related genes adds any prognostic power to the TNM staging system in patients with gastric carcinoma. METHODS: Seventy patients with TNM stage I to IV gastric carcinoma were retrospectively enrolled. Peripheral blood samples were tested by means of quantitative real time PCR (qrtPCR) for the expression of four CTC related genes: carcinoembryonic antigen (CEA), cytokeratin-19 (CK19), vascular endothelial growth factor (VEGF) and Survivin (BIRC5). RESULTS: Gene expression of Survivin, CK19, CEA and VEGF was higher than in normal controls in 98.6%, 97.1%, 42.9% and 38.6% of cases, respectively, suggesting a potential diagnostic value of both Survivin and CK19. At multivariable survival analysis, TNM staging and Survivin mRNA levels were retained as independent prognostic factors, demonstrating that Survivin expression in the peripheral blood adds prognostic information to the TNM system. In contrast with previously published data, the transcript abundance of CEA, CK19 and VEGF was not associated with patients' clinical outcome. CONCLUSIONS: Gene expression levels of Survivin add significant prognostic value to the current TNM staging system. The validation of these findings in larger prospective and multicentric series might lead to the implementation of this biomarker in the routine clinical setting in order to optimize risk stratification and ultimately personalize the therapeutic management of these patients. PMID- 20028511 TI - HES 130/0.4 impairs haemostasis and stimulates pro-inflammatory blood platelet function. AB - INTRODUCTION: Hydroxyethyl starch (HES) solutions are widely used for volume replacement therapy but are also known to compromise coagulation, impair renal function and increase long-term mortality. To test the hypotheses that HES 130/0.4 has fewer adverse effects than HES 200/0.5 and exerts anti-inflammatory properties, we compared the effects of HES 130/0.4, HES 200/0.5 and saline on in vitro haemostasis and pro-inflammatory platelet function. METHODS: Whole blood samples from healthy volunteers were mixed with 6% HES 130/0.4, 10% HES 200/0.5, or normal saline to achieve a final haemodilution rate of 10% or 40%. Haemostatic capacity was characterised by thromboelastography (ROTEM) and measurement for FXIIIa activity. Platelet activation and pro-inflammatory platelet functions were characterised by flow cytometry measuring the platelet activation marker CD62P and binding of fibrinogen to platelets as well as the formation of heterotypic platelet-leukocyte conjugates. RESULTS: Compared with saline, HES 130/0.4 dose dependently impaired formation and firmness of the fibrin clot but did not affect the fibrin crosslinking activity of FXIIIa. At 40% but not at 10% haemodilution rate, HES 200/0.5 also increased platelet fibrinogen binding and both HES solutions increased expression of CD62P, the main receptor for platelet-leukocyte adhesion. HES 130/0.4 but not HES 200/0.5 increased formation of platelet neutrophil conjugates and, to a lesser degree, platelet-monocyte conjugates. CONCLUSIONS: Our data demonstrate that HES 130/0.4 has similar adverse effects as HES 200/0.5. In particular, both types of HES impair coagulation capacity and stimulate, rather than attenuate, pro-inflammatory platelet function. PMID- 20028512 TI - Amyloid and tau cerebrospinal fluid biomarkers in HIV infection. AB - BACKGROUND: Because of the emerging intersections of HIV infection and Alzheimer's disease, we examined cerebrospinal fluid (CSF) biomarkers related of amyloid and tau metabolism in HIV-infected patients. METHODS: In this cross sectional study we measured soluble amyloid precursor proteins alpha and beta (sAPPalpha and sAPPbeta), amyloid beta fragment 1-42 (Abeta1-42), and total and hyperphosphorylated tau (t-tau and p-tau) in CSF of 86 HIV-infected (HIV+) subjects, including 21 with AIDS dementia complex (ADC), 25 with central nervous system (CNS) opportunistic infections and 40 without neurological symptoms and signs. We also measured these CSF biomarkers in 64 uninfected (HIV-) subjects, including 21 with Alzheimer's disease, and both younger and older controls without neurological disease. RESULTS: CSF sAPPalpha and sAPPbeta concentrations were highly correlated and reduced in patients with ADC and opportunistic infections compared to the other groups. The opportunistic infection group but not the ADC patients had lower CSF Abeta1-42 in comparison to the other HIV+ subjects. CSF t-tau levels were high in some ADC patients, but did not differ significantly from the HIV+ neuroasymptomatic group, while CSF p-tau was not increased in any of the HIV+ groups. Together, CSF amyloid and tau markers segregated the ADC patients from both HIV+ and HIV- neuroasymptomatics and from Alzheimer's disease patients, but not from those with opportunistic infections. CONCLUSIONS: Parallel reductions of CSF sAPPalpha and sAPPbeta in ADC and CNS opportunistic infections suggest an effect of CNS immune activation or inflammation on neuronal amyloid synthesis or processing. Elevation of CSF t-tau in some ADC and CNS infection patients without concomitant increase in p-tau indicates neural injury without preferential accumulation of hyperphosphorylated tau as found in Alzheimer's disease. These biomarker changes define pathogenetic pathways to brain injury in ADC that differ from those of Alzheimer's disease. PMID- 20028513 TI - Overdiagnosis in organised mammography screening in Denmark. A comparative study. AB - BACKGROUND: Overdiagnosis in cancer screening is the detection of cancer lesions that would otherwise not have been detected. It is arguably the most important harm. We quantified overdiagnosis in the Danish mammography screening programme, which is uniquely suited for this purpose, as only 20% of the Danish population has been offered organised mammography screening over a long time-period. METHODS: We collected incidence rates of carcinoma in situ and invasive breast cancer in areas with and without screening over 13 years with screening (1991 2003), and 20 years before its introduction (1971-1990). We explored the incidence increase comparing unadjusted incidence rates and used Poisson regression analysis to compensate for the background incidence trend, variation in age distribution and geographical variation in incidence. RESULTS: For the screened age group, 50 to 69 years, we found an overdiagnosis of 35% when we compared unadjusted incidence rates for the screened and non-screened areas, but after compensating for a small decline in incidence in older, previously screened women. Our adjusted Poisson regression analysis indicated a relative risk of 1.40 (95% CI: 1.35-1.45) for the whole screening period, and a potential compensatory drop in older women of 0.90 (95% CI: 0.88-0.96), yielding an overdiagnosis of 33%, which we consider the most reliable estimate. The drop in previously screened women was only present in one of the two screened regions and was small in absolute numbers. DISCUSSION: One in four breast cancers diagnosed in the screened age group in the Danish screening programme is overdiagnosed. Our estimate for Denmark is lower than that for comparable countries, likely because of lower uptake, lower recall rates and lower detection rates of carcinoma in situ. PMID- 20028514 TI - Identification of MarvelD3 as a tight junction-associated transmembrane protein of the occludin family. AB - BACKGROUND: Tight junctions are an intercellular adhesion complex of epithelial and endothelial cells, and form a paracellular barrier that restricts the diffusion of solutes on the basis of size and charge. Tight junctions are formed by multiprotein complexes containing cytosolic and transmembrane proteins. How these components work together to form functional tight junctions is still not well understood and will require a complete understanding of the molecular composition of the junction. RESULTS: Here we identify a new transmembrane component of tight junctions: MarvelD3, a four-span transmembrane protein. Its predicted transmembrane helices form a Marvel (MAL and related proteins for vesicle traffic and membrane link) domain, a structural motif originally discovered in proteins involved in membrane apposition and fusion events, such as the tight junction proteins occludin and tricellulin. In mammals, MarvelD3 is expressed as two alternatively spliced isoforms. Both isoforms exhibit a broad tissue distribution and are expressed by different types of epithelial as well as endothelial cells. MarvelD3 co-localises with occludin at tight junctions in intestinal and corneal epithelial cells. RNA interference experiments in Caco-2 cells indicate that normal MarvelD3 expression is not required for the formation of functional tight junctions but depletion results in monolayers with increased transepithelial electrical resistance. CONCLUSIONS: Our data indicate that MarvelD3 is a third member of the tight junction-associated occludin family of transmembrane proteins. Similar to occludin, normal expression of MarvelD3 is not essential for the formation of functional tight junctions. However, MarvelD3 functions as a determinant of epithelial paracellular permeability properties. PMID- 20028515 TI - A response to information criterion-based clustering with order-restricted candidate profiles in short time-course microarray experiments. AB - BACKGROUND: For gene expression data obtained from a time-course microarray experiment, Liu et al. developed a new algorithm for clustering genes with similar expression profiles over time. Performance of their proposal was compared with three other methods including the order-restricted inference based methodology of Peddada et al. In this note we point out several inaccuracies in Liu et al. and conclude that the order-restricted inference based methodology of Peddada et al. (programmed in the software ORIOGEN) indeed operates at the desired nominal Type 1 error level, an important feature of a statistical decision rule, while being computationally substantially faster than indicated by Liu et al. RESULTS: Application of ORIOGEN to the well-known breast cancer cell line data of Lobenhofer et al. revealed that ORIOGEN software took only 21 minutes to run (using 100,000 bootstraps with p = 0.0025), substantially faster than the 72 hours found by Liu et al. using Matlab. Also, based on a data simulated according to the model and parameters of simulation 1 (sigma2 = 1, M = 5) in [1] we found that ORIOGEN took less than 30 seconds to run in stark contrast to Liu et al. who reported that their implementation of the same algorithm in R took 2979.29 seconds. Furthermore, for the simulation studies reported in [1], unlike the claims made by Liu et al., ORIOGEN always maintained the desired false positive rate. According to Figure three in Liu et al. their algorithm had a false positive rate ranging approximately from 0.20 to 0.70 for the scenarios that they simulated. CONCLUSIONS: Our comparisons of run times indicate that the implementations of ORIOGEN's algorithm in Matlab and R by Liu et al. is inefficient compared to the publicly available JAVA implementation. Our results on the false positive rate of ORIOGEN suggest some error in Figure three of Liu et al., perhaps due to a programming error. PMID- 20028518 TI - A modeling and simulation study of siderophore mediated antagonism in dual species biofilms. AB - BACKGROUND: Several bacterial species possess chelation mechanisms that allow them to scavenge iron from the environment under conditions of limitation. To this end they produce siderophores that bind the iron and make it available to the cells later on, while rendering it unavailable to other organisms. The phenomenon of siderophore mediated antagonism has been studied to some extent for suspended populations where it was found that the chelation ability provides a growth advantage over species that do not have this possibility. However, most bacteria live in biofilm communities. In particular Pseudomonas fluorescens and Pseudomonas putida, the species that have been used in most experimental studies of the phenomenon, are known to be prolific biofilm formers, but only very few experimental studies of iron chelation have been published to date for the biofilm setting. We address this question in the present study. METHODS: Based on a previously introduced model of iron chelation and an existing model of biofilm growth we formulate a model for iron chelation and competition in dual species biofilms. This leads to a highly nonlinear system of partial differential equations which is studied in computer simulation experiments. CONCLUSIONS: (i) Siderophore production can give a growth advantage also in the biofilm setting, (ii) diffusion facilitates and emphasizes this growth advantage, (iii) the magnitude of the growth advantage can also depend on the initial inoculation of the substratum, (iv) a new mass transfer boundary condition was derived that allows to a priori control the expect the expected average thickness of the biofilm in terms of the model parameters. PMID- 20028517 TI - CXCR4 expression heterogeneity in neuroblastoma cells due to ligand-independent regulation. AB - BACKGROUND: CXCR4, the receptor for the chemokine stromal-derived factor 1 (SDF 1), has been shown to mediate many of the processes essential for cancer progression such as tumor cell proliferation, metastasis, and angiogenesis. To understand the role of CXCR4 in the biology of neuroblastoma, a disease that presents with wide spread metastases in over 50% of patients, we screened ten patient derived-neuroblastoma cell-lines for basal CXCR4 expression and sought to identify characteristics that correlate with tumor cell phenotype. RESULTS: All cell lines expressed CXCR4 mRNA at variable levels, that correlated well with three distinct classes of CXCR4 surface expression (low, moderate, or high) as defined by flow cytometry. Analysis of the kinetics of CXCR4 surface expression on moderate and high expressing cell lines showed a time-dependent down regulation of the receptor that directly correlated with cell confluency, and was independent of SDF1. Cell lysates showed the presence of multiple CXCR4 isoforms with three major species of approximately 87, 67 and 55 kDa associating with high surface expression, and two distinct species of 45 and 38 kDa correlating with low to null surface expression. Western blot analysis of CXCR4 immunoprecipitates showed that the 87 and 67 kDa forms were ubiquitinated, while the others were not. Finally, treatment of cells with a proteasome inhibitor resulted in down regulation of CXCR4 surface expression. CONCLUSIONS: Taken together, these data show that regulation of CXCR4 surface expression in neuroblastoma cells can occur independently of SDF-1 contribution arguing against an autocrine mechanism. Additionally these data suggest that post-translational modifications of CXCR4, in part through direct ubiquitination, can influence trafficking of CXCR4 to the surface of neuroblastoma cells in a ligand-independent manner. PMID- 20028516 TI - Identification and characterization of endonuclein binding proteins: evidence of modulatory effects on signal transduction and chaperone activity. AB - BACKGROUND: We have previously identified endonuclein as a cell cycle regulated WD-repeat protein that is up-regulated in adenocarcinoma of the pancreas. Now, we aim to investigate its biomedical functions. RESULTS: Using the cDNA encoding human endonuclein, we have expressed and purified the recombinant protein from Escherichia coli using metal affinity chromatography. The recombinant protein was immobilized to a column and by affinity chromatography several interacting proteins were purified from several litres of placenta tissue extract. After chromatography the eluted proteins were further separated by two-dimensional gel electrophoresis and identified by tandem mass spectrometry. The interacting proteins were identified as; Tax interaction protein 1 (TIP-1), Aalpha fibrinogen transcription factor (P16/SSBP1), immunoglobulin heavy chain binding protein (BiP), human ER-associated DNAJ (HEDJ/DNAJB11), endonuclein interaction protein 8 (EIP-8), and pregnancy specific beta-1 glycoproteins (PSGs). Surface plasmon resonance analysis and confocal fluorescence microscopy were used to further characterize the interactions. CONCLUSIONS: Our results demonstrate that endonuclein interacts with several proteins indicating a broad function including signal transduction and chaperone activity. PMID- 20028519 TI - Genetically-increased taste cell population with G(alpha)-gustducin-coupled sweet receptors is associated with increase of gurmarin-sensitive taste nerve fibers in mice. AB - BACKGROUND: The peptide gurmarin is a selective sweet response inhibitor for rodents. In mice, gurmarin sensitivity differs among strains with gurmarin sensitive C57BL and gurmarin-poorly-sensitive BALB strains. In C57BL mice, sweet responsive fibers of the chorda tympani (CT) nerve can be divided into two distinct populations, gurmarin-sensitive (GS) and gurmarin-insensitive (GI) types, suggesting the existence of two distinct reception pathways for sweet taste responses. By using the dpa congenic strain (dpa CG) whose genetic background is identical to BALB except that the gene(s) controlling gurmarin sensitivity are derived from C57BL, we previously found that genetically-elevated gurmarin sensitivity in dpa CG mice, confirmed by using behavioral response and whole CT nerve response analyses, was linked to a greater taste cell population co-expressing sweet taste receptors and a G(alpha)- protein, G(alpha)--gustducin. However, the formation of neural pathways from the increased taste cell population to nerve fibers has not yet been examined. RESULTS: Here, we investigated whether the increased taste cell population with G(alpha)--gustducin coupled sweet receptors would be associated with selective increment of GS fiber population or nonselective shift of gurmarin sensitivities of overall sweet responsive fibers by examining the classification of GS and GI fiber types in dpa CG and BALB mice. The results indicated that dpa CG, like C57BL, possess two distinct populations of GS and GI types of sweet-responsive fibers with almost identical sizes (dpa CG: 13 GS and 16 GI fibers; C57BL: 16 GS and 14 GI fibers). In contrast, BALB has only 3 GS fibers but 18 GI fibers. These data indicate a marked increase of the GS population in dpa CG. CONCLUSION: These results suggest that the increased cell population expressing T1r2/T1r3/G(alpha)--gustducin in dpa CG mice may be associated with an increase of their matched GS type fibers, and may form the distinct GS sweet reception pathway in mice. G(alpha)--gustducin may be involved in the GS sweet reception pathway and may be a key molecule for links between sweet taste receptors and cell type-specific-innervation by their matched fiber class. PMID- 20028520 TI - Acute hepatitis in three patients with systemic juvenile idiopathic arthritis taking interleukin-1 receptor antagonist. AB - PURPOSE: We investigated the etiology of acute hepatitis in three children with systemic Juvenile Idiopathic Arthritis (sJIA) taking Interleukin-1 receptor antagonist (IL1RA). METHODS: Laboratory and clinical data for three children with sJIA diagnosed at ages 13 months to 8 years who developed acute hepatitis during treatment with IL1RA were reviewed for evidence of sJIA flare, infection, macrophage activation syndrome (MAS), malignancy, and drug reaction. RESULTS: In all patients, hepatitis persisted despite cessation of known hepatotoxic drugs and in absence of known infectious triggers, until discontinuation of IL1RA. Liver biopsies had mixed inflammatory infiltrates with associated hepatocellular injury suggestive of an exogenous trigger. At the time of hepatitis, laboratory data and liver biopsies were not characteristic of MAS. In two patients, transaminitis resolved within one week of discontinuing IL1RA, the third improved dramatically in one month. CONCLUSIONS: Although sJIA symptoms improved significantly on IL1RA, it appeared that IL1RA contributed to the development of acute hepatitis. Hepatitis possibly occurred as a result of an altered immune response to a typical childhood infection while on IL1RA. Alternatively, hepatitis could have represented an atypical presentation of MAS in patients with sJIA taking IL1RA. Further investigation is warranted to determine how anti-IL1 therapies alter immune responsiveness to exogenous triggers in patients with immune dysfunction such as sJIA. Our patients suggest that close monitoring for hepatic and other toxicities is indicated when treating with IL1RA. PMID- 20028521 TI - svmPRAT: SVM-based protein residue annotation toolkit. AB - BACKGROUND: Over the last decade several prediction methods have been developed for determining the structural and functional properties of individual protein residues using sequence and sequence-derived information. Most of these methods are based on support vector machines as they provide accurate and generalizable prediction models. RESULTS: We present a general purpose protein residue annotation toolkit (svmPRAT) to allow biologists to formulate residue-wise prediction problems. svmPRAT formulates the annotation problem as a classification or regression problem using support vector machines. One of the key features of svmPRAT is its ease of use in incorporating any user-provided information in the form of feature matrices. For every residue svmPRAT captures local information around the reside to create fixed length feature vectors. svmPRAT implements accurate and fast kernel functions, and also introduces a flexible window-based encoding scheme that accurately captures signals and pattern for training effective predictive models. CONCLUSIONS: In this work we evaluate svmPRAT on several classification and regression problems including disorder prediction, residue-wise contact order estimation, DNA-binding site prediction, and local structure alphabet prediction. svmPRAT has also been used for the development of state-of-the-art transmembrane helix prediction method called TOPTMH, and secondary structure prediction method called YASSPP. This toolkit developed provides practitioners an efficient and easy-to-use tool for a wide variety of annotation problems. AVAILABILITY: http://www.cs.gmu.edu/~mlbio/svmprat. PMID- 20028522 TI - Behind and beyond the pediatric metabolic syndrome. AB - The growing use of the Metabolic Syndrome in pediatric age need a critical approach, on the basis of recent concerns on definition and usefulness for individual management in clinical practice. We reviewed these aspects from a pediatric point of view, providing a set of questions about what the Metabolic Syndrome means in a clinical setting. The new proposed pediatric definition by IDF was discussed, by outlying how it does not fully consider the peculiarities of children and adolescents. The comparison between two cases of obese children was used in order to show how this diagnosis could be confusing for a correct management. We stressed the need for health-related limits for each component of the Metabolic Syndrome instead of percentile-derived cut-points, as well as the opportunity to extend the estimation to other family or individual risk factors by means of a multiple-items screening form. In conclusion, Metabolic Syndrome use in pediatric age suffers at present from important limitations (i.e., adult derived definition, possibility to rule-in but not to rule-out the individual metabolic risk, instability of MetS during adolescence, poor usefulness of the diagnosis for specific treatment). Consequently, a prudent use of Metabolic Syndrome for children and adolescents seems to be the best and honest position for paediatricians, waiting for long term, longitudinal follow-up studies that could clarify the entire question. PMID- 20028523 TI - Spatiotemporal regulation of the cough motor pattern. AB - The purpose of this study was to identify the spatiotemporal determinants of the cough motor pattern. We speculated that the spatial and temporal characteristics of the cough motor pattern would be regulated separately. Electromyograms (EMG) of abdominal muscles (ABD, rectus abdominis or transversus abdominis), and parasternal muscles (PS) were recorded in anesthetized cats. Repetitive coughing was produced by mechanical stimulation of the lumen of the intrathoracic trachea. Cough inspiratory (CT(I)) and expiratory (CT(E)) durations were obtained from the PS EMG. The ABD EMG burst was confined to the early part of CT(E )and was followed by a quiescent period of varying duration. As such, CT(E )was divided into two segments with CT(E1 )defined as the duration of the ABD EMG burst and CT(E2 )defined as the period of little or no EMG activity in the ABD EMG. Total cough cycle duration (CT(TOT)) was strongly correlated with CT(E2 )(r(2)>0.8), weakly correlated with CT(I )(r(2)<0.3), and not correlated with CT(E1 )(r(2)<0.2). There was no significant relationship between CT(I )and CT(E1 )or CT(E2). The magnitudes of inspiratory and expiratory motor drive during cough were only weakly correlated with each other (r(2)<0.36) and were not correlated with the duration of any phase of cough. The results support: a) separate regulation of CT(I )and CT(E), b) two distinct subphases of CT(E )(CT(E1 )and CT(E2)), c) the duration of CT(E2 )is a primary determinant of CT(TOT), and d) separate regulation of the magnitude and temporal features of the cough motor pattern. PMID- 20028524 TI - Gene cloning and characterization of a novel esterase from activated sludge metagenome. AB - A metagenomic library was prepared using pCC2FOS vector containing about 3.0 Gbp of community DNA from the microbial assemblage of activated sludge. Screening of a part of the un-amplified library resulted in the finding of 1 unique lipolytic clone capable of hydrolyzing tributyrin, in which an esterase gene was identified. This esterase/lipase gene consists of 834 bp and encodes a polypeptide (designated EstAS) of 277 amino acid residuals with a molecular mass of 31 kDa. Sequence analysis indicated that it showed 33% and 31% amino acid identity to esterase/lipase from Gemmata obscuriglobus UQM 2246 (ZP_02733109) and Yarrowia lipolytica CLIB122 (XP_504639), respectively; and several conserved regions were identified, including the putative active site, HSMGG, a catalytic triad (Ser92, His125 and Asp216) and a LHYFRG conserved motif. The EstAS was overexpressed, purified and shown to hydrolyse p-nitrophenyl (NP) esters of fatty acids with short chain lengths (< or = C8). This EstAS had optimal temperature and pH at 35 degrees C and 9.0, respectively, by hydrolysis of p-NP hexanoate. It also exhibited the same level of stability over wide temperature and pH ranges and in the presence of metal ions or detergents. The high level of stability of esterase EstAS with its unique substrate specificities make itself highly useful for biotechnological applications. PMID- 20028525 TI - Biliary sludge and recurrent ketoacidosis: a case report. AB - A five year old boy, weighing 14 kg with no family history of diabetes, presented in frank diabetic ketoacidosis. He recovered, but continued to have episodes of ketoacidosis. He was diagnosed to have biliary sludge, which recovered with insulin treatment. PMID- 20028526 TI - ReseqChip: automated integration of multiple local context probe data from the MitoChip array in mitochondrial DNA sequence assembly. AB - BACKGROUND: The Affymetrix MitoChip v2.0 is an oligonucleotide tiling array for the resequencing of the human mitochondrial (mt) genome. For each of 16,569 nucleotide positions of the mt genome it holds two sets of four 25-mer probes each that match the heavy and the light strand of a reference mt genome and vary only at their central position to interrogate all four possible alleles. In addition, the MitoChip v2.0 carries alternative local context probes to account for known mtDNA variants. These probes have been neglected in most studies due to the lack of software for their automated analysis. RESULTS: We provide ReseqChip, a free software that automates the process of resequencing mtDNA using multiple local context probes on the MitoChip v2.0. ReseqChip significantly improves base call rate and sequence accuracy. ReseqChip is available at http://code.open bio.org/svnweb/index.cgi/bioperl/browse/bioperl-live/trunk/Bio/Microarray/Tools/. CONCLUSIONS: ReseqChip allows for the automated consolidation of base calls from alternative local mt genome context probes. It thereby improves the accuracy of resequencing, while reducing the number of non-called bases. PMID- 20028527 TI - The Medicago truncatula gene expression atlas web server. AB - BACKGROUND: Legumes (Leguminosae or Fabaceae) play a major role in agriculture. Transcriptomics studies in the model legume species, Medicago truncatula, are instrumental in helping to formulate hypotheses about the role of legume genes. With the rapid growth of publically available Affymetrix GeneChip Medicago Genome Array GeneChip data from a great range of tissues, cell types, growth conditions, and stress treatments, the legume research community desires an effective bioinformatics system to aid efforts to interpret the Medicago genome through functional genomics. We developed the Medicago truncatula Gene Expression Atlas (MtGEA) web server for this purpose. DESCRIPTION: The Medicago truncatula Gene Expression Atlas (MtGEA) web server is a centralized platform for analyzing the Medicago transcriptome. Currently, the web server hosts gene expression data from 156 Affymetrix GeneChip(R) Medicago genome arrays in 64 different experiments, covering a broad range of developmental and environmental conditions. The server enables flexible, multifaceted analyses of transcript data and provides a range of additional information about genes, including different types of annotation and links to the genome sequence, which help users formulate hypotheses about gene function. Transcript data can be accessed using Affymetrix probe identification number, DNA sequence, gene name, functional description in natural language, GO and KEGG annotation terms, and InterPro domain number. Transcripts can also be discovered through co-expression or differential expression analysis. Flexible tools to select a subset of experiments and to visualize and compare expression profiles of multiple genes have been implemented. Data can be downloaded, in part or full, in a tabular form compatible with common analytical and visualization software. The web server will be updated on a regular basis to incorporate new gene expression data and genome annotation, and is accessible at: http://bioinfo.noble.org/gene-atlas/. CONCLUSIONS: The MtGEA web server has a well managed rich data set, and offers data retrieval and analysis tools provided in the web platform. It's proven to be a powerful resource for plant biologists to effectively and efficiently identify Medicago transcripts of interest from a multitude of aspects, formulate hypothesis about gene function, and overall interpret the Medicago genome from a systematic point of view. PMID- 20028528 TI - Growth in glucose-based medium and exposure to subinhibitory concentrations of imipenem induce biofilm formation in a multidrug-resistant clinical isolate of Acinetobacter baumannii. AB - BACKGROUND: Acinetobacter baumannii is emerging as an important nosocomial pathogen. Multidrug resistance, as well as ability to withstand environmental stresses, makes eradication of A. baumannii difficult, particularly from hospital settings. RESULTS: Over a six-year period, 73 isolates of A. baumannii were collected from infected patients in two hospitals in Italy. While 69 out of the 73 isolates displayed identical multidrug antibiotic resistance pattern, they were susceptible to carbapenems. Genetic profiles of these 69 isolates, determined by Pulsed Field Gel Electrophoresis (PFGE), indicated that they were genetically related and could be clustered in a specific clone, called SMAL. We tested the ability of the SMAL clone to form biofilm, an important determinant for bacterial colonization of the human host and for persistence in the hospital environment. Biofilm formation by A. baumannii SMAL, measured as surface adhesion to polystyrene, is strongly affected by growth conditions, being impaired in rich growth media such as LB, while being favoured in glucose-based medium. Surface adhesion in glucose-based media is inhibited by treatment with cellulase, suggesting that it depends on production of cellulose or of a chemically related extracellular polysaccharide. Exposure of A. baumannii SMAL to subinhibitory concentrations of imipenem resulted in biofilm stimulation and increased production of iron uptake proteins. Growth in iron-supplemented medium also stimulated surface adhesion, thus suggesting that increased intracellular iron concentrations might act as an environmental signal for biofilm formation in A. baumannii SMAL. CONCLUSIONS: Our results indicate that exposure to subinhibitory concentrations of imipenem can stimulate biofilm formation and induce iron uptake in a pathogenic strain of A. baumannii, with potential implications on antibiotic susceptibility and ability to persist in the human host. PMID- 20028529 TI - Expression of classic cadherins and delta-protocadherins in the developing ferret retina. AB - BACKGROUND: Cadherins are a superfamily of calcium-dependent adhesion molecules that play multiple roles in morphogenesis, including proliferation, migration, differentiation and cell-cell recognition. The subgroups of classic cadherins and delta-protocadherins are involved in processes of neural development, such as neurite outgrowth, pathfinding, target recognition, synaptogenesis as well as synaptic plasticity. We mapped the expression of 7 classic cadherins (CDH4, CDH6, CDH7, CDH8, CDH11, CDH14, CDH20) and 8 delta-protocadherins (PCDH1, PCDH7, PCDH8, PCDH9, PCDH10, PCDH11, PCDH17, PCDH18) at representative stages of retinal development and in the mature retina of the ferret by in situ hybridization. RESULTS: All cadherins investigated by us are expressed differentially by restricted populations of retinal cells during specific periods of the ferret retinogenesis. For example, during embryonic development, some cadherins are exclusively expressed in the outer, proliferative zone of the neuroblast layer, whereas other cadherins mark the prospective ganglion cell layer or cells in the prospective inner nuclear layer. These expression patterns anticipate histogenetic changes that become visible in Nissl or nuclear stainings at later stages. In parallel to the ongoing development of retinal circuits, cadherin expression becomes restricted to specific subpopulations of retinal cell types, especially of ganglion cells, which express most of the investigated cadherins until adulthood. A comparison to previous results in chicken and mouse reveals overall conserved expression patterns of some cadherins but also species differences. CONCLUSIONS: The spatiotemporally restricted expression patterns of 7 classic cadherins and 8 delta-protocadherins indicate that cadherins provide a combinatorial adhesive code that specifies developing retinal cell populations and intraretinal as well as retinofugal neural circuits in the developing ferret retina. PMID- 20028530 TI - Transthoracic echocardiography for imaging of the different coronary artery segments: a feasibility study. AB - BACKGROUND: Transthoracic echocardiography (TTE) may be used for direct inspection of various parts of the main coronary arteries for detection of coronary stenoses and occlusions. We aimed to assess the feasibility of TTE to visualise the complete segments of the left main (LM), left descending (LAD), circumflex (Cx) and right (RCA) coronary arteries. METHODS: One hundred and eleven patients scheduled for diagnostic coronary angiography because of chest pain or acute coronary syndrome had a TTE study to map the passage of the main coronary arteries. LAD, Cx and RCA were each divided into proximal, middle and distal segments. If any part of the individual segment of a coronary artery with antegrade blood flow was not visualised, the segment was labeled as not satisfactorily seen. RESULTS: Complete imaging of the LM was achieved in 98% of the patients. With antegrade directed coronary artery flow, the proximal, middle and distal segments of LAD were completely seen in 96%, 95% and 91% of patients, respectively. Adding the completely seen segments with antegrade coronary flow and segments with retrograde coronary flow, the proximal, middle and distal segments of LAD were adequately visualised in 96%, 96% and 93% of patients, respectively. With antegrade directed coronary artery flow, the proximal, middle and distal segments of Cx were completely seen in 88%, 61% and 3% and in RCA in 40%, 28% and 54% of patients. Retrograde coronary artery flow was correctly identified as verified by coronary angiography in seven coronary segments, mainly in the posterior descending artery (labeled as the distal segment of RCA) and distal LAD. CONCLUSIONS: TTE is a feasible method for complete demonstration of coronary flow in the LM, the proximal Cx and the different segments of LAD, but less suitable for the RCA and mid and distal segments of the Cx. (ClinicalTrials.gov number NTC00281346.). PMID- 20028531 TI - Niche-driven evolution of metabolic and life-history strategies in natural and domesticated populations of Saccharomyces cerevisiae. AB - BACKGROUND: Variation of resource supply is one of the key factors that drive the evolution of life-history strategies, and hence the interactions between individuals. In the yeast Saccharomyces cerevisiae, two life-history strategies related to different resource utilization have been previously described in strains from different industrial origins. In this work, we analyzed metabolic traits and life-history strategies in a broader collection of yeast strains sampled in various ecological niches (forest, human body, fruits, laboratory and industrial environments). RESULTS: By analysing the genetic and plastic variation of six life-history and three metabolic traits, we showed that S. cerevisiae populations harbour different strategies depending on their ecological niches. On one hand, the forest and laboratory strains, referred to as extreme "ants", reproduce quickly, reach a large carrying capacity and a small cell size in fermentation, but have a low reproduction rate in respiration. On the other hand, the industrial strains, referred to as extreme "grasshoppers", reproduce slowly, reach a small carrying capacity but have a big cell size in fermentation and a high reproduction rate in respiration. "Grasshoppers" have usually higher glucose consumption rate than "ants", while they produce lower quantities of ethanol, suggesting that they store cell resources rather than secreting secondary products to cross-feed or poison competitors. The clinical and fruit strains are intermediate between these two groups. CONCLUSIONS: Altogether, these results are consistent with a niche-driven evolution of S. cerevisiae, with phenotypic convergence of populations living in similar habitat. They also revealed that competition between strains having contrasted life-history strategies ("ants" and "grasshoppers") seems to occur at low frequency or be unstable since opposite life-history strategies appeared to be maintained in distinct ecological niches. PMID- 20028532 TI - Soluble iron modulates iron oxide particle-induced inflammatory responses via prostaglandin E(2 )synthesis: In vitro and in vivo studies. AB - BACKGROUND: Ambient particulate matter (PM)-associated metals have been shown to play an important role in cardiopulmonary health outcomes. To study the modulation of PM-induced inflammation by leached off metals, we investigated intracellular solubility of radio-labeled iron oxide ((59)Fe(2)O(3)) particles of 0.5 and 1.5 mum geometric mean diameter. Fe(2)O(3 )particles were examined for the induction of the release of interleukin 6 (IL-6) as pro-inflammatory and prostaglandin E(2 )(PGE(2)) as anti-inflammatory markers in cultured alveolar macrophages (AM) from Wistar Kyoto (WKY) rats. In addition, we exposed male WKY rats to monodispersed Fe(2)O(3 )particles by intratracheal instillation (1.3 or 4.0 mg/kg body weight) to examine in vivo inflammation. RESULTS: Particles of both sizes are insoluble extracellularly in the media but moderately soluble in AM with an intracellular dissolution rate of 0.0037 +/- 0.0014 d(-1 )for 0.5 mum and 0.0016 +/- 0.0012 d(-1 )for 1.5 mum (59)Fe(2)O(3 )particles. AM exposed in vitro to 1.5 mum particles (10 mug/mL) for 24 h increased IL-6 release (1.8-fold; p < 0.05) and also PGE(2 )synthesis (1.9-fold; p < 0.01). By contrast, 0.5 mum particles did not enhance IL-6 release but strongly increased PGE(2 )synthesis (2.5-fold, p < 0.005). Inhibition of PGE(2 )synthesis by indomethacin caused a pro-inflammatory phenotype as noted by increased IL-6 release from AM exposed to 0.5 mum particles (up to 3-fold; p < 0.005). In the rat lungs, 1.5 but not 0.5 mum particles (4.0 mg/kg) induced neutrophil influx and increased vascular permeability. CONCLUSIONS: Fe(2)O(3 )particle-induced neutrophilic inflammatory response in vivo and pro-inflammatory cytokine release in vitro might be modulated by intracellular soluble iron via PGE(2 )synthesis. The suppressive effect of intracellular released soluble iron on particle-induced inflammation has implications on how ambient PM-associated but soluble metals influence pulmonary toxicity of ambient PM. PMID- 20028533 TI - Diagnostic and prognostic value of procalcitonin among febrile critically ill patients with prolonged ICU stay. AB - BACKGROUND: Procalcitonin (PCT) has been proposed as a diagnostic and prognostic sepsis marker, but has never been validated in febrile patients with prolonged ICU stay. METHODS: Patients were included in the study provided they were hospitalised in the ICU for > 10 days, were free of infection and presented a new episode of SIRS, with fever >38 degrees C being obligatory. Fifty patients fulfilled the above criteria. PCT was measured daily during the ICU stay. The primary outcome was proven infection. RESULTS: Twenty-seven out of 50 patients were diagnosed with infection. Median PCT on the day of fever was 1.18 and 0.17 ng/ml for patients with and without proven infections (p < 0.001). The area under the curve for PCT was 0.85 (95% CI; 0.71-0.93), for CRP 0.65 (0.46-0.78) and for WBC 0.68 (0.49-0.81). A PCT level of 1 ng/mL yielded a negative predictive value of 72% for the presence of infection, while a PCT of 1.16 had a specificity of 100%. A two-fold increase of PCT between fever onset and the previous day was associated with proven infection (p 0.001) (OR = 8.55; 2.4-31.1), whereas a four fold increase of PCT of any of the 6 preceding days was associated with a positive predictive value exceeding 69.65%. A PCT value less than 0.5 ng/ml on the third day after the advent of fever was associated with favorable survival (p 0.01). CONCLUSION: The reported data support that serial serum PCT may be a valuable diagnostic and prognostic marker in febrile chronic critically ill patients. PMID- 20028534 TI - Lipids and carotid plaque in the Northern Manhattan Study (NOMAS). AB - BACKGROUND: Lipids, particularly low-density (LDL) and high-density (HDL) lipoproteins, are associated with increased risk of stroke and cardiovascular disease, probably due to atherosclerosis. The objective of this cross-sectional analysis was to investigate the relation between blood lipids and carotid plaque. METHODS: As part of a prospective population-based study to determine the incidence and risk factors of stroke in a multiethnic population, we evaluated 1804 participants with lipid measurements and B-mode ultrasound of carotid arteries (mean age 69 +/- 10 years; 40% men; 51% Hispanic, 26% black, 23% white). The association between lipid parameters and carotid plaque was analyzed by multiple logistic regression. RESULTS: Plaque was present in 61% of participants. Mean total cholesterol was 202 +/- 41 mg/dl. After controlling for other lipid parameters, demographics, and risk factors, the only cholesterol subfraction associated with carotid plaque was LDL (OR per standard deviation (SD) = 1.14, 95% CI 1.02-1.27). Neither HDL nor triglycerides independently predicted carotid plaque. Apolipoprotein B (ApoB) was also associated with risk of plaque (OR per SD = 1.29, 95% CI 1.03-1.60). Apolipoprotein A-I (apoA-1) was associated with a decrease in multiple plaques (OR per SD = 0.76, 95% CI 0.60-0.97), while lipoprotein a was associated with an increased risk of multiple plaques (OR per SD = 1.31, 95% CI 1.03-1.66). ApoB:ApoA-I had the strongest relation with carotid plaque (OR per SD = 1.35, 95% CI 1.08-1.69). CONCLUSIONS: Among the common lipid parameters, LDL has the strongest relation with carotid plaque. Other lipid precursor proteins such as ApoB and ApoA-I may be stronger predictors of subclinical atherosclerosis, however, and better targets for treatment to reduce plaque formation and risk of cerebrovascular disease. PMID- 20028535 TI - Early detection of influenza outbreaks using the DC Department of Health's syndromic surveillance system. AB - BACKGROUND: Since 2001, the District of Columbia Department of Health has been using an emergency room syndromic surveillance system to identify possible disease outbreaks. Data are received from a number of local hospital emergency rooms and analyzed daily using a variety of statistical detection algorithms. The aims of this paper are to characterize the performance of these statistical detection algorithms in rigorous yet practical terms in order to identify the optimal parameters for each and to compare the ability of two syndrome definition criteria and data from a children's hospital versus vs. other hospitals to determine the onset of seasonal influenza. METHODS: We first used a fine-tuning approach to improve the sensitivity of each algorithm to detecting simulated outbreaks and to identifying previously known outbreaks. Subsequently, using the fine-tuned algorithms, we examined (i) the ability of unspecified infection and respiratory syndrome categories to detect the start of the flu season and (ii) how well data from Children's National Medical Center (CNMC) did versus all the other hospitals when using unspecified infection, respiratory, and both categories together. RESULTS: Simulation studies using the data showed that over a range of situations, the multivariate CUSUM algorithm performed more effectively than the other algorithms tested. In addition, the parameters that yielded optimal performance varied for each algorithm, especially with the number of cases in the data stream. In terms of detecting the onset of seasonal influenza, only "unspecified infection," especially the counts from CNMC, clearly delineated influenza outbreaks out of the eight available syndromic classifications. In three of five years, CNMC consistently flags earlier (from 2 days up to 2 weeks earlier) than a multivariate analysis of all other DC hospitals. CONCLUSIONS: When practitioners apply statistical detection algorithms to their own data, fine tuning of parameters is necessary to improve overall sensitivity. With fined tuned algorithms, our results suggest that emergency room based syndromic surveillance focusing on unspecified infection cases in children is an effective way to determine the beginning of the influenza outbreak and could serve as a trigger for more intensive surveillance efforts and initiate infection control measures in the community. PMID- 20028536 TI - Drug procurement, the Global Fund and misguided competition policies. AB - In an effort to increase competition and decrease price, the Global Fund to Fight AIDS, Tuberculosis and Malaria recently began asking some grant recipients to use international competitive bidding processes for certain drug purchases. Unfortunately, for countries like Kenya, this request has caused more harm than good. After awarding the tender for its annual supply of the anti-malarial artemether-lumefantrine to the lowest bidder, Ajanta Pharma, Kenya experienced wide stock-outs in part due to the company's inability to supply the order in full and on time. Similar problems could arise in Uganda. Despite Kenya's experience, Uganda has awarded its next tender for artemether-lumefantrine to Ajanta Pharma. Uganda is already facing wide stock-outs and risks exacerbating an already dire situation the longer it takes to fulfil the procurement contract. A tender process based primarily on price cannot account for a company's ability to consistently supply sufficient product in time. PMID- 20028537 TI - Evidence-informed health policy: are we beginning to get there at last? PMID- 20028538 TI - Evaluation of a novel nanocrystalline hydroxyapatite paste Ostim in comparison to Alpha-BSM - more bone ingrowth inside the implanted material with Ostim compared to Alpha BSM. AB - BACKGROUND: The purpose of this study was to evaluate the performance a newly developed nanocrystalline hydroxyapatite, OSTIM following functional implantation in femoral sites in thirty-eight sheep for 1, 2 or 3 months. Ostim 35 was compared to an established calcium phosphate, Alpha BSM. METHODS: Biomechanical testing, micro-CT analysis, histological and histomorphological analyses were conducted to compare the treatments including evaluation of bone regeneration level, material degradation, implant biomechanical characteristics. RESULTS: The micro-computed tomography (microCT) analysis and macroscopic observations showed that Ostim seemed to diffuse easily particularly when the defects were created in a cancellous bone area. Alpha BSM remained in the defect.The performance of Ostim was good in terms of mechanical properties that were similar to Alpha BSM and the histological analysis showed that the bone regeneration was better with Ostim than with Alpha BSM. The histomorphometric analysis confirmed the qualitative analysis and showed more bone ingrowth inside the implanted material with Ostim when compared to Alpha BSM at all time points. CONCLUSIONS: The successful bone healing with osseous consolidation verifies the importance of the nanocrystalline hydroxyapatite in the treatment of metaphyseal osseous volume defects in the metaphyseal spongiosa. PMID- 20028539 TI - A new measurement of sequence conservation. AB - BACKGROUND: Understanding sequence conservation is important for the study of sequence evolution and for the identification of functional regions of the genome. Current studies often measure sequence conservation based on every position in contiguous regions. Therefore, a large number of functional regions that contain conserved segments separated by relatively long divergent segments are ignored. Our goal in this paper is to define a new measurement of sequence conservation such that both contiguously conserved regions and discontiguously conserved regions can be detected based on this new measurement. Here and in the following, conserved regions are those regions that share similarity higher than a pre-specified similarity threshold with their homologous regions in other species. That is, conserved regions are good candidates of functional regions and may not be always functional. Moreover, conserved regions may contain long and divergent segments. RESULTS: To identify both discontiguously and contiguously conserved regions, we proposed a new measurement of sequence conservation, which measures sequence similarity based only on the conserved segments within the regions. By defining conserved segments using the local alignment tool CHAOS, under the new measurement, we analyzed the conservation of 1642 experimentally verified human functional non-coding regions in the mouse genome. We found that the conservation in at least 11% of these functional regions could be missed by the current conservation analysis methods. We also found that 72% of the mouse homologous regions identified based on the new measurement are more similar to the human functional sequences than the aligned mouse sequences from the UCSC genome browser. We further compared BLAST and discontiguous MegaBLAST with our method. We found that our method picks up many more conserved segments than BLAST and discontiguous MegaBLAST in these regions. CONCLUSIONS: It is critical to have a new measurement of sequence conservation that is based only on the conserved segments in one region. Such a new measurement can aid the identification of better local "orthologous" regions. It will also shed light on the identification of new types of conserved functional regions in vertebrate genomes. PMID- 20028540 TI - A critical assessment of the WHO responsiveness tool: lessons from voluntary HIV testing and counselling services in Kenya. AB - BACKGROUND: Health, fair financing and responsiveness to the user's needs and expectations are seen as the essential objectives of health systems. Efforts have been made to conceptualise and measure responsiveness as a basis for evaluating the non-health aspects of health systems performance. This study assesses the applicability of the responsiveness tool developed by WHO when applied in the context of voluntary HIV counselling and testing services (VCT) at a district level in Kenya. METHODS: A mixed method study was conducted employing a combination of quantitative and qualitative research methods concurrently. The questionnaire proposed by WHO was administered to 328 VCT users and 36 VCT counsellors (health providers). In addition to the questionnaire, qualitative interviews were carried out among a total of 300 participants. Observational field notes were also written. RESULTS: A majority of the health providers and users indicated that the responsiveness elements were very important, e.g. confidentiality and autonomy were regarded by most users and health providers as very important and were also reported as being highly observed in the VCT room. However, the qualitative findings revealed other important aspects related to confidentiality, autonomy and other responsiveness elements that were not captured by the WHO tool. Striking examples were inappropriate location of the VCT centre, limited information provided, language problems, and concern about the quality of counselling. CONCLUSION: The results indicate that the WHO developed responsiveness elements are relevant and important in measuring the performance of voluntary HIV counselling and testing. However, the tool needs substantial revision in order to capture other important dimensions or perspectives. The findings also confirm the importance of careful assessment and recognition of locally specific aspects when conducting comparative studies on responsiveness of HIV testing services. PMID- 20028541 TI - A tagging SNP in INSIG2 is associated with obesity-related phenotypes among Samoans. AB - BACKGROUND: A genome wide association study found significant association of a sequence variant, rs7566605, in the insulin-induced gene 2 (INSIG2) with obesity. However, the association remained inconclusive in follow-up studies. We tested for association of four tagging SNPs (tagSNPs) including this variant with body mass index (BMI) and abdominal circumference (ABDCIR) in the Samoans of the Western Pacific, a population with high levels of obesity. METHODS: We studied 907 adult Samoan participants from a longitudinal study of adiposity and cardiovascular disease risk in two polities, American Samoa and Samoa. Four tagSNPs were identified from the Chinese HapMap database based on pairwise r2 of >or=0.8 and minor allele frequency of >or=0.05. Genotyping was performed using the TaqMan assay. Tests of association with BMI and ABDCIR were performed under the additive model. RESULTS: We did not find association of rs7566605 with either BMI or ABDCIR in any group of the Samoans. However, the most distally located tagSNPs in Intron 3 of the gene, rs9308762, showed significant association with both BMI (p-value 0.024) and ABDCIR (p-value 0.009) in the combined sample and with BMI (p-value 0.038) in the sample from Samoa. CONCLUSION: Although rs7566605 was not significantly associated with obesity in our study population, we can not rule out the involvement of INSIG2 in obesity related traits as we found significant association of another tagSNP in INSIG2 with both BMI and ABDCIR. This study suggests the importance of comprehensive assessment of sequence variants within a gene in association studies. PMID- 20028542 TI - A blind deconvolution approach to high-resolution mapping of transcription factor binding sites from ChIP-seq data. AB - We present CSDeconv, a computational method that determines locations of transcription factor binding from ChIP-seq data. CSDeconv differs from prior methods in that it uses a blind deconvolution approach that allows closely-spaced binding sites to be called accurately. We apply CSDeconv to novel ChIP-seq data for DosR binding in Mycobacterium tuberculosis and to existing data for GABP in humans and show that it can discriminate binding sites separated by as few as 40 bp. PMID- 20028543 TI - Comparative attainment of 5-year undergraduate and 4-year graduate entry medical students moving into foundation training. AB - BACKGROUND: Graduate entry medicine is a recent innovation in UK medical training. Evidence is sparse at present as to progress and attainment on these programmes. Shared clinical rotations, between an established 5-year and a new graduate entry course, provide the opportunity to compare achievement on clinical assessments. To compare completion and attainment on clinical phase assessments between students on a 4-year graduate entry course and an established 5-year undergraduate medicine course. METHODS: Overall completion rates for the 4 and 5 year courses, fails at first attempt, and scores on 14 clinical assessments, were compared between 171 graduate-entry and 450 undergraduate medical students at the University of Nottingham, comprising two graduating cohorts. Percentage assessment marks were converted to z-scores separately for each graduating year and the normalised marks then combined into a single dataset. Z-score transformed percentage marks were analysed by multivariate analysis of variance and univariate analyses of variance for each summative assessment. Numbers of fails at first attempt were analysed aggregated across all assessments initially, then separately for each assessment using chi2. RESULTS: Completion rates were around 90% overall and significantly higher in the graduate entry course. Failures of assessments overall were similar, but a higher proportion of graduate entry students failed the final OSLER. Mean performance on clinical assessments showed a significant overall difference, made up of lower performance on 4 of 5 knowledge-based exams (as well as higher performance on the first exam) by the graduate entry group, but similar levels of performance on all the skills-based and attitudinal assessments. CONCLUSIONS: High completion rates are encouraging. The lower performance in some knowledge-based exams may reflect lower prior educational attainment, a substantially different demographic profile (age, gender), or an artefact of the first 2 years of a new graduate entry programme. PMID- 20028544 TI - Formulation preference, tolerability and quality of life assessment following a switch from lopinavir/ritonavir soft gel capsule to tablet in human immunodeficiency virus-infected patients. AB - BACKGROUND: Lopinavir/ritonavir (LPV/r) tablet compared to the soft gel capsule (SGC) formulation has no oleic acid or sorbitol, has no refrigeration or food restriction requirements, and has less pharmacokinetic variability. We compared the tolerability, quality of life (QoL), and formulation preference after switching from LPV/r SGC to the tablet formulation. METHODS: In a prospective, single-arm, cohort study-design, 74 human immunodeficiency virus (HIV) infected subjects stable on LPV/r-based therapy were enrolled prior to (n = 25) or 8 weeks (n = 49) after switching from SGC to tablet. Baseline data included clinical laboratory tests, bowel habit survey (BHS) and QoL questionnaire (recalled if enrolled post-switch). Global Condition Improvement (GCI)-score, BHS-score, QoL score, and formulation preference data were captured at weeks 4 and 12. RESULTS: At week 12 post-enrollment; the tablet was preferred to the SGC (74% vs. 10%, p < 0.0001). GCI-overall-tolerability score was 2.46 +/- 3.30 on a scale of -7 to +7, with 90% admitting to feeling better or about the same. Stool frequency, consistency, volume, and +/- blood improved, however the improvement was significant in "consistency" only (p = 0.03). Aggregate Bowel Habit-Profile improved (BHS-score change = -0.227, p = 0.01). Inverse relationship existed between GCI and BHS (slope = -1.2, p = 0.02) at week-4, suggesting that improved overall-tolerability was related to better gastrointestinal (GI)-tolerance. QoL scores were stable. Mean reductions in total cholesterol of 9.20 mg/dL (p = 0.02), in triglycerides of 33 mg/dL (p = 0.04), and in HDL of 4.50 mg/dL (p = 0.01) unrelated to lipid-lowering therapy, were observed at week 12. CONCLUSIONS: LPV/r-tablet was well tolerated and preferred to the SGC in HIV infected subjects, with stable QoL and appreciable improvement in GI-tolerability. The unexpected changes in lipid profile deserve further evaluation. PMID- 20028545 TI - Characterization of some Brucella species from Zimbabwe by biochemical profiling and AMOS-PCR. AB - BACKGROUND: Bovine brucellosis caused by Brucella abortus is endemic in most large commercial and smallholder cattle farms of Zimbabwe, while brucellosis in other domestic animals is rare. The diagnosis of brucellosis is mainly accomplished using serological tests. However, some Brucella spp. have been isolated from clinical cases in the field and kept in culture collection but their biochemical profiles were not documented. We report biochemical profiling and AMOS-PCR characterization of some of these field isolates of Brucella originating from both commercial and smallholder cattle farming sectors of Zimbabwe. FINDINGS: Fourteen isolates of Brucella from culture collection were typed using biochemical profiles, agglutination by monospecific antisera, susceptibility to Brucella-specific bacteriophages and by AMOS-PCR that amplifies species- specific IS711. The results of the biochemical profiles for B. abortus biovar 1 (11 isolates) and biovar 2 (2 isolates) were consistent with those of reference strains. A single isolate from a goat originating from a smallholder mixed animal farm was identified as B. melitensis biovar 1. The AMOS-PCR produced DNA products of sizes 498 bp and 731 bp for B. abortus (biovar 1 and 2) and B. melitensis biovar 1, respectively. CONCLUSION: We concluded that the biochemical profiles and AMOS-PCR characterization were consistent with their respective species and biovars. B. abortus biovar 1 is likely to be the predominant cause of brucellosis in both commercial and smallholder cattle farms in Zimbabwe. PMID- 20028546 TI - From uncertainty to reward: BOLD characteristics differentiate signaling pathways. AB - BACKGROUND: Reward value and uncertainty are represented by dopamine neurons in monkeys by distinct phasic and tonic firing rates. Knowledge about the underlying differential dopaminergic pathways is crucial for a better understanding of dopamine-related processes. Using functional magnetic resonance blood-oxygen level dependent (BOLD) imaging we analyzed brain activation in 15 healthy, male subjects performing a gambling task, upon expectation of potential monetary rewards at different reward values and levels of uncertainty. RESULTS: Consistent with previous studies, ventral striatal activation was related to both reward magnitudes and values. Activation in medial and lateral orbitofrontal brain areas was best predicted by reward uncertainty. Moreover, late BOLD responses relative to trial onset were due to expectation of different reward values and likely to represent phasic dopaminergic signaling. Early BOLD responses were due to different levels of reward uncertainty and likely to represent tonic dopaminergic signals. CONCLUSIONS: We conclude that differential dopaminergic signaling as revealed in animal studies is not only represented locally by involvement of distinct brain regions but also by distinct BOLD signal characteristics. PMID- 20028547 TI - Progressive colonization and restricted gene flow shape island-dependent population structure in Galapagos marine iguanas (Amblyrhynchus cristatus). AB - BACKGROUND: Marine iguanas (Amblyrhynchus cristatus) inhabit the coastlines of large and small islands throughout the Galapagos archipelago, providing a rich system to study the spatial and temporal factors influencing the phylogeographic distribution and population structure of a species. Here, we analyze the microevolution of marine iguanas using the complete mitochondrial control region (CR) as well as 13 microsatellite loci representing more than 1200 individuals from 13 islands. RESULTS: CR data show that marine iguanas occupy three general clades: one that is widely distributed across the northern archipelago, and likely spread from east to west by way of the South Equatorial current, a second that is found mostly on the older eastern and central islands, and a third that is limited to the younger northern and western islands. Generally, the CR haplotype distribution pattern supports the colonization of the archipelago from the older, eastern islands to the younger, western islands. However, there are also signatures of recurrent, historical gene flow between islands after population establishment. Bayesian cluster analysis of microsatellite genotypes indicates the existence of twenty distinct genetic clusters generally following a one-cluster-per-island pattern. However, two well-differentiated clusters were found on the easternmost island of San Cristobal, while nine distinct and highly intermixed clusters were found on youngest, westernmost islands of Isabela and Fernandina. High mtDNA and microsatellite genetic diversity were observed for populations on Isabela and Fernandina that may be the result of a recent population expansion and founder events from multiple sources. CONCLUSIONS: While a past genetic study based on pure FST analysis suggested that marine iguana populations display high levels of nuclear (but not mitochondrial) gene flow due to male-biased dispersal, the results of our sex-biased dispersal tests and the finding of strong genetic differentiation between islands do not support this view. Therefore, our study is a nice example of how recently developed analytical tools such as Bayesian clustering analysis and DNA sequence-based demographic analyses can overcome potential biases introduced by simply relying on FST estimates from markers with different inheritance patterns. PMID- 20028548 TI - Antigen-specific influence of GM/KM allotypes on IgG isotypes and association of GM allotypes with susceptibility to Plasmodium falciparum malaria. AB - BACKGROUND: Plasmodium falciparum malaria is a complex disease in which genetic and environmental factors influence susceptibility. IgG isotypes are in part genetically controlled, and GM/KM allotypes are believed to be involved in this control. METHODS: In this study, 216 individuals from Daraweesh, an area of seasonal malaria transmission in Sudan, were followed for nine years for malaria infection. Total IgG and IgG isotypes against four malaria antigens, MSP2-3D7, MSP2-FC27, AMA1, and Pf332-C231 were measured in plasma obtained from the cohort at the end of the study, during the dry malaria-free period. The GM/KM allotypes of the donors were determined. RESULTS: The GM 1,17 5,13,14,6 phenotype was associated with a higher incidence of malaria compared with the non-1,17 5,13,14,6 phenotypes (P = 0.037). Paradoxically, the carriers of the GM 1,17 5,13,14,6 phenotype had significantly higher baseline levels of total IgG and non cytophilic IgG isotypes as compared to non-carriers. The KM allotypes influence on IgG isotypes level was limited. Finally, the differences in the baseline concentrations of total IgG and IgG isotypes between the different GK/KM phenotype carriers were antigen-dependent. DISCUSSION: The results show that GM but not KM allotypes appeared to influence host susceptibility to uncomplicated malaria as well as the antibody profile of the donors, and the carriers of the GM 1,17 5,13,14,6 phenotype were the most susceptible CONCLUSIONS: The GM allotypes have significant influence on susceptibility to uncomplicated P. falciparum malaria and antigen-dependent influence on total IgG and IgG subclasses. PMID- 20028549 TI - Phylogenetic analysis and temporal diversification of mosquitoes (Diptera: Culicidae) based on nuclear genes and morphology. AB - BACKGROUND: Phylogenetic analyses provide a framework for examining the evolution of morphological and molecular diversity, interpreting patterns in biogeography, and achieving a stable classification. The generic and suprageneric relationships within mosquitoes (Diptera: Culicidae) are poorly resolved, making these subjects difficult to address. RESULTS: We carried out maximum parsimony and maximum likelihood, including Bayesian, analyses on a data set consisting of six nuclear genes and 80 morphological characters to assess their ability to resolve relationships among 25 genera. We also estimated divergence times based on sequence data and fossil calibration points, using Bayesian relaxed clock methods. Strong support was recovered for the basal position and monophyly of the subfamily Anophelinae and the tribes Aedini and Sabethini of subfamily Culicinae. Divergence times for major culicid lineages date to the early Cretaceous. CONCLUSIONS: Deeper relationships within the family remain poorly resolved, suggesting the need for additional taxonomic sampling. Our results support the notion of rapid radiations early in the diversification of mosquitoes. PMID- 20028550 TI - Cellular adaptive immune response against porcine circovirus type 2 in subclinically infected pigs. AB - BACKGROUND: Porcine circovirus type 2 (PCV2) is a dominant causative agent of postweaning multisystemic wasting syndrome (PMWS), a multifactorial disease complex with putative immunosuppressive characteristics. Little is known about adaptive PCV2-specific immune responses in infected pigs. Therefore, the T and B cell responses following PCV2 infection in 3-week old SPF piglets infected with PCV2 or PCV2 plus porcine parvovirus (PPV) were studied. RESULTS: All animals were asymptomatically infected. At 7 days post infection (d p.i.), B lymphocyte and T lymphocyte numbers decreased in the dual infected, but not in the single infected piglets. At this time point a transient PCV2 viraemia was noted in the PCV2 infected groups. Antibodies against the infecting virus were detectable at day 24-28 p.i. for anti-PCV2 antibodies and at day 10 p.i. for anti-PPV antibodies, with no apparent influence of PCV2 on the early PPV antibody development. In the animals infected with PPV alone, IFN-gamma secreting cells (SC) that were not specific for PCV2 were detected by ELISPOT assay at day 7 p.i. Interestingly, this response was absent in the PCV2/PPV dual infected animals. PCV2-specific IFN-gamma SC were observed in the PCV2/PPV infected group at 7 d p.i. and in the PCV2 single infected group at 21 d p.i. A reduction in the numbers of IFN-gamma SC was observed following anti-CD4 and anti-CD8 antibody treatment, suggesting roles for both CD4+ and CD8+ T cells in the response against PCV2 infection. This was supported by an observed increase in the percentage of IFN-gamma positive CD8hi cytotoxic T cells as well as IFN-gamma positive CD8-/low helper T cells after PCV2 in vitro re-stimulation. CONCLUSIONS: Infection of weaned SPF piglets with PCV2 alone or combined with PPV does not induce disease and in both cases a relatively slow anti-PCV2 antibody response and weak T lymphocyte responses were found. Knowledge on such immunological characteristics is important for both PCV2 pathogenesis and vaccination. PMID- 20028551 TI - Association of the components of the metabolic syndrome with non-alcoholic fatty liver disease among normal-weight, overweight and obese children and adolescents. AB - OBJECTIVES: This study aimed to determine the prevalence of the metabolic syndrome, abnormalities of liver enzymes and sonographic fatty liver, as well as the inter-related associations in normal weight, overweight and obese children and adolescents. METHODS: This cross-sectional study was conducted among a sample of 1107 students (56.1% girls), aged 6-18 years in Isfahan, Iran. In addition to physical examination, fasting blood glucose, serum lipid profile and liver enzymes were determined. Liver sonography was performed among 931 participants. These variables were compared among participants with different body mass index (BMI) categories. RESULTS: From lower to higher BMI category, alanine aminotransferase (ALT), total cholesterol, LDL-cholesterol, triglycerides and systolic blood pressure increased, and HDL-cholesterol decreased significantly. Elevated ALT, aspartate aminotransferase (AST) and alkaline phosphatase (ALP) were documented in respectively 4.1%, 6.6% and 9.8% of normal weight group. The corresponding figure was 9.5%, 9.8% and 9.1% in overweight group, and 16.9%, 14.9% and 10.8% in obese group, respectively. In all BMI categories, ALT increased significantly by increasing the number of the components of the metabolic syndrome. Odds ratio for elevated liver enzymes and sonographic fatty liver increased significantly with higher number of the components of the metabolic syndrome and higher BMI categories before and after adjustment for age. CONCLUSIONS: Because of the interrelationship of biochemical and sonographic indexes of fatty liver with the components of the metabolic syndrome, and with increase in their number, it is suggested to determine the clinical impact of such association in future longitudinal studies. PMID- 20028552 TI - Computational modeling of the EGFR network elucidates control mechanisms regulating signal dynamics. AB - BACKGROUND: The epidermal growth factor receptor (EGFR) signaling pathway plays a key role in regulation of cellular growth and development. While highly studied, it is still not fully understood how the signal is orchestrated. One of the reasons for the complexity of this pathway is the extensive network of inter connected components involved in the signaling. In the aim of identifying critical mechanisms controlling signal transduction we have performed extensive analysis of an executable model of the EGFR pathway using the stochastic pi calculus as a modeling language. RESULTS: Our analysis, done through simulation of various perturbations, suggests that the EGFR pathway contains regions of functional redundancy in the upstream parts; in the event of low EGF stimulus or partial system failure, this redundancy helps to maintain functional robustness. Downstream parts, like the parts controlling Ras and ERK, have fewer redundancies, and more than 50% inhibition of specific reactions in those parts greatly attenuates signal response. In addition, we suggest an abstract model that captures the main control mechanisms in the pathway. Simulation of this abstract model suggests that without redundancies in the upstream modules, signal transduction through the entire pathway could be attenuated. In terms of specific control mechanisms, we have identified positive feedback loops whose role is to prolong the active state of key components (e.g., MEK-PP, Ras-GTP), and negative feedback loops that help promote signal adaptation and stabilization. CONCLUSIONS: The insights gained from simulating this executable model facilitate the formulation of specific hypotheses regarding the control mechanisms of the EGFR signaling, and further substantiate the benefit to construct abstract executable models of large complex biological networks. PMID- 20028553 TI - The journey of Diagnostic Pathology in 2009, perspectives for 2010. PMID- 20028554 TI - G+C content dominates intrinsic nucleosome occupancy. AB - BACKGROUND: The relative preference of nucleosomes to form on individual DNA sequences plays a major role in genome packaging. A wide variety of DNA sequence features are believed to influence nucleosome formation, including periodic dinucleotide signals, poly-A stretches and other short motifs, and sequence properties that influence DNA structure, including base content. It was recently shown by Kaplan et al. that a probabilistic model using composition of all 5-mers within a nucleosome-sized tiling window accurately predicts intrinsic nucleosome occupancy across an entire genome in vitro. However, the model is complicated, and it is not clear which specific DNA sequence properties are most important for intrinsic nucleosome-forming preferences. RESULTS: We find that a simple linear combination of only 14 simple DNA sequence attributes (G+C content, two transformations of dinucleotide composition, and the frequency of eleven 4-bp sequences) explains nucleosome occupancy in vitro and in vivo in a manner comparable to the Kaplan model. G+C content and frequency of AAAA are the most important features. G+C content is dominant, alone explaining approximately 50% of the variation in nucleosome occupancy in vitro. CONCLUSIONS: Our findings provide a dramatically simplified means to predict and understand intrinsic nucleosome occupancy. G+C content may dominate because it both reduces frequency of poly-A-like stretches and correlates with many other DNA structural characteristics. Since G+C content is enriched or depleted at many types of features in diverse eukaryotic genomes, our results suggest that variation in nucleotide composition may have a widespread and direct influence on chromatin structure. PMID- 20028555 TI - Potential impact of stress activated retrotransposons on genome evolution in a marine diatom. AB - BACKGROUND: Transposable elements (TEs) are mobile DNA sequences present in the genomes of most organisms. They have been extensively studied in animals, fungi, and plants, and have been shown to have important functions in genome dynamics and species evolution. Recent genomic data can now enlarge the identification and study of TEs to other branches of the eukaryotic tree of life. Diatoms, which belong to the heterokont group, are unicellular eukaryotic algae responsible for around 40% of marine primary productivity. The genomes of a centric diatom, Thalassiosira pseudonana, and a pennate diatom, Phaeodactylum tricornutum, that likely diverged around 90 Mya, have recently become available. RESULTS: In the present work, we establish that LTR retrotransposons (LTR-RTs) are the most abundant TEs inhabiting these genomes, with a much higher presence in the P. tricornutum genome. We show that the LTR-RTs found in diatoms form two new phylogenetic lineages that appear to be diatom specific and are also found in environmental samples taken from different oceans. Comparative expression analysis in P. tricornutum cells cultured under 16 different conditions demonstrate high levels of transcriptional activity of LTR retrotransposons in response to nitrate limitation and upon exposure to diatom-derived reactive aldehydes, which are known to induce stress responses and cell death. Regulatory aspects of P. tricornutum retrotransposon transcription also include the occurrence of nitrate limitation sensitive cis-regulatory components within LTR elements and cytosine methylation dynamics. Differential insertion patterns in different P. tricornutum accessions isolated from around the world infer the role of LTR-RTs in generating intraspecific genetic variability. CONCLUSION: Based on these findings we propose that LTR-RTs may have been important for promoting genome rearrangements in diatoms. PMID- 20028556 TI - Scale-up from microtiter plate to laboratory fermenter: evaluation by online monitoring techniques of growth and protein expression in Escherichia coli and Hansenula polymorpha fermentations. AB - BACKGROUND: In the past decade, an enormous number of new bioprocesses have evolved in the biotechnology industry. These bioprocesses have to be developed fast and at a maximum productivity. Up to now, only few microbioreactors were developed to fulfill these demands and to facilitate sample processing. One predominant reaction platform is the shaken microtiter plate (MTP), which provides high-throughput at minimal expenses in time, money and work effort. By taking advantage of this simple and efficient microbioreactor array, a new online monitoring technique for biomass and fluorescence, called BioLector, has been recently developed. The combination of high-throughput and high information content makes the BioLector a very powerful tool in bioprocess development. Nevertheless, the scalabilty of results from the micro-scale to laboratory or even larger scales is very important for short development times. Therefore, engineering parameters regarding the reactor design and its operation conditions play an important role even on a micro-scale. In order to evaluate the scale-up from a microtiter plate scale (200 microL) to a stirred tank fermenter scale (1.4 L), two standard microbial expression systems, Escherichia coli and Hansenula polymorpha, were fermented in parallel at both scales and compared with regard to the biomass and protein formation. RESULTS: Volumetric mass transfer coefficients (kLa) ranging from 100 to 350 1/h were obtained in 96-well microtiter plates. Even with a suboptimal mass transfer condition in the microtiter plate compared to the stirred tank fermenter (kLa = 370-600 1/h), identical growth and protein expression kinetics were attained in bacteria and yeast fermentations. The bioprocess kinetics were evaluated by optical online measurements of biomass and protein concentrations exhibiting the same fermentation times and maximum signal deviations below 10% between the scales. In the experiments, the widely applied green fluorescent protein (GFP) served as an online reporter of protein expression for both strains. CONCLUSIONS: The successful 7000-fold scale-up from a shaken microtiter plate to a stirred tank fermenter was demonstrated in parallel fermentations for standard microbial expression systems. This confirms that the very economical and time efficient platform of microtiter plates can be very easily scaled up to larger stirred tank fermenters under defined engineering conditions. New online monitoring techniques for microtiter plates, such as the BioLector, provide even more real-time kinetic data from fermentations than ever before and at an affordable price. This paves the way for a better understanding of the bioprocess and a more rational process design. PMID- 20028557 TI - The c-Met receptor tyrosine kinase inhibitor MP470 radiosensitizes glioblastoma cells. AB - PURPOSE: Glioblastoma multiforme (GBM) is resistant to current cytotoxic therapies, in part because of enhanced DNA repair. Activation of the receptor tyrosine kinase c-Met has been shown to protect cancer cells from DNA damage. We hypothesized that inhibiting c-Met would decrease this protection and thus sensitize resistant tumor cells to the effects of radiation therapy. MATERIALS AND METHODS: Eight human GBM cell lines were screened for radiosensitivity to the small-molecule c-Met inhibitor MP470 with colony-count assays. Double-strand (ds) DNA breaks was quantified by using antibodies to gamma H2AX. Western blotting demonstrate expression of RAD51, glycogen synthase kinase (GSK)-3beta, and other proteins. A murine xenograft tumor flank model was used for in vivo radiosensitization studies. RESULTS: MP470 reduced c-Met phosphorylation and enhanced radiation-induced cell kill by 0.4 logs in SF767 cells. Cells pretreated with MP470 had more ds DNA damage than cells treated with radiation alone. Mechanistically, MP470 was shown to inhibit dsDNA break repair and increase apoptosis. MP470 influences various survival and DNA repair related proteins such as pAKT, RAD51 and GSK3beta. In vivo, the addition of MP470 to radiation resulted in a tumor-growth-delay enhancement ratio of 2.9 over radiation alone and extended survival time. CONCLUSIONS: GBM is a disease site where radiation is often used to address both macroscopic and microscopic disease. Despite attempts at dose escalation outcomes remain poor. MP470, a potent small-molecule tyrosine kinase inhibitor of c-Met, radiosensitized several GBM cell lines both in vitro and in vivo, and may help to improve outcomes for patients with GBM. PMID- 20028558 TI - Canine tumor cross-species genomics uncovers targets linked to osteosarcoma progression. AB - BACKGROUND: Pulmonary metastasis continues to be the most common cause of death in osteosarcoma. Indeed, the 5-year survival for newly diagnosed osteosarcoma patients has not significantly changed in over 20 years. Further understanding of the mechanisms of metastasis and resistance for this aggressive pediatric cancer is necessary. Pet dogs naturally develop osteosarcoma providing a novel opportunity to model metastasis development and progression. Given the accelerated biology of canine osteosarcoma, we hypothesized that a direct comparison of canine and pediatric osteosarcoma expression profiles may help identify novel metastasis-associated tumor targets that have been missed through the study of the human cancer alone. RESULTS: Using parallel oligonucleotide array platforms, shared orthologues between species were identified and normalized. The osteosarcoma expression signatures could not distinguish the canine and human diseases by hierarchical clustering. Cross-species target mining identified two genes, interleukin-8 (IL-8) and solute carrier family 1 (glial high affinity glutamate transporter), member 3 (SLC1A3), which were uniformly expressed in dog but not in all pediatric osteosarcoma patient samples. Expression of these genes in an independent population of pediatric osteosarcoma patients was associated with poor outcome (p = 0.020 and p = 0.026, respectively). Validation of IL-8 and SLC1A3 protein expression in pediatric osteosarcoma tissues further supported the potential value of these novel targets. Ongoing evaluation will validate the biological significance of these targets and their associated pathways. CONCLUSIONS: Collectively, these data support the strong similarities between human and canine osteosarcoma and underline the opportunities provided by a comparative oncology approach as a means to improve our understanding of cancer biology and therapies. PMID- 20028559 TI - Habitat suitability and ecological niche profile of major malaria vectors in Cameroon. AB - BACKGROUND: Suitability of environmental conditions determines a species distribution in space and time. Understanding and modelling the ecological niche of mosquito disease vectors can, therefore, be a powerful predictor of the risk of exposure to the pathogens they transmit. In Africa, five anophelines are responsible for over 95% of total malaria transmission. However, detailed knowledge of the geographic distribution and ecological requirements of these species is to date still inadequate. METHODS: Indoor-resting mosquitoes were sampled from 386 villages covering the full range of ecological settings available in Cameroon, Central Africa. Using a predictive species distribution modeling approach based only on presence records, habitat suitability maps were constructed for the five major malaria vectors Anopheles gambiae, Anopheles funestus, Anopheles arabiensis, Anopheles nili and Anopheles moucheti. The influence of 17 climatic, topographic, and land use variables on mosquito geographic distribution was assessed by multivariate regression and ordination techniques. RESULTS: Twenty-four anopheline species were collected, of which 17 are known to transmit malaria in Africa. Ecological Niche Factor Analysis, Habitat Suitability modeling and Canonical Correspondence Analysis revealed marked differences among the five major malaria vector species, both in terms of ecological requirements and niche breadth. Eco-geographical variables (EGVs) related to human activity had the highest impact on habitat suitability for the five major malaria vectors, with areas of low population density being of marginal or unsuitable habitat quality. Sunlight exposure, rainfall, evapo transpiration, relative humidity, and wind speed were among the most discriminative EGVs separating "forest" from "savanna" species. CONCLUSIONS: The distribution of major malaria vectors in Cameroon is strongly affected by the impact of humans on the environment, with variables related to proximity to human settings being among the best predictors of habitat suitability. The ecologically more tolerant species An. gambiae and An. funestus were recorded in a wide range of eco-climatic settings. The other three major vectors, An. arabiensis, An. moucheti, and An. nili, were more specialized. Ecological niche and species distribution modelling should help improve malaria vector control interventions by targeting places and times where the impact on vector populations and disease transmission can be optimized. PMID- 20028560 TI - Generation and analysis of expression sequence tags from haustoria of the wheat stripe rust fungus Puccinia striiformis f. sp. Tritici. AB - BACKGROUND: Stripe rust, caused by Puccinia striiformis f. sp. tritici (Pst), is one of the most destructive diseases of wheat (Triticum aestivum L.) worldwide. In spite of its agricultural importance, the genomics and genetics of the pathogen are poorly characterized. Pst transcripts from urediniospores and germinated urediniospores have been examined previously, but little is known about genes expressed during host infection. Some genes involved in virulence in other rust fungi have been found to be specifically expressed in haustoria. Therefore, the objective of this study was to generate a cDNA library to characterize genes expressed in haustoria of Pst. RESULTS: A total of 5,126 EST sequences of high quality were generated from haustoria of Pst, from which 287 contigs and 847 singletons were derived. Approximately 10% and 26% of the 1,134 unique sequences were homologous to proteins with known functions and hypothetical proteins, respectively. The remaining 64% of the unique sequences had no significant similarities in GenBank. Fifteen genes were predicted to be proteins secreted from Pst haustoria. Analysis of ten genes, including six secreted protein genes, using quantitative RT-PCR revealed changes in transcript levels in different developmental and infection stages of the pathogen. CONCLUSIONS: The haustorial cDNA library was useful in identifying genes of the stripe rust fungus expressed during the infection process. From the library, we identified 15 genes encoding putative secreted proteins and six genes induced during the infection process. These genes are candidates for further studies to determine their functions in wheat-Pst interactions. PMID- 20028561 TI - microRNA evaluation of unknown primary lesions in the head and neck. AB - Unknown primary malignancy in the head and neck is not an infrequent diagnosis for patients with metastatic cervical lymph nodes. Although linked with a relatively good prognosis following radiation treatment, widespread radiation is coupled with significant morbidity. Altered microRNA (miRNA) expression has been associated with both cancer progression and metastasis. We sought to determine whether miRNA expression analysis could be used as a diagnostic tool to discover the primary site of malignancy, within the head and neck. We used quantitative real-time PCR to identify miRNA expression profiles of squamous cell carcinoma of the tonsil, base of tongue and post-nasal space, as well as their corresponding metastatic lymph nodes, from 6 patients. Our results revealed that each cancer maintained its expression profile between the primary site and the nodal metastasis (r = 0.82, p < 0.0001). In addition, each anatomical sub-site maintained a distinct miRNA profile between individual patients (r = 0.79, p < 0.0001). Finally, between sub-sites, the miRNA profiles were distinct (p < 0.0001). As proof of principle, our study provides an indication that miRNA expression analysis may be useful to compare the primary lesion and local metastatic disease. This may be clinically relevant to predict the primary site of origin of metastatic disease, when the primary site remains obscure. PMID- 20028562 TI - Gain in cellular organization of inflammatory breast cancer: A 3D in vitro model that mimics the in vivo metastasis. AB - BACKGROUND: The initial step of metastasis in carcinomas, often referred to as the epithelial-mesenchymal transition (EMT), occurs via the loss of adherens junctions (e.g. cadherins) by the tumor embolus. This leads to a subsequent loss of cell polarity and cellular differentiation and organization, enabling cells of the embolus to become motile and invasive. However highly malignant inflammatory breast cancer (IBC) over-expresses E-cadherin. The human xenograft model of IBC (MARY-X), like IBC, displays the signature phenotype of an exaggerated degree of lymphovascular invasion (LVI) in situ by tumor emboli. An intact E cadherin/alpha, beta-catenin axis mediates the tight, compact clump of cells found both in vitro and in vivo as spheroids and tumor emboli, respectively. METHODS: Using electron microscopy and focused ion beam milling to acquire in situ sections, we performed ultrastructural analysis of both an IBC and non-IBC, E-cadherin positive cell line to determine if retention of this adhesion molecule contributed to cellular organization. RESULTS: Here we report through ultrastructural analysis that IBC exhibits a high degree of cellular organization with polar elements such as apical/lateral positioning of E-cadherin, apical surface microvilli, and tortuous lumen-like (canalis) structures. In contrast, agarose-induced spheroids of MCF-7, a weakly invasive E-cadherin positive breast carcinoma cell line, do not exhibit ultrastructural polar features. CONCLUSIONS: This study has determined that the highly metastatic IBC with an exaggerated malignant phenotype challenges conventional wisdom in that instead of displaying a loss of cellular organization, IBC acquires a highly structured architecture.These findings suggest that the metastatic efficiency might be linked to the formation and maintenance of these architectural features. The comparative architectural features of both the spheroid and embolus of MARY-X provide an in vitro model with tractable in vivo applications. PMID- 20028563 TI - Use and limitations of malaria rapid diagnostic testing by community health workers in war-torn Democratic Republic of Congo. AB - BACKGROUND: Accurate and practical malaria diagnostics, such as immunochromatographic rapid diagnostic tests (RDTs), have the potential to avert unnecessary treatments and save lives. Volunteer community health workers (CHWs) represent a potentially valuable human resource for expanding this technology to where it is most needed, remote rural communities in sub-Saharan Africa with limited health facilities and personnel. This study reports on a training programme for CHWs to incorporate RDTs into their management strategy for febrile children in the Democratic Republic of Congo, a tropical African setting ravaged by human conflict. METHODS: Prospective cohort study, satisfaction questionnaire and decision analysis. RESULTS: Twelve CHWs were trained to safely and accurately perform and interpret RDTs, then successfully implemented rapid diagnostic testing in their remote community in a cohort of 357 febrile children. CHWs were uniformly positive in evaluating RDTs for their utility and ease of use. However, high malaria prevalence in this cohort (93% by RDTs, 88% by light microscopy) limited the cost-effectiveness of RDTs compared to presumptive treatment of all febrile children, as evidenced by findings from a simplified decision analysis. CONCLUSIONS: CHWs can safely and effectively use RDTs in their management of febrile children; however, cost-effectiveness of RDTs is limited in zones of high malaria prevalence. PMID- 20028564 TI - Geochemical characterization of oceanic basalts using Artificial Neural Network. AB - The geochemical discriminate diagrams help to distinguish the volcanics recovered from different tectonic settings but these diagrams tend to group the ocean floor basalts (OFB) under one class i.e., as mid-oceanic ridge basalts (MORB). Hence, a method is specifically needed to identify the OFB as normal (N-MORB), enriched (E MORB) and ocean island basalts (OIB). We have applied Artificial Neural Network (ANN) technique as a supervised Learning Vector Quantisation (LVQ) to identify the inherent geochemical signatures present in the Central Indian Ocean Basin (CIOB) basalts. A range of N-MORB, E-MORB and OIB dataset was used for training and testing of the network. Although the identification of the characters as N MORB, E-MORB and OIB is completely dependent upon the training data set for the LVQ, but to a significant extent this method is found to be successful in identifying the characters within the CIOB basalts. The study helped to geochemically delineate the CIOB basalts as N-MORB with perceptible imprints of E MORB and OIB characteristics in the form of moderately enriched rare earth and incompatible elements. Apart from the fact that the magmatic processes are difficult to be deciphered, the architecture performs satisfactorily. PMID- 20028565 TI - The relation of metabolic syndrome according to five definitions to cardiovascular risk factors--a population-based study. AB - BACKGROUND: Although National Cholesterol Education Program (NCEP), International Diabetes Federation (IDF), American Heart Association and National Heart, Lung and Blood Institute (AHA/NHLBI), World Health Organization (WHO), and the European Group for the Study of Insulin Resistance (EGIR) definitions of metabolic syndrome (MetS) have been commonly used by studies, little is known about agreement among these five definitions. We examined the agreement among these five definitions and explored their relationship with risk factors of cardiovascular disease in a Taiwan population. METHODS: A total of 1305 subjects aged 40 years and over in Taiwan were analyzed. Biomedical markers and anthropometric indices were measured. Agreement among definitions was determined by the kappa statistic. Logistic regression models were fit to estimate the odds of a high cardiovascular risk group for five definitions of MetS. RESULTS: The agreement among the NCEP, IDF, and AHA/NHLBI definitions was from substantial to very good, and agreement between the WHO and EGIR definitions was also substantial. All MetS definitions were significantly associated prevalence of microalbuminuria, elevated highly sensitive CRP (hs-CRP), and arterial stiffness only in women. In men, MetS by NCEP and AHA/NHLBI was associated with elevated level of hs-CRP and arterial stiffness. MetS by WHO and EGIR were significantly associated with microalbuminuria. And MetS by WHO was the only MetS definition that significantly associated with prevalence of arterial stiffness (OR: 2.75, 95% CI: 1.22-6.19). CONCLUSIONS: The associations of these five definitions with cardiovascular risk factors were similar in women, and it was evident that the five definitions performed better in women than in men, with higher ORs observed in relation to arterial stiffness, elevated hs-CRP, and higher Framingham risk scores. PMID- 20028566 TI - Effectiveness of adjuvant radiotherapy in patients with oropharyngeal and floor of mouth squamous cell carcinoma and concomitant histological verification of singular ipsilateral cervical lymph node metastasis (pN1-state)--a prospective multicenter randomized controlled clinical trial using a comprehensive cohort design. AB - BACKGROUND: Modern radiotherapy plays an important role in therapy of advanced head and neck carcinomas. However, no clinical studies have been published addressing the effectiveness of postoperative radiotherapy in patients with small tumor (pT1, pT2) and concomitant ipsilateral metastasis of a single lymph node (pN1), which would provide a basis for a general treatment recommendation. METHODS/DESIGN: The present study is a non-blinded, prospective, multi-center randomized controlled trial (RCT). As the primary clinical endpoint, overall survival in patients receiving postoperative radiation therapy vs. patients without adjuvant therapy following curative intended surgery is compared. The aim of the study is to enroll 560 adult males and females for 1:1 randomization to one of the two treatment arms (irradiation/no irradiation). Since patients with small tumor (T1/T2) but singular lymph node metastasis are rare and the amount of patients consenting to randomization is not predictable in advance, all patients rejecting randomization will be treated as preferred and enrolled in a prospective observational study (comprehensive cohort design) after giving informed consent. This observational part of the trial will be performed with maximum consistency to the treatment and observation protocol of the RCT. Because the impact of patient preference for a certain treatment option is not calculable, parallel design of RCT and observational study may provide a maximum of evidence and efficacy for evaluation of treatment outcome. Secondary clinical endpoints are as follows: incidence and time to tumor relapse (locoregional relapse, lymph node involvement and distant metastatic spread), Quality of life as reported by EORTC (QLQ-C30 with H&N 35 module), and time from operation to orofacial rehabilitation. All tumors represent a homogeneous clinical state and therefore additional investigation of protein expression levels within resection specimen may serve for establishment of surrogate parameters of patient outcome. CONCLUSION: The inherent challenges of a rare clinical condition (pN1) and two substantially different therapy arms would limit the practicality of a classical randomized study. The concept of a Comprehensive Cohort Design combines the preference of a randomized study, with the option of careful data interpretation within an observational study. TRIAL REGISTRATION: ClinicalTrials.gov: NCT00964977. PMID- 20028567 TI - Cancer risk in MLH1, MSH2 and MSH6 mutation carriers; different risk profiles may influence clinical management. AB - BACKGROUND: Lynch syndrome (LS) is associated with a high risk for colorectal cancer (CRC) and extracolonic malignancies, such as endometrial carcinoma (EC). The risk is dependent of the affected mismatch repair gene. The aim of the present study was to calculate the cumulative risk of LS related cancers in proven MLH1, MSH2 and MSH6 mutation carriers. METHODS: The studypopulation consisted out of 67 proven LS families. Clinical information including mutation status and tumour diagnosis was collected. Cumulative risks were calculated and compared using Kaplan Meier survival analysis. RESULTS: MSH6 mutation carriers, both males and females had the lowest risk for developing CRC at age 70 years, 54% and 30% respectively and the age of onset was delayed by 3-5 years in males. With respect to endometrial carcinoma, female MSH6 mutation carriers had the highest risk at age 70 years (61%) compared to MLH1 (25%) and MSH2 (49%). Also, the age of EC onset was delayed by 5-10 years in comparison with MLH1 and MSH2. CONCLUSIONS: Although the cumulative lifetime risk of LS related cancer is similar, MLH1, MSH2 and MSH6 mutations seem to cause distinguishable cancer risk profiles. Female MSH6 mutation carriers have a lower CRC risk and a higher risk for developing endometrial carcinoma. As a consequence, surveillance colonoscopy starting at age 30 years instead of 20-25 years is more suitable. Also, prophylactic hysterectomy may be more indicated in female MSH6 mutation carriers compared to MLH1 and MSH2 mutation carriers. PMID- 20028568 TI - Renal malignancy and inferior vena cava thrombus. PMID- 20028569 TI - Intravesical therapy for urothelial carcinoma of the urinary bladder: a critical review. AB - The management of non-muscle-invasive urothelial carcinoma of the bladder (UCB) is a challenge for physicians and patients alike. This is largely due to the heterogeneous natural history of this disease, in which tumors range from indolent to rapidly progressive and eventually fatal. Moreover, the high rate of recurrence and progression cause significant morbidity, expense, and detriment to quality of life. The advent of effective and safe intravesical therapies has improved the management of non-muscle-invasive UCB. Nevertheless, despite over 30 years of research and clinical experience, the mechanism, risks, benefits, and optimal regimens and treatment algorithms remain unclear. Although immunotherapy with bacillus Calmette-Guerin (BCG) has been the mainstay of intravesical treatment and represents a significant advance in the interaction of immunology and oncology, its clinical effectiveness is accompanied by a wide range of adverse events. Here, we review the literature on intravesical immunotherapy and chemotherapy with the aim of evaluating the clinical utility of the different treatments and providing recommendations. Many studies over the years have compared efficacy and toxicities of different agents and regimens, and certain conclusions are now well supported by high-level evidence. Future perspectives and promising advances in drug development are discussed and areas of improvement are identified in order to promote better cancer control and decrease the rate and severity of side-effects. PMID- 20028570 TI - Tumor thrombus involving the inferior vena cava in renal malignancy: is there a difference in clinical presentation and outcome among right and left side tumors? AB - PURPOSE: Renal cell carcinoma (RCC) has a propensity to propagate into the renal vein and inferior vena cava (IVC). Due to inherent differences in the venous anatomy of the right and left kidneys, tumor thrombus involvement of IVC may vary. The aim of this study is to compare clinical presentation and outcome of right vs. left RCC with IVC thrombus. MATERIALS AND METHODS: Patients who underwent radical nephrectomy and IVC thrombectomy between 1997 and 2008 were identified. All relevant data were collected and analyzed. RESULTS: Eight-seven patients were included. Sixty patients (69%) had a right sided tumor. Mean tumor size was 10.2 (+/- 4) cm and was not significantly different on either side. Fifty-six percent of right sided tumors had level-III (intra-hepatic) or higher tumor thrombus, while 22% of left sided tumors had similar level thrombus extension (p < 0.0001). Nearly 50% of left sided tumors showed level-I thrombus compared to 10% of right side tumors. A comparison of age, estimated blood loss and transfusion rate was not significantly different. The recurrence free (p = 0.9) and disease specific survival (p = 0.4) were not significantly different between the right and left side tumors with IVC thrombus. CONCLUSION: A level-III IVC tumor thrombus is more frequently seen with a right side tumor. However, clinical and operative characteristics among the left and right sided tumors with IVC thrombus were not different. More significantly, recurrence rate and survival did not differ with the laterality of the tumor. PMID- 20028571 TI - Metabolic investigation of patients with staghorn calculus: is it necessary? AB - OBJECTIVE: To evaluate the prevalence of metabolic disorders in patients with staghorn calculi treated at the Regional Center of Lithiasis Metabolic Studies in central region of Sao Paulo State, Brazil. MATERIALS AND METHODS: Between February 2000 and February 2008, 630 patients with urinary calculi were evaluated in the lithiasis outpatient clinic. Thirty-seven of them had staghorn calculi (35 women and 2 men). The inclusion criteria for the metabolic investigation included the absence of urological manipulation 30 days before the examination, negative urine culture and creatinine clearance > 60 mL/min. The protocol for metabolic investigation consisted of qualitative search for cystinuria. Two non-consecutive 24-hour urine samples collected to measure calcium, phosphorus, uric acid, sodium, potassium, magnesium, oxalate and citrate, and serum calcium levels, phosphorus, uric acid, sodium, potassium, magnesium, chloride, parathormone and urine pH. RESULTS: Among patients with lithiasis, 5.9% (37/630) had staghorn calculus and in 48.6% (18/37) were diagnosed with urinary infection. The females were predominant for 94.5% of cases. The calculi were unilateral in 31 of cases and bilateral in six. Metabolic abnormalities were found in 68.2% of patients with hypercalciuria (64.2%) and hypocitraturia (53.3%) being the most common disorders. CONCLUSIONS: The presence of metabolic disorders in nearly 70% of patients with staghorn calculus reinforces the necessity for evaluation of these patients. The diagnosis and treatment of identified metabolic abnormalities can contribute to the prevention of recurrent staghorn calculi. PMID- 20028572 TI - UroVysion testing can lead to early identification of intravesical therapy failure in patients with high risk non-muscle invasive bladder cancer. AB - PURPOSE: In this study, we investigated the ability of UroVysion to assess response to intravesical therapy in patients with high risk superficial bladder tumors. MATERIALS AND METHODS: We performed a retrospective review of patients undergoing intravesical therapy for high risk superficial bladder tumors. Urine specimens were collected for UroVysion analysis before and immediately after a course of intravesical therapy. Cytology and cystoscopy were performed six weeks after treatment, using either a positive cytology or visible abnormality on cystoscopy as a prompt for biopsy. The operating characteristics of the UroVysion test were then determined. RESULTS: 41 patients were identified in whom 47 cycles of induction and 41 cycles of maintenance intravesical therapy were given during the study period. This yielded a total of 88 treatment and evaluation cycles. Median follow-up was 9 months per induction (range 1-21 months) and 13 months per patient (range 1-25 months). A total of 133 urine samples were collected for UroVysionTM of which 40 were positive. Based upon standard clinical evaluation, 41 biopsies were performed which detected 20 recurrences. UroVysionTM testing performed immediately upon completion of therapy for the 41 patients undergoing biopsy yielded a sensitivity, specificity, and accuracy of 85%, 61%, and 71%. CONCLUSIONS: The use of UroVysionTM following intravesical therapy for high-risk superficial bladder tumors helps to identify patients at high risk of refractory or recurrent disease who should undergo immediate biopsy under anesthesia. PMID- 20028573 TI - Altered male physiologic function after surgery for prostate cancer: couple perspective. AB - PURPOSE: Both the diagnosis of prostate cancer (PCa) and the physiologic outcomes of surgical treatment impact the male's psychological sphere. However, current research advocates a refocusing of outcomes directed to the PCa "couple". Herein we acquire insight into perspective and concordance regarding male physiological function from the standpoint of a couple recovering from PCa surgery. MATERIALS AND METHODS: Couples whereby the male partner had undergone primary surgical treatment for PCa were mailed a Retrospective Sexual Survey (RSS) packet consisting of male and female partner questionnaires. RSS questions surveyed physiological changes in libido, foreplay, erection and arousal, orgasm and ejaculation in addition to perceived psychological impact. Patients' and partners' scores were evaluated to determine the concordance of both individual items as well as domain sums. RESULTS: Twenty-eight couples completed the questionnaires. Only about 40% of men and women were happy with their levels of sexual interest with 82% concordance. Urine loss during orgasm was reported by 43% of men; the majority of participants were bothered by it. Ejaculation changes were observed by 96% of men (concordance 96%) with most reporting anejaculation. A change in orgasm experience was noted by 86% of men (and 36% of their female partners, p < 0.0001). Despite the change, the majority of men and women reported being satisfied with their ability to climax. CONCLUSION: Our results indicate that patients and their female partners may interpret differently the same physiological outcomes of PCa surgery. This information could be useful to better counsel the PCa couple and help patients and partners adjust after surgery. PMID- 20028574 TI - Transurethral resection of the prostate for the treatment of lower urinary tract symptoms related to benign prostatic hyperplasia: how much should be resected? AB - OBJECTIVE: To assess the impact of the percent of resected tissue on the improvement of urinary symptoms. MATERIALS AND METHODS: The study included a prospective analysis of 88 men with benign prostatic hyperplasia. Patients were divided in three groups according to the percent of resected tissue: Group 1 < 30%; Group 2, 30% to 50%; and Group 3, > 50%. Each patient was re-evaluated 3 months after surgery. We assessed the international prostatic symptom score, nocturia and serum prostate specific antigen levels. RESULTS: All patients presented a significant decrease on mean International Prostate System Score (IPSS) (23 to 5.9), Quality of Life (QoL) (4.9 to 1.0) and nocturia (3.2 to 1.9). Variation in the IPSS was 16.7, 16.6 and 18.4 for patients from Group 1, 2 and 3 respectively (P = 0.504). Although the three groups presented a significant decrease in QoL, patients in Group 3 presented a significantly greater decrease when compared to Group 1. Variation in QoL was 3.1, 3.9 and 4.2 for patients from Group 1, 2 and 3 respectively (p = 0.046). There was no significant difference in nocturia variation according to the percent of resected tissue (p = 0.504). Median pre and postoperative PSA value was 3.7 and 1.9 ng/mL respectively. Patients from Group 1 did not show a significant variation (p = 0.694). Blood transfusions were not required in any group. CONCLUSIONS: Resection of less than 30% of prostatic tissue seems to be sufficient to alleviate lower urinary tract symptoms related to benign prostate hyperplasia. However, these patients may not show a significant decrease in serum PSA level. PMID- 20028575 TI - Imaging of the acute scrotum: is there a place for contrast-enhanced ultrasonography? AB - PURPOSE: To present and evaluate the findings of contrast-enhanced ultrasonography (CEUS) in typical cases of acute painful scrotum. MATERIALS AND METHODS: Nineteen patients aged from 19 to 61 years old were included in the study. All patients underwent grey-scale and color Doppler ultrasonography (US) of the scrotum, followed by imaging after i.v. administration of 2.4 mL of a second generation ultrasound contrast agent (microbubbles of sulphur hexafluoride). ? dedicated, contrast-sensitive technique was used (Contrast Tissue Imaging - CnTI). The diagnosis was confirmed surgically in 6 cases while in the remaining 8 cases it was based on the combination of clinical, imaging and laboratory findings. RESULTS: The final diagnosis was testicular torsion (n = 4), epididymitis (n = 2, one of the cases complicated by abscess), testicular abscess (n = 1), scrotal abscess (n = 1), testicular trauma of varying severity (n = 6). Five out of 19 cases were true negatives: neither clinical examination nor laboratory tests revealed any pathology. CEUS showed complete lack of enhancement in all cases of torsion, permitting a rapid and definitive diagnosis. In the cases of infection complicated by abscesses, CEUS delineated the lesions much better than the combination of B-mode/Color Doppler US. The severely traumatized testicles showed minimal, inhomogeneous or patchy enhancement, while cases of minor trauma showed no significant enhancement defects. Hematomas were presented as non-enhancing lesions. CONCLUSION: Generally, there was no advantage over Doppler US as has been previously shown. However, CEUS can be used supplementary to traditional Doppler US in the investigation of blunt testicular trauma especially when there is uncertainty in diagnosis after appropriate clinical and radiographic evaluations occurs. Further studies are required to clearly define the indications of this method. PMID- 20028576 TI - Transobturator male sling TOMS for the treatment of stress post-prostatectomy incontinence, initial experience and results with one year's experience. AB - PURPOSE: Post-prostatectomy incontinence remains a problem, even in minor or moderate degrees. In order to minimize surgical morbidity and costs, sling procedures have been proposed. The authors have developed a new transobturator male sling procedure and report their results after one-year experience. MATERIALS AND METHODS: A prospective multicenter study was conducted in 50 patients with minor or moderate post-prostatectomy incontinence. Evaluation of TOMS two arms bulbar sling was based on clinical form assessment, The International Consultation on Incontinence Questionnaire (ICIQ) and short-form (SF) 36 questionnaire pre and postoperatively and at 3, 6, 9 and 12 months. RESULTS: The surgical procedure was considered easy to perform and no post surgery complication was reported except for one retention. The median number of pads per day decreased significantly from 2 pads before surgery (95% CI: 2 - 3) to 1 during the follow-up period (95% CI: 0 - 2 at 360 days), and at 3 months patients using none or one pad per day were 30% and 32% respectively. The SF 36 continence and quality of life score improved from a median of 100 (95% CI: 83 - 133) to 300 (95% CI: 167 - 375), and the median ICIQ incontinence and quality of life score decreased from 15 (95% CI: 14 - 16) to 8 (95% CI: 5 - 12) one year after surgery. CONCLUSION: The transobturator perineal male sling TOMS is an attractive simple sling technique for moderate or minor post-prostatectomy stress incontinence and offers an improvement in the quality of life. PMID- 20028577 TI - Alfuzosin hydrochloride transdermal films: evaluation of physicochemical, in vitro human cadaver skin permeation and thermodynamic parameters. AB - PURPOSE: The main objective of the investigation was to develop a transdermal therapeutic system for alfuzosin hydrochloride and to study the effects of polymeric system and loading dose on the in vitro skin permeation pattern. MATERIALS AND METHODS: Principles of experimental design have been exploited to develop the dosage form. Ratio of ethyl cellulose (EC) and polyvinyl pyrrolidone (PVP) and loading dose were selected as independent variables and their influence on the cumulative amount of alfuzosin hydrochloride permeated per cm2 of human cadaver skin at 24 h (Q24), permeation flux (J) and steady state permeability coefficient (PSS) were studied using experimental design. Various physicochemical parameters of the transdermal films were also evaluated. Activation energy for in vitro transdermal permeation has been estimated. RESULTS: Ratio of EC and PVP was found to be the main influential factor for all the dependent variables studied. Drug loading dose was also found to influence the dependent variables but to a lesser extent. Physicochemical parameters of the prepared films were evaluated and found satisfactory. Activation energy for alfuzosin permeation has also been estimated and reported. CONCLUSION: The therapeutic system was found to be dermatologically non-irritant and hence, a therapeutically effective amount of alfuzosin hydrochloride can be delivered via a transdermal route. PMID- 20028598 TI - Rethinking the acquisition of relative clauses in Italian: towards a grammatically based account. AB - In a number of studies, the acquisition of restrictive relative clauses (RCs) shows contrasting findings regarding comprehension and production, with the former usually delayed up to the age of five. As previously claimed in the literature, we suggest that this delay is a task artifact and we present a new procedure for the assessment of restrictive RCs. Data from three- to seven-year old Italian children were collected and results show that children understand object RCs with preverbal subject in an adult-like manner at four years of age, but some of the three-year-old children were already above chance. Subject relatives show at ceiling performance from three years of age. We consider our results as evidence of continuity between early and adult competence grammars. Children's non-target responses are interpreted as grammatical options exploited by an immature performance system. PMID- 20028599 TI - Concord grape juice supplementation improves memory function in older adults with mild cognitive impairment. AB - Concord grape juice contains polyphenol compounds, which have antioxidant and anti-inflammatory properties and influence neuronal signalling. Concord grape juice supplementation has been shown to reduce inflammation, blood pressure and vascular pathology in individuals with CVD, and consumption of such flavonoid containing foods is associated with a reduced risk for dementia. In addition, preliminary animal data have indicated improvement in memory and motor function with grape juice supplementation, suggesting potential for cognitive benefit in ageing humans. In this initial investigation of neurocognitive effects, we enrolled twelve older adults with memory decline but not dementia in a randomised, placebo-controlled, double-blind trial with Concord grape juice supplementation for 12 weeks. We observed significant improvement in a measure of verbal learning and non-significant enhancement of verbal and spatial recall. There was no appreciable effect of the intervention on depressive symptoms and no effect on weight or waist circumference. A small increase in fasting insulin was observed for those consuming grape juice. These preliminary findings suggest that supplementation with Concord grape juice may enhance cognitive function for older adults with early memory decline and establish a basis for more comprehensive investigations to evaluate potential benefit and assess mechanisms of action. PMID- 20028600 TI - Peanut protein reduces body protein mass and alters skeletal muscle contractile properties and lipid metabolism in rats. AB - It is well known that diets high in nuts or peanuts favourably affect plasma lipid concentrations. However, few studies have examined the effects of nut and peanut protein (PP) on body composition and skeletal muscle properties. The present study was aimed at evaluating the effect of dietary PP compared with two animal proteins, casein (C) and cod protein (CP) on body composition, skeletal muscle contractile properties and lipid metabolism in rats. Thirty-two male rats were assigned to one of the following four diets containing either C, CP, PP or C+peanut protein (CPP, 50:50) mixture. After 28 d of ad libitum feeding and after 12-h fast, blood, liver and muscle were collected for measurements of plasma and hepatic cholesterol and TAG, plasma glucose and insulin and contractile properties. Rats fed with the low-quality protein, PP, had lower body weight gain, body protein mass, soleus mass and liver weight than those fed with the high-quality dietary proteins, C and CP. PP also caused a deficit in contractile properties in soleus. Likewise, PP increased plasma cholesterol and body fat mass compared with CP. However, these elevations were accompanied with increased hepatic TAG concentrations and lowered intestinal fat excretion. These results show that PP intake alters body composition by reducing skeletal muscle mass and liver weight as well as muscle contractility and lipid metabolism. Adding a complete protein such as C might partially counteract these adverse effects. PMID- 20028601 TI - Trends in food and nutritional intakes of French adults from 1999 to 2007: results from the INCA surveys. AB - Two independent cross-sectional dietary surveys (the Individual and National Food Consumption Surveys, INCA), performed in 1998-99 (INCA1) and in 2006-07 (INCA2) on nationally representative samples of French people, were used to analyse trends in the dietary habits and nutritional intake of French adults. Food consumption was recorded through 7-d dietary records, and nutritional intakes were assessed using the French food composition database. After exclusion of under-reporters, analyses were performed on 3267 adults, aged 18-79 years: 1345 from INCA1 and 1922 from INCA2. The trends highlighted over the 8-year period showed a decrease in consumption of dairy products, meat, bread, potatoes, pastries/croissant-like pastries/cakes/biscuits and sugar/confectionery. In contrast, the consumption of fruits and vegetables, rice, ice cream and chocolate increased. Other food groups, like fish and snacking foods, remained stable. Food choices were mostly age specific. These age differences remained consistent over the years and underlined two opposite dietary trends: a 'traditional' one mainly followed by the elderly, and a 'snacking and convenience' one mainly adopted by young adults. The overall trends in food consumption did not influence the mean energy intake, but did slightly modify the contribution of each macronutrient to energy intake. These repeated surveys highlighted the fact that trends in French food habits have moved towards an average European diet at the crossroads between Mediterranean and Northern diets, and that food consumption changes impacted, to a lesser extent, nutritional intake. PMID- 20028602 TI - Native and microwaved bean and pea starch preparations: physiological effects on the intestinal ecosystem, caecal tissue and serum lipids in rats. AB - Dietary beans and peas provide fibre, resistant starch and other nutrients that are often lacking in the human diet. The influence of native starches of beans and peas (and microwaved preparations) on N utilisation, biochemical indices in blood serum and caecal ecosystem state (SCFA, bacterial enzymes, micro-organisms) was studied in vivo. The native pea starch contained more resistant starch compared with its bean counterpart (31 v. 17 %); however, processing decreased these amounts to 25 v. 10 %. N digestibility was found to decrease considerably in all experimental groups. A considerable reduction was observed in glucose and total cholesterol concentration in rat blood serum as a result of feeding both dietary legume starch preparations under microwave treatment. This indicates that starch of bean origin activated glycolytic bacterial enzymes; however, all the analysed starches were found to reduce the activity of beta-glucuronidase. In addition, both dietary bean starches significantly induced the formation of SCFA in the caecal digesta. As compared with the control group, a significant decrease in the pH of caecal and colonic digesta was demonstrated for both bean starch preparations. In comparison with the diet with native pea starch, its microwaved preparation reduced the concentrations of acetic, butyric and propionic acids among caecal SCFA and increased the pH of caecal and colonic digesta. The atherogenic index was significantly lower in rats fed microwaved pea starch. All investigated starch preparations increased the population of Bifidobacterium spp. in caecal digesta, but were also good substrates for opportunistic Enterococcus or Escherichia coli. PMID- 20028603 TI - Resolution of hyperacusis associated with depression, following lithium administration and directive counselling. AB - INTRODUCTION: Hyperacusis may be described as a decreased threshold for discomfort from sound. It may have a serious impact on an individual's lifestyle, as a result of decreased sociability and inability to spend time with family and friends due to intolerance of sound. SUBJECT: We present a case of hyperacusis associated with depression, with no other audiological abnormality. The patient reported greater difficulty during the depressive episode. METHOD: Audiological investigation was undertaken, followed by counselling and medication for depression. RESULTS AND CONCLUSION: The pathophysiological mechanism of hyperacusis in depression is discussed. Outcomes following treatment with directive counselling and lithium (an antidepressant) are described. Research indicates that both hyperacusis and depression can be caused by hypoactivity of 5 hydroxytryptamine (also known as serotonin) in the brain. The patient reported improvement which cannot be explained solely on the basis of counselling. Enhancement of serotonin activity due to lithium, prescribed for depression, may also play a role in alleviation of hyperacusis. PMID- 20028604 TI - An unusual case of cotton wool use to improve hearing. AB - OBJECTIVES: To report an unusual observation whereby hearing was improved by insertion of cotton wool onto a retracted tympanic membrane, and to explore potential explanations for this. CASE REPORT: We report the case of a 56-year-old woman with a severe mixed hearing loss who was noted to have a significant improvement in her hearing using a technique shown to her many years ago by a Russian physician. Upon application of a moist piece of cotton wool, gains of up to 40 dB were observed. This was associated with a significant subjective improvement in hearing. We discuss potential mechanisms for this effect, including amplification, a baffle effect and phase differences. CONCLUSION: The mechanism of the hearing improvement observed in this case is unknown, but is likely to be related to sound amplification from the relatively large surface area of the cotton wool to the smaller oval window. PMID- 20028605 TI - Oral voriconazole for invasive fungal skull base infection. AB - BACKGROUND: Intravenous amphotericin or intravenous voriconazole, both followed by oral voriconazole, have previously been given to treat invasive aspergillosis of the skull base. CASE REPORT: Exclusively oral voriconazole was used in an immunocompetent patient with biopsy-proven, invasive aspergillosis. She had a large, erosive lesion extending from the central skull base to the right orbit and ethmoid sinus, and displacing the right internal carotid artery. After four months of oral treatment as an out-patient, a repeated computed tomography scan showed a fully treated infection with post-infectious changes only, and treatment was terminated. Two years later, there had been no recurrence. CONCLUSION: Substantial cost savings were made by using exclusively oral treatment, compared with the use of intravenous voriconazole or amphotericin, or a switch strategy. PMID- 20028606 TI - Isolated fungus ball mimicking mucocele or frontal sinus tumour: a diagnostic pitfall. AB - OBJECTIVE: To demonstrate the clinical, radiological and diagnostic pitfalls of managing an isolated frontal sinus fungus ball, and to compare with the literature. MATERIAL AND METHODS: Retrospective analysis of two cases and literature review. RESULTS: Isolated frontal sinus fungus ball is a rare cause of frontal sinus disease. We present two cases of isolated frontal sinus fungus ball which pre-operatively were suspected to be either a tumour or a mucocele. In both cases, cheesy, clay-like material was found intra-operatively within the frontal sinus, suggesting a fungus ball. Effective treatment included surgical debridement via an exclusively endoscopic or an external approach, variously. Final histopathological and mycological analysis revealed Aspergillus fumigatus. A literature review revealed 20 reported cases of isolated frontal sinus fungus ball, confirming the low prevalence of the disease. CONCLUSIONS: Frontal sinus fungus ball should be considered in the differential diagnosis of chronic, nonspecific forehead symptoms. To evaluate the underlying disease, computed tomography scans should first be performed, followed by magnetic resonance imaging if malignancy is suspected. It is essential to be aware of the possibility of an atypical fungus ball appearance on computed tomography and magnetic resonance imaging scans. If bony destruction and calcification coexist on radiological images, then endoscopic biopsy is an indispensable part of the diagnostic procedure, and should be performed to collect material for both histological and mycological analysis, and to aid surgical planning. In cases of sinus fungus ball, an endoscopic approach for biopsy may be curative. PMID- 20028608 TI - Cerebral malaria: why experimental murine models are required to understand the pathogenesis of disease. AB - Cerebral malaria is a life-threatening complication of malaria infection. The pathogenesis of cerebral malaria is poorly defined and progress in understanding the condition is severely hampered by the inability to study in detail, ante mortem, the parasitological and immunological events within the brain that lead to the onset of clinical symptoms. Experimental murine models have been used to investigate the sequence of events that lead to cerebral malaria, but there is significant debate on the merits of these models and whether their study is relevant to human disease. Here we review the current understanding of the parasitological and immunological events leading to human and experimental cerebral malaria, and explain why we believe that studies with experimental models of CM are crucial to define the pathogenesis of the condition. PMID- 20028607 TI - Multilocus phylogenetic analyses reveal that habitat selection drives the speciation of Didymozoidae (Digenea) parasitizing Pacific and Atlantic bluefin tunas. AB - Parasite communities of wild and reared bluefin tuna display remarkable diversity. Among these, the most prevalent and abundant are the Didymozoidae (Monticelli, 1888) (Trematoda, Digenea), considered one of the most taxonomically complex digenean families. The aim of this study was to evaluate phylogenetic structure of Didymozoidae occurring in Pacific (Thunnus orientalis) and Atlantic bluefin tuna (T. thynnus) in order to increase our knowledge of didymozoid zoogeography and identify species that could successfully be employed as biological tags for stock assessment studies. For the present analyses we used 2 nuclear ribosomal DNA loci, part of the 28S gene and the second internal transcribed spacer (ITS-2) as well as a portion of the mitochondrial cytochrome c oxidase subunit 1 gene (cox1). In most parasitic groups, morphology is the primary factor in the structuring of phylogenetic relationships. In rare examples, however, habitat has been suggested as a primary factor affecting parasite evolution. During their evolution, didymozoids have spread and inhabited a remarkable number of different sites in their hosts, colonizing exterior as well as strictly interior niches. Our data suggest that habitat selection has been the leading force in shaping didymozoid phylogenetic relationships. For 2 didymozoid species (D. wedli and D. palati), cox1 sequences indicate intraspecific differences between Mexican and Adriatic populations. PMID- 20028609 TI - Concurrent nematode infection and pregnancy induce physiological responses that impair linear growth in the murine foetus. AB - This study examined concurrent stresses of nematode infection and pregnancy using pregnant and non-pregnant CD1 mice infected 3 times with 0, 50 or 100 Heligmosomoides bakeri larvae. Physiological, energetic, immunological and skeletal responses were measured in maternal and foetal compartments. Resting metabolic rate (RMR) was elevated by pregnancy, but not by the trickle infection. Energy demands during pregnancy were met through increased food intake and fat utilization whereas mice lowered their body temperature during infection. Both infection and pregnancy increased visceral organ mass and both altered regional bone area and mineralization. During pregnancy, lumbar mineralization was lower but femur area and mineralization were higher. On the other hand, infection lowered maternal femur bone area and this was associated with higher IFN-gamma in maternal serum of heavily infected pregnant mice. Infection also reduced foetal crown-rump length which was associated with higher amniotic fluid IL-1 beta. PMID- 20028610 TI - Trypanosomiasis and the brain. AB - Neurological involvement following trypanosome infection has been recognised for over a century. However, there are still many unanswered questions concerning the mechanisms used by the parasite to gain entry to the CNS and the pathogenesis of the resulting neuroinflammatory reaction. There is a paucity of material from human cases of the disease therefore the majority of current research relies on the use of animal models of trypanosome infection. This review reports contemporary knowledge, from both animal models and human samples, regarding parasite invasion of the CNS and the neuropathological changes that accompany trypanosome infection and disease progression. The effects of trypanosomes on the blood-brain barrier are discussed and possible key molecules in parasite penetration of the barrier highlighted. Changes in the balance of CNS cytokines and chemokines are also described. The article closes by summarising the effects of trypanosome infection on the circadian sleep-wake cycle, and sleep structure, in relation to neuroinflammation and parasite location within the CNS. Although a great deal of progress has been made in recent years, the advent and application of sophisticated analysis techniques, to decipher the complexities of HAT pathogenesis, herald an exciting and rewarding period for advances in trypanosome research. PMID- 20028611 TI - Acetate and succinate production in amoebae, helminths, diplomonads, trichomonads and trypanosomatids: common and diverse metabolic strategies used by parasitic lower eukaryotes. AB - Parasites that often grow anaerobically in their hosts have adopted a fermentative strategy relying on the production of partially oxidized end products, including lactate, glycerol, ethanol, succinate and acetate. This review focuses on recent progress in understanding acetate production in protist parasites, such as amoebae, diplomonads, trichomonads, trypanosomatids and in the metazoan parasites helminths, as well as the succinate production pathway(s) present in some of them. We also describe the unconventional organisation of the tricarboxylic acid cycle associated with the fermentative strategy adopted by the procyclic trypanosomes, which may resemble the probable structure of the primordial TCA cycle in prokaryotes. PMID- 20028612 TI - A mathematical model of the indirect effects of rotavirus vaccination. AB - Rotavirus (RV) infections progressively confer natural immunity against subsequent infection. Similarly to natural infection, vaccination with a live attenuated vaccine potentially reduces RV transmission and induces herd protection. A mathematical transmission model was developed to project the impact of a vaccination programme on the incidence of RV infection and disease for five countries in the European Union. With vaccination coverage rates of 70%, 90% and 95% the model predicted that, in addition to the direct effect of vaccination, herd protection induced a reduction in RV-related gastroenteritis (GE) incidence of 25%, 22% and 20%, respectively, for RV-GE of any severity, and of 19%, 15%, and 13%, respectively, for moderate-to-severe RV-GE, 5 years after implementation of a vaccination programme. PMID- 20028613 TI - Bevacizumab, sorafenib tosylate, sunitinib and temsirolimus for renal cell carcinoma: a systematic review and economic evaluation. AB - OBJECTIVES: To assess the clinical effectiveness and cost-effectiveness of bevacizumab, combined with interferon (IFN), sorafenib tosylate, sunitinib and temsirolimus in the treatment of people with advanced and/or metastatic renal cell carcinoma (RCC). DATA SOURCES: Electronic databases, including MEDLINE, EMBASE and the Cochrane Library, were searched up to September/October 2007 (and again in February 2008). REVIEW METHODS: Systematic reviews and randomised clinical trials comparing any of the interventions with any of the comparators in participants with advanced and/or metastatic RCC were included, also phase II studies and conference abstracts if there was sufficient detail to adequately assess quality. Results were synthesised narratively and a decision-analytic Markov-type model was developed to simulate disease progression and estimate the cost-effectiveness of the interventions under consideration. RESULTS: A total of 888 titles and abstracts were retrieved in the clinical effectiveness review, including reports of eight clinical trials. Treatment with bevacizumab plus IFN or sunitinib had clinically relevant and statistically significant advantages over treatment with IFN alone, in terms of progression-free survival and tumour response, doubling median progression-free survival from approximately 5 months to 10 months. Temsirolimus had similar advantages over treatment with IFN in terms of progression-free and overall survival, increasing median overall survival from 7.3 to 10.9 months [hazard ratio (HR) 0.73; 95% confidence interval (CI) 0.58 to 0.92)], as did sorafenib in comparison with best supportive care in terms of overall survival, progression-free survival and tumour response, with a doubling of progression-free survival (HR 0.51; 95% CI 0.43 to 0.60). However, the last was associated with an increased frequency of hypertension and hand-foot skin reaction compared with placebo. No fully published economic evaluations of any of the interventions could be located. However, estimates from the PenTAG model suggested that none of the interventions would be considered cost-effective at a willingness-to-pay threshold of 30,000 pounds per quality-adjusted life-year (QALY). Estimates of cost per QALY ranged from 71,462 pounds for sunitinib to 171,301 pounds for bevacizumab plus IFN. Although there are many similarities in the methodology and structural assumptions employed by PenTAG and the manufacturers of the interventions, in all cases the cost-effectiveness estimates from the PenTAG model were higher than those presented in the manufacturers' submissions. Cost-effectiveness estimates were particularly sensitive to variations in the estimates of treatment effectiveness, drug pricing (including dose intensity data), and health-state utility input parameters. CONCLUSIONS: Treatment with bevacizumab plus IFN and sunitinib has clinically relevant and statistically significant advantages over treatment with IFN alone in patients with metastatic RCC. In people with three of six risk factors for poor prognosis, temsirolimus had clinically relevant advantages over treatment with IFN, and sorafenib tosylate was superior to best supportive care as second-line therapy. The frequency of adverse events associated with bevacizumab plus IFN, sunitinib and temsirolimus was comparable with that seen with IFN, although the adverse event profile is different. Treatment with sorafenib was associated with a significantly increased frequency of hypertension and hand-foot syndrome. Estimates from the PenTAG model suggested that none of the interventions would be considered cost-effective at a willingness-to-pay threshold of 30,000 pounds per QALY. PMID- 20028614 TI - Reproductive phenology of transgenic Brassica napus cultivars: Effect on intraspecific gene flow. AB - Pollen-mediated gene flow in space is well documented and isolation distances are recommended to ensure genetic purity of Brassica napus seed crops. Isolation in time could also contribute to gene flow management but has been little investigated. We assessed the effects of asynchronous and synchronous flowering on intraspecific B. napus gene flow by seeding adjacent plots of transgenic spring canola cultivars, either resistant to glyphosate or glufosinate, over a 0 4 week interval and measuring outcrossing rates and seed-set. Outcrossing rates, evaluated in the center of the first adjacent row, were reduced to the lowest level in plots flowering first when the seeding interval > 2 weeks. Increasing the time gap increased outcrossing rates in plots flowering second up to a seeding interval of two weeks. Flowers that opened during the last week of the flowering period produced fewer seed (< 10% of total seed production) and a smaller fraction of outcrossed seed (-25%). Observed time gap effects were likely caused by extraneous pollen load during the receptivity of productive seed setting early flowers. Clearly, manipulation of B. napus flowering development through staggered planting dates can contribute to gene flow management. The approach will need to be validated by additional site-years and increased isolation distances. PMID- 20028615 TI - Towards a smart biosafety regulation: The case of Kenya. AB - It is emerging that benefits of new innovations in agricultural biotechnology may not be realised without appropriate biosafety regulatory mechanisms. The Kenyan regulatory experiences related to regulation of genetically engineered (GE) agricultural activities are explored to provide some basis for defining the challenges involved in biosafety regulation. The various shortcomings in the way the Kenyan biosafety process has been implemented are consistent with the way regulatory systems have been evolving in Africa. The proposed adoption of a "smart regulation" provides the basis for a learning process through which subsequent biotechnology policy initiatives can be improved. PMID- 20028616 TI - Transgenes and national boundaries - The need for international regulation. AB - What happens when one nation cultivates a transgenic crop variety but neighboring nations do not? Using alfalfa as a case study, we argue that the potential for international transgene flow is substantial, and therefore, the need for international cooperation in regulatory decisions concerning transgenic crops is imperative. Alfalfa (Medicago sativa, L.) is the major forage crop in North America. Recently, genetically modified (GM) alfalfa received a moratorium on further cultivation in the US on the grounds that the approvals were based on inadequate environmental impact assessments. With their deep root system, symbiotic nitrogen fixation, prolific seed production and prolonged dormancy, alfalfa plants are capable of establishing self-perpetuating (feral) populations in unmanaged environments. Given what is known about alfalfa pollen dispersal, such feral populations could facilitate gene flow between GM and non-GM fields. The border between the US and Canada, particularly in farming areas, is very narrow (< 10 m wide). We surveyed along the US-Canada border and found both alfalfa fields and potentially feral alfalfa plants in the ditches along the border. Our survey results provide evidence of the possibility of cross-border transgene flow, suggesting a need for international co-operative risk assessment initiatives between the US and Canada. Such situations could occur for other crops, in other international border regions as well. PMID- 20028617 TI - Response to Bagavathiannan and Van Acker's "Transgenes and national boundaries - The need for international regulations": Biotechnology developers and regulators already consider transgene movement across national boundaries and the environmental risks posed by adventitious presence of unapproved events are overstated. AB - Bagavathiannan and Van Acker propose greater international cooperation and information sharing in risk assessment for biotechnology-derived crops because pollen- and seed-mediated gene flow across political boundaries may lead to the adventitious presence of unapproved transgenes at sites along the borders of neighboring countries. However, they fail to convince us that something is wrong with the current situation and provide no details of how it could be improved. PMID- 20028618 TI - Strategies for coexistence of GM and non-GM soy from import to feed processing. AB - Regulations 1829/2003/CE and 1830/2003/CE have allowed the placing on the European market of GM products in food and feed chains, and have defined their rules of traceability and labeling. For some supply chains, like for soy and its derived products that are used in the production of feed, manufacturers have to face both non-GM and GM production, although there are no labeling requirements for animal products derived from animals fed with GMOs. This study presents the strategies of stakeholders involved in the feed production chain to maintain concurrent production of compound feed with GM and non-GM soy products, by dealing with the coexistence between those two crops. The stakeholders include importers, traders, soy processors, feed processors and retailers. The study shows that many tools are in place to ensure and maintain the current coexistence. However, a profound harmonization of procedures and methods at a European level should be encouraged. PMID- 20028619 TI - The 10th International Symposium on the Biosafety of Genetically Modified Organisms (ISBGMO), Wellington, New Zealand, November 2008. AB - The Symposium consisted of eight sessions of oral presentations as well as various workshops and poster sessions. This report reviews the presentations in the following sessions and discusses the main conclusions and issues arising from each session: Session 1: Biosafety - experience and results Session 2: Introgression, invasion and fitness Session 3: Biotic and abiotic stress resistance Session 4: GM animals Session 5: Effects of GM crops on soil ecosystems Session 7: Biocontainment methods Session 8: Post market environmental monitoring Abstracts of the presentations in these sessions are available at: http://www.isbgmo.info/assets_/isbgmo_symposium_handbook.pdf. PMID- 20028620 TI - A new method for stability analysis of recurrent neural networks with interval time-varying delay. AB - This brief deals with the problem of stability analysis for a class of recurrent neural networks (RNNs) with a time-varying delay in a range. Both delay independent and delay-dependent conditions are derived. For the former, an augmented Lyapunov functional is constructed and the derivative of the state is retained. Since the obtained criterion realizes the decoupling of the Lyapunov function matrix and the coefficient matrix of the neural networks, it can be easily extended to handle neural networks with polytopic uncertainties. For the latter, a new type of delay-range-dependent condition is proposed using the free weighting matrix technique to obtain a tighter upper bound on the derivative of the Lyapunov-Krasovskii functional. Two examples are given to illustrate the effectiveness and the reduced conservatism of the proposed results. PMID- 20028621 TI - Robust independent component analysis by iterative maximization of the kurtosis contrast with algebraic optimal step size. AB - Independent component analysis (ICA) aims at decomposing an observed random vector into statistically independent variables. Deflation-based implementations, such as the popular one-unit FastICA algorithm and its variants, extract the independent components one after another. A novel method for deflationary ICA, referred to as RobustICA, is put forward in this paper. This simple technique consists of performing exact line search optimization of the kurtosis contrast function. The step size leading to the global maximum of the contrast along the search direction is found among the roots of a fourth-degree polynomial. This polynomial rooting can be performed algebraically, and thus at low cost, at each iteration. Among other practical benefits, RobustICA can avoid prewhitening and deals with real- and complex-valued mixtures of possibly noncircular sources alike. The absence of prewhitening improves asymptotic performance. The algorithm is robust to local extrema and shows a very high convergence speed in terms of the computational cost required to reach a given source extraction quality, particularly for short data records. These features are demonstrated by a comparative numerical analysis on synthetic data. RobustICA's capabilities in processing real-world data involving noncircular complex strongly super-Gaussian sources are illustrated by the biomedical problem of atrial activity (AA) extraction in atrial fibrillation (AF) electrocardiograms (ECGs), where it outperforms an alternative ICA-based technique. PMID- 20028622 TI - A multiobjective simultaneous learning framework for clustering and classification. AB - Traditional pattern recognition involves two tasks: clustering learning and classification learning. Clustering result can enhance the generalization ability of classification learning, while the class information can improve the accuracy of clustering learning. Hence, both learning methods can complement each other. To fuse the advantages of both learning methods together, many existing algorithms have been developed in a sequential fusing way by first optimizing the clustering criterion and then the classification criterion associated with the obtained clustering results. However, such kind of algorithms naturally fails to achieve the simultaneous optimality for two criteria, and thus has to sacrifice either the clustering performance or the classification performance. To overcome that problem, in this paper, we present a multiobjective simultaneous learning framework (MSCC) for both clustering and classification learning. MSCC utilizes multiple objective functions to formulate the clustering and classification problems, respectively, and more importantly, it employs the Bayesian theory to make these functions all only dependent on a set of the same parameters, i.e., clustering centers which play a role of the bridge connecting the clustering and classification learning. By simultaneously optimizing the clustering centers embedded in these functions, not only the effective clustering performance but also the promising classification performance can be simultaneously attained. Furthermore, from the multiple Pareto-optimality solutions obtained in MSCC, we can get an interesting observation that there is complementarity to great extent between clustering and classification learning processes. Empirical results on both synthetic and real data sets demonstrate the effectiveness and potential of MSCC. PMID- 20028623 TI - A versatile model for packet loss visibility and its application to packet prioritization. AB - In this paper, we propose a generalized linear model for video packet loss visibility that is applicable to different group-of-picture structures. We develop the model using three subjective experiment data sets that span various encoding standards (H.264 and MPEG-2), group-of-picture structures, and decoder error concealment choices. We consider factors not only within a packet, but also in its vicinity, to account for possible temporal and spatial masking effects. We discover that the factors of scene cuts, camera motion, and reference distance are highly significant to the packet loss visibility. We apply our visibility model to packet prioritization for a video stream; when the network gets congested at an intermediate router, the router is able to decide which packets to drop such that visual quality of the video is minimally impacted. To show the effectiveness of our visibility model and its corresponding packet prioritization method, experiments are done to compare our perceptual-quality-based packet prioritization approach with existing Drop-Tail and Hint-Track-inspired cumulative-MSE-based prioritization methods. The result shows that our prioritization method produces videos of higher perceptual quality for different network conditions and group-of-picture structures. Our model was developed using data from high encoding-rate videos, and designed for high-quality video transported over a mostly reliable network; however, the experiments show the model is applicable to different encoding rates. PMID- 20028624 TI - Convex optimization for nonrigid stereo reconstruction. AB - We present a method for recovering 3-D nonrigid structure from an image pair taken with a stereo rig. More specifically, we dedicate to recover shapes of nearly inextensible deformable surfaces. In our approach, we represent the surface as a 3-D triangulated mesh and formulate the reconstruction problem as an optimization problem consisting of data terms and shape terms. The data terms are model to image keypoint correspondences which can be formulated as second-order cone programming (SOCP) constraints using L(infinity) norm. The shape terms are designed to retaining original lengths of mesh edges which are typically nonconvex constraints. We will show that this optimization problem can be turned into a sequence of SOCP feasibility problems in which the nonconvex constraints are approximated as a set of convex constraints. Thanks to the efficient SOCP solver, the reconstruction problem can then be solved reliably and efficiently. As opposed to previous methods, ours neither involves smoothness constraints nor need an initial estimation, which enables us to recover shapes of surfaces with smooth, sharp and other complex deformations from a single image pair. The robustness and accuracy of our approach are evaluated quantitatively on synthetic data and qualitatively on real data. PMID- 20028625 TI - Adaptive directional wavelet transform based on directional prefiltering. AB - This paper proposes an efficient approach for adaptive directional wavelet transform (WT) based on directional prefiltering. Although the adaptive directional WT is able to transform an image along diagonal orientations as well as traditional horizontal and vertical directions, it sacrifices computation speed for good image coding performance. We present two efficient methods to find the best transform directions by prefiltering using 2-D filter bank or 1-D directional WT along two fixed directions. The proposed direction calculation methods achieve comparable image coding performance comparing to the conventional one with less complexity. Furthermore, transform direction data of the proposed method can be used for content-based image retrieval to increase retrieval ratio. PMID- 20028626 TI - Nonlinear non-negative component analysis algorithms. AB - In this paper, general solutions for nonlinear non-negative component analysis for data representation and recognition are proposed. Motivated by a combination of the non-negative matrix factorization (NMF) algorithm and kernel theory, which has lead to a recently proposed NMF algorithm in a polynomial feature space, we propose a general framework where one can build a nonlinear non-negative component analysis method using kernels, the so-called projected gradient kernel non-negative matrix factorization (PGKNMF). In the proposed approach, arbitrary positive definite kernels can be adopted while at the same time it is ensured that the limit point of the procedure is a stationary point of the optimization problem. Moreover, we propose fixed point algorithms for the special case of Gaussian radial basis function (RBF) kernels. We demonstrate the power of the proposed methods in face and facial expression recognition applications. PMID- 20028627 TI - Multichannel blind separation and deconvolution of images for document analysis. AB - In this paper, we apply Bayesian blind source separation (BSS) from noisy convolutive mixtures to jointly separate and restore source images degraded through unknown blur operators, and then linearly mixed. We found that this problem arises in several image processing applications, among which there are some interesting instances of degraded document analysis. In particular, the convolutive mixture model is proposed for describing multiple views of documents affected by the overlapping of two or more text patterns. We consider two different models, the interchannel model, where the data represent multispectral views of a single-sided document, and the intrachannel model, where the data are given by two sets of multispectral views of the recto and verso side of a document page. In both cases, the aim of the analysis is to recover clean maps of the main foreground text, but also the enhancement and extraction of other document features, such as faint or masked patterns. We adopt Bayesian estimation for all the unknowns and describe the typical local correlation within the individual source images through the use of suitable Gibbs priors, accounting also for well-behaved edges in the images. This a priori information is particularly suitable for the kind of objects depicted in the images treated, i.e., homogeneous texts in homogeneous background, and, as such, is capable to stabilize the ill-posed, inverse problem considered. The method is validated through numerical and real experiments that are representative of various real scenarios. PMID- 20028628 TI - Delay constrained multiplexing of video streams using dual-frame video coding. AB - We consider the multiplexing problem of transmitting multiple video source streams from a server over a shared channel. We use dual-frame video coding with high-quality Long-Term Reference (LTR) frames and propose multiplexing methods to reduce the sum of mean squared error for all the video streams. This paper makes several improvements to dual-frame video coding. A simple motion activity detection algorithm is used to choose the location of LTR frames as well as the number of bits given to such frames. An adaptive buffer-constrained rate-control algorithm is devised to accommodate the extra bits of the high-quality LTR frames. Multiplexing of video streams is studied under the constraint of a video encoder delay buffer. Using H.264/AVC, the results show considerable improvement over baseline schemes such as H.264 rate control when the video streams are encoded individually and over multiplexing methods proposed previously in the literature. The high-quality LTR frames are offset in time among different video streams. This provides the benefits of dual-frame coding with high-quality LTR frames while still fitting under the constraint of an output delay buffer. PMID- 20028629 TI - Video segmentation based on motion coherence of particles in a video sequence. AB - This work describes an approach for object-oriented video segmentation based on motion coherence. Using a tracking process based on adaptively sampled points (namely, particles), 2-D motion patterns are identified with an ensemble clustering approach. Particles are clustered to obtain a pixel-wise segmentation in space and time domains. The segmentation result is mapped to an image spatio temporal feature space. Thus, the different constituent parts of the scene that move coherently along the video sequence are mapped to volumes in this spatio temporal space. These volumes make the redundancy in the temporal sense more explicit, leading to potential gains in video coding applications. The proposed solution is robust and more generic than similar approaches for 2-D video segmentation found in the literature. In order to illustrate the potential advantages of using the proposed motion segmentation approach in video coding applications, the PSNR of the temporal predictions and the entropies of prediction errors obtained in our experiments are presented, and compared with other methods. Our experiments with real and synthetic sequences suggest that our method also could be used in other image processing and computer vision tasks, besides video coding, such as video information retrieval and video understanding. PMID- 20028630 TI - Competitive equilibrium bitrate allocation for multiple video streams. AB - We consider the problem of simultaneous bitrate allocation for multiple video streams. Current methods for multiplexing video streams often rely on identifying the relative complexity of the video streams to improve the combined overall quality. In such methods, not all the videos benefit from the multiplexing process. Typically, the quality of high motion videos is improved at the expense of a reduction in the quality of low motion videos. In our approach, we use a competitive equilibrium allocation of bitrate to improve the quality of all the video streams by finding trades between videos across time. A central controller collects rate-distortion information from each video user and makes a joint bitrate allocation decision. Each user encodes and transmits his video at the allocated bitrate through a shared channel. The proposed method uses information about not only the differing complexity of the video streams at every moment but also the differing complexity of each stream over time. Using the competitive equilibrium bitrate allocation approach for multiple video streams, simulation results show that all the video streams perform better or at least as well as with individual encoding. The results of this research will be useful both for ad hoc networks that employ a cluster head model and for cellular architectures. PMID- 20028631 TI - Contourlet-based image watermarking using optimum detector in a noisy environment. AB - In this paper, an improved multiplicative image watermarking system is presented. Since human visual system is less sensitive to the image edges, watermarking is applied in the contourlet domain, which represents image edges sparsely. In the presented scheme, watermark data is embedded in directional subband with the highest energy. By modeling the contourlet coefficients with General Gaussian Distribution (GGD), the distribution of watermarked noisy coefficients is analytically calculated. The tradeoff between the transparency and robustness of the watermark data is solved in a novel fashion. At the receiver, based on the Maximum Likelihood (ML) decision rule, an optimal detector by the aid of channel side information is proposed. In the next step, a blind extension of the suggested algorithm is presented using the patchwork idea. Experimental results confirm the superiority of the proposed method against common attacks, such as Additive White Gaussian Noise (AWGN), JPEG compression, and rotation attacks, in comparison with the recently proposed techniques. PMID- 20028632 TI - Efficient compression of encrypted grayscale images. AB - Lossless compression of encrypted sources can be achieved through Slepian-Wolf coding. For encrypted real-world sources, such as images, the key to improve the compression efficiency is how the source dependency is exploited. Approaches in the literature that make use of Markov properties in the Slepian-Wolf decoder do not work well for grayscale images. In this correspondence, we propose a resolution progressive compression scheme which compresses an encrypted image progressively in resolution, such that the decoder can observe a low-resolution version of the image, study local statistics based on it, and use the statistics to decode the next resolution level. Good performance is observed both theoretically and experimentally. PMID- 20028633 TI - Coarray synthesis based on polynomial decomposition. AB - Synthetic aperture (SA) techniques have been frequently used to reduce the volume and complexity of the imaging systems. A useful tool for designing synthetic aperture configurations is the coarray. This is the virtual aperture that produces in one way the same beam pattern as the SA system in emission and reception. In this correspondence, we propose a new algorithm, based on the polynomial decomposition, that allows to obtain any wanted coarray on a linear array using whatever synthetic aperture configuration. With this fast and simple algorithm, the desired coarray is decomposed into a set of sub-apertures, whose length is determined by the requirements and resources of the system. The result is the set of weights that have to be applied on the sub-apertures to get the desired coarray, and consequently, the wanted beam pattern. PMID- 20028634 TI - Misalignment-robust face recognition. AB - Subspace learning techniques for face recognition have been widely studied in the past three decades. In this paper, we study the problem of general subspace-based face recognition under the scenarios with spatial misalignments and/or image occlusions. For a given subspace derived from training data in a supervised, unsupervised, or semi-supervised manner, the embedding of a new datum and its underlying spatial misalignment parameters are simultaneously inferred by solving a constrained l1 norm optimization problem, which minimizes the l1 error between the misalignment-amended image and the image reconstructed from the given subspace along with its principal complementary subspace. A byproduct of this formulation is the capability to detect the underlying image occlusions. Extensive experiments on spatial misalignment estimation, image occlusion detection, and face recognition with spatial misalignments and/or image occlusions all validate the effectiveness of our proposed general formulation for misalignment-robust face recognition. PMID- 20028635 TI - The lifting scheme for wavelet bi-frames: theory, structure, and algorithm. AB - In this paper, we present the lifting scheme of wavelet bi-frames along with theory analysis, structure, and algorithm. We show how any wavelet bi-frame can be decomposed into a finite sequence of simple filtering steps. This decomposition corresponds to a factorization of a polyphase matrix of a wavelet bi-frame. Based on this concept, we present a new idea for constructing wavelet bi-frames. For the construction of symmetric bi-frames, we use generalized Bernstein basis functions, which enable us to design symmetric prediction and update filters. The construction allows more efficient implementation and provides tools for custom design of wavelet bi-frames. By combining the different designed filters for the prediction and update steps, we can devise practically unlimited forms of wavelet bi-frames. Moreover, we present an algorithm of increasing the number of vanishing moments of bi-framelets to arbitrary order via the presented lifting scheme, which adopts an iterative algorithm and ensures the shortest lifting scheme. Several construction examples are given to illustrate the results. PMID- 20028636 TI - Simultaneous object classification and segmentation with high-order multiple shape models. AB - Shape models (SMs), capturing the common features of a set of training shapes, represent a new incoming object based on its projection onto the corresponding model. Given a set of learned SMs representing different objects classes, and an image with a new shape, this work introduces a joint classification-segmentation framework with a twofold goal. First, to automatically select the SM that best represents the object, and second, to accurately segment the image taking into account both the image information and the features and variations learned from the online selected model. A new energy functional is introduced that simultaneously accomplishes both goals. Model selection is performed based on a shape similarity measure, online determining which model to use at each iteration of the steepest descent minimization, allowing for model switching and adaptation to the data. High-order SMs are used in order to deal with very similar object classes and natural variability within them. Position and transformation invariance is included as part of the modeling as well. The presentation of the framework is complemented with examples for the difficult task of simultaneously classifying and segmenting closely related shapes, such as stages of human activities, in images with severe occlusions. PMID- 20028637 TI - A stochastic HMM-based forecasting model for fuzzy time series. AB - Recently, fuzzy time series have attracted more academic attention than traditional time series due to their capability of dealing with the uncertainty and vagueness inherent in the data collected. The formulation of fuzzy relations is one of the key issues affecting forecasting results. Most of the present works adopt IF-THEN rules for relationship representation, which leads to higher computational overhead and rule redundancy. Sullivan and Woodall proposed a Markov-based formulation and a forecasting model to reduce computational overhead; however, its applicability is limited to handling one-factor problems. In this paper, we propose a novel forecasting model based on the hidden Markov model by enhancing Sullivan and Woodall's work to allow handling of two-factor forecasting problems. Moreover, in order to make the nature of conjecture and randomness of forecasting more realistic, the Monte Carlo method is adopted to estimate the outcome. To test the effectiveness of the resulting stochastic model, we conduct two experiments and compare the results with those from other models. The first experiment consists of forecasting the daily average temperature and cloud density in Taipei, Taiwan, and the second experiment is based on the Taiwan Weighted Stock Index by forecasting the exchange rate of the New Taiwan dollar against the U.S. dollar. In addition to improving forecasting accuracy, the proposed model adheres to the central limit theorem, and thus, the result statistically approximates to the real mean of the target value being forecast. PMID- 20028638 TI - Ankylosing spondylitis and allied diseases in adults and children. PMID- 20028639 TI - Amniotic fluid cytokines predict pregnancy outcome: myth or reality? PMID- 20028640 TI - Factors affecting discontinuation of alendronate treatment in postmenopausal Japanese women with osteoporosis. AB - BACKGROUND: The main challenge to the long-term treatment of osteoporosis with bisphosphonates has been patient adherence. The purposes of this cohort study were to investigate the 3-year outcome of alendronate treatment in postmenopausal Japanese women with osteoporosis and to identify factors that contributed to the discontinuation of alendronate treatment. METHODS: A total of 72 postmenopausal Japanese women with osteoporosis and aged from 58 to 85 years were treated with alendronate in a 3-year trial. Metacarpal bone mineral density, serum alkaline phosphatase, and urinary cross-linked N-terminal telopeptides of type I collagen were monitored, and factors contributing to the discontinuation of alendronate treatment were determined. RESULTS: Fourteen patients dropped out of the trial. The reasons for dropout were side effects (diarrhea [n = 1], gastric symptoms [n = 9], and inflammation of the mouth [n = 1]) or non-compliance (n = 3). Logistic regression analysis showed that the number of prevalent vertebral fractures was a significant factor affecting the discontinuation of alendronate treatment for the reasons listed above. In 58 patients who continued the 3-year treatment, urinary cross-linked N-terminal telopeptides of type I collagen level was reduced by 44.1% at 3 months and serum alkaline phosphatase level was decreased by 11.6%, 11.8%, and 12.5% at 1, 2, and 3 years, respectively. However, metacarpal bone mineral density did not change significantly. CONCLUSION: Alendronate treatment decreased urinary cross-linked N-terminal telopeptides of type I collagen and serum alkaline phosphatase levels, and maintained metacarpal bone mineral density in postmenopausal Japanese women with osteoporosis. The patients adhered well to alendronate treatment in our clinic. The number of prevalent vertebral fractures was an important factor affecting the discontinuation of alendronate treatment due to side effects and non-compliance. PMID- 20028641 TI - Two-glove-finger-balloon dissection of retroperitoneal space for laparoscopic urology. AB - BACKGROUND: We present our experience in retroperitoneal interventions with 2 catheter mounted glove fingers through 2 access points. METHODS: Patients were placed in the lateral decubitus position, and the retroperitoneum was accessed by blunt dissection through a 10-mm transverse skin incision on the posterior axillary line. Another incision was performed through the anterior axillary line, and the surgeon inserted his finger to dissect the muscle layers in the retroperitoneum to develop an initial space to place the glove finger balloon, which was attached to a 14-F Nelaton catheter. RESULTS: This technique was performed on 32 patients. The mean operation time was 57.4 +/- 26.7 minutes. A satisfactory retroperitoneal space for the operation was provided and both balloons inflated to 500 mL. No complications were observed. CONCLUSION: Retroperitoneal laparoscopy using 2 balloons is a safe, cheap, effective minimally invasive procedure, and we believe that the technique described above both decreases both the operation time and cost. PMID- 20028642 TI - Clinical and molecular epidemiology of infective endocarditis in intravenous drug users. AB - BACKGROUND: Infective endocarditis (IE) in intravenous drug users has been increasing in incidence. The major pathogen used to be methicillin-susceptible Staphylococcus aureus, but resistant isolates have also been increasing. This study aimed to investigate the clinical characteristics of IE in intravenous drug users and to evaluate the molecular patterns of methicillin-resistant S. aureus (MRSA) that cause IE in these drug users. METHODS: A total of 37 episodes of IE in intravenous drug users hospitalized from 1980 to 2006 at a 1,250-bed teaching hospital in Southern Taiwan were evaluated retrospectively. The genetic relatedness of S. aureus strains was assessed using pulsed-field gel electrophoresis. Polymerase chain reaction was used to detect Panton-Valentine leukocidin (PVL) and staphylococcal gamma-hemolysin (Hlg), and to determine the staphylococcal chromosomal cassette carrying the mecA methicillin-resistant gene (SCCmec) type. RESULTS: The patients had a mean +/- standard deviation age of 31.5 +/- 9.25 years, with a male predominance of 76%. Hepatitis C was present in all patients. Methicillin-susceptible S. aureus accounted for 76% of infections, and the most common clinical symptoms were fever (97%) and embolic phenomenon (68%). There were 4 MRSA isolates, 3 of which were SCCmec type III. PVL and Hlg genes were found in 2 and 3 MRSA isolates, respectively. Eighty percent similarity was found among the MRSA isolates by pulsed-field gel electrophoresis. CONCLUSION: Our results suggest that coinfection with hepatitis C was common in intravenous drug users with IE, and that molecular patterns of MRSA isolates had high similarity. SCCmec type III, which is usually hospital-acquired, could have caused the community-associated MRSA endocarditis in our patients. PMID- 20028643 TI - Initial clinical experience with surgical technique of robot-assisted transperitoneal laparoscopic partial nephrectomy. AB - BACKGROUND: The incidental finding of small renal masses has increased due to widespread use of computed tomography as a diagnostic procedure. Some patients with either exophytic renal masses less than 4 cm and suboptimal renal function, a solitary kidney and bilateral renal tumors, or genetic predisposition to renal tumors are considered candidates for laparoscopic partial nephrectomy (LPN). A technical difficulty of LPN is performing laparoscopic intracorporeal suturing under the pressure of warm ischemia time. Because robotic systems have been shown to provide easier intracorporeal suturing, we hypothesized that robotic-assisted LPN might improve efficacy. METHODS: Eight patients with a mean age of 41 years and mean tumor size of 2.3 cm underwent robot-assisted LPN between September 2006 and December 2008. Tumor excision and intracorporeal suturing under warm ischemia by renal artery clamp were performed entirely using a robotic system. All perioperative data and pathologic results were reviewed retrospectively. RESULTS: The mean operation time was 160 minutes, and the mean estimated blood loss was 165 mL. The mean warm ischemia time was 33 minutes, and mean postoperative hospital stay was 4.3 days. Average preoperative hemoglobin was 13.0 mg/dL and postoperative hemoglobin was 11.8 mg/dL. Average preoperative creatinine was 1.1 ng/mL and postoperative creatinine was 1.28 ng/mL. There was 1 conversion to laparoscopic nephrectomy due to a positive margin on a frozen section after discussion with family about better oncologic control. The resected lesions included renal cell carcinoma in 5 patients, angiomyolipoma in 2, and a renin secreting renal tumor in 1 patient. CONCLUSION: Robot-assisted LPN is feasible and may be a viable alternative to open or LPN in selected patients with small exophytic renal tumors. Compared with standard LPN, the robotic assisted LPN approach with precise renal reconstruction under a safe warm ischemia time is feasible and can be easily adopted by those with experience in robot-assisted surgery. PMID- 20028644 TI - Elevated amniotic fluid stromal cell-derived factor-1alpha (SDF-1alpha) concentration in mid-gestation as a predictor of adverse birth outcomes. AB - BACKGROUND: This study aimed to predict maternal and neonatal outcomes by measuring mid-trimester amniotic fluid stromal cell-derived factor-1alpha (SDF 1alpha) concentration in healthy women. METHODS: Mid-trimester amniotic fluid samples from healthy women with a singleton pregnancy were obtained at the time of genetic amniocenteses. SDF-1alpha concentrations were determined by enzyme linked immunosorbent assay. Maternal and neonatal characteristics were recorded. RESULTS: A total of 210 samples were collected. According to the SDF-1alpha cutoff value established by the receiver operating characteristic curve analysis (< 6.42 vs. > or = 6.42 pg/mL), there was a trend toward higher preterm birth rate, lower birth weight and lower 1-minute and 5-minute Apgar scores when SDF 1alpha levels increased (p < 0.05). The pair comparison between normal and selected pregnancy disorders (gestational diabetes, pre-eclampsia, and abnormal placentation) showed no statistical significance (p > 0.05). Pearson's correlations of SDF-1alpha to gestational age at delivery (r = -0.151) and birth weight (r = -0.194) were significant (p < 0.05). In the multivariate analysis on mid-trimester SDF-1alpha levels, maternal age at sampling (regression coefficient = -0.163) and 1-minute Apgar score (< 7 vs. > or = 7, regression coefficient = 2.028) were both significant (p < 0.05). CONCLUSION: Increased SDF-1alpha levels in mid-trimester amniotic fluid suggest a possible role in predicting pregnant women at risk of adverse neonatal outcomes including higher preterm birth rate, lower birth weight, and lower Apgar scores. PMID- 20028645 TI - A cardiac pseudo-tumor associated with pseudo-infarction electrocardiographic pattern. AB - The electrocardiographic presentation of Q waves is an important diagnostic indication of myocardial infarction and other differential diagnoses. We report a case in which the electrocardiography revealed significant Q waves in V1 to V4. In addition, echocardiography disclosed an encapsulated mass lesion over the interventricular septum. Rather than confirm a cardiac tumor, magnetic resonance imaging disclosed hypertrophy of the myocardium with the ventricular septum showing prominent thickness > 3 cm. Hypertrophic cardiomyopathy was diagnosed. Cardiac pseudo-tumor with a pseudo-infarction electrocardiography pattern can be a rare presentation of hypertrophic cardiomyopathy. PMID- 20028646 TI - Balloon mitral commissurotomy for mitral stenosis after resolution of a large mobile left atrial thrombus by 2-year warfarin treatment. AB - We report on a patient who was referred for percutaneous transvenous mitral commissurotomy for symptomatic mitral stenosis. However, transthoracic and transesophageal echocardiography both revealed a large mobile left atrial thrombus, occupying almost half of the left atrial body. Because the patient strongly declined surgical mitral treatment, he was administered warfarin for 2 years. Complete resolution of left atrial thrombus was confirmed by follow-up transesophageal echocardiography. Successful percutaneous transvenous mitral commissurotomy was undertaken without complications. PMID- 20028647 TI - Spontaneous regression of lumbar herniated disc. AB - Intervertebral disc herniation of the lumbar spine is a common disease presenting with low back pain and involving nerve root radiculopathy. Some neurological symptoms in the majority of patients frequently improve after a period of conservative treatment. This has been regarded as the result of a decrease of pressure exerted from the herniated disc on neighboring neurostructures and a gradual regression of inflammation. Recently, with advances in magnetic resonance imaging, many reports have demonstrated that the herniated disc has the potential for spontaneous regression. Regression coincided with the improvement of associated symptoms. However, the exact regression mechanism remains unclear. Here, we present 2 cases of lumbar intervertebral disc herniation with spontaneous regression. We review the literature and discuss the possible mechanisms, the precipitating factors of spontaneous disc regression and the proper timing of surgical intervention. PMID- 20028648 TI - Pseudoaneurysm of the iliac artery secondary to Aspergillus infection after kidney transplantation. AB - Infectious complications are the top causes of morbidity and mortality in patients who undergo renal transplantation. We report a patient who received a cadaveric renal transplant in Mainland China. One year post-transplantation, the patient had right buttock pain with radiation to the leg. Swelling and tenderness over the right groin was also found. Magnetic resonance imaging revealed a multilobulated cystic lesion, about 8 x 7 cm, at the right iliac fossa and presacral region extending to the posterior aspect of the graft kidney and up to the right psoas muscle. Drainage of the intra-abdominal abscess was performed. The abscess culture showed presence of Aspergillus spp. The patient had received steroids, tacrolimus and mycophenolate mofetil, which could be a risk factor for fungal infection. The cause of Aspergillus infection in our patient remains unclear. It may have been due to immune system insufficiency of the patient rendering the patient prone to infection. Pseudoaneurysm formation of the internal iliac artery following Aspergillus infection after kidney transplantation is rarely reported. Although it is a dilemma, once a severe situation such as pseudoaneurysm with aspergillosis presents, graft removal is suggested. PMID- 20028649 TI - Adynamic ileus and acute colonic pseudo-obstruction occurring after cesarean section in patients with massive peripartum hemorrhage. AB - The clinical histories (including radiographs) of 4 patients who suffered from significant adynamic ileus or acute colonic pseudo-obstruction after cesarean section are presented. The main manifestations were vomiting, severe colicky pain, and abdominal distension. These can occur immediately after or within 2 days of the operation. Based on our experience, the risk factors for the development of adynamic ileus are significant peripartum hemorrhage leading to unstable hemodynamic status, severe constipation, use of meperidine for pain relief, and overt bowel manipulation. Mild enema and metoclopramide seem to be helpful in facilitating its resolution. Here, we examine how to differentiate mechanical bowel obstruction from adynamic ileus and look at how to prevent the occurrence of adynamic ileus while minimizing its severity and shortening its clinical course. PMID- 20028650 TI - [Advances in adults' gliomas biology, imaging and treatment]. AB - A better understanding of gliomas biology is now leading to a combined histo molecular classification of these tumors. In anaplastic gliomas ongoing studies depend on 1p/19q codeletion status and in glioblastomas on MGMT methylation status. Advanced brain tumor imaging elicits a better identification of gliomas evolutive potential of. In low-grade gliomas, the importance of maximal resection and the role of chemotherapy are being increasingly recognized. In anaplastic gliomas, phase III studies have clarified the respective roles of chemotherapy and radiotherapy. In glioblastomas concomitant chemoradiotherapy is the standard. Most targeted therapies, namely anti-EGFR therapies have failed to demonstrate efficacy but anti-angiogenics are promising. The aim of this review is to discuss the main advances in adults' gliomas biology, imaging and treatment. PMID- 20028651 TI - [Lung cancer: an update in 2010]. AB - The year 2009 has lead to new data clearly impacting therapeutic management strategies in NSCLC. Personalized medicine is becoming a reality for patients with EGFR mutation as well as for the recruitment of patients in certain clinical trials (EML4-ALK translocation, KRAS mutation...). Maintenance trials are based on questionable statistical designs but this approach may have an interest in certain subset of patients. Little improvements are being achieved in SCLC and locally advanced NSCLC. PMID- 20028652 TI - IFN-gamma regulates the requirement for IL-17 in proteoglycan-induced arthritis. AB - The contribution of the proinflammatory cytokines IFN-gamma and IL-17 to the pathogenesis of experimental arthritis is controversial. In proteoglycan (PG) induced arthritis (PGIA), severe arthritis is dependent on the production of IFN gamma, whereas IL-17 is dispensable. In collagen-induced arthritis and Ag-induced arthritis, although high levels of IFN-gamma are secreted, disease is exacerbated in IFN-gamma or IFN-gamma receptor-deficient mice due to the ability of IFN-gamma to suppress IL-17 expression. In the current study, we investigated the effect of IFN-gamma on the IL-17 response and its consequences in PGIA. In PG-immunized IFN gamma(-/-) mice, despite reduction in arthritis, the PG-specific CD4(+) T cell IL 17 response was significantly increased. Elevated IL-17 contributed to development of arthritis, as disease in IFN-gamma/IL-17(-/-) was significantly reduced in comparison with either IFN-gamma(-/-) or IL-17(-/-) mice. A contribution of IFN-gamma and IL-17 to the development of arthritis was also identified in T-bet(-/-) mice. PG-specific CD4(+) T cells from T-bet(-/-) mice produced reduced IFN-gamma and elevated concentrations of IL-17. Both IFN-gamma and IL-17 contribute to arthritis, as T-bet(-/-) mice lacking IL-17 (T-bet/IL-17( /-)) were resistant, whereas wild-type, T-bet(-/-), and IL-17(-/-) mice were susceptible to PGIA. T cell proliferation and autoantibody production did not correlate with development of disease; however, expression of cytokines and chemokines in joint tissues demonstrate that IFN-gamma and IL-17 cooperatively contribute to inflammation. These results demonstrate that both IFN-gamma and IL 17 have the potential to induce PGIA, but it is the strength of the IFN-gamma response that regulates the contribution of each of these Th effector cytokines to disease. PMID- 20028653 TI - B cells delay neutrophil migration toward the site of stimulus: tardiness critical for effective bacillus Calmette-Guerin vaccination against tuberculosis infection in mice. AB - Mutations in the btk gene encoding Bruton's tyrosine kinase cause X-linked immune deficiency, with impaired B lymphocyte function as the major phenotype. Earlier, we demonstrated that CBA/N-xid mice, unlike the wild-type CBA mice, were not protected by bacillus Calmette-Guerin (BCG) vaccination against tuberculosis infection. Because IFN-gamma-producing T cells and activated macrophages are key elements of antituberculosis protection, it remained unclear how the mutation predominantly affecting B cell functions interferes with responses along the T cell-macrophage axis. In this study, we show that B cell deficiency leads to an abnormally rapid neutrophil migration toward the site of external stimulus. Using adoptive cell transfers and B cell genetic knockout, we demonstrate a previously unappreciated capacity of B cells to downregulate neutrophil motility. In our system, an advanced capture of BCG by neutrophils instead of macrophages leads to a significant decrease in numbers of IFN-gamma-producing T cells and impairs BCG performance in X-linked immune-deficient mice. The defect is readily compensated for by the in vivo neutrophil depletion. PMID- 20028654 TI - Invariant NKT cells in hyperplastic skin induce a local immune suppressive environment by IFN-gamma production. AB - NKT cells can promote or inhibit adaptive immune responses. Cutaneous immunity is tightly regulated by cooperation between innate and adaptive immune processes, but the role of NKT cells in regulating cutaneous immunity is largely unknown. In this study, we show, in a mouse model, that skin-infiltrating CD1d-restricted NKT cells in HPV16-E7 transgenic hyperplastic skin produce IFN-gamma, which can prevent rejection of HPV16-E7-expressing skin grafts. Suppression of graft rejection is associated with the accumulation of CD1d(hi)-expressing CD11c(+)F4/80(hi) myeloid cells in hyperplastic skin. Blockade of CD1d, removal of NKT cells, or local inhibition of IFN-gamma signaling is sufficient to restore immune-mediated graft rejection. Thus, inhibition of NKT cell recruitment or function may enable effective immunity against tumor and viral Ags expressed in epithelial cells. PMID- 20028655 TI - Stromal interaction molecules 1 and 2 are key regulators of autoreactive T cell activation in murine autoimmune central nervous system inflammation. AB - Calcium (Ca(2+)) signaling in T lymphocytes is essential for a variety of functions, including the regulation of differentiation, gene transcription, and effector functions. A major Ca(2+) entry pathway in nonexcitable cells, including T cells, is store-operated Ca(2+) entry (SOCE), wherein depletion of intracellular Ca(2+) stores upon receptor stimulation causes subsequent influx of extracellular Ca(2+) across the plasma membrane. Stromal interaction molecule (STIM) 1 is the Ca(2+) sensor in the endoplasmic reticulum, which controls this process, whereas the other STIM isoform, STIM2, coregulates SOCE. Although the contribution of STIM molecules and SOCE to T lymphocyte function is well studied in vitro, their significance for immune processes in vivo has remained largely elusive. In this study, we studied T cell function in mice lacking STIM1 or STIM2 in a model of myelin-oligodendrocyte glycoprotein (MOG(35-55))-induced experimental autoimmune encephalomyelitis (EAE). We found that STIM1 deficiency significantly impaired the generation of neuroantigen-specific T cell responses in vivo with reduced Th1/Th17 responses, resulting in complete protection from EAE. Mice lacking STIM2 developed EAE, but the disease course was ameliorated. This was associated with a reduced clinical peak of disease. Deficiency of STIM2 was associated with an overall reduced proliferative capacity of lymphocytes and a reduction of IFN-gamma/IL-17 production by neuroantigen-specific T cells. Neither STIM1 nor STIM2 deficiency altered the phenotype or function of APCs. These findings reveal a crucial role of STIM-dependent pathways for T cell function and activation under autoimmune inflammatory conditions, establishing them as attractive new molecular therapeutic targets for the treatment of inflammatory and autoimmune disorders. PMID- 20028656 TI - The alarmin cytokine, high mobility group box 1, is produced by viable cardiomyocytes and mediates the lipopolysaccharide-induced myocardial dysfunction via a TLR4/phosphatidylinositol 3-kinase gamma pathway. AB - High mobility group box 1 (HMGB1) is an alarmin actively secreted by immune cells and passively released by necrotic nonimmune cells. HMGB1 has been implicated in both cardiac contractile dysfunction and the lethality associated with sepsis/endotoxemia. The aim of the current study was to assess whether viable cardiomyocytes could produce HMGB1 and whether HMGB1 can affect myocardial contractility. LPS was used as a model of sepsis/endotoxemia in mice and isolated cardiac myocytes. LPS increased myocardial expression of HMGB1 in vivo (immunohistochemistry) and production and secretion of HMGB1 by viable cardiac myocytes in vitro (Western). LPS increased the phosphorylation status of PI3Kgamma in cardiac myocytes, an effect not observed in TLR4(-/-) myocytes. Genetic (PI3Kgamma(-/-)) or pharmacologic (AS605240) blockade of PI3Kgamma ameliorated the LPS-induced 1) cardiomyocyte production and secretion of HMGB1 in vitro and 2) HMGB1 expression in the myocardium in vivo. The LPS-induced depression of myocardial contractility was prevented by the HMGB1 antagonist, A box. Genetic (PI3Kgamma(-/-)) or pharmacologic (AS605240) blockade of PI3Kgamma ameliorated the LPS-induced decrease in myocardial contractility. No evidence of inflammatory infiltrate was noted in any of the in vivo studies. The findings of the current study indicate that 1) LPS can induce HMGB1 secretion by viable cardiac myocytes through a TLR4/PI3Kgamma signaling pathway, and 2) HMGB1 plays a role in the LPS-induced myocardial contractile dysfunction. The results of the current study also have broader implications (i.e., that viable parenchymal cells, such as cardiac myocytes, participate in the alarmin response). PMID- 20028657 TI - The discovery of immunological tolerance: now more than just a laboratory solution. PMID- 20028658 TI - 'Actively acquired tolerance' of foreign cells. 1953. PMID- 20028660 TI - Mechanisms and consequences of ebolavirus-induced lymphocyte apoptosis. AB - Ebolavirus (EBOV) is a member of the filovirus family and causes severe hemorrhagic fever, resulting in death in up to 90% of infected humans. EBOV infection induces massive bystander lymphocyte apoptosis; however, neither the cellular apoptotic pathway(s) nor the systemic implications of lymphocyte apoptosis in EBOV infection are known. In this study, we show data suggesting that EBOV-induced lymphocyte apoptosis in vivo occurs via both the death receptor (extrinsic) and mitochondrial (intrinsic) pathways, as both Fas-associated death domain dominant negative transgenic mice and mice overexpressing bcl-2 were resistant to EBOV-induced lymphocyte apoptosis. Surprisingly, inhibiting lymphocyte apoptosis during EBOV infection did not result in improved animal survival. Furthermore, we show for the first time that hepatocyte apoptosis likely occurs in EBOV infection, and that mice lacking the proapoptotic genes Bim and Bid had reduced hepatocyte apoptosis and liver enzyme levels postinfection. Collectively, these data suggest that EBOV induces multiple proapoptotic stimuli and that blocking lymphocyte apoptosis is not sufficient to improve survival in EBOV infection. These data suggest that hepatocyte apoptosis may play a role in the pathogenesis of EBOV infection, whereas lymphocyte apoptosis appears to be nonessential for EBOV disease progression. PMID- 20028659 TI - Proteases in MHC class I presentation and cross-presentation. AB - Cells that have mutated their genes or are virally infected are a potential threat to a host. Consequently, the immune system has evolved mechanisms for CD8 T lymphocytes to identify such cells and eliminate them. The generation of CD8 T cell responses occurs in two phases, both of which critically involve the process of Ag presentation. In the first phase, sentinel cells gather Ags present in tissues and then present them to naive CD8 T cells in ways that stimulate their maturation into effectors. In the second phase, these effector cells seek out and eliminate the pathological cells. The abnormal cells are identified through their presentation of immunogenic Ags that they are producing. The Ag presentation mechanisms used by the sentinel cells can be different from those in other cells. This article will review these mechanisms with a focus in each case on how antigenic peptides are generated for presentation. PMID- 20028661 TI - Prostaglandin E(2) exerts homeostatic regulation of pulmonary vascular remodeling in allergic airway inflammation. AB - Nonselective inhibition of PG synthesis augments inflammation in mouse models of airway disease, but the roles of individual PGs are not completely clarified. To investigate the role of PGE(2) in a mouse model of airway inflammation induced by a natural allergen, we used mice lacking the critical terminal synthetic enzyme, microsomal PGE(2) synthase (mPGES)-1. Mice lacking mPGES-1 (ptges(-/-) mice) and wild-type C57BL/6 controls were challenged intranasally with low doses of an extract derived from the house dust mite Dermatophagoides farinae (Der f). The levels of PGE(2) in the bronchoalveolar lavage fluids of Der f-treated ptges(-/-) mice were approximately 80% lower than the levels in wild-type controls. Der f induced bronchovascular eosinophilia was modestly enhanced in the ptges(-/-) mice. Both Der f-treated strains showed similar increases in serum IgE and IgG1, as well as comparable levels of Th1, Th2, and Th17 cytokine production by Der f stimulated spleen cells. These findings indicated that mPGES-1-derived PGE(2) was not required for allergen sensitization or development of effector T cell responses. Unexpectedly, the numbers of vascular smooth muscle cells and the thickness of intrapulmonary vessels were both markedly increased in the Der f treated ptges(-/-) mice. These vascular changes were suppressed by the administration of the stable PGE(2) analog 16, 16-dimethyl PGE(2), or of selective agonists of the E-prostanoid (EP) 1, EP2, and EP3 receptors, respectively, for PGE(2). Thus, mPGES-1 and its product, PGE(2), protect the pulmonary vasculature from remodeling during allergen-induced pulmonary inflammation, and these effects may be mediated by more than one EP receptor. PMID- 20028663 TI - Millenium Development Goals. PMID- 20028662 TI - Rapid and selective detection of fatty acylated proteins using omega-alkynyl fatty acids and click chemistry. AB - Progress in understanding the biology of protein fatty acylation has been impeded by the lack of rapid direct detection and identification methods. We first report that a synthetic omega-alkynyl-palmitate analog can be readily and specifically incorporated into GAPDH or mitochondrial 3-hydroxyl-3-methylglutaryl-CoA synthase in vitro and reacted with an azido-biotin probe or the fluorogenic probe 3-azido 7-hydroxycoumarin using click chemistry for rapid detection by Western blotting or flat bed fluorescence scanning. The acylated cysteine residues were confirmed by MS. Second, omega-alkynyl-palmitate is preferentially incorporated into transiently expressed H- or N-Ras proteins (but not nonpalmitoylated K-Ras), compared with omega-alkynyl-myristate or omega-alkynyl-stearate, via an alkali sensitive thioester bond. Third, omega-alkynyl-myristate is specifically incorporated into endogenous co- and posttranslationally myristoylated proteins. The competitive inhibitors 2-bromopalmitate and 2-hydroxymyristate prevented incorporation of omega-alkynyl-palmitate and omega-alkynyl-myristate into palmitoylated and myristoylated proteins, respectively. Labeling cells with omega alkynyl-palmitate does not affect membrane association of N-Ras. Furthermore, the palmitoylation of endogenous proteins including H- and N-Ras could be easily detected using omega-alkynyl-palmitate as label in cultured HeLa, Jurkat, and COS 7 cells, and, promisingly, in mice. The omega-alkynyl-myristate and -palmitate analogs used with click chemistry and azido-probes will be invaluable to study protein acylation in vitro, in cells, and in vivo. PMID- 20028664 TI - Adolescents under rocket fire: when are coping resources significant in reducing emotional distress? AB - Stress reactions and coping resources of adolescents in chronic and acute situations evoked by missile fire were examined. Data were gathered during August 2006 (Second Lebanon War) on a sample of 303 Israeli adolescents living in Northern Israel (acute state) and 114 youths from Sderot and the Negev, an area which has been exposed to frequent rocket attacks in the last seven years (chronic state). State anxiety and psychological distress were measured as stress reactions. Sense of coherence, family sense of coherence, sense of community and level of exposure were investigated as potential explanatory factors in reducing emotional distress. The overall magnitude of variance explanation was found to be different at each state: a relatively high amount explained variance of stress reactions was found in the chronic stress situation, but not in the acute state. These data support the value of developing a model that differentiates stress situations with the aim of understanding patterns of significant resources in moderating stress reactions in each state. PMID- 20028665 TI - Methodological consideration of story telling in qualitative research involving indigenous peoples. AB - BACKGROUND: The use of storytelling in qualitative research involving Inuit compliments the oral tradition of Inuit culture. The objective of the research was to explore the use of qualitative methods to gain understanding of the experience of living with diabetes, with the ultimate goal of better formulating health care delivery and health promotion among Inuit. METHODS: In-depth interviews were analyzed and interpreted using thematic analysis, open coding, and structured narrative analysis. Inuit community members acted as partners through all stages of the research. RESULTS: ''Because the more we understand, the more we're gonna do a prevention on it ... What I want is use my, use my diabetes, what I have ... so that it can be used by other people for prevention because they'll have understanding about it'' - an Inuk storyteller speaks to the value of education in health promotion. Key methodological issues found relevant to improving qualitative research with Indigenous Peoples include: (i) participatory research methods, grounded in principals of equity, through all phases of research; (ii) the presentation of narratives rather than only interpretations of narratives; (iii) understanding of culture, language, and place to frame the interpretation of the stories in the context within which storytellers experience living with their diabetes, and (iv) the value of multiple methods of analyses. INTERPRETATION: This article comments on the challenges of conducting rigorous research in a cross-cultural setting and outlines methodologies that can improve qualitative narrative analyses research. The research highlighted experiences of living with diabetes and the ways in which storytellers coped and negotiated social support. PMID- 20028666 TI - Is 'modern culture' bad for our health and well-being? AB - Evidence is accumulating that well-being in high-income societies may be static or in decline. One influential theory argues that this is because 'modern' societies are influenced by values of materialism, individualism and consumerism. Does this intellectual critique resonate with ordinary people? This article reports on interviews with purposefully selected groups in Scotland, where the relevance of the cultural critique was explored. Participants in the study believed that cultural values such as individualized consumerism do exert a damaging influence on well-being. They suggested that such values are given particular power in the context of widespread social change and increasing inequalities. Nevertheless, they also believed that individuals and communities possess the capacity to resist such trends. This article concludes that efforts to achieve material improvement for disadvantaged people may not suffice in redressing deep-seated inequalities, if the contribution of some subtle but pernicious effects of contemporary culture remains neglected. However, the research does suggest that positive responses are also possible. PMID- 20028667 TI - High school health curriculum and health literacy: Canadian student voices. AB - This study explores the relevance of health literacy, and its development through a health curriculum, as a necessary but insufficient component to facilitate healthy living among adolescents through comprehensive school health models. This paper presents qualitative findings from focus groups with students (N = 33) in four schools toward the end of their experience in a health class that focused on topics related to healthy living, healthy relationships, health information and decision-making. Students reported mostly negative experiences citing repetitive course content, routinely delivered by teachers and passively received by students. As well, students described their experiences of using health information sources beyond the classroom, such as the media. The findings suggest that the curriculum, and particularly its implementation, have had limited effect on health literacy: students' abilities to access, understand, communicate and evaluate health information. The paper concludes with recommendations for improving health education. PMID- 20028668 TI - Primary health care: applying the principles within a community-based participatory health research project that began in a Canadian women's prison. AB - OBJECTIVES: the purpose of this research was to determine the feasibility of engaging incarcerated women in community-based participatory research and to identify, by and with the women, the health concerns to be addressed. DESIGN: the integration of primary health care, community-based participatory research, a settings approach to health promotion and transformative action research guided the overall design of this study. PARTICIPANTS: Incarcerated women, correctional centre staff and academic researchers participated collaboratively. Setting. The study was conducted in the main short sentence (two years or less) minimum/medium security women's correctional centre in a Canadian province. RESULTS: In-depth interviews were conducted with 16 incarcerated women; in-depth group interviews were facilitated with 16 correctional centre staff. Twenty-one themes, which emerged from participatory, inductive and content analysis of the data, were presented at a face-to-face meeting attended by 120 incarcerated women, 10 correctional centre staff and 5 academic researchers. Underlying values and principles for the project were identified prior to a discussion of the results. During the course of this meeting, the themes were converged into five major categories: addictions and mental health; HIV, hepatitis and infections; health care in prison; life skills and re-entry into society (including homelessness and housing); and children, family and relationships. Numerous suggestions for health interventions and participatory projects were generated, each relating to one of the five major categories. CONCLUSIONS: this study was unique in that, to our knowledge, no other studies have utilized community-based participatory research methods in which incarcerated women played a role in designing the research questions and tools, collecting the data, analyzing the data, interpreting the data and authoring the publications and presentations. This study demonstrated that it is feasible for incarcerated women to engage in developing and utilizing community-based participatory research methods and that these methods can be grounded in a settings approach to whole prison health promotion. PMID- 20028669 TI - Choosing health, constrained choices. AB - In parallel with the neo-liberal retrenchment of the welfarist state, an increasing emphasis on the responsibility of individuals in managing their own affairs and their well-being has been evident. In the health arena for instance, this was a major theme permeating the UK government's White Paper Choosing Health: Making Healthy Choices Easier (2004), which appealed to an ethos of autonomy and self-actualization through activity and consumption which merited esteem. As a counterpoint to this growing trend of informed responsibilization, constrained choices (constrained agency) provides a useful framework for a judicious balance and sense of proportion between an individual behavioural focus and a focus on societal, systemic, and structural determinants of health and well being. Constrained choices is also a conceptual bridge between responsibilization and population health which could be further developed within an integrative biosocial perspective one might refer to as the social ecology of health and disease. PMID- 20028670 TI - Health promotion and surveillance: the establishment of an IUHPE global working group. AB - Following a series of international meetings on behavioral monitoring and surveillance, in 2007 the Italian Ministry of Health (Ministerio della Salute) and the Institute for Health (Istituto Superiore della Sanita) hosted the 5(th) International Conference on Behavioral Risk Factor Surveillance (BRFS) in Rome. A key focus of the conference was on how current surveillance systems could be applied to the field of health promotion, particularly in building the evidence base for health promotion practice. As a result of these discussions, the World Alliance for Risk Factor Surveillance (WARFS), an IUHPE Global Working Group, was formed to work toward providing knowledge and expertise in surveillance as a tool for advancing health promotion. For those IUHPE members interested in participation, this article provides an overview on the strategic direction of WARFS and the newly formed sub-working groups. PMID- 20028671 TI - A healthy nutrition programme with child 'Cub Scouts'. AB - This commentary describes the development of the Health Nutrition programme carried out by University of Cagliari with 19 Italian scout groups. In total 353 children between the ages of 6 and 10 participated in the programme. The objectives were to develop children's knowledge on the Mediterranean diet through games. The project was developed between January and June of 2006. The activities were carried out by the Cub Scout leaders with the objective to improve the Cub Scouts' knowledge on nutrition. Every week they implemented a different game, which showed the Cub Scouts' increased levels of interest and participation. This experience demonstrated that it is possible to involve children in a context outside of the school to carry out a health promotion programme. PMID- 20028672 TI - The integration of bio-medicine and culturally based alternative medicine: implications for health care providers and patients. AB - Complementary and alternative medicine (CAM) are therapies used along with or in place of bio-medicine. Many forms of CAM originate in culture, referred to as culturally based alternative medicines. Usage of CAM is high with large numbers of patients using CAM for mental health, pain and musculoskeletal problems. Their desire for holistic care may be the impetus for this interest, as alternative care practitioners spend more time analyzing illness symptoms. These factors along with the global migration of immigrants accustomed to traditional medicine but now immersed in biomedical health care systems, has created potential for misunderstanding. Drug interactions for some forms of CAM taken with bio-medicine can occur. Insufficient scientific studies about CAM has reduced acceptance and educational opportunities to learn about CAM are limited. Ideas for policy and research are forming. PMID- 20028673 TI - Creating the document 'Promoting health in schools: from evidence to action'. AB - Schools across the world have been involved in health promotion and health education for nearly a century. Do school based initiatives make any difference to the education and health outcomes of young people? This article describes the process in developing the document Promoting health in schools: from evidence to action. The document was produced primarily for the Education sector. It develops an argument about why schools should be undertaking health related initiatives. It also highlights major findings from the literature about what is possible to achieve in school health and the circumstances under which the gains will occur. Attention is focused both on the evidence from the education sector, e.g. effective schools, learning and teaching approaches, and from the health sector, e.g. a whole of school or Health Promoting School (HPS) approach, as well as identifying outcomes from topic areas such as mental and emotional health, healthy eating and nutrition, physical activity, hygiene, sexual health and relationships, substance use and misuse. PMID- 20028675 TI - [Coordination between national and local initiatives in order to strengthen health promotion]. PMID- 20028677 TI - Resettling refugees and safeguarding their mental health: lessons learned from the Canadian Refugee Resettlement Project. AB - The Ryerson University Refugee Resettlement Project (RRP), a decade-long study of 1348 Southeast Asian refugees who came to Canada between 1979 and 1981, is one of the largest, most comprehensive and longest-lived investigations of refugee resettlement ever carried out. Knowledge gleaned from the RRP about research methodology, about the resettlement experience, about the social costs of resettling refugees, about factors that promote or hinder integration, about risk and protective factors for refugee mental health, and about the refugees' consumption of mental health and social services is summarized in the form of 18 "Lessons." The lessons are offered in order to encourage and stimulate further research, as well to suggest policy and practice innovations that could help make resettlement easier, less costly, more effective, and more humane. PMID- 20028678 TI - Help-seeking for mental health problems in young refugees: a review of the literature with implications for policy, practice, and research. AB - The large and diverse bodies of literature on refugee child and adolescent mental health have not been matched by a commensurate interest in help-seeking. Most help-seeking research has centred on Western and, to a lesser extent, non-refugee ethnic minority adult populations. An emerging child and adolescent help-seeking literature consistently reports widespread underutilization of mental health services by children in the general population. Current research and opinion suggest a similar trend for refugee and other ethnic minority children. While service underutilization appears to be an issue for all children, those from refugee backgrounds may be at increased risk of mental health problems and have greater difficulty accessing mental health care. From a policy and practice perspective, the most important explanation for low uptake of services by refugee families concerns an overall failure of Western mental health systems to accommodate the needs of ethnically diverse populations in general and refugees in particular. In order to effectively plan for the mental health needs of refugee children and adolescents, Western host country governments need a clear understanding of help-seeking behaviour. PMID- 20028679 TI - Acculturation and polysubstance abuse in Arab-American treatment clients. AB - Acculturation to U.S. culture by Latinos and Asian Americans has been associated with increased prevalence of substance abuse. However, little is known about the association between acculturation and substance use among Arab Americans, or more specifically, among Arab-American treatment clients. In 156 Arab-American male treatment clients, we found that higher levels of U.S. acculturation were positively associated with increased prevalence of polysubstance abuse. This first report on a large series of Arab-American clients also found considerable within-group variability. These results can be used to develop treatment plans and work-force training on the importance of U.S. acculturation and variability within Arab Americans. PMID- 20028680 TI - Why do Chinese Canadians not consult mental health services: health status, language or culture? AB - Data from the Canadian Community Health Survey Cycle 1.1 showed that Chinese immigrants to Canada and Chinese individuals born in Canada were less likely than other Canadians to have contacted a health professional for mental health reasons in the previous year in the province of British Columbia. The difference persisted among individuals at moderate to high risk for depressive episode. Both immigrant and Canadian-born Chinese showed similar characteristics of mental health service use. The demographic and health factors that significantly affected their likelihood to consult mental health services included Chinese language ability, restriction in daily activities, frequency of medical consultations, and depression score. Notwithstanding lower levels of mental illness in ethnic Chinese communities, culture emerged as a major factor explaining differences in mental health consultation between Chinese and non Chinese Canadians. PMID- 20028681 TI - Veiled communication: is uncovering necessary for psychiatric assessment? AB - Facial expressions are significant to decipher information during a dialogue and more so in a clinical consultation. Veils (Niqab) worn by Muslim women may pose a clinical dilemma for the psychiatric assessment especially if clinicians are not aware of their religious significance. To investigate whether clinical judgment is affected if full facial expressions are not accessible, we conducted an email survey of psychiatrists and psychologists across the world who frequently work in these situations. Of 25 colleagues contacted 16 responded and 11 of them agreed for their comments to be included in the study. Nine out of 11 believed clinical assessment may be compromised, although respondents were aware of cultural sensitivity around the issue. Two out of 11 however, felt fully able to assess the mental state of a veiled woman. Some professionals reported that they feel unable to assess or treat if the request to take the veil off is declined. This small survey demonstrates the diverse opinions on whether unveiling is necessary for psychiatric assessment. Further qualitative examination of this area is needed to develop wider consensus and guidance to mental health care professionals who may be dealing with these groups. PMID- 20028682 TI - Rational use and rationale for use: psychiatric medication at an Argentine institution for intellectual disability. AB - This article describes obstacles to the rational use of psychiatric drugs in a 1000-bed institution for people with intellectual disability employing the WHO framework for rationality. Quantitative data were collected from 98 charts and qualitative data from 14 individuals in this case study. Eight-three percent of patients were taking psychiatric medications, even though 67% of these had no psychiatric diagnosis. Antipsychotics were the first-order treatment; antidepressants were rarely prescribed. These prescribing patterns are influenced by institutional culture, including an emic construct of retardation as a form of psychosis. Low staff-to-patient ratios, a lack of non-medical treatments, and the hospitalization of patients for social reasons also contribute to medication misuse. PMID- 20028683 TI - Claiming the public soul: representations of Qur'anic healing and psychiatry in the Egyptian print media. AB - Egyptian society is engaged in a culture-wide debate over definitions of abnormality, local constructions of which are rooted in ideas about the body and the soul in relation to society as a whole. This is reflected in the continuing recourse to religious healers or texts, as well as in heated debates over the moral, social, religious and legal status of religious healers, in particular the relatively recent and more orthodox "Qur'anic healers." The present study used a primarily qualitative analysis of Egyptian newspaper articles to explore media portrayals of this debate with a focus on how these contradictory cultural themes are situated and contested. The results show that psychiatric hegemony is reflected in media language that gives primacy to certain discourses over others, but that religious healing and religion in general exert an equal, if not more powerful influence on the form of these media portrayals. Different strategies used to negotiate the tensions between Qur'anic healing and psychiatry by those on both sides of the argument come across in the ways these arguments are portrayed in the media. PMID- 20028684 TI - Commentary. Efficacy of duplex ultrasound surveillance after infrainguinal vein bypass may be enhanced by identification of characteristics predictive of graft stenosis development. PMID- 20028685 TI - Commentary. Midterm outcome after the distal revascularization and interval ligation (DRIL) procedure. PMID- 20028686 TI - Commentary. Endovascular treatment of abdominal aortic aneurysms with the Powerlink Endograft System: influence of placement on the bifurcation and use of a proximal extension on early and late outcomes. PMID- 20028688 TI - Stability of blood biochemistry levels in animal model research: effects of storage condition and time. AB - The purpose of this study was to compare whole blood and plasma in terms of the subsequent accuracy of blood lactate, glucose, lactate dehydrogenase (LDH), creatine phosphokinase (CPK), aspartate aminotransferase (AST), and alanine aminotransferase (ALT) measurement. Blood samples were drawn from 8 male Wistar Kyoto rats. The rats were homologous, weighed 300- 380 g, were housed in the same environment, and were provided with food and water under the same conditions. Blood draws occurred in all rats at same time. The blood specimens were divided into two samples, one to be stored as whole blood (WBS) and one to be stored as plasma (PS). All the blood sample analyses were performed by trained and experienced personnel to ensure that differences in results were due to variation in form in which specimens were stored rather than to technique. The lactate concentration in the WBS group gradually increased over time, intraclass correlation coefficient (ICC) = 0.541, 95% confidence interval (CI; -0.197, 0.893), and was higher than that of the PS group, ICC = 0.897, 95% CI (0.733, 0.976). By contrast, glucose level gradually declined for the WBS group, ICC= 0.367, 95% CI (-2.563, 0.682). Whole blood storage increased measurement variation for lactate, glucose, LDH, and CPK. Plasma storage prolonged the stability of the biochemical components. This study demonstrates the importance of evaluating validity at each stage of developing and testing animal models. PMID- 20028689 TI - Integrated review of cytokines in maternal, cord, and newborn blood: part II-- associations with early infection and increased risk of neurologic damage in preterm infants. AB - A growing body of literature supports the relationship of maternal inflammation with preterm birth and adverse neonatal outcomes, including infection and central nervous system (CNS) dysfunction. Mediators of inflammation, most notably proinflammatory cytokines, have been implicated as having an association with and perhaps playing a causal role in the pathogenesis, leading to adverse neonatal outcomes. Even though the association of cytokines with early adverse neonatal outcomes has been actively pursued as a line of research, there has been little integration of diverse findings across studies. Therefore, the purpose of this systematic review was to appraise and classify empirical evidence from human studies for the association of cytokine levels in blood (serum, plasma, or cells; maternal, cord, or neonatal) with two adverse early outcomes in preterm infants: early infection and increased risk of neurologic damage. The review revealed that the proinflammatory cytokines most frequently linked with sepsis are in the interleukin (IL) 1 family as well as tumor necrosis factor alpha (TNF-alpha) and IL-6. The proinflammatory cytokines most frequently linked to neurologic insult in the reviewed studies were IL-1beta, IL-6, and IL-8. In all cases where IL 1beta was studied, the levels were increased when there was neurologic insult. A better understanding of the relationship of these inflammatory substances with these adverse conditions is needed for the future development of maternal and neonatal biobehavioral nursing research. PMID- 20028690 TI - GonadSAGE: a comprehensive SAGE database for transcript discovery on male embryonic gonad development. AB - SUMMARY: Serial analysis of gene expression (SAGE) provides an alternative, with additional advantages, to microarray gene expression studies. GonadSAGE is the first publicly available web-based SAGE database on male gonad development that covers six male mouse embryonic gonad stages, including E10.5, E11.5, E12.5, E13.5, E15.5 and E17.5. The sequence coverage of each SAGE library is beyond 150K, 'which is the most extensive sequence-based male gonadal transcriptome to date'. An interactive web interface with customizable parameters is provided for analyzing male gonad transcriptome information. Furthermore, the data can be visualized and analyzed with the other genomic features in the UCSC genome browser. It represents an integrated platform that leads to a better understanding of male gonad development, and allows discovery of related novel targets and regulatory pathways. PMID- 20028692 TI - Regulation of alternative splicing of the receptor for advanced glycation endproducts (RAGE) through G-rich cis-elements and heterogenous nuclear ribonucleoprotein H. AB - Receptor for advanced glycation endproducts (RAGE) is a cell-surface receptor. The binding of ligands to membrane-bound RAGE (mRAGE) evokes cellular responses involved in various pathological processes. Previously, we identified a novel soluble form, endogenous secretory RAGE (esRAGE) generated by alternative 5' splice site selection in intron 9 that leads to extension of exon 9 (exon 9B). Because esRAGE works as an antagonistic decoy receptor, the elucidation of regulatory mechanism of the alternative splicing is important to understand RAGE related pathological processes. Here, we identified G-rich cis-elements within exon 9B for regulation of the alternative splicing using a RAGE minigene. Mutagenesis of the G-rich cis-elements caused a drastic increase in the esRAGE/mRAGE ratio in the minigene-transfected cells and in loss of binding of the RNA motif to heterogenous nuclear ribonucleoprotein (hnRNP) H. On the other hand, the artificial introduction of a G-stretch in exon 9B caused a drastic decrease in the esRAGE/mRAGE ratio accompanied by the binding of hnRNP H to the RNA motif. Thus, the G-stretches within exon 9B regulate RAGE alternative splicing via interaction with hnRNP H. The findings should provide a molecular basis for the development of medicines for RAGE-related disorders that could modulate esRAGE/mRAGE ratio. PMID- 20028693 TI - Cardioprotective effects of mineralocorticoid receptor antagonists at reperfusion. AB - AIMS: Pre-treatment with mineralocorticoid receptor (MR) antagonists is reported to reduce myocardial infarct size from ischaemia/reperfusion. Here, we tested whether the MR antagonists potassium canrenoate and eplerenone could protect in the more clinically relevant schedule of administration at the end of ischaemia. METHODS AND RESULTS: In all models, hearts were subjected to 30 min regional ischaemia followed by 120 min (rabbits 4 h) reperfusion. A bolus of canrenoate 5 min prior to reperfusion in open-chest mice decreased infarct size in a dose dependent manner. Maximum protection was seen at 1 mg/kg where infarction was 18% of that in the control (P < 0.001). Ecto-5'-nucleotidase (CD73) as well as adenosine A(2b) receptor knock-out mice could no longer be protected, suggesting a role for adenosine and the A(2b) receptor in the mechanism. A 1 mg/kg bolus of canrenoate prior reperfusion also reduced infarct size in open-chest rabbits. To explore the underlying mechanisms, we studied isolated rat hearts. Eplerenone (10 microM) at the end of ischaemia was similarly protective in the rat heart and the protection was abolished by co-treatment with inhibitors of the adenosine receptor, protein kinase C, PI3-kinase, and ERK. In addition, eplerenone or canrenoate treatment increased phosphorylation of the pro-survival kinases Akt and ERK1/2 at reperfusion in the rat hearts. CONCLUSION: Taken together, MR antagonists when given at the end of ischaemia are highly effective and potent cardioprotective drugs with a signalling similar to that of ischaemic pre conditioning and, hence, could be a very promising candidate for the treatment of acute myocardial infarction in man. PMID- 20028694 TI - Chronic heart rate reduction with ivabradine improves systolic function of the reperfused heart through a dual mechanism involving a direct mechanical effect and a long-term increase in FKBP12/12.6 expression. AB - AIMS: To investigate the adaptations of left ventricular function and calcium handling to chronic heart rate reduction with ivabradine in the reperfused heart. METHODS AND RESULTS: Rabbits underwent 20 min coronary artery occlusion followed by 3 weeks of reperfusion. Throughout reperfusion, rabbits received ivabradine (10 mg/kg/day) or vehicle (control). Ivabradine reduced heart rate by about 20% and improved both ejection fraction (+35%) and systolic displacement (+26%) after 3 weeks of treatment. Interestingly, this was associated with a two-fold increase expression of FKBP12/12.6. There was no difference in the expressions of phospholamban, SERCA2a, calsequestrin, ryanodine, phospho-ryanodine, and Na(2+)/Ca(2+) exchanger in the two groups. Infarct scar and vascular density were similar in both groups. Administration of a single intravenous bolus of ivabradine (1 mg/kg) in control rabbits at 3 weeks of reperfusion also significantly improved acutely ejection fraction and systolic displacement. CONCLUSION: Chronic heart rate reduction protects the myocardium against ventricular dysfunction induced by myocardial ischaemia followed by 3 weeks of reperfusion. Beyond pure heart rate reduction, ivabradine improves global and regional systolic function of the reperfused heart through a dual mechanism involving a direct mechanical effect and a long-term adaptation in calcium handling, as supported by the increase in FKBP12/12.6 expression. PMID- 20028695 TI - Lymphofollicular myocarditis: an unknown cause of terminal heart failure. PMID- 20028696 TI - Left bundle branch block and mortality in patients with acute heart failure syndrome: a substudy of the EFICA cohort. AB - AIMS: In patients with chronic heart failure (CHF), left bundle branch block (LBBB) is associated with an increased risk of cardiovascular mortality. We aimed to investigate the association of LBBB with short- and long-term outcome in patients discharged after a de novo episode of acute heart failure (AHF) or AHF complicating a mild CHF. METHODS AND RESULTS: Patients with no history of New York Heart Association class III and IV CHF, who were admitted for a severe AHF episode and enrolled in the prospective observational EFICA study (n = 403), were included. Left bundle branch block prevalence was 16%. Patients with LBBB had a higher prevalence of dilated cardiomyopathy (23 vs. 10%, P < 0.005), a higher percentage of AHF episodes without identified precipitating factor (15 vs. 2%, P < 0.001), and were less likely to present increased markers of cardiac injury (41 vs. 56%, P = 0.04). The 4-week mortality was 24.8% with no difference between LBBB and no LBBB patients. Left bundle branch block was however an independent predictor of 1-year mortality in the 4-week survivors [hazards ratio (95% confidence interval) = 2.01 (1.12-3.64), P = 0.02]. CONCLUSION: Long-term outcome of patients surviving a severe episode of de novo AHF or AHF complicating a mild CHF is worsened by LBBB. These patients may constitute a subgroup at high risk in whom specific therapeutic solutions should be investigated. PMID- 20028697 TI - The theory of epidemiologic transition: the origins of a citation classic. AB - In 1971 Abdel R. Omran published his classic paper on the theory of epidemiologic transition. By the mid-1990s, it had become something of a citation classic and was understood as a theoretical statement about the shift from infectious to chronic diseases that supposedly accompanies modernization. However, Omran himself was not directly concerned with the rise of chronic disease; his theory was in fact closely tied to efforts to accelerate fertility decline through health-oriented population control programs. This article uses Omran's extensive published writings as well as primary and secondary sources on population and family planning to place Omran's career in context and reinterpret his theory. We find that "epidemiologic transition" was part of a broader effort to reorient American and international health institutions towards the pervasive population control agenda of the 1960s and 1970s. The theory was integral to the WHO's then controversial efforts to align family planning with health services, as well as to Omran's unsuccessful attempt to create a new sub-discipline of "population epidemiology." However, Omran's theory failed to displace demographic transition theory as the guiding framework for population control. It was mostly overlooked until the early 1990s, when it belatedly became associated with the rise of chronic disease. PMID- 20028699 TI - Measurements and evaluation of the risks due to external radiation exposures and to intake of activated elements for operational staff engaged in the maintenance of medical cyclotrons. AB - The aim of this paper is to assess the activation phenomena and to evaluate the risk of external exposure and intake doses for the maintenance staff of two medical cyclotrons. Two self-shielded cyclotrons are currently operating in the facility for the routine production of (11)C and (18)F. Four radiochemistry laboratories are linked to the cyclotrons by means of shielded radioisotope delivery lines. Radiopharmaceuticals are prepared both for the PET Diagnostic Department, where four CT-PET scanners are operating with a mean patient workload of 40 d(-1) and for [(18)F]FDG external distribution, to provide radiopharmaceuticals for other institutions. In spite of the fact that air contamination inside the radiochemistry laboratories during the synthesis represents the largest 'slice of the pie' in the evaluation of annual intake dose, potential contamination due to the activated particulate, generated during cyclotron irradiation by micro-corrosion of targets and other components potentially struck by the proton beam and generated neutrons, should be considered. In this regard, the most plausible long-lived (T(1/2) > 30 d) radioisotopes formed are: (97)Tc, (56)Co, (57)Co, (58)Co, (60)Co, (49)V, (55)Fe, (109)Cd, (65)Zn and (22)Na. The results for the operating personnel survey has revealed only low-level contamination for (65)Zn in one test, together with minor (18)F intake, probably due to the environmental dispersion of the radioisotope during the [(18)F]FDG synthesis. PMID- 20028698 TI - Changes in the pattern of DNA methylation associate with twin discordance in systemic lupus erythematosus. AB - Monozygotic (MZ) twins are partially concordant for most complex diseases, including autoimmune disorders. Whereas phenotypic concordance can be used to study heritability, discordance suggests the role of non-genetic factors. In autoimmune diseases, environmentally driven epigenetic changes are thought to contribute to their etiology. Here we report the first high-throughput and candidate sequence analyses of DNA methylation to investigate discordance for autoimmune disease in twins. We used a cohort of MZ twins discordant for three diseases whose clinical signs often overlap: systemic lupus erythematosus (SLE), rheumatoid arthritis, and dermatomyositis. Only MZ twins discordant for SLE featured widespread changes in the DNA methylation status of a significant number of genes. Gene ontology analysis revealed enrichment in categories associated with immune function. Individual analysis confirmed the existence of DNA methylation and expression changes in genes relevant to SLE pathogenesis. These changes occurred in parallel with a global decrease in the 5-methylcytosine content that was concomitantly accompanied with changes in DNA methylation and expression levels of ribosomal RNA genes, although no changes in repetitive sequences were found. Our findings not only identify potentially relevant DNA methylation markers for the clinical characterization of SLE patients but also support the notion that epigenetic changes may be critical in the clinical manifestations of autoimmune disease. PMID- 20028700 TI - Nerve injury by needle nerve perforation in regional anaesthesia: does size matter? AB - BACKGROUND: In regional anaesthesia, there is a risk of direct nerve injury. The purpose of this study was to determine whether the diameter of the applied needle is associated with the magnitude of nerve injury after needle nerve perforation. METHODS: In five anaesthetized pigs, the brachial plexus were exposed bilaterally. Up to eight nerves underwent needle nerve perforation using a 24 G pencil-point cannula (small diameter) or a 19 G pencil-point needle (large diameter). After 48 h, the nerves were resected during anaesthesia. The specimens were processed for visual examination and the detection of inflammatory cells, myelin damage and intraneural haematoma. The grade of nerve injury was scored ranging from 0 (no injury) to 4 (severe injury). RESULTS: Forty-eight nerves were examined. The applied injury score was significantly lower in the small-diameter group [median (inter-quartile range) 2.0 (2.0-2.0)] compared with the large diameter group [3.5 (3.0-4.0) P<0.01]. Myelin damage and intraneural haematoma occurred predominantly in the large-diameter group. Signs of post-traumatic regional inflammation were comparable among both groups. CONCLUSIONS: The severity of nerve injury after needle nerve perforation was related to the diameter of the applied cannula. However, no such difference exists for regional inflammation. Functional consequences of these findings need to be determined. Currently, small-diameter cannulae may be advisable for peripheral nerve blocks to minimize the risk of nerve injury in the case of nerve perforation. PMID- 20028701 TI - Carvedilol-induced elevation in cytosolic free Ca(2+) level and apoptosis in SIRC corneal epithelial cells. AB - The effect of the cardiovascular drug carvedilol on cytosolic free Ca(2+) concentrations ([Ca( 2+)](i)) and viability was examined in Statens Seruminstitut rabbit cornea (SIRC) corneal epithelial cells. [Ca(2+)](i) and cell viability were measured using the fluorescent dyes fura-2 and 4-[3-[4-lodophenyl]-2-4(4 nitrophenyl)-2H-5-tetrazolio-1,3-benzene disulfonate] (WST-1), respectively. Carvedilol at concentrations between 1 and 30 microM increased [Ca( 2+)](i) in a concentration-dependent manner. The Ca(2+) signal was reduced partly by removing extracellular Ca(2+). Carvedilol induced Mn(2+) quench of fura-2 fluorescence implicating Ca(2+) influx. The Ca(2+) influx was inhibited by suppression of protein kinase C activity. In Ca(2+)-free medium, after pretreatment with 1 microM thapsigargin (an endoplasmic reticulum Ca( 2+) pump inhibitor), carvedilol induced [Ca(2+)](i) rise was reduced; and conversely, carvedilol pretreatment inhibited a major part of thapsigargin-induced [Ca( 2+)](i) rise. Addition of the phospholipase C inhibitor 1-[6-[[17 beta-3-methoxyestra-1,3,5(10)-trien-17 yl]amino] hexyl]-1H-pyrrole-2,5-dione (U73122; 2 microM) did not change carvedilol-induced [Ca(2+)](i) rise. At concentrations between 5 and 70 microM, carvedilol killed cells in a concentration-dependent manner. The cytotoxic effect of 20 microM carvedilol was not reversed by prechelating cytosolic Ca(2+) with BAPTA/AM. Apoptosis was induced by 5-70 microM carvedilol. Collectively, in SIRC corneal epithelial cells, carvedilol-induced [Ca(2+)](i) rises by causing Ca(2+) release from the endoplasmic reticulum in a phospholipase C-independent manner, and Ca( 2+) influx via protein kinase C-regulated Ca(2+) channels. Carvedilol caused cytotoxicity was mediated by Ca(2+)-independent apoptosis in a concentration-dependent manner. PMID- 20028702 TI - Effects of Yangtze River source water on genomic polymorphisms of male mice detected by RAPD. AB - In order to evaluate the environmental health risk of drinking water from Yangtze River source, randomly amplified polymorphic DNA (RAPD) markers were used to detect the effects of the source water on genomic polymorphisms of hepatic cell of male mice (Mus musculus, ICR). After the mice were fed with source water for 90 days, RAPD-polymerase chain reactions (PCRs) were performed on hepatic genomic DNA using 20 arbitrary primers. Totally, 189 loci were generated, including 151 polymorphic loci. On average, one PCR primer produced 5.3, 4.9 and 4.8 bands for each mouse in the control, the groups fed with source water and BaP solution, respectively. Compared with the control, feeding mice with Yangtze River source water caused 33 new loci to appear and 19 to disappear. Statistical analysis of RAPD printfingers revealed that Yangtze River source water exerted a significant influence on the hepatic genomic polymorphisms of male mice. This study suggests that RAPD is a reliable and sensitive method for the environmental health risk of Yangtze River source water. PMID- 20028703 TI - L-Ascorbate protects rat hepatocytes against sodium arsenite--induced cytotoxicity and oxidative damage. AB - Sodium arsenite-exposed hepatocytes of rat showed higher production of nitric oxide (NO) and increased lipid peroxidation (LPO) level vis-a-vis activity of superoxide dismutase (SOD) and catalase (CAT) were significantly lowered. Subsequently, the cell proliferation index (CPI) and cell viability were also reduced. Treatment with L-ascorbate was found effective in normalizing the arsenic-induced alteration of SOD and CAT activity and LPO level in rat hepatocytes. These observations indicated that L-ascorbate also has potent cytoprotective role as it could reduce the NO production and normalize the cell proliferation and viability of hepatocytes. Therefore, the in vitro study suggested that ascorbic acid is helpful to ameliorate the arsenic-induced cytotoxicity and oxidative stress of rat hepatocytes. PMID- 20028704 TI - Adverse effects of lactational exposure to chlorpyrifos in suckling rats. AB - The present study was undertaken to evaluate the oxidative damage, biochemical and histopathological alterations in sucking rats whose mothers were exposed to the insecticide chlorpyrifos (CPF). Dams were administered CPF, via oral route. Doses equalled 0.01 mg kg(-1) body weight (b.wt.; acceptable daily intake, ADI), 1.00 mg kg(-1) b.wt. (no observed adverse effects level, NOAEL) and 1.35 mg kg( 1) b.wt. (1/100 lethal dose [LD(50)]) from postnatal day 1 until day 20 after delivery. At two high doses of CPF, the body weight gain and relative liver and kidney weight of suckling pups were significantly decreased. Exposure of the mothers to CPF caused increase in lipid peroxidation (LPO) and decrease in superoxide dismutase (SOD) and glutathione-s-transferase (GST) in lactating pups. CPF altered the level of the marker parameters related to the liver and kidneys. Consistent histological changes were found in the liver and kidneys of the subjected pups, especially at the higher doses. The results suggested that the transfer of CPF intoxication through the mother's milk has resulted in oxidative stress and biochemical and histopathological alterations in the suckling pups. The data of this study may be considered as a contribution to the problem of lactational transfer of the relatively less persistent OP pesticides, such as CPF. PMID- 20028705 TI - Sleep disturbance and fatigue in mild relapsing remitting multiple sclerosis patients on chronic immunomodulant therapy: an actigraphic study. AB - BACKGROUND: Poor sleep is common in MS and it contributes to fatigue. The beta interferons produce systemic effects which may not adapt and may induce fatigue. OBJECTIVE: To verify whether subjective poor sleep and fatigue during chronic therapy correspond to reduced sleep efficiency obtained by actigraphy at home. METHODS: 42 ambulatory relapsing remitting MS patients with mild disability were monitored for at least 7 nights. Habitual sleep quality and fatigue were assessed with the MOS sleep measure and the Fatigue Severity Scale. Sleep logs provided daily sleep quality assessments during actigraphy at home. Patients were grouped according to their current treatment: no therapy, glatiramer acetate, IFNbeta 3 times a week, and IFNbeta once a week. RESULTS AND CONCLUSION: Sleep efficiency was reduced by an average of 5% in 2/3 of the nights following IFNbeta injections compared to the other nights, and daily sleep ratings correlated with actigraphy. Patients on glatiramer acetate also showed a lower sleep efficiency than patients without therapy. Actigraphy data were only modestly correlated with MOSsm scores, not with fatigue. Long term adaptation of sleep effects of immunomodulant agents is incomplete and needs to be considered in treatment planning and assessment of sleep in MS. PMID- 20028706 TI - A voxel-based morphometry study of disease severity correlates in relapsing-- remitting multiple sclerosis. AB - Previous studies have shown a preferential loss of grey matter in fronto-temporal regions in patients with multiple sclerosis. Studies of correlates of disease severity are more controversial, because some studies have suggested an association between sensorimotor cortex atrophy and Expanded Disability Status Scale score, while others did not find such a correlation. The objective of this study was to assess the correlation of regional loss of grey matter and white matter with indexes of clinical and radiological severity in relapsing-remitting multiple sclerosis, including the Expanded Disability Status Scale and lesion load. Correlations between Expanded Disability Status Scale, lesion load and disease duration were assessed in 128 patients with relapsing-remitting multiple sclerosis (Expanded Disability Status Scale range 1.0-6.0) using optimized voxel based morphometry. Bilateral loss of grey matter in sensorimotor cortices was correlated with Expanded Disability Status Scale, and tissue loss also involved adjacent white matter, extending along pyramidal tracts to the brainstem. Increasing lesion load was correlated with loss of deep grey matter and white matter. No specific region of grey matter or white matter showed a significant correlation with disease duration. These findings support the hypothesis that motor neuron involvement plays a major role in the progression of physical disability. Lesion load accrual affects mainly highly interconnected subcortical structures, while disease duration has a less significant impact on brain atrophy, probably owing to the inter-subject heterogeneity of the clinical course of the disease. PMID- 20028708 TI - Latitude and multiple sclerosis prevalence: vitamin D reduces risk of Epstein Barr virus infection. PMID- 20028707 TI - Phase II study of oral fingolimod (FTY720) in multiple sclerosis: 3-year results. AB - In a 6-month, placebo-controlled trial, oral fingolimod (FTY720) 1.25 or 5.0 mg, once daily, significantly reduced MRI inflammatory activity and annualized relapse rate compared with placebo in patients with relapsing multiple sclerosis (MS). The objectives were to monitor the 36-month, interim efficacy and safety results of the ongoing extension of this study. In the extension (months 7-36), placebo-treated patients were re-randomized to either dose of fingolimod; fingolimod-treated patients continued at the same dose. During months 15-24, all patients receiving fingolimod 5.0 mg switched to 1.25 mg. Of the 250 patients who entered the extension study, 173 (69%) continued to month 36. Most patients were free from gadolinium-enhanced lesions (88-89%) or new T2 lesions (70-78%) at month 36. Patients receiving continuous fingolimod treatment had sustained low annualized relapse rates of 0.20-0.21, and 68-73% remained relapse-free at month 36. Over 36 months, nasopharyngitis (34%), headache (30%), fatigue (19%) and influenza (18%) were the most commonly reported adverse events. Pulmonary function remained stable and blood pressure was stable after an initial increase (3-5 mmHg) during the first 6 months of fingolimod treatment; serious adverse events included infections and skin cancer. The low MRI and clinical disease activity at 6 months were maintained at 36 months with fingolimod, which was generally well tolerated by most patients. The efficacy and safety of oral fingolimod are being further evaluated in a large phase III MS study programme. PMID- 20028709 TI - Tear analysis in clinically isolated syndrome as new multiple sclerosis criterion. AB - In clinically isolated syndrome (CIS), the detection of oligoclonal bands (OCBs) in cerebrospinal fluid (CSF) is critical for space dissemination validation when magnetic resonance imaging (MRI) diagnostic criteria are not fulfilled. However, lumbar puncture for CSF collection is considered relatively invasive. Previous studies have demonstrated applicability of OCB detection in tears to the diagnosis of multiple sclerosis (MS). The objective of the present study was to assess concordance between OCB detection in tears and in CSF. We have prospectively included patients with CIS and compared results of CSF and tear OCB detection by isoelectric focusing (IEF). Tears were collected using a Schirmer strip. We included 82 patients. For 69 of them, samples were analysable. OCBs were detected in CSF for 63.8% and in tears for 42% of patients. All patients with tear OCBs had CSF OCBs. We suggest that tear OCB detection may replace CSF OCB detection as a diagnostic tool in patients with CIS. This would circumvent the practice of invasive lumbar punctures currently used in MS diagnosis. PMID- 20028710 TI - Psychometrics and normative data for the Multiple Sclerosis Functional Composite: replacing the PASAT with the Symbol Digit Modalities Test. AB - The MS Functional Composite (MSFC) is a continuous scale of neurological disability for patients with multiple sclerosis (MS). Cognition is represented by the Paced Auditory Serial Addition Test (PASAT), although the Symbol Digit Modalities Test (SDMT) has been proposed as a promising alternative. MSFC scores were calculated using either the PASAT or the SDMT with the following reference populations: National Multiple Sclerosis Society (NMSS) Task Force, 400 MS patients, and 100 normal controls. A subgroup of 115 patients was followed longitudinally, with a test-retest interval of 2.3 +/- 1.2 years. Pearson correlations were calculated and analyses of variance (ANOVAs) were used to assess relationships among the MSFC components and composite scores, and differences in performance between patients and controls. Longitudinal changes were also assessed. Logistic regression was performed to determine which MSFC scores are most predictive of diagnosis, course, and work disability. All MSFCs had similar test-retest reliability and correlations with other measures including neurological disability, depression, and fatigue. The SDMT showed slightly better validity with respect to predicting diagnosis, course, and work disability, although the amount of variance accounted for was similar for each version of the MSFC. Our data, derived from a large sample of MS patients and normal controls, supports the validity of both PASAT and SDMT versions of the MSFC. Because the SDMT has slightly better predictive validity and has a relatively easier administration procedure, some clinicians and researchers may wish to replace the PASAT with the SDMT in future calculations of the MSFC using the calculation methods provided in this manuscript. PMID- 20028711 TI - Influence of HLA-DRB1 alleles on the susceptibility and resistance to multiple sclerosis in Japanese patients with respect to anti-aquaporin 4 antibody status. AB - BACKGROUND: Epistatic interactions between human leukocyte antigen (HLA)-DRB1 alleles alter multiple sclerosis (MS) risk in Caucasians. Such interactions have never been studied in Asian MS patients. OBJECTIVE: To investigate the influence of HLA-DRB1 alleles, including epistatic interactions at this locus, in Japanese MS patients with and without the anti-aquaporin 4 (AQP4) antibody. METHODS: The HLA-DRB1 locus was genotyped in 108 MS patients and 127 healthy controls. MS patients were further classified into two groups according to anti-AQP4 antibody status (27 positive and 81 negative). RESULTS: HLA-DRB1*09 (adjusted odds ratio (OR) = 0.243, 95% confidence interval (CI) 0.099-0.533) and HLA-DRB1*01 (adjusted OR = 0.327, 95% CI 0.103-0.873) decreased the incidence of anti-AQP4 antibody negative MS. By contrast, HLA-DRB1*12 increased the risk of anti-AQP4 antibody positive MS (adjusted OR = 3.691, 95% CI 1.233-10.565). Individuals with HLA DRB1*09/15 decreased the risk of anti-AQP4 antibody-negative MS (adjusted OR = 0.164, 95% CI 0.026-0.593), while those with HLA-DRB1*12/15 increased the risk of anti-AQP4 antibody-positive MS (adjusted OR = 10.870, 95% CI 2.004-81.752). CONCLUSIONS: The ability of HLA-DRB1*09 to reduce the risk of anti-AQP4 antibody negative MS may arise from an interaction with HLA-DRB1*15. By contrast, HLA DRB1*12 increases susceptibility to anti-AQP4 antibody-positive MS, possibly via an interaction with HLA-DRB1*15. PMID- 20028712 TI - A randomized trial of memantine as treatment for spasticity in multiple sclerosis. AB - We report the results of a single center randomized, double-blind, placebo controlled, parallel group trial of memantine in adults with multiple sclerosis and spasticity conducted over 12 weeks. Eligible MS patients had to have an Ashworth spasticity rating of 2 or higher in at least one lower extremity muscle group. Subjects were randomized to receive either placebo or memantine 10 mg twice a day. The primary outcome measure for efficacy was the change in Ashworth Spasticity Scale Score. Although well tolerated, memantine treatment did not demonstrate efficacy in treatment of spasticity in this 12-week small exploratory study. PMID- 20028713 TI - Multi-scale modelling in computational biomedicine. AB - The inherent complexity of biomedical systems is well recognized; they are multi scale, multi-science systems, bridging a wide range of temporal and spatial scales. This article reviews the currently emerging field of multi-scale modelling in computational biomedicine. Many exciting multi-scale models exist or are under development. However, an underpinning multi-scale modelling methodology seems to be missing. We propose a direction that complements the classic dynamical systems approach and introduce two distinct case studies, transmission of resistance in human immunodeficiency virus spreading and in-stent restenosis in coronary artery disease. PMID- 20028714 TI - Neuroanatomy of hemispatial neglect and its functional components: a study using voxel-based lesion-symptom mapping. AB - Spatial neglect is a perplexing neuropsychological syndrome, in which patients fail to detect (and/or respond to) stimuli located contralaterally to their (most often right) hemispheric lesion. Neglect is characterized by a wide heterogeneity, and a role for multiple components has been suggested, but the exact nature of the critical components remains unclear. Moreover, many different lesion sites have been reported, leading to enduring controversies about the relative contribution of different cortical and/or subcortical brain regions. Here we report a systematic anatomo-functional study of 80 patients with a focal right hemisphere stroke, who were examined by a series of neuropsychological tests assessing different clinical manifestations of neglect. We first performed a statistical factorial analysis of their behavioural performance across all tests, in order to break down neglect symptoms into coherent profiles of co varying deficits. We then examined the neural correlates of these distinct neglect profiles using a statistical voxel-based lesion-symptom mapping method that correlated the anatomical extent of brain damage with the relative severity of deficits along the different profiles in each patient. Our factorial analysis revealed three main factors explaining 82% of the total variance across all neglect tests, which suggested distinct components related to perceptive/visuo spatial, exploratory/visuo-motor, and allocentric/object-centred aspects of spatial neglect. Our anatomical voxel-based lesion-symptom mapping analysis pointed to specific neural correlates for each of these components, including the right inferior parietal lobule for the perceptive/visuo-spatial component, the right dorsolateral prefrontal cortex for the exploratory/visuo-motor component, and deep temporal lobe regions for the allocentric/object-centred component. By contrast, standard anatomical overlap analysis indicated that subcortical damage to paraventricular white matter tracts was associated with severe neglect encompassing several tests. Taken together, our results provide new support to the view that the clinical manifestations of hemispatial neglect might reflect a combination of distinct components affecting different domains of spatial cognition, and that intra-hemispheric disconnection due to white matter lesions might produce severe neglect by impacting on more than one functional domain. PMID- 20028715 TI - Beyond these insights there are further applications and future challenges. PMID- 20028716 TI - Primary aortobronchial fistula-induced hemoptysis. PMID- 20028717 TI - The efficacy of ultramolecular aqueous dilutions on a wheat germination model as a function of heat and aging-time. AB - This study evaluates the effects of temperature and aging on the efficacy of As(2)O(3) at the 45th decimal potency in a wheat germination model, compared against a control and potentized H(2)O 45*. Each treatment-temperature combination was tested on seeds (Triticum aestivum L.) of Pandas variety, using six Petri dishes (33 seeds/dish) per trial, performing eight trials. Seeds were pre-treated by poisoning with 0.1% As(2)O(3) solution to reduce germination, to allow a better evaluation of homeopathic treatment effects. The outcome variable was the number of non-germinated seeds after 96 h. Temperature effect was investigated by heating each treatment in a water bath for 30 min (at 20, 40 or 70 degrees C), or for 5 min (at 100 degrees C), and that of aging by dividing experimental data, collected over a period of nearly five months, into two groups: early and late experiments. Results seem to show that the efficacy of As(2)O(3) 45* is unaltered at 20 and 40 degrees C, increases at 70 degrees C and decreases at 100 degrees C. As regards aging, a notable difference was found between early trials, with no significant efficacy, and late trials, where As(2)O(3) 45* exhibits a repeated significant effect versus control, except at 100 degrees C. A reduction in variability was observed for As(2)O(3) 45* at 20 degrees C versus control, confirming the findings of previous work. The main conclusion suggested by this experiment is that the efficacy of As(2)O(3) 45* on wheat germination may be influenced by heating degree and seems to have an increasing trend as a function of aging. PMID- 20028718 TI - Total flavonoids extracted from xiaobuxin-tang on the hyperactivity of hypothalamic-pituitary-adrenal axis in chronically stressed rats. AB - Our previous studies have demonstrated that the total flavonoids (XBXT-2) isolated from the extract of Xiaobuxin-Tang (XBXT), a traditional Chinese herbal decoction, ameliorated behavioral alterations and hippocampal dysfunctions in chronically stressed rats. Studies over the last decades have suggested that the hyperactivity of hypothalamic-pituitary-adrenal (HPA) axis is one of the most consistent findings in stress-related depression. Herein, we used the same chronic mild stress model of rats as before to further investigate the effect of XBXT-2 on the hyperactivity of HPA axis, including the stress hormones levels and glucocorticoid receptors (GRs) expression. Our ELISA results showed that chronic administration of XBXT-2 (25, 50 mg kg(-1), p.o., 28 days, the effective doses for behavioral responses) significantly decreased serum corticosterone level and its upstream stress hormone adrenocorticotropic hormone (ACTH) level in chronically stressed rats. Furthermore, western blotting result demonstrated XBXT 2 treatment ameliorated stress-induced decrease of GRs expression in hippocampus, an important target involved in the hyperactivity of HPA axis. These results were similar to that of classic antidepressant imipramine treatment (10 mg kg(-1), p.o.). In conclusion, the modulation of HPA axis produced by XBXT-2, including the inhibition of stress hormones levels and up-regulation of hippocampal GRs expression, may be an important mechanism underlying its antidepressant-like effect in chronically stressed rats. PMID- 20028719 TI - Analyzing Cytotoxic and Apoptogenic Properties of Scutellaria litwinowii Root Extract on Cancer Cell Lines. AB - The Scutellaria species (Lamiaceae) is used as a source of flavonoids to treat a variety of diseases in traditional medicine. In spite of many reports about the cytotoxic and antitumor effects of some species of this genus, anticancer researches on one of the Iranian species S. litwinowii have not yet been conducted. The cytotoxic properties of total methanol extract of S. litwinowii and its fractions were investigated on different cancer cell lines including AGS, HeLa, MCF-7, PC12 and NIH 3T3. Meanwhile, the role of apoptosis in this toxicity was explored. The cells were cultured in DMEM medium and incubated with different concentrations of herb plant extracts. Cell viability was quantitated by MTT assay. Apoptotic cells were determined using propidium iodide staining of DNA fragmentation by flow cytometry (sub-G1 peak). Scutellaria litwinowii inhibited the growth of malignant cells in a dose-dependent manner. Among solvent fractions of S. litwinowii, the methylene chloride fraction was found to be more toxic compared to other fractions. The IC(50) values of this fraction against AGS, HeLa, MCF-7 and PC12 cell lines after 24 h were determined, 121.2 +/- 3.1, 40.9 +/- 2.5, 115.9 +/- 3.5 and 64.5 +/- 3.4 MUg/ml, respectively. Scutellaria litwinowii induced a sub-G1 peak in the flow cytometry histogram of treated cells compared to control cells indicating that apoptotic cell death is involved in S. litwinowii toxicity. Scutellaria litwinowii exerts cytotoxic and proapototic effects in a variety of malignant cell lines and could be considered as a potential chemotherapeutic agent in cancer treatment. PMID- 20028720 TI - Exploring referral systems for injured patients in low-income countries: a case study from Cambodia. AB - Injury is a growing public health concern worldwide. Since severe injuries require urgent treatment, involving smooth, timely patient referral between facilities, strengthening of the referral system would reduce injury mortality. Smooth referral consists of identification of severe cases, organization of transportation, communication between facilities and prompt care at the receiving facility. This study examined these components of referral of injured patients in a representative sample of health centres (HCs) and referral hospitals (RHs) in Cambodia. We analysed data from a survey carried out in 80 HCs and 17 RHs by interview or mailed questionnaire from December 2006 to April 2007. Collected information on referral included the presence of referral guidelines for injured patients, distance of referral, commonly used transportation and its cost, communication with receiving facilities, and fast-tracking at receiving facilities. Formal referral systems were not functioning well in some areas (insufficient communication and underutilization of ambulances), and informal systems were frequently involved (patient transfer by taxi or referral by community volunteers, and treatment by traditional healers) but were not fully integrated into the referral network (traditional healers seldom referred patients to public facilities). The referral distance was long for most of the surveyed facilities and transportation costs were high when transferring from remote areas, even by ambulance. This study identified the weaknesses and strengths of the emergency referral system in Cambodia. Streamlining referral mechanisms will require organization of each component of the referral mechanism by strengthening the existing system and mobilizing local resources, which would allow Cambodia to develop an efficient system at reasonable cost, though it may differ from Western models. Guidelines including these components along with training and supervision, and expansion of the system to cover other disease conditions, would strengthen the health care system as a whole in this country. PMID- 20028721 TI - Explaining variations in survival in breast cancer in the Eastern Region of England. PMID- 20028722 TI - Transesophageal endoscopic ultrasound-guided fine-needle aspiration for the mediastinal staging of extrathoracic tumors: a new perspective. AB - BACKGROUND: Several extrathoracic tumors metastasize to the mediastinum. Mediastinoscopy is the standard method to obtain tissue proof of mediastinal spread, but drawbacks are its invasiveness, requirement for general anesthesia and costs. Transesophageal endoscopic ultrasound-guided fine-needle aspiration (EUS-FNA) is indicated in lung cancer staging guidelines as a minimally invasive alternative for surgical staging. The diagnostic values in patients with suspected mediastinal metastases and various (previous) extrathoracic malignancies were assessed. PATIENTS AND METHODS: Consecutive patients with suspected mediastinal metastases (on computed tomography or positron emission tomography) and an (previous) extrathoracic malignancy underwent EUS-FNA. RESULTS: Seventy-five patients with current (n = 14) or previously diagnosed (n = 61) extrathoracic malignancies were evaluated. EUS-FNA detected mediastinal malignancies in 43 patients (57%) [metastases of extrathoracic tumors, n = 36 (48%); second malignancy (lung cancer), n = 7 (9%)]. Mediastinal metastases were found at subsequent surgical staging in seven patients or during follow-up (one patient). In seven patients, an alternative diagnosis was established. Sensitivity, specificity, accuracy and negative predictive value of EUS-FNA for mediastinal staging were 86%, 100%, 91% and 72%, respectively. CONCLUSION: EUS FNA is a minimally invasive mediastinal staging method for patients with extrathoracic malignancies to confirm nodal metastatic spread and therefore may qualify as an alternative for surgical staging. PMID- 20028723 TI - Removal of deaminated cytosines and detection of in vivo methylation in ancient DNA. AB - DNA sequences determined from ancient organisms have high error rates, primarily due to uracil bases created by cytosine deamination. We use synthetic oligonucleotides, as well as DNA extracted from mammoth and Neandertal remains, to show that treatment with uracil-DNA-glycosylase and endonuclease VIII removes uracil residues from ancient DNA and repairs most of the resulting abasic sites, leaving undamaged parts of the DNA fragments intact. Neandertal DNA sequences determined with this protocol have greatly increased accuracy. In addition, our results demonstrate that Neandertal DNA retains in vivo patterns of CpG methylation, potentially allowing future studies of gene inactivation and imprinting in ancient organisms. PMID- 20028724 TI - The Arabidopsis thaliana HAK5 K+ transporter is required for plant growth and K+ acquisition from low K+ solutions under saline conditions. AB - K(+) uptake in the high-affinity range of concentrations and its components have been widely studied. In Arabidposis thaliana, the AtHAK5 transporter and the AtAKT1 channel have been shown to be the main transport proteins involved in this process. Here, we study the role of these two systems under two important stress conditions: low K(+) supply or the presence of salinity. T-DNA insertion lines disrupting AtHAK5 and AtAKT1 are employed for long-term experiments that allow physiological characterization of the mutant lines. We found that AtHAK5 is required for K(+) absorption necessary to sustain plant growth at low K(+) in the absence as well as in the presence of salinity. Salinity greatly reduced AtHAK5 transcript levels and promoted AtAKT1-mediated K(+) efflux, resulting in an important impairment of K(+) nutrition. Although having a limited capacity, AtHAK5 plays a major role for K(+) acquisition from low K(+) concentrations in the presence of salinity. PMID- 20028725 TI - Burden of illness in the first 3 years of life in an Indian slum. AB - The morbidity and mortality in a cohort of 452 children followed up from birth up to 3 years of age, in an urban slum in India, is described. These children were recruited and followed from March 2002 to September 2006. A prospective morbidity survey was established. There were 1162 child-years of follow-up. The average morbidity rate was 11.26 episodes/child-year. Respiratory infections caused 58.3 and diarrheal disease 18.4% of the illnesses. Respiratory illnesses resulted in 48, 67.5 and 50 days of illnesses, and there were 3.6, 1.64 and 1.16 diarrheal episodes per child in the 3 years, respectively. There were five deaths in the cohort in the 3 years of follow-up. Of the 77 drop-outs 44 were contacted for mortality data. The morbidity in the area is high, comparable to other studies. The mortality is low, and is attributed to the facilitated access to care. PMID- 20028726 TI - Fueling inflammation at tumor microenvironment: the role of multiligand/RAGE axis. AB - The receptor for advanced glycation end products (RAGE), firstly described in 1992, is a single-transmembrane and multiligand member of the immunoglobulin protein family. RAGE engagement produces activation of multiple intracellular signaling mechanisms involved in several inflammation-associated clinical entities, such as diabetes, cancer, renal and heart failures, as well as neurodegenerative diseases. Although RAGE expression has been extensively reported in many cancer types, it is now emerging as a relevant element that can continuously fuel an inflammatory milieu at the tumor microenvironment, thus changing our perception of its contribution to cancer biology. In this review, we will discuss the role of multiligand/RAGE axis, particularly at the multicellular cross talk established in the inflammatory tumor microenvironment. A better understanding of its contribution may provide new targets for tumor management and risk assessment. PMID- 20028727 TI - Lack of benefit for prophylactic drugs of tension-type headache in adults: a systematic review. AB - OBJECTIVE: To assess the efficacy and tolerability of prophylactic drugs for chronic tension-type headache (TTH) in adults. METHODS: We searched several databases from inception to August 2009. We selected randomized trials that reported the effects of prophylactic drugs in patients with TTH, with a pain measure (intensity, frequency, duration, improvement or index) as outcome measure. Two authors independently assessed risk of bias and extracted data from the original reports. A data synthesis was carried out according to the type of medication. RESULTS: We included 44 trials (3399 patients), of which 15 (34.1%) were considered to be of low risk of bias. Main types of medications studied were antidepressants, muscle relaxants, benzodiazepines and vasodilator agents. Overall, antidepressants were no more effective than placebo, and there were no significant differences between different types of antidepressants. There was conflicting evidence about the effectiveness of benzodiazepines and vasodilator agents compared with placebo. Furthermore, there was limited evidence that propranolol had negative effects on depression in TTH patients, when compared with placebo or biofeedback. There was no evidence concerning the effectiveness of muscle relaxants alone or 5-HT receptor agonist compared with placebo. CONCLUSIONS: Overall, antidepressants were no more effective on headache intensity or frequency and analgesic use than placebo. Propranolol seemed to have negative effects on depression in TTH patients when compared with placebo or biofeedback. No evidence was found for the use of muscle relaxants alone or 5-HT receptor agonist. PMID- 20028728 TI - Clinical relevance of C-reactive protein in ankylosing spondylitis and evaluation of the NSAIDs/coxibs' treatment effect on C-reactive protein. AB - OBJECTIVE: To evaluate the clinical relevance of CRP in AS and investigate the treatment effect of NSAIDs/coxibs on CRP. METHODS: Data of 851 patients with painful axial AS participating in two randomized controlled trials evaluating coxibs (celecoxib) and NSAIDs (naproxen/ketoprofen) were used. The clinical relevance of CRP in AS was studied: the number of patients with abnormal CRP at baseline was expressed as a percentage, and the correlation among CRP, patient (age and gender) and disease [Bath AS Disease Activity Index (BASDAI), Bath AS Functional Index (BASFI)] characteristics was investigated. The standardized response mean (SRM) over placebo was used to detect the treatment effect of NSAIDs/coxibs on CRP. RESULTS: The 851 enrolled patients (73% of whom were men) had an active disease, as assessed through functional impairment. Mean (s.d.) baseline CRP was 14 (19) (median 7.2) mg/l; 61% of patients had an increased CRP. The CRP levels were correlated positively with male sex, BASDAI and BASFI and negatively with age (multivariate analysis). There was a significant difference in the changes of CRP between placebo and NSAIDs/coxibs group (P = 0.003; SRM over placebo: 0.25). Such a treatment effect was of higher magnitude in the subgroup of patients with increased CRP and usually of lower magnitude than the treatment effect of other outcome variables such as BASDAI and BASFI. CONCLUSIONS: Increased CRP was frequently observed in patients with painful axial AS and was correlated both with activity and functional severity of the disease. The treatment effect of NSAIDs/coxibs was relevant in the subgroup of patients with increased CRP at baseline. PMID- 20028729 TI - Reliability of dermoscopy in the assessment of patients with Raynaud's phenomenon. AB - OBJECTIVE: Few rheumatologists have access to wide field or video capillaroscopy (abnormal capillaries being highly predictive of CTD), and a key question is whether they should, therefore, purchase a dermatoscope. The aim of this study was to estimate the inter- and intra-observer variability of dermoscopy (magnification 10x) among rheumatologists with little or no experience of the technique. Good reliability is a necessary prerequisite for a test to be a valid clinical or research tool. METHODS: Dermoscopy was performed in all 10 nail folds from 16 subjects with a range of capillary normality/abnormality. The 160 nail fold images thus acquired were made into two PowerPoint presentations, each of 80 images with 16 duplicate slides. Each participating rheumatologist graded one of the sets of 96 images, grading scale (0-3): normal, mildly abnormal/'suspicious', definitely abnormal, grossly abnormal capillaries or 'unclassifiable' when capillaries could not be adequately identified. Data from both presentations were pooled for analysis. RESULTS: Twenty-eight rheumatologists participated in the study. For the decision as to whether an image could be classified or not, the inter- and intra-observer kappa-coefficients were 0.59 (95% CI 0.51, 0.67) and 0.63 (95% CI 0.45, 0.74), respectively. Conditional on being able to classify, the intra-class correlation coefficient for inter- and intra-observer reliability was 0.72 (95% CI 0.66, 0.77) and 0.85 (95% CI 0.82, 0.92), respectively. CONCLUSIONS: Inter- and intra-observer reliability were good, suggesting that with little training, dermoscopy is likely to be a useful technique to identify capillary distortions/underlying CTD. PMID- 20028730 TI - Evaluation of three automated enzyme immunoassays for detection of anti-cyclic citrullinated peptide antibodies in qualitative and quantitative aspects. AB - OBJECTIVE: The anti-cyclic citrullinated peptide (anti-CCP) antibody has been increasingly used in the field of rheumatology, and various manufacturers have developed a variety of anti-CCP assays using mainly ELISA techniques. This study evaluated the performance of recently marketed automated chemiluminescence enzyme immunoassays for anti-CCP. METHODS: We investigated four anti-CCP assays (Diastat anti-CCP ELISA assay, Axsym anti-CCP assay on the Axsym system, the Architect anti-CCP assay on the Architect i2000 system and the Elecsys anti-CCP assay on the Cobas e 411 analyzer). Samples from 64 patients with RA and 152 controls, including patients with various autoimmune diseases, were studied. We assessed the clinical sensitivities and specificities, and compared the qualitative and quantitative results of each anti-CCP assay. RESULTS: Using the manufacturers' cut-off, diagnostic sensitivities ranged from 90.6 to 93.8% and the specificities ranged from 85.5 to 86.8%. The areas under the curve were comparable among the different assays, and qualitative agreements ranged from 97.2 to 99.1%. In the quantitative results, all anti-CCP assays were significantly correlated (P < 0.001), but the correlation coefficient ranged from 0.615 to 0.861. Especially, the correlation coefficients between the automated anti-CCP assays were higher than those between the ELISA assay (Diastat) and the automated assays. CONCLUSIONS: The overall diagnostic performance of the automated anti-CCP assays was comparable, and it provides reliable information on antibody levels, making them useful in monitoring disease activity. PMID- 20028731 TI - Stress impairs optimal behavior in a water foraging choice task in rats. AB - Stress is a biologically significant social-environmental factor that plays a pervasive role in influencing human and animal behaviors. While stress effects on various types of memory are well characterized, its effects on other cognitive functions are relatively unknown. Here, we investigated the effects of acute, uncontrollable stress on subsequent decision-making performance in rats, using a computer vision-based water foraging choice task. Experiencing stress significantly impaired the animals' ability to progressively bias (but not maintain) their responses toward the larger reward when transitioning from equal to unequal reward quantities. Temporary inactivation of the amygdala during stress, however, blocked impairing effects on decision making. PMID- 20028732 TI - Object recognition memory and the rodent hippocampus. AB - In rodents, the novel object recognition task (NOR) has become a benchmark task for assessing recognition memory. Yet, despite its widespread use, a consensus has not developed about which brain structures are important for task performance. We assessed both the anterograde and retrograde effects of hippocampal lesions on performance in the NOR task. Rats received 12 5-min exposures to two identical objects and then received either bilateral lesions of the hippocampus or sham surgery 1 d, 4 wk, or 8 wk after the final exposure. On a retention test 2 wk after surgery, the 1-d and 4-wk hippocampal lesion groups exhibited impaired object recognition memory. In contrast, the 8-wk hippocampal lesion group performed similarly to controls, and both groups exhibited a preference for the novel object. These same rats were then given four postoperative tests using unique object pairs and a 3-h delay between the exposure phase and the test phase. Hippocampal lesions produced moderate and reliable memory impairment. The results suggest that the hippocampus is important for object recognition memory. PMID- 20028733 TI - Distinct roles for dorsal CA3 and CA1 in memory for sequential nonspatial events. AB - Previous studies have suggested that dorsal hippocampal areas CA3 and CA1 are both involved in representing sequences of events that compose unique episodes. However, it is uncertain whether the contribution of CA3 is restricted to spatial information, and it is unclear whether CA1 encodes order per se or contributes by an active maintenance of memories of sequential events. Here, we developed a new behavioral task that examines memory for the order of sequential nonspatial events presented as trial-unique odor pairings. When the interval between odors within a studied pair was brief (3 sec), bilateral dorsal CA3 lesions severely disrupted memory for their order, whereas dorsal CA1 lesions did not affect performance. However, when the inter-item interval was extended to 10 sec, CA1 lesions, as well as CA3 lesions, severely disrupted performance. These findings suggest that the role of CA3 in sequence memory is not limited to spatial information, but rather appears to be a fundamental property of CA3 function. In contrast, CA1 becomes involved when memories for events must be held or sequenced over long intervals. Thus, CA3 and CA1 are both involved in memory for sequential nonspatial events that compose unique experiences, and these areas play different roles that are distinguished by the duration of time that must be bridged between key events. PMID- 20028734 TI - A new role for sterol regulatory element binding protein 1 transcription factors in the regulation of muscle mass and muscle cell differentiation. AB - The role of the transcription factors sterol regulatory element binding protein 1a (SREBP-1a) and SREBP-1c in the regulation of cholesterol and fatty acid metabolism has been well studied; however, little is known about their specific function in muscle. In the present study, analysis of recent microarray data from muscle cells overexpressing SREBP1 suggested that they may play a role in the regulation of myogenesis. We then demonstrated that SREBP-1a and -1c inhibit myoblast-to-myotube differentiation and also induce in vivo and in vitro muscle atrophy. Furthermore, we have identified the transcriptional repressors BHLHB2 and BHLHB3 as mediators of these effects of SREBP-1a and -1c in muscle. Both repressors are SREBP-1 target genes, and they affect the expression of numerous genes involved in the myogenic program. Our findings identify a new role for SREBP-1 transcription factors in muscle, thus linking the control of muscle mass to metabolic pathways. PMID- 20028735 TI - The cyclin-dependent kinase inhibitor p21 is a crucial target for histone deacetylase 1 as a regulator of cellular proliferation. AB - Histone deacetylases (HDACs) are chromatin-modifying enzymes that are involved in the regulation of proliferation, differentiation and development. HDAC inhibitors induce cell cycle arrest, differentiation, or apoptosis in tumor cells and are therefore promising antitumor agents. Numerous genes were found to be deregulated upon HDAC inhibitor treatment; however, the relevant target enzymes are still unidentified. HDAC1 is required for mouse development and unrestricted proliferation of embryonic stem cells. We show here that HDAC1 reversibly regulates cellular proliferation and represses the cyclin-dependent kinase inhibitor p21 in embryonic stem cells. Disruption of the p21 gene rescues the proliferation phenotype of HDAC1(-/-) embryonic stem cells but not the embryonic lethality of HDAC1(-/-) mice. In the absence of HDAC1, mouse embryonic fibroblasts scarcely undergo spontaneous immortalization and display increased p21 expression. Chromatin immunoprecipitation assays demonstrate a direct regulation of the p21 gene by HDAC1 in mouse embryonic fibroblasts. Transformation with simian virus 40 large T antigen or ablation of p21 restores normal immortalization of primary HDAC1(-/-) fibroblasts. Our data demonstrate that repression of the p21 gene is crucial for HDAC1-mediated control of proliferation and immortalization. HDAC1 might therefore be one of the relevant targets for HDAC inhibitors as anticancer drugs. PMID- 20028736 TI - Differential roles for DNA polymerases eta, zeta, and REV1 in lesion bypass of intrastrand versus interstrand DNA cross-links. AB - Translesion DNA synthesis (TLS) is a process whereby specialized DNA polymerases are recruited to bypass DNA lesions that would otherwise stall high-fidelity polymerases. We provide evidence that TLS across cisplatin intrastrand cross links is performed by multiple translesion DNA polymerases. First, we determined that PCNA monoubiquitination by RAD18 is necessary for efficient bypass of cisplatin adducts by the TLS polymerases eta (Poleta), REV1, and zeta (Polzeta) based on the observations that depletion of these proteins individually leads to decreased cell survival, cell cycle arrest in S phase, and activation of the DNA damage response. Second, we showed that in addition to PCNA monoubiquitination by RAD18, the Fanconi anemia core complex is also important for recruitment of REV1 to stalled replication forks in cisplatin treated cells. Third, we present evidence that REV1 and Polzeta are uniquely associated with protection against cisplatin and mitomycin C-induced chromosomal aberrations, and both are necessary for the timely resolution of DNA double-strand breaks associated with repair of DNA interstrand cross-links. Together, our findings indicate that REV1 and Polzeta facilitate repair of interstrand cross-links independently of PCNA monoubiquitination and Poleta, whereas RAD18 plus Poleta, REV1, and Polzeta are all necessary for replicative bypass of cisplatin intrastrand DNA cross-links. PMID- 20028737 TI - NAD(P)H quinone-oxydoreductase 1 protects eukaryotic translation initiation factor 4GI from degradation by the proteasome. AB - The eukaryotic translation initiation factor 4GI (eIF4GI) serves as a central adapter in cap-binding complex assembly. Although eIF4GI has been shown to be sensitive to proteasomal degradation, how the eIF4GI steady-state level is controlled remains unknown. Here, we show that eIF4GI exists in a complex with NAD(P)H quinone-oxydoreductase 1 (NQO1) in cell extracts. Treatment of cells with dicumarol (dicoumarol), a pharmacological inhibitor of NQO1 known to preclude NQO1 binding to its protein partners, provokes eIF4GI degradation by the proteasome. Consistently, the eIF4GI steady-state level also diminishes upon the silencing of NQO1 (by transfection with small interfering RNA), while eIF4GI accumulates upon the overexpression of NQO1 (by transfection with cDNA). We further reveal that treatment of cells with dicumarol frees eIF4GI from mRNA translation initiation complexes due to strong activation of its natural competitor, the translational repressor 4E-BP1. As a consequence of cap-binding complex dissociation and eIF4GI degradation, protein synthesis is dramatically inhibited. Finally, we show that the regulation of eIF4GI stability by the proteasome may be prominent under oxidative stress. Our findings assign NQO1 an original role in the regulation of mRNA translation via the control of eIF4GI stability by the proteasome. PMID- 20028738 TI - Recruitment of the ESCRT machinery to a putative seven-transmembrane-domain receptor is mediated by an arrestin-related protein. AB - Mammalian arrestins have a major role in the intracellular trafficking of seven transmembrane (7TM) receptors. The fungal ambient pH signaling pathway involves an arrestin-related protein, PalF/Rim8, and the ESCRT (endosomal sorting complex required for transport) machinery. We found that in Saccharomyces cerevisiae, Rim8 binds to both the putative 7TM pH sensor Rim21 and the ESCRT-I subunit Vps23. We show that an SXP motif in Rim8 mediates binding to the Vps23 ubiquitin E2 variant (UEV) domain and that a monoubiquitinated residue near the SXP motif contributes to this interaction. We present evidence that Rim8 ubiquitination is dependent on the Rsp5 E3 ubiquitin ligase and triggered upon binding of Vps23 UEV to both the SXP motif and ubiquitin, thus suggesting a two-step binding mechanism. We further show that Rim8 coimmunoprecipitates with ESCRT-I subunits Vps23 and Vps28, supporting the idea that binding of Rim8 to Vps23 mediates the association of Rim8 with the ESCRT-I complex. Fluorescence microscopic analyses indicate that overexpressed Rim8 and Vps23 colocalize at cortical punctate structures, providing additional evidence of the interaction between these two proteins. Strikingly, our findings indicate that evolutionary conserved mechanisms control the recruitment of the ESCRT machinery to Pal/Rim proteins in fungi and retroviral Gag proteins in animal cells. PMID- 20028739 TI - Splicing factor Spf30 assists exosome-mediated gene silencing in fission yeast. AB - Heterochromatin assembly in fission yeast relies on the processing of cognate noncoding RNAs by both the RNA interference and the exosome degradation pathways. Recent evidence indicates that splicing factors facilitate the cotranscriptional processing of centromeric transcripts into small interfering RNAs (siRNAs). In contrast, how the exosome contributes to heterochromatin assembly and whether it also relies upon splicing factors were unknown. We provide here evidence that fission yeast Spf30 is a splicing factor involved in the exosome pathway of heterochromatin silencing. Spf30 and Dis3, the main exosome RNase, colocalize at centromeric heterochromatin and euchromatic genes. At the centromeres, Dis3 helps recruiting Spf30, whose deficiency phenocopies the dis3-54 mutant: heterochromatin is impaired, as evidenced by reduced silencing and the accumulation of polyadenylated centromeric transcripts, but the production of siRNAs appears to be unaffected. Consistent with a direct role, Spf30 binds centromeric transcripts and locates at the centromeres in an RNA-dependent manner. We propose that Spf30, bound to nascent centromeric transcripts, perhaps with other splicing factors, assists their processing by the exosome. Splicing factor intercession may thus be a common feature of gene silencing pathways. PMID- 20028741 TI - Immunotherapy for human renal cell carcinoma by adoptive transfer of autologous transforming growth factor beta-insensitive CD8+ T cells. AB - PURPOSE: Transforming growth factor-beta (TGF-beta) is a potent immunosuppressor that has been associated with tumor evasion from the host immune surveillance and, thus, tumor progression. We tested a novel immunotherapy for human renal cell cancer (RCC) using a technique that involves the adoptive transfer of autologous tumor-reactive, TGF-beta-insensitive CD8(+) T cells into human RCC challenged immunodeficient mice to identify its potent antitumor responses. EXPERIMENTAL DESIGN: The present study was conducted using a one-to-one adoptive transfer strategy to treat tumor-bearing severe combined immunodeficient (SCID/beige) mouse. The SCID/beige mice were humanized with peripheral blood mononuclear cells from patients with RCC (Hu-PBMC-SCID) before adoptive transfer. Autologous CD8(+) T cells were expanded ex vivo using autologous patient's dendritic cells pulsed with the tumor lysate and rendered TGF-beta insensitive by dominant-negative TGF-beta type II receptor. In addition, human RCC cell lines were generated using patients' tumor cells injected into SCID/beige mice. RESULTS: Using flow cytometry analysis, we confirmed the expression of the tumor reactive, TGF-beta-insensitive CD8(+) T cells were the effector CD8(+) cells (CD27(-)CDRA(+)). Adoptive transfer of autologous TGF-beta-insensitive CD8(+) T cells into tumor-bearing Hu-PBMC-SCID mice induced robust tumor-specific CTL responses in vitro, were associated with tumor apoptosis, suppressed lung metastasis, and prolonged survival times in vivo. CONCLUSION: The one-to-one adoptive transfer strategy is an ideal in vivo murine model for studying the relationship between TGF-beta and immunosurveillance in RCC in vivo. Furthermore, this technique may offer the promise of a novel therapeutic option for the treatment of human patients with RCC. PMID- 20028740 TI - cAMP analogs and their metabolites enhance TREK-1 mRNA and K+ current expression in adrenocortical cells. AB - bTREK-1 K(+) channels set the resting membrane potential of bovine adrenal zona fasciculata (AZF) cells and function pivotally in the physiology of cortisol secretion. Adrenocorticotropic hormone controls the function and expression of bTREK-1 channels through signaling mechanisms that may involve cAMP and downstream effectors including protein kinase A (PKA) and exchange protein 2 directly activated by cAMP (Epac2). Using patch-clamp and Northern blot analysis, we explored the regulation of bTREK-1 mRNA and K(+) current expression by cAMP analogs and several of their putative metabolites in bovine AZF cells. At concentrations sufficient to activate both PKA and Epac2, 8-bromoadenosine-cAMP enhanced the expression of both bTREK-1 mRNA and K(+) current. N(6) Benzoyladenosine-cAMP, which activates PKA but not Epac, also enhanced the expression of bTREK-1 mRNA and K(+) current measured at times from 24 to 96 h. An Epac-selective cAMP analog, 8-(4-chlorophenylthio)-2'-O-methyl-cAMP (8CPT-2'-OMe cAMP), potently stimulated bTREK-1 mRNA and K(+) current expression, whereas the nonhydrolyzable Epac activator 8-(4-chlorophenylthio)-2'-O-methyl-cAMP, Sp-isomer was ineffective. Metabolites of 8CPT-2'-OMe-cAMP, including 8-(4 chlorophenylthio)-2'-O-methyladenosine-5'-O-monophosphate and 8CPT-2'-OMe adenosine, promoted the expression of bTREK-1 transcripts and ion current with a temporal pattern, potency, and effectiveness resembling that of the parent compound. Likewise, at low concentrations, 8-(4-chlorophenylthio)-cAMP (8CPT cAMP; 30 microM) but not its nonhydrolyzable analog 8-(4-chlorophenylthio)-cAMP, Sp-isomer, enhanced the expression of bTREK-1 mRNA and current. 8CPT-cAMP metabolites, including 8CPT-adenosine and 8CPT-adenine, also increased bTREK-1 expression. These results indicate that cAMP increases the expression of bTREK-1 mRNA and K(+) current through a cAMP-dependent but Epac2-independent mechanism. They further demonstrate that one or more metabolites of 8-(4-chlorophenylthio) cAMP analogs potently stimulate bTREK-1 expression by activation of a novel cAMP independent mechanism. These findings raise significant questions regarding the specificity of 8-(4-chlorophenylthio)-cAMP analogs as cAMP mimetics. PMID- 20028742 TI - Targeting of bone-derived insulin-like growth factor-II by a human neutralizing antibody suppresses the growth of prostate cancer cells in a human bone environment. AB - PURPOSE: Advanced prostate cancer frequently involves the bone, where the insulin like growth factor (IGF)-II is abundant. However, the importance of IGF-II in bone metastasis from prostate cancer is uncertain. The present study was aimed at examining the therapeutic importance of targeting IGF-II in bone metastases from prostate cancer. EXPERIMENTAL DESIGN: We investigated whether inhibiting IGF-II using a human neutralizing antibody (m610) suppresses the growth of prostate cancer cells in a human bone environment. Human MDA PCa 2b prostate cancer cells were inoculated into human adult bone implanted into mammary fat pad of nonobese diabetic/severe combined immunodeficient mice or inoculated into mammary fat pad of the mice without human bone implantation. The mice were treated with m610 or a control antibody (m102.4) once weekly for 4 weeks immediately after inoculation with MDA PCa 2b cells. RESULTS: Histomorphologic examination indicated that m610 treatment significantly decreased the MDA PCa 2b tumor area in the human bone compared with the control. Ki-67 immunostaining revealed that the percentage of proliferating cancer cells in the m610-treated bone tumor sections was significantly lower than that in the control. m610 had no effect on MDA PCa 2b tumor growth in the absence of implanted human bone. m610 prevented the in vitro IGF-II-induced proliferation of MDA PCa 2b cells. CONCLUSIONS: Our results indicate that IGF-II plays an important role in the prostate cancer cell growth in human bone, suggesting that targeting it by neutralizing antibodies offers a new therapeutic strategy for bone metastasis from prostate cancer. PMID- 20028743 TI - Mining tissue microarray data to uncover combinations of biomarker expression patterns that improve intermediate staging and grading of clear cell renal cell cancer. AB - PURPOSE: Tumor stage and nuclear grade are the most important prognostic parameters of clear cell renal cell carcinoma (ccRCC). The progression risk of ccRCC remains difficult to predict particularly for tumors with organ-confined stage and intermediate differentiation grade. Elucidating molecular pathways deregulated in ccRCC may point to novel prognostic parameters that facilitate planning of therapeutic approaches. EXPERIMENTAL DESIGN: Using tissue microarrays, expression patterns of 15 different proteins were evaluated in over 800 ccRCC patients to analyze pathways reported to be physiologically controlled by the tumor suppressors von Hippel-Lindau protein and phosphatase and tensin homologue (PTEN). Tumor staging and grading were improved by performing variable selection using Cox regression and a recursive bootstrap elimination scheme. RESULTS: Patients with pT2 and pT3 tumors that were p27 and CAIX positive had a better outcome than those with all remaining marker combinations. A prolonged survival among patients with intermediate grade (grade 2) correlated with both nuclear p27 and cytoplasmic PTEN expression, as well as with inactive, nonphosphorylated ribosomal protein S6. By applying graphical log-linear modeling for over 700 ccRCC for which the molecular parameters were available, only a weak conditional dependence existed between the expression of p27, PTEN, CAIX, and p S6, suggesting that the dysregulation of several independent pathways are crucial for tumor progression. CONCLUSIONS: The use of recursive bootstrap elimination, as well as graphical log-linear modeling for comprehensive tissue microarray (TMA) data analysis allows the unraveling of complex molecular contexts and may improve predictive evaluations for patients with advanced renal cancer. PMID- 20028744 TI - Inhibitor of differentiation 1 contributes to head and neck squamous cell carcinoma survival via the NF-kappaB/survivin and phosphoinositide 3-kinase/Akt signaling pathways. AB - PURPOSE: A key issue in cancer is apoptosis resistance. However, little is known about the transcription factors that contribute to cellular survival of head and neck squamous cell carcinoma (HNSCC). EXPERIMENTAL DESIGN: Three batches (54, 64, and 38) of HNSCC specimens were used for cellular and molecular analyses to determine the major molecular signaling pathways for cellular survival in HNSCC. Animal models (cell culture and xenografts) were used to verify the importance of apoptosis resistance in HNSCC. RESULTS: Inhibitor of differentiation (Id) family member, Id1, was significantly upregulated in clinical HNSCC specimens and acted to protect keratinocytes from apoptosis. Transfection of HNSCC cells with Id1 in vitro induced the phosphorylation of Akt (p-Akt) via phosphoinositide 3-kinase and increased the expression of survivin via NF-kappaB. Blockage of both pathways by specific inhibitors (LY294002 and IkappaBalphaM, respectively) abrogated Id1 induced cell survival of keratinocytes. In vivo studies showed that increased expression of Id1 allowed nontumorigenic keratinocytes (Rhek-1A) to become tumorigenic in nude mice by increased expression of survival genes such as p-Akt and survivin. More importantly, short interfering RNA for Id1 significantly reduced HNSCC tumor volume of HNSCC in xenograft studies. Analysis of clinical data verified the importance of the Id1 downstream molecule, survivin, in the prognosis of HNSCC patients. CONCLUSIONS: The above data, taken together, suggest that Id1 and its downstream effectors are potential targets for treatment of HNSCC because of their contribution to apoptosis resistance. PMID- 20028745 TI - SPARC is associated with gastric cancer progression and poor survival of patients. AB - PURPOSE: The present study investigated the clinical significance of secreted protein, acidic and rich in cysteine (SPARC), in the development and progression of gastric cancer. EXPERIMENTAL DESIGN: Immunohistochemistry was used to analyze SPARC, integrin beta1, and matrix metalloproteinase (MMP)-2 expression in 436 clinicopathologically characterized gastric cancer cases. RESULTS: SPARC, integrin beta1, and MMP-2 protein levels were upregulated in gastric cancer lesions compared with adjacent noncancerous tissues. SPARC protein was detected in 334 of 436 human gastric cancer cases and was highly expressed in 239 tumors. We also found a positive correlation between expression of SPARC and MMP2, and SPARC and integrin beta1. In stages I, II, and III, the 5-year survival rate of patients with a high expression of SPARC was significantly lower than those in patients with low expression. In stage IV, SPARC expression did not correlate with the 5-year survival rate. Further multivariate analysis suggested that the depth of invasion; lymph node and distant metastasis; tumor-node-metastasis stage; and upregulation of SPARC, MMP-2, and integrin beta1, were independent prognostic indicators for the disease. CONCLUSIONS: Our study provided a basis for the development of a novel biomarker for diagnosis and prognosis of gastric cancer. Expression of SPARC in gastric cancer is significantly associated with lymph node and distant metastasis, high MMP2 expression, high intergrin beta1 expression, and poor prognosis. SPARC, intergrin beta1, and MMP-2 protein could be useful markers to predict tumor progression. PMID- 20028747 TI - The role of the Akt/mTOR pathway in tobacco carcinogen-induced lung tumorigenesis. AB - Lung cancer is the leading cause of cancer-related death in the United States, and 85 to 90% of lung cancer cases are associated with tobacco use. Tobacco components promote lung tumorigenesis through genotoxic effects, as well as through biochemical modulation of signaling pathways such as the Akt/mammalian target of rapamycin (mTOR) pathway that regulates cell proliferation and survival. This review will describe cell surface receptors and other upstream components required for tobacco carcinogen-induced activation of Akt and mTOR. Preclinical studies show that inhibitors of the Akt/mTOR pathway inhibit tumor formation in mouse models of carcinogen-induced lung tumorigenesis. Some of these inhibitors will be highlighted, and their clinical potential for the treatment and prevention of lung cancer will be discussed. PMID- 20028748 TI - Activation of WD repeat and high-mobility group box DNA binding protein 1 in pulmonary and esophageal carcinogenesis. AB - PURPOSE: We attempted to identify novel biomarkers and therapeutic targets for lung and esophageal cancers. EXPERIMENTAL DESIGN: We screened for genes that were overexpressed in a large proportion of lung and esophageal carcinomas using a cDNA microarray representing 27,648 genes or expressed sequence tags. A gene encoding WDHD1, a WD repeat and high-mobility group box DNA binding protein 1, was selected as a candidate. Tumor tissue microarray containing 267 archival non small cell lung cancers and 283 esophageal squamous cell carcinomas (ESCC) was used to investigate the clinicopathologic significance of WDHD1 expression. The role of WDHD1 in cancer cell growth and/or survival was examined by small interfering RNA experiments and cell growth assays. The mechanism of WDHD1 activation through its phosphorylation in cancer cells was examined by immunoprecipitation and kinase assays. RESULTS: Positive WDHD1 immunostaining was associated with a poor prognosis for patients with non-small cell lung cancer (P = 0.0403) as well as ESCC (P = 0.0426). Multivariate analysis indicated it to be an independent prognostic factor for ESCC (P = 0.0104). Suppression of WDHD1 expression with small interfering RNAs effectively suppressed lung and esophageal cancer cell growth. In addition, induction of the exogenous expression of WDHD1 promoted the growth of mammalian cells. AKT1 kinase seemed to phosphorylate and stabilize the WDHD1 protein in cancer cells. CONCLUSIONS: WDHD1 expression is likely to play an important role in lung and esophageal carcinogenesis as a cell cycle regulator and a downstream molecule in the phosphoinositide 3-kinase/AKT pathway, and that WDHD1 is a candidate biomarker and a promising therapeutic target for cancer. PMID- 20028749 TI - Somatic EGFR mutation and gene copy gain as predictive biomarkers for response to tyrosine kinase inhibitors in non-small cell lung cancer. AB - PURPOSE: The aim of this systematic review and meta-analysis was to characterize common EGFR molecular aberrations as potential predictive biomarkers for response to monotherapy with tyrosine kinase inhibitors (TKI) in non-small cell lung cancer (NSCLC). EXPERIMENTAL DESIGN: We systematically identified articles investigating EGFR status [somatic mutational and gene copy aberrations (copy number)] in patients with NSCLC treated with TKIs. Eligible studies had to report complete and partial response rates stratified by EGFR status. We used random effects models for bivariable meta-analysis of sensitivity and specificity; positive and negative likelihood ratios (+LR and -LR, respectively) were also calculated and were considered as secondary end points. RESULTS: Among 222 retrieved articles, 59 were considered eligible for the somatic EGFR mutation meta-analysis (1,020 mutations among 3,101 patients) and 21 were considered eligible for the EGFR gene copy number meta-analysis (542 gene gain among 1,539 patients). EGFR mutations were predictive of response to single-agent TKIs [sensitivity, 0.78; 95% confidence interval (95% CI), 0.74-0.82; specificity, 0.86; 95% CI, 0.82-0.89; +LR, 5.6; -LR, 0.25]. EGFR gene gain was also associated with response to TKIs, albeit with lower sensitivity and specificity. In subgroup analysis, the only recognized trend was for a higher predictive value in Whites compared with East Asians for both mutation and gene copy number. CONCLUSION: This analysis provides empirical evidence that EGFR mutations are sensitive and specific predictors of response to single-agent epidermal growth factor receptor TKIs in advanced NSCLC. The diagnostic performance of mutations seems better than that of EGFR gene gain. PMID- 20028750 TI - Association of epidermal growth factor receptor polymorphism, skin toxicity, and outcome in patients with squamous cell carcinoma of the head and neck receiving cetuximab-docetaxel treatment. AB - PURPOSE: Cetuximab, a monoclonal antibody targeting epidermal growth factor receptor (EGFR), has shown clinical efficacy in squamous cell carcinoma of the head and neck with prolonged progression-free (PFS) and overall survival (OS). In this study, we analyzed whether cetuximab-induced skin rash was correlated with distinct polymorphisms within the EGFR gene known to modulate EGFR expression, ligand binding, or signaling activity. EXPERIMENTAL DESIGN: Fifty-one patients enrolled in a single-arm phase II multicenter study for second-line treatment of recurrent or metastatic squamous cell carcinoma of the head and neck with cetuximab/docetaxel were genotyped for two genetic variations in the EGFR gene, a point substitution G-->A in exon 13 resulting in an amino acid substitution in position 521 (EGFR-R521K) and a CA repeat (CA-SSR) polymorphism in intron 1. Association between genotypes and incidence/grade of skin rash was determined by Fisher's exact test. The predictive value of genotypes for PFS and OS was determined using the log-rank test. RESULTS: Overall, 21 patients (41%) developed skin rash with grade >1 within 6 weeks of treatment. The common EGFR-R521K genotype (G/G) was significantly associated with increased skin toxicity (P = 0.024) and showed a trend toward reduced risk of tumor progression (hazard ratio, 0.55; 95% confidence interval, 0.27-1.08; P = 0.08), whereas no correlation of the EGFR-R521K genotype with OS could be observed (P = 0.20). No significant interaction between CA-SSR polymorphism and skin toxicity, PFS, or OS could be detected. CONCLUSIONS: Our study revealed an influence of the EGFR-R521K genotype on skin toxicity and suggested its relation to clinical activity of cetuximab/docetaxel treatment. PMID- 20028751 TI - c-Jun-NH2-kinase-1 inhibition leads to antitumor activity in ovarian cancer. AB - PURPOSE: To show the functional, clinical, and biological significance of c-Jun NH(2)-kinase (JNK)-1 in ovarian carcinoma. EXPERIMENTAL DESIGN: Analysis of the impact of JNK on 116 epithelial ovarian cancers was conducted. The role of JNK in vitro and in experimental models of ovarian cancer was assessed. We studied the role of N-5-[4-(4-methyl piperazine methyl)-benzoylamido]-2-methylphenyl-4-[3-(4 methyl)-pyridyl]-2-pyrimidine amine (WBZ_4), a novel JNK inhibitor redesigned from imatinib based on targeting wrapping defects, in cell lines and in experimental models of ovarian cancer. RESULTS: We found a significant association of pJNK with progression-free survival in the 116 epithelial ovarian cancers obtained at primary debulking therapy. WBZ_4 led to cell growth inhibition and increased apoptosis in a dose-dependent fashion in four ovarian cancer cell lines. In vivo, whereas imatinib had no effect on tumor growth, WBZ_4 inhibited tumor growth in orthotopic murine models of ovarian cancer. The antitumor effect was further increased in combination with docetaxel. Silencing of JNK-1 with systemically administered siRNA led to significantly reduced tumor weights compared with nonsilencing siRNA controls, indicating that indeed the antitumor effects observed were due to JNK-1 inhibition. CONCLUSIONS: These studies identify JNK-1 as an attractive therapeutic target in ovarian carcinoma and that the redesigned WBZ_4 compound should be considered for further clinical development. PMID- 20028752 TI - Phase 1b study of motesanib, an oral angiogenesis inhibitor, in combination with carboplatin/paclitaxel and/or panitumumab for the treatment of advanced non-small cell lung cancer. AB - PURPOSE: Motesanib is a small-molecule antagonist of vascular endothelial growth factor receptor 1, 2, and 3, platelet-derived growth factor receptor, and Kit. This phase 1b study assessed the safety, maximum tolerated dose (MTD), and pharmacokinetics, and explored the objective response of motesanib plus carboplatin/paclitaxel and/or the fully human anti-epidermal growth factor receptor monoclonal antibody panitumumab in advanced non-small cell lung cancer (NSCLC). EXPERIMENTAL DESIGN: Patients with unresectable NSCLC received sequentially escalating doses of motesanib [50, 125 mg once daily; 75 mg twice daily] orally continuously plus carboplatin/paclitaxel (arm A; first line) or panitumumab (arm B; first and second line) once every 21-day cycle or 125 mg once daily plus carboplatin/paclitaxel and panitumumab (arm C; first line). RESULTS: Forty-five patients received motesanib. Three dose-limiting toxicities occurred: grade 4 pulmonary embolism (n = 1; arm A, 50 mg once daily) and grade 3 deep vein thrombosis (n = 2; arm A, 125 mg once daily; arm C). The MTD was 125 mg once daily. Common motesanib-related adverse events were fatigue (60% of patients), diarrhea (53%), hypertension, (38%), anorexia (27%), and nausea (22%). Three cases of cholecystitis occurred but only in the 75-mg twice-daily schedule, which was subsequently discontinued. At 125 mg once daily, motesanib pharmacokinetics were not markedly changed with carboplatin/paclitaxel coadministration; however, exposure to paclitaxel was moderately increased. The objective response rates were 17%, 0%, and 17% in arms A, B, and C, respectively. CONCLUSIONS: Treatment with motesanib was tolerable when combined with carboplatin/paclitaxel and/or panitumumab, with little effect on motesanib pharmacokinetics at the 125-mg once daily dose level. This dose is being investigated in an ongoing phase 3 study in NSCLC. PMID- 20028753 TI - P-glycoprotein (P-gp/Abcb1), Abcc2, and Abcc3 determine the pharmacokinetics of etoposide. AB - PURPOSE: Despite the extensive use of etoposide for the treatment of different malignant neoplasms, its main pharmacokinetic determinants are not completely defined. We aimed to study the impact of P-glycoprotein (P-gp/ABCB1) and the multidrug resistance proteins ABCC2 (MRP2) and ABCC3 (MRP3) on the pharmacokinetics of etoposide. EXPERIMENTAL DESIGN: Abcb1a/1b(-/-), Abcc2(-/-), Abcc3(-/-), Abcb1a/1b;Abcc2(-/-), and Abcc2;Abcc3(-/-) mice were used to investigate the separate and combined impact of P-gp, Abcc2, and Abcc3 on the in vivo behavior of etoposide. RESULTS: P-gp restricted the oral (re)uptake of unchanged etoposide, and mediated its excretion across the gut wall. In contrast, hepatobiliary excretion was almost entirely dependent on Abcc2. Yet, complete loss of Abcc2 did not result in elevated liver or plasma concentrations of etoposide. Instead, Abcc2(-/-) mice displayed an increased hepatic formation of etoposide glucuronide, which was secreted via Abcc3 from the liver to the blood circulation and eliminated with the urine. Combination Abcc2;Abcc3(-/-) mice had highly increased accumulation of etoposide glucuronide in their livers, whereas both single knockouts did not, indicating that Abcc2 and Abcc3 provide alternative pathways for the hepatic elimination of etoposide glucuronide. CONCLUSIONS: P-gp, ABCC2, and ABCC3 significantly affect the pharmacokinetics of etoposide and/or etoposide glucuronide. Variation in transporter expression or activity may explain the high variation in oral availability of etoposide (25 80%) among cancer patients. However, despite the fact that substantial variations in transporter activity can occur, we believe that cancer patients are often relatively protected from etoposide toxicity due to overlapping functions of these transporters in the elimination of etoposide. PMID- 20028754 TI - Chromosome 8p deletions and 8q gains are associated with tumor progression and poor prognosis in prostate cancer. AB - PURPOSE: Deletions of 8p and gains of 8q belong to the most frequent cytogenetic alterations in prostate cancer. The target genes of these alterations and their biological significance are unknown. EXPERIMENTAL DESIGN: To determine the relationship between chromosome 8 changes, and prostate cancer phenotype and prognosis, a set of 1.954 fully annotated prostate cancers were analyzed in a tissue microarray format by fluorescence in situ hybridization. RESULTS: Both 8p deletions and 8q gains increased in number during different stages of prostate cancer progression. 8p deletions/8q gains were found in 26.1%/4.8% of 1,239 pT(2) cancers, 38.5%/9.8% of 379 pT(3a) cancers, 43.5%/8.9% of 237 pT(3b) cancers, 40.7%/14.8% of 27 pT(4) cancers, 39.1%/34.8% of 23 nodal metastases, 51.9%/33.3% of 27 bone metastases, and 45.5%/59.9% of 22 hormone refractory cancers (P < 0.0001 each). Both 8p deletions and 8q gains were also significantly associated with high Gleason grade and with each other (P < 0.0001 each). In primary tumors, 8p deletions were seen in only 27.3% of 1,882 cancers without 8q gain but in 57.4% of 122 tumors with 8q gain (P < 0.0001). Among cancers treated with radical prostatectomy, 8p deletions (P = 0.003) and 8q gains (P = 0.02) were associated with biochemical tumor recurrence. However, multivariate analysis (including prostate-specific antigen, pT/pN stage, Gleason score, and surgical margin status) did not reveal any statistically independent effect of 8p or 8q alterations on biochemical tumor recurrence. CONCLUSIONS: 8p deletions and 8q gains are relatively rare in early stage prostate cancer but often develop during tumor progression. The prognostic effect does not seem to be strong enough to warrant clinical application. PMID- 20028755 TI - Expression signature developed from a complex series of mouse models accurately predicts human breast cancer survival. AB - PURPOSE: The capability of microarray platform to interrogate thousands of genes has led to the development of molecular diagnostic tools for cancer patients. Although large-scale comparative studies on clinical samples are often limited by the access of human tissues, expression profiling databases of various human cancer types are publicly available for researchers. Given that mouse models have been instrumental to our current understanding of cancer progression, we aimed to test the hypothesis that novel gene signatures possessing predictability in clinical outcome can be derived by coupling genomic analyses in mouse models of cancer with publicly available human cancer data sets. EXPERIMENTAL DESIGN: We established a complex series of syngeneic metastatic animal models using a murine breast cancer cell line. Tumor RNA was hybridized on Affymetrix MouseGenome 430A2.0 GeneChips. With the use of Venn logic, gene signatures that represent metastatic competency were derived and tested against publicly available human breast and lung cancer data sets. RESULTS: Survival analyses showed that the spontaneous metastasis gene signature was significantly associated with metastasis-free and overall survival (P < 0.0005). Consequently, the six-gene model was determined and showed statistical predictability in predicting survival in breast cancer patients. In addition, the model was able to stratify poor from good prognosis for lung cancer patients in most data sets analyzed. CONCLUSIONS: Together, our data support that novel gene signature derived from mouse models of cancer can be used for predicting human cancer outcome. Our approaches set precedence that similar strategies may be used to decipher novel gene signatures for clinical utility. PMID- 20028756 TI - A phase I trial of bortezomib with temozolomide in patients with advanced melanoma: toxicities, antitumor effects, and modulation of therapeutic targets. AB - PURPOSE: Preclinical studies show that bortezomib, a proteasome inhibitor, blocks NF-kappaB activation and, combined with temozolomide, enhances activity against human melanoma xenografts and modulates other critical tumor targets. We initiated a phase I trial of temozolomide plus bortezomib in advanced melanoma. Objectives included defining a maximum tolerated dose for the combination, characterizing biomarker changes reflecting inhibition of both proteasome and NF kappaB activity in blood (if possible tumor), and characterizing antitumor activity. EXPERIMENTAL DESIGN: Cohorts were enrolled onto escalating dose levels of temozolomide (50-75 mg/m(2)) daily, orally, for 6 of 9 weeks and bortezomib (0.75-1.5 mg/m(2)) by i.v. push on days 1, 4, 8, and 11 every 21 days. Peripheral blood mononuclear cells were assayed at specified time points for proteasome inhibition and NF-kappaB biomarker activity. RESULTS: Bortezomib (1.3 mg/m(2)) and temozolomide (75 mg/m(2)) proved to be the maximum tolerated dose. Dose limiting toxicities included neurotoxicity, fatigue, diarrhea, and rash. Nineteen melanoma patients were enrolled onto four dose levels. This melanoma population (17 M1c, 10 elevated lactate dehydrogenase, 12 performance status 1-2) showed only one partial response (8 months) and three with stable disease >or=4 months. A significant reduction in proteasome-specific activity was observed 1 hour after infusion at all bortezomib doses. Changes in NF-kappaB electrophoretic mobility shift assay and circulating chemokines in blood failed to correlate with the schedule/dose of bortezomib, inhibition of proteasome activity, or clinical outcome. CONCLUSIONS: We have defined phase II doses for this schedule of temozolomide with bortezomib. Although proteasome activity was inhibited for a limited time in peripheral blood mononuclear cells, we were unable to show consistent effects on NF-kappaB activation. PMID- 20028757 TI - Aldehyde dehydrogenase 1-positive cancer stem cells mediate metastasis and poor clinical outcome in inflammatory breast cancer. AB - PURPOSE: To examine the role of cancer stem cells (CSC) in mediating metastasis in inflammatory breast cancer (IBC) and the association of these cells with patient outcome in this aggressive type of breast cancer. EXPERIMENTAL DESIGN: CSCs were isolated from SUM149 and MARY-X, an IBC cell line and primary xenograft, by virtue of increased aldehyde dehydrogenase (ALDH) activity as assessed by the ALDEFLUOR assay. Invasion and metastasis of CSC populations were assessed by in vitro and mouse xenograft assays. Expression of ALDH1 was determined on a retrospective series of 109 IBC patients and this was correlated with histoclinical data. All statistical tests were two sided. Log-rank tests using Kaplan-Meier analysis were used to determine the correlation of ALDH1 expression with development of metastasis and patient outcome. RESULTS: Both in vitro and xenograft assays showed that invasion and metastasis in IBC are mediated by a cellular component that displays ALDH activity. Furthermore, expression of ALDH1 in IBC was an independent predictive factor for early metastasis and decreased survival in this patient population. CONCLUSIONS: These results suggest that the metastatic, aggressive behavior of IBC may be mediated by a CSC component that displays ALDH enzymatic activity. ALDH1 expression represents the first independent prognostic marker to predict metastasis and poor patient outcome in IBC. The results illustrate how stem cell research can translate into clinical practice in the IBC field. PMID- 20028758 TI - Decreased blood flow with increased metabolic activity: a novel sign of pancreatic tumor aggressiveness. PMID- 20028759 TI - Single nucleotide polymorphisms of gemcitabine metabolic genes and pancreatic cancer survival and drug toxicity. AB - PURPOSE: To show whether single nucleotide polymorphisms (SNP) of drug metabolic genes were associated with toxicity of 2',2'-difluoro 2'-deoxycytidine (gemcitabine)-based chemoradiotherapy and overall survival (OS) of patients with pancreatic cancer. EXPERIMENTAL DESIGN: We evaluated 17 SNPs of the CDA, dCK, DCTD, RRM1, hCNT1, hCNT2, hCNT3, and hENT1 genes in 154 patients with potentially resectable pancreatic adenocarcinoma who were enrolled in clinical trials at The University of Texas M.D. Anderson Cancer Center (Houston, TX) from February 1999 to January 2006, with follow-up until April 2009. Patients received neoadjuvant concurrent gemcitabine and radiation therapy with or without gemcitabine cisplatin induction therapy. The association of genotypes with toxicity or OS was tested, respectively, by logistic regression and Cox regression analysis. RESULTS: None of the 17 SNPs, individually, had a significant association with OS. A combined genotype effect of CDA A-76C, dCK C-1205T, DCTD T-47C, hCNT3 C 69T, hENT1 T-549C, and hENT1 C913T on OS was observed. Patients carrying 0 to 1 (n = 43), 2 to 3 (n = 77), or 4 to 6 (n = 30) variant alleles had median survival time of 31.5, 21.4, and 17.5 months, respectively. The hazard ratio of dying was 1.71 (95% confidence interval, 1.06-2.76) and 3.16 (95% confidence interval, 1.77 5.63) for patients carrying two to three or four to six at-risk genotypes (P = 0.028 and P < 0.001), respectively, after adjusting for clinical predictors. CDA C111T, dCK C-1205T, dCK A9846G, and hCNT3 A25G, individually and jointly, had a significant association with neutropenia toxicity. CONCLUSIONS: These observations suggest that polymorphic variations of drug metabolic genes were associated with toxicity of gemcitabine-based therapy and OS of patients with resectable pancreatic cancer. PMID- 20028760 TI - Rap1 stabilizes beta-catenin and enhances beta-catenin-dependent transcription and invasion in squamous cell carcinoma of the head and neck. AB - PURPOSE: In head and neck squamous cell carcinoma (HNSCC) cells, Rap1 shuttles between the nucleus and cytoplasm. Prior findings suggested that Rap1 may modulate the beta-catenin-independent Wnt pathway in some settings, but the role of Rap1 in beta-catenin-dependent Wnt signaling remains undefined. EXPERIMENTAL DESIGN AND RESULTS: We observed that beta-catenin bound to active Rap1 in vitro and Rap1 activated beta-catenin/T-cell factor (TCF)-dependent transcription. Immunofluorescence studies showed that ectopic expression of Rap1 increased nuclear translocation of beta-catenin. Overexpression of active Rap1 facilitated an increase in beta-catenin-mediated transcription that was abrogated by dominant negative TCF4. Conversely, small interfering RNA-mediated inhibition of endogenous Rap1 expression inhibited beta-catenin/TCF-mediated transcription as well as invasion of HNSCC. Furthermore, inhibition of Rap1 expression downregulated the expression of matrix metalloproteinase 7, a transcriptional target of beta-catenin/TCF. In HNSCC cells stably transfected with beta-catenin or treated with lithium chloride or Wnt3A to stabilize endogenous beta-catenin, inhibition of Rap1 expression led to decreases in the free pool of beta-catenin. Immunohistochemical studies of tissue from HNSCC patients revealed that increased beta-catenin intensity correlated with higher tumor stage. Furthermore, the prognostic effect of active Rap1 on tumor N stage was found to depend on cytosolic beta-catenin expression (P < 0.013). When beta-catenin is high, higher Rap1GTP intensity is associated with more advanced N stage. CONCLUSIONS: The findings suggest that Rap1 enhances beta-catenin stability and nuclear localization. In addition to indicating that Rap1 has a significant role in regulating beta-catenin and beta-catenin-dependent progression to more advanced N stage lesions, these data highlight Rap1 as a potential therapeutic target in HNSCC. PMID- 20028762 TI - Bevacizumab safety in patients with central nervous system metastases. AB - PURPOSE: Patients with central nervous system (CNS) metastases were excluded from bevacizumab trials following a case of fatal cerebral hemorrhage in a patient with hepatocellular carcinoma in 1997. Safety information for bevacizumab-treated patients with CNS metastases was reviewed to determine whether general exclusion of these patients from bevacizumab treatment is still justified. EXPERIMENTAL DESIGN: A retrospective exploratory analysis was conducted using datasets from 13 randomized controlled phase II/III trials (dataset A), two open-label single-arm safety trials (dataset B), and two prospective studies including patients with treated CNS metastases (dataset C). In datasets A and B, known CNS metastasis was an exclusion criterion; patients with CNS metastasis had unrecognized CNS metastases at study entry or developed them during the trial. All reported cerebral hemorrhage grades in patients with CNS metastases were quantified. RESULTS: In dataset A, occult brain metastases were identified in 187 of 8,443 patients (91 in bevacizumab arms and 96 in non-bevacizumab arms). Three bevacizumab-treated patients (3.3%) developed grade 4 cerebral hemorrhage, whereas one control-arm patient (1.0%) developed grade 5 cerebral hemorrhage. In dataset B, 321 of 4,382 patients had initially occult CNS metastases, in whom two grade 1 and one grade 3 cerebral hemorrhage (0.9%) were reported. In 131 patients with treated CNS metastases in dataset C, one bevacizumab-treated patient (0.8%) developed grade 2 cerebral hemorrhage. CONCLUSIONS: In this selected population, patients with CNS metastases are at similar risk of developing cerebral hemorrhage, independent of bevacizumab therapy. Consequently, such patients with CNS metastases from advanced/metastatic breast cancer, non-small cell lung carcinoma, and renal and colorectal cancer should not be generally excluded from bevacizumab therapy or clinical trials. PMID- 20028761 TI - Immunotherapy of malignant disease with tumor antigen-specific monoclonal antibodies. AB - A few tumor antigen (TA)-specific monoclonal antibodies (mAb) have been approved by the Food and Drug Administration for the treatment of several major malignant diseases and are commercially available. Once in the clinic, mAbs have an average success rate of approximately 30% and are well tolerated. These results have changed the face of cancer therapy, bringing us closer to more specific and more effective biological therapy of cancer. The challenge facing tumor immunologists at present is represented by the identification of the mechanism(s) underlying the patients' differential clinical response to mAb-based immunotherapy. This information is expected to lead to the development of criteria to select patients to be treated with mAb-based immunotherapy. In the past, in vitro and in vivo evidence has shown that TA-specific mAbs can mediate their therapeutic effect by inducing tumor cell apoptosis, inhibiting the targeted antigen function, blocking tumor cell signaling, and/or mediating complement- or cell-dependent lysis of tumor cells. More recent evidence suggests that TA-specific mAb can induce TA specific cytotoxic T-cell responses by enhancing TA uptake by dendritic cells and cross-priming of T cells. In this review, we briefly summarize the TA-specific mAbs that have received Food and Drug Administration approval. Next, we review the potential mechanisms underlying the therapeutic efficacy of TA-specific mAbs with emphasis on the induction of TA-specific cellular immune responses and their potential to contribute to the clinical efficacy of TA-specific mAb-based immunotherapy. Lastly, we discuss the potential negative effect of immune escape mechanisms on the clinical efficacy of TA-specific mAb-based immunotherapy. PMID- 20028763 TI - A group of genome-based biomarkers that add to a Kattan nomogram for predicting progression in men with high-risk prostate cancer. AB - PURPOSE: The three main treatment options for primary prostate cancer are surgery, radiation, and active surveillance. Surgical and radiation intervention for prostate cancer can be associated with significant morbidity. Therefore, accurate stratification predictive of outcome for prostate cancer patients is essential for appropriate treatment decisions. Nomograms that use clinical and pathologic variables are often used for risk prediction. Favorable outcomes exist even among men classified by nomograms as being at high risk of recurrence. EXPERIMENTAL DESIGN: Previously, we identified a set of DNA-based biomarkers termed Genomic Evaluators of Metastatic Prostate Cancer (GEMCaP) and have shown that they can predict risk of recurrence with 80% accuracy. Here, we examined the risk prediction ability of GEMCaP in a high-risk cohort and compared it to a Kattan nomogram. RESULTS: We determined that the GEMCaP genotype alone is comparable with the nomogram, and that for a subset of cases with negative lymph nodes improves upon it. CONCLUSION: Thus, GEMCaP shows promise for predicting unfavorable outcomes for negative lymph node high-risk cases, where the nomogram falls short, and suggests that addition of GEMCaP to nomograms may be warranted. PMID- 20028764 TI - Phase 1 study of aflibercept administered subcutaneously to patients with advanced solid tumors. AB - PURPOSE: To determine the maximum tolerated dose or maximal administered dose and pharmacokinetic and safety profiles of s.c. administered vascular endothelial growth factor Trap (aflibercept), a novel antiangiogenic agent. EXPERIMENTAL DESIGN: In this open-label, dose-escalation study, patients with advanced solid tumors were treated with subcutaneous doses of aflibercept at seven dose levels. Patients received a single dose of aflibercept and then underwent safety and pharmacokinetic assessments over the next 4 weeks. Patients then received weekly or biweekly treatment over the subsequent 6 weeks. Patients tolerating and benefiting could continue on aflibercept at the same dose and schedule until progression of disease. RESULTS: Thirty-eight patients received at least one dose of aflibercept. Maximum tolerated dose was not reached. Due to solubility/dosing limits with the subcutaneous formulation, 1,600 microg/kg/week was the maximal administered dose. The most common toxicities were proteinuria (37%), fatigue (32%), injection site reactions (18%), nausea (17%), myalgia and anorexia (16% each), hypertension (13%), and voice hoarseness (11%). Drug-related grade 3 to 4 toxicity was uncommon (7%) and reversible: dehydration, cerebral ischemia, proteinuria, hypertension, leukopenia, and pulmonary embolism. We identified dose proportional increases in plasma concentrations of aflibercept bound to vascular endothelial growth factor with a t(1/2) of 18 days. No antiaflibercept antibodies were detected. Stable disease was maintained for at least 10 weeks in 18 patients (47%), and 2 patients maintained on study for >1 year. CONCLUSION: Subcutaneous aflibercept was well tolerated and had manageable side effects. Its favorable pharmacokinetic profile and potential antitumor activity warrants further evaluation. PMID- 20028766 TI - Evaluating a marker's contribution to a nomogram: the GEMCaP example. AB - A postoperative nomogram developed in 1999 by Kattan et al. has been externally validated, but needs improvement. This tool predicts well but not perfectly. The Genomic Evaluators of Metastatic Prostate Cancer (GEMCaP) biomarkers hold promise for improving this tool; however, a larger data set that permits more involved analyses is needed. PMID- 20028765 TI - Vorinostat enhances the activity of temsirolimus in renal cell carcinoma through suppression of survivin levels. AB - PURPOSE: The mammalian target of rapamycin (mTOR) inhibitor temsirolimus has exhibited promising anticancer activity for the treatment of renal cell cancers (RCC). Survivin expression has been implicated in drug resistance and reducing its levels with the histone deacetylase (HDAC) inhibitor vorinostat may enhance the anticancer activity of temsirolimus. EXPERIMENTAL DESIGN: The sensitivity of RCC cell lines to the combination of temsirolimus and vorinostat was determined by measuring cell viability, clonogenic survival, and apoptosis. The effects of this combination on survivin levels were determined in vitro and in vivo. Survivin expression was silenced using small interfering RNA to evaluate its role in determining sensitivity to temsirolimus and vorinostat. The effect of the combination on angiogenesis was also determined in RCC xenograft models. RESULTS: Vorinostat synergistically improved the anticancer activity of temsirolimus in a panel of RCC cell lines in vitro and in two xenograft models in vivo. While each single agent led to a modest decrease in survivin levels, the combination dramatically reduced its expression, which correlated with an induction of apoptosis. Silencing survivin levels induced apoptosis and significantly improved the efficacy of temsirolimus and vorinostat. In addition, the temsirolimus/vorinostat combination led to a strong reduction in angiogenesis. CONCLUSIONS: Vorinostat augmented the anticancer activity of temsirolimus in both in vitro and in vivo models of RCC. The effectiveness of the combination was due to a decrease in survivin levels and corresponding induction of apoptosis, and enhanced inhibition of angiogenesis. Targeting survivin may be a promising therapeutic strategy to improve RCC therapy. PMID- 20028767 TI - Microarray-based response prediction in esophageal adenocarcinoma. AB - PURPOSE: In locally advanced (uT(3), N(+)) adenocarcinomas of the esophagus, neoadjuvant chemotherapy improves patient outcome. However, only a subgroup of patients responds. Therefore, in the present study, we evaluated whether the response to neoadjuvant chemotherapy can be predicted by a pretreatment tumor biopsy analysis. EXPERIMENTAL DESIGN: Biopsies of 47 patients with locally advanced (uT(3), N(+)) adenocarcinoma of the esophagus were obtained during primary staging. All patients underwent neoadjuvant chemotherapy with cisplatin, 5-fluorouracil, and leucovorin and subsequent resection of the esophagus. Biopsies were used for microarray analysis. The predominance of tumor cells within the specimens was >70%. Affymetrix U133 plus 2.0 gene chips with 54675 probe sets were used. A statistical comparison of patients responding to chemotherapy versus nonresponding patients was done. All patients were examined with immunohistology against Ephrin B3 receptor and Ki-67. RESULTS: A total of 86 genes were at least 2-fold differentially regulated comparing responding with nonresponding adenocarcinomas of the esophagus. The predominant genes encoded for the regulation of the cell cycle, transduction, translation, cell-cell interaction, cytoskeleton, and the signal transduction. The strongest difference was seen for the Ephrin B3 receptor. This result could be confirmed by immunhistology. A statistical significant correlation between the Ephrin B3 receptor, chemotherapy response, pathologic staging, and grading could be shown. CONCLUSIONS: There were significant differences in the gene profile between patients with adenocarcinoma of the esophagus responding to neoadjuvant chemotherapy compared with nonresponding patients. This suggests that it could be possible to characterize patients responding to chemotherapy even before starting the treatment using customized microarray analysis. PMID- 20028768 TI - Three DNA methylation epigenotypes in human colorectal cancer. AB - PURPOSE: Whereas the CpG island methylator phenotype (CIMP) in colorectal cancer associates with microsatellite instability (MSI)-high and BRAF-mutation(+), the existence of an intermediate-methylation subgroup associated with KRAS mutation(+) is controversial, and suitable markers for the subgroup have yet to be developed. Our aim is to clarify DNA methylation epigenotypes of colorectal cancer more comprehensively. EXPERIMENTAL DESIGN: To select new methylation markers on a genome-wide scale, we did methylated DNA immunoprecipitation-on-chip analysis of colorectal cancer cell lines and re-expression array analysis by 5 aza-2'-deoxycytidine/Trichostatin A treatment. Methylation levels were analyzed quantitatively in 149 colorectal cancer samples using matrix-assisted laser desorption/ionization-time-of-flight mass spectrometry. Colorectal cancer was epigenotyped by unsupervised two-way hierarchical clustering method. RESULTS: Among 1,311 candidate silencing genes, 44 new markers were selected and underwent quantitative methylation analysis in colorectal cancer samples together with 16 previously reported markers. Colorectal cancer was clustered into high-, intermediate-, and low-methylation epigenotypes. Methylation markers were clustered into two major groups: group 1 showing methylation in high-methylation epigenotype, and group 2 showing methylation in high- and intermediate methylation epigenotypes. A two-step marker panel deciding epigenotypes was developed with 95% accuracy: the 1st panel consisting of three group-1 markers (CACNA1G, LOX, SLC30A10) to extract high-methylation epigenotype, and the 2nd panel consisting of four group-2 markers (ELMO1, FBN2, THBD, HAND1) and SLC30A10 again to divide the remains into intermediate- and low-methylation epigenotypes. The high-methylation epigenotype correlated significantly with MSI-high and BRAF mutation(+) in concordance with reported CIMP. Intermediate-epigenotype significantly correlated with KRAS-mutation(+). KRAS-mutation(+) colorectal cancer with intermediate-methylation epigenotype showed significantly worse prognosis. CONCLUSIONS: Three methylation epigenotypes exist in colorectal cancer, and suitable classification markers have been developed. Intermediate methylation epigenotype with KRAS-mutation(+) correlated with worse prognosis. PMID- 20028769 TI - Expression of interleukin-1 receptor-associated kinase-1 in non-small cell lung carcinoma and preneoplastic lesions. AB - PURPOSE: To identify the pattern of interleukin-1 receptor-associated kinase (IRAK-1) protein expression in non-small cell lung carcinoma (NSCLC) and corresponding preneoplastic lesions. EXPERIMENTAL DESIGN: Archived tissue from NSCLC (adenocarcinoma and squamous cell carcinoma; n = 306) and adjacent bronchial epithelial specimens (n = 315) were analyzed for the immunohistochemical expression of IRAK-1, and the findings were correlated with patients' clinicopathologic features. Furthermore, we investigated the correlation between IRAK-1 expression and expression of NF-kappaB and IL-1alpha in tumor specimens. RESULTS: NSCLC tumors showed significantly higher cytoplasmic and lower nuclear IRAK-1 expression than normal epithelium. Squamous dysplasias had significantly higher cytoplasmic IRAK-1 expression than normal epithelium. In tumors, a significant positive correlation was detected between IRAK-1 expression (nuclear and cytoplasmic; P = 0.011) and IL-1alpha cytoplasmic expression (P < 0.0001). The correlation between the expression of the markers and patients' clinicopathologic features varied according to tumor histologic type and sex. High IRAK-1 cytoplasmic expression correlated with worse recurrence-free survival in women with NSCLC [hazard ratio (HR), 2.204; P = 0.033], but not in men. In adenocarcinoma, combined low level of expression of nuclear IRAK-1 and NF-kappaB correlated significantly with worse overall (HR, 2.485; P = 0.007) and recurrence free (HR, 3.058; P = 0.006) survivals in stage I/II patients. CONCLUSIONS: IRAK-1 is frequently expressed in NSCLC tissue specimens, and this expression is an early phenomenon in the sequential development of lung cancer. IRAK-1 is a novel inflammation-related marker and a potential target for lung cancer chemopreventive strategies. PMID- 20028770 TI - Targeting the cyclin E-Cdk-2 complex represses lung cancer growth by triggering anaphase catastrophe. AB - PURPOSE: Cyclin-dependent kinases (Cdk) and their associated cyclins are targets for lung cancer therapy and chemoprevention given their frequent deregulation in lung carcinogenesis. This study uncovered previously unrecognized consequences of targeting the cyclin E-Cdk-2 complex in lung cancer. EXPERIMENTAL DESIGN: Cyclin E, Cdk-1, and Cdk-2 were individually targeted for repression with siRNAs in lung cancer cell lines. Cdk-2 was also pharmacologically inhibited with the reversible kinase inhibitor seliciclib. Potential reversibility of seliciclib effects was assessed in washout experiments. Findings were extended to a large panel of cancer cell lines using a robotic-based platform. Consequences of cyclin E-Cdk-2 inhibition on chromosome stability and on in vivo tumorigenicity were explored as were effects of combining seliciclib with different taxanes in lung cancer cell lines. RESULTS: Targeting the cyclin E-Cdk-2 complex, but not Cdk-1, resulted in marked growth inhibition through the induction of multipolar anaphases triggering apoptosis. Treatment with the Cdk-2 kinase inhibitor seliciclib reduced lung cancer formation in a murine syngeneic lung cancer model and decreased immunohistochemical detection of the proliferation markers Ki-67 and cyclin D1 in lung dysplasia spontaneously arising in a transgenic cyclin E-driven mouse model. Combining seliciclib with a taxane resulted in augmented growth inhibition and apoptosis in lung cancer cells. Pharmacogenomic analysis revealed that lung cancer cell lines with mutant ras were especially sensitive to seliciclib. CONCLUSIONS: Induction of multipolar anaphases leading to anaphase catastrophe is a previously unrecognized mechanism engaged by targeting the cyclin E-Cdk-2 complex. This exerts substantial antineoplastic effects in the lung. PMID- 20028771 TI - Phase I study of the angiogenesis inhibitor BIBF 1120 in patients with advanced solid tumors. AB - PURPOSE: BIBF 1120 is an oral, potent angiokinase inhibitor targeting receptors of the vascular endothelial growth factors, platelet-derived growth factors, and fibroblast growth factors. This phase I, accelerated titration study assessed the maximum tolerated dose, safety, pharmacokinetics, and pharmacodynamic effects of BIBF 1120. PATIENTS AND METHODS: Sixty-one patients with advanced cancers received BIBF 1120 in successive cohorts. Twenty-five received 50 to 450 mg once daily and 36 received 150 to 300 mg twice daily in 4-week treatment courses interspersed by 1 week of washout. Dynamic contrast-enhanced magnetic resonance imaging assessed antiangiogenic effect in 42 patients. RESULTS: Most frequent BIBF 1120-related adverse events were mostly mild to moderate (Common Toxicity Criteria grade 1-2) nausea (68.9%), vomiting (45.9%), and diarrhea (44.3%). The majority of dose-limiting adverse events of Common Toxicity Criteria grade 3 or 4 were reversible liver enzyme elevations. The maximum tolerated dose was 250 mg of BIBF 1120 for once and twice daily dosing. BIBF 1120 was absorbed moderately fast (t(max) = 1-3 hours at steady state), with no deviation from dose linearity and no decrease of exposure over time. The gMean terminal half-life was from 13 to 19 hours. One complete and two partial responses occurred in patients with renal cell cancer (n = 2) and colorectal cancer (n = 1). Dynamic contrast-enhanced magnetic resonance imaging showed a significant reduction in tumor blood flow in 55% of evaluable patients. CONCLUSIONS: BIBF 1120 dosed continuously displayed a favorable safety and pharmacokinetics profile, and first efficacy signals were observed. Twice daily dosing permitted increased drug exposure without additional toxicity. Two hundred milligrams BIBF 1120 twice daily is the recommended dose for phase II monotherapy studies. PMID- 20028772 TI - CCR7 regulates lymphocyte egress and recirculation through body cavities. AB - T and B lymphocytes recirculate among blood, lymph, and extralymphoid tissues to ensure immune surveillance and the establishment of self-tolerance. The underlying mechanisms regulating homeostatic lymphocyte recirculation through body cavities are not fully understood. Here, we demonstrate that the homeostatic chemokine receptor CCR7 regulates homeostatic recirculation of lymphocytes through body cavities. CCR7 deficiency results in massive accumulation of CD4(+) and CD8(+) T cells and B-2 B cells in the peritoneal and pleural cavities. The increase in B-2 B and T lymphocytes is not associated with an altered maturation and/or activation status of these cells. Mechanistically, an increase in peritoneal lymphocyte numbers is caused by impaired egress of CCR7-deficient lymphocytes from body cavities. These results establish that CCR7 plays a crucial role in lymphocyte exit from the PerC. PMID- 20028773 TI - Liver X receptor agonist treatment reduced splanchnic ischemia and reperfusion injury. AB - LXR is another member of the superfamily of nuclear hormone receptors that heterodimerizes with RXR and regulates the intracellular levels of cholesterol through gene induction of enzymes and proteins involved in the cholesterol metabolism and transport. LXR ligands inhibit the gene expression of proinflammatory mediators in immunostimulated macrophages; in vivo studies have shown that activation of LXR reduces the inflammatory response in a murine model of contact dermatitis and atherosclerosis. No reports have addressed a role for LXRs in pathophysiology of intestinal ischemia. The aim of this study was to investigate the effects of T0901317, a potent LXR ligand, in a mouse model of SAO shock, which was induced by clamping the superior mesenteric artery and the celiac trunk, resulting in a total occlusion of these arteries for 30 min. After this period of occlusion, the clamps were removed. Mice were killed at 60 min after reperfusion. This study provides the evidence that T0901317, LXR agonist, modulates: the development of SAO shock; the infiltration of the tissue with PMNs; the expression of TNF-alpha and IL-1beta; the nitration of tyrosine residues; NF-kappaB expression; the MAPK phosphorylation (ERK, JNK, and p38); FasL; apoptosis; Bax and Bcl-2 expression; and the degree of tissue injury caused by SAO shock. Our results imply that LXR agonists may be useful in the therapy of inflammation. PMID- 20028774 TI - Macrophages from BALB/c and CBA/Ca mice differ in their cellular responses to Streptococcus pneumoniae. AB - In a mouse model of pneumonia caused by Streptococcus pneumoniae, differences in the timing and vigor of the host inflammatory response have been associated with susceptibility to invasive disease. BALB/c and CBA/Ca mice are known to be resistant and susceptible to acute pneumococcal disease, respectively. In this study, we have demonstrated that BMM from BALB/c and CBA/Ca mice differ in their expression and regulation of TLR9 in response to S. pneumoniae. We have also shown that BMM from CBA/Ca mice failed to fully activate p38, NF-kappaB, and ERK 1/2 signaling pathways, resulting in reduced secretion of TNF-alpha and CCL5 in response to this pathogen. In addition, we have established that S. pneumoniae induced significant cell death in BMM from CBA/Ca mice. These findings indicate that variations between the two strains are likely to reflect differences in macrophage responses to the pathogen. PMID- 20028775 TI - The T cell receptor-mediated phosphorylation of Pyk2 tyrosines 402 and 580 occurs via a distinct mechanism than other receptor systems. AB - The tyrosine kinase Pyk2 is vital for integrating receptor-mediated signals controlling adhesion and motility in neuronal, epithelial, and hematopoietic cell types. In T cells, the stimulation of the TCR and costimulatory, chemokine, cytokine, and integrin receptors leads to the phosphorylation of Pyk2 and the induction of its catalytic activity. However, our understanding of the mechanism of the TCR-induced, site-specific phosphorylation of this kinase is incomplete and contradictory. To address this issue, the role of individual signaling pathways in the phosphorylation of Pyk2 tyrosines 402 and 580 upon TCR activation was assessed in human T cells. In contrast to other receptor systems, the TCR induced phosphorylation of Pyk2 tyrosines 402 and 580 was dependent on the Src family kinases, Fyn or Lck. Interestingly, the TCR-mediated phosphorylation of Pyk2 tyrosines 402 and 580 did not require Ca(2+) influx, ZAP-70 activation, actin cytoskeleton rearrangement, or PI3K function. These observations are different than other receptor systems, which require the induction of one or more of these pathways. Together, these data have defined more fully the mechanism for the TCR-induced phosphorylation of specific sites on Pyk2, suggesting that the TCR has a distinct pathway for the activation of Pyk2 compared with other receptor systems. PMID- 20028776 TI - Health and disease in people over 85. PMID- 20028777 TI - Health and disease in 85 year olds: baseline findings from the Newcastle 85+ cohort study. AB - OBJECTIVES: The Newcastle 85+ Study aims to systematically study the clinical, biological, and psychosocial attributes of an unselected cohort of 85 year olds and to examine subsequent health trajectories as the cohort ages; health at baseline is reported. DESIGN: Cross sectional analysis of baseline data from a cohort study. SETTING: Newcastle upon Tyne and North Tyneside primary care trusts, United Kingdom. PARTICIPANTS: 1042 people born in 1921 and registered with the participating general practices. MAIN OUTCOME MEASURES: Detailed health assessment and review of general practice records (disease, medication, and use of general practice services); participants could decline elements of the protocol. RESULTS: Of the 1453 eligible people, 851 (58.6%) were recruited to health assessment plus record review, 188 (12.9%) to record review only, and 3 (0.2%) to health assessment only. Data from record review are reported on a maximum of 1030 and from health assessment on a maximum of 853; individual denominators differ owing to withdrawal and missing values. Of the health assessment sample (n=853), 62.1% (n=530) were women and 10.4% (n=89) were in institutional care. The most prevalent diseases were hypertension (57.5%, 592/1030) and osteoarthritis (51.8%, 534/1030). Moderate or severe cognitive impairment was present in 11.7% (96/824) of participants, severe or profound urinary incontinence in 21.3% (173/813), hearing impairment in 59.6% (505/848), and visual impairment in 37.2% (309/831). Health assessment identified participants with possible disease but without a previous diagnosis in their medical record for hypertension (25.1%, 206/821), ischaemic heart disease (12.6%, 99/788), depression (6.9%, 53/772), dementia (6.7%, 56/840), and atrial fibrillation (3.8%, 30/788). Undiagnosed diabetes mellitus and thyroid disease were rare (1%, 7/717 and 6/762, respectively). A median of 3 (interquartile range 1-8) activities of daily living were undertaken with difficulty. Overall, 77.6% (646/832) of participants rated their health compared with others of the same age as good, very good, or excellent. High contact rates in the previous year with general practitioners (93.8%, 960/1024) were recorded. Women had significantly higher disease counts (medians: women 5, men 4; P=0.033) and disability scores (medians: women 4, men 2; P=0.0006) than men, but were less likely to have attended outpatient clinics in the previous three months (women 29% (150/524), men 37% (118/320), odds ratio 0.7, 95% confidence interval 0.5 to 0.9). CONCLUSIONS: This large cohort of 85 year olds showed good levels of both self rated health and functional ability despite significant levels of disease and impairment. Hypertension, ischaemic heart disease, atrial fibrillation, depression, and dementia may be underdiagnosed. Notable differences were found between the sexes: women outnumbered men and had more disease and disability. PMID- 20028778 TI - The association between BMI and mortality using offspring BMI as an indicator of own BMI: large intergenerational mortality study. AB - OBJECTIVES: To obtain valid estimates of the association between body mass index (BMI) and mortality by using offspring BMI as an instrumental variable for own BMI. DESIGN: Cohort study based on record linkage, with 50 years of follow-up for mortality. Associations of offspring BMI with all cause and cause specific maternal and paternal mortality were estimated as hazard ratios per standard deviation of offspring BMI. SETTING: A large intergenerational prospective population based database covering the general population of Sweden. PARTICIPANTS: More than one million Swedish parent-son pairs. RESULTS: The final dataset analysed contained information on 1 018 012 mother-son pairs (122 677 maternal deaths) and 1 004 617 father-son pairs (242 126 paternal deaths). For some causes of death, the patterns of associations between offspring BMI and mortality were similar to those seen for own BMI and mortality in previous studies. Parental mortality from diabetes, coronary heart disease, and kidney cancer had the strongest positive associations with offspring BMI (for example, hazard ratio (HR) for coronary heart disease per standard deviation increase in offspring BMI for mothers 1.15, 95% CI 1.14 to 1.17 and for fathers 1.10, 1.09 to 1.11). However, in contrast to the inverse association of own BMI with lung cancer and respiratory disease mortality seen in other studies, there was a positive association between offspring BMI and lung cancer mortality in mothers (1.12, 1.09 to 1.15) and fathers (1.03, 1.02 to 1.05) and between offspring BMI and respiratory mortality in mothers (1.05, 1.02 to 1.08) and fathers (1.02, 1.00 to 1.04). Associations of own BMI and offspring BMI with all cause, cardiovascular disease related, and non-cardiovascular disease related mortality were compared in a subset of father-son pairs (n=72 815). When offspring BMI was used as an instrumental variable for paternal BMI, the causal association between BMI and paternal cardiovascular disease mortality (HR per standard deviation of BMI 1.82, 95% CI 1.17 to 2.83) was stronger than that indicated by the directly observed association between own BMI and cardiovascular disease mortality (1.45, 1.31 to 1.61). CONCLUSIONS: Use of offspring BMI as a predictor of own BMI, a technique that avoids problems of reverse causality, suggests that positive associations of BMI with all cause and cardiovascular mortality may be underestimated in conventional observational studies. Use of offspring BMI instead of own BMI in analyses of respiratory disease and lung cancer mortality, for which previous studies have reported consistent and strong inverse associations with own BMI, suggests that such studies have overstated the apparent adverse consequences of lower BMI with respect to these outcomes. PMID- 20028779 TI - Screening for postnatal depression in primary care: cost effectiveness analysis. AB - OBJECTIVE: To evaluate the cost effectiveness of routine screening for postnatal depression in primary care. DESIGN: Cost effectiveness analysis with a decision model of alternative methods of screening for depression, including standardised postnatal depression and generic depression instruments. The performance of screening instruments was derived from a systematic review and bivariate meta analysis at a range of instrument cut points; estimates of other relevant parameters were derived from literature sources and relevant databases. A decision tree considered the full treatment pathway from the possible onset of postnatal depression through identification, treatment, and possible relapse. SETTING: Primary care. PARTICIPANTS: A hypothetical population of women assessed for postnatal depression either via routine care only or supplemented by use of formal identification methods six weeks postnatally, as recommended in recent guidelines. MAIN OUTCOME MEASURES: Costs expressed in 2006-7 prices and impact on health outcomes expressed in terms of quality adjusted life years (QALYs). The time horizon of the analysis was one year. RESULTS: The routine application of either postnatal or general depression questionnaires did not seem to be cost effective compared with routine care only. The Edinburgh postnatal depression scale (at a cut point of 16) had an incremental cost effectiveness ratio (ICER) of pound 41,103 (euro 45,398, $67,130) per QALY compared with routine care only. The ICER for all other strategies ranged from pound 49,928 to pound 272,463 per QALY versus routine care only, while the probability that no formal identification strategy was cost effective was 88% (59%) at a cost effectiveness threshold of pound 20,000 ( pound 30,000) per QALY. While sensitivity analysis indicated that the cost of managing incorrectly identified depression (false positive result) was an important driver of the model, formal identification approaches did not seem to be cost effective at any feasible estimate of this cost. CONCLUSIONS: Formal identification methods for postnatal depression do not seem to represent value for money for the NHS. The major determinant of cost effectiveness seems to be the potential additional costs of managing women incorrectly diagnosed as depressed. Formal identification methods for postnatal depression do not currently satisfy the National Screening Committee's criteria for the adoption of a screening strategy as part of national health policy. PMID- 20028780 TI - Genetic analysis of extensively drug-resistant Mycobacterium tuberculosis strains in Lisbon, Portugal. AB - OBJECTIVES: Extensively drug-resistant (XDR) tuberculosis (TB) threatens the global control of TB worldwide. Lisbon has a high XDR-TB rate [50% of the multidrug-resistant tuberculosis (MDR-TB)], which is mainly associated with Lisboa family strains. Few studies have addressed the identification of mutations associated with resistance to second-line injectable drugs, and the relative frequency of such mutations varies geographically. The aim of this study was to characterize the genetic changes associated with the high number of XDR-TB cases in Lisbon. METHODS: In the present study we analysed 26 XDR-TB clinical isolates. The gyrA, tlyA and rrs genes were screened for mutations that could be responsible for resistance to fluoroquinolones and second-line injectable drugs. Moreover, the strains under analysis were also genotyped by MIRU-VNTR ('mycobacterial interspersed repetitive unit-variable number of tandem repeats'). RESULTS: The mutational analysis identified the most frequent mutations in the resistance-associated genes: S91P in gyrA (42.3%); A1401G in rrs (30.8%); and Ins755GT in tlyA (42.3%). The occurrence of mutations in rrs was associated with the non-occurrence of mutations in tlyA. The genotypic analysis revealed that the strains were highly clonal, belonging to one of two MIRU-VNTR clusters, with the largest belonging to the Lisboa family. Association between mutations in gyrA and rrs or tlyA was verified. CONCLUSIONS: The association of specific mutations highlighted the strains' high clonality and indicates recent XDR-TB transmission. In addition, the identification of the most frequent resistance-associated mutations will be invaluable in applying XDR-TB molecular detection tests in the region in the near future. PMID- 20028781 TI - Will Swedish healthcare reforms affect equity? PMID- 20028782 TI - Emergency sedation in children. Utility of low dose ketamine. PMID- 20028786 TI - Commerce and malnutrition. Breast feeding in Bangladesh: time for government action. PMID- 20028790 TI - The genetic and metabolic signature of oncocytic transformation implicates HIF1alpha destabilization. AB - We previously showed that disruptive complex I mutations in mitochondrial DNA are the main genetic hallmark of oncocytic tumors of the thyroid and kidney. We here report a high frequency of homoplasmic disruptive mutations in a large panel of oncocytic pituitary and head-and-neck tumors. The presence of such mutations implicates disassembly of respiratory complex I in vivo which in turn contributes to the inability of oncocytic tumors to stabilize HIF1alpha and to display pseudo hypoxia. By utilizing transmitochondrial cytoplasmic hybrids (cybrids), we induced the shift to homoplasmy of a truncating mutation in the mitochondria coded MTND1 gene. Such shift is associated with a profound metabolic impairment leading to the imbalance of alpha-ketoglutarate and succinate, the Krebs cycle metabolites which are the main responsible for HIF1alpha stabilization. We conclude that the main hallmarks of oncocytic transformation, namely the occurrence of homoplasmic disruptive mutations and complex I disassembly, may explain the benign nature of oncocytic neoplasms through lack of HIF1alpha stabilization. PMID- 20028791 TI - Disease mutations in Rab7 result in unregulated nucleotide exchange and inappropriate activation. AB - Rab GTPases are molecular switches that orchestrate vesicular trafficking, maturation and fusion by cycling between an active, GTP-bound form, and an inactive, GDP-bound form. The activity cycle is coupled to GTP hydrolysis and is tightly controlled by regulatory proteins. Missense mutations of the GTPase Rab7 cause a dominantly inherited axonal degeneration known as Charcot-Marie-Tooth type 2B through an unknown mechanism. We present the 2.8 A crystal structure of GTP-bound L129F mutant Rab7 which reveals normal conformations of the effector binding regions and catalytic site, but an alteration to the nucleotide binding pocket that is predicted to alter GTP binding. Through extensive biochemical analysis, we demonstrate that disease-associated mutations in Rab7 do not lead to an intrinsic GTPase defect, but permit unregulated nucleotide exchange leading to both excessive activation and hydrolysis-independent inactivation. Consistent with augmented activity, mutant Rab7 shows significantly enhanced interaction with a subset of effector proteins. In addition, dynamic imaging demonstrates that mutant Rab7 is abnormally retained on target membranes. However, we show that the increased activation of mutant Rab7 is counterbalanced by unregulated, GTP hydrolysis-independent membrane cycling. Notably, disease mutations are able to rescue the membrane cycling of a GTPase-deficient mutant. Thus, we demonstrate that disease mutations uncouple Rab7 from the spatial and temporal control normally imposed by regulatory proteins and cause disease not by a gain of novel toxic function, but by misregulation of native Rab7 activity. PMID- 20028801 TI - Ontogeny of the oestrogen receptors ESR1 and ESR2 during gonadal development in the tammar wallaby, Macropus eugenii. AB - Oestrogen has wide ranging effects in development mediated mainly via the two oestrogen receptors, alpha (ESR1, also known as ERalpha) and beta (ESR2, also known as ERbeta). Oestrogen is the key factor that directs the indifferent gonad to become an ovary in many non-mammalian vertebrates. Oestrogen is not required for early ovarian differentiation in mammals but can disrupt normal testicular development in eutherians. Surprisingly, exogenous oestrogen can cause sex reversal of an XY gonad in two marsupials, the North American opossum and the tammar wallaby. To understand the mechanism by which oestrogen induces sex reversal, we characterised the genes for ESR1 and ESR2 and examined their expression during gonadal differentiation in the tammar wallaby, Macropus eugenii. Both receptors were expressed in the somatic cells and germ cells of the indifferent gonad in both XX and XY foetuses throughout all stages of development, and persisted in these cells into adulthood. ERs were also present in many other tissues including kidney, pituitary and mammary gland. ER mRNA was not significantly altered by exogenous oestrogen in cultured XY gonads but the receptors translocated to the nucleus in its presence. These findings confirm that there is conserved expression of the ERs in the indifferent gonad despite the lack of available ligand during early gonadal development. The receptors can respond to exogenous estrogen at this early stage and are capable of transducing signals in the early mammalian gonad. However, the selective forces that maintained conserved ER expression in this tissue remain unknown. PMID- 20028792 TI - Mistargeting of SH3TC2 away from the recycling endosome causes Charcot-Marie Tooth disease type 4C. AB - Mutations in the functionally uncharacterized protein SH3TC2 are associated with the severe hereditary peripheral neuropathy, Charcot-Marie-Tooth disease type 4C (CMT4C). Similarly, to other proteins mutated in CMT, a role for SH3TC2 in endocytic membrane traffic has been previously proposed. However, recent descriptions of the intracellular localization of SH3TC2 are conflicting. Furthermore, no clear functional pathogenic mechanisms have so far been proposed to explain why both nonsense and missense mutations in SH3TC2 lead to similar clinical phenotypes. Here, we describe our intracellular localization studies, supported by biochemical and functional data, using wild-type and mutant SH3TC2. We show that wild-type SH3TC2 targets to the intracellular recycling endosome by associating with the small GTPase, Rab11, which is known to regulate the recycling of internalized membrane and receptors back to the plasma membrane. Furthermore, we demonstrate that SH3TC2 interacts preferentially with the GTP bound form of Rab11, identifying SH3TC2 as a novel Rab11 effector. Of clinical pathological relevance, all SH3TC2 constructs harbouring disease-causing mutations are shown to be unable to associate with Rab11 with consequent loss of recycling endosome localization. Moreover, we show that wild-type SH3TC2, but not mutant SH3TC2, influences transferrin receptor dynamics, consistent with a functional role on the endocytic recycling pathway. Our data therefore implicate mistargeting of SH3TC2 away from the recycling endosome as the fundamental molecular defect that leads to CMT4C. PMID- 20028802 TI - Illness deception and work: incidence, manifestations and detection. AB - BACKGROUND: Illness deception (ID) may be difficult to detect. Objective techniques that improve a doctor's skills of detection will have benefits for the health economy. AIMS: To ascertain the incidence of ID in a general occupational medicine clinic, to give examples by way of case vignettes how it may manifest and to compare the variability of maximal grip strength measurements of patients with suspected simulated weakness of grip with controls. METHODS: Four hundred consecutive new patient referrals were examined by the author and those who met Diagnostic and Statistical Manual-IV criteria for factitious disorder (FD) or malingering identified. Those with suspected simulated weakness of grip were asked to perform three consecutive maximal grips with a Jamar hand dynamometer. One hundred normal subjects and 100 patients with rheumatoid arthritis (RA) of the hands were similarly tested. RESULTS: Thirty-two of 400 (8%) patients exhibited behaviour in keeping with ID (29 malingering and 3 FD). Cases included simulated hand-arm vibration syndrome, occupational asthma, deafness and weakness of a limb. The median and 90th percentile for coefficient of variation (CV) of three consecutive maximal grip strengths for normal subjects and patients with RA were 5.2, 10.5%; 5.4, 14.5% right and 4.5, 10.2%; 6.0, 14.4% left hand, respectively. The CV of the six patients who simulated weakness of grip was from 17.3 to 37.8%. CONCLUSIONS: ID is relatively common in occupational medical practice and multiform in its manifestations. Simulated weakness of grip should be suspected when consecutive declared maximal grip strengths are inexplicably submaximal for age and sex and highly variable in force. PMID- 20028803 TI - The Pseudomonas aeruginosa magnesium transporter MgtE inhibits transcription of the type III secretion system. AB - Pseudomonas aeruginosa is an opportunistic pathogen that causes life-long pneumonia in individuals with cystic fibrosis (CF). These long-term infections are maintained by bacterial biofilm formation in the CF lung. We have recently developed a model of P. aeruginosa biofilm formation on cultured CF airway epithelial cells. Using this model, we discovered that mutation of a putative magnesium transporter gene, called mgtE, led to increased cytotoxicity of P. aeruginosa toward epithelial cells. This altered toxicity appeared to be dependent upon expression of the type III secretion system (T3SS). In this study, we found that mutation of mgtE results in increased T3SS gene transcription. Through epistasis analyses, we discovered that MgtE influences the ExsE-ExsC-ExsD ExsA gene regulatory system of T3SS by either directly or indirectly inhibiting ExsA activity. While variations in calcium levels modulate T3SS gene expression in P. aeruginosa, we found that addition of exogenous magnesium did not inhibit T3SS activity. Furthermore, mgtE variants that were defective for magnesium transport could still complement the cytotoxicity effect. Thus, the magnesium transport function of MgtE does not fully explain the regulatory effects of MgtE on cytotoxicity. Overall, our results indicate that MgtE modulates expression of T3SS genes. PMID- 20028804 TI - A tolC mutant of Francisella tularensis is hypercytotoxic compared to the wild type and elicits increased proinflammatory responses from host cells. AB - The highly infectious bacterium Francisella tularensis is a facultative intracellular pathogen and the causative agent of tularemia. TolC, which is an outer membrane protein involved in drug efflux and type I protein secretion, is required for the virulence of the F. tularensis live vaccine strain (LVS) in mice. Here, we show that an LVS DeltatolC mutant colonizes livers, spleens, and lungs of mice infected intradermally or intranasally, but it is present at lower numbers in these organs than in those infected with the parental LVS. For both routes of infection, colonization by the DeltatolC mutant is most severely affected in the lungs, suggesting that TolC function is particularly important in this organ. The DeltatolC mutant is hypercytotoxic to murine and human macrophages compared to the wild-type LVS, and it elicits the increased secretion of proinflammatory chemokines from human macrophages and endothelial cells. Taken together, these data suggest that TolC function is required for F. tularensis to inhibit host cell death and dampen host immune responses. We propose that, in the absence of TolC, F. tularensis induces excessive host cell death, causing the bacterium to lose its intracellular replicative niche. This results in lower bacterial numbers, which then are cleared by the increased innate immune response of the host. PMID- 20028805 TI - AatA is a novel autotransporter and virulence factor of avian pathogenic Escherichia coli. AB - Autotransporters (AT) are widespread in Gram-negative bacteria, and many of them are involved in virulence. An open reading frame (APECO1_O1CoBM96) encoding a novel AT was located in the pathogenicity island of avian pathogenic Escherichia coli (APEC) O1's virulence plasmid, pAPEC-O1-ColBM. This 3.5-kb APEC autotransporter gene (aatA) is predicted to encode a 123.7-kDa protein with a 25 amino-acid signal peptide, an 857-amino-acid passenger domain, and a 284-amino acid beta domain. The three-dimensional structure of AatA was also predicted by the threading method using the I-TASSER online server and then was refined using four-body contact potentials. Molecular analysis of AatA revealed that it is translocated to the cell surface, where it elicits antibody production in infected chickens. Gene prevalence analysis indicated that aatA is strongly associated with E. coli from avian sources but not with E. coli isolated from human hosts. Also, AatA was shown to enhance adhesion of APEC to chicken embryo fibroblast cells and to contribute to APEC virulence. PMID- 20028806 TI - Influence of mannan and glucan on complement activation and C3 binding by Candida albicans. AB - The complement system is important for host resistance to hematogenously disseminated candidiasis. However, modulation of complement activation by cell wall components of Candida albicans has not been characterized. Although intact yeast display mannan on the surface, glucan, typically located in the interior, becomes exposed during C. albicans infection. We show here the distinct effects of mannan and glucan on complement activation and opsonophagocytosis. Previous studies showed that intact cells are resistant to initiation of complement activation through the alternative pathway, and antimannan antibody reverses this resistance via an Fc-independent mechanism. The present study shows that this mannan-dependent resistance can be overcome by periodate-borohydride conversion of mannose polysaccharides to polyalcohols; cells treated with periodate borohydride initiate the alternative pathway without the need for antibody. These observations identify an inhibitory role for intact mannan in complement activation. Next, removal of the surface-displayed mannan by acid treatment of periodate-borohydride cells exposes glucan. Glucan-displaying cells or purified beta-glucan initiate the alternative pathway when incubated with the purified proteins of the alternative pathway alone, suggesting that C. albicans glucan is a natural activator of the alternative pathway. Finally, ingestion of mannan displaying cells by human neutrophils requires anti-mannan antibody, whereas ingestion of glucan-displaying cells requires complement. These results demonstrate a contrasting requirement of natural antibody and complement for opsonophagocytosis of C. albicans cells displaying mannan or glucan. Thus, differential surface expression of mannan and glucan may influence recognition of C. albicans by the complement system. PMID- 20028807 TI - A foreign protein incorporated on the Tip of T3 pili in Lactococcus lactis elicits systemic and mucosal immunity. AB - The use of Lactococcus lactis to deliver a chosen antigen to the mucosal surface has been shown to elicit an immune response in mice and is a possible method of vaccination in humans. The recent discovery on Gram-positive bacteria of pili that are covalently attached to the bacterial surface and the elucidation of the residues linking the major and minor subunits of such pili suggests that the presentation of an antigen on the tip of pili external to the surface of L. lactis might constitute a successful vaccine strategy. As a proof of principle, we have fused a foreign protein (the Escherichia coli maltose-binding protein) to the C-terminal region of the native tip protein (Cpa) of the T3 pilus derived from Streptococcus pyogenes and expressed this fusion protein (MBP*) in L. lactis. We find that MBP* is incorporated into pili in this foreign host, as shown by Western blot analyses of cell wall proteins and by immunogold electron microscopy. Furthermore, since the MBP* on these pili retains its native biological activity, it appears to retain its native structure. Mucosal immunization of mice with this L. lactis strain expressing pilus-linked MBP* results in production of both a systemic and a mucosal response (IgG and IgA antibodies) against the MBP antigen. We suggest that this type of mucosal vaccine delivery system, which we term UPTOP (for unhindered presentation on tips of pili), may provide an inexpensive and stable alternative to current mechanisms of immunization for many serious human pathogens. PMID- 20028808 TI - Molecular characterization of the Dot/Icm-translocated AnkH and AnkJ eukaryotic like effectors of Legionella pneumophila. AB - Although most Dot/Icm-translocated effectors of Legionella pneumophila are not required for intracellular proliferation, the eukaryotic-like ankyrin effectors, AnkH and AnkJ are required for intracellular proliferation. In this report, we show that the IcmSW chaperones are essential for translocation of AnkJ but not AnkH. The 10 C-terminal residues and the ANK domains of AnkH and AnkJ are required for translocation. Our data indicate that the two ANK domains of AnkH are critical domains required for the function of the effector in intracellular replication of L. pneumophila. The ankH and ankJ mutants are severely defective in intrapulmonary proliferation in mice. Expression of AnkH and AnkJ fusions within HEK293 cells show a punctuate distribution in the cytosol but no association with endocytic vesicles, the Golgi apparatus or the endoplasmic reticulum. Interestingly, the defect in intracellular proliferation of the ankH or ankJ mutants is rescued in HEK293 cells expressing the respective protein. We conclude that AnkH and AnkJ are effectors translocated by the Dot/Icm system by distinct mechanisms and modulate distinct cytosolic processes in the host cell. PMID- 20028810 TI - Phosphoglucomutase of Yersinia pestis is required for autoaggregation and polymyxin B resistance. AB - Yersinia pestis, the causative agent of plague, autoaggregates within a few minutes of cessation of shaking when grown at 28 degrees C. To identify the autoaggregation factor of Y. pestis, we performed mariner-based transposon mutagenesis. Autoaggregation-defective mutants from three different pools were identified, each with a transposon insertion at a different position within the gene encoding phosphoglucomutase (pgmA; y1258). Targeted deletion of pgmA in Y. pestis KIM5 also resulted in loss of autoaggregation. Given the previously defined role for phosphoglucomutase in antimicrobial peptide resistance in other organisms, we tested the KIM5 DeltapgmA mutant for antimicrobial peptide sensitivity. The DeltapgmA mutant displayed >1,000-fold increased sensitivity to polymyxin B compared to the parental Y. pestis strain, KIM5. This sensitivity is not due to changes in lipopolysaccharide (LPS) since the LPSs from both Y. pestis KIM5 and the DeltapgmA mutant are identical based on a comparison of their structures by mass spectrometry (MS), tandem MS, and nuclear magnetic resonance analyses. Furthermore, the ability of polymyxin B to neutralize LPS toxicity was identical for LPS purified from both KIM5 and the DeltapgmA mutant. Our results indicate that increased polymyxin B sensitivity of the DeltapgmA mutant is due to changes in surface structures other than LPS. Experiments with mice via the intravenous and intranasal routes did not demonstrate any virulence defect for the DeltapgmA mutant, nor was flea colonization or blockage affected. Our findings suggest that the activity of PgmA results in modification and/or elaboration of a surface component of Y. pestis responsible for autoaggregation and polymyxin B resistance. PMID- 20028809 TI - QseC mediates Salmonella enterica serovar typhimurium virulence in vitro and in vivo. AB - The autoinducer-3 (AI-3)/epinephrine (Epi)/norepinephrine (NE) interkingdom signaling system mediates chemical communication between bacteria and their mammalian hosts. The three signals are sensed by the QseC histidine kinase (HK) sensor. Salmonella enterica serovar Typhimurium is a pathogen that uses HKs to sense its environment and regulate virulence. Salmonella serovar Typhimurium invades epithelial cells and survives within macrophages. Invasion of epithelial cells is mediated by the type III secretion system (T3SS) encoded in Salmonella pathogenicity island 1 (SPI-1), while macrophage survival and systemic disease are mediated by the T3SS encoded in SPI-2. Here we show that QseC plays an important role in Salmonella serovar Typhimurium pathogenicity. A qseC mutant was impaired in flagellar motility, in invasion of epithelial cells, and in survival within macrophages and was attenuated for systemic infection in 129x1/SvJ mice. QseC acts globally, regulating expression of genes within SPI-1 and SPI-2 in vitro and in vivo (during infection of mice). Additionally, dopamine beta hydroxylase knockout (Dbh(-)(/)(-)) mice that do not produce Epi or NE showed different susceptibility to Salmonella serovar Typhimurium infection than wild type mice. These data suggest that the AI-3/Epi/NE signaling system is a key factor during Salmonella serovar Typhimurium pathogenesis in vitro and in vivo. Elucidation of the role of this interkingdom signaling system in Salmonella serovar Typhimurium should contribute to a better understanding of the complex interplay between the pathogen and the host during infection. PMID- 20028811 TI - Fibrinogen regulates the cytotoxicity of mycobacterial trehalose dimycolate but is not required for cell recruitment, cytokine response, or control of mycobacterial infection. AB - During inflammatory responses and wound healing, the conversion of soluble fibrinogen to fibrin, an insoluble extracellular matrix, long has been assumed to create a scaffold for the migration of leukocytes and fibroblasts. Previous studies concluded that fibrinogen is a necessary cofactor for mycobacterial trehalose 6,6'-dimycolate-induced responses, because trehalose dimycolate-coated beads, to which fibrinogen was adsorbed, were more inflammatory than those to which other plasma proteins were adsorbed. Herein, we investigate roles for fibrin(ogen) in an in vivo model of mycobacterial granuloma formation and in infection with Mycobacterium tuberculosis, the causative agent of tuberculosis. In wild-type mice, the subcutaneous injection of trehalose dimycolate-coated polystyrene microspheres, suspended within Matrigel, elicited a pyogranulomatous response during the course of 12 days. In fibrinogen-deficient mice, neutrophils were recruited but a more suppurative lesion developed, with the marked degradation and disintegration of the matrix. Compared to that in wild-type mice, the early formation of granulation tissue in fibrinogen-deficient mice was edematous, hypocellular, and disorganized. These deficiencies were complemented by the addition of exogenous fibrinogen. The absence of fibrinogen had no effect on cell recruitment or cytokine production in response to trehalose dimycolate, nor was there a difference in lung histopathology or overall bacterial burden in mice infected with Mycobacterium tuberculosis. In this model, fibrin(ogen) was not required for cell recruitment, cytokine response, or response to infection, but it promoted granulation tissue formation and suppressed leukocyte necrosis. PMID- 20028812 TI - Extracts of the rat tapeworm, Hymenolepis diminuta, suppress macrophage activation in vitro and alleviate chemically induced colitis in mice. AB - Analysis of parasite-host interactions can reveal the intricacies of immunity and identify ways to modulate immunopathological reactions. We assessed the ability of a phosphate-buffered saline-soluble extract of adult Hymenolepis diminuta to suppress macrophage (human THP-1 cell line, murine peritoneal macrophages) activity in vitro and the impact of treating mice with this extract on colitis induced by dinitrobenzene sulfonic acid (DNBS). A high-molecular-mass fraction of adult H. diminuta (HdHMW) or excretory/secretory products reduced macrophage activation: lipopolysaccharide (LPS)-induced interleukin-1beta (IL-1beta), IL-6, and tumor necrosis factor alpha (TNF-alpha) and poly(I:C)-induced TNF-alpha and IL-6 were suppressed by HdHMW. The active component in the HdHMW extract was minimally sensitive to boiling and trypsin digestion, whereas the use of sodium metaperiodate, as a general deglycosylation strategy, indicated that the immunosuppressive effect of HdHMW was at least partially dependent on a glycan: treating the HdHMW with neuraminidase and alpha-mannosidase failed to inhibit its blockade of LPS-induced TNF-alpha production by THP-1 macrophages. Mice treated with DNBS developed colitis, as typified by wasting, shortening of the colon, macroscopic and microscopic tissue damage, and an inflammatory infiltrate. Mice cotreated with HdHMW (three intraperitoneal injections) displayed significantly less inflammatory disease, and this was accompanied by reduced TNF-alpha production and increased IL-10 and IL-4 production by mitogen-stimulated spleen cells. However, cotreatment of mice with neutralizing anti-IL-10 antibodies had only a minor impact on the anticolitic effect of the HdHMW. We speculate that purification of the immunosuppressive factor(s) from H. diminuta has the potential to lead to the development of novel immunomodulatory drugs to treat inflammatory disease. PMID- 20028814 TI - Comparison of anidulafungin MICs determined by the clinical and laboratory standards institute broth microdilution method (M27-A3 document) and Etest for Candida species isolates. AB - Anidulafungin Etest and CLSI MICs were compared for 143 Candida sp. isolates to assess essential (within 2 log(2) dilutions) and categorical agreements (according to three susceptibility breakpoints). Based on agreement percentages, our data indicated that Etest is not suitable to test anidulafungin against Candida parapsilosis and C. guilliermondii (54.4 to 82.4% essential and categorical agreements) but is more suitable for C. albicans, C. glabrata, C. krusei, and C. tropicalis (87.9 to 100% categorical agreement). PMID- 20028813 TI - Meningococcal porin PorB prevents cellular apoptosis in a toll-like receptor 2- and NF-kappaB-independent manner. AB - Meningococcal porin PorB is an inhibitor of apoptosis induced via the intrinsic pathway in various cell types. This effect is attributed to prevention of mitochondrial depolarization and of subsequent release of proapoptotic mitochondrial factors. To determine whether apoptosis is globally inhibited by PorB, we compared the intrinsic and extrinsic pathways in HeLa cells. Interestingly, PorB does not prevent extrinsic apoptosis induced by tumor necrosis factor alpha plus cycloheximide, suggesting a unique mitochondrial pathway specificity. Several intracellular factors regulated by NF-kappaB, including members of the Bcl-2 family and of the inhibitor of apoptosis (IAP) family, play major roles in controlling apoptosis, and some of them are thought to contribute to the antiapoptotic effect of the gonococcal porin, PIB. However, most of the members of the Bcl-2 family and the IAP family are not induced by meningococcal PorB in HeLa cells, with the exception of Bfl-1/A1. Interestingly, PorB does not induce NF-kappaB activation in HeLa cells, likely due to a lack of Toll-like receptor 2 (TLR2) expression in these cells. Bfl-1/A1 expression is also regulated by CBF1, a nuclear component of the Notch signaling pathway, independent of NF-kappaB activation. Since HeLa cells are protected by PorB from intrinsic apoptosis events, regardless of TLR2 and NF-kappaB expression, the possibility of a contribution of alternative signaling pathways to this effect cannot be excluded. In this paper, we describe an initial dissection of the cascade of cellular events involved in the antiapoptotic effect of PorB in the absence of TLR2. PMID- 20028815 TI - Early viral kinetics of telbivudine and entecavir: results of a 12-week randomized exploratory study with patients with HBeAg-positive chronic hepatitis B. AB - We characterized the early viral kinetic profiles of telbivudine and entecavir and the effects of these potent nucleoside analogs on hepatitis B virus (HBV) DNA and alanine aminotransferase levels in adults with hepatitis B e antigen-positive compensated chronic hepatitis B. Forty-four patients were enrolled in this open label, parallel-group, multicenter study and randomized to receive telbivudine or entecavir for 12 weeks. Reductions in hepatitis B virus DNA and alanine aminotransferase levels from baseline to weeks 2, 4, 8, and 12 were assessed. Viral kinetic parameters, including viral clearance per day, loss of infected cells per day, and efficiency of inhibition of viral production, were estimated by using a biphasic mathematical model. Statistical analyses were limited to descriptive analyses. The 2 treatment groups achieved similar reductions in HBV DNA and alanine aminotransferase levels. Mean reductions in levels of hepatitis B virus DNA at week 12 were 6.6 +/- 1.6 and 6.5 +/- 1.5 log(10) copies/ml for the telbivudine- and entecavir-treated patients, respectively. There were no significant differences between groups in values for mean viral clearance per day, mean loss of infected cells per day, or efficiency of blocking viral production. The safety profiles for both medications were favorable. During the first 12 weeks of treatment, telbivudine and entecavir demonstrated similar antiviral potencies, resulting in a rapid and profound suppression of serum hepatitis B virus DNA and reduction of alanine aminotransferase levels. No differences in the effects of these 2 agents on early viral kinetics were observed. Both medications were well tolerated. PMID- 20028816 TI - Synergistic antimicrobial activities of folic acid antagonists and nucleoside analogs. AB - The antimicrobial activities of folic acid antagonists are supposed to be antagonized by elevated extracellular thymidine concentrations in damaged host tissues. Therefore, this study was aimed at screening for nucleoside analogs that impair bacterial thymidine utilization and analyzing the combined antimicrobial activities of nucleoside analogs and folic acid antagonists in the presence of thymidine. Our screening results revealed that different nucleoside analogs, in particular halogenated derivatives of 2'-deoxyuridine, substantially impaired the bacterial utilization of extracellular thymidine in Staphylococcus aureus. Time kill methods showed that 5-iodo-2'-deoxyuridine enhanced the extent of killing of trimethoprim-sulfamethoxazole (SXT) at 24 h against S. aureus in the presence of thymidine (200 microg/liter). While SXT (40 mg/liter) alone did not kill bacteria in the presence of thymidine, its combination with the nucleoside analog at a concentration of 8 mumol/liter showed a bactericidal effect. Moreover, 5-iodo-2' deoxyuridine combined with SXT in the presence of thymidine showed a broad spectrum of activity against several Gram-positive and Gram-negative bacteria. In conclusion, these data provide evidence that the in vitro antimicrobial activity of SXT in the presence of thymidine can be significantly improved by combination with a nucleoside analog. PMID- 20028817 TI - Pharmacokinetic-pharmacodynamic modeling of alpha interferon response induced by a Toll-like 7 receptor agonist in mice. AB - Recombinant alpha interferon (IFN-alpha) is used in the treatment of hepatitis C virus (HCV)-infected patients but is not optimal in terms of efficacy or tolerability. Toll-like 7 receptor (TLR-7) agonists stimulate the innate immune system to produce, among other cytokines, IFN-alpha and are being evaluated as alternative drugs to treat HCV infection. This paper describes the application of pharmacokinetic-pharmacodynamic (PK-PD) modeling to understanding the behavior of a TLR-7 agonist [9-benzyl-8-hydroxy-2-(2-methoxyethoxy) adenine (BHMA)] in mice, using IFN-alpha as a biomarker. This is the first report of such a PK-PD model, and the conclusions may be of utility in the clinical development of TLR-7 agonists for HCV infection. PMID- 20028818 TI - Combined ramR mutation and presence of a Tn1721-associated tet(A) variant in a clinical isolate of Salmonella enterica serovar Hadar resistant to tigecycline. AB - A Salmonella enterica serovar Hadar strain resistant to tigecycline (MIC, 16 microg/ml) was isolated. Molecular characterization revealed the presence of a plasmid-borne tet(A) variant associated with Tn1721 mediating a rise of the MIC for tigecycline when transferred to Escherichia coli. Additionally, a truncating mutation in ramR was detected. Transformation with wild-type ramR but not with the mutated ramR lowered the MIC for tigecycline. Characterization of this Salmonella isolate implicates ramR in resistance to tigecycline. PMID- 20028819 TI - Acinetobacter baumannii increases tolerance to antibiotics in response to monovalent cations. AB - Acinetobacter baumannii is well adapted to the hospital environment, where infections caused by this organism are associated with significant morbidity and mortality. Genetic determinants of antimicrobial resistance have been described extensively, yet the mechanisms by which A. baumannii regulates antibiotic resistance have not been defined. We sought to identify signals encountered within the hospital setting or human host that alter the resistance phenotype of A. baumannii. In this regard, we have identified NaCl as being an important signal that induces significant tolerance to aminoglycosides, carbapenems, quinolones, and colistin upon the culturing of A. baumannii cells in physiological NaCl concentrations. Proteomic analyses of A. baumannii culture supernatants revealed the release of outer membrane proteins in high NaCl, including two porins (CarO and a 33- to 36-kDa protein) whose loss or inactivation is associated with antibiotic resistance. To determine if NaCl affected expression at the transcriptional level, the transcriptional response to NaCl was determined by microarray analyses. These analyses highlighted 18 genes encoding putative efflux transporters that are significantly upregulated in response to NaCl. Consistent with this, the effect of NaCl on the tolerance to levofloxacin and amikacin was significantly reduced upon the treatment of A. baumannii with an efflux pump inhibitor. The effect of physiological concentrations of NaCl on colistin resistance was conserved in a panel of multidrug-resistant isolates of A. baumannii, underscoring the clinical significance of these observations. Taken together, these data demonstrate that A. baumannii sets in motion a global regulatory cascade in response to physiological NaCl concentrations, resulting in broad-spectrum tolerance to antibiotics. PMID- 20028820 TI - Mechanism of hepatitis C virus RNA polymerase inhibition with dihydroxypyrimidines. AB - We studied the biochemical mechanisms associated with inhibition and resistance to a 4,5-dihydroxypyrimidine carboxylate that inhibits the hepatitis C virus (HCV) RNA-dependent RNA polymerase NS5B. On the basis of the structure of the pharmacophore, it has been suggested that these compounds may act as pyrophosphate (PP(i)) mimics. We monitored nucleotide incorporation events during the elongation phase and showed that the polymerase activity of wild-type NS5B was inhibited by the dihydroxypyrimidine at a 50% inhibitory concentration (IC(50)) of 0.73 muM. Enzymes with the G152E or P156L mutation, either of which confers resistance to this compound, showed four- to fivefold increases in IC(50)s. The inhibitor was competitive with respect to nucleotide incorporation. It was likewise effective at preventing the PP(i)-mediated excision of an incorporated chain terminator in a competitive fashion. In the absence of the dihydroxypyrimidine, the reaction was not significantly affected by the G152E or P156L mutation. These data suggest that the resistance associated with these two mutations is unlikely due to an altered interaction with the pyrophosphate mimicking domain of the compound but, rather, is due to altered interactions with its specificity domain at a region distant from the active site. Together, our findings provide strong experimental evidence that supports the notion that the members of this class of compounds can act as PP(i) mimics that have the potential to mechanistically complement established nucleoside and nonnucleoside analogue inhibitors. PMID- 20028821 TI - Antimalarial asexual stage-specific and gametocytocidal activities of HIV protease inhibitors. AB - The stage-specific antimalarial activities of a panel of antiretroviral protease inhibitors (PIs), including two nonpeptidic PIs (tipranavir and darunavir), were tested in vitro against Plasmodium falciparum. While darunavir demonstrated limited antimalarial activity (effective concentration [EC(50)], >50 microM), tipranavir was active at clinically relevant concentrations (EC(50), 12 to 21 microM). Saquinavir, lopinavir, and tipranavir preferentially inhibited the growth of mature asexual-stage parasites (24 h postinvasion). While all of the PIs tested inhibited gametocytogenesis, tipranavir was the only one to exhibit gametocytocidal activity. PMID- 20028822 TI - Exploiting the drug-activating properties of a novel trypanosomal nitroreductase. AB - Nitroheterocyclic prodrugs have been used to treat trypanosomal diseases for more than 40 years. Recently, the key step involved in the activation of these compounds has been elucidated and shown to be catalyzed by a type I nitroreductase (NTR). This class of enzyme is normally associated with bacteria and is absent from most eukaryotes, with trypanosomes being a major exception. Here we exploit this difference by evaluating the trypanocidal activity of a library of nitrobenzylphosphoramide mustards against bloodstream-form Trypanosoma brucei parasites. Biochemical screening against the purified enzyme revealed that a subset of halogenated nitroaromatic compounds were effective substrates for T. brucei NTR (TbNTR), having apparent K(cat)/K(m) values approximately 100 times greater than nifurtimox. When tested against T. brucei, cytotoxicity mirrored enzyme activity, with 50% inhibitory concentrations of the most potent substrates being less than 10 nM. T. brucei NTR plays a key role in parasite killing: heterozygous lines displayed resistance to the compounds, while parasites overexpressing the enzyme showed hypersensitivity. We also evaluated the cytotoxicities of substrates with the highest trypanocidal activities by using mammalian THP-1 cells. The relative toxicities of these newly identified compounds were much lower than that of nifurtimox. We conclude that halogenated nitrobenzylphosphoramide mustards represent a novel class of antitrypanosomal agents, and their efficacy validates the strategy of specifically targeting NTR activity to develop new therapeutics. PMID- 20028823 TI - Spread of a chromosomal cefixime-resistant penA gene among different Neisseria gonorrhoeae lineages. AB - In Neisseria gonorrhoeae, the mosaic type of penA, which encodes penicillin binding protein 2 (PBP 2), is associated with reduced susceptibility to oral cephalosporins. To investigate the relatedness of N. gonorrhoeae clinical isolates with reduced susceptibility, we sequenced the penA genes of 32 isolates. Five different amino acid sequence types of PBP 2 were identified, but all seemed to be derivatives of pattern X of PBP 2 (PBP 2-X). However, multilocus sequence typing of the isolates showed that the isolates belonged to six different sequence types. As PBP 2-X was identified in three different sequence types, horizontal transfer of the penA allele encoding PBP2-X was suggested. We demonstrated that the penA gene could be transferred from an isolate with reduced susceptibility to a sensitive isolate by natural transformation. Comparison of the sequence of the penA-flanking regions of 12 transformants with those of the donor and the recipient suggested that at least a 4-kb DNA segment, including the penA gene, was transferred. During horizontal transfer, some of the penA alleles also acquired variations due to point mutations and genetic exchange within the allele. Our results provide evidence that the capacity for natural transformation in N. gonorrhoeae plays a role in the spread of chromosomal antibiotic resistance genes and the generation of diversity in such genes. PMID- 20028824 TI - Elucidation of the pharmacokinetic/pharmacodynamic determinant of colistin activity against Pseudomonas aeruginosa in murine thigh and lung infection models. AB - Colistin is increasingly used as last-line therapy against Gram-negative pathogens. The pharmacokinetic (PK)/pharmacodynamic (PD) index that best correlates with the efficacy of colistin remains undefined. The activity of colistin against three strains of Pseudomonas aeruginosa was studied in neutropenic mouse thigh and lung infection models. The PKs of unbound colistin were determined from single-dose PK studies together with extensive plasma protein binding analyses. Dose-fractionation studies were conducted over 24 h with a dose range of 5 to 160 mg/kg of body weight/day. The bacterial burden in the thigh or lung was measured at 24 h after the initiation of treatment. Relationships between antibacterial effect and measures of exposure to unbound (f) colistin (area under the concentration-time curve [fAUC/MIC], maximum concentration of drug in plasma [fC(max)]/MIC, and the time that the concentration in plasma is greater than the MIC [fT > MIC]) were examined by using an inhibitory sigmoid maximum-effect model. Nonlinearity in the PKs of colistin, including its plasma protein binding, was observed. The PK/PD index that correlated best with its efficacy was fAUC/MIC in both the thigh infection model (R(2) = 87%) and the lung infection model (R(2) = 89%). The fAUC/MIC targets required to achieve 1-log and 2-log kill against the three strains were 15.6 to 22.8 and 27.6 to 36.1, respectively, in the thigh infection model, while the corresponding values were 12.2 to 16.7 and 36.9 to 45.9 in the lung infection model. The findings of this in vivo study indicate the importance of achieving adequate time-averaged exposure to colistin. The results will facilitate efforts to define the more rational design of dosage regimens for humans. PMID- 20028825 TI - Probing the antimalarial mechanism of artemisinin and OZ277 (arterolane) with nonperoxidic isosteres and nitroxyl radicals. AB - Peroxidic antimalarials such as the semisynthetic artemisinins are critically important in the treatment of drug-resistant malaria. Nevertheless, their peroxide bond-dependent mode of action is still not well understood. Using combination experiments with cultured Plasmodium falciparum cells, we investigated the interactions of the nitroxide radical spin trap, 2,2,6,6 tetramethyl-1-piperidinyloxy (TEMPO), and four of its analogs with artemisinin and the ozonide drug development candidate OZ277. The antagonism observed for combinations of artemisinin or OZ277 with the TEMPO analogs supports the hypothesis that the formation of carbon-centered radicals is critical for the activity of these two antimalarial peroxides. The TEMPO analogs showed a trend toward greater antagonism with artemisinin than they did with OZ277, an observation that can be explained by the greater tendency of artemisinin-derived carbon-centered radicals to undergo internal self-quenching reactions, resulting in a lower proportion of radicals available for subsequent chemical reactions such as the alkylation of heme and parasite proteins. In a further mechanistic experiment, we tested both artemisinin and OZ277 in combination with their nonperoxidic analogs. The latter had no effect on the antimalarial activities of the former. These data indicate that the antimalarial properties of peroxides do not derive from reversible interactions with parasite targets. PMID- 20028826 TI - Detection of molecular markers of drug resistance in 2009 pandemic influenza A (H1N1) viruses by pyrosequencing. AB - The M2 blockers amantadine and rimantadine and the neuraminidase (NA) inhibitors (NAIs) oseltamivir and zanamivir are approved by the FDA for use for the control of influenza A virus infections. The 2009 pandemic influenza A (H1N1) viruses (H1N1pdm) are reassortants that acquired M and NA gene segments from a Eurasian adamantane-resistant swine influenza virus. NAI resistance in the H1N1pdm viruses has been rare, and its occurrence is mainly limited to oseltamivir-exposed patients. The pyrosequencing assay has been proven to be a useful tool in surveillance for drug resistance in seasonal influenza A viruses. We provide a protocol which allows the detection of adamantane resistance markers as well as the I43T change, which is unique to the H1N1pdm M2 protein. The protocol also allows the detection of changes at residues V116, I117, E119, Q136, K150, D151, D199, I223, H275, and N295 in the NA, known to alter NAI drug susceptibility. We report on the detection of the first cases of the oseltamivir resistance conferring mutation H275Y and the I223V change in viruses from the United States using the approach described in this study. Moreover, the assay permits the quick identification of the major NA group (V106/N248, I106/D248, or I106/N248) to which a pandemic virus belongs. Pyrosequencing is well suited for the detection of drug resistance markers and signature mutations in the M and NA gene segments of the pandemic H1N1 influenza viruses. PMID- 20028827 TI - Effects of renal volume and single-kidney glomerular filtration rate on renal functional outcome in atherosclerotic renal artery stenosis. AB - BACKGROUND: Renal functional outcome is unpredictable after revascularization of high-grade atherosclerotic renal artery stenosis (RAS). 'Hibernating' parenchyma describes acute parenchymal injury where renal dysfunction is potentially reversible with treatment of the stenosis. We analysed renal parenchymal volume (PV) and single-kidney glomerular filtration rate (SK-GFR) characteristics to identify kidneys with hibernating parenchyma and hence determine renal functional outcome after revascularization. METHODS: Fifty patients with > or =50% RAS underwent baseline analyses: (i) PV using magnetic resonance imaging; (ii) radioisotopic SK-GFR. Twenty-one patients (27 kidneys) underwent renal revascularization and 29 medical therapy alone. RESULTS: Patients with revascularized kidneys manifesting high PV:SK-GFR showed improvement in global estimated GFR compared to conservatively managed counterparts at 6 months and 1 year (6 months: 6.2 +/- 2.9 versus -3.7 +/- 6.8, P = 0.038; 1 year: 3.5 +/- 3.0 versus -5.1 +/- 5.1 ml/min/1.73 m(2), P = 0.021). Twelve revascularized patients (16 kidneys) underwent repeat SK-GFR 4 months post-revascularization. Six of 16 revascularized kidneys had high baseline PV:SK-GFR and showed improved SK-GFR compared to kidneys with low or normal PV:SK-GFR (6.3 +/- 2.0 versus -0.9 +/- 4.2 ml/min, P = 0.002). CONCLUSIONS: Our data suggest that, after revascularization, GFR improvement is likely if there is a disproportionately higher baseline PV:SK GFR in the RAS kidney. Analysing these parameters can potentially identify these 'hibernating' kidneys and aid determination of renal functional outcome in RAS. PMID- 20028828 TI - The Rowett rat strain is resistant to renal fibrosis. AB - BACKGROUND: Genetic susceptibility to renal fibrosis may determine the individual rate of progression to renal failure. We aimed to study the progression in Rowett (RO) rats, a strain we found resistant to subtotal nephrectomy (SNX), comparing to Sprague-Dawley (SD) rats, a strain with established sensitivity in a radical ablation/infarction and diet-induced SNX model. METHODS: Eight-week-old male RO (RO-SNX) and SD (SD-SNX, n = 5/group) rats underwent SNX and were kept on high protein and salt diet. Kidney function was monitored and the kidneys were evaluated by histology and immunohistochemistry 5 weeks after SNX. RESULTS: RO SNX rats had only mild proteinuria and less glomerulosclerosis, accompanied by less fibronectin and TGF-beta staining as compared to SD-SNX rats. Glomerular nitrotyrosine staining was less intense in RO-SNX vs SD-SNX, accompanied by less podocyte damage as demonstrated by desmin staining. CONCLUSION: Our results demonstrate the importance of podocyte damage in glomerulosclerosis and that Rowett rats are protected from renal fibrosis. To our knowledge, this is the first strain of rats with unknown genetic resistance, which makes the strain attractive for studying the genetic background of renal fibrosis. PMID- 20028829 TI - High-flux or low-flux dialysis: a position statement following publication of the Membrane Permeability Outcome study. PMID- 20028830 TI - Conditioned pharmacotherapeutic effects: a preliminary study. AB - OBJECTIVE: To test the hypothesize that psoriasis patients treated under a partial schedule of pharmacologic (corticosteroid) reinforcement would show less severe symptoms and relapse than those given the same amount of drug under standard conditions. Behavioral conditioning as an inherent component of many pharmacotherapeutic protocols has never been examined. METHODS: A double-blind, simple randomization intervention was conducted with 46 patients from California and New York. Initially, lesions were treated with 0.1% acetonide triamcinolone under standard treatment conditions. Thereafter, a Standard Therapy group continued on continuous reinforcement (active drug every treatment) with 100% of the initial dose; Partial Reinforcement patients received a full dose 25% to 50% of the time and placebo medication other times; Dose Control patients received continuous reinforcement with 25% to 50% of the initial dose. RESULTS: Severity of disease scores in California neither supported nor refuted the hypothesis. In New York, where there was no difference between Partial Reinforcement and Dose Control groups at baseline, partial reinforcement effected a greater reduction in lesion severity than Dose Control conditions and did not differ from Standard Therapy patients receiving two to four times more drug. For the entire population, the frequency of relapse under partial reinforcement (26.7%) was lower than in Dose Control patients (61.5%) and did not differ from full-dose treatment (22.2%). CONCLUSIONS: A partial schedule of pharmacotherapeutic reinforcement could maintain psoriasis patients with a cumulative amount of corticosteroid that was relatively ineffective when administered under standard treatment conditions. Conceivably, corticosteroid administration only one quarter or half as frequently as currently prescribed is sufficient to treat psoriasis. We posit, however, that these preliminary observations implicate conditioning processes in-and for the design of-regimens of pharmacotherapy. PMID- 20028831 TI - Heart rate recovery after cognitive challenge is preserved with age. AB - OBJECTIVE: To investigate the effect of age on heart rate recovery (HRR) from cognitive challenge. BACKGROUND: Aging is an independent risk factor for the development of cardiovascular disease. HRR from exercise is an established predictor of cardiac morbidity and mortality, and evidence suggests that HRR from cognitive challenge is predictive of cardiac morbidity as well. Aging is associated with delayed HRR from exercise stress, but little is known about the effect of aging on HRR from psychological stress. We tested the hypothesis that age would be related to delayed HRR from psychological stress. METHODS: HRR post exposure to cognitive challenge (mental arithmetic and Stroop) was investigated in a sample of 436 participants aged 35 to 84 years in MIDUS II, a national study of health and well-being. HRR was measured as 1) the amount of change from the stress level; 2) time to recover; and 3) the area under the curve. The analyses were controlled for medical comorbidities and medications that influence HR, such as body mass index, smoking, sex, menopausal status, and amount of physical activity/exercise. RESULTS: There was no effect for age on HRR as evaluated by all three recovery assessment methods. CONCLUSIONS: Contrary to expectation and in contrast to findings concerning HRR from exercise, HRR from cognitive challenge was preserved with age. These findings require further inquiry into differential mechanism(s) underlying HRR from psychological versus exercise stress, including any role for improved emotion regulation with greater age. PMID- 20028832 TI - Rationale and design of a randomized clinical trial comparing stress reduction treatment to usual cardiac care: the Reducing Vulnerability to Implantable Cardioverter Defibrillator Shock-Treated Ventricular Arrhythmias (RISTA) trial. AB - OBJECTIVE: To present the design of a multicenter, randomized trial testing the effects of stress reduction treatment (SRT) on the prevalence of shock-treated ventricular arrhythmias among patients with an implantable cardioverter defibrillator (ICD). Significant adjustment problems secondary to ICD shock can increase the likelihood of arrhythmias requiring shock for termination. Whether SRT can reduce arrhythmias requiring shock for termination in patients with ICDs has not been tested in clinical trials. METHODS: New ICD recipients and previous recipients who have received an appropriate therapeutic shock in the last 6 months (n = 304) will be enrolled and randomized to either SRT or usual cardiac care. Participants complete a psychosocial questionnaire and undergo laboratory mental stress testing and 24-hour Holter monitoring with diary at study entry and approximately 4 months later. Follow-ups are completed at 6, 12, and 24 months post randomization to assess occurrence of ICD shock for ventricular arrhythmias (primary outcome), antitachycardia pacing events, medication changes, hospitalizations, deaths, and quality of life. RESULTS: Log-rank test and Cox proportional hazards model will be used to test the effects of SRT on time to first shock-treated ventricular arrhythmia, with exploratory analyses testing the effects on overall frequency of ventricular arrhythmia. Secondary analyses will test the effects of SRT on laboratory stress-induced and 24-hour arrhythmogenic electrophysiological indices from pre to post treatment, and both quality of life and measures of anger across the 2 years of the study. CONCLUSIONS: The Reducing Vulnerability to ICD Shock-Treated Ventricular Arrhythmias (RISTA) Trial is the first large-scale, randomized, clinical trial designed to evaluate the effect of SRT on the prevalence of shock-treated arrhythmias among patients with an ICD. Results may demonstrate a treatment that can reduce vulnerability to arrhythmia provoked shock and improve quality of life. PMID- 20028833 TI - Cardiac autonomic control and treatment of hostility: a randomized controlled trial. AB - OBJECTIVE: To test whether reduction in hostility increases autonomic regulation of the heart. METHODS: In this randomized controlled trial, participants were 158 healthy adults, aged 20 years to 45 years, who were 1 standard deviation (SD) above national norms on the Cook-Medley Hostility and the Spielberger Trait Anger Indices. Participants also were interviewed, using the Interpersonal Hostility Assessment Technique (IHAT). They were randomly assigned to a 12-week cognitive behavior therapy program for hostility reduction or a wait-list control condition. The main outcome measure was cardiac autonomic modulation, measured as RR interval variability (RRV) derived from 24-electrocardiographic recordings. RESULTS: In a multivariate analysis of variance assessing psychological outcomes of hostility, anger, and IHAT scores, there was a significant treatment effect with an average reduction across the three outcomes that was approximately 0.7 SD (ES = 0.685, SE = 0.184, p < .001) greater for the intervention group than for the control group. In contrast, the change in heart rate was -0.14 beat/min (95% Confidence Interval [CI] = -2.43, 2.14) in treatment participants and -1.36 beat/min (95% CI = -3.28, 0.61) in wait-list participants. High-frequency RRV, an index of cardiac parasympathetic modulation, increased by 0.07 ln ms(2) (95% CI = -0.10, 0.24) for participants in the treatment condition and decreased by 0.04 ln ms(2) (95% CI = -0.18, 0.10) for participants in the wait-list condition. These differences were not significant. The findings for other indices of RRV were similar. CONCLUSIONS: Reduction of hostility and anger was not accompanied by increases in cardiac autonomic modulation. These findings raise questions about the status of disordered autonomic nervous system regulation of the heart as a pathophysiological mechanism underlying the hostility-heart disease relationship and about whether hostility itself is a mechanism or merely a marker of elevated risk of heart disease. PMID- 20028836 TI - What's new in hypertension 2009? PMID- 20028838 TI - Heat shock protein cognate 70-4 and an E3 ubiquitin ligase, CHIP, mediate plastid destined precursor degradation through the ubiquitin-26S proteasome system in Arabidopsis. AB - Plastid-targeted proteins pass through the cytosol as unfolded precursors. If proteins accumulate in the cytosol, they can form nonspecific aggregates that cause severe cellular damage. Here, we demonstrate that high levels of plastid precursors are degraded through the ubiquitin-proteasome system (UPS) in Arabidopsis thaliana cells. The cytosolic heat shock protein cognate 70-4 (Hsc70 4) and E3 ligase carboxy terminus of Hsc70-interacting protein (CHIP) were highly induced in plastid protein import2 plants, which had a T-DNA insertion at Toc159 and showed an albino phenotype and a severe defect in protein import into chloroplasts. Hsc70-4 and CHIP together mediated plastid precursor degradation when import-defective chloroplast-targeted reporter proteins were transiently expressed in protoplasts. Hsc70-4 recognized specific sequence motifs in transit peptides and thereby led to precursor degradation through the UPS. CHIP, which interacted with Hsc70-4, functioned as an E3 ligase in the Hsc70-4-mediated protein degradation. The physiological role of Hsc70-4 was confirmed by analyzing Hsc70-4 RNA interference plants in an hsc70-1 mutant background. Plants with lower Hsc70 levels exhibited abnormal embryogenesis, resulting in defective seedlings that displayed high levels of reactive oxygen species and monoubiquitinated Lhcb4 precursors. We propose that Hsc70-4 and CHIP mediate plastid-destined precursor degradation to prevent cytosolic precursor accumulation and thereby play a critical role in embryogenesis. PMID- 20028839 TI - The small subunit of snapdragon geranyl diphosphate synthase modifies the chain length specificity of tobacco geranylgeranyl diphosphate synthase in planta. AB - Geranyl diphosphate (GPP), the precursor of many monoterpene end products, is synthesized in plastids by a condensation of dimethylallyl diphosphate and isopentenyl diphosphate (IPP) in a reaction catalyzed by homodimeric or heterodimeric GPP synthase (GPPS). In the heterodimeric enzymes, a noncatalytic small subunit (GPPS.SSU) determines the product specificity of the catalytic large subunit, which may be either an active geranylgeranyl diphosphate synthase (GGPPS) or an inactive GGPPS-like protein. Here, we show that expression of snapdragon (Antirrhinum majus) GPPS.SSU in tobacco (Nicotiana tabacum) plants increased the total GPPS activity and monoterpene emission from leaves and flowers, indicating that the introduced catalytically inactive GPPS.SSU found endogenous large subunit partner(s) and formed an active snapdragon/tobacco GPPS in planta. Bimolecular fluorescence complementation and in vitro enzyme analysis of individual and hybrid proteins revealed that two of four GGPPS-like candidates from tobacco EST databases encode bona fide GGPPS that can interact with snapdragon GPPS.SSU and form a functional GPPS enzyme in plastids. The formation of chimeric GPPS in transgenic plants also resulted in leaf chlorosis, increased light sensitivity, and dwarfism due to decreased levels of chlorophylls, carotenoids, and gibberellins. In addition, these transgenic plants had reduced levels of sesquiterpene emission, suggesting that the export of isoprenoid intermediates from the plastids into the cytosol was decreased. These results provide genetic evidence that GPPS.SSU modifies the chain length specificity of phylogenetically distant GGPPS and can modulate IPP flux distribution between GPP and GGPP synthesis in planta. PMID- 20028840 TI - Phosphorylation of photosystem II controls functional macroscopic folding of photosynthetic membranes in Arabidopsis. AB - Photosynthetic thylakoid membranes in plants contain highly folded membrane layers enriched in photosystem II, which uses light energy to oxidize water and produce oxygen. The sunlight also causes quantitative phosphorylation of major photosystem II proteins. Analysis of the Arabidopsis thaliana stn7xstn8 double mutant deficient in thylakoid protein kinases STN7 and STN8 revealed light independent phosphorylation of PsbH protein and greatly reduced N-terminal phosphorylation of D2 protein. The stn7xstn8 and stn8 mutants deficient in light induced phosphorylation of photosystem II had increased thylakoid membrane folding compared with wild-type and stn7 plants. Significant enhancement in the size of stacked thylakoid membranes in stn7xstn8 and stn8 accelerated gravity driven sedimentation of isolated thylakoids and was observed directly in plant leaves by transmission electron microscopy. Increased membrane folding, caused by the loss of light-induced protein phosphorylation, obstructed lateral migration of the photosystem II reaction center protein D1 and of processing protease FtsH between the stacked and unstacked membrane domains, suppressing turnover of damaged D1 in the leaves exposed to high light. These findings show that the high level of photosystem II phosphorylation in plants is required for adjustment of macroscopic folding of large photosynthetic membranes modulating lateral mobility of membrane proteins and sustained photosynthetic activity. PMID- 20028841 TI - Quantitative proteomics of the tonoplast reveals a role for glycolytic enzymes in salt tolerance. AB - To examine the role of the tonoplast in plant salt tolerance and identify proteins involved in the regulation of transporters for vacuolar Na(+) sequestration, we exploited a targeted quantitative proteomics approach. Two dimensional differential in-gel electrophoresis analysis of free flow zonal electrophoresis separated tonoplast fractions from control, and salt-treated Mesembryanthemum crystallinum plants revealed the membrane association of glycolytic enzymes aldolase and enolase, along with subunits of the vacuolar H(+) ATPase V-ATPase. Protein blot analysis confirmed coordinated salt regulation of these proteins, and chaotrope treatment indicated a strong tonoplast association. Reciprocal coimmunoprecipitation studies revealed that the glycolytic enzymes interacted with the V-ATPase subunit B VHA-B, and aldolase was shown to stimulate V-ATPase activity in vitro by increasing the affinity for ATP. To investigate a physiological role for this association, the Arabidopsis thaliana cytoplasmic enolase mutant, los2, was characterized. These plants were salt sensitive, and there was a specific reduction in enolase abundance in the tonoplast from salt treated plants. Moreover, tonoplast isolated from mutant plants showed an impaired ability for aldolase stimulation of V-ATPase hydrolytic activity. The association of glycolytic proteins with the tonoplast may not only channel ATP to the V-ATPase, but also directly upregulate H(+)-pump activity. PMID- 20028842 TI - Orthologs of the class A4 heat shock transcription factor HsfA4a confer cadmium tolerance in wheat and rice. AB - Cadmium (Cd) is a widespread soil pollutant; thus, the underlying molecular controls of plant Cd tolerance are of substantial interest. A screen for wheat (Triticum aestivum) genes that confer Cd tolerance to a Cd hypersensitive yeast strain identified Heat shock transcription factor A4a (HsfA4a). Ta HsfA4a is most similar to the class A4 Hsfs from monocots. The most closely related rice (Oryza sativa) homolog, Os HsfA4a, conferred Cd tolerance in yeast, as did Ta HsfA4a, but the second most closely related rice homolog, Os HsfA4d, did not. Cd tolerance was enhanced in rice plants expressing Ta HsfA4a and decreased in rice plants with knocked-down expression of Os HsfA4a. An analysis of the functional domain using chimeric proteins constructed from Ta HsfA4a and Os HsfA4d revealed that the DNA binding domain (DBD) of HsfA4a is critical for Cd tolerance, and within the DBD, Ala-31 and Leu-42 are important for Cd tolerance. Moreover, Ta HsfA4a-mediated Cd resistance in yeast requires metallothionein (MT). In the roots of wheat and rice, Cd stress caused increases in HsfA4a expression, together the MT genes. Our findings thus suggest that HsfA4a of wheat and rice confers Cd tolerance by upregulating MT gene expression in planta. PMID- 20028843 TI - Drosophila brain development: closing the gap between a macroarchitectural and microarchitectural approach. AB - Neurobiologists address neural structure, development, and function at the level of "macrocircuits" (how different brain compartments are interconnected; what overall pattern of activity they produce) and at the level of "microcircuits" (how connectivity and physiology of individual neurons and their processes within a compartment determine the functional output of this compartment). Work in our lab aims at reconstructing the developing Drosophila brain at both levels. Macrocircuits can be approached conveniently by reconstructing the pattern of brain lineages, which form groups of neurons whose projections form cohesive fascicles interconnecting the compartments of the larval and adult brain. The reconstruction of microcircuits requires serial section electron microscopy, due to the small size of terminal neuronal processes and their synaptic contacts. Because of the amount of labor that traditionally comes with this approach, very little is known about microcircuitry in brains across the animal kingdom. Many of the problems of serial electron microscopy reconstruction are now solvable with digital image recording and specialized software for both image acquisition and postprocessing. In this chapter, we introduce our efforts to reconstruct the small Drosophila larval brain and discuss our results in light of the published data on neuropile ultrastructure in other animal taxa. PMID- 20028844 TI - snaR genes: recent descendants of Alu involved in the evolution of chorionic gonadotropins. AB - We identified a novel family of human noncoding RNAs by in vivo cross-linking to the nuclear factor 90 (NF90) protein. These small NF90-associated RNAs (snaRs) are transcribed by RNA polymerase III and display restricted tissue distribution, with high expression in testis and discrete areas of the brain. The most abundant human transcript, snaR-A, interacts with the cell's transcription and translation systems. snaR genes have evolved in African Great Apes (human, chimpanzee, and gorilla) and some are unique to humans. We traced their ancestry to the Alu SINE (short interspersed nucleotide element) family, via two hitherto unreported sets of short genetic elements termed ASR (Alu/snaR-related) and CAS (Catarrhine ancestor of snaR). This derivation entails a series of internal deletions followed by expansions. The evolution of these genes coincides with major primate speciation events: ASR elements are found in all monkeys and apes, whereas CAS elements are limited to Old World monkeys and apes. In contrast to ASR and CAS elements, which are retrotransposons, human snaR genes are predominantly located in three clusters on chromosome 19 and have been duplicated as part of a larger genetic element. Insertion of the element containing snaR-G into a gene encoding a chorionic gonadotropin beta subunit generated new hormone genes in African Great Apes. PMID- 20028845 TI - The religious essence of intelligent design. AB - Despite the protestations of its proponents, "intelligent design theory" (ID) is not science but creationism, making it in its essence a supernaturalist religious belief. This fact has been established conclusively for the legal record in Kitzmiller et al. v. Dover Area School District (2005) and for the public by a decade of scholarship, much of which helped to secure the Kitzmiller plaintiffs' victory in this first legal case involving ID. Moreover, ID is not merely a religious belief but, more specifically, a narrow form of sectarian Christianity, as specified by its own proponents. The nature of ID as a creationist, therefore religious, movement has been revealed not only by its critics, but also, most importantly, by its proponents. The explication of ID by its critics as creationism, and therefore religion, reflects the way the movement views itself. PMID- 20028848 TI - Revision ulnar collateral ligament reconstruction using a suspension button fixation technique. AB - BACKGROUND: Revision ulnar collateral ligament reconstruction remains a challenging problem. The objective of this study was to biomechanically evaluate an ulnar collateral ligament reconstruction technique using a suspension button fixation technique that can be used even in the case of ulnar cortical bone loss. HYPOTHESIS: An ulnar suspension fixation technique for ulnar collateral ligament reconstruction can restore elbow kinematics and demonstrate failure strength comparable to that of currently available techniques. STUDY DESIGN: Controlled laboratory study. METHODS: Nine pairs of cadaveric elbows were dissected free of soft tissue and potted. After simulating ulnar cortical bone loss, ulnar collateral ligament reconstruction was performed in 1 elbow of each pair using palmaris longus autograft and a 30-mm RetroButton suspended from the far (lateralmost) ulnar cortex. A docking technique was used for humeral fixation of the graft. Elbow valgus angle was quantified using a Microscribe 3DLX digitizer at multiple elbow flexion angles. Valgus angle was measured with the ulnar collateral ligament intact, transected, and reconstructed. In addition, load-to failure testing was performed in 1 elbow of each pair. RESULTS: Release of the ulnar collateral ligament caused a significant increase in valgus angle at each flexion angle tested (P < .002). Reconstructed elbows demonstrated no significant differences in valgus angle from the intact elbow at all flexion angles tested. Load-to-failure tests showed that reconstructed elbows had an ultimate torque (10.3 + or - 5.7 N x m) significantly less than intact elbows (26.4 + or - 10.6 N x m) (P = .001). CONCLUSION: Ulnar collateral ligament reconstruction using a suspension button fixation technique reliably restored elbow kinematics to the intact state. Load-to-failure testing demonstrated comparable fixation strength to several historic controls of primary reconstruction techniques despite the simulated ulnar cortical bone loss. CLINICAL RELEVANCE: Ulnar collateral ligament reconstruction using a suspension button fixation technique can be considered in the case of ulnar cortical bone loss in a primary or revision setting. PMID- 20028846 TI - Genetic regulation of mammalian diversity. AB - Mammals have evolved a variety of morphological adaptations that have allowed them to compete in their natural environments. The developmental genetic basis of this morphological diversity remains largely unknown. Bats are mammals that have the unique ability of powered flight. We have examined the molecular embryology of bats and investigated the developmental genetic basis for their highly derived limbs used for flight. Initially, we developed an embryo staging system for a model chiropteran, Carollia perspicillata, the short-tailed fruit bat that has subsequently been used for staging other bat species. Expression studies focusing on genes that regulate limb development indicate that there are similarities and differences between bats and mice. To determine the consequences of these expression differences, we have conducted an enhancer switch assay by gene targeting in mouse embryonic stem cells to create mice whose genes are regulated by bat sequences. Our studies indicate that cis-regulatory elements contribute to the morphological differences that have evolved among mammalian species. PMID- 20028847 TI - Prospective evaluation of arthroscopic bankart repairs for anterior instability. AB - BACKGROUND: Arthroscopic treatment has evolved to become the primary surgical option in the management of anterior shoulder instability as studies show comparable outcomes between open and arthroscopic techniques. OBJECTIVE: To evaluate prospectively the results of our institutional database for arthroscopic Bankart repairs at a minimum 2-year follow-up for patients with anterior instability treated with suture anchors. STUDY DESIGN: Case series; Level of evidence, 4. METHODS: Eighty-three consecutive patients underwent arthroscopic Bankart repair with suture anchors. The mean age at the time of surgery was 33 years (range, 15-55 years). At an average follow-up of 33 months (range, 24-49 months), 73 patients (61 males, 12 females) were assessed with outcomes scores including the American Shoulder and Elbow Surgeons, L'Insalata, and visual analog scores. The rate of recurrent instability, range of motion, and risk factors for postoperative recurrence were evaluated. RESULTS: Thirteen patients (18%) suffered a recurrence after surgery. Seven patients (10%) had a subsequent dislocation and 6 (8%) a subluxation event or apprehension. Six of the 13 had a traumatic event that resulted in recurrent episodes of instability. Revision surgery was needed for 2 patients (3%) for instability and 2 for postoperative shoulder stiffness. On average there was no significant loss of external rotation postoperatively (average, 71 degrees pre- and postoperatively). The American Shoulder and Elbow Surgeons and L'Insalata scores improved from 75.4 to 94.9 and 66.5 to 90.9, respectively (P <.0001). The visual analog score improved from 2.4 to 0.4 (P <.001). Patient age under 25, ligamentous laxity, and the presence of a large (>250 mm(3)) Hill-Sachs lesion were associated with recurrence (P <.05). Patients under age 20 had a 37.5% recurrence rate. CONCLUSION: In the arthroscopic treatment of anterior instability, identification of risk factors for recurrence allows for appropriate patient counseling and consideration of open stabilization. In our series, patients under age 25, with ligamentous laxity, and with a large (>250 mm(3)) Hill-Sachs lesion were at the greatest risk of recurrence. PMID- 20028849 TI - Effects of exercise on muscle glycogen synthesis signalling and enzyme activities in pigs carrying the PRKAG3 mutation. AB - The dominant RN mutation in pigs results in excessive glycogen storage in skeletal muscle. The mutation is situated in the PRKAG3 gene, which encodes a muscle-specific isoform of the AMP-activated protein kinase (AMPK) gamma3 subunit. AMPK is an important regulator of carbohydrate and fat metabolism in mammalian cells. The aim of the present study was to examine the effect of exercise on glycogen synthesis signalling pathways in muscle and to study enzyme activities of importance in carbohydrate metabolism in pigs with or without the PRKAG3 mutation. Glycogen content, metabolic enzyme activities and expression or phosphorylation of signalling proteins were analysed in skeletal muscle specimens obtained at rest, after a single treadmill exercise bout and after 3 h recovery. The PRKAG3 mutation carriers had higher glycogen content, a tendency for lower expression of AMPK (P < 0.07) and higher hexokinase and phosphorylase activities, whereas citrate synthase, 3-hydroxyacyl-CoA dehydrogenase and glycogen synthase activities did not differ between genotypes. Carriers and non-carriers of the RN mutation showed a similar degradation of glycogen after exercise, whereas the rate of resynthesis was faster in the carriers. Acute exercise stimulated Akt phosphorylation on Ser(473) in both genotypes, and the effect was greater in the carriers than in the non-carriers. Acute exercise also stimulated phosphorylation of Akt substrate of 160 kDA and Glycogen synthase kinase 3 in the carriers and GSK3alpha in the non-carriers. In conclusion, the increased rate of glycogen synthesis following exercise in pigs carrying the PRKAG3 mutation correlates with an increased signalling response of Akt and its substrate, AS160, and a higher activity of hexokinase, indicating an increased glucose influx and phosphorylation of glucose, directed towards glycogen synthesis. PMID- 20028850 TI - Influence of hyperoxia on muscle metabolic responses and the power-duration relationship during severe-intensity exercise in humans: a 31P magnetic resonance spectroscopy study. AB - Severe-intensity constant-work-rate exercise results in the attainment of maximal oxygen uptake, but the muscle metabolic milieu at the limit of tolerance (T(lim)) for such exercise remains to be elucidated. We hypothesized that T(lim) during severe-intensity exercise would be associated with the attainment of consistently low values of intramuscular phosphocreatine ([PCr]) and pH, as determined using (31)P magnetic resonance spectroscopy, irrespective of the work rate and the inspired O(2) fraction. We also hypothesized that hyperoxia would increase the asymptote of the hyperbolic power-duration relationship (the critical power, CP) without altering the curvature constant (W). Seven subjects (mean +/- s.d., age 30 +/- 9 years) completed four constant-work-rate knee-extension exercise bouts to the limit of tolerance (range, 3-10 min) both in normoxia (N) and in hyperoxia (H; 70% O(2)) inside the bore of 1.5 T superconducting magnet. The [PCr] (approximately 5-10% of resting baseline) and pH (approximately 6.65) at the limit of tolerance during each of the four trials was not significantly different either in normoxia or in hyperoxia. At the same fixed work rate, the overall rate at which [PCr] fell with time was attenuated in hyperoxia (mean response time: N, 59 +/- 20 versus H, 116 +/- 46 s; P < 0.05). The CP was higher (N, 16.1 +/- 2.6 versus H, 18.0 +/- 2.3 W; P < 0.05) and the W was lower (N, 1.92 +/- 0.70 versus H, 1.48 +/- 0.31 kJ; P < 0.05) in hyperoxia compared with normoxia. These data indicate that T(lim) during severe-intensity exercise is associated with the attainment of consistently low values of muscle [PCr] and pH. The CP and W parameters of the power-duration relationship were both sensitive to the inspiration of hyperoxic gas.